PMID- 11851247 TI - Systemic lupus erythematosus and renal transplantation. Report of two cases. AB - Two cases of systemic lupus erythematosus along with terminal uremia secondary to lupus nephritis are presented. Both patients were treated with renal hemodialysis and transplantation. Certain mechanical, infectious, and immunologic problems were responsible for their death. There was no evidence of resurgence of lupus erythematosus in either patient, and the transplanted kidneys did not show evidence of lupus nephritis. Renal transplantation should be considered as a last resort in treatment of patients with terminal lupus nephritis. PMID- 11851248 TI - Penicillin sensitivity. AB - The results obtained in this study indicate that there are various patterns of hypersensitivity to penicillin. In order to detect these variations it is suggested that two antigens, penicillin and penicilloyl-polylysine, be employed to test for both serum and skin sensitivity. Absolutely accurate predictions cannot be made from these tests, but it is felt that their use will minimize untoward reactions of the anaphylactic type in patients thus screened for penicillin allergy. PMID- 11851249 TI - Sensitization to p-phenylenediamine. AB - p-Phenylenediamine (PPD) from 0.2 mg to 1.6 mg in water or in petrolatum was applied as a patch test to 501 patients with different dermatoses, asthma, hay fever, and venereal diseases. Of these patients 10.6% reacted within the first five days indicating previous hypersensitivity and 14.4% between the sixth and the 16th day indicating the initiation of hypersensitivity to PPD by the patch application (flare-up). The reactions within the first five days were not correlated with the amount of PPD in the tests, but a statistically significant correlation was found between flare-ups and amount, so that the larger the amount of PPD the higher the frequency of flare-ups. About one half of both reactions occurred in patients with eczematous dermatitis. PMID- 11851250 TI - The influence of soap on eczematous dermatitis. AB - A controlled study of the influence of soap on eczematous dermatitis of the hands (housewives' eczema) failed to show any significant influence of soap on the disease process. PMID- 11851251 TI - Discoid lupus erythematosus. Is systemic treatment necessary? AB - An evaluation of fluocinolone acetonide applied topically in the treatment of discoid lupus erythematosus is presented. Bilateral comparison of 0.025% fluocinolone acetonide cream to the cream base alone demonstrated the effectiveness of this program. In 59 patients an attempt was made to control this disease with topical fluocinolone acetonide as their only treatment. Forty three patients could be controlled in this fashion. Eleven patients responded partially, but for optimal response required a systemic supplement. Five patients did not show any improvement following topical fluocinolone acetonide applications. Noteworthy, is a group of 37 patients followed through two summer seasons. Twenty-four of these patients could be controlled with topical applications but reapplication the following summer was required. Three patients in this group cleared and remained clear. Six patients followed through two summers required a systemic supplement, while four in this group did not respond to therapy. PMID- 11851252 TI - Porphyria cutanea tarda symptomatica (PCT-S). A study of the effect of phlebotomy therapy. AB - The effect of phlebotomy therapy on porphyrin metabolism was evaluated in five patients with porphyria cutanea tarda symptomatica (PCT-S). The removal of 2,500 to 4,500 cc over a three to four and one-half month period resulted in a marked reduction in urine and fecal uroporphyrin excretion in three of the subjects. Clinical improvement accompanied the chemical response in each case. In one patient, the induced chemical and clinical remission has persisted for six months without further treatment. PMID- 11851253 TI - Cancer chemosurgery (Mohs' technique). The "chemo-check". AB - Cancer chemosurgery (Mohs' technique) was used following curettage and desiccation of 30 unusual basal cell carcinomas as a "chemo-check" to determine if a cure had been affected. Remaining tumor was found in six of these highly selected 30 cases. The remaining tumor was eradicated by chemosurgery. Using chemosurgery as a "chemo-check" can further reduce the small percentage of failures following curettage and desiccation of cutaneous carcinomas and thereby prevent difficult recurrent lesions. PMID- 11851254 TI - Topical cytotoxic therapy for cutaneous cancer and precancer. AB - The topical application of demecolcine (Colcemid, N-desacetyl methyl colchicine), N-desacetyl thiocolchicine (Thio-Colciran), and methotrexate preferably in combination, exerts a selectively destructive action on rodent (basal cell) carcinomas, solar keratoses, Bowen's disease, and keratoacanthomas, while sparing surrounding normal tissues. The former two exert their action by inhibiting mitosis in the metaphase, and the latter inhibits the commencement of mitosis by interference with the synthesis of desoxyribonucleic acid. The cosmetic results are particularly good when the ears or nose are affected, and the cure rate compares favorably with that resulting from other treatments. Although ineffective against squamous carcinomas, the fact that this form of therapy is effective for keratoacanthomas could prove a valuable additional diagnostic point between the two in doubtful cases. PMID- 11851255 TI - Dermojet histopathological artifacts. AB - The Dermojet has recently become popular as a new tool in dermatology. The injection of local anesthetic agents or of steroid preparations into the skin by pressure is a convenience for the physician and protects the patient from the psychological and physical trauma of a needle. PMID- 11851256 TI - Earlobe piercing with needles and wire. PMID- 11851257 TI - The William Allen Pusey Memorial Lecture. Dermatologic radiotherapy 1965. PMID- 11851258 TI - Percutaneous absorption of trivalent and hexavalent chromium. A comparative investigation in the guinea pig by means of 51Cr. AB - By means of an in vivo isotope technique, the so-called disappearance measurements, a comparison was made during a five-hour period between the percutaneous absorption in the guinea pig of a trivalent (chromic chloride) and a hexavalent (sodium chromate) chromium compound. Nine different concentrations of each substance were studied. We found that in the concentration interval 0.017 0.239 molar the relative absorption of the hexavalent compound was somewhat higher, but the differences were not statistically significant. On the other hand, in the higher concentrations (0.261-0.398 molar) more of the hexavalent compound was absorbed, and the differences were statistically significant. This difference in percutaneous absorption for various concentrations can perhaps account for the divergent results, in respect to the absorption of trivalent and hexavalent chromium compounds, which have recently been published. PMID- 11851259 TI - Griseofulvin-containing medium for simplified diagnosis of dermatophytosis. AB - By inoculating paired tubes of standard Sabouraud dextrose-chloramphenicol cycloheximide media, one of which contained in addition 20 micrograms/ml of griseofulvin, 86 of 88 griseofulvin-sensitive dermatophytes were recognized in 226 primary isolation cultures. Most yeast, bacteria, and mold contaminants were not selectively inhibited by the media. The method facilitates the selection of patients for oral griseofulvin therapy by those with relatively little training in mycology. PMID- 11851260 TI - Arsenical keratosis with squamous carcinoma of the skin, larynx, and lung. PMID- 11851261 TI - Methemoglobinemia associated with the ingestion of Diasone. PMID- 11851262 TI - Reiters syndrome with glomerular nephritis. PMID- 11851263 TI - Chronic disseminated lupus erythematosus and calcinosis cutis. PMID- 11851264 TI - Porphyria cutanea tarda with localized morphea. PMID- 11851265 TI - Parapsoriasis en plaques and guttate elements with secondary alopecia? (2) Alopecia probably due to a mucinous degeneration of the connective tissue component of the hair follicle? PMID- 11851266 TI - Cold urticaria. PMID- 11851268 TI - Incontinentia pigmenti. PMID- 11851267 TI - Mycetoma (Nocardia brasiliensis). PMID- 11851269 TI - Pemphigus erythematodes. PMID- 11851270 TI - Granuloma annulare (forme erythematous of Gougerot). PMID- 11851271 TI - Primary cutaneous amyloidosis. PMID- 11851272 TI - Project mouthguard: join the team to help our young athletes. AB - Project Mouthguard is a "win-win" situation for everyone involved. The athlete will be getting the best protection available for their dentition while they participate in school sports. The school will be able to prevent injuries, making sports participation safer and the job of the school's athletic trainer easier. Dentists will have an opportunity to do community service and help patients while also marketing their practice. The community will be aware of mouthguards, and those who participate in athletic activities can make more educated decisions on mouthguard use. PMID- 11851273 TI - Have you considered intraoral imaging? PMID- 11851274 TI - [Luxation of hip prosthesis]. PMID- 11851275 TI - [Anthrax as biological weapon and hospital infection control routines]. PMID- 11851276 TI - [The brave bee of general medicine]. PMID- 11851277 TI - [Screening for DiGeorge syndrome and other genetic diseases]. PMID- 11851278 TI - [Adverse effects of COX-2 inhibitors]. PMID- 11851279 TI - [Methylmalonic acid in diagnosis of vitamin B12 deficiency]. PMID- 11851280 TI - [Compulsory admissions]. PMID- 11851282 TI - [The general practice system and reduction of waiting lists]. PMID- 11851281 TI - [When you become old]. PMID- 11851283 TI - [Admitted to a company]. PMID- 11851285 TI - [Ritual circumcision]. PMID- 11851284 TI - [Forensic psychiatry in past, in present time and in future]. PMID- 11851286 TI - [Personality disorders]. PMID- 11851287 TI - [Cultural perspectives of health and disease]. PMID- 11851288 TI - [Open surgery of abdominal aortic aneurysm]. AB - BACKGROUND: We wanted to evaluate the results after open surgical treatment of abdominal aortic aneurysm. MATERIAL AND METHODS: We present a retrospective survey of 581 patients operated for infrarenal abdominal aortic aneurysm at the Tromso University Hospital from 1980 through 1998. RESULTS: The number of elective operations increased from two in 1980 to 18 in 1998. The frequency of conditions regarded as co-morbidity increased during the observed time period. Mean body mass index for electively operated men increased from 24.2 in 1980-84 to 26.1 in 1995-98 (p < 0.01). Mean aneurysms diameter of those operated electively was 60 mm. The operative (30 days) mortality rate was 7.6% for those operated electively, 9.8% for those with impending rupture, and 40% for those with ruptured aneurysm. INTERPRETATION: We find these results comparable with those presented in other reports. PMID- 11851289 TI - [Incisional hernia after surgery for abdominal aortic aneurysm]. AB - BACKGROUND: Incisional hernia in patients electively operated on the abdominal aorta is common. METHODS: Patients with abdominal aortic aneurysm and with aorto iliac occlusive disease (n = 231) operated at Tromso University Hospital between 1992 and 1998 were identified. Data were obtained from patient files and from a postal questionnaire sent to 172 patients and returned by 140 (85%). RESULTS: Mean follow-up time was 3.5 years; 49 out of 140 patients (35%) had incisional hernia, 42 of these had been operated for aneurysm and seven for occlusive disease. Patients operated for aneurysm had an increased risk of developing incisional hernia (OR 3.1, p = 0.03). Obesity and smoking were associated risk factors. The hernias appeared earlier in patients with aorta aneurysm than in patients with occlusive disease (p < 0.01). A large proportion of the patients reported discomfort due to their hernias. INTERPRETATION: Abdominal aorta aneurysm seems to be associated with alterations in the connective tissue. Incisional hernia is a significant source of morbidity in patients operated for abdominal aortic aneurysm. PMID- 11851290 TI - [Rescue helicopter service in Bodo--advanced emergency service or alternative transportation?]. AB - BACKGROUND: An increase in fatal accidents in helicopter ambulance missions in Norway has put focus on the guidelines for use. METHODS: Ambulance records from the physician-staffed SeaKing rescue helicopter in Bodo, Northern Norway, from 1988 to 1998 were analysed retrospectively. RESULTS: 2,498 ambulance missions carried 2,590 patients. Median time to lift-off was 29 minutes, and one-way median flying time was 26 minutes. Four local communities with a total of 12,000 residents (6.3% of the population served by the helicopter) ordered 70% of the missions. 35% of the patients suffered from cardiovascular disease, 12% were in labour, 3.2% were seriously traumatized, while 20% had minor injuries. A total of 107 patients (4%) received advanced pre-hospital emergency treatment. Thirty eight (1.5%) were intubated and received cardiopulmonary resuscitation, of whom two survived to discharge. Over a period of 11 years, 53 patients were intubated by the anaesthesiologist. Oxygen or intravenous lines as the only treatment effort were given to 72%. INTERPRETATION: It takes about an hour to reach remote locations by rescue helicopter. A substantial amount of the transports could have been carried out by ground ambulance without loss of health benefit. We question the need for an on-board anaesthesiologist. PMID- 11851291 TI - [Tick-borne encephalitis in Norway]. AB - BACKGROUND: The first person reported with tick-borne encephalitis (TBE) in Norway fell ill in 1998; the second patient caught the disease in autumn 1999. Both had been to the island of Tromoy in Aust-Agder county. Searches for TBE antibodies were intensified in persons with encephalitis and a seroprevalence study was carried out. MATERIAL AND METHODS: Sera from persons with encephalitis were tested for IgM and IgG antibodies to TBE virus with enzyme immunoassay and a neutralization test. Stored sera from persons living on Tromoy in 1997-2000 were studied for IgG antibodies. RESULTS: We report three cases of TBE, two had encephalitis in September-October 2000. The third patient, with antibodies to TBE virus found by retrospective testing, had the disease in August 1997. IgG antibodies to TBE virus were found in 3 out of 126 (2.4%) samples from Tromoy. INTERPRETATION: We report the first case of TBE in Norway. Of the first five Norwegian cases, four had been on Tromoy before they fell ill, three of them as tourists. In previous studies, IgG antibodies to TBE virus were found in 0.3-0.4% of persons from different parts of Agder counties. The seroprevalence studies indicate that Tromoy may have a higher incidence of TBE than the rest of the two southern counties. Our results confirm that TBE occurs in the coastal area of southern Norway. PMID- 11851292 TI - [African trypanosomiasis--a rare imported disease]. AB - This year at least nine cases of African trypanosomiasis have occurred among Europeans visiting the Serengeti Park in Tanzania. One of them was a 26-year-old Norwegian woman who was bitten by a tsetse fly in her face. She developed fever, nausea and other symptoms, and had thrombocytopenia and pathological liver values; trypanosoma parasites were demonstrated in her blood. The recommended drug, suramin, was not readily available in Tanzania and she was transported to Oslo. The diagnosis was confirmed. She had no affection of the central nervous system and was treated successfully with suramin only. Increased tourist traffic to exotic destinations may lead to more frequent import of trypanosomiasis to European countries. Drugs to treat the disease must be readily available. PMID- 11851293 TI - [Trypanosomiasis--a real risk for tourists visiting national parks in Tanzania]. AB - African sleeping sickness is no longer a rare disease among tourists visiting national parks in Tanzania. The disease is caused by a parasite, Trypanosoma brucei, which is transmitted by the tsetse fly. Two species infect humans: Trypanosoma brucci gambiense and Trypanosoma brucei rhodesiense; the last form is re-emerging in parts of Africa. Untreated this disease carries a mortality of nearly 100%. This article describes a case of African sleeping sickness in a tourist visiting Tanzania, which was diagnosed at the Nordic Clinic, Dar es Salaam. The most important symptoms, diagnostic investigations as well as the main principles of treatment are described. Patients with this condition need to be admitted and treated at centres with competence in tropical diseases. African sleeping sickness should be kept in mind in tourists returning to their home country with fever after visits to national parks in Eastern Africa. With early treatment, cure is almost certain. The only way to prevent this condition is through protection against bites of the tsetse fly. PMID- 11851294 TI - [Genocide--some medical and psychological aspects]. AB - The Nazi genocide of the Jews was the background for the UN Convention on Genocide in 1949. The number of people murdered according to the definition in the UN Convention in the period 1900-87 has been estimated at 37 million; the real number is probably higher. Genocide is a considerable cause of death, it has severe consequences for survivors, is an irreplaceable loss for surviving relatives and a trauma for the collective identity. Fair trials of the perpetrators is important to the victims and society. The disastrous role of medicine in the Holocaust should be included in educational programmes as a reminder of the vulnerability of the health professions. PMID- 11851295 TI - [Fiberoptic phototherapy for neonatal icterus--comments to a Cochrane report]. AB - Twenty-four studies filled the inclusion criteria for a Cochrane report on the use of fiberoptic phototherapy for neonatal jaundice. Fiberoptic phototherapy lowers serum bilirubin and may have a place in the treatment of neonatal jaundice. In term and near-term infants fiberoptic phototherapy is inferior to conventional phototherapy, whereas in premature infants the effects are comparable. A better effect is achieved when conventional and fiberoptic phototherapy are combined as compared to conventional phototherapy alone. Fiberoptic phototherapy has not been shown to interfere less with parent-infant bonding than conventional phototherapy. There is still a great need for more research on phototherapy for neonatal jaundice. PMID- 11851296 TI - [Etiology and genetics of Alzheimer disease]. AB - BACKGROUND: Alzheimer's disease (AD) constitutes more than 50% of all dementias. The diagnosis is mainly based on clinical criteria and a definitive diagnosis of AD is made post-mortem with identification of amyloid plaques and neurofibrillary tangles. A small proportion of the patients are under the age of 60 at diagnosis, known as early-onset AD, and most of these cases have an evident genetic component. Aging is the most important risk factor for developing late-onset AD, but also genetic polymorphisms and many environmental conditions play a part in the development of this multifactorial disease. METHODS: The Medline database was searched for "Alzheimer's and genetics". Histologic data were kindly provided from our hospital's department of pathology. RESULTS AND INTERPRETATION: We consider most of the proved etiological factors, especially the three genetic loci which have been shown to be associated with early-onset AD: amyloid precursor protein (APP) gene, presenilin (PS)-1 and PS-2 genes. Mutations in the PS-1 gene at chromosome 14 are by far the most frequent genetic cause of AD. However, the large number of mutations makes genetic screening difficult. We also discuss the impact of the different ApoE alleles in developing late-onset AD, in addition to other mutations and polymorphisms. PMID- 11851297 TI - [Models for understanding personality pathology]. AB - BACKGROUND: The prevalence of personality disorders is high and patients with personality disorders are known to be difficult to treat. The assessment of different types of personality disorders has been facilitated by the development of explicit diagnostic criteria. However, the diagnostic manuals have tended to be atheoretical, defining and classifying personality disorders mainly on a descriptive level of behaviour and symptoms. There is no consensus on a theoretical understanding of personality pathology. The concept of personality disorder is currently under debate and investigation, theoretically as well as empirically. MATERIAL AND METHODS: This paper presents some of the major theories of personality disorders that are currently discussed in the literature. RESULTS: The models represent different schools of thought, lean on different methodological assumptions, and focus on different aspects of personality. The theories are to varying degrees focused on aetiology, classification and treatment. A major trend is to supplement theories developed within the context of clinical psychiatry with models from normal psychology. INTERPRETATION: Personality disorder is a complex phenomenon. Different theories are probably useful for different aspects of personality pathology and for different groups of personality disorders. More empirical research is needed in order to clarify the validity of the different models. PMID- 11851298 TI - [Experiences with ritual circumcision in Norway]. AB - BACKGROUND: We wanted to provide a survey of the extent and practice of locally performed ritual circumcision of boys born to Muslim parents in Norway. MATERIAL AND METHODS: Newborn boys with Muslim parents in two counties in Norway (Rogaland and Troms) were registered. 56 ritual circumcisions were performed in outpatient departments in the local hospitals during the period under investigation, 11 on newborns, 33 on boys between one and ten, and 12 on boys above ten. RESULTS: The number of circumcised boys in 1999 was 24% of the total number of boys born to Muslim parents that year; in 2000, 48%. Circumcision of boys between one and ten is ten times more time-consuming than circumcision of newborns or boys above ten. INTERPRETATION: An estimated 672 boys are born to Muslim parents in Norway each year. Less than 50% of newborns with Muslim parents are circumcised in local outpatient departments. We recommend that ritual circumcision is restricted to newborns or to boys above ten, and that informed written consent is obtained from boys above ten. PMID- 11851299 TI - [Axel Strom--pioneer of social medicine and administrator]. AB - Dr Axel Strom (1901-85), professor in the University of Oslo from 1940 to 1970, was a leader in Norwegian medicine in the latter half of the 20th century. He qualified in 1926 and in 1936 gained a doctorate with a dissertation on the toxin production of the Corynebacterium diphtheriae. His first appointment as a professor was in hygiene. In 1951 he moved on to public health, a field that he pioneered in Norway and the other Scandinavian countries. As a professor during the German occupation of Norway in the Second World War, he joined the university's resistance against the Nazi authorities' attempts at taking control. When the war was over he became deeply involved in research on the impact of war on health. At a time when the study of the impact of lifestyle factors was still in its infancy, he suggested that the war-induced reduction in dietary fat consumption might be the cause of observed lower cardiovascular mortality. Of more practical importance were the studies he initiated of the mainly psychological late-onset effects of traumas suffered by prisoners in German camps, seamen, soldiers and other exposed groups. In this area, too, he was an early explorer, of what has come to be known as post-traumatic stress disorder. His efforts led to improved war pension entitlements for the victims. Over the years, exposed groups became his major professional interest as a public health specialist. In his academic work, dr Strom also pioneered medical ethics, care for the elderly, legislation on abortion, and the rapidly expanding field of the medical basis for social security benefits. As a practising physician he was in the vanguard of occupational medicine and other kinds of preventive medicine. What brought him most recognition was, however, his leading role over many years in the Norwegian Medical Association and in the University of Oslo. He served as chairman of the Junior Hospital Doctors Association, president of the Norwegian Medical Association and chairman of the Federation of Norwegian Professional Associations. He was elected dean of the Faculty of Medicine and vice-rector of the University of Oslo, in addition to a host of other expert assignments and official roles: He was renowned for his hard work and exerted great influence in many quarters. PMID- 11851300 TI - [Disease and responsibility for one's own actions--history of forensic expertise]. AB - In the 19th century, a general rule of exemption from jural imputability for the mentally deranged was codified in Norwegian criminal law. The legislative measure was not phrased in medical terminology; nevertheless, the ground was cleared for increasing usage of medical expertise in criminal proceedings. The medical community in Norway held diverse opinions as to the role of forensic medicine/psychiatry. This article explores the debate within the profession from the vantage point a homicide case from 1874 in which expert opinions were sought from a country doctor, the director of a state asylum, and from the professors of the national university's faculty of medicine. The various opinions testify to the elasticity of the jural notion of imputability and how inaccurate it would be to label positions in this highly contested field as either progressive or reactionary: 19th century notions of mental illness were as complex as they were context-sensitive. PMID- 11851301 TI - [Technology and culture--HbA1c, self care and type 1 diabetes]. AB - It has been argued that medical technologies do more than simply generate representations of reality. Like other technologies they act on the situations in which they are used. This article is based on ethnographic fieldwork carried out among young adults with type 1 diabetes in Oslo, Norway. An important test in the treatment of type 1 diabetes, the glucosylated haemoglobin test (HbA1c), is discussed as a technology, as a theme in the clinical dialogue, and as a component of the self-management regime. The test gives an estimate of average blood sugar levels and thus serves to represent the quality of the patient's everyday self-management. In the light of the ethics of responsibility established by self-management as a treatment regime, we suggest that the HbA1c test can assume the function of a sign indicating moral qualities in the patient, and that this has an impact on the clinical dialogue. This test-as-sign offers an illustration of how bioscience technologies may enter into people's understanding of themselves in what has been termed a "risk society". PMID- 11851302 TI - [Information campaigns in the fight against tuberculosis until about 1940]. AB - BACKGROUND: Already before the advent of vaccination and antibiotics in Norway in the 1940s, mortality from tuberculosis was heavily reduced. Initiatives from central and local authorities, the efforts of housewives in the homes and the activity of voluntary organizations were attempts to obtain this result. This article focuses on information campaigns staged by the National Association Against Tuberculosis between 1910 and 1920. MATERIAL: Annual reports from the National Association, the Association's journal, the archive from the Lyster Sanatorium in the Bergen Town Archives, and earlier literature on this theme. RESULTS: The Association Against Tuberculosis organised extensive information campaigns through lectures, leaflets, its journal, and "travelling teachers", targeting the general public as well as defined groups. Men were especially admonished to stop spitting, women to keep the home clean, and patients to follow doctor's instructions. INTERPRETATION: The information campaigns were part of comprehensive efforts made by voluntary organizations, public authorities, doctors and housewives to reduce the mortality of tuberculosis before the advent of vaccination. Higher standards of living also contributed to goal attainment. PMID- 11851303 TI - [Voluntarily childless women--wherein lies the problem?]. AB - BACKGROUND: Most present-day Norwegian women choose to have children. This article explores the experience of the small group of voluntarily childless, or child-free, women. MATERIAL AND METHODS: This study is substantially based upon a small number of indepth interviews conducted in 1999 and 2000. The findings are not statistically significant; the objective is to highlight some themes and demonstrate different ways of experiencing and explaining a life without children. All respondents either lived or had lived in heterosexual relationships; at the time of the interviews their marital status were single, divorced, remarried, or living with a partner without being married. All were well-educated city-dwellers. RESULTS: According to their own accounts, the women lived happily without children, though problems arose in their meeting with others (family, friends, colleagues) who did not accept their choice of life style. INTERPRETATION: Modern reproductive medicine has improved the predicament of women who want children but need assistance in order to give birth. On the other hand, women who do not want children find that their choice is criticised and that people around them fail to take it seriously. PMID- 11851304 TI - [The symbolic power of bodily secretions]. AB - Are ethical and political questions discussed more in aesthetic than in social terms? Political debates in contemporary art are often expressed through the use of symbolic power structures that are related to bodily secretions and products. By visualizing the less delicate parts of the organic body, contemporary artists focus on the vulnerability of the human body and its illness and pain. If human suffering is aestheticized: is this a question of form or a question of creating a cultural cover-up? Bodily secretions are associated with a range of complex social and cultural meanings and symbols. Metaphors related to body products may function as markers of ethnic, religious, social and sexual differences. In postmodern cultural criticism, the symbolic power of body metaphors is of great importance. PMID- 11851305 TI - [Young gay men and absence of adequate signs and cultural images]. AB - This article is based on a qualitative, exploratory HIV-related study among young gay men in Norway. In addition to generating HIV-related knowledge, the study provided insights into phenomena associated with the general cultural conditions under which young gay men grow up. Those who are not yet members of gay communities or friendship groups have to rely on the public discourse for their acquisition of knowledge and signs relating to sex and sexuality. However, among the signs and images available through this discourse, only a few mediate a homosexual reality. Furthermore, of the few that exist, hardly any can be found that is not derived from the heterosexual domain. In effect this implies that young gay men will have to make sense of their lives and their sexuality through signs and images that basically mediate a reality different from theirs. In encounters between health workers and young gay men, knowledge about and awareness of such processes may be vital. PMID- 11851306 TI - [Ritual circumcision in Norway]. PMID- 11851307 TI - [False positive serological test in suspected borreliosis]. PMID- 11851308 TI - [Poisonous mushrooms--lot of myths, few facts]. PMID- 11851312 TI - Peptide analysis by rapid, orthogonal technologies with high separation selectivities and sensitivities. AB - This article examines the current status of peptide analysis by orthogonal micro /nano-separation strategies, with emphasis on the complementary use of high performance capillary liquid chromatography (micro-HPLC), capillary zonal electrophoresis (HPCZE), open tubular capillary electrochromatography (ot -CEC) and packed capillary electrochromatography (p -CEC). The ability to interface these techniques with mass spectroscopic (MS) procedures has enabled substantial progress to be made in the analysis of very small quantities of peptides, as well as proteins and other bio-macromolecules. As a consequence, the staged application of these high resolution techniques as part of the standardisation of biological products via robust, sensitive protocols is rapidly becoming a reality. Recent conceptual and theoretical advances have also allowed improved levels of prediction and optimisation of these procedures. Since significant differences in selectivity can be achieved with micro-HPLC, HPCZE and HPCEC respectively, collectively these sophisticated techniques provide unprecedented opportunities for the rapid, orthogonal and sensitive separation of complex mixtures of peptides and proteins. Several advantages of using these technologies in tandem are highlighted. PMID- 11851313 TI - From combinatorial libraries to MHC ligand motifs, T-cell superagonists and antagonists. AB - Complete experimental data sets of HLA-ligand motifs and T-cell recognition patterns can be derived from combinatorial peptide libraries. These data provide the exact molecular basis for a fast development of synthetic vaccines, T-cell superagonists and non-peptide antagonists. Patient-specific peptides, peptidomimetics and vaccines of highest reactivity can be derived directly from the data sets via our prediction programme EPIPREDICT. The resulting lead structures may be developed into valuable diagnostics and therapeutic tools for the treatment of viral infections, autoimmune diseases and tumors. As one example, antibody and T cell recognition in the intestinal auto-immune disease, coeliac disease was investigated in more detail concerning the deamidation of gamma-gliadin peptides by tissue transglutaminase 9tTG) leading to autoreactive peptides specific for HLA-DQA1*0501, DQB1*0201. PMID- 11851314 TI - Chemoselective ligation and antigen vectorization. AB - The interest in cocktail-lipopeptide vaccines has now been confirmed by phase I clinical trials: highly diversified B-, T-helper or cytotoxic T-cell epitopes can be combined with a lipophilic vector for the induction of B- and T-cell responses of predetermined specificity. With the goal of producing an improved vaccine that should ideally induce a multispecific response in non-selected populations, increasing the diversity of the immunizing mixture represents one of the most obvious strategies.The selective delivery of antigens to professional antigen presenting cells represents another promising approach for the improvement of vaccine efficacy. In this context, the mannose-receptor represents an attractive entry point for the targeting to dendritic cells of antigens linked to clustered glycosides or glycomimetics. In all cases, highly complex but fully characterized molecules must be produced. To develop a modular and flexible strategy which could be generally applicable to a large set of peptide antigens, we elected to explore the potentialities of chemoselective ligation methods. The hydrazone bond was found particularly reliable and fully compatible with sulphide ligation. Hydrazone/thioether orthogonal ligation systems could be developed to account for the nature of the antigens and the solubility of the vector systems. PMID- 11851315 TI - Tandem peptide ligation for synthetic and natural biologicals. AB - We describe the concept and methods of peptide ligation and tandem peptide ligation for preparing synthetic and natural biologicals. Peptide ligation is a segment coupling method for free peptides or proteins through an amide bond without the use of a coupling reagent or a protecting group scheme. Because unprotected peptides or proteins prepared from either a chemical or biochemical source are being used as building blocks, the ligation removes the size limitation for peptide and protein synthesis. A key feature of the peptide ligation is that the coupling reaction is orthogonal, i.e. it is specific to a particular alpha-amino terminus (NT). This NT-amino acid-specific feature permits the development of a tandem peptide ligation method employing three unprotected peptide segments containing different NT-amino acids to form consecutively two amide bonds, an Xaa-SPro (thiaproline) and then an Xaa-Cys. This strategy was tested in peptides ranging from 28 to 70 amino acid residues, including analogues of somatostatins and two CC-chemokines MIP-1alpha and MIP-1beta. The thiaproline replacements in these peptides and proteins did not result in altered biological activity. By eliminating the protecting group scheme and coupling reagents, tandem ligation of multiple free peptide segments in aqueous solutions enhances the scope of protein synthesis and may provide a useful approach for preparing protein biologicals and synthetic vaccines. PMID- 11851316 TI - Manipulation of epitope function by modification of peptide structure: a minireview. AB - We have explored various approaches to modify the immunrecognition of linear peptides representing sequential or continuous topographic B-cell or T-cell epitopes. For these studies, epitopes from herpes simplex virus (HSV) glycoprotein D (gD) and from mucin 1 and mucin 2 glycoproteins or T-cell epitopes from 16 kDa and 38 kDa proteins of Mycobacterium tuberculosis were selected. To increase antigenicity and immunogenicity we have prepared cyclic and chimaeric peptide variants as well as epitope peptides with altered flanking regions and epitope-carrier conjugates containing multiple epitope copies. PMID- 11851317 TI - Antigenicity and immunogenicity of synthetic peptides. AB - The ability of a peptide to react specifically with the functional binding site of a complementary antibody is known as its antigenic reactivity or antigenicity. Our understanding of peptide antigenicity has improved considerably in recent years mainly through the X-ray crystallographic analysis of peptide-monoclonal antibody complexes. This knowledge is obtained along reductionist lines by turning the biological question of antigen recognition into the purely chemical phenomenon of protein-peptide interactions described in terms of atomic forces and non-covalent bonds. This makes it possible to improve the degree of steric complementarity between a peptide and a single monoclonal antibody and thus to improve the peptide's antigenicity following structure-based rational design principles. The situation is quite different with immunogenicity which is the ability of the peptide to induce an immune response in a competent host. Whereas antigenicity can be reduced to the level of chemistry, such a reduction is not achievable in the case of immunogenicity which depends on many complex interactions with various elements of the host immune system. These cellular and regulatory mechanisms cannot be controlled by adjusting the structure of the peptide in a predetermined manner. For this reason, it is not possible to develop a synthetic peptide vaccine using molecular design principles. PMID- 11851318 TI - The development of a mimotope-based synthetic peptide vaccine against respiratory syncytial virus. AB - Respiratory syncytial virus (RSV) is the most important cause of bronchiolitis and pneumonia in infants and young children worldwide and the development of a synthetic peptide epitope-based vaccine to induce virus-neutralising antibodies against RSV would seem to be a valid approach to the production of an effective vaccine against infection.A combinatorial solid-phase peptide library has been screened with a virus-neutralising, protective monoclonal antibody (MAb19) directed towards a conserved and conformationally-determined epitope of the Fusion (F) protein of the virus. Two of the sequences identified from the peptide library using MAb19 reacted specifically with the antibody and amino acid substitution experiments identified four sequences from one of the mimotopes which showed increased reactivity with MAb19. Immunisation of BALB/c mice with these mimotopes, presented as MAPs, resulted in the induction of anti-peptide antibodies that inhibited the binding of MAb19 to the virus and neutralised viral infection in vitro, with titres equivalent to those in sera from RSV-infected animals. Following RSV challenge of mimotope-immunised mice, a significant reduction in the titre of virus and a greatly reduced cell infiltration into the lungs of immunised mice compared to that in controls was observed. The induction of virus-specific cytotoxic T-lymphocyte responses as well as virus-specific antibodies are likely to be necessary in an effective vaccine. The incorporation of a peptide representing a CTL epitope from the M2 protein of the virus together with peptides inducing T-helper and anti-mimotope responses in a peptide cocktail vaccine resulted in a more effective clearance of the virus from immunised, challenged mice than peptide-induced humoral or cellular immunity alone. PMID- 11851319 TI - Synthetic peptide-based vaccine and diagnostic system for effective control of FMD. AB - We have designed synthetic peptides corresponding to two different regions of the genome of foot-and-mouth disease virus (FMDV) that are effective as (a) a vaccine or (b) a diagnostic reagent which differentiates convalescent from vaccinated animals, respectively. The peptide vaccine is based on a sequence from the prominent G-H loop of VP1, one of the four capsid proteins. The sequence was optimized by the inclusion of a cyclic constraint and adjoining sequences, and broader immunogenicity was obtained by the incorporation of consensus residues at hypervariable positions. The peptide also included a promiscuous T-helper epitope for effective immunogenicity in outbred populations of large animals.The diagnostic reagent, a peptide based on non-structural (NS) protein 3B, is used in immuno-assays for the detection of antibodies. Antibodies to this NS protein are present in the sera of infected animals but not in the sera of vaccinated animals. The VP1 peptide can be used in complementary immuno-assays for confirmation of NS test results and to monitor for vaccination. This system for differential diagnosis is important to establish the disease-free status of a country. PMID- 11851320 TI - Mapping of natural anti-factor VIII antibodies in plasma pools from healthy donors: use of rationally designed synthetic peptides. AB - Inhibitor antibodies of blood coagulation factor VIII (FVIII) impair FVIII replacement therapy, constituting a serious complication in haemophilic patients. anti-FVIII antibodies may also develop in a variety of disease-associated autoimmunity. Mapping of human FVIII inhibitors in haemophilia A or autoantibody origin have delineated three major clusters of B-cell inhibitory epitopes (domain A2, A3 and C2). Inhibitory and non-inhibitory FVIII antibodies have also been described in plasma of healthy donors and pools of immunoglobulins. The purpose of this study was to use synthetic FVIII-peptides to more closely define regions of the molecule targeted by natural anti-FVIII antibodies. Predictive algorithms were used for defining the positions of potential continuous epitopes. To investigate the presence of peptide-reactive antibodies in normal plasma pools of healthy donors, a plasma fraction (Cohn fraction II+III) containing all IgG subclasses was purified by affinity chromatography on peptide-Sepharose columns. The results of ELISAs and Western blotting experiments (with the selected peptides and well-defined recombinant FVIII thrombin fragments) confirmed the reaction specificities of the affinity-purified human antibodies. For each IgG preparation, the isotopic subclass was also determined. In the clotting assay, several IgG preparations showed neutralising activity in a dose-dependent manner. Our observations support the recent hypothesis that FVIII inhibitors in haemophilia A and autoimmune disease may originate from the proliferation of natural FVIII-specific B-cell clones. PMID- 11851321 TI - Synthetic peptide vaccines: unexpected fulfillment of discarded hope? AB - In the early eighties it was realized that the ultimate vaccine would be a synthetic peptide. Major efforts were put into the development of a synthetic vaccine for foot-and-mouth disease virus (FMDV) for which even today no alternative exists besides the classical vaccine based on inactivated virus. Despite impressive progress, a peptide vaccine that could match the classical vaccine with respect to efficacy (i.e. full protection of all animals after a single vaccination) has not materialized. This has led to the belief that synthetic vaccines were not possible. However, in the early nineties we developed a synthetic peptide vaccine for canine parvovirus that did match the classical vaccine based on inactivated virus (i.e. protected all animals). Based on the difference of FMDV (an RNA virus) and canine parvovirus (a DNA virus), we suggested that in the case of FMDV, more than one antigenic site should be used, instead of the single one used previously. In our opinion multiple sites are necessary to prevent the development of escape mutants of FMDV. Unfortunately, the additional sites of FMDV are highly discontinuous. Until recently it was impossible to reconstruct these sites in the form of synthetic peptides. In the past few years, new methods have been developed that allow recombination of such sites into synthetic molecules. If successfully applied to FMDV, synthetic peptide vaccines and many others may become feasible in the near future. Moreover, the ability to mimic complex discontinuous sites by synthetic peptides will have a major impact on the rapidly developing area of therapeutic vaccines. PMID- 11851322 TI - Peptide-based synthetic recombinant vaccines with anti-viral efficacy. AB - Synthetic recombinant vaccines are constructs in which a synthetic oligonucleotide coding for a protective epitope is inserted into an adequate gene for expression of the epitope. We report the results obtained using recombinant flagella of Salmonella vaccine strain expressing epitopes of influenza virus or of the parasite Schistosoma mansoni. In the case of influenza virus, three conserved epitopes of the haemagglutinin and the nucleoprotein of the virus inducing B- and T-cell immune response, were expressed and the flagella were used for intranasal immunization without any adjuvant. Both humoral and cellular immune responses specific to the virus induced in mice cross-strain long-term protection against challenge infection. Aged mice were also able to resist infection. For the design of a human influenza vaccine, epitopes recognized by the HLAs prevalent in Caucasian populations were used, and the resulting vaccine was evaluated in human/mouse radiation chimaera in which human PBMC are functionally engrafted. The vaccinated mice demonstrated efficient clearance of the virus after challenge and resistance to lethal infection. In the case of the parasitic disease schistosomiasis, a 14-residue peptide denoted 9B peptide 1 was expressed in the flagella. Intranasal vaccination of mice with this construct, without the use of adjuvant, resulted in 40% protection against challenge infection. PMID- 11851323 TI - Towards Alzheimer's beta-amyloid vaccination. AB - Beta-amyloid pathology, the main hallmark of Alzheimer's disease (AD), has been linked to its conformational status and aggregation. We recently showed that site directed monoclonal antibodies (mAbs) towards the N-terminal region of the human beta-amyloid peptide bind to preformed beta-amyloid fibrils (Abeta), leading to disaggregation and inhibition of their neurotoxic effect. Here we report the development of a novel immunization procedure to raise effective anti-aggregating amyloid beta-protein (AbetaP) antibodies, using as antigen filamentous phages displaying the only EFRH peptide found to be the epitope of these antibodies. Due to the high antigenicity of the phage no adjuvant is required to obtain high affinity anti-aggregating IgG antibodies in animals model, that exhibit identity to human AbetaP. Such antibodies are able to sequester peripheral AbetaP, thus avoiding passage through the blood brain barrier (BBB) and, as recently shown in a transgenic mouse model, to cross the BBB and dissolve already formed beta amyloid plaques. To our knowledge, this is the first attempt to use as a vaccine a self-anti-aggregating epitope displayed on a phage, and this may pave the way to treat abnormal accumulation-peptide diseases, such as Alzheimer's disease or other amyloidogenic diseases. PMID- 11851324 TI - Immunological characterization of peptide mimetics of carbohydrate antigens in vaccine design strategies. AB - Targeting antigens which cannot be readily addressed by genetic vectors is a major challenge in vaccine design. The inter-conversion of carbohydrate antigens into peptide mimetic forms provides a means to broaden the immune response to carbohydrate antigens. Peptides that mimic carbohydrate antigens offer new possibilities to augment immune responses to such antigens that include inducing carbohydrate reactive T-cell responses. Peptide mimeotopes can be formulated in a variety of ways that include multiple antigen peptides (MAP) and as DNA vaccines that prime for different antibody isotypes. On the immunological side we observe that: (i) depending on the immunogen formulation peptide mimetics can be processed by either CD5+ or CD5-B cells; (ii) peptide mimeotope immunization can induce cross-reactive responses to multiple carbohydrate forms; (iii) priming with peptide mimeotopes can enhance carbohydrate immune responses upon boosting and (iv) immunization with peptide mimeotopes can induce carbohydrate reactive T cells. PMID- 11851325 TI - Long synthetic peptides as biologically active proteins: the example of the chemokines. AB - Chemokines constitute an expanding protein family of over 40 members which exhibit a wide variety of biological activities and are involved in many normal physiological processes, such as cellular migration, differentiation and activation, but also in pathological situations, such as inflammation and metastasis. Over the last few years, we have developed methods to manufacture long synthetic peptides of up to 130 residues, and to achieve the formation of native-like cysteine pairings. This ability prompted us to undertake the total chemical synthesis of chemokines. So far, we have successfully produced over 30 chemokine species, which exhibit biological activities similar to, or greater than, those reported by others. Chemical synthesis offers a clear advantage over recombinant technologies for the introduction of fluorochromes and haptens at molecularly defined positions. In addition, approval of chemically synthesized products for use in humans is straightforward compared with material produced by biological methods. PMID- 11851326 TI - Synthetic peptides as functional mimics of a viral discontinuous antigenic site. AB - Functional reproduction of discontinuous antigenic site D of foot-and-mouth disease virus (FMDV) has been achieved by means of synthetic peptide constructions that integrate into a single molecule each of the three protein loops that define the antigenic site. The site D mimics are designed on the basis of the X-ray structure of FMDV type C-S8c1 with the aid of molecular dynamics, so that the five residues assumed to be involved in antigenic recognition are located on the same face of the molecule, exposed to solvent and defining a set of native-like distances and angles. The designed site D mimics are disulphide linked heterodimers that consist of a larger unit containing VP2(71-84), followed by a polyproline module and by VP3(52-62), and a smaller unit corresponding to VP1(188-194). Guinea pig antisera to the peptides recognize the viral particle and compete with site D-specific monoclonal antibodies, while inoculation with a simple (non-covalently bound) admixture of the three VP1-VP3 sequences yields no detectable virus-specific serum conversion. Similar results have been reproduced in two cattle. Antisera to the peptides are also moderately neutralizing of FMDV in cell culture and partially protective of guinea pigs against challenge with the virus. These results demonstrate functional mimicry of the discontinuous site D by the peptides, which are therefore obvious candidates for a multicomponent peptide-based vaccine against FMDV. PMID- 11851327 TI - The development of multi-epitope vaccines: epitope identification, vaccine design and clinical evaluation. AB - We have developed efficient methods for epitope identification and vaccine design. Our process for epitope selection based on the combined use of motif analyses, binding assays and immunogenicity evaluations is described. We also describe how the projected population coverage and vaccine design can be optimized. Finally, it is discussed how vaccine potency is evaluated by immunogenicity and antigenicity assays. PMID- 11851329 TI - Designing peptide vaccines for cellular cross-presentation. AB - Synthetic peptides are safe and relatively cheap vaccine components. However, the efficiency of peptide vaccines is limited by peptide interaction with non professional antigen-presenting cells, which may hamper induction of productive T cell responses. This paper argues that peptide vaccines should be modified for exclusive uptake by cells with the capacity to prime T-cell responses. Moreover, design of peptide vaccines should take intracellular antigen processing into account and exploit cellular mechanisms of proteolysis, transport and HLA class I assembly of antigenic peptides to enhance efficiency of T-cell priming and stimulation. PMID- 11851328 TI - Circumventing T-cell tolerance to tumour antigens. AB - During past decades, many attempts have been made to induce or enhance tumour specific T-cell immunity in cancer patients by vaccination. However, it has become apparent that in a large number of cases the naturally occurring tumour specific T-cell repertoire is of low affinity and therefore inefficient in mediating tumour rejection. Because of the potential therapeutic value of high affinity TCRs with tumour/lineage specificities, we set out to develop a number of new technologies that can be used to create improved tumour-specific T-cell immunity. These strategies entail: (i) the efficient expansion of low affinity T cells specific for self antigens through the use of variant peptides with improved TCR-binding characteristics; (ii) a retroviral library-based technology to improve the affinity of (self-specific) T-cell receptors in vitro, and (iii) proof of principle for the feasibility of TCR gene transfer as a means to generate T-cell populations with a desired antigen-specificity in vivo. Collectively this toolbox should allow us to create improved T-cell receptors for human tumour antigens, which can subsequently be used to impose tumour-reactivity on to peripheral T cells. PMID- 11851330 TI - Structure-based design and evaluation of MHC class II binding peptides. AB - Structural information regarding binding of peptides to the major histocompatibility complex (MHC) class II molecule is of great use for the design of compounds that intervene in the interaction between the MHC-peptide-T-cell receptor (TCR) complex. These compounds can be applied in the treatment of T-cell mediated auto-immune disease for specific modulation of the disease process. In case no crystal structure of the MHC molecule is available, homology models of the MHC molecule can be of importance. Here we describe the construction of a homology model of the MHC class II molecule and binding of the peptide, that are involved in experimental auto-immune encephalomyelitis, a rat model for human multiple sclerosis. The validity of the model was investigated using experimental data of peptides binding to this MHC molecule. PMID- 11851331 TI - Stability and CTL-activity of P40/ELA melanoma vaccine candidate. AB - The decapeptide ELA (ELAGIGILTV), a Melan-A/MART-1 antigen immunodominant peptide analogue, is an interesting melanoma vaccine candidate alone or in combination with other tumour antigens. P40, the recombinant outer membrane protein A of Klebsiella pneumoniae (kpOmpA), was recently shown to target dendritic cells and to induce peptide-specific CTLs. Here we investigated the adjuvant role of P40 mixed or chemically conjugated to ELA. This compound is an N-terminal glutamic acid-containing peptide. However, it has been reported that the amino group and the gamma-carboxylic group of glutamic acids easily condense to form pyroglutamic derivatives. Usually, to overcome this stability problem, peptides of pharmaceutical interest were developed with a pyroglutamic acid instead of N terminal glutamic acid, without loss of pharmacological properties. Unfortunately, the pyroglutamic acid derivative (PyrELA) as well as the N terminal acetyl capped derivative (AcELA) failed to elicit CTL activity when mixed with P40 adjuvant protein. Despite the apparent minor modifications introduced by PyrELA and AcELA, these two derivatives have probably lower affinity than ELA for the class I Major Histocompatibility Complex. Furthermore, this stability problem is worse in the case of clinical grade ELA, produced as an acetate salt, like most of the pharmaceutical grade peptides. We report here that the hydrochloride shows a higher stability than the acetate and may be suitable for use in man. PMID- 11851332 TI - Molecular modelling of the C-terminal domains of factor H of human complement: a correlation between haemolytic uraemic syndrome and a predicted heparin binding site. AB - Factor H (FH) of the complement system acts as a regulatory cofactor for the factor I-mediated cleavage of C3b and binds to polyanionic substrates. FH is composed of 20 short consensus/complement repeat (SCR) domains. A set of 12 missense mutations in the C-terminal domains between SCR-16 to SCR-20 is associated with haemolytic uraemic syndrome. Recent structural models for intact FH permit the molecular interpretation of these amino acid substitutions. As all nine SCR-20 substitutions correspond to normal amounts of FH in plasma, and were localised in mostly surface-exposed positions, these are inferred to lead to a functional defect in FH. The nine substitutions occur in the same spatial region of SCR-20. As this surface coincides with conserved basic residues in the C terminal SCR-20 domain, the substitutions provide direct evidence for a polyanionic binding surface. The positions of these conserved basic residues coincide with those of heparin-binding residues in the crystal structure of the acidic fibroblast growth factor-heparin complex. A tenth substitution and another conserved basic residue in SCR-19 are proximate to this binding site. As the remaining FH substitutions could also be correlated with their proximity to conserved basic residues, haemolytic uraemic syndrome may result from a failure of FH to interact with polyanions at cell surfaces in the kidney. PMID- 11851333 TI - Residues participating in the protein folding nucleus do not exhibit preferential evolutionary conservation. AB - To what extent does natural selection act to optimize the details of protein folding kinetics? In an effort to address this question, the relationship between an amino acid's evolutionary conservation and its role in protein folding kinetics has been investigated intensively. Despite this effort, no consensus has been reached regarding the degree to which residues involved in native-like transition state structure (the folding nucleus) are conserved. Here we report the results of an exhaustive, systematic study of sequence conservation among residues known to participate in the experimentally (Phi-value) defined folding nuclei of all of the appropriately characterized proteins reported to date. We observe no significant evidence that these residues exhibit any anomalous sequence conservation. We do observe, however, a significant bias in the existing kinetic data: the mean sequence conservation of the residues that have been the subject of kinetic characterization is greater than the mean sequence conservation of all residues in 13 of 14 proteins studied. This systematic experimental bias gives rise to the previous observation that the median conservation of residues reported to participate in the folding nucleus is greater than the median conservation of all of the residues in a protein. When this bias is corrected (by comparing, for example, the conservation of residues known to participate in the folding nucleus with that of other, kinetically characterized residues) the previously reported preferential conservation is effectively eliminated. In contrast to well-established theoretical expectations, both poorly and highly conserved residues are apparently equally likely to participate in the protein-folding nucleus. PMID- 11851334 TI - Dimer formation and transcription activation in the sporulation response regulator Spo0A. AB - The response regulator Spo0A is the master control element in the initiation of sporulation in Bacillus subtilis. Like many other multi-domain response regulators, the latent activity of the effector, C-terminal domain is stimulated by phosphorylation on a conserved aspartic acid residue in the regulatory, N terminal domain. If a threshold concentration of phosphorylated Spo0A is achieved, the transcription of genes required for sporulation is activated, whereas the genes encoding stationary phase sentinels are repressed, and sporulation proceeds. Despite detailed genetic, biochemical and structural characterisation, it is not understood how the phosphorylation signal in the receiver domain is transduced into DNA binding and transcription activation in the distal effector domain. An obstacle to our understanding of Spo0A function is the uncertainty concerning changes in quaternary structure that accompany phosphorylation. Here we have revisited this question and shown unequivocally that Spo0A forms dimers upon phosphorylation and that the subunit interactions in the dimer are mediated principally by the receiver domain. Purified dimers of two mutants of Spo0A, in which the phosphorylatable aspartic acid residue has been substituted, activate transcription from the spoIIG promoter in vitro, whereas monomers do not. This suggests that dimers represent the activated form of Spo0A. PMID- 11851335 TI - Engineered improvements in DNA-binding function of the MATa1 homeodomain reveal structural changes involved in combinatorial control. AB - We have engineered enhanced DNA-binding function into the a1 homeodomain by making changes in a loop distant from the DNA-binding surface. Comparison of the free and bound a1 structures suggested a mechanism linking van der Waals stacking changes in this loop to the ordering of a final turn in the DNA-binding helix of a1. Inspection of the protein sequence revealed striking differences in amino acid identity at positions 24 and 25 compared to related homeodomain proteins. These positions lie in the loop connecting helix-1 and helix-2, which is involved in heterodimerization with the alpha 2 protein. A series of single and double amino acid substitutions (a1-Q24R, a1-S25Y, a1-S25F and a1-Q24R/S25Y) were engineered, expressed and purified for biochemical and biophysical study. Calorimetric measurements and HSQC NMR spectra confirm that the engineered variants are folded and are equally or more stable than the wild-type a1 homeodomain. NMR analysis of a1-Q24R/S25Y demonstrates that the DNA recognition helix (helix-3) is extended by at least one turn as a result of the changes in the loop connecting helix-1 and helix-2. As shown by EMSA, the engineered variants bind DNA with enhanced affinity (16-fold) in the absence of the alpha 2 cofactor and the variant alpha 2/a1 heterodimers bind cognate DNA with specificity and affinity reflective of the enhanced a1 binding affinity. Importantly, in vivo assays demonstrate that the a1-Q24R/S25Y protein binds with fivefold greater affinity than wild-type a1 and is able to partially suppress defects in repression by alpha 2 mutants. As a result of these studies, we show how subtle differences in residues at a surface distant from the functional site code for a conformational switch that allows the a1 homeodomain to become active in DNA binding in association with its cofactor alpha 2. PMID- 11851336 TI - Crystal structure of human cytosolic phosphoenolpyruvate carboxykinase reveals a new GTP-binding site. AB - We report crystal structures of the human enzyme phosphoenolpyruvate carboxykinase (PEPCK) with and without bound substrates. These structures are the first to be determined for a GTP-dependent PEPCK, and provide the first view of a novel GTP-binding site unique to the GTP-dependent PEPCK family. Three phenylalanine residues form the walls of the guanine-binding pocket on the enzyme's surface and, most surprisingly, one of the phenylalanine side-chains contributes to the enzyme's specificity for GTP. PEPCK catalyzes the rate limiting step in the metabolic pathway that produces glucose from lactate and other precursors derived from the citric acid cycle. Because the gluconeogenic pathway contributes to the fasting hyperglycemia of type II diabetes, inhibitors of PEPCK may be useful in the treatment of diabetes. PMID- 11851337 TI - The crystal structure of human MRP14 (S100A9), a Ca(2+)-dependent regulator protein in inflammatory process. AB - Human MRP14 (hMRP14) is a Ca(2+)-binding protein from the S100 family of proteins. This protein is co-expressed with human MRP8 (hMRP8), a homologue protein in myeloid cells, and plays an indispensable role in Ca(2+)-dependent functions during inflammation. This role includes the activation of Mac-1, the beta(2) integrin which is involved in neutrophil adhesion to endothelial cells. The crystal structure of the holo form of hMRP14 was analyzed at 2.1 A resolution. hMRP14 is distinguished from other S100 member proteins by its long C terminal region, and its structure shows that the region is extensively flexible. In this crystal structure of hMRP14, Chaps molecules bind to the hinge region that connects two EF-hand motifs, which suggests that this region is a target binding site of this protein. Based on a structural comparison of hMRP14 with hMRP8 and human S100A12 (hS100A12) that is another homologue protein, the character of MRP8/14 hetero-complex and the functional significance of the flexibility of the C-terminal region of hMRP14 are discussed. PMID- 11851338 TI - Structure of a phage display-derived variant of human growth hormone complexed to two copies of the extracellular domain of its receptor: evidence for strong structural coupling between receptor binding sites. AB - The structure of the ternary complex between the phage display- optimized, high affinity Site 1 variant of human growth hormone (hGH) and two copies of the extracellular domain (ECD) of the hGH receptor (hGHR) has been determined at 2.6 A resolution. There are widespread and significant structural differences compared to the wild-type ternary hGH hGHR complex. The hGH variant (hGH(v)) contains 15 Site 1 mutations and binds>10(2) tighter to the hGHR ECD (hGH(R1)) at Site 1. It is biologically active and specific to hGHR. The hGH(v) Site 1 interface is somewhat smaller and 20% more hydrophobic compared to the wild-type (wt) counterpart. Of the ten hormone-receptor H-bonds in the site, only one is the same as in the wt complex. Additionally, several regions of hGH(v) structure move up to 9A in forming the interface. The contacts between the C-terminal domains of two receptor ECDs (hGH(R1)- hGH(R2)) are conserved; however, the large changes in Site 1 appear to cause global changes in the domains of hGH(R1) that affect the hGH(v)-hGH(R2) interface indirectly. This coupling is manifested by large changes in the conformation of groups participating in the Site 2 interaction and results in a structure for the site that is reorganized extensively. The hGH(v)- hGH(R2) interface contains seven H-bonds, only one of which is found in the wt complex. Several groups on hGH(v) and hGH(R2) undergo conformational changes of up to 8 A. Asp116 of hGH(v) plays a central role in the reorganization of Site 2 by forming two new H-bonds to the side-chains of Trp104(R2) and Trp169(R2), which are the key binding determinants of the receptor. The fact that a different binding solution is possible for Site 2, where there were no mutations or binding selection pressures, indicates that the structural elements found in these molecules possess an inherent functional plasticity that enables them to bind to a wide variety of binding surfaces. PMID- 11851339 TI - Dissection of the energetic coupling across the Src SH2 domain-tyrosyl phosphopeptide interface. AB - Src Homology (SH2) domains play critical roles in signaling pathways by binding to phosphotyrosine (pTyr)-containing sequences, thereby recruiting SH2 domain containing proteins to tyrosine-phosphorylated sites on receptor molecules. Investigations of the peptide binding specificity of the SH2 domain of the Src kinase (Src SH2 domain) have defined the EEI motif C-terminal to the phosphotyrosine as the preferential binding sequence. A subsequent study that probed the importance of eight specificity-determining residues of the Src SH2 domain found two residues which when mutated to Ala had significant effects on binding: Tyr beta D5 and Lys beta D3. The mutation of Lys beta D3 to Ala was particularly intriguing, since a Glu to Ala mutation at the first (+1) position of the EEI motif (the residue interacting with Lys beta D3) did not significantly affect binding. Hence, the interaction between Lys beta D3 and +1 Glu is energetically coupled. This study is focused on the dissection of the energetic coupling observed across the SH2 domain-phosphopeptide interface at and around the +1 position of the peptide. It was found that three residues of the SH2 domain, Lys beta D3, Asp beta C8 and AspCD2 (altogether forming the so-called +1 binding region) contribute to the selection of Glu at the +1 position of the ligand. A double (Asp beta C8Ala, AspCD2Ala) mutant does not exhibit energetic coupling between Lys beta D3 and +1 Glu, and binds to the pYEEI sequence 0.3 kcal/mol tighter than the wild-type Src SH2 domain. These results suggest that Lys beta D3 in the double mutant is now free to interact with the +1 Glu and that the role of Lys beta D3 in the wild-type is to neutralize the acidic patch formed by Asp beta C8 and AspCD2 rather than specifically select for a Glu at the +1 position as it had been hypothesized previously. A triple mutant (Lys beta D3Ala, Asp beta C8Ala, AspCD2Ala) has reduced binding affinity compared to the double (Asp beta C8Ala, AspCD2Ala) mutant, yet binds the pYEEI peptide as well as the wild-type Src SH2 domain. The structural basis for such high affinity interaction was investigated crystallographically by determining the structure of the triple (Lys beta D3Ala, Asp beta C8Ala, AspCD2Ala) mutant bound to the octapeptide PQpYEEIPI (where pY indicates a phosphotyrosine). This structure reveals for the first time contacts between the SH2 domain and the -1 and -2 positions of the peptide (i.e. the two residues N-terminal to pY). Thus, unexpectedly, mutations in the +1 binding region affect binding of other regions of the peptide. Such additional contacts may account for the high affinity interaction of the triple mutant for the pYEEI-containing peptide. PMID- 11851340 TI - The SH3 domain of nebulin binds selectively to type II peptides: theoretical prediction and experimental validation. AB - Nebulin, a giant modular protein from muscle, is thought to act as a molecular ruler in sarcomere assembly. The C terminus of nebulin, located in the sarcomere Z-disk, comprises an SH3 domain, a module well known for its role in protein/protein interactions. SH3 domains are known to recognize proline-rich ligands, which have been classified as type I or type II, depending on their relative orientation with respect to the SH3 domain in the complex formed. Type I ligands are bound with their N terminus at the RT loop of the SH3 domain, while type II ligands are bound with their C terminus at the RT loop. Many SH3 domains can bind peptides of either class. Despite the potential importance of the SH3 domain for the function of nebulin as an integral part of a complex network of interactions, no in vivo partner has been identified so far. We have adopted an integrated approach, which combines bioinformatic tools with experimental validation to identify possible partners of nebulin SH3. Using the program SPOT, we performed an exhaustive screening of the muscle sequence databases. This search identified a number of potential nebulin SH3 partners, which were then tested experimentally for their binding affinity. Synthetic peptides were studied by both fluorescence and NMR spectroscopy. Our results show that nebulin SH3 domain binds selectively to type II peptides. The affinity for a type II peptide, 12 residues long, spanning the sequence of a stretch of titin known to colocalise with nebulin in the Z-disk is in the submicromolar range (0.7 microM). This affinity is among the highest found for SH3/peptide complexes, suggesting that the identified stretch could have significance in vivo. The strategy outlined here is of more general applicability and may provide a valuable tool to identify potential partners of SH3 domains and of other peptide-binding modules. PMID- 11851341 TI - The solution structure and DNA-binding properties of the cold-shock domain of the human Y-box protein YB-1. AB - The human Y-box protein 1 (YB-1) is a member of the Y-box protein family, a class of proteins involved in transcriptional and translational regulation of a wide range of genes. Here, we report the solution structure of the cold-shock domain (CSD) of YB-1, which is thought to be responsible for nucleic acid binding. It is the first structure solved of a eukaryotic member of the cold-shock protein family and consists of a closed five-stranded anti-parallel beta-barrel capped by a long flexible loop. The structure of CSD is similar to the OB-fold and a comparison with bacterial cold-shock proteins shows that its structural properties are conserved from bacteria to man. Our data suggest the presence of a DNA-binding site consisting of a patch of positively charged and aromatic residues on the surface of the beta-barrel. Further, it is shown that CSD, which has a preference for binding single-stranded pyrimidine-rich sequences, binds weakly and hardly specifically to DNA. Binding affinities reported for intact YB 1 indicate that domains other than the CSD play a role in DNA binding of YB-1. PMID- 11851342 TI - Comparison of the folding processes of T. thermophilus and E. coli ribonucleases H. AB - In order to examine how the stabilization of thermophilic proteins affects their folding, we have characterized the folding process of Thermus thermophilus ribonuclease H using circular dichroism, fluorescence, and pulse-labeling hydrogen exchange. Like its homolog from Escherichia coli, this thermophilic protein populates a partially folded kinetic intermediate within the first few milliseconds of folding. The structure of this intermediate is similar to that of E.coli RNase H and corresponds remarkably well to a partially folded form that is populated at low levels in the native state of the protein. Proline isomerization appears to partly limit the folding of the thermophilic but not the mesophilic protein. Lastly, unlike other thermophilic proteins, which unfold much more slowly than their mesophilic counterparts, T.thermophilus RNase H folds and unfolds with overall rates similar to those of E.coli RNase H. PMID- 11851343 TI - Wavelet transforms for the characterization and detection of repeating motifs. AB - The role of repeating motifs in protein structures is thought to be as modular building blocks which allow an economic way of constructing complex proteins. In this work novel wavelet transform analysis techniques are used to detect and characterize repeating motifs in protein sequence and structure data, where the Kyte-Doolittle hydrophobicity scale (Eta Phi) and relative accessible surface area (rASA) data provide residue information about the protein sequence and structure, respectively. We analyze a variety of repeating protein motifs, TIM barrels, propellor blades, coiled coils and leucine-rich repeat structures. Detection and characterization of these motifs is performed using techniques based on the continuous wavelet transform (CWT). Results indicate that the wavelet transform techniques developed herein are a promising approach for the detection and characterization of repeating motifs for both structural and in some instances sequence data. PMID- 11851344 TI - Multiple site-specific infrared dichroism of CD3-zeta, a transmembrane helix bundle. AB - The structure of the transmembrane domain of CD3-zeta a component of the T-cell receptor involved in signal transduction, has been studied in its native state (a lipid bilayer) by multiple site-specific infrared dichroism. For the first time, the transmembrane domain has been labelled at multiple positions along the sequence, representing a total of 11 samples, each labelled at a different residue with an isotopically modified carbonyl group, (13)C [double bond] (18)O. A strategy is outlined that, based on the above data, can yield the rotational orientation and the local helix tilt for each labelled residue, giving a detailed description of helix geometry. The results obtained indicate that the transmembrane segment is in an alpha-helical conformation throughout, with an average helix tilt of 12 degrees. The N-terminal side of the helix is more tilted than the C-terminal. In an accompanying paper we describe the implementation of the infrared data in a model-building study of the CD3-zeta transmembrane complex. The model obtained is entirely consistent with results based on evolutionary conservation data. Taken together, this study represents the first step towards elucidation of the backbone structure of a transmembrane alpha helical bundle by infrared spectroscopy. PMID- 11851345 TI - Convergence of experimental, computational and evolutionary approaches predicts the presence of a tetrameric form for CD3-zeta. AB - Experimental results using multiple site-specific infrared dichroism have shown that, when reconstituted into lipid bilayers, the orientation of the transmembrane domain of CD3-zeta is not compatible with a dimeric right-handed model reported previously. This model, obtained using a computational approach that uses evolutionary data, is in agreement with mutagenesis data and homology modelling. This suggested that, in our experimental conditions, the oligomeric state of CD3-zeta may not be dimeric. We have explored this possibility by performing global searching molecular dynamics simulations assuming different homo-oligomeric sizes (from 2 to 6). In these simulations, the helix tilt was restrained to the average helix tilt obtained experimentally, 12 degrees. Only a left-handed tetrameric model was compatible with the experimentally observed tilt and rotational orientation of the helix, and was also the lowest-energy model amongst the candidate structures obtained. Furthermore, simulations performed using close homologues demonstrate that this model is compatible with evolutionary conservation data. Finally, the pattern of residue conservation in the zeta family of proteins strongly argues in favour of the presence of a left handed hetero-oligomer with an orientation compatible with the tetramer we present. These results show that both the known dimeric and the so far undetected tetrameric form may be of functional importance in the cell. PMID- 11851346 TI - Peptide investigations of pairwise interactions in the collagen triple-helix. AB - Pairwise interactions have been studied for the major secondary structures in proteins. The present work extends the characterization of interactions between side-chains to the context of a collagen triple-helix. In this study, the most frequent Gly-X-Y tripeptide sequences in collagen are characterized in terms of interchain interactions between non-imino acid X and Y residues, through the use of host-guest peptides and statistical frequency analysis. Stabilities predicted on the basis of additivity show good agreement with experimental values for almost half of the peptides, indicating a lack of interaction. A small number of peptides have a stability lower than predicted, while a larger number are more stable than expected. Of all triplets containing residues of opposite charge, only Gly-Lys-Asp and Gly-Arg-Asp exhibit stabilizing electrostatic interactions, and these pairs are found together preferentially in collagens. Repulsion of like charges is observed in Gly-Arg-Lys, Gly-Lys-Arg, and Gly-Glu-Asp sequences, and a small degree of hydrophobic stabilization was observed for the Gly-Leu-Leu guest triplet. The data reported here help clarify basic principles of triple-helix stability. In addition, the experimentally determined stabilities of the tripeptide units found most frequently in collagens constitute a database useful for predicting triple-helix stability in peptides, collagens and other triple helix-containing proteins. PMID- 11851347 TI - Hybridization of alpha class subunits generating a functional glutathione transferase A1-4 heterodimer. AB - Within the Alpha class of the mammalian glutathione transferases two variants of subunit interfaces exist. One is conserved among the A4 subunits, whereas the second one is found in all other members of the Alpha class. The ability of the two Alpha class subunit interfaces to adopt a functional heterodimeric structure has been investigated here.The heterodimer GST A1-4 was obtained by co-expression of the two human Alpha class subunits A1 and A4 in Escherichia coli. A histidine tail was added to the N terminus of the A1 subunit to facilitate the purification of the heterodimer. The heterodimer was formed in a small proportion implying that the efficiency of the hybridization between subunit A1 and A4 is less than the propensity for homodimer formation. The hybrid enzyme was stable at low temperatures, but the two subunits dissociated and reassociated into homodimers at 40 degrees C. Three different substrates were used for subunit-selective kinetic characterization of the GST A1-4 heterodimer: 1-chloro-2,4 dinitrobenzene, nonenal and Delta(5)-androstene-3,17-dione. Both subunit A1 and subunit A4 were active in GST A1-4, but the specific activities and k(cat) values were lower than the average values of the two parental isoenzymes. However, at high temperatures the subunits of the hybrid enzyme dissociated and formed homodimers, and the activities increased to expected values. Hence, the low activities of the individual subunits in the heterodimer were reversible. The non additive kinetic properties of the subunits in the heterodimer therefore highlight the importance of fine-tuned subunit interactions for optimal catalytic efficiency of GST A1-1 and GST A4-4. PMID- 11851348 TI - Cellular indigestion: chaperones head to the cytoskeleton. PMID- 11851349 TI - Mending broken hearts one cell at a time. PMID- 11851350 TI - Myocyte growth and cardiac repair. AB - Introduced several decades ago, the dogma persists that ventricular myocytes are terminally differentiated cells and cardiac repair by myocyte regeneration is completely inhibited shortly after birth. On the basis that cardiac myocytes are unable to divide in the adult heart, myocyte growth under physiologic and pathologic conditions is believed to be restricted to cellular hypertrophy. Evidence is presented to indicate that this old paradigm has to be changed to include myocyte replication as a significant component of the cellular processes of ventricular remodeling. Importantly, myocyte death, apoptotic and necrotic in nature, has to be regarded as an additional critical variable of the multifactorial events implicated in the alterations of cardiac anatomy and myocardial structure of the decompensated heart. Methodologies are currently available to recognize and measure quantitatively the contribution of myocyte size, number and death to the adaptation of the overloaded heart and its progression to cardiac failure. PMID- 11851351 TI - Survival and development of neonatal rat cardiomyocytes transplanted into adult myocardium. AB - Transplantation of neonatal cardiomyocytes is a novel approach for the treatment of heart failure and myocardial infarction, but quantitative information on long term cell survival and development is limited. Male donor cardiomyocytes were isolated from neonatal Fischer 344 rats (1-2 days), purified, and injected into the left ventricular wall of female syngeneic adult rats. One hour to 12 weeks later, genomic DNA was isolated from recipient hearts. The amount of male DNA per sample was determined by quantitative real-time TaqMan PCR of the male-specific Sry gene. Transplanted cell survival was 57 +/- 9% at 0-1 h, 24 +/- 6% at 24 h, 28 +/- 11% at 7 days, 27 +/- 3% at 14 days, 23 +/- 8% at 4 weeks and 15 +/- 3% at 12 weeks. The caspase inhibitor AcYVADcmk failed to improve transplanted cell survival at 24 h, suggesting that apoptosis did not play a major role in cell loss. Histology revealed that transplanted cells became more elongated over time, developed cross-striations, and that their nuclei increased in size. However, at 12 weeks, transplanted cells and their nuclei were still smaller than those of host myocardium. We established a quantitative survival profile for neonatal cardiomyocytes transplanted into normal adult myocardium. There was significant loss of cells within 24 h, but 15% of transplanted cells survived 12 weeks. Those cells that did survive underwent differentiation and developed visible sarcomeres, suggesting a potential contribution toward ventricular function. PMID- 11851352 TI - Translocation of small heat shock proteins to the actin cytoskeleton upon proteasomal inhibition. AB - The role of small heat shock proteins (sHsps) as molecular chaperones is still poorly understood. We therefore investigated the effect of proteasomal inhibition on sHsps in the rat cardiac myoblast cell line H9c2. Proteasomes are responsible for controlled degradation of intracellular proteins. Inhibition of their activities leads to accumulation of unfolded proteins, which can form insoluble "aggresomes" together with proteasomes and heat shock proteins Hsp70 and Hsp90. We here report that upon proteasome inhibition, alpha B-crystallin and Hsp25 translocate from the detergent-soluble cytosolic fraction to the detergent insoluble nuclear/cytoskeletal fraction. Although phosphorylation of both alpha B crystallin and Hsp25 is induced, this does not seem to be essential for the translocation. Immunocytochemistry revealed that alpha B-crystallin and Hsp25, which show a diffuse cytoplasmic staining in unstressed H9c2 cells, colocalize with F-actin upon proteasomal inhibition. After transfection in H9c2 cells, other sHsps (alpha A-crystallin, Hsp20, HspB2 and HspB3) showed similar translocation to the actin cytoskeleton. The redistribution of sHsps upon proteasomal inhibition may reflect a mechanism by which cells are protected from damaged intracellular proteins by sequestering them on the cytoskeleton. PMID- 11851353 TI - Antioxidant and lysosomotropic properties of acridine-propranolol: protection against oxidative endothelial cell injury. AB - The antioxidant and lysosomotropic properties of a fluorescent analogue of propranolol, 9-amino-acridine-propranolol (9-AAP) were compared to those of propranolol. Using isolated microsomal membranes exposed to a superoxide and hydroxyl radical generating system, 9-AAP was found to be at least 10-fold more potent than propranolol (and about 50% as potent as vitamin E) in inhibiting lipid peroxidation. In cultured endothelial cells, 9-AAP afforded moderate protective effect against acute loss of glutathione but potent cytoprotective activity against free radical-mediated loss of viability/survival. Intracellular localization of 9-AAP was examined by fluorescent microscopy and compared with two known fluorescent lysosomal markers: acridine orange and Lysosensor. All three agents appeared to localize to similar peri-nuclear vesicles, presumably lysosomes or pre-lysosomes. Lysosensor fluorescence was not observable in the presence of 9-AAP, foreclosing the possibility of a direct dual labeling experiment. We employed the pH sensitivity of acridine orange to determine if it labels the same vesicles as 9-AAP. When the endothelial cells were preloaded with acridine orange, washed and resuspended in buffer containing 9-AAP, the dark orange-labeled vesicles observed with acridine orange alone became increasingly lighter with time. Since the fluorescence of acridine orange is altered by pH change, this spectral shift in fluorescence emission is consistent with the indication that added propranolol (or the analog) leads to lysosomal alkalization. In conclusion, 9-AAP is both a strong antioxidant and a lysosomotropic agent that is remarkably insensitive to photobleaching. These properties may contribute to the enhanced endothelial cytoprotective effects against free radical-induced injury. PMID- 11851354 TI - Role of EGF Receptor and Pyk2 in endothelin-1-induced ERK activation in rat cardiomyocytes. AB - G protein-coupled receptor (GPCR)-evoked signal transduction pathways leading to the activation of extracellular signal-regulated kinases (ERK) are quite different among cell types. In cardiomyocytes, much attention has been focused on the activation of protein kinase C (PKC) or mobilization of intracellular Ca(2+) ([Ca(2+)](i)), however, the contributions of tyrosine kinases are controversial. In the present study, we characterized the signaling pathways involving tyrosine kinases, Pyk2 and epidermal growth factor receptor (EGFR), and their contribution to ERK activation in cultured cardiomyocytes. We initially investigated the potential involvement of [Ca(2+)](i) and PKC on the activation of these kinases in endothelin-stimulated cardiomyocytes. Interestingly, activation of Pyk2 was abrogated by chelating [Ca(2+)](i) or by downregulation of PKC, whereas transactivation of EGFR was solely dependent on PKC. By using a compound that selectively interferes with EGFR (AG1478), c-Src (PP1), or disrupts actin cytoskeleton (cytochalasin D), we demonstrated that cytochalasin D completely inhibited the activation of Pyk2, but not that of EGFR, whereas AG1478 did not inhibit the activation of Pyk2, indicating that transactivation of EGFR and signaling pathways involving Pyk2 were distinct pathways. Furthermore, activation of ERK and Shc, and c- fos gene expression were significantly inhibited by AG1478 but not by cytochalasin D or PP1. Overexpression of deletion mutant of EGFR attenuated the activation of ERK. These facts demonstrated the existence of two distinct tyrosine kinase pathways requiring Pyk2 or EGFR downstream from GPCR in cardiomyocytes. EGFR was Ca(2+)-independently activated and predominantly contributed to Shc/ERK/c- fos activation, while Pyk2 or c-Src contributed less to it. PMID- 11851355 TI - Effect of ramipril and furosemide treatment on interstitial remodeling in post infarction heart failure rat hearts. AB - Extracellular matrix (ECM) remodeling and increased matrix metalloproteinase (MMP) expression and activity have been observed to be relevant in the development of heart failure (HF). We examined the effects of ramipril alone or with furosemide on ECM in a heart failure model. HF was induced by occlusion of the left coronary artery in spontaneously hypertensive rats (SHR). Rats were assigned to placebo (n=9), ramipril 1 mg/kg/day (n=11), furosemide 2 x 2 mg/kg/day (n=7) or both (1 mg/kg/day + 2 x 2 mg/kg/day n=8). LV-function, collagen content, MMP/TIMP (tissue inhibitor of matrix metalloproteinases) protein- and mRNA-expression were examined in non-infarcted LV tissue. MMP-2/TIMP 4 ratio was increased in HF. Ramipril reduced MMP-2 expression (active form), collagen type I mRNA expression and content and increased TIMP-4 levels associated with decreased left ventricular end diastolic pressure (LVEDP), mortality rate and increased LV pressure (LVP). Combination therapy with furosemide is less efficient with regard to collagen content and MMP-2 (active form) reduction but did not worsen beneficial effects of ramipril on LV function and mortality rate. Furosemide alone had no effect on MMP-2 (active form) expression, collagen content, LV function and mortality rate. Prevention of LV dilatation by ramipril was associated with decreased gelatinolytic activity and increased MMP-inhibition in heart failure SHR. Furthermore, ramipril reduced fibrosis by enhanced interstitial collagenase expression. Furosemide did not show the beneficial effects of ramipril on ECM remodeling but did not worsen LV function. Positive effects of furosemide treatment alone on LV remodeling and function were not observed. PMID- 11851356 TI - Apoptosis and post-infarction left ventricular remodeling. AB - Apoptosis is a common pathological feature in acute myocardial infarction (AMI), however, its role in the later phases (>10 days) of AMI and in post-infarction left ventricular remodeling has not been characterized. The aim of the study was to identify signs of ongoing cell apoptosis late post AMI. Sixteen hearts were collected at autopsy from subjects 12 to 62 days after the onset of AMI. In situ end-labeling of DNA fragmentation (TUNEL) and co-staining with caspase-3 were performed. Double-positive cells were defined as apoptotic and the apoptotic rate was calculated. Values are expressed as median and interquartile range. Co stainings with muscle-actin, splicing factor (SC35), PCNA, bax and bcl-2 were also performed. Apoptotic rates at site of infarction [25.4% (17.0-28.4%)] were significantly higher v those at remote regions [0.7% (0.5-0.8%) P<0.001] and significantly correlated to left ventricular longitudinal and transverse diameters [ r = +0.70 (P=0.016) and r = +0.63 (P=0.004) respectively]. Moreover, in subjects with persistently occluded infarct-related artery (14 cases) there was a significantly higher apoptotic rate at the site of infarction compared to those (2 cases) with patent artery [26.0% (21.9-28.5%) v 4.5% (0.6% and 8,4%);P=0.033]. A significantly greater bax immuno-reactivity close to the infarction v remote areas was found (P<0.001). High grade apoptosis is present at sites of infarction in the later phases post AMI. This is more evident if the infarct-related artery is persistently occluded and signs of ventricular remodeling are present. These data may provide an explanation of progressive late left ventricular dysfunction. PMID- 11851357 TI - Cardiac structure and function in young and senescent mice heterozygous for a connexin43 null mutation. AB - Downregulation of connexin43 (Cx43) in the failing heart has been implicated not only in arrhythmogenesis but in contractile dysfunction as well. Cx43-deficient mice exhibit reduced baseline conduction velocity and increased arrhythmias in response to ischemia. However, it is not known whether Cx43-deficient mice have any abnormalities in contractile function or, furthermore, whether cardiac dysfunction may be manifested in Cx43-deficient mice with advancing age. Therefore, we analyzed echocardiographic images from young and senescent Cx43 deficient C57BL/6Jx129 mice compared to wild-type littermate controls. Only a few, modest genotype-related differences were observed. LV wall thickness during systole and % fractional shortening were diminished by 8-10% in Cx43-deficient v wild-type mice. Aging alone had a greater effect on cardiac structure and function. LV mass and relative wall thickness were significantly increased in senescent v young mice independent of genotype. Percent fractional shortening and LV internal chamber dimension were significantly reduced in senescent v young mice. Thus, aging in mice, as in humans, is associated with concentric remodeling, mild systolic dysfunction and fibrosis. Although diminished Cx43 expression could contribute to contractile dysfunction in patients with advanced heart failure, genetic deficiency in Cx43 does not appear significantly to alter cardiac structure or function even in aged mice. PMID- 11851358 TI - Alpha-adrenergic receptor stimulation produces late preconditioning through inducible nitric oxide synthase in mouse heart. AB - Inducible nitric oxide synthase (iNOS) mediates late preconditioning (PC) induced by ischemia and pharmacological agents. Since alpha -adrenoceptor (alpha -AR) stimulation is one of the key triggers of PC, we hypothesized that activation of this receptor may induce delayed cardioprotective effect via iNOS-sensitive mechanisms. Adult male ICR mice were treated i.p. with either vehicle/inhibitors or phenylephrine (10 mg/kg) and subjected to 30 min of global ischemia and 30 min reperfusion in Langendorff mode 24 h later. 5-Methyl-urapidil (3 mg/kg) and chloroethylclonidine (3 mg/kg) were injected 15 min prior to phenylephrine to block alpha -AR(1A) and alpha -AR(1B) receptors respectively. S-Methylisothiourea (3 mg/kg), an iNOS inhibitor, was given 60 min prior to ischemia in phenylephrine pretreated mice. Preischemic NO(x) was measured using a chemoluminescence reaction. Phenylephrine treatment reduced infarct size from 31.10 +/- 0.79% (vehicle) to 14.24 +/- 0.84% (P<0.001). Chloroethylclonidine blocked the effect of phenylephrine (infarct size 31.31 +/- 1.69%) but 5-methyl-urapidil (17.72 +/- 1.25%) did not. Phenylephrine-induced delayed cardioprotection was abolished by S methylisothiourea and absent in iNOS knockout mice. Baseline NO(x) content was significantly increased in phenylephrine and 5-methyl-urapidil+phenylephrine treated hearts, but remained at baseline levels in hearts treated with chloroethylclonidine, 5-methyl-urapidil or S-methylisothiourea. Western blot analysis revealed a 1.8-fold increase in iNOS with phenylephrine, which was inhibited by chloroethylclonidine but not by 5-methyl-urapidil. We conclude that phenylephrine-induced delayed PC is mediated by selective activation of alpha AR(1B). Enhanced iNOS expression concomitant with increased NO synthesis, as well as pharmacological blockade and absence of cardioprotection in iNOS knockout mice suggests an essential role of NO in phenylephrine triggered late PC. PMID- 11851359 TI - Effects of components of ischemia on the Kv4.3 current stably expressed in Chinese hamster ovary cells. AB - We investigated the effects of three components of ischemia: external acidosis (pH=6.0), extracellular hyperkalemia ([K(+)]=20 mmol/l), and resting membrane depolarization to -60 mV, on Kv4.3 current stably expressed in Chinese Hamster Ovary cells. We used single electrode whole cell patch clamp techniques to study changes in the current elicited. External acidosis caused a positive shift in the steady state activation curve from -13.4 +/- 2.1 mV to -3.3 +/- 1.5 mV (n=8, P=0.004) and the steady state inactivation curve from -56.5 +/- 0.4 mV to -46.7 +/- 0.5 mV (n=14, P<0.0001). Acidosis also caused an acceleration of recovery from inactivation with the t(1/2) decreasing from 306 ms (95% CI 287-327 ms) to 194 ms (95% CI 182-207 ms), (n=14, P<0.05). Hyperkalemia did not affect any of these parameters. Combined acidosis and hyperkalemia produced effects similar to those seen with acidosis. Changing the holding potential from -90 mV to -60 mV with test potentials of +5 and +85 mV decreased the peak currents by 34.1% and 32.4% respectively (n=14). However, in the presence of external acidosis the decrease in peak currents induced by changing the holding potential was less marked. In acidotic bath the peak current at -60 mV was reduced by only 13.6% at a test potential of +5 mV and 12.3% at a test potential of +85 mV (n=14). Taken together our data suggest that the membrane depolarization and changes in pH which occur under ischemic conditions would be accompanied by relative preservation of Kv4.3 currents and provide a molecular basis for the observation of preserved epicardial I(to) and epicardial action potential duration (APD) shortening in ischemia. PMID- 11851360 TI - Chemokine expression in myocardial ischemia: MIP-2 dependent MCP-1 expression protects cardiomyocytes from cell death. AB - Chemokines are small molecular weight proteins that play important roles in inflammation. Originally described as chemotactic cytokines, chemokines stimulate the influx of leukocytes into specific tissue compartments. These molecules also modulate gene expression in both infiltrating and resident cells to mediate a vast array of cellular functions, and their importance in disease processes has been well documented. This study examined the expression of chemokines during myocardial ischemia and established a pathway by which two, MIP-2 and JE/MCP-1, modulate cardiac myocyte viability during this process. To focus on the direct effects of chemokines on these cells, a mouse model of ischemia without reperfusion was used. The expression of chemokines and chemokine receptors was induced in the left ventricular free wall as early as 1 h post-ischemia, with the most significant increases in MIP-2 (CXCL2) and JE/MCP-1 (CCL2). Expression of their respective receptors, CXCR2 and CCR2, was also induced. Similar changes in gene expression occurred at the mRNA and protein levels in isolated neonatal mouse cardiac myocytes stimulated by hypoxia. Antibody to MIP-2 inhibited hypoxia induced JE/MCP-1 expression, demonstrating that MIP-2 is critical for this event. Moreover, in vivo intramyocardial injection of either an adenovirus expressing MIP-2 or the recombinant protein itself was sufficient to upregulate JE/MCP-1 production even in the absence of ischemia. Thus, MIP-2 regulates JE/MCP-1 expression both in cell culture and in vivo. Furthermore, JE/MCP-1 markedly decreased hypoxia-induced cell death in cultured cardiac myocytes. Thus, JE/MCP-1 appears to mediate an unanticipated survival pathway in target cardiac myocytes themselves. These findings indicate an important role for MIP-2 and JE/MCP-1 in regulating the response of cardiac myocytes to myocardial ischemia. PMID- 11851361 TI - Alterations of the circadian clock in the heart by streptozotocin-induced diabetes. AB - The heart, like other organs, possesses an internal circadian clock. These clocks provide the selective advantage of anticipation, enabling the organ to prepare for a given stimulus, thereby optimizing the appropriate response. The heart in diabetes is associated with alterations in morphology, gene expression, metabolism and contractile performance. The present study investigated whether diabetes also alters the circadian clock in the heart. Insulin-dependent diabetes mellitus was induced in rats by treatment with streptozotocin (STZ; 65 mg/kg). STZ increased humoral (glucose and non-esterified fatty acids) and heart gene expression (myosin heavy chain beta, pyruvate dehydrogenase kinase 4 and uncoupling protein 3) markers of diabetes. The circadian patterns of gene expression of seven components of the mammalian clock (bmal1, clock, cry1, cry2, per1, per2 and per3), as well as three clock output genes (dbp, hlf and tef), were compared in hearts isolated from control and STZ-induced diabetic rats. All components of the clock investigated possessed circadian rhythms of gene expression. In the hearts isolated from STZ-induced diabetic rats, the phases of these circadian rhythms were altered (approximately 3 h early) compared to those observed for control hearts. The clock in the heart has therefore lost normal synchronization with its environment during diabetes. Whether this loss of synchronization plays a role in the development of contractile dysfunction of the heart in diabetes remains to be determined. PMID- 11851362 TI - Involvement of reactive oxygen species-mediated NF-kappa B activation in TNF alpha-induced cardiomyocyte hypertrophy. AB - We examined the intracellular signaling mechanism for tumor necrosis factor-alpha (TNF-alpha)-induced cardiac hypertrophy in isolated rat neonatal cardiomyocytes. TNF-alpha enhanced the expression of a kappa B-dependent reporter gene construct in a dose-dependent manner, which was transiently transfected in cardiomyocytes. Electrophoretic mobility shift assay demonstrated that TNF-alpha induced nuclear factor- kappa B (NF-kappa B)-specific DNA binding. Cultured cardiomyocytes were infected with a recombinant adenoviral vector expressing a degradation-resistant mutant of I kappa B alpha (AdI kappa B alpha 32/36A). The I kappa B alpha mutant suppressed NF-kappa B activation induced by TNF- alpha. In cardiomyocytes infected with AdI kappa B alpha 32/36A, TNF-alpha-induced hypertrophic responses, including increases in cell size, protein synthesis and atrial natriuretic factor production and enhancement of sarcomeric organization, were remarkably attenuated compared to the cells infected with an adenovirus expressing bacterial beta galactosidase. Using a reactive oxygen species (ROS)-sensitive fluorescent dye, 2', 7'-dichlorofluorescin, we observed an increase in fluorescent signal in cardiomyocytes over time, upon addition of TNF-alpha. Preincubation of n-acetyl cysteine (NAC), an antioxidant, prior to TNF-alpha treatment, abolished TNF-alpha -induced ROS generation. NAC abolished TNF-alpha-induced NF-kappa B activation and hypertrophic responses. These findings indicated that TNF-alpha-induced cardiomyocyte hypertrophy is mediated through NF-kappa B activation via the generation of ROS. PMID- 11851363 TI - Skeletal muscle stem cells do not transdifferentiate into cardiomyocytes after cardiac grafting. AB - Skeletal muscle cell-derived grafts in the heart may benefit myocardial performance after infarction. Several studies have suggested that skeletal muscle stem cells (satellite cells) from adult muscle undergo transdifferentiation into cardiomyocytes after grafting into the heart, but expression of cardiac markers in graft cells has not been rigorously confirmed. To determine the fate of satellite cell-derived grafts in the heart, adult rat satellite cells were tagged in vitro with bromodeoxyuridine (BrdU) and grafted into normal hearts of syngeneic rats. At 4 and 12 weeks the graft cells formed multinucleated, cross striated myofibers that expressed fast skeletal myosin heavy chain (MHC), thus indicating a mature skeletal muscle phenotype. Double staining for the BrdU tag and cardiac-specific markers was employed to identify transdifferentiation. Aside from four questionable cells, none of the 11 grafts examined expressed alpha-MHC, cardiac troponin I, or atrial natriuretic peptide. At 4 weeks, grafts expressed beta -MHC, a hallmark of slow twitch myofibers. By 12 weeks, however, the myofibers had atrophied and downregulated beta-MHC. Grafts never expressed the intercalated disk proteins N-cadherin or connexin43, hence electromechanical coupling did not occur. In conclusion, satellite cells differentiate into mature skeletal muscle and do not express cardiac-specific genes after grafting into the heart. Thus, transdifferentiation into cardiomyocytes did not occur. PMID- 11851364 TI - A mechanistic model of algal photoinhibition induced by photodamage to photosystem-II. AB - Photoinhibition is a central problem for the understanding of plasticity in photosynthesis vs. irradiance response. It effectively reduces the photosynthetic rate. In this contribution, we present a mechanistic model of algal photoinhibition induced by photodamage to photosystem-II. Photosystem-IIs (PSIIs) are assumed to exist in three states: open, closed and inhibited. Photosynthesis is closely associated with the transitions between the three states. The present model is defined by four parameters: effective cross section of PSII, number of PSIIs, turnover time of electron transfer chains and the ratio of rate constant of damage to that of repair of D1 proteins in PSIIs. It gives a photosynthetic response curve of phytoplankton to irradiance (PI-curve). Without photoinhibition, the PI-curve is in hyperbola with the first three parameters. The PI-curve with photoinhibition can be simplified to the same form as the hyperbola by replacing either the number of PSIIs with the number of functional PSIIs or the turnover time of electron transfer chains with the average turnover time. PMID- 11851365 TI - Kinetic model of BiP- and PDI-mediated protein folding and assembly. AB - A mechanism for heavy chain binding protein (BiP)- and protein disulfide isomerase (PDI)- mediated protein folding and assembly has been proposed. It considers BiP chaperoning action and PDI catalytic activity. A kinetic model has been developed based on the proposed mechanism. The model was used for quantifying the influence of polypeptide concentration and ratio, and the effect of BiP and PDI concentration on the kinetics of folding and assembly. An optimum value for polypeptide concentration that minimizes assembly times was found, and different kinetic behaviors were identified for polypeptide concentrations higher or lower than the optimum. Pulse-chase experiments and the dependence of assembly time on unassembled polypeptides ratio predicted by the model are similar to those found during in vitro and in vivo folding and assembly of antibodies and human chorionic gonadotropin (hCG), as well as bovine pancreatic trypsin inhibitor (BPTI) folding. The model also explains the increase in folding and assembly rates during overexpression of BiP and PDI. PMID- 11851366 TI - A Maxwell's Demon type of membrane transport: possibility for active transport by ABC-type transporters? AB - ATP-binding-cassette (or ABC)-type transporters constitute one of the largest family of membrane transporters in nature. Many of its members move substrates "actively", i.e. in an ATP-dependent manner against an electrochemical gradient. No consensus is available about the mechanism. Therefore, a novel class of transport mechanisms is proposed based on Maxwell's demon idea. This transport mechanism consists of a gated pore that selectively opens for substrates from one, but not the other side. Thermoenergy (Brownian motion) would suffice for substrate translocation across the membrane; energy for synchronizing gate opening with substrate arrival would come from ATP hydrolysis. Simulations demonstrate that such a mechanism would be thermodynamically and kinetically feasible. It exhibits "active", unidirectional transport, saturation, and other typical features of protein-catalysed reactions. It also shows pore behavior with charged substrates moving under the influence of electrical potentials. Its efficiency depends on a diffusion time constant of the substrate in solution that is slower than the transit time through the membrane, a situation that can realistically be achieved at millimolar or lower substrate concentrations. Features of the novel mechanism that differ significantly from P- or F-type ATPases are: (1) transport cannot be run in "reverse" to synthesize ATP even if sufficient energy is available in the gradient of the transported solute and (2) unidirectional and net substrate fluxes through the transporter diverge with increasing substrate concentration. PMID- 11851367 TI - Directionality of stripes formed by anisotropic reaction-diffusion models. AB - Turing mechanism explains the formation of striped patterns in a uniform field in which two substances interact locally and diffuse randomly. In a twin paper, to explain the directionality of stripes on fish skin in closely related species, we studied the effect of anisotropic diffusion of the two substances on the direction of stripes, in the cases in which both substances have high diffusivity in the same direction. In this paper, we study the direction of stripes in more general situations in which the diffusive direction may differ between the two substances. We derive a formula for the direction of stripes, based on a heuristic argument of unstable modes of deviation from the uniform steady state. We confirm the accuracy of the formula by computer simulations. When the diffusive direction is different between two substances, the directions of stripes in the spatial pattern change smoothly with the magnitude of anisotropy of two substances. When the diffusive direction of the two substances is the same, the stripes are formed either parallel or perpendicular to the common diffusive direction, depending on the relative magnitude of the anisotropy. The transition between these two phases occurs sharply. PMID- 11851368 TI - On a kinetic origin of heredity: minority control in a replicating system with mutually catalytic molecules. AB - As the first step in an investigation of the origin of genetic information, we study how some species of molecules are preserved over cell generations and play an important role in controlling the growth of a cell. We consider a model consisting of protocells. Each protocell contains two mutually catalysing molecule species (X and Y), each of which has catalytically active and inactive types. One of the species Y is assumed to have a slower synthesis speed. Through divisions of the protocells, the system reaches and remains in a state in which there are only a few active Y and almost no inactive Y molecules in most protocells, through the selection of very rare fluctuations. In this state, the active Y molecules are shown to control the behavior of the protocell. The minority molecule species act as the carrier of heredity, due to the relatively discrete nature of its population, in comparison with the majority species which behaves statistically in accordance with the law of large numbers. The minority controlled state may give rise to a selection pressure for mechanisms that ensure the transmission of the minority molecule. Once those mechanisms are in place, the minority molecule becomes the ideal storage device for information to be transmitted across generations, thus giving rise to "genetic information". The relevance of this minority controlled state to evolvability is also discussed. PMID- 11851369 TI - A model for the enhancement of fitness in cyanobacteria based on resonance of a circadian oscillator with the external light-dark cycle. AB - Fitness enhancement based on resonating circadian clocks has recently been demonstrated in cyanobacteria [Ouyang et al. (1998). Proc. Natl Acad. Sci. U.S.A.95, 8660-8664]. Thus, the competition between two cyanobacterial strains differing by the free-running period (FRP) of their circadian oscillations leads to the dominance of one or the other of the two strains, depending on the period of the external light-dark (LD) cycle. The successful strain is generally that which has an FRP closest to the period of the LD cycle. Of key importance for the resonance phenomenon are observations which indicate that the phase angle between the circadian oscillator and the LD cycle depends both on the latter cycle's length and on the FRP. We account for these experimental observations by means of a theoretical model which takes into account (i) cell growth, (ii) secretion of a putative cell growth inhibitor, and (iii) the existence of a cellular, light sensitive circadian oscillator controlling growth as well as inhibitor secretion. Building on a previous analysis in which the phase angle was considered as a freely adjustable parameter [Roussel et al. (2000). J. theor. Biol.205, 321-340], we incorporate into the model a light-sensitive version of the van der Pol oscillator to represent explicitly the cellular circadian oscillator. In this way, the model automatically generates a phase angle between the circadian oscillator and the LD cycle which depends on the characteristic FRP of the strain and varies continuously with the period of the LD cycle. The model provides an explanation for the results of competition experiments between strains of different FRPs subjected to entrainment by LD cycles of different periods. The model further shows how the dominance of one strain over another in LD cycles can be reconciled with the observation that two strains characterized by different FRPs nevertheless display the same growth kinetics in continuous light or in LD cycles when present alone in the medium. Theoretical predictions are made as to how the outcome of competition depends on the initial proportions and on the FRPs of the different strains. We also determine the effect of the photoperiod and extend the analysis to the case of a competition between three cyanobacterial strains. PMID- 11851370 TI - Stoichiometry, herbivory and competition for nutrients: simple models based on planktonic ecosystems. AB - Models are examined in which two prey species compete for two nutrient resources, and are preyed upon by a predator that recycles both nutrients. Two factors determine the effective relative supply of the nutrients, hence competitive outcomes: the external nutrient supply ratio, and the relative recycling of the two nutrients within the system. This second factor is governed by predator stoichiometry--its relative requirements for nutrients in its own biomass. A model with nutrient resources that are essential for the competing prey is detailed. Criteria are given to identify the limiting nutrient for a food chain of one competitor with the predator. Increased supply of this limiting nutrient increases predator density and concentration of this nutrient at equilibrium, while decreasing the concentration of a non-limiting nutrient. Changes in supply or recycling of a non-limiting nutrient affect only the concentration of that nutrient. Criteria for the invasion of a second prey competitor are presented. When different nutrients limit growth of the resident prey and the invader, increased supply or recycling of the invader's limiting nutrient assists invasion, while increased supply or recycling of the resident's limiting nutrient hinders invasion. If the same nutrient limits both resident and invader, then changes in supply and recycling have complex effects on invasion, depending on species properties. In a parameterized model of a planktonic ecosystem, green algae and cyanobacteria coexist over a wide range of nitrogen:phosphorus supply ratios, without predators. When the herbivore Daphnia is added, coexistence is eliminated or greatly restricted, and green algae dominate over a wide range of supply conditions, because the effective supply of P is greatly reduced as Daphnia rapidly recycles N. PMID- 11851371 TI - Insects could exploit UV-green contrast for Landmark navigation. AB - Illumination-invariant detection of landmark features is a prerequisite for landmark navigation in insects. It is suggested that a contrast mechanism involving the UV and green receptors of insect eyes could guarantee a robust separation between natural objects as foreground and sky as background. Using a sensor with a UV and a green channel that in their spectral characteristics are close to the corresponding insect photoreceptors, data of natural objects and sky were collected. The data show that the two classes can be separated by a fixed threshold in the UV-green color space, offering an advantage over a purely UV based separation that would require a dynamic threshold. Based on a numerical method, UV-green antagonism is shown to guarantee a more reliable discrimination than UV-blue antagonism. PMID- 11851372 TI - The dual role of CD4 T helper cells in the infection dynamics of HIV and their importance for vaccination. AB - Given the role of the CD4 T helper cells in the development of memory CTL precursors, it seems beneficial to boost the CD4 T helper response in the context of vaccination against the human immunodeficiency virus (HIV). However, CD4 T cells are also the preferred targets of infection by HIV. Here, we address the question as to whether it is advantageous to stimulate the CD4 T helper cell response, as this will increase the pool of potential target cells of infection. To do so we formulated a mathematical model describing the interactions between virus-infected cells, susceptible cells, HIV-specific CD4 helper T cells, and CTL precursor (CTLp) and effector cells (CTLe). The effect of increased initial CD4 helper and CTLp numbers on the outcome of infection, as well as the effect on viral set point of increased CD4 T helper growth rate, CTL responsiveness and the rate at which CTLp and CTLe are produced were studied. We found that only when the virus has a low basic reproductive number does the number of CTLp and CD4 T helper cells at the moment of infection influence the outcome of infection. In this situation, high initial T helper and CTL numbers can switch the outcome from full-blown infection to virus control. However, this holds for virus with infectivity in a limited range, and current estimates of virus infectivity suggest that it is higher. In that case, only a vaccination protocol that increases CTL responsiveness, ideally in combination with the rate of production of CD4 T helper cells, may offer a solution as it can reduce the viral set point considerably. If brought under a certain level, the viral population might be unable to replicate any further. However, changing these parameters of the immune response is only beneficial when infection is controlled by CTL in the long term. When a CD4 lymphoproliferative response is mounted but the CTL response is not maintained, increasing the CD4 T helper growth rate is deleterious. PMID- 11851373 TI - On the aperiodic locomotor behavior of Halobacterium salinarium under periodic light stimuli. AB - Two long time series of swimming intervals of a bacterium inverting its motion under periodic light pulses are analysed. The associated next-period plots reveal, through their filiform structure, that the underlying dynamics are low dimensional. Using recently described properties of such dynamics, a simple second-order black-box model for the swimming intervals is derived and validated. The model reinforces the conjecture that this bacterium is endowed with an oscillator controlling the switching of the flagellar motor. PMID- 11851374 TI - Errors drive the evolution of biological signalling to costly codes. AB - Reduction of costs in biological signalling seems an evolutionary advantage, but recent experiments have shown signalling codes shifted to signals of high cost with an underutilization of low-cost signals. Here I derive a theory for efficient signalling that includes both errors and costs as constraints and I show that errors in the efficient translation of biological states into signals can shift codes to higher costs, effectively performing a quality control. The statistical structure of signal usage is predicted to be of a generalized Boltzmann form that penalizes signals that are costly and sensitive to errors. This predicted distribution of signal usage against signal cost has two main features: an exponential tail required for cost efficiency and an underutilization of the low-cost signals required to protect the signalling quality from the errors. These predictions are shown to correspond quantitatively to the experiments in which gathering signal statistics is feasible as in visual cortex neurons. PMID- 11851375 TI - High sequence divergence in the 5' non-coding region of reference Coxsackie B and ECHO viral strains and clinical isolates revealed by restriction fragment length polymorphism analysis. AB - We report the restriction fragment length polymorphism (RFLP) patterns of a 440 bp-long 5' non-coding region (5' NCR) amplification target of all 34 reference Coxsackie B and ECHO (enteric cytopathic human orphan) enterovirus strains and a total of 42 serotypically pre-assigned clinical isolates, in order to afford meaningful comparisons among these patterns and those of polioviruses. The RFLP patterns of reference Coxsackie B strains differed from one another and from those of polio and ECHO reference enteroviruses except from Coxsackie B1 and B2, which, although they differed from one another, had identical RFLP patterns with ECHO 17 and 13, respectively. The 28 ECHO reference strains formed a more variable viral group including strains with RFLP patterns distinct from one another and from those of polio and Coxsackie B enteroviruses, and others with RFLP pattern identities common to other ECHO viruses and Coxsackie B1 and B2 but not polioviruses. The RFLP patterns of the clinical isolates and their corresponding serotypically assigned reference Coxsackie B and ECHO strains presented the most notable variations. The observed differences between serotype and genotype-dependent assignments within the 440-bp long 5' NCR target sequence of Coxsackie B and ECHO enteroviruses were in sharp contrast to the analogous situation with polioviruses. These findings support the specificity of the described method for clinical diagnostic genotyping of polioviruses and demonstrate that the 440-bp-long target sequence follows a different evolutionary process in polio and non-polio enteroviruses that is particularly prominent between reference non-polio strains and their serotypically assigned clinical isolates. PMID- 11851376 TI - Low-density lipoprotein receptor gene mutation analysis and clinical correlation in Belgian hypercholesterolaemics. AB - It is generally believed that patients with familial hypercholesterolaemia (FH) have a higher cardiovascular risk than hypercholesterolaemics without a defect in the low-density lipoprotein receptor (LDLR) gene. However, no conclusive evidence to support this view has yet been presented. We investigated this aspect in Belgian hyperlipidaemics as part of a comprehensive effort to determine the impact of FH in this population. DNA samples of 98 unrelated Belgian patients with a family history of autosomal dominant hypercholesterolaemia were screened for mutations in the LDLR gene, after exclusion of known mutations causing familial defective apolipoprotein B-100 (FDB). Eight of the 22 distinct LDLR gene mutations identified in 27 subjects have not previously been described in other populations. As expected, the mutation-positive patients had a significantly worse lipid profile than the mutation-negative subjects (p<0.05), but this did not correlate with clinical cardiovascular status. In conclusion, the presence of a mutation in the LDLR gene was not a reliable predictor of cardiovascular risk in the hyperlipidaemic subjects included in this study. However, it is possible that prolonged exposure to the high levels of LDL cholesterol in genetically proven FH patients will in future cause a higher incidence of coronary heart disease. Our data may reflect the genetic heterogeneity of inherited hypercholesterolaemia, recently shown to be caused by several major genes. PMID- 11851377 TI - PCR primers targeting the 16S rRNA gene for the specific detection of streptomycetes. AB - Streptomycetes are filamentous actinobacteria commonly found in soil and biotechnically important, but they also have adverse effects on human health. In this work, two primer pairs, StrepB/StrepE and StrepB/StrepF combined with Bst YI restriction endonuclease digestion, targeting the 16S rRNA gene of streptomycetes were designed. The specificity of the primers was determined by polymerase chain reaction (PCR) amplification from Streptomyces strains and near relatives. All streptomycetes tested positive and non-streptomycetes were not amplified except three strains that, however, gave Bst YI restriction endonuclease digestion results distinct from streptomycetes. Moreover, both primer pairs gave an amplification product of the expected size only when Streptomyces VTT E-99-1334 DNA was present in the template DNA mixture isolated from six bacterial and three fungal strains. The primers were further successfully used to amplify from DNA isolated from two soil and two building material samples. The 40 sequenced amplification products obtained with the primer pair StrepB/StrepE showed greater than 96.1% similarity to streptomycete 16S rRNA sequences. Seventy PCR amplification products obtained with the primers StrepB/StrepF were analysed by sequencing and restriction analysis. All 54 PCR products having >95.7% similarity to streptomycete sequences were cleaved with Bst YI. No false-positive results were achieved. Both primer sets proved to be specific for streptomycetes, and applicable for the detection of streptomycetes in environmental samples. PMID- 11851378 TI - Identification of predominant human and animal anaerobic intestinal bacterial species by terminal restriction fragment patterns (TRFPs): a rapid, PCR-based method. AB - Identification of predominant human and animal intestinal tract anaerobes by conventional methods is cumbersome, time-consuming and less sensitive as compared to molecular methods. We have developed a molecular technique to identify most of the abundant intestinal microflora by polyermase chain reaction (PCR) amplification of a 16S rRNA gene fragment using a pair of universal PCR primers. The forward PCR primer was labelled with 6-carboxyfluorescein amino hexy (6-FAM) fluorescent dye to detect the terminal fragment of the PCR products after digestion with restriction enzymes. The PCR products were purified and digested with restriction enzymes and were analysed by capillary electrophoresis using an automated DNA sequencer. The data was analysed with GeneScan software 2.1. Eleven bacteria (Eubacterium biforme, E. limosum, Peptostreptococcus productus, Lactobacillus acidophilus, Bacteroides thetaiotaomicron, B. vulgatus, B. distasonis, Clostridium clostridiiforme, C. leptum, C. perfringens and Escherichia coli) that are predominant in human and animal intestinal tract were successfully identified by this rapid molecular technique. This protocol is rapid, accurate, sensitive and capable of identifying multiple organisms in a single sample. PMID- 11851379 TI - Novel NPR1 polymorphic variants and its exclusion as a candidate gene for medullary cystic kidney disease (ADMCKD) type 1. AB - Autosomal dominant medullary cystic kidney disease (ADMCKD) is an adult-onset heterogeneous genetic nephropathy characterized by salt wasting and end-stage renal failure. The gene responsible for ADMCKD-1 was mapped on chromosome 1q21 and it is flanked proximally by marker D1S498 and distally by D1S2125, encompassing a region of approximately 8 cm. Within this region there are a large number of transcribed genes including NPR1 that encodes the atrial natriuretic peptide receptor 1. This receptor plays a crucial role in regulation of blood pressure by facilitating salt excretion. Based on its function we hypothesized this gene as a reasonable candidate for the MCKD1 locus. DNA mutation screening was performed on the entire NPR1 gene-coding sequence and some of the 5' prime UTR and 3'-UTR sequences. The samples investigated belonged to patients of five large ADMCKD-1 Cypriot families. The screening revealed two novel polymorphisms, one intragenic at amino acid position 939, which was occupied by either arginine or glutamine, and a second one located in the 3' prime-UTR, 29 nucleotides downstream of the NPR1 stop codon. The latter was a single nucleotide C insertion/deletion in a stretch of three or four Cs. No relationship was present between any allele of the two polymorphisms and the disease, as both alleles were observed in both affected and healthy subjects. In addition, no association was observed between the disease and another rare 8-bp deletion polymorphism at the 5' prime-UTR of NPR1 and the disease. Based on these findings it is unlikely that NPR1 is the same as the MCKD1 gene, although it is presently unknown whether it plays a disease modifying role. PMID- 11851380 TI - HyBeacon probes: a new tool for DNA sequence detection and allele discrimination. AB - Technologies that permit rapid investigation of DNA sequences, such as those containing single nucleotide polymorphisms (SNPs), are of great consequence to many sectors that perform molecular diagnostic analyses. We have developed a novel fluorescent oligonucleotide probe technology, termed HyBeacons, which provides a new homogeneous method for fluorescence-based sequence detection, allele discrimination and DNA quantification. Hybridization of HyBeacons to complementary DNA target sequences results in a measurable elevation of probe fluorescence emission. HyBeacon probes may be incorporated into real-time polymerase chain reaction (PCR) assays to detect the presence and monitor the accumulation of specific DNA sequences. Furthermore, closely related sequences differing by as little as a single nucleotide may be discriminated by measuring the melting temperatures (T(m)) of various probe/target duplexes and exploiting the differences in T(m) that exist between different duplexes. We demonstrate here that HyBeacon probes are efficient tools for rapid sequence analysis and that a single probe may be employed to reliably identify homozygous and heterozygous samples. Additional benefits exhibited by the HyBeacon technology derive from their simple mode of action, ease of design, relatively inexpensive synthesis and potential for multiplex analysis. PMID- 11851381 TI - Rapid and specific detection of the Mycobacterium tuberculosis complex using fluorogenic probes andreal-time PCR. AB - A rapid and sensitive strategy for the specific identification of Mycobacterium tuberculosis (TB) was designed and evaluated using crude mycobacterial lysates. The speed of real-time polymerase chain reaction (PCR) was combined with the sensitivity of fluorogenic probes to confirm the presence of mycobacteria as well as specifically identify the presence of members of the mycobacteria tuberculosis complex (MTC) in a single-tube assay. Oligonucleotides were designed to amplify the internal transcribed spacer (ITS) from several mycobacterial species. Specific fluorogenic probes were included in the PCR reaction for the identification of TB as well as Mycobacterium bovia and Mycobacterium africanum in bacterial lysates. The combination of TB-specific fluorogenic probes with real time PCR formed an approach determined to be fast (less than 40 min), sensitive (less than 800 copies of DNA) and reliable for the specific detection of the MTC. Our data demonstrate the use of real-time PCR and fluorogenic probes in a rapid and sensitive assay to distinguish members of the MTC from other mycobacterial species. PMID- 11851382 TI - Development of a multiplex PCR for the detection of Haplosporidium nelsoni, Haplosporidium costale and Perkinsus marinus in the eastern oyster (Crassostrea virginica, Gmelin, 1971). AB - Haplosporidium costale (SSO), Haplosporidium nelsoni (MSX) and Perkinsus marinus (Dermo) have caused oyster mortality on the North American east coast since the 1950s. Currently, the monitoring of oyster populations for these pathogens depends on histopathology for H. nelsoni, H. costale and the Ray/Mackin assay for P. marinus. In this study we describe the development and optimization of a multiplex polymerase chain reaction (MPCR) for the detection of H. nelsoni, H. costale and P. marinus. In addition, we determine its specificity and sensitivity. The MPCR clearly detects and differentiates the protozoan pathogens. There was no cross-reactivity between these species. The MPCR was able to detect DNA from H. nelsoni as low as 10 fg and P. marinus and H. costale as low as 1 pg. The MPCR allows for the detection of H. nelsoni, H. costale and P. marinus in a single PCR reaction. PMID- 11851383 TI - Identification of a single nucleotide polymorphism in the human alpha 2 delta 2 calcium channel subunit gene. PMID- 11851384 TI - C306A single nucleotide polymorphism in the human CEBPD gene that maps at 8p11.1 p11.2. PMID- 11851385 TI - Development of experimental and theoretical crystal lattice organic photochemistry: the quantitative cavity. Mechanistic and exploratory organic photochemistry. AB - The photochemistry of organic reactants in the crystalline state has a long history. What has been lacking is comprehensive theory defining what controls the course of these reactions which quite often afford products not obtainable in solution. Also lacking has been solid-state rearrangement chemistry with accompanying theory. In our research over the past two decades we have developed theoretical treatments of solid-state reactivity. This report describes the development of this research from its primitive beginnings to the present. PMID- 11851386 TI - Recognition in the minor groove of double-stranded DNA by microgonotropens. AB - Microgonotropens consist of a minor groove binding moiety functionalized with alkylamine side chains which are protonated and positively charged at neutral pH and capable of forming electrostatic bonds with the negatively charged phosphodiester backbone of DNA. We have investigated the interactions between microgonotropens and double-stranded DNA by a combination of fluorescence, ultraviolet, and nuclear magnetic resonance spectroscopy. Our work reveals why microgonotropens are such potent inhibitors of the binding of transcription factors to their DNA binding sites. PMID- 11851387 TI - Theoretical investigation of the mechanism of cis-trans regulation for the allylnickel(II)-catalyzed 1,4 polymerization of butadiene. AB - In this Account we summarize the recent progress in the computational modeling of the transition-metal-catalyzed 1,3-diene polymerization. We present a comprehensive and theoretically well-founded view of the cis-trans regulation mechanism of the intriguing C-C coupling for the allylnickel(II)-catalyzed 1,4 polymerization of butadiene. The crucial elementary reactions of the entire polymerization process were theoretically explored for typical trans-1,4- and cis 1,4-regulating catalysts. As a result, the catalytic structure-activity relationships are deduced, which are responsible for opening that reaction channel which yields trans-1,4 and cis-1,4 polymer units. PMID- 11851388 TI - Isotope effects in the study of phosphoryl and sulfuryl transfer reactions. AB - Phosphoryl and sulfuryl transfer reactions are essential biological processes. Multiple kinetic isotope effects have provided significant insights into the transition states of these reactions. The data are reviewed for the uncatalyzed reactions of phosphate and sulfate monoesters and for a number of enzymatic phosphoryl transfer reactions. Uncatalyzed phosphoryl and sulfuryl hydrolysis reactions are found to have very similar transition states. The phosphoryl transfer reaction catalyzed by protein-tyrosine phosphatases proceeds by a transition state very similar to that of the uncatalyzed reaction, but isotope effect data reveal an interesting interplay between the conserved arginine and enzyme dynamics involving general acid catalysis. PMID- 11851389 TI - Chemical and catalytic mechanisms of carboxyl transfer reactions in biotin dependent enzymes. AB - Biotin-dependent carboxylases catalyze a variety of carboxyl transfer reactions in a number of metabolic pathways and are found in all free-living organisms. They are large molecules which can comprise a single polypeptide chain with three domains or up to three subunits, each of which performs a particular part of the overall reaction. Biotin plays a central role in the action of these enzymes. In this Account we examine the current state of knowledge of the chemistry of catalysis and consider how the recent explosion of available protein sequence and structural information has assisted our understanding of the mechanisms of biotin dependent enzymes. PMID- 11851390 TI - Molecular engineering of semiconductor surfaces and devices. AB - Grafting organic molecules onto solid surfaces can transfer molecular properties to the solid. We describe how modifications of semiconductor or metal surfaces by molecules with systematically varying properties can lead to corresponding trends in the (electronic) properties of the resulting hybrid (molecule + solid) materials and devices made with them. Examples include molecule-controlled diodes and sensors, where the electrons need not to go through the molecules (action at a distance), suggesting a new approach to molecule-based electronics. PMID- 11851391 TI - Kinetics of chemical reactions on solid surfaces: deviations from conventional theory. AB - Isothermal kinetic measurements on elementary steps between species adsorbed on solid surfaces have highlighted significant deviations from conventional kinetic theories. In particular, it has been found that neighboring adsorbates modify the energetics of surface reaction in ways not explained by macroscopic kinetic models. The free energy barriers associated with transition state theory can be expanded to include coverage-dependent terms, but that does not account for local effects due to surface islanding. Changes in surface concentrations also lead to changes in adsorption geometries, and even to different adsorbate-surface bonding. Future Monte Carlo simulations and other theoretical approaches to describe surface kinetics need to include these factors. PMID- 11851393 TI - Mapping proximity within proteins using fluorescence spectroscopy. A study of T4 lysozyme showing that tryptophan residues quench bimane fluorescence. AB - We present a novel method for mapping proximity within proteins. The method exploits the quenching of the fluorescent label bimane by nearby Trp residues. In studies of T4 lysozyme we show that this effect appears to be distance dependent and orientation specific. Specifically, we show that a proximal Trp residue can reduce bimane fluorescence intensity by up to 500% and induce complicated fluorescence decay kinetics. Replacing the neighboring Trp residue with phenylalanine removes these spectral perturbations. The advantages of using the Trp quenching of bimane fluorescence for protein structural studies include the low amount of protein required and the substantial simplification of labeling strategies. We anticipate this method will prove suitable for a wide array of high-throughput protein studies such as protein folding, the detection of protein protein interactions, and, most importantly, the dynamic monitoring of conformational changes. PMID- 11851394 TI - Crystal structure of F65A/Y131C-methylimidazole carbonic anhydrase V reveals architectural features of an engineered proton shuttle. AB - The crystal structure of F65A/Y131C murine alpha-carbonic anhydrase V (CAV), covalently modified at cysteine residues with 4-chloromethylimidazole, is reported at 1.88 A resolution. This modification introduces a methylimidazole (MI) group at residue C131 in the active site with important consequences. F65A/Y131C-MI CAV exhibits an up to 3-fold enhancement of catalytic activity over that of wild-type CAV [Earnhardt, J. N., Wright, S. K., Qian, M., Tu, C., Laipis, P. J., Viola, R. E., and Silverman, D. N. (1999) Arch. Biochem. Biophys. 361, 264 270]. In this modified CAV variant, C131-MI acts as a proton shuttle, facilitating the deprotonation of a zinc-bound water molecule to regenerate the nucleophilic zinc-bound hydroxide ion. A network of three hydrogen-bonded water molecules, across which proton transfer likely proceeds, bridges the zinc-bound water molecule and the C131-MI imidazole group. The structure of F65A/Y131C-MI CAV is compared to structures of Y64H/F65A murine CAV, wild-type human alpha carbonic anhydrase II, and the gamma-carbonic anhydrase from Methanosarcina thermophilain an effort to outline common features of catalytic proton shuttles. PMID- 11851395 TI - Flavin recognition by an RNA aptamer targeted toward FAD. AB - Flavin adenine dinucleotide (FAD) is one of the primary cofactors in biological redox reactions. Designing cofactor-dependent redox ribozymes could benefit from studies of new RNA-cofactor complexes, as would our understanding of ribozyme evolution during an RNA World. We have therefore used the SELEX method to identify RNA aptamers that recognize FAD. Functional analysis of mutant aptamers, S1 nuclease probing, and comparative sequence analysis identified a simple, 45 nt helical structure with several internal bulges as the core-binding element. These aptamers recognize with high specificity the isoalloxazine nucleus of FAD but do not distinguish FAD from FADH(2), nor are they removed from an FAD resin with UMP (which shares a pattern of hydrogen bond donors and acceptors along one face). Thus, these aptamers are structurally and functionally distinct from previously identified FMN and riboflavin aptamers. Circular dichroism data suggest a conformational change in the RNA upon FAD binding. These aptamers require magnesium and are active across a wide pH range (4.5-8.9). Since general acid base catalysis plays a role in some flavin-dependent redox reaction mechanisms, these aptamers may be particularly well-suited to the design of new redox ribozymes. PMID- 11851396 TI - Interactions of RXR with coactivators are differentially mediated by helix 11 of the receptor's ligand binding domain. AB - RXR is a nuclear hormone receptor that is activated by the vitamin A metabolite 9 cis-retinoic acid. Previously, it was shown that, in the absence of a cognate ligand, RXR self-associates into tetramers, thereby silencing its own transcriptional activity. It was also shown that the tetramerization region of RXR critically contains two of three consecutive phenylalanine residues found in helix 11 (H11) of the receptor's ligand binding domain. Mutation of these residues abolishes the ability of RXR to form tetramers but also results in a receptor that is defective in its ligand-induced transcriptional activity. These observations suggest that the region may be involved in the association of RXR with transcriptional coactivators. Here, it is demonstrated that mutation of the H11 phenylalanine residues diminishes the ability of RXR to associate with the p160 coactivators TIF2 and p/CIP, but has little effect on ligand-dependent interactions of the receptor with the unrelated coactivator TIF1. It is further shown that a peptide comprised of the H11 sequence effectively competes with RXR for binding of TIF2 but not of TIF1. Finally, transactivation assays demonstrate that the defective transcriptional activity of the H11 mutant can be rescued by ectopic expression of TIF1 but not of TIF2. Taken together, the results indicate that H11 is directly involved in stabilizing the interactions of RXR with p160 coactivators, but is not required for association with TIF1. This region is thus a novel coactivator interaction surface which selectively mediates the association of RXR with transcriptional coactivators. PMID- 11851397 TI - Inositol pyrophosphates are required for DNA hyperrecombination in protein kinase c1 mutant yeast. AB - Diphosphoinositol pentakisphosphate (InsP(7)) and bis-diphosphoinositol tetrakisphosphate (InsP(8)) contain energetic pyrophosphate groups, occur throughout animal and plant kingdoms, and are synthesized by a recently cloned family of inositol hexakisphosphate kinases (InsP(6)Ks). We report that these inositol pyrophosphates mediate homologous DNA recombination in yeast S. cerevisae. Hyperrecombination, caused by altered protein kinase C1 (PKC1), is lost in yeast with deletion of yeast InsP(6)K (yInsP(6)K) and can be restored selectively by catalytically active yeast or mammalian InsP(6)Ks. Inositol pyrophosphates are required for two forms of hyperrecombination that differ in mechanism, suggesting some generalities for actions of inositol pyrophosphates in recombination. PMID- 11851398 TI - Identification of an active site ligand for a group I ribozyme catalytic metal ion. AB - The transition state of the group I intron self-splicing reaction is stabilized by three metal ions. The functional groups within the intron substrates (guanosine and an oligoribonucleotide mimic of the 5'-exon) that coordinate these metal ions have been systematically defined through a series of metal ion specificity switch experiments. In contrast, the catalytic metal ligands within the ribozyme active site are unknown. In an effort to identify them, stereospecific (R(P) or S(P)) single-site phosphorothioate substitutions were introduced at five phosphates predicted to be in the vicinity of the catalytic center (A207, C208, A304, U305, and A306) within the Tetrahymena intron. Of the 10 ribozymes that were studied, four phosphorothioate substitutions (A207 S(P), C208 S(P), A306 R(P), and A306 S(P)) exhibited a significant reduction in the cleavage rate. Only the effect of the C208 S(P) phosphorothioate substitution could be significantly rescued by the addition of a thiophilic metal ion, either Mn(2+) or Zn(2+), when tested with an all-oxy substrate. The effect was not rescued with Cd(2+). To determine if one of the catalytic metal ions is coordinated to the C208 pro-S(P) oxygen, the phosphorothioate-substituted ribozymes were also assayed using oligonucleotide substrates with a 3' phosphorothiolate or an S(P) phosphorothioate substitution at the scissile phosphate. This resulted in a second metal specificity switch, in that Mn(2+) or Zn(2+) no longer rescued the C208 S(P) ribozyme, but Cd(2+) provided efficient rescue in the context of either sulfur-containing substrate. The 3'-oxygen and the pro-S(P) oxygen of the scissile phosphate are both known to coordinate the same metal ion, M(A), which stabilizes the negative charge on the leaving group 3'-oxygen in the transition state. Taken together, these data suggest that metal M(A) is coordinated to the C208 pro-S(P) phosphate oxygen, which constitutes the first functional link between a specific catalytic metal ion and a particular functional group within the group I ribozyme active site. PMID- 11851399 TI - Correlation of the kinetics of finger domain mutants in RB69 DNA polymerase with its structure. AB - We have estimated pre-steady-state kinetic parameters for the addition of a single nucleotide residue by a set of RB69 DNA polymerase mutants in which four highly conserved residues in the fingers domain have been replaced by Ala. The relationship between the kinetic constants exhibited by the mutants and the structure of the ternary complex [Franklin, M., Wang, J., and Steitz T. (2001) Cell 105, 657-667] was consistent with the following sets of interactions between the conserved residues and oxygen atoms in the triphosphate portion of the incoming dNTP: (i) the epsilon-amino group of K560 contacts oxygen atoms of the alpha- and gamma-phosphates, (ii) the amide side chain of Asn 564 forms a hydrogen bond via a water molecule with the nonbridging oxygen of the beta phosphate, and (iii) the epsilon-amino and delta-guanidino groups of K486 and R482, respectively, contact the nonbridging oxygens of the gamma-phosphate. We have also determined the pre-steady-state kinetic parameters for the addition of both dCTP and dCDP onto a 13/20mer primer/template with an exo(-) derivative of RB69 DNA polymerase and have shown that the deoxynucleoside diphosphate can be incorporated, in contrast to the behavior of the Klenow fragment which cannot use dCDP as a substrate. We have shown that, with RB69 DNA polymerase, in contrast to the Klenow fragment, there is no inhibition of the primer-extension reaction by incoming NTPs having either noncomplementary bases or ribo- instead of a deoxyribose moieties. This implies that the mode of recognition of incoming dNTPs and triggering of the conformational change, which is thought to occur prior to the chemical step, differs between these two enzymes. PMID- 11851400 TI - Characterization of the residues involved in the human alpha-thrombin-haemadin complex: an exosite II-binding inhibitor. AB - Haemadin is a 57-amino acid thrombin inhibitor from the land-living leech Haemadipsa sylvestris, whose structure has recently been determined in complex with human alpha-thrombin. Here we communicate the effect of ionic strength on the kinetics of the inhibition of human alpha-thrombin by haemadin, by using thrombin mutants modified in exosite II. Data analysis has allowed both the ionic and nonionic binding contributions to be ascertained, with the nonionic component being virtually the same for all of the thrombins that have been examined, while the ionic binding energy contributions varied from molecule to molecule. In the case of the native human alpha-thrombin-haemadin complex, ionic interactions contribute -17 kJ/mol to the Gibbs free energy of binding, this being the equivalent of up to six salt bridges. These salt bridges make up 20% of the total binding energy at zero ionic strength, and this has been attributed to the C terminal tail alone. In addition, the contributions of the N-terminal and C terminal regions of haemadin to its tight binding have been ascertained by using derivatives of both haemadin and thrombin. Limited proteolysis using formic acid produced haemadin cleaved between residues 40 and 41, removing the majority of the C-terminal tail. This truncated haemadin displayed a 20000-fold reduced affinity for thrombin, and was no longer a tight binding inhibitor. A form of thrombin in which the active site serine has been blocked by diisopropyl fluorophosphate binds to haemadin, but with a 72000-fold reduced affinity, indicating that the N-terminus is more important than the C-terminus for strong binding. PMID- 11851401 TI - Analysis of the role of the interleukin-2 receptor gamma chain in ligand binding. AB - Interleukin-2 is the primary T cell growth factor secreted by activated T cells. IL-2 is an alpha-helical cytokine that binds to a multisubunit receptor expressed on the surface of a variety of cell types. IL-2Ralpha, IL-2Rbeta, and IL-2Rgammac receptor subunits expressed on the surface of cells may aggregate to form distinct binding sites of differing affinities. IL-2Rgammac was the last receptor subunit to be identified. It has since been shown to be shared by at least five other cytokine receptors. In this study, we have probed the role of IL-2Rgammac in the assembly of IL-2R complexes and in ligand binding. We demonstrate that in the absence of ligand IL-2Rgammac does not possess detectable affinity for IL 2Ralpha, IL-2Rbeta, or the pseudo-high-affinity binding site composed of preformed IL-2Ralpha/beta. We also demonstrate that IL-2Rgammac possesses an IL-2 dependent affinity for IL-2Rbeta and IL-2Ralpha/beta. We performed a detailed biosensor analysis to examine the interaction of soluble IL-2Rgammac with IL-2 bound IL-2Rbeta and IL-2-bound IL-2Ralpha/beta. The kinetic and equilibrium constants for sIL-2Rgammac binding to these two different liganded complexes were similar, indicating that IL-2Ralpha does not play a role in recruitment of IL 2Rgammac. We also determined that the binding of IL-2 to the isolated IL-2Rgammac was very weak (approximate K(D) = 0.7 mM). The experimental methodologies and principles derived from these studies can be extended to at least five other cytokines that share IL-2Rgammac as a receptor subunit. PMID- 11851402 TI - Crystallographic studies on the role of the C-terminal segment of human angiogenin in defining enzymatic potency. AB - Human angiogenin (Ang) is an RNase in the pancreatic RNase superfamily that induces angiogenesis. Its catalytic activity is comparatively weak, but nonetheless critical for biological activity. The crystal structure of Ang has shown that enzymatic potency is attenuated in part by the obstructive positioning of Gln117 within the B(1) pyrimidine binding pocket, and that the C-terminal segment of residues 117-123 must reorient for Ang to bind and cleave RNA. The native closed conformation appears to be stabilized by Gln117-Thr44 and Asp116 Ser118 hydrogen bonds, as well as hydrophobic packing of Ile119 and Phe120. Consistent with this view, Q117G, D116H, and I119A/F120A variants are 4-30-fold more active than Ang. Here we have determined crystal structures for these variants to examine the structural basis for the activity increases. In all three cases, the C-terminal segment remains obstructive, demonstrating that none of the residues that has been replaced is essential for maintaining the closed conformation. The Q117G structure shows no changes other than the loss of the side chain of residue 117, whereas those of D116H and I119A/F120A reveal C terminal perturbations beyond the replacement site, suggesting that the native closed conformation has been destabilized. Thus, the interactions of Gln117 seem to be less important than those of residues 116, 119, and 120 for stabilization. In D116H, His116 does not replicate either of the hydrogen bonds of Asp116 with Ser118 and instead forms a water-mediated interaction with catalytic residue His114; residues 117-121 deviate significantly from their positions in Ang. In I119A/F120A, the segment of residues 117-123 has become highly mobile and all of the interactions thought to position Gln117 have been weakened or lost; the space occupied by Phe120 in Ang is partially filled by Arg101, which has moved several angstroms. A crystal structure was also determined for the deletion mutant des(121-123), which has 10-fold reduced activity toward large substrates. The structure is consistent with the earlier proposal that residues 121-123 form part of a peripheral substrate binding subsite, but also raises the possibility that changes in the position of another residue, Lys82, might be responsible for the decreased activity of this variant. PMID- 11851403 TI - Crystal structure of the tetrameric cytidine deaminase from Bacillus subtilis at 2.0 A resolution. AB - Cytidine deaminases (CDA, EC 3.5.4.5) are zinc-containing enzymes in the pyrimidine salvage pathway that catalyze the formation of uridine and deoxyuridine from cytidine and deoxycytidine, respectively. Two different classes have been identified in the CDA family, a homodimeric form (D-CDA) with two zinc ions per dimer and a homotetrameric form (T-CDA) with four zinc ions per tetramer. We have determined the first structure of a T-CDA from Bacillus subtilis. The active form of T-CDA is assembled of four identical subunits with one active site apiece. The subunit of D-CDA is composed of two domains each exhibiting the same fold as the T-CDA subunits, but only one of them contains zinc in the active site. The similarity results in a conserved structural core in the two CDA forms. An intriguing difference between the two CDA structures is the zinc coordinating residues found at the N-terminal of two alpha-helices: three cysteine residues in the tetrameric form and two cysteine residues and one histidine residue in the dimeric form. The role of the zinc ion is to activate a water molecule and thereby generate a hydroxide ion. How the zinc ion in T-CDA surrounded with three negatively charged residues can create a similar activity of T-CDA compared to D-CDA has been an enigma. However, the structure of T-CDA reveals that the negative charge caused by the three ligands is partly neutralized by (1) an arginine residue hydrogen-bonded to two of the cysteine residues and (2) the dipoles of two alpha-helices. PMID- 11851404 TI - Positively charged amino acids are essential for electron transfer and protein protein interactions in the soluble methane monooxygenase complex from Methylococcus capsulatus (Bath). AB - The soluble methane monooxygenase (sMMO) complex from Methylococcus capsulatus (Bath) catalyses oxygen- and NAD(P)H-dependent oxygenation of methane, propene, and other substrates. Whole-complex sMMO oxygenase activity requires all three sMMO components: the hydroxylase, the reductase, and protein B. Also, in the presence of hydrogen peroxide, the hydroxylase alone catalyzes substrate oxygenation via the peroxide shunt reaction. We investigated the effect of amine cross-linking on hydroxylase activity to probe the role of a gross conformational change that occurs in the hydroxylase upon binding of the other protein components. The cross-linker inhibited hydroxylase activity in the whole complex, but this effect was due to covalent modification of primary amine groups rather than cross-linking. Covalent modification of arginine side-chains on the hydroxylase had a similar effect, but, most remarkably, neither form of modification affected the activity of the hydroxylase via the peroxide shunt reaction. It was shown that covalent modification of positively charged groups on the hydroxylase, which occurred at multiple sites, interfered with its physical and functional interactions with protein B and with the passage of electrons from the reductase. These results indicate that protein B and the reductase of the sMMO complex interact via positively charged groups on the surface of the hydroxylase to induce a conformational change that is necessary for delivery of electrons into the active site of the hydroxylase. Modification of positively charged groups on protein B had no effect on its function, consistent with the hypothesis that positively charged groups on the hydroxylase interact with negative charges on protein B. Thus, we have discovered a means of specifically inactivating the interactions between the sMMO complex while preserving the catalytic activity of the hydroxylase active site which provides a new method of studying intercomponent interactions within sMMO. PMID- 11851405 TI - Redox-dependent regulation of the conformation and function of human heat shock factor 1. AB - We present here evidence that redox-dependent thiol-disulfide exchange can provide a mechanism regulating the conformation and activity of the human heat shock transcription factor 1 (HSF1). Diamide and dithiothreitol were reagents used respectively to promote and reverse disulfide cross-link, and the resolution and detection of redox conformers of HSF1 were done according to recently published methods [Manalo, D. J., and Liu, A. Y.-C. (2001) J. Biol. Chem. 276, 23554-23561]. We showed that preincubation of the latent HSF1 monomer with diamide inhibited the in vitro heat-induced activation and trimerization of HSF1 and caused the formation of ox-hHSF1, a compact, disulfide cross-linked HSF1 conformer detectable in immuno-Western blot assay. These effects of diamide were dose-dependent and were rapidly and quantitatively reversed by dithiothreitol. Cysteine site-specific mutants of HSF1 were constructed and used to determine which of the five cysteine residues may be engaged in disulfide cross-link. Analysis of the in vitro transcribed and translated HSF1 proteins showed that while mutation of C1 (amino acid 36) and C2 (amino acid 103) had no effect on the redox sensitivity of HSF1, the mutation of C3 (amino acid 153) or double mutation of C4 and C5 (amino acids 373 and 378, respectively) to serine rendered HSF1 insensitive to diamide and prevented its conversion to ox-HSF1. HSF1 with a single cysteine to serine mutation at either the C4 or C5 position gave different ox-HSF1 conformers in the presence of diamide, suggesting that C3 could be disulfide cross-linked to either C4 or C5. The possibility that HSF1 may have integrated redox chemistry of cysteine sulfhydryl into its functional response in higher mammalian cells is discussed. PMID- 11851406 TI - Macromolecular import into Escherichia coli: the TolA C-terminal domain changes conformation when interacting with the colicin A toxin. AB - Various macromolecules such as bacteriotoxins and phage DNA parasitize some envelope proteins of Escherichia coli to infect the bacteria. A two-step import mechanism involves the primary interaction with an outer membrane receptor or with a pilus followed by the translocation across the outer membrane. However, this second step is poorly understood. It was shown that the TolA, TolQ, and TolR proteins play a critical role in the translocation of group A colicins and filamentous bacteriophage minor coat proteins (g3p). Translocation of these proteins requires the interaction of their N-terminal domain with the C-terminal domain of TolA (TolAIII). In this work, short soluble TolAIII domains were overproduced in the cytoplasm and in the periplasm of E. coli. In TolAIII, the two cysteine residues were found to be reduced in the cytoplasmic form and oxidized in the periplasmic form. The interaction of TolAIII with the N-terminal domain of colicin A (ATh) is observed in the presence and in the absence of the disulfide bridge. The complex formation of TolAIII and ATh was found to be independent of the ionic strength. An NMR study of TolAIII, both free and bound, shows a significant structural change when interacting with ATh, in the presence or absence of the disulfide bridge. In contrast, such a structural modification was not observed when TolAIII interacts with g3p N1. These results suggest that bacteriotoxins and Ff bacteriophages parasitize E. coli using different interactions between TolA and the translocation domain of the colicin and g3p protein, respectively. PMID- 11851407 TI - Dissection of the pathway of molecular recognition by calmodulin. AB - Amide hydrogen exchange has been used to examine the structural dynamics and energetics of the interaction of a peptide corresponding to the calmodulin binding domain of smooth muscle myosin light chain kinase (smMLCKp) with calcium saturated calmodulin. Heteronuclear NMR (15)N-(1)H correlation spectroscopy was used to quantify amide proton exchange rates of the uniformly (15)N-labeled domain bound to calmodulin. A key feature of a proposed model for molecular recognition by calmodulin [Ehrhardt et al. (1995) Biochemistry 34, 2731-2738] is tested by examination of the dependence of amide hydrogen exchange on applied hydrostatic pressure. Hydrogen exchange rates and corresponding protection factors (1/K(op)) for individual amide protons of the bound smMLCKp domain span 5 orders of magnitude at ambient pressure. Individual protection factors decrease significantly in a linear fashion with increasing hydrostatic pressure. A common pressure dependence is revealed by a constant large negative volume change across the residues comprising the core of the bound helical domain. The pattern of protection factors and their response to hydrostatic pressure is consistent with a structural reorganization that results in the concerted disruption of ion pairs between calmodulin and the bound domain. These observations reinforce a model for the molecular recognition pathway where formation of the initial encounter complex is followed by helix-coil transitions in the bound state and subsequent concerted formation of the extensive ion pair network defining the intermolecular contact surface between CaM and the target domain in the final, compact complex structure. PMID- 11851408 TI - A novel restricted photoaffinity spin-labeled non-nucleoside ATP analogue as a covalently attached reporter group of the active site of Myosin subfragment 1. AB - The photoaffinity spin-labeled non-nucleoside ATP analogue, 2-(4-azido-2 nitrophenyl)amino-2,2-(1-oxyl-2,2,6,6-tetramethyl-4 piperidylidene)di(oxymethylene)ethyl triphosphate (SSL-NANTP), has been shown to be a substrate for skeletal mysoin subfragment 1 (S1) that can be photoincorporated at the active site of S1 [Chen, X., et al. (2000) Bioconjugate Chem. 11, 725-733]. Electron paramagnetic resonance spectroscopy shows that the probe undergoes restricted motion with respect to the protein. The parent compound, NANTP (2-[(4-azido-2-nitrophenyl)amino]ethyl triphosphate), is specifically photoincorporated at Trp-130 on the amino-terminal 23 kDa tryptic fragment in rabbit skeletal myosin. Surprisingly, amino acid sequence analysis shows that SSL-NANTP is photoincorporated on the carboxy-terminal 20 kDa tryptic fragment at Lys-681 on the side opposite Trp-130 in the nucleotide pocket. This is the first direct evidence showing that this residue in the 20 kDa tryptic fragment is close enough to the active site to be photolabeled by trapped ATP analogues. After actin treatment in the presence of MgATP, SSL-NANDP-labeled myosin S1 had normal ATPase activity, indicating that photolabeling did not significantly alter the enzymatic properties of S1. Photoincorporated SSL-NANDP was bound inside the nucleotide site of S1, with an effective concentration of 20 mM as judged by the concentration of MgADP needed to displace it. Molecular dynamics simulations suggest that the ability of NANTP and SSL-NANTP to photolabel different sites results from different orientations of the phenyl ring in the active site. For SSL-NANTP, the p-azido group on the phenyl ring points toward Lys-681. For NANTP, it points in the opposite direction toward Trp-130. PMID- 11851409 TI - Structural mechanism of enoyl-CoA hydratase: three atoms from a single water are added in either an E1cb stepwise or concerted fashion. AB - We have determined the crystal structure of the enzyme enoyl-CoA hydratase (ECH) from rat liver with the bound substrate 4-(N,N-dimethylamino)cinnamoyl-CoA using X-ray diffraction data to a resolution of 2.3 A. In addition to the thiolester substrate, the catalytic water, which is added in the hydration reaction, has been modeled into well-defined electron density in each of the six active sites of the physiological hexamer within the crystallographic asymmetric unit. The catalytic water bridges Glu(144) and Glu(164) of the enzyme and has a lone pair of electrons poised to react with C(3) of the enzyme-bound alpha,beta-unsaturated thiolester. The water molecule, which bridges two glutamate residues, is reminiscent of the enolase active site. However, unlike enolase, which has a lysine available to donate a proton, there are no other sources of protons available from other active site residues in ECH. Furthermore, an analysis of the hydrogen-bonding network of the active site suggests that both Glu(144) and Glu(164) are ionized and carry a negative charge with no reasonable place to have a protonated carboxylate. This lack of hydrogen-bonding acceptors that could accommodate a source of a proton, other than from the water molecule, leads to a hypothesis that the three atoms from a single water molecule are added across the double bond to form the hydrated product. The structural results are discussed in connection with details of the mechanism, which have been elucidated from kinetics, site-directed mutagenesis, and spectroscopy of enzyme-substrate species, in presenting an atomic-resolution mechanism of the reaction. Contrary to the previous interpretation, the structure of the E-S complex together with previously determined kinetic isotope effects is consistent with either a concerted mechanism or an E1cb stepwise mechanism. PMID- 11851410 TI - Polarization of cinnamoyl-CoA substrates bound to enoyl-CoA hydratase: correlation of (13)C NMR with quantum mechanical calculations and calculation of electronic strain energy. AB - When alpha,beta-unsaturated substrates bind to the active site of enoyl-CoA hydratase, large spectral changes can be observed [D'Ordine, R. L., et al. (1994) Biochemistry 33, 12635-12643]. The differences in the isotropic magnetic shieldings of the free and active site-bound forms of the carbonyl, alpha-, and beta-carbons of the substrates, hexadienoyl-CoA, cinnamoyl-CoA, and (N,N-dimethyl p-amino)cinnamoyl-CoA have been experimentally determined. The carbonyl and beta carbons are all deshielded, while the alpha-carbons show increased shielding. These chemical shift perturbations are interpreted to suggest that the pi electrons of the enoyl thiolester are polarized when bound at the active site. Using the crystal structure of (N,N-dimethyl-p-amino)cinnamoyl-CoA bound at the enzyme active site, the shielding tensors were calculated at three different levels of theory, up to a density functional theory model that included all of the contiguous active site residues. These calculations successfully reproduced the observed spectral changes and permitted the electronic polarization of the substrate to be quantified as an electron density difference map. The calculated electron density difference confirms the loss of electrons at the electrophilic beta-carbon and carbonyl carbon, while a slight increase in electron density at the alpha-carbon where proton donation occurs during the hydration reaction and a larger increase in electron density at the carbonyl oxygen are predicted. The energy required to polarize the electrons to the observed extent was calculated to be 3.2 kcal/mol. The force that provides the requisite energy for the polarization is the interaction of the electric field generated by the protein at the enzyme active site with the polarizable electrons of the substrate. Because the induced electronic polarization is along the predicted reaction pathway, the extent of substrate activation by the induced electronic strain is catalytically relevant. PMID- 11851411 TI - Characterization of the unfolding of ribonuclease a by a pulsed hydrogen exchange study: evidence for competing pathways for unfolding. AB - The unfolding of ribonuclease A was studied in 5.2 M guanidine hydrochloride at pH 8 and 10 degrees C using multiple optical probes, native-state hydrogen exchange (HX), and pulse labeling by hydrogen exchange. First, native-state HX studies were used to demonstrate that the protein exists in two slowly interconverting forms under equilibrium native conditions: a predominant exchange incompetent N form and an alternative ensemble of conformations, N(I), in which some amide hydrogens are fully exposed to exchange. Pulsed HX studies indicated that, during unfolding, the rates of exposure to exchange with solvent protons were similar for all backbone NH probe protons. It is shown that two parallel routes of unfolding are available to the predominant N conformation as soon as it encounters strong unfolding conditions. A fraction of molecules appears to rapidly form N(I) on one route. On the other route an exchange-incompetent intermediate state ensemble, I(U)(2), is formed. The kinetics of unfolding measured by far-UV circular dichroism (CD) were faster than those measured by near-UV CD and intrinsic tyrosine fluorescence of the protein. The logarithms of the rate constants of the unfolding reaction measured by all three optical probes also showed a nonlinear dependence on GdnHCl concentration. All of the data suggest that N(I) and I(U)(2) are nativelike in their secondary and tertiary structures. While N(I) unfolds directly to the fully exchange-competent unfolded state (U), I(U)(2) forms another intermediate I(U)(3) which then unfolds to U. I(U)(3) is devoid of all native alpha-helical secondary structure and has only 30% of the tertiary interactions still intact. Since the rates of global unfolding measured by near-UV CD and fluorescence agree well with the rates of exposure determined for all of the backbone NH probe protons, it appears that the rate-limiting step for the unfolding of RNase A is the dissolution of the entire native tertiary structure and penetration of water into the hydrophobic core. PMID- 11851412 TI - Backbone dynamics of the ribonuclease binase active site area using multinuclear ((15)N and (13)CO) NMR relaxation and computational molecular dynamics. AB - The nano-pico second backbone dynamics of the ribonuclease binase, homologous to barnase, is investigated with (15)N, (13)C NMR relaxation at 11.74 and 18.78 T and with a 1.1 ns molecular dynamics simulation. The data are compared with the temperature factors reported for the X-ray structure of this enzyme. The molecular dynamics and X-ray data correspond well and predict motions in the loops 56-61 and 99-104 that contain residues that specifically recognize substrate and are catalytic (His101), respectively. In contrast, the (15)N relaxation data indicate that these loops are mostly ordered at the nano-pico second time scale. Nano-pico second motions in the recognition loop 56-61 are evident from (13)CO-(13)C cross relaxation data, but the mobility of the catalytic loop 99-104 is not detected by (13)CO cross relaxation either. From the results of this and previous work [Wang, L., Pang, Y., Holder, T., Brender, J. R., Kurochkin, A., and Zuiderweg, E. R. P. (2001) Proc. Natl. Acad. Sci. U.S.A., 98, 7684-7689], the following dynamical characterization of the active site area of binase emerges: a beta sheet, rigid at all probed time scales, supports the catalytic residue Glu 72. Both substrate-encapsulating loops are mobile on both fast and slow time scales, but the fast motions of the loop which contains the other catalytic residue, His 101, as predicted by B-factors and computational molecular dynamics is not detected by NMR relaxation. This work strongly argues for the use of several measures in the study of protein dynamics. PMID- 11851413 TI - Nuclear wavepacket motion between P and P(+)B(A)(-) potential surfaces with subsequent electron transfer to H(A) in bacterial reaction centers. 1. Room temperature. AB - Formation and coherent propagation of nuclear wavepackets on potential energy surfaces of the excited state of the primary electron donor P and of the charge transfer states P(+)B(A)(-) and P(+)H(A)(-) were studied in native and pheophytin modified Rhodobacter sphaeroides R-26 reaction centers (RCs) induced by 25 fs excitation (where B(A) and H(A) are the primary and secondary electron acceptors, respectively). The processes were monitored by measuring coherent oscillations in kinetics of the time evolution of the stimulated emission band of P at 935 nm, of the absorption band of B(A)(-) at 1020 nm, and of the bleaching band of H(A) at 760 nm. It was found that the nuclear wavepacket motion on the 130-140 cm(-1) surface of P is directly induced by light absorption in P. When the wavepacket approaches the intersection between P and P(+)B(A)(-) surfaces at 120 and 380 fs delays, the formation of intermediate mixed-state emitting light at 935 nm (P) and absorbing light at 1020 nm (P(+)B(A)(-)) takes place. At the latter time, the wavepacket is transferred to the 32 cm(-1) mode which can belong to the P hypersurface effectively transferring the wavepacket to the P(+)B(A)(-) surface or can represent a diabatic surface which is formed by the states P and P(+)B(A)( ). The wavepacket motion on the P(+)B(A)(-) surface or on the P(+)B(A)(-) part of the mixing surface is accompanied by irreversible electron transfer to H(A). This process is monitored by the kinetics of 1020 nm band development and 760 nm band bleaching (delayed with respect to 1020 nm band development) which both have the enhanced 32 cm(-1) mode in Fourier transform (FT) spectra. The mechanism of wavepacket transfer from the 130-140 cm(-1) to the 32 cm(-1) mode is discussed. PMID- 11851414 TI - Time-resolved step-scan Fourier transform infrared spectroscopy of the CO adducts of bovine cytochrome c oxidase and of cytochrome bo(3) from Escherichia coli. AB - We have used cryogenic difference FTIR and time-resolved step-scan Fourier transform infrared (TR-FTIR) spectroscopies to explore the redox-linked proton pumping mechanism of heme-copper respiratory oxidases. These techniques are used to probe the structure and dynamics of the heme a(3)-Cu(B) binuclear center and the coupled protein structures in response to the photodissociation of CO from heme Fe and its subsequent binding to and dissociation from Cu(B). Previous cryogenic (80 K) FTIR CO photodissociation difference results were obtained for cytochrome bo(3), the ubiquinol oxidase of Escherichia coli [Puustinen, A., et al. (1997) Biochemistry 36, 13195-13200]. These data revealed a connectivity between Cu(B) and glutamic acid E286, a residue which has been implicated in proton pumping. In the current work, the same phenomenon is observed using the CO adduct of bovine cytochrome aa(3) under cryogenic conditions, showing a perturbation of the equivalent residue (E242) to that in bo(3). Furthermore, using time-resolved (5 micros resolution) step-scan FTIR spectroscopy at room temperature, we observe the same spectroscopic perturbation in both cytochromes aa(3) and bo(3). In addition, we observe evidence for perturbation of a second carboxylic acid side chain, at higher frequency in both enzymes at room temperature. The high-frequency feature does not appear in the cryogenic difference spectra, indicating that the perturbation is an activated process. We postulate that the high-frequency IR feature is due to the perturbation of E62 (E89 in bo(3)), a residue near the opening of the proton K-channel and required for enzyme function. The implications of these results with respect to the proton pumping mechanism are discussed. Finally, a fast loss of over 60% of the Cu(B)-CO signal in bo(3) is observed and ascribed to one or more additional conformations of the enzyme. This fast conformer is proposed to account for the uninhibited reaction with O(2) in flow-flash experiments. PMID- 11851415 TI - Cation-induced stabilization of the engineered cation-binding loop in cytochrome c peroxidase (CcP). AB - We have previously shown that the K(+) site found in the proximal heme pocket of ascorbate peroxidase (APX) could be successfully engineered into the closely homologous cytochrome c peroxidase (CcP) [Bonagura et al., (1996) Biochemistry 35, 6107-6115; Bonagura et al. (1999) Biochemistry 38, 5538-5545]. In addition, specificity could be switched to binding Ca(2+) as found in other peroxidases [Bonagura et al. (1999) J. Biol. Chem. 274, 37827-37833]. The introduction of a proximal cation-binding site also promotes conversion of the Trp191 containing cation-binding loop from a "closed" to an "open" conformer. In the present study we have changed a crucial hinge residue of the cation-binding loop, Asn195, to Pro which stabilizes the loop, albeit, only in the presence of bound K(+). The crystal structure of this mutant, N195PK2, has been refined to 1.9 A. As predicted, introduction of this crucial hinge residue stabilizes the cation binding loop in the presence of the bound K(+). As in earlier work, the characteristic EPR signal of Trp191 cation radical becomes progressively weaker with increasing [K(+)] and the lifetime of the Trp191 radical also has been considerably shortened in this mutant. This mutant CcP exhibits reduced enzyme activity, which could be titrated to lower levels with increasing [K(+)] when horse heart cytochrome c is the substrate. However, with yeast cytochrome c as the substrate, the mutant was as active as wild-type at low ionic strength, but 40-fold lower at high ionic strength. We attribute this difference to a change in the rate-limiting step as a function of ionic strength when yeast cytochrome c is the substrate. PMID- 11851416 TI - Exploring the energy profile of the Q(A)(-) to Q(B) electron transfer reaction in bacterial photosynthetic reaction centers: pH dependence of the conformational gating step. AB - Both large- and small-scale conformational changes are needed as proteins carry out reactions. However, little is known about the identity, energy of, and barriers between functional substates on protein reaction coordinates. In isolated bacterial photosynthetic reaction centers, the electron transfer from the reduced primary quinone, Q(A)(-), to the secondary quinone, Q(B), is rate limited by conformational changes at low pH and by proton binding at high pH. The kinetics and thermodynamics of this reaction were determined between 200 and 300 K from pH 6 to pH 10.5. A model with two substates of the reactant, P(+)Q(A)( )Q(B), one protonated (state A) and one unprotonated (alpha), and one state of the product, P(+)Q(A)Q(B)(-) (B), was able to simulate the dependence of the rate on temperature and pH fairly well. The equilibrium between the three states were measured in situ at each temperature. Proton binding (alpha to A transition) has a favorable DeltaH and unfavorable DeltaS as does the conformational changes required for electron transfer at low pH (A to B). The pK for the A to alpha transition is 9.7 at room temperature, consistent with previous measurements, and equivalent to 13.5 at 200 K. The activation barriers were determined for each transition. Both the alpha to A and the A to B transitions are limited primarily by the activation enthalpy with modest DeltaS. PMID- 11851417 TI - N-terminus of the photosystem II manganese stabilizing protein: effects of sequence elongation and truncation. AB - The importance of the N-terminal domain of manganese stabilizing protein in binding to photosystem II has been previously demonstrated [Eaton-Rye and Murata (1989) Biochim. Biophys. Acta 977, 219-226; Odom and Bricker (1992) Biochemistry 31, 5616-5620]. In this paper, we report results from a systematic study of functional and structural consequences of N-terminal elongation and truncation of manganese stabilizing protein. Precursor manganese stabilizing protein is the unprocessed wild-type protein, which carries an N-terminal extension of 84 amino acids in the form of its chloroplastic signal peptide. Despite its increased size, this protein is able to reconstitute O(2) evolution activity to levels observed with the mature, processed protein, but it also binds nonspecifically to PSII. Truncation of wild-type manganese stabilizing protein by site-directed mutagenesis to remove three N-terminal amino acids, resulting in a mutant called DeltaG3M, causes no loss of activity reconstitution, but this protein also exhibits nonspecific binding. Further truncation of the wild-type protein by ten N-terminal amino acids, producing DeltaE10M, limits binding of manganese stabilizing protein to 1 mol/mol of photosystem II and decreases activity reconstitution to about 65% of that obtained with the wild-type protein. Because two copies of wild type normally bind to photosystem II, amino acids in the domain (4)K-(10)E must be involved in the binding of one copy of manganese stabilizing protein to photosystem II. Spectroscopic analysis (CD and UV spectra) reveals that N-terminal elongation and deletion of manganese stabilizing protein influence its overall conformation, even though secondary structure content is not perturbed. Our data suggest that the solution structure of manganese stabilizing protein attains a more compact solution structure upon removal of N terminal amino acids. PMID- 11851418 TI - Cytochrome P450 3A4-mediated oxidative conversion of a cyano to an amide group in the metabolism of pinacidil. AB - The conversion of nitriles to amides is generally considered to be a hydrolytic process that does not involve redox chemistry. We demonstrate here that cytochrome P450 (CYP) is responsible for the conversion of the cyano group of pinacidil to the corresponding amide. The reaction in human liver microsomes was NADPH-dependent and was nearly completely inhibited by an anti-CYP3A4 antibody. Incubations of pinacidil with recombinant CYP enzymes confirm that CYP3A4 is the principal catalyst of this reaction. The kinetics of pinacidil amide formation by CYP3A4 yielded an apparent K(m) of 452 +/- 33 microM and k(cat) of 0.108 min(-1) (k(cat)/K(m) = 0.238 mM(-1).min(-1)). Incubation of pinacidil with CYP3A4 in the presence of (18)O(2) or H(2)(18)O showed that the amide carbonyl oxygen derived exclusively from molecular oxygen. The CYP3A4-mediated reaction also was supported by hydrogen peroxide when incubations were carried out in the absence of cytochrome P450 reductase and NADPH. The reaction can be explained by a nucleophilic attack of a deprotonated ferric peroxide intermediate (Fe(3+)-O-O( )) on the carbon atom of the -C triple bond N triple bond to form an Enz-Fe(III) O-O-C(=NH)R intermediate, followed by cleavage of the O-O bond to give pinacidil amide. This nucleophilic addition of an Fe(3+)-O-O(-) intermediate to a -C=N pi bond in a P450 system resembles the analogous reaction catalyzed by the nitric oxide synthases. PMID- 11851419 TI - Stereochemistry of catalysis by the ketoreductase activity in the first extension module of the erythromycin polyketide synthase. AB - Multiple ketoreductase activities play a crucial role in establishing the stereochemistry of the products of modular polyketide synthases (PKSs), but there has been little systematic scrutiny of catalysis by individual ketoreductases. To allow this, a diketide synthase, consisting of the loading module, first extension module, and the chain-terminating thioesterase of the erythromycin producing PKS of Saccharopolyspora erythraea, has been expressed and purified. The DNA encoding the ketoreductase-1 domain in this construct is flanked by unique restriction sites so that another ketoreductase domain can be readily substituted. The purified recombinant diketide synthase catalyzes, at a very low rate (k(cat) equals 2.5 x 10(-3) s(-1)), the specific production of the diketide (2S,3R)-2-methyl-3-hydroxypentanoic acid. The activity of the ketoreductase domain in this model synthase was analyzed using as a model substrate (+/-)-2 methyl-3-oxopentanoic acid N-acetylcysteaminyl (NAC) ester for which k(cat)/K(m) was 21.7 M(-1) s(-1). The NAC thioester of (2S,3R)-2-methyl-3-hydroxypentanoic acid was the major product and was strongly preferred over other stereoisomers as a substrate in the reverse reaction. The bicyclic ketone (9RS)-trans-1-decalone, a known substrate for ketoreductase in fatty acid synthase, was found also to be an effective substrate for the ketoreductase of the diketide synthase. Only the (9R)-trans-1-decalone was reduced, selectively and reversibly, to the (1S,9R) trans-decalol. The stereochemical course of reduction and oxidation is exactly as found previously for the ketoreductase of animal fatty acid synthase, an additional indication of the close similarity of these enzymes. PMID- 11851420 TI - Characterization of the 2-[(R)-2-hydroxypropylthio]ethanesulfonate dehydrogenase from Xanthobacter strain Py2: product inhibition, pH dependence of kinetic parameters, site-directed mutagenesis, rapid equilibrium inhibition, and chemical modification. AB - Although the short-chain dehydrogenase/reductase (SDR) superfamily contains a very large number of members defined in annotated databases and by biochemical and structural studies, very few SDR enzymes have been identified that have a homologous partner catalyzing the same reaction but with an opposite stereospecificity. In the present study we have cloned and expressed one of these enzymes, the 2-[(R)-2-hydroxypropylthio]ethanesulfonate (R-HPC) dehydrogenase, that is part of the coenzyme M-dependent pathway of alkene and epoxide metabolism in Xanthobacter strain Py2. Investigation of the kinetic mechanism using product inhibition suggested that a compulsory-ordered ternary complex mechanism was followed. The pH dependence of k(cat)/K(m) indicated the presence of a single ionizable residue of catalytic importance (pK(a) = 6.9) that was proposed to be Y155 of the catalytic triad. Amino acid substitutions of the putative catalytic triad residues produced inactive enzymes (S142C, Y155F, Y155E, and K159A) or enzyme with a greatly decreased activity (S142A). Inhibitors were investigated as probes of the molecular features of R-HPC that contribute to substrate binding. 2 [(S)-2-Hydroxypropylthio]ethanesulfonate (S-HPC) and 2-(2-methyl-2 hydroxypropylthio)ethanesulfonate were found to be competitive inhibitors of R HPC with K(ic) values close to the K(m) for R-HPC. The arginine-specific modifiers 2,3-butanedione and phenylglyoxal were found to be inactivators, and inactivation could be protected against by the addition of R-HPC. 2,3-Butanedione was found to reduce enzyme activity with R-HPC as a substrate much more dramatically than with substrates that lacked a sulfonate moiety [e.g., 2 propanol, (R)-2-pentanol, and (R)-2-heptanol]. Amino acid analyses of enzyme modified by 2,3-butanedione in the presence and absence of S-HPC suggested protection of a single arginine residue. On the basis of these results, we propose that one or more active site arginines play a key role in substrate binding via an ionic interaction with the sulfonate moiety of R-HPC. PMID- 11851421 TI - Substrate binding determinants of Trypanosoma brucei gamma-glutamylcysteine synthetase. AB - gamma-Glutamylcysteine synthetase (gamma-GCS) catalyzes the ATP-dependent ligation of L-Glu and L-Cys, which is the first step in de novo biosynthesis of the tripeptide glutathione. Recently it was demonstrated that gamma-GCS is a structural homologue of glutamine synthetase (GS), providing the basis to build a model for the gamma-GCS active site [Abbott et al. (2001) J. Biol. Chem. 276, 42099-42107]. Substrate binding determinants in the active site of gamma-GCS have been identified and characterized in the enzyme from the parasitic protozoa Trypanosoma brucei using this model as a guide for site-directed mutagenesis. R366 and R491 were identified as key determinants of L-Glu binding. Mutation of R366 to Ala increases the K(d) for L-Glu by 160-fold and eliminates the positive cooperativity observed for the binding of L-Glu and ATP to the wild-type enzyme, based on a rapid equilibrium random mechanism of substrate binding. Unlike the wild-type enzyme, the R366A mutant enzyme was able to form product using the substrate analogue gamma-aminobutyric acid, suggesting that R366 interacts with the alpha-carboxylate of L-Glu. Mutation of R491 to Ala decreased k(cat) for ATP hydrolysis by 70-fold; however, dipeptide product was only formed in 5% of these turnovers. These data suggest that R491 stabilizes the phosphorylated gamma carboxylate of L-Glu during nucleophilic attack by the L-Cys to form the dipeptide product. T323, R474, and R487 were predicted to be ATP binding determinants. Mutation of each of these residues to Ala increased the apparent K(m) for ATP by 20-100-fold while having only modest effects on k(cat) or the apparent K(m)'s for the other substrates. Finally, mutation of R179, a conserved residue that is present in gamma-GCS, but not in GS, increased the apparent K(m) for both L-Cys and L-Glu. PMID- 11851422 TI - Identification of the general acid/base catalyst of a family 3 beta-glucosidase from Flavobacterium meningosepticum. AB - beta-Glucosidase from Flavobacterium meningosepticum (Fbgl) (also known as Chryseobacterium meningosepticum) has been classified as a member of the family 3 glycohydrolases. It is a retaining enzyme involving a two-step, double displacement mechanism. D247 was shown to function as the nucleophile of the enzymatic reaction [Li, Y.-K., Chir, J., and Chen, F.-Y. (2001) Biochem. J. 355, 835-840]. However, the general acid/base catalyst of this enzyme and of all other family 3 glycohydrolases has not yet been identified. On the basis of amino acid sequence alignment of 15 family 3 enzymes, 11 residues (D71, R129, E132, E136, D137, K168, H169, E177, D247, D458, and E473) are highly conserved. All of these residues are studied by site-directed mutagenesis and kinetic investigation. Analyzing the catalytic power of all mutants reveals E473 residue is the best candidate of the acid/base catalyst. Detailed studies supporting this suggestion are summarized as follows. (1) The k(cat) and K(m) values for the hydrolysis of 2,4-dinitrophenyl beta-D-glucopyranoside (2,4-DNPG) by E473G are reduced 3300- and 900-fold, respectively, compared with those of the wild type (WT). (2) The k(cat) values of E473G-catalyzed hydrolysis are virtually invariant with pH over the range of 5.0-9.0. (3) The activity of E473G with 2,4-DNPG is enhanced by the addition of azide, and beta-glucosyl azide is formed. (4) The k(cat) of the reaction of 2-carboxyphenyl beta-glucoside catalyzed by E473G is comparable to that for hydrolysis by wild-type Fbgl and is 100- and 320-fold better than the k(cat) values for the E473G-catalyzed hydrolysis of 4-carboxyphenyl beta glucoside and the corresponding methyl ester, respectively. (5) The accumulated glucosyl-enzyme intermediate was directly observed by mass analysis in the reaction of 2,4-DNPG with E473G. All of these results confirm that E473 is the general acid/base catalyst of Fbgl. PMID- 11851423 TI - HMGA1 enhances the transcriptional activity and binding of the estrogen receptor to its responsive element. AB - The estrogen receptor (ER) plays a critical role in the development of hormone dependent cancer. Since HMGA1, a member of the "high mobility group" proteins, is overexpressed in certain malignant cells, we investigated the interaction between these nuclear proteins. Transfection of the HMGA1 expression vector increased 2 fold the transcriptional activation of ERE containing promoter by E(2). Furthermore, the HMGA1 protein stimulated severalfold the binding of purified ER to the consensus ERE oligonucleotides in gel mobility shift assays and saturation assays. However, HMGA1 could not bind alone either to consensus or to modified EREs, and the minor groove binding drug distamycin A failed to prevent the synergism between ER and HMGA1. This could suggest that the binding of HMGA1 to DNA was not required for its stimulatory effect. Antibody supershift assays showed that HMGA1 was required for increased binding and suggest a protein protein interaction between those factors. This was confirmed by pull down assay. These data show that HMGA1 acts in concert with the ER to regulate the expression of estrogen responsive genes through a mechanism that does not require direct binding to DNA. These observations may be relevant in malignant cells expressing both proteins. PMID- 11851424 TI - Immobilized small deoxyribozyme to distinguish RNA secondary structures. AB - The RNA folding variation due to one or more mutations leads to different RNA splicing, RNA processing, and translational controls as a result of differences in the primary and higher-ordered structures that interact with other cellular molecules. Thus, distinguishing RNA folding is one of the guides to detect the gene functions related to disease and drug responses. We found, previously, a small Ca(2+)-dependent deoxyribozyme with its site-specific RNA cleavage [Sugimoto, N., Okumoto, Y., and Ohmichi, T. (1999) J. Chem. Soc., Perkin Trans. 2, 1382-1388]. In this study, we report the potential of this deoxyribozyme as a useful tool to distinguish RNA foldings. It is found that the immobilized deoxyribozyme using avidin-biotin interaction cleaves the target site within only single-stranded RNAs. The systematic design for the target RNA hairpin loops shows that the immobilized deoxyribozyme is able to cleave them with a > or =17 nucleotide loop size at only one site under single-turnover conditions. Furthermore, an RNA cleavage reaction is detected using the immobilized deoxyribozyme on a surface plasmon resonance (SPR) sensor chip. These results show that the immobilized deoxyribozymes on a column and on an SPR sensor chip become a novel and useful tool to distinguish the RNA foldings. PMID- 11851425 TI - Transcription activation by ultrabithorax Ib protein requires a predicted alpha helical region. AB - Characterization of their transcription activation domains is critical to understanding functional specificity within the Hox family of proteins. However, few Hox activation domains have been identified and none characterized in detail. In this study, promotor-reporter assays in yeast and Drosophila S2 cell culture were used to refine the boundaries of the activation domain of the Drosophila Hox protein Ultrabithorax (Ubx) and to identify critical elements within this domain. We found that residues 159-242 were sufficient for 50% function, and full transactivation capacity was achieved with inclusion of additional N-terminal sequences. Activation domain sequence and placement relative to the homeodomain differ between Ubx and other Hox proteins, consistent with the possibility that diverse activation mechanisms contribute to functional distinctions in vivo. The essential residues 159-242 in the UbxIb activation domain are predicted to contain a beta-sheet segment followed by an alpha-helix. This putative alpha helical region was established to be necessary, but not sufficient, for transcriptional activation. Disruption of the helix by proline substitutions abolished activation function, while alteration of side chains presented on the surface of this putative helix with alanine or lysine mutations had no significant effect on activity. Collectively, these data indicate that this secondary structural element is a key component in forming an effective activation domain in the UbxIb protein. Interestingly, the alpha-helix critical for transcriptional activation is found only for Ubx orthologs from flies and not other species. The mutant Ubx proteins generated in this study have potential applications in deciphering Hox functions in vivo. PMID- 11851426 TI - Peptide models of folding initiation sites of bovine beta-lactoglobulin: identification of nativelike hydrophobic interactions involving G and H strands. AB - In an attempt to characterize the early folding events in bovine beta lactoglobulin (BLG), a set of peptides, covering the flexible N-terminal region and the stable C-terminus beta-core, was synthesized and analyzed by circular dichroism and by nuclear magnetic resonance in water, trifluoroethanol (TFE), and sodium dodecyl sulfate (SDS) below and above the critical micellar concentration. The role of local and long-range hydrophobic interactions in guiding the folding has been investigated. For the peptide fragment covering the more flexible N terminal region of BLG (beta-strands A, B), where both theoretical predictions and kinetic refolding experiments suggested the formation of non-native alpha helix, no native long-range contacts were identified, and a helical secondary structure was stabilized only in the presence of 25 mM SDS. At variance, in 50% (v/v) TFE, native, long-range hydrophobic interactions were observed in the peptide covering the core region comprising G and H beta-strands. The side chains involved in these interactions form a nativelike hydrophobic cluster, thus suggesting that the GH region may act as the folding initiation site for BLG. This result is reinforced by the identification, in the urea denaturated BLG, of residual structure located at the level of the GH interface, as evidenced by NMR analysis. These results, in excellent agreement with kinetic, thermodynamic, and cold denaturation folding data, once more underline the utmost importance of the GH region for the stability and folding of BLG. Severe aggregation effects prevented the structural analysis of the peptide covering the EFGH region, indicating that this larger segment does not represent an independent folding domain and that the terminal alpha-helix is necessary for stabilizing the BLG folding core. PMID- 11851427 TI - Concerted and stepwise dehydration mechanisms observed in wild-type and mutated Escherichia coli dTDP-glucose 4,6-dehydratase. AB - The conversion of dTDP-glucose into dTDP-4-keto-6-deoxyglucose by Escherichia coli dTDP-glucose 4,6-dehydratase (4,6-dehydratase) takes place in the active site in three steps: dehydrogenation to dTDP-4-ketoglucose, dehydration to dTDP-4 ketoglucose-5,6-ene, and rereduction of C6 to the methyl group. The 4,6 dehydratase makes use of tightly bound NAD(+) as the coenzyme for transiently oxidizing the substrate, activating it for the dehydration step. Dehydration may occur by either of two mechanisms, enolization of the dTDP-4-ketoglucose intermediate, followed by elimination [as proposed for beta-eliminations by Gerlt, J. A., and Gassman, P. G. (1992) J. Am. Chem. Soc. 114, 5928-5934], or a concerted 5,6-elimination of water from the intermediate. To assign one of these two mechanisms, a simultaneous kinetic characterization of glucosyl C5((1)H/(2)H) solvent hydrogen and C6((16)OH/(18)OH) solvent oxygen exchange was performed using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The reaction of the wild-type enzyme is shown to proceed through a concerted dehydration mechanism. Interestingly, mutation of Asp135, the acid catalyst, to Asn or Ala alters the mechanism, allowing enolization to occur to varying extents. While aspartic acid 135 is the acid catalyst for dehydration in the wild-type enzyme, the differential enolization capabilities of D135N and D135A dehydratases suggest an additional role for this residue. We postulate that the switch from a concerted to stepwise dehydration mechanism observed in the aspartic acid variants is due to the loss of control over the glucosyl C5-C6 bond rotation in the active site. PMID- 11851428 TI - Chemically mediated site-specific proteolysis. Alteration of protein-protein interaction. AB - The design and synthesis of a novel iodine-labile serine protease inhibitor was realized by the use of an ecotin analogue containing allylglycine at position 84 in lieu of methionine. Allylglycine-containing ecotins were synthesized by in vitro translation of the ecotin gene containing an engineered nonsense codon (TAG) at the positions of interest. A misacylated suppressor tRNA activated with the unnatural amino acid allylglycine was employed for the suppression of the nonsense codons in a cell-free protein biosynthesizing system, permitting the elaboration of ecotin analogues containing allyglycine at the desired sites. The derived ecotin analogues were capable of inhibiting bovine trypsin with inhibitory constants (K(i)s) comparable to that of wild-type ecotin. Iodine treatment of ecotin analogue Met84(A)Gly resulted in the deactivation of ecotin, caused by peptide backbone cleavage at its P1 reactive site. Upon iodine treatment, active trypsin could be released from the protein complex with ecotin analogue Met84(A)Gly. This constitutes a novel strategy for modulation of serine protease activity and more generally for alteration of protein-protein interaction by a simple chemical reagent. PMID- 11851429 TI - Domain formation in phosphatidylcholine bilayers containing transmembrane peptides: specific effects of flanking residues. AB - Lateral segregation in biological membranes leads to the formation of domains. We have studied the lateral segregation in gel-state model membranes consisting of supported dipalmitoylphosphatidylcholine (DPPC) bilayers with various model peptides, using atomic force microscopy (AFM). The model peptides are derivatives of the Ac-GWWL(AL)(n)WWA-Etn peptides (the so-called WALP peptides) and have instead of tryptophans, other flanking residues. In a previous study, we found that WALP peptides induce the formation of extremely ordered, striated domains in supported DPPC bilayers. In this study, we show that WALP analogues with other uncharged residues (tyrosine, phenylalanine, or histidine at pH 9) can also induce the formation of striated domains, albeit in some cases with a slightly different pattern. The WALP analogues with positively charged residues (lysine or histidine at low pH) cannot induce striated domains and give rise to a completely different morphology: they induce irregularly shaped depressions in DPPC bilayers. The latter morphology is explained by the fact that the positively charged peptides repel each other and hence are not able to form striated domains in which they would have to be in close vicinity. They would reside in disordered, fluidlike lipid areas, appearing below the level of the ordered gel state lipid domains, which would account for the irregularly shaped depressions. PMID- 11851430 TI - A novel epsilon-cleavage within the transmembrane domain of the Alzheimer amyloid precursor protein demonstrates homology with Notch processing. AB - Proteolytic processing of the transmembrane domain of the amyloid precursor protein (APP) is a key component of Alzheimer's disease pathogenesis. Using C terminally tagged APP derivatives, we have identified by amino-terminal sequencing a novel cleavage site of APP, at Leu-49, distal to the gamma-secretase site. This was termed -cleavage. Brefeldin A treatment and pulse-chase experiments indicate that this cleavage occurs late in the secretory pathway. The level of -cleavage is decreased by expression of presenilin-1 mutants known to impair Abeta formation, and it is sensitive to the gamma-secretase inhibitors MDL28170 and L-685,458. Remarkably, it shares similarities with site 3 cleavage of Notch-1: membrane topology, cleavage before a valine, dependence on presenilins, and inhibition profile. PMID- 11851431 TI - Binding of ricin A-chain to negatively charged phospholipid vesicles leads to protein structural changes and destabilizes the lipid bilayer. AB - Ricin is a heterodimeric protein toxin in which a catalytic polypeptide (the A chain or RTA) is linked by a disulfide bond to a cell-binding polypeptide (the B chain or RTB). During cell entry, ricin undergoes retrograde vesicular transport to reach the endoplasmic reticulum (ER) lumen, from where RTA translocates into the cytosol, probably by masquerading as a substrate for the ER-associated protein degradation (ERAD) pathway. In partitioning studies in Triton X-114 solution, RTA is predominantly found in the detergent phase, whereas ricin holotoxin, native RTB, and several single-chain ribosome-inactivating proteins (RIPs) are in the aqueous phase. Fluorescence spectroscopy and far-UV circular dichroism (CD) demonstrated significant structural changes in RTA as a result of its interaction with liposomes containing negatively charged phospholipid (POPG). These lipid-induced structural changes markedly increased the trypsin sensitivity of RTA and, on the basis of the protein fluorescence determinations, abolished its ability to bind to adenine, the product resulting from RTA-catalyzed depurination of 28S ribosomal RNA. RTA also released trapped calcein from POPG vesicles, indicating that it destabilized the lipid bilayer. We speculate that membrane-induced partial unfolding of RTA during cell entry may facilitate its recognition as an ERAD substrate. PMID- 11851432 TI - Gene expression, mutation, and structure-function relationship of scorpion toxin BmP05 active on SK(Ca) channels. AB - Four peptide inhibitors of small-conductance Ca(2+)-activated, apamin-sensitive K(+) channels (SK(Ca)) have been isolated from the venom of the Chinese scorpion Buthus martensi, named BmP01, BmP02, BmP03, and BmP05, respectively [Romi-Lebrun, R. (1997) Eur. J. Biochem. 245, 457-464]. Among them BmP05 with 31 amino acid residues has been intensively studied due to its most potent toxicity. To investigate the structure-function relationship of BmP05, its wild type and seven mutants (their C-termini unamidated) were successfully expressed in the yeast secretion system and purified with a high yield over 8 mg/L. Their toxicity to mice and electrophysiological activity on the K(+) currents (SK(Ca) and Kv) in rat adrenal chromaffin cells were measured and compared. The results indicated the following: (1) As a selective antagonist against SK(Ca), 1 microM rBmP05 is equivalent to 0.2 microM apamin, and its IC(50) is 0.92 microM. (2) The basic residues Lys and Arg located at positions 6 and 13 in the N-terminal alpha-helix region are essential and synergetic in the interaction of the toxin with SK(Ca). (3) Disruption of the alpha-helix by mutation of Gln at position 9 with Pro results in almost total loss of toxicity. (4) The C-terminal residue His31 plays an auxiliary role in the interaction of the toxin with SK(Ca). (5) The beta-turn connecting two beta-sheets near the C-terminal part is responsible for the specificity of the toxin to the different subtypes of K(+) channels. PMID- 11851433 TI - Calnexin deficiency and endoplasmic reticulum stress-induced apoptosis. AB - In this study, we used calnexin-deficient cells to investigate the role of this protein in ER stress-induced apoptosis. We found that calnexin-deficient cells are relatively resistant to ER stress-induced apoptosis. However, caspase 3 and 8 cleavage and cytochrome c release were unchanged in these cells, indicating that ER to mitochondria "communication" during apoptotic stimulation is not affected in the absence of calnexin. The Bcl-2:Bax ratio was also not significantly changed in calnexin-deficient cells regardless of whether the ER stress was induced with thapsigargin or not. Ca(2+) homeostasis and ER morphology were unaffected by the lack of calnexin, but ER stress-induced Bap31 cleavage was significantly inhibited. Immunoprecipitation experiments revealed that Bap31 forms complexes with calnexin, which may play a role in apoptosis. The results suggest that calnexin may not play a role in the initiation of the ER stress but that the protein has an effect on later apoptotic events via its influence on Bap31 function. PMID- 11851434 TI - Thrombin hydrolysis of V29F and V34L mutants of factor XIII (28-41) reveals roles of the P(9) and P(4) positions in factor XIII activation. AB - In blood coagulation, thrombin helps to activate factor XIII by cleaving the activation peptide at the R37-G38 peptide bond. The more easily activated factor XIII V34L has been correlated with protection from myocardial infarction. V34L and V29F factor XIII mutant peptides were designed to further characterize substrate binding to thrombin. HPLC kinetic studies have been carried out on thrombin hydrolysis of FXIII activation peptide (28-41), FXIII (28-41) V34L, FXIII (28-41) V29F, and FXIII (28-41) V29F V34L. The V34L mutations lead to improvements in both K(m) and k(cat) whereas the V29F mutation primarily affects K(m). Interactions of the peptides with thrombin have been monitored by 1D proton line broadening NMR and 2D transferred NOESY studies. The results were compared with previously published X-ray crystal structures of thrombin-bound fibrinogen Aalpha (7-16), thrombin receptor PAR1 (38-60), and factor XIII (28-37). In solution, the (34)VVPR(37) and (34)LVPR(37) segments of the factor XIII activation peptide serve as the major anchor points onto thrombin. The N-terminal segments are proposed to interact transiently with the enzyme surface. Long-range NOEs from FXIII V29 or F29 toward (34)V/LVPR(37) have not been observed by NMR studies. Overall, the kinetic and NMR results suggest that the factor XIII activation peptide binds to thrombin in a manner more similar to the thrombin receptor PAR1 than to fibrinogen Aalpha. The V29 and V34 positions affect, in different ways, the ability of thrombin to effectively hydrolyze the activation peptide. Mutations at these sites may prove useful in controlling factor XIII activation. PMID- 11851435 TI - Evidence for a global inhibitor-induced conformation change on the Ca(2+)-ATPase of sarcoplasmic reticulum from paired inhibitor studies. AB - The Ca(2+)-ATPase of skeletal muscle sarcoplasmic reticulum is inhibited by a variety of hydrophobic, hydroxy-containing molecules. A kinetic method has been used to study competition between binding of pairs of inhibitors to the ATPase. The presence of 2,5-di-tert-butyl-1,4-dihydroxybenzene (BHQ) decreases the affinity of the ATPase for 2,5-dipropyl-1,4-dihydroxybenzene (PHQ), suggesting that PHQ and BHQ bind to the same site on the ATPase. In contrast, the presence of BHQ increases the affinity of the ATPase for curcumin and vice versa. This suggests that BHQ and curcumin bind to separate sites on the ATPase and that binding of the first inhibitor to the ATPase results in a change to a conformation with higher affinity for the second inhibitor. This is consistent with previous experiments with BHQ and thapsigargin suggesting a conformation change on inhibitor binding, E2 + I <--> 2; E2I <--> 2; E2(A)I, with E2(A)I having a higher affinity for the second inhibitor than E2. The affinity for BHQ is also increased by binding of diethylstilbesterol, ellagic acid, or nonylphenol, and the affinity for curcumin is also increased by ellagic acid. These results showing that binding of a variety of inhibitors of very different structures all result in a general increase in inhibitor affinity point to a global conformational change on the Ca(2+)-ATPase caused by inhibitor binding, as well as any local, inhibitor-specific changes in conformation. PMID- 11851438 TI - Introduction: Force and Tunneling Microscopy. PMID- 11851436 TI - Conformation of fork junction DNA in a complex with Escherichia coli RNA polymerase. AB - Escherichia coli RNA polymerase is able to bind fork junction DNA containing a conserved -10 promoter element in a sequence-specific manner, and it is believed that polymerase-fork junction DNA interaction mimics those between the enzyme and the promoter DNA in the open complex. In this report we determined the conformation of polymerase-bound fork junction DNA in solution. A series of distances between sites in the fork junction DNA in complex with polymerase were determined using luminescence and fluorescence resonance energy transfer. A series of fork junction DNAs were prepared containing the luminescent or fluorescent donor probe at the upstream or at the downstream end of the fork DNA and acceptor probes at nine positions within the fork junction DNA. The measured distances were compared with analogous distances in a model reference DNA duplex, and the observed distance differences were used to build a model of the fork junction DNA in a complex with the polymerase. The obtained model revealed an insignificant perturbation of the duplex part of the fork DNA in a complex with the polymerase whereas a sharp kink of DNA was observed at the ds/ss DNA boundary of the fork junction DNA. PMID- 11851439 TI - Cross-Sectional Scanning Tunneling Microscopy. PMID- 11851440 TI - Effect of Strain on Structure and Morphology of Ultrathin Ge Films on Si(001). PMID- 11851441 TI - Scanning Tunneling Microscopy Studies of Metal on Metal Epitaxy. PMID- 11851442 TI - Scanning Tunneling Microscopy Imaging of Small Adsorbed Molecules on Metal Surfaces in an Ultrahigh Vacuum Environment. PMID- 11851443 TI - Images of Adsorbates with the Scanning Tunneling Microscope: Theoretical Approaches to the Contrast Mechanism. PMID- 11851444 TI - Characterization of Organosulfur Molecular Monolayers on Au(111) using Scanning Tunneling Microscopy. PMID- 11851445 TI - Electrochemical Applications of in Situ Scanning Probe Microscopy. PMID- 11851446 TI - Scratching the Surface: Fundamental Investigations of Tribology with Atomic Force Microscopy. PMID- 11851447 TI - Nanometer-Scale Surface Modification Using the Scanning Probe Microscope: Progress since 1991. PMID- 11851449 TI - Proteinminus signProtein Interactions: Interface Structure, Binding Thermodynamics, and Mutational Analysis. PMID- 11851448 TI - Introduction: Molecular Recognition. PMID- 11851450 TI - Protein Structure and the Energetics of Protein Stability. PMID- 11851451 TI - Nearest-Neighbor Recognition in Phospholipid Membranes. PMID- 11851452 TI - Binding Energy and Catalysis: The Implications for Transition-State Analogs and Catalytic Antibodies. PMID- 11851454 TI - The Stability of Cyclodextrin Complexes in Solution. PMID- 11851453 TI - The Cationminus signpi Interaction. PMID- 11851455 TI - Molecular Recognition of Proteinminus signLigand Complexes: Applications to Drug Design. PMID- 11851456 TI - Preorganization of DNA: Design Principles for Improving Nucleic Acid Recognition by Synthetic Oligonucleotides. PMID- 11851457 TI - A Structural Basis for RNAminus signLigand Interactions. PMID- 11851458 TI - Signaling Recognition Events with Fluorescent Sensors and Switches. PMID- 11851459 TI - Steroids in Molecular Recognition. PMID- 11851460 TI - Artificial Organic Host Molecules for Anions. PMID- 11851461 TI - Self-Assembling Capsules. PMID- 11851462 TI - Formation of Artificial Receptors by Metal-Templated Self-Assembly. PMID- 11851463 TI - Dendrimers in Supramolecular Chemistry: From Molecular Recognition to Self Assembly. PMID- 11851464 TI - Novel Cavity Design Using Calix[n]arene Skeletons: Toward Molecular Recognition and Metal Binding. PMID- 11851465 TI - Introduction: Polyketide and Nonribosomal Polypeptide Biosynthesis. From Collie to Coli. PMID- 11851466 TI - Genetic Contributions to Understanding Polyketide Synthases. PMID- 11851467 TI - Biosynthesis of the Kinamycins. PMID- 11851468 TI - Anthracycline Biosynthesis in Streptomyces galilaeus. PMID- 11851469 TI - Biosynthetic Studies of Daunorubicin and Tetracenomycin C. PMID- 11851470 TI - Enzymology and Molecular Biology of Aflatoxin Biosynthesis. PMID- 11851471 TI - Manipulation of Modular Polyketide Synthases. PMID- 11851472 TI - Harnessing the Biosynthetic Potential of Modular Polyketide Synthases. PMID- 11851473 TI - Avermectin Biosynthesis. PMID- 11851474 TI - Biosynthesis of Erythromycin and Rapamycin. PMID- 11851475 TI - The Mechanism of ACV Synthetase. PMID- 11851476 TI - Modular Peptide Synthetases Involved in Nonribosomal Peptide Synthesis. PMID- 11851477 TI - Multifunctional Peptide Synthetases. PMID- 11851478 TI - Vanadium in Modern Organic Synthesis. PMID- 11851479 TI - Bona Fide Predictions of Protein Secondary Structure Using Transparent Analyses of Multiple Sequence Alignments. PMID- 11851480 TI - Structure and Dynamics of Zeolites Investigated by Molecular Dynamics. PMID- 11851482 TI - Rotational Tunneling and Neutron Spectroscopy: A Compilation. PMID- 11851481 TI - Activation of Cminus signH Bonds by Metal Complexes. PMID- 11851483 TI - Developments in Measuring and Calculating Chemical Vapor Transport Phenomena Demonstrated on Cr, Mo, W, and Their Compounds. PMID- 11851484 TI - Comparative QSAR: Radical Reactions of Benzene Derivatives in Chemistry and Biology. PMID- 11851485 TI - Electrochemical Carbon. PMID- 11851486 TI - Carrier-Based Ion-Selective Electrodes and Bulk Optodes. 1. General Characteristics. PMID- 11851487 TI - Low Molecular Mass Gelators of Organic Liquids and the Properties of Their Gels. PMID- 11851488 TI - trans-1,2-Diaminocyclohexane Derivatives as Chiral Reagents, Scaffolds, and Ligands for Catalysis: Applications in Asymmetric Synthesis and Molecular Recognition. PMID- 11851489 TI - Rhenium(VII) Oxo and Imido Complexes: Synthesis, Structures, and Applications. PMID- 11851490 TI - Alkylphenol Ethoxylates: Trace Analysis and Environmental Behavior. PMID- 11851491 TI - Chemistry of beta-(Acyloxy)alkyl and beta-(Phosphatoxy)alkyl Radicals and Related Species: Radical and Radical Ionic Migrations and Fragmentations of Carbonminus signOxygen Bonds. PMID- 11851492 TI - Enantiomeric Recognition of Amine Compounds by Chiral Macrocyclic Receptors. PMID- 11851493 TI - The Coordination Chemistry of the CF Unit in Fluorocarbons. PMID- 11851494 TI - Lipophilicity Profiles of Ampholytes. PMID- 11851495 TI - Synthetic Applications of Dialkyl (Chloromethyl)phosphonates and N,N,N',N' Tetraalkyl(chloromethyl)phosphonic Diamides. PMID- 11851496 TI - Organometallic Fluorides: Compounds Containing Carbonminus signMetalminus signFluorine Fragments of d-Block Metals. PMID- 11851497 TI - Introduction: Polyoxometalates-Multicomponent Molecular Vehicles To Probe Fundamental Issues and Practical Problems. PMID- 11851499 TI - The Nomenclature of Polyoxometalates: How To Connect a Name and a Structure. PMID- 11851498 TI - Present General Status of Understanding of Heteropoly Electrolytes and a Tracing of Some Major Highlights in the History of Their Elucidation. PMID- 11851500 TI - Main-Group Element, Organic, and Organometallic Derivatives of Polyoxometalates. PMID- 11851501 TI - Homogeneous-Phase Electron-Transfer Reactions of Polyoxometalates. PMID- 11851502 TI - Catalysis by Heteropoly Acids and Multicomponent Polyoxometalates in Liquid-Phase Reactions. PMID- 11851503 TI - Heterogeneous Catalysis. PMID- 11851504 TI - Electrochemical Properties of Polyoxometalates as Electrocatalysts. PMID- 11851505 TI - Polyoxometalates: Very Large Clusters-Nanoscale Magnets. PMID- 11851506 TI - Polyoxometalate-Based Molecular Materials. PMID- 11851507 TI - Polyoxoanion Chemistry Moves toward the Future: From Solids and Solutions to Surfaces. PMID- 11851508 TI - Photo- and Electrochromism of Polyoxometalates and Related Materials. PMID- 11851509 TI - Polyoxometalates in Medicine. PMID- 11851510 TI - A Survey of Applications of Polyoxometalates. PMID- 11851511 TI - Homogeneous-Phase Electron-Transfer Reactions of Polyoxometalates.[(Chem. Rev. 1998, 98, 113 (this issue). Published on the Web January 22, 1998.]. PMID- 11851514 TI - Preventing type 1 diabetes may someday become possible. PMID- 11851515 TI - Experimental vaccine for recurrent UTIs. PMID- 11851516 TI - Disparity in cancer statistics changing. PMID- 11851517 TI - "Sobering" levels of drug-resistant HIV found. PMID- 11851522 TI - Predictive value of the electrocardiogram in acute myocardial infarction. PMID- 11851524 TI - Is the size of low-density lipoprotein particles related to the risk of coronary heart disease? PMID- 11851526 TI - Sequential assessment of multiple organ dysfunction as a predictor of outcome. PMID- 11851528 TI - Drug-related hyperglycemia. PMID- 11851529 TI - Drug-related hyperglycemia. PMID- 11851531 TI - Reforming graduate medical education. PMID- 11851532 TI - Identifying genetic factors in Parkinson disease. PMID- 11851534 TI - Treatment of type 2 diabetes mellitus in children. PMID- 11851536 TI - How much oversight is necessary to protect human subjects? PMID- 11851538 TI - Cardiovascular effects of sildenafil during exercise in men with known or probable coronary artery disease: a randomized crossover trial. AB - CONTEXT: The relationship between sildenafil citrate use and reported adverse cardiovascular events in men with coronary artery disease (CAD) is unclear. OBJECTIVE: To evaluate the cardiovascular effects of sildenafil during exercise in men with CAD. DESIGN, SETTING, AND SUBJECTS: Randomized, double-blind, placebo controlled crossover trial conducted March to October 2000 at a US ambulatory care referral center among 105 men with a mean (SD) age of 66 (9) years who had erectile dysfunction and known or highly suspected CAD. INTERVENTIONS: All patients underwent 2 symptom-limited supine bicycle echocardiograms separated by an interval of 1 to 3 days after receiving a single dose of sildenafil (50 or 100 mg) or placebo 1 hour before each exercise test. MAIN OUTCOME MEASURES: Hemodynamic effects of sildenafil during exercise (onset, extent, and severity of ischemia) assessed by exercise echocardiography. RESULTS: Mean (SD) resting ejection fraction was 56% (7%) (range, 39%-68%). After sildenafil use, resting systolic blood pressure was reduced from 135 (19) mm Hg to 128 (17) mm Hg, for a mean change of -7 mm Hg (95% confidence interval [CI], -9 to -4 mm Hg; P<.001). After placebo use, the mean (SD) change was from 135 (20) mm Hg to 133 (19) mm Hg, a difference of -2 mm Hg (95% CI, -6 to 0.3 mm Hg; P =.08). The difference between mean change after sildenafil and placebo use was 4.3 (95% CI, 0.9-7.7; P =.01). Resting heart rate, diastolic blood pressure, and wall motion score index (a measure of the extent and severity of wall motion abnormalities) did not change significantly in either group. Exercise capacity was similar with sildenafil use (mean [SD], 4.5 [1.0] metabolic equivalents) and placebo use (mean [SD], 4.6 [1.0] metabolic equivalents; mean difference, 0.07; 95% CI, -.06 to 0.19; P =.29). Exercise blood pressure and heart rate increments were similar. Dyspnea or angina developed in 69 patients who took sildenafil and 70 patients who took placebo (P =.89); exercise electrocardiography was positive in 12 patients (11%) who took sildenafil and 17 patients (16%) who took placebo (P =.09). Exercise-induced wall motion abnormalities developed in similar numbers of patients after sildenafil and placebo use (84 and 86 patients, respectively; P =.53). Wall motion score index at peak exercise was similar after sildenafil and placebo use (mean [SD], 1.4 [0.4] vs 1.4 [0.4]; mean difference, 0.01; 95% CI, 0.01 to 0.03; P =.40). CONCLUSION: In men with stable CAD, sildenafil had no effect on symptoms, exercise duration, or presence or extent of exercise-induced ischemia, as assessed by exercise echocardiography. PMID- 11851539 TI - Untreated gonococcal and chlamydial infection in a probability sample of adults. AB - CONTEXT: The prevalence and distribution of gonococcal and chlamydial infections in the general population are poorly understood. Development of nucleic acid amplification tests, such as the ligase chain reaction assay, provides new opportunities to estimate the prevalence of untreated infections in the population. OBJECTIVE: To estimate the overall prevalence of untreated gonococcal and chlamydial infections and to describe patterns of infection within specific demographic subgroups of the young adult population in Baltimore, Md. DESIGN AND SETTING: Cross-sectional behavioral survey based on a probability sample of Baltimore households with collection of urine specimens between January 1997 and September 1998. PARTICIPANTS: A total of 728 adults aged 18 to 35 years completed the interview portion of the study, and 579 of these respondents also provided a urine specimen adequate for testing. MAIN OUTCOME MEASURE: Prevalence of untreated infection, as measured by the percentage of specimens testing positive for gonococcal and chlamydial infection by ligase chain reaction, weighted to reflect variations in probabilities of sample selection from the population. Alternate estimates of the prevalence of recent treated infection were derived from clinically diagnosed cases reported to the Baltimore City Health Department and by diagnoses reported by participants in the survey. RESULTS: An estimated 5.3% (SE, 1.4%) of the population aged 18 to 35 years has an untreated gonococcal infection, and 3.0% (SE, 0.8%) is estimated to have an untreated chlamydial infection. While 7.9% (SE, 1.6%) of the population is estimated to have either an untreated gonococcal or chlamydial infection, estimated prevalence is substantially higher among black women (15.0%; SE, 3.7%). Few participants with untreated infections reported dysuria or discharge during the 6 months preceding testing. The estimated number of untreated gonococcal infections in the population (9241; SE, 2441) substantially exceeds both the number of such infections diagnosed among Baltimore adults aged 18 to 35 years and reported to the Baltimore City Health Department during 1998 (4566), and the estimated number of diagnoses derived using participants' reports for the 12 months prior to the survey (4708 [SE, 1918] to 5231 [SE, 2092]). The estimated number of untreated chlamydial infections (5231; SE, 1395) is also greater than the number of cases reported to the health department in 1998 (3664) but is slightly less than the estimated number of diagnoses derived using participants' reports of chlamydial infections diagnosed during the 12 months prior to the survey (5580 [SE, 1918] to 6975 [SE, 2441]). CONCLUSION: In 1997-1998, the estimated number of undiagnosed gonococcal and chlamydial infections prevalent in the population of Baltimore adults aged 18 to 35 years approached or exceeded the number of infections that were diagnosed and treated annually. PMID- 11851540 TI - Hormone replacement therapy in relation to breast cancer. AB - CONTEXT: Studies of long-term hormone replacement therapy (HRT) suggest an associated increased risk of breast cancer, but whether this association differs according to histologic type of cancer has not been extensively studied. OBJECTIVE: To determine whether the association between HRT and risk of breast cancer varies by HRT formulation and differs across histologic cancer types. DESIGN, SETTING, AND PARTICIPANTS: Nested case-control study among 705 postmenopausal women enrolled in the Group Health Cooperative of Puget Sound (GHC) who were aged 50 to 74 years and had primary invasive breast cancer diagnosed between July 1, 1990, and December 31, 1995 (cases), and 692 randomly selected aged-matched female members of GHC (controls). MAIN OUTCOME MEASURE: Incidence and type of breast cancer by duration of HRT use in the 5-year period ending 1 year before diagnosis, which was ascertained from computerized pharmacy records. RESULTS: The incidence of breast cancer, all histologic types combined, was increased by 60% to 85% in recent long-term users of HRT, whether estrogen alone or estrogen plus progestin. Longer use of HRT (odds ratio [OR], 3.07 for 57 months or more; 95% confidence interval [CI], 1.55-6.06) and current use of combination therapy (OR, 3.91; 95% CI, 2.05-7.44) were associated with increased risk of lobular breast cancer. Long-term HRT use was associated with a 50% increase in nonlobular cancer (OR, 1.52 for 57 months or more; 95% CI, 1.01 2.29). CONCLUSION: Our data add to the growing body of evidence that recent long term use of HRT is associated with an increased risk of breast cancer and that such use may be related particularly to lobular tumors. PMID- 11851541 TI - Participation in cognitively stimulating activities and risk of incident Alzheimer disease. AB - CONTEXT: Frequent participation in cognitively stimulating activities has been hypothesized to reduce risk of Alzheimer disease (AD), but prospective data regarding an association are lacking. OBJECTIVE: To test the hypothesis that frequent participation in cognitive activities is associated with a reduced risk of AD. DESIGN: Longitudinal cohort study with baseline evaluations performed between January 1994 and July 2001 and mean follow-up of 4.5 years. PARTICIPANTS AND SETTING: A total of 801 older Catholic nuns, priests, and brothers without dementia at enrollment, recruited from 40 groups across the United States. At baseline, they rated frequency of participation in common cognitive activities (eg, reading a newspaper), from which a previously validated composite measure of cognitive activity frequency was derived. MAIN OUTCOME MEASURES: Clinical diagnosis of AD by a board-certified neurologist using National Institute of Neurological and Communicative Disorders and Stroke/Alzheimer's Disease and Related Disorders Association criteria and change in global and specific measures of cognitive function, compared by cognitive activity score at baseline. RESULTS: Baseline scores on the composite measure of cognitive activity ranged from 1.57 to 4.71 (mean, 3.57; SD, 0.55), with higher scores indicating more frequent activity. During an average of 4.5 years of follow-up, 111 persons developed AD. In a proportional hazards model that controlled for age, sex, and education, a 1 point increase in cognitive activity score was associated with a 33% reduction in risk of AD (hazard ratio, 0.67; 95% confidence interval, 0.49-0.92). Results were comparable when persons with memory impairment at baseline were excluded and when terms for the apolipoprotein E epsilon4 allele and other medical conditions were added. In random-effects models that controlled for age, sex, education, and baseline level of cognitive function, a 1-point increase in cognitive activity was associated with reduced decline in global cognition (by 47%), working memory (by 60%), and perceptual speed (by 30%). CONCLUSION: These results suggest that frequent participation in cognitively stimulating activities is associated with reduced risk of AD. PMID- 11851542 TI - Discussing religious and spiritual issues at the end of life: a practical guide for physicians. AB - As patients near the end of life, their spiritual and religious concerns may be awakened or intensified. Many physicians, however, feel unskilled and uncomfortable discussing these concerns. This article suggests how physicians might respond when patients or families raise such concerns. First, some patients may explicitly base decisions about life-sustaining interventions on their spiritual or religious beliefs. Physicians need to explore those beliefs to help patients think through their preferences regarding specific interventions. Second, other patients may not bring up spiritual or religious concerns but are troubled by them. Physicians should identify such concerns and listen to them empathetically, without trying to alleviate the patient's spiritual suffering or offering premature reassurance. Third, some patients or families may have religious reasons for insisting on life-sustaining interventions that physicians advise against. The physician should listen and try to understand the patient's viewpoint. Listening respectfully does not require the physician to agree with the patient or misrepresent his or her own views. Patients and families who feel that the physician understands them and cares about them may be more willing to consider the physician's views on prognosis and treatment. By responding to patients' spiritual and religious concerns and needs, physicians may help them find comfort and closure near the end of life. PMID- 11851543 TI - Virus-induced asthma attacks. AB - Viral respiratory tract infections are a common cause of asthma attacks. Study of this phenomenon has revealed multiple mechanisms and contributed to understanding of the increase in airway inflammation and bronchoconstriction observed in this context. Changes in the neural control of the airways contribute to bronchoconstriction, which is reflected in an increased efficacy of anticholinergic medications during acute asthma attacks. The ability to prevent or treat viral respiratory tract infections is currently limited. However, as more effective antiviral treatments and vaccines become available, such therapies are likely to be effective in patients with asthma. Clinical management of this problem is illustrated in this article by the case of a 40-year-old woman with history of mild asthma who was admitted to an intensive care unit with severe bronchospasm and an upper respiratory tract infection. PMID- 11851544 TI - Postpartum depression. PMID- 11851545 TI - Safe sex for men with coronary artery disease: exercise, sildenafil, and risk of cardiac events. PMID- 11851546 TI - Unveiling the hidden epidemic of sexually transmitted diseases. PMID- 11851548 TI - MSJAMA. Mass media and medicine: challenges and opportunities. PMID- 11851549 TI - The media and public reaction to genetic research. PMID- 11851550 TI - Marketing medicine to the public: a reader's guide. PMID- 11851551 TI - Journalists as change agents in medicine and health care. PMID- 11851558 TI - Genome analysis yields mutations linked to hereditary prostate cancer. PMID- 11851559 TI - Strep A, neuropsychiatric disorders tie found. PMID- 11851560 TI - Returning astronaut study helps some reeling patients. PMID- 11851565 TI - Safety of implantable cardiac devices. PMID- 11851568 TI - Do patents prevent access to drugs for HIV in developing countries? PMID- 11851569 TI - Do patents prevent access to drugs for HIV in developing countries? PMID- 11851570 TI - Do patents prevent access to drugs for HIV in developing countries? PMID- 11851572 TI - Research as a quality improvement activity. PMID- 11851574 TI - Pharmaceutical representatives and resident physicians. PMID- 11851575 TI - Effects of hydration on fluid balance and lower-extremity blood viscosity during long airplane flights. PMID- 11851576 TI - Raloxifene and cardiovascular events in osteoporotic postmenopausal women: four year results from the MORE (Multiple Outcomes of Raloxifene Evaluation) randomized trial. AB - CONTEXT: Raloxifene, a selective estrogen receptor modulator, improves cardiovascular risk factors, but its effect on cardiovascular events is unknown. OBJECTIVE: To determine the effect of raloxifene on cardiovascular events in osteoporotic postmenopausal women. DESIGN: Secondary analysis of data from the Multiple Outcomes of Raloxifene Evaluation trial, a randomized, double-blind, placebo-controlled trial conducted between November 1994 and September 1999. SETTING: Outpatient and community settings at 180 sites in 25 countries. PARTICIPANTS: A total of 7705 osteoporotic postmenopausal women (mean age, 67 years). INTERVENTION: Patients were randomly assigned to receive raloxifene, 60 mg/d (n = 2557), or 120 mg/d (n = 2572), or placebo (n = 2576) for 4 years. MAIN OUTCOME MEASURES: Cardiovascular events, including coronary events (myocardial infarction, unstable angina, or coronary ischemia) and cerebrovascular events (stroke or transient ischemic attack), collected as safety end points and subsequently adjudicated by a cardiologist blinded to therapy. Cardiovascular risk at study entry was determined by the presence of multiple cardiovascular risk factors or prior coronary events or revascularization procedure. RESULTS: In the overall cohort, there were no significant differences between treatment groups in the number of combined coronary and cerebrovascular events: 96 (3.7%) with placebo, 82 (3.2%) with 60 mg/d of raloxifene, and 94 (3.7%) with 120 mg/d of raloxifene. Relative risks (RRs) were 0.86 (95% confidence interval [CI], 0.64 1.15) and 0.98 (95% CI, 0.74-1.30) for 60 mg/d and 120 mg/d of raloxifene, respectively. Similar results were obtained when coronary and cerebrovascular events were analyzed separately. Among the subset of 1035 women with increased cardiovascular risk at baseline, those assigned to raloxifene had a significantly lower risk of cardiovascular events compared with placebo (RR, 0.60; 95% CI, 0.38 0.95 for both raloxifene groups). The number of cardiovascular events during the first year was not significantly different across groups in the overall cohort (P =.94), or among women at increased cardiovascular risk (P =.86) or with evidence of established coronary heart disease (P =.60). CONCLUSIONS: Raloxifene therapy for 4 years did not significantly affect the risk of cardiovascular events in the overall cohort but did significantly reduce the risk of cardiovascular events in the subset of women with increased cardiovascular risk. There was no evidence that raloxifene caused an early increase in risk of cardiovascular events. Before raloxifene is used for prevention of cardiovascular events, these findings require confirmation in trials with evaluation of cardiovascular outcomes as the primary objective. PMID- 11851577 TI - Fatal inhalational anthrax with unknown source of exposure in a 61-year-old woman in New York City. AB - A 61-year-old woman who was a New York City hospital employee developed fatal inhalational anthrax, but with an unknown source of anthrax exposure. The patient presented with shortness of breath, malaise, and cough that had developed 3 days prior to admission. Within hours of presentation, she developed respiratory failure and septic shock and required mechanical ventilation and vasopressor therapy. Spiral contrast-enhanced computed tomography of the chest demonstrated large bilateral pleural effusions and hemorrhagic mediastinitis. Blood cultures, as well as DNA amplification by polymerase chain reaction of the blood, bronchial washings, and pleural fluid specimens, were positive for Bacillus anthracis. The clinical course was complicated by liver failure, renal failure, severe metabolic acidosis, disseminated intravascular coagulopathy, and cardiac tamponade, and the patient died on the fourth hospital day. The cause of death was inhalational anthrax. Despite epidemiologic investigation, including environmental samples from the patient's residence and workplace, no mechanism for anthrax exposure has been identified. PMID- 11851578 TI - Fatal inhalational anthrax in a 94-year-old Connecticut woman. AB - We describe the 11th case of bioterrorism-related inhalational anthrax reported in the United States. The presenting clinical features of this 94-year-old woman were subtle and nondistinctive. The diagnosis was recognized because blood cultures were obtained prior to administration of antibiotics, emphasizing the importance of this diagnostic test in evaluating ill patients who have been exposed to Bacillus anthracis. The patient's clinical course was characterized by progression of respiratory insufficiency, pleural effusions and pulmonary edema, and, ultimately, death. Although her B anthracis bacteremia was rapidly sterilized after initiation of antibiotic therapy, viable B anthracis was present in postmortem mediastinal lymph node specimens. The source of exposure to B anthracis in this patient is not known. Exposure to mail that was cross contaminated as it passed through postal facilities contaminated with B anthracis spores is one hypothesis under investigation. PMID- 11851580 TI - Secondary and tertiary palliative care in US hospitals. AB - Palliative care services provide secondary and tertiary levels of palliative care, the interdisciplinary care of patients in which the goal is comfort and quality of life. Primary palliative care refers to the basic skills and competencies required of all physicians and other health care professionals. Secondary palliative care refers to the specialist clinicians and organizations that provide consultation and specialty care. Tertiary palliative care refers to the academic medical centers where specialist knowledge for the most complex cases is practiced, researched, and taught. The case of Reverend J, a man with advanced cancer admitted to an acute palliative care unit in a teaching hospital, illustrates the use of secondary and tertiary clinical palliative care services in hospitals and health care systems. PMID- 11851579 TI - Cutaneous anthrax associated with microangiopathic hemolytic anemia and coagulopathy in a 7-month-old infant. AB - A 7-month-old infant with cutaneous anthrax developed severe systemic illness despite early treatment with antibiotics. The infant displayed severe microangiopathic hemolytic anemia with renal involvement, coagulopathy, and hyponatremia. These findings are unusual with cutaneous anthrax, but have been described in illness resulting from spider toxin and may delay correct diagnosis. The systemic manifestations of the disease persisted for nearly a month despite corticosteroid therapy, but resolved. PMID- 11851581 TI - Caring for bereaved patients: "All the doctors just suddenly go". PMID- 11851582 TI - beta-Blocker therapy in heart failure: scientific review. AB - CONTEXT: Care of patients with heart failure has been revolutionized throughout the past decade. A paradigm shift in the strategy for treating heart failure caused by systolic dysfunction is in progress. Despite the initial perception about beta-blockers' safety, they are now the most extensively studied class of agents in the treatment of heart failure and have emerged as an important intervention to improve the clinical outcomes of heart failure patients. OBJECTIVE: To provide scientific rationale for the use of beta-blockers for patients with heart failure. DATA SOURCES: All English-language articles of large, randomized controlled clinical trials assessing the mortality benefits of beta-blockers in patients with heart failure were identified to provide the scientific rationale for the use of beta-blockers in heart failure. Basic science studies were reviewed to provide an overview of the potential physiologic role of beta-blockers in heart failure. Finally, clinical guidelines for the treatment of patients with heart failure were assessed to determine current recommendations for the use of these agents. STUDY SELECTION AND DATA EXTRACTION: Randomized controlled clinical trials of beta-blockers that included more than 300 subjects and assessed mortality as a primary end point. DATA SYNTHESIS: Of the 4 beta blockers tested in large randomized controlled clinical trials of patients with heart failure, 3 are available in the United States, bisoprolol, carvedilol, and metoprolol; 2 of these, carvedilol and metoprolol, have Food and Drug Administration indications for the treatment of heart failure. Compared with placebo treatment, beta-blocker use is associated with a consistent 30% reduction in mortality and a 40% reduction in hospitalizations in patients with class II and III heart failure. CONCLUSIONS: Tested in more than 10,000 patients, beta blockers reduce morbidity and mortality in class II through IV heart failure. Along with angiotensin-converting enzyme inhibitors, digoxin, and diuretics, beta blockers have strengthened the armamentarium to improve clinical outcomes of heart failure patients. The science supporting beta-blockers must be translated into practice safely and rationally if the agents are to achieve their full potential. PMID- 11851583 TI - beta-Blockers in heart failure: clinical applications. AB - beta-Blockers reduce morbidity and mortality in heart failure patients with left ventricular systolic dysfunction and stable fluid status. The successful adoption of beta-blocker guidelines for these patients requires an understanding of the value of this therapy and effective systems to maintain safety and ensure high quality of care. This article distills scientific evidence and consensus guidelines into a series of cases and practical answers about patient selection, discussions with patients, management and monitoring, and systems improvements to optimize quality of care, safety, and benefit for all patients with heart failure. PMID- 11851584 TI - Bioterrorism preparedness and response: clinicians and public health agencies as essential partners. PMID- 11851591 TI - Effect of pH changes on reactivity of rat mesenteric artery segments at different magnitude of stretch. AB - The reaction to noradrenaline (NA) (10 microM) and electrical field stimulation (EFS) was studied in rat mesenteric artery segments at different magnitude of stretch and the solution pH. Alkaline solution (pH 7.8) potentiated and acidic solution (pH 7.0 or 6.6) inhibited the EFS-evoked response of segments stretched to values corresponding to arterial pressure of 5-200 mmHg. These pH changes failed to alter resting tension at any magnitude of stretch. Acedic solution of pH 6.6 caused 2-fold decrease in noradrenaline- and 5-15-fold decrease in the EFS evoked response of segments stretched to values corresponding to arterial pressure of 50, 125, and 200 mmHg. In segments pre-contracted with noradrenaline (10 microM) acidification caused the decrease of the dilation and appearance of the constriction induced by the EFS. The effect of acidosis on the EFS-evoked response was diminished and the effect on noradrenaline-evoked response was abolished in the presence of nitric oxide synthase blocker, NG-nitro-L-arginine (100 microM). These results suggest that acidosis effectively impairs reactivity of the rat mesenteric artery in a wide range of its stretch, and the inhibition of the response to noradrenaline occurs completely, while to EFS only partially due to nitric oxide (NO) release, presumably by the endothelium. In addition, it was shown that acidosis is able to act not only as the commonly known dilator agent, but also as an agent potentiating constriction in case of the high noradrenaline-induced tone. PMID- 11851592 TI - Modulation of vascular tone and reactivity by nitric oxide in porcine pulmonary arteries and veins. AB - Isolated porcine pulmonary vessels were studied in order to evaluate the role of nitric oxide in arteries and veins. Leukotriene C4 and noradrenaline contracted porcine pulmonary arteries but induced only negligible contractions of porcine pulmonary veins. After treatment with the nitric oxide synthase inhibitor N(omega)-nitro-L-arginine (L-NOARG), significant contractions to leukotriene C4 and noradrenaline were uncovered in pulmonary veins. In arterial preparations, L NOARG caused a less marked potentiation of noradrenaline-induced contractions and did not alter leukotriene C4-induced contractions. Endothelium-dependent relaxations to acetylcholine were greater in veins compared with arteries whereas the endothelium-independent relaxations to the nitric oxide donor sodium nitroprusside (SNP) and the cyclic nucleotide analogue 8-bromo-cGMP were similar in the two preparations. Taken together these data suggest that the apparent insensitivity of porcine pulmonary veins to leukotriene C4 and noradrenaline was because of release of nitric oxide. The effect of nitric oxide synthase inhibition was less pronounced in porcine pulmonary arteries, suggesting a preferential functional role of nitric oxide in porcine pulmonary veins, originating in a greater production of nitric oxide by veins as opposed to arteries. PMID- 11851593 TI - Mechanics of human triceps surae muscle in walking, running and jumping. AB - Length changes of the muscle-tendon complex (MTC) during activity are in part the result of length changes of the active muscle fibres, the contractile component (CC), and also in part the result of stretch of elastic structures [series elastic component (SEC)]. We used a force platform and kinematic measurements to determine force and length of the human calf muscle during walking, running and squat jumping. The force-length relation of the SEC was determined in dynamometer experiments on the same four subjects. Length of the CC was calculated as total muscle-tendon length minus the force dependent length of the SEC. The measured relations between force and length or velocity were compared with the individually determined force-length and force-velocity relations of the CC. In walking or running the negative work performed in the eccentric phase was completely stored as elastic energy. This elastic energy was released in the concentric phase, at speeds well exceeding the maximum shortening speed predicted by the Hill force-velocity relation. Speed of the CC, in contrast, was positive and low, well within the range predicted by the measured force-velocity properties and compatible with a favourable muscular efficiency. These effects were also present in purely concentric contractions, like the squatted jump. Contractile component length usually started at the far end of the force-length relation. Inter-individual differences in series-elastic stiffness were reflected in the force and length recordings during natural activity. PMID- 11851594 TI - Sweating response in physically trained men to sustained handgrip exercise in mildly hyperthermic conditions. AB - To investigate the effects of physical training on heat loss response to sustained handgrip exercise (non-thermal factors), we compared the sweating response during isometric handgrip exercise to mild hyperthermia in physically trained and untrained subjects. Seven trained and untrained male subjects (maximal oxygen uptake 62.7 +/- 2.4 and 42.7 +/- 1.6 mL kg-1 min-1, respectively, P < 0.05) performed isometric handgrip exercises at 20, 35 and 50% maximal voluntary contraction (MVC) for 60 s. The study was conducted in a climatic chamber with a regulated ambient temperature of 35 degrees C and relative humidity of 50% to induce sweating response at rest by rising skin temperature without a marked change in internal temperature. Sublingual and mean skin temperatures (thermal factors) in both trained and untrained groups were essentially constant throughout all exercise intensities. Changes in heart rate, mean arterial blood pressure, and rating of perceived exertion with increased exercise intensity were similar in both groups. Sweating rate (SR) on the limbs (mean value of forearm and thigh) was significantly greater in the trained group than in the untrained group at 50% MVC (P < 0.05). In addition, the slopes of the relationship between increased SR and exercise intensity (% MVC) on the trunk (chest) and limbs were significantly greater in the trained group than in the untrained group (P < 0.05). Our results suggest that the sweating response caused by non-thermal factors against a background of changing thermal factors was enhanced by physical training. It is also thought that the enhanced sweating response may be especially evident on the limbs than on the trunk, such as improvement of sweating response associated with thermal factors. PMID- 11851595 TI - Effect of induced metabolic alkalosis on sweat composition in men. AB - To determine whether induced metabolic alkalosis affects sweat composition, 10 males cycled for 90 min at 62.5 +/- 1.3% peak oxygen uptake, on two separate occasions. Subjects ingested either empty capsules (placebo) or capsules containing NaHCO3- (0.3 g kg-1 body mass; six equal doses) over a 2-h period, which commenced 3 h prior to exercise. Arterialized-venous blood samples were drawn prior to and after 15, 30, 60 and 90 min of exercise. Sweat was aspirated at the end of exercise from a patch located on the right scapula region. NaHCO3- ingestion elevated blood pH, [HCO3-] and serum [Na+], whereas serum [Cl-] and [K+] were reduced, both at rest and during exercise (P < 0.05). Sweat pH was greater in the NaHCO3- trial (6.24 +/- 0.18 vs. 6.38 +/- 0.18; P < 0.05), whereas sweat [Na+] (49.5 +/- 4.8 vs. 50.2 +/- 4.3 mEq L-1), [Cl-] (37.5 +/- 5.1 vs. 39.3 +/- 4.2 mEq L-1) and [K+] (4.66 +/- 0.19 vs. 4.64 +/- 0.34 mEq L-1) did not differ between trials (P > 0.05). Sweat [HCO3-] (2.49 +/- 0.58 vs. 3.73 +/- 1.10 mEq L-1) and [lactate] (8.92 +/- 0.79 vs. 10.51 +/- 0.32 mmol L-1) tended to be greater after NaHCO3- ingestion, although significance was not reached (P=0.07 and P=0.08, respectively). These data indicate that induced metabolic alkalosis can modify sweat composition, although it is unclear whether the secretory coil, reabsorptive duct, or both are responsible for this alteration. PMID- 11851596 TI - The repeated bout effect and heat shock proteins: intramuscular HSP27 and HSP70 expression following two bouts of eccentric exercise in humans. AB - Exercise-induced damage significantly and predictably alters indirect indicators of muscle damage after one bout of damaging exercise but this response is dampened following a second bout of the same exercise performed 1-6 weeks later. Previously we have described a marked increase in the levels of heat shock proteins (HSPs) HSP27 and HSP70 in human biceps muscle following one bout of high force eccentric exercise. The purpose of the present study was to examine the intramuscular HSP27 and HSP70 response following two identical bouts of exercise [bout 1 (B1) and bout 2 (B2), separated by 4 weeks] relative to indirect indices of muscle damage. Ten human subjects performed 50 high-force eccentric contractions with their non-dominant forearm flexors; muscle damage of the biceps brachii was evaluated 48 h post-exercise with indirect indices [serum creatine kinase (CK) activity, soreness, isometric maximal voluntary contraction (MVC) force and relaxed arm angle] and immunoblotting of high ionic strength muscle biopsy extracts for both HSPs. Not unexpectedly, the indirect indicators of damage changed dramatically and significantly (P < 0.01) after B1 but had a much smaller response after B2. The magnitude of the HSP response was the same after both bouts of exercise, though the control and exercised samples of B2 demonstrated a lower basal HSP expression. Thus, though both indirect and cellular indicators of exercise-induced muscle damage demonstrate an adaptation consequent to the first bout of exercise, these adaptations are quite different. It is possible that the lower basal HSP expression of the cellular response mediates the attenuation of damage associated with B2 as indicated by indirect indices. PMID- 11851597 TI - Differential response of muscle phosphocreatine to creatine supplementation in young and old subjects. AB - This study compared the effects of short-term creatine supplementation on muscle phosphocreatine, blood and urine creatine levels, and urine creatinine levels in elderly and young subjects. Eight young (24 +/- 1.4 years) and seven old (70 +/- 2.9 years) men ingested creatine (20 g day-1) for 5 days. Baseline muscle phosphocreatine measurements were taken pre- and post-supplementation using nuclear magnetic resonance spectroscopy (NMR). On the first day of supplementation subjects had blood samples taken immediately before and hourly for 5 h following ingestion of 5 g of creatine, and a pharmacokinetic analysis of plasma creatine levels was conducted. Twenty-four hour urine collections were conducted for 2 days prior to the supplementation period and for 5 days during supplementation. Old subjects had significantly higher baseline plasma creatine levels than young subjects (68.5 +/- 12.5 vs. 34.9 +/- 4.7 micromol L-1; P < 0.02). There were no significant differences between groups in plasma creatine pharmacokinetic parameters (i.e. area under the curve, elimination rate constant, absorption rate constant, time to maximum concentration, and maximum concentration) following the 5 g oral creatine bolus. Urine creatine, assessed pre and on 5 days of supplementation, increased (P < 0.001), with no difference between groups. Urine creatinine did not change as a result of creatine supplementation. Young subjects showed a significantly greater increase in muscle phosphocreatine compared with old subjects, and post-supplementation muscle phosphocreatine levels were greater in young subjects (young 27.6 +/- 0.5; old 25.7 +/- 0.8 mmol kg-1 ww) (P=0.02). There were no differences in blood or urine creatine between groups in response to supplementation, but old subjects had a relatively small increase (young 35% vs. old 7%) in muscle phosphocreatine after supplementation. PMID- 11851598 TI - Biphasic effects of substance P on respiratory activity and respiration-related neurones in ventrolateral medulla in the neonatal rat brainstem in vitro. AB - The effects of substance P (SP) on respiratory activity in the brainstem-spinal cord preparation from neonatal rats (0-4 days old) were investigated. The respiratory activity was recorded from C4 ventral roots and intracellularly from three types of respiration-related neurones, i.e. pre-inspiratory (or biphasic E), three subtypes of inspiratory; expiratory and tonic neurones in the ventrolateral medulla (VLM). After the onset of SP bath application (10 nM-1 microM) a dose-dependent decline of burst rate (by 48%) occurred, followed by a weaker dose-dependent increase (by 17.5%) in burst rate. The biphasic effect of SP on inspiratory burst rate was associated with sustained membrane depolarization (in a range of 0.5-13 mV) of respiration-related and tonic neurones. There were no significant changes in membrane resistance in any type of neurones when SP was applied alone or when synaptic transmission was blocked with tetrodotoxin (TTX). The initial depolarization was associated with an increase in inspiratory drive potential (by 25%) as well as in bursting time (by 65%) and membrane excitability in inspiratory and pre-inspiratory neurones, which corresponded to the decrease in burst rate (C4 activity). The spiking frequency of expiratory and tonic neurones was also increased (by 36 and 48%). This activation was followed by restoration of the synaptic drive potential and bursting time in inspiratory and to a less extent in pre-inspiratory neurones, which corresponded to the increase in burst rate. The discharge frequency of expiratory and tonic neurones also decreased to control values. This phase followed the peak membrane depolarization. At the peak depolarization, SP reduced the amplitude of the action potential by 4-8% in all types of neurones. Our results suggest that SP exerts a general excitatory effect on respiration-related neurones and synaptic coupling within the respiratory network in the VLM. The transient changes in neuronal activity in the VLM may underlie the biphasic effect of SP in the brainstem respiration activity recorded in C4 roots. However, the biphasic effect of SP on inspiratory burst rate seems to be also defined by the balance in activity of other SP-sensitive systems and neurones in the respiratory network in the brainstem and spinal cord, which can modify the activity of medullary respiratory rhythm generator. PMID- 11851600 TI - Benign lateralizing haematuria: the impact of upper tract endoscopy. PMID- 11851601 TI - Retrograde ureteropyeloscopic holmium laser lithotripsy for large renal calculi. AB - OBJECTIVE: To assess the efficacy and safety of the retrograde ureteropyeloscopic holmium laser for treating renal stones that are too large to treat with extracorporeal shock wave lithotripsy (ESWL). PATIENTS AND METHODS: Thirty patients (22 men and eight women, mean age 43 years, range 18-62) with a renal stone burden of > 2 cm were selected for laser treatment. The stones were in the renal pelvis in 16 patients, lower calyx in five, middle calyx in two, upper calyx in one and multiple pelvic and calyceal in six. Lithotripsy was undertaken using a holmium laser through 550 microm and 200 microm laser fibres passed through a semi-rigid fibre-optic long ureteroscope or the actively deflectable flexible ureteropyeloscope, respectively. Success was defined as total fragmentation of the stone to < 2 mm in diameter and/or clear imaging on renal ultrasonography and plain films within the 3-month follow-up. Patients in whom the treatment failed received either alternative therapy or complementary ESWL. RESULTS: Endoscopic access and complete stone fragmentation was achieved in 23 of the 30 patients (77%). The treatment failed in seven patients because of poor visualization, the initial presence of stones in, or migration of their large fragments to, an inaccessible calyx. There were no major intraoperative complications. Minor complications after treatment included haematuria that persisted for 2 days in one patient and high-grade fever in two patients; all were treated conservatively. CONCLUSION: Large renal calculi that are not amenable to ESWL monotherapy can be safely and effectively treated with a retrograde endoscopic technique that seems to compete well with the more invasive percutaneous or open surgical manoeuvres. PMID- 11851602 TI - Pelvicalyceal stone load: a factor affecting the outcome of extracorporeal shockwave lithotripsy for renal pelvic calculi. AB - OBJECTIVES: To determine the effect of variations in pelvicalyceal volume on the results of extracorporeal shock wave lithotripsy (ESWL) and the relationship of stone load to estimated pelvicalyceal volume in patients with renal pelvic stones. PATIENTS AND METHODS: In all, 204 patients with renal pelvic stones were treated primarily by ESWL. Exclusion criteria were radiolucent stones, dilatation of the renal collecting system, JJ stent insertion before ESWL and an inadequate follow-up. The surface area of the stones and the renal collecting system were measured using a grid of 1 mm2 divisions. The pelvicalyceal volume was calculated as 0.6 (area)1.27. To evaluate the treatment results a pelvicalyceal stone load (PSL) index, describing the relationship of stone load to total estimated pelvicalyceal volume, was defined as stone volume/renal collecting system volume. RESULTS: The mean (range) pelvicalyceal volume of the patients was 13.24 (4.12 28.47) mm3 and the mean PSL index was 26.2%. The cumulative success rates according to the PSL decreased from 97% to 90% with increasing PSL, but remained at > 95% in patients with a PSL index of < 50%. In all, 184 sessions were applied and the session/patient ratio increased from 1.41 to 3.0 with increasing PSL. Complications were acute pyelonephritis in three and steinstrasse in five patients. CONCLUSION: The PSL index appears to be a more accurate and reproducible method for predicting the outcome of ESWL, and has the advantage of considering the effect of pelvicalyceal anatomy. PMID- 11851603 TI - (L)-2-oxothiazolidine-4-carboxylate in the treatment of primary hyperoxaluria type 1. AB - OBJECTIVE: To evaluate the short-term efficacy of (l)-2-oxothiaolidine-4 carboxylate (OTZ, which reduces urinary oxalate excretion in normal subjects) in the treatment of primary hyperoxaluria type 1 (PH1) in a phase II study. PATIENTS AND METHODS: Two patients with PH1 received intravenous infusions of OTZ (100 mg/kg body weight for 2 h) given every 8 h for four doses. One patient also received a placebo treatment. Urine samples (24-h collections) were obtained before and during OTZ treatment and assayed for oxalate, citrate, creatinine, sulphate and pH. Daily blood samples were assayed for plasma oxalate and serum creatinine. RESULTS: Urinary oxalate excretion was unaffected by OTZ treatment. Plasma oxalate declined in both individuals with OTZ treatment, but the effect was small. Plasma cysteine was normal in one patient, rising from a mean (sd) of 36 (3.7) micromol/L before treatment to a peak of 141 micromol/L after OTZ, but was not detected in samples from the other patient. The ratio of oxalate to creatinine clearances was high in both patients, with mean values of 3.1 and 3.8. CONCLUSIONS: Treatment with OTZ did not lead to clinically significant changes in urinary oxalate excretion. The high clearance of oxalate in these patients suggests a substantial renal secretion of oxalate. PMID- 11851604 TI - Facilitating a conceptual shift: psychological consequences of interstitial cystitis. PMID- 11851605 TI - A study to detect Gardnerella vaginalis DNA in interstitial cystitis. AB - OBJECTIVE: To investigate the possible role of Gardnerella vaginalis in interstitial cystitis (IC), using molecular methods to avoid difficulties with the culture and recovery of viable organisms, and the problems associated with the recovery of low numbers of culturable organisms. MATERIALS AND METHODS: Thirty-three bladder biopsy samples (29 paraffin-embedded and four freshly frozen) from patients with IC, diagnosed according to National Institute of Diabetes, Digestive and Kidney Diseases criteria, were assessed. Biopsy samples were used as urine samples may be contaminated by normal vaginal flora. A positive control comprised a 'normal' biopsy sample from a patient with a previous bladder tumour, seeded with G. vaginalis NCTC 10915. Microbial DNA was extracted from all paraffin-embedded and fresh specimens, and subjected to in vitro amplification by polymerase chain reaction (PCR) with G. vaginalis-specific primers. RESULTS: The anticipated PCR product of 333 base pairs was obtained with the positive control, whereas none of the other biopsy samples showed positive amplification specific for G. vaginalis. CONCLUSION: As there was no G. vaginalis DNA in any of the samples from patients with IC, it is an unlikely candidate in the pathogenesis of IC. PMID- 11851606 TI - A systematic review of tension-free urethropexy for stress urinary incontinence: intravaginal slingplasty and the tension-free vaginal tape procedures. PMID- 11851607 TI - Urinary stress incontinence in obese women: tension-free vaginal tape is the answer. AB - OBJECTIVE: To determine the effectiveness of the tension-free vaginal tape (TVT) in obese women with genuine stress incontinence (GSI), in whom obesity is often considered a relative contraindication to surgical treatment by traditional approaches, e.g. Burch colposuspension and slings (which are difficult and carry increased morbidity) or injectable agents (which although simple, are unpredictable and expensive). PATIENTS AND METHODS: Data on 242 consecutive women with urodynamically proven GSI were collected prospectively. The women were subdivided into three groups with a body mass index (BMI) of < 25, 25-29 and >or=30; obesity was defined as a BMI of >or=30. All procedures were performed under spinal anaesthesia. The King's validated quality of life (QoL) questionnaires (version 7) were completed before and 6 months after surgery. The subjective results were defined as a cure, significant improvement or failure. RESULTS: Almost 90% of the obese women with GSI were cured, while the remaining 10% noted a considerable improvement in their symptoms. There was no significant difference in cure rates among the three groups. There was a highly significant (P < 0.001) improvement in QoL in all groups. CONCLUSION: The TVT is at least as effective in obese women as in those with a lower BMI. The TVT is a simple and minimally invasive procedure, with low morbidity even in the obese group. TVT can be offered confidently to all obese women with GSI. PMID- 11851608 TI - The surgical results of the pubovaginal sling procedure using polypropylene mesh for stress urinary incontinence. AB - OBJECTIVE: To investigate the surgical results after a pubovaginal sling procedure using polypropylene mesh in women with stress urinary incontinence (SUI). PATIENTS AND METHODS: Sixty-four women with different types of SUI underwent a pubovaginal sling procedure, using polypropylene mesh. The sling was placed at the level of the proximal half of the urethra and tied with adequate tension, but not obstructing the bladder outlet. A video-urodynamic study and transrectal ultrasonography were undertaken before and after surgery. The surgical results were assessed and the urodynamic changes compared at different stages. RESULTS: At a median follow-up of 24 months, 52 patients (81%) were completely continent, 10 (16%) had an improvement but with mild SUI, and two had persistent SUI requiring a second sling procedure. The treatment result was considered satisfactory by 55 patients (86%). The video-urodynamic study showed no significant change in voiding pressure, cystometric capacity and residual urine volume after surgery. The bladder neck opening time was increased at 7 days and the maximum flow rate increased at 3 months after surgery. Transrectal ultrasonography showed that all the polypropylene mesh slings were located beneath the bladder neck and proximal urethra, with no notable granuloma formation around the sling. CONCLUSION: The pubovaginal sling procedure is effective in treating female SUI, using polypropylene mesh as the sling material. The video-urodynamic results showed that a pubovaginal sling of polypropylene mesh causes no bladder outlet obstruction when the correct surgical technique is used. PMID- 11851609 TI - A new intravaginal device for stress incontinence in women. AB - OBJECTIVE: To compare two versions of the same type of disposable intravaginal device (the Conveen Continence Guard, CCG, and the Contrelle Continence Tampon, CCT, Coloplast a/s, Humlebaek, Denmark) for treating stress incontinence in women. PATIENTS AND METHODS: Women with the predominant symptom of stress incontinence were recruited from four centres in Denmark, Australia and the UK. The women were assessed using a 24-h pad-test, uroflowmetry, postvoid residual urine volume and a voiding diary before treatment, and after 5 weeks using each of the two devices. Vaginal swabs and specimens of urine were sent for culture, and a questionnaire about the subjective effect and adverse events completed at each visit. In all, 94 women were recruited, of whom 62 (66%) completed the study. RESULTS: Both devices reduced the amount of leakage significantly, but the CCT reduced urine loss significantly more than the CCG. Uroflowmetry values and residual urine volume were unchanged when using the two devices. Vaginal culture showed no abnormality during the study period, and only one woman was treated for a urinary tract infection. Side-effects were few and not serious. The women found both devices easy to prepare, insert and use; two-thirds preferred the CCT to the CCG. CONCLUSION: The new intravaginal device (CCT) is more effective for treating stress incontinence than the currently available version (CCG), and patient acceptability of the new device seems to be superior. PMID- 11851610 TI - Incontinence aids in Sweden: users and costs. AB - OBJECTIVE: To study age- and sex-specific use and costs of incontinence aids distributed free of charge in Sweden. SUBJECTS AND METHODS: The study was conducted in the county of Jamtland, Sweden (132,000 inhabitants). The use and cost of incontinence aids for people living in their homes and the total cost of incontinence aids for residents of special accommodation (e.g. nursing homes, homes for the elderly and sheltered housing) was obtained from a central database constructed for the purpose. Individual usage of incontinence aids by those in special accommodation was studied in two districts of Jamtland, representing 18% of the population. RESULTS: Free incontinence products were used by 6.4% of all women and 2.4% of all men in the county. There was a sharp increase in usage from the age of 75 years. Of the users, 21% lived in special accommodation. If the data from Jamtland are extrapolated nationally, then 274,000 women and 93,000 men in Sweden (total population 8.8 million) are using free incontinence products. The total cost of incontinence aids for Jamtland during 1999 was 15.4 million Swedish krona (SK), and those in special accommodation accounted for 46% of these costs. This corresponds to an estimated total cost in Sweden of approximately 925 million SK. Although 75% of the users were women, women only contributed 61% of the total costs. The mean annual cost of incontinence aids for an incontinent man was twice that of an incontinent woman. More than half of the costs were attributable to those aged >or=80 years. CONCLUSIONS: The estimated national costs of free incontinence aids accounts for 0.5% of the total costs of Swedish healthcare, including the care and nursing of older and disabled people, and for 0.05% of the gross national product. PMID- 11851611 TI - Transcutaneous electrical nerve stimulation in the treatment of neurological patients with urinary symptoms. AB - OBJECTIVE: To determine whether transcutaneous electrical nerve stimulation (TENS) benefits patients with urinary symptoms caused by neurological diseases. PATIENTS AND METHODS: Patients with urinary symptoms from any kind of neurological disease were prospectively recruited between October 1996 and July 1998. Before attending the first assessment patients were asked to complete a week's diary recording the frequency of micturition, incontinence episodes, and frequency of pad and clothes changing. At the first assessment the patients completed the Frimodt-Moller urinary symptom questionnaire, and quality-of-life scales including the Nottingham Health Profile and Short-Form 36. Demographic and disability data (Barthel Index and Frenchay Aphasia Screening Test) were recorded, and patients underwent a neurological examination and urodynamic studies. The placing of electrode pads on the sacral dermatomes 2.5 cm either side of and 2.5 cm above the natal cleft was demonstrated, and the patient instructed to use TENS for 90 min twice a day. The current strength applied was set to that which the patient could tolerate, at a square-wave of 20 Hz and 200 micros duration. Six weeks later the patients were further assessed, where the diary exercise, questionnaires and urodynamics were repeated. In all, 44 patients (13 men and 33 women, mean age 50.8 years) were recruited. RESULTS: The commonest disease was multiple sclerosis and the commonest impairments para/tetraplegia or hemiplegia. There was no change in the neurological status of the 34 patients completing the study. Irritative voiding symptoms were significantly decreased (0.68-0.61, P = 0.003) and diaries also showed significant improvements in the 24 h frequency of micturition (P = 0.01), incontinence episodes (P = 0.04) and clothes changes (P = 0.02). Urodynamics showed detrusor hyper-reflexia in most patients. The only significant changes after TENS were an increased postvoid residual volume (from a mean of 134 mL to 160 mL, P = 0.03) and an increase in the volume leaked during the urodynamic study with TENS on (from a mean of 4.7 mL to 12 mL, P = 0.003). There were no significant changes in the quality-of-life scores. Of the 34 patients completing the study, half still reported a benefit from TENS at 1 year, although some patients found it took 3-4 weeks to work. CONCLUSION: TENS applied to the sacral dermatomes of neurological patients with urinary symptoms had a minimal effect on urodynamic data but significantly improved irritative urinary symptoms, 24-h urinary frequency, incontinence and clothes changing. The lack of effect on quality-of-life measures probably reflects the lack of sensitivity in the tools used in this group of patients. We therefore recommend using TENS in this often problematical group of patients. PMID- 11851612 TI - The role of volume-weighted mean nuclear volume in predicting tumour biology and clinical behaviour in patients with prostate cancer undergoing watchful waiting. AB - OBJECTIVE: To investigate whether the volume-weighted mean nuclear volume (MNV, the only means by which unbiased estimates of three-dimensional variables can be obtained from a two-dimensional section by stereological methods) at diagnosis correlates with tumour biology and clinical behaviour in patients with prostate cancer treated by watchful waiting. PATIENTS AND METHODS: In a prognostic study, 64 patients with clinically localized prostate cancer were followed prospectively with initial expectant management. The median (mean, range) follow-up was 22 (27, 6.0-68) months. The prostate specific antigen (PSA) doubling time (PSADT) was calculated by linear regression. The MNV was estimated using biopsy specimens, based on a stereological method, and compared with PSADT and traditional clinicopathological variables. RESULTS: PSADT was significantly associated with MNV, but not with other clinicopathological variables. The PSA 'rapid-riser' subset (PSADTor=median value) and PSA-stable subsets (P = 0.0017 and 0.004, respectively). On multivariate analysis using a stepwise Cox proportional hazards regression, only MNV remained independently significant as a predictor of clinical progression among the clinicopathological variables (P < 0.001). CONCLUSIONS: These findings suggest that cancer cell nuclear volume is significantly associated with tumour biology and behaviour in patients with prostate cancer. Although further study with a larger patient population is needed to confirm the findings, estimates of MNV may be an important prognostic indicator in men treated with watchful waiting. PMID- 11851613 TI - Renal transplantation: the impact of schistosomiasis. PMID- 11851614 TI - Cyclooxygenase-2: a possible target in schistosoma-associated bladder cancer. AB - OBJECTIVE: To analyse and compare the expression of cyclooxygenase (COX) enzymes in schistosoma-associated bladder cancer, and to determine any association with tumour grade or stage. MATERIALS AND METHODS: Sixty paired samples of tumour and adjacent nonmalignant urothelium were identified. There were 25 squamous and 28 transitional cell carcinomas, and seven adenocarcinomas. Serial sections were obtained and a standard three-layer immunohistochemistry protocol, using COX-1- and COX-2-specific mouse monoclonal antibodies, applied. RESULTS: COX-1 was expressed mostly in nonvascular smooth muscle with weak reactivity in malignant and nonmalignant urothelium. Nonmalignant urothelium expressed COX-2 weakly, notably in areas of dysplasia and squamous metaplasia whereas there was a significant increase in COX-2 (P < 0.001) with moderate to strong granular cytoplasmic expression in all three malignant histological types. The COX-2 reactivity was higher in transitional and adenocarcinomas than in squamous cell carcinoma (P < 0.001). Areas of carcinoma in situ showed COX-2 reactivity comparable with that in invasive areas and more intense than that detected in dysplastic or metaplastic urothelium (P < 0.001). There was a statistically significant positive correlation between COX-2 expression and tumour grade (P = 0.0052). CONCLUSION: COX-2 is over-expressed in schistosoma-associated bladder cancer, consistent with a potential role for COX-2 inhibitors in the prevention and management of this disease. PMID- 11851615 TI - Open stone surgery in children: is it justified in the era of minimally invasive therapies? AB - OBJECTIVE: To review experience with open surgery for paediatric urolithiasis during a 10-year period. PATIENTS AND METHODS: The hospital records of patients up to 13 years old and treated between 1990 and 2000 for stones were reviewed; there were 310 patients (98 girls and 212 boys, aged 9 months to 13 years, mean 6.8 years). RESULTS: The commonest symptoms were renal-ureteric colic (26.7%), gross haematuria (19%), urinary retention (16.7%), and abdominal and/or flank pain (13.2%). Because of poverty and the resultant inability to pay medical fees, 19 children presented very late with pyonephrosis (resembling peritonitis in nine) and obstructive renal atrophy in 23. In 18 other patients the delay was caused by the disappearance of pain. Delayed presentation was the most important factor in developing complications from the stone. The stones were in the calyces in 15 patients, the pelvis in 113, the ureter in 56, the bladder in 71, the urethra in 17 and in a combination of sites in 38. The mean (range, median) stone size was 27 (9-75, 22) mm; 80 (25.8%) were complete staghorn stones. Indications for open surgery were a complex stone burden (62%), ESWL failure (14.5%), need for nephrectomy (1.9%), anatomical abnormalities (2.2%), and unavailability of minimally invasive alternatives (19%). All of the nephrectomized patients underwent unilateral stone removal and contralateral nephrectomy. For parents, the cost and reliability of the result were more important than other considerations, e.g. having a large or small incision. The overall stone-free rate at discharge was 95.4% (100% for single stones). In five cases (1.6%) a repeat open procedure was needed. The mean (range, median) hospital stay was 4 (1 13, 3) days. CONCLUSIONS: Arguments against open surgery for urolithiasis in adults should not be extrapolated to children, in whom open surgery is safe and effective. In Iran and many 'developing' countries, open surgery is less expensive, more effective, more dependable, and more easily available than minimally invasive alternatives. At least in such countries it deserves to be among the first-line therapies for paediatric urolithiasis, and urologists in less-developed countries should not decline to offer open surgery because it is almost obsolete in developed countries. Delayed presentation (through poverty and/or unawareness) contributes significantly to the morbidity of urolithiasis. PMID- 11851616 TI - Decreased nocturnal urinary antidiuretic hormone excretion in enuresis is increased by imipramine. AB - OBJECTIVE: To assess the role of integrated nocturnal antidiuretic hormone (ADH) secretion in children with enuresis, and possible modifications induced by treatment with imipramine. PATIENTS AND METHODS: The morning plasma ADH and nocturnal urinary ADH integrated concentrations were measured in 18 consecutive enuretic children (patients) and 21 age- and sex-matched controls admitted for minor treatment. Diurnal and nocturnal urine production, and plasma and urinary osmolality were also determined; lumbosacral radiography and uroflowmetry were undertaken in the patients. The assessments were repeated after 14 days of treatment with imipramine hydrochloride (orally, 20 mg/night). RESULTS: Half the patients had occult spinal malformations but the uroflowmetry results were all within the normal range. The median (95% confidence interval, CI) urinary ADH integrated concentrations were markedly lower in patients, at 29.7 (22.1-37.3) vs 63.0 (35.1-90.8) pg/mL/h (P = 0.03) than in controls. Plasma ADH levels were significantly increased by imipramine (0.64 to 1.47 pg/mL, 95% CI, 0.40-0.89 vs 0.26-3.2; P < 0.001), as were nocturnal urinary ADH integrated concentrations, at 29.7 (22.1-37.3) vs 59.0 (37.3-80.6) pg/mL/h (P < 0.001), and morning plasma osmolality decreased, from 298.5 (294.5-302.5) to 294.9 (292.4-297.3) mosmol/kg (P = 0.003), as was the 24-h fluid intake, from 983 (721-1245) to 666 (435-897) mL (P = 0.004). CONCLUSIONS: We conclude that enuretic children have a lower nocturnal ADH excretion; imipramine restores nocturnal ADH excretion, increases morning plasma ADH levels, and causes consistent changes in other biochemical variables. PMID- 11851617 TI - Increased morbidity after circumcision from a hidden complication. AB - OBJECTIVE: To describe a late complication of circumcision and to elucidate the role of surgical materials or any foreign body in the development of a subcutaneous mass in circumcised boys. PATIENTS AND METHODS: From May 1998 to March 2001, 646 boys were circumcised and 523 (mean age 6.5 years, range 0-13) re examined for the possible development of a subcutaneous mass. Twenty-six (5%) of the re-examined patients had such a mass under the penile skin; it was removed in all patients under local anaesthesia and examined histopathologically. RESULTS: The mean (sd, range) delay after circumcision before developing or detecting the mass was 3.2 (0.7, 1-7) months. All patients were asymptomatic but there was purulent discharge in four. On removing the mass, histopathology showed the development of granulation tissue with foreign-body giant cells. CONCLUSION: There may be minor complications after circumcision which cannot be avoided even when the procedure is undertaken by surgeons. Awareness of such complications occurring long after circumcision may aid in the early detection of this asymptomatic mass and prevent a more severe outcome. PMID- 11851618 TI - Prevalence and characteristics of cryptorchidism in a Nigerian district. AB - OBJECTIVE: To determine the prevalence and characteristics of cryptorchidism among primary schoolboys in a Nigerian district. SUBJECTS AND METHODS: The district selected had 35 primary schools with 23,342 pupils, consisting of 11,275 girls and 11,967 boys. Using a cluster-sampling technique, five primary schools were visited; 1096 boys (aged 5-13 years) participated in the study, giving a sampling ratio of 1:11. The boys underwent a clinical examination of the groin, scrotum and abdomen. RESULTS: Cryptorchidism was found in nine subjects, giving a prevalence rate of 0.82%. All the undescended testes were unilateral, of which five were right-sided. Eight of the testes were intracanalicular and one was at the external ring. There were no cases of orchidopexy. CONCLUSION: The prevalence of cryptorchidism among primary schoolboys in this district of Nigeria was high, at eight per 1000. Delayed diagnosis and treatment remains a problem because of the prevailing socio-economic factors. However, the incidence of cryptorchidism was similar to that reported in other parts of the world. PMID- 11851619 TI - Prostatic fluid-free insulin-like growth factor-1 in relation to prostate cancer. AB - OBJECTIVES: To establish the feasibility of measuring free insulin-like growth factor-1 (IGF-1) in prostatic fluid and to determine if there is a difference in free IGF-1 levels in the prostatic fluid of patients with prostatic adenocarcinoma and in normal men. PATIENTS AND METHODS: Prostatic fluid samples were collected from men with histologically confirmed prostate cancer and from control subjects with no malignancy. Prostatic fluid and serum free IGF-1 levels were measured in duplicate by radioimmunoassay, and the differences analysed using the Mann-Whitney U-test and Pearson correlation. RESULTS: Prostatic fluid free IGF-1 concentrations did not differ significantly (P = 0.23) and there was no difference in serum free IGF-1 levels between the groups. While serum PSA level was significantly correlated with age (P < 0.001) the free IGF-1 level of prostatic fluid was not. There was also no correlation between prostatic fluid free IGF-1 and serum PSA levels. CONCLUSIONS: Free IGF-1 can be detected in prostatic fluid with acceptable sensitivity. However, the free IGF-1 level in prostatic fluid does not help to detect patients with prostate cancer and is not a tumour marker. PMID- 11851620 TI - Developmental changes in the biochemical and functional properties of endothelin receptors in rabbit renal pelvis. AB - OBJECTIVE: To examine the effect of age on the biochemical and functional properties, and regional distribution of endothelin (ET) receptors in the rabbit renal pelvis. MATERIALS AND METHODS: The properties of ET receptors in 6-week-old and 6-month-old male rabbit renal pelves were examined using isolated muscle-bath and radioligand receptor-binding techniques. RESULTS: ET-1 caused a significant increase in the contractile force in muscle strips from all regions of the renal pelvis from both age groups, with the following rank order: upper=middle>lower. The magnitude of the ET-1-induced contractile responses were similar in the lower pelvic regions in both ages, but the responses in the upper and middle regions were significantly greater in younger rabbits. ET-1 increased the frequency of spontaneous activity in a concentration-dependent manner in the upper and middle pelvic regions in both age groups, with significantly smaller ED50 values in the younger than in the older rabbits. In both age groups the lower pelvic region lacked spontaneous activity. The density of total ET receptors was higher in the upper and middle regions of the renal pelvis than in the lower renal pelvis of both ages, with the density in the upper and middle regions being greater in older than in younger rabbits. In all regions, ET subtype selective compounds inhibited [125I]ET-1, binding consistent with the predominance of the ETA receptor subtype, except in the lower region of the older rabbits, in which the densities of ETA and ETB subtypes were similar. In all regions, the younger renal pelvis contained a higher proportion of ETA receptors than in older tissues. Light microscopic autoradiographic data indicated the presence of ETA and ETB receptors in smooth muscle and epithelial cells, respectively. CONCLUSION: These data indicate the presence of regional differences in the density of ET receptors and in the contractile responses to ET-1 in rabbit renal pelvis, and that although older rabbit renal pelvis contains more total ET receptors than younger renal pelvis, the latter had a higher portion of the ETA receptor subtype and the younger tissues were more responsive to ET-1. PMID- 11851621 TI - Higher expression of K-ras is associated with parathyroid hormone-related protein induced hypercalcaemia in renal cell carcinoma. AB - OBJECTIVES: To determine whether the K-ras oncogene is associated with parathyroid hormone-related protein (PTHrP) production in renal cell carcinoma (RCC) and whether the serum value of PTHrP is related to the patients' survival. PATIENTS AND METHODS: The serum levels of PTHrP and corrected serum calcium levels were analysed in 51 consecutive patients (29 men and 22 women, mean age 63.7 years, range 33-82) with newly diagnosed RCC. Matched pairs were analysed of the mRNA levels of K-ras and PTHrP in tumour and in corresponding non-tumour tissue originating from the same patient, using the polymerase chain reaction after reverse transcription. RESULTS: Seven patients had elevated serum PTHrP values at the diagnosis of RCC. The mRNA expression of K-ras and PTHrP were detected in both tumour and non-tumour tissues, with K-ras mRNA levels being higher in the former than the latter (P < 0.05), and correlated with tumour stage (P < 0.05). There were no differences in PTHrP mRNA levels between the tissues. Furthermore, the mRNA levels of K-ras and PTHrP in seven tumours from patients with high serum values of PTHrP were higher than in tumours from those with normal values (both P < 0.01). The expression of mRNAs of K-ras and PTHrP was positively correlated (r = 0.771, P < 0.001). In seven patients with high serum PTHrP values the mRNA levels of PTHrP correlated with serum values of PTHrP and calcium (r = 0.875, P < 0.01 and r = 0.762, P < 0.05, respectively). Kaplan-Meier plots of survival rate in patients with elevated or normal serum PTHrP showed that high serum PTHrP was associated with a shorter overall survival (P < 0.05). The Cox proportional hazards model showed that serum PTHrP was an independent predictor of overall survival (P < 0.05). CONCLUSIONS: These findings suggest that K-ras may be associated with PTHrP-induced hypercalcaemia and that PTHrP levels may reflect the aggressiveness of tumour cells through the K-ras oncogene in RCC. PMID- 11851622 TI - Nitric oxide synthase patterns in normal and varicocele testis in adolescents. AB - OBJECTIVE: To determine if the testes of normal adolescents can produce nitric oxide (NO), by assessing NO synthase (NOS) activity, and whether this activity changes in adolescents with left idiopathic varicocele. PATIENTS AND METHODS: After obtaining informed consent, testicular biopsies were obtained from eight adolescents (mean age 16.4 years; controls) who underwent surgery for inguinal hernia or hydrocele, and from 20 adolescents (mean age 16.2 years) operated for left idiopathic varicocele. Inducible and endothelial NOS (iNOS and eNOS) isoforms were investigated in the biopsy specimens by immunohistochemical localization and Western blot analysis using specific fluorescein-conjugated antibodies. RESULTS: Both normal and pathological samples expressed eNOS at the level of vessels and Leydig cells. The iNOS was expressed in Leydig cells of normal testes and over-expressed in Leydig cells of varicocele testes. CONCLUSION: Leydig cells of adolescent testes constitutively express iNOS. Under pathological conditions, e.g. varicocele, iNOS is up-regulated and is a possible source of NO overproduction. These results could be useful in explaining the pathogenesis of both testis and sperm dysfunction in varicocele. PMID- 11851623 TI - Relationship between serum ferritin levels and tumour status in patients with renal cell carcinoma. AB - OBJECTIVE: To assess the relationship between serum ferritin levels (a useful marker for diagnosing and staging renal cell carcinoma, RCC) and tumour status in RCC of =0.1 IU/mL). The sensitivity, specificity and inter-rater agreement (K or Kw) of each method were related to the in vitro NT selected as the reference method. RESULTS: The in vivo NT test corresponded very well with the in vitro NT in its ability to differentiate between protection/relative protection and no protection (sensitivity 97%, specificity 87% and K=0.84). The EIA test showed a high sensitivity (96%), but since many sera were categorized as protected rather than not protected, the specificity (30%) and inter-rater agreement (K=0.29) were low. The PHA test had a very high specificity (100%) but a low sensitivity (86%). CONCLUSIONS: The agreement between the two neutralization tests was high. If none of the neutralization assays is routinely available, the PHA test can be used to predict the need for vaccination on an individual basis but should not be used for seroepidemiologic studies, since the protection rate for diphtheria would be falsely too low, due to the lower sensitivity. The indirect EIA test used in this study should not be used routinely. PMID- 11851695 TI - Failure of bacteriophage typing to detect an inter-hospital outbreak of methicillin-resistant Staphylococcus aureus (MRSA) in Zagreb subsequently identified by random amplification of polymorphic DNA (RAPD) and pulsed-field gel electrophoresis (PFGE). AB - OBJECTIVE: To establish the extent of inter-hospital spread of methicillin resistant Staphylococcus aureus (MRSA) in Zagreb and to determine the most suitable method for typing local strains. METHODS: We analyzed a collection of 33 MRSA isolates from three Zagreb hospitals together with five unrelated British MRSA isolates by antibiogram typing, bacteriophage typing, randomly amplified polymorphic DNA (RAPD) analysis and pulsed-field gel electrophoresis (PFGE) after digestion with Smal restriction endonuclease. Bacteriophage typing was done with the international set of S. aureus typing phages. RAPD and PFGE profiles were analyzed visually and by using the 'GelCompar' computer program. RESULTS: Antibiogram typing provided eight profiles. Thirty (91%) of the 33 Croatian strains of MRSA were non-typable by phage typing. Visual analysis of RAPD products identified six, and visual analysis of PFGE fragments nine, distinct profiles. Computer analysis of RAPD data separated British isolates from the Croatian ones, but did not cluster the visually determined RAPD types. PFGE computer analysis separated British isolates and clustered isolates in concordance with visual interpretation. Thirty-one of the 38 isolates (82%) were visually grouped in the same clusters by both molecular methods. The dominant strain was present in each of the three hospitals. CONCLUSIONS: Bacteriophage typing was unhelpful for the analysis of Croatian MRSA, since most strains were untypable with the international set of bacteriophages. RAPD and PFGE were more successful in typing the organisms and showed evidence of inter-hospital spread of one predominant MRSA strain in all three Zagreb hospitals. Thus RAPD and PFGE proved to be a useful aid in elucidating the epidemiology of MRSA infection in Zagreb hospitals and should be established in Croatia for typing MRSA. PMID- 11851696 TI - Vertebral osteomyelitis caused by Mycobacterium avium-intracellulare in a patient with AIDS. PMID- 11851697 TI - Shiga toxin-producing Escherichia coli was the third most frequent bacterial cause of diarrhea in Austria during July and August of 1998. PMID- 11851698 TI - Neisseria cinerea, a bacterium whose bacteriological identification is difficult. PMID- 11851699 TI - Extended-spectrum beta-lactamase TEM-4 in Pseudomonas aeruginosa. PMID- 11851700 TI - Disseminated tuberculosis and idiopathic CD4+ T-lymphocytopenia. PMID- 11851702 TI - 10th European Congress of Clinical Microbiology and Infectious Diseases (ECCMID), Stockholm, Sweden 28--31 May 2000. PMID- 11851701 TI - Do the quinolones still constitute valid empirical therapy for community-acquired urinary tract infections in Spain? PMID- 11851703 TI - Review on evaluations of currently available blood-culture systems. PMID- 11851704 TI - Role of nasopharyngeal bacterial flora in the evaluation of recurrent middle ear infections in children. AB - OBJECTIVE: To study the nasopharyngeal colonization in otitis-prone children before and after adenoidectomy. METHODS: The study population consisted of 35 children between 11 months and 4 years of age, undergoing adenoidectomy and tube placement for recurrent acute otitis media. All these children were otitis prone (OP). During general anesthesia, bacteriologic samples were obtained from the nasopharynx and the middle ear fluid, if present. During the follow-up visit, a new nasopharyngeal culture was taken. The control population consisted of 35 children undergoing surgery for non-ear-nose-throat pathology. These children had no history of recurrent upper respiratory tract infections. RESULTS: Colonization of the nasopharynx with potential pathogens (Streptococcus pneumoniae, Moraxella catarrhalis, Haemophilus influenzae) occurred significantly more in the OP children than in the non-OP children. Adenoidectomy resulted in a substantial decrease of potential middle ear pathogens in the nasopharynx and an increase of normal commensal flora. In about half of the patients, middle ear fluid was still present at the time of tube placement; in most instances, H. influenzae was cultured. Typing with arbitrarily primed PCR indicated substantial genetic diversity among the H. influenzae isolates studied. CONCLUSIONS: Both cross colonization (between sampling sites within the same patient and between siblings) and turnover appeared to be high. PMID- 11851705 TI - Identification of enterococci and determination of their glycopeptide resistance in German and Austrian clinical microbiology laboratories. AB - OBJECTIVE: To determine exactly how German and Austrian routine laboratories perform tests for the identification of enterococci and determination of their glycopoptide resistance. METHODS: Six enterococcal test strains with different types of glycopeptide resistance (Enterococcus faecium, VanA; E. faecalis, VanA; E. faecium, VanB; E. faecalis, VanB; E. gallinarum, VanC1; E. casseliflavus, VanC2) were sent as anonymous isolates to 73 clinical microbiology laboratories (65 in Germany; eight in Austria). The participating laboratories had to identify the strains up to the species level and to determine their antibiotic susceptibilities to the glycopeptides vancomycin and teicoplanin by the test method(s) that are used daily in the corresponding laboratories. RESULTS: The analysis of the results received from 62 laboratories (56 from Germany, six from Austria) demonstrated that the most used routine method in susceptibility testing was the agar diffusion test, followed by the Etest, and the microbroth dilution procedure. The majority of participants had no difficulties in susceptibility testing with the VanA-type strains. However, the agar diffusion test was often not able to recognize clearly the VanB and VanC strains; some problems also arose with VanB isolates in the Etest. With the microbroth dilution method, the corresponding type of glycopeptide resistance was correctly determined in the majority of enterococcal test strains. Difficulties also arose in identification, especially with the VanC strains (E. gallinarum and E. casseliflavus), which were often falsely identified as E. faecium. The reasons for these errors are obviously based on the lack of important tests (such as motility and presence of a yellow pigment) in some commercially available identification test kits. PMID- 11851706 TI - Antimicrobial activity evaluations of gatifloxacin, a new fluoroquinolone: contemporary pathogen results from a global antimicrobial resistance surveillance program (SENTRY, 1997). AB - OBJECTIVE: To investigate the in vitro potency and spectrum of activity of gatifloxacin and five comparator fluoroquinolones tested against over 23 000 clinical isolates from diverse geographic and clinical sources in the Americas. METHODS: Gram-negative, Gram-positive and fastidious bacterial isolates were tested against gatifloxacin, ciprofloxacin, levofloxacin, ofloxacin, sparfloxacin and trovafloxacin using broth microdilution methods recommended by the National Committee for Clinical Laboratory Standards (NCCLS). RESULTS: Gatifloxacin demonstrated a potency and spectrum very similar to those of other new fluoroquinolones such as levofloxacin, sparfloxacin, and trovafloxacin. Gatifloxacin was particularly active against the Enterobacteriaceae (94.8% susceptible at /=98.9%), and various Staphylococcus spp. (79.2-100.0%). Trovafloxacin was the most similar comparison drug overall. CONCLUSIONS: These results indicate a potential therapeutic role for gatifloxacin that would widen the potency or spectrum of fluoroquinolones, particularly against Gram-positive species, when considering its favorable bioavailability. PMID- 11851707 TI - European survey of glycopeptide susceptibility in Staphylococcus spp. AB - OBJECTIVE: To reassess the relative potencies of teicoplanin and vancomycin following several years of clinical usage. METHODS: The glycopeptide susceptibilities of clinical isolates of staphylococci collected from 70 hospitals in 1995 were determined using NCCLS (National Committee for Clinical Laboratory Standards) methods. RESULTS: In total, 2885 isolates of Staphylococcus aureus and 1480 isolates of coagulase-negative staphylococci were collected. S. aureus was significantly less susceptible to vancomycin (MIC50 1 mg/L) than teicoplanin (MIC50 0.5 mg/L), but the reverse was the case for S. haemolyticus and S. epidermidis. No S. aureus isolate was resistant (>/=32 mg/L) to either glycopeptide, but nine isolates of coagulase-negative staphylococci had an MIC of teicoplanin of 32 mg/L. Respiratory isolates of S. aureus were less susceptible to glycopeptides than those from other sites. Staphylococci from Belgium and Italy were less susceptible to teicoplanin than isolates from other countries. CONCLUSIONS: This European survey shows that in 10 years of clinical use there have been no major changes in the susceptibility of staphylococci to the glycopeptides. PMID- 11851708 TI - MIC distribution and inoculum effect of LY333328: a study of vancomycin susceptible and VanA-type and VanC-type enterococci obtained from intensive care unit patient surveillance cultures. AB - OBJECTIVE: To determine the in vitro activity and inoculum effect of LY333328, a semisynthetic glycopeptide, against vancomycin-susceptible and vancomycin resistant enterococcal isolates. METHODS: One hundred and seventy-six enterococcal isolates (117 vancomycin-susceptible, 29 VanA-type and 30 VanC-type isolates) obtained from surveillance cultures of 139 intensive care unit patients were studied by the standard agar dilution method. Vancomycin resistance determinants were characterized by PCR. RESULTS: The activity of LY333328 was comparable (MIC range, 0.1-2 mg/L) to those of vancomycin (0.1-4 mg/L) and teicoplanin (0.06-1 mg/L) for vancomycin-susceptible isolates. LY333328 was more active (0.1-8 mg/L) than vancomycin (256 to >1024 mg/L) and teicoplanin (32-512 mg/L) against VanA-type isolates, and similar (0.2-1 mg/L) to teicoplanin (0.1 0.5 mg/L) against VanC-type isolates. The MIC distribution of LY333328 displayed a narrower range than that of vancomycin, with no clear distinction between susceptible and resistant populations. The increment in the inoculum size, from 104 to 106 CFU/spot, of susceptible isolates increased the MIC values of LY333328, vancomycin and teicoplanin by factors of 11.4, 1.6 and 3.8, respectively. The corresponding factors for LY333328 for VanA-type and VanC-type isolates were 3.5 and 6.4, respectively. CONCLUSIONS: LY333328 displays an excellent in vitro activity against vancomycin-susceptible and -resistant enterococci. Nevertheless, the inoculum size used in susceptibility tests should be carefully controlled. PMID- 11851709 TI - Evaluation of the effect of interleukin-10 on the multiplication of Mycobacterium avium complex in human macrophages and in C57BL/6 mice. AB - OBJECTIVE: To examine the effect of recombinant human IL-10 (rhIL-10) on MAC infection of human macrophages and C57BL/6 mice. METHODS: We compared rhIL-10 with the effects of the immunosuppressive drugs prednisolone and cyclosporin A, both in vitro and in vivo. RESULTS: There was no effect of rhIL-10 on the multiplication of MAC in human macrophages after 1 week of infection. In C57BL/6 mice, rhIL-10 at 2.5 or 25 m g/mouse had no additional multiplicatory effect after 3 weeks of infection, while the spleens of mice treated with prednisolone had 600% higher bacteria than controls or rhIL-10-treated mice (p0.01). CONCLUSIONS: These data suggest that rhIL-10 does not further decrease the resistance of human macrophages and C57BL/6 mice to MAC infection, whereas prednisolone leads to increased multiplication of MAC in the spleens of infected C57BL/6 mice. These results may be of interest in the context of the therapeutic use of rhIL-10 in some autoimmune disorders. PMID- 11851710 TI - Maintenance therapy for cryptococcosis in patients with AIDS after successful primary therapy: oral fluconazole (300 mg daily) versus oral itraconazole (300 mg daily). PMID- 11851711 TI - Ochrobactrum anthropi bacteremia: case report and review of the literature. PMID- 11851712 TI - Primary central nervous system aspergillosis: a case report and review of the literature. PMID- 11851713 TI - In vitro activity of gatifloxacin tested against anaerobic bacteria by reference agar dilution and Etest methods. PMID- 11851714 TI - Comparison of two agar dilution methods and three agar diffusion methods, including the Etest, for antibiotic susceptibility testing of thermophilic Campylobacter species. PMID- 11851715 TI - An adult case of fatal hemolysis induced by ceftriaxone. PMID- 11851716 TI - Nosocomial diarrhea caused by Salmonella derby infection in two patients on chemotherapy. PMID- 11851717 TI - Resistance of Salmonella and Shigella in Turkey. PMID- 11851718 TI - Clinical impact of changing to an automated blood-culture system at a small community hospital. PMID- 11851719 TI - Review of meeting: Second International Symposium of the European Study Group on Enterohemorrhagic Escherichia coli. PMID- 11851720 TI - Why is soluble intercellular adhesion molecule-1 related to cardiovascular mortality? AB - Increased plasma levels of soluble adhesion molecules are associated with an increased risk of atherothrombosis. The pathophysiological mechanisms responsible for these associations are not known. The aim of the present study was to investigate the association of soluble intercellular adhesion molecule-1 (sICAM 1) concentration and risk of cardiovascular and all-cause mortality among individuals with and without type 2 diabetes. In addition, we assessed potential pathophysiological mechanisms by which sICAM-1 may promote mortality. Six hundred and thirty-one subjects taken from a general population of the middle-aged and elderly participated in this prospective cohort study. Baseline data collection was performed from 1989 to 1992; subjects were followed until 1 January 2000. Subjects who died had higher levels of sICAM-1 than those who survived (506(164) vs. 477(162) ng mL(-1), respectively). After adjustment for age, gender and glucose tolerance status, subjects with sICAM-1 levels in the upper quartile (> or =550 ng mL(-1)) had a relative risk of cardiovascular mortality of 2.05 (95% confidence interval, 1.10-3.81) compared to subjects with sICAM-1 levels in the other quartiles. Further adjustment for classical cardiovascular risk factors or indicators of (sub)clinical atherosclerosis, endothelial dysfunction, inflammation and renal function did not materially alter this relative risk. A high sICAM-1 level was more frequent in subjects with type 2 diabetes than in subjects with a normal glucose tolerance (33.3 vs. 17.8%). Individuals with a plasma concentration of sICAM-1 higher than 550 ng mL(-1) had a cardiovascular mortality risk that was twice that of individuals with a lower concentration. Classical cardiovascular risk factors (sub)clinical atherosclerosis, endothelial dysfunction and inflammation do not explain this excess risk. PMID- 11851721 TI - PON1 L55M polymorphism is not a predictor of coronary atherosclerosis either alone or in combination with Q192R polymorphism in an Italian population. AB - BACKGROUND: The present study evaluated the role of the PON1 L55M polymorphism independently and in conjunction with the Q192R polymorphism on the risk of coronary atherosclerosis in an Italian population. MATERIALS AND METHODS: Three hundred and ninety-one subjects with significant coronary stenosis (> 50%) (coronary artery disease-positive; CAD+), 196 subjects with normal coronary arteries (< 10% stenosis) (CAD-) and 178 healthy controls were screened using a combination of polymerase chain reaction and restriction enzyme digestion. RESULTS: In the pooled population, the frequencies of L and M alleles were 0.63 and 0.37, respectively; the most common haplotypes were QQ/LM (24.2%) and QR/LL (21.8%) and a strong linkage disequilibrium between L/55 and R/192 alleles was observed (D' = -0.91; P < 0.0001). CAD+ subjects did not show any significant differences in the distribution of PON1-55 genotypes as compared to CAD- subjects and population controls (chi2 = 1.5, P = 0.8). After controlling for other risk factors, the low-concentration M allele was not associated with a significant change of CAD risk (OR 1.02; 95% CI 0.80-1.29; P = 0.87). Moreover, the L55M polymorphism did not show any interaction with other risk factors such as smoking, diabetes, hypertension, low levels of high-density lipoprotein (HDL) or high ratios of low-density to high-density lipoproteins. The combination of L55M with the Q192R polymorphism did not show any effect on CAD risk. However, a marginal decrease in myocardial infarction risk was detected when QQ/MM carriers (OR 0.51; 95% CI 0.26-0.99; P = 0.048), but not LL/RR carriers, were compared with subjects not homozygous for an L or R allele. CONCLUSIONS: These findings did not indicate a major effect of the PON1 L55M polymorphism, either alone or in combination with the Q192R polymorphism, on CAD risk. Additional studies are needed for a better evaluation of the role of the 55/192 PON1 genotypes in combination on myocardial infarction risk. PMID- 11851722 TI - Cardiac adaptation to intensive training in prepubertal swimmers. AB - BACKGROUND: Despite the increasing involvement of child athletes in intensive training regimens, little is known about the influence of such training on autonomic regulation and cardiac structure and function. PATIENTS AND METHODS: Twenty-five highly trained (12-14 h weekly for at least 4 years) swimmers (aged 11.9 +/- 1.6 years; 15 males, 10 females) and 20 non-training normal children who served as controls (aged 11.3 +/- 0.6 years; 14 males, 6 females) were studied. Heart rate variability analysis in the time and frequency domains was performed on 15 min resting heart rate acquisitions. Left ventricular morphology and systolic function was studied with two-dimensional guided M-mode echocardiography. The transmitral flow velocity profile was assessed with pulsed Doppler. Parameters measured included the peak early (E) and peak late (A) transmitral flow velocity and their ratio (E/A). Left atrial (LA) volumes were determined at mitral valve (MV) opening (maximal, Vmax), at onset of atrial systole (P wave of the ECG, Vp), and at MV closure (minimal, Vmin) from the apical 2- and 4-chamber views, using the biplane area-length method. LA systolic function was assessed with the LA active emptying volume (ACTEV) = Vp-Vmin and the LA active emptying fraction (ACTEF) = ACTEV/Vp. RESULTS: Average NN (967.1 +/ 141.8 vs. 768.4 +/-85.6 ms, P < 0.0001), logSDNN (1.89 +/- 0.14 vs. 1.80 +/- 0.17 ms, P < 0.05), logPNN 50% (1.66 +/- 0.23 vs. 1.46 +/- 0.35, p < 0.05), and logHF power (3.13 +/- 0.32 vs. 2.95 +/- 0.26 ms2, p < 0.05) were greater in swimmers than in controls. Left ventricular end-diastolic diameter was greater (32.3 +/- 3.3 vs. 29.5 +/- 3.3 mm m(-2), P < 0.02) in swimmers than in controls, whereas the left ventricular septal (5.9 +/- 1 vs. 5.6 +/- 0.8 mm m(-2), P = NS) and posterior wall thickness (5.7 +/-0.9 vs. 5.4 +/- 0.8 mm m(-2), P = NS) were similar in the two groups. The E/A ratio was greater (2.2 +/- 0.49 vs. 1.78 +/- 0.36, P < 0.003) whereas the A velocity was lower (0.41 +/- 0.09 vs. 0.50 +/- 0.13 m s(-1), P < or = 0.02) in swimmers than in controls. Vmax was greater (18.6 +/-4.8 vs. 14.9 +/-5.3 cm m(-2), P < 0.03), whereas ACTEF was lower (36 +/- 12% vs. 44.2 +/- 12%, P < 0.04) in swimmers than in controls. CONCLUSION: Cardiac adaptation to intensive training in prepubertal swimmers includes vagal predominance, a mild increase in left ventricular dimensions without significant changes in septal or posterior wall thickness, and increased LA size associated with depressed LA systolic function. Evaluation of LA size and systolic function may contribute to a better understanding of the characteristics of the 'athlete's heart' in children and to the differential diagnosis between left ventricular adaptive and pathologic changes. PMID- 11851723 TI - Plasma homocysteine, methylenetetrahydrofolate reductase gene polymorphism and carotid intima-media thickness in Italian type 2 diabetic patients. AB - BACKGROUND: Moderately elevated levels of homocysteine have been associated with an increased cardiovascular risk in type 2 diabetic patients. The role of methylenetetrahydrofolate reductase gene polymorphism is less clear. MATERIALS AND METHODS: We investigated the contribution of plasma homocysteine levels and the methylenetetrahydrofolate reductase gene polymorphism to the variability of carotid intima-media thickness in 124 consecutive Italian patients with type 2 diabetes mellitus. Fasting plasma homocysteine was measured by high-pressure liquid chromatography with an electrochemical detector; methylenetetrahydrofolate reductase genotypes were determined by polymerase chain reaction and restriction enzyme digestion. The carotid intima-media thickness was evaluated with high resolution B-mode ultrasonography. RESULTS: Age, creatinine and plasma homocysteine levels showed a positive correlation with mean carotid intima-media thickness values, but only age and creatinine levels were still associated with mean carotid intima-media thickness values in the multivariate analysis. Plasma homocysteine levels were significantly higher in the patients bearing the 677T/677T genotype of the methylenetetrahydrofolate reductase polymorphism; mean carotid intima-media thickness values were not different in the three different methylenetetrahydrofolate reductase genotypes. CONCLUSION: In 124 Italian patients with type 2 diabetes mellitus, basal levels of plasma homocysteine, as well as methylenetetrahydrofolate reductase gene polymorphism, did not explain the variability of mean carotid intima-media thickness. PMID- 11851724 TI - Effects of deoxycholate on human colon cancer cells: apoptosis or proliferation. AB - BACKGROUND: Deoxycholic acid has long been attributed as a tumour promoter in the colon. It exerts its growth-related actions in a phorbol ester-like manner, by stimulating protein kinase C. The aim of this study was to investigate the effect of deoxycholic acid on proliferation and apoptosis in the colon, by exposing colon cancer cells to it in increasing concentrations. METHODS: Human colon cancer cells (Caco-2 and HT-29) were treated with deoxycholate or its two structural isomers, 3-beta-12-alpha-dihydroxy-5-beta-cholan-24-oic acid and 3 alpha-12-beta-dihydroxy-5-beta-cholan-24-oic acid. Proliferation was evaluated by cell counting, and apoptosis by estimating percentage cell survival and assessment of nuclear morphology. RESULTS: Within the concentration range of up to 20 microM, deoxycholate stimulated growth of both human colon cancer cell lines. Its growth-promoting effect was abolished after inhibition of protein kinase C. At concentrations above 100 microM, deoxycholate induced apoptosis in both cell lines. Epimers of deoxycholate were significantly less potent in stimulating growth. CONCLUSION: Low-dose deoxycholate stimulates colon cancer cell proliferation while > 100 micromol L(-1) of this secondary bile acid induces apoptosis in colon cancer cells. Deoxycholate might promote the likelihood of malignant transformation by increasing epithelial cell turnover in the colon. PMID- 11851725 TI - Oxysterols induce interleukin-1beta production in human macrophages. AB - BACKGROUND: Oxysterols are biologically active molecules generated during the oxidation of low-density lipoprotein or formed enzymatically in vivo. In the atherosclerotic plaque newly recruited macrophages may be exposed to oxysterols present in the plaque. How these oxysterols affect the expression and secretion of inflammatory cytokines such as interleukin-1beta (IL-1beta) in macrophages is not known. Therefore the aim of the present study was to investigate how oxysterols regulate the expression and secretion of IL-1beta in human monocyte derived macrophages. METHODS: The IL-1beta messenger RNA (mRNA) expression was analysed by reverse transcription-polymerase chain reaction, and the IL-1beta protein secretion was measured by enzyme-linked immunosorbent assay. RESULTS: A significant, dose-dependent increase in the secretion of IL-1beta was given by 25 hydroxycholesterol without the addition of lipopolysaccharide (LPS). At a concentration of 2.5 microg mL(-1) this increase was similar to that obtained by endotoxin (LPS, 1 microg mL(-1)). A transient increase in IL-1beta mRNA expression was found in macrophages incubated with 25-hydroxycholesterol compared with untreated controls. In addition, 25-hydroxycholesterol dramatically increased the IL-1beta secretion induced by LPS. At a concentration of 5 microg mL(-1) of 25-hydroxycholesterol the LPS-induced IL-1beta secretion was increased by about 25-fold. A similar tendency, but not so consistent, was found for 27 hydroxycholesterol. CONCLUSIONS: Our results show that oxysterols, and 25 hydroxycholesterol in particular, may modulate the inflammatory response in human macrophages. Consequently the presence of oxysterols in atherosclerotic tissue may dramatically influence the effect of inflammation. PMID- 11851726 TI - Increased serum interferon alpha in HIV-1 associated lipodystrophy syndrome. AB - BACKGROUND: A syndrome of lipodystrophy (LD) associated with peripheral lipoatrophy and central/visceral adiposity has been reported in HIV-1-infected patients treated by combined antiretroviral therapy (ART). Lipid metabolism is partly regulated by both steroid hormones and cytokines and we have previously reported that dyslipidaemia in LD-positive men is correlated to an increase in cortisol : DHEA ratio (Christeff et al., AIDS 1999;13:2251). In this study we questioned whether it is also related to cytokine perturbations. MATERIALS AND METHODS: A cross-sectional study was performed on 42 HIV-1-positive men on ART, 27 of whom had symptoms of LD, defined by computed tomography scan. Serum cytokines (IFN-alpha, TNF-alpha, sTNF-RI, sTNF-RII, IL-6, IL-1beta and IL-2) and lipids [cholesterol, triglycerides (TG) and their subclasses], and apolipoproteins (Apo), were determined. RESULTS: Serum IFN-alpha was markedly increased in LD-positive compared with LD-negative men and controls. IL-6 and TNF alpha concentrations were also significantly elevated in HIV-positive men compared to controls but the levels of these cytokines did not differ between the two groups of patients. A significant positive correlation was found between accumulation of IFN-alpha and increased levels of cholesterol, TG, VLDL cholesterol, VLDL TG, ApoB and ApoB-ApoA1 ratio. A multivariate forward-performed analysis revealed that IFN-alpha is the best marker for lipid perturbations associated to LD, followed by insulin and cortisol : DHEA ratio. CONCLUSIONS: This study demonstrates an association between serum IFN-alpha and lipid alterations in LD-positive men. The concomittant action of IFN-alpha and cortisol : DHEA ratio is probably one of the mechanisms responsible for hyperlipidaemia in LD syndrome. PMID- 11851727 TI - d-penicillamine reduces serum oxidative stress in Alzheimer's disease patients. AB - BACKGROUND: Several lines of evidence address the emerging role for copper in Alzheimer's disease (AD) for sustaining oxidative mechanisms. Studies indicate that peripheral markers of oxidative stress in AD patients could be informative about the pathophysiology of this brain condition. Here, we present a pilot study examining the efficacy of the copper-chelating agent d-penicillamine in reducing oxidative stress in AD patients. DESIGN: Serum levels of copper sampled in AD patients and healthy controls indicate a copper homeostasis imbalance in AD. On this basis, 34 AD patients were enrolled in a 6-month, double-blind, placebo controlled trial with the copper d-penicillamine-chelating agent. Nine patients for each group completed the trial. Oxidative stress, trace metals and clinical parameters were evaluated. RESULTS: At the start of the study (t0) total peroxides and copper serum content of AD patients were higher (P < 0.0001, P < 0.0001, respectively) and antioxidants were lower (P < 0.05) than in healthy controls. Copper and peroxides were correlated in the AD population (Pearson's r = 0.61, P < 0.001). After treatment with d-penicillamine, the extent of oxidative stress (P < 0.05) was decreased, but no difference was observed in the rate of cognitive decline. CONCLUSION: Data from this pilot study suggest that copper could play a role in the production of peroxides in AD, and that d-penicillamine has an effect in reducing oxidative damage, however, results are still inconclusive in terms of drug efficacy on the clinical progression of AD. Studies with larger cohorts are needed to elucidate the real effectiveness of d penicillamine treatment in AD. PMID- 11851728 TI - Secretases as targets for drug design in Alzheimer's disease. AB - Alzheimer's disease accounts for the majority of dementia in the elderly. Worldwide, approximately 20 million people are suffering from this devastating disease, with no effective treatment currently available. For efficient drug design, it is important to identify the molecular mechanisms underlying the pathology of the disease. An invariant feature in the pathology of Alzheimer's disease is the amyloid-beta peptide. Amyloid-beta is produced by endoproteolytic cleavage of the amyloid precursor protein by beta- and gamma-secretase. In the past 2 years, the protein responsible for beta-secretase activity has been isolated and researchers are close to identifying gamma-secretase. These recent achievements in Alzheimer's disease research have provided helpful tools for the development of therapeutics. PMID- 11851730 TI - Current evidence for the use of botulinum toxin type A in the management of children with cerebral palsy: a systematic review. AB - Management of children with cerebral palsy (CP) is the focus of considerable resources in many countries, so that evaluation of the efficacy for new and established treatments is imperative. Botulinum toxin type A (BTX-A) is a relatively new method of spasticity management in children with cerebral palsy. It has been the focus of extensive research since its application to cerebral palsy 10 years ago. In a systematic review relating to the management of the lower limb in cerebral palsy 156 papers were identified. These were categorized according to Sackett and the World Health Organisation International Classification of Impairments, Disabilities and Handicaps model. We identified 10 randomized trials evaluating the use of BTX-A in the lower limb in children with cerebral palsy in a systematic review. A meta-analysis showed the pooled risk difference between BTX-A and placebo in three trials was 0.25 (95% CI 0.13, 0.37) and 0.23 (95% CI -0.06, 0.53) for two trials of BTX-A and casting using the physicians rating scale. These represent moderate treatment effects that are dosage-dependent. Outcomes were also compared for function in five studies. The type of evidence for BTX-A was graded by each treatment indication and directions for future research were then drawn from the available evidence. PMID- 11851731 TI - Botulinum toxin type A and other botulinum toxin serotypes: a comparative review of biochemical and pharmacological actions. AB - Botulinum toxin type A is an important therapeutic agent for the treatment of movement and other disorders. As the clinical uses of botulinum toxin type A expand, it is increasingly important to understand the biochemical and pharmacological actions of this toxin, as well as those of other botulinum toxin serotypes (B-G). Botulinum neurotoxin serotypes exhibit differences in neurotoxin complex protein size, percentage of neurotoxin in the activated or nicked form, intracellular protein target, and potency. These properties differ even between preparations that contain the same botulinum toxin serotype due to variations in product formulations. As demonstrated in preclinical and clinical studies, these differences result in a unique combination of efficacy, duration of action, safety, and antigenic potential for each botulinum neurotoxin preparation. PMID- 11851732 TI - Botulinum toxin type A management of spasticity in the context of orthopaedic surgery for children with spastic cerebral palsy. AB - Cerebral palsy is the most common cause of physical disability affecting children in developed countries. Although cerebral palsy is, by definition, a 'static encephalopathy' the associated musculoskeletal pathology is progressive and current definitions are therefore somewhat inadequate. Understanding the stages of the musculoskeletal pathology is fundamental to understanding current management strategies, including spasticity management, strengthening programmes and deformity correction by orthopaedic surgery. In this review, a number of new management strategies are described, in which spasticity management by intramuscular injections of botulinum toxin type A and deformity correction, by orthopaedic surgery, are combined. PMID- 11851733 TI - Spasticity versus strength in cerebral palsy: relationships among involuntary resistance, voluntary torque, and motor function. AB - Despite the lack of consensus of the role of spasticity in the observed motor disability in cerebral palsy (CP), alleviation of spasticity remains a primary focus in the clinical management of these patients. The purposes of this study were to: (1) quantify voluntary torque and passive resistance across speeds in the hamstrings and quadriceps muscle groups with respect to the presence of stretch responses and/or passive muscle stiffness in patients with CP compared to age-related children without disability, and (2) relate these parameters to each other and to functional performance, as measured by the Gross Motor Function Measure (GMFM), in CP. Included were 23 subjects with CP, sub-grouped by the presence or absence of stretch responses as determined by electromyography, and 9 subjects without CP. Results indicated that peak torque was considerably greater in the comparison group than for each of the CP groups and resistance was greater in the CP group with spasticity compared to the nonspastic CP group in both muscles at all speeds. Stiffness differed between the spastic CP group and the comparison group only for the quadriceps at the fastest speed. Higher passive resistance torque and stiffness were correlated with decreased voluntary torque, particularly for the antagonists, and with lower GMFM scores. In conclusion, strength and motor function are related to the magnitude of resistance torque and stiffness in CP, although the small amount of variance explained reinforces the multidimensional nature of this disorder, and the challenges inherent in managing it. PMID- 11851734 TI - The effect of botulinum toxin type A on the functional ability of the child with spastic hemiplegia a randomized controlled trial. AB - It has been demonstrated that botulinum toxin type A (BTX-A) injections reduce spasticity and improve muscle growth in children with spasticity. It has been postulated that BTX-A allows the learning of more normal movement patterns. The aim of this study was to measure the effect of this treatment on functional ability, as measured by the Gross Motor Function Measure (GMFM), in children with spastic hemiplegic cerebral palsy. Children of 3--13 years and meeting the selection criteria were randomly allocated to the control or injection group using a matched pair design. A match constituted a child within 6 months of age with the same Modified Ashworth Score (MAS) for the gastroc-soleus and within 10% of the same goal scores on the Gross Motor Function Measure. Twelve matched pairs were enrolled. Outcomes were measured on enrolment and at 1, 3 and 6 months post injection. The time course of the response to BTX-A was assessed with measurements of the MAS, dynamic range of motion (R1) and static muscle length (R2). Motor function was assessed using the 88-item GMFM and parental satisfaction with a 10-point visual analogue scale. Within pair comparisons of the GMFM using the Wilcoxon signed rank test indicated that the treatment group made significantly greater gains than controls at 3 months (P=0.02) with even greater differences seen at 6 months (P=0.004). Using parametric statistics, the intrapair difference in proportional change of GMFM increased from 35% (4 to 65) at 3 months to 52% (17--87) at 6 months. Response to injection was confirmed by a decrease in MAS in the treatment group and very little change in controls. This difference was significant (P=0.002) at 3 months and was attenuated but still significant (P=0.016) at 6 months; the difference in proportional change decreased from 44% at 3 months to 22% at 6 months. Changes in R1 reflected those of MAS in the treatment group and deteriorated significantly over the study period in controls. Parents of children in the treatment group were more satisfied than controls, but satisfaction scores did not correlate with changes in function or technical outcomes suggesting that this may be a placebo effect. The changes in GMFM correlated with changes in technical outcomes at 3 months, suggesting a causal relationship. The intrapair differences in GMFM continued to increase even after the local response to injection had started to wane. PMID- 11851735 TI - Treatment of functional limitations at the knee in ambulatory children with cerebral palsy. AB - The gait of ambulatory children with cerebral palsy frequently involves abnormal knee motion. Spasticity, muscle contracture formation, impairments of motor control, weakness, balance deficits, and extrapyramidal motions can all contribute to the functional limitations imposed at the knee. Careful clinical evaluation of the child and their gait must be performed in order to determine the best individual course of treatment. Often, three-dimensional motion analysis with assessment of muscle activity and force is necessary to completely assess the complexities of gait. Several typical gait patterns have been described involving the knee, including 'jump knee', 'crouch', 'true equinus', 'apparent equinus', 'recurvatum' and 'stiff knee' gait. Each of these gait patterns is defined here and discussed using case examples. These typical gait patterns are usually accompanied by involvement at the hip and ankle and may be combined with transverse plane rotational abnormalities. Treatment options such as rehabilitation (physiotherapy, casting, strengthening, and/or orthoses), spasticity management (intramuscular injections of phenol, alcohol, and botulinum toxin type A) and orthopaedic approaches are discussed for each entity. PMID- 11851736 TI - A randomized study of combined botulinum toxin type A and casting in the ambulant child with cerebral palsy using objective outcome measures. AB - It is recognized that objective gait analysis is of great value in planning a multilevel botulinum toxin type A (BTX-A) treatment. After BTX-A treatment, objective outcome measures can provide new and interesting information for each individual child with cerebral palsy (CP). Moreover, by studying group results, we may evaluate our treatment hypotheses. The present prospective study attempts to document the effect of integrated multilevel BTX-A treatment on objective gait parameters and to define the optimal strategy for the combined treatment of BTX-A with casting in children with cerebral palsy. Objective three-dimensional gait analysis (3DGA) data were collected pre- and 2 months post-treatment, in two randomized patient groups: a first group of 17 children treated with lower leg casting prior to BTX-A injections, and a second group of 17 patients who received casting immediately after injections. The present study demonstrates that improved gait can be achieved after a multilevel BTX-A treatment, combined with casting, using a set of 90 gait parameters. The most pronounced improvement was seen at the ankle joint. The results in the knee, hip and pelvis imply that multilevel treatment of the child with CP should start at an early age, in order to prevent development of muscle contractures. Slightly more pronounced benefits, mainly in the proximal joints, were seen for the children who were casted after injections as compared to the children who were casted before injections. PMID- 11851738 TI - Classification of gait patterns in spastic hemiplegia and spastic diplegia: a basis for a management algorithm. AB - Classifications of gait and postural patterns in spastic hemiplegia and spastic diplegiia are presented, based on the work of previous authors. The classifications are used as a biomechanical basis, linking spasticity, musculoskeletal pathology in the lower limbs, and the appropriate intervention strategies. The choice of target muscles for spasticity management, the muscle contractures requiring lengthening and the choice of orthotics are then linked to the underlying gait pattern. PMID- 11851737 TI - Single event multilevel botulinum toxin type A treatment and surgery: similarities and differences. AB - The present study attempts to provide objective evidence of two treatment options for children with cerebral palsy (CP): multilevel botulinum toxin type A (BTX-A) injections and multilevel surgery. The purpose of the study was to clarify the differences and the similarities, and common treatment principles of both treatment strategies. Objective three dimensional gait analysis data were studied retrospectively in two patient groups pre- and post-treatment (randomly selected from a group of children that were treated between 1998 and 1999). In the first group, 29 children with CP were managed with BTX-A injections according to an integrated multilevel approach (Molenaers et al., 1999a). A second group of 23 children with CP were managed by a more traditional single event multilevel surgery, also according to an integrated approach. Our aim was to evaluate the differences as well as the similarities between both patient groups, using a set of 56 parameters selected from three-dimensional gait analysis. The unifying concept between management with BTX-A injections and orthopaedic surgery was the adoption of a multilevel approach at one session. The groups demonstrated considerable differences with respect to age, pretreatment condition and amount and level of improvement after treatment. The children who received BTX-A were typically younger, and showed primary gait problems in the distal joints, whereas the children who underwent surgery demonstrated a higher frequency of gait deviations in the transverse plane and had more complications. Although the benefit of both treatments was confirmed by the present study, a difference in the amount and level of improvement was also demonstrated. In conclusion, these treatment modalities should be regarded as complementary rather than mutually exclusive treatments, with both calling for an integrated approach. PMID- 11851739 TI - The effect of botulinum toxin type A and a variable hip abduction orthosis on gross motor function: a randomized controlled trial. AB - Hip displacement is the second most common deformity after equinus in children with cerebral palsy (CP), and may result in dislocation, pain, fixed deformity and loss of function. We studied the combined effects of intramuscular injections of botulinum toxin type A (BTX-A) to the adductors and hamstrings and a variable hip abduction orthosis (SWASH), on gross motor function, hip displacement and progression to surgery, in a randomized clinical trial. Thirty-nine children, with bilateral spastic cerebral palsy, and mean age 3 years + 2 months (range 1 year+7 months--4 years +10 months) entered the trial. Gross Motor Function Classification System (GMFCS) levels were as follows: one child was level II, 11 were level III, 13 were level IV and 14 were level V. After concealed randomization, 20 were allocated to the control group and 19 to the intervention group. Thirty-five children completed the follow up at 1 year. The novel intervention group received up to 4.0 U BOTOX/kg/muscle, 16 U/kg/body weight every 6 months plus the use of a SWASH brace. The control group received clinical best practice comprising physiotherapy but no hip abduction bracing. Both groups showed improvements in total Gross Motor Function Measure (GMFM) score [mean 6.0% BTX-A group; 6.1% Control; 95% CI -- 6.7, 6.5 (NS)], however, there was no additional treatment effect for the study group. There were similar improvements on GMFM goal scores and GMFM-66 scores, but again no additional treatment effect was observed. Multiple regression of change in total GMFM by GMFCS classification for each group showed greater improvement in the total scores from baseline in the BTX-A/SWASH treated group than the control group. In the first year, nine children (two in the intervention group and seven in the control group) required soft tissue surgery because of progressive hip migration in excess of 40%. A longer-term follow up of a larger cohort may be required to determine the effect of the combined treatment on hip displacement. PMID- 11851740 TI - Medium-term functional benefits in children with cerebral palsy treated with botulinum toxin type A: 1-year follow-up using gross motor function measure. AB - One of the main goals when treating spasticity is to relieve pain and improve function. Intramuscular injection of botulinum toxin type A (BTX-A) has gained widespread acceptance in the treatment of spastic cerebral palsy. Several studies have clearly shown the short-term functional benefit of BTX-A treatment. Information is limited, however, on the efficacy of medium and long-term regimens, using repeated injection of BTX-A. The aim of the present open-label, prospective study was to evaluate functional outcome in children with spastic cerebral palsy after 1 year of treatment with BTX-A, using the Gross Motor Function Measure (GMFM) as a validated outcome measure. Patients (n=25, age 1.5- 15.5 years) were treated with BTX-A for adductor spasm (n=12) or pes equinus (n=13). The local effect was evaluated using passive range of motion and modified Ashworth Scale. Apart from a significant improvement in joint mobility and reduction of spasticity compared to pretreatment values (P < 0.01), we demonstrated a significant improvement of gross motor function after 12 months of treatment, with a median gain of 6% in total and goal scores (P < 0.001). An increase in GMFM scores was particularly evident in younger and moderately impaired children (Gross Motor Function Classification System level III). Whether the observed improvement in gross motor function in children with cerebral palsy is specifically related to therapy with BTX-A or represents at least in part the natural course of motor development still needs clarification. PMID- 11851741 TI - Spasticity management in the child with spastic quadriplegia. AB - In children with spastic quadriplegia, also described as 'whole body involvement', spasticity can interfere with motor function, contributes to the development of deformities and adversely impacts on care, positioning, and comfort. In this population, spasticity interventions address goals such as improving comfort, reducing pain, easing the burden of carers, slowing the progression of musculoskeletal deformities and perhaps improving function. Children with severe diplegia are distinguished from those with quadriplegia by their ability to ambulate, as well as by a greater emphasis being placed on functional motor goals even though similar treatment modalities are often employed to manage spasticity. The many treatment options currently available include, but are not limited to, botulinum toxin type A, phenol neurolysis, oral medications, intrathecal baclofen, selective dorsal rhizotomy, and orthopaedic surgery. The integration of these treatment modalities can help to optimize the overall care and function for a child with spastic quadriplegia or severe diplegia. However, the development of a management programme is complex and needs to take into account many factors, including age, weight and nutritional status, rate of progression of musculoskeletal deformities, developmental potential, comorbid conditions, current functional status and prognosis, and family and patient treatment goals. Children with marked spasticity are likely to benefit from a combination of interventions, rather than a single treatment modality. Because of these complexities, management should be planned and coordinated by a multidisciplinary team of medical and allied health professionals which recognizes the central role of the family in all decisions. Once the special characteristics of the child with spastic quadriplegia and the various treatment options are understood, outcomes can be maximized. PMID- 11851742 TI - Management of the upper limb with botulinum toxin type A in children with spastic type cerebral palsy and acquired brain injury: clinical implications. AB - The aim of this article is to describe our clinical experience in treating muscle imbalance in 49 children with spastic upper extremity involvement. We discuss four cohorts of children treated with botulinum toxin type A (BTX-A), each with different treatment objectives. In the first group, 27 children were treated for functional improvement and, of these, 23 had a positive effect, while four had no objective benefit. In the second group, eight children were treated for purposes of presurgical planning; of these, four were referred for surgery, three continued with serial treatment and one child did not benefit from injection. The third group comprised six children who were treated to improve posture and care: in this group, four children demonstrated clear benefit and two children lost some function subsequent to injection. Finally, a fourth group of seven children were treated after acquired brain injury (three with severe tetraplegia, four with hemiplegia). In this group, all children experienced spasticity relaxation and two children with hemiplegia also gained functional benefit. In terms of adverse events, deterioration of upper extremity function was poorly tolerated but limited to the first 1--3 weeks postinjection. Grip strength or thumb grip were diminished if too high doses were used. Overall, our results with BTX-A were rewarding in children with no fixed contracture, good motor learning capacity and high motivation to train. Additionally, BTX-A treatment has proven valuable for counteracting spasticity in children with acquired brain injury. This treatment modality may not, however, be an appropriate treatment option for all children with severe upper extremity spasticity, due to the shorter duration of effect and the potential reduction in functional abilities seen in this cohort. In all cases, the selection of muscles to be treated needs careful clinical assessment. Dynamic EMG analysis should be performed whenever required to aid muscle selection, especially in children with spasticity combined with dystonia. Evaluation of M-responses suggests that for the forearm muscles, doses of BTX-A above 1.5 U/kg/muscle should not be used. PMID- 11851743 TI - Botulinum toxin type A injections in the spastic upper extremity of children with hemiplegia: child characteristics that predict a positive outcome. AB - Evidence is increasing to suggest that botulinum toxin type A (BTX-A) plays a role in the management of upper extremity spasticity in the paediatric population. However, little information is available on the clinical characteristics of the child that predict a response to this intervention. Our research group previously published a randomized controlled trial demonstrating that BTX-A injection improves function of the upper extremity of children with spastic hemiplegia. In the present paper, we evaluate the child characteristics that predict a positive response to the BTX-A injections in the randomized treatment group. The treatment group was divided into positive functional responders and nonresponders using a cut score of a change of 10 points on the Quality of Upper Extremity Skills Test (QUEST). A two-way analysis of variance procedure was done comparing the following baseline characteristics: function as scored on the QUEST and the Pediatric Evaluation of Disability Inventory (PEDI) self-care domain, grip strength, upper extremity spasticity and age. Grip strength was significantly higher in responders with a P-value of 0.001. Young age approached significance with a P-value of 0.05. Correlation of change scores on the QUEST with baseline characteristics in the treatment group yielded similar results. BTX-A causes a reduction in spasticity and strength; we postulate that if the hand is weak initially, BTX-A can decrease hand function. Two case reports are presented that highlight the importance of grip strength and age. PMID- 11851744 TI - Management of upper limb dysfunction in children with cerebral palsy: a systematic review. AB - Effective use of the upper limb can impact on educational outcomes, participation in activities of daily living and vocational options for many children with cerebral palsy (CP). This article presents the results of a systematic review of the literature on the management of upper limb dysfunction in children with CP. The range of management options includes therapies such as physiotherapy, occupational therapy, neurodevelopmental therapy and conductive education; peripheral splinting and casting; focal or generalized pharmacotherapy; and surgery to improve upper limb function or correct deformity. A literature search identified 60 papers, of which four were randomized controlled trials and 44 were prospective studies with objective outcome measures. Principal studies undertaken for each type of treatment and the efficacy of the different types of treatment were critically evaluated. In addition, the current level of evidence for each study was evaluated according to Sackett's (1989) model and ICIDH-2 classification. A close examination of two relatively new treatments for upper limb spasticity, constraint induced movement therapy and botulinum toxin type A (BTX-A) was conducted with reference to more extensive data on the efficacy of BTX-A in the lower limb. PMID- 11851745 TI - Outcome measurement of effectiveness of botulinum toxin type A in children with cerebral palsy: an ICIDH-2 approach. AB - The ICIDH-2 serves as a useful framework for differentiating measurement by dimensions of the disabling process. Such differentiation is important to achieve more valid measurement of health related outcomes. We have attempted to examine one intervention, treatment with botulinum toxin type A, for one patient population, children with cerebral palsy, and to describe the outcome measures used in the evaluation of that intervention using this evolving classification system. This process supports the concept that measurement of health outcomes should focus on the nature and extent of functional limitations in physical, social and psychological domains. The selection of measurement outcomes must be determined not only by the requirements of the scientific process but also by the needs of the patients who are the intended beneficiaries of the intervention. PMID- 11851746 TI - Proportional change: an additional method of reporting technical and functional outcomes following clinical interventions. AB - One of the major challenges in disability management research is to express results in a manner that can be generalized to subjects with varying degrees of disability. Absolute measurements of change are often dependent on initial characteristics, in which case they can only be generalized to subjects with the same characteristics. We define proportional change as the ratio of change to the maximum possible or targeted change. It can be assessed in any situation where a maximum possible or targeted change is definable. Its estimated value will be sensitive to the choice of denominator. Subjective assessments, such as those measured with Likert scales, are naturally expressed as proportional change with the denominator being set by the subject. Denominators may also be determined objectively by physical limitations or, less desirably, by the measurement tool. Where there is no readily or objectively determinable denominator, they should be chosen for each subject in advance of the intervention according to carefully specified criteria. Proportional change is proposed, as an adjunct to the reporting of absolute measures of change following therapeutic interventions, as a means of expressing change in a manner that is both individualized and generalizable. PMID- 11851747 TI - The measurement of health-related quality of life (HRQL) in children with cerebral palsy. AB - The measurement of health-related quality of life (HRQL) in children with cerebral palsy is part of an emerging discipline. Theoretical models of disability and chronic illness developed by the World Health Organization, the National Center for Medical Rehabilitation Research and others are being adapted for children. Development of HRQL measures in paediatrics lags behind the work completed with adults due to challenges unique to children, such as what domains to measure, whose perspective to address and the developmental changes inherent to children. The descriptive HRQL studies of children with moderate to severe cerebral palsy and comparisons of children with quadriplegia vs. diplegia and hemiplegia are presented. Consistent with the functional measures, no significant difference in HRQL by treatment was documented in children with spastic diplegia participating in a randomized clinical trial of selective dorsal rhizotomy. HRQL measures that are specifically tailored to cerebral palsy need to be developed. Generic and individualized measures of HRQL are currently available and while limited, may be useful for evaluating the effect of different treatments for spasticity on the quality of life in children with cerebral palsy. PMID- 11851748 TI - Botulinum toxin type A in the management of equinus in children with cerebral palsy: an evidence-based economic evaluation. AB - INTRODUCTION: The aim of this study was to compare two methods of conservative management of calf spasticity and equinus gait--intramuscular injection of botulinum toxin type A (BTX-A) and serial casting. An economic evaluation framework was adopted to assess whether BTX-A offers value for money in the management of equinus gait due to calf spasticity in children with cerebral palsy. Short- to medium-term health care costs and outcomes were estimated for the comparison. This study was embarked upon to provide clinical and economic data as part of an application to the Pharmaceutical Benefits Advisory Committee (PBAC) for the reimbursement of BOTOX by the Australian Commonwealth Government. This is the primary mechanism for reimbursement of pharmaceuticals in Australia, as they are not routinely reimbursed through health insurance companies. The perspective of the analysis was that of the Australian health system. METHODOLOGY: Randomized controlled trials (RCTs) exist comparing one treatment cycle of BTX-A with serial casting (Corry et al., 1998; Flett et al., 1999). A long-term prospective study provided data on multiple cycles of BTX-A treatment in a more naturalistic setting (Boyd et al., 1999). A simple economic modelling approach was used to establish resource utilization by treatment arm. Only direct medical costs were considered (BTX-A, medical personnel time and medical consumables). MAIN MEASURES: Clinical efficacy was obtained from the randomized controlled trials (Corry et al., 1998; Flett et al., 1999). Patient/parent preference was obtained from long-term follow-up (Corry et al., 1998) and a preference questionnaire (Flett et al., 1999). Australian treatment patterns and patient demographics were obtained from the naturalistic study (Boyd et al., 1999). RESULTS: The RCTs demonstrated equivalent efficacy of BTX-A and serial casting; however, with BTX-A the effect lasted longer and was clearly the preferred treatment. For patients with hemiplegia the costs of an episode of treatment with BTX-A or serial casting are ($AUD) $595 and $435, respectively, and thus the additional costs associated with BTX-A are $160. The corresponding costs for patients with diplegia are $1045 for BTX-A treatment and $870 for serial casting and thus the additional cost associated with BTX-A is $175. With an overall treatment duration of 3.7 years and an average treatment interval of 10 months, patients would receive an average of 5.4 treatments. Thus, for patients with hemiplegia the total additional cost, discounted at 5% annually, for BTX-A is $793. For patients with diplegia the total additional cost for BTX-A is $867. CONCLUSIONS AND CLINICAL INTERPRETATIONS: BTX-A is an effective, safe and acceptable treatment modality and is associated with only a modest increase in direct medical costs per child per year. BTX-A can be considered a valuable and cost-effective treatment in the conservative management of equinus due to calf spasticity in children with cerebral palsy. This conclusion is supported by the acceptance of the Pharmaceutical Benefits Advisory Committee (PBAC) recommendation that BOTOX should attract a full Government subsidy in Australia. PMID- 11851750 TI - Continuous infusion of porcine factor VIII in patients with haemophilia A and high-responding inhibitors: stability and clinical experience. AB - A multicentre retrospective survey was conducted to assess the efficacy and side effect profile of porcine factor VIII (pFVIII:C) given by continuous infusion (CI) to patients with congenital haemophilia A and inhibitors. Twenty-nine episodes in 18 patients were treated by CI of pFVIII:C. Efficacy was graded as good in 79% of infusions and fair in 17%. There was a failed response in only one episode. Fourteen percent of patients experienced transfusion reactions with bolus doses, but no reactions were observed in patients given CI. There were no severe reactions. All the reactions resolved following interruption of the infusion and administration of steroids. Premedication did not prevent reactions. In this series the median decrease in platelet count after bolus injection of pFVIII:C was -67 X 10(9) L(-1) compared with a median decrease of -2 x 109 L(-1) during the course of CI, thus, continuous infusion of pFVIII:C appears to have less effect on platelet count than bolus injection. An anamnestic response was associated with 77% of infusions. This high rate of anamnesis reflects patient selection, in that they were all known to have high-level high-responding FVIII inhibitors with cross-reactivity to pFVIII. After reconstitution, the pFVIII:C showed little loss in factor VIII activity in solution over a 24-h period. We conclude that pFVIII:C may be effectively administered by CI to patients with haemophilia A and high-responding FVIII inhibitors. CI is the probably the mode of administration of choice for intensive replacement therapy with pFVIII. PMID- 11851751 TI - Discontinuation of prophylactic therapy in severe haemophilia: incidence and effects on outcome. AB - A cohort study was performed to assess adherence to early prophylactic therapy and its effects on outcome in 49 patients with severe haemophilia born 1970-1980. Median age at start of prophylaxis was 5.5 years. The majority (69%) of patients interrupted prophylactic treatment one or more times of their own accord (median total interruption 2.2 years). Patients who discontinued prophylaxis at any point tended to have more arthropathy as measured by the Pettersson scale (median 8 points versus 4 points). One-third of these patients interrupted prophylaxis for longer periods and had permanently stopped taking prophylaxis at a mean age of 20.1 years (mean +/- SD duration 4.1 +/- 4 years) and consequently experienced 5.4 +/- 3.4) joint bleeds per year. This subgroup could be identified by a predictive score based on age at start of prophylaxis, weekly dose of prophylaxis, and joint bleed frequency on prophylaxis. In conclusion, while on prophylaxis, more than two-thirds of patients with severe haemophilia try to discontinue treatment, resulting in slightly more arthropathy. One-third of these patients permanently discontinue prophylaxis in adulthood, while maintaining a low number of joint bleeds. PMID- 11851752 TI - Complications of central venous catheters in patients with haemophilia and inhibitors. AB - We report our clinical experience with central venous catheters (CVCs) in 15 patients with haemophilia who, in total, had 34 catheters inserted. Eighteen devices were Hickman, six were Port-A-Cath and 10 were nontunnelled catheters (one Quinton, seven antecubital, one jugular and one subclavian vein access). All patients had factor VIII/IX inhibitors at the time of insertion. The mean age at operation was 8.8 years (range 16 months-39 years). Eight of the 15 patients (26/34 implanted catheters, 76%) presented some kind of complication. Pericatheter bleeding during the postoperative period affected a total of seven CVCs (7/34, 20%) in six patients, which required substitutive treatment for several days. Infection was reported in 15 of the CVCs (15/34, 44%), and four of these (4/15, 26%) had more than one episode, with a mean of 1.4 infection episodes per catheter (21/15). The infection rate was 0.2 infections per 1000 patient days or 0.1 per 1000 catheter days. Despite the usefulness of CVCs in haemophilic patients, the high incidence of complications requires careful assessment of the type of device as well as continuous surveillance. PMID- 11851753 TI - Total thrombin-activatable fibrinolysis inhibitor (TAFI) antigen and pro-TAFI in patients with haemophilia A. AB - Pro-thrombin-activatable fibrinolysis inhibitor (pro-TAFI), also known as TAFI, procarboxypeptidase U, or procarboxypeptidase B, is a relatively recently described plasma glycoprotein synthesized in the liver. It can be catalysed into its active form, TAFI (TAFIa, carboxypeptidase U or B) by a complex of thrombin/thrombomodulin. TAFI can potentially inhibit fibrinolysis by removing carboxyterminal lysine residues from partially degraded fibrin, decreasing plasminogen binding on the surface of fibrin, which thereby results in a decrease of the fibrinolytic activity. As TAFI represents a connection between coagulation and fibrinolysis, it can be expected that TAFI levels are altered in different thrombotic and haemorrhagic diseases, such as haemophilia A. Total TAFI antigen (including pro-TAFI, TAFI and the inactive form of TAFI [TAFIi]) and pro-TAFI were determined in 17 patients with haemophilia A. Thirteen healthy age-matched volunteers served as controls. No significant difference in levels of total TAFI antigen was observed between controls and patients with haemophilia, although it was slightly decreased in patients with haemophilia. Pro-TAFI was significantly reduced in haemophilia patients compared to controls (P=0.0113). TAFI antigen levels similar to controls have already been described in different clinical conditions, including haemophilia A. Decrease of pro-TAFI in haemophilia A can be an additional factor, together with decrease in thrombin generation, which induces impaired activation of pro-TAFI to TAFI, and could cause accelerated fibrinolysis. This supports the validity of usage of antifibrinolytics in the treatment of haemophilia A. In this paper we use new nomenclature for TAFI, and we believe that this recommended terminology for different forms of TAFI can simplify further standardization in TAFI investigation. PMID- 11851754 TI - Descriptive epidemiology of haemophilia in Maharashtra, India. AB - Secondary data on haemophilia cases in the state of Maharashtra, India were compiled and analysed. A total of 2192 haemophiliacs could be accounted from case papers and pedigrees, representing approximately 60% of the estimated number of cases in the state. Ratio of haemophilia A to haemophilia B was 4.2:1. Severe haemophiliacs constituted the majority of haemophilia A and B cases. About 70% of patients did not report any family history. Age distribution of cases suggested that the longevity of severe haemophiliacs was less than 30 years. Geographical distribution revealed clustering of cases around three established haemophilia clinics, with large parts of the state showing no cases. Although the cumulative number of cases showed an ascending trend, there was a 90% decrease in the number of cases registered between 1989 and 2000, suggesting that case recruitment from areas around the facilities had been achieved. Most cases showed delayed diagnosis, as the majority of severe haemophiliacs was being diagnosed at around 11-15 years of age. However, increasing awareness about haemophilia was reflected in a four-fold increase in referrals of severe cases over a 10-year period and increased registration of cases under 5 years of age. This study highlights that haemophilia services in India are still in their infancy, and increased awareness among healthcare providers as well as expansion of treatment facilities are still the primary areas for improvement of treatment. PMID- 11851755 TI - Hepatitis C viral clearance and antibody reactivity patterns in persons with haemophilia and other congenital bleeding disorders. AB - We studied hepatitis C virus (HCV) clearance and antibody reactivity patterns in a cohort of 100 haemophiliacs exposed to unsterilized blood products, of whom 25 were antiHCV negative and 75 were antiHCV positive [49 human immunodeficiency virus (HIV) negative and 26 HIV positive]. HCV RNA was measured by the 2.0 bDNA assay and an 'in-house' polymerase chain reaction assay. Antibody reactivity patterns were examined using a recombinant immunoblot assay (RIBA). Prior HCV infection was found in two (8%) of 25 antiHCV negative patients. HCV viraemia persisted in all 26 antiHCV+ patients who were coinfected with HIV. HCV RNA clearance was found in 12 (25%) of 49 antiHCV+, HIV- patients. Viral clearance was associated with younger current age (P < 0.01) and age at infection (P < 0.001), but not with duration of infection or with dose or frequency of clotting factor use. RIBA ratios reflecting an index of each patient's overall reactivity to four HCV epitopes were significantly lower in those with viral clearance (P < 0.0001). Over a period of 15 years, those with viral clearance demonstrated significant loss of reactivity to the NS3, NS4 and NS5 epitopes, while those with viral persistence demonstrated relatively stable reactivities to all epitopes. We conclude that spontaneous HCV RNA clearance in haemophiliacs is age-related and is unlikely to occur in those coinfected with HIV. The loss of antibody reactivity for some epitopes, especially c22 (core), may be a marker for the natural resolution of chronic HCV infection. PMID- 11851756 TI - Effects of HAART on hepatitis C, hepatitis G, and TT virus in multiply coinfected HIV-positive patients with haemophilia. AB - In multiply coinfected human immunodeficiency virus (HIV)-positive patients, we investigated the effects of high-activity antiretroviral therapy (HAART) using HIV protease inhibitors on three other viruses: hepatitis C virus (HCV), hepatitis G virus (HGV), and TT virus (TTV). Viral concentrations were measured serially by polymerase chain reaction methods in five patients with quadruple infection (HIV, HCV, HGV, and TTV) and in two patients with triple infection (HIV, HCV, and HGV) before and during HAART. In addition, CD4+ cell counts and serum alanine aminotransferase (ALT) levels were measured serially. Generally we observed no difference in serum HCV RNA, HGV RNA, or TTV DNA concentrations between samples obtained before and after initiation of HAART, whereas HIV RNA concentration decreased and CD4 counts increased in most patients. However, two patients had markedly decreased concentrations of HCV RNA and HGV RNA, respectively, more than 12 months after beginning HAART. Normalization of serum ALT levels was observed in a patient with decline of HCV RNA concentrations. No interactions were observed among these four viruses. HAART had no apparent direct effects on HCV, HGV, or TTV. Further studies will be required to elucidate whether the restoration of immune status through suppression of HIV replication by HAART may affect HCV or HGV RNA concentrations. PMID- 11851757 TI - Proprioception and isometric muscular strength in haemophilic subjects. AB - Haemophilia is characterized by intra-articular bleeding, often requiring immobilization, which may result in muscle atrophy and impaired proprioception. The aim of the study was to investigate differences in proprioceptive performance and isometric muscular strength of the lower limbs in haemophilic subjects compared with control subjects. Twelve subjects with severe haemophilia (11 haemophilia A; one haemophilia B) vs. 12 control subjects were matched for anthropometric data and tested for differences of proprioception (one-leg-stand, posturomed, angle-reproduction, and tuning fork tests) and isometric strength (leg press, knee extensor). The static proprioceptive performance of the haemophilic group, as measured by the one-leg-stand test (on hard or soft ground, with open or closed eyes; P < 0.05) was demonstrably impaired (by 41-363%). In contrast, the dynamic proprioceptive performance measured by the posturomed test did not show any difference between the groups. The local proprioceptive performance (angle-reproduction test) of the knee, (the most commonly affected joint in haemophiliacs) showed a trend to impaired function but was not distinctly different from that of controls. The quantitative sensory function (tuning fork) showed significant (P < 0.05) impairment of 9-10% in the haemophilic subjects. Additionally, the isometric muscular strength of the leg extensor was weaker (32-38%) in the haemophilic group when the limbs were tested individually as well as bilaterally (P < 0.05). In conclusion, the results suggest that global proprioceptive performance is impaired and that the isometric strength of the leg extensors is weaker in the haemophilic subjects. Therefore, specialized training for global proprioception would be helpful in order to compensate for proprioceptive deficits. This exercise regimen should also include safe strength-training for an optimal stabilization of the joints, but must be adapted to the individual needs and situations of the haemophilic subjects.p PMID- 11851758 TI - In vivo haemostatic effects of activated prothrombin complex concentrate and recombinant factor VIIa in a haemophilia A patient with inhibitors. PMID- 11851759 TI - Pseudotumour in von Willebrand disease. AB - Pseudotumour is a rare complication of haemophilia; it has previously been reported in patients with moderate or severe haemophilia and rarely in mild disease. We report a case of a proximal pseudotumour occurring in a 36-year-old patient with mild von Willebrand disease (vWD) who made a good recovery with conservative management. Surgery has been advocated as the optimal treatment for proximal pseudotumours due to the risk of continued bleeding and progression. However, in mild haemophilia or vWD, where the risk of spontaneous bleeds is low, conservative management may be an appropriate alternative. PMID- 11851760 TI - Haemophilic pseudotumour of the paranasal sinuses: management with radiotherapy and factor replacement therapy. AB - A case of pseudotumour of the paranasal sinuses occurring in a patient with haemophilia A is reported. There was a favourable response to combined treatment with radiation therapy and factor VIII replacement. PMID- 11851761 TI - Successful outcome of a cirrhotic patient with postoperative haematuria treated with a single high dose of recombinant factor VIIa. AB - Recombinant factor VIIa (rfVIIa) has been widely used for the treatment and prevention of bleeding episodes in haemophiliacs with high-titre inhibitors. High single doses are the treatment of choice for joint and muscle bleeds in those patients. There are only a few reports on the value of rfVIIa in cirrhotic patients with haemostatic impairment but this drug can consistently correct the prothrombin time in these individuals. We report a case of a good response to a single high dose of rfVIIa in a patient with advanced liver disease who suffered from severe refractory postoperative haematuria. PMID- 11851762 TI - Rapid prenatal diagnosis of haemophilia. PMID- 11851764 TI - Endoscopic ureteral incision using the holmium:YAG laser. AB - BACKGROUND: We reviewed the results of endoscopic ureteral incision for benign ureteral stricture, ureteropelvic junction obstruction and ureteroenteroanastomotic stricture using the holmium laser. METHODS: We carried out endoscopic ureteral incision using the holmium laser through an 8-Fr semirigid or 6.9-Fr flexible ureteroscope on 17 ureters in 15 patients. Balloon dilatation was not necessary before insertion of the ureteroscope. The stricture was incised with the holmium laser using a 200-365 microm fiber through the working channel of the ureteroscope. After completion of the incision, a 12-Fr double-J catheter was left for 6 weeks. Thereafter patients were followed by renal scan and/or ultrasound and excretory urography at 3-6 month intervals. RESULTS: The mean operative time was 65 min (18-135 min). The stricture resolved completely in 86.7% of cases at an average follow up of 20.5 months (11-32 months). CONCLUSIONS: The holmium laser endoscopic ureteral incision was associated with a good outcome in our series. We recommend this procedure to be employed initially because it is less invasive and has a favorable outcome. PMID- 11851766 TI - Prognosis of conservative therapy of advanced interstitial cystitis: experience of five cases. AB - BACKGROUND: It is generally believed that surgical procedures, particularly the more invasive ones, should be reserved for patients who fail to respond to conservative therapy by several established modalities. We studied whether conservative therapy was applicable to patients with interstitial cystitis (IC) in which an individual's awake bladder capacity decreases to 100 mL or less with vesicoureteral reflux (VUR). METHODS: Five cases that satisfied the criteria proposed by the National Institutes of Health were examined. The maximum awake bladder capacity was 50-100 mL (average, 74 mL). The bladder capacity under anesthesia was 70-300 mL (average, 199 mL). Only conservative treatments were carried out. RESULTS: The observation period ranged from 3 to 16 years with an average of 9 years. The awake bladder capacity ranged from 200 to 300 mL (average, 250 mL). The symptom index total score, which was previously 18.5, decreased to 5.5 and the problem index total score, which was previously 14.3, decreased to 3.5. Symptoms of bladder irritation improved. In four cases, VUR disappeared as the bladder capacity increased. Vesicoureteral reflux persisted in one case although its severity decreased and there was no fever. CONCLUSION: The patients treated with conservative therapy were able to maintain an awake bladder capacity that was sufficient for acceptable quality of life and their symptoms of bladder irritation improved. This suggests that we should regenerate the original remaining bladder using conservative therapy, even if IC is in a late stage. PMID- 11851765 TI - Clinical study of transitional cell carcinoma of the prostate associated with bladder transitional cell carcinoma. AB - BACKGROUND: Transitional cell carcinoma of the prostate in patients with bladder cancer appears to influence the prognosis and affects the decision about therapeutic modality. Therefore, it is important to characterize transitional cell carcinoma associated with bladder cancer. METHODS: From April 1980 to December 1998, 81 male patients underwent total cystoprostatectomies for transitional cell carcinoma of the bladder. The 81 cystoprostatectomy specimens were examined to clarify the characteristics of prostatic involvement by transitional cell carcinoma. The extent, origin, mode of spread and risk factor of prostatic involvement as well as the prognosis were investigated. In 13 of 15 patients with prostatic involvement the prostate was examined by sequential step sections. RESULTS: Prostatic involvement was observed in 15 of 81 patients (18.5%). Prostatic urethral involvement, invasion to prostatic duct/acinus, prostatic stromal invasion and extraprostatic extension and/or seminal vesicle involvement were recognized in 12 (80%), 14 (93.3%), six (40%), and five (33.3%) of the 15 patients, respectively. Twelve of the 15 patients (80%) with prostatic involvement had papillary or non-papillary tumors (i.e. carcinoma in situ) both in the prostatic urethra and prostatic duct. In 10 of these 12 patients (88.3%), there was contiguity between prostatic urethral and ductal tumors. Seven of the 23 patients (30.4%) with carcinoma in situ of the bladder showed prostatic involvement, which increased to 50% in the presence of carcinoma in situ of the trigone or bladder neck. CONCLUSIONS: Eighty per cent of the patients with prostatic involvement showed papillary or non-papillary tumors both in the prostatic urethra and prostatic duct. There was a high level of contiguity between both tumors. Patients with carcinoma in situ of the trigone or bladder neck revealed significantly higher incidence of prostatic involvement. PMID- 11851767 TI - Polytetrafluoroethylene patch sling for type 2 or type 3 stress urinary incontinence. AB - BACKGROUND: Surgical success rates and complications of anti-incontinence surgery were evaluated according to the type of stress incontinence and the type of surgery. METHODS: From 1989 to 1998, we treated 137 women for stress urinary incontinence with anti-incontinence surgery. Of the 137 patients, 110 had type 2 stress urinary incontinence and 27 had type 3 stress urinary incontinence. Of 110 patients with type 2 stress urinary incontinence, 57 underwent pubovaginal sling procedure and 53 were treated with the Gittes procedure. All of the patients with type 3 stress urinary incontinence underwent the pubovaginal sling procedure. RESULTS: Cure rates with the pubovaginal sling procedure were 82% in type 2 incontinence and 70% in type 3 incontinence. Cure rates with the Gittes procedure were 56%. The occurrence of de novo urge symptom and pelvic pain was low and bore no relation to either operative method or type of stress incontinence. CONCLUSIONS: The pubovaginal sling procedure using the polytetrafluoroethylene patch was effective for type 2 or 3 stress urinary incontinence. PMID- 11851768 TI - The pharmacokinetics of fosfestrol and diethylstilbestrol in chronic hemodialysis patients with prostate cancer. AB - BACKGROUND: Fosfestrol drip infusion therapy is an available endocrinotherapy for prostate cancer. But since there have been few reports of its use in chronic dialysis patients, the pharmacokinetics of fosfestrol in these patients remains unclear. We conducted fosfestrol drip infusion therapy as an induction therapy in chronic hemodialysis patients with prostate cancer. METHODS: Two male patients were included in this study. One was a 68-year-old man who had been in hemodialysis for 15.7 years and had stage B2 prostate cancer. The other was a 74 year-old man who had been in hemodialysis for 4.4 years and had stage C prostate cancer. A total of 250 mg of fosfestrol was dissolved in 250 mL of 5% glucose solution and administered by drip infusion. The drug was given subcutaneously during 14 consecutive days and a luteinizing hormone-releasing hormone agonist was injected on day 15. RESULTS: Serum fosfestrol levels increased rapidly after the drip infusion was started and remained at high levels during infusion, but fell quickly after the treatment ended. Diethylstilbestrol (DES) was also detected in blood after the infusion was started and its levels peaked when infusion ended. But on the next day, neither fosfestrol nor DES were detected in the blood of the patients. Moreover, neither fosfestrol nor DES was detected in the blood of the two patients before administering fosfestrol on day 15. Fosfestrol was quickly eliminated from the blood after hemodialysis was started, while DES remained in the blood during hemodialysis. The adverse reactions were mild hepatic dysfunction and gynecomastia. CONCLUSIONS: Fosfestrol drip infusion therapy appeared to be safe as an endocrinotherapy for prostate cancer in chronic hemodialysis patients. PMID- 11851769 TI - New concept parameters of RigiScan in differentiation of vascular erectile dysfunction: is it a useful test? AB - BACKGROUND: The aim of this study is to investigate the value of new nocturnal penile tumescence recording parameters, such as tumescence activity unit and rigidity activity unit values, total erection number and erection times, in differentiating between psychogenic erectile dysfunction and organic erectile dysfunction. We also aimed to determine the role of these parameters in differentiating arterial erectile dysfunction from veno-occlusive dysfunction. METHODS: Eighty-seven consecutive patients were allocated into three groups as psychogenic, arterial and venous erectile dysfunction after investigations. Nocturnal penile tumescence recording parameters between psychogenic and vascular erectile dysfunction and arterial and veno-occlusive dysfunction were compared. Mann-Whitney U-test, Pearson's chi2 test and correlation coefficient tests were used for statistical analysis. RESULTS: Depending on intracavernous injection, penile Doppler ultrasonography and cavernosometry tests, 37 patients (43%) had psychogenic impotence while 50 (57%) had organic pathologies. Of the 50 patients diagnosed with vascular impotence, 29 (48%) had arterial failure and 21 (42%) had veno-occlusive dysfunction. Nocturnal penile tumescence recording revealed psychogenic erectile dysfunction in 34 patients (39%) and vascular erectile dysfunction in 53 patients (61%). Nocturnal penile tumescence recording has been regarded as the gold standard and, in our series, it showed 90.6% sensitivity and 88.2% specificity in differentiating the cause of erectile dysfunction. Values of rigidity activity unit and tumescence activity unit were significantly higher in patients with psychogenic impotence (P < 0.001), when compared with vascular impotence. In patients with a vascular cause, no difference was found between arterial failure and veno-occlusive dysfunction with regard to tip tumescence activity unit, base tumescence activity unit, tip rigidity activity unit, base rigidity activity unit and erection time (P > 0.001). However, patients with arterial failure had less erection than patients with veno-occlusive dysfunction (P < 0.001). CONCLUSION: New recording parameters of nocturnal penile tumescence can differentiate organic and psychogenic erectile dysfunction more precisely. However, these recording parameters cannot distinguish subgroups with a vascular cause of erectile dysfunction. PMID- 11851770 TI - Laparoscopic diagnosis and treatment of nonpalpable testis. AB - OBJECTIVE: Laparoscopy has become one of the important diagnostic modalities of nonpalpable testis and has been developed and applied in the treatment of this disease. In the present study, we investigated the usefulness of laparoscopy in the diagnosis and treatment of nonpalpable testis. METHODS: Laparoscopy was carried out under general anesthesia on 21 patients (23 testes) from October 1991 to October 1999. If the internal spermatic vessels and vas deferens made their way into the internal inguinal ring, the inguinal canal was dissected with an incision in the inguinal region to look for the testis. Patients with intra abdominal testis underwent laparoscopic orchiopexy or orchiectomy. If the internal spermatic vessels terminated with a blind end intraperitoneally, making it impossible to identify the testis, the case was judged to be vanishing testis and the operation was finalized without any further examination. RESULTS: In eight of 23 testes (35%), the internal spermatic vessels and vas deferens made their way into the internal inguinal ring. The inguinal region was examined in all the eight testes. Orchiopexy was carried out on two testes and orchiectomy was carried out on six testes. An intra-abdominal testis was detected in eight of 23 testes (35%). Laparoscopic orchiopexy was carried out on seven testes. One stage orchiopexy was carried out on two of the seven testes and two-stage Fowler Stephens orchiopexy was carried out on five of the seven testes. Orchiectomy was carried out on the remaining testis. Blocking or lack of the internal spermatic vessels and vas deferens was seen in seven of the 23 testes (30%) and this condition was diagnosed as vanishing testis. CONCLUSION: Laparoscopy for nonpalpable testis is considered to be the most effective technique for diagnosing the presence or absence of the testis and the location of the testis. PMID- 11851771 TI - Proliferative activity, angiogenesis and nuclear morphometry in renal cell carcinoma. AB - BACKGROUND: Prognostic parameters other than tumor stage and grade are essential for renal cell carcinoma (RCC) patients. This study was undertaken to determine the usefulness of cellular proliferation, angiogenesis and nuclear morphometry in predicting the biological aggressiveness of RCC. METHODS: Surgical specimens of 70 patients with RCC were investigated by conventional histology, Ki-67 immunostaining and stereological assessment of angiogenesis and mean nuclear volume. RESULTS: There was no difference in disease-specific survival with respect to sex, age and histopathological type (except sarcomatoid and other types). The survival was significantly lower and the chance of metastases was higher in the group with higher proliferative activity (P=0.007). There was no relation between angiogenesis, mean nuclear volume, stage and survival. There was a significant relation between both Fuhrman and WHO grades, tumor stage and survival. Histopathological type, grade, angiogenesis and mean nuclear volume failed to predict recurrences and/or metastases. In multivariate analysis, only TNM stage and proliferative activity were found to be independent prognostic factors. CONCLUSIONS: In addition to tumor grade and stage, proliferative activity of a given RCC may have the potential to identify patients with an impaired prognosis. PMID- 11851772 TI - Sarcomatoid transitional cell carcinoma originating from a duplicated renal pelvis. AB - A case of sarcomatoid transitional cell carcinoma of the renal pelvis is reported. It was distinguished from carcinosarcoma by immunohistochemical study. The tumor was difficult to distinguish from a renal parenchymal tumor in imaging studies because it originated from a duplicated renal pelvis. PMID- 11851773 TI - A stone developed within the dilated intravesical ureter following ureteroscopy. AB - A 58-year-old man presented with a stone within the dilated intravesical ureter, which was probably attributable to a previous ureteroscopy. Transurethral incision of the right intravesical ureter and lithotripsy were carried out without subsequent urinary tract impairment. Although some complications resulting from ureteroscopy, such as ureteral stricture, ureteral perforation and vesicoureteral reflux, have been reported, this complication is considered to be very rare. PMID- 11851774 TI - Testis sparing surgery for the treatment of a sequential bilateral testicular germ cell tumor. AB - Standard therapy of sequential bilateral testis cancer is generally considered to be orchiectomy. We present a case of sequential bilateral testicular germ cell tumor treated with testis sparing surgery. The patient was disease free 50 months after surgery without local recurrence or distant metastases. Testis sparing surgery provides a better quality of life and may be considered a safe, feasible alternative in the treatment of carefully selected patients with bilateral testicular germ cell tumor. PMID- 11851775 TI - Flexible video cystoscope with built-in high-frequency cauterizing element for transurethral resection of bladder tumor. AB - The major advantage of the flexible video cystoscope is that a digital signal can be obtained while high frequency cauterization is carried out. Cauterization while observing a digital signal picture was not possible before this new model was developed. We decided to use this new cystoscope to resect a bladder tumor and coagulate the bleeding because the patient could tolerate only local anesthesia due to severe heart disease complications. We successfully treated the patient with this technique and no complications were noted. This new flexible video cystoscope was found to be safe for resecting bladder tumor under local anesthesia. PMID- 11851776 TI - A case of pelvic malignant paraganglioma. AB - We report a rare case of pelvic malignant paraganglioma that was treated with surgery, combination chemotherapy and radiation. A 47-year-old man was diagnosed with pelvic malignant paraganglioma that had metastasised to the thoracic vertebrae. The pelvic mass, which was 6 cm in size, was on the posterior side of the bladder and had invaded the prostate, seminal vesicle and bladder neck. We resected the intrapelvic tumor and lymph nodes using cystoprostatectomy. Metastases to bilateral obturator lymph nodes and the right internal iliac lymph node were shown by pathology. Adjuvant therapies included six courses of the combination chemotherapy (cyclophosphamide, vincristine and dacarbazine), and 12 courses of VP-16 therapy. Radiation therapy was done for metastasis of the thoracic vertebrae. Local recurrence, progression of bone metastasis and new metastasis have not been detected since these treatments. The patient has been clinically stable during 20 months of follow-up. Chemotherapy of cyclophosphamide, vincristine and dacarbazine and VP-16 with radiation appears to be effective in treating advanced malignant paraganglioma. PMID- 11851777 TI - Embryonal carcinoma of the testis associated with prostate cancer in a 72-year old man. AB - A 72-year-old Japanese man presented with a painless swollen left scrotal mass with elevated levels of serum alpha-fetoprotein and prostate specific antigen. The patient underwent high orchiectomy under diagnosis and a final pathological examination revealed embryonal carcinoma of the left testis. A systematic needle prostate biopsy under guidance of transrectal ultrasound revealed prostate cancer (Gleason score, 8) on the left lobe (T2aN0M0). Systemic chemotherapy was given for retroperitoneal lymph node metastasis of testicular cancer and hormonal therapy (LH-RH analog) was given for prostate cancer. The patient was well with no evidence of metastasis from the testicular cancer or prostate cancer and with no elevation of serum alpha-fetoprotein or prostate specific antigen 26 months after the orchiectomy. PMID- 11851780 TI - A qualitative study of shift handover practice and function from a socio technical perspective. AB - BACKGROUND: A qualitative study of shift handover practice and function from a socio-technical perspective Background. Shift handover plays a pivotal role in the continuity of patient care in 24-hour nursing contexts. The critical nature of this communication system is recognized within the literature and by the nursing profession; however, there are few in-depth studies. The rationale for this study is to gain a better understanding of handover practices and functions and their implications for effectiveness. METHOD: Handover systems on two very different paediatric wards were selected as case studies. In each case, 20 handovers were observed and audio-taped and 12 individual and two-group interviews with nursing staff about handover were also conducted. Analysis involved categorizing the data and characterizing handover practices and functions using an inductive approach to generate qualitative themes. The ethics committees of the hospital and the university approved the research. All involved were fully informed about the study, with confidentiality maintained throughout. RESULTS: Handover practices are distributed over time, socially among the staff and technologically through a range of artifacts, while the system also accomplishes informational, social and educational functions. Handover effectiveness is characterized by flexibility in managing competing demands and tensions, such as maintaining confidentiality while practising family centred care. There are limitations in how far the findings can be generalized to other nursing contexts, and the possible effects of the researcher's presence are also recognized. CONCLUSIONS: Handover is a complex system based on several sound socio-technical principles and the value of this nurse-to-nurse communication should be acknowledged. The multiple functions highlight the knowledge and expertise currently hidden within handover, which could be promoted in terms of nursing professionalism. PMID- 11851781 TI - Using ICIDH-2 in the classification of nursing diagnoses: results from two pilot studies. AB - BACKGROUND: Using ICIDH-2 in the classification of nursing diagnoses: results from two pilot studies Background. As an International Classification of Functioning and Disability, the ICIDH-2 is potentially relevant to the work of all health care professionals. Nurses, however, had little involvement in the development of this classification and have no tradition of using it. AIM: Two pilot studies were designed to explore the fit between ICIDH-2 and the nursing domain. METHODS: In the first study, experts (n=2) and nurses (n=9) were asked to identify and classify patient problems that they observed on a video taped case. In the second study, problem statements from nursing diagnoses (n=199) that had been retrieved from patient files were classified using ICIDH-2. Each problem statement was classified by a panel of three individuals. RESULTS AND CONCLUSION: Results from the two studies suggest that the ICIDH-2 is certainly relevant to the nursing discipline and allows a large majority of nursing diagnoses to be classified. It is recommended that nurses take an active role in the ongoing development of the ICIDH in order to further improve its usefulness to the nursing profession. LIMITATIONS: The studies were small-scale pilots performed in a single medical centre. Preliminary insight resulted from these pilots, but more research is needed. PMID- 11851782 TI - Maintaining privacy on post-natal wards: a study in five European countries. AB - AIM: Maintaining privacy on post-natal wards: a study in five European countries Aim. To describe how mothers' privacy is maintained on post-natal wards in five European countries: Finland, Spain, Greece, Germany and Scotland [United Kingdom (UK)] BACKGROUND: Privacy is an important concept in nursing and nursing ethics. Empirical studies in this field, however, are few. In this study, privacy is defined as comprising both physical and social-informational dimensions and both the perceptions of mothers and professional were investigated. METHODS: The data were collected by a questionnaire from mothers (n=1192) and midwife/nursing staff (n=952). RESULTS: The results revealed differences both between the different countries and between patients' and professionals' views. Mothers in Finland, Germany and Scotland felt their privacy was maintained better than mothers in Greece and Spain. The differences in staff perceptions are less clear-cut. Different background factors emerge as statistically significant in different countries. CONCLUSION: Empirical studies in privacy are important and give ideas about the problems in nursing practice. Future analyses should look also at other areas of nursing. PMID- 11851783 TI - Minimizing side-effects: the clinical impact of nurse-administered 'side-effect' checklists. AB - BACKGROUND: For those with chronic illness, the adverse effects of medication are important causes of morbidity and distress which may not always receive due attention. Guidelines and checklists may be one strategy to focus professionals' attention on long-term problems. Therefore, client-centred 'side-effect' evaluation checklists were developed to be administered and actioned by nurses. AIM: The purpose of this study was to explore the clinical impact of these checklists on long-term users of antipsychotic medication. METHODS: This study was undertaken with clients with enduring mental illness, in Community Mental Health Teams. In the first phase, 40 nurse-client interactions were observed. Following introduction of the evaluation checklists, 20 nurse-client interactions were observed with the checklists and 20 nurse-client interactions were observed for comparison. In addition, the views of professionals, service users and user groups on the value of the checklists were sought. FINDINGS: Amongst the 20 clients in the intervention group, the checklists highlighted several problems, two of which were urgent. In the intervention group, the mean number of problems actioned per client increased from 0.35 (range=0-4) with no checklists to 3 (range=0-6) with the checklists. The majority (51 of 59) of actions taken to alleviate adverse effects of medication concerned physical health problems. Nurses offered appropriate advice or encouraged clients to contact the relevant agencies. No such changes were observed in the comparator group. There were no differences between groups in the number of referrals to prescribers. IMPLICATIONS: The usefulness of the evaluation checklists for detecting unattended problems, in conjunction with the responses of service users, suggests that it may be clinically effective to establish procedures to systematically monitor people with serious mental illness for adverse drug reactions (ADRs). However, larger studies are needed to confirm this. PMID- 11851784 TI - Barriers and bridges to care: voices of homeless female adolescent youth in Seattle, Washington, USA. AB - AIM: The purpose of this study was to conduct an assessment of reproductive health-seeking behaviours, sources of advice, and access to care issues among a sample of clinic-based homeless adolescent women. BACKGROUND: Adolescent women are among the most vulnerable and medically underserved subgroups within the homeless population in the United States. Homeless youth are rarely invited to participate in research aimed at improving their access to appropriate health care. Also, the culture in which they live and the personal experience of being homeless are often not addressed. STUDY METHOD: The research was descriptive, using focus groups and individual interviews with a purposeful sample of 20 female youth, aged 14-23 years. FINDINGS: The women said that they seek health advice from other women, including their mothers even while they are homeless. They reported first trying self-care interventions, and going to clinics when self-care actions no longer worked. They stated that the main barriers to health care were lack of insurance, confusion over consent, transportation problems, lack of respect (from providers) for their own self-knowledge, and judgementalism from providers. CONCLUSION: Using the concept of cultural competency, the results provide insights into how to improve communication and health care services for these women. RECOMMENDATIONS: Health care providers need to recognize and appreciate the lifestyle, beliefs, and adaptive attitudes of homeless youth, rather than labelling them as 'deviant'. All personnel who interact with and on behalf of homeless youth must be adequately trained in general knowledge regarding the health of homeless youth as well as in an understanding of the role that culture plays in their health-seeking behaviours. PMID- 11851785 TI - The lived experience of waiting-to-know: Ebola at Mbarara, Uganda--hoping for life, anticipating death. AB - PURPOSE PF THE STUDY: The purpose of the study was to describe the phenomenon of 'waiting to know'. It is a phenomenon uniquely experienced by persons who had been exposed to patients with Ebola Hemorrhagic Fever (Ebola) but who have not yet exhibited signs and symptoms of the disease. RESEARCH METHOD/ANALYSIS: The phenomenological human science approach was used using the four life worlds as guides for reflection. These are spatiality, corporeality, temporality, and relationality. PARTICIPANTS: Seven health care personnel were selected through professional networking. They were preferred because of their exposure to patients with Ebola during the epidemic at Mbarara, Uganda. RESULTS/FINDINGS: Written descriptions of the experience of 'waiting to know' whether exposure to patients with Ebola causes these participants to be infected, were obtained and analysed. Through immersion with the written descriptions, the following themes emerged: helplessness in anticipation and fear of dying or premature death; agonizing and languishing over losing relatives, friends, and loved ones; trusting no one, and; helplessness and hopelessness with the persistence of time. PMID- 11851786 TI - Self-reported health and glycaemic control in Tanzanian and Swedish diabetic patients. AB - AIMS OF THE STUDY: To investigate self-reported health in adult Tanzanian and Swedish diabetic patients in relation to the general population in the two countries and to investigate whether diabetic patients with poor glycaemic control also rated their self-reported health to be impaired. DESIGN/METHODS: The study design was cross-sectional and comparative. One hundred and fifty Tanzanian patients were age- and gender-matched with Swedish diabetic patients. Self reported health was measured using the generic SF-36 health questionnaire, measuring eight different health domains. Glycaemic control was measured by testing glycosylated haemoglobin (HbA1c). RESULTS: The main results were that Tanzanian diabetic patients had poorer health in comparison with general Tanzanian population living in the same geographical area. In contrast, diabetic patients in Sweden did not markedly differ from the Swedish general population. Furthermore, Tanzanian patients had poorer glycaemic control. In both countries poor glycaemic control did not associate with impaired self-reported health, with one exception. Tanzanian patients with unsatisfactory or poor glycaemic control had significantly poorer reported health in the mental health domain. CONCLUSIONS: The results indicated that patients' health should be assessed using a specific health measure in addition to general medical measures. PMID- 11851787 TI - Preferences for female and male nurses: the role of age, gender and previous experience --year 2000 compared with 1984. AB - BACKGROUND: The situations in which patients prefer a male or female nurse are not clear, as the literature reports differing relationships between gender and preference. Furthermore, most research has focused on female preferences. METHODS: This questionnaire study investigated the preferences of both males and females for a male or female nurse, depending on four clinical situations. The role of age and previous experience with a male nurse was also taken into account. Gender, age and experience were further examined in relation to attitudes towards professional nursing practice. To determine whether preferences and attitudes have changed over time, the study was replicated in 2000, having been first conducted in 1984. Data were collected from the same general practice waiting room on each occasion. RESULTS: Consistent with previous research, the degree of intimacy in a clinical situation was found to be predictive of same gender preferences. Younger females may prefer female nurses more than older females. Experience with male nurses was limited in both samples, but was not predictive of preferences or attitudes. CONCLUSIONS: Generally speaking, results from 1984 and 2000 were not significantly different. As a quantitative study, this research contributes to an understanding of patient preferences. However, in depth, qualitative research is recommended to understand better the reasons for preferences and attitudes, for both male and female patients. PMID- 11851788 TI - The impact of foot massage and guided relaxation following cardiac surgery: a randomized controlled trial. AB - BACKGROUND: Because of the widely presumed association between heart disease and psychological wellbeing, the use of so-called 'complementary' therapies as adjuncts to conventional treatment modalities have been the subject of considerable debate. The present study arose from an attempt to identify a safe and effective therapeutic intervention to promote wellbe ing, which could be practicably delivered by nurses to patients in the postoperative recovery period following coronary artery bypass graft (CABG) surgery. Aim. To investigate the impact of foot massage and guided relaxation on the wellbeing of patients who had undergone CABG surgery. METHOD: Twenty-five subjects were randomly assigned to either a control or one of two intervention groups. Psychological and physical variables were measured immediately before and after the intervention. A discharge questionnaire was also administered. RESULTS: No significant differences between physiological parameters were found. There was a significant effect of the intervention on the calm scores (ANOVA, P=0.014). Dunnett's multiple comparison showed that this was attributable to increased calm among the massage group. Although not significant the guided relaxation group also reported substantially higher levels of calm than control. There was a clear (nonsignificant) trend across all psychological variables for both foot massage and, to a lesser extent, guided relaxation to improve psychological wellbeing. Both interventions were well received by the subjects. CONCLUSIONS: These interventions appear to be effective, noninvasive techniques for promoting psychological wellbeing in this patient group. Further investigation is indicated. PMID- 11851790 TI - Response to: Critical review of R.R. Parse's "The human becoming school of thought. A perspective for nurses and other health professionals.". PMID- 11851789 TI - Infant feeding responsiveness: a conceptual analysis. AB - BACKGROUND: The phenomenon of infant responsiveness to the caregiver during feeding interactions has been of interest to researchers for many years, because of its associated implications for child growth and development. Although many studies have examined feeding responsiveness in caregiver-infant dyads, it is infrequently and inconsistently defined in the literature. SPECIFIC AIMS: The purpose of this paper is to clarify the concept of infant feeding responsiveness through the provision of a working definition of the concept for further study and usage. METHODS: Medline, CINAHL, and PschInfo databases from the year 1970 to the present were searched for English articles containing the keywords 'infant', 'feeding', 'responsiveness', 'synchrony', and/or 'interaction'. Articles were selected for inclusion according to whether or not the phenomenon of infant feeding responsiveness was defined or assessed. Walker and Avant's (1995) method for concept analysis was employed for the development of defining attributes, case examples, antecedents, and consequences for further clarification of the concept of infant feeding responsiveness. FINDINGS: Infant feeding responsiveness was defined as the manifestation of physiologically influenced visual, expressive, vocal and motor reactive behaviours expressed by an infant in reaction to a caregiver's feeding attempts, indicating a readiness to feed. CONCLUSIONS: Implications for nursing are explored as well as the need for refinement of measures of this concept. PMID- 11851792 TI - Efficacy of the Shinki bioclean room for preventing infection in neutropenic patients. AB - AIM OF THE STUDY: To investigate the effectiveness of a new type of bioclean room named Shinki bioclean room (SBCR) for the prevention of infection during neutropenia after intensive chemotherapy in comparison with a standard laminar air flow room (LAFR). BACKGROUND: Recently, a new industrial technology, wherein a dust-free and aseptic environment is created by circulating air containing nanometre order ultra fine water droplets with abundant negative air ions, has been developed in Japan. METHODS: The air cleanliness of SBCR was examined by measuring airborne particles and microorganisms. Bacteriological samples for environment culture were taken by means of exposed settle-plates. In addition, the frequency of pneumonia and fever higher than 38 degrees C were examined in 34 patients with acute leukaemia who received intensive chemotherapy in SBCR or LAFR. RESULTS: The number of airborne particles (> or = 0.5 microm) was 70 particles/ft3, and that of airborne microorganisms was 0.0 colony forming unit/ft3 in SBCR, and neither bacteria nor fungi were detected. The numbers of colonies of bacteria and fungi on air settle-plates were fewer in the SBCR than in the LAFR regardless of the presence of patients or the nurse entering. The frequency of pneumonia during chemotherapy for acute leukaemia was lower in the SBCR group (0%, 0/19 cases) than in the LAFR group (27%, 4/15 cases) (P=0.0294) and the frequency of fever higher than 38 degrees C also tended to be lower in the SBCR group (53%, 10/19 cases) than in the LAFR group (80%, 12/15 cases) (P=0.0973). CONCLUSION: The SBCR is equal or superior to LAFR in preventing infection during neutropenia. Other advantages for SBCR are a low level of noise (40 dB), easy control of temperature and humidity, and efficient removal of odour. In addition to the quiet and comfortable atmosphere, expected favourable effects of negative air ions may give higher quality of life for patients in SBCR than those in LAFR. Further studies will be needed to examine the safety, benefits and effects of the negative ion exposure. PMID- 11851793 TI - Psychosocial consequences of falling: the perspective of older Hong Kong Chinese who had experienced recent falls. AB - AIM OF THE STUDY: The study's aim was to explore the psychosocial consequences of falling with a group of older Chinese who had recently fallen. BACKGROUND: Older people fall more frequently. Thus, the consequences of these falls and their influence on health outcomes need to be determined. One important outcome, namely the psychosocial consequence of falling, has not been extensively studied. As a result, this study explored the psychosocial consequences of falling with a group of older Chinese who had recently experienced a fall. RESEARCH APPROACH: An explorative qualitative approach with semi-structured interviews was used in this study. SAMPLE: Twenty informants, with recent fall experiences either in the community or hospital setting, were interviewed in two elder care wards in an acute care hospital. FINDINGS: Three major categories of psychosocial consequences of falling emerged from the interview data: powerlessness, fear and seeking care. Powerlessness was also exemplified in three subcategories: lack of control, self-comforting and lack of emotion. Informants perceived falls as unpredictable and not preventable, expressing fears that falling could result in dependence on others and becoming a care burden. The interview data also showed that there is a need by older Chinese to seek care and advice from relatives and health care professionals. CONCLUSIONS: Findings from this study have provided insights into the psychosocial consequences of falling for older Chinese. These insights suggest nursing interventions should promote a sense of mastery in prevention of falls, facilitate supportive social interactions with relatives and give empathetic responses to those who have fallen. PMID- 11851794 TI - The requirement for informed consent prior to nursing care procedures. AB - AIM OF THE PAPER: The aim of this paper is to examine the extent to which there is a requirement to obtain informed consent prior to nursing care procedures. RATIONALE: The requirement for nurses to obtain consent prior to nursing care procedures is addressed in various nursing policy documents. It is important that nurses understand the legal and ethical rationale behind the principles of informed consent so that the principles are applied appropriately to the particular context of nursing care. ARGUMENT: The ethical and legal rationale behind the concept of informed consent and its relevance to nursing practice are examined. In this paper, it is argued that the function of informed consent is to protect patient autonomy and to promote meaningful decision-making. Given the potential for nursing care procedures to infringe patient autonomy, consent is clearly a relevant concept in nursing. Furthermore, in law, any touching without consent is a potential battery. Informed consent is often associated as a rigid procedure, only relevant to surgical or research procedures. Consent should be obtained prior to nursing care procedures whenever patient autonomy is at stake. However, information-giving should be determined by the needs of the patient and approached in such a way as to facilitate meaningful decision-making. Given the individual nature of infringements to patient autonomy, it is difficult to predetermine all those care procedures that require consent; any list of procedures would fail to be comprehensive. CONCLUSIONS: The principles of informed consent should underpin our approach to nursing care procedures, which should not be mechanistic but determined by the needs of individual patients. PMID- 11851795 TI - What is the evidence on evidence-based nursing? An epistemological concern. AB - AIM: This paper adopts an epistemological perspective in order to support the assertion that there is very little evidence to support the existence of evidence based nursing as a distinct construct or process. RATIONALE: This analysis of concept meaning is based on the theory of symbolic interactionism. By adopting this theory the commonalities and inconsistencies in the use of evidence-based symbolism by nurses and other health care professionals can be used to evaluate the perceived meaning of the term. METHODS: A frequency analysis of relevant key words in one publications data-base demonstrates the increasing use of evidence based terminology and its euphemistic status visa vi research. An epistemological analysis is conducted on a sample of concept clarification statements taken from the popular literature, defining the nature of 'evidence' and 'evidence-based practice'. FINDINGS: The results of the frequency analysis show that the keywords 'evidence-based medicine' revealed 5612 papers, evidence-based practice (EBP) 432 papers, evidence-based nursing 47 papers, evidence-based health care 60 papers, and evidence-based decision making 43. Almost all of these papers have been published since 1995 and the earliest use of the symbol 'evidence-based' is 1992. There is also an increase in papers adopting 'evidence-based' symbolism along with a commensurate decrease in the use of the term 'research' in the nursing context. CONCLUSION: 'Evidence-based practice' is commonly a euphemism for information management, clinical judgement, professional practice development or managed care. There is insufficient evidence to demonstrate that evidence-based nursing is a single construct or process that can be distinguished from its concomitants. The term adds little more to the existing long standing traditions of quality assurance and research-based practice. Nurses must avoid the inefficiency brought about by the 'intense enthusiasm followed by sad disenchantment' that has been associated with other attempts to introduce innovation in health care delivery (Fienstein & Horowitz 1997). PMID- 11851796 TI - Developing culturally competent researchers. AB - BACKGROUND: Whilst we live in multicultural societies most health researchers tend to take the cultural perspective of the majority ethnic group at the expense of the perspective of minority ethnic groups. AIM: This paper discusses the need for the development of culturally competent health researchers in all areas of research and proposes a model for the achievement of this. DESIGN: A snapshot review of research textbooks used in nursing curricula was conducted to identify whether culturally competent research was being promoted. RESULTS: The review found that whilst a few textbooks touched on ethnicity, race and culture, none of them addressed the issue of cultural competence. Subsequently the authors adapted their existing model of culturally competent health care practice, and in this paper they propose it as a model for the development of culturally competent researchers. DISCUSSION: The model put forward by the authors consists of four concepts: cultural awareness, cultural knowledge, cultural sensitivity and cultural competence. A culturally competent researcher is one who is able to apply the related skills and knowledge in project design, data collection, analysis, report writing and dissemination. Furthermore, the authors identify two layers of cultural competence, those of culture-generic (knowledge and skills that are applicable across ethnic groups) and culture-specific competence (knowledge and skills that relate to a particular ethnic group). The relationship between these two layers is a dynamic and spiralling process as illustrated by the model. CONCLUSION: Current health policy in many developed countries focuses on inequalities of health and managing diversity, including ethnicity. Thus the authors conclude that the development of culturally competent researchers will lead to both valid research and culturally competent practice by health care professionals. PMID- 11851797 TI - Stages in coming to terms with reflection: student advanced nurse practitioners' perceptions of their reflective journeys. AB - AIM OF STUDY: To explore student Advanced Nurse Practitioners' (ANPs) experiences of reflection. BACKGROUND: The development of student ANPs' reflective abilities as part of the implementation of a Master's Course in Advanced Nursing Practice at Wolverhampton University provided the background to the study. Developing reflective abilities was seen as an essential component of ANP development. To facilitate this process a preentry degree level reflective practice module and a reflective component for the level 4 (Master's level) practice module were devised. RATIONALE: This was a new educational initiative and this study provided the opportunity to explore students' experiences of it following completion of the reflective modules. DESIGN/METHODS: A qualitative methodology was used. Fourteen ANP students were interviewed and their reflective learning contracts were analysed. Colaizzi's (1978) seven-stage model guided data analysis. FINDINGS: The findings were categorized under the headings of: (i) reflection as a transforming process, (ii) reflective journeys, (iii) worldviews. This paper presents an analysis of the second category reflective journeys. It identifies a number of stages in the development of students' reflective abilities. CONCLUSIONS: The conclusions have implications for those involved in reflective curriculum development, and for those seeking to develop reflective abilities in students. PMID- 11851798 TI - Laddered questions and qualitative data research interviews. AB - BACKGROUND: Nurse researchers frequently make use of open interviews in their studies, hoping to understand the private world of respondents, and in particular the ways in which thought and behaviour are connected to each other. Unfortunately, there is scant guidance on how to translate such goals into the specifics of interview technique, particularly the use of probing questions. The success of such interviews rests on working ethically and understanding the respondents needs during interview. AIMS: This paper explains a technique designed to direct the use of probing in interviews, and argues that its selective use might provide richer data than other ad hoc approaches employed by neophyte researchers. CONCLUSION: It is argued that the technique increases the awareness of the researcher to ethical concerns and enables her or him to respond more sensitively. Laddered question technique is evaluated with reference to my own research into the negotiation of student support amongst nurse distance learners. PMID- 11851799 TI - Phenomenology as a method to investigate the experience lived: a perspective from Husserl and Merleau Ponty's thought. AB - AIM: By taking nursing as a human relationships activity, in spite of its strong technical--scientific features, this article reflects on the phenomenological method as one of the ways to develop an investigation and acquire knowledge of the topic. RATIONALE: Based on Husserl's phenomenology, which is opposed to the way of doing science based on the laws that regulate the physics and mathematics, the article introduces Merleau Ponty's existential phenomenology as the theoretical foundation for the method it proposes. My existential conceptions- people as historic beings inserted in a world over which they act but which, in its turn, determines them; the human perception as reference for our way of being in the world; the space-time structure of perception--these are the key concepts that have led to the elaboration of an approach to phenomenological research. PROPOSAL OF A METHODOLOGY: Steps are proposed for such an approach, namely phenomenological description, reduction and analysis. These lead to the building up of ideographic and nomothetic analyses, thus unveiling and describing general truths about the phenomenon studied. Finally, the possibilities for applying the methodology to nursing research are discussed, illustrated by my research into student nurses' perspectives on working on an isolation ward. PMID- 11851800 TI - Perceptions of sex-role stereotypes, self-concept, and nursing role ideal in Chinese nursing students. AB - AIM: This study examined the relationship between sex-role stereotypes, self concept and the requisite personality characteristics of an ideal nurse in a cohort of Hong Kong nursing students. METHODS: To rate these concepts a measure of eight comprehensive dimensions of personality perception was administered to 177 nursing students, studying on preregistration and postregistration programs at a Hong Kong tertiary institution. Both male and female nursing students perceived an ideal nurse to possess a profile of traits including being high on the dimensions of emotional stability, application, intellect, helpfulness and restraint. RESULTS: No significant difference between the self-ratings of the male and female students was found, indicating that male students had undergone a highly self-selective process when choosing nursing education under the influence of Chinese cultural stereotypical attitudes towards nursing. A typical Chinese nurse was rated as similar to the typical female in Chinese society by both male and female nursing students. A typical Chinese nurse was rated relatively low on the masculine dimensions of openness, extroversion and assertiveness. The self ratings of male nursing students more closely approximated the ideal nurse than did the self-ratings of female nursing students. CONCLUSION: The conclusions highlight implications for the recruitment and education of both male and female nursing students in Hong Kong society. PMID- 11851801 TI - Differences in urinary stress hormones in male and female nurses at different ages. AB - AIM: The aim of this study was to assess the levels of urinary stress hormones in male and female nurses and to determine their contribution to sex-determined health outcomes. RATIONALE/BACKGROUND: While the use of questionnaires have shown that there is a high degree of stress in health care workers, these results are inconclusive. In this study a more objective approach was used by assessing the levels of urinary stress hormones, adrenaline, noradrenaline and cortisol. In premenopausal women oestrogen may attenuate sympathetic nervous system activity and the secretion of adrenaline and cortisol and therefore lower the stress response and incidence of stress-related illnesses. METHODS: Three hundred and fifteen nurses of both sexes were separated into two age groups, 20-40 and 45-60 years, and for postmenopausal women into those taking or not taking hormone replacement therapy (HRT). Early morning urine sample were collected and stress hormones determined by high performance liquid chromatography (HPLC). RESULTS: Urinary cortisol, adrenaline and noradrenaline levels were all increased with age in both sexes, with a greater difference in the younger age group compared with the older group. These hormones were lower in premenopausal women compared with male nurses of similar age. The difference between the sexes in the high age group was less compared with the lower one. Postmenopausal women on HRT had lower levels of these hormones. CONCLUSION: The age-related changes were surprising but may be because of higher stress levels in the older groups of both sexes. However, the gender difference supports the view that oestrogen reduces sympathetic activity and the secretion of cortisol and adrenaline. HRT reduces the stress response and therefore may have additional benefits by reducing the level of stress-related illnesses. These results were surprising but may suggest that oestrogen reduces the stress response in women and therefore offers protection against stress-related disorders. PMID- 11851802 TI - Enteric viruses in the aquatic environment. PMID- 11851803 TI - Evaluation of potential biocontrol rhizobacteria from different host plants of Verticillium dahliae Kleb. AB - AIMS: A screening approach was developed to assess the potential of rhizobacterial strains to control Verticillium wilt caused by Verticillium dahliae Kleb. METHODS AND RESULTS: Sixty randomly chosen antagonistic bacterial strains originally isolated from rhizosphere of three different host plants of V. dahliae--strawberry, potato and oilseed rape--were evaluated for biocontrol and plant growth promotion by analysing in vitro antagonism towards V. dahliae and other plant pathogenic fungi, production of fungal cell wall-degrading enzymes and plant growth-promoting effects on strawberry seedlings. To test the plant growth-promoting effect, a microplate assay with strawberry seedlings was developed. Although the rhizobacterial strains were isolated from different plants they showed effects on the growth of strawberry seedlings. According to the in vitro biocontrol and plant growth-promoting activity, the three best candidates Pseudomonas putida B E2 (strawberry rhizosphere), Ps. chlororaphis K15 (potato rhizosphere) and Serratia plymuthica R12 (oilseed rape rhizosphere) were selected for greenhouse experiments to verify the in vitro screening results. Under greenhouse conditions the isolates selected according to this strategy were as effective, or more effective than commercial biocontrol agents and may therefore possibly be valuable as antagonists of V. dahliae. CONCLUSIONS: In this study, the screening strategy resulted in a selection of three interesting biocontrol candidates against Verticillium: Ps. putida B E2 (strawberry rhizosphere), Ps. chlororaphis K15 (potato rhizosphere) and Ser. plymuthica R12 (oilseed rape rhizosphere). SIGNIFICANCE AND IMPACT OF THE STUDY: A new combination of in vitro screening methods including a microplate assay with strawberry seedlings to test the plant growth promoting effect which allow to more efficiently select potential biological control agents was developed successfully. PMID- 11851804 TI - The relationships and susceptibilities of some industrial, laboratory and clinical isolates of Pseudomonas aeruginosa to some antibiotics and biocides. AB - AIMS: To provide evidence to support or refute the hypothesis that cross resistance between antibiotics and biocides can occur. METHODS AND RESULTS: Fifty five strains of Pseudomonas aeruginosa were tested for their resistance to anti pseudomonal antibacterials. Twenty clinical, 19 industrial and 16 culture collection isolates were used. The MIC was found for the antibiotics amikacin, ceftazidime, ciprofloxacin, gentamycin, ticarcillin, tobramycin, imipenem and polymyxin B. The MIC was also found for the biocides benzalkonium chloride and chlorhexidine. The analysis of the data was based on the production of a normal distribution of the log (MIC) plots for each antimicrobial. Strains were then labelled as resistant, intermediate or sensitive based on the mean and standard deviation of the distributions. CONCLUSIONS: In general the clinical isolates were the most recalcitrant organisms, with the industrial isolates being the most sensitive. SIGNIFICANCE AND IMPACT OF THE STUDY: The work shows that antibiotic/biocide correlations do occur, especially with clinical strains. That such correlations were not found with industrial isolates suggests that the clinical environment is responsible for the correlation. We could infer that it is the selective pressure of antibiotic usage that differentiates the clinical environment from the industrial. PMID- 11851805 TI - Increased resistance of mice to Salmonella enterica serovar Typhimurium infection by synbiotic administration of Bifidobacteria and transgalactosylated oligosaccharides. AB - AIMS: The anti-infectious activity of Bifidobacteria in combination with transgalactosylated oligosaccharides (TOS) against Salmonella enterica serovar Typhimurium LT-2 in an opportunistic antibiotic-induced murine infection model in mice was examined. METHODS AND RESULTS: B. breve (strain Yakult) with natural resistance to streptomycin sulphate (SM, MIC: > 4 mg ml(-1)), when given daily at a dose of 108 cfu/mouse orally under SM treatment was constantly excreted at 10(10) cfu g(-1) faeces so long as SM was administered, even at 2 weeks after discontinuing administration of B. breve. Explosive intestinal growth and subsequent extra-intestinal translocation of orally infected LT-2 under SM treatment were inhibited by B. breve colonization, and this anti-infectious activity was strengthened by synbiotic administration of TOS with B. breve. Comparison of anti-Salmonella activity among several Bifidobacterium strains with natural resistance to SM revealed that strains such as B. bifidum ATCC 15696 and B. catenulatum ATCC 27539T conferred no activity, even when they reached high population levels similar those of effective strains such as strain Yakult and B. pseudocatenulatum DSM 20439. Both the increase in the concentration of organic acids and the lowered pH in the intestine due to bifidobacterial colonization correlated with the anti-infectious activity. Moreover, the crude cecal extract of B. breve-colonized mice exerted growth-inhibitory activity against LT-2 in vitro, whereas that of the ineffective B. bifidum-colonized cecum showed much lower activity. CONCLUSIONS: Intestinal colonization by bifidobacteria given exogenously together with TOS during antibiotic treatment prevents the antibiotic induced disruption of colonization resistance to oral infection with S. enterica serovar Typhimurium, and the metabolic activity needed to produce organic acids and lower the intestinal pH is important in the anti-infectious activity of synbiotics against enteric infection with Salmonella. SIGNIFICANCE AND IMPACT OF THE STUDY: These results indicate that certain bifidobacteria together with prebiotics may be used for the prophylaxis against opportunistic intestinal infections with antibiotic-resistant pathogens. PMID- 11851806 TI - The gram-positive tonsillar and nasal flora of piglets before and after weaning. AB - AIMS: To investigate gram-positive nasal and tonsillar microbial flora of piglets before and after weaning. METHODS AND RESULTS: The nasal and tonsillar gram positive bacterial flora of 20 non-weaned piglets (2 weeks of age) and 20 weaned piglets (6 weeks of age), obtained from four different piggeries, was quantified by culture and identified by tDNA-PCR. The most widely occurring species from nasal conchae before as well as after weaning in the different piglets investigated were Streptococcus suis and Rothia nasimurium. After weaning a wide variety of Lactobacillus species appeared but in low numbers. In the tonsils, Strep. suis, Strep. dysgalactiae, S. hyicus, S. aureus, Arcanobacterium pyogenes and Actinomyces hyovaginalis were the species isolated from the largest number of pigs before and after weaning. S. aureus and most lactobacilli became more prevalent after weaning. Bacteria not known to be associated with pigs found in the present study included R. nasimurium, Strep. gallolyticus, Pediococcus pentosaceus and some Lactobacillus species. CONCLUSIONS: Over 30 different gram positive bacterial species may occur in nasal conchae and tonsils of unweaned piglets at 2 weeks of age and of 6-week-old weaned piglets. SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrated that weaning is associated with changes in prevalence of only a small minority of the highly diversified bacterial flora of the nares and tonsils of pigs. PMID- 11851807 TI - Detection of Escherichia coli O157:H7 in soil and water using multiplex PCR. AB - AIMS: To evaluate the suitability of a multiplex PCR-based assay for sensitive and rapid detection of Escherichia coli O157:H7 in soil and water. METHODS AND RESULTS: Soil and water samples were spiked with E. coli O157:H7 and subjected to two stages of enrichment prior to multiplex PCR. Detection sensitivities were as high as 1 cfu ml(-1) drinking water and 2 cfu g(-1) soil. Starvation of E. coli O157:H7 for 35 d prior to addition to soil did not affect the ability of the assay to detect initial cell numbers as low as 10 cfu g(-1) soil. Use of an 8-h primary enrichment enabled detection of as few as 6 cfu g(-1) soil, and 10(4) cfu g(-1) soil with a 6-h primary enrichment. When soil was inoculated with 10(5) cfu g(-1), the PCR assay indicated persistence of E. coli O157:H7 during a 35 d incubation. However, when soil was inoculated with lower numbers of pathogen, PCR amplification signals indicated survival to be dependent on cell concentration. CONCLUSIONS: A multiplex PCR-based assay, in combination with an enrichment strategy enabled sensitive and rapid detection of E. coli O157:H7 in soil and water. SIGNIFICANCE AND IMPACT OF THE STUDY: The ability to sensitively detect E.coli O157:H7 in environmental material within one working day represents a considerable advancement over alternative more time-consuming methods for detection of this pathogen. PMID- 11851808 TI - Effect of oregano essential oil on microbiological and physico-chemical attributes of minced meat stored in air and modified atmospheres. AB - AIMS: This study aimed to determine the combined effect of packaging (air, modified atmosphere) with or without the addition of essential oil not only on the selection of microbial association of meat but also to determine any significant difference in microbial metabolites produced from the prevailing bacteria. METHODS AND RESULTS: Samples of minced meat were mixed with different concentration of oregano essential oil (0, 0.05, 0.5 and 1% v/w) and packed under aerobic or with modified atmosphere (Mixed Gas Modified Atmosphere--MGMA, 40% CO2/30% N2/30% O2; or CO2 Modified Atmosphere--COMA, 100% CO2) and stored at 5 degrees C. In all packaging conditions, only concentrations of 0.5% and 1% oregano oil were effective. Inhibition was evident in the order air < MGMA < COMA. Oregano essential oil delayed glucose and lactate consumption aerobically as well as under MGMA. pH changes were also evident. Furthermore, proteolysis was significantly inhibited in aerobically stored samples, and so was the production of acetate under MAP. Similar results were obtained for the other organic acids eluted from HPLC column. CONCLUSIONS: Oregano essential oil delayed microbial growth and suppressed the final counts of the spoilage micro-organisms. It also caused a pronounced alteration in the physico-chemical properties of the minced meat. SIGNIFICANCE AND IMPACT OF THE STUDY: Microbial analysis alone as spoilage index may misrepresent the effect of a hurdle such as essential oils on spoilage. PMID- 11851809 TI - Development of a minimal chemically-defined medium for the exponential growth of Streptococcus thermophilus. AB - AIMS: The present work aimed to define a minimal chemically-defined medium which could sustain the growth of most (if not all) strains of Streptococcus thermophilus. METHODS AND RESULTS: A minimal medium containing 20 components, including one carbohydrate source, six amino acids, two metallic ions, six vitamins and urea allowed for growth of 13 out of 15 Strep. thermophilus strains. Growth of the two last strains required the presence of additional amino acids, the number of which depended on the strain. Growth rates of the strains in the minimal medium ranged from 0.38 to 0.64 h(-1), and final populations were about 10(8) cfu ml(-1). CONCLUSIONS: Streptococcus thermophilus appears much less demanding than other lactic acid bacteria. SIGNIFICANCE AND IMPACT OF THE STUDY: The definition of such a growth medium will be very useful for metabolic flux studies as well as peptide transport studies. PMID- 11851810 TI - Simultaneous detection of Cryptosporidium parvum and Giardia in sewage sludge by IC-PCR. AB - AIMS: The aim of this study was to develop a method based on immunomagnetic capture and polymerase chain reaction (IC-PCR assay) for detection of Cryptosporidium parvum and Giardia intestinalis in sewage sludge. METHODS AND RESULTS: The detection limit of the IC-PCR assay for both organisms was 625 oocysts and cysts ml(-1). By hybridization of PCR products the sensitivity could be increased to 125 oocysts and cysts ml(-1). Forty-four sludge samples from 12 wastewater treatment plants were examined. The samples positive for Giardia (9 out of 44) were from eight wastewater plants and the C. parvum genotype 2 samples (3 out of 44) originated from different sewage works. CONCLUSIONS, SIGNIFICANCE AND IMPACT OF THE STUDY: IC-PCR offers the possibility to distinguish between Cryptosporidium and Giardia genotypes. This assay can be used to monitor the presence of these organisms in a community and to determine contamination of sludge used as soil amendment. PMID- 11851811 TI - The effect of summer and winter seasons on the survival of Salmonella typhimurium and indicator micro-organisms during the storage of solid fraction of pig slurry. AB - AIMS: Investigations were carried out to observe the influence of winter/spring and summer periods on the survival of Salmonella typhimurium and indicator bacteria (psychrophilic, mesophilic, coliform and faecal coliform bacteria and faecal streptococci) in the solid fraction of pig slurry from agricultural wastewater treatment plant. METHODS AND RESULTS: Leather squares and PVC bottles with openings served as test carriers. They were inoculated with broth culture of Salm. typhimurium and introduced directly into the solid fraction. During the experiment, quantitative and qualitative examinations were carried out to determine the presence of Salm. typhimurium and observe the dynamics of indicator bacteria in the solid fraction. CONCLUSIONS: Salmonella typhimurium survived for 26 d in summer and for 85 d in winter/spring. The T90 values of indicator bacteria in summer ranged from 35.44 d (coliform bacteria) up to 100.29 d (mesophilic bacteria). The winter T90 values of indicator bacteria ranged from 74.58 d (faecal coliform bacteria) to 233.07 d (coliform bacteria). SIGNIFICANCE AND IMPACT OF THE STUDY: The present study demonstrated that it is necessary to pay increased attention to the manipulation of slurry solid fraction. PMID- 11851812 TI - A 3.1-kb genomic fragment of Bacillus subtilis encodes the protein inhibiting growth of Xanthomonas oryzae pv. oryzae. AB - AIMS: To clone genes of Bacillus subtilis encoding peptides that inhibit the growth of Xanthomonas orzae pv. oryzae (Xoo). METHODS AND RESULTS: A 3.1-kb DNA fragment from B. subtilis SO113 encoding peptides that inhibit the growth of Xoo (anti-Xoo, showing an inhibition zone) was isolated from a plasmid library of B. subtilis 6 GM15. Sequence analysis revealed that it contained three complete open reading frames (ORFs): ybcO, ybcS and a novel ORF designated ybcPQ. Deleting the last 96 bp of ybcS from the plasmid eliminated the anti-Xoo activity, suggesting that ybcS is required for producing the anti-Xoo activity. However, no anti-Xoo activity could be detected for the plasmid with ybcS alone. Further analysis showed that ybcO, at least, was also required to obtain the anti-Xoo activity. CONCLUSIONS: A fragment of B. subtilis has been cloned that expresses an anti-Xoo activity that requires ybcS and ybcO. SIGNIFICANCE AND IMPACT OF THE STUDY: These genes could be useful for the genetic engineering of resistance to rice bacterial diseases and for the design of new anti-Xoo biocontrol agents. PMID- 11851813 TI - Analysis of the killing of spores of Bacillus subtilis by a new disinfectant, Sterilox. AB - AIMS: To determine the mechanism whereby the new disinfectant Sterilox kills spores of Bacillus subtilis. METHODS AND RESULTS: Bacillus subtilis spores were readily killed by Sterilox and spore resistance to this agent was due in large part to the spore coats. Spore killing by Sterilox was not through DNA damage, released essentially no spore dipicolinic acid and Sterilox-killed spores underwent the early steps in spore germination, including dipicolinic acid release, cortex degradation and initiation of metabolism. However, these germinated spores never swelled and many had altered permeability properties. CONCLUSIONS: We suggest that Sterilox treatment kills dormant spores by oxidatively modifying the inner membrane of the spores such that this membrane becomes non-functional in the germinated spore leading to spore death. SIGNIFICANCE AND IMPACT OF THE STUDY: This work provides information on the mechanism of spore resistance to and spore killing by a new disinfectant. PMID- 11851814 TI - Evaluation of microencapsulation of a Bifidobacterium strain with starch as an approach to prolonging viability during storage. AB - AIMS: To optimize a spray coating process for the production of encapsulated microspheres containing viable Bifidobacterium cells and to determine whether the readily gelatinized modified starch coating used in this study improved bacterial survival in foods or under acid conditions. METHODS AND RESULTS: An air inlet temperature of 100 degrees C was demonstrated to be optimal for the spray drying process, as it afforded good drying, low outlet temperatures (45 degrees C) and resulted in less than 1 log reduction in bifidobacteria numbers during drying. Maximum recovery yields of 30% were obtained after optimizing the air aspiration conditions. The average size of the Bifidobacterium PL1-containing starch microparticles was determined by scanning electron microscopy to be of the order of 5 microm. The starch-coated cells did not display any enhanced viability compared with free PL1 cells when exposed to acid conditions for 6 h or in two dry food preparations over 20 d storage at ambient temperature (19-24 degrees C). Determination of 1491 nucleotides of the 16S rRNA gene from PL1 indicated that it shared 97% homology with a previously sequenced Bifidobacterium ruminantium strain. CONCLUSIONS: Our data demonstrated that, although spray drying is a valuable process for encapsulating bifidobacteria, further work is required to ascertain a more appropriate coating material that will protect this strain against adverse environmental conditions. SIGNIFICANCE AND IMPACT OF THE STUDY: The production of small, uniformly coated microspheres containing viable bifidobacteria using an affordable and industrially convenient process, such as spray drying, has commercial implications for the production of probiotic products. Although popular for use as a coating polymer by the food industry, this study indicated that modified starches might not be suitable for use as an encapsulating material for probiotic strains. PMID- 11851815 TI - Cloning and expression analysis of the 28 kDa protein from Lactobacillus delbrueckii subsp. lactis ATCC 4797 hypothesized to influence lactacin B production. AB - AIMS: A cell wall-associated lactacin B inducer protein (IP) was purified from Lactobacillus delbrueckii subsp. lactis ATCC 4797 (Lact. lactis) by chromatofocusing and gel filtration HPLC (Barefoot et al. 1994). METHODS AND RESULTS: N-terminal sequence of the purified IP was used to design an oligonucleotide (24-mer) for gene identification by Southern and colony hybridizations. Southern hybridization on Lact. lactis chromosomal DNA digested with EcoRI and PstI produced a single 4-5 kbp DNA fragment. Colony hybridizations with 6250 clones produced four positive recombinants for the proposed IP. Sequence of the DNA isolated from RU43e9 revealed a 4623 bp DNA fragment containing three open reading frames (ORF) potentially encoding enzymes that function in glycolysis. One ORF, coding for an active triosephosphate isomerase (Tpi), showed 98% homology to the N-terminal domain of the HPLC purified IP. PCR primers were designed to amplify the ORF encoding the proposed IP for subcloning, protein expression, purification and bacteriocin enhancing assays on pure cultures of Lactobacillus acidophilus N2. CONCLUSIONS: The regions flanking the Tpi gene (data not shown) were also sequenced and it is concluded that the proposed IP reported by Barefoot et al. (1994) is located on an operon containing several glycolytic enzymes that function in glycolysis. SIGNIFICANCE AND IMPACT OF THE STUDY: The findings of this study do not support previously published research (Barefoot et al. 1994) hypothesizing that a purified IP from Lact. lactis, homologous to a Bacillus stearothermophilus Tpi, is capable of enhancing bacteriocin synthesis in Lact. acidophilus N2. PMID- 11851816 TI - Occurrence of Bacillus sporothermodurans and other aerobic spore-forming species in feed concentrate for dairy cattle. AB - AIMS: To determine the aerobic spore composition and presence of Bacillus sporothermodurans spores in feed concentrate for dairy cattle. METHODS AND RESULTS: Six feed concentrate samples from five different farms were analysed. High levels of spores (up to 10(6) spores g(-1)) were found. Identification of 100 selected isolates was obtained by a combination of fatty acid methyl esters analysis, amplified ribosomal DNA restriction analysis and 16S rDNA sequencing. Ninety-seven isolates could be identified to the species level or assigned to a phylogenetic species group. Most of the isolates obtained after a heat treatment of 10 min at 80 degrees C were identified as members of the B. subtilis group (32 isolates), B. pumilus (25 isolates), B. clausii (eight isolates) and B. licheniformis (eight isolates). The isolates with very heat-resistant spores, obtained after a heat treatment of 30 min at 100 degrees C, were identified as members of the B. subtilis group (five isolates), B. sporothermodurans (three isolates), B. amyloliquefaciens (one isolate), B. oleronius (one isolate) and B. pallidus (one isolate). Bacillus cereus was present in each feed concentrate sample and was isolated using a selective mannitol egg yolk polymyxin agar medium. CONCLUSIONS: Feed concentrate for dairy cattle contains known as well as as yet unknown species of Bacillus and related genera with properties relevant to the dairy sector. SIGNIFICANCE AND IMPACT OF THE STUDY: The results formulate the hypothesis that feed concentrate can be a contamination source of spores, including those of B. sporothermodurans, for raw milk at the farm level. PMID- 11851817 TI - Heat and salt stress in the food pathogen Bacillus cereus. AB - AIMS: The effects of stresses imposed on bacterial contaminants during food processing and treatment of packaging material were evaluated on the food pathogen Bacillus cereus. METHODS AND RESULTS: Conditions were established which allowed the cells to adapt to heat, ethanol and hydrogen peroxide stresses, but not to osmotic shock. Cross protection between stresses indicated a clear hierarchy of resistance with salt protecting against hydrogen peroxide, which protected against ethanol, which protected against heat shock. The cultures were shown to be most sensitive to heat, ethanol and oxidative stress at mid exponential phase and to become resistant at stationary phase. Adaptive levels of stressor were found to induce synthesis of general stress and stress-specific proteins and differential accumulation of proteins was demonstrated between heat- or salt-stressed and unstressed cells. CONCLUSIONS: Sequencing revealed that a number of glycolytic enzymes were regulated by heat and osmotic shocks and that the chaperone GroEL was induced by heat shock. SIGNIFICANCE AND IMPACT OF THE STUDY: The implications of the physiological data in designing storage and processing conditions for food are discussed. The identification of stress regulated proteins reveals a clear role for glycolysis in adaptation to heat shock and osmotic stress. PMID- 11851818 TI - Resuscitation rate in different enterococcal species in the viable but non culturable state. AB - AIMS: The viable but non-culturable (VBNC) state is a survival strategy adopted by bacteria when exposed to environmental stress. When in this state bacteria are no longer culturable on conventional growth media, but cells display metabolic activity and maintain pathogenicity factors/genes and, in some cases, resuscitation from the VBNC state has been shown. This state has been described for both human pathogens and faecal pollution indicators. In this study, we present evidence for entry of different enterococcal species into the VBNC state in an oligotrophic microcosm. METHODS AND RESULTS: The duration of the viability of the cells in the VBNC state was measured either by detecting the presence of pbp5 mRNA or by quantifying their resuscitation capability. Enterococci showed different behaviours. Enterococcus faecalis and Enterococcus hirae entered into the VBNC state within 2 weeks and remained in that state for 3 months. In the experiments described the resuscitation rate was 1:10 000 cells as soon as the cells entered the VBNC state and decreased gradually to undetectable levels over the following 3 months. Enterococcus faecium, however, remained culturable up to 4 weeks. After this time period, when the population was totally unculturable, the cells were far less resuscitable than other enterococci and only over a narrow time interval (2 weeks). CONCLUSIONS: These results suggest that Ent. faecalis and Ent. hirae enter the VBNC state but that Ent. faecium, in an oligotrophic laboratory environment, tends to die instead of entering the VBNC state. SIGNIFICANCE AND IMPACT OF THE STUDY: These experiments may mimic what happens when enterococci are released by humans and animals in natural environments. PMID- 11851819 TI - Usefulness of epifluorescence for quantitative analysis of lactobacilli in probiotic feed. AB - AIMS: Enumeration of total, active or viable probiotic micro-organisms from liquid or solid commercial feedstuffs was studied during processing and storage. METHODS AND RESULTS: After sample preparation, an epifluorescence microscopy technique and a plating method were investigated comparatively. It was shown that (i) on the day of manufacture, active or viable bacteria were in equivalent amounts and that viable numbers then decreased, depending on the different processing and storage factors enhancing ABNC production, (ii) the amount of total and active lactobacilli remained close and quite stable for months at a high level (>10(8) active fluorescent units). CONCLUSIONS: Processing and storage promoted ABNC cells in the products tested. Consequently, both techniques should be used to evaluate the viable-dead-active status of bacteria for which functional properties are claimed. SIGNIFICANCE AND IMPACT OF THE STUDY: Enumeration of the whole probiotic bacterial population should be take into account for guidelines and labelling since non-viable bacteria could have a probiotic effect. PMID- 11851820 TI - The regulatory effects of whey retentate from bifidobacteria fermented milk on the microbiota of the Simulator of the Human Intestinal Microbial Ecosystem (SHIME). AB - AIMS: To investigate the effects of whey retentate from Bifidobacteria fermented milk. METHODS AND RESULTS: The simulator of the human intestinal microbial ecosystem (SHIME) was used. The composition of the microbiota and its metabolic activities were analysed. Changes in the microbial composition became apparent within 15 days of the treatment in the vessels representing the ileum and the large intestine. The whey retentate favoured the growth of endogenous bifidobacteria and induced a decrease in Bacteroides fragilis and in sulphite reducing clostridia, especially Clostridium perfringens. After the administration was stopped, these populations tended to revert to their original levels, except for the streptococci and the staphylococci populations. The treatment also led to an increase in acetic acid, CH4 and CO2 production, suggesting overgrowth of some anaerobic bacteria. Ammonium, generally considered as undesirable, declined. CONCLUSIONS: The whey retentate clearly altered the microbial community in the SHIME. SIGNIFICANCE AND IMPACT OF THE STUDY: Whey retentate appears to exert a beneficial effect on the in vitro gastrointestinal system; these findings warrant confirmation by in vivo studies. PMID- 11851821 TI - Synthesis and evaluation of novel fluorogenic substrates for the detection of bacterial beta-galactosidase. AB - AIMS: A widely used coumarin derivative is 7-hydroxy-4-methylcoumarin-beta-D galactoside (4-methylumbelliferone-beta-D-galactoside; 4-MU-GAL). This galactoside is utilized as a substrate for the detection of the beta galactosidase activity of coliform bacteria in water analysis. The intense fluorescence of coumarin-based molecules has enabled them to be incorporated into enzyme-based tests for the quantitative assay of indicator bacteria. The aim of this present study was to evaluate the potential of other coumarin derivatives, by synthesis of a selection of core coumarin molecules. METHODS AND RESULTS: Several coumarin derivatives were found to be more promising than 4-MU, with ethyl-7-hydroxycoumarin-3-carboxylate (EHC) giving a combination of greater fluorescence over a broad pH range and reduced growth inhibition with 12 representative coliform strains. On conversion to a beta-galactoside derivative, EHC-GAL generated a more rapid fluorescence than any other tested substrate. CONCLUSIONS: When tested in a broth assay format, based on most probable number (MPN), low numbers of coliforms were detected with EHC-GAL around 1 h earlier than with 4-MU-GAL. SIGNIFICANCE AND IMPACT OF THE STUDY: The present study suggests that EHC-GAL should be evaluated as a substrate for the detection of coliforms in water analysis, due to a combination of the following favourable features: (i) reduced toxicity; (ii) increased fluorescence; (iii) pH stability of fluorescence; and (iv) rapid detection. PMID- 11851822 TI - Growth optimization of Pediococcus damnosus NCFB 1832 and the influence of pH and nutrients on the production of pediocin PD-1. AB - AIMS: Optimization of the growth of Pediococcus damnosus NCFB 1832 and the production of pediocin PD-1 by traditional fermentation methods. METHODS AND RESULTS: Fermentation studies were conducted in De Man Rogosa and Sharpe (MRS) broth (Oxoid), preadjusted to specific pH values, and in MRS broth supplemented with various nitrogen sources, MnSO4, MgSO4 and Tween 80. The production of pediocin PD-1 closely followed the growth curve of Ped. damnosus NCFB 1832. Maximum levels of bacteriocin activity (3249 AU ml(-1)/O.D.max) were recorded in MRS broth with an initial pH of 6.7. In media with an initial pH of 4.5 bacteriocin activity as low as 222 AU ml(-1)/O.D.max was recorded. The highest bacteriocin activity was recorded in growth conditions allowing the greatest pH variation (highest DeltapH). The addition of bacteriological peptone (1.7%, w/v), MnSO4 (0.014%, w/v) and Tween 80 (3%, v/v) to MRS and adjustment of the medium pH to 6.7 resulted in a further increase in activity (from 3249 to 5078 AU ml( 1)/O.D.max). The same medium, but with an initial pH of 6.2, resulted in an 82.5% decrease in bacteriocin activity. CONCLUSIONS: Pediocin PD-1 production is not only stimulated by the presence of specific growth factors (e.g., bacteriological peptone, MnSO4 or Tween 80), but may also be stimulated by the lowering in pH during growth (highest DeltapH), and thus also the amount of organic acids produced. SIGNIFICANCE AND IMPACT OF THE STUDY: The production of pediocin PD-1 by the wild-type producer strain was significantly improved by using a defined medium and traditional fermentation methods. PMID- 11851825 TI - Hepatic hydroxylation of bile acids and chronic liver diseases: do transporters play a mechanistic role? PMID- 11851826 TI - Primary biliary cirrhosis and pernicious anemia. PMID- 11851827 TI - Benign nodular hepatocellular lesions caused by abnormal hepatic circulation: etiological analysis and introduction of a new concept. AB - Problems in definitive diagnosis and etiology of various benign nodular hepatocellular lesions were evaluated. Of these lesions, focal nodular hyperplasia (FNH), nodular regenerative hyperplasia (NRH), nodular lesions associated with idiopathic portal hypertension (IPH), non-cirrhotic large regenerative nodules (LRN), hepatocellular adenoma (HA)-like hyperplastic nodules, and partial nodular transformation (PNT) have been suggested to be related to abnormal hepatic circulation. However, the following points are considered to need further clarification: (i) is the abnormal circulation caused by thrombosis, vasculitis, or congenital anomaly?; (ii) is thrombosis a cause or a result of congestion?; (iii) are impaired blood vessels primarily the portal veins or arteries?; (iv) how are these disorders related to various syndromes, immunological abnormalities and abnormal blood flow of other organs, which are reported to coexist with these lesions often?; and (v) how should non-typical cases, which differ from typical cases, be interpreted? In addition, a concept that may lead to solving these problems (anomalous portal tract syndrome; a hypothesis that congenital vascular anomaly is the origin of these benign nodular hepatocellular lesions) was introduced. PMID- 11851828 TI - Chemoprevention of hepatocellular carcinoma: concept, progress and perspectives. AB - Hepatocellular carcinoma (HCC) often develops in patients with chronic liver diseases associated with hepatitis B (HBV) and hepatitis C (HCV) virus infections with high incidences. Particularly, post-therapeutic recurrence encountered after the curative treatment of the preceding HCC may limit the prognosis. Thus, prevention of HCC is of great significance. In the present review, immunopreventions with alpha-interferon and glycyrrhizin, as well as chemoprevention with acyclic retinoid, are discussed. alpha-Interferon prevents the development of HCC not only in patients with a long-term elimination of HCV (sustained virological responders), but in ones with normalized serum aminotransferases (sustained biochemical responders). Glycyrrhizin also suppresses serum aminotransferases and thereby prevents the tumor development, even though the compound does not have antiviral activity for HBV or HCV by itself. Therefore, suppression of hepatic necroinflammation by these drugs may serve to prevent hepatocarcinogenesis. In contrast, acyclic retinoid suppresses the post-therapeutic recurrence in cirrhotic patients who underwent curative treatment of preceding tumors. The retinoid induces the disappearance of serum lectin-reactive alpha-fetoprotein (AFP-L3), a tumor marker indicating the presence of unrecognizable tumors in the remnant liver, suggesting a deletion of such minute (pre)malignant clones (clonal deletion). As a molecular mechanism of the clonal deletion, a novel mechanism of apoptosis induction by the retinoid via tissue transglutaminase is implicated. In future, a combination of immunopreventive and chemopreventive therapies may give a clue to the further advances of cancer prevention, and thereby to the improvement of the prognosis of cirrhotic patients. PMID- 11851829 TI - Reactive oxygen species activity, mucosal lipoperoxidation and glutathione in Helicobacter pylori-infected gastric mucosa. AB - BACKGROUND AND AIM: Helicobacter pylori is considered as the major pathogen in Helicobacter pylori-associated gastroduodenal disease, but the mechanism of its action has not been fully explained. This study was performed to assess the reactive oxygen species activity and the damage in Helicobacter pylori-infected gastric mucosa. METHODS: Gastric biopsy specimens were obtained from 308 patients undergoing endoscopy. Gastric mucosal damage was assessed by using luminol enhanced chemiluminescence, thiobarbituric acid-reactive substance, and mucosal glutathione. RESULTS: The chemiluminescence and thiobarbituric acid-reactive substance-equivalent levels in the mucosa of patients with Helicobacter pylori positive gastric mucosa (43.8 +/- 134.9 c.p.m./microg tissue, 157.0 +/- 96.2 nmol/g tissue, respectively) were significantly higher than in those with Helicobacter pylori-negative mucosa (6.8 +/- 20.3 c.p.m./microg tissue, 110.0 +/- 51.6 nmol/g tissue, respectively; P=0.000, P=0.016, respectively). The glutathione levels in the mucosa of patients with Helicobacter pylori-positive gastric mucosa (159.3 +/- 76.6 nmol/microg tissue) were significantly lower than in those with Helicobacter pylori-negative gastric mucosa (212.3 +/- 134.3 nmol/microg tissue; P=0.008). After the data were divided according to the presence of Helicobacter pylori, there were no significant differences in chemiluminescence, thiobarbituric acid-reactive substance, and glutathione among the different macroscopic findings within Helicobacter pylori-positive and negative gastric mucosa. CONCLUSIONS: Helicobacter pylori infection plays a pathological role in many gastrointestinal diseases through excessive mucosal reactive oxygen species production, pronounced membrane damage, and the depletion of gastric anti-oxidants. PMID- 11851830 TI - Extending the reading time increases the accuracy of rapid whole blood test for diagnosis of Helicobacter pylori infection. AB - BACKGROUND: To evaluate the accuracy of two rapid whole blood tests (the BM-Test Helicobacter pylori and the QuikPac IV One Step H. pylori Whole Blood Test), and compare this to a conventional quantitative ELISA test (HEL-p TEST II). METHODS: Helicobacter pylori status in dyspeptic patients was assessed by culture, histology, and rapid urease tests on biopsies from the antrum and corpus. The optimal cut-off value of the reading time for the rapid blood tests was determined by using the receiver characteristics operative (ROC) curves. RESULTS: In the 141 patients examined, 89 were infected, 51 were not infected, and one was indeterminate (only positive in either urease test or histology). Areas under ROC curves were greater in the BM-Test compared with the QuikPac IV (0.948 vs 0.840, P < 0.01), with their most appropriate cut-off reading times at 360 and 395 min, respectively, rather than 10 min as suggested by the manufacturer. The sensitivity and specificity were 94.4% and 94.1% at 360 min, and 74.2 and 96.1% at 10 min for the BM-Test; 80.9, 76.5 at 395 min and 3.4 and 100% at 10 min for the QuikPac IV. The antibody titer of the quantitative ELISA test was negatively correlated with the reaction time of the two rapid blood tests in H. pylori infected patients (P < 0.05, r=-0.3). CONCLUSIONS: The BM-Test is an appropriate office-based test for diagnosing H. pylori infection in Chinese patients. Extending the reading time would facilitate the readability of rapid blood tests with a resultant increase in accuracy. PMID- 11851831 TI - Endoscopically demonstrable esophageal changes after Helicobacter pylori eradication in patients with gastric disease. AB - BACKGROUND AND AIMS: An increased prevalence of reflux esophagitis has been reported following Helicobacter pylori (H. pylori) eradication in patients with duodenal ulcers in Western countries. However, it has remained unknown whether this might also appertain to individuals with other diseases. We therefore carried out this study to determine the effect of eradicating H. pylori infection in a series of Japanese patients. METHODS: Of a total of 203 H. pylori-positive patients successfully cured of infection, 82 cases (58 males, 24 females) with gastric disease, but not duodenal ulcers, were included in the present study; median age 56 years (range 18-80) and median follow up of 24 months (range 6-65). The patients were investigated clinically and endoscopically at regular intervals. RESULTS: Mild reflux esophagitis developed after eradication in three of 55 (5.5%) patients formerly without this condition, while it improved after eradication in five of 27 (18.5%) patients, with the disease endoscopically diagnosed prior to eradication. The estimated incidence of esophagitis within 3 years was 4.8% after cure of infection. Short segment Barrett's esophagus developed after eradication in six of 58 (10.3%) patients who did not have it prior to the therapy, while the condition did not improve in 24 patients affected before eradication. CONCLUSIONS: Endoscopic esophageal changes after H. pylori eradication in the present series of Japanese patients were relatively infrequent and mild. This therapeutic approach thus appears to be safe and unproblematic. PMID- 11851832 TI - Spatial distribution and histogenesis of colorectal Paneth cell metaplasia in idiopathic inflammatory bowel disease. AB - BACKGROUND AND AIM: Colorectal Paneth cell metaplasia (PCM) is known to be a sign of idiopathic inflammatory bowel disease (IBD), although its distribution and histogenesis are not fully understood. Objectives of this research were to investigate the spatial distribution of PCM in IBD and other forms of colitis (non-IBD), and to find stimuli causing PCM. METHODS: We studied multiple biopsy specimens from 181 patients with ulcerative colitis (UC), 159 with Crohn's disease (CD), 448 with non-IBD, and 78 normal controls. Paneth cell metaplasia frequency, at each colorectal site, was evaluated to find possible differences among diseases, phases of activity, and extents of disease. RESULTS: In non-IBD and controls, PCM was rarely (0-1.9%) seen at distal sites, but frequently (up to 48.7%) found at the ascending colon and cecum (P < 0.001). Paneth cell metaplasia frequency was significantly higher in IBD than in non-IBD patients and controls at distal sites (P < 0.001), but did not differ significantly between UC and CD, or among active, resolving, and quiescent phases. In UC, proctitis and left-sided colitis rarely displayed PCM at unaffected sites. Multiple logistic regression analysis revealed that PCM was positively associated with crypt distortion and mononuclear cell infiltration (P < 0.005), but negatively or not significantly associated with crypt atrophy, mucin depletion, acute inflammation, or phase of activity. CONCLUSIONS: Paneth cell metaplasia is a non-specific phenomenon in the proximal colon, but distal PCM, which occurs exclusively in affected mucosa, is a useful marker indicating IBD, even in the inactive phase. Regression analysis suggests that repair and regeneration may be the most potent stimuli causing PCM. PMID- 11851833 TI - Correlation of rectum-associated lymph nodules with the development of experimentally induced acute colonic inflammation in rats. AB - BACKGROUND AND AIMS: The mechanisms of inflammation in ulcerative colitis occurring initially in the rectum and then extending throughout the colon are still unknown. The aim of this study is to investigate the correlation of rectum associated lymph nodules (RALN) and the development of a dextran sulfate sodium induced experimentally acute colonic inflammation in rats. METHODS: We immunohistochemically analyzed the lymph nodules in the rectal region of rats using monoclonal antibodies to specific markers. RESULTS: Rectum-associated lymph nodules resembled the Peyer's patches, where CD4+, CD8+ lymphocytes and Mar 1+ macrophages were observed. After the administration of dextran-sulfate sodium (DSS), RALN showed hypertrophy with an increase in the number of CD4+, CD8+, and interleukin (IL)-2R+ lymphocytes in the periphery, as well as Mar 1+ macrophages in the entire region. Concurrently, platelet/endothelin cell adhesion molecule-1 (PECAM-1+) blood vessels, including high-endothelial venules (HEV), were increased in number in the interfollicular areas. Furthermore, a number of small lymph nodules varying in size were observed in the upper region of the rectum. Some of them were initially confined to the lamina propria mucosae, and became large enough to extend deeper into the tela submucosa. These DSS-induced lymph nodules contained large numbers of CD4+ lymphocytes, IL-2R+ lymphocytes, and Mar 1+ macrophages. CONCLUSIONS: The pathological changes of ulcerative colitis were well correlated with the development of both RALN and DSS-induced lymph nodules. The immunological reaction that occurred in DSS-induced lymph nodules is significantly associated with the expansion of colitis from the lower region of rectum to the upper region of rectum and colon. PMID- 11851834 TI - Observer variation of diagnoses based on simple biopsy criteria differentiating among Crohn's disease, ulcerative colitis, and other forms of colitis. AB - BACKGROUND AND AIM: Simple mucosal biopsy criteria proposed by authors reliably differentiate idiopathic inflammatory bowel disease (IBD) from other forms of colitis (non-IBD) and Crohn's disease involving the colon (CD) from ulcerative colitis (UC). The aim of this study is to investigate the reproducibility of these criteria. METHODS: Three established pathologists and two medical students blindly examined 20 sets of multiple biopsy slides from patients with CD, 20 from those with UC, and 20 from those with non-IBD. The students had been given instructions on histological definitions using another 15 sets previously. Each observer evaluated 10 histological items required in the criteria and determined categorical diagnoses such as definite IBD and probable UC. Interobserver agreement for the individual histological items was measured by using kappa analysis and Pearson's correlation, while it was measured for categorical diagnoses with the use of Spearman's rank correlation. RESULTS: All of the individual histological items expressed excellent or fair-to-good agreement among the five observers, although two items associated with the criteria for CD and UC had poor agreements among the students. With regard to categorical diagnoses based on the criteria for IBD and non-IBD, and those for CD and UC, coefficients for Spearman's rank correlation exceeded 0.92 and 0.86 among the pathologists, and 0.76 and 0.74 among the students, respectively; all of the coefficients were statistically significant (P < 0.05). CONCLUSIONS: The simple criteria were sufficiently reproducible and would help most pathologists to make an automated and objective diagnosis. PMID- 11851835 TI - Prevalence of vaccine-induced escape mutants of hepatitis B virus in the adult population in China: a prospective study in 176 restaurant employees. AB - BACKGROUND AND AIM: Hepatitis B virus (HBV) variants with mutations in the S gene would pose a substantial risk to the community as current HBV vaccines are not effective in preventing infection with them. The majority of such vaccine escape mutants so far reported have been found while studying vertical transmission of HBV; the vaccine failure rate in connection with vaccine escape mutants in adults is not clear at the moment. The purpose of this study was to evaluate the efficacy of immunization against HBV in the adult population by analysis using polymerase chain reaction (PCR) to detect HBV-DNA, and also to elucidate the type of mutation encountered in vaccine failure cases. METHOD: A total of 176 adult restaurant employees in China, who had been vaccinated according to the food epidemic law, were enrolled in a standard vaccination program. Their serum HBV DNA was determined before and 1 year after the completion of the vaccination program. In those infected with HBV, despite having received the HBV vaccine, direct sequencing within the S gene of the amplified samples was conducted. RESULTS: Although only two cases were found to be hepatitis B surface antigen (HBsAg) positive 1 year after the completion of the vaccination program, six subjects (3.4%) were found to be HBV-DNA positive assessed by a nested PCR. Four out of these six cases had a point mutation within the 'a' determinant; they were Gly-145-Ala, and Ile/Thr-126-Asn/Ser. CONCLUSION: The HBV vaccine failure rate assessed by using PCR analysis was 3.4% (six of 176) in the Chinese adult population undergoing the HBV vaccination program. Hepatitis B virus variants with missense mutation within the 'a' determinant were responsible in most cases. PMID- 11851836 TI - Multicenter prospective analysis of newly diagnosed hepatocellular carcinoma with respect to the percentage of Lens culinaris agglutinin-reactive alpha fetoprotein. AB - BACKGROUND AND AIM: The Lens culinaris agglutinin-reactive fraction of alpha fetoprotein (AFP-L3) has been reported to be a highly useful marker for hepatocellular carcinoma (HCC) compared with a conventional serum AFP concentration, which allows earlier detection of HCC compared with using other imaging modalities and predicting prognosis after therapy. A collaborative prospective study involving nine Japanese hospitals was conducted to analyze the relationships between the tumor characteristics of a HCC patient and the percentage of AFP-L3/AFP total at the initial detection. METHODS: Between 1 October 1996 and 30 September 1997, a total of 388 patients with newly diagnosed HCC were registered. RESULTS: The cut-off level of the percentage of AFP-L3 was altered from 15 to 10%. The AFP-L3-positive HCC patients demonstrated the characteristics of having an advanced tumor, such as the number of tumors, maximum diameter, tumor spread, portal vein invasion, tumor stage, and tumor classification. With the conventional cut-off level of 15% of the percentage of AFP-L3, the malignant characteristics were more definite than that of 10%. However, no significant differences of serum AFP concentration were observed for malignant characteristics such as maximum diameter and histopathological grading. CONCLUSION: Serum AFP concentration does not reveal a malignancy of HCC, however, the AFP-L3-positive HCC has biologically malignant characteristics, especially portal vein invasion and lower tumor classification, and is an advanced tumor regardless of small tumor size and lower serum AFP concentration. As AFP-L3 shows the tumor characteristics, its presence should be an important factor in the determination of therapy and prognosis of patients. PMID- 11851837 TI - Autoimmune hepatitis in Singapore: a rare syndrome affecting middle-aged women. AB - BACKGROUND AND AIM: The prevalence of autoimmune hepatitis in Singapore is unknown. Over a period of 7 years, 24 cases were diagnosed in a district general hospital in Singapore (Toa Payoh Hospital) by using the scoring system proposed by the International Autoimmune Hepatitis Group in 1993. The aims of our study were to determine the prevalence of autoimmune hepatitis in Singapore, and to investigate the characteristics and prognosis in the mainly Chinese population. METHODS: The case records of all 24 patients were reviewed, and the following parameters were recorded: age at presentation, sex, symptoms and signs at presentation, past exposure to hepatotoxic drugs, alcohol intake, blood transfusion laboratory and histological tests used to determine autoimmune hepatitis, response to treatment, complications, and survival. RESULTS: The mean age of patients was 57 years old. There was a female-male ratio of 11:1. Forty two percent of the patients were cirrhotic at presentation. The prevalence of autoimmune hepatitis was four per 100 000, with no significant difference between Chinese, Malay and Indian patients (Odds ratio of 0.38 by the chi-squared test). Eighty-nine percent of the patients responded to treatment with the induction of prednisolone, but the relapse rate was 61%. Treatment failure occurred in one patient. The mortality rate during the 7 years of follow up was 21%, and all were caused by complications of cirrhosis. The survival at 5 years was 71%, with a standard error of 0.13. CONCLUSION: Autoimmune hepatitis in Singapore is mainly a disease in older women. The response to steroid treatment is good, with a 5-year survival rate of 71%. PMID- 11851838 TI - Antimitochondrial autoantibodies in saliva and sera from patients with primary biliary cirrhosis. AB - BACKGROUND AND AIMS: Primary biliary cirrhosis (PBC) is a cholestatic autoimmune liver disease characterized by antimitochondrial autoantibodies (AMA) in serum, for which the reactants are E2 subunits of the three 2-oxoacid dehydrogenase (2 OAD) enzymes, particularly pyruvate dehydrogenase complex (PDC-E2). Some 70% of patients with PBC have a coexisting autoimmune disease including Sjogren's syndrome. We aimed to ascertain the frequency and isotype of AMA in saliva in PBC. METHODS: Serum and saliva from 12 patients with PBC were tested for AMA by immunoblotting on bovine heart mitochondria, and by an automated microassay based on inhibition of the enzymatic activity of PDC. RESULTS: Autoantibodies of the immunoglobulin (Ig)G, IgM, and IgA immunoglobulin isotypes against the E2 subunits of 2-OAD enzymes were demonstrable in PBC in serum (12 of 12 cases) and saliva (nine of 12 cases). Salivary autoantibodies, like serum autoantibodies, were predominantly reactive with PDC and of the IgG isotype. Results for serum and saliva corresponded closely with regard to reactivity with individual enzymes of the 2-OAD enzyme family, and to the autoantibody isotype that was predominantly expressed, and also in the capacity to inhibit the enzymatic activity of PDC. CONCLUSIONS: The presence of AMA in saliva to 2-OAD enzymes indicates that salivary glands could participate in the pathogenetic process of PBC. The detection of salivary AMA by a semi-automated enzyme inhibition assay offers possibilities for rapid population screening for detection of preclinical PBC among at-risk individuals, middle-aged to older women. PMID- 11851839 TI - Role of non-alcoholic steatohepatitis in methotrexate-induced liver injury. AB - BACKGROUND AND AIMS: Hepatotoxicity, especially liver fibrosis, is the major concern with long-term, 'low-dose' oral methotrexate (MTX) therapy for psoriasis. The histological features are non-specific and resemble those of non-alcoholic steatohepatitis (NASH). Moreover, most of the risk factors of MTX-induced liver injury are also associated with NASH. In this study, we investigate whether NASH contributes to the prevalence and progression of MTX-induced liver injury in patients receiving MTX for psoriasis. METHODS: Clinical details, including MTX dosage schedules and risk factors for liver injury, was documented for 24 patients on long-term MTX therapy for psoriasis. Serial liver biopsies were graded according to the Roenigk classification scale and a recently proposed grading and staging system for NASH. RESULTS: Thirteen of the 17 patients who had a NASH-like pattern of liver injury also had the risk factors for NASH obesity and/or diabetes, and all had progressive liver injury. The other four patients had no risk factors, but a mean cumulative dose of 6.5 g. Seven patients, who did not have a NASH-like pattern of injury, had a mean cumulative dose of 3.8 g. There was a positive correlation between the cumulative dose, risk factors and progression when the biopsies were scored by the modified grading and staging classification for NASH, but not with the Roenigk system. CONCLUSIONS: Non steatohepatitis, probably aggravated by MTX, is an important cause of liver injury in patients on long-term, 'low-dose' MTX treatment for psoriasis. In addition, MTX alone can cause a NASH-like pattern of injury that is at least, in part, caused by a higher cumulative dose. PMID- 11851840 TI - Glucose-6-phosphatase gene mutations in 20 adult Japanese patients with glycogen storage disease type 1a with reference to hepatic tumors. AB - BACKGROUND AND AIMS: A few cases are reported of liver neoplasms observed in patients with glycogen storage disease type 1a (GSD1a). Genetic analysis was carried out in adult Japanese patients with GSD1a and their family members, and hepatic tumors were also investigated in these patients. METHODS: DNA was extracted from the peripheral blood lymphocytes of 20 adult patients with GSD1a and 21 family members, and mutations were detected based on the differences in the polymerase chain reaction (PCR) products of the glucose-6-phosphatase (G6Pase) gene shown by single-strand conformation polymorphism (SSCP) analysis. Actual mutations were confirmed by direct sequencing. The relationship between the occurrence of liver tumors and the clinical characteristics of the patients was also investigated. RESULTS: Nineteen of the 20 patients were homozygous for the G727T mutation and one was a compound heterozygote for G727T plus G327A mutations. All of the 19 homozygotes for G727T had hepatomegaly, three had hepatocellular carcinoma, one had cholangiocellular carcinoma, and seven had hepatic adenoma. There were no differences between the tumor and non-tumor groups with respect to laboratory biochemical data (P > 0.05). The mean age of G727T homozygotes with hepatocellular carcinoma was 48.3 years, and that of those with hepatic adenoma was approximately 20 years younger. CONCLUSION: The G727T mutation seems to be common among Japanese patients with GSD1a, and the discovery of one heterozygote with a combination of G727T and G327A mutations (the latter mutation is common among Chinese) by the use of polymerase chain reaction-single strand conformation polymorphism analysis gave further insight into Japanese ancestry. This is the first study of liver tumors in a large group of adult GSD1a patients with the G727T mutation. As most of the patients in our series are free from other chronic liver diseases such as viral hepatitis, other genetic and/or acquired factors may have influence on the sequel to this metabolic disease. PMID- 11851841 TI - Inverse relationship between circulating levels of leptin and bone mineral density in chronic liver disease. AB - BACKGROUND AND AIM: The pathophysiology of osteoporosis complicating chronic liver disease is unknown. Recent animal studies have found leptin to be a potent inhibitor of bone formation. The aim of this study was to investigate the relationship between serum leptin levels and bone mineral density in patients with chronic liver disease. METHODS: Fifty-eight patients, 39 females and 19 males, and age- and gender-matched controls were included. Bone mineral density was measured by using dual energy X-ray absorptiometry. Serum leptin was measured by using a radioimmunoassay. RESULTS: The mean serum leptin concentration was 10.4 +/- 11.3 and 15.2 +/- 17.9 ng/mL; P=0.11, in the patients and controls, respectively. Leptin correlated positively with body mass index in patients (r=0.40; P=0.003) and in controls (r=0.55; P < 0.0001). In patients classified as Child-Pugh grade B and C, serum leptin correlated negatively with bone mineral density in females at both the lumbar spine and the femoral neck (r=-0.78; P=0.04 and r=-0.86; P=0.03, respectively). In male patients, the correlation was only significant at the lumbar spine (r=-0.99; P=0.002 and r=-0.86; P=0.06, at the lumbar spine and femoral neck, respectively). No correlation was found between serum leptin and bone mineral density in the controls. CONCLUSION: An inverse relationship between serum leptin and bone mineral density was found in patients with advanced chronic liver disease. The reasons for these findings are uncertain, but a pathophysiological role of circulating leptin in osteoporosis in chronic liver disease is possible. PMID- 11851842 TI - Hepatobiliary and pancreatic: a woman with jaundice and anorexia. PMID- 11851843 TI - Gastrointestinal: celiac disease. PMID- 11851845 TI - A female with asymptomatic primary biliary cirrhosis associated with pernicious anemia. AB - We experienced a female case with asymptomatic primary biliary cirrhosis that was associated with pernicious anemia after 16 years from the onset. She was 52 years old when she first visited a clinic in 1981 for liver dysfunction treatment. Antimitochondrial antibody was negative and antipyruvate dehydrogenase complex antibody was positive in a low titer in its immunoglobulin (Ig)M type. Histological examination of her liver revealed a presence of definite chronic non suppurative destructive cholangitis with numerous epithelioid cell granuloma. She had been given 600 mg of the oral daily dose of ursodeoxycholic acid since 1992. Macrocytic anemia incidiously appeared in September 1999. An immunological examination detected negative antiparietal cell antibodies and positive anti intrinsic factor antibodies. Her bone marrow smear showed numerous megaloblasts and serum vitamin B12 in her blood was low at 99 pg/mL. Severe reversed atrophic type gastritis (type A gastritis) was demonstrated by the use of dye-endoscopy with Congo red. Her macrocytic anemia dramatically improved after intramuscular administration of vitamin B12. In conclusion, attention should be given to the association of pernicious anemia during the follow up of primary biliary cirrhosis. PMID- 11851846 TI - Embolization of the left portal vein to inferior vena cava shunts for chronic recurrent hepatic encephalopathy via the mesenteric vein. AB - Diagnostic imaging and embolization therapy for very rare intrahepatic portal systemic shunts with liver cirrhosis are reported. An 82-year-old woman was admitted to the hospital (Yachiyo Hospital) because of hepatic encephalopathy. Computed tomography with contrast enhancement demonstrated anomalous vessels between the portal vein and the inferior vena cava. Those shunts were suspected as the cause of her encephalopathy with hyperammonemia. Portography through McBurney's laparotomy demonstrated two portal-caval shunts; one was from the bifurcation of the portal vein and the other was from the left portal vein. They seemed to originate from the vascular system of the caudate lobe, and were obstructed with stainless coils. The patient is well with a normal serum ammonia level 40 months following the intervention. PMID- 11851847 TI - Extensive mesenteric vein and portal vein thrombosis successfully treated by thrombolysis and anticoagulation. AB - Mesenteric vein thrombosis is generally difficult to diagnose and can be fatal. A case of extensive thrombosis of the mesenteric and portal veins was diagnosed early and successfully treated in a 26-year-old man with Down syndrome who was admitted to hospital because of abdominal pain, severe nausea and high fever. Ultrasonography revealed moderate ascites, and there was minimal flow in the portal vein (PV) on the Doppler examination. Computed tomography (CT) showed remarkable thickening of the walls of the small intestine and extensive thrombosis of the mesenteric, portal and splenic veins. Because neither intestinal infarction nor peritonitis was seen, combined thrombolysis and anticoagulation therapy without surgical treatment was chosen. Urokinase was administered intravenously and later through a catheter in the superior mesenteric artery. Heparin and antibiotics were given concomitantly. The patient's symptoms and clinical data improved gradually. After 10 days, CT revealed that collateral veins had developed and the thrombi in the distal portions of the mesenteric veins had dissolved, although the main trunk of the PV had not recanalized. The only risk factor of thrombosis that was detected was decreased protein S activity. PMID- 11851851 TI - The good life for citizens with intellectual disability. PMID- 11851852 TI - Injury risk in young people with intellectual disability. AB - Epidemiological studies have established injury as a major public health problem for young people in the general population, but minimal population-based injury information exists on those with intellectual disability (ID). The present study investigated risk factors for injury to subjects in the Australian Child and Adolescent Development (ACAD) programme, which is examining emotional and behavioural problems in young people with ID. Extensive biopsychosocial data for the ACAD programme were collected at two time intervals (i.e. 1990-1991 and 1995 1996). Carer report of medically attended injury over one year was collected for the first time period (1995-1996) on a sample of 465 ACAD subjects (aged 5-29 years) representative of young people with ID (IQ < 70) to determine risk factors for injury using the ACAD biopsychosocial data. Psychopathology [odds ratio (OR)=3.4] epilepsy (OR = 2.4) and an overly sociable temperament (OR=2.2) are associated with injury in young people with ID. Social and family factors had minimal influence on injury risk in this population. Those who are highly disruptive, self-absorbed, anxious, have problems relating socially, have communication disturbance or have attention deficit hyperactivity symptoms are particularly at increased risk for injury. The present study demonstrates a largely under-recognized public health problem of a high unintentional injury risk for young people with ID, and identifies substantial and important risk factors for injury. Injury prevention programmes, with a particular focus on those with highly disturbed behaviours or epilepsy, are warranted in this population. PMID- 11851853 TI - Mothers with learning difficulties and their support networks. AB - Mothers with learning difficulties are thought to be among the most socially isolated parents in the community. A great deal of attention has been directed to assessing their parenting abilities and teaching parenting skills, but less has been given to the support that mothers may (or may not) receive from family, friends and the service system. The present paper investigates mothers' views about the types of support which they receive and from whom they receive it. Data were derived from 70 mothers who participated in interviews using a support interview guide designed to accommodate the mothers' cognitive difficulties. The primary purpose of the interview was to explore the quantity and composition of the mothers' support networks, the frequency of contact and geographical proximity of support people, and the type of support provided. Key findings include: the central place that family members have in these mothers' lives; the importance of service providers as sources of information and advice; and the relative absence of friends and neighbours. Briefly, mothers living alone have service-centred networks, mothers living with a partner have family-centred networks with relatively dispersed family ties, and mothers living in a parent/parent-figure household have local, family-centred networks. The overall conclusion to be drawn from the present results is that these mothers do not live in a social vacuum, but many are socially isolated. The finding that so few mothers could identify supportive ties with friends and neighbours suggests that these mothers are isolated from their local communities and are potentially vulnerable if a breakdown occurs in the support provided by their families. The need for service providers to be more actively involved in linking mothers to their communities is discussed. PMID- 11851854 TI - Communication profiles of individuals with Down's syndrome, Angelman syndrome and pervasive developmental disorder. AB - The communication profiles of individuals with Down's syndrome (DS), Angelman syndrome (AS) and pervasive developmental disorder (PDD) were investigated and contrasted. Seventy-seven individuals participated in the study. A within-group analysis revealed that those with DS performed better on tacting or labelling and echoing than on manding or requesting. No other effects were found, apart from an absence of echoing in those with AS, a result that is hardly surprising. A between-groups analysis revealed no differences between the aetiological groups in terms of their use of mands or requests and tacts. Individuals with DS and PDD did not differ in their scores on echoic functioning. The implications of these findings for the study of behavioural phenotypes and for communication intervention are discussed. PMID- 11851855 TI - Prader-Willi syndrome: new insights in the behavioural and psychiatric spectrum. AB - Prader-Willi syndrome (PWS) is a genetic disorder caused by the loss of the paternal contribution of the proximal part (15q11-q13) of the long arm of chromosome 15 (i.e. deletion, disomy and imprinting mutation). The syndrome is associated with distinct physical dysmorphism, as well as with specific behavioural and psychopathological characteristics. Psychiatric symptoms in adolescence and adulthood have been described, including acute cycloid psychosis, and obsessive compulsive, bipolar and pervasive developmental disorders. At the Centre for Human Genetics in Leuven, Belgium, 53 individuals (31 children and adolescents, and 22 adults) have been followed up for 15 years by a special multidisciplinary team. Attention was given to their medical, cognitive, behavioural and emotional development, and the evolution of psychiatric disorders in adolescence and adulthood. This study describes the psychiatric problems in four patients diagnosed with acute cycloid psychosis and traces their development from infancy to adolescence. Four other individuals needed psychiatric evaluation and treatment, and could be diagnosed as having unspecified bipolar disorder, also termed unstable mood disorder. Both groups were compared, and significant differences in early development and later evolution into adulthood were noted. The individuals with PWS who later developed psychotic episodes were described as active and extrovert toddlers, and showed autistic behaviour during their primary school education. Their intellectual functioning was in the moderate to severely retarded range. The individuals with PWS who later developed an unstable mood disorder were described as rather passive and introvert toddlers, and they presented less disturbed behaviour during their primary school education. The intellectual functioning of these subjects was in the normal to borderline range. PMID- 11851856 TI - Environmental, medical, behavioural and disability factors associated with Helicobacter pylori infection in adults with intellectual disability. AB - In institutionalized adults with intellectual disability (ID), Helicobacter pylori infection occurs at approximately twice the rate it appears in the general population, and it may be responsible for the twofold higher rates of peptic ulcer disease and gastric cancer in this population. Medical, behavioural and additional environmental factors, as well as level of ID, may be related to the risk of infection with H. pylori. One hundred and sixty-eight adults with ID who were currently, had previously been or had never been institutionalized underwent a biopsychosocial evaluation. This included assessment of: level of ID using the Adaptive Behaviour Scale (ABS) Part I; levels of maladaptive behaviour using the ABS Part II; demographic, medical and environmental factors; as well as H. pylori tests using serology and faecal antigen. The overall rates of past or current infection with H. pylori in institutionalized and previously institutionalized participants were about twice that of the overall group of never institutionalized participants, i.e. 87% and 79% compared to 44%, respectively (P < 0.001). The rates of H. pylori infection appeared to increase with age in the never-institutionalized group, but were consistently high across all ages in the other groups. The rate of infection was higher in those institutionalized for more than 5 years (95% versus 76%, P=0.02), in those with flatmates with excessive oral secretions (65% versus 21%, P < 0.001) or faecal incontinence (67% versus 27%, P < 0.001), and in those with more chronic illness and medications. All mean domain scores of the ABS Part I (Intellectual Disability) were significantly lower (indicating more severe ID) in the group currently infected with H. pylori compared to their non-infected counterparts. The majority of mean domain scores of the ABS Part II (Behaviour) were also worse, with half of these score differences reaching statistical significance in the currently infected group. The presence of alarm symptoms (e.g. vomiting, weight loss, haematemesis and melena), iron deficiency and body mass index were not significantly different in currently infected subjects. Adults with ID appear to be particularly at risk of infection with H. pylori. Environmental associations with infection include past or current institutionalization, a longer period of institutionalization, living with flatmates with excessive oral secretions and faecal incontinence. Medical associations include chronic disease and more medications, but not alarm symptoms or body mass index. Demographic associations may include increasing age in never-institutionalized adults, but no age effect in currently or previously institutionalized individuals. Psychosocial associations include more severe ID and maladaptive behaviour with current infection. PMID- 11851857 TI - Psychometric properties of the revised Developmental Behaviour Checklist scales in Dutch children with intellectual disability. AB - The present study assessed the reliability and validity of the revised scales of the Developmental Behaviour Checklist (DBC) in a Dutch sample of children with intellectual disability (ID). The psychometric properties of the parent and teacher versions of the DBC were assessed in various subsamples derived from a sample of 1057 Dutch children (age range=6-18 years) with ID or borderline intellectual functioning. Good test-retest reliability was shown both for the parent and teacher versions. Moderate inter-parent agreement and high one-year stability was found for the scale scores. Construct validity was satisfactory, although limited by high informant variance. The DBC scales showed good criterion related validity, as indicated by significant mean differences between referred and non-referred children, and between children with and without a corresponding DSM-IV diagnosis. The reliability and validity of the revised DBC scales are satisfactory, and the checklist is recommended for clinical and research purposes. PMID- 11851858 TI - Quantitative comparison of radial cell columns in children with Down's syndrome and controls. AB - No one has examined the configuration of the minicolumns in Down's syndrome (DS) brains even though these are a basic functional unit of the cortex. In the present study, the authors used computerized imaging to examine minicolumns in the posterior superior temporal gyrus in both the brains of patients with DS and normal controls. They compared the brains of children aged 4 and 6 years with those of adults for both people with DS and the normal population. Columns in the brains of two DS children aged 4 and 6 years were almost the same size as those of the adults with DS. The neuropil space in the periphery of the columns was also considerably wider. In contrast, minicolumns in aged-matched control children were smaller, both relatively and absolutely, when compared to the mean size of adult columns. The size of the minicolumns in the normal children apparently corresponded to the overall brain size, whereas the large columns in children with DS appeared to be independent of brain size, at least in area Tpt. This seems to reflect a rapid ageing process that is striking when compared to normal controls. Columns in adults with DS were large and less cell dense, while brain volumes were significantly smaller than in controls. This combination suggests reduced neuronal complexity based on a decrease in processing units, which supports previous findings of decreased cell numbers and synaptic diminution in DS brains. PMID- 11851859 TI - Ethical issues involved in the implementation of a differential reinforcement of inappropriate behaviour programme for the treatment of social isolation and ritualistic behaviour in an individual with intellectual disabilities. AB - The present case study describes an adult male with a 28-year history of institutional care underpinned by a moderate intellectual disability (ID), and ritualistic behaviour congruent with features of autism or obsessive-compulsive disorder. The subject's ID was recognized early in life and he was provided with a special educational placement at 6 years of age, although his increasingly disturbed behaviour had resulted in several psychiatric admissions by early adolescence, and by the age of 20, his presentation necessitated long-term secure psychiatric care. The present report describes the application and ethical issues pertaining to a differential reinforcement of inappropriate behaviour (DRI) programme for the reduction of ritualistic behaviour and social isolation, the latter being a direct consequence of the former, one year after the subject was admitted to a medium-secure assessment and treatment residential facility. The results suggest that, although the treatment was successful in targeting ritualization and reducing social isolation, ethical concerns extended to care staff concerns about the reinforcer adopted for the programme and the lack of informed consent from the subject. The former resulted in inconsistent application of the programme and its eventual termination. A follow-up assessment of the subject's extent of social isolation and ritualistic behaviour suggested a return to a pre-DRI level of morbidity. Limitations and suggestions for the future treatment of such cases are outlined. PMID- 11851860 TI - The genetical history of humans and the great apes. AB - When and where did modern humans evolve? How did our ancestors spread over the world? Traditionally, answers to questions such as these have been sought in historical, archaeological, and fossil records. However, increasingly genetic data provide information about the evolution of our species. In this review, we focus on the comparison of the variation in the human gene pool to that of our closest evolutionary relatives, the great apes, because this provides a relevant perspective on human genetical evolution. For instance, comparisons to the great apes show that humans are unique in having little genetic variation as well as little genetic structure in their gene pool. Furthermore, genetic data indicate that humans, but not the great apes, have experienced a period of dramatic growth in their early history. PMID- 11851861 TI - Serum free fatty acid pattern and risk of myocardial infarction: a case-control study. AB - OBJECTIVES: To investigate the association between composition of serum free fatty acid (FFA) fraction and risk of a first myocardial infarction (MI). DESIGN: A case-control design. SETTING: The patients were recruited from Ulleval Hospital in Oslo and Ostfold Central Hospital in Fredrikstad and Sarpsborg, Norway. SUBJECTS: A total of 103 patients with first MI and 104 population controls, both men and postmenopausal women, age 45-75 years. RESULTS: The mean molar percentage content of docosahexaenoic (DHA), eicosapentaenoic (EPA), stearic and myristic acid in the serum FFA fraction was significantly lower in cases than in controls, whereas that of oleic and linoleic acid was higher in cases. Increased percentage content of total very long-chain omega-3 fatty acids (VLC n-3) in serum FFA was associated with decreased risk of MI. Multivariate odds ratio (OR), adjusted for age, sex, waist-hip ratio, smoking, family history of coronary heart disease (CHD) and years of education was 0.20 (95% CI 0.06-0.63) for the highest vs. lowest quartile. Also increased content of stearic acid was associated with decreased risk. Multivariate OR adjusted as above was 0.38 (95% CI 0.14-1.04) for the highest versus lowest quartile. After adjustment for oleic acid, however, the inverse linear trend was no longer significant. CONCLUSIONS: The percentage content of VLC n-3 as well as of stearic acid in serum FFA was inversely associated with risk of myocardial infarction. That of VLC n-3 may reflect diet, but additionally these free fatty acids might in some way be related to the pathogenetic process and not only reflect their content in adipose tissue. PMID- 11851862 TI - Antineutrophil cytoplasmatic antibodies in patients with premature atherosclerosis: prevalence and association with risk factors. AB - OBJECTIVES: Autoimmunity is suggested to play a role in premature atherosclerosis. Antineutrophil cytoplasmatic antibodies (ANCA) are a group of autoantibodies found in several inflammatory disorders in which they supposedly amplify the inflammatory process. In this study the hypothesis is tested that ANCA play a role in premature atherosclerosis. DESIGN: Cross-sectional study followed by nested case-control study. In a total of 286 consecutive patients with premature atherosclerosis (age < 55 years) ANCA were tested. Within the same cohort, a nested case-control study in 16 ANCA-positive patients and 32 ANCA negative controls matched for sex, and site of atherosclerosis, was executed. SETTING: University hospital outpatient clinic for lipids and premature atherosclerosis. SUBJECTS: A total of 286 consecutive patients with premature atherosclerosis (age < 55 years). RESULTS: Prevalence of ANCA was 5.6% (16 of 286). All cases had perinuclear ANCA (pANCA); no cytoplasmatic ANCA was found. Mean age was 42 +/- 7 in the ANCA-positive vs. 42 +/- 9 years in the ANCA negative group (P=ns). More female parents were ANCA-positive (8M/8F vs. 200M/70F, P=0.03). Patients with ANCA had more often peripheral vascular disease (37.5 vs. 15.2%, P=0.03). In the case-control study levels of Lp(a) were higher (43.8 vs. 15.6% >300 mg x L(-1), P=0.05), whereas levels of HDL-c were lower in ANCA-positive patients (0.84 +/- 0.26 vs. 1.06 +/- 0.27 mmol x L(-1), P=0.01). Markers of inflammation, C-reactive protein (CRP) and serum amyloid A (SAA), did not differ, nor did antibodies against oxidized-LDL and malondialdehyde (MDA) LDL, markers for the extent of atherosclerosis. CONCLUSIONS: Our results suggest that ANCA do not appear to play a major role in premature atherosclerosis as there was no increased prevalence of the autoantibody. Moreover, no differences in the incidence of classical cardiovascular risk factors nor in serum levels of markers of inflammation were found between the ANCA-positive group as compared with the ANCA-negative group. PMID- 11851863 TI - Visceral fat and psychosocial stress in identical twins discordant for obesity. AB - OBJECTIVES: Accumulation of intra-abdominal fat has been suggested, but not yet proved, to be basically as a result of chronic psychosocial stress causing arousal of hypothalamic-pituitary-adrenal cortex axis. Our objectives were to study the association between psychosocial stress, obesity and body fat distribution when genetic factors are identical. DESIGN: Monozygotic twins discordant for obesity were examined in an in-patient setting. SUBJECTS: Adult monozygotic twin pairs (12 female, 8 male) with an average intrapair difference of 17 kg in body weight. They were divided into two groups: in group A the visceral fat area of the obese cotwin was higher and, in group B, lower than the gender-specific median value. MAIN MEASURES: Hormonal, physiological and psychological distress indicators, and sleep measures. RESULTS: Daily urinary cortisol and noradrenaline excretion were higher in the obese cotwins when compared with the nonobese cotwins in group A but not in group B (P=0.026 and 0.020 when intrapair differences were compared between groups A and B, respectively). In serum cortisol, ACTH and CBG concentrations a similar trend was not statistically significant. In group A, the obese cotwins consumed almost 2.5 times as much alcohol as their lean cotwins, whilst in group B the situation was the opposite. The mean amount of active sleep was significantly higher and that of quiet sleep significantly lower in the obese than the lean cotwins only in group A. Intrapair differences in emotional reactions indicating distress and lack of subjective energy were seen only in group A. CONCLUSION: When genetic factors are identical, visceral fat accumulation, rather than obesity in general, is associated with increased psychosocial stress and concomitant hormonal changes. PMID- 11851864 TI - Costs for screening, intervention and hospital treatment generated by the Malmo Preventive Project: a large-scale community screening programme. AB - OBJECTIVES: The purpose of this study was to estimate retrospectively the costs of health care resources used in the Malmo Preventive Project, Sweden and estimate the costs of in-patient care that were avoided because of early intervention. SETTING AND SUBJECTS: A large-scale community intervention programme was conducted from 1974 to 1992 in Malmo, Sweden with the aim of reducing morbidity and mortality of cardiovascular diseases (CVD), alcohol related illnesses, and breast cancer. Between 1974 and 1992, 33 336 male and female subjects were screened for hypertension, hyperlipidaemia, type-2 diabetes and alcohol abuse. Intervention programmes that included life-style modifications, follow-up visits with physicians and nurses and drug therapy were offered to about 25% of screened subjects. METHODS: Recruitment costs were generated through out the screening period. Intervention costs were estimated for 5 years after screening. Excess in-patient care costs were estimated by subtracting hospital consumption for an unscreened, matched cohort from that of the screened cohort over follow-up periods of 13-19 years. Intervention and excess in-patient care costs were estimated until 1996. RESULTS: The net expenditures for recruitment and intervention was SEK253 million and saved costs for in-patient care of SEK143 millions (1998 prices). Considering the opportunity cost of the resources used in the study, the net cost rises to about SEK200 millions. CONCLUSIONS: The results suggest that only part of the intervention costs were offset by reduction in future morbidity health care costs. This is in line with results from prospective analyses of other primary prevention programmes. PMID- 11851865 TI - Exercise-induced symptomatic ischaemia predicts a poor long-term prognosis after acute myocardial infarction. AB - STUDY OBJECTIVES: To evaluate the long-term prognostic significance of symptomatic ischaemia during exercise testing performed 3 weeks after acute myocardial infarction (AMI). DESIGN: A prospective study with long-term follow up. SETTING: A Cardiac Rehabilitation Clinic in a University Hospital. SUBJECTS: A total of 446 patients were allowed to perform exercise testing 3 weeks after AMI and followed for 72 +/- 20 months. MEASUREMENTS AND RESULTS: Patients were divided into three groups according to whether they had no ECG evidence of ischaemia during exercise testing (334 patients), silent ischaemia (90 patients) or symptomatic ischaemia (22 patients). Cardiac death was significantly more frequent in patients with symptomatic ischaemia when compared with silent ischaemia (31.8% vs. 7.8%, P < 0.01) or when compared with no ischaemia (31.8% vs. 10.2%, P < 0.01). The three groups had a low cardiac mortality during the first 48 months of follow-up. The prognosis of patients with symptomatic ischaemia worsens markedly thereafter. The results of exercise testing did not predict recurrence of myocardial infarction. Coronary revascularization was performed in 34.4% of those without ischaemia, 47.8% of those with silent ischaemia and 45.5% of those with symptomatic ischaemia (P < 0.01). CONCLUSIONS: Patients with symptomatic ischaemia have a good prognosis during the first 4 years of follow-up. Their prognosis worsens thereafter as opposed to patients with or without silent ischaemia. This high-risk group of patients with symptomatic ischaemia deserves optimal management including revascularization when appropriate. PMID- 11851866 TI - Interleukin-1 receptor antagonist is detectable in human carotid artery plaques and is related to triglyceride levels and Chlamydia pneumoniae IgA antibodies. AB - OBJECTIVES: To investigate whether the interleukin-1 receptor antagonist (Il-1ra) and interleukin-1beta (Il-1beta) can be detected in human carotid artery tissue, and whether their presence is related to evidence of Chlamydia pneumoniae infection, risk factors for atherosclerosis, and clinical data. SETTING: Departments of Vascular Diseases and Surgical Pathophysiology, University Hospital, Malmo, Sweden. SUBJECTS: A total of 66 patients undergoing carotid endarterectomy (median age 74, range 53-89 years, 26 women). Il-1beta and Il-1ra were studied in carotid artery plaques and in Il-1ra in serum. RESULTS: Interleukin-1 receptor antagonist was detected in mononuclear cells in plaques from 37/66 (56%) patients. Patients with Il-1ra in plaques showed higher [2.04 (1.70-3.14) mmol x L(-1) vs. 1.69 (1.09-1.99) mmol x L(-1); P < 0.05] serum(s )triglyceride(tg) levels, and a higher frequency of IgA seropositivity for C. pneumoniae (76% vs. 52%; P < 0.05) than those without. S-Il-1ra levels correlated with s-tg levels (r=0.38; P=0.047). There were no differences between patients with and without Il-1ra in plaques concerning s-Il-1ra, blood(b-)haemoglobin or leucocyte count, s-cholesterol, b-glucose, blood pressure, IgG seropositivity for C. pneumoniae, prevalence of neurological symptoms preceding operation, smoking, or diabetes mellitus. There were no differences in frequency of Il-1ra in plaques or in s-Il-1ra levels between patients with symptomatic and asymptomatic stenosis, between smokers and nonsmokers, or between diabetic and nondiabetic patients. Il-1beta was not detected in plaques in the current study. CONCLUSION: Interleukin-1 receptor antagonist can be detected in human atherosclerotic carotid artery plaques, and is related to s-triglyceride levels and IgA seropositivity for C. pneumoniae, but not to prevalence of neurological symptoms related to embolization. PMID- 11851867 TI - Oxidative stress and antioxidant status in type 1 diabetes mellitus. AB - OBJECTIVES: To test the hypothesis that type 1 diabetes is associated with increased oxidative stress and/or antioxidant status by investigating concentrations of 8-iso-prostaglandin F2alpha (8-iso-PGF2alpha) in urine and plasma and malondialdehyde (MDA) in plasma as indicators of lipid peroxidation in vivo, and antioxidant status in diabetic subjects compared with healthy control subjects. DESIGN AND SUBJECTS: Thirty-eight subjects with type 1 diabetes mellitus and 41 healthy age- and sex-matched control subjects were included in the study. Blood and urine samples were obtained and analysed for 8-iso-PGF2alpha with a newly developed radioimmunoassay, as well as for MDA, total antioxidant capacity (TAOC) and serum tocopherol levels. RESULTS: None of the variables of lipid peroxidation showed any significant difference between the two groups. Similarly, there were no significant correlations between the levels of 8-iso PGF2alpha or MDA, and degree of glycemic control (HbA1c). Total antioxidant capacity in plasma was 16% lower amongst the subjects with type 1 diabetes than in the control group (P < 0.0005). Lipid corrected levels of alpha-tocopherol in serum were significantly increased in type 1 diabetic subjects (P < 0.05), as were gamma-tocopherol levels (P < 0.005). CONCLUSIONS: In spite of lower total antioxidant defence, our results do not support the oxidative stress hypothesis for type 1 diabetes mellitus. The higher tocopherol levels suggest that no vitamin E supplementation is necessary for subjects with type 1 diabetes mellitus. PMID- 11851868 TI - Both cerivastatin and fenofibrate improve arterial vasoreactivity in patients with combined hyperlipidaemia. AB - OBJECTIVES: The aim of this study was to compare the effects of cerivastatin and fenofibrate on endothelium dependent and independent arterial dilation. DESIGN: In a prospective, double blind study, 38 overweight, nonsmoking, males aged between 40 and 60 years with combined hyperlipidaemia were randomized and, after 6 weeks run-in phase with American Heart Association step I diet treatment, submitted to 12 weeks' treatment either with fenofibrate (250 mg daily) or cerivastatin. Cerivastatin was given in a daily dose of 0.2 mg for 6 weeks and was increased to 0.4 mg daily, if the LDL-C did not decrease below 3.0 mmol x L( 1). Flow-mediated (endothelium-dependent) dilation (FMD) and nitroglycerin induced (endothelium-independent) [gliceryltrinitrate (GTN)] dilation of brachial artery were measured using high resolution ultrasound. RESULTS: The FMD increased from 3.4 +/- 3.3 to 9.3 +/- 2.4% (P < 0.001) in the cerivastatin group, and from 3.3 +/- 2.8 to 6.5 +/- 3.1% (P < 0.001) in the fenofibrate group, the improvement being significantly better after cerivastatin (P=0.006). GTN increased from 11.5 +/- 4.1 to 16.2 +/- 3.5% (P < 0.01) and from 11.1 +/- 2.5 to 16.0 +/- 2.9% (P < 0.01), respectively, with no difference between the groups. Cerivastatin reduced total cholesterol by 24%, LDL-cholesterol by 31%, triglycerides by 24%, ox-LDL by 29% and increased HDL-cholesterol by 5%, whilst, after fenofibrate, these changes were -15, -13, -41, -17 and 18%, respectively. Only the decrease of LDL-C turned out to be an independent predictor the FMD improvement. The improvement in GTN induced dilation did not correlate with the changes in blood lipids. CONCLUSIONS: Both cerivastatin and fenofibrate lead to an improvement of endothelium-dependent and endothelium-independent dilation of brachial artery in overweight patients with combined hyperlipidaemia and no other atherosclerotic risk factors. The effects on FMD were greater in subjects receiving cerivastatin than in subjects receiving fenofibrate, but the effects on GTN were equal in both groups. PMID- 11851869 TI - Modulation of BMP signaling by noggin is required for induction of the secondary (nontylotrich) hair follicles. AB - Increasing evidence suggests that morphogenesis of the distinct developmental structures derived from the same organ-committed epithelium is controlled by differential mechanisms. As was recently shown in mice with mutations in the downless (dL) gene, induction of primary or tylotrich hair follicles is strikingly dependent of signaling through the Tnf receptor homologue, Edar. Here, we show that dorsal skin of murine embryos with constitutive deletion of the BMP2/4 antagonist noggin, after transplantation into SCID mice, is characterized by the lack of induction of secondary hair follicles, and by the arrest of primary hair follicle development prior to hair shaft formation. The loss of noggin activity was associated with failure to express genes that specify hair follicle cell fates in the epidermis (Lef-1, beta-catenin, Shh) and dermal papilla (p75 kDa neurotrophin receptor, alkaline phosphatase). This suggests that regulation of BMP2/4 signaling by noggin is essential for the induction of secondary hair follicles, as well as for advanced stages of development in primary hair follicles. PMID- 11851871 TI - Contrasting expression patterns of CCAAT/enhancer-binding protein transcription factors in the hair follicle and at different stages of the hair growth cycle. AB - Hair follicles undergo repeated cycles of growth and regression, throughout the entire life of the organism. These dynamic changes require closely co-ordinated regulation of gene expression. The CCAAT/enhancer-binding proteins are a family of basic region/leucine zipper transcription factors that regulate gene transcription in various tissues. They have been implicated in epidermal differentiation and may therefore play an important role in the hair follicle. We have investigated the localization of four members of this family--CCAAT/enhancer binding protein-alpha, -beta, and -delta, and Gadd153--in both human and murine hair follicles by immunohistochemistry. Furthermore, we examined CCAAT/enhancer binding protein-alpha, -beta, and -delta immunoreactivity at different stages of the depilation-induced murine hair growth cycle. Distinct immunoreactivity patterns for CCAAT/enhancer-binding protein-alpha, -beta, and -delta, and Gadd153 were observed in the outer root sheath, sebaceous gland, dermal papilla, and connective tissue sheath of human anagen hair follicles. In murine follicles, CCAAT/enhancer-binding protein-alpha was expressed in the outer root sheath, sebaceous gland, and dermal papilla, whereas CCAAT/enhancer-binding protein-beta expression was confined to the matrix, sebaceous gland, and inner and outer root sheaths. Both CCAAT/enhancer-binding protein-alpha and -beta were upregulated during anagen, then downregulated in catagen follicles. In contrast, CCAAT/enhancer-binding protein-delta showed no hair cycle-dependent variation in immunoreactivity. These data suggests that the expression of CCAAT/enhancer binding protein-alpha and -beta may, in turn, play a part in regulating hair cycle-dependent gene expression. Moreover, as CCAAT/enhancer-binding protein alpha, -beta, and -delta are crucial in the regulation of adipocyte differentiation and lipid metabolism, their expression in sebocytes suggests they may also play a similar role in differentiation and lipid metabolism of the sebaceous gland. PMID- 11851870 TI - Lack of the vitamin D receptor is associated with reduced epidermal differentiation and hair follicle growth. AB - The active vitamin D metabolite, 1,25-dihydroxyvitamin D, acting through the vitamin D receptor, regulates the expression of genes in a variety of vitamin D responsive tissues, including the epidermis. To investigate the role of the vitamin D receptor in mediating epidermal differentiation, we examined the histomorphology and expression of differentiation markers in the epidermis of vitamin D receptor knockout mice generated by gene targeting. The homozygous knockout mouse displayed a phenotype that closely resembles vitamin D-dependent rickets type II in humans, including the development of rickets and alopecia. Hair loss developed by 3 mo after birth and gradually led to nearly total hair loss by 8 mo. Histologic analysis of the skin of homozygous knockout mice revealed dilation of the hair follicles with the formation of dermal cysts starting at the age of 3 wk. These cysts increased in size and number with age. Epidermal differentiation markers, including involucrin, profilaggrin, and loricrin, detected by immunostaining and in situ hybridization, showed decreased expression levels in homozygous knockout mice from birth until 3 wk, preceding the morphologic changes observed in the hair follicles. Keratin 10 levels, however, were not reduced. At the ultrastructural level, homozygous knockout mice showed increased numbers of small dense granules in the granular layer with few or no surrounding keratin bundles and a loss of keratohyalin granules. Thus, both the interfollicular epidermis and the hair follicle appear to require the vitamin D receptor for normal differentiation. The temporal abnormalities between the two processes reflect the apparent lack of requirement for the vitamin D receptor during the anagen phase of the first (developmental) hair cycle, but with earlier effects on the terminal differentiation of the interfollicular epidermis. PMID- 11851872 TI - Oxysterol stimulation of epidermal differentiation is mediated by liver X receptor-beta in murine epidermis. AB - Liver X receptor-alpha and -beta are members of the nuclear hormone receptor superfamily that heterodimerize with retinoid X receptor and are activated by oxysterols. In recent studies we found that treatment of cultured human keratinocytes with oxysterolstimulated differentiation, as demonstrated by increased expression of involucrin and transglutaminase, and inhibited proliferation. The aims of this study were to determine: (i) whether oxysterols applied topically to the skin of mice induce differentiation in normal epidermis; (ii) whether this effect is mediated via liver X receptor-alpha and/or liver X receptor-beta; and (iii) whether oxysterols normalize epidermal morphology in an animal model of epidermal hyperplasia. Topical treatment of normal hairless mice with 22(R)-hydroxycholesterol or 24(S),25-epoxycholesterol resulted in a decrease in epidermal thickness and a decrease in keratinocyte proliferation assayed by proliferating cell nuclear antigen staining. Moreover, oxysterol treatment increased the levels of involucrin, loricrin, and profilaggrin protein and mRNA in the epidermis, indicating that oxysterols stimulate epidermal differentiation. Additionally, topical oxysterol pretreatment improved permeability barrier homeostasis. Whereas liver X receptor-alpha-/- mice revealed no alterations in epidermal differentiation, the epidermis was thinner in liver X receptor-beta-/- mice than in wild-type mice, with a reduced number of proliferating cell nuclear antigen positive cells and a modest reduction in the expression of differentiation markers. Topical oxysterol treatment induced differentiation in liver X receptor-alpha-/- mice whereas in liver X receptor-beta-/- mice there was no increase in the expression of differentiation markers. Whereas both liver X receptor-alpha and liver X receptor-beta are expressed in cultured human keratinocytes and in fetal rat skin, only liver X receptor-beta was observed on northern blotting in adult mouse epidermis. Finally, treatment of hyperproliferative epidermis with oxysterols restored epidermal homeostasis. These studies demonstrate that epidermal differentiation is regulated by liver X receptor-beta and that oxysterols, acting via liver X receptor-beta, can induce differentiation and inhibit proliferation in vivo. The ability of oxysterols to reverse epidermal hyperplasia suggests that these agents could be beneficial for the treatment of skin disorders associated with hyperproliferation and/or altered differentiation. PMID- 11851873 TI - All-trans retinoic acid induces differentiation and apoptosis of murine melanocyte precursors with induction of the microphthalmia-associated transcription factor. AB - The effects of all-trans retinoic acid on the differentiation and proliferation of immature melanocyte precursors were studied. NCC-melb4 cells are an immortal cloned cell line established from mouse neural crest cells using a single-cell cloning method. These cells were positive for tyrosinase-related protein 1, tyrosinase-related protein 2 and KIT, but were negative for tyrosinase and had no dihydroxyphenylalanine reaction. They contained only stage I melanosomes without any melanosomes in more advanced stages. After treatment with all-trans retinoic acid, many of the cells became tyrosinase- and dihydroxyphenylalanine-reaction positive, changed from polygonal to dendritic in shape, and had stage III to IV melanosomes. These findings indicate that treatment with all-trans retinoic acid induced the differentiation of NCC-melb4 cells. Reverse transcription polymerase chain reaction analysis revealed a marked increase in expression of microphthalmia-associated transcription factor mRNA after all-trans retinoic acid treatment, suggesting that microphthalmia-associated transcription factor may be the key molecule in this event. Enhanced expression of protein kinase Calpha following treatment with all-trans retinoic acid was also demonstrated. The proliferation of NCC-melb4 cells was inhibited by all-trans retinoic acid in a dose-dependent manner. Increased apoptosis after all-trans retinoic acid treatment was observed by electron microscopy, the TUNEL method, DNA fragmentation assay, and flow cytometry. All-trans retinoic acid upregulated caspase-3 and downregulated bcl-2. Electron microscopy showed that apoptotic cells contained melanosomes of advanced stages, suggesting that mature melanocytes may tend to undergo apoptosis after all-trans retinoic acid treatment. This study provides important clues towards understanding the roles and working mechanisms of retinoic acids in melanocyte development and melanogenesis. PMID- 11851874 TI - Hyaluronan synthase 3 regulates hyaluronan synthesis in cultured human keratinocytes. AB - Three human hyaluronan synthase genes (HAS1, HAS2, and HAS3) have been cloned, but the functional differences between these HAS genes remains obscure. The purpose of this study was to examine which of the HAS genes are selectively regulated in epidermis. We examined the relation of changes between hyaluronan production and HAS gene expression when cytokines were added to cultured human keratinocytes. Interferon-gamma increased hyaluronan production whereas transforming growth factor beta decreased it. Both cytokines affected preferentially high-molecular-mass (> 106 Da) hyaluronan production. Consistent with the change in hyaluronan synthesis, we found that interferon-gamma markedly upregulated HAS3 mRNA whereas transforming growth factor beta downregulated HAS3 transcript levels. The expression of HAS1 mRNA was not significantly affected by either cytokine, and HAS2 mRNA expression was undetectable under either basal or cytokine-stimulated conditions by northern blot using total RNA. Furthermore, in situ mRNA hybridization showed that mouse epidermal keratinocytes abundantly expressed HAS3 mRNA from the basal to the granular cell layers, suggesting that HAS3 functions in epidermis. These findings suggest that HAS3 gene expression plays a crucial role in the regulation of hyaluronan synthesis in the epidermis. PMID- 11851875 TI - Human leukocyte elastase induces keratinocyte proliferation in vitro and in vivo. AB - Neutrophil infiltration and epidermal hyperproliferation are major histopathologic changes observed in psoriasis. Neutrophils contain human leukocyte elastase, which is thought to be released during neutrophil infiltration of the epidermis. As active human leukocyte elastase is known to be present in psoriatic lesions we were interested whether human leukocyte elastase induces hyperproliferation in keratinocytes in vitro and in vivo. In the cultured murine keratinocyte cell line PAM-212 concentrations of human leukocyte elastase from 1 to 30 nM induced significant proliferation as determined by 5-bromo-2' deoxy-uridine-incorporation. Daily topical application of 0.043-434.8 pmol human leukocyte elastase per cm2 skin on hairless mice induced a concentration dependent epidermal hyperproliferation and an increase in 5-bromo-2'-deoxy uridine incorporation of up to 5-fold in basal keratinocytes within 3 d. Hyperproliferation resulted in a up to 2-fold increase of keratinocyte layers. Histologic analysis revealed marked vasodilatation but no inflammatory infiltrate. Application of porcine pancreatic elastase (3-300 pmol per cm2 skin) resulted in similar epidermal changes as observed for human leukocyte elastase. Hyperproliferative effects of human leukocyte elastase in vitro and in vivo were abolished by the addition of elastase inhibitors, such as elafin, anti leukoprotease, and alpha1-protease inhibitor. In summary, human leukocyte elastase induces proliferation in murine keratinocytes in concentrations, which can be found on the skin surface of psoriatic lesions. These results may provide an explanation for the epidermal hyperproliferation observed in psoriasis. PMID- 11851876 TI - Matrix metalloproteinase inhibitor BB-3103 unlike the serine proteinase inhibitor aprotinin abrogates epidermal healing of human skin wounds ex vivo. AB - Several matrix metalloproteinases and serine proteinases are upregulated in migrating keratinocytes during cutaneous wound repair. Single cell culture studies indicate the necessity for matrix metalloproteinases but not for serine proteinases in keratinocyte locomotion. To account for epithelial-mesenchymal interactions, an ex vivo human skin wound model was used to investigate the contribution of matrix metalloproteinases and serine proteinases to wound healing by treatment with broad-spectrum inhibitors of matrix metalloproteinases (BB 3103) or serine proteinases (aprotinin). Human skin explants with circular 3 mm superficial defects were incubated in culture medium without (controls) or with the proteinase inhibitors for 7 d. BB-3103 abrogated epithelialization (p < 0.001), whereas aprotinin-treated wounds and controls were covered with new epithelium. Lack of epithelialization was unlikely due to cytotoxicity because the matrix metalloproteinase inhibitor did neither influence viability of cultured epidermal keratinocytes nor apoptosis in wounds. Involvement of specific matrix metalloproteinases in epithelialization was analyzed by gelatin zymography, western blotting, immunohistochemistry, and in situ hybridization. Wound healing was accompanied by active matrix metalloproteinase-1 and increased active matrix metalloproteinase-2 but irrespectively of active matrix metalloproteinase-9. BB-3103 blocked activation of matrix metalloproteinase-2 and matrix metalloproteinase-9 but not of matrix metalloproteinase-1. Active matrix metalloproteinase-2 localized solely to the dermis, whereas matrix metalloproteinase-9 was consistently found in new epithelium. Membrane-type 1 matrix metalloproteinase was undetectable in wound keratinocytes. BB-3103 and aprotinin reduced tumor necrosis factor-alpha in media but did not appreciably alter amounts of other soluble regulators of matrix metalloproteinases and epithelialization. Our findings demonstrate that keratinocyte migration is associated with active matrix metalloproteinase-2 but occurs independently of serine proteinases and active matrix metalloproteinase-9 in fibrin-deficient skin wound healing. PMID- 11851877 TI - Negative electric potential induces alteration of ion gradient and lamellar body secretion in the epidermis, and accelerates skin barrier recovery after barrier disruption. AB - Previous reports suggested that ion gradients of ions such as calcium and magnesium in the epidermis play a crucial part in skin barrier homeostasis. We hypothesized that external electric potential affects the ionic gradient and skin barrier homeostasis. We demonstrated here that application of a negative electric potential (0.50 V) on hairless mice skin accelerated the barrier recovery approximately 60.7% of the original level within 1 h compared with control (37.5%) after barrier disruption by acetone treatment. Even after the application of a negative potential, the skin showed accelerated repair for 6 h. On the contrary, the skin that was applied a positive potential for 1 h showed a significant delay in barrier recovery (25.3%) than the control. Ultrastructural studies by electron microscopy suggested that the extent of lamellar body exocytosis into the stratum corneum/stratum granulosum interface increased under a negative potential. Magnesium and calcium ion concentrations in the upper epidermis were relatively higher in the negative portion than in the portion where the positive potential was applied. Topical application of these ions on mice skin also accelerated the barrier recovery. These results suggest that the external electric potential affects the ionic gradients in the epidermis and also influences the skin barrier homeostasis. PMID- 11851878 TI - Increase of pro-opiomelanocortin mRNA prior to tyrosinase, tyrosinase-related protein 1, dopachrome tautomerase, Pmel-17/gp100, and P-protein mRNA in human skin after ultraviolet B irradiation. AB - In ultraviolet-induced tanning, the protein levels of various gene products critical for pigmentation (including tyrosinase and tyrosinase-related protein-1) are increased in response to ultraviolet B irradiation, but changes in mRNA levels of these factors have not been investigated in vivo. We have established an in situ hybridization technique to investigate mRNA levels of pro opiomelanocortin, tyrosinase, tyrosinase-related protein-1, dopachrome tautomerase, P-protein, Pmel-17/gp100, and microphthalmia-associated transcription factor, and have analyzed the changes in mRNA levels in the ultraviolet B-exposed skin in vivo. The right or left forearm of each volunteer was irradiated with ultraviolet B, and skin biopsies were obtained at 2 and 5 d postirradiation. mRNA level of pro- opiomelanocortin was increased 2 d after ultraviolet B irradiation, and returned to a near-basal level after 5 d, whereas the mRNA levels of tyrosinase, tyrosinase-related protein-1, dopachrome tautomerase, P-protein, and Pmel-17/gp100 showed some or no increase at 2 d, but were significantly increased 5 d after ultraviolet B irradiation. Microphthalmia associated transcription factor mRNA was slightly increased on days 2 and 5 after ultraviolet B irradiation. Our results suggest that the mechanism of the tanning response of human skin may involve the transcriptional regulation of certain pigmentary genes, and that pro-opiomelanocortin-derived melanocortins such as alpha-melanocyte-stimulating hormone and adrenocorticotropic hormone may play a part in regulating these genes in vivo. PMID- 11851879 TI - High-frequency microsatellite instability is associated with defective DNA mismatch repair in human melanoma. AB - Hereditary nonpolyposis colorectal cancers and a steadily increasing number of sporadic tumors display microsatellite instability. In colorectal tumors, high frequency microsatellite instability is strictly associated with inactivation of the DNA mismatch repair genes hMSH2, hMLH1, or hPMS2, whereas mutations in the mismatch repair gene hMSH6 have been identified in a subset of tumors with low frequency microsatellite instability. In addition to epithelial tumors of the colon, endometrium, and ovary, microsatellite instability has been reported to occur also in sporadic melanoma. The relationship between microsatellite instability and mismatch repair in melanoma cells, however, has not been investigated so far. In this study, we analyzed microsatellite instability, mismatch repair activity, and expression of the hMSH2, hMSH6, hMLH1, and hPMS2 proteins in five melanoma cell lines and in tumor specimens from which the cells were derived. Four cell lines displayed normal levels of mismatch repair activity and expressed all the mismatch repair proteins. The extracts of the fifth cell line lacked the hMLH1 and hPMS2 proteins, and were correspondingly deficient in the repair of DNA mismatches. This line displayed high-frequency microsatellite instability, whereas the four mismatch-repair-proficient cell lines displayed either no or low-frequency microsatellite instability. These findings could be confirmed in the tumor specimens, in that only the tumor that did not express hMLH1 and hPMS2 displayed high-frequency microsatellite instability. Our data are consistent with the hypothesis that in melanoma, similarly to epithelial tumors, only the high-frequency microsatellite instability phenotype is strictly dependent on a defective mismatch repair system. Further studies on a large series of tumor specimens are required to establish the frequency of mismatch repair loss in human melanoma. PMID- 11851880 TI - A site-specific plectin mutation causes dominant epidermolysis bullosa simplex Ogna: two identical de novo mutations. AB - Plectin is one of the largest and most versatile cytolinker proteins known. In basal keratinocytes it links the intermediate filament network to cell membrane associated hemidesmosomes. Several mutations in its gene have been identified that lead to the recessive disease epidermolysis bullosa with muscular dystrophy. We report here a mutation that leads to a dominant form of the disease, epidermolysis bullosa simplex Ogna. We found that the epidermolysis bullosa simplex Ogna phenotype is due to a site-specific missense mutation within plectin's rod domain. Further, we show that epidermolysis bullosa simplex Ogna is not restricted to a single Norwegian kindred as previously believed. A German family with the phenotypic hallmarks of epidermolysis bullosa simplex Ogna was found to carry an identical de novo mutation. These two mutations arose about 200 y apart in time. Consistent with the absence of muscular symptoms in these patients, muscle biopsies from several epidermolysis bullosa simplex Ogna members of the Norwegian kindred showed normal staining patterns using antibodies to plectin. Skin changes in epidermolysis bullosa simplex Ogna patients are documented on the ultrastructural level. PMID- 11851881 TI - Topical peroxisome proliferator activated receptor-alpha activators reduce inflammation in irritant and allergic contact dermatitis models. AB - Activators of peroxisome proliferator activated receptor-alpha, a nuclear hormone receptor that heterodimerizes with retinoid X receptor, stimulate epidermal differentiation and inhibit proliferation. Here we determined the anti inflammatory effects of peroxisome proliferator activated receptor-alpha agonists in models of irritant and allergic contact dermatitis produced in mouse ears by topical treatment with 12-O-tetradecanoylphorbol-13-acetate and oxazalone, respectively. As expected, 12-O-tetradecanoylphorbol-13-acetate treatment resulted in a marked increase in the thickness and weight of the ears and provoked an inflammatory cell infiltrate in the dermis. Topical treatment with three different peroxisome proliferator activated receptor-alpha agonists, clofibrate, WY 14643, or linoleic acid, 45 min and 4 h after 12-O tetradecanoylphorbol-13-acetate application, resulted in a marked decrease in ear thickness and weight and a reduction in the number of inflammatory cells in the dermis. The reduction in inflammation by these peroxisome proliferator activated receptor-alpha agonists was of similar magnitude to that seen with a potent topical glucocorticoid, clobetasol. In contrast, stearic acid, a free fatty acid that does not activate peroxisome proliferator activated receptor-alpha, had no effect on the 12-O-tetradecanoylphorbol-13-acetate-induced inflammation. Moreover, clofibrate did not significantly alter ear thickness following 12-O tetradecanoylphorbol-13-acetate treatment in peroxisome proliferator activated receptor-alpha-/- mice, indicating that the anti-inflammatory effect is mediated by peroxisome proliferator activated receptor-alpha. As tumor necrosis factor alpha and interleukin-1alpha are major mediators of cutaneous inflammation we next used immunohistochemistry to determine whether the peroxisome proliferator activated receptor-alpha agonists reduce the levels of these cytokines in 12-O tetradecanoylphorbol-13-acetate-treated skin. 12-O-tetradecanoylphorbol-13 acetate treatment resulted in an increase in tumor necrosis factor and interleukin-1alpha staining in the epidermis that was reduced by clofibrate treatment. Finally, clofibrate treatment also reduced ear thickness and weight in oxazalone-induced allergic dermatitis, a change that was accompanied by a reduction in inflammatory cells in the dermis and a decrease in tumor necrosis factor-alpha and interleukin-1alpha levels in the oxazalone-treated epidermis. These studies demonstrate that topically applied peroxisome proliferator activated receptor-alpha agonists possess receptor mediated, anti-inflammatory activity in both irritant and allergic contact dermatitis animal models. The anti inflammatory properties of peroxisome proliferator activated receptor-alpha agonists, coupled with their anti-proliferative and pro-differentiating effects, suggest that they could be beneficial for the treatment of a variety of cutaneous diseases. PMID- 11851882 TI - Quasi-normal cornified cell envelopes in loricrin knockout mice imply the existence of a loricrin backup system. AB - The cornified cell envelope, a lipoprotein layer that assembles at the surface of terminally differentiated keratinocytes, is a resilient structure on account of covalent crosslinking of its constituent proteins, principally loricrin, which accounts for up to 60%-80% of total protein. Despite the importance of the cell envelope as a protective barrier, knocking out the loricrin gene in mice results in only mild syndromes. We have investigated the epidermis and forestomach epithelium of these mice by electron microscopy. In both tissues, corneocytes have normal-looking cell envelopes, despite the absence of loricrin, which was confirmed by immunolabeling, and the absence of the distinctive loricrin containing keratohyalin granules (L-granules). Isolated cell envelopes were normal in thickness (approximately 15 nm) and mass per unit area (approximately 7.3 kDa per nm2); however, metal shadowing revealed an altered substructure on their cytoplasmic surface. Their amino acid compositions indicate altered protein compositions. Analysis of these data implies that the epidermal cell envelopes have elevated levels of the small proline-rich proteins, and cell envelopes of both kinds contain other protein(s) that, like loricrin, are rich in glycine and serine. These observations imply that, in the absence of loricrin, the mechanisms that govern cell envelope assembly function normally but employ different building-blocks. PMID- 11851883 TI - Epidermal stem cells do not communicate through gap junctions. AB - Although enrichment of putative epidermal stem cells has been achieved, a need for additional markers that can enable isolation of live keratinocytes is crucial for characterization of these cells. Earlier work has shown that connexin proteins are absent from basal cells in the limbal epithelium, a region of the corneal epithelium enriched in corneal stem cells. Accordingly, we investigated whether connexin 43, a gap junction protein present in the basal layer of normal human epidermis, can serve as a negative marker for keratinocyte stem cells. In humans, cells with immunohistochemically undetectable levels of connexin 43 are found in the epidermal basal layer of neonatal foreskin and in the follicular bulge region. About 10% of the basal keratinocytes are connexin 43 negative, as determined by flow cytometry. These cells are uniformly small and low in granularity. Restricted gap junction communication was confirmed by the failure of low molecular weight dyes to transfer between cells. Experiments carried out in mouse epidermis demonstrated that most of the slowly cycling cells, detected as label-retaining cells, do not express connexin 43. Thus, presumptive keratinocyte stem cells can be identified and separated based on connexin 43 expression. PMID- 11851884 TI - A close-up view of migrating Langerhans cells in the skin. AB - Langerhans cells of the epidermis and dermal dendritic cells screen the skin for invading antigens. They initiate primary immune responses after migrating from sites of antigen uptake to lymphoid organs. The skin is a feasible model to study the morphology and regulation of dendritic cell migration. We therefore used murine skin explant cultures for tracking the pathways of dendritic cell migration by electron microscopy. Several novel observations are reported. (i) In 48 h cultures of epidermal sheets numerous Langerhans cells migrated out between keratinocytes extending long and thin cytoplasmic processes ("veils"). (ii) Langerhans cells in transition from epidermis to dermis were observed by transmission electron microscopy. Where Langerhans cells penetrated the basement membrane, the lamina densa was focally absent. (iii) This was highlighted by scanning electron microscopy, which presented the basement membrane as a tightly packed and dense network of fibrils. (iv) Scanning electron microscopy of the dermis revealed dendritic cells extending their cytoplasmic processes and clinging to collagen fibrils. (v) Entry of dendritic cells into dermal lymphatics was observed by transmission electron microscopy. It occurred by transmigration through intercellular spaces of adjacent endothelial cells. Entry through wide gaps between endothelial cells also seemed to take place. (vi) Dendritic cells inside the afferent lymphatics frequently carried material such as melanosomes and apoptotic bodies. These observations visualize the cumbersome pathway that dendritic cells have to take when they generate immunity. PMID- 11851885 TI - Tyrosinase gene expression is regulated by p53. AB - Tyrosinase, the rate-limiting enzyme for melanin synthesis, is induced after ultraviolet irradiation as part of the tanning response, the major recognized photoprotective response of human skin. Other DNA-damaging agents and DNA fragments such as thymidine dinucleotides also induce tyrosinase gene expression. Moreover, like ultraviolet light they also activate p53. To determine whether p53 activation is required for this increased tyrosinase expression, we employed two experimental systems: (i) a human melanoma line (WM35) known to express wild-type p53 versus WM35 cells engineered to express a transcriptionally inactive dominant negative p53 (WM35-p53DN) or the empty vector alone (WM35-pCMV7) and (ii) mice with wild-type p53 versus p53 knockout mice. In WM35-p53DN cells, the baseline p53 protein level was higher than in WM35 or WM35-pCMV7 cells, and tyrosinase transcripts were lower. After ultraviolet irradiation, in all cell lines the p53 protein level increased within the first 24 h, as expected; and at 24 h tyrosinase mRNA levels were decreased. Consistent with the literature, these data in combination suggest that increased p53 protein level downregulates tyrosinase mRNA. In WM35 and WM35-pCMV7 cells at 48 and 72 h, however, whereas p53 levels remained elevated, tyrosinase mRNA levels compared to pre-irradiation levels tripled, whereas in WM35-p53DN cells levels remained below baseline. In thymidine dinucleotide-treated WM35 and WM35-pCMV7 cells there was a comparable upregulation of tyrosinase mRNA within 24 h that persisted through 72 h, but there was no upregulation of tyrosinase mRNA in WM35-p53DN cells any time after ultraviolet irradiation or thymidine dinucleotide treatment. In ear skin of p53 wild-type mice, topical application of thymidine dinucleotide induced a 4-5-fold increase in epidermal melanin content after 3 wk, but in p53 knockout mice thymidine dinucleotide application caused no detectable increase in melanin. Together, these data demonstrate that p53 activation increases tyrosinase mRNA level and subsequently pigmentation. The data further suggest that tanning is part of a p53-mediated adaptive response of mammalian skin to DNA damage from ultraviolet irradiation. PMID- 11851886 TI - Retinoic acid inhibits downregulation of DeltaNp63alpha expression during terminal differentiation of human primary keratinocytes. AB - Recently, the p53 homolog p63 has been implicated in sustaining the epidermal stem cell population. The p63 gene encodes six major products with transactivating or dominant-negative properties. The expression pattern of these isoforms in keratinocytes was investigated here. Northern blot, ribonuclease protection assay, reverse transcription-polymerase chain reaction, and western blot techniques sensitive for all six p63 isotypes verified the predominant expression of the truncated and potentially dominant-negative isotype DeltaNp63alpha in human keratinocytes. The expression of this isoform is downregulated when proliferating human primary keratinocytes begin to differentiate after growth factor withdrawal. The onset of differentiation does not change the ratio of two other weakly expressed isotypes DeltaNp63gamma and TAp63alpha relative to DeltaNp63alpha. Treatment of primary human keratinocytes with all-trans retinoic acid does not alter the expression pattern of p63 isotypes but prevents its downregulation as observed in control cell cultures. These data suggest that p63 expression in human keratinocytes is affected by all trans retinoic acid and this influence might contribute to the fine tuned keratinocyte proliferation and differentiation equilibrium in the mammalian epidermis. PMID- 11851887 TI - Hepoxilin B3 and its enzymatically formed derivative trioxilin B3 are incorporated into phospholipids in psoriatic lesions. AB - In previous studies we observed that normal human epidermis forms 12-oxo eicosatetraenoic acid (12-oxo-ETE) and hepoxilin B3 (HxB3) as major eicosanoids, both being elevated in psoriasis. We also observed that normal epidermis, in a reaction probably catalyzed by 12-lipoxygenase, only synthesize one of the two possible 10-hydroxy epimers of HxB3. We have now extended these previous studies investigating further transformation of HxB3 into trioxilin B3 (TrXB3) and esterification of both into phospholipids. Phospholipids were extracted from normal epidermis and from psoriatic scales. A combination of high performance liquid chromatography and gas chromatography-mass spectrometry analysis demonstrated the occurrence of HxB3 and TrXB3 in the phospholipids of psoriatic lesions. Alkaline- and phospholipase-A2-mediated hydrolysis of the phospholipids yielded similar quantities of both HxB3 and TrXB3 indicating their preference for the sn-2 position of glycerophospholipids. The thin layer chromatography analysis of the phospholipid classes after incubation of epidermal cells with [14C] labeled HxB3, TrXB3, 12-hydroxy-eicosatetraenoic acid (12-HETE), 12-oxo-ETE, or 15-HETE showed that 12-HETE was the most esterified (12-HETE >15-HETE > TrXB3 > 12-oxo-ETE > HxB3). HxB3 and TrXB3 were mainly esterified in phosphatidyl-choline and phosphatidyl-ethanolamine. HxB3 was also enzymatically converted into TrXB3 in vitro. HxB3 epoxide hydrolase-like activity was not observed when boiled tissue was incubated with [14C]-HxB3, this activity being located in the cytosol fraction (100,000 x g supernatant) of fresh tissue. These findings suggest that in vivo some part of HxB3 is transformed into TrXB3 and both compounds are partially incorporated into the phospholipids. PMID- 11851888 TI - Overexpression of serpin squamous cell carcinoma antigens in psoriatic skin. AB - Squamous cell carcinoma antigen belongs to the serpin family and is used for the diagnosis and management of squamous cell carcinoma. We investigated the involvement of squamous cell carcinoma antigen in psoriasis, as it is always detected in the sera of patients with psoriasis. Squamous cell carcinoma antigen localization in psoriatic epidermis varied depending on its concentration in the patient's sera. When its level was low in serum, weak and scattered staining was observed in the granular layer. With a high concentration of squamous cell carcinoma antigen, strong staining through the suprabasal to granular layer and condensed staining around the plasma membrane or intracellular space was detected in the affected epidermis. Interestingly, squamous cell carcinoma antigen was abundant in nuclei of the granular layer cells and elongated rete ridges. Immunoelectron microscopy confirmed the localization of squamous cell carcinoma antigen in the nuclei as well as in the periphery of the cell membrane. A cDNA library was constructed from psoriatic epidermis and both clones, SCCA1 and SCCA2, were obtained. Attempts to raise specific antibodies or to prepare cRNA probes for SCCA1 and SCCA2 were unsuccessful because of their nearly identical structures. A primer pair from each reactive site sequence enabled us to give a distinctive product for SCCA1 and SCCA2 by reverse transcription polymerase chain reaction. Analysis using these primers demonstrated that the SCCA2 transcript was specifically expressed in psoriatic skin tissues. Our results suggest that overexpression of squamous cell carcinoma antigens is associated with the disease activity of psoriasis. PMID- 11851889 TI - Promoter polymorphisms of the genes encoding tumor necrosis factor-alpha and interleukin-1beta are associated with different subtypes of psoriasis characterized by early and late disease onset. AB - The psoriatic inflammatory process is characterized by an overexpression of pro inflammatory cytokines such as tumor necrosis factor-alpha and interleukin-1beta compared with a relative deficiency of anti-inflammatory factors such as interleukin-10 and the interleukin-1 receptor antagonist (interleukin-1Ra). Gene polymorphisms that affect cytokine production may contribute to the disease associated cytokine imbalance and influence susceptibility to psoriasis. Here, we investigated the relationship between polymorphisms in the genes encoding for tumor necrosis factor-alpha (G-238A, G-308A), interleukin-1beta (C-511T, T+3953C), and interleukin-1Ra (intron 2), and cytokine production in peripheral blood mononuclear cells of healthy donors, and analyzed the distribution of these polymorphisms in patients with psoriasis vulgaris (n = 231) and healthy controls (n = 345). Carriage of tumor necrosis factor A-238 allele 2 (-238*A) was associated with increased production of tumor necrosis factor-alpha in response to lipopolysaccharide in vitro, and with early onset disease (< 40 y), especially in male patients with psoriasis [32% vs 7% in male controls; odds ratio = 6.78, 95% confidence interval = (3.18-15.15), p(adjusted) = 2 x 10(-7)]. Carriage of the interleukin-1B-511*1 (-511*C) homozygous genotype was associated with increased production of interleukin-1Ra in response to lipopolysaccharide and interleukin-10, and with late onset psoriasis [> or = 40 y; 61% vs 44% in controls; odds ratio = 2.04, 95% confidence interval = (1.19-3.53), p(adjusted) = 0.0419]. These findings indicate that gene polymorphisms associated with altered cytokine responses in vitro may modify age of onset of psoriasis. They also provide further evidence that patients with early and late onset psoriasis differ in their genetic background. PMID- 11851890 TI - Metabolic fate and selective utilization of major fatty acids in human sebaceous gland. AB - The sebaceous gland is an integral part of the pilosebaceous unit of mammalian skin, which produces and secretes a unique mixture of lipids, known as sebum. Wax esters, which account for approximately 25% of human sebaceous lipids, are unique in that they are not synthesized by other cells in the body. To explore the biosynthesis of wax esters, the metabolic fate of exogenously supplied saturated (16:0, 18:0), mono-unsaturated Delta9 (16:1, 18:1), and polyunsaturated (18:2, Delta9,12) fatty acids was followed in biopsy punches from human facial skin rich in sebaceous glands. Acetate was incorporated into all of the cellular and secreted lipids and 16:0 was incorporated into all of the fatty-acid-containing lipids. The 16:0 was elongated to 18:0 and the 16:1 was incorporated primarily into polar lipids, secondarily into triglycerides, but not into other lipids and was elongated to 18:1 (Delta11). As proven by HPTLC analysis, both 18:0 and 18:1 were incorporated into the cellular lipids but at a lower rate into wax esters. Moreover, addition of exogenous 18:1 was not further processed following initial incorporation. Linoleic acid (18:2, Delta9,12) was the only fatty acid tested that appeared to be subjected to beta-oxidation. This was proven to be specific to linoleic acid, as it did not induce the oxidation of other fatty acids. The ability of the sebaceous cells to synthesize wax esters correlated with the beta oxidation activity in these cells. Thus, the oxidation of linoleic acid is specific for the sebaceous cells and correlates with their function and differentiation. Our results provide evidence that the sebaceous gland selectively utilizes fatty acids as 16:0 is the preferred fatty acid that is incorporated into wax esters and linoleic acid undergoes beta-oxidation. PMID- 11851891 TI - Activation of nickel-specific CD4+ T lymphocytes in the absence of professional antigen-presenting cells. AB - Allergic contact dermatitis ensues from exaggerated T cell responses to haptens. Dendritic cells are required for the initiation of hapten sensitization, but they may not be necessary for disease expression. Here we investigated the antigen presenting cell requirement of nickel-specific CD4+ lymphocytes isolated from the blood of six allergic individuals. A significant proportion (42 out of 121; 35%) of the T cell clones proliferated in vitro to nickel also in the absence of professional antigen-presenting cells, suggesting a direct T-T hapten presentation. Antigen-presenting-cell-independent T cells showed a predominant T helper 1 phenotype. Nickel recognition by these T cells was major histocompatibility complex class II restricted, not influenced by CD28 triggering, independent from their state of activation, and did not require processing. The capacity of this T cell subset to be directly stimulated by nickel was not due to unique antigen-presenting properties, as both antigen presenting-cell-dependent and antigen-presenting-cell-independent clones displayed comparable levels of HLA-DR, CD80, and CD86, and were equally capable of presenting nickel to antigen-presenting-cell-independent clones. In contrast, neither T cell types activated antigen-presenting-cell-dependent T lymphocytes. T T presentation induced T cell receptor downregulation, CD25, CD80, CD86, and HLA DR upregulation, and interferon-gamma release, although to a lesser extent compared to those induced by dendritic cell-T presentation. Following T-T presentation, the clones did not undergo unresponsiveness and maintained the capacity to respond to dendritic cells pulsed with antigen. In aggregate, our data suggest that antigen-presenting-cell-independent T cell activation can effectively amplify hapten- specific immune responses. PMID- 11851892 TI - Ultraviolet photoproduct levels in melanocytic nevi and surrounding epidermis in human skin in situ. AB - Melanocytic nevi are localized benign proliferations of melanocytes. The number of nevi has been shown to be the major risk marker for the development of cutaneous melanoma. This study compares the induction of photoproducts in nevi and in surrounding skin after exposure to solar-simulating radiation. Cyclobutane pyrimidine dimers (TT=T and TT=C) and 6-4 photoproducts (TT-T and TT-C) were measured in 20 nevi and 20 surrounding skin samples obtained from 14 subjects, using a 32P-postlabeling method. The amount of all four types of photoproducts in nevi was found to be 3-5-fold lower than that in surrounding skin, and the difference was statistically significant (paired t test, p < 0.01). In nevi, the photoproduct level was significantly associated with the color of nevi (the lowest level in the darkest color of nevi; r = -0.86, p < 0.01 for TT=T; r = 0.68, p < 0.01 for TT=C). Our findings suggest that the magnitude of the DNA damage is not a sole risk marker for the development of cutaneous melanoma. PMID- 11851893 TI - Deletion of the cytoplasmatic domain of BP180/collagen XVII causes a phenotype with predominant features of epidermolysis bullosa simplex. AB - BP180/collagen XVII is a hemidesmosomal transmembrane molecule serving as cell surface receptor. Mutations in its gene cause junctional epidermolysis bullosa. Here, we report a patient with mutations in the gene for BP180/collagen XVII, COL17A1, but predominant phenotypic features of epidermolysis bullosa simplex. At birth, the proband presented with bullous lesions on the trunk, face, and hands. Ultrastructurally, hemidesmosomes were fairly normal, but the attachment of intermediate filaments with the hemidesmosomal plaques appeared to be impaired. Blister formation demonstrated both intraepidermal and junctional cleavage. Immunofluorescence staining with antibodies to keratins, several hemidesmosomal proteins, and the extracellular domain of BP180/collagen XVII showed normal staining patterns, whereas an antibody against the intracellular domain of BP180/collagen XVII yielded a negative immunofluorescence signal. Analysis of BP180/collagen XVII cDNA revealed a 1172 bp deletion corresponding to an in-frame deletion from Ile-18 to Asn-407 from the intracellular domain of the polypeptide. Mutation analysis of the COL17A1 gene disclosed a paternal nonsense mutation, R1226X, and a large maternal genomic deletion extending from intron 2 to intron 15, but no mutations in basal keratin genes. These findings underline the functional importance of the intracellular BP180/collagen XVII domain for the interaction of hemidesmosomes with keratin intermediate filaments and for the spatial stability of basal keratinocytes, and provide a functional explanation for the epidermolysis-bullosa- simplex-like phenotype. Further, the data demonstrate that defects in a given gene can cause unexpected phenotypes of epidermolysis bullosa categories, depending on the function of the affected protein domain. PMID- 11851894 TI - Increased mortality rate and not impaired ribosomal biogenesis is responsible for proliferative defect in dyskeratosis congenita cell lines. AB - X-linked dyskeratosis congenita is a rare inherited disorder mainly characterized by progressive changes in proliferating epidermal, mucosal, and bone marrow tissues that commonly emerge after 10 y of life. It is caused by mutations of the DKC1 gene, which codes for dyskerin, a protein that may play a role in ribosomal biogenesis. In order to verify whether the defects of proliferating tissues observed in dyskeratosis congenita are due to an altered ribosome synthesis, we studied ribosomal biogenesis in relation to cell proliferation in two lymphoblastoid cell lines from dyskeratosis congenita patients and in one control line. We observed that in the dyskeratosis congenita cell lines the rRNA transcription and maturation and proliferative capability remained unimpaired. Increasing the number of cell cycles, however, leads to a steep rise in the apoptotic fraction of dyskeratosis congenita cells, which is not observed in controls. These findings demonstrate that whereas dyskeratosis congenita cell lines do not display proliferation defects, they do show progressively increasing levels of apoptosis in relation to the number of cell divisions. This concept is consistent with (i) the delayed onset of dyskeratosis congenita proliferating tissue defects, which do not emerge during embrional development as would be expected with ribosomal biogenesis alterations, and (ii) with the increasing severity of the proliferating-tissue defects over time. PMID- 11851895 TI - Immunologic and histopathologic characterization of an active disease mouse model for pemphigus vulgaris. AB - Pemphigus vulgaris is an autoimmune blistering disease of the skin and mucous membranes that is caused by anti-desmoglein 3 IgG autoantibodies. Recently, we generated an active disease mouse model for pemphigus vulgaris by adoptive transfer of splenocytes from immunized desmoglein 3-/- mice to Rag2-/- mice. In this study, we performed immunologic and histopathologic studies using this pemphigus vulgaris model in mice and compared the gross and microscopic phenotypes of pemphigus vulgaris model mice and desmoglein 3-/- mice. Pemphigus vulgaris model mice showed strong in vivo IgG, and weak IgA deposition on keratinocyte cell surfaces in stratified squamous epithelia, and produced circulating anti-desmoglein 3 IgG antibodies without apparent cross-reactivity to desmoglein 1, in enzyme-linked immunosorbent assays. The predominant IgG subclass was IgG1. Pemphigus vulgaris model mice and desmoglein 3-/- mice were almost indistinguishable in terms of both gross and microscopic findings. Both types of mice showed suprabasilar acantholysis in the stratified squamous epithelia, including the oral mucous membranes and traumatized skin around the snout or paws; however, some pemphigus vulgaris model mice demonstrated a more severe phenotype than desmoglein 3-/- mice. The esophagus and forestomach were affected in some pemphigus vulgaris model mice, but not in desmoglein 3-/- mice. Furthermore, eosinophilic spongiosis, which is found in early pemphigus vulgaris lesions in patients, was observed in pemphigus vulgaris model mice but not in desmoglein 3-/- mice. Pemphigus vulgaris model mice reflect several of the histopathologic and immunologic features seen in pemphigus vulgaris patients, and provide a valuable tool to investigate the pathophysiologic mechanisms of pemphigus vulgaris. PMID- 11851896 TI - Depigmentation of brown Guinea pig skin by topical application of methimazole. PMID- 11851897 TI - Hepatitis C and human immune deficiency coinfection at the era of highly active antiretroviral therapy. AB - Interactions between human immunodeficiency virus (HIV) and hepatitis C virus (HCV) have been widely studied before the introduction of highly active antiretroviral therapies (HAART). We reviewed the potential impact of HAART on hepatitis C as well as the interactions between HIV and HCV therapies. Physicians should be aware of the potential risk of: (i) symptomatic liver disease in HCV HIV-coinfected patients at the era of triple antiretroviral therapy; (ii) potential liver deterioration paralleling immune restoration; (iii) lack of impact of triple antiretroviral therapy on HCV load; and (iv) potential drug related hepatitis which may modify the natural history of HCV-related liver disease. Liver biopsies should be performed regularly in these patients in order to identify patients with severe liver disease who require early initiation of anti-HCV therapy under close monitoring of their immune status. Treatment is, to date, based on the combination of ribavirin and interferon with an expected sustained response rate around 25%. An important unresolved issue is to better delineate the temporal place of anti-HCV and anti-HIV antiviral therapies. At least in coinfected patients with significant liver disease, namely necro inflammatory activity and/or fibrosis >or= 2, we believe that anti-HCV therapy is the priority since it lessens the risk of drug-induced hepatitis and of hepatitis due to immune restoration. PMID- 11851898 TI - Expression of the hepatitis C virus structural proteins in mammalian cells induces morphology similar to that in natural infection. AB - Like many positive-strand RNA viruses, replication of the hepatitis C virus (HCV) is associated with cytoplasmic membrane rearrangements. However, it is unclear which HCV proteins induce these ultrastructural features. This work examined the morphological changes induced by expression of the HCV structural proteins, core, E1 and E2, expressed from a Semliki Forest Virus (SFV) recombinant RNA replicon. Electron microscopy of cells expressing these proteins showed cytoplasmic vacuoles containing membranous and electron-dense material that were distinct from the type I cytoplasmic vacuoles induced during SFV replicon replication. Immunogold labelling showed that the core and E2 proteins localized to the external and internal membranes of these vacuoles, but at times were also associated with some of the internal amorphous material. Dual immunogold labelling with antibodies raised against the core protein and against an endoplasmic reticulum (ER)-resident protein (protein disulphide isomerase) showed that the HCV-induced vacuoles were associated with ER-labelled membranes. This report has identified an association between the HCV core and E2 proteins with induced cytoplasmic vacuoles which are morphologically similar to those observed in HCV-infected liver tissue, suggesting that the HCV structural proteins may be responsible for the induction of these vacuoles during HCV replication in vivo. PMID- 11851899 TI - Quantitative and functional differences in CD8+ lymphocyte responses in resolved acute and chronic hepatitis C virus infection. AB - CD8+ T lymphocyte responses are important in the clearance of viral infections. In chronic infections they may contribute to pathogenesis. To investigate the role of CD8+ T lymphocyte responses in viral clearance and chronic hepatitis C we have compared hepatitis C virus (HCV) specific cytotoxicity and interferon-gamma (IFN-gamma) production in patients with resolved-acute, and chronic HCV infection. CD8+ T cell responses to a panel of 13 HCV T cell peptide epitopes were studied using Elispot assays of IFN-gamma production and chromium release cytotoxicity assays. Responses of seven patients with resolved acute HCV infection were compared with those of 14 chronically infected patients. HCV specific cytotoxicity differentiated the two populations of patients. The majority (71%) of patients with resolved acute infection tested positive to 42% of relevant peptides compared with the minority (28%) of patients with chronic hepatitis C (P=0.03) who responded to only 8% of relevant peptides (P=0.0009). In contrast, HCV-specific IFN-gamma production was detected in 86% of patients with either resolved or chronic infection in response to 42% and 35%, respectively, of relevant peptides tested (not significant). In patients with chronic infection the magnitude of the HCV-specific IFN-gamma production was inversely correlated to viral load (R2=0.52; P=0.042). Failure to clear HCV infection may be attributable to the presence of noncytolytic IFN-gamma producing CD8+ T lymphocytes in chronically infected patients. However these CD8+ T cells may play a beneficial role in contributing to the control of viral load in chronic hepatitis C. PMID- 11851900 TI - Hypervariable region 1 quasispecies in hepatitis C virus genotypes 1b and 3 infected patients with normal and abnormal alanine aminotransferase levels. AB - The role of hepatitis C virus (HCV) heterogeneity in the severity of chronic hepatitis C infection remains unclear. Our aim was to study the hypervariable region 1 (HVR1) heterogeneity in patients with chronic hepatitis C infected with genotype 1b or 3 and with normal or abnormal alanine aminotransferase (ALT). HVR1 quasispecies were assessed by single strand conformational polymorphism (SSCP) in 67 patients with chronic hepatitis C, including 35 with persistently normal ALT and 32 with abnormal ALT. Sixty-two patients underwent a liver biopsy. Among the 67 patients, 40 were infected with genotype 1b and 27 with genotype 3. In univariate analysis, low heterogeneity ( 1:2048) were observed for more than 5 years following seroconversion in 80% (8/10) of seroconverters with sufficient information, indicating the development of persistent infection in these subjects. Three (37.5%) of the eight sero converters with persistent infection had fairly consistent, albeit mild, alanine aminotransferase elevations (30-130 IU/L) during the study. Anti-HCV seroconversions occurred at a very high rate in this community-based population in Japan, in which this infection is endemic. Persistence also developed at a high frequency among the cases of newly acquired infection, although the associated liver enzyme abnormalities were mild. PMID- 11851903 TI - World-wide epidemiology of HBeAg-negative chronic hepatitis B and associated precore and core promoter variants. AB - Hepatitis B is a serious disease that is endemic in many parts of the world. A significant proportion of patients with chronic hepatitis B (CHB) are infected with a variant form of the hepatitis B virus (HBV) which decreases or abolishes the production of hepatitis B e-antigen (HBeAg). The purpose of this literature review is to describe the epidemiology of HBeAg-negative CHB (e-CHB) worldwide. A literature search was conducted to identify studies pertaining to e-CHB and underlying variants (precore and core promoter). Fifty studies were included in our analysis. The median prevalence of e-CHB among patients with chronic HBV infection was 33% in the Mediterranean, 15% in Asia Pacific, and 14% in the USA and Northern Europe. The pre core stop codon variant was detected in a median of 60% (range 0-100%) of HBeAg-negative patients overall, 92% in the Mediterranean, 50% in Asia Pacific and 24% in the USA and Northern Europe. There were very few data on the prevalence of core promoter variants outside Asia where the median prevalence among HBeAg-negative patients was 77%. This literature review revealed that e-CHB is more common than previously suspected and that it is present worldwide with marked variations in the prevalence of associated HBV variants across different geographical regions. Additional research using population based samples of adequate size based on a consensus definition of e-CHB and using standardized HBV DNA assays is needed to better estimate the true prevalence of e CHB and its associated HBV variants. PMID- 11851904 TI - Epidemiological changes in hepatitis C virus genotypes in France: evidence in intravenous drug users. AB - Hepatitis C virus (HCV) genotypes are distributed differently depending on geography and route of infection. We characterized the distribution of genotypes in a large cohort of patients with chronic hepatitis C in the South-east of France and evaluated the relative prevalence according to time of acquisition. One thousand, one hundred-and-eighty-three patients who were anti-HCV-positive were studied. HCV genotype distribution has changed significantly from the 1960s to 2000. The prevalence of genotype 1b decreased from 47% before 1978 to 18.8% in the 1990s while the prevalence of genotype 1a and 3a increased during the same period from 18% and 15.3% to 28.8% and 26.3%, respectively. The logistic regression model showed that genotype 1a was significantly more common in patients infected through intravenous drug injection odds ratio ((OR): 2.08, P < 0.01) and after 1990 (OR: 1.98, P < 0.05). Genotype 1b was significantly less frequent in patients infected through intravenous drug injection (OR: 0.17, P < 0.001) and has decreased since 1978 (OR: 0.27, P < 0.001). Genotype 3a was independently associated with intravenous drug injection (OR: 6.1, P < 0.001) and tattooing (OR: 8.01, P < 0.001) and was more frequent in the 1979-90 period (OR: 2.05 and 1.74, P < 0.001 and P < 0.05). Our results show a modification of HCV genotypes distribution over the last four decades due to an increase of intravenous drug use (IVDU) contamination and an evolution of HCV genotypes distribution only in IVDU population characterized by a decrease of genotype 1b, an increase of genotype 3a from 1970 to 1990 and a higher increase of genotype 1a which is currently the predominant genotype in our population. PMID- 11851905 TI - Progressive hepatic fibrosis in healthy carriers of hepatitis C virus with a transaminase breakthrough. AB - In short-term studies, patients with chronic hepatitis C virus (HCV) infection, consistently normal serum aminotransferase (ALT) levels, and minimal or mild necro-inflammatory changes in the liver, did not progress to histologically severe hepatitis. There are no data on longer term outcome of liver disease in these patients. We describe two patients with HCV infection (genotype 2c) with a rise in serum ALT values greater than 10 times the upper normal value that occurred after an 8- and 15-year period of persistently normal or minimally elevated ALT levels. In both patients, the rise in ALT values lasted more than 16 weeks and was not associated with any symptom or risk factor for acute hepatitis. A liver biopsy performed 4 and 18 months after the ALT flare showed clear-cut progression from chronic hepatitis with mild activity to chronic hepatitis with severe activity and central to portal septal fibrosis (Ishak score: grading 14 and 6; staging: 4 and 5, respectively). Hence, extended surveillance of HCV carriers with consistently normal or minimally elevated ALT values is warranted as these patients are at risk of ALT flares and may develop progressive liver disease. PMID- 11851906 TI - Sarcoidosis in two patients with chronic hepatitis C treated with interferon, ribavirin and amantadine. AB - We report two patients with chronic hepatitis C, both nonresponders to a previous course of interferon (IFN), who developed or suffered an exacerbation of sarcoidosis while under treatment with IFN-alpha2a, ribavirin and amantadine. Patient 1: symptoms appeared after week 4 and treatment was withdrawn at month 9 due to severe weight loss, marked dyspnea, muscular weakness, dryness of mouth and facial paralysis. Stage III pulmonary sarcoidosis and polyneuropathy were confirmed. The patient had become steroid dependent and nine months after cessation of the treatment dyspnea and muscular weakness still persisted. She achieved a complete sustained response of hepatitis C. Patient 2: presented with a previous diagnosis of granulomatous hepatitis with chronic active hepatitis C and chronic dermatitis. The treatment exacerbated a cutaneous sarcoidosis. Furthermore, hiliar adenopathies consistent with stage I sarcoidosis became evident. Sarcoidosis responded to corticosteroids, but elevated transaminases and hepatitis C viraemia resisted. Hence, the combination of amantadine with ribavirin and IFN can develop or exacerbate subclinical sarcoidosis. A synergistic effect of these three drugs is suggested. PMID- 11851907 TI - The role of auxin-binding protein 1 in the expansion of tobacco leaf cells. AB - Tobacco leaf was used to investigate the mechanism of action of auxin-binding protein 1 (ABP1). The distributions of free auxin, ABP1, percentage of leaf nuclei in G2 and the amount of auxin-inducible growth were each determined in control tobacco leaves and leaves over-expressing Arabidopsis ABP1. These parameters were compared with growth of tobacco leaves, measured both spatially and temporally throughout the entire expansion phase. Within a defined window of leaf development, juvenile leaf cells that inducibly expressed Arabidopsis ABP1 prematurely advanced nuclei to the G2 phase. The ABP1-induced increase in cell expansion occured before the advance to the G2 phase, indicating that the ABP1 induced G2 phase advance is an indirect effect of cell expansion. The level of ABP1 was highest at the position of maximum cell expansion, maximum auxin inducible growth and where the free auxin level was the lowest. In contrast, the position of maximum cell division correlated with higher auxin levels and lower ABP1 levels. Consistent with the correlations observed in leaves, tobacco cells (BY-2) in culture displayed two dose-dependent responses to auxin. At a low auxin concentration, cells expanded, while at a relatively higher concentration, cells divided and incorporated [3H]-thymidine. Antisense suppression of ABP1 in these cells dramatically reduced cell expansion with negligible effect on cell division. Taken together, the data suggest that ABP1 acts at a relatively low level of auxin to mediate cell expansion, whereas high auxin levels stimulate cell division via an unidentified receptor. PMID- 11851908 TI - Functional importance of conserved domains in the flowering-time gene CONSTANS demonstrated by analysis of mutant alleles and transgenic plants. AB - CONSTANS promotes flowering of Arabidopsis in response to long-day conditions. We show that CONSTANS is a member of an Arabidopsis gene family that comprises 16 other members. The CO-Like proteins encoded by these genes contain two segments of homology: a zinc finger containing region near their amino terminus and a CCT (CO, CO-Like, TOC1) domain near their carboxy terminus. Analysis of seven classical co mutant alleles demonstrated that the mutations all occur within either the zinc finger region or the CCT domain, confirming that the two regions of homology are important for CO function. The zinc fingers are most similar to those of B-boxes, which act as protein-protein interaction domains in several transcription factors described in animals. Segments of CO protein containing the CCT domain localize GFP to the nucleus, but one mutation that affects the CCT domain delays flowering without affecting the nuclear localization function, suggesting that this domain has additional functions. All eight co alleles, including one recovered by pollen irradiation in which DNA encoding both B-boxes is deleted, are shown to be semidominant. This dominance appears to be largely due to a reduction in CO dosage in the heterozygous plants. However, some alleles may also actively delay flowering, because overexpression from the CaMV 35S promoter of the co-3 allele, that has a mutation in the second B-box, delayed flowering of wild-type plants. The significance of these observations for the role of CO in the control of flowering time is discussed. PMID- 11851909 TI - Two glucosyltransferases are involved in detoxification of benzoxazinoids in maize. AB - Benzoxazinoids are major compounds involved in chemical defence in grasses. These toxins are stored in the vacuole as glucosides. Two glucosyltransferases, BX8 and BX9, that catalyse this last step of benzoxazinoid biosynthesis have been isolated via functional cloning. No close relative of these maize genes was found among the known glucosyltransferases. The enzymes display a very high degree of substrate specificity. DIMBOA, the major benzoxazinoid in young maize, is the preferred substrate. Both genes are highly expressed in young maize seedlings, the developmental stage with the highest activity of benzoxazinoid biosynthesis. Bx8 is included in the cluster of DIMBOA biosynthesis genes located on the short arm of chromosome 4. Hence, the gene cluster comprises three different enzymatic functions and a complete set of genes for the biosynthesis of DIBOA glucoside. Bx9 mapped to chromosome 1. Expression of Bx8 and Bx9 in Arabidopsis corroborated the potency of the enzymes in detoxification of their substrates. This capacity might have implications for allelopathic interactions. PMID- 11851910 TI - Characterization of plant proliferating cell nuclear antigen (PCNA) and flap endonuclease-1 (FEN-1), and their distribution in mitotic and meiotic cell cycles. AB - The biochemical and cell cycle-dependent properties of proliferating cell nuclear antigen (OsPCNA) and flap endonuclease-1 (OsFEN-1) were characterized from rice (Oryza sativa). OsPCNA was physically associated with OsFEN-1 and increased the flap-endonuclease activity of OsFEN-1 by 2.5-fold. Northern and Western blotting analysis revealed that OsPCNA and OsFEN-1 were present in meristematic tissues such as cultured cells, shoot apical meristem and root apical meristem. No expression was detected in the mature leaves, although they were exposed to UV. Both of these proteins were localized in the nuclei of the interphase cells including G1, S and G2, and in the nuclear region at telophase. The distribution patterns of plant PCNA and FEN-1 in meiotic cell progression were investigated using microsporocytes of lily (Lilium longiflorum cv. Hinomoto). During the leptotene to pachytene stages, PCNA and FEN-1 were localized in the nuclear region. The florescence gradually disappeared from diplotene to metaphase I. Interestingly, signals for PCNA formed 10-20 intense spots at leptotene. The number of spots decreased to 1-5 at zygotene and finally to 1 at pachytene. The roles of OsPCNA and OsFEN-1 in mitotic and meiotic cell cycles are discussed. PMID- 11851911 TI - ABA plays a central role in mediating the regulatory effects of nitrate on root branching in Arabidopsis. AB - The formation of lateral roots (LR) is a major post-embryonic developmental event in plants. In Arabidopsis thaliana, LR development is inhibited by high concentrations of NO3(-). Here we present strong evidence that ABA plays an important role in mediating the effects of NO3(-) on LR formation. Firstly, the inhibitory effect of NO3(-) is significantly reduced in three ABA insensitive mutants, abi4-1, abi4-2 and abi5-1, but not in abi1-1, abi2-1 and abi3-1. Secondly, inhibition by NO3(-) is significantly reduced, but not completely abolished, in four ABA synthesis mutants, aba1-1, aba2-3, aba2-4 and aba3-2. These results indicate that there are two regulatory pathways mediating the inhibitory effects of NO3(-) in A. thaliana roots. One pathway is ABA-dependent and involves ABI4 and ABI5, whereas the second pathway is ABA-independent. In addition, ABA also plays a role in mediating the stimulation of LR elongation by local NO3(-) applications. PMID- 11851912 TI - Infra-red thermography revealed a role for mitochondria in pre-symptomatic cooling during harpin-induced hypersensitive response. AB - The establishment of Erwinia amylovora harpin-induced hypersensitive response (HR) in Nicotiana sylvestris was followed by infra-red thermography (IRT). Three to four hours after elicitation, the temperature decreased in the harpin infiltrated zone associated to stomatal opening. The marked drop in temperature which reached 2 degrees C and preceded necrosis symptoms for several hours, is thus likely caused by higher transpiration. Neither of these effects was observed in a respiratory mutant, affected in complex I structure and function and over expressing alternative oxidase, indicating that they are directly or indirectly mediated by mitochondrial function. However, as the HR establishment was similar in both wild type and mutant, cell death was either uncorrelated with the observed epidermal changes or occurred by a different signalling pathway in the two genotypes. IRT revealed a novel aspect of plant-pathogen interactions and could be applied to screen for mutants affected in elicitor signalling and/or for respiratory mutants. PMID- 11851913 TI - Gene targeting in Arabidopsis. AB - Precise modification by gene targeting (GT) provides an important tool for studies of gene function in vivo. Although routine with many organisms, only isolated examples of GT events have been reported for flowering plants. These were at low frequencies precluding reliable estimation of targeting efficiency and evaluation of GT mechanisms. Here we present an unambiguous and straightforward system for detection of GT events in Arabidopsis using an endogenous nuclear gene encoding protoporphyrinogen oxidase (PPO), involved in chlorophyll and heme syntheses. Inhibition of PPO by the herbicide Butafenacil results in rapid plant death. However, the combination of two particular mutations renders PPO highly resistant to Butafenacil. We exploited this feature for selection of GT events by introducing the mutations into the PPO gene by homologous recombination. We have estimated the basal GT frequency to be 2.4 x 10(-3). Approximately one-third of events were true GT (TGT) leading to the anticipated modification of the chromosomal PPO copy. The remaining events could be classified as ectopic GT (EGT) arising by modification of vector DNA by the chromosomal template and its random integration into the Arabidopsis genome. Thus the TGT frequency in our experimental setup is 0.72 x 10(-3). In view of the high efficiency of Arabidopsis transformation, GT experiments of a reasonable size followed by a PCR screen for GT events should also allow for modification of non selectable targets. Moreover, the system presented here should contribute significantly to future improvement of GT technology in plants. PMID- 11851914 TI - Hydroxyl radical-induced cell-wall loosening in vitro and in vivo: implications for the control of elongation growth. AB - Hydroxyl radicals (OH) are capable of unspecifically cleaving cell-wall polysaccharides in a site-specific reaction. I investigated the hypothesis that cell-wall loosening underlying the elongation growth of plant organs is controlled by apoplastically produced OH attacking load-bearing cell-wall matrix polymers. Isolated cell walls (operationally, frozen/thawed, abraded segments from coleoptiles or hypocotyls, respectively) from maize, cucumber, soybean, sunflower or Scots pine seedlings were pre-loaded with catalytic Cu or Fe ions and then incubated in a mixture of ascorbate + H2O2 for generating OH in the walls. This treatment induced irreversible wall extension (creep) in walls stretched in an extensiometer. The reaction could be promoted by acid pH and inhibited by several OH scavengers. Generation of OH by the same reaction in living coleoptile or hypocotyl segments caused elongation growth. Auxin-induced elongation growth of maize coleoptiles could be inhibited by OH scavengers. Auxin promoted the production of superoxide radicals (O2(-)), an OH precursor, in the growth-controlling outer epidermis of maize coleoptiles. It is concluded that OH fulfils basic criteria for a wall-loosening factor acting in auxin-mediated elongation growth of plant species with widely differing cell-wall polysaccharide compositions. PMID- 11851915 TI - Chromosome painting in Arabidopsis thaliana. AB - Chromosome painting, that is visualisation of chromosome segments or whole chromosomes based on fluorescence in situ hybridization (FISH) with chromosome specific DNA probes is widely used for chromosome studies in mammals, birds, reptiles and insects. Attempts to establish chromosome painting in euploid plants have failed so far. Here, we report on chromosome painting in Arabidopsis thaliana (n = 5, 125 Mb C(-1)). Pools of contiguous 113-139 BAC clones spanning 2.6 and 13.3 Mb of the short and the long arm of chromosome 4 (17.5 Mb) were used to paint this entire chromosome during mitotic and meiotic divisions as well as in interphase nuclei. The possibility of identifying any particular chromosome region on pachytene chromosomes and within interphase nuclei using selected BACs is demonstrated by differential labelling. This approach allows us, for the first time, to paint an entire autosome of an euploid plant to study chromosome rearrangements, homologue association, interphase chromosome territories, as well as to identify homeologous chromosomes of related species. PMID- 11851916 TI - Divalent cations and polyamines bind to loop 8 of 14-3-3 proteins, modulating their interaction with phosphorylated nitrate reductase. AB - Binding of 14-3-3 proteins to nitrate reductase phosphorylated on Ser543 (phospho NR) inhibits activity and is responsible for the inactivation of nitrate reduction that occurs in darkened leaves. The 14-3-3-dependent inactivation of phospho-NR is known to require millimolar concentrations of a divalent cation such as Mg2+ at pH 7.5. We now report that micromolar concentrations of the polyamines, spermidine(4+) and spermine(3+), can substitute for divalent cations in modulating 14-3-3 action. Effectiveness of the polyamines decreased with a decrease of polycation charge: spermine(4+) > spermidine(3+) >>> cadavarine(2+) approximately putrescine(2+) approximately agmatine(2+) approximately N1 acetylspermidine(2+), indicating that two primary and at least one secondary amine group were required. C-terminal truncations of GF14 omega, which encodes the Arabidopsis 14-3-3 isoform omega, indicated that loop 8 (residues 208-219) is the likely cation-binding site. Directed mutagenesis of loop 8, which contains the EF hand-like region identified in earlier studies, was performed to test the role of specific amino acid residues in cation binding. The E208A mutant resulted in a largely divalent cation-independent inhibition of phospho-NR activity, whereas the D219A mutant was fully Mg(2+)-dependent but had decreased affinity for the cation. Mutations and C-terminal truncations that affected the Mg(2+) dependence of phospho-NR inactivation had similar effects on polyamine dependence. The results implicate loop 8 as the site of divalent cation and polyamine binding, and suggest that activation of 14-3-3s occurs, at least in part, by neutralization of negative charges associated with acidic residues in the loop. We propose that binding of polyamines to 14-3-3s could be involved in their regulation of plant growth and development. PMID- 11851917 TI - PkMADS1 is a novel MADS box gene regulating adventitious shoot induction and vegetative shoot development in Paulownia kawakamii. AB - Direct regeneration of shoot buds in vitro is an important technique in plant genetic manipulation. We describe the isolation and functional characterization of a novel MADS box cDNA (PkMADS1) from Paulownia kawakamii leaf explants undergoing adventitious shoot regeneration. mRNA gel blot analysis confirmed the expression of PkMADS1 in the shoot-forming cultures, but no signal was observed in the callus-forming cultures. PkMADS1 transcripts were also detected in shoot apices, but not in root apices, initial leaf explants or the flower. In situ hybridization revealed that its expression was restricted to developing shoot primordia in the excised leaf cultures, suggesting a role for this gene in adventitious shoot formation. Transgenic Paulownia plants over-expressing the PkMADS1 gene showed some changes in phenotype, such as axillary shoot formation. In the antisense transformants, shoots were stunted and had altered phyllotaxy, and, in some lines, the shoot apical meristem appeared to have been used up early during shoot development. Leaf explants from the antisense transgenic plants showed a tenfold decrease in shoot regeneration compared with explants from sense transformants or wild-type. Our results show that PkMADS1 is a regulator of shoot morphogenesis. PMID- 11851918 TI - Pistillody, homeotic transformation of stamens into pistil-like structures, caused by nuclear-cytoplasm interaction in wheat. AB - Homeotic transformation of stamens into pistil-like structures (pistillody) has been observed in a cytoplasmic substitution (alloplasmic) line of wheat (Triticum aestivum L.) cv. Norin 26, which has the cytoplasm of a wild relative species, Aegilops crassa L. On the other hand, an alloplasmic line of wheat cv. Chinese Spring (CS) with Ae. crassa cytoplasm has normal flowers. This is due to the presence in the CS nucleus of a fertility-restoring gene, Rfd1. Deletion mapping analysis revealed that Rfd1 is located on the middle part of the long arm of chromosome 7B. To investigate the function of the Rfd1 gene by a loss-of-function strategy, we produced alloplasmic lines of CS ditelosomic 7BS [(cr)-CSdt7BS] and CS monotelodisomic 7BS [(cr)-CSmd7BS] with the Ae. crassa cytoplasm, and characterized their phenotypes. The line (cr)-CSdt7BS without Rfd1 exhibited pistillody in all florets, and also female sterility. Scanning electron microscopy of the young spikes revealed that the pistillody was induced at an early stage of stamen development. The pistillate stamens often developed incomplete ovule-like structures with integuments instead of tapetum and pollen grains. It is possible that MADS box genes are associated with the induction of pistillody, because the expression of wheat APETALA3 homologue (WAP3) was reduced in the young spikes of (cr)-CSdt7BS. In addition, a histological study indicated that the female sterility in (cr)-CSdt7BS is due to the abnormality of the ovule, which fails to form an inner epidermis and integuments in the chalaza region. The line (cr)-CSmd7BS, hemizygous for Rfd1, showed partial pistillody (51%) and restored female fertility up to 72%. These results suggest that the induction of both pistillody and ovule deficiency caused by the Ae. crassa cytoplasm is inhibited by the Rfd1 gene in a dose-dependent manner. PMID- 11851919 TI - FLC, a repressor of flowering, is regulated by genes in different inductive pathways. AB - The MADS-box protein encoded by FLOWERING LOCUS C (FLC) is a repressor of flowering. Loci in the autonomous flowering pathway control FLC levels. We show the epistatic groupings of autonomous pathway mutants fca/fy and fve/fpa, based on their effects on flowering time, are consistent with their effects on FLC transcript and protein levels. We demonstrate that synergistic increases in FLC mRNA and protein expression occur in response to interactions between the autonomous pathway mutants fca and fpa and mutants in other pathways (fe, ft, fha) that do not regulate FLC when present as single mutants. These changes in FLC levels provide the molecular basis of the interactions previously shown in genetic analyses. The interactions between genes of multiple pathways emphasize the central position of FLC in the control of floral initiation. FLC protein levels match those of its mRNA for a range of genetic, developmental and environmental variables, indicating that control of FLC is at the level of transcription or transcript stability. The autonomous and photoperiod pathways also interact at the level of SOC1. FLC acts as a repressor of SOC1, and SOC1 levels are low when FLC levels are high. In C24 plants which have moderately high FLC levels, flowering occurs without a decrease in FLC level, but the SOC1 level does increase. Thus SOC1 levels can be upregulated through the activities of other pathways, despite the repression by FLC. PMID- 11851920 TI - RNA splicing in higher plant mitochondria: determination of functional elements in group II intron from a chimeric cox II gene in electroporated wheat mitochondria. AB - Higher plant mitochondria mainly contain group II introns presenting a secondary structure with six helical domains linked to a central hub. Experimental evidence of functional elements in higher plant mitochondria introns is limited since they are unable to undergo self-splicing and the definition of functional domains is based on data obtained from yeast autocatalytic introns. Here we study the role of putative functional elements required for the splicing reaction. The exon binding and intron-binding sites (EBS and IBS, respectively), and the domain 6, which is involved in lariat formation, were analysed by site-directed mutagenesis and transient expression in electroporated mitochondria. The data presented here demonstrate the role of EBS1-IBS1 and EBS2-IBS2 interactions and reveal a new secondary-structure interaction. The role of the C to U editing conversion in the IBS1 motif is discussed. PMID- 11851921 TI - Recognition specificity of self-incompatibility maintained after the divergence of Brassica oleracea and Brassica rapa. AB - The determinants of recognition specificity of self-incompatibility in Brassica are SRK in the stigma and SP11/SCR in the pollen, respectively. In the pair of S haplotypes BrS46 (S46 in B. rapa) and BoS7 (S7 in B. oleracea), which have highly similar SRK alleles, the SP11 alleles were found to be similar, with 96.1% identity in the deduced amino acid sequence. Two other pairs of S haplotypes, BrS47 and BoS12, and BrS8 and BoS32, having highly similar SRK and SP11 alleles between the two species were also found. The haplotypes in each pair are considered to have been derived from a single S haplotype in the ancestral species. The allotetraploid produced by interspecific hybridization between homozygotes of BrS46 and BoS15 showed incompatibility with a BoS7 homozygote and compatibility with other B. oleracea S haplotypes in reciprocal crossings. This result indicates that BrS46 and BoS7 have maintained the same recognition specificity after the divergence of the two species and that amino acid substitutions found in such cases in both SRK alleles and SP11 alleles do not alter the recognition specificity. DNA blot analysis of SRK, SP11, SLG and other S-locus genes showed different DNA fragment sizes between the interspecific pairs of S haplotypes. A much lower level of sequence similarity was observed outside the genes of SRK and SP11 between BrS46 and BoS7. These results suggest that the DNA sequences of the regions intervening between the S-locus genes were diversified after or at the time of speciation. This is the first report demonstrating the presence of common S haplotypes in different plant species and presenting definite evidence of the trans-specific evolution of self incompatibility genes. PMID- 11851922 TI - Trehalose-6-phosphate synthase 1, which catalyses the first step in trehalose synthesis, is essential for Arabidopsis embryo maturation. AB - Despite the recent discovery that trehalose synthesis is widespread in higher plants very little is known about its physiological significance. Here we report on an Arabidopsis mutant (tps1), disrupted in a gene encoding the first enzyme of trehalose biosynthesis (trehalose-6-phosphate synthase). The tps1 mutant is a recessive embryo lethal. Embryo morphogenesis is normal but development is retarded and stalls early in the phase of cell expansion and storage reserve accumulation. TPS1 is transiently up-regulated at this same developmental stage and is required for the full expression of seed maturation marker genes (2S2 and OLEOSN2). Sucrose levels also increase rapidly in seeds during the onset of cell expansion. In Saccharomyces cerevisiae trehalose-6-phosphate (T-6-P) is required to regulate sugar influx into glycolysis via the inhibition of hexokinase and a deficiency in TPS1 prevents growth on sugars (Thevelein and Hohmann, 1995). The growth of Arabidopsis tps1-1 embryos can be partially rescued in vitro by reducing the sucrose level. However, T-6-P is not an inhibitor of AtHXK1 or AtHXK2. Nor does reducing hexokinase activity rescue tps1-1 embryo growth. Our data establish for the first time that an enzyme of trehalose metabolism is essential in plants and is implicated in the regulation of sugar metabolism/embryo development via a different mechanism to that reported in S. cerevisiae. PMID- 11851926 TI - The evolution and function of central venous catheters for dialysis. AB - The use of central venous catheters (CVC) for acute or chronic dialysis has been a relatively recent innovation in Nephrology. The first problem addressed has been how to allow removal and return of blood at high flow rates throughout a dialysis treatment. Four solutions have emerged: place the lumens within the right atrium; place the removal lumen on the inside of the catheter; use a large catheter size; or provide independent limbs with multiple blood-entry ports to draw and return blood in all directions. Many other requirements include resistance to infection, especially the passage of organisms around the catheter. A subcutaneous Dacron cuff within a tunnel has successfully accomplished this goal for most chronic CVC dialysis catheters, but other immobilizing devices such as plugs have also been successful. Materials for CVC dialysis catheters have improved, providing strength to allow the catheters to last for several years, with flexibility to avoid vein damage (in general). However, component and material failures still occur, and some materials are incompatible with medications placed at the exit site. CVC for dialysis will remain a necessary choice for many patients beginning and continuing dialysis therapy. PMID- 11851925 TI - Catheter access for hemodialysis: an overview. AB - The provision of hemodialysis requires repeated, reliable access to the central circulatory system. Long-term hemodialysis has best been provided by arteriovenous fistulae and arteriovenous grafts. In recent years, more and more patients have been chronically dialyzed with tunneled dialysis catheters. These catheters, which were originally developed as a short-term bridge to permanent vascular access, have made up an increasing percentage of maintenance vascular access. While these catheters have the advantage of ease of placement and are immediately ready for use, they substantially increase the risk of bacteremia, stenosis of central veins, and even mortality. PMID- 11851927 TI - Catheter performance. AB - Venous catheters differ from peripheral arteriovenous (AV) access devices in many important ways. This discussion focuses on their performance as a conduit for blood flow between the patient and the dialyzer and on how catheter function is both limited and enhanced relative to the more common peripheral accesses. Catheter flow is limited by the high resistance inherent in the extended length of venous catheters relative to dialysis needles, but the high rate of flow in central veins also diminishes the opportunity for access recirculation. Cardiopulmonary recirculation is absent in patients with catheter access unless the patient also has a peripheral access. In the latter case, the same detrimental effect on urea clearance is seen regardless of which access device is used. Flow-dependent recirculation through circuits other than the peripheral AV access reduces the efficiency of dialysis (regardless of the type of access, catheter, or peripheral AV device used) across both catheters and peripheral AV devices. The inside diameter of the catheter plays a sensitive role in determining catheter resistance to flow. Slight increases in diameter under the same pressure head are associated with large increases in flow. Negative pressure at the catheter inflow port generated by the blood pump is magnified relative to peripheral devices, predisposing to partial collapse of the pump tubing segment and erroneous blood flow readings by the pump motor speed indicator. Setting a limit on prepump negative pressure can minimize this error. Future applications of dialysis may require lower pump speeds, which would allow more liberal use of catheter access if their potential for infection and clotting can be reduced. PMID- 11851928 TI - Acute dialysis catheters. AB - Acute dialysis catheters are non-cuffed, non-tunnelled catheters used for immediate vascular access. They are primarily used for acute renal failure in bed bound patients and for short-term use in patients with malfunction of permanent access. Long-term use of acute catheters is not recommended, but does occur with acceptable infection rates in dialysis centers where tunnelled, cuffed catheters are not available. Most acute catheters are made of polyurethane, but silicone catheters are now available with larger lumen sizes capable of delivering blood flow rates over 300 ml/min. Acute catheters should be inserted in the internal jugular or femoral vein under ultrasound guidance to minimize complications. Subclavian catheters cause stenosis, thrombosis, and perforation of vessels. Intermittent catheter malfunction still occurs, particularly for left-sided internal jugular catheters and catheters placed in women. Blood flow may improve with lumen reversal, intraluminal t-PA, or guidewire exchange. Limiting duration of use and dressing catheter exit sites with dry gauze and antiseptic ointments can prevent catheter-related infections. PMID- 11851929 TI - Chronic catheter placement. AB - Since the 1997 publication of the Disease Outcomes Quality Initiative (DOQI) vascular access guidelines for cuffed, tunneled catheter placement, additional evidence supporting these recommendations has been published, including additional documentation supporting the right internal jugular vein as the preferred site for insertion. Placing the catheter tip in the right atrium rather than in the superior vena cava will provide adequate blood flow to support effective hemodialysis. The right atrial positioning of the catheter tip will also accommodate catheter tip retraction and decrease the likelihood of malfunction. Overwhelming evidence now supports the use of ultrasound guidance to assist cannulation of the internal jugular vein. This evidence is based on several studies documenting anatomical variations of the internal jugular vein. Ultrasound guidance has significantly decreased the incidence of serious complications of jugular vein cannulation. Finally, a specific technique of catheter placement with variations for catheter types is described. PMID- 11851930 TI - Catheter thrombosis. AB - Catheter malfunction due to poor flow is a common problem. When it occurs early, the cause is generally technical. Late occurrences are most often related to thrombus formation. Several types of thrombus may be seen, differing by location and supposed mechanism of formation. The most common offender, however, is the fibrin sheath thrombus. Prevention of catheter malfunction is an endeavor that continues to beg many questions. Catheter malfunction should be treated early to avoid inadequate dialysis. In the past, urokinase was a highly valuable aid to the nephrologist in managing this problem. Since this agent became unavailable a suitable alternative has not emerged. Recombinant tissue plasminogen activator (tPA) seems to be the most likely candidate for this role; however, it is not currently available in a packaging form that is optimal for this purpose. Currently, catheter exchange appears to be the best available alternative for this problem, which cannot yet be resolved by simpler means. PMID- 11851931 TI - Central venous dialysis catheters: catheter-associated infection. AB - Tunneled dialysis catheters (TDC) are extensively used for long-term venous hemodialysis access and their use is frequently associated with infectious complications. Catheter-related bacteremia (CRB) is the most common and important infection associated with TDC use and may be caused by a wide variety of Gram positive or Gram-negative organisms. Prevention of CRB can be difficult despite use of rigorous infection-control techniques for catheter insertion and access. A number of antibacterial catheter-packing solutions hold promise for reduction of CRB. Treatment of CRB with antibiotics alone yields poor results and may increase the risk for other infectious complications, especially endocarditis. In selected cases where initial infection control can be achieved with antibiotics, guidewire exchange of the TDC results in cure rates equivalent to those of TDC removal and subsequent replacement. Dialysis programs should monitor TDC infections with attention to incidence, bacteriology, and outcomes. PMID- 11851932 TI - Subcutaneous vascular access devices. AB - Tunneled dialysis catheters have played an important role in providing vascular access for hemodialysis for over 20 years, yet limitations associated with poor flow, thrombosis, and infections due to the transcutaneous nature of catheters have led to the development of alternative vascular access devices. This article reviews two historical devices-the Hemasite and the Bentley DiaTAP button-and provides an overview of two new fully subcutaneous vascular access devices-the LifeSite Hemodialysis Access System (Vasca, Inc., Tewksbury, MA) and the Dialock Hemodialysis System (Biolink Corp., Norwell, MA). These new subcutaneous devices differ significantly from currently available tunneled dialysis catheters in their design and method of use and may result in improved outcomes in hemodialysis patients. Indeed, initial clinical experiences with this new class of fully implanted access devices validate a subcutaneous approach to hemodialysis access and suggest that their high flow rates and low complication rates may make subcutaneous devices an attractive alternative to dialysis catheters for hemodialysis patients. PMID- 11851934 TI - Access blood flow: debate continues. PMID- 11851935 TI - Access blood flow: debate continues. PMID- 11851938 TI - Platelet chemokines and their receptors: what is their relevance to platelet storage and transfusion practice? AB - The role of platelets as inflammatory cells is demonstrated by the fact that they can release many growth factors and inflammatory mediators, including chemokines, when they are activated. The best known platelet chemokine family members are platelet factor 4 (PF4) and beta-thromboglobulin (beta-TG), which are synthesized in megakaryocytes, stored as preformed proteins in alpha-granules and released from activated platelets. However, platelets also contain many other chemokines such as interleukin-8 (IL-8), growth-regulating oncogene-alpha(GRO-alpha), epithelial neutrophil-activating protein 78 (ENA-78), regulated on activation normal T expressed and secreted (RANTES), macrophage inflammatory protein-1alpha (MIP-1alpha), and monocyte chemotactic protein-3 (MCP-3). They also express chemokine receptors such as CCR4, CXCR4, CCR1 and CCR3. Platelet activation is a feature of many inflammatory diseases such as heparin-induced thrombocytopenia, acquired immunodeficiency syndrome, and congestive heart failure. Substantial amounts of PF4, beta-TG and RANTES are released from platelets on activation, which may occur during storage. Although very few data are available on the in vivo effects of transfused chemokines, it has been suggested that the high incidence of adverse reactions often observed after platelet transfusions may be attributed to the chemokines present in the plasma of stored platelet concentrates. PMID- 11851939 TI - HTLV antibody screening using mini-pools. AB - At the present time, the UK blood transfusion services do not screen blood donations for anti-HTLV. This presentation describes a pilot study to ascertain the feasibility of HTLV antibody screening using mini-pools and also provides an estimate of HTLV prevalence within our donor population in Scotland and Northern Ireland. The Abbott/Murex HTLV I/II GE80/81 ELISA was selected for the trial. Thirty confirmed HTLV positive library samples were tested at various dilutions and five were shown to be nonreactive at a dilution of 1:100. Residues of mini pools (of up to 95 individual donations) prepared for HCV NAT testing were tested with the Abbott/Murex GE80/81 assay. Of 6666 mini-pools (equivalent to 570 609 donations) tested, six were repeatedly reactive. All six mini-pools were confirmed HTLV antibody positive by line immunoassay. Four were confirmed to be HTLV-I positive, one HTLV-II positive and one HTLV positive (unable to type). Dilutions (1:100) of the five HTLV "nonreactive" positive samples were included in each test plate and used to determine a grey-zone cut-off. Using this grey zone system an additional six (0.09%) mini-pool samples gave repeatedly reactive grey-zone results, none of which were confirmed. The minimum Scottish/Irish HTLV donor prevalence was shown to be 1:95 000. PMID- 11851940 TI - Transfusion practice in Helsinki University Central Hospital: an analysis of diagnosis-related groups (DRG). AB - Transfusion data combined with data automatically recorded in hospital databases provides an outstanding tool for blood utilization reporting. When the reporting is performed with an online analytical processing (OLAP) tool, real time reporting can be provided to blood subscribers. When this data is combined with a common patient classification system, Diagnosis-Related Groups (DRG), it is possible to produce statistical results, that are similar in different institutions and may provide a means for international transfusion bench-marking and cost comparison. We use a DRG classification to describe the transfusion practice in Helsinki University Central Hospital. The key indicators include the percentage of transfused patients, the number of transfused units and costs in different DRG groups, as well as transfusion rates per DRG weighted treatment episodes. Ninety-three per cent of all transfusions could be classified into different DRGs. The largest blood-using DRG group was acute adult leukaemia (DRG 473), which accounted for 10.4% of all transfusion costs. The 13 largest blood consuming DRGs accounted for half the total costs in 1998. Currently, there is a lack of an internationally accepted standardized way to report institutional or national transfusion practices. DRG-based transfusion reporting might serve as a means for transfusion benchmarking and thus aid studies of variations in transfusion practice. PMID- 11851941 TI - Measurement of blood volume after haemodilution with haemoglobin-based oxygen carriers by a radiolabelled-albumin method. AB - Recent studies have shown that the use of haemoglobin-based oxygen-carrying solutions (HBOCs) for perioperative haemodilution could significantly reduce the need for packed red blood cells in clinical practice. Though the effects of HBOCs on plasma volume have been characterized in experimental models of volume resuscitation from hypovolaemic shock, little is known about their action in normovolaemic haemodilution conditions. We therefore applied a radiolabelled serumalbumin method to determine blood volume after haemodilution with crosslinked or conjugated haemoglobin, in comparison with a reference solution of hydroxyethyl starch (HES). Three groups of New Zealand white rabbits were studied (n = 7 each group) subjected to moderate exchange transfusion with low molecular weight HES, bis(3,5-dibromosalicyl)fumarate crosslinked haemoglobin (alphaalpha Hb), or dextran-conjugated haemoglobin (Hb-Dex-BTC). HES induced no changes in heart rate and blood pressure. The amplitude and duration of blood pressure increase and bradycardia were similar in both haemoglobin groups. A significant contraction of blood volume (12%) was observed 60 min after haemodilution with alphaalpha-Hb, compared to HES and Hb-Dex-BTC. At the same time point, a decrease in absolute haemoglobin (plasma haemoglobin x plasma volume) was also noted. This study suggests that in haemodilution conditions, the specific oncotic properties and circulating persistence of crosslinked and conjugated haemoglobin solutions affect the pattern of blood volume distribution differently. PMID- 11851942 TI - Identification and quantification of anti-D, -C and -G in alloimmunized pregnant women. AB - Our objectives were to investigate possible overestimation of maternal anti-D due to co-existing anti-C and/or anti-G, and to confirm the presence of anti-D in plasma presumed to contain anti-D+C. We investigated 96 samples (from 22 antenatal patients and 74 blood donors) initially identified as containing anti D+C using routine investigation procedures. Anti-D quantification was performed using an Astoria Pacific International 300 (API 300) continuous flow analyser with R1R1 and R2R2 reagent red cells. Where possible, samples were tested manually using a rare D+, C-, G- cell, to confirm the presence of anti-D. Fifty two of 96 samples (11/22 antenatal patients and 41/74 blood donors) gave >50% higher anti-D quantification results with R1R1 cells than with R2R2 cells. Anti-D was not detected using manual techniques in 16 of 73 samples tested (10/22 antenatal patients and 6/51 blood donors). Anti-D quantification using R1R1 reagent red cells may cause inaccurate estimation of anti-D levels, when anti-C and/or anti-G are present. Indeed, a significant number of cases, where apparent anti-D+C is identified, may contain only anti-C+G and lack an anti-D component. This may in turn lead to a failure to administer prophylactic anti-D immunoglobulin to RhD negative patients in cases where anti-D is not present, putting these patients at risk from immunization with possible consequences to future pregnancies. PMID- 11851943 TI - Epitope mapping of four novel CD44 monoclonal antibodies using surface plasmon resonance and soluble CD44. AB - CD44 is a ubiquitous multistructural and multifunctional cell surface adhesion molecule. The molecular diversity of this glycoprotein is generated by both post translational modification and the differential use of alternatively spliced exons which play a critical role in determining the exact conformation of the molecule. CD44 isoforms are found in many tissues and in soluble form in plasma. Soluble CD44 was purified by a two-step purification combining ion exchange and immuno-affinity chromatography. Our aim was to check that all the known antigenic epitopes are present on sCD44, which could thus be used for the mapping of new anti-CD44 antibodies. Competitive binding assays using reference antibodies and novel anti-CD44 monoclonal antibodies were performed by real-time biospecific interaction analysis. Reference mAbs identified on soluble CD44 the three distinct epitopes previously defined using red blood cell membrane CD44. From the four novel CD44 mAbs, two mAbs (NaM198-6B5, NaM198-10B4) mapped to epitope group 1, whereas the others (NaM10-8F4, NaM77-9D6) mapped to epitope group 2. Immunopurified sCD44 obtained from the plasma of healthy donors appears to be a usable tool for the mapping of anti-CD44 mAbs. PMID- 11851944 TI - Massive haemolysis in a group A recipient of a group O peripheral blood stem cell allogeneic transplant. AB - A 28-year-old man with lymphoblastic lymphoma received G-CSF mobilized stem cells from his HLA identical sister, who had been taking methotrexate for psoriasis until 1 month prior to harvest. The recipient's blood group was A Rh D positive and donor's group O Rh D positive. Engraftment and major haemolysis were evident by day 9. From day 9 to day 13 he received 17 units of blood (approximately 3 L of red cells) at a time when his calculated red cell volume was 1 L. This massive transfusion requirement was not explained by his clinical condition and led us to consider factors that may have influenced the degree of haemolysis. The stem cell graft contained 2.85 x 10(6) CD34+ cells kg(-1) and we speculate there was B cell hyperactivity following the withdrawal of methotrexate in the donor and this went unchecked by the omission of methotrexate in the GVHD prophylaxis of the recipient. We have also considered the phenomenon of bystander haemolysis, previously unreported in this situation, as haemolysis of transfused group O blood must have also occurred. The case also illustrates the importance of transfusing donor type red cells and recipient type fresh frozen plasma (FFP) and platelets into minor mismatched transplant patients. The decision to revert to donor type FFP and platelets should only be made when the direct antiglobulin test is negative and the appropriate isohaemagglutinins are no longer demonstrable. PMID- 11851945 TI - Autologous placental blood transfusion after a planned neonatal pacemaker implantation. AB - A fetal third-degree atrio-ventricular block was diagnosed in a 22-year-old woman at 24 weeks of gestation. During clinical follow-up a mild to moderate tricuspid regurgitation was detected at 35 weeks and maternal connective tissue diseases were excluded. Early postnatal cardiac pacing therapy was planned and autologous placental blood transfusion was proposed for the treatment of probable blood loss due to pacemaker implantation. A male infant was delivered at 38 weeks vaginally and 87 mL of placental blood was collected from the undelivered placenta. The placental blood was negative for viral markers and syphilis. Subsequent tests for bacteriological cultures were also negative. Within 6 h of delivery, the baby underwent cardiac pacemaker implantation and received 45 mL of autologous placental blood. Autologous placental blood transfusion was successfully used for the treatment of predicted blood loss after a planned neonatal surgical procedure. PMID- 11851946 TI - Early acute hepatitis B infection and hepatitis B vaccination in blood donors. PMID- 11851948 TI - Micronutrient supplementation for malaria control--hype or hope? PMID- 11851949 TI - Time travel with Oliver Twist--towards an explanation foa a paradoxically low mortality among recent immigrants. AB - First-generation immigrant populations in industrialized countries frequently have a lower mortality than the host population, a finding that is unexpected and often dismissed as the result of bias. We propose an alternative explanation for a real, albeit temporal, mortality advantage. We base our argument on two premises: First, that there are differences in the progression of the health transition between the immigrants' countries of origin and industrialized host countries; and, second, that there are differences in the speed at which changes in mortality from various causes occur after migration. Mortality from treatable communicable and maternal conditions, still high in many countries of origin, quickly declines to levels close to those of the host country. Mortality from ischaemic heart disease, the most common cause of death in the host countries, takes years or decades to rise to comparable heights. This is because of the time lag between increases in risk factor levels and an increased risk of coronary death. Hence, first-generation immigrants may initially experience a lower mortality than the host population, a point that has so far been under appreciated in discussions of immigrant mortality. After adopting a western lifestyle immigrants face an increasing risk of ischaemic heart disease. The increase occurs on top of a persisting risk from conditions associated with childhood deprivation, e.g. stomach cancer and stroke--the unfinished agenda of the health transition that immigrants experience. PMID- 11851950 TI - Factors associated with coverage in community-directed treatment with ivermectin for onchocerciasis control in Oyo State, Nigeria. AB - Community-directed distribution with ivermectin (CDTI) has been adopted by the African Programme for Onchocerciasis Control (APOC) as its main strategy for achieving sustained high coverage in endemic communities. This article describes the coverage results achieved when CDTI was introduced in four local government areas of Oyo State, Nigeria. Using a household survey after the second distribution, researchers documented that 68.6% of the community overall received the drug, and as did 85.0% of those who were eligible (not pregnant, not sick and at least 5 years of age). Six factors were associated with having received ivermectin. Four were personal characteristics: being male, being at least 35 years of age, belonging to the Fulani ethnic minority, having taken the drug at a previous distribution. Two village characteristics were smaller size, as measured by number of houses, and use of the central place mode of distribution as opposed to house-to-house. In-depth interviews with village leaders and volunteer community-directed distributors (CDDs) and focus group discussions among villagers provided qualitative data to help interpret the findings. Women in many villages felt excluded from decision making. The concerns of migrant farm workers living in Yoruba farm settlements were not well understood by health staff or the majority population. The main factor associated with receiving the ivermectin was having received it before, and qualitative comments about side-effects and beliefs about orthodox drugs indicated that issues of personal preferences, not addressed in a household coverage survey, need to be explored further. The findings can provide guidance in re-orienting health workers to the importance of fostering participation and cohesion among all segments of the community, especially the inclusion of women and minority groups. PMID- 11851951 TI - Malaria in pregnancy in rural Mozambique: the role of parity, submicroscopic and multiple Plasmodium falciparum infections. AB - BACKGROUND: Falciparum malaria affects pregnant women, especially primigravidae, but before malaria control programmes targeted to them can be designed, a description of the frequency and parity pattern of the infection is needed. There is little information on the frequency and effect of submicroscopic malaria infection, as well as on multiplicity of Plasmodium falciparum genotypes in pregnancy. This study aimed to describe the prevalence of malaria parasitaemia and anaemia and their relation to parity and age in pregnant women, during two malaria transmission seasons in a rural area of southern Mozambique. It also tried to assess the frequency and effect on anaemia of submicroscopic and multiple falciparum infections. METHODS: A total of 686 pregnant women were enrolled in three cross-sectional community-based surveys during different transmission seasons in rural southern Mozambique. In each survey a questionnaire was administered on previous parity history, the gestational age was assessed, the axillary temperature recorded and both haematocrit and malaria parasitaemia were determined. We used polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) analysis to determine submicroscopic and multiple P. falciparum infections in a subsample of women. FINDINGS: A total of 156 women (23%) had microscopic parasitaemia, of which 144 (92%) were asexual forms of P. falciparum. The prevalence of clinical malaria was 18 of 534 (3%), that of anaemia, 382 of 649 (59%). In a multivariate analysis age but not parity was associated with an increased risk of microscopic parasitaemia. Anaemia was associated with microscopic P. falciparum parasitaemia. Both malaria parasitaemia and anaemia were more frequent during the rainy season. Although not statistically significant, submicroscopic infections tended to be more frequent among grand-multiparous pregnant women. Subpatent infections were not associated with increased anaemia. Multiplicity of infection was not associated with either parity, age or anaemia. Likewise, there was no correlation between P. falciparum density and multiplicity of infection. INTERPRETATION: We did not observe a clear parity pattern of malaria and anaemia in our study. It is possible although unlikely that selection bias may have influenced these findings; but in which direction is unclear. The importance of locally based research before implementation of public health measures needs to be highlighted. According to our findings, a more cost-effective malaria control approach in this area would be targeting all pregnant women regardless of their parity. This would be also more feasible logistically as it would not rely on accurate ascertainment of parity, something that is not always easy in busy antenatal clinics. PMID- 11851952 TI - Weekly chloroquine prophylaxis and the effect on maternal haemoglobin status at delivery. AB - Our aim was to determine the effectiveness of chloroquine prophylaxis in reducing the frequency of malaria-induced anaemia at delivery. We estimated the haemoglobin levels of 207 parturients; 82 (39.6%) had been on chloroquine prophylaxis [treatment group (TG)] while 125 (60.4%) did not take any malaria preventive medication antenatally [control group (CG)]. The proportion of women with malaria parasitaemia was significantly higher in CG than TG [risk ratio (RR=1.57, CI=1.05-2.34)]. The dose-response relationship between the severity of parasitaemia and the risk of being anaemic (P < 0.001) confirms a strong correlation between gestational malaria and maternal anaemia. There was a 35% reduction in risk for anaemia in the TG compared with the CG (RR=0.65, 0.40 1.06). The difference in risk was more pronounced after adjusting for disparity in place of residence, educational status and obstetric history (adjusted RR=0.54, CI=0.21-0.98). Primiparous mothers appeared to have benefited more from the antianaemic effects of malaria chemoprevention than mothers of higher parity (protective effectiveness 43% compared with 33%, respectively). In conclusion, despite reports of widespread Plasmodium falciparum resistance to chloroquine on the African continent, malaria chemosuppression with the drug was found beneficial in reducing the risk of anaemia at delivery among Cameroonian women. PMID- 11851953 TI - Epidemiology and clinical manifestations of Leishmania donovani infection in two villages in an endemic area in eastern Sudan. AB - We conducted a longitudinal study in an endemic area for visceral leishmaniasis (VL) in eastern Sudan to compare the epidemiology and clinical spectrum of Leishmania donovani infection in two populations differing in ethnic background and duration of residence in the area. The study took place in two villages from April 1994 to April 1996. In Um-Salala village, which is inhabited by members of the Masaleet tribe, half of the villagers had previous exposure to cutaneous leishmaniasis (Leishmaria major) before moving there. The population of the second village, Mushrau Koka, belong to the Hausa tribe and most were born there. The incidence of VL was 20.4/1000 person-years in 1994/1995 and increased sharply to 38.3/1000 person-years in 1995/1996 in Um-Salala. A rise in the incidence of VL was also observed in Mushrau Koka but with a lower incidence, 3.3/1000 person years to 4.6/1000 person-years. The incidence rate of confirmed VL reflects only a limited part of the total infection rate which includes various forms of subclinical infection. The ratio of clinical to subclinical infection in Um Salala was 1.2 : 1 in 1994/1995 compared with 2.6 : 1 in 1995/1996. This ratio was 1 : 11 in 1994/1995 and 1 : 2.5 in 1995/1996 in Mushrau Koka. In both villages the mean age of subclinical cases was higher, but in Mushrau Koka the mean age of subclinical cases also was higher than that of subclinical cases in Um-Salala. The leishmanin skin test (LST) was positive in 56% of individuals in Um-Salala and in 33% in Mushrau Koka. VL only occurred in leishmanin-negative individuals. Post kala-azar dermal leishmaniasis (PKDL) followed in 58% of confirmed VL patients in Um-Salala; the low incidence of VL for Mushrau Koka did not permit to estimate a PKDL rate. The clinical manifestations resulting from exposure to L. donovani range from subclinical infection to VL and PKDL. No firm conclusion as to the difference in incidence of VL between the two villages could be reached but differences in exposure to VL and cutaneous leishmaniasis (CL) as well as other factors such as ethnic background and differences in nutritional status may play a role. PMID- 11851954 TI - Use of clinical algorithms for diagnosing malaria. AB - Several attempts have been made to identify symptoms and signs based algorithms for diagnosing malaria. In this paper, we review the results of published studies and assess the risks and benefits of this approach in different epidemiological settings. Although in areas with a low prevalence the risk of failure to treat malaria resulting from the use of algorithms was low, the reduction in the wastage of drugs was trivial. The odds of wastage of drugs increased by 1.49 (95% confidence limit 1.45-1.51) for each 10% decrease in the prevalence of malaria. In highly endemic areas the algorithms had a high risk of failure to treat malaria. The odds of failure to treat increased by 1.57 (95% confidence limit 1.50-1.65) for each 10% increase in the prevalence. Furthermore, the best clinical algorithms for diagnosing malaria were site-specific. We conclude that the accuracy of clinical algorithms for diagnosing malaria is not sufficient to determine whether antimalarial drugs should be given to children presenting with febrile illness. In highly endemic areas where laboratory support is not available, the policy of offering antimalarial drugs to all children presenting with a febrile illness recommended by the integrated child management initiative is appropriate. PMID- 11851955 TI - Detection of antigen B of Cysticercus cellulosae in cerebrospinal fluid for the diagnosis of human neurocysticercosis. AB - Neurocysticercosis (NCC) is a major cause of morbidity and mortality in developed and developing countries. The diagnosis of this disease remains a problem. We report the detection of specific antigenic fraction (antigen B) of Cysticercus cellulosae by enzyme-linked immunosorbent assay (ELISA) in various fractions of cerebrospinal fluid (CSF) obtained by high performance liquid chromatographic (HPLC) separation, for the diagnosis of human NCC. Forty patients attending or admitted to Nehru Hospital, Chandigarh were included in the study: 10 with suspected NCC, 20 with other neurological diseases and 10 undergoing surgery under spinal anaesthesia for non-neurological conditions, who served as controls. CSF samples collected from all patients and controls were subjected to chromatographic separation on an HPLC system. Antigen B (AgB) was detected in separated fractions by an ELISA test and compared with the detection of antibody response in CSF samples by indirect haemagglutination (IHA) technique. Antigen B was detected in 9 out of 10 patients with suspected NCC based on clinical symptoms and radioimaging reports, but in none of the control subjects. However, antigen B was also detected in 9 out of 20 patients with other neurological disorders, mostly tubercular meningitis. Antibody response by IHA was found positive in only 2 of 10 cases clinically suspected of NCC. In conclusion, antigen B detection in CSF samples may be a useful adjunct to clinical suspicion and radiological reports for the diagnosis of NCC as there is no gold standard criteria to confirm this disease. However, the test needs to be evaluated on more patients in countries where tuberculosis and cysticercosis are endemic due to the high cross reactivity with samples from tubercular meningitis patients. PMID- 11851956 TI - Resurgence in filarial transmission after withdrawal of mass drug administration and the relationship between antigenaemia and microfilaraemia--a longitudinal study. AB - Seven village units endemic for filariasis were assigned randomly into three arms with different intervention strategies in the years 1995 and 1996. Villages in Group A received two annual mass drug administrations (MDAs) of diethylcarbamazine (DEC) plus ivermectin (IVR). Group B received the same MDAs in combination with vector control; in Group C only placebo was administered. Post treatment evaluation in 1997 revealed marked reductions in microfilaraemia prevalences (MFP) and geometric mean intensities (GMI). Subsequently, vector control in Group B was continued by the community. Groups A and B received no MDAs between 1997 and 1999 and were re-evaluated in 1999. During this evaluation, antigenaemia prevalence (AGP) was estimated along with MFP, using immunochromatographic test (ICT) kits. The gains of the MDAs were sustained in Group B, while resurgences occurred in Group A, where annual transmission potential (ATP) rose from 21 to 631.6 and MFP doubled. Group C continued to have high ATPs (1057-1617), while Group B had very low ATPs (0-63). After Mulla's corrections, the reductions in MFP were 62.7 and 83.5% and for GMI 72.1 and 91.4% in Group A and B, respectively, compared with Group C. Vector control preserved the gains of MDAs and accounted for 55.8% and 67.1% reductions in MFP and GMI in Group B compared with Group A. The reductions in AGP were 29.4% in Group A and 39.2% in Group B against Group C. The differences between MFP and AGP were nearly uniform across all villages and there was a significant correlation (r=0.98) between the two variables. A similar significant relationship was observed between MFP and AGP values across age groups (r=0.95). Prediction of AGP values from MFP values was proposed with regression equations. We conclude that vector control would be useful as an adjuvant to chemotherapy to prevent resurgences. PMID- 11851957 TI - Evaluation of staff performance and material resources for integrated schistosomiasis control in northern Senegal. AB - BACKGROUND: A project to improve integrated control of schistosomiasis in the primary health care system of northern Senegal was implemented from February 1995 until September 1999, shortly after a Schistosoma mansoni outbreak. The activities included additional training of doctors and nurses in symptom-based treatment and making praziquantel (PZQ) available for an affordable price. OBJECTIVE: To investigate staff performance and the availability and costs of diagnostic materials and PZQ at the end of this intervention project. METHODS: We performed structured interviews with staff from 55 health care facilities in five districts. RESULTS: Respondents from 23 health care facilities reported both S. haematobium and S. mansoni in the coverage area, 32 reported only S. haematobium and three only S. mansoni. The average cost to patients for consultation, diagnosis, treatment and transportation to a referral health care facility was approximately 1.60 Euro. Fifty-seven per cent of the health care facilities with reported S. haematobium in the coverage area treated patients presenting with haematuria on symptoms; 56% of the health care facilities with reported S. mansoni in the coverage area treated patients presenting with blood in stool on symptoms. Thirteen per cent performed a diagnostic test for patients presenting with haematuria and 12% for patients presenting with blood in stool. The remainder, approximately one-third of the health care facilities, referred their patients to another facility for a diagnostic test. Implementation of symptom based treatment in all health care facilities will reduce the total costs by 0.43 Euro (29%) for patients infected with S. haematobium and 0.78 Euro (46%) for patients infected with S. mansoni. Of the 53 health care facilities with schistosomiasis in their area, 37 had PZQ in stock of which 33 (88%) sold PZQ for the recommended retail price of 0.15 Euro per tablet (or 0.60 Euro per course of four tablets) or lower. CONCLUSION: Four years after the start of the intervention project, patients presenting with schistosomiasis related symptoms can generally expect proper diagnosis and treatment at all levels of the health care system in Northern Senegal, either at the initial visited health care facility or after referral. However, a further reduction of the total costs of treatment is still possible by a better implementation of symptom-based treatment and further reduction of the costs of PZQ. PMID- 11851958 TI - The application of GIS technology to equitably distribute fieldworker workload in a large, rural South African health survey. AB - A method is presented that has numerous applications to health systems provision in developing countries where limited physical access to primary health care is a major factor contributing to the poor health of populations. An accessibility model within a geographical information system (GIS) is used to predict average inter-homestead walking times and subdivide the study area into units of equal completion time. The method could be used to ergonomically design home-based care and tuberculosis directly observed treatment programmes and inform the siting of health facilities. The paper highlights the use of GIS technology as a powerful tool in developing countries. PMID- 11851959 TI - Female genital cutting in southern urban and peri-urban Nigeria: self-reported validity, social determinants and secular decline. AB - Despite growing public resistance to the practice of female genital cutting (FGC), documentation of its prevalence, social correlates or trends in practice are extremely limited, and most available data are based on self-reporting. In three antenatal and three family planning clinics in South-west Nigeria we studied the prevalence, social determinants, and validity of self-reporting for FGC among 1709 women. Women were interviewed on social and demographic history, and whether or not they had undergone FGC. Interviews were followed by clinical examination to affirm the occurrence and extent of circumcision. In total, 45.9% had undergone some form of cutting. Based on WHO classifications by type, 32.6% had Type I cuts, 11.5% Type II, and 1.9% Type III or IV. Self-reported FGC status was valid in 79% of women; 14% were unsure of their status, and 7% reported their status incorrectly. Women are more likely to be unsure of their status if they were not cut, or come from social groups with a lower prevalence of cutting. Ethnicity was the most significant social predictor of FGC, followed by age, religious affiliation and education. Prevalence of FGC was highest among the Bini and Urhobo, among those with the least education, and particularly high among adherents to Pentecostal churches; this was independent of related social factors. There is evidence of a steady and steep secular decline in the prevalence of FGC in this region over the past 25 years, with age-specific prevalence rates of 75.4% among women aged 45-49 years, 48.6% among 30-34-year olds, and 14.5% among girls aged 15-19. Despite wide disparities in FGC prevalence across ethnic, religious and educational groups, the secular decline is evident among all social subgroups. PMID- 11851960 TI - 4D, or not 4D: that is the question. PMID- 11851961 TI - Prenatal testing: towards realistic expectations of patients, providers and policy makers. PMID- 11851962 TI - Receipt of information and women's attitudes towards ultrasound scanning during pregnancy. AB - OBJECTIVE: To audit women's receipt of information during routine antenatal ultrasound investigations; and to investigate women's perceptions of, and attitudes towards, routine screening. DESIGN: Analysis of the records of 384 women from in and around Nottingham, UK, who maintained a diary throughout the course of their pregnancies. RESULTS: The mean number of ultrasound scans received by each women during her pregnancy was 2.6, with more than 96% of women receiving at least one scan. Women initially scanned earlier received more scans in total. Women appeared aware of the specific reasons for each scan being undertaken, and only a minority were dissatisfied by the information they had received. The preponderance of information received during the scans related to results and arrangements for subsequent visits, with only 10% of women reporting receiving information about the procedure itself. Most women reported positive feelings towards scanning, and few gave indications that they would prefer changes in the conduct of the procedure. CONCLUSIONS: The great majority of the women in our study were satisfied with their experiences of routine ultrasound scanning, accepting the procedure uncritically. PMID- 11851963 TI - Maternal pain and anxiety in genetic amniocentesis: expectation versus reality. AB - OBJECTIVE: To investigate maternal perceptions of both pain and anxiety before and after genetic amniocentesis. STUDY DESIGN: This prospective study of midtrimester, singleton pregnancies was conducted between March 2000 and July 2000. Study variables included patient demographics, medical and obstetric histories, indication for amniocentesis and a description of the source of information used by the patient regarding the procedure and technical degree of difficulty. Maternal pain and anxiety associated with performing amniocentesis were subjectively quantified with the use of the visual analog scale (VAS). Statistical analysis included Wilcoxon signed rank test, anova, and simple and stepwise regression analyses. RESULTS: One hundred and eighty-three women participated in the study. Perception of pain before amniocentesis was significantly higher compared to that expressed immediately after the procedure, with a mean VAS score of 3.7 +/- 2.5 vs. 2.1 +/- 2.0 (P < 0.0001). Similarly, perception of anxiety was significantly greater prior to the procedure, with a mean VAS score of 4.6 +/- 2.8 vs. 2.8 +/- 2.4 after the amniocentesis (P < 0.0001). Perceptions of pain and anxiety were significantly and positively correlated to each other both before and after the procedure (P < 0.0001). History of a prior amniocentesis was the only variable associated with reducing expected pain and anxiety (negative correlation, P < 0.001), whereas the technical degree of difficulty was the only significant variable impacting on the actual pain and anxiety (positive correlation, P < 0.005). CONCLUSIONS: Preamniocentesis counseling should emphasize the fact that, for most women, the actual pain and anxiety experienced during the procedure are significantly lower than expected. In fact, on a scale of 0-10, the mean level of pain was only 2.1, with a slightly higher mean level of anxiety. PMID- 11851964 TI - Depressive reactions and stress related to prenatal medicine procedures. AB - OBJECTIVE: The experience of pregnancy is mainly influenced by the availability of prenatal screening procedures. Previous screening studies focus on somatic abnormalities. The psychological experience of the parents-to-be recedes into the background. The purpose of the present study was to analyze the emotional mood of pregnant women and their partners before and their psychological stress before and after different prenatal screening procedures (ultrasound, chorionic villus sampling or amniocentesis). METHODS: In the study, 140 pregnant women, and, in 108 cases, their partners, were asked to complete questionnaires before and immediately after the prenatal testing. Depending on the applied prenatal procedure, the subjects were assigned to: the invasive group (amniocentesis or chorionic villus sampling) and the noninvasive group (ultrasound group). We used the German version of the Centre for Epidemiological Studies-Depression Scale of Hautzinger and the short questionnaire of actual situative perceived stress of Muller. RESULTS: Before the prenatal examination, the mean level of depression of pregnant women was the highest in the noninvasive group compared to the invasive group, although the between-group difference was not significant. However, for their male partners, the mean level of depression was significantly different between the noninvasive and the invasive groups. Furthermore, women undergoing invasive diagnostic or noninvasive diagnostic procedures were significantly more depressed than their partners. The analysis of the actual stress ratings showed a significant reduction from the prescreening to the postscreening stress for pregnant women and their partners in both groups. CONCLUSIONS: This study reveals that the individual experience of prenatal diagnosis is not determined by the invasiveness of the procedure. Immediate visual presentation of the fetus and confirmation of a normal test result reduce stress that has previously been induced by the prenatal test itself. This contradiction should be discussed with the parents. PMID- 11851965 TI - Brains and faces in holoprosencephaly: pre- and postnatal description of 30 cases. AB - OBJECTIVE: To investigate the prenatal appearance of the holoprosencephaly spectrum. METHODS: A database of 1750 fetuses with congenital anomalies identified by ultrasound was prospectively collected from 1987 to 2000. Among them, 30 cases (1.7%) with holoprosencephaly were prenatally identified and described. RESULTS: The prevalence of holoprosencephaly in the Health Region of the National Center for Fetal Medicine in Norway was 1.26 : 10 000; the sex distribution (male : female) was 1.4 : 1. Holoprosencephaly was found in one dichorionic twin pregnancy and one pair of conjoined twins. Among the 30 cases of holoprosencephaly, 18 were alobar, five were semilobar, two were lobar, two were lobar variants, and three were anencephalic. The facial features varied considerably. Sixty-seven per cent (20/30) had associated structural anomalies that were not related to the cerebral and facial holoprosencephaly condition. Thirty-seven per cent (11/30) had detectable chromosome aberrations and 23% (7/30) had nonchromosomal syndromal origin. The size or shape of the head was abnormal in 83% (25/30) of holoprosencephaly cases. CONCLUSION: This study indicates that holoprosencephaly represents a heterogeneous entity with different etiologies and clinical appearances. The fact that holoprosencephaly features are found associated with particular conditions such as fronto-nasal dysplasia (2/30; 6.7%), agnathia-otocephaly (3/30; 10%), and anencephaly (3/30; 10%), suggests that these may be underreported conditions in other large holoprosencephaly series. PMID- 11851966 TI - The human ductus venosus between 13 and 17 weeks of gestation: histological and morphometric studies. AB - BACKGROUND: Doppler studies of the ductus venosus are increasingly being integrated in the assessment of fetal well-being. Establishing the precise morphology and structure of the ductus venosus would provide a better understanding of Doppler findings during fetal adaptation. There is conflicting evidence from previous studies about the structure of the ductus venosus, especially with regard to the presence of a sphincter at the ductus venosus inlet. The aim of this study was to examine the morphology and histological structure of the ductus venosus wall and surrounding tissues at 13-17 weeks' gestation. DESIGN: This was a prospective study on 28 fetuses obtained from medical termination of pregnancies between 13 and 17 weeks' gestation. Scanning electron microscopy and histological and immunohistochemical studies were carried out on ductus venosus sections obtained from different spatial planes. RESULTS: The inlet of the ductus venosus contained a shelf which was rich in elastin, but devoid of any evidence of a smooth muscle sphincter. The isthmus of the ductus venosus above the inlet was narrowed, giving the lumen of the vessel an hourglass appearance. The endothelial surface of the ductus venosus, above the level of the inlet, showed longitudinal corrugations along its entire length. Longitudinally arranged elastin fibers were also seen along the length of the ductus venosus. A single layer of longitudinally arranged smooth muscle cells was present along the entire length of the ductus venosus, with occasional individual nerve cells visible in this layer. CONCLUSIONS: The presence of an elastin-rich shelf and a narrow ductus venosus inlet orifice may act to accelerate flow from the portal sinus into a high-velocity system in the ductus venosus. The abundant elastin fibers in the adventitia of the ductus venosus may help antegrade wave propagation by elastic recoil. This study clearly demonstrates the lack of an anatomical smooth muscle sphincter at the ductus venosus inlet. However, the combination of endothelial corrugations and innervated smooth muscle support the hypothesis that the ductus venosus is an actively regulated vessel with the capacity to rapidly change diameter along its entire length in response to certain stimuli. PMID- 11851967 TI - Normal cardiac flow velocities at 14-16 weeks of gestation measured by transvaginal ultrasound. AB - OBJECTIVE: Doppler ultrasound has been used to study the flow velocities through the valves and arteries of the fetal heart. Using transvaginal sonography, we sought to determine normal values for flow velocities through the fetal heart valves at 14-16 weeks of gestation. METHODS: Eighty-seven normal fetuses were examined. The flow velocity waveforms were visualized using Doppler ultrasound with the sample volume calipers placed just distally to the four fetal valves. Peak E- and A-wave velocities were recorded for the tricuspid and mitral valves and peak systolic flow velocities were recorded for the ascending aorta and pulmonary arteries. Linear regression for comparison of flow velocities with gestational age was performed. RESULTS: There was no significant correlation between any of the velocities and gestational age. CONCLUSIONS: Flow velocities through the fetal cardiac valves remain unchanged at 14-16 weeks of gestation as measured by transvaginal Doppler sonography. PMID- 11851968 TI - Poor prenatal detection rate of cardiac anomalies in Noonan syndrome. AB - BACKGROUND: The wide variation and nonspecific nature of many of the associated ultrasonographic findings complicate prenatal diagnosis of Noonan syndrome. The aim of the present study was to define the rate of prenatal diagnosis of heart malformations in cases diagnosed postnatally with Noonan syndrome. METHODS: English-language literature review of 29 cases of Noonan syndrome examined prenatally with confirmed postnatal diagnosis and four case reports from our center. RESULTS: Cases were evaluated for cervical spine pathologies, cardiac anomalies and other pathological findings, including hydrops fetalis and polyhydramnios. Cardiac anomalies were suspected in only nine of 33 cases; three of these were associated with cystic hygroma. Cardiac anomalies were eventually diagnosed in 31/33 cases postnatally. Polyhydramnios was diagnosed in 19/33 cases in the third trimester, and hydrops fetalis was detected in eight of 33. Cystic hygroma was present in a total of nine cases at mid-trimester. CONCLUSIONS: Noonan syndrome is characterized by late-onset and progressive pathologies, particularly the associated cardiac anomalies, which develop through the course of gestation and postnatal life. This complicates or precludes prenatal diagnosis at mid-trimester or at any time in the prenatal period, and partly explains the low rate of detection of fetal cardiac lesions in this syndrome. PMID- 11851969 TI - Serial sonographic findings of four fetuses with homozygous alpha-thalassemia-1 from 21 weeks onwards. AB - OBJECTIVES: To evaluate the potential usefulness of noninvasive ultrasound assessment of fetal anemia in the diagnosis and management of fetuses with homozygous alpha-thalassemia-1. METHODS: We describe four pregnancies complicated by fetal homozygous alpha-thalassemia-1. They presented with ultrasound abnormalities before the development of hydrops. As part of evaluating the fetal condition, we performed ultrasound and Doppler studies aimed at identifying fetal anemia. These studies included evaluation of intrahepatic umbilical venous maximum flow velocity, middle cerebral artery peak flow velocity, fetal liver length and spleen perimeter. RESULTS: In all four fetuses, ultrasound and Doppler studies suggested the presence of fetal anemia. Homozygous alpha-thalassemia-1 was diagnosed in all cases, with fetal blood sampling confirming anemia in three fetuses. The majority of the intrahepatic umbilical venous maximum flow velocity and middle cerebral artery peak flow velocity measurements were above the 95th centile. Two fetuses underwent intrauterine transfusion and fetal blood flow velocities returned to normal after correction of the fetal anemia. The fetal liver length and spleen perimeter measurements showed a similar trend, although they were less consistent before 28 weeks. CONCLUSION: Non-invasive ultrasound parameters, in particular quantification of intrahepatic umbilical venous maximum flow velocity and middle cerebral artery peak flow velocity, were found to be useful in the diagnosis and management of fetal anemia in pregnancies with fetal homozygous alpha-thalassemia-1. PMID- 11851970 TI - Nuchal fold thickness at 14-16 weeks' gestation as measured by transvaginal sonography. AB - OBJECTIVE: To establish normal values of fetal nuchal fold thick-ness at 14-16 weeks of gestation by transvaginal sonography. METHODS: Transvaginal sonography was used to measure nuchal fold thickness in 182 normal pregnancies at 14-16 weeks of gestation. Nuchal fold thickness was measured as the distance from the outer skull bone to the outer skin surface in the transverse axial image in the suboccipital-bregmatic plane of the head. RESULTS: There was no significant association between nuchal fold thickness and gestational age (r = 0.084; P = 0.258). The mean (standard deviation) was 2.2 (0.5) mm and the 95th centile was 3.0 mm. CONCLUSIONS: The 95th centile of nuchal fold thickness measured by transvaginal sonography at 14-16 weeks is 3.0 mm. PMID- 11851971 TI - Value of sonomorphological criteria of the endometrium in women with postmenopausal bleeding: a multivariate analysis. AB - OBJECTIVE: This study was performed to examine whether an improvement in the transvaginal sonographic evaluation of the endometrium is possible by the addition of sonomorphological criteria to the measurement of endometrial thickness in women with postmenopausal bleeding. METHODS: Various sonomorphological criteria were analyzed prospectively in 321 patients with postmenopausal bleeding. In a logistic regression model relevant criteria were selected and a diagnostic formula for differentiation of endometrial sonographic findings was derived. RESULTS: The criteria of endometrial structure, endometrial myometrial border and endometrial thickness were significant for the differentiation of malignancy. These results allowed an estimation of the probability of malignancy for each sonographic endometrial finding. Using the cut off point of 0.1 for the probability of malignancy, the sensitivity and specificity were 96.8% and 61.9%, respectively, with an accuracy of 72.3%. In contrast, the differentiation by endometrial thickness as the sole criterion (cut off point > or = 5 mm) achieved a sensitivity of 97.9% and a specificity of 33.2%, with an accuracy of 52.3%. CONCLUSION: A useful diagnostic formula based on sonomorphological and metric criteria for endometrial differentiation was obtained. The specificity and accuracy increased with a minimal loss of sensitivity. However, estimates for sensitivity, specificity, and accuracy may be overoptimistic because they were derived from the same data used for development of the model. PMID- 11851972 TI - The reproducibility of endometrial volume acquisition and measurement with the VOCAL-imaging program. AB - OBJECTIVES: To determine the intraobserver reproducibility of volume acquisition and repeatability of endometrial volume measurements using the VOCAL-imaging program (Virtual Organ Computer-aided AnaLysis). METHODS: Ten three-dimensional (3D) ultrasound datasets of the same endometrium were obtained from one patient during one scan session. For every 3D dataset, the endometrial volume was calculated by four different measurement techniques; three rotational methods and one conventional method. A single observer acquired each dataset and conducted all measurements. For the rotational technique, the manual mode was used to trace the myometrial-endometrial border through three different predefined rotations of 30 degrees, 15 degrees and 9 degrees . Endometrial volume was also measured in a conventional manner by scrolling through one of the multiplanar images and outlining the contour in another. Ten consecutive measurements were taken for each of the four measurement techniques so that 40 endometrial volume readings were obtained for each of the 10 3D-ultrasound datasets. RESULTS: Endometrial volume measurements were more reproducible with the rotational technique. Rotational methods employing a rotation step of less than 30 degrees were associated with coefficients of variation below 1% and intraclass correlation coefficients above 0.94. Within each of the 10 different scans, the homogeneity of the variance of measurements was significantly different according to the method of measurement (P = 0.022 to < 0.001). Post-hoc tests revealed that both the conventional and 30 degrees rotational methods resulted in a significantly smaller mean endometrial volume than both the 15 degrees and the 9 degrees rotational methods. CONCLUSIONS: Endometrial volume acquisition is reproducible and endometrial volume measurements are repeatable but they depend upon the technique of volume calculation in so much as rotational methods employing a rotation step of less than 30 degrees are associated with a significantly smaller variance in measurements and a significantly greater mean endometrial volume. PMID- 11851973 TI - Reproducibility of transvaginal three-dimensional endometrial volume measurements with virtual organ computer-aided analysis (VOCAL) during ovarian stimulation. AB - OBJECTIVE: To calculate inter- and intraobserver reliabilities for three dimensional endometrial volume measurements during in vitro fertilization using virtual organ computer-aided analysis (VOCAL). PATIENTS AND METHODS: Three dimensional ultrasound measurements of the endometrium using VOCAL software were performed on the day of oocyte retrieval in each of 79 consecutive patients undergoing in vitro fertilization or intracytoplasmic sperm injection. Endometrial volume was calculated every 15 degrees and every 30 degrees by two different observers in order to determine the reproducibility of the technique. RESULTS: Intraobserver reliability for the 15 degrees measurements was 0.97 for both observers; for the 30 degrees measurements, it was 0.93 for one observer and 0.96 for the other. Interobserver reliability was 0.80 for the endometrial volumes calculated every 15 degrees and 0.83 for the volumes calculated every 30 degrees. The intra- and interobserver measurement agreement showed good reproducibility. However, the volumes calculated every 15 degrees were more accurate because the means of differences were close to zero. CONCLUSION: VOCAL provides a reproducible method for the estimation of the endometrial volume. PMID- 11851975 TI - Three-dimensional ultrasound evaluation of short-rib polydactyly syndrome type II in the second trimester: a case report. AB - Prenatal diagnosis of short-rib polydactyly syndrome is possible and has been reported in literature, but a precise ultrasound diagnosis is not easy. We report a case in which three-dimensional ultrasound was used in the evaluation of the disorder. The contribution and potential application of three-dimensional sonography in the prenatal diagnosis of short-rib polydactyly syndrome and other fetal skeletal malformations is discussed. PMID- 11851974 TI - Prospective sonographic assessment of uterine artery embolization for the treatment of fibroids. AB - OBJECTIVES: To evaluate sonographic features following uterine artery embolization and to assess using ultrasound the efficacy of embolization as the primary treatment of fibroids. DESIGN: Fifty-eight women (mean age, 44.5 years; range, 33-65 years) suffering from symptoms due to fibroids (menometrorrhagia, bulk-related symptoms, pelvic pain) were followed-up after uterine artery embolization by ultrasound examination at 3 months, 6 months, 1 year and 2 years with assessment of volume and vascularization of fibroids as well as uterine vascularization. RESULTS: Fifty-eight patients were examined at 3 months, 46 at 6 months, 36 at 1 year and 19 at 2 years. Most patients were improved or free of symptoms at 3 months (90%), 6 months (92%) and 1 year (87%) and all monitored patients were free of symptoms at 2 years. Clinical failure of treatment occurred in only two cases (3%). Progressive significant reduction in fibroid size with reference to the baseline was demonstrated during follow-up from 3 months (-29%) to 24 months (-86%). Absence of intrafibroid vessels was observed in all except three cases as early as 3 months, whereas perifibroid vessels persisted in 21 cases. No changes in uterine vascularization or uterine artery resistance were noted. CONCLUSIONS: Uterine artery embolization is a valuable endovascular method for the treatment of fibroids, resulting in marked reduction in fibroid size and disappearance of intrafibroid vessels without reduction in uterine vascularization which is well depicted by sonography. PMID- 11851976 TI - Diagnosis of skeletal dysplasia by multidisciplinary assessment: a report of two cases of thanatophoric dysplasia. AB - Skeletal dysplasias, a heterogeneous group of bone growth disorders, can be detected by routine prenatal ultrasound examination. As it is difficult to make a specific diagnosis, prediction of prognosis is of importance for obstetric management. In order to specify diagnosis, radiological, pathological and molecular genetic examination are often required. Our report describes two cases of thanatophoric dysplasia with different fetal sonographic findings. The classical classification of type I and II seems to be ambiguous as, in both cases, the same mutation in the fibroblast growth factor receptor 3 gene was found. The importance of comprehensive multidisciplinary assessment is emphasized. PMID- 11851977 TI - Sonographic appearance of appendiceal mucocele. AB - We present a case of the diagnosis of an appendiceal mucocele in a 23-year-old woman. The unusual preoperative sonographic appearance of the lesion is described and its clinical significance and differential diagnosis are discussed. PMID- 11851978 TI - Three-dimensional ultrasound in the evaluation of complex anomalies associated with fetal ventral midline defects. PMID- 11851981 TI - Nonsyndromic cleft lip and palate: complex genetics and environmental effects. AB - Nonsyndromic cleft lip and palate (CL/P) is a common craniofacial malformation with a complex genetic component. Attempts at identifying susceptibility loci via family and case-control studies have proved inconsistent. It is likely that initial predictions of the complex interactions involved in facial development were underestimated. The candidate gene list for CL/P is getting longer and the need for an impartial, systematic screening technique, to implicate or refute the inclusion of particular loci, is apparent. Large-scale multi-centre collaborations will also be necessary if we are to pursue the avenues of gene environment interactions, which have opened up over recent years. So we are faced with the question 'Can this complex trait be too complex?' PMID- 11851982 TI - Exclusion of 5 functional candidate genes for distal hereditary motor neuropathy type II (distal HMN II) linked to 12q24.3. AB - Distal hereditary motor neuropathies (distal HMNs) are characterised by degeneration of anterior horn cells of the spinal cord resulting in muscle weakness and atrophy. Distal HMN type II is genetically linked to chromosome 12q24.3 and located within a 13 cM region flanked by markers D12S86 and D12S340. We previously excluded the human phospholipase A2 group 1B gene (PLA2G1B) as the disease causing gene. Here, we report the mutation analysis of five other candidate genes localised within the distal HMN II region: the cytoskeletal proteins paxillin (PXN) and restin (RSN); the acidic ribosomal phosphoprotein, large P0 subunit (RPLP0); a nucleoside diphosphate kinase (NME2B); and the beta 3 subunit of the voltage-gated calcium channel (CACNB3). DNA sequencing of the coding regions was performed but no disease causing mutations could be identified, hence excluding these five genes for distal HMN type II. PMID- 11851983 TI - Angiotensin converting enzyme insertion allele in relation to high altitude adaptation. AB - Angiotensin converting enzyme (ACE) gene I/D polymorphism has been associated with high altitude (HA) disorders as well as physical performance. We, however, envisage that the polymorphism may be associated with adaptation to the hypobaric hypoxia of altitude, thus facilitating physical performance. For this purpose, three unrelated adult male groups, namely (1) the Ladakhis (HLs), who reside at and above a height of 3600 m, (2) lowlanders, who migrated to Ladakh (MLLs), and (3) resident lowlanders (LLs), have been investigated. The HLs had significantly (p & 0.001) greater numbers of the II homozygotes and the ID heterozygotes than the DD homozygotes, the genotype distribution being 0.46, 0.43 and 0.11 for II, ID and DD genotypes respectively. The MLLs comprised 60% II homozygotes, which was higher (p & 0.001) than the HLs (46%). In the LLs, the heterozygotes were greater (p & 0.001) in number than the II and DD homozygotes. The I allele frequency was 0.72 in the MLLs, 0.67 in the HLs and 0.55 in the LLs. Polymorphism study suggested that the II genotype could be associated with altitude adaptation, which might influence physical efficiency. PMID- 11851984 TI - Gm and Km alleles in two Spanish Pyrenean populations (Andorra and Pallars Sobira): a review of Gm variation in the Western Mediterranean basin. AB - Two Spanish eastern Pyrenean populations, Andorra and Pallars Sobira, have been tested for G1m(1,2,3,17), G2m(23), G3m(5,6,10,11,13,14,15,16,21,24,28) and Km(1) immunoglobulin allotypes. Km allele and Gm haplotype frequencies in both samples fit well into the Western Mediterranean and, more strictly, Pyrenean ranges with some peculiarities: Andorra showed an elevated frequency (14.7%) of the typical Asian and European Gm21,28;1,2,17;. haplotype, while Pallars Sobira was characterized by high values (3.7%) of Gm5*;1,17;., a typical sub-Saharan Gm haplotype. Gm diversity assessed through genetic distance and variance analyses revealed a significant geographic partition (4.3%) of Mediterraneans among south, north-east, and north-west groups. It is interesting to note the relatively low genetic variance (2.1%) found between south and north-western Mediterraneans that could reflect ancient population relationships. More locally, genetic boundaries and diversity analyses failed to indicate any geographic pattern and/or genetic differentiation related with the political border in the Pyrenees. The present pattern of variation in this area is probably the result of genetic isolation processes, in addition to some specific demographic phenomena, in the Pyrenean valleys. PMID- 11851985 TI - Phylogeography of the human mitochondrial haplogroup L3e: a snapshot of African prehistory and Atlantic slave trade. AB - The mtDNA haplogroup L3e, which is identified by the restriction site +2349 MboI within the Afro-Eurasian superhaplogroup L3 (-3592 HpaI), is omnipresent in Africa but virtually absent in Eurasia (except for neighbouring areas with limited genetic exchange). L3e was hitherto poorly characterised in terms of HVS I motifs, as the ancestral HVS-I type of L3e cannot be distinguished from the putative HVS-I ancestor of the entire L3 (differing from the CRS by a transition at np 16223). An MboI screening at np 2349 of a large number of Brazilian and Caribbean mtDNAs (encompassing numerous mtDNAs of African ancestry), now reveals that L3e is subdivided into four principal clades, each characterised by a single mutation in HVS-I, with additional support coming from HVS-II and partial RFLP analysis. The apparently oldest of these clades (transition at np 16327) occurs mainly in central Africa and was probably carried to southern Africa with the Bantu expansion(s). The most frequent clade (transition at np 16320) testifies to a pronounced expansion event in the mid-Holocene and seems to be prominent in many Bantu groups from all of Africa. In contrast, one clade (transition at np 16264) is essentially restricted to Atlantic western Africa (including Cabo Verde). We propose a tentative L3e phylogeny that is based on 197 HVS-I sequences. We conclude that haplogroup L3e originated in central or eastern Africa about 46,000 (+/-14,000) years ago, and was a hitchhiker of much later dispersal and local expansion events, with the rise of food production and iron smelting. Enforced migration of African slaves to the Americas translocated L3e mitochondria, the descendants of which in Brazil and the Caribbean still reflect their different regional African ancestries. PMID- 11851986 TI - Epistasis and its possible effects on transmission disequilibrium tests. AB - Most approaches to analysing complex genetic disorders are based on an underlying model that assumes that the probability of being affected is mainly due to the effect of a major, single disease locus. Where the penetrance (the conditional probability of being affected) is hypothesised to be influenced by known covariates, methods have been developed to accommodate these covariates appropriately in the analysis. Unexplained variation in the incomplete penetrance is then attributed to either unspecified environmental effects or to other genes of small effect that might act epistatically with the 'disease gene'. Recently, there have been some proposals to incorporate two (or more) possibly epistatic identified loci as major effects in the analysis. However, the effect of two (or more) major epistatic disease genes on the analysis of data that assumes a single 'disease gene', and so ignores any other major gene, does not seem to have been explored. This investigation is undertaken here for triad data consisting of affected singletons and their parents. Following development of a quite general underlying genetic model, a global approach to analysing two marker loci for triad data is developed. This approach is the appropriate one for such data where two, possibly epistatic, loci have been identified a priori. The motivation for the development of this methodology is to evaluate the possible effect a second epistatic disease locus might be having on the results for triad data collected and analysed in different population studies, where the underlying analysis is based on an underlying major, single disease locus model. It is shown that, dependent on the population parameters associated with the other unidentified major disease gene, results concerning the single 'disease gene' can vary markedly. So, finding that conclusions differ from study to study may be indicative of the 'disease gene' under investigation acting epistatically with other major disease genes. PMID- 11851987 TI - Correcting for multiple analyses in genomewide linkage studies. AB - The dissection of complex traits frequently calls for multiple analyses to be performed, including the use of both multiple phenotypes and genetic models. These multiple phenotypes and models are often not independent, and hence the necessary correction for the multiple testing is not straightforward. In this paper we offer a new approach to address the problem of how to correct for non independent multiple analyses in genomewide linkage studies. We describe one method of how to determine the number of 'effectively independent' tests performed in a linkage study using simple linear regression techniques. Further we describe how to use such information to establish genomewide significance thresholds for infinitely dense genomewide maps. PMID- 11851988 TI - Power of regression and maximum likelihood methods to map QTL from sib-pair and DZ twin data. AB - A common study design to map quantitative trait loci (QTL) is to compare the phenotypes and marker genotypes of two or more siblings in a sample of unrelated sib groups, and to test for linkage between chromosome location and quantitative trait values. The simplest case is sib pairs only, in particular dizygotic twin pairs, and a simple and elegant regression method was proposed by Haseman & Elston in 1972 to test for linkage. Since then, several other methods have been proposed to test for linkage. In this study, we derived the statistical power of linear regression and maximum likelihood methods to map QTL from sib pair data analytically, and determined which methods are superior under which set of population parameters. In particular, we considered four regression-based and three maximum likelihood-based approaches, and derived asymptotic approximations of the mean test statistic and statistical power for each method. It was found, both analytically and by computer simulation, that the revisited or new Haseman Elston method (based upon the mean-corrected crossproduct of the observations on sib-pairs) is less powerful than a full maximum likelihood approach and is also inferior to the Haseman-Elston method under a realistic range of values for the population parameters. We found that a simple regression method, based upon both the squared difference and the mean-corrected squared sum of the observations on sib-pairs, is as powerful as a full maximum likelihood approach. Our derivations of statistical power for regression and maximum likelihood methods provide a simple way to compare alternative methods and obviate the need to perform elaborate computer simulations. DZ twin pairs are likely to be more powerful for linkage analysis than ordinary siblings because they may share more common environmental effects, thereby increasing the proportion of within-family variance that is explained by a QTL. PMID- 11851990 TI - [Locoregional recurrences after conservative treatment of breast cancer stage I II]. AB - BACKGROUND: Since 1992 conservative treatment of breast cancer (stage I and II: clinical TNM) has been established as an alternative to mastectomy in our hospital. The aim of this retrospective study was to analyse locoregional recurrence features and to compare prognosis with regard to to site of recurrence. METHOD: Between 1987 and 1993, 489 patients with breast cancer (stages I and II) were treated with conservative surgery and radiation therapy at the Hospital Sant Pau of Barcelona. Mean follow-up was 58.8 months [between 12 144]. 35 patients developed locoregional recurrence. We considered two groups: local recurrence in breast only; and locoregional recurrence such as nodal recurrence with or without simultaneous breast recurrence. Diagnosis was confirmed by histopathologic analysis. An extensive study was performed in all patients to rule out distant metastasis. Last follow-up was December 1999. RESULTS: The locoregional recurrence rate after conservative treatment was 7.5% and that of local recurrence was 3.06%. Recurrences were diagnosed in 80% of patients by physical examination, while 20% of patients had noticed the tumor recurrence themselves. Histologic grade III tumors had a higher number of locoregional recurrences than local recurrences (p = 0.030). Locoregional recurrences had lower overall survival rate (p = 0.0005), lower disease-free survival rate (p = 0.0012) and shorter time period without distant metastasis (p < 0.0005) than local recurrences. CONCLUSIONS: Most recurrences were diagnosed by clinical examination during follow up. Histologic grade III was related to locoregional recurrences. Local recurrences had a better prognosis than locoregional recurrences. PMID- 11851991 TI - [Meta-analysis of the scientific evidence on the usefulness of sporadic intake of acetylsalicylic acid in the prevention of coronary heart disease]. AB - BACKGROUND: The present study was aimed at determining whether the sporadic intake of acetylsalicylic acid (ASA) shows a protective effect on the appearance or attenuation of coronary disease events. METHODS: The analysis was based on articles found in EMBASE, MEDLINE and the Cochrane Library. Scientific rigour was further assessed. We looked for original articles with clinical trial, cohorts, and case-control or cross-sectional study designs, where the effect could be assessed by the odds ratio (OR). RESULTS: A meta-analysis showed a protective effect of sporadic ASA intake on the prevention of acute myocardial infarction (OR for fixed effects = 0.75, CI 95%, 0.63-0.88, p < 0.0006), which was more important in men than in women, and on the prevention of cardiovascular mortality (OR for fixed effects = 0.61, CI 95%, 0.59-0.64, p < 0.0001). However, overall mortality was found to be higher in those groups receiving the drug (OR for fixed effects = 1.20, CI 95%, 1.05-1.37, p = 0.0006). None of these effects was significant when performing a random effect analysis. ASA also attenuated acute coronary syndromes (OR for fixed effects = 0.34, CI 95%, 0.26-0.45, p < 0.0001). CONCLUSIONS: These results suggest that the sporadic intake of ASA may have a protective particularly in men and attenuating effect on acute myocardial infarction, in addition to playing a role in preventing cardiovascular mortality but not overall mortality. Further studies to confirm these effects are warranted. PMID- 11851992 TI - [Do not resuscitate orders in patients with acute stroke. What circumstances decide their use?]. AB - BACKGROUND: Patients with acute stroke are known to have a poor prognosis after cardiopulmonary resuscitation manoeuvres (CPR), and their application should be revisited in these patients. Although clinical criteria for a 'do not resuscitate order' (DNR) are available in some countries, studies about DNR in stroke are lacking in Spain. The purpose of this study was to evaluate the frequency of DNR orders in patients with acute stroke and to identify factors influencing decision making in them. PATIENTS AND METHOD: All patients with acute stroke who had cardiac and pulmonary arrest were prospectively included in the study during one year. Clinical and demographic data of patients and data related to doctors were recorded and analysed. RESULTS: 165 patients had a cardiac and pulmonary arrest and 17 (10%) of them had had a DNR order. No factor was significantly associated with DNR decision-making. CONCLUSIONS: DNR orders were scarcely applied and explicit clinical criteria for their application were lacking. It is necessary to implement DNR policies in Spain in order to improve the use of CPR manoeuvres in patients with acute stroke. PMID- 11851993 TI - [Promotion of the blood donation in a hospital: comparison of two approaches]. AB - BACKGROUND: Our purpose was to evaluate the results of the promotion of blood donation in patients undergoing stem cell transplantation (SCT) and to compare it with this promotion in remaining in-patients. PATIENTS AND METHODS: The hematologist and the blood bank staff informed the patient undergoing SCT about the need for transfusions. Donors were selected for blood or platelet donation. In remaining in-patients, promotion was performed by the blood bank staff by interviewing patients. RESULTS: Between January 2000 and May 2001, 48 SCT patients were included: 41 (85%) yielded 453 possible donors. Of them, 306 donated blood, 77 platelets and 82 were excluded. The number of donations was 484 (389 whole blood and 105 platelets), and 25% (n = 98) of patients donated blood subsequently. Among in-patients 1,950 interviews were carried out: 533 (27.3%) yielded 901 donors (p < 0.001). Overall, 1,146 blood donations were obtained and 15% (135) of these donors donated blood subsequently (p < 0.001). CONCLUSION: Collaboration of the hematologist in the promotion of blood donation leads to an increase in the rate and frequency of donations. PMID- 11851994 TI - [Inflammation and cardiac disease]. PMID- 11851995 TI - [Clinical study of 17 patients with cheiro-oral-pedal syndrome]. AB - BACKGROUND: To characterize the clinical features, etiology and prognosis in cheiro-oral-pedal syndrome. PATIENTS AND METHOD: Descriptive study of 17 patients with cheiro-oral-pedal syndrome included in the Sagrat Cor Hospital of Barcelona Stroke Registry over a 11 year period. RESULTS: Cheiro-oral-pedal syndrome was present in 6 patients (35%), cheiro-oral syndrome in 10 (59%) and isolated oral syndrome in 1 (6%). Cheiro-oral-pedal syndrome was caused by a lacunar infarct in 88% of patients and by atherotrombotic infarcts in 12%. Cheiro-oral-pedal syndrome accounted for 0.7% of all acute strokes (n = 2.244), 1% of all cerebral infarcts (n = 1.649), 3.5% of all lacunar infarcts (n = 422) and 18.5% of all pure sensory lacunar infarcts (n = 80). Thalamus (n = 13), internal capsule (n = 2), striatocapsular involvement (n = 1) and fronto-parietal involvement (n = 1) were the cerebral topographies. Absence of in hospital mortality and absence or mild neurological deficit at discharge from the hospital were present in all the patients. CONCLUSIONS: Lacune hypothesis is present in cheiro-oral-pedal syndrome. Cheiro-oral-pedal syndrome was caused by a lacunar infarct in 88% of patients. The thalamic topography is the most frequent and the prognosis in cheiro-oral-pedal syndrome is good. PMID- 11851996 TI - [Back to home]. PMID- 11851997 TI - [Mental health in immigrants: the new challenge]. PMID- 11851998 TI - [Double-blind. The bias control in the performance of clinical trials. Contradictions, lacks and implications]. PMID- 11851999 TI - [Sperm quality and hormonal levels in oligozoospermic males treated with follicle stimulating hormone]. PMID- 11852000 TI - [Rapid whole blood test for the diagnosis of H. pylori infection. Can it be recommended for the diagnosis of the infection in clinical practice?]. PMID- 11852001 TI - [HELLP syndrome]. PMID- 11852003 TI - [Hyperammonaemic coma after valproate therapy as adult onset of ornithine transcarbamylase deficiency]. PMID- 11852004 TI - [Coronary artery surgery in the first 24 hours after myocardial infarction]. AB - BACKGROUND: Thrombolysis and angioplasty in the first hours after myocardial infarction minimize necrosis, leading to better early and late survival, but these therapies have limited effect in patients with three-vessel disease and cardiogenic shock. Emergency coronary surgery is an alternative treatment in some cases. AIM: To assess perioperative complications, mortality and long-term survival in patients undergoing coronary surgery within 24 h of myocardial infarction. PATIENTS AND METHODS: We retrospectively studied 57 patients undergoing surgery within 24 h of the onset of symptoms of myocardial infarction between 1982 and 1998. Multiple vessel disease was present in 31 patients (54%), shock or cardiac arrest in 19 (33%) and coronary angiography complications in 7 (12%). The mean time between onset of symptoms and surgery was 6.32 h. At the beginning of surgery 32 patients (56%) were hemodynamically stable, 15 (26%) were in shock and 10 (17%) were in cardiac arrest. RESULTS: The operative mortality was 0% for those who were hemodynamically stable at the start of surgery and 44% (11 of 25 patients) for those in shock or cardiac arrest. Shock or prior cardiac arrest were associated with higher rates of sternal infection and heart failure and longer hospital stays.Follow-up (mean 67 months) was possible for all remaining patients. The 5- and 10-year survival rates were 89 and 82%, respectively, for patients who were hemodynamically stable at the time of surgery. Five-year survival was 55%, however, for those who underwent surgery in shock or cardiac arrest. The overall rate of freedom from myocardial infarction, angioplasty or reoperation was over 95% at 5 years and over 85% at 10 years of follow-up. Age and shock or cardiac arrest were risk factors for a poor long-term outcome. CONCLUSION: The early and long-term outcome of coronary surgery within 24 h of myocardial infarction is good for patients who are hemodynamically stable when surgery begins. Shock and cardiac arrest are important risk factors for complication and death. Coronary artery bypass grafting is a good treatment option in the first hours after myocardial infarction. PMID- 11852005 TI - [Chest pain units. Organization and protocol for the diagnosis of acute coronary syndromes]. AB - The two main goals of chest pain units are the early, accurate diagnosis of acute coronary syndromes and the rapid, efficient recognition of low-risk patients who do not need hospital admission. Many clinical, practical, and economic reasons support the establishment of such units. Patients with chest pain account for a substantial proportion of emergency room turnover and their care is still far from optimal: 8% of patients sent home are later diagnosed of acute coronary syndrome and 60% of admissions for chest pain eventually prove to have been unnecessary.We present a systematic approach to create and manage a chest pain unit employing specialists headed by a cardiologist. The unit may be functional or located in a separate area of the emergency room. Initial triage is based on the clinical characteristics, the ECG and biomarkers of myocardial infarct. Risk stratification in the second phase selects patients to be admitted to the chest pain unit for 6-12 h. Finally, we propose treadmill testing before discharge to rule out the presence of acute myocardial ischemia or damage in patients with negative biomarkers and non-diagnostic serial ECGs. PMID- 11852006 TI - [Clinical studies reported in the 74th annual scientific sessions of the American Heart Association (Anaheim November 11-14, 2001)]. PMID- 11852007 TI - [Regulation of myocardial gene expression during heart development]. AB - The heart is an organ with special significance in medicine and developmental biology. The development of the heart and its vessels during embryogenesis is the result of numerous and complex processes. At present, our understanding is based on decades of meticulous anatomical studies. However, the spectacular progress of modern molecular biology and developmental biology has marked the beginning of a new era in embryology. The molecular bases for cardiogenesis are just emerging. Several families of genes with restricted expression to the heart have been identified in the last years, including genes encoding for contractile proteins, ion channels as well as transcription factors involved in tissue specific gene expression. Likewise, the analyses of regulatory elements have increased our understanding of the molecular mechanisms directing gene expression. In this review, we illustrate the different patterns of gene and transgene expression in the developing myocardium. These data demonstrate that the wide molecular heterogeneity observed in the developing myocardium is not restricted to embryogenesis but it also remains in the adulthood. Therefore, such molecular diversity should be taken into account on the design of future gene therapy approaches, having thus direct clinical implications. PMID- 11852008 TI - [Pneumopericardium complicating bronchogenic carcinoma]. PMID- 11852009 TI - [Eptifibatide blocks the increase in C-reactive protein concentration after coronary angioplasty]. AB - We measured C-reactive protein concentrations in 31 patients with interventional procedures after blocking thrombosis in 17 of them by the administration of a 12 hour long infusion of eptifibatide in order to evaluate the role of arterial thrombosis. There were no differences in C reactive protein concentration between the treated and control group pre-angioplasty (0.32 0.4 vs 0.56 0.57 md/dl; p = NS), nor post-angioplasty (0.35 0.42 vs 0.53 0.5 mg/dl, p = Ns). The eptifibatide group maintained basal C reactive protein concentrations 6 hours after the procedure, while the control group had a significant increase (0.43 0.5 vs 1.02 0.89 mg/dl; p < 0.05). There was a decrease in C reactive protein 24 hours after angioplasty in eptifibatide group (0.24 0.27 vs 1.34 0.89 mg/dl; p < 0.001), but it increased again 48 hours after the procedure although to a lesser extent than in the control group (0.57 0.55 vs 2.18 2.1 mg/dl; p < 0.05). Eptifibatide, a synthetic peptide which is a selective blocker of the platelet GP IIb/IIIa receptor significantly reduced C-reactive protein concentration after angioplasty. PMID- 11852010 TI - [Intraluminal dilation of inferior vena cava stenosis after repair of the scimitar syndrome in an adult patient]. AB - A 39 year-old woman diagnosed with anomalous drainage of middle and lower right pulmonary veins to the inferior vena cava was corrected surgically by means of baffle with patch up to the left atrium. Early after the operation the patient related intolerance to small efforts and an episode of syncope. The cardiac catheterization demonstrated the presence of a severe stenosis in the inferior vena cava, in its union with the right atrium, that was successfully treated by means of intraluminal percutaneous dilation with a catheter of Inoue. After the procedure the gradient decreased and she improved tolerance to effort, which persisted 10 months later. PMID- 11852011 TI - [Incessant ventricular tachycardia as a manifestation of myocardial ischemia]. AB - We describe four patients with incessant ventricular tachycardia after the acute phase of a myocardial infarction. Two of them had a slow heart rate, and myocardial revascularization resolved the arrhythmia after ischemia was demonstrated. In the other two cases, very fast tachycardias were interrupted by means of intravenous verapamil and clinical stabilization was achieved after failure of amiodarone and lidocaine. In one of them, revascularization prevented new recurrences, but it was not feasible in the second patient, who developed new arrhythmias. The possible mechanisms of these tachycardias and their clinical and therapeutic implications are discussed. PMID- 11852012 TI - [Myocardial ischemia caused by an anomalous circumflex coronary artery]. AB - The case of a 71-year-old male patient, with symptoms of dizzines and atypical chest pain and a positive isotopic exercise stress test, is reported. Coronary angiography demostrated an anomalous origin of the left circumflex coronary artery from right coronary ostium but no obstructive atherosclerotic coronary lesions. The possible relation between the congenital coronary anomaly and the clinical manifestations of the patient is discussed. PMID- 11852013 TI - [The mitral ring contraction and the mechanics of the ascendant segment of the ventricular myocardial band]. PMID- 11852015 TI - [Mitral valvuloplasty during pregnancy]. PMID- 11852017 TI - [MADIT II prematurely terminated: major impact on health economics?]. PMID- 11852018 TI - [Genotypes of the renin-angiotensin-aldosterone system: on the search of cardiovascular diseases]. PMID- 11852019 TI - [The involvement of the renin-angiotensin system gene polymorphisms in coronary heart disease]. AB - INTRODUCTION AND OBJECTIVES: Previous studies angiotensin-converting enzyme gene insertion/deletion polymorphism ACE (I/D), angiotensinogen gene polymorphism, and angiotensin II AT1 receptor polymorphism in relation to coronary heart disease controversial results. This study was designed to analyze the association between these gene polymorphisms and the first coronary event in individuals residing on Grand Canary Island, Spain. PATIENTS AND METHOD: Case-control study. Case subjects (n = 304) were recruited at the first coronary event; age-matched controls (n = 315) were randomly selected from the Grand Canary population. Participants were examined for the usual risk factors. Blood samples were obtained for biochemical analyses and DNA extraction. Genotyping was performed by PCR and restriction analysis. RESULTS: Neither ACE (I/D) nor AT1 receptor polymorphism was associated with coronary heart disease, whereas the frequency distribution of AGT M235T genotypes among patients and control subjects (TT: 29% and 19%; MT: 48% and 50%; MM: 22% and 31%, respectively) was statistically different (p = 0.003). Multiple logistic regression analysis identified the TT genotype of the angiotensinogen gene (OR = 1.9; 95% CI 1.1-3.4), diabetes (OR = 4.4; 95% CI 2.0-9.4) and hypertension (OR = 2.1; 95% CI 1.3-3.3) as risk factors predicting the coronary event. CONCLUSIONS: Our results provide no evidence of an association between ACE (I/D) or AT1 receptor polymorphism and coronary heart disease. However, homozygosity for the T allele of the angiotensinogen gene, diabetes and hypertension independently place individuals at higher risk of experiencing a coronary event on Grand Canary Island. PMID- 11852020 TI - [Is troponin I useful for predicting in-hospital risk for unstable angina patients in a community hospital? Results of a prospective study]. AB - INTRODUCTION AND OBJECTIVES: Before including troponin I detection in the daily practice of our hospital we performed a prospective study to determine its real usefulness and to establish the best cut-off point. METHODS: We studied 82 consecutive patients admitted with unstable angina to a community hospital. Troponin I was determined (> 10 h after chest pain). Patients were referred to a tertiary hospital for catheterization/revascularization if clinical events developed. RESULTS: Twenty-five patients (31%) suffered events during admission: recurrent angina in 23 cases (28%); heart failure in 5 (6%); exitus in 3 (4%); myocardial infarction in 1 (1%). The cut-off point for troponin I that best predicted events was 0.1 ng/ml. Patients with troponin I > 0.1 (34 patients, 42%) experienced more events [47 vs. 19%; OR = 3.8 (1.4-10.4); p = 0.01] and had higher rates of recurrent angina (42 vs. 19%), heart failure (12 vs. 2%) and exitus (9 vs 0%). Patients with ECG changes and troponin I > 0.1 showed a significantly higher percentage of events (63%) than those with ECG changes alone (23%) or troponin I > 0.1 alone (15%) or those without ECG changes and troponin I < 0.1 (17%) (p < 0.0001). CONCLUSIONS: Troponin I elevation is useful for predicting in-hospital risk for unstable angina patients admitted to a community hospital. A low cut-off value (0.1 ng/ml) predicts events. The association of ECG changes and high troponin I identifies a population at very high risk; however, the absence of both variables in patients with a diagnosis of unstable angina does not preclude the development of events. PMID- 11852021 TI - [Utility of cardiac event recorders in diagnosing arrhythmic etiology of palpitations in patients without structural heart disease]. AB - OBJECTIVE: To determine the diagnostic yield of transtelephonic event monitors for identifying the reason for palpitations in patients with no structural heart disease. PATIENTS AND METHOD: For 20 months we enrolled all patients reporting palpitations in whom heart disease had been ruled out by medical history, physical examination, ECG and transthoracic echocardiography. All patients underwent 24 h Holter monitoring, which did not provide diagnostic information. Later, a Cardiotest 4DM transtelephonic event monitor was provided to each patient for a mean of 15 3 days. We used SPSS V 10 for statistical analysis. RESULTS: Two hundred twenty-seven consecutive patients were enrolled. Mean age was 45 3 years (range 12-85); 167 were females (74%). Two hundred twelve of the 227 patients (93.3%) experienced palpitations while wearing the device, and 210 (92.5%) used the monitor correctly, recording the cardiac rhythm during palpitations. Fifteen patients (6.6%) had no palpitations during the days of study. In 125 (55%) the Cardiotest 4DM correctly recorded and transmitted arrhythmia that justified the patients' reference to palpitations. In 35 (15.4%) significant arrhythmia was detected: paroxysmal supraventricular tachycardia in 21, atrial fibrillation in 9, atrial flutter in 5, runs of ventricular extrasystoles in 4 and right outflow tract ventricular tachycardia in 1. Sinus rhythm was recorded during palpitations in 85 patients (37%), and arrhythmia as the cause could be ruled out. CONCLUSIONS: Cardiotest 4DM identifies arrhythmia in a very high proportion of patients with palpitations and no structural heart disease. PMID- 11852022 TI - [The efficacy of scheduled cardioversion in atrial fibrillation. Comparison of two schemes of treatment: electrical versus pharmacological cardioversion]. AB - INTRODUCTION AND OBJECTIVES: Atrial fibrillation is an arrhythmia with high morbidity and mortality. Restoring sinus rhythm is one of the principle objectives in its management. The present study aimed to assess the efficacy of scheduled cardioversion on atrial fibrillation by comparing two different therapeutic approaches: electrical vs. pharmacological cardioversion. PATIENTS AND METHOD: Two hundred thirty patients with atrial fibrillation of more than 48 hours duration and requiring sinus rhythm restoration were included. One hundred forty-four patients underwent external electrical cardioversion and 86 patients received quinidine. We analyzed the rate of success, duration of hospital stay, complications and clinical and echocardiographic variable that might predict success. RESULTS: Sinus rhythm was restored in 181 of 230 patients (79%). The rate of success was 77% (111/144 patients) in the electrical group and 81% (70 of 86 patients) in the pharmacological group (ns). In 13 pharmacological group patients for whom the first attempt failed attempt, a second attempt with electrical cardioversion was made and was successful in 8 patients (61%). No embolic complication was recorded and only two electrical disturbances were seen. Only atrial fibrillation lasting less than 8 weeks was associated with a higher success rate (p < 0.01). CONCLUSIONS: Scheduled cardioversion in atrial fibrillation is an effective technique with a high success rate and a very low rate of complication. Electrical cardioversion and pharmacological cardioversion with quinidine are similarly effective, although the latter involves a longer hospital stay. PMID- 11852023 TI - [Stress echocardiography in the pre-operative evaluation of patients undergoing major vascular surgery. Are results comparable with dypiridamole versus dobutamine stress echo?]. AB - INTRODUCTION: [corrected] Perioperative cardiovascular complications are an important cause of post-surgical morbility and mortality in patients undergoing major vascular surgery. Dobutamine Stress Echo is considered one of the methods of choice in the detection of coronary artery disease in this subgroup of patients. OBJECTIVES: . Our aim was to analyze if dipyridamole stress echocardiography could be used as an alternative to Dobutamine Stress Echo in the perioperative evaluation of patients in need of major vascular surgery. PATIENTS AND METHOD: The result of consecutives dypiridamole and dobutamine stress exams prior to vascular surgery were reviewed. We analyzed if those patients with a positive stress echo presented a higher number of cardiac events during and after surgery than those with negative stress echo. The negative and positive predictive values were calculated for both techniques. RESULTS: 133 stress exams were analysed: 39 with dobutamine and 94 with dipyridamole. Of the 39 dobutamine studies 2 were positive, 29 negatives and 8 non conclusive. Of the 94 dypiridamole studies 13 were positive and 81 negatives. None of the patients with a positive dobutamine echo underwent surgery. The negative predictive value for dobutamine echo was 96.5%, quite similar to that of dypiridamole stress echo (97.5%). CONCLUSION: Dipyridamole stress echocardiography is a valid alternative to dobutamine echocardiography in the pre-surgical evaluation of patients undergoing major vascular surgery. PMID- 11852024 TI - [Predictive factors of abnormal dynamic intraventricular gradient after valve replacement in severe aortic stenosis]. AB - INTRODUCTION: Dynamic intraventricular gradients (DIG) after valve replacement in severe aortic stenosis have been reported, although the incidence of DIG and clinical signs are still poorly understood.Aim. To evaluate the incidence of DIG)and determine risk factors and associated morbimortality. PATIENTS AND METHOD: One hundred nine consecutive patients with severe aortic valve stenosis undergoing valve replacement were studied prospectively by echocardiography to detect the postoperative appearance of DIG, defined as a maximum flow velocity >/= 2.5 m/s. RESULTS: Sixteen patients (14.9%) developed postoperative DIG. Significant differences between the patients with or without DIG were found for ventricular diameter (left end-diastolic ventricular diameter (LEDVD) 43.2 vs. 47.7 mm, respectively, p < 0.001; left end-systolic ventricular diameter (LESVD) 21 vs. 29 mm, p < 0.001); left ventricular mass index (165 vs. 193 g/m(2), p < 0.05); mean aortic valve gradient (68 vs. 59 mmHg, p < 0.01),; ejection fraction (73 vs. 61%, p < 0.001). No significant differences were found with respect to ventricular wall thicknesses (septal 16.3 vs. 15.7; posterior 14.37 vs. 14.62), the presence of aortic insufficiency, or other postoperative factors (anemia, inotropic agents, etc.). CONCLUSIONS: DIG after aortic valve replacement to treat severe stenosis is not unusual (15%). DIG is usually found at a midventricular location, close to the septum. In patients with postoperative DIG the most common associated factors were small LEDVD, high ejection fractions and ratios of intraventricular septal to posterior wall ratios, high valve gradients and small left ventricular masses. Preoperative echocardiography can identify patients with a higher risk of developing DIG after aortic valve replacement. PMID- 11852025 TI - Reversal of sex differences in morphine analgesia elicited from the ventrolateral periaqueductal gray in rats by neonatal hormone manipulations. AB - Male rats display significantly greater analgesic responses than female rats following systemic, ventricular and intracerebral morphine administration into either the ventrolateral periaqueductal gray (vlPAG) or the rostral ventromedial medulla, and following beta-endorphin administration into the vlPAG. Although adult gonadectomy severely reduces nonopioid forms of swim stress-induced analgesia, the marked sex differences in morphine analgesia were minimally affected by either male or female adult gonadectomy. Since very little is known about neonatal effects of gonadal hormones upon sex differences in morphine analgesia elicited from the vlPAG, the present study evaluated the effects of neonatal (within 1 day of birth) castration in male rat pups relative to sham operated controls, and systemic androgenization with testosterone propionate in female rat pups relative to vehicle-injected controls upon baseline nociceptive thresholds and morphine analgesia elicited from the vlPAG in rats tested as adults. Significant sex differences in morphine analgesia elicited from the vlPAG were observed with adult males receiving neonatal sham surgeries displaying significantly greater morphine analgesia on two nociceptive measures than adult females tested during the estrous phase and receiving neonatal vehicle injections. Neonatal gonadectomy essentially reversed the pattern of sex difference effects upon morphine analgesia elicited from the vlPAG. Neonatally castrated male rats tested in adulthood displayed dramatic reductions in morphine analgesia elicited from the vlPAG on both the tail-flick (approximately 15-fold rightward shift) and jump (6-fold rightward shift) tests relative to sham operated males, and essentially mirrored those of vehicle-treated females. Conversely, neonatally-androgenized female rats tested in adulthood displayed dramatic increases in morphine analgesia elicited from the vlPAG on the tail flick (5-fold leftward shift) and jump (12-fold leftward shift) tests relative to vehicle-treated females, and approximated those observed in sham-operated males. The potent differences between neonatally-castrated and sham-operated male rats and between neonatally-androgenized and vehicle-treated female rats suggest a possible 'organizational' role of gonadal hormones in mediating sex differences in morphine analgesia elicited from the vlPAG. PMID- 11852026 TI - Oxytocin stimulates cerebral blood flow in rainbow trout (Oncorhynchus mykiss) through a nitric oxide dependent mechanism. AB - Our knowledge of the regulation of cerebral blood flow (CBF) in ectothermic vertebrates is still very limited. In endothermic vertebrates several peptides have been shown to affect CBF through nitric oxide (NO) dependent mechanisms. Using epi-illumination microscopy in rainbow trout in vivo, we have examined the effects of topically administered oxytocin, arginine vasopressin, substance P and bradykinin on the CBF (measured as erythrocyte velocity in venules on the optic lobes). Of these peptides, only oxytocin induced a dose dependent increase in CBF velocity. Blood pressure remained unchanged and the effect was suppressed by the NOS inhibitor, N(omega)-nitro-L-arginine. This indicates that oxytocin causes NO mediated vasodilation in rainbow trout brain. PMID- 11852027 TI - Effects of excitotoxic lesions of the basolateral amygdala on cocaine-seeking behavior and cocaine conditioned place preference in rats. AB - Incentive motivation for cocaine, elicited by cocaine-associated stimuli, is thought to be involved in craving and relapse. To examine the role of the basolateral amygdala complex (BLC) in this phenomenon, we assessed the effects of post-training BLC lesions on extinction of cocaine-seeking behavior and cocaine conditioned place preference (CPP) and the effects of pre-training BLC lesions on acquisition of cocaine-CPP. In Experiment 1, rats were first trained to self administer cocaine and then received bilateral infusions of the excitotoxin, N methyl-D-aspartic acid (NMDA, 0.12 M; 0.3 microl/side), or vehicle into the BLC. They were then tested repeatedly for extinction of cocaine-seeking behavior (i.e. nonreinforced responses in the presence of cocaine-paired stimuli). Subsequently, they were trained and tested for acquisition of cocaine-CPP (i.e. increased time spent in a previously cocaine-paired, relative to a saline-paired, environment). Locomotion and compartment entries were also measured. In Experiment 2, rats were first trained and tested for cocaine-CPP, and then received NMDA or vehicle infusions into the BLC. Subsequently, they were tested repeatedly for extinction of cocaine-CPP. Post-training BLC lesions retarded extinction of cocaine-seeking behavior and cocaine-CPP, whereas pre-training lesions disrupted acquisition of cocaine-CPP. These effects did not appear to be related to changes in general activity. We suggest that pre-training BLC lesions disrupted acquisition of cocaine-CPP by impairing assignment of incentive value to cocaine-paired stimuli, whereas post-training BLC lesions disrupted extinction of cocaine-conditioned behaviors by impairing the assessment of the current incentive value of cocaine paired stimuli. PMID- 11852028 TI - Involvement of the suprachiasmatic nucleus in body temperature modulation by food deprivation in rats. AB - Recently we found that food-deprived rats kept under a light-dark cycle showed a progressive reduction in body temperature during the light phase on each subsequent day while body temperature in the dark phase did not differ from baseline values. In this study, we investigated the effect of lesioning the hypothalamic suprachiasmatic nucleus (SCN) on body temperature modulation by food deprivation. In the SCN-lesioned rats in which daily rhythms of body temperature and activity were abolished, body temperature was unchanged by food deprivation. We also examined the effect of food deprivation on the daily changes in Fos expression in the SCN. Under normal fed conditions the number of SCN cells expressing Fos is high during the day and low at night. Food deprivation attenuated the amplitude of this daily change in Fos expression in the SCN. This tendency was prominent in the dorsal part of the SCN, while the ventral part showed no effect of food deprivation. These findings suggest that the SCN plays some role in body temperature modulation due to food deprivation. PMID- 11852029 TI - Expression of connexin 45 in the olfactory system. AB - Gap junctions represent an important mode of intercellular communication. Connexin 45 (Cx45) is a member of the connexin family that forms gap junctions between adjacent cells. In this study, we demonstrate the expression of Cx45 in the olfactory epithelium and olfactory bulb in adult mice. Reverse transcription polymerase chain reaction amplification of total RNA from mouse turbinates and olfactory bulb yielded cDNA fragments partially encoding for Cx45. In situ hybridization using Cx45 cRNA probes revealed that hybridization products were more abundant in the olfactory epithelial layer than in the lamina propria underneath the epithelium. In the olfactory epithelial layer, hybridization signals were relatively intense in a band spreading from the basal cell layer to 4/5 of the distance from the basal cell layer to the apical process. The distribution of cells positive for Cx45 mRNA is largely overlapping with that of cells expressing olfactory marker protein mRNA, indicating that a substantial number of mature olfactory neurons express Cx45 mRNA. In the olfactory bulb, cells with large nuclei in the mitral cell layer, presumably mitral cells, express Cx45 mRNA. Immunoblotting with an antibody recognizing Cx45 revealed a band at approximately 46 kDa in homogenates of mouse turbinates and olfactory bulb. Immunohistochemical studies showed fine immunoreactive puncta in the olfactory epithelium. Immunoreactivity was observed surrounding cell bodies and the proximal processes of mitral cells in the olfactory bulb. The data suggest that Cx45 is a neuronal connexin that is expressed in mature neurons in adult mice. Our study implicates a functional role for Cx45 in the olfactory system deserving future study. PMID- 11852030 TI - Spontaneous and odour evoked activity in single avian olfactory bulb neurones. AB - Extracellular single unit recordings were made from various depths in the left olfactory bulb of ten anaesthetised, freely breathing adult hens (Gallus domesticus) using glass insulated tungsten microelectrodes. The 66 spontaneously active neurons recorded had widely variable firing rates (mean 4.9 spikes/s, range 0.1-32.4 spikes/s) and variable temporal firing patterns (regular, bursting and random discharge). Interspike interval histograms were constructed for each unit and tested for goodness of fit to theoretical distributions. The activity of 23 units fitted gamma distributions, six units fitted lognormal distributions, three units fitted Weibull distributions, one unit fitted an exponential distribution and 11 units had bimodal distributions. Responses of some units to odour stimuli (Clove oil, Geraniol, Limonene and Ammonia all at 10% vapour saturation) delivered directly to the olfactory epithelium were examined. Odour stimulation modified spontaneous activity in 29 of the 44 units tested, causing either inhibition (56%) or excitation (44%) of firing. Of the 35 units stimulated with more than one odour, 15 responded to multiple stimuli, showing excitation or inhibition only or both inhibition and excitation, depending on the odour applied. Ammonia most commonly elicited a response (70% of applications) and all the odours were capable of inducing both inhibition and excitation of spontaneous firing. Examination of transverse sections of adult hen olfactory bulb in the region corresponding to the recording sites allowed unit recording depth to be related to cellular layers. There was some evidence of different neurone properties in each bulb layer. The spontaneous activity and odour responses observed resemble those seen in other vertebrate groups and the firing rates observed are between those previously reported for mammals and reptiles. This study provides detailed physiological evidence for a well-developed functional olfactory system in an avian species. PMID- 11852031 TI - Reduced serotonin release and serotonin uptake sites in the rat nucleus accumbens and striatum after prenatal cocaine exposure. AB - This study was designed to assess the effects of prenatal cocaine exposure on the development of the serotonergic system. Pregnant Sprague-Dawley rats received daily sc injections of either cocaine (30 mg/kg) or saline from gestation day 7 (GD 7) to GD 20. At 1 week postnatal, all pups were killed and tissues containing the striatum and nucleus accumbens dissected out. In superfusion experiments, tissue slices were incubated with [3H]serotonin ([3H]5-HT) for 30 min and then superfused. The [3H]5-HT release was induced by exposures to the following treatments: electrical stimulations (20 mA or 40 mA, 0.5 Hz, 4 min), the medium containing 15 or 30 mM potassium (2 min), fenfluramine (1 or 2 microM for 2 min), para-chloroamphetamine (1 or 2 microM for 2 min), methiothepin (1 or 2 microM for 2 min), and fluoxetine (1 or 2 microM for 2 min). The results showed that the treatment-induced [3H]5-HT releases were all significantly less pronounced in the pups prenatally exposed to cocaine than in those prenatally exposed to saline regardless of the mechanisms by which the treatment increases extracellular 5-HT. Saturation analysis showed that the Bmax of [3H]citalopram binding sites was also significantly lower in the pups prenatally treated with cocaine than in those prenatally treated with saline. The results are consistent with the concept that less serotonergic innervation may exist in the examined brain areas of cocaine treated offspring at 1 week postnatal, and support the hypothesis that prenatal cocaine exposure affects the postnatal development of the serotonergic system. PMID- 11852032 TI - Effects of intracerebroventricular injection of glucagon like peptide-1 and its related peptides on serotonin metabolism and on levels of amino acids in the rat hypothalamus. AB - High concentrations of glucagon-like peptide-1 (7-36) amide (GLP-1) and its specific receptor (GLP-1R) have been found in the rat hypothalamus. In this study the actions of GLP-1 and its related peptides, exendin-4 (GLP-1R agonist), exendin (9-39) (GLP-1R antagonist) and GLP-1 (9-36) amide (the major GLP-1 metabolite) on levels of serotonin (5-HT), 5-hydroxyindolacetic acid (5-HIAA) and amino acids (Glu, Asp, Gln, Gly, Tyr, Trp, GABA) in the hypothalamus were investigated. Intracerebroventricular (ICV) injection of GLP-1 (4 nmol) produced a significant reduction in levels of 5-HT (54%) and all measured amino acids (34 to 56%) compared with saline injected controls, whereas exendin (9-39) (4 nmol) was ineffective. ICV injection of exendin-4 produced a significant reduction in the levels of 5-HT, 5-HIAA, Trp, Glu, and Tyr. ICV injection of GLP-1(9-36) amide showed a statistically significant increase in the level of 5-HT, 5-HIAA and all the amino acids tested in this study. Prior administration of exendin (9-39) or GLP-1 (9-36) amide blocked the effects of GLP-1 on the levels of 5-HT and the amino acids. These data are consistent with exendin-4 being a GLP-1R agonist and exendin (9-39) being a specific GLP-1R antagonist. GLP-1 (9-36) amide, a primary metabolite of GLP-1, appears to act as an endogenous antagonist at the GLP-1R. PMID- 11852034 TI - Protein kinase C activation is required for the lead-induced inhibition of proliferation and differentiation of cultured oligodendroglial progenitor cells. AB - Lead (Pb) is a common neurotoxicant of major public health concern. Previous studies revealed that cultured oligodendrocyte progenitor cells (OPCs) are highly vulnerable to Pb toxicity. The present study examines the effect of Pb on the survival, proliferation and differentiation of OPCs in vitro. Dose-response studies showed that> or = l5-10 microM Pb is cytotoxic to OPCs within 24 h. However, 1 microM of Pb was found to inhibit the proliferation and differentiation of OPCs without affecting cell viability. Pb markedly decreased the proliferative capability of OPCs and inhibited cell-intrinsic lineage progression of OPCs at a late progenitor stage. The Pb-induced decrease of proliferation and differentiation was abolished by inhibition of protein kinase C (PKC) with bisindolylmaleimide I, while the effect of the PKC-activating agent phorbol-12,13-didecanoate was potentiated by Pb. Furthermore, Pb exposure of OPCs caused the translocation of PKC from the cytoplasm to membrane without an increase in total cellular PKC enzymic activity. These results indicate that Pb inhibits the proliferation and differentiation of oligodendrocyte lineage cells in vitro through a mechanism requiring PKC activation. PMID- 11852033 TI - Plasticity of GABA(a) system during ageing: focus on vestibular compensation and possible pharmacological intervention. AB - The lesion of the vestibular end organ evokes static and dynamic symptoms, which spontaneously regress during a complex process known as 'vestibular compensation'. Vestibular compensation is age-dependent and involves several transmitter-identified pathways in the central nervous system. In this paper we studied the time course of vestibular compensation in adult (3 months) and old (24 months) rats and correlated behavioral recovery with modifications of glutamic acid decarboxylase (GAD) mRNA expression and benzodiazepine receptor density in different brain areas. Compensation in adult rats was complete 28 days after hemilabyrinthectomy, whereas old rats still showed significant behavioral impairment. A higher GABAergic tone was found in old rats, as indicated by higher benzodiazepine receptor density in lateral vestibular nucleus and higher mRNA level for glutamic acid decarboxylase in cerebral cortex and medial vestibular nucleus. In adult, compensated rats, benzodiazepine receptor density in the vestibular nuclei was normal 28 days after lesion, whereas GAD mRNA level was higher in anterior cingulate cortex, only. On the contrary, these parameters were still altered in anterior cingulate and somatosensory cortex, basal ganglia, vestibular nuclei and cerebellum in old rats 28 days after vestibular lesion. We also evaluated the effect of the ergoline derivative nicergoline on behavioral and neurochemical correlates of vestibular compensation in old rats. Nicergoline treatment attenuated the severity of oculomotor and postural symptoms after vestibular lesion and reversed most of these age- and lesion-induced alterations in GAD mRNA expression. Thus, lesion-related alterations of the GABAergic transmission and behavioral profile after vestibular lesion are age-dependent. PMID- 11852035 TI - Analysis of opioid receptor subtype antagonist effects upon mu opioid agonist induced feeding elicited from the ventral tegmental area of rats. AB - The present study examined opioid receptor(s) mediation of feeding elicited by mu opioid agonists in the ventral tegmental area using general or selective opioid antagonist pretreatment. Naltrexone as well as equimolar doses of selective mu and kappa, but not delta opioid antagonists in the ventral tegmental area significantly reduced mu agonist-induced feeding, indicating a pivotal role for these receptor subtypes in the full expression of this response. PMID- 11852036 TI - Projection from the perirhinal cortex to the frontal motor cortex in the rat. AB - Stimulation of the anterior perirhinal cortex (PERa) induced marked surface negative and depth-positive field potentials in the rat frontal motor cortex (MC) including the rostral and caudal forelimb areas. Injection of biotinylated dextran into the PERa densely labeled axon terminals in the superficial layers of the MC, where vigorous unit responses were evoked after PERa stimulation, indicated that the perirhinal-frontal projection preferentially activates the superficial layer neurons of the MC. PMID- 11852037 TI - Upregulation in astrocytic connexin 43 gap junction levels may exacerbate generalized seizures in mesial temporal lobe epilepsy. AB - Following brain injury, and during the process of neurodegeneration, a reactive astrocytic proliferation occurs. This is accompanied by an increase in the synthesis of neuropeptides, cytokines, growth factors and glial fibrillary acidic protein (GFAP), a cell-specific marker for reactive astrocytes. Astrocytes are extensively coupled by gap junctions of the Cx43 connexin subtype. Several studies have shown that in severe trauma, coupling between astrocytes may add to the spread of the damaged area. In this study we ask whether the astrocytosis which is a feature of other neurodegenerative diseases also occurs in mesial temporal lobe epilepsy (MTLE) and whether it is accompanied by an increase in astrocytic communication through an upregulation of Cx43 gap junction channel proteins. In order to examine the astrocytic response and the expression pattern of Cx43 protein, double immunohistochemical labeling studies were undertaken using antibodies against GFAP and Cx43 applied to human hippocampal tissue resected from patients with MTLE, and to normal human control hippocampal tissue. Immunofluorescent labeling of astrocytes and Cx43 was examined using confocal laser scanning microscopy. The images obtained were quantitatively analysed and reconstructed using three-dimensional volume rendering. The results of this study have established that not only is astrocytosis greater in MTLE-affected tissues than previously suggested, but it is accompanied by a highly significant increase in astrocytic Cx43 protein levels. We hypothesize that this surprisingly large upregulation in Cx43 may exacerbate generalized seizures in the progression of MTLE. PMID- 11852038 TI - Pointing movements may be produced in different frames of reference depending on the task demand. AB - Movements are likely guided by the nervous system in task-specific spatial frames of reference (FRs). We tested this hypothesis by analyzing fast arm pointing movements involving the trunk made to targets located within the reach of the arm. In the first experiment, subjects pointed to a motionless target and, in the second experiment, to a target moving synchronously with the trunk. Vision of the arm and targets was prevented before movement onset. Each experiment started after three to five training trials. In randomly selected trials of both experiments, an electromagnet device unexpectedly prevented the trunk motion. When the trunk was arrested, the hand trajectory and velocity profile remained invariant in an FR associated with the experimental room in the first or in an FR moving with the trunk in the second experiment. Substantial changes in the arm interjoint coordination in response to the trunk arrest were observed in the first but not in the second experiment. The results demonstrate the ability of the nervous system to rapidly adapt behavior at the joint level to transform motor performance from a spatial FR associated with the environment to one associated with the body. A theoretical framework is suggested in which FRs are considered as pre-existing neurophysiological structures permitting switching between different FRs and guiding multiple joints and muscles without redundancy problems. PMID- 11852039 TI - Immunohistochemical study of p53-associated proteins in rat brain following lithium-pilocarpine status epilepticus. AB - Activation of the p53-stress response pathway has been implicated in excitotoxic neuronal cell death. Recent studies have demonstrated an age-dependent induction of both p53 mRNA and protein in the rat brain following lithium-pilocarpine mediated status epilepticus (LPSE). We investigated whether other proteins that have been shown to participate in the p53 cascade are induced by LPSE. We used immunohistochemistry to examine the expression of Mdm2, Bax, CD95/Fas/APO-1, ATM, Ref-1 and ubiquitin. A significant increase in nuclear Mdm2 immunoreactivity, which colocalized with p53, was observed in cells within hippocampal pyramidal cell layers, dentate gyrus, piriform cortex, amygdala and thalamus. Dual immunofluorescence microscopy revealed a reduction in free ubiquitin expression in cells with p53 and Mdm2 accumulation. Increased immunoreactivity for CD95/Fas/APO-1 and Bax was also detected in the same p53-positive cells. Moreover, expression of Ref-1 and ATM, which are involved in the response to oxidative stress-induced DNA damage and regulation of p53 function, were increased. Colocalization of Ref-1 and p53 suggests that Ref-1 might activate p53 function in LPSE-induced neurodegeneration. In contrast, ATM immunoreactivity was predominantly cytoplasmic suggesting that ATM may not directly modulate p53 activity in injured neurons. These results extend our previous observations with regard to activation and stabilization of p53 in injured central nervous system neurons. The data indicate that p53 induction following LPSE may activate downstream pro-apoptotic genes leading to neurodegeneration. PMID- 11852040 TI - A mouse-like retinal cone phenotype in the Syrian hamster: S opsin coexpressed with M opsin in a common cone photoreceptor. AB - Previous immunocytochemical, physiological, and molecular studies have reported that the Syrian hamster lacks a shortwave-sensitive (S) cone photopigment but retains circadian responses to ultraviolet (UV) light. Using opsin antibodies and a sensitive detection protocol, we here show that S opsin immunoreactivity colocalizes with M opsin immunoreactivity in a common type of cone photoreceptor. S opsin signal within individual cone outer segments is low and continuously decreases from the ventral to dorsal retina. Only double-labeled cones were found. During development, S opsin expression precedes that of M opsin, but there is no indication of transdifferentiation. Our results imply that in the Syrian hamster low levels of S opsin colocalize with M opsin in a common cone phenotype. We suggest that, similar to other murid rodents, the S pigment absorbs maximally in the UV range, and thus may contribute to mediating the circadian response of the Syrian hamster to UV light. PMID- 11852041 TI - Regulated and unregulated mitochondrial permeability transition pores: a new paradigm of pore structure and function? AB - Cyclosporin A (CsA) inhibits the mitochondrial permeability transition (MPT), but not always. To characterize the CsA-sensitive and -insensitive MPT, rat liver mitochondria were exposed to low and high doses of various MPT inducers. Mitochondrial swelling, cyclophilin D membrane binding and permeability transition (PT) pore diameter were measured. The results indicate two conductance modes for the PT pore: one activated by Ca(2+) and inhibited by CsA and Mg(2+) and the other unregulated. We propose a new model of pore formation and gating in which PT pores form by aggregation of misfolded integral membrane proteins damaged by oxidant and other stresses. Chaperone-like proteins initially block conductance through these misfolded protein clusters; however, increased Ca(2+) opens these regulated PT pores, an effect blocked by CsA. When protein clusters exceed chaperones available to block conductance, unregulated pore opening occurs. PMID- 11852042 TI - Shigella apyrase--a novel variant of bacterial acid phosphatases? AB - A virulence-associated ATP diphosphohydrolase activity in the periplasm of Shigella, identified as apyrase, was found to be markedly similar to bacterial non-specific acid phosphatases in primary structure. When the Shigella apyrase sequence was threaded in to the recently published 3D structure of the highly similar (73%) Escherichia blattae acid phosphatase it was found to have a highly overlapping 3D structure. Our analysis, which included assays for phosphatase, haloperoxidase and catalase activities, led us to hypothesize that Shigella apyrase might belong to a new class of pyrophosphatase originating as one more variant in the family of bacterial non-specific acid phosphatases. It revealed interesting structure-function relationships and probable roles relevant to pathogenesis. PMID- 11852043 TI - Synthesis, membrane insertion and assembly of the chloroplast-encoded D1 protein into photosystem II. AB - Rapid light-dependent turnover of the chloroplast-encoded D1 protein maintains photosystem II (PS II) functional over a wide range of light intensities. Following initiation of psbA mRNA translation, the elongating D1 is targeted, possibly by chloroplast signal recognition particle 54 (cpSRP54), to the thylakoid cpSecY translocation channel. Transmembrane domains of nascent D1 start interacting with other PS II core proteins already during the translocation process to ensure an efficient assembly of the multiprotein membrane complex. Here we review the progress recently made concerning the synthesis, targeting, membrane insertion and assembly to PS II of the chloroplast-encoded D1 protein and discuss the possible convergence of targeting and translocation of chloroplast- and nuclear-encoded thylakoid proteins. PMID- 11852044 TI - Structure and functional properties of the ubiquitin binding protein p62. AB - Several highly conserved p62 homologs have recently been isolated, e.g. the rat atypical protein kinase C-interacting protein (ZIP), the murine A170/signal transduction and adapter protein, and the human p62, a protein that binds the Src homology 2 domain of p56(lck). These proteins share striking similarity in amino acid sequence and structural motifs, thereby suggesting conserved functional properties. ZIP/p62 has been shown to play an important role as a scaffold leading to the activation of the transcription factor nuclear factor kappaB. In addition, a nuclear form of p62 has been characterized that can serve as a transcriptional co-activator. Moreover, p62 is capable of binding ubiquitin (Ub) non-covalently through its Ub-associated domain. In this review, we will focus on the structure and function of ZIP/p62. PMID- 11852045 TI - Cellular prion protein: on the road for functions. AB - Cellular prion (PrPc) is a plasma membrane glycosyphosphatidylinositol-anchored protein present in neurons but also in other cell types. Protein conservation among species suggests that PrPc may have important physiological roles. Cellular and molecular approaches have established several novel features of the regulation of PrPc expression, cellular trafficking as well as its participation in copper uptake, protection against oxidative stress, cell adhesion, differentiation, signaling and cell survival. It is therefore likely that PrPc plays pleiotropic roles in neuronal and non-neuronal cells, and as such the loss of function of PrPc may be an important component of various diseases. PMID- 11852046 TI - Catalytic site occupancy during ATP synthase catalysis. AB - An early proposal was that for rapid ATP synthesis by the rotational ATP synthase, a specific second site must bind ADP and P(i), and for rapid ATP hydrolysis a different second site must bind ATP. Such bi-site activation was considered to occur whether or not an ADP or ATP was at a third site. In contrast, a more recent proposal is that rapid ATP hydrolysis requires that all three sites have bound ADP or ATP present. However, discovery that one second site binds ADP better than ATP, together with other data and considerations support the earlier proposal. The retention or rebinding of ADP can explain why three sites fill during hydrolysis as ATP concentration is increased although bi site activation still prevails. PMID- 11852047 TI - Quinones in long-lived clk-1 mutants of Caenorhabditis elegans. AB - Ubiquinone (UQ) (coenzyme Q) is a lipophilic redox-active molecule that functions as an electron carrier in the mitochondrial electron transport chain. Electron transfer via UQ involves the formation of semiubiquinone radicals, which causes the generation of superoxide radicals upon reaction with oxygen. In the reduced form, UQ functions as a lipid-soluble antioxidant, and protects cells from lipid peroxidation. Thus, UQ is also important as a lipophilic regulator of oxidative stress. Recently, a study on long-lived clk-1 mutants of Caenorhabditis elegans demonstrated that biosynthesis of UQ is dramatically altered in mutant mitochondria. Demethoxy ubiquinone (DMQ), that accumulates in clk-1 mutants in place of UQ, may contribute to the extension of life span. Here we elucidate the possible mechanisms of life span extension in clk-1 mutants, with particular emphasis on the electrochemical property of DMQ. Recent findings on the biochemical function of CLK-1 are also discussed. PMID- 11852048 TI - Copper-induced conformational change in a marsupial prion protein repeat peptide probed using FTIR spectroscopy. AB - We report the first Fourier transform infrared analysis of prion protein (PrP) repeats and the first study of PrP repeats of marsupial origin. Large changes in the secondary structure and an increase in hydrogen bonding within the peptide groups were evident from a red shift of the amide I band by >7 cm(-1) and an approximately five-fold reduction in amide hydrogen-deuterium exchange for peptide interacting with Cu(2+) ions. Changes in the tertiary structure upon copper binding were also evident from the appearance of a new band at 1564 cm( 1), which arises from the ring vibration of histidine. The copper-induced conformational change is pH dependent, and occurs at pH >7. PMID- 11852049 TI - Point mutations confer loss of ATP-induced human P2X(7) receptor function. AB - Residues considered essential for ATP binding to the human P2X(7) receptor (hP2X(7)R) were investigated. HEK293 cells or Xenopus oocytes were transfected with wild-type or site-directed mutants of hP2X(7)R constructs and channel/pore activity measured in the presence of ATP or 2',3'-O-(4-benzoylbenzoyl)-ATP (BzATP). Barium uptake and ethidium influx into HEK293 cells were abolished in cells expressing K193A and K311A mutants, and were partially reduced in cells expressing mutant P210A. K193A and K311A mutations also completely abolished responses to ATP and BzATP in Xenopus oocytes as measured by electrophysiology. These results indicate that K193 and K311 are essential residues in ATP binding in the hP2X(7)R. PMID- 11852050 TI - The orientation of the antibiotic peptide maculatin 1.1 in DMPG and DMPC lipid bilayers. Support for a pore-forming mechanism. AB - Maculatin 1.1 is an antimicrobial peptide isolated from the Australian tree frog Litoria genimaculata that adopts an amphipathic, alpha-helical structure in solution. Its orientation and conformation when incorporated to pre-formed DMPG (1,2-dimyristoyl-sn-glycero-3-phosphoglycerol) and DMPC (1,2-dimyristoyl-sn glycero-3-phosphocholine) vesicles was determined using polarised Fourier transform infrared-attenuated total reflection infrared and deuterium exchange experiments. For DMPG membranes, our results show insertion of 70% of the maculatin 1.1 molecules, with an angle of insertion of approximately 35 degrees to the membrane normal and with a predominant alpha-helical structure. These results suggest that maculatin 1.1 acts through a pore-forming mechanism to lyse bacterial membranes. A similar degree of insertion in DMPG (65%) and alpha helical structure was observed for a biologically inactive, less amphipathic maculatin 1.1 analogue, P15A, although the helix tilt was found to be greater (46 degrees) than for maculatin 1.1. Similar experiments performed using DMPC liposomes showed poor insertion, less than 5%, for both maculatin 1.1 and its analogue. In addition, the shape of the amide I band in these samples is consistent with alpha-helix, beta-structure and disordered structures being present in similar proportion. These results clearly show that maculatin 1.1 inserts preferentially in negatively charged membranes (DMPG) which mimic the negatively charged membrane of Gram-positive bacteria. We attribute the high percentage of insertion of the biologically inactive analogue in DMPG to the fact that its concentration on the membrane surface in our experiments is likely to be much higher than that found in physiological conditions. PMID- 11852051 TI - Anion exchanger isoform 2 operates in parallel with Na(+)/H(+) exchanger isoform 1 during regulatory volume decrease of human cervical cancer cells. AB - Intracellular pH (pH(i)) homeostasis was investigated in human cervical cancer SiHa cells undergoing regulatory volume decrease (RVD) to determine which transport systems were involved. Using isoform-specific primers, mRNA transcripts of Na(+)/H(+) exchanger isoform 1 (NHE1) and isoform 3 were identified by reverse transcriptase polymerase chain reaction (RT-PCR) and the results confirmed by Western immunoblotting. From anion exchanger isoforms 1-3 (AE1-3), only the mRNA transcript of AE2 was identified by RT-PCR and the identity was confirmed by digestion with a specific restriction endonuclease. SiHa cells loaded with the fluorescent dye 2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein and resuspended in isotonic media showed a stable pH(i). In contrast, a gradual internal acidification took place following resuspension in hypotonic media. The NHE inhibitors, HOE694 (10 microM) and amiloride (1 mM), showed a similar potency in enhancing the rate and extent of the hypotonicity-induced internal acidification. The absence of extracellular Na(+) also substantially enhanced the acidification during RVD. These results suggest that internal acidification during RVD is mainly compensated by the operation of NHE1. Extracellular Cl(-) was critically necessary for the pH(i) acidification during RVD. The hypotonicity-induced acidification was significantly attenuated by 100 microM 4,4' diisothiocyanostilbene-2,2'-disulfonic acid, a concentration inhibiting more than 90% AE activity. This indicates that AE2 mediates a net Cl(-) influx with compensating HCO(3)(-) efflux during RVD. We conclude that AE2 operates in parallel with NHE1 to regulate pH(i) during RVD of human cervical cancer cells. PMID- 11852052 TI - Inhibitory actions of the selective serotonin re-uptake inhibitor citalopram on HERG and ventricular L-type calcium currents. AB - Using whole-cell patch clamp recording of heterologous HERG-mediated currents in transfected mammalian cells, we observed that the selective serotonin re-uptake inhibitor citalopram blocks HERG with an IC(50) of 3.97 microM. This is slightly less potent than fluoxetine in our system (IC(50) of 1.50 microM). In isolated guinea pig ventricular cardiomyocytes citalopram inhibited L-type calcium current (I(Ca,L)). The voltage dependence of I(Ca,L) inactivation in the presence of 100 microM citalopram was shifted significantly leftward. As a result, the I(Ca,L) 'window' in citalopram was found to be (a) smaller and (b) leftward-shifted compared to control. The effects of citalopram on both calcium current amplitude and the I(Ca,L) 'window' may help to explain citalopram's good cardiac safety profile, given its propensity to block HERG at excessive dosages. PMID- 11852053 TI - Identification of functional type 1 ryanodine receptors in mouse dendritic cells. AB - Ca(2+) signaling plays an important role in the function of dendritic cells (DC), the specialized antigen-presenting cells of the immune system. Here we describe functional ryanodine receptor (RyR) Ca(2+) release channels in murine, bone marrow-derived DC. RT-PCR analysis identified selective expression of the type 1 RyR, with higher levels detected in immature rather than mature DC. The RyR activators caffeine, FK506, ryanodine and 4-chloro-m-cresol mobilized Ca(2+) in DC, and responses to 4-chloro-m-cresol were inhibited by dantrolene. Furthermore, activation of RyRs both inhibited subsequent inositol trisphosphate-mediated Ca(2+) release and provoked store-operated Ca(2+) entry, suggesting a functional interaction between these intracellular Ca(2+) channels. Thus, the RyR1 channel may play an intrinsic role in Ca(2+) signaling in DC. PMID- 11852054 TI - 2-Oxoglutarate increases the binding affinity of the NtcA (nitrogen control) transcription factor for the Synechococcus glnA promoter. AB - The cyanobacterial NtcA global nitrogen regulator belongs to the catabolite activator protein (CAP) family and activates transcription of nitrogen assimilation genes in response to nitrogen step-down. The binding affinity of NtcA towards a DNA fragment carrying the promoter of the glnA gene from Synechococcus sp. PCC 7942, analyzed in vitro by band-shift assay, was increased five-fold by 2-oxoglutarate in the presence of Mg(2+) ions. The 2-oxoglutarate effect peaked at about 0.6 mM, a rather physiological concentration for this compound under nitrogen-limiting conditions, and could be partially reproduced by 3-oxoglutarate but not by oxaloacetate or glutamate. These results suggest 2 oxoglutarate as a signal of the C to N balance of the cells to regulate NtcA activity and provide a new example of regulation in the versatile CAP family of proteins. PMID- 11852055 TI - NF-kappaB and ERK cooperate to stimulate DNA synthesis by inducing ornithine decarboxylase and nitric oxide synthase in cardiomyocytes treated with TNF and LPS. AB - We previously reported that tumor necrosis factor-alpha (TNF) and lipopolysaccharide (LPS) stimulate DNA synthesis in chick embryo cardiomyocytes (CM) via nitric oxide and polyamine biosynthesis. Here we show an involvement of nuclear factor-kappaB (NF-kappaB) in the induction of nitric oxide synthase (NOS) and ornithine decarboxylase (ODC), the key enzyme in polyamine biosynthesis. In addition NF-kappaB activation appears to favor survival of CM by reducing caspase activation. TNF and LPS also stimulate phosphorylation of extracellular signal regulated kinase (ERK), which is required for the changes in ODC and caspase activity, but not for NOS induction or NF-kappaB activation. In conclusion, these results indicate that NF-kappaB, in cooperation with ERK, plays a pivotal role in the growth stimulating effects of TNF and LPS, leading to the induction of both ODC and NOS and to the reduction of caspase activity. PMID- 11852056 TI - The proton-pumping NADH:ubiquinone oxidoreductase (complex I) of Aquifex aeolicus. AB - The proton-pumping NADH:ubiquinone oxidoreductase, also called complex I, is the first energy-transducing complex of many respiratory chains. Homologues of complex I are present in the three domains of life. Here, we report the properties of complex I in membranes of the hyperthermophilic bacterium Aquifex aeolicus. The complex reacted with NADH but not with NADPH and F(420)H(2) as electron donors. Short-chain analogues of ubiquinone like decyl-ubiquinone and ubiquinone-2 were suitable electron acceptors. The affinities towards NADH and ubiquinone-2 were comparable to the ones obtained with the Escherichia coli complex I. The reaction was inhibited by piericidin A at the same concentration as in E. coli. The complex showed an unusual pH optimum at pH 9 and a maximal rate at 80 degrees C. We found no evidence for the presence of an alternative, single subunit NADH dehydrogenase in A. aeolicus membranes. The NADH:ferricyanide reductase activity of detergent extracts of A. aeolicus membranes sedimented as a protein with a molecular mass of approximately 550 kDa. From the data we concluded that A. aeolicus contains a NADH:ubiquinone oxidoreductase resembling complex I of mesophilic bacteria. PMID- 11852057 TI - Peroxisome proliferator-activated receptor (PPAR) agonists decrease lipoprotein lipase secretion and glycated LDL uptake by human macrophages. AB - Lipoprotein lipase (LPL) acts independently of its function as triglyceride hydrolase by stimulating macrophage binding and uptake of native, oxidized and glycated LDL. Peroxisome proliferator-activated receptors (PPARs) are nuclear receptors expressed in monocyte/macrophages, where they control cholesterol homeostasis. Here we study the role of PPARs in the regulation of LPL expression and activity in human monocytes and macrophages. Incubation of human monocytes or macrophages with PPARalpha or PPARgamma ligands increases LPL mRNA and intracellular protein levels. By contrast, PPAR activators decrease secreted LPL mass and enzyme activity in differentiated macrophages. These actions of PPAR activators are associated with a reduced uptake of glycated LDL and could influence atherosclerosis development associated with diabetes. PMID- 11852058 TI - Serine (threonine) phosphatase(s) acting on cAMP-dependent phosphoproteins in mammalian mitochondria. AB - Immunochemical and functional evidence showing the existence in the inner membrane and matrix fraction of mammalian mitochondria of serine/threonine phosphatases acting on cAMP-dependent phosphoproteins is presented. Mg(2+) dependent Ca(2+)-inhibitable PP2C phosphatase, associated to the inner membrane, dephosphorylates the 18 kDa (NDUFS4 gene) of complex I. PMID- 11852059 TI - Involvement of the N-terminus of Bax in its intracellular localization and function. AB - We have identified, using site-directed mutagenesis, a proline located at position 13 of Baxalpha (Bax) as crucial for the maintenance of its cytosolic conformation. The substitution of this proline by a valine results in a strong binding of Bax to mitochondria and to conformational changes monitored by a decreased sensitivity of Bax to mild proteolysis and the enhancement of its oligomerization state. Deletion of the C-terminus of Bax does not modify its intracellular localization. On the other hand, the pro-apoptotic activity of Bax is enhanced by a deletion of the C-terminus in the absence of the N-terminus but is decreased in its presence. These results suggest that both extremities functionally interact to control the activity but not the subcellular localization of Bax. PMID- 11852060 TI - Role of glycosylation in hyperphosphorylation of tau in Alzheimer's disease. AB - In Alzheimer's disease (AD) brain, microtubule-associated protein tau is abnormally modified by hyperphosphorylation and glycosylation, and is aggregated as neurofibrillary tangles of paired helical filaments. To investigate the role of tau glycosylation in neurofibrillary pathology, we isolated various pools of tau protein from AD brain which represent different stages of tau pathology. We found that the non-hyperphosphorylated tau from AD brain but not normal brain tau was glycosylated. Monosaccharide composition analyses and specific lectin blots suggested that the tau in AD brain was glycosylated mainly through N-linkage. In vitro phosphorylation indicated that the glycosylated tau was a better substrate for cAMP-dependent protein kinase than the deglycosylated tau. These results suggest that the glycosylation of tau is an early abnormality that can facilitate the subsequent abnormal hyperphosphorylation of tau in AD brain. PMID- 11852061 TI - Identification of functional VEGF receptors on human platelets. AB - Platelets secrete platelet-derived growth factor (PDGF) and vascular endothelial growth factor (VEGF) upon stimulation. We have demonstrated that platelets have functionally active PDGF alpha-receptors, a transmembrane tyrosine kinase involved in negative feedback regulation. Here we demonstrate the presence of the related VEGF receptors fms-like tyrosine kinase-1 and kinase-insert domain region on human platelets. VEGF itself did not cause platelet aggregation. However, addition of exogenous VEGF to SFRLLN or thrombin-stimulated platelets potentiated platelet aggregation. Moreover, thrombin-induced phosphoinositide 3-kinase and mitogen-activated protein kinase activity were enhanced in the presence of VEGF. PMID- 11852062 TI - Characterization of a monomeric disintegrin, ocellatusin, present in the venom of the Nigerian carpet viper, Echis ocellatus. AB - Ocellatusin is a new RGD-containing short monomeric disintegrin. It is a better inhibitor of alpha(5)beta(1) integrin and a more potent inducer of the expression of a ligand-induced binding site epitope on beta(1) integrin subunit than echistatin. In further contrast to echistatin, ocellatusin has a direct chemotactic stimulus on human neutrophils in vitro. The distinct effects of these two close evolutionarily related disintegrins might be explained by the presence of methionine-22 and histidine-29 in the RGD loop of ocellatusin, which are arginine and aspartic acid, respectively, in echistatin. These mutations may modulate the conformation and/or recognition properties of the integrin-binding loop of ocellatusin. PMID- 11852063 TI - The first room-temperature X-ray absorption spectra of higher oxidation states of the tetra-manganese complex of photosystem II. AB - The manganese (Mn) complex of photosystem II catalyzes water oxidation. For the first time, its advancement through the reaction cycle was monitored by time resolved X-ray absorption measurements at the Mn K-edge at room temperature. The complex was stepped through its four oxidation states by nano-second-laser flashes applied to samples exposed to the X-ray beam. Time courses of the X-ray fluorescence intensity were recorded during a flash sequence. Extended X-ray absorption fine-structure spectra were recorded with the S(1), S(2), and S(3) oxidation states highly populated. The room temperature data is compatible with the formation of a third di-mu-oxo bridge between two Mn atoms upon the S(2)- >S(3) transition. PMID- 11852064 TI - A novel approach to study the activity and stoichiometry simultaneously for microsomal pentoxyresorufin-O-dealkylase reaction. AB - A simple approach to study the activity and stoichiometry of cytochrome P-450 IIB1-catalyzed metabolism of pentoxyresorufin (PRF) has been investigated. It involves the continuous spectral analysis of reaction mixture containing PRF, microsomes from phenobarbital-induced rats and NADPH. The kinetics of NADPH consumption, PRF utilization, NADP and resorufin formation was monitored at lambda(max) of 338, 484, 260 and 572 nm, respectively. The stoichiometry of the enzyme reaction tabulated either by specific activity or by V(max) value showed that 10 molecules of NADPH were required for the conversion of one molecule of PRF to one molecule of resorufin along with 10 molecules of NADP. Further, it was observed that almost six molecules of NADPH are consumed in the incubation mixture devoid of PRF indicating the possibility of metabolism of endogenous substrates. Interestingly, the stoichiometry ratio of 1:1 for PRF and resorufin was established even in the presence of P-450 inhibitors with a lower rate of metabolism. However, the ratio of NADPH to PRF was altered in the presence of inhibitors, suggesting that the simultaneous monitoring of the substrate, electron donor and the products could be useful in understanding the modifications of stoichiometry of electron donor and substrate/product. PMID- 11852065 TI - Fractionation of cytochromes of phototrophically grown Chloroflexus aurantiacus. Is there a cytochrome bc complex among them? AB - The cytochrome-containing membrane complexes of the phototrophically grown green non-sulfur bacterium Chloroflexus aurantiacus were fractionated by anion exchange chromatography. Three cytochrome b and four cytochrome c peaks were observed. None of the separated complexes met the features of the cytochrome bc complex. Two main cytochrome b-containing complexes were further purified: a dimer of identical subunits with unknown function and a succinate:quinone oxidoreductase containing three subunit species. Two novel multisubunit complexes, similar to each other, with two heme c-bearing subunits were also purified. PMID- 11852066 TI - Light control over the size of an antenna unit building block as an efficient strategy for light harvesting in photosynthesis. AB - It was shown that an increase in the bacteriochlorophyll (BChl) c antenna size observed upon lowering growth light intensities led to enhancement of the hyperchromism of the BChl c Q(y) absorption band of the green photosynthetic bacterium Chloroflexus aurantiacus. With femtosecond difference absorption spectroscopy, it was shown that the amplitude of bleaching of the oligomeric BChl c Q(y) band (as compared to that for monomeric BChl a) increased with increasing BChl c content in chlorosomes. This BChl c bleaching amplitude was about doubled as the chlorosomal antenna size was about trebled. Both sets of findings clearly show that a unit BChl c aggregate in the chlorosomal antenna is variable in size and governed by the grow light intensity, thus ensuring the high efficiency of energy transfer within the BChl c antenna regardless of its size. PMID- 11852067 TI - Evidence for diversity of substrate specificity among members of the chymase family of serine proteases. AB - The term chymase is used to signify a chymotrypsin-like protease stored within the secretory granules of mast cells. Primarily based on amino acid sequence homology, 18 chymases have been identified among different animals. This study, which compares the structure of the primary specificity pocket (S1 subsite), defines a subgroup of four chymases likely to have a substrate specificity with more elastase- than chymotrypsin-like qualities. This difference is due, primarily, to finding a Val instead of a Gly at residue 199, a position corresponding to Gly216 in bovine chymotrypsin and Val216 in neutrophil and porcine elastases. Chymases with Val at 199 are found only in animals expressing multiple chymases, consistent with the premise that their substrate specificity differs from that of chymases with Gly at 199. PMID- 11852068 TI - On the pro-oxidant effects of haemozoin. AB - Haemozoin (Hz) is a haem aggregate produced in some blood-feeding organisms. There is a general belief that Hz formation would be a protective mechanism against haem toxicity. Here we show that when aggregated into Hz, haem is less deleterious than its free form. When haem was added to phosphatidylcholine (PC) liposomes, there was an intense stimulation of oxygen consumption, which did not occur when Hz was incubated with the same preparation. Evaluation of oxygen radical attack to lipids, by measurement of thiobarbituric acid reactive substances (TBARS), showed significantly lower levels of lipid peroxidation in samples containing PC liposomes incubated with Hz than with haem. However, TBARS production induced by Hz was much higher when using 2-deoxyribose (2-DR) as substrate, than with PC liposomes. Spin-trapping analysis by electron paramagnetic resonance (EPR) of Hz and tert-butylhydroperoxide (tert-BuOOH) showed that production of methoxyl and tert-butoxyl radicals was only slightly reduced compared to what was observed with haem. Interestingly, when large Hz crystals were used in 2-DR TBARS assays and tert-BuOOH EPR experiments, the pro oxidant effects of Hz were strongly reduced. Moreover, increasing concentrations of Hz did not induce erythrocyte lysis, as occurred with haem. Thus, the reduced capacity of Hz to impose radical damage seems to result from steric hindrance of substrates to access the aggregated haem, that becomes less available to participate in redox reactions. PMID- 11852069 TI - Reversible inhibition of photophosphorylation in chloroplasts by nitric oxide. AB - Nitric oxide (NO) is a bioactive molecule involved in diverse physiological functions in plants. Here we demonstrate that NO is capable of regulating the activity of photophosphorylation in chloroplasts. The electron transport activity in photosystem II determined from chlorophyll a fluorescence was inhibited by NO. NO also inhibited light-induced DeltapH formation across the thylakoid membrane. High concentrations of nitrite and nitrate did not show such inhibitory effects, suggesting that the inhibition is not due to uncoupling effects of the oxidized products of NO. ATP synthesis activity upon illumination was severely inhibited by NO (IC(50)=0.7 microM). The inhibition was found to be temporary and the activity was completely recovered by removing NO. Bovine hemoglobin and bicarbonate were effective in preventing NO-dependent inhibition of photophosphorylation. These results indicate that NO is a reversible inhibitor of photosynthetic ATP synthesis. PMID- 11852070 TI - 8-Chloro-dGTP, a hypochlorous acid-modified nucleotide, is hydrolyzed by hMTH1, the human MutT homolog. AB - The human mutT homolog, hMTH1, suppresses spontaneous mutations by degrading the endogeneous mutagen, 8-hydroxy-dGTP. We previously reported the broad substrate specificity of hMTH1, which also degrades the oxidatively damaged purine nucleotides, 2-hydroxy-dATP, 8-hydroxy-dATP, 2-hydroxy-ATP, and 8-hydroxy-GTP, in addition to 8-hydroxy-dGTP. In this paper, we describe the hMTH1 activity for 8 chloro-dGTP, which could be formed in inflamed tissue by the reaction of dGTP with hypochlorous acid, a product of myeloperoxidase from activated human neutrophils. The hMTH1 protein was mixed with 1-20 microM of 8-chloro-dGTP and 8 hydroxy-dGTP, and the reaction products were quantified by anion-exchange HPLC to measure the pyrophosphatase reaction rate. The kinetic parameters revealed that 8 chloro-dGTP was degraded by hMTH1 with 50% efficiency as compared with that of 8 hydroxy-dGTP. This result suggests that 8-chloro-dGTP is an intrinsic substrate for hMTH1. PMID- 11852071 TI - First patch, then catch: measuring the activity and the mRNA transcripts of a proton pump in individual Lilium pollen protoplasts. AB - Combining the patch-clamp method with single-cell reverse transcription polymerase chain reaction (scRT-PCR) a fusicoccin-induced current reflecting the activity of the plasma membrane H(+) ATPase of lily pollen protoplasts was measured and subsequently, the ATPase-encoding mRNAs were collected and amplified. Southern blot signals were observed in all 'patch-catch' experiments and could be detected even in 2560-fold dilutions of the pollen contents. H(+) ATPase mRNAs were detectable only in the vegetative but not in the generative cell of pollen as confirmed by immunolocalisation. In 15% of the scRT-PCR experiments, a random non-reproducibility of the PCR was observed, probably caused by varying amounts of ATPase mRNAs in the protoplasts. PMID- 11852072 TI - Normoxic induction of the hypoxia-inducible factor 1alpha by insulin and interleukin-1beta involves the phosphatidylinositol 3-kinase pathway. AB - Hypoxia-inducible factor 1 (HIF-1) is a heterodimeric DNA-binding complex of the subunits alpha and beta with relevance in O(2) and energy homeostasis. The labile component, HIF-1alpha, is not only activated by hypoxia but also by peptides such as insulin and interleukin-1 (IL-1) in normoxia. We investigated whether inhibitors of mitogen-activated protein kinase kinases (MAPKKs: PD 98059, U0126) and phosphatidylinositol 3-kinase (PI3K: LY 294002) do not only lower the hypoxia induced, but also the insulin- and IL-1-induced HIF-1alpha accumulation and HIF-1 DNA-binding in human hepatoma cell cultures (line HepG2). The results show that LY 294002 suppressed HIF-1 activation in a dose-dependent manner irrespective of the stimulus. With respect to target proteins controlled by HIF-1, the production of erythropoietin was fully blocked and that of vascular endothelial growth factor reduced following inhibition of the PI3K pathway. The role of MAPKKs in this process remained in question, because PD 98059 and U0126 did not significantly reduce HIF-1alpha levels at non-toxic doses. We propose that PI3K signaling is not only important in the hypoxic induction of HIF-1 but it is also crucially involved in the response to insulin and IL-1. PMID- 11852073 TI - Murine T cells expressing high activity of prolyl endopeptidase are susceptible to activation-induced cell death. AB - Prolyl endopeptidase (PEP) is widely distributed and thought to play an important role in the degradation of peptide hormones and neuropeptides, but its biological role is totally unknown. In this study, we examined PEP activity in subpopulations of murine T cells and found that PEP activity was significantly higher in immature thymocytes than in mature thymocytes or in peripheral T cells. Stimulation of murine peripheral T cells time-dependently increased PEP activity. Although murine T cell hybridomas exhibited high PEP activity, the PEP activity was fully inhibited by treatment with PEP inhibitor. The pretreated T cells were found to be resistant to activation-induced cell death (AICD). Similar results were obtained in murine thymocytes as well as in activated peripheral T cells. PEP activity in T cell hybridomas remained unchanged during AICD. These results suggest that T cells expressing high PEP activity are susceptible to ACID. PMID- 11852074 TI - Restoration of UV sensitivity in UV-resistant HeLa cells by antisense-mediated depletion of damaged DNA-binding protein 2 (DDB2). AB - Damaged DNA-binding activity comprises two major protein components, DDB1 and DDB2, which are implicated in the repair of ultraviolet (UV) radiation-induced DNA damage. The possible role of DDB2 as a determinant of cellular sensitivity to UV was investigated. The abundance of DDB2 in UV-resistant HeLa cell lines was increased compared with that in the parental UV-sensitive cells. Stable transfection of the resistant cells with DDB2 antisense cDNA resulted in marked depletion of DDB2 protein and restored cellular sensitivity to UV-induced apoptosis. Whereas the extent of UV-induced activation of apoptosis executioners, including DNA fragmentation factor, and caspase-3 were reduced in the UV resistant cells compared with those apparent in the sensitive cells, depletion of DDB2 from the resistant cells restored the normal activation patterns for these proteins. In contrast, overexpressing DDB2 in DDB2-depleted cells with recombinant adenovirus, which carries ddb2 cDNA, markedly inhibited the extent of UV-induced activation of DNA fragmentation factor, and caspase-3. Interestingly, a mutated form of DDB2, which is defective in interacting with DDB1 and binding to UV-damaged DNA, also markedly inhibited the activation of apoptosis executioners. These results indicate that DDB2 is a modulator of UV-induced apoptosis, and that UV resistance can be overcome by inhibition of DDB2. The findings also suggest that modulation of UV-induced apoptosis by DDB2 may be independent of DNA repair. PMID- 11852075 TI - Mai1p is essential for maturation of proaminopeptidase I but not for autophagy. AB - We here identify Mai1p, a homologue of the autophagy protein Aut10p, as a novel component essential for proaminopeptidase I (proAPI) maturation under non starvation conditions. In mai1Delta cells mature vacuolar proteinases are detectable and vacuolar acidification is normal. In mai1Delta cells autophagy occurs, though at a somewhat reduced level. This is indicated by proAPI maturation during starvation and accumulation of autophagic bodies during starvation with phenylmethylsulfonyl fluoride. Homozygous diploid mai1Delta cells sporulate, but with a slightly reduced frequency. Biologically active Ha-tagged Mai1p, chromosomally expressed under its native promoter, is at least in part peripherally membrane-associated. In indirect immunofluorescence it localizes to the vacuolar membrane or structures nearby. In some cells Ha-tagged Mai1p appears concentrated at regions adjacent to the nucleus. PMID- 11852076 TI - Regulatory interactions in Arabidopsis thaliana acetohydroxyacid synthase. AB - Acetohydroxyacid synthase (AHAS; EC 4.1.3.18) contains catalytic and regulatory subunits, the latter being required for sensitivity to feedback regulation by leucine, valine and isoleucine. The regulatory subunit of Arabidopsis thaliana AHAS possesses a sequence repeat and we have suggested previously that one repeat binds leucine while the second binds valine or isoleucine, with synergy between the two sites. We have mutated four residues in each repeat, based on a model of the regulatory subunit. The data confirm that there are separate leucine and valine/isoleucine sites, and suggest a complex pathway for regulatory signal transmission to the catalytic subunit. PMID- 11852077 TI - Molecular simulations of the large conductance mechanosensitive (MscL) channel under mechanical loading. AB - The MscL channel is a mechanosensitive channel which is gated by membrane stress or tension. Here, we describe a series of simulations which apply simulated mechanical stress to a molecular model of the MscL channel using two methods - direct force application to the transmembrane segments, and anisotropic pressure coupling. In the latter simulations, pressures less than that equivalent to a bilayer tension of 12 dyn/cm did not cause the channel to open, while pressures in excess of this value resulted in the channel opening. These results are in approximate agreement with experimental findings. PMID- 11852078 TI - Heterologous expression and topography of the main intrinsic protein (MIP) from rat lens. AB - Wild type rat lens main intrinsic protein (MIP) and MIP mutated (F73I, F75L) to resemble the glycerol facilitator of Escherichia coli in the region of the NPA1 box were used to investigate the topology of MIP in the membrane of Spodoptera frugiperda (Sf21) cells using the baculovirus expression system and expression in mouse erythroid leukaemia cells (MEL C88). Differential fixation for staining was used, with paraformaldehyde for externally exposed antigenic sites, and acetone for both externally and internally exposed protein antigenic sites. Immunofluorescence using antibodies to synthetic MIP peptides showed that wild type MIP had a six transmembrane topography. The N- and C-termini were intracellular in both expression systems, and both NPA boxes were found to be extracellular. These results show that residues around the NPA1 box can influence the folding of the MIP in the membrane, and provide structural evidence for the poor water transport properties of MIP, as the NPA boxes lie outside the plane of the membrane. PMID- 11852080 TI - In vitro PKA phosphorylation-mediated human PDE4A4 activation. AB - The PDE4 catalytic machinery comprises, in part, two divalent cations in a binuclear motif. Here we report that PDE4A4 expressed in Sf9 cells exhibits a biphasic Mg(2+) dose-response (EC(50) of 0.15 and >10 mM) in catalyzing cAMP hydrolysis. In vitro phosphorylation of PDE4A4 by the PKA-catalytic subunit increases the enzyme's sensitivity to Mg(2+), leading to 4-fold increased cAMP hydrolysis without affecting its K(m). The phosphorylation also increases the potencies of (R)- and (S)-rolipram without affecting CDP-840 and SB-207499. The results support that modulating the cofactor binding affinity of PDE4 represents a mechanism for regulating its activity. PMID- 11852079 TI - pH-Dependent channel activity of heterologously-expressed main intrinsic protein (MIP) from rat lens. AB - Wild-type rat lens main intrinsic protein (MIP) was heterologously expressed in the membrane of Spodoptera frugiperda (Sf21) cells using the baculovirus expression system and in mouse erythroid leukaemia cells (MEL C88). Both MEL and Sf21 cell lines expressing wild-type MIP were investigated for the conductance of ions using a whole cell patch clamp technique. An increase in conductance was seen in both expression systems, particularly on lowering the pH to 6.3. In Sf21 cells, addition of antibodies to the NPA1 box resulted in a reduction of current flow. These results suggest that MIP has pH-dependent ion channel activity, which involves the NPA1 box domain. PMID- 11852081 TI - Single synonymous codon substitution eliminates pausing during chloramphenicol acetyl transferase synthesis on Escherichia coli ribosomes in vitro. AB - The coding sequence for chloramphenicol acetyl transferase (CAT) contains several rare codons; three of them are ATA encoding isoleucine in positions 13, 84 and 119 of the amino acid sequence. Expression of CAT on Escherichia coli ribosomes in vitro results in mostly full-length product but also distinct smaller polypeptides from less than 3 kDa to over 20 kDa. As reported earlier, the smaller polypeptides are the predominant products, if translation is initiated with fluorophore-Met-tRNA(f). All this translational pausing is eliminated when the first ATA codon is mutated to ATC, a frequently used codon for isoleucine in E. coli. Addition of large amounts of E. coli tRNA to the coupled transcription/translation reaction does not reduce the number of pause-site peptides seen in the expression of wild-type CAT. Thus we hypothesize that the mRNA structure may be an important determinant for translational pausing. PMID- 11852082 TI - Different effects of calnexin deletion in Saccharomyces cerevisiae on the secretion of two glycosylated amyloidogenic lysozymes. AB - Both glycosylated amyloidogenic lysozymes I55T/G49N and D66H/G49N were expressed in wild-type and calnexin-disrupted Saccharomyces cerevisiae. The secretion amounts of mutant I55T/G49N were almost similar in both wild-type and calnexin disrupted S. cerevisiae. In contrast, the secretion of mutant D66H/G49N greatly increased in calnexin-disrupted S. cerevisiae, while the secretion was very low in the wild-type strain. In parallel, the induction level of the molecular chaperones BiP and PDI located in the endoplasmic reticulum (ER) was investigated when these glycosylated amyloidogenic lysozymes were expressed in wild-type and calnexin-disrupted S. cerevisiae. The mRNA concentrations of BiP and PDI were evidently increased when mutant lysozyme D66H/G49N was expressed in calnexin disrupted S. cerevisiae, while they were not so increased when I55T/G49N mutant was expressed. This observation indicates that the conformation of mutant lysozyme D66H/G49N was less stable in the ER, thus leading to the higher-level expression of ER molecular chaperones via the unfolded protein response pathway. This suggests that glycosylated amyloidogenic lysozyme I55T/G49N may have a relatively stable conformation in the ER, thus releasing it from the quality control of calnexin compared with mutant lysozyme D66H/G49N. PMID- 11852083 TI - Participation of peroxynitrite in oxidative modification of LDL by aqueous extracts of cigarette smoke. AB - Aqueous extracts of cigarette smoke (CSE) can oxidatively modify plasma low density lipoprotein (LDL). The aim of the present study was to elucidate the participation of peroxynitrite in LDL oxidation. When LDL was incubated with CSE, its oxidative modification was dependent on time and concentration. It could be effectively prevented by vitamin E, partially by superoxide dismutase, but hardly by catalase, mannitol and metal chelators. CSE also increased the 3-nitrotyrosine content in LDL. A similar increase of 3-nitrotyrosine occurred after incubation of LDL with a peroxynitrite generating agent, 3-morpholinosydnonimine, thus suggesting that prominent pro-oxidants in CSE are peroxynitrite-generating species. PMID- 11852084 TI - The characterization of the human Siah-1 promoter(1). AB - Siah-1, the human homologue of Drosophila seven in absentia, is related to apoptosis and tumor suppression. Although it was reported that the expression of Siah-1 is induced by p53 and p21/WAF1, little is known about the transcriptional regulation of the Siah-1 gene. To investigate the transcriptional regulation, we isolated and sequenced the genomic fragment of the Siah-1 promoter region. The Siah-1 promoter has no typical TATA box or CCAAT box. Transient transfection assays using reporter plasmids in which the promoter region of the Siah-1 gene was deleted or mutated showed that one Sp1 site was responsible for the basal promoter activity. In Northern blotting analysis, the expression of the Siah-1 gene was upregulated by p53, but activation of the reporter plasmid by the p53 co transfection assay was not shown, suggesting that a p53 responsive element does not exist in the promoter region we examined in this study but might be present in another region. PMID- 11852085 TI - A quantity control mechanism regulating levels of the HapE subunit of the Hap complex in Aspergillus nidulans: no accumulation of HapE in hapC deletion mutants. AB - The Aspergillus nidulans CCAAT-binding complex (Hap complex) consists of at least three subunits, HapB, HapC and HapE. To investigate the quantity control mechanisms of the subunits during assembly of the Hap complex, reconstitution studies with the recombinant subunits and extracts prepared from the respective hap subunit deletion mutants were carried out. Furthermore, Western blot analysis of the Hap subunits and Northern blot analysis of the hap genes with the respective deletion mutants were also performed. From all the results together, it was suggested that the number of the HapC molecule could adjust that of the HapE molecule by forming stable heterodimers prior to assembly of the Hap complex. PMID- 11852086 TI - Molecular cloning and characterization of Kv6.3, a novel modulatory subunit for voltage-gated K(+) channel Kv2.1. AB - We report identification and characterization of Kv6.3, a novel member of the voltage-gated K(+) channel. Reverse transcriptase-polymerase chain reaction analysis indicated that Kv6.3 was highly expressed in the brain. Electrophysiological studies indicated that homomultimeric Kv6.3 did not yield a functional voltage-gated ion channel. When Kv6.3 and Kv2.1 were co-expressed, the heteromultimeric channels displayed the decreased rate of deactivation compared to the homomultimeric Kv2.1 channels. Immunoprecipitation studies indicated that Kv6.3 bound with Kv2.1 in co-transfected cells. These results indicate that Kv6.3 is a novel member of the voltage-gated K(+) channel which functions as a modulatory subunit. PMID- 11852087 TI - Imaging of the fluorescence spectrum of a single fluorescent molecule by prism based spectroscopy. AB - We have devised a novel method to visualize the fluorescence spectrum of a single fluorescent molecule using prism-based spectroscopy. Equipping a total internal reflection microscope with a newly designed wedge prism, we obtained a spectral image of a single rhodamine red molecule attached to an essential light chain of myosin. We also obtained a spectral image of single-pair fluorescence resonance energy transfer between rhodamine red and Cy5 in a double-labeled myosin motor domain. This method could become a useful tool to investigate the dynamic processes of biomolecules at the single-molecule level. PMID- 11852088 TI - Characterization of a bacterial tyrosine ammonia lyase, a biosynthetic enzyme for the photoactive yellow protein. AB - During genome sequence analysis of Rhodobacter capsulatus, nearby open reading frames were found that encode a photoactive yellow protein (PYP) and a hypothetical biosynthetic enzyme for its chromophore, a tyrosine ammonia lyase (TAL). We isolated the TAL gene, overproduced the recombinant protein in Escherichia coli, and after purification analyzed the enzyme for its activity. The catalytic efficiency for tyrosine was shown to be approximately 150 times larger than for phenylalanine, suggesting that the enzyme could in fact be involved in biosynthesis of the PYP chromophore. To our knowledge it is the first time this type of enzyme has been found in bacteria. PMID- 11852089 TI - Aggregation of LDL with chondroitin-4-sulfate makes LDL oxidizable in the presence of water-soluble antioxidants. AB - The content of plasma and arterial interstitial fluid in water-soluble antioxidants makes it unlikely for low-density lipoprotein (LDL) to oxidize by the oxidation mechanisms most frequently discussed. By aggregation of LDL in the presence of chondroitin-4-sulfate (C-4-S), but not with chondroitin-6-sulfate or sphingomyelinase, a complex arises which can oxidize in the presence of 20 microM ascorbate and 300 microM urate. This oxidation sensitivity even persists after the gel-filtration of an LDL/C-4-S/Cu(2+) complex, indicating entrapment of Cu(2+) within. This corresponds well to the known ability of C-4-S to bind copper ions and is a potential mechanism by which LDL oxidation in the arterial intima is facilitated after prolonged retention by the extracellular matrix. PMID- 11852090 TI - The glycan core of GPI-anchored proteins modulates aerolysin binding but is not sufficient: the polypeptide moiety is required for the toxin-receptor interaction. AB - Sensitivity of mammalian cells to the bacterial toxin aerolysin is due to the presence at their surface of glycosylphosphatidyl inositol (GPI)-anchored proteins which act as receptors. Using a panel of mutants that are affected in the GPI biosynthetic pathway and Trypanosoma brucei variant surface glycoproteins, we show that addition of an ethanolamine phosphate residue on the first mannose of the glycan core does not affect binding. In contrast, the addition of a side chain of up to four galactose residues at position 3 of this same mannose leads to an increase in binding. However, protein free GPIs, which accumulate in mutant cells deficient in the transamidase that transfers the protein to the pre-formed GPI-anchor, were unable to bind the toxin indicating a requirement for the polypeptide moiety, the nature and size of which seem of little importance although two exceptions have been identified. PMID- 11852091 TI - A novel rat hypothalamic RFamide-related peptide identified by immunoaffinity chromatography and mass spectrometry. AB - Recently, cDNAs encoding novel RFamide-related peptides (RFRPs) have been reported in the mammalian brains by a gene database search and the deduced RFRPs have been suggested to participate in neuroendocrine and pain mechanisms in the rat. Two peptides have been predicted to be encoded in the cDNA of rodent RFRPs. To assess precise functions of rodent RFRPs in the brain, in the present study we identified a naturally occurring RFRP in the rat hypothalamus by immunoaffinity purification combined with mass spectrometry (MS). The affinity chromatography showed that the rat hypothalamus contained RFRP-like immunoreactivity. The immunoreactive material was analyzed by a nanoflow electrospray ionization time of-flight MS followed by tandem MS analysis. The mass peak corresponding to octadecapeptide was detected at 1010.54 m/z ([M+2H](2+)) and its sequence, ANMEAGTMSHFPSLPQRF-NH(2), was revealed by the fragmentation, showing a mature form encoded in the cDNA sequence of RFRPs. The identified endogenous RFRP will aid not only in defining its physiological roles but also facilitate the development of its agonists and antagonists in the rodent brain. PMID- 11852092 TI - ACTH secretion by mouse corticotroph AtT20 cells is negatively modulated by the intracellular level of 7B2. AB - 7B2 is a pan-neuroendocrine protein known to facilitate the trafficking and activation of the prohormone proprotein convertase-2 (PC2). 7B2-null mice not only lack PC2 activity, but they also develop an adrenocorticotropic hormone (ACTH) hypersecretion syndrome, suggesting that 7B2 may regulate hormone secretion. To verify this possibility, we introduced into mouse corticotroph AtT20 cells a retroviral vector carrying either a sense or an antisense 7B2 transgene to induce higher and lower 7B2 expression, respectively. Relative to control AtT20 cells, 7B2-overexpressing cells released less ACTH following KCl induced membrane depolarization, whereas cells expressing lower levels of 7B2 released relatively more, suggesting that 7B2-related peptides modulate regulated secretion in neuroendocrine cells. PMID- 11852094 TI - Characterization of a lysK gene as an argE homolog in Thermus thermophilus HB27. AB - We conducted a chromosome walk to obtain a DNA fragment downstream of lysJ and found an argE homolog in a putative operon composed of lysJ-orfC-orfD-argE homologs. A knockout mutant of the argE homolog showed significantly slow growth on a minimal medium, and the growth was markedly improved by addition of lysine. We therefore termed this gene lysK. Purified LysK protein has deacetylating activities for both N(2)-acetyllysine and N(2)-acetylornithine at almost equal efficiency. These results suggest that lysK which may share an ancestor with argE functions not only for the lysine biosynthesis, but also for arginine biosynthesis in Thermus thermophilus. PMID- 11852093 TI - Heterodisulfide reductase from Methanothermobacter marburgensis contains an active-site [4Fe-4S] cluster that is directly involved in mediating heterodisulfide reduction. AB - Heterodisulfide reductases (HDRs) from methanogenic archaea are iron-sulfur flavoproteins or hemoproteins that catalyze the reversible reduction of the heterodisulfide (CoM-S-S-CoB) of the methanogenic thiol coenzymes, coenzyme M (CoM-SH) and coenzyme B (CoB-SH). In this work, the ground- and excited-state electronic properties of the paramagnetic Fe-S clusters in Methanothermobacter marburgensis HDR have been characterized using the combination of electron paramagnetic resonance and variable-temperature magnetic circular dichroism spectroscopies. The results confirm multiple S=1/2 [4Fe-4S](+) clusters in dithionite-reduced HDR and reveal spectroscopically distinct S=1/2 [4Fe-4S](3+) clusters in oxidized HDR samples treated separately with the CoM-SH and CoB-SH cosubstrates. The active site of HDR is therefore shown to contain a [4Fe-4S] cluster that is directly involved in mediating heterodisulfide reduction. The catalytic mechanism of HDR is discussed in light of the crystallographic and spectroscopic studies of the related chloroplast ferredoxin:thioredoxin reductase class of disulfide reductases. PMID- 11852095 TI - SNARE expression and distribution during 3T3-L1 adipocyte differentiation. AB - Differentiation of 3T3-L1 cells into adipocytes presupposes the expression of the glucose transporter isoform GLUT4 and the acquisition of insulin-dependent GLUT4 translocation from intracellular storage vesicles to plasma membrane. This ability to translocate GLUT4 depends on the presence of a set of proteins of the SNARE category that are essential in the fusion step. The expression and levels of some of these SNARE proteins are altered during 3T3-L1 differentiation. Levels of the v-SNARE protein cellubrevin and of the t-SNARE protein syntaxin 4 were increased in this process in parallel to GLUT4. However, the levels of SNAP-23, another t-SNARE, were maintained during differentiation. Immunofluorescence images of SNAP-23 showed the initial distribution of this protein in a perinuclear region before differentiation and its redistribution towards plasma membrane in the adipocyte form. These results suggest a capital role in the expression levels and cellular distribution, during 3T3-L1 differentiation, of SNARE proteins involved in the late steps of GLUT4 translocation. PMID- 11852096 TI - Tyrosine kinase participates in vasoconstriction through a Ca(2+)- and myosin light chain phosphorylation-independent pathway. AB - This study was undertaken to determine the role of tyrosine kinase on intracellular Ca(2+) ([Ca(2+)](i)), myosin light chain (MLC) phosphorylation, and contraction caused by norepinephrine (NE) in rat aorta. NE induced a sustained contraction with an increase of [Ca(2+)](i). On the other hand, NE increased the phosphorylation of the 20 kDa MLC transiently. Pretreatment with genistein and tyrophostin 25, tyrosine kinase inhibitors, significantly inhibited NE-induced contraction, but did not affect the increase of [Ca(2+)](i) and MLC phosphorylation. These results suggest that tyrosine kinase may regulate the NE mediated contraction without altering [Ca(2+)](i) and MLC phosphorylation in rat aorta. PMID- 11852097 TI - Molecular aspects of photodynamic therapy: low energy pre-sensitization of hypericin-loaded human endometrial carcinoma cells enhances photo-tolerance, alters gene expression and affects the cell cycle. AB - Photosensitization of HEC1-B cells with a low concentration of hypericin and doses of light below 10 J/cm(2) caused cell death (apoptosis occurred mainly at doses between 2 and 5 J/cm(2), whereas necrosis prevailed above 6 J/cm(2)). However, pre-exposure of cells to innocuous irradiation (2 J/cm(2)) and successive challenge with a light dose that normally induced apoptosis (5 J/cm(2)) altered the expression of the proteins involved in the regulation of apoptosis, stress response and cell cycle. This change resulted in a significant increase in cell photo-tolerance. PMID- 11852098 TI - Frataxin knockin mouse. AB - Friedreich ataxia is the consequence of frataxin deficiency, most often caused by a GAA repeat expansion in intron 1 of the corresponding gene. Frataxin is a mitochondrial protein involved in iron homeostasis. As an attempt to generate a mouse model of the disease, we introduced a (GAA)(230) repeat within the mouse frataxin gene by homologous recombination. GAA repeat knockin mice were crossed with frataxin knockout mice to obtain double heterozygous mice expressing 25-36% of wild-type frataxin levels. These mice were viable and did not develop anomalies of motor coordination, iron metabolism or response to iron loading. Repeats were meiotically and mitotically stable. PMID- 11852099 TI - Three-dimensional architecture of the eukaryotic multisynthetase complex determined from negatively stained and cryoelectron micrographs. AB - This study provides the first description of the three-dimensional architecture of the multienzyme complex of aminoacyl-tRNA synthetases. Reconstructions were calculated from electron microscopic images of negatively stained and frozen hydrated samples using three independent angular assignment methods. In all cases, volumes show an asymmetric triangular arrangement of protein domains around a deep central cavity. The structures have openings or indentations on most sides. Maximum dimensions are ca. 19x16x10 nm. The central cavity is 4 nm in diameter and extends two-thirds of the length of the particle. PMID- 11852100 TI - Temperature effects on the presteady-state and transport-associated currents of GABA cotransporter rGAT1. AB - The effects of temperature on the gamma-aminobutyric acid (GABA) uptake and on the presteady-state and transport-associated currents of the GABA cotransporter, rat gamma-aminobutyric acid transporter 1 (rGAT1), have been studied using heterologous oocyte expression and voltage-clamp. Increasing temperature from 15 to 30 degrees C increased GABA uptake, diminished the maximal value of the relaxation time constant of the presteady-state currents and increased the amplitude of the current associated with the transport of GABA. The curve of the presteady-state charge versus voltage was shifted toward negative potentials by increasing the temperature, while the maximal amount of charge (Q(max)) remained constant; the tau versus V curve was also negatively shifted by increasing temperatures. Analysis of the outward (alpha) and inward (beta) rate constants as functions of temperature showed that they are affected differently, with a Q(10)=3.4 for alpha and Q(10)=1.5 for beta. The different temperature coefficients of the rate constants account for the observed shifts. These observations are consistent with a charge moving mechanism based on a conformational change of the protein; the weaker temperature sensitivity of the inward rate constant suggests a rate-limiting diffusional component on this process. PMID- 11852101 TI - Induction of mammalian cell death by a plant Bax inhibitor. AB - Arabidopsis thaliana AtBI-1 is an orthologue of mammalian Bax inhibitor-1 capable of suppressing Bax-induced cell death in yeast as well as mammalian cells. Here we investigated whether or not AtBI-1 suppresses Bax-induced cell death using human fibrosarcoma HT1080 cells. Surprisingly, AtBI-1 did not block Bax-induced cell death, but it triggered apoptotic cell death in mammalian cells. The proapoptotic effect of AtBI-1 was blocked by the X-linked caspase inhibitor XIAP, suggesting that the cell death caused by AtBI-1 is similar to that caused by Bax. PMID- 11852102 TI - The involvement of reactive oxygen species (ROS) and p38 mitogen-activated protein (MAP) kinase in TRAIL/Apo2L-induced apoptosis. AB - To determine the apoptotic signaling pathway which tumor necrosis factor-related apoptosis-inducing ligand (TRAIL/Apo2L) induced, we investigated the contribution of reactive oxygen species (ROS), p38 mitogen-activated protein (MAP) kinase and caspases in human adenocarcinoma HeLa cells. Here we show that upon TRAIL/Apo2L exposure there was pronounced ROS accumulation and activation of p38 MAP kinase, and that activation of caspases and apoptosis followed. Pretreatment with antioxidants such as glutathione or estrogen attenuated TRAIL/Apo2L-induced apoptosis through a reduction of ROS generation and diminished p38 MAP kinase and caspase activation. The p38 MAP kinase inhibitor SB203580 prevented apoptosis through a blockage of caspase activation, although ROS generation was not attenuated. Furthermore, the pan-caspase inhibitor Z-Val-Ala-DL-Asp-fluoromethyl ketone fully prevented apoptosis, while neither ROS accumulation nor p38 MAP kinase activation were affected. Therefore, our results suggest that TRAIL/Apo2L induced apoptosis is mediated by ROS-activated p38 MAP kinase followed by caspase activation in HeLa cells. PMID- 11852103 TI - Hydroxylation of fatty acids by microsomal and reconstituted cytochrome P450 2B1. AB - Understanding the mechanisms by which cytochrome(s) P450 (CYP) discriminate good from poor substrates, and orient them for highly regio- and stereoselective oxidation, has considerable intrinsic and practical importance. Here we present results of a study of fatty acid hydroxylation by CYP2B1 in a reconstituted system and in microsomes from phenobarbital-pretreated rats. The results indicate that 2B1 prefers decanoic acid as the optimum fatty acid substrate (among C(8) C(16)) and that it hydroxylates all positions five or more methylene groups distant from the carboxylate carbon. That hydroxylation does not occur at carbon atoms closer to the carboxyl group than the C(6) position suggests that these carbons may not reach the ferryl oxygen because the carboxyl group is anchored to a specific site at a fixed distance from the heme iron. PMID- 11852104 TI - Property comparison of recombinant amphibian and mammalian allantoicases. AB - Allantoicase is an enzyme involved in uric acid degradation. Although it is commonly accepted that allantoicase is lost in mammals, birds and reptiles, we have recently identified its transcripts in mice and humans. The mouse mRNA seems capable of encoding a functional allantoicase, therefore we expressed the Xenopus and mouse allantoicases (MAlc and XAlc, respectively) in Escherichia coli and characterized the recombinant enzymes. The two recombinant allantoicases show a similar temperature and pH stability but, although XAlc and MAlc share a 54% amino acid identity, they differ in sensitivity to bivalent cations, in substrate affinity and in the level of expression in tissues (as revealed by means of Western blot analysis). We propose that the loss of allantoicase activity in mouse is due to a low substrate affinity and to a reduced expression level of the enzyme. PMID- 11852105 TI - Distinct regions of C-terminus of the high affinity neurotensin receptor mediate the functional coupling with pertussis toxin sensitive and insensitive G proteins. AB - The functional coupling of C-terminally truncated mutants of the high affinity rat neurotensin (NT) receptor (NTS1) was characterized in transfected Chinese hamster ovary cells. On cells expressing NTRDelta372 (truncated NTS1 lacking the entire 52 amino acid C-terminus), NT failed to promote [(35)S]guanosine 5'-[gamma (35)S]triphosphate binding whereas a robust pertussis toxin (PTx) sensitive response was observed in cells expressing a partially truncated receptor (NTRDelta401 lacking the last 23 residues). Similar results were obtained when measuring the ability of NT to induce the production of arachidonic acid. Since neither deletions impaired the NT-induced phosphoinositide hydrolysis, these results indicate that the membrane proximal region of the C-terminus is specifically involved in the functional coupling of the receptor with PTx sensitive G-proteins. This region was also found to be involved in the control of receptor internalization. However, PTx failed to impair internalization, indicating that these two properties are not directly related. PMID- 11852106 TI - Curcumin induces apoptosis in human breast cancer cells through p53-dependent Bax induction. AB - The aim of this study was to determine the mechanisms of curcumin-induced human breast cancer cell apoptosis. From quantitative image analysis data showing an increase in the percentage of cells with a sub-G0/G1 DNA content, we demonstrated curcumin-induced apoptosis in the breast cancer cell line MCF-7, in which expression of wild-type p53 could be induced. Apoptosis was accompanied by an increase in p53 level as well as its DNA-binding activity followed by Bax expression at the protein level. Further experiments using p53-null MDAH041 cell as well as low and high p53-expressing TR9-7 cell, in which p53 expression is under tight control of tetracycline, established that curcumin induced apoptosis in tumor cells via a p53-dependent pathway in which Bax is the downstream effector of p53. This property of curcumin suggests that this molecule could have a possible therapeutic potential in breast cancer patients. PMID- 11852107 TI - Insertion and glycosylation of Pf3-derived membrane proteins in microsomes. AB - To get insight into the insertion mechanism of small newly synthesized single spanning membrane proteins, Pf3 coat protein mutants were constructed with potential glycosylation sites in the N-terminus. Some of these proteins, when synthesized in vitro in the presence of microsomes, became efficiently glycosylated, proving that they insert into the membrane and translocate their N terminus to the lumenal side. Such Pf3 constructs also insert efficiently into Escherichia coli vesicles and even in pure lipid vesicles, suggesting a common mechanism, which might be spontaneous. Glycosylation was sensitive to changes in the amino acid sequence of the N-terminus, suggesting that it depends on the structure of the protein and/or its positioning with respect to the lipid-water interface. PMID- 11852108 TI - PDZ domains: troubles in classification. PMID- 11852110 TI - A second melanotransferrin gene (MTf2) and a novel protein isoform: explanation for the membrane-bound and soluble forms of melanotransferrin? PMID- 11852113 TI - Requirement of c-Jun N terminal kinase and P38 mitogen-activated protein kinase for anti-IgM-induced reduction in mitochondrial membrane potential in CH31 B lymphoma cells. AB - CH31 cells have been used for analysis of B cell tolerance, since engagement of membrane immunoglobulin (mIg) results in loss in mitochondrial membrane potential (DeltaPsim), followed by cell death. We have reported that the dominant-negative (dn) form of c-Jun N-terminal kinase (JNK) substantially prevented a loss of DeltaPsim at 24 h, with partial protection around 48 h after anti-IgM stimulation. In this study, we demonstrate that anti-IgM induced a sustained activation of p38 mitogen-activated protein (MAP) kinase. The p38MAP kinase inhibitor SB203580 substantially prevented loss of DeltaPsim at 14 h, with partial prevention (18-24 h) after anti-IgM stimulation. The dnJNK1-mediated prevention of anti-IgM-induced mitochondrial dissipation was enhanced by SB203580 at 42 h, but not 24 h, after stimulation, suggesting that activation of either p38 MAP kinase or JNK may be sufficient for the initiation of early phase of anti IgM-induced loss of DeltaPsim while both may be necessary in the late phase. PMID- 11852114 TI - Specific and effective targeting cancer immunotherapy with a combination of three bispecific antibodies. AB - For the purpose of establishing a new adoptive immunotherapy for bile duct carcinoma (BDC), we previously constructed two kinds of bispecific antibodies (bsAbs), anti-MUC1 x anti-CD3 (M x 3) and anti-MUC1 x anti-CD28 (M x 28), which activate T cells and form bridges between them and MUC1-expressing tumor cells. In our previous studies [Cancer Res. 56 (1996) 4205] specific targeting therapy (STT) consisting of i.v. administration of lymphokine activated killer cells with a T cell phenotype (T-LAK) sensitized with two kinds of bsAbs to human BDC grafted severe combined immunodeficient (SCID) mice demonstrated remarkable inhibition of tumor growth. However, complete cures could not be obtained. In order to improve antitumor efficacy, we have paid attention to anti-CD2 monoclonal antibodies (mAbs), thought to play an important roles in signal transduction in T cell activation or control of T cell receptor (TCR)-driven activation. Therefore, we developed another bsAb, anti-MUC1 x anti-CD2 (M x 2), in order to examine if this would show synergism with the two previously described bsAbs. The combination of the three bsAbs (M x 3, M x 28 and M x 2 bsAbs) showed highest cytotoxicity against MUC1-expressing BDC cells when given simultaneously with peripheral blood mononuclear cells (PBMCs) or T-LAK cells in vitro. When 2 x 10(7) T-LAK cells sensitized with different combinations of bsAbs were administered four times i.v. to BDC-grafted SCID mice, the best therapeutic result was obtained with a combination of all three bsAbs. These results indicate usefulness of combination of three bsAbs for targeting cancer immunotherapy. PMID- 11852115 TI - The expression of Bcl-2 family proteins (Bcl-2, Bcl-x, Bax, Bak and Bim) in human lymphocytes. AB - Bcl-2 family proteins regulate programmed cell death, and may play an important role in the selection of lymphocytes. We investigated the expression of Bcl-2, Bcl-x, Bax, Bak and Bim in human lymphocytes using flow-cytometry. Bcl-2 was down regulated in CD4(+)8(+) (DP) thymocytes and CD19(+)38(+) tonsillar lymphocytes (GC B cells). Among DP thymocytes, cells co-expressing CD69 up-regulated Bcl-2, suggesting that the role of Bcl-2 is promoting survival of positively selected DP cells. Unexpectedly, the expression level of Bcl-x was higher in DP cells than in Single Positive (SP) cells and in CD69(+) DP thymocytes it was lower than in CD69(+) DP thymocytes. Expression of Bim was low in DP thymocytes but high in a subset of GC B cells. Bim and Bax were expressed more highly in SP than in DP thymocytes. Among peripheral blood lymphocytes (PBL), CD8(+) T cells expressed an approximately ten-fold higher level of Bcl-x than CD4(+) T cells while both subsets expressed similar levels of Bcl-2. Bak expression was low and Bim expression was absent in PBL. These results suggest that not only Bcl-2 but other members of the Bcl-2 family are involved in T cell development in the thymus and affinity maturation of B cells in the germinal center. PMID- 11852116 TI - Culture filtrate specific H-2(b) restricted CD8+ T cells activated in vivo by Mycobacterium tuberculosis or bovis BCG recognize a restricted number of immunodominant peptides. AB - We previously demonstrated that Bacillus Calmette-Guerin (BCG) immunization activated D(b) restricted CD8+ cytolytic T lymphocyte (CTL) recognizing target cells incubated with mycobacterial culture filtrate. Here, we show that in vitro restimulation of spleen cells from BCG vaccinated or Mycobacterium tuberculosis infected mice with culture filtrate antigens leads to the appearance of a high percentage of D(b) restricted IFNgamma synthesizing CD8+ T cell blasts. Transporter associated protein-2 mutated RMA-S cells incubated with soluble culture filtrate proteins had their MHC class I D(b) but not K(b) molecules stabilized at the surface indicating that only D(b) ligands might be generated by antigen presenting cells. MHC class I bound peptides were acid eluted from the surface of RMA-S cells incubated with M. tuberculosis culture filtrate proteins. The crude peptide preparation was able to sensitize RMA-S cells for recognition by culture filtrate-specific cytolytic T cells. Peptides were subsequently fractionnated by reverse-phase high performance liquid chromatography and the main biological activity was identified in two fractions. These results provide a further evidence that the processing of exogenous culture filtrate proteins in vitro leads to the presentation of a restricted number or even a single immunodominant peptide to culture filtrate-specific CD8+ T cells. PMID- 11852117 TI - Sister cytotoxic CD8+ T cell clones differing in natural killer inhibitory receptor expression in human astrocytoma. AB - Natural killer (NK) inhibitory receptors are thought to play a critical role in the regulation of cytotoxicity of NK cells and certain self-reactive T cells. In the present study, we investigated whether astrocytoma infiltrating T lymphocytes may be functionally compromised by NK receptors (NKRs). The NK inhibitory receptor CD94/NKG2A was found on a significant proportion of CD8+ astrocytoma infiltrating lymphocytes. The functional consequences of CD94/NKG2A expression were explored at the clonal level, using a T cell clone that exhibited substantial variation in the expression of this heterodimer. Triggering of CD94/NKG2A inhibited the killing properties of T cells with a high level of this receptor, but not those from T cells with a low level. Our data indicate that some astrocytoma infiltrating lymphocytes express functional inhibitory CD94/NKG2A, raising the possibility that they may represent silent T cells specific for self-antigens (Ags) expressed on tumor cells. Understanding the mechanisms of regulation of these receptors may bring new insights for optimizing an anti-tumor immune response. PMID- 11852118 TI - Lymphocyte phenotype analysis and chromosome aberration frequency of workers occupationally exposed to styrene, benzene, polycyclic aromatic hydrocarbons or mixed solvents. AB - The aim of our study was to investigate the immunotoxicity of benzene, styrene and polycyclic aromatic hydrocarbon exposure, to establish the correlation between immunological and genotoxicological parameters, and to assess the possible effect of confounding factors such as age and smoking. The immune status of the donors was characterized by measuring the surface antigens of peripheral lymphocytes. The studied antigens were the following: CD3, CD4, CD8, CD14, CD19, CD25, CD38, CD45, CD45RO, CD54, CD56, CD62L, CD71 and HLA-DR. In our studies, we compared the immunological and genotoxicological parameters (chromosome aberration, sister chromatid exchange frequency, unscheduled DNA synthesis) of the different groups with healthy controls. Analysis revealed changes in the expression of surface antigens on peripheral lymphocytes in correlation with exposure. Confounding factors, such as smoking, increased the proportion of CD4 positive T lymphocytes and influenced the surface expression of several antigens. In our investigation the occurrence of chromosome aberrations negatively correlated with CD25 (IL-2R) expression in both CD4 and CD8 positive T cells. The presented data suggest that solvents such as benzene, styrene and PAHs activate peripheral lymphocytes, and cause changes in the incidence of CD25+/CD4+ T lymphocytes that may represent a distinct subset of immune-regulatory T cells. PMID- 11852119 TI - Polyclonal anti-colorectal cancer Fab phage display library selected in one round using density gradient centrifugation to separate antigen-bound and free phage. AB - A combinatorial Fab phage display library generated from antibody variable (V) region genes of BALB/c mice immunized with the human colorectal cancer cell lines SW480, SW948, and SW837, was used to isolate an anti-colorectal cancer library. In an attempt to preserve as many anti-colorectal cancer specificities as possible, the original Fab phage display library was selected for binding to a suspension of the human colorectal cancer cells using density gradient centrifugation, instead of washes, to separate cell-bound and free phage. The method consists of placing the cell-phage mixture on a layer of fetal bovine serum (FBS) which had been overlaid on a "cushion" of percoll density medium in a soft, polyallomer tube. After centrifugation, free phage remain on top of the serum layer, whereas the colorectal cancer cells with bound phage are recovered from the serum-percoll interface with a syringe. Analysis of the selected phage display library by enzyme linked immunosorbent assay (ELISA) and diagnostic restriction enzyme digests of individual members (fingerprinting), revealed about 90% anti-colorectal cancer diverse clones after only one round of selection. After conversion to a library of full-length antibodies, such an anti-cancer polyclonal library could be useful for therapeutic and/or diagnostic applications. The density gradient centrifugation method presented here holds great promise for the generation of polyclonal antibody libraries (PCALs) to complex antigens. It is also applicable for selection of peptide or other phage display libraries on any insoluble ligand. PMID- 11852120 TI - Cross-linking HLA-DR molecules on Th1 cells induces anergy in association with increased level of cyclin-dependent kinase inhibitor p27(Kip1). AB - HLA class II molecules play pivotal roles in antigen presentation to CD4+ T cells. We investigated signaling via HLA-DR molecules expressed on CD4+ T cells. When HLA-DR or CD3 molecules on cloned CD4+ T cells were cross-linked by solid phase mAbs, T cells proliferated, and this resulted in anergy. Whereas cross linking of HLA-DR and CD3 resulted in secretion of the same levels of IFN-gamma and IL-8, secretion of IL-10 induced by cross-linking of HLA-DR was less than that induced by cross-linking of CD3 on CD4+ T cells. Interestingly, expression of p27(Kip1) but not p21(Cip1) increased after stimulation by either anti-HLA-DR or anti-CD3 mAb. This was indeed the case, when T cells were rendered anergic using a soluble form of antigenic peptide. In contrast, T cells stimulated by peptide-pulsed PBMC expressed little p27(Kip1). We propose that signaling via HLA DR molecules on CD4+ T cells at least in part contributes to the induction of T cell anergy, through the upregulated expression of the p27(Kip1). The implication of our finding is that HLA-DR molecules play a role in human T cell anergy induced by a soluble form of antigenic peptide. PMID- 11852121 TI - New variant CJD-BSE (mad cow disease). The need for disposable ENT instruments. AB - This paper outlines the development of Bovine Spongiform Encephalopathy (BSE) in the United Kingdom. The relationship between BSE and new variant Creutzfeldt Jakob disease (vCJD) is considered and the risks of iatrogenic spread reviewed. The rationale for disposable surgical instruments in adenotonsillectomy to prevent iatrogenic spread is discussed. PMID- 11852122 TI - The role of laser assisted tympanostomy (LAT) in treating allergic children with chronic serous otitis media. AB - OBJECTIVES: To assess various prognostic factors influencing the outcome in paediatric patients with serous otitis media, who have undergone laser assisted tympanostomy without ventilation tube placement. Emphasis is given to children with allergies who underwent the procedure. METHOD: Laser assisted tympanostomy was performed on a total of 130 ears (92 individuals) with chronic otitis media with effusion. To determine the quality of patient outcome, the following parameters were evaluated: external auditory canal anatomy, type of anaesthesia used, tympanic membrane and middle ear fluid characteristics, myringotomy size, a history of allergies and the laser device parameters. RESULTS: Multivariable statistical analysis demonstrated that the presence of allergies in children with chronic serous otitis media is significantly correlated with a poor outcome (P < 0.0047). Moreover, the presence of a thick tympanic membrane and/or high viscosity (glue) fluid in the middle ear cavity can also independently influence patient outcome (P < 0.025). Simultaneous adenoidectomy and/or tonsillectomy, type of anaesthesia (general versus local), external canal anatomy (wide or narrow) and sex, were not statistically important prognostic factors. The type of anaesthesia used, myringotomy size and the laser device parameters were not found to be associated with patient outcome. CONCLUSION: A history of allergies, the presence of a thick tympanic membrane and/or high viscosity fluid in the middle ear cavity are all contraindications for laser assisted tympanostomy without tympanostomy tubes, in children who suffer from chronic serous otitis media. The selection criteria for this procedure in the paediatric population are addressed in detail. PMID- 11852123 TI - Epidemiology of chronic suppurative otitis media among Saudi children--a comparative study of two decades. AB - An epidemiological survey to study the prevalence of chronic suppurative otitis media (CSOM) was completed in February 2000. About 9540 children aged up to 12 years were examined clinically by an otolaryngologist. All 125 children (1.3%) suffered from CSOM, nine with cholesteatoma. The prevalence of CSOM in the Central province of the Kingdom was found to be 1.15%. During the first study done in 1982, the prevalence was 5.5% among 293 children aged 6-18 years. In the second study done and completed in 1991 where 6421 children were surveyed, 94 children found to suffer from CSOM i.e. 1.5% with six children having cholesteatoma. These results showed a decrease in incidence of CSOM in the Central province, although in other parts of the Kingdom, the prevalence was found to be as high as 2.89%. The factors predisposing to or associated with chronic ear disease will be discussed, and the results will be compared with results published from other centers. PMID- 11852124 TI - Epidemiology of acute otitis media among Saudi children. AB - An epidemiological survey was carried out with 9540 children aged up to 12 years to study the prevalence of acute suppurative otitis media and the rate of hearing impairment. A total of 100 (1.05%) were diagnosed with acute otitis media (AOM). The incidence was found to be higher among young children up to 4-years-old and lower in the age group 8-12 years. The male children had slightly higher rate of AOM as compared with female (1.36 and 0.80%), respectively, (P < 0.01). The prevalence of AOM in the children from the different provinces varied, being higher in those from the Southern and Central regions. Also it was found that the prevalence of ASOM was higher among children whose parents were cousins compared with non-relative parents (1.38 and 0.74%) (P > 0.001). Those with poor socio economic condition showed higher rate especially those living in the Southern part with poor or inadequate health services provided. ASOM was significantly associated with hearing impairment (P < 0.00001). PMID- 11852125 TI - The role of age in pediatric cochlear implantation. AB - OBJECTIVE: To document progress, benefit and importance of age in paediatric cochlear implantation. DESIGN: The EARS (Evaluation of Auditory Responses to Speech) test battery was performed on 33 prelingually deaf children at regular intervals up to 36 months following implantation. All children participated in individually tailored intensive audiological rehabilitation programs after receiving their implants. In this respect, it was attempted to evaluate speech perception scores in children implanted before and after the age of 3 in a homogenous group. RESULTS: All children demonstrated encouraging improvements over time in their speech recognition abilities. Furthermore, it was observed that the children who were implanted under the age of 3 achieved higher levels of speech perception performance. CONCLUSION: In order to shorten the process of central maturation of the auditory system, it is desirable to implant the children as young as possible. Early intervention seems to be the ideal strategy in enabling prelingually deaf children to derive maximum benefit from cochlear implantation. PMID- 11852126 TI - Uvulopalatopharyngoplasty with tonsillectomy and adenoidectomy as a treatment for obstructive sleep apnea in neurologically impaired children. AB - RATIONALE: Children with neurologic impairment often present with airway obstruction that may require intervention. No single method of airway intervention is universally appropriate and effective in this patient population. This study was performed to examine the effectiveness of using adenotonsillectomy and uvulopalatopharyngoplasty (UPPP) in resolving obstructive apnea (OA) in patients with neurologic impairment. METHODS: A retrospective chart review of 15 patients with neurologic impairment and OA treated with adenotonsillectomy and UPPP between 1986 and 1998 at Children's Hospital of Wisconsin (CHW) was performed. All patients in the series had their primary area of obstruction in the posterior oropharynx involving the soft palate, pharyngeal walls and base of tongue. Post-operative improvement following adenotonsillectomy and UPPP was examined. Measures of improvement were based primarily on recorded lowest oxygen saturations, but clinical parameters, flexible upper airway endoscopy and polysomnography were used as well. RESULTS: Patient improvement was documented in 87% of patients treated with this modality. For the group, the mean lowest recorded oxygen saturation demonstrated a statistically significant improvement from 65% pre-operatively to 85% post-operatively (P = 0.005). In long-term follow up of these patients, 77% (10 of 13) of those showing initial improvement have done well and have required no further airway intervention. However, 23% of these patients demonstrated the need for further airway intervention during follow-up. CONCLUSIONS: Adenotonsillectomy with UPPP is worthy of consideration in certain neurologically impaired patients with moderate to severe OA, limited primarily to the posterior pharyngeal area. Initial improvement may not be permanent and close long-term follow-up of patients is imperative. PMID- 11852127 TI - Acute mastoiditis in a Norwegian population: a 20 year retrospective study. AB - We have retrospectively examined the nature of acute mastioditis (in western Norway) during a 20 year period (1980-2000). Sixty-one cases of AM were identified in 57 patients with a mean age of 3.6 years. We found no significant change in the incidence of AM during the last 20 years. Seven patients were treated solely with intravenous antibiotics and myringotomies. Fifty patients also underwent cortical mastoidectomy, four cases with bilateral surgery. Antibiotic treatment was given to 31 of the patients before admission to hospital and this group had a significant longer duration of symptoms (12.4 days) compared to untreated patients (7.3 days). Streptococcus pneumoniae was the most common organism recovered from patient cultures. Surgery was found to correlate to patients with retroauricular fluctuation or to children with at least two of the three clinical signs: protrusion of the ear, retroauricular oedema and swelling of the ear canal. Our data show that clinical examination only reveal 50% of the cases with surgically proven retroauricular subperiostal abscess. We therefore recommend a CT scan of patients treated conservatively. PMID- 11852128 TI - Clinical characteristics of aero-digestive foreign bodies in neurologically impaired children. AB - OBJECTIVE: To determine if children with neurological impairment are at additional risk for foreign body aspiration or impaction and if they present with a different clinical course than normal children. DESIGN: Retrospective cohort study. SETTING: Tertiary care children's hospital. PATIENTS AND OTHER PARTICIPANTS: A retrospective chart review of 328 consecutive procedures for aero digestive foreign body removal over a 6 year period was performed. Of these patients 52 (15.9%) were identified with neurological impairment. RESULTS: Compared with the non-impaired children, those with neurological impairment were older, diagnosed later, hospitalized longer and had a higher incidence of complications. CONCLUSION: A greater index of suspicion is needed to timely treat aero-digestive foreign bodies in children with neurological impairment. These children may not be receiving the necessary attention when presenting with non specific symptoms. PMID- 11852129 TI - Unilateral choanal atresia in identical twins: case report and literature review. AB - Choanal atresia is postulated to be secondary to an abnormality of the rupture of the buccopharyngeal membrane during the embryological period. This condition usually occurs sporadically, but has been described in siblings and successive generations. The genetics remain unclear. We present monozygotic twins with identical findings of unilateral choanal atresia and no other associated anomalies. To our knowledge, this is the first report of such an occurrence. PMID- 11852130 TI - Retrieval of bronchial foreign body with central lumen using a flexible bronchoscope. AB - A 5 year-old boy aspirated a metal spring accidentally. The right bronchial foreign body was removed by threading the alligator of a biopsy forceps through the lumen of the spring while opening its jaws distally. This case illustrates that a foreign body with a central lumen in a child can be successfully removed using a flexible bronchoscope, thus obviating the need for general anesthesia and rigid bronchoscopy. PMID- 11852131 TI - A case of fatal pharyngitis. AB - Pharyngitis is a common condition and usually does not lead to any major complication. We report a case in which the patient presented with pharyngitis resulting in an unusual fatal complication of pharyngeal and cerebral abscess. The causative organism was Streptococcus milleri that should be readily treatable by penicillin if the condition was recognised earlier. PMID- 11852132 TI - Postlingual collapse of language and its recovery after cochlear implantation. AB - A 6-year-old boy lost his normally-developed language ability within 2 months after bilateral sudden peripheral deafness. The boy became non-communicative with others, restless with frequent meaningless bursts of shouts (communication skills equivalent to 9-11 months of language development: a rapid breakdown of language). Since conservative methods were ineffective, cochlear implantation was performed. A surprising success was observed: he regained the language retracing the normal developmental stages and caught up with his contemporaries in 2 years. Behavioral improvement paralleled his language development. This case (1) provides insights into the brain function with respect to language acquisition, in relation to the plasticity during the 'critical period' of language learning, (2) reveals the close relationship between language development and behavior, and (3) suggests the predominance of auditory stimulation in learning language. PMID- 11852133 TI - Towards the comprehension of genetic mechanisms controlling brain morphogenesis. AB - Over the past few years, a huge amount of work has provided mouse mutants for many genes required for regionalization of the developing brain. This remarkable work now offers the opportunity of unmasking new and unexpected gene functions that underlie a complex network of molecular interactions. PMID- 11852134 TI - '...and C is for Clioquinol' - the AbetaCs of Alzheimer's disease. AB - Alzheimer's disease (AD) is a devastating age-related neurodegenerative disorder that has been intensively studied over the last several years. In vitro and in vivo studies have led to an understanding of some of the physico-chemical properties of amyloid, a well-characterized hallmark of AD. Clioquinol is a drug that acts on amyloid by perturbing amyloid's metallo-chemistry, and Clioquinol treatment has been shown to be beneficial in a mouse model of AD. This short review examines the recent studies relating to Clioquinol and AD, and anticipates the imminent results of a Phase II trial of Clioquinol in AD, due in March 2002. PMID- 11852136 TI - Rhodopsin phosphorylation: from terminating single photon responses to photoreceptor dark adaptation. AB - Rhodopsin phosphorylation has provided one of the first examples of the ubiquitous regulatory pattern of specific kinases downregulating the activity of G-protein-coupled receptors. However, only recently have studies in living animals allowed us to consider the role of rhodopsin phosphorylation in a broader spectrum of visual functions, ranging from the ability of rods to generate reproducible electrical responses to their ability to adapt to darkness after substantial light exposures. PMID- 11852137 TI - Autoimmunity and inflammation in the peripheral nervous system. AB - Peripheral Nerve Society Biennial Meeting, organized in conjunction with the Austrian Federal Ministry of Education, Research and Culture, held at Telfs, Tyrol, Austria from 8-12 September 2001. PMID- 11852138 TI - Developing concepts in neural stem cells. AB - Stem Cells in the Mammalian Brain: the 4th Brain Research Interactive Symposium, at the 2001 Annual Conference of the Society for Neuroscience, San Diego, CA, USA from November 8-10 2001. PMID- 11852139 TI - Of layers and spheres: the reaggregate approach in tissue engineering. AB - The reaggregate approach involves the regeneration of histotypical three dimensional spheres from dispersed cells of a given tissue in suspension culture. Reaggregated spheres are used as tumour, genetic, toxicological, biohybrid and neurosphere models, and often replace animal experimentation. A particularly instructive example is the use of reaggregation to regenerate complete laminar tissue from avian embryonic retina. By revealing constraints of layered tissue formation, such retinal spheres could be instrumental for regenerative medicine, including stem cell-based tissue engineering. PMID- 11852140 TI - Tuning of cortical neurons to behaviorally salient acoustic signals. PMID- 11852141 TI - Neurogenetics: white matter matters. PMID- 11852142 TI - Afraid of complications in gene targeting? Anxiety plays a role! PMID- 11852144 TI - Amyloid beta protein forms ion channels. PMID- 11852145 TI - Mossy cells in epilepsy: rigor mortis or vigor mortis? AB - Mossy cells are bi-directionally connected through a positive feedback loop to granule cells, the principal cells of the dentate gyrus. This recurrent circuit is strategically placed between the entorhinal cortex and the hippocampal CA3 region. In spite of their potentially pro-convulsive arrangement with granule cells, mossy cells have not been seriously considered to promote seizures, because mossy cells, allegedly one of the most vulnerable cell types in the entire mammalian brain, have long been 'known' to die en masse in epilepsy. However, new data suggest that rumors of the rapid demise of the mossy cells might have been greatly exaggerated. PMID- 11852146 TI - Ephrin-As as receptors in topographic projections. AB - The Eph family of receptor tyrosine kinases and their 'ligands', the ephrins, have been implicated in a large number of developmental processes, such as boundary formation, cell migration, axon guidance and vasculogenesis. A characteristic of the EphB subclass is that both EphBs and transmembrane-anchored ephrin-Bs function as receptors and as ligands, a phenomenon commonly described as 'bi-directional signalling'. Here we review recent data indicating that EphA receptors and glycosylphosphatidylinositol (GPI)-anchored ephrin-As can also mediate bi-directional signalling. Moreover, characterization of the expression of ephrin-As on axons of the retinotectal and vomeronasal projections suggests that the EphA subfamily is involved in both repulsive and attractive guidance mechanisms during establishment of neuronal connections. PMID- 11852147 TI - Small is not beautiful: antagonizing functions for the prion protein PrP(C) and its homologue Dpl. AB - A conformational variant of the normal prion protein PrP(C) is believed to be identical to PrP(Sc), the agent that causes prion diseases. Recently, a novel protein, named Doppel (Dpl), was identified that shares significant biochemical and structural homology with PrP(C). In specific strains of PrP(C)-deficient mouse lines, Dpl is overexpressed and causes a neurological disease. Dpl neurotoxicity is counteracted and prevented by PrP(C), but the mechanism of antagonistic PrP(C)-Dpl interaction remains elusive. In contrast to its homologue PrP(C), initial studies suggest that Dpl is dispensable for prion disease progression and for the generation of PrP(Sc). Although we are only beginning to understand its function, the discovery of Dpl has already provided some answers to long-standing questions and is transforming our understanding of prion biology. PMID- 11852148 TI - Cytokine-induced sickness behaviour: mechanisms and implications. AB - Sickness behaviour represents the expression of the adaptive reorganization of the priorities of the host during an infectious episode. This process is triggered by pro-inflammatory cytokines produced by peripheral phagocytic cells in contact with invading micro-organisms. The peripheral immune message is relayed to the brain via a fast neural pathway and a slower humoral pathway, resulting in the expression of pro-inflammatory cytokines in macrophage-like cells and microglia in the brain. The cellular and molecular components of this previously unsuspected system are being progressively identified. These advances are opening new avenues for understanding brain disorders, including depression. PMID- 11852150 TI - Talk to the people. AB - Recent opinion surveys suggest that the majority of the public is willing to accept the use of animals in research if high standards of welfare and effective regulation are in place. The public appears unaware that such standards have existed for some time. The scientific community should now re-focus its communication efforts to include the ethics and animal welfare aspects of this issue. PMID- 11852149 TI - Location, location, location: a spatial view of neurotrophin signal transduction. AB - Neurotrophins were originally identified as target-derived factors that regulate the survival and differentiation of innervating neurons. However, neurotrophins can also be released by presynaptic cells to stimulate postsynaptic neurons. Recent studies indicate that differences exist between the signaling pathways activated by neurotrophin stimulation of nerve terminals (retrograde signaling) and neurotrophin stimulation of cell bodies. Retrograde signaling relies on the formation of signaling endosomes, vesicles containing activated Trk receptors and their ligands. Signaling endosomes travel from the nerve terminals to remote cell bodies, where they selectively activate a novel MAP kinase, Erk5, as well as PI3 kinase, and thereby stimulate neuronal survival. The differences in the signaling pathways activated by neurotrophins, which depends on the location of stimulation, provide a mechanism by which neurons can interpret the 'where' as well as the 'what' of growth factor stimulation. PMID- 11852154 TI - Shifts in blood volume alter the perception of posture: further evidence for somatic graviception. AB - This article examines the relation between the perception of one's own body position and the distribution of fluid along the subject's spinal (z-) axis. Two experiments are reported. The first one is a replication of the Vaitl et al. study [J. Psychophysiol. 27 (1997) 99] which has shown that changes in shifts of blood volume into or out of the thoracic cavity induced by lower body positive pressure (LBPP: +30 mmHg) or lower body negative pressure (LBNP: -30 mmHg) exerted on the lower body led subjects to feel tilted head-up or head-down, respectively. The second experiment was designed to differentiate between the influence of the otoliths and of the changes in fluid distribution on the perception of body position by means of a sled centrifuge in combination with LBPP and LBNP. In both experiments, changes in blood distribution within the thoracic cavity were measured by impedance plethysmography. Forty healthy volunteers (17 females) participated in experiment 1. They were positioned on the side (right-ear-down head position) on a tiltable board which the subject and the experimenter could tilt via remote control around the subjects' z-axis. Subjects were asked to rotate the board until they felt they were in a horizontal posture. The results clearly show that the perception of posture is influenced by the shift in blood distribution. During LBNP subjects perceived being tilted head-up, whereas LBPP led them feel tilted head-down. Thus, the results obtained in the 1997 study were replicated. Fourteen males volunteered in experiment 2. They were positioned on the sled on a centrifuge in the same manner as in experiment 1. The sled could be moved via remote control by both the subject and the experimenter. While the centrifuge rotated (omega=2 pi times 0.6 rotations per second) the subjects were asked to move the sled until they felt they were in a horizontal position. As in experiment 1, shifts in blood volume were induced by LBPP and LBNP. The distance between the binaural axis (position of the otoliths) and the centrifuge axis served as dependent measure indicating the subjective horizontal position. Due to the additional centrifugal forces exerted on the body the shifts in blood volume were more pronounced than in experiment 1 where only gravitational forces were produced. The changes in the perception of posture were influenced by both the otoliths and the fluid distribution in such a way that both interact in a compensatory manner. These results again corroborate the evidence that afferent inputs from the cardiovascular system play a major role in the perception of the body posture. This phenomenon of graviception needs to be further elucidated with respect to the origins of the afferent inputs and the site and type of graviceptors (mechanoreceptors) involved. PMID- 11852155 TI - Selective attention to a facial feature with and without facial context: an ERP study. AB - The present experiment addressed the question whether selectively attending to a facial feature (mouth shape) would benefit from the presence of a correct facial context. Subjects attended selectively to one of two possible mouth shapes belonging to photographs of a face with a happy or sad expression, respectively. These mouths were presented randomly either in isolation, embedded in the original photos, or in an exchanged facial context. The ERP effect of attending mouth shape was a lateral posterior negativity, anterior positivity with an onset latency of 160-200 ms; this effect was completely unaffected by the type of facial context. When the mouth shape and the facial context conflicted, this resulted in a medial parieto-occipital positivity with an onset latency of 180 ms, independent of the relevance of the mouth shape. Finally, there was a late (onset at approx. 400 ms) expression (happy vs. sad) effect, which was strongly lateralized to the right posterior hemisphere and was most prominent for attended stimuli in the correct facial context. For the isolated mouth stimuli, a similarly distributed expression effect was observed at an earlier latency range (180-240 ms). These data suggest the existence of separate, independent and neuroanatomically segregated processors engaged in the selective processing of facial features and the detection of contextual congruence and emotional expression of face stimuli. The data do not support that early selective attention processes benefit from top-down constraints provided by the correct facial context. PMID- 11852156 TI - Hemispheric asymmetries for visual and auditory temporal processing: an evoked potential study. AB - Lateralization for temporal processing was investigated using evoked potentials to an auditory and visual gap detection task in 12 dextral adults. The auditory stimuli consisted of 300-ms bursts of white noise, half of which contained an interruption lasting 4 or 6 ms. The visual stimuli consisted of 130-ms flashes of light, half of which contained a gap lasting 6 or 8 ms. The stimuli were presented bilaterally to both ears or both visual fields. Participants made a forced two-choice discrimination using a bimanual response. Manipulations of the task had no effect on the early evoked components. However, an effect was observed for a late positive component, which occurred approximately 300-400 ms following gap presentation. This component tended to be later and lower in amplitude for the more difficult stimulus conditions. An index of the capacity to discriminate gap from no-gap stimuli was gained by calculating the difference waveform between these conditions. The peak of the difference waveform was delayed for the short-gap stimuli relative to the long-gap stimuli, reflecting decreased levels of difficulty associated with the latter stimuli. Topographic maps of the difference waveforms revealed a prominence over the left hemisphere. The visual stimuli had an occipital parietal focus whereas the auditory stimuli were parietally centered. These results confirm the importance of the left hemisphere for temporal processing and demonstrate that it is not the result of a hemispatial attentional bias or a peripheral sensory asymmetry. PMID- 11852157 TI - Temporal stability of psychophysiological response patterns: concepts and statistical tools. AB - The specificity or patterning of psychophysiological responses to behavioral stimuli is of importance for basic and applied psychophysiological research. However, the statistical tools used to describe specificity are heterogenious and the attempts to describe temporal stability or reliability of such response profiles are even more complicated. The aim of this paper is to present an approach designed to clarify the relationships between different measures of temporal stability of specificity. This is done with a compilation of statistical measures based on variance components. This approach incorporates features of response specificity and reliability. Individual, situational and motivational response specificity are defined in a uniform way with respect to temporal stability. The use of the formulae is demonstrated with an empirical example. As is found in the research on single parameters, the profile reliability of level scores is higher than that of change scores. Furthermore, the stability of individual response patterns is dependent upon the question of whether the data is aggregated across situations or not. Situational response patterns are stable when the data is aggregated across subjects, whereas the stability is low without such aggregation. Issues of inconsistencies of variance estimates are addressed, and the relationships between temporal stability, consistency and covariation of physiological variables are discussed. PMID- 11852158 TI - Time-dependent cortical asymmetries induced by emotional arousal: EEG analysis of event-related synchronization and desynchronization in individually defined frequency bands. AB - Event-related desynchronization (ERD) and synchronization (ERS) in the individually defined theta, alpha-1, alpha-2 and alpha-3 frequency bands were measured in 20 healthy subjects in response to International Affective Picture System (IAPS) stimuli with low, moderate and high arousal (LA, MA and HA) content. The 62-channel EEG, skin conductance response (SCR) and heart rate (HR) were simultaneously recorded while subjects viewed sequentially presented pictures and subjectively rated them after each presentation. In the theta band, both MA and HA vs. LA stimuli induced larger synchronization over the left anterior and bilaterally over posterior cortical leads. However, rather unexpectedly, both MA and HA vs. LA stimuli yielded larger alpha-1 synchronization, predominantly over occipital leads. In both theta and alpha-1 bands, affectively salient stimuli prompted larger ERS against the background of the overall dominance in power synchronization of posterior regions of the right hemisphere, irrespective of stimulus category. Finally, in the alpha-3 band, HA stimuli induce a lateralized time-dependent power increase over anterior leads of the left hemisphere. The hemispheric asymmetries revealed point to recruitment of not only posterior regions of the right hemisphere (theta and alpha-1 bands), but also of anterior regions of the left hemisphere (theta and alpha-3 bands) in affect analysis beyond valence dimension. In terms of affective chronometry, the significant arousalxtime interactions clearly indicate that in the theta frequency band discrimination of affective stimuli has already started at 200 ms post-stimulus, whereas in the alpha-1 and alpha-3 bands this process is delayed by up to 800-1200 ms. PMID- 11852159 TI - Impaired attention modulation of the blink reflex R3 component in Parkinson's disease: a non-task warning paradigm study. AB - PURPOSE: the aim of this experimental study was to evaluate the attention modulating actions on the polysynaptic component of blink reflex responses and especially of the R3 component in patients suffering from Parkinson's Disease (PD). To this end, a non-task warning paradigm was adopted. METHODS: attention processing was evaluated by means of a non-task paradigm in 55 patients suffering from PD. Subjects were presented with a visual 'warning' prestimulus and the blink reflex (BR) analyzed with special regard for any modulation of its polysynaptic components (R2-R3). RESULTS: The mean amplitude of the post-warning R3 component (PW-R3c) of 'de novo' PD patients was 62% of the corresponding component following unannounced stimuli, a figure which differs significantly from both treated PD patients (18.9%) and control subjects (15.4%). De novo patients subsequently started on L-dopa therapy exhibited a more pronounced inhibition of the R3 component after warning stimulus, as the PW-R3c percentage decreased. Inversely, treated patients whose therapy was withheld showed decreased inhibition of this component. Regarding R2, the mean PW-R2c in the de novo patients differed slightly from that of the treated patients (P<0.05), but not from that of the control subjects. Such a finding may be attributable to a specific effects on the excitability of the polysynaptic responses. CONCLUSIONS: Attention disorders in PD have been well documented by means of this kind of non task warning paradigm, which appears to probe the modulation of the BR R3 component, even if the interpretation of this R3 changes suggesting a specific alteration of attention processing must be put forward extremely carefully, because something similar, but less evident, appears also for R2. PMID- 11852160 TI - Choosing an equivalence limit for noninferiority or equivalence studies. AB - Studies that compare treatments with the purpose of demonstrating that the treatments are similar require an a priori definition of an equivalence limit, how different the treatments can be before the difference is of concern. Defining such an equivalence limit is one of the most difficult aspects of planning the study. Three principles are proposed for setting such limits, depending on the objective of the study: a putative placebo calculation, an approach based on clinically important differences, and methods based on statistical properties. All methods will be useful for many studies, but the study objective should determine the final choice of an equivalence limit. The statistician must play an integral role in determining the final equivalence limit. Advice is offered for helping the statistician participate in the decision on the equivalence limits. PMID- 11852161 TI - Utility and pitfalls of some statistical methods in active controlled clinical trials. AB - Increasingly often, the study objective in an active controlled clinical trial without a placebo arm is to show that a new treatment is no less effective than the active control treatment within some noninferiority range. Two issues behind this objective are that of whether the new treatment is efficacious relative to a putative placebo and that of whether the new treatment preserves a certain fraction of effect of the active control. To address these issues, two types of statistical analysis methods are employed in recent pharmaceutical applications. In one type of method, a noninferiority margin is determined, and then the relative effect of the new treatment versus the control is compared against the margin to test noninferiority and the efficacy of the new treatment. In the other type of method, a synthetic statistic is constructed to directly estimate or test the effect of the new treatment relative to the putative placebo without resorting to noninferiority argument. Preservation of control effect can also be estimated and tested. These methods carry some crucial assumptions. The effect of active control is often estimated from a collection of historical placebo controlled trials using the random effects modeling of DerSimonian and Laird. In this work we find that statistical validity of the latter method rests highly on the assumptions that control effect is not reduced in the current active controlled trial population compared to the historical trials and that a normal approximation is appropriate in the random effects modeling. This type of method is very sensitive to departure from these assumptions. In contrast, the former method is ultraconservative in terms of type I error when the assumptions are met and can be anticonservative when control effect is substantially less in the active controlled trial than estimated from the historical placebo controlled trials. PMID- 11852162 TI - The quality of conduct in Japanese clinical trials: deficiencies found in GCP inspections. AB - The quality of commercially sponsored clinical studies in Japan was examined with reference to deficiencies in the audit reports issued by the Organization for Pharmaceutical Safety and Research (OPSR). The OPSR is responsible for domestic good clinical practice auditing in Japan. Routine audits from 1997 to 2000 for 331 hospitals revealed various types of deficiencies such as errors in case report form (CRF) entries, institutional review board problems, and protocol deviations. The high prevalence of CRF-related deficiencies seemed to stem from peculiarities of the Japanese study environment such as the historical lack of on site monitoring by the sponsor and the absence of research nurses. Characteristics in usual medical practices such as multiple drug use and loose informed consent also seemed to be associated with prevalence of similar deficiencies in clinical trials. PMID- 11852163 TI - The quality and characteristics of clinical drug study notifications reviewed by the regulatory agency in Finland. AB - The aim of our study was to investigate the validity of clinical drug study notifications reviewed by the regulatory agency in Finland during the 1990s. (In practice, the notification is equivalent to tacit authorization, which the agency has full powers to revoke before it takes effect.) All clinical drug studies reviewed by the agency during the years 1992, 1994, 1996, and 1998 were studied retrospectively. The main measurements used were the number of studies with no objection to start; the number and type of questions raised; the profile, phase, and type of study; and the study design. Additionally, the studies approved by two ethics committees of university hospitals during the same years were cross checked to see whether the agency was notified of them in accordance with the national regulations. In total, 1174 study notifications were reviewed. Most studies were international (52%), phase III (46%), placebo-controlled with/without active control (35%) investigations of new chemical entities (38%) and were carried out in university hospitals (63%). The regulatory agency had no objections or questions regarding 55% of the notifications; 37% of the studies were permitted to begin after a clarification; 5% had to be clarified a second time; and 3% were rejected. Most questions dealt with subject information. Out of the 1140 permitted studies, 8% were later canceled or prematurely terminated as reported by the applicant. Altogether 71% of the studies that had been reviewed and approved by the ethics committees were reported to the authorities before commencement. Study completions were rarely reported. Most of the clinical drug studies planned in Finland are large international studies to investigate new chemical entities. More than half of the notifications are valid according to the regulatory authorities. Not all studies, nor the majority of study completions, are reported to the authority, though according to the regulations they should be so reported. The results show that better compliance with regulatory requirements is needed, and the contents of submitted documents should be improved to gain better Good Clinical Practice compliance. The regulatory agencies and committees that review clinical study documents should improve their current practices by a more specific division of responsibilities. PMID- 11852164 TI - Showing a treatment is good because it is not bad: when does "noninferiority" imply effectiveness? PMID- 11852165 TI - The issues to be considered in global drug development. PMID- 11852166 TI - Regulatory approvals in a large multinational clinical trial: the ESPRIT experience. AB - While accepted as serving an important function to safeguard human subjects, the process of obtaining regulatory approvals to conduct clinical trials is generally regarded as cumbersome and time-consuming. For large multinational trials, U.S. federally sponsored human subject research abroad involves specific U.S. regulatory requirements, in addition to those of the host country, that act as further hurdles. These requirements may include obtaining an Assurance of Protection for Human Subjects from the Office of Human Research Protection of the U.S. Department of Health and Human Services, maintaining specific Ethics Committee/Institutional Review Board (EC/IRB) composition, and incorporating mandated elements in informed consents, all of which may differ from local policies and guidelines. Specific examples of issues that led to delays in regulatory approvals for sites participating in the multinational clinical trial entitled Evaluation of Subcutaneous Proleukin in a Randomized International Trial (ESPRIT) are presented here. While the goal of these requirements is to protect the rights and welfare of human subjects, they may create substantial delays and engender resentment over the notion of lack of respect for individual country sovereignty. Substudies within ESPRIT have been undertaken to obtain feedback from EC/IRB chairpersons, site personnel responsible for processing the required assurances, ESPRIT investigators, and study participants regarding aspects of current U.S. regulatory requirements related to human subject protection and ethical issues in multinational research. The purpose of these substudies is to compare the attitudes and experiences across countries regarding important ethical issues associated with conducting ESPRIT. One objective of the substudies is to gather additional insight to the impact of U.S. regulatory processes. Another is to help to inform the debate about how to best maximize the rights and welfare of clinical trial participants without delaying the initiation of research, while respecting the importance of national sensitivities. PMID- 11852167 TI - Crossing international boundaries: implications for the Term Breech Trial Data Coordinating Centre. AB - The Term Breech Trial (TBT) is a multicenter, international randomized trial that compared a policy of planned cesarean section with a policy of planned vaginal birth for selected pregnancies with a fetus in breech presentation at term. The TBT involved 121 centers in 26 countries that recruited 2088 women between January 9, 1997 and April 21, 2000. This paper briefly describes the impact of broad international collaboration on data coordinating center activities, including center selection, obtaining ethics approvals, data management, center funding, and recruitment. PMID- 11852168 TI - Implementing the EGASIS trial, an international multicenter acute intervention trial in stroke. AB - This paper describes some of the problems that were encountered during implementation of the Early GABA-ergic Activation Study In Stroke (EGASIS), a large international multicenter clinical trial evaluating the neuroprotective effect of diazepam in acute stroke. The focus is on obtaining funding for such a large international nonprofit trial, getting approval from all national and local ethics committees, shipping trial medication, and trial insurance. For each topic the specific problems and ways in which they were solved are described. Several recommendations for facilitating the running of a large international multicenter clinical trial such as EGASIS are made. PMID- 11852169 TI - Streamlining IRB review in multisite trials through single-study IRB Cooperative Agreements: experience of the Beta-Carotene and Retinol Efficacy Trial (CARET). AB - With their extensive data and specimen repositories, clinical trials are a long term, valuable resource to health researchers. However, assuring protection of participants' rights can be challenging, particularly when such trials are conducted at multiple sites with multiple Institutional Review Boards (IRBs). One little-used mechanism that can streamline IRB review in multisite trials while maintaining participants' protections is the single-study IRB Cooperative Agreement. This agreement is entirely different from reciprocity agreements between institutions. Beginning in 1996, the Beta-Carotene and Retinol Efficacy Trial established single-study IRB Cooperative Agreements among its performance sites, which reduced the average time to complete IRB approval from over 6 months to 1 month for each of many substudies. We describe our experience and make recommendations for other multisite clinical trials. PMID- 11852170 TI - Drug distribution for a large crossover trial of the safety of inactivated influenza vaccine in asthmatics. AB - We report on the drug packaging, masking, and distribution procedures for a large double-masked, randomized, controlled, crossover trial. The Study of Inactivated Influenza Vaccine in Asthmatics was performed to evaluate whether influenza vaccination was safe for asthmatics. Nineteen clinical centers enrolled 2032 participants from 15 September to 30 November 2000. The participants were assigned individual kits with two syringes (placebo and vaccine); the order of injection was randomly assigned. All participants enrolled in the trial received the first injection and 2009 participants received the second injection. Five participants received their injections in the wrong order; we have no evidence that any participant received an injection from the wrong kit. Challenges faced by the trial were a manufacturing shortage of vaccine, the lack of a commercially available placebo, short time periods for planning and startup, and a diverse set of clinical centers. Utilizing individual syringe kits, using the same identification for patient, kit, and drug assignment, and dispensing drug from a central facility helped to successfully meet these challenges. PMID- 11852171 TI - Double placebo design in a prevention trial for Alzheimer's disease. AB - The Alzheimer's Disease Anti-inflammatory Prevention Trial (ADAPT) is designed to compare two nonsteroidal anti-inflammatory agents against placebo. A placebo control for two active treatments can be achieved in several ways, each of which may have different implications for the logistics of drug supply, enrollment, adherence to treatment, and the validity of specific treatment comparisons. The authors outline the placebo designs considered in ADAPT, discuss their advantages and disadvantages, and provide their rationale for and the consequences of the selection of the double placebo design. PMID- 11852172 TI - Simple and effective confidence intervals for the number needed to treat. PMID- 11852175 TI - The protective effect of dl-tetrahydropalmatine against the development of amygdala kindling seizures in rats. AB - The influence of dl-tetrahydropalmatine (THP), an active component isolated from a Chinese herbal medicine corydalis, was tested on the development of electrically kindled amygdala. The seizure activity was quantified by a ultrasonic system for vertical motion measurements. Intraperitoneal injection of THP (20 or 30 mg kg(-1)) 30 min before applying the daily kindling stimulus prevented the development of the kindling process. The behavioral seizure score and the motion responses which normally develop during electrical kindling were reduced below their initial values. The results suggest that THP is a very effective antiepileptogenic and anticonvulsant agent when applied to electrically kindled rats. PMID- 11852174 TI - Aniracetam enhances glutamatergic transmission in the prefrontal cortex of stroke prone spontaneously hypertensive rats. AB - The effects of aniracetam, a cognition enhancer, on extracellular levels of glutamate (Glu), gamma-aminobutyric acid (GABA) and nitric oxide metabolites (NOx) were examined in the prefrontal cortex (PFC) and the basolateral amygdala (AMG) in stroke-prone spontaneously hypertensive rats (SHRSP) using in vivo microdialysis. Basal release of Glu, was lower in the AMG of SHRSP than in normotensive Wistar Kyoto rats, whereas no difference in GABA and NOx was noted. Aniracetam (100 mg/kg, p.o.) significantly increased the area under the curve of Glu levels in the PFC, but not in the AMG, of SHRSP. Aniracetam failed to exert any remarkable effects on GABA or NOx levels in either brain region. Our findings suggest that aniracetam enhances cortical glutamatergic release, which may be the mechanism involved in the ameliorating effects of aniracetam on various neuronal dysfunctions. PMID- 11852176 TI - Injection of substance P (SP) N-terminal fragment SP(1-7) into the ventral tegmental area modulates the levels of nucleus accumbens dopamine and dihydroxyphenylacetic acid in male rats during morphine withdrawal. AB - The biologically active substance P (SP) N-terminal metabolite SP(1-7) has been reported to modulate several neural processes such as learning, locomotor activity and reaction to opioid withdrawal. Although all these processes are believed to be associated with dopaminergic transmission no evidence of an interaction between SP(1-7) and dopamine in the case of morphine withdrawal has so far been reported. Therefore, in this work we applied in vivo microdialysis to investigate the effect of SP(1-7) injection into the ventral tegmental area on dopamine release in nucleus accumbens of male rats during naloxone precipitated morphine withdrawal. The result showed that the heptapeptide enhances dopamine release and also elevates the level of the dopamine metabolite dihydroxyphenylacetic acid in this brain area. It was suggested that the observed action of the SP fragment on the dopamine system represents the underlying mechanism for a previously observed ability of SP(1-7) to counteract the aversion response to morphine withdrawal. PMID- 11852177 TI - Adenosine 5'-triphosphate inhibits slow depolarization induced by repetitive dorsal root stimulation via P2Y purinoceptors in substantia gelatinosa neurons of the adult rat spinal cord slices with the dorsal root attached. AB - We previously reported that slow depolarization of substantia gelatinosa neurons is evoked by repetitive stimulation of C-fibers of dorsal root in adult rat spinal cord transverse slices with the dorsal root attached, which was considered to be an event relevant to spinal nociception. In the present study, we investigated the effects of adenosine 5'-triphosphate (ATP) and its analogs on the slow depolarization. ATP (10-100 microM) significantly inhibited the amplitude of slow depolarization in a concentration-dependent manner. The inhibitory effect of ATP was not reversed by suramin, an antagonist for some P2 purinoceptors, and was mimicked by a P2Y selective agonist uridine 5' triphosphate, but not a P2X selective agonist alpha,beta-methylene ATP. These results suggest that ATP inhibits the slow depolarization of substantia gelatinosa neurons relevant to nociceptive transmission in the spinal dorsal horn, via suramin-insensitive P2Y purinoceptors. PMID- 11852178 TI - Differential electrophysiological effects of brain-derived neurotrophic factor on dorsal horn neurons following chronic spinal cord hemisection injury in the rat. AB - To assess the role of brain-derived neurotrophic factor (BDNF) in nociceptive processing after chronic lateral spinal cord hemisection injury (SCI) at T13, we studied the effects of BDNF on evoked activity of dorsal horn wide dynamic range (WDR) neurons. Evoked responses of WDR cells (n=34 total) at L3-L5 were characterized electrophysiologically after spinal administration of vehicle, or BDNF (10 microg). In hemisected animals, application of BDNF to the surface of the cord resulted in reductions in evoked activity in 24 of 32 cells (75%), and enhancement of evoked activity in eight of 32 (25%) cells. Phosphate-buffered saline-receiving animals demonstrated evoked response rates of between 75 and 93 Hz, while BDNF(-) cells had evoked rates from between 20 and 41 Hz, and BDNF(+) activities were between 80 and 119 Hz, significant changes of 76 and 124%, respectively. Effects were bilateral and differences in sidedness were not observed. These results further implicate BDNF in nociceptive processing, but suggest a complex role after chronic SCI. PMID- 11852179 TI - N-methyl-D-aspartate-induced long-term depression is associated with a decrease in postsynaptic protein kinase C substrate phosphorylation in rat hippocampal slices. AB - Incubation of rat hippocampal slices with a low concentration of N-methyl-D aspartate (NMDA; 20 microM; 3 min) elicits a form of long-term depression (LTD). We used this chemical protocol to study the involvement of pre- and postsynaptic protein kinase/phosphatase activity in NMDA receptor-dependent LTD. We determined the phosphorylation states of a pre- and a postsynaptic protein kinase C substrate, B-50/growth-associated protein 43 (GAP43) and RC3, respectively, using quantitative immunoprecipitation. NMDA incubation resulted in a 2-amino-5 phosphonovalerate-sensitive long-lasting (>60 min) decrease in synaptic efficacy and a concomitant reduction in RC3 phosphorylation. B-50/GAP43 phosphorylation was unaffected. This suggests that NMDA-LTD, in contrast to low frequency-LTD, is only associated with activation of postsynaptic protein phosphatases. PMID- 11852180 TI - Histamine excites GABAergic cells in the rat substantia nigra and ventral tegmental area in vitro. AB - We have investigated the effect of histamine (HA) on spontaneous firing of dopaminergic (DA) and GABAergic neurons in the substantia nigra (SN) and the ventral tegmental area (VTA) of the rat in vitro. Single-unit extracellular recordings were obtained and drugs were bath applied. In both regions application of HA (10 and 100 microM) did not affect the firing frequency of DAergic cells, but increased the firing of GABAergic neurons. The histamine-induced excitation was blocked by the H(1) receptor antagonist mepyramine (1 microM), but was unaffected by application of the H(2) antagonist cimetidine (50 microM) or the H(3) antagonist thioperamide (10 microM). Our results suggest that histamine does not directly inhibit dopaminergic neurons in SN and VTA, but rather that this inhibition is mediated through histamine-induced excitation of GABAergic neurons. PMID- 11852181 TI - Identification of a novel transcript of X25, the human gene involved in Friedreich ataxia. AB - Friedreich ataxia (FRDA) is caused by a GAA triplet expansion in the first intron of the X25 gene. The X25 gene encodes a 210-amino acid protein, frataxin (A isoform). Here, we report the identification of a new transcript of the X25 gene generated by alternative splicing by the use of a second donor splice site in the intron 4. Full-length cDNA transcript sequence revealed an insertion of 8 bp between 4 and 5a exon sequence. This event leads to a frameshift in the mRNA reading frame and introduces a new stop codon at position 589. Therefore, this X25 transcript variant may encode a 196-amino acid protein, the A1 isoform, that structurally differs from the main A isoform of 210 amino acids after residue 160. In all human tissues analyzed, reverse transcription-polymerase chain reaction experiments demonstrated that the A1 isoform was expressed at low levels compared with the predominant A isoform. No difference in A and A1 isoform expression rate was detected between FRDA patients and normal controls. We did not find an A1 like splice variant in rodents. PMID- 11852182 TI - Quantification of mouse glial cell-line derived neurotrophic factor family receptor alpha 2 alternatively spliced isoforms by real time detection PCR using SYBR Green I. AB - Neurturin (NTN) belongs to the glial cell-line derived neurotrophic factor (GDNF) family of growth factors. Both NTN and GDNF have been shown to potently prevent the degeneration of dopaminergic neuron in vitro and in vivo. The GDNF family receptor alpha 2 (GFR alpha-2) is the preferred receptor for NTN. In addition to the known full-length isoform (GFR alpha-2a), we have previously reported the isolation of two novel alternatively spliced isoforms (GFR alpha-2b and GFR alpha 2c). All three isoforms are expressed in all mammalian tissues examined, including human fetal brain. However, the expression levels of these isoforms have yet to be quantified. In this report, we have developed a real time polymerase chain reaction (PCR) detection method using SYBR Green I to detect the expression levels of the three splice variants (GFR alpha-2a, GFR alpha-2b and GFR alpha-2c). Of the three isoforms, GFR alpha-2a was found to be the most abundant receptor expressed in the whole murine brain. The real time PCR detection method using SYBR Green I developed in this report can be used to unambiguously quantitate expression levels of the GFR alpha-2 isoforms and can be extended to the quantitation of other alternatively spliced isoforms. PMID- 11852183 TI - Human alpha-synuclein over-expression increases intracellular reactive oxygen species levels and susceptibility to dopamine. AB - alpha-Synuclein is a major component of Lewy bodies found in the brains of patients with Parkinson's disease (PD). Two point mutations in alpha-synuclein (A53T and A30P) are identified in few families with dominantly inherited PD. Yet the mechanism by which this protein is involved in nigral cell death remains poorly understood. Mounting evidence suggests the importance of oxidative stress in the pathogenesis of PD. Here we investigated the effects of wild-type and two mutant forms of alpha-synuclein on intracellular reactive oxygen species (ROS) levels using clonal SH-SY5Y cells engineered to over-express these proteins. All three cell lines, and particularly mutant alpha-synuclein-expressing cells, had increased ROS levels relative to control LacZ-engineered cells. In addition, cell viability was significantly curtailed following the exposure of all three alpha synuclein-engineered cells to dopamine, but more so with mutant alpha-synuclein. These results suggest that over-expression of alpha-synuclein, and especially its mutant forms, exaggerates the vulnerability of neurons to dopamine-induced cell death through excess intracellular ROS generation. Thus, these findings provide a link between mutations or over-expression of alpha-synuclein and apoptosis of dopaminergic neurons by lowering the threshold of these cells to oxidative damage. PMID- 11852184 TI - Developmental regulation of brain nitric oxide synthase expression in the ferret thalamic reticular nucleus. AB - We have found that cells in the ferret thalamic reticular nucleus (TRN) express brain nitric oxide synthase (bNOS) in a transient pattern during early postnatal development. Similar to our previous findings in the lateral geniculate nucleus (LGN), bNOS expression in the TRN is first observed at postnatal day 7 (P7) and continues to P35. Quantitative measures show a significant change in the relative numbers of bNOS+ cells from P7-P35, and suggest there is a transition in morphology from a bipolar shape with two primary dendrites, to a more complex, multipolar arrangement. During TRN development, the pattern of bNOS expression shifts from the somatodendritic localization seen during the first postnatal month to expression within axon fibers in the adult. Expression of bNOS within TRN cells demonstrates an additional source of nitric oxide in the developing visual thalamus, perhaps indicating a common function for thalamic nitergic neurons as cellular mediators in the establishment of central topography both in the LGN and the TRN. PMID- 11852186 TI - Repetitive spreading depression induces nestin protein expression in the cortex of rats and mice. Is this upregulation initiated by N-methyl-D-aspartate receptors? AB - In the November issue (2001) of Neuroscience Letters, Holmin et al. (Neurosci. Lett. 314 (2001) 151) reported that the synthesis of the intermediate filament protein nestin was upregulated by potassium-induced depolarization in the rat cortex. In this letter, we provide supplementary evidence that repeated cortical spreading depression elicited by potassium induces a delayed upregulation of nestin. However, we argue against the authors' conclusion, "Nestin expression was N-methyl-D-aspartate (NMDA)-receptor dependent since dizocilpine (MK-801) treatment abolished the response" because spreading depression itself is very sensitive to NMDA-receptor block, and the drug treatment was initiated prior to potassium application to the cortex in Holmin et al.'s study. PMID- 11852185 TI - Elevated levels of phosphorylated neurofilament proteins in cerebrospinal fluid of Alzheimer disease patients. AB - Neurofilament (NF) subunits NF-H, NF-M and NF-L are hyperphosphorylated and elevated in Alzheimer disease (AD) brain. We investigated the level and phosphorylation states of NF subunits in lumbar cerebrospinal fluid (CSF) from living patients by bienzyme substrate-recycle enzyme-linked immunosorbent assay. We found: (i), that the levels of phosphorylated NF-H/M (pNF-H/M), non phosphorylated NF-H/M (npNF-H/M) and NF-L were significantly higher (pNF-H/M, approximately 12-24-fold; npNF-H/M, approximately 3-4-fold) in neurologically healthy aged people than young control individuals; (ii), that in AD, the levels of npNF-H/M, and NF-L were similar to vascular dementia (VaD), and higher than in age-matched controls; and (iii), that the levels of pNF-H/M were significantly higher than in aged controls, non-AD neurological disorders and VaD. Based on these findings, it is suggested that the increased level of total NF proteins in CSF could be used as a marker for brain aging and neurodegenerative disorders in general, and the levels of pNF-H/M as a marker to discriminate AD from normal brain aging and as well as neurological conditions including VaD. PMID- 11852187 TI - Analysis of regulatory functions for the region located upstream from the latency associated transcript (LAT) promoter of pseudorabies virus in cultured cells. AB - The latency-associated transcript (LAT) promoter of pseudorabies virus (PrV) is unique among the many promoters of the viral genome in that it remains active during the latent state. The regulatory mechanism of PrV LAT gene expression is complex and different between latency and lytic infection of cultured cells. Although two different sequences, LAP1 and LAP2, are thought to be involved in LAT gene expression, the function of the upstream region of the LAT promoter (LAP1 and LAP2) remains an enigma, even in cultured cells. To analyze the function of the upstream region, it is necessary to examine the effects of the upstream sequence on LAT gene expression in the absence of other viral proteins. Transient expression assays were performed by employing a series of reporter plasmids in which various sequences upstream of the LAT promoter (from nucleotide positions -592 to +423 relative to the transcriptional start site of the large latency transcript (LLT)) were linked to the chloramphenicol acetyltransferase (CAT) gene in cells of neuronal and non-neuronal origin. We identified a region (from nucleotide positions -3606 to -1386) that was capable of repressing the LAT promoter activity in Vero cells by analyzing CAT gene expression of the series of reporter plasmids. This effect was not observed in Neuro-2a cells. We have also shown that the LAT promoter activity of the reporter plasmid containing the upstream region was repressed by the immediate-early gene product IE180 in Vero cells, but not in Neuro-2a cells. These results suggest that the upstream region of the LAT promoter may have a role in repressing LAT gene expression in cultured non-neuronal cells. PMID- 11852188 TI - Intraspecific variation in the 16S rRNA gene sequences of Mycoplasma agalactiae and Mycoplasma bovis strains. AB - Intraspecific variation in the 16S rRNA genes of 17 Mycoplasma agalactiae and eight Mycoplasma bovis isolates was investigated to determine the degree of sequence variation in these two species and to determine whether the polymorphisms in the 16S rRNA genes could be used for the construction of an evolutionary tree and as epidemiological markers. A high degree of variation was found within isolates (between operons) and between isolates of both species. In contrast to M. capripneumoniae no distinct evolutionary pattern could be seen, probably because there are functional systems for gene conversion in M. agalactiae and M. bovis. However, the non-European isolates of M. agalactiae shared three characteristic nucleotides and European isolates from the same or neighbouring countries were very similar. Differences within isolates included both polymorphic positions and sequence length differences between operons. The amount of variation within isolates of the respective species ranged from zero to seven polymorphisms for M. agalactiae and from zero to four polymorphisms for M. bovis. The high degree of variation suggests the potential for misdiagnosis of species in diagnostic PCR assays based on the 16S rRNA gene sequences. All isolates of both species had a thymidine in position 912 (E. coli numbering) that causes streptomycin resistance in several bacterial species and which is characteristic for the members of the hominis group. As expected, when five M. agalactiae and three M. bovis isolates were tested for streptomycin susceptibility, they all demonstrated streptomycin resistance. M. agalactiae and M. bovis were found to have high intraspecific variation in their 16S rRNA gene and the polymorphisms patterns indicate that gene conversion takes place. PMID- 11852189 TI - Genomic diversity among Danish field strains of Mycoplasma hyosynoviae assessed by amplified fragment length polymorphism analysis. AB - Genomic diversity among strains of Mycoplasma hyosynoviae isolated in Denmark was assessed by using amplified fragment length polymorphism (AFLP) analysis. Ninety six strains, obtained from different specimens and geographical locations during 30 years and the type strain of M. hyosynoviae S16(T) were concurrently examined for variance in BglII-MfeI and EcoRI-Csp6I-A AFLP markers. A total of 56 different genomic fingerprints having an overall similarity between 77 and 96% were detected. No correlation between AFLP variability and period of isolation or anatomical site of isolation could be demonstrated. An examination of the clonal appearance of M. hyosynoviae isolates recovered from seven herds affected with arthritis revealed presence of several genotypically distinct variants of the organism in a single herd, indicating that different strains may simultaneously be involved in an outbreak of the disease. PMID- 11852190 TI - Abomasitis associated with multiple antibiotic resistant Salmonella enterica serotype Typhimurium phagetype DT104. AB - Salmonella enterica serotype Typhimurium phagetype DT104 is a multiple antibiotic resistant pathogen that has been purported to be more pathogenic than other Salmonella. In this study, we evaluated the possibility that DT104 is the causative agent of veal calf abomasitis observed in four independent outbreaks of salmonellosis. This study was undertaken to determine if the outbreaks might be due to hypervirulent S. enterica serotype Typhimurium phagetype DT104 (DT104) since Salmonella does not usually cause abomasitis. Tissues and fluids from these calves were subjected to bacteriologic culture. Pure Salmonella cultures were then used in bovine challenge experiments. DT104 was identified as the causative agent of abomasitis in calves. Thus, abomasitis is a potential indicator of infection with multiple antibiotic resistant DT104 and adds credence to the apparent hypervirulence of this pathogen. PMID- 11852191 TI - Effect of preculture freezing and incubation on bacteriological isolation from subclinical mastitis samples. AB - In this study the sensitivity of three methods of isolation of udder pathogens from milk samples from subclinical mastitis cases was compared. For analysis 1827 quarter milk samples were selected. Milk was cultured using a standard culture technique (0.01 ml of fresh milk streaked on a sheep blood agar plate and on Edward's medium). In addition, an inoculum of 0.01 ml of the original milk sample was incubated for 24h at 37 degrees C in broth, followed by culture using the standard culture technique. In the third method, the whole milk sample was frozen for 24h, and then incubated for 24h at 37 degrees C, followed by culture using standard culture technique. The isolation percentage of Staphylococcus aureus was 4.7% for standard culture technique, 14.2% for incubation in broth, and 21.5% for the combination of freezing plus incubation. Isolation percentage of Streptococcus dysgalactiae and Streptococcus agalactiae was highest using the standard culture technique, while isolation rate of Streptococcus uberis was not different among the three methods used. With increasing somatic cell count, the likelihood of S. aureus, S. dysgalactiae and S. uberis isolation increased. Based on the relative sensitivity, defined as the isolation rate using a single technique compared to the isolation rate of the three techniques together, a combination of standard culture technique and freezing plus incubation was most attractive for achieving a high isolation rate of S. agalactiae and S. dysgalactiae. Relative sensitivity of S. uberis isolation was highest using the standard culture technique and incubation in broth, while S. aureus was most often isolated using a combination of incubation in broth and freezing plus incubation. A combination of the three methods increased the isolation rate for S. dysgalactiae, S. uberis and S. aureus. The standard culture technique, together with the combination of freezing plus incubation, can be recommended for isolating major udder pathogens. If S. aureus is the pathogen of main interest, using incubation in broth together with the combination of freezing plus incubation performed best. PMID- 11852192 TI - Development of a sandwich ELISA and comparison with PCR for the detection of F11 and F165 fimbriated Escherichia coli isolates from septicaemic disease in farm animals. AB - The P fimbriae F11 and F165 that have been demonstrated on Escherichia coli septicaemic strains in poultry and calves, respectively, possess a nearly identical major subunit that demonstrates a serological cross-reaction. A polyclonal antibody-based sandwich ELISA (sELISA) that was specific for both F11 and F165 fimbriated strains was compared with a PCR method to detect F11/F165 fimbriated strains, in a collection of E. coli strains isolated from diseased animals. Of 298 isolates tested, 36 were positive by PCR of which only 14 were sELISA positive. There were no sELISA positive but PCR negative results. The 36 PCR positive isolates comprised 11 avian strains of which 10 were sELISA positive, 20 bovine strains of which 4 were sELISA positive and 3 ovine strains, 1 porcine strain and 1 equine strain all of which were sELISA negative. The F11/F165 incidence of 10.7% in 103 poultry and 18.3% in 109 bovine isolates demonstrates a moderate level of these factors in E. coli septicaemic cases in Northern Ireland. PMID- 11852193 TI - Low molecular weight proteins of Cowdria ruminantium (Welgevonden isolate) induce bovine CD4+-enriched T-cells to proliferate and produce interferon-gamma. AB - An important objective in vaccination strategies is to activate lymphocytes with particular effector functions. Cellular immunity and the type I cytokine IFN gamma have been implicated in protective immunity to heartwater. Furthermore, low molecular weight proteins of Cowdria ruminantium have been shown to induce peripheral blood mononuclear cells to proliferate. To determine which lymphocyte subset responds when stimulated with fractionated C. ruminantium proteins, specific short-term lymphocyte cultures were established from cattle immunized with the Welgevonden isolate. Four cattle were immunized, two by infection and treatment and two with inactivated organisms. Cell surface phenotypic analysis of the cultures indicated that CD4+ lymphocytes were enriched over time. This coincided with increased antigen-specific proliferation and IFN-gamma production. Proteins of molecular weights 13-18kDa induced the CD4+-enriched T-cell cultures, derived from each of the animals, to proliferate and produce IFN-gamma. Although the two groups of cattle were immunized differently, their lymphocytes responded similarly. These results extend previous findings by identifying the responder cells as being predominantly IFN-gamma producing CD4+ lymphocytes. This cytokine has been implicated in immunity to the parasite. The low molecular weight proteins that induced CD4+ lymphocytes to proliferate and produce IFN-gamma are therefore likely to be important in protection against heartwater and may have a role in vaccine development. PMID- 11852194 TI - Replicon typing of F18 fimbriae encoding plasmids of enterotoxigenic and verotoxigenic Escherichia coli strains from porcine postweaning diarrhoea and oedema disease. AB - The presence of fimbrial adhesin F18 is frequently found in enterotoxigenic Escherichia coli (ETEC) and verotoxigenic E. coli (VTEC) strains responsible for diarrhoea and oedema disease of weaned pigs. The F18 adhesin occurs in two antigenic variants: F18ab is characteristic of VTEC while F18ac is more typical for ETEC. F18 encoding plasmids of 17 phenotypically characterized porcine E. coli isolates (10 ETEC, 6 VTEC and 1 ETEC/VTEC) were tested with a DNA probe for F18 fimbrial adhesin and with replicon probes for the RepFIa, RepFIb and for the RepFIc family of basic replicons. In all the cases, the F18 probe hybridized to only one plasmid band of size higher than 42MDa. All F18 plasmids were determined to be unireplicon plasmids belonging to the RepFIc replicon family of the F incompatibility complex. There was no difference between F18ac plasmids of ETEC and F18ab plasmids of VTEC strains in terms of replicon type or subtype. However, the size of F18ab plasmids of the VTEC strains varied between 42 and 98MDa, in contrast to F18ac plasmids of ETEC strains (constantly approximately 98MDa). PMID- 11852195 TI - Neospora caninum and Waddlia chondrophila strain 2032/99 in a septic stillborn calf. AB - Characteristics of an intracellularly growing micro-organism isolated from an aborted bovine foetus are described. The organism replicated within cytoplasmic vacuoles, was resistant to penicillin and exhibited structural characteristics compatible with Waddlia chondrophila. An ELISA specific for Chlamydia spp., immunofluorescence tests using antibodies directed against Chlamydia spp. or Simkania negevensis, and PCR using Chlamydia-specific primers showed that the agent was distinct from Chlamydiae or S. negevensis. Determination of 16S and partial 23S ribosomal RNA gene sequences in combination with the PCR results and the morphological, antigenic and developmental characteristics provided evidence that the isolate 2032/99 can be classified as W. chondrophila or a closely related organism. PMID- 11852198 TI - Blind field test evaluation of Raman spectroscopy as a forensic tool. AB - Analytical instrumentation for Raman spectroscopy has advanced rapidly in recent years to the point where commercial field-portable instruments are available. Raman analysis with portable instrumentation is a new capability that can provide emergency response teams with on-site evaluation of hazardous materials. Before Raman analysis is accepted and implemented in the field, realistic studies applied to unknown samples need to be performed to define the reliability of this technique. Studies described herein provide a rigorous blind field test that utilizes two instruments and two operators to analyze a matrix that consists of 58 unknown samples. Samples were searched against a custom hazardous materials reference library (Hazardous Material Response Unit (HMRU) Spectral Library Database). Experimental design included a number of intentionally difficult situations including binary solvent mixtures and a variety of compounds that yield medium-quality spectra that were not contained in the HMRU library. Results showed that over 97% of the samples were correctly identified with no occurrences of false positive identifications (compounds that were not in the library were never identified as library constituents). Statistical analysis indicated equivalent performance for both the operators and instruments. These results indicate a high level of performance that should extrapolate to actual field situations. Implementation of Raman techniques to emergency field situations should proceed with a corresponding level of confidence. PMID- 11852196 TI - Results of the 1999-2000 collaborative exercise and proficiency testing program on mitochondrial DNA of the GEP-ISFG: an inter-laboratory study of the observed variability in the heteroplasmy level of hair from the same donor. AB - The Spanish and Portuguese working group (GEP) of international society for forensic genetics (ISFG) 1999-2000 collaborative exercise on mitochondrial DNA (mtDNA) included the analysis of four bloodstain samples and one hair shaft sample by 19 participating laboratories from Spain, Portugal and several Latin American countries. A wide range of sequence results at position 16,093 of the HV1 (from T or C homoplasmy to different levels of heteroplasmy) were submitted by the different participating laboratories from the hair shaft sample during the first phase of this exercise. During the discussion of these results in the Annual GEP-ISFG 2000 Conference a second phase of this exercise was established with two main objectives: (i) to evaluate the incidence of the HV1 sequence heteroplasmy detected in Phase I across different sample types from the same donor including blood, saliva, and hair shafts, (ii) to perform a technical review of the electropherograms to evaluate the relative levels of heteroplasmies obtained by the different laboratories and also to examine the source of possible errors detected in Phase I. Anonymous review of the raw sequence data permitted the detection of three transcription errors and three errors due to methodological problems. Highly variable levels of heteroplasmy were found in the hair shaft and more stability in blood and saliva. Three laboratories found variable levels of heteroplasmy at position 16,093 across adjacent fragments from the same hair shaft. Two laboratories also described more than one heteroplasmic position from a single hair. The relevance of these findings for the interpretation of mtDNA data in the forensic context is also discussed. PMID- 11852200 TI - Elemental analysis of bone: proton-induced X-ray emission testing in forensic cases. AB - Proton-induced X-ray emission (PIXE) is a spectroscopic technique that provides the researcher with the elemental composition of a given target material. In this paper, we illustrate the utility of PIXE analysis in two forensic contexts: (1) case of cremation in which the nature of the remains is questioned and (2) cases of death by gunshot wound. In the first case, elemental analysis by PIXE reveals that the purported cremated remains are not bone. The last two cases show that radiopaque metallic residue embedded in bone is composed of lead from a projectile. PMID- 11852201 TI - High resolution Y chromosome typing: 19 STRs amplified in three multiplex reactions. AB - Nineteen Y-specific short tandem repeat (STR) loci have been amplified in 768 samples from the Iberian Peninsula in order to evaluate their usefulness in forensic casework. Two previously published multiplex reactions by Thomas et al. [Hum. Genet. 6 (1999) 577] (MS1, modified here: DYS19, DYS388, DYS390, DYS391, DYS392 and DYS393) and by Ayub et al. [Nucl. Acids Res. 28 (2000) e8] (CTS: DYS434, DYS435, DYS436, DYS437, DYS438 and DYS439) plus a novel one reported here (EBF: DYS385, DYS389, DYS460, DYS461, DYS462 and amelogenin) have been used. DYS385, DYS439 and DYS391 were the most informative loci with allele diversities of 0.7997, 0.6683 and 0.5940, respectively. A total of 635 different haplotypes were observed, of which 573 (90.24%) were found in single individuals. The overall haplotype diversity was 0.9988 and that obtained by each multiplex system was 0.9812 for EBF, 0.9292 for MS1 and 0.9089 for CTS. PMID- 11852202 TI - Jewish population genetic data in 20 polymorphic loci. AB - Frequency data of 20 polymorphisms (13 autosomal and 6 Y-chromosome STRs and the Alu-insertion YAP) were determined in a sample of 124 unrelated Jewish individuals from Ashkenazi, Sephardic, North African and Oriental origin. The a priori statistical power of the set of studied loci was highly discriminating (PD=1 in 2.393 E+15 individuals for autosomal STRs, and a probability of finding two unrelated males with the same haplotype of less than 0.01). Some significant population differences were registered between Jews and other Circummediterranean populations in Y-chromosome loci frequencies. PMID- 11852203 TI - Postmortem serum uric acid and creatinine levels in relation to the causes of death. AB - Serum uric acid (UA) and creatinine (Cr) mainly derive from skeletal muscle tissues. Although, remarkable postmortem stability of the serum levels has been reported, there appears to be very poor knowledge of the diagnostic value in investigation of death, except for uremia. The aim of the present study was to evaluate postmortem serum UA and Cr levels using 395 forensic autopsy cases, in comparison with blood urea nitrogen (BUN), for investigation of the pathophysiology of death with special regard to the causes of death involving possible skeletal muscle damage, e.g. due to hypoxia, heat or agonal convulsions. Cr and BUN showed relatively good topographic stability in the cadaveric blood, whereas, UA was often much higher in the right heart blood than in the left heart and peripheral blood, independent of postmortem intervals. Moderate to marked elevation of Cr and BUN accompanied with hyperuricemia was observed in delayed death. In the acute death cases (survival time <30 min), UA, especially in the right heart blood, showed a considerable elevation in mechanical asphyxiation and drowning. The Cr level in fire victims with a lower carboxyhemoglobin (COHb) level (<60%) was significantly higher than in those with the possible fatal level (>60%). A similar elevation of Cr was observed in fatalities from heat stroke and methamphetamine (MA) poisoning. The observations suggested that hyperuricemia in acute death may be indicative of advanced hypoxia and that elevated Cr level may reflect the skeletal muscle damage, especially due to thermal influence. PMID- 11852204 TI - Improved estimation of postmortem interval based on differential behaviour of vitreous potassium and hypoxantine in death by hanging. AB - Many formulae are available to estimate the relation between the potassium ([K+]) and hypoxantine ([Hx]) concentration in the vitreous humour and the postmortem interval (PMI). Typically these have been based on a correlation test and linear regression using the postmortal interval as the independent variable and [K+] or [Hx] as the dependent variable in order to estimate the confidence interval. However, a recent study has shown that a more precise measurement of PMI can be obtained if [K+] is used as the independent variable. The regression lines obtained from the most recent deceased subjects with forensic relevance received for autopsy in the Institute of Legal Medicine are [K+] = 5.589 + 0.174PMI and [Hx] = 26.459 + 3.017PMI, by changing the variables, we obtain PMI=3.967[K+] - 19.186 (R2 = 0.688, P < 0.001) and PMI = 0.172 [Hx] + 0.170 (R2 = 0.518, P < 0.001). In this paper we propose the cause of death as an extra factor which modifies the relationship and gives even greater precision in estimating PMI. In cases of death by hanging the results are considerably improved with [K+] = 5.224 + 0.225PMI and [Hx] = 15.161+4.957PMI, respectively, and consequently, PMI = 3.631[K+] - 17.334 (R2 = 0.818, P< 0.001) and PMI = 0.153[Hx] - 0.368 (R2 = 0.757, P < 0.001): the slope is less and the precision is obviously enhanced. PMID- 11852205 TI - Drugged drivers in Norway with benzodiazepine detections. AB - Norwegian drugged drivers with benzodiazepine (BZD) detections have been studied with regard to drug use pattern and rearrest rate. During 1995, 3343 drivers were apprehended by the police in Norway due to the suspicion of influence by drugs. Blood samples from all drivers were sent to the National Institute of Forensic Toxicology (NIFT). The samples were analysed using a standard program covering the most commonly abused drugs on the marked in Norway. BZDs, representing some of the most frequently detected drugs, were found in approximately 30% (n = 1051) of the cases, represented by 14% (n = 150) female and 86% (n = 901) male drivers. In 8% of the cases, one BZD only was detected, half of these cases with one BZD could reflect therapeutic use. One or more BZDs were combined with illegal drug(s) (73%), other prescribed drugs (10%), and/or alcohol (24%). 62% of the drivers with BZD detections, had earlier been arrested for the same offence, or six cases per rearrested driver. The frequency of earlier arrests were lower for female (34%) than for male (67%) drivers. Alcohol was most frequently found for those arrested for the first time before 1992, while BZD or illegal drugs were most frequently found for those with their first arrest during 1992-1995. Our study shows that apprehended drivers using BZD are mainly represented by drug abusers due to frequent multi-drug use, blood concentrations representing doses above therapeutic levels and high rearrest rate for the same offence. A treatment program or other reactions, are thus necessary in addition to fines, prison penalty and suspension of driving licence. PMID- 11852206 TI - Analysis of 13 tetrameric short tandem repeat loci in a population of Tuscany (Central Italy) performed by means of an automated infrared sequencer. AB - Allele frequencies for the 13 STRs of the Combined DNA Index System (CODIS) core were obtained from a sample of 188 unrelated individuals living in the area of Florence, Prato and Pistoia (Tuscany, Central Italy). PMID- 11852207 TI - Y-chromosome STR haplotypes in a southwest Spain population sample. AB - The Y-chromosome polymorphism of eight STRs (DYS19, DYS389I, DYS389II, DYS390, DYS391, DYS392; DYS393, DYS385) were studied in 111 unrelated individuals from the population of southwest Spain. The most common haplotype was shared by 3.6% of the sample, while 99 haplotypes were unique. The gene diversity was 0.9977. PMID- 11852208 TI - Population genetics of the STRs D3S1358, FGA, D2S1338, D8S1179, D21S11, D18S51 and D19S433 in the Pomerania-Kujawy region of Poland. AB - Allele frequencies for seven STRs were obtained from a sample of 789 unrelated individuals born in the Pomerania-Kujawy region of Poland. PMID- 11852209 TI - Mitochondrial sequence haplotype in the Japanese population. AB - Sequence polymorphysms of the mitochondrial DNA (mtDNA) control region, hypervariable regions I and II, from 50 unrelated Japanese were determined by PCR amplification and cycle sequencing. PMID- 11852210 TI - Using a bidimensional t test to compare simultaneous activations in functional brain MRI. AB - The aim of functional brain magnetic resonance imaging studies is to determine which regions of the brain are related to a given specific task. Different methods can be used to extract the functional signal and there is currently no consensus for this operation. Measures based on correlation are only relevant for a single task. In this paper, we propose a method based on a multivariable Student's t test that permits the comparison of the results of successive activations. This technique allows a qualitative analysis of complex tasks, making possible to deal with both similar and different activated cortical areas. PMID- 11852211 TI - Transcranial color-coded duplex sonography of the carotid siphon: the coronal approach. AB - Transcranial color-coded sonography can assist in the assessment of the carotid siphon segments C1 and C5 employing two standardized coronal image planes. In 32 volunteers mean velocities in the C1 and C5 were 41+/-16 and 30+/-10 (cm/s+/ S.D.), pulsatility indices (PI) 0.92+/-0.22 and 0.93+/-0.27, and resistance indices (RI) 0.58+/-0.08 and 0.55+/-0.12, respectively. Concluding, this technique is useful in assessing the carotid siphon with the most robust parameters being PI and RI requiring further validation by the "gold standard" angiography. PMID- 11852212 TI - Laryngeal amyloidosis: a rare cause of laryngocele. AB - Laryngocele may be defined as an abnormal dilatation of the saccule of the ventricle and its pathogenesis is uncertain. We report a laryngocele caused by amyloidosis localized to larynx in a 60-year-old male patient. We would like to mention that amyloidosis should be evaluated in the etiology of patients diagnosed as having a laryngocele. PMID- 11852213 TI - MR findings with dynamic evaluation in Riedel's thyroiditis. AB - We reported the findings on MRI in a patient with proven Riedel's thyroiditis. The lesion was seen as slightly heterogeneous hypointensity on both T1- and T2 weighted images and had a mild enhancement after administration of gadpentate dimeglumine (Gd-DTPA). Dynamic MR study showed gradual increase in signal intensity of the lesion. These findings were different from those of thyroid neoplasms and were considered to reflect the mixture of inflammatory cells infiltration and fibrosis of Riedel's thyroiditis. PMID- 11852214 TI - Frequency of sinusitis in the patients with pneumonia. AB - The aim of this study was to investigate the frequency of sinusitis in patients with pneumonia. Among 50 consecutive patients with pneumonia, 19 cases, which had Waters' roentgenograms in their files, were retrospectively reviewed in relation with sinusitis. Sixteen of them had sinusitis signs in their roentgenograms. The sinusitis frequency was 84% in patients with pneumonia who had paranasal sinus roentgenograms. These findings suggest that sinusitis may be an important causative factor for pneumonia and that it should be considered when evaluating a patient with pneumonia. PMID- 11852215 TI - Bronchioloalveolar carcinoma and adenocarcinoma with bronchioloalveolar features presenting as ground-glass opacities on CT. AB - OBJECTIVE: As bronchioloalveolar carcinoma (BAC) is noninvasive but, in its later stages, has a worse prognosis than adenocarcinoma with bronchioloalveolar features (ACB), early identification and differentiation is important for therapeutic and prognostic purposes. We wanted to identify features of BAC, which differentiated it from ACB when both presented as ground-glass opacities (GGOs) on CT. MATERIALS AND METHODS: We reviewed all pathologic specimens of patients who were diagnosed with BAC and ACB in the lung from 1991 to 1999 in our institution and whose malignancy presented as a GGO on CT. This yielded 29 patients, 15 with BAC and 14 with ACB with GGOs on CT. Both univariate frequency table and multivariate logistic regression approaches were used to analyze the CT characteristics of these GGOs (location, GGO pattern, size, shape, margin, presence and type of air bronchogram and pseudocavitation). RESULTS: BAC most frequently had a "GGO halo" around a solid opacity, often was a GGO "mixed with consolidation" with the smallest BACs being "pure GGO." Air bronchograms were frequently present in the largest GGOs. Pseudocavitations were rare. ACB, on the other hand, most frequently presented as a GGO "mixed with consolidation," less frequently with a "GGO halo" and rarely with "superimposed lymphangitis." The air bronchograms, frequently present, were usually tortuous and ectatic. Pseudocavitation was present in about one-third of the cases. The most useful CT features of GGO in separating those due to BAC from those due to ACB were pure (uniform) ground-glass attenuation and absence of lymphangitis. CONCLUSION: The CT features of BAC and ACB presenting as GGO reflect the histologic descriptions of these carcinomas. PMID- 11852216 TI - CT pulmonary angiography in the detection of pulmonary emboli: a meta-analysis of sensitivities and specificities. AB - OBJECTIVE: To determine the overall sensitivity and specificity for CT pulmonary angiography (CTPA) in the diagnosis of pulmonary emboli (PE) using a meta analysis of the published literature. MATERIALS AND METHODS: A Medline search was constructed to include all English language publications indexed in the Index Medicus from 1990 to 2000, which included the terms CT, PE and pulmonary angiography. Studies selected were designed principally to compare CTPA in the overall detection of PE as confirmed by an abnormal fluoroscopic pulmonary angiogram or a high probability V:Q scan. Results were corrected for the patient sample size in the respective studies prior to pooling the data. In the absence of an accepted technique for calculating a ROC curve in the meta-analysis of imaging studies, a previously untested theoretical technique was used to obtain a composite ROC curve. RESULTS: Twelve studies of CTPA comprising a total of 1250 patients were analyzed. The overall sensitivity and specificity for CTPA after correction for study size was 74.1% and 89.5% with a range of 57-100% and 68 100%, respectively, for the detection of PE. No trend was detected with respect to the year of publication or sample size. CONCLUSIONS: CTPA has acceptable sensitivity and specificity with a strong ROC curve making it a good first line investigation for PE. PMID- 11852217 TI - Influence of heart rate on vessel visibility in noninvasive coronary angiography using new multislice computed tomography: experience in 94 patients. AB - OBJECTIVES: Initial reports indicate that coronary artery lesions might be visualized with high sensitivity and specificity by the use of recently introduced multislice computed tomography (MSCT). Current CT technology offers a temporal resolution of 250 ms. In case of heart rates (HRs) >65 beats/min (bpm), however, the reconstruction software switches from a single-phase algorithm (using data from one heart cycle only) to a biphase algorithm using image data of two consecutive heart cycles, improving temporal resolution to down to 125 ms. Thus, it was the aim of the present study to evaluate the influence of the patients' (pts) HR on image quality expressed by vessel segment visibility. METHODS AND RESULTS: MSCT scans (Somatom VZ) were performed in 94 pts. Ten coronary segments were analyzed in each patient with regard to image quality (RCA: segments [sgts] 1-4, LMS: sgt 5, LAD: sgts 5-8, LCX: sgts 11, 12). A total of 697 of 940 (74.1%) sgts were accurately visualizable (RCA: 244/376 [64.9%], LMS: 94/94 [100%], LAD: 232/283 (82.3%), LCX: 146/188 [77.7%]). Beta-blocker therapy had a significant influence on mean HR (65 pts on beta-blocker, HR 65.1+/ 10.7 bpm vs. 29 pts, HR 71.6+/-12.2 bpm, P=.01). A significant inverse correlation between HR and segmental visibility was found (r=-.48, P<.0001), with best visibility in pts with lower HRs (n=14 pts with 10 analyzable sgts, HR 60+/ 10.1 vs. n=8 pts with 4 analyzable sgts, HR 79.9+/-6.9, P<.0001). CONCLUSIONS: Our results indicate that vessel visibility is highly dependent on the pts HR. Best vessel visibility was found in pts with HR <65 bpm with single-phase image reconstruction. Thus, it appears to be advisable to evaluate, and if needed, to lower the pts HRs before undergoing MSCT coronary angiography in order to achieve best image quality. PMID- 11852218 TI - CT and MRI in tuberculous sternal osteomyelitis: a case report. AB - We report a 58-year-old male patient presenting with a 1-year history of presternal swelling and pain. Plain radiography revealed increased soft tissue density anterior to the body of the sternum, which showed cortical sclerosis. Computed tomography (CT) demonstrated ring-enhancing hypodense soft tissue masses surrounding the sternum, whose anterior and posterior cortices were markedly thickened. On three-phase technetium bone scintigraphy, the left side of the sternum showed increased radiotracer uptake and the central part of the bone was photopenic. The bone marrow of the sternum and peristernal soft tissue lesions were hypo- and hyperintense on T1- and T2-weighted magnetic resonance (MR) images, respectively, and showed marked enhancement postgadolinium. Treatment included both surgical intervention and medical therapy. PMID- 11852219 TI - The "skinny aorta". AB - This article reviews the imaging findings of focal narrowing of the descending thoracic and/or abdominal aorta and stresses the clinical manifestations that may help differentiate specific entities. Various entities including coarctation of the aorta, Takayasu's arteritis, neurofibromatosis, radiation therapy, and infantile hemangioendothelioma are discussed and key differential points are emphasized. PMID- 11852220 TI - Liver volume measurement by spiral CT: an in vitro study. AB - Liver volume measurement has many clinical and research uses. The purpose of the present study is to evaluate experimentally the accuracy and reproducibility of spiral computed tomography (CT) in measuring liver volume. To determine the accuracy of spiral CT system, nine fresh (non-formalin-fixed) sheep livers were scanned in vitro. The collimation was 7 mm. The CT sections were stored on floppy disks and the images were transferred to the workstation that contained a software for volume measurement. The mean volume is 912+/-497 ml and the actual reference volume is 921+/-527 ml. The correlation between the CT system and actual reference measurement of sheep liver volumes was very strong. The correlation coefficient r was.998 (P<.001) on 7-mm sections. The mean difference between liver volume measurements was 4.95+/-3.23%. As a result, CT scan could be used as a reference noninvasive method for liver volume measurement in clinics. PMID- 11852221 TI - Giant hydronephrosis due to ureteropelvic junction obstruction in a child: CT and MR appearances. AB - Giant hydronephrosis caused by congenital ureteropelvic junction (UPJ) obstruction is a rare urological entity in childhood. We report a case of giant hydronephrosis in a 12-year-old boy presented with abdominal distension. Radiological features of this rare entity are discussed with an overview of sonographic findings, especially including CT and MR appearances. The current literature is also reviewed. PMID- 11852222 TI - Periprostatic infiltration with local anesthesia during transrectal ultrasound guided prostate biopsy is safe, simple, and effective: a pilot study. AB - OBJECTIVE: Transrectal ultrasound (TRUS)-guided prostate needle biopsy is the standard procedure to diagnose prostate cancer. It can be associated with significant discomfort and pain. We evaluated if periprostatic infiltration with local anesthetic reduces this discomfort. MATERIAL AND METHODS: 72 patients underwent TRUS-guided prostate needle biopsy. All patients had 12 cores, with four prostatic zones of biopsies. In 25 consecutive patients (G1) with a median prostate size of 47 cc, no anesthesia was given; while in 47 sequential patients (G2) with a median prostate size of 50 cc, 2 ml of 2% lidocaine was infiltrated in the periprostatic area around the neurovascular bundle using 20-cm-long, 22 gauge needle on both sides as guided by color Doppler. Biopsies were performed in standard fashion. Discomfort was graded on a scale from 0 to 10 with 0 meaning no discomfort, 1-3 mild, 4-6 moderate, and 7-10 severe. RESULTS: 12/25 (48%) of G1 patients reported no discomfort compared to 70% in G2 (P=.025). Mild discomfort was reported in 5/25 (20%) patients of G1 and 9/47 (19%) patients of G2. Moderate or severe discomfort was reported in 8/25 (32%) patients and 5/47 (11%) patients in G2 (P=.039). Prostate size did not affect degree of discomfort within each group and between both subgroups. No adverse reactions were observed secondary to lidocaine infiltration. CONCLUSION: Periprostatic infiltration with local anesthesia at the time of TRUS-guided prostate needle biopsy significantly reduces discomfort. It is easy to perform, safe, and should be considered in all patients irrespective of the prostate size. PMID- 11852223 TI - Small cell carcinoma of the uterine corpus: CT appearance. AB - We report a rare case of small-cell carcinoma of the uterine corpus. To our knowledge, this is the first radiological report. Computed tomography (CT) demonstrates a hypodense lesion within the endometrial cavity with nonhomogeneous contrast enhancement. PMID- 11852224 TI - Evaluation of the patellofemoral joint with kinematic MR fluoroscopy. AB - OBJECTIVE: The aim of this study was to evaluate the diagnostic effectiveness of kinematic magnetic resonance fluoroscopy (KMRF) on patients with patellofemoral incongruency. MATERIALA AND METHODS: 17 patients (20 knees) and 10 healthy volunteers (20 knees), all men (mean age 29.4 years, S.D. 9, range 16-50), were included to our study. Only male subjects were studied because of potential biomechanical differences between sexes. KMRF was used to perform kinematic MR imaging of patellar alignment and tracking in 10 healthy subjects and 17 patients with a provisional clinical diagnosis of abnormal patellofemoral joints. The patellofemoral joints were examined with the knee in different angles of active flexion. At each knee position, real time kinematic and axial MRF image was used to focus on the sagittal plane, followed by an axial image focused through the middle of the patella. Three angles were measured: patellar tilt angle (PTA), sulcus angle (SA) and congruence angle (CA). RESULTS: Five patterns of malalignment were identified and studied. Two patellofemoral joints were normal, 10 had lateralization of the patella, 2 had patellar tilt, 2 had lateralization and patellar tilt (i.e. excessive lateral pressure syndrome) and 4 had medialization of the patella. CONCLUSION: KMRF is an effective method in evaluating patellofemoral incongruency. Short time duration of investigation, ability to get nearly real time images, suitable temporal contrast resolution and investigation from very different angles of knee are important advantages of the method. PMID- 11852225 TI - Tarsal tunnel syndrome caused by talocalcaneal coalition. AB - Tarsal tunnel syndrome caused by talocalcaneal coalition is uncommon. We presented the ultrasonography (US) and magnetic resonance imaging findings of this disease. This is, to our knowledge, the first case report describing the US findings in tarsal tunnel syndrome caused by talocalcaneal coalition. PMID- 11852228 TI - Domain structure and organisation in extracellular matrix proteins. AB - Extracellular matrix (ECM) proteins are large modular molecules built up from a limited set of modules, or domains. The basic folds of many domains have now been determined by crystallography or NMR spectroscopy. Recent structures of domain pairs and larger tandem arrays, as well as of oligomerisation domains, have begun to reveal the principles underlying the higher order architecture of ECM proteins. Structural information, coupled with site-directed mutagenesis, has been instrumental in showing how adjacent domains can co-operate in ligand binding. Very recently, the first heterotypic ECM protein complexes have become available. Here, we review the advances of the last 5 years in understanding ECM protein structure, with special emphasis on those structures that have given insight into the biological functions of ECM proteins. PMID- 11852229 TI - Elastic energy storage in human articular cartilage: estimation of the elastic modulus for type II collagen and changes associated with osteoarthritis. AB - The viscoelastic mechanical properties of normal and osteoarthritic articular were analyzed based on data reported by Kempson [in: Adult Articular Cartilage (1973)] and Silver et al. (Connect. Tissue Res., 2001b). Results of the analysis of tensile elastic stress-strain curves suggest that the elastic modulus of cartilage from the superficial zone is approximately 7.0 GPa parallel and 2.21 GPa perpendicular to the cleavage line pattern. Collagen fibril lengths in the superficial zone were found to be approximately 1265 microm parallel and 668 microm perpendicular to the cleavage line direction. The values for the elastic modulus and fibril lengths decreased with increased extent of osteoarthritis. The elastic modulus of type II collagen parallel to the cleavage line pattern in the superficial zone approaches that of type I collagen in tendon, suggesting that elastic energy storage occurs in the superficial zone due to the tensile pre tension that exists in this region. Decreases in the elastic modulus associated with osteoarthritis reflect decreased ability of cartilage to store elastic energy, which leads to cartilage fibrillation and fissure formation. We hypothesize that under normal physiological conditions, collagen fibrils in cartilage function to store elastic energy associated with weight bearing and locomotion. Enzymatic cleavage of cartilage proteoglycans and collagen observed in osteoarthritis may lead to fibrillation and fissure formation as a result of impaired energy storage capability of cartilage. PMID- 11852230 TI - A structural model for force regulated integrin binding to fibronectin's RGD synergy site. AB - The synergy site on fibronectin's FN-III(9) module, located approximately 32 A away from the RGD-loop on FN-III(10), greatly enhances integrin alpha(5)beta(1) mediated cell binding. Since fibronectin is exposed to mechanical forces acting on the extracellular matrix in vivo, we used steered molecular dynamics to study how mechanical stretching of FN-III(9-10) affects the relative distance between these two synergistic sites. Our simulations predict the existence of an intermediate state prior to unfolding. In this state, the synergy-RGD distance is increased from 32 A to approximately 55 A, while the conformations of both sites remain unperturbed. This distance is too large for both sites to co-bind the same receptor, as indicated by experiments that confirm that increasing the length of the linker chain between FN-III(9) and FN-III(10) reduces alpha(5)beta(1) binding. Our simulations thus suggest that increased alpha(5)beta(1)-binding attributed to the synergy site, along with the associated downstream cell signaling events, can be turned off mechanically by stretching FN-III(9-10) into this intermediate state. The potential physiological implications are discussed. PMID- 11852231 TI - Real-time visualization of MMP-13 promoter activity in transgenic mice. AB - Cutaneous wound repair involves extracellular matrix degradation, cell migration, matrix resynthesis and tissue remodeling. In the rodent, transcriptional regulation of collagenase-3 (MMP-13) most likely plays a role in these processes. Therefore, we isolated and characterized a 1.76-kb 5'-flanking region of the mouse MMP-13 gene. Assay of promoter activity by transient transfection of HT1080 cells and primary mouse skin fibroblasts allowed identification of several functional regions of the 5'-flanking DNA. Expression of luciferase reporter constructs in these cells was induced by phorbol myristate acetate (PMA), but not by transforming growth factor-beta(2) (TGF-beta(2)). To study the regulation of MMP-13 in cutaneous wound healing, we generated transgenic mouse lines harboring the firefly luciferase reporter gene under control of a 660-bp mouse MMP-13 promoter which showed maximal response. MMP-13 mRNA levels in transgenic lung fibroblasts increased 1.5-2.6-fold after PMA challenge. MMP-13 promoter activity in wounds was visualized and quantified in vivo as luciferase bioluminescence. MMP-13 expression was present at day 1 and maximal at day 18 post-wounding. Luciferase activity progressed from the wound margin towards the center of the wound. In situ hybridization showed the same spatial and temporal patterns for the luciferase and endogenous MMP-13 mRNA. Both signals localized predominantly to dermal fibroblasts at the wound periphery but not to granulation tissue or to keratinocytes. These results suggested that MMP-13 participated in the wound healing of acute wounds, and it was a significant factor in long-term remodeling of wound connective tissue in rodent skin. PMID- 11852232 TI - Comparative analysis of the mouse and human genes (Matn2 and MATN2) for matrilin 2, a filament-forming protein widely distributed in extracellular matrices. AB - We previously identified matrilin-2 (MATN2), the largest member of the novel family of matrilins. These filament-forming adapter proteins expressed in a distinct, but partially overlapping, pattern in all tissues were implicated in the organization of the extracellular matrix. Matrilin-2 functions in a great variety of tissues. Here, we present the genomic organization of the highly conserved mouse and human MATN2 loci, which cover >100 kb and 167.167 kb genomic regions, respectively, and are composed of 19 exons. RT-PCR analysis revealed that alternative transcripts with identical protein coding regions are transcribed from two promoters in both species. The upstream, housekeeping type promoter is functional in all tissues and cell types tested. The activity of the downstream, TATA-like promoter preceded with putative motifs for the homeobox transcription factor PRRX2 is restricted to embryonic fibroblasts and certain cell lines. The oligomerization module is split by an U12-type AT-AC intron found in conserved position in all four matrilin genes. We assigned Matn2 to mouse chromosome 15, linked to Trhr and Sntb1 in a region synthenic to human chromosome 8q22-24. PMID- 11852233 TI - The effects of ascorbic acid on cartilage metabolism in guinea pig articular cartilage explants. AB - Ascorbic acid has been associated with the slowing of osteoarthritis progression in guinea pig and man. The goal of this study was to evaluate transcriptional and translational regulation of cartilage matrix components by ascorbic acid. Guinea pig articular cartilage explants were grown in the presence of L-ascorbic acid (L Asc), D-isoascorbic acid (D-Asc), sodium L-ascorbate (Na L-Asc), sodium D isoascorbate (Na D-Asc), or ascorbyl-2-phosphate (A2P) to isolate and analyze the acidic and nutrient effects of ascorbic acid. Transcription of type II collagen, prolyl 4-hydroxylase (alpha subunit), and aggrecan increased in response to the antiscorbutic forms of ascorbic acid (L-Asc, Na L-Asc, and A2P) and was stereospecific to the L-forms. Collagen and aggrecan synthesis also increased in response to the antiscorbutic forms but only in the absence of acidity. All ascorbic acid forms tended to increase oxidative damage over control. This was especially true for the non-nutrient D-forms and the high dose L-Asc. Finally, we investigated the ability of chondrocytes to express the newly described sodium dependent vitamin C transporters (SVCTs). We identified transcripts for SVCT2 but not SVCT1 in guinea pig cartilage explants. This represents the first characterization of SVCTs in chondrocytes. This study confirms that ascorbic acid stimulates collagen synthesis and in addition modestly stimulates aggrecan synthesis. These effects are exerted at both transcriptional and post transcriptional levels. The stereospecificity of these effects is consistent with chondrocyte expression of SVCT2, shown previously to transport L-Asc more efficiently than D-Asc. Therefore, this transporter may be the primary mechanism by which the L-forms of ascorbic acid enter the chondrocyte to control matrix gene activity. PMID- 11852234 TI - Matrix metalloproteinase activities and their relationship with collagen remodelling in tendon pathology. AB - Our aim was to correlate the activity of matrix metalloproteinases (MMPs) with denaturation and the turnover of collagen in normal and pathological human tendons. MMPs were extracted from ruptured supraspinatus tendons (n=10), macroscopically normal ("control") supraspinatus tendons (n=29) and normal short head of biceps brachii tendons (n=24). Enzyme activity was measured using fluorogenic substrates selective for MMP-1, MMP-3 and enzymes with gelatinolytic activity (MMP-2, MMP-9 and MMP-13). Collagen denaturation was determined by alpha chymotrypsin digestion. Protein turnover was determined by measuring the percentage of D-aspartic acid (% D-Asp). Zymography was conducted to identity specific gelatinases. MMP-1 activity was higher in ruptured supraspinatus compared to control supraspinatus and normal biceps brachii tendons (70.9, 26.4 and 11.5 fmol/mg tendon, respectively; P<0.001). Gelatinolytic and MMP-3 activities were lower in normal biceps brachii and ruptured supraspinatus compared to control supraspinatus (gelatinase: 0.18, 0.23 and 0.82 RFU/s/mg tendon respectively; P<0.001; MMP-3: 9.0, 8.6 and 55 fmol/mg tendon, respectively; P<0.001). Most gelatinase activity was shown to be MMP-2 by zymography. Denatured collagen was increased in ruptured supraspinatus compared to control supraspinatus (20.4% and 9.9%, respectively; P<0.001). The % D-Asp content increased linearly with age in normal biceps brachii but not in control supraspinatus and was significantly lower in ruptured supraspinatus compared to age-matched control tendons (0.33 and 1.09% D-Asp, respectively; P<0.01). We conclude that the short head of biceps brachii tendons show little protein turnover, whereas control supraspinatus tendons show relatively high turnover mediated by the activity of MMP-2, MMP-3 and MMP-1. This activity is thought to represent a repair or maintenance function that may be associated with an underlying degenerative process caused by a history of repeated injury and/or mechanical strain. After tendon rupture, there was increased activity of MMP-1, reduced activity of MMP-2 and MMP-3, increased turnover and further deterioration in the quality of the collagen network. Tendon degeneration is shown to be an active, cell-mediated process that may result from a failure to regulate specific MMP activities in response to repeated injury or mechanical strain. PMID- 11852235 TI - Analysis of self-assembly and apatite binding properties of amelogenin proteins lacking the hydrophilic C-terminal. AB - Amelogenins, the major protein component of the mineralizing enamel extracellular matrix, are critical for normal enamel formation as documented in the linkage studies of a group of inherited disorders, with defective enamel formation, called Amelogenesis imperfecta. Recent cases of Amelogenesis imperfecta include mutations that resulted in truncated amelogenin protein lacking the hydrophilic C terminal amino acids. Current advances in knowledge on amelogenin structure, nanospheres assembly and their effects on crystal growth have supported the hypothesis that amelogenin nanospheres provide the organized microstructure for the initiation and modulated growth of enamel apatite crystals. In order to evaluate the function of the conserved hydrophilic C-terminal telopeptide during enamel biomineralization, the present study was designed to analyze the self assembly and apatite binding behavior of amelogenin proteins and their isoforms lacking the hydrophilic C-terminal. We applied dynamic light scattering to investigate the size distribution of amelogenin nanospheres formed by a series of native and recombinant proteins. In addition, the apatite binding properties of these amelogenins were examined using commercially available hydroxyapatite crystals. Amelogenins lacking the carboxy-terminal (native P161 and recombinant rM166) formed larger nanospheres than those formed by their full-length precursors: native P173 and recombinant rM179. These data suggest that after removal of the hydrophilic carboxy-terminal segment further association of the nanospheres takes place through hydrophobic interactions. The affinity of amelogenins lacking the carboxy-terminal regions to apatite crystals was significantly lower than their parent amelogenins. These structure-functional analyses suggest that the hydrophilic carboxy-terminal plays critical functional roles in mineralization of enamel and that the lack of this segment causes abnormal mineralization. PMID- 11852236 TI - The PDZ domain of TIP-2/GIPC interacts with the C-terminus of the integrin alpha5 and alpha6 subunits. AB - Different cDNA libraries were screened by the yeast two-hybrid system using as a bait the cytoplasmic sequence of integrin alpha6A or alpha6B subunits. Surprisingly, the same PDZ domain-containing protein, TIP-2/GIPC, was isolated with either of the variants, although their sequences are different. Direct interaction assays with the cytoplasmic domain of the integrin alpha1--7 subunits revealed that in addition to alpha6A and alpha6B, TIP-2/GIPC reacted also with alpha5, but not other alpha integrin subunits. The specificity of the interaction was confirmed by in vitro protein binding assays with purified peptides corresponding to integrin cytoplasmic domains. Further analysis with either truncation fragments of TIP-2/GIPC or mutated integrin cytoplasmic domains indicated that the interaction occurs between the PDZ domain of TIP-2/GIPC and a consensus PDZ domain-binding sequence, SDA, present at the C-terminus of the integrin alpha5 and alpha6A subunits. The integrin alpha6B subunit terminates with a different sequence, SYS, which may represent a new PDZ domain-binding motif. PMID- 11852237 TI - CASH--a beta-helix domain widespread among carbohydrate-binding proteins. AB - In this article, we describe a novel, widespread domain (CASH) that is shared by many carbohydrate-binding proteins and sugar hydrolases. This domain occurs in more than 1000 proteins distributed among all three kingdoms of life. The CASH domain is characterized by internal repetitions of glycines and hydrophobic residues that correspond to the repetitive units of a predicted or observed right handed beta-helix structure of the pectate lyase superfamily. PMID- 11852238 TI - Designing a peptide carrier for efficient protein delivery. PMID- 11852243 TI - Evolution of enzyme cascades from embryonic development to blood coagulation. AB - Recent delineation of the serine protease cascade controlling dorsal-ventral patterning during Drosophila embryogenesis allows this cascade to be compared with those controlling clotting and complement in vertebrates and invertebrates. The identification of discrete markers of serine protease evolution has made it possible to reconstruct the probable chronology of enzyme evolution and to gain new insights into functional linkages among the cascades. Here, it is proposed that a single ancestral developmental/immunity cascade gave rise to the protostome and deuterostome developmental, clotting and complement cascades. Extensive similarities suggest that these cascades were built by adding enzymes from the bottom of the cascade up and from similar macromolecular building blocks. PMID- 11852244 TI - What does it mean to identify a protein in proteomics? AB - The annotation of the human genome indicates the surprisingly low number of approximately 40,000 genes. However, the estimated number of proteins encoded by these genes is two to three orders of magnitude higher. The ability to unambiguously identify the proteins is a prerequisite for their functional investigation. As proteins derived from the same gene can be largely identical, and might differ only in small but functionally relevant details, protein identification tools must not only identify a large number of proteins but also be able to differentiate between close relatives. This information can be generated by mass spectrometry, an approach that identifies proteins by partial analysis of their digestion-derived peptides. Information gleaned from databases fills in the missing sequence information. Because both sequence databases and experimental data are limited, a certain ambiguity often remains concerning which sequence variant(s) and modification(s) are present. As the common denominator of all the isoforms is a gene, in our opinion, it would be more accurate to state that a product of this particular gene rather than a certain protein has been identified by mass spectrometry. PMID- 11852245 TI - Reaction centres: the structure and evolution of biological solar power. AB - Reaction centres are complexes of pigment and protein that convert the electromagnetic energy of sunlight into chemical potential energy. They are found in plants, algae and a variety of bacterial species, and vary greatly in their composition and complexity. New structural information has highlighted features that are common to the different types of reaction centre and has provided insights into some of the key differences between reaction centres from different sources. New ideas have also emerged on how contemporary reaction centres might have evolved and on the possible origin of the first chlorophyll-protein complexes to harness the power of sunlight. PMID- 11852246 TI - Pho85 and signaling environmental conditions. AB - Through its association with a family of ten cyclins, the Pho85 cyclin-dependent kinase is involved in several signal transduction pathways in the yeast Saccharomyces cerevisiae. The responses mediated by Pho85 include cell-cycle progression and metabolism of nutrients such as phosphate and carbon sources. Although these responses require the phosphorylation of different substrates, and have different mechanistic consequences as a result of this phosphorylation, all appear to be involved in responses to changes in environmental conditions. Few of the activating signals or regulated targets have been unambiguously identified, but the kinase activity of Pho85 appears to inform the cell that the current environment is satisfactory. PMID- 11852247 TI - Reprieval from execution: the molecular basis of caspase inhibition. AB - The suppression of apoptosis is essential to the propagation of viruses, and to the control of development and homeostasis in insects and mammals. The central components of all apoptotic pathways are proteases of the caspase family. Therefore, it is not surprising that the processes of natural selection, as well as pharmaceutical chemists, have designed compounds that directly target caspase activity in attempts to regulate apoptosis. The mechanisms used by highly specialized naturally occurring caspase inhibitors (both host and viral) have remained obscure for some time. However, recently there has been significant progress in this field, particularly because of the structural elucidation of the complexes between caspases and an endogenous inhibitor (XIAP) and a viral inhibitor (p35). This article reviews the newly defined molecular basis for the regulation of the caspases by viral and endogenous inhibitors. PMID- 11852248 TI - Antifreeze proteins: an unusual receptor-ligand interaction. AB - Antifreeze proteins (AFPs) help organisms to survive below 0 degrees C by inhibiting ice growth. Although AFPs are structurally diverse, they typically present a large proportion of their surface area for binding to ice. Whereas earlier proposed binding mechanisms relied almost entirely on a hydrogen bond match between the AFP and ice, it now seems probable that van der Waals and hydrophobic interactions make a significant contribution to the enthalpy of adsorption. These interactions require intimate surface-surface complementarity between the receptor (AFP) and its ligand (ice). PMID- 11852249 TI - Protein Explorer: easy yet powerful macromolecular visualization. AB - Protein Explorer (PE, http://www.proteinexplorer.org) enables students, educators and other nonspecialists to visualize macromolecular structures easily. It also offers several advanced capabilities useful to protein structure specialists. Great attention has been given to making PE easy to use. Explanations, color keys and troubleshooting information are displayed automatically. There are also 'Frequently Asked Questions', a one-hour 'Quick-Tour', an alphabetical 'Help/Index/Glossary', and a detailed 'Tutorial'; all making PE much easier to use than either Chime or RasMol. Moreover, it is much more powerful; in addition to basic macromolecular visualization capabilities common to most similar programs, it offers one-click visualization of interfaces between moieties ('contacts'), cation-pi interactions and salt bridges, as well as easy-to-use routines to visualize regions of conservation in three-dimensional protein structures based on multiple sequence alignments. PMID- 11852250 TI - Dual site phospholamban phosphorylation and its physiological relevance in the heart. AB - Phospholamban (PLB) plays a primary role in regulating cardiac sarcoplasmic reticulum (SR) Ca(2+)-ATPase activity. Dephosphorylated PLB suppresses the SR Ca(2+) pump activity, whereas phosphorylation of PLB leads to deinhibition. A widely accepted sequential model of dual site PLB phosphorylation states that PKA dependent phosphorylation of Ser(16) is obligatory to phosphorylation of Thr(17) by Ca(2+)/calmodulin-dependent kinase II, and mainly accounts for beta-adrenergic receptor mediated cardiac relaxation. However, emerging evidence supports independent phosphorylation of Ser(16) and Thr(17) and their independent contributions to cardiac relaxation. Furthermore, concurrent activation of PKA and CaMKII signaling pathways exhibits a robust synergistic effect on phosphorylation of Thr(17), but not of Ser(16). Thus, the synergistic interaction may masquerade as a sequential phosphorylation of Ser(16) and Thr(17) under certain circumstances. Further studies are required to determine the exact process of dual site PLB phosphorylation and its functional roles in healthy and diseased hearts. PMID- 11852251 TI - Regulation of the endoplasmic reticulum calcium storage during the unfolded protein response--significance in tissue ischemia? AB - Endoplasmic reticulum (ER) is an organelle intimately involved in control of cell activities through Ca(2+) signaling, as well as in post-translational protein folding and maturation. Ca(2+) storage within the ER is required for both of these functions. Several of the ER-resident proteins essential for the protein folding pathway require Ca(2+) binding for their activity. A number of factors, including Ca(2+) depletion, may interfere with the folding pathway within the ER, with a potential for cell injury through an accumulation of malfolded protein aggregates. The Unfolded Protein Response involves a transcriptional upregulation of a number of the ER-resident folding helper proteins and becomes triggered when the folding pathway is blocked. To be effective, these upregulated proteins require a sufficient supply of Ca(2+) cofactor within the ER lumen. In tissue ischemia, where the availablity of this cofactor may be compromised, the newly described ability of the cell to boost the ER Ca(2+)-loading capacity by upregulating the ER Ca(2+) pump may be of particular importance for limiting cell injury and promoting survival. The novel focus on the pathophysiological significance of ER Ca(2+)depletion extends the scope of disturbed Ca(2+) homeostasis following ischemia beyond the consequences of the cytosolic calcium overload. PMID- 11852252 TI - Vascular endothelial growth factor and angiopoietins in focal cerebral ischemia. AB - A growing body of evidence indicates that vascular growth factors and their receptors are activated after stroke, and these factors may contribute to ischemic cell damage and to angiogenesis during recovery from stroke. In this review, we focus on recent evidence for roles of vascular endothelial growth factor (VEGF) and angiopoietin 1 (Ang 1) in blood-brain barrier (BBB) leakage and angiogenesis after focal cerebral ischemia. PMID- 11852253 TI - Etiological significance of apolipoprotein E mutations in lipoprotein glomerulopathy. AB - Lipoprotein glomerulopathy (LPG) is a newly recognized renal disease characterized by thrombus-like lipoproteins in the glomerular capillaries and abnormal lipoprotein profiles similar to those in type III hyperlipoproteinemia. Recently, these conditions have been shown to be associated with some apolipoprotein E (apoE) mutations. We found an apoE mutation (designated apoE Sendai) that substitutes arginine 145 with proline. This mutation occurs most frequently in Japanese patients with LPG. To elucidate the etiological role of this mutation in the apoE gene, we established an experimental model for LPG by transducing apoE-Sendai in apoE knockout mice with the use of an adenovirus vector. Based on the findings in patients with LPG and its animal model, we suggest that the glomerular lesions are not only caused by hyperlipidemia, but also by in situ interaction between lipoprotein-containing mutant apoE with the glomerulus. In this review, we outline the clinical features of LPG and discuss the relationship between apoE mutations and LPG. PMID- 11852254 TI - Emerging views of heparan sulfate glycosaminoglycan structure/activity relationships modulating dynamic biological functions. AB - Heparan sulfate glycosaminoglycans (HSGAGs) are an important subset of complex polysaccharides that represent the third major class of biopolymer, along with polynucleic acids and polypeptides. However, the importance of HSGAGs in biological processes is underappreciated because of a lack of effective molecular tools to correlate specific structures with functions. Only recently have significant strides been made in understanding the steps of HSGAG biosynthesis that lead to the formation of unique structures of functional importance. Such advances now create possibilities for intervening in numerous clinical situations, creating much-needed novel therapies for a variety of pathophysiological conditions including atherosclerosis, thromboembolic disorders, and unstable angina. PMID- 11852255 TI - Vasoregulation at the molecular level: a role for the beta1 subunit of the calcium-activated potassium (BK) channel. AB - Essential hypertension is among the most common and most costly medical conditions in the United States. Multiple defects in the kidneys, the vasculature, and the neuro-endocrine system may contribute to the development of this disorder. Within the past decade investigators have identified several molecular components of the vasculature that control tone and influence blood pressure. For example, the large conductance BK type calcium-activated potassium channel has recently been shown to play an important role in maintaining the dynamic equilibrium between vasoconstriction and vasodilation of vascular smooth muscle. Activation of vascular smooth muscle BK channels leads to hyperpolarization of the cell membrane, which causes deactivation of voltage dependent calcium channels and vasodilation. In this review, we will summarize recently published data focusing on the role of the BK channel's accessory beta1 subunit as well as other modulators of BK channel activation that influence vascular tone and blood pressure. PMID- 11852256 TI - The alpha(v)beta3 integrin, NF-kappaB, osteoprotegerin endothelial cell survival pathway. Potential role in angiogenesis. AB - The growth and survival of many cell types requires integrin-mediated adhesion to the extracellular matrix (ECM). Physiologically, the prerequisite of cell-ECM adhesion interaction for cell cycle progression and cell survival is likely to be important in tissue morphology and regression as a mechanism to regulate tissue architecture and cell number. Pathologically, anchorage-dependent survival may limit tumor invasion and metastasis. Endothelial cells are anchorage-dependent cells, and many ECM molecules interacting with different classes of integrins promote their survival. It has became clear, however, that during the angiogenesis process the alpha(v)beta(3) integrin plays a fundamental role in maintaining endothelial cell viability. The downstream signals regulating this process are becoming clarified, and new functions are described for molecules involved in apparently distant systems. PMID- 11852259 TI - Increasing the sophistication of access measurement in a rural healthcare study. AB - This paper considers the problem of deriving realistic access measures between population demand and health service locations, in the context of a rural region of England. The paper reviews approaches used in earlier work by the authors and others, and considers new public transport information systems that are now becoming available. An application is presented which incorporates the modelling of both private and public transportation travel times for access to district general hospitals in Cornwall. This information has been assembled from published timetables in order to evaluate the use of more sophisticated access measures that might be used when such data becomes more generally available. The work is set within the context of an ongoing substantive research programme concerned with health outcomes in the rural South West of England. PMID- 11852257 TI - Vascular progenitors: from biology to treatment. AB - The identification of endothelial progenitor cells (EPCs) in the adult human has forced us to reconsider how new blood vessels grow in physiological and pathological conditions in the adult human. An important question in angiogenesis research is to what extent these progenitors functionally contribute to revascularization of ischemic tissues and and to what extent they can be used for therapeutic angiogenic cell transplantation. Even more challenging is the concept that hematopoietic and other bone-marrow-derived stem cells might be recruited in the context of ischemia to induce neovessel formation. This review discusses some of the recent insights and outstanding questions on EPCs, both from a biological and therapeutic perspective. PMID- 11852260 TI - Population production and modelling mortality--an application of geographic information systems in health inequalities research. AB - Much research into spatial inequalities in health has sought the balance between compositional and contextual influences on observed patterns. Research published recently by the authors sought to determine whether composition of areas alone might account for the changing geography of mortality in Britain, between 1983 and 1993. The research required data describing Britain in terms of the numbers and location of people with every possible combination of age group, sex, social class and employment status. This paper describes the approach and the custom written geographic information system which estimated these data, and their subsequent application. PMID- 11852261 TI - Life and death of the people of London: a historical GIS of Charles Booth's inquiry. AB - Social reformer Charles Booth undertook a massive survey into the social and economic conditions of the people of London at the end of the 19th century. An important innovation of his Inquiry was the construction of large, detailed maps displaying social class of inner London on a street-by-street basis. These provide a detailed and vivid picture of the geography of poverty and affluence at this time. These maps have been digitised, georeferenced and linked to contemporary ward boundaries allowing Booth's measurement of social class to be matched to the measurement of social class in the 1991 census of population and standardised mortality ratios derived for all causes of death in the survey area between 1991 and 1995. The social class data were used to derive an index of relative poverty for both time periods and a comparison of the geographies of relative poverty and their relationship with contemporary mortality was made. Although the overall standard of living had increased, the geography of poverty at the end of the 19th century was very similar to that at the end of the 20th century. Moreover, the geography of all causes of death for people over the age of 65 was more strongly related to the geography of poverty in the late 19th century than contemporary patterns of poverty. This relationship was also true for mortality for specific diseases that are related to deprivation in early life. The paper concludes that the spatial patterns of poverty in inner London are extremely robust and a century of change has failed to disrupt it. PMID- 11852262 TI - Using Internet GIS technology for sharing health and health related data for the West Midlands Region. AB - Recent government legislation highlights the need for co-operative working by government agencies to improve the overall health of people and to help reduce the existing health inequalities in England. To effectively tackle health inequalities, access to a range of timely and relevant data sets about a region is vital. The Multi-Agency Internet Geographic Information Service (MAIGIS) project is a 3-yr pilot project funded by the Public Health Development Fund to establish an interactive map-based web site for sharing health and health related data for the West Midlands Region (http://maigis.wmpho.org.uk). Data sets within the MAIGIS project follow three broad themes of health, socio-economic and environmental information. Data are made available by different organisations and shared using geography as the linking theme. This paper discusses the use of Internet GIS technology for sharing health and health related data based on the issues that arose during the formative period of the MAIGIS project. Issues such as data confidentiality, amalgamation and copyright are discussed and the technical development of the project is outlined. The links that MAIGIS has formed with other regional and national initiatives for the sharing of health and health related information are also presented. Finally, the future work programme for the MAIGIS project is summarised. PMID- 11852263 TI - Geographies of need and the new NHS: methodological issues in the definition and measurement of the health needs of local populations. AB - For a variety of purposes it is increasingly necessary to establish the health needs of local populations. Following a critique of existing proxies of need, this paper presents and evaluates an alternative which draws upon epidemiological evidence concerning the age, sex and social class distribution of morbidity in order to estimate the prevalence of specific conditions in designated populations. By way of demonstrating the insights to be gained through the use of these indicative prevalence rates, the paper considers the treatment of ischaemic heart disease in a sample of 539 practices and presents evidence regarding the significance of deprivation and rurality in determining health service use relative to needs. PMID- 11852264 TI - c-Fos expression as endogenous marker of lumbosacral spinal neuron activity in response to vaginocervical-stimulation. AB - The focus of this paper is to describe a method for the simultaneous stimulation of the vagina and uterine cervix and recording of vaginal contractions in the female rat. The influence of the estrous cycle on vaginocervical sensitivity was also investigated. The use of a latex balloon, inflated with water via a syringe and connected to a pressure transducer allowed us to record vaginal contractions, the intensity of which is an index of vaginal sensitivity, and to stimulate those spinal neurons involved in the reflex arc; at the end of the stimulation the deflated balloon was used as a probe to perform a vaginal smear in order to determine the stage of the estrous cycle at the moment of the experiment. Activated neurons were identified by Fos-immunocytochemistry. Light microscope counting of Fos-immunoreactive neurons at different stages of the estrous cycle permitted us to quantify the response to vaginocervical stimulation and to demonstrate that vaginocervical sensitivity changes significantly throughout the estrous cycle. This finding confirms the important role of sex hormones in the modulation and control of the sensitivity of the vagina and cervix, a part of the female genital tract which is implicated in a variety of neuroendocrine, behavioral and neural changes. PMID- 11852266 TI - Perineuronal surface acetylcholinesterase activity of a fine neural network stained histochemically and observed with backscattered electron imaging and X ray mapping methods. AB - Fresh myenteric plexus-longitudinal muscle preparations were dissected by cleavage of the ileum of guinea pigs. The plasma membrane remained impermeable to lipophobic histochemical agents and the perineuronal surface (non-somatic) acetylcholinesterase (AChE) activity of the neural network was exclusively stained by a modified Karnovsky's histochemical medium. The cleaved face of the myenteric plexus was directly exposed to the exterior and could be clearly seen with scanning electron microscopy. A high contrast image of the neural network in three-dimensional space was observed by backscattered electron imaging (BSEI) of the histochemical precipitate which was produced on the perineuronal surface. The Cu and Fe of the histochemical precipitate (cupric ferrocyanide) were detected by energy dispersive X-ray analysis (EDXA). Elementary X-ray mapping of Cu and Fe on whole specimens provided images of the histochemical precipitate which were completely superposable to those of the BSEI, or even finer than those of the BSEI. The present results confirm the presence of Karnovsky's reaction on the perineuronal surface of the neural plexus, and suggest a useful method for studying this extraneuronal AChE in neural networks. PMID- 11852265 TI - A method to induce manganese accumulation in the brain of the cirrhotic rat and its evaluation. AB - Cirrhotic patients tend to accumulate manganese in their brain, especially in basal ganglia. Manganese is a well-known neurotoxic metal, however, its effect in a condition such as liver damage has not been explored deeply due to the lack of a suitable experimental model. A method to induce manganese accumulation in the brain of the cirrhotic rat is described. Cirrhosis was induced by obstruction of biliary duct and simultaneous treatment with manganese in the drinking water (0.5 or 1 mg/ml) during 4 weeks. Metal brain accumulation was low in sham-operated rats with both of the Mn concentrations used. In contrast, manganese treatment to bile obstructed rats resulted in fourfold and eightfold metal increments in the 0.5 and 1 mg/ml Mn2+ concentrations, respectively. This method is useful to induce brain manganese deposition and to study its consequences. PMID- 11852267 TI - The use of a remote-controlled minivalve, carried by freely moving animals on their head, to achieve instant pharmacological effects in intracerebral drug perfusion studies. AB - Intracerebral drug-perfusion studies in animals can be very efficiently performed with the 'reverse-dialysis' procedure. In this procedure, drugs are delivered into the brain via an intracerebrally implanted microdialysis probe. Traditionally, in reverse-dialysis studies the flow of control and drug solutions in the microdialysis site is alternated by large and heavy valves placed far from the experimental animal. In this arrangement, the drugs travel from the fluid alternating device for a long (20--60 min) period before reaching the brain. This can obscure the onset of drug action, makes it difficult to deliver drugs into the extracellular space during short-lasting behavioral episodes, and considerably limits the number of drug solutions that can be perfused within an experimental session. This report describes the use of a miniature (15 mm long and 8 mm diameter), lightweight (1.4 g) minivalve (patent pending) for combined neuronal recording--intracerebral microdialysis studies in freely moving rats. The device is activated remotely and carried by the animals on their head. This allows the experimenter to alternate the control and drug solutions in the intracerebral recording/dialysis site rapidly and to detect the drug-induced neuronal firing pattern changes instantly, without interfering with the animal's behavior. It is demonstrated that with this novel device the onset of drug actions on hippocampal neurons can be clearly defined and that these actions occur within 2 min after minivalve activation. Furthermore, it is demonstrated that the minivalve allows one to test a large number of drug solutions, successively, within the same experimental session. The described protocol offers a high-throughput method for testing the neuron-specific pharmacological effects of intracerebrally perfused drugs during various behaviors. PMID- 11852268 TI - The traffic light paradigm: a reaction time task to study laterally directed arm movements. AB - Patients with unilateral brain damage may show slowed or hypometric arm movements toward the contralesional space, as compared to movements directed towards the side of the brain lesion. The present article describes a reaction time paradigm devised to study accuracy and latency of directional arm movements in normal human subjects and brain-damaged patients. Experimental paradigms hitherto used to explore directional motor disorders often do not reliably disentangle between perceptual and motor factors, because they employ lateralized perceptual stimuli. The traffic light paradigm, instead, consists of visual stimuli presented on the vertical midline (like a traffic light) and hand responses to be produced in either hemispace. Thus, participants have to produce lateralized arm responses to central visual stimuli. Performance on this 'motor' paradigm can be contrasted with performance on a 'perceptual' reaction time task, consisting of similar, but lateralized visual stimuli and central motor responses. Results obtained with these paradigms on normal participants and brain-damaged patients are presented and discussed. These results give empirical support to the claim that the traffic light paradigm is suitable to study directional motor disorders in relative isolation from perceptual biases. PMID- 11852270 TI - A model of 'epileptic tolerance' for investigating neuroprotection, epileptic susceptibility and gene expression-related plastic changes. AB - Previous insult, for example, sustained epileptic seizures, confers a substantial temporary protection against the cellular damage induced by subsequent epileptic challenge. Here we describe a useful model of this so-called 'epileptic tolerance'. Expression of a status epilepticus was triggered by infusing the excitotoxic agent, kainate, into the right hippocampus of adult rats. An appropriate dose of kainate was used to cause brain damage in the homolateral, but not contralateral, hippocampus. At various times following this preconditioning insult, kainate was then re-administered into the lateral ventricle and neuroprotection was observed in the contralateral side between 1 and 15 days later. This model can be used to investigate the mechanisms of this endogenous neuroprotection. It is also particularly suitable for studying the epileptic susceptibility, as reflected by the modifications of the after discharge threshold, as well as any changes in gene expression induced associated with the preconditioning episode. PMID- 11852269 TI - Evaluation of neuronal cell death by laser scanning cytometry. AB - We developed a method in which laser scanning cytometry (LSC) is applied to evaluate cell viability. Neuronal cell death induced by glutamic acid, serum potassium deprivation and 3-nitropropionic acid was studied in cerebellar granule cells by neutral red assay (NR) and LSC, using propidium iodide (PI) as fluorescent dye. PI labeled the nuclei of dead neurons and increased fluorescence was measured using a laser scanning cytometer. Similar levels of damage for each injury were detected by NR or LSC. The protocol presented here, provides a fast and sensitive assay for the analysis of neuronal viability using LSC, and can be used to study new neuroprotective drugs in neuronal cell cultures. PMID- 11852271 TI - Neurogenic bladder model for spinal cord injury: spinal cord microdialysis and chronic urodynamics. AB - We describe an animal model to study neurotransmitter changes in parallel with urodynamic testing following Spinal Cord Injury (SCI). Urodynamic access was achieved using a subcutaneously placed 7 French dual lumen portacatheter. Spinal cord injury was induced by weight drop technique onto exposed dura at T8. The L6 S1 detrusor nuclei were localized stereotactically and microdialysis probe placement was confirmed through histologic methods. Chronic urodynamics revealed detrusor hyperreflexia (DH) 14 days following SCI. In vivo microdialysis of spinal cord amino acids was performed using CMA 11 (240 uM) probes in halothane anesthetized rats at baseline and intervals of 20-30 min following spinal cord injury. Significant increases in the excitatory amino acid glutamate, and the inhibitory amino acids, glycine and taurine, were seen following spinal cord injury. Amino acid levels peaked at approximately 40 min following contusion injury with glycine demonstrating the highest levels of all amino acids measured. This neurogenic rat model provides a useful means of examining the effects of spinal cord injury on bladder function. By utilizing spinal cord microdialysis, one could intervene at the level of the detrusor nuclei to modulate bladder function. PMID- 11852272 TI - Cochlear whole mount in situ hybridization: identification of longitudinal and radial gradients. AB - The morphology of the organ of Corti has a radial asymmetry and also changes longitudinally from base to apex. Cellular localization of transcripts within the inner ear has relied primarily on the use of sectioned tissue with in situ hybridization. However, radial and longitudinal gradients of expression are not readily recognized using sectioned tissue owing to problems in visualization of signals with varying intensities. Herein, we describe the use of whole mount in situ hybridization for identification of cochlear longitudinal and radial expression gradients in the neurosensory epithelium, hair cells. Not only can these hair cell gradients be shown in adult tissues, but also the developmental up-regulation and down-regulation of genes and their associated spatio-temporal expression patterns can be demonstrated. PMID- 11852273 TI - Application of real-time quantitative polymerase chain reaction to quantification of glutamate receptor gene expression in the vestibular brainstem and cerebellum. AB - Reverse transcription-polymerase chain reaction (RT-PCR) is a powerful tool to detect specific gene expression from a small amount of tissue, which is superior to the traditional RNA assays such as Northern blotting and in situ hybridization (ISH) in terms of sensitivity. However, conventional RT-PCR is not suitable for quantification due to its exponential nature. Recently, a real-time quantitative PCR method has been developed to overcome the weak points of RT-PCR, e.g. quantification. Here we describe the use of real-time quantitative PCR using a fluorescent TaqMan probe, to study the regional differences in expression of glutamate receptor subunit/subtype genes (NR1, NR2A, GluR2, KA2, mGluR1, mGluR7) in the central vestibular system including the vestibular nucleus complex, inferior olive and cerebellar flocculus. We found that real-time quantitative PCR yielded similar results to other techniques such as ISH but offered several advantages in terms of relative speed and ability to detect low levels of gene expression. We suggest that real-time quantitative PCR is a useful method to study gene expression for other neurotransmitter receptors in the vestibular brainstem and cerebellum, and is also expected to be more accurate to assess the changes in gene expression following any treatment. PMID- 11852274 TI - Working with a House of Commons Select Committee. PMID- 11852275 TI - Graduates without honour. PMID- 11852276 TI - Is incorporating CO2 'green'? PMID- 11852277 TI - The brighter sex? PMID- 11852278 TI - Pollen and inhalant allergy. AB - Anyone who suffers from hayfever will know what a serious problem allergies can be. In the long term, genetic technology may provide solutions in diagnosis, treatment and even avoidance. Eliminating allergens from pollen released by plants is technically feasible, but might this interfere with the natural processes of pollination? PMID- 11852279 TI - Marine biofouling: a sticky problem. AB - Organisms that colonise underwater surfaces, such as barnacle larvae and spores of algae, use a diverse array of biological 'glues' to provide both temporary and more permanent adhesion. The practical consequence of colonisation by these organisms is biofouling - something that has plagued mariners for years - causing increased drag and, in extreme cases, corrosion. Might there be a biological solution to this biological problem? PMID- 11852280 TI - Gone fishing: tool use in animals. AB - Chimpanzees impress and fascinate us with their use of tools, including twigs to 'fish' for termites and leaves to soak up liquids. But there are many intriguing examples of tool use described across the animal kingdom. Ants use grain to carry honey, and elephants can grip fly switches in their prehensile trunks. Even animals without limbs may use tools. PMID- 11852281 TI - The scourge of scabies. AB - Scabies ('Itch Mite') is truly a Great Neglected Disease that inflicts misery on millions. Molecular approaches, while still in their infancy, are providing a better understanding of the parasite and will have important implications for control and prevention. It has long been thought that dogs may act as a reservoir for human infections. However, genetic studies cast doubt over this supposition. PMID- 11852282 TI - The gene genie? AB - Gene therapy has the potential to cure currently incurable conditions, including some cancers and inherited disorders. It might even be used in the womb to prevent congenital abnormalities. The potential was greeted with great excitement ten years ago, when gene therapy first appeared to be viable, but little progress is perceived. Just how close are we to solving the obstacles in the way of successful gene implantation / replacement? PMID- 11852283 TI - The communications gap. AB - Gap junctions are ubiquitous channels that serve a vital function in communication between cells. Vertebrates and invertebrates have gap junction proteins that are very different at the sequence level but nonetheless have much in common? PMID- 11852284 TI - Chemical toxins and body defences. AB - Without realising it, we are all exposed to a wide range of toxins every day. Some are a natural part of our diet and environment and some are man-made. Over evolutionary time, biochemical pathways are developed that have allowed our bodies to cope with this onslaught. It is only in recent times, however, that we have really begun to understand how these mechanisms work. PMID- 11852285 TI - Future food. AB - Journalists and science fiction writers love to speculate that soon we will not be eating food as we have known it for millennia, but only the encapsulated products of the 'farmaceutical' industry. What nonsense! PMID- 11852286 TI - Cytokines and exocrine pancreatic cancer: is there a link? PMID- 11852287 TI - Cytokine gene polymorphisms in acute pancreatitis. AB - CONTEXT: Pro-inflammatory and regulatory cytokines play a key role in the pathogenesis of acute pancreatitis. Genetic loci encoding cytokines have been shown to be polymorphic, in some cases influencing protein expression. OBJECTIVE: To investigate if TNF and IL-10 gene loci are associated with the occurrence or severity of acute pancreatitis. SETTING: Acute surgical unit within large district hospital serving a population of 500,000. METHODS, Three TNF microsatellite loci (TNFa, TNFb, TNFc), the TNF-308 polymorphism, the IL-10.G microsatellite locus, and 3 i-allelic polymorphisms in the IL-10 5' region were typed using PCR in 135 acute pancreatitis patients and ethnically matched normal controls (n=107). Aetiology of disease was determined and patients grouped according to disease severity by assigning an organ failure score or classification according to the Atlanta system. MAIN OUTCOME MEASURES: Allelic frequency of polymorphic loci in patients with different aetiology and disease course in acute pancreatitis. RESULTS: No difference was noted in allelic frequency of any of the cytokine gene loci between groups stratified according to disease severity. When aetiology was studied again there was no significant difference in allelic frequency. CONCLUSIONS: The cytokine gene polymorphisms studied play no part in determination of disease severity or susceptibility to acute pancreatitis. PMID- 11852288 TI - Early prediction in acute pancreatitis: the contribution of amylase and lipase levels in peritoneal fluid. AB - CONTEXT: Predicting the severity of acute pancreatitis early in the course of the disease is still difficult. OBJECTIVE: The value of amylase and lipase levels in serum and peritoneal fluid might be of value in predicting the course of acute pancreatitis. DESIGN: Prospective study. PATIENTS: One-hundred and sixty-seven patients with acute pancreatitis as confirmed by computed tomography scan within 24 hours of admission were studied. MAIN OUTCOME MEASURES: Each patient was given an enzymatic score which reflected the predominance of serum or peritoneal levels of amylase and/or lipase. Enzymatic score was 0 if neither enzyme was predominant in the peritoneal fluid, 1 if amylase or lipase alone were predominant and 2 if both enzymes were predominant. The predictive value of the enzymatic score or computed tomography scan for a severe attack was determined. RESULTS: One-hundred and thirty-three attacks were graded as mild (79.6%) and 34 were considered as severe (20.4%). The frequency of severe acute pancreatitis significantly increased as the enzymatic score increased (5.4%, 12.5%, and 31.7% in 0, 1, and 2 enzymatic score patients, respectively; P<0.001). An enzymatic score greater than 0 predicted a severe outcome in 32 of 34 patients (sensitivity 94.1%, specificity 26.3%), whereas an enzymatic score of 2 predicted a severe attack in 26 of 34 patients (sensitivity 76.5%, specificity 57.9%). Edema on computed tomography scan was found in 97 of 129 mild attacks (specificity 75.2%) and necrosis in 25 of 33 severe attacks (sensitivity 75.8%), whereas all patients with severe attacks exhibited extrapancreatic acute fluid collection (sensitivity 100%, specificity 34.9%). CONCLUSIONS: Peritoneal dialysis is less predictive and more cumbersome than a computed tomography scan in the early prediction of acute pancreatitis. PMID- 11852289 TI - Pergolide in the treatment of patients with early and advanced Parkinson's disease. AB - Introduced on the market in 1989, pergolide, a D1/D2 dopamine receptor agonist, is still widely prescribed for the treatment of patients with early and advanced Parkinson's disease (PD). Initially, pergolide was introduced as an adjunct therapy to levodopa treatment in patients exhibiting fluctuating motor responses and dyskinesias. Results of recent randomized controlled clinical trials in de novo patients with PD show that pergolide is able to improve parkinsonian symptoms when used as monotherapy. Moreover, preliminary results of a long-term monotherapy study in early PD suggest that pergolide is as effective as levodopa, and that a significant delay in the time of the onset of levodopa-induced motor complications can be obtained. A number of randomized studies have shown that pergolide is more effective than bromocriptine as adjunct therapy to levodopa in patients with advanced PD; the greater benefit found with pergolide could be ascribed to its action on both D1 and D2 dopamine receptors. However, controlled comparative studies with new dopamine agonists, such as ropinirole, cabergoline, and pramipexole, have not been performed yet. Interestingly, few open studies in patients with complicated PD have shown that high doses of pergolide (> 6 mg/d) are able to improve motor fluctuations and dyskinesias through a dramatic reduction of levodopa dosage. The side-effect profile of pergolide is similar to that of other dopamine agonists, and complications such as sleep attack and serosal fibrosis have been rarely reported. PMID- 11852290 TI - Copolymer 1 (glatiramer acetate) in relapsing forms of multiple sclerosis: open multicenter study of alternate-day administration. AB - Daily 20-mg doses of Copolymer 1 have been shown to significantly decrease the relapse rate in patients with multiple sclerosis (MS). In the present open-label study, patients with relapsing MS were treated with the same dose of Copolymer 1 administered on alternate days. Sixty-eight patients were recruited: fifty-one and forty-one patients completed 1 and 2 years of treatment respectively. The relapse rate during the 2 years of treatment decreased by 80.8% compared with the 2 years before treatment (means, 0.56 +/- 1.02 versus 2.91 +/- 1.10, respectively; p < 0.0001). This lower rate is comparable with that obtained with daily open-label administration previously reported by the authors. The score on the Expanded Disability Status Scale did not differ from that at baseline after the first year of treatment, although it increased somewhat at the end of the second year (baseline = 2.72 +/- 1.55, 1 year = 2.71 +/- 1.59, 2 years = 2.97 +/- 1.80; p < 0.008). The drug was very well tolerated. This preliminary open-label study suggests that alternate-day therapy has beneficial effects and is well tolerated, comparing favorably with the effects of daily injections of Copolymer 1 in patients with relapsing MS. These results should be confirmed by randomized double-blind examinations. PMID- 11852291 TI - Therapeutic effects of an acetylcholinesterase inhibitor (donepezil) on memory in Wernicke-Korsakoff's disease. AB - Wernicke-Korsakoff's disease (WKD) is cognitively an amnestic state resulting from strategic lesions in the limbic system subserving the episodic memory network and resulting from thiamine deficiency. Neurochemical deficits have been implicated in the pathophysiology of amnesia based on the pathologic observations that various brainstem and basal forebrain nuclei are also affected. Previous treatment attempts with serotoninergic, noradrenergic, and cholinergic drugs have given controversial results. The objective of this study was to assess the effects of a cholinesterase inhibitor, donepezil, on memory, attention, and executive functions in patients with nonalcoholic WKD. Seven patients who developed WKD after a hunger strike were included in this single, blind, placebo controlled, one-way, crossover study. The patients were administered donepezil during the first 30 days, and were administered placebo during the following 30 days. Neuropsychological tests to evaluate verbal and visual memory, and attention and executive function were performed on days 0, 31, and 61. All patients completed both phases of the study. There were no statistically significant differences between the three evaluations, except for a difference between active treatment and the placebo phase during recall of the Rey Osterrieth complex figure, which was in favor of the placebo phase. There were no significant changes in favor of the active treatment. Cholinergic treatment with the cholinesterase inhibitor donepezil does not seem to provide marked beneficial effects in patients with WKD in this small, descriptive study. This may be because pathways mediating channel and state-dependent functions are impaired in this disease, and enhancement of state-dependent cholinergic transmission may not be sufficient. Subtle benefits, however, cannot be excluded because of the small sample size and the relatively short duration of the treatment. PMID- 11852292 TI - The SSRI, citalopram, improves bradykinesia in patients with Parkinson's disease treated with L-dopa. AB - Idiopathic Parkinson's disease (IPD) is characterized by motor signs such as akinesia, rigidity, and often tremor at rest. In addition to these symptoms, depression is a common finding affecting 40% of patients with IPD. This study evaluates the effect of the selective serotonin reuptake inhibitor, citalopram, on motor and nonmotor symptoms of depressed and nondepressed patients with IPD. Forty-six nondemented patients with IPD (24 men, 22 women; mean age 64 +/- 5.3 years; mean +/- SD disease duration, 6.4 +/- 3.2 years; mean +/- SD Hoehn-Yahr stage, 2.8 +/- 1.2) were included in the study. Patients were divided in two subgroups: depressed (n = 18) and nondepressed (n = 28). Citalopram was added in an unblinded manner, starting with 10 mg/d, and, after a week, increased up to 20 mg/d in the depressed subgroup (n = 18) and in half of the nondepressed subgroup (n = 14). Parkinsonian and depressive symptoms were evaluated before and after 1 and 4 months of treatment. Statistical evaluation was made by analysis of variance for repeated measures. Citalopram did not worsen motor performance in IPD, but improved bradykinesia and finger taps after 1 month and 4 months of treatment both in patients with and without depression (p < 0.05 versus baseline). A clear improvement in mood was also observed in 15 of 16 patients with depression. Although case reports indicate that citalopram can potentially worsen the motor symptoms in patients with PD, to date this effect has not been confirmed. Many of the symptoms, typically associated with depression, can be observed in nondepressed patients with IPD, because signs thought to represent depression can be produced by Parkinson's disease. In this study, we observed that when combined with levodopa, citalopram induces an improvement of motor performance, in particular of subscores 23 and 31 of Unified Parkinson's Disease Rating Scale both in depressed and in nondepressed patients with IPD. PMID- 11852293 TI - A double-blind crossover, placebo-controlled study of the adenosine A2A antagonist theophylline in Parkinson's disease. AB - Blockade of the adenosine A2A receptor potentiates the effects of levodopa in experimental animals and may offer a novel nondopaminergic target for drug therapy in Parkinson's disease (PD). Open-label trials suggest that the nonspecific adenosine antagonist theophylline improves parkinsonian symptoms and increases ON time in advanced patients with PD. In a double-blind, crossover, placebo-controlled trial, the authors investigated the ability of stable plasma levels of theophylline (between 10-20 microg/mL after 15 days of treatment) to modulate the long-duration response and the short-duration response of levodopa in 10 patients with PD. Although theophylline induced a longer duration of the effect of levodopa in all Unified Parkinson's Disease Rating Scale variables considered, including dyskinesias, maximal levodopa-induced improvement and the duration of the effect of levodopa did not differ significantly from placebo. Only the secondary variable "akinesia" showed a statistical tendency to a more prolonged beneficial response with theophylline during an acute levodopa test (short-duration response), and tremor worsened with theophylline during levodopa withdrawal (long-duration response). No differences were observed during the subacute course of study medication added to levodopa. During this exploratory study, the effects of theophylline were not strong enough to potentiate clearly the antiparkinsonian action of levodopa or to increase ON time in patients with advanced PD. PMID- 11852294 TI - Clinical evidence of an interaction between imipramine and acetylsalicylic acid on protein binding in depressed patients. AB - The binding of imipramine to plasma proteins was studied in 20 adult patients with endogenous depression, with the purpose of assessing the effect produced by its simultaneous administration with an analgesic. Patients were administered 150 mg/day imipramine for 5 days and the binding to plasma proteins was determined. This was repeated 2 days later, after simultaneous administration of imipramine with 1,000 mg/day acetylsalicylic acid (ASA). Adverse effects for each patient were registered during both phases and were classified as mild, moderate, or severe. Results showed 84.4 +/- 7.07% of imipramine bound to plasma proteins and 72.18 +/- 6.5% when imipramine was administered with ASA (p < 0.05). When imipramine was administered alone, 1.95 mild adverse events per patient were registered. When imipramine was administered with ASA, the mild adverse events increased to 3.1 (p < 0.01) and the severe adverse events increased from 0.6 to 1.5 (p < 0.01). The levels of free imipramine increased when ASA was administered, indicating a displacement on the binding to plasma proteins. When adverse events were compared for each treatment, the accumulation of the free fraction of imipramine caused an increase in adverse events as well as in their clinical severity. PMID- 11852295 TI - In vitro antioxidant properties of pentoxifylline, piracetam, and vinpocetine. AB - Oxygen-free radicals play an important role in several physiologic and pathophysiologic processes. In pathologic circumstances, they can modify and damage biologic systems. Because oxygen-free radicals are involved in a wide range of diseases (cerebrovascular, cardiovascular, etc.), scavenging these radicals should be considered as an important therapeutic approach. In our in vitro study, we investigated the antioxidant capacity of three drugs: pentoxiphylline (Sigma Aldrich, St. Louis, MO, USA) piracetam (Sigma Aldrich), and vinpocetine (Richter Gedeon RT, Budapest, Hungary). Phenazine methosulphate was applied to generate free radicals, increasing red blood cell rigidity. Filtration technique and potassium leaking were used to detect the cellular damage and the scavenging effect of the examined drugs. According to our results, at human therapeutic serum concentration, only vinpocetine (Richter Gedeon RT) had significant (p < 0.01) scavenging activity with a protective effect that increased further at higher concentrations. Pentoxiphylline (Sigma Aldrich) and piracetam (Sigma Aldrich) did not have significant antioxidant capacity at therapeutic concentrations, but increasing their concentrations (pentoxiphylline at 100-times, and piracetam at 10-times higher concentrations) led to a significant (p < 0.01) scavenger effect. Our findings suggest that this pronounced antioxidant effect of vinpocetine and even the milder scavenging capacity of pentoxiphylline and piracetam may be of value in the treatment of patients with cerebrovascular disorders, but merits further investigations. PMID- 11852296 TI - Activity-based diary for Parkinson's disease. AB - The objective of this study was to develop a Parkinson's disease diary that evaluates a patient's difficulties in performing activities as a substitute for the amount of "on"- and "off"-time and to assess its clinimetric qualities. In this study, 84 patients with Parkinson's disease kept a diary for 2 or 3 periods of 5 days. Daily, five items were recorded across 11 time periods. Patients simultaneously recorded "on-off" in the traditional way. The diary was easily understood, and median recording time was 5-10 minutes a day. Clinimetric analysis showed that the diary could be reduced successfully to 3 days, in which five items (walking, transfers, manual activities, dyskinesias, and sleep) with four response options (no, slight, moderate, and severe difficulty) were assessed seven times daily. Sumscores of the first three items accurately predicted being "on" or "off" in 93% of the cases, making separate scoring of "on" and "off" unnecessary. The diary was internally consistent and showed good reproducibility. Construct validity with external measures was adequate, and comparisons between patients grouped by disease severity and by degree of fluctuations revealed significant differences in the expected directions. Taken together, this Parkinson's disease diary has a sound clinimetric basis, provides information on the extent of perceived disability, and thereby accurately reflects the severity of "off"-periods and the variability of motor fluctuations. PMID- 11852297 TI - Epidural lipomatosis secondary to indinavir in an HIV-positive patient. AB - A human immunodeficiency virus-positive patient receiving indinavir therapy developed a slowly progressive paraparesis. Magnetic resonance imaging findings were consistent with epidural lipomatosis. On discontinuing indinavir, symptoms gradually remitted. Although indinavir, a protease inhibitor, is known to cause abnormal fat accumulation, to the best of our knowledge this is the first report of epidural lipomatosis. PMID- 11852298 TI - Side effects of long-term treatment with fluoxetine. AB - Depressive disorders are frequently managed with long-term use of antidepressant medication. Even though the newer generation of selective serotonin reuptake inhibitor antidepressants exhibits a more favorable short-term, side-effect profile, effects of chronic use of such drugs remain unknown. Considering the limited data available on long-term, selective serotonin reuptake inhibitor management, we report for the first time on two cases of late-onset adverse effects occurring 6 and 10 years after chronic-fluoxetine treatment in which patients experienced symptoms of restlessness, tension, agitation, and sleep disturbances. Symptoms resolved after reduction or cessation of the medication. Our case reports suggest the existence of a late-onset side-effect profile, which appears similar to acute side-effect symptomatology. Super sensitivity of the serotonin-related receptors may develop over the long-term and account for the phenomenon. Careful clinical monitoring is recommended to detect any late-onset, medication-related side effects. PMID- 11852299 TI - Pramipexole ameliorates neurologic and psychiatric symptoms in a Westphal variant of Huntington's disease. AB - Levodopa is recommended as a therapeutic approach for patients with the hypokinetic-rigid, Westphal variant of Huntington's disease, but no guidelines are available for the case of nonrespondence to levodopa. In this study, however, we report a 34-year-old woman with this rare variant who showed a clear-cut improvement in motor function and depressive symptoms after treatment with pramipexole, a new dopamine agonist. PMID- 11852300 TI - Breast tumor localization using sestamibi. AB - PURPOSE: The sensitivity of sestamibi scanning is 85% for breast lesions that measure >or=1 cm in diameter. This detection technique complements mammography and clinical examination and can benefit patients with very dense breast tissue. An innovation in nuclear imaging uses a lead marker for localization. MATERIALS AND METHODS: A 53-year-old woman underwent scintigraphy to clarify the indeterminate findings of a mammogram. Left breast biopsies 7 and 8 years earlier had yielded benign results. Mammography revealed a somewhat asymmetric stromal pattern, but the tissue appeared stable compared with results of previous studies. No focal abnormalities were identified. The original sestamibi breast scan revealed focally increased sestamibi uptake in the left breast. She was referred for another sestamibi scan because no radiographic or palpable abnormality correlated with the scintigraphic findings, and the lesion was believed to be nonlocalizable. Histologic examination revealed high-grade, poorly differentiated infiltrating ductal adenocarcinoma. RESULTS: After intravenous administration of Tc-99m sestamibi, the site of the lesion was identified using a lead marker, the persistence scope, and localization needles. This facilitated surgical removal. CONCLUSION: Using a lead marker allows placement of localization wires to guide surgical breast biopsy in patients whose lesions are visible by scintigraphy but not via mammography or palpation. PMID- 11852301 TI - The role of technetium-99m MAG3 renal imaging in the diagnosis of acute tubular necrosis of native kidneys. AB - The differential diagnosis for patients with acute renal failure of their native kidneys, as a result of primary intrarenal disease, includes acute tubular necrosis, glomerulonephritis, and interstitial nephritis. The role of MAG3 renography has not been studied in this setting. The authors describe four patients with acute renal failure in whom MAG3 renal imaging reliably identified acute tubular necrosis, as confirmed by follow-up kidney biopsies. In contrast to the poor parenchymal uptake observed in glomerulonephritis and interstitial nephritis, MAG3 shows a distinctive pattern in patients with acute tubular necrosis. For patients with acute renal failure, a renal scan can facilitate decision-making regarding the initiation of therapy. PMID- 11852302 TI - Ectopic ureteroceles in infants with prenatal hydronephrosis: use of renal cortical scintigraphy. AB - PURPOSE: This study assesses the contribution of renal cortical scintigraphy in the diagnostic evaluation of infants with ectopic ureteroceles. METHODS: The records of 20 infants (age range, 3 weeks to 4 months) who were referred for renal cortical scintigraphy after an ectopic ureterocele associated with a duplex kidney was found during follow-up of prenatal hydronephrosis were reviewed retrospectively. All infants underwent voiding cystourethrography, ultrasonography, and Tc-99m DMSA scintigraphy. RESULTS: All upper moieties showed absent or depressed Tc-99m DMSA uptake. Upper moiety functional impairment was associated with structural abnormalities shown by ultrasonography. The degree of functional impairment was reflective of ultrasonographically defined cortical thickness relative to pelvic diameter, but this relation varied. Eight of 20 (40%) lower moieties had diffusely depressed relative Tc-99m DMSA uptake. Focal defects were also present in three of these eight (38%) lower moieties. Lower moiety functional impairment was associated with higher degrees of pelvic dilatation and with vesicoureteral reflux. Ultrasonography revealed diffuse parenchymal thinning in four of the eight (50%) lower moieties with depressed uptake. Ultrasonography did not define focal parenchymal loss in any lower moiety. CONCLUSIONS: The degree to which function is impaired in the upper moieties of duplex kidneys with ureters terminating in ectopic ureteroceles is not highly predictable by ultrasonography. Lower moiety functional impairment and cortical defects are frequently present in these kidneys and often occur without ultrasonographic evidence of parenchymal damage. Use of renal cortical scintigraphy in the diagnostic evaluation of infants with ectopic ureteroceles is supported. PMID- 11852303 TI - Serendipitous detection of Cushing's disease by FDG positron emission tomography and a review of the literature. AB - A 70-year-old woman was referred for F-18 fluorodeoxyglucose (FDG) positron emission tomographic (PET) imaging of the brain to evaluate progressive dementia and neuropsychiatric symptoms. Although she had a history of hypertension and diabetes mellitus, she did not exhibit phenotypic features of Cushing's disease. The FDG-PET images revealed marked FDG uptake in the pituitary gland but no evidence of degenerative dementia. Two macroadenomas were confirmed by magnetic resonance imaging. Endocrinologic evaluation revealed Cushing's disease. After surgical resection of the tumors, the patient's symptoms decreased markedly. PMID- 11852304 TI - Postchemoembolization liver abscess: findings on hepatobiliary scintigraphy. AB - We describe a 41-year-old man with a 1-week history of nausea and vomiting 1 month after chemoembolization of a liver metastasis. The patient subsequently became febrile and developed right upper quadrant abdominal and midback pain. Findings of initial laboratory and imaging studies (a noncontrast computed tomographic [CT] scan and ultrasound) were not remarkable. Hepatobiliary scintigraphy, performed to rule out cholecystitis, revealed an abnormal area in the right lobe of the liver consistent with a focal bile leak into an abscess cavity. The patient was subsequently treated for liver abscess. In conclusion, hepatobiliary scintigraphy should be considered as a first-line test in the work up of patients whenever a postchemoembolization complication is considered likely. PMID- 11852305 TI - I-123 MIBG imaging and intraoperative localization of metastatic pheochromocytoma: a case report. AB - The authors describe the diagnostic use of I-123 MIBG scintigraphy in a 61-year old man who was thought to have a recurrence 25 years after a left adrenalectomy for a pheochromocytoma. Preoperative I-123 MIBG scintigraphy was performed twice along with intraoperative gamma probe localization of the lesions. The preoperative MIBG scintigraphy revealed three pathologic processes in the upper left abdomen, whereas computed tomographic scanning identified only one site of involvement. All three metastatic lesions were removed successfully with the aid of a gamma probe. Preoperative I-123 MIBG scintigraphy, combined with intraoperative gamma probe identification of I-123 MIBG foci, is feasible and a valuable tool to detect malignant masses possibly overlooked by other imaging techniques. PMID- 11852306 TI - An ectopic mediastinal parathyroid adenoma accurately located by a single-day imaging protocol of Tc-99m pertechnetate-MIBI subtraction scintigraphy and MIBI SPECT-computed tomographic image fusion. AB - PURPOSE: Because ectopic parathyroid adenoma (PA) is a frequent cause of failed initial surgery, an imaging approach with accurate preoperative localization is recommended by some authors in patients with primary hyperparathyroidism (HPT). METHODS: The authors describe a 52-year-old woman in whom primary HPT was diagnosed incidentally during a screening program for osteoporosis. The peculiarity of this case is that the patient was examined before operation in a single-day multimodal imaging protocol based on the combination of high resolution cervical ultrasound, planar Tc-99m pertechnetate-MIBI scans, and an MIBI-SPECT-computed tomographic (CT) image fusion study. An ectopic PA was accurately located in the upper middle mediastinum, close to the lower margin of the sternal notch. RESULTS: Guided by the MIBI-SPECT-CT fusion images, the surgeon performed a limited median sternotomy and easily removed the PA that was revealed before operation. To confirm the completeness of resection, a bilateral neck exploration was performed through the same incision, with identification of three normally sized parathyroid glands. CONCLUSIONS: Our experience suggests the utility of multimodality imaging procedures for the accurate preoperative localization of PAs, particularly when they are present in ectopic mediastinal locations. Such procedures, including the MIBI-SPECT-CT image fusion study, can be performed in a single day. PMID- 11852307 TI - Measurement of cerebral blood flow using graph plot analysis and I-123 iodoamphetamine. AB - PURPOSE: N-isopropyl-p[I-123]iodoamphetamin (IMP) is transiently taken up by the lungs after intravenous injection and its concentration in arterial blood varies depending on the degree of I-123 IMP uptake and subsequent washout. A method that does not require arterial blood sampling would be valuable to measure cerebral blood flow (CBF) using I-123 IMP. METHODS: The authors developed a new theory and a convenient new method of CBF determination using I-123 IMP that does not require blood sampling. Dynamic images of the head and chest were acquired immediately after intravenous injection of I-123 IMP in a series of 42 consecutive patients with cerebrovascular disorders or other brain diseases (31 men, 11 women; mean age, 58 +/- 11 years). Changes in the I-123 IMP counts of the regions of interest set in the head and pulmonary trunk were analyzed by the graph plot method, and the F values (CBF index slope) determined were compared with the mean CBF levels obtained by simultaneous autoradiography. RESULT: The F values correlated well with the mean CBF obtained by autoradiography (r = 0.818, P < 0.001). CONCLUSIONS: This innovative I-123 IMP graph plot analysis method using the time-activity curve of the head and pulmonary trunk alone is a noninvasive, convenient way to measure CBF. It is expected to become the most useful clinical technique for measuring CBF with I-123 IMP. This method can be used for patient follow-up and for comparing different patient groups evaluated in regional CBF studies. PMID- 11852308 TI - The possible role of F-18 FDG positron emission tomography in the differential diagnosis of focal pancreatic lesions. AB - PURPOSE: To compare the diagnostic values of different methods for the differentiation of malignant from benign pancreatic lesions. METHODS: In 22 patients with focal pancreatic lesions, the carbohydrate antigen (CA) 19-9 level was measured; abdominal ultrasound (US), computed tomography (CT), endoscopic retrograde cholangiopancreatography (ERCP), and F-18 fluorodeoxyglucose (FDG) positron emission tomography (PET) were performed; and the value of these methods were analyzed for their use in cancer diagnosis. RESULTS: Malignant lesions were identified in six patients and verified by surgery or clinical follow-up. The CA 19-9 level was elevated in four of the five patients examined (sensitivity, 80%). In all six cases, US and CT revealed hypoechogenic and hypodense areas (sensitivity, 100%). In one patient, ERCP was unsuccessful but yielded true positive results in three others (sensitivity, 60%). The sensitivity of FDG PET was 100%. Sixteen focal cases of pancreatic disease proved to be benign. The CA 19-9 level was elevated in four of them (specificity, 73%). Hypoechogenic and hypodense areas were evident on US and CT in eight patients. The specificity of CT was 50% (8 of 16 cases). The specificity of US was 47% (7 of 15 cases). The specificity of successful ERCP was 92%. Fourteen negative FDG-PET results were truly negative. In two patients, however, the PET findings proved to be falsely positive (specificity, 88%). CONCLUSIONS: FDG-PET is an effective tool to differentiate malignant from benign focal pancreatic lesions. In persons with focal pancreatic hypoechogenic or hypodense lesions detected by CT or US and an elevated CA 19-9 level, FDG PET should be the next step in the diagnostic strategy. PMID- 11852309 TI - Tc-99m depreotide scintigraphy of breast carcinoma. PMID- 11852310 TI - Tc-99m-labeled red blood cell scanning localizes anastomotic hemorrhage between the distal ileum and duodenal stump of an enteric-drained pancreas transplant. PMID- 11852311 TI - Bony metastases seen on scintigraphy with samarium-153. PMID- 11852312 TI - Hydrothorax in continuous ambulatory peritoneal dialysis: peritoneoscintigraphy in a case of spontaneous closure of pleuroperitoneal communication. PMID- 11852313 TI - F-18 FDG uptake in congestive splenomegaly. PMID- 11852314 TI - Positron emission tomography can reveal abdominal periaortitis. PMID- 11852315 TI - F-18 FDG positron emission tomographic image of an aortic aneurysmal thrombus. PMID- 11852316 TI - Diffuse soft tissue uptake of I-131 after post-thyroidectomy ablation therapy for thyroid cancer. PMID- 11852317 TI - Osteoid osteoma of the rib detected on bone scintigraphy: importance of pattern recognition. PMID- 11852318 TI - Tc-99m infecton and Tc-99m MDP imaging in osteomyelitis of the hip joint. PMID- 11852319 TI - Atrial septal defect with Eisenmenger's syndrome confirmed by Tc-99m MAA scintigraphy. PMID- 11852320 TI - Detection of a recurrent parathyroid carcinoma with FDG positron emission tomography. PMID- 11852321 TI - Postextrasystolic potentiation on quantitative gated SPECT. PMID- 11852322 TI - Tc-99m DMSA localization in infantile hemangioma. PMID- 11852324 TI - Current readings in nuclear medicine. PMID- 11852323 TI - Evaluation of persistent edema with lymphoscintigraphy after femoral artery injury. PMID- 11852325 TI - Principles of privileging and credentialing for endoscopy and colonoscopy. PMID- 11852327 TI - Management of patients with colorectal cancer and concomitant abdominal aortic aneurysm. AB - PURPOSE: The aim of this study was to determine the optimal management of patients with colorectal cancer and abdominal aortic aneurysm in the elective situation. METHODS: All patients with a history of colorectal cancer and abdominal aortic aneurysm between 1986 and July 2000 were identified, and charts of those with concomitant disease were reviewed. RESULTS: A total of 435 patients with available charts were reviewed. Eighty-three patients with concomitant abdominal aortic aneurysm and colorectal cancer were identified. In 64 patients the colorectal cancer was treated first, and 44 of these patients had an abdominal aortic aneurysm less than 5 cm in diameter (average = 3.8 cm). No abdominal aortic aneurysm ruptured in the postoperative period. Median delay to colorectal cancer surgery from diagnosis was four days. Twenty patients with abdominal aortic aneurysm of 5 cm or greater (average = 5.4 cm) were treated for colorectal cancer first. In two of these patients (with abdominal aortic aneurysms sized 5 and 6.4 cm), the abdominal aortic aneurysm ruptured in the early postoperative period. Median delay to colorectal cancer resection was eight days. Twelve patients had both abdominal aortic aneurysm and colorectal cancer treated at the same time. The average size of the abdominal aortic aneurysm was 6.4 cm. Median delay from colorectal cancer diagnosis to resection was 15 days. No documented cases of graft infection occurred in this group; median follow-up was 3.2 years. Seven patients underwent abdominal aortic aneurysm repair before resection of colorectal cancer; in two patients, colorectal cancer was found at the time of resection. The average size of abdominal aortic aneurysm was 6 cm and median delay to treatment of colorectal cancer was 122 days, a statistically significant longer delay than in the other two groups (P < 0.0001). CONCLUSION: In patients with colorectal cancer and abdominal aortic aneurysm of 5 cm or more, treatment of colorectal cancer first may result in life-threatening rupture, whereas treatment of abdominal aortic aneurysm first may significantly delay treatment of colorectal cancer. Concomitant treatment seems to be a safe alternative. If anatomically suitable, the abdominal aortic aneurysm may be considered for endovascular repair followed by a staged colon resection. The presence of an abdominal aortic aneurysm less than 5 cm does not affect colorectal cancer treatment. PMID- 11852328 TI - Normal female anal sphincter: difficulties in interpretation explained. AB - PURPOSE: Our aims were to quantify the nature, characteristics, and frequency of variations in female anal sphincter anatomy. METHODS: Nulliparous patients from the antenatal clinic and healthy volunteers of both genders were studied. Sphincter length was determined by the position of the puborectalis sling. Defects in the external anal sphincter were defined at each level and recorded in degrees. Cylindric longitudinal images of the endoanal scans were created by a three-dimensional-representation software package. Manometry was performed by a pull-through technique. RESULTS: Fifty-seven nulliparous patients and 18 healthy volunteers were included in the study. The mean age was 39 years for males and 28.35 years for females. There was no significant difference in overall sphincter length or in the internal anal sphincter length as a percentage of overall sphincter length between genders. All nine males had a complete ring of external anal sphincter along the full sphincter length. In the external anal sphincter below the level of the puborectalis sling, a natural gap occurred in 43 nulliparous (75 percent) and all 9 female volunteers. The greater the size of the defect, the greater its extent (mean 1.33 cm for >90 degrees and 1.16 cm for <90 degrees; chi-squared P = 0.008, eight degrees of freedom). Manometry provided confirmatory evidence of the gaps seen. Anal manometry was analyzed by Mann Whitney U test for continuous nonparametric data and t-test for comparison between genders. CONCLUSION: The female sphincter has a variable natural defect occurring along its anterior length. This makes interpretation of the isolated endoanal ultrasound difficult and explains previous overreporting of obstetric sphincter defects. PMID- 11852329 TI - Endosonographic anatomy of the normal anal canal compared with endocoil magnetic resonance imaging. AB - PURPOSE: This study was designed to clarify the sonographic anatomy of the normal anal canal by comparison with endoanal magnetic resonance imaging, to determine agreement between these imaging modalities and interobserver error in measuring layer thickness. METHODS: Three-dimensional endosonographic and endocoil magnetic resonance images of the anal canal were obtained in four males and five nulliparous females aged 22 to 34 years. Images were analyzed at similar levels throughout the canal using a graphics-overlay technique to compare sonographic with magnetic resonance images. Measurements were taken at one level for agreement analysis between modalities and for interobserver variability in the measurement of the thickness of the main anal canal layers. RESULTS: The muscularis submucosae ani, muscle bundles in the longitudinal muscle layer, and puboanalis were identified on sonography. The outer border of the external sphincter was demarcated by an interface reflection with ischioanal fat. Clarification of the external sphincter anatomy allowed excellent correlation (Ri = 0.96) for the assessment of thickness. There was excellent correlation for the interobserver measurement of the external and internal sphincters and the submucosal width on endosonography, but there was poor correlation for the longitudinal muscle (0.12). CONCLUSION: The overlay technique has improved endosonographic interpretation, and measurement of external sphincter thickness has been validated both by comparison with magnetic resonance and on interobserver agreement. PMID- 11852330 TI - Pudendal nerve terminal motor latency testing: assessing the educational learning curve: can we teach our own? AB - PURPOSE: Pudendal nerve terminal motor latency testing is useful as a diagnostic tool in fecal incontinence. It has also been used as a predictive factor in sphincteroplasty repairs. The technique is seldom taught and mastered in colorectal training programs. The purpose of this study was to assess a learning curve for teaching this procedure. METHODS: The student was a formally trained colorectal surgeon with no pudendal nerve terminal motor latency experience; the instructor has performed more than 3,000 pudendal nerve terminal motor latency studies. Fifty consecutive patients had manometry and pudendal nerve terminal motor latency testing. Both the student and instructor performed pudendal nerve terminal motor latency in a sequential fashion. Variables collected included pudendal nerve terminal motor latency, completion of test, time to complete test, and accuracy of the test. Variables were analyzed with paired t-test and chi squared analysis. RESULTS: The study group included 41 female and 9 male patients. The average age of the patients was 53 years. Bowel complaints included constipation in 16 and incontinence in 34 patients. Data were analyzed in their entirety and at ten-patient intervals. The student tended to record longer latencies (P < 0.001). This led to false-positive rates of 23 and 21 percent, respectively, for left and right pudendal nerve terminal motor latency. On average, the time to complete the procedure was three times longer for the student than for the instructor (P < 0.001). At ten-patient intervals, the pudendal nerve terminal motor latency difference between the two groups disappeared at the 41-to-50-patient mark. CONCLUSION: Given the proper learning environment, pudendal nerve terminal motor latency testing can be mastered in a relatively short period. On the basis of this study, our estimation is 40 patients are required for a student to master this technique. The novice examiner can expect false-positive results early in the learning curve, and these should be validated accordingly. PMID- 11852331 TI - Anal sphincter complex: reinterpreted morphology and its clinical relevance. AB - PURPOSE: Recent clinical studies on the anal sphincter complex have criticized the lack of reliable morphologic concepts. The purpose of this study was to determine the anatomy and histology of the anal sphincter complex with the help of undisturbed anatomic preparations. METHODS: The anal sphincter complex was studied in axial, sagittal, and coronal sections of human fetal, newborn, and adult pelves. RESULTS: The anal canal was surrounded by the internal sphincter, the longitudinal muscle layer, and an external sphincter that turned in to become continuous with the internal sphincter and with it to enclose the longitudinal muscle bundles. The classical tripartite subdivision of the external sphincter was not confirmed. The external sphincter seems not to be a complete circle in certain planes, neither in the male nor in the female. Sexual differences of the ventral part of the external sphincter were already present in fetuses. Large lamellated corpuscles were embedded within the interlacing smooth and striated muscles. Branches of the pudendal nerve innervated them. CONCLUSION: Our anatomic and histologic findings highly correlate with the results of magnetic resonance imaging and endosonography as well as with the physiologic findings. Furthermore, they are of great clinical importance for the understanding of sphincter defects during vaginal delivery and for anorectal operations in the adult as well as in the child. PMID- 11852332 TI - Early recovery after abdominal rectopexy with multimodal rehabilitation. AB - PURPOSE: Abdominal rectopexy without sigmoid resection is usually associated with a hospital stay of four to ten days. Recent developments have shown that a multimodal rehabilitation program with epidural analgesia and early oral feeding and mobilization will reduce hospital stay after colonic resection. The aim of this study was to describe the results after abdominal rectopexy with a multimodal rehabilitation program. METHODS: Thirty-one consecutive patients with rectal prolapse, median age 69 (range, 24-85) years and including 14 patients of American Society of Anesthesiologists physical status III to IV, were scheduled for abdominal rectopexy with a multimodal rehabilitation program including 48 hours thoracic epidural analgesia or patient-controlled anesthesia (3 patients), early oral nutrition and mobilization, and a planned two-day postoperative hospital stay. Follow-up was done at two months postoperatively. RESULTS: All patients except one tolerated normal diet and were mobilized to the same level as before surgery on the first postoperative day. Bowel movement was reestablished at a median of Day 2, and median postoperative hospital stay was three (mean, 4.4) days. At two months follow-up 16 percent were incontinent vs. 74 percent before surgery. Constipation was noted in 43 percent before surgery vs. 28 percent at two months follow-up. CONCLUSION: Median hospital stay after abdominal rectopexy may be reduced to three days with postoperative multimodal rehabilitation. PMID- 11852333 TI - Risk of lymph node metastasis in T1 carcinoma of the colon and rectum. AB - PURPOSE: Several recent reports of high local recurrence and lymph node metastasis in T1 carcinoma of the rectum prompted us to study the risk factors for lymph node metastasis in these lesions. METHODS: We reviewed the clinical records of 7,543 patients who underwent operative treatment for carcinoma of the colon and rectum from 1979 to 1995. Only patients with sessile T1 lesions who underwent colorectal resection were included in the study, yielding an analysis cohort of 353 patients. The following carcinoma-related variables were assessed: size, mucinous subtype, carcinomatous component, grade, site in colon and rectum, lymphovascular invasion, and depth of submucosal invasion. For the depth, the submucosa was divided into upper third (sm1), middle third (sm2), and lower third (sm3). Chi-squared tests and logistic regression were used to evaluate the variables as potential risk factors for lymph node metastasis. RESULTS: The incidence of T1 lesions was 8.6 percent. In the analysis cohort, the lymph node metastasis rate was 13 percent. Significant predictors of lymph node metastasis both univariately and multivariately were sm3 (P = 0.001), lymphovascular invasion (P = 0.005), and lesions in the lower third of the rectum (P = 0.007). Poorly differentiated carcinoma was significant univariately (P = 0.001) but not in the multivariate model. No other parameter was associated with a significant risk. CONCLUSIONS: T1 colorectal carcinomas with lymphovascular invasion, sm3 depth of invasion, and location in the lower third of the rectum have a high risk of lymph node metastasis. These lesions should have an oncologic resection. In a case of the lesion in the lower third of the rectum, local excision plus adjuvant chemoradiation may be an alternative. PMID- 11852334 TI - One-stage laparoscopic restorative proctocolectomy: an alternative to the conventional approach? AB - PURPOSE: There is significant concern in the current literature over the safety of laparoscopic techniques in removal of the entire colon and rectum. The purpose of this study was to examine the results of a one-stage laparoscopic-assisted restorative proctocolectomy in patients with mucosal ulcerative colitis and familial adenomatous polyposis in a single institution experience. METHODS: All patients who underwent laparoscopic-assisted one-stage restorative proctocolectomy (29 mucosal ulcerative colitis; 3 familial adenomatous polyposis) over a 24-month period were followed up prospectively for short-term and long term complications and functional outcome. RESULTS: There were 32 patients (17 males), with a median age of 32 years (range, 16-29 years). There were no conversions to open surgery. There were two intraoperative complications, an inconsequential rectal perforation during mobilization and one staple line misfire. There were 11 postoperative complications: 3 obstruction/ileus, 2 pouchitis, 2 wound infections, 2 strictures, 1 pelvic abscess, and 1 pouch leak (at the top of the "J"). Three patients required reoperation (1 temporary ileostomy, 1 lysis of adhesions, and 1 transpouch drainage). The median number of bowel movements was seven per day (range, 2-15). CONCLUSION: A one-stage laparoscopic-assisted restorative proctocolectomy can be performed effectively and safely. Given that techniques in laparoscopic large-bowel surgery are still evolving rapidly, the role of this operation in the surgical treatment of patients with mucosal ulcerative colitis and familial adenomatous polyposis is likely to expand in the near future. PMID- 11852335 TI - Superselective mesenteric embolization with microcoils in a porcine model. AB - PURPOSE: There is a lack of data regarding the degree of ischemic change that may occur in small and large bowel after superselective arterial embolotherapy with platinum microcoils. The purpose of this study was to gain a clearer understanding of the ischemic complications of superselective embolization of peripheral mesenteric arteries with microcoils by gross and histologic examination of postembolectomy bowel specimens at several time intervals in an adult porcine model. METHODS: Two-millimeter platinum microcoils with fibrils were superselectively deployed into six isolated mesenteric branch vessels in each of nine adult pigs. The animals were observed for two days (n = 3), one month (n = 3), or three months (n = 3) and then killed. At necropsy, intestines were examined for gross abnormalities. Sections of bowel containing microcoils were identified under fluoroscopy, resected, and evaluated histologically. RESULTS: A total of 54 microcoils were deployed into the distal arterial mesentery supplying the jejunum (n = 14), ileum (n = 26), and colon (n = 14) of nine adult pigs. Each animal received six microcoils. There were no clinical complications, and all pigs gained weight during their observation periods. Gross examination of the intestines did not reveal any evidence of acute or chronic ischemia. The coils were found in the distal arterial vasculature of the intestine. Histologic examination revealed mild superficial necrosis of villous tips in several samples; however, this finding is likely related to a fixation artifact rather than ischemic injury. There was no other histologic evidence suggestive of ischemic injury. CONCLUSIONS: Gross and histologic findings after superselective arterial embolotherapy demonstrated minimal changes in the bowel, and there were no significant clinical consequences to the animals. We conclude that transcatheter arterial embolotherapy in pigs is safe and may be applicable in the control of massive lower gastrointestinal hemorrhage in humans. PMID- 11852336 TI - Comparison of clinicopathologic characteristics and genetic alterations between microsatellite instability-positive and microsatellite instability-negative sporadic colorectal carcinomas in patients younger than 40 years old. AB - PURPOSE: Many studies have demonstrated that sporadic microsatellite instability positive colorectal cancers share several clinicopathologic features with hereditary nonpolyposis colorectal cancers, including right-sided location, young age of onset, characteristic histomorphologic features, and a good prognosis. The aim of this study was to define distinct clinicopathologic features of sporadic microsatellite instability-positive colorectal cancers and to compare genotypic characteristics between microsatellite instability-positive and microsatellite instability-negative colorectal cancers in a young group. METHODS: We analyzed 61 cases of young patients (<40 years old) with colorectal cancers for microsatellite instability at five mononucleotide and three dinucleotide repeats, loss of heterozygosity at APC and DCC, and K-ras and p53 mutations. Microsatellite instability status was correlated with molecular genetic factors and clinicopathologic parameters. RESULTS: Microsatellite instability positivity was detected in 19 (31.1 percent) of 61 cases. Allelic alterations in TGFbetaRII, BAX, and IGFIIR were observed exclusively in microsatellite instability-positive tumors (63.1, 26.3, and 26.3 percent, respectively). Microsatellite instability positive tumors exhibited a lower frequency of the p53 mutation (10.5 percent) than microsatellite instability-negative tumors (47.6 percent; P < 0.05). However, microsatellite instability status was not associated with APC or DCC allelic deletion or with the K-ras mutation. Microsatellite instability-positive colorectal cancers exhibited a proclivity toward proximal location, expansive growth pattern, and large tumor size (P < 0.05). Microsatellite instability positive colorectal cancers had lower preoperative serum carcinoembryonic antigen levels (P < 0.05), a less advanced stage at presentation (P < 0.05), and a tendency toward better prognosis (P = 0.051) than microsatellite instability negative colorectal cancers. However, there was no difference between microsatellite instability-positive and microsatellite instability-negative colorectal cancers regarding gross features, tumor grade, and extracellular mucin production. CONCLUSION: These results suggest that sporadic microsatellite instability-positive colorectal cancers in young patients have different histomorphologic features from microsatellite instability-negative colorectal cancers and hereditary nonpolyposis colorectal cancers, some overlap of genetic alterations on multistep carcinogenesis with microsatellite instability-negative colorectal cancers, and a tendency for better prognosis. PMID- 11852337 TI - Progressive duodenal adenomatosis in a familial adenomatous polyposis pedigree with APC mutation at codon 1556. AB - PURPOSE: In familial adenomatous polyposis, genotype-duodenal phenotype correlations have not been clearly understood. We identified the adenomatous polyposis coli gene mutation in a family pedigree with severe duodenal adenomatosis. METHODS: Among 53 familial adenomatous polyposis families, we found a pedigree composed of five affected members with severe duodenal adenomatosis. Clinical manifestations of the family members were reviewed. The adenomatous polyposis coli gene of four members were screened by polymerase chain reaction based single strand conformation polymorphism or protein truncation test. RESULTS: The family was characterized by sparse colorectal polyposis, osteomas, and epidermal cysts. However, there were intrafamilial variabilities in the occurrence of fundic gland polyposis, congenital hypertrophy of the retinal pigment epithelium, and desmoids. All the members had duodenal adenomatosis in their second or third decades, and the adenomatosis in three members progressed during surveillance. A frameshift mutation was found at codon 1556 of the adenomatous polyposis coli gene in two members, and the equivalent mutation was confirmed by protein truncation test in another two. CONCLUSIONS: Distal 3' mutation of the adenomatous polyposis coli gene seems to contribute to severe duodenal adenomatosis in familial adenomatous polyposis. Specification of the adenomatous polyposis coli gene mutation may be a clue for surveillance strategy for duodenal adenomatosis in patients with familial adenomatous polyposis. PMID- 11852338 TI - Effective management of posthemorrhoidectomy secondary hemorrhage using rectal irrigation. AB - PURPOSE: How to manage posthemorrhoidectomy secondary hemorrhage, a rare but serious complication, effectively remains controversial. This study evaluated the effectiveness of using rectal irrigation as an initial treatment for posthemorrhoidectomy secondary hemorrhage. METHODS: Among 4,880 patients on whom elective closed hemorrhoidectomy for symptomatic hemorrhoidal disease was performed, 45 (0.9 percent) developed posthemorrhoidectomy secondary hemorrhage. The 45 patients were divided into two groups based on the initial treatment in the stoma therapy room (n = 25) or in the operating room (n = 20). Patients in the stoma therapy room group were treated with rectal irrigation, whereas those in the operating room group were examined under anesthesia and the bleeding point (if any) was under-run using a suture. The two groups were then compared with respect to the cost-effectiveness of treatment, rehospitalization stay, and satisfaction with treatment. RESULTS: The two groups (stoma therapy room vs. operating room groups) were comparable with respect to the mean age of patients (44 vs. 38 years), interval of hemorrhage (9.4 vs. 7.8 days), and estimated amount of blood loss (560 vs. 520 ml). Bleeding effectively stopped in 22 (88 percent) patients in the stoma therapy room group but only in 12 (60 percent) patients in the operating room group (P = 0.010). The rehospitalization stay was three days in the stoma therapy room and 4.9 days in the operating room group (P = 0.016). In addition, the stoma therapy room group had a greater satisfaction rate than the operating room group did (80 vs. 10 percent, P < 0.001). Moreover, the average cost of treatment in the operating room group was six-fold higher than that in the stoma therapy room group. CONCLUSIONS: Our data suggest that rectal irrigation is an effective initial treatment for posthemorrhoidectomy secondary hemorrhage and offers a high rate of patient satisfaction with a reduced hospital cost. PMID- 11852339 TI - Dynamic transperineal ultrasound in the diagnosis of pelvic floor disorders: pilot study. AB - PURPOSE: Defecating proctography and more recently, magnetic resonance imaging have both been used for diagnosis in patients with pelvic floor dysfunction. This pilot study assessed the feasibility of dynamic transperineal ultrasound in a range of specific disorders affecting evacuation. METHODS: A protocol of sagittal and transverse transperineal imaging was established defining the infralevator viscera and soft tissues and the margins of the puborectalis muscle. Dynamic measurements were possible for the extent of puborectalis shortening, the anorectal angle, and the movement during straining of the anorectal junction. Calculations were made of the depth of demonstrated rectoceles, the posterior urethrovesical angle, and the movement of the urethrovesical junction. Diagnoses were confirmed by proctography (where appropriate) and clinical examination. RESULTS: Transverse images of the anal sphincter were comparable with those obtained using endoanal ultrasonography. Sagittal images permitted the measurement of puborectalis contraction and the anorectal angle comparable with those obtained during defecography. Cystoceles were able to be diagnosed during closure of the posterior urethrovesical angle and abnormal urethrovesical junction descent during straining. Rectoceles, peritoneoceles, enteroceles, and rectoanal intussusception were readily identified using dynamic transperineal ultrasonography. CONCLUSION: Dynamic transperineal ultrasound is a simple, noninvasive way to assess dynamically the interaction of the pelvic viscera and their relationship to the pelvic floor musculature in patients with evacuatory disorders and pelvic floor dysfunction. PMID- 11852340 TI - Assessment of ileoanal pouch inflammation by interleukin 1beta and interleukin 8 concentrations in the gut lumen. AB - PURPOSE: The cytokines interleukin 1beta and interleukin 8 have previously been shown to be present in mucosal biopsy specimens from inflamed ileoanal pouches. Our aim was to use the method of whole gut lavage fluid to measure cytokine concentrations and relate these to disease activity. METHODS: Forty-two patients with ulcerative colitis were recruited (23 males). Their ages ranged from 20 to 73 years (median 39). A questionnaire was completed and whole gut lavage, pouchoscopy, and biopsy were performed. RESULTS: Both interleukin 1beta and interleukin 8 were present in the whole gut lavage fluid of ileoanal pouches, with concentrations ranging from 4 to 143 pg/ml (median 6.3 pg/ml) for whole gut lavage fluid interleukin 1beta and from 18 to 1000 pg/ml (median 53.7 pg/ml) for whole gut lavage fluid interleukin 8. Whole gut lavage fluid interleukin 1beta and interleukin 8 were simultaneously detectable in 24 patients. These included the five patients with pouchitis, who had higher levels of interleukin 1beta (75 pg/ml vs. 8 pg/ml, P < 0.005) and interleukin 8 (668 pg/ml vs. 106 pg/ml, P < 0.005) compared with the rest of the patients with detectable cytokines (n = 19). The sensitivity of whole gut lavage fluid interleukin 8 (>200 pg/ml) in the diagnosis of pouchitis was 1, and the specificity was 0.86. There was a significant positive correlation of both whole gut lavage fluid interleukin 1beta and interleukin 8 with all the gut protein loss markers (immunoglobulin G, albumin, alpha1-antitrypsin). CONCLUSION: Cytokine interleukin 1beta and interleukin 8 concentrations, along with other parameters of inflammation, are raised in pouchitis in the whole gut lavage. The results also suggest a spectrum of severity of "pouchitis," with clinical pouchitis fulfilling Moskowitz criteria at the severe end of the spectrum. PMID- 11852341 TI - Development of a chronic colonic intubation model in rats for the study of luminal factors in colonic diseases. AB - INTRODUCTION: A rat model of long-term colonic intubation has been developed to facilitate the in vivo study of colonic biology. This study aims to characterize this model. METHODS: The effects of intubation and sham surgery on animal behavior and weight gain were measured and compared with unoperated controls. The reproducibility of the model was assessed by comparing complication and failure rates for three operators. The distribution and excretion of infused materials were studied using radiology and gas chromatography of feces, respectively. The effects of the colonic tube and infusions on the mucosa were assessed histologically. RESULTS: There was about 10 percent weight loss postoperatively, more marked in those rats undergoing more extensive surgery. Subsequent weight gain was similar in all groups, and no behavioral effects of surgery were noted. There were no differences in histologic appearances or proliferative indices among the groups. All three operators had similar complication and tube dislodgement rates. Radiologic examination showed even distribution of infusate regardless of fecal consistency. Infusion through the two tubes allowed the cecum and the distal bowel to be targeted differentially. The infusions did not alter the consistency of the fecal pellets or induce defecation. Gas chromatography at various time points after butyrate infusion showed a small, but statistically insignificant, rise in fecal excretion, representing less than 10 percent of the infused butyrate. CONCLUSIONS: A highly reproducible in vivo rat model, with an experimental life span of five to six weeks, has been achieved. Biological agents can be accurately delivered via the colonic tubes and are retained in the colonic segment of interest. This intubation model should provide a valuable tool in the future for in vivo studies of colonic biology. PMID- 11852342 TI - The management of cecal volvulus. AB - Cecal volvulus is second only to sigmoid volvulus in its frequency of occurrence. Diagnostic doubt is not uncommon in cecal volvulus; nonoperative decompression is rarely achievable; and if gangrene supervenes, mortality rises appreciably. Resection is mandatory for gangrene and a grossly distended, thin-walled cecum. Cecopexy and cecostomy seem less-effective and more morbid options than resection and anastomosis for viable bowel. However, their role needs reappraisal in the light of advances in minimally invasive techniques. PMID- 11852343 TI - Rectal perforation, retropneumoperitoneum, and pneumomediastinum after stapling procedure for prolapsed hemorrhoids: report of a case and subsequent considerations. AB - Stapling procedure is a new technique for the surgical management of third-degree hemorrhoids. Even if long-term experience has not been reported, this new technique is generating a lot of interest and its use is becoming more common in Europe. Some articles have just been published about severe adverse effects of this operation, and in the present article we describe a case of a life threatening complication that occurred with the use of the stapling technique for hemorrhoidectomy. A patient with perineal descensus and third-degree hemorrhoids underwent a stapling procedure for the treatment of hemorrhoids. Retropneumoperitoneum and pneumomediastinum developed on postoperative Day 2 and a colostomy was performed, allowing a quick recovery of the patient. After six months the colostomy was closed and bowel function restored. Our experience, taken together with some other cases previously published of severe complications after such an operation, suggests caution in the use of this new technique for the treatment of a benign disease. PMID- 11852344 TI - Ergotamine-induced anorectal strictures: report of five cases. AB - PURPOSE: Ergotamine tartrate suppositories are used for treatment of migraine headache attacks. Chronic abuse may lead to severe anorectal complications such as ulceration, stricture, and rectovaginal fistula. These complications are rare, and only sporadic reports may be found. Nevertheless, awareness of this entity on the part of prescribing physicians and treating colorectal surgeons is essential for a successful outcome, because withdrawal of this medication is an inherent part of treatment. PATIENTS: Five female patients were referred for treatment of symptomatic strictures of the anal canal and lower rectum. All of these patients admitted prolonged, nearly daily use of three to seven ergotamine tartrate suppositories. RESULTS: Three patients with severe stenosis of the anal verge and anal canal were treated by Y-V anoplasty, and two patients with circular stricture of the lower third of the rectum had balloon dilatations. In all patients the use of ergotamine suppositories was stopped, and alternative medication was instituted. Long-term follow-up (3-12 years) showed complete resolution of symptoms. CONCLUSION: In view of the availability of new effective drugs for treatment of migraine headache (serotonin agonists) and considering the potentially severe complications of chronic use of ergotamine, the use of this medication should be abandoned. PMID- 11852345 TI - Non-Hodgkin's lymphoma in a patient with probable hereditary nonpolyposis colon cancer: report of a case and review of the literature. AB - PURPOSE: Hereditary nonpolyposis colorectal cancer kindreds are frequently associated with cancers in various organs, including endometrium, stomach, and ovary. However, hematologic malignancy has rarely been reported in association with this cancer syndrome. We present here the case of a probable hereditary nonpolyposis colon cancer patient in whom non-Hodgkin's lymphoma developed after curative resection of colon cancer. Our experience with this rare case encouraged us to review the literature for reports indicating a possible relationship between these diseases. RESULTS: A 52-year-old male whose family history was consistent with the criteria for hereditary nonpolyposis colon cancer underwent right hemicolectomy for ascending colon cancer. Histologically the tumor consisted of adenocarcinoma that was moderately differentiated with mucinous foci and that invaded beyond the muscularis propria. Neither metastasis nor lymphoma was found in paracolonic lymph nodes. Eight months after surgery, the patient developed non-Hodgkin's lymphoma of T-cell origin involving the ileum and lungs. Both colon cancer and lymphoma frequently showed microsatellite DNA instability, sharing alteration in a locus of chromosome 7 (D7S501). CONCLUSION: A possible association of hematologic malignancy with hereditary nonpolyposis colon cancer reported in the literature, together with a report that MSH2-deficient mice are susceptible to malignant lymphoma, strongly supports the finding that this patient's lymphoma was related to hereditary nonpolyposis colon cancer. Overall, this case manifested a distinct clinical course similar to that observed in an animal model that is deficient in DNA mismatch repair machinery, thus providing scientific and clinical implications for understanding the molecular basis of these tumors and for critical management of hereditary nonpolyposis colorectal cancer patients, respectively. PMID- 11852346 TI - Sacral magnetic stimulation for pain relief from pudendal neuralgia and sciatica. AB - INTRODUCTION: Magnetic stimulation of the sacral nerve roots is used for neurologic examination. However, no one has reported therapeutic efficacy of pain relief from pudendal neuralgia with sacral magnetic stimulation. METHODS: Five patients with pudendal neuralgia or sciatica received 30 to 50 pulsed magnetic stimuli of the sacral nerve roots. The median age of the patients was 59 (range, 28-69) years; there were 3 females. RESULTS: Sacral magnetic stimulation immediately eliminated the pain. The pain relief lasted between 30 minutes and 56 days (median, 24 hours). Adverse effects were not observed. CONCLUSIONS: This pilot study indicates that magnetic stimulation of the sacral nerve roots may be a promising therapeutic modality for pain relief from pudendal neuralgia and sciatica. Further studies should be performed to determine the appropriate intensity and frequency, as well as the utility of a second course, of magnetic stimulation treatment. PMID- 11852347 TI - Use of endoscopic titanium stapler in rectal anastomotic stricture. AB - A quick and effective technique is described here for the treatment of rectal anastomotic stricture. It consisted of cutting the stricture using the Ethicon 35 mm Endoscopic Titanium Stapler. A patient was treated here as a day-surgery case. Good functional and anatomic results were achieved on follow-up. PMID- 11852348 TI - Anal pain? Explore the brain! PMID- 11852349 TI - [Thyroid and Chernobyl]. PMID- 11852350 TI - [Normal values of plasma methoxyamines in the setting of renal insufficiency and peri-operative stress. Consequences for the etiological diagnosis of hypertension]. AB - INTRODUCTION: HPLC plasma methoxyamines measurements are the updated technique for the diagnosis of adrenergic hypersecretion. Their reliability meets that of urinary measurements. Significance of increased values is not yet fully established for the etiological diagnosis of hypertension in some situations, especially in case of renal insufficiency and in the peri-operative period. The aim of this study is to define the "normal" range of the values of plasma methoxyamines in both of those conditions. PATIENTS AND METHODS: in a General and Endocrine Surgical Unit, 3 homogeneous group of 20 patients each have been studied: group 1, control (patients awaiting thyroidectomy); group 2, patients on maintenance hemodialysis submitted for hyperparathyroidism; group 3, patients submitted to digestive surgery. Measurements were done pre-operatively in group 1, pre and post-operatively in group 2, and post-operatively in group 3. RESULTS: in comparison to the control (11.8 nmol/l), we observed in group 2 a 18 fold increase preoperatively, and a 29 fold increase at post-operative day 1. In group 3, we observed a 2.3, 2.7 and 2 fold increase at post-operative days 1,2 and 3 respectively. All those results were statistically significant. CONCLUSION: Results of measurements of plama methoxyamines should always be matched to the serum creatinine levels. They are meaningful for the diagnosis of endocrine origin of hypertension only late after the early post-operative period. PMID- 11852351 TI - [Malt lymphoma of the thyroid and Graves'disease]. AB - Maltoma of the thyroid associated to Graves'disease is not such a frequently described association. We report a case of Maltoma of the thyroid gland in a patient with Grave's disease. The diagnosis was established by histological examination after total thyroidectomy. Slow enlargement of the thyroid gland despite a clinically successful antithyroid drug therapy could be considered as an advocating clinical manifestation. Associated anti-neoplastic chemotherapy and cervical irradiation resulted in good outcome with six years follow-up. A review is presented on treatment and prognosis of these tumors. PMID- 11852352 TI - [Propythiouracil and perchlorate effects on thyroid function in young and lactating mice]. AB - We have studied the effects of propythiouracil (PTU) or perchlorate (CIO(4)(-)), given to the mothers' drink from the 15(th) day of pregnancy until the day of sacrifice, on thyroid function of suckling mice. Antihyroid drugs (PTU or CIO(4)( )) provoked growth perturbations of young mice during studied ages from 6 to 18 days. A decrease of body weight was respectively as follows: 14 and 22% in 6 day old mice; 16 and 23% in 10 day-old mice; 18 and 18% in 14 day-old mice; 19 and 11% in 18 day-old mice. We have noticed an hypertrophy of thyroid glands of pups and their mothers caused by an increase of pituitary TSH. Thyroid follicles presented the aspect of hypothyroid animals with an increase of follicular number and vascularisation. Structural modifications confirmed biochemical results. In fact thyroid iodine contents decreased strongly in young as follow: 40 and 43% in 6 day-old mice; 51 and 50% in 10 day-old mice; 66 and 84% in 14 day-old mice; 54 and 89% at 18 day-old mice and in their mothers (50, 37%; 59, 54%; 75, 65% and 85, 72%) respectively after PTU or CIO(4)(-) treatment. A decrease of iodine thyroid gland was accompanied by an important fall of free thyroid hormones levels (FT3 and FT4) in young and adult mice. A decrease of thyroid hormonemia could explain the pups' growth perturbations. PMID- 11852353 TI - [New perspective in the medical management of primary hyperparathyroidism]. AB - Most of the medical proposed for patients with primary hyperparathyroidism are inefficient or dangerous, with the exception of oestrogens and physiological doses of vitamin D in selected cases. PTH analogs, calcimimetic drugs and anti PTH immunization now provide attractive management perspectives. PMID- 11852354 TI - [Foreign adopted children growth follow-up]. AB - For twenty years, Western Europe has taken part in a substantial development of international adoption. A consequent risk of precocious puberty affects little girls following such adoption. This diagnosis is still often unrecognized or passed off as a problem from the country of origin. It is important however not to delay treatment so as not compromise the final height. The start of puberty depends heavily on weight. Weight gain provokes the secretion of leptine, the latter increasing the frequence of GnRH peaks. Hence the radical change in diet which often experienced by the little girls adopted from developing countries and placed in rich countries, can provoke the onset of puberty. It is therefore necessary to follow very closely the height and weight curves of children adopted from overseas, particularly during the first year following adoption. The risk is as great as the need for an improved diet itself. Girls aged from three to nine at the time of their adoption comprise the group most susceptible to this disease. It is advised to have an X-ray taken upon their arrival to ascertain the age of the bones, allowing follow-up care in the case of a too rapid acceleration in growth. The treatment is the same for all central precocious puberty. PMID- 11852355 TI - [Fluorescence in situ hybridization (FISH): current developments and prospects in pathology]. PMID- 11852356 TI - [Infantile fibrosarcoma: a clinicopathological and molecular study of five cases]. AB - We report 5 cases of infantile fibrosarcoma (4 boys and 1 girl) whose average age was 5, 7 months (range 0 days to 14 months). The tumor was congenital in 4 cases. All tumors presented in the extremities (forearm, hand, thigh: 1 case, lower leg: 2 cases). Treatment was based on surgery (3 cases: amputation, 2 cases: local excision) with a favorable course in all cases, even those with marginal excision (follow-up ranging from 5 to 21 years). The lesions were characterized by dense monotonous cells growing in a fascicular pattern, with small necrotic areas and scattered lymphocytes. The mitotic index was high (average 8/10 high-power fields). ETV6-NK3 chimeric RNA was detected by reverse transcriptase polymerase chain reaction in two cases out four cases from paraffin-embedded tissue blocks. The infantile fibrosarcoma is a good prognosis tumor characterized by particular histological features and ETV6-NK3 gene fusion. PMID- 11852357 TI - [Detection of extra chromosomes 12 by fluorescent in situ hybridization (FISH) in ovarian stromal tumors. Study of 12 cases and review of the literature]. AB - Chromosomic aberrations play a major role in the initiation and the progression of benign as well as malignant tumors. In particular, trisomy 12 is frequently observed in female genitourinary tract tumors and constitutes a recurrent and often unique anomaly in stromal ovarian tumors such as fibrothecomas. Today, the genetic analysis of fresh or fixed solid tumors is enabled by the fluorescent in situ hybridization method (FISH). Using FISH and/or conventional cytogenetics, we analysed 12 ovarian stromal tumors (6 fibromas, 3 fibro thecomas and 3 thecomas). All of these tumors were benign and trisomy 12 was observed in all cases. Moreover, 3 cases presented trisomy and tetrasomy for chromosome 12 simultaneously. The high frequency of trisomy 12 in this tumor type suggests that this abnormality might be implicated in ovarian tumorigenesis. PMID- 11852358 TI - [Germ cell tumors of testis, current concepts]. AB - Histological pleomorphism of testicular germ cell tumors complicates diagnosis. These tumors share highly specific differentiation properties ranging from intra tubular neoplasia to classical infiltrating lesions. This general review presents the histopathological features and diagnostic criteria of typical forms and atypical variants. In current practice, distinction between seminoma and non seminomatous germ cell tumors is fundamental for treatment. Histological typing of non-seminomatous tumors may be problematical, especially in "mixed" forms. Vitelline tumor variants and atypical embryonic carcinoma are described. Unlike scattered isolated syncytiotrophoblastic cells, true chorio-carcinoma is rare and must be carefully identified. Such cells are frequently associated with seminoma, embryonic carcinoma and vitelline tumors. Intra-tubular germ-cell neoplasia and gonadoblastoma, are pre-invasive lesions associated with cryptorchidism and gonadal dysgenesis. Teratomatous elements usually occur in mixed germ cell tumors. Rare pure teratoma are benign in children. On the contrary, post-pubertal forms have an unpredictable evolution. PMID- 11852359 TI - [Diagnostic pitfalls in infectious disease pathology]. AB - Numerous images observed in histopathology and cytopathology can mimic pathogens such as mycotic agents, parasites (protozoa or helminthes), bacteria or virus. An error of diagnosis may provoke the administration of a non efficient treatment which can also be toxic. The present review describes these false pathogens which can correspond to exogenous or endogenous agents. Basic morphological analysis of mycotic agents, parasites, bacteria and cytopathogenic viral effects are successively presented. Then, the main diagnostic pitfalls which can be observed during these infections are exposed. Finally, diagnostic problems occurring in case of contamination are rapidly cited. PMID- 11852360 TI - [Tracheopathia osteochondroplastica associated with a liver hydatic cyst broken in bronchi]. AB - Tracheobronchopathia osteochondroplastica (TBOC) is an unusual respiratory disorder characterized by cartilaginous or bony mucosal nodules in the tracheobronchial tree. It mainly affects men over 50 years old and clinical manifestations are observed when obstructive or infectious complications occur. We report a case of tracheopathia osteoplastica of the upper third of the trachea in a young 35-year-old mal, fortuitously discovered while exploring a biliptysis caused by a broken hydatic cyst of the liver. TBOC can be associated with various metabolic, inflammatory and neoplastic disorders, but its pathogenesis remains unknown. A metaplastic origin is actually the main hypothesis proposed. PMID- 11852361 TI - [Malignant mesothelioma with osteoblastic heterologous elements]. AB - A 59 year-old man with a history of asbestos exposure presented with a right pleural effusion and a diffuse pleural thickening with focal calcifications on chest X-ray. Cytological examination of pleural fluid indicated malignant mesothelioma. A biopsy specimen showed malignant mesothelioma surrounding a fragment of mature bone. The patient was treated with intrapleural interferon, but relapsed 3 years later. A fresh biopsy specimen showed round tumor cells surrounding osteoid substance. Only ten cases of this rare variant of malignant mesothelioma with osteoblastic heterologous elements have been reported in the literature. The most difficult differential diagnosis is primary pleural osteosarcoma. PMID- 11852362 TI - [Mediastinal lymphatic filariasis]. AB - Wuchereria bancrofti gravid adult female worms were detected in a mediastinal lymph node in a 76-year old man. The patient also had lung carcinoma. A blood cell counts did not show any eosinophilia, and three consecutive night blood samples were also negative for microfilariae. Filariasis and neoplasm association was purely coincidental. Epidemiological and histopathological aspects of adult filariasis are described. PMID- 11852363 TI - [Mammary carcinoma arising in microglandular adenosis: a case report]. AB - Microglandular adenosis is an uncommon benign breast entity considered as a variant form of adenosis. The diagnosis is frequently made by the pathologist as it is often clinically asymptomatic. Carcinoma arising in microglandular adenosis is very uncommon. We report a case of carcinoma arising in microglandular adenosis in a 54-year-old woman. The immunohistochemical profile, especially S100 protein expression and absence of epithelial membrane antigen, was useful to recognize the microglandular adenosis and the carcinoma of alveolar architecture, while invasion was suggested by the basement membrane disruption highlighted with anti-collagen IV immunostaining. The good prognosis of carcinoma associated with microglandular adenosis points out the importance of distinguishing this lesion from other breast carcinomas. PMID- 11852364 TI - [Gastric epithelioid angiosarcoma, a biopsy diagnostic pitfall]. AB - We report the case of a 70-year-old man who presented a bulky gastric mass. The diagnosis of poorly differenciated adenocarcinoma was made on the biopsy. On surgical specimen, the final diagnosis of gastric angiosarcoma with secondary intestinal involvement was established. The patient did not respond to chemotherapy and died 8 months later. The diagnosis was difficult because of epithelioid areas which expressed both cytokeratin and vascular antigens. Human herpesvirus 8 previously detected in some soft tissue angiosarcoma was not found by polymerase chain reaction. PMID- 11852365 TI - [A peculiar tumor of the knee]. PMID- 11852366 TI - [A swelling of the wrist]. PMID- 11852367 TI - [A vaginal tumor]. PMID- 11852368 TI - [An unusual lymphoma]. PMID- 11852369 TI - [A voluminous intestinal mass]. PMID- 11852370 TI - Diabetes in Africans. Part 1: epidemiology and clinical specificities. AB - The prevalence of diabetes in African communities is increasing with ageing of the population and lifestyle changes associated with rapid urbanisation and westernisation. Traditional rural communities still have very low prevalence, at most 1-2%, except in some specific high-risk groups, whereas 1-13% or more adults in urban communities have diabetes. Type 2 diabetes is the predominant form (70 90%), the rest being represented by typical type 1 patients and patients with atypical presentations that require more pathophysiological insight. Due to the high urban growth rate, dietary changes, reduction in physical activity, and increasing obesity, it is estimated that the prevalence of diabetes is due to triple within the next 25 years. In addition, long-term complications occur early in the course of diabetes and concern a high proportion of patients, probably higher than in other ethnic groups, and that could be partly explained by uncontrolled hypertension, poor metabolic control and possible ethnic predisposition. The combination of the rising prevalence of diabetes and the high rate of long-term complications in Africans will lead to a drastic increase of the burden of diabetes on health systems of African countries. The design and implementation of appropriate strategy for early diagnosis and treatment, and population-based primary prevention of diabetes in these high-risk populations is therefore a public health priority. PMID- 11852371 TI - Glycaemic control and obesity are the major determinants of diabetic dyslipidaemia in Hong Kong Chinese. AB - OBJECTIVES: Dyslipidaemia plays a major role in the increased mortality in diabetes. Our aim was to address the quantitative abnormalities and determinants for lipid abnormalities in Chinese Type 2 diabetic patients. MATERIAL AND METHODS: In this study, we examined 1 279 Chinese type 2 diabetic patients and compared them with 959 non-diabetic control subjects. RESULTS: Of the 1 279 Type 2 diabetic patients, 588 (46.0%) were men and 691 (54.0%) were women. The mean age was 40.4 +/- 8.1 years (median: 41.0 years, range: 16-72 years). Compared to the 959 age- and sex-matched non-diabetic controls, diabetic patients were more obese, had higher blood pressure and adverse lipid profile characterized by high triglycerides and low high-density lipoprotein cholesterol. After adjusting for age, sex, smoking, obesity, use of lipid-lowering drugs and anti-diabetic agents, diabetic patients had higher risk of having hypertriglyceridaemia (>=2.3 mmol/L) and low high-density lipoprotein cholesterol (<0.9 mmol/L) than non-diabetic subjects. The corresponding odds ratios were 2.9 and 1.5, respectively. With multiple regression analysis (stepwise), dyslipidaemia was mainly associated with glycaemia, obesity and age in diabetic patients, and obesity, male gender and age in non-diabetic subjects. CONCLUSION: We have confirmed the high prevalence and increased risk of dyslipidaemia in Chinese type 2 diabetic patients. The long term significance of these lipid abnormalities and their synergism on clinical outcomes requires further evaluation. PMID- 11852372 TI - Prevalence of diabetes mellitus and impaired glucose tolerance in Benghazi Libya. AB - OBJECTIVES: To assess, by a house to house study, the prevalence of diabetes, impaired glucose tolerance (IGT) and their associated risk factors in Benghazi, Libya using 75 gram oral glucose tolerance test (OGTT) and ADA 1997 and WHO 1998 diagnostic criteria. MATERIAL AND METHODS: A multistage cluster sampling was used to select the study population. A total of 314 men and 554 women underwent a standard 75 grams OGTT. The response rate was 77.7% for males and 89.2% for females. RESULTS: The overall prevalence of IGT was 8.5% (95% confidence interval (CI) 5.8 - 11.3) (men 8.6% 95% CI 7.7-9.6, women 8.5% 95% CI 5.0 -11.9), and that of diabetes was 14.1% (95% CI 10.9-17.1) (men 16.3% 95% CI 14.5-18.3 women 13.0% 95% CI 10.0 - 16.1). Diabetes was present in 19.4% (95% CI 15.4-20.5) (men 22.7% 95% CI 20.2-25.4, women 17.6% 95% CI 14.1-19.1) in 30-64 years age range. Prevalence of diabetes was slightly higher in urban than in rural areas (14.5% vs 13.5%). The prevalence of newly diagnosed diabetes in urban and rural areas were 3.6% and 7.3% respectively and that of known diabetes were 10.9% and 6.3% respectively. Associated risk factors with diabetes and IGT were age, family history of diabetes, hypertension, BMI, WHR and serum cholesterol. CONCLUSION: Diabetes is emerging as an important public health problem in Libya and should rank very high in the priority list of health planners. PMID- 11852373 TI - Micro- and macrovascular complications and hyperhomocysteinaemia in type 1 diabetic patients. AB - OBJECTIVE: To determine the range of plasma homocysteine values in a cohort of type 1 diabetic subjects and to analyse the relationship between homocysteine levels and chronic degenerative complications. MATERIAL AND METHODS: 71 type 1 diabetic inpatients admitted for poor glycaemic control and/or treatment of complications, especially macroangiopathic, were included in this study. Chronic diabetic complications, smoking prevalence, as well as current use of drugs were recorded, alongside fasting plasma homocysteine. RESULTS: Age and diabetes duration were 51 (34-63) and 23 (13-32) years respectively (median [percentile 25 75]). HbA(1)c was 9.3% (8.2-10.5). Homocysteine was 9.2 (7.1-13.6) micromol/l, and 17% of patients had elevated homocysteinaemia on the basis of a laboratory cutoff value of 15 micromol/l. Folic acid and vitamin B(12) levels were within the normal range. Univariate statistical analysis showed a significant positive association between homocysteine and age (P<0.001), diabetes duration (P<0.001), systolic blood pressure (P<0.001), plasma creatinine (P<0.001), cholesterol/HDL-C (P=0.021) as well as with retinopathy (P=0.016) and all complications (P<0.001), and a negative correlation with folic acid (P=0.004) and creatinine clearance (P<0.001). Using a multiple regression analysis taking into account major variables, we confirmed an independent association of homocysteine with age (P=0.003), creatinine (P<0.001) and folic acid (P=0.014), but not with vascular complications. CONCLUSIONS: Increased homocysteine is present in 17% of a limited group of poorly controlled type 1 diabetic patients, and is associated with age, creatinine and folic acid levels. In this type 1 population, there was no independent correlation of homocysteine with vascular complications, in particular macroangiopathy. PMID- 11852374 TI - Trends in antihypertensive drug use among orally treated diabetic patients, in France between 1981 and 1992. Impact of guidelines and new drugs. AB - OBJECTIVE: To compare antihypertensive drug use between diabetics and non diabetics and to analyse the change between the two last decennial national surveys on health and medical care in France, in 1981 and 1992. MATERIAL AND METHODS: These surveys included respectively 12,725 and 13,887 adults over 25 years. Use of all antihypertensive drugs marketed in France was compared between orally treated diabetics and non-diabetics who were non-insulin and non-oral anti diabetic treated population. RESULTS: In 1981, while diuretics were the first treatment in both populations (53% in diabetics vs 60% in non-diabetics, non significant), central inhibitors (31 vs 16% respectively, p<0.001) were the second agent among diabetic patients instead of beta blockers in non-diabetic patients (13 vs 22% respectively, p<0.05). In 1992, beta blockers were also less used in diabetics (21 versus 33%, p=0.001). In contrast, ACE inhibitors were more used (43 versus 35%, p=0.06). In both surveys and populations, other classes (reserpin, vasodilator and alpha blockers) had minor place in the strategy. The frequency of combinations of antihypertensive drugs was similar in both surveys and populations. The preferred combination treatment was in both populations reserpin with diuretics in 1981 (7 vs 9%, non significant) and ACE inhibitors with diuretics in 1992 (18 vs 18%). CONCLUSION: Independently of guidelines, our data suggest that the strategy of hypertension treatment has changed last years and that prescriptions were more targeted for diabetic patients. PMID- 11852375 TI - Effects of clinical audit on the quality of care in patients with type 2 diabetes: results of the DIABEST pilot study. AB - OBJECTIVE: To improve the quality of diabetes care in general practice by the use of audit. MATERIAL AND METHODS: A prospective multicenter pilot study. Thirty five groups of ten general practitioners (GPs) have been set up throughout France on a voluntary basis. These groups were led by a steering committee that includes a diabetologist and a GP. Each group established a consensus on healthcare standards and carried out 2 data collections over a 12-month period, with adoption of corrective measures between these 2 collections. RESULTS: 309 (90.4%) of the 342 practitioners completed the first data collection, assessing the management of 3,125 patients over a year. Less than half the groups have reached the 80% reference threshold for quality of care with regard to foot examination, microalbuminuria, ECG, fundoscopy and assessment of diet. The audit also found no intervention within one year in case of poor glucose control for 47% of patients, unsatisfactory quality of HbA(1c) measurement and insufficient resources for diet and education. 226 (66.1%) practitioners completed the second data collection assessing the management of 2,248 patients. Comparison between the two phases of the audit showed significant improvements for all the indicators of the process of care (p<0.001). Quality of HbA(1c) measurement and diet assessment by GPs progressed (p<0.01). Outcomes of care also improved with respect to the proportion of patients with HbA(1c)<=8% (p=0.007), fasting glycaemia<1.40 g/l (p=0.05) and SBP<140 mmHg (p=0.02). CONCLUSIONS: This pilot study confirms the feasibility of using clinical audit at the national level. It seems to be an effective measure to improve the management of patients with type 2 diabetes in primary care. It is intended that the large-scale DIABEST study will address this issue. PMID- 11852376 TI - Antepartum and early postpartum predictors of type 2 diabetes development in women with gestational diabetes mellitus. AB - OBJECTIVES: This study aimed at identifying ante-partum and early post-partum (one year) clinical and metabolic characteristics capable of predicting the future development of type 2 diabetes in pregnant women of Mediterranea area affected by gestational diabetes mellitus (GDM). MATERIAL AND METHODS: Seventy GDM patients were evaluated: mean age during pregnancy, plasma glucose levels under OGTT (100 gr. glucose), fasting, 1-h post-prandial plasma glucose levels, HbA(1c) at the third trimester, gestational week of GDM diagnosis, insulin therapy, and weight gain were all taken into consideration. Some maternal risk factors such as pre-pregnancy BMI, and maternal and fetal outcome of index pregnancy were also assessed. One year after delivery in the same patients, BMI, fasting and 1-h post-prandial plasma glucose, plasma glucose and insulinemia under OGTT (75 gr. glucose) were measured. We focused our attention on women who presented type 2 diabetes 5 years after pregnancy or IGT and those who, one year after pregnancy, were normal. RESULTS: Five years after pregnancy 49 women were normal, 5 had developed type 1 diabetes and were not considered, 6 had developed IGT, and 10 type 2 diabetes. Analysis of variables during pregnancy showed that those variables predicting type 2 diabetes were pre-pregnancy BMI, gestational week of diagnosis, need for insulin therapy, obesity, and plasma glucose at 60' OGTT. Analysis of variables evaluated one year after pregnancy showed that BMI, fasting and post-prandial plasma glucose, plasma glucose at each point of the OGTT, and plasma insulin at 30' OGTT were predictive of the development of type 2 diabetes. Furthermore, age, post-partum fasting plasma glucose, and plasma glucose under OGTT post-partum were predictive of the development of IGT. Our data show for the first time that, also in a Caucasian Mediterranean population, markers of the future development of diabetes do exist, as reported in literature. They also stress the importance of correct identification of GDM patients, in order to screen those at greater risk of developing diabetes, for whom it is imperative to set up prevention programs. PMID- 11852377 TI - Effect on compliance, acceptability of blood glucose self-monitoring and HbA(1c) of a self-monitoring system developed according to patient's wishes. The ACCORD study. AB - OBJECTIVES: To test a blood glucose monitor developed upon diabetic's recommendations (Glucotrend Premium). Self-monitoring of blood glucose (SMBG) generates hope when introduced, however several studies questioned its efficacy and many diabetics judge it too constraining. MATERIAL AND METHODS: Thirty diabetes centres in France, for 6 months in 179 insulin-treated diabetics, using SMBG but non optimally and with HbA(1c) (>=130% of the upper limit). Randomisation to 3 groups: either their previous system (Group A), or to the Glucotrend Premium monitor with a memory to assess compliance (Group B), or to another monitor, new for the patient, and with a memory too, the One Touch Profile (Group C). At entry, and then at 3 and 6 months, patients had an acceptability and compliance questionnaire, HbA(1c), count of weekly hypoglycaemia, record of insulin doses and an assessment of the key compliance factors. RESULTS: HbA(1c) improved significantly in the 3 groups, more markedly in groups B (Glucotrend) and C (One Touch), e.g. - 0.6 +/- 1.1% (group A), - 0.9 +/- 1.2% (group B) and - 1.0 +/- 0.9% (group C) at M6. Acceptability was judged better for groups B and C, an additional benefit for Glucotrend: better accuracy vs laboratory blood glucose (C/L) determinations and a lower utilisation cost. Intermediate (lente) and regular insulin doses only significantly decreased (26% and 10% respectively) in group B (Glucotrend) despite a decrease in HbA(1c). Compliance (defined as 75-150% of recommended self-monitoring) improved within the 3 groups (from 34% to 65%), this improvement was maintained after month 3 (M6: 76%) only in group B (Glucotrend), vs a worsening in groups A and C (M6: 62 and 57% respectively). A better accuracy of C/L was observed with Glucotrend at M0, M3 and M6. CONCLUSION: SMBG has limits due to various causes, and to specific difficulties of this invasive and repetitive technique. The development of a system based on advices formulated by patients themselves, Glucotrend Premium, has resulted in a marked improvement on acceptability, compliance and glucose control. PMID- 11852378 TI - Management of diabetic foot lesions in hospital: costs and benefits. AB - OBJECTIVE: The aim of this study was to evaluate the direct and indirect costs associated with diabetic-related foot lesions in patients hospitalized in a department of Endocrinology and Diabetology (36 beds). MATERIAL AND METHODS: Direct costs included costs associated with hospital stay, laboratory testing, medical and surgical treatment. Lesions were classified in 3 groups (I, II, III) according to their importance (Wagner classification). RESULTS: Among 1,779 admissions for diabetes, 163 (i.e., 9.16%) were related to a foot lesion. The stay in hospital for these lesions reached 7,247 days over a 5-year period, with an average stay duration of 45 days per lesion. Mean length of hospital stay varied according to the importance of the lesion: 26.87 days, 48.25 days, 57.12 days for group I, II and III, respectively. The total cost amounted to 914,534.39 US dollars and the mean cost 5,610.64 US dollars. Mean cost was 3,326.76, 5,712.24, 7,399.74 US dollars for group I, II and III, respectively. Nearly 80% of the financial costs were due to hospital stay. Primary healing occurred in 59.09% of the patients (n=78); 17.4% (n=23) of the patients required major amputation, 14.4% (n=19) minor amputation. Death rate reached 9.1% (n=12). CONCLUSION: An increase in length of stay and costs with importance of the lesion was identified. The strategy of care of the diabetic lesions should be based upon the prevention of ulcer formation: it seems actually the best mean (the least expensive) in a developing country where financial resources are very limited. This prevention should be made through regular patient education, appropriately fitted shoes, and regular careful examination by a General Practitioner or a Diabetologist. PMID- 11852379 TI - Insulin therapy may increase blood pressure levels in type 2 diabetes mellitus. AB - OBJECTIVE: To assess the effects of insulin therapy on blood pressure levels in type 2 diabetes mellitus (T2 DM). MATERIAL AND METHODS: This is a retrospective analysis of clinical records of 313 T2DM patients (125 men and 188 women), excluding those with proteinuria or hypertensive diseases and those taking drugs that may influence blood pressure levels except antihypertensive therapy. Mean age was 56.3 +/- 11.7 years and mean duration of diabetes was 7.1 +/- 5.5 years. After one week of observation under diet and maximal doses of oral antidiabetic drugs, patients who did not improve their glucose control were changed towards insulin therapy (n=129) and formed the insulin treated group (ITG), those who improved their glucose levels were maintained under oral therapy (n=184) and formed the orally treated group (OTG). Blood pressure levels were compared between the two groups at baseline and after a mean follow-up period of 12.1 +/- 6.1 months. Hypertension was considered if patients were known and treated or if SBP >=140 mmHg and/or DBP >=90 mmHg. RESULTS: At baseline, patients in ITG were moderately older (58.4 +/- 11 vs 54.9 +/- 12.1 years, p<0.05), had a longer duration of diabetes (9.2 +/- 6.2 vs 5.7 +/- 5 years, p<0.01), a lower BMI (24.6 +/- 4.6 vs 28.8 +/- 6.6 kg/m(2), p<0.01) and a higher frequency of retinopathy (44% vs 31.1%, p<0.05). There was no significant difference regarding sex ratio, WHR, family history of hypertension, plasma levels of creatinine and lipid parameters. SBP, DBP and frequency of hypertension were similar in both groups at baseline. After follow up, insulin treated group exhibited higher levels of SBP (150 +/- 25.7 vs 138.6 +/- 27.1 mmHg, p<0.001) and DBP (84.1 +/- 13 vs 75.8 +/- 14.9 mmHg, p<0.001) than orally treated group. Progression rate of hypertension frequency was mildly but not significantly higher in ITG than in OTG (+21% vs +12%, p=0.08) and was associated with weight gain in ITG only. SBP increase was mildly correlated with weight gain (p=0.06). In ITG, higher values of BMI (> 27 kg/m(2)) at baseline were associated with the highest increases of blood pressure levels under insulin therapy. No significant relationship was found with insulin doses. CONCLUSION: Insulin therapy may contribute to the development of hypertension in T2DM obese patients. Additional prospective randomised studies are required for a better appreciation of such influence. PMID- 11852380 TI - Effects of short-term modest weight loss on fasting and post-prandial lipoprotein sub-fractions in type 2 diabetes mellitus patients. AB - OBJECTIVE: We assessed the efficacy of a modest weight loss (1.5 +/- 0.3 kg) and simultaneous rapid improvement in glycemic control on fasting an post-prandial lipoprotein sub-fractions in nine overweight (BMI=28 +/- 1.7 kg/m(2)) well controlled Type 2 diabetic patients (HbA(1c)=7.3 +/- 0.1%). MATERIAL AND METHODS: They followed a non-drastical hypocaloric balanced diet (1 561 +/- 39 kcal/day) over ten days in hospital. The fat content of the diet was significantly lowered from 96 +/- 12 g/day to 62 +/- 4 g/day (p<0.03). Plasma lipid and lipoprotein levels were measured in fasting and four hours after standard breakfast and four hours after standard lunch twice before and after ten days of hospitalization. The sub-fractions of very low density and low density lipoprotein were obtained by cumulative flotation ultracentrifugation. RESULTS: This weight loss reduced two well known independent cardiovascular risk factors such as the post-prandial glycemic excursions (p<0.05) and the post-prandial lipemia (p<0.05). Multiple linear regression analyses identified weight loss as an independent variable accounting for the ability to predict post-prandial capillary triglyceride clearance (p<0.05). Improvements in post-prandial glycemic excursions which was also entered as a parameter did not appear as a variable being able to predict these changes (p=0.4). In addition to the 23% improvement in post-prandial capillary triglyceride clearance (p<0.02), a decrement in post-prandial VLDL-2 triglyceride enrichment was found (p<0.05). Finally, fasting and post-prandial LDL-3 cholesterol levels were diminished (p<0.05) and the LDL-2/LDL-3 mass ratio post-prandial kinetics were improved (p<0.05). CONCLUSIONS: Even a modest weight loss in overweight, average controlled type 2 diabetic patients can achieve a significant improvement in two cardiovascular risk factors, namely post-prandial triglyceride excursions and the LDL-2/LDL-3 mass ratio kinetics independently from glycemic control improvements. PMID- 11852381 TI - [Direct cost of hospitalized care in diabetic patients in Lome (Togo)]. PMID- 11852382 TI - [Type 2 Diabetes and Short Form 36-items Health Survey]. AB - OBJECTIVES: This study explores the quality of life in patients with type 2 diabetes with the French Short Form 36-items Health Survey which involves eight health concepts: physical functioning, body pain, role limitations due to physical health problems, role limitations due to personal or emotional problems, general mental health, social functioning, energy and general health perceptions. MATERIAL AND METHODS: The French SF 36 was proposed to 282 diabetic patients, 70 years of age and under, randomly selected from the database of Social Security healthcare office in Lyon (total: 4 644 patients). 160 healthy controls, matched for age and sex, were enrolled. 12 questionaires were not analysed because of linguistic difficulties. RESULTS: The data show that quality of life is altered into T2D compared with the control population in all 8 dimensions of SF-36 items Health Survey. CONCLUSIONS: Quality of life is an interesting element to take into account for practitioners during management of diabetic patients, that could prove useful in order to obtain a better compliance of the patients. PMID- 11852383 TI - [Hyperglycemia: a predictor of poor prognosis in acute stroke]. AB - Hyperglycemia is common and involves up to 50% of the acute stroke patients. Several clinical studies have revealed that hyperglycemia is associated with a poor outcome in terms of mortality and neurological recovery. The results obtained from experimental studies have shown that hyperglycemia exacerbates the ischemic lesions and is associated with an increase of the edema and size of the infarct, as well as a decrease in the cerebral blood flow. In contradiction with animal models no increase in hemorrhagic transformation of the brain infarct after reperfusion is observed among patients with hyperglycemia, unless they are treated with rt-PA. Elevated plasma glucose levels (glycemia > 8 mmol/L) predict poor prognosis, irrespective of age, severity, or stroke sub-type. The benefit to treat a glycemia >=11 mmol/l has been established in terms of mortality for patients with myocardial infarction. Although the clinical benefit of plasma glucose lowering therapy remains unknown regarding acute stroke, based on current evidence, we recommend to treat moderate to mild hyperglycemia with glucose potassium insulin infusions. PMID- 11852384 TI - [ The mobilization of an entire discipline around a white book: why?]. PMID- 11852385 TI - [Hepatitis B vaccination and serious side-effects: sequence is not consequence]. PMID- 11852386 TI - [Has the time come to measure the hemodynamic response to pharmaco-logical treatments of portal hypertension?]. PMID- 11852387 TI - [Adenoma or focal nodular hyperplasia of the liver? A difficult pathological diagnosis]. PMID- 11852388 TI - [Hemodynamic tolerance and rapid hypertrophy of a hepatic graft corresponding to less than 30% of the ideal mass in pigs]. AB - BACKGROUND AND OBJECTIVE: Evaluation of a new pig liver transplantation technique for survival and hypertrophy of a small-sized graft by providing adapted and controlled venous portal flow. MATERIAL AND METHODS: [corrected] Twenty Large White pigs underwent heterotopic liver transplantation after a mesocaval shunt and ligation of the superior mesenteric vein downstream from the shunt. The donor to-recipient weight ratio was below 30%. Furthermore, recipient's biliary duct and portal vein into the hilum were tied. In a control group, no mesocaval shunt was performed and the graft received the entire splanchnic venous flow. RESULTS: The mesocaval shunt provided diversion of 60% of the splanchnic blood flow. The median survival of study pigs was 39 days (range: 8-98). Median serum bilirubin levels at 1 week were 12 micromol/L (range: 4-59). At autopsy, graft weight was increased to 2.7 times the initial weight and histological findings were normal. In the control group, all pigs died quickly from acute splanchnic congestion. CONCLUSION: In a model of heterotopic liver transplantation using small-sized grafts, complete diversion of mesenteric blood flow through a mesocaval shunt resulted in hemodynamic tolerance and hypertrophy of a graft corresponding to less than 30% of the ideal mass. PMID- 11852389 TI - [Coinfection HIV-HBV and the liver]. PMID- 11852390 TI - [Follow-up after resection for cure of colorectal cancer: searching for the missing rationale]. PMID- 11852391 TI - [Follow-up of patients with colorectal cancer resected for cure in the Herault area. A medico-economical study]. AB - Optimal modalities of surveillance of colorectal cancers (CRC) resected for cure have not been determined so far and the overall improvement of 5-year survival related to surveillance has not been demonstrated. AIM OF THE STUDY: To retrospectively evaluate modalities, results and costs of follow-up of patients during the 5 years following the resection for cure of CRC. METHODS: We studied medical and economical data from records of 256 patients registered in the cancer registry of the Herault area who underwent a potentially curative resection of CRC in 1992. We analyzed comparatively modalities of follow-up in patients who were followed according to recommendations from the 1998 French consensus conference (standard follow-up) and in those who had a simplified follow-up. We evaluated cumulative costs of follow-up. RESULTS: Nine patients died in the postoperative period. Recurrence rate was 27% (69 patients). Sixty-nine patients had a standard follow-up (30% of the 231 classified patients) and 162 patients (70%) had a simplified follow-up. The specific survival rate (taking into account only death related to CRC) 5 years after resection for cure was 75%. The 5-year specific survival rate after diagnosis of recurrence was 12% in the patients with recurrent disease within the 5 years after initial therapy. The 5-year survival rate after standard and simplified follow-up were 85% and 79%, respectively (P=0.25). Total cost of follow-up of the 256 patients was 1 085 507 French francs (FF). Mean follow-up cost per patient was 5 527 FF. Cost of the examinations not recommended by the consensus conference represented 30% of the expenses. Individual total cost of the follow-up of patients alive 5 years after the diagnosis of the recurrence was 120 356 FF. CONCLUSION: In Herault area, clinicians carried out in 70% of the patients a simplified follow-up and in 30% of the cases a reinforced follow-up in comparison with French recommendations. Survival rates were not significantly different between the 2 groups. PMID- 11852392 TI - [Minimally invasive surgery for diverticula of the thoracic esophagus. Results in 11 patients]. AB - OBJECTIVES: Surgical treatment of diverticula of the esophagus is associated with substantial mortality and morbidity. Few data have been published concerning results of minimally invasive surgery. The aim of the study was to retrospectively assess the results of minimally invasive surgery (either thoracoscopy or laparoscopy) in a first series of patients with diverticula of the thoracic esophagus. METHODS: Eleven consecutive patients with symptomatic thoracic diverticula of the esophagus were operated on between December 1992 and March 1999. Five were operated on by right thoracoscopy, 4 by laparoscopy and 2 by thoracoscopy and laparoscopy. The procedure performed varied according to the location and the macroscopic aspect of the diverticulum, as well as of the associated disorders (gastroesophageal reflux, hiatal hernia and/or motor disorders). RESULTS: Postoperative mortality was nil. Three patients developed an esophageal fistula; one with an esophago-bronchial fistula required another operation. Postoperative pain was treated with morphine (median duration 4 days) or IV paracetamol (5 days). Long term results were excellent in 1 patient, good in 6, fair in 2 and poor in 2. These 2 latter patients were operated on another time. One of them was operated on 3 years later for aperistalsis of the esophagus and the other one was operated 4.5 years later for paraesophageal hernia; late results of these operations were fair. CONCLUSION: These results suggest that minimally invasive surgery does not confer significant benefit compared with open surgery in the treatment of diverticula thoracic esophagus. PMID- 11852393 TI - [Malnutrition is an independent risk factor of early complications following percutaneous endoscopic gastrostomy]. AB - BACKGROUND: The relationship between malnutrition and complications occuring in hospitalized patients is debated. AIM: To assess prospectively the relationship between nutritional status and percutaneous endoscopic gastrostomy (PEG)-related complications and to determine the respective role of malnutrition and confounding variables on clinical outcome. METHODS: Ninety seven patients were followed up for 30 days after PEG insertion and complications were prospectively recorded. Patients were classified as malnourished when they met one of the following criteria: body mass index<20 kg/m(2) and/or weight loss > 5%. Multivariate logistic regression analysis was used to adjust for age, sex, diagnoses, functional status, antibiotic treatment and duration of hospital stay before PEG insertion. RESULTS: Sixty eight (70.1%) patients were malnourished. A significantly higher number of complications was seen in malnourished than in non malnourished patients (54.4% vs 31%; odds-ratio (OR): 2.65 [CI 95%: 1.06-6.66]; P=0.04). One-month mortality did not significantly differ between malnourished and non malnourished patients. Multivariate analysis showed that malnutrition (OR: 3.15 [1.21-8.24]; P=0.02) and > 10-day hospital stay (OR: 2.77 [1.13-6.82]; P=0.03) were significantly associated with early complications. CONCLUSION: These data suggest that malnutrition increases the risk of early complications after PEG insertion, independently of the underlying diseases and functional status of the patients. PMID- 11852394 TI - [Angioneurotic edema]. PMID- 11852395 TI - [Radiation-induced stricture of the papilla and the common bile duct: successful treatment with balloon dilation]. AB - Radiation-induced lesions of the bile ducts rarely occur and may be difficult to manage. We report the case of a 59-year old woman who developed radiation-induced stenosis of the papilla and the common bile duct 25 years after abdominal radiation therapy for abdominal non-Hodgkin's lymphoma. She presented with recurrent cholangitis and chronic cholestasis. Endoscopic results showed dilation of the intrahepatic bile ducts, radiation-induced inflammation and narrowing of the antrum and the duodenum, and stricture of the papilla and the last few millimeters of the common bile duct. The patient was treated with endoscopic balloon dilation. Forty-two months after endoscopic dilation, the patient remained asymptomatic with normal liver tests and no biliary dilation at ultrasound. PMID- 11852396 TI - [Colonic actinomycosis. Clinical, endoscopic and histological aspects in one patient]. AB - Colonic location of actinomycosis must be distinguished from infected colonic carcinoma and other colonic inflammatory pseudotumors. The diagnosis is often made by histological examination of the tumor. Initial treatment consists in surgery, followed by an antibiotic treatment. We report the case of a 32-year old woman with a right colonic actinomycosis. PMID- 11852398 TI - [Orbital metastasis of hepatocellular carcinoma]. PMID- 11852397 TI - [Tamoxifen for metastatic pancreatic adenocarcinoma: a complete response]. AB - Pancreatic adenocarcinoma tissue contains estrogen receptors and some pilot studies have suggested that tamoxifen could increase the survival of patients with unresectable pancreatic cancer. However data of comparative studies are conflicting. We report the case of a woman who presented with unresectable pancreatic head carcinoma with hepatic metastasis. The patient refused chemotherapy and was treated with tamoxifen 30 mg/day. A complete clinical remission of 50 months was observed. A further large-scale study may be required to assess the usefulness of tamoxifen in the second-line treatment of advanced pancreatic carcinoma. PMID- 11852399 TI - [Interobserver reproducibility of the Knodell score and the Metavir score in chronic viral hepatitis C. Results of 60 liver biopsies]. PMID- 11852400 TI - [Giant esophageal ulcer probably due to etidronate]. PMID- 11852401 TI - [Intestinal pseudo-obstruction with hypoganglionosis during acute pandysautomia]. PMID- 11852402 TI - [Van Hippel-Lindau disease presenting as recurrent acute pancreatitis]. PMID- 11852403 TI - [Acute pancreatitis after treatment by levofloxacin and methylprednisolone]. PMID- 11852404 TI - [Purtscher' ischemic retinopathy consecutive to an acute pancreatitis]. PMID- 11852405 TI - [Relapsing and metastatic evolution of a papillary cystic and solid tumor of the pancreas, twenty years after a first resection]. PMID- 11852406 TI - [Hepatitis B vaccination and the risk of multiple sclerosis: case-control studies]. PMID- 11852407 TI - [Search for microsatellite instability in colorectal cancer: an effective way to detect HNPCC?]. PMID- 11852408 TI - [Prophylactic gastrectomy in patients with deleterious E-cadherin gene mutation]. PMID- 11852409 TI - Prospective surveillance applying the national nosocomial infection surveillance methods in a Brazilian pediatric public hospital. AB - BACKGROUND: A hospital-wide, traditional prospective surveillance for nosocomial infections was commenced in 1992 in Centro Geral de Pediatria in Minas Gerais, Brazil, to describe the epidemiology of nosocomial infection in this pediatric hospital and to implement cross-infection prevention and control policies. METHODS: We performed a prospective cohort nosocomial infection surveillance of all patients receiving acute care according to the hospital-wide and intensive care unit components of the National Nosocomial Infections Surveillance System from January 1993 to December 1997 (14,892 discharges; 131,764 patient-days). The Centers for Disease Control and Prevention (Atlanta) 1988-definitions and the Brazilian Ministry of Health-Legislation 930 (1992) were used. RESULTS: The average overall nosocomial infection rate per 1000 patient-days was 8.9 in units 2 and 3 and 16.4 in the pediatric intensive care unit. Over time, the overall hospital infection rate decreased from 16.6 nosocomial infections per 1000 patient-days in 1993 to 7.0 in 1997 (P <.05). We believe this can be attributed to interventions and data reporting during the period. The five most frequent sites of infections were eye-ear-nose-throat (38%), skin (22%), pneumonia (12%), soft tissue (5%) and laboratory-confirmed bloodstream infection (4%). In the pediatric intensive care unit, the most frequent nosocomial infection sites were pneumonia related to mechanical ventilators (22%), with rates ranging from 0 to 42 per 1000 ventilator-days; and sepsis related to central lines (11%), with rates ranging from 0 to 32 per 1000 central line-days. CONCLUSIONS: Describing the epidemiology of nosocomial infections in this hospital enabled us to establish infection occurrence, distribution, and expected incidence, as well as to recognize trends and keep track of possible outbreaks. The knowledge acquired through this surveillance allowed us to target more specific and continuous quality improvement projects, to upgrade health care quality in pediatric public hospitals in Brazil, and to implement preventive strategies. Methods from the National Nosocomial Infections Surveillance System can be successfully applied in pediatric public hospitals in Brazil. PMID- 11852410 TI - Attitudes toward HIV-infected individuals and infection control practices among a group of dentists in Mexico City--a 1999 update of the 1992 survey. AB - BACKGROUND: The teaching of infection control (IC) was introduced at dental schools in Mexico during the 1990s. A 1992 survey indicated that some dentists had limited access to current IC standards. Deficient knowledge of bloodborne pathogens may influence dentists' attitudes about infected individuals and reduce compliance with IC recommendations. OBJECTIVE: To update the 1992 appraisal of attitudes about persons infected with HIV or the hepatitis B virus (HBV) and IC knowledge and practices in a nonrepresentative sample of dentists in Mexico City. METHOD: One hundred eighty dentists were interviewed in 1999 (response rate, 84.1%) with the same methods used in 1992. RESULTS: Seventy-nine percent of respondents perceived the risk of HIV infection as "considerable" to "very strong." The risk of HBV infection was considered higher than that of HIV. Only 32% of respondents had not been immunized against HBV. Reported use of personal protective equipment remained high. Dry heat was the preferred method for sterilization in 1992, but by 1999 it had been displaced by steam under pressure. Reported preference for more effective disinfectants was also evident overall. CONCLUSIONS: We found certain improvements in IC knowledge and practices between 1992 and 1999, and the results suggest targets for educational and regulatory efforts that are needed to promote better adherence to current IC standards. PMID- 11852411 TI - Estimation of the risk of bloodborne pathogens to health care workers after a needlestick injury in Taiwan. AB - OBJECTIVES: To estimate the number of health care workers (HCWs) in Taiwan at risk annually for contracting hepatitis B virus (HBV), hepatitis C virus (HCV), and HIV after a needlestick and sharps injury (NSI) with a used hollow-bore needle. METHODS: All patients hospitalized in 1 tertiary hospital between September 1997 and June 1998 had routine pathological work-ups. On the first day of the months of September 1997, December 1997, March 1998, and June 1998, 1805 samples of deidentified residual sera randomly sampled from 18,474 inpatients older than 6 years were serologically tested for antigens to HBV (HBsAg and HBeAg) and antibodies to HCV (anti-HCV) and HIV (anti-HIV) with enzyme-linked immunosorbent assay reagents. The frequency of NSIs with contaminated devices in HCWs from 16 public teaching hospitals between July 1996 and June 1997 and the serologic results were used to extrapolate the estimated annual rate of seroconversion in HCWs after an NSI. RESULTS: Of the 1805 samples tested, 16.7% were seropositive for HBsAg (of which 1.7% were positive for HBeAg), 12.7% were positive for anti-HCV, and 0.8% were positive for anti-HIV. Of the 7550 NSIs reported by 8645 HCWs, 66.7% involved a contaminated hollow-bore needle. From these data, 308 to 924 HCWs were estimated to be at risk for contracting HBV; 334 to 836 were at risk for contracting HCV; and, at the most, 2 were at risk for contracting HIV. The estimated annual number of contaminated NSIs sustained by 4 categories of HCWs ranged from 0.3 to 0.7, resulting in 543 nurses, 113 technicians, 80 physicians, and 66 supporting staff to be at risk annually of acquiring HBV infection. The numbers of HCWs estimated to be at risk of acquiring HCV were 596 nurses, 90 physicians, 84 technicians, and 30 supporting staff. The risk of acquiring HIV was low, with 1 nurse and possibly 1 other staff potentially exposed annually. CONCLUSIONS: Our estimates of the risk for seroconversion after an NSI have demonstrated that an occult risk can be formulated into a quantifiable risk. The number of susceptible HCWs at risk for seroconversion is as many as 1762 annually. With the number of nurses employed and the frequency with which they use sharps and sustain an NSI, 64.7% of all possible seroconversions will be in the nursing staff. This is a salient reminder of the importance of the introduction of early training in safe-needle-handling techniques before nurses enter their internship in countries where safety equipment, safety instructions, and staff vaccination programs are absent. PMID- 11852412 TI - Clinical and epidemiologic significance of coagulase-negative staphylococci bacteremia in a tertiary care university Israeli hospital. AB - OBJECTIVES: To characterize the clinical significance of coagulase-negative staphylococci (CNS) bacteremia. DESIGN: Prospective cohort study. SETTING: A 900 bed hospital in Haifa, Israel, from November 1996 to March 1997. RESULTS: Of 137 episodes of positive blood cultures for CNS, 41 (30%) were considered as true infection. Twenty-seven of 119 episodes associated with only 1 blood culture positive for CNS (23%) met the definition of infection as compared with 14 of 18 episodes (78%) associated with 2 or more blood cultures positive for CNS (P <.001). Methicillin resistance was significantly more frequent among Staphylococcus epidermidis isolates of episodes of true bacteremia than of episodes of contamination (15 of 22 [68%] vs. 11 of 33 [33%], respectively; P =.02). S hominis was isolated only in episodes considered as contamination (P =.01). It was estimated that CNS represents 24% of all nosocomial bloodstream pathogens. When CNS were isolated in the first 48 hours of hospitalization, an intravascular device was more frequently associated with episodes of true bacteremia than in those considered as contamination (7 of 7 [100%] vs. 10 of 57 [18%], respectively; P <.001). The mortality rate among patients with true CNS bacteremia was 16%. CONCLUSION: Some laboratory parameters may help identify episodes of true CNS bacteremia, which appears to be more common than previously considered. PMID- 11852413 TI - Nosocomial infections in a neonatal intensive care unit: incidence and risk factors. AB - BACKGROUND: Nosocomial infections (NIs) have become a matter of major concern in neonatal intensive care units (NICUs). The objectives of this study were to determine the incidence rate and the most frequent sites of infection in a Brazilian NICU from January 1999 to March 2000 and to study the risk factors for NIs. METHODS: A cohort study was carried out in which 225 neonates who remained at least 24 hours in the NICU were followed-up; neonates with NIs were identified, and the presence of risk factors was studied. Results were submitted to chi(2) distribution. RESULTS: The incidence rate and the incidence density rate were 50.7% and 62 infections per 1000 patient-days, respectively. In order of frequency, the sites of infection were: pneumonia (40.3%), primary bloodstream (16.7%), skin and soft tissue (14.9%), and meningitis (9.6%). The following risk factors were associated with NIs (P <.05): birth weight, gestational age, mechanical ventilation, total parenteral nutrition, umbilical catheter, use of antibiotics, and intubation in the delivery room. CONCLUSION: Risk factors were similar to those reported by other authors. However, incidence rates of infections in our NICU were much higher, possibly because of different methodologies and the adopted criteria for the classification of NIs. PMID- 11852414 TI - Surveillance of ventilator-associated pneumonia in very-low-birth-weight infants. AB - BACKGROUND: Surveillance of ventilator-associated pneumonia (VAP) is an essential part of quality patient care. Very-low-birth-weight (VLBW) infants, many with tracheal microbial colonization and bronchopulmonary dysplasia (BPD), comprise a difficult group in whom to make a diagnosis of pneumonia with the Centers for Disease Control and Prevention (CDC) criteria for infants younger than 1 year. OBJECTIVE: Our objective was to retrospectively compare VAP surveillance diagnoses made by the hospital infection control practitioner (ICP) with those made by a panel of experts with the same clinical and laboratory evidence and supportive radiologic data. A secondary objective was to compare radiologic diagnosis of pneumonia made by the general hospital radiologists, by the panel of experts, and by a pediatric radiologist from another hospital. STUDY POPULATION: Thirty-seven VLBW infants identified as at risk for VAP by the ICP on the basis of a positive bacterial tracheal culture and the application of CDC criteria for the definition of pneumonia were studied. METHODS: Clinical and laboratory evidence and routine radiologic reports made by the general radiologist were reviewed independently by a panel of experts composed of 3 experienced neonatologists. Chest x-rays from the day before, day of, and day after the surveillance date were reviewed separately by the 3 neonatologists and also by a pediatric radiologist. RESULTS: After inter-reader reliability was found satisfactory (kappa's coefficient, 0.47-0.75; P <.05), the panel of neonatologists determined that the 37 VLBW infants represented 4 distinct clinical categories. Group 1 comprised 12 airway-colonized infants, aged 14 to 30 days, who on the surveillance date, albeit intubated, were asymptomatic, not treated with antibiotics, and survived. Group 2 comprised 11 airway-colonized infants, aged 7 to 42 days, who presented with equivocal clinical, laboratory, or radiologic signs of VAP and survived. Group 3 comprised 7 airway-colonized infants, aged 14 to 21 days, who were acutely ill (3 died) and had clinical and laboratory evidence of nosocomial bloodstream infection (BSI) but no radiologic signs of pneumonia. Group 4 comprised 7 infants, aged 14 to 28 days, who were acutely ill (4 died) and had clinical and laboratory evidence of infection and radiologic changes consistent with VAP. Radiologic Findings: General radiologists, neonatologists, and the pediatric radiologist agreed that none of the asymptomatic airway-colonized infants (Group 1) had VAP. General radiologists reported signs suggestive of pneumonia in 8 of 11 infants (Group 2), a finding not corroborated by the others. Everybody agreed on the absence of radiologic pneumonia in 6 of 7 patients with nosocomial BSI (Group 3) and on the presence of signs consistent with pneumonia in the remaining 7 infants (Group 4). CONCLUSION: Surveillance diagnosis of VAP in VLBW infants is difficult because current CDC definitions are not specific for this population. Isolated positive tracheal culture alone does not distinguish between bacterial colonization and respiratory infection. Clinical and laboratory signs of VAP, mostly nonspecific, can be found in other conditions such as bronchopulmonary dysplasia and nosocomial BSI. Routine radiologic reports suggestive of pneumonia in airway-colonized infants without definitive clinical and laboratory evidence of infection could be misleading. To improve accuracy, surveillance diagnosis of VAP in special populations such as VLBW infants should be reformulated; meanwhile, ICPs should seek consultation with experienced clinicians for interpretation of data. PMID- 11852416 TI - Bacterial contamination during routine formula preparation. PMID- 11852415 TI - The effect of active surveillance for vancomycin-resistant enterococci in high risk units on vancomycin-resistant enterococci incidence hospital-wide. AB - BACKGROUND: Vancomycin-resistant enterococci (VRE) have become a major cause of nosocomial infections and are now endemic in many geographic areas. The aim of this study was to describe the effect of active surveillance for patients with VRE in high-risk units on the VRE incidence rate hospital-wide. METHODS: We determined 4 time periods based on the intervention of active surveillance: preactive surveillance (period 1), active surveillance (period 2), no active surveillance (period 3), and reinstutition of active surveillance (period 4). VRE incidence rates based on first clinical culture for VRE per 10,000 patient days for each of these periods and incidence rate ratios were then calculated. RESULTS: Active surveillance in high-risk units was associated with a significant reduction in VRE incidence hospital-wide in 2 of the 3 comparisons made. The incidence rate ratio when comparing the first period of active surveillance (period 2) to the preactive surveillance period (period 1) was 0.63 (95% CI, 0.38 1.1); it was 0.36 (95% CI, 0.23-0.55) when comparing the first period of active surveillance (period 2) to the subsequent period (period 3) and 0.68 (95% CI, 0.54-0.85) when comparing the second period of active surveillance (period 4) to the prior period without active surveillance periods. CONCLUSIONS: Active surveillance culturing for VRE in the high risk-units prevented further VRE transmission, as evidenced by a significant increase in hospital-wide incidence rates when active surveillance was discontinued and a significant decrease in incidence rates when it was restarted. PMID- 11852417 TI - Bioterrorism preparedness: a survey of Nebraska health care institutions. AB - In March 2001, a 6-question survey was mailed to all hospitals and long-term care facilities in Nebraska to assess preparedness for bioterrorism. Only half of the respondents at that time believed that bioterrorism was something their community was likely to experience. We found that most facilities (98%) believed that they were unprepared for a bioterrorism event, and many did not know whom to contact in the event of such an emergency. We concluded from the results of the survey that the greatest needs to facilitate preparation were policies and procedures, identification of contacts, medications, protective equipment, laboratory support, and communication. PMID- 11852418 TI - The implementation of a surgical antibiotic prophylaxis program: the pivotal contribution of the hospital pharmacy. AB - BACKGROUND: Although surgical site infection rates have decreased with the prophylactic use of antibiotics, the inappropriateness of surgical antibiotic prophylaxis is still a worldwide problem. Various strategies have been used to address this problem. This study describes the implementation of a perioperative antibiotic prophylaxis protocol that emphasizes the contribution of the pharmacist. METHODS: A descriptive study design was used to evaluate the impact of the protocol on the appropriateness of prophylaxis in a private university hospital. The surgical antibiotic prophylaxis of all surgeries was evaluated for 1 month before and 1 month after the implementation of the protocol. RESULTS: The appropriateness of the indication for prophylaxis rose from 56.4% to 100% and that of the postoperative maintenance prophylactic antibiotics rose from 21.9% to 95.7%. The cost of the perioperative antibiotic prophylaxis per surgery decreased 40.5%. CONCLUSION: The implementation of a cost-effective perioperative antibiotic prophylaxis protocol was the result of a multidisciplinary effort. The hospital pharmacist participated in education activities as part of the discussion groups on the perioperative antibiotic prophylaxis protocol that involved all participants and in managerial actions that optimized the process of ordering, dispensing, administering, and documenting the perioperative antibiotic prophylaxis. PMID- 11852419 TI - Infection control in Saudi Arabia: meeting the challenge. AB - Hospital-acquired infection poses significant clinical and economic burden worldwide. In the Kingdom of Saudi Arabia, infection control is a young, rapidly growing specialty. An infrastructure to expedite the growth of this important discipline is fast being established. The kingdom faces unique challenges when addressing infection control, which are the subject of this review. Much of the policy-making in domestic infection control is driven by the preventive medicine concerns of the annual pilgrimage (Hajj) to Mecca, which are unparalleled. The Saudi Ministry of Health acts to contain and control public health risks at this gathering of 2 million. Infectious hazards at the Hajj include meningococcal meningitis, respiratory tract infections, bloodborne diseases, and zoonotic diseases, all of which have international ramifications as pilgrimaging Muslims return home. In the wake of the extraordinary pace of modernization in Saudi Arabia, deficiencies in infection control remain, which are slowly being redressed. This review examines the anatomy of infection control and its evolution in the kingdom. Future goals and infection control policy-making are given particular emphasis. Saudi Arabia seeks increasing international partnership in the area of infection control and preventive medicine. The Saudi health care system was formed on the basis of Western models to resounding success. Saudi Arabia is now in a position to provide experience and knowledge in return. International dialogue in the infection control arena is of mutual value. Important public health progress is afoot in this young kingdom, and these advances translate both regionally and on the international platform. PMID- 11852420 TI - Limited surveillance in the endoscopic setting: has its time arrived? PMID- 11852421 TI - Descriptive study of nosocomial infections in a short-term inpatient behavioral health setting. PMID- 11852422 TI - Improving benchmarks for surveillance by defining types of pediatric intensive care units. PMID- 11852423 TI - Dual action of nitric oxide on purely isolated retinal ganglion cells. AB - PURPOSE: The role of nitric oxide (NO) in the survival of retinal ganglion cells (RGCs) was investigated. METHODS: RGCs were purely isolated from postnatal Sprague-Dawley rats by 2-step panning and were cultured in chemically defined serum free medium. An NO releaser, S-nitroso-N-acetylpenicillamine (SNAP: 500 microM, 250 microM, 100 microM, 10 microM, 1 microM, 100 nM, and 10 nM), an NO scavenger, 2-(4-carboxyphenyl)-4,4,5,5 tetramethylimidazoline-1-oxyl-3-oxide potassium salt (c-PTIO: 100 microM, 33 microM, 10 microM, 1 microM), mixture of 100 microM SNAP and 33 microM c-PTIO, N(G)-nitro-L-arginine methyl ester (L-NAME: 10 mM, 5 mM, 500 microM, 100 microM or 10 microM), or their vehicles were added to the medium of pure RGC culture for 48 hr. Survival rates of small and large RGCs were determined separately by flow cytometry. RESULTS: At > or = 100 microM, SNAP significantly reduced RGC survival in a concentration dependent manner. At < or = 41 microM, SNAP significantly increased survival, particularly of large RGCs. c-PTIO and L-NAME reduced the survival rates concentration-dependently. A mixture of 100 microM SNAP and 33 microM c-PTIO significantly improved RGC survival compared with when they were added on their own. CONCLUSIONS: These results indicate that NO exhibits neuroprotective and neurotoxic actions on RGCs and that low concentrations of NO may be beneficial for the survival of neonatal RGCs in vitro. PMID- 11852424 TI - Corrosion casts and scanning electron microscopy of choroidal vasculature in rats with inherited hypercholesterolemia. AB - PURPOSE: The purpose of the present study is to determine the specific morphological changes in choroidal vasculature due to hypercholesterolemia. METHODS: The choroidal vessels of rats with inherited hypercholesterolemia (RICO) were examined by the corrosion cast technique and electron microscopy. RESULTS: In 18-month-old RICO rats, corrosion casts and scanning electron microscopy showed severe tortuosity, caliber irregularity and generalized dilatation of the large choroidal arteries and veins. The branch arterioles and draining venules were also tortuous and dilated. The ampullae of the vortex veins were wide, but no arteriovenous anastomoses, crossing defects or obstruction were seen. However, there were many venular collateral channels and venulo-venular anastomoses. The choriocapillaries appeared normal. Transmission electron microscopy of the choroidal vessels showed hypertrophy of smooth muscle cells and irregular thickening of the basement membrane. The RPE and Bruch's membrane were intact. CONCLUSION: These findings indicate that the choroidal vascular changes are probably related to hypercholesterolemia. The results of the present study will be useful in the interpretation of the changes seen in hypertensive, diabetic and aged choroids. PMID- 11852425 TI - Circadian change of adenylate cyclase activity in rabbit ciliary processes. AB - PURPOSE: To characterize in vivo changes of adenylate cyclase activity in rabbit ciliary processes during the circadian cycle. METHODS: After removal of vitreous, lens, retina and choroid from freshly enucleated rabbit eyes, the anterior segment and attached sclera was chilled in cold buffer containing the non selective phosphodiesterase (PDE) inhibitor, 3-isobutyl-1-methylxanthine (IBMX). Then ciliary processes were excised and homogenized in ice cold trichloro-acetic acid (TCA) 2.5 min after IBMX treatment. Increased cyclic AMP in response to PDE inhibition was measured in ciliary processes at six times during the circadian cycle, after preganglionic section of the cervical sympathetic trunk (decentralization, DX), and after topical instillation of hydroxyamphetamine, timolol, brimonidine, rauwolscine or a soluble derivative of forskolin. RESULTS: The increase of rabbit ciliary process cyclic AMP levels in response to PDE inhibition with IBMX was enhanced at night. Much of the enhanced nocturnal increase persisted in constant dark and was blunted by DX. Topical instillation of hydroxyamphetamine enhanced the increase during the day; whereas, timolol, a beta-adrenergic antagonist, or brimonidine, an alpha2-adrenergic agonist, blunted the IBMX induced increase. Neither instillation of rauwolscine, an alpha2 adrenergic antagonist, nor the soluble forskolin derivative enhanced the increase of cyclic AMP after IBMX. CONCLUSIONS: The technique reported here can be used to estimate ciliary process adenylate cyclase activity in vivo. There is a circadian rhythm of adenylate cyclase activity in rabbit ciliary processes that is driven in part by ocular sympathetic input and stimulation of beta-adrenergic receptors. PMID- 11852426 TI - Development and survival of tyrosine hydroxylase containing neurons in RCS rat retinae. AB - AIM: Dopamine serves a variety of functions in the retina. Abnormalities of the retinal dopaminergic system have been described in the Royal College of Surgeons (RCS) rat as well as other models of retinal degeneration. Dopamine has been implicated in several retinal dysfunctions of retinitis pigmentosa. Dopaminergic amacrine cells respond to light by increasing their tyrosine hydroxylase (TH) activity and the rate of dopamine turnover. This study has, therefore, examined the ontogenesis of TH containing cells in the RCS rat retina to assess whether progressive photoreceptor degeneration affects the development or survival of TH containing cells in any way. METHODS: TH immunoreactivity in developing dystrophic RCS rat retinae (postnatal day (PN) 0, 3, 6, 14, 18, 26, 32, 56, 85, 91, 12 month and 15 month) and normal retina (PN day 0, 6, 14, 19, 26, 30, 33, 54 and adults) was compared. RESULTS: TH immunoreactivity in dystrophic retina closely resembled that in normal retina. In both groups, very faintly immunoreactive cells were detected in the proximal retina at PN 0. Immunoreactivity increased until PN 14, when faintly immunoreactive interplexiform (IP) fibers and fibers in the outer plexiform layer could be observed. In both groups, the IP connections reached their mature level of development at about PN 30. Thus the developmental expression of TH immunoreactive cells resembled that of non-dystrophic retina in both chronology as well as types of cells. These cells survived even in the advanced stages of degeneration. CONCLUSIONS: The results suggest that the abnormalities in the dopaminergic system of the RCS retinae are not associated with abnormal ontogeny or survival of TH synthesizing cells. PMID- 11852428 TI - Photodynamic therapy with indocyanine green for occult subfoveal choroidal neovascularization caused by age-related macular degeneration. AB - PURPOSE: To report two cases of occult subfoveal choroidal neovascularization (CNV) due to age-related macular degeneration (AMD) that were successfully treated utilizing large spot, low irradiance 810-nm light and intravenous injection of small-volume, high-concentration indocyanine green (ICG) bolus, termed ICG-mediated photodynamic therapy (i-PDT). METHODS: Case report. Two patients with occult subfoveal CNV (two eyes) due to AMD were treated with i-PDT and were prospectively followed with fluorescein and indocyanine green angiography as well as optical coherence tomography (OCT). RESULTS: Both patients presented a 3-line increase in visual acuity by twelve months of follow-up. A significant regression of the CNV was demonstrated by fluorescein angiography within one week following treatment. OCT revealed diminishing of the subretinal fluid. There were no complications related to the procedure. CONCLUSION: i-PDT induced CNV regression and improved the visual acuity in these particular cases, suggesting a new and promising way to treat neovascular AMD. The partial anatomical restoration associated to the improvement of the visual function observed in both patients suggests that a selective therapeutic effect was achieved. Further studies are required not only to assess the clinical value of this treatment but also to better understand the major mechanism of action involving this novel treatment modality. PMID- 11852427 TI - Binding characteristics of ET receptors in retinal pericytes and effects of high glucose incubation. AB - PURPOSE: Autoregulation and control of blood flow within the retina is dependent on vascular endothelial release of vasoconstrictors and dilators. The effector cells are the pericytes and vascular smooth muscle cells of the retinal microvascular bed. Many studies show that the responses of these effector cells are affected by diabetes or artificially elevated glucose. In particular, the contractile response of pericytes to endothelin-1 (ET-1) is markedly reduced. ET 1 and related isopeptides exert their effects by binding and activating specific cell surface receptors (ETRs). Pericytes express two subtypes of ETRs, ET(A) and ET(B). The purpose of this study was to assess the dynamics of [(125)I]ET-1 binding to ET(A) and ET(B) receptors when cultured in normal and high glucose concentrations. METHODS: Bovine retinal pericytes (BRP) were exposed to normal (5 mM) or high glucose (25 mM) concentrations for up to 14 days. Displacement of binding and competitive inhibition of binding was carried out in perictye membranes. mRNA expression for ETR subtypes was examined using northern blotting. The expression of G protein alpha subunits (Galpha(s) and Galpha( q)) was detected by western blotting. RESULTS: Highly specific binding of [125I]ET-1 was seen with BRP membranes. In competition assays, ET-3 was found to displace [125I]ET-1 binding to BRP membranes in a biphasic manner, indicating the presence of two classes of binding sites. The presence of both ETR subtypes was supported by competition experiments with selective ligands. The expression of Galpha(s) and Galpha(q) subunits decreased in pericytes incubated with 25 mM glucose. Conclusions. No significant differences in binding parameters were evident after culture in high glucose concentrations. However northern blot analysis which confirmed the presence of ET(B) receptor mRNA did identify an increase in the expression of this receptor. PMID- 11852429 TI - Retinal glial cell immunoreactivity and neuronal cell changes in rats with STZ induced diabetes. AB - PURPOSE: To study whether diabetes could influence glial cells, retinal neurons, and pigment epithelial cells and if so, to evaluate whether any changes could be influenced by aminoguanidine (AG) or probucol (PB). METHODS: Streptozocin (STZ) induced diabetic male Wistar rats and age-matched control rats were fed a normal diet, addition of AG in the drinking water (0.5 g/l for diabetic and 1.0 g/l for control rats) or PB in the pellets (1 % w/w) for one or six months. Paraffin embedded retinal sections were incubated in the primary antibodies GFAP, calbindin, RPE65, and Hu, for glial, horizontal, pigment epithelial, and ganglion cells, respectively, and in fluorescent secondary antibodies. RESULTS: One month after STZ injection, GFAP immunoreactivity was sparse, but after six months it was prominent in glial cells in 5/5 diabetic and 1/7 control retinas (p = 0.015). Neither AG, nor PB influenced this immunoreactivity. Numbers of retinal pigment epithelial cells and cells in the ganglion cell layer, were similar at one and six months of diabetes. By time, the number of horizontal cells decreased (p < 0.001) and branching and numbers of their terminals were reduced (p < 0.001). CONCLUSION: Diabetes for six months resulted in increased glial cell immunoreactivity, and by age, horizontal cell numbers and branching of their terminals decreased, morphological patterns that were unaffected by AG or PB. The numbers of retinal pigment epithelial cells and cells in the ganglion cell layer were unaffected both by age and diabetes. PMID- 11852430 TI - Cytokine modulation of costimulatory molecules on human fetal retinal pigment epithelial cells. AB - PURPOSE: Experiments were performed to evaluate the effect of various pro- and anti-inflammatory cytokines on the human fetal retinal pigment epithelium's (HFRPE) expression of major histocompatibility complex (MHC) and costimulatory molecules. METHODS: Pure cultures of HFRPE cells were isolated. HFRPE cells were incubated in the presence of Interferon-gamma (IFN-gamma), IFN-beta, Tumor Necrosis Factor-alpha (TNF-alpha), Interleukin-1beta (IL-1beta), Tumor Growth Factor-beta (TGF-beta), and a combination of IFN-gamma and TGF-beta (pre incubation and simultaneously incubated). The expression of MHC class I and class II, Intercellular cell adhesion molecule (ICAM-1), B7-1 (CD80), and B7-2 (CD86) molecules was quantitatively analyzed by flow cytometry. RESULTS: The cultured HFRPE cells expressed high levels of MHC class I and low levels of MHC class II and ICAM-1 molecules. After culture with the above mentioned cytokines, IFN-gamma up-regulated the HFRPE's expression of MHC class II and ICAM-1. IFN-beta and IL beta1 only up-regulated the expression of ICAM-1. TGF-beta was unable to suppress the up-regulatory effect of IFN-gamma in HFRPE cells (pre-incubated and simultaneously incubated). The other cytokines did not have any significant effect on HFRPE's expression of MHC I and II or the selected costimulatory molecules. CONCLUSIONS: Our findings indicate that TGF-beta cannot suppress up regulating effects of IFNgamma- on HFRPE's expression of MHC and costimulatory molecules. Overall, the weak or lack of expression of costimulatory molecules after stimulation with various cytokines further confirms that HFRPE cells are weak antigen presenting cells. PMID- 11852431 TI - Effect of caffeic acid phenethyl ester on corneal neovascularization in rats. AB - PURPOSE: Caffeic acid phenethyl ester (CAPE), a biologically active component of propolis from honeybee hives, has potent antiinflammatory and antioxidant properties. We aimed to evaluate the ability of topically applied CAPE in comparison with known steroidal (dexamethasone sodium phosphate) and nonsteroidal (indomethacin) topical agents to reduce corneal neovascularization (CNV) induced by silver nitrate cauterization in rats. METHODS: Following silver nitrate cauterization on both eyes, male rats were randomly assigned to the study and control groups, each consisting of ten rats. The inhibitory effects of the test drugs against a placebo (isotonic saline) on CNV were tested and compared to each other using a previously described method in which extent of neovascularization and burn stimulus intensity were scored by a masked examiner. Briefly, burn stimulus intensity was scored from 0 to +3 according to the height of blister from corneal surface, and extent of neovascularization was recorded from 0 to +6 according to the distance from limbus to the end point of CNV toward the central corneal burn. Results. The mean burn stimulus score were not different among the groups (P = 0.807). Percent inhibition of CNV compared to the placebo control and its significance were 31.5 %, P = 0.011 for indomethacin; 56 %, P < 0.001 for dexamethasone; and 52 %, P < 0.001 for CAPE. Dexamethasone was significantly (P < 0.05) more effective than indomethacin in inhibition of neovascular growth. CAPE was found to be superior (P < 0.05) to indomethacin and almost as effective as (P > 0.05) dexamethasone in reducing CNV. Conclusion. Topically applied CAPE was demonstrated to have an inhibitory effect, comparable to that of topical dexamethasone, on CNV in this rat model. Antiinflammatory and antioxidant properties of CAPE may contribute to its suppression on CNV. PMID- 11852432 TI - Short-term effect of beta-adrenoreceptor blocking agents on ocular blood flow. AB - PURPOSE: In this study the acute effect of the topically-delivered non-selective beta-blockers timolol and carteolol, and the selective beta-blocker betaxolol, were evaluated with respect to ocular blood flow, intraocular pressure (IOP) and vessel resistance in rabbits' eyes. METHODS: In a double masked randomized design, one eye of each subject (n = 9) received two drops of 0.5 % timolol or 2 % cartelol or 0.5 % betaxolol ophthalmic solution and a separate group of nine rabbits received two drops of placebo consisting of physiological saline in both eyes to serve as control. Using hydrogen clearance method, ciliary body blood flow (CiBF), choroidal blood flow (CBF), and retinal blood flow (RBF) were measured. IOP and systemic mean arterial pressure (MAP) of each subject were measured under same condition before and after the administration of respective drugs to calculate the ocular perfusion pressure (OPP) and vessel resistance. RESULTS: In timolol- and carteolol-treated eyes significant reduction was observed in IOP (p < 0.01), CiBF (p < 0.01), CBF (p < 0.01) and RBF (p < 0.01) compared to control eyes. However, in betaxolol-treated eyes a marginal reduction in IOP was observed accompanied by significant increase in CiBF (p < 0.01) and RBF (p < 0.05). The non-selective beta-blocker-treated eyes tended to have increased vessel resistance, whereas, selective beta-blocker-treated eyes tended to have decreased vessel resistance. CONCLUSIONS: Our current results comparing non-selective and selective beta-blockers suggest that the selective beta-blocker betaxolol may be more appropriate for maintenance of retinal blood flow in situations with low perfusion. Currently the mechanism for regulation of IOP is unclear; however, the findings from this study indicate that decreased CiBF may contribute to reduction in IOP. PMID- 11852435 TI - Alopex-B: a new, simpler, but yet faster version of the alopex training algorithm. AB - Experimenting with some changes and simplifications to the Alopex algorithm, we obtained a new faster version (Alopex-B), that also shows lower failure rates on training attempts. Like Alopex, our version is network-architecture independent, does not require error or transfer functions to be differentiable, has a high potential for parallelism, and is stochastic (which helps avoid local minima), but unlike Alopex it follows no annealing scheme, and uses less parameters which makes it simpler to implement and to use. PMID- 11852433 TI - Involvement of inducible nitric oxide synthase in cataract formation in Shumiya cataract rat (SCR). AB - PURPOSE: Our previous results showed that the oral administration of aminoguanidine (AG), an inhibitor of inducible nitric oxide synthase (iNOS), strongly suppresses lens opacification in Shumiya cataract rat (SCR). Therefore, we examine whether iNOS is upregulated and involved in cataract formation in SCR. METHODS: The expressions of iNOS mRNA and iNOS protein in SCR lenses were examined by RT-PCR and Western blotting, respectively. Calpain-mediated alphaB crystallin proteolysis was analyzed by Western blotting using antibody specific to the calpain-generated fragment of alphaB-crystallin. Lens opacification was analyzed using computerized image analysis software connected to the Anterior Eye Segment Analysis System (EAS-1000, Nidek). Calcium contents in lenses were measured by atomic absorption spectrophotometry. RESULTS: High levels of iNOS mRNA and iNOS protein are expressed in cataractous lenses compared with normal lenses. The increases in their expression are markedly suppressed by the oral administration of AG, which acts to prevent lens opacification. The induction of iNOS protein is observed before the elevation in calcium content and the acceleration of calpain-mediated proteolysis, both of which are closely related to the development of lens opacification. CONCLUSIONS: These findings strongly suggest that iNOS is involved in cataract formation in SCR. The induction of iNOS occurs prior to the elevation of calcium content and its induction is inhibited by AG-treatment. Considering our previous result that the elevation of calcium content is also prevented by AG-treatment, it is conceivable that upregulation of iNOS causes calcium influx into lens cells and the subsequent activation of calpain. PMID- 11852436 TI - Improving ARTMAP learning through variable vigilance. AB - This paper presents a mechanism to vary the vigilance parameter in the RePART fuzzy neural network. This mechanism helps to smooth out the problem of category proliferation which affects ARTMAP-based networks. Empirical experiments show that the use of variable vigilance improves the performance of the RePART model while, at the same time, requiring a less complex structure. PMID- 11852437 TI - Automatic determination of radial basis functions: an immunity-based approach. AB - The appropriate operation of a radial basis function (RBF) neural network depends mainly upon an adequate choice of the parameters of its basis functions. The simplest approach to train an RBF network is to assume fixed radial basis functions defining the activation of the hidden units. Once the RBF parameters are fixed, the optimal set of output weights can be determined straightforwardly by using a linear least squares algorithm, which generally means reduction in the learning time as compared to the determination of all RBF network parameters using supervised learning. The main drawback of this strategy is the requirement of an efficient algorithm to determine the number, position, and dispersion of the RBFs. The approach proposed here is inspired by models derived from the vertebrate immune system, that will be shown to perform unsupervised cluster analysis. The algorithm is introduced and its performance is compared to that of the random, k-means center selection procedures and other results from the literature. By automatically defining the number of RBF centers, their positions and dispersions, the proposed method leads to parsimonious solutions. Simulation results are reported concerning regression and classification problems. PMID- 11852438 TI - Recruitment learning of boolean functions in sparse random networks. AB - This work presents a new class of neural network models constrained by biological levels of sparsity and weight-precision, and employing only local weight updates. Concept learning is accomplished through the rapid recruitment of existing network knowledge - complex knowledge being realised as a combination of existing basis concepts. Prior network knowledge is here obtained through the random generation of feedforward networks, with the resulting concept library tailored through distributional bias to suit a particular target class. Learning is exclusively local - through supervised Hebbian and Winnow updates - avoiding the necessity for backpropagation of error and allowing remarkably rapid learning. The approach is demonstrated upon concepts of varying difficulty, culminating in the well-known Monks and LED benchmark problems. PMID- 11852439 TI - A biologically inspired neural network for dynamic programming. AB - An artificial neural network with a two-layer feedback topology and generalized recurrent neurons, for solving nonlinear discrete dynamic optimization problems, is developed. A direct method to assign the weights of neural networks is presented. The method is based on Bellmann's Optimality Principle and on the interchange of information which occurs during the synaptic chemical processing among neurons. The neural network based algorithm is an advantageous approach for dynamic programming due to the inherent parallelism of the neural networks; further it reduces the severity of computational problems that can occur in methods like conventional methods. Some illustrative application examples are presented to show how this approach works out including the shortest path and fuzzy decision making problems. PMID- 11852440 TI - A fast and convergent stochastic MLP learning algorithm. AB - We propose a stochastic learning algorithm for multilayer perceptrons of linear threshold function units, which theoretically converges with probability one and experimentally exhibits 100% convergence rate and remarkable speed on parity and classification problems with typical generalization accuracy. For learning the n bit parity function with n hidden units, the algorithm converged on all the trials we tested (n=2 to 12) after 5.8 x 4.1(n) presentations for 0.23 x 4.0(n-6) seconds on a 533MHz Alpha 21164A chip on average, which is five to ten times faster than Levenberg-Marquardt algorithm with restarts. For a medium size classification problem known as Thyroid in UCI repository, the algorithm is faster in speed and comparative in generalization accuracy than the standard backpropagation and Levenberg-Marquardt algorithms. PMID- 11852441 TI - Estimation of hourly average solar radiation on tilted surface via ANNs. AB - An innovative approach is proposed for the estimation of hourly average solar radiation on tilted surface. The proposed approach, which is based on artificial neural networks and problem decomposition, demonstrates the following characteristics: Accuracy. Superior estimation is attained over both conventional approaches and theoretical models. Simplicity and efficiency. A small training set is employed and the training/test patterns involve few easily obtainable parameters. Generalization capability and robustness to noise. Apart from being of interest in meteorology, the accurate and efficient estimation of hourly average solar radiation on tilted surface is especially important in solar energy applications. PMID- 11852442 TI - Stability of dead zone bidirectional associative memory neural networks involving time delays. AB - A mathematical model describing the dynamical interactions of bidirectional associative memory networks involving transmission delays is considered. The influence of a dead zone or a zone of noactivation on the global stability is investigated and various easily verifiable sets of sufficient conditions are established. The asymptotic nature of solutions when the given system of equations does not possess an equilibrium pattern is discussed. PMID- 11852443 TI - Hierarchical genetic algorithm for near optimal feedforward neural network design. AB - In this paper, we propose a genetic algorithm based design procedure for a multi layer feed forward neural network. A hierarchical genetic algorithm is used to evolve both the neural networks topology and weighting parameters. Compared with traditional genetic algorithm based designs for neural networks, the hierarchical approach addresses several deficiencies, including a feasibility check highlighted in literature. A multi objective cost function is used herein to optimize the performance and topology of the evolved neural network simultaneously. In the prediction of Mackey Glass chaotic time series, the networks designed by the proposed approach prove to be competitive, or even superior, to traditional learning algorithms for the multi layer Perceptron networks and radial basis function networks. Based upon the chosen cost function, a linear weight combination decision making approach has been applied to derive an approximated Pareto optimal solution set. Therefore, designing a set of neural networks can be considered as solving a two objective optimization problem. PMID- 11852444 TI - New learning automata based algorithms for adaptation of backpropagation algorithm parameters. AB - One popular learning algorithm for feedforward neural networks is the backpropagation (BP) algorithm which includes parameters, learning rate (eta), momentum factor (alpha) and steepness parameter (lambda). The appropriate selections of these parameters have large effects on the convergence of the algorithm. Many techniques that adaptively adjust these parameters have been developed to increase speed of convergence. In this paper, we shall present several classes of learning automata based solutions to the problem of adaptation of BP algorithm parameters. By interconnection of learning automata to the feedforward neural networks, we use learning automata scheme for adjusting the parameters eta, alpha, and lambda based on the observation of random response of the neural networks. One of the important aspects of the proposed schemes is its ability to escape from local minima with high possibility during the training period. The feasibility of proposed methods is shown through simulations on several problems. PMID- 11852445 TI - Feature decomposition architectures for neural networks: algorithms, error bounds, and applications. AB - In recent years, systems consisting of multiple modular neural networks have attracted substantial interest in the neural networks community because of various advantages they offer over a single large monolithic network. In this paper, we propose two basic feature decomposition models (namely, parallel model and tandem model) in which each of the neural network modules processes a disjoint subset of the input features. A novel feature decomposition algorithm is introduced to partition the input space into disjoint subsets solely based on the available training data. Under certain assumptions, the approximation error due to decomposition can be proved to be bounded by any desired small value over a compact set. Finally, the performance of feature decomposition networks is compared with that of a monolithic network in real world bench mark pattern recognition and modeling problems. PMID- 11852446 TI - [Emergencies evolving from local anesthesia]. AB - Local anesthesia is without doubt the most frequently used drug in dentistry and in medicine. In spite of records of safety set by using these drugs, there is evidence to adverse reactions ranging from 2.5%-11%. Most of the reactions originate from the autonomic system. A recent, well-planned study indicates that adverse reactions are highly correlated to the medical status of the patient: the higher the medical risk, the greater the chance to experience an adverse reaction. This study also found that adverse reactions highly correlated to the concentration of adrenalin. Another recent study found a direct relationship between adverse reactions and the level of anxiety experienced by the patient and to the dental procedure. Most of the reactions in this study occurred either immediately at injection time and within 2 hours following the injection. Since the beginning of last century, vasoconstrictors have been added to local anesthesia solutions in order to reduce toxicity and prologue activity of the LA. However, today it is commonly agreed that this addition to local anesthesia should not be administered to cardiac patients especially those suffering from refractory dysrhythmias, angina pectoris, post myocardial infarction (6 months) and uncontrolled hypertension. Other contraindications to vasoconstrictors are endocrine disorders such as hyperthyroidism, hyperfunction of the medullary adrenal (pheochromocytoma) and uncontrolled diabetes mellitus. Cross reactivity of local anesthetic solutions can occur with MAO inhibitors, non specific beta adrenergic blockers, tricyclic antidepressants, phenothiazides and cocaine abusers. Noradrenaline added to local anesthetics as a vasoconstrictor has been described as a trigger to a great increase in blood pressure and therefore has been forbidden for use in many countries. This paper describes 4 cases of severe complications following the injections of local anesthesia of which three ended in fatality. PMID- 11852447 TI - [Syncope in the dental environment]. AB - Syncope or Fainting is, by far, the most common emergency situation in the dental practice. Syncope is defined as an abrupt, transient, short term loss of consciousness and postural tone, followed by spontaneous and complete recovery. The pathophysiology of syncope consists of a sudden cessation or decrease in cerebral perfusion. Differential diagnosis of these medical conditions is of paramount importance in uncovering unrecognized systemic diseases. The dental team plays an important role in the process of establishing the correct diagnosis by its ability to recognize and document all the clinical symptoms and signs evident at the time of fainting. The dental surgeon is expected to be familiar with the various etiologies of syncope and should be able to differentiate between them. This article provides the essentials of the diagnostic procedure and an approach to the evaluation of the unconscious patient. PMID- 11852448 TI - [Respiratory distress]. AB - Dental treatment is usually conducted in the oral cavity and in very close proximity to the upper respiratory airway. The possibility of unintentionally compromising this airway is high in the dental environment. The accumulation of fluid (water or blood) near to the upper respiratory airway or the loosening of teeth fragmentations and fallen dental instruments can occur. Also, some of the drugs prescribed in the dental practice are central nervous system depressants and some are direct respiratory drive depressors. For this reason, awareness of the respiratory status of the dental patient is of paramount importance. This article focuses on several of the more common causes of respiratory distress, including airway obstruction, hyperventilation, asthma, bronchospasm, pulmonary edema, pulmonary embolism and cardiac insufficiency. The common denominator to all these conditions described here is that in most instances the patient is conscious. Therefore, on the one hand, valuable information can be retrieved from the patient making diagnosis easier than when the patient is unconscious. On the other hand, the conscious patient is under extreme apprehension and stress under such situations. Respiratory depression which occurs during conscious sedation or following narcotic analgesic medication will not be dealt with in this article. Advanced pain and anxiety control techniques such as conscious sedation and general anesthesia should be confined only to operators who undergo special extended training. PMID- 11852449 TI - [Chest pains in the dental environment]. AB - Chest pain does not necessarily indicate cardiac disease. The most common causes of acute chest pain encountered in dental situations include hyperventilation, pulmonary embolism, angina pectoris and myocardial infarction. Stress and fear often cause rapid breathing or hyperventilation. This usually occurs in young adults and although the hyperventilating patient often complains of chest pain, this is rarely a manifestation of cardiac disease. Pulmonary embolism usually indicates the occlusion of a pulmonary artery causing severe chest pain. The primary clinical manifestation of angina pectoris is chest pain. Although most instances of anginal pain are easily terminated, the dentist must always consider the possibility that the supposed anginal attack is actually a sign of acute myocardial infarction (AMI). AMI is a clinical syndrome caused by a deficient coronary arterial blood supply to a region of myocardium that results in cellular death. There is a high incidence of mortality among AMI with death often occurring within 2 hours of the onset of signs and symptoms. The initial clinical manifestations of all types of chest pain can be similar. Therefore the dentist must develop proficiency in constituting a differential diagnosis and an efficient management protocol. As in most medical situations prevention is the most powerful tool. However, if chest pains do occur, measures such as airway management, oxygen supplementation, coronary artery dilation, analgesis and in extreme cases, cardiopulmonary resuscitation and evacuation to the emergency room, may be necessary. PMID- 11852450 TI - Medical emergencies in dentistry. PMID- 11852451 TI - [Allergy-related emergencies]. AB - Allergic reactions can develop to any of the drugs or materials commonly used in dentistry. They exhibit a broad range of clinical signs and symptoms ranging from mild, delayed reactions to immediate and life-threatening reactions developing within seconds. Allergies usually manifest themselves in reactions that are related to histamine release in one of three ways: skin reactions, respiratory problems and anaphylaxis. Anaphylaxis is the most critical allergic reaction in the dental environment. Measures such as airway management, oxygen supplementation, antihistamine, adrenaline and corticosteroid medication, cardiopulmonary resuscitation and evacuation to the emergency room, may be necessary. PMID- 11852452 TI - [The potential dangers of endocrinal disorders]. AB - The symptoms of most endocrine system diseases are usually clearly recognizable and most of the times are accompanied by a rich medical history. Many general practitioners are reluctant to treat such cases and prefer to refer these patients to specialists who are trained in management of the medically compromised thus increasing the chances of dental treatment without complications. However, sometimes endocrinal diseases develop slowly and their clinical manifestations are hidden or subclinical in nature. In these cases, neither the patient nor the dentist are aware of the condition and there is the potential of life threatening, emergency situations in what at first seem as simple, straightforward dental procedures. Therefore, the dentist must be able to recognize the clinical problem, differentiate between the different symptoms and initiate the proper management protocol. The most unstable endocrinal disorders that should be treated with great care are diabetes mellitus, mainly hypoglycemia, hyperthyroidism and adrenal insufficiency. The general practitioner dentist can treat patients suffering from these disorders providing the disease is well controlled and balanced and that the dental treatment is not very traumatic. PMID- 11852453 TI - [Cardiac arrest in dental offices. Report of six cases]. AB - The extreme medical emergency situation in the dental setting is cardiac arrest. The need to provide dental treatment to the medically compromised patients, suffering from very high risk heart diseases at special oral medicine hospital dentistry units, expose the dental and medical teams to the possibility of patients' death. Cardiac and cardiorespiratory arrest in these units faces the dentists with the need to perform basic and/or advanced cardiopulmonary resuscitation (CPR). Various etiologies are responsible for cardiac arrests. This article describes our experience and the outcome of six patients who have suffered cardiac arrests pre, during or post dental treatment in two special oral medicine centers. Two patients, suffering from severe congestive heart failure experienced fatal ventricular arrhythmia, both of them underwent CPR with early cardiac defibrillation, following which one patient completely recovered, and the other one expired. Two young and healthy patients experienced severe neurocardiogenic syncope with heart standstill for more than 40 seconds followed by spontaneous uneventful recovery. The fifth patient, who suffered from ventricular fibrillation as a result of an acute coronary ischemia, was resuscitated successfully. The last patient, a young woman, suffered from a severe status epilepticus causing bradycardia, which led to cardiac arrest, but recovered following CPR. All patients who did not recover spontaneously underwent methodical advanced CPR with early defibrillation. Only one patient out of the six died. PMID- 11852454 TI - [Legal and ethical considerations of emergencies in the dental office]. AB - The dentist has the ethical and legal responsibility to anticipate emergency situations in correlation with the patient's medical status. He has the obligation to do all in his power to prevent emergencies from happening and to be prepared to manage any emergency that might occur. This article also discusses the importance of monitoring and documentation. PMID- 11852455 TI - [Emergencies in dental medicine--introduction]. PMID- 11852456 TI - Productivity with word order and morphology: a comparative look at children with SLI and children with normal language abilities. AB - The study investigated the development of grammatical categories (verb and noun) in young language learners. Specifically, children's ability to mark correctly the semantic roles of agent and patient was investigated. Second, children's productive use of the present tense progressive -ing, past tense -ed, third person singular -s and plural -s inflection was investigated. Twenty-eight children with specific language impairment (SLI) with a mean language age of 35 months and 28 children with normal language (NL) with a mean language age of 34 months were exposed to four novel verbs and four novel nouns during ten experimental child-directed play sessions. The lexical items were modelled with four experimentally controlled argument structures. Furthermore, both groups of children were administered structured tests of grammatical morphology (involving both verbs and nouns). First, children in both groups did not correctly mark semantic roles for agent and patient, unless specific semantic roles had already been modelled in the linguistic input by the experimenter. Second, children's productive use of grammatical morphology in their spontaneous utterances was better for nouns than verbs. Importantly, however, children with NL were significantly better at marking verbs for past tense than in children with SLI, although both groups performed poorly overall. These findings are discussed in relation to current theories of normal and impaired language development, in particular the usefulness of tense marking as a clinical marker of preschool children with SLI. PMID- 11852457 TI - Development and disadvantage: implications for the early years and beyond. AB - Links have long been made between literacy and economic development, and recent governments in the UK have put great emphasis on the teaching of literacy to raise educational standards. There is substantial evidence to show that spoken and written language share some processes in common and that the development of literacy is supported by the development of spoken language. Anecdotal evidence from early years practitioners suggests that many children coming into early years education, particularly those from disadvantaged socioeconomic backgrounds, may not have the spoken language skills needed to develop reading and writing. This preliminary study aimed to investigate the extent to which the spoken language skills of children reared in poverty are depressed in comparison with the general population, and in comparison with their general cognitive abilities. Two tests were administered to children in their first term at nursery school to measure their spoken language skills and cognitive abilities: CELF-PUK and BAS II. More than half of the children were found to be language-delayed, although girls' receptive language abilities were significantly better than those of boys. Participants' language skills were also significantly depressed in comparison with their cognitive abilities. Government initiatives to raise awareness of spoken language in the early years are discussed, and implications for the future role of speech and language therapists working in the pre-school sector are considered. PMID- 11852458 TI - Psychological impact of the Lidcombe Program of early stuttering intervention. AB - The Lidcombe Program is an operant treatment for stuttering in preschool children for which favourable outcome and social validity data have been published. The treatment involves parental praise for stutter-free speech in children's everyday speaking environments, and occasional correction of stuttered speech. Theoretical perspectives on the origins of stuttering have prompted suggestions that the Lidcombe Program may have an adverse psychological impact on children. The present preliminary investigation sought to identify any evidence of such a systematic, pernicious trend, which might justify statistically powerful investigations of the issue with large subject numbers. Subjects were eight preschool children who were successfully treated with the Lidcombe Program. The Child Behavior Checklist (CBCL) detected any post-treatment behavioural markers of changes in children such as anxiety, aggression, withdrawal or depression. The Attachment Q-Set (AQS) measured any changes in the quality of the attachment relationship between child and mother over the course of treatment. These case studies revealed no evidence of a systematic trend in either. In fact, CBCL data suggested improvements in the children after treatment. It is concluded that there is no reason to doubt that the Lidcombe Program is a safe treatment. PMID- 11852459 TI - Using an interdisciplinary approach to training to develop the quality of communication with adults with profound learning disabilities by care staff. AB - The study examines the effects of training on care staff's communication to service users with profound and severe learning disabilities. An interdisciplinary therapeutic training programme aimed to improve care staff's cognitions about disability, to alter their communicative practice and to provide them with practical solutions to the communication difficulties of the service users. Participants were each video recorded with a single service user before and during training. The recordings formed the basis of the communication workshops. Both baseline and final recordings were evaluated using criteria based on verbal and non-verbal interactions. The criteria assessed alterations in the care staff's language use, verbal responses, and interpretation and praise of service users' communication. The use of posture, position, eye contact and gaze monitoring during interactions was also evaluated. The pre- and post-training analysis demonstrated alterations and improvements in care staff's use of verbal communication as well as gaze monitoring and position during interactions. A follow-up study 6 months later showed many of the post-training positive changes had been maintained or had continued to develop. It is suggested that the success and implementation of the communication training in the workplace was due to the collaborative therapeutic nature of the training package. The interdisciplinary and integrated training programme had facilitated a willingness to change and led to the development of a positive view of each other's practice. The recommendations from each of the therapist's training programme formed an integral part of the care staff's new care plans for each service user. This contributed to the continuing development the care staff's therapeutic role. PMID- 11852460 TI - Preferred communication modes: prelinguistic and linguistic communication in non speaking preschool children with cerebral palsy. AB - Seven non-speaking preschool children with severe cerebral palsy, 5-7 years of age, were studied with respect to the amount of prelinguistic versus linguistic modes of communication used in communicative interaction with a previously unknown adult. An attempt was also made to analyse this in relation to the childrens' physical, cognitive and linguistic capacities. No significant correlations were found, however. Given that all participants had reached preschool age and were of normal intelligence, the amount of linguistic communication used (12%) was much less than could be expected. Explanations for the results were suggested, including limited interaction possibilities with the environment due to motor dysfunction, which in turn might prevent development of linguistic skills necessary to take part in more complex communicative interaction later in life. PMID- 11852461 TI - A molecular genetic analysis of the Kidd blood group polymorphism in Taiwanese, Thais, and Filipinos. AB - In this study, we molecularly detect the Jka/Jkb polymorphism in Taiwanese, Thais, and Filipinos with PCR-restriction enzyme digestion. We designed the primers with mutagenic bases to create two restriction enzyme cutting sites (EcoN I and Xba I) simultaneously to confirm the genetic polymorphism of Jka/Jkb. One hundred and seven unrelated Taiwanese, 107 Thais foreign laborers, and 102 Filipino foreign laborers in Taiwan were analyzed. The frequency of Jka and Jkb is 0.51 and 0.49 in Taiwanese, 0.44 and 0.56 in Thais, and 0.45 and 0.55 in Filipinos, respectively. The molecular analysis of Jka/Jkb polymorphism is simple, accurate and could confirm the uncertain results obtained by serological typing. The method could be used for predicting fetal Kidd genotype from amniocyte DNA, and to avoid the hemolytic disease of newborns caused by Jk antibody. PMID- 11852462 TI - Recurrent hypertensive intracerebral hemorrhage among Taiwanese. AB - Hypertensive intracerebral hemorrhage (HICH) was once thought to be a one-time event with very rare recurrence, but recent studies have revealed that the recurrent hemorrhage due to hypertension is not unusual. The purpose of this study was to determine the clinical characteristics of HICH among Taiwanese. From June 1988 to December 1999, 1421 HICH patients were admitted to our hospital. Among them, 68 patients (4.8%) had recurrent HICH. We reviewed their medical records and computed tomographic findings. There were 46 males and 22 females (M/F = 2.1) with a mean age of 59.9 +/- 11.9 years at the onset of the second hemorrhage. The median interval between the first two hemorrhages was 22.5 months (range: 1-107 months). Most of the recurrence was within two years of the first hemorrhage (within 1 year in 27.9%, within 1-2 years in 25%). The location of the second hemorrhage was typical for HICH (putamen in 44.1%, thalamus in 33.8%, cerebellum in 5.9%, pons in 4.4%, and caudate nucleus in 1.5%) except for 7 patients (10.3%) who had lobar hematoma. Forty-nine patients (72%) had both hemorrhages located in the supratentorium and in most of them (40 patients) the recurrent HICH occurred contralaterally to the first one. Putaminal-thalamic pattern was the most common (23.5%), followed by the putaminal-putaminal pattern (20.6%). The mortality rate of the second HICH was 17.6%. Seven patients (10.3%) experienced more than 2 episodes of hemorrhages. This report found that the recurrent HICH was not rare among Taiwanese. A substantial proportion (19.1%) of recurrence was after 5 years. The male predominance and risk factor for recurrent HICH require further study. PMID- 11852463 TI - The effects on vesicourethral function following laparoscopic hysterectomy. AB - The aim of our study is to determine whether laparoscopic hysterectomy is associated with increased postoperative urinary symptoms and to assess the change in urodynamic parameters after operation. Forty-five women were arranged for laparoscopic hysterectomy (LH). Each patient received urinalysis, interview, and urodynamic study including uroflowmetry, filling and voiding cystometry and urethral pressure profilometry before and after hysterectomy. A total of 27 patients (60%) had urinary symptoms preoperatively. After operation, only 22 patients (48.9%) remained symptomatic. There was no significant change in the number of women with one or more voiding symptoms before and after surgery, but the incidence of urinary frequency and stress incontinence decreased significantly after laparoscopic hysterectomy (P < 0.05). In addition, maximal urethral closure pressure and maximal cystometric capacity showed significant increases after operation. They were 73.1 cm H2O (range: 49-114) vs 104.4 cm H2O (range: 60-147) (P < 0.001), and 363.3 ml (range: 287-423) vs 396.1 ml (range: 265-515) (P < 0.001), respectively. The result indicated that laparoscopic hysterectomy did not significantly increase the subjective or objective incidence of vesicourethral dysfunction. On the contrary, some patients might be cured of urinary frequency or stress incontinence postoperatively. PMID- 11852464 TI - Association of work status and mental well-being in new mothers. AB - The present study was to consider the relationship between work status and mental well-being of women at six weeks postpartum. One hundred and eighty-eight employed mothers and 82 unemployed mothers were recruited from the Kaohsiung city area in southern Taiwan to participate in this study. Five instruments were used to collect data: the demographic data form, Cohen's perceived stress scale, Cohen's interpersonal support evaluation list, Coopersmith's self-esteem inventory, and the Beck depression inventory. In results, unemployed mothers were younger, had fewer years of education, and lower socioeconomic status than employed mothers. To adjust for these differences in group characteristics, one way analysis of covariance (ANCOVA) was performed to determine the role of work on mental health factors. Unemployed mothers were found to be at significantly increased risk for depression in terms of prevalence and severity. They also showed significantly greater stress, reduced social support and lowered self esteem. That employment is mentally beneficial for post-partum women may be due to increased financial resources and provision of additional stress release outlets, social support resources and opportunities for affirmation of personal worth. Since this study is cross-sectional, some reasonable alternative causal explanations for these data were also discussed. PMID- 11852465 TI - Traumatic lumbar spinal subdural hematoma--a case report. AB - A 74-year-old woman suffered from lower legs weakness after a motor vehicle accident. Magnetic resonance imaging (MRI) on the seventh posttraumatic day, revealed a lumbar spinal subdural hematoma at the level of L4-5. After surgical intervention to remove the subdural hematoma, the patient made a complete recovery. PMID- 11852466 TI - Cystic left atrium myxoma--a rare case report. AB - Myxoma is the most common primary tumor of heart. The typical picture of myxoma under echocardiography is a solid, dense echo mass and left atrium is the most common site to find it. The cystic form of myxoma is vary rare. We report a patient who received echocardiographic examination under impression of mitral valve stenosis. A multilobulated cystic mass which was like a hydatid cyst was found in the left atrium and atrioventricular flow was affected by this mass. After tumor resection, myxoma with internal hemorrhage was proved by pathology. No further recurrent myxoma was found during follow-up echocardiographic examination. PMID- 11852468 TI - Giant tear retinal detachment after laser in situ keratomileusis--a case report. AB - A 42-year-old woman with a refractive error of -10.00S - 2.00C x 105 degrees in the right eye underwent laser in situ keratomileusis (LASIK) at our hospital. LASIK was performed using the Schwind excimer laser (Keratom Multiscan, Schwind, Kleinostheim, Germany) and Moria LSK--One manual microkeratome with a 130 microns ablation plate. The uncorrected visual acuity improved postoperatively, and the patient was very satisfied. However, nine months later, she complained of sudden loss of visual acuity in the right eye. Indirect ophthalmoscopy revealed a giant retinal tear extending from 10:00 to 2:00 o'clock position with retinal detachment and vitreous hemorrhage. We performed scleral buckling procedure (with silicone band encircling), vitrectomy, and fluid-gas exchange with air/SF6 mixture. The retina attached postoperatively and remained so during 3-month follow-up period, but cellophane maculopathy was noted. PMID- 11852467 TI - Pseudo-Meigs syndrome and elevated levels of tumor markers associated with benign ovarian tumors--two case reports. AB - Elevated tumor markers for a post-menopausal woman presenting with a multilocular adnexal mass, ascites, and pleural effusion were interpreted as being highly suspicious of malignancy. This paper describes two cases of ovarian tumors presenting with all signs of malignancy. Following surgical excision of the masses, and histopathological assay, a benign pure struma ovarii and a mucinous cystadenoma were diagnosed by pathologists. The immediate and complete resolution of symptoms were achieved post-operatively, and the previously-evident abnormal tumor markers rapidly declined to the normal range, the two tumors were subsequently classified as pseudo-Meigs' syndromes. PMID- 11852469 TI - [Human anthrax outbreak due to cow anthrax in Iwate Prefecture]. PMID- 11852470 TI - [A clinical study of tuberculous pleurisy]. AB - A clinical study of 38 patients (28 men and 10 women) with tuberculous pleurisy was conducted. The age of these patients ranged from 19 to 92 years, with an average age of 48.9 years. In 30 patients, the chief complaint was fever, and other common complaints included chest pain, dyspnea, and coughing. Bacillus tuberculosis was found in the pleural fluid of 7.9% of the patients. Tuberculous pleurisy was diagnosed histologically, based on pleural biopsy, in 23.7% of the patients. The diagnosis rate of pleural biopsy was 47.4%. There were no significant differences in results of blood and pleural fluid tests between idiopathic pleurisy and concomitant pleurisy, but the tuberculin skin test was positive in only 50% of the patients with concomitant pleurisy. The tendency was that the longer the time period between symptom onset and first examination, the greater the pleural fluid retention. The diagnosis rate of pleural biopsy was influenced by the severity of pleural fluid retention. A thoracic cavity drain was inserted for continuous drainage in 15 patients, and every patient underwent INH + RFP-based chemotherapy. Tuberculous pleurisy is an important disease among patients with pleural fluid retention, thus clinicians need to know how to treat this disease. PMID- 11852471 TI - [Retrospective analysis of Fusobacterium associated infections; experience at Aomori Prefectural Hospital with 108 cases from 1995 to 1999]. AB - We experienced 108 cases of Fusobacterium associated infections, including otolaryngeal, oral, pleuropulmonary, intraabdominal, skin and soft tissue infections, at Aomori Prefectural Hospital during The 5 year-period from 1995 to 1999. A total of 433 organisms, included 113 Fusobacterium spp. (80 Fusobacterium nucleatum, 18 Fusobaterium necrophorum, 5 Fusobacterium varium, 4 Fusobacterium mortiferum, 6 Fusobacterium spp.), were recovered with an average of 4.0 organisms per case of the 108 cases, 68% were mixed aerobic and anaerobic and yielded 185 anaerobic bacteria (2.5 per case) and 137 aerobic bacteria (1.9 per case) with an average of 4.4 per case. The remaining 32% were purely anaerobic and yielded 111 organisms with an average of 3.2 per case, Prevotella spp., Bacteroides fragilis group, Streptococcus milleri group, Enterobacteriaceae, Peptostreptococcus spp. Staphylococcus spp. were most frequently coisolated with Fusobacterium spp. PMID- 11852472 TI - [Hospital outbreak of viral gastroenteritis attributed to Norwalk-like viruses in Japan]. AB - BACKGROUND: Norwalk-like viruses (NLVs), RNA viruses in the family of Caliciviridae, are known as a pathogen of nonbacterial food-borne gastroenteritis associated with eating raw oysters. NLV can spread from person to person with strong infectivity and can cause large epidemics in communities, schools, nursing homes, and hospitals. Here, we describe an outbreak of gastroenteritis associated with NLVs, possibly introduced from outside the hospital, in four different wards on four different occasions between November 1999 and April 2000, in a university affiliated hospital in Tokyo. METHOD: Total 61 specimens (stool or vomitus) from 46 patients and staffs were collected. Reverse transcription-polymerase chain reaction (RT-PCR) test was performed on each specimen. Standard precautions were applied strictly as control. RESULT: NLV was detected in 40 specimens from 28 patients and staff (20 patients, 8 nurses). Attack rate of the patients were 0.19, nurses were 0.15, and doctors were 0.07. CONCLUSION: The spread of NLV from person to person is considered to be a major infection pathway in each ward. A nurse is at a high risk of infection as a patient in some wards. A stringent policy of control must be applied. PMID- 11852473 TI - [Studies on efficient methods for detection of Legionella species from environmental water by use of its selective media]. AB - To examine the optimal pH range for growth on media, growth of Legionella spp. on its selective media, BCYE alpha, WYO alpha and MWY agar media, in a pH range of 6.0-8.0 (at 0.5 intervals) was compared. The growth of two strains of L. pneumophila and one strain of L. micdadei on a WYO alpha agar supplemented with some selecting antimicrobial agents was markedly inhibited at all pH range except 6.0 and 7.0, suggesting a narrow optimal pH range for growth of these species compared to the BCYE alpha without selecting antimicrobioal agents. Vancomycin (VCM) added to the selective agar suppressed the growth of some Legionella spp. depending on the concentration. However, the extent of suppression was different among species and/or strains of Legionella spp. The selectivity for species other than Legionella spp. was also affected similarly by VCM concentration added to their media, suggesting that it is important to use proper amounts of the selecting antimicrobial agent depending on the species and/or strains of Legionella spp. or the other species in water samples. Amphothericin B (AMPH-B) added to a selective medium, MWY agar, in the concentration of 80 micrograms/ml hardly affected the growth of Legionella spp. examined, but effectively inhibited the growth of fungal strains identified as Aspergillus sp., Trichoderma sp., Scolecobasidium sp. and Mucor sp. which were isolated from cooling-tower water samples together with Legionella spp. Furthermore, the growth of a combination culture of one each of the 4 strains of isolated fungi and one each of the 3 strains of Legionella spp. was examined at various concentration of AMPH-B. Addition of AMPH-B to the selective medium at the concentration of 80 micrograms/ml suppressed the growth of spreading fungi, permitting the growth of Legionella spp. to allow efficient detection of the species. PMID- 11852474 TI - [Biological properties and drug susceptibility of verotoxin-producing Escherichia coli (VTEC) isolated from healthy goats in Okinawa Prefecture]. AB - Fecal samples from 116 healthy goats out of 25 randomly selected farms were examined for verotoxin-producing Escherichia coli (VTEC) during 1996 and 1998 in Okinawa Prefecture. VTECs were detected 204 (15.0%) from 1,361 E. coli strains, 36 (31.0%) goats out of 13 (52.0%) farms. Randomly selected 88 strains were further characterized according to VT types, serotypes, virulence markers, biochemical properties and drug susceptibility. VT types were classified as VT1 (46.6%), VT2 (6.8%), and VT1/VT2 (46.6%) by means of reversed latex agglutination test. The VTEC belonged to 18 different O serogroups: O1, O6, O22, O27, O48, O75, O76, O77, O78, O82, O91, O103, O111, O123, O125, O128, O146, and O158. Serotypes O91:H- (13 strains), O27:H- (10 strains), O22:H19 (6 strains) are considered to be predominant, whereas O serotypes O157 and O26 were not isolated. eaeA gene was detected only in 5 strains (5.7%):O103:H2 and O111:H-, in contrast, hlyA gene was found frequent in 45 strains (51.1%) belong to various O serogroups, except for O146 (8 strains). On the basis of 20 biochemical features in all isolates, characteristic patterns were divided into 14 distinct types:47 strains (57.3%) were classified as one type. The VTECs examined were resistant to streptomycin (26.7%), ampicillin (12.2%), kanamycin (8.9%), oxytetracyline (8.9%), and oxolinic acid (3.3%), respectively. The current results indicate that goats harbored VTEC at high frequencies and may be a potential reservoir of human VTEC infection. PMID- 11852475 TI - [Field trial of combined measles and rubella live attenuated vaccine]. AB - We investigated the antibody responses and clinical adverse reactions after immunization with live combined measles and rubella vaccine (HF vaccine) in 442 healthy children, aged 12-90 months of age. We obtained 368 paired sera. Among them, 363 were initially sero-negative against measles virus and 343 (94.5%) became sero-positive after immunization. Sero-conversion against rubella virus was demonstrated in 349 (96.7%) of 361 initially sero-negatives against rubella virus. We investigated the clinical adverse reactions in 406 recipients. In 102 (25.1%) recipients, febrile reaction (> 37.5 C) developed on the day 6.7 of vaccination on average, with a mean duration of 2.2 days. Only two (0.5%) developed high body temperature over 39.5 C. Skin rash was noted in 87 (21.4%) on day 7.1 of vaccination on average, with a mean duration of 4.8 days. Lymphoadenopathy was demonstrated in 12 (3.0%). Thus, measles and rubella combined vaccine was safe and sufficiently immunogenic as well as each monovalent one, having clinical advantage in immunization practice. PMID- 11852476 TI - [An autopsied case of septicemia due to Vibrio vulnificus]. AB - A 63-year-old male with liver cirrhosis due to type-C hepatitis virus was admitted on June 14, 1999 to our hospital with complaints of dyspnea, and blisters, swelling and purpuras on his legs. He had consumed raw fish one or two days before. He was already in a state of shock with sepsis and disseminated intravascular coagulation shortly after the admission. Although treatment with MEPM and MINO for sepsis, and daltepalin sodium, antithrombin III and gabexate mesilate for disseminated intravascular coagulation was begun within 12 hours, he died only 30 hours after admission. The causative organism was detected from the blood and the contents of blisters, and was determined as Vibrio vulnificus. On autopsy, Vibrio vulnificus was also detected from skin and muscular tissue of his legs, but necrotizing fasciitis were not apparently revealed. Coagulating necrosis and acute tubular necrosis were verified in intestine and kidneys respectively probably due to ischemic changes. Pseudolobuli were formed and a small hepatocellular carcinoma was detected in the liver. Vibrio vulnificus has two infection channels; one is oral intake and the other is an external wound. The former is said to become serious. It has a rather short period from the starting of the symptom to death, and is highly fatal. If this bacteria is suspected by the clinical coarse of the patients or the laboratory examinations, it is necessary to dose effective antibiotics in its early stage. And for prevention, susceptible patients must be informed of the existence of this disease and the necessity of adequately heating raw seafood. PMID- 11852477 TI - [An autopsy case of Ph1--positive acute lymphoblastic leukemia with disseminated infection of Fusarium solani]. AB - A 56-year-old woman with Ph1--Positive acute Lymphoblastic Leukemia was admitted to our hospital for induction chemotherapy in June 1999. The patient was presented with a central scotoma of left eye during treatment course and was given diagnosis of endophthalmitis. Thereafter she also developed skin induration and suffered from serious pneumonia. Amphotericin B administration was started because of high titer of beta-D-glucan, but soon discontinued due to its adverse effect. Blood cultures yielded colonies of fungus and it was identified Fusarium solani. Her general condition deteriorated with progression of pneumonia, and she died of respiratory insufficiency. Autopsy was performed, and its specimen revealed the disseminated infection of Fusarium solani (lung, eye, heart, kidney and skin). We should pay special attention to the fusariosis in Japan also. PMID- 11852478 TI - [Streptococcus salivarius meningitis after oral trauma by a skewer: a case report]. AB - Although Streptococcus salivarius is one of the normal flora in the oral cavity and gastrointestinal tract, the agent may cause bacteremia, meningitis, endocarditis and sinusitis under certain circumstances. We report a 3-year-old female with meningitis after oral trauma by a skewer due to penicillin resistant S. salivarius. The girl injured her throat accidentally with a skewer. Four hours later, she became febrile and came to our emergency room. Plain CT scan was normal, and cefalexin was prescribed. The next day, she had fever, lethargy, meningeal signs, and her cerebrospinal fluid (CSF) showed neutrophilic pleocytosis. The blood culture was negative, but the CSF culture was positive for S. salivarius. The minimal inhibitory concentrations (MIC) for panipenem, penicillin G, ampicillin, cefotaxime, ceftriaxone, vancomycin were 0.125 microgram/ml, 2 micrograms/ml, 2 micrograms/ml, 0.5 microgram/ml, 0.5 microgram/ml, 0.5 microgram/ml, respectively. Intravenous administration of panipenem betamiprom (PAPM/BP) 2 g/day for 7 days and 8 courses of dexamethasone 0.15 mg/kg/dose were effective, and she has had no apparent sequelae except for a slight abnormality in her electroencephalogram. Traumatic meningitis is often caused by S. pneumoniae, but may be also caused by the normal flora pathogens including S. salivarius. In addition, our case suggests that not only S. pneumoniae but also S. salivarius can be penicillin resistant. Taking the drug resistance into consideration, we have to be careful in choosing antibiotics for treating such patients. PMID- 11852479 TI - [Relationships between shedding of enterohemorrhagic Escherichia coli O157:H7 and feeds in cattle]. PMID- 11852480 TI - [Influenza vaccine: past, present and future]. AB - Japan has ever based its policy for controlling influenza on a strategy of vaccinating schoolchildren. Mass immunization program for schoolchildren began in 1962. The government discontinued the program in 1994, because of growing doubt about its effectiveness. However, it was recently reported that vaccinating schoolchildren against influenza provides protection and reduces mortality from influenza among elderly persons. Instead, Japanese government will begin a vaccination program for elderly persons, because the excess mortality rates increased, as the vaccination of schoolchildren was discontinued. Although vaccination rate of Japan is the lowest in developed countries at present, the supply of influenza vaccine increases double every year. PMID- 11852481 TI - Influenza in Canada: 2000-2001 season. PMID- 11852482 TI - Toxicology and carcinogenesis studies of fumonisin B1 (cas no. 116355-83-0) in F344/N rats and B6C3F1 mice (feed studies). AB - [formula: see text] Fumonisin B1 is a mycotoxin produced by the fungus Fusarium moniliforme, one of the major species found in corn. There are no known commercial or medical uses of fumonisin B1. Fumonisin B1 was nominated by the FDA Center for Food Safety and Applied Nutrition for study because of its occurrence in corn and corn-based products in the United States and its toxicity in field exposure of horses and pigs. Male and female F344/N Nctr BR rats and B6C3F1/Nctr BR (C57BL/6N x C3H/HeN MTV-) mice were exposed to fumonisin B1 (92% pure) in feed for 28 days or (greater than 96% pure) for 2 years. 28-DAY STUDY IN RATS: Groups of 10 male and 10 female rats were fed diets containing 0, 99, 163, 234, or 484 ppm fumonisin B1 for 28 days. There were no exposure-related deaths in rats. The mean body weights of the 484 ppm groups were significantly less (-16%) than those of the controls. Dietary concentrations of 99, 163, 234, and 484 ppm fumonisin B1 resulted in average daily doses of 12, 20, 28, and 56 mg fumonisin B1/kg body weight for males and females. Additional groups of male and female rats were exposed to the same concentrations of fumonisin B1 for 28 days for clinical pathology studies. The concentrations of creatinine, cholesterol, triglycerides, and total bile acids, as well as activities of the enzymes alanine aminotransferase, alkaline phosphatase, aspartate aminotransferase, and gamma glutamyltransferase, were generally significantly greater in the 484 ppm groups than in the control groups at all time points, indicating hyperlipidemia and a hepatic effect. Fumonisin B1 is an inhibitor of ceramide synthase, resulting in an interruption of de novo sphingolipid synthesis. This enzyme inhibition results in increased levels of sphinganine (or increased sphinganine:sphingosine ratio) in tissues and urine. Urinary sphinganine was increased in groups of males exposed to 163 ppm or greater, while urinary sphinganine was increased in all exposed groups of females. The kidney weights, relative to body weight, of all exposed groups of rats were less than those of the control groups, decreasing by approximately 11% in the females and 20% in the males. Apoptosis and degeneration of the kidney were observed in all exposed males and in most females exposed to 163 ppm or greater. The incidences of minimal to mild apoptosis, degeneration, and mitotic alteration of the liver were significantly increased in 234 and 484 ppm males and in females exposed to 163 ppm or greater. The incidences of bile duct hyperplasia were significantly increased in males and females in the 484 ppm groups. In the core study, male rats in all exposed groups and females exposed to 163 ppm or greater had significantly increased percentages of hepatocytes in one or more proliferative (non-G0) states. 28-DAY STUDY IN MICE: Groups of 12 male and 12 female mice were fed diets containing 0, 99, 163, 234, or 484 ppm fumonisin B1 for 28 days. There were no exposure-related deaths in mice. The mean body weights of the 484 ppm groups of males were significantly less than those of the controls. Feed consumption by males exposed to 484 ppm was less than that by the controls; dietary concentrations of 99, 163, 234, and 484 ppm fumonisin B1 resulted in average daily doses of approximately 19, 31, 44, and 93 mg/kg for males and 24, 41, 62, and 105 mg/kg for females. Additional groups of male and female mice were exposed to the same concentrations of fumonisin B1 for 28 days for clinical pathology studies. Cholesterol and total bile acid concentrations and alanine aminotransferase and alkaline phosphatase activities were increased at 484 ppm, indicating hyperlipidemia and a hepatic effect. Urinary sphinganine concentrations and sphinganine/sphingosine ratios were increased in 484 ppm male mice. In 484 ppm males and all exposed groups of females, the incidences of hepatocellular necrosis, diffuse periportal hypertrophy, and diffuse centrilobular hyperplasia, as well as hyperplasia of the bile canaliculi and Kupffer cells, were generally significantly greater than those in the controls. Core study males exposed to 99, 163, or 234 ppm had significantly increased incidences of hepatocellular cytoplasmic alteration. Hepatocytes of 484 ppm male mice and all exposed groups of female mice were induced into proliferative (non G0) states. 2-YEAR STUDY IN RATS: Groups of 48 male and 48 female rats (40 for 5 ppm groups) were fed diets containing 0, 5, 15, 50, or 150 ppm fumonisin B1 (males) or 0, 5, 15, 50, or 100 ppm fumonisin B1 (females) (equivalent to average daily doses of approximately 0.25, 0.76, 2.5, or 7.5 mg/kg to males and 0.31, 0.91, 3.0, or 6.1 mg/kg to females) for 105 weeks. Additional groups of four male and four female rats were exposed to the same concentrations as the core study animals and were evaluated at 6, 10, 14 or 26 weeks. Survival, Body Weights, and Feed Consumption Survival, mean body weights, and feed consumption of exposed male and female rats were generally similar to the controls throughout the study. Clinical Pathology Findings Sphinganine/sphingosine ratios were increased in the urine of 15, 50 and 150 ppm males and 50 and 100 ppm females exposed to fumonisin B1 for up to 26 weeks. The sphinganine/sphingosine ratios were also increased in kidney tissue of 50 and 150 ppm males (85- and 119-fold) and 50 and 100 ppm females (7.8- and 22-fold) at 2 years. Cell Proliferation Analyses Renal tubule epithelial cell proliferation was increased in 50 and 150 ppm male rats exposed to fumonisin B1 for up to 26 weeks. Renal tubule epithelial cell proliferation was marginally increased in 100 ppm females. Organ Weights and Pathology Findings Kidney weights of 50 and 150 ppm males were less than those of the controls at 6, 10, 14, and 26 weeks and at 2 years. Kidney weights of 100 ppm females were less than those of the controls at 26 weeks, and kidney weights of 15, 50, and 100 ppm females were less than those of the controls at 2 years. At 2 years, there was a significant increase in the incidences of renal tubule adenoma from none in the groups receiving 15 ppm or less to five of 48 in 150 ppm males. Renal tubule carcinomas were not present in male rats receiving 15 ppm or less and occurred in seven of 48 and 10 of 48 male rats in the 50 and 150 ppm groups, respectively. Incidences of apoptosis of the renal tubule epithelium were generally significantly increased in males exposed to 15 ppm or greater for up to 26 weeks. The incidences of focal renal tubule epithelial hyperplasia were significantly increased in 50 and 150 ppm males at 2 years. 2-YEAR STUDY IN MICE: Groups of 48 male and 48 female mice were fed diets containing 0, 5, 15, 80, or 150 ppm (males) or 0, 5, 15, 50, or 80 ppm (females) fumonisin B1 (equivalent to average daily doses of approximately 0.6, 1.7, 9.7, or 17.1 mg/kg to males or 0.7, 2.1, 7.1, or 12.4 mg/kg to females) for 105 weeks. Additional groups of four male and four female mice were exposed to the same concentrations as the core study animals and were evaluated at 3, 7, 9, or 24 weeks. Survival, Body Weights, and Feed Consumption Survival of males and females in the 15 ppm groups and of 5 ppm females was significantly greater and survival of 80 ppm males and females was significantly less than that of the control groups. Mean body weights and feed consumption of exposed mice were generally similar to the controls. Organ Weights and Pathology Findings Liver weights, relative to body weight, were increased 1.3 and 2.9-fold in 50 and 80 ppm females at 2 years. At 2 years, the incidences of hepatocellular adenoma in 50 and 80 ppm females were significantly greater than those in the controls and occurred with a positive trend. Similarly, the incidences of hepatocellular carcinoma increased from none in the groups receiving 0, 5, or 15 ppm fumonisin B1 to 10 of 47 females at 50 ppm and nine of 45 females at 80 ppm. The incidences of hepatocellular hypertrophy were significantly increased in 15, 80, and 150 ppm males and in 50 and 80 ppm females at 2 years. The incidences of hepatocellular apoptosis were significantly increased in 50 and 80 ppm females at 2 years. CONCLUSIONS: Under the conditions of these 2-year feed studies, there was clear evidence of carcinogenic activity of fumonisin B1 in male F344/N rats based on the increased incidences of renal tubule neoplasms. There was no evidence of carcinogenic activity of fumonisin B1 in female F344/N rats exposed to 5, 15, 50, or 100 ppm. There was no evidence of carcinogenic activity of fumonisin B1 in male B6C3F1 mice exposed to 5, 15, 80, or 150 ppm. There was clear evidence of carcinogenic activity of fumonisin B1 in female B6C3F1 mice based on the increased incidences of hepatocellular neoplasms. The sphinganine/sphingosine ratios were increased in the urine and the kidney tissue of rats receiving diets containing fumonisin B1. There was evidence of apoptosis and increased cell proliferation of the renal tubule epithelium in exposed rats, particularly in those groups of males that developed renal tubule neoplasms. Increased incidences of hyperplasia of the renal tubule epithelium also occurred in these groups of male rats. In mice exposed to the higher concentrations of fumonisin B1, males and females had increased incidences of hepatocellular hypertrophy and females had increased incidences of hepatocellular apoptosis. PMID- 11852483 TI - Lessons for critical care nurses on caring for the dying. PMID- 11852484 TI - Myocardial injury: contrasting infarction and contusion. PMID- 11852485 TI - Using clinical pathways in patients undergoing cardiac valve surgery. AB - Clinical pathways, protocols, and standing order sets help organize patients' care and eliminate variations created by practitioners' different preferences. Much attention is being focused on providing the most cost-effective care in the shortest time. Quality of care must be maintained during this process. Clinical pathways, protocols, and standing order sets help ensure that care is consistent and quality of care is maintained with the added benefit of a shorter stay in the hospital. PMID- 11852486 TI - Coronary artery bypass graft surgery without cardiopulmonary bypass: a review and nursing implications. PMID- 11852489 TI - Percutaneous sheath introducer systems used for venous access. PMID- 11852487 TI - Managing patients with acute thyrotoxicosis. PMID- 11852490 TI - The wonderful world of progressive care. PMID- 11852491 TI - Reflections of a neonatal intensive care nurse. PMID- 11852492 TI - The acute care nurse practitioner: an expanding opportunity for critical care nurses. PMID- 11852494 TI - Strategies for a successful return to school. PMID- 11852493 TI - Advancing sedation assessment to promote patient comfort. AB - As sedation assessment continues to evolve towards a more disciplined and standard part of clinical practice, the use of subjective sedation scales and objective sedation tools such as the BIS monitor continues to grow and show promise. The efforts of critical care nurses and their colleagues to better understand the value of these tools is integral to guiding their use and optimizing patient comfort. PMID- 11852495 TI - [Nutritional alterations in human immunodeficiency virus infection]. PMID- 11852496 TI - [Nutritional status in HIV infected patients]. AB - OBJECTIVES: Nutritional assessment in HIV infected patients must be a main objective in the treatment of this disease. The main objective of our work was to study the nutritional status with biochemical and anthropometrical parameters in a group of HIV infected patients. MATERIAL AND METHODS: A total of 119 patients were evaluated. In all patients the next parameters were assessed: age, sex, treatment with anti-retroviral drugs, performed an anthropometric evaluation (weight, height, tricipital skinfold, arm circumference, arm muscular circumference and body mass index) and biochemical evaluation (albumin, prealbumin, transferrin, total proteins, triglycerides, cholesterol, lymphocytes and count of CD4). RESULTS: Patients had an average age 37.9 +/- 9.9 years, weight 64.5 +/- 13.2 kg and body mass index 22.5 +/- 3.5. The values of total proteins, albumin, prealbumin y transferrin were all normal, only patients with AIDS had low levels of transferrin (262.5 +/- 49.2 mg/dl vs 277.8 +/- 87.5; p < 0.05). Percentil distribution of anthropometric parameters showed a deep depletion in muscular protein compartment, 53.1% of patients had skinfold under P 50, 91.8% had arm circumference under P 50, and 91.8% arm muscular circumference under P 50. High levels of triglycerides were detected in 30.5%, these levels are related with the HIV phase and the number of antiretroviral drugs. CONCLUSION: Nutritional status in HIV infected patients is good in our population, only a deep depletion in muscular proteic compartment was detected. Transferrin was decreased in AIDS patients. High levels of triglycerides were secondary to the number of antiretroviral drug taking by patients. PMID- 11852497 TI - [Correlation between CD4 lymphocytes and viral load in patients with HIV and tuberculosis]. AB - OBJECTIVE: A retrospective study of the correlation between CD4 lymphocytes and the viral load in 16 HIV-patients with tuberculosis. MATERIAL AND METHODS: The clinical forms of presentation of tuberculosis were classified according to the location/s of the disease into: pulmonary, distinguishing between typical pulmonary and atypical according to the radiological pattern; extrapulmonary; mixed forms: pulmonary and extrapulmonary; and miliary tuberculosis. RESULTS: Tuberculosis was exclusively pulmonary in 7 cases (44%), all were atypical; extrapulmonary 6 (38%); mixed 2 (12%); and miliary 1 (6%). The mean CD4 lymphocyte count was 111.1 (range 5-360), in 11 (69%) the counts were below 200 cells/mm3. The lowest CD4 count was in the mixed forms with a mean of 45 cells/mm3 whilst the highest was obtained in pulmonary forms with a mean of 128.3 cells/mm3. The mean viral load was 4.82 log (range, 0-5.93), the highest load was for mixed forms with a mean 5.69 log, whereas the lowest load was for pulmonary forms with a mean of 4.19 log. No significant correlation was observed between CD4 lymphocytes and viral load (correlation coefficient--0.1163). CONCLUSIONS: Though no significant correlation was observed, a high CD4 was associated to a low viral load and inversely a low CD4 with a high viral load. PMID- 11852498 TI - [Clinical characteristics of patients with non-hodgkin lymphoma in Arnau de Vilanova Hospital, Valencia, Spain]. AB - OBJECTIVE: To define the clinical characteristics of patients with non-Hodgkin lymphoma (NHL) in general and by histological subtypes based on the REAL classification. PATIENTS AND METHODS: From a total of 210 case histories of patients with NHL, 188 presenting a complete pathology report were selected. The latter was either based on the REAL classification or was converted to it by an expert. The clinical features of the patients at the time of diagnosis were also recorded. RESULTS: Significant differences were observed in the distribution by sex among mantle cell lymphomas, with a predominance among males (p = 0.005). The youngest patients were diagnosed of Burkitt lymphoma and B lymphoblastic lymphoma. Patient status improved significantly (p = 0.0001), with a predominance of ECOG 0, and the stage of the patients studied was 74.1% (stage III/IV) versus 25.9% (stage I/II). CONCLUSIONS: The different characteristics of patients with NHL suggest the need for a single classification with diagnosis by histological subtype. PMID- 11852499 TI - [Auto-induced eosinophilic panniculitis: a diagnostic dilemma]. AB - Eosinophilic panniculitis is characterized by a prominent infiltration of subcutaneous fat with eosinophils. The clinical spectrum of lesions that display pathologic changes of eosinophilic panniculitis is diverse, but nodular lesions are the most common. The factitial eosinophilic panniculitis are exceptional, show a pleomorphic pattern of findings and are associated with personality disorders. Its main challenge is to establish the correct diagnostic. We report an unusual case of factitial nodular febrile panniculitis of the left forearm and proximal arm in an 16 year-old girl. She admitted that the lesions were self inflicted by autoinjection of several farming products as attempted suicide. We describe the clinical, radiological and histopathological features as well as the differential diagnosis of this type of panniculitis. PMID- 11852500 TI - [Idiopathic mediastinal fibrosis associated with hypercoagulability: a case report]. AB - Mediastinal fibrosis is a rare and unknown disease characterized by the presence of a fibrotic mass in the anterior mediastinum that can invade close structures as gullet, trachea, recurrent nerve, superior cava vein etc. Only 1-2% of patients with mediastinal fibrosis associate superior cava vein obstruction. When it appears, it is produced by extrinsic compression for fibrotic magma in almost all the cases. A case of hypercoagulability disorder associated with mediastinal fibrosis and superior cava vein obstruction by thrombosis (intrinsec mechanism), and the satisfactory evolution after the treatment with oral anticoagulation, corticosteroids and tamoxifen is presented. PMID- 11852501 TI - [Lung sarcoidosis following instillation of mitomycin C in the urinary bladder]. AB - Intravesical prophylaxis against recurrence of urinary bladder carcinomas using mitomycin C (MMC) has proved to be and effective treatment with few side effects. Previously only two cases of lung toxicity after instillation of MMC into the urinary bladder has been described. We report a 65-year-old man in whom lung sarcoidosis occurred after intravesical administration of MMC. This association has not been reported to date. The clinical picture and the pathogenesis of this lung disease are discussed. PMID- 11852502 TI - [Usefulness of skin tests (tuberculin and delayed-type hypersensitivity reaction) in patients with HIV infection]. AB - The aim of this study is to asses the management and application of skin tests (tuberculin and delayed-type hypersensitivity reaction) in patients with HIV infection. PMID- 11852503 TI - [Bioethical principles: are they applied in terminal illness situations?]. AB - Comprehensive practice of biomedical ethics principles (beneficence, non maleficence, autonomy and justice) would guarantee appropriate assistance to patients with terminal diseases. Unfortunately today, these principles aren't put into practice in a systematic way. In order to guarantee quality care to this particular group of patients, it is necessary that all health care professionals dealing with terminal illness receive a comprehensive education in bioethics matters and that an appropriate methodology is followed in order to resolve subsequent conflicts. Furthermore, public institutions should be involved so that Palliative Care Units provide assistance to the entire population rather than select cases. If we can't provide comprehensive care, we are working against the basic bioethics principles, especially the fundamental principle of justice. PMID- 11852504 TI - [Respiratory infection caused by Sphingobacterium multivorum]. PMID- 11852505 TI - [Gastric metastasis from lung carcinoma]. PMID- 11852506 TI - [Myelodysplasia with chromosome 7 deletion associated to tuberculosis and hepatitis C: a pattern of immunodepression synergism?]. PMID- 11852507 TI - [Pseudotumoral hepatomegaly as a presentation of amyloidosis]. PMID- 11852508 TI - [Cutaneous Paget's disease and adenocarcinoma]. PMID- 11852509 TI - [Kingella kingae endocarditis]. PMID- 11852510 TI - [Disseminated infection due to Streptococcus constellatus in an HIV patient with negative viral load]. PMID- 11852511 TI - [Asymptomatic splenomegaly from splenic hemangioma]. PMID- 11852512 TI - [Response of thyrotropinoma to somatostatin analogues: report of a case]. PMID- 11852513 TI - [Fournier gangrene and other necrotizing infections]. PMID- 11852514 TI - [Is free PSA useful for more than publishing articles?]. PMID- 11852515 TI - [Surgical treatment of hydrocele with local anesthesia: 10-year experience]. AB - OBJECTIVE: To review our experience with surgical management of hydrocele under local anesthesia. METHODS: From January 1991 to March 2001, 76 patients with hydrocele (6 bilateral) were treated in our department. Patients were monitored in the OR (EKG, BP and pulseoxymetry); a peripheral line was inserted. Twenty minutes before the procedure, midazolam (5 mg), meperidine (50 mg) and atropine (0.5 mg) were administered i.m. for sedation. The spermatic cord and the area of the scrotal wall to be incised were injected with 2% mepivacaine (10-20 cc). Dissection and excision of the vaginalis sac was performed. RESULTS: Anesthetic tolerance was very good in 71 patients (93%) and unsatisfactory in 5 (7%). The following complications were observed: scrotal hematoma (4 cases; 5%), infection (3 cases; 4%), bradycardia and hypotension (2 cases; 3%). Only 5 patients (7%) required hospitalization for more than 24 hours. CONCLUSIONS: Surgical management of hydrocele can be performed under local anesthesia, thereby avoiding the morbidity of more aggressive anesthetic techniques. This treatment procedure should be considered in the smaller departments with limited resources. PMID- 11852516 TI - [Primary neuroectodermal tumor (PNET) of the kidney: 26 cases. Current status of its diagnosis and treatment]. AB - OBJECTIVE: To report a case of primitive neuroectodermal tumor (PNET) of the kidney and review the literature and the 25 cases previously reported as PNET. METHODS: A 39-year-old man who consulted for nephric colic is described. Ultrasound evaluation disclosed a mass arising from the left kidney. The clinical, radiological and pathologic features, treatment and differential diagnosis of small cell tumors are discussed, as well as the important role of immunohistochemical techniques (positive staining with O13 or 12E7 antibodies) and cytogenetic analysis [a characteristic chromosomal translocation t(11;22) (q24;q12) or variant translocation, such as t(21;22) (q22;q12), may be detected by fluorescence in situ hybridization (FISH) or polymerase chain reaction-reverse transcriptase (PCR-RT)]. RESULTS: Survival of our patient was 20 months. Only three of the 25 previously reported cases had a longer survival: 60, 48 and 24 months. Mean survival was 10 months. 95.24% of the cases were positive for NSE. Immunostaining (CD99) was performed in 16 patients and was found to be positive in all cases. Cytogenetic and molecular analyses were performed in 11 cases; PCR RT was negative in two, as well as in the case described herein. CONCLUSIONS: PNET is a highly aggressive neoplasm that tends to recur locally and to metastasize. Despite the poor response to standard therapy combining surgical resection, postoperative irradiation and chemotherapy, the results might change due to current research on genetic therapy based on creating antisense oligonucleotides against the EWS-FLI 1 fusion gene. PMID- 11852517 TI - [Synchronous tumor of the upper urinary tract with bladder cancer. Opportunity for nephroureterectomy and block-cystectomy]. AB - OBJECTIVE: One of the basic characteristics of urothelial carcinoma is its tendency to synchronous or metachronous multifocality. Thus the need to explore the entire urinary tract of patients with urothelial neoformations. The aim of this article is to study the tumors of the upper urinary tract that appear synchronously with infiltrating carcinoma of the bladder. The clinicopathological characteristics and the morbidity and mortality of en bloc surgery of both tumors are analyzed. METHODS: A retrospective study was carried out on 170 radical cystectomies for infiltrating bladder tumor performed in our department over a 13 year period. Patient history, clinicopathological characteristics, complementary tests, type of surgery performed, postoperative complications and follow-up were analyzed. RESULTS: Tumor of the upper urinary tract appeared in 14 (1 bilateral) of these patients and were synchronous in 10 cases. All patients were male; mean age 63 years. Three were localized in the pelvis, 2 in the proximal ureter and 6 in the distal third. Diagnosis was made by IVP in 6 patients and by US and antegrade pyelography in the other 4 patients. Nephroureterectomy and radical cystectomy were performed en bloc in 8 cases; 6 had a Bricker procedure and 2 ileal substitution. Salvage radical cystectomy + distal ureterectomy were performed in the other two patients. Two patients submitted to en bloc surgery had postoperative complications; one presented prolonged ileua and the other required surgery for retroperitoneal hemorrhage. The two patients submitted to palliative surgery died of and sepsis during the postoperative period. At 33 months' mean follow-up, 3 patients have shown tumor progression. CONCLUSIONS: There is a high proportion of synchronous tumor of the upper urinary tract in our series of patients with infiltrating carcinoma of the bladder undergoing radical cystectomy, therefore we consider it necessary to explore the entire urinary system. Surgical removal of both tumors en bloc does not increase the morbidity and mortality. PMID- 11852518 TI - [Prevalence of prostatic intraepithelial neoplasia in Spain]. AB - OBJECTIVE: To study the prevalence of high grade prostatic intraepithelial neoplasia in a Spanish population and to compare it with the prevalence in Caucasians of other countries and Afro-Americans. METHODS: 162 prostates obtained at autopsy from Spanish men aged 20-80 years, were fixed in 10% formalin and slices perpendicular to the posterior margin were made every 3-4 mm along its entire length. All blocks were embedded in paraffin and examined microscopically. Mapping of focality and site of the high grade prostatic intraepithelial neoplasia was done for each case. The Wayne University autopsy study was used for comparison of the prevalence in other countries and races. RESULTS: 146 prostates from men with a mean age of 48.5 years were considered valid for histological analysis. There were 42 high grade prostatic intraepithelial neoplasia; 20 were focal and 22 multifocal. By age group, the prevalence of high grade prostatic intraepithelial neoplasia were 7.1%, 14.7%, 28.5%, 33.3%, 45.4% and 51.8% for the 3rd, 4th, 5th, 6th, 7th, and 8th decades. CONCLUSIONS: High grade prostatic intraepithelial neoplasia begins to manifest in the Spanish population after the 3rd decade. It is usually focal and peripheral, and significantly increases with age and becomes multifocal. Its prevalence in the Spanish population is moderately lower than in American Caucasians and significantly lower than in Afro Americans. PMID- 11852519 TI - [Current role of percutaneous renal surgery at a service of general urology]. AB - OBJECTIVE: To evaluate the utility and need for percutaneous surgery in the General Urology Department according to its applicability and efficacy. METHODS: 40 percutaneous surgery procedures were performed over a period of 11 years (average 3.6 procedures/year), which accounts for 0.60% of all surgical procedures performed. These were carried out for pyelic stone (10), caliceal stone (12), pyelic stone in horseshoe kidney (2), pyelic stone + UPJ stricture (4), UPJ stricture (7), UPJ re-stenosis post-pyeloplasty (1), cystectomy (3), diagnostic percutaneous surgery (1). RESULTS: Good results were achieved in 89.2% of the cases with lithiasis and there were 3 failed attempts (10.7%). Cystectomy achieved a 100% success rate; residual cavity persists in all the cases but there has been no recurrence at 5 years. Endopyelotomy was successful in 41.6% of the cases and stricture recurred in 58.3%. Overall, there were 4 complications without major consequences, although the procedure could not be completed in two cases. CONCLUSIONS: Although there is little opportunity to apply the technique, it is currently necessary because it achieves unquestionably good results in specific cases. PMID- 11852520 TI - [Urinary lithiasis secondary to indinavir in an HIV-positive patient]. AB - OBJECTIVE: Indinavir is a protease inhibitor used in the treatment of HIV with a lithogenic capacity as a urological side effect. The pathogenesis, diagnosis and treatment of indinavir urolithiasis are briefly reviewed. METHODS: A 37-year-old male, seropositive for HIV on treatment with indinavir, lamiduvine and zidovudine, consulted for colicky left lumbar pain, nausea, vomiting and dark urine for the past three days. RESULTS: Patient evaluation showed a nonfunctioning left kidney and ureterohydronephrosis of unknown origin. URS showed a yellowish, friable material with a mucinous appearance that occupied the entire lumen of the ureter. Fragmentation was achieved with the lithotriptor probe. Six months later the patient had fully recovered and was asymptomatic. CONCLUSIONS: The incidence of protease inhibitor-induced urolithiasis is increasing. This condition should be distinguished from uric acid calculi whose treatment will aggravate the indinavir urolithiasis. PMID- 11852521 TI - [Giant chondroid syringoma of the scrotum. First reported case]. AB - OBJECTIVE: Various tumors have been described in the scrotal area arising from skin and the underlying tissues: nevus, dermoid and epidermoid cysts, epidermoid carcinomas, lipomas, leiomyomas, angiokeratomas, lymphangiomas, granular cell tumors, granuloma, malignant tumors of the peripheral nerve tissue, and some 'pseudotumors' such as fibromatosis and nodular calcinosis. We describe for the first time a sweat gland tumor, which is also remarkable for its unusually large size. METHODS/RESULTS: A 76-year-old patient consulted for a painful left scrotal tumor that he had noted for some time and that had slowly and gradually grown. Ultrasound assessment of the GU system confirmed the presence of a 4.2 cm left, solid paratesticular mass that was removed under local anesthesia. CONCLUSIONS: Chondroid syringoma is a tumor arising from the sweat gland that is usually localized to the head and neck. It has an excellent prognosis and recurrence has only been described in patients in whom the tumor had not been completely resected. PMID- 11852522 TI - [Low-grade oncocytic adrenal carcinoma]. AB - OBJECTIVE: To report a case of left adrenocortical neoplasm. METHODS: A case of left adrenocortical neoplasm measuring 16 x 10 x 7 cm in a 47-year-old male is presented. After puncture biopsy, a left adrenalectomy through the abdominal approach was performed. RESULTS: Immunohistochemical studies showed reactivity to the anti-Melan-A A103 antibody, suggesting an adrenal origin of the tumor cells. Strong reactivity to the mES-13 mitochondrial antibody, which is characteristic of oncocytes, made ultrastructural studies unnecessary. CONCLUSIONS: In this type of tumor, an increased mitotic activity and necrosis are suggestive of malignancy. Despite the foregoing, most of this type of lesions are clinically benign and nonfunctioning. PMID- 11852523 TI - [Uretero-colonic fistula in non-functioning ureter]. AB - OBJECTIVE: To present a case of ureterocolonic fistula secondary to acute sigmoid diverticulitis in a nonfunctioning ureter due to a previous nephrectomy. METHODS/RESULTS: A 68-year-old patient that had undergone nephrectomy due to xanthogranulomatous pyelonephritis two years earlier, consulted for long-standing non-specific abdominal pain. Radiological evaluation showed a pneumogram pattern in the ureteral stump associated to a pelvic mass. The patient underwent surgery for a suspected uretero-intestinal fistula. The intraoperative findings and anatomopathological study demonstrated a uretero-sigmoid fistula due to diverticular disease of colon. CONCLUSIONS: Uretero-intestinal fistulas present unimportant clinical features. Radiological assessment and a clinical suspicion are important to diagnosis. PMID- 11852524 TI - [Sclerosing Sertoli cell tumor of the testis in an HIV patient]. AB - OBJECTIVE: To describe an additional case of an uncommon variant of Sertoli cell testicular tumor. METHODS/RESULTS: A 34-year-old male infected with the human immunodeficiency virus presented with a right testicular tumor he had noted one year earlier. A right inguinal orchidectomy was performed. Histological examination demonstrated sclerosing Sertoli cell tumor. CONCLUSIONS: Sertoli cell tumor is a rare variant of testicular tumor from sexual cords. Although it is uncommon, histological variants have been described: classical Sertoli cell tumor, large calcifying cells and the sclerosing variant. The case of sclerosing Sertoli cell tumor described herein is a variant of which 11 cases have been reported in the world literature. The fact that our patient is HIV-positive makes this case even rarer. PMID- 11852525 TI - [Splenosis: underdiagnosed entity]. AB - OBJECTIVE: To report a case of renal splenosis in a patient that had previously undergone splenectomy due to traumatic rupture. METHODS/RESULTS: A 42-year-old patient consulted for pancreatitis. A CT scan disclosed a mass behind the tail of pancreas, MRI assessment with gadolinium enhancement showed a left renal mass which was surgically resected. Histopathological analysis of the surgical specimen demonstrated splenic tissue. CONCLUSIONS: Splenosis should be considered in splenectomized patients with lesions compatible with tumors and should be distinguished from hyperplasia of accessory spleens. PMID- 11852526 TI - [Paratesticular recurrence of renal carcinoma]. AB - OBJECTIVE: To describe the clinical and histological findings of a case of renal cell carcinoma metastatic to the testes, an uncommon site of metastasis of this tumor type that can cause difficulty in making the differential diagnosis and consequently, in the staging and treatment of the disease. METHODS/RESULTS: A 65 year-old patient diagnosed of renal cell carcinoma presented a paratesticular mass. Histopathological examination of the orchidectomy specimen showed proliferation of clear cells arranged in a diffuse pattern, with intimate intertwining with vascular structures. Tumor cells were strongly positive for vimentin, CAM5.2 and EMA. CONCLUSIONS: Although renal cell carcinoma rarely metastasizes to the testes, it should be considered in the differential diagnosis of testicular masses. Histological examination is essential to diagnosis and correct management. PMID- 11852527 TI - [MALT lymphoma of the bladder. Report of a case]. AB - OBJECTIVE: To describe and discuss primary malignant lymphoreticular proliferative tumors of the bladder. METHODS: A case of a 70-year-old woman with hematuria, dysuria, malaise and 5-kg weight loss is presented. RESULTS: Pelvic ultrasound examination showed an 8 x 7 x 8 cm solid and cystic mass adjacent to the bladder and uterus. Cystoscopic biopsy disclosed a low grade B cell non Hodgkin lymphoma. Disappearance of the mass was achieved with 6 cycles of chemotherapy and radiotherapy. Four years thereafter the patient remains free of disease. CONCLUSIONS: Primary malignant lymphoma of the bladder is uncommon. The anatomopathological study is essential to differential diagnosis from other diseases. The response to chemotherapy and radiotherapy are excellent and permits bladder preservation. PMID- 11852528 TI - [How much tissue is necessary to diagnose mucinous adenoma of the prostate?]. PMID- 11852529 TI - [Urogenital malacoplakia]. PMID- 11852530 TI - Intravesical epirubicin treatment following TUR in superficial bladder tumours. AB - OBJECTIVE: To evaluate the effectiveness and tolerability of intravesical epirubicin treatment to prevent recurrence and progression of superficial bladder tumor after TUR. METHODS: 44 patients with superficial bladder tumor, 28 primary and 16 recurrent cases were given 50 mg intravesical epirubicin once a week for 8 weeks after TUR. Recurrent cases had not been treated by any intravesical agent before. Follow-up was done by cystoscopy, urine cytology and random biopsies. RESULTS: Local recurrence after treatment was found in 14.28% and 18.75%, and recurrence in another area was found in 21.42% and 31.25% of the primary and recurrent tumor groups respectively. Total recurrence in both groups was 40.91%. No systemic toxicity was observed and local side effects were treated symptomatically. CONCLUSION: Intravesical epirubicin after TUR for superficial bladder tumors especially for the primary ones, is a reliable method that provides effective prophylaxis with tolerable side effects. PMID- 11852531 TI - Novel steroidal pyrimidyl inhibitors of P450 17 (17 alpha-hydroxylase/C17-20 lyase). PMID- 11852532 TI - Introduction of a methyl group in alpha- or beta-position of 1-heteroarylethyl-4 phenylpiperazines affects their dopaminergic/serotonergic properties. AB - 1-(2-Heteroarylalkyl)-4-phenylpiperazines containing methyl group in either the alpha- or the beta-position of the side alkyl chain were synthesized as racemic mixtures. They were evaluated for in vitro binding affinity at the D1 and D2 dopamine and 5-HT1A serotonin receptors using synaptosomal membranes of the bovine caudate nucleus and hippocampus, respectively, as a source of the corresponding receptors. Tritiated SCH 23390 (D1 receptor-selective), spiperone (D2 receptor-selective), and 8-OH-DPAT (5-HT1A receptor-selective) were employed as the radioligands. None of the new compounds expressed significant affinity for the D1 receptor. Introduction of the methyl group into the beta-position of the parent molecules increased the affinity for the D2 receptor (10b-13b), and decreased the affinity for the 5-HT1A receptor with the exception of imidazole (11b) which was a rather efficient displacer of 8-OH-DPAT. Most potent of the newly synthesized compounds in [3H]spiperone assay were compounds (+/-)6-[1 methyl-2- (4-phenylpiperazin-1-yl)-ethyl]-1,4-dihydroquinoxaline-2,3-dione (10b), Kd = 6.0 nM and (+/-)5-[1-methyl-2-(4-phenylpiperazin-1-yl)-ethyl]-1,3 dihydrobenzoimidazol- 2-thione (13b), Kd = 5.3 nM. However, compounds containing methyl group in alpha-position (10a-13a) of the parent molecules expressed a decreased affinity for the D2 receptor, while the affinity for the 5-HT1A receptor remained in the same range of concentrations as that of closely related achiral parent compounds (14-17) run in the same binding assays as references. PMID- 11852533 TI - Synthesis, antimicrobial, and alkylating properties of 3-phosphonic derivatives of chromone. AB - Dimethyl 2,6-dimethyl-4-oxo-4H-chromen-3-yl-phosphonate (1a) and dimethyl 6 methyl-2-phenyl-4-oxo-4H-chromen-3-yl-phosphonate (1b) were synthesized and reacted with primary aliphatic amines to yield title compounds 4-6. Their antibacterial properties against Gram-positive and Gram-negative bacteria strains were tested by the MIC method. Four of seventeen tested compounds (1d, 3, 4a, and 4b) exhibit detectable activity against S. aureus. Some representative examples of newly synthesized compounds were tested for their alkylating properties in vitro in the Preussmann test. Compounds 1a, 1c, 1d, 3, 5d, and 6a possess highly alkylating activity toward standard derivative 4-(4'-nitrobenzyl)pyridine (NBP). PMID- 11852534 TI - Effect of oxovanadium(IV) complexes on nondiabetic and streptozotocin-diabetic rats. AB - The effects of vanadium complexes with organic ligands, [VO(phen)2]SO4.3H2O, [VO(bpy)2]SO4.2H2O, and [VOCl2(Hmcp)2H2O], on blood glucose and plasma lipid levels were studied in nondiabetic and streptozotocin-diabetic rats and compared to that of [VO(mal)2] (the reference compound). The present results provide evidence that the compounds examined possess lower toxicity than [VO(mal)2]. One of the compounds examined, viz. [VO(bpy)2]SO4.2H2O, decreases, statistically significantly, the glucose level and a second one, viz. [VOCl2(Hmcp)2H2O], decreases, also significantly, the total cholesterol level. PMID- 11852535 TI - Synthesis and analgesic-antiinflammatory activities of novel acylarylhydrazones with a 5-phenyl-4-R-3-pyrrolyl-acyl moiety. AB - A new series of arylidene 5-phenyl-4-R-pyrrole-3-carbohydrazides 1a-j were prepared and evaluated for their analgesic-antiinflammatory activities. All synthesized compounds showed a significant analgesic action in mice after intraperitoneal administration at a dose of 100 microM/kg. Two of these, 1b, (4' methylbenzylidene)-5-phenyl-1H-pyrrole-3-carbohydrazide, and 1d, (4' chlorobenzylidene)-5-phenyl-1H-pyrrole-3-carbohydrazide, were found to be more potent as antinociceptive agents respect to dipyrone and indometacin, used as reference drugs. Among compounds 1, only 1b showed a moderate antiinflammatory effect in rats while 1d proved to be a potent non antiinflammatory analgesic. PMID- 11852537 TI - [Survey on characteristics and diffusion of nosocomial infection control committees at Italian public hospitals]. PMID- 11852536 TI - Synthesis of vinca alkaloids and related compounds, Part XCVI. Nitration study of vinblastine-type bisindole alkaloids. AB - The bisindole alkaloids vinblastine, vincristine, and N-formyl-leurosine were nitrated and subsequently converted to amino derivatives. In the case of compounds 5e and 5b cytotoxic activity has been found for non-small cell lung cancer and breast cancer in the concentration range tested (10(-5)-10(-9) M). 5b also showed potency in the screen for colon cancer and leukemia. PMID- 11852539 TI - [Producer control and microbiological risk. Commercial and service catering in an area of high tourism impact (Elba Isle - Piombino]. PMID- 11852538 TI - [Epidemiologic and virologic surveillance of flu in Puglia]. PMID- 11852540 TI - [Type 1 diabetes in Lazio. Note 2 -- case control study of risk factors]. PMID- 11852541 TI - [Evaluation of the accuracy of hospital discharge records for the surveillance of bacteremia]. PMID- 11852542 TI - [Ageing of the work force and the opportunity of working after retirement]. PMID- 11852543 TI - [Work-related injuries among minors. Italian multicenter study, 1994-1998]. PMID- 11852544 TI - HPV-direct in situ PCR: an advanced molecular tool in the screening of cervical cancer. PMID- 11852545 TI - Perspectives. Proposal is down payment on long-term Bush Medicare vision. PMID- 11852546 TI - Telemedicine: is there a market somewhere out there? PMID- 11852547 TI - [Screening for diabetic maculopathy]. AB - Diabetic maculopathy (DMP) is an important but curable of eyesight losses in diabetic with type 1 and 2 diabetic mellitus (DM). In the submitted work the author investigates the pathogenesis of diabetic macular oedema (DME). Classification, diagnosis and standard of treatment of DMP, incl. the different therapeutic procedure used in concurrent DMP and proliferative diabetic retinopathy in DM type 1 and 2. PMID- 11852548 TI - [Treatment of severe caustic injuries of the anterior segment of the eye with limbal stem cell transplantation, penetrating keratoplasty and amnionic membrane transplantation]. AB - The concept of limbal stem cells (LSC) opened new possibilities for the treatment of severe caustic damage of the anterior segment. These possibilities include transplantation of LSC, perforating keratoplasty, possibly transplantation of the amniotic membrane. In three patients with severe caustic damage of the anterior segment of the eye the authors implemented transplantation of LSC, perforating keratoplasty and in one patient also transplantation of the amniotic membrane. Where only one eye was damaged LSC for transplantation were obtained from the sound eye (autotransplantation, in case of damage of both eyes LSC were obtained in one case from a close relative in the second case from a cadaverous eye from a corneal bank (Allotransplantation). In the patient with autotransplantation of LSC, perforating keratoplasty and transplantation of the amniotic membrane the result was very good. The corneal disc remained clear 1.5 years after operation, the conjunctival sac was free from synbleropharons and vision improved from movement of the hand to 0.5. In the two patients with allotransplantation of LCS and perforating keratoplasty, LCS did not become incorporated despite immunosuppressive treatment and the corneal disc became turbid. Restored activity of LCS in severe caustic damage of the anterior segment opened new therapeutic opportunities. PMID- 11852549 TI - [Retinal detachment after excimer laser surgery]. AB - The increasing number of patients with myopia who had an excimer operation arouses among retinal specialists and refraction surgeons the question whether laser operation can cause or accelerate retinal damage and initiate a motion of the retina. During the last 4 years the authors treated 5 patients with a motion of the retina who were previously subjected to excimer laser surgery on account of severe myopia. The laser procedures were implemented in different departments and then the patients were referred to us on account of complications. Poorer results and a poorer prognosis were recorded in patients with manifest degenerative changes of the periphery and with extensive rhegmatogenic manifestations. In patients without peripheral degeneration the optic functions after surgery of a motion were practically unrestricted. The incidence of rhegmatogenic changes and vitreal traction is according to the authors a contraindication for excimer procedures and they recommend to refer these patients to a vitreoretinal department. PMID- 11852550 TI - [Cataract surgery using silicone oil and its elimination via posterior capsulorhexis]. AB - Elimination of silicone oil after a previous vitreoretinal operation is a routine procedure. This procedure is frequently associated with a complicated cataract operation. The standard procedure of oil elimination in one or two stages assumes repeated sclerotomy. Contemporary findings and the authors' experience indicate the risk of the site of sclerotomy. The authors draw attention to a modification of silicone oil elimination while operating the cataract by posterior capsulorhexy which makes it possible to remove the intraocular tamponade without sclerotomy and to implement at the same time peroperative control of the posterior pole. the technique of elimination of silicone via posterior capsulorhexy makes a perfect peroperative check-up of the state of the posterior eye possible it makes it possible to leave the silicone in the eye if it is necessary to prolong the tamponade of the retina it reduces the risk of complications of further sclerotomy it makes reconstruction of the eye by a single operation possible and reduces thus the traumatization of the eye and the patient. PMID- 11852551 TI - [Screening for diabetic retinopathy in the Czech Republic--guideline]. AB - Diabetic retinopathy (DR) and diabetic maculopathy (DMP) are the most frequent and most serious complications of diabetes mellitus (DM). At present the adverse development of these microvascular complications of DM can be very successfully delayed by available therapeutic methods. The success of the therapeutic procedure depends in particular on early treatment of DR and DMP by laser coagulation and/or pars plana vitrectomy, which presumes detection of early stages of the disease. The authors evaluate the priorities of DR screening, the final effect of which is a decline of blindness caused by DM. Conditions for screening of diabetic eye disease are the diagnosis of DR by simple and safe procedures, classification of DR on the basis of the dynamics of retinal changes and standards of treatment. An integral part of the ophthalmological screening programme is professional collaboration of ophthalmologists, diabetologists, general practitioners and specialists in internist medicine. Successful screening of DR makes it possible to involve the subject in preventive and therapeutic care, education of the patient, regular lifelong follow up and early treatment of DR and DMP. By the method of screening of DR we reduce the risk of a decline of visual acuity and prevent severe functional losses as a result of diabetic ophthalmological complications. PMID- 11852552 TI - [Cryotherapy and photocoagulation in the treatment of retinopathy of prematurity]. AB - In a retrospective clinical trial is compared efficacy of transscleral cryotherapy versus diode laser photocoagulation in the treatment of threshold stage 3 retinopathy of prematurity (ROP) in zone II-III. 40 eyes were treated with cryotherapy and 32 eyes were treated with diode laser photocoagulation. All children have been followed up for 5 to 9 years after treatment. The long-term best-corrected visual acuity (BCVA) and refractive errors were evaluated. BCVA from 6/6 to 6/18 in last examination had 53% of eyes with cryotherapy and 56% of eyes with photocoagulation. The difference was not statistically significant (P = 0.094). Myopia over -6.0 D were at 32.5% of eyes with cryotherapy and at 22% of eyes with photocoagulation. This difference was statistically significant (P < 0.05). In the second part of this study is evaluated the long-term visual acuity and structural outcome in Zone 1 ROP (Very posterior Zone 1 disease, rush disease). 20 eyes with cryotherapy and 24 eyes with photocoagulation of this severe form ROP were followed up for 5 to 11 years. BCVA of 6/36 and better had 30% eyes of cryotherapy and 37.5% eyes of laser photocoagulation (P < 0.05). Unfavourable structural fundus outcomes had 55% eyes of cryotherapy and 33% eyes of photocoagulation (P < 0.05). These differences were statistically significant. Our results support the long-term efficacy and safety of cryotherapy and diode laser photocoagulation in the treatment of threshold 3 stage ROP. Visual acuity results suggest that diode laser photocoagulation is as effective as cryotherapy in treatment of this ROP stage. In the group of posterior Zone 1 ROP was achieved better visual acuity results and reduced unfavourable structural outcomes with diode laser photocoagulation as compared to cryotherapy. PMID- 11852553 TI - [Effect of early surgery in essential infantile esotropia on the quality of binocular vision]. AB - In a retrospective study of 397 children operated in the course of 10 years (1985 1995) on account of essential infantile esotropia the authors evaluate the effect of early surgery implemented before the age of two years on the quality of binocular vision as compared with a later operation. The group of children was divided into three sub-groups. Group A comprised 75 children with the operation during the first six months of life (mean 3.8 months), sub-group B 194 children with the operation at the age of 6-24 months and in group C 128 children operated at the age of 2-6 years (mean 3.56 years). In group A binocular vision was recorded in 80% children (15% superposition, 60% fusion, 5% stereopsy). In groups B binocular vision was recorded in 76% children (18% superposition, 50% fusion, 8% stereopsy). In group C simple binocular vision in the form of superposition was present in 24% and fusion only in 21% children. The results of binocular vision after surgery of essential infantile esotropia are in favour of early surgery, preferably by the age of 6 months, not later than at the age of 2 years. An essential part of comprehensive treatment is active and positive pleoptic and orthoptic care incl. supplementary surgical correction of residual horizontal or vertical deviations. Early surgery of an adequate extent with a safeguarded parallel position of the eyes implies in the long run more frequent achievement of a higher quality of binocular vision incl. stereopsy. PMID- 11852554 TI - [Transmission electron microscopy of the pupillary membranes in neovascular glaucoma and chronic hemophthalmos]. AB - The authors examined using transmission electron microscopy the pupillary membrane in neovascular glaucoma. The membrane was formed by connective tissue where fibroblasts, pigment cells and collagen fibres predominated. The high cell content suggested a high metabolic and growth activity. PMID- 11852555 TI - [Intraocular pressure in phacoemulsification]. AB - The authors investigated changes of the intraocular pressure after non complicated phacoemulsification with implantation of an intraocular lens of different types (OMMA, silicone, Acrysof) in 40 patients. From the results it is obvious that in some patients already 4 hours after surgery the intraocular pressure rises to an average of 31 +/- 17 mm Hg. The magnitude of the intraocular pressure was not influenced by the size of the surgical wound nor the type of intraocular lens. In values below 30 mm Hg the authors recommend merely follow up of the patients as they assume that these values have a positive impact on closure of the tunnel incision. In the differential diagnosis it is important to consider the post-operative inflammatory reaction, the absorption reaction of residual viscoelastic material, pupillary block while the lenticular capsule is intact and undiagnosed glaucoma. In values above 30 mm Hg the authors recommend administration of beta-blockers, Diluran and antiphlogistics. PMID- 11852556 TI - [Treatment of central serous chorioretinopathy--personal experience]. AB - On a group of 18 eyes the authors evaluate their experience with medicamentous and laser therapy of central serous chorioretinopathy. They report unsatisfactory results in the group of patients treated with non-steroid antiphlogistics, resorbents, vasoprotective agents, vitamins, where they observed adherence of ablated neuroepithelium in 4 eyes on average after 5.5 weeks. But in 7 similarly treated eyes adherence of the ablation did not occur even after 4 months. The authors confirm the value of direct laser photocoagulation for reducing the period of ablation in central serous chorioretinopathy. In all 11 photocoagulations of treated eyes the ablation adhered on average after 5.2 weeks. The authors maintain that they achieved better final visual acuity and greater improvement of visual acuity in patients where the ablation period of the neuroepithelium was shorter than in patients with a prolonged course of ablation. PMID- 11852557 TI - [Meteorotropy of eye diseases]. AB - Based on extensive material of patients with infectious eye diseases in southern Tunisia, North Africa, using mathematical and statistical calculations, the author provided evidence of their seasonal rhythm and confirmed thus folk traditions and old statements on trachoma. One year, special from the meteorological aspect, proved that these diseases are governed by meteorotropy. PMID- 11852558 TI - [Central serous chorioretinopathy. Etiopathogenesis, clinical picture, diagnosis. Part I]. PMID- 11852559 TI - [An insoluble colored substrate for dextranase assay]. AB - An assay of dextranase (EC 3.2.1.11) was developed by using Sephadex G-200 coupled with Remazol Brilliant Blue (RBB) as an insoluble substrate. The assay procedure included incubation of suspension of the colored substrate in buffer containing enzyme under study, removal of residual insoluble substrate, and measurement of the absorbance of supernatant fluid containing colored soluble hydrolysis products at 595 nm. The procedure was examined in the screening of dextranase-forming bacilli from the microbial collection of the Institute of Biology, Ufa Research Center, RAS. PMID- 11852560 TI - [Development of enzyme immunoassay for the herbicide chlorsulfuron]. AB - Antibodies against the herbicide chlorsulfuron have been raised and characterized. Enzyme immunoassays (EIAs) for chlorsulfuron, involving labeled antigen or labeled antibodies, have been developed. The kinetics of antigen antibody interactions in the EIA systems developed has been studied. Both systems exhibit equal sensitivity (1 ng/ml). The values of the coefficient of variation (CV), determined within the range of chlorsulfuron concentrations 1-100 ng/ml to be measured by the systems, are not in excess of 8%. The possibility of using glucose oxidase as a label in EIAs for chlorsulfuron has been demonstrated. Lack of cross-reactivity with a series of sulfonyl- and arylurea derivatives and triazines makes it possible to recommend the EIA systems developed for chlorsulfuron determination in the environment. PMID- 11852562 TI - [Restriction endonuclease Sst 12I from a strain of Streptomyces sp. recognizing the nucleotide sequence 5'-CTGCAG-3']. AB - A new restriction endonuclease Sst12I belonging to the II type and recognizing the sequence 5'-CTGCAG-3' was isolated from the bacterial strain Streptomyces sp. St-12. The enzyme hydrolyzes DNA between adenine and guanine residues; thus, it is a true isoschizomer of restrictase PstI. In contrast to PstI, the restriction endonuclease Sst12I hydrolyses DNA both at 37 degrees and 55 degrees C and remains active after long-term storage. PMID- 11852561 TI - [Testing and isolation of high-purity restriction endonucleases]. AB - A new method of testing restriction nucleases is proposed. This method is based on high-temperature treatment of crude cell extracts. Disrupted cells were heated at 50-60 degrees C, centrifuged, and assayed for restrictases. This method provides the opportunity for screening new enzymes in microbial strains enriched with nonspecific restrictases. High-temperature treatment of cell extracts of certain producers reduces the number of steps of the procedure used for isolating high-purity restrictases; the resulting preparations are capable of maintaining high enzymatic activity during long-term storage. It was shown that high temperature treatment can be applied not only to thermophilic but also to mesophilic strains of microorganisms of different taxa. PMID- 11852563 TI - [Development of biosensors for phenol determination from bacteria found in petroleum fields of West Siberia]. AB - Nine Gram-negative bacterial strains, selected from 300 strains isolated from soils of the West Siberian petroliferous basin and growing on oil and oil products, consume phenol as a single carbon and energy source. The strains were used for the development of a sensor bioreceptor. The most active 32-I strain was shown to bear a plasmid responsible for phenol degradation. The plasmid-free derivative of this strain, 32-I-1, did not grow on phenol. The possibility of creating a model biosensor for phenol based on the plasmid-containing 32-I strain is considered. The detection limit for phenol was 5 microM. The optimum conditions for the sensor operation are: pH 7.4, 35 degrees C, and operation time 30 h. PMID- 11852564 TI - [Selection of ergot alkaloid producers by induced mutagenesis]. AB - Using the induced mutagenesis technique, A series of genetically modified Claviceps sp. VKM F-2609 strains that display high levels of agroclavine and elymoclavine synthesis were selected by induced mutagenesis. Compared to the parent strain, c106 displayed a 40-fold higher level of agroclavine synthesis, and c66 displayed an eightfold higher level of elymoclavine synthesis. The levels of synthesis of other alkaloids were decreased in these strains. The effects of various carbohydrates on the strain growth and ergot alkaloid biosynthesis was then investigated in both the parent strain and c106. The largest amount of agroclavine was synthesized by c106 strain growing on a medium with maltose. PMID- 11852565 TI - [Factors contributing to roquefortine yield variability during cultivation of penicillium roquefortii]. AB - Variability in the roquefortine yield was shown to be associated with its consumption by the mycelium during isolation of the end product, which depended on temperature, time of culture liquid storage, and biomass concentration. This was also related to the presence in chloroform of chlorocarbonic acid ethyl ester that reacted with roquefortine. PMID- 11852566 TI - [Stability of glucose 6-phosphate dehydrogenase complexed with its substrate and/or cofactor in aqueous and micellar environment]. AB - Inactivation of glucose 6-phosphate dehydrogenase (G6PDH) complexed with its substrate, glucose 6-phosphate (GP), and/or cofactor, NADP+, has been studied within the range 20-40 degrees C in three media: (a) 0.04 M NaOH-glycine buffer (pH 9.1); (b) Aerosol OT (AOT) reversed micelles in octane; and (c) Triton X-100 micelles in octane supplemented with 10% hexanol. The enzyme inactivation was characterized quantitatively by first order rate constants, kin (s-1). In the case of G6PDH-NADP+ complexes, the values of kin were independent of the initial concentrations of G6PDH, either in aqueous medium or AOT micelles. The values of kin for the complex G6PDH-GP were inversely related to the initial concentration of the enzyme, in both aqueous and micellar media. When inactivation of both complexes were studied in AOT micelles, minimum values of kin corresponded to the degree of hydration W0 = 16.7; at W0 > 16.7 and W0 < 16.7, kin increased. Within the range 20-40 degrees C, the values of kin measured for both complexes in aqueous medium were significantly lower than those measured in AOT micelles. Temperature dependences of kin were characterized by inflections in Arrhenius plots, which corresponded, depending on the medium, to certain temperatures from 33.6 degrees C to 40 degrees C. In all media studied, NADP+ complexes of the enzyme exhibited higher stability than their GP counterparts. The parameters of G6PDH and G6PDH-NADP+ melting, measured by differential scanning microcalorimetry (maximum temperature and half-width of the transition, enthalpy of denaturation, and van't Hoff enthalpy), provided unequivocal evidence of the higher stability of the complex as compared to that of the enzyme. In addition, this approach demonstrated that G6PDH undergoes destabilization in AOT micelles. PMID- 11852568 TI - [Effect of phytohormones on the protein-synthesizing ability of rauwolfia serpentina benth. tissue culture]. AB - The accumulation of intracellular protein by the callus culture of Rauwolfia serpentina Benth. was studied in a standard or phytohormone-containing medium. Changes in the concentration of total protein in cells induced by indolylacetic and naphthylacetic acids were shown to be associated with the effects of these phytohormones on the biosynthesis and degradation of intracellular protein. PMID- 11852567 TI - [The antiviral activity of chitosan (review)]. AB - Data on the inhibitory effect of chitosan on viral infections in animals, plants, and microorganisms are reviewed. The effects of the physicochemical parameters and structure of chitosan on its antiviral activity are analyzed. Possible mechanisms of the inhibitory effect of chitosan on viral infections are discussed. PMID- 11852569 TI - [Effect of arsenic on bacterial growth and plasma membrane atpase activity]. AB - The effects of arsenic in the forms of arsenite and arsenate on bacterial growth and plasma membranes ATPase activity of was studied. Correlation of The rate of ATP hydrolysis was found to be correlated with bacterial resistance to toxic arsenic ions. Detoxification of arsenate by resistant cultures of bacteria was suggested to be related with an increase in bacterial ATPase activity and the degree of ATPase mobilization. PMID- 11852570 TI - [Consumption of organic carbon sources and biosynthesis of lactic acid by the photosynthetic bacterium Rhodobacter sp. D-4]. AB - Nonsulfur photosynthetic purple bacteria isolated from the Dzhermuk mineral springs (Armenia) were grown on sugar-containing media and found to be capable of synthesizing L(+)-lactic acid. Various organic compounds were tested as possible sole sources of carbon and an electron donors required to support bacterial growth and biosynthesis of lactic acid under various growth conditions. PMID- 11852571 TI - [Physicochemical properties of melanins produced by Inonotus obliquus("chagi") in the nature and the cultivated fungus]. AB - Physicochemical properties of pigments isolated from the naturally occurring sterile form of Inonotus obliquus (Fr.) Pil. known as Chagi and comprising the major constituent of the medicine befungin were compared with those of melanins synthesized by this fungus in the culture in order to develop a new medicine. Elemental and functional group analyses, as well as UV-visible, IR, and EPR spectra, and thermolysis studies revealed structural differences in these pigments and allowed for assignment of the naturally produced melanin to allomelanins, whereas that of cultivated fungus was assigned to eumelanins. PMID- 11852572 TI - [Survival of Rhizobium in monoculture and binary population with Rhizosphere bacteria]. AB - The survival of pure cultures of Rhizobium leguminosarum bv. pisum and Rhizobium trifolii and their interaction with associative diazotrophic and phosphate mobilizing bacteria after inoculation of sterile soil were studied. The viable heterotypical diazotrophic and rhizobial phosphate-mobilizing association was shown to be formed whose efficiency was 14% (clover) and 28% (pea) higher compared to monorhizobial inoculates. PMID- 11852573 TI - [Determination of the content and degree of esterification of uronic acids in plant tissues and products of their processing]. AB - A method for quantitative determination of uronic acids and the degree of esterification of their carboxyl groups in plant tissues and products of their processing is described. The method involves determination of the difference between the concentrations of Cu2+ in solution before and after interaction of copper with the substance in question. Copper was determined spectrophotometrically in the form of copper-ammonium complex. PMID- 11852574 TI - [Isolation and characterization of holocellulose from Alfalfa]. AB - Holocellulose isolated from the aerial parts of alfalfa (Medicago sativa) contains a polysaccharide complex of cellulose and hemicelluloses, the major structural components of cell walls. Holocellulose is highly hydrophilic and has a dense biopolymer packing. The carboxylic groups of hemicelluloses and cellulose determines the ability of holocellulose to adsorb polyvalent metal cations. PMID- 11852575 TI - [Volatile metabolites and external CO2 exchange of wheat cenoses under optimal conditions and thermal stress]. AB - The effects of elevated temperature (35 and 45 degrees C) on photosynthesis, respiration, and both the qualitative and quantitative compositions of volatile emissions (VE) of wheat (Triticum aestuvum L. cultivar 232) cenoses at light intensities of 70, 150, or 240 W/m2 of photosynthetically available radiation (PAR) were studied. At a PAR of 240 W/m2, the thermal stabilities of photosynthesis and respiration increased at 35 degrees C and decreased at 45 degrees C. Elevated temperatures nonuniformly changed the rates and direction of VE syntheses. In this process, the highest increase in VE evolution was observed at 70 W/m2; the lowest, at 240 W/m2 and 35 degrees C. In addition, the concentrations and composition of VE during the repair period differed from the initial values. PMID- 11852577 TI - A storm to remember. PMID- 11852578 TI - Flying lessons. Aircraft crew resource management as a model for airway resource management in the field. PMID- 11852576 TI - [Autoantibodies to human thyroid peroxidase in immunoassay and immunoaffinity chromatography]. AB - A biochemical system of radioimmunoassay of human thyroid peroxidase (TPO) with the use of specific autoantibodies was developed for the first time. This system includes lyophilized preparations of human autoantibodies to TPO, radioiodinated TPO, standards prepared from pure TPO, and the solid-phase protein A as a precipitating agent. An analytical system was used to study the TPO distribution in fractions of thyroid tissue homogenate. Differences in TPO concentrations in mitochondria, microsomes, and supernatant fluid depending on thyroid pathology and tissue storage conditions were found. The behavior of immobilized anti-TPO autoantibodies in immunoaffinity chromatography of this microsomal antigen was studied. PMID- 11852579 TI - Information integration: Virginia crash response system. PMID- 11852580 TI - Be prepared: Jamboree medical providers take Boy Scouts' motto to heart. PMID- 11852581 TI - [Physiology of microflora in the digestive tract]. AB - The microflora of the digestive tract is a complex microbial ecosystem, well balanced, which in an aboral direction undergoes specific changes as to the ratio of aerobic and anaerobic microorganisms the functions of which supplement each other--the aerobes ensure for the whole ecosystem the scavenger effect. The microbial profile of the digestive tract is typical by the absence of anaerobic microorganisms in the stomach and conversely their absolute predominance in the distal colon. The basic physiological functions of the microflora of the digestive tract can be characterized as follows: 1. microbial barrier against pathogens and potential pathogens, 2. formation of products of the microflora and their influence on the blood supply of the intestinal mucosa and peristaltics, 3. stimulation of the immune system in the gut, 4. reduction of bacterial translocation, 5. production of vitamins. To this problem in the world literature, contrary to ours, deserved attention has been paid for many years. Knowledge of the problem opens the door to expedient manipulation with the microbial flora of the digestive tract by the use of diet or prebiotics, probiotics as well as antibiotic treatment. The author presents a summary of basic findings which developed on the subject of physiological microflora of the digestive tract successively up to their present shape. PMID- 11852582 TI - [Zinc and copper in diseases of the digestive tract] ]. AB - The author mentions the basic metabolic functions of zinc and copper in the human organism. She emphasizes the relationship of serious diseases, congenital and acquired, associated with deficiency or excess of these mineral trace elements. As to inborn diseases, we encounter most frequently Wilson's disease in impaired copper metabolism, rarely morbus Danbolt in congenital impairment of zinc absorption. In practice we are faced in particular with secondary causes of impaired zinc or copper levels, e.g. in chronic diarrhoeal conditions, coeliac disease, chronic pancreatitis and others. PMID- 11852583 TI - [Pathophysiology of hepatic fibrosis]. AB - Hepatic fibrosis is a common sequel to most forms of chronic liver disease and an essential component in the development of cirrhosis. Basic and clinical scientists have increasingly realized the importance of all elements of the liver in fibrogenesis, as well as of structural-functional relationships between liver cells and matrix components of the liver. Important is the recognition that hepatic stellate cells play a central role based on their ability to undergo activation following liver injury of any cause. The study brings about the newest information on pathophysiology of hepatic fibrosis. On the basis of better knowledge of the pathophysiology of fibrogenesis the development of new therapeutic approaches will become possible. PMID- 11852584 TI - [Big endothelin and chronic heart failure]. AB - In a group of 124 patients the authors investigated the importance of assessment of plasma levels of big endothelin and endothelin 1 in patients with chronic heart failure as compared with other currently used non-invasive parameters. A six fold increase of plasma levels of both substances was found in patients in functional class NYHA IV as compared with patients in class NYHA II-III. But even patients in the milder stage of NYHA had twice as high values as compared with the standard of the healthy population. Similarly patients with interstitial pulmonary oedema had a twice as high level of both parameters as compared with patients who had a normal finding on X-ray or merely a redistribution of the pulmonary vascularization. The sensitivity of assessment of plasma levels is such that this examination could become part of the basic diagnosis. PMID- 11852585 TI - [Pulmonary alveolar proteinosis and its treatment with total pulmonary lavage in the Czech Republic]. AB - Pulmonary alveolar proteinosis is a rare disease characterized by accumulation of a large amount of both components of surfactant, proteins and phospholipids in alveoli and terminal bronchioles. This cumulation may lead to impaired gas exchange and result in respiratory insufficiency. The disease may develop in neonatal age or later in adult age. Two congenital forms are known. The disease occurs however also in people with neoplastic disease, during cytostatic treatment, during inhalation of inorganic dusts. A dominant symptom is progressing dyspnoea. Asymptomatic forms exist also. The most effective therapeutic method is overall pulmonary lavage. The authors implemented in 1999 the first overall pulmonary lavage in the Czech Republic in the General Faculty Hospital in Prague in a 60-year-old female patient. PMID- 11852586 TI - [Cor thyreotoxicum. Part I--new findings about its etiopathogenesis and diagnosis. Overview of problems based on 35 years' experience]. AB - In the seventies thyrotoxic heart accounted for 3% of all hospitalized cardiac patients and was found on average in 30% of all cases of hyperthyroidism. It presented most frequently by tachyfibrillation and resistant cardiac decompensation. It affected men four times as frequently as women. The incidence correlated with age, toxic nodose goitre, but its development did not correlate with concurrent thyrotoxic rhizomyelic myopathy nor with the extent of deviation of thyroid laboratory parameters (T4, T3, indexes FT4). At present the incidence of thyrotoxic heart declined due to early detection and more adequate diagnosis and treatment of hyperthyroidism, as well as due to the decline of oligosymptomatic toxic nodose goitres even in old age due to preventive iodization of table salt. However, there was an increase of hyperthyroidism induced by amiodarone and other iodine preparations (X-ray contrast materials) associated with primary heart disease and arrhythmias. (Up to 2% of amiodarone treated patients). The ratio of so-called real subclinical thyrotoxicoses in the development of thyrotoxic heart is negligible. Isolated reduction of TSH in hospital screening is a frequent finding but is conditioned most frequently by: a) the 1st stage of the low thyroxin syndrome, b) the 1st stage of subacute thyroiditis, c) the influence of various drugs (iodine preparations, overdosage of T4 substitution, pharmacotherapy with glucocorticoids, dopamine etc.), d) methodical artefacts, e) natural pulsed secretion of TSH etc. Hospital screening of hyperthyroidism and thyrotoxic heart even in older people above 60 years by T4 and/or TSH (2nd generation equipment) is not effective because it is detected in 20% of current hospital admissions and in 60% of those admitted to intensive care unitpathologic values of T4 and/or TSH most frequently without non-thyroid causes (stages of the low thyroxin syndrome) are recorded. This hospital screening has a satisfactory sensitivity but low specificity and in a large number of people calls for further diagnostic steps. Therefore it is more suitable only after clinical examination of the patient to confirm suspected hyperthyroidism to examine FT4 and TSH (IRMA 3rd generation) or possibly supplement FT3 and other aimed tests. PMID- 11852587 TI - [Chronic myeloid leukemia as a model for modern noncytostatic therapy of malignant diseases. From Virchow to STI 571]. AB - Chronic myelogenous leukemia (CML) has been described in the middle of the 19th century. It took further 100 years to the time of the discovery of the first cytotoxic drugs. Ph1 chromosome, the cytogenetic hallmark of CML, has been described in 1960, however, the majority of important discoveries on the molecular level has been done in the 1980s' and later. The chromosomal translocation t(9;22)(q34;q11) is absolutely crucial for CML and leads to the chimeric gene and protein BCR-ABL. The ABL protein possesses the tyrosine kinase activity, which is deregulated in the BCR-ABL protein, leading to the cascade of further pathological processes in the leukemic cells and causing eventually their malignant transformation. In the 1990s, the drug STI 571 has been synthesized, which is a potent inhibitor of the ABL tyrosine kinase activity. The data so far published concerning clinical effects of STI 571 are remarkable and very encouraging. The drug seems to be very safe and has only limited toxicity. It must be emphasized, however, that STI 571 has been clinically tested only since 1998. Moreover, from the mechanisms of action it is clear, that STI 571 influences the secondary, not the primary cause of the CML. Nevertheless, it is without any doubts that the development of new anti-cancer drugs will be done on this non-cytotoxic principle. PMID- 11852588 TI - [Visceral panniculitis in occult pancreatic carcinoma. Case report from clinico pathologic archives]. AB - The authors describe a 75-year-old man with acute abdominal disease leading to lethal prerenal failure. Necropsy revealed massive visceral panniculitis with a small adenocarcinoma of the head of the pancreas. The authors assume that a very rare, early paraneoplastic syndrome was involved. PMID- 11852589 TI - [Dysfunction of a valve prosthesis]. AB - The authors describe the case of a haemodynamically unstable patient with dysfunction of a valvular prosthesis which was resolved by emergency surgery. In the discussion they deal with two basic causes of these dysfunctions and their possible solutions. PMID- 11852590 TI - [Serious side-effects of interferon alfa and ribavirin combination therapy in patients with chronic hepatitis C]. AB - At present standard treatment of patients with chronic hepatitis C is combined therapy with interferon-alpha and ribavirin which so far gives the best results. It is recommended in patients with resistance to interferon or a relapse after termination of treatment by interferon alone as well as in hitherto untreated patients. Combined treatment produces, however, not only common side-effects which are caused by interferon as well as ribavirin but rarely also serious side effects produced by combined treatment which call for premature termination of therapy. The authors present an account on two patients who had serious side effects during combined treatment: the first one was an intravenous drug addict who developed psychosis during combined treatment, the second one a female patient with cirrhosis and incipient portal hypertension who developed after nine months of combined treatment a severe biochemical relapse with jaundice, and treatment was also terminated. Both serious complications after treatment receded and in the first patient a sustained response to antiviral therapy persists for more than one year. PMID- 11852591 TI - [Franz Kafka in the Tatra Mountains]. PMID- 11852592 TI - [History of treatment of essential hypertension--part I]. PMID- 11852593 TI - [Reliability and validition of the Slovak modified version of the Stanford Health Assessment Questionnaire using the functional disability index in patients with rheumatoid arthritis]. AB - BACKGROUND: Functional disability is one of the most important consequences of RA in the patient's daily life. The HAQ has been widely used in its self administered form for the assessment of disability. A sensitive and valid instrument is needed for a Slovak population with RA. OBJECTIVE: To evaluate reliability and validity of the Slovak version of the HAQ in patients with RA and to explore relationships between HAQ score and disease activity and to provide information concerning utilization of this instrument in clinical practice. METHODS: In preparing the Slovak version of the HAQ careful attention has been paid to the translation, in terms of linguistic and conceptual equivalence, in order to preserve the original purpose of the instrument. The wording of some items required adaptation to the current activities of daily living and Slovak lifestyle. The sample consisted of 160 RA-patients, out of which 135 were women and 25 were men. The inclusion criteria were the following: age from 20 to 70 years at the onset of the study, diagnosis of RA according to the ARA criteria. The exclusion criteria were the presence of another serious disease or very disabling RA (stage IV of the Steinbrocker's classification). To analyze the data t-test, correlations, one-way analysis of variance (ANOVA), and principal component analysis (PCA) available in the SPSS/PC+ statistical package were used. RESULTS: The Cronbach's coefficient of reliability alpha for the HAQ total scale was 0.94. The results of PCA showed that the 20 HAQ items loaded on four components for which the eigen values were greater than 1, accounting for 70% of overall interpersonal variability. Orthogonal varimax rotation of the principal components provided factor loadings reflecting the eight dimensions within the HAQ. Validity of the HAQ was examined further by means of known-groups technique. The HAQ was found to be sensitive to differentiate between the Steinbrocker's functional capacity groups, as well as between males and females. Moreover, significant correlations (p < or = 0.01) were found between the HAQ and the C reactive protein, the ESR, the NHP-pain, the Ritchie articular index, the Steinbrocker's functional capacity (r = 0.31-0.62) and disease duration (p < or = 0.05, r = 0.17). CONCLUSION: The results of the current investigation provide support for reliability and construct validity of the Slovak version of the HAQ in patients with RA. The HAQ has sufficient discriminant ability. The index disability can be used as an criterion of severity of RA, as a criterion of effectiveness in therapeutical trials for patient stratification of the Slovak population with RA. PMID- 11852594 TI - Simulation approaches to ion channel structure-function relationships. PMID- 11852595 TI - Dynamics of biochemical and biophysical reactions: insight from computer simulations. PMID- 11852596 TI - Partnering to promote and enhance medication safety. AB - Addressing medication errors and supporting medication safety has become an important quality initiative. But are hospitals and health systems using the best practices that are available today to order, dispense, and monitor medications for patients. Many are finding that they can achieve a stronger medication safety program through collaboration. This issue looks at how hospitals and health systems are partnering--with other hospitals and health systems locally and nationally, health plans, health care purchasing groups, universities, vendors, and even patients--to reduce the likelihood of medication errors. PMID- 11852598 TI - Hand-held computers in medicine. PMID- 11852597 TI - What patients want to know about their medications. Focus group study of patient and clinician perspectives. AB - OBJECTIVE: To describe what patients want to know about their medications and how they currently access information. To describe how physicians and pharmacists respond to patients' information needs. To use patients', physicians', and pharmacists' feedback to develop evidence-based treatment information sheets. DESIGN: Qualitative study using focus groups and a grounded-theory approach. SETTING: Three regions of Canada (British Columbia, Nova Scotia, and Ontario). PARTICIPANTS: Eighty-eight patients, 27 physicians, and 35 pharmacists each took part in one of 19 focus groups. METHOD: Purposeful and convenience sampling was used. A trained facilitator used a semistructured interview guide to conduct the focus groups. Analysis was completed by at least two research-team members. MAIN FINDINGS: Patients wanted both general and specific information when considering medication treatments. They wanted basic information about the medical condition being treated and specific information about side effects, duration of treatment, and range of available treatment options. Physicians and pharmacists questioned the amount of side-effect and safety information patients wanted and thought that too much information might deter patients from taking their medications. Patients, physicians, and pharmacists supported the use of evidence-based treatment information sheets. CONCLUSION: Patients and clinicians each appear to have a different understanding of what and how much information patients should receive about medications. Feedback from patients can be used to develop patient oriented treatment information. PMID- 11852599 TI - Managing adults with urinary incontinence. Clinical practice guidelines. PMID- 11852600 TI - Gaps in mental health services for seniors. What can we do about them? PMID- 11852601 TI - Residents' page. Requiem for Mr Bojangles. PMID- 11852602 TI - Hypothesis: the research page. One researcher, two researcher, three researcher, four. Five researcher, six researcher, seven researcher, more! PMID- 11852603 TI - Get active about physical activity. Ask, advise, assist: get your patients moving. PMID- 11852604 TI - Urinary incontinence. Keeping family physicians involved in care. PMID- 11852605 TI - New face, new vision. PMID- 11852606 TI - How reliable is this test? PMID- 11852607 TI - Organized stroke care is emerging in Ontario. PMID- 11852609 TI - Ophthaproblem. Prepapillary vascular loops. PMID- 11852608 TI - Drinking alcohol while breastfeeding. Will it harm my baby? AB - QUESTION: I recently delivered a healthy, full-term baby and am now breastfeeding exclusively. I abstained from drinking alcohol during my entire pregnancy and am wondering if drinking alcohol now would harm my nursing baby. ANSWER: Nursing mothers who choose to drink alcohol during the postpartum period should carefully plan a breastfeeding schedule by storing milk before drinking and waiting for complete elimination of alcohol from their breast milk after drinking. Motherisk has created an algorithm to estimate how long it takes to eliminate alcohol from breast milk. PMID- 11852610 TI - Practice tips. Putting the "Palm" into practice. PMID- 11852611 TI - Just the berries. Diagnosing and managing group A streptococcus pharyngitis. PMID- 11852612 TI - Should patients who have not had a cardiac event take ASA to prevent one? PMID- 11852613 TI - Asymptomatic bacteriuria during pregnancy. Rapid answers using the Cochrane Library. PMID- 11852614 TI - Physical activity to prevent cardiovascular disease. How much is enough? AB - OBJECTIVE: To review the role of physical activity in primary prevention of cardiovascular (CV) diseases with particular attention to the intensity and amount of physical activity needed to benefit health. QUALITY OF EVIDENCE: MEDLINE was searched for articles published in the indexed English literature from January 1991 to December 2000 using key words related to physical activity (e.g., exercise, physical fitness), CV and coronary artery disease (CAD) risk factors (e.g., diabetes, hypertension, hyperlipidemia, obesity). Findings were supplemented by consensus documents and other published literature. Most articles described prospective observational studies. MAIN MESSAGE: Clear evidence indicates an inverse linear dose response between amount of physical activity and all-cause mortality, total CV disease, and CAD incidence and mortality. The minimal effective dose is unclear, but physical activity that results in energy expenditure of approximately 4200 kJ.week-1 appears to be associated with substantial benefits. Physical activity need not be vigorous to benefit health. CONCLUSION: Moderate activity, such as brisk walking for 30 to 60 minutes a day most days of the week, is associated with significant reductions in the incidence and mortality of CV disease. PMID- 11852615 TI - Does counseling help patients get active? Systematic review of the literature. AB - OBJECTIVE: To determine the effect of counseling patients to become more physically active. DATA SOURCES: PubMed was searched for articles during the past 30 years on physicians promoting physical activity. Identified studies were cross referenced, and experts were consulted for additional articles. STUDY SELECTION: Thirteen articles described primary care counseling on exercise. Six studies were randomized controlled trials (RCTs); seven were quasi-experimental designs. Three of the four RCTs and three of the five quasi-experimental studies were short term (4 weeks to 2 months); the remaining three trials lasted longer than 6 months. Most studies used strategies to address stage of change. SYNTHESIS: Outcome measures included adoption of physical activity, stage of change, and change in physical activity level. Most studies found positive relationships between counseling and these outcomes. No reliable evaluation instruments were found, nor was the long-term effect of interventions established. CONCLUSION: Interventions that included written materials for patients, considered behaviour change strategies, and provided training and materials for physicians were effective at increasing levels of physical activity. New strategies that involve measuring and prescribing specific amounts of exercise might also improve fitness levels and hence improve outcomes of chronic disease. Shortcomings of these studies include lack of long-term data, lack of sustaining activities for family physicians, and scant cost-efficacy analysis. PMID- 11852616 TI - Urinary incontinence in Canada. National survey of family physicians' knowledge, attitudes, and practices. AB - OBJECTIVE: To determine current knowledge, attitudes, and management of urinary incontinence among family physicians in Canada. DESIGN: Cross-sectional mailed survey. SETTING: Family physicians in Canada. PARTICIPANTS: A random sample of 1500 members of the College of Family Physicians of Canada. MAIN OUTCOME MEASURES: Self-assessed knowledge, self-reported attitudes, and rating of various tests and treatments in the investigation and management of incontinence. RESULTS: The overall unadjusted response rate was 43.3% (650/1500). Although most respondents reported that urinary incontinence was common in their practices, less than half (46.0%, 284/617) indicated that they clearly understood incontinence and just 37.9% (232/612) had an organized plan for incontinence problems. Only 35.0% (214/612) of respondents felt very comfortable dealing with incontinence. Physical examination, urodynamic studies, urinalysis, and testing blood sugar levels were all considered important investigations by more than 90% of respondents. CONCLUSION: There are wide variations in knowledge, attitudes, practices, and comfort level among family physicians dealing with urinary incontinence. PMID- 11852618 TI - Management of pneumothorax. PMID- 11852617 TI - Medical conditions, medications, and urinary incontinence. Analysis of a population-based survey. AB - OBJECTIVE: To assess associations between various medical conditions and drug treatments and reports of urinary incontinence. DESIGN: Secondary analysis of responses to the second wave of the National Population Health Survey (NPHS). Odds ratios were calculated using survey-weighted multiple logistic regression; confidence intervals were calculated using bootstrap methods. SETTING: Canadian households in all 10 provinces, as assessed by Statistics Canada's NPHS. PARTICIPANTS: From among respondents to the NPHS, the 54,920 people aged 30 years or older. MAIN OUTCOME MEASURES: Responses to the question "Do you have urinary incontinence diagnosed by a health professional?" and analysis of variables related to medical conditions and medications. RESULTS: Urinary incontinence was associated with strokes, arthritis, and back problems in both sexes. Odds ratios for incontinence were elevated among men and women who reported having asthma. Narcotics and diuretics were strongly associated with incontinence in both sexes. Psychoactive medications were associated with incontinence in women; antidepressants were associated with incontinence in men. CONCLUSION: Physicians should consider the possibility that patients with common conditions, such as arthritis, back problems, or respiratory conditions associated with coughing, might also have urinary incontinence. Physicians should also be aware that urinary incontinence might be a side effect of therapies and make relevant inquiries. Medications associated with incontinence could be changed. PMID- 11852619 TI - Silicosis and coal workers' pneumoconiosis. PMID- 11852620 TI - Dyslipidaemia in diabetes--a GP guide. PMID- 11852621 TI - A father complains about his violent son. PMID- 11852622 TI - Reducing asthma deaths. PMID- 11852623 TI - Key developments in respiratory medicine. PMID- 11852624 TI - Helping patients to stop smoking. PMID- 11852625 TI - Casebook: chronic cough. PMID- 11852626 TI - Preventing asthma deaths. PMID- 11852627 TI - Osteopenia has a differential diagnosis. PMID- 11852628 TI - Clinical Standards Board for Scotland: peer review visit. PMID- 11852629 TI - Anthrax and other microbial threats. PMID- 11852631 TI - Experience of a small British pacing centre between 1994 and 2000: some answers to the problem of low UK implantation rates. AB - Raigmore is a district general hospital offering a permanent pacemaker service to its catchment population of 233,500. This report reviews the pacemaker practice over a seven year period (1994 to 2000) and makes a comparison with the national database and other hospitals in the UK. The records of all patients receiving new implantions for the period under observation were reviewed retrospectively. Data collected included number of patients paced each year, age, sex, indications and complications. In the 84 months studied 3/71 patients received new permanent pacemakers (the mean age was 74 years and 51% were male). The most common presenting symptoms were syncope (62%), dizzy spells (24%), and heart failure (11.5%). The most common indication for permanent pacemaker insertion was complete atrioventricular block (58%) followed by sick sinus syndrome (20%). The implantation rate was 419.3/million population/year in 2000. Pacemaker modes used were appropriate and the early and late complication rates were low (2.7%). This current audit demonstrates continued growth of the service with low-complication rates and implantation rates which approach those in Western Europe and North America. PMID- 11852630 TI - Preferences for different high-energy foods in elderly medical in-patients. AB - Malnourishment is a common finding in hospitalised elderly patients. It is often addressed by the provision of nutritional supplements, in the form of sip-feeds. Patients' intake of these is frequently inadequate. We assessed the palatability of sip-feed nutritional supplements and other high-energy foods to elderly medical in-patients. Using the Lickhert Scale, 49 subjects rated the taste of a previously selected sip-feed supplement and five other high-energy foods, cheese biscuit, plain potato crisps, chocolate, cherry-flavoured cereal bar and stout beer. Subjects rated the taste of sip-feeds as favourably as all other offered foods, with the exception of the lower rated stout beer (p = 0.0001). Taste alone is unlikely to account for the poor intake of sip-feed nutritional supplements by elderly hospital patients. PMID- 11852632 TI - Diving accidents in sports divers in Orkney waters. AB - Scapa Flow in Orkney is one of the major world centres for wreck diving. Because of the geography of Orkney and the nature of the diving, it is possible to make relatively accurate estimates of the number of dives taking place. The denominator of dive activity allows the unusual opportunity of precise calculation of accident rates. In 1999, one in every 178 sports divers visiting Orkney was involved in a significant accident, in 2000 the figure was one in 102. Some of these accidents appear to have been predictable and could be avoided by better education and preparation of visiting divers. PMID- 11852633 TI - Relationship of volume of lesion to length of hospital stay and outcome at one year in stroke patients. AB - The aim of this study was to investigate the relationship between the volume of lesion (VOL) in patients with stroke and the associated length of hospital stay (LOS), as well as longer-term functional outcome. Computerised tomography (CT) scans were used to measure the volume, region and type of lesion, volume being measured by planimetry. LOS and other patient details were obtained from the Dundee Stroke Database. The total LOS was associated with the VOL on univariate analysis (p = 0.004) and after adjustment for the other variables (p = 0.006) due to a larger lesion being associated with longer stay in hospital. Patient follow up confirmed that the VOL was also highly significant when related to functional outcome measures of impairment, disability and handicap at one year, as determined by Orgogozo (p = 0.03), Barthel (p < 0.01) and Rankin scores (p < 0.01) respectively. The VOL is related to the length of stay in hospital and outcome at one year. This is of particular interest with the increasing use of thrombolysis and development of neuroprotectant agents designed to limit VOL. PMID- 11852634 TI - Rash, abdominal pain and hyponatraemia. AB - Hypertriglyceridaemia is a rare precipitant of acute pancreatitis. We present a patient with acute pancreatitis and hyponatraemia in association with severe hyperlipidaemia, predominantly hypertriglyceridaemia. The patient was successfully treated with plasma exchange therapy. PMID- 11852635 TI - Creutzfeldt-Jakob disease: the need to widen the differential diagnosis. AB - We report the case of a 76 year old woman who presented with a clinical picture initially typical of a vascular dementia. The diagnosis was revised to Creutzfeldt-Jacob Disease in the terminal stages. While Creutzfeldt-Jacob Disease is rare, one must consider it in the differential diagnosis as it can mimic other dementias particularly in the chronic form which we describe here. PMID- 11852637 TI - Quantitative structure-activity studies of insect growth regulators: XIX. Effects of substituents on the aromatic moiety of dibenzoylhydrazines on larvicidal activity against the beet armyworm Spodoptera exigua. AB - Larvicidal activity against the beet armyworm, Spodoptera exigua (Hubner), was measured after topical treatment for a series of N-tert-butyl-dibenzoylhydrazines having various substituents in the benzoyl (A-ring) moiety closer to the tert butyl group, the other benzoyl (B-ring) moiety being unsubstituted. The effects of substituents on the larvicidal activity were analyzed using the classical quantitative structure-activity relationship (QSAR) procedure. Introduction of hydrophobic substituents with a small volume into any position was favourable to activity. The existence of electron-withdrawing substituents at ortho positions was also favourable to activity. For multi-substituted compounds, physico chemically unknown unfavourable factors were suggested to remain after separating common substituent effects derived from QSAR for mono-substituted analogues. With the exception of the unsubstituted compound RH-5849, the effect of substituents in the A-ring moiety on the larvicidal activity was similar to those found with the lepidopteran rice stem borer, Chilo suppressalis (Walker). The larvicidal activity of RH-5849 against S exigua was significantly lower than the value predicted from the correlation between activities against S exigua and C suppressalis. Topical treatment with piperonyl butoxide, a synergist inhibiting oxidative metabolism, slightly enhanced the larvicidal activity of RH-5849 against S exigua. PMID- 11852636 TI - New developments in insecticide resistance in the glasshouse whitefly (Trialeurodes vaporariorum) and the two-spotted spider mite (Tetranychus urticae) in the UK. AB - A recent survey of insecticide resistance in two of the most problematic pests in UK glasshouses revealed some new developments. At least some individuals in all UK samples of Trialeurodes vaporariorum that were tested resisted the insect growth regulator (IGR) buprofezin. The most strongly resistant strains were unaffected by the field application rate of this compound, and even samples from populations that had never been exposed to buprofezin contained individuals that survived the highest concentration applied (10,000 mg litre-1). The field rate of buprofezin was shown to select for resistance through vapour action alone. The benzophenylurea teflubenzuron, an unrelated IGR, was cross-resisted by buprofezin resistant individuals. There was no evidence of resistance to imidacloprid, but all T vaporariorum strains tested, regardless of origin, exhibited a high innate tolerance to nicotine, when compared with another whitefly species, Bemisia tabaci. Marked resistance to fenbutatin oxide and tebufenpyrad was found in single glasshouse populations of Tetranychus urticae, but these compounds and abamectin appeared to remain highly effective against all other strains collected. PMID- 11852638 TI - Biological properties of the novel fungicide cyazofamid against Phytophthora infestans on tomato and Pseudoperonospora cubensis on cucumber. AB - Cyazofamid (ISO proposed common name), 4-chloro-2-cyano-N,N-dimethyl-5-p tolylimidazole-1-sulfonamide is a novel fungicide exhibiting specific activity against diseases caused by Oomycetes. In tests, cyazofamid at 0.4-1.6 mg litre-1 exhibited excellent preventative activity against Phytophthora infestans on tomato and Pseudoperonospora cubensis on cucumber. Minimum inhibitory concentrations of cyazofamid against both diseases were over 63 times lower than those of mancozeb and at least 16 times lower than those of metalaxyl. Cyazofamid at 1.6-25 mg litre-1 exhibited not only preventative activity, but also stable residual activity and rainfastness. Cyazofamid at 6.3 mg litre-1 reduced zoosporangia formation of P infestans and P cubensis on host plants by 100 and 94% respectively. Cyazofamid also exhibited translaminar and curative activity. Cyazofamid has a new mode of action for fungicides and exhibits no cross resistance with other currently registered and commonly used fungicides. These properties lead to a high level control by cyazofamid in field. PMID- 11852639 TI - Predicted impact of transgenic, herbicidetolerant corn on drinking water quality in vulnerable watersheds of the mid-western USA. AB - In the intensely farmed corn-growing regions of the mid-western USA, surface waters have often been contaminated by herbicides, principally as a result of rainfall runoff occurring shortly after application of these to corn and other crops. In some vulnerable watersheds, water quality criteria for chronic human exposure through drinking water are occasionally exceeded. We selected three settings representative of vulnerable corn-region watersheds, and used the PRZM EXAMS model with the Index Reservoir scenario to predict corn herbicide concentrations in the reservoirs as a function of herbicide properties and use pattern, site characteristics and weather in the watersheds. We compared herbicide application scenarios, including broadcast surface pre-plant atrazine and alachlor applications with a glyphosate pre-plant application, scenarios in which losses of herbicides were mitigated by incorporation or banding, and scenarios in which only glyphosate or glufosinate post-emergent herbicides were used with corn genetically modified to be resistant to them. In the absence of drift, in almost all years a single runoff event dominates the input into the reservoir. As a result, annual average pesticide concentrations are highly correlated with annual maximum daily values. The modeled concentrations were generally higher than those derived from monitoring data, even for no-drift model scenarios. Because of their lower post-emergent application rates and greater soil sorptivity, glyphosate and glufosinate loads in runoff were generally one fifth to one-tenth those of atrazine and alachlor. These model results indicate that the replacement of pre-emergent corn herbicides with the post-emergent herbicides allowed by genetic modification of crops would dramatically reduce herbicide concentrations in vulnerable watersheds. Given the significantly lower chronic mammalian toxicity of these compounds, and their vulnerability to breakdown in the drinking water treatment process, risks to human populations through drinking water would also be reduced. PMID- 11852640 TI - Fumigant activity of (E)-anethole identified in Illicium verum fruit against Blattella germanica. AB - The insecticidal activities of materials derived from the fruit of star anise, Illicium verum, against adults of Blattella germanica were examined by direct contact application and fumigation methods, and compared with those of DDVP, deltamethrin and hydramethylnon. The biologically active constituent of the Illicium fruit was characterized as the phenylpropene, (E)-anethole, by spectroscopic analysis. In a filter paper diffusion test with females, (E) anethole caused 80.3% mortality at 0.159 mg cm-2 at 1 and 3 days after treatment (DAT), whereas 16.7% mortality at 3 DAT was achieved at 0.079 mg cm-2. DDVP and deltamethrin gave > 90% mortality at 0.019 mg cm-2 at 1 DAT. At 0.009 mg cm-2, DDVP and deltamethrin showed 73.3 and 60% mortality at 1 DAT, respectively, but 93.3 and 76.7% mortality at 3 DAT. Hydramethylnon exhibited 0 and 93.3% mortality at 0.159 mg cm-2 at 1 and 3 DAT, respectively, whereas 6.7% mortality at 3 DAT was observed at 0.079 mg cm-2. In a fumigation test with females, (E)-anethole was much more effective in closed cups than in open ones, indicating that the insecticidal activity of the compound was largely attributable to fumigant action. (E)-Anethole and DDVP caused 100% mortality at 0.398 and 0.051 mg cm-2 4 and 1 h after treatment, respectively. (E)-Anethole showed 46.7% mortality at 0.199 mg cm-2 at 3 DAT, whereas deltamethrin and hydramethylnon at 0.796 mg cm-2 was ineffective for 3-day period. As naturally occurring insect-control agents, the I verum fruit-derived materials described could be useful for managing populations of B germanica. PMID- 11852641 TI - Enhanced fumigant toxicity of p-cymene against Frankliniella occidentalis by simultaneous application of elevated levels of carbon dioxide. AB - The fumigant toxicity of the essential oil component p-cymene was assessed against Western Flower Thrips, Frankliniella occidentalis. F occidentalis adult females, first- and second-instar larvae and eggs were exposed for 2, 24 and 48 h to combinations of three p-cymene doses and two carbon dioxide levels (ambient, 10%). Additional experiments were conducted on F occidentalis adult females with lower carbon dioxide levels (ambient, 2%, 4%, 6%) applied in combination with p cymene. Combined applications of p-cymene and carbon dioxide were found to increase significantly the fumigant toxicity of p-cymene against both adult female and larval thrips, but not thrips eggs. An increase in exposure time also led to an increase in adult and larval mortalities in both the p-cymene alone and combined treatments. These results indicate that by combining applications of the essential oil component, p-cymene, with increased carbon dioxide, it may be possible to achieve toxicity levels similar to those of standard chemical fumigants. PMID- 11852642 TI - Susceptibility of house flies (Diptera: Muscidae) exposed to commercial insecticides on painted and unpainted plywood panels. AB - House flies were collected from dairies in New York state and the levels of resistance to commercially available insecticide formulations were measured on painted and unpainted plywood panels. Dimethoate was ineffective on all surfaces. The wettable powder permethrin formulation was more toxic than the emulsifiable concentrate formulation. The wettable powder cyfluthrin formulation was also more toxic than the recently developed liquid formulation. In general, the best house fly control was obtained on flat latex painted plywood panels and the poorest control on gloss latex painted panels. It is highly unlikely that producers obtain adequate control with dimethoate and permethrin. PMID- 11852643 TI - Photolysis of fluchloralin in aqueous methanol. AB - The photodegradation of fluchloralin by UV irradiation or sunlight in aqueous methanolic solution has been examined. In the presence of titanium dioxide five photoproducts were obtained, but only four in its absence. One photoproduct, 2, 2'-azoxy-bis(alpha,alpha,alpha-trifluoro-6-nitro-p-toluidine) is reported for the first time as a metabolite of fluchloralin. In natural sunlight the rate of degradation was higher than in UV light and titanium dioxide had almost no effect on the rate of degradation. PMID- 11852644 TI - Thermal decomposition and isomerization of cis-permethrin and beta-cypermethrin in the solid phase. AB - The stability to heart of cis-permethrin and beta-cypermethrin in the solid phase was studied and the decomposition products identified. Samples heated at 210 degrees C in an oven in the dark showed that, in the absence of potassium chlorate (the salt present in smoke-generating formulations of these pyrethroids), cis-permethrin was not isomerized, although in the presence of that salt, decomposition was greater and thermal isomerization occurred. Other salts of the type KXO3 or NaXO3, with X being halogen or nitrogen, also led to a considerable thermal isomerization. Heating the insecticides in solution in the presence of potassium chlorate did not produce isomerization in any of the solvents assayed. Salt-catalysed thermal cis-trans isomerization was also found for other pyrethroids derived from permethrinic or deltamethrinic acid but not for those derived from chrysanthemic acid. The main thermal degradation processes of cis-permethrin and beta-cypermethrin decomposition when potassium chlorate was present were cyclopropane isomerization, ester cleavage and subsequent oxidation of the resulting products. Permethrinic acid, 3-phenoxybenzyle chloride, alcohol, aldehyde and acid were identified in both cases, as well as 3-phenoxybenzyl cyanide from beta-cypermethrin. A similar decomposition pattern occurred after combustion of pyrethroid fumigant formulations. PMID- 11852645 TI - Interaction of dinotefuran and its analogues with nicotinic acetylcholine receptors of cockroach nerve cords. AB - To investigate the action of dinotefuran (MTI-446, 1-methyl-2-nitro-3-(tetrahydro 3-furylmethyl)guanidine), a recently developed insecticide, on insect nicotinic acetylcholine receptors (nAChRs), we determined the potencies of the compound and 15 analogues in inhibiting the specific binding of [3H]epibatidine (EPI), a nAChR agonist, and [3H]alpha-bungarotoxin (alpha-BGT), a competitive nAChR antagonist, to the nerve cord membranes of American cockroaches (Periplaneta americana). Racemic dinotefuran inhibited [3H]EPI binding with an IC50 of 890 nM and [3H]alpha-BGT binding with an IC50 of 36.1 microM. Scatchard analysis indicated that the dinotefuran inhibition of [3H]EPI binding was a competitive one. Slight structural modification caused a drastic reduction in potency; only four analogues were found to be equipotent to or more potent than dinotefuran. Chloropyridinyl and chlorothiazolyl neonicotinoid insecticides displayed two or three orders of magnitude higher potency than dinotefuran. There was a good correlation between the IC50 values of tested compounds obtained with [3H]EPI and those obtained with [3H]alpha-BGT. A better correlation was observed between 3-h knockdown activities (KD50) against German cockroaches (Blattella germanica) and IC50 values obtained from [3H]EPI assays than between 24-h lethal activities (LD50) and IC50 values. While the results indicate that dinotefuran and its analogues interact with the ACh-binding site in cockroach nAChRs, it remains to be elucidated why they displayed lower potencies than those expected based on their insecticidal activities. PMID- 11852646 TI - Quantification of plasma and egg 4,4'-dinitrocarbanilide (DNC) residues for the efficient development of a nicarbazin-based contraceptive for pest waterfowl. AB - Urbanization and associated landscaping has increased the abundance of year-round habitat for waterfowl, resulting in vegetation damage, loss of recreational activities, air transportation mishaps and health hazards. As part of a research program to develop socially acceptable techniques for management of pest bird populations, we are evaluating nicarbazin as a contraceptive in pest and surrogate avian species. As reproductive studies with Canada Geese (Branta canadensis) are tedious due to the difficulty of conducting controlled field studies and/or breeding geese in captivity, we evaluated the effects of oral nicarbazin administration on the production and hatchability of chicken eggs. Blood plasma and egg DNC concentrations were correlated to contraceptive efficacy. Subsequent studies are being conducted with geese to determine the diet nicarbazin concentration required to produce the desired blood and plasma DNC concentrations. This approach permits the expeditious evaluation of formulations and dosing regimes by simply monitoring blood DNC concentrations in target species. PMID- 11852647 TI - Cotton whitefly (Bemisia tabaci) resistance to organophosphate and pyrethroid insecticides in Pakistan. AB - Resistance to three organophosphate and four pyrethroid insecticides was monitored from 1992 to 2000 in field populations of adult whiteflies, Bemisia tabaci, from Pakistan using a leaf-dip method. There was generally a very high resistance to dimethoate and deltamethrin, and a moderate resistance to monocrotophos during 1992 to 1996. From 1997 to 2000, resistance to these insecticides dropped to low levels because of less reliance on them for whitefly control, and introduction of new chemistries with novel modes of action that had no cross-resistance to conventional insecticides. Concurrently, whitefly resistance to acephate, fenpropathrin, lambda-cyhalothrin and bifenthrin mostly remained low. An insecticide resistance management strategy is recommended that particularly emphasizes the rotation of still-effective insecticides from different chemical classes along with the use of novel chemicals and other tactics of integrated pest management. PMID- 11852648 TI - Linkage of genes for sodium channel and cytochrome P450 (CYP6B10) in Heliothis virescens. AB - Genetic linkage of hscp (heliothis sodium channel protein) and CYP6B10 was discovered in Heliothis virescens. The hscp gene encodes the sodium channel target of pyrethroid insecticides and cytochrome P450 genes encode important enzymes involved in detoxication of various pesticides. Previously, two mechanisms, nerve insensitivity due to sodium channel and synergism by propynyl aryl ethers, were observed in pyrethroid-resistant H virescens and were not separated by repeated back-crossing. We hypothesized genetic linkage of target site insensitivity and monooxygenase-mediated detoxication. Single nucleotide polymorphisms were discovered in IIS6 of hscp; Hpy of hscp and CYP6B10. Segregation of these and other markers was tested in backcrosses. We observed cosegregation of hscp to CYP6B10, but both genes assorted independently of y, ye and sex. Genes y and ye assorted independently of each other. This was the first observation of linkage between genes controlling detoxication and sodium ion channel insensitivity in a species known to express high levels of pyrethroid resistance. Linkage was not likely because this species has 31 chromosomes; therefore, we will investigate the possibility of a resistance cassette. We expect similar linkage in other noctuid pests. PMID- 11852649 TI - Acute myocardial infarction at a university hospital: effect of race on short term mortality. AB - Several studies have shown that African Americans who have an acute myocardial infarction (AMI) have a higher mortality rate and receive less aggressive cardiac intervention compared with whites. This observational study reports on the in hospital mortality rate for AMI by race at a university tertiary referral hospital. Racial variation in clinical outcomes and treatment in the area of thrombolysis and acute revascularization was assessed. Data were retrieved from the National Registry on Myocardial Infarction (NRMI) for all 521 patients with AMI admitted to the critical care unit at the University of North Carolina between January 1991 and December 1994. Information collected included age, gender, race, cardiac catheterization results, thrombolytic therapy, coronary bypass surgery, mortality, and arrhythmia. African Americans had a lower in hospital mortality rate compared with whites (2% versus 8% P < or = 0.03) and were also younger (61 +/- 13 SD versus 64 +/- 12 P = 0.02). Hypertension, diabetes mellitus, or prior myocardial infarction were similar in both groups. Of the 323 patients who received a cardiac catheterization, the extent of coronary disease and left ventricular ejection fraction was similar in both races. Finally, the use of thrombolysis, PTCA, or CABG was not influenced by race. In conclusion, the in-hospital mortality for African Americans at this university tertiary referral center was lower than for whites. This occurred despite a similar incidence in cardiac risk factors and similarly aggressive acute cardiac interventions in both white and African American patients. While African Americans experienced lower in-hospital mortality, this study does not address the pre-hospital and post-hospital risk. It does suggest that African Americans with AMI, who are comparably matched to whites for risk and receive similar cardiac interventions, may have a favorable in-hospital mortality. PMID- 11852650 TI - Cervical cancer--the impact of clinical presentation, health and race on survival. AB - This study evaluated the association of race, comorbid illness, insurance status, and other prognostic factors on treatment and survival of patients with invasive carcinoma of the cervix, using as its methodology a retrospective study of patients with invasive carcinoma of the cervix treated from 1991 to 1998 at the University of Maryland. Of 161 such patients, 153 met the study criteria. Of these 153 women, 83 (54%) were African Americans and 70 (46%) were white. The mean (+/- SE) age of African American women was 52 +/- 1.8 years and that of white women was 50 +/- 1.8 years. African American women were more likely to present with stages II-IV disease (P = 0.01) and, as a consequence, underwent radiation therapy rather than surgery (P = 0.04). The survival of African American women with stage I-II disease was significantly lower than for white women. In the final regression model, stage III-IV disease (hazard ratio 3.2, 95% (CI 1.7, 6.1) African American race, (hazard ratio 1.9; 95% CI 1.0, 3.3) and comorbid illnesses (hazard ratio 2.3; 95% CI 1.3-4.0) were associated with poor survival. Adverse outcomes of African American women with cervical cancer persist after adjustment for stage of disease and other factors and are particularly apparent in stage I-II disease. The role of comorbid illness is deserving of further study. PMID- 11852651 TI - Suburban versus urban: does it matter where the residency interview begins? AB - Geographic location is an important factor in students' ranking of residency programs. Our program's inner city location has a negative impact on our recruitment efforts. In an attempt to assess the impact of geography, we started half of the residency interview days at our suburban community medical center and then measured the effect on the applicant's perception of the program. During the 1998-1999 residency interview season we alternated the site at which the day began. Students were randomly assigned based on interview date requests to starting the day at the urban hospital or at the community hospital. At the conclusion of the day the students completed a questionnaire regarding various components of the interview day and how their perception of the program was influenced by the experience. Of 206 students asked to complete the questionnaire, 188 (91%) completed the survey. The degree to which the students' perception of the program was affected was remarkably similar regardless of where the interview day began; however, significant differences were found between the Chairman's Talk, the Teaching Session, and talking with current residents when compared by univariate analysis. We concluded that students' perception of the program at the conclusion of the interview day was similar regardless of whether the interview day began at an urban or suburban medical center. PMID- 11852653 TI - [The physiology of pain]. PMID- 11852652 TI - [History of pain and pain treatment]. PMID- 11852654 TI - [The key-lock model of molecular biological characterization. Opiate receptors]. PMID- 11852655 TI - [Two complex action principles in a single structure. Mixed opioid agonists antagonists and partial agonists]. PMID- 11852656 TI - [Old and new mu-opioids. Mu-selective opioids with a morphinan structure]. PMID- 11852657 TI - [Opioid peptides. Ironman, mother's milk and toad's skin]. PMID- 11852658 TI - [Agonist or antagonist? Structure-activity relationships of opioids]. PMID- 11852659 TI - [The clinical importance of opioids. Postoperative pain therapy]. PMID- 11852660 TI - [Situational administration of analgesics. Modern dosage forms for opioids]. PMID- 11852661 TI - [Meaningful patient-oriented service! Pharmacy information for pain patients]. PMID- 11852662 TI - [Blessing or problem child? Substitution therapy for opiate-addicted patients]. PMID- 11852663 TI - [Why gamble with pain and analgesia? New approaches in central analgesia]. PMID- 11852664 TI - The future role of historically black colleges of medicine. PMID- 11852665 TI - Chemical terrorism: a primer for 2002. AB - Agents of chemical warfare have inflicted mass casualties on military and civilian personnel. Although not yet deployed on populations in the United States, stockpiles exist in countries that view the United States with enmity. There are four groups of such agents. The inhalants, chlorine and phosgene, were the earliest such weapons. Cyanide gases, because of their volatility, exert their lethal consequences in enclosed spaces. The vesicants and nerve agents, currently the most successful agents of chemical mass destruction, are the major focus of this overview. Mustard, the prototype of the vesicants, and its sulfur and nitrogen analogues are aerosolized liquids that linger on victims, incapacitating through their effects on the skin, eye, and respiratory tract. Mortality is low, but morbidity is substantial and mass casualties strain health care resources. Nerve agents, by contrast, are designed to kill. All, including Tabun, Sarin, Soman, and VX, are organophosphates that inactivate acetylcholinesterase. Widespread manifestations of peripheral and central neuronal dysfunction can be fatal without timely administration of antidotes. Decontamination, axiomatic in the management of agents of chemical warfare, is outlined, and therapeutic strategies are summarized in the light of current understanding. PMID- 11852666 TI - Vaccines for adults in an age of terrorism. AB - Vaccines are an effective, safe, and relatively inexpensive means of preventing infection; thus, they are important tools for fighting biological terrorism. Two diseases, anthrax and smallpox, for which vaccines are not available to the general public are discussed. Three other vaccines--tetanus toxoid, influenza vaccine, and hepatitis B vaccine--generally recommended for adults, may be in short supply as a result of recent acts of terrorism. PMID- 11852667 TI - Inhibition of platelet aggregation and expression of alpha granule membrane protein 140 and thromboxane B2 with pravastatin therapy for hypercholesterolemia. AB - The drugs in the group of the "statins" lower blood lipids, especially cholesterol, thereby reducing a risk factor for, and diminishing the incidence of, clinically important cerebrocardiovascular events. Cardiovascular events and stroke are significant causes of morbidity and mortality in China and the United States. Statins reduce platelet-mediated thrombus formation and atherosclerotic progression through mechanisms not completely elucidated. While important, the lipid-lowering action of statins does not completely explain their multifaceted benefits. Nonlipid related mechanisms are essential to such effects. The authors explore these nonlipid related mechanisms of action of pravastatin that may translate into clinically relevant benefits. This study was conducted in Guangzhou, China. Twenty-one hypercholesterolemic patients were treated with pravastatin--10-20 mg/day for 12 weeks. Blood for tests was obtained at baseline and after 8 and 12 weeks of pravastatin therapy. After 8- and 12-weeks of therapy, significant decreases were observed in the following: (1) total blood cholesterol and low density lipoprotein-C (P < 0.01), (2) ADP-induced maximum platelet aggregation (P < 0.01), (3) TXB2 or thromboxane B2 in platelets (P < 0.01), and (4) expression of GMP-140 or granule membrane protein-140 (P < 0.01). The therapeutic effects of the drug did not vary significantly with length of therapy. Pravastatin induces inhibition of platelet aggregation and expression of TXB2 and GMP-140, the likely causes of thrombus formation, atherosclerotic progression, and subsequently cardiovascular events. These potential beneficial events occur within 8 weeks of pravastatin therapy. PMID- 11852668 TI - Statins, platelet aggregation and coronary heart disease. AB - It is becoming increasingly recognized that the beneficial effects of 3-hydroxy-3 methylglutaryl coenzyme A (HMG-Co-A) reductase inhibitors (statins) on reducing clinically important cardiovascular events (myocardial infarction and stroke) are not only attributable to their hypocholesterolemic effect but also to non-lipid mechanisms of action. The nonlipid factors may include the stabilization of arterial plaques, endothelial normalization, anti-inflammatory effects and inhibition of platelet thrombus formation. The inhibition of platelet thrombus formation has not been adequately studied in man and the results are often contradictory. It is the objective of this review to discuss the effects of statins on platelet aggregation in the context of relatively new and specific techniques for the measurement of platelet function as reported by Ma and coinvestigators in this issue of JAAMP. Ma et al. examined the effect of pravastatin given for 8 to 12 weeks on platelet function and low density lipoprotein-cholesterol (LDL-C) in 21 Chinese patients with primary hypercholesterolemia. Platelet function was evaluated by adenosine diphosphate (ADP)-induced aggregation, thromboxane B2 (TXB2) and the expression of alpha granule membrane protein-140 (GMP-140). GMP-140 is considered one of the most sensitive indicators of the state of platelet function. As expected, pravastatin treatment significantly reduced LDL-C and inhibited ADP-induced platelet aggregation, TXB2 synthesis and the expression of GMP-140--all such parameters can lead to thrombus formation and subsequent cardiovascular events. Using the test methods of Ma et al., additional dose-response studies with several statins and standard antiplatelet drugs are needed to confirm their effects on platelet aggregation, if any. Furthermore, we need to determine whether the antiplatelet effect of the statins, if present, is independent of their hypocholesterolemic action. The additional studies could provide important clues toward the development of new and specific antithrombotic drugs. PMID- 11852669 TI - Bioterrorism: a primer for 2002. AB - The often anticipated, sometimes dismissed, threat of biological terrorism became a reality in the United States in the fall of 2001 with cases of cutaneous and inhalation anthrax. As the public health sector is rapidly focusing on enhancing awareness and preparedness, this overview is intended as a primer for clinicians. The more probable events, their most prominent characteristics, and currently available management strategies are summarized. The biowarfare characteristics of plague, tularemia and botulism are summarized. The more problematic anthrax and smallpox are presented in greater detail. Epidemiological clues to a biological attack are outlined with reference to the recent literature. PMID- 11852670 TI - Quality alternatives in medicine. PMID- 11852671 TI - CAM providers' messages to conventional medicine: a qualitative study. AB - Between 30 and 50% of patients use complementary and alternative medicine (CAM). There is little research on the interaction between CAM and conventional providers. We investigated what messages CAM practitioners would convey to conventional medicine (CM). Thirty-four CAM practitioners participated in audiotaped interviews. A coherent message was constructed from the identified themes. CAM practitioners see CAM supporting CM rather than replacing it. Blending of CAM with CM benefits patients and CM providers. CAM reintroduces the concept of healing that technology and time pressures have reduced. The basis of healing is connection, being present in the moment, and seeing patients as whole human beings. Research validating CAM effectiveness will foster integration, as will inclusion of CAM theory and practice in the medical curricula. The messages from CAM practitioners to CM coincide with current views of integrative medicine. Collaboration in research, education, and practice can foster a high-quality health care system. PMID- 11852672 TI - Credentialing of practitioners of botanical medicine. AB - This article discusses how practitioners, regardless of other professional licenses they may hold, could be credentialed in botanical medicine. The article reviews the field of clinical botanical medicine and the history and modern status of botanical medicine, as well as organizations currently involved in botanical medicine credentialing. Many different types of professionals prescribe botanical medicines, and the potential for collaboration among them is great. The current trend treats botanical medicine as a narrow subdivision of allopathic medicine and does not acknowledge the breadth, depth, and diversity of botanical medicine and ultimately will not provide maximum benefits for patients. An alternative approach that instead credentials practitioners skilled in the use of a wide variety of botanical medicines in a responsible, scientific fashion is presented. PMID- 11852673 TI - The use of complementary and alternative therapies among middle-aged and older cardiac patients. AB - This study investigated the prevalence, pattern, perceived benefits, and predictors of the use of complementary and alternative therapies (CAM) among middle-aged and older patients prior to cardiac surgery. Comprehensive information of 246 patients who came for preoperative medical assessment was obtained through face-to-face interviews. CAM utility of 225 was obtained by telephone the day before surgery. A computerized STS Adult Cardiac Database of Cardiac Surgery provided patients' medical information. One hundred eighty-two patients (80.9%) confirmed CAM use, including relaxation techniques, lifestyle diet modification, spiritual healing, megavitamin therapy, massage, herbs or folk remedies, and imagery. Education, functional status, the number of non-cardiac chronic conditions, status as former cigarette smoker, public religiosity, and congestive heart failure predicted greater CAM utility, whereas cerebrovascular diseases and arrhythmia tended to predict less use. Findings suggest that there is considerable CAM use in this cardiac sample. Physicians and other health providers should investigate patients' CAM utility and promote collaborative medical care. PMID- 11852674 TI - Commentary: alternative medicine: is it a reflection of the continued emergence of the biopsychosocial paradigm? AB - The possibility is presented that the current collaborative versus conventional medical debate may be a consumer-driven continuation of the biomedical versus biopsychosocial controversy first proposed by George Engel 25 years ago. If true, we may all be actors in a genuine Kuhnian paradigmatic shift destined to occur whether we appreciate our roles therein or approve of the ultimate outcome. At the very least, however, it is important to recognize the evidence that at least some patients suffering from chronic conditions may be choosing collaborative therapies for different reasons than for those for which they seek conventional medical care. For these patients, at least, it is important that the quality of the collaborative care they receive be judged on outcomes capable of reflecting these alternative objectives. Finally, whether we are in the midst of a paradigmatic shift in medicine or not, it is essential that we all, as providers of clinical care or as investigators of clinical efficacy, adopt a collaborative role to ensure the provision of both good medicine and good science. PMID- 11852675 TI - Academic physicians and complementary and alternative medicine: an institutional survey. AB - In order to assess attitudes, awareness, and behavior related to complementary and alternative medicine (CAM) among academic physicians, a questionnaire study was conducted with faculty in one of the largest teaching hospitals in the United States. The survey assessed attitudes toward and awareness of 11 CAM therapies in regard to patient usage and usefulness. The majority of respondents believed that only 20% or less of their patients used CAM. Respondents varied widely on level of knowledge/awareness of specific CAM therapies. Those who were more knowledgeable were more likely to perceive CAM as useful and to have recommended more therapies. The modalities respondents noted as knowing the least about were deemed the least useful. Limited knowledge of most CAM therapies and limited awareness of patient CAM use points to the need for continuing education efforts for physicians, focusing on CAM. PMID- 11852676 TI - Commentary: patient-centered care, the sine qua non of collaborative medicine. AB - Collaborative medicine requires practitioners and disciplines with varied training and philosophies to work together toward a common clinical goal. Distinct differences between the various practitioners and disciplines can serve as both opportunities for synergy and barriers to collaboration. There must be a clear and compelling reason that motivates different practitioners and disciplines to do the hard work needed to overcome barriers to collaboration. Patient-centered care is the compelling reason, the defining characteristic that must underlie all collaborative medicine efforts. The practical application of this principle is illustrated by the experience of a large health care system (Catholic Health Initiatives). From a complexity theory standpoint, patient centered care and collaboration can be thought of as simple rules that guide desirable behaviors in a complex system. The application of these two simple rules is as relevant in the microcosm of collaborative medicine as it is in the larger macrocosm of health care. PMID- 11852677 TI - [Brain emboli in the lungs of cattle]. AB - There is no information whether the BSE agent is introduced into the human food chain through contamination of the lungs of cattle with central nervous system tissue (CNS). Studies in the United Kingdom and in the USA showed that CNS tissue could contaminate the lungs after using pneumatic powered air injection stunners (e.g. "The Knocker") or after pithing. Thus, pithing was forbidden in the European Union since January 2001. In German abattoirs conventional cartridge fired stunners (e.g. model by Schermer) are usually applied. Pithing was used up to December 2000 in approx. 75% of the German abattoirs. In the present study 323 lungs of cattle were analysed for CNS. The lungs were derived from cattle exclusive stunned by use of the knocker from Schermer. 60% of the lungs contained emboli which were tested with immuno chemistry as well as immuno histochemistry to detect CNS. Two of 108 pooled samples showed a faint immuno reaction in the anti-NSE and anti-GFAP immunoblot. Further two particles showed a faint reaction for NSE and GFAP in immuno histochemistry, thus suggesting the presence of CNS. Even though CNS tissue could not be shown in the histological investigation, we used our findings to estimate the worst case scenario for human BSE exposure risk (HER) by lung contaminated by CNS emboli. The content of CNS in the samples was estimated to be about 0.11% when the respective immuno reactions were calibrated against standards containing known brain concentrations. Under the assumption that only one lung in the pooled samples was contaminated with BSE-infected central nervous tissue, the HER was calculated to reach a maximum of 2.2 x 10(-5) CoID50/consumer after consumption of a sausage with a portion of 10% lung. The results of our study suggest that the contamination of the lung with CNS after using a conventional cartridge-fired stunner cannot be excluded, however, the incidence appears to be very low. In addition, presumed CNS emboli, if at all, are microscopically small. Furthermore the incidence of BSE in Germany is very low and lungs of cattle are usually not consumed. Thus we can judge the potential for human oral exposure after consumption of lungs of cattle which were stunned in Germany to be extremely low. A final assessment, however, is impossible as there is no knowledge about the minimum infectious dose for humans. PMID- 11852678 TI - [The redox status of the blood plasma in cattle from ecological farming--1. Influence of genotype and age on the antioxidant defence capacity in the blood plasma]. AB - The present study examined the effects of race and age on the redox potential behavior of blood plasma samples. The oxidative stability was studied in the reaction with p-benzoquinone. The investigated animals represented three different cattle races (Limousin, Angus and Hereford), grazed on pasture. The blood samples from clinical healthy heifers (44) and calves (45) were investigated. The following parameters were determined: the start level of redox potential and the level after adding 10, 20, and 30 mg p-benzoquinone to the samples. The analyses of the redox potentials showed the advantage of oxidative stability of heifers compared with calves. The results showed a significant effect of race on the redox potential behavior of blood plasma. PMID- 11852679 TI - [The effectiveness of rare earth elements as a possible alternative growth promoter for pigs and poultry]. AB - Mixtures of Rare Earth Elements (REE) have been used as animal growth-promoters on a large scale in China during the last 20 years. Numerous studies carried out in China claim it produces quite sensational growth-promoting effects in all categories of farm animals. To explore the question of whether REE's might prove suitable as a growth-promoter under western keeping conditions, feeding experiments were performed on pigs and poultry. The animals received a typical diet, supplemented with REE salts in concentrations between 75 and 300 mg/kg feed. Weight-gain, feed-intake, feed-conversion and (where applicable) laying parameters were observed. It was shown that in pigs receiving feed supplemented with REEs, an increase in daily weight gain of up to 19% and an improvement in feed-conversion of up to 11% can be achieved, whereas, for poultry, no positive effects on growth or productivity of the animals could be observed. Testing of important organs via Neutron Activating Analysis (NAA) showed a minute accumulation of REE, principally in liver and bones. Analysis of the poultry gut flora, using selective media, showed that the main microorganism populations of the alimentary canal were unaffected by feed-supplementation with REE. PMID- 11852680 TI - [Quantitative determination of lipases and their heat stability in food of animal origin]. AB - Quality deterioration and food spoilage in fat-containing food is mostly caused by bacterial and endogenous lipases. Quantitative data on the concentration of lipases for a possibly better evaluation of shelf life do not exist. Own investigations for detecting lipases and evaluating their heat stability in selected food of animal origin (meat-products, fish-products, porcine liver) were carried out using the Reflektoquant Lipasetest (Merck). Extreme high values could be found in porcine liver and in muscles of fat fish such as herring. Pork contained 46 micrograms/kg, beef 86 micrograms/kg and poultry a 3-fold higher value. Significant differences could be observed in parts within poultry (breast 139 micrograms/kg, leg 259 micrograms/kg). Heated sausage and hot smoked fish were largely lipase-free, raw sausages contained up to 283 micrograms/kg lipases. Fish- and meat-lipases were not as heat stable as investigated bacterial lipases which makes them distinguishable. PMID- 11852681 TI - [Air microorganisms in animal housing--4. Airborne gram-negative bacteria and airborne endotoxin in pig houses]. AB - The species composition of airborne gram-negative bacteria and the relationship between inhalable endotoxin, inhalable dust and airborne bacteria were studied in 4 pig houses. The airborne gram-negative bacterial flora was dominated by Enterobacteriaceae. Within the Enterobacteriaceae the species E. coli and Enterobacter agglomerans were predominant. Significant correlation were found between the concentration of inhalable dust and inhalable endotoxin as well as between the concentration of airborne gram-negative bacteria and inhalable endotoxin. However these correlation were not very strong. With respect to the characterization of the potential hazard of organic dust exposure, measurements of the concentration of airborne dust or airborne bacteria should not be used for the estimation of the concentration of airborne endotoxin. PMID- 11852682 TI - Effect of highly fermentable dietary fiber on the development of swine dysentery and on pig performance in a "Pure--Culture Challenge Model". AB - This study tried to evaluate the effect of highly fermentable fiber on the incidence and severity of swine dysentery (SD) after experimental oral infection with pure cultures of Brachyspira (B.) hyodysenteriae. Forty eight growing pigs were allocated to two groups and treated until slaughter as follows: Group 1 (n = 24): infected with B. hyodysenteriae and fed with a food containing 9.6% highly fermentable neutral detergent fiber. Group 2 (n = 24): infected with B. hyodysenteriae and fed with a food containing 6.1% low fermentable neutral detergent fiber. Pigs of each group were intragastrically inoculated on each of three consecutive days with pure culture of 1.8 x 10(10) B. hyodysenteriae. All pigs were monitored daily until slaughter. Faecal shedding of B. hyodysenteriae by polymerase chain reaction, antibody response by IFA, clinical signs, growth performance and extents of gross and microscopical lesions specific for swine dysentery were determined. Faecal shedding of B. hyodysenteriae and antibodies specific for B. hyodysenteriae were detected at day 30 post infectionem. Significant (p < 0.05) milder clinical signs typical for swine dysentery were detected in group 1, fed with 9.6% high fermentable fiber compared to group two fed with a food containing 6.1% low fermentable neutral detergent fiber. Daily weight gain differed significantly (p < 0.05) between the groups (group one 780 g vs. group two 760 g). Food conversion efficiency showed in group one a significant (p < 0.05) better (3.28) result than in group two (3.38). Feed consumption presented significantly (p < 0.001) better results in group one compared to group two (2.38 kg vs. 2.25 kg). From our experimental findings we conclude that in production units suffering of swine dysentery high levels of highly fermentable fiber in diet may increase health and performance. PMID- 11852683 TI - [Occurrence of parasites in indigenous birds of prey and owls]. AB - In the present paper a general overview on parasites in birds of prey and owls is given. This part is followed by a study investigating the prevalences and species of parasites in free-ranging birds of prey and owls in Berlin and Brandenburg State, Germany. Over a one year period, 84 birds of prey and owls of the following species were examined for the presence of endo- and ectoparasites: Common Buzzard (Buteo buteo) (n = 32), Kestrel (Falco tinnunculus) (n = 20), Sparrowhawk (Accipiter nisus) (n = 9), Goshawk (Accipiter gentilis) (n = 8), Black Kite (Milvus migrans) (n = 4), Peregrine Falcon (Falco peregrinus) (n = 3), Marsh Harrier (Circus aeruginosus) (n = 1), White-tailed-Sea Eagle (Haliaeetus albicilla) (n = 1), Tawny Owl (Strix aluco) (n = 4), Long-eared Owl (Asio otus) (n = 1) and Barn Owl (Tyto alba) (n = 1). In 97.6% of the cases, ectoparasites (feather mites and hippoboscid flies) were found. Especially eyasses (93.3%) were positive for hippoboscid flies. Trichomonas was detected in 28.6% of all birds of prey and owls examined. A prevalence of 100% was established in the Sparrow Hawks as well as Peregrine Falcons. Leucozytozoon sp. and Hemoproteus sp. as blood parasites were found in 26.9% of the birds in total. Common Buzzards showed the highest prevalence (44.8%). 58.3% of birds examined were positive for endoparasites. Flukes were found in 16.7%, tapeworms in 14.3%, round-worms in 48.8% and acanthocephales in 2.4% of the cases. Interestingly, Tylodelphis clavata (in a Common Buzzard) and Hovorkonema variegatum (in a Goshawk) were found for the first time in raptors. The results of this study underline the importance of a parasitological examination in the process of raptor rehabilitation. PMID- 11852685 TI - [Pharmacokinetics of cephalexin from two oral formulations in dogs]. AB - Six beagle dogs were treated with cephalexin-monohydrate from 2 oral formulations (Rilexine tablets and Cefaseptin dragees, respectively) in a dosage of 25 mg/kg and plasma concentrations of cephalexin were measured over 8 hours. After solid phase extraction of the samples, cephalexin was determined by high pressure liquid chromatography with UV detection. After administration, Cephalexin was absorbed rapidly and mean maximum plasma concentrations of 30.9 and 27.9 micrograms/ml, respectively, were acquired after approximately 1.6 hours. Minimal inhibitory concentrations of < or = 6.25 micrograms/ml for in vitro sensitive bacteria were maintained for about 5 hours. Cephalexin from the tested preparations reached a mean area under the plasma concentration-time curve of 115.3 and 102.4 micrograms.h/ml, respectively. The plasma concentration decreased rapidly with a mean half life period of 1.4 hours in average. The other calculated pharmacokinetic parameters were also in the area of the data for dogs stated in the literature. There was no clear difference in the pharmacokinetics of both products, especially the bioavailability. Furthermore, both formulations were well tolerated clinically. PMID- 11852684 TI - [10 years experience with denervation of the hip joint capsule for treatment of canine hip joint dysplasia and arthrosis]. AB - Denervation of the canine hip joint capsule is described as a surgical therapy method in the treatment of canine hip joint dysplasia and arthrosis. The goal of this operation is a removal of the pain immediately and a reactivation of the dynamic active component of the hip joint in moving the body forward. Simple removal of the periosteum of the craniolateral acetabulum edge destroys the rami articulares of the cranial glutaeal nerve and the rami articulares dorsalis of the sciatic nerve. Within 10 years now we performed the denervation of the canine hip joint capsule. Evaluation of the post operative clinical course in 269 cases revealed an impressive improvement of lameness due to pain relief in almost 92% of the cases. PMID- 11852686 TI - Effect of nisin and monensin on rumen fermentation in the artificial rumen. AB - The objective of this study was to investigate the effect of nisin and monensin on rumen fermentation of diets containing hay and barley (80:20%) in artificial rumen (Rusitec system). The Rusitec system consisted of four fermentation vessels (V1, V2, V3, V4): V1 was without additives (control), V2 received daily 2 mg of nisin, V3 involved 5 mg of monensin and V4 combination of 2 mg of nisin with 5 mg of monensin. After an adaptation period (7 days), the fermentation parameters were determined for six consecutive days. Compared to control diet, the addition of nisin resulted in an increase (P < 0.05) of hemicellulose degradation, acetate, propionate (mmol.day-1) production and energetic efficiency of VFA (E), decrease of butyrate production. Nisin had no effect on dry matter (DM), organic matter (OM), cellulose and detergent fiber degradation, production of total gas, methane and efficiency of microbial synthesis. The addition of monensin resulted in an decrease of DM, OM (P < 0.05), cellulose, hemicellulose, detergent fiber degradation (P < 0.001), total gas, methane and ammonia nitrogen (NH3-N) production. Monensin also significantly decreased acetate, butyrate, L-lactate (mmol.day-1) production and it increased propionate production (P > 0.001) and efficiency of microbial synthesis. The combined effect of nisin and monensin in V4 was similar to the effect of monensin in V3 compared to control. Then, the effect of additive monensin was dominant over nisin. In conclusion, our results indicate that nisin was less effective than monensin on some fermentation parameters (important for the improvement of the efficiency of utilization of the diet by ruminants) in artificial rumen. PMID- 11852687 TI - Participatory action research: integrating community occupational therapy practice and research. AB - Participatory action research (PAR) is a form of collaborative research particularly concerned with power inequities of marginalized communities. With this approach, occupational therapists can work with clients and communities to address issues of access, inclusion, equity and collaboration in practice and research. This paper begins with a summary of key concepts and controversies related to participatory action research. Two examples from occupational therapy experience are used to demonstrate the initial steps and key principles of PAR projects and to highlight the conceptual links between PAR and community-based, client-centred occupational therapy. One project involved a group of mental health consumers, the other a community group of older adults. Several challenges and potential outcomes in PAR projects are highlighted in this examination of the complexity of PAR processes. The paper concludes by identifying possibilities and obstacles to the further use of PAR in occupational therapy. PMID- 11852688 TI - [Theories emerging from daily clinical practice of community occupational therapists]. AB - Occupational therapists working in the community have adapted their knowledge to the distinct characteristics and expectations of the various settings. To do so, they have had to reflect on the professional nature of their interventions, while intervening. This simultaneous process has yielded specific knowledge that is solidly anchored in current clinical reality. A survey on occupational therapy in the Centres locaux de services communautaires in Quebec (CLSC) highlighted three theories developed by occupational therapists in their daily clinical practice. The first relates mostly to referral processing while the other two address the occupational therapist's role within the treatment team. This paper briefly presents the survey's results, then focuses mainly on the three emergent theories and underlines some elements congruent with the reference theories in occupational therapy. The author then proposes avenues for reflection to help foster balance between the constraints inherent to the community health care system and occupational therapy professional values. PMID- 11852689 TI - Clinical utility of the Canadian Occupational Performance Measure--Swedish version. AB - The Canadian Occupational Performance Measure (COPM) is an individualised outcome measure intended to detect change in a client's perception of occupational performance over time. The aim of this study was to test the clinical utility of the Swedish version of the COPM. Data was collected from 27 occupational therapists in six focus groups. Emerging themes included goal-setting, preparations, limitations, interactions with clients, and impact on practice. The results indicated that the COPM is helpful in the goal-setting process and in planning treatment interventions. Therapists need knowledge about the theoretical foundation of the instrument and a personal interview technique. Problems were found using the instrument with clients who had poor insight or in acute settings. The COPM facilitated feedback on improvement over time. In summary, the COPM ensures a client-centred approach, facilitates communication within the rehabilitation team, and encourages therapists in their professional role. PMID- 11852690 TI - Development of a questionnaire to examine confidence of occupational therapy students during fieldwork experiences. AB - This paper describes the development of an instrument designed to examine the level of confidence of occupational therapy students during their fieldwork experiences. Confidence in this context refers to Bandura's concept of self efficacy in Social Cognitive Theory. As this subject has received little formal study, no appropriate tool was found to measure confidence in students. The purpose of this study was to design a measure to provide students with an opportunity to self-assess their levels of confidence both within a placement and throughout their entire fieldwork experiences. The instrument was constructed in three phases: I--Instrument Development; II--Instrument Testing; 111--Instrument Implementation. In Phase III, the instrument was distributed to all students entering the first year of occupational therapy studies at the University of Alberta. Data was collected from these students during their four fieldwork placements to address the questions of whether confidence increases during a fieldwork placement regardless of placement level, whether confidence increases by placement level, and whether student or placement characteristics affect levels of confidence. Results of the study support the hypotheses that confidence levels increase both during a fieldwork placement and with successive placements. Reliability and validity analyses indicated that the measure may be a useful tool to assess confidence in students during fieldwork placements. PMID- 11852691 TI - Spirituality unplugged: a review of commonalities and contentions, and a resolution. AB - Spirituality is an important and essential component of occupational therapy, but recent publications in occupational therapy literature also raise questions about the adequacy of the Canadian Association of Occupational Therapists' definition of spirituality and the relationship of spirituality to occupational performance. Re-examination of spirituality and occupation is needed to better understand the role of occupational therapists with respect to spirituality. In this paper, the authors examine the common themes that are inherent in definitions of spirituality from diverse professional perspectives. The commonalities and contentions inherent in these definitions are then contrasted with the perspectives of occupational therapists. This discussion is followed by a challenge for re-examination of spirituality in the Canadian Model of Occupational Performance, and an argument that occupational identity rather than spirituality should have a central position. The implications of spirituality and occupational identity for evidence-based occupational therapy are considered. PMID- 11852692 TI - Stabilization of pharmaceuticals to oxidative degradation. AB - A guide for stabilization of pharmaceuticals to oxidation is presented. Literature is presented with an attempt to be a ready source for data and recommendations for formulators. Liquid and solid dosage forms are discussed with options including formulation changes, additives, and packaging documented. In particular, selection of and methods for use of antioxidants are discussed including recommended levels. PMID- 11852693 TI - Mechanism for further enhancement in drug dissolution from solid-dispersion granules upon storage. AB - The present study was performed to investigate the further increase in drug dissolution on storage of ternary solid-dispersion granules containing poorly water-soluble drugs. Ternary solid-dispersion granules of the drug, a dispersion carrier, and a surface adsorbent were prepared using hot-melt granulation. Two proton-donating drugs, BAY 12-9566, naproxen, and a nonproton-donating drug, progesterone, were studied. Gelucire 50/13 and polyethylene glycol 8000 were evaluated as solid-dispersion carriers with low melting point. Neusilin US2 (magnesium aluminosilicate), a proton acceptor, was used as the surface adsorbent. The proposed mechanism for further increase in drug dissolution (BAY 12-9566 and naproxen) on storage at 40 degrees C/75% RH (relative humidity) is based on hydrogen bonding between the proton-donating drugs and the surface adsorbent, Neusilin US2 (proton acceptor). We propose that there is enough mobility in the solid-dispersion granules at elevated temperatures of storage to allow an increase in the ratio of the hydrogen bonded drug to the crystalline drug. These changes are mediated through the saturated solid solution state, and manifest themselves as increased drug dissolution upon storage. Fourier transform infrared spectroscopy studies are indicative of an increase in the amount of drugs (BAY 12-9566 and naproxen) hydrogen bonded to Neusilin on storage. A corresponding decrease in the crystallinity of these drugs was measured using x ray powder diffractometry. Granules containing progesterone (a nonproton-donating drug) do not show an increase in the amount of drug hydrogen-bonded to Neusilin upon storage. In contrast to the proton-donating drugs, decreased drug dissolution was found on storage of progesterone-containing granules. PMID- 11852694 TI - Sequential experimentation in product development. I. AB - Investigations proceed as an iterative and recursive learning process, involving a sequence of experiments. Evaluating the results from a prior set of experiments allows the investigator to design and predict the next experimental cycle better. The investigator looks at what has gone before with a view to improve and extend the inquiry. This communication reiterates the value of sequential experimentation in product development. A fluid-bed granulation study is used to illustrate the sequential approach. Acceptable solutions were achieved using technical knowledge, a sequence of factorial designs, and graphical data analysis. PMID- 11852695 TI - A novel technique to quantify film--tablet interfacial thickness. AB - The purpose of the project was to develop an x-ray photoelectron spectroscopic technique to quantify film-tablet interfacial thickness. Tablets containing 99% dibasic calcium phosphate were coated with an acrylic polymer containing chlorine (Eudragit RL 30 D). The film-tablet interface was defined as the region where phosphorous from the tablet and chlorine from the film coating were detected simultaneously. Five-minute intermittent ion-sputtering was used in conjunction with the conventional x-ray photoelectron spectroscopy to achieve depth profiling of the sample. The slow ion-sputtering rate of approximately 56 nm/min was overcome through sample preparation and analysis from the tablet side of the sample. This process eliminated the need to sputter through the entire film coating to reach the area of interest. Initially, only phosphorous from the tablet was detected. As the depth profiling proceeded, the chlorine signal increased and the relative intensity of the phosphorous decreased. By plotting the relative intensities of each element against the ion-sputtering time and using linear regression, the point at which the two lines intersected (tmid) was determined. By doubling tmid and multiplying by the estimated ion-sputtering rate, the thickness of the film-tablet interface was estimated to be approximately 34 microns. Further studies are needed to correlate the film-tablet interfacial thickness with the adhesive properties of coated solids. PMID- 11852696 TI - Sorption of benzoic acid, sorbic acid, benzyl alcohol, and benzalkonium chloride by flexible tubing. AB - Earlier studies using methylparaben, propylparaben, and several types of flexible tubing showed that the parabens are sorbed by a number of commonly used tubing. The sorption increased with increase in time of contact and the tubing surface area. The temperature and pH ranges tested did not affect sorption. Current studies evaluated other commonly used antimicrobial preservatives used in oral, parenteral, opthalmic, nasal, and other pharmaceutical products. These preservatives include benzoic acid, sorbic acid, benzyl alcohol, and benzalkonium chloride. Results show that all preservatives, except benzalkonium chloride, are sorbed by several types of flexible tubing. After 24 hr, some types of tubing sorbed over 40% of benzoic acid, benzyl alcohol, and about 30% of sorbic acid. Significant losses were observed within a few hours. No sorption of benzalkonium chloride occurred in the tubing tested. Three different types of fluoropolymer resin tubing, Teflon (flourinated ethylene propylene (FEP), Teflon perfluoroalkoxy) (PFA) and Teflon polytetrafluoroethylene (PTFE, NXT), fluoropolymer FEP laminated Tygon tubing, and Zelite do not sorb any of the preservatives. Silicone tubing shows the highest sorption of preservatives. After 120 hr, as much as 64% of the preservative is sorbed by some tubing. Detailed studies using silicone tubing show that the sorption increases with increase in surface area at 25 degrees C, and increasing the temperature to 40 degrees C shows little change in sorption. At the concentration range generally used in formulations, the concentration of the preservative has no significant effect on sorption. Refilling the tubing with fresh preservative solution after contact with preservative solution for 168 hr still causes extensive sorption of the preservative. Desorption studies indicate that the sorbed preservatives are desorbed in water to a limited extent over 120 hr. Results are important in situations where short or extended interruptions occur in the processing and filling of products. This can lead to decreased and variable results due to residence of the solution in the tubing. PMID- 11852697 TI - The influence of film coating on pellet properties. AB - There are numerous reasons for which film coatings are applied to pellet formulations; for example, controlled release, taste masking, and improved stability. The aim of this paper was to study the influence of pellet shape on the deposition of film coatings in a fluid-bed process by monitoring the pellet shape as a function of the film thickness formed. Eight pellet batches were used, of which four were spherical visually, and the other batches can be described as ovoids, dumbbells, long dumbbells, and cylinders. The average coat thickness of the pellets assessed by cross-section measurements did not appear to be influenced by the initial shape of the pellets. The fluid-bed process, however, had an impact especially for those pellets that had an aspect ratio greater than 1.5. The change in the pellet shape during film coating could only be monitored effectively employing a three-dimensional shape factor. Significant changes in shape occurred at the beginning of the coating process up to approximately the first 15 min, after which the shape remained constant. PMID- 11852698 TI - The effect of erosion and swelling on the dissolution of theophylline from low and high viscosity sodium alginate matrices. AB - The extent of dissolution of theophylline from sodium alginate matrices has been studied in water and HCl. Surprisingly, it was observed that the dissolution rate was very fast for the low viscosity (LV) material in water and for the high viscosity (HV) in acid, whereas slow release was seen for LV in acid with very slow release for the HV in water. The extent of erosion and swelling of the matrices (with and without drug and with and without stirring) was determined from simple weighing experiments. Based on the extent of erosion and swelling, along with visual observations, it was concluded that the rapid release from LV in water was due to rapid and complete erosion of a weak gel layer. The rapid release of HV in HCl was due to an unexpected lamination of the tablet during swelling, which resulted in exposure of more of the tablet core, and caused increased erosion of the matrix. The slow release seen with the LV in HCl was due to the acid gel being tough and resistant to erosion. The very slow release observed with the HV in water was due to very substantial swelling, with the presence of near zero-order kinetics for this system being due to a balance of swelling and erosion, keeping the diffusion process at a constant rate. Furthermore, an anisotropic swelling behavior is indicated by the preferential expansion in the axial dimension relative to the radial dimension for both LV and HV alginates. PMID- 11852699 TI - Comparative evaluation of co-processed lactose and microcrystalline cellulose with their physical mixtures in the formulation of folic acid tablets. AB - In this study, a new co-processed filler-binder ingredient for direct compression, MicroceLac 100, was compared with three different lactoses mixed with microcrystalline cellulose. The aim was to improve a folic acid tablet formulation. Therefore, the influence of drug addition to these mixtures was studied with regard to flow and binding properties. Another part of the study was focused on the interaction and segregation behavior of the drug. MicroceLac 100 showed superior flow and binding properties. Good adhesion of folic acid to the MicroceLac 100 particles could decrease demixing and segregation. The improved characteristics of co-processed material are attributed to spray drying. Sodium stearyl fumarate was chosen as a lubricant, based on ejection force measurements. The content uniformity of the newly formulated direct compression tablets met the official requirements. PMID- 11852700 TI - Estimation of initial dissolution rate of drug substance by thermal analysis: application for carbamazepine hydrate. AB - We have proposed a theory indicating the correlation between the dissolution rate and the heat of solution of drug substances. The initial dissolution rates of the drug substances containing amorphous were predicted accurately from their heats of solution. In this report, the possibility for the theory to estimate the dissolution rates of hydrate and polymorphs was examined using thermal analysis. The initial dissolution rates of carbamazepine dihydrate and polymorphs (forms I, II, and III) were measured by the rotating disk method. The heats of solution and the heats of fusion of samples were measured by microcalorimetry and Differential Scanning Calorimetry (DSC), respectively. The logarithm of the initial dissolution rate of the sample was correlated linearly with the heat of fusion as well as the heat of solution. The obtained correlation would be applicable for the quality control of the drug substances the contained hydrate and/or polymorphic forms. PMID- 11852702 TI - State of pharmaceutics in 2002. PMID- 11852701 TI - A preliminary investigation of modified alginates as a matrix for gene transfection in a HeLa cell model. AB - Previous reports have demonstrated the effectiveness of chitosan as a transfection agent. These studies have noted the importance of polysaccharide backbone interactions with the cell surface as well as cationic groups in the transfection process. The present study focuses upon the potential utility of another polysaccharide hydrogel, alginic acid, as a transfection agent. Alginic acid was modified by carbodiimide-mediated linkage of several heterocyclic and aromatic amines to the carboxyl group of the alginate, giving the alginate polycationic characteristics through which binding to nucleic acids could be facilitated. The amines used for this modification include diaminoacridine, thionin, basic fuchsin, acridine yellow, and diaminomethyltriazine. Of all the conjugates tested, basic fuchsin-modified alginate produced the greatest increase in the transfection of a plasmid coding for beta-galactosidase into HeLa cells. These studies demonstrate that other polysaccharide hydrogels can be used as transfection agents, and the structural orientation of the cationic spacer arm is crucial for effective transfection. PMID- 11852703 TI - SUBSUNK-Royal Navy Medical support to Russian rescue attempt following sinking of the Kursk. AB - On 12 August 2000 a Russian Oscar class submarine, the KURSK, sank in the Barents Sea. This article describes the medical support provided by the Royal Navy in response to this tragic accident. PMID- 11852704 TI - Chilaiditi syndrome and recurrent colonic volvulus: a case report. AB - The case of a 31 year old male presenting as an emergency with a recurrent colonic volvulus is described. A chest X-ray on admission to hospital showed the presence of hepato-diaphragmatic interposition of the colon, Chilaiditi's Sign, which is known to be a risk factor for colonic volvulus. This is only the fourth reported case of colonic volvulus in association with Chilaiditi's Syndrome and the first with recurrent colonic volvulus. The optimal treatment for recurrent volvulus in patients with risk factors such as Chilaiditi's Syndrome or megacolon is also discussed. PMID- 11852705 TI - Three year review following bone anchor repair of acute biceps tendon insertion rupture. AB - Three patients with acute biceps brachii tendon insertion ruptures were treated (less than 7 days post injury) using bone anchor anatomical repair. Dynamometer assessment of strength in both limbs was performed after completion of rehabilitation and again at 3.3 years after surgery. All patients were male, age 34, 35 and 53 years. Early isokinetic assessment was performed at 6, 7 and 12 months post surgery and medium assessment at 3.3 years post surgery. A full range of movement was achieved at early assessment and maintained into medium term assessment. All patients returned to their full premorbid occupation and sporting activity. Dynamometer strength of a repaired dominant limb equated to two thirds of a normal non dominant limb at early assessment, equal power was found at medium term assessment and a measurable increase in strength in both affected and unaffected arms was seen. PMID- 11852706 TI - Lisfranc injury--surgical fixation facilities an early return to work. AB - BACKGROUND: Lisfranc injury is rare and the diagnosis maybe easily missed. This study reviews the experience of a single centre. METHODS: A prospective review of patients with Lisfranc injuries presenting to a single surgeon with a specialist interest in foot and ankle surgery over a one year period. RESULTS: Five patients were identified--four men and one woman with a median age of 31 years (range 22 50 years). Presentation was a mean of 25 days after injury (range 3-56 days). The left foot was affected in three cases and the right in two. There was joint diastasis in four patients and fracture-dislocation in one. Three patients presented early and were treated by internal fixation and two presented late and were managed conservatively. Mean follow-up was eight months (range 4.5-12 months). Surgery resulted in a return to work by 6 months with no symptoms. The two patients managed conservatively continued to experience pain at 12 months and were unable to return to their original occupations. CONCLUSION: Injury to the Lisfranc joint should be excluded in any foot injury. Early diagnosis and internal fixation appears to result in an earlier return to work when compared to non-operative management. PMID- 11852707 TI - A visit to Brooke Army Medical Center, San Antonio, Texas and hospital Escuela, Tegucigalpa, Honduras. February/March 2001. PMID- 11852708 TI - The Charles Bernard Lecture. PMID- 11852709 TI - Sea-surgeons and the Company of Barbers and Surgeons. PMID- 11852710 TI - Impact of ADHD on children and their parents. PMID- 11852711 TI - The psychology of psychopharmacotherapy and the pharmacotherpeutic alliance. PMID- 11852712 TI - Effective management of adolescents with anorexia and bulimia. AB - 1. Anorexia and bulimia are common among adolescents, and both are complex, chronic disorders with high comorbidity and significant complications and mortality. 2. Eating disorders cultivate shame, denial, and lack of insight, making treatment difficult and necessitating careful development of a therapeutic alliance. 3. Treatment goals are to correct the biological and psychological sequelae, prevent or correct complications, restore normal eating patterns, reduce binging and purging, promote understanding and change in dysfunctional behaviors and attitudes, improve intrapersonal and interpersonal functioning, restore normal exercise patterns, and address comorbid psychopathology and psychological conflicts. PMID- 11852713 TI - Catapulting through life stages. When younger adults are diagnosed with life threatening illnesses. AB - Knowledge of developmental stages through the life cycle has always been a hallmark of quality nursing care. The knowledge base gleaned from the older adult literature, such as Schachter-Shalomi and Miller's construct of sage-ing (1995), can help nurses understand that many of the completion tasks usually associated with aging suddenly are thrust to the forefront for younger adult patients diagnosed with a life-threatening illness. Using this knowledge base, nurses can facilitate developmental stage work facing younger adult patients whose illness catapults them into more mature stages for which they may have been unprepared. When younger adult patients are so diagnosed, nurses need to recognize the signs of catapulting life stage work and support it. It is no small task to complete the gestalt of one's life tapestry, but it is especially difficult when one is young. PMID- 11852714 TI - Problems in community living identified by people with schizophrenia. AB - 1. Problems in community living were identified by a group of people with schizophrenia who received a telephone nursing intervention after discharge from a state psychiatric facility. 2. The most frequently reported problems were medication side effects, financial concerns, and anxiety. 3. A telephone nursing intervention has the potential to reduce the negative effects of stressors on people with schizophrenia living in the community. PMID- 11852715 TI - Poems, patients, and psychosocial nursing. AB - 1. Holism can prove just as elusive in psychosocial nursing as in any other specialty area. 2. Students in an undergraduate psychosocial nursing course used poetry to express the fullness of the situations in which they found themselves. 3. The poems touched a deeper and more complete sense of humanity than had been possible in other, more traditional, assignments. PMID- 11852716 TI - How could a mother do that? PMID- 11852717 TI - Obstructive sleep apnea: recognition and management considerations for the aged patient. AB - Sleep-disordered breathing (SDB) problems are estimated to affect between 2% to 4% of Americans with an even higher incidence occurring as one ages. This article focuses on obstructive sleep apnea (OSA) as a risk to safety and a leading contributor to interrupted sleep and disease morbidity in the aged population. Obstructive sleep apnea is caused by interruption in upper airway airflow during sleep that leads to oxygen desaturations and disruptions in sleep continuity. The symptoms, diagnosis, and initial management of OSA are discussed that provide direction for the advanced practice nurse who encounters patients with this disease. PMID- 11852718 TI - The challenge of managing the care of older heart transplant recipients. AB - Age is perhaps the most controversial exclusion criterion for heart transplantation. One concern focuses on whether chronological or functional age is the better predictor of positive outcomes when considering heart transplantation for an elderly patient with end-stage heart disease. Another concern is related to the philosophical and ethical rationale for allocation of scarce resources to those near the end of a normal life expectancy. However, the number of people who are older than age 65 years and have received a donor heart has increased and will continue to due to aging of the people who received a transplant a decade ago, as well as the growing number of people who undergo heart transplantation after the age of 65. In either case, the nurse must be aware of age-related concerns in this vulnerable population. PMID- 11852719 TI - An ethical analysis of cardiopulmonary resuscitation for elders in acute care. AB - Despite empirical evidence that cardiopulmonary resuscitation (CPR) is of questionable effectiveness in elders with comorbidities, it continues to hold a unique place in the armamentarium of life-sustaining treatments in that consent for CPR is implied and, when needed, is administered emergently. These conditions of implied consent and emergency implementation often preclude an opportunity for patients/surrogates, in conjunction with their nurses and physicians, to determine the appropriateness of the intervention, given the patient's medical and functional status. Healthcare providers who perform CPR on elderly patients often find themselves in morally distressing circumstances because of their anecdotal knowledge of poor outcomes and realization that a multitude of barriers has precluded an anticipatory discussion regarding appropriateness of the treatment. Nurses and other healthcare providers must take the lead in helping patients/surrogates appreciate the meaning of CPR as a life-sustaining intervention and ensure that each patient's situation receives the ethical reflection deserved for each individual as a matter of human dignity. PMID- 11852720 TI - Promoting positive outcomes for elderly persons in the hospital: prevention and risk factor modification. AB - The hospitalized elderly are at an increased risk for poor outcomes such as increased length of stay, readmissions, functional decline, and iatrogenic complications, as compared with other age groups. Research related to the hospitalized elderly has identified factors associated with poor outcomes. Nurses and other healthcare team members may be able to identify elderly patients at risk for poor outcomes and target modifiable factors to minimize their negative impact. Clinical experience and research validate the conclusion that multidimensional, preventive risk factor modification balanced with acute illness treatment can result in positive outcomes for elderly patients. PMID- 11852721 TI - Acute care of the elderly units: a positive outcomes case study. AB - Acute care of the elderly patient is a specialty area of practice. The elderly have a spectrum of needs including physiologic, psychosocial, functional, and financial needs that will alter their response to physiologic illness and the acute care experience in general. Acute care units specifically designed to meet the needs of elderly adults can have a dramatic impact on recovery from acute illness and long-term, postdischarge outcomes. This report describes the experience of one elderly patient hospitalized for the treatment of pneumonia. This elderly patient manifested a response to the acute care experience that was typical of his age cohort, including confusion and incontinence. Subsequent transfer to an acute care for the elderly unit maximized this patient's recovery and postdischarge placement. PMID- 11852722 TI - Treatment considerations for the elderly person with cancer. AB - In an aging population, the number of patients with cancer continues to rise. Little research has focused on the treatment of cancer in the elderly. Therefore, the treatment for various cancers differs across the healthcare system. A uniform approach in assessing the elderly person with cancer is lacking. This article describes two case studies in the elderly population, focusing on two common cancers: acute myelogenous leukemia and breast cancer. Common side effects of treatment and determinants of treatment options are discussed. It is important that the elderly receive appropriate screening, early detection, treatment, and management of comorbidities. PMID- 11852723 TI - HIV/AIDS in older adults: a case study and discussion. AB - Infection associated with human immunodeficiency virus (HIV) and acquired immunodeficiency syndrome (AIDS) in older adults is an increasing problem in clinical care. Often regarded as a disease of the young, more than 10% of HIV infection actually is found in people 50 years of age and older. In addition, individuals with HIV and AIDS are living longer. Approximately 71% of them currently are in their 30s and 40s. Given the current therapies available, it is conceivable that these patients will live well into their 60s and beyond. A case study describing the acute care experience of a 77-year-old African American man is reported. Pitfalls of diagnosis and management are discussed in relation to the care of an older person with HIV disease. The epidemiology of HIV in this population and a review of some recent literature and research on HIV and older adults are presented. PMID- 11852724 TI - Geriatric trauma: special needs for a special population. AB - Pediatric patients are not just "little adults" and elderly patients are not just "old adults." The elderly patient experiences physiologic and anatomic changes that affect all body systems. Providing trauma care for the elderly presents a particular challenge. Muscle atrophy, osteoporosis, and decreased subcutaneous tissue make the elderly patient more prone to a greater severity of injury. Alterations in the cardiovascular and respiratory systems limit the physiologic reserve the elderly need to respond to hypoxia and shock. Preexisting health conditions further complicate the picture. This article highlights some of the important differences in caring for an elderly trauma patient from resuscitation to rehabilitation. PMID- 11852725 TI - The challenge of assessing a patient with dementia and head injury. AB - Alzheimer's disease is emerging as a major health challenge for the 21st century. The reported case study discusses a 74-year-old woman with dementia of the Alzheimer type who sustained a head injury when she fell down the basement stairs. Differentiating the head injury from the preexisting dementia was complicated and required creative and astute assessment. Objective assessment tools discussed include the Mini-Mental State Examination, a delirium guide, and the Tinetti assessment tool. Predisposition to delirium is significant because of the comorbidities associated with cognitive impairment and head injury. Interventions to prevent delirium are recommended. PMID- 11852726 TI - Prolonged mechanical ventilation and tracheostomy in the elderly. AB - Mechanical ventilation is a life-sustaining technology used with increasing frequency in the elderly population. Prolonged mechanical ventilation is associated with high morbidity, mortality, and poor functional status. Care of these complex patients requires a coordinated multidisciplinary approach to optimize outcome. To minimize mortality and morbidity and contain health care costs, it is essential to identify patients at high risk for prolonged ventilation and to implement early interventions to curtail functional decline. In this article, the incidence and outcome of prolonged mechanical ventilation is reviewed, along with interventions to promote recovery. In particular, the role of tracheostomy timing and placement is discussed. PMID- 11852727 TI - Heart failure in older adults: keys to successful management. AB - Recent American Heart Association statistics indicate that approximately 5 million people experience heart failure, and that an estimated 400,000 to 700,000 new cases are expected annually. Improved efforts in the emergency care of myocardial infarction and the increased life expectancy of the population in general are credited for the rapidly increasing number of elderly adults with such chronic cardiac diseases as heart failure. New drug therapies are tested daily to improve the treatment of heart failure. However, drugs alone cannot improve the lives of elderly men and women with this disorder. Skillful delivery of expert care is necessary if advanced practice nurses are to reduce the burden of heart failure and improve the lives of the individuals who must live with this devastating disease. PMID- 11852728 TI - Arterial surgery facing the 3rd millennium. PMID- 11852729 TI - Treatment evaluation of hindfoot injuries caused by pressure activated explosive devices in the war and peace time. AB - Injuries caused by pressure activated explosive devices are very complex medical and social problem in Bosnia and Herzegovina. Hindfoot injuries are also a big challenge for orthopedic surgeons concerning all forthcoming physical assessment of the patients and the fact that the forefoot and midfoot remain preserved. The study included 60 patients with amputation that followed such an injury, and 60 patients treated with reconstructive procedures. The gait of the patients was assessed in the period of 4 years. In 20 patients, destroyed tissues were sent referred for pathohistologic examination, which revealed typical signs of a damage caused by high kinetic energy devices. The gait was analyzed objectively by the examiner and subjectively by the patients, which was the main indicator of successful treatment. Infection analysis and plantogram were also taken into account. The study showed that in a number of cases primary reconstruction of the hindfoot injuries caused by pressure activated explosive devices was possible with satisfactory results, thus foot amputation having strict indication. Indications for amputation have been limited and foot preservation has become more real in the majority of patients. PMID- 11852730 TI - Reed-Sternberg cells in the pathogenesis of Hodgkin's disease. AB - Hodgkin/Reed Stemberg (HRS) cells mediate the classical features of Hodgkin's disease. However, because of their rarity in tumor tissue, little is known about their origin and function. Recent advances in biotechnology, including the single cell manipulation, enabled the insight into the biology of HRS cell. It has been demonstrated that in the great majority of cases they are of germinal center B cell origin, with highly developed interactive network with adjacent cells via expression of cell adhesion molecules, tumor necrosis factor receptor superfamily, and elaboration of different cytokines. PMID- 11852731 TI - Antiphospholipid antibody syndrome and fetal outcome. AB - The presence of antiphospholipid antibodies, lupus anticoagulant and anticardiolipin antibody in patients with systemic lupus erythematosus has been associated with the clinical features of thrombosis, fetal loss and thrombocytopenia, and the syndrome is designated as antiphospholipid antibody syndrome (APS). APS has been increasingly diagnosed in patients without underlying autoimmune disease and is most frequently seen in obstetric patients suffering spontaneous abortion, preeclampsia and intrauterine growth restriction. The hypothesis underlying most research into the pathophysiology of APS is that autoantibodies are not only the markers of the disease, but also directly contribute to the development of clinical features. This review summarizes recent information on the pathophysiology and potential roles of autoantibodies in the obstetric patients suffering, particularly in the subgroup of repeated spontaneous abortions. PMID- 11852732 TI - Immunohistochemical and ultrastructural features of Gaucher's cells--five case reports. AB - Gaucher's disease is an autosomal recessive lysosomal storage disease resulting from glucocerebrosidase deficiency. In this report, five patients with adult Gaucher's disease are described. The clinical course of these patients was characterized by progressive diffuse aseptic necrosis in the large bones, so called Erlenmeyer's flask deformity, and hepatosplenomegaly. Splenomegaly was accompanied by hypersplenism with anemia and thrombocytopenia, therefore splenectomy was performed. The diagnosis of Gaucher's disease was based on the finding of Gaucher's cells on bone marrow biopsy. Tissue blocks were cut and routinely processed. Slides staining for iron (Peris' blue) and PAS (periodic acid--Schiff) including immunohistochemical staining for CD68 and HLA-DR was performed in all five cases. Gaucher's cells were seen as large cells with granular or fibrillar distended cytoplasm, with the characteristic 'wrinkled tissue paper' appearance, and eccentric nuclei. PAS staining showed strongly positive granular or fibrillar material in the cytoplasm. Immunohistochemical stain for CD68 and HLA-DR helped identify isolated Gaucher's cells, which are hystiocytic in nature. This stain accentuates their fine linear striations. Small pieces were ultrastructurally analyzed. PMID- 11852733 TI - Reconstruction of total lower lip, labial commissure and palatomaxillary defect with composite island cheek flap. AB - Functional and cosmetic restoration of total lower lip, labial commissure, and palatomaxillary defects can be achieved by using multiform flaps. The possibility of reconstruction of these areas with composite island cheek flap is presented. The flap was used in three patients who were surgically treated between 1993 and 1998. In one female patient, total lower lip and chin defect was restored with a small contralateral platysma muscle cutaneous flap. The functional and cosmetic result was achieved with composite island cheek flap. In the other two cases, reconstruction was performed without another flap. Composite island cheek flap is supplied with bloody by the facial artery and vein, and contains mucous membrane, muscle and skin of the cheek. In two cases, the flap was formed by the anterior part of the buccal muscle, and in the case with the reconstruction of labial commissure, the greater and lesser zygomatic muscles with levator muscle of the angle of mouth were used. By freeing the blood vessels from the surrounding structures, isolation and transsection of minor vessel branches allowed straightening of the tortuous main vessels. With this technique, excellent functional and satisfactory cosmetic results were achieved in a single act, with minimal morbidity for labial commissure, and total lower lip or palatomaxillary defect reconstruction. PMID- 11852734 TI - Extended-spectrum beta-lactamases (ESBL). PMID- 11852735 TI - Acute bulbar muscle dysfunction in hyperthyroidism. PMID- 11852736 TI - September 11, 2001: reflections of a trauma surgeon. PMID- 11852737 TI - Ghosts: living with the aftermath of managed care. PMID- 11852738 TI - Of genomics, cyborgs and nanotechnology: a look into the future of medicine. PMID- 11852739 TI - Improving care of diabetic patients through a collaborative care model. AB - The primary aim of this study was to evaluate the effectiveness of a hospital based diabetes disease management program. Effectiveness was measured in terms of glycemic control and adherence to American Diabetes Association (ADA) standards of care. Patients participating in the Diabetes LifeCare program received diabetes self-management education, medical management by a primary care provider (PCP) supported by an evaluation and recommendations by a Diabetes LifeCare Advanced Practice Registered Nurse (APRN), nutritional counseling and quarterly follow-up appointments through one year. There was a statistically significant decrease in HbA1c from 9.3 to 7.2 to 6.8. During a six-month follow-up period, adherence to ADA standards for annual eye examinations increased from 64% to 82% and from 66% to 85% for foot examinations as part of a physician visit. Significant improvement in glycemic control and adherence to standards of care can be achieved through a comprehensive program of patient education and management that includes collaborative efforts with the patient's primary care provider. PMID- 11852740 TI - Functional health status of persons with diabetes in a nurse-managed clinic. AB - PURPOSE: The purpose of this psychometric analysis was to (1) describe the functional health status of medically underserved persons with diabetes participating in an exercise program; (2) examine the relationships between functional health status scores and physiologic measures for blood pressure, heart rate, and blood glucose levels; and (3) examine the reliability and validity of the Medical Outcomes Study SF-36 survey. METHODS: Data were collected from 31 men and women receiving diabetes disease management in a community-based nurse-managed clinic. The SF-36 survey was used to measure functional health status. Blood pressure, heart rate, and blood glucose also were measured (pretest/posttest). RESULTS: The SF-36 survey had good internal consistency in the total sample. Blood pressure and blood glucose levels decreased significantly and heart rate increased significantly from pretest to posttest. Functional health status was negatively associated with physiological parameters. CONCLUSIONS: Further research is needed on the use of the SF-36 survey in medically underserved persons with diabetes because this instrument lacked construct validity as a measure of functional health status in this population. PMID- 11852741 TI - Increasing problem solving in adolescents with type 1 diabetes: the choices diabetes program. AB - The purpose of this pilot study was to test the hypothesis that adolescents with type 1 diabetes can learn to become better problem solvers in diabetes self-care and thereby improve their metabolic control. METHODS: Fifty-three adolescents aged 13 to 17 with type 1 diabetes were randomly assigned to either a 6-week problem-solving diabetes education program or to a control group (usual care). A1C levels were obtained as well as assessments of problem solving, frequency of behavior, level of responsibility, and 24-hour behavior recall at baseline and 6 months. RESULTS: The experimental group participants showed significantly improved problem-solving test scores and A1C values from baseline to 6 months, changes not evident in the control group. At 6 months, the experimental group participants were doing blood glucose testing more often than those in the control group. However, there was no significant difference in problem-solving test scores or A1C values. CONCLUSIONS: This 6-week intervention for adolescents with diabetes resulted in better problem-solving skills, more frequent blood glucose testing, and improved A1C values. The results suggest that a diabetes problem-solving program for adolescents can be effective in improving metabolic control. PMID- 11852742 TI - Important considerations of the patients' bill of rights. PMID- 11852743 TI - Assessing diabetes patients' healthcare needs. AB - Pharmacists who possess the proper skills and knowledge can intervene with patients in a proactive manner by initiating a dialogue at the point of dispensing medication. After patients complete the DPAQ and the pharmacist assesses the information, a plan of care is developed. Proper documentation (e.g., using the SOAP format) is the first and most important step in caring for patients with diabetes. Such encounters impact on the level of care provided and are professionally and financially rewarding for the pharmacist. PMID- 11852744 TI - A comparison of insulin pen use in the United States and the United Kingdom. AB - Insulin pens offer the advantages of simplicity, convenience, and more accurate dosing to insulin-using patients with diabetes. The usefulness of insulin pens is not limited to certain subsets of individuals but extends to all patients who might choose this delivery system. By facilitating acceptance and consistent implementation of multiple-dose insulin regimens, pens hold the potential to promote improved blood glucose control and thus reduce the risk of the chronic complications of diabetes. Both clinical experience and the supporting literature suggest that pen delivery systems are an option that should be routinely offered to all insulin users. PMID- 11852745 TI - Nutrition interventions in early diabetic renal disease. AB - Education and nutritional intervention play an important role in preventing and delaying the progression of renal disease in persons with diabetes. We need to identify those patients at risk or who have renal disease early, when interventions are most effective. Patients should visit a dietitian with expertise in diabetes to obtain an individual meal plan that focuses on those aspects that have the greatest impact on the disease: blood pressure control, blood glucose control, and moderate protein restriction. PMID- 11852746 TI - Toward a better understanding of diabetic nephropathy. PMID- 11852747 TI - Syringe disposal practices and gender differences. AB - PURPOSE: The purpose of this study was to describe syringe disposal practices of individuals with diabetes who take insulin and to investigate the attitude toward and effect of previous information on proper syringe disposal. METHODS: Adult men and women who injected insulin were recruited from an endocrinology practice and given 2 questionnaires concerning syringe disposal practices and attitude toward proper syringe disposal. Completed questionnaires were returned by mail to the investigator. Nonparametric statistical procedures were used to analyze the data. RESULTS: Men with higher income tended to use the trash for syringe disposal more often than lower income men, and older women exhibited a higher positive attitude score than younger women. A positive attitude significantly correlated with the likelihood of proper syringe disposal, and those who had received previous information were more likely to dispose of syringes properly. Healthcare providers were identified as the main source of information. CONCLUSIONS: Diabetes educators can improve the syringe disposal practices of their patients through education at each office visit. Knowing more about the environmental and social problems that may ensue from improper syringe disposal may influence patients to practice better disposal methods. PMID- 11852748 TI - Do persons with diabetes know their (A1C) number? AB - PURPOSE: The objective of this study was to compare self-reported knowledge about A1C testing with information from the medical record. METHODS: A telephone survey was conducted among patients with diabetes in a rural fee-for-service practice and a community health center. Self-reported information regarding A1C testing, the last A1C value, and perceived blood glucose control was compared with the most current A1C value documented in the medical record. RESULTS: Seventy five percent of survey respondents reported having 1 or more A1C tests in the past year, which generally agreed with information from their medical records. However, only 24% of those who reported having a test remembered the actual value, and the self-reported values correlated weakly with the last A1C on the medical record. Among those with a documented A1C value, half described their blood glucose as very well controlled. The last A1C value, however, was < 7.0% in only half of those respondents. CONCLUSIONS: Persons with diabetes were aware of their previous A1C testing but did not interpret the values accurately in relation to their own glycemic control. If clinicians expect patient knowledge and understanding of glycemic control measures to improve outcomes of care, patient education will need to emphasize the meaning of these values. PMID- 11852749 TI - Trained nurses in family magazines, 1880-1928. PMID- 11852750 TI - Nurses: the early twentieth century tuberculosis preventorium's "connecting link". PMID- 11852751 TI - The roots of collaborative practice: nurse practitioner pioneers' stories. PMID- 11852752 TI - The fork in the road: nursing history versus the history of nursing? PMID- 11852753 TI - The wind of change is blowing. PMID- 11852754 TI - To work in the Garden of God. The Swedish Nursing Association and the concept of the calling, 1909-1933. PMID- 11852755 TI - The beginning of nursing in Brazil. Brazilian sanitarians and American nurses. PMID- 11852756 TI - "The problem" of student nurses of Japanese ancestry during World War II. PMID- 11852757 TI - Caring for life: nursing during the Holocaust. PMID- 11852758 TI - Smaller and cheaper: the Chicago Hourly Nursing Service, 1926-1957. PMID- 11852759 TI - [Functional anatomy and semiology of the bladder]. AB - Continence and micturition result from an inversely related evolution of urethra and bladder pressures. On filling, the bladder pressure remains low and the urethral pressure is high: filling bladder pressure mainly depends on its visco elastic properties while urethral pressure is actively kept high by the tonic activity of smooth and striated urethral sphincters. It is the other, way round when the bladder empties: bladder pressure rises and urethral pressure lower, because the parasympathetic impulses; in the adult, the reflex loop passes through the pontine center. The volitional control of this micturitional reflex implies an ability to recognize a specific feeling: the desire to urinate. Physical examination and investigations enables to assess bladder and sphincter function, through a rich semiology. PMID- 11852760 TI - [Acute bacterial cystitis]. AB - It is now important to differentiate lower urinary tract infection also known as classical acute cystitis (because it involves a healthy genitourinary system) from complicated or potentially complicated urinary tract infection. The bacteriology of lower urinary tract infection is dominated by enterobacteria and Escherichia coli in particular. Since E. coli are now commonly resistant to amoxicillin and cotrimoxazole, those antibiotics cannot be used as first intention. Fluoroquinolones, fosfomycin trometamol or furans must be preferred instead. The clinical problem here is trying to avoid overzealous diagnoses and remembering that symptom-free bacteriuria is very frequent, particularly in elderly patients. The management of uncomplicated lower urinary tract infection should remain simple (urinalysis with reagent strips and more cell bacteriology tests); the treatment should be simple too [single-dose or short treatment (3 days)]. The management of complicated lower urinary tract infection is very different however and requires a more extensive workup followed by a dual therapy, namely antibiotics and eradication of the complicating factor, if possible. PMID- 11852761 TI - [Interstitial cystitis]. AB - Interstitial cystitis is a disease characterized by pain occurring as the urinary bladder is gradually filling up. Its pathophysiology is unknown. The cause could be either a membrane permeability disorder, a mast cell disturbance in the bladder wall, or an autoimmune process involving the bladder. The diagnosis is essentially clinical but endoscopies and biopsies are required to rule out vesical cancers. Treatments is difficult and based on a combination of sanitary and dietary regimens, as well as drugs administered orally or infused within the bladder. Surgical treatment must remain exceptional. PMID- 11852762 TI - [Diagnosis of bladder tumors]. AB - Prompt diagnosis of bladder cancer is essential to improve the management of this disease. Hematuria and lower urinary tract symptoms must draw attention to bladder difficulties. The diagnosis is based on cystoscopy and biopsy of any lesions. Once the diagnosis has been established, the tumour should be staged using the TNM system before treatment. Research efforts are continuing to identify biological markers sensitive and specific enough to predict the diagnosis and behavior of this disease accurately. PMID- 11852763 TI - [Treatment of bladder tumors]. AB - The bladder tumour treatment closely depends on initial diagnosis. For superficial bladder tumour a conservative treatment is required using endoscopic resection. Adjuvant instillations (BCG or mitomycine C) can be proposed for high grade tumour. In case of muscular layer invasion or in case of frequent non controlled recurrence, cystectomy is necessary. Progress in anaesthesia, surgery and postoperative care permitted to enteroplasty to be much less morbid, it must be considered as the urinary diversion of choice. To prevent recurrence, both (superficial or infiltrating) tumour require very strict follow-up for many years. PMID- 11852764 TI - [Incontinence and voiding disorders]. AB - Urinary continence and micturition are functions which require the integrity of the organs (bladder, urethra, voluntary and involuntary sphincters) and the neural pathways responsible for micturition (parasympathetic), continence (sympathetic), and their control and coordination. Save the incontinence associated with vesico-vaginal fistulae in women, or overflow incontinence associated with a distended bladder in chronic retention, the three principal clinical forms of incontinence are: stress incontinence, urge incontinence and mixed incontinence, combining the two mechanisms. Voiding difficulties causing discomfort on urination, or even retention (chronic or acute), are the reflection of an imbalance between bladder contraction (reduction) and urethral resistance (augmentation). The management of a voiding disorder systematically requires an understanding of the pathophysiological mechanism involved and the search for the aetiology (often multifactorial), so as to adapt the appropriate medical and/or surgical management. PMID- 11852765 TI - [Cystocele]. AB - A cystocele is a bladder hernia through the anterior wall of the vagina. It is common in elderly women. There is no clinico-anatomical correlation, but the functional disturbance, although both variable and variably experienced, remains a major factor in the indication of eventual surgery. Methodologic clinical examination should be made to seek anomalies of pelvic stasis associated with a more or less hidden urinary incontinence. The surgical technique should be chosen according to the age and the classification of the patient, determined by complementary examinations that range from simple to highly technical (echography, cystography and sometimes study of urinary dynamics and MRI). The surgeon should chose the technique of perineal support and clearly explain the possibility of failure or recurrence. Confidence between the patient and the surgeon remains the best guarantee of a good result of management. Such surgery can be proposed to women to request it for physical comfort, whereas the surgeon should be very cautious with regard to those who do not want to undergo surgery (but also with those who strongly insist on surgery). PMID- 11852767 TI - [The catheterized bladder]. AB - Whatever the cause is, the retention of urine needs a bladder drainage whose mode must not be considered as an evidence. Otherwise, the immediate easiness is likely to be the only criterion of choice. The indwelling catheterization is thus overused, whereas its complications (infectious, cancerous, patient comfort...), which are particularly frequent, should make it be perceived secondarily difficult to manage. Its alternatives, particularly clean intermittent self catherization and suprapubic catheter, have to be first and systematically discusseed, as well as the relevance of the bladder catherization itself. PMID- 11852766 TI - [Migraine: a disease, not a symptom]. PMID- 11852768 TI - [Medical demography: state of localities and outline of prospects]. PMID- 11852769 TI - [Multiple myeloma of the bone]. PMID- 11852770 TI - [Myasthenia gravis]. PMID- 11852771 TI - [Ectoparasitic skin diseases. Scabies and pediculosis]. PMID- 11852773 TI - [Siamese twins in classical Antiquity: from myth to fair phenomenon]. PMID- 11852772 TI - [Main complications of pregnancy. Fever and pregnancy]. PMID- 11852774 TI - [Tumors of the esophagus]. PMID- 11852775 TI - [Heart failure in adults]. PMID- 11852776 TI - [Acute hepatitis C in the presence of chronic HBV infection]. AB - Acute HCV superinfection was studied in 23 patients with chronic hepatitis B virus infection. HBsAg, anti-HCV (C-100, core, NS3, NS5) were detected in patients' sera at first investigation. Predominant replication of HBV DNA was detected in the sera of 68% patients and HCV RNA in only 24% patients. The clinical course of acute hepatitis C in patients with chronic HBV infection in general corresponded to HCV monoinfection except for more pronounced biochemical shifts and shorter intoxication. The role of HBV and HCV in infectious process is discussed. PMID- 11852777 TI - [Preparation and selection of monoclonal antibodies for detecting hepatitis B surface antigen in blood sera]. AB - HV monoclonal antibodies (MAb) were produced in order to improve the quality of HBsAg detection and their specific characteristics were compared with those of other MAbs. MAbs were characterized by asymmetric interactions with the antigen when used as first or second antibodies. The reactivity of a panel of HV and X MAb to ad and ay subtypes was studied by enzyme immunoassay. Mutual blocking (epitope mapping) of MAb helped select antibody couples for the creation of highly effective test system for the diagnosis of the major HBsAg subtypes. The sensitivity and specificity of MAbs were evaluated on reference and control panels of HBsAg sera and on serum specimens from a random sampling of 300 blood donors. The sensitivity of the most specific MAb pairs was 0.1 ng/ml for HBsAg subtype ay and 0.25 ng/ml for subtype ad. The specificity of attested MAb was 98.5% in incubation with stirring and 97% in static incubation. The optimal combinations of attested MAbs were used in the manufacture of Recomnathep B test system in the sandwich format. PMID- 11852778 TI - [Characterization of a panel of monoclonal antibodies to hepatitis C NS3 recombinant protein ]. AB - Recombinant protein rNS3 imitating helicase region (1356-1459 amino acid residues) of hepatitis C virus (HCV) was expressed in E. coli cells and used for BALB/c mice immunization. Seven hybrydoma clones producing monoclonal antibodies (MAbs) to rHS3 were obtained. All MAbs reacted in ELISA with NS3 protein from Murex anti-HCV Version III and in immunoblotting from RIBA 3. These MAbs detect 5 individual epitopes, 4 of which were conformational and 1 discontinuous. All MAbs could compete for rNS3 binding with serum antibodies from patients with chronic hepatitis C, which suggests that these MAbs can recognize the natural HCV NS3 protein. PMID- 11852779 TI - [Combined experimental infection of Microtus gregalis by tick-borne encephalitis and rabies viruses and probable interference]. AB - Narrow-skulled voles (Microtus gregalis) were inoculated with tick-borne encephalitis (TBE) and rabies (RV) viruses isolated in the steppe zone of West Siberia. TBE infection was asymptomatic and nonlethal in all voles. The virus was eliminated from the voles between days 14 and 60 postinoculation. Only 1 of 26 tested animals produced anti-TBE antibodies in titer 1:20. RV infection was acute and lethal. All sick animals contained RV in the brain, 42.1% in salivary glands, 5.3% in the lungs and kidneys. The voles which survived RV inoculation had no RV. The voles inoculated with RV after TBE were less sensitive and had a longer incubation period in comparison with the animals challenged with RV alone. The differences augmented from day 0 to day 14 after TBE inoculation and became statistically significant on day 14. Later, 21 day after challenge with TBE virus, the differences almost disappeared. The role of possible interference in the evolution of virus-host interaction of Lyssaviruses is discussed. PMID- 11852780 TI - [Suppression of cytomegalovirus infection in cell system by fullerene amino acid derivates]. AB - Effects of two water-soluble derivatives of fullerene C60-o-aminocaproic acid (C60-ACA) and C60 sodium salt of omega-aminocaproic acid (C60-Na-ACA) on in vitro cytomegalovirus (CMV) infection were studied. C60-Na-ACA 4-5-fold inhibited the cytopathic effect of CMV in comparison with C60-ACA, the effective dose for C60 Na-ACA being 0.6 microgram/ml and that for C60-ACA 2.7 micrograms/ml. Immunocytochemical analysis of virus proteins in infected cells has shown that C60-Na-ACA inhibits the production of late structural CMV protein gB, but does not modify the expression of immediate early nonstructural protein IEp72. Studies of cell viability, growth characteristics, and DNA synthesis revealed that the cytotoxic effect of C60-Na-ACA on human diploid fibroblasts in negligible, the cytotoxicity index varying from 160 to 1500 micrograms/ml in different tests. Selectivity index for C60-Na-ACA is 267-2500, which differs negligibly from that of gancyclovir (100-1000), while the cytotoxicity of C60-Na-ACA is essentially lower. PMID- 11852781 TI - [Therapeutic effect of gefin, a new antiviral drug, in experimental genital herpes ]. AB - Antiviral activity of Gefin was studied in guinea pigs infected with herpes simplex virus (HSV-2). Up to 54.9% animals challenged with HSV survived after 7 day treatment with Gefin (3 local applications a day). The prospects of further trials of the drug in HSV genital herpes are discussed. PMID- 11852782 TI - [Effects of hyaluronic acid preparation on the development of herpetic infection in cell culture]. AB - Cell culture experiments demonstrated antiviral activity of a new hyaluronic acid preparation towards HSV-2. The active concentration of hyaluronic acid is at least 5%. PMID- 11852783 TI - [Combined vaccine protecting from transmissible swine gastroenteritis]. AB - An effective system for specific prevention of transmissive swine gastroenteritis (TSG) is based on the use of combined vaccine and combined vaccination. Combined vaccine is a lyophilized preparation for intranasal application and emulsion for intramuscular injection. The former is prepared from natural attenuated respiratory variant of TSG virus and the latter from artificially attenuated TSG virus. Combined vaccination consists in parallel double immunization of females on days 70-75 of gestation intramuscularly and intranasally and on days 90-95 of gestation intramuscularly. Vaccinal lactogenic immunity ensures protection of newborn piglets under experimental and field conditions (protection of 70-80 and 90-95% piglets, respectively). The proposed vaccine preparations and protocols of combined immunization of females during gestation by two methods are an effective and safe method for specific prevention of TSG. PMID- 11852784 TI - [HIV-1 chemokine receptors and their role in the pathogenesis of AIDS]. AB - Data on cellular receptors playing an important role in penetration of HIV into the cell and determining further development of HIV infection are reviewed. Special attention is paid to chemokine receptors CCR5 and CXCR4. The resistance of some humans to HIV-1 infection, probably due to defective CCR5 gene allele, is discussed. PMID- 11852785 TI - [Cytokine production by blood cells in herpes, hepatitis C, and other diseases]. AB - A new method for simultaneous evaluation of interferon and cytokine production by human blood cells is developed. Study of interferon and cytokine status helps evaluate the possibility of correction of the detected disorders by immunomodulating agents. PMID- 11852786 TI - [Probable role of viral infections in mental diseases of man (hypotheses and facts)]. PMID- 11852787 TI - [Genome analysis of hepatitis C virus strain 274933RU isolated in Russian Federation]. AB - Five overlapping cDNA fragments of hepatitis C virus (HCV) isolate 274933RU, obtained by RT-PCR, were amplified and cloned. Complete nucleotide RNA sequence has been determined. The genomic organization of 274933RU was, from 5' to 3' terminals, 5' UTR (341 nt), polyprotein ORF (9033 nt), 3' UTR (40 nt except for the poly(U-UC) and polypyrimidine stretch), and X-tail (98 nt). Phylogenetic analysis of the core and NS5 genes showed that the isolated strain belonged to HCV 1b subtype. PMID- 11852788 TI - [Changes in the frequency of alleles and genotypes of insertional-deletional polymorphism of the angiotensin-converting enzyme gene, dependent on age]. AB - Insertion-deletion polymorphism of the gene for the angiotensin-converting enzyme has been investigated in random samples from various age groups in the Moscow population. A statistically significant reduction in the insertion allele frequency has been found in senior age groups. PMID- 11852789 TI - [CG17604 gene from Drosophila melanogaster--possible functional homolog of the yeast ZIP1 and SCP1 (SYCP1) mammalian genes, coding for synaptonemal complex proteins]. AB - From data on the molecular organization of transverse filament proteins of the synaptonemal complex (SC)--Zip1 in yeast and SCP1 in mammals--and on the width of the central SC space in these organisms and in Drosophila, the putative molecular structure and size of a transverse filament protein of the SC in Drosophila has been inferred. Using genetic and molecular databases and software from the Internet, we carried out in silico screening for a candidate gene for the Drosophila transverse filament protein. The search in the 250-bp region overlapping the locus of this gene (sections 88E-89B) and containing 78 predicted genes has revealed only one gene, CG17604, whose protein meets all requirements for the transverse filament protein of the SC. It was suggested that gene CG17604 is gene c(3)G. In this case, gene c(3)G must be localized in section 89A7-8 of the cytological map of Drosophila melanogaster. PMID- 11852790 TI - [Characteristics of genes identified in the 120 MDa plasmid DNA in a mutant of Azospirillum brasilense Sp245 bacteria, defective in polar flagellation and swarming]. AB - The sequencing data were analyzed for two regions of the 120-MDa plasmid (p120) of Azospirillum brasilense Sp245. The 2420-bp region I, which flanks the omegon insertion in the SK048 mutant defective in production of the polar flagellum (Fla ) and swarming (Swa-), was shown to contain a cluster of two open reading frames (orf) that possess properties of coding sequences (CDSs). The NtrA (sigma 54) boxes were found in their upstream regions. The products of orf1 and orf2 are 16.5 and 15.5 kDa in molecular weight and consist of 151 and 152 amino-acid residues, respectively. The PRF1 polypeptide was found to contain a region homologous to the cysteine- and glycine-rich zinc-binding domain of the DnaJ heat shock protein. ORF2 showed a homology to Haemophilus ducreyi pilin, fragments of Streptomyces and Mycobacterium integral membrane proteins, and eukaryotic transcriptional regulators. The omegon proved to be inserted into orfX1/X2 which possibly has a deletion and shows a GC content untypical for A. brasilense genes. The deduced ORFX2 polypeptide is homologous to fragments of arsenite translocating ATPase and signal-transducing histidine kinase of archaebacteria. Possible causes of the Fla-Swa- phenotype of the A. brasilense SK048 mutant are considered. One coding orf was identified in the 1194-bp region II located approximately 4 kb away from the omegon insertion. The N-terminal region of the deduced product of this partly sequenced orf was shown to contain a signal sequence typical for secreted proteins. PMID- 11852792 TI - [Identification of Indy(p115) mutation in the Na+-dicarboxylate transporter gene in Drosophila melanogaster]. AB - The P[lArB]-element insertional mutation (Indyp115) was obtained in the promoter region of the Na(+)-dicarboxylate transporter gene of Drosophila (gene Indy, I'm not dead yet) within the 75D region of chromosome 3. The expression pattern of the reporter beta-galactosidase was determined in various tissues of third-instar larvae and adult flies. Both males and females homozygous for this mutation were fertile, though their viability was reduced. The two lethality phases were revealed in embryogenesis at nucleus cleavage stages 1-5 to the point of blastoderm formation and at latest stages 15-16, as well as at stage 17, when the larval development is completed though the larva still remains in the chorion. The gene expression in the follicular cells of an embryo at the terminal oogenesis stages is suggested to cause the first lethality stage. The expression pattern of this gene seems also to account for the tissue- and stage-specific activity of the 5'-regulatory region in the Indy gene. PMID- 11852791 TI - [Detection of two polymorphic sites in the human c-fms gene: allele frequency and genotype in some populations of Russia]. AB - Two polymorphic sites were found in the human c-fms gene: one (G-->A) was in position 34,047 in the last intron, and the other (dinucleotide TC-->CA) was in positions 34,293 and 34,294 in the 3'-untranslated gene region, 34 bp downstream of the translation stop codon. The polymorphic dinucleotide appeared to be immediately upstream of an octamer showing 100% homology to cis element -CAAACTTC , which is responsible for controlled instability of mRNAs of several genes. Based on these data, functional significance was assumed for this polymorphism of the c-fms gene. Allele frequencies were established for several populations. The mutant allele of the polymorphism located in the intron were detected only in one family of ethnic Germans from the Altaiskii krai. Polymorphism of the second site, which is in the 3'-untranslated region of the c-fms gene, was observed in all Caucasoid and Mongoloid populations examined. Frequency of the rare allele varied from 19.7-25% in Arctic Mongoloids to 31-42.6% in Central Asian Mongoloids and was similar in two Caucasoid populations (22.6% in ethnic Russians and 26.5% in ethnic Germans). The wide distribution of the mutant allele in human populations of the two races was considered indicative of an adaptive role of the polymorphism in providing a certain level of the gene product, a receptor, in certain cell processes. PMID- 11852793 TI - [Induction of MGE 412 transposition individually by heat and cold shock in spermatogenesis in Drosophila males]. AB - Effects of temperature treatment (heavy heat shock, HHS; heat shock, HS; and cold shock, CS) on the daily productivity of treated males in different spermatogenesis stages have been studied in isogenic line 51 of Drosophila melanogaster. The average productivity was shown to substantially decrease in all cases. The sum of the HS and CS contributions to this decrease was nearly equal to the HHS (the combined HS and CS) contribution, i.e., these contributions were almost additive. The temperature treatments did not kill mature sperm. In the control, mating productivity of day 1 exceeded that of the next day at least by 10-20%. Each day, most sperm in matings was new, i.e., matured during that day. Transposition induction of MGE 412 was studied at four spermatogenesis stages after HS and CS. Both temperature treatments were effective but CS had a more pronounced inducing effect. Most temperature-induced transpositions occurred at stage 3 (meiosis) and 4 (spermiogenesis). The day rates of transpositions at different stages were estimated. After HS at the meiosis stage, lambda = 0.11 events per initial MGE copy per sperm per day of mating, which is approximately equal to the previous estimates after HHS. After CS at the meiosis stage, lambda = 0.51. The transposition hot sites (including the previously known 43B and 97DE as well as a number of new sites) were detected. The lists of transpositions after CS completely included the corresponding lists after HS, which suggests similarity of induction mechanisms underlying CS and HS. PMID- 11852794 TI - [Transgenic goats in the world's pharmaceutical industry in the XXI century]. AB - In many developed countries, isolation of human pharmaceutical proteins from milk of genetically modified animals is currently a priority. One of the first commercial pharmaceuticals obtained from the milk of transgenic goats, an anticoagulant antithrombin III, developed by Genzyme Transgenic Corporation, an American biotechnological company, will appear on the pharmaceutical market in the nearest future. In this review, we discuss the role of fundamental science in the development of this field of the pharmaceutical industry. PMID- 11852795 TI - [Inheritance of reproductive traits of medicinal leeches (Hirudo medicinalis L.)]. AB - The phenotype variability and inheritance of reproductive traits were investigated in the medicinal leech. Distribution parameters were determined for the following traits: batch size (X = 4.3 +/- 0.2, sigma = 1.7, CV = 40%, As = 0.23 +/- 0.25, Ex = 0.19 +/- 0.51), number of juveniles in a cocoon (X = 10.9 +/- 0.3, sigma = 4.6, CV = 42%, As = 0.31 +/- 0.15, Ex = 0.23 +/- 0.30), and juvenile weight (X = 32.0 +/- 0.3, sigma = 14.9, CV = 47%, As = 1.38 +/- 0.05, Ex = 3.32 +/- 0.11. A nonlinear negative correlation between the number of juveniles in a cocoon and their weight was found (correlation ratio R = 0.86). It was shown that the environmental variance dominated over the genotypic one in the structure of phenotypic variance of the traits studied. The genetic variability is determined mainly by additive gene interactions and, to a small extent, intralocus dominance. The narrow-sense heritability, h2, for batch size was 0.35-0.40; for the number of juveniles in a cocoon, 0.35; for juvenile weight, 0.42. PMID- 11852796 TI - [Nana gene--a regulator of cell division and elongation of stem cells in Arabidopsis thaliana (L.) Heynh]. AB - The dominant nana (na) mutation localized to the upper arm of Arabidopsis thaliana chromosome 1 blocks cell proliferation in apical meristem (AM) of the flower-bearing stem at its early development and suppresses the subsequent elongation of its internode cells. The na mutation reduces the sensitivity of cells of the flower-bearing stem to gibberellin (GA) and paclobutrazole (PBZ) and prevents resting and immature seeds from restoring the germinating ability in response to exogenous GA. On the other hand, exogenous GA and PBZ affects the onset of flowering, hypocotyl length, and leaf color; i.e., the na mutant displays a distortion of only several, rather than all, GA-dependent processes. Based on the results obtained, the product of the NA gene was assumed to play a role in the negative regulation of GA signaling and to act later than the products of the known GAI and SPY genes. PMID- 11852797 TI - [Variability of the number of chloroplasts in a population of stoma guard cells in the sugar beet (Beta vulgaris L.)]. AB - Diploid sugar beet plants demonstrate a broad variability of the number of chloroplasts in stoma guard cells, which is related to myxoploidy of cell populations in leaf apical meristems (epigenomic variability). In addition to random organelle segregation between daughter cells, this variability is affected by factors disrupting the mitotic cycle: (1) plant treatment with a mitotic poison, such as colchicine; (2) duration of the life cycle of a plant; the variability in second-year plants is greater than in first-year ones; (3) the mode of plant reproduction; the variability in inbred plants is greater than in the initial population. Treatment of germinating seeds with a diluted colchicine solution increases the number of organelles in cells in the myxoploid generation (generation C0) and the variance of the distributions in the first vegetation year. The variability in the organelle number in stoma cells correlates with that in maternal meristem cells. It is concluded that the epidermal cell monolayer (including stoma guard cells) keeps record of the epigenomic and epiplastome variability in meristem cells. The variability of the number of chloroplasts in stoma guard cells is approximated by binomials with negative powers. PMID- 11852798 TI - [Intraspecies phylogenetic connections in Sorex araneus L.: a southern Baltic subgroup of chromosome races]. AB - Results of chromosome G-banding were used to identify two new karyotypic races of the common shrew (Sorex araneus L.) in the European part of Russia. The chromosomal diagnosis of race St. Petersburg included five metacentrics (hk, ip, jl, mq, and nr) and two acrocentrics (g and o) (2Na = 20); the diagnosis of race West Dvina included six metacentrics (gm, hk, ip, jl, no, and qr) (2Na = 18). The phylogenetic significance of the chromosomal markers gm, hk, and ip is considered and the possibility of reticulate evolution in the species S. araneus is discussed. PMID- 11852799 TI - [Genetic structure of a diploid-polyploid complex of the spined loach Cobitis taenia (Cypriniformes: Cobitidae) from the middle Dnieper bassin]. AB - Biochemical genetic typing and cytometry showed that polyploid females account for 87% of the spined loach Cobitis taenia population from the middle Dnieper basin. The polyploidy series included triploids, tetraploids, and, possibly, a few pentaploids. A characteristic feature of the genetic structure of polyploids was that their genetic variation was due to the clonal variation in the haploid portion of the genome originating from Cobitis sp. and to polymorphism of the diploid portion originating from C. taenia. The results are discussed with regard to comparative evolution of alloploid complexes in fish and terrestrial vertebrates. PMID- 11852800 TI - [Comparison of Chuvashs with Maris and Russians by vital statistics and the Crow index]. AB - Genetic demographic characteristics were calculated for Chuvash and Russian inhabitants of the Republic of Chuvashia. The generation lengths were 27.09 and 26.4 years and the sibship sizes were 2.54 and 1.82 for Chuvashes and Russians, respectively. Crow's indices and their components were as follows: Im = 0.05, If = 0.31, and Itot = 0.37 for Chuvashes and Im = 0.03, If = 0.43, and Itot = 0.46 for Russians. The genetic demographic characteristics obtained were compared with those for Highland and Meadow Maris. PMID- 11852801 TI - [Polymorphism of the STR-locus of Y chromosomes in Eastern Slavs in three populations from Belorussia, Russia and the Ukraine]. AB - Allelic polymorphism of five microsatellite loci of the human Y chromosome (DYS19, DYS390, DYS391, DYS392, and DYS393) was analyzed in samples of male populations from Ukraine, Russia, and Belarus (152 subjects in total). The allelic diversity indices (Dg) were determined for all loci; they varied from 0.23 to 0.72. The mean values of this parameter in the Ukrainian, Russian, and Belarussian populations were 0.45, 0.47, and 0.52, respectively. A total of 53 different haplotypes were found in 152 subjects from three populations. The most frequent haplotype was found in 14.5% of the subjects, whereas 35 haplotypes (23%) were each found in only one person. The haplotypic diversity index (Dhp) was 0.94. The genetic distances between the populations studied and some populations of Western and Central Europe were estimated. These data were used to construct a phylogram (tree) of genetic similarity between the populations, which demonstrated that the three Eastern Slavic populations are genetically close to one another and remote from Western European populations. PMID- 11852802 TI - [Cytokines: regulations of immune response during infection]. AB - The aim of this work is to review state-of-the-art knowledge about cytokines and their role as regulators of immune response during infections caused by different pathogens. PMID- 11852803 TI - [Preliminary estimation of results of high-dose intravenous immunoglobulin in severe steroid-dependent adult asthma]. AB - The aim of this study was to estimate efficacy and safety the high-dose intravenous immunoglobulin's (IVIG) in severe steroid-dependent adult asthma. The study group consisted of 10 asthmatic patients aged 18-74 (in an average 53 +/- 15) who received 6 months infusions therapy (7 doses). Each IVIG dose was 2 g/kg/month of body mass. In treatment monitoring the following parameters were used: asthma score, drugs (systemic and inhaled corticosteroids, beta 2-mimetic) use, daily PEF and monthly FEV1. Preliminary estimation of the obtained results shows the reduction of systemic corticosteroids doses and simultaneous improvement of asthma score and FEV1 as well as the decrease of asthma aggravation and emergency need among examined patients. Basing on this experience in high-dose IVIG treatment of patients with severe steroid-dependent asthma one could conclude about the safety and effectiveness of this method, particularly about its influence on the reduction of corticosteroids doses necessary for asthma control. The short time of observation does not allow to estimate the persistence of therapeutic effect after finish of this therapy. However, in some patients the recovery persisted for 6 months after the ending of IVIG infusions. PMID- 11852804 TI - [Selected parameters of the oxidative-antioxidative balance in children with bronchial asthma exacerbations]. AB - The aim of this study was to evaluate the involvement of selected parameters of oxidative-antioxidative system in children with bronchial asthma exacerbations. In my study the intensity of lipid peroxidation was calculated by the contents of malolonic dialdehyd in plasma. In order to evaluate activity of antioxidative system I investigated vitamin E concentration and total antioxidants status in serum, as well as activity of glutathione peroxidase and superoxide dismutase in blood. My study was performed on 32 children treated for exacerbations of bronchial asthma. The control group consisted of 27 healthy children. The study has revealed that children suffering from bronchial asthma exhibit disturbances in the oxidative-antioxidative balance, manifested by increased oxidative activity and by changes in certain parameters of the antioxidative system. It should be emphasized that disturbances in the oxidative/antioxidative balance were observed in the period of clinical exacerbation of asthma, as well as in the period of clinical remission. This fact may constitute further evidence for a prolonged existence of minimal inflammation in the respiratory tract in bronchial asthma. PMID- 11852806 TI - [GM-CSF in the culture supernatants of neutrophils and serum levels in patients with inflammation and squamous cell carcinoma of oral cavity]. AB - The aim of this study was to comparison GM-CSF concentrations in the culture supernatants of neutrophils (unstimulated and LPS-stimulated) and the serum of patients with inflammation and squamous cell carcinoma of oral cavity. PMN from both patient groups exhibited decreased the ability to release of GM-CSF. PMN from patients with inflammation secreted a lower concentrations of GM-CSF than those from cancer patients. PMN from patients with inflammation were more reactivity after LPS-stimulation in comparison with cells from cancer group. GM CSF serum levels in patients with inflammation and cancer were nonsignificantly higher in comparison with control. There was no correlation between GM-CSF in the culture supernatants of PMN and the serum levels in patient groups. The decreased GM-CSF production may lead to unfavourable effects for immune response of patients with inflammation as well as patients with cancer disease. PMID- 11852805 TI - [Tuberculous pleurisy--still difficult diagnostic problem]. AB - In years 1989-2000 in Lung Diseases Hospital in Lodz tuberculous pleurisy TP was diagnosed in 271 patients; in 130 (48%) cases this diagnosis was confirmed by microbiological and histopathological examinations. Presence of tubercle bacilli in pleural fluid was discovered by smear and culture only in 13 cases, only by culture in 95 and in Bactec method in 7. During this period in 135 patients with pleural effusion of unknown etiology biopsy of the pleura (with Abrams needle) was performed and specific granuloma was found in 17 (13%) persons. In 15 (12%) patients it was the only method establishing diagnosis of TP (because in 2 remaining patients tubercle bacilli in pleural fluid were also found). Assessment of clinical symptoms indicates that onset of TP was insidious and course of the disease was subacute in about 30% of patients, manifested mainly by: fatigue, general weakness, night sweats. Radiological estimation of patients with TP disclosed (besides pleural effusion) lung infiltrations in 38 (28%) cases, including 13 (10%) with cavitation. Tuberculin induration with diameter 0 mm was found in 32% of patients with TP and with greater than 10 mm in 29% of patients. Because of low sensitivity of microbiological examinations of pleural fluid for tubercle bacilli biopsy of pleura should be method of choice in the diagnosis of TP in selected cases. PMID- 11852807 TI - [Receptors of selected cytokines and angiokine bFGF in patients with colorectal cancer (a preliminary study)]. AB - The aim of the study was to examine the frequency of increased serum levels of the soluble receptors TNF and IL-2 and angiokine bFGF in colorectal cancer patients. Also correlation between their concentrations and stage of the tumor was made. The study was done on group consisted of 30 diagnosed colorectal cancer patients, with different location and stage of the tumor. The used classification of stage of the tumor was described by Dukes. The results were compared with control group consisted of 10 healthy persons. The examined factors were assayed by ELISA method (R&D Systems Minneapolis). In colorectal cancer group the serum levels of sTNFRI were increased 1.8 times, sTNFRII 1.4 times, sIL-2R 2.2 times and bFGF 5.3 times in comparison with control group. The serum levels of sTNFRI and sTNFRII showed increased tendency in stage D of colorectal cancer. The serum levels of sIL-2R were the highest in stage D. The serum levels of bFGF showed increased tendency in stage A and B and correlated with stage D of the tumor. This results permit for further study on usefulness of sTNFRI, sTNFRII, sIL-2R and bFGF as a markers for colorectal cancer in clinical use. PMID- 11852808 TI - [Involvement of WT1 gene expression in regulation of P53 and MDM2 proteins function in acute lymphoblastic leukemia]. AB - Using sensitive techniques such as reverse transcription polymerase chain reaction (RT-PCR) the expression of WT1 gene in acute lymphoblastic leukemias (ALL) is indicated. High level of mRNA WT1 was only observed in ALL cases with leukemic cells characterized by P53- and MDM2-positive staining in cytometric analysis. The overexpression of P53 protein has not been induced by P53 gene abnormalities and MDM2 protein synthesis was independent from respective gene amplification. The data suggest that WT1 may play a distinct role in the pathophysiology of acute leukemias. It can regulate the function of the main oncoprotein network factors--P53 and MDM2 proteins. There was concluded that the most important mechanism of tissue P53-immunopositivity was connected with the P53 interactions with other oncoproteins, especially with MDM2 and WT1. They have caused different effects in particular cases and several phenotypes of leukemic cells were described. However, the negative tissue staining with anti-P53 monoclonal antibodies can not be evidence of the proper P53 protein function. The immunohistochemical estimations of the P53 level in the cells are insufficient for diagnostic and clinical evaluations. Molecular analyses of P53 and MDM2 genes, as well the WT1 gene transcription, are necessary for the proper characterisation of functional and structural status of P53. PMID- 11852809 TI - [Side effects during dobutamine stress echocardiography in patients with aortic stenosis]. AB - The purpose of the study was to assess the safety of the dobutamine stress echocardiography (DASE) in patients with aortic stenosis (AS). 161 patients (mean age 59 +/- 13 years) with AS were prospectively studied with DASE. There were 58 female and 103 male. Dobutamine was given in stepwise increasing doses from 5 to 40 ug/kg/min. Mean maximal dose achieved was 31.4 ug/kg/min. The test was positive in 40 (24.8%) patients. Significant coronary artery disease was present in 60 (37.3%) patients. DASE resulted in significant increase in transvalvular mean gradient from 29.3 +/- 12.5 mmHg at rest to 46.3 +/- 19.3 mmHg at peak dose. There was no significant increase in valve area. There were no death, myocardial infarction or episodes of sustained ventricular tachycardia as a result of DASE. The test was terminated when following conditions were revealed: target heart rate (39.1%), left ventricular asynergy (25.5%), maximal established dose achieved (8.1%), side effects (27.3%). The most common side effects with the need of test cessation were arrhythmias (9.9%) and hypotension (9.9%). The most side effects were usually well tolerated without need of medical treatment. We conclude that DASE may be safely performed in patients with AS. Side effects are more common than in patients with coronary disease, but are usually well tolerated without need of medical treatment. PMID- 11852810 TI - [Antibodies to tissue transglutaminase as marker of gluten-free diet maintenance in patients with coeliac disease]. AB - The aim of the study was to assess the compatibility of IgA antiendomysium antibodies (IgAEmA) and IgA tissue transglutaminase antibodies (IgAtTG) in treated patients with coeliac disease and to estimate the value of the IgAtTG ELISA as a marker of gluten-free diet maintenance. The study included 71 children and young adults (46 F, 25 M) aged from 6 years to 27 years, on the gluten-free diet (in some cases unrestricted) for at least 4 years because of coeliac disease diagnosed according to ESPGHAN criteria. Serum samples of all patients were examined simultaneously for IgAEmA titer by indirect immunofluorescence and IgAtTG level by ELISA test. IgAEmA antibodies were present in 26 patients (36.6%). IgAtTG-ELISA were positive in 23 patients (32.4%), equivocal--in 8 patients (11.3%) and negative--in 40 patients (53.6%). The serological tests were compatible in 58 patients (81.7%). If we consider equivocal results of IgAtTG test to be negative, IgAEmA and IgAtTG tests will be compatible in 64 cases (90.1%). Tissue transglutaminase antibodies ELISA provide a sensitive test of the gluten-free diet maintenance in patients with coeliac disease. PMID- 11852811 TI - [Plasminogen activators (t-PA and u-PA) and other fibrinolysis parameters in patients with atherosclerosis obliterans and diabetic macroangiopathy]. AB - The aim of the study was to evaluate plasminogen activators in compare fibrinolysis parameters in patients with atherosclerosis obliterans (AO) and diabetic macroangiopathy (DM) of lover extremities. The study group consist of 40 patients with AO (mean age 59 years) and 40 with DM (mean age 61 years). All the patients were diagnosed as the II stage in Fontaine scale. Ischemia lover extremities were: aorta-hip, femoral-popliteal and peripheral dependent closure. The control group consist of 30 healthy volunteers (mean age 51 years). Concentration of: tissue plasminogen activator antigen (t-PA:Ag), urokinase plasminogen activator antigen (u-PA:Ag), plasminogen activator inhibitor type 1 antigen (PAI-1:Ag) (ELISA), PAI-1 activity (PAI-1 act.), euglobulin lysis time (ELT) (Kowarzyk-Buluk method), fibrinogen, fibrin degradation products (FDP) (Merskey method) in blood plasma were evaluated. The results shows statistically higher concentrations of: t-PA:Ag, PAI-1:Ag, fibrinogen, and lower concentrations of u-PA:Ag and elongated ELT in blood plasma patients with AO and DM in compare with healthy volunteers. Comparing analysed parameters between patient groups shows higher concentration FDP in patients with AO than patients with DM. CONCLUSIONS: 1) Although elongated ELT in both groups it was shown evidence of the fibrinolysis activation in the shape of elevated FDP concentration. It was probably compensation for hypercoagulability. 2) Elevated t-PA:Ag concentration in patients with AO and DM is an evidence of the higher releasing of the endothelium in AO and DM. PMID- 11852813 TI - [Etiological factors of urinary tract infections in children]. AB - Etiological factors of urinary tract infection in children have been studied. The Escherichia coli, Klebsiella sp., Enterococcus sp. and Enterobacter sp. were the most often isolated bacteria from infant and baby urine. In this age group of patients uromycoses were also affirmed. From the urine of older children (1-18 years old) mainly Escherichia coli, Enterococcus sp., Proteus sp. and Morganella morganii have been raised. Uropathogens' occurrence was analysed by a statistical method in the relation to age and sex of the children. The girls in the age of 1 to 18 years were more open than boys to urinary tract infections caused by Escherichia coli and less open to infections caused by Morganella morganii and Enterococcus sp. PMID- 11852812 TI - [Laser's biostimulation in healing or crural ulcerations]. AB - The objective of this paper was to evaluate effect of laser's biostimulation on the process of healing of crural ulcerations. Three comparative groups of patients, A, B and C, were made at random from the patients with venous crural ulcerations. The group A consisted of 17, the group B 15, the group C 17 patients. The patients in all comparative groups were treated pharmacologically and got compress therapy. Ulcerations at patients in group A were additionally irradiated by light of biostimulation's laser (810 nm) in this way that every time ulcerations got dose of energy 4 J/cm2. The patient's in-group B additionally got blind trial (with placebo in the form of quasi-laserotherapy). The evaluated factors were to estimate how laser's biostimulation causes any changes of the size of the ulcers and of the volume of tissue defect. The speed of changes of size and volume of tissue defect per week was calculated. After the treatment there was statistically significant decrease of size of ulcers in all comparative groups while there was no statistically significant difference between the groups observed. After the treatment there was statistically significant decrease of volume of ulcers only in groups A and C but there was no statistically significant difference between the groups observed. PMID- 11852815 TI - [Neoplastic skin infiltration in chronic myelomonocytic leukemia]. AB - Chronic myelomonocytic leukemia has been associated with various nonspecific cutaneous manifestations. Rarely has the leucaemia been reported to directly affect the skin. We present a case of 53-year old man with CMML who showed neoplastic cutaneous lesions. The occurrence of these lesions may be the presenting feature of the disease or may herald its progression to acute leukemia. Early diagnosis have therapeutic and prognostic significance. PMID- 11852814 TI - [Appendico-ileo-caecal intussusception as a cause of mechanical ileus]. AB - The authors present a case of 34 years old male operated on for mechanical ileus caused by intussusception of unchanged appendix, cecum and terminal part of ileum. The paper presents possible mechanism of intussusception, diagnostic difficulties determined by non-characteristic symptoms and imaging examination results. PMID- 11852816 TI - [Sarcoidosis on non-typical localization--case report]. AB - We are presenting a 76-year-old women with abdominal pain, constipation and lost of weight. On basis of performed tests we diagnosed sarcoidosis of peripheral and retroperitoneal lymph nodes. PMID- 11852817 TI - [Giant tonsillolith simulating peritonsillar abscess]. AB - The authors describe a tonsillolith detected by a 70-year-old man with odynophagy and a history of often tonsillar infections. A diagnosis of peritonsillar abscess was first made. The large concrement was yellowish-gray, measured 41 x 21 x 19 mm, which was one of the largest reported case in the world. PMID- 11852818 TI - [Proinflammatory cytokines in humoral immune response]. AB - Proinflammatory cytokines secreted by cells involved in inflammatory process play a key role in the regulation of immune responses, both cellular and humoral immune response. Through their influence on T-helper lymphocytes and antigen presenting cells including lymphocytes B, they induce secretion of secondary cytokines and, in cooperation with them, promote activation and proliferation of lymphocytes B and production of variety of classes of immunoglobulins. Cytokines interact with their specific receptors which usually occur both in membrane bound and soluble form. The purpose of this article was to evaluate the relationships between primary cytokines such as interleukin-1 (IL-1) and tumor necrosis factor alpha (TNF-alpha) and other cytokines, lymphocytes T-helper and lymphocytes B in the light of current opinions regarding this matter. PMID- 11852819 TI - [The role of cyclooxygenase and prostaglandins in the pathogenesis of rheumatoid arthritis]. AB - Rheumatoid arthritis (RA) is a systemic inflammatory disease with polyarticular synovitis leading to formation of rheumatoid pannus and subsequent erosion of articular cartilage and bone. Prostaglandins (PGs)--a group of arachidonic acid metabolites found at elevated levels in synovial fluid and synovial membrane are considered to play a pivotal role in development of vasodilatation, fluid extravasation and pain in synovial tissues. Moreover, there is increasing evidence that PGs (especially prostaglandin E2) are mediators involved in complex interactions leading to development of erosions of articular cartilage and juxta articular bone. Cyclooxygenase is an enzyme playing crucial role in PGs production. It is known that two forms of cyclooxygenase exist: cyclooxygenase-1 (COX-1) playing house-keeping functions and cyclooxygenase-2 (COX-2) involved in inflammatory responses. Synovial tissues from patients with RA are shown to contain COX-2 and to a less extent COX-1. COX-2 expression in rheumatoid synovium is induced by proinflammatory cytokines, mainly IL-1, while corticosteroids are capable of inhibiting COX-2 expression. The understanding of crucial role of COX 2 in synovial inflammation led to development of new group of anti-inflammatory agents--selective COX-2 inhibitors, that inhibit specifically COX-2, providing effective anti-inflammatory action without the side effects associated with inhibition of COX-1. In the context of widespread use of selective COX-2 inhibitors hypothetical role of COX-1 in RA pathology should be elucidated. PMID- 11852820 TI - [The role of endothelins in human cardiovascular disease]. AB - Endothelin-1 was first identified by Yanagisawa in 1988 and shown to be a potent and sustained vasoconstrictor and pressure peptide. Endothelial cells line all blood vessels and are capable of generating endothelin-1; receptors for the endothelins are widely expressed, particularly in tissues involved in cardiovascular regulation, including the heart, blood vessels, kidney and brain. Endothelin-1 has potent vasoconstrictor properties and the coronary, renal and cerebral blood vessels appear particularly sensitive. Moreover, endothelin-1 has activity as co-mitogen, interacts with the sympathetic nervous and renin angiotensin system. These properties indicate a likelihood that the endothelin system is of functional importance in human cardiovascular physiology and may play a role in the pathophysiology of cardiovascular disease. It is possible that endothelin antagonists might be effective in the treatment of diseases associated with intermittent or sustained vasoconstriction. PMID- 11852821 TI - [Immunotropic effects of electromagnetic fields in the range of radio- and microwave frequencies]. AB - On the grounds of reviewed literature and the results of own experiments, the authors present current views on the possible immunotropic influence of low energy electromagnetic fields, in the range of radio- and microwave frequencies. They conclude, that a more systematic and multidisciplinary investigations should be undertaken, comprising the wide spectrum of immune homeostatic tasks, including defensive, immunoregulatory and pro-regenerative capabilities of immune system exposed to rapid environmental spread of different electromagnetic emitters. PMID- 11852822 TI - [Methotrexate--mechanism of action and application]. AB - The paper present modern views on turnover, mechanism of action and application of methotrexate. Contraindications and the most frequent side effects observed in certain number of patients treated with this drug are discussed. PMID- 11852823 TI - [Helicobacter pylori--does it only cause gastroduodenal disease?]. AB - Helicobacter pylori is a human pathogen that can be found all over the world. It is responsible for the following diseases of gastrointestinal tube: gastritis, gastric ulcer, duodenal ulcer, gastric cancer, gastric lymphomas, Menetier disease. Some research has been done recently trying to identify the connection between H. pylori infection and idiopathic Parkinson's Disease morbidity. Some of them show that people with this neurological disease are more likely to have ulcers and also seropositivity in the direction of H. pylori. The direct influence of H. pylori infection on Parkinson Disease is not known but the following relations are suggested: H. pylori may produce toxins that damage substantia nigra in brain; possible cross reaction of h. pylori antibodies with dopaminergic neurons; indirect influence of antacids containing aluminium used to alleviate the symptoms of ulcers. Investigations of the reasons for idiopathic parkinson disease draw attention to the influence of food factors. Some researches show that there is a relation between the frequency of eating certain foods and the parkinson disease morbidity. We have numerous techniques that allow us to diagnose h. pylori infection. Those techniques have different sensitivity, accuracy, invasiveness and costs, which determines their usefulness in clinical diagnostics. Approach to eradication of bacteria is still discussed because H. pylori infection doesn't always lead to health problems. Polish Working Group on Helicobacter pylori, called by the National Consultant's Team on Gastroenterology explained clearly when eradication is advisable and when it can be waived. PMID- 11852824 TI - [Mental disorders in Lyme disease]. AB - From the early 90-ties there is a growing number of patients suffering from Lyme Disease all over the world, including Poland. Lyme Disease is the disorder connecting physicians of various specialties. The authors reviewed literature on mental disorders in Lyme Disease during different stages and in different types of illness. Mental disorders are part of clinical picture of the acute stage of Lyme Disease, and could also be its sequel. The most commonly found mental disorders are: encephalopathy, other cognitive disorders, mood disorders (depression), anxiety disorders and less often: psychotic disorders and eating disorders (anorexia nervosa). PMID- 11852825 TI - Introduction to qualitative research. AB - The purpose of this article is to introduce practicing nurses to qualitative research. Qualitative research terms are defined, philosophy of science is briefly discussed, and several types of qualitative research studies are described. A hermeneutic phenomenological study of infant colic is described as an example of qualitative research useful in gastroenterology nursing. Finally, suggested criteria for evaluating a qualitative study are introduced. PMID- 11852826 TI - Endoluminal gastroplication. A new therapeutic endoscopic procedure for gastroesophageal reflux disease. AB - It is estimated that more than 15 million Americans suffer daily from gastroesophageal reflux disease (GERD) (De Vault, 1995). The spectrum of symptoms for GERD ranges from simple heartburn and regurgitation to persistent esophageal tissue damage with subsequent development of serious complications. The frequent occurrence and intensity of GERD symptoms can severely impact a person's quality of life (Behar, 1990). Until recently, treatment options included a lifetime of drug therapy, disagreeable daily lifestyle changes, and the possibility of invasive surgery. In 2000, the Food and Drug Administration cleared a new endoscopic tool called the EndoCinch, a device that enables endoscopic endoluminal gastroplication as an outpatient procedure for the treatment of GERD. Endoscopic endoluminal gastroplication has been shown to be a safe and effective therapeutic outpatient procedure offering sustainable clinical benefits and cost savings to patients suffering with GERD (Patel, 2001; Raijman, 2001; Weiland, 2001). The purpose of this article is to present an overview of GERD with a particular focus on management of the disease using this new procedure. PMID- 11852827 TI - Case management in the GI Clinic. AB - Nursing case management has been in practice since the 1800s when it was first created to coordinate and improve healthcare delivery to the poor and immigrant populations in the United States. In the 1950s and again in 1972, the government mandated programs that provided a community-based continuum of care for the mentally ill and elderly populations. Historically a community service, nursing case management moved into the acute care setting in the 1980s when Medicare implemented its prospective payment system organized as diagnosis-related groups, thus prompting hospitals to use nurse case managers to coordinate care and ultimately reduce costs. PMID- 11852828 TI - M2A capsule endoscopy. A breakthrough diagnostic tool for small intestine imaging. AB - It is estimated that 19 million people in the United States may suffer from diseases related to the small intestine, including obscure bleeding, irritable bowel syndrome, Crohns disease, chronic diarrhea, and cancer. Current diagnostic methods, including push enteroscopy and radiology, produce historically low diagnostic yield. Early studies show that the M2A Capsule Endoscope effectively visualizes the entire small bowel and demonstrates a 71% superior diagnostic yield when compared to push enteroscopy according to clinical trials reviewed by the FDA. In addition to potentially higher diagnostic yields, it provides a noninvasive diagnostic alternative where none previously existed. The M2A Capsule Endoscope received FDA clearance in August 2001 and is currently in use at major medical centers throughout the United States and Europe. PMID- 11852829 TI - Women's colonic digestive health. AB - Digestive problems in women are common and sometimes debilitating. Twenty percent of women suffer from irritable bowel syndrome, 20% have constipation, and all American women will need screening for colon cancer, the number two cause of cancer death in women. This article reviews management of these disorders as well as lower gastrointestinal symptoms associated with menses, hysterectomy, fecal incontinence, and rectal bleeding. PMID- 11852830 TI - Looking at research. PMID- 11852831 TI - Chronic hepatitis C. PMID- 11852832 TI - Forest fires, air pollution, and mortality in southeast Asia. AB - I assess the population health effects in Malaysia of air pollution from a widespread series of fires that occurred in Indonesia between April and November of 1997. I describe how the fires occurred and why the associated air pollution was so widespread and long lasting. The main objective is to uncover any mortality effects and to assess how large and important they were. I also investigate whether the mortality effects were persistent or whether they represented a short-term, mortality-harvesting effect. The results show that the smoke haze from the fires had a deleterious effect on the health of the population in Malaysia. PMID- 11852833 TI - Changing chronic disease rates and long-term declines in functional limitation among older men. AB - Functional limitation (difficulty walking, difficulty bending, paralysis, blindness in at least one eye, or deafness in at least one ear) in the United States fell at an average annual rate of 0.6% among men aged 50 to 74 from the early twentieth century to the early 1990s. Twenty-four percent of this decline is attributable to reductions in the debilitating effects of chronic conditions, 37% is attributable to reduced rates of chronic diseases, and the remainder is unexplained. The findings have implications for theories of the impact of declining mortality rates on the health of elderly people. PMID- 11852834 TI - A dynamic analysis of the effect of child care costs on the work decisions of low income mothers with infants. AB - Child care costs reduce the net benefit of working and consequently influence mothers' decisions to work. They affect the employment of low-income mothers in particular because they represent a larger portion of these mothers' earnings. I used a hazard framework to examine a mother's decisions about work and hours of work after childbirth. I focused on low-income mothers with infants because they are the ones who may be most affected by child care costs. The results showed that child care costs are a barrier to work that is larger for low-income mothers than for non-low-income mothers. Further, child care costs have large negative effects on hours of work. PMID- 11852835 TI - Child care subsidies and the employment of welfare recipients. AB - Changing patterns of maternal employment, coupled with stronger work requirements for welfare recipients, are increasing the demand for child care. For many families, the cost of child care creates a financial burden; for mothers with low incomes and those who are former welfare recipients, these costs may be an insurmountable barrier to employment or economic self-sufficiency. Despite increased public spending in this area, the receipt of any child care subsidy appears to be a relatively rare and uncertain event. In this study, we use data from a sample of low-income single mothers (current and recent welfare recipients in California) to estimate the probability of their receiving child care subsidies and the effect of this probability on labor market activity. PMID- 11852836 TI - The effect of stepchildren on childbearing intentions and births. AB - This article reports on a study of the effect of stepchildren (children from previous unions) on couples' fertility intentions and childbearing behavior using longitudinal data from the National Survey of Families and Households. The results indicated that stepchildren negatively affect childbearing intentions and childbearing risks. Intentions to have a child are weakened by one's own previous biological children and the previous biological children of one's current spouse or partner. This effect varies by the parenting configuration of the couple and gender of the respondent. Among couples with stepchildren, intentions remain high until each partner has had a biological child. Unlike women, men's previous biological children do not affect their intentions of having a child. Stepchildren exert a weak negative effect on couples' childbearing risks, and this effect is mediated by the couples' childbearing intentions. The findings suggest that stepchildren should be incorporated into future models of fertility. PMID- 11852837 TI - On the validity of retrospective assessments of pregnancy intention. AB - Information on pregnancy intention is often gathered retrospectively (after the birth of a child). This article investigates whether the retrospective assessment of pregnancy intention leads to biased estimates of the extent or consequences of unintended fertility. Comparisons are made between pregnancy intentions ascertained during pregnancy and after birth using data from the National Longitudinal Survey of Youth. To address the bias caused by selective recognition or acknowledgment of pregnancy, we used the longitudinal feature of the data to determine actual pregnancy status at the time of interviews, which, in turn, was used as an instrumental variable for the retrospective (versus prospective) reporting of pregnancy intention. After correction for selective pregnancy recognition, we found no evidence that the retrospective assessment of pregnancy intention produces misleading estimates of either the number or the consequences of unintended births. This finding is supported by additional analyses of a small subsample of women for whom information on pregnancy intention was collected both during pregnancy and after birth. PMID- 11852838 TI - AIDS and the elderly of Thailand: projecting familial impacts. AB - We apply aggregate demographic analysis and computer microsimulation to project the number of older Thais who will lose children to AIDS during their own lifetimes and to assess their involvement with ill children through caregiving and coresidence. Parental bereavements from AIDS are predicted to peak at around 80,000 per year between 2003 and 2007. Despite an HIV prevalence of only 2%, 13% of Thais who were over age 50 as of 1995 are likely to experience the loss of at least one adult child to AIDS, and 12% of them will lose multiple children. The chance of losing an adult child during one's lifetime will be 70% higher than if there were no AIDS epidemic. PMID- 11852839 TI - The effects of kin on child mortality in rural Gambia. AB - We analyzed data that were collected continuously between 1950 and 1974 from a rural area of the Gambia to determine the effects of kin on child mortality. Multilevel event-history models were used to demonstrate that having a living mother, maternal grandmother, or elder sisters had a significant positive effect on the survival probabilities of children, whereas having a living father, paternal grandmother, grandfather, or elder brothers had no effect. The mother's remarriage to a new husband had a detrimental effect on child survival, but there was little difference in the mortality rates of children who were born to monogamous or polygynous fathers. The implications of these results for understanding the evolution of human life-history are discussed. PMID- 11852840 TI - Population momentum for gradual demographic transitions: an alternative approach. AB - In this article, I derive a simple formula for approximating the ultimate size of a population that undergoes a gradual transition to replacement fertility. I model the fertility transition by specifying a linear frontier on the Lexis surface across which a change in fertility is instantaneous. Gradual transitions result from variations in the slope of this frontier. This framework can be used to reproduce and understand previous studies of population momentum and gradual transitions. PMID- 11852841 TI - Simultaneity in the use of maternal-child health care and contraceptives: evidence from developing countries. AB - This study examined the relationship between the use of maternal-child health (MCH) care and the use of contraceptives. The high correlation between the two may be due to the independent effect of one on the other or to an association of both with the same or similar background factors. We used structural equation models to examine the relationship between these two interventions. The data were derived from six Demographic and Health Surveys: Zimbabwe from Sub-Saharan Africa, Thailand from Asia, Egypt and Tunisia from North Africa, and Guatemala and Colombia from Latin America. The results show that in all six countries, the use of contraceptives and MCH care are significantly associated, independent of intervening factors; this finding suggests that families develop a joint demand for better-quality health and limited family size and translate these demands into action by using health services for mothers and for children and by voluntarily regulating fertility. PMID- 11852842 TI - Educational stratification by ethnicity in China: enrollment and attainment in the early reform years. AB - Using evidence about educational disparities, this article demonstrates the need for attention to minority populations in studies of social stratification in China. Analyses of data from a 1992 survey of children demonstrate substantial ethnic differences in enrollment among rural 7- to 14 year olds, with rates for ethnic Chinese boys roughly double those for girls from certain ethnic groups. Multivariate analyses indicate that the ethnic gap can be attributed, in part, to compositional differences in geographic location of residence and socioeconomic background. There is no general tendency of a greater gender gap for minorities than for the ethnic Chinese, but significant differences in the gender gap emerge across individual ethnic groups. Together with evidence from census data showing that ethnic disparities in junior high school transitions increased between 1982 and 1990, these results stress the continuing significance of ethnicity as a fundamental factor that conditions status attainment opportunities in China. PMID- 11852843 TI - Iowa Priority Prescription Savings program provides relief for seniors. PMID- 11852844 TI - Competition in health insurance--is there such a thing in Iowa? PMID- 11852845 TI - The problem is not just Medicare. PMID- 11852846 TI - Abstracts of the Annual Symposium of the Czech Society of Histochemistry and Cytochemistry. Brno, Czech Republic, 1-2 December 2000. PMID- 11852847 TI - Abstracts of the 13th National Meeting of the Italian Society for Pediatric Endocrinology and Diabetology (SIEDP). Trieste, 10-13 October 2001. PMID- 11852848 TI - [Congress of the French Society of Sleep Research. 8-10 November 2000, Montpellier, France. Abstracts]. PMID- 11852849 TI - Proceedings of the Conference on Applied Research in Glaucoma and Neuroprotection. September 29-October 1, 2000. Cannes, France. PMID- 11852850 TI - Vestibular and Equilibrium Research: Basic and Clinical Implications. Proceedings of the 21st Barany Society Meeting. Uppsala, Sweden, 4-7 June 2000. PMID- 11852851 TI - Proceedings of the workshop "Exploitation of the Functional Potential of Legume Proteins in the Production of Functional Ingredients for Food and Materials for Non-food Application". 7-8 December 2000. Kleinmachnow, Germany. PMID- 11852852 TI - West Syndrome and other Infantile Epileptic Encephalopathies. Proceedings of an International Symposium. Tokyo, Japan, February 9-11, 2001. PMID- 11852853 TI - Worldwide Endeavor for Epidemiology and Prevention of Cardiovascular Disease. Rome, Italy, 6-7 September 1999. Proceedings of a meeting. PMID- 11852854 TI - Psychological interviewing techniques. PMID- 11852855 TI - An HR oasis. PMID- 11852856 TI - Inaugural Address. 1934. PMID- 11852857 TI - Pharmacology. Paper alert. PMID- 11852858 TI - British Society for Clinical Cytology 40th annual meeting. 9-11 September 2001, Harrogate, United Kingdom. Abstracts. PMID- 11852859 TI - Abstracts of the 23rd annual meeting of the Japanese Society of Biological Psychiatry. 11-13 April 2001. Nagasaki, Japan. PMID- 11852860 TI - [Physiopathology and management of the aged--chronic coughs of aged patients]. PMID- 11852861 TI - Primary epithelioid sarcoma of the dura: case report. AB - OBJECTIVE AND IMPORTANCE: Epithelioid sarcomas are rare mesenchymal neoplasms that occur most often in the extremities of young adults. Despite isolated reports of epithelioid sarcomas arising in the head and neck region, these lesions have not been described previously, to our knowledge, in the central nervous system. CLINICAL PRESENTATION: We present the case of an 18-year-old woman with a unique dural sarcoma that arose in the right frontotemporal region. As visualized on magnetic resonance imaging studies, the 4.5-cm tumor focally traversed the cranium to penetrate the galea, the temporal muscle, and subcutaneous tissue. No brain invasion was noted. INTERVENTION: Despite gross total removal and postoperative radiotherapy (59 Gy), a large recurrence was noted 5 months after surgery. Histologically, the partly necrotic tumor consisted of epithelioid and spindle cells showing widespread vimentin and variable cytokeratin as well as epithelial membrane antigen immunoreactivity. Ultrastructurally, the cohesive cells featured various organelles, intermediate filaments, junctions, and filopodia-containing intercellular spaces. CONCLUSION: With the inclusion of epithelioid sarcoma, the spectrum of central nervous system sarcomas continues to expand. PMID- 11852862 TI - Ethical and religious directives for Catholic health care services. PMID- 11852863 TI - Issues to consider when selling your practice. PMID- 11852864 TI - [Central Venous Access for Hemodialysis: Trends and Perspectives. Montpellier, France, 1-2 February 2001. Proceedings]. PMID- 11852865 TI - Behold, a shining light. PMID- 11852866 TI - Spotlight on Annie Bryant, CNA. Interview by Suzanne Bilyeu. PMID- 11852867 TI - SNFs on an upswing. PMID- 11852868 TI - Castleman's disease: unusual case of inflammatory spinal dorsal epidural mass: case report. AB - OBJECTIVE AND IMPORTANCE: Castleman's disease (giant lymph node hyperplasia) is a rare, heterogeneous, lymphoproliferative disorder of unknown cause and pathogenesis. Most cases occur as mediastinal masses, although extrathoracic involvement, including nodal and extranodal locations, has been reported. The localized variants (solitary lesions) respond well to surgical excision. CLINICAL PRESENTATION: We report the case of a 34-year-old man with headache, intermittent febrile sense, progressive weakness of the legs, and urinary incontinence. Magnetic resonance imaging disclosed an enhancing dorsal extradural mass with impingement on the spinal cord at the T2-T3 level. Other abnormal laboratory findings were increased erythrocyte sedimentation rate and mild leukocytosis. TECHNIQUE: The mass lesion was removed; the histopathological findings included diffuse lymphoid cell infiltration and vascular proliferation, which are compatible with Castleman's disease. CONCLUSION: Although the dorsal epidural site of this case is very unusual, Castleman's disease was considered the most appropriate diagnosis on the basis of the associated systemic findings. This patient with dorsal epidural Castleman's disease may be the first reported case in the literature. PMID- 11852869 TI - An evaluation of the Model C gamma knife with automatic patient positioning. PMID- 11852870 TI - National HIV Prevention conference, Atlanta. PMID- 11852871 TI - Board approves JCAHO corporate priorities and updated strategic plan. PMID- 11852872 TI - Resident safety and error reduction standards approved for long term care. PMID- 11852873 TI - Revised EC standards for design criteria and utilities maintenance. PMID- 11852874 TI - The management of the agitated ICU patient. PMID- 11852876 TI - Infant Nutrition and Pathology. Proceedings of an international symposium. May 1999, Granada, Spain. PMID- 11852875 TI - AIDS. Early warning. PMID- 11852877 TI - Letter from the secretary. PMID- 11852878 TI - The subject of dental anaesthesia. PMID- 11852879 TI - Physical and psychological stress in rats enhances colonic epithelial permeability via peripheral CRH. AB - Stress may be a contributing factor in intestinal inflammatory disease; however, the underlying mechanisms have not been elucidated. We previously reported that acute stress altered jejunal epithelial physiology. In this study, we examined both physical and psychological stress-induced functional changes in colonic mucosa. Colonic mucosal tissue from rats subjected to either 2 hr of cold restraint stress or 1 hr of water-avoidance stress demonstrated altered ionic transport as well as significantly elevated baseline conductance (ionic permeability) and flux of horseradish peroxidase (macromolecular permeability). Intraperitoneal pretreatment with the corticotropin-releasing hormone (CRH) antagonist, a helical CRH(9-41), inhibited the stress-induced abnormalities, while exogenous intraperitoneal administration of CRH, to control rats, mimicked the stress responses and in vitro CRH increased the macromolecular permeability. These results suggest that peripheral CRH mediates stress-induced colonic pathophysiology. We speculate that a stress-induced barrier defect may allow uptake of immunogenic substances into the colonic mucosa, initiating or exacerbating intestinal inflammation. PMID- 11852881 TI - Most patients overdose on topical nasal corticosteroid drops: an accurate delivery device is required. PMID- 11852880 TI - NSAID gastric ulceration: predictive value of gastric pH, mucosal density of polymorphonuclear leukocytes, or levels of IL-8 or nitrite. AB - NSAID use and Helicobacter pylori both cause damage to the gastric mucosa and can cause peptic ulcers. Our aim was to test the relationship between gastric mucosal polymorphonuclear leukocyte (PMN) infiltration and the severity of NSAID-induced gastric injury. H. pylori density, mucosal interleukin-8 (IL-8), and nitrite levels were assessed after receiving placebo and again after receiving 1000 mg of naproxen daily for three days. Histology was graded using a visual analog scale (0-5). IL-8 levels were assayed by ELISA and nitrite levels by Griess reaction. Eleven healthy volunteers with H. pylori infection entered. All had normal appearing gastric mucosa after placebo. Postnaproxen gastric damage included three with none, one with mild, three with moderate, two with severe, and three were very severe mucosal injury (including one with an ulcer >5 mm). There was an inverse correlation between endoscopic score and the pH of the gastric juice post therapy (R = -0.77, P = 0.004). There was no significant change in histologic or biochemical parameters from pretreatment levels. And none of the parameters (eg, PMN density) predicted endoscopic outcome. In conclusion, there was no relation between mucosal PMN density and endoscopic mucosa injury. PMN infiltration, while not predictive, may be a surrogate for an H. pylori infection-related increased risk of NSAID ulcers. PMID- 11852882 TI - A single mutation 202G>A in the human glucose-6-phosphate dehydrogenase gene (G6PD) can cause acute hemolysis by itself. PMID- 11852883 TI - Catheter-related thrombosis in children with hemophilia A: evidence of a multifactorial disease. PMID- 11852884 TI - Hospital-based family caregiver programs: building institutional resources and community ties. PMID- 11852885 TI - State certification of mammography facilities. Final rule. AB - The Food and Drug Administration (FDA) is amending its regulations governing mammography. The amendments implement the "States as Certifiers" (SAC) provisions of the Mammography Quality Standards Act of 1992 (MQSA). These amendments permit FDA to authorize individual States to certify mammography facilities, conduct facility inspections, enforce the MQSA quality standards, and administer other related functions. The amendments establish the standards to be met by States receiving this authority. They also establish procedures for application, approval, evaluation, and withdrawal of approval of States as certification agencies. FDA retains oversight responsibility for the activities of the States to which this authority is given. Mammography facilities certified by those States must continue to meet the quality standards established by FDA for mammography facilities nationwide. PMID- 11852886 TI - Is there a role for implantable cardioverter defibrillators in long QT syndrome? AB - The congenital familial long QT syndrome (LQTS) is characterized by QT interval prolongation on ECG and potentially life-threatening polymorphic ventricular arrhythmias. Antiadrenergic therapy, i.e., beta-adrenoceptor blockade, left cardiac sympathetic denervation, and occasionally pacemaker therapy, sufficiently protects most LQTS patients. Implantable cardioverter defibrillator treatment, with some specific problems and setting requirements in LQTS patients, should at least be considered or implanted in patients with recurrent arrhythmias despite adequate antiadrenergic therapy. Some genetic subtypes, such as LQTS3, may not respond as well (or even adversely) to antiadrenergic therapy and, thus, benefit more from implantable cardioverter defibrillator therapy. PMID- 11852887 TI - Echocardiographic predictors of survival in patients undergoing radiofrequency ablation of postinfarct clinical ventricular tachycardia. AB - INTRODUCTION: The aim of this study was to determine the predictive value of echocardiographic parameters of systolic left ventricular (LV) dysfunction for survival in a group of patients with "mappable" ventricular tachycardia (VT) after myocardial infarction who underwent radiofrequency ablation (RFA) of their clinical VT(s). METHODS AND RESULTS: RFA of at least one inducible, "mappable," and clinical VT was attempted in 61 patients. In total, 63 (79%) of 80 target clinical VTs were ablated successfully, such that clinical VT(s) were noninducible in 49 (80%) of 61 patients. At the last recorded follow-up (range 2 to 98 months; mean 21 +/- 20), nonfatal VT recurrences were observed in 11 (22%) patients; 10 (16%) patients died. On univariate analysis, a higher LV end diastolic volume (LVEDV; P = 0.008) and, by multivariate analysis, applying backward selection of variables, older age (P = 0.03) with a higher LVEDV (P = 0.003) predicted patients more likely to die. When age and LV ejection fraction (LVEF) were excluded, LV end-systolic diameter (LVESD; P = 0.007) was the most significant predictor of survival. CONCLUSION: In our patient population with postinfarct VT who underwent RFA of mappable clinical VT(s), LVEF did not predict survival. In this group of patients with overall low mean LVEF (<35%), older age together with higher LVEDV and LVESD predicted patients who were more prone to die. LV size rather than LVEF correlated with survival. PMID- 11852888 TI - Future of device therapy for arrhythmias. AB - During the last three decades, implantable devices have been developed that are able to treat complex arrhythmias on an individualized basis, with emphasis on rapid restoration of normal cardiac rhythm or creation of the best physiologic alternative. Currently, new areas for device therapy include atrial fibrillation, life-threatening ventricular arrhythmias (with or without heart failure), and out of-hospital sudden death. To improve resuscitation from circulatory arrest, the development and widespread use of dependable devices continuously registering cardiac rhythm and able to localize the victim are needed to shorten the time interval to successful defibrillation. PMID- 11852889 TI - Mortality and incidence of atrial fibrillation in paced patients. AB - This review presents and discusses available data from randomized controlled trials on the prognosis of pacemaker patients, especially the incidences of atrial fibrillation (AF) and death, the impact of pacing mode selection, and the impact of AF on prognosis. The incidence of AF is several times higher in paced patients than in the nonpaced population. The annual incidences of AF and chronic AF are at least 5% and 3%, respectively, after pacemaker implantation. Mean lifetime cumulative incidences of AF and chronic AF can be estimated at approximately 30% to 40% and 20%, respectively. The most important predictors of AF are brady-tachy syndrome, sick sinus syndrome, and selection of VVI(R) pacing mode. The expected lifespan in paced patients is shorter than in the age-matched nonpaced population. One of the factors decreasing lifespan in paced patients most likely is the high incidence and prevalence of AF. In patients with sick sinus syndrome, VVI pacing significantly increases AF and mortality compared with AAI pacing. In a mixed population of patients with bradycardia, DDD(R) pacing causes less often than does VVI(R) pacing. Survival does not differ between these pacing modes within the first 3.5 years after pacemaker implantation. At the present time, AAI(R) should be the preferred pacing mode in patients with sick sinus syndrome, and DDD(R) should be used for other patients without chronic AF for prevention of AF. It is not clear whether prevention of AF will improve survival of paced patients. PMID- 11852890 TI - Outcome studies with device therapy in patients with heart failure. AB - Heart failure is a common debilitating condition for which pharmacologic therapy thus far has provided only partial relief. Despite, and sometimes because of, medical therapy, the overall prognosis remains poor, with high rates of sudden death and death from progressive heart failure. Device-based therapies offer considerable promise for relief of symptoms and for improving prognosis. It is clear that implantable defibrillators should be considered for patients with heart failure who have been resuscitated from ventricular fibrillation or sustained ventricular tachycardia. Several large studies currently are investigating the effects of implantable defibrillators on total mortality in patients with major left ventricular systolic dysfunction but without other risk factors for sudden death. Cardiac resynchronization is a promising new therapy that may relieve the symptoms of heart failure in appropriately selected patients resistant to optimal pharmacologic therapy. Two large trials (CARE-HF and COMPANION) currently are investigating the effects of cardiac resynchronization therapy (CRT) on morbidity and mortality. It is important that those involved in these trials enroll patients quickly and minimize device implantation into patients who have not been assigned this therapy (cross-overs). Overenthusiasm for the benefits that doctors believe devices might bring could destroy the future basis for our clinical practice, denying future generations of patients and the doctors themselves access to what they believe to be effective treatments. PMID- 11852891 TI - High postoperative risk after pneumonectomy in elderly patients with right-sided lung cancer. AB - The present study investigated postoperative mortality (POM), its predictors and relationship with long-term survival in patients who underwent surgery for lung cancer. The 30-day mortality after thoracotomy in 1,830 patients from the Flemish multicentre hospital-based lung cancer registry was analysed according to patient, tumour, treatment and hospital characteristics and compared with 5-yr survival figures for the same patients. Overall POM was 4.4%. In univariate analysis age, extent of surgery and low hospital volume were associated with a higher POM. In multiple regression analysis age, extent of surgery and side of the pneumonectomy proved to be independent predictors of POM. In patients aged >70 yrs who underwent right-sided pneumonectomy POM was 17.8%. Overall, mortality was comparable to published series from referral centres. Age and extent of resection are the main predictors of postoperative mortality in lung-cancer patients. In the operable elderly patient, age alone does not justify denying the survival benefit experienced by resection of lung cancer. The high mortality after right-sided pneumonectomy in elderly patients warrants caution, as the treatment benefit may become marginal. PMID- 11852892 TI - Environmental tobacco smoke and health in the elderly. AB - The aims of this article are to synthesize the evidence on health effects of environmental tobacco smoke (ETS) in the elderly and to discuss questions for future research. Health effects are divided into aetiological and prognostic studies. There is convincing evidence that ETS causes lung cancer and coronary heart disease, both of which are diseases of the elderly. Several cross-sectional studies show increased occurrence of chronic respiratory symptoms and deficits in ventilatory lung function in relation to ETS exposure at home and/or at work. A limited number of studies have found significant relations between ETS exposure and asthma, chronic obstructive pulmonary disease (COPD), pneumococcal infections and stroke in the elderly. Longitudinal studies are needed before any definite conclusions can be made concerning ETS and noncarcinogenic respiratory diseases in the elderly. The potential role of environmental tobacco smoke exposure as a prognostic factor determining development of a pre-existing respiratory or heart disease is an important new area for research. PMID- 11852893 TI - Evaluation of alveolar surfactant aggregates in vitro and in vivo. AB - In acute lung injury, a decrease in surface-active large aggregates and an increase in the less surface-active small surfactant aggregates are observed. The objective of the current study was to determine if the increase in small aggregates interfered with the function of large aggregates, thereby independently contributing to lung dysfunction. Isolated large aggregates, small aggregates, and large aggregate+small aggregate combinations were analysed for in vitro surface activity utilizing a pulsating bubble surfactometer. Subsequently, large aggregates, small aggregates, and large aggregate+ small aggregate combinations were administered to surfactant-deficient, adult Sprague-Dawley rats. Physiological parameters were measured during 1 h of ventilation. After sacrifice, the whole lung lavage was analysed for protein concentration, and surface activity of the recovered large aggregates. The minimum surface tension of the large aggregate+small aggregate preparations (10 mN x m(-1)) was significantly higher than large aggregates alone (1 mN x m(-1)), but lower than small aggregates alone (21 mN x m(-1) ) after 100 pulsations. In vivo, rats receiving large aggregates+small aggregates showed immediate increases in oxygenation, similar to animals given large aggregates, whereas animals given small aggregates and control animals maintained low oxygenation values. In conclusion, small aggregates interfered with large aggregates function in vitro, but this was not observed in vivo in this experimental model. PMID- 11852894 TI - Adjustment to chronic obstructive pulmonary disease: the importance of psychological factors. AB - Research has indicated a weak relationship between the degree of physical problems and quality of life in patients with chronic obstructive pulmonary disease (COPD). The importance of adaptive psychological functioning to maintain optimum quality of life has long been recognized, but there is a lack of empirical evidence concerning the nature of psychological factors involved in adjustment to COPD. Ninety-two males completed questionnaires to determine their coping strategies, levels of self-efficacy of symptom management and social support. Adjustment was measured in terms of depression, anxiety and quality of life. Symptom severity, socioeconomic status, duration of disease and age, which have been demonstrated to be of consequence in COPD, were used as control variables in hierarchical multiple regression analyses. Higher levels of catastrophic withdrawal coping strategies and lower levels of self-efficacy of symptom management were associated with higher levels of depression, anxiety and a reduced quality of life. Higher levels of positive social support were linked to lower levels of depression and anxiety, while higher levels of negative social support were linked to higher levels of depression and anxiety. To maximize quality of life in patients with chronic obstructive pulmonary disease, psychological factors need to be carefully assessed and addressed. PMID- 11852895 TI - Neutrophil degranulation and cell lysis is associated with clinical severity in virus-induced asthma. AB - Acute exacerbations of asthma are frequently caused by viral infections, but the inflammatory mechanisms in virus-induced asthma are poorly understood. The aim of the present study was to determine whether viral infection in acute asthma was associated with increased sputum neutrophil degranulation and increased cellular lysis and whether these changes are related to clinical severity. Adults (n=49) presenting to the emergency department with acute asthma were examined for infection by means of sputum direct-fluorescence antigen detection, sputum culture, and sputum polymerase chain reaction for Mycoplasma, Chlamydia and Legionella pneumophila, and all common respiratory viruses. Subjects infected with one of these agents were classed as having an infective exacerbation. Spirometry and sputum induction were performed on presentation and 4-5 weeks later. Thirty-seven subjects (76%) had virus infection and acute asthma. Those with virus infection had increased sputum neutrophils (p<0.05) and increased neutrophil elastase (p<0.05), this was related to increased elevated sputum lactate dehydrogenase (LDH). Subjects with noninfective asthma had an increase in the proportion of sputum eosinophils. Both groups had elevated sputum eosinophil cationic protein (ECP) concentrations. Higher levels of sputum LDH and ECP were associated with a longer hospital stay. Virus infection and acute asthma is associated with neutrophilic inflammation, cell lysis and more severe clinical disease. PMID- 11852896 TI - Management of tooth surface loss. AB - This part of the series is devoted to tooth surface loss (TSL) not caused by caries or trauma. The management of this form of generalised TSL is included in this series because knowledge of occlusion is needed for both the diagnosis and, when indicated, treatment. There are, however, many other factors involved in the management of generalised TSL other than those associated with 'occlusion'. These will also be discussed. PMID- 11852897 TI - A survey of the use of matrix bands and their decontamination in general dental practice. AB - AIMS: The aims of this study were to determine the pattern of use and re-use of matrix bands in general practice in Scotland, to demonstrate which type of matrix band is most commonly used and to examine infection control measures of relevance to the safe use and re-use of matrix bands. MATERIALS AND METHODS: SUBJECTS: 621 of Scotland's 1,849 general dental practitioners were randomly selected. DATA COLLECTION: A 19-item self-reported questionnaire was mailed in June 1999 with a follow-up mailing sent in August 1999. ANALYSIS: Data analysis involved descriptive statistics and cross-tabulation. Where appropriate, differences between categories were tested for significance by a Chi-square test. RESULTS: A total of 479 questionnaires were returned, representing a response rate of 77%. Reported compliance with routine glove wearing was high (91%). Most dentists (92%) provided training on instrument cleaning and sterilisation for their dental nurses. Ultrasonic baths were used by 59% of practitioners; the remainder soaked or manually scrubbed instruments to remove debris before autoclaving. The Siqveland matrix was the matrix of choice for 96% of respondents. 7% provided a new matrix band for each patient. Most (64%) changed bands only when they were bent or damaged; 29% changed them daily or weekly. Deterrents to use of a new band for each patient were cost (39%) and time (52%). A total of 54% of respondents considered matrix band replacement unnecessary between patients. CONCLUSIONS: The Siqveland matrix band is the most popular among the study group of dental practitioners. Re-use of matrix bands is common. Guidelines for the safe re-use of matrix bands are required. PMID- 11852898 TI - A study of blood contamination of Siqveland matrix bands. AB - AIMS To use a sensitive forensic test to measure blood contamination of used Siqveland matrix bands following routine cleaning and sterilisation procedures in general dental practice. MATERIALS AND METHODS: Sixteen general dental practices in the West of Scotland participated. Details of instrument cleaning procedures were recorded for each practice. A total of 133 Siqveland matrix bands were recovered following cleaning and sterilisation and were examined for residual blood contamination by the Kastle-Meyer test, a well-recognised forensic technique. RESULTS: Ultrasonic baths were used for the cleaning of 62 (47%) bands and retainers and the remainder (53%) were hand scrubbed prior to autoclaving. Overall, 21% of the matrix bands and 19% of the retainers gave a positive Kastle Meyer test, indicative of residual blood contamination, following cleaning and sterilisation. In relation to cleaning method, 34% of hand-scrubbed bands and 32% of hand-scrubbed retainers were positive for residual blood by the Kastle-Meyer test compared with 6% and 3% respectively of ultrasonically cleaned bands and retainers (P < 0.001). CONCLUSIONS: If Siqveland matrix bands are re-processed in the assembled state, then adequate pre-sterilisation cleaning cannot be achieved reliably. Ultrasonic baths are significantly more effective than hand cleaning for these items of equipment. PMID- 11852899 TI - Ethics--the early division of oral health care responsibilities by Act of Parliament. AB - The question of how the profession of dentistry became what it is today, an independent and vigorous one, is an absorbing study. In this analysis of the legislation of the mid-sixteenth century and its effect on modern oral health care delivery, two Acts in particular are notable for their importance in the development of the self-determining practise of dentistry as we now enjoy it in the United Kingdom. The first of these is the 1540 Act uniting the Barbers' and Surgeons' Companies, and the second is one dating from two years later; 'A Bill that Persons, being no common Surgeons, may minister Medicines, notwithstanding the Statute.' Apart from a brief period of 35 years extending from the Dentists Act of 1921 to the Dentists Act of 1956, when the Dental Board of the United Kingdom (which after 1956 became the General Dental Council), was subject to the over-riding control of the General Medical Council, the delivery of oral health care in England has enjoyed an independence which is here identified as having its origin in these Acts 450 years ago in the reign of Henry VIII. PMID- 11852900 TI - Tribal and geographical variations of lupus in the Sultanate of Oman: a hospital based study. AB - Tribal differences are demonstrated in the presentation of lupus in Omani Gulf Arabs (OGA) and Omani Arabs of Persian descent (OAP), both groups resident in and indigenous to the Sultanate of Oman. The OAP have a lower risk of joint complications, skin rash, and dsDNA antibodies than OGA (beta coefficient and 95% confidence interval [CI], -35.6, -16.4, and -2.31 and -38.05 to -33.06, -19.49 to -13.3, and -4.1 to -0.6, respectively). The OGA have higher levels of IgG isotype APL antibodies than OAP, both anticardiolipin (ACA) (t= 1.75, P= 1.04) and anti beta2glycoprotein I (infinitybeta2GPI) (t=3.64, P=0.004) antibodies, while OAP have higher levels of ACAs, IgM isotype (t=2.86, P=0.0024), than OGA. Antiphospholipid (APL) antibodies associate differently with clinical symptoms, both within OAP and between OAP and OGA. Patients from Dakiliyah have a higher risk of joint disease than those from Muscat (relative risk ratio 5.0, 95% CI 1.3 18.3). Differences in symptomatology suggest variations in genetic linkages to human systemic lupus erythematosus. PMID- 11852901 TI - Deficiencies in microbiological quality and labelling of probiotic supplements. PMID- 11852902 TI - Abstracts of the 14th biennial meeting of the International Society for Developmental Neuroscience. 31 January-4 February 2002. Sydney, Australia. PMID- 11852903 TI - Proceedings of the XVIIth World Conference on Health Promotion and Health Education. Paris, France, July 15-20, 2001. PMID- 11852904 TI - Contribution of a medium-sized tropical river to the particulate heavy-metal load for the South Atlantic Ocean. AB - The Paraiba do Sul River is a medium-sized river, 1145 km in length with a drainage basin of 55,400 km2. The riverine fluxes of particulate metals (Cu, Cr, Zn, Mn and Fe) were investigated over 24 months. Particulate matter samples were monthly collected from April 1994 to March 1996. The first 12-month period presented lower rainfall than the second, although both periods presented average precipitation lower than the regional average. The particulate matter flux in the second period (2,042,080 t) was 250% higher than the first period (821,489 t). The same trend was observed for the associated metals, which presented higher fluxes in the second period. This study highlights the strong dependence of the transported mass on the rainfall, and consequently with the river water discharge. The Paraiba do Sul River presents a low contribution to the world oceans, although the local contribution could be considered relevant. PMID- 11852905 TI - Effect of recombinant human bone morphogenetic protein-2/absorbable collagen sponge (rhBMP-2/ACS) on healing in 3-wall intrabony defects in dogs. AB - BACKGROUND: Recombinant human bone morphogenetic protein-2 (rhBMP-2) in an absorbable collagen sponge (ACS) carrier is being evaluated as a candidate therapy for periodontal regeneration. The objective of this study was to evaluate regeneration of alveolar bone and cementum, and associated root resorption and ankylosis following surgical implantation of rhBMP-2/ACS in a canine clinical model. METHODS: Bilateral 3-wall intrabony periodontal defects were surgically induced in the premolar region in the maxilla and mandible in 8 young adult Korean mongrel dogs. The defects in each animal received rhBMP-2/ACS (rhBMP-2 at 0.2 mg/ml, total implant volume/defect approximately 0.1 ml) or buffer/ACS, or served as sham-operated controls. Surgeries were sequenced for each animal to provide postmortem observations following 8- and 24-week healing intervals. Treatment outcomes were evaluated using clinical, radiographic, and histometric parameters. RESULTS: Surgical implantation of rhBMP-2/ACS resulted in accelerated enhanced bone formation in the 3-wall intrabony periodontal defects but in no apparent enhancement of cementum regeneration. rhBMP-2/ACS did not appear to be associated with aberrant healing events such as root resorption and ankylosis under these simulated clinical conditions. CONCLUSIONS: Surgical implantation of rhBMP-2/ACS may be used safely to support regeneration of alveolar bone in intrabony periodontal defects in dogs without aberrant events such as root resorption or ankylosis complicating the regenerative procedure. rhBMP-2/ACS does not appear to have a significant effect on cementum regeneration and formation of a functional periodontal ligament in this model. PMID- 11852906 TI - A comparison of the macrophyte cover and macroinvertebrate fauna at three sites on the River Kennet in the mid 1970s and late 1990s. AB - In 1974-1976, baseline studies were carried out on the flora and macroinvertebrate fauna of the R. Kennet at two sites downstream of Marlborough (Savernake Upper and Lower) and at one site upstream of Hungerford (Littlecote). Simplified maps of each site, showing the cover of macrophytes, were obtained monthly between April 1974 and April/June 1976, and replicated quantitative samples of the macroinvertebrates were collected on the dominant macrophyte and on gravel in June 1974, and also in June and December 1975. As a consequence of two major droughts and increasing concern over water quality in the Upper Kennet in the 1990s, the studies recommenced in the summer of 1997 using the same sites and methodologies. Maps and macroinvertebrate samples were obtained in early July and December 1997 and in June of both 1998 and 1999. At the Savernake sites, mapping in summer 1997 confirmed what had been apparent for some years. That is, macrophyte cover (both Ranunculus and Schoenoplectus) was much lower than in the 1970s. In contrast, the site downstream at Littlecote retained a relatively high cover of Ranunculus, despite the drought. In late autumn 1997, phosphate stripping commenced at Marlborough Sewage Treatment Works, the drought ended and in addition, the spring of 1998 was unusually wet. Ranunculus recolonised both Savernake sites with remarkable speed by summer 1998 and retained this dominant position in 1999. Quantitative samples of macroinvertebrates collected on gravel and the dominant macrophyte at each of the three study sites indicated that there was no evidence of major loss of family richness between the 1970s and 1990s as a result of the low flows or enrichment. However, at Savernake (but not Littlecote) in summer 1997, the macroinvertebrate assemblage was affected by low flows and/or enrichment. This took the form of changes in the abundance of some families, with lentic forms being favoured in relation to some lotic families. Following the end of the drought, many macroinvertebrate families at Savernake showed a rapid response to the new conditions and the assemblages reverted to those expected in a fast-flowing cretaceous chalk stream. Continued monitoring through the next drought is advisable to provide a greater understanding of the interplay between water quality, the discharge regime, habitat quality (including macrophyte growth) and the response of the macroinvertebrate fauna. PMID- 11852907 TI - Phosphorus in soils and field drainage water in the Thame catchment, UK. AB - Field drains were sampled at five farms in a catchment in south-central England. The farms were selected to include the main soil types present in the catchment, stagnogleys and pelosols. The phosphorus content of field drainage water was measured on several occasions from 1999 to 2000, under varying flow conditions. The components measured were soluble reactive phosphorus (SRP), total dissolved phosphorus (TDP) and total phosphorus (TP). SRP concentrations in drainage water were lower than in streams in the catchment which had no apparent point source. This indicated that many streams were receiving unidentified point sources of SRP. Measurements of P in field drainage water samples under high flow conditions showed concentrations of particulate phosphorus and SRP up to 1300 and 300 microg l(-1), respectively, these being associated with high suspended sediment concentrations. Comparison of field drain and soil phosphorus contents at the locations sampled did not provide evidence of an inter-relationship. The equilibrium phosphorus content (EPC0) of surface soil was generally higher than the SRP content of drainage water, at one farm by 1 order of magnitude. The variability in measurements suggested a larger-scale and more focussed survey would be required to characterise catchment-scale phosphorus losses from commercial farms by land use and soil type. PMID- 11852908 TI - Agricultural nutrient inputs to rivers and groundwaters in the UK: policy, environmental management and research needs. AB - Losses of nitrogen (N) and phosphorus (P) in land run-off and drainage from agricultural land can impair river water quality and may pose a potential health hazard. Losses of P are up to an order of magnitude smaller than those of N, but may be more significant with respect to freshwater eutrophication. At the field scale, research suggests that rates of nutrient loss are sensitive to both nutrient and land management, in particular, where nutrient inputs continuously exceed production requirements and where farming methods increase land vulnerability to run-off and erosion. A clear distinction can be made between N and P in the timescales over which inputs of these nutrients are buffered by terrestrial ecosystems against loss, which has implications for control strategies. At the river basin scale, any targets for reducing nutrient loss are best guided by site-specific information on their likely ecological impact, but this information rarely exists for UK rivers affected by eutrophication, and only general guidelines are available. True management of the environment requires integrated approaches which include both N and P taking account of differences in their source areas and delivery mechanisms, the vulnerability of land use and adoption of safe management options in relation to landscape characteristics and the sensitivity of the watercourse along its reach. For P, the identification of vulnerable zones represents a step forward to the management of the river basin in smaller definable units, which can provide a focus for safe management practices. This requires a better understanding of the linkages between nutrient sources, transport and impacts and is considered an urgent research priority. PMID- 11852909 TI - A comparative study of the antipyretic effects of indomethacin and dipyrone in rats. AB - OBJECTIVE: Compare the antipyretic effects of dipyrone and indomethacin. MATERIALS AND METHODS: Fever was induced in rats by i. v. LPS or i. c. v. interleukins (IL), prostaglandins (PG), arachidonic acid (AA), pre-formed pyrogenic factor (PFPF), tumour necrosis factor-alpha (TNF-alpha) or corticotrophin releasing hormone (CRH). Dipyrone and indomethacin were administered i.p., arginine vasopressin V1-receptor antagonist, d(CH2)5 Tyr(Me)AVP, into the ventral septal area. Cyclooxygenase (COX-1/-2) blocking activity was assessed in transfected COS-7 cells. CRH release from isolated hypothalami was determined by ELISA. RESULTS: Indomethacin or dipyrone reduced LPS, IL-1beta, IL-6 or TNF-alpha induced fever and CRH release from rat hypothalamus. Only dipyrone inhibited IL-8, PFPF or PGF2alpha fever. Only indomethacin inhibited fever induced by AA or IL-1beta, plus AA. Neither antipyretic affected fever caused by PGE2 or CRH. d(CH2)5Tyr(Me)AVP only blocked antipyresis induced by indomethacin. Dipyrone at a very high concentration (10 mM) inhibited only COX-1, while indomethacin (0.1 microM) blocked COX-1 and COX-2 in COS-7 cells. CONCLUSION: The antipyretic effect of dipyrone differs from that of indomethacin in that it does not depend on AVP release or inhibition of PG synthesis. PMID- 11852910 TI - Intra-luminal exposure of murine airways to peroxynitrite causes inflammation but not hyperresponsiveness. AB - OBJECTIVE AND DESIGN: There is increasing evidence for the involvement of reactive nitrogen species like peroxynitrite (ONOO-) in airway pathology, for example during allergic airway inflammation. Therefore, the effect of peroxynitrite exposure on airway responsiveness and inflammation was studied. MATERIALS: Male BALB/c mice were treated intra-tracheally with authentic peroxynitrite and the peroxynitrite donor 3-morpholinosydnonimine (SIN-1). Control animals received decomposed solutions of peroxynitrite and SIN- 1. METHODS: Airway inflammation was monitored by bronchoalveolar lavage, three and seven days after administration. Airway responsiveness to methacholine and acetylcholine was measured on day 1, 2, 3 and 7 post administration using whole body plethysmography. RESULTS: Intra-tracheal administration of peroxynitrite 200 microM in 50 microl phosphate buffered saline (PBS) induced a significant increase in macrophages (>35%, p < 0.05) in the airway lumen three days after administration. In contrast, neither intra-tracheal administration of authentic peroxynitrite (up to 5 mM) nor the peroxynitrite donor SIN-1 (1 mM, both intra tracheal and nebulized) changed airway responsiveness to methacholine. Moreover, peroxynitrite (5 mM) did not alter responsiveness to acetylcholine. CONCLUSION: Administration of peroxynitrite directly into the airways of BALB/c mice, induces airway inflammation, but not airway hyperresponsiveness. It is suggested that antioxidants in the epithelial lining fluid and/or the epithelium itself form an efficient barrier, which prevents peroxynitrite from reaching putative targets in the airway interstitium. PMID- 11852911 TI - Neutrophil defensins stimulate the release of cytokines by airway epithelial cells: modulation by dexamethasone. AB - OBJECTIVE AND DESIGN: Neutrophils may contribute to recruiting other cells to sites of inflammation by generating chemotactic signals themselves, or by stimulating other cell types to release chemoattractants such as interleukin-8 (IL-8). Recently, we demonstrated that neutrophil-derived alpha-defensins are able to increase IL-8 expression in airway epithelial cells. In addition, it has previously been reported that neutrophil elastase-induced IL-8 synthesis was insensitive to inhibition by the glucocorticoid dexamethasone. The aim of the present study was to investigate the effect of defensins on the expression of various cytokines in cultured airway epithelial cells and to examine the effect of dexamethasone on defensin-induced cytokine synthesis in these cells. METHODS: Cultures of A549 cells and primary bronchial epithelial cells (PBEC) were stimulated with defensins either alone or in the presence of dexamethasone. Supernatants were analyzed for IL-8, ENA-78, IL-6, MCP-1 and GM-CSF by ELISA. In addition, IL-8 and ENA-78 mRNA was detected by Northern blot analysis. RESULTS: Defensins increased IL-8 expression, ENA-78, MCP-1 and GM-CSF release from A549 cells, whereas in PBEC only IL-8 and IL-6 were increased. Pre-treatment with dexamethasone significantly reduced defensin-induced IL-6, IL-8 and ENA-78 synthesis in airway epithelial cells. In addition, dexamethasone also reduced the neutrophil chemotactic activity in supernatants of these cells. CONCLUSIONS: The results from the present study indicate that defensins differentially induce cytokine secretion by A549 cells and PBEC. Glucocorticoids may interfere with the defensin-induced inflammatory process by reducing defensin-induced cytokine secretion in lung epithelial cells. PMID- 11852912 TI - Technology as a 'major driver' of health care costs: a cointegration analysis of the Newhouse conjecture. AB - Per capita real income on the demand-side and technological change, proxied by total R&D and health R&D spending, on the supply-side are hypothesized as major drivers of per capita real health care expenditure in the US during the 1960-1997 period. The findings are robust to a battery of unit root and cointegration tests. They support the Newhouse [Journal of Economic Perspectives 6 (1992) 3] conjecture that technological change is a major escalator of health care expenditure and confirm a significant and stable long-run relationship among per capita real health care expenditure, per capita real income and broad-based R&D expenditures. Policy implications are noted. PMID- 11852913 TI - An analysis of human response to the irritancy of acetone vapors. AB - Studies on the irritative effects of acetone vapor in humans and experimental animals have revealed large differences in the lowest acetone concentration found to be irritative to the respiratory tract and eyes. This has brought on much confusion in the process of setting occupational exposure limits for acetone. A literature survey was carried out focusing on the differences in results between studies using subjective (neuro)behavioral methods (questionnaires) and studies using objective measurements to detect odor and irritation thresholds. A critical review of published studies revealed that the odor detection threshold of acetone ranges from about 20 to about 400 ppm. Loss of sensitivity due to adaptation and/or habituation to acetone odor may occur, as was shown in studies comparing workers previously exposed to acetone with previously unexposed subjects. It further appeared that the sensory irritation threshold of acetone lies between 10,000 and 40,000 ppm. Thus, the threshold for sensory irritation is much higher than the odor detection limit, a conclusion that is supported by observations in anosmics, showing a ten times higher irritation threshold level than the odor threshold found in normosmics. The two-times higher sensory irritation threshold observed in acetone-exposed workers compared with previously nonexposed controls can apart from adaptation be ascribed to habituation. An evaluation of studies on subjectively reported irritation at acetone concentrations < 1000 ppm shows that perception of odor intensity, information bias, and exposure history (i.e., habituation) are confounding factors in the reporting of irritation thresholds and health symptoms. In conclusion, subjective measures alone are inappropriate for establishing sensory irritation effects and sensory irritation threshold levels of odorants such as acetone. Clearly, the sensory irritation threshold of acetone should be based on objective measurements. PMID- 11852915 TI - Cytosolic phospholipase A2 and lipoxygenase are involved in cell cycle progression in neuroblastoma cells. AB - Arachidonic acid has been implicated in regulating cellular proliferation, and is preferentially released by the 85-kDa cytosolic phospholipase A2 (cPLA2). Recently, we demonstrated that cPLA2 is activated at distinct periods during the ongoing cell cycle of neuroblastoma cells. The purpose of the present study was to establish the role of these cPLA2 activity peaks in cell cycle progression. Inhibition of cPLA2 activity with arachidonyl trifluoromethylketone (ATK) in early G1 phase reduced DNA synthesis markedly. A 24-h incubation with ATK revealed no significant difference in cell number compared to untreated cells, although cPLA2 activity was still inhibited. This suggests redundancy of different PLA2 enzymes. Lipoxygenase inhibition in early G1 resulted in G1 phase arrest, whereas inhibitors for cyclooxygenase had no effect. Furthermore, cells stopped progressing through S phase when lipoxygenase was inhibited in early S phase, demonstrating the requirement of lipoxygenase products for S phase progression. PMID- 11852914 TI - The mitochondrial PHB complex: roles in mitochondrial respiratory complex assembly, ageing and degenerative disease. AB - Although originally identified as putative negative regulators of the cell cycle, recent studies have demonstrated that the PHB proteins act as a chaperone in the assembly of subunits of mitochondrial respiratory chain complexes. The two PHB proteins, Phblp and Phb2p, are located in the mitochondrial inner membrane where they form a large complex that represents a novel type of membrane-bound chaperone. On the basis of its native molecular weight, the PHB-complex should contain 12-14 copies of both Phblp and Phb2p. The PHB complex binds directly to newly synthesised mitochondrial translation products and stabilises them against degradation by membrane-bound metalloproteases belonging to the family of mitochondrial triple-A proteins. Sequence homology assigns Phb1p and Phb2p to a family of proteins which also contains stomatins, HflKC, flotillins and plant defence proteins. However, to date only the bacterial HflKC proteins have been shown to possess a direct functional homology with the PHB complex. Previously assigned actions of the PHB proteins, including roles in tumour suppression, cell cycle regulation, immunoglobulin M receptor binding and apoptosis seem unlikely in view of any hard evidence in their support. Nevertheless, because the proteins are probably indirectly involved in ageing and cancer, we assess their possible role in these processes. Finally, we suggest that the original name for these proteins, the prohibitins, should be amended to reflect their roles as proteins that hold badly formed subunits, thereby keeping the nomenclature already in use but altering its meaning to reflect their true function more accurately. PMID- 11852916 TI - Progressive nodular histiocytosis in a child with a hypothalamic tumor. AB - We report a 13-year-old girl with multiple cutaneous histiocytic lesions, precocious puberty, growth hormone deficiency and a hypothalamic tumour. We conclude that she has progressive nodular histiocytosis, but this case illustrates the difficulty in differentiating the type II histiocytoses. PMID- 11852917 TI - The new EURO releases nickel and elicits contact eczema. PMID- 11852918 TI - Hobnail haemangioma occurring on the nasal dorsum. PMID- 11852919 TI - Isolated plantar collagenoma. PMID- 11852920 TI - Increased expression of alpha-melanocyte-stimulating hormone in the lesional skin of melasma. PMID- 11852921 TI - Kikuchi's disease, skin and systemic lupus erythematosus. PMID- 11852922 TI - Oral pathergy test in Behcet's disease. PMID- 11852923 TI - Sarcoidosis presenting with pyoderma granulosum. PMID- 11852924 TI - Immunohistochemical analysis of transforming growth factor beta 3 expression in solitary morphoea profunda with histological membranocystic changes. PMID- 11852925 TI - Abstracts from the Society for Cardiovascular Magnetic Resonance 5th annual scientific sessions. Lake Buena Vista, Florida, USA. January 25-27, 2002. PMID- 11852926 TI - Benign intracranial hypertension associated with arteriovenous malformation. AB - This is a report of a 9-year-old female with a clinical presentation of benign intracranial hypertension (BIH) who was found to have an unruptured pial arteriovenous malformation (AVM) with a significant fistula. The AVM was completely embolized using a recently developed liquid embolic system, Onyx, after which gradual clinical improvement followed. A few cases of BIH associated with AVM have been described in adults and adolescents. Possible causal relation is discussed. PMID- 11852927 TI - Systemic hypotension and white-matter damage in preterm infants. AB - This study was designed to test the hypothesis that systemic hypotension during the first postnatal week increases the risk of ultrasonographic echolucency in the white matter of preterm infants (< or = 28 weeks' gestation) while adjusting for confounders. From a study base of 1607 very-low-birthweight neonates (500 to 1500 g), a subsample of 243 preterm infants (122 females; < or = 28 weeks' gestation) was selected for echolucency and data collection prospectively for the entire first postnatal week. Data analyses were performed separately for the first 24 hours of life, for the interval from the end of the first 24 hours to the end of the fourth postnatal day, and for days 5, 6, and 7. Systemic hypotension was defined as the mean arterial blood pressure in the lowest quartile for the infant's week of gestational age. Protocol cranial ultrasounds were those obtained closest to days 1, 7, and 21. A committee of sonologists classified the infants as having either echolucency (echolucency group) or not (control group). Systemic hypotension during the first week of life appeared to be associated with echolucency in univariable analyses but the association did not persist after adjustment for potential confounders. Detailed summaries of 13 previous studies, the majority of which did not show an association between systemic hypotension and white-matter damage, are presented. In sum, these results do not support the hypothesis that systemic hypotension contributes to echolucency among preterm infants. PMID- 11852928 TI - Modified Sugiura procedure. PMID- 11852929 TI - Marine reserves and fisheries management. PMID- 11852930 TI - Marine reserves and fisheries management. PMID- 11852931 TI - Marine reserves and fisheries management. PMID- 11852932 TI - Long road ahead for hydrogen fuel cell cars. PMID- 11852933 TI - What counts in conservation? PMID- 11852934 TI - Mechanisms of traumatic brain injury: biomechanical, structural and cellular considerations. AB - Traumatic brain injury (TBI) is a public health problem of great concern, because it affects more than 2 million individuals each year. TBI occurs as a result of motor vehicle crashes, falls, and sports-related events. Biomechanical mechanisms occurring at the time of the injury initiate primary and secondary injuries that evolve over several days. In this article the relationship between an blunt injury event and the subsequent damage produced is addressed. Mechanisms of brain injury from biomechanics to cellular pathobiology are presented. Primary and secondary injuries are differentiated, and specific focal and diffuse clinical syndromes are described. Cellular mechanisms responsible for injury are also addressed, because they provide the unifying concepts across the many clinical syndromes so often discussed separately in reviews of traumatic brain injury. PMID- 11852936 TI - Application of technology in the treatment of traumatic brain injury. AB - The goal of care of the traumatic brain-injured patient is to prevent secondary injury. Technology gives the caregivers information as to the cause and severity of injury and can guide appropriate management of the patient. Use of multimodality monitoring increases the complexity of care but may allow for better targeted therapy. This article will discuss the current state of technology, the physiology and pathophysiology that it assesses, the normal and abnormal values obtained, and how care will be impacted. PMID- 11852935 TI - Pathophysiology of traumatic brain injury. AB - Approximately 7 million people in the United States each year experience traumatic brain injury, the leading cause of death from injury in this country. The initial traumatic event, often described as the primary injury, and the pathophysiologic sequelae, the secondary injury, are the themes of this article. Primary injuries to the extracranial and intracranial structures will be discussed, as well as the secondary injuries that occur minutes to days after the initial insult. It is critical that the bedside nurse clinician understand these concepts to anticipate potential complications and plan appropriate interventions with the multidisciplinary team. The goal of these interventions is to prevent the devastating effects of secondary injury. PMID- 11852937 TI - Care of the pediatric patient with brain injury in an adult intensive care unit. AB - Numerous differences exist between the pediatric and adult brain-injured patient. However, children with brain injuries are frequently cared for in adult Intensive Care Units (ICUs). This article is primarily designed for the nurse caring for the pediatric brain-injured patient in an adult ICU; however, because it is a current review, it can be useful to the pediatric practitioner. The article examines the care of the brain-injured child, including monitoring, psychosocial considerations, and supportive care with emphasis on avoiding secondary brain injury by decreasing and maintaining intracranial pressure. Differences between the pediatric patients and the adult patient are highlighted throughout. PMID- 11852938 TI - Head trauma in the older adult population. AB - The elderly person who is brain injured requires some special considerations because of the effects of aging on the body and brain. These individuals are increasingly admitted to the intensive care units for trauma from accidents, falls, and so forth. In addition to the care for the problems associated with brain trauma, the nurse must also be aware of the patient's medical history to prevent complications related to drug interactions, fluid overload, or volume deficit. This article addresses these considerations. PMID- 11852939 TI - Neuropsychological assessment in traumatic brain injury. AB - The care of patients provided in trauma centers involves many different variables. One of the complicating factors is related to the cognitive and emotional-behavioral consequences of traumatic brain injury. Impairment in attention, memory, communication skills, or planning and problems solving may present significant impediments in the ability of the patient to participate in his or her health care. In addition, the presence of emotional consequences such as anxiety, depression, irritability, and impulsivity may interfere with the ability of the patient to interact successfully with health care personnel. The role of the neuropsychologist is to provide a clinical assessment of the relevant cognitive and emotional problems areas and make suggestions for ways of interacting with and intervening with the brain-injured patient. PMID- 11852940 TI - Prescription discounts for health centers. PMID- 11852941 TI - Privacy task forces and commissions in the states. PMID- 11852942 TI - Predictions of secondary neutrons and their importance to radiation effects inside the International Space Station. AB - As part of a study funded by NASA MSFC to assess thecontribution of secondary particles in producing radiation damage to optoelectronics devices located on the International Space Station (IS), Monte Carlo calculations have been made to predict secondary spectra vs. shielding inside ISS modules and in electronics boxes attached on the truss (Armstrong and Colborn, 1998). The calculations take into account secondary neutron, proton, and charged pion production from the ambient galactic cosmic-ray (GCR) proton, trapped proton, and neutron albedo environments. Comparisons of the predicted neutron spectra with measurments made on the Mir space station and other spacecraft have also been made (Armstrong and Colborn, 1998). In this paper, some initial results from folding the predicted neutron spectrum inside ISS modules from Armstrong and Colborn (1998) with several types of radiation effects response functions related to electronics damage and astronaut-dose are given. These results provide an estimate of the practical importance of neutrons compared to protons in assessing radiation effects for the ISS. Also, the important neutron energy ranges for producing these effects have been estimated, which provides guidance for onboard neutron measurement requirements. PMID- 11852943 TI - Neutron measurements onboard the space shuttle. AB - The radiation environment inside a shielded volume is highly complex, consisting of both charged and neutral particles. Since the inception of human space flights, the charged particle component has received virtually all of the attention. There is however, a significant production of secondary neutrons, particularly from the aluminum structure in low earth orbiting spacecrafts. The interactions of galactic cosmic rays (GCR), and solar energetic particles with the earth's atmosphere produce a non-isotropic distribution of albedo neutrons. Inside any reasonable habitable module, the average radiation quality factor of neutrons is about 4-5 times larger than the corresponding average quality factor of charged particles. The measurement of neutrons and their energy spectra is a difficult problem due the intense sources of charged particles. This paper reviews the results of Shuttle flight experiments (made during both solar maximum and solar minimum) to measure the contribution of neutrons to the dose equivalent, as well as theoretical calculations to estimate the appropriate range of neutron energies that contribute most to the dose equivalent. PMID- 11852944 TI - Measurements of the high energy neutron component of cosmic radiation fields in aircraft using etched track dosemeters. AB - Measurements of the complex cosmic radiation field in aircraft at altitude are made with a passive survey meter comprising routine-use thermoluminescent detectors and etched track detectors. The energy dependence of response of the etched track detectors used to determine the neutron component has been characterized, partly, up to a neutron energy of 180 MeV. The neutron detectors are routinely calibrated in the CERN/EC Reference Field. The 15% determination level for total dose equivalent is 100 microSv. The evidence is that the passive survey meter provides a reliable determination of route dose. PMID- 11852946 TI - Kass v. Kass. PMID- 11852945 TI - Neutron dosimetry in low-earth orbit using passive detectors. AB - This paper summarizes neutron dosimetry measurements made by the USF Physics Research Laboratory aboard US and Russian LEO spacecraft over the past 20 years using two types of passive detector. Thermal/resonance neutron detectors exploiting the 6Li(n,T) alpha reaction were used to measure neutrons of energies <1 MeV. Fission foil neutron detectors were used to measure neutrons of energies above 1 MeV. While originally analysed in terms of dose equivalent using the NCRP 38 definition of quality factor, for the purposes of this paper the measured neutron data have been reanalyzed and are presented in terms of ambient dose equivalent. Dose equivalent rate for neutrons <1 MeV ranged from 0.80 microSv/d on the low altitude, low inclination STS-41B mission to 22.0 microSv/d measured in the Shuttle's cargo bay on the highly inclined STS-51F Spacelab-2 mission. In one particular instance a detector embedded within a large hydrogenous mass on STS-61 (in the ECT experiment) measured 34.6 microSv/d. Dose equivalent rate measurements of neutrons >1 MeV ranged from 4.5 microSv/d on the low altitude STS 3 mission to 172 microSv/d on the ~6 year LDEF mission. Thermal neutrons (<0.3 eV) were observed to make a negligible contribution to neutron dose equivalent in all cases. The major fraction of neutron dose equivalent was found to be from neutrons >1 MeV and, on LDEF, neutrons >1 MeV are responsible for over 98% of the total neutron dose equivalent. Estimates of the neutron contribution to the total dose equivalent are somewhat lower than model estimates, ranging from 5.7% at a location under low shielding on LDEF to 18.4% on the highly inclined (82.3 degrees) Biocosmos-2044 mission. PMID- 11852947 TI - Acute care management of severe traumatic brain injuries. AB - Preservation or restoration of optimal neurologic function following traumatic brain injury (TBI) requires timely and aggressive therapeutic interventions. Effective diagnostic tools, together with an armamentarium of treatment modalities, have augmented the treatment strategies utilized today. In addition, the Guidelinesfor the Management of Severe Head Injury have established a standardized approach for the TBI patient. This article will provide current information regarding the resuscitation priorities, appropriate interventions, and pharmacological agents used in the treatment required by the complex nature of TBI. Also, a review of the occurrences associated with TBI will be discussed. PMID- 11852948 TI - Traumatic brain injury research: a review of clinical studies. AB - There is a growing volume of research on trauma brain injury (TBI) as evidenced by a recent Medline search that reported over 6000 articles published on TBI in the past 5 years. PMID- 11852949 TI - Measuring quality of life of patients of traumatic brain injury. AB - Quality of life (QOL) is recognized as an important indicator of health care and the patient's ability to cope with illness, treatment, and recuperation. Issues that need to be addressed in any proposed QOL research include a clear definition of QOL, a sound rationale for the choice of a measurement instrument, and the value of qualitative data. Measuring QOL in a patient population that has experienced traumatic brain injury (TBI) raises special concerns associated with the physical, behavioral, and cognitive limitations inherent with the specific TBI population. These pertinent issues are discussed with a focus that should be helpful for persons planning QOL projects and those reading and critiquing related literature. A study conducted by the author with patients with severe trauma injury will be used as an example of the impact of these issues on an actual project. PMID- 11852950 TI - The history of the Glasgow Coma Scale: implications for practice. AB - The Glasgow Coma Scale (GCS) has been the gold standard of neurologic assessment for trauma patients since its development by Jennett and Teasdale in the early 1970s. The GCS was found to be a simple tool to use. It became the method of choice for trauma care practitioners to document neurologic findings over time and predict functional outcome. Although the scale has been shown to be effective, many authors have cited weaknesses in the scale including the inability to predict outcome, variation in inter-rater reliability, and the inconsistent use by caregivers in the prehospital and hospital settings. This article outlines the components of the GCS and how practitioners can best use the scale, particularly in patients whose injuries and treatments make them difficult to assess. PMID- 11852951 TI - Critical incidents: effective communication and documentation. AB - Critical incidents can be defined at sentinel events, critical patient care issues, or any patient event outside the normal parameters of care. Nurses are central to maintaining the standard of care for patients in these situations. Miscommunication, including inadequate communication and illegible or incomplete documentation, forms a basis for many clinical and interpersonal problems in nursing practice. This practical article discusses expectations, both professional and legal, for avoiding and/or managing critical incidents. Suggestions for assessment parameters to report are included in table format, which the nurse can 'clip and use' as a reminder of data to have prepared before calling and/or presenting the problem. Steps are proposed for a logical, organized plan to help the nurse communicate more effectively. Recommendations for essentials of documentation regarding the verbal interaction and orders received are also presented. PMID- 11852952 TI - Present status of cardiac pacing: a nursing perspective. AB - In this article, we focus on the nurse's role as caregiver and patient advocate for individuals requiring cardiac pacing. The history of cardiac pacing, new trends, indications for pacing, pacemaker dysfunction and complications, types of pacing, and nursing considerations will be discussed. PMID- 11852953 TI - Acute dissection of the aorta: a review for nurses. AB - This article will review important elements involved in caring for the patient experiencing an acute dissection of the aorta. The etiology and classification system, signs and symptoms, diagnostic methods, indications for operation, circulatory arrest/deep hypothermia, surgical techniques, and complications of surgery will be discussed. PMID- 11852954 TI - Radial artery: an alternative revascularization conduit. AB - In the early 1970s, patients underwent myocardial revascularization with a radial artery graft. Soon after its introduction as an additional arterial conduit, postprocedural complications were observed, and the use of this conduit was abandoned. A resurgence of the radial artery as a free graft was seen 20 years later. This article will discuss the background, renewing interests in the use of the radial artery, selection of candidates, perioperative preparation, medical/nursing considerations, and postoperative outcomes from an interdisciplinary approach. PMID- 11852955 TI - Endovascular aortic aneurysm repair: an alternative approach. AB - Endovascular aneurysm repair is evolving technology that has presented the patient and physician with an alternative surgical approach. Preoperative planning, graft selection, and comprehensive imaging are critical factors required for successful endovascular abdominal aortic repair. This article will address preoperative planning, endovascular techniques, complications, postoperative management, and benefits from the procedure. PMID- 11852956 TI - Minimally invasive saphenous vein harvesting. AB - The intent of this article is to describe the emergence and continuing evolvement and improvement in harvesting saphenous veins by the minimally invasive versus the traditional open technique. The comparison of endoscopic versus open techniques, patient population, systems limitations, clinical outcomes, and perioperative care of the patient will be discussed. The author's intent is to take a multidisciplinary approach. PMID- 11852957 TI - Case study: a patient's survival. AB - Presentation of a case study involving a female patient, in her 20s, undergoing routine surgery for removal of atrial myxoma leading to a heart transplant. This case study will show the progression from postcardiotomy failure, the emergent use of the extracorporeal membrane oxygenator device, the insertion of the HeartMate device, and the final return to the operating room for a heart transplant. The case study will examine the physiologic demands on the patient, as well as the psychological effects from the various life-saving devices. PMID- 11852958 TI - Minimally invasive coronary artery bypass surgery. AB - Advances in videoscopic instrumentation and expanding experience with the performance of coronary artery bypass surgery without cardiopulmonary circulatory support is changing the surgical approach to many patients requiring coronary arterial revascularization. We describe the present status of minimally invasive coronary artery bypass surgery being used today. PMID- 11852959 TI - Minimally invasive aortic valve surgery. AB - Heart surgery has seen the emergence of minimally invasive surgery in the quest for less traumatic and less painful surgery. This procedure can be provided without the increased cost of endoscopic instrumentation by use of standard instrumentation, cannulation, and prostheses. A small incision and partially split sternum provide the patient with less postoperative pain and bleeding and early extubation and discharge from the hospital. PMID- 11852960 TI - The effects of hypothermia on coronary artery bypass graft surgery. AB - This article will discuss how induced hypothermia affects the patient undergoing coronary artery bypass graft surgery. Nursing interventions differ greatly for cardiac procedures done on cardiopulmonary bypass versus off-bypass procedures, such as "keyhole." Induced hypothermia, cardiopulmonary bypass, cardioplegia, and various delivery techniques of cardioplegia, and postoperative hypothermia will be discussed. PMID- 11852961 TI - Metabolic basis of drug interactions in the intensive care unit. AB - Drug interactions are a potentially hazardous consequence of drug therapy, particularly in the critically ill. Many drug interactions result from changes in drug metabolism. These interactions can result in either an increase in toxicity or a decrease in effect. The process of normal drug metabolism depends on the activity of individual metabolizing enzymes. The activity of these enzymes can be altered by many factors, and it is alterations in these enzymes that account for most drug interactions. This article reviews normal and abnormal drug metabolism and discusses how this knowledge can be used to predict and prevent potential drug interactions. PMID- 11852962 TI - Monitoring drug therapy in the intensive care unit. AB - Drugs are among the most commonly administered therapeutic interventions received. In the intensive care unit (ICU), patients routinely receive more therapies than on general medical or surgical wards, and practitioners caring for these patients are presented with the challenge of monitoring each of these therapies for efficacy and toxicity. The goal of this article is to describe a conceptual approach that practitioners can use to monitor for drug efficacy and predictable and unpredictable adverse drug effects. Although therapeutic drug monitoring (TDM) assumes a relatively small role in the overall monitoring scheme, TDM is discussed because it serves an important function for a select group of agents. PMID- 11852963 TI - Review of common adverse effects of selected antiarrhythmic drugs. AB - The management of cardiac arrhythmias has changed dramatically over the past several years. New drugs and devices are now available to treat various arrhythmias. Many new agents have been developed that rely on different electrophysiologic mechanisms to elicit their effect on the heart rhythm. Though often effective, these drugs also pose a risk because all of them have a variety of potential adverse effects associated with their use. Many of these adverse reactions are common to all antiarrhythmic drugs, whereas others are unique to particular agents. This review discusses the notable adverse effects of selected antiarrhythmic drugs. PMID- 11852964 TI - Stress ulcer prophylaxis in intensive care patients: an evidence-practice mismatch? AB - Prophylactic drugs are commonly used in intensive care units (ICUs) to decrease the risk of development of stress-related upper gastrointestinal tract lesions and bleeding. The aim of this article is to discuss history, incidence, risk factors, past and current research, and current evidence for usefulness of prophylaxis for this disorder. Currently, very good evidence suggests that most ICU patients do not benefit from the widespread application of prophylactic intervention. PMID- 11852965 TI - Delirium in the intensive care unit: are we helping the patient? AB - The intensive care unit (ICU) represents a dynamic interaction between patient factors and interventional factors. The complexity of this situation can generate an impaired consciousness in the patients. The critical care provider is faced with deducing the etiology and treatment of delirium in the ICU. Many of the therapeutic agents that are used in the ICU may precipitate delirium. Patients may also experience delirium as part of their underlying medical conditions. Withdrawal syndromes, delirium tremens in particular, are known to cause delirium. By a combination of appropriate selection of medications and an awareness of delirium as a side effect, the patient in the ICU may be treated in a manner to minimize the clouding of consciousness. An understanding of the proposed pathophysiology of various types of delirium will allow appropriate clinical measures to be taken. PMID- 11852966 TI - New antithrombotics for the intensive care unit setting: GP IIb/IIIa inhibitors, low-molecular-weight heparins, and direct thrombin inhibitors. AB - Thromboembolic disease (TED) is a frequent problem encountered by clinicians in the intensive care unit (ICU). Traditionally, unfractionated heparin (UFH) has been used in the treatment and prophylaxis of TED. However, newer antithrombotic agents have evolved as effective alternatives to UFH. Low-molecular-weight heparins are effective for both the treatment and prophylaxis of TED. Lepirudin is useful for patients with TED and a history of heparin-induced thrombocytopenia. The platelet glycoprotein IIb/IIIa inhibitors are synergistic with UFH for the treatment of acute coronary syndromes and percutaneous coronary interventions (PCI). Several drugs are now available to clinicians for the treatment and prophylaxis of TED. PMID- 11852967 TI - Nutrition support in the critically ill patient. AB - Despite an absence of well-controlled studies demonstrating a clear mortality benefit, providing nutrition support in the critically ill patient has become routine in most ICU settings. Unless clearly contraindicated, patients should be fed enterally, using conventional isotonic feedings employing gastric or postpyloric access. When to begin nutrition support varies, depending on baseline nutritional status, anticipated time until oral feedings are resumed, and the degree of stress. Energy and protein requirements should be assessed routinely with minimum goals of avoiding overfeeding and minimizing any net negative nitrogen balance. All patients receiving feedings require close surveillance to identify predictable complications and to tailor therapy to achieve nutritional goals. Adjunctive therapies should be employed as needed to help achieve nutritional goals, eg, insulin infusions to control serum glucose and prokinetic agents to improve gastric emptying. When feasible and safe, parenterally fed patients should be transitioned to enteral or oral feedings when appropriate. PMID- 11852968 TI - Sedation of the mechanically ventilated patient. AB - Goals in the care of the mechanically ventilated patient are sedation, analgesia, anxiolysis, and muscle relaxation. Causes of distress in these patients include: pain, sleep deprivation, anxiety, psychosis, agitation, and delirium. Drugs used to alleviate these stressors are opiates, benzodiazepines, neuromuscular blocking agents, anesthetic induction agents, and inhalational agents. When caring for the agitated patient on the mechanical ventilator, physiologic, mechanical, and emotional causes must all be investigated. Finally, nonpharmacologic therapy is of utmost importance in the care of these patients. PMID- 11852969 TI - The role of cytokines in regulating protein metabolism and muscle function. AB - Multiple lines of evidence suggest that cytokines influence different physiologic functions of skeletal muscle cells, including anabolic and catabolic processes and programmed cell death. Cytokines play an important role not only in muscle homeostasis, therefore, but also in the pathogenesis of different relevant clinical conditions characterized by alterations in protein metabolism. Recently discovered cytokines, such as ciliary neurotrophic factor and growth/differentiation factor-8, as well as the more studied tumor necrosis factor-alpha, interleukin-1, interleukin-6, and the interferons, have been implicated in the regulation of muscle protein turnover. Their postreceptor signaling pathways, proteolytic systems, and the mechanisms of protein synthesis inhibition involved in different catabolic conditions have been partially clarified. Moreover, recent studies have shown that cytokines can directly influence skeletal muscle contractility independent of changes in muscle protein content. Even though several gaps remain in our understanding, these observations may be useful in the development of strategies to control protein metabolism and muscle function in different clinical conditions. PMID- 11852970 TI - Fortification strategies to combat zinc and iron deficiency. AB - Food fortification is an important strategy to combat iron and zinc deficiency. This review covers the basic concepts of food fortification, as well as its advantages and disadvantages. The main characteristics of the most common zinc and iron compounds used in this procedure are also analyzed. PMID- 11852971 TI - Malnutrition and excess mortality in Shangri-La. AB - Populations living at high altitude in the Tibetan highlands suffer extraordinarily high rates of maternal mortality, infant and juvenile mortality, and infectious morbidity. Poverty and living condition, more than altitude, contribute to the adverse statistics. The traditional nomadic herders are the most affected among the residents. The question remains whether contemporary standards of adequate health and nutrition are compatible with the integral folkways of traditional tribal groups. PMID- 11852972 TI - A newly discovered visual cycle necessary for vision during continuous illumination. AB - Rhodopsin, the light-sensitive visual pigment of the retina, is activated through photoisomerization of its prosthetic group, 11-cis-retinal, to all-trans-retinal. A protein found in the retinal pigment epithelium named retinal G protein- coupled receptor (RGR) reacts to light in an opposite but parallel way: its prosthetic group, all-trans-retinal, is photoisomerized to 11-cis-retinal. The latter is reduced to 11-cis-retinol by a cis-retinol dehydrogenase that co purifies with the RGR. The resulting 11-cis-retinol feeds into the visual cycle to be oxidized to 11-cis-retinal, thus replenishing the 11-cis-retinal of the rhodopsin. During continuous intense illumination, RGR can supplement the 11-cis retinal required to regenerate rhodopsin. PMID- 11852973 TI - Cyclooxygenase inhibition as a strategy to ameliorate brain injury. AB - Cyclooxygenase (COX) is the obligate, rate-limiting enzyme for the conversion of arachidonic acid into prostaglandins. Two COX enzymes have been identified: a constitutively expressed COX-1 and an inducible, highly regulated COX-2. Widely used to treat chronic inflammatory disorders, COX inhibitors have shown promise in attenuating inflammation associated with brain injury. However, the use of COX inhibition in the treatment of brain injury has met with mixed success. This review summarizes our current understanding of COX expression in the central nervous system and the effects of COX inhibitors on brain injury. Three major targets for COX inhibition in the treatment brain injury have been identified. These are the cerebrovasculature, COX-2 expression by vulnerable neurons, and the neuroinflammatory response. Evidence suggests that given the right treatment paradigm, COX inhibition can influence each of these three targets. Drug interactions and general considerations for administrative paradigms are also discussed. Although therapies targeted to specific prostaglandin species, such as PGE2, might prove more ameliorative for brain injury, at the present time non specific COX inhibitors and COX-2 specific inhibitors are readily available to researchers and clinicians. We believe that COX inhibition will be a useful, ameliorative adjunct in the treatment of most forms of brain injury. PMID- 11852974 TI - Guidelines for prehospital management of traumatic brain injury. PMID- 11852975 TI - Misclassification and treatment effect on primary outcome measures in clinical trials of severe neurotrauma. AB - The power of clinical trials depends mainly on the choice of the primary outcome measure, the statistical test, and the sample size. The most widely used outcome measure has been the five-category Glasgow Outcome Scale (GOS). Contrary to intuition, we show that more categories do not necessarily increase the power of a trial and actually can decrease power. This is so for two reasons. The more categories of outcome measure used, the more the likelihood for misclassifications. The effect of 0%, 10%, and 20% misclassification rate upon power is illustrated. Misclassification rates in two completed trials are examined based on comparative overlap in GOS and Disability Rating Scale (DRS) categories. The outcome results of the "National Acute Brain Injury Study: Hypothermia" indicate that the ideal number of categories also depends upon the effect of study treatment. In the recently completed hypothermia trial, the use of a dichotomized GOS (good recovery/moderate disability versus severe disability/vegetative/dead) is shown to be more sensitive than use of three or more categories of the GOS. The results point to the importance of training study investigators who will collect the outcome data. The results also indicate that the number of categories should be carefully determined using the pilot data or the data from phase II trials. PMID- 11852977 TI - Differential effect of spinal cord injury and functional impairment on human brain activation. AB - Reorganization of human brain function after spinal cord injury (SCI) has been shown in electrophysiological studies. However, it is less clear how far changes of brain activation in SCI patients are influenced by the extent of SCI (neuronal lesion) or the consequent functional impairment. Positron emission tomography ([15O]-H2O-PET) was performed during an unilateral hand movement in SCI patients and healthy subjects. SCI patients with paraplegia and normal hand function were compared to tetraplegic patients with impaired hand movements. Intergroup comparison between paraplegic patients and healthy subjects showed an increased activation of contralateral sensorimotor cortex (SMC), contralateral thalamus, ipsilateral superior parietal lobe, and bilateral cerebellum. In contrast to this, tetraplegic patients with impaired upper limb function revealed only a significant activation of supplementary motor area (SMA). Correlational analysis in the tetraplegic patients showed that the strength of hand movement was related to the activation of contralateral SMC. However, the severity of upper limb sensorimotor deficit was related to a reduced activation of contralateral SMA and ipsilateral cerebellum. The findings suggest that in paraplegic patients with normal hand function the spinal neuronal lesion itself induces a reorganization of brain activation unrelated to upper limb function. Compared to this, in tetraplegic patients changes of brain activation are related to the impaired upper limb function. Therefore, in patients with SCI a differential impact of spinal lesion and functional impairment on brain activation can be shown. The effect of impaired afferent feedback and/or increased compensatory use of non impaired limbs in SCI patients needs further evaluation. PMID- 11852976 TI - Group I metabotropic glutamate receptors in spinal cord injury: roles in neuroprotection and the development of chronic central pain. AB - Spinal cord injury (SCI) initiates a cascade of biochemical events that leads to an increase in extracellular excitatory amino acid (EAA) concentrations, which results in glutamate receptor-mediated excitotoxic events. An important division of these glutamate receptors is the metabotropic glutamate receptor (mGluR) class, which is divided into three groups. Of these three groups, group I (mGluR1 and mGluR5) activation can initiate a number of intracellular pathways that lead to increased extracellular EAA concentrations. To evaluate subtypes of group I mGluRs in SCI, we administered AIDA (group I antagonist), LY 367385 (mGluR1 specific antagonist), or MPEP (mGluR5 specific antagonist) by interspinal injection to adult male Sprague-Dawley rats (175-200 g) immediately following injury at T10 with an NYU impactor (12.5-mm drop, 10-g rod, 2 mm in diameter). AIDA- and LY 367385-treated subjects had improved locomotor scores and demonstrated an attenuation in the development of mechanical allodynia as measured by von Frey stimulation of the forelimbs; however, LY 367385 potentiated the development of thermal hyperalgesia. MPEP had no effect on locomotor recovery or mechanical allodynia, but attenuated the development of thermal hyperalgesia. AIDA and LY 367385 treatment resulted in a significant increase in tissue sparing compared to the vehicle-treated group at 4 weeks following SCI. These results suggest that mGluRs play an important role in EAA toxicity and have different acute pathophysiological roles following spinal cord injury. PMID- 11852978 TI - Sensorimotor cortical activity in patients with complete spinal cord injury: a functional magnetic resonance imaging study. AB - Residual activation of the cortex was investigated in nine patients with complete spinal cord injury between T6 and L1 by functional magnetic resonance imaging (fMRI). Brain activations were recorded under four conditions: (1) a patient attempting to move his toes with flexion-extension, (2) a patient imagining the same movement, (3) passive proprio-somesthesic stimulation of the big toes without visual control, and (4) passive proprio-somesthesic stimulation of the big toes with visual control by the patient. Passive proprio-somesthesic stimulation of the toes generated activation posterior to the central sulcus in the three patients who also showed a somesthesic evoked potential response to somesthesic stimulation. When performed under visual control, activations were observed in two more patients. In all patients, activations were found in the cortical areas involved in motor control (i.e., primary sensorimotor cortex, premotor regions and supplementary motor area [SMA]) during attempts to move or mental imagery of these tasks. It is concluded that even several years after injury with some local cortical reorganization, activation of lower limb cortical networks can be generated either by the attempt to move, the mental evocation of the action, or the visual feedback of a passive proprio-somesthesic stimulation. PMID- 11852979 TI - Bcl-2 and GDNF delivered by HSV-mediated gene transfer after spinal root avulsion provide a synergistic effect. AB - Proximal spinal nerve injury results in the death of motor neurons in ventral horn. We have previously demonstrated this cell death can be prevented by HSV mediated transfer of the gene coding for the antiapoptotic peptide Bcl-2 7 days prior to injury, but that expression of Bcl-2 does not preserve ChAT expression in the lesioned cells. In the current study, we examined two related issues: whether Bcl-2 delivered by HSV-mediated gene transfer 30 min after injury could similarly protect motor neurons from cell death, and whether the additional HSV mediated expression of the glial cell derived neurotrophic factor (GDNF) could improve the result. At 30 min after avulsion of the L4, L5, and L6 spinal nerves, replication defective genomic HSV-based vectors coding for Bcl-2, GDNF, a reporter transgene (lacZ), or the Bcl-2 and GDNF vectors together were injected into spinal cord. Transduction of motor neurons with either the Bcl-2-expressing vector or the GDNF-expressing vector resulted in a substantial increase in the number of surviving motor neurons, and coinjection of the two vectors together resulted in cell survival that was similar to the result obtained with either vector alone. Neither the Bcl-2-expressing vector nor the GDNF-expressing vector delivered alone protected choline acetyltransferase (ChAT) expression in lesioned neurons. However, simultaneous injection of the Bcl-2- and the GDNF-expressing vectors together resulted in a substantial increase in the number of ChAT in cells in the lesioned ventral horn. Together, these findings suggest an approach to improving cell survival and regeneration following proximal root injury. PMID- 11852980 TI - Extradural compression of sensorimotor cortex: a useful model for studies on ischemic brain damage and neuroprotection. AB - Behavioral and morphological changes were examined for up to 9 days after moderate cerebral ischemia caused by slow compression of a specific brain area in the sensorimotor cortex of Sprague-Dawley rats. Functional deficits after the cerebral ischemia were assessed by daily beam-walking tests, whereas morphological changes were verified using Nissl staining on day 1, 2, 3, 5, and 9, respectively. Rats exposed to cerebral ischemia displayed impaired beam walking performance. Mild hypothermia prevented both the compression-produced functional deficits and the brain damage. Younger (5 weeks) animals showed less neurological deficits than older (9 weeks) animals. Histological examination revealed a pronounced increase in the number of injured pyramidal neurons from day 1 to day 3 in the primarily damaged brain region. Between day 3 and day 5, the number of injured cells remained constant, whereafter there was a slow decline of thionin-positive neurons as examined on day 9. The noncompetitive NMDA receptor antagonist, dizocilpine (MK-801; 3 mg/kg, i.p.), did not alter the neurological impairment on day 1, but improved thereafter the rate of functional recovery and reduced the number of damaged cells. The AMPA receptor antagonist, LY326325 (15 or 30 mg/kg; i.p.), dose-dependently diminished the neurological deficits on day 1, enhanced the rate of recovery, and reduced the number of injured neurons over time. Our data suggest that short-lasting extradural compression of a well-defined brain area in the sensorimotor cortex is a highly reproducible model with a high success rate for the study of functional and morphological consequences after cerebral ischemia as well as for the evaluation of the therapeutic potential of novel, neuroprotective pharmacological agents. PMID- 11852981 TI - Effects of global ischemia duration on neuronal, astroglial, oligodendroglial, and microglial reactions in the vulnerable hippocampal CA1 subregion in rats. AB - The hippocampal CA1 neurons are selectively vulnerable to global ischemia, and neuronal death occurs in a delayed manner. The threshold of global ischemia duration that induces neuronal death has been studied, but the relationship between ischemia duration and glial death in the hippocampal CA1 area has not been fully studied. We examined neuronal/glial viability and morphological changes in the CA1 subregion after different durations of global ischemia. Global ischemia was induced in Sprague-Dawley rats by 10, 5, and 3 min of bilateral common carotid artery occlusion and hypotension. At 1-56 days after ischemia, the morphological reactions of neurons, astrocytes, oligodendrocytes, and microglia were immunohistochemically evaluated. Most of the hippocampal CA1 pyramidal neurons underwent delayed death at 3 days after 10/5 min of ischemia, but not after 3 min of ischemia. The number of astrocytes gradually declined after 10/5 min of ischemia, and viable astrocytes showed characteristic staged morphological reactions. Oligodendrocytes also showed morphological changes in their processes after 10/5 min of ischemia. Microglia transformed into a reactive form at 5 days only after 10/5 min of ischemia. These data suggest that some morphological changes in glial cells were not dependent on neuronal cell death, but their own reactions to the different severity of ischemia. PMID- 11852982 TI - Axonal transection in adult rat brain induces transsynaptic apoptosis and persistent atrophy of target neurons. AB - We used the fimbria-fornix (FF) transection model of axonal injury to test the hypothesis that transneuronal degeneration occurs in the adult central nervous system in response to deafferentation. The medial mammillary nucleus, pars medialis (MMNm) was analyzed by light and electron microscopy at 3, 7, 14, and 30 days, and 6 months after unilateral FF transection in adult rat to identify the time course of neuronal responses in a remote target. Presynaptic terminals and neuronal cell bodies degenerated in the MMNm ipsilateral to FF transection. Terminal degeneration occurred predominantly at 3 and 7 days postlesion. Between 14 and 30 days postlesion, neuronal number in the MMNm decreased (approximately 20%). Two forms of neuronal degeneration were found in the MMNm after deafferentation. Some neurons died apoptotically. Other neurons underwent vacuolar degeneration. In these latter neurons, somatodendritic pathology occurred at 14 and 30 days and 6 months postlesion. The ultrastructure of this vacuolar degeneration was characterized by disorganization of the cytoplasm, formation of membrane-bound vacuolar cisternae and membranous inclusions, loss of organelles, cytoplasmic pallor, and chromatin alterations. This study shows that both anterograde axonal degeneration and transneuronal degeneration occur in a fornical target after FF axon transection. This transneuronal degeneration can be classified as sustained neuronal atrophy or transsynaptic apoptosis. PMID- 11852983 TI - Clinical education, circa 2010. PMID- 11852984 TI - Designing clinical examples to promote pattern recognition: nursing education based research and practical applications. AB - Despite carefully planned classroom sessions and richly designed clinical experiences, nurse educators know that, realistically, students will not encounter all of the life-threatening patient situations that require recognition and intervention during these sessions and experiences. However, on graduation, these students will be legally accountable for recognition of these situations and the immediate interventions associated with them. Therefore, the challenge for nurse educators is how to structure didactic interactions that promote pattern recognition in anticipation of such yet-to-be-experienced events. PMID- 11852985 TI - Baccalaureate nursing students' communication process in the clinical setting. AB - The purpose of this study was to describe and generate theory about how baccalaureate nursing students communicate with patients in the clinical setting using the qualitative method of grounded theory. Third-year and fourth-year generic baccalaureate nursing students were the primary participants in this study. Data were gathered during two academic semesters from formal and informal interviews and observations in hospital and home care settings. Transcribed audiotapes and field memos constituted the data generated in the study and were subjected to generating categories through open coding of data, applying theoretical codes to the data, collapsing and expanding categories reflective of the data, and memoing and uncovering the core category of the data. The core category then became the grounded theory. Analysis of the data revealed that students encountered a basic social psychological problem of "saying the right things to patients" in the clinical setting, which was preceded by the antecedent problem of "fear of saying the wrong things to patients." Students solved the problem, in part, by employing communication strategies and patterns. Eight communication strategies and five communication patterns incorporating communication strategies were identified. In addition, students went through a four-stage basic social psychological process. The process of learning a personal communication repertoire was uncovered as the substantive grounded theory. The repertoire was learned by students in the dynamic environment of the clinical setting in which RNs served as role models and communicating with difficult patients was the crucial step in the process of communication development. PMID- 11852986 TI - Development of the Clinical Learning Environment Inventory: using the theoretical framework of learning environment studies to assess nursing students' perceptions of the hospital as a learning environment. AB - Clinical learning is an integral part of nursing education; yet, clinical education has been problematic. Why are some clinical experiences better than others? Various studies have indicated that not all practice settings are able to provide student nurses with a positive learning environment. The clinical learning environment is a multidimensional entity with a complex social context. Classroom learning environment research based on psychosocial educational conceptual frameworks has been well established in the past 30 years, with evidential benefits. Previous research on clinical learning environment was examined. However, minimal studies have been conducted on hospital learning environments from the psychosocial educational perspective. To maximize nursing students' clinical learning experiences, the author developed the Clinical Learning Environment Inventory based on the theoretical framework in psychosocial education. PMID- 11852987 TI - Community collaboration: using nursing students in a smoking cessation program for pregnant women. PMID- 11852989 TI - Camp communities: valuable clinical options for BSN students. PMID- 11852990 TI - Nurturing staff-student relationships. PMID- 11852988 TI - Consistent clinical assignment for nursing students compared to multiple placements. PMID- 11852991 TI - Teaching strategy to maximize clinical experience with beginning nursing students. PMID- 11852992 TI - Mismatch repair and the hereditary non-polyposis colorectal cancer syndrome (HNPCC). AB - The hereditary non-polyposis colorectal cancer (HNPCC)-syndrome is the most common form of hereditary colorectal cancers, and accounts for 2-7% of the total colorectal cancer burden. Since there are no single clinical features specific for HNPCC, diagnosis is based on family history (Amsterdam or Bethesda criteria) and is confirmed by the detection of a mutation in one of the responsible mismatch repair (MMR) genes. Two types of HNPCC-families can be distinguished. Type I HNPCC tumors are exclusively located in the colon, whereas in Type II HNPCC patients, extracolonic tumors are present in the stomach, endometrium, ovary, and urinary tract. The identification of the human homologues of yeast mismatch repair genes hMSH2, hMSH3, hMSH6, hMLH1, hMLH3, hPMS1 (scMLH2), and hPMS2 (scPMS1) offered the prospect of genetic screening leading to an extensive search for mutations in HNPCC-families. The majority of the alterations have been found in hMSH2 (40%) and hMLH1 (40%) genes. Mutations in the other MMR genes appear rare, absent, and/or associated with atypical families (1-5%). As a result of the mismatch repair deficiency, replication misincorporation errors accumulate, resulting in a mutator phenotype. Diagnosis of HNPCC-associated replication errors is most easily determined by the examination of a panel of the National Cancer Institute (NCI)-recommended simple repeated sequences (microsatellites), combined with immunohistochemical analysis. Although the exact molecular mechanism of the tumor development in these patients remains poorly understood, the identification of tumors that harbor a microsatellite instability has clinical and prognostic implications. PMID- 11852994 TI - Marrow stem cells, mesenchymal progenitor cells, and stromal progeny. PMID- 11852993 TI - Heterogeneity of ovarian cancer: relationships among histological group, stage of disease, tumor markers, patient characteristics, and survival. AB - Epidemiological studies have established associations between various reproductive factors and risk of ovarian cancer; it has also been observed that some of these risk factors are only associated with specific histological subgroups. To investigate the correlation of genetic alterations with these risk factors, we examined a consecutive series of 158 ovarian cancer cases treated at the University of Kentucky (1990-96). Common molecular genetic alterations (LOH on chromosome 17, P53 alterations, K-RAS mutations), histological and clinical characteristics of the disease, demographic patient information and survival were evaluated. These latter data were from the Kentucky Cancer Registry. Univariate analysis showed higher frequencies of chromosome 17 loss and P53 mutations in tumors of advanced stage and grade, and in older and post-menopausal women. Non mucinous tumors were more likely to be classified as late stage, high-grade cancers, and to have chromosome 17 loss and P53 mutations. Survival analysis indicated that stage was the only independent significant variable. When stage was the outcome variable in multiple logistic regression analysis, histology and chromosome 17 loss were significantly associated with poor survival. This case case study provides evidence that ovarian cancers of mucinous and non-mucinous histology are significantly different with respect to clinical characteristics, survival and molecular alterations. It also lends support to the hypothesis that ovarian cancer is a heterogeneous disease with distinct etiological factors and clinical outcomes, which may require different approaches to treatment. PMID- 11852995 TI - Hematopoietic stem cells. PMID- 11852997 TI - The HTLV cell surface receptor. PMID- 11852998 TI - Experience with methotrexate, 5-fluorouracil, and leucovorin (MFL): a first line effective, minimally toxic regimen for metastatic breast cancer. AB - Thirty-two women with untreated metastatic breast cancer were treated with 100 mg/M2 i.v. methotrexate (MTX), 600 mg/M2 5-fluorouracil (5FU) and leucovorin 15 mg orally every 6 hr, 24 hr after MTX (MFL) on days 1 and 8 every 28 days. Stratification was according to sites of metastases (mets), adjuvant (adj), chemotherapy (CTX), and/or hormonal therapy or no adj therapy (Tx). Treatment continued until documented radiographic or clinical disease was in progression. Toxicity was mild, consisting of only minimal elevations of transaminases and mild cytopenias. There was no pulmonary toxicity. There were no hospitalizations, treatment delays or cessations for toxicity. One patient with skeletal mets had a complete response and 7 had partial responses. The overall median progression free survival (PFS) was 13.8 months (mos). Eighteen patients with skeletal mets had PFS from 7-70 mos (median 15.9). Five patients with lung mets only had PFS from 6-20 mos (median 9.8 mos). Patients with liver alone or with other visceral mets showed progression within 2-5 mos. However, patients with bone and visceral mets without liver involvement had PFS from 8-50 mos (median 20.5). Of 21 adj Tx failures the median PFS was 8.8 mos (2-94). Six who received adj CTX had a median PFS of 7.6 mos (3-12) and 4 tamoxifen (tam) failures a median PFS of 11 mos (8 15). Eleven patients who received adj CTX+tam had a median PFS of 8.5 mos (2-94). Six patients received tam at adj failure and MFL at progression. These six had a median PFS of 19.8 mos (8-50). The patients (six, who received no prior adj Tx) had a median PFS of 24.3 (8-70). MFL is as effective in achieving clinical remissions in metastatic breast cancer, is inexpensive and is far less toxic than other CTX regimes. MFL should strongly be considered as first line Tx. PMID- 11852996 TI - Order and disorder: the role of extracellular matrix in epithelial cancer. PMID- 11852999 TI - Phase I study of vinorelbine and docetaxel with granulocyte colony-stimulating factor support in the treatment of metastatic breast cancer. AB - PURPOSE: Vinorelbine and docetaxel are two active agents in the treatment of metastatic breast cancer. When given together, these drugs exhibit synergistic antitumor activity without significant pharmacokinetic interaction. The dose limiting toxicities of this combination are neutropenic fever and mucositis. Adding granulocyte colony-stimulating factor (G-CSF) might lessen the toxicity and increase the maximum tolerated dose (MTD) of this combination. The aim of this study was to determine the MTD of vinorelbine and docetaxel given in combination with G-CSF. PATIENTS AND METHODS: Between August 1997 and December 1998, 14 patients with metastatic breast cancer were enrolled in this study. All patients had received doxorubicin-based therapy, and 46% had received paclitaxel in the adjuvant or neoadjuvant setting. Patients were treated with vinorelbine at a starting dose of 20 mg/m2 intravenously over 10 min on days 1 and 5 and docetaxel at a starting dose of 85 mg/m2 intravenously over 1 hr on day 1, following the vinorelbine. Treatments were repeated every 21 days. Prophylactic G CSF 5 mcg/kg was given subcutaneously on days 3-10. Toxicity was graded according to the National Cancer Institute's grading system. RESULTS: A total of 65 cycles was administered at dose levels 0, -1, -2, and -3. The median absolute granulocyte count nadir for all courses was 200 mm(-3) (range, 0.1-7700 mm(-3)), and the median time to this nadir was 9 days (range, 7-30). The median platelet nadir was 163 (range, 27-401 k), and the median time to this nadir was 8 days (range, 7-30). The most common grade 3 nonhematologic toxicities for all courses were fatigue and myalgia, which occurred in 32 and 10 cycles, respectively. Neutropenic fever was encountered in 11 cycles. Three patients developed colitis like pictures, two of whom died as a result. Consequently, the protocol was closed to accrual before a MTD was reached. CONCLUSION: The combination of vinorelbine, docetaxel, and G-CSF in our hands has proven to be a toxic regimen, even when relatively low doses of vinorelbine and docetaxel are given. Meticulous observation of patients receiving this combination is warranted since the combination resulted in two deaths in this study. PMID- 11853000 TI - Incidence and severity of hand-foot syndrome in colorectal cancer patients treated with capecitabine: a single-institution experience. AB - BACKGROUND: Hand-foot syndrome (HFS) has been previously reported as a side effect in 45-56% of patients treated with capecitabine. However, the natural history of HFS has not been fully characterized. In this study, we investigate the incidence, severity, and time course of HFS. DESIGN: Toxicity data collected for 41 patients treated in two clinical trials were analyzedfor the occurrence of HFS. Proportions of patients developing HFS after capecitabine treatment were calculated, and the severity and time course of HFS were analyzed. RESULTS: Twenty-eight (68.3%) of the 41 patients had at least one episode of HFS. Most patients had their first (92.9%) or most severe (67.9%) episode of HFS within the first two cycles of treatment. Eighty-nine percent of the HFS episodes were grade 1 or 2; only 11% were grade 3. CONCLUSIONS: HFS is common in patients treated with capecitabine, and usually starts within the first two cycles of therapy. PMID- 11853001 TI - Repression of telomerase activity during in vitro differentiation of osteosarcoma cells. AB - In this report, we studied the relationship between telomerase activity and in vitro differentiation of osteosarcoma cells. Human osteosarcoma cells (HOS-8603) were treated with all-trans-retinoic acid (RA) and dexamethasone (DEX). Cell cycle phase, alkaline phosphatase (AP) activity, telomerase activity, and human telomerase RNA (hTR) in treated cells were detected. The results showed that the treated cells underwent morphologic differentiation. AP activity of the cells increased significantly. The proportion of the cells in S and G2/M phases was increased. A pronounced decline in telomerase activity was observed, but no significant difference in the amount of hTR expressed, when compared with the control. This study demonstrates that: (1) both RA and DEX can inhibit cell growth and induce morphologic and functional differentiation of HOS-8603 cells; (2) telomerase is an enzyme system regulated during induced differentiation of HOS-8603 cells; (3) significantly decreased telomerase activity may be an indicator of differentiation but does not parallel the expression level of hTR; and (4) the regulation of telomerase is directly linked to cell differentiation not cell cycle. PMID- 11853002 TI - Inhibitory effect of extracts of Brazilian marine algae on human T-cell lymphotropic virus type 1 (HTLV-1)-induced syncytium formation in vitro. AB - Extracts from four species of Brazilian marine algae collected from the Rio de Janeiro State coast were screened to determine the inhibitory effect on HTLV-1 induced syncytium formation. Before performing the syncytium inhibition assay the 50% cytotoxic dose (CyD50) of the algal extracts was evaluated. The antiviral test was carried out in HeLa cells co-cultured with HTLV-I infected T-cell line (C91/PL cells) in the presence of marine algal extracts in the concentration inferior to that corresponding to the CyD50. It was observed that co-cultured cells exposed to Ulva fasciata extract showed 60.2% syncytium inhibition at a concentration of 2.5%. At 5% concentration, Sargassum vulgare and Vidalia obtusiloba extracts presented 78.8 and 76% syncytium inhibition, respectively. The best inhibitory activity was observed with Laminaria abyssalis that presented 100% syncytium inhibition at a concentration of 2.5%. This work shows that extracts of marine algae, mainly L. abyssalis extract, are able to inhibit the cell-to-cell contact essential for the spreading of the virus and could be useful to prevent the infection. PMID- 11853003 TI - The concentration of serum transforming growth factor beta-1 (TGF-beta1) is decreased in cervical carcinoma patients. AB - Transforming growth factor beta-1 (TGF-beta1) is a multifunctional growth factor and known to inhibit the proliferation of epithelial cell. The relationship between serum TGF-beta1 level and the presence of cervical cancer was investigated in this study. The patients with cervical squamous cell carcinoma (SCC) had significantly lower level of serum TGF-beta1 (39.14 +/- 9.03 ng/mL; mean +/- SD) than those with myoma (49.17 +/- 9.38 ng/mL) and normal subjects (49.13 +/- 8.81 ng/mL), (p < 0.007 and p < 0.001, respectively). TGF-beta1 level was also lower in the patients with cervical intraepithelial neoplasia (CIN3) (42.07 +/- 9.38 ng/mL) than in normal controls (49.13 +/- 8.81 ng/mL) (p < 0.04). It suggested that diminution of the production of TGF-beta1 has close association with the neoplastic transformation of cervical epithelium. PMID- 11853004 TI - Randomized phase II study of irinotecan plus mitomycin C vs. oxaliplatin plus mitomycin C in patients with advanced fluoropyrimidine/leucovorin-pretreated colorectal cancer. AB - INTRODUCTION: Irinotecan and oxaliplatin are two new agents with promising activity in advanced colorectal cancer. Based on preclinical and clinical evidence that both drugs might act synergistically with mitomycin C, a randomized study using a 'pick the winner' design was undertaken to determine the effectiveness and tolerance of these two combination schedules in patients with fluoropyrimidine/leucovorin-pretreated advanced colorectal cancer. PATIENTS AND METHODS: Sixty-four patients with metastatic colorectal cancer, who progressed while receiving or within 6 months after discontinuing palliative chemotherapy with fluoropyrimidines/leucovorin were enrolled onto this study. They were randomly assigned to treatment with irinotecan 120 mg/m2 on days 1 + 15 plus mitomycin C 8 mg/m2 on day 1 (arm A) or oxaliplatin 85 mg/M2 on days 1 + 15 plus mitomycin C 8 mg/m2 on day 1 (arm B). In both treatment arms, courses were repeated every 4 weeks. RESULTS: The objective response rate in arm A is 7/33 (21.2%; 95% confidence interval, 9.0-38.9%) as compared to 5/31 in arm B (16.1%; 95% CI, 5.5-34.7%). Stable disease was noted in 48.5 vs. 45.2%, whereas the tumor progressed in 30.3 vs. 38.7%, respectively. Similar to the recorded response activities, the difference of the two combination regimens in terms of median time to progression (7.0 vs. 5.2 months) and overall survival (12.0 vs. 11.2 months) was only minor and clincally insignificant. The tolerance of treatment was acceptable in both arms, though severe adverse reactions requiring dose reductions (30 vs. 16%) and treatment delays (22 vs. 13% of courses) were more commonly noted with irinotecan/mitomycin C. The most common toxicities in arm A were neutropenia (85%; WHO grade 3/4 in 33%), thrombocytopenia (52%), diarrhea (45%), emesis (52%) and alopecia (92%). In arm B, common toxicities included neutropenia (68%; grade 3/4 in 13%), thrombocytopenia (81%), emesis (52%), and peripheral neutropathy (48%). CONCLUSIONS: Both mitomycin C combination regimens seem to provide an acceptable therapeutic index in patients with fluoropyrimidine/leucovorin-pretreated metastatic colorectal cancer. In view of the increasing need for a broader chemotherapeutic armentarium for second-line therapy of this common malignant disease, both regimens may be worthwhile to undergo further clinical investigation. PMID- 11853005 TI - Disruption of oncogene/tumor suppressor networks during human carcinogenesis. AB - Oncogenes were initially discovered as retrovirally transmitted tumor causing agents. The realization that such retroviral oncogenes constitute specifically altered versions of cellular genes-proto-oncogenes, was a landmark discovery that set the stage for the molecular and mechanistic era of cancer research. Moreover, the studies on oncogene functions have been instrumental in delineating many of the paradigms of cellular signal transduction. In contrast to the original studies in animals, oncogenic activation through retroviral transmission does not appear to be a major factor in human tumorigenesis. However, oncogenes are frequently activated by gain of function mutations or fusions with other genes, or they are aberrantly expressed due to amplification, increased promoter activity, or protein stabilization, and hence they play integral roles in the genesis of human tumors. PMID- 11853006 TI - Extension of nurse prescribing--is there a case for nurse prescribing in critical care? PMID- 11853007 TI - Nicotine replacement therapy for smokers admitted to intensive care. AB - People who smoke are addicted to nicotine. When smokers are admitted to intensive care units (ICUs) they have to cope with the effects of enforced nicotine withdrawal, as well as the fact that they are critically ill. The effects of nicotine withdrawal in smokers admitted to ICU are not well understood. Does nicotine replacement therapy (NRT) have a place in treating these symptoms and reducing the length of stay in ICU? This paper discusses the problems associated with nicotine withdrawal and the treatments available for ICU patients who smoke. PMID- 11853008 TI - Cost analysis and estimation of thoracic surgical patients with lung cancer in Greece: the case of Sotiria ICU. AB - This study aimed to estimate the hospitalization cost of thoracic surgical patients with lung cancer, treated in the intensive care unit (ICU) of Sotiria Hospital in Athens. This is the largest specialist thoracic disease hospital in the greater region of Central and Southern Greece. Methodology was based on the prospective study of patients' medical records, from which clinical and economic data were derived for a six-month period. The study sample consisted of 95 thoracic surgical patients. Economic analysis and cost estimation were based on the differentiation between nominal and real prices, occurring in the public hospital sector in Greece. Real ICU cost per patient is estimated at US$ 2,011 in 1998, while the total patients hospitalization cost was US$ 6,958. The patient's ICU cost represents about 29% of his total hospitalization cost, while the price deviation in the amounts covered by social insurance compared to the real hospitalization cost is also confirmed. It is believed that the use of economic analysis techniques could substantially contribute to the assessment of resource consumption, initially at a microeconomic and later at a macroeconomic level, and thus to the adoption of prospective payment systems, as these are encountered on an international level. PMID- 11853009 TI - Nursing the ARDS patient in the prone position: the experience of qualified ICU nurses. AB - The revival of the use of the prone position as a treatment for acute respiratory distress syndrome (ARDS) has been well documented in the medical literature, but there is little information regarding the difficulties of nursing patients in this position. The purpose of this study was to increase the body of knowledge by exploring the experiences of nurses who had cared for a patient in the prone position. A questionnaire was sent to all registered nurses in four large intensive care units (ICUs) to determine the main areas of concern when nursing patients in the prone position. There was a 62% response rate. Following this, a sample of 12 nurses volunteered to participate in three focus group interviews to further explore the issues. The main difficulties experienced related to the manoeuvre, including the timing of the move, the number of personnel and the co ordination required. Problems experienced in providing nursing care related to pressure areas, suctioning, accidental injuries and management of emergencies. Deficits in knowledge of ARDS and skills in handling communication with relatives were also identified. As a result of this exploration, guidelines have been developed, focusing mainly on the manoeuvre, organizational and nursing issues, to provide guidance in caring for a patient when being nursed in the prone position. PMID- 11853010 TI - Staff empowerment in Finnish intensive care units. AB - The purpose of this study was to describe staff empowerment in Finnish intensive care units. The data were collected with a questionnaire comprising demographic background and empowerment items. The concept of empowerment was divided into three components: behavioural, verbal and outcome empowerment. The questionnaire was sent to all registered nurses at Finnish intensive care units (ICUs). Eight hundred and fourteen replied, giving a response rate of 77%. The ICU nurses demonstrated confidence in their own skills and competencies, although least so in the domain of outcome empowerment. Experience of behavioural, verbal and outcome empowerment increased linearly with age. The length of nursing experience was positively associated with behavioural, verbal and outcome empowerment. Experience in ICU nursing correlated positively with verbal and outcome empowerment. Motivation, job satisfaction, respect of job autonomy and the fact that the job of ICU nurses commanded respect in society were associated with behavioural, verbal, and outcome empowerment. PMID- 11853011 TI - Factors influencing the turnover of intensive care nurses. AB - Access to the intensive care unit (ICU) is often tortuous as there is a high incidence of bed closure due to staff shortage, a problem exacerbated by a high rate of turnover. It is proposed that the first step in addressing the problem is to explore the reasons why people leave, illuminating areas of policy and practice that would benefit from amendment. Given that the issues concerned are complex and contextual in nature, an open exploratory approach was adopted, whereby respondents were interviewed using open questions and given the freedom to shape their responses according to their perceptions of the problem. The transcripts of eleven interviews were analysed, with four themes emerging which represent the most dominant factors influencing the respondents' decision to leave. These included stresses related to the work; inadequate opportunity for professional development; recognition and respect of others and the implications of shift-work. It was concluded that there needs to be a greater awareness of the potential for nurses to become excessively stressed in the ICU environment; that a decentralized management style may help maintain motivation; that rostering systems need to retain flexibility; and that there is a requirement for greater incentives to pursue a career in intensive care. PMID- 11853012 TI - Level of dyspnoea experienced in mechanically ventilated adults with and without saline instillation prior to endotracheal suctioning. AB - The purpose of this study was to compare the level of dyspnoea with and without the use of 5-cc saline instillation prior to endotracheal suctioning of mechanically ventilated adults. A crossover, quasi-experimental design was used. Seventeen alert, mechanically ventilated adults were asked to rank their level of dyspnoea using the vertical visual analogue scale at specific time intervals surrounding two suctioning events. Saline was randomly assigned to be instilled prior to one of two suctioning episodes. Dyspnoea was ranked immediately after suctioning, and at 10-, 20-, and 30-minute intervals. Data were analyzed using repeated measures analysis of variance with time of measure (immediately after suctioning, 10-, 20-, and 30-minute intervals) and treatment type (with saline versus without saline instillation). The level of dyspnoea based on treatment type (with or without saline) was non-significant. Treatment type by age group interaction was significant (F(1, 15) = 5.41, P = 0.034). The nature of the interaction revealed that older patients (< or = 60 years of age) experienced less dyspnoea without saline prior to suctioning and greater dyspnoea with saline instillation as compared to the younger subjects (<60 years of age). This study documented no beneficial effects of saline. However, it did demonstrate that saline instillation might precipitate a significantly increased level of dyspnoea for up to 10 minutes after suctioning in patients older than 60 years of age. Recommendations based on the results of this study would be to avoid the use of saline instillation prior to suctioning. PMID- 11853013 TI - Clopidogrel--a myocardial infarction CURE? AB - Recently the anti-platelet drug clopidogrel has been publicized in the press as a breakthrough in the treatment of myocardial infarction. PMID- 11853014 TI - Chest X-ray quiz. Bronchopneumonia. PMID- 11853015 TI - Fifty ways to make a neuron: shifts in stem cell hierarchy and their implications for neuropathology and CNS repair. AB - During embryogenesis, the developmental potential of individual cells is continuously restricted. While embryonic stem (ES) cells derived from the inner cell mass of the blastocyst can give rise to all tissues and cell types, their progeny segregates into a multitude of tissue-specific stem and progenitor cells. Following organogenesis, a pool of resident "adult" stem cells is maintained in many tissues. In this hierarchical concept, transition through defined intermediate stages of decreasing potentiality is regarded as prerequisite for the generation of a somatic cell type. Several recent findings have challenged this view. First, adult stem cells have been shown to adopt properties of pluripotent cells and contribute cells to a variety of tissues. Second, a direct transition from a pluripotent ES cell to a defined somatic phenotype has been postulated for the neural lineage. Finally, nuclear transplantation has revealed that the transcriptional machinery associated with a distinct somatic cell fate can be reprogrammed to totipotency. The possibility to bypass developmental hierarchies in stem cell differentiation opens new avenues for the study of nervous system development, disease, and repair. PMID- 11853016 TI - Expression of nicotinamide N-methyltransferase (E.C. 2.1.1.1) in the Parkinsonian brain. AB - Nicotinamide N-methyltransferase (NNMT) has been proposed as a link between the environmental and genetic factors of Parkinson disease (PD). Therefore, we explored the hypothesis that high levels of NNMT expression may predispose to the development of PD. Regions of high mRNA expression were shown in the spinal cord, medulla, and temporal lobe, with lowest expression in the cerebellum, subthalamic nucleus, and caudate nucleus. Using 2 NNMT antibodies, the protein was shown to be expressed in multipolar neurons in the temporal lobe, caudate nucleus, and spinal cord, granular neurons of the cerebellum, dopaminergic neurons in the substantia nigra, and in the axons of the third nerve. Expression of NNMT was compared in PD and non-PD control cerebella and caudate nucleus. PD tissue exhibited significantly increased levels of NNMT protein and activity. PD disease duration was inversely correlated with the level of expression in cerebellum. This is the first demonstration that patients with PD have higher levels of NNMT activity and protein in brain tissue than those without PD and that NNMT expression is associated with neurons that degenerate in PD. PMID- 11853017 TI - Effects of brain-derived neurotrophic factor (BDNF) on compression-induced spinal cord injury: BDNF attenuates down-regulation of superoxide dismutase expression and promotes up-regulation of myelin basic protein expression. AB - Neurotrophins enhance the survival of cells in the nervous system under both physiological and pathological conditions, such as those caused by disease or trauma. We recently demonstrated that expression of brain-derived neurotrophic factor (BDNF) was up-regulated in neurons and glia after compression-induced spinal cord injury (SCI). We show here the effects of BDNF on the oligodendrocyte survival and functional recovery after SCI. The effects of intrathecally administered BDNF on both Cu/Zn superoxide dismutase (CuZnSOD) and myelin basic protein (MBP) expression were examined using rats that had received compression induced spinal cord injury. CuZnSOD expression in the spinal cord was down regulated within 24 h of compression-induced injury and then recovered. Continuous infusion of BDNF inhibited the acute down-regulation of CuZnSOD expression. In situ hybridization showed that CuZnSOD was expressed in both neurons and glia. Although MBP expression was greatly reduced after injury, BDNF administration promoted the recovery of MBP expression nearly to a control level after 2 wk. Furthermore, BDNF administration also prompted behavioral recovery. These results suggest BDNF's usefulness in human clinical applications. The attenuation of CuZnSOD down-regulation may be related to a protective effect of BDNF and the promotion of MBP up-regulation may be related to a long-lasting restorative effect. PMID- 11853018 TI - Evidence for separable functions of tuberous sclerosis gene products in mammalian cell cycle regulation. AB - Tuberous sclerosis is an autosomal dominant disease affecting approximately 1 in 6,000 individuals. It is caused by mutations in either TSC1 on chromosome 9q34, which encodes hamartin, or TSC2 on chromosome 16p13.3, which encodes tuberin. The growths, named hamartomas, characteristically occur in different organs of patients and are speculated to result from defects in proliferation control. The observation that hamartin and tuberin can interact in vivo suggests that they might function in the same complex. Here we show that hamartin can affect proliferation control independent of the presence of functional tuberin and that binding to hamartin is not essential for tuberin to affect proliferation. Ectopic expression of hamartin negatively regulates proliferation to a similar extent in tuberin-positive and tuberin-negative cells; this is accompanied by binding to tuberin and upregulation of endogenous p27 in tuberin-positive cells and is without effects on p27 expression in the latter. Our data show for the first time that TSC proteins possess separable functions. We further demonstrate that hamartin can deregulate proliferation control by different mechanisms depending on the presence of tuberin. Besides an overlap in many features of patients with TSC1 and TSC2 mutations, data has accumulated that provides evidence for specific clinical differences. This study provides new insights into the cellular roles of TSC proteins and initiates a discussion of whether separable functions of these proteins might be associated with the clinical differences of TSC1- and TSC2 associated disease. PMID- 11853019 TI - Decreased expression of hippocampal cholinergic neurostimulating peptide precursor protein mRNA in the hippocampus in Alzheimer disease. AB - Hippocampal cholinergic neurostimulating peptide (HCNP) is involved in the phenotype development of the septo-hippocampal system. HCNP precursor protein (HCNP-pp) is known to interact with other molecules including phosphatidylethanolamine and Raf-1 kinase, and is also known as phosphatidylethanolamine-binding protein and raf kinase-inhibitory protein. To assess whether HCNP-pp is involved in the pathogenesis of Alzheimer disease (AD), the expression levels of its mRNA in the hippocampus of autopsy brains from patients with dementia (including AD and ischemic vascular dementia) were compared with those of non-demented control subjects. The in situ hybridization analysis revealed that the expression of HCNP-pp mRNA in patients with clinically late-onset AD was decreased in the hippocampal CA1 field, but not in the CA3 field or the dentate gyrus. The early-onset AD patients showed a wide range of expression levels in the hippocampal sub-regions. Northern blot analysis of HCNP pp mRNA in brain tissue supported these observations. Since HCNP is known to stimulate the enzymatic activity of choline acetyltransferase in neurons, its low expression in the CAI field of AD patients may explain the downregulation of cholinergic neurons seen in these patients and may thus contribute to the pathogenic processes underlying AD. PMID- 11853020 TI - Increase of preproenkephalin mRNA levels in the putamen of Parkinson disease patients with levodopa-induced dyskinesias. AB - The expression of preproenkephalin messenger RNA was studied in the brain of Parkinson disease (PD) patients using in situ hybridization. All these patients were treated with levodopa (LD) and the development of motor complications was recorded. Eleven normal controls and 14 PD patients were used, of which 4 developed dyskinesias, 3 developed wearing-off, 3 developed both dyskinesias and wearing-off, and 4 developed no adverse effect following dopaminomimetic therapy. Nigrostriatal denervation was similar between the subgroups of PD patients as assessed using 125I-RTI-specific binding to the dopamine transporter and measures of catecholamine concentrations by HPLC. A significant increase of preproenkephalin messenger RNA levels was observed in the lateral putamen of dyskinetic patients in comparison to controls (+210%; p < 0.01) and in comparison to nondyskinetic patients (+112%; p < 0.05). No change was observed in medial parts of the putamen or in the caudate nucleus. No relationship between preproenkephalin messenger RNA levels and other clinical variables such as development of wearing-off, age of death, duration of disease, or duration of LD therapy was found. These findings suggest that increase synthesis of preproenkephalin in the medium spiny output neurons of the striatopallidal pathway play a role in the development of dyskinesias following long-term LD therapy in Parkinson disease. PMID- 11853022 TI - Hypomagnesemia in acute and chronic illness. AB - Magnesium deficit is associated with several acute and chronic illnesses. Of major concern is the association between cardiovascular problems, such as myocardial infarction, hypertension, congestive heart failure, and hypomagnesemia. In addition, evidence is mounting regarding the relationship between Type II Diabetes Mellitus, and magnesium deficit. The American diet is low in magnesium, and with modern water systems, very little is ingested in the drinking water. A review of the state of the science in relation to literature on magnesium follows, as well as nursing interventions crucial to managing magnesium deficit. PMID- 11853021 TI - Arrested oligodendrocyte lineage progression during human cerebral white matter development: dissociation between the timing of progenitor differentiation and myelinogenesis. AB - Immature oligodendrocytes (OLs) derive from a large pool of late OL progenitors that populate human cerebral white matter throughout the latter half of gestation. We recently reported that a minor population of immature OLs are present in human cerebral white matter for at least 3 months before these cells commit to myelinogenesis around 30 wk postconceptional age. Since this finding supports dissociation between the events that regulate human immature OL maturation and their commitment to myelinogenesis, we characterized here the cellular sequence of events that characterize immature OLs during the transition from a premyelinating to a myelinating state. Commitment of immature OLs to myelinogenesis in human cerebral white matter correlated with the longitudinal extension of specialized processes, designated "pioneer processes," that made multiple types of apparent contacts with axons. This event coincided with the appearance of 3 distinct populations of sheaths that varied in their labeling for myelin basic protein (MBP). Since few axons initially labeled for MBP, this supported the occurrence in vivo of O4-negative, O1-positive premyelin sheaths that precede MBP-positive compacted myelin. These observations identify 3 sequential stages of early myelinogenesis: 1) the initial ensheathment of axons by premyelin sheaths generated by immature OLs; 2) the initial insertion of MBP into transitional sheaths; and 3) the generation of MBP-rich mature myelin. PMID- 11853023 TI - Elder abuse: critical care nurse role in detection. AB - With an increasing number of elders entering hospital critical care units, the nurse in this setting has a responsibility for detecting signs and symptoms of elder abuse. Elder abuse is manifest in several ways including inflicted physical harm as a result of traumatic injuries or neglect, psychologic threats or intimidation, and even financial abuse when the elder's resources have been misappropriated by the caregiver. An Assessment Guide for Elder Abuse is an excellent screening tool for critical care nurses in detecting this escalating problem. PMID- 11853024 TI - Recognition and documentation of domestic violence in the clinical setting. AB - Critical care nurses may encounter a victim of domestic violence or abuse when caring for an individual with traumatic injuries. Understanding the injuries that are associated with acts of violence is only the first step. A vital part of the nurse's responsibilities is the precise written documentation of observations, physical assessments, and other factors that may later become vital evidence in a court of law. PMID- 11853025 TI - Early recognition of Munchausen Syndrome by Proxy. AB - In Munchausen Syndrome by Proxy (MSBP), a caregiver fabricates or induces illness in another individual. In most cases, the parent is the perpetrator and the child, the victim. MSBP is a form of child abuse that in extreme cases may result in death. A large percentage of health care providers are not familiar with the syndrome. Early recognition of MSBP is necessary to reduce morbidity and mortality associated with the syndrome. PMID- 11853026 TI - Shaken baby syndrome. AB - "Shaken baby syndrome" (SBS) results in intracranial and intraocular hemorrhages with no evidence of external trauma. The cause of these injuries is vigorous shaking of an infant being held by the chest, shoulders, or extremities. A significant number of cases of head trauma in infants and young children are a direct result of SBS. In extreme cases, SBS may result in death. Many cases of SBS go unidentified because of the absence of external injuries, no witnesses to the event, and the failure of the abuser to admit his or her actions. Because of the numbers of children affected and the difficulty in detecting SBS, it is essential that critical care nurses who care for children be aware of SBS as a potentially lethal form of abuse. This article educates nurses about the signs and symptoms of SBS in an attempt to foster early recognition and interventions in cases of SBS to reduce mortality and morbidity associated with the syndrome. PMID- 11853027 TI - Health care providers' perspectives on breaking bad news to patients. AB - This article reports the results of an investigation designed to obtain descriptive information about what typically transpires in bad news transactions between patients and physicians. A sample of 115 health care providers who were attending a 1-day workshop on palliative care issues responded to questions regarding bad news transactions between physicians and patients. Results indicated that giving the news in person, giving the news in a private place, having patient support providers present, and using a warm and caring tone are highly typical of bad news transactions, whereas exploring patient emotional reactions, relying on touch, delivering the news at the patient's pace, and providing written information are less typical. Nurses and physicians diverged in the perceptions about what typically transpires, suggesting that studies focusing only on physician reports or recommendations may be misleading. These data also point to the need to obtain other views of bad news transactions, and they argue for research designed to assess the relation between actual patient-physician encounters and subsequent patient-related outcomes. PMID- 11853028 TI - Critical care nurses' perceptions of family needs as met. AB - In this descriptive, exploratory study, nurses' perceptions of family needs as met during the critical care experiences of an adult member were correlated to the families' perception of those same needs as being met. The population consisted of 45 family members in a large county hospital designated as a Level 3 trauma center. Family members of adult patients and registered nurses who were assigned to care for them completed a three-part instrument, which consisted of the Demographics Data Questionnaire, the Critical Care Family Needs Inventory (CCFNI), and the Needs Met Inventory (NMI). Data were analyzed using descriptive and inferential statistics. The top 10 needs perceived by the family members and registered nurses were reported in order of importance during the first 18-24 hours on the CCFNI and NMI. Data were analyzed on all 45 need statements to determine the top 10 needs perceived as important on the CCFNI and perceived as being met on the NMI. A one-way analysis of variance (ANOVA) test was performed on the data and yielded significant differences on three of the items. Linear regression was performed using t test which supported a significant difference on five statements based on critical care nursing years of experiences in critical care. Self-reported or open-ended comments from the family members and nurses were presented. PMID- 11853029 TI - Endothelin-mediated preeclampsia at high altitude. AB - Preeclampsia is pregnancy-induced hypertension. The hypoxia at high altitude increases the incidence of preeclampsia. Endothelin is released in response to hypoxia and is associated with other hypertensive states at high altitude. Endothelin may play a major role in preeclampsia for individuals residing at high altitude. Endothelin antagonists may prove useful in the treatment of preeclampsia. PMID- 11853030 TI - Noninvasive blood gas monitoring. AB - In the clinical setting, noninvasive blood gas monitoring has become the standard of care over arterial punctures. Technology has provided ways to measure both arterial oxygen (PaO2) and arterial Carbon Dioxide (PaCO2). With the availability of noninvasive blood gas monitoring, patient care and comfort is improving, and cost savings are being implemented. Overall noninvasive monitoring can aid in the diagnosis of some pulmonary diseases and monitor patient progress. PMID- 11853031 TI - Women in physical medicine and rehabilitation. Leadership issues. AB - Academic medical centers share the triple missions of education, research, and clinical care, particularly in vulnerable populations. They are well positioned to have a significant impact on reducing health disparities between patients from diverse backgrounds, both now and in future generations. A key component to realizing this goal lies in increasing the number of women in academic medicine. Women bring a fresh perspective to the investigative process, often targeting gender is a better way of providing preventive services, and patient satisfaction studies have shown that many patients prefer women physicians. PMID- 11853032 TI - Osteoporosis in women with spinal cord injuries. AB - Decreased bone density and increased fracture risk are seen in patients with SCI. The bone resorption rate is markedly increased. Hypercalciuria, low PTH, and low 1,25 (OH)2 vitamin D are commonly seen. Bed-rest studies show similar findings, but of lesser magnitude. Therapies to treat or prevent osteoporosis include optimal nutrition (with care to avoid exacerbating hypercalciuria). Weight bearing or functional electrical stimulation cycle ergometry may prevent some of the bone loss, especially in acutely injured patients. Estrogen should be considered in postmenopausal or amenorrheic women, but not if they are at high risk of thromboembolism. More research on effects of estrogen is needed in this population. Bisphosphonates may also help prevent the acute bone loss; oral routes must not be used in recumbent patients. Thiazides could be useful as adjunct therapy. PMID- 11853033 TI - Menopause and aging with disability. AB - Aging involves change and adaptation to change. The normal or usual changes of aging often have significantly greater impact on an individual whose disability has limited his or her physical or socioeconomic reserves. The aging process itself may be accelerated by overuse and compensatory mechanisms. The changes of aging have unique features in damaged body systems that exhibit physiologic adaptations. The changes of aging therefore often result in secondary impairments, leading to secondary disability. New adaptations or repeated rehabilitation are needed to regain the equilibrium among biologic, psychosocial, and environmental influences. Disabled women do not seem to have a significantly different experience of menopause from other women, but postmenopausal changes accelerated bone loss and increased risk of heart disease-do appear to carry greater risk in those with mobility impairment. Hormone replacement therapy has both greater potential benefit and greater potential risks. Review of these issues makes evident the great need for research in the area of aging with disability, improvement in physician and consumer education, and future health care planning. PMID- 11853034 TI - The health of women with cerebral palsy. AB - Over the past 10 years, there has been interest in describing the health and functional status of persons with CP. More recently, information specific to women with CP has become available. From these studies a better appreciation of the affect a lifelong primary disability such as CP has on an individual's life has developed. An understanding of aging with a disability, secondary conditions, associated conditions, comorbidities, and health have promoted a health and wellness agenda for women and men with CP. Women with CP are generally healthy. There appears to be no significant risk for any specific comorbiditv from CP. Women may note a modest change in function over time that may represent issues of aging, but there should be no dramatic loss of function. Any loss of function must be evaluated further for cause, and cannot be attributed to aging alone. Common secondary conditions and health issues include pain and musculoskeletal issues, bladder and bowel problems, poor dental hygiene, and possibly gastroesophageal reflux. Osteoporosis at all ages from limited mobility (secondary osteoporosis) likely is common, but only recently has bone densitometry been used for more routine evaluations; consequently, no data are available in women with CP regarding this. All health issues should be evaluated and appropriate intervention prescribed. There appear to be no significant reproductive health issues for women with CF. Women should engage in all decisions regarding diagnosis and treatment. Women with CP likely participate in some healthy behaviors. However implementing health promotion programs requires some understanding of health behavior theories and models. It may not be enough to modify existing programs for accessibility (e.g., transportation, cost. environment). Engagement in health promotion programs by women with CP must take into account individual perceptions and values, social networks, a sense of personal control, and a readiness to change attitude toward changing lifestyles. PMID- 11853035 TI - Women and traumatic brain injury. AB - Women with TBI have been inadequately studied in relation to most aspects of pathophysiology, recovery, health and behavioral issues, and community integration. This is not entirely surprising in light of the preponderance of men with TBI but also reflects the traditional tendency of medical researchers to concentrate their efforts on men. Although most of the residual effects of TBI are gender-neutral, women may present some unique problems in relation to pain and endocrine issues, reproduction, and sexual functioning In addition, a woman's roles as wife, mother, and daughter are likely to result in a different constellation of family dynamics when TBI is introduced. Attention to enrollment of women in research studies and the increasing number of multi-institutional studies of TBI may provide enlightenment on these issues in the future. PMID- 11853036 TI - Gynecologic and reproductive issues in women with spinal cord injury. AB - As the treatment of SCI and its secondary medical complications improves, women with SCI are becoming increasingly successful in re-integrating themselves into society. Part of this re-integration involves the resumption of active sexual lives and motherhood. Clinicians who care for patients with SCIs play an important role in facilitating the resumption of these roles. With appropriate follow-up and care, women with SCI should be expected to maintain good gynecologic health and deliver healthy children with minimal complications. PMID- 11853037 TI - Quality of life for women with physical disabilities. AB - This article emphasizes the role of several factors on the QOL of women with disabilities. Future studies might examine ways to merge these factors to examine their long-term consequences in the lives of women with disabilities and, hence, society at large. Findings should provide a basis for policymaking and clinical rehabilitation interventions designed to promote greater QOL for women with disabilities-a QOL that offers vigorous prosperity, flourishing health, and full social participation. PMID- 11853038 TI - Physical activity and exercise for women with mobility impairments. AB - Decisions regarding exercise or physical activity for women with mobility impairments are more complex than those made by other women and by people without mobility limitations. The epidemiologic caveat that being active improves health and well-being generally is applicable, but on the individual level there will be exceptions. For many mobility-limiting conditions, there has been a paucity of research that includes women and even less that focuses specifically on women. Although there is beginning to be evidence that this is changing, there are many unaddressed needs and opportunities for future study. PMID- 11853039 TI - Sexual function in women with neurologic disorders. AB - Significant gaps remain in our understanding of the impact of most neurologic disorders, including brain dysfunction and MS. Recent methodology used to assess women with SCIs may be used as a model with which to study these other disorders. The availability of oral agents to treat sexual dysfunction in men should also stimulate research using these agents to treat sexual dysfunction in women. PMID- 11853040 TI - Health promotion practices in women with multiple sclerosis: increasing quality and years of healthy life. AB - Disease-modifying agents that decrease the accumulation of disability are costly, inconvenient to use, and have the potential of significant side effects. The findings presented here suggest that much less costly health-promotion interventions may hold substantial promise as a complementary strategy for decreasing the burden of functional limitations. Additional research is needed to develop and test interventions that rehabilitation professionals can use to enhance physical activity, nutrition, stress management, and other health promoting behaviors to increase the quality and years of healthy life for women with MS. PMID- 11853042 TI - The effect of a church-based breast cancer screening education program on mammography rates among African-American women. AB - This study examines the effectiveness of breast cancer screening education programs on mammography rates among African-American women 40 years of age and over. We conducted two types of educational programs in community settings, primarily in African-American churches. Three-month follow-up interviews were used to determine whether women who participated in programming were more likely to get a mammogram if they had not had a mammogram in the last year. Our results demonstrate that the educational programs significantly increased the likelihood of getting a mammogram when compared to a control group that received no educational programming. Further, we found that the programs were effective for motivating breast cancer screening in housing projects as well as in the churches, and that the effectiveness of the programs remained even when we controlled for socioeconomic status, depression, and age. PMID- 11853041 TI - Bladder management in women with neurologic disabilities. AB - Women with neurologic disabilities can experience an array of bladder disorders. There are many aspects of lower urinary tract management that are specific to women. Treatments should be based on urodynamic findings, and individualized for each woman's physical and cognitive abilities. PMID- 11853043 TI - Mammography and diagnosis of breast cancer. PMID- 11853044 TI - Screening children for tuberculosis based on residence in a high-prevalence inner city community. AB - In January 1994, the Committee on Infectious Diseases of the American Academy of Pediatrics (AAP) published detailed guidelines on tuberculin skin testing of children. These guidelines required mandatory screening of all children residing in high-prevalence communities. In February 1996, the AAP published updated and revised guidelines that allowed for targeted screening of such children based on epidemiologic risk of acquiring tuberculosis. We report on an assessment of the effectiveness of mandatory screening for tuberculosis in a high prevalence community. PMID- 11853045 TI - Irritant contact dermatitis complicated by deep-seated staphylococcal infection caused by a hair relaxer. AB - Chemical hair relaxers are used by many women to straighten their hair. We describe a case of a deep soft tissue staphylococcal abscess that complicated an irritant contact dermatitis from a hair relaxer treatment. PMID- 11853046 TI - Red degeneration of a leiomyoma masquerading as retained products of conception. AB - BACKGROUND: The incidence of leiomyomas in pregnancy is approximately 1%. Their presence has been linked to spontaneous abortion, premature labor, soft tissue dystocia, uterine inertia, fetopelvic disproportion, malposition of the fetus, retention of the placenta, and postpartum hemorrhage. CASE: This case report documents a seldom-described event of a submucous leiomyoma masquerading as retained products of conception. The patient presented 4 weeks postpartum with complaints of urinary retention and heavy bleeding with cramping. Examination revealed a large mass resembling placental tissue filling the vaginal vault. The necrotic mass was removed with blunt and sharp dissection. The final pathology report revealed a degenerating leiomyoma. CONCLUSION: Complicating factors associated with this fibroid included a history of spontaneous abortion and preterm labor, as well as fetal malpresentation and carneous degeneration of the leiomyoma. Gestational myomas, although rare, can have an unusual appearance that may be misinterpreted. PMID- 11853047 TI - Overcoming the barrier of lactose intolerance to reduce health disparities. AB - Federal health goals for the public have focused on reducing health disparities that exist between whites and various racial and ethnic groups. Many of the chronic diseases for which African Americans are at greater risk- hypertension, stroke, colon cancer, and obesity-may be exacerbated by a low intake of calcium and/or other dairy-related nutrients. For example, a low intake of dairy food nutrients, such as calcium, potassium, and magnesium, may contribute to the high risk of hypertension seen in African Americans. The Dietary Approaches to Stop Hypertension (DASH) study demonstrated that a low-fat diet rich in fruits and vegetables (8 to 10 servings) and low-fat dairy foods (3 servings) significantly reduced blood pressure-and was twice as effective in African-American participants. Calcium and dairy food consumption is particularly low among African-American, Hispanic, and Asian populations. Average intakes are near the threshold of 600 to 700 mg/day, below which bone loss and hypertension can result. Although lactose intolerance may be partly to blame for the low calcium intakes due to reduced dairy food consumption by minority populations, culturally determined food preferences and dietary practices learned early in life also play a role. The high incidence figures for primary lactose maldigestion among minority groups grossly overestimates the number who will experience intolerance symptoms after drinking a glass of milk with a meal. Randomized, double-blind, controlled clinical trials have demonstrated that by using a few simple dietary strategies, those who maldigest lactose (have low levels of the lactase enzyme) can easily tolerate a dairy-rich diet that meets calcium intake recommendations. Physicians and other health professionals can help their minority patients and the general public understand how to improve calcium nutrition by overcoming the surmountable barrier of lactose intolerance. At the same time they will be helping to reduce the incidence of calcium-related chronic diseases for which minority populations are at high risk. PMID- 11853048 TI - On any Saturday--a practical model for diabetes education. AB - PURPOSE: Patient self-management is an important part of treating chronic diseases. However, many primary care physicians face barriers in offering office based diabetes education. This paper will discuss a practical program of community-based diabetes education that can be easily modified for a practitioner's office. PROCEDURE: Half-day diabetes education workshops geared toward local health care providers and patients with diabetes and their families were conducted in two rural communities in Arkansas. Participants were surveyed with respect to the effectiveness of the program and how they would use what they learned in the program. FINDINGS: Thirty-one health care providers and 59 patients with diabetes and their families attended. Program evaluation scores were between 4.1 and 5 on a 5-point Likert scale. One third of the patients commented that they had a better understanding of diet and medication use. Feedback from community health care providers noted that attendance in local diabetes support groups increased after the workshops. CONCLUSIONS: Diabetes complications have a large impact on the health of the population and a growing economic impact on the health care industry. Although there are many barriers to diabetes education and control, a practical half-day diabetes workshop on any Saturday can be effectively developed and implemented. PMID- 11853049 TI - Iron deficiency anemia in HIV infection: immunologic and virologic response. AB - We report the immunologic and virologic response of iron deficiency anemia in two cases of human immunodeficiency virus (HIV)-infected individuals. The findings of this report suggest that caution be exercised in prescribing iron supplements to HIV-infected patients. The treatment of iron deficiency anemia should be combined with antiretroviral agents in people living with HIV/AIDS to avoid adverse immunologic and virologic consequences. PMID- 11853050 TI - Tobacco control: consensus report of the National Medical Association. AB - ISSUES: Tobacco Control remains one of the greatest determinants for reducing the morbidity and mortality of African Americans. OBJECTIVE: To examine the scope and consequences of tobacco use among African Americans and characterize its implications for the National Medical Association physician membership and their patients, and identify policy, education, advocacy and research issues in Tobacco Control for the organization. CONSENSUS PROCESS: Literature review using the MEDLINE database from January 1966 to August 1999 Week 1, searching Medical Subject Heading (MeSH) reading combined with text words "Black" or "African American" and "Tobacco" as a search term, identified 130 articles/110 abstracts published between 1988 and February 1999. The panel selected 61 appropriate articles and a paper summarizing the literature review was developed. The summary paper was used as background material for a formal consensus panel discussion on July 16-17, 1999. Consensus among committee members was reached via mail, fax and e-mail using the summary review paper, annotated bibliographies key informant surveys, and previous NMA resolutions on tobacco control. A formal working session was held on July 16-17, 1999 in which four areas of concentration of issues were determined: Policy, Advocacy, Education and Research. All committee members approved the final report. SUMMARY: Because tobacco control issues in African Americans are both complex and poorly understood, the panel views the NMA's role as pivotal in the coordination of resources and capacity-building to address all four areas identified. Stronger partner-ships with traditional federal and nonprofit agencies associated with tobacco control/advocacy in African Americans as well as nontraditional organizations (i.e., churches, academia, marketing and media organizations) also must occur to strengthen the infra-structure needed to assess needs, design appropriate interventions and evaluate the appropriateness, effectiveness and efficacy of tobacco control efforts in African American communities. PMID- 11853051 TI - Less is better. AB - The "more is better" credo is evaluated in terms of its harmful ramifications on human health, the environment, and the survival of the human race. The trend towards greater height and body weight in developed countries is evaluated in terms of its negative aspects on health and longevity. The benefits of reduced caloric intake are discussed. Countries that survive on lower food intakes are shown to have much less heart disease, such as South Africa, where rural blacks outlive whites and also have a higher percentage of centenarians. The risks of increasing birth weight are discussed in terms of promoting cancer and overweight in adulthood. Rapid childhood growth also is shown to have risks because rapid maturity is in conflict with the need to have more time to learn about our complex world before reaching reproductive capability. The increase in the average size of humans aggravates our burgeoning population numbers, placing even greater demands on our need for fresh water, energy, resources, and a clean invironment. Many good things come in large packages, such as elephants, whales, and trees. While Sumo wrestlers, football players, and basketball players play an important role in our world, their small numbers do not pose a threat to our survival. However, their body habitus should not be the goal for the average male of the future. PMID- 11853052 TI - Program to prevent pregnancy while on medication. PMID- 11853053 TI - Hand washing decreases risk of colds and flu. PMID- 11853054 TI - Building a healthy community. PMID- 11853055 TI - Wake of September 11th attacks: implications for research, policy and practice. AB - The National Consortium for African American Children (NCAAC) held a National Forum on Bioterrorism and Children on November 6, 2001 in Washington, DC. Convened in the wake of the September 11th attacks on New York City and the Pentagon, this unprecedented conference assembled a cross-section of professionals in child advocacy, health, mental health, insurance, economics, law enforcement, and media technology. The ensuing discussion focused on issues surrounding biological agents, their impact on children and youth, and the strategies needed to protect the health and mental health of children and families in the event of a large-scale bioterrorist crisis. Lessons learned as well as the implications of the terrorist acts from the tragic events of September 11th formed the backdrop for engaging dialogue among various industry executives and professionals. Accounts of personal experiences during the unprecedented tragedy of 9-11 were shared and provided a context for heightened preparedness planning for children and adults. A collaborative statement was also presented by NCAAC, the National Medical Association, the Association of Black Psychologists, and the National Black Media Coalition. This forum was hailed as a model for communities of color to join and help bolster broad-based coalition building to ensure the availability of culturally and linguistically appropriate messages, services, and support. As intended, the forum devoted significant attention to the special needs of children, their caregivers and families and provided for an invaluable interchange which is slated to evolve into a national action plan to address the imminent dangers facing our nation's children. PMID- 11853056 TI - Syphilis on the decline. PMID- 11853057 TI - Black TV shows send unhealthy food messages. PMID- 11853058 TI - My company or my career: managerial achievement and loyalty. AB - This article examines the objective and subjective aspects of managerial careers in six large firms that have experienced organizational restructuring. We begin by assessing the dominant models of change in career structures, particularly those which emphasize the 'portfolio' route to career success. Although elements of the bureaucratic career remain, we find some evidence of the shift predicted by these models amongst the younger generation of managers. However, it is striking that not all young managers are able to take advantage of the opportunities offered by the new career model. Indeed, cultural capital has an increasing impact on career achievement. Younger managers are responding by reorienting away from organizational loyalty towards a concern with individual career projects. PMID- 11853059 TI - Industrialization, class structure, and social mobility in postwar Japan. AB - This study examines intergenerational class mobility in Japan using cross national comparisons with Western nations and cross-temporal comparisons of five national surveys conducted in postwar Japan. Cross-national comparisons highlight the similarity in relative mobility pattern between Japan and Western nations and at the same time the Japanese distinctiveness in absolute mobility rates especially regarding the demographic character of the Japanese manual working class. The results of cross-temporal comparisons of mobility pattern report some systematic trends in total mobility, inflow and outflow rates, reflecting the Japanese experience of late but rapid industrialization. The pattern of association between class origin and class destination, however, was stable in postwar Japan. It is therefore the combination of distinctive absolute mobility rates and similar relative mobility rates that characterizes the Japanese mobility pattern in comparison with the Western experience. Furthermore, Japan's distinctive pattern of postwar social mobility is characterized by a combination of rapidly changing absolute mobility rates and comparatively stable relative mobility rates. PMID- 11853060 TI - Socio-political control in urban China: changes and crisis. AB - This paper examines urban China's socio-political control crisis under the impact of economic reforms as an epitome of a more general social crisis. The traditional urban institutional form of socio-political control in the People's Republic of China (PRC), the work unit form of control, is a variant of age-old forms. The latter's reproduction in variant form in the former was premised upon the fact that the PRC's industrialization was carried out by a peasant-based party creating a new working class of rural migrants engaged in non-market production and exchange. The persistence of non-market economic relations ensured this form of control's continued reproduction. Post-1978 market-oriented reforms have undermined this form. As the emergence of new forms has been slow, a socio political control crisis has arisen, at a time when millions of urban employees are being thrown out of work. In dealing with the crisis, the official trade union, an organic constituent institution of the work unit form of control, plays a prominent part, in being given the tasks of sustaining this decaying form, and preventing and defusing potential social explosion. Yet, the very economic reform programme that has undermined the work unit form of control, is also gravely weakening the union. PMID- 11853061 TI - Social mobility and constitutional and political preferences in Northern Ireland. AB - During the past thirty years Catholics in Northern Ireland have experienced unprecedented upward social mobility. Some commentators have suggested that this has led Catholics not merely to adopt the lifestyles of the middle class but also to modify their constitutional preferences, leading to a decline in nationalism. In this paper I examine the relationship between social mobility, on the one hand, and, on the other, both constitutional preferences and political (left or right wing) preferences among Catholics and Protestants in Northern Ireland, using survey data collected in 1996. There is no evidence that Catholics' constitutional preferences are related to their mobility experiences. PMID- 11853062 TI - Modernization and religious intermarriage in the Republic of Ireland. AB - There are strong theoretical reasons for hypothesizing that those sections of the population who are most exposed to modernization processes are more likely to marry outside their own religious group. We examine this for Catholic-Protestant intermarriage in the Republic of Ireland. We use a multivariate analysis on survey data to test the hypotheses that urban dwellers, persons of non-farming parents, persons with higher levels of education and the young are more likely to be religiously intermarried. While the odds of being in an intermarriage are greater for urban dwellers, this is in large part due to their being non-farmers. The farming effect is not necessarily related to religiosity. The odds of being in an intermarriage do not increase significantly for persons with third level education and this can be explained in terms of the marriage market for minority groups. It is shown that the historical context and the minority position of religious group should be taken into account in explanations which relate modernization to the pattern of religious intermarriage. PMID- 11853063 TI - Critical realism and the dialectic. AB - A prominent strand within both sociological and social theory has been concerned to develop a 'systems approach' with which to explore social life. One of the most original contributions to a systems approach has arisen within critical realism. In particular critical realism demonstrates that it is possible to abstract the causal powers of different objects of analysis to examine their interaction within concrete and contingent 'open systems'. The recent dialectical turn of critical realism develops this systems approach in a much more rigorous manner. However, in this paper I argue that the (dialectical) critical realist mode of abstraction ultimately fails to embed concepts and categories internally within the specific ideological and historical forms of social relations. Or rather, critical realists do not seek to develop concepts that reflect the self movement of a historical and contradictory essence. This self-movement is what I prefer to call a 'system'. Consequently critical realists are led to separate method from system in theory construction and such a separation leads to a problematic dualist mode of theorizing. I make these observations from a Hegelian Marxist position. PMID- 11853064 TI - Values-intuitive rational action: the dynamic relationship of instrumental rationality and values insights as a form of social action. AB - This paper is concerned with the implications for conceptualizing social action which arise from a consideration of whether human beings are capable of knowing ultimate (universal, unconditionally valid) values. This issue is framed within the view that the validity of our understanding of social action is inextricably linked with the validity of our conception of humankind: the scope and variety of social action has potentialities and limitations that are inscribed by the nature of human beings qua human beings. The paper suggests an enlargement, through the addition of a proposed conceptual tool, of the framework that comprises Weber's typology of social action. It argues that the common human properties of the person in whom social action is embedded should not arbitrarily exclude questions of the genesis of values. An analytical argument is put forward through an examination of the extent to which a faculty for values insights is implicated in Weber's concept of charisma and ethical analysis of political leadership. The notion of values-intuitive rational action is then outlined and discussed. The analytical argument is supported by theorizing from developmental psychology and examples of such action are given. Methodological difficulties in investigating the latter and the interrelationship of such action with institutional and social contexts are briefly discussed. PMID- 11853065 TI - Abstract and engaged critique in sociology: on football hooliganism. AB - This note is a response to a critical review of some of my own work published recently in the British Journal of Sociology but it does not reply to the specific criticisms made in that article. Rather it addresses the question of what constitutes appropriate critique in sociology by developing a distinction between abstract and engaged criticism. PMID- 11853066 TI - Vascular calcifications as a footprint of increased calcium load and chronic inflammation in uremic patients: a need for a neutral calcium balance during hemodialysis? AB - Cardiovascular complications caused by an accelerated atherosclerotic disease represent the largest single cause of mortality in chronic renal failure patients. The rapidly developing atherosclerosis of the uremic syndrome appears to be caused by a synergism of different mechanisms, such as malnutrition, oxidative stress and genetic factors. Recent studies provide evidence that chronic inflammation plays an important role in the pathogenesis of cardiovascular diseases. Hyperphosphatemia and an increased calcium-phosphate ion product have also been associated with an increased risk of death. Cardiovascular calcifications secondary to increases in phosphate and calcium load in dialysis patients might exert an important contribution to the excess cardiovascular mortality and morbidity in dialysis patients. Elevated serum levels of plasma C reactive protein (CRP) are associated with the extent and severity of the atherosclerotic processes as well as with an increased risk of experiencing myocardial infarction and sudden cardiac death in apparently healthy subjects. In patients affected by pre-dialytic renal failure increased levels of CRP and IL-6 were recorded in 25% of our population; CRP and IL-6 were inversely related with renal function. These data suggest the activation--even in the predialytic phase of renal failure--of mechanisms known to contribute to the enhanced cardiovascular morbidity and mortality of the uremic syndrome. In recent years we have investigated the hypothesis that the chronic inflammatory state of the uremic patient could be at least in part due to the dialytic technique. We have shown that the increase of CRP in stable dialysis patients may be due to the stimulation of monocyte/macrophage by backfiltration of dialysate contaminants. During conventional dialysis, a positive calcium balance and a concomitant inflammatory state may act as cofactors in the development of cardiovascular calcifications. We suggest that this hypothesis should be verified by clinical studies. A reevaluation of the ideal calcium levels in the dialysate is warranted: a neutral intradialytic calcium balance is probably more appropriate, although not easily attainable. PMID- 11853067 TI - Cytokine removal with a large pore cellulose triacetate filter: an ex vivo study. AB - OBJECTIVE: To test the hypothesis that hemofiltration using a new large pore cellulose triacetate hemofilter can achieve effective ultrafiltration of cytokines. DESIGN: Ex-vivo study. SETTING: Laboratory of Intensive Care Unit in tertiary hospital. SUBJECTS: Six healthy volunteers. INTERVENTIONS: Blood from 6 volunteers was incubated for 4 hours with 1 mg of endotoxin and then circulated through a closed hemofiltration circuit with a large pore cellulose triacetate hemofilter (nominal cut-off point: 60 kilodaltons). Hemofiltration was conducted at 1 L/h or 6 L/h of ultrafiltrate (UF) flow at the start of extra-corporeal circulation, and after 2 and 4 hours. Samples were taken from the arterial, venous and UF sampling ports. MEASUREMENTS AND MAIN RESULTS: IL-Ibeta, IL-6, IL 8, IL-10, TNFalpha, and albumin were measured. Sieving coefficients (SC) above 0.6 were achieved for IL-Ibeta and IL-6 and SCs above 0.3 were achieved for IL-8 and TNF-alpha at 1 L/h. Sieving coefficients of all cytokines (except IL-10, p=0.22) were reduced when the ultrafiltration rate was increased from IL/h to 6 L/h (p<0.01), but the increase in ultrafiltration rate resulted in an overall increase in the clearance of all cytokines (p<0.001). The highest SC for albumin was 0.07 at 4 hours at 1 L/h, and fell to 0.01 at 6 L/h. The SCs for IL-8 fell at 4 hours (p<0.01), but the SCs for other cytokines did not change. No adsorption of cytokines and albumin was observed. CONCLUSION: High volume hemofiltration (HVHF) using a new large pore cellulose triacetate filter achieved cytokine clearances greater than those reported with currently available hemo filters. PMID- 11853068 TI - Peripheral vascular disease and ESRD: what is the most appropriate intervention? PMID- 11853069 TI - Clearance of quinupristin-dalfopristin (Synercid) and their main metabolites during continuous veno-venous hemofiltration (CVVH) with or without dialysis. AB - BACKGROUND: Quinupristin-dalfopristin (Q/D) is often utilized in critically ill patients, some of whom require CVVH. This study was undertaken to determine the clearance of O/D and their main active metabolites (RPR 100391, RP 69012, RP 12536) via CVVH in the swine model. METHODS: Q/D 7.5 mg/kg was intravenously administered over 0.5 h to 12 swine after induction of acute renal failure by ligation of the renal arteries. At 0.5 h post injection, the CVVH procedure was initiated and continued for 8 hours at the following pump rates: (1)100 mL/min, (2)180 rnL/min, and (3)100 mL/min with dialysis (flow rate: 1 L/h). Blood and ultrafiltrate samples were collected at 1 h intervals and assessed by a validated HPLC method. RESULTS: Plasma analysis suggests rapid metabolism to the main active metabolites which are appreciably cleared as demonstrated by high clearance and sieving coefficient estimates. Mean clearance estimates for RP 69012, RP 100391, and RP 12536 are 729, 777, and 578 mL/h in the 100 mL/min CVVH group, 772, 785, 685 mL/min in the 180 mL/min CVVH group, and 753, 791, 616 mL/min in the 100 mL/min CVVH group with 1 L/h dialysis, respectively. CONCLUSION: These data reveal that Q/D is rapidly metabolized and the metabolites are cleared to a large extent via CVVH. Due to the considerable contribution of the metabolites to overall in vivo activities, additional studies are required to fully quantify their removal before final dosage modifications for patients undergoing CVVH can be recommended. PMID- 11853070 TI - Randomized study of temporary hemodialysis catheters. AB - Temporary catheters are still essential for acute dialysis access but their performance has not been compared in randomized trials. We conducted a randomized trial of our standard catheter and a newly designed catheter. The primary outcome was maximized blood flow over the entire use of the catheter. Seventy-six patients provided maximum blood flows. The new catheter provided an average blood flow of 349 ml/min and the standard catheter provided flows of 320 ml/mm (p=0.09). Lumen reversal occurred in 56.8% of dialysis sessions with the standard catheter compared to 27.4% with the new catheter (p<0.001). Left-sided internal jugular catheters and catheters inserted in females provided 103 ml/min and 36 ml/min less blood flow, respectively, compared to right sided catheters and catheters placed in males. Approximately 20% of catheters were removed for poor blood flow. Left sided catheters and catheters in females were more likely to need removal for malfunction but catheter design did not influence removal rates. PMID- 11853071 TI - Extracorporeal pressure monitoring and the detection of vascular access stenosis. AB - Prospective monitoring of static venous pressure is an established tool to detect outflow stenoses in a vascular access. However, with this method it is not possible to identify vascular stenoses which are localized between the arterial and venous dialysis needle. We describe a new approach based on both static arterial and venous extracorporeal pressures. Pressure data of 9 dialysis patients with normal vascular access function and 9 patients with stenotic access were analyzed. Extracorporeal pressure was found to depend on the position of the heart relative to the extracorporeal blood circuit. All patients with venous outflow stenoses had an elevated ratio of arterial and venous intra-access pressure to mean arterial pressure. In case of access stenosis between arterial and venous needle the ratio of venous pressure to mean arterial pressure was normal, and only the arterial pressure ratio was elevated. We conclude that combined arterial and venous intraaccess pressure measurement normalized by mean blood pressure detects venous stenosis as well as stenosis between the arterial and venous dialysis needle. To minimize the rate of access thrombosis both arterial and venous intra-access pressure should be monitored. PMID- 11853072 TI - Development of a hybrid artificial liver using polyurethane foam/hepatocyte spheroid culture in a preclinical pig experiment. AB - We describe a preclinical study of our original hybrid artificial liver support system (HALSS) for a clinical trial. We designed a HALSS comprising a multi capillary polyurethane foam packed-bed module (MC-PUF module) containing a total 200 g (2 x 10(10) cells) porcine hepatocytes, and an extracorporeal circulation device. Almost all porcine hepatocytes in the MC-PUF module formed many spherical multicellular aggregates (spheroids). This extracorporeal circulation device was improved to promote solute exchange between a living body and a MC-PUF module by including a plasma bypass line in the circulation loop. The efficacy of the HALSS was evaluated using a 25-kg pig with warm ischemic liver failure by portocaval shunt and ligation of hepatic artery (HALSS group, n=3). As a control experiment, the same system without hepatocytes in the module was used with the same kind of liver failure pig (Control group, n=3). The blood ammonia in the control group was 143 N-microg/dl at the start of circulation, and rapidly increased to 351 N microg/dl at 2 hours and to 704 N-microg/dl at 6 hours. But the blood ammonia in the HALSS group was completely suppressed, and remained less than the hepatic coma level (over 200 N-microg/dl) during the circulation time. The blood glucose in the control group gradually decreased, and became less than 40 mg/dl within 6 hours of circulation. But the blood glucose in the HALSS group was maintained well, and remained the normal glucose level (50 - 105 mg/dl) for more than 20 hours of circulation. Improvement in blood creatinine and lactate, and the stabilization of vital signs and urinary excretion, were observed in the HALSS group. The survival time of the pigs in the HALSS group was 19.3 hours compared with 8.9 hours in the control group. In conclusion, our HALSS was effective to stabilize the general conditions of the body in addition to supporting various liver functions. These results suggest that our HALSS has a strong possibility to be used in treating liver failure patients. We have applied for approval of the clinical trial of our HALSS to our institutional ethics committee. PMID- 11853073 TI - Autologous cardiomyocyte transplantation in an ovine myocardial infarction model. AB - BACKGROUND: To evaluate the possibility of autologous cardiomyocyte transplantation, we transplanted cultured autologous cells into an infarct region developed by coronary artery ligation in an ovine model. MATERIALS AND METHODS: A chronic heart failure model with a considerable portion of myocardial infarction was created in sheep using sequential ligation of the homonymous artery and its diagonal branch. Autologous cardiomyocytes were cultured and isolated from the right ventricular infundibulum. After a predetermined interval (one animal for two months and the other for three months), the two animals were reanesthetized and a suspension of cultured autologous vetricular cells in 0.3 ml of culture medium (1.2 x 10(7) cells) was injected into the center of three out of the four sites in the infarcted area using a tuberculin syringe. The same amount of culture medium was injected with an identical procedure into the center of the remaining site, as control. The animals were kept alive for a further month, and then sacrificed for postmortem heart examinations. Light microscopic analysis and immunohistochemical study for myoglobin were done. RESULTS AND CONCLUSIONS: On postmortem gross examination, well-demarcated thin-walled anteroseptal infarcts with chamber enlargement were clearly seen in specimens from the two animals. Microscopic analysis showed homogenous fibrosis throughout the infarcted areas. In both animals, one of the three sites of cardiomycyte injection showed an islet of cardiomyocytes in the mid-myocardium, while none were observed in the control site of either animal. A layer of cardiomyocytes was observed in subendocardial regions, as it was in the control areas. In conclusion, cardiomyocyte transplantation into the infarct regions developed by coronary artery ligation in an ovine model was achieved with only limited success. An understanding of why only 33% of cardiomyocyte-injection sites demonstrated viable cardiomyocytes, in the form of tiny cell islets, remains to be elucidated. PMID- 11853074 TI - Magnification of the pore size in biodegradable collagen sponges. AB - In tissue engineering cells are often combined with a carrying structure with collagen being a suitable material to form a 3D-scaffold. A process to manufacture collagen sponges with an adjustable and homogeneous structure has been developed at the Helmholtz-Institute. Using this process, collagen suspensions are frozen directionally and subsequently vacuum-dried. One clinical application in which these scaffolds can be used is soft tissue reconstruction. Various soft tissue defects require an adequate replacement, e.g. in the case of severe burn wounds, or after tumour resections. Collagen (type I) sponges, which are cultured with preadipocytes, may be used to regenerate such defects. In this case, pore sizes of approximately 100 microm are desired to allow a complete differentiation of preadipocytes into adipocytes. Based on known technology to manufacture collagen sponges with an adjustable and homogeneous pore structure, research on the increase of pore size beyond the previous limit of 40 microm was necessary in order to enable soft tissue replacement. A scaffold with an average pore size of 100 microm was obtained. PMID- 11853075 TI - Epidemiology of hepatitis delta virus (HDV) infection in the dialysis population. PMID- 11853077 TI - Evaluation of quadriceps strength and voluntary activation after unicompartmental arthroplasty for medial osteoarthritis of the knee. AB - INTRODUCTION: In early and moderate stages of osteoarthritis (OA) of the knee, arthrogenous muscle inhibition (AMI) is an important factor for the initiation and the progression of the disease. Although AMI has been shown to be reduced after physiotherapeutical exercises resulting in significant improvements in disability, implantation of unicondylar knee arthroplasties is much provided in these stages of OA. Therefore, in the present study we investigate changes in quadriceps muscle after implantation of such prostheses as compared to physiotherapeutical treatment, alone. METHODS: In eighteen patients with bilateral moderate knee OA, who were treated with unicondylar knee arthroplasty we investigated voluntary activation (VA) and maximum voluntary contraction (MVC) of the quadriceps femoris muscle. There were 7 males and 11 females, the mean age at time of operation was 67 years (range 58-76 years). Measurements on both sides were performed preoperatively and 18 months postoperatively using the twitch interpolation technique. RESULTS: Follow-up assessment revealed a significant VA and MVC increase in both the surgically treated knees and in the contralateral knees treated by physiotherapy alone. However, VA and MVC improvements were significantly higher in the operated on knees than in those treated by physiotherapy alone. DISCUSSION: Both physiotherapeutical exercise and unicondylar knee replacements lead to an improvement of quadriceps motor function in knee OA. The greater improvement in knees with both knee replacement and physiotherapy might be related to the intraoperative removal of arthritic tissue in these knees. PMID- 11853076 TI - The knee adduction moment during gait in subjects with knee osteoarthritis is more closely correlated with static alignment than radiographic disease severity, toe out angle and pain. AB - This study tested whether the peak external knee adduction moments during walking in subjects with knee osteoarthritis (OA) were correlated with the mechanical axis of the leg, radiographic measures of OA severity, toe out angle or clinical assessments of pain, stiffness or function. Gait analysis was performed on 62 subjects with knee OA and 49 asymptomatic control subjects (normal subjects). The subjects with OA walked with a greater than normal peak adduction moment during early stance (p = 0.027). In the OA group, the mechanical axis was the best single predictor of the peak adduction moment during both early and late stance (R = 0.74, p < 0.001). The radiographic measures of OA severity in the medial compartment were also predictive of both peak adduction moments (R = 0.43 to 0.48, p < 0.001) along with the sum of the WOMAC subscales (R = -0.33 to -0.31, p < 0.017). The toe out angle was predictive of the peak adduction moment only during late stance (R = -0.45, p < 0.001). Once mechanical axis was accounted for, other factors only increased the ability to predict the peak knee adduction moments by 10 18%. While the mechanical axis was indicative of the peak adduction moments, it only accounted for about 50% of its variation, emphasizing the need for a dynamic evaluation of the knee joint loading environment. Understanding which clinical measures of OA are most closely associated with the dynamic knee joint loads may ultimately result in a better understanding of the disease process and the development of therapeutic interventions. PMID- 11853078 TI - Active range of motion of the head and cervical spine: a three-dimensional investigation in healthy young adults. AB - PURPOSE: To define reference values for head-cervical range of motion (ROM) in healthy young adults, to assess the effect of sex, and to quantify the separate contribution of other body districts. METHODS: Thirty women and 30 men performed maximal head and cervical spine flexion-extension, lateral bending, and axial rotation. Movements were detected using a digital optoelectronic instrument. Maximum head-cervical spine and thoracic motions were separated. RESULTS: Flexion and extension were performed mainly in the sagittal plane. The movement was larger in women (136 degrees) than in men (130 degrees). During flexion, both sexes moved the head-neck and the thorax in the same direction. During extension, men moved only the head-cervical spine, while women moved the two analyzed districts in the opposite directions. Lateral bending was nearly symmetric, associated with head-cervical rotation and extension, and larger in women (91 degrees) than in men (77 degrees). Adjunctive thoracic motion was limited in the sagittal and frontal planes, but larger in the horizontal plane (opposite motions of about 20 degrees). Head-neck rotation was symmetric, and associated with concomitant movements in both the sagittal and frontal planes. It was larger in women (162 degrees) than in men (155 degrees), and performed with limited adjunctive thoracic motions. CONCLUSIONS: The present values can be used as a first group of normative data for head-cervical ROM in young men and women. PMID- 11853079 TI - Significance of lesion size and location in the prediction of collapse of osteonecrosis of the femoral head: a new three-dimensional quantification using magnetic resonance imaging. AB - Size and location of a necrotic lesion are considered important factors predicting collapse of the femoral head in the early stages of osteonecrosis. However, few analytical studies have performed a three-dimensional quantification of lesions to clarify how these two morphological factors are related to the occurrence of collapse. We evaluated the relevance of lesion size and location for prediction of collapse quantitatively using new three-dimensional indexes. Magnetic resonance (MR) imaging was performed in 65 hips in a consecutive series of 47 patients with osteonecrosis without radiological evidence of collapse. Lesion volume as well as latitude and longitude of the center of gravity of the lesion within the femoral head were calculated. Thirty-three hips developed radiological collapse, while in the remaining 32 hips collapse did not occur over 2 years. Multiple logistic regression analysis showed a significant relationship between lesion volume and radiological collapse. In 35 hips in which the lesion volume was less than 30% of the femoral head, only 9 collapsed. In comparison with non-collapsed hips, collapsed hips had a significantly higher combined value for latitude and longitude of the lesion, corresponding to the anterosuperior portion of the femoral head. Quantitative analysis of lesion morphology demonstrated that lesion volume is strongly correlated with risk of collapse, and that lesion location is an important prognostic indicator of collapse in small necrotic lesions. PMID- 11853080 TI - Tissue response during monofocal and bifocal leg lengthening in patients. AB - The purpose of this investigation was to compare the tissue response during mono- and bifocal limb lengthening. The study includes four patients undergoing leg lengthening. All patients started out bifocally with a total diurnal distraction of 1.75 mm, but proceeded monofocally with a rate of 1 mm a day when the distal distraction was terminated due to contractures or pain. The tissue response was monitored by registration of axial force in the distraction rods. The force increased linearly during bifocal lengthening, but culminated or decreased in the period of monofocal lengthening. Average tissue stiffness, defined as the immediate force increase due to each 0.25 mm distraction increment, was significantly higher in the bifocal lengthening phase. The force decay between each distraction was significantly lower during bifocal lengthening, thus indicating decreased tissue accommodation. Details in the force registrations indicated that the soft tissue, not the regenerate, was the main contributor to the tensile force. Conclusively, the tissues at the two osteotomy sites do not lengthen independently. Bifocal lengthening exposes the entire soft tissue to large loads, resulting in increased tissue stiffness and reduced ability to adapt to the increased length. Accordingly, bifocal leg lengthening requires special attention to soft tissue adaptation. PMID- 11853081 TI - Osteogenic protein-1 induced bone formation in an infected segmental defect in the rat femur. AB - The goal of this study was to use a segmental defect model in the rat femur to determine if osteogenic protein-1 (OP-1) is capable of inducing bone formation in the presence of bacterial contamination. A 6 mm segmental defect was surgically created and stabilized with a polyacetyl plate and Kirschner wires in one femur in each of 126 Sprague-Dawley rats. The animals were divided into eight groups in which the defect was either left untreated, or subjected to various combinations of OP-1 (11 or 50 microg), lyophilized bovine type I collagen (carrier for the OP 1), and 10(5) colony-forming units of Staphylococcus aureus. The animals were euthanized at either 2, 4, or 9 weeks. Quantitative radiographic and histologic analyses were performed on the harvested tissue. The initial contamination progressed to infection in all animals receiving bacteria, as determined by qualitative bacteriology. There was very little, if any, bone formation in the untreated defects, and in the contaminated defects with or without collagen carrier. Bone formation was significantly greater in contaminated defects with either dose of OP-1, compared with contaminated defects without OP-1. The 50 microg dose of OP-1 induced significantly more bone formation than the 11 microg dose, both with and without bacteria. This investigation has demonstrated that OP 1 maintains its osteoinductive capability in a contaminated segmental defect. OP 1 may potentially be used in the clinical management of contaminated fractures. PMID- 11853082 TI - Measurement of material elastic constants of trabecular bone: a micromechanical analytic study using a 1 GHz acoustic microscope. AB - The propagation speed (C) of surface acoustic waves (SAW), e.g. Rayleigh (R waves) and longitudinal lateral waves (L-waves), the latter being the surface manifestation of the longitudinal waves, strongly reflect mechanical properties of materials. In view of an increasing interest in ultrasonic methodology in the field of bone biomechanics, we tested the hypothesis that both R- and L-waves can be excited in trabecular bone using an acoustic microscope at 1 GHz and that their speeds (C(R) and C(L)) can be extracted from V(z)-curves, i.e. plots of lens output voltage as a function of the lens focal point position with respect to the specimen surface. In accordance with V(z)-curves theoretically synthesized on the basis of incident field theory, experimental curves for canine femoral trabecular bone showed evidence of both R- and L-waves in almost all regions of recording. The measured CR ranged between 1.93 and 2.07 km/s (mean +/- SD.: 2.00=0.06 km/s) and the C(L) ranged between 2.33 and 4.33 km/s (3.37+/-0.61 km/s). Knowledge of both speeds allowed computation of a number of material constants by means of simple theory of elasticity and assumptions of the material density. We found values of Poisson ratio (v) ranging from 0.14 to 0.32 (0.23+/ 0.07). Young's modulus (E) from 15 to 22.8 GPa (19.9+/-2.5 GPa) and the shear modulus (G) from 7.6 to 8.9 GPa (8.4+/-0.5 GPa). Anisotropy in the trabecular bone material was clearly detected at the micrometer level. In conclusion, the V(z)-curve method was successfully used to determine the distribution of material elastic constants of trabecular bone with micrometer resolution. PMID- 11853083 TI - COX-1 and COX-2 expression in osteoid osteomas. AB - Osteoid osteoma is a benign bone forming neoplasm that is characterized by its small size (less than 2 cm), self-limited growth, and the tendency to cause extensive reactive changes in the adjacent tissue. The lesion classically presents with severe pain at night that is dramatically relieved by NSAIDs. The tumor has been shown to express very high levels of prostaglandins, particularly PGE2 and PGI2. The high local levels of these prostaglandins are presumed to be the cause of the intense pain seen in patients with this lesion. One generally accepted form of treatment is the prolonged use of NSAIDs. Since the cyclooxygenases are thought to be the source of these prostaglandins, and the central target of NSAIDs, we evaluated the expression of cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2) in osteoid osteoma tissues from patients following surgery. In the 12 specimens examined we found that the tumor osteoblasts had strong immunohistochemical staining for COX-2, while the staining in the surrounding host osteoblasts in the reactive bone was scant. Significant COX-1 staining was also detected in both tumor and host osteoblasts. For comparison we examined the COX expression in human fracture callus, fibrous dysplasia, osteoblastoma, osteofibrous dysplasia, and myositis ossificans. With the exception of fracture callus, very limited amounts of COX-2 could be detected in these tissues. Taken together, we conclude that the increased production of prostaglandins by osteoid osteomas implicates that COX-2 is one of the mediators of this condition. These findings suggest that the newly selective COX-2 inhibitors could be used to more safely treat osteoid osteomas. PMID- 11853084 TI - A biodegradable composite scaffold for cell transplantation. AB - Cell transplantation is rapidly becoming a therapeutic option to treat disease and injury. However, standard techniques for cell seeding on non-woven polymer meshes or within gels may not be suitable for immediate implantation or surgical manipulations of freshly isolated cells. Therefore, a biodegradable composite system was developed as a way to rapidly entrap cells within a support of predefined shape to potentially facilitate cell delivery into a target site (e.g. meniscal tears in the avascular zone). The composite construct consisted of freshly isolated cells, in this case pig chondrocytes, entrapped in a fibrin gel phase and dispersed throughout the void volume of a polyglycolic acid (PGA) non woven mesh. Composites were cultured for up to 4 weeks. In vitro degradation of fibrin gel was evaluated via gel-entrapped urokinase. At 28 days in culture, glycosaminoglycan (GAG) content per cell in the composite scaffolds was 2.6 times that of the PGA-only cell construct group and 88% that of native pig cartilage. Total collagen content per cell in the composite scaffolds was not significantly different from the PGA-only cell construct group (P > 0.02) and represented 40% of the value determined for native cartilage. Varying the concentration of entrapped urokinase could effect controlled degradation of fibrin gel. PMID- 11853085 TI - Androgen receptors in Dupuytren's contracture. AB - Palmar fascia tissue and cultured cells from patients with Dupuytren's contracture and from normal subjects were characterized and analyzed for androgen receptor expression. Androgen receptors have never been studied in Dupuytren's myofibroblasts and may have a role in its high male predominance. Surgical samples were collected from eight patients undergoing surgery for Dupuytren's contracture and from four patients with carpal tunnel syndrome, used as control tissue. Immunohistochemical analysis was performed on tissue samples and on cell cultures with anti-androgen receptor, anti-alpha-smooth muscle actin, anti fibronectin, and anti-type I and III collagen antibodies using the biotin avidin peroxidase method as revelatory system. Immunostaining for androgen receptors in tissue samples and cultured cells revealed nuclear reaction in many Dupuytren's myofibroblasts, but in few fibroblasts of the normal palmar fascia. In a double labeling study, androgen receptors were seen to co-localize with alpha-actin in both cell cultures and tissue samples. We present the first evidence that the palmar fascia is a target tissue for androgen action and that the expression of androgen receptors in Dupuytren's contracture is considerably higher than in the normal palmar fascia. Further studies will need to evaluate whether the androgen responsive state of the tissue is related to the high incidence of Dupuytren's contracture in the male sex. PMID- 11853086 TI - Integrin expression by human osteoblasts cultured on degradable polymeric materials applicable for tissue engineered bone. AB - The use of biodegradable polymers in the field of orthopaedic surgery has gained increased popularity, as surgical pins and screws, and as potential biological scaffolds for repairing cartilage and bone defects. One such group of polymers that has gained considerable attention are the polyesters, poly(lactide-co glycolide) (PLAGA) and polylactic acid (PLA), because of their minimal tissue inflammatory response, favorable biocompatibility and degradation characteristics. The objective of this study was to evaluate human osteoblastic cell adherence and growth on PLAGA and PLA scaffolds by examining integrin receptor (alpha2, alpha3, alpha4, alpha5, alpha6 and beta1) expression. Primary human osteoblastic cells isolated from trabecular bone adhered efficiently to both PLAGA and PLA, with the rate of adherence on PLAGA comparable to that of control tissue culture polystyrene (TCPS), and significantly higher than on PLA polymers at 3, 6 and 12 h. Human osteoblastic phenotypic expression, alkaline phosphatase (ALP) activity was positive on both degradable matrices, whereas osteocalcin levels were significantly higher on cells grown on PLAGA than on PLA composites. Interestingly, the integrin subunits, alpha2, alpha3, alpha4, alpha5, alpha6 and beta1 were all expressed at higher levels by osteoblasts cultured on PLAGA than those on PLA as analyzed by westerns blots and by flow cytometry. Among the integrins, alpha2, beta5 and beta1 showed the greatest difference in levels between the two surfaces. Thus, both PLA and PLAGA support osteoblastic adhesion and its accompanying engagement of integrin receptor and expression of osteocalcin and ALP. However PLAGA consistently appeared to be a better substrate for osteoblastic cells based on these parameters. This study is one of the first to investigate the ability of primary human osteoblastic cells isolated from trabecular bone to adhere to the biodegradable polymers PLAGA and PLA, and to examine the expression of their key adhesion receptors (integrins) on these substrates. PMID- 11853087 TI - In situ cell nucleus deformation in tendons under tensile load; a morphological analysis using confocal laser microscopy. AB - Cell and cell nucleus deformations have been implicated in the mechanotransduction of mechanical loads acting on tissues. While in situ cell nucleus deformation in response to increasing tissue strains has been examined in articular cartilage this phenomenon has not been investigated in tendons. To examine in situ cell nuclei deformation in tendons undergoing tensile strain rat tail tendons were harvested from adult Sprague-Dawley rats and stained with acridine orange to highlight the cell nuclei. The tendons were mounted on a custom-designed, low-load, tensile testing device affixed to the mechanical stage of a confocal laser microscope. Cells within the tendons were isolated for analysis. Images of individual cells were captured at 0% strain as well as sequentially at 2%, 4% and 6% grip-to-grip tendon strain. Digital images of the cell nuclei were then measured in the x (length) and y (height) axis and deformation expressed as a percentage of cell nuclei strain. In addition, centroid-to-centroid distances of adjacent cell nuclei within each image were measured and used to calculate local tissue strain. There was a weak (r2 = 0.34) but significant (p < 0.01) correlation between local tissue strain and cell nucleus strain in the x axis. The results of this study support the hypothesis that in situ cell nucleus deformation does occur during tensile loading of tendons. This deformation may play a significant role in the mechanical signal transduction pathway of this tissue. PMID- 11853088 TI - Stretch and interleukin-1beta induce matrix metalloproteinases in rabbit tendon cells in vitro. AB - Little is known about the factors that initiate and propagate tendon overuse injuries, but chronic inflammation and matrix destruction have been implicated. The purpose of this study was to evaluate the production of cyclooxygenase II (COX-2) and matrix metalloproteinases (MMPs) by tendon cells exposed to cyclic strain and inflammatory cytokines in vitro. Rabbit Achilles tendon cells were subjected to a stretching protocol with 5% elongation at 0.33 Hz for 6 h, or treated with 1000 pM interleukin-1beta (IL-1beta), or exposed to IL-1beta and stretching together. Gene expression was evaluated by RT-PCR and production of stromelysin was quantified with an ELISA. IL-1beta induced the expression of the collagenase-1 and stromelysin-1 genes. Production of stromelysin proenzyme by cells stimulated with IL-1beta was 17 times higher than production by control cells. Cells exposed to IL-1beta and stretching produced 20 times more stromelysin than control cells. Cells subjected to stretching alone did not produce more stromelysin than control cells. The synergistic effect of IL-1beta and stretching was observed at doses of IL-1beta ranging from 10 to 1000 pM. These data suggest that mechanical load and inflammatory cytokines can initiate a matrix destructive pathway in tendon that is more pronounced than with mechanical loading or inflammation alone. PMID- 11853089 TI - Low frequency EMF regulates chondrocyte differentiation and expression of matrix proteins. AB - This study describes the enhancement of chondrogenic differentiation in endochondral ossification by extremely low frequency pulsed electric/magnetic fields (EMFs). The demineralized bone matrix (DBM)-induced endochondral ossification model was used to examine the effects of EMF stimulation. [35S] Sulfate and [3H]-thymidine incorporation and glycosaminoglycan (GAG) content were determined by standard methods. Proteoglycan (PG) and GAG molecular size and composition were determined by gel chromatography and sequential enzyme digestion. Immunohistochemical and Western blot analysis of PGs were done with antibodies 2B6, 3B3, 2D3 and 5D4. Northern analysis of total RNA extracts was performed for aggrecan, and type II collagen. All data was compared for significance by Student's t- or analysis of variance (ANOVA)-tests. The EMF field accelerated chondrogenesis as evidenced by an increase in: (1) 35SO4 incorporation and GAG content, (2) the number of chondrocytes at day 8 of development, (3) the volumetric density of cartilage and (4) the extent of immunostaining for 3B3 and 5D4. No differences in DNA content or [3H]-thymidine incorporation were observed between control and stimulated ossicles, suggesting the absence of enhanced cell proliferation or recruitment as a mechanism for the acceleration. PG and GAG molecular sizes and GAG chemical composition were similar in stimulated and control ossicles, indicating that stimulation resulted in an accelerated synthesis of normal cartilage molecules. The increased expression of PG and type II collagen mRNA as well as a greater immunoreactivity of 3B3 and 5D4 suggest an increase in the rate of differentiation of chondrocytes and enhanced phenotypic maturation. PMID- 11853090 TI - Calcium signaling is required for ultrasound-stimulated aggrecan synthesis by rat chondrocytes. AB - Low-intensity ultrasound accelerates fracture healing in humans. In rat femur fracture models, ultrasound advanced healing is associated with increased proteoglycan expression. Here we report that ultrasound stimulation of primary rat chondrocytes elevated the intracellular concentration of calcium [Ca2+]i. The [Ca2+]i increase was rapid and transient at lower pressures (175-320 kPa), but rapid and sustained at higher ultrasound exposures (350-500 kPa). Chelating internal [Ca2+]i with 1,2-bis(2-aminophenoxy) ethane-N-N-N',N'-tetraacetic acid (BAPTA-AM), stopping the Ca2+/ATP-ase induced mitochondrial release of [Ca2+]i with Thapsigargin, or removing [Ca2+]i from the medium with EGTA inhibited the stimulatory effects of ultrasound on proteoglycan synthesis. These results imply that ultrasound-stimulated synthesis of cell matrix proteoglycan, associated with accelerated fracture healing, is mediated by intracellular calcium signaling. PMID- 11853091 TI - Temporal expression patterns of BMP receptors and collagen II (B) during periosteal chondrogenesis. AB - Articular cartilage has a limited ability to repair itself. Periosteal grafts have chondrogenic potential and are used clinically to repair defects in articular cartilage. An organ culture model system for in vitro rabbit periosteal chondrogenesis has been established to study the molecular events of periosteal chondrogenesis in vitro. In this model, bone morphogenetic protein-2 (BMP2) mRNA expression was found to be upregulated in the first 12 h. BMPs usually transduce their signals through a receptor complex that includes type II and either type IA or type IB BMP receptors. Receptors IA and IB play distinct roles during limb development. We have examined the temporal expression patterns for the mRNAs of these receptors using our experimental model. The mRNA expression patterns of these three BMP receptors differed from one another in periosteal explants during chondrogenesis. When these explants were cultured under chondrogenic conditions (agarose suspension with TGF-beta1 added to the media for the first 2 days), the expression of BMPRII mRNA and that of BMPRIA mRNA varied only slightly and persisted over a long time. In contrast, the expression of BMPRIB mRNAwas upregulated within 12 h, peaked at day 5, and fell to a level that was barely detected beyond day 21. Moreover, the expression of BMPRIB mRNA preceded that of collagen type IIB mRNAs, a marker for matrix-depositing chondrocytes. These data support a role for coordinate expression of BMP2 and its receptors early during periosteal chondrogenesis. PMID- 11853092 TI - The involvement of beta1 integrin in the modulation by collagen of chondrocyte response to transforming growth factor-beta1. AB - The physiologic response of chondrocytes to maintenance of the matrix and response to injury likely involves signaling from multiple sources including soluble cytokines, mechanical stimulation, and signaling from the extracellular matrix. The signaling from the extracellular matrix may serve to effect cell differentiation and to modulate the response to cytokines. We have previously reported that type II collagen modulates the response of bovine articular chondrocytes to TGF-beta1. The molecular nature of the signaling mechanism has not been elucidated but presumably involves a similar mechanism by which the cell attaches to the surrounding matrix. An alginate bead culture system is utilized to which exogenous type II collagen is added. The inclusion of type II collagen results in an alteration of integrin expression with a down regulation of alpha2. The response of the chondrocyte to TGF-beta1 can be modulated by the inclusion of exogenous type II collagen. The modulation of DNA and proteoglycan synthesis was blocked by the treatment of anti-beta1 integrin antibody (4B4) or by cyclic RGD containing peptides. These events occur at concentrations that block cell adhesion to type II collagen. Linear RGD containing peptides and anti-anchorin antibodies had no effect on the modulation by type II collagen. These results suggest that type II collagen binding by chondrocytes at least in part occurs through the beta1 integrin. This binding results in modulation of the cell response to TGF-beta1. This modulation may serve to provide physiologic specificity to the cytokine-signaling cascade. An understanding of the regulatory milieu of the chondrocyte may permit the stimulation of an intrinsic repair of articular cartilage in the future. A near term application of this understanding can be made to tissue engineering attempts at articular cartilage repair. PMID- 11853094 TI - Altered swelling behavior of femoral cartilage following joint immobilization in a canine model. AB - Periods of reduced joint loading have been shown to induce changes in the biochemical composition. metabolism and mechanics of articular cartilage. In this study, changes in cartilage swelling behavior were studied following a 4-week period of joint immobilization, using a recently developed osmotic loading technique [J. Biomech, 32 (1999) 401-408]. The magnitude and distribution of swelling strains were measured in cartilage-bone samples equilibrated in physiological and hypotonic saline, relative to a hypertonic reference NaCl solution. Physicochemical parameters (glycosaminoglycan fixed charge density and water volume fraction) were determined in site-matched cartilage samples. The experimental data for swelling strains, fixed charge density and water volume fraction were used with a triphasic mechano-chemical theory [J. Biomech. Eng. 113 (1991) 245-258] to determine the effect of joint immobilization on the tensile modulus of the cartilage solid matrix. Four weeks of immobilization resulted in a significant increase in the magnitude of swelling-induced strains, and a significant decrease in fixed charge density in cartilage, as compared with the contralateral controls. Joint immobilization also resulted in decreases in values for the modulus of cartilage, as compared with the contralateral controls. Our results suggest that 4 weeks of joint immobilization had a significant effect on cartilage mechanical function that may be linked to collagen changes in the cartilage extracellular matrix. PMID- 11853093 TI - Tissue transglutaminase localization and activity regulation in the extracellular matrix of articular cartilage. AB - Tissue transglutaminase (tTG) catalyzes a Ca2+-dependent transglutaminase (TGase) activity which cross-links proteins and stabilizes many tissues [C.S. Greenberg et al. FASEB J. 5 (1991) 3071]. Because cartilage is subjected to great stress in vivo, an enzyme that strengthens and stabilizes tissue could play an integral role in maintaining cartilage integrity. The purpose of this study was to determine if active tTG is present in the extracellular matrix (ECM) of adult human osteoarthritic articular cartilage. Using a TGase activity assay along with immunolabeling for tTG of cartilage sections, TGase activity and tTG immunoreactivity were localized in the ECM in cartilage sections, predominantly in the superficial layer. Previous in vitro studies have demonstrated that the Mg GTP complex inhibits the TGase activity of tTG [T.S. Lai et al. J. Biol. Chem. 273 (1998) 1776]. To investigate the in situ regulation of the TGase activity of tTG, a TGase activity assay was done with a dose response of GTP, measuring incorporation of fluorescein cadaverine. TGase activity was inhibited by GTP in a similar manner as in vitro. These results not only confirm tTG presence in the ECM. but also indicate tTG as the major TGase activity of the ECM. Secondly, the study provides a possible mechanism by which extracellular tTG is regulated in vivo. PMID- 11853095 TI - Differential apoptotic response of J774 macrophages to alumina and ultra-high molecular-weight polyethylene particles. AB - We recently identified apoptosis in in vitro wear particle-stimulated macrophages. The recent explosion of interest in apoptosis lies in the fact that it is under positive and negative regulation through evolutionary conserved biochemical pathways. It may also be possible to modulate macrophage apoptosis in the treatment of periprosthetic osteolysis. The purpose of this study was to compare the macrophage response to identically sized particles of alumina ceramic (Al2O3) and ultra-high-molecular-weight polyethylene (UHMWPE) in terms of TNF alpha release and induction of apoptosis. J774 mouse macrophages were incubated for 0-24 h in the presence of Al2O3 and UHMWPE particles. TNF-alpha release was measured by ELISA; Poly(ADP-ribose)polymerase (PARP) and caspase-3 expression was analyzed by Western blot; DNA fragmentation (DNA laddering) was visualized on agarose gel containing ethidium bromide. Al2O3 particles induced TNF-alpha release after 4 h incubation with concentrations reaching 483 and 800 pg/ml after 24 h with 125 and 250 particles/macrophage, respectively (control = 161 pg/ml) (P < 0.05 vs. control). The same concentrations of UHMWPE particles induced a much larger and significant TNF-alpha release after only 1 h incubation, increasing up to 6250 pg/ml after 24 h (P < 0.05 vs. control). Western blot analysis demonstrated that the active caspase-3 fragment (17 kDa) and the proteolytic PARP fragment (85 kDa) were expressed after 2 h incubation with 125 and 250 Al2O3 particles/macrophage. The active caspase-3 and the PARP fragment had lower expression and appeared after a longer incubation time (8 h) with 125 and 250 UHMWPE particles/macrophage. Finally, DNA fragmentation (DNA laddering) was observed after 16 h with 125 and 250 particles of Al2O3 per macrophage whereas no laddering was induced by UHMWPE particles even after 24 h incubation. This study shows that although both Al2O3 and UHMWPE particles induce TNF-alpha release, this stimulation was much greater (8-10 times higher) with UHMWPE than Al2O3 (P < 0.05 vs. control). As well, the induction of apoptosis, as measured by activation of caspase-3, PARP cleavage and DNA laddering, is different for these two particles, being faster and more important with Al2O3 than UHMWPE. We hypothesize that the ability of Al2O3 to induce macrophage apoptosis may explain the lower TNF-alpha release observed with these particles and explain the differences seen in osteolysis patterns of ceramic-ceramic (CC) vs. metal-polyethylene (Mpe) articulations. In conclusion, apoptosis may be a major internal mechanism to decrease macrophage activity and may be a desired therapeutic endpoint. The identification of an apoptosis-related pathway in the macrophage response to ceramic particles provides crucial data for a rational approach in the treatment and/or prevention of periprosthetic osteolysis. PMID- 11853097 TI - Evolutionary psychiatry. Adaptationist and nonadaptationist conceptualizations. AB - Darwin's theory of evolution, and in particular one of its mechanisms, natural selection, is being used as the explanatory cornerstone of many unsolved problems in human biology and human affairs. Psychiatry is an example of that. Darwinian psychiatry's main proponents endorse the adaptationist program to carry out their project to implement an evolutionary psychiatry. The adaptationist program is an attempt to view all evolutionary novelties as adaptations, i.e., classically, features that favour survival and/or reproduction. This position is definitely teleological, and anthropomorphism plays a central role in its construction. This paper takes issue with the adaptationist approach. We argue that organism environment interactions are bidirectional processes. Hence, as a result of the fact that "a surprisingly large amount of the environment, which affects natural selection on an animal is the more or less direct result of the animals own behavior" [Waddington, C.H., 1976. Evolution of the subhuman world. In: Jantsch, E., Waddington, C.H. (Eds.), Evolution of Consciousness. London, UK, pp. 11-23], a more appropriate term to describe these interactions appears to be construction rather than adaptation alone [Lewontin, R., 2000. The triple helix: gene organism and environment. Harvard Univ. Press]. We present factual anatomical, physiological and clinical data critical of the platonic Kraepelinean classification of mental diseases, and claim that this classification is contrary to modern ideas on the evolution of nervous systems. We argue against the view of mainstream evolutionary psychiatrists that mental diseases are adaptations. We do so on two accounts. One is methodological; authors in this position do not ask whether every disease has evolutionary causes, but assume this in order to explain all diseases in such terms. The other mistake is biological; it is their belief that adaptation is the driving force of evolution while in fact it is just an outcome of evolution. The current status of the controversy between cognitive versus emotional experiences as essentially independent is reviewed, and evidence is presented, that they cannot be considered platonic, categorically independent functions of CNSs. These data, taken together, plus arguments derived from the high degree of plasticity of nervous systems, lead us to suggest a different approach to classification of mental diseases. PMID- 11853096 TI - Synovial fluid biomarker levels predict articular cartilage damage following complete medial meniscectomy in the canine knee. AB - The purposes of this study were to document the histological changes present in the tibial plateaus 12 weeks after complete medial meniscectomy in dogs and to determine if synovial lavage fluid biomarker levels are predictive of the severity of joint damage. Twelve adult dogs underwent complete unilateral medial meniscectomy and synovial lavage fluid biomarker levels, including cartilage oligomeric matrix protein (COMP), keratan sulfate (5D4). 3B3(-), and 3B3(+), were measured serially at 4-week intervals. The dogs were euthanized 12 weeks after surgery and each medial and lateral tibial plateau from the meniscectomized and contralateral knees was graded histologically. Histological data were analyzed using principal components analysis, which resulted in 4 factors that explained 70% of the variation in the data. Factor 2 (weighted most heavily by subchondral bone thickness) and Factor 3 (representative of articular cartilage damage) were significantly affected by compartmental site (P < 0.01 for both). Both of these factors were highest in the medial tibial plateau of the meniscectomized knee, and Factor 3 was significantly higher in this site than in the medial tibial plateau of the contralateral knee (P < 0.01). Peak levels of all 4 synovial lavage fluid biomarkers occurred at 4 weeks post-meniscectomy and 4-week minus baseline levels of all biomarkers were significantly correlated with the Factor 3 scores. This study demonstrates that significant articular cartilage damage occurs relatively quickly following complete medial meniscectomy in dogs and establishes the content and criterion validity for these synovial fluid lavage biomarkers in canine meniscectomy as surrogate measures of articular cartilage damage. PMID- 11853099 TI - Effect of repetitive transcranial magnetic stimulation on forced swimming test. AB - Repetitive transcranial magnetic stimulation (rTMS) and electroconvulsive shock (ECS) have been shown to affect mood in health and disease. Evidence to date has demonstrated an antidepressant potential for rTMS and electroconvulsive therapy (ECT). The present experiment, aimed at comparing the effects of ECS and rTMS in rats, employed one test used for screening of antidepressant activity: the forced swimming test (FST). In this study, the authors investigated whether chronic rTMS influenced active behavior in the rat FST, similar to ECS. Male Wistar rats received rTMS treatment daily, for 10 days as is commonly used for ECT treatment. Control rats received sham treatment by placing the stimulation coil in a perpendicular position to the rat's head. Passing a current through earclip electrodes for 1 s induced ECS. The control animals were treated identically, but current was not applied. The FST was carried out 24 h after the last rTMS or ECS. The immobility time in the FST was not significantly affected by rTMS and ECS for 1 day. The immobility time in the FST was significantly shortened at rTMS and ECS for 10 days. Chronic treatment with rTMS, similar to chronic treatment with ECS, decreased the immobility time in the FST. These results indicate that chronic treatment with rTMS might have antidepressant effect similar to chronic treatment with ECS. PMID- 11853098 TI - Intracerebroventricular antisense to inositol monophosphatase-1 reduces enzyme activity but does not affect Li-sensitive behavior. AB - Inositol monophosphatase (IMPase) inhibition is a hypothesized mechanism of action of lithium (Li). To test this hypothesis, the authors used the approach of antisense administration. Three days of an intracerebroventricular (icv) administration of 5 microg/20 microl 3'-phosphorothioated IMPA-1 antisense oligonucleotide sequence resulted in 20% reduction of rat periventricular IMPase activity. Li potentiates pilocarpine-induced seizures, because inhibition of IMPase leads to reduction in brain inositol levels. However, antisense-induced reduction in IMPase activity was not followed by seizures induced by subconvulsive pilocarpine doses. PMID- 11853100 TI - Chronic stimulation of the cat vagus nerve: effect on sleep and behavior. AB - The effect of electrical vagus nerve stimulation (VNS) on sleep and behavior was analyzed in freely moving cats. Eight cats were prepared for 23-h sleep recordings. The left vagus nerve of four of them was stimulated during 1 min, five times at 1-h intervals, for 5 days. The VNS induces: ipsilateral myosis, blinking, licking, abdominal contractions, upward gaze, swallowing, and eventually yawning and compulsive eating, as well as an increase of ponto geniculate-occipital (PGO) wave density and of the number of stages and total amount of rapid eye movement (REM) sleep. Besides, there was a sudden transition from waking stage to REM sleep. The present results suggest that VNS modifies sleep in the cat. This effect could be explained by an activation of the areas involved in the physiological mechanisms of sleep. PMID- 11853101 TI - Anxiogenic-like action of centrally administered glucagon-like peptide-1 in a punished drinking test. AB - A role for glucagon-like peptide-1 (GLP-1) has been postulated in the regulation of blood glucose and satiety. In addition, intracerebroventricular administration of GLP-1 has been shown to suppress locomotor activity, and produce a neuronal activation in the amygdala, a structure involved in mechanisms of fear and anxiety. Adult male Sprague-Dawley rats were prepared with chronic intracerebroventricular cannulae. Measures of experimental anxiety were assessed by the Vogel conflict test and the elevated plus maze. Central GLP-1 (fragment 7 36) administration produces a proconflict effect in the punished drinking test, while leaving measures of activity and nociception unaffected. GLP-1 may participate in the control of fear-induced suppression of behavior, probably via action in the amygdala. PMID- 11853102 TI - Auditory event-related potentials in panic and generalised anxiety disorders. AB - In this study, we aimed to investigate event-related potential (ERP) changes in panic disorder (PD) and generalised anxiety disorder (GAD) and to determine whether two disorders are different from each other in terms of endogenous potentials. A total of 35 outpatients who fully met DSM-III-R criteria for PD (8 males and 27 females) were included in this study as the PD group. The GAD group consisted of 30 subjects (5 males and 25 females) who met DSM-III-R GAD criteria. The control group consisted of 29 healthy age and sex-matched volunteers (5 males and 24 females) having no history of psychiatric or neurological illness. ERPs were recorded by using auditory "odd-ball two-tone discrimination task" method. It was found that there was significant prolongation in P3 latency in the PD group compared to the GAD and control groups. Our study suggests that there are some disturbances in early information processing in patients with PD but not with GAD. PMID- 11853103 TI - Alteration of glutamate receptors in the striatum of dyskinetic 1-methyl-4-phenyl 1,2,3,6-tetrahydropyridine-treated monkeys following dopamine agonist treatment. AB - The effects of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced nigrostriatal lesion and dopaminomimetic treatment on parameters of glutamatergic activity within the basal ganglia of monkeys were studied in relation with the development of dyskinesias. Drug-naive controls, saline-treated MPTP monkeys, as well as MPTP monkeys treated with either a long-acting D2 agonist (cabergoline) or a D1 agonist (SKF-82958) given by intermittent injections or continuous infusion, were included in this study. 3H-L-glutamate, 3H-alpha-amino-3-hydroxy-5 methylisoxasole-4-propionate (AMPA), 3H-glycine, 3H-CGP39653 (an N-methyl-D aspartate, NMDA, antagonist selective for NR1/NR2A assembly) and 3H-Ro 25-6981 (an NMDA antagonist selective for NR1/NR2B assembly), specific binding to glutamate receptors, the expression of the NR1 subunit of NMDA receptors and glutamate, glutamine and glycine concentrations were studied by autoradiography, in situ hybridization and high-performance liquid chromatography (HPLC), respectively. Pulsatile SKF-82958 and cabergoline treatment relieved parkinsonian symptoms, whereas animals continuously treated with SKF-82958 remained akinetic. Pulsatile SKF-82958 induced dyskinesias in two of the three animals tested, whereas cabergoline did not. MPTP induced no significant changes of striatal specific binding of the radioligands used, NR1 mRNA expression and amino acid concentrations. In the putamen, pulsatile SKF-82958 treatment was associated with decreased content of glycine and glutamate, whereas only glycine was decreased in cabergoline-treated monkeys. Cabergoline and continuous administration of SKF 82958 led to lower levels of NR1 mRNA in the caudate in comparison to pulsatile SKF-82958 administration. The development of dyskinesias following a D1 agonist treatment was associated with an upregulation of 3H-glutamate [+49%], 3H-AMPA [+38%], 3H-CGP39653 [+ 111%], 3H-glycine [+ 26%, nonsignificant] and 3H-Ro 25 6981 [+ 33%] specific binding in the striatum in comparison to nondyskinetic MPTP monkeys. Our data suggest that supersensitivity to glutamatergic input in the striatum might play a role in the pathogenesis of dopaminomimetic-induced dyskinesias and further support the therapeutic potential of glutamate antagonists in Parkinson's disease. PMID- 11853104 TI - A retrospective study of clozapine and electroencephalographic abnormalities in schizophrenic patients. AB - This study investigated the incidence and nature of clozapine-associated electroencephalographic (EEG) abnormalities and the relationship between EEG abnormality and clozapine dosage in Korean schizophrenic patients. Fifty schizophrenic patients with normal baseline EEG and with additional EEG record examined during clozapine treatment more than once were included. Thirty-one patients (62%) showed abnormal EEGs after clozapine treatment, and two of them had seizures. The majority of EEG abnormalities presented as nonspecific slow waves (SW). Spikes (or spike and wave complexes; SP) and frontal intermittent rhythmic delta activity (FIRDA) were relatively rarely observed. The probability of EEG abnormality was linearly dependent on the daily dose of clozapine and patient's age. Our results can be summarized as follows: (1) a substantial proportion of Korean patients treated with clozapine develops EEG abnormalities, and its incidence is comparable to the published results in Caucasian patients; (2) EEG abnormalities occurred in a dose-dependent manner; and (3) the occurrence of EEG abnormalities did not necessarily lead to future seizure development, except in a small number of cases. PMID- 11853105 TI - Dopamine effect on the mitochondria potential in B lymphocytes of schizophrenic patients and normal controls. AB - Brain metabolic abnormalities and aberrant dopamine (DA) metabolism have been reported in patients with schizophrenia. The authors hypothesized that mitochondria is a primary target of damage by increased free radical generation secondary to increased DA metabolism by monoamine oxidase (MAO). Epstein-Barr virus (EBV)-transformed human B-lymphocytes cell lines derived from patients with schizophrenia and normal controls were incubated in the absence or presence of DA, hydrogen peroxide (H2O2), or rotenone (Rot). The cells were then stained with rhodamine 123 (Rh 123) and analyzed for uptake using flow cytometry. Compared with untreated cells, DA significantly decreased Rh 123 uptake by the mitochondria. This effect was similar to the control cells treated with H2O2 or Rot. Nevertheless, there were no differences in Rh 123 uptake between the cells of schizophrenic patients and normal controls. This study shows that DA can impair the mitochondria membrane potential but that mechanism may not be evident in schizophrenia. PMID- 11853106 TI - Pre- and postsynaptic responses to 1-(1-naphthylpiperazine) following adaptation to stress in rats. AB - In view of a role of pre- and postsynaptic serotonin (5-hydroxytryptamine; 5-HT) receptors in adaptation to stress, effects of 1-(1-naphthylpiperazine) (1-NP) were compared in unrestrained and repeatedly restrained adapted rats. In the first part of the study, effects of various doses (1.0-15 mg/kg ip) of 1-NP were monitored on brain 5-HT metabolism (presynaptic response) and on the activity (postsynaptic response) of rats in an activity cage to which the rats were habituated before the drug administration. The drug injected at doses of 2.5-15.0 mg/kg increased motor activity and decreased brain 5-hydroxyindoleacetic acid (5 HIAA) concentration in a dose-dependent manner. In the second part of the study, rats were restrained on wire grids 2 h/day for 5 days. First-day episode of 2-h restraint decreased 24-h cumulative food intake, water intake and growth rate. The decreases attenuated following second-, third- and fourth-day episodes of 2-h restraint were not observed following fifth-day episode of 2-h restraint stress, suggesting adaptation to the stress schedule has occurred. Serotonergic and motor responses to 1-NP in unrestrained and repeatedly restrained adapted rats were compared by injecting the drug at a dose of 5 mg/kg, a dose that above results suggested would not produce maximal effects on 5-HT metabolism or motor activity. Administration of 1-NP at a dose of 5 mg/kg increased motor activity and decreased brain 5-HIAA concentration in unrestrained and repeatedly restrained adapted rats. Increases of motor activity were much greater in repeatedly restrained adapted than unrestrained rats. Decreases of 5-HIAA concentration were comparable in the two groups. The results are discussed in the context of an increase in the effectiveness of postsynaptic 5-HT-1A and 5-HT-1B receptors and a decrease in the effectiveness of presynaptic 5-HT-1A (somatodendritic) and 5-HT 1B (terminal) receptors following adaptation to stress. It is suggested that these changes of receptor responsiveness might help coping with stress demand to produce adaptation to stress. PMID- 11853107 TI - Electrophysiological indication for a link between serotonergic neurotransmission and personality in alcoholism. AB - The serotonergic neurotransmission seems to be involved in the neuropsychobiology of alcoholism. The intensity dependence of the N1/P2 component of auditory-evoked potentials is discussed as an indicator of the central serotonergic neurotransmission in healthy subjects. The aim of this study was to verify this correlation between intensity dependence and serotonergic neurotransmission, as indicated by the personality trait "harm avoidance" (HA) within the Temperament and Character Inventory (TCI) in alcohol-dependent patients. The intensity dependence was measured in 25 alcohol-dependent patients, 10 and 40 days after detoxification. The personality trait HA was assessed, which is supposed to reflect the serotonergic neurotransmission. The intensity dependence was negatively correlated with the temperament trait HA of the TCI (r = - .55, P<.01) at Day 40, but neither on Day 10 nor with the other personality dimensions. We conclude, that the intensity dependence reflects the serotonergic neurotransmission in withdrawn alcohol-dependent patients. PMID- 11853108 TI - Expression of nitric oxide synthases in the anterior horn cells of amyotrophic lateral sclerosis. AB - The authors investigated for a correlation between the expression of nitric oxide synthases (NOSs) with the severity of motor neuronal loss in the anterior horns of patients with amyotrophic lateral sclerosis (ALS). Spinal cords from six patients with ALS and from three normal controls were examined. The sections of cervical, lumbar, and sacral cord including Onuf's nucleus, which are seldom degenerated until the late stage, were stained with three antibodies against NOSs (anti-n-NOS, anti-e-NOS, and anti-i-NOS) using ABC methods. Perikarya of motor neurons in ALS, but not in controls, were immunoreactive against anti-n-NOS and e NOS. Anti-i-NOS did not recognize the motor neurons of ALS or of controls. The immunoreactivity for n- and e-NOSs was approximately the same in the sections of cervical, lumbar, and sacral cord in ALS. No significant differences in immunoreactivity were observed among the patients with ALS. These results suggest that the expression of NOSs does not immediately affect neuronal loss in ALS. PMID- 11853109 TI - Effects of rotational side preferences on immobile behavior of normal mice in the forced swimming test. AB - It has been suggested that side preferences in spontaneous rotational behavior are determinant of differences in vulnerability to the effects of the learned helplessness paradigm. The purpose of the present study was to investigate the effects of side preferences of rotational behavior in another animal model of depression, the forced swimming test. Immobility was also investigated upon repeated testing sessions and in interaction with sex. Swiss mice (69 males and 73 females) were submitted to three sessions (test time = 5 min) of forced swimming. Immobile and turning behaviors were measured for each session and within each testing session. Consistency of laterality was defined considering the persistence of the same side turning preference in the three sessions. In general, there was an increase in immobility as test progressed and upon repeated testing sessions. Marked interindividual differences in mice immobile behavior were observed when consistency of laterality was considered. Consistent-right turners presented greater immobility in the first session and better test-retest reliability, indicating that for this group, the adoption of immobile behavior was faster and more reliable over time. Immobility was higher for side-consistent males than for side-consistent females in the first session. This difference became even greater when consistent-right-turner males were compared to consistent-left-turner females. These results reinforce the idea that side preferences of spontaneous rotational behavior may account for interindividual differences in animal models of depression. PMID- 11853111 TI - The effect of nitric oxide synthesis inhibition on intravenous cocaine self administration. AB - Adult male rats were implanted with intravenous catheters. After a minimum of 10 days recovery from surgery, rats were trained to intravenously self-administer cocaine (1 mg/kg/infusion) during 3-h test sessions. The nitric oxide synthase (NOS) inhibitor Nomega-nitro-L-arginine methyl ester hydrochloride (L-NAME) was used to determine the effect of nitric oxide (NO) synthesis inhibition on cocaine self-administration. A 5-day protocol was used and on Days 2 and 5, an intraperitoneal injection of L-NAME (0, 3, 30, 300 mg/kg) was administered 45 to 60 min into a 3-h test session. One to two hours following L-NAME administration, there was a dose-dependent decrease in the amount of self-administered cocaine and an increase in the interresponse time (IRT) between successive cocaine injections. L-NAME appeared to prolong the rewarding effect of cocaine possibly through a pharmacokinetic action. PMID- 11853110 TI - A survey of prescribing practices in the treatment of depression. AB - BACKGROUND: With the increasing number and type of antidepressants available to clinicians, there is a need to better understand current prescribing practices and to what degree these practices reflect research findings. The purpose of this study was to examine prescribing practices in a sample of psychiatrists attending a psychopharmacology review course and compare these results with empirical evidence. METHOD: 439 of 800 clinicians asked (55%) responded to a 10-item questionnaire that was given prior to beginning the review course. Items covered three major content areas: first-line preferences in the treatment of depression, antidepressant agents most associated with certain side effects, and first-line preferences in the treatment of certain depressive subtypes. RESULTS: 214 (49%) clinicians indicated a belief that one antidepressant type is more efficacious than others. Of these 214 clinicians, 103 (48%) indicated selective serotonin reuptake inhibitors (SSRIs) as being most efficacious, while 53 (25%) indicated venlafaxine as being most efficacious; 378 (93%) clinicians indicated SSRIs as their first-line treatment preference. Mirtazapine (56%) was endorsed as most likely to be associated with weight gain, fluoxetine (57%) with sexual dysfunction, paroxetine (48%) with a discontinuation syndrome, and fluoxetine (52%) with agitation. For the treatment of anxious, atypical, and melancholic depression, SSRIs were the first choice of treatment (58%, 57%, and 57%), and for depression with prominent insomnia, mirtazapine and nefazadone (31% and 27%) were the first choices of treatment. CONCLUSIONS: Despite the lack of evidence of a significant difference in efficacy between older and newer agents, clinicians perceive the newer agents to be more efficacious than the older drugs [tricyclic antidepressants (TCAs) and monoamine oxidase inhibitors (MAOIs)] even in the melancholic and anxious depressive subtypes. Similarly, although sexual dysfunction and agitation appear to occur at similar rates with all the SSRIs, fluoxetine was perceived to be most likely to cause these side effects. These findings are significant as they highlight the discrepancy between empirical evidence and clinical practices and suggest that other factors influence clinicians' medication choices in the treatment of depression. PMID- 11853112 TI - Treatment of alcohol withdrawal with gabapentin. AB - Gabapentin is an anticonvulsant agent, also effective in the treatment of mood disorders and anxiety disorders. Three cases of alcohol withdrawal treated with gabapentin are presented. All patients received gabapentin 400 mg tid for 3 days, 400 mg bid for 1 day, and finally 400 mg for 1 day. Withdrawal symptoms subsided and no adverse effects were observed. The possible effectiveness of gabapentin in the treatment of alcohol withdrawal warrants further investigation by systematic and well-designed studies. PMID- 11853113 TI - Improvement of both psychotic symptoms and Parkinsonism in a case of dementia with Lewy bodies by the combination therapy of risperidone and L-DOPA. AB - A 69-year-old female of dementia with Lewy bodies (DLB) with fluctuating cognitive impairment, visual and auditory hallucinations, persecutory delusions and Parkinsonism was treated by the combination of 1 mg/day risperidone and 300 750 mg/day L-DOPA. By this combination therapy, both the psychotic symptoms and Parkinsonism improved, while cognitive function did not deteriorate. This report suggests that the combination therapy of risperidone and L-DOPA may be worth considering in the special cases of DLB. PMID- 11853114 TI - Treatment of social anxiety disorder with citalopram. AB - Selective serotonin reuptake inhibitors (SSRIs) are used as treatment for generalized and specific social phobias (social anxiety disorders). The efficacy of citalopram, an SSRI, for the treatment of social anxiety disorders has not yet been fully evaluated. These cases suggest that citalopram may be an effective treatment for social anxiety disorder (social phobia). These cases are consistent with two other published reports with citalopram from outside the US. Randomized controlled studies are warranted. PMID- 11853116 TI - Cytokine profiles in schizophrenic patients treated with risperidone: a 3-month follow-up study. AB - An increasing body of evidence suggests a role for the immune system in the pathogenesis of schizophrenia. The information concerning the effects of antipsychotics on cytokine profiles are limited and often controversial in particular regarding novel antipsychotics. The authors first investigated the production of various cytokines [interleukin (IL)-2, IL-4, IL-10, interferon (INF)-gamma] in drug-free (n = 12) and drug-naive (n = 3) schizophrenic patients and in healthy controls (n = 33) and then the modifications of cytokines values during a 3-month period of treatment with risperidone. In the baseline condition, the production of IL-2 and INF-gamma was significantly higher (P = .023 and .026, respectively) in patients than in controls. In the same patients, the use of risperidone was associated with augmented IL-10 (a suppressor of Type I cytokines) and decreased INF-gamma production. This modification suggests that clinical improvement is associated with a reduction in the inflammatory-like situation present in not currently treated schizophrenic patients. PMID- 11853117 TI - Calcitonin gene-related peptide and calcitonin in the CSF of patients with dementia and depression: possible disease markers. AB - Cerebrospinal fluid (CSF) was obtained from 32 patients with dementia, 19 healthy controls that were age-matched with the dementia patients, and 29 DSM-IV major depression patients and calcitonin gene-related peptide-like immunoreactivity (CGRP-LI) and calcitonin-like immunoreactivity (CT-LI) measured by RIA. CGRP-LI was lower in the dementia group compared to both the controls and depressed patients (P<.01) after covarying out sex and age. CT-LI was decreased in the dementia and depressed patients (P<.05) compared to the controls. A positive relationship between CGRP-LI and CT-LI was found in dementia. A logistic discriminant analysis with calcitonin gene-related peptide (CGRP) and log calcitonin (CT) predicting diagnosis (three classes) revealed a significant overall fit (chi2 = 18.08, P = .0011), with an effect test showing contributions of both independent variables: CGRP (chi2 = 10.03, P<.007), log CT (chi2 = 8.63, P = .013). In dementia, both CGRP-LI and CT-LI were decreased and their concentration ratio did not differ from that in controls, likely reflecting a general neuronal loss. Alternatively and more speculatively, but theoretically possible, expression of the alpha-CGRP/CT gene may be affected in dementia. In contrast, in depression, CT-LI but not CGRP-LI was decreased and the CGRP/CT concentration ratio was increased, which is consistent with a possibility of an altered splicing process favoring CGRP mRNA. PMID- 11853115 TI - Defining the neuromolecular action of myo-inositol: application to obsessive compulsive disorder. AB - Dietary inositol is incorporated into neuronal cell membranes as inositol phospholipids where it serves as a key metabolic precursor in G protein-coupled receptors. In the brain, several subtypes of adrenergic, cholinergic, serotonergic and metabotropic glutamatergic receptors are coupled to the hydrolysis of phosphoinositides (PI) with myo-inositol (MI) crucial to the resynthesis of PI and the maintenance and effectiveness of signalling. Despite a mode of action that remains illusive, MI has demonstrated therapeutic efficacy in obsessive-compulsive disorder (OCD), putative OCD-spectrum disorders, as well as panic and depression. Behavioural and biochemical studies indicate that this efficacy does not involve simply the replenishing of the membrane PI pool. In addition to its precursory role in cell signalling, inositol lipids alter receptor sensitivity, can direct membrane trafficking events, and have been found to modulate an increasing array of signalling proteins. These effects may afford MI an ability to modulate the interaction between neurotransmitters, drugs, receptors and signalling proteins. This paper reviews the neuromolecular and genetic aspects of OCD in terms of the PI-linked 5HT receptor subtypes and relates these to the behavioural and therapeutic effects of MI. Since OCD often is poorly responsive to current drug treatment, understanding the neuropharmacology of MI holds great promise for understanding the neuropathology of this and other MI-responsive disorders. PMID- 11853118 TI - A volumetric biochemical niacin flush-based index that noninvasively detects fatty acid deficiency in schizophrenia. AB - (1) It is possible to investigate aspects of phospholipid-related signal transduction in humans noninvasively using the niacin skin flush test. (2) Patients with schizophrenia have previously been reported to show a reduced flushing response. (3) The aim of this study was to devise a comprehensive index of cutaneous response to the niacin test, incorporating aqueous methyl nicotinate concentration and time, and to test this index in schizophrenia. (4) A discrete approximation to a continuous volumetric index, which we have named the volumetric niacin response (VNR), was devised. Its value was measured in 27 patients with DSM-IV schizophrenia and 26 age- and sex-matched normal controls. (5) The mean value of the VNR in the patients with schizophrenia (16.26) was significantly smaller than that of 26.77 in the normal controls (P<.0004). (6) With a threshold value for the VNR of 21, the test differentiated well between schizophrenia and normal controls (P=.002) with a sensitivity of 78% and a specificity of 65%. (7) The present results confirm that the flushing response is reduced in schizophrenia, and show that calculation of the VNR is an effective means of allowing the total response in different patients or patient groups to be readily compared. PMID- 11853119 TI - Therapeutic effects of bromperidol on the five dimensions of schizophrenic symptoms. AB - Therapeutic profiles of bromperidol and their relationship with plasma drug concentration and prolactin response were investigated in 30 acutely exacerbated schizophrenic patients treated with randomly allocated fixed-doses of bromperidol (6, 12 or 18 mg/day) for 3 weeks. The mean values (+/- S.D.) of percentage improvement at 3 weeks in total Brief Psychiatric Rating Scale (BPRS) and five subgrouped symptoms were 59.8+/-29.2% for total, 64.6+/-37.5% for positive, 73.3+/-33.7% for excitement, 80.2+/-45.5% for cognitive, 43.1+/-46.5% for negative and 49.6+/-46.8% for anxiety-depression symptoms, respectively. Twenty (67%) of 30 patients were responders defined as having 50% or more symptom reduction significantly greater in responders than those in nonresponders after 2 weeks. Mean plasma bromperidol concentration in patients with 50% or more reduction in positive symptoms was significantly higher than in the others (8.2+/ 4.7 vs. 4.1+/-1.8 ng/ml, P< .05). Percentage improvement in total BPRS at 1 and 2 weeks were correlated well with that at 3 weeks. These findings suggest that an early improvement in positive and anxiety-depression symptoms results in favorable outcome of total response to bromperidol treatment. Plasma drug monitoring may have a limited predictive value for improvement in positive symptoms. PMID- 11853120 TI - Brain lesions induced by chronic cocaine administration to rats. AB - Cocaine is a common drug of abuse, and its use has emerged as a major public health problem with neurological complications. In this work, the authors studied microscopic lesions produced in brain by chronic cocaine administration to rats. Twenty-five Wistar rats were exposed to 30 mg/kg/day ip of cocaine and sacrificed at 15, 30, 45, 60, and 90 days after treatment and compared to 25 control rats injected daily with saline. The parietal cortex (Cx), hippocampus (Hp), substantia nigra (SN), and cerebellum (Ce) were morphologically analyzed. The authors found progressive light microscopic lesions in all regions studied, including nuclear pyknosis and atrophy, interstitial edema, broken fibers, and necrosis. Results show that chronic treatment with cocaine in rats leads to selective severe lesions in different brain regions. PMID- 11853121 TI - Cannabinoid-induced stimulation of motor activity in planaria through an opioid receptor-mediated mechanism. AB - Planaria, the most primitive example of centralization and cephalization of the nervous system along phylogeny, shows specific stereotyped behavioral patterns following exposure to drugs acting on neural transmission. In this study, the authors investigated the effects of exposure to the synthetic cannabinoid receptor agonist WTN55212.2 on motor activity in planaria. WTN55212.2 produced dose-dependent stimulation of motor behavior. High doses of the drug caused stereotyped activities identical to those seen previously with opioid agonists. These effects were antagonized by coexposure to cannabinoid or opioid receptor antagonists. The results indicate that functional interactions between cannabinoid and opioid systems are highly conserved along phylogeny, at least at the behavioral level. PMID- 11853122 TI - Coadministration of L-NOARG and tiapride: effects on catalepsy in male mice. AB - This study was designed to determine the possible potentiation of catalepsy behavior after coadministration of N(G)-nitro-L-arginine (L-NOARG), an inhibitor of nitric oxide synthase (NOS), and tiapride, a specific antagonist for D2 receptors, in male mice. Catalepsy was measured by the bar test. Two successive evaluations were carried out 60 and 90 min after injections. The induction of catalepsy following coadministration of L-NOARG (25 and 50 mg/kg) and tiapride (200 mg/kg) was significantly higher than the sum of catalepsy scores after administration of L-NOARG and tiapride separately. Coadministration of L-NOARG and tiapride produced a clear potentiation of their effects on catalepsy in mice. These results underline the view that nitric oxide (NO) interacts with central dopamine D2 transmission. PMID- 11853123 TI - Comparative effects of dehydropirlindole and other compounds on rat brain monoamine oxidase type A. AB - Dehydropirlindole (DHP) is the dehydroderivative of pirlindole, a short-acting inhibitor of monoamine oxidase type A (MAO-A). DHP would be formed in vivo from oxidation of pirlindole by MAO-A. The aim of this work is to compare the inhibitory potency of DHP with three reference compounds: harmaline, befloxatone and clorgyline; the two former are reversible inhibitors and the later is an irreversible inhibitor of MAO-A. Both in vitro and ex vivo assays were performed on rat brain homogenates, and IC50 and ID50 were calculated by a fluorometric method with octopamine as selective MAO-A substrate. In vitro clorgyline and befloxatone were more potent inhibitors than DHP and harmaline with IC50 values of 1.6 and 7.7 nM vs. 40 and 55 nM; ex vivo ID50 values were 1.5 and 32 micromol/kg vs. 41 and 49 micromol/kg. Befloxatone had an ID50/IC50 ratio four to five times higher than DHP and harmaline. Preincubation time experiments did not distinguish befloxatone from DHP and harmaline. In conclusion, this study shows that DHP behaves as a reversible MAO-A inhibitor whose potency is situated between that of befloxatone and harmaline. PMID- 11853124 TI - ATPases enzyme activities during ageing in different types of somatic and synaptic plasma membranes from rat frontal cerebral cortex. AB - The catalytic properties of energy-utilizing ATPases enzyme systems related to ions homeostasis were evaluated in different types of synaptic plasma membranes (SPM) and in somatic plasma membranes (SM) from cerebral cortex of rats aged 5, 10, and 22 months. The following enzymes were evaluated: Na+, K+-ATPase, Ca2+, Mg2+-ATPase, Mg2+-ATPase and the activity of acetylcholine esterase (AChE) was also evaluated. The ATPases located on SM and SPM and synaptic vesicles are involved in the regulation of presynaptic nerve ending homeostasis and postsynaptic activities. Different types of SM and SPM (three types) were obtained by combinations of differential and density gradient ultracentrifugation techniques in sucrose-Ficoll media: the first was obtained by purification of the sediment of mitochondrial supernate and the second after synaptosomal lysis and purification on density gradient. In the cerebral cortex of 5-month-old rats, the catalytic properties of ATPases systems markedly differ according to the different types of SPM and SM, thus indicating that the metabolic role of each ATPase is determined by their subcellular in vivo localization. As regards ageing: (i) ATPase enzyme catalytic activities tend to decrease during ageing in a complex way; (ii) ageing induced specific modifications in individual ATPases according to their subsynaptic localization; and (iii) these effects are probably due to specific biochemical situations that take place at each age, reflecting the bioenergetic state of the cerebral tissue with respect to the energy demand. The cerebral concentration and content of SM proteins were increased by ageing suggesting that many defective noncatalytic proteins may be formed during ageing, as shown by immunoblotting techniques. PMID- 11853125 TI - Neuroleptic-like profile of the cannabinoid agonist, HU 210, on rodent behavioural models. AB - (1) The present study was performed to assess the effects exerted by the cannabinoid (CB) agonist, (-)11-hydroxy-delta8-tetrahydrocannabinol dymethylheptyl (HU 210; 12.5-50 microg/kg ip), on rodent behavioural tests involving dopamine (DA) transmission; in comparison, the DA D2 antagonist, S(-)-3 chloro-5-ethyl-N-[(1-ethyl-2-pyrrolidinyl)methyl]-6-hydroxy-2-methoxy-benzamide hydrochloride ((-)eticlopride; 50 microg/kg sc), was used. (2) In rats, HU 210, at all doses, potently antagonized penile erection (PE) and stretching-yawning (SY) typically elicited by the DA D2/D3 agonists, 6-allyl-2-amino-5,6,7,8 tetrahydro-4H-thiazolo-[4,5-d]-azepine (B-HT 920) and +/-7-hydroxy-N,N-di-n propylaminotetralin hydrobromide (7-OH-DPAT) both at 100 pg/kg ip. (3) In nonreserpinized mice, HU 210 impaired motor ability assessed by means of a motor test battery, and B-HT 920 (1 mg/kg ip) worsened the phenomenon. (4) In reserpinized mice, HU 210 at 50 microg/kg counteracted the amelioration exerted by B-HT 920 (1 mg/kg ip) on reserpine-induced akinesia. (5) As all these effects were similarly displayed by (-)eticlopride (50 microg/kg sc), our data suggest a neuroleptic-like profile of acute HU 210 in animal behavioural tests. PMID- 11853126 TI - Insulin-like growth factor-1 does not ameliorate the age-related decline in presumptive inhibitory synapses in layer 2 of rat sensorimotor cortex. AB - Four old (29 months) Brown Norway x Fischer 344 (BN x F344) rats received intracerebroventricular infusion of insulin-like growth factor-1 (IGF-1) and four middle-aged (18 months) and four old (29 months) rats received infusion of saline for 28 days. Sensorimotor cortex containing layer 2 was blocked and processed for electron microscopy. Thin (700 A) and semithin (1 microm) sections were collected from the same anatomical space for quantification of synapses and neurons, respectively, using the physical disector. Numerical density (Nv) of presumptive inhibitory synapses in layer 2 of sensorimotor cortex has been reported to decline with age Poe et al., 2001; Brunso-Bechtold et al. [Brain Res. 872 (2000) 125]. Infusion of IGF-1 did not affect the density of synapses or neurons when old IGF-1 animals were compared with old saline animals. PMID- 11853127 TI - Psychological distress in childhood trauma survivors who abuse drugs. AB - The relationships between the level of childhood maltreatment and current psychological distress were examined in a community sample of 676 substance abusing men and women using a validated self-report instrument (the Childhood Trauma Questionnaire) designed to measure physical, emotional, and sexual abuse and physical and emotional neglect. Current levels of self-reported psychological distress/symptoms were measured using a 53-item Brief Symptom Inventory. Prevalence of early trauma ranged from 44% for emotional neglect to 65% for sexual abuse. The severity of all forms of childhood maltreatment were directly associated with current psychological distress. PMID- 11853128 TI - Early life sexual abuse as a risk factor for crack cocaine use in a sample of community-recruited women at high risk for illicit drug use. AB - Subjects for this study were 1,478 community-recruited women sexual partners of male injection drug users who were participants in the National Institute on Drug Abuse (NIDA)-supported Women Helping to Empower and Enhance Lives (WHEEL) project. This study assessed the association between child/adolescent sexual abuse--including specific type of abuse and perpetrator of abuse--and lifetime crack use in this sample of women. About 64% of sample women had ever used crack; 56% had been sexually abused by age 18. In logistic regression analyses, any sexual abuse in childhood, penetrative sexual abuse in childhood, and sexual abuse by a family member in childhood were significantly associated with lifetime crack use. Sexual abuse in adolescence was indirectly associated with lifetime crack use through running away from home and rape in adulthood. Given that many of these subjects reported drug treatment experience, such programs may provide the best setting for helping women with both substance use and sexual abuse issues. PMID- 11853129 TI - A synthesis of welfare reform policy and its impact on substance users. AB - OBJECTIVES: The purpose of this study was to provide an overview of welfare reform and its impact on the substance-abusing recipient. METHODS: The data for this paper were derived from sources including the US Department of Health and Human Services and the National Institute on Drug Abuse. RESULTS: The number of individuals on public assistance has decreased in the years following implementation of welfare reform legislation. Factors relevant to transitioning welfare recipients into the workplace, such as transportation and childcare, have special ramifications for the drug-using population. Additionally, these individuals require treatment for their addictions in order to become employable. The issue of concern is that recipients may be deterred from seeking benefits by various provisions of welfare reform legislation and turn instead to other sources (including illicit activities) for sustenance. CONCLUSIONS: Welfare caseloads have been dropping over the past two years. However, the number of substance abusers continues to rise. It is not known in what ways welfare reform will affect substance abusers who are welfare recipients. Important policy issues arise from this nexus; it is argued that these issues will require careful investigation. PMID- 11853130 TI - Predictive validity of a measure of treatment readiness for out-of-treatment drug users: enhancing prediction beyond demographic and drug history variables. AB - The predictive validity of a treatment readiness measure designed for out-of treatment drug users, tapping dimensions of perceived problem severity, perceived need for formal treatment, motivation for treatment, and negative attitudes toward treatment is examined using data from a National Institute on Drug Abuse (NIDA)-funded HIV outreach intervention in San Antonio, Texas. Logistic regression is used to predict use of substance abuse treatment services three months after program intake on the basis of readiness scores, controlling for demographic and drug history variables. Analyses of data from 673 drug users in an HIV outreach intervention indicated that treatment readiness dimensions accounted for a 12% increase in variation in "use of any modality" and 14% for "use or attempted service use". Only Motivation to Quit was a significant predictor of "use of any modality" and both Perceived Need for Treatment and Motivation to Quit were significant predictors of "use or attempted use". Findings support the importance of the individual's perception of "readiness" to change their substance-abusing behavior and enter treatment. PMID- 11853131 TI - The relationship between the consumption of various drugs by adolescents and their academic achievement. AB - This study uses the NELS data set to assess the relationship between adolescent patterns of the consumption of marijuana, cocaine, alcohol, and cigarette consumption and their academic achievement. The results of the study indicate that increased frequency of cigarette smoking and being under the influence of marijuana, cocaine, and alcohol did frequently have an impact on adolescent academic achievement. When all types of drug consumption were considered together, cigarette smoking, being drunk, and under the influence of alcohol while at school were the variables that produced the most consistent statistically significant effects. Being under the influence of cocaine generally produced the largest regression coefficients, but the effects were often not statistically significant, which may in part be due to the small portion of students who reported being under the influence of cocaine while at school. When examined separately, being under the influence of cocaine and marijuana each had consistently statistically significant effects on academic achievement. PMID- 11853132 TI - Inhalant abuse among American Indian, Mexican American, and non-Latino white adolescents. AB - The abuse of volatile solvents, or inhalants, is an enduring problem among adolescents although a number of factors obscure the nature and extent of this drug using behavior. The data presented here indicate that a number of social and perceptual correlates of inhalant use operate similarly across Mexican American, American Indian and non-Latino white adolescents. Peer factors appear to dominate, although they are somewhat less important for Mexican American and Indian youth. Increased perception of harm reduces inhalant use for all groups. Of particular significance in the findings here are the continued increase of inhalant use among females compared to males and the strong pattern of decreases in inhalant use among American Indian adolescents over the last decade. A number of implications for increased effectiveness of prevention are discussed. PMID- 11853133 TI - Open-label pilot study of bupropion plus bromocriptine for treatment of cocaine dependence. AB - Combinations of medications are often used in neuropsychiatry to enhance treatment efficacy. This 8-week, open-label study tested the combination of bupropion (< or =300 mg) and bromocriptine (< or =7.5 mg) daily in 34 cocaine dependent (DSM-IIIR) outpatients also receiving weekly individual counseling. The first 18 subjects spent one week at maximum dose; the next 16 spent three weeks. Both groups showed significant reductions in self-reported cocaine use, with no significant change in proportion of urine toxicology tests positive for cocaine. There were no significant differences in outcome between groups. These results suggest that the combination of bupropion and bromocriptine is safe in cocaine addicts, but provide ambiguous evidence of its efficacy. PMID- 11853134 TI - Substance use and labor force participation among homeless adults. AB - OBJECTIVES: We measured the proportion of homeless adults in the labor force and examined the impact of substance use on labor force participation. METHODS: A county-wide probability sample of 397 homeless adults was interviewed three times in a 15-month period. RESULTS: Almost 80% of homeless adults were employed or looking for work at one point in time; however, only 47.7% remained in the labor force over the 15-month study period. Recent drug users were only 5% as likely as other homeless adults to be in the labor force; and consistent public entitlement recipients were only 18% as likely as other homeless adults to be in the labor force. CONCLUSIONS: Recent illicit drug use posed a deterrent to labor force participation among homeless adults, but heavy alcohol use did not. Most homeless adults were not consistently in the labor force and those who were, did not receive public entitlement benefits. This finding poses an interesting dilemma since previous studies indicated that homeless adults, who are consistent public entitlement recipients, were more likely to get housed than those who are not. PMID- 11853135 TI - Determining predictors of attrition in an outpatient substance abuse program. AB - Determining pre-treatment variables that predict attrition in an outpatient cocaine abuse program is critically important in efforts to enhance retention and ultimately improve client outcome. Potential predictors have been identified, such as treatment history, deviant behaviors, and level of drug use; however there is not widespread agreement on their applicability across treatments and populations. This study examines the relationship of demographic, drug use severity, and psychosocial factors with treatment attrition and the time of dropout. One hundred and sixty-five individuals from the Houston area, seeking treatment for cocaine dependence, completed a pre-treatment assessment battery prior to starting 12 weeks of outpatient treatment. A series of regression analyses showed that treatment dropouts were more likely to be separated from their spouses, have poorer family/social functioning, have fewer years of education, and to be female. Those participants with higher education levels and those with poorer psychiatric functioning tended to remain in treatment longer. The implications of these findings are discussed. PMID- 11853136 TI - The prevalence and clinical course of sedative-hypnotic abuse and dependence in a large cohort. AB - Relatively little is known about the prevalence and clinical characteristics of dependence on sedative-hypnotics, and almost nothing has been published regarding abuse. This report relates information on Diagnostic and Statistical Manual of the American Psychiatric Association (DSM-IIIR) sedative-hypnotic use disorders among subjects from the Collaborative Study on the Genetics of Alcoholism (COGA). A standardized interview was used to generate data on 407 men and women in Group 1 with sedative-hypnotic dependence (4.4% of the COGA sample), 34 in Group 2 with abuse (0.4%), and 3,426 comparison subjects in Group 3 with alcohol dependence in the absence of a sedative-hypnotic use disorder (36.7%). The remaining COGA subjects (48.5%) were not included as they had neither alcohol nor sedative hypnotic dependence or abuse. Those with sedative-hypnotic abuse or dependence were more likely to be Caucasian individuals with abuse or dependence on marijuana, cocaine, amphetamines, or opioids. Subjects in Groups 1 and 2 were also more likely to have histories of independent major depressive and panic disorders, as well as substance-induced mood disorders. Those with dependence, compared to abuse, were likely to be women, reported staying intoxicated for a day or more, but noted less abuse of opioids or amphetamines, although Group 2 members also had high rates of difficulties with sedative-hypnotics. These results highlight notable rates of sedative-hypnotic dependence in the COGA families, and indicate that while sedative-hypnotic abuse does occur, and while the clinical course can involve relatively serious problems, it is less common than dependence. PMID- 11853137 TI - Attitudes towards drug legalization among drug users. AB - Research shows that support for legalization of drugs varies significantly among different sociodemographic and political groups. Yet there is little research examining the degree of support for legalization of drugs among drug users. This paper examines how frequency and type of drug use affect the support for legalization of drugs after adjusting for the effects of political affiliation and sociodemographic characteristics. A sample of 188 drug users and non-drug users were asked whether they would support the legalization of marijuana, cocaine, and heroin. Respondents reported their use of marijuana, crack, cocaine, heroin, speedball, and/or methamphetamines during the previous 30 days. Support for legalization of drugs was analyzed by estimating three separate logistic regressions. The results showed that the support for the legalization of drugs depended on the definition of "drug user" and the type of drug. In general, however, the results showed that marijuana users were more likely to support legalizing marijuana, but they were less likely to support the legalization of cocaine and heroin. On the other hand, users of crack, cocaine, heroin, speedball, and/or methamphetamines were more likely to support legalizing all drugs including cocaine and heroin. PMID- 11853138 TI - Vaccine development against Neospora caninum infection. AB - Neospora caninum is a recognized protozoan parasite of a wide range of mammalian hosts, and was reported for the first time in 1988. The isolation of its oocysts in dog's faeces in 1998 led to its establishment as a parasitic species undergoing typical coccidian life cycle. Infection with N. caninum causes paralysis and death in young livestock and companion animals, and is associated with abortions and stillbirth in cattle, and neurologic disease in calves. Considering the economic and agricultural importance of neosporosis, there is the urgent need to develop biological control measures aimed at preventing its transmission, infection, as well as reducing severity of the disease. In this paper, we have reviewed the progress made to date on the parasite-host immunology and on vaccine development including its prospects, and discussed possible strategies in the formulation of vaccine(s) against neosporosis. PMID- 11853139 TI - An immunohistochemical study of endocrine cells in the abomasum of vagotomized calf. AB - The effect of thoraco-vagotomy on the distribution and frequency of chromogranin , serotonin-, somatostatin- and gastrin-immunoreactive cells in the abomasum of the calf were investigated by immunohistochemistry. Calves were vagotomized at 1 week old and sampled 2 and 4 weeks later. The endocrine cells generally decreased in number in vagotomized calves as compared to non-operated control calves. However, the detailed responses of endocrine cells to vagotomy varied depending on the endocrine cell type, region of gastric mucosa, and period after vagotomy. The present result suggests that the vagus nerve has an influence on the intrinsic regulatory system by endocrine cell control in the ruminant abomasum. PMID- 11853140 TI - Evaluation of canine gastric motility with ultrasonography. AB - For the evaluation of canine gastric motility with ultrasonography, contraction number of pyloric antrum and gastric emptying time (GET) by area and volume method developed by Bolondi et al.'s method were studied in 14 dogs. All experimental dogs were administered with saline and soup solution (10 ml/kg, B.W.). The mean values of contraction number of pyloric antrum in saline and soup group were 4.19 +/- 1.30/min and 4.82 +/- 0.65/min before feeding, and overall mean values were 4.66 +/- 1.37/min and 5.13 +/- 1.71/min, respectively. The mean values of the GET by area and volume method were 36.73 +/- 11.27, 40.00 +/- 8.87 min in saline group and 61.35 +/- 17.58, 59.11 +/- 14.46 min in soup group. In the GET in saline and soup groups, there was no significant difference between the area and volume method (p>0.05). Therefore, Bolondi et al.'s method by ultrasound can be used to evaluate the antropyloric motility and gastric emptying time with area and volume methods. The area method is easier to determine the GET than the volume method, but the latter is more accurate. PMID- 11853142 TI - Age-related changes in bone mineral density, cross-sectional area and strength at different skeletal sites in male rats. AB - Age-related changes in bone mineral density (BMD), cross-sectional area and strength strain index (SSI) of the long bones in the limbs and first lumbar vertebra of male Wistar rats were measured by a peripheral quantitative computed tomography (pQCT) method. One hundred and ten rats aged 2-30 months were used. The results indicate that the total (cortical + trabecular), cortical and trabecular BMD values of the metaphysis and cortical BMD values of the diaphysis in the long bones varied for each bone and differed from those of the first lumbar vertebra. The total BMD of long bones showed high values at 6-21 months and then decreased, but these did not always coincide with cortical and trabecular BMD. The values of SSI in the long bones varied. The values of total and cortical BMD and SSI of lumbar vertebra increased for 6-12 months and then decreased, but the trabecular BMD increased after 12 months. The total area in both the long bones and the first lumbar vertebra increased with the decrease in cortical area and the increase in the trabecular area with increasing age. It was concluded that age-related changes in bones, similar to those observed in humans, could be observed in some bones and parameters, although the age in rats when the so-called peak bone mass appears in the whole skeleton could not be clearly determined. PMID- 11853141 TI - Echo-guided studies on portal and hepatic blood in cattle. AB - The objectives of the present study were (1) to establish and maintain catheters non-surgically in the portal and hepatic veins of cattle for long-term monitoring of postprandially absorbed metabolites, and (2) to assess the safety of the technique used. Aspiration and catheterization were not performed if there was risk of bleeding, as determined by hemostatic data and enlargement of portal and hepatic veins. In five Holstein cattle, before and 60-min after feeding, percutaneous ultrasound-guided aspiration of portal and hepatic blood, together with jugular blood, was performed utilizing 17G x 17.5 cm needles. Concentration of ammonia nitrogen tended to be higher after feeding, but that of urea nitrogen and glucose were not significantly changed. In five other Holstein cows liver veins were catheterized with 14 x 16.5 cm guide-needles attached with 50 cm polyethylene tubes and blood samples were collected from portal, hepatic and jugular veins at 30-min intervals for 4.5 hr. The concentration of plasma acetate and 3-hydroxybutyrate (3-HB) were significantly increased after feeding, while that of lactate was not changed. Catheters in the portal and hepatic veins enabled the collection of blood samples up to 15 days (mean, 7 days). After a 7 day observation period, five cows were euthanised and necropsied. Changes in the intercostal musculature, peritoneum and liver were negligible. During the observation period, clinical profiles, aspartate aminotransferase, alanine aminotransferase and total and differential WBC counts were within normal ranges. PMID- 11853143 TI - Canine serum alkaline phosphatase isoenzymes detected by polyacrylamide gel disk electrophoresis. AB - Serum alkaline phosphatase (ALP) isoenzymes were studied in normal dogs using a commercially available polyacrylamide gel disk electrophoresis kit (PAG/disk kit). Serum samples taken from the dogs were incubated with neuraminidase, after which most showed ALP isoenzymes as two characteristic stained bands. To determine the origin of each band, ALP isoenzymes of serum and tissue extracts (liver, intestine and bone) were characterized by heating, wheat germ agglutinin (WGA) and levamisole treatments. The results suggested that the band detected on the anode was liver ALP (LALP) and that the band detected on the cathode represented bone ALP (BALP), and both were corticosteroid-induced ALP (CALP). The percentage of each ALP isoenzyme to total ALP activity was estimated by densitometry. The percentage of BALP was the highest in young dogs (age<1 year, 64.7% ), and this value decreased with age. In contrast, the percentage of LALP in young dogs (22.2%) was much lower than that in middle-aged dogs (ages 1 year to 7 years, 59.3%) and old dogs (ages>7 years, 50.4%). The present results suggested that a commercially available PAG/disk kit is capable of detecting three serum ALP isoenzymes in dogs, and further that it may have clinical applications in the evaluation of ALP isoenzymes in veterinary medicine. PMID- 11853144 TI - Effects of dopamine infusion on cardiac and renal blood flows in dogs. AB - In veterinary medicine, dopamine is currently being administered clinically by infusion for treatment of kidney disorders at low doses (< or = 3 microg/kg/min) and for assessment of hemodynamics at high doses (> or = 5 microg/kg/min). However, since high doses of dopamine cause peripheral vasoconstriction due to its effect on alpha adrenoceptors, high doses have no longer been recommended. The present study was conducted to explore possible regimens for the use of dopamine infusion in dogs. The regional (renal and cardiac) blood flow for 60 min was measured by using colored microspheres at three doses (3, 10 and 20 microg/kg/min) of dopamine infusion in healthy anesthetized mongrel dogs. The effects on kidney and peripheral hemodynamics at each dose and the resultant cardiac output, mean arterial blood pressure and total peripheral resistance were determined. Renal blood flow increased markedly at 3 microg/kg/min dopamine. Improvement in hemodynamics indicated by marked increase in cardiac blood flow, cardiac output and mean arterial blood pressure and decreased total peripheral resistance was observed at higher doses (10 and 20 microg/kg/min). At 10 microg/kg/min, in addition to the satisfactory increase in cardiac blood flow, there was also a stable satisfactory increase in renal blood flow. However, at 20 microg/kg/min, increased myocardial oxygen consumption (manifested by marked increased in cardiac output), arrythmia and irregular increase in renal blood flow were detected. This study suggests that the clinical use of dopamine infusion in dogs could be safely expanded to moderately higher doses. PMID- 11853145 TI - Detection of a quantitative trait locus for intramuscular fat accumulation using the OLETF rat. AB - The Otsuka Long-Evans Tokushima Fatty (OLETF) rat is an animal model for obese type 2 diabetes. We showed that the OLETF rat exhibits higher levels of intramuscular fat content in Musculus longissimus as compared to the Fischer-344 (F344) rat. Our investigation was designed to identify quantitative trait loci (QTLs) contributing to the increased levels of intramuscular fat content by performing a whole-genome search using 108 F2 intercross obtained by mating the OLETF and the F344 rats. We identified one QTL responsible for intramuscular fat accumulation on rat chromosome 1 with a maximum lod score of 3.4, which accounts for 5% of the total variance. As expected, the OLETF allele corresponds to the increased levels of intramuscular fat content. PMID- 11853146 TI - Decreases of apolipoprotein B-100 and A-I concentrations and induction of haptoglobin and serum amyloid A in nonfed calves. AB - Reduced feed intake near parturition is suggested to be one of the major causal factors for the development of fatty liver in cows, and nonfeeding has been used as an experimental model for fatty liver. In cows with fatty liver, concentrations of lipoprotein lipids and proteins are decreased. In addition, the acute-phase protein haptoglobin is induced. The purpose of the present study was to examine whether the decrease of lipoprotein concentrations and the induction of acute-phase proteins were similarly reproduced by non-feeding. Holstein female calves (n=5) were nonfed for 3 days and thereafter refed. Serum concentrations of nonesterified fatty acids and beta-hydroxybutyric acid were initially increased by the nonfeeding, and followed by decreases in concentrations of cholesteryl esters, phospholipids, apolipoprotein (apo) B-100 and apoA-I. The apoC-III concentration was not distinctly decreased. Haptoglobin and serum amyloid A were induced during the nonfeeding and refeeding process. Haptoglobin was distributed in different proportions in the high-density lipoprotein, very high-density lipoprotein and the lipoprotein-deficient fractions, whereas almost all serum amyloid A was associated with the high-density lipoprotein fraction. These results suggest that the decreases in lipoprotein concentrations and induction of acute-phase proteins found in cows with fatty liver and those with fatty liver related diseases such as ketosis are primarily due to the reduced feed intake near parturition. PMID- 11853147 TI - Postweaning multisystemic wasting syndrome of pigs in Korea: prevalence, microscopic lesions and coexisting microorganisms. AB - A retrospective study was performed on natural cases of postweaning multisystemic wasting syndrome (PMWS), recorded from January 1999 to December 2000, to determine the prevalence, microscopic lesions, and other coexisting pathogens associated with PMWS. PMWS is diagnosed based on three criteria: the presence of clinical signs (retardation of growth), characteristic microscopic lesions (granulomatous inflammation and inclusion body), and the presence of porcine circovirus (PCV)-2 within these lesions. One hundred and thirty three (8.1%) of the 1634 pigs submitted from 1243 pig farms were diagnosed for PMWS. The affected pigs were from 25 to 120 days old, the majority (78 cases, 58.6%) being 60 to 80 days old. PMWS occurred each month during the two-year study period, but the incidence peaked in May (38 cases, 28.6%), followed by April (18 cases, 13.5%) and June (13 cases, 9.8%). The most consistent and characteristic lesions were multifocal, granulomatous inflammation in lymph nodes, liver and spleen, characterized by infiltration of epithelioid macrophages and multinucleated giant cells. The majority of cases (113 cases, 85.0%) was dual infection with other pathogens. The combination of PCV-2 and Hemophilus parasuis (43 cases, 32.3%) was shown to be the most prevalent followed by PCV-2 and porcine reproductive and respiratory syndrome virus (39 cases, 29.3%). The consistent presence of PCV-2, but lower prevalence of other viral and bacterial pathogens in all pigs examined with PMWS, has led to the speculation that PCV-2 is the etiological agent causing PMWS. PMID- 11853148 TI - Successful treatment of two dogs with allergic dermatitis by anti-allergic peptides (MS-antigen). AB - The effects of non-specific immunotherapy with anti-allergic peptides extracted from the urine of human allergic patients (MS-antigen), in two dogs with allergic dermatitis (AD) have been described. Clinically, severe pruritus accompanied by secondary bacterial pyoderma did not respond to conventional therapy with systemic antibiotics. The first clinical change appeared as a significant reduction in pruritus within 3 months, around the time of the 15th injection in both cases. The clinical condition was stabilized after 5 months, allowing the gradual withdrawal of concurrent therapies and an increase of injection intervals. The correlation between the results of intradermal skin tests before and after treatment and the improvement of clinical signs was not obvious. PMID- 11853149 TI - Protective effect of Clostridium septicum alpha-toxoid vaccine against challenge with spores in guinea pigs. AB - The protective effect of an alpha-toxoid vaccine of Clostridium septicum purified alpha-toxin was investigated in guinea pigs. Purified alpha-toxin was treated with formalin to make toxoid, and alpha-toxoid vaccine was prepared by mixing alpha-toxoid (4 to 64 microg/dose) with an aluminum phosphate gel as adjuvant. Guinea pigs were immunized twice with different doses of alpha-toxoid vaccine, and challenged with spores of C. septicum. The guinea pigs surviving after challenge had been immunized with 8 microg/dose or more of alpha-toxoid. All these animals produced titers of 20 units or higher of antitoxin at the challenge. The results suggest that C. septicum alpha-toxin plays an important role in protection against challenge with spores in guinea pigs. PMID- 11853150 TI - Ocular infection of cattle with Setaria digitata. AB - One 5-month-old female native Korean calf and a 2-year-old female Holstein cow raised in two farms about 4 km apart from each other in Korea, were found to have the left eye opaque, which included motile white worms in the aqueous humor. The parasite removed from the left eye of the calf was identified as Setaria digitata based on both light and electron microscopic features. The ocular infection with S. digitata reported herein may document the first aberrant case in Korean cattle. PMID- 11853151 TI - Antimicrobial susceptibility of pathogenic Escherichia coli isolated from pigs in Korea. AB - The in vitro susceptibilities of 285 isolates of Escherichia coli from preweaned and postweaned pigs with diarrhea and edema disease were tested with the 15 commonly used antimicrobial drugs by an agar dilution minimal inhibitory concentration procedure according to National Committee for Clinical Laboratory Standards (NCCLS) guidelines. All E. coli isolates tested in this study belonged to enterotoxigenic E. coli, attaching and effacing E. coli, or Shiga toxin producing E. coli. Field isolates had low MIC90 for ceftiofur (1 microg/ml). No correlation in antimicrobial resistance was found in three types of E. coli. PMID- 11853152 TI - Purification and quantification of lactoferrin in equine seminal plasma. AB - Lactoferrin with a molecular mass of 80 kDa was purified from equine seminal plasma by heparin-Agarose affinity chromatography and Sephacryl S-200 gel filtration. Purified lactoferrin was found to be highly homogeneous on the bases of its migration as a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis and of the monospecificity of rabbit antibodies to the purified protein in immunoblotting of seminal plasma proteins. A sandwich enzyme-linked immunosorbent assay was developed for quantifying lactoferrin in equine seminal plasma. Seminal plasma lactoferrin concentrations in 23 normal stallions ranged from 42 to 453 microg/ml, with a mean value of 157 +/- 118 microg/ml (S.D.). PMID- 11853153 TI - Proliferation of canine intervertebral disk chondrocytes in three-dimensional alginate microsphere culture. AB - Proliferation of chondrocytes from nucleus pulposus (NP) and anulus fibrosus (AF) was confirmed in three-dimensional culture using alginate microspheres. Cells isolated from NP and AF were incorporated in microspheres and cultured for 14 days. Round mononuclear cells of 20-25 microm in diameter proliferated and formed aggregates. At day 14, alcian blue positive matrix surrounded the proliferating cells. The cells had cytoplasmic vacuoles stained positively by toluidine blue. On electron microscopy, the cells contained proteoglycan vacuoles and lipid droplets in the cytoplasm and synthesized collagen fibrils and electron dense granules surrounding the cell. These features of the cells were characteristic for chondrocytes. This culture system should be useful to further investigate metabolic activities of intervertebral disk chondrocytes. PMID- 11853154 TI - Induction of estrus during the non-breeding season in Egyptian Baladi goats. AB - The induction of estrus during the non-breeding season was investigated in 100 Egyptian Baladi goats (Capra hircus). All animals assigned to treatments had low progesterone concentrations (<0.5 ng/ml) tested 2 times 10 days apart to confirm anestrous condition. Animals were assigned to three experimental groups. A group of animals received subcutaneous norgestomet ear implant for 11 days and a single i.m. injection of PGF2alpha 24 hr before implant removal (group I; n=40). Second group of animals received subcutaneous norgestomet ear implant for 11 days and a single i.m. injection of PGF2alpha 24 hr before implant removal and gonadotropin releasing hormone 24 hr after implant removal (group II; n=40). Third group of animals received no treatment (control group; n=20). The percentage of goats that showed estrous behavior during the first 72 hr after implant removal was 77.5, 85.0% and 10.0% in group I, group II and control group, respectively. The fertility rate was 57.5, 70.0% and 10.0% in group I, group II and control group, respectively. In conclusion, estrus can be induced in seasonally anestrous Egyptian Baladi goats using norgestomet and PGF2alpha and the injection of GnRH 24 hr after norgestomet implant removal synchronized ovulation in a higher percentage of goats. PMID- 11853155 TI - Idiopathic thrombocytopenia in Japanese black cattle. AB - A female Japanese Black cow showed hemorrhage under the mucous membrane of the eye conjunctiva and the vagina, and thrombocytopenia in peripheral blood. The clinical manifestations repeated spontaneously during long-term observation. The clinical findings were stable except for severe periodic bleeding. Prothrombin time, activated partial thromboplastin time and fibrinogen levels were normal. The anti-platelet antibody in the serum was analyzed by indirect platelet coagulation tests. Because the platelets in the micro-plate wells of the subject series showed coagulation up to 1:128, it was judged that anti-platelet antibody was present in the serum. A number of megakaryocytes were recognized in smear samples obtained from bone marrow of the scapula immediately after euthanasia. Therefore, this case was diagnosed as idiopathic thrombocytopenia. PMID- 11853156 TI - Isolation of foot-and-mouth disease virus from Japanese black cattle in Miyazaki Prefecture, Japan, 2000. AB - Four outbreaks of foot-and-mouth disease (FMD) occurred from March to May 2000 in Miyazaki and Hokkaido Prefectures, Japan. FMD virus isolation was achieved by sampling probang materials from Japanese Black cattle in the third case found in Miyazaki Prefecture. The probang materials were inoculated to bovine kidney (BK) and bovine thyroid cell cultures. CPE was observed in the BK at two days post inoculation. Specific amplified DNA segments for FMD virus (FMDV) were detected by reverse transcriptase polymerase chain reaction in the culture fluid. The FMDV was identified as type O by enzyme-linked immunosorbent assay (ELISA) for antigen detection and the nucleotide sequence encoding the VPI was determined. This FMDV is a strain that is widespread in Pan-Asia and was designated as O/JPN/2000 by the World Reference Laboratory of the Pirbright Institute, England. This report marks the first isolation of FMDV in Japan. PMID- 11853157 TI - Gastrointestinal transit and drug absorption. AB - The gastrointestinal (GI) absorption of orally administered drugs is determined by not only the permeability of GI mucosa but also the transit rate in the GI tract. It is well known that the gastric emptying rate is an important factor affecting the plasma concentration profile of orally administered drugs, and the intestinal transit rate also has a significant influence on the drug absorption, since it determines the residence time of the drug in the absorption site. The reason why the residence time is also a critical factor for drug absorption is that there is the site difference in absorbability for some drugs. We have developed the GI-Transit-Absorption Model (GITA Model) to analyze and predict the drug absorption kinetics by taking into account both the two factors, ie. GI transit and drug absorbability including its site difference. GITA Model has been already evidenced to be very useful for estimating the absorption kinetics of drugs with various characteristics and applied to assess the human data in combination with the gamma scintigraphy. In this review, the importance of GI transit rate in determining the absorption kinetics and the bioavailability of orally administered drugs is discussed mainly employing GITA Model and the results obtained by the model. PMID- 11853158 TI - The study on a PVC membrane electrode for gemfibrozil. AB - In this paper, a poly(vinyl chloride) (PVC) membrane electrode is prepared for gemfibrozil, 2, 2-dimethyl-5-(2,5-xylyloxy) valeric acid, based on its ion pair complexes with hexadecyltrioctyl ammonium iodide (HTOA). The membrane composition of the electrode was optimized by using the sequential level elimination method for orthogonal experimental design. The electrode has a Nernstian response range from 2.5 x 10(-5) to 0.1 mol/l with an average slope of 55.3 mV/decade. The limit of detection is 7.1 x 10(-6) mol/l. The electrode responses were not affected by pH in the range 10.0-12.3. A Na2B4O7-Na2CO3 buffer of pH = 11.0 was selected as the background electrolyte solution for potentiometric measurements. The electrode was used for determining gemfibrozil in pharmaceutical preparations with satisfactory results. PMID- 11853159 TI - Application of green fluorescent protein to affinity electrophoresis; affinity of IgG-binding domain C from streptococcal protein G to mouse IgG1. AB - Affinity electrophoresis (AEP) using green fluorescent protein (GFP) was studied. We constructed a fusion protein that linked S147PGFP and IgG binding domain C from streptococcal protein G (GFP-SpGC). The affinity of GFP-SpGC for mouse IgG1 was measured. The AEP using GFP does not require a staining step after electrophoresis, and was successful with a non-purified sample. Therefore, this method is simple and useful for measuring many samples such as those used in mutational studies. PMID- 11853160 TI - Induction of CD4+ regulatory T cells by TPA in mice: contra-suppression by CD8+ T cells. AB - Among the eight inbred mouse strains employed in our preceding report, 12-O tetradecanoylphorbol 13-acetate (TPA) painting alone induced CD4+ regulatory T (Tr) cells in four strains (e.g., C3H/He) at 6-8 weeks of age, but not in the remaining strains (e.g., C57BL/6, BALB/c). In the present study, the effect of growth from 4-14 weeks on delayed-type hypersensitivity (DTH) response was investigated in three inbred murine strains, C3H/He (H-2k), C57BL/6 (H-2b) and BALB/c (H-2d) mice. In all strains older than 10 weeks, DTH response was suppressed exclusively by TPA painting. The defect of suppressive activity for DTH in several of the strains at 6-8 weeks of age was dependent on the presence of cells, which blocked regulatory cell activity at 6-8 weeks of age, but not at 10 weeks of age. The age-dependent difference in regulatory activity was caused by the presence of CD8+ contra-regulatory T (Tcr) cells. CD8+ contra-regulatory T cells are required to contact regulatory cells in order to block DTH suppressive activity. Adhesion molecules were of great importance in contra-suppression, as antibody treatment to LFA-1 or ICAM-1 blocked this activity. ICAM-1 expression on CD4 T cells greatly increased following growth in 10 week-BALB/c mice receiving TPA than 6-week-old mice, however, a slight increase in growth occurred in 6-to 10-week-old animals in which TPA was absent. The degree of increment in body weight was very similar in these inbred strains. Thymus involution in C3H/He mice was the earliest signal among these mice. This result may suggest that the period of differentiation and maturation of T cells in a first lymphoid tissue for the growth process differs in these three inbred strains. This study provides an interesting example of genetic control of maturation or proliferation of peripheral T cells. PMID- 11853161 TI - Effect of protein phosphatase inhibitors on the development of mouse embryos: protein phosphorylation is involved in the E-cadherin distribution in mouse two cell embryos. AB - Protein phosphorylation plays many important roles in cell functions and cell differentiation. To clarify the roles of protein phosphorylation in early embryonic development in mice, 2-cell embryos were cultured in the presence of various protein phosphatase inhibitors such as calyculin A, okadaic acid, cyclosporin A, tacrolimus (FK506) and benzyl-phosphonic acid. Calyculin A potently inhibited the 2-cell cleavage to the 4-cell stage. The concentration for 50% inhibition was 0.26 nM. At the same time, we found that calyculin A-treated 2 cell embryos showed a morula-like shape at a concentration of 2 nM in 24 h. It is well known that E-cadherin plays a key role in the compaction of late 8-cell stage embryos. In this report, we observed the distribution of E-cadherin protein using anti-E-cadherin antibody with a fluorescence microscope, and also evaluated the relative E-cadherin mRNA content at various stages of embryos by RT-PCR and ABI PRISM 7700 System (a real time PCR apparatus). The fluorescence intensity of E-cadherin increased along with the embryonic development. During the embryonic development from the 2-cell stage to the blastocyst stage, the relative E cadherin mRNA content greatly increased in a time-dependent manner, while the mRNA did not increase with the addition of calyculin A at the 2-cell stage. Therefore, we observed the localization of the E-cadherin protein in calyculin A treated embryos with a laser microscope. The distribution pattern of E-cadherin was altered by the addition of calyculin A from a scattered pattern throughout the embryos to a localized pattern at the cell-cell boundary region. These results strongly suggest that the distribution of E-cadherin protein is regulated by protein phosphorylation and/or dephosphorylation. PMID- 11853162 TI - Selective phosphodiesterase type 4 inhibitors reduce the prolonged survival of eosinophils stimulated by granulocyte-macrophage colony-stimulating factor. AB - It is well known that bronchial asthma is defined as chronic eosinophilic inflammation of the respiratory tract and that as one of the various types of inflammatory cells, eosinophils induce the airway inflammation of chronic asthma. Granulocyte-macrophage colony-stimulating factor (GM-CSF) has been shown to play an important role in the prolongation of the survival of eosinophils. We investigated the inhibitory effect of the selective phosphodiesterase (PDE) 4 inhibitors, 3,4-dipropyl-4,5,7,8-tetrahydro-3H-imidazo[1,2-i]purin-5-one (XT-611) and rolipram, and the nonselective PDE inhibitor theophylline, against GM-CSF induced prolongation of the survival of eosinophils isolated from patients with bronchial asthma. Eosinophils (10(6) cells/ml) were incubated in the presence of GM-CSF together with or without theophylline, rolipram or XT-611 at 37 degrees C, and the viable cells were assessed up to 4d using Trypan blue dye exclusion. The presence of theophylline (10(-4) M), rolipram (10(-4)-10(-5) M) or XT-611 (10(-4) 10(-5) M) significantly reduced the GM-CSF (10 pg/ml)-induced prolongation of viability of eosinophils. These findings suggest that selective PDE 4 inhibitors, including XT-611, may effectively reduce the activities of inflammatory cells in the airway of bronchial asthma patients. PMID- 11853163 TI - Antidiabetic action of low molecular weight chitosan in genetically obese diabetic KK-Ay mice. AB - Recently, we reported that low molecular weight (LMW) chitosan (chitosan lactate, average MW: 20,000) prevents the progression of low dose (100 mg/kg, i.p.) streptozotocin-induced slowly progressive diabetes mellitus in male ICR mice. The present study was designed to clarify the effects of LMW chitosan on hyperglycemia, hyperinsulinemia and hypertriglyceridemia in genetically obese diabetic male KK-Ay mice. LMW chitosan (0.05%, 0.2% or 0.8% water solution) was given daily as drinking water to male KK-Ay mice for 11 weeks, from 5 weeks of age. The non-fasting serum glucose levels of control mice continued to increase slowly throughout the experimental period. LMW chitosan lowered the serum glucose levels in a dose-dependent manner. In these diabetic mice, hyperinsulinemia and hypertriglyceridemia were observed, and LMW chitosan was dose-dependently effective in improving both serum biochemical parameters. LMW chitosan at three doses improved overdrinking and polyuria observed in these diabetic mice. It is concluded from these results that LMW chitosan may be useful for the treatment of obesity-related type 2 diabetes mellitus. PMID- 11853164 TI - Apoptosis induced by dioscin in Hela cells. AB - Dioscin, a saponin extracted from the root of Polygonatum Zanlanscianense Pamp, markedly inhibited proliferation of Hela cells. The results indicated that Hela cells underwent apoptosis in dose- and time-dependent manners when treated with Dioscin. Caspase-3, -8 and -9 activities were also detected. The low enzymatic activity of caspase-8 and high activity of caspase-9 showed that the mitochondrial pathway was activated in apoptosis. The reduced expression of the survival protein Bcl-2 also confirmed this result. These studies may be significant in finding a new drug to treat human cervical cancer. PMID- 11853165 TI - Safety of dietary supplements: chronotropic and inotropic effects on isolated rat atria. AB - We investigated the effects of dietary supplements on atria isolated from male Wistar rats. The examined supplements, which are increasingly used in Japan, those were Ginkgo biloba extract (GBE), catechins, isoflavones, sodium iron chlorophyllin and sodium copper chlorophyllin. GBE at 100-1000 microg/ml significantly increased the beat rate and the contractile force. Catechins at 1 100 microg/ml significantly potentiated the contractile force but did not effect the beat rates. However, isoflavones, sodium iron and sodium copper chlorophyllins did not change the contractile force or the beat rates. To identify the active ingredient of GBE, ginkgolide B, quercetin and amentoflavone on the atria were tested. Ginkgolide B weakened the contractile force. Quercetin potentiated the contractile force at only 30 microg/ml. Amentoflavone significantly increased the beat rate. From these findings, amentoflavone and quercetin were considered to be the principal ingredients of GBE producing the positive chronotropic and inotropic actions, respectively. In the case of catechins, (-)-epigallocatechin gallate (EGCg), one of the principal ingredients, produced inotropic actions. These findings suggest that there are some dietary supplements which affect cardiac function, such GBE and catechins. PMID- 11853166 TI - Inhibition of the antibody production by acetaminophen independent of liver injury in mice. AB - The causal relationship between the inhibition of antibody production and liver injury induced by single doses of acetaminophen (APAP) was investigated in mice. The liver injury and antibody production were evaluated using the serum transaminase activity and the number of antibody forming cells against sheep red blood cells (SRBC), respectively. The relevance of APAP hepatotoxicity with inhibiting antibody production was elucidated in fasted and fed mice treated with a single oral administration of APAP. In fasted mice, the oral administration of APAP produced serious liver injury, while it was not the case in the fed mice. As the antibody production was measured under these conditions, APAP significantly depressed the antibody production in fed mice as well as in fasted mice. The rate of B220 positive cells in the splenocytes was significantly decreased by APAP administration in both the fasted and fed mice. Splenocytes proliferative responses following mitogenic stimulation with concanavalin A or lipopolysaccharide were inhibited by APAP. Moreover, APAP added directly to the splenocyte culture also inhibited the in vitro antibody-producing response to SRBC. These findings indicate that the APAP-induced depression of antibody production may not be a secondary response to APAP-hepatitis, but may be a primary response to APAP. PMID- 11853167 TI - Testicular toxicity of rinbacin in rats. AB - Rinbacin is a local Nigerian herbal remedy. The effects of rinbacin on testicular histology were studied in prepubertal rats. Sexually immature male rats, divided into seven per group, were given rinbacin in drinking waters at 0, 26.25 g/l, or 52.50 g/l for 13 weeks, after which the animals were killed and testes excised, weighed, and processed for histologic study. The epididymal sperm number (ESN) was determined. There were no significant effects of either the low or high doses of rinbacin on fluid intake, body weight, testicular weight, and testis-body weight ratio. There was, however, a significant (p<0.05) decrease in the ESN of animals at both doses of rinbacin. Histologic examination of the testes indicated that the high dose of rinbacin induced significant degenerative changes, while the low dose had only a mild effect on testicular histology. Rinbacin decreases the ESN and causes degenerative lesions, especially at the high dose, in prepubertal rats. PMID- 11853168 TI - Phthalate esters detected in various water samples and biodegradation of the phthalates by microbes isolated from river water. AB - Phthalate esters (PEs), especially di-n-butyl phthalate (DBP) and di-(2 ethylhexyl) phthalate (DEHP) were detected in various water samples such as river water, well water and tap water. On degradation tests of PEs, Tempaku River water degraded almost 100% of diethyl phthalate (DEP), di-isobutyl phthalate and DBP, and approximately 70% of DEHP. All eight isolates from Tempaku River water (R1 R7, D1) did not degrade dimethyl phthalate (DMP), but showed biodegrading ability for the other PEs. The DBP-degrading ability was particularly high for the isolates R1-R3 and D1 of Acinetobacter iwoffii. Crude enzyme solutions prepared from bacterial cells of these isolates showed a higher degrading activity for DEHP compared with that for microbially-degradable DBP. Particularly high DEHP degrading activity was found for crude enzyme solutions of the isolate D1. As metabolites from the river water and bacterial isolates, DMP and an unknown diester were produced from DEP. DMP, DEP, monomethyl phthalate, monobutyl phthalate (MBP) and an unknown diester were produced from DBP. DBP, DEP, DMP and an unknown diester were produced from DEHP. As metabolites by the crude enzyme solutions, DMP, MBP and an unknown diester derivative were produced from DBP. DBP, mono-(2-ethylhexyl) phthalate and an unknown diester derivative were produced from DEHP. Diesters with shortened alkyl carbon chains were also found as metabolites by the isolates and their crude enzyme solutions. The results suggest that the alkyl chains in the diesters are also decomposed in addition to monoester formation from DBP or DEHP at the first step reported for animals and some types of bacteria. PMID- 11853169 TI - Synthesis and pharmacological activities of 2-(3'-substituted-2' hydroxypropylamino)pyridines. AB - In the present study, a series of 2-(3'-substituted-2' hydroxypropylamino)pyridines were synthesized and characterized by IR, 1H-NMR and elemental analysis. The compounds were investigated for anticonvulsant (150, 300 mg/kg) and cardiac activity. 2-(3'-Diethylamino-2'-hydroxypropylamino)pyridine 3 was found to exhibit the highest anticonvulsant activity. 2-(3'-Dimethylamino-2' hydroxypropylamino)pyridine 2 and 2-[3'-(4''-nitrophenylamino)-2' hydroxypropylamino]pyridine 6 were found to exhibit negative ionotropic activity. 2-(3'-Dimethylamino-2'-hydroxypropylamino)pyridine 2, 2-[3'-(4'' nitrophenylamino)-2'-hydroxypropylamino]pyridine 6 and 2-(3'-piperidino-2' hydroxypropylamino)pyridine 8 were found to antagonize exhibit beta-adrenergic activity. PMID- 11853170 TI - Pharmacognostical studies of cistanchis herba (III) phylogenetic relationship of the cistanche plants based on plastid rps2 gene and rpl16-rpl14 intergenic spacer sequences. AB - The phylogenetic relationship of Cistanche deserticola, C. salsa and C. tubulosa was analyzed by comparing the nucleotide sequences of the plastid rps2 gene and the intergenic spacer region between rpl16 and rpl14. By comparison of sequence data, the Cistanche samples were distinguishable from each other. The results were consistent with their anatomical and chemical characteristics. Intraspecific variations were found in C. salsa and C. tubulosa among the geographical populations. The NJ tree reconstructed based on the sequence data revealed that C. deserticola and C. salsa from China were closely related to each other, and C. tubulosa was placed as an outgroup of them. PMID- 11853172 TI - Inhibitory mechanism of an extract of Althaea officinalis L. on endothelin-1 induced melanocyte activation. AB - It is known that expression of endothelin-1 (ET-1) increases in the epidermis after UVB irradiation, and that this plays an important role during the induction of pigmentation both as a mitogen and as a melanogen for normal human melanocytes (NHMC). When ET-1 acts on NHMC via the endothelin B receptor (ET(B)R) on their cell surface, mobilization of intracellular calcium is induced, which is followed by activation of Raf-1 located upstream of mitogen activated protein kinase (MAPK). We have continued the search for new agent which inhibit this calcium mobilization and we have found that an extract of Althaea officinalis L. has such an action. In this study, we investigated the precise inhibitory mechanism of this botanical extract on the ET-1-induced activation of melanocytes. Treatment of NHMC with this extract abrogated the stimulatory effect of ET-1 on proliferation and also on activation of MAPK in the intracellular signal transduction pathway, but did not affect the binding of ET-1 to the ET(B)R or the production of Inositol 1,4,5-Trisphosphate (IP3). Further, when this extract was used to treat normal human keratinocytes (NHKC), secretion of ET-1 by those cells was reduced. Taken together, these findings indicate that an extract of A. officinalis inhibits both the secretion of ET-1 from NHKC and the action of ET-1 on NHMC mainly by suppressing the ET-1-induced calcium mobilization without the modification of IP3 production, which in turn suggests that this extract is a useful ingredient for a whitening agent. PMID- 11853171 TI - Inhibition of cell cycle progression through specific phase by pancratistatin derivatives. AB - Pancratistatin derivatives, 1-O-(3-hydroxybutyryl)pancratistatin (HBP) and 1-O-(3 O-beta-D-glucopyranosylbutyryl)pancratistatin (GBP), showed strong cytostatic activity against rat embryo fibroblast 3Y1 at concentrations less than 1 microM. When the effect on cell cycle progression was examined in 3Y1 fibroblasts arrested at G0/G1 phase by serum deprivation, HBP, GBP, and pancratistatin inhibited the progression of 3Y1 fibroblasts from G0/G1 to S phase. In addition, when the effect on cell cycle progression was studied in 3Y1 fibroblasts synchronized at late G1/early S phases by treating with hydroxyurea, HBP blocked further progression through S phase, while GBP and pancratistatin did not affect the progression, but retarded it. On the other hand, when the effect of HBP and GBP on the progression was evaluated in promyelocytic leukemia HL-60RG cells synchronized at G0/G1 phase, the cells did not progress into S phase and accumulated in sub G0/G1 phase, which indicated apoptotic cells. These findings suggest that of Amaryllidaceae alkaloids, HBP blocks the progression of cell cycle at least at G0/G1 and S phases and GBP does at least at G0/G1 phase, resulting in apoptosis induction in tumor cells. PMID- 11853173 TI - Inhibition of human hepatic cytochrome P450s and steroidogenic CYP17 by nonylphenol. AB - Effect of nonylphenol on aminopyrine N-demethylase activity, a typical drug metabolizing enzyme activity, by ten kinds of human hepatic cytochrome P450s (CYP) and on progesterone 17alpha-hydroxylase activity by steroidogenic CYP17 was investigated. When determined at 2 mM substrate concentration, nonylphenol (1 mM) most efficiently inhibited aminopyrine N-demethylation by CYP2C9 and CYP2C19, by 61% and 59%, respectively, followed by CYP2D6, CYP1A2, CYP2C18 and CYP2C8 (46 51%), whereas inhibition of the activities by other CYPs was less than 27%. Additionally, nonylphenol competitively inhibited diclofenac 4'-hydroxylation by CYP2C9 and S-mephenytoin 4'-hydroxylation by CYP2C19 with Ki values of 5.3 and 37 microM, respectively. Furthermore, nonylphenol exhibited a competitive inhibition of progesterone 17alpha-hydroxylase activity by CYP17 with Ki value of 62 microM. These results suggest that nonylphenol inhibits human hepatic CYPs, especially CYP2C9 and CYP2C19, and steroidogenic CYP17 activities. PMID- 11853174 TI - Differential mRNA expression of inflammatory cytokines in cultured human fetal membrane cells responding to influenza virus infection. AB - We examined the expression of mRNAs for inflammatory cytokines and Fas in cultured human fetal membrane cells responding to influenza virus (IV) infection using the reverse transcriptase-polymerase chain reaction (RT-PCR). Primary cultured chorion and amnion cells prepared from human fetal membranes were infected with IV. Chorion cells expressed significant amounts of interleukin (IL) 1beta, IL-6, tumor necrosis factor (TNF)-alpha, interferon (IFN)-beta, IFN-gamma and granulocyte macrophage colony-stimulating factor (GM-CSF) mRNAs and small amounts of Fas mRNA in response to IV infection. Amnion cells expressed TNF-alpha and IFN-beta mRNAs in response to IV infection, while expression of the other mRNAs was not altered. We also examined whether or not TNF-alpha, IFN-beta, IFN gamma and Fas participated in IV infection-induced apoptotic DNA fragmentation in chorion cells. Neutralizing antibodies against them did not inhibit DNA fragmentation. These results suggested that chorion cells expressed significant amounts of mRNAs for inflammatory cytokines in response to IV infection, and that, in contrast, mRNA expression was quiescent in amnion cells. Moreover, TNF alpha, IFN-beta, IFN-gamma and Fas do not appear to be directly involved in the apoptosis induction of IV-infected chorion cells. The results indicated that chorion cells may play a role in defense against IV through an antiviral immune response and apoptosis to eliminate own cells and viral pathogens in infected organs, whereas amnion cells do not play such a role. PMID- 11853175 TI - Studies on cancer chemoprevention by traditional folk medicines XXIV. Inhibitory effect of a coumarin derivative, 7-isopentenyloxycoumarin, against tumor promotion. AB - 7-Isopentenyloxycoumarin (1) was isolated from Heracleum lanatum MICHX. (Umbelliferae). Compound 1 inhibited phospholipid metabolism and Epstein-Barr virus activation caused by a potent tumor promoter. In an in vivo experiment, topical application of 1 suppressed skin-tumor-formation induced by 12-O tetradecanoylphorbol-13-acetate (TPA) in 7,12-dimethylbenz[a]anthracene (DMBA) initiated mice. And it also suppressed ornithine decarboxylase activity stimulated by TPA on mouse skin. These results indicated that 7 isopentenyloxycoumarin is one of the effective compounds from natural resources for treating skin tumor formation. PMID- 11853176 TI - Studies on cancer chemoprevention by traditional folk medicines XXV. Inhibitory effect of isoliquiritigenin on azoxymethane-induced murine colon aberrant crypt focus formation and carcinogenesis. AB - Isoliquiritigenin is a natural pigment with the simple chalcone structure, 4,2',4'-trihydroxychalcone. The effect of this compound on azoxymethane (AOM) induced colonic aberrant crypt focus and tumor formation in ddY mice was examined. Administration of 15 ppm of isoliquiritigenin in drinking water, significantly suppressed AOM-induced aberrant crypt focus formation (p<0.01), with an inhibitory ratio of 37.3%. Isoliquiritigenin also inhibited AOM-induced colon carcinogenesis by administration in a mixed diet. The average number of tumors was 14.6+/-8.9 items in the control group and were 7.3+/-7.3, 3.9+/-5.6, 4.7+/-6.5 items in the 10, 100 and 250 ppm in the isoliquiritigenin treated groups, respectively. In histopathological studies, the tumors were identified as adenoma and adenocarcinoma, however, significant differences were not observed between the control group and isoliquiritigenin treated groups. These results indicated that isoliquiritigenin might be a potential chemopreventive agent against colon cancer. PMID- 11853177 TI - The effects of several vasopressin receptor antagonists on normal intraocular pressure and the intraocular distribution of vasopressin receptor subtypes. AB - The aim of the present paper is to study the relation between vasopressin antagonism and the regulation of intraocular pressure (IOP). From the studies on the effect of several vasopressin receptor antagonists, VP-343, OPC-21268, YM087, OPC-31260 and SR121463, on normal IOP and the effect of VP-343 on pupil diameter in rabbit, it was shown that some vasopressin antagonists decreased normal IOP and VP-343 had no influence on pupil diameter. A vasopressin receptor mapping study in normal cynomolgus monkey eye revealed a high density binding site for a [H3]vasopressin V1 antagonist in the region of iris. These findings suggest that a vasopressin antagonist should decrease normal IOP without miosis and that vasopressin V1 receptors are present in iris. PMID- 11853178 TI - Antiallergic effect of flavonoid glycosides obtained from Mentha piperita L. AB - Six flavonoid glycosides, eriocitrin (1), narirutin (2), hesperidin (3), luteolin 7-O-rutinoside (4), isorhoifolin (5), diosmin (6), rosmarinic acid (7) and 5,7 dihydroxycromone-7-O-rutinoside (8), were isolated from the aerial part of Mentha piperita L. Among these compounds, compound 4 showed a potent inhibitory effect on histamine release induced by compound 48/80 and antigen-antibody reaction. This compound was more effective than luteolin and luteolin-7-O-glucoside in inhibiting histamine release from rat peritoneal mast cells. Compound 4 also caused a dose-related inhibition of the antigen-induced nasal response and significant effects were observed at doses of 100 and 300 mg/kg. These results indicate that compound 4 may be clinically useful in alleviating the nasal symptoms of allergic rhinitis. PMID- 11853179 TI - Antipruritic effect of Cnidii Monnieri Fructus (fruits of Cnidium monnieri CUSSON). AB - Antipruritic effects of 70% ethanol extract (CM-ext) of Cnidii Monnieri Fructus (dried fruits of Cnidium monnieri CUSSON, Umberifferae) were investigated. In mice, an oral administration of CM-ext (200 and 500 mg/kg) inhibited compound 48/80-induced scratching behavior without influence on spontaneous locomotion. Isopimpinellin (3) and osthol (1), coumarin derivatives isolated from CM-ext, showed an inhibitory effect on compound 48/80-induced scratching behavior. PMID- 11853180 TI - Comparative absorption of 5-aminosalicylic acid (5-ASA) after administration of a 5-ASA enema and salazosulfapyridine (SASP) after an SASP suppository in Japanese volunteers. AB - Salazosulfapyridine (SASP) is widely used orally and rectally in the treatment of ulcerative colitis. SASP is mainly metabolized by hydrolysis and the main active metabolite, 5-aminosalicylic acid (5-ASA), has an antiinflammatory effect. In the present study, we prepared suppositories containing 6.5 mmol of SASP and an enema containing 6.5 mmol of 5-ASA. We measured the concentrations of SASP and its various metabolites, 5-ASA, sulfapyridine (SP), acetylated metabolite of SP (Ac SP), and N-acetyl-5-ASA (Ac-5-ASA), in the serum and urine after a single administration of each preparation to healthy male volunteers. When the SASP suppository was administered, the maximum concentration (Cmax) of SASP and Ac-5 ASA was 2.5+/-0.4 and 0.5+/-0.2 microM and the time to Cmax (Tmax) was 5 and 12 h, respectively. The Cmax value of SP, which causes side effects, was one-half of that of the parent compound. No 5-ASA in the serum was observed. When the 5-ASA enema was administered, Cmax and Tmax values of 5-ASA and Ac-5-ASA were 5.8+/-2.0 and 13.3+/-3.6 microM and 1 and 7 h, respectively. The area under the serum concentration-time curve (AUC) of SASP was 27.4+/-4.8 microM x h, a finding similar to that of 5-ASA after the administration of the 5-ASA enema (29.4+/-11.1 microM x h). The percentage of urinary recovery of SASP 24 h after administration of the SASP suppository was approximately 0.2%. These results indicate that SASP administered rectally is almost completely hydrolyzed in the colon and that 5-ASA is partially absorbed from the small intestine in unchanged form. On the other hand, approximately 0.3% of 5-ASA was recovered in the urine in unchanged form after the administration of the 5-ASA enema, whereas the urinary recovery of Ac-5 ASA was more than 10%. The present findings suggest that 5-ASA has favorable absorptive properties and can be expected to have systemic action after rectal administration of a 5-ASA enema. PMID- 11853181 TI - Effect of chondroitin sulfate on the biodegradation and drug release of chitosan gel beads in subcutaneous air pouches of mice. AB - Chitosan (CS) gel beads were prepared in 10% amino acid solution (pH 9) and modified by forming an electrostatic complex between the amino group of CS and the carboxyl group of chondroitin sulfate (Cho). Modification of the CS gel matrix by Cho inhibited the in vitro release of prednisolone (PS) from the gel beads. CS gel beads modified by Cho (CS-Cho) were implanted into air pouches (AP) prepared subcutaneously on the dorsal surfaces of mice. No inflammatory response was observed. The in vivo release of PS from CS-Cho gel beads and their biodegradation in the AP was slower than beads without Cho treatment. After 28 days of implantation, CS-Cho gel beads (deacetylation of CS: 90%) were still detectable, although they had become softer and smaller. Modification of the CS gel matrix by Cho controls the biodegradation of the beads and the release of the drug. This effect makes these beads a promising biocompatible and biodegradable vehicle for sustained drug delivery. PMID- 11853182 TI - Preliminary screening of the inhibitory effect of food extracts on activation of the aryl hydrocarbon receptor induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin. AB - A preliminary screening for the inhibitory effects on the activation of the aryl hydrocarbon receptor (AhR) by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) by applying AhR-based bioassays for dioxins, the Ah-Immunoassay and CALUX assay, was attempted. Thirty-nine food extracts including vegetables, fruits, herbs, and teas were initially screened in vitro. We first examined the application of both bioassay methods using green tea extracts and (-)-epigallocatechin gallate, reported antagonists of the AhR, since the results could reveal an inhibitory effect versus the control in both assays. Food extracts were then tested. Among the herbs, extracts of sage, among the vegetables, green leafy ones such as spinach, and among the fruit, citrus showed inhibitory effects on AhR activation by TCDD, although some tested samples did not show parallel behavior in both assays. Sage had a remarkable inhibitory effect (79% in the CALUX assay and 83% in the Ah-Immunoassay compared with control) and its effects were dose dependent. The results suggest that these assays might be applicable to the preliminary screening of antagonist activity against the AhR. Moreover, based on these results, the potential benefit of factors that function as dietary ligands of the AhR and are present in several foodstuffs is indicated. PMID- 11853183 TI - Current state of screening for Down's syndrome. AB - Down's syndrome has an incidence of about 1.3 per 1000 births. Antenatal testing for this disorder on the basis of maternal age has been common practice (increasing age is associated with increasing risk of having an affected pregnancy). Since the late 1980s it has been possible to calculate a pregnancy specific risk using maternal age and serum marker levels, but the absence of a national screening policy in the UK has resulted in a diversity of screening practice. Screening may take place in the first or second trimester of pregnancy, using maternal age in combination with up to six serum markers, with or without ultrasound markers. Additionally, some units offer the screen only to women in specific age groups. Risk and marker levels are affected by a number of factors for which adjustments in the calculation may be made. Laboratories offering screening must also establish robust monitoring and quality management systems to ensure continued confidence in the reported result. Current diagnostic techniques require the use of invasive procedures which pose a risk to the fetus and the mother. Future screening may combine new technologies with the identification of fetal cells in maternal blood to reduce or abolish the need for these procedures. PMID- 11853184 TI - HDL-cholesterol and cardiac disease: which table to use? AB - Interventions to modify cardiovascular risk factors have the greatest benefit in individuals at greatest risk of a cardiovascular event, not necessarily those with the most extreme levels of any particular risk factor. Patients with vascular disease are readily identified, but risk calculation in those without disease provides a way of integrating several risk factors to identify individuals at greatest risk. Equations have been derived from different epidemiological studies (Framingham, PROCAM, etc.) to estimate the risk of different clinical end-points [coronary heart disease (CHD) or cardiovascular disease (CVD)]. Some algorithms have been adapted to produce risk charts' or 'tables'; others have been modified to incorporate additional risk factors. Current UK guidelines all endorse calculation of CHD risk using the Framingham algorithm. This predicts risk well for some North European populations but less reliably in low-risk populations. It does not incorporate some risk factors, including serum triglycerides, family history, C-reactive protein, or homocysteine. Risk calculation should not be used in those with evidence of vascular disease, genetic hyperlipidaemia or a strong family history of CHD, and should be interpreted with caution in non-Caucasians. Measurement of high-density lipoprotein (HDL)-cholesterol is essential for accurate risk calculation. This has significant cost and workload implications for the laboratory. The way a laboratory reports results should be designed in such a way as to facilitate simple and accurate risk calculation. PMID- 11853185 TI - Gamma-glutamyltransferase: value of its measurement in paediatrics. AB - Gamma-glutamyltransferase (GGT) is a microsomal enzyme that is widely distributed in human tissues involved in secretory and absorptive processes, particularly the bile canaliculi. Serum GGT is elevated in liver diseases affecting the biliary system, such as extrahepatic biliary atresia, sclerosing cholangitis and progressive familial intrahepatic cholestasis (PFIC) type 3. Conversely, two other subtypes of PFIC have normal or low serum GGT activity, discordant with the degree of cholestasis. GGT is also useful in detecting cholestasis associated with parenteral nutrition and assessing the efficacy of ursodeoxycholic acid in its treatment. GGT may also help screening for biliary complications on patients that have undergone orthotopic liver transplantation. The reference range for GGT is age dependent. In normal full-term neonates the activity at birth is approximately six to seven times the upper limit of the adult reference range. The activity then declines, reaching adult levels by the age of 5-7 months. PMID- 11853186 TI - Audit of investigation of subclinical hypothyroidism in Scottish laboratories. AB - BACKGROUND: An audit of thyroid function testing in the investigation of subclinical hypothyroidism in Scotland was performed. METHODS: The audit looked at the tests used and the strategies employed by 18 out of a total of 25 Scottish laboratories and found significant differences between them. It also looked at the comments and advice given by 59 clinical scientists and chemical pathologists in the interpretation of results from patients with subclinical hypothyroidism. These comments were compared with standards derived from guidelines in current literature. RESULTS: The advice given was generally accurate and helpful but the audit showed a marked variation in interpretative comments. CONCLUSIONS: This variation may reflect the lack of emphasis that has been placed on standardizing the interpretive function of laboratories and underlines the need for liaison both within and between laboratories to achieve better agreement on the interpretation of laboratory tests. The audit also showed that the importance of thyroid peroxidase antibody estimation in subclinical hypothyroidism needs to be emphasized. PMID- 11853187 TI - Novel nephelometric assay for measurement of complement 3d. AB - BACKGROUND: This report describes a nephelometric assay for measuring complement 3d (C3d). METHODS: C3d was separated from C3 and the other C3 split products by polyethylene glycol precipitation. The supernatant containing C3d was then measured by rate nephelometry in a Beckman Immage rate nephelometer using a specific antibody. Calibration was performed with dilutions of a standard, which was obtained by incubating a serum pool at 37 degrees C for 7 days. This incubation step allowed the activation and breakdown of C3 into C3d. RESULTS: The assay was linear across the normal and pathological range. Precision studies showed within-run and between-run coefficients of variation of < 5% and < 6%, respectively. The nephelometric results are comparable with those obtained by radial immunodiffusion. Neither haemoglobin nor lipids interfere with the assay. PMID- 11853189 TI - Identification and determination of salicylic acid and salicyluric acid in urine of people not taking salicylate drugs. AB - BACKGROUND: Salicylic acid (SA) is present in the serum of people who have not taken salicylate drugs. Now we have examined the urine of these subjects and found that it contains SA and salicyluric acid (SU). We have established the identities of these phenolic acids and determined their concentrations. METHODS AND RESULTS: The acidic hydrophobic compounds of urine were separated using high performance liquid chromatography (HPLC) and were detected and quantified electrochemically. Two approaches were used to establish the identity of SA and SU. First, the retention times (Rt) of the substances extracted and those of SA and SU were compared under two sets of chromatographic conditions; the Rt of the compounds suspected to be SA and SU and those of the authentic substances were very similar under both sets of conditions. Second, the unknown substances, isolated by HPLC, were treated with acetyl chloride in methanol and compared with the methyl esters of SA and SU by using gas chromatography-mass spectrometry; the unknown compounds after esterification had very similar mass spectra and gas chromatographic R, to those of methyl salicylate and methyl salicylurate. The median (n = 10) urinary concentration of SA was 0.56 micromol/L (range 0.07-0.89 micromol/L) and that of SU was 3.20 micromol/L (range 1.32-6.54 micromol/L). SA and its major urinary metabolite, SU, were found in the urine of all of the 10 people examined. PMID- 11853188 TI - Molecular analysis of a thai beta-thalassaemia heterozygote with normal haemoglobin A2 level: implication for population screening. AB - BACKGROUND: This study aimed to characterize the molecular basis of a beta thalassaemia heterozygote who had a normal haemoglobin A2 level. METHODS: Using haematological and molecular analyses, the alpha- and beta-globin genotypes in a Thai woman with normal haemoglobin A2 level, who was identified in a thalassaemia and haemoglobinopathy screening programme, were examined. RESULTS: The patient was positive for a 0.36% one-tube osmotic fragility test but negative for the dichlorophenolindophenol dye test for haemoglobin E. Haemoglobin A and A2 were observed on cellulose acetate electrophoresis with the haemoglobin A2 of 2.0%, compatible with an alpha-thalassaemia 1 carrier. Polymerase chain reaction analysis failed to detect alpha-thalassaemia 1 (South East Asian deletion). Beta globin gene analysis detected a severe betao-thalassaemia allele with the 4 bp ( CTTT) deletion at codons 41/42. Further analysis of the alpha-globin gene identified the homozygosity of the 3.7 kb deletion alpha-thalassaemia 2. CONCLUSIONS: The patient was found to be an unusual case: a betao-thalassaemia carrier with a low haemoglobin A2 concentration, in combination with a homozygosity for the 3.7 kb alpha-thalassaemia 2 deletion. The approach used to characterize the patient and the result obtained in this report will prove useful for population screening of thalassaemia in regions where both alpha- and beta thalassaemias are common. PMID- 11853190 TI - Apolipoprotein A-I concentration in tears in diabetic retinopathy. AB - BACKGROUND: Apolipoprotein concentrations in reflex tears from healthy and noninsulin-dependent diabetes mellitus (NIDDM) subjects were measured and correlated with the stage of diabetic retinopathy. METHODS AND RESULTS: The apolipoprotein A-I (apo A-I) concentrations in the tears from NIDDM patients with retinopathy were significantly higher than those from patients with no or negligible retinopathy (P<0.05), and apo A-I was not detected in healthy subjects by Western blotting. No reactive band was detected on apo A-I by Western blotting for nitrotyrosine, a marker for peroxynitrite oxidation of the tear proteins. Apo A-I concentration in tears was significantly correlated with the stage of retinopathy (r= 0.598, n = 59, P < 0.001). No apo A-I gene expression was detected in the conjunctiva by reverse transcription polymerase chain reaction. CONCLUSIONS: We conclude that there is increased secretion of native apo A-I from the main lacrimal gland in patients with advanced diabetic retinopathy. PMID- 11853191 TI - Artefactual reduction in plasma total carbon dioxide concentration in children. AB - METHODS: The effect on plasma total carbon dioxide (ctCO2) of collecting a small volume of blood into a large tube was investigated. In a 7-month-old infant presenting with diarrhoea the initial ctCO2 was low when 0.3 mL of sample was received in a 4-mL tube. To investigate this further blood samples were obtained from three volunteers on five occasions and various volumes were placed in a 4 mL container and analysed after varying intervals. RESULTS: When the sample volume was less than 2 mL there was a significant decrease in ctCO2. A 42% decrease was seen in the tube containing 0.3 mL after 4 h. CONCLUSIONS: We recommend that when a small volume of sample is collected it should be placed in a smaller tube with minimal air space. PMID- 11853192 TI - Spot urine analysis: acidification does not increase calcium recovery. AB - INTRODUCTION: We investigated the effects of acidification and storage on calcium recovery in spot urine samples. METHODS: In a retrospective study, 43 spot urine samples from patients with metabolic bone disease, which had been previously analysed fresh for calcium and creatinine, were acidified and incubated for 2 h at room temperature and re-analysed. In a prospective study, fresh spot urine samples from 15 volunteers were split into two aliquots--one left unacidified and the other acidified with 6 mol/L HCl to pH < 2. Both aliquots were analysed immediately and after storage at 4 degrees C for 24 h. RESULTS: In both studies, the calcium creatinine ratios obtained on acidified and non-acidified spot urine samples were not different, both when measured immediately as well as after storage at 4 degrees C for 24 h. CONCLUSIONS: The acidification of spot urine samples would thus appear to be unnecessary for calcium/creatinine measurements in metabolic bone disease if they were undertaken within at least 24 h. PMID- 11853193 TI - Effect of low protein concentration on serum sodium measurement: pseudohypernatraemia and pseudonormonatraemia! AB - BACKGROUND: The effect of high concentrations of protein or lipid on the measurement of plasma sodium by indirect ion selective electrode (ISE) causing pseudohyponatraemia and pseudonormonatraemia is well described. The effect of a low total protein concentration, however, has not been described. METHODS: In order to examine this, over a 2-week period the total protein concentration was measured on all samples received for urea and electrolyte measurement. All samples with a low (<50 g/L) or a high (> 80 g/L) total protein concentration had sodium measured by both direct and indirect ISE. RESULTS: There were approximately equal numbers of samples with a protein concentration less than 50 g/L (1.3%) as samples with a protein concentration greater than 80 g/L (1.3%). The frequency of erroneous sodium results owing to the use of an indirect ISE was less in hypoproteinaemic (2%) than in hyperproteinaemic (20%) samples. PMID- 11853194 TI - Effect of breast-feeding on concentration of nitric oxide in breast milk. AB - BACKGROUND: The effect of breast-feeding on the concentration of nitric oxide (NO) in milk was investigated in lactating mothers. METHODS: Breast milk specimens were obtained from 45 lactating women after normal delivery on postpartum days 1, 2, 3, 4 and 5. The subjects were divided into three groups: group 1, breast-feeding only; group 2, mixed feeding (breast milk by manual expression and additional formula feeds); and group 3, formula feeding only. Milk NO concentration was measured as the sum of nitrite and nitrate using the Griess reaction. RESULTS: The highest mean milk NO concentration was on day 4 in all groups, but the peak value in group 3 was significantly lower (P<0.0001) than in other groups. The milk NO concentrations of group 1 were significantly greater than those of groups 2 and 3. CONCLUSIONS: We conclude that breast-feeding increases the concentration of NO in milk. PMID- 11853195 TI - Correlation between serum magnesium and blood cyclosporine A concentrations in renal transplant recipients. AB - BACKGROUND: Low total serum magnesium concentration has been reported in renal transplant recipients on cyclosporine therapy, and this is a risk factor for hypertension and cardiac death. No study is yet available from the Indian subcontinent on the relationship between total serum magnesium and cyclosporine A concentration. METHODS: Blood cyclosporine A and total serum magnesium concentrations were estimated in 31 post renal transplant patients. Sixteen chronic renal failure patients who had not yet received a transplant served as controls. RESULTS: The mean total serum magnesium in post-transplant patients was 0.64+/-0.11 mmol/L (n = 31), which was significantly lower (P <0.001) than in the controls (1.0+/-0.16 mmol/L). An inverse correlation was found between total serum magnesium and blood cyclosporine A concentration (r = -0.532). Systolic and diastolic blood pressures were also higher in the cyclosporine-treated patients. CONCLUSIONS: We conclude that serum magnesium should be monitored regularly in patients receiving cyclosporine A. PMID- 11853196 TI - Quantitative determination of plasma phenylalanine and tyrosine by electrospray ionization tandem mass spectrometry. AB - BACKGROUND: Phenylketonuria patients on protein-restricted diets require monitoring of their blood phenylalanine concentrations. Small sample volumes are essential for neonates and infants, with the need for results to be generated in a timely manner by robust methods. We present a tandem mass spectrometry assay for phenylalanine (and tyrosine) which fulfils these criteria. METHODS: Plasma samples were deproteinized using a methanolic solution of phenylalanine-d5 and tyrosine-d4 that served as the internal standards for the assay. Following centrifugation each liquid phase was dried in a well of a microtitre plate and the amino acids converted to butyl esters using butanolic hydrogen chloride. After drying again and reconstituting in 80% acetonitrile in water, the samples were analysed using electrospray ionization tandem mass spectrometry. RESULTS: For both phenylalanine and tyrosine the limit of detection was 1 micromol/L and the carry-over less than 1%. The assay was linear to 3000 micromol/L in both amino acids (phenylalanine: r= 0.999; tyrosine: r= 0.992). Inter-sample precision varied with concentration but was 3.4-5.2% for phenylalanine and 4.3-7.6% for tyrosine. Between-batch precision was more variable, being 4.1-12.9% for phenylalanine and 8.4%-10.5% for tyrosine. CONCLUSIONS: Good agreement was found between the assay and results from existing ion-exchange chromatography and high performance liquid chromatographic assays in this department. Carry-over and precision were acceptable for monitoring work and the linearity allowed most specimens to be analysed without dilution. Where available, tandem mass spectrometry offers a convenient means of transferring the routine work for both neonatal screening and monitoring to one instrument. PMID- 11853197 TI - Anaphylactic reaction to intravenous sinistrin (Inutest). AB - Inulin clearance is the 'gold standard' for the determination of glomerular filtration rate. This is the first report of anaphylaxis following intravenous administration of inulin. PMID- 11853198 TI - Faecal fat: time to give it up. PMID- 11853199 TI - Standardization of glycated haemoglobin. PMID- 11853200 TI - A new method for determining patient payments for outpatient drugs. PMID- 11853201 TI - Expanded use of nurse practitioners. PMID- 11853202 TI - Evaluation of agreements between managed care organizations and providers of community-based mental illness and addiction disorder treatments. AB - OBJECTIVE: To analyze the scope, content, and nature of contracts between managed care organizations (MCOs) and community-based mental illness and addiction disorder treatment and prevention service providers, focusing on implications for managed care policy. STUDY DESIGN: Qualitative and quantitative document content analysis. METHODS: We reviewed and coded 107 provider contracts from 17 states. Data were aggregated on MCO and provider demographics, scope of services, contractual obligations, and financial reimbursement mechanisms. Results were compared with those from a similar study conducted in 1996 with a sample of 50 contracts to identify changes over time. RESULTS: The MCOs purchased relatively few services from providers, omitting many services integral to the proper management of mental illness and addiction disorders. Service duties were often ambiguously described, leading to potentially significant and unanticipated financial risk for providers. The MCOs exert strong control over treatment decision making. Capitation and other risk-based payment arrangements are increasingly common, although most continue to use fee-for-service reimbursement methods. Contracts are structured to remove provider bargaining power; they allow MCOs to unilaterally amend all provisions on notice and without negotiation and permit termination "at will." CONCLUSIONS: Managed care contracts favor the needs of the managed care industry and are constructed to (1) shift significant amounts of financial risk onto providers and (2) manage and restrain providers' choices over the use of benefits through close oversight, financial controls, and incentives. Because a signed contract is a precondition to access to patients and insurance revenues, health professionals must sign them and indicate a general inability to negotiate their terms. PMID- 11853203 TI - "Trial" prescriptions to reduce drug wastage: results from Canadian programs and a community demonstration project. AB - OBJECTIVE: To determine the acceptability of a program to avoid drug wastage through "trial" prescriptions. STUDY DESIGN: Cross-sectional survey, followed by a 9-month demonstration project. METHODS: Consultants to trial prescription programs operated by 2 public- and 3 private-sector drug plans in Canada were surveyed. All of the trial prescription programs were voluntary. The demonstration project involved 215 English-speaking adults who filled a "new" prescription for an angiotensin-converting enzyme inhibitor, beta-blocker, or calcium channel blocker (CCB) in 1 of 16 Peterborough, Ontario pharmacies. Patients received a 7-day supply of medication and a reminder card on which the dispensing pharmacist recorded the mutually agreed date and time the patient would be contacted to assess the results of the "trial." Patients who tolerated the medication received the balance of their original prescription. RESULTS: Most patients (86%) who were offered trials in the demonstration project accepted them, and most (82%) who accepted them found them helpful. The proportion of patients who received the balance varied by program, ranging from 47.1% to 86.6%. The dollar value of the wastage avoided through trial prescriptions varied by drug class. This was driven largely by differences in the unit cost of the medications, but also to a lesser extent by larger prescriptions for CCBs. CONCLUSIONS: Trial prescriptions were acceptable to patients and, if focused on specific medications, could reduce the direct cost of drug wastage. More work is needed to define the conditions under which trial prescription programs are feasible for pharmacists and drug plans. PMID- 11853204 TI - Comparison of 2 systems for clinical practice profiling in diabetic care: medical records versus claims and administrative data. AB - OBJECTIVES: To (1) describe the rate of glycosylated hemoglobin (HbA1c) testing and control in a primary care clinic at an academic medical center; (2) compare academic medical center and health maintenance organization (HMO) data for a subgroup of the same patients; and (3) discuss the challenges to accurate clinical practice profiling. STUDY DESIGN: Retrospective analysis of medical records and claims and administrative data. PATIENTS AND METHODS: We reviewed the medical records of 300 patients with diabetes mellitus from the Primary Care practice at an academic medical center (AMC). The HMO and AMC were in the southeastern United States. The study adhered to the National Committee for Quality Assurance's Health Plan Employer Data and Information Set 3.0 (1998) guidelines for collection of the numerator and denominator data. RESULTS: Overall, 275 (91.7%) of 300 patients underwent a documented HbA1c test (range, 77.8%-98.0% across the 5 Primary Care Associates clinic physicians). Of these 275 patients, 206 (74.9%; range, 52.4%-84.7%) were under "good control" (defined by the National Committee for Quality Assurance as an HbA1c level of < or = 9.5%). The mean (SD) HbA1c level was 8.45% (1.98%). Data from the HMO documented a rate of HbA1c testing of 36.3% (45/124) compared with 92.7% (115/124) based on case note review at the academic medical center. CONCLUSIONS: Current administrative and claims-based information systems have inherent weaknesses if used for performance measurement. Reliance solely on medical record review is time and cost prohibitive. To ensure complete reporting of mandated "quality measures" will necessitate the tracking of data across different healthcare systems. PMID- 11853205 TI - A comparison of ambulatory services for patients with managed care and fee-for service insurance. AB - OBJECTIVES: To determine whether family physicians provide different ambulatory care to patients with health insurance from managed care organization (MCO) versus fee-for-service (FFS) plans. STUDY DESIGN: Multimethod cross-sectional observational study. PATIENTS AND METHODS: A total of 4454 patients made office visits to 138 family physicians in northeastern Ohio. Direct observation with the Davis Observation Code and a structured checklist were used to assess the process of care. Patient satisfaction was measured with the Medical Outcomes Study 9-Item Visit Rating Form. RESULTS: Among 1588 patients with MCO insurance and 876 with FFS insurance, no differences were noted in the number of visits per year, length of visits, percentage of visits for well care, or the percentage of visits in which medicines were prescribed in analyses controlling for patient mix. Visits by patients with MCO insurance were more likely to involve referrals to another physician than visits by patients with FFS insurance. Patient satisfaction and time use during visits were comparable for the 2 groups. CONCLUSIONS: Managed care insurance appears to increase involvement of the primary care provider in the referral process. However, in a healthcare market with moderate managed care penetration in which the same physicians see patients with different types of insurance, the process of care is affected little by type of insurance coverage. PMID- 11853206 TI - Accomplishing the impossible: talking about body and soul and mind during a medical visit. AB - This study examines the practical side of communicating about patients' thoughts, feelings, and physical health. A study of 53 medical visits suggests that biopsychosocial communication need not be as difficult or as time consuming as many practitioners believe. Biopsychosocial medical visits can be conducted in roughly the same amount of time as biomedical visits with many important advantages. This report describes 6 communication techniques used by 1 doctor to encourage talk about lifeworld issues. The techniques include self-disclosing, expressing empathy, involving patients in decision-making, talking openly about patients' fears, asking open-ended questions, and listening attentively. The doctor studied was able to use these techniques without exceeding the average length of a biomedical visit. PMID- 11853207 TI - Beyond the organ donor card: the effect of knowledge, attitudes, and values on willingness to communicate about organ donation to family members. AB - Although numerous studies have examined many of the predictors of signing an organ donor card, including knowledge, attitudes, values, and demographic variables, very few have examined the factors associated with individuals' willingness to communicate about organ donation with family members. Because organ donation does not take place without the permission of a person's next-of kin, government agencies and organ procurement organizations have targeted communication with family members as a primary objective of organ donation campaigns. This study reports the results of a survey of a stratified random sample of adults at 2 local sites of a national employer. Results indicate that knowledge, attitude, and altruism are significantly related to 2 measures of willingness to communicate: past behavior (whether respondents had already discussed organ donation with family members) and a scale measuring willingness to communicate about organ donation in the future. Because the quality of discussions between the potential donor and his or her family will depend on how well the donor is able to address vital issues regarding donation, it is concluded that campaigns seeking to promote communication between family members about organ donation must simultaneously seek to increase knowledge, debunk myths, and bolster positive attitudes about donation. PMID- 11853208 TI - Predictors of exposure from an antimarijuana media campaign: outcome research assessing sensation seeking targeting. AB - Using data from a large-scale antimarijuana media campaign, this investigation examined the demographic and psychographic variables associated with exposure to public service announcements designed to target high sensation-seeking adolescents. The literature on sensation seeking indicates that adolescents high in this trait are at greater risk for substance abuse. Analyses assessed the predictive utility of various risk and protective factors, normative influences, demographic variables, and marijuana-related attitudes, intentions, and behaviors on campaign message exposure. Results confirm that level of sensation seeking was positively associated with greater message exposure. In addition, viewers reporting greater exposure were younger adolescents who indicated that they had poor family relations, promarijuana attitudes, and friends and family who used marijuana. Implications for designing future antimarijuana messages based on these findings are discussed. PMID- 11853209 TI - Investigating the physician-patient relationship: examining emerging themes. AB - By extending Millar and Roger's (1976) relational theory, this study examined how the physician-patient relationship was negotiated in the context of a clinical visit. Analysis of observational data obtained during the clinical visits of 1 female family practice physician revealed 5 themes central to the construction of her relationships with patients. The 5 themes included control, role negotiation, trust, health care commitment, and interrelated issues of time and money. PMID- 11853210 TI - Patients as partners, patients as problem-solvers. AB - This article reports our ongoing work in developing a model of health care communication called collaborative interpretation, which we define as a rhetorical practice that generates building blocks for a more complete and coherent diagnostic story and for a collaborative treatment plan. It does this by situating patients as problem-solvers. Our study begins with an analysis of provider-patient interactions in a specific setting-the emergency department (ED) of an urban trauma-level hospital- where we observed patients and providers miscommunicating in at least 3 distinct areas: over the meaning of key terms, in the framing of the immediate problem, and over the perceived role of the ED in serving the individual and the community. From our observations, we argue that all of these miscommunications and missed opportunities are rooted in mismatched expectations on the part of both provider and patient and the lack of explicit comparison and negotiation of expectations-in other words, a failure to see the patient-provider interaction as a rhetorical, knowledge-building event. In the process of observing interactions, conversing with patients and providers, and working with a team of providers and patients, we have developed an operational model of communication that could narrow the gap between the lay public and the medical profession-a gap that is especially critical in intercultural settings like the one we have studied. This model of collaborative interpretation (CI) provides strategies to help patients to represent their medical problems in the context of their life experiences and to share the logic behind their health care decisions. In addition, CI helps both patient and provider identify their goals and expectations in treatment, the obstacles that each party perceives, and the available options. It is adaptableto various settings, including short, structured conversations in the emergency room, extended dialogue between a health educator and a patient in a clinical setting, and group discussions in support groups, community groups, or health education classrooms. PMID- 11853211 TI - Women, menopause, and (Ms.)information: communication about the climacteric. AB - This research utilizes a communication perspective to examine the dissemination of information about menopause in terms of women's attitudes, beliefs, and knowledge. Specifically, this study uses a grounded theory approach (Glaser & Strauss, 1967) to explore the communicative processes of misinformation concerning women's lived experiences in relation to the climacteric. Five emergent themes extracted from premenopausal, perimenopausal, and postmenopausal women's discourse are identified and described through qualitative data analysis. Findings suggest that due to a lack of consistent communication, women are generally either unknowledgeable or misinformed about menopause and its related issues. Inaccurate information concerning a health-related experience that all women undergo has negative implications for women, their practitioners, and society. Moreover, a clearer understanding of women's experiences concerning menopause may enhance communication in physician-patient interactions (PPIs). PMID- 11853212 TI - Health literacy: an important issue for communicating health information to patients. AB - "Health literacy" refers to an individual's ability to read, understand, and use the information necessary to obtain adequate health care. Because so much health related information is provided to patients in written form, some degree of health literacy is essential. A review of the international medical and education literature was conducted to identify research on health literacy. Limited health literacy is a widespread problem in both non-industrialized and industrialized nations, and it is mostly related to deficient reading skills. Deficient reading skills are most prevalent among those with a limited educational background, but considerable research shows that limited reading skills exist even among those who have completed a formal education. Risk factors for limited literacy include poverty, membership in an ethnic minority group, advanced age, and other sociodemographic characteristics. Limited health literacy highly associated with poor health status, and literacy skills more accurately predict health status than education level, income, ethnic background, or any other sociodemographic variable. Individuals with limited literacy have higher rates of illness and more hospitalizations than individuals with more well-developed reading skills. Considerable effort has been directed at creating special health education materials for communicating with patients who have limited reading skills, but there is minimal evidence that these interventions have any effect on the health status of these individuals. In conclusion, poor health literacy is a common problem with important implications for health status. Limited data are available about how to best address the needs of patients with limited literacy skills. PMID- 11853214 TI - Endoscopic ligation for removal of stomach hyperptastic polyp: less risk or saving money? PMID- 11853213 TI - Sequential changes of gastric hyperplastic polyps following endoscopic ligation. AB - BACKGROUND: Endoscopic ligation has been extensively applied in the management of esophageal and gastric varices with or without bleeding. The varices are automatically eradicated through the use of ligation. However, whether avascular necrosis will occur in a gastrointestinal polyp when the base is ligated remains unclear. The aims of this pilot study were to investigate the sequential changes of gastric hyperplastic polyps following endoscopic detachable snare ligation and to determine the possibility of induction of avascular necrosis in these lesions following ligation. METHODS: Eleven patients with eighteen gastric hyperplastic polyps were treated with endoscopic detachable-snare ligation. The polyps were observed for 5 minutes and biopsies were then conducted. At 14 days after endoscopic ligation, follow-up endoscopies were performed to assess the outcome of the strangulated polyps. RESULTS: After being strangulated by the detachable snares, a majority of the polyps immediately congested (94%), and then developed cyanotic change (89%) approximately 4 minutes later. Pathological examination revealed severe venous congestion in the lamina propria of the strangulated polyps. On follow-up endoscopy 2 weeks later, all the snares had dropped off, and avascular necrosis occurred in sixteen polyps (89%). All of the polyps with avascular necrosis were detected to have developed cyanotic changes in initial endoscopy. No complications occurred during or following the ligation procedure. CONCLUSIONS: Most gastric hyperplastic polyps develop avascular necrosis following ligation by detachable snare. Cyanotic change is an important predictor of the outcomes of the lesions following endoscopic ligation. The application of this ligation technique in treatment of bleeding or non-bleeding gastrointestinal polyps deserves further investigation. PMID- 11853216 TI - Tele-emergency medicine: the evaluation of Taipei Veterans General Hospital and Kinmen-Granite Hospital in Taiwan. AB - BACKGROUND: The purpose of the project was to establish computer networks between selected hospitals through high-speed communication and high power computer processing to electronically exchange medical information and to conduct clinical examination and consultation. Quality medical services can thus be provided to patients in the remote rural areas such as villages and small towns in the mountains, on the coasts and islets away from Taiwan. It also intended to facilitate continuing education for doctors in those areas. This study evaluates telemedicine between Taipei-Veterans General Hospital and Kinmen-Granite Hospital. METHODS: Patients were chosen from 1996-7 to 1997-6. The evaluation criteria included consulting quality, satisfaction of the doctors, benefits for the patients, and the charge being rendered. RESULTS: The results of evaluation for telemedicine between Taipei-Veterans General Hospital (VGHTPE) and Kinmen Granite hospital (GH) are as follows: 93.0% doctors used telemedicine to seek a second opinion. After teleconsultation, the ratio of the patients showing cooperation was over 80%. Over 98% doctors thought telemedicine system helpful. The doctors in VGHTPE are more satisfied with the facility than local doctors in Kinmen. CONCLUSIONS: The clinical evaluation of the telemedicine showed positive results. It can be a useful tool to facilitate on-job training and education Tele emergency medicine. PMID- 11853215 TI - Modulation of Na, K-ATPase activity by immunoglobulins--III. Influence of an ATPase activity regulating agent "Marina" on Na, K-ATPase activity in patients with various chronic disturbances. AB - BACKGROUND: In some earlier work we have demonstrated that hypotonic mineral water can change the functional activity of Na, K-ATPase both in vitro and in vivo. The current investigation was aimed at measuring of the functional activity of ATPase in erythrocytes from blood samples of patients before and after treatment with the artificially mineralised water "Marina". METHODS: "Marina"--an artificial mineral water containing K+ and Mg2+ ions, was investigated by double blind method. Treatment lasted 7 days, daily intake of "Marina" was 4 x 200 ml before meals. Na, K-ATPase activity in erythrocytes and a number of clinical indices was determined before and after the treatment. RESULTS: After the intake of the preparation "Marina" the normal activity pattern of the Na, K-ATPase in chronic active hepatitis or atherosclerosis patients was restored. The restoration of the Na, K-ATPase activity brought about the normalization of vital clinical indices. CONCLUSIONS: The summarized data from the in vitro and in vivo experiments point to formation of Na, K-ATPase and immunoglobulin complexes as possible cause of cell disfunction and source of organic disturbances. PMID- 11853217 TI - Use factor analysis to identify difficulties in implementing a case payment reimbursement system in Taiwan. AB - BACKGROUND: Taiwan's National Health Insurance Program (NHIP) began on March 1, 1995. The major purpose of National Health Insurance is to remove financial barriers to care and enhance accessibility to comprehensive health care for all citizens in Taiwan. However, due to the rapid rise in medical expenses and unanticipated accumulation of overdue premiums, the program went into debt in 1999. In response to this financial crisis, the Bureau of National Health Insurance proposed to change hospital financing from a cost-based to a case-based reimbursement system. The major purpose of this study is to identify the difficulties in implementing a case payment reimbursement system in Taiwan. METHODS: This study was conducted in four stages: structured interviewing, questionnaire development and testing, surveying, and data analysis. In this study, the sampling surgeons were selected based on availability from 7 teaching hospitals and 10 regional hospitals. RESULTS: The results of factor analysis indicated that a five-factor structure has emerged in the use of the newly developed survey administered to 372 surgeons in Taiwan. These five factors accounted for approximately 97% of the overall variance. The alpha coefficient for this 22-item scale was 0.914. The researchers concluded that these five factors are considered evidence of adequate internal consistency for use in measuring difficulty in the implementation of case payment. CONCLUSIONS: These five factors were named as follows: perceived barriers to compliance, perceived barriers to quality assurance, perceived financial pressure, perceived threats to physicians' autonomy, and inadequate allowance for patient severity. Policy recommendations including annual revision of coding system, physician education, and hospital specialization were also made to address the perceived difficulties identified in this study. PMID- 11853218 TI - Combined innominate artery reconstruction and coronary artery bypass grafting. AB - Two patients having significant coronary artery disease with innominate artery near-total occlusion presented neurological deficit of syncope events or cerebellar and brain stem infarct. Both of them were successfully treated with one-stage reconstruction combined with aorto-carotid-subclavian bypass and coronary artery bypass grafting (CABG). While it could not be over-emphasized how to protect both myocardium and cerebrum during CABG, cerebral perfusion through the reconstructed carotid bypass graft is the key maneuver during cardiac arrest and moderate hypothermia. Hypoperfused cerebral hemispheres were both improved extensively in the follow-up angiography. The absence of cerebral deficit and the free from coronary angina suggested that surgical technique to combine innominate with coronary artery surgery is feasible with acceptable mortality and morbidity rate. PMID- 11853219 TI - Should simultaneous revascularization be recommended for patients with concomitant innominate artery disease and coronary artery disease? PMID- 11853220 TI - Abdominal vascular injury during lumbar disc surgery. AB - Major vascular injury is an unusual but well-recognized complication of vertebral disc surgery. Isolated arterial laceration is the most common type of this vessel injury in lumbar spine surgery, with early manifestation due to retroperitoneal hemorrhage. Two cases are described that illustrate the full spectrum of acute manifestation of such injuries. Two cases of acute hemorrhage due to arterial trauma were seen; one mortality case was recognized during the operation and one salvaged in the recovery room. In both cases unstable perioperative hemodynamics and postoperative distended abdomen were observed. It is the purpose of this paper to report two cases and to discuss the morbid anatomy, diagnosis of such vascular injuries and anesthetic handling of retroperitoneal hemorrhage. For anesthesiologists who are also drill workers while doing lumbar spinal or epidural anesthesia, these rare catastrophes remind us to pay special attention to the vertebral vascular (not only skeletal) anatomy. PMID- 11853221 TI - Resection of triple synchronous tumors--gastric adenocarcinoma, gallbladder adenocarcinoma and stromal tumor of the stomach. AB - We herein report a rare case of triple synchronous tumors: gastric adenocarcinoma, gallbladder adenocarcinoma and stromal cell tumor of stomach, which were resected in the same operation. A 72-year-old male patient suffered from poor appetite and epigastric pain that radiated to his back for one month. Upper gastrointestinal endoscopy showed a gastric ulcer measuring 1.5 cm at the prepyloric area, which was proven to be adenocarcinoma by biopsy. A second tumor was found in the gallbladder, measuring 3 cm, by ultrasonography. On computed tomography, a third tumor about 2.5 cm in size was recognized posterior to the gastric high body and anterior to the pancreas. Given the presence of three tumors, he underwent a radical subtotal gastrectomy with Billroth-II gastrojejunostomy and simultaneous cholecystectomy. Pathological findings revealed gastric adenocarcinoma, gallbladder adenocarcinoma and stromal cell tumor of the stomach. The postoperative course was smooth. PMID- 11853222 TI - Endovascular aortic graft exclusion of abdominal aortic aneurysm. AB - Endovascular repair of abdominal aortic aneurysms has evolved dramatically within the past few years. This study reports the first successful case in Taiwan area of infrarenal abdominal aortic aneurysm treated with endovascular aortic bifurcational stent-graft. Minimally invasive surgery leading to early recovery, mobilization and shortened hospital day was promising. In addition, there were no local, remote or systemic complications during a 18-month follow-up. Thus, the procedure is an attractive alternative in its potential to reduce morbidity and mortality associated with open surgical repair, especially for patients who are not surgical candidates because of comorbidities. Careful patient selection and more experiences with refined endograft models will elucidate the feasible alternative to conventional surgical repair. PMID- 11853223 TI - Experimental atherosclerosis: a historical overview. AB - Almost one-hundred years ago the first evidence of experimental atherosclerosis was reported. Over the past century, significant advances have been made in the development of animal models of human coronary artery disease. In this minireview, induction of atherosclerotic lesions in several animal models including rodents (mice, rabbits, rats, hamsters, guinea pigs), avian (pigeons, chickens, quail), swine, carnivora (dogs, cats), and non-human primates is discussed. The limitations and advantages of the animal models of atherosclerosis have been summarized. The transgenic/knockout animal models have greatly enhanced our understanding of atherosclerosis. Compared to wild-type counterparts, the knockout/transgenic animals develop atherogenesis faster without a need for a highly atherogenic diet. Although almost all investigations support a causal role for increased plasma cholesterol levels in the development of atherosclerotic vascular disease, an increasing body of evidence indicates serious invqlvement of other factors including oxidative stress, inflammation, infection and other emerging risk factors. PMID- 11853224 TI - Differential expression of decay-accelerating factor isoforms in the digestive tract of guinea pig. AB - Decay-accelerating factor (DAF, CD55), one of the membrane-binding regulators of complement activation, prevents host cells from the autologous complement mediated attack. However, specific localization of DAF in the different layers of the digestive tract has not been defined. Using a crypt isolation procedure, we separated epithelium from the remnant stromal tissue and assessed the expression of DAF isoforms in the two layers in the guinea pig digestive tract. Both Northern hybridization and immunohistochemistry revealed DAF was preferentially expressed in the small intestinal epithelium compared to the stomach and colon. Reverse-transcriptase PCR demonstrated that the glycosyl-phosphatidylinositol (GPI)-anchored isoforms are predominant over the transmembrane (TM) isoforms in all digestive epithelium. However, there was no difference between GPI-anchored type and TM type in the stromal tissue. In conclusion, DAF was found to be primarily expressed in the small intestinal epithelium with isoforms differing between the epithelium and stromal tissue, suggesting differential roles in the guinea pig digestive tract. PMID- 11853225 TI - Isolation of a new heterodimeric lectin with mitogenic activity from fruiting bodies of the mushroom Agrocybe cylindracea. AB - From the dried fruiting bodies of the mushroom Agrocybe cylindracea a heterodimeric lectin with a molecular weight of 31.5 kDa and displaying high hemagglutinating activity was isolated. The molecular weights of its subunits were 16.1 kDa and 15.3 kDa respectively. The larger and the smaller subunits resembled Agaricus bisporus lectin and fungal immunomodulatory protein from Volvariella volvacea respectively in N-terminal sequence. The lectin was adsorbed on DEAE-cellulose in 10 mM Tris-HCl buffer (pH 7.4) and was eluted by the same buffer containing 150 mM NaCl. It was adsorbed on SP-Sepharose in 10 mM NH4OAc (pH 4.5) and eluted by approximately 0.19 M NaCl in the same buffer. The lectin was obtained in a purified form after the mushroom extract had been subjected to (NH4)2SO4 precipitation and the two aforementioned ion exchange chromatographic steps. The lectin exhibited potent mitogenic activity toward mouse splenocytes. The hemagglutinating activity of the lectin was inhibited by lactose, sialic acid and inulin. PMID- 11853226 TI - Naturally occurring human autoantibodies recognize a fetal brain antigen identified as microtubulus associated protein 1B. AB - We have recently reported naturally occurring autoantibodies against a large fetal brain antigen (FBA). Now we describe the process of purification and identification of this particular FBA. The brains of newborn rabbits were solubilized and purified with preparative gel electrophoresis. The protein fractions were concentrated and desalted and the fractions were tested by a known positive serum. On membrane digestion of the FBA-band gave a twelve amino acid sequence that resulted in best identity score for mouse, rat and human microtubule-associated protein (MAP) 1B: a member of the microtubule-associated protein family. Monoclonal anti-MAP1B recognized a band in immunoblots of the brain homogenate and of the partially purified fractions with the same electrophoretic mobility as that recognized by a known anti-FBA positive serum. When adult rabbit brain was used as an antigen, the anti-MAP1B failed to recognize any bands on immunoblots. MAP lB has not been previously known as an autoantigen, even though many structural proteins of the neuronal cytoskeleton are known to be targets of naturally occurring autoantibodies. MAP 1B is a functionally important regulatory protein in the developing brain; thus autoantibodies against MAP1B may affect the normal development. PMID- 11853227 TI - Biological properties of a new fluorescent biphalin fragment analogue. AB - Previous studies of structure-activity of biphalin defined fragments which expressed the full biological potency of the parent compound. The most simple fragment was Tyr-D-Ala-Gly-Phe-NH-NH<--X, where X=Phe, but it also could be other hydrophobic amino acids. This paper presents data that replacement of the phenylalanine with a dansyl (X=DNS) groups gives an analogue (AA2016) that fully preserves the high affinity of the initial analogue for both mu and delta opioid receptors. In the tail flick test in rats, intrathecal injection of the compound produces strong antinociception, comparable to the parent biphalin. Because AA2016 contains a strong fluorescent group, it can be a very useful tool for prospective studies in vivo, including biological barrier permeability, tissue distribution, metabolism and receptor-ligand complex formation. PMID- 11853229 TI - Methemoglobin contributes to the growth of human tumor cells. AB - Methemoglobin (metHb) has been reported to be present in areas surrounding solid tumors. The effects of human metHb on the growth of one human hepatocellular carcinoma cell line and one human glioma cell line that simply replicate in Ham's nutrient mixture F12 (F12) were investigated. MetHb, depending on its concentration, stimulated or inhibited the in vitro growth of both cancer cell lines. The stimulatory or inhibitory effect was due to the release of hemin from metHb, which was recognized by its characteristic light absorption spectrum. The possibility of metHb or hemin acting initially through a 3', 5'-cyclic guanosine monophosphate- (cGMP-) or prostaglandin E2- (PGE2-) mediated pathway to enhance cell growth was excluded. Ferric iron derived from the catabolic degradation of hemin increased cell growth, whereas biliverdin (Bv) and its reduction product, bilirubin (Br), decreased cell growth. Hemoglobin oxidized to metHb in conditions found in tumors showing neovascularization and hemorrhage may contribute significantly to increased proliferation of cancerous cells. PMID- 11853228 TI - Luffangulin, a novel ribosome inactivating peptide from ridge gourd (Luffa acutangula) seeds. AB - A ribosome inactivating peptide, with an N-terminal sequence exhibiting pronounced similarity to that of the 6.5 kDa-arginine/glutamate-rich polypeptide from Luffa cylindrica seeds, was isolated from seeds of a closely related species, the ridge gourd Luffa acutangula. The 5.6 kDa-peptide designated luffangulin inhibited cell-free translation with an IC50 of 3.5 nM but lacked inhibitory activity toward HIV-1 reverse transcriptase. It was similar to luffaculin, the 28 kDa ribosome inactivating protein in being unadsorbed on DEAE cellulose and adsorbed on Affi-gel blue gel. On CM-cellulose luffangulin and luffaculin appeared as two adjacent peaks. PMID- 11853230 TI - Reduced expressions of inducible nitric oxide synthase and cyclooxygenase-2 in vascular smooth muscle cells of stroke-prone spontaneously hypertensive rats. AB - Inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2 are expressed in vascular smooth muscle cells stimulated with interleukin-1beta (IL-1beta), resulting in the production of nitric oxide (NO) and prostaglandins (PGs) such as PGI2. The iNOS and COX-2 proteins and their mRNA expressions in cultured vascular smooth muscle cells isolated from 6-7 week-old stroke-prone spontaneously hypertensive rats (SHRSP) were compared with those in the cells isolated from age matched normotensive Wistar Kyoto rats (WKY). The IL-1beta-induced NO production and iNOS expression in vascular smooth muscle cells of SHRSP were significantly lower than those in cells of WKY. Similarly, PGI2 production and COX-2 expression were significantly lower in vascular smooth muscle cells of SHRSP than WKY, whereas there was no difference in the COX-1 expression. There were no significant differences in iNOS and COX-2 mRNA expressions between the two strains, suggesting that these protein expression may be reduced at the post transcriptional level in cells of SHRSP. These results further suggest that the reduction of iNOS and COX-2 expressions in vascular smooth muscle cells may have relevance to the pathophysiology in SHRSP. PMID- 11853231 TI - The role of kinin B1 in the plasma extravasation of carrageenin-induced pleurisy. AB - The role of des-Arg9-bradykinin (des-Arg9-BK) and kinin B1 receptor in the plasma extravasation of rat carrageenin-induced pleurisy was investigated employing B1 receptor agonist and antagonists and kininogen-deficient rats. Expression of the B1 receptor mRNA in pleura was induced from 3 to 5 h after the injection of carrageenin into the pleural cavity of Sprague-Dawley rats. Exogenous injection of des-Arg9-BK into the pleural cavity provoked a significant increase in plasma extravasation in 5 h carrageenin-induced pleurisy, but not in 20 min kaolin induced pleurisy. The level of immunoreactive des-Arg9-BK in the exudate of 5 h carrageenin-induced pleurisy was higher than that of bradykinin (BK). Administration of the B1 receptor antagonists, des-Arg9-[Leu8]-BK or des-Arg9-D Arg-[Hyp3, Thi5, D-Tic7,Oic8]-BK significantly reduced the exudation rate. However, intrapleural administration of des-Arg9-BK to plasma kininogen deficient. Brown Norway-Katholiek rats did not result in a further increase in the plasma extravasation. In conclusion, endogenously generated des-Arg9-BK could contribute to the plasma extravasation in carrageenin-induced pleurisy via mediation of the inducible B1 receptor. PMID- 11853232 TI - Mast cell dynamics and involvement in the development of peritoneal adhesions in the rat. AB - Mast cells have been implicated in the ethiopathology of post-operative peritoneal adhesions. However an evaluation of their role in this condition is missing. Adhesions were induced in rats using small intestinal scraping. These rats or rats injected ip with either Stem Cell Factor (SCF) or nedocromil sodium or compound 48/80 (day 0-20) were sacrificed for grading of peritoneal adhesions, for evaluating mast cells and inflammatory cells in adhesions and peritoneal lavage (histochemical staining) and for histamine content (peritoneal lavage, radioenzymatic assay) on days 1-21. Mast cell sonicate was added to intestinal fibroblast and their proliferation was assessed (cell counting). All the rats developed adhesions (day 1) and after 3 days the adhesion score remained constant. Early adhesions were avascular and made of fibrinous exudate containing many mast cells. Thereafter adhesions became denser, and the number of stainable mast cells decreased and then stabilized. On the first few days, inflammatory cells in the peritoneal lavage increased while mast cells and histamine content were significantly reduced indicating their activation. Injection of SCF for 1 week slightly increased peritoneal adhesion formation while nedocromil sodium reduced their development. Compound 48/80 had no significant influence. Addition of mast cell sonicate to normal intestine or to peritoneal adhesion fibroblasts resulted in a significant increase of fibroblast proliferation. In conclusion, mast cell presence correlated with the establishment of peritoneal adhesions, and their pharmacological modulation influenced adhesion formation. In vitro mast cell induced fibroplasia. Therefore, mast cells have a profibrogenic role in this model of peritoneal adhesions. PMID- 11853233 TI - Histamine pharmacokinetics in tumor and host tissues after bolus-dose administration in the rat. AB - In this study, we estimated interstitial histamine concentrations in normal and malignant tissues after a single intravenous (i.v.) injection of 0.5 mg/kg histamine dihydrochloride in the rat. The microdialysis technique was used to collect interstitial fluid from subcutis, liver and a NGW adenocarcinoma. Histamine was absorbed with equal efficiency to all tissues (t 1/2 AB 3.9-7.7 minutes) but maximum concentration (Cmax; nmol/l) of histamine was higher in liver (2,388 +/- 357) than in subcutis (951 +/- 125) (p < 0.01) and subcutaneous tumor (523 +/- 140) (p = 0.01) and, moreover, Cmax in liver tumor (1,752 +/- 326) was higher than in subcutaneous tumor (p = 0.01). The tl/2 elimination was significantly longer in subcutis and subcutaneous tumor than in liver and liver tumor. Area under the curve (AUC; mmol-min/l) for histamine was significantly lower in subcutaneous tumor (9.8 +/- 2.3) than in liver (17.6 +/- 1.9) (p = 0.03) and liver tumor (15.8 +/- 1.8) (p = 0.03). Local tissue blood flow as assessed by the 14C-ethanol method was not significantly altered by the histamine administration. In conclusion, after an i.v. injection of histamine dihydrochloride a higher maximum concentration and AUC of histamine was reached in liver and liver tumor than in subcutaneous tissues. PMID- 11853234 TI - A protocol for polymerase chain reaction detection of Enterococcus faecalis and Enterococcus faecium from the root canal. AB - AIM: The present study was set up to develop a protocol for detection of Enterococcus faecalis and Enterococcus faecium from the root canal. METHODOLOGY: A collection of type strains and clinical isolates ol E. faecalis and faecium was used. Specific polymerase chain reaction (PCR) primers targeted against the 16S/23S rDNA intergenic region were used and PCR reactions were set up. PCR products were run on TBE-agarose gel and analysed. The sensitivity of the PCR systems was studied using serial dilutions of (i) bacterial DNA and (ii) bacterial cells from E. faecalis. The specificity of the identification was tested against closely related species. RESULTS: All strains of E. faecalis and E. faecium produced identical amplicon profiles composed of two major bands corresponding to sizes of 320 and 420 bp. When amplifying DNA of higher purity, a third band of 600 bp became evident as well. Closely related species demonstrated single bands of various sizes and were easily distinguished from enterococci. The detection level of DNA from serial dilutions of DNA was 10(-13) g. The DNA extraction protocol from bacterial cell suspensions resulted in a detection level of 10 bacterial cells per sample. CONCLUSIONS The present study demonstrated a good potential for using PCR technology in the detection of F. faecalis and E. faecium from root canal samples. With a high specificity the methodology was able to detect 10 cells of E. faecalis. PMID- 11853235 TI - Occlusal loading of EBA and MTA root-end fillings in a computer-controlled masticator: a scanning electron microscopic study. AB - AIM: The aim of this investigation was to assess the marginal adaptation of Super EBA (EBA) and Pro Root MTA (MTA) root-end fillings and the occurrence of microcracks in resected root-ends of extracted teeth before and after occlusal loading for a five-year equivalent period in a computer-controlled masticator. METHODOLOGY: Twenty-four molar teeth were root-filled using lateral condensation and their root-ends resected. Root-end cavities were prepared and either EBA or MTA root-end fillings were placed. All surgical procedures were completed in vitro under an operating microscope. Replicas taken of the root-end fillings and resected root-ends were examined using an SEM before and after they were subjected to in vitro chewing cycles in a computer-controlled chewing simulator for the equivalent of 5 years. Marginal adaptation and integrity of the two root end filling materials were evaluated and the presence of microcracks recorded. RESULTS: Before loading, a continuous margin was observed with 99.4 +/- 2.4% of EBA and 99.2 +/- 1.6% of MTA root-end fillings. After loading, the percentage of continuous margin decreased to 93.1 +/- 6.6% and 98.9 +/- 3.2% for EBA and MTA, respectively. After occlusal loading, 39.5% of the EBA root-end fillings were overfilled, whilst 52.6% of the MTA root-end fillings were underfilled. In total, 12.5% of the resected root-end surfaces showed microcracking before loading and 25% after occlusal loading. CONCLUSIONS: Both EBA and MTA displayed excellent marginal adaptation before masticatory loading. After loading, the amount of continuous margin for both root-end filling materials decreased slightly but was still high. PMID- 11853237 TI - A comparative study of root canal preparation using Profile .04 and Lightspeed rotary Ni-Ti instruments. AB - AIM: The purpose of this Study was to compare several parameters of root canal preparation using two different rotary nickel-titanium instruments: ProFile .04 (Dentsply/ Maillefer, Ballaigues, Switzerland) and Lightspeed (Lightspeed Technology Inc., San Antonio, TX, USA). METHODOLOGY: Fifty extracted mandibular molars with root canal curvatures between 20 degrees and 40 degrees were divided into two similar groups having equal mean curvatures. The teeth were then embedded into a muffle system as described by Bramante et al. (1987) and modified by Hulsmann et al. (1999b). All root canals were prepared using ProFile .04 or Lightspeed Ni-Ti instruments to size 45 following the manufacturers' instructions. The Lightspeed system was used in a step-back technique: ProFile .04 instruments were used in a crown-down technique. The following parameters were evaluated: straightening of curved root canals (superimposition of pre- and postoperative radiographs), postoperative root canal diameter (superimposition of pre- and postoperative photographs of root canal cross-sections), safety issues (file fractures. perforations, apical blockages, loss of working length) (protocol), cleaning ability (SEM-evaluation of root canal walls using a five score system for debris and smear layer), and working time (protocol). Statistical analysis was performed using the Wilcoxon test (P < 0.05) for straightening, and Fisher's exact-test (P < 0.05) for comparison of cross sections, for comparison of contact between pre- and postoperative diameter, root canal cleanliness and working time. RESULTS: Both Ni-Ti systems maintained curvature well; the mean degree of straightening was less than 1 degrees for both ProFile .04 and for Lightspeed with no statistical significance between the groups. Most procedural incidents occurred with Profile .04 instruments (three fractures), Lightspeed preparation was completed without instrument fractures. Loss of working length, perforations or apical blockage did not occur with either instrument. Following preparation with Profile.04. 64.0% of the root canals had a round, 30.7% an oval, and 5.3% an irregular cross-section, Lightspeed preparation resulted in a round cross-section in 41.3% of cases, an oval shape in 45.3% of cases: 13.3% of cases had an irregular cross-section. No significant differences were found between the two systems. Lightspeed instruments enlarged the root canal more uniformly with no specimen showing 50% or more contact between pre- and postoperative diameter. The difference was statistically significant only for the coronal third of the root canals (P = 0.032). Mean working time was significantly shorter for Profile .04 (105 s) than for Lightspeed (140 s) (P = 0.02). For debris removal Lightspeed achieved the best results (68% scores 1 and 2), followed by Profile .04 (48.4%) with no significant differences between the systems. The results for remaining smear layer were similar: the lowest amount of smear layer on the root canal walls was found after preparation with Lightspeed (30.7% scores 1 and 2). followed by Profile.04 (23.1%). In the coronal third of the root canals Lightspeed performed significantly better than Profile .04 (P = 0.029): in the middle and apical third the differences were not significant. CONCLUSIONS: Both systems under investigation respected original root canal curvature and were safe to use. Both systems can be recommended for clinical use. PMID- 11853236 TI - Quantitative analysis of substance P, neurokinin A and calcitonin gene-related peptide in pulp tissue from painful and healthy human teeth. AB - AIM: The purpose of this study was to investigate the levels of substance P (SP), neurokinin A (NKA) and calcitonin gene-related peptide (CGRP) in painful and healthy human dental pulps. METHODOLOGY: Forty-six samples of pulp tissue were collected from extracted or endodontically treated painful teeth and 20 from clinically healthy teeth extracted for orthodontic reasons. All pulp samples were boiled in 0.5 m acetic acid for 10 min, centrifuged and the supernatant collected. SP, NKA and CGRP levels were measured using radioimmunoassay. RESULTS: Substance P and CGRP were present in all samples and NKA was detected in 96% of the pulps. CGRP was present in much higher concentrations than SP and NKA in both painful and non-painful teeth. The painful teeth had significantly higher concentrations of SP (P = 0.02), NKA (P < 0.001) and CGRP (P = 0.03) than non painful teeth. The concentration of CGRP was significantly higher in the pulps of smokers compared with non-smokers (P = 0.02). CONCLUSIONS: Elevated levels of these neuropeptides in pulps from painful teeth indicate that they may play an important role in the process of pulpal inflammation and pain. Further investigation of the association between these neuropeptides and pulpal status may help to improve our understanding of pulpal inflammation and dental pain. PMID- 11853238 TI - In vitro cytotoxicity of a composite resin and compomer. AB - AIM: This work was designed to investigate the potential cytotoxicity of two of the newer dental restorative materials. Spectrum composite resin and Dyract AP compomer. METHODOLOGY: Cultured human endothelial cells (ECV-304) were exposed to each of the restorative materials through a 70-microm dentine barrier to simulate the in vivo clinical situation. Cell viability was measured by the MTT (3-[4,5 dimethylthiazol-2-yl]-2,5 diphenyltetrazolium bromide) assay and lactate dehydrogenase release assay. The effects of different extents of light-curing were also examined by microscopic examination of stained human promyelocytic leukemia cells (HL-60). Caspase-3 activation was determined as a measure of apoptotic cell death. RESULTS: Assessment of cellular viability indicated that both materials cause cell death, with Spectrum being the more toxic. The cytotoxicity was considerably increased in the absence of the dentine barrier. Direct exposure to Spectrum for 12 h resulted in the death of 69% of the cells after full light-curing (78% of total death was by apoptosis) and 96% after partial light-curing (73% of total death was by necrosis). Assessment of caspase activation, in the absence of the dentine barrier, showed that longer curing times resulted in an increase in the proportion of the cells dying through apoptosis, rather than necrosis, for both materials tested. CONCLUSIONS: These results indicate the restorative materials to be potentially toxic, particularly if the degree of light-cure is inadequate. PMID- 11853239 TI - Root and canal morphology of Thai mandibular molars. AB - AIM: To investigate the root and canal morphology of 351 mandibular permanent molars collected from an indigenous Thai population. METHODOLOGY: The cleaned teeth were accessed, the pulp dissolved by sodium hypochlorite under ultrasonication and the pulp system injected with Indian ink. The teeth were rendered clear by demineralization and immersion in methyl salicylate. The following observations were made: (i) number of roots and their morphology; (ii) number of root canals per tooth; (iii) number of root canals per root; (iv) root canal configuration in each root using Vertucci's classification, with additional modifications; and (v) presence and location of lateral canals and intercanal communications. RESULTS: Of 118 mandibular first molars, 13% had a third disto lingual root. In three-rooted teeth, 80% of the main distal roots and 100% of the disto-lingual roots had type 1 canal systems. Of 60 mandibular second molars, 10% had C-shaped roots, the majority of which had type 1 (33%) or type IV (33%) canal systems. Of 173 third molars, 68% had two separate roots, 20% had fused roots and 11% had a single C-shaped root; the majority had two canals (61%). CONCLUSIONS: This study suggests that Thai molars exhibit features of both Caucasian and Chinese teeth. PMID- 11853240 TI - Incidence of root fractures and methods used for post removal. AB - AIM: This study was undertaken to determine the incidence of root fractures during post removal, as well as the methods used and the success rates achieved with various post removal devices and techniques. METHODOLOGY: Patient records from a specialist endodontic practice were examined. A total of 1600 teeth from which posts were removed were identified. These teeth had no preoperative signs or symptoms to suggest root fractures. The incidence of root fractures was determined and a subset of 234 patient records were examined in detail to determine the methods used to remove different types of posts. RESULTS: Only one tooth fractured during post removal (0.06%) and all posts were successfully removed, typically in about 3 min. Most cast posts were removed with an Eggler post remover but some dislodged when the crown was removed; ultrasonics was used occasionally. Most parallel-sided posts were removed with ultrasonics and threaded posts were unscrewed. The Masserann kit and ultrasonics were combined to remove fractured cast posts and some parallel-sided posts. CONCLUSIONS: This study indicates that, with good case selection, post removal is a predictable procedure. If appropriate techniques and devices are used for the particular type of post being removed, then root fracture is a rare occurrence. PMID- 11853241 TI - Non-surgical root canal treatment of dens invaginatus type 2 in a maxillary lateral incisor. AB - AIM: To describe the clinical management of an unusual dens invaginatus type 2. SUMMARY: A case of dens invaginatus in a maxillary lateral incisor with a periapical lesion is reported. The patient presented with pain and localized swelling. Despite the complex anatomy and diagnosis of dens invaginatus, non surgical root canal treatment was performed successfully. Key learning points Dens invaginatus may be presented in many forms, and the aetiology of this phenomenon is not fully understood. Due to abnormal anatomical configuration, dens invaginatus presents technical difficulties in its clinical management. Non surgical root canal treatment can be performed successfully. PMID- 11853242 TI - Endodontic treatment of teeth associated with a large periapical lesion. AB - AIM: To report the healing of a large periapical lesion following non-surgical root canal treatment. SUMMARY: A 12-year-old male with a large periapical lesion around the apices of his mandibular incisors is described. The lesion formed after trauma to the mandibular anterior teeth 5 years previously. During root canal treatment, chlorhexidine gluconate was used for irrigation, and calcium hydroxide was used both for the intracanal dressing and as a base of the root canal sealer. Periapical healing was observed 3 months after obturation and continued at the 12-month review. Key learning points Root canal treatment, including the use of chlorhexidine gluconate and calcium hydroxide for infection control, led to substantial healing of a large periapical lesion. This report confirms that large periapical lesions can respond favourably to non-surgical treatment. PMID- 11853243 TI - Oral manifestations in 22q11 deletion syndrome. AB - OBJECTIVES: The aims of this study were to investigate and describe oral manifestations in 22q11 deletion syndrome, and to relate the findings to medical conditions. DESIGN: Cross-sectional. SAMPLE AND METHODS: Fifty-three consecutive patients from Sweden referred to the Sahlgrenska University Hospital during a 3 year period were included, median age 8 yrs (range 2-43; mean age 11.09). All but six patients were children 3-19-years-old. The patients were examined concerning oral mucosa, dental anomalies, dental caries, occlusal development, and eruption of the teeth. The clinical findings were compared to medical data and case history. RESULTS: Dental anomalies were registered in high numbers. Enamel hypoplasia was found in 16 patients. In 13 cases this was documented in primary teeth, of which 10 patients had symmetrical and chronological defects. Enamel hypomineralization was found in 23 patients and was equally common in both primary and permanent teeth. The use of computerized inductive analyses revealed that enamel hypoplasia was associated with medical conditions like preterm birth and congenital heart malformation while hypomineralization was associated with more diffuse conditions like frequent infections. Hypodontia was registered in seven patients, while eight had aberrant tooth shape, and nine patients presented delayed tooth eruption. The patients had an average of 4.6 carious or filled teeth and the oral health was assessed as impaired in 15 patients who had severe dental caries (5-18 carious teeth or multiple active incipient caries lesions). CONCLUSIONS: In 22q11 deletion syndrome the oral cavity is affected by anomalies in dental enamel, tooth shape, numbers of teeth, and eruption. Dental health problems due to caries are common. This is of special importance as patients with 22q11 deletion syndrome frequently present with congenital heart malformations and immunological problems. PMID- 11853244 TI - Prevalence of salivary Streptococcus mutans in mothers and in their preschool children. AB - OBJECTIVES: To establish the prevalence and possible relationship of oral Streptococcus mutans colonization in mother-child pairs. DESIGN AND SETTING: An analytical cross-sectional study was carried out at a well-baby evaluation clinic held at the Public Dental Clinic, Varberg, Sweden. SAMPLE AND METHODS: Two hundred preschool children, 100 of which were 18 months old and the remaining 100 were 3 years old, and their mothers attended the clinic. All mothers were interviewed and their children's medical history, oral hygiene routines and dietary habits established. All children were clinically examined. The presence and level of S. mutans was estimated in the mother-child pairs with the aid of the Strip mutans chair-side test. RESULTS: Nearly 50% of mothers exhibited high levels of salivary S. mutans, prevalence among the 18-month- and 3-year-olds was 30% and 42%, respectively. A statistically significant (P < 0.01) mother-child relationship was found; a greater presence in mothers led to a higher number of children found harbouring the bacteria. Logistic regression analysis found that high maternal S. mutans levels (P < 0.001), daily sweet intake (P < 0.01) and sugary drinks in feeding bottles (P < 0.05) were significant factors for S. mutans colonization in children. Absence of daily toothbrushing and use of feeding bottles at night failed to fit into the model. Caries prevalence (initial and manifest decayed surfaces) was significantly related to S. mutans colonization (P < 0.01). CONCLUSION: The results support the concept of vertical transmission (mother-child), emphasizing the importance of the dietary component, and justifying a primary preventive approach with targeted action directed at mothers with high levels of S. mutans colonization. PMID- 11853245 TI - Dental treatment, dental fear and behaviour management problems in children with severe enamel hypomineralization of their permanent first molars. AB - OBJECTIVES: To assess the demand for restorative treatment, dental anxiety and dental behaviour management problems among children with severe hypomineralization of their first molars (MIH). DESIGN: Case control study. SAMPLE AND METHODS: Data were compiled from the dental records of 32 9-year-old children with severe enamel hypomineralization of their first molars and from 41 controls of the same age group concerning dental health, a number of restorative treatments, use of local anaesthesia and clinical behaviour management problems (BMP). A questionnaire containing specific questions on children's experiences of dental care and the Children's Fear Survey Schedule-Dental Subscale (CFSS-DS), was answered by the parents. RESULTS: The children in the study group had undergone dental treatment of their first molars nearly 10 times as often as the children in the control group. Repeated treatments of these teeth at brief intervals were common. The dental treatment was often undertaken without the use of local anaesthesia, and BMP, and dental fear and anxiety (DFA) were more common than in the controls. CONCLUSIONS: Children with severe enamel hypomineralization of their first molars had had to undergo a considerable amount of dental treatment. It is reasonable to assume that experiences of pain and discomfort on repeated occasions were related to the occurrence of BMP in patients with MIH. An early treatment planning and prognostication based on increased knowledge of hypomineralized first molars is desirable. Local anaesthesia and other pain reducing techniques, e.g. sedation, should be used when treating these teeth. Extraction should be considered in cases of extensive disintegration of the crown, in cases of frequently repeated treatments or when pulpal symptoms are hard to cure. PMID- 11853246 TI - Temporo-mandibular joint kinetics and chewing cycles in children. A 3-year follow up. AB - OBJECTIVES: To report the temporo-mandibular joint (TMJ) kinetics and masticatory function in healthy children. DESIGN: Temporo-mandibular joint palpation and electrognathographic registrations of chewing cycles were repeated for 3 years in order to evaluate changes. SETTING: Healthy children without systemic pathologies, decayed cavities and previous dental treatment. SAMPLE AND METHODS: Electrognathographic (EGN) registration of masticatory cycles and TMJ palpation were carried out on 52 patients (mean age: 5 years 8 months, range: 5 years 1 month, 6 years 8 months), by two university researchers, once a year for 3 consecutive years. TMJ palpation, differentiated TMJ synchronism (simultaneous bilateral opening movement) and TMJ asynchronism (not simultaneous bilateral opening movement), TMJ subluxation and click were observed. Electrognathographic registrations differentiated normal and abnormal jaw chewing cycles, and narrow and large cycles. RESULTS: Temporo-mandibular joint asynchronism was evident in 34 of 52 patients in the primary dentition, in 42 of 52 patients after the eruption of the first permanent molar, and in 31 of 52 patients after the eruption of the permanent incisors. TMJ subluxation increased during the full period of observation. Three temporomandibular clicks appeared after the eruption of the permanent incisors. Altered mastication was not always associated with TMJ disorders. CONCLUSIONS: In children, normal chewing cycles can coexist with occlusal discrepancies, cranio-facial growth and TMJ alterations. PMID- 11853247 TI - Marginal seal of sealant and compomer materials with and without enameloplasty. AB - OBJECTIVES: This in vitro study evaluated the quality of marginal seal and fissure penetration of a resin-sealant (Concise) and a compomer restorative material (F2000) with and without enameloplasty. SAMPLE AND DESIGN: Thirty extracted sound human permanent third molar teeth were divided into six groups of five and used in this in vitro study. METHODS: Occlusal surfaces of the teeth in different groups were either left intact or prepared using a #582 S diamond bur. Teeth were then restored with Concise and F2000 restorative materials using combinations of different surface treatments. Teeth were prepared for microleakage, then immersed in 5% methylene blue for 4 h, and subsequently sectioned into four bucco-lingual sections. Sections (20 per group) were examined using a stereomicroscope. The following parameters were evaluated: marginal leakage, marginal gap, voids and filling defects, as well as enamel fissure penetration. RESULTS: Kruskal-Wallis one-way ANOVA showed statistically significant differences between the groups for marginal leakage (P < 0.0001), marginal gap (P < 0.001), and enamel fissure penetration (P < 0.001). No statistically significant differences between the groups for voids and filling defects were detected. Non-parametric Tukey's multiple range test showed that microleakage of sealant with acid etch and tooth preparation and compomer with acid etch, bonding agent and tooth preparation were statistically significantly less than that of compomer with bonding agent and no tooth preparation (P < 0.0001) and compomer with bonding agent and tooth preparation (P < 0.01). CONCLUSIONS: Microleakage is similar when sealant with acid etching, and compomer with acid etching and bonding agent, are used with and without enameloplasty. Marginal gap is more evident in the compomer group with bonding agent and without enameloplasty. Enamel fissure penetration is better when enameloplasty and acid etching are used. PMID- 11853248 TI - Validation of a Facial Image Scale to assess child dental anxiety. AB - OBJECTIVE: To examine the validity of a scale that uses faces as an indicator of children's dental anxiety. SETTING: Department of Child Dental Health waiting room, Newcastle Dental Hospital. SUBJECTS AND METHODS: 100 children (aged 3-18 years) completed the Facial Image Scale (FIS) and the Venham Picture Test (VPT) in the dental hospital waiting room. RESULTS: A strong correlation (0.7) was found between the two scales, indicating good validity for the FIS. Findings also showed that a small, but significant, number of children are anxious in the dental context. CONCLUSION: The findings suggest that the FIS is a valid means of assessing child dental anxiety status in a clinical context. PMID- 11853249 TI - Delayed eruption of premolars with periodontitis of primary predecessors and a cystic lesion: a case report. AB - Apical periodontitis after pulp therapy in a primary tooth can cause delayed eruption of the permanent successor. A case of bilateral delayed eruption of mandibular premolars is presented. The patient. a 13-year-old girl, was referred by her dentist. Oral findings showed that the right first and left second primary molars were retained. Other premolars had erupted. An orthopantomogram revealed apical periodontitis, affecting both retained primary molars. The right first mandibular premolar was impacted against the alveolar bone and root of the second premolar, and there was a large cystic lesion in close association with the left second mandibular premolar. Both primary molars were extracted, and the cystic lesion was treated by marsupialization. Fenestration and traction were performed on the right first premolar. Correct tooth alignment was achieved with orthodontic appliances. If the problem had been detected earlier, treatment of the premolars might have been easier. Clinical and radiological follow-up, therefore, of primary teeth that have undergone pulp therapy procedures should be performed until eruption of succedaneous teeth. PMID- 11853251 TI - Dental disease indices and caries-related microflora in children with glycogen storage disease. AB - OBJECTIVE: To establish the levels of dental caries, bacterial dental plaque, gingivitis and caries-related microflora in children with glycogen storage disease (GSD). Patients with GSD are treated with regular intakes of glucose polymer and uncooked cornstarch to prevent hypoglycaemia. Dental health data are scarce. STUDY GROUP: The study group comprised 21 children with GSD attending the Great Ormond Street Hospital for Children. OUTCOME MEASURES: These included the number of decayed, missing and filled teeth, and surfaces in both the primary and permanent dentitions, plaque and gingivitis scores. Both plaque and saliva were collected from each child and cultured for Mutans streptococci, Lactobacilli and Candida. RESULTS: The study group included 13 boys and eight girls, aged from 2.7 to 15.5 years. Four of the 21 children had some caries experience. The mean dmft was 0.5 and the mean DMFT, 0.06. Mean plaque and gingivitis scores were 4.8 and 5.9, respectively, for plaque and gingivitis adjacent to the primary teeth, and 11.6 and 12 for those related to permanent teeth. CONCLUSIONS: Only a small proportion of the children had caries experience but most were found to have plaque associated with both primary and permanent teeth. Preventive care should be targeted to improve plaque control thus minimizing the risk of developing periodontal disease as adults. PMID- 11853250 TI - Schinzel-Giedion syndrome: interesting facial and orodental features, and dental management. AB - Schinzel-Giedion syndrome comprises multiple congenital anomalies. The orofacial features include coarse facies, frontal bossing, ocular hypertelorism, anterior open bite and macrodontia. Two cases are presented in which the presence of specific craniofacial anomalies with bilateral hydronephrosis confirmed the diagnosis. In one patient, bottle-feeding was associated with caries in maxillary central and lateral incisors, but the second patient was permanently tube fed and did not experience any dental caries. Clinical management of these patients requires a coordinated approach from a team of medical and dental specialists. PMID- 11853252 TI - Scanning acoustic microscopy study of human cortical and trabecular bone. AB - Scanning acoustic microscopy (SAM) has been used in the burst mode to study the properties of human cortical and trabecular bone. An Olympus UH3 SAM (Olympus Co., Tokyo, Japan) was used with a 400 MHz burst mode lens (120 degrees aperture, nominal lateral resolution 2.5 microm). The human cortical bone was from the midshaft of a femur from a 60+ male cadaver; the trabecular bone specimens were obtained from the distal femoral condyles of another 60+ human male cadaver. Elastic moduli for both trabecular and cortical bone were obtained by means of a series of calibration curves correlating SAM gray levels of known materials with their elastic moduli; specimens included: polypropylene, PMMA, Teflon, aluminum, Pyrex glass, titanium, and stainless steel. Values obtained by this method are in good agreement with those obtained by nanoindentation techniques. The three critical findings earlier by Katz and Meunier were observed here as well in both the cortical and trabecular bone samples. PMID- 11853253 TI - Quantitative measurements of the mechanical properties of human bone tissues by scanning acoustic microscopy. AB - The quantitative scanning acoustic microscopy is used for determination of the acoustic impedance of human femoral bones in the micrometer scale. The method is introduced and demonstrated at prepared bone surfaces with different cutting angles referring to the bone axis. First measurements show weak variations of the elastomechanical properties with angles. PMID- 11853254 TI - Application of scanning acoustic microscopy for assessing stress distribution in atherosclerotic plaque. AB - Scanning acoustic microscopy (SAM) was equipped to assess the acoustic properties of normal and atherosclerotic coronary arteries. The SAM image in the atherosclerotic lesion clearly demonstrated that the sound speed was higher than that in the normal intima, and that the variation of elasticity was found within the fibrous cap of the plaque. Young's elastic modulus of each region was calculated and the finite element analysis was applied to derive the stress distribution in these arterial walls. In a case of normal coronary artery, the stress was dominant in the intima and the distribution was rather homogeneous and in a case of atherosclerosis, high stress was concentrated to the relatively soft lesion in the fibrous cap overlying lipid pool. SAM provides information on the physical properties, which cannot be obtained by the optical microscope. The results would help in understanding the pathological features of atherosclerosis. PMID- 11853255 TI - Acoustic microscopic analysis of the biological structure of insect wing membranes with emphasis on their waxy surface. AB - The mechanical performance of natural materials depends on the type, and especially the composition of the molecular constituents. They are almost without exception composite materials, whose characteristics are determined by the characteristics of the individual constituents, their shape, their interaction, and in particular their orientation within the natural material. One of the most impressive natural composites is the insect cuticle. This lightweight building material impresses one with its ability to withstand extremely heavy loading. Even the ultrathin (3-10 microm) membranes of insect wings add greatly to the structural stability of the wings. By means of acoustic microscopy, the present study also shows that the thin covering of wax on the membrane is not an accidental material arrangement. Contrary to that of locust wings, dragonfly wing membranes were found to have a criss-cross fiber-like density gradient within to the waxy layer. This density gradient proved to be mechanically relevant in stabilizing the wings. PMID- 11853256 TI - Small intestine wall distribution of elastic stiffness measured with 500 MHz scanning acoustic microscopy. AB - Obtaining data relating intestinal mechanical properties and histology is a step towards the next level in the hierarchy of structure of living tissue, and may provide new insight into the mechanisms of intestinal function and disease such as obstruction. Due to lack of methodology, however, such data are currently sparse. Scanning acoustic microscopy (SAM) can measure the propagation speed of sound (C) and the acoustic impedance (Z) with micrometer resolution in tissue. By use of elementary theory of elasticity, the elastic stiffness (c11) can be computed from C and Z. We used 5-microm-thick transverse sections of ethanol treated guinea pig small intestine as the experimental model and measured the distribution of C and Z across the intestinal wall using SAM at 500 MHz. The individual layers mucosa, submucosa, and circular and longitudinal muscle were discerned with ease in the images and varied significantly with respect to both C and Z in most cases. The measured values (median values) of C ranged from 1550 to 1669 m s(-1), and Z ranged from 2.10 to 2.60 MPa s m(-1). c11 differed between all layers ranging from 3.25 to 4.27 GPa with the following sequence of magnitude: circular muscle >submucosa>mucosa>longitudinal muscle (p<0.001). In conclusion, we provided the first microscale mechanical data relating to the histological layers of the small intestine. PMID- 11853257 TI - Structural and radiological parameters for the nondestructive characterization of trabecular bone. AB - Trabecular bone is characterized by compositional and organizational factors. The former include porosity at microlevel and mineralization. The latter refer to the trabecular architecture. Both determine the mechanical properties of the trabecular bone. The aim of this study is to investigate the relationship between the mechanical properties and the local HU value, the bone mineral density, the in vitro histomorphometric properties assessed by means of microcomputed tomography, and the Young's modulus determined by ultrasound measurement. Also the correlation between local HU values based on CT data of the full bone and HU values based on CT data of excised trabecular bone cylinders is investigated. Therefore density and strength related parameters of 22 trabecular bone cylinders retrieved from a fresh cadaver femur were measured by using different techniques. The mean HU value of the excised bone samples is very highly correlated with the pQCT density (R2=0.95) and the microCT-based morphometric parameter BV/TV (R2=0.95). The mean HU values, determined from the CT images of the planned and excised bone samples, are less highly correlated (R2=0.75). The Young's modulus E(US) determined from the ultrasound measurement is highly correlated with the maximal stress sigmamax (R2 = 0.88) but not with the mechanically determined Young's modulus Emech (R2 = 0.67). The maximal stress sigmamax correlates well with the density parameters (R2 varies between 0.76 and 0.86). On the contrary the mechanically determined Young's modulus Emech does not correlate well with the density parameters (R2 varies between 0.52 and 0.56). The absorbed energy Eabs during the deformation is only highly correlated with the maximal stress sigmamax (R2 = 0.83). The inclusion of structural parameters besides a density related parameter did improve the prediction of the Young's modulus and the maximal stress. In conclusion, it seems that the HU value from clinical CT scanning is a good predictor of the local bone density and volume fraction. A combination of local density and a measure of the structural anisotropy is clearly needed to achieve good predictions of bone mechanics. PMID- 11853258 TI - Investigation of the morphology of the lacunocanalicular system of cortical bone using atomic force microscopy. AB - Mechanical loading has been implicated as a powerful driving mechanism for interstitial fluid flow through bone. However, little information is available with regard to the morphology of bone fluid spaces, e.g., the canalicular wall, which would be expected to dictate the type of flow regime developing in the lacunocanalicular system under mechanical loads. The purpose of this study was to examine the fine structure of the lacunocanalicular system in cortical bone using atomic force microscopy (AFM), resin casting methods, and selective etching of the specimen surface. A resin-cast replica of the canalicular wall was produced and surface morphology and dimensions were observed using AFM in tapping mode. Material contrast was obtained using surface potential measurements. A striped pattern perpendicular to the canaliculus long axis with a periodicity of 125 nm dominated the structure of the canalicular wall; it is likely that this was caused by the imprint of collagen fibrils arranged in parallel, lining the canaliculus wall. The largest dimension measured for canalicular diameter was on the order of 500 nm. The regular dips and ridges caused by the collagen that lines the wall are a source of roughness which may influence shear stresses imparted by the fluid on the cell surface as well as mixing of solutes within the lacunocanalicular system. In addition, the lacunocanalicular wall lining is likely to affect physicochemical interactions between the fluid and bone matrix. This has important implications for modeling and understanding the microfluid mechanics and rheology of the fluid-filled lacunocanalicular network. PMID- 11853259 TI - Relationship between ultrastructure and the nanoindentation properties of intramuscular herring bones. AB - The nanoindentation technique was used to characterize the indentation modulus of intramuscular herring bone (Clupea harengus) along its length, from early to fully mineralized regions. Both the indentation modulus and modulus anisotropy ratio, indentation modulus longitudinally/indentation modulus transversely, increased nonmonotonically with mineralization, from 1.1 in the early stages of mineralization to 2.1 in the fully mineralized region. The indentation modulus in the fully mineralized region decreased from 19.9+/-2.0 GPa in the longitudinal direction to 11.4+/-1.5 GPa in the transverse directions. In the earliest stages of mineralization, the indentation modulus was 3.8+/-0.7 GPa in the longitudinal direction and 3.5+/-0.3 GPa in the transverse direction. High-intensity synchrotron x rays were used to examine two parameters of crystal texture, the coherence length and the angular spread. Low angular spread was seen along the c axis of bone crystallite. However, the large angular spread observed along the (300) plane indicated an imperfection of crystallite periodicity in the direction perpendicular to the longitudinal axis in the bone. The relatively high coherence length was related to the low angular spread. In general, the indentation modulus increased with increasing crystallite size, but the indentation modulus decreased with increasing crystallite size along the (300) plane. PMID- 11853260 TI - Strength and electron features of irradiated bone. AB - Radiation applied for therapy and diagnostics damages molecular/atomic couples of biotissues, bones being among them. As a result, electron peculiarities and mechanical behavior of the latter are altered. Correlation between strength of the irradiated bone and its electron features explored due to photo- and exoelectron emission measurements is reviewed. PMID- 11853261 TI - Scanning force microscopy observation of tumor cells treated with hematoporphyrin IX derivatives. AB - Cells from human head-neck squamous cell carcinoma (hypopharynx, UMSCC11, and UMSCC22) and lung tumor (alveolus, UMSCC7) were investigated by light and scanning force microscopy (SFM) observation. In the present study a less-invasive contact mode was used to scan living cells in air covered with a thin fluid layer. The investigations were done without any pretreatment of the specimens for such observations as chemical fixation, or staining. Untreated tumor cells, and those treated with the antitumor drug hematoporphyrin IX derivative (HpD) were studied without photosensitizing. Additionally, the temperature influence on cell proliferation was studied. Three-dimensional topographic images and their magnifications offer highly informative insights into the untreated cell surface. In the present study, the cell structure destruction and cell death could partly be visualized by observing the head-neck tumor cells incubated with HpD. Most of the drug-treated head-neck tumor cells died (only 2%-5% of survivors). However, HpD could not affect cells of the lung tumor cell type. They, well known as more resistant against oxidation processes, survived to a great extent. Also, no distortion of the membrane under drug treatment could be observed. The possibilities and limits in the use of SFM for studying the topography of cell surfaces are presented in many details. PMID- 11853262 TI - Blood cell adhesion on sensor materials studied by light, scanning electron, and atomic-force microscopy. AB - Unwanted interactions of biomedical sensors with surrounding tissues, body fluids, and cells are one of the most crucial problems affecting their long-term stability. In vivo processes were simulated in a computer-controlled bioreactor connected to a flow chamber system. Optical sensor materials were inserted into a parallel-plate chamber and monitored by light microscopy in order to get information about the number of adhered cells. Tests with thrombocyte-enriched plasma show that novel phosphorylcholine (PC)-polymer-coated sensors appear to be more bioinert, and thus demonstrate better haemocopatibility in comparison with untreated glass sensors. The influence of different materials on the morphology of adhered cells was investigated by off-line methods such as scanning electron microscopy (SEM) and atomic-force microscopy (AFM). SEM showed a reduction in the number of adhered thrombocytes and the lack of any fibrin network on the PC polymer-modified glass surface, in contrast to the pure glass surface. AFM gives topographical information, and the calculated contact areas and cell volumes indicate smaller interactions between cells and sensor material in the case of PC polymer-coated sensors. PMID- 11853263 TI - Noninvasive measurement of acceleration at the knee joint in patients with rheumatoid arthritis and spondyloarthropathy of the knee. AB - Spondyloarthropathy represents a group of joint diseases with a tendency to reactive new bone formation. Spondyloarthropathy includes Reiter's syndrome, ankylosing spondylitis, and the arthropathy of inflammatory diseases (ulcerative colitis and Crohn's disease). Usually, an extensive investigation is required to distinguish spondyloarthropathy of the knee joint from rheumatoid arthritis. Recently, Reddy et al. (Ann. Biomed. Eng. 23:78-84, 1995) have developed the accelerometry technique to characterize various types of arthritis. The question remains if noninvasive acceleration measurements can be used to distinguish between spondyloarthropathy and rheumatoid arthritis. An ultraminiature accelerometer was placed on the patella, and the subject was asked to rhythmically rotate the knee from 90 flexion to full extension. Results have shown that the mean power of acceleration signal in the range of 100-500 Hz is significantly different (p<0.05) for spondyloarthropathy patients when compared to rheumatoid arthritis patients. The noninvasive accelerometry technique represents a potential tool for characterization of spondyloarthropathy patients. PMID- 11853264 TI - Determination of an optimal kinematic protocol for computer-assisted evaluation of anterior cruciate ligament deficiency. AB - The recent development of computer-assisted techniques for surgery, anterior cruciate ligament (ACL) reconstruction in particular, provides new ways of improving intraoperative kinematic evaluation and final functional outcome. In this paper we have tried to identify the optimal protocol for ACL evaluation, which can fully exploit the novel capability of electronic sensors and computer elaboration. In this work we statistically compared the main clinical tests that are used for knee evaluation and three different numerical methods for kinematic analysis. Results showed that only passive tests that investigate rotational and anteroposterior stability could discriminate ACL status and that the most effective protocol for computerized evaluation should be based on the Lachman and Drawer test, and forced internal-external rotations at full extension for pigs and 60 degrees, elaborated with the functional method (Martelli et al. Comput. Methods Programs Biomed. 62:77-86, 2000). PMID- 11853265 TI - Effect of muscle biomechanics on the quantification of spasticity. AB - The impact of muscle biomechanics on spasticity was assessed by comparison of the reflex responses of the elbow and metacarpophalangeal (MCP) flexor muscles in individuals with chronic spastic hemiplegia following stroke. Specifically, methods were developed to quantify reflex responses and to normalize these responses for comparison across different muscle groups. Stretch reflexes were elicited in the muscles of interest by constant velocity ramp-and-hold stretches at the corresponding joint. The muscles were initially passive, with the joint placed in a midrange position. Estimates of biomechanical parameters were used to convert measured reflex joint torque and joint angle into composite flexor muscle stress and stretch. We found that the stretch reflex response for the MCP muscle group had a 74% greater mean stiffness modulus than that for the elbow muscle group, and that the reflex threshold was initiated at an 80% shorter mean muscle stretch. However, we determined that initial normalized fiber length was significantly greater for the experiments involving the MCP muscles than for those involving the elbow muscles. Increasing the initial composite fiber length of the elbow flexors produced significant reduction of the reflex threshold (p<0.001), while decreasing the initial length of the MCP flexors significantly reduced their measured reflex stiffness (p<0.001). Thus, biomechanical parameters of muscle do appear to have an important effect on the stretch reflex in individuals with impairment following stroke, and this effect should be accounted for when attempting to quantify spasticity. PMID- 11853267 TI - Available space and extracellular transport of macromolecules: effects of pore size and connectedness. AB - Molecular exclusion in tumor tissues is one of the limiting factors for drug delivery to tumor cells. It can be quantified by the available volume fraction of solutes (K(AV)). We found in a previous study that K(AV) of dextran in tumor tissues decreased sharply when the molecular weight (MW) of dextran was increased from 40,000 to 70,000. Outside this range, K(AV) was less sensitive to the MW of dextran. To understand the mechanisms of the MW dependence of K(AV), we investigated K(AV) in tissue phantoms composed of tumor cells in 1% agarose gels, and performed numerical simulations of the available volume fraction in pore networks. We found that the MW dependence of K(AV) in tissue phantoms was similar to that in tumor tissues when the volume fraction of cells in the former was approximately 30%. Our numerical simulations revealed that the sharp decrease in K(AV) required two necessary conditions: (i) the existence of at least two populations of pores and (ii) the lack of connectedness of available pores in the interstitial space. Furthermore, results in this study suggest that it is important to consider not only the local structures of pores but also their connectedness in analyses of molecular transport in tissues. PMID- 11853266 TI - General strategy for hierarchical decomposition of multivariate time series: implications for temporal lobe seizures. AB - We describe a novel method for the analysis of multivariate time series that exploits the dynamic relationships among the multiple signals. The approach resolves the multivariate time series into hierarchically dependent underlying sources, each driven by noise input and influencing subordinate sources in the hierarchy. Implementation of this hierarchical decomposition (HD) combines principal components analysis (PCA), autoregressive modeling, and a novel search strategy among orthogonal rotations. For model systems conforming to this hierarchical structure, HD accurately extracts the underlying sources, whereas PCA or independent components analysis does not. The interdependencies of cortical, subcortical, and brainstem networks suggest application of HD to multivariate measures of brain activity. We show first that HD indeed resolves temporal lobe ictal electrocorticographic data into nearly hierarchical form. A previous analysis of these data identified characteristic nonlinearities in the PCA-derived temporal components that resembled those seen in absence (petit mal) seizure electroencephalographic traces. However, the components containing these characteristic nonlinearities accounted for only a small fraction of the power. Analysis of these data with HD reveals furthermore that components containing characteristic nonlinearities, though small, can be at the origin of the hierarchy. This finding supports the link between temporal lobe and absence epilepsy. PMID- 11853268 TI - Correction of severe enophthalmos with titanium mesh. AB - The aim of this study was to analyse the types of facial fractures that caused severe enophthalmos and the outcome after repair of the medial wall and the floor of the orbit with titanium mesh. Eighteen cases with severe enophthalmos were repaired with titanium mesh in Siriraj Hospital between January 1992 and December 1999. We recorded the types of facial fractures of the patients, the results after operations and the complications. All cases were followed-up for at least 6 months. Le Fort fracture was the most common cause of severe enophthalmos from orbital fracture. The results after operations were satisfactory in 17 cases. Only one case still had enophthalmos after the first correction with titanium mesh and needed further repair with bone graft. Few complications happened post operatively. They were 4 cases of chemosis and 4 cases of temporary diplopia. All subsided spontaneously. The author concludes that titanium mesh can be used to repair the medial wall and the floor of the orbit with good results. PMID- 11853269 TI - Diplopia in facial fractures. AB - A prospective study was made on facial fracture patients who received treatment in Siriraj Hospital from 1 Jan 1997 to 31 Dec 1998. We recorded the types of fractures which involved the orbit, diplopia, operations and results. From the 675 facial fracture patients in this period, there were 256 cases that involved the orbit. Diplopia was found in 17 cases; Orbital floor fracture 8, Zygomatic fracture 4, Le Fort fracture 4, and Medial aspect fracture of maxilla 1. Ten cases were recent fractures and seven cases were old fractures. The floor of the orbits were explored in all cases and repaired with silastic sheath, except 2 severe enophthalmos cases whose medial wall and the floor of the orbits were repaired with titanium mesh. Diplopia disappeared after operation in 14 cases. Three cases still had minimal diplopia in the upward-gaze position and all of them were old fracture cases. The authors concluded that diplopia was found in 6.64 per cent of orbital fractures. Fracture of the floor of the orbit was the most common cause. An early operation will give a better result than late repair. PMID- 11853270 TI - Effect of sacral buttress technique in Ramathibodi spinal system fixation on L5 S1 motion. AB - This is a retrospective study which reviewed the effects of the sacral buttress technique of Ramathibodi Spinal System (RSS) instrumentation for degenerative conditions of the spine on L5-S1 motion. The inclusion criteria included patients who were treated by wide decompression and fusion of the lumbar spine. Two treatment groups of a different fusion technique were identified; the first group was treated by conventional fixation and the second group was treated by the sacral buttress technique. The objective of this study was to evaluate the effect on L5-S1 motion and translation between the conventional group and sacral buttress group. The material consisted of 66 patients who were operated from 1996 to 1999 for degenerative conditions of the lumbosacral spine. The conventional group included 25 patients in whom instrumentation was stopped at the fifth lumbar spine and the buttress group consisted of 41 patients who underwent the sacral buttress technique with RSS fixation. Dynamic lateral radiographs were measured to determine the motion and translation before and after surgery. In the group with sacral buttress technique, post operation decrease in motion of the lumbosacral joint was statistically significant. Reduced motion was also associated with the level of fusion. The use of the sacral buttress technique with RSS led to significant control in lumbosacral motion without the need for screw fixation to the sacral spine. The usefullness of this technique is a valuable adjunct to spinal fusion in patients who undergo decompressed spinal surgery and may prevent the junctional problem of fusion at this segment and also the upper end of the above fusion level. PMID- 11853271 TI - Early recovery after retropleural approach in a paraparesis thoracic disc patient. AB - A 56 years old Thai male developed acute paraparesis 2 days before admission. The investigation showed T6-7 thoracic disc herniation on the left side with thoracic cord compression. A lateral retropleural (extracavity) approach from the left side without penetrating the pleural cavity was performed. He was able to sit up and start the rehabilitation program within the first day after the operation on the bed and neurological status was gradually recovered within 2 months after continue home program for rehabilitation. Finally he can walk with gait aids. The motor power of his legs were grade IV according to Frankel classification and he can well control the uro-genital function. PMID- 11853272 TI - Limb measurements for height and bone mineral density estimation. AB - To estimate height and forearm bone mineral density (BMD), three simple sites of limb segments of 428 healthy subjects (321 females and 107 males) were measured. Mean length of upper arm length (UAL), lower arm length (LAL) and knee to floor height (KFH) were not different in various age groups (p > 0.05). Good correlations were obtained from all measurements compared with height (r = 0.789, 0.826 and 0.810 for UAL, LAL and KFH, respectively) and moderate correlations were found compared with BMD (r = 0.386, 0.461 and 0.458 for UAL, LAL and KFH, respectively). If physically disabled activity occurred in some segmental limbs, the others which have good ability can be used for height assessment. When applying the stepwise method for multiple regression analysis, combination of LAL, KFH and UAL showed the best correlation (r = 0.866) with height. It indicated that UAL, LAL and KFH measurements provide enough predictive ability for height estimation. Only LAL and KFH were the variables which predicted BMD equation with moderately predictive ability. In primary health care, simple long bone measurements with the patient's history should be a primary step for screening of high risk osteoporotic patients. In prevention of osteoporotic fracture, LAL and KFH may be a useful adjunct to other criteria in the discrimination of high risk patients. PMID- 11853273 TI - Arm span, height and forearm bone mineral density in normal young and postmenopausal women. AB - The value of arm span and height measurements in the erect position were evaluated by (1) comparing them in individual young adults and postmenopausal women and (2) comparing them with forearm bone mineral density (BMD) using a Panasonic dual energy X-ray absorptiometry (DXA). The study group consisted of 301 healthy women composed of 165 young adults (20-40 years), 62 postmenopausal women and 74 women whose age was more than 40 and had regularity of menstruation. Arm span and height of the individual young adults and postmenopausal women were not significantly different (p>0.05). It indicated that arm span and height measurements were independent of normal aging. Height loss occurred in some elderly women who had some diseases or conditions which induced short stature. We suggested that arm span can be used to predict height during the younger adult life of elderly women and to predict the current height in patients who had a height loss problem. A simple linear regression equation of height and arm span has already been constructed. During the increase of years since menopause in postmenopausal women, height and arm span measurements were relatively constant, while BMD gradually declined. We also found that the greater the number of years since menopause, the higher the rate of BMD loss (%). Height and arm span had low correlation with BMD. Height or arm span alone can not be used to predict BMD, but may be combined with other risk factors of osteoporosis. PMID- 11853274 TI - Transient osteoporosis with bilateral fracture of the neck of the femur during pregnancy: a case report. AB - A case of bilateral non-nion of femoral neck fracture during the last trimester of pregnancy was reported. Bilateral hemiarthroplasty with bipolar prosthesis was performed in May, 2000. The result after one year follow-up was excellent. PMID- 11853275 TI - Interspinous bursa and spinal instability. AB - STUDY DESIGN: This is a prospective study. Interspinous tissue was taken intraoperatively for pathological examination for the presence of bursa. The pathologist was unaware of the X-ray findings in each specimen. The presence of bursa was then correlated with X-ray evidence of hypermobility in each segment. OBJECTIVE: To verify the pathogenesis of interspinous bursal formation. SUMMARY OF BACKGROUND DATA: Interspinous bursa is common in the older population. It has been associated with degenerative lumbar diseases, aging and anatomical distance between the spinous process. However, no detailed exploration of the segmental instability as a cause of bursal formation has been done. METHOD: The insterspinous tissue was taken intraoperatively from patients diagnosed as multilevelled spinal stenosis who underwent extensive decompression, fusion and instrumentation. The specimens were examined by the same pathologist for the existence of bursa. The presence of bursa was correlated with X-ray motion study of each spinal segment by student t-test. RESULTS: The existence of bursal was significantly correlated with angular mobility of more than 10 degrees. CONCLUSION: Angular mobility is a possible cause of interspinous bursa. On the contrary, the presence of insterspinous bursa may be evidence of segmental hypermobility. PMID- 11853276 TI - The efficacy of an aerobic exercise and health education program for treatment of chronic low back pain. AB - Low back pain is one of the most common and important musculoskeletal disorders. In addition, chronic low back pain can deteriorate the patient's physical, psychosocial and socioeconomic status. The objective of this quasi-experimental research was to assess the efficacy of an aerobic exercise and health education program in the treatment of chronic low back pain. Seventy-two patients whose ages ranged from 30 to 50 years who had chronic low back pain were enrolled and randomly assigned into two groups. Eight men and 28 women in the experimental group participated in a series of 3 health education sessions and an aerobic exercise training program. Nine men and 27 women in the control group received regular health education and a lumbar flexion exercise program. After a 3-month period of treatment, the results revealed the experimental group had statistically significant improvement of pain score and resting pulse rates when compared to the values of the control group (p-value < 0.001 and < 0.01, respectively). The average serum High Density Lipoprotein-Cholesterol (HDL-C) in the experimental group was also significantly higher (p-value < 0.05) than that of the control group. This health education program is useful and may be applicable to patients with chronic low back pain as an alternative treatment. PMID- 11853277 TI - Prevalence of hypovitaminosis D in elderly women living in urban area of Khon Kaen province, Thailand. AB - This study included 106 volunteer elderly women who were living in an urban area of Khon Kaen province. The mean (+/-SD) of age and of serum 25(OH)D concentration of these elderly women was 69.42 (+/-6.77) years and 33.32 (+/-7.14) ng/ml respectively. There was a significant inverse relationship between serum 25(OH)D and PTH concentration. Serum PTH concentration started to increase steeply when serum 25(OH)D concentration declined < or = 35 ng/ml and increased significantly when serum 25(OH)D concentration declined < or = 30 ng/ml. Furthermore mean (+/ SD) of serum 25(OH)D concentration in the group of osteopenia and osteoporosis of femoral neck was 35.25 (+/-6.77) and 30.92 (+/-6.49) ng/ml respectively. As a result, the prevalence of hypovitaminosis D in these selected elderly women would be at least 34.9 per cent, or as high as 65.1 per cent. PMID- 11853278 TI - Safety and efficacy of meloxicam 7.5 mg in the treatment of osteoarthritis in Thai patients. AB - A prospective, open trial to evaluate the safety and efficacy of oral meloxicam 7.5 mg once daily over a period of 28 days in Thai patients with osteoarthritis was conducted in 3 major hospitals in Bangkok, Thailand. A total of 137 patients were enrolled and completed the study protocol. The mean age of the patients was 57.6 years and 88 per cent were female. Pain on movement and pain at rest evaluated after treatment by visual analog scale (VAS) were significantly improved with respect to the baseline status (p<0.001). The final efficacy was reported as satisfactory or good in 97 per cent and 94 per cent as determined by patients and physicians respectively. Patient status evaluated at the end of the study reported improvement of their arthritic condition in 84 per cent. Drug tolerability assessed by patients and physicians and was good or satisfactory in 99 per cent and 98 per cent. GI adverse event was reported in 8.8 per cent. Other adverse events were itching/rash in 2.0 per cent and headache/insomnia in 1.5 per cent of patients. No patients withdrew from the study due to adverse events. No GI bleeding or perforation were observed. No hospitalization was observed during the study. There was no significant change of blood chemistry and hematological profile between pre and post treatment examination. Results from this study suggest that oral meloxicam 7.5 mg for 28 days is a safe and effective treatment regimen with high GI tolerability profile for the treatment of osteoarthritis in Thai patients. PMID- 11853279 TI - Analysis of polymorphism of the interleukin-6 gene in Thai subjects with osteoporosis. AB - Osteoporosis is the imbalance between bone formation by osteoblast and bone resorption by osteoclast. The genetic factors play an important role in determining bone mass and several genes probably act as regulators of this process. Interkeukin-6 (IL-6) is one of the candidate genes to regulate bone density, since IL-6 has some effect on stimulation of osteoclast resorption and has been implicated in the pathogenesis of bone loss associated with estrogen deficiency. We investigated the relationship between bone mineral density (BMD) and a polymorphic AT rich repeat in the 3' flank of the IL-6 gene in 272 Thai subjects. The subjects were classified into 3 groups i.e. normal healthy control (n=95), border-line (n=112) and osteoporotic patients (n=65). Five alleles different in sizes were identified (designated a, b, c, e and f). It was observed that c/c was the most common genotype in Thais (86.76%). The other genotype frequencies were 0.74, 3.31, 8.09, 0.74 and 0.37 for a/c, b/c, c/e, c/f and b/e genotypes, respectively. The common genotype was different from the Caucasians in a previous study. These frequencies were significantly different from the Caucasians (p<0.05). There was no significant relationship between 3' flanking AT repeat of the IL-6 genotypes and the BMD values of the distal forearm that were determined by One-way ANOVA (p>0.05). Additionally, the impact of the IL-6 genotypes on risk of osteoporosis was assessed by determination of the odds ratio. The c/e genotype may be a protective factor of osteoporosis. On the contrary, the b/c and c/c genotypes were considered to be risk factors of osteoporosis. PMID- 11853280 TI - Efficacy and safety of Menatetrenone-4 postmenopausal Thai women. AB - Clinical and comparative study of the efficacy and adverse events of Menatetrenone-4. The control group (n=40) received elemental calcium carbonate 800 mg/day and the Menatetrenone-4 treated group received elemental calcium carbonate 800 mg/day plus vitamin K2 45 mg/day (n=43). The vitamin K2 treated group showed a marked decrease of undercarboxylated osteocalcin at 2 weeks, six months (51.52% p=0.0001) and twelve months (87.26% p=0.0001) compared to the calcium treated group. At the end of the sixth and twelve months both groups did not increase bone mass of the hip but the vitamin K2 treated group increased 0.6 per cent of bone mass of the lumbar spine and decreased bone resorption 65.42 per cent (p=0.0001) compared to the calcium treated group. The calcium treated group was switched to the vitamin K2 treated group at the end of six months and showed a decrease of the level of undercarboxylated osteocalcin the same as the former vitamin K2 treated group. The adverse events were 2 cases of mild skin rash which subsided after cessation of medication. PMID- 11853281 TI - Study of the level of biochemical bone markers: NMID osteocalcin and bone resorptive marker (beta CTx) in Thai women. AB - Study of the level of NMID osteocalcin and Beta CTx in 700 Thai women. The mean of NMID osteocalcin = 21.45 SD = 11.18 95% CI = 20.12 to 22.79 ng/ml and betacrosslap = 0.445 ng/ml SD = 0.25 with 95% CI = 0.414 to 0.478 ng/ml in women who menopaused less than 10 years ago (n=237), these values were higher than in young adult females (n=63) and menstruating women (n=123) which was statistically significant (p=0.0001). The value of both bone markers in elderly women who had menopaused more than 10 years ago and were aged more than 60 years (n=94) showed a marked increase of NMID osteocalcin, 25.63 ng/ml SD = 14.22 ng/ml but the value of betacrosslap was below the young-menopausal women, 0.394 ng/ml SD = 0.241 ng/ml 95% CI = 0.344 to 0.444 ng/ml. Menopausal women are at a high risk of osteoporosis due to high bone turnover. In our study, the NMID osteocalcin had a high correlation with betacrosslap (r=0.789 p=0.0001) while both bone markers had a weak correlation with bone mass density of radius, lumbar spine and hip by DXA. (r=0.29 p=0.0001). PMID- 11853282 TI - The parathyroid hormone level in Thai women. AB - Study of the correlation of parathyroid hormone (PTH) with age, radial bone mass and Nitrogenous mid fragment osteocalcin (NMID osteocalcin) in 226 menopausal women and 123 menstrual women. In menopausal women, aged between 50 and 69, the level of PTH did not increase with age (r=0.001, p=NS). Elderly women (n=123, age>70) showed a slight increase of PTH, 7.8 per cent compared to menstruating women. In elderly women (n=100, age>60) there was no weak correlation (r=0.11, p=0.0001) with bone mass of the distal end of the radius measured by Dual X-ray Absorptiometry (DXA) (Panasonic BDM-3) and no correlation between PTH and the resorptive bone marker, (betacrosslap) (r=0.11, n=122). PMID- 11853283 TI - Serum dehydroepiandrosterone sulfate, testosterone, and biochemical markers of bone turnover in elderly Thai men. AB - The most abundant human steroid, dehydroepiandrosterone sulfate (DHEAS), may have a multitude of beneficial effects, but declines with age. It is unclear whether DHEAS deficiency is an important factor contributing to increased bone resorption and impaired bone formation or not that leads to their bone loss. Thus, we investigated serum DHEAS, testosterone, osteocalcin (N-MID osteocalcin) and C terminal telopeptides (beta-CrossLaps) in 121 healthy Thai males without bone diseases. Thirty-nine males (mean age 31.5 +/- 8.2, range 23-42 years) were recruited as the normal adult group and 82 males (mean age 61.2 +/- 7.0, range 52 77 years) were assigned as the elderly group. DHEAS levels were higher in the adult group compared with the elderly subjects (296.8 +/- 93.4 vs 172.6 +/- 99.8 microg/dL, p < 0.0001). Serum osteocalcin concentrations were also higher in the adult group compared with the elderly males (27.9 +/- 11.1 vs 23.2 +/- 7.9 ng/ml, p = 0.0091). However, serum testosterone and C-terminal telopeptides levels were not significantly different between the two groups. We concluded that low DHEAS concentrations are commonly encountered in elderly males and may relate to low osteocalcin levels due to the osteoblast stimulation effects of DHEAS. These findings may be implicated in the treatment of osteoporosis in elderly men by using DHEAS. PMID- 11853284 TI - Treatment of painful osteoarthritis of the knee with hyaluronic acid. Results of a multicenter Asian study. AB - Hyaluronic acid is a component of synovial fluid, in osteoarthritis (OA) and shows a change of property of hyaluronic acid especially its viscoelasticity causing degradation of the cartilage. Injection of hyaluronic acid into OA of the knee (Hyalgan 2 ml/wk for 4 doses) in international multicentres gave dramatic responses: decreased pain, increased mobility of the joint after 49 days of injections compared to a placebo (2 ml of normal saline) p<0.01. PMID- 11853285 TI - Efficacy of calcitriol to bone resorption and PTH level: clinical study. AB - Study of the effect of calcitriol (Rocaltrol) to bone markers and PTH level in 146 menopausal women. The result showed a dose of calcitriol 0.75 microgram/day for one month significantly decreased the level of PTH, bone marker for resorption and NMID osteocalcin 61.55, 69.92, 60.02 per cent respectively (p=0.000). The other group which took calcitriol 0.5 microgram/day had little change to such markers. The active form of vitamin D benefits bone health by enhancing osteocalcin formation, in addition vitamin D has moderate antiresorption and suppression of PTH secretion. PMID- 11853286 TI - Difference in bone acquisition among hormonally treated postmenopausal women with normal and low bone mass. AB - This prospective analysis was conducted to compare the effects of hormone treatments on bone mineral density (BMD) of the lumbar spine, hip, and distal forearm in postmenopausal women with normal BMD and those with low bone mass. Eighty healthy women were randomly assigned to receive a cyclic regimen of standard hormone replacement therapy (HRT) or currently used low-dose oral contraceptive (OC). Women were categorized as normal BMD and low bone mass according to the Thai reference database. The results revealed that women with low bone mass gained more BMD than those with normal BMD. The difference in mean per cent bone acquisition was obvious at the spine. In addition, further subset analysis into OC and HRT groups revealed higher effects of OC on BMD when compared to HRT. PMID- 11853287 TI - Bone mineral density of lumbar spines and proximal femur in the normal northeastern Thai women. AB - OBJECTIVE: To determine reference data for age-matched bone mineral density (BMD) of normal northeastern Thai women. MATERIAL AND METHOD: 350 northeastern Thai women aged 20-70 years were prospectively studied. After interview, the subjects were examined for BMD of lumbar spines and proximal femur. Mean BMD of each group was determined for each skeletal site. Standard deviation (SD) of BMD in the peak group was calculated to determine BMD cutoff level for diagnosing osteoporosis. RESULT: Peak average BMD of lumbar spines was at 40-45 years, whereas peak BMD of proximal femur was at 35-40 years. Mean value of peak BMD of lumbar spines and proximal femur was 1.226 and 1.035 g/cm2, respectively. According to T-score less than -2.5 SD criteria, osteoporosis of lumbar spines and proximal femur was considered when measured BMD was below 0.889 and 0.785 g/cm2, respectively. CONCLUSION: Our study provided the BMD reference data for northeastern Thai women. PMID- 11853288 TI - Diagnostic agreement of combined radiogrammetric analysis with texture analysis in the evaluation of bone density: a comparison with dual energy X-ray absorptiometry. AB - To assess the reliability and validity of combined radiogrammetric analysis with texture analysis (digital X-ray radiogrammetry: DXR) of metacarpals and distal forearm when compared to dual energy X-ray absorptiometry (DXA) of the distal radius. DXA readings from the supradistal and distal 1/10 of radius were compared with DXR obtained on the same day with a sample of 274 women and 87 men (average age 31.33 years, range 20-65 years). The results showed that both sites of forearm DXA scores were significantly correlated with DXR scores (r = 0.5211 and 0.6090, respectively). DXA scores were used as the standard, defining moderate fracture risk by a standardized t score <-2 and marked risk by t <-3. Following the current WHO definition of osteoporosis, t <-2.5 was also applied. Using cutting points of t <-2, <-2.5 and <-3 for DXA and t <-2 for DXR values, the sensitivity to osteopenia was 38.98 per cent, 40.63 per cent and 55.56 per cent respectively, with specificity of 94.36 per cent for all cutting points of DXA. At these cutting points, the corresponding false positive was 5.64 per cent and false negatives were 61.02 per cent, 59.37 per cent and 44.44 per cent, respectivety. It indicated that DXR measurement had low sensitivity but was appropriate with high specificity for discrimination of forearm osteoporosis. PMID- 11853289 TI - DHEA(S): the fountain of youth. AB - Dehydroepiandrosterone (DHEA) and its sulfate ester (DHEAS) are weak androgens produced primarily by the adrenal gland. Although their plasma concentrations by far exceed those of any other adrenal product, their physiological roles have not yet been determined. In plasma, where the major portion of these hormones is present in the sulfate form, it is possible that DHEAS serves as a reservoir for DHEA. Since various tissues have been shown to contain steroid sulfatases. The peak plasma levels of DHEA and DHEAS occur at approximately age 25 years, decrease progressively thereafter, and diminish by 95 per cent around the age of 85 years. The decline of DHEAS concentrations with aging has led to the suggestion that DHEAS could play a role in itself and be implicated in longevity. Moreover, the epidemiological evidence has shown that adult men with high plasma DHEAS levels are less likely to die of cardiovascular disease. DHEA has also been shown to increase the body's ability to transform food into energy and burn off excess fat. Another recent finding involves the anti-inflammatory properties of DHEA. It has been known that DHEA can lower the levels of interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-alpha). It should be pointed out that chronic inflammation is known to play a critical role in the development of the killer diseases of aging: heart disease, Alzheimer's disease and certain types of cancer. In conclusion, DHEA or DHEAS administration combined with conventional treatment may be implicated in particular conditions to improve the quality of life. PMID- 11853290 TI - Midpedicular distance: an alternative pedicular distance measurement. AB - OBJECTIVES: 1. To introduce the measurement of midpedicular distance (MPD). 2. To study the correlation of the MPD measured on AP plain film and CT scan film and compared with the correlation of the interpedicular distance (IPD) measurement. MATERIAL AND METHOD: The MPD, the distance between intersection point of midvertical and midhorizontal lines drawn on the two pedicles of the same vertebra, was measured on lumbar spine (L1, L2) AP plain film. The IPD, the distance between the inner cortical shadow of the pedicles, was also measured on the same film. The measurements of MPD and IPD were then carried out on the same individual's CT scan film (which had an imprint scale). The study was performed on 35 subjects (20 males, 15 females) with an average age of 48.2 years (range 19 81 years). The correlation of the value measured on plain film and CT scan film was studied using Pearson's correlation test. RESULT: The MPD measurement on plain film had higher correlation with MPD measurement on CT scan film (L1 level: r = 0.778, L2 level: r = 0.732, p<0.001) compared with the IPD measurement (L1 level: r = 0.672, L2 level: r = 0.604, p<0.001). CONCLUSION: The midpedicular distance measurement on lumbar spine AP plain film can better be used to reflect the pedicular distance than the interpedicular distance. PMID- 11853291 TI - A common-source water-borne outbreak of multidrug-resistant typhoid fever in a rural Thai community. AB - We report an epidemiological investigation of an explosive water-borne outbreak caused by multidrug-resistant (MDR) Salmonella Typhi in a non-endemic community with otherwise good sanitation. Between 31 October and 11 November 1999, 11 cases of typhoid fever occurred in Poppra District, Tak Province. Four cases were children < or = 14 years of age and the remaining 7 were adults 15-24 years old. Hemoculture was performed in 10 of the 11 cases, of whom 5 (50.0%) were positive for Salmonella Typhi. All the isolates were of Vi phage type E14 which was resistant to chloramphenicol, ampicillin, co-trimoxazole, tetracycline, and streptomycin but sensitive to kanamycin, ofloxacin, ciprofloxacin, and cefotaxime. One confirmed case developed intestinal perforation leading to death. A case-control study revealed a significant association between disease and drinking unboiled spring water. The implication of spring water was supported by the presence of faecal contamination in the spring water source. Rapid identification of the possible transmission source and prompt implementation of control measures curtailed the spread of this outbreak. PMID- 11853292 TI - Hemospermia: review of the management with 5 years follow-up. AB - OBJECTIVE: To review our experience with hemospermia and a long term follow-up. MATERIAL AND METHOD: Medical records of patients with hemospermia treated at the Division of Urology, Ramathibodi Hospital between 1993 and 1995 were reviewed. Clinical presentation, investigation, diagnosis, treatment, outcomes and long term follow-up were noted. RESULTS: Sixty-eight patients were found and completed follow-up to 5 years. The mean age was 40 years (range 28-62). Physical examination including DRE and urine examinations were done in all of the cases. Special investigations such as PSA, TRUS, IVP and cystourethroscopy were performed in selected cases. Prostatitis was found in 27.9 per cent, tuberculosis in 4.4 per cent, sexually related causes in 5.8 per cent and idiopathic in 61 per cent No malignancy was found in this study. Hypertension was found in 7.3 per cent of the patients. Thirty-two per cent had recurrent episodes of hemospermia. Specific treatment was used only for prostatitis and tuberculosis. No specific treatment was used for the idiopathic group. CONCLUSIONS: Hemospermia is a benign condition. Most of the causes were from idiopathic and inflammation. Only simple investigation was needed and treatment was recommended depending on the diagnosis and no specific treatment was needed for idiopathic cause. PMID- 11853293 TI - Clinical manifestation of tuberculous and pyogenic spine infection. AB - The authors compared clinical and baseline parameters between patients with tuberculous spondylitis (67 patients) and pyogenic vertebral osteomyelitis (34 patients). The average age was 50.8 (range, 1 to 82 years). Males were slightly more predominant in both pyogenic and tuberculous infections. Seventy per cent of patients with pyogenic infection had had symptoms for less than 3 months, while this was the case for only 44 per cent of the tuberculous patients. Lumbar spine was the most common site of involvement. Associated infections were more commonly found in tuberculous infection. Thirty per cent of tuberculous spondylitic patients were initially misdiagnosed, while 44 per cent of patients with pyogenic vertebral osteomyelitis had delayed diagnosis. Only 20 per cent and 30 per cent of tuberculous and pyogenic patients had fever, respectively. Neurological deficit occurred more frequently in pyogenic than in tuberculous infection (79% vs 59%), while bowel bladder involvement was more commonly found in tuberculous spondylitis. Kyphosis was significantly more common in tuberculosis than in pyogenic infection (50.8% vs 26.5%, respectively). Sinus tract formation, subcutaneous abscess formation and positive for SLRT (straight-leg-raising test) were found in only a small percentage of the patients. Duration of symptoms, site, associated infection, kyphosis and neurological deficit could be used for differentiation of spinal infection. PMID- 11853294 TI - Juvenile dermatomyositis in Thai children. AB - Juvenile dermatomyositis is a rare, chronic multisystemic inflammatory disorder of unknown etiology, characterized by a typical skin rash and proximal muscle weakness. A retrospective study from the medical records of patients diagnosed as juvenile dermatomyositis was performed at Queen Sirikit National Institute of Child Health from 1988 to 1998. There were seven cases of juvenile dermatomyositis diagnosed according to the criteria of Bohan and Peter. Six cases were female and one case was male. The age of diagnosis ranged from 2.5 years to 11 years. (mean age was 7 +/- 3.6 years). The presenting symptoms were muscle weakness (6 cases), muscle pain (2 cases) and skin rashes (4 cases). All of the patients developed proximal muscle weakness of the lower extremities varying from grade 3 to grade 4. The cutaneous manifestations were heliotrope signs (6 cases), gottron's papules (2 cases), photosensitivity (2 cases) and calcinosis cutis (4 cases). Electromyography (EMG) was performed in 6 cases and revealed typical change of myopathic type. Elevated muscle enzymes were noted in all cases. Muscle biopsy was performed in 6 cases and was compatible with myositis. Oral prednisolone (1-2 mg/kg/day) was given in 6 cases and the muscle weakness improved. There was no mortality in this study. Four cases developed calcinosis cutis 1 to 3 years after muscle weakness and did not respond to any treatment. In conclusion, juvenile dermatomyositis is a disease which causes chronic disability in children. Early diagnosis and treatment can prevent morbidity and mortality. Calcification at the skin usually occurs after the onset of muscle weakness several months to years after diagnosis. PMID- 11853295 TI - Transesophageal echocardiography during percutaneous mitral commissurotomy in patients with left atrial thrombus. AB - BACKGROUND: Transesophageal echocardiography (TEE) is used routinely before percutaneous transvenous mitral commissurotomy (PTMC) to detect left atrial appendage thrombus (LAAT) to avoid the risk of embolic complications. The issue of whether patients with small and fixed LAAT should be denied the potential benefit of PTMC is worth examining. OBJECTIVE: To evaluate the safety and efficacy of PTMC with Inoue balloon catheter in mitral stenosis patients with LAAT using TEE continuous monitoring during the procedure. MATERIAL AND METHOD: All TEE studies performed during PTMC and transthoracic echo-cardiography (TTE) performed the same day and repeated on the day after the procedure between March 1995 and January 2000 were reviewed. RESULTS: A total of 1,238 consecutive TEE during PTMC were reviewed. LAAT was detected in 111 patients (mean age 43.7 +/- 10.1 years, male:female = 1:2, atrial fibrillation : sinus rhythm 2.47:1). LAAT were grossly oval with the largest measuring 3.5 x 2.8 centimeters. Mobile LAAT was detected in 3 patients (2.7%), one of whom developed a transient ischemic attack and another had an episode of stroke after PTMC. Mitral valve area (by 2D Echocardiography) pre PTMC was 0.8 +/- 0.2 cm2 and post-PTMC was 1.5 +/- 0.3 cm2. Most of our patients became fully ambulatory and could be discharged from the hospital the day after the procedure, except for two patients who developed severe mitral regurgitation and needed elective mitral valve surgery thereafter. CONCLUSION: PTMC with the Inoue-balloon catheter can be carefully and safely performed in patients with small, fixed LAAT under continuous TEE guidance with acceptable risk. PMID- 11853296 TI - Surgical treatment of Le Fort fractures in Ban Pong Hospital: two decades of experience. AB - A retrospective study of 64 patients with maxillary fracture of Le Fort type who were treated at the Surgical Department, Ban Pong Hospital, Ratchaburi, Thailand during the past 21 years (September 1st, 1979 - August 31st, 2000) is presented. Most of the patients were male (84.4%). Patients mainly affected were in the third decade of life (54.7%) with an age range of 13-65 years old. The etiology of the fracture was mostly related to road traffic accidents (90.6%). The most common type was Le Fort II fracture (54.7%), followed by associated facial bone injury were mandible (47.4%) and associated other organ injuries were fractures elsewhere (50.0%). Open reduction with intermaxillary fixation and maxillary suspension were the treatments of choice and the results were considered to be successful with only mild post-operative complications. PMID- 11853298 TI - Normal plasma free amino acid levels in Thai children. AB - Analysis of plasma free amino acid levels is important for diagnosis of inborn errors of metabolism. Traditionally, this is performed using commercially available dedicated amino acid analyzers, but few such instruments are available in Thailand, and many are not used in routine operations. Here, the authors describe the analysis of plasma free amino acid levels in 57 normal children by reverse-phase HPLC and pre-column derivatization with phenylisothiocyanate. Plasma free amino levels are reported as mean +/- SD and 95 per cent confidence interval of mean for each of 5 age groups: 0-6 months; 6-12 months; 1-3 years; 3 6 years; 6-12 years. Mean amino acid levels were generally similar in all age groups (p > or = 0.01), except that hydroxyproline tended to be higher in the 0-6 months age group compared to other age groups (p<0.01). Comparisons were made between the present data with the normal free plasma amino acid levels in children of similar age groups reported both in Thailand and overseas in terms of both mean +/- SD and maximum and minimum values. Overall, our methodology involving HPLC can identify 35 amino acid derivatives, including all the major amino acids except for cysteine, which is substantially more than the number reported in earlier work on plasma free amino acid levels in normal Thai children. Moreover, the present methodology gives mean +/- SD values similar to an overseas report. For these reasons, HPLC should be considered as an alternative approach in laboratories, where demand does not justify the need for dedicated amino acid analyzers. However, there can be substantial variations between the results from different laboratories, and each laboratory should establish its own normal values. PMID- 11853297 TI - Comparative study of bulky stage IB and IA cervical cancer patients treated by radical hysterectomy with and without neoadjuvant chemotherapy: long-term follow up. AB - One hundred and ninety patients with bulky (> 3 cm) stage IB and IIA cervical cancer who underwent radical hysterectomy between 1991 and 1994 at Maharaj Nakorn Chiang Mai Hospital were reviewed to determine whether neoadjuvant chemotherapy (NAC) with MVAC (Methotrexate, Vinblastine, Adriamycin, Cisplatin) improved survival. There were 42 patients treated with pre-operataive NAC (MVAC 1-3 courses) and 148 patients treated by primary surgery (PS). In the NAC group, the overall response rate from MVAC was 88.1 per cent with 31.0 per cent having complete clinical response and 7.1 per cent with complete pathological response. Pelvic lymph node metastasis was not significantly different between the NAC group (16.7%) and the PS group (18.2%). At a median follow-up of 64.5 months, 19.0 per cent in the NAC group and 18.2 per cent in the PS group had tumor recurrence. The 5-year progression free and overall survival was 80.8 per cent and 92.0 per cent respectively for the NAC group which was not significantly different from 80.2 per cent and 92.9 per cent respectively in the PS group. In conclusion, although NAC can decrease the tumor size and produce a high response rate, it does not improve survival in bulky stage IB and IIA cervical cancer patients. PMID- 11853299 TI - The outcome of sperm retrieval and intracytoplasmic sperm injection for obstructive azoospermia. AB - OBJECTIVES: To study the outcome of sperm retrieval and intracytoplasmic sperm injection (ICSI) from obstructive azoospermic men. METHOD: Overall, 50 sperm retrieval procedures were performed in 45 obstructive azoospermic men, followed by 57 ICSI procedures with fresh epididymal spermatozoa (n=40), fresh testicular spermatozoa (n=4) or frozen-thawed epididymal spermatozoa (n=13). RESULTS: Sperm retrieval was accomplished via percutaneous epididymal sperm aspiration (PESA) in 42 cases, testicular sperm aspiration (TESA) in 1 case and testicular sperm extraction (TESE) in 2 cases. TESA and TESE were only applied when PESA failed to produce enough spermatozoa for simultaneous ICSI. PESA was successful in 92 per cent of cases. Fertilization rate after ICSI was 79.6 per cent of the metaphase II oocytes. Seventy one embryo transfers were performed using both fresh and frozen thawed embryos resulting in clinical pregnancy in 39.4 per cent. Ongoing pregnancy was achieved in 35.2 per cent. CONCLUSION: ICSI has been shown to give a high fertilization and pregnancy rate with epididymal and testicular spermatozoa retrieved from obstructive azoospermic men. PESA is a noninvasive and simple technique for retrieving spermatozoa from obstructive azoospermic men. Therefore, it is suitable as the primary sperm recovery technique in patients with obstructive azoospermia. PMID- 11853300 TI - Risk factors of hip fracture among Thai female patients. AB - A case-control study on hip fracture among female patients, aged 50 years and over, were conducted from 1995 to 1996. A consecutive series of newly admitted female patients with hip fracture (n = 60) and 60 age-matched controls were recruited. Four independent risk factors of hip fracture were identified, including "type of principle occupation as sedentary or housework", "low number of pregnancies", "low serum albumin" and "low bone mass density at inter trochanter of the femur". When all univariate factors except bone density variables were entered into a multiple logistic regression analysis, four independent factors were identified including "no intake of milk regularly", "low number of pregnancies", "thin body appearance", and "low serum calcium". The study confirms the role of calcium intake as a preventive measure of hip fracture. Women with low weight-bearing work and housewives should have regular weight-bearing exercise. PMID- 11853301 TI - Effect of lower uterine segment sweeping on progress of labor in nullipara. AB - Previous studies have shown that sweeping between the membranes and lower uterine segment was an effective procedure for reducing prolonged pregnancy. However, there has been no study to show the effect of lower uterine segment sweeping to the progress of the active phase of labor. This study was to determine the effect of lower uterine segment sweeping on the progress of the active phase of labor in nullipara. A total of four hundred nulliparous term pregnant women with spontaneous labor were randomized to one of two groups; the control group had routine vaginal examination while the study group had lower uterine segment sweeping at the time of each examination. Oxytocin infusion was given if there was dysfunctional labor. Progression of labor and the need for oxytocin augmentation were reviewed. Data were analyzed by chi square and Student t-test. Oxytocin was used in 67 per cent of the sweeping group and 62 per cent of the control group (p=0.3). The duration of the first stage (0.46), the second stage (0.38), and the third stage (0.28) of labor were not significantly different between the two groups. In conclusion, lower uterine segment sweeping did not reduce the need for oxytocin augmentation or lessen the duration of labor in nullipara. PMID- 11853302 TI - Self-evaluation of obstetricians by delivery data to reduce cesarean section rate in Chai Nat Hospital. AB - INTRODUCTION: The cesarean section rate has continuously increased in recent years in both public and private sector services. Various strategies have been proposed to reduce the cesarean section rate, however, none of them has proven to be effective. Cesarean section rate at Chai Nat Hospital climbed from 9.69 per cent in 1987 to 31.13 per cent in 1997. The authors have attempted to reduce the unnecessary rate by creating a strategy of self-evaluation of obstetricians by delivery data of each obstetrician. The objective of this study was to evaluate the effectiveness of the strategy by comparing the cesarean section rate before and after using the strategy with the assumption that the cesarean section rate would probably be decreased with the strategy. PATIENTS AND METHOD: The study was a clinical experimental research conducted from January 1995 to December 1999, a sixty-month period. The target populations included obstetricians and pregnant women delivering at Chai Nat Hospital. The cesarean section rate during the first thirty months was compared with that of the latter thirty months, the period in which the strategy was used. Furthermore, the cesarean section rate was also compared with that in Phra Phutthabat Hospital. RESULTS: The cesarean section rate in the first half of the study (30 months) at Chai Nat Hospital, the period without the strategy, among 4,843 deliveries was 28.78 per cent compared with 25.40 per cent among 5,044 cases in the second half of the study, the period using the strategy. The rate was significantly decreased (Z=3.79, P<0.001). At Phra Phutthabat Hospital where the strategy was not used, the cesarean section rate during the first 30-month period was 29.36 per cent among 5,868 deliveries and 28.12 per cent among 6,020 deliveries during the second half of the study. The rates were not significantly different (Z=1.49, P>0.05). The comparison of cesarean section rates during the first 30-months between Chai Nat and Phra Phutthabat Hospital during the first half showed no significant difference (Z=0.66, P>0.05), whereas that during the second half of the study between both hospitals was significantly different (Z=3.22, P<0.01). CONCLUSIONS: The strategy of self-evaluation of obstetricians by delivery data of each obstetrician could significantly reduce the cesarean section rate (P<0.001). If the strategy is implemented in larger areas of Thailand, a large number of unnecessary cesarean sections as well as maternal morbidity could be avoided, resulting in saving a lot of national expense and also improving the standard of care in obstetrics. PMID- 11853304 TI - Isomeric separation of methamphetamine by HPLC chiral column. AB - Methamphetamine and its active metabolite, amphetamine, are optically active compounds which, based upon synthetic routes, can be found in two forms; pure d form and racemic mixture. Analysis of their isomers can help to identify which precursor is currently spreading widely in a given region. Since there are many drugs that can be metabolized to amphetamine/methamphetamine, isomeric separation can be a useful tool for evaluation of these drugs, as well. Indirect method by using N-trifluoroacetyl-1-prolyl chloride (1-TPC) was found to have limited accuracy due to the contribution effect. In this presentation a direct method using HPLC Chirex chiral column 3022 was studied. Although the method gave no base-line separation of two different isomer peaks, it gave good sensitivity, reliability, and linearity. No contribution effect was found in the method presented. It also gave excellent correlation with the 1-TPC method. PMID- 11853303 TI - Invasive group A Streptococcal infections at Chulalongkorn University Hospital. AB - To determine whether the incidence and pattern of group A Streptococcal (GAS) infections in Thailand have paralleled those in the United States and Europe, we conducted a retrospective study of invasive GAS infections at Chulalongkorn University Hospital from 1995 to 1999. A total of 42 cases were identified. There were 18 males and 24 females (median age of 59 and 46 years, respectively). Most patients were in two age groups: 20-39 (33%) and 60-79 (38%). Underlying conditions were present in 34 patients (81%), including mostly chronic system diseases (50%), alcohol abuse (19%), diabetes mellitus (14%), connective tissue diseases (12%), immunosuppressive illnesses (12%), and human immunodeficiency virus infection (10%). The most common clinical presentations were skin and soft tissue infections (31%), primary bacteremia (29%), and arthritis (14%). Of these, 24 (57%) presented with toxic shock syndrome (TSS). Overall mortality rate was 33 per cent. All GAS but one isolate were susceptible to penicillin. PMID- 11853305 TI - Teaching instrument: a laparoscopic training model. AB - A laparoscopic training model with female surface anatomy has been developed. This training model is made of a plastic boutique-showing model that is equivalent to normal-size female anatomy from the neck to the upper thigh. Four holes were made on the model's abdominal wall as puncture-sites to enter the cavity, the first is 80-mm diameter at the umbilical area, and the other three 38 mm diameter holes are located on both sides of the lower abdomen and suprapubic area. The umbilical hole can be covered with a simulated abdominal wall made from 6.5-mm insulation sheet, fixed to the model using a rubber band. The other three puncture-sites were plugged with a flexible rubber diaphragm as working ports. When used as video-laparoscopy, the auto-focus camcorder is used as a telescope and is connected to a regular television set as a monitor. This model can be used for training of abdominal entry by Veress needle or trocar, laparoscopic tubal ligation (LTL), and video-eye-hand co-ordination. This model has been a training medium in our Department for 1 year and was included in the OSCE for the Board Examination of the Royal Thai College of Obstetricians and Gynecologists in the year 2000 to assess the process of Veress needle insertion. PMID- 11853306 TI - Ultrastructural study of glomus tumor. AB - The fine structure of five cases of glomus tumor was described. All of the cases showed a solid type, and the glomus cells were shown to be typical or modified smooth muscle cells. The authors' conclusion is that the glomus cells were not derived from pericytes, but from smooth muscle cells of vascular part of the glomus, and are thought to be a specialized smooth muscle cell tumor. PMID- 11853307 TI - Quetiapine for tic disorder: a case report. AB - Tic disorders happen in nearly 20 per cent of children. There is no "best drug" to treat this illness. Potent antipsychotics e.g. haloperidol and pimozide, are the most effective drugs but their limitations are their extrapyramidal side effects (EPS). Risperidone has been proved on efficacy for tic disorders but EPS still remain, even though it was claimed to be less. Thus, quetiapine, a newer atypical neuroleptic with the same action as risperidone and produces fewer EPS, was included in this study. OBJECTIVE: To study the efficacy and side effects of quetiapine in tic disorders. METHOD: A case report of a 19-year-old female patient with tic disorder who had taken haloperidol 2 mg/d with benzhexol HCl 2-4 mg/d, then switched to risperidone 1.5 mg/d with benzhexol HCl 4 mg/d because of acute dystonia and oculogyric. She was then prescribed quetiapine, 50 mg/d as a starting dose without benzhexol HCI, because of the remaining symptoms and EPS. The severity of the symptoms was assessed monthly using the Behavior Rating Scale. The dose was increased by 50 mg/d weekly for a better outcome. RESULTS: The tic was improved after the first week and disappeared for three weeks with 150 mg/d of quetiapine. However, the tic returned again, but less frequently (20%). Thus, the dose was stepped up to 200 mg/d. One week later, the patient reported that the tic has disappeared. CONCLUSION: Quetiapine showed the efficacy and fewest EPS in this patient. However, a further clinically controlled trial must be carried out before quetiapine can become the first-line treatment for tic disorders. PMID- 11853308 TI - Scalp replantation: a case report of long ischemic time. AB - A totally avulsed scalp was treated successfully by microvascular replantation in 1975 (1). Since then, scalp replantation has become the treatment of choice in scalp injuries and should always be considered even in cases of severe damage or prolonged ischemic time. Usually, every attempt is made to establish as many vascular anastomoses as possible. We report one case of total scalp avulsion with a 24-hour ischemic time, which was successfully replanted via one arterial and one venous anastomose. PMID- 11853309 TI - Presumably entomophthoramycosis in an HIV-infected patient: the first in Thailand. AB - The authors reported the case of a symptomatic HIV-infected woman with a slowly progressive infiltrative lesion which invaded in and around the nasal cavity over a 6-month period. Physical examination showed erythematous to violaceous plaques at the nasal and malar areas. Swelling of the inferior turbinate was noted in the right nare. Skin biopsy of the involved area revealed multiple nonseptate, broad, thin-walled hyphae within giant cells and granulomata. Entomophthoramycosis was diagnosed based on clinical features and histopathology. She was treated with intravenous amphotericin B for two weeks, followed by oral itraconazole 400 mg daily. At six months there was complete resolution of all lesions. PMID- 11853310 TI - Primary intraosseous glomus tumor: a case report. AB - We reported a rare case of a primary intraosseous glomus tumor in a 53-year-old woman who had a small lytic lesion in the distal phalanx of her left index finger. The radiologic appearance showed a well circumscribed osteolytic lesion without sclerotic rim. Histologic examination revealed solid nests or sheets of polygonal cells surrounding branching vasculatures consistent with a glomus tumor. Curettage resulted in complete removal of the tumor as well as pain alleviation. The patient was well without evidence of recurrent disease following an 18 month follow-up. Despite its rarity, intraosseous glomus tumor should be included in the differential diagnosis of bone lesions in which plain radiography showed a well circumscribed punch-out lesion without sclerotic rim especially those arising in the finger. PMID- 11853311 TI - Traumatic elbow subluxation in a child: case report. AB - An 11-year-old boy fell on his outstretched right arm, following which he had pain on the lateral aspect of his right elbow with limitation of range of motion. Roentgenograms showed right elbow subluxation. The elbow was reduced and immobilized in a long arm splint for 5 days. The result was excellent. Elbow subluxation in previous reports was the result of recurrent elbow dislocation with instability. There are no reports of elbow subluxation in patients who did not have underlying recurrent elbow dislocation or elbow instability. This condition is easy to diagnose and has an excellent outcome. PMID- 11853312 TI - New-world and transcultural impact on Thai medical practices and professional behaviour. AB - Thailand, which is barely recovering from the 1997 economic collapse, has passed a new constitution, has a new government and all of this will have a profound effect on the way doctors practice and interact with their patients. Some of the factors that have been responsible for this are 1) Passage of a patient's rights bill 2) Legal definition of what is a proper consent form 3) The public's demand for openness, better communication and transparency in medicine These concepts, which now have a legal as well as moral basis, will impact directly on doctors and nurses as their practices will be subject to increasingly intense scrutiny and criticism. Doctors and nurses in past generations were among the most respected members of Thai society. They were trusted almost without exception and their judgment was not questioned. Only rare cases of malfeasance and malpractice came to public notice though, no doubt, they existed but to a much lesser degree. A physicians' life was much simpler for our teachers, fathers and grandfathers. Life was inexpensive, families were less demanding and the doctor did not have to surround himself with expensive foreign-made apparatus which can bankrupt the doctor as well as the patients who have to pay the bills. Medical decisions were made mostly on the basis of observation, past experience at the bedside and not in the laboratory or imaging rooms. Honest errors in diagnosis were common, treatment was limited and simple and this was all tolerated or expected by the public. Few if any disappointments on the part of patients ended up before a judge. PMID- 11853313 TI - Comparison of sequences of cDNA clones obtained from oligo-capping cDNA libraries with those from unigene. AB - We compared in detail the characteristics of the sequences of the cDNA clones obtained by the oligo-capping method (oligo-capping clones) with that of the sequences in the UniGene database. To compare the completeness of the sequences, three new variables, "fullness-proportion of clones" (the ratio of complete clones to total clones in a library), "fullness-proportion of genes" (the ratio of complete genes to total genes in a library), and "fullness-proportion of database" (the ratio of complete genes to total genes in a database sampled from a library), were defined. The fullness-proportion of clones of oligo-capping clones was 57.3%, 2.2 times larger than that of UniGene (25.9%). The fullness proportion of genes of oligo-capping clones was 41.8%, 2.4 times larger than that of UniGene (17.8%). When gene length was restricted to > or = 1.5 kb, the fullness-proportion of genes of oligo-capping clones was four times larger than that of UniGene. The fullness-proportion of database of oligo-capping clones was approximately the same as that of UniGene. By simulating the clone redundancy, this coincidence was found to be due to the large redundancy of the UniGene database. Consequently, the cDNA sequence database of oligo-capping clones enabled high throughput selection of full-length cDNA clones. PMID- 11853314 TI - Cloning and sequencing of the beta-glucosidase gene from Acetobacter xylinum ATCC 23769. AB - The beta-glucosidase gene (bglxA) was cloned from the genomic DNA of Acetobacter xylinum ATCC 23769 and its nucleotide sequence (2200 bp) was determined. This bglxA gene was present downstream of the cellulose synthase operon and coded for a polypeptide of molecular mass 79 kDa. The overexpression of the beta glucosidase in A. xylinum caused a tenfold increase in activity compared to the wild-type strain. In addition, the action pattern of the enzyme was identified as G3ase activity. The deduced amino acid sequence of the bglxA gene showed 72.3%, 49.6%, and 45.1% identity with the beta-glucosidases from A. xylinum subsp. sucrofermentans, Cellvibrio gilvus, and Mycobacterium tuberculosis, respectively. Based on amino acid sequence similarities, the beta-glucosidase (BglxA) was assigned to family 3 of the glycosyl hydrolases. PMID- 11853315 TI - The size and sequence organization of the centromeric region of Arabidopsis thaliana chromosome 4. AB - We have determined the genome structure of the centromeric region of Arabidopsis thaliana chromosome 4 by sequence analysis of BAC clones obtained by genome walking, followed by construction of a physical map using DNA of a hypomethylated strain. The total size of the centromeric region, corresponding to the recombinant inbred (RI) markers between mi87 and mi167, was approximately 5.3 megabases (Mb). This value is over 3 Mb longer than that previously estimated by the Arabidopsis Genome Initiative (Nature, 408, 796-815, 2000). Although we could not cover the entire centromeric region by BAC clones because of the presence of highly repetitive sequences in the middle (2.7 Mb), the cloned regions spanning approximately 1 Mb at both sides of the gap were newly sequenced. These results together with the reported sequences in the adjacent regions suggest that the centromeric region is principally composed of a central domain of 2.7 Mb, consisting of mainly 180-bp repeats and Athila elements, and upper and lower flanking regions of 1.55 Mb and 1 Mb, respectively. The flanking regions were predominantly composed of various types of transposable elements, except for the upper end moiety in which a large 5S rDNA array (0.65 Mb) and central domain-like sequence are present. Such an organization is essentially identical to the centromeric region of chromosome 5 reported previously. PMID- 11853316 TI - A new class of LINEs (ATLN-L) from Arabidopsis thaliana with extraordinary structural features. AB - The Arabidopsis thaliana genome has about 250 copies of LINEs (here called ATLNs). Of these, some, called ATLN-Ls, have an extra sequence of about 2 kb in the region downstream of two consecutive open reading frames, orf1 and orf2. Interestingly, the extra sequences in these ATLN-L members have another open reading frame, designated as orf3. Each member is flanked by direct repeats of a target site sequence, showing that ATLN-L members with the three open reading frames have retrotransposed as a unit. The ATLN-L members are also distinct from other ATLN members: orf1 terminates with TAA (or TAG) and is located in the same frame as orf2, and the ATG initiation codon of orf2 is not present in the proximal region. A sequence that may form a pseudoknot structure in ATLN-L mRNA was present in the proximal region of orf2, therefore the TAA (or TAG) termination codon of orf1 is assumed to be suppressed to produce an Orf1-Orf2 transframe protein during the translation of the ATLN-L mRNA. The region between orf2 and orf3 is several hundred bp long, suggesting that orf3 expression is independent of orfl-orf2. The amino acid sequences of the proteins Orf1 and Orf3 are highly homologous in their N-terminal half regions that have a retroviral zinc-finger motif for RNA binding. Orf3, however, has a leucine-zipper motif in addition to the zinc-finger motif. The C-terminal regions of the Orf1 and Orf3 proteins have poor homology, but seem to have nuclear localization signals, suggesting that these proteins are involved in the transfer of ATLN-L mRNA to nuclei. A phylogenetic tree shows that Orf3 proteins form a branch distinct from the branches of the Orf1 proteins encoded by ATLN-L members. This indicates that an ancestor element of ATLN-Ls has incorporated the orf1 frame carried by another ATLN member into its distal region to orf1-orf2 during evolution. PMID- 11853317 TI - Construction of a genetic linkage map of the model legume Lotus japonicus using an intraspecific F2 population. AB - Among leguminous plants, the model legume Lotus japonicus (Regel) Larsen has many biological and genetic advantages. We have developed a genetic linkage map of L. japonicus based on amplified fragment length polymorphism (AFLP), simple sequence repeat polymorphism (SSRP) and derived cleaved amplified polymorphic sequence (dCAPS). The F2 mapping population used was derived from a cross between two L. japonicus accessions Gifu B-129 and Miyakojima MG-20. These parental accessions showed remarkable cytological differences, particularly with respect to size and morphology of chromosomes 1 and 2. Using fluorescence in situ hybridization (FISH) with BAC clones from Gifu B-129 and TAC (Transformation-competent Artificial Chromosome) clones from Miyakojima MG-20, a reciprocal translocation was found to be responsible for the cytological differences between chromosomes 1 and 2. The borders of the translocations were identified by FISH and by alignment toward the L. filicaulis x L. japonicus Gifu B-129 linkage map. The markers from the main translocated region were located on linkage groups 1 and 2 of the two accessions, Gifu B-129 and Miyakojima MG-20, respectively. The framework of the linkage map was constructed based on codominant markers, and then dominant markers were integrated separately in each linkage group of the parents. The resulting linkage groups correspond to the six pairs of chromosomes of L. japonicus and consist of 287 markers with 487.3 cM length in Gifu B-129 and 277 markers with 481.6 cM length in Miyakojima MG-20. The map and marker information is available through the World Wide Web at http://www.kazusa.or.jp/lotus/. PMID- 11853318 TI - Structural analysis of a Lotus japonicus genome. I. Sequence features and mapping of fifty-six TAC clones which cover the 5.4 mb regions of the genome. AB - A total of 56 TAC clones with an average insert size of 100 kb were isolated from a TAC library of the Lotus japonicus genome based on the expressed sequences tags (ESTs), cDNA and gene information, and their nucleotide sequences were determined according to the shot-gun based strategy. The total length of the sequenced regions is 5,473,195 bp. By comparison with the sequences in protein and EST databases and analysis with computer programs for gene modeling, a total of 605 potential protein-encoding genes with known or predicted functions, 69 gene segments, and 172 pseudogenes were identified. The average density of the genes assigned so far is 1 gene/8120 bp. Introns were identified in approximately 78% of the potential genes. There was an average of 3.8 introns per gene and the average length of the introns was 375 bp. DNA markers were generated based on the nucleotide sequences obtained, and each clone was mapped onto the linkage map using the F2 mapping population derived from a cross of L. japonicus Gifu B-129 and Miyakojima MG-20. The sequence data, gene information and mapping information are available through the World Wide Web at http://www.kazusa.or.jp/lotus/. PMID- 11853319 TI - Prediction of the coding sequences of unidentified human genes. XXII. The complete sequences of 50 new cDNA clones which code for large proteins. AB - As an extension of human cDNA projects for accumulating sequence information on the coding sequences of unidentified genes, we herein present the entire sequences of 50 cDNA clones, named KIAA1939-KIAA1988. cDNA clones to be entirely sequenced were selected by two approaches based on their protein-coding potentialities prior to sequencing: 10 cDNA clones were chosen because their encoding proteins had a molecular mass larger than 50 kDa in an in vitro transcription/translation system; the remaining 40 cDNA clones were selected because their putative proteins-as determined by analysis of the genomic sequences flanked by both the terminal sequences of cDNAs using the GENSCAN gene prediction program-were larger than 400 amino acid residues. According to the sequence data, the average sizes of the inserts and corresponding open reading frames of cDNA clones analyzed here were 4.6 kb and 1.9 kb (643 amino acid residues), respectively. From the results of homology and motif searches against the public databases, the functional categories of the 31 predicted gene products could be assigned; 25 of these predicted gene products (81%) were classified into proteins relating to cell signaling/communication, nucleic acid management, and cell structure/motility. The expression profiles of the genes were also studied in 10 human tissues, 8 brain regions, spinal cord, fetal brain and fetal liver by reverse transcription-coupled polymerase chain reaction, the products of which were quantified by enzyme-linked immunosorbent assay. PMID- 11853320 TI - Vestibular evoked myogenic potentials: test-retest reliability. AB - Vestibular evoked myogenic potentials (VEMPs) are myogenic responses induced by stimulation of the saccular macula by intense sound stimuli. The responses are recordable from the sternocleidomastoid (SCM) muscles. We recorded VEMPs from normal subjects (up to three times in each subject) to identify: i) the best recording procedures, ii) the reliability, and iii) the normal limits for both individual point and test-retest evaluation. We adopted a recording setting in which the subjects were asked to simultaneously activate both SCM muscles by pushing their forehead against a load cell during a bilateral acoustic stimulation. This system enabled subjects to monitor their intensity of SCM activation and to keep intensity constant; us to record VEMPs from both sides simultaneously, and thus to minimize the duration of the recording session. For each subject we considered the mean and the difference (divided by the mean) of the values derived from the two SCM muscles of the latency of the P13 and N23 components and of the P13-N23 peak-to-peak amplitude. Reliability was evaluated by estimate of the intraclass correlation coefficient, and was good or excellent for all parameters, with the exception of the P13-N23 amplitude side-difference. To take advantage of all the data available, we computed the normal limits for both individual point and test-retest evaluation by means of the variability indices used for the evaluation of reliability. In this system, VEMP recording is simple, inexpensive and rapid. It is well tolerated by subjects, and easily implemented in laboratories equipped for evoked potential recording. PMID- 11853321 TI - Functional improvement in patients with severe spinal spasticity treated with chronic intrathecal baclofen infusion. AB - In this retrospective study we evaluated the efficacy and functional benefits of chronic intrathecal baclofen infusion in severe spinal spasticity. Twenty patients with a diagnosis of severe intractable spinal spasticity were evaluated prior to implantation of a programmable pump for chronic intrathecal baclofen therapy and at follow up, which ranged from 12 to 36 months (mean 22.4 months). The mean age of the patients was 39.1 years. The prevailing pathology was multiple sclerosis. All were unable to walk. Patient assessment was based on the Ashworth Scale, the Spasms Frequency Scale, self-reported pain and Functional Independence Measure (FIM) scores. The Wilcoxon test was used for statistical analysis. A statistically significant decrease in muscle tone, spasms and pain was observed in all the patients. The Ashworth score decreased from 4.4 to 1.8, the spasms frequency score from 2.5 to 0.5 and the self-reported pain score from 5.5 to 2.3. The FIM score also showed a statistically significant change (increasing from a mean of 33.8 to 58.7). Two patients in employment were able to return to work. No severe side effects were observed. Chronic intrathecal baclofen infusion was seen to produce a functional improvement in patients with severe spinal spasticity, particularly as regards bathing, comfortable wheelchair sitting and mobility. PMID- 11853322 TI - Short-term effects of "botulinum toxin a" as treatment for children with cerebral palsy: kinematic and kinetic aspects at the ankle joint. AB - Botulinum toxin A (BTA) therapy plays several roles in the management of paediatric cerebral palsy (CP). However, few studies contain objective documentation of gait changes. The main aim of this study was to provide objective information on the outcome of the treatment. Gait analysis data from 20 normal subjects and 23 CP children were collected before and after BTA injections into the gastrocnemius-soleus complex. The follow up was performed 1 month after the first injection. The kinematic and kinetic data revealed significant improvements in dynamic ankle dorsiflexion, both in stance and in the swing phase, an improvement of equinus foot upon initial contact and better support in stance. The results of this study are promising, but studies of other joints involved in gait, such as the knee, are also needed. PMID- 11853323 TI - Electroencephalographic findings in children with cerebral palsy: a study of 151 patients. AB - EEG abnormalities were studied in 151 patients (79 boys, 72 girls age range 0.4 13 years) with cerebral palsy (CP). They all had standardised EEG recordings, which were read by the same electroencephalographer. Eighty-one children had seizures and 70 were seizure-free. The EEG abnormalities in the seizure group included slow waves in 36 patients (generalised asynchronous in 33 and generalised synchronous in 3); amplitude abnormalities in 2 (focal in 1, generalised 1); epileptiform activity (including isolated sharp waves, isolated spikes, and spike-wave and polyspike-wave complexes) was seen in 66 (focal in 12; generalised in 48 and multifocal in 6). Hypsarrythmia was found in 4 and burst suppression in 1. Only 6 recordings were normal giving an overall percentage of abnormality of 92.6%. Of the CP patients without seizures, 28 (40%) showed generalised asynchronous slow waves; epileptiform activity was found in 27 (focal in 2, generalised in 23 and multifocal in 2); 3 subjects showed hypsarrythmia and 24 recordings were normal. The overall percentage of abnormality in this group was 76%. Cerebral palsy in children, regardless of its cause may be associated with generalised focal EEG abnormalities. This may reflect heterogeneity of the neural-generator in the underlying disease process. PMID- 11853324 TI - Neuropsychological changes after carotid endarterectomy. AB - Carotid endarterectomy (CEA), performed to prevent stroke, could lead to changes in cognitive functions. Sixty-four patients with severe carotid stenosis undergoing CEA treatment were evaluated by means of a detailed neuropsychological assessment before (baseline), from one to two weeks (1st follow up) after and 3 months (2nd follow up) after surgical operation. A significant post-CEA improvement was found in verbal memory and attention (p<0.05), while other cognitive functions showed no significant changes. PMID- 11853325 TI - Pelizaeus-Merzbacher disease: electrophysiological study of two sibs with the classic form and of their relatives. AB - We examined two sibs with the classic form of Pelizaeus-Merzbacher disease (PMD) and their relatives. Electromyographic-electroneurographic studies and magnetic stimulation of motor pathways were performed. In both patients we found an absence of compound motor action potential (cMAP) after stimulation of the motor cortex and a normal conduction time by stimulating the cervical roots. Despite reported sparing of the peripheral nervous system in PMD, our conduction study of the tibial nerve revealed a slightly decreased motor nerve conduction velocity in one patient. In both patients the EMG study showed neurogenic findings. The elder sister showed a prolonged central motor conduction time. This study demonstrates abnormalities of motor corticospinal pathways also in PMD relatives suggesting that magnetic stimulation could be useful in detecting "subclinical" abnormalities in this dysmyelinating condition. Furthermore, in accordance with previous studies, we suggest that a slight involvement of the peripheral nervous system could be observed in PMD. PMID- 11853326 TI - Evidence for an anatomical substrate of hyperexcitability in human temporal lobe epilepsy: glutamate receptor alterations and reorganized circuitry. PMID- 11853327 TI - Defining childhood obesity: fiddling whilst Rome burns? PMID- 11853329 TI - Sleep in school-age children. PMID- 11853328 TI - Is LBW a risk factor for linguistic development? PMID- 11853330 TI - Malabsorption of modified food starch (acetylated distarch phosphate) in normal infants and in 8-24-month-old toddlers with non-specific diarrhea, as influenced by sorbitol and fructose. AB - Acetylated distarch phosphate (ADiSP) is a modified starch used in some baby foods. The bioavailability of ADiSP and a native (unmodified) starch was evaluated in 20 normal infants and 21 toddlers aged 8-24 mo with chronic non specific diarrhea. Formulae contained 8% native or 8% modified waxy maize starch. No infant or toddler consuming Formula N (native starch) had elevated peak breath hydrogen levels (20 ppm or greater), stools clinically positive for reducing substances (0.75% or greater) or loose stools. Fourteen infants received formula M (modified starch): 2 had elevated breath hydrogen, 1 had positive stools and another had loose stools. Of the 21 toddlers fed formula M, 2 had elevated breath hydrogen, but none had positive stools or loose stools. Formula NS (native starch with 2% sorbitol) had little effect on breath hydrogen in the infants but significantly increased it in the toddlers. Formula NS produced loose stools in 2 toddlers but no clinically positive stools in any infant or toddler. Formula MS (modified starch with 2% sorbitol) elevated breath hydrogen in 3 infants and 8 toddlers, and produced positive stools in 2 infants and 2 toddlers, and loose stools in 4 infants and 7 toddlers. Formula MSF (modified starch with 2% sorbitol and 5% fructose) elevated breath hydrogen in 7 infants and 10 toddlers, positive stools in 7 infants and 6 toddlers, and loose stools or diarrhea in 7 infants and 11 toddlers. CONCLUSION: ADiSP modified starch can increase breath hydrogen and produce loose stools. Sorbitol and fructose aggravate the malabsorption, in some cases leading to frank diarrhea. PMID- 11853331 TI - Hyperimmune bovine colostrum in the treatment of shigellosis in children: a double-blind, randomized, controlled trial. AB - Immunological approaches have been considered as an alternative therapeutic option for the treatment of enteric infections over the past few years. Hyperimmune bovine colostrum (HBC) is a potentially innovative immunological option in the management of shigellosis together with traditional antibiotic therapy. Children aged 1-12 y with a history of bloody mucoid diarrhoea of less than 5 d duration were enrolled after their stool specimen was found to be positive for Shigella dysenteriae type I antigen by a rapid diagnostic fluorescent antibody staining test. They were randomized to receive either HBC containing very high titres of antibody against S. dysenteriae type I antigen or bovine colostrum (BC) without any antibody. The study group received 100 ml of HBC three times a day orally for 3 d and control group received BC. Children also received pivmecillinam in a dose of 50 mg kg(-1) d(-1) in four divided doses orally for 5 d. Admission characteristics of the 34 children in the HBC group and 35 in the BC group were comparable. No significant differences were observed in duration of diarrhoea, fever, anorexia, abdominal pain, tenesmus, stool frequency or visible blood in the stool between the groups. Two (6%) children in the study and five (14%) in the control group remained stool culture positive for S. dysenteriae type 1, even after 5 d of sensitive antimicrobial therapy. CONCLUSION: The results indicate that HBC as an adjuvant is unable to show any beneficial effect in reducing the stool frequency, duration or severity of childhood shigellosis due to S. dysenteriae type I infection. PMID- 11853332 TI - Sodium and water homeostasis in children with shigellosis. AB - Studies in Bangladesh have shown that the mortality in shigellosis is significantly higher in hyponatraemic (HN) than in normo- (NN) or hypernatraemic children. The aim of this study was to describe the effect of shigellosis on renal haemodynamics and sodium and water homeostasis before treatment was started. Twenty-one moderately ill children infected with Shigella dysenteriae type I were studied. Eight of them had a serum sodium concentration below 130 mmol/L. Renal function was determined by glomerular filtration rate measured by clearances of inulin and iohexol. Effective renal plasma flow was estimated by clearance of para-aminohippuric acid. Plasma renin, aldosterone and anti-diuretic hormone were also studied. The HN children had significantly higher haemoglobin and haematocrit levels than the NN group. There was an inverse correlation between serum sodium and haemoglobin, and a direct correlation between serum sodium and urinary sodium and urinary chloride. Direct correlations were found between serum aldosterone and haemoglobin, plasma renin and systolic blood pressure and an inverse correlation between serum aldosterone and serum sodium. Clearances of inulin and iohexol were normal. Detectable levels of ADH were found in both groups, despite low serum osmolalities. CONCLUSION: The HN state seems to be triggered by multiple factors. The normal glomerular filtration rate excludes a volume expansion secondary to reduced renal function. Inappropriate or a physiological increase of anti-diuretic hormone secretion may be of importance. The higher sodium losses in stools of the HN children might also be a factor contributing to the HN. PMID- 11853333 TI - Development of sucking behavior in infants with Down's syndrome. AB - The aim of this study was to gain a better understanding of the development of sucking behavior in infants with Down's syndrome. The sucking behavior of 14 infants with Down's syndrome was consecutively studied at 1, 4, 8 and 12 mo of age. They were free from complications that may cause sucking difficulty. The sucking pressure, expression pressure, frequency and duration were measured. In addition, an ultrasound study during sucking was performed in sagittal planes. Although levels of the sucking pressure and duration were in the normal range, significant development occurred with time. Ultrasonographic images showed deficiency in the smooth peristaltic tongue movement. CONCLUSION: The sucking deficiency in Down's syndrome may result from not only hypotonicity of the perioral muscles, lips and masticatory muscles, but also deficiency in the smooth tongue movement. This approach using the sucking pressure waveform and ultrasonography can help in the examination of the development of sucking behavior, intraoral movement and therapeutic effects. PMID- 11853334 TI - Thyroid dysfunction in children with Down's syndrome. AB - Thyroid function tests were carried out on 320 children with Down's syndrome aged between 5 d and 10 y. Thyroid function was normal in 230 patients (71.9%) and abnormal in 90 (28.1%). Six patients (1.8%) had primary congenital hypothyroidism, one patient had acquired hypothyroidism and two had transient hyperthyrotropinaemia of the newborn. Sixteen of the remaining 81 patients (25.3%) had compensated hypothyroidism with increased thyroid-stimulating hormone (TSH) levels (11-20 mU l(-1)). Their T4 levels were found to be either normal or close to the lower limit of normal. These cases were started on thyroxine therapy. Sixty-five of the 81 patients had a mild compensated hypothyroidism with mild TSH elevation (6-10 mU l(-1)). None of the patients had hyperthyroidism. The antithyroid antibodies were positive in the acquired hypothyroidism case. CONCLUSION: The prevalence of congenital hypothyroidism was 1.8% in children with Down's syndrome while 25.3% of them had compensated hypothyroidism. It is suggested that Down's syndrome patients with normal thyroid functions and those with compensated hypothyroidism should be followed annually and every 3 mo, respectively. Besides congenital hypothyroidism cases, those with TSH levels between 11 and 20 mU l(-1) may benefit from treatment with low-dose thyroxine. PMID- 11853335 TI - Real-time ultrasound-guided renal biopsy with a biopsy gun in children: safety and efficacy. AB - Real-time ultrasound-guided renal biopsy (RB) with a biopsy gun has become a standard procedure in the treatment of children. The purpose of the study was to establish the complication rate after real-time ultrasound-guided RB with a biopsy gun, the adequacy of renal tissue samples for pathohistological tests, the rate of concurrence between clinical and pathohistological diagnoses, and the benefits of the procedure. From January 1994 to October 1999, 88 renal biopsies were performed on 82 children, 81 of whom (35M, 46F, aged 3-20 y) were included in this retrospective study. The nephrotic syndrome (in infants, older children, those with evidence of nephritis or failing corticosteroid therapy) was the most frequent indication of RB. Other indications were non-nephrotic proteinuria, nephritic syndrome, glomerular haematuria, renal allograft dysfunction, unexplained acute or chronic renal failure, and kidney disease progression monitoring. No serious complications were noted. The adequacy rate of renal tissue samples ranged from 93.1 to 96.6%, depending on which definition of the adequacy of renal tissue samples was used. Clinical and pathohistological diagnoses matched in 81.4% of the cases. Data obtained by RB were very beneficial to patients in terms of establishing, confirming or altering the diagnosis and, consequently, the treatment. CONCLUSION: The results confirm that real-time ultrasound-guided RB with a biopsy gun is a safe procedure and provides information that is very beneficial to patients. PMID- 11853336 TI - Whole blood ionized magnesium in neonatal acidosis and preterm infants: a prospective consecutive study. AB - The mineral magnesium is a crucial enzymatic cofactor in the cellular bioenergetic process and alternations in magnesium metabolism may be associated with neurological impairment in newborn infants. Therefore, ionized magnesium (IMg) was measured in 14 newborn infants with acidosis [umbilical arterial cord pH 7.00 +/- 0.06, Apgar score 8.3 +/- 1.6 after 5 min, gestational age (GA) 276 +/- 16 d] and 15 premature infants (umbilical arterial cord pH 7.31 +/- 0.07, GA 236 +/- 12 d). Nineteen healthy mature infants served as controls. Arterial umbilical cord samples were taken immediately after delivery and capillary blood samples were taken 2, 6, 12 and 24 h after delivery by heel stick. IMg was measured by NOVA 8. The results showed an increased umbilical cord blood IMg in infants with acidosis compared with both premature and normal infants (0.58 +/- 0.08 mmol l(-1) vs 0.51 +/- 0.03 mmol l(-1) and 0.49 +/- 0.03 mmol l(-1); p < 0.0001). In infants with acidosis IMg declined significantly 2 h after delivery to 0.49 +/- 0.05 mmol l(-1) (p < 0.0001) and did not show any further significant changes during the first day of life. In premature infants and controls IMg levels were constant during the observation period. CONCLUSION: These findings suggest that elevated IMg is associated with neonatal acidosis. PMID- 11853337 TI - Computerized image analysis of bubbles in gastric aspirate for prediction of respiratory distress syndrome. AB - Gastric aspirates were obtained from 80 newborn babies (gestational age 24-34 wk). Fifteen of these babies developed respiratory distress syndrome (RDS). Bubbles were generated in the samples by vigorous suctioning and the number and size distribution of the bubbles were determined in random fields by computer aided image analysis. Sensitivity and specificity for prediction of RDS were 87% and 78% for average diameter of bubbles (cut-off points >45 microm) and similar for the proportion and total number of microbubbles (diameter <20 microm). CONCLUSION: Computerized image analysis of bubbles in gastric aspirate can be used for non-biased prediction of RDS in newborn babies. PMID- 11853338 TI - Urinary excretion of free cysteine in critically ill neonates. AB - In preliminary observations, significant amounts of free cysteine, a neurotoxic amino acid, were noted in the urine of asphyxiated or septic-shocked neonates. The present study was conducted to determine whether free urinary cysteine was elevated in these critically ill neonates compared with a control group, and to assess the clinical significance of this generation. Free cysteine was measured in the urine of newborn infants with perinatal asphyxia (n = 16) or neonatal sepsis (n = 14) and the urine of a control group (n = 10) by ion-exchange chromatography. Relationships between cysteine levels and the clinical severity, sulfite supply and neurological outcome of the patients were then studied. Urinary cysteine was 27.6 (15-49) mmol mol(-1) creatinine for the patients but was not detectable in the control group. Cysteine levels were correlated with the severity of neonatal septic shock but not with the grade of perinatal asphyxia and did not have a specific influence on the neurological outcome of these patients. The correlation between cysteine level and the severity of neonatal septic shock was indirect and probably linked to higher sulfite administration in this population. CONCLUSION: The mean daily supply of sulfites is high in critically ill neonates, mainly originating from dopamine and generating significant amounts of cysteine. Although a worsening effect attributable to cysteine on the neurological outcome of the patients could not be demonstrated, the appropriateness of cryptic administration of sulfites by way of drug excipients is called into question. PMID- 11853339 TI - Hearing impairment in preterm very low birthweight babies detected at term by brainstem auditory evoked responses. AB - Seventy preterm babies who were born with a birthweight <1500 g were studied with brainstem auditory evoked responses (BAER) at 37-42 wk of postconceptional age. The data were compared with those of normal term neonates to determine the prevalence of hearing impairment in preterm very low birthweight (VLBW) babies when they reached term. The BAER was recorded with click stimuli at 21 s(-1). Wave I and V latencies increased significantly (ANOVA p < 0.01 and 0.001). I-V and III-V intervals also increased significantly (p < 0.05 and 0.001). Wave V amplitude and V/I amplitude ratio did not differ significantly from those in the normal term controls. Ten of the 70 VLBW babies had a significant elevation in BAER threshold (>30 dB normal hearing level). Eleven had an increase in I-V interval (>2.5 SD above the mean in the normal controls) and one had a decrease in V/I amplitude ratio (<0.45). These results suggest that 14% (10/70) of the VLBW babies had a peripheral hearing impairment and 17% (12/70) a central impairment. Three babies had both an increase in I-V interval and an elevation in BAER threshold, suggesting that 4% (3/70) had both peripheral and central impairments. Thus, the total prevalence of hearing impairment was 27% (19/70). CONCLUSION: About one in four preterm VLBW babies has peripheral and/or central hearing impairment at term. VLBW and its associated unfavourable perinatal factors predispose the babies to hearing impairment. PMID- 11853340 TI - Comparison of brainstem auditory evoked responses recorded at different presentation rates of clicks in term neonates after asphyxia. AB - This study examined whether high presentation rates of clicks while recording brainstem auditory evoked responses (BAER) can improve the detection of central auditory impairment in asphyxiated neonates using the BAER. The BAER was analysed at different presentation rates of clicks within the first week after birth in 38 term neonates who suffered perinatal asphyxia. At the routinely used 21 s(-1) clicks all BAER wave latencies increased significantly (ANOVA, p < 0.05-0.01). After excluding five neonates who had a significantly elevated BAER threshold, only wave V latency increased slightly (p < 0.05). The interpeak intervals of I-V and III-V also increased slightly (both p < 0.05). Similar results were found at 51 s(-1) clicks. As the clicks were increased to 91 s(-1), the III-V interval increased more significantly (p < 0.01) and the III-V/I-III interval ratio also increased significantly (p < 0.01). In particular, wave V amplitude reduced more significantly than that in normal term controls (p < 0.01). Compared with values in the controls, wave V amplitude reduced by 4.5%, 12.2% and 24.7% at 21, 51 and 91 s(-1) clicks, respectively. CONCLUSION: Although a moderate increase in the rate (e.g. 51 s(-1)) while recording the BAER did not improve the detection of hypoxic-ischaemic auditory impairment, a significant increase (e.g. 91 s(-1)) did, which mainly indicates an abnormal reduction in wave V amplitude. PMID- 11853341 TI - Vaccination rate and age of premature infants weighing <1500 g: a pilot study in north-western Switzerland. AB - In Switzerland, there are no special vaccination recommendations for premature and low-birthweight infants with respect to a particular target vaccination age. Incomplete and delayed vaccination bears the inherent risk of preventable infections. Therefore, the vaccination rate and age of 60 premature infants in north-western Switzerland born in 1994/95 were investigated in a retrospective case-control study. For this group of patients these are the first data ever available for this region. At the age of 4-5 y, the vaccination rates for polio and diphtheria, tetanus, pertussis (DTP acellular) as well as Haemophilus influenzae b (Hib) were similar in both preterm and full-term infants. In both groups, the fourth dose of vaccine against DTP, Hib and polio was far less frequently administered than the first three. The vaccination age in preterm infants for most vaccinations was significantly higher than in age-matched full term controls. This was particularly obvious for the first dose of vaccine against polio and DTP. In preterm infants, the median age (5th; 95th percentile) at the date of the first polio vaccination was 131 (89; 270) d and 82 (60; 182) in full-term controls (p < 0.00001). The age difference was even larger for the first DTP vaccination (62 d, p < 0.00001). The main reasons for delayed vaccination may include insufficient information given to parents as well as prolonged hospitalization. CONCLUSION: Vaccination of preterm infants should be discussed in every discharge communication, with emphasis on vaccine administration at the appropriate chronological age. PMID- 11853342 TI - Body mass index reference values (mean and SD) for Swedish children. AB - Body mass index (BMI) is an important indicator of nutritional status. Many studies have been done to present BMI reference values in centile values rather than mean and SD values since its statistical distribution is positively skewed. Both height and weight growth charts are usually available in terms of mean and 1, 2 and 3 SD around the means; it would be of clinical value to produce BMI reference charts in a similar way. The aim of this work was to derive the mean and +/- 1, 2 and 3 SD BMI reference ranges as a supplement to the BMI centile reference values published previously for the same group of Swedish children. The method was based on an age-dependent Box transformation, and the beta-value was given as a third-degree polynomial function over the paediatric age. The BMI reference values can be given from mathematical functions in addition to values for specific ages. CONCLUSION: The BMI reference values and charts derived as described effectively reflect the nature of the variant age-dependent positive skewed statistical distribution of BMI values in the population, and can serve as a valid supplementary tool in the evaluation of growth and nutrition during paediatric years. PMID- 11853343 TI - Prevalence and awareness of excess weight in 13 and 14 year olds in Northern Ireland using recent international guidelines. AB - Concern has been expressed in recent years over the rising trends in overweight and obesity, particularly in children in developed countries. No recent data were available for Northern Ireland, therefore measurements of height and weight were included in a study in Northern Ireland originally designed as part of an international study to estimate the prevalence of symptoms of asthma and atopy in childhood. Twenty-six schools across the province were sampled. 2484 children aged mainly 13 and 14 y completed questionnaires, and 2307 were weighed and measured. Using recently proposed international guidelines on defining overweight and obesity in children the prevalence of overweight in boys was 16%, with 4% being obese. In girls 16% were also overweight and 2% were obese. This concurs with figures for English and Scottish schoolchildren up to 12 y of age from 1994. Less than one-sixth of obese boys but more than one-third of obese girls were on weight-reducing diets (the majority self-prescribed), reflecting differing attitudes towards excess weight. CONCLUSION: In line with figures in younger children from England and Scotland this study shows that 16% of boys and girls from Northern Ireland are classed as overweight using recent international guidelines. Only 15% of obese boys compared with 38% of obese girls stated that they were on weight-reducing diets. PMID- 11853344 TI - Linguistic and motor abilities of low-birthweight children as assessed by parents and teachers at 8 years of age. AB - Linguistic and motor abilities among low-birthweight 8-y-old children in the northern Finland Birth Cohort for 1985-1986 (n = 9322) were studied using parental and teacher evaluations. The parents of 8370 (90%) and teachers of 8525 (92%) children returned a mailed questionnaire concerning the children's speech, language, learning and motor abilities. Low-birthweight (LBW, < 2500 g) children (n = 279) appeared to have experienced more difficulties than normal-birthweight (NBW, > or = 2500 g) children (n = 8091). The parents evaluated the LBW boys to be the poorest in linguistic and motor skills compared with the other boys or any of the groups of girls. They are therefore presumably at risk of having problems at school, which was confirmed by the teachers' reports. There was also a clear relationship between speech/linguistic and motor disabilities. CONCLUSION: Multivariate logistic regression analyses showed that the lower birthweight and some sociodemographic factors, for example the mother's age being between 20 and 24 y, having more than four children in the family, a reconstructed family, as well as hearing impairment and male gender were the most important determinants of poor speech and language abilities at 8 y of age, with and without adjustment for neonatal risk factors (asphyxia, convulsions, respiratory distress syndrome, bronchopulmonary dysplasia, patent ductus arteriosus). Smallness for gestational age was also a risk factor for poor speech and language skills. Preterm birth was associated with poor skills only after adjustment for the neonatal risk factor. The parental and teacher evaluations were concordant concerning the LBW children's outcome. PMID- 11853345 TI - Sleep habits and sleep problems among a community sample of schoolchildren. AB - Sleep habits, sleep problems and subjective depth of sleep among 1413 schoolchildren aged 6.2-10.9 y were examined via a questionnaire, answered by the child and parent together. Total sleep time was approximately 10.5 h, with no difference between the sexes. Of 887 children who reported that they were awoken at night, parents considered that 75% were superficial sleepers and 25% were deep sleepers. The prevalence of frequent insomnia, sleepwalking and daytime sleepiness was 13, 7 and 4%, respectively. Logistic regression analyses indicated that onset insomnia was associated with fear of sleeping alone, bone pains, hypnagogic myoclonias, rhythmic movement disorder, enuresis, nocturia, confusion when awoken at night, nightmares, bodily movements during sleep, interrupted sleep, daytime sleepiness and daytime headache or stomach ache. Somnambulism was associated with rhythmic movement disorder, somniloquy, spontaneous confused arousals, nocturia and confusion when awoken at night. Increased risk of daytime sleepiness was found among children with fear of sleeping alone, onset insomnia, rhythmic movement disorder, spontaneous confused arousals, snoring, confusion when awoken, nightmares, bodily movements during sleep and headache or stomach ache. CONCLUSION: The results support the notion that onset insomnia is a problem with a predominantly psychological and behavioural background, while sleepwalking is a disorder of arousal without major psychological implications. The mechanisms behind daytime sleepiness seem to be multifactorial. PMID- 11853346 TI - Short-term prospective study of sleep disturbances in 5-8-year-old children. AB - The course and psychological correlates of disturbed sleep during middle childhood were investigated using parents' responses to questionnaires. Five specified sleep disturbances were explored in 614 5-8-y-old children, at baseline and at follow-up, 14 +/- 3 mo later. Difficulties falling asleep, in 6.2% of the children at baseline, persisted in 47.4% of the cases at follow-up; night-waking, in 18.6% at baseline, persisted in 45.5%; snoring, in 9.4% at baseline, persisted in 60%; nightmares, in 4.4% at baseline, persisted in 29.6%, and bedwetting, in 6% at baseline, persisted in 46%. Moreover, children with difficulties falling asleep at baseline frequently displayed bedtime resistance and shorter sleep at follow-up, while night-waking at baseline was related to co-sleeping at follow up. Snoring at baseline was associated with a trend towards restless sleep at follow-up. Nightmares at baseline were related to several other sleep complaints as well as to behavioural difficulties during daytime at follow-up. Bedwetting at baseline was associated with tendencies towards hyperactivity at follow-up. Separate analyses of cases of persisting sleep disturbances showed that persisting difficulties falling asleep were associated with a need to remedy sleep problems, while persisting nightmares were strongly related to reports suggesting behavioural and emotional problems. CONCLUSION: During middle childhood, difficulties falling asleep, night-waking, snoring, nightmares or bedwetting commonly persist over the course of a year. Persistent nightmares frequently indicate significant psychological problems in affected children. PMID- 11853347 TI - Comparison of parenting stress between Malaysian mothers of four-year-old very low birthweight and normal birthweight children. AB - A study was carried out to compare parenting stress between 116 mothers of very low birthweight (VLBW) children and 96 mothers of normal birthweight (NBW) children at 4 y of age, using the Parenting Stress Index (PSI). Multiple regression analysis was used to determine factors associated with child-domain stress (CDS) and parent-domain stress (PDS). There was a significantly higher proportion (39.7%) of mothers of VLBW children with high CDS scores >90th percentile than mothers of NBW children (20.8%). No significant differences were observed for PDS scores. Lower intelligence quotient (IQ) scores and adverse child behaviour, as evidenced by higher Child Behavior Checklist (CBCL) scores, were significantly associated with higher CDS scores (p < 0.001). Factors associated with higher PDS scores were higher CBCL scores (p < 0.001), mothers who were the primary caregivers (p < 0.001), male sex (p = 0.018) and lower level of maternal education (p = 0.048). These factors remained statistically significant even when physically and cognitively impaired children were excluded from the analysis. CONCLUSION: Specific child characteristics and the social environment appear to have a greater impact on parenting stress than the biological risk of VLBW birth per se. PMID- 11853348 TI - The Chinese version of the Parenting Stress Index: a psychometric study. AB - This study examined the psychometric properties of a Chinese version of the Parenting Stress Index/Short Form (PSI/SF). A 15-item simplified PSI/SF (S PSI/SF) was subsequently developed which maintained a level of reliability and validity similar to the full version. The Chinese PSI/SF was tested on 149 parents (100 mothers, 49 fathers) of pediatric cancer patients in Taiwan. Psychometric testing was conducted using item analysis, Cronbach's alpha and confirmatory factor analysis. The S-PSI/SF was constructed based on the item analysis of the PSI/SF. Both the PSI/SF and S-PSI/SF produced good reliability coefficients. Confirmatory factor analyses indicated that both PSI/SF and S PSI/SF met all criteria for goodness of fit. Compared with the PSI/SF, the S PSI/SF demonstrated better internal consistency and overall fit at the one subscale level, and satisfactory overall fit at two- and three-subscale levels. Despite the limited number of items included, the S-PSI/SF had a very good factor structure. No gender difference in parenting distress index was observed between mothers and fathers of pediatric cancer patients. CONCLUSION: The 15-item S PSI/SF is a brief, easily administered instrument that has evidence of reliability and validity in Taiwanese parents of children with cancer. It could serve as a valuable assessment tool in clinical practice to identify parenting stress with a need for intervention. PMID- 11853349 TI - A new congenital myopathy in a Norwegian family. AB - A 6-y-old boy presented with a mild, and apparently non-progressive, congenital myopathy, primarily affecting explosive movements such as running and jumping. Five other cases, spanning four generations, were identified in his family. A dominant inheritance pattern was suggested. Quadriceps muscle histology showed a selective type II fibre atrophy, which is otherwise considered a non-specific change associated with a number of conditions. CONCLUSION: A Norwegian boy with an inherited muscle weakness is presented. Based on clinical and laboratory investigations, and in light of the inheritance pattern, a previously undescribed congenital myopathy is suggested. PMID- 11853350 TI - Pneumocystis carinii pneumonitis in haemophagocytic lymphohistiocytosis. AB - We report on a 10-y-old boy who developed Pneumocystis carinii pneumonitis (PCP) as the dominant symptom at the onset of haemophagocytic lymphohistiocytosis (HLH). PCP is a common infection in patients with combined primary immunodeficiencies or acquired immunodeficiency syndrome but it has rarely been observed at the onset of HLH. Typically, HLH presents as a febrile syndrome associated with cytopenia and hepatosplenomegaly. Repeated bone marrow aspirates, spleen or lymph node biopsies are sometimes required to reveal haemophagocytosis. Because of the significant immunosuppression during treatment of HLH, prophylaxis of PCP with co-trimoxazole is recommended. However, de-arranged immune response in HLH renders the patients susceptible to opportunistic infections, even before the introduction of immunosuppressants. CONCLUSION: We suggest that in patients with unclear respiratory symptoms, it is worth considering a differential diagnosis of HLH. PMID- 11853351 TI - Experienced emotion and affect recognition in adult attention-deficit hyperactivity disorder. AB - Emotional competence and deficits that may disrupt interpersonal interactions were evaluated in 28 adults with attention-deficit hyperactivity disorder (ADHD) and 28 demographically equivalent controls. Participants completed tasks assessing affect recognition and experienced emotional intensity. Adults with ADHD performed worse in affect recognition than did adults without the disorder; however, the impairment was unrelated to gross perceptual processes, fundamental abilities in facial recognition, or attentional aspects of affect perception. Moreover, intensity of experienced emotion moderated affect recognition: Among controls, experienced emotion facilitated affect recognition. Among adults with ADHD, who reported significantly greater intensity, experienced emotion was inversely related to affect recognition. Results are consistent with theories of ADHD as a deficit in behavioral inhibition; yet, results may merely reflect a constellation of deficits associated with the disorder. PMID- 11853352 TI - Perceptually based implicit learning in severe closed-head injury patients. AB - This study suggests that perceptually based implicit learning may either be preserved following a severe closed-head injury (CHI) or recover within the 1st year. Nineteen severe CHI patients and 19 controls searched visual matrices and indicated the quadrant location of a target. Participants were exposed to the following covariation pattern: AAAABAAA. For Covariation A blocks, the matrices systematically co-occurred with a unique location of the target. This relationship was altered for the B block. Despite CHI participants' overall slower response times (RTs), both groups demonstrated the expected decline in RTs across the first 4 Covariation A blocks followed by an increase when the covariation changed. Both groups also exhibited retention of their learning after a 20-min delay. Explicit knowledge tests indicated that participants lacked awareness for the covariation. PMID- 11853353 TI - A prospective study of short- and long-term outcomes after traumatic brain injury in children: behavior and achievement. AB - Longitudinal behavior and achievement outcomes of traumatic brain injury (TBI) were investigated in 53 children with severe TBI, 56 children with moderate TBI, and 80 children with orthopedic injuries not involving brain insult. Measures of preinjury child and family status and of postinjury achievement skills were administered shortly after injury. Assessments were repeated 3 times across a mean follow-up interval of 4 years. Results from mixed model analysis revealed persisting sequelae of TBI. Recovery of math skills was observed in the severe TBI group but only for children from less stressed families. Social disadvantage in children with TBI predicted more adverse behavioral sequelae and less favorable changes in some outcome measures. The findings suggest that pediatric TBI has long-term effects on behavior and achievement but that postinjury progress is influenced by the family environment. PMID- 11853354 TI - Dissociation of sensitivity and response bias in children with attention deficit/hyperactivity disorder during central auditory masking. AB - Forty-three children (ages 7.0-14.5 years old) with and without attention deficit/hyperactivity disorder (ADHD), combined type had thresholds for detection of a 500-Hz pure tone estimated with and without a noise masker in the contralateral ear. The ear receiving the signal in the masked condition was varied randomly. A single-interval maximum-likelihood method estimated thresholds and false-alarm rate. Whereas the increase in threshold in children with ADHD in the presence of contralateral masking was comparable with controls, the increase in false-alarm rate was significantly greater. This dissociation between changes in sensitivity and response bias in the presence of masking noise supports suggestions that children with ADHD have difficulty inhibiting maladaptive responses and indicates that this deficit is quantifiable using psychoacoustic methods. PMID- 11853355 TI - Differential asymmetries for recognizing nouns and verbs: where are they? AB - To support categorical representation in the brain for grammatical class, it is necessary to show that noun-verb differences are attributable to parts of speech and not to covarying semantic factors. Prior visual-half field investigations of noun-verb processing have confounded grammatical class with imageability. The current study included numerous tests of differential noun-verb processing across visual fields for stimuli equated for imageability. Task (lexical decision, pronunciation) and list context (blocked vs. mixed lists) variables were examined in 168 right-handed participants. There was no reliable reduction of the right visual field advantage for moderately imageable nouns as compared with verbs. If there are qualitative hemisphere differences in single-word noun and verb recognition, these may be attributable to semantic dimensions that tend to covary with grammatical class. PMID- 11853356 TI - Neuropsychological characteristics of three subgroups of Flemish delinquent adolescents. AB - Sixty-three adjudicated adolescents, aged 14-17 years, were followed for 2 years to examine the neuropsychological characteristics of subgroups of delinquents. Nonrecidivistic subjects (n = 29) and late recidivistic subjects (n = 22) differed from early recidivistic subjects (n = 12) on intelligence, self-control functioning, and memory. Late recidivistic subjects, compared with not recidivistic subjects, showed a lower verbal IQ. The differences remained when controlling for the level of substance abuse. This study suggests that neuropsychological assessment may help in differentiating delinquent adolescents, although more research is needed to unravel the mediating influences of substance abuse and early neuropsychological and academic problems. PMID- 11853357 TI - Attention, learning, and memory performances and intellectual resources in Vietnam veterans: PTSD and no disorder comparisons. AB - Attention, learning, memory, and estimated intellectual potential were examined in 26 Vietnam veterans diagnosed with posttraumatic stress disorder (PTSD) and in 21 Vietnam veterans without mental disorders. Results revealed PTSD-associated cognitive deficits on tasks of sustained attention, working memory, initial learning, and estimated premorbid intelligence but not on measures of focus of attention, shift of attention, or memory savings. Cognitive task performances adjusted for estimated native intelligence remained negatively correlated with PTSD severity. An intellectual measure adjusted for cognitive task performances was negatively correlated with PTSD severity, even after the authors statistically controlled the level of combat exposure. Results suggested that although intellectual resources may constitute a vulnerability-protective factor for PTSD development, PTSD was associated with cognitive impairment independent of intellectual functioning. PMID- 11853358 TI - Cultural variation in verbal versus spatial neuropsychological function across the life span. AB - Established culture-invariant measures are needed for cross-cultural assessment of verbal and visuospatial speed of processing and working memory across the life span. In this study, 32 younger and 32 older adults from China and from the United States were administered numerically based and spatially based measures of speed of processing and working memory. Chinese superiority on the numerically based tasks was found for younger adults. Age and increasing task demands diminished this cultural effect, as predicted by the framework proposed by D. C. Park, R. Nisbett, and T. Hedden (1999). However, the visuospatial measures of both working memory and speed of processing did not differ cross-culturally for either age group. The authors concluded that these visuospatial measures provide culture-invariant estimates of cognitive processes in East Asian and Western cultures, but that numerically based tasks show evidence of cultural and linguistic biases in performance levels. PMID- 11853360 TI - Neuropsychological impairment among manic, depressed, and mixed-episode inpatients with bipolar disorder. AB - Previous research has demonstrated broad neurobehavioral abnormalities in bipolar affective disorder (cf. G. Cassens, L. Wolfe, & M. Zola, 1990). However, there have been no comparisons of neuropsychological function across patients with manic, depressed, or mixed subtypes. In the present study, 37 manic, 24 mixed episode, and 25 depressed bipolar I inpatients and 34 control subjects were administered a brief battery of neuropsychological tests. The multivariate and univariate effects of participant group on the neuropsychological measures were uniformly significant (p < .05). Planned contrasts revealed that the bipolar participants performed worse than the controls, and few differences existed between the 3 patient groups. Additionally, the bipolar groups were impaired on 50% of the test battery. These abnormalities were unlikely attributable to differences in psychiatric symptomatology, medical illness, comorbid psychiatric diagnoses, or medication status. Findings imply that acute mood disturbance during bipolar disorder yields significant neurobehavioral dysfunction. PMID- 11853359 TI - A profile of neuropsychological deficits in alcoholic women. AB - Neuropsychological deficits, most notable in executive, visuospatial, and functions of gait and balance, are detectable in alcoholic men even after a month of sobriety. Less well established are the severity and profile of persisting deficits in alcoholic women. The authors used an extensive test battery to examine cognitive and motor functions in 43 alcoholic women who were sober, on average, for 3.6 months. Functions most severely affected in alcoholic women involved visuospatial and verbal and nonverbal working memory processes as well as gait and balance. Areas of relative sparing were executive functions, declarative memory, and upper-limb strength and speed. The authors found that lifetime alcohol consumption was related to impairment severity on Block Design (Wechsler Adult Intelligence Scale-Revised, D. Wechsler, 1981) and verbal and nonverbal working memory, suggesting a dose effect of alcohol abuse. The alcohol related deficits in working memory, visuospatial, and balance implicate disruption of prefrontal, superior parietal, and cerebellar brain systems. PMID- 11853361 TI - Intact perceptual and conceptual priming in temporal lobe epilepsy: neuroanatomical and methodological implications. AB - Explicit memory appears to be supported by medical temporal lobe structures, whereas separate neocortical regions may mediate perceptual and conceptual implicit memory. Children and adults with temporal lobe epilepsy (TLE) and matched controls were administered experimental verbal memory tests. Performance on implicit tests--word identification and word generation--was contrasted with explicit recognition and recall. Encoding conditions emphasized either conceptual or perceptual aspects of study words and were crossed with presentation modality. The priming performance of participants with TLE did not differ from controls, but participants with TLE did show deficits on recognition and recall measures. Thus, intact left temporal cortex does not appear to be necessary for normal implicit memory performance, even when conceptual processing is emphasized at study or test. PMID- 11853362 TI - Regulation of glucose transporter 1 expression by gliclazide in rat L6 myoblasts. AB - Sulphonylureas are known to enhance insulin secretion from the pancreas and its sensitivity of the extrapancreatic target organs. In this study, we clarified a direct extrapancreatic effect of the sulphonylureas and of gliclazide, on the glucose transport system in cultured rat L6 myoblasts, which predominantly expressed glucose transporter 1 (GLUT 1). Our results show that gliclazide stimulated 2-deoxy-[3H]-D-glucose (2DG) uptake, 24 h after treatment, in a dose dependent manner, and it also increased GLUT 1 protein synthesis and mRNA expression; 2DG uptake and GLUT 1 protein synthesis induced by gliclazide were completely blocked by protein kinase A (PKA) inhibitors (H89 and rp-cAMP), and gliclazide increased the intracellular cAMP levels 3 to 24 hr after the treatment. These results show that in rat L6 myoblasts, gliclazide stimulates glucose transport activity by the induction of GLUT 1 gene expression through PKA. PMID- 11853363 TI - Glucose kinetics and splanchnic uptake following mixed meal ingestion in cirrhotic-diabetic subjects. AB - Although glucose intolerance and/or overt diabetes are common in cirrhotic subjects, the mechanism(s) that lead to post-prandial hyperglycemia in cirrhosis are not entirely known. To this aim, we measured whole-body rates of glucose appearance (Ra) and of disappearance (Rd) in cirrhotic-diabetic subjects and in controls, before and following a 4-hr administration of a mixed meal. In the post prandial phase, endogenous and dietary glucose Ra, as well as first-pass splanchnic uptake of dietary glucose, were measured using a double (ie oral and intravenous) glucose tracer technique. In the fasting state, the cirrhotic patients were hyperglycemic (12.0 +/- 1.4 vs 4.4 +/- 0.2 mmol/l in controls, p < 0.001), had a higher glucose Ra (17.0 +/- 2.7 vs 10.2 +/- 0.5 micromol x kg(-1) x min(-1), p < 0.05) and a lower clearance rate (1.51 +/- 0.19 vs 2.32 +/- 0.06 ml x kg x min, p < 0.02). Following the meal, plasma glucose increased to greater values (p < 0.002) in the patients (to 16.8 +/- 2 mmol/l, mean values of the last 40 min) than in the controls (to 7.2 +/- 0.4 mmol/l). Insulin increased in both groups but it was 35% lower (p > 0.05) in the patients. Post-prandial total glucose Ra (cirrhotics: 21.3 +/- 2.6; controls: 19.2 +/- 1.4 pmol x kg(-1) x min( 1)), endogenous Ra (cirrhotics: 7.3 +/- 1.5; controls: 7.0 +/- 1.3 micromol x kg( 1) x min(-1)) and first-pass splanchnic uptake of dietary glucose (cirrhotics: 9.8 +/- 2.6; controls: 11.5 +/- 1.6 micromol x kg x min(-1)), were not different between the 2 groups, whereas glucose clearance remained lower (p<0.001) in the patients (1.31 +/- 0.25 ml x kg(-1) x min)-1)) than in the controls (2.72 +/- 0.26). These data demonstrate that, in cirrhotic-diabetic patients, post-pran dial hyperglycemia is not due to a reduced extraction of dietary glucose nor to an increased endogenous production, but rather to a defect in peripheral glucose clearance, secondary to either insulin-resistance and/or relative insulin deficiency. PMID- 11853364 TI - Relationship between cardiac function and insulin resistance in obese patients. AB - Both overweight and insulin resistance predispose to atherosclerosis and cardiovascular diseases, independently of other risk factors. We studied the relationship between insulin resistance and heart function and dimension in 39 patients with different degrees of obesity. Twenty-six women and 13 men with body mass index (BMI) ranging 26.1-41 kg/m2 (mean +/- SD = 33.9 +/- 3.8), without diabetes, hypertension and heart, liver or kidney diseases were studied. Patients were subdivided into 2 groups, 25 with overweight or grade I obesity (group A) and 14 with severe (grade II or III) obesity (group B). Insulin sensitivity was evaluated by the Insulin Tolerance Test (ITT), performed after an overnight fast and K(ITT) was calculated. Echocardiographic measurements were also assessed. Between the two groups no significant difference was observed for either K(ITT) (group A, K(ITT) = 5.47 +/- 1.30; group B, K(ITT) = 4.57 +/- 1.53) or the ejection fraction (EF%) (group A, 71.40 +/- 6.63; group B, 69.86 +/- 7.43). No correlation was observed between BMI and both the EF% and other echocardiographic measurements. In patients with mild obesity (group A) a significant negative correlation between EF% and KITT (r = -0.62,p < 0.001) was observed. In mild obesity, therefore, cardiac function changes occur in relation to the level of insulin resistance but these changes are not related to mass and/or volume changes. The cause(s) of this relationship is not clear, but most likely involves metabolic or endocrine factors. The increased EF% in moderately obese patients that are insulin-resistant may provide an initial compensatory mechanism but may also contribute to a late cardiac damage. PMID- 11853365 TI - Leptin concentrations do not correlate with fat mass nor with metabolic risk factors in morbidly obese females. AB - AIMS: To investigate the determinants of leptinemia in a cohort of morbid obese females compared to those of normal weight and mild-to-moderate obesity, and the relationships between leptin and metabolic derangements associated with obesity. METHODS: Recruited females were: moderately obese [n=44; body mass index (BMI) 25 40 kg/m2], morbidly obese (n=34; BMI > or = 40 kg/m2) and normal weight volunteers (n=12; BMI 19-25 kg/m2). Fat mass assessed by bioelectrical impedance and fat distribution by waist-to-hip ratio (WHR) were determined in all subjects. Biochemical determinations included plasma leptin, lipoprotein profile, fasting insulin and cortisol. RESULTS: Plasma leptin values were significantly increased in morbid obese patients (54.95 +/- 1.8 ng/ml) compared to those moderately obese (30.2 +/- 1.7 ng/ml; p<0.001) and to controls (9.77 +/- 1.4 ng/ml; p<0.001). Fat and age-adjusted leptin values were not different between groups. When subjects with a BMI <40 kg/m2 were considered, plasma leptin was significantly and positively related to anthropometric variables (BMI, percentage body fat and WHR), total cholesterol, LDL-cholesterol, plasma triglycerides, AST, ALT and uric acid; and negatively with HDL-cholesterol. In contrast, when morbidly obese patients were analyzed separately, no relationships were observed between leptin concentrations and BMI, percentage of adiposity or biochemical variables. For obese patients no significant differences were observed in the adjusted leptin values with respect to the presence of diabetes, dyslipidemia or hypertension. CONCLUSIONS: In morbidly obese women, the plasma leptin concentrations, although increased, do not reflect the amount of adipose stores, and as such, factors other than simply adiposity need to be invoked to explain the variation in leptin values. PMID- 11853367 TI - Mitochondrial 3243 BP mutation: a case report. AB - Mitochondrial DNA (mtDNA) gene defects may play a role in the development of maternally inherited diabetes mellitus and deafness (MIDD). Mutation in the mitochondrial gene at position 3243 was recently identified in several pedigrees of diabetes mellitus and deafness. As the mitochondria play an important role in glucose-stimulated insulin secretion in pancreatic beta-cells, we therefore searched for this mutation in a non-consanguineous family with MIDD from Southern Italy. The mitochondrial 3243 bp mutation of the tRNALEU (UUR) gene was identified in one subject with deafness and Type 1B diabetes mellitus and in his mother. The mother was affected by Type 2 diabetes mellitus, deafness and cardiomyopathy. Our study points out the variable phenotypic expression of this mitochondrial mutation. This may suggest the presence of other mitochondrial or nuclear mutations required to modulate the phenotype. PMID- 11853366 TI - Cigarette smoking is a risk factor for nephropathy and its progression in type 2 diabetes mellitus. AB - Cigarette smoking is a risk factor for diabetic nephropathy in Type 1 diabetes (T1DM); a few reports support this possibility in Type 2 diabetes (T2DM) as well. Since heterogeneity among populations could exist, we investigated the association of cigarette smoking and nephropathy, and progression of nephropathy in Italian T2DM patients. A retrospective study was conducted in 273 long duration T2DM subjects with a 3-year follow-up in the out-patient clinic, and at least one access per year. Albumin excretion rate, serum creatinine, and a number of other parameters implicated in the development of diabetic renal disease were evaluated. Progression of nephropathy was defined as the passage from different stages of renal involvement (no renal derangement, microalbuminuria, proteinuric disease or severe nephropathy). At baseline, 13.2% of the subjects had microalbuminuria, and 3.7% proteinuric disease. Microalbuminuria and proteinuric disease were more frequent in actual smokers than in non- and former smokers (chi2=8.35; p=0.015). Progression of nephropathy was less common in non- and former smokers than in smokers (31 of 134, 23%, and 15 of 67, 22%, and 30 of 72, 42%, respectively; chi2=9.32;p=0.009). From logistic regression analysis, smoking (p=0.0012) emerged as the most important factor associated with progression of nephropathy, followed by packyears (p=0.011), HbA1c mean value at follow-up (p=0.024), and total cholesterol (p=0.038). In conclusion, cigarette smoking is a risk factor for progression of nephropathy also in Italian T2DM patients; reducing or quitting smoking should be part of the therapy or of the preventive measures in these patients and their relatives. PMID- 11853368 TI - The potential role of insulin analogues in the treatment of children and adolescents with Type 1 diabetes mellitus. AB - The main therapeutic challenge in the treatment of Type 1 diabetes is maintenance of near-normoglycaemia in order to prevent long-term complications and avoid hypoglycaemia. This goal is relevant from the onset of the disease and is feasible if physiological models of insulin replacement are used and patients are educated in the strategy of intensive insulin therapy. Although the use of available insulins within a multiple injection regimen has improved, metabolic control it is still far from being optimal. The recent introduction of insulin analogues with a short- and long-acting profile seems promising in improving metabolic control and quality of care. Insulin lispro and insulin aspart, the short-acting insulin analogues offer a better post-prandial profile, while insulin glargine the new long-acting insulin analogue might provide better overnight control. In fact, the theoretical combination of an acute prandial insulin peak with a flat interprandial and overnight plasma profile would closely mimic the 24-hr insulin profile of non-diabetic individuals. This would possibly lead to lower post-prandial blood glucose excursion and better fasting blood glucose associated with minimal risk of hypoglycaemia. The possible reduction of hypoglycaemia is especially important in children as recurrent episodes might represent a potential risk for cognitive impairment. However, recent clinical research on the short-acting insulin analogues demonstrates the difficulties of translating these theoretical benefits into clinical relevant advantages. This might happen to other insulin analogues and requires further and larger studies in order to fully exploit the theoretical advantages of insulin analogues in the paediatric population. Safety issues should also be carefully monitored when introducting analogues in long-term therapy. PMID- 11853369 TI - Supporting children through these uncertain times. PMID- 11853370 TI - School-based teen pregnancy prevention programs: a review of the literature. AB - Teenage pregnancy is a well-documented problem in the United States, with approximately 890,000 teenage pregnancies occurring each year. Although teen pregnancy rates have declined since 1991, rates remain higher than the mid-1970s and are fourfold those of European countries. Substantial morbidity and social problems result from these pregnancies, affecting the mother, her children, other family members, and society. Multiple educational approaches have been used, with few demonstrating significant reductions in teen pregnancy. School-based programs have been diverse and multifaceted. Recently, programs with a comprehensive approach have shown potential for success. In this article, characteristics and elements of promising school-based programs are identified and discussed. It is imperative that school nurses play an active role in developing and implementing prevention programs that incorporate rigorous evaluation. As health educators, school nurses are in a prime position to implement and evaluate the effectiveness of teen pregnancy prevention programs. PMID- 11853371 TI - The importance of research to school nurses and school nursing practice. AB - Historically, school nursing has not documented sufficiently the health issues in schools, nor has it prioritized these issues for school nursing interventions or evaluated the effectiveness of nursing interventions. The National Association of School Nurses (NASN) is strongly committed to the advancement of children's health. Thus, NASN is developing an infrastructure to support research that articulates the health conditions affecting children's school success and the contributions of school nursing to children's health and academic success. In 1999, NASN participated with the National Center for School Health and the National Nursing Coalition for School Health at a Research Summit to set a school nursing research agenda. Needs and issues were identified for future research activities. The primary outcome was the identification that school nurses must practice the standards of care pertaining to research. These standards specify that clinicians be informed about, supportive of, and participate in the conduct and use of research. PMID- 11853372 TI - School nurses' experiences, concerns, and knowledge of growth disorders in children: development of a monograph. AB - Growth disorders may be associated with difficult psychosocial adjustment, learning problems, and specific health risks. Appropriate school health programming relies on school nurses who are skilled in growth assessment, management of psychosocial and behavioral problems, and effective communication with school personnel, children, families, and health care resources. A monograph and model individualized healthcare plans were developed for growth disorders in school-age children as an educational resource for school nurses. Knowledge of growth disorders among nurses receiving the monograph was evaluated in a random sample of 336 school nurses, members of the National Association of School Nurses. Knowledge of growth assessment and individualized health care plans for children with specific growth disorders was significantly higher in the group of school nurses who received the monograph. Specific obstacles to implementing school-based health care for children with growth disorders are discussed. PMID- 11853373 TI - School nurses: unsung heroes. PMID- 11853374 TI - Personal and professional characteristics of exemplary school nurses. AB - This article describes a study conducted to identify the professional and personal characteristics of exemplary school nurses. During Phase 1 of the study, a content analysis was completed using nomination portfolios of 10 school nurses who were selected as School Nurse of the Year from 1986 to 1998 in the state of New York. Further content analyses were conducted using transcriptions of a focus group and individual open-ended interviews with exemplary school nurses. These analyses yielded 54 personal characteristics and 74 professional characteristics of exemplary school nurses. Using a Delphi technique in Phase 2 of the study, these characteristics were validated and refined by a national sample of exemplary school nurses who had been chosen to be their state nominees for the National Association of School Nurses' School Nurse of the Year award. Descriptive data were collected on the organizational/structural characteristics of the school systems in which these nurses practiced and on the nurses' individual characteristics. This article identifies areas for clinical and research use of the study's findings. PMID- 11853375 TI - HIV/AIDS peer education: a rural health project. AB - This article describes a program conducted by a group of adolescents in a rural western Nebraska high school that was designed to inform their peers about the risk of HIV/AIDS. The program was funded by state and county agencies. An AIDS Task Force composed of community health leaders developed the guidelines for the program. The Task Force met annually to plan for the coming year, implement changes, and evaluate the program. A community health nurse served as the coordinator of the program and also served as a rural school nurse. A group of students called peer helpers carried out the HIV/AIDS program. Peer helpers created an awareness of HIV/AIDS with dissemination of factual information and also served as a referral resource for their peers. A number of recommendations are provided for future implementation of programs designed to help adolescents develop a healthy lifestyle. PMID- 11853376 TI - Enhancing school nurse visibility. AB - The school nurse is in a powerful position in the school district, but only by expanding visibility efforts can it be a viable one. School nurses are viewed as "health experts" in their school districts. They need to continually fine-tune and expand their visibility, as well as to widen their circle of influence in the school district and its community-based partnerships. Feeling comfortable in giving school board presentations and participating in other highly visible activities will highlight and enhance the valuable services school nurses provide. School nurses must be visible to be perceived as a powerful influence for health in the school district. PMID- 11853377 TI - How to cook up a healthy summer camp. AB - This article is designed to help school nurses plan a children's summer camp. In many ways, it is like a recipe, filled with tried experiences leading to success. Many of the ideas in this article may help others to brainstorm about implementing a camp. The idea for this summer camp originated from an obesity study conducted during the school year. In that study, it was found a large number of students were obese, and the majority of them spent a considerable amount of time in sedentary activities. In addition, the study questionnaire findings indicated that the children were not eating enough fruits or vegetables and had diets with high carbohydrate and fat content. Therefore, the major objective for the summer camp was to introduce the children to structured exercise and nutrition classes. Evaluation indicated the children profited socially, physically, and intellectually from the camp's activities. PMID- 11853378 TI - Preparation of magnetic dye affinity adsorbent and its use in the removal of aluminium ions. AB - Aluminium has recently been considered as a causative agent in dialysis encephalopathy, osteodystrophy, and anemia occuring in hemodialysis patients. The aim of this study is to prepare magnetic poly(2-hydroxyethylmethacrylate) (mPHEMA) adsorbent and to investigate it's useability for the removal of Al(III) ions from drinking and dialysis water. Magnetic PHEMA beads in a size range 80 120 microm were produced by a dispersion polymerization technique. Then Alizarin Red was covalenlty attached onto the mPHEMA beads. Al(III) adsorption from aqueous solutions was examined by batch system. mPHEMA beads were characterized by swelling tests, electron spin resonance (ESR), scanning electron microscopy (SEM), and elemental analysis. Important results obtained in this study are as follows: the swelling ratio of mPHEMA beads was 34%. The presence of magnetite in the polymeric structure was confirmed by ESR. The mPHEMA beads have a spherical shape and porous structure. Alizarin Red loading was 135.8 micromol g(-1) polymer. The maximum Al(III) adsorption was 722 micromol g(-1) polymer at pH 5.0. Non-specific Al(III) adsorption was about 23 micromol g(-1) polymer under the same conditions. High desorption ratios (98%) were achieved by using 0.1 M HNO3. It was possible to reuse the beads without significant loss of Al(III) adsorption capacity. PMID- 11853379 TI - Studies of a novel human thrombomodulin immobilized substrate: surface characterization and anticoagulation activity evaluation. AB - Immobilization of the anticoagulative or antithrombogenic biomolecule has been considered as one of the important methods to improve the blood compatibility of artificial biomaterials. In this study, a novel immobilization reaction scheme was utilized to incorporate the human thrombomodulin, an endothelial cell associated glycoprotein, onto the cover glass surface with an aim to develop an anticoagulative substrate. Trichlorotriazine and amino-terminated silane were employed as the coupling agents, while the polyethylene glycol with a molecular weight of 1500 was used as the spacer in this reaction scheme. Protein C activation assay indicated the immobilized human thrombomodulin still has this coenzymatic activity but is lower, possibly due to the conformation variation by the coupling agents. In vitro platelet adhesion assay has demonstrated the surface with immobilized human thrombomodulin is much less platelet-activating than others. Therefore, the novel reaction scheme proposed here is very promising for future development of an anticoagulative silicon or cover glass substrate (e.g. implantable sensor or biochip) by the immobilization of antithrombogenic protein, such as the human thrombomodulin in this study. PMID- 11853380 TI - Interaction of blood components with heparin-immobilized polyurethanes prepared by plasma glow discharge. AB - The blood compatibility of poly(ethylene oxide) (PEO)-grafted and heparin (Hep) immobilized polyurethanes was investigated using in vitro plasma recalcification time (PRT), activated partial thromboplastin time (APTT), platelet adhesion and activation, and peripheral blood mononuclear cell (PBMC) adhesion and activation. In the experiment with plasma proteins, the PRT of the polyurethane (PU) surface was prolonged by PEO grafting and further prolonged by heparin immobilization. The APTT was prolonged on PU-Hep, suggesting the binding of immobilized heparin to antithrombin III. The percentage of platelet adhesion on PU was not much different from that on acrylic acid- and PEO-grafted PUs (PU-C, PU-6, PU-33), yet was substantially decreased by heparin immobilization (PU-6-Hep, PU-33-Hep). The release of serotonin from adhering platelets was slightly suppressed on PEO grafted PUs yet significantly suppressed on heparin-immobilized PUs. In the PBMC experiments, the adhesion and activation of the cells were significantly suppressed on heparin-immobilized PUs, and the amount of interleukin-6 (IL-6) released from PBMCs stimulated with surface-modified PUs decreased with a decrease in PBMC adhesion. PMID- 11853381 TI - Transience in polyion complexation between nicotinamide-modified dextran and carboxymethyl dextran during enzymatic degradation of dextran. AB - A self-regulated degradation system using polyion complexation through oxidation reaction from degradation products was preliminarily studied. 1,4 Dihydronicotinamide-modified dextran (NAH-Dex) with different molecular weights was prepared, and NAH moiety in NAH-Dex was oxidized by H2O2 to the dehydrated form (NA+-Dex). The dependence of stoichiometry, concentration, and molecular weight on polyion complexation with carboxymethyl dextran (CMD) were examined. NA+-Dex with a molecular weight above 40000 formed an insoluble complex with CMD, and the complexation was found to proceed stoichiometrically. The extent of polyion complexation was dependent on the concentration of NA+-Dex and CMD, whereas the time to reach complexation was dependent on H2O2 concentration. When H2O2 and dextranase were added to the solution containing NAH-Dex, CMD, and dextran, transmittance dropped and then increased again. From these results, the addition of dextran into the system of H2O2, NAH-Dex, CMD, and dextranase can regulate formation and dissociation of the polyion complex between NA+-Dex and CMD. The antagonistical inhibition of the degradation of the polyion complex is a key parameter of the self-regulated degradation system. PMID- 11853382 TI - Selective binding of albumin on stearyl poly(ethylene oxide) coupling polymer modified poly(ether urethane) surfaces. AB - A tri-block-coupling polymer of stearyl poly(ethylene oxide)-4,4'-methylene diphenyl diisocyanate-stearyl poly(ethylene oxide) (MSPEO), was used as a surface modifying additive (SMA) and the MSPEO-modified poly(ether urethane) (PEU) surfaces were prepared by the process of dip-coating. The surface analysis by XPS revealed the surface enrichment of poly(ethylene oxide) (PEO). On the coating modified surfaces, the bovine serum albumin (BSA) adsorption, respectively, from the low and high BSA bulk concentration solutions was correspondingly characterized by the methods of radioactive 125I-probe and ATR-FTIR. The bovine serum fibrinogen (Fg)-adsorption from the Fg bulk solution and the BSA-Fg competing adsorption from the BSA-Fg binary solutions were also characterized by radioactive 125I-probe. The reversible BSA-selective in situ adsorption on MSPEO modified PEU surfaces were achieved, and the performance of blood compatibility on the coating-modified surfaces was also confirmed, respectively, by plasma recalcification time (PRT) and prothrombin time (PT) tests. PMID- 11853383 TI - Fibroblast culture on surface-modified poly(glycolide-co-epsilon-caprolactone) scaffold for soft tissue regeneration. AB - Novel porous matrices made of a copolymer of glycolide (G) and epsilon caprolactone (CL) (51 : 49, Mw 103000) was prepared for tissue engineering using a solvent-casting particulate leaching method. Poly(glycolide-co-epsilon caprolactone) (PGCL) copolymer showed a rubber-like elastic characteristic, in addition to an amorphous property and fast biodegradability. In order to investigate the effect on the fibroblast culture, PGCL scaffolds of varying porosity and pore size, in addition to surface-hydrolysis or collagen coating, were studied. The large pore-sized scaffold (pore size >150 microm) demonstrated a much greater cell adhesion and proliferation than the small pore-sized one. In addition, the higher porosity, the better the cell adhesion and proliferation. The surface-hydrolyzed PGCL scaffold showed enhanced cell adhesion and proliferation compared with the unmodified one. Type I collagen coating revealed a more pronounced contribution for increased cell interactions than the surface hydrolyzed one. These results demonstrate that surface-modified PGCL scaffold can provide a suitable substrate for fibroblast culture, especially in the case of soft tissue regenerations. PMID- 11853384 TI - Immobilization of acetylcholinesterase and choline oxidase in/on pHEMA membrane for biosensor construction. AB - In this study, acetylcholinesterase (AChE) and choline oxidase (ChO) were co immobilized on poly(2-hydroxyethyl methacrylate) (pHEMA) membranes with the aim of using them in biosensor construction. pHEMA membranes were prepared with the addition of different salts in different HEMA: aqueous solution ratios and characterized in terms of porosity, thickness. permeability, and mechanical properties. Membranes prepared in the presence of SnCl4 were found to be superior in terms of porosity and permeability and were chosen as the immobilization matrix. Immobilization of the enzymes was achieved both by entrapment and surface attachment via epichlorohydrin (Epi) and Cibacron Blue F36A (CB) activation. The effect of immobilization on enzyme activity was evaluated by the comparison of Km and Vmax values for the free and immobilized bi-enzyme systems. The increase in Km was negligible (1.08-fold) for the bi-enzyme system upon immobilization on surface but was 2.12-fold upon entrapment. Specific activity of the free enzyme system was found to be 0.306 mV s(-1) microg(-1) ChO while it was 0.069 (4.43 fold decrease) for entrapped and 0.198 (1.54 fold decrease) for CB-Epi immobilized enzymes. The performance of immobilized enzymes in different buffer types, pH, and temperature conditions were evaluated. The best enzyme activity was obtained at pH 9.0. Activity of the enzymes was found to increase with increasing temperature (in the range 25-40 degrees C). PMID- 11853385 TI - Biocompatibility assessment of polytetrafluoroethylene/wollastonite composites using endothelial cells and macrophages. AB - The aim of the study was to prepare a composite of polytetrafluoroethylene/wollastonite (PTFE/W) and evaluate its biocompatibility with endothelial cells. A composite of PTFE with wollastonite in the proportion 90/10 w/w was prepared. The dynamic storage modulus of composite is found to increase from 260 to about 453 MPa at room temperature while a marginal increase is observed in the compressive modulus. Higher values of storage modulus of PTFE/W relative to pristine PTFE over a range of temperature indicated the contribution of wollastonite in improving the rigidity of PTFE. Electron microscopic visualization of composite surface indicates suitable morphology for cell growth with the cross-section showing no evidence of bonding between PTFE and wollastonite. The water contact angle of the composite indicates increased hydrophilicity over native PTFE due to the presence of wollastonite. A direct contact test did not show any deleterious effects on endothelial cell morphology and viability, indicating its compatibility. Leached-out products (LOP) from the composite were determined to be non-toxic as tested by tetrazolium (MTT) and Neutral red uptake (NRU) assays. Mouse peritoneal macrophages cultured in the presence of the composites did not show upregulation of activation markers such as CD11b/CD 18 (Mac-1), CD45, CD 14, and CD86 (B7.2) in comparison to macrophages cultured in contact with PTFE alone, indicating its non-activating nature. LOP did not induce proliferation of mouse splenic lymphocytes suggesting its immuno tolerance. In static incubation assay contact with composite did not lead to hemolysis thus exhibiting preliminary hemocompatibility of the material. Suitable physico-chemical properties and well tolerance by endothelial cells and macrophages make this composite a prospective biomaterial. One could foresee the applications of this composite in areas where materials need to possess high rigidity and are subject to elevated temperatures. PMID- 11853386 TI - Effect of carbomer concentration and degree of neutralization on the mucoadhesive properties of polymer films. AB - Mucoadhesive drug delivery systems may enable a prolonged and localized drug release at various sites of the gastrointestinal tract. In the present study, the mucoadhesive properties of flexible polymeric films based on PVP or PVA as film forming polymers were assessed by measuring the detachment force from excised porcine duodenal mucosa using a tensile strength tester. The mucoadhesive films were comprised of an impermeable backing layer of cellulose acetate butyrate. Carbopol 934P, Carbopol 974NF, and Noveon AA1 were incorporated as mucoadhesive excipients in concentrations of 0-22 wt% relative to the dry mass of the mucoadhesive layer and with various degrees of neutralization corresponding to pH 4.8, 5.5, 6.8, or 7.5. Films based on PVP were generally more mucoadhesive than corresponding formulations based on PVA. Maximum adhesion of PVP-films was observed at pH 5.5 and 6.8 depending on the type of the mucoadhesive polymer and its concentration. An optimal mucoadhesive polymer concentration range was found to be between 2 and 10 wt%. Higher polymer concentrations did not further enhance the mucoadhesive properties, and in some cases even decreased mucoadhesion. Film formulations based on PVA demonstrated no satisfactory mucoadhesive strength. PMID- 11853387 TI - Stability assessment of chitosan-sodium hexametaphosphate capsules. AB - The assessment of the stability of capsules based on chitosan-sodium hexametaphosphate complex formation has been carried out using two independent methods--compression and osmotic swelling, and the influence of the preparation variables was evaluated. The formulation containing 1.5% core polymer (chitosan) and 1.5% oligophosphate, in the absence of salt or at low ionic strength (0.15% NaCl) was found to provide the best membrane resistance. A higher concentration of cross-linker (2.25%) produced stable capsules only in absence of electrolyte. Mannitol, a porogen added to the preparation solutions, did not affect the stability of the obtained membranes. At elevated polyol (1%) and cross-linker levels (2.25%), and at 0% salt, membranes with decreased elasticity were obtained, having lower compression and osmotic bursting values and lower deformation at the breaking points. A significant influence of salt amount on the capsule stability was also found. This was attributed to changes in the membrane formation process resulting in membranes with different thickness and structure. Membrane compression stability was found to be dependent on the pH of both oligophosphate and chitosan solutions, as well as on the reaction time. The bursting force values decreased for capsule diameters below 1.6 mm. The increased membrane/capsule volume ratio for the small capsules decreased the capsule deformation freedom and caused capsule rupture at low force values. The capsules made at low salt amounts showed very good storage stability over time and at elevated temperatures. The results demonstrated that the capsules could be formulated with controlled properties for various biomedical applications. PMID- 11853388 TI - Porous starch-based drug delivery systems processed by a microwave route. AB - Abstract-A new simple processing route to produce starch-based porous materials was developed based on a microwave baking methodology. This innovative processing route was used to obtain non-loaded controls and loaded drug delivery carriers, incorporating a non-steroid anti-inflammatory agent. This bioactive agent was selected as model drug with expectations that the developed methodology might be used for other drugs and growth factors. The prepared systems were characterized by 1H and 13C NMR spectroscopy which allow the study of the interactions between the starch-based materials and the processing components, i.e, the blowing agents. The porosity of the prepared materials was estimated by measuring their apparent density and studied by comparing drug-loaded and non-loaded carriers. The behaviour of the porous structures, while immersed in aqueous media, was studied in terms of swelling and degradation, being intimately related to their porosity. Finally, in vitro drug release studies were performed showing a clear burst effect, followed by a slow controlled release of the drug over several days (up to 10 days). PMID- 11853390 TI - In situ forming lactic acid based orthopaedic biomaterials: influence of oligomer chemistry on osteoblast attachment and function. AB - The ability of osteoblasts to attach and function normally on scaffolds fabricated from synthetic materials is essential for musculoskeletal tissue engineering applications. In this study, the osteoconductivity of polymer networks formed from multifunctional lactic acid oligomers was assessed. These oligomers form highly crosslinked networks via a photoinitiated polymerization, which provides potential advantages for many orthopaedic applications. Depending on the initial oligomer chemistry and the resultant polymer hydrophobicity, protein adsorption and osteoblast function varied significantly between the various lactic acid based polymer chemistries. Results were compared to control polymers of tissue culture polystyrene (TCPS) and 50:50 poly(lactic-co-glycolic acid) (PLGA). The viability of osteoblasts attached to poly(2EG10LA) and poly(2EG6LA) was close to the TCPS and PLGA after 7 and 14 days of culture, whereas cell viability was approximately 50% lower on poly(8EG6LA). Additionally, the alkaline phosphatase activity and mineralization of attached osteoblasts were similar on poly(2EG10LA) and PLGA, whereas these markers of bone formation were significantly lower for poly(2EG6LA) and poly(8EG6LA). For example, the alkaline phosphatase activity of rat calvarial osteoblasts attached to poly(2EG10LA) was 0.048 +/- 0.006 micromol mg(-1) protein-min, but only 0.030 +/- 0.003 micromol mg(-1) protein-min for osteoblasts attached to poly(8EG6LA) after 14 days of culture. Finally, osteoblasts were seeded onto three-dimensional scaffolds to demonstrate the applicability of the scaffolds for bone tissue engineering. PMID- 11853389 TI - Islet immunoisolation: experience with biopolymers. AB - Incidence of Type I diabetes is increasing globally and has become a major health concern. There is enough evidence suggesting involvement of autoimmunity in destruction of insulin-producing islets of langerhans which leads to impaired glucose homeostasis. Islet transplantation is one of the approaches that received wide attention. Due to the autoimmune nature of the disease. strategies to protect transplanted islet graft from rejection are sought. Immunoisolation of islets inside semipermeable biocompatible materials is amongst them. Natural biopolymers have been used extensively as immunoisolation materials due to their satisfactory biocompatiblity and tissue tolerance. Here we attempt to address the need for islet immunoisolation and our experience in using natural biopolymers such as chitosan, cellulose and alginate for this application. PMID- 11853391 TI - Mapping of the distribution of significant proteins and proteoglycans in small intestinal submucosa by fluorescence microscopy. AB - Because small intestine submucosa (SIS) is a bioscaffold for tissue regeneration, we describe a method to analyze the material for growth peptides and for structural molecules. Immunofluorescence methods are described for relative quantification of abundant structural proteins. Additionally, a quantitative technique for comparison of the content of less abundant proteins in SIS was developed using the tyramide signal amplification (TSA) system that is applicable to paraffin-preserved tissue blocks. Frozen sections generally shredded when cut thinly enough to permit entry and washout of reagents. Five micrometer sections cut from paraffin blocks were immunolabeled for collagen, heparan sulfate proteoglycans (HSPG), FGF2, TGFbeta, and VEGF. Images of tissue sections were acquired by a linear image camera and quantified by densitometry after thresholding the signal to minimize nonspecific fluorescence. Immunohistochemistry was used to confirm the immunofluorescence methods. HSPG was widely distributed but concentrated in vessels. FGF2 was distributed diffusely and was associated with fibrous structures. VEGF was distributed mainly around vessels. TGFbeta was barely detectable above background. Collagen fibrils were distinctly present, and with a two-color fluorescence system, the distribution of components relative to collagen can be assessed. The anatomic structure of SIS is likely to play an important role in the regeneration of tissues, and factors in remnant vessels may facilitate penetration of the matrix along these avenues. PMID- 11853392 TI - Advances in model systems for hepatitis C virus research. PMID- 11853393 TI - E1 protein of bovine papillomavirus 1 is not required for the maintenance of viral plasmid DNA replication. AB - Derivatives of bovine papillomavirus 1 (BPV1) with temperature-sensitive and dominant-negative mutation the E1 gene were used to determine the requirement for E1 in the maintenance of viral plasmid DNA replication. The abilities of these mutant BPV1 genomes to replicate as nuclear plasmids were monitored at permissive (32 degrees C) and nonpermissive (37 degrees C) temperatures in mouse C127 cells. We found that the temperature-sensitive E1 mutant BPV1 genomes replicate as nuclear plasmids as efficiently as does wild-type BPV1 in C127 cells after shifting to the nonpermissive temperature. These findings indicate that BPV1 does not require E1 for the maintenance of viral plasmids. PMID- 11853394 TI - Evidence against an important role for infectivity-enhancing antibodies in Ebola virus infections. AB - The neutralizing and enhancing activities of Ebola virus (EBOV)-specific antibodies were tested among four murine antibodies specific to the surface glycoprotein (GP), a recombinant human monoclonal antibody specific to GP, a polyclonal equine IgG, and serum obtained from a convalescent monkey. All but one of these antibodies neutralized EBOV infectivity of primary human monocytes/macrophages or Vero cells. None of the antibodies enhanced EBOV infectivity in these cells. Taken together with in vivo observations that early deaths were not observed in animals immunized with various viral vectors expressing EBOV GP, it is unlikely that any EBOV-enhancing antibodies profoundly affected EBOV pathogenesis. PMID- 11853395 TI - Comparative analysis of the intracellular location of the high- and low-risk human papillomavirus oncoproteins. AB - We have compared the intracellular location of the HPV E6 and E7 proteins from high- and low-risk virus types. While high-risk HPV E7 displays diffuse nuclear expression, low-risk E7 has a nuclear punctuate pattern of expression. Similarly, while high-risk E6 is expressed throughout the cell, low-risk E6 is again predominantly nuclear with a punctuate pattern of expression. Both low-risk viral oncoproteins show colocalization with PML, whereas high-risk viral proteins do not. Finally, inhibition of the proteasome pathway results in a dramatic nuclear accumulation of high-risk E6 protein. These results demonstrate fundamental differences in the localization of these viral oncoproteins within the cell and offer alternative explanations for their respective differences in pathology. PMID- 11853396 TI - Generation of E3-deleted canine adenoviruses expressing canine parvovirus capsid by homologous recombination in bacteria. AB - E3-deleted canine adenovirus type 1 (CAV-1) was generated by homologous recombination in bacterial cells, using an antibiotic resistance marker to facilitate the recovery of recombinants. This marker was flanked by unique restriction endonuclease sites, which allowed its subsequent removal and the insertion of cassettes expressing the canine parvovirus capsid at the E3 locus. Infectious virus was recovered following transfection of canine cells and capsid expression was observed by RT-PCR from one of the virus constructs. A second construct, containing a different promoter, showed delayed growth and genome instability which, based on the size difference between these inserts, suggests a maximum packaging size of 106 to 109% wild-type genome size for CAV-1. PMID- 11853397 TI - Hepatitis C virus nonstructural proteins are localized in a modified endoplasmic reticulum of cells expressing viral subgenomic replicons. AB - For many years our knowledge on hepatitis C virus (HCV) replication has been based on in vitro experiments or transfection studies. Recently, the first reliable system for studying viral replication in tissue culture cells was developed. Taking advantage of this system, we examined in detail the localization of viral nonstructural (NS) proteins in cells containing functional replication complexes. By fractionation experiments and immunomicroscopy, we observed that all NS proteins were associated with the endoplasmic reticulum (ER) membranes, confirming the hypothesis that the ER is the site of membrane associated HCV RNA replication. Interestingly, NS3 and NS4A were preferentially localized in endoplasmic reticulum cisternae surrounding mitochondria, suggesting additional subcellular compartment-related functions for these viral proteins. Furthermore, the immunoelectron microscopy revealed the loss of the organization and other morphological alterations of the ER (convoluted cisternae and paracrystalline structures), resembling alterations observed in liver biopsies of HCV-infected individuals and in flavivirus-infected cells. PMID- 11853399 TI - Chemokine induction and leukocyte trafficking to the lungs during murine gammaherpesvirus 68 (MHV-68) infection. AB - Murine gammaherpesvirus 68 replicates in the alveolar epithelium and induces an inflammatory infiltrate in the lung, following intranasal challenge, and is cleared 10 and 13 days after infection by a T-cell-dependent mechanism. In order to understand the development of the immune response to this virus and how leukocyte trafficking to the lung is regulated, chemokine expression during MHV 68 infection was examined in lung tissue using an RNase protection assay. Expression of RANTES, eotaxin, MIP-1 alpha, MIP-1 beta, IP-10, and MCP-1 was upregulated by day 7 after infection. Chemokine concentrations in lung lavage fluid were also determined by ELISA. MCP-1, RANTES, MIP-1 alpha, eotaxin, and KC were upregulated during MHV-68 infection. Most of these chemokines have been reported to be chemoattractants for either activated T cells or monocytes, which are the major cellular components of the inflammatory infiltrate induced by the virus. Upregulated expression of the corresponding receptors for the chemokines, including CCR1, CCR2, CCR3, CCR5, and CXCR3, coincided with the development of the inflammatory infiltrate. The chemokine levels peaked at around day 7 after infection, coinciding with peak viral titers and slightly preceding maximal T cell infiltration. In vitro chemotaxis assays confirmed that lung lavage fluid from MHV-68-infected mice had chemotactic activity, which was partially blocked by antibodies to IP-10 and RANTES. These observations suggest that the chemokines detected play an important role in regulating leukocyte trafficking to the lungs during MHV-68 infection. PMID- 11853398 TI - Identification of a nucleocapsid protein (VP35) gene of shrimp white spot syndrome virus and characterization of the motif important for targeting VP35 to the nuclei of transfected insect cells. AB - To identify the protein encoded by a 687-bp open reading frame (ORF) of a salI genomic DNA fragment of shrimp white spot syndrome virus (WSSV), we expressed the ORF in a baculovirus/insect cell expression system. The apparent molecular mass of the recombinant protein on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was 35 kDa in insect cells. Antibody raised against bacterially synthesized protein of the ORF identified a nucleocapsid protein (VP35) in the extracts of both the purified WSSV virions and the nucleocapsids which comigrated with the 35-kDa baculovirus-expressed recombinant protein on SDS PAGE. We also show by transient expression in insect cells (Sf9) that VP35 targets the nucleus. Two potential nuclear localization signals (NLSs) were characterized, but only one of them was important for targeting VP35 to the nuclei of transfected insect cells. Replacement of a cluster of four positively charged residues ((24)KRKR(27)) at the N terminus of the protein with AAAA resulted in mutant proteins that were distributed only in the cytoplasm, thus confirming that this sequence is a critical part of the functionally active NLS of VP35. PMID- 11853400 TI - Characterisation of Sri Lankan cassava mosaic virus and Indian cassava mosaic virus: evidence for acquisition of a DNA B component by a monopartite begomovirus. AB - Two bipartite begomoviruses, Indian cassava mosaic virus (ICMV) and Sri Lankan cassava mosaic virus (SLCMV), have been isolated from mosaic-diseased cassava originating from central India and Sri Lanka, respectively. ICMV was transmitted with low efficiency from cassava to Nicotiana benthamiana by sap inoculation to give leaf curl symptoms. SLCMV was much more virulent in this host, producing severe stunting, leaf curl, and chlorosis. These symptoms were reproduced when their cloned genomic components (DNAs A and B) were introduced into N. benthamiana by either mechanical or Agrobacterium-mediated inoculation (agroinoculation). SLCMV is more closely related to ICMV (DNA A, 84%; DNA B, 94% nucleotide identity) than African cassava mosaic virus (ACMV) (DNA A, 74%; DNA B, 47% nucleotide identity). Sequence comparisons suggest that SLCMV DNA B originated from ICMV DNA B by a recombination event involving the SLCMV DNA A intergenic region. Pseudorecombinants produced by reassortment of the cloned components of ICMV and ACMV were not infectious in N. benthamiana, emphasising their status as distinct virus species. In contrast, a pseudorecombinant between ACMV DNA A and SLCMV DNA B was infectious. Consistent with these observations, iteron motifs located within the intergenic region that may be involved in the initiation of viral DNA replication are conserved between SLCMV and ACMV but not ICMV. When introduced into N. benthamiana by agroinoculation, SLCMV DNA A alone produced a severe upward leaf roll symptom, reminiscent of the phenotype associated with some monopartite begomoviruses. Furthermore, coinoculation of SLCMV DNA A and the satellite DNA beta associated with ageratum yellow vein virus (AYVV) produced severe downward leaf curl in N. glutinosa and yellow vein symptoms in Ageratum conyzoides, resembling the phenotypes associated with AYVV DNA A and DNA beta infection in these hosts. Thus, SLCMV DNA A has biological characteristics of a monopartite begomovirus, and the virus probably evolved by acquisition of a DNA B component from ICMV. PMID- 11853401 TI - Structural studies of human enteric adenovirus type 41. AB - Enteric adenoviruses of serotypes 40 and 41 possess some specific structural features, one of which is the presence on the virion of two fibers of different lengths and primary sequences. These viruses are notoriously difficult to grow under laboratory conditions. In this paper the successful growth and purification of Ad41 are presented in detail. Structural Ad41 proteins were analyzed by biochemical methods, mass spectrometry, and electron microscopy (EM), in order to identify and localize them on polyacrylamide denaturing gels and to assess the proportion of short and long fibers in the virion. Surprisingly, the three proteins composing virus short and long pentons did not totally enter the denaturing polyacrylamide gels, which is probably due in part to their high pI. The pentons were separately purified and their dimensions were estimated from EM data. The EM images suggest that there are the same amounts of short and long fibers in each virion. PMID- 11853402 TI - NS1 protein of parvovirus B19 interacts directly with DNA sequences of the p6 promoter and with the cellular transcription factors Sp1/Sp3. AB - The nonstructural proteins of parvovirus exert a variety of disparate functions during viral infection ranging from promoter regulation, involvement in DNA replication, and induction of apoptosis. Our interest was focused on the possible mechanism by which the NS1 protein mediates its effects on the p6 promoter of parvovirus B19. It is known that the p6 promoter is highly active in different cell lines and interaction with the viral NS1 protein results in a further increase of the activity. The protein may function by binding directly to the viral DNA or via an indirect binding through interaction with cellular transcription factors bound to the promoter. We examined the interaction of the NS1 protein with cellular transcription factors which are involved in regulating the promoter activity. After purified baculovirus-expressed NS1 protein in gel retardation assays was added, an altered complex formation was observed, indicating that NS1 protein interacts with Sp1/Sp3 transcription factors. Enzyme linked immunosorbent assays verified these findings. The direct interaction of NS1 protein with p6 promoter elements was analyzed by a coprecipitation assay whereby labeled oligonucleotides spanning the entire promoter region were incubated with NS1 protein followed by an immunoprecipitation with NS1-specific antibodies. An eight-nucleotide-long, almost palindromic sequence (AGGGCGGA) was found as potential NS1-binding motif. Footprint analysis with oligonucleotides containing this DNA motif confirmed this result. Thus, transcriptional regulation by the NS1 protein may involve both the interaction with Sp1/Sp3 that binds to the promoter region and direct binding of NS1 to the promoter DNA. PMID- 11853403 TI - Resting CD4(+) T cells with CD38(+)CD62L(+) produce interleukin-4 which contributes to enhanced replication of T-tropic human immunodeficiency virus type 1. AB - A significant increase in the CD38(+) population among T lymphocytes has been observed in human immunodeficiency virus type 1 (HIV-1)-infected carriers. We previously reported a higher replication rate of T-tropic HIV-1 in the CD4(+)CD38(+)CD62L(+) than CD38(-) subset under conditions of mitogen stimulation after infection. Here, we revealed a similarly high susceptibility in the CD38(+) subset on culture with conditioned medium containing Th2 cytokine, interleukin (IL)-4 that was produced endogenously from this subset on stimulation with mitogen or anti-CD3 antibody for 3 days. The contribution of IL-4 to the upregulated production of virus in the CD38(+) subset was confirmed by culture of this subset with recombinant human IL-4. In contrast, the rate of replication in the CD38(-) subset was not augmented in the conditioned medium from either subset or with IL-4. However, there were no differences in the surface expression of IL 4 receptor or HIV-1 receptors CD4 and CXCR4 between the two subsets. Thus, the CD4(+)CD38(+)CD62L(+) subset comprises a specific cell population secreting endogenous Th2 cytokine that contributes to the efficient production of T-tropic HIV-1 through upregulation at a certain stage of the viral life cycle, probably after the adsorption step. PMID- 11853404 TI - A new cellular factor recognizes E2 binding sites of papillomaviruses which mediate transcriptional repression by E2. AB - Repression of transcription by the full-length E2 protein of papillomaviruses (PV) seems to occur when the E2 binding sites and those of positively acting cellular factors overlap. Previously, we showed that RUNX1 (formerly called CBF) binds to the repression-mediating E2 binding site P2 of human PV type 8 (HPV8). By a yeast one-hybrid system we could identify an unknown protein binding also to P2, tentatively called PBF (papillomavirus binding factor). PBF recognizes the sequence CCGG, which represents the 3' half of the E2 binding site just adjacent to the RUNX1 motif. PBF also binds to the repression-mediating E2 BS-1 in BPV1, which is conserved to P2 of HPV8. Point mutations destroying PBF binding to HPV8 P2 and BPV-1 E2 BS-1 in vitro reduce promoter activity in corresponding reporter constructs. Our results suggest that PBF might play a role in transcription of PV genes and in E2-mediated repression. PMID- 11853405 TI - Characterization of phi 12, a bacteriophage related to phi 6: nucleotide sequence of the small and middle double-stranded RNA. AB - The isolation of additional bacteriophages containing segmented double-stranded RNA genomes has expanded the Cystoviridae family to nine members. Comparing the genomic sequences of these viruses has allowed evaluation of important genetic as well as structural motifs. These comparative studies are resulting in greater understanding of viral evolution and the role played by genetic and structural variation in the assembly mechanisms of the cystoviruses. In this regard, the small and middle double-stranded RNA genomic segments of bacteriophage phi 12 were copied as cDNA and their nucleotide sequences determined. This genome's organization is similar to that of the small and middle segments of bacteriophages phi 6, phi 8, and phi 13. Although there is little similarity in the nucleotide sequences, similarity exists in the amino acid sequence of the lysis cassette proteins to those of phi 6. The host cell attachment proteins are found to have marked similarity to the phi 13 attachment proteins. PMID- 11853406 TI - Complete genome sequence, taxonomic assignment, and comparative analysis of the untranslated regions of the Modoc virus, a flavivirus with no known vector. AB - We recently developed a model for flavivirus infection in mice and hamsters using the Modoc virus (MODV), a flavivirus with no known vector (P. Leyssen, A. Van Lommel, C. Drosten, H. Schmitz, E. De Clercq, and J. Neyts, 2001, Virology 279, 27-37). We now present the coding and noncoding sequence of MODV. The Modoc virus genome was determined to be 10,600 nucleotides in length with a single open reading frame extending from nucleotides 110 to 10,234, encoding 3374 amino acids. The deduced gene order of the single open reading frame is C-prM-E-NS1 NS2A-NS2B-NS3-NS4A-NS4B-NS5, which is exactly the same as that of the mosquito- and tick-borne flaviviruses. It is flanked by a 5'- and 3'-untranslated region (UTR) of 109 and 366 nucleotides, respectively. Alignment of the MODV amino acid sequence with that of 20 other flaviviruses revealed several regions with high sequence similarity corresponding to functionally important domains (e.g., the serine protease/helicase/NTPase of NS3 and the methyltransferase/RNA-dependent RNA polymerase of NS5) and conserved sites for proteolytic cleavage by viral and cellular proteases. Phylogenetic analysis of the entire coding region confirmed the classification of MODV within the flaviviruses with no known vector, which is in agreement with previous findings based on partial NS5 sequences. A detailed comparative analysis of the putative folding patterns of the 5'- and 3'-UTR of MODV and of the tick- and mosquito-borne viruses was carried out. Structural elements in the 5'- and 3' UTR of MODV that are preserved among vector-borne flaviviruses were noted and so were structural elements distinguishing the MODV UTRs from mosquito-borne and tick-borne flaviviruses. Also the putative secondary structure of circularized MODV RNA is presented. PMID- 11853407 TI - Autogenous translational inhibition of core protein: implication for switch from translation to RNA replication in hepatitis C virus. AB - Positive-stranded viruses use the genomic RNA as a common template for translation and RNA replication which proceed in inverse direction; a certain regulatory mechanism for translation control is probably required to coordinate these two antagonistic processes. Hepatitis C virus (HCV) core protein is a good candidate that might play a role in such a regulation. In this study, we further investigated whether HCV core protein modulates internal ribosome entry site (IRES)-directed translation. The inclusion of the core-coding sequence significantly suppressed translation initiated by HCV IRES in monocistronic and bicistronic reporter systems. The region mainly responsible for this inhibition was mapped to nt 441-473 of the core-coding sequence. This suppression was eliminated by frameshift mutations introduced into this region, suggesting that it is the core protein expressed in cis, rather than the core-coding nucleotide sequence that negatively modulates the efficiency of HCV IRES-dependent translation. Furthermore, the core protein provided in trans also specifically decreased the IRES activity in directing cap-independent translation both in transfected cells and in cell-free translation study. Consistently, a gel mobility shift assay showed a specific interaction between the core protein and HCV IRES-containing RNA transcript. These findings suggest that HCV core protein may down-regulate the cap-independent translation as a regulatory mechanism required for initiation of transcription. PMID- 11853408 TI - Human cytotoxic T lymphocyte responses to live attenuated 17D yellow fever vaccine: identification of HLA-B35-restricted CTL epitopes on nonstructural proteins NS1, NS2b, NS3, and the structural protein E. AB - Yellow fever virus (YFV) is a re-emerging problem despite the existence of an effective live-attenuated vaccine. The induction of YFV-neutralizing antibodies undoubtedly contributes to vaccine efficacy, but T lymphocyte responses to YFV likely play a role in long-term efficacy. We studied the T lymphocyte responses to YFV in four vaccinees. Proliferation and cytolytic responses to YFV were demonstrated in all subjects. We isolated 13 YFV-specific CD8(+) CTL lines that recognized epitopes on the E, NS1, NS2b, and NS3 proteins; eight CTL lines were HLA-B35-restricted. YFV-specific T cell responses were detectable by IFN gamma ELISPOT assays 14 days postvaccination, with T cell frequencies sustained for up to 19 months. To our knowledge, this is the first report of human T lymphocyte responses following YFV vaccination. These results indicate that the live 17D YFV vaccine induced CD8(+) T cell responses directed against at least four different HLA-B35-restricted YFV epitopes. PMID- 11853409 TI - Rapid activation of poly(ADP-ribose) polymerase contributes to Sindbis virus and staurosporine-induced apoptotic cell death. AB - Poly(ADP-ribose) polymerase-1 (PARP-1) is a chromatin-associated enzyme that is activated by DNA strand breaks and catalyzes the transfer of ADP-ribose groups from NAD to itself and other nuclear proteins. Although caspase-mediated PARP-1 cleavage occurs during almost all forms of apoptosis, the contribution of PARP-1 activation and cleavage to this cell death process remains unclear. Using immortalized fibroblasts from wild-type (PARP-1(+/+)) and PARP-1 knockout (PARP 1(-/-)) mice, and a mouse neuroblastoma cell line (N18), the role that poly(ADP ribosyl)ation plays in Sindbis virus (SV)-induced apoptosis was examined. Robust PARP-1 activation occurred in SV-infected cells prior to morphologic changes associated with apoptotic cell death and PARP-1 activity ceased simultaneously with caspase-3 activation and PARP-1 proteolysis. PARP-1 activity was maximal before detectable DNA fragmentation, but was absent when DNA damage was most intense. SV and staurosporine-induced cell death was delayed in fibroblasts lacking PARP-1 activity, suggesting that PARP-1 activation contributes to apoptotic cell death induced by these stimuli. SV replication was not affected by lack of PARP-1 activity, but DNA fragmentation and caspase-3 activation were delayed and occurred at lower levels in PARP-1-deficient fibroblasts. Early virus induced PARP-1 activation may represent a novel way by which cells signal to the nucleus to regulate protein function by poly(ADP-ribosyl)ation in response to virus infection. PMID- 11853410 TI - Adenoassociated virus type 2-induced inhibition of the human papillomavirus type 18 promoter in transgenic mice. AB - The epithelium of the cervix uteri has been reported to be frequently coinfected with both human papillomaviruses (HPV) and helper virus-dependent adenoassociated viruses (AAV). Seroepidemiological data suggest that AAV infection could inhibit cervical cancer that is caused by specific ("high-risk") types of papillomaviruses. In vitro, infection with AAV type 2 (AAV-2) or transfection of AAV-2 early (rep) genes has been shown to inhibit transformation by papillomaviruses. To analyze the effects of AAV on HPV in vivo, we studied the influence of AAV-2 infection on the promoter activity of high-risk HPV type 18 (HPV-18) in mice, transgenic for sequences of the upstream regulatory region (URR) of HPV-18 controlling transcription of the reporter gene, lacZ. Transgenic animals (or tongue cells thereof, explanted and grown in culture) were treated with dexamethasone to induce the HPV-18 promoter. Simultaneously they were (i) infected with AAV, (ii) inoculated with AAV virus-like particles (VLPs; empty capsids), or (iii) mock infected. Inoculation with AAV-2 or VLPs inhibited activation of the HPV-18 promoter. In vitro, in baby hamster kidney cells transfected with the HPV-18-lacZ construct, tissue extracts from AAV-infected animals suppressed the HPV-18 URR to a similar extent as AAV infection did. Down regulation of the HPV-18 promoter was less efficient with extracts from animals inoculated with VLPs and was not observed with extracts from uninfected or dexamethasone-treated animals. This indicates that AAV induces cellular factor(s) in vivo capable of mediating down-regulation of the HPV-18 promoter also in cells in vitro. In contrast, promoters of the low-risk HPV types (HPV-6, HPV-11) were not influenced by AAV infection as opposed to promoters of the high-risk types (HPV-18 and HPV-16). PMID- 11853411 TI - Uptake and processing of modified bacteriophage M13 in mice: implications for phage display. AB - Internalization and degradation of filamentous bacteriophage M13 by a specific target cell may have major consequences for the recovery of phage in in vivo biopanning of phage libraries. Therefore, we investigated the pharmacokinetics and processing of native and receptor-targeted phage in mice. (35)S-radiolabeled M13 was chemically modified by conjugation of either galactose (lacM13) or succinic acid groups (sucM13) to the coat protein of the phage to stimulate uptake by galactose recognizing hepatic receptors and scavenger receptors, respectively. Receptor-mediated endocytosis of modified phage reduced the plasma half-life of native M13 (t(1/2) = 4.5 h) to 18 min for lactosylated and 1.5 min for succinylated bacterophage. Internalization of sucM13 was complete within 30 min after injection and resulted in up to 5000-fold reduction of bioactive phage within 90 min. In conclusion, these data provide information on the in vivo behavior of wild-type and receptor-targeted M13, which has important implications for future in vivo phage display experiments and for the potential use of M13 as a viral gene delivery vehicle. PMID- 11853412 TI - Controlled cell killing by a recombinant nonsegmented negative-strand RNA virus. AB - In most tissue culture cell lines tested, infection with the paramyxovirus simian virus 5 (SV5) results in very little cell death. To determine if SV5 could be used as a vector for controlled killing of tumor cells, a recombinant SV5 (rSV5 TK) was constructed to encode the herpes simplex virus thymidine kinase (TK) gene. MDBK cells infected with rSV5-TK showed a time-dependent loss of viability when infected cells were cultured in the presence of the prodrug acyclovir (ACV) or ganciclovir (GCV) while no significant toxicity was observed in the absence of prodrug. Cells infected with a control rSV5 expressing GFP and cultured with prodrug showed only a slight reduction in growth rate and little cell death. Time lapse video microscopy of rSV5-TK-infected MDBK cells that were cultured in the presence of ACV showed an accumulation of cells with morphological effects characteristic of apoptotic cell death. An MDBK cell line persistently infected with rSV5-TK retained long-term expression of TK and sensitivity to prodrug mediated cell killing that were comparable to those found in an acute infection. Titration experiments showed that the rSV5-TK plus GCV combination resulted in cell death for all mouse and human cell lines tested, although the kinetics and efficiency of cell death varied between cell types. Our results demonstrating controlled cell killing by a recombinant paramyxovirus support the use of negative-strand RNA viruses as therapeutic vectors for targeted killing of cancer cells. PMID- 11853413 TI - Dramatic 5'-residue effect on conformer distribution of short oligonucleotide retro models of the cisplatin-DNA cross-link: implications for the Lippard and cross-link distorted base pair steps present in cisplatin-DNA duplex adducts. AB - The cisplatin anticancer drug preferentially attacks the GG sequence of DNA duplexes. Virtually all DNAs containing the key G*G* lesion (G* = N7 platinated G) have large distortions in the cross-link (G*G*) base pair (bp) step and also in the adjacent Lippard (XG*) bp step, making the adducts very different from B form DNA in the XG*G* region. The XG*G* strand in duplexes also differs in several ways from single-strand (ss) models with G*G* and XG*G* sequences. In the duplex, the X residue has an N sugar, the 5'-G* and 3'-G* bases have slight "R" canting (3'-G* H8 atom toward the 5'-G* base), and there is no or weak H-bonding by the NH3 ligands. In most XG*G* ss models, X has an S sugar, the 5'-G* base normally cants strongly toward the 3'-G* base (L canting), and the NH3 forms an H bond. Well-defined ss models exist in the solid state, but dynamic motion obscures the properties of the ss models in solution. In this work, we employ retro models (better defined, less dynamic ss models) to understand the differences between duplex and ss models. The retro models in this study lack carrier ligand NH's, thus eliminating H-bonding. To correlate previous ss solid state models with our solution work, we constructed hybrid molecules by overlaying parts of known structures. The combined model and experimental information indicates that the X N-pucker is not favorable in L-canted ss models, that X residue steric effects (not H-bonding) favor L canting in ss models, that X N-pucker is needed for favorable WC hydrogen bonding and stacking interactions in duplexes, and that X N-pucker minimizes X base clashes with bases in the complementary strand in duplexes. The R canting minimizing clashes between the X and G* residues of the Lippard bp step (independent of X pucker) and the repositioning of the X residue base caused by the change from S-pucker to N pucker together lead to the unusual features of the Lippard bp step in the duplex. PMID- 11853414 TI - Moving beyond molecules: patterning solid-state features via dip-pen nanolithography with sol-based inks. AB - Herein, we described a new dip-pen nanolithography (DPN)-based method for the direct patterning of organic/inorganic composite nanostructures on silicon and oxidized silicon substrates. The approach works by the hydrolysis of metal precursors in the meniscus between an AFM tip and a surface according to the reaction 2MCln + nH2O --> M2On + 2nHCl; M = Al, Si, and Sn. The inks are hybrid composites of inorganic salts with amphiphilic block copolymer surfactants. Three proof-of-concept systems involving Al2O3, SiO2, and SnO2 nanostructures on silicon and silicon oxide surfaces have been studied. Arrays of dots and lines can be written easily with control over feature size and shape on the sub-200 nm level. The structures have been characterized by atomic force microscopy, scanning electron microscopy, transmission electron microscopy, and energy dispersive X-ray analysis. This work is important because it opens up the opportunity for using DPN to deposit solid-state materials rather than simple organic molecules onto surfaces with the resolution of an AFM without the need for a driving force other than chemisorption (e.g., applied fields). PMID- 11853415 TI - Infrared spectra of gas-phase V(+)-(benzene) and V(+)-(benzene)(2) complexes. AB - The organometallic ions V+-(benzene) and V+-(benzene)2 are produced by laser vaporization in a pulsed nozzle source. They are trapped and mass selected in an ion-trap/time-of-flight mass spectrometer, and their infrared spectra are measured with resonance-enhanced multiphoton photodissociation (IR-REMPD) spectroscopy with a tunable free-electron laser. Vibrational bands in the 600 1800 cm-1 region are characteristic of the benzene molecular moiety perturbed by the metal cation bonding. Experimental data are compared to the IR spectra derived from density functional calculations. Vibrational patterns in V+-(C6H6) indicate that the metal is bound in an eta6 pi-bonding configuration, while V+ (C6H6)2 is a sandwich. Trapped-ion IR-REMPD is a general method to access the vibrational spectroscopy of organometallic ions and their clusters. PMID- 11853416 TI - Copper-based bioinspired oxygenation and glyoxalase-like reactivity. AB - Re-engineered, structurally abbreviated models of metalloenzymes may extend their biomimetic functionality to bioinspired reactivity. The oxygenation of external substrates, in particular, remains an important objective of biomimetic and bioinspired catalysis. We report that the reaction of [(Cu(I)TpCF3,CH3)2] with excess acetone in air produces [CuTpCF3,CH3)(lactate)] in over 95% yield at ambient conditions, without any noticeable ligand decomposition. This chemically unprecedented one-pot conversion of acetone to lactate occurs as a multistep process in the gluconeogenic pathway catalyzed by P450 isozyme 3a and Ni- or Zn based glyoxalases. On the basis of the structure of the [CuTpCF3,CH3)(lactate)] product and oxygenation experiments using isotopically labeled acetone and water, an inner-sphere oxidation/isomerization mechanism is proposed. PMID- 11853417 TI - Influence of regio- and stereoregularity of propene insertion on crystallization behavior and elasticity of ethene-propene copolymers. AB - Melting enthalpies of several ethene-propene samples, obtained by different single-center catalytic systems and having widely different constitutions and compositions in the range of industrial interest, are compared. For the first time, a clear evidence of influence of the regio- and stereoregularity of propene insertion on crystallization behavior of ethene/propene copolymers is provided. This influence is in qualitative agreement with the results of simplified energy calculations relative to models of pseudo-hexagonal packing of ethene-propene copolymers. PMID- 11853418 TI - Redox-active metal-containing nucleotides: synthesis, tunability, and enzymatic incorporation into DNA. AB - Novel redox-active DNA labeling tags with tunable electrochemical potentials are modularly synthesized using (a) bis-substituted Ru2+ or Os2+ precursors (R2bpy)2ML2, (b) substituted 2,4-pentanediones or hydroxamic acids bearing a functionalized linker, and (c) modified nucleotides. DNA polymerase efficiently incorporates the metal-containing nucleotide triphosphate into DNA oligonucleotides. PMID- 11853419 TI - A theoretical study of beta-sheet models: is the formation of hydrogen-bond networks cooperative? AB - The cooperativity in terms of enthalpy contribution for beta-sheet formation of polyglycine models in a vacuum has been studied theoretically by using a repeating unit approach. No cooperativity is found in the parallel direction for both the parallel and antiparallel beta-sheets. Cooperativity in the perpendicular direction is dependent upon the residue number (m) in each beta strand. While there is large cooperativity in the acetamide hydrogen-bond chain (m = 0), the cooperativity is not large in beta-sheet networks (m > 0). SCIPCM solvent model calculations also significantly reduce the cooperativity in hydrogen-bond chains. It is concluded that cooperativity is mainly due to long range electrostatic interactions and not due to the resonance effect. PMID- 11853420 TI - Two unusual, competitive mechanisms for (2-ethynylphenyl)triazene cyclization: pseudocoarctate versus pericyclic reactivity. AB - The cyclization of (2-ethynylphenyl)triazenes in ODCB at 200 degrees C gives exclusively cinnolines, whereas addition of CuCl to 1,2-dichloroethane solutions of the triazenes at 50 degrees C results in the sole formation of isoindazoles. DFT calculations and deuterium labeling studies suggest the intermediacy of a 3 dehydrocinnolinium ion, produced through a pericyclic transition state. Calculations and trapping studies strongly implicate a carbene intermediate in isoindazole formation, which proceeds through a pseudocoarctate transition state. PMID- 11853421 TI - Pentacyclic laddersiloxane. AB - A novel method which effects ring extension of the ladder oligosilsesquioxanes was developed. By applying this method, we synthesized the first functionalized tricyclic laddersiloxanes and pentacyclic laddersiloxane. The reaction of (i PrSi(OH)O)4 with (i-PrPhClSiO)2 in pyridine gave the tetraphenyl tricyclic laddersiloxane. Following dephenylchlorination with AlCl3/HCl, and hydrolysis enabled us to isolate the tetrahydroxyl tricyclic laddersiloxane in good yield. We repeated the similar reaction from this tetrahydroxyl laddersiloxane, and the first pentacyclic laddersiloxane was obtained. The structures of the tetraphenyl and tetrahydroxyl tricyclic laddersiloxanes, and pentacyclic laddersiloxane were determined by X-ray crystallography. PMID- 11853422 TI - Self-assembled monolayers of alkanethiolates on palladium are good etch resists. AB - This paper describes microcontact printing (muCP) of long-chain alkanethiolates on palladium, followed by solution-phase etching with an iron(III)-based etchant, to make patterned structures. The commonly used soft-lithographic procedure for fabricating microstructures-muCP of SAMs on gold-has three shortcomings: a significant surface density of pinhole defects, substantial edge roughness, and incompatibility with processes used in CMOS fabrication. Microcontact printing on palladium gives fewer defects and smaller edge roughness than on gold, and is compatible with CMOS. The mechanism by which etch-resistant patterns are formed is different for palladium and gold. The Pd/S interfacial layer formed by the reaction of palladium films with sulfur-containing compounds provides good resistance to etches independently of the barrier to access the surface provided by the film of (CH2)n groups in the long-chain SAMs. This barrier is the basis of the etch resistance of SAMs on gold, but only supplements the etch resistance of the sulfur-containing interfacial layer on palladium. Characterization of the SAM formed from hexadecanethiol on palladium is described. PMID- 11853423 TI - Enantioselective Staudinger synthesis of beta-lactams catalyzed by a planar chiral nucleophile. AB - The development of efficient methods for the stereoselective generation of beta lactams is an important goal, due to their biological activity and their utility as synthetic intermediates. The Staudinger reaction, an overall [2 + 2] cycloaddition of a ketene with an imine, provides a nicely convergent route to this family of compounds. Nearly all studies to date of asymmetric variants of the Staudinger reaction have focused on the use of chiral auxiliaries to control the stereochemistry of the beta-lactam. In this report, we establish that a planar-chiral derivative of 4-(pyrrolidino)pyridine serves as a very effective enantioselective catalyst for the Staudinger beta-lactam synthesis, coupling a range of symmetrical and unsymmetrical ketenes with an array of imines with very good stereoselection and yield. PMID- 11853424 TI - The metathesis-facilitated synthesis of terminal ruthenium carbide complexes: a unique carbon atom transfer reaction. AB - Ruthenium benzylidene metathesis catalysts react with 2,3-dicarbomethoxymethylene cyclopropane, eliminating styrene and dimethyl fumarate, and producing the first terminal ruthenium carbide complexes. The products are diamagnetic, air-stable, and moderately soluble in hydrocarbon solvents. An X-ray study of Ru(C:)Cl2(P(C6H11)3) (1,3-dimesityl-4,5-dihydroimidazol-2-ylidene) shows a Ru-C distance of 1.650(2) A, consistent with the presence of a very short Ru-C triple bond. PMID- 11853425 TI - A smart photochromophore through synergistic coupling of photochromic subunits. AB - The novel bis-naphthopyran 1 displays optical properties that are unique, differing substantively from a simple first-order superimposition of the monomeric constituents. Such a photochromophore has great promise for the development of smart optical devices. PMID- 11853426 TI - Asymmetric synthesis of 1-aryl-1,2-ethanediols from arylacetylenes by palladium catalyzed asymmetric hydrosilylation as a key step. AB - Double hydrosilylation of arylacetylenes with trichlorosilane catalyzed first by platinum and second by a chiral monophosphine-palladium complex generated the corresponding 1,2-bis(silyl)-1-arylethanes, the oxidation of which with hydrogen peroxide gave 1-aryl-1,2-diols of high enantiomeric purity (94-98% ee) in high yields. PMID- 11853427 TI - Selective Pd-catalyzed oxidative coupling of anilides with olefins through C-H bond activation at room temperature. AB - Using a high-throughput experimentation approach we found a selective and mild Pd catalyzed oxidative coupling reaction between anilide derivatives and acrylates that occurs through ortho C-H bond activation. The reaction is carried out in an acidic environment and occurs even at room temperature with use of a cheap oxidant (benzoquinone) in yields up to 91%. The benzoquinone possibly also functions as a ligand, stabilizing the catalyst. From the electronic dependence of the reaction and the observed kinetic isotope effect (kH/kD = 3) the key step of the catalytic cycle is believed to be electrophilic attack by a [PdOAc]+ complex on the pi-system of the arene. PMID- 11853428 TI - Spatially resolved spectroscopy and structurally encoded imaging by magnetic resonance force microscopy of quadrupolar spin systems. AB - Enhancement of the sensitivity of magnetic resonance force microscopy allowed the extension of the technique to observe half-integer quadrupolar nuclei. This is demonstrated for various different compounds and nuclei with different spin numbers. The possibility of obtaining spatially localized spectral information through the quadrupole interaction is implemented by using nutation NMR. This enables us to superimpose a contrast function on the image of materials depending on their local lattice structure. This opens up new possibilities for both surface and subsurface studies in materials chemistry. PMID- 11853429 TI - Development of a highly efficient catalytic method for synthesis of vinyl ethers. AB - A new method for the preparation of alkyl vinyl ethers has been developed. Thus, various types of alkyl vinyl ethers were synthesized by the reaction of alcohols with vinyl acetate under the influence of a catalytic amount of [Ir(cod)Cl]2 combined with Na2CO3 in good to excellent yields. PMID- 11853430 TI - Mesostructured forms of gamma-Al(2)O(3). AB - gamma-Al2O3 is one of the most extensively utilized metal oxides in heterogeneous catalysis. Conventional forms of this oxide typically exhibit a surface area and pore volume less than 250 m2/g and 0.5 cm3/g, respectively. Previous efforts to prepare mesostructured forms of alumina resulted only in structurally unstable derivatives with amorphous framework walls. The present work reports mesostructured aluminas with walls made of gamma-Al2O3, denoted MSU-gamma. These materials are structurally stable and provide surface areas and pore volumes up to 370 m2/g and 1.5 cm3/g, respectively. The key to obtaining these structures is the formation of a mesostructured surfactant/boehmite precursor, denoted MSU-S/B, assembled through the hydrolysis of an aluminum cation, oligomer, or molecule in the presence of a nonionic surfactant. Mesostructured, gamma-aluminas offer the possibility of improving the catalytic efficiency of many heterogeneous catalytic processes, such as petroleum refining, petrochemical processing, and automobile exhaust control. PMID- 11853431 TI - A combinatorial scaffold approach toward kinase-directed heterocycle libraries. AB - A novel strategy for efficient synthesis of various substituted heterocycles as kinase-directed combinatorial libraries is described. The general scheme involves capture of various dichloroheterocycles onto solid support and further elaborations by aromatic substitution with amines at elevated temperature or by anilines, boronic acids, and phenols via palladium-catalyzed cross-coupling reactions, thus the scaffold itself is transformed into a diversity element within the combinatorial scheme. Libraries consisting of discrete and highly diverse heterocyclic small molecules constructed with these chemistries are currently being evaluated in a variety of cell and protein-based assays. PMID- 11853432 TI - Novel superstructure of nondiscoid mesogens: uneven-parallel association of half disk molecules, 3,4,5-trialkoxybenzoic anhydrides, to a columnar structure and its one-directionally geared interdigitation. AB - Compact and simple nondiscoid mesogens, trialkoxybenzoic anhydrides 2-11 ((RO)(3)C(6)H(2)-CO-O-CO-C(6)H(2)(OR)(3), R = C(2)H(5), n-C(3)H(7), n-C(4)H(9), n C(6)H(13), n-C(8)H(17), n-C(10)H(21), n-C(12)H(25), n-C(14)H(29), n-C(16)H(33), and (S)- and (R)-3,7-dimethyloctyl) were designed and synthesized, and their superstructures were investigated by polarized light microscopy, differential scanning calorimetry, calculation (MM2 and AM1), circular dichroism spectroscopy, and X-ray diffraction. As the result, in the cases of 5-9 and 11, the half-disk molecules in the liquid crystal phases were self-assembled by the dipole-dipole interaction between their carbonyl groups to a column in which the molecules were piled up in an alternately antiparallel manner, and the columns were interdigitated each other. The observed interdigitations were highly directional in the perpendicular direction to the column axis. In the case of compound 11 which has (S)- or (R)-3,7-dimethyloctyl groups, a helical organization of the molecules in the column was estimated from the peaks in the circular dichroism spectra. This is the first report of a one-directionally interdigitated columnar phase, and these compounds are the first liquid crystalline acid anhydrides. It was demonstrated that a -CO-O-CO- moiety is useful as a polar junction in liquid crystalline compounds. PMID- 11853434 TI - Control of multivalent interactions by binding epitope density. AB - Receptor clustering by multivalent ligands can activate signaling pathways. In principle, multivalent ligand features can control clustering and the downstream signals that result, but the influence of ligand structure on these processes is incompletely understood. Using a series of synthetic polymers that vary systematically, we studied the influence of multivalent ligand binding epitope density on the clustering of a model receptor, concanavalin A (Con A). We analyze three aspects of receptor clustering: the stoichiometry of the complex, rate of cluster formation, and receptor proximity. Our experiments reveal that the density of binding sites on a multivalent ligand strongly influences each of these parameters. In general, high binding epitope density results in greater numbers of receptors bound per polymer, faster rates of clustering, and reduced inter-receptor distances. Ligands with low binding epitope density, however, are the most efficient on a binding epitope basis. Our results provide insight into the design of ligands for controlling receptor-receptor interactions and can be used to illuminate mechanisms by which natural multivalent displays function. PMID- 11853433 TI - Ketopremithramycins and ketomithramycins, four new aureolic acid-type compounds obtained upon inactivation of two genes involved in the biosynthesis of the deoxysugar moieties of the antitumor drug mithramycin by Streptomyces argillaceus, reveal novel insights into post-PKS tailoring steps of the mithramycin biosynthetic pathway. AB - Mithramycin is an aureolic acid-type antimicrobial and antitumor agent produced by Streptomyces argillaceus. Modifying post-polyketide synthase (PKS) tailoring enzymes involved in the production of mithramycin is an effective way of gaining further information regarding the late steps of its biosynthetic pathway. In addition, new "unnatural" natural products of the aureolic acid-type class are likely to be produced. The role of two such post-PKS tailoring enzymes, encoded by mtmC and mtmTIII, was investigated, and four novel aureolic acid class drugs, two premithramycin-type molecules and two mithramycin derivatives, were isolated from mutant strains constructed by insertional gene inactivation of either of these two genes. From data bank comparisons, the corresponding proteins MtmC and MtmTIII were believed to act as a C-methyltransferase involved in the production of the D-mycarose (sugar E) of mithramycin and as a ketoreductase seemingly involved in the biosynthesis of the mithramycin aglycon, respectively. However, gene inactivation and analysis of the accumulated products revealed that both genes encode enzymes participating in the biosynthesis of the D-mycarose building block. Furthermore, the inactivation of MtmC seems to affect the ketoreductase responsible for 4-ketoreduction of sugar C, a D-olivose. Instead of obtaining premithramycin and mithramycin derivatives with a modified E-sugar upon inactivation of mtmC, compounds were obtained that completely lack the E-sugar moiety and that possess an unexpected 4-ketosugar moiety instead of the D-olivose at the beginning of the lower deoxysaccharide chain. The inactivation of mtmTIII led to the accumulation of 4E-ketomithramycin, showing that this ketoreductase is responsible for the 4-ketoreduction of the D-mycarose moiety. The new compounds of the mutant strains, 4A-ketopremithramycin A2, 4A-keto-9-demethylpremithramycin A2, 4C-keto-demycarosylmithramycin, and 4E-ketomithramycin, indicate surprising substrate flexibility of post-PKS enzymes of the mithramycin biosynthetic pathway. Although the glycosyltransferase responsible for the attachment of D mycarose cannot transfer the unmethylated sugar to the existing lower disaccharide chain, it can transfer the 4-ketoform of sugar E. In addition, the glycosyltransferase MtmGIV, which is responsible for the linkage of sugar C, is also able to transfer an activated 4-ketosugar. The oxygenase MtmOIV, normally responsible for the oxidative cleavage of the tetracyclic premithramycin B into the tricyclic immediate precursor of mithramycin, can act on a substrate analogue with a modified or even incomplete trisaccharide chain. The same is true for glycosyltransferases MtmGI and MtmGII, both of which partake in the formation and attachment of the A-B disaccharide in mithramycin. PMID- 11853435 TI - Design of a conformationally restricted analogue of the antiepilepsy drug Vigabatrin that directs its mechanism of inactivation of gamma-aminobutyric acid aminotransferase. AB - The antiepilepsy drug vigabatrin (1, 4-aminohex-5-enoic acid, gamma-vinylGABA) is known to be a mechanism-based inactivator of the pyridoxal phosphate (PLP) dependent enzyme gamma-aminobutyric acid aminotransferase (GABA-AT). Inactivation has been shown to proceed by two divergent mechanisms (Nanavati, S. M.; Silverman, R. B. J. Am. Chem. Soc. 1991, 113, 9341-9349). The major pathway involves gamma-proton removal, tautomerization into the PLP ring, followed by Michael addition of an active site lysine residue at the conjugated vinyl group to give a stable covalent adduct with the protein (Scheme 2, pathway a). The minor inactivation mechanism also involves gamma-proton removal, but tautomerization occurs through the vinyl group, followed by an enamine rearrangement that leads to attachment of the inactivator to the PLP, which is bound to the protein (Scheme 2, pathway b). The cause for the two different inactivation pathways was hypothesized to be potential overlap of the incipient carbanion with the pi-orbitals of both the PLP and the vinyl group. With use of the crystal structure data for GABA-AT recently reported (Storici, P.; Capitani, C.; De Biase, D.; Moser, M.; John, R. A.; Jansonius, J. N.; Schirmer, T. Biochemistry 1999, 38, 8628-8634) a computer model of vigabatrin bound to the PLP was constructed and energy minimized. This model indicated that the major Michael addition pathway could only occur if the vinyl group were allowed to rotate by 180 degrees. A conformationally rigid analogue of vigabatrin, cis-3-aminocyclohex 4-ene-1-carboxylic acid (9), was designed to prevent bond rotation and block the Michael addition pathway. A detailed study of the mechanism of inactivation of GABA-AT by 9 revealed that it inactivates by a single mechanism, the enamine pathway. PMID- 11853436 TI - Site-specific probing of oxidative reactivity and telomerase function using 7,8 dihydro-8-oxoguanine in telomeric DNA. AB - Telomeres at the ends of human chromosomes contain the repeating sequence 5' d[(TTAGGG)(n)]-3'. Oxidative damage of guanine in DNAs that contain telomeric and nontelomeric sequence generates 7,8-dihydro-8-oxoguanine (8OG) preferentially in the telomeric segment, because GGG sequences are more reactive in duplex DNA. We have developed a general strategy for probing site-specific oxidation reactivity in diverse biological structures through substitution of minimally modified building blocks that are more reactive than the parent residue, but preserve the parent structure. In this study, 8OG was substituted for guanine at G(8), G(9), G(14), or G(15) in the human telomeric oligonucleotide 5'-d[AGGGTTAG(8)G(9)GTT AG(14)G(15)GTTAGGGTGT]-3'. Replacement of G by 8OG in telomeric DNA can affect the formation of intramolecular G quadruplexes, depending on the position of substitution. When 8OG was incorporated in the 5'-position of a GGG triplet, G quadruplex formation was observed; however, substitution of 8OG in the middle of a GGG triplet produced multiple structures. A clear correspondence between structure and reactivity was observed when oligonucleotides containing 8OG in the 5'-position of a GGG triplet were prepared in the quadruplex or duplex forms and interrogated by mediated electrocatalytic oxidation with Os(bpy)(3)(2+) (bpy = 2,2'-bipyridine). The rate constant for one-electron oxidation of a single 8OG in the 5'-position of a GGG triplet was (6.2 +/- 1.7) x 10(4) M(-1) s(-1) in the G quadruplex form. The rate constant was 2-fold lower for the same telomeric sequence in the duplex form ((3.0 +/- 1.3) x 10(4) M(-1) s(-1)). The position of 8OG in the GGG triplet affects telomerase activity and synthesis of telomeric repeat products. Telomerase activity was decreased significantly when 8OG was substituted in the 5'-position of the GGG triplet, but not when 8OG was substituted in the middle of the triplet. Thus, biological oxidation of G to 8OG in telomeres has the potential to modulate telomerase activity. Further, small molecules that inhibit telomerase by stabilizing telomeric G quadruplexes may not be as effective under oxidative stress. PMID- 11853437 TI - Efficient sialyltransferase inhibitors based on glycosides of N acetylglucosamine. AB - D-Glucosamine was transformed into phenyl and 2-benzoyloxyethyl N acetylglucosamine beta-glycosides 6a and 6b, respectively. Transformation of 6a,b into 6-O-unprotected N-acetylglucosamine derivatives 9a,b permitted the generation of an aldehyde group in the 6-position. Treatment of these intermediates with base afforded unsaturated aldehyde derivatives 10a,b, which are structural mimics of 2,3-dehydroneuraminic acid. H-Phosphonate addition to the aldehyde group and attachment of the cytidine monophosphate residue to the generated hydroxy group gave fully protected transition state analogues of cytidine monophosphate-N-acetylneuraminic acid 14a,b. Liberation of the unprotected compounds 1ah,l and 1bh,l led to excellent inhibitors of alpha(2-6) sialyltransferase from rat liver. Variation of the protective group cleavage procedure for 14a,b led to formal loss of phosphate, thus resulting in diene derivatives (E)-/(Z)-2a,b, which also exhibited inhibitory properties. PMID- 11853438 TI - Investigation of the mechanism of n-butane oxidation on vanadium phosphorus oxide catalysts: evidence from isotopic labeling studies. AB - The selective oxidation of n-butane to maleic acid catalyzed by vanadium phosphates (VPO) is one of the most complex partial oxidation reactions used in industry today. Numerous reaction mechanisms have been proposed in the literature, many of which have butenes, butadiene, and furan as reaction intermediates. We have developed an experimental protocol to study the mechanism of this reaction in which (13)C-isotopically labeled n-butane is flowed over a catalyst bed and the reaction products are analyzed using (13)C NMR spectroscopy. This protocol approximates the conditions found in an industrial reactor without requiring an exorbitant amount of isotopically labeled material. When [1,4 (13)C]n-butane reacted on VPO catalysts to produce maleic acid and butadiene, the isotopic labels were observed in both the 1,4 and 2,3 positions of butadiene and maleic acid. The ratio of label scrambling was typically 1:20 for the 2,3:1,4 positions in maleic acid. For butadiene, the ratio of label scrambling was consistently much higher, at 2:3 for the 2,3:1,4 positions. Because of the discrepancy in the amount of label scrambling between maleic acid and butadiene, butadiene is unlikely to be the primary reaction intermediate for the conversion of n-butane to maleic anhydride under typical industrial conditions. Ethylene was always observed as a side product for n-butane oxidation on VPO catalysts. Fully (13)C-labeled butane produced about 5-13 times as much isotopically labeled ethylene as did [1,4-(13)C]butane, indicating that ethylene was produced mainly from the two methylene carbons of n-butane. When the reaction was run under conditions which minimize total oxidation products such as CO and CO(2), the amounts of ethylene and carbon oxides produced from fully (13)C-labeled butane were almost equal. This strongly suggests that the total oxidation of n-butane on VPO catalysts involves the oxidation and abstraction of the two methyl groups of n-butane, and the two methylene groups of n-butane form ethylene. An organometallic mechanism is proposed to explain these results. PMID- 11853439 TI - Design and synthesis of an enzyme-cleavable sensor molecule for phosphodiesterase activity based on fluorescence resonance energy transfer. AB - Ratiometric measurement is a technique that can provide precise data and even quantitative detection. To carry out ratiometric measurements, it is necessary that the sensor molecule exhibits a large shift in its emission or excitation spectrum after reaction with the target molecule. Fluorescence resonance energy transfer (FRET) is one mechanism used to obtain a large spectral shift. In this study, our aim was to develop a ratiometric fluorescent sensor molecule for phosphodiesterase activity based on FRET. We designed and synthesized CPF4 with a coumarin donor, a fluorescein acceptor, and two phenyl linkers having the phosphodiester moiety interposed between them. In the emission spectrum of CPF4 in aqueous buffer excited at 370 nm, the emission of the coumarin donor was strongly quenched and the emission of the fluorescein acceptor was observed. This emission spectrum demonstrates that energy transfer from the coumarin donor to the fluorescein acceptor proceeds efficiently. Addition of a phosphodiesterase to an aqueous solution of CPF4 resulted in an increase in the donor fluorescence and a decrease in the acceptor fluorescence. CPF4 exhibited a large shift in its emission spectrum after the hydrolysis of the phosphodiester group by the enzyme. This large shift of the emission spectrum indicates that ratiometric measurements can be made by using CPF4. The method described in this paper for designing enzyme-cleavable sensor molecules based on FRET should be readily applicable to other hydrolytic enzymes. PMID- 11853440 TI - The formation of polymer vesicles or "peptosomes" by polybutadiene-block-poly(L glutamate)s in dilute aqueous solution. AB - Polybutadiene-block-poly(L-glutamate) copolymers were made by anionic polymerization and subsequent ring-opening polymerization of N-carboxyanhydrides and were characterized by NMR, IR, SEC, and circular dichroism. These polymers, when appropriately designed, form so-called "polymersomes" or "peptosomes", vesicles composed of modified protein units. The size and structure of the vesicles are determined by dynamic light scattering, small-angle neutron scattering, and freeze-fracture electron microscopy. It is also shown that the size of the peptosomes does not depend on the pH; that is, the solvating peptide units can perform a helix-coil transition without serious changes of the vesicle morphology. PMID- 11853441 TI - Stereocontrolled synthesis of (-)-macrolactin A. AB - The total synthesis of (-)-macrolactin A, a 24-membered macrolide, has been achieved using a newly developed 1,3-diol synthon for the introduction of two key stereogenic centers. The synthon was derived from sequential use of the Noyori asymmetric reduction followed by chiral sulfoxide methodology. Tellurium-derived cuprate organometallics offered an efficient and highly stereoselective means for installation of the C8 Z/E-diene, while the C15 E/E-segment was derived from a Julia-Lythgoe olefination. Yamaguchi lactonization was used to secure the macrocycle in a convergent approach with the longest linear sequence of 19 steps from Noyori alcohol 6. PMID- 11853442 TI - Substituent effects in carbon-nitrogen cleavage of thiamin derivatives. Fragmentation pathways and enzymic avoidance of cofactor destruction. AB - The combination of thiamin and benzaldehyde can produce benzoin but also destroys thiamin. The destruction comes from fragmentation of the conjugate of thiamin and benzaldehyde undergoing a process that produces a phenyl thiazole ketone and pyrimidine. The key step in this process is cleavage of the C-N bond between the heterocycles, which occurs by an unknown mechanism. Enzymes that utilize similar intermediates do not fragment the cofactor although fragmentation is inherent to the structure. To analyze the nature of the C-N cleavage step, the rates of fragmentation of a series of phenyl-substituted N1'-methyl-2-(1 hydroxybenzyl)thiamin derivatives were determined under two sets of conditions: (1) where proton transfer in the step prior to C-N bond breaking is rate determining and (2) where C-N bond breaking is rate-determining. The resulting rho values are 1.6 and 1.8, respectively, leading to the conclusion that C-N cleavage is insensitive to substituent effects. On the basis of these results, we conclude that cleavage occurs by a facile process that resembles the outcome of a [1,5] sigmatropic rearrangement. An enzyme may avoid the fragmentation by holding the intermediate in a conformation that prevents such a process, allowing the normal catalytic process to proceed. PMID- 11853443 TI - Direct observation of aldehyde insertion into rhodium-aryl and -alkoxide complexes. AB - Several organorhodium(I) complexes of the general formula (PPh(3))(2)(CO)RhR (R = p-tolyl, o-tolyl, Me) were isolated and were shown to insert aryl aldehydes into the aryl-rhodium(I) bond. Under nonaqueous conditions, these reactions provided ketones in good yield. The stability of the arylrhodium(I) complexes allowed these reactions to be run also in mixtures of THF and water. In this solvent system, diarylmethanols were generated exclusively. Mechanistic studies support the formation of ketone and diarylmethanol by insertion of aldehyde into the rhodium-aryl bond and subsequent beta-hydride elimination or hydrolysis to form diaryl ketone or diarylmethanol products. Kinetic isotope effects and the formation of diarylmethanols in THF/water mixtures are inconsistent with oxidative addition of the acyl carbon-hydrogen bond and reductive elimination to form ketone. Moreover, the intermediate rhodium diarylmethoxide formed from insertion of aldehyde was observed directly during the reaction. Its structure was confirmed by independent synthesis. This complex undergoes beta-hydrogen elimination to form a ketone. This alkoxide also reacts with a second aldehyde to form esters by insertion and subsequent beta-hydrogen elimination. Thus, reactions of arylrhodium complexes with an excess of aldehyde formed esters by a double insertion and beta-hydrogen elimination sequence. PMID- 11853445 TI - Re(2)(CO)(10)-promoted S-binding, C-S bond cleavage, and hydrogenation of benzothiophenes: organometallic models for the hydrodesulfurization of thiophenes. AB - In hydrodesulfurization model reactions of dinuclear metal complexes with thiophenes, we observe that ultraviolet photolysis of Re(2)(CO)(10) and benzothiophenes (BT) in hexanes solution produces the ring-opened BT complexes Re(2)(CO)(7)(mu-BT) (1a-d) (BT = benzothiophene (BT) 1a, 2-methylbenzothiophene (2-MeBT) 1b, 3-methylbenzothiophene (3-MeBT) 1c, and 3,5-dimethylbenzothiophene (3,5-Me(2)BT) 1d). The eta(1)(S)-bound BT complexes Re(2)(CO)(9)(eta(1)(S)-BT) (2a-d), prepared from Re(2)(CO)(9)(THF) and BT, are readily converted into 1a-d in good yields (40-60%) during UV photolysis in hexanes solution, which suggests that the eta(1)(S)-bound complexes 2a-d are precursors to 1a-d in the reactions of Re(2)(CO)(10) with BT. Irradiation of Re(2)(CO)(10) and 3,5-Me(2)BT with UV light in decane solution under an atmosphere of H(2) produces complex 1d and the partially hydrogenated BT complex Re(2)(CO)(7)(mu-3,5-Me(2)BT-H)(eta-H) (3d). Reactions of 1a with phosphines yield further ring-opened BT-Re complexes of the types Re(2)(CO)(7)(PMe(3))(3)(mu-BT) (4) and Re(2)(CO)(7)(PR(3))(2)(mu-BT) (R = Me (5), (i)Pr (6), Cy (7), and bis(diethylphosphino)ethane (8)). Structures of 1d, 2c, 3d, and 6, which demonstrate various bonding modes of benzothiophene and its C-S cleaved derivatives to two metal centers, were determined by X-ray crystallographic studies. PMID- 11853444 TI - Six-coordinate and five-coordinate Fe(II)(CN)(2)(CO)(x) thiolate complexes (x = 1, 2): synthetic advances for iron sites of [NiFe] hydrogenases. AB - The dicyanodicarbonyliron(II) thiolate complexes trans,cis-[(CN)(2)(CO)(2)Fe(S,S C-R)](-) (R = OEt (2), N(Et)(2) (3)) were prepared by the reaction of [Na][S-C(S) R] and [Fe(CN)(2)(CO)(3)(Br)](-) (1). Complex 1 was obtained from oxidative addition of cyanogen bromide to [Fe(CN)(CO)(4)](-). In a similar fashion, reaction of complex 1 with [Na][S,O-C(5)H(4)N], and [Na][S,N-C(5)H(4)] produced the six-coordinate trans,cis-[(CN)(2)(CO)(2)Fe(S,O-C(5)H(4)N)](-) (6) and trans,cis-[(CN)(2)(CO)(2)Fe(S,N-C(5)H(4))](-) (7) individually. Photolysis of tetrahydrofuran (THF) solution of complexes 2, 3, and 7 under CO led to formation of the coordinatively unsaturated iron(II) dicyanocarbonyl thiolate compounds [(CN)(2)(CO)Fe(S,S-C-R)](-) (R = OEt (4), N(Et)(2) (5)) and [(CN)(2)(CO)Fe(S,N C(5)H(4))](-) (8), respectively. The IR v(CN) stretching frequencies and patterns of complexes 4, 5, and 8 have unambiguously identified two CN(-) ligands occupying cis positions. In addition, density functional theory calculations suggest that the architecture of five-coordinate complexes 4, 5, and 8 with a vacant site trans to the CO ligand and two CN(-) ligands occupying cis positions serves as a conformational preference. Complexes 2, 3, and 7 were reobtained when the THF solution of complexes 4, 5, and 8 were exposed to CO atmosphere at 25 degrees C individually. Obviously, CO ligand can be reversibly bound to the Fe(II) site in these model compounds. Isotopic shift experiments demonstrated the lability of carbonyl ligands of complexes 2, 3, 4, 5, 7, and 8. Complexes [(CN)(2)(CO)Fe(S,S-C-R)](-) and NiA/NiC states [NiFe] hydrogenases from D. gigas exhibit a similar one-band pattern in the v(CO) region and two-band pattern in the v(CN) region individually, but in different positions, which may be accounted for by the distinct electronic effects between [S,S-C-R](-) and cysteine ligands. Also, the facile formations of five-coordinate complexes 4, 5, and 8 imply that the strong sigma-donor, weak pi-acceptor CN(-) ligands play a key role in creating/stabilizing five-coordinate iron(II) [(CN)(2)(CO)Fe(S,S-C-R)](-) complexes with a vacant coordination site trans to the CO ligand. PMID- 11853446 TI - Reaction of nitrogen monoxide with a macrocyclic superoxorhodium(III) complex produces an observable nitratorhodium intermediate. AB - The rapid (k > or = 10(6) M(-1) s(-1)) reaction between NO and L(2)(H(2)O)RhOO(2+) (L(2) = meso-Me(6)-[14]ane-N(4)) generates two strongly oxidizing, scavengeable intermediates, believed to be NO(2) and L(2)(H(2)O)RhO(2+). A mechanism is proposed whereby a peroxynitrito complex L(2)(H(2)O)RhOONO(2+) is formed first. The homolysis of O-O bond produces NO(2) and L(2)(H(2)O)RhO(2+) which were trapped with ABTS(2)(-) and Ni([14]aneN(4))(2+). In the absence of scavengers, the decomposition of L(2)(H(2)O)RhOONO(2+) produces both free NO(3)(-) and a rhodium nitrato complex L(2)(H(2)O)RhONO(2)(2+), which releases NO(3)(-) in an inverse acid-dependent process. The total yield of L(2)(H(2)O)RhONO(2)(2+) is 70%. In a minor, parallel path, NO and L(2)(H(2)O)RhOO(2+) react to give nitrite and the hydroperoxo complex L(2)(H(2)O)RhOOH(2+). PMID- 11853447 TI - Sandia octahedral molecular sieves (SOMS): structural and property effects of charge-balancing the M(IV)-substituted (M = Ti, Zr) Niobate framework. AB - Sandia octahedral molecular sieves (SOMS) is an isostructural, variable composition class of ion exchangers with the general formula Na(2)Nb(2 x)M(IV)(x)O (6-x)(OH)(x).H(2)O (M(IV) = Ti, Zr; x = 0.04-0.40) where up to 20% of the framework Nb(V) can be substituted with Ti(IV) or Zr(IV). This class of molecular sieves is easily converted to perovskite through low-temperature heat treatment (500-600 degrees C). This report provides a detailed account of how the charge imbalance of this Nb(V)-M(IV) substitution is compensated. X-ray powder diffraction with Rietveld refinement, infrared spectroscopy, thermogravimetric analysis, (23)Na MAS NMR, and (1)H MAS NMR were used to determine how the framework anionic charge is cation-balanced over a range of framework compositions. All spectroscopic evidence indicated a proton addition for each M(IV) substitution. Evidences for variable proton content included (1) increasing OH observed by (1)H MAS NMR with increasing M(IV) substitution, (2) increased infrared band broadening indicating increased H-bonding with increasing M(IV) substitution, (3) increased TGA weight loss (due to increased OH content) with increasing M(IV) substitution, (4) no variance in population on the sodium sites (indicated by Rietveld refinement) with variable composition, and (5) no change in the (23)Na MAS NMR spectra with variable composition. Also observed by infrared spectroscopy and (23)Na MAS NMR was increased disorder on the Nb(V)/M(IV) framework sites with increasing M(IV) substitution, evidenced by broadening of these spectral features. These spectroscopic studies, along with ion exchange experiments, also revealed the effect of the Nb(V)/M(IV) framework substitution on materials properties. Namely, the temperature of conversion to NaNb(1-x)M(IV)(x)O(3) (M = Ti, Zr) perovskite increased with increasing Ti in the framework and decreased with increasing Zr in the framework. This suggested that Ti stabilizes the SOMS framework and Zr destabilizes the SOMS framework. Finally, comparing ion exchange properties of a SOMS material with minimal (2%) Ti to a SOMS material with maximum (20%) Ti revealed the divalent cation selectivity of these materials which was reported previously is a function of the M(IV) substitution in the framework. A thorough investigation of this class of SOMS materials has revealed the importance of understanding the influence of heterovalent substitutions in microporous frameworks on material properties. PMID- 11853448 TI - Bimetallic reactivity. One-site addition two-metal oxidation reaction of dioxygen with a bimetallic dicobalt(II) complex bearing five- and six-coordinate sites. AB - The di-Co(2+) complex, [Co(2+)(mu-OH)(oxapyme)Co(2+)(H(2)O)](+), contains an unsymmetrical binucleating ligand (oxapyme) which provides five- and six coordinate metal sites when a hydroxide bridge is introduced. This complex absorbs 1 equiv of O(2) irreversibly in solution, producing an unstable di-Co(3+) oxygenated product. The oxygenated product has been studied at low temperatures, where its electronic absorption and (1)H NMR spectra were recorded. It is probable that the oxygenation reaction involves a one-site addition two-metal oxidation reaction to produce an end-on-bonded peroxide ligand at the available coordination site, giving the complex [Co(3+)(mu-OH)(oxapyme)Co(3+)(mu(1) O(2))](+). Addition of 1 equiv of HClO(4) to this oxygenation product gives a stable peroxide complex, [Co(3+)(mu,eta(1):eta(2)-O(2))(oxapyme)Co(3+)](2+), where one of the oxygen atoms bridges the two metals and is sideways bonded to one of the metals. The formation of this stable complex involves expulsion of the OH(-) bridge. Addition of NO(2)(-) to the sideways-bonded peroxide complex leads to the formation of another stable complex, [Co(3+)(mu,eta(1):eta(1) O(2))(oxapyme)Co(3+)(NO(2))](+), where the peroxide forms a classic di-end-on bridge to the two metals. Both of these complexes have been fully characterized. Addition of acid to this second stable dioxygen complex leads to the release of HNO(2) and the formation of the mu,eta(1):eta(2) sideways-bonded peroxide complex. PMID- 11853450 TI - Carbene formation in its lower singlet state from photoexcited 3H-diazirine or diazomethane. A combined CASPT2 and ab initio direct dynamics trajectory study. AB - The potential energy surfaces of the ground and valence excited states of both 3H diazirine and diazomethane have been studied computationally by mean of the CASSCF method in conjunction with the cc-pVTZ basis set. The energies of the critical points found on such surfaces have been recomputed at the CASPT2/cc-pVTZ level. Additionally, ab initio direct dynamic trajectory calculations have been carried out on the S(1) and S(2) surfaces, starting each trajectory run at the region dominated by the conformational molecular rearrangement of diazomethane. It is found that both isomers are interconnected along a C(s)() reaction coordinate on each potential surface. Radiationless deactivation of the corresponding S(1) state of each isomer occurs through the same point on the surface, an S(1)/S(0) conical intersection. Thereafter, the system has enough energy to surmount the barrier which leads to dissociation products (CH(2) + N(2)) on S(0) state. Therefore, photoexcitation to S(1) state of either diazirine of diazomethane produces methylene in its lower singlet state on a very short time scale (ca. 100 fs). Furthermore, both isomers can generate excited singlet carbene when they are excited onto the S(2) surface; in this case, they lose the activation energy passing through another common S(2)/S(1) conical intersection and then proceed to dissociation into carbene and N(2) on the S(1) surface. For the special case of methylene, it rapidly experiences deexcitation to S(0) state. PMID- 11853449 TI - A supramolecular microfluidic optical chemosensor. AB - A supramolecular microfluidic optical chemosensor (muFOC) has been fabricated. A serpentine channel has been patterned with a sol-gel film that incorporates a cyclodextrin supramolecule modified with a Tb(3+) macrocycle. Bright emission from the Tb(3+) ion is observed upon exposure of the (mu)FOC to biphenyl in aqueous solution. The signal transduction mechanism was elucidated by undertaking steady-state and time-resolved spectroscopic measurements directly on the optical chemosensor patterned within the microfluidic network. The presence of biphenyl in the cyclodextrin receptor site triggers Tb(3+) emission by an absorption energy transfer-emission process. These results demonstrate that the intricate signal transduction mechanisms of supramolecular optical chemosensors are successfully preserved in microfluidic environments. PMID- 11853451 TI - Coherent effects in energy transport in model dendritic structures investigated by ultrafast fluorescence anisotropy spectroscopy. AB - Measurements of ultrafast fluorescence anisotropy decay in model branched dendritic molecules of different symmetry are reported. These molecules contain the fundamental branching center units of larger dendrimer macromolecules with either three (C(3))- or four (T(d), tetrahedral)-fold symmetry. The anisotropy for a tetrahedral system is found to decay on a subpicosecond time scale (880 fs). This decay can be qualitatively explained by Forster-type incoherent energy migration between chromophores. Alternatively, for a nitrogen-centered trimer system, the fluorescence anisotropy decay time (35 fs) is found to be much shorter than that of the tetramers, and the decay cannot be attributed to an incoherent hopping mechanism. In this case, a coherent interchromophore energy transport mechanism should be considered. The mechanism of the ultrafast energy migration process in the branched systems is interpreted by use of a phenomenological quantum mechanical model, which examines the two extreme cases of incoherent and coherent interactions. PMID- 11853452 TI - Exchange interactions in the three phases of vanadyl pyrophosphate (VO)(2)P(2)O(7). AB - The magnetic exchange constants of vanadyl pyrophosphate (VO)(2)P(2)O(7) have been calculated on the basis of a combined DFT/broken symmetry approach. The three reported phases, ambient-pressure orthorhombic, ambient-pressure monoclinic, and high-pressure orthorhombic, have been explicitly considered. Calculations have been performed on four types of model clusters extracted from the crystal lattices. The singularity of each phase is clearly reflected through the number and values of exchange parameters. Our results show that the exchange interactions can be described in first approximation within the alternating antiferromagnetic chain model. The largest exchange coupling along the chain occurs through O-P-O bridges. The interchain interactions are much weaker and are of ferromagnetic nature. PMID- 11853453 TI - Modeling the active site of cytochrome oxidase: synthesis and characterization of a cross-linked histidine-phenol. AB - A cross-linked histidine-phenol compound was synthesized as a chemical analogue of the active site of cytochrome c oxidase. The structure of the cross-linked compound (compound 1) was verified by IR, (1)H and (13)C NMR, mass spectrometry, and single-crystal X-ray analysis. Spectrophotometric titrations indicated that the pK(a) of the phenolic proton on compound 1 (8.34) was lower than the pK(a) of tyrosine (10.1) or of p-cresol (10.2). This decrease in pK(a) is consistent with the hypothesis that a cross-linked histidine-tyrosine may facilitate proton delivery to the binuclear site in cytochrome c oxidase. Time-resolved optical absorption spectra of compound 1 at room temperature, generated by excitation at 266 nm in the presence and absence of dioxygen, indicated a species with absorption maxima at approximately 330 and approximately 500 nm, which we assign to the phenoxyl radical of compound 1. The electron paramagnetic resonance (EPR) spectra of compound 1, obtained after UV photolysis, confirmed the generation of a paramagnetic species at low temperature. Because the cross-linked compound lacks beta-methylene protons, the EPR line shape was dramatically altered when compared to that of the tyrosyl radical. However, simulation of the EPR line shape and measurement of the isotropic g value was consistent with a small coupling to the imidazole nitrogen and with little spin density perturbation in the phenoxyl ring. The ground-state Fourier transform infrared (FT-IR) spectrum of compound 1 showed that addition of the imidazole ring perturbs the frequency of the tyrosine ring stretching vibrations. The difference FT-IR spectrum, associated with the oxidation of the cross-linked compound, detected significant perturbations of the phenoxyl radical vibrational bands. We postulate that phenol oxidation produces a small delocalization of spin density onto the imidazole nitrogen of compound 1, which may explain its unique optical spectral properties. PMID- 11853454 TI - Solvent dependence of the 2-naphthyl(carbomethoxy)carbene singlet-triplet energy gap. AB - The solvent dependence of the 2-naphthyl(carbomethoxy)carbene (2) singlet-triplet energy gap has been examined by time-resolved infrared (TRIR) and computational methods. The ground state of 2 changes from the triplet state in hexane to the singlet state in acetonitrile. Preferential stabilization of the singlet carbene is the result of its increased dipole moment in polar solvents. Variable temperature TRIR experiments provide measurements of the enthalpic and entropic differences between (1)2 and (3)2 and suggest that solvent and geometry effects on the entropy of singlet and triplet carbenes can offset differences arising from spin multiplicity. B3LYP calculations using the polarizable continuum solvation model (PCM) reproduce the general trends in enthalpic differences seen experimentally. PMID- 11853455 TI - A solid-state (17)O nuclear magnetic resonance study of nucleic acid bases. AB - We report a systematic solid-state (17)O NMR study of free nucleic acid bases: thymine (T), uracil (U), cytosine (C), and guanine (G). Site-specifically (17)O enriched samples were synthesized:[2-(17)O]thymine (1), [4-(17)O]thymine (2), [2 (17)O]uracil (3), [4-(17)O]uracil (4), [2-(17)O]cytosine (5), and [6 (17)O]guanine monohydrate (6). Magic-angle-spinning (MAS) and static (17)O NMR spectra were acquired at 11.75 T for compounds 1-6, from which information about the (17)O chemical shift and electric field gradient tensors was obtained. Extensive quantum chemical calculations were performed at the B3LYP/6-311++G(d,p) level of theory for (17)O NMR properties in various molecular models. The calculated (17)O NMR tensors are highly sensitive to the description of intermolecular hydrogen-bonding interactions at the target oxygen atom. A reasonably good agreement between experimental solid-state (17)O NMR data and B3LYP/6-311++G(d,p) calculations is achievable only in molecular cluster models where a complete hydrogen-bond network is considered. Using this theoretical approach, we also investigated the (17)O NMR tensors in two unusual structures: guanine- and uracil-quartets. PMID- 11853457 TI - Supramolecular gold nanoparticles for the redox recognition of oxoanions: syntheses, titrations, stereoelectronic effects, and selectivity. AB - Stable, CH(2)Cl(2)-soluble mixed dodecanethiol/(amidoferrocenyl)alkanethiol (AFAT) gold colloids were synthesized by ligand substitution reactions from Brust's dodecanethiol gold colloids and the AFAT ligands to study the recognition and titration of oxoanions. Gold colloids were obtained with various chain lengths (C(11) and C(6) chains) of the AFAT ligand and different proportions of AFAT ligands in the colloids. Modification of the amidoferrocenyl structure [replacement of the free C(5)H(5) ferrocene ring by C(5)Me(5) (Cp) or C(5)H(4)COCH(3)] has been achieved to investigate the stereoelectronic effects on the recognition. The cyclic voltammetry of these colloids in CH(2)Cl(2) on Pt electrode shows a reversible Fe(II/III) wave with some adsorption. With AFAT ligands, a new, less electrochemically reversible wave (with some adsorption) at a potential 220 +/- 20 mV less positive than that of the initial wave appears upon titration of [n-Bu(4)N][H(2)PO(4)], and the initial wave completely disappears after addition of 1 equiv of anion, which allows its titration. The potential shift does not depend on the AFAT proportion nor on its chain length but is reduced with Cp and enhanced with C(5)H(4)COCH(3), showing the key role of the hydrogen bonding between the -NH-amido group and a terminal oxygen atom of the oxoanions. According to the Echegoyen-Kaifer model, the potential shift leads to the ratio K((+))/K((0)) of apparent association constants. In the presence of both [n-Bu(4)N][HSO(4)] and [n-Bu(4)N]Cl, a shift of the initial wave (rather than its replacement) allows an easy titration, ideally with 20-Fc. Upon addition of [n-Bu(4)N][HSO(4)] alone, a weak wave shift (30 mV) of the colloids is also observed, allowing the titration of the HSO(4)(-) anion by the colloids containing a low percentage of AFAT ligand. The Echegoyen-Kaifer model provides access to the apparent association constant K((+)) in this case for which the interaction between the anion and the neutral form of the host is not significant. With the C(5)H(4)COCH(3) modification of the amidoferrocenyl branch, a new wave appears at a potential 70 mV more positive than the initial wave, signifying a stronger interaction with this modified ligand than with the parent AFAT ligand. These colloids favorably compare with ferrocenyl dendrimers in terms of rapid synthesis and selectivity of H(2)PO(4-) over HSO(4-) and with gold surfaces for the recognition of HSO(4)(-). PMID- 11853456 TI - Oxygen as a paramagnetic probe of membrane protein structure by cysteine mutagenesis and (19)F NMR spectroscopy. AB - Oxygen solubility increases toward the hydrophobic interior of membranes. Using NMR, this O(2) solubility gradient gives rise to an exquisite range of position dependent paramagnetic effects at partial pressures of 100 atm (PO(2)), which may be used to probe membrane protein structure and positioning. In this study, fluorinated probes were introduced at selected positions of the transmembrane 1 domain of the intact homotrimer of the integral membrane protein, diacylglycerol kinase. Using (19)F NMR, O(2)-induced chemical shift perturbations revealed secondary structure, membrane immersion depth, and regions of the helix in contact with the protein or with the micelle. PMID- 11853458 TI - A molecular electrostatic potential bond critical point model for atomic and group electronegativities. AB - A consistent set of atomic electronegativities of main block and d-block transition elements has been obtained from the position and value of the molecular electrostatic potential bond critical point of the C-E bond of a methyl element-hydride system, H(3)C-EH(n) (E is an element and n = 0, 1, 2, 3, 4, and 5 depending on the position of E in the periodic table). The new scale shows very good agreement with the popular electronegativity scales such as Pauling, Allen, Allred-Rochow, Mulliken, and Sanderson scales of electronegativity, especially for the main block elements. The present scale of electronegativity for transition elements is expected to be more accurate than the previously derived values because of a more consistent approach. Further, the same approach has led to the evaluation of group electronegativities when the hydrogens of E are replaced by other substituent groups. These group electronegativity values are found to correlate well with Inamoto and Mullay scales. PMID- 11853460 TI - Role of protein flexibility in enzymatic catalysis: quantum mechanical-molecular mechanical study of the deacylation reaction in class A beta-lactamases. AB - We present a theoretical study of a mechanism for the hydrolysis of the acyl enzyme complex formed by a class A beta-lactamase (TEM1) and an antibiotic (penicillanate), as a part of the process of antibiotic's inactivation by this type of enzymes. In the presented mechanism the carboxylate group of a particular residue (Glu166) activates a water molecule, accepting one of its protons, and afterward transfers this proton directly to the acylated serine residue (Ser70). In our study we employed a quantum mechanics (AM1)-molecular mechanics partition scheme (QM/MM) where all the atoms of the system were allowed to relax. For this purpose we used the GRACE procedure in which part of the system is used to define the Hessian matrix while the rest is relaxed at each step of the stationary structures search. By use of this computational scheme, the hydrolysis of the acyl-enzyme is described as a three-step process: The first step corresponds to the proton transfer from the hydrolytic water molecule to the carboxylate group of Glu166 and the subsequent formation of a tetrahedral adduct as a consequence of the attack of this activated water molecule to the carbonyl carbon atom of the beta-lactam. In the second step, the acyl-enzyme bond is broken, obtaining a negatively charged Ser70. In the last step this residue is protonated by means of a direct proton transfer from Glu166. The large mobility of Glu166, a residue that is placed in a Ohms-loop, is essential to facilitate this mechanism. The geometry of the acyl-enzyme complex shows a large distance between Glu166 and Ser70 and thus, if protein coordinates were kept frozen during the reaction path, it would be difficult to get a direct proton transfer between these two residues. This computational study shows how a flexible treatment suggests the feasibility of a mechanism that could have been discounted on the basis of crystallographic positions. PMID- 11853459 TI - Catalytic mechanism of heme oxygenase through EPR and ENDOR of cryoreduced oxy heme oxygenase and its Asp 140 mutants. AB - Heme oxygenase (HO) catalyzes the O(2)- and NADPH-cytochrome P450 reductase dependent conversion of heme to biliverdin, Fe, and CO through a process in which the heme participates both as a prosthetic group and as a substrate. In the present study, we have generated a detailed reaction cycle for the first monooxygenation step of HO catalysis, conversion of the heme to alpha-meso hydroxyheme. We employed EPR (using both (16)O(2) and (17)O(2)) and (1)H, (14)N ENDOR spectroscopies to characterize the intermediates generated by 77 K radiolytic cryoreduction and subsequent annealing of wild-type oxy-HO and D140A, F mutants. One-electron cryoreduction of oxy-HO yields a hydroperoxoferri-HO with g-tensor, g = [2.37, 2.187, 1.924]. Annealing of this species to 200 K is accompanied by spectroscopic changes that include the appearance of a new (1)H ENDOR signal, reflecting rearrangements in the active site. Kinetic measurements at 214 K reveal that the annealed hydroperoxoferri-HO species, denoted R, generates the ferri-alpha-meso-hydroxyheme product in a first-order reaction. Disruption of the H-bonding network within the distal pocket of HO by the alanine and phenylalanine mutations of residue D140 prevents product formation. The hydroperoxoferri-HO (D140A) instead undergoes heterolytic cleavage of the O-O bond, ultimately yielding an EPR-silent compound II-like species that does not form product. These results, which agree with earlier suggestions, establish that hydroperoxoferri-HO is indeed the reactive species, directly forming the alpha meso-hydroxyheme product by attack of the distal OH of the hydroperoxo moiety at the heme alpha-carbon. PMID- 11853461 TI - Molten gallium as a catalyst for the large-scale growth of highly aligned silica nanowires. AB - The vapor-liquid-solid (VLS) process is a fundamental mechanism for the growth of nanowires, in which a small size (5-100 nm in diameter), high melting point metal (such as gold and iron) catalyst particle directs the nanowire's growth direction and defines the diameter of the crystalline nanowire. In this article, we show that the large size (5-50 microm in diameter), low melting point gallium droplets can be used as an effective catalyst for the large-scale growth of highly aligned, closely packed silica nanowire bunches. Unlike any previously observed results using gold or iron as catalyst, the gallium-catalyzed VLS growth exhibits many amazing growth phenomena. The silica nanowires tend to grow batch by batch. For each batch, numerous nanowires simultaneously nucleate, grow at nearly the same rate and direction, and simultaneously stop growing. The force between the batches periodically lifts the gallium catalyst upward, forming two different kinds of products on a silicon wafer and alumina substrate. On the silicon wafer, carrot-shaped tubes whose walls are composed of highly aligned silica nanowires with diameters of 15-30 nm and length of 10-40 microm were obtained. On the alumina substrate, cometlike structures composed of highly oriented silica nanowires with diameters of 50-100 nm and length of 10-50 microm were formed. A growth model was proposed. The experimental results expand the VLS mechanism to a broader range. PMID- 11853462 TI - On the regioselectivity of the cyclization of enyne-ketenes: a computational investigation and comparison with the Myers-Saito and Schmittel reaction. AB - The Moore (C(2)-C(7)) cyclization and the alternative C(2)-C(6) cyclization of enyne-ketenes belong to the family of biradical cyclization reactions such as the Bergman reaction of ene-diynes, both the cyclizations of enyne-allenes and enyne cumulenes. The latter garnered substantial interest due to their antitumor efficacy. The mechanisms of both cyclization modes of enyne-ketenes are still unclear, but as the enyne-ketenes can formally be regarded as heteroanalogues of enyne-allenes, both cyclizations are expected to react via biradical routes. Nevertheless, as shown recently for cyclic allenes, the substitution of a methylene group by oxygen can lead to different energetic ordering of the electronic states of the key intermediates. To elucidate the mechanism, the present work investigates the course of both cyclization modes for various model compounds. To reveal general motifs for the large family of biradical cyclizations, a comparison with enyne-allenes is performed. PMID- 11853463 TI - Rapid determination of mercury in food colorants using electrothermal atomic absorption spectrometry with slurry sample introduction. AB - The determination of mercury in different types of color food additives using electrothermal atomic absorption spectrometry is proposed. The procedure involves direct introduction of the additive samples as suspensions, and fast-program methodology is used to avoid mercury volatilization losses. Suspensions are prepared in a medium containing 0.1% w/v Triton X-100, 1% v/v concentrated nitric acid, 2% w/v potassium permanganate, and 3% w/v silver nitrate before being directly introduced into the furnace. Calibration uses aqueous standards, and compensation of the background is carried out using the Zeeman device. The characteristic mass is 61 pg and the detection limit is 59 pg. The reliability of the procedure is checked by comparing the results obtained with other results based on microwave-oven sample digestion, and by analyzing a certified reference material. PMID- 11853465 TI - Statistical validation of the identification of tuna species: bootstrap analysis of mitochondrial DNA sequences. AB - Sequencing of the mitochondrial cytochrome b gene has been used to differentiate three tuna species: Thunnus albacares (yellowfin tuna), Thunnus obesus (bigeye tuna), and Katsuwonus pelamis (skipjack). A PCR amplified 528 bp fragment from 30 frozen samples and a 171 bp fragment from 26 canned samples of the three species were analyzed to determine the intraspecific variation and the positions with diagnostic value. Polymorphic sites between the species that did not present intraspecific variation were given a diagnostic value. The genetic distance between the sequences was calculated, and a phylogenetic tree was constructed, showing that the sequences belonging to the same species clustered together. The bootstrap test of confidence was used to determine the statistical validation of the species assignation, allowing for the first time a quantification of the certainty of the species assignation. The bootstrap values obtained from these results indicate that the sequencing of the cytochrome b fragments allows a correct species assignation with a probability > or =95%. PMID- 11853464 TI - Biosensor technology and surface plasmon resonance for real-time detection of genetically modified Roundup Ready soybean gene sequences. AB - Biospecific interaction analysis (BIA) was performed using surface plasmon resonance (SPR) and biosensor technologies to detect genetically modified Roundup Ready soybean gene sequences. We first immobilized, on SA sensor chips, single stranded biotinylated oligonucleotides containing soybean lectin and Roundup Ready gene sequences, and the efficiency of hybridization to oligonucleotide probes differing in length was determined. Second, we immobilized biotinylated PCR products from nontransgenic soybeans (genomes carrying only the lectin gene), as well as from genetically modified Roundup Ready soybean, and we injected the oligonucleotide probes. Furthermore, we used the sensor chips carrying either lectin and Roundup Ready soybean PCR products or 21-mer oligonucleotide as probes, and we injected both nonpurified and purified asymmetric PCR products. The results obtained show that 13 and 15 mer oligonucleotides are suitable probes to detect genetically modified Roundup Ready soybean gene sequences (either target oligonucleotides or PCR products) under standard BIA experimental conditions. By contrast, when 11 mer DNA probes were employed, no efficient hybridization was obtained. All the SPR-based formats were found to be useful for detection of Roundup Ready gene sequences, suggesting that these procedures are useful for the real-time monitoring of hybridization between target single stranded PCR products, obtained by using as substrates DNA isolated from normal or transgenic soybeans, and oligonucleotide or PCR-generated probes, therefore enabling a one-step, nonradioactive protocol to perform detection. PMID- 11853466 TI - (13)C NMR characterization of triacylglycerols of Moringa oleifera seed oil: an "oleic-vaccenic acid" oil. AB - The composition of acyl chains and their positions in the triacylglycerols of the oil extracted from seeds of Moringa oleifera were studied by (13)C NMR spectroscopy. The unsaturated chains of M. oleifera seed oil were found to comprise only mono-unsaturated fatty acids and, in particular, two omega-9 mono unsaturated acids, (cis-9-octadecenoic (oleic acid) and cis-11-eicosenoic acids) and one omega-7 mono-unsaturated acid (cis-11-octadecenoic acid (vaccenic acid)). The mono-unsaturated fatty acids were detected as separated resonances in the spectral regions where the carbonyl and olefinic carbons resonate according to the 1,3- and 2-positions on the glycerol backbone. The unambiguous detection of vaccenic acid was also achieved through the resonance of the omega-3 carbon. The (13)C NMR methodology enabled the simultaneous detection of oleate, vaccenate, and eicosenoate chains according to their positions on the glycerol backbone (1,3 and 2-positions) through the carboxyl, olefinic, and methylene envelope carbons of the triacylglycerol acyl chains. PMID- 11853467 TI - Differentiation of interglycosidic linkages in permethylated flavonoid glycosides from linked-scan mass spectra (B/E). AB - A series of per-O-methylated flavonoid di- and tri-glycosides, linked with 1-2 and/or 1-6 glycosidic bonds between sugar rings that were isolated from different plant materials were analyzed. It was demonstrated that the fragmentation behavior of permethylated flavonoid glycosides is dependent on the glycosidic bond placement between sugars. Y(n) type fragment ions, created after glycosidic bond cleavage with oxygen retention on sugar at the reducing end for permethylated compounds, were observed in the normal and linked-scan mass spectra recorded for alpha(1-2) bonded conjugates of flavonoid di- and tri-glycosides. Moreover, for alpha (1-6) linked glycosides, Y fragments created after rearrangement and elimination of internal sugar residues were observed in addition to Y(n) type ions, but these fragment ions were not registered in normal desorption ionization spectra. This second type of fragmentation was also reported previously in collision-induced dissociation tandem mass spectrometry (CID MS/MS) spectra of some oligosaccharides and flavonoid glycosides, but their presence was independent of the glycosidic bonds placement between sugar rings. PMID- 11853469 TI - Methylation methods for the quantitative analysis of conjugated linoleic acid (CLA) isomers in various lipid samples. AB - Precise methylation methods for various chemical forms of conjugated linoleic acid (CLA), which minimize the formation of t,t isomers and allylmethoxy derivatives (AMD) with the completion of methylation, were developed using a 50 mg lipid sample, 3 mL of 1.0 N H(2)SO(4)/methanol, and/or 3 mL of 20% tetramethylguanidine (TMG)/methanol solution(s). Free CLA (FCLA) was methylated with 1.0 N H(2)SO(4)/methanol (55 degrees C, 5 min). CLA esterified in safflower oil (CLA-SO) was methylated with 20% TMG/methanol (100 degrees C, 5 min), whereas CLA esterified in phospholipid (CLA-PL) was methylated with 20% TMG/methanol (100 degrees C, 10 min), followed by an additional reaction with 1.0 N H(2)SO(4)/methanol (55 degrees C, 5 min). Similarly, CLA esterified in egg yolk lipid (CLA-EYL) was methylated by base hydrolysis, followed by reaction with 1.0 N H(2)SO(4)/methanol (55 degrees C, 5 min). These results suggest that for the quantitative analysis of CLA in lipid samples by GC, proper methylation methods should be chosen on the basis of the chemical forms of CLA in samples. PMID- 11853468 TI - Analysis of intact glucosinolates by MALDI-TOF mass spectrometry. AB - Glucosinolates are naturally occurring plant compounds that may be important in the dietary prevention of cancer. This study shows that they can be detected in their intact form by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) with a high degree of sensitivity. The methodology was used to characterize a number of individual glucosinolates either produced by synthetic chemistry or isolated from plants. The method was used for crude plant extracts to rapidly examine the glucosinolate profile of the plant. The results for a range of plant extracts showed good agreement with previous LC MS analysis of the desulfoglucosinolates from the same samples. PMID- 11853470 TI - Improved amperometric method for the rapid and quantitative measurement of lipoxygenase activity in vegetable tissue crude homogenates. AB - An improved amperometric method for rapid (2 min) quantitative determination of lipoxygenase (LOX) activity in vegetable tissue crude homogenates is presented. Measured LOX activity was linear (R(2) > 0.99) throughout the entire activity range for green bean and for corn below 70% activity. The resolution was 0.4% or 1.11 micromol L(-1) s(-1) of oxygen. The limit of detection was 3.43 micromol L( 1) s(-1) of oxygen. The amperometric method was improved by encapsulating linoleic acid (LA) in beta-cyclodextrin (CD) resulting in a stable substrate buffer solution at a pH below 8.0. Ethanol and Tween 20 were not effective in solubilizing high LA concentrations required by the assay. A prototype benchtop instrument with the potential for use in an industrial environment is also presented. PMID- 11853471 TI - Determination of the pesticide napropamide in soil, pepper, and tomato by micelle stabilized room-temperature phosphorescence. AB - A selective and sensitive method for determining napropamide by room-temperature phosphorescence in SDS micelles is proposed and applied to the determination of this substance in a technical formulation and in spiked soil, pepper, and tomato samples. The use of phosphorescence enhancers such as sodium dodecyl sulfate (micellar agent), thallium (I) nitrate (external heavy atom), and sodium sulfite (deoxygenation agent) was studied and optimized to obtain maximum sensitivity. The determination was performed in 66 mM SDS, 30 mM thallium (I) nitrate, and 8 mM sodium sulfite. Taking into account both maximum phosphorescence intensity and the time required to reach that, a pH value of 7.2 was selected. After the samples were left standing at room temperature for 10 min, the phosphorescence was totally developed. The intensity was then measured at lambda(ex) = 282 nm and lambda(em) = 528 nm. The calibration graph was linear for 50-600 ng mL(-1) napropamide. The detection limit, according to the error propagation theory, was 16 ng mL(-1). The method has been demonstrated for the analysis of soils, peppers, and tomatoes, but, because of matrix interference, the method of standard additions was applied to determine napropamide in the vegetable samples. Recoveries from all these matrixes of added napropamide were near 100%. PMID- 11853473 TI - Detection of genetically modified maize by the polymerase chain reaction and capillary gel electrophoresis with UV detection and laser-induced fluorescence. AB - In this paper, the possibilities of capillary gel electrophoresis (CGE) to detect transgenic maize in flours are shown. The method is based on the extraction and amplification by the polymerase chain reaction (PCR) of a specific DNA fragment from transgenic maize and its subsequent analysis by CGE with UV detection or laser-induced fluorescence (LIF). Some useful considerations regarding the optimization of DNA extraction and amplification conditions are given. Also, a comparison is established between the two CGE protocols for DNA detection based on ultraviolet absorption (CGE-UV) and LIF (CGE-LIF). The requirements, advantages, and limitations of both CGE methods are discussed. To our knowledge, this is the first paper on the use of CGE-LIF to detect transgenic food. PMID- 11853472 TI - Hydrogen peroxide is required for poly(phenolic) domain formation during wound induced suberization. AB - The requirement for hydrogen peroxide (H(2)O(2)) during suberization was demonstrated in wound-induced potato tubers by monitoring the extent of phenolic polymerization after the inhibition of H(2)O(2) production using diphenyleneiodonium (DPI). In DPI-treated tissues the extent of phenolic polymerization in suberized tissues, measured using DFRC (Derivatization Followed by Reductive Cleavage) and thioglycolic acid analyses, was greatly reduced relative to untreated controls. Concomitantly, a large quantity of new soluble phenolics accumulated in the DPI-treated tissue some of which were not present in the controls. We suggest that the inhibition of H(2)O(2) production prevented these phenolics from being oxidized by cell wall peroxidases. As a result, these phenolics were left unpolymerized and accumulated in the tissue. Several of the soluble phenolics were identified as hydroxycinnamic acid derivatives. From the data presented, it was concluded that H(2)O(2) is required for the polymerization step in the formation of the poly(phenolic) domain of suberized potato tubers. PMID- 11853474 TI - Pyrolysis of lignin in the presence of tetramethylammonium hydroxide: a convenient method for S/G ratio determination. AB - Pyrolysis-gas chromatography in the presence of tetramethylammonium hydroxide (TMAH) was applied to the determination of the ratio of the abundances of the syringyl beta-aryl ether subunits to those of the guaiacyl equivalents (S/G) in lignin. Diazomethane-methylated kenafs (Hibiscus cannabinus and Hibiscus sabdariffa) and beech (Fagus crenata) in situ lignins were employed. Relative abundances of pyrolysis products derived from the guaiacyl and syringyl beta-aryl ether subunits were determined. The S/G ratios for in situ lignins were obtained with average 3.1% relative standard deviation for a minimum of six repeated runs. The S/G ratios determined by pyrolysis in the presence of TMAH agreed well with those determined by thioacidolysis, with a significant linear regression (R(2) = 0.9867). The results showed that pyrolysis with TMAH is an effective tool for obtaining information on the S/G ratio for in situ lignins. PMID- 11853476 TI - Method for the gas chromatographic assay with mass selective detection of trichloro compounds in corks and wines applied to elucidate the potential cause of cork taint. AB - To investigate the role of trichloro compounds as a potential cause of "cork taint" in wine, an assay for trichloroanisole (TCA) and trichlorophenol (TCP) in corks and wine was developed utilizing solid phase extraction on a C(18) cartridge followed by gas chromatography with mass selective detection. Recovery and imprecision for TCA were 86-102 and 1.6-5.8%, respectively, and for TCP 82 103% and 1.7-3.9%, respectively. Limits of detection and quantitation were 0.1 and 2 ng/L, respectively, for TCA, and 0.7 and 4 ng/L, respectively for TCP. A survey of 2400 commercial wines revealed a higher incidence of cork taint in white wine than in red and in wines utilizing composite cork closures; wines from central Europe and Spain had higher overall rates of contamination and those from Canada and Italy the lowest. Significant but modest associations were found between the TCA and TCP contents of the wines and corks, but many wines exhibiting cork taint had low or undetectable concentrations of TCA. Over a 12 month period, experimentally bottled wines exhibited a slow increase in TCA and TCP content while cork closures manifested a decrease; most bottles showing cork taint contained low levels of TCA, and TCP concentrations were well below the sensory threshold. Neither compound was cytotoxic to human cell lines in culture up to final concentrations of 500 ng/mL. It was concluded that these two trichloro compounds are, at most, minor components of cork taint in commercial wines. PMID- 11853475 TI - Authenticity assessment of estragole and methyl eugenol by on-line gas chromatography-isotope ratio mass spectrometry. AB - On-line capillary gas chromatography-isotope ratio mass spectrometry was used in the combustion (HRGC-C-IRMS) and the pyrolysis (HRGC-P-IRMS) modes to determine delta(13)C(PDB), delta(2)H(SMOW), and delta(18)O(SMOW) data of estragole (1) and methyl eugenol (2) originating from various sources. For 1, similar delta(13)C values, i.e., ranging from -35.4 to -29.9 per thousand and from -36.4 to -28.8 per thousand for the product of synthetic and natural origins, respectively, were found. The delta(2)H values ranged from -155 to -3 per thousand for synthetic 1 and from -193 to -105 per thousand for 1 from natural origin, whereas the determination of delta(18)O data gave values from +1.8 to +24.8 per thousand and from +2.7 to +18.7 per thousand for 1 from synthetic and natural origins, respectively. As synthetic 2 is produced by methylation of natural eugenol, the IRMS techniques did not allow differentiation of synthetic 2 from the product of natural origin. The recorded data ranges were nearly identical, i.e., delta(13)C = -37.4 to -35.0 per thousand and -41.1 to -32.2 per thousand; delta(2)H = -155 to -126 per thousand and -217 to -107 per thousand; delta(18)O = +5.5 to +6.6 per thousand and +2.7 to +6.9 per thousand, each for 2 from synthetic and natural origins, respectively. PMID- 11853477 TI - Microanalytical method for the characterization of fiber components and morphology of woody plants. AB - Microanalytical techniques were developed which allow the rapid characterization of fiber components and morphology of loblolly pine in a large number of samples. These techniques consist of extractives removal, holocellulose preparation, alpha cellulose and lignin content determination, and fiber length and coarseness analyses. Greater than 95% of the nonvolatile extractives from an increment core sample of loblolly pine was removed by four successive two-day acetone extractions. Fiber morphology and alpha-cellulose content was determined from holocellulose prepared from only 100 mg of wood. Similarly, a microanalytical acetyl bromide method was developed that enabled the accurate determination of lignin content from less than 50 mg of wood. Through the development of these microanlytical methods, it is possible to accurately and rapidly analyze fiber morphology and chemical components in a large number of increment core samples. PMID- 11853478 TI - Analysis and detection of the herbicides dimethenamid and flufenacet and their sulfonic and oxanilic acid degradates in natural water. AB - Dimethenamid [2-chloro-N-(2,4-dimethyl-3-thienyl)-N-(2-methoxy-1 methylethyl)acetamide] and flufenacet [N-(4-fluorophenyl)-N-(1-methylethyl)-2-(5 (trifluoromethyl)-1,3,4-thiadiazol-2-yl)oxy] were isolated by C-18 solid-phase extraction and separated from their ethanesulfonic acid (ESA) and oxanilic acid (OXA) degradates during their elution using ethyl acetate for the parent compound, followed by methanol for the polar degradates. The parent compounds were detected using gas chromatography-mass spectrometry in selected-ion mode. The ESA and OXA degradates were detected using high-performance liquid chromatography--electrospray mass spectrometry (HPLC-ESPMS) in negative-ion mode. The method detection limits for a 123-mL sample ranged from 0.01 to 0.07 microg/L. These methods are compatible with existing methods and thus allow for analysis of 17 commonly used herbicides and 18 of their degradation compounds with one extraction. In a study of herbicide transport near the mouth of the Mississippi River during 1999 and 2000, dimethenamid and its ESA and OXA degradates were detected in surface water samples during the annual spring flushes. For flufenacet, the only detections at the study site were for the ESA degradates in samples collected at the peak of the herbicide spring flush in 2000. The low frequency of detections in surface water likely is due to dimethenamid and flufenacet being relatively new herbicides. In addition, detectable amounts of the stable degradates have not been detected in ground water. PMID- 11853480 TI - Isolation and identification of organosulfur compounds oxidizing canine erythrocytes from garlic (Allium sativum). AB - Five compounds oxidizing canine erythrocytes were isolated from an aqueous ethanol garlic extract by silica gel column chromatography and preparative thin layer chromatography. On the basis of nuclear magnetic resonance, infrared spectroscopy, and mass spectrometry, they were identified as three known compounds: bis-2-propenyl trisulfide (1), bis-2-propenyl tetrasulfide (2), and bis-2-propenyl pentasulfide (3) as well as two novel compounds, bis-2-propenyl thiosulfonate (4) and trans-sulfuric acid allyl ester 3-allylsulfanyl-allyl ester (5). A mixture of compounds 1-3 and compounds 4 and 5 induced methemoglobin formation in canine erythrocyte suspension in vitro resulting in the oxidation of canine erythrocytes. These groups of characteristic organosulfur compounds contained in garlic probably contribute to oxidations in blood. The constituents of garlic have the potential to oxidize erythrocytes and hemoglobin, suggesting that foods containing quantities of garlic should be avoided for feeding dogs. PMID- 11853479 TI - Natural zinniol derivatives from Alternaria tagetica. Isolation, synthesis, and structure-activity correlation. AB - Two novel phytotoxins, 8-zinniol methyl ether (5) and 8-zinniol acetate (6), in addition to 6-(3',3'-dimethylallyloxy)-4-methoxy-5-methylphthalide (2), 5-(3',3' dimethylallyloxy)-7-methoxy-6-methylphthalide (3), and the novel metabolites 8 zinniol 2-(phenyl)ethyl ether (4) and 7-zinniol acetate (7) have been identified as natural zinniol derivatives from the organic crude extract of Alternaria tagetica culture filtrates. Using zinniol as the starting material, phytotoxin 5 was synthesized, together with a number of synthetic intermediates (8-13). Both natural and synthetic zinniol derivatives were evaluated in the leaf-spot bioassay against marigold leaves (Tagetes erecta). PMID- 11853482 TI - Isolation and purification of lutein from the microalga Chlorella vulgaris by extraction after saponification. AB - A simple and efficient method for the isolation and purification of lutein from the microalga Chlorella vulgaris was developed. Crude lutein was obtained by extraction with dichloromethane from the microalga after saponification. Partition values of lutein in the two-phase system of ethanol-water dichloromethane at different ratios were measured by HPLC so as to assist the determination of an appropriate condition for washing water-soluble impurities in the crude lutein. Partition values of lutein in another two-phase system of ethanol-water-hexane at different ratios were also measured by HPLC for determining the condition for removing fat-soluble impurities. The water-soluble impurities in the crude lutein were removed by washing with 30% aqueous ethanol, and the fat-soluble impurities were removed by extraction with hexane. The final purity of lutein obtained was 90-98%, and the yield was 85-91%. PMID- 11853483 TI - Conversion of dextran to cycloisomaltooligosaccharides using an enzyme immobilized porous hollow-fiber membrane. AB - This paper describes a cycloisomaltooligosaccharide glucanotransferase (CITase) multilayer-immobilized porous hollow-fiber membrane used as an enzyme bioreactor. Dextran, a substrate with an average molecular mass of 43000, is converted into seven- to nine-glucose-membered cycloisomaltooligosaccharides, effective as a preventive for dental caries, aided by convective transport of the substrate to the vicinity of the enzyme through the pores. Epoxy-group-containing graft chains were uniformly appended onto the surface of pores throughout a porous hollow fiber membrane by radiation-induced graft polymerization. Subsequently, a diethylamino group was introduced, as an anion-exchange moiety, to the graft chains, which caused the chains to expand toward the interior of the pores due to mutual electrostatic repulsion. The expanding graft chain provided multilayer binding sites for CITase. Fifty-five milligrams of adsorbed CITase per gram of membrane is equivalent to the degree of multilayer binding of 5. Finally, 80% of the multilayer-adsorbed CITase was immobilized via enzymatic cross-linking with transglutaminase to prevent the leakage of enzymes. CITase, with a degree of multilayer immobilization of 4, produced the target cycloisomaltooligosaccharides at a conversion yield of 55% in weight at 310 K during permeation by the dextran solution at a space velocity defined as the permeation rate divided by membrane volume of 6 per hour. PMID- 11853481 TI - Composition of a chemopreventive proanthocyanidin-rich fraction from cranberry fruits responsible for the inhibition of 12-O-tetradecanoyl phorbol-13-acetate (TPA)-induced ornithine decarboxylase (ODC) activity. AB - Phenolics from the American cranberry (Vaccinium macrocarpon) were fractionated into a series of proanthocyanidins and other flavonoid compounds by vacuum chromatography on a hydrophilic, porous polyvinylic gel permeation polymer. Antioxidant activity was not restricted to a particular class of components in the extract but was found in a wide range of the fractions. Significant chemopreventive activity, as indicated by an ornithine decarboxylase assay, was localized in one particular proanthocyanidin-rich fraction from the initial fractionation procedure. Further fractionation of the active anticarcinogenic fraction revealed the following components: seven flavonoids, mainly quercetin, myricetin, the corresponding 3-O-glycosides, (-)-epicatechin, (+)-catechin, and dimers of both gallocatechin and epigallocatechin types, and a series of oligomeric proanthocyanidins. PMID- 11853484 TI - Changes occurring in phenolic compounds and alpha-tocopherol of virgin olive oil during storage. AB - Changes occurring in the concentrations of alpha-tocopherol, total phenols, and complex phenols linked to 3,4-dihydroxyphenylethanol (fractions FII and FIV) and p-hydroxyphenylethanol (FIII) during storage of virgin olive oil under environmental conditions were studied. Under diffused light, alpha-tocopherol was decomposed by 79% in 4 months, whereas <45% of the phenols were lost during the same period. Among the phenols, FII showed the least stability, and decreased by 72% in 6 months. Total phenols, FIII, and FIV recorded reductions in the range of 57-63% in 6 months. When the oil was stored in the dark, alpha-tocopherol, total phenols, FIII, and FIV exhibited similar profiles of degradation, reducing by 39 45% in the first 6 months and 50-62% in 12 months. FII was the least stable compound in the dark and recorded a loss of 64% in 6 months and 79% in 12 months. The levels of the above antioxidants were further related to peroxide formation. Remaining levels of these compounds at PV = 20 meq/kg ranged between 50 and 73% under diffused light and between 40 and 62% in the dark. PMID- 11853486 TI - Disappearance of pyrimethanil residues on tomato plants. AB - The objective of this work was to estimate the disappearance of pyrimethanil, the active ingredient of Mythos 300 SC, at present, commonly used for the protection of greenhouse vegetables against diseases of fungal origin. Disappearance trends of the chemical deposits were studied on tomato plants grown in commercial greenhouses sprayed with a homogeneous 0.15% aqueous solution of the plant protection product. It was found that, on average, pyrimethanil residues on ripening fruits dropped by half and reached detection limit level in 5.7 and 13.7 days after Mythos 300 SC application, respectively. Pyrimethanil residues on tomato leaves dropped by half within 4.0 days and reached detection limit level in the first 10.5 days. Therefore, in conditions of high infection pressure, there is a need to repeat the fungicide application as early as after 3 or 4 days after previous application of Mythos 300 SC. PMID- 11853485 TI - Simulated sunlight-induced photodegradations of triasulfuron and cinosulfuron in aqueous solutions. AB - To elucidate the photochemical behavior of two sulfonylureas (cinosulfuron and triasulfuron) for which the chemical formulas are relatively close, their photodegradation was studied in water. All experiments were carried out under laboratory conditions using a xenon arc lamp as the source of radiation to simulate environmental conditions. Polychromatic quantum efficiencies were calculated to determine the photochemical pesticide lifetimes at pH 7, and a comparison with hydrolysis lifetimes has been performed. The results obtained showed clearly that at pH 7, photodegradation becomes a more important pathway than chemical degradation. HPLC-DAD was used to study the kinetics for both sulfonylureas and their photoproducts, whereas HPLC-MS (ESI in positive and negative modes) was used to identify photoproducts. These results suggest that the photodegradation of these two sulfonylureas proceeds via a number of reaction pathways: (1) cleavage of the sulfonylurea bridge; (2) desulfonylation, which can proceed either by a carbon-sulfur cleavage or a nitrogen-sulfur cleavage; (3) O demethylation of methoxy moieties present on the triazine ring; and (4) O dealkylation of benzene derivatives. In addition, it was found that the desulfonylation represented the main step and that it was wavelength dependent. PMID- 11853487 TI - Spontaneous formation of thiuram disulfides in solutions of iron(III) dithiocarbamates. AB - Dithiocarbamates are used as pesticides and rubber additives. Dithiocarbamates are the reduced forms of thiuram disulfides and both of these groups of substances induce allergic contact dermatitis. The allergic cross-reactivity pattern between dithiocarbamates and thiurams is unclear. The aim of this study was to investigate why these cross-reactions occur sometimes but not always. HPLC analysis of buffer solutions of iron(III) dithiocarbamates demonstrated that thiuram disulfides were formed spontaneously and rapidly in high yield. No such oxidation was observed in solutions of copper(II), zinc(II), or sodium dithiocarbamates. However, sodium diethyldithiocarbamate and zinc diethyldithiocarbamate were oxidized in buffer solution when ferric salt was added. The influence of different metal ions on the oxidation reaction is probably an explanation for the cross-reactivity patterns seen between dithiocarbamates and thiurams. These findings also show that careful handling is necessary in analytical and biological studies with solutions of iron(III) dithiocarbamates. Oxidation of dithiocarbamates in aqueous buffer at physiological pH has not been shown before. PMID- 11853488 TI - Characterization of cider apples on the basis of their fatty acid profiles. AB - In the current study, the fatty acids composition of 30 monovarietal apple juices from six cider apple varieties belonging to two categories was analyzed. The different apple juices were obtained from three consecutive harvests (1997, 1998, and 1999). The fatty acids concentration in apple juice together with chemometric techniques such as principal components analysis (PCA), soft independent modeling of class analogy (SIMCA), and linear discriminant analysis (LDA), allowed us to differentiate apple juices on the basis of the sweet or sharp category to which the cider apple variety belongs. Fatty acids such as the unsaturated oleic and linoleic acids, and saturated caprylic, capric, stearic, and palmitic acids were related to the sweet cider apple category, while pentadecanoic acid is related to the sharp class. PMID- 11853489 TI - Comparison of key aroma compounds in cooked brown rice varieties based on aroma extract dilution analyses. AB - The aroma compounds present in cooked brown rice of the three varieties Improved Malagkit Sungsong (IMS), Basmati 370 (B 370), and Khaskhani (KK), and of the variety Indica (German supermarket sample), were identified on the basis of aroma extract dilution analyses (AEDA). A total of 41 odor-active compounds were identified, of which eleven are reported for the first time as rice constituents. 2-Amino acetophenone (medicinal, phenolic), which was up to now unknown in rice aroma, exhibited the highest flavor dilution (FD) factor among the 30 to 39 odor active compounds detected in all four varieties. 2-Acetyl-1-pyrroline, exhibiting an intense popcorn-like aroma-note, was confirmed as a further key aroma constituent in IMS, B 370, and KK, but was not important in Indica. Differences in the FD factors between the varieties were found for the previously unknown rice aroma compound 3-hydroxy-4,5-dimethyl-2(5H)-furanone (Sotolon; seasoning like), which was higher in B 370 than in IMS and KK. In IMS, a yet unknown, spicy smelling component with a very high FD factor could be detected, which contributed with lower FD factors to the overall aromas of B 370 and KK, and was not present in Indica. The latter variety, which was available on the German market, differed most in its overall aroma from the three Asian brown rices. PMID- 11853490 TI - Transfer of aroma compounds through the lipidic-aqueous interface in a complex system. AB - The transfer kinetics of aroma compounds from the aqueous phase to the lipidic phase (miglyol) and from miglyol to the aqueous phase have been studied in the presence or absence of a protein, beta-lactoglobulin, and at different pH values. In the presence of beta-lactoglobulin, the transfer at the interface from the aqueous to the lipidic phase decreases, with a greater effect of the presence of the protein at pH 3 than at pH 6. This barrier effect of the protein plays a role in the transfer of the aroma compounds between the different phases of the matrix. PMID- 11853491 TI - Retronasal transport of aroma compounds. AB - A comparison was made between the amounts of volatiles in the headspace above a solution and the breath volatile content (exhaled from the nose or mouth) after consumption of the same solution. The amounts of volatiles in the breath were lower than those in the headspace, with breath exhaled via the mouth containing, on average, 8-fold more volatiles than breath exhaled via the nose. Dilution of the sample by saliva in-mouth did not appear to be a major factor affecting volatile delivery. Instead, the rate of in vivo equilibration (mass transfer) appeared to be the most significant factor, principally affecting volatile delivery from the solution to the gas phase. Thereafter, gas-phase dilution of the volatile as it passed through the upper airway resulted in a further decrease in volatile concentration. The final factor affecting the volatile concentration exhaled from the nose was absorption of volatiles to the nasal epithelia, which was greatest for those compounds with the lowest air/water partition coefficients. PMID- 11853492 TI - Effect of temperature and pH on the generation of flavor volatiles in extrusion cooking of wheat flour. AB - Extrusion temperature (120, 135, and 150 degrees C) and quantity of added sodium hydroxide (0, 3, and 6 g/kg feedstock) were used as variables to study flavor generation in extrusion cooking of wheat flour. In total, 127 volatile components were identified in the extrudates, of which 51 contained sulfur. The levels of pyrroles, thiophenes, thiophenones, thiapyrans, and thiazolines increased at higher extrusion temperatures, whereas furans and aldehydes decreased. The addition of sodium hydroxide also affected the formation of volatile compounds. However, thiophenes, thiophenones, polythiacycloalkanes, thiazoles, thiazolines, pyrroles, and some pyrazines tended to increase with the more alkaline extrusion conditions. Some compounds from lipid-Maillard interactions were identified in the extrudates. Analysis of the volatile components by gas chromatography olfactometry showed sulfur- and nitrogen-sulfur-containing heterocycles as possible contributors to the sulfury and rubbery odors observed in extrudates produced at the higher temperature and more alkaline conditions. PMID- 11853493 TI - Effect of lipid composition on meat-like model systems containing cysteine, ribose, and polyunsaturated fatty acids. AB - This paper compares the volatile constituents of model systems containing the important meat aroma precursors cysteine and ribose, with and without either methyl linoleate, an n-6 fatty acid, or methyl alpha-linolenate, an n-3 acid, both of which are present in meat. Many of the volatile compounds formed from the reaction between cysteine and ribose were not formed, or formed in lower amounts, when lipid was present. This may be due to the reaction between hydrogen sulfide, formed from the breakdown of cysteine, and lipid degradation products. In addition, cysteine and ribose modified lipid oxidation pathways, so that alcohols and alkylfurans were formed rather than saturated and unsaturated aldehydes. Several volatile compounds, which have been found at elevated levels in cooked meat from animals fed supplements high in n-3 acids, were formed when methyl alpha-linolenate reacted with cysteine and ribose. The possible effects of increasing the n-3 content of meat upon flavor formation during cooking are discussed. PMID- 11853494 TI - Volatile compounds of selected strawberry varieties analyzed by purge-and-trap headspace GC-MS. AB - Six strawberry (Fragaria x ananassa Duch.) varieties, Senga Sengana, Jonsok, Polka, Korona, Bounty, and Honeoye, were studied in relation to composition of their volatile headspace compounds collected by dynamic purge-and-trap concentration. Also, the influence of cultivation technique on the volatile compounds was tested by comparing conventionally and organically cultivated strawberry varieties. In addition, the effect of geographical origin on strawberry volatiles was investigated. The emphasis of this study was lay in the industrial utilization of strawberry. A total of 52 different volatile compounds were determined from frozen strawberries grown in 1997 and 1998. In addition to individual compounds, total areas of chromatogram profiles were compared. The major esters were found to be methyl butanoate, ethyl butanoate, methyl hexanoate, and ethyl hexanoate. With the application of principal component analysis and analysis of variance to the data matrix, the varieties were classified into three groups. Var. Honeoye was most different from the others. Organic cultivation had no effect on strawberry volatiles. PMID- 11853495 TI - Chemometric analysis of Ragusano cheese flavor. AB - Ragusano cheeses were produced in duplicate from milk collected from pasture-fed and total mixed ration (TMR)-fed cattle at four time intervals. The cheeses were subjected to chemical analysis, conventional sensory testing, and gas chromatography-olfactometry (GCO). Data from each type of analysis were examined by principal component and factor analysis and by pattern recognition (SIMCA) to see if sufficient information for classification into pasture-fed and TMR-fed groups was contained therein. The results clearly indicate that there are significant differences in sensory panel and chemical analysis results between the two cheeses. The data were also examined to see if models of sensory responses as a function of analytical or GCO results or both could be constructed with the modeling technique partial least-squares regression (PLS). Strong PLS models of some sensory responses (green and toasted odor; salt, pungent, bitter, and butyric sensations; and smooth consistency) were obtained. PMID- 11853496 TI - Antithrombotic lipid minor constituents from vegetable oils. Comparison between olive oils and others. AB - Many epidemiological studies suggest that vegetable oils and especially olive oil present a protective effect against atherosclerosis. In this study, total lipids (TL) of Greek olive oils and seed oils of four kinds, namely, soybean, corn, sunflower, and sesame oil, were separated into total polar lipids (TPL) and total neutral lipids (TNL) via a novel extraction procedure. TPL and TNL of olive oil were fractionated by HPLC for further study. Each lipid fraction from HPLC separation along with TL, TPL, and TNL lipid samples from oils were tested in vitro for their capacity to induce or to inhibit washed rabbit platelet aggregation. Comparison between olive and seed oils supports the superiority of olive oil as high levels of platelet activating factor (PAF) antagonists have been detected, mainly in TPL. In addition, the structure of the most active fraction from olive oil was elucidated, as a glycerol-glycolipid. Because it has already been reported that PAF plays a pivotal role in atherogenesis, the existence of PAF agonists and antagonists in vegetable oils may explain their protective role against atherosclerosis. PMID- 11853497 TI - Toxic potency of polychlorinated dibenzo-p-dioxins, polychlorinated dibenzofurans, and polychlorinated biphenyls in food samples from Catalonia (Spain). AB - A surveillance program on polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), and polychlorinated biphenyls (PCBs) in 29 foodstuff samples produced all over the four provinces in Catalonia (Spain) is presented. The study included the analyses of milk, egg, meat (beef, chicken, and pork), mussel, and olive oil samples. A previously developed method for the simultaneous analysis of the 2,3,7,8-substituted PCDDs/PCDFs and the dioxin-like PCBs, as well as the indicator PCBs, was employed. Total toxicity equivalent (TEQ) values were calculated using the toxicity equivalent factors (TEFs) proposed by the World Health Organization (WHO) for dioxin-like PCBs, PCDDs, and PCDFs. The TEQ(PCDD/F) levels were below the limits proposed in the draft of the EC regulation for food commercialization in the European countries. These limits are the following: 2 pg WHO-TEQ/g fat for pork, 3 pg WHO-TEQ/g fat for milk and chicken, 5 pg WHO-TEQ/g fat for egg and beef, and 3 pg WHO-TEQ/g whole product for fish. The contributions of PCDDs/Fs and dioxin-like PCBs in the total toxicity of the samples were calculated for each matrix. The results showed that the TEQ(PCB) contribution varied from 27% in olive oil samples to 81% in mussel samples. These findings suggest that the regulation of TEQ contents in food should include not only the TEQ(PCDD/F), but also the TEQ(PCB). PMID- 11853498 TI - Evaluation of extracts and isolated fraction from Capparis spinosa L. buds as an antioxidant source. AB - The antioxidant activity of extracts from Capparis spinosa L. buds was evaluated using different in vitro tests: ascorbate/Fe(2+)-mediated lipid peroxidation of microsomes from rat liver; bleaching of 1,1-diphenyl-2-picryl-hydrazyl radical; and autoxidation of Fe(2+) ion in the presence of bathophenanthroline disulfonate. The methanolic extract showed strong activities in all of these in vitro tests. The amount of total phenols was determined in the methanolic extract. In addition, the level of rutin was calculated as 0.39% (w/w) by HPLC analysis. Our findings indicate the following: (a) the antioxidant efficiency of the methanolic extract may be attributed to its phenolic content; and (b) the antioxidant activity of the methanolic extract was maintained after removal of glucosinolates, confirming that these compounds do not interfere with the antioxidant properties of the extract. The results obtained from this study exalt the nutritional value of the flowering buds (capers) which are widely used as a source of flavor. PMID- 11853499 TI - Residues of methamidofos, malathion, and methiocarb in greenhouse crops. AB - The diminution of methamidofos, malathion, and methiocarb in different crops grown in greenhouses has been studied, including the presence of metabolites such as malaoxon, methiocarb sulfoxide, and methiocarb sulfone. The analytical method is based on dichloromethane extraction and GC-PFPD analysis. It has been validated establishing performance parameters such as recovery rates, precision, linear ranges, and limits of detection and quantification, which are low enough for ensuring that their corresponding MLRs can be adequately quantified. Samples of treated cucumbers and peppers grown in greenhouses were collected and analyzed during a 15-day period for obtaining the diminution rates of methamidofos and malathion. The behavior of methiocarb in treated green beans and tomatoes has been studied using analysis of variance (ANOVA) as the statistical tool, for establishing the influence of crop, season, application dose, and greenhouse design. PMID- 11853500 TI - Quantification of melanoidin concentration in sugar-casein systems. AB - Melanoidins are the final, brown, high molecular weight products of the Maillard reaction. The aim of the present study was to determine the average molar extinction coefficients of melanoidins formed in heated glucose-casein and fructose-casein systems. The value of the extinction coefficient can be used to translate spectrophotometrically measured browning (absorbance values) into melanoidin concentration. In the present study the melanoidins were quantified by measuring the concentration of sugar incorporated into the melanoidins, using (14)C-labeled sugar. The extinction coefficient of the melanoidins remained constant during the observation period as the absorbance at 420 nm increased to approximately 8 units, and it was calculated to be 477 (+/- 50) L mol(-1) cm(-1) in the glucose-casein reaction and 527 (+/- 35) L mol(-1) cm(-1) in the fructose casein reaction. This difference is not significant. An increase of the number of sugar molecules per reactive amino group during the heating of glucose-casein and the fructose-casein mixtures was observed by the radiochemical method as well as by microanalysis of the high molecular weight fraction. PMID- 11853501 TI - Interactions between flavonoids and proteins: effect on the total antioxidant capacity. AB - Flavonoids are potent antioxidants. It is also known that flavonoids bind to proteins. The effect of the interaction between tea flavonoids and proteins on the antioxidant capacity was examined. Their separate and combined antioxidant capacities were measured with the Trolox equivalent antioxidant capacity (TEAC) assay. It was observed that the antioxidant capacity of several components of green and black tea with alpha-, beta-, and kappa-casein or albumin is not additive; that is, a part of the total antioxidant capacity is masked by the interaction. This masking depends on both the protein and the flavonoid used. Components in green and black tea, which show the highest masking in combination with beta-casein, are epigallocatechin gallate and gallic acid. The results demonstrate that the matrix influences the efficacy of an antioxidant. PMID- 11853502 TI - Capsaicinoids in vegetative organs of Capsicum annuum L. in relation to fruiting. AB - Pepper (cv. Padron) shows a spatial gradient in the content of phenolic compounds, and particularly of capsaicinoids, along the stem. These compounds were consistently more abundant in apical fruits than in fruits belonging to middle and basal segments. Analysis of the two principal capsaicinoids in fruits showed that the proportion of capsaicin was always higher than that of dihydrocapsaicin. Capsaicinoids were also found to be present in vegetative organs, such as stem and leaves. In this case, the proportion of individual capsaicinoids was different than that in fruits, and dihydrocapsaicin was found to be more abundant. To find out whether the capsaicinoids in vegetative organs came from the fruits, the floral buds were removed and fruit formation was prevented. Capsaicinoids were not detected in the stem and leaves of floral bud deprived plants, suggesting that they did originate from the fruit. PMID- 11853503 TI - Alkylpyridiniums. 1. Formation in model systems via thermal degradation of trigonelline. AB - Trigonelline is a well-known precursor of flavor/aroma compounds in coffee and undergoes significant degradation during roasting. This study investigates the major nonvolatile products that are procured after trigonelline has been subjected to mild pyrolysis conditions (220-250 degrees C) under atmospheric pressure. Various salt forms of trigonelline were also prepared and the thermally produced nonvolatiles analyzed by thin layer chromatography, liquid chromatography-electrospray ionization tandem mass spectrometry, and (1)H and (13)C nuclear magnetic resonance. Results revealed the decarboxylated derivative 1-methylpyridinium as a major product of certain salts, the formation of which is positively correlated to temperature from 220 to 245 degrees C. Moreover, trigonelline hydrochloride afforded far greater amounts of 1-methylpyridinium compared to the monohydrate over the temperature range studied. Investigations into other potential quaternary amine products of trigonelline also indicate nucleophilic substitution reactions that lead to dialkylpyridiniums, albeit at concentration levels approximately 100-fold lower than those recorded for 1 methylpyridinium. PMID- 11853505 TI - Stabilization of oil-in-water emulsions by beta-lactoglobulin-polyethylene glycol conjugates. AB - The disulfide bonds of beta-lactoglobulin (beta-lg) were modified by oxidative sulfitolysis to generate beta-lgSO(3). The native protein (beta-lg) and the modified protein (beta-lgSO(3)) were conjugated to activated polyethylene glycol (PEG) to generate beta-lgPEG and beta-lgSO(3)PEG, respectively. Oil-in-water (o/w) emulsions containing 1% beta-lg or beta-lg conjugates were prepared at pH 2.8, 5.0, and 7.0. Emulsion droplet diameters and zeta potentials were measured. For the same emulsifier, emulsion droplet diameters decreased when emulsion pH increased. Zeta potentials of emulsion droplets increased with pH for beta-lg and beta-lgSO(3). Zeta potentials of beta-lgPEG and beta-lgSO(3)PEG approached zero, suggesting that the protein molecule was covered by PEG chains. Accelerated and 7 day storage stabilities at 21 degrees C of the emulsions were monitored. The emulsifying activity index (EAI) of beta-lgPEG was not significantly different from the EAI of beta-lg. The EAI of beta-lg was enhanced following sulfitolysis of beta-lactoglobulin. The emulsifying activity increased more when the oxidatively modified protein was conjugated to polyethylene glycol. Emulsions made with beta-lgSO(3)PEG were more stable than emulsions made with beta-lg, beta lgPEG, or beta-lgSO(3) under accelerated stability study and for 7 days at 21 degrees C. The stability of o/w emulsions stabilized with beta-lgSO(3)PEG increased because individual droplets were better protected, against protein bridging or coalescence, by the thick adsorbed protein-PEG layer. PMID- 11853504 TI - Alkylpyridiniums. 2. Isolation and quantification in roasted and ground coffees. AB - Recent model studies on trigonelline decomposition have identified nonvolatile alkylpyridiniums as major reaction products under certain physicochemical conditions. The quaternary base 1-methylpyridinium was isolated from roasted and ground coffee and purified by ion exchange and thin-layer chromatography. The compound was characterized by nuclear magnetic resonance spectroscopy ((1)H, (13)C) and mass spectrometry techniques. A liquid chromatography-electrospray ionization tandem mass spectrometry method was developed to quantify the alkaloid in coffee by isotope dilution mass spectrometry. The formation of alkylpyridiniums is positively correlated to the roasting degree in arabica coffee, and highest levels of 1-methylpyridinium, reaching up to 0.25% on a per weight basis, were found in dark roasted coffee beans. Analyses of coffee extracts also showed the presence of dimethylpyridinium, at concentrations ranging from 5 to 25 mg/kg. This is the first report on the isolation and quantification of alkylpyridiniums in coffee. These compounds, described here in detail for the first time, may have an impact on the flavor/aroma profile of coffee directly (e.g., bitterness), or indirectly as precursors, and potentially open new avenues in the flavor/aroma modulation of coffee. PMID- 11853507 TI - Comparative study of the products of the peroxidase-catalyzed and the polyphenoloxidase-catalyzed (+)-catechin oxidation. Their possible implications in strawberry (Fragaria x ananassa) browning reactions. AB - The peroxidase- and polyphenoloxidase-catalyzed oxidations of (+)-catechin yield several products showing different degrees of polymerization, which are apparently responsible for the pigment decay and the associated browning reaction that occurs in processed strawberry fruits and their derived foods. In this work, we have purified both peroxidase and polyphenoloxidase from Oso Grande cv. strawberry fruits, and comparatively analyzed the products of their enzyme mediated (+)-catechin oxidation. The joint analysis by reversed-phase and size exclusion HPLC of the (+)-catechin oxidation products obtained with both enzymes indicate that they were qualitatively the same: dehydrodicatechin B4, a (+) catechin quinone methide, dehydrodicatechin A, a (+)-catechin trimer, and a (+) catechin oligomer with polymerization degree equal to or greater than 5. The main quantitative differences between the oxidative reactions were the great amount of oligomer formed in the case of the polyphenoloxidase-mediated reaction and the low amount of (+)-catechin reacted in the case of the peroxidase-mediated reaction. One of the possible reasons for such low levels of (+)-catechin consumption in the case of the peroxidase-mediated reaction was the possible inhibition by products of the enzyme-catalyzed oxidation. In fact, the peroxidase mediated (+)-catechin oxidation was differentially inhibited by dehydrodicatechin A, showing a competitive type inhibition and a k(I) of 6.4 microM. In light of these observations, these results suggest that brown polymer formation, estimated as oligomeric compounds resulting from (+)-catechin oxidation, in strawberries is mainly due to polyphenoloxidase, and although peroxidase also plays an important role, it is apparently auto-regulated by product (dehydrodicatechin A) inhibition. PMID- 11853506 TI - Antioxidant properties of trans-epsilon-viniferin as compared to stilbene derivatives in aqueous and nonaqueous media. AB - trans-Epsilon-viniferin, the dimer of resveratrol, extracted from Vitis vinifera, has been evaluated for its antioxidant capacity. Its properties have been compared to those of resveratrol and synthetic stilbenic derivatives (4 hydroxystilbene, 4,4'-dihydroxystilbene, 3,5-dihydroxystilbene, and trimethylresveratrol), in regard to their liposolubility using two media with different polarity. The bleaching of beta-carotene by lipoperoxyl (LOO.) radicals in an oil/water (O/W) emulsion and the scavenging of superoxide anions (O(-)(2) in dimethyl sulfoxide (DMSO) using 5,5-dimethyl-1-pyrroline-N-oxide as a spin trap were followed using UV-visible and electron paramagnetic resonance, respectively. Epsilon-viniferin exhibits the best antioxidant capacity in the DMSO/O(-)(2) polar system (IC(50) = 0.14 mM) while 4,4'-dihydroxystilbene presents the highest antioxidant capacity in the O/W/LOO. system (inhibition of beta-carotene bleaching, 82%). Partition coefficients and kinetics of partition between 1-octanol and water were measured to discuss the antioxidant efficiency of the compounds in relation with their chemical structure. PMID- 11853508 TI - Antiadhesive effect of green and roasted coffee on Streptococcus mutans' adhesive properties on saliva-coated hydroxyapatite beads. AB - Green and roasted coffees of the two most used species, Coffea arabica and Coffea robusta, several commercial coffee samples, and known coffee components were analyzed for their ability to interfere with Streptococcus mutans' sucrose independent adsorption to saliva-coated hydroxyapatite (HA) beads. All coffee solutions showed high antiadhesive properties. The inhibition of S. mutans' adsorption to HA beads was observed both when coffee was present in the adsorption mixture and when it was used to pretreat the beads, suggesting that coffee active molecules may adsorb to a host surface, preventing the tooth receptor from interacting with any bacterial adhesions. Among the known tested coffee components, trigonelline and nicotinic and chlorogenic acids have been shown to be very active. Dialysis separation of roasted coffee components also showed that a coffee component fraction with 1000 Da < MW < 3500 Da, commonly considered as low MW coffee melanoidins, may sensibly contribute to the roasted coffee's antiadhesive properties. The obtained results showed that all coffee solutions have antiadhesive properties, which are due to both naturally occurring and roasting-induced molecules. PMID- 11853510 TI - Coagulation of soy protein isolates induced by subtilisin Carlsberg. AB - The coagulation of soy protein isolates (SPI) induced by subtilisin Carlsberg was studied. The proteins were digested to fragments of 16 kDa or less in the early stage of the reaction, followed by coagulation. The time-course of the coagulation measured by turbidity was separated into three phases. The turbidity decreased from the initial level observed at time zero to the minimum level (OD1) at time T1 (15-20 min) in the first phase. Then, it increased drastically to reach the maximum (OD2) at time T2 (60-70 min) in the second phase, which was followed by a slight decrease in the third phase. The coagulation was terminated at T2, where 30-35% of the weight of the SPI proteins was in coagula. Proteins in the coagula were degraded slowly in the prolonged incubation, and the protein content in the coagula was finally 15-20% of the weight. The time-course of the turbidity agreed well with that of the weight of the precipitates formed, indicating that the turbidity reflects the progress of the coagulation. The turbidity change (OD1 to OD2) from the start to the end of the coagulation increased proportionally to the SPI concentration (4.9-11 mg/mL), although the time (T1 to T2) needed for the coagulation was independent of the concentration. The growth of the coagula is promoted by increasing the SPI concentration and is rate-limiting in the coagulation. PMID- 11853509 TI - Structural characterization of oligosaccharides and polysaccharides from apple juices produced by enzymatic pomace liquefaction. AB - Eight apple pomace liquefaction juices were produced to characterize soluble cell wall material released by the action of pectolytic and cellulolytic enzyme preparations. Very high colloid values from 9.7 to 19.6 g/L were recovered from the juices by ethanol precipitation. The crude polysaccharides consisted mainly of galacturonic acid (49-64 mol %), arabinose (14-23 mol %), galactose (6-15 mol %), and minor amounts of rhamnose, xylose, and glucose. Separation of the polysaccharides by anion-exchange chromatography yielded one neutral, one slightly acidic, and one acidic polymer accounting for 60% of total colloids. Preparative size exclusion chromatography of the acidic fractions resulted in four polymers of different molecular weights and different sugar compositions. Among them, high molecular weight arabinans and rhamnogalacturonans as well as oligomeric fractions consisting of only galacturonic acid could be found. Linkage studies were performed on neutral fractions from anion-exchange chromatography and size exclusion chromatography. They revealed highly branched arabinans, xyloglucans, and mainly type I arabinogalactans. PMID- 11853511 TI - Characterization of major anthocyanins and the color of red-fleshed Budd Blood orange (Citrus sinensis). AB - High-performance liquid chromatography (HPLC) with photodiode array detection was applied for the characterization of anthocyanins in red-fleshed Budd Blood (Citrus sinensis) orange. More than seven anthocyanin pigments were separated within 30 min by using a binary gradient (0.1% H(3)P0(4) in water and 0.1% H(3)PO(4) in acetonitrile) elution on a Prodigy ODS column. Separations by reversed-phase HPLC and semipreparative HPLC on a Prodigy 10-microm ODS Prep column, and acid and alkali hydrolyses were used for identification of anthocyanins. The primary anthocyanins in Budd Blood orange grown in Florida were cyanidin-3-(6"-malonylglucoside) (44.8%) followed by cyanidin-3-glucoside (33.6%). Two other minor pigments were also acylated with malonic acid. Malonated anthocyanins represented the major proportion (>51%) of anthocyanins in Budd Blood orange. Total anthocyanin contents and juice color parameters (CIE L,a,b) were compared with six other Florida-grown blood oranges. PMID- 11853512 TI - Purification and characterization of lipoxygenase from Pleurotus ostreatus. AB - Lipoxygenase was purified homogeneously from cups of Pleurotus ostreatus by Sephacryl S-400 HR gel filtration, Dyematrex Green A affinity, and DEAE-Toyopearl 650M ion-exchange chromatographies. The molecular weight of the enzyme was estimated to be 67,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and 66,000 by gel filtration; the isoelectric point was pH 5.1. The optimum pH and temperature of the enzymatic activity were 8.0 and 25 degrees C, respectively. The enzyme contained non-heme iron, and a thiol group seemed to be involved in its activity. The K(m), V(max), and k(cat) values of the enzyme for linoleic acid were 0.13 mM, 23.4 micromol.min(-1).mg(-1), and 25.7 s(-1), respectively. The enzyme showed high specificity toward linoleic acid. When linoleic acid was incubated with the enzyme, 13-hydroperoxy-9Z,11E octadecadienoic acid was found to be the main oxidative product. PMID- 11853514 TI - Pungency in paprika (Capsicum annuum). 1. Decrease of capsaicinoid content following cellular disruption. AB - The capsaicinoid content in fruits of Capsicum annuum decreased within several days to a level of only 10% of the starting value when cells were disrupted by homogenization. This decrease was not observed in fruits that were carefully cut into halves. The analysis of one half made it possible to determine the reference content at time zero for the second half. A much lower decrease was observed when minced fruits were stored under nitrogen, whereas storage under oxygen resulted in considerable losses of capsaicinoids, indicating oxidative processes as a cause for the decrease of capsaicinoid content. PMID- 11853513 TI - Ability of surfactant micelles to alter the partitioning of phenolic antioxidants in oil-in-water emulsions. AB - In oil-in-water emulsions, the physical location of antioxidants can be an important determinant in their activity. Surfactants can potentially influence the physical location of antioxidants in oil-in-water emulsions by causing solubilization of lipid-soluble antioxidants into the aqueous phase. Excess Brij micelles in an oil-in-water emulsion were found to increase the partitioning of phenolics into the continuous phase with polar antioxidants (propyl gallate) partitioning more than nonpolar antioxidants (butylated hydroxyltoluene). Solubilization of propyl gallate was rapid coming to equilibrium in less than 5 min. Increasing surfactant micelle concentrations from 0.3 to 2.8% increased the solubilization of propyl gallate by 2.3-fold. Solubilization of phenolic antioxidants into the aqueous phase by Brij micelles did not alter the oxidative stability of salmon oil-in-water emulsions, suggesting that surfactant micelles influenced oxidation rates by mechanisms other than antioxidant solubilization. PMID- 11853515 TI - Pungency in paprika (Capsicum annuum). 2. Heterogeneity of capsaicinoid content in individual fruits from one plant. AB - The capsaicinoid content of individual fruits from a single plant harvested at the same time after flowering exhibits a wide range of values with a rather uniform pattern for the ratio of capsaicin, dihydrocapsaicin, and nordihydrocapsaicin. This observation is confirmed by the analysis of fruits from a second and third plant and for several harvest times at different stages of maturity. Competition with lignin-like material, environmental influences, and subcellular distribution may play a role in the synthesis and transformation of capsaicinoids. PMID- 11853516 TI - Identification and quantification of low molecular weight phenolic antioxidants in seeds of evening primrose (Oenothera biennis L.). AB - Crude extracts of evening primrose meal were prepared in 56% (v/v) acetone and separated into six fractions (I-VI) using a Sephadex LH-20 column. Qualitative tests for phenolic and vanillin positive compounds produced positive results for all fractions. Silica gel thin-layer chromatography of fractions III and V allowed the location and isolation of two spots containing moderate to strong antioxidative compounds. High-performance liquid chromatography of the spot isolated from fraction III showed the resolution of two structurally related compounds, whereas that of the spot from fraction V showed the presence of one compound. Nuclear magnetic resonance spectroscopy and electron impact mass spectrometry produced sufficient evidence to identify the isolated compounds as (+)-catechin, (-)-epicatechin, and gallic acid. These compounds accounted for about 10.5 and 1.7% of the dry mass of the crude extracts and meal, respectively. PMID- 11853517 TI - Antiradical activity of water soluble components in common diet vegetables. AB - The antiradical activity of water-soluble components contained in mushrooms (Psalliota campestris), onions (Allium cepa), white cabbage (Brassica oleracea var. alba), and yellow bell peppers (Capsicum annuum) against hydroxyl radicals was tested in a chemical and biological system. The vegetable juices were obtained by centrifugation of a vegetable homogenate processed at 2 degrees C or heated at 102 degrees C. The chemical system consisted of a buffered reaction mixture composed of Fe(III)-EDTA, 2-deoxy-D-ribose, ascorbic acid, and H(2)O(2) generating the hydroxyl radical. The antiradical activity was expressed as an inhibition of deoxyribose degradation. The biological system consisted of IMR32 neuroblastoma cells exposed to H(2)O(2) in the presence or absence of the vegetable juices. Cells were pretreated for either 24 h or 1 h with the vegetable juices, and reduction of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) was used as a cell viability assay. All vegetable juices inhibited the degradation of deoxyribose and increased the viability of H(2)O(2) treated cells. Raw mushroom juice proved to be the most active in both cases. Boiling significantly affected the activity of mushroom juice, but did not change significantly the effect on onions and yellow bell peppers, and partially increased the activity of white cabbage juice. Mushroom antiradical activity was also confirmed by a cytofluorimetric analysis. PMID- 11853518 TI - Deoxyhemoglobin-mediated lipid oxidation in washed fish muscle. AB - Deoxyhemoglobin-mediated lipid oxidation was studied by comparing the pro oxidative activity of anodic and cathodic hemoglobins from trout in a washed cod muscle model system. At pH 6.3, cathodic hemoglobins were nearly fully oxygenated while anodic hemoglobins were poorly oxygenated. Anodic hemoglobins initiated lipid oxidation in washed cod muscle much more rapidly than cathodic hemoglobins, as measured by thiobarbituric acid reactive substances (TBARS) formation. Moreover, anodic hemoglobins appeared to oxidize more rapidly as compared to cathodic hemoglobins in the washed cod muscle model system, as measured by a decrease in redness (a value). A more pronounced pro-oxidative activity of deoxyhemoglobin as compared to oxyhemoglobin was confirmed by accelerated lipid hydroperoxide and TBARS formation in the washed cod muscle model system upon combined addition of anodic hemoglobins and adenosine triphosphate, which is known to lower the oxygenation of anodic hemoglobins at pH 7.2, as compared to only addition of anodic hemoglobins to the washed cod muscle. These studies suggest that deoxyhemoglobin is more pro-oxidative than its oxygenated counterpart at pH values found in postmortem fish muscle. PMID- 11853519 TI - Rapid prediction of gross energy and utilizable energy in cereal food products using near-infrared reflectance spectroscopy. AB - Near-infrared (NIR) spectroscopy has been used in foods for the rapid assessment of several macronutrients; however, little is known about its potential for the evaluation of the utilizable energy of foods. Using NIR reflectance spectra (1104 2494 nm) of ground cereal products (n = 127) and values for energy measured by bomb calorimetry, chemometric models were developed for the prediction of gross energy and available energy of diverse cereal food products. Standard errors of cross-validation for NIR prediction of gross energy (range = 4.05-5.49 kcal/g), energy of samples after adjustment for unutilized protein (range = 3.99-5.38 kcal/g), and energy of samples after adjustment for unutilized protein and insoluble dietary fiber (range = 2.42-5.35 kcal/g) were 0.053, 0.053, and 0.088 kcal/g, respectively, with multiple coefficients of determination of 0.96. Use of the models on independent validation samples (n = 58) gave energy values within the accuracy required for U.S. nutrition labeling legislation. NIR spectroscopy, thus, provides a rapid and accurate method for predicting the energy of diverse cereal foods. PMID- 11853520 TI - Supplementation of plasma with olive oil phenols and extracts: influence on LDL oxidation. AB - Phenols present in olive oil may contribute to the health effects of the Mediterranean lifestyle. Olive oil antioxidants increase the resistance of low density lipoproteins (LDL) against oxidation in vitro, but human intervention studies have failed to demonstrate similar consistent effects. To better mimic the in vivo situation, plasma was incubated with either individual olive oil phenols or olive oil extracts with different phenolic compositions, and LDL was subsequently isolated and challenged for its resistance to oxidation. The results show that the ortho-dihydroxy phenols (hydroxytyrosol and oleuropein-aglycone) are more efficient than their mono-hydroxy counterparts (tyrosol and ligstroside aglycone) in increasing the resistance of LDL to oxidation. However, the concentration of antioxidants required to inhibit LDL oxidation when added to whole plasma was substantially higher as compared to previous data where antioxidants are directly added to isolated LDL. In conclusion, this study supports the hypothesis that extra virgin olive oil phenols protect LDL in plasma against oxidation. The explanation that in vitro studies show protective effects in contrast to the lack of effect in the majority of human studies may be that the dose of the phenols and thus their plasma concentration in humans was too low to influence ex vivo LDL oxidizability. Further studies are required to gain a better understanding of the potential health benefits that extra virgin olive oil may provide. PMID- 11853521 TI - Influence of year and planting date on fatty acid chemistry of high oleic acid and normal peanut genotypes. AB - The effect of an early-, mid-, or late-season planting date on the fatty acid chemistry of four high oleic acid, one mid oleic acid, and five normal oleic acid peanut (Arachis hypogaea L.) genotypes was evaluated over a three year period. Oleic acid was also compared to other fatty acids and to indices of oil quality. High-oleic genotypes included SunOleic 97R, UF98326, UF99621, and 88x1B-OLBC1-6-1 3-1-b2-B with a mean oleic acid content between 77.8 and 82.5%. Florida MDR98, a mid-oleic cultivar, was intermediate in oleic acid chemistry (59.8-68.0%). The normal oil chemistry lines (Georgia Greene, Andru93, Florunner, 86x13A-4-2-3-2-b3 B, and UF97102) had an oleic acid content between 50.0 and 59.0%. The ratio of oleic to linoleic (O/L) was 18:1 to 51:1 for high-oleic lines and 1.7:1 to 3.5:1 for normal genotypes. When analyzed as a split-split plot in time, year had a highly significant effect (P < 0.001) on the eight main fatty acids, iodine value, ratio of unsaturated to saturated fatty acids (U/S), and percentage of saturated fatty acids. Thus, data were analyzed separately by year. Although genotypic effects were highly significant each year, planting date influenced oil chemistry in two of three years. During both 1999 and 2000, 11 of 12 variables were influenced by planting date and by genotype x planting date interactions. Iodine values were approximately 75 for high-oleic lines compared to 90-95 for normal genotypes. The highest correlations occurred for oleic acid (18:1) and linoleic acid (18:2) (r = -0.996) and for oleic and palmitic (16:0) acids (r = 0.959). Oleic acid was also inversely related to iodine value (r = -0.978) and to percentage saturation (r = -0.841). PMID- 11853522 TI - Fungal flora and ochratoxin a production in grapes and musts from france. AB - Eleven samples of grapes and musts used in red table wines were investigated for the occurrence of potential ochratoxin A (OTA)-producing molds. From these samples, 59 filamentous fungi and 2 yeasts were isolated. Among the 30 genera isolated, Deuteromycetes were the most frequent (70%) followed by Ascomycetes (10%). Six of the eleven grapes samples were contaminated by potentially ochratoxinogenic strains (Penicillium chrysogenum and Aspergillus carbonarius). When cultivated in vitro on solid complex media, the 14 strains of A. carbonarius produced OTA. No other species produced OTA under the same conditions. Among must samples, eight of eleven were found to be contaminated by OTA (concentrations from <10 to 461 ng/L). There is a strong correlation between the presence of ochratoxin-producing strains on grapes and OTA in musts. These findings should be connected with the OTA contamination of human blood in these areas and in France. PMID- 11853523 TI - Enantioseparation of vinclozolin by gamma-cyclodextrin-modified micellar electrokinetic chromatography. AB - Cyclodextrin-modified micellar electrokinetic chromatography was applied to the enantioseparation of vinclozolin, which has been used as a fungicide and has an anti-androgenic activity, using gamma-cyclodextrin together with sodium dodecyl sulfate. Factors affecting the chiral resolution and migration time of vinclozolin were studied. The optimum running conditions were found to be 20 mM phosphate-5 mM borate buffer (pH 8.5) containing 50 mM gamma-cyclodextrin and 100 mM sodium dodecyl sulfate with an effective voltage of 20 kV at 20 degrees C using direct detection at 203 nm. Under these conditions, the resolution (Rs) of racemic vinclozolin was approximately 2.1. The sample was concentrated by solid phase extraction and was fractionated by HPLC. The peak area ratio of (+)- and ( )-vinclozolins in wine was found to be 2:3, namely, not racemic, suggesting that degradation rates were different between (+)- and (-)-vinclozolins. The anti androgenic activities of (+)- and (-)-vinclozolins on dihydrotestosterone-induced transcription were also investigated. The anti-androgenic activity of (+) vinclozolin tended to be stronger than that of (-)-vinclozolin, suggesting the possibility that vinclozolin can act as an enantioselective anti-androgen. PMID- 11853524 TI - N-(1-deoxy-D-fructos-1-yl) fumonisin B(1), the initial reaction product of fumonisin B(1) and D-glucose. AB - Incubation of fumonisin B(1) and D-glucose in aqueous solutions resulted in the formation of N-(1-deoxy-D-fructos-1-yl) fumonisin B(1) in addition to the previously reported N-(carboxymethyl) fumonisin B(1). N-(1-Deoxy-D-fructos-1-yl) fumonisin B(1) is the first stable product formed after the Amadori rearrangement of the Schiff base formed by the reaction of the primary amine of fumonisin B(1) and the aldehyde group of D-glucose. N-(1-Deoxy-D-fructos-1-yl) fumonisin B(1) was synthesized by reacting fumonisin B(1) with an excess of D-glucose in methanol and heating for 6 h at 64 degrees C. It was purified using C(18) and strong cation exchange solid-phase extraction cartridges and characterized by nuclear magnetic resonance and liquid chromatography-mass spectrometry. Subsequently, N,N-dimethylformamide was found to be a better reaction solvent, requiring reaction for only 2-3 h at 64 degrees C and eliminating the formation of methyl esters. Alkaline hydrolysis of N-(1-deoxy-D-fructos-1-yl) fumonisin B(1) gave a mixture of hydrolyzed fumonisin B(1) and hydrolyzed N-(carboxymethyl) fumonisin B(1). PMID- 11853525 TI - Self-efficacy in adolescents who have siblings with or without disabilities. AB - Fifty-four adolescents (27 with and 27 without a sibling who had a disability) were contrasted on levels of self-efficacy. Adolescents completed questionnaires measuring self-efficacy, peer competence, and maternal attitudes toward and modeling of prosocial and empathic behavior. None of the adolescent measures differed significantly between those whose siblings did or did not have disabilities. Females evidenced higher levels of self-efficacy than did males, regardless of sibling's disability status. For adolescents with siblings who did not have disabilities, interpersonal competence was significantly related to self efficacy. For adolescents whose siblings had disabilities, interpersonal competence and maternal attitudes and modeling were significantly related to self efficacy. Process-oriented variables associated with self-efficacy were identified and could become targets for intervention efforts to influence this positive set of beliefs in adolescents. PMID- 11853526 TI - Chronological age, receptive vocabulary, and syntax comprehension in children and adolescents with mental retardation. AB - This study was designed to examine the association of CA with syntax and vocabulary comprehension in children and adolescents with mental retardation. It was conducted using the PPVT-R and the Test for Reception of Grammar. The cognitive level of participants was estimated by means of three nonverbal intelligence tests. Results indicated that scores on intelligence tests account for 55% and 29% of the variability of syntax and vocabulary scores, respectively. For the vocabulary, the introduction of CA into the regression equation produced a significant increment of the variance explained (17%). However, its contribution to the variance of the syntax test was not significant. This finding suggests that CA-related experience is linked to on the receptive vocabulary of children and adolescents with mental retardation. PMID- 11853527 TI - Mental retardation and memory for spatial locations. AB - Although memory for spatial location has been frequently investigated with mentally retarded populations, it is not clear that these individuals possess the same spatial memory skills as do their peers without mental retardation. We compared 30 persons with and 30 persons without mental retardation. Following either intentional or incidental learning, participants recalled and then relocated 16 objects on a matrix. The control group recalled more intentionally learned than incidentally learned material. The experimental group performed both tasks better after incidental learning than after intentional learning and scored as highly as the controls on incidental spatial memory. Results suggest that memory for spatial location is an automatic process and, to some extent at least, is not directly affected by the individual's level of intelligence. PMID- 11853528 TI - Sensitivity theory of motivation and psychopathology: an exploratory study. AB - Sensitivity theory, a comprehensive theory of human motivation, states that people with mental retardation are motivated by the same basic motives as are individuals without mental retardation, but to different extents. Reiss and Havercamp, the conceptualizers of this theory, suggested that unusually high or low set points to certain motives may lead to behavioral or psychiatric problems. Here we examined the relationship between psychopathology and motivation in 111 participants with a dual diagnosis and 111 participants with mental retardation only. The former scored higher on 8 of the 15 motives and had more motives above one standard deviation from the mean of the normative sample. Results lend support to the original hypothesis of Reiss and Havercamp. PMID- 11853529 TI - Positive perceptions in families of children with developmental disabilities. AB - There has been increasing recognition of the importance of asking more positive questions about the perceptions and experiences of families of children with developmental disabilities. However, this recognition has not yet had an observable impact on publication trends. Our aims in this paper were to review existing published research on the positive perceptions and experiences of family members and propose a working model for further study of families' positive perceptions. This model is based on the hypothesis that positive perceptions function as strategies that help families adapt to or cope with the experiences of raising a child with disabilities. Throughout the discussion, we identify some questions for future research and address some implications for practice. PMID- 11853530 TI - Food choice in people with Prader-Willi syndrome: quantity and relative preference. AB - Two experiments were conducted to examine differences between people with Prader Willi syndrome and an obese comparison group in choosing a larger quantity of food delivered after a delay (15, 30, or 60 seconds) or a small quantity of food delivered immediately. Individuals with Prader-Willi syndrome selected the larger food quantity. Choices by the obese comparison group tended to be equally divided between both options. Relative food preference influenced choosing the larger, delayed option in both groups. This effect appeared to be larger in the non Prader-Willi syndrome group, but was not statistically significant. These results suggest perceived differences in food quantity may be an important determinant of food choice of people with Prader-Willi syndrome. PMID- 11853531 TI - Quantitative assessments of sensitivity to reinforcement contingencies in mental retardation. AB - Sensitivity to differences in reinforcement contingencies was examined in 6 individuals with mental retardation. A concurrent operants procedure was implemented in the context of a computer game. Participants selected two different types of animated figures displayed concurrently in the left and right portions of a touchscreen monitor. Over a series of conditions, the relative rates or magnitudes of reinforcers following selections of the two options were changed in ratios of 5:1, 3:1, 1:1, 1:3, and 1:5. Sensitivity was quantified by application of the generalized matching equation. Results included individual differences in sensitivity and differential sensitivity to rate and magnitude variation. The results suggest that comprehensive assessments of potential reinforcers may benefit from including tests of delivery parameters. PMID- 11853532 TI - Self-injurious behavior, self-restraint, and compulsive behaviors in Cornelia de Lange syndrome. AB - Researchers have argued that individuals who show self-restraint are more likely to show self-injurious behavior (SIB) that is compulsive. Self-injurious behavior, self-restraint, and compulsive behaviors have been described as features of Cornelia de Lange syndrome. We examined whether individuals with this syndrome displaying SIB and self-restraint exhibit more compulsive behaviors than do those without SIB and self-restraint. Main caregivers of individuals with the syndrome completed questionnaires. A significant association was found between SIB and self-restraint, and those displaying both behaviors displayed significantly more compulsions than did those not exhibiting them. Findings extend the compulsive behavior theory and highlight areas for further research. PMID- 11853533 TI - The transfer of transthyretin and receptor-binding properties from the plasma retinol-binding protein to the epididymal retinoic acid-binding protein. AB - Members of the lipocalin superfamily share a common structural fold, but differ from each other with respect to the molecules with which they interact. They all contain eight beta-strands (A-H) that fold to form a well-defined beta-barrel, which harbours a binding pocket for hydrophobic ligands. These strands are connected by loops that vary in size and structure and make up the closed and open ends of the pocket. In addition to binding ligands, some members of the family interact with other macromolecules, the specificity of which is thought to be associated with the variable loop regions. Here, we have investigated whether the macromolecular-recognition properties can be transferred from one member of the family to another. For this, we chose the prototypical lipocalin, the plasma retinol-binding protein (RBP) and its close structural homologue the epididymal retinoic acid-binding protein (ERABP). RBP exhibits three molecular-recognition properties: it binds to retinol, to transthyretin (TTR) and to a cell-surface receptor. ERABP binds retinoic acid, but whether it interacts with other macromolecules is not known. Here, we show that ERABP does not bind to TTR and the RBP receptor, but when the loops of RBP near the open end of the pocket (L-1, L-2 and L-3, connecting beta-strands A-B, C-D and E-F, respectively) were substituted into the corresponding regions of ERABP, the resulting chimaera acquired the ability to bind TTR and the receptor. L-2 and L-3 were found to be the major determinants of the receptor- and TTR-binding specificities respectively. Thus we demonstrate that lipocalins serve as excellent scaffolds for engineering novel biological functions. PMID- 11853535 TI - Increasing the redox potential of isoform 1 of yeast cytochrome c through the modification of select haem interactions. AB - The oxidation-reduction potential of eukaryotic cytochromes c varies very little from species to species. We have introduced point mutations into isoform 1 of yeast cytochrome c (iso-1-cytochrome c) to selectively engineer a protein with a higher redox potential. Of the ten different mutant proteins generated for the present investigation Y67R, Y67K and W59H were found to be non-functional. Three other mutant proteins, L32M, L32T and T49K, were functional, but too unstable for biophysical studies. Mutant cytochromes c K79S, K79T, Y48H and Y48K were purified and characterized. The Y48K mutant is the only one that exhibits a significant increase of +117 mV in redox potential compared with the wild-type protein while still supporting oxidative phosphorylation in vivo. Low temperature difference spectroscopy confirmed the formation of the holoprotein, while adsorption and CD spectroscopy reveal perturbations in the structure of Y48K iso-1-cytochrome c. PMID- 11853534 TI - The death domain of Rab3 guanine nucleotide exchange protein in GDP/GTP exchange activity in living cells. AB - Rab3 GTPases regulate exocytosis of neurons, endocrine and exocrine cells. In the present paper, we report a system to measure the guanine nucleotide status of Rab3 proteins in living cells. The assay is based on the ability of the Rab3 interacting molecule RIM to extract selectively the GTP-bound form of Rab3. Using this system, we found that approx. 20% of wild-type Rab3A, -B, -C or -D transfected in the insulin-secreting cell line HIT-T15 is in the GTP-bound conformation. The pool of activated Rab3 is decreased under conditions that stimulate exocytosis or by co-expression of the Rab3 GTPase-activating protein. In contrast, co-expression of Mss4 or Rab3-GEP (guanine nucleotide exchange protein) increases by approx. 3-fold the GTP-bound pool of Rab3 isoforms. Rab3 GEP is very similar to MADD, a death domain-containing protein that associates with the type 1 tumour necrosis factor receptor. We observed that the death domain of Rab3-GEP is involved in intramolecular interactions and that deletions or mutations that affect this domain of the protein impair the nucleotide exchange activity towards Rab3. We propose that the death domain of Rab3-GEP acts as a molecular switch and co-ordinates multiple functions of the protein by exchanging its binding partners. PMID- 11853536 TI - Concentrated solutions of salivary MUC5B mucin do not replicate the gel-forming properties of saliva. AB - We have developed a new approach to study the molecular organization of salivary mucus and salivary mucins using confocal fluorescence recovery after photobleaching (confocal-FRAP). MUC5B mucin, its reduced subunit and T-domains were prepared from saliva and fluorescently labelled. The translational self diffusion coefficients were determined up to 3.6 mg/ml by confocal-FRAP. The results suggest that, in solutions of purified MUC5B mucin, at concentrations at which the hydrodynamic domains overlap, the intermolecular interactions are predominantly due to dynamic entanglements, and there was no evidence of specific self-association of MUC5B mucin, or of its subunits, or T-domains. The analysis of the salivary mucus gel also showed no specific interactions with the purified MUC5B components, but it was much less permeable than expected from its MUC5B content. The saliva was completely permeable to microspheres of 207 nm diameter, but showed size-dependent effects on the diffusion of larger microspheres (499 nm and 711 nm diameter). From these analyses the salivary mucus was shown to be both permeable and dynamic, and with the characteristics of a semi-dilute transient network at physiological concentration. Comparison of the results from saliva and purified MUC5B mucin solutions showed that the network properties of saliva were equivalent to a solution of purified MUC5B mucin of 10-20 times higher concentration. This showed that saliva has additional structure and organization not present in the purified MUC5B mucin and suggests there are other interactions and/or components within saliva that combine with MUC5B to produce its complete properties. PMID- 11853537 TI - myo-Inositol hexakisphosphate is a major component of an extracellular structure in the parasitic cestode Echinococcus granulosus. AB - myo-Inositol hexakisphosphate (IP(6)) is an abundant intracellular component of animal cells. In this study we describe the presence of extracellular IP(6) in the hydatid cyst wall (HCW) of the larval stage of the cestode parasite Echinococcus granulosus. The HCW comprises an inner cellular layer and an outer, acellular (laminated) layer up to 2 mm in thickness that protects the parasite from host immune cells. A compound, subsequently identified as IP(6), was detected in and purified from an HCW extract on the basis of its capacity to inhibit complement activation. The identification of the isolated compound was carried out by a combination of NMR, MS and TLC. The majority of IP(6) in the HCW was found in the acellular layer, with only a small fraction of the compound being extracted from cells. In the laminated layer, IP(6) was present in association with calcium, and accounted for up to 15% of the total dry mass of the HCW. IP(6) was not detected in any other structures or stages of the parasite. Our results imply that IP(6) is secreted by the larval stage of the parasite in a polarized fashion towards the interface with the host. This is the first report of the secretion of IP(6), and the possible implications beyond the biology of E. granulosus are discussed. PMID- 11853538 TI - Ecdysone-inducible expression of oncogenic Ha-Ras in NIH 3T3 cells leads to transient nuclear localization of activated extracellular signal-regulated kinase regulated by mitogen-activated protein kinase phosphatase-1. AB - The Ras family of GTP-binding proteins are key transducers of extracellular signals, particularly through the mitogen-activated protein kinase (MAPK) pathway. Constitutively active forms of Ras are found in a variety of tumours, suggesting an important role for this pathway in cancer. Here we report that initial cellular exposure to oncogenic Ras chronically activated the MAPK pathway in the cytoplasm, but transiently activated the same pathway in the nucleus. Nuclear-activated extracellular signal-regulated kinase (ERK) was rapidly dephosphorylated, with consequent short-term activation of the Elk-1 transcription factor and expression of the c-fos gene. Additional experiments suggested that the regulatory mechanism involved requires the calcium-dependent protein phosphotyrosine phosphatase MAPK phosphatase-1 (MKP-1). This is the first report on the ability of Ras, in the absence of growth factors, to transiently activate the MAPK pathway in the nucleus and show an involvement of MKP-1 in nuclear ERK2 regulation. In addition we show that transient activation of the MAPK pathway is sufficient to drive chronic cell-cycle progression. We conclude that, whereas the MAPK pathway is necessary to initiate cellular proliferation and transformation, the transient nature of the MAPK pathway activation suggests the involvement of additional signalling pathway(s) regulated by Ras. PMID- 11853539 TI - Molecular interactions between desmosomal cadherins. AB - Desmocollins (Dscs) and desmogleins (Dsgs) are cell-adhesion molecules involved in the formation of desmosome cell-cell junctions and share structural similarities to classical cadherins such as E-cadherin. In order to identify and provide quantitative information on the types of protein-protein interactions displayed by the type 2 isoforms and investigate the role of Ca(2+) in this process, we have developed an Escherichia coli expression system to generate recombinant proteins containing the first two extracellular domains, namely Dsg2(1-2) and Dsc2(1-2). Analytical ultracentrifugation, chemical cross-linking, CD, fluorescence and BIAcore have been used to provide the first direct evidence of Ca(2+) binding to desmosomal cadherins. These studies suggest that Dsc2(1-2) not only exhibits homophilic interactions in solution, but can also form heterophilic interactions with Dsg2(1-2). The latter, on the other hand, shows much weaker homophilic association. Our results further demonstrate that heterophilic interactions are Ca(2+)-dependent, whereas the Ca(2+)-dependence of homophilic association is less clear. Our data indicate that the functional properties of Dsc2(1-2) are more similar to those of classical cadherins, consistent with the observation that Dsc shares a higher level of sequence homology with classical cadherins than does Dsg. In addition to corroborating the conclusions of previously reported transfection studies which suggest the formation of lateral heterodimers and homodimers, our results also provide direct quantitative information on the strength of these interactions which are essential for understanding the adhesion mechanism. PMID- 11853540 TI - Purification, molecular cloning and heterologous expression of a glutathione S transferase involved in insecticide resistance from the rice brown planthopper, Nilaparvata lugens. AB - A novel glutathione S-transferase (GST)-based pyrethroid resistance mechanism was recently identified in Nilaparvata lugens [Vontas, Small and Hemingway (2001) Biochem. J. 357, 65-72]. To determine the nature of GSTs involved in conferring this resistance, the GSTs from resistant and susceptible strains of N. lugens were partially purified by anion exchange and affinity chromatography. The majority of peroxidase activity, previously correlated with resistance, was confined to the fraction that bound to the affinity column, which was considerably elevated in the resistant insects. A cDNA clone encoding a GST (nlgst1-1) - the first reported GST sequence from Hemiptera with up to 54% deduced amino-acid identity with other insect class I GSTs - was isolated from a pyrethroid-resistant strain. Northern analysis showed that nlgst1-1 was overexpressed in resistant insects. nlgst1-1 was expressed in Escherichia coli, purified and characterized. The ability of the recombinant protein to bind to the S-hexylglutathione affinity matrix, its substrate specificities and its immunological properties confirmed that this GST was one from the elevated subset of N. lugens GSTs. Peroxidase activity of the recombinant nlgst1-1 indicated that it had a role in resistance, through detoxification of lipid peroxidation products induced by pyrethroids. Southern analysis of genomic DNA from the resistant and susceptible strains indicated that GST-based insecticide resistance may be associated with gene amplification in N. lugens. PMID- 11853541 TI - 13C-NMR study of the inhibition of delta-chymotrypsin by a tripeptide-glyoxal inhibitor. AB - A new inhibitor, Z-Ala-Pro-Phe-glyoxal (where Z is benzyloxycarbonyl),has been synthesized and shown to be a competitive inhibitor of delta-chymotrypsin, with a K(i) of 25+/-8 nM at pH 7.0 and 25 degrees C. Z-Ala-Pro-[1-(13)C]Phe-glyoxal and Z-Ala-Pro-[2-(13)C]Phe-glyoxal have been synthesized, and (13)C-NMR has been used to determine how they interact with delta-chymotrypsin. Using Z-Ala-Pro-[2 (13)C]Phe-glyoxal we have detected a signal at 100.7 p.p.m. which we assign to the tetrahedral adduct formed between the hydroxy group of Ser-195 and the (13)C enriched keto-carbon of the inhibitor. This signal is in a pH-dependent slow exchange with a signal at 107.6 p.p.m. which depends on a pK(a) of approximately 4.5, which we assign to oxyanion formation. Thus we are the first to detect an oxyanion pK(a) in a reversible chymotrypsin-inhibitor complex. A smaller titration shift of 100.7 p.p.m. to 103.9 p.p.m. with a pK(a) of approximately 5.3 is also detected due to a rapid exchange process. This pK(a) is also detected with the Z-Ala-Pro-[1-(13)C]Phe-glyoxal inhibitor and gives a larger titration shift of 91.4 p.p.m. to 97.3 p.p.m., which we assign to the ionization of the hydrated aldehyde hydroxy groups of the enzyme-bound inhibitor. Protonation of the oxyanion in the oxyanion hole decreases the binding efficiency of the inhibitor. From this decrease in binding efficiency we estimate that oxyanion binding in the oxyanion hole reduces the oxyanion pK(a) by 1.3 pK(a) units. We calculate that the pK(a)s of the oxyanions of the hemiketal and hydrated aldehyde moieties of the glyoxal inhibitor are both lowered by 6.4-6.9 pK(a) units on binding to chymotrypsin. Therefore we conclude that oxyanion binding in the oxyanion hole has only a minor role in decreasing the oxyanion pK(a). We also investigate how the inhibitor breaks down at alkaline pH, and how it breaks down at neutral pH in the presence of chymotrypsin. PMID- 11853542 TI - Sphingosine 1-phosphate evokes calcium signals in C2C12 myoblasts via Edg3 and Edg5 receptors. AB - Sphingosine 1-phosphate (SPP) is a bioactive lipid that exerts multiple biological effects in a large variety of cell types, acting as either an intracellular messenger or an extracellular ligand coupled to Edg-family receptors (where Edg stands for endothelial differentiation gene). Here we report that in C(2)C(12) myoblasts SPP elicited significant Ca(2+) mobilization. Analysis of the process using a confocal laser-scanning microscope showed that the Ca(2+) response occurred in a high percentage of cells, despite variations in amplitude and kinetics. Quantitative analysis of SPP-induced Ca(2+) transients performed with a spectrophotofluorimeter showed that the rise in Ca(2+) was strictly dependent on availability of extracellular Ca(2+). Cell treatment with pertussis toxin partially prevented the Ca(2+) response induced by SPP, indicating that G(i)-coupled-receptors were involved. Indeed, SPP action was shown to be mediated by agonist-specific Edg receptors. In particular, suramin, an antagonist of the SPP-specific receptor Edg3, as well as down-regulation of Edg3 by cell transfection with antisense oligodeoxyribonucleotides (ODN), significantly reduced agonist-mediated Ca(2+) mobilization. Moreover, an antisense ODN designed to inhibit Edg5 expression also decreased the SPP-induced rise in Ca(2+), although to a lesser extent than that observed by inhibiting Edg3. On the contrary, the SPP response was unaffected in myoblasts loaded with antisense ODN specific for Edg1. Remarkably, the concomitant inhibition of Edg3 and Edg5 receptors abolished the SPP-induced Ca(2+) increase, supporting the notion that Ca(2+) mobilization in C(2)C(12) cells induced by SPP is a receptor mediated process that involves Edg3 and Edg5, but not Edg1. PMID- 11853543 TI - Effect of brefeldin A on heparan sulphate biosynthesis in Madin-Darby canine kidney cells. AB - Brefeldin A (BFA) perturbs the organization of the Golgi apparatus, such that Golgi stack components are fused with the endoplasmic reticulum (ER) and separated from the trans-Golgi network. In many cell types, BFA blocks the secretion of macromolecules but still allows the action of Golgi enzymes in the ER. Treatment of cells with BFA has been reported to inhibit the secretion of heparan sulphate (HS) proteoglycans and alter the structure of their HS components, but the nature of such structural alterations has not been characterized in detail. We analysed the effect of BFA on HS biosynthesis in Madin-Darby canine kidney (MDCK) cells, in which the Golgi complex is more resistant towards BFA than in most other cell types. We found that MDCK cells were able to secrete HS proteoglycans in spite of BFA treatment. However, the secretion of HS was reduced and the secreted HS differed from that produced by untreated cells. In BFA-treated cells, two structurally distinct pools of HS were generated. One pool was similar to HS from control cells, with the exception that the 6-O-sulphation of glucosamine (GlcN) residues was reduced. In contrast, the other pool consisted of largely unmodified N-acetylheparosan polymers with a low (<20%) proportion of N-sulphated GlcN residues but a substantial proportion of N unsubstituted GlcN units, indicating that it had been acted upon by N deacetylases and partly by the N-sulphotransferases, but not by O sulphotransferases. Together, these findings represent a previously unrecognized alteration in HS biosynthesis caused by BFA, and differ dramatically from our previous findings in MDCK cells pertaining to the undersulphation of HS caused by sodium chlorate treatment. PMID- 11853545 TI - Phosphorylation of Xenopus transcription factor IIIA by an oocyte protein kinase CK2. AB - Transcription factor IIIA (TFIIIA), isolated from the cytoplasmic 7 S ribonucleoprotein complex of Xenopus oocytes, is phosphorylated when incubated with [gamma-(32)P]ATP. This modification is due to a trace kinase activity that remains associated with the factor through several steps of purification. The kinase can use either ATP or GTP, and will phosphorylate casein and phosvitin to the exclusion of TFIIIA. The kinase is reactive with a ten-amino-acid peptide that is a specific substrate for protein kinase CK2 (CK2; formerly casein kinase II). In addition, inhibition of phosphorylation by heparin and stimulation by spermidine indicate that the activity can be ascribed to CK2. Phospho amino acid analysis established that serine is the sole phosphoryl acceptor in TFIIIA. There are four consensus sites for CK2 in TFIIIA; all contain serine residues at the putative site of phosphorylation. TFIIIA immunoprecipitated from oocytes, which were incubated with [(32)P]orthophosphate, is also phosphorylated exclusively on serine residues. Only the cyanogen bromide fragment, which was derived from the N terminal end of TFIIIA, is labelled in vivo. A recognition sequence for CK2, located at Ser(16) in the beta-turn of the first zinc-finger domain, is the only protein kinase consensus sequence present in this peptide. Assays in vitro with site-specific mutants of TFIIIA established that Ser(16) is the preferred site of phosphorylation, with some secondary modification at Ser(314). PMID- 11853544 TI - Differentiation regulates interleukin-1beta-induced cyclo-oxygenase-2 in human articular chondrocytes: role of p38 mitogen-activated protein kinase. AB - Chondrocyte dedifferentiation has been noted in osteoarthritic cartilage, but the contribution of this phenomenon is poorly understood. Interleukin (IL)-1beta, the major pro-inflammatory cytokine found in osteoarthritic synovial fluid, induces the dedifferentiation of cultured articular chondrocytes, whereas E-series prostaglandins (PGE) are capable of inducing cell differentiation. Since PGE(2) synthesis is up-regulated by IL-1beta, we addressed the question of whether the state of chondrocyte differentiation may influence the production of IL-1-induced PGE(2) by modulating cyclooxygenase (COX)-2 expression. Immortalized human articular chondrocytes, (tsT/AC62) cultured in monolayer after passage through alginate matrix (alg+) produced 5-fold greater amounts of PGE(2) than continuous monolayer cultures (alg-) after stimulation with IL-1beta. Moreover, IL-1beta induced COX-2 expression at 0.01 ng/ml in (alg+) cells, whereas a 100-fold higher dose of cytokine was necessary for stimulation in (alg-) cells. SB203580, a selective p38 mitogen-activated protein kinase (MAPK) inhibitor, completely abolished the IL-1beta-induced COX-2 mRNA. Overexpression of p38 MAPK induces a COX-2 reporter, whereas overexpression of dominant negative p38 MAPK represses IL 1beta-induced promoter expression. Interestingly, IL-1beta-induced p38 MAPK activity was greatly enhanced in (alg+) compared with (alg-) cells. Our results suggest that differentiated articular chondrocytes are highly responsive to IL 1beta and that p38 MAPK mediates this response by inducing COX-2 gene expression. PMID- 11853546 TI - A novel human small subunit of calpains. AB - Typical calpains are heterodimeric cysteine proteases which have distinct large catalytic subunits (80 kDa) but share a common small regulatory subunit (30 kDa; css1). Here we report the identification, cloning and characterization of a novel human small subunit (css2) encoded by an intronless gene, capns2, located on chromosome 16. This new protein displays 73% sequence identity within the Ca(2+) binding region but lacks two oligo-Gly stretches characteristic of the N-terminal domain of the conventional small subunit. css2 appears to be the functional equivalent of the conventional small subunit in vitro in that it helps the large subunit fold into the active conformation of similar Ca(2+) sensitivity when the two proteins are co-expressed in Escherichia coli. The purification of various chimaeric rat 80 kDa-human css2 constructs, on the other hand, shows that css2 binds the large subunit much more weakly than css1. Further, it does not undergo the autolytic conversion typical of the classical small subunit. The expression of this protein in vivo, as assessed from its appearance in expressed sequence tag clones, is rather limited, making it an example of a tissue-specific, rather than ubiquitous, small subunit. PMID- 11853547 TI - Mutational analysis of the glucagon receptor: similarities with the vasoactive intestinal peptide (VIP)/pituitary adenylate cyclase-activating peptide (PACAP)/secretin receptors for recognition of the ligand's third residue. AB - Receptor recognition by the Asp(3) residues of vasoactive intestinal peptide and secretin requires the presence of a lysine residue close to the second transmembrane helix (TM2)/first extracellular loop junction and an ionic bond with an arginine residue in TM2. We tested whether the glucagon Gln(3) residue recognizes the equivalent positions in its receptor. Our data revealed that the binding and functional properties of the wild-type glucagon receptor and the K188R mutant were not significantly different, whereas all agonists had markedly lower potencies and affinities at the I195K mutated receptor. In contrast, glucagon was less potent and the Asp(3)-, Asn(3)- and Glu(3)-glucagon mutants were more potent and efficient at the double-mutated K188R/I195K receptor. Furthermore, these alterations were selective for position 3 of glucagon, as shown by the functional properties of the mutant Glu(9)- and Lys(15)-glucagon. Our results suggest that although the Gln(3) residue of glucagon did not interact with the equivalent binding pocket as the Asp(3) residue of vasoactive intestinal peptide or secretin, the Asp(3)-glucagon analogue was able to interact with position 188 of the K188R/I195K glucagon receptor. Nevertheless, the Gln(3) side chain of glucagon probably binds very close to this region in the wild-type receptor. PMID- 11853548 TI - Inhibitory effects of acidic phospholipids on the binding of origin-recognition complex to origin DNA. AB - Origin-recognition complex (ORC), a candidate initiator of chromosomal DNA replication in eukaryotes, shares certain biochemical characteristics with DnaA, the initiator of chromosomal DNA replication in prokaryotes. These similarities include origin-specific DNA binding, ATP binding and ATPase activity. DnaA interacts with acidic phospholipids, such as cardiolipin, and its activity is regulated by these phospholipids. In this study, we examined whether Saccharomyces cerevisiae ORC also interacts with phospholipids. Among the various phospholipids tested, ORC was found to bind specifically to cardiolipin. This binding was inhibited by excess concentrations of salts but unaffected by ATP, adenosine 5'-[gamma-thio]triphosphate or the origin DNA. Cardiolipin weakly inhibited the ATP-binding activity of ORC, whereas it strongly inhibited ORC binding to origin DNA. Acidic phospholipids other than cardiolipin (phosphatidylglycerol and phosphatidylinositol) weakly inhibited ORC binding to origin DNA. Furthermore, total phospholipids extracted from yeast nuclear membranes inhibited ORC binding to origin DNA. We consider that phospholipids may modulate initiation of DNA replication in eukaryotes in a similar manner to that found in prokaryotes. PMID- 11853549 TI - Transcriptional regulation of the human manganese superoxide dismutase gene: the role of specificity protein 1 (Sp1) and activating protein-2 (AP-2). AB - Manganese superoxide dismutase (MnSOD) plays an important role in regulating cellular redox conditions. Expression of MnSOD has been shown to protect against damage by oxidative stress and to suppress the malignant phenotype of human cancer cells. We have previously cloned the human MnSOD (SOD2) gene and analysed its 5' proximal promoter, which has been characterized by a lack of a TATA or CAAT box and the presence of multiple GC boxes. To define further the molecular mechanisms for the regulation of MnSOD expression, multiple transcription factor binding motifs containing overlapping specificity protein 1 (Sp1)- and activator protein (AP)-2-binding sites were identified by DNase I footprinting analysis. Functional studies in three cell lines with different levels of Sp1 and AP-2 proteins suggested that the cellular levels of these proteins may differentially regulate transcription via GC-binding motifs in the human SOD2 promoter. Co transfection of an Sp1 expression vector resulted in an increase in the transcription of the promoter-driven reporter gene. In contrast, co-transfection of the AP-2 expression vector caused a decrease in transcription. Direct mutagenesis analysis of Sp1- and AP-2-binding sites showed that Sp1 is essential for transcription of the human SOD2 gene, whereas AP-2 plays a negative role in the transcription. Immunoprecipitation of Sp1 and AP-2 proteins demonstrated that Sp1 interacts with AP-2 in vivo. Two-hybrid analysis revealed that interaction between Sp1 and AP-2 plays both a positive and negative role in the transcription of the reporter gene in vivo. Taken together, our data indicate that AP-2 down regulates transcription of the human SOD2 gene via its interaction with Sp1 within the promoter region. These findings, coupled with our previous observation that several cancer cell lines have mutations in the promoter region of the human MnSOD gene, which lead to an increase in an AP-2-binding site and a decrease in the promoter activity, signal the importance of understanding the promoter structure and the regulation of the human SOD2 gene by Sp1 and AP-2. PMID- 11853550 TI - A natural protective mechanism against hyperglycaemia in vascular endothelial and smooth-muscle cells: role of glucose and 12-hydroxyeicosatetraenoic acid. AB - Bovine aortic endothelial and smooth-muscle cells down-regulate the rate of glucose transport in the face of hyperglycaemia, thus providing protection against deleterious effects of increased intracellular glucose levels. When exposed to high glucose concentrations these cells reduced the mRNA and protein content of their typical glucose transporter, GLUT-1, as well as its plasma membrane abundance. Inhibition of the lipoxygenase (LO) pathway, and particularly 12-LO, reversed this glucose-induced down-regulatory process and restored the rate of hexose transport to the level seen in vascular cells exposed to normal glucose levels. This reversal was accompanied by increased levels of GLUT-1 mRNA and protein, as well as of its plasma-membrane content. Exposure of the vascular cells to elevated glucose concentrations increased by 2-3-fold the levels of cell associated and secreted 12-hydroxyeicosatetraenoic acid (12-HETE), the product of 12-LO. Inhibition of 15- and 5-LO, cyclo-oxygenases 1 and 2, and eicosanoid producing cytochrome P450 did not modify the hexose-transport system in vascular cells. These results suggest a role for HETEs in the autoregulation of hexose transport in vascular cells. 8-Iso prostaglandin F(2alpha), a non-enzymic oxidation product of arachidonic acid, had no effect on the hexose-transport system in vascular cells exposed to hyperglycaemic conditions. Taken together, these findings show that hyperglycaemia increases the production rate of 12-HETE, which in turn mediates the down-regulation of GLUT-1 expression and the glucose transport system in vascular endothelial and smooth-muscle cells. PMID- 11853551 TI - Measurement of ferrochelatase activity using a novel assay suggests that plastids are the major site of haem biosynthesis in both photosynthetic and non photosynthetic cells of pea (Pisum sativum L.). AB - Ferrochelatase is the terminal enzyme of haem biosynthesis, catalysing the insertion of ferrous iron into the macrocycle of protoporphyrin IX, the last common intermediate of haem and chlorophyll synthesis. Its activity has been reported in both plastids and mitochondria of higher plants, but the relative amounts of the enzyme in the two organelles are unknown. Ferrochelatase is difficult to assay since ferrous iron requires strict anaerobic conditions to prevent oxidation, and in photosynthetic tissues chlorophyll interferes with the quantification of the product. Accordingly, we developed a sensitive fluorimetric assay for ferrochelatase that employs Co(2+) and deuteroporphyrin in place of the natural substrates, and measures the decrease in deuteroporphyrin fluorescence. A hexane-extraction step to remove chlorophyll is included for green tissue. The assay is linear over a range of chloroplast protein concentrations, with an average specific activity of 0.68 nmol x min(-1) x mg of protein(-1), the highest yet reported. The corresponding value for mitochondria is 0.19 nmol x min(-1) x mg of protein(-1). The enzyme is inhibited by N-methylprotoporphyrin, with an estimated IC(50) value of approximately 1 nM. Using this assay we have quantified ferrochelatase activity in plastids and mitochondria from green pea leaves, etiolated pea leaves and pea roots to determine the relative amounts in the two organelles. We found that, in all three tissues, greater than 90% of the activity was associated with plastids, but ferrochelatase was reproducibly detected in mitochondria, at levels greater than the contaminating plastid marker enzyme, and was latent. Our results indicate that plastids are the major site of haem biosynthesis in higher plant cells, but that mitochondria also have the capacity for haem production. PMID- 11853552 TI - Interaction of syncollin with GP-2, the major membrane protein of pancreatic zymogen granules, and association with lipid microdomains. AB - Syncollin, a novel pancreatic zymogen granule protein, is present on the luminal side of the granule membrane. To address the function of syncollin, we searched for putative binding partners. Cross-linking experiments with purified syncollin, and granule content and membrane proteins revealed a direct interaction between syncollin and GP-2, a major glycosylphosphatidylinositol (GPI)-anchored membrane glycoprotein. An interaction was also observed when cross-linking was performed with recombinant GP-2. In addition, syncollin could be cross-linked to itself, supporting the suggestion that it exists as a homo-oligomer. Cleavage of the GPI anchor of GP-2 by treatment of granule membranes with phosphatidylinositol specific phospholipase C had no effect on the membrane attachment of syncollin, indicating that it is not mediated exclusively via an interaction with GP-2. Syncollin was found to be associated with detergent-insoluble cholesterol/glycolipid-enriched complexes. These complexes floated to the lighter fractions of sucrose-density gradients and also contained GP-2, the lectin ZG16p, sulphated matrix proteoglycans and the soluble N-ethylmaleimide-sensitive fusion protein attachment protein receptors (SNAREs) syntaxin 3 and synaptobrevin 2. Our results indicate that membrane-associated syncollin is a component of lipid rafts, where it interacts both with GP-2 and membrane lipids. We suggest that the syncollin-GP-2 complex might play a role in signal transduction across the granule membrane. PMID- 11853553 TI - Two mutations in troponin I that cause hypertrophic cardiomyopathy have contrasting effects on cardiac muscle contractility. AB - We investigated the effects of two mutations in human cardiac troponin I, Arg(145)-->Gly and Gly(203)-->Ser, that are reported to cause familial hypertrophic cardiomyopathy. Mutant and wild-type troponin I, overexpressed in Escherichia coli, were used to reconstitute troponin complexes in vanadate treated guinea pig cardiac trabeculae skinned fibres, and thin filaments were reconstituted with human cardiac troponin and tropomyosin along with rabbit skeletal muscle actin for in vitro motility and actomyosin ATPase assays. Troponin containing the Arg(145)-->Gly mutation inhibited force in skinned trabeculae less than did the wild-type, and had almost no inhibitory function in the in vitro motility assay. There was an enhanced inhibitory function with mixtures of 10-30% [Gly(145)]troponin I with the wild-type protein. Skinned trabeculae reconstituted with troponin I containing the Gly(203)-->Ser mutation and troponin C produced less Ca(2+)-activated force (64+/-8% of wild-type) and demonstrated lower Ca(2+) sensitivity [Delta(p)Ca(50) (log of the Ca(2+) concentration that gave 50% of maximal activation) 0.25 unit (P<0.05)] compared with wild-type troponin I, but thin filaments containing [Ser(203)]-troponin I were indistinguishable from those containing the wild-type protein in in vitro motility and ATPase assays. Thus these two mutations each result in hypertrophic cardiomyopathy, but have opposite effects on the overall contractility of the muscle in the systems we investigated, indicating either that we have not yet identified the relevant alteration in contractility for the Gly(203)->Ser mutation, or that the disease does not result directly from any particular alteration in contractility. PMID- 11853554 TI - Biochemical analysis of DnaA protein with mutations in both Arg328 and Lys372. AB - The DnaA protein is the initiator of chromosomal DNA replication in Escherichia coli. Acidic phospholipids decrease its affinity for adenine nucleotides, and re activate the ADP-bound form to the ATP-bound form. We have previously reported that two mutant forms, DnaAR328E and DnaAK372E, have decreased affinity for cardiolipin (CL). In the present study, we constructed a mutant DnaA protein, DnaA435, with both R328E and K372E, and compared its biochemical characteristics with those of DnaAR328E and DnaAK372E. DnaA435 could bind to oriC DNA, but did not bind ATP or ADP. In DnaA435, compared with DnaAR328E and DnaAK372E, CL caused less inhibition of oriC DNA binding, suggesting that amino acids R328 and K372 are involved in the interaction of DnaA with acidic phospholipids. DnaA435 could initiate DNA synthesis on oriC both in vivo and in vitro. Based on these results, we propose that ATP activates DnaA protein by changing its higher order structure around R328 and K372. PMID- 11853555 TI - Nitric oxide inactivation in brain by a novel O2-dependent mechanism resulting in the formation of nitrate ions. AB - In order for nitric oxide (NO) to function as a biological messenger it has to be inactivated, but little is known of how this is achieved. In cells from the brain, we have recently shown the existence of a powerful NO sink that 'shapes' NO signals for targeting its receptor, soluble guanylate cyclase, whilst simultaneously preventing NO rising to toxic concentrations [Griffiths and Garthwaite (2001) J. Physiol. (Cambridge, U.K.) 536, 855-862]. In the present study, the properties of this sink were investigated further. Inactivation of NO was preserved in rat brain homogenates. In both cerebellar cell suspensions and brain homogenates, NO inactivation required O(2) and, from measurements in homogenates, the principal end-product was NO(-)(3), which is also the main product of endogenously formed NO in vivo. Direct chemical reaction with O(2), superoxide anions or haemoglobin was not responsible. Consumption of NO was, however, inhibited by heat or protease treatment. Pharmacological tests were negative for several candidate enzymes, namely cytochrome c oxidase, H(2)O(2) dependent haem peroxidases, prostaglandin H synthase, 12/15-lipoxygenase and a flavohaemoglobin-like NO dioxygenase. The capacity of the NO sink in cells was limited because regeneration of the activity was slow (2 h). It is concluded that NO is consumed in the brain through a novel protein, ultimately forming NO(-)(3), and that the slow regeneration of the activity provides a scenario for NO to become toxic. PMID- 11853556 TI - The mechanism of aggrecan release from cartilage differs with tissue origin and the agent used to stimulate catabolism. AB - The mechanisms of aggrecan degradation in adult human articular, adult bovine nasal and fetal bovine epiphyseal cartilage in response to either interleukin 1beta (IL-1beta) or retinoic acid were compared using an explant culture system. Bovine nasal cartilage cultured with either IL-1beta or retinoic acid exhibited significant release of glycosaminoglycan (GAG). For both factors, aggrecan proteolysis occurred predominantly at the 'aggrecanase' site, with no evidence for the action of matrix metalloproteinases, and resulted in the appearance of the corresponding G1 fragment in tissue extracts and in culture media. In human cartilage, little effect of IL-1beta was seen, but abundant release of GAG occurred in the presence of retinoic acid, with evidence of aggrecanase action. Treatment of fetal epiphyseal cartilage with retinoic acid resulted in significant GAG release, whereas treatment with IL-1beta did not. In the retinoic acid-treated tissue, however, no evidence for the cleavage of aggrecan in the interglobular region was apparent. Thus, in the fetal system, agents in addition to aggrecanase and matrix metalloproteinases appear to be active. Taken together, these data demonstrate that the pathways utilized for aggrecan catabolism may vary between different cartilages for a given stimulatory agent, and that, for a given tissue, different factors may elicit aggrecan release via different pathways. PMID- 11853557 TI - Hyaluronate degradation as an alternative mechanism for proteoglycan release from cartilage during interleukin-1beta-stimulated catabolism. AB - Data presented previously suggest that release of components of the cartilage matrix, in response to catabolic agents, cannot be accounted for by proteolytic mechanisms alone. In the present study, the release of glycosaminoglycan containing components from bovine nasal cartilage cultured in the presence of interleukin-1beta, and from bovine nasal, fetal bovine epiphyseal and adult human articular cartilage cultured in the presence of retinoic acid, was accompanied by the loss of link protein and hyaluronate into the culture medium. Chromatographic analysis of the released hyaluronate showed it to be markedly reduced in size relative to that extracted from the corresponding tissue. It is proposed that, under stimulation by catabolic agents, two independent, but concurrent, mechanisms act to promote the release of aggrecan from the cartilage matrix. First, proteolytic cleavage of the aggrecan core protein results in the production of glycosaminoglycan-containing fragments that are free to diffuse from the tissue. Secondly, cleavage of hyaluronate renders portions of the proteoglycan aggregate small enough so that complexes of aggrecan (or fragments containing its G1 domain) and link protein are released from the tissue. It is likely that both mechanisms contribute to cartilage metabolism in normal physiology and pathology. PMID- 11853558 TI - Identification and characterization of four novel phosphorylation sites (Ser31, Ser325, Thr336 and Thr366) on LKB1/STK11, the protein kinase mutated in Peutz Jeghers cancer syndrome. AB - Peutz-Jeghers syndrome is an inherited cancer syndrome, which results in a greatly increased risk of developing tumours in those affected. The causative gene encodes a nuclear-localized protein kinase, termed LKB1, which is predicted to function as a tumour suppressor. The mechanism by which LKB1 is regulated in cells is not known, and nor have any of its physiological substrates been identified. Recent studies have demonstrated that LKB1 is phosphorylated in cells. As a first step towards identifying the roles that phosphorylation of LKB1 play, we have mapped the residues that are phosphorylated in human embryonic kidney (HEK)-293 cells, as well as the major in vitro autophosphorylation sites. We demonstrate that LKB1 expressed in HEK-293 cells, in addition to being phosphorylated at Ser(431), a previously characterized phosphorylation site, is also phosphorylated at Ser(31), Ser(325) and Thr(366). Incubation of wild-type LKB1, but not a catalytically inactive mutant, with manganese-ATP in vitro resulted in the phosphorylation of LKB1 at Thr(336) as well as at Thr(366). We were unable to detect autophosphorylation at Thr(189), a site previously claimed to be an LKB1 autophosphorylation site. A catalytically inactive mutant of LKB1 was phosphorylated at Ser(31) and Ser(325) in HEK-293 cells to the same extent as the wild-type enzyme, indicating that LKB1 does not phosphorylate itself at these residues. We show that phosphorylation of LKB1 does not directly affect its nuclear localization or its catalytic activity in vitro, but that its phosphorylation at Thr(336), and perhaps to a lesser extent at Thr(366), inhibits LKB1 from suppressing cell growth. PMID- 11853559 TI - The unfolded protein response in a dolichyl phosphate mannose-deficient Chinese hamster ovary cell line points out the key role of a demannosylation step in the quality-control mechanism of N-glycoproteins. AB - The CHO (Chinese hamster ovary) glycosylation mutant cell line, B3F7, transfers the truncated glycan Glc(3)Man(5)GlcNAc(2) on to nascent proteins. After deglucosylation, the resulting Man(5)GlcNAc(2) glycan is subjected to two reciprocal enzymic processes: the action of an endoplasmic-reticulum (ER) kifunensine-sensitive alpha1,2-mannosidase activity to yield a Man(4)GlcNAc(2) glycan, and the reglucosylation involved in the quality-control system which ensures that only correctly folded glycoproteins leave the ER. We show that the recombinant secreted alkaline phosphatase (SeAP) produced in stably transfected B3F7 cells, is co-immunoprecipitated with the GRP78 (glucose-regulated protein 78), a protein marker of the unfolded protein response (UPR). The level of GRP78 transcription has been evaluated by reverse transcription-PCR (RT-PCR) and we demonstrate that B3F7 cells present a constitutively higher level of UPR in the absence of inductors, compared with Pro(-5) cells. Interestingly, a decrease was observed in the UPR and an increase in SeAP secretion in the kifunensine-treated B3F7 cells. Altogether, these data highlight the relationships between the glycan structure, the quality control system and the UPR. Moreover, they support the idea that a specific demannosylation step is a key event of the glycoprotein quality control in B3F7 cells. PMID- 11853560 TI - Evidence for the direct interaction between calmodulin and the human epidermal growth factor receptor. AB - Previous work from our laboratory has demonstrated that the Ca(2+)-calmodulin complex inhibits the intrinsic tyrosine kinase activity of the epidermal growth factor receptor (EGFR), and that the receptor can be isolated by Ca(2+)-dependent calmodulin-affinity chromatography [San Jose, Bengurija, Geller and Villalobo (1992) J. Biol. Chem. 267, 15237-15245]. Moreover, we have demonstrated that the cytosolic juxtamembrane region of the human receptor (residues 645-660) binds calmodulin in a Ca(2+)-dependent manner when this segment forms part of a recombinant fusion protein [Martijn-Nieto and Villalobo (1998) Biochemistry 37, 227-236]. However, demonstration of the direct interaction between calmodulin and the whole receptor has remained elusive. In this work, we show that calmodulin, in the presence of Ca(2+), forms part of a high-molecular-mass complex built upon covalent cross-linkage of the human EGFR immunoprecipitated from two cell lines overexpressing this receptor. Although several calmodulin-binding proteins co immunoprecipitated with the EGFR, suggesting that they interact with the receptor, we demonstrated using overlay techniques that biotinylated calmodulin binds directly to the receptor in a Ca(2+)-dependent manner without the mediation of any adaptor calmodulin-binding protein. Calmodulin binds to the EGFR with an apparent dissociation constant (K'(d)) of approx. 0.2-0.3 microM. Treatment of cells with epidermal growth factor, or with inhibitors of protein kinase C and calmodulin-dependent protein kinase II, or treatment of the immunoprecipitated receptor with alkaline phosphatase, does not significantly affect the binding of biotinylated calmodulin to the receptor. PMID- 11853562 TI - Detection of cross-transmission of multiresistant Gram-negative bacilli and Staphylococcus aureus in adult intensive care units by routine typing of clinical isolates. PMID- 11853563 TI - Bacteria and yeasts in the treatment of acute and chronic infectious diarrhea. Part II: Yeasts. PMID- 11853564 TI - Individual value of each of the Duke criteria for the diagnosis of infective endocarditis. AB - OBJECTIVES: To assess the value of each of the Duke criteria for the diagnosis of infective endocarditis (IE). METHODS: Detailed review was done of charts of all cases discharged with the diagnosis of IE, and classification as 'definite', 'possible' and 'rejected' cases was made according to the Duke criteria. The diagnostic impact of each criterion was assessed by reclassifying each 'definite' case after subtraction of each individual criterion and by reclassifying each 'possible' and 'rejected' case after addition of each individual criterion. RESULTS: From 1983 to 1993, 179 cases were identified in the databases of two hospitals. When the Duke criteria were applied, 124 (6967%) were classed as 'definite', 43 (2466%) as 'possible' and 12 (763%) as 'rejected' cases. Of the 67 pathologically proven cases, 52 (78610%) were 'definite' cases when the criteria were applied before pathology. If the major microbiological criterion is subtracted, 53% (69%) of the 'definite' cases become 'possible' or 'rejected'. When the echocardiographic criterion is subtracted, 34% (68%) of the 'definite' cases become possible or rejected. Among minor criteria, fever and predisposition, contributing to the classification of respectively 31% (68%) and 27% (68%) of the 'definite' cases, were the most powerful. On the other hand, the minor microbiological criterion and immunologic phenomena were responsible for the classification of only 2% (62%) and 6% (64%) respectively, of the 'definite' cases. CONCLUSIONS: Depending on the criterion examined, 47-98% of the 'definite' cases of IE would remain 'definite' if this particular criterion were absent. The major microbiological criterion had the highest relative importance. In this retrospective study, in which only 32 (18%) patients had a transesophageal echocardiogram, the echocardiogram contributed to 15% (66%) of the 'definite' cases according to the major criterion and to 19% (66%) according to the minor criterion. This study illustrates that the degree of certainty of the diagnosis of IE often depends on the presence/absence of only one criterion. PMID- 11853561 TI - Dehydroascorbic acid uptake by coronary artery smooth muscle: effect of intracellular acidification. AB - Dehydroascorbic acid (DHAA) enters cells via Na(+)-independent glucose transporters (GLUT) and is converted to ascorbate. However, we found that Na(+) removal inhibited [(14)C]DHAA uptake by smooth-muscle cells cultured from pig coronary artery. The uptake was examined for 2-12 min at 10-200 microM DHAA in either the presence of 134 mM Na(+) or in its absence (N-methyl D-glucamine, choline or sucrose replaced Na(+)). This inhibition of DHAA uptake by Na(+) removal was paradoxical because it was inhibited by 2-deoxyglucose and cytochalasin B, as expected of transport via the GLUT pathway. We tested the hypothesis that this paradox resulted from an inefficient intracellular reduction of [(14)C]DHAA into [(14)C]ascorbate upon intracellular acidosis caused by the Na(+) removal. Consistent with this hypothesis: (i) the Na(+)/H(+)-exchange inhibitors ethylisopropyl amiloride and cariporide also decreased the uptake, (ii) Na(+) removal and Na(+)/H(+)-exchange inhibitors lowered cytosolic pH, with the decrease being larger in 12 min than in 2 min, and (iii) less of the cellular (14)C was present as ascorbate (determined by HPLC) in cells in Na(+)-free buffer than in those in Na(+)-containing buffer. This inability to obtain ascorbate from extracellular DHAA may be detrimental to the coronary artery under hypoxia induced acidosis during ischaemia/reperfusion. PMID- 11853565 TI - Antimicrobial susceptibility of non-Bacteroides fragilis group anaerobic Gram negative bacilli in Europe. AB - OBJECTIVE: To investigate the in vitro activity of a range of anti-anaerobe antimicrobials against non-Bacteroides fragilis group anaerobic Gram-negative bacilli isolated in Europe. METHODS: Isolates from 15 laboratories in 13 countries were identified by conventional methods. The MICs of 20 antibiotics were determined by an agar dilution method. RESULTS: There were 488 Prevotella spp., 174 fusobacteria, 69 Porphyromonas spp., 33 Bacteroides spp., 28 Bilophila wadsworthia and 16 Campylobacter spp. isolates, one Sutterella wadsworthensis isolate and four unidentified isolates. Penicillin resistance (and diminished susceptibility to piperacillin) was most common in Prevotella spp. and Bilophila wadsworthia but was also seen in many other species. All isolates, except three of Bilophila wadsworthia, were susceptible to amoxycillin/clavulanate. Most isolates were susceptible to cefoxitin (except Bilophila wadsworthia) and all were susceptible to the carbapenems. Clinafloxacin was the most active quinolone, followed by trovafloxacin and then sparfloxacin. Most fusobacteria were inherently resistant to the macrolides, as expected, but resistance to macrolides and a ketolide in other species was uncommon. Most Fusobacterium varium isolates were resistant to clindamycin, but resistance to clindamycin in all other species was rare. Tetracycline resistance was common but this did not affect the glycylcyclines. There was one isolate of Bacteroides putredinis resistant to chloramphenicol, and three isolates, a Bacteroides ureolyticus isolate, the Sutterella wadsworthensis isolate and one of the unnamed isolates, were metronidazole resistant. Rifampicin was active against most Prevotella and Porphyromonas spp., but not against many other genera. CONCLUSIONS: Penicillin resistance has increased in Europe among non-Bacteroides fragilis anaerobic Gram negative bacilli, much of it due to beta-lactamase. Acquired resistance to other beta-lactams, macrolides and rifampicin has not significantly increased, and chloramphenicol and metronidazole are unaffected. However, resistance to tetracycline is common. The new compounds, a ketolide (HMR 3647), the glycylcyclines and clinafloxacin, are highly active. PMID- 11853566 TI - Evaluation of the Rapid ID 32 Strep system. AB - OBJECTIVE: To evaluate the performance of the Rapid ID 32 Strep system in the hands of clinical microbiologists without expert knowledge of streptococci or enterococci. METHODS: One hundred and twenty-two strains of streptococci and enterococci conventionally identified in a reference laboratory were sent under code numbers to a clinical microbiology laboratory and identified with the Rapid ID 32 Strep system. RESULTS: Regardless of whether automatic reading and identification or visual reading with identification using tables were done, 75 77% of the 122 examined strains were correctly identified, 7% were misidentified and 16-18% could not be identified with certainty to the species level. The system correctly identified the majority of the examined pyogenic streptococci and enterococci, but only two-thirds of the viridans streptococcal strains. CONCLUSIONS: In a routine laboratory, the Rapid ID 32 Strep system can be used to give a rapid preliminary identification of streptococci and enterococci, but with viridans streptococci one would have to accept a certain risk of mis identification. The assay can, however, be used to biotype viridans streptococci in order to attempt to establish identity between separate isolates, e.g. from blood in patients suspected of having endocarditis. PMID- 11853567 TI - An interprovincial external quality assessment of the ability of Canadian laboratories to detect the vancomycin and penicillin resistance of Enterococcus faecium D366. AB - OBJECTIVE: To evaluate the ability of Canadian laboratories to identify enterococci and detect low-level resistance to penicillin, ampicillin and vancomycin in five provinces and two territories by two external quality assessment schemes. METHODS: Enterococcus faecium, strain D366, with minimum inhibitory concentrations for vancomycin and penicillin of 32 and 16 mg/L respectively, was distributed during a routine proficiency survey. Laboratories were required to culture and identify the isolate and to test antimicrobial susceptibility. Participants were assessed against consensus reference values. RESULTS: Three hundred and sixty-four hospital, commercial and public-health laboratories participated, using their established procedures for patient samples. The isolate was identified to the species level by 222 (61%) laboratories and to the genus level by a further 98 participants. Forty-four failed to meet the expected standard. Vancomycin resistance was detected by 94%. Those reporting a falsely susceptible result used disk diffusion testing. Penicillin resistance was noted by 250 of 258 laboratories reporting on this agent. An incorrect ampicillin-susceptible finding was reported by 62 of 147 laboratories using automated microdilution or agar dilution methods. CONCLUSIONS: Most laboratories identified the isolate to an appropriate level. Detection of low-level vancomycin and penicillin resistance was achieved by the majority. Ampicillin resistance was less readily detected. PMID- 11853568 TI - Factors associated with survival in human immunodeficiency virus-positive patients and disseminated Mycobacterium avium complex infection. PMID- 11853569 TI - Antimicrobial susceptibility of coagulase-negative staphylococci isolated between 1991 and 1996 from a German university hospital. PMID- 11853570 TI - Interaction of insulin and Burkholderia cepacia. PMID- 11853571 TI - Urinary tract infection associated with Comamonas acidovorans. PMID- 11853572 TI - High-level cephalosporinase-producing Enterobacter cloacae meningitis in a newborn. PMID- 11853573 TI - Mollaret meningitis with orolabial herpes and large lysed ghost cells. PMID- 11853574 TI - [A clinical study on fresh amniotic membrane transplantation for treatment of severe ocular surface disorders at acute inflammatory and cicatricial stage]. AB - OBJECTIVE: To investigate the safety and evaluate the effect of fresh amniotic membrane transplantation to reconstruct the ocular surface with severe inflammation and scarring. METHOD: Amniotic membrane transplantation (38 cases, 46 eyes), or combined with either limbal transplantation (9 cases, 9 eyes) or lamellar keratoplasty (8 cases, 8 eyes) was performed on ocular surface burns (5 cases, 6 eyes), recurrent Mooren's ulcer (8 cases, 8 eyes) and severe symblepharon (38 cases, 46 eyes). All the patients were followed-up for 6 - 18 months (mean, 11 months). Impression cytology was examined in 21 eyes with normal tear secretion post-operatively. RESULTS: No acute rejection was observed after fresh amniotic membrane transplantation. No progressive melt and perforation were seen in five out of six eyes with burns in reconstructed ocular surfaces. Neovascularization, pseudopterygium and iris atrophy were not found during the follow-up. Visual acuity was improved in various degrees. Corneal ulceration was not noted in 8 cases (8 eyes) with frequently recurrent Mooren's ulcer. Ocular surface reconstruction was successful in 46 of 49 eyes with severe symblepharon. Symblepharon occurred in the remaining three eyes but much less than pre transplantation in degree. Amniotic epithelia have been examined in about three months on the transplanted eyes with normal tear secretion. CONCLUSION: Fresh amniotic membrane can be used as a graft to reconstruct ocular surface. Such transplantation can effectively reduce neovascularization, fibrosis and inflammation. Our data indicated that complete removal of pathological tissues and perfect fixation of amniotic membrane graft are crucial steps in the reconstruction of ocular surface. PMID- 11853575 TI - [Evaluation of the results of therapeutic lamellar keratoplasty and penetrating keratoplasty for fungal corneal ulcer]. AB - OBJECTIVE: To evaluate the effects of therapeutic lamellar and penetrating keratoplasty for fungal corneal ulcer. METHODS: Donor corneas preserved in pure glycerine or water-free calcium chloride were used in corneal transplantation for fungal corneal ulcers that were poorly responsive to the anti-fungal medical treatment, 5% iodine was used in lamellar keratoplasty to kill the fungi in the recipient bed, and diluted or original solution of fluconazole was used in penetrating keratoplasty to wash the anterior chamber. RESULTS: In the lamellar keratoplasty group, there were 23 cases; 18 cases were cured with one surgery and 3 cases were cured after second surgery, the success rate being 91.3%. There were 2 cases with recurrence of corneal inflammation, and the lesion was controlled by penetrating keratoplasty. The penetrating keratoplasty group included 31 cases, of them, 27 cases were cured with one surgery and 4 eyes were enucleated due to recurrent corneal inflammation or refractive glaucoma. In the total 54 cases, the inflammation was controlled in 50 cases after therapeutic corneal transplantation, success rate being 92.6%. CONCLUSIONS: For those fungal corneal ulcers that are poorly responsive to anti-fungus eye drops, it is recommended to perform lamellar keratoplasty if the ulcer is located in the anterior 1/2 stroma, the key points of surgery are to remove the affected tissue completely and cauterize the recipient bed with 5% iodine. For deep ulcers or nearly perforative or perforated ulcers, diluted or original solution of diflucan is used to wash the anterior chamber during penetrating keratoplasty, that can help to improve the success rate. PMID- 11853576 TI - [A report on investigation and clinical application of corneal storage media]. AB - OBJECTIVE: To develop intermediate term storage media suitable for Chinese eye banks. METHODS: Corneal buttons of rabbits were stored in DX solution freshly prepared. After storage for several days, the survival rate of the corneal endothelial cells was examined, enzymohistochemical staining and ultrastructural examinations were carried out for the buttons. Thirty-seven human corneas stored in DX solution for 3 - 11 days were used for corneal transplantation, clinically. RESULTS: There was no significant difference between rabbit corneal buttons stored in DX sol. and in Optisol in any indices studied. 37 grafts were all transparent in 1 week after transplantation, and the mean endothelial cell density was (2,204.56 +/- 689.56) cells/mm(2). CONCLUSION: As a safe and efficient intermediate term (about 1 week) storage medium, DX sol. is appropriate for Chinese eye banks. PMID- 11853577 TI - [Effect of transforming growth factor - beta1 on expression of CD69(+) and CD25(+) T cell induced by allo - corneal epithelial cells in vitro]. AB - OBJECTIVE: To study the effect of transforming growth factor - beta(1) (TGF beta1) on changes of CD(69)(+) and CD(25)(+) T cell population induced by allo - corneal epithelial cells in culture. METHOD: The expression of CD(69)(+) and CD(25)(+) in peripheral T cells was identified by using indirect immune fluorescence flow cytometry analysis system. RESULT: The expression of CD(69)(+) and CD(25)(+) in peripheral T cells was significantly increased after peripheral blood had been cultured with allo - corneal epithelial cells for 3 hours However, the expression of CD(69)(+) was significantly decreased while TGF - beta(1) was added to the culture. CONCLUSION: TGF - beta(1) significantly inhibits the expression of CD(69)(+) and CD(25)(+) in peripheral T cells induced by allo corneal epithelial cells in vitro. PMID- 11853578 TI - [An experimental study of immune mediating action of T-lymphocyte subsets on recurrent herpetic stromal keratitis]. AB - OBJECTIVE: To investigate the role of T-lymphocyte subsets in the recurrent herpetic stromal keratitis (SK). METHODS: In a recurrent SK NIH mouse model, rat monoclonal antibodies were used to selectively deplete in vivo the CD(+)(4) and CD(+)(8) T-lymphocytes in mice, and the effects of the subsets on recurrent herpetic SK were evaluated. RESULTS: As measured by flow cytometry, in the mice treated with GK1.5, > 90% of CD(+)(4) T cells were depleted; and in the mice treated with H-35, > 95% of CD(+)(8) T cells were depleted. Depletion of CD(+)(4) T cells could halt the recurrence of SK, reduce the severity of recurrent SK and impair the delayed type hypersensitivity. In contrast, depletion of CD(+)(8) T cells had no effect on the outcome of the disease. Histopathologic examination of the recurrent SK demonstrated that the stromal inflammation in CD(+)(8) T-cell depleted and control mice was characterized by extensive lymphocyte and polymorphonuclear leukocyte infiltration, but only occasional lymphocytes were observed in CD(+)(4) T-cell depleted mice. CONCLUSION: The results provide evidence that recurrent herpetic SK represents an immunopathologic process mediated by CD(+)(4) T cells. PMID- 11853579 TI - [An experimental study on treatment of limbal alkali burn by allograft transplantation with cultured stem cells on amniotic membrane]. AB - OBJECTIVE: To study the treatment of rabbit limbal alkali burn by cultured limbal stem cells growing on amniotic membrane for allograft transplantation. METHODS: After the primary culture of the rabbit corneal stem cells, they were cultured on amniotic membrane in DMEM/HamF12 medium for one week. Corneal stem cells and amniotic membrane were transplanted on the limbal and scleral area of the rabbit model with alkali burn. The corneal changes were observed by a slitlamp everyday, and the corneal pathological changes were examined. RESULTS: Cultured rabbit corneal stem cells continued to proliferate, differentiate and form multiple cell layers on amniotic membrane. After transplantation with cultured stem cells and amniotic membrane, the rabbit epithelium showed corneal phenotype and progressive decrease of vascularity and stromal infiltration in the limbal and peripheral zone. Pathological examination verified that the limbal and peripheral corneal epithelium was composed of multilayer cells, the neo-vascularization was reduced and stromal inflammatory cells were decreased. CONCLUSIONS: Allograft transplantation with cultured limbal stem cells can restore corneal epithelial cell composition, decrease neovascularization, maintain limbal cellular barrier function and provide better condition for keratoplasty later. PMID- 11853580 TI - [Experimental study on herpes simplex virus-1 functional gene latency in cornea]. AB - OBJECTIVE: To investigate the possible existence of herpes simplex virus-1 (HSV 1) functional gene latency in cornea. METHODS: A herpes simplex keratitis (HSK) model was established in New Zealand White (NZW) rabbit corneas, latently infected corneas were transplanted to one eye of each naive NZW rabbit. One week after the operation, the corneal buttons were removed and divided into three portions. One portion was processed for HSV-1 antigen detection, another was for HVS-1 latency associated transcript (LAT), thymidine kinase (TK) and DNA polymerase gene detections by polymerase chain reaction (PCR), and the last portion after tissue culture for 3 weeks, it was co-cultured with rabbit kidney cells (RK) for one week, then the expression of HSV-1 antigen in the RK cells and this portion of corneal button was detected. RESULT: In the 15 corneal samples negative for HSV-1 antigen, LAT gene was detected in 8 samples, TK gene in 10 and DNA polymerase gene in 6 samples. HSV-1 antigen was detected in 3 samples of RK cells and 6 cultured corneal samples. CONCLUSION: HSV-1 functional genes are retained in latently infected cornea. PMID- 11853581 TI - [Immunologic experimental studies on the alkali burn of cornea in rats]. AB - OBJECTIVE: To investigate the immune reaction of severe corneal alkali burns. METHODS: The models of severe corneal alkali burns were induced in rats. Immunohistochemistry was carried out on wholemounts of the cornea and iris at different time points after corneal burns to detect T-lymphocytes and their subpopulation, macrophages, dendritic cells and MHC class II -positive cells. RESULTS: In the early stage of the burns, there were small increasing number of T lymphocytes and MHC class II-positive cells in the cornea and iris. At the period of corneal melting and perforation, a massive influx of lymphocytes (CD(+)(3), CD(+)(4), CD(+)(8)), macrophages and dendritic cells was seen in the cornea adjacent to the limbus and iris. CD(+)(4) lymphocytes were much more than CD(+)(8) in number. T-cells and MHC class II-positive cells were observed not only at the limbus but also in the melting area and corneal center. Morphological changes of these cells were also noted. In the restoration stage, a number of positive cells were still found in the iris. CONCLUSION: Immunological mechanism is involved in the pathological changes of the damage induced by corneal alkali burns, especially in the pathogenesis of the corneal melting and perforation. PMID- 11853582 TI - [Effect on keratocyte-mediated collagen degradation by Pseudomonas aeruginosa]. AB - OBJECTIVE: To study the pathogenesis of cornea melting (ulceration) by pseudomona (P) aeruginosa for instruction of clinical treatment. METHODS: Type I collagen gels with or without suspended keratocytes were incubated for 24 hours under medium containing sterile P. aeruginosa culture broth. Native collagen fibrils were removed from the media by ultrafiltration. The ultrafiltrates were then hydrolyzed, and the amount of hydroxyproline was measured spectrophotometrically. The effect of a synthetic matrix metalloproteinase (MMP) inhibitor, Galardin, on collagen degradation was also examined. RESULTS: P. aeruginosa broth induced type I collagen gel degradation directly. In the presence of keratocytes, degradation by P. aeruginosa broth was enhanced. Galardin significantly reduced the amount of collagen degraded by P. aeruginosa culture broth, no matter keratocytes were present or not. CONCLUSION: P. aeruginosa culture broth directly degrades type I collagen and also increases keratocyte-mediated collagen degradation. The result is helpful to the clinical treatment of cornea melting caused by P. aeruginosa, and the mechanism should be further studied. PMID- 11853583 TI - [Multiple patterns of angle closure mechanisms in primary angle closure glaucoma in Chinese]. AB - OBJECTIVE: The mechanism of primary angle closure glaucoma (PACG) in Chinese is studied to establish a new classification system based on the etiology and mechanism of angle closure. METHODS: The anterior chamber angle's configuration and the anatomic structures related to the angle in 126 cases with PACG were observed with ultrasound biomicroscope (UBM), and the mechanisms of angle closure were analyzed by diagnostic treatment. RESULTS: Based on the configuration of the angle and anatomic structures related to the angle as well as the mechanisms of angle closure, PACG could be divided into following subtypes: pure pupillary block angle closure glaucoma [PPB, 48 cases (38.1%)]; pure non-pupillary angle closure glaucoma [PNP, 9 cases (7.1%)]; multiple mechanism angle closure glaucoma [MM, 69 cases (54.8%)]. PPB manifested that the relative position of pupillary margin was located anteriorly, the peripheral iris bombe was due to the pupillary block, and the angle closure was not related to the location and shape of the peripheral iris and ciliary body. PNP showed that the relative position of pupil margin was located posteriorly, the angle closure was not caused by the pupillary block, but caused by anterior located ciliary body and/or thick and anterior located peripheral iris. MM pattern was caused by both pupillary block and non pupilary block, and most of the cases manifested creeping angle closure. CONCLUSIONS: The angle closure mechanisms of PACG in Chinese are of multiple patterns. Based on the different mechanisms of angle closure in PACG, a new classification system should be established to improve the diagnosis and treatment of PACG. PMID- 11853584 TI - [The study of RDS gene mutation and clinical phenotype in a family with primary retinitis pigmentosa]. AB - OBJECTIVE: To investigate retinal degeneration slow (RDS) gene mutation in a Chinese family with primary retinitis pigmentosa (RP) and the association of the mutation with clinical phenotypes and to explore the pathogenesis of RP. METHODS: Blood DNA from 2 patients in the same family with RP and 2 normal persons was analyzed by molecular genetic methods. RDS gene mutation was screened out by polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) analysis. The mutant RDS gene fragment was cloned, then sequenced with an automatic DNA sequencer using a dideoxy chain termination protocol. The phenotype of the patients with the gene mutation were examined and determined by clinical ophthalmologic examinations. RESULTS: The PCR-RFLP analysis of the RDS gene in 2 patients with RP revealed codon 216 mutation of RDS gene. The mutation was heterozygous, and not found in 2 normal persons as controls. The alteration in the DNA sequence was identified as a heterozygous transversional change of C to T at the second nucleotide in codon 216 of RDS gene, resulting in the amino acid replacement of proline residue with leucine residue (Pro216Leu). The ocular finding of the patients with Pro216Leu mutation of RDS gene included severe visual loss and diffuse distribution of pigmentary changes with macular degeneration. CONCLUSIONS: The Pro216Leu mutation of RDS gene is found in Chinese patients with RP. The gene mutation is associated with the ocular phenotype, diffuse RP with macular degeneration. PMID- 11853585 TI - [A diagnostic analysis of imaging in ocular cysticercosis]. AB - OBJECTIVE: To discuss B-ultrasonography, CT and MRI appearances and image diagnostic value for ocular cysticercosis. METHOD: In the 29 cases with ocular cysticercosis, 21 cases were examined by B-ultrasonography, 19 cases by X-ray computed tomography (CT) and 6 cases by magnetic resonance imaging (MRI). RESULTS: At the living stage of the cysticercus, B-ultrasound might detect parasitic cyst and its movement in the eyeball and orbit, extraocular muscle irregular hyperplasia; CT might detect parasitic cyst in the orbit and the pathologic extraocular muscle; MRI might detect the parasitic cyst in the eyeball and the orbit and the pathologic extraocular muscle. At the non-living stage of the cysticercus, B-ultrasonography and CT might discover calcification focus in the eyeball and extraocular muscle. CONCLUSION: Image appearances of ocular cysticercosis depend on the parasitic site and living status of cysticercus in the eye region. For diagnosis, B-ultrasonography may first be selected, MRI secondly selected for living stage and CT secondly selected for non-living stage of the ocular cysticercosis. PMID- 11853586 TI - [A study on the effect of vitreous cavity length in primary ametropia]. AB - OBJECTIVE: To study the nature of primary ametropia and disclose the actions of ocular refractive components, especially the length of vitreous cavity, on primary ametropia. METHODS: The refractive state, corneal refractive power (CP), anterior ocular segment length (ASL), vitreous length (VL) and ocular axial length (AL) were measured for 1 336 eyes, including 523 hyperopic eyes, 199 emmetropic eyes and 614 myopic eyes. The relationships of these elements were studied. RESULTS: CP had no effect on the refractive state. There was no significant difference between CP of these three refractive groups (hyperopia vs. emmetropia and emmetropia vs. myopia) (P > 0.05). The test of normality showed that the distribution of CP was normal in each refractive state. Case control study of paired sample revealed that ASL did not change regularly with the change of refraction, while VL changed progressively with the change of refraction and there was significant difference between each pair of groups (F = 204.29, P < 0.01). The study of the relationship with matching t test between AL and VL showed that the change amount of AL and VL was nearly the same, that was, the growth of AL was about the same as the growth of VL (P > 0.05). Analysis of correlation and regression showed that these two elements were highly correlated, the coefficient of correlation being 0.879. CONCLUSION: VL determines AL and finally determines refractive state. VL is the most important element in determining primary ametropia while the anterior eye segment has no significant contribution. PMID- 11853587 TI - [Effects of synthetic interleukin-1 blockers on uveitis in rats]. AB - OBJECTIVE: To investigate the therapeutic action of synthetic interleukin-1 blockers on uveitis in rats. METHOD: Experiments were performed on 18 Spraque Dawley rats (36 eyes), and the retinal interleukin-1 beta (IL-1 beta) and tumor necrosis factor-alpha (TNF-alpha) were quantified by an enzyme-linked immunoabsorbent assay (ELISA) before and after the rats with uveitis treated by synthetic interleukin-1 blockers. RESULT: The retinal IL-1 beta and TNF-alpha in rats with uveitis treated by synthetic IL-1 blockers (CK-138, 139) were significantly decreased than those treated by contrasting design. CONCLUSION: Synthetic IL-1 blockers (CK-138, 139) are effective in treatment of IL-1 alpha induced uveitis in the rat. PMID- 11853588 TI - [Photoreceptor apoptosis and p53 gene expression in inherited retinal degeneration of RSC rats]. AB - OBJECTIVE: To investigate the gene regulation of photoreceptor apoptosis in inherited retinal degeneration. METHOD: The retinas of inherited retinal degeneration in animal models of RCS (Royal College of Surgeons) rats, ranging from 13(th) to 60(th) postnatal days in age, were studied by TdT-mediated biotin dUTP nickend labeling (TUNEL) for apoptosis, immunohistochemistry for p53 protein, in situ hybridization and reverse transcription-polymerase chain reaction (RT-PCR) for p53 mRNA. RESULTS: The photoreceptor of RCS rats underwent apoptosis from 25(th) postnatal day, and reached the highest level at 30 - 35(th) postnatal days. At the early stage of photoreceptor apoptosis, the level of p53 protein and mRNA in photoreceptors increased significantly. RT-PCR also showed that there was more production of p53 mRNA at 25 to 35(th) postnatal days in the retinas of RCS rats. CONCLUSION: P53 gene expression increases significantly at the early stage of photoreceptor apoptosis, indicating that p53 gene might play a role in the regulation of photoreceptor apoptosis in inherited retinal degeneration. PMID- 11853589 TI - [Anterior segment reconstruction and secondary posterior chamber intraocular lens implantation supported by fibromembrane]. AB - OBJECTIVE: To evaluate the methods and effects of anterior segment reconstruction and secondary posterior chamber intraocular lens (IOL) implantation supported by fibromembrane in pupillary area. METHODS: Anterior segment was reconstructed in 86 eyes with different anterior segment disorders. Anterior segment reconstruction includes: partial penetrating keratoplasty, loosing front adhesion, suturing detached peripheral iris, loosing rear adhesion, reshaping pupil, perforating fibromembrane in the pupillary area. The IOL was secondarily implanted into the posterior chamber, and the patients were followed up in 3 - 32 months. RESULTS: The surgery was successful in all patients. The postoperative visual acuities in 71 eyes were >or= 0.5 (82.6%), including >or= 0.9 (39.5%) in 34 eyes and 0.05). There were no significant differences in the mean SIA and uncorrected visual acuity between B and C groups throughout the whole study period (P > 0.05). CONCLUSIONS: SIAs in eyes of 3.2 mm incision group of phacoemulsification are less and more stable; a better and quicker rehabilitation of uncorrected visual acuity can be obtained in the early postoperative period. PMID- 11853591 TI - [The contribution of phacoemulsification to combined cataract and glaucoma surgery]. AB - OBJECTIVE: To study the safety and effectiveness of phacoemulsification (phaco) in trabeculectomy combined with cataract extraction (phacotrabeculectomy) and posterior chamber intraocular lens (IOL) implantation. METHODS: 68 eyes with glaucoma and cataract were randomly divided into two groups: the phacotrabeculectomy with IOL implantation and modern extracapsular cataract extraction (ECCE), trabeculectomy and IOL implantation. RESULTS: Postoperatively, in the phaco group, the intraocular pressure (IOP) decreased obviously at the early stage (P < 0.05), the number of cases necessary for the use of anti glaucoma drug was decreased, the mean kinds of the drug applied were markedly decreased (P < 0.05), the visual acuity improved quickly, significantly and steadily, and the complications were less than that in the ECCE group. CONCLUSIONS: The phaco's triple surgery is safe and effective. In cases with glaucoma and cataract, the surgery is indicated even when the cataract is not mature or premature. PMID- 11853592 TI - [Combined triple/multi-surgery of vitrectomy-lensectomy and posterior chamber intraocular lens implantation]. AB - OBJECTIVE: To study the feasibility and safety of the combined triple/multi surgery of vitrectomy-lensectomy and posterior chamber intraocular lens implantation. METHODS: Triple/multi-surgery was performed on 32 eyes of 32 cases, including 29 cases of serious eye injuries of traumatic cataract, lens luxation, vitreous hemorrhage (some cases with intraocular foreign body, iris damage or iridodialysis), 1 case with lens nucleus dislocated into the vitreous cavity during the senile cataract surgery and 2 cases with intraocular lens luxated into the vitreous cavity postoperatively. RESULTS: After follow up for 1 - 14 months (mean 5.6 months), 15 cases had the best corrected visual acuity (BCVA) >or= 0.5 (46.9%), 11 cases had BCVA of 0.1 - 0.4 (34.4%), and 6 cases had BCVA < 0.1 (18.8%). There was an average of 2.10 DC of astigmatism after surgery. Macular edema was found in 14 cases (43.8%). CONCLUSIONS: The results suggest that the triple/multi-surgery be safe and feasible. The major causes affecting the postoperative BCVA are astigmatism and macular edema. Proper selection of indication and skillful manipulation are the key points of the combined surgery. PMID- 11853593 TI - [Cause and management of dislocated nuclear fragments during phacoemulsification]. AB - OBJECTIVE: To investigate the cause and management of posteriorly dislocated lens nucleus in the vitreous cavity during phacoemulsification. METHODS: The authors performed a retrospective study of 18 patients with dislocation of nuclear fragment into vitreous cavity during phacoemulsification at Tong Ren Hospital from August 1994 to January 1999. RESULTS: 4 cases with dislocated nuclei floated in the anterior vitreous were removed by a lens loop from limbal extensive incision. 2 cases with their dislocated nuclei less than 1/4 of the normal size were under follow-up for 3 - 4 years, and no complications were observed. The final outcome in 1 case with a dislocated nucleus half of its normal size was ocular atrophy as a result of phacoanaphylactic endophthalmitis. 11 cases underwent pars plana vitrectomy. In the operation the nuclei were floated anteriorly by injection of perfluoro-1, 3-dimethylcyclohexane, then they were removed by a lens loop through the limbus and in 2 cases they were fragmented by ultrasound. The main post-operative complication was corneal edema. The final visual acuities generally were improved in varying degrees in 1 month to 4 years of follow-up. CONCLUSIONS: During phacoemulsification, the dislocation of a nucleus is liable to first occur in the peripheral sculpting stage, and secondly in central sculpting stage. A radial tear extending posteriorly from a discontinuous anterior capsulorrhexis is the major risk factor predisposing to posterior dislocation of the nucleus or nuclear fragment. During phacoemulsification, vitrectomy should be performed as soon as possible for a nucleus dislocated into middle or posterior vitreous cavity, and it is a safe and effective method for management of dislocated nucleus. PMID- 11853595 TI - [The ultrastructure of human and mouse cataractous lens epithelial cell apoptosis]. AB - OBJECTIVE: To observe the relation between human senile cataract, mouse congenital cataract and lens epithelial cell apoptosis. METHODS: Transmission electron microscope was used to observe the apoptotic morphology of the lens epithelial cells in 4 cases with senile cataract and in 2 mice with congenital cataract. The comparisons were made between the cataractous and normal transparent lens epithelial cells in human being and mice. RESULTS: The apoptotic cells were found in senile cataract and mouse congenital cataract lens epithelial cells, while in all control lens epithelial cells they were few. CONCLUSION: Lens epithelial cells apoptosis is related to senile cataract and mouse congenital cataract formation. PMID- 11853594 TI - [Vitrectomy for pseudophakic retinal detachment]. AB - OBJECTIVE: To study the method of vitrectomy for treatment of pseudophakic retinal detachment. METHODS: 32 pseudophakic retinal detachment eyes were treated with vitrectomy, primary vitrectomy in 11 eyes (34.4%), vitrectomy after conventional buckling failure in 21 eyes (65.6%), combined with silicone oil 10 eyes (31.3%), removal of intraocular lens 12 eyes (37.5%). All eyes had passed the 6 months to 5 years of follow-up examination. RESULTS: Intra-operative new retinal breaks were identified in 7 eyes (21.9%). The retinal total reattachment was in 25 eyes (78.1%); part reattachment was in 4 eyes (12.5%), and their visual acuity was improved than that before the operation. Due to severe proliferative vitreoretinopathy (PVR), failure occurred in 3 eyes. CONCLUSIONS: The main advantage of vitrectomy is the better intra-operative sight to the peripheral retinal breaks, removal of retinal traction and the low rate of PVR after vitrectomy. Vitrectomy for complicated type of pseudophakic retinal detachment can improve the success rate of the retinal detachment surgery and the visual acuity outcome. PMID- 11853596 TI - [Studies on intercellular communication of rabbit lens epithelial cells in vitro]. AB - OBJECTIVE: To investigate the intercellular communication in rabbit lens epithelial cells and effect of dexamethasone and heparin on it. METHODS: Rabbit lens epithelial cells were cultured and exposed to different concentrations of dexamethasone (10, 100, 300 mg/L), heparin (1 x 10(5), 2 x 10(5), 4 x 10(5) U/L) for 48 hours at 37 degrees C. The fluorescence redistribution after photobleaching (FRAP) was used for the analysis. The mean fluorescence recovery rate (MFRR, %/min) of the cells labelled with 6-carboxy fluorescein diacetate (CFDA) after photobleaching was measured with laser scanning cytometry ACAS Ultima. RESULTS: MFRR (%/min) of the cells after exposure to dexamethasone was 0.375 +/- 0.236 in the control group, 0.491 +/- 0.239 in the group of dexamethasone 10 mg/L, 0.996 +/- 0.447 in the group of dexamethasone 100 mg/L, 0.984 +/- 0.379 in the group of dexamethasone 300 mg/L, respectively (F = 26.07, P < 0.05). The MFRR after exposure of heparin was 0.375 +/- 0.236 in the control group, 0.694 +/- 0.491 in the group of heparin 1 x 10(5) U/L, 1.097 +/- 0.504 in the group of heparin 2 x 10(5) U/L, 1.082 +/- 0.501 in the group of heparin 4 x 10(5) U/L, respectively (F = 19.01, P < 0.05). CONCLUSION: It is discovered that there is intercellular communication in rabbit lens epithelial cells with FRAP analysis. Dexamethasone and heparin can enhance the intercellular communication of rabbit lens epithelial cells in dose dependent manner. PMID- 11853597 TI - [An experimental study on F-heparin surface modified IOLs implanted into the Rhesus monkeys' eyes]. AB - OBJECTIVE: To evaluate the biocompatibility of F-heparin surface modified IOLs. METHODS: 13 monkeys were divided into 3 groups, and implanted with different surface modified and non-modified IOLs into their eyes. All of the eyes were examined by slitlamp microscope and Schiotz tonometer at postoperative 15, 30, 60, 90 and 180 days. Postoperatively, at different periods the aqueous was aspirated from the anterior chamber to calculate cells. RESULTS: F-heparin surface modified IOLs induced milder inflammatory reaction and less dense posterior capsula opacification (PCO) than non-modified IOLs. CONCLUSION: F heparin surface modified IOL has a better biocompatibility. PMID- 11853598 TI - [Secondary glaucoma after penetrating keratoplasty]. AB - OBJECTIVE: To investigate the etiology and management of secondary glaucoma after penetrating keratoplasty (PKP). METHODS: Eighty cases (80 eyes) with secondary glaucoma occurring in 1,450 cases (1,500 eyes) of PKP, its causes, time of onset and the relationship with the primary disease were analyzed. According to different causes, they were treated with different therapies. RESULTS: The intraocular pressure (IOP) of 63 cases (63 eyes) became normal after drug treatment for 2 - 3 weeks. In 17 cases (17 eyes) who had received antiglaucomatous operations, 13 cases' (13 eyes) IOPs were normal and 4 cases' (4 eyes) still high. CONCLUSIONS: Accurate perioperative management can decrease the incidence of secondary glaucoma after PKP. The majority of secondary glaucoma can be controlled by different therapies according to different causes. PMID- 11853599 TI - [A clinical comparative study of interferon alpha-2b with mitomycin C applied in glaucoma filtering operation]. AB - OBJECTIVE: To compare the results of interferon alpha-2b (IFN alpha-2b) with mitomycin C (MMC) treatment in trabeculectomy. METHOD: 41 cases 68 eyes with late stage of primary open angle glaucoma (POAG) were randomly studied by treatment with IFN alpha-2b or MMC following trabeculectomy, each group consisting of 34 eyes. Of the cases, 27 were bilateral, one eye was designed to use IFN alpha-2b, and the other eye, apply MMC. All patients were ranged from 15 to 40 years old, and they all received the surgery the first time. The surgical procedure was similar in all eyes. The IFN alpha-2b treated eyes were subconjunctivally injected 5 x 10(5) IU at filtering bleb when the operation was finished immediately and on the postoperative day 3, 7, 10 and 14, respectively. The MMC receiving eyes were intraoperatively administered with 0.25 mg/ml via a sponge under the conjunctival flap for 5 minutes. The follow-up was ranged from 6 to 15 months. RESULTS: (1) The 12th month life-table success rate of functional bleb formation was (70.95 +/- 9.72)% in IFN alpha-2b-treated group and (77.01 +/- 10.51)% in MMC-received group, the difference being not statistically significant (u = 0.2165, P > 0.05). The eyes with IFN alpha-2b tended to form type II blebs were according to Kronfeld classification, whereas type I blebs were commonly seen in MMC-treated eyes (chi(2) = 6.261, P < 0.05). The percentages of intraocular pressure between 10 - 15 mmHg on postoperative month 3, 6, 9 and 12, respectively in IFN alpha-2b group were higher than that in MMC group. (2) The 12th month life-table complete and qualified success rates were (70.95 +/- 9.72)% and (94.59 +/- 5.26)% in IFN alpha-2b group, (65.15 +/- 10.51)% and (84.61 +/- 7.26)% in MMC group, respectively (u = 0.8174, 0.1011, P > 0.05). (3) Complications induced by IFN alpha-2b were rare, mainly involving transient corneal epithelial defect, whereas by MMC included thin-wall blebs, persistent hypotony and hypotonous maculopathy. The decrease of visual acuity was seen in 44.1% of the cases in MMC group, whereas only 17.6% occurred in IFN alpha-2b group (chi(2) = 5.217 4, P < 0.05). CONCLUSION: IFN alpha-2b has similar effect as MMC to reduce the risk of failure of glaucoma filtration surgery, but it has more advantages over because of its fewer complications. PMID- 11853600 TI - [Measurement of human retinal thickness at posterior pole with retinal thickness analyzer in normal and glaucomatous eyes]. AB - OBJECTIVE: To determine the diagnostic value of retinal thickness analyzer (RTA) in glaucoma. METHODS: RTA was used to measure the retinal thickness at the posterior pole in 77 normal persons (116 eyes) at various ages and 22 cases (35 eyes) with glaucoma. The obtained images were processed by a computer to yield the retinal thickness value and the thickness map at this location, and the data were analyzed with SAS software package. RESULTS: The mean retina thickness of normal subjects was (171.83 +/- 18.50) microm with no significant difference among the various age groups (F = 2.68, P > 0.05), and very significant between the sexes; the thickest retina was at the location between macula and optic nerve head, and the macular nasal retina is 11.7% thicker than its temporal side. The upper and lower retinas of the fovea were basically symmetrical. The retinal thickness in glaucoma patients showed diffuse or local thinning, and there was a very significant difference between the normal group and the glaucomatous group in retina thickness (F = 11.98, P = 0.0001). The sensitivity rate for detecting glaucoma with RTA was up to 80.8%, and specificity to 78.5%. CONCLUSIONS: The thickness map for the retina at posterior pole across the macula in normal subjects is "horse shoe" shaped, that is well matched with the topography of retinal ganglion cell and the retinal nerve fibers in human retina. Diffuse or local thinning of retinal thickness in glaucomatous patients, which is in agreement with the corresponding visual field defect, and the sensitivity of RTA is higher than that of perimeter. The RTA examination is easy to operate, has high reproducibility, no invasion, therefore it is an ideal method to measure the human retinal thickness in vivo. PMID- 11853601 TI - [Poly-static quantitative perimetry for detection of open angle glaucoma]. AB - OBJECTIVE: To evaluate the sensitivity of poly-static quantitative perimetry for the detection of visual field defect in open angle glaucoma. METHODS: 48 patients (95 eyes) of open angle glaucoma were examined with poly-static quantitative perimetry [Friedmann visual field analyzer (FVFA)] and dynamic perimetry (tangent screen) respectively. RESULTS: 26 eyes showed small defect of visual field on tangent screen with 2/1,000 visual size, and 23 eyes (88%) showed the same defects checked with FVFA; visual field defects of early stage glaucoma were found in 26 eyes checked with FVFA, but only 16 eyes showed the same defects on tangent screen with 1/1,000 visual size; the visual field defects of glaucoma on tangent screen in 43 eyes were found larger when they were checked with FVFA. CONCLUSION: Comparing with dynamic perimetry, poly-static quantitative perimetry is more sensitive to detect visual field defect of early stage glaucoma. PMID- 11853602 TI - [The effect of artificial tears on corneal surface regularity in dry eye]. AB - OBJECTIVE: To evaluate the regularity of corneal surface and the effect of artificial tears on the corneal surface regularity in dry eye. METHODS: The surface regularity index (SRI), surface asymmetry index (SAI), the indices of TMS 1 corneal topography system, and potential visual acuity (PVA) were used to investigate the corneal regularity and corneal vision in 33 normal subjects (64 eyes) and 22 dry eye patients (42 eyes) before and after instillation of artificial tears. RESULTS: The SRI and SAI were significantly elevated and the PVA was remarkably decreased in patients with dry eye compared to that of normal subjects (P < 0.001). The average amount of astigmatism was also significantly increased in dry eye compared to that in normal eyes (P = 0.02). In the patients with dry eye, the SRI and SAI were positively correlated with corneal fluorescein staining scores. The SRI, SAI and astigmatism were significantly reduced, the PVA was improved and the irregular topographic pattern was markedly decreased in the dry eyes after the usage of artificial tears. CONCLUSION: The dry eye has an irregular corneal surface and the indices of TMS-1, SRI and SAI can be used to objectively diagnose this disorder, evaluate the severity of the dry eye and the effect of artificial tears on it. Artificial tears can smooth the corneal surface and improve the visual acuity in patients with dry eye. PMID- 11853603 TI - [Analysis of influence of axial length and keratometric value on the curative effects of excimer laser photorefractive keratectomy for myopia]. AB - OBJECTIVE: To evaluate the influence of axial length (AL) and keratometric value (KV) on the curative effects of excimer laser photorefractive keratectomy (PRK) for myopia. METHOD: A Keracor-17 excimer laser was used to treat 80 patients (121 eyes) with myopia. The cases were divided into three groups according to the AL and KV: The first group AL > 26 mm, KV 45 D, 36 eyes; the third group AL 0.05). The cases with uncorrected visual acuity >or= 0.5 had significant difference between the first and the second group and between the first and the third group (chi(2) = 6.30, P < 0.05). Multiple linear regression and correlative analysis showed that the increase in ocular axis was an unfavorable factor to visual acuity, while the increase in keratometric value was a favorable factor to visual acuity. CONCLUSION: PRK for treatment of myopia is an effective and safe refractive surgery. When the myopic degrees are identical, the postoperative outcome of myopia with increase in keratometric value as the main pathogeny is better than the myopia with increase in axial length as the main pathogeny. PMID- 11853604 TI - [Spectrum of mycotic keratitis in China]. AB - OBJECTIVE: To investigate the spectrum of mycotic keratitis in China. METHODS: We scraped and evaluated 615 mycotic keratitis in microbiology laboratories of Zhengzhou (central China), Shijiazhuang (north China) and Guangzhou (south China) in the twenty-two years between Jan. 1, 1975 and Jun. 30, 1997. RESULTS: Organisms of 18 genuses and 615 species isolated from corneal ulcers were found. Fusarium species were isolated the most frequently (65.0% in Zhengzhou, 33.3% in Shijiazhuang and 39.2% in Guangzhou), followed by Aspergillus (20.5% in Zhengzhou, 21.7% in Shijiazhuang, 30.7% in Guangzhou), Penicillium (3.6% in Zhengzhou, 11.6% in Shijiazhuang) and Curvularis (13.1% in Guangzhou). Between Jan. 1, 1975 and Dec. 31, 1986, Aspergillus species were most commonly isolated in Guangzhou (48.5%), followed by Fusarium (27.7%) and Penicillium (14.27%). CONCLUSIONS: Just as different regions of the world are characterized by specific endemic infections, the predominating organisms in corneal fungal infections also vary throughout the world. These regional differences in causative organisms in mycotic keratitis are clinically important because they influence the initial and more definite therapy. This study shows that Fusarium species are the most common organisms of mycotic keratitis in China. PMID- 11853605 TI - [A descriptive epidemiological investigation of anophthalmos and microphthalmos in China during 1988 - 1992]. AB - OBJECTIVE: To investigate the descriptive epidemiological characteristics of cases with anophthalmos and microphthalmos in China. METHOD: According to the hospital-bases monitoring method, the birth defects monitoring program was undertaken in 443 - 588 hospitals from 30 provinces, cities and autonomous regions in China. Data of the new born babies including intra-uterine death and stillbirth from 28 weeks of gestation to a period of 7 days after birth were collected between 1988 and 1992. RESULTS: There were 3,246,408 babies monitored, among which 382 cases of anophthalmos and microphthalmos were found. The average prevalence rate was 1.18/10,000 in China. The decreasing tendency of prevalence rate was shown during the period (chi(2) = 7.381, P < 0.01). The average prevalence rate in the rural area was significantly higher than that in the urban area, and the female cases were higher than that of male cases. The prevalence rates among various regions varied from 0.21 to 2.29/10,000 with the highest in Gansu province and lowest in Tianjin city. 87.7% of the cases with anophthalmos and microphthalmos were associated with other congenital malformations (multiple malformations). The associated malformations were mostly facial (including ear, neck, etc.) followed by skeletomuscular system and central nervous system. 8.1% of cases were diagnosed as syndromes, among which trisomy 21 was the most common condition. CONCLUSIONS: There were large variations in the prevalence rates of anophthalmos and microphthalmos in different parts in China. Careful analysis, particularly the chromosome analysis, should be performed to correctly diagnose the cases, especially for those with multiple malformations. PMID- 11853606 TI - [The histogenesis, clinical features and histopathological analysis on 52 cases of ciliary body neoplasms]. AB - OBJECTIVE: To analyze the histogenesis, classification, incidence, clinical feature, the key point of differential diagnosis and histopathologic characteristics of ciliary body neoplasms to provide the basis for clinical diagnosis and treatment. METHOD: 52 specimens of ciliary body neoplasms on file were analyzed and studied for the characteristics of clinical presentation and histopathology. RESULTS: Of the 52 specimens, 38 were located in the ciliary body (73.1%), and 14 lesions involved the iris (26.9%). According to the results of histogenetic classification, the tumors of ciliary sensory epithelium were ranked the first place (20 cases, 38.5%) while malignant melanoma, the second one (14 cases, 26.9%) followed by melanocytoma (9 cases, 17.3%) and granulomatous lesions (5 cases, 9.6%). CONCLUSION: The clinical features and histopathologic characteristics of ciliary body neoplasms are systemically and deeply recognized, that may help clinical ophthalmologists to make correct diagnosis and proper treatment. PMID- 11853607 TI - [Apoptosis and related gene expression in lacrimal gland of cases with Sjogren's syndrome]. AB - OBJECTIVE: To assess the roles of apoptosis and expression of the related genes in lacrimal glandular destruction of patients with Sjogren's syndrome (SS). METHODS: Small pieces of palpebral lobes of lacrimal glands were obtained from 12 patients (10 with primary Sjogren's syndrome, 2 with secondary Sjogren's syndrome) and 7 normal controls. They had been investigated by the in situ end labeling and immunohistochemical staining to detect the apoptotic cells and the expression of Fas, FasL, bcl-2 and Bax. RESULTS: The number of epithelial apoptotic cells in lacrimal glands of patients with SS was significantly higher than that in normal glands, and SS epithelial cells showed increased expression of Fas, FasL and Bax. There was significant positive correlation between the number of the apoptotic cells and that of the cells expressing Fas, FasL and Bax, respectively, and there was significant negative correlation between the number of the apoptotic cells and that of the cells expressing bcl-2. CONCLUSIONS: The apoptosis of the epithelial cells in the lacrimal gland may be one of the mechanisms leading to the glandular destruction found in SS. In SS lacrimal glands, the expression of Fas, FasL and Bax may promote apoptosis, and the expression of bcl-2 may inhibit apoptosis. PMID- 11853608 TI - [A study on the expression of p16 protein and bcl-2 protein in cutaneous eyelid tumors]. AB - OBJECTIVE: To investigate the relationship between p16 (tumor suppressor) and bcl 2 (apoptosis inhibitor) gene expression and the incidence and development of eyelid malignant tumors. METHODS: Streptavidin-biotin-peroxidase complex immunohistochemical method was used to study the expression of p16 gene and bcl-2 gene in 96 cases with eyelid malignant tumors. RESULTS: In the 96 cases, including 40 cases of basal cell carcinoma (BCC), 33 squamous and 23 sebaceous carcinomas, their p16 protein positive (nuclear staining) rates were 70.0%, 54.6% and 56.5% respectively. The p16 positive rate was negatively correlated to the degree of tumor histological differentiation, and the rate difference between the high differentiated carcinoma and the low differentiated one was significant (P < 0.05). bcl-2 protein positive expression was detected in the cytoplasm. All of the 40 BCCs were bcl-2 positive and nearly all the tumor cells showed positive cytoplasmic expression, while in the 33 specimens of squamous cell carcinoma, only did one show positive focal reaction and the staining in other 32 cases was relatively faint. None of the 23 sebaceous carcinoma expressed bcl-2. CONCLUSIONS: The expression of p16 protein is related to the occurrence and degree of differentiation of eyelid malignant tumors. The over-expression of bcl 2 protein suggests that suppression of apoptosis might play a role in the tumorigenesis of BCC. PMID- 11853609 TI - [An observation of antiproliferative effect of germanium-132 on cultured pterygium fibroblasts]. AB - OBJECTIVE: To detect the antiproliferation of carboxyethyl germanium sesquioxide (Ge-132) on fibroblasts of pterygium in vitro and try to find a potentially effective agent for treatment of primary pterygium and prevention of its postoperative recurrence. METHODS: Primary culture and subculture of pterygium fibroblasts were established in vitro. Different concentrations of Ge-132 (39 - 5,000 mg/L) or mitomycin-C (3.13 - 4.00 mg/L, the control) were added to the fibroblast culture of the third or forth passage respectively. The inhibitory effect was determined by MTT (tetrazolium bromide) method. The influence of addition of Ge-132 on the growth curve of fibroblasts was observed, and the changing expression of proliferating cell nuclear antigen (PCNA) in fibroblasts was studied by immunohistochemical method. RESULTS: The addition of Ge-132 in the culture caused significant inhibition of the fibroblast proliferation in dose dependent manner (625 - 50,000 mg/L) without cytotoxicity (IC(50) = 3,000 mg/L), the marked descent of growth curve and suppression of the expression of PCNA in cultured cells (P < 0.01). CONCLUSION: Ge-132 can inhibit the proliferation of pterygium fibroblast in vitro significantly. PMID- 11853611 TI - [MRI features of confusable tumors of orbital apex]. AB - OBJECTIVE: To analyze magnetic resonance imaging (MRI) features of confusable tumors including cavernous hemangioma, cavernous lymphangioma and neurilemmoma located at orbital apex, which could provide evidences for diagnosis and surgical treatment. METHODS: MRI was performed on 16 cases with tumors of orbital apex confirmed by pathology, and their MRI findings were analyzed. RESULTS: The time taken to attain complete enhancement after contrast on MRI was significantly different in the three types of tumor (P < 0.01) Their enhancement patterns were related to their pathologic findings. CONCLUSIONS: The different enhancement patterns of various tumors of orbital apex are characteristic for diagnosis and differential diagnosis. PMID- 11853610 TI - [Point mutations of N-ras oncogene and abnormal expression of rasp21, p53 proteins in orbital rhabdomyosarcoma]. AB - OBJECTIVE: The point mutations of N-ras oncogene and the abnormal expression of rasp21 and p53 proteins in orbital rhabdomyosarcoma (RMS) were analyzed in order to explore the role of oncogene mutations in the pathogenesis of orbital RMS and its relation to patients' prognosis. METHOD: 21 human orbital RMS tissues were analyzed by dot-blot hybridization and immunohistochemistry using synthetic specific oligonucleotide probes and mutant rasp21, p53 monoclonal antibodies. RESULTS: The second base mutation of codon 12 of the N-ras oncogene was found in 4 patients. The third base mutation of codon 61 was found in 5 patients. The mutant rate was 33.3%. By immunohistochemistry analysis, the overexpression of rasp21 and p53 protein level was found in 42.9% and 76.2% respectively. The prognosis of patients with overexpression of ras p21, p53 was worse than that of normal expression (P < 0.01). CONCLUSION: The mutants of ras and p53 oncogene play important roles in the pathogenesis of orbital RMS, and the abnormal expression of their protein products is related to patients' prognosis. PMID- 11853612 TI - [An experimental research of taurine on H2O2-induced bovine lens epithelial cell apoptosis]. AB - OBJECTIVE: To study the inhibitory role of taurine on H(2)O(2)-induced bovine lens epithelial cell apoptosis. METHODS: By terminal deoxyribonucleotide transferase-mediated dUTP nick end labeling (TUNEL) detection and bovine lens organ culture, we examined the affect of taurine on the number of H(2)O(2) induced bovine lens epithelial cells with apoptosis, the related comparisons and statistic analyses were carried out. RESULTS: (1) Lens epithelial cell apoptosis began before lens opaqueness (lens became opaque after 6 hours of incubation, while the lens epithelial cell apoptosis was detected after 3 hours of incubation); (2) Taurine could apparently inhibit H(2)O(2)-induced bovine lens epithelial cell apoptosis. CONCLUSION: Taurine has an inhibitory role on H(2)O(2) induced bovine lens epithelial cell apoptosis and can delay and ameliorate the occurrence and development of cataract. PMID- 11853613 TI - [A study on correlation between the structural parameter of optic disc and visual field in primary open-angle glaucoma]. AB - OBJECTIVE: To study the consistency between the optic disc neural rim depth (NRD) and visual field defect in patients with primary open angle glaucoma (POAG). METHODS: A system of computerized image analysis was used to measure the relative NRDs of temporal-superior and temporal-inferior area of the optic disc in patients with primary open angle glaucoma. The consistency between the area with relatively larger NRD and the location of severer visual field defect was investigated. RESULTS: In cases with early and middle stage POAG, the value of NRD corresponding to the severer location of the visual field defect was significantly larger than that corresponding to the location without or with mild such defect. Among 54 eyes, there were 46 eyes whose location with larger NRD value was consistent with the position of severer visual field defect, the rate of consistency being 85.2%. CONCLUSION: In cases with early and middle stage POAG, the increase of NRD value at temporal-superior and temporal-inferior area is asymmetrical, and the consistency between the area with larger NRD value and the corresponding position of severer visual field defect is relatively good. NRD is helpful to the early diagnosis of POAG. PMID- 11853614 TI - [The detection of disparity evoked potentials in anisometropes]. AB - OBJECTIVE: To investigate the affection of anisometropia on stereopsis and its mechanism. METHODS: A new set of static random-dot stereograms was utilized as a stimulus to elicit evoked potentials in 20 anisometropes and 40 normal subjects. RESULTS: Under the stimulus of different degrees of disparity in anisometropes, the P(250) waves related to stereopsis could be recorded. However, the percentage of their P(250) wave amplitude over the plane figure was significantly lower than that of normal persons (P < 0.05). The P(250) wave mean amplitude of severe anisometropes (aniseikonia >or= 5%) in fine disparity (14' and 23') was lower than that in mild anisometropes (aniseikonia < 5%, P < 0.05). CONCLUSIONS: Anisometropia may affect and disturb the stereopsis. The degree of disturbance is related to the degree of anisometropia. The higher the degree of anisometropia, the lower the amplitude of the P(250) potential, and the main defect is at the part of fine disparity. PMID- 11853615 TI - [A clinical investigation on cataract surgery with 2.8 mm incision]. AB - OBJECTIVE: The results of phacoemulsification which was performed through a 2.8 mm clear cornea tunnel incision and foldable intraocular lens (IOL) implantation were analyzed. METHODS: On 126 eyes with cataract of 105 cases, phacoemulsification with foldable IOL implantation was randomly performed. The results of postoperative vision, refraction, corneal edema, anterior chamber flare and the rate of corneal endothelial cell loss were retrospectively summarized. RESULTS: The uncorrected visual acuities of postoperative 1 day, 3 days, 1 week, 2 - 3 months, >or= 0.5 were in 102 eyes (81.0%), 108 eyes (85.7%), 112 eyes (88.9%) and 112 eyes (88.9%), respectively. The mean diopters of astigmatism at postoperative 3 days, 1 week, 1 month and 3 months were (1.09 +/- 0.51) D, (1.02 +/- 0.47) D, (0.88 +/- 0.52) D and (0.81 +/- 0.62) D, respectively. The mean diopters of astigmatism at postoperative 1 week, 1 month and 3 months compared with that before the surgery had no significant difference (P > 0.05). The comparison between the preoperative astigmatism and that at postoperative 3 months showed that only about 0.08D of change existed. The postoperative corneal edema and anterior chamber flare were mild, the anterior chamber configuration remained well, and the rate of endothelial cell loss was 11.9%. CONCLUSIONS: The surgical incision is small and easy to perform. Postoperatively, the reaction is mild, the change of astigmatism is small in degree, the best visual acuity can be obtained at the early stage and the refractive status is stable, thus the duration for restoration is greatly shortened. PMID- 11853617 TI - [The affect of Erigeron Breviscapus (Vant.) Hand-Mazz on axoplasmic transport of optic nerve in rats with experimentally elevated intraocular pressure]. AB - OBJECTIVE: To investigate whether Erigeron Breviscapus (Vant.) Hand-Mazz (EBHM) can improve the optic nerve axoplasmic transport in rats with experimentally elevated intraocular pressure (IOP). METHODS: Thirty healthy SD rats were used for the study, acute elevated IOP model in the right eye was built, then they were divided into three groups randomly: Group A (0 day group) included six rats for retinal ganglion cell (RGC) counting via left superior colliculus retrograde horse radish perokidase labeling; Group B, twelve rats divided into EBHM treatment group and control group (6 rats in each subgroup) for RGC counting via left superior colliculus retrograde labeling after twenty days, and Group C included twelve rats submitted the same treatment and procedure as group B after 40 days. RESULTS: After 0 day of acute elevated IOP, no labeled RGCs were observed. After twenty days of acute elevated IOP, in the control and EBHM subgroups the density of labeled RGCs were (423 +/- 220)/mm(2) and (749 +/- 294)/mm(2) respectively, the difference between two subgroups showed statistical significance (P < 0.01). After 40 days of acute elevated IOP, the density of RGCs in the control and EBHM subgroups in group C were (610 +/- 315)/mm(2) and (1,048 +/- 393)/mm(2) respectively, the difference between the two subgroups being statistically significant (P < 0.01). CONCLUSION: After 20 days and 40 days of acute elevation of IOP, the density of RGCs is obviously higher in EBHM group than that in the control group. It is revealed that EBHM can improve the optic nerve axoplasmic transportation blocked by acute elevation of IOP in rats. PMID- 11853616 TI - [A comparison between latanoprost and timolol in treatment of patients with primary open-angle glaucoma and ocular hypertension]. AB - OBJECTIVE: To evaluate the therapeutic value of latanoprost on glaucoma. METHODS: In an open-label fashion, multicenter, randomized control clinical trial, the efficacy and adverse drug reactions of topical application of 0.005% latanoprost once daily were compared with that of 0.5% timolol twice daily for 12 weeks in patients with open-angle glaucoma or ocular hypertension. RESULTS: The study included 128 patients (63 patients in latanoprost group and 65 patients in timolol group) and 117 patients remained at the end of the study (60 cases in latanoprost group and 57 cases in timolol group). Comparing 12 weeks with baseline diurnal intraocular pressure (IOP), the IOP reduction (mean +/- standard deviation) in latanoprost group was (7.5 +/- 0.3) mm Hg (1 mm Hg = 0.133 kPa) (32%, t = 22.73, P < 0.0001) greater than the reduction in timolol group (6.1 +/- 0.3) mm Hg (26%, t = 17.94, P < 0.0001), the difference between the two groups being significant (F = 9.54, P = 0.0026). Two patients treated with timolol and none treated with latanoprost were withdrawn from the study because of inadequate IOP control; 3 patients with latanoprost had foreign body sensation. In latanoprost group, there was one patient whose eyelashes became darker and longer at the last visit (the 12th week). No ocular and systemic adverse events related to the two drugs were found. CONCLUSION: It is demonstrated that 0.005% latanoprost topically applied once daily is well tolerated and more effective in reducing IOP than 0.5% timolol topically applied twice daily. Thus, latanoprost has the potential to be a new first-line antiglaucoma drug. PMID- 11853618 TI - [Suppressing effect of isoforskolin and forskolin on ocular hypertension in rabbits]. AB - OBJECTIVE: To examine the effects of isoforskolin (isoF) and forskolin (FSK) suspensions administered topically on eyes with ocular hypertension (OHT) and observe the local irritation of the drugs. METHODS: The intraocular pressure (IOP) was measured with pneumotonometer in screened spontaneous or water-loading induced OHT rabbits for evaluation of drug effects, and the irritation indices were recorded as well. RESULTS: In screened OHT rabbits, isoF significantly lowered IOP from 0.5 to at least 4.5 h after the administration (P < 0.05, paired t-test vs. contralateral control eyes), with the maximum decreases of 0.25% and 1% isoF reaching 8.1 mm Hg and 7.0 mm Hg (1 mm Hg = 0.133 kPa) at about 1.5 h later, respectively. In water-loaded OHT rabbits, both 0.06%, 0.1%, 0.25%, 1% isoF and 1% FSK concentration-dependently suppressed the increments of IOP (P < 0.05 with the exception of 0.06% isoF) induced by water-loading, and the peak effects of the optimal concentration of 0.25% isoF and 1% FSK were 7.1 and 6.9 mm Hg (difference of IOP between bilateral eyes) respectively. The results also indicated that 0.25% isoF or FSK suspension had basically no irritation, but 1% induced slight irritating responses during the experiment. CONCLUSION: Both isoF and FSK had suppressing effects on OHT in rabbits with the optimal concentration being 0.25% and 1% respectively; at concentration of 0.25%, topically both drugs had basically no irritation, while 1% of the drugs may induce mild irritating responses. PMID- 11853619 TI - [Pressure influence on mRNA expression and protein synthesis of inducible nitric oxide synthetase in bovine trabecular meshwork cell]. AB - OBJECTIVE: To investigate the inducible nitric oxide synthetase (iNOS) mRNA expression and protein synthesis in bovine trabecular meshwork cell and discuss the possible effects of nitric oxide (NO) in the development of glaucoma. METHODS: 20 mm Hg, 40 mm Hg, 60 mm Hg and 80 mm Hg pressure were respectively added on cultured trabecular cells of new born bovine. No pressure group was set as the control. The changes of iNOS mRNA and protein in trabecular meshwork cells under different pressures were demonstrated qualitatively and quantitatively by in situ hybridization and nicotinamide-adenine dinucleotide phosphate diaphorase (NADPH-d) histochemical assay. RESULTS: The expression of iNOS mRNA and its protein synthesis became higher and higher as the pressure was increased. There was weak iNOS mRNA expression in the control and 20 mm Hg groups, and no significant difference between them. 40 mm Hg and 60 mm Hg groups had a statistical difference from the control group. And 80 mm Hg group had a very significant difference from the control group. There was no difference between 40 mm Hg and 60 mm Hg groups, but they had significant difference from 80 mm Hg group. NADPH-d histochemical assay showed almost the same results except that there was significant difference between 40 mm Hg and 60 mm Hg groups. CONCLUSIONS: Pressure can evoke the expression of iNOS mRNA, and the nitric oxide thus produced can be one of the causes of trabecular meshwork destruction, that may induce or aggravate glaucoma. PMID- 11853620 TI - [The effect of dactinomycin on retinal injury induced by argon laser in mouse]. AB - OBJECTIVE: To determine that the administration of RNA synthesis inhibitor (dactinomycin) prevents retinal photoreceptor cell injury induced by green wavelength of argon laser in SD mice. METHODS: At various time (0, 8, 20 h) after the exposure to laser, eyes of the mice were injected intraocularly with dactinomycin in experimental eyes, and saline in control ones, and the concentration of the drug were 10 g/L, 5 g/L, 2.5 g/L. The eyes were taken at 24 h after the exposure, the slices of the retina were stained with methyl green pyronin (MG-P), and pyknotic (violet) as well as surviving cells (green) were counted in the laser burns. RESULTS: Intraocular administration of dactinomycin diminished the degeneration induced by argon laser, there were marked differences between the experimental group and the control group, dactinomycin showed to be more effective if administered immediately after exposure and 8 h later, after this time the higher concentration of the drug could not result in higher effect. CONCLUSION: Dactinomycin can inhibit the occurrence of the laser injury, but the time selected for the administration is critical for its efficacy. PMID- 11853621 TI - [Combined phacoemulsification, foldable intraocular lens implantation and trabeculectomy for cataract patients with glaucoma]. AB - OBJECTIVE: To evaluate the therapeutic effects and safety of the combined phacoemulsification, foldable intraocular lens implantation and trabeculectomy (triple procedure) and compare the outcomes of the operation with two different small incisions, conventional scleral flap and scleral tunnel flap. METHODS: Triple procedure was performed through a 3.5 mm incision by means of conventional scleral flap or scleral tunnel flap on 44 eyes of 42 patients with cataract and glaucoma. RESULTS: After follow-up for 3 -- 6 months, the mean intraocular pressure (IOP) was (14.33 +/- 3.68) mm Hg (1 mm Hg = 0.133 kPa) with a mean pressure reduction of 10.35 mm Hg from that before the operation (P < 0.001). Postoperatively, the visual acuities were >or= 0.5 in 33 eyes (75.0%) at 1 week and in 37 eyes (84.1%) during 3 - 6 months. The mean postoperative astigmatism was (1.29 +/- 0.93) D at 1 week and only 0.17D more than the mean preoperative one (P > 0.05). There was also no statistical difference between the mean preoperative astigmatism and postoperative astigmatism at 3 - 6 months, which was (1.12 +/- 0.73) D. The filtering blebs of type I and II (functional) by Kronfeld's classification presented in 79.5% of the eyes and type III (non functional) in 20.5% of the eyes. There were no statistical differences for the postoperative outcomes of IOP, visual acuity, astigmatism and filtering bleb between two different procedure groups (P > 0.05). CONCLUSIONS: The triple procedure is an effective, quick, repeatable and safe procedure for cataract patients with glaucoma. There are no differences at the surgical outcomes of two different incisions by means of the conventional scleral flap vs scleral tunnel flap. PMID- 11853622 TI - [Cataract extraction with rotated-chipping phacoemulsification in the capsule]. AB - OBJECTIVE: To observe the clinical effects of rotated-chipping phacoemulsification in the capsule. METHODS: The operation of continuous circular capsulectomy, nuclear hydrodissection, cataract extraction with rotated-chipping phacoemulsification in capsule and intraocular lens implantation was performed on 215 eyes. RESULTS: Rotated-chipping technique was successfully completed in 215 eyes. In the operation 2 to 4 circles were rotated with the ultrasonic energy 19.3% for 12 seconds on average. The rate of corneal endothelium loss was 7.5%. CONCLUSION: The technique of rotated-chipping phacoemulsification in capsule is safe and rapid which can be used for cataracts with nuclei of various degrees in sclerosis. PMID- 11853623 TI - [The nuclear hardness and associated factors of age-related cataract]. AB - OBJECTIVE: To determine the nuclear hardness and associated factors of age related cataract. METHODS: Forty-seven eyes of 47 patients with age-related cataract aged > 50 years were studied. The opacity of lenses was assessed and classified by the LOCS II pre-operatively. The lens nucleus obtained during extracapsular cataract extraction (ECCE) was measured. By means of a fine conical probe and dynamometer, the resistance to penetration of different lens layers was transferred to an electric signal and recorded by a function recorder. The hardness to the penetration of different lens layers was calculated. Multivariate regression analysis was applied to the parameters including age, color of the nucleus, duration of symptoms and other factors. RESULTS: The mean force required to penetrate the lens was (0.51 +/- 0.29) Newtons (N) for grade I, (1.41 +/- 0.35) N for grade II and (1.95 +/- 0.42) N for grade III; it increased to (2.96 +/- 0.62) N for grade IV. The hardest part of the lens was located at the nucleus center. Multivariate analysis of the data showed that 85% of the variation in hardness could be explained in terms of color and opacity. The patients with myopia and diabetics had an increased incidence of grading IV nuclear sclerosis. Regression analysis revealed a positive correlation between nucleus color and weight (r = 0.4565), diameter (r = 0.5263), thickness (r = 0.5552) and hardness (r = 0.8969), P < 0.001. There was a weak positive correlation between age and the weight of nucleus (r = 0.3239, P < 0.05). CONCLUSIONS: The results indicate that the color can be used more reliably to predict physical characteristics of lens nucleus, the preoperative knowledge of which would help the surgeon in planning small-incision surgery including phacoemulsification. PMID- 11853624 TI - [The age-related effect of basic fibroblast growth factor on the proliferation of lens epithelial cells]. AB - OBJECTIVE: To detect the age-related effect of basic fibroblast growth factor (bFGF) on the proliferation of human lens epithelial cells (HLECs) in patients with cataract and discuss the mechanism. METHODS: Human lens anterior capsules with attached HLECs were cultured in vitro, treated with 10 microg/L bFGF for 48hr, and then the positive area ratio of proliferating cell nuclear antigen (PCNA) in immunohistochemistry was estimated. Immunohistochemical analysis of bFGF receptor was also carried out. RESULTS: The bFGF could enhance the HLEC proliferation in patients with cataract at various ages. The enhancement of the proliferation and the expression of bFGF receptor protein were negatively correlated to the age of the patients with cataract (P < 0.01). CONCLUSIONS: The basic fibroblast growth factor might play an important role in the formation of posterior capsular opacity (PCO) after cataract surgery. With the decrease in the patient's age, the enhancement of HLEC proliferation is increased, indicating that the fact is related to the high incidence of PCO in children. PMID- 11853625 TI - [The early biochemical changes of cataractous lenses of rats cultured in vitro]. AB - OBJECTIVE: To study the mechanism of cataractogenesis. METHODS: Tissue culture was used to study the cataractous lenses of rats induced by sodium selenite or galactose. At the early stage, the content of nonprotein sulfhydryl (NP-SH), protein sulfhydryl (P-SH), non-soluble disulfide bond, malonaldehyde (MDA) and the activities of glutathione peroxidase (GSH-PX), glutathione reductase (GSSG-R) and glutathione-S-transferase (GSH-S) were measured in the cataractous lenses, and they were compared with that of the normal lenses. RESULTS: The two kinds of material could all induce cataract in rats, sodium selenite being more potent. In the early period of culture (lenses were transparent), NP-SH and P-SH were decreased, while disulfide and MDA were increased, the activity of GSH-PX rose obviously, that of GSH-S also had a tendency of rise, however, the activity of GSSG-R had no obvious changes. CONCLUSIONS: Rat lens opacity may occur after the lens is cultured in vitro with the addition of sodium selenite or galactose, and biochemical changes may develop in the lens at the early period of culture (lenses are transparent). PMID- 11853626 TI - [Adherence of bovine lens epithelial cells to the surface of intraocular lens made of different materials]. AB - OBJECTIVE: To compare the in vitro adherence of bovine lens epithelial cells (BLEC) onto the surface of intraocular lens (IOL) made of 4 different materials: polymethylmethacrylate (PMMA), silicone (SI), hydroxyethylmethacrylate (HEMA) and acrylic (Acrysof, ACRY). METHODS: BLEC were incubated with IOLs four 48 hours. Adherent cells were counted by flow cytometry technique. RESULTS: The amount of the adherent cells on IOLs of ACRY was the highest, that of HEMA the least, and there was no significant difference between the amount on PMMA and SI IOLs (P > 0.05). CONCLUSIONS: Adherence of lens epithelial cells to IOLs is influenced by the surface property of IOL materials, which may influence the occurrence of postoperative capsular opacification after IOL implantation. PMID- 11853627 TI - [Retinal detachment with anterior proliferative vitreoretinopathy and its surgical treatment]. AB - OBJECTIVE: To explore the clinical characteristics and surgical results of retinal detachment in eyes with anterior proliferative vitreoretinopathy (PVR). METHODS: 83 patients (83 eyes) of retinal detachment with grade C-D PVR were studied. Different surgical methods were used in treating anterior PVR and posterior PVR, respectively. RESULTS: Among the 83 patients, anterior PVR was in 34 eyes (41.0%) and posterior PVR 49 eyes (59.0%). The anterior PVR occurred in younger patients, mean age 28.5 years, more in male patients (85.3%) and more in grade D of PVR (55.9%); in the posterior PVR, the mean age was 38.9 years, 63.3% of patients were male and 24.5% of patients were in grade D of PVR. Eighty-three patients had a mean postoperative follow-up of 12.9 months. Complete retinal re attachment occurred in 24 eyes (70.6%) with anterior PVR and in 38 eyes with posterior PVR (77.6%; chi(2) = 0.51, P > 0.1). The final corrected visual acuity >or= 0.05 was in 11 eyes with anterior PVR (32.4%) and in 22 eyes with posterior PVR (44.9%; chi(2) = 1.32, P > 0.1). CONCLUSIONS: Eyes with anterior PVR have different clinical characteristics and worse surgical conditions when compared with eyes of posterior PVR. However, by the use of modern vitreoretinal surgery, the outcome in eyes with anterior PVR may become similar to that of posterior PVR. PMID- 11853628 TI - [An analysis of bacterial culture results in infectious endophthalmitis]. AB - OBJECTIVE: To study the changes of positive rate of bacterial culture of vitreous specimens from patients with infectious endophthalmitis during the past ten years and search for sensitive antibiotics for its treatment. METHODS: Vitreous specimens were taken from 304 eyes of 304 cases with clinically diagnosed infectious endophthalmitis for bacterial culture, and drug sensitivity test was carried out for the positive specimens. RESULTS: Bacterial positive rate was 53% with insignificant changes among the different years. Cefoperazone was the most sensitive antibiotic. CONCLUSION: Bacterial culture combined with drug sensitivity test of vitreous specimens is important in guiding the treatment of infectious endophthalmitis. PMID- 11853629 TI - [The clinical features and study on therapeutic effects of fungal keratitis]. AB - OBJECTIVE: To review the clinical features and therapeutic effects of fungal keratitis (FK). METHODS: 110 cases (110 eyes) with the disease were divided into two types: hyphomycetic keratitis in 94 eyes and candida keratitis in 16 eyes. The clinical features and therapeutic effects of the two types were observed and compared. RESULTS: The hyphomycetic keratitis generally had a plant traumatic history. The border of corneal infiltration was not clear with feather edges, and had a tendency to penetrate into the deep stroma of cornea. There were endothelial plaques, hypopyon and bad therapeutic effects. The pathogenicity of cardida keratitis (CK) was related to the local long-term use of glucocorticoid, the fungus seldom expanded into the deep corneal layers and infiltrated into the inner eye. Its therapeutic effects were better. The border of CK was characterized with limitations, and was clear. CONCLUSIONS: The clinical features of the two types are different. The clinical features caused by different or the same pathogenic fungi show great variations. Early diagnoses, correct use of medicine and operation are the key points to increase the curative rate. PMID- 11853630 TI - [Optical coherence tomography applied for measurement of nerve fiber layer thickness in normal eyes]. AB - OBJECTIVE: To evaluate the efficiency of optical coherence tomography (OCT) on quantifying retinal nerve fiber layer (RNFL) thickness and its reproducibility and investigate the difference of RNFL thickness between different groups. METHODS: According to their age, 152 eyes of 152 normal subjects were classified into six groups (such as 10 year, 20 year group, etc.), and investigated by OCT to measure the RNFL thickness. Each subject had circular scans around the optic nerve with a circle size of 3.46 mm. The RNFL thickness in different groups was analyzed. Intraclass correlation coefficient (ICC) in fifty eyes of 37 subjects was calculated. RESULTS: The mean RNFL thickness of 152 eyes was (90.09 +/- 10.81) microm, (140.27 +/- 10.60) microm, (85.03 +/- 14.01) microm, (140.27 +/- 9.70) microm and (114.11 +/- 6.08) microm (temporal, superior, nasal, inferior and the mean of the whole layer, respectively). Least significant difference (LSD)-t test was applied for multiple thickness comparisons of the mean and different quadrants among different groups. The results showed there were significant differences between the 60 year group and others except the 50 year group in temporal, superior, inferior quadrants and mean RNFL thickness (P = 0.000 - 0.019), and between each of 10 to 40 year groups and each of 50 to 60 year groups in the mean RFNL thickness (P = 0.000 - 0.026). ICCs of RNFL thickness measured by OCT were 0.78, 0.66, 0.64, 0.52 and 0.74 (temporal, superior, nasal, inferior and mean, respectively). CONCLUSIONS: Normal RNFL thickness can be measured accurately by OCT. There are some differences of RNFL thickness between different groups in normal persons. PMID- 11853631 TI - [Ultrastructure study on extraocular muscle of infiltrative exophthalmos]. AB - OBJECTIVE: To investigate histopathologic and ultrastructural changes of extraocular muscles (EOMs) in infiltrative exophthalmos in order to collect some evidences of pathogenesis. METHODS: Ten EOM samples in infiltrative exophthalmos were obtained after operation. Each of them was cut into two pieces. One of them was stained with HE and observed by light microscope. Another one was sectioned with ultrathin method and observed by transmission electron microscope. RESULTS: Under the light microscope, the extraocular muscular cells had no significant abnormality at the early stage, while at the middle and late stage, sarcoplasm coagulation, granular degeneration, vacuolization and necrosis were found in the cells. Under the electron microscope, there were disturbance and disappearance of the Z line in part of muscular fibers in early cases; at the middle and late stage, various degrees of sarcoplasmic reticulum dilatation, myofilament lysis and destruction with formation of vacuoles, secondary lysosome and remnant bodies were seen. In severe cases, the muscular cells could be completely destroyed and phagocytosed by macrophages, fibrosis occurred and myofibroblasts were found in some cases. CONCLUSIONS: The EOMs in infiltrative exophthalmos are destroyed in various degrees, and the muscular cells may be the target cells in Graves' ophthalmopathy. PMID- 11853632 TI - [Electron-immunostaining characteristics of platelet-derived growth factor, transforming growth factor beta1 and their receptors in epiretinal membranes]. AB - OBJECTIVE: To investigate the ultrastructural distribution of platelet-derived growth factor (PDGF), transforming growth factor beta(1) (TGF-beta(1)) and their receptors in epiretinal membranes (ERM) and discuss the clinical significance of the distribution. METHODS: 25 membrane specimens were surgically removed by vitrectomy from 9 patients with rhegmatogenous retinal detachment. Double staining techniques of immunoelectromicroscopy for eight antibodies (PDGF, TGF beta(1), PDGF receptor subunits alpha and beta, TGF-beta subunits I and II, type I and III collagen) were used in these specimens as previously designed. RESULTS: PDGF and TGF-beta(1) staining appeared to be stronger on early (< 2 months) and late (> 6 months) stage membranes than that of middle (2 - 6 months) stage of membranes, and the immunostaining intensity inverted with the degree of proliferative vitreoretinopathy (PVR). We found that the gold particles of PDGF and TGF-beta(1) tended to get together with that of type I and III collagen. In addition, the stainings of four growth factor receptors were frequently positive in a type of epithelial-like cells with rich cytoplasm components, and also the gold particles of PDGF and TGF-beta(1) were found around these cells. CONCLUSIONS: A concentration change of PDGF and TGF-beta(1) exists in development of ERM throughout. The epithelial-like cells are of a type of active cells, and autocrine and paracrine activity may be involved in ERM pathogenesis. PDGF and TGF-beta(1) influence the contractile activity of ERM. PMID- 11853633 TI - [The abnormal expression of epidermal growth factor receptor family in pterygium]. AB - OBJECTIVE: To evaluate the expression of type 1 growth factor receptor family [epidermal growth factor receptor (EGFR), ErbB2 and ErbB3] by the epithelium in pterygium so as to understand the pathogenesis of this disorder. METHODS: The immunofluorescent staining and Western blotting were performed in 15 patients with primary pterygium, compared to normal conjunctival epithelium, to determine the expression of EGFR, ErbB2 and ErbB3 proteins. RESULTS: The EGFR was present at the basal cells while the ErbB2 and ErbB3 were expressed by the superficial cells in normal conjunctival epithelium in immunofluorescent staining. Of 15 pterygia, 11 pterygia expressed the stains of EGFR, ErbB2 and ErbB3 in the full thickness of epithelium and with stronger staining compared to the control group, 4 of them showed the same staining pattern as that in the normal conjunctiva. Compared to normal conjunctiva, the stronger density of protein bands of EGFR, ErbB2 and ErbB3 was also demonstrated by western blotting in 11 pterygia with strong staining of these antibodies. CONCLUSIONS: The expression of EGFR, ErbB2 and ErbB3 is increased in pterygium, showing that the pterygium is a disorder with proliferation. The abnormal expression of EGFR, ErbB2 and ErbB3 by epithelium and their communicating with cytokines at stroma in pterygium may be one of the key pathogenetic factors in this disorder. PMID- 11853634 TI - [An ultrastructural observation on rat retina after photoreceptor cell implantation]. AB - OBJECTIVE: To further study the retinal neuronal signal. METHODS: The Wistar/RCS (RCS rats are rats with hereditary photoreceptor degeneration) rats were respectively as donors/acceptors, and the retinal pathway was reconstructed with the technique of pure photoreceptor transplantation. The photoreceptor layer of the retina was obtained with the technique of retinal whittle by manual method or excimer laser. The specimens were got separately at 2 weeks and 1 month after the transplantation and studied under the light and transmission electron microscopes. RESULTS: Most transplanted photoreceptors with physical poles were lined up regularly between the retinal pigment epithelium (RPE) and inner nuclear layer. It was shown that in the new outer plexiform layer the relatively integral synapse and its interconnection were seen. CONCLUSION: The retinal neuronal pathway can be reconstructed by retinal transplantation. PMID- 11853635 TI - [Glutamate in experimental acute elevated intraocular pressure models of rabbits]. AB - OBJECTIVE: To measure the concentration of glutamate (Glu) in the retina of eyes with experimentally elevated intraocular pressure (IOP). METHODS: High performance liquid chromatography (HPLC) was utilized to measure the Glu in the retina. 18 rabbits were divided into three groups, each group including 6 rabbits. In one eye of each rabbit in group 1, the IOP was elevated to 60 mm Hg (1 mm Hg = 0.133 kPa) by the anterior chamber perfusion with normal saline maintained for 4 hours, while the IOP of the other eye was maintained at 20 mm Hg by the same method. In group 2, the same experimental method was carried out, but the elevated IOP was maintained at 30 mm Hg. Group 3 was a normal control group without perfusion. RESULTS: High-IOP eyes (60 mm Hg and 30 mm Hg) had high glutamate concentration in the retinal tissue than that in low-IOP eyes (20 mm Hg) (P < 0.05). CONCLUSION: Elevated IOP induces higher content of glutamate in the retina. PMID- 11853637 TI - [Digital photography and blurred image processing of retinal nerve fiber layer in glaucoma]. AB - OBJECTIVE: Red-free photographs are effective in detecting retinal nerve fiber layer (RNFL). The reasons why it is not used by most clinics are difficulty in laborious developing and blurred image evaluating. To account for getting real time image and blurred image evaluating, the digital photography and computerized image processing are used. METHODS: RNFL photographs were taken by Nikon Fundus Camera connected with a high resolution digital camera (Kodak) and a green filter. As usual it is necessary to see the texture of RNFL in evaluating its image, but blurred images due to media opacities greatly lose their resolution. The way we used no focus on seeing the texture of RNFL, but on evaluating the distribution pattern of RNFL. RNFL images have more white reflex in temporal superior, temporal inferior, nasal superior and nasal inferior sections around optic disc according to the thickness of RNFL. Threshold segmentation was used, we shift a threshold bar to the position until the nasal or temporal section around optic disc is black with the software Photoshop. RESULTS: In the normal RNFL image, the white reflex was of butterfly pattern, i.e., the temporal superior and temporal inferior sections around optic disc were white. In cases with glaucoma, the characteristic of early damage of optic nerve was the loss of RNFL at the temporal superior and temporal inferior part of the retina, thus the normal butterfly pattern was changed, the temporal superior or/and temporal inferior sections were black. With this method it is more effective to evaluate RNFL. However, in cases with retinal pigment epithelium faint in color and in cases with choroidal defect, it is still difficult to evaluate the distribution of RNFL. CONCLUSIONS: It is an easy and effective way to detect glaucomatous damage with the digital photography and RNFL pattern evaluation. PMID- 11853636 TI - [Cultured bovine corneal epithelial cells express functional water channel]. AB - OBJECTIVE: To investigate the water permeability (P(f)) of the cultured bovine corneal epithelial cells (CBCEC) and whether aquaporins (AQPs) were present. METHODS: Laser light-scattering system was used to determine the volume change of CBCEC from isotonic status rapidly transferred to hypotonic one, and the P(f) value was calculated. Messenger RNA obtained from CBCEC was injected into Xenopus laevis oocytes. After four days, the P(f) was determined by monitoring volume changes of oocytes upon hypoosmotic challenge (180 mOsm/L changed to 15 mOsm/L). RESULTS: CBCEC challenged with 10% hypoosmotic solution at 37 degrees C swelled rapidly; the cellular P(f) was 72 microm/s. In the oocytes injected with mRNA from CBCEC, the P(f) increased to 76 microm/s. This increase was inhibited (by 89%) by 0.3 mmol/L HgCl(2). CONCLUSION: Our results suggest that CBCEC express a typical mercurial-sensitive functional water channel in good numbers, judging from the comparatively high P(f) we measured. These begin to reopen the question of the presence and extent of an epithelial role in control of corneal hydration. PMID- 11853638 TI - [Ultrasound biomicroscopic examination of secondary pupillary block glaucoma]. AB - OBJECTIVE: To study the imaging characteristics of ultrasound biomicroscopy of secondary pupillary block glaucoma. METHODS: 11 cases with pupillary block glaucoma diagnosed by ultrasound biomicroscopy were studied which had been suspected as malignant glaucoma at the beginning. RESULTS: There were no obviously typical characteristics of pupillary block glaucoma shown in the images of ultrasound biomicroscopy in these cases. However, the existence of posterior chamber in each case strongly supported the diagnosis of pupillary block glaucoma. The imaging characteristics of ultrasound biomicroscopy were different in terms of the different primary disorders which resulted in the pupillary block glaucoma including iridocyclitis, lens subluxation resulted from trauma and glaucoma filtration surgery. In the cases of lens subluxation, the main characteristic was the increase of the distance between the lens and the ciliary process at the site of subluxation and the lens forward movement. In the cases of iridocyclitis, anterior and posterior synechiae were the main characteristics. And in the cases of glaucoma filtration surgery there were two main characteristics. One was that the distance between the ciliary process and the equator of lens existed obviously, and the other was the occlusion of the inner ostium of glaucoma filtration surgery or peripheral iridectomy. CONCLUSION: Ultrasound biomicroscopy (UBM) has important value in the diagnosis of secondary pupillary block glaucoma and the differentiation between it and malignant glaucoma. Whether the posterior chamber exists or not is the differential point between the pupillary block glaucoma and the malignant glaucoma. PMID- 11853639 TI - [To screen, clone and sequence TIGR gene mutation in Chinese patients with primary open- angle glaucoma]. AB - OBJECTIVE: To study trabecular meshwork induced glucocorticoid response protein (TIGR) gene mutation in Chinese patients with primary open-angle glaucoma (POAG). METHODS: (1) From 70 patients with POAG and 20 normal controls, TIGR gene which consists of three exons (7 pairs of primer) was amplified by polymerase chain reaction (PCR). The mutation of PCR amplification products was evaluated by single-stranded conformation polymorphism (SSCP). (2) After the above PCR products were cloned into the PT-Adv vectors, the construction plasmids were evaluated by Ecor I endonuclease and direct sequence was performed. RESULTS: (1) With SSCP screening, the single strand band abnormality was found in the middle fragments of the third exon (the sixth pair of primer) of TIGR gene in two patients with POAG, but the control group appeared normal. (2) The two samples using clone and sequence showed that one had a 'GAT'- to- 'AAT' transition at amino acid Asp 338 Asn mutation; the base sequence of another one had no change. These results suggested that mutation rate of TIGR gene in Chinese patients with POAG be only 1.4% (1/70), being lower than that of foreigners. CONCLUSION: The pathogenesis of Chinese patients with POAG may be related with TIGR gene mutation, but the mutation rate is lower than that in foreigners, indicating that the mechanism of POAG in China and aboard is different and the pathogenesis of POAG has difference in regions and races. PMID- 11853640 TI - [Qualitative and quantitative measurement of retinal nerve fiber layer in primary open angle glaucoma by optical coherence tomography]. AB - OBJECTIVE: To study the characteristics of optical coherence tomography (OCT) of primary open angle glaucoma (POAG), the difference of retinal nerve fiber layer (RNFL) thickness between normal persons and patients with POAG and the correlation between RNLF and visual field index. METHODS: Eighty-three cases (149 eyes) with POAG and 83 normal persons (150 eyes) were tested by OCT with circular scans around the optic nerve head (diameter = 3.46 mm) to observe the features of OCT. Statistic analysis was taken to compare the difference of RNFL thickness in quadrants and means between normal and glaucomatous group, and the difference of the thickness among the stages in POAG. Linear correlation and regression analysis was used to show the correlation between RNFL thickness and visual field index of 115 eyes of glaucomatous patients. RESULTS: The RNFL thickness measured by OCT in normal subjects is the thickest in superior and inferior quadrants, less in thickness in temporal and the thinnest in nasal quadrant. The curve showed double peaks. The RNFL of glaucomatous patients showed local thinness or defect, diffuse thinness or combination of the above two types. The mean RNFL thickness of normal group was (90.1 +/- 10.8) microm, (140.4 +/- 10.5) microm, (85.2 +/- 14.0) microm, (140.4 +/- 9.7) microm and (114.2 +/- 6.0) microm, of glaucomatous group was (56.0 +/- 31.0) microm, (81.0 +/- 36.3) microm, (47.1 +/- 27.5) microm, (73.4 +/- 38.4) microm and (64.6 +/- 28.8) microm in temporal, superior, nasal, inferior quadrant and the whole area, respectively. There is significant difference of RNFL thickness between the normal and glaucomatous group (P < 0.000), and there are significant differences among the three stages (early, developing and late) of glaucomatous group (P < 0.000). There is a close negative relationship between RNFL thickness and visual field index (r = -0.796, P < 0.0001). The sensitivity and specificity of RNFL thickness measured by OCT were 93.3% and 92.0%, respectively. CONCLUSION: OCT can quantitatively measure the RNFL thickness and show the difference of RNFL between normal persons and glaucomatous patients. The RNFL thickness gradually decreases while visual field defect increases with the development of POAG. PMID- 11853641 TI - [The effect of three kinds of anti-glaucoma eyedrops on ocular blood flow]. AB - OBJECTIVE: Surveying the changes of ocular blood flow by continuous anti-glaucoma eyedrops for one week. METHODS: Thirty-two patients in glaucoma clinic were divided into three groups, one treated by 0.5% timolol, one by 0.5% levobunolol and the third by 0.1% dipivefrine, twice a day. Each patient got the retinal and choroidal angiography before and after the medication. We also made the intra- and inter-observer reproducibility test for verifying the credibility of the method used to determine the ocular blood flow. All the data were analyzed by Students't test. RESULTS: The reproducibility test revealed this method creditable. Timolol and levobunolol could decrease the choroidal blood flow significantly, there was a trend to decrease the optic nerve blood flow after continues application of timolol eyedrops. Dipivefrine did not induce any significant change on ocular blood flow except the mild artery constriction. CONCLUSION: The results suggest that it is necessary to be careful in choosing the anti-glaucoma eyedrops when ischemia plays an important role in glaucomatous damage. PMID- 11853642 TI - [Trabeculectomy combined with phacoemulsification for treatment of glaucoma complicated with cataract]. AB - OBJECTIVE: To evaluate the effectiveness of small-incision triple procedure, including phacoemulsification, posterior chamber intraocular lens implantation and trabeculectomy, in patients with coexisting glaucoma and cataract. METHODS: Twenty patients (26 eyes) with coexisting glaucoma and cataract underwent 3.5 mm incision triple procedure. The mean follow-up was 16.1 months (3 - 41 months). RESULTS: The mean preoperative intraocular pressure (IOP) was (23.01 +/- 2.63) mm Hg (1 mm Hg = 0.133 kPa) which decreased to a mean postoperative IOP of (13.93 +/ 1.85) mm Hg (P < 0. 001). Seventeen (65%) of 26 eyes had a best-corrected visual acuity of >or= 0.6 at the last follow-up (range, 0.05 - 1.0). The mean magnitude of astigmatism was 0.81 D (range, 0 - 3.00 D), 4 eyes had no astigmatism. Although 2 eyes used antiglaucoma medications shortly after the surgery, no eyes used any medications at the last follow-up. The early postoperative complications included corneal edema in 5 eyes (19%) and shallow anterior chamber in 3 eyes (12%). The late complication was mainly the after cataract in 6 eyes (23%). CONCLUSIONS: The small-incision triple procedure appears effective for treating selected patients with coexisting cataract and glaucoma, improving the valid visual acuity rapidly, reducing intraocular pressure with less medications and having less postoperative complications. PMID- 11853643 TI - [Phacoemulsification with silicone foldable intraocular lens implantation through a small incision in glaucomatous eyes after filtering surgery]. AB - OBJECTIVE: To investigate the method of cataract extraction in eyes that had undergone glaucoma filtering surgery and its effects on filtering bleb. METHODS: Phacoemulsification with posterior chamber intraocular lens implantation was performed on 262 eyes with a preexisting filtration bleb, in which at the temporal clear cornea a lateral or inferior lateral limbal incision was selected, the pupil sphincter was cut and the iris was sewed for the small and fixed pupil. RESULTS: The postoperative visual acuity of all patients improved in different degrees, with >or= 0.5 in 74.0% in the first month. The postoperative IOP increased by 3.08 mmHg. The functional filtering bleb was not apparently cicatrized. CONCLUSIONS: Glaucomatous eyes with cataract after filtering operation can rehabilitate their visual acuity and maintain the functional filtering bleb by phacoemulsification with foldable intraocular lens implantation through a temporal clear corneal incision or lateral scleral pocket incision. Theoretically, the effect of a transparent corneal incision on filtering bleb is less than that of a scleral pocket incision. PMID- 11853644 TI - [Corneal thickness and intraocular pressure in cases with ocular hypertension and glaucoma]. AB - OBJECTIVE: To investigate the relationship between central corneal thickness (CCT) and intraocular pressure (IOP) in patients with ocular hypertension (OH), normal tension glaucoma (NTG), primary open angle glaucoma (POAG) and normal subjects and the importance of CCT in the diagnosis of glaucoma. METHODS: The CCT was measured in 79 cases with NTG, 61 POAG, 73 OH and 50 normal subjects with an ultrasonic pachymeter. One eye in one subject randomly selected was used for inter-group comparison. The maximum IOP measured by Goldmann applanation tonometer without ocular hypotensive therapy during the follow-up was corrected with CCT by using Ehlers' method. RESULTS: The CCT of OH subjects (582 +/- 32) microm was significantly thicker than that of other groups (P < 0.001), while no differences were seen in CCT among normal, NTG and POAG groups, the thickness being (552 +/- 36) microm, (548 +/- 33) microm and (550 +/- 33) microm respectively. The corrected maximum pressure was significantly lower than that in the untreated ones in all the groups, and 0.05). The mean surgically-induced astigmatism at postoperative one day, one month and three months were (1.37 +/- 1.03), (1.24 +/- 0.93) and (1.04 +/- 0.75) D respectively. There was significant difference between the astigmatism at three months and at one day postoperatively (P < 0.05). CONCLUSIONS: The superior inversed frown shaped scleral small incision with no suture has little effect on corneal astigmatism. Vector analysis can systemically evaluate surgically induced astigmatism. The results of corneal topography are more reliable to reflect changes in corneal curvature than that of auto-ref-keratometer, and corneal topography has more important clinical value in evaluating surgically induced astigmatism. PMID- 11853650 TI - [The characteristics and curative effects of strabismus following retinal detachment surgery]. AB - OBJECTIVE: To describe the clinical characteristics and evaluate the surgical results in strabismus eyes which all had undergone retinal detachment surgery. METHODS: This study covered 24 strabismus cases (28 eyes) following retinal detachment operations. For these, examinations involving prism with alternative cover test or Krimsky test, eyeball movement, and diplopia analysis were performed. Strabotomy was carried out combined with or without scleral buckling removal or secondary intraocular lens implantation. The post-operative follow-up period ranged from 1 month to 24 months. RESULTS: (1) The main clinical feature in 16 strabismus cases (19 eyes) having managed by rhegmatogenous retinal detachment surgery was that the character of strabismus was related to the location of scleral explants. (2) Diplopia completely disappeared in 40.9% and partly disappeared in 54.5% of the cases. Strabismus was completely corrected in 63.6% of 27 eyes and partly corrected in 31.8% of them. CONCLUSIONS: (1) The strabismus following retinal detachment surgery is a restrictive strabismus, the clinical characteristics of which are different from that of palsy strabismus. (2) Strabotomy is effective in the treatment of restrictive strabismus. PMID- 11853649 TI - [Long-term observation of curative effects of posterior scleral reinforcement surgery in patients with juvenile progressive myopia]. AB - OBJECTIVE: To evaluate the long-term curative effects of the scleral reinforcement surgery in juvenile progressive myopia. METHOD: 25 patients (34 eyes) with the disease (average age, 17.12 +/- 7.47 years old) were observed for 3 to 5 years after posterior scleral reinforcement surgery. RESULTS: The naked visual acuities at postoperative one and three years were better than that before the operation (P < 0.01). The average axial elongation in the first, third and fifth year after the operation was 0.098 mm/year, 0.073 5 mm/year and 0.115 mm/year, respectively; the increment of refractive diopter was smaller than 0.50 D/year. CONCLUSION: The scleral reinforcement surgery is a safe and effective treatment for juvenile progressive myopia. PMID- 11853651 TI - [The measurement of normal values of exophthalmos, interpupillary distance and interorbital distance of children and adolescence in Xiamen and the rule of their development]. AB - OBJECTIVE: To investigate the developing rhythm of children and adolescent exophthalmos, interorbital distance (IOD) and interpupillary distance (IPD) and their mutual relationships. The study will provide biometry data for formulating juvenile criteria of correcting spectacles. METHODS: Random sampling was performed among children and adolescence aged 5 - 17 years in Xiamen. Adopting epidemic survey, we measured the biometry values of IOD, IPD and exophthalmos with a sliding gauge and exophthalmometer. RESULTS: The average value of exophthalmos was (14.48 +/- 1.71) mm. For the 5 - 8 years old children, the value of exophthalmos trended to increase with the increase of age, meanwhile the value of exophthalmos of adolescence aged 9 - 17 years old tended to be stabilized. The average value of IOD was (95.55 +/- 5.32) mm and the average value of IPD was (56.99 +/- 3.93) mm. Both the values of the IOD and IPD of the male were higher than that of the female. The results of the IOD and IDP distance increased with the increase of age and it trended to be periodic. There was positive correlation between the development of IOD and that of IPD, and it could be described by a linear equation. CONCLUSIONS: There is only one rush-increase stage of ocular development which is before the 8 years old in children. The rule of its development is in accordance with the growing and developing mode of neurological system. PMID- 11853652 TI - Treatment of Bacterial Meningitis. AB - The therapeutic approach to acute bacterial meningitis has changed in recent years as a result of changes in in vitro susceptibility of many meningeal pathogens to previously standard antimicrobial therapy. Given the emergence of strains of Streptococcus pneumoniae that are resistant to penicillin and the cephalosporins, the combination of vancomycin plus a third-generation cephalosporin is recommended as empiric therapy for suspected or proven pneumococcal meningitis, pending results of in vitro susceptibility testing. Strains of Neisseria meningitidis with reduced susceptibility to penicillin have also been described, although most patients with these resistant strains have recovered with standard penicillin therapy. Although the third-generation cephalosporins have greatly improved outcome in patients with meningitis caused by aerobic gram-negative bacilli, many organisms in this group are now resistant to these drugs; the carbapenems and fluoroquinolones may be effective alternative agents and have been successfully used in small case series. Further surveillance of the in vitro antimicrobial susceptibility patterns of meningeal pathogens is critical for future recommendations in the treatment of bacterial meningitis. PMID- 11853653 TI - The Oxazolidinones. AB - The need for effective antibiotics to manage the ever increasing frequency of antibiotic-resistant gram-positive infections in much of the developed world has led to the clinical development of the first oxazolidinone antibiotic, linezolid. Linezolid possesses bacteriostatic activity against both antibiotic-susceptible and resistant strains of staphylococci, enterococci, and streptococci of relevance to human infection. Clinical trials have confirmed its effectiveness in the treatment of serious infections of skin and soft tissue and the lower respiratory tract. Linezolid has also provided improved outcomes in the treatment of serious vancomycin-resistant enterococci and methicillin-resistant Staphylococcus aureus infections in a compassionate use program. Emergence of linezolid-resistant gram-positive cocci during clinical use has recently been described, suggesting that its present role in therapy should be reserved for the treatment of antibiotic-resistant gram-positive infections. PMID- 11853654 TI - Biological Warfare: Implications for Antimicrobial Use. AB - Biological warfare is intended to incapacitate a large number of individuals at a single exposure, creating epidemic-type disease, death, and social chaos. The organisms with potential for immediate use as bacteriologic weapons are Bacillus anthracis, Brucella melitensis, Yersinia pestis, and Vibrio cholera, all necessitating antibiotic therapy for a cure. It is reasonable, therefore, to assume that a biological attack, or even a hoax, would requiure thousands of individuals over a large area to begin antibiotic therapy. Issues such as antibiotic availability, logistical problems in antibiotic distribution, development of drug resistance, side effects influencing the individual, and adverse effects on the community due to the impact of mass therapy on the ecology, make biological warfare the most apocalyptic scenario for the creation of a "postantibiotic era." PMID- 11853656 TI - Clinical Trials Report. PMID- 11853655 TI - Chlamydia pneumoniae and Cardiovascular Disease. AB - Chlamydia pneumoniae, a respiratory pathogen, has been suggested as a risk factor for cardiovascular disease. Epidemiologic data are very controversial. Histopathologic and microbiologic studies have established an association between atherosclerosis and presence of C. pneumoniae, consistently finding C. pneumoniae DNA and antigens in atherosclerotic arteries. C. pneumoniae has been cultured from atherosclerotic arteries in several centers. An etiologic role for C. pneumoniae in initiation, acceleration of atherosclerosis, and/or acute ischemia remains debatable. In vitro studies have shown that C. pneumoniae can induce foam cell formation, low-density lipoprotein oxidation, and proinflammatory and procoagulant cytokine expression. Animal models of de novo initiation or enhancement of atherosclerosis have been developed. Preliminary trials of secondary prevention of coronary artery disease complications by antimicrobial agents show modest results. Better diagnostic tools, more diverse animal models, and clinical trials of primary prevention are needed. Meanwhile, results of ongoing large clinical trials on secondary prevention are eagerly awaited, but may not be definitive. PMID- 11853657 TI - Recent Advances and New Challenges in Travel Medicine. AB - Recent advances in travel medicine include the use of computer resources to obtain information on outbreaks and recommendations to travelers, the introduction of atovaquone/proguanil as chemoprophylaxis and treatment for malaria, the use of azithromycin as an alternative in the self-treatment of traveler's diarrhea, and the combination of hepatitis A and hepatitis B vaccines. At the same time, new challenges continue to appear. Shifts in the distribution of infections, such as West Nile virus and dengue fever, underscore the need for up-to-date information. Well-known infectious diseases, such as polio, meningococcal meningitis, and influenza are appearing in unexpected ways and settings. It is increasingly clear that travelers, while at risk for infections, also play a role in the global dispersal of pathogens, such as certain serogroups of Neisseria meningitidis and influenza. Increasing drug resistance affects the choice of drugs for treatment and chemoprophylaxis, and decisions about use of vaccines. Newly identified adverse events associated with yellow fever vaccine have prompted enhanced surveillance after vaccination and careful scrutiny of appropriate indications for the vaccine. PMID- 11853658 TI - An Update on the Geohelminths: Ascaris lumbricoides, Hookworms, Trichuris trichiura, and Strongyloides stercoralis. AB - Geohelminths remain prevalent throughout the developing world where levels of sanitation, personal hygiene, and maternal education are low. The five species of nematodes responsible for the bulk of disease are Ascaris lumbricoides, the hookworms Ancylostoma duodenale and Necator americanus, Trichuris trichiura, and Strongyloides stercoralis. Geohelminths are acquired through ingestion of fecally contaminated food or water or through contact with infected soil. In developing countries, infection with more than one nematode species and high worm burdens are common. The morbidity is substantial, particularly among children, and deaths occur. Geohelminthic infections are encountered in industrialized countries among immigrants and long-term travelers who have lived in endemic regions where sanitation is poor, and occasionally following autochthonous transmission. PMID- 11853659 TI - Update on Lymphatic Filarial Infections. AB - Filarial infections remain significant causes of disability in tropical areas worldwide. However, insights into the developmental and molecular biology of the parasite and the immunobiology of the host response to infection have advanced our understanding, even as progress is being made towards implementing eradication programs. This article summarizes some of the recent advances in the understanding of filarial biology and parasite immune evasion mechanisms, and reviews those newer aspects of diagnosis and treatment most relevant to clinicians. PMID- 11853661 TI - Treatment of Primary HIV. AB - Primary HIV-1 infection refers to the events surrounding the acquisition of HIV-1 infection. It is commonly associated with a nonspecific clinical syndrome that occurs within 2 to 4 weeks after exposure in 40% to 90% of persons acquiring HIV 1. Patients identified prior to seroconversion often have plasma titers in excess of 500,000 copies/mL. Over time, plasma HIV-1 RNA titers decrease and eventually reach a "set point." Treatment of primary HIV-1 infection with highly active antiretroviral therapy does not prevent establishment of chronic infection. However, it potentially may decrease the viral set point, prevent evolution of resistant mutants, preserve immune function, improve clinical outcome, and possibly allow for viral control after withdrawal of antiretroviral therapy. Transmission of viral strains with decreased susceptibility to antiviral drugs increases the difficulty of choosing an antiretroviral regimen. Other medications, including immunomodulators, are under study as adjuvant therapy for treatment of primary HIV-1 infection. PMID- 11853660 TI - Update on Traveler's Diarrhea. AB - Diarrhea is one of the most common health problems among travelers. Although enterotoxigenic Escherichia coli is implicated most commonly, enteroaggregative E. coli has recently been described as a major pathogen. Shigella, Campylobacter, and Salmonella organisms are less common causes of acute diarrhea, and intestinal protozoa are typical causes of protracted diarrhea. Although education is the mainstay of prevention measures, behavior modification has been shown to be difficult. Chemoprevention is rarely required with the availability of effective treatment, but there has been some interest in the use of vaccines. Maintenance of hydration is most important in children. In addition to bismuth preparations and loperamide, newer agents being developed for symptomatic relief include zaldaride maleate and racecadotril. Fluoroquinolones effectively treat severe traveler's diarrhea, and even a single dose may be sufficient. However, with the emergence of resistance, particularly in Campylobacter infection, other agents are required; interest has focused on azithromycin and rifaximin. PMID- 11853662 TI - Virologic and Immunologic Response to Highly Active Antiretroviral Therapy. AB - Highly active antiretroviral therapy (HAART) delays clinical progression to AIDS by suppressing viral replication, allowing the immune system to reconstitute. These virologic and immunologic consequences do not occur uniformly among HAART users; markers of HIV disease stage at the time of HAART initiation are critical determinants of the progression while under HAART. In this paper, we review studies describing the heterogeneous virologic and immunologic progression following the initiation of HAART, and update findings obtained in the Multicenter AIDS Cohort Study that show that CD4 cell count and history of antiretroviral therapy at the time of initiation are independent determinants of response. PMID- 11853663 TI - No DREAM, No pain. Closing the spinal gate. AB - Pain transmission in the spinal cord is regulated by a balance of facilitatory and inhibitory influences operating on the neural circuits of the somatosensory system. The transcriptional repressor DREAM acts constitutively to suppress prodynorphin expression in spinal cord neurons. Knocking out DREAM results in sufficient dynorphin expression to produce a strong reduction in generalized pain behavior, highlighting the role that intracellular molecules play in modulating pain gating in the spinal cord. PMID- 11853664 TI - A tense time for the nuclear envelope. AB - When many cells divide, the nuclear envelope poses a problem: the spindle microtubules can't access the chromosomes. Two recent papers in Cell describe how the spindle solves this problem by literally pulling open the nucleus at the beginning of mitosis. PMID- 11853665 TI - Origins of human virus diversity. AB - The diverse consequences of different viral infections on the human population reflect both the history of their origins and the nature of their ongoing evolution. Advances in the ease of finding and characterizing viral genome sequences are having a major impact on our understanding of the diversity of human viruses and of their relatives infecting other species. PMID- 11853666 TI - Don C. Wiley (1944-2001). PMID- 11853667 TI - Evidence that the Ipl1-Sli15 (Aurora kinase-INCENP) complex promotes chromosome bi-orientation by altering kinetochore-spindle pole connections. AB - How sister kinetochores attach to microtubules from opposite spindle poles during mitosis (bi-orientation) remains poorly understood. In yeast, the ortholog of the Aurora B-INCENP protein kinase complex (Ipl1-Sli15) may have a role in this crucial process, because it is necessary to prevent attachment of sister kinetochores to microtubules from the same spindle pole. We investigated IPL1 function in cells that cannot replicate their chromosomes but nevertheless duplicate their spindle pole bodies (SPBs). Kinetochores detach from old SPBs and reattach to old and new SPBs with equal frequency in IPL1+ cells, but remain attached to old SPBs in ipl1 mutants. This raises the possibility that Ipl1-Sli15 facilitates bi-orientation by promoting turnover of kinetochore-SPB connections until traction of sister kinetochores toward opposite spindle poles creates tension in the surrounding chromatin. PMID- 11853668 TI - Specific ablation of Stat3beta distorts the pattern of Stat3-responsive gene expression and impairs recovery from endotoxic shock. AB - Alternative splicing of the gene for Stat3, a transcription factor activated by the IL-6 family of cytokines, produces two isoforms: Stat3alpha and a dominant negative variant, Stat3beta. Stat3beta-deficient mice were generated by gene targeting. Despite intact expression and phosphorylation of Stat3alpha, overall Stat3 activity was impaired in Stat3beta(-/-) cells. Global comparison of transcription in Stat3beta(+/+) and Stat3beta(-/-) cells revealed stable differences. Stat3beta-deficient mice exhibit diminished recovery from endotoxic shock and hyperresponsiveness of a subset of endotoxin-inducible genes in liver. The hepatic response to endotoxin in wild-type mice is accompanied by a transient increase in the ratio of Stat3beta to Stat3alpha. These findings indicate a critical role for Stat3beta in the control of systemic inflammation. PMID- 11853669 TI - Structural basis for E2-mediated SUMO conjugation revealed by a complex between ubiquitin-conjugating enzyme Ubc9 and RanGAP1. AB - E2 enzymes catalyze attachment of ubiquitin and ubiquitin-like proteins to lysine residues directly or through E3-mediated reactions. The small ubiquitin-like modifier SUMO regulates nuclear transport, stress response, and signal transduction in eukaryotes and is essential for cell-cycle progression in yeast. In contrast to most ubiquitin conjugation, the SUMO E2 enzyme Ubc9 is sufficient for substrate recognition and lysine modification of known SUMO targets. Crystallographic analysis of a complex between mammalian Ubc9 and a C-terminal domain of RanGAP1 at 2.5 A reveals structural determinants for recognition of consensus SUMO modification sequences found within SUMO-conjugated proteins. Structure-based mutagenesis and biochemical analysis of Ubc9 and RanGAP1 reveal distinct motifs required for substrate binding and SUMO modification of p53, IkappaBalpha, and RanGAP1. PMID- 11853670 TI - Vacuole fusion at a ring of vertex docking sites leaves membrane fragments within the organelle. AB - Three membrane microdomains can be identified on docked vacuoles: "outside" membrane, not in contact with other vacuoles, "boundary" membrane that contacts adjacent vacuoles, and "vertices," where boundary and outside membrane meet. In living cells and in vitro, vacuole fusion occurs at vertices rather than from a central pore expanding radially. Vertex fusion leaves boundary membrane within the fused organelle and is an unexpected pathway for the formation of intralumenal membranes. Proteins that regulate docking and fusion (Vac8p, the GTPase Ypt7p, its HOPS/Vps-C effector complex, the t-SNARE Vam3p, and protein phosphatase 1) accumulate at these vertices during docking. Their vertex enrichment requires cis-SNARE complex disassembly and is thus part of the normal fusion pathway. PMID- 11853671 TI - A chemical-genetic strategy implicates myosin-1c in adaptation by hair cells. AB - Myosin-1c (also known as myosin-Ibeta) has been proposed to mediate the slow component of adaptation by hair cells, the sensory cells of the inner ear. To test this hypothesis, we mutated tyrosine-61 of myosin-1c to glycine, conferring susceptibility to inhibition by N(6)-modified ADP analogs. We expressed the mutant myosin-1c in utricular hair cells of transgenic mice, delivered an ADP analog through a whole-cell recording pipette, and found that the analog rapidly blocked adaptation to positive and negative deflections in transgenic cells but not in wild-type cells. The speed and specificity of inhibition suggests that myosin-1c participates in adaptation in hair cells. PMID- 11853672 TI - Crystal structure of sTALL-1 reveals a virus-like assembly of TNF family ligands. AB - TALL-1/BAFF/BLyS was recently identified as a member of the tumor necrosis factor (TNF) ligand family. The crystal structure of the functional soluble TALL-1 (sTALL-1) has been determined at 3.0 A. sTALL-1 forms a virus-like assembly with 200 A diameter in the crystals, containing 60 sTALL-1 monomers. The cluster formation is mediated by a "flap" region of the sTALL-1 monomer. The virus-like assembly was also detected in solution using gel filtration and electron microscopy. Deletion of the flap region disrupted the formation of the virus-like assembly. The mutant sTALL-1 still bound its receptor but could not activate NF kappaB and did not stimulate B lymphocyte proliferation. Finally, we found the virus-like cluster of sTALL-1 exists in physiological condition. We propose that this virus-like assembly of sTALL-1 is the functional unit for TALL-1 in vivo. PMID- 11853673 TI - Kre1p, the plasma membrane receptor for the yeast K1 viral toxin. AB - Saccharomyces cerevisiae K1 killer strains are infected by the M1 double-stranded RNA virus encoding a secreted protein toxin that kills sensitive cells by disrupting cytoplasmic membrane function. Toxin binding to spheroplasts is mediated by Kre1p, a cell wall protein initially attached to the plasma membrane by its C-terminal GPI anchor. Kre1p binds toxin directly. Both cells and spheroplasts of Deltakre1 mutants are completely toxin resistant; binding to cell walls and spheroplasts is reduced to 10% and < 0.5%, respectively. Expression of K28-Kre1p, an inactive C-terminal fragment of Kre1p retaining its toxin affinity and membrane anchor, fully restored toxin binding and sensitivity to spheroplasts, while intact cells remained resistant. Kre1p is apparently the toxin membrane receptor required for subsequent lethal ion channel formation. PMID- 11853674 TI - Integrin alpha3beta1 (CD 49c/29) is a cellular receptor for Kaposi's sarcoma associated herpesvirus (KSHV/HHV-8) entry into the target cells. AB - Human herpesvirus-8 (HHV-8) is implicated in the pathogenesis of Kaposi's sarcoma. HHV-8 envelope glycoprotein B possesses the RGD motif known to interact with integrin molecules, and HHV-8 infectivity was inhibited by RGD peptides, antibodies against RGD-dependent alpha3 and beta1 integrins, and by soluble alpha3beta1 integrin. Expression of human alpha3 integrin increased the infectivity of virus for Chinese hamster ovary cells. Anti-gB antibodies immunoprecipitated the virus-alpha3 and -beta1 complexes, and virus binding studies suggest a role for alpha3beta1 in HHV-8 entry. Further, HHV-8 infection induced the integrin-mediated activation of focal adhesion kinase (FAK). These findings implicate a role for alpha3beta1 integrin and the associated signaling pathways in HHV-8 entry into the target cells. PMID- 11853675 TI - Molecular basis for species-specific sensitivity to "hot" chili peppers. AB - Chili peppers produce the pungent vanilloid compound capsaicin, which offers protection from predatory mammals. Birds are indifferent to the pain-producing effects of capsaicin and therefore serve as vectors for seed dispersal. Here, we determine the molecular basis for this species-specific behavioral response by identifying a domain of the rat vanilloid receptor that confers sensitivity to capsaicin to the normally insensitive chicken ortholog. Like its mammalian counterpart, the chicken receptor is activated by heat or protons, consistent with the fact that both mammals and birds detect noxious heat and experience thermal hypersensitivity. Our findings provide a molecular basis for the ecological phenomenon of directed deterence and suggest that the capacity to detect capsaicin-like inflammatory substances is a recent acquisition of mammalian vanilloid receptors. PMID- 11853677 TI - Anchor-dependent lipofection with non-glycerol based cytofectins containing single 2-hydroxyethyl head groups. AB - Detailed structure-activity investigations aimed at probing the anchor chain length dependency for glycerol-based lipofectins have been reported previously. Herein, we report on the first detailed investigation on the anchor-dependent transfection biology of non-glycerol based simple monocationic cytofectins containing single 2-hydroxyethyl head group functionality using 11 new structural analogs of our previously published first generation of non-glycerol based transfection lipids (lipids 1-11). The C-14 and C-16 analogs of DOMHAC (lipids 4 and 5, respectively) were found to be remarkably efficient in transfecting COS-1 cells. In addition, the present anchor-dependency investigation also revealed that the C-14 analog of DOHEMAB (lipid 10) is significantly efficient in transfecting both COS-1 and NIH3T3 cells. Our results also indicate that too strong lipid-DNA interactions might result in weaker transfection for non glycerol based cationic lipids. In summary, the anchor-dependence investigations presented here convincingly demonstrate that non-glycerol based cationic lipids containing a single hydroxyethyl head group and hydrophobic C-14 or C-16 anchors are promising non-toxic cationic transfection lipids for future use in liposomal gene delivery. PMID- 11853678 TI - Conformational mapping of the N-terminal peptide of HIV-1 gp41 in membrane environments using (13)C-enhanced Fourier transform infrared spectroscopy. AB - The N-terminal domain of HIV-1 glycoprotein 41000 (FP; residues 1--23; AVGIGALFLGFLGAAGSTMGARSCONH(2)) participates in fusion processes underlying virus -cell infection. Here, we use physical techniques to study the secondary conformation of synthetic FP in aqueous, structure-promoting, lipid and biomembrane environments. Circular dichroism and conventional, (12)C-Fourier transform infrared (FTIR) spectroscopy indicated the following alpha-helical levels for FP in 1-palmitoyl-2-oleoylphosphatidylglycerol (POPG) liposomes hexafluoroisopropanol (HFIP)>trifluoroethanol (TFE)>phosphate-buffered saline (PBS). (12)C-FTIR spectra also showed disordered FP structures in these environments, along with substantial beta-structures for FP in TFE or PBS. In further experiments designed to map secondary conformations to specific residues, isotope-enhanced FTIR spectroscopy was performed using a suite of FP peptides labeled with (13)C-carbonyl at multiple sites. Combining these (13)C-enhanced FTIR results with molecular simulations indicated the following model for FP in HFIP: alpha-helix (residues 3-16) and random and beta-structures (residues 1-2 and residues 17-23). Additional (13)C-FTIR analysis indicated a similar conformation for FP in POPG at low peptide loading, except that the alpha-helix extends over residues 1-16. At low peptide loading in either human erythrocyte ghosts or lipid extracts from ghosts, (13)C-FTIR spectroscopy showed alpha helical conformations for the central core of FP (residues 5-15); on the other hand, at high peptide loading in ghosts or lipid extracts, the central core of FP assumed an antiparallel beta-structure. FP at low loading in ghosts probably inserts deeply as an alpha-helix into the hydrophobic membrane bilayer, while at higher loading FP primarily associates with ghosts as an aqueous-accessible, beta sheet. In future studies, (13)C-FTIR spectroscopy may yield residue-specific conformations for other membrane-bound proteins or peptides, which have been difficult to analyze with more standard methodologies. PMID- 11853679 TI - Mutual inhibition of the dimerized Na/Ca-K exchanger in rod photoreceptors. AB - In the dark, rod photoreceptors sustain a continuous influx of Na and Ca ions through the cGMP-gated channels of the rod outer segments (ROS). Whereas Na ions are extruded in the inner segment by the Na-pump, Ca ions are extruded already in the ROS by Na/Ca-K exchange. Our previous findings indicate that in the ROS plasma membrane, exchanger and channel form a complex of two exchangers associated per channel. Here, we report evidence of a novel regulatory mechanism of the dimerized exchanger, based on the following findings: (1), thiol-specific cross-linking with dimaleimides resulted in an increase of the Na/Ca-K exchange activity which correlated with the size of the cross-linking reagent, i.e., with increasing separation of the monomers in a dimerized exchanger; (2), partial proteolysis of the exchanger also increased the exchange rate by about a factor of two; (3), disintegration of the channel-exchanger complex by solubilization of the ROS membranes and preparation of proteoliposomes resulted in a twofold enhancement of the exchange rate; however (4), partial proteolysis of proteoliposomes, in which the exchanger molecules exist as monomers, did not result in any enhancement of the exchange rate. These findings suggest an inhibitory protein domain at the contact site of the dimerized exchanger. The physiological implication of this inference will be discussed in terms of a potential allosteric regulation of the exchanger in the channel-exchanger complex. PMID- 11853680 TI - Role of the choline exchanger in Na(+)-independent Mg(2+) efflux from rat erythrocytes. AB - Two types of Na(+)-independent Mg(2+) efflux exist in erythrocytes: (1) Mg(2+) efflux in sucrose medium and (2) Mg(2+) efflux in high Cl(-) media such as KCl-, LiCl- or choline Cl-medium. The mechanism of Na(+)-independent Mg(2+) efflux in choline Cl medium was investigated in this study. Non-selective transport by the following transport mechanisms has been excluded: K(+),Cl(-)- and Na(+),K(+),Cl( )-symport, Na(+)/H(+)-, Na(+)/Mg(2+)-, Na(+)/Ca(2+)- and K(+)(Na(+))/H(+) antiport, Ca(2+)-activated K(+) channel and Mg(2+) leak flux. We suggest that, in choline Cl medium, Na(+)-independent Mg(2+) efflux can be performed by non selective transport via the choline exchanger. This was supported through inhibition of Mg(2+) efflux by hemicholinum-3 (HC-3), dodecyltrimethylammonium bromide (DoTMA) and cinchona alkaloids, which are inhibitors of the choline exchanger. Increasing concentrations of HC-3 inhibited the efflux of choline and efflux of Mg(2+) to the same degree. The K(d) value for inhibition of [(14)C]choline efflux and for inhibition of Mg(2+) efflux by HC-3 were the same within the experimental error. Inhibition of choline efflux and of Mg(2+) efflux in choline medium occurred as follows: quinine>cinchonine>HC-3>DoTMA. Mg(2+) efflux was reduced to the same degree by these inhibitors as was the [(14)C]choline efflux. PMID- 11853681 TI - Modes of membrane interaction of a natural cysteine-rich peptide: viscotoxin A3. AB - Among the very homologous family of alpha- and beta-thionins, known for their antimicrobial activity, the viscotoxin subfamily differs from other members because it is cytotoxic against tumoral cells but weakly hemolytic. We studied the interactions between the most active of these toxins, viscotoxin A3 (VA3), and model membranes made of phosphatidylcholine and phosphatidylserine (PS), the major zwitterionic and acidic phospholipids found in eukaryotic cells. Monolayer studies showed that electrostatic forces are essential for the interaction and are mainly involved in modulating the embedding of the toxin in the PS head group region. This in turn induces membrane stiffening, as shown by fluorescence polarization assays with 1,6-diphenyl-1,3,5-hexatriene and its derivatives. Moreover, vesicle permeabilization analyses showed that there are two modes of interaction, which are directly related to the stiffening effect and depend on the amount of VA3 bound to the surface of the vesicles. We propose an interaction model in which the embedding of VA3 in the membrane induces membrane defects leading to the gradual release of encapsulated dye. When the surfaces of the vesicles are saturated with the viscotoxin, complete vesicle destabilization is induced which leads to bilayer disruption, all-or-none encapsulated dye release and rearrangement of the vesicles. PMID- 11853682 TI - Translocation of protegrin I through phospholipid membranes: role of peptide folding. AB - The protegrin PG-1, belonging to the family of beta-stranded antimicrobial peptides, exerts its activity by forming pores in the target biological membranes. Linear analogues derived from PG-1 do not form pores in the phospholipid membranes and have been used successfully to deliver therapeutic compounds into eucaryotic cells. In this paper, the translocation of PG-1 and of a linear analogue through artificial phospholipid membranes was investigated, leading to a possible mechanism for the activity of these peptidic vectors. We report here that [12W]PG-1, a fluorescent analogue of PG-1, is able to translocate through lipid bilayers and we demonstrate that this property depends on its secondary structure. Our results agree with the recent mechanism proposed for the translocation and permeabilisation activities of several helical and beta stranded peptides. In addition, our data corroborate recent work suggesting that certain protegrin-derived vectors enter into endothelial cells by adsorptive mediated endocytosis. PMID- 11853684 TI - Regulation of jejunal glucose transporter expression by forskolin. AB - We have investigated the effects of forskolin on enterocyte membrane expression of the glucose transporters, SGLT1 and GLUT2, which are thought to be the main entry and efflux pathways for glucose, respectively. Forskolin treatment increased SGLT1 but decreased GLUT2 expression in mid and lower villus enterocytes. No change in transporter expression was noted in upper villus cells. Likewise, cyclic AMP levels were raised in mid and lower but not upper villus cells. The implications of these data for glucose transport are discussed. PMID- 11853683 TI - Inhibition of Mrp2- and Ycf1p-mediated transport by reducing agents: evidence for GSH transport on rat Mrp2. AB - Mammalian Mrp2 and its yeast orthologue, Ycf1p, mediate the ATP-dependent cellular export of a variety of organic anions. Ycf1p also appears to transport the endogenous tripeptide glutathione (GSH), whereas no ATP-dependent GSH transport has been detected in Mrp2-containing mammalian plasma membrane vesicles. Because GSH uptake measurements in isolated membrane vesicles are normally carried out in the presence of 5-10 mM dithiothreitol (DTT) to maintain the tripeptide in the reduced form, the present study examined the effects of DTT and other sulfhydryl-reducing agents on Ycf1p- and Mrp2-mediated transport activity. Uptake of S-dinitrophenyl glutathione (DNP-SG), a prototypic substrate of both proteins, was measured in Ycf1p-containing Saccharomyces cerevisiae vacuolar membrane vesicles and in Mrp2-containing rat liver canalicular plasma membrane vesicles. Uptake was inhibited in both vesicle systems in a concentration-dependent manner by DTT, dithioerythritol, and beta mercaptoethanol, with concentrations of 10 mM inhibiting by approximately 40%. DTT's inhibition of DNP-SG transport was noncompetitive. In contrast, ATP dependent transport of [(3)H]taurocholate, a substrate for yeast Bat1p and mammalian Bsep bile acid transporters, was not significantly affected by DTT. DTT also inhibited the ATP-dependent uptake of GSH by Ycf1p. As the DTT concentration in incubation solutions containing rat liver canalicular plasma membrane vesicles was gradually decreased, ATP-dependent GSH transport was now detected. These results demonstrate that Ycf1p and Mrp2 are inhibited by concentrations of reducing agents that are normally employed in studies of GSH transport. When this inhibition was partially relieved, ATP-dependent GSH transport was detected in rat liver canalicular plasma membranes, indicating that both Mrp2 and Ycf1p are able to transport GSH by an ATP-dependent mechanism. PMID- 11853685 TI - Is inhibition of cyclooxygenase required for the anti-tumorigenic effects of nonsteroidal, anti-inflammatory drugs (NSAIDs)? In vitro versus in vivo results and the relevance for the prevention and treatment of cancer. AB - Active research is being conducted to unravel the cellular mechanisms mediating the anti-tumorigenic effects of nonsteroidal anti-inflammatory drugs (NSAIDs) and their association with cyclooxygenase (COX) inhibition. The majority of NSAIDs inhibit either COX-1, COX-2, or both and exert their anti-COX, anti-inflammatory, and anti-tumorigenic effects in vivo in a parallel dose-dependent manner. The effects are seen at NSAID blood plasma concentrations of 0.1-5 microM. Significantly, the same compounds tested at the same concentrations in incubations with cultured tumor cells in vitro similarly inhibit COX activities but are devoid of anti-proliferative activity. Yet, at much higher concentrations (100-20,000 microM), these same NSAIDs do exert anti-proliferative effects in vitro due to apparent non-specific toxic effects, as evidenced by disruption of ion transport and mitochondrial oxidation in some cells. A small group of NSAIDs (e.g. sulindac) do not inhibit COX enzymes significantly but can reduce the synthesis of prostanoids by alternate mechanisms. One such mechanism is inhibition of agonist-stimulated phospholipase-mediated release of arachidonic acid from phospholipids leading to depressed synthesis of prostanoids, especially prostaglandin E(2) (PGE(2)). Another group of non-COX inhibitors are the R isomers of NSAIDs, based on the structure of 2-arylpropionic acid. These compounds exert anti-proliferative effects in vivo, acting by an as yet undetermined mechanism. A possible caveat in these data is an R to S chiral transformation in vivo that would render the R-isomer effect as being due to the S-isomer generated in vivo from it. Demonstration of minimal or no R to S inversion under the experimental in vivo conditions employed is, therefore, a necessary control in these studies. The overall body of data supports the conclusion that, for COX-inhibiting NSAIDs, their anti-tumorigenic effect in vivo is due to, and depends upon, inhibition of tumor COX enzymes, primarily COX-2. The cellular effects seen when adding high concentrations of NSAIDs to tumor cells cultured in vitro and the mechanisms proposed to mediate these effects may not have substantial relevance to the mechanisms that mediate the effects of NSAIDs in vivo. PMID- 11853686 TI - Intracellular mechanisms mediating the inhibition of apoB-containing lipoprotein synthesis and secretion in HepG2 cells by avasimibe (CI-1011), a novel acyl coenzyme A: cholesterol acyltransferase (ACAT) inhibitor. AB - We have studied the cellular and molecular mechanisms involved in the suppression of apoB secretion from HepG2 cells following incubation with avasimibe (CI-1011), a novel inhibitor of acyl-coenzyme A: cholesterol acyltransferase (ACAT). Cellular lipid analysis revealed that avasimibe significantly decreased the synthesis of cholesterol and cholesteryl ester, and, at higher doses, of triglyceride. Time-course trypsin protection assays revealed that avasimibe induced the accumulation of translocationally arrested apoB intracellularly. Pulse-chase studies showed that the treatment with avasimibe induced a >75% decrease in apoB secretion relative to control, but initially enhanced the protein stability and cellular accumulation of apoB. Subcellular fractionation of microsomes further confirmed the accumulation of secretion-incompetent apoB lipoproteins in the endoplasmic reticulum (ER) and Golgi compartments of avasimibe-treated HepG2 cells. Although incubation of drug-treated cells with carbobenzoxyl-leucinyl-leucinyl-leucinal (MG132), a potent proteasome inhibitor, increased cellular apoB (70%), it failed to increase apoB secretion. Drug treatment induced an accumulation of secretion-incompetent apoB-containing lipoprotein particles, the majority of which demonstrated a density in a range similar to that of high-density lipoprotein. However, studies in permeabilized cells demonstrated that, at longer chase times, intracellularly accumulated apoB was eventually degraded, indicating that the inhibition of degradation may be transient. Oleate treatment of avasimibe-treated cells partially restored apoB secretion but not to the levels seen in control cells. In summary, we hypothesize that avasimibe acutely blocks the secretion of apoB and its associated lipoproteins from HepG2 cells, transiently enhancing its membrane association and cellular accumulation with eventual intracellular degradation of accumulated apoB. PMID- 11853687 TI - Identification of alpha-dicarbonyl scavengers for cellular protection against carbonyl stress. AB - Tissue deterioration and aging have long been associated with the accumulation of chemically induced protein and DNA damage. Reactive oxygen species (ROS) and reactive carbonyl species (RCS), especially alpha-dicarbonyl compounds, are key mediators of damage caused by oxidative stress, glycation, and UV-irradiation. The toxic effects of ROS are counteracted in vivo by antioxidants and antioxidant enzymes, and the deleterious effects of one RCS, methylglyoxal, are counteracted by a ubiquitous glyoxalase system. Carbonyl stress as a result of toxic effects of various mono-dicarbonyls (e.g. 4-hydroxynonenal) and alpha-dicarbonyls (e.g. glyoxal and deoxyosones) cannot be directly antagonized by antioxidants, and only a small number of biological carbonyl scavengers like glutathione (GSH) have been identified to date. We have developed a new screening method for the identification of carbonyl scavengers using a rapid glycation system that proceeds independent of oxygen and therefore, excludes identification of inhibitory compounds acting as antioxidants. Using this screening assay adapted to 96-well microtiter plates, we have identified the cysteine derivative 3,3 dimethyl-D-cysteine as a potent inhibitor of non-oxidative advanced glycation. Comparative kinetic analyses demonstrated the superior alpha-oxoaldehyde scavenging activity of D-penicillamine over that of aminoguanidine. D Penicillamine traps alpha-oxoaldehydes by forming a 2-acylthiazolidine derivative as shown by structure elucidation of reaction products between D-penicillamine and methylglyoxal or phenylglyoxal. We demonstrated that upon co-incubation, D penicillamine protects human skin keratinocytes and fibroblasts (CF3 cells) against glyoxal- and methylglyoxal-induced carbonyl toxicity. Our research qualifies alpha-amino-beta-mercapto-beta,beta-dimethyl-ethane as a promising pharmacophore for the development of related alpha-dicarbonyl scavengers as therapeutic agents to protect cells against carbonyl stress. PMID- 11853688 TI - Vanadyl as a catalyst of human lipoprotein oxidation. AB - Lipoprotein oxidation, which is relevant to atherogenesis, can be induced by redox-active transition metals, such as copper. Vanadium is a metal usually used as vanadyl to improve metabolic control in diabetic patients; given its redox active properties, we have investigated possible oxidative effects of the metal on lipoproteins from healthy and diabetic subjects. Beginning from 10 microM, vanadyl, but not vanadate, induced oxidation of the non-HDL fraction, which was inhibited by EDTA, butylated hydroxytoluene and Vitamins E and C, but not by mannitol, SOD and catalase. Differently from copper, vanadyl could oxidize directly lipoprotein lipids, although it showed a lower oxidant activity against critical tryptophan residues of the lipoprotein protein moiety. Moreover, the non HDL fraction of diabetic patients was more susceptible to vanadyl-dependent oxidation than that of controls. Thus, vanadium, in its reduced form which may be used in humans, can oxidize the non-HDL fraction through oxidative effects exerted especially on lipoprotein lipids; the specific pro-oxidant activity of vanadyl is more evident with lipoproteins of diabetic patients. Given also the tissue accumulating capacity of vanadium conceivably in a reduced form, its prolonged administration to humans, especially to diabetic patients without adequate antioxidant supplementation, needs caution. PMID- 11853689 TI - Methionine synthase activity and sulphur amino acid levels in the rat liver tumour cells HTC and Phi-1. AB - Methionine dependence has been reported in tumour cells and suggested as a possible target for chemotherapeutic drugs. The underlying defect has not been extensively researched, nor have levels of sulphur amino acids been examined in these cells. This study compared two rat liver tumour cell lines. One was found to be methionine dependent (HTC) and the other found to be methionine independent (Phi-1). The methionine-dependent cell line (HTC) was discovered to contain markedly less methionine synthase activity, the enzyme activity being less responsive to methionine concentration than in the methionine-independent cells (Phi-1). HTC cells had lower cysteine requirements and contained larger concentrations of reduced glutathione (GSH) and taurine than the Phi-1 cells. Also, in contrast to Phi-1 cells, no glutathione was found in the media of the HTC cells, although large quantities of cysteinylglycine were detected. These results suggested that differences in methionine synthase activity might be partly responsible for methionine dependence and that methionine-dependent cells may have different metabolic requirements for other sulphur amino acids. PMID- 11853691 TI - Selective inhibition of the activities of both eukaryotic DNA polymerases and DNA topoisomerases by elenic acid. AB - (R)-(-)-Elenic acid (R-2,4-dimethyl-22-(p-hydroxyphenyl)-docos-3(E)-enoic acid) (EA) is a DNA topoisomerase II inhibitor found in an Indonesian sponge, Plakinastrella sp. We found and report here that it is a potent inhibitor of calf DNA polymerase alpha (IC(50)=7.7 microM) and rat DNA polymerase beta (IC(50)=12.9 microM). EA did not bind to DNA directly. EA did not influence the activities of DNA polymerases such as plant DNA polymerases I and II and prokaryotic DNA polymerases such as Escherichia coli DNA polymerase I, or other DNA metabolic enzymes such as human immunodeficiency virus (HIV) reverse transcriptase, T7 RNA polymerase and bovine deoxyribonuclease I. Interestingly, EA was also an inhibitor of DNA topoisomerases I and II, although the enzymatic characteristics including modes of action, amino acid sequences and three-dimensional structures were markedly different from those of DNA polymerases. EA could prevent the growth of NUGC-3 cancer cells, and the LD(50) value was 22.5 microM. The cells were halted at G1 and G2/M phase in the cell cycle. From these results, the action mode of EA is discussed. PMID- 11853690 TI - Differential effects of 4-aminoquinoline-containing antimalarial drugs on hemoglobin digestion in Plasmodium falciparum-infected erythrocytes. AB - Several reports suggest that the antimalarial mode of action of quinoline drugs may differ in their mechanistic details. The malaria parasite Plasmodium falciparum was treated in culture with chloroquine, amodiaquine, quinine and mefloquine in a dose- and time-dependent fashion. After removal of the drug, the viability of the parasites and their hemoglobin content were determined. Whereas in the presence of chloroquine and amodiaquine, there was a correlation between parasite killing and accumulation of hemoglobin, with quinine and mefloquine parasite killing was not associated with the accumulation of hemoglobin. Mefloquine inhibited the chloroquine-dependent accumulation of hemoglobin. It is suggested that whereas chloroquine and amodiaquine inhibit the digestion of hemoglobin, mefloquine and possibly quinine inhibit the ingestion of host cell hemoglobin by interfering with the ingestion process. These results may explain the demonstrable antagonism between chloroquine and mefloquine and their antipodal sensitivity to these drugs. PMID- 11853692 TI - UDP-glucuronosyltransferase activity, expression and cellular localization in human placenta at term. AB - The activity, expression and localization of the UDP-glucuronosyltransferases (UGTs) were investigated in human placenta at term. UGT activity (measured with the substrate 4-methylumbelliferone (4-MU)) was observed in all 25 placentas sampled and maximum velocity (V(max)) ranged 13-fold from 5.1+/-0.9 to 66.9+/ 17.5 nmol/min/mg protein (mean+/-SD). Substrate affinity (K(m)) ranged 5-fold from 246+/-24 to 1124+/-422 microM. Using reverse transcriptase-polymerase chain reaction (RT-PCR), expression of the isoforms UGT2B4, 2B7, 2B10, 2B11 and 2B15 was observed in all (12/12) placentas sampled and expression of UGT2B17 was noted in 8/12 placentas. Northern analysis of the UGT2B7 isoform in 12 placentas revealed a 10-fold difference in expression with RT-PCR variability and the 13 fold variation observed in UGT activity. The presence of UGT2B4 and 2B7 proteins (52 and 56kDa, respectively) was demonstrated by Western blotting. The sites of placental UGT2B transcription (in situ hybridization) and protein expression (immunohistochemistry) were located in the syncytium of the placental trophoblasts bordering the placental villi. UGT1A proteins could not be observed with immunohistochemistry or Western blotting and expression could not be observed with RT-PCR. Our discovery of UGT expression and activity at the site of maternal-fetal exchange is consistent with a role for UGTs in detoxification of exogenous and endogenous ligands and the maintenance of placental function through clearance and regulation of steroid hormones. PMID- 11853693 TI - Actions of derivatives of dehydroaltenusin, a new mammalian DNA polymerase alpha specific inhibitor. AB - Dehydroaltenusin was found to be an inhibitor of mammalian DNA polymerase alpha (pol. alpha) in vitro, but did not influence the activities of the other replicative DNA polymerases including even other vertebrate pol. alpha. In this study, we purified or synthesized various slightly modified derivatives of dehydroaltenusin, and using them, investigated the relationship between the chemical structure and the inhibitory effects, and the in vitro and in vivo effects of dehydroaltenusin to determine to what extent the pol. alpha activity inhibition influences cell proliferation. Most of the derivatives lost the enzyme species-specific inhibitory effect, suggesting that dehydroaltenusin is three dimensionally inserted into a pocket present only in mammalian pol. alpha. Dehydroaltenusin inhibited the cell proliferation of the human gastric cancer cell line NUGC-3 by arresting the cells at G1/S-phase, and prevented the incorporation of thymidine into the cells, indicating that it blocks the primary step of in vivo DNA replication by inhibiting pol. alpha. This compound also induced apoptosis of the cells. Dehydroaltenusin is a mammalian pol. alpha specific inhibitor useful in both of in vivo and in vitro experiments. PMID- 11853694 TI - Lack of association between CYP2A5 induction and apoptosis in mouse primary hepatocytes. AB - Upregulation of mouse hepatic cytochrome P450 2A5 (CYP2A5) is a process closely associated with hepatocellular damage and formation of liver tumours. 2 Aminopurine, a protein kinase inhibitor modulating cell cycle control, was recently shown to strongly induce CYP2A5 in mouse hepatocytes. The objective of this study was to determine the association between CYP2A5 induction and apoptosis in mouse primary hepatocytes. Five well-characterised CYP2A5 inducers were tested for their ability to affect apoptosis rate, determined by immunohistochemical in situ 3'-end-labelling technique, in a primary mouse hepatocyte model. Transforming growth factor beta (TGFbeta) was used as a positive (proapoptotic) control. Phenobarbital, pyrazole and the mitogen activated protein kinase inhibitor PD98059 did not significantly affect apoptosis rate in hepatocytes. Norcocaine induced apoptosis at 6 hr (1.8-fold) and 2 aminopurine 12 hr (1.4-fold) after treatment, which is considerably earlier than peaks in the amount of CYP2A5 mRNA. TGFbeta reduced CYP2A5 marker activity, coumarin 7-hydroxylase by 74%. These results indicate that in a primary hepatocyte model (a) there is no systematic correlation between apoptosis and CYP2A5 induction; (b) phenobarbital does not significantly affect the rate of apoptosis; and (c) the induction of apoptosis caused by the chemicals tested occurs considerable earlier than elevation of CYP2A5 expression. Thus, no causal link appears to exist between induction of CYP2A5 and apoptotic rate. PMID- 11853695 TI - In vitro induction of apoptosis vs. necrosis by widely used preservatives: 2 phenoxyethanol, a mixture of isothiazolinones, imidazolidinyl urea and 1,2 pentanediol. AB - Preservatives are added to many final products, such as detergents, cosmetics, pharmaceuticals and vaccines. We conducted an in vitro investigation of the apoptosis- and necrosis-inducing potential of brief applications (10 min) of four common preservatives: ethylene glycol monophenyl ether, 2-phenoxyethanol (EGPE), imidazolidinyl urea (IMU), a mixture of 5-chloro-2-methyl-4-isothiazolin-3-one and 2-methyl-4-isothiazolin-3-one (CMI/MI), and 1,2-pentanediol, a "preservative non-preservative" best known as pentylene glycol. Using HL60 cells, we monitored the kinetics of cell toxicity with the MTT test and analysed extranuclear end points of apoptosis, i.e. phosphatidylserine exposure and nuclear fragmentation. Preservative treatment resulted in a dose-dependent decrease of cell viability. The mode of cell death was dose-dependent: necrosis occurred at high concentrations while apoptosis, shown by DNA laddering, DNA sub-diploid peak and caspase-3 activation, occurred at lower concentrations 0-24hr after exposure to a single dose: CMI/MI induced apoptosis at low concentrations (0.001-0.01%) and necrosis at high concentrations (0.5-0.1%); IMU and EGPE required higher concentrations to induce apoptosis (IMU 0.01-0.1% and EGPE 0.01-0.5%) or necrosis (IMU 0.5-1% and EGPE only at 1%). PG induced apoptosis only at 5%. Externalization of PS, a hallmark of apoptosis, occurred early in HL60 treated with low concentrations of CMI/MI and EGPE and was concomitant with the subdiploid peak in HL60 treated with PG. However, it did not occur in HL60 treated with IMU. In conclusion, at appropriate concentrations, each of the four preservatives modulates the apoptotic machinery by a caspase-dependent mechanism. Thus, apoptosis could be a good parameter to evaluate the cytoxicity of these chemical compounds. PMID- 11853696 TI - Anti-cancer action of 4-iodo-3-nitrobenzamide in combination with buthionine sulfoximine: inactivation of poly(ADP-ribose) polymerase and tumor glycolysis and the appearance of a poly(ADP-ribose) polymerase protease. AB - E-ras 20 tumorigenic malignant cells and CV-1 non-tumorigenic cells were treated with a drug combination of 4-iodo-3-nitrobenzamide (INO(2)BA) and buthionine sulfoximine (BSO). Growth inhibition of E-ras 20 cells by INO(2)BA was augmented 4-fold when cellular GSH content was diminished by BSO, but the growth rate of CV 1 cells was not affected by the drug combination. Analyses of the intracellular fate of the prodrug INO(2)BA revealed that in E-ras 20 cells about 50% of the intracellular reduced drug was covalently protein-bound, and this binding was dependent upon BSO, whereas in CV-1 cells BSO did not influence protein binding. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was identified as the protein that covalently binds the reduction product of INO(2)BA, which is 4-iodo-3 nitrosobenzamide. Since only the enzymatically reduced drug INOBA bound covalently to GAPDH, the BSO-dependent covalent protein-drug association indicated an apparent nitro-reductase activity present in E-ras 20 cells, but not in CV-1 cells, explaining the selective toxicity. Covalent binding of INOBA to GAPDH inactivated this enzyme in vitro; INO(2)BA+BSO also inactivated cellular glycolysis in E-ras 20 cells because it provided the precursor to the inhibitory species: INOBA. Another event that occurred in INO(2)BA+BSO-treated E-ras 20 cells was the progressive appearance of a poly(ADP-ribose) polymerase protease. This enzyme was partially purified and characterized by the polypeptide degradation product generated from PARP I, which exhibited a 50kDa mass. This pattern of proteolysis of PARP I is consistent with a drug-induced necrotic cell killing pathway. PMID- 11853697 TI - Activation of protein kinase C for protection of cells against apoptosis induced by the immunosuppressor prodigiosin. AB - Prodigiosin (PG) is a red pigment produced by Serratia marcescens with immunosuppressive activity. We had recently shown that PG-induced apoptosis in several cancer cell lines including Jurkat-T cells, while acting rapidly, potently and with no marked toxicity in non-malignant cells. Here we examine the role of protein kinase C (PKC) in the regulation of apoptosis triggered by PG. We evaluated the use of phorbol-myristate acetate (PMA) in the inhibition of apoptosis induced by PG in Jurkat-T cells by using FACS analysis of the phosphatidylserine externalisation, Hoechst 33342 staining and fragmentation pattern of DNA as well as proteolysis of poly-(ADP) ribose polymerase (PARP). The anti-apoptotic effect of PMA was accompanied by phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2). Pretreatment of cells with MEK inhibitor PD98059 inhibited PMA-induced phosphorylation of ERK1/2 and the cytoprotective ability of PMA. These results suggest that activation of PKC in Jurkat-T cells confer protection against apoptosis induced by PG and that ERK1/2 mediate anti apoptotic PKC signaling. PMID- 11853698 TI - Modulation of apoptosis by mitochondrial uncouplers: apoptosis-delaying features despite intrinsic cytotoxicity. AB - Disruption of mitochondrial electron transport and opening of the so-called mitochondrial permeability transition pores (PTPs) are early events in apoptotic cell death and may be caused by the uncoupler of mitochondrial oxidation and phosphorylation, carbonyl cyanide p-trifluoromethoxyphenylhydrazone (FCCP). We investigated the cellular toxicity of FCCP in HL60 and CCRF-CEM cells alone or in combination with the known apoptosis inducers such as inhibitor of serine/threonine protein kinases staurosporine (Sts) and protein kinase C inhibitor chelerythrine. FCCP induced apoptotic cell death in both cell lines in a dose-dependent manner, and we were able to demonstrate an appearance of caspase 3-dependent PARP cleavage fragments with Western blot and the appearance of large (15-50 kb) DNA fragments using pulsed-field gel electrophoresis. After 2 hr of incubation with Che or Sts more than half of the cells had died by apoptosis. We observed a statistically significant delay in Sts- and Che-induced apoptotic cell death in CCRF-CEM cells when the cells were preincubated with FCCP but not with zVAD-FMK: about 50% more cells survived after pre-treatment with FCCP, as compared to 1 hr treatment with Che alone (P<0.05), and 25% more cells were alive after 6 hr of treatment, as compared to 6 hr exposure to Sts alone (P<0.05). The protective effect of FCCP was, however, transient and lasted only 6 hr. Treatment with aurintricarboxylic acid completely prevented Che- and Sts-induced apoptotic cell death in CCRF-CEM and HL60 cells. Incubation with Che resulted in a drop in the intracellular ATP content, predominantly distinctive in HL60, and in NAD(+) content in CCRF-CEM cells. Both ATP and NAD(+) drop were prevented with ATA, but not with FCCP or zVAD. Our data suggest that treatment with uncouplers of oxidative phosphorylation can induce apoptotic cell death in haematopoietic cell lines. However, when used in combination with serine/threonine protein kinase inhibitors FCCP can even prevent apoptosis. PMID- 11853699 TI - In vivo nitric oxide transfer of a physiological NO carrier, dinitrosyl dithiolato iron complex, to target complex. AB - Dinitrosyl dithiolato iron complex (DNIC) has been identified as an endogenous NO carrier, yet in vivo mechanisms of NO donation remain undefined. Transnitrosylation, in which a coordinated NO group is transferred to another metal complex, has been observed in transition-metal-nitrosyl chemistry. In this study, we used three kinds of iron dithiocarbamate complexes (Fe-DTCs) as NO acceptors to elucidate in vivo transnitrosylation of diglutathionyl dinitrosyl iron complex [DNIC-(GS)(2)]. Fe-DTCs were administered to mice after the injection of DNIC-(GS)(2) and electron paramagnetic resonance (EPR) spectra were measured both in the resected organs and in the upper abdomen of living mice. The spectral feature gradually changed from an initial DNIC-(GS)(2) signal to mononitrosyl iron dithiocarbamate one, suggesting that NO-Fe-DTC was formed through in vivo reaction of DNIC-(GS)(2) with Fe-DTC. The spectral results in in vitro and in vivo systems indicate that NO-Fe-DTCs can be formed not only by the transfer of coordinated NO-group(s) in DNIC-(GS)(2) but also by the abstraction of Fe-NO group in DNIC-(GS)(2) by free DTC ligands. Transnitrosylation proceeded more rapidly in blood than in liver and kidney; and more efficiently in kidney than in liver. Further, the ability to accept NO from DNIC was dependent on water solubility of Fe-DTCs. Thus, in vivo transnitrosylation from DNIC to exogenous iron complex could be observed and this reaction was influenced by biological constituents and properties of iron complex. These results demonstrate that the transnitrosylation from DNIC to intrinsic NO acceptors like metalloproteins has a probable significance in in vivo NO transfer process. PMID- 11853700 TI - Protective effect of boldine on dopamine-induced membrane permeability transition in brain mitochondria and viability loss in PC12 cells. AB - Boldine ([S]-2,9-dihydroxy-1,10-dimethoxyaporphine) has been shown to exert antioxidant and anti-inflammatory effects. The present study elucidated the protective effect of boldine on catecholamine-induced membrane permeability transition in brain mitochondria and viability loss in PC12 cells. Dopamine (200 microM) and 6-hydroxydopamine (6-OHDA, 100 microM) attenuated Ca(2+) and succinate-induced mitochondrial swelling and membrane potential formation. Boldine (10-100 microM) and 10 microg/mL of superoxide dismutase (SOD) or catalase reduced the effect of catecholamine oxidation on brain mitochondria. Boldine, SOD, and catalase decreased catecholamine-induced mitochondrial cytochrome c release. Antioxidant enzymes attenuated the depressant effect of catecholamines on mitochondrial electron flow, whereas boldine did not reduce it. Boldine inhibited the catecholamine-induced decrease in thioredoxin reductase activity and the increase in thiol oxidation in mitochondria. It also showed a scavenging action on hydrogen peroxide and hydroxyl radicals and decreased the formation of melanin from dopamine. Boldine and antioxidant enzymes decreased the dopamine-induced cell death, including apoptosis, in PC12 cells. The results suggest that boldine may attenuate the catecholamine oxidation-induced brain mitochondrial dysfunction and decrease the dopamine-induced death of PC12 cells through a scavenging action on reactive oxygen species and inhibition of melanin formation and thiol oxidation. PMID- 11853701 TI - Enhancement of phosphatidylcholine biosynthesis by angiotensin-(1-7) in the rat renal cortex. AB - In the present paper, we investigated the effect of angiotensin-(1-7) (Ang-(1-7)) on phospholipid biosynthesis in the rat renal cortex. A significant increase in phosphatidylcholine (PC) labeling was observed when cortical slices, prelabeled with [32P]orthophosphate, were incubated for 30 min in the presence of Ang-(1-7) (1 pM to 100 nM). Neither the phospholipase C inhibitors, neomycin or db-cAMP nor the protein kinase C inhibitors, chelerythrine or H7, modified the stimulatory effect induced by 0.1 nM Ang-(1-7). The enhancement of PC biosynthesis caused by 0.1 nM Ang-(1-7) was unmodified by either losartan, an AT(1) receptor antagonist, or (1-[[4-(dimethylamino)-3-methylphenyl]methyl]-5-(diphenylacetyl)-4,5,6,7 tetrahydro-1H-imidazol[4,5-c]pyridine-6-carboxylic acid ditrifluoroacetate) (PD 123319), an AT(2) receptor antagonist, but was partially blocked by [D-Ala(7)]Ang (1-7), an Ang-(1-7) specific antagonist. However, losartan potentiated the effect of 100 nM Ang-(1-7) on PC biosynthesis. Losartan by itself increased the de novo synthesis of PC. These results suggest that the Ang-(1-7)-mediated increase in PC biosynthesis is independent of AT(1) and AT(2) receptor activation but mediated by a specific Ang-(1-7) receptor. This mechanism is independent of phospholipase C and PKC activation. PMID- 11853702 TI - In vitro and in vivo effects of new insulin releasing agents. AB - The present study aimed at characterizing in vitro and in vivo the effects of BM 208 (N-[4-(5-chloro-2-methoxybenzamidoethyl)benzenesulfonyl]-N'-cyano-N" cyclohexylguanidine) and BM 225 (1-[4-(5-chloro-2-methoxybenzamidoethyl)benzene sulfonamido]-1-cyclohexylamino-2-nitroethylene); two new isosteres of the hypoglycemic sulfonylurea glibenclamide. In rat pancreatic islets perifused at close to normal (8.3mM) D-glucose concentration, both BM 208 and BM 225 (10 and 25 microM) increased 45Ca outflow and insulin release. The compounds did not affect the 45Ca outflow rate from islets exposed to Ca(2+)-free media. In single pancreatic islet cells loaded with the fluorescent Ca(2+) indicator fura-2 and incubated in the presence of 8.3mM glucose, BM 208 and BM 225 raised the [Ca(2+)](i). All these findings indicate that, in islet cells exposed to a physiological concentration of D-glucose, the secretory capacity of the new glibenclamide isosteres is related to a facilitation of Ca(2+) entry. The potency and duration of action of BM 225 was, however, more pronounced than that of BM 208. Successive additions of BM 208 provoked repeated increments in 45Ca outflow and insulin release, without evidence of tachyphylaxis. Lastly, intraperitoneal injection of BM 208 and BM 225 to fed rats lowered plasma glucose concentration in a dose-dependent manner. BM 225 was more potent and acting faster than BM 208. Our results indicate that appropriate structural modification can generate isosteres of glibenclamide with different features and activity profiles. PMID- 11853703 TI - Effects of a selective cyclooxygenase-2 inhibitor, celecoxib, on bone resorption and osteoclastogenesis in vitro. AB - The effects of an important new anti-inflammatory agent, the selective cyclooxygenase-2 inhibitor celecoxib, on bone resorption and osteoclastogenesis elicited by the inflammatory cytokines interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha), the endotoxin lipopolysaccharide (LPS), and the systemic hormones 1alpha,25-dihydroxyvitamin D(3) and parathyroid hormone were examined in vitro. Bone resorption was evaluated by measuring calcium released into the culture medium in a neonatal mouse calvarial bone organ culture. Osteoclastogenesis was evaluated by measuring tartrate-resistant acid phosphatase activity in the cells in cocultures of bone marrow cells and osteoblastic cells and in macrophage-colony-stimulating factor-dependent bone marrow cell cultures. Celecoxib (0.1 microM) completely inhibited the calcium release induced by IL-1beta, TNF-alpha, and LPS. The resorptive effect of 1alpha,25-dihydroxyvitamin D(3) was inhibited partially by celecoxib. In contrast, celecoxib did not inhibit the calcium release elicited by parathyroid hormone or prostaglandin E(2). Celecoxib (0.1 microM) also markedly inhibited osteoclastogenesis induced by these stimulators of bone resorption except for PGE(2) in the coculture system, whereas it failed to inhibit osteoclastogenesis in macrophage-colony-stimulating factor-dependent bone marrow cell cultures. These results indicate that, under certain conditions, cyclooxygenase-2-dependent prostaglandin synthesis is critical for the bone resorption induced by IL-1beta, TNF-alpha, and LPS, and for the osteoclastogenesis induced by these pro inflammatory molecules and calciotropic hormones. The prevention of prostaglandin synthesis by inflammatory cytokines in bone cells could contribute to the efficacy of celecoxib in preventing bone loss in rheumatoid arthritis. PMID- 11853704 TI - Regulatory effects of deoxycholic acid, a component of the anti-inflammatory traditional Chinese medicine Niuhuang, on human leukocyte response to chemoattractants. AB - Niuhuang is a commonly used Chinese traditional medicine with immunoregulatory and anti-inflammatory properties. Deoxycholic acid (DCA) is a major active constituent of Niuhuang. The reaction of human leukocytes to chemoattractants is an important part of the host immune response and also plays a crucial role in the development of inflammation. We, therefore, investigated the in vitro effects of DCA on human monocyte and neutrophil responses to classic chemoattractants [fMet-Leu-Phe (fMLP), complement fraction 5a (C5a)], CC chemokine [monocyte chemoattractant protein-1 (MCP-1/CCL2)], and/or CXC chemokines [stromal cell derived factor-1 (SDF-1alpha/CXCL12), interleukin-8 (IL-8/CXCL8)]. The results showed that DCA significantly inhibited fMLP-induced monocyte and neutrophil chemotaxis and calcium mobilization, and also blocked the binding of [3H]fMLP and anti-formyl peptide receptor (FPR) monoclonal antibodies (mAb) to the cells. The inhibitory effects of DCA on calcium mobilization and anti-FPR-mAb binding to the receptor could be abrogated by washing DCA out of the cell suspension, suggesting that DCA blocked fMLP receptors via a steric hindrance mechanism, not via receptor internalization. DCA had no significant inhibitory effects on MCP-1-, SDF-1alpha-, or C5a-induced monocyte function, or C5a- or IL-8-induced neutrophil function. Taken together, our experimental results suggest that blockade of fMLP receptors may contribute to the anti-inflammatory effects of traditional medicine containing DCA. PMID- 11853705 TI - Mitochondrial gene expression and ventricular fibrillation in ischemic/reperfused nondiabetic and diabetic myocardium. AB - We investigated the mitochondrial gene expression related to cardiac function and ventricular fibrillation (VF) in ischemic/reperfused nondiabetic and diabetic myocardium. To identify potentially more specific gene responses we performed subtractive screening, Northern blotting, and reverse transcription-polymerase chain reaction (RT-PCR) of mitochondrial genes expressed after 30 min ischemia followed by 120 min reperfusion in isolated rat hearts that showed VF or did not show VF. Cytochrome oxidase B subunit III (COXBIII) and ATP synthase subunit 6, studied and selected out of 40 mitochondrial genes by subtractive screening, showed an expression after 30 min ischemia (no VF was recorded) in both nondiabetic and diabetic subjects. Upon reperfusion, the down-regulation of these genes was only observed in fibrillated hearts. Such a reduction in signal intensity was not seen in nonfibrillated myocardium. In additional studies, nondiabetic and diabetic hearts, without the ischemia/reperfusion protocol, were subjected to electrical fibrillation, and a significant reduction in COXBIII and ATPS6 mRNA signal intensity was observed indicating that VF contributes to the down-regulation of these genes. Cardiac function (heart rate, coronary flow, aortic flow, left ventricular developed pressure) showed no correlation between the up- and down-regulation of these mitochondrial genes in both nondiabetic and diabetic ischemic/reperfused myocardium. Our data suggest that COXBIII and ATPS6 may play a critical role in arrhythmogenesis, and the stimulation of COXBIII and ATPS6 mRNA expression may prevent the development of VF in both nondiabetic and diabetic ischemic/reperfused myocardium. PMID- 11853706 TI - Effect of malondialdehyde-acetaldehyde-protein adducts on the protein kinase C dependent secretion of urokinase-type plasminogen activator in hepatic stellate cells. AB - Previous studies from our laboratory have shown that malondialdehyde-acetaldehyde protein adducts (MAA adducts) are formed in hepatocytes of ethanol-fed rats and directly influence the hepatic stellate cells (HSCs) to induce their secretion of chemokines and to up-regulate their expression of adhesion molecules. Since protein kinase C (PKC) is known to play a major role in many diverse intracellular signal transduction processes, we investigated whether MAA adducts influence the function of HSCs via a PKC-dependent pathway. HSCs in culture were exposed to MAA adducts, and PKC activity was determined. We observed a time- and concentration-dependent activation of PKC when cultures were exposed to BSA-MAA as compared with unmodified BSA. Using PKC isoform-specific inhibitors, we also showed that BSA-MAA induces the activation of a specific isoform of PKC, PKC alpha, in HSCs. No activation of PKC was observed when HSCs were exposed to other aldehyde adducts such as BSA-acetaldehyde or BSA-malondialdehyde, indicating that the effects of MAA adducts on HSCs were somewhat specific. We further examined whether the observed increase in PKC activation induced by MAA adducts in HSCs, in turn, causes a functional effect. We observed that BSA-MAA induces the increased secretion of urokinase-type plasminogen activator, a key component of the plasmin-generating system, and that PKC activation is necessary for this enhanced urokinase-type plasminogen activator secretion. These results indicate that MAA adducts via a PKC-mediated pathway may regulate plasmin-mediated matrix degradation in the liver, thereby contributing to the progression of hepatic fibrosis. PMID- 11853707 TI - Autoantibody against neuron-specific enolase found in glaucoma patients causes retinal dysfunction in vivo. AB - PURPOSE: In our recent paper, we have reported the presence of serum autoantibody against neuron-specific enolase (NSE) in patients with glaucoma. The purpose of the present study was to investigate further the pathological effects of anti-NSE antibody on retina by comparing them with the effects induced by N-methyl-D aspartate (NMDA). METHODS: Either a glaucoma patient's serum or purified anti-NSE antibody, or 10-40 mM NMDA was intravitreously administered into Lewis rat eyes, and electrophysiological, histopathological, and biochemical evaluations were performed. In addition, the neuroprotective effects of anti-glaucoma drugs, such as timolol, betaxolol, nipradilol, and isopropyl unoprostone, and a calcium antagonist were also studied using these animal models. RESULTS: Electron microscopy revealed that intravitreal administration of a glaucoma patient's serum, which immunoreacted with retinal 50 kDa in Western blot analysis, and purified anti-NSE antibody induced retinal ganglion cell apoptosis in rat eyes. Functionally, these eyes showed a significant decrease in electroretinogram (ERG) responses and a remarkable decrease in rhodopsin phosphorylation reaction. These changes were comparable to the effects observed after the intravitreal administration of 20 mM NMDA. Co-administration of nipradilol, an alpha- and beta blocker, with anti-NSE antibody or 20 mM NMDA caused marked recovery of the affected ERG responses within 2 weeks. In contrast, administration of timolol or betaxolol showed no recovery effect on the ERG responses. Among these drugs, only betaxolol showed a recovery effect on NMDA-induced decrease of rhodopsin phosphorylation. Nilvadipine functioned beneficially on both impaired ERG and rhodopsin phosphorylation reactions observed in rat eyes injected intravitreously with anti-NSE antibody or NMDA. These effects of nilvadipine were not changed by the addition of endothelin-1. In contrast, isopropyl unoprostone had no effect on these functions. CONCLUSION: These observations suggest that serum autoantibody against NSE found in some patients with glaucoma induces retinal dysfunction in vivo, similarly to NMDA. PMID- 11853708 TI - Clinical significance of serum antibody against neuron-specific enolase in glaucoma patients. AB - PURPOSE: In a recent study, we found the presence of serum autoantibody against neuron-specific enolase (NSE) in glaucoma patients. The purpose of the present paper is to investigate further the clinical significance of the presence of the serum antibody against NSE in glaucoma patients. METHODS: Serum autoantibody against NSE was examined by Western blot analysis in 143 patients with glaucoma (normal tension glaucoma [NTG], 45 cases; primary open angle glaucoma [POAG] 98 cases). Clinical characteristics including visual acuity, visual field, intraocular pressure (IOP), and optic disc features were compared between the serum autoantibody-positive and the serum autoantibody-negative patients. RESULTS: Maximum IOP in the serum anti-NSE antibody-positive patients was significantly lower than that in the negative patients (P <.05). However, no statistical differences were observed in visual field loss, disc cupping, or other clinical factors. During the clinical course, rates of the presence of anti NSE antibody were significantly higher in the early stages of POAG (P <.0001) with visual field deterioration than without it. Although it was not statistically significant, the positive rates of serum anti-NSE antibody were relatively higher in the later stages of POAG and NTG with visual field deterioration than without it. CONCLUSION: The present observations suggest that the presence of serum autoantibody against NSE may be clinically useful for predicting the progression of visual field loss in POAG patients. PMID- 11853709 TI - Elevated soluble Fas in aqueous humor of patients with Behcet's uveitis: correlation with uveitis severity. AB - PURPOSE: To examine the role of soluble Fas (sFas) in patients with Behcet's uveitis. METHODS: We measured the sFas levels in both sera and aqueous humor (AH) of patients (n = 40) with uveitis and of non-uveitis controls (n = 27) using an enzyme-linked immunosorbent assay. The patients with uveitis comprised 24 with Behcet's disease, 6 pan-uveitis, 5 anterior uveitis, 2 lens-induced uveitis, 1 Vogt-Koyanagi-Harada disease, 1 sarcoidosis, and 1 retinal vasculitis. The severity of uveitis was determined by the Hogan grading method (0--4 grade) at the time of sampling. RESULTS: The concentration of aqueous sFas in uveitis patients was significantly higher than that in non-uveitis controls, while there was no difference in the serum concentration of sFas between the two groups. In the paired samples of serum and AH, obtained simultaneously, the aqueous sFas levels were higher than serum Fas levels in patients with uveitis, whereas the non-uveitis controls displayed significantly lower sFas levels in AH than in the serum. The sFas levels in AH or serum were not different between Behcet's uveitis patients and non-Behcet's uveitis patients. However, in patients with Behcet's uveitis, circulating sFas strongly correlated with aqueous sFas, which was not so in those with non-Behcet's uveitis. Patients (n = 29) with more active (grade greater-than-or-equal 2) uveitis had significantly higher levels of aqueous sFas than those (n = 11) with less active (grade < 2) uveitis. After treatment with steroids and/or immunosuppressive agents, aqueous sFas levels decreased in parallel with a reduction in the number of inflammatory cells. CONCLUSIONS: The levels of sFas were elevated in patients with Behcet's uveitis and correlated well with the uveitis severity in these patients. PMID- 11853710 TI - Involvement of mechanical stretch in the gelatinolytic activity of the fibrous sclera of chicks, in vitro. AB - PURPOSE: To investigate the role of mechanical stretch in the regulation of matrix metalloproteinase-2 (MMP-2) in scleral fibroblasts of chick embryos. METHODS: Scleral fibroblasts derived from chick embryos were seeded onto flexible bottom culture plates, and subjected to a pulsatile stretch when the cells became subconfluent. After stretching, the cells and the conditioned medium were harvested. One portion of the conditioned medium was activated by reduction, alkylation, and 4-aminophenylmercuric acetate (APMA) treatment. The conditioned medium, with and without the treatment, was subjected to gelatin zymography and quantitative assays. Total cytoplasmic RNA was extracted from the cells, and the expression of MMP-2 and the tissue inhibitor of metalloproteinase-2 (TIMP-2) mRNA was examined by Northern blot analysis. RESULTS: The predominant gelatinolytic enzyme secreted by scleral fibroblasts was MMP-2. The mechanical stretch increased the gelatinolytic activities significantly in the conditioned medium with reduction, alkylation, and APMA treatment. Mechanical stretch also enhanced the expression of MMP-2 and TIMP-2 mRNA in scleral fibroblasts significantly. CONCLUSIONS: These results suggest that mechanical stretch may be involved in the regulation of the extracellular matrix in the fibrous sclera of chicks in vivo. PMID- 11853711 TI - Long-term effect of topically applied isopropyl unoprostone on microcirculation in the human ocular fundus. AB - PURPOSE: To investigate the long-term effect of 0.12% isopropyl unoprostone (Rescula) on microcirculation in the human ocular fundus. METHODS: A laser speckle tissue circulation analyzer was used to measure normalized blur (NB), a quantitative index of blood flow velocity, in the optic nerve head (ONH) and choroid-retina before and 4.5 hours after the instillation of a placebo into both eyes of 11 healthy volunteers. The intraocular pressure (IOP), blood pressure, and pulse rate were also recorded in this control experiment. Thereafter, a drop of unoprostone or a placebo was instilled into each eye in a double-blind manner twice a day for 21 days to form treated and untreated groups. RESULTS: After 21 days, the NB values in the ONH and choroid-retina had increased significantly and the IOP had decreased significantly in the unoprostone-treated eyes. Ocular perfusion pressure showed no significant change. CONCLUSIONS: These results suggest that long-term application of unoprostone can increase microcirculatory blood flow in the human ocular fundus, probably due to a reduction in vascular resistance. PMID- 11853713 TI - Leber's hereditary optic neuropathy--the spectrum of mitochondrial DNA mutations in Chinese patients. AB - PURPOSE: To investigate the spectrum of mitochondrial DNA (mtDNA) mutations in Chinese patients with Leber's hereditary optic neuropathy (LHON), optic atrophy of unknown etiology, and optic neuropathy of known etiology. METHODS: Twenty seven patients from 25 LHON pedigrees, 22 patients with bilateral optic atrophy of unknown etiology, 21 patients with optic neuropathy of known etiology, and 25 normal healthy controls were included in this study. Twelve pairs of primers that covered the 21 reported mtDNA mutations were utilized. Single-strand conformation polymorphism analysis and DNA sequencing were used to detect base substitutions in mtDNA. RESULTS: Twenty-three LHON pedigrees (92%) had the 11778 mtDNA primary mutation. Two pedigrees (8%) had the 14484 mutation. No 3460 mutations were detected in this group. Thirteen other sequence changes were found in these patients, but only the 4216 mutation had been reported previously. Thirteen pedigrees had multi-mutation patterns consisting of one primary mutation together with other sequence changes. No primary mutations were found in patients with optic atrophy of unknown etiology or in patients with optic neuropathy of known etiology. CONCLUSIONS: High frequency of 11778 mtDNA mutation was found in Chinese patients with LHON. No specific multi-mutation pattern such as the European mtDNA haplogroup J was found. PMID- 11853712 TI - Xenotransplantation of retinal pigment epithelial cells into RCS rats. AB - PURPOSE: Successful engraftment of retinal pigment epithelial cells (RPE) to treat RPE-related retinopathy will depend, at least in part, on controlling the immune response. In order to understand this process we evaluated the fate of RPE xenografts in the subretinal space, anterior chamber, and subcutis of nonimmunosuppressed Royal College of Surgeons rats. METHODS: Freshly isolated adult porcine RPE cells were used as xenografts and implanted when recipients were 17 to 21 days old. The extent of photoreceptor rescue by subretinal transplants was determined by counting the maximum layers of surviving photoreceptor nuclei in histologic sections. Cellular immune response was evaluated by immunohistochemistry. RESULTS: Compared to non- or sham-injected eyes, subretinal xenografts in RPE-transplanted eyes were able to induce a dramatic rescue effect (P <.01). However, the effect was not absolute and photoreceptor cell degeneration was only delayed. Xenografts both in the anterior chamber and in the subcutaneous tissue led to an inflammatory cellular infiltration. CONCLUSION: RPE xenografts in subcutaneous space and in the anterior chamber are rejected by a delayed but vigorous inflammatory cell infiltration. Subretinal RPE xenografts are protected from a strong cellular rejection, but seem to undergo a slow functional deterioration, reflected by a decline in their capability to rescue adjacent photoreceptors. PMID- 11853714 TI - Clinical and sensorial characteristics of microtropia. AB - PURPOSE: To investigate the clinical characteristics and sensory mechanism of microtropia. METHODS: Twenty patients with primary microtropia were studied. The patients were evaluated by the visuscope, Irvine prism test, Bagolini striated glasses test, Worth 4-dot test (W4D), TNO, and stereo-fly plate of the Titmus test. RESULTS: The patients who had positive cover test and eccentric fixation showed wandering eccentric fixation. The Irvine prism test was positive in all the patients by using prisms of different strengths and positions. All the patients had abnormal fusion at near. At distance, 15 patients had abnormal fusion by the Bagolini glasses and 8 patients had abnormal fusion by the Worth test. Ten patients had gross stereopsis and the remaining 10 had no measurable stereopsis. CONCLUSIONS: The type of microtropia with eccentric fixation without identity is mainly due to the wandering eccentric fixation. All patients with microtropia have abnormal fusion without fixation point scotoma because positive prism test response does not change to normal by using prisms of different strengths and positions. Differences in fusion results are mainly due to the weakness of abnormal fusion rather than fixation point scotoma. With occlusion treatment, amblyopia can be improved, whereas binocular defects of microtropia cannot be improved. PMID- 11853715 TI - Concordance of strabismic phenotypes in monozygotic versus multizygotic twins and other multiple births. AB - PURPOSE: The concordance of strabismic phenotypes was examined in monozygotic versus multizygotic twins and other multiple births to study the role of genetic background in different types of commitant strabismus. METHODS: Medical charts of 45 consecutive pairs of twins (16 monozygotic and 18 dizygotic twins, and 11 with unknown zygosity), 3 sets of triplets (one monozygotic and 2 trizygotic triplets), and one set of quadruzygotic quadruplets examined at 6 institutions between 1973 and 1999 were reviewed retrospectively. The concordance was defined as both or all members having either esotropia or exotropia in common. RESULTS: The concordance of strabismic phenotypes was noted in 33 of 49 pairs or sets (67.3%): 14 of 17 monozygotic twins or triplets (82.4%), 10 of 21 multizygotic twins, triplets, or quadruplets (47.6%), and 9 of 11 twins with unknown zygosity (81.8%). The concordance rate was significantly higher in monozygosity than in multizygosity (P =.043, Fisher exact probability test). The predominant concordant phenotypes in monozygosity were accommodative esotropia and intermittent exotropia. CONCLUSION: A high concordance rate of strabismic phenotypes, predominantly of accommodative esotropia and intermittent exotropia in monozygosity, suggests the genetic background for these types of strabismus. PMID- 11853716 TI - The corneal endothelium and thickness in type II diabetes mellitus. AB - PURPOSE: To compare the endothelial structure and thickness of the cornea in diabetic and nondiabetic patients, and to evaluate the systemic and ocular factors that contribute to the damage of endothelial cells in diabetic patients. METHODS: The corneal endothelial structure and central corneal thickness (CCT) were investigated in 99 type II diabetic patients (99 eyes) and in 97 nondiabetic patients (97 eyes). The endothelial structure was examined for cell density, coefficient of variation of cell area, and percentage of hexagonal cells. The correlation between CCT and the grade of diabetic retinopathy was evaluated. Multivariate regression analysis was performed to assess systemic factors (patient age, sex, duration of diabetes mellitus, hemoglobin A(1c) value, glucose in urine, blood urea nitrogen value, and creatine value) and ocular factors (grade of diabetic retinopathy and history of photocoagulation) related to endothelial cell density. RESULTS: The endothelial cell density was decreased and the coefficient of variation of cell area was increased in diabetic patients (P <.05). However, the percentage of hexagonal cells and CCT in diabetic patients was not significantly different from that in nondiabetic patients. CCT was similar regardless of the stage of diabetic retinopathy. Multivariate regression analysis indicated that none of the systemic or ocular factors was significantly correlated with the endothelial cell density. CONCLUSIONS: Corneal endothelial cell structure was damaged, but CCT was not increased in type II diabetic patients. There were no systemic or ocular factors at any one point to induce corneal endothelial damage. PMID- 11853717 TI - Varicella-zoster viral antigen identified in iridocyclitis patient. AB - BACKGROUND: The varicella-zoster virus (VZV) antigen has not been identified immunohistologically in iridocyclitis due to VZV. CASE: A 65-year-old woman diagnosed with iridocyclitis and secondary glaucoma underwent trabeculectomy. Samples of aqueous humor and juxtacanalicular and iris tissue were obtained for immunohistological and polymerase chain reaction (PCR) study. OBSERVATIONS: Slit lamp microscopy revealed ciliary injection, corneal epithelial edema, mutton fat precipitates, flare, cells, and progressive iris atrophy in the right eye. Subsequently, scant eruptions on her right upper eyelid appeared and disappeared within a week. Although a diagnostic increase in the complement fixation antibody titer to VZV was not observed, we started medical treatment for VZV, on suspicion of iridocyclitis due to VZV. Despite medical treatment, the ratio of peripheral anterior synechia was greater than 60% and iris atrophy progressed in parallel. The intraocular pressure in the right eye remained above 30 mm Hg at 6 months after the first visit, so trabeculectomy was performed. VZV-specific DNA was detected in the aqueous humor by the PCR study. Immunohistological examination demonstrated numerous VZV antigen-positive cells in the iris stroma, in particular, vascular endothelial cells. CONCLUSION: To our knowledge, this is the first report of the detection of VZV antigen in the iris of an iridocyclitis patient. PMID- 11853718 TI - A case of acute angle-closure glaucoma secondary to posterior scleritis in patient with Sturge-Weber syndrome. AB - BACKGROUND: Sturge-Weber syndrome has been known to be frequently associated with facial cutaneous angioma and ipsilateral glaucoma. However, as far as we know, no cases accompanied by acute angle-closure glaucoma have been reported in patients with Sturge-Weber syndrome. CASE: A 14-year-old boy with unilateral acute angle closure glaucoma secondary to posterior scleritis associated with Sturge-Weber syndrome is described. OBSERVATIONS: Slit-lamp examination revealed diffuse episcleral venous hemangioma in the right eye. With ultrasound biomicroscopy, a forward shift of the lens-iris diaphragm, a swelling of the ciliary body, and an anterior rotation of the ciliary processes with annular choroidal effusion were detected. The patient responded well to treatment with systemic corticosteroids and cycloplegics. CONCLUSIONS: In our patient, inflammatory changes of the sclera, including swelling of the ciliary body, choroidal effusion, an anterior rotation of the ciliary processes at the scleral spur, and swelling of the lens, leading to closure of the anterior chamber angle, were suggested to be the major mechanisms of intraocular pressure elevation. PMID- 11853719 TI - Plasma level of interleukin-6 is an indicator for predicting diabetic macular edema. AB - PURPOSE: To find a predictor in the pathogenesis of macular edema, we investigated ocular and systemic risk factors. METHODS: One hundred and fifty nine patients with mild diabetic retinopathy who showed one or more soft exudates were recruited. We selected the possible predictors on the basis of relevant factors, including concentration of vascular endothelial growth factor, interleukin-6 (IL-6), transforming growth factor (TGF)-beta(1), tumor necrosis factor (TNF)-alpha, and lipoprotein(a) in plasma, and serum level of von Willebrand factor and thrombomodulin. RESULTS: Macular edema was not detected in 94 eyes; focal macular edema was detected in 46 eyes; diffuse macular edema was detected in 18 eyes; and cystoid macular edema was present in 1 eye. The plasma level of IL-6 concentration and the state of the posterior vitreous detachment (PVD) correlated significantly with the severity of macular edema (odds ratios = 3.68, 1.70, respectively). Other risk factors were not significantly associated with macular edema. We estimated the probability of macular edema according to the IL-6 level in plasma and the state of the PVD, and were able to predict the probability of macular edema. CONCLUSION: The results of the present study indicate that IL-6 concentration in plasma and the state of the PVD can be predictors of macular edema. PMID- 11853720 TI - Shortening the duration of prone positioning after macular hole surgery- comparison between 1-week and 1-day prone positioning. AB - PURPOSE: To shorten the duration of prone positioning after macular hole surgery from 1 week to 1 day, and to evaluate preoperative factors and the initial hole closure rate. METHODS: The subjects were 33 patients (34 eyes) who underwent macular hole surgery between April 1998 and August 1999, and maintained the prone position for 1 week (1-week group) and 21 patients (21 eyes) who underwent this operation between September 1999 and March 2000, and maintained the prone position for only 1 day (1-day group). The criteria for macular hole surgery were no more than 6 months since symptom development and no flattening of the fluid cuff. Eyes treated by removal of the retinal pigment epithelium (RPE) or internal limiting membrane (ILM) were excluded. In all phakic eyes, cataract surgery was combined with macular hole surgery. In the 1-day group, the patients maintained the prone position for 1 day and were instructed to avoid only the supine position for the subsequent 1 week. RESULTS: Preoperative factors possibly affecting the hole closure rate did not differ significantly between the two groups. Initial hole closure rates were similar in the 1-week (91.2%) and 1-day (90.5%) groups. CONCLUSION: In eyes without symptoms of long duration, the duration of prone positioning after macular hole surgery may be shortened to 1 day even without using special techniques, such as removal of the RPE or ILM, in combination with macular hole surgery. PMID- 11853721 TI - Correlation of blue chromatic macular sensitivity with optic disc change in early glaucoma patients. AB - PURPOSE: To investigate the relationship between morphological changes of the optic nerve head (ONH) and macular sensitivity determined with blue on yellow (B on Y) and white on white (W on W) perimetry in normal subjects and patients with glaucoma. METHODS: One randomly chosen eye was evaluated in each of 28 healthy subjects, 23 patients with ocular hypertension (OH), and 23 patients with early primary open-angle glaucoma (POAG). The mean macular sensitivity determined with B on Y and W on W perimetry was obtained by a macular program using a modified Humphrey field analyzer. The Heidelberg retina tomograph with software version 2.01 was used to evaluate the temporal topographic measurements of ONH. RESULTS: There was no significant difference in temporal ONH parameters among the three clinical groups. The mean macular sensitivity of B on Y and W on W perimetry in early POAG patients was significantly lower than that in healthy subjects and in patients with OH. The mean macular sensitivity of W on W perimetry showed no significant correlation with temporal ONH parameters in any clinical groups. In patients with early POAG, the mean macular sensitivity of B on Y perimetry was significantly related to cup area and volume, cup/disc area ratio, and rim volume in the temporal sector of the ONH. CONCLUSION: The measurement of the mean macular sensitivity of B on Y perimetry might make it possible to detect functional damage prior to morphological changes in the ONH due to elevated intraocular pressure in glaucoma. PMID- 11853723 TI - Optic nerve involvement in neuro-Behcet's disease. AB - BACKGROUND: To report the ocular manifestations of neuro-Behcet's disease. CASES: A 34-year-old woman had recurrent meningitis. She developed diplopia, headache, and ataxia, and was diagnosed as having neuro-Behcet's disease. OBSERVATIONS: Imaging revealed two infarct foci in the transitional midbrain and pons. After treatment with prednisolone and colchicine, the diplopia resolved. Two years later, a scotoma developed in the right eye, in which the best-corrected visual acuity was 4/200. Papillitis and a prepapillary vitreous opacity were seen in the right fundus. These findings disappeared 11 days after subconjunctival steroid injections and increased colchicine. Her vision gradually improved to 20/20 two months later. CONCLUSION: Neuro-Behcet's disease may manifest with transient optic neuritis and prepapillary vitreous opacity. PMID- 11853722 TI - Peripapillary subretinal neovascularization in sarcoidosis: remission and exacerbation during oral corticosteroid therapy. AB - BACKGROUND: In sarcoidosis, peripapillary subretinal neovascularization is rare. The role of corticosteroid therapy for subretinal neovascularization is controversial. CASE: A 38-year-old female patient weighing 38 kg with histologically diagnosed sarcoidosis presented with peripapillary subretinal neovascularization, retinal phlebitis, a hyperemic disc, and snowball vitreous opacities in the left eye. OBSERVATION: Oral betamethasone therapy at an initial dose of 3 mg/day reduced the size of subretinal neovascular membrane, and the membrane became fibrous. Despite the total initial 140 mg of betamethasone given over 2.5 months and the additional total 700 mg of prednisolone given over the next 2 months, the subretinal neovascularization recurred. Six months after the first recurrence, a second recurrence developed during the tapering-off period of oral corticosteroid therapy. At the second recurrence, the oral corticosteroid therapy was ineffective in reducing the size of the neovascular membrane. CONCLUSION: In our patient, oral corticosteroids temporarily suppressed peripapillary subretinal neovascularization but failed to prevent extension of neovascular membrane to the fovea because of recurrent sarcoidosis. Over time, oral corticosteroids appear to lose their effectiveness for treating repeated recurrence of peripapillary subretinal neovascularization associated with sarcoidosis. PMID- 11853724 TI - Complications in motility PEG placement for hydroxyapatite orbital implant in anophthalmic socket. AB - PURPOSE: In a retrospective study, we evaluated the complications in using the motility peg system (nonsleeved and sleeved) for hydroxyapatite orbital implants in an anophthalmic socket. METHODS: Drilling for motility peg placement was performed in 265 patients with hydroxyapatite implantation: nonsleeved peg system (n = 191), sleeved peg system (n = 74). A statistical analysis was performed using the chi-square test. RESULTS: Extrusion rates were significantly lower in the sleeved peg system (10.8%) compared to the nonsleeved peg system (27.2%) (P =.005). The other complications related to motility peg placement were granulation tissue overgrowth (4.2%), hydroxyapatite exposure around peg head (3.0%), and decentered peg (1.9%). CONCLUSIONS: To minimize peg extrusion, the sleeved peg was better than the nonsleeved peg for use in primary motility peg placement. PMID- 11853725 TI - Visual evoked potentials elicited by pseudorandom stimulation from patients with macular degeneration. AB - PURPOSE: To investigate the effect of a central scotoma on the amplitude, implicit time (IT), and temporal frequency characteristics (TFC) of the visual evoked potentials (VEPs) elicited by a pseudorandom binary sequence (PRBS) stimulus in age-related macular degeneration (AMD) patients. METHODS: Twenty-six patients with AMD, 17 eyes with visual acuity of less than 20/100, and 9 eyes with visual acuity between 20/70 and 20/25, were examined. Nine eyes of age matched healthy volunteers served as controls. To elicit the PRBS-VEPs, one eye was stimulated with a PRBS stimulus. The first-order kernel was calculated from a cross-correlation between the PRBS and the VEPs. The Fourier transformed first order kernel was used as the TFC of the visual system. RESULTS: The mean IT of P2 (second positive peak) of the first-order kernel was significantly delayed (t test, P <.05), and the P2-N2 (peak-to-peak of P2 and second negative peak N2) amplitude was significantly reduced (t-test, P <.01) in eyes with AMD. A depression of the TFC values in the 6-18 Hz band was prominent in patients with AMD (t-test, P <.01). CONCLUSION: PRBS-VEPs demonstrated a prolonged IT and reduced amplitude of the first-order kernel, and reduced TFC with a reduction of visual acuity in patients with macular degeneration. PMID- 11853726 TI - A unique case of primary corneal melanoma. AB - PURPOSE: To report a unique case of primary corneal melanoma. METHODS: A 59-year old white woman underwent keratectomy to remove a pigmented rapidly enlarging growth on the corner of her left eye. The specimen was submitted for histopathologic examination. RESULTS: Ophthalmic examination disclosed no association with limbal or conjunctival melanocytic abnormalities. Histopathologic studies demonstrated nodular malignant melanoma. CONCLUSION: Possible development of primary corneal melanoma should be considered in diagnosing all cases with a past history of corneal melanin pigmentation. PMID- 11853741 TI - Proline metabolism and NAD kinase activity in greenbean plants subjected to cold shock. AB - The aim of the present work was to examine the relationship between proline metabolism and NAD kinase activity in greenbeans submitted to cold-shock. For this, 15-day-old greenbean plants were subjected to a temperature of 4 degrees C (cold shock) for 180 min. Our results indicate that the plants showed foliar accumulation of proline, with the enzymes ornithine-delta-aminotransferase (OAT) and proline dehydrogenase (PDH) appearing as determinant in this accumulation under cold-shock. Also, we found a close relationship between the Ca(2+)-CaM dependent NAD kinase activity and proline metabolism, suggesting that the adaptive responses or acclimation of plants to cold stress are preceded by increased [Ca(2+)](cyt). PMID- 11853742 TI - Microbial transformation of cadina-4,10(15)-dien-3-one, aromadendr-1(10)-en-9-one and methyl ursolate by Mucor plumbeus ATCC 4740. AB - The sesquiterpenes cadina-4,10(15)-dien-3-one (1) and aromadendr-1(10)-en-9-one (squamulosone) (14) along with the triterpenoid methyl ursolate (21) were incubated with the fungus Mucor plumbeus ATCC 4740. Substrates 1, 14 and ursolic acid (20) were isolated from the plant Hyptis verticillata in large quantities. M. plumbeus hydroxylated 1 at C-12 and C-14. When the iron content of the medium was reduced, however, hydroxylation at these positions was also accompanied by epoxidation of the exocyclic double bond. In total nine new oxygenated cadinanes have been obtained. Sesquiterpene 14 was converted to the novel 2alpha,13 dihydroxy derivative along with four other metabolites. Methyl ursolate (21) was transformed to a new compound, methyl 3beta,7beta,21beta-trihydroxyursa-9(11),12 dien-28-oate (22). Two other triterpenoids, 3beta,28-dihydroxyurs-12-ene (uvaol) (23) and 3beta,28-bis(dimethylcarbamoxy)urs-12-ene (24) were not transformed by the micro-organism, however. PMID- 11853743 TI - Biotransformation of cadina-4,10(15)-dien-3-one and 3alpha-hydroxycadina-4,10(15) diene by Curvularia lunata ATCC 12017. AB - Cadina-4,10(15)-dien-3-one (1) was metabolised by Curvularia lunata ATCC 12017 in two different growth media to give three metabolites, one of which, 12 hydroxycadina-4,10(15)-dien-3-one (4), was new. Incubation of 3alpha hydroxycadina-4,10(15)-diene (2) with the fungus produced three new analogues, namely, (4S)-1alpha,3alpha-dihydroxycadin-10(15)-ene (5), 3alpha,14 dihydroxycadina-4,10(15)-diene (6) and 3alpha,12-dihydroxycadina-4,10(15)-diene (7). PMID- 11853744 TI - Effects of Na(2)SO(3) on the activities of antioxidant enzymes in geranium seedlings. AB - This study investigated the effects of Na(2)SO(3), which releases SO(2) in apoplastic water, on the growth of geranium seedlings and on the activities of various antioxidant enzymes including peroxidase. Sodium sulfite (Na(2)SO(3)) addition both inhibited primary root growth and stimulated lateral root growth of the seedlings respectively. In addition, the contents of chlorophyll and Rubisco protein of the seedlings were greatly reduced with Na(2)SO(3) treatment. Total peroxidase activities of the seedlings also increased proportionally with the amount of Na(2)SO(3), this presumably correlating with oxidative stress levels. Notably, about an 8-fold enhancement of total peroxidase activity occurred in seedlings treated with 60 nM Na(2)SO(3) at pH 4.0. This enhancement of total peroxidase activity was mainly due to the increase of a strong cationic isoperoxidase, strong anionic isoperoxidase and neutral isoperoxidase activities. The strong cationic isoperoxidase from geranium seedlings was found to be the same enzyme as PC3 from geranium callus in terms of its physicochemical and catalytic properties. Moreover, the activities of superoxide dismutase and glutathione reductase were greatly enhanced with Na(2)SO(3) treatments at pH 4.0 without significant alteration of catalase activity. These results suggest that Na(2)SO(3) exposure, activities the plants defense mechanism against the reactive oxygen species generated. PMID- 11853745 TI - Cyanohydrin glycosides of Passiflora: distribution pattern, a saturated cyclopentane derivative from P. guatemalensis, and formation of pseudocyanogenic alpha-hydroxyamides as isolation artefacts. AB - Nineteen species of Passiflora (Passifloraceae) were examined for the presence of cyanogenic glycosides. Passibiflorin, a bisglycoside containing the 6-deoxy-beta D-gulopyranosyl residue, was isolated from P. apetala, P. biflora, P. cuneata, P. indecora, P. murucuja and P. perfoliata. In some cases this glycoside co-occurs with simple beta-D-glucopyranosides: tetraphyllin A, deidaclin, tetraphyllin B, volkenin, epivolkenin and taraktophyllin. P. citrina contains passicapsin, a rare glycoside with the 2,6-dideoxy-beta-D-xylo-hexopyranosyl moiety, while P. herbertiana contains tetraphyllin A, deidaclin, epivolkenin and taraktophyllin, P. discophora tetraphyllin B and volkenin, and P. x violacea tetraphyllin B sulfate. The remaining species were noncyanogenic. The glycosides were identified by 1H and 13C NMR spectroscopy following isolation by reversed-phase preparative HPLC. From P. guatemalensis, a new glucoside named passiguatemalin was isolated and identified as a 1-(beta-D-glucopyranosyloxy)-2,3-dihydroxycyclopentane-1 carbonitrile. An isomeric glycoside was prepared by catalytic hydrogenation of gynocardin. alpha-Hydroxyamides corresponding to the cyanogenic glycosides were isolated from several Passiflora species. These alpha-hydroxyamides, presumably formed during processing of the plant material, behave as cyanogenic compounds when treated with commercial Helix pomatia crude enzyme preparation. Thus, the enzyme preparation appears to contain an amide dehydratase, which converts alpha hydroxyamides to cyanohydrins that liberate cyanide; this finding is of interest in connection with analysis of plant tissues and extracts using Helix pomatia enzymes. PMID- 11853746 TI - Phenolic compounds from Gastrodia rhizome and relaxant effects of related compounds on isolated smooth muscle preparation. AB - Gastrol (1), together with 10 known phenolic compounds, has been isolated from the MeOH extract of the rhizomes of Gastrodia elata Blume (Orchidaceae), and their structures were elucidated by detailed spectral analyses including by 2D NMR spectroscopic analyses. The relaxant effects of these constituents on smooth muscle preparations isolated from guinea-pig ileum were also studied in order to reveal their characteristic pharmacological activities. PMID- 11853747 TI - Antifungal amides from Piper arboreum and Piper tuberculatum. AB - In continuation of our study of the Piperaceae we have isolated several amides, mainly those bearing isobutyl, pyrrolidine, dihydropyridone and piperidine moieties. Bioactivity-guided fractionation of extracts from leaves of Piper arboreum yielded two new amides, N-[10-(13,14-methylenedioxyphenyl)-7(E),9(Z) pentadienoyl]-pyrrolidine (1), arboreumine (2) together with the known compounds N-[10-(13,14-methylenedioxyphenyl)-7(E)-pentaenoyl]-pyrrolidine (3) and N-[10 (13,14-methylenedioxyphenyl)-7(E),9(E)-pentadienoyl]-pyrrolidine (4). Catalytic hydrogenation of 3 yielded the amide N-[10-(13,14-methylenedioxyphenyl) pentanoyl]-pyrrolidine (5). We also have isolated six amides (6-11) and two antifungal cinnamoyl derivatives (12, 13) from seeds and leaves of Piper tuberculatum. Compounds 1-11 showed antifungal activity as determined by direct bioautography against Cladosporium sphaerospermum while compounds 3-4 and 6-13 also showed antifungal activity against C. cladosporioides. PMID- 11853748 TI - Germacranolides from Artemisia myriantha and their conformation. AB - The CH(2)Cl(2) extract of the aerial parts of Artemisia myriantha afforded three germacranolides derived from 13-acetoxy-3beta-hydroxy-germacra-1(10)E,4E,7(11) trien-12,6alpha-olide, whose structures were elucidated by 2D-NMR spectroscopic analyses. Some conclusions are drawn about the possible conformations of the ten membered germacranolide ring system from the exchange peaks seen in the NOESY spectra, and an estimate is made of the energy barrier to ring-flipping from variable-temperature NOESY experiments. The conclusions reached were supported by molecular modeling studies and an NMR spectroscopic investigation of the commercially available germacranolide, parthenolide. PMID- 11853749 TI - Non-glycosidic iridoids from Cymbaria mongolica. AB - Six non-glycosidic iridoids, i.e. (1R,4S,4aS,7S,7aS)-7-hydroxyl-4-hydroxy- methyl 7-methyl-1-methoxyl-1,4,4a,7a-tetrahydrocyclopenta[e]pyran-3-one (1), (1S,4R,4aS,7S,7aS)-7-hydroxyl-4-hydroxymethyl-7-methyl-1-methoxyl-1,4,4a,7a tetrahydrocyclopenta[e]pyran-3-one (2), (1R,4R,4aS,7S,7aS)-7-hydroxyl-4-hydroxy methyl-7-methyl-1-methoxyl-1,4,4a,7a-tetrahydrocyclopenta[e]pyran-3-one (3), (1R, 4R, 4aS, 7aS)-4,7-dihydroxymethyl-1-methoxyl-1,4,4a,7a-tetrahydrocyclopenta-6 ene[e]pyran-3-one (4), (1R, 4R, 4aS, 7aS)-4,7-dihydroxymethyl-1-hydroxyl-1,4,4a, 7a-tetrahydrocyclopenta-6-ene[e]pyran-3-one (5), (1R, 4S, 4aS, 7aS)-4,7-dihydroxy methyl-1-methoxyl-1,4,4a,7a-tetrahydrocyclopenta-6-ene[e]pyran-3-one (6), as well as five known non-glycosidic iridoids mussaenin A (7), gardendiol (8), isoboonein (9), 4-epi-alyxialactone (10) and rehmaglutin D (11) have been isolated from the Chinese medicinal plant Cymbaria mongolica. Their structures were elucidated by spectroscopic methods. These compounds exhibit significant antitumor and antibacterial activity. PMID- 11853750 TI - Polyoxygenated cyclohexene derivatives from Ellipeiopsis cherrevensis. AB - Aerial parts of Ellipeiopsis cherrevensis contained the polyoxygenated cyclohexenes zeylenol, ferrudiol and three analogs, ellipeiopsols A, B and C. The C-1 stereochemistry of ferrudiol has been revised. PMID- 11853751 TI - Iridoid and phenolic glycosides from Morinda coreia. AB - From the leaves and branches of Morinda coreia, six compounds [yopaaosides A-C, 10-O-acetylmonotropein, 6-O-acetylscandoside and 3,4,5-trimethoxyphenyl 1-O-beta apiofuranosyl (1"-->6')-beta-glucopyranoside] have been isolated together with five known compounds. Structural elucidations were based on analyses of physical and spectroscopic data. PMID- 11853752 TI - Phenolic glycosides from Markhamia stipulata. AB - From the leaves and branches of Markhamia stipulata, five verbascoside derivatives (markhamiosides A-E), and one hydroquinone derivative (markhamioside F) were isolated together with 13 known compounds. Their structural elucidation was based on analyses of chemical and spectroscopic data. PMID- 11853753 TI - Phenolic glycosides from Barnettia kerrii. AB - From the leaves and branches Barnettia kerrii, two verbascoside derivatives (6"-O acetylverbascoside and 4"'-O-acetylverbascoside), as well as two phenolic glycosides (khaephuosides A and B) were isolated together with 15 known compounds. The structural elucidations were based on analyses of spectroscopic data. PMID- 11853754 TI - Blazeispirols B, C, E and F, des-A-ergostane-type compounds, from the cultured mycelia of the fungus Agaricus blazei. AB - Four new des-A-ergostane derivatives including blazeispirols B, C, E and F were isolated from the cultured mycelia of fungus Agaricus blazei Murill and were established to be (20S, 22R, 23R, 24S)-14beta,22: 22,25-diepoxy-5-methoxy-des-A ergosta-5,7,9,11-tetraen-23-ol; (20S, 22S, 23R, 24S)-14beta,22: 22,25-diepoxy-5 methoxy-des-A-ergosta-5,7,9-trien-23-ol; (20S, 22S, 23R, 24S)-14beta, 22: 22, 25 diepoxy-5-methoxy-des-A-ergosta-5,7,9,11-tetraene-19,23-diol and (20S, 22S, 23R, 24S)-14beta,22: 22,25-diepoxy-des-A-ergosta-5,7,9-triene-5,23-diol by comparison of extensive 1D and 2D NMR spectral data with that of blazeispirol A. PMID- 11853755 TI - Animal models in research on retinal degenerations: past progress and future hope. AB - The retinal degenerations (RDs) are a family of inherited retinal degenerative diseases (dystrophies) that lead to vision loss. Although phenotypically very different, the RDs have several characteristics in common. They all are caused by gene mutations or at least have a genetic component in the etiology. They all lead to photoreceptor dysfunction, many leading to the death of both rod and cone photoreceptors. The mechanism of cell death in most of the RDs seems to be through the process of apoptosis. It is estimated that more than fifteen million people around the world have vision loss due to an inherited RD. Many of these are patients with the dry form of age-related macular degeneration (AMD) who retain partial functional vision. However, some have other degenerative conditions such as retinitis pigmentosa, Leber congenital amaurosis or wet AMD and can suffer from severe vision loss or total blindness. PMID- 11853756 TI - Functionally rodless mice: transgenic models for the investigation of cone function in retinal disease and therapy. AB - Two genetically engineered strains of mice were used to characterize murine cone function electroretinographically, without interference of rod-driven responses: (1) mice with a deletion of the gene for the rod transducin alpha-subunit (transducin alpha-/-), and (2) mice with rod arrestin deleted (arrestin -/-). In the first three months of age, both strains have a normal complement of rods and normal rod structure, but transducin alpha-/- mice have no rod-driven responses to light, while rod-driven activity of arrestin -/- mice can be suppressed by a single intense flash for hours. In response to intense flashes the electroretinograms of these strains of mice showed a readily identifiable, pure cone a-wave of approximately 10 microV saturating amplitude. A 530 nm background that saturates rod responses of wild type mice was found to desensitize the b wave responses of mice of both transgenic lines, whether the b-waves were driven by photons captured by M- or UV-cone pigments. The desensitizing effect of the 530 nm background on UV-pigment driven responses provides new evidence in support of the hypothesis of functional co-expression of the M-pigment in cones expressing primarily the UV-pigment. PMID- 11853757 TI - The carboxyl-terminal domain is essential for rhodopsin transport in rod photoreceptors. AB - The role of the carboxyl-terminal domain in rhodopsin transport was investigated using transgenic mice expressing a rhodopsin truncation mutant lacking the terminal 15 amino acids (S334ter). It was previously shown that S334ter translocates to the outer segment in the presence of endogenous rhodopsin. We now show that in the absence of endogenous rhodopsin S334ter mis-localizes to the plasma membrane and fails to reconstitute outer segment structures. Surprisingly, this mis-localization does not affect photoreceptor cell survival. These results provide further evidence on the important role of the COOH-terminal domain in rhodopsin trafficking and demonstrate an absolute requirement of this domain for correct vectorial transport of rhodopsin in rod photoreceptors. PMID- 11853758 TI - Retinal ablation and altered lens differentiation induced by ocular overexpression of BMP7. AB - The alphaA-crystallin promoter was used to target expression of bone morphogenetic protein 7 (BMP7) to lens fiber cells in transgenic mice. Surprisingly, lens-specific expression of BMP7 induced widespread apoptosis and rapid ablation of the neural retina in multiple families. Subsequent to retinal ablation, the lens bow region shifted posteriorly until lens epithelial cells completely enveloped the lens. Lens-specific expression of FGF3 was found to rescue the loss of fiber cell differentiation. Our results show that elevated BMP7 levels can induce rapid retinal degeneration accompanied by disruption of the endogenous ocular system for fiber cell induction. PMID- 11853759 TI - The positive role of the carboxyl terminus of the gamma subunit of retinal cGMP phosphodiesterase in maintaining phosphodiesterase activity in vivo. AB - The inhibitory rod cyclic GMP-phosphodiesterase gamma subunit, PDEgamma, is a key component of the photoresponse and is required to support rod integrity. Pdeg(tm1)/Pdeg(tm1) mice that lack PDEgamma due to a targeted disruption of the gene encoding PDEgamma, (Pdeg) suffer from a very rapid and severe photoreceptor degeneration. Previously, deletions in the carboxyl-terminal domain of PDEgamma blocked its ability to inhibit trypsin-activated PDE activity, in vitro. In other words, these mutations eliminated PDEgamma's control on the catalytic activity of PDEalpha and PDEbeta. To study the in vivo effects resulting from the deletion of the last seven amino acids of the PDEgamma carboxyl terminal, this PDEgamma allele (Del7C) was introduced as a transgene Pdeg(tm1)/Pdeg(tm1) mice. These animals could only synthesize transgenic mutant PDEgamma. The mutant retinas were expected to display a higher basal level of PDE activity and lower cGMP levels in light and darkness than the PDEgamma knockout mice, which would allow the rescue of their photoreceptors. Instead, our results showed that the Del7C transgene could not complement the Pdeg(tm1)/Pdeg(tm1) mutant for photoreceptor survival. In fact, animals carrying the Del7C transgene have low PDE activity as well as reduced PDEalpha and PDEbeta content. PMID- 11853760 TI - Inactivation of the Basigin gene impairs normal retinal development and maturation. AB - 5A11/Basigin is an immunoglobulin-like glycoprotein expressed on the surface of Muller cells, the apical and basal surfaces of the retinal pigmented epithelium, and photoreceptor cell bodies and their inner segments. Disruption of the 5A11/Basigin gene in the mouse results in photoreceptor degeneration and a corresponding decrease in electroretinogram amplitudes in mature mice. The purpose of this study was to examine the electrophysiology of the 5A11/Basigin null mouse retina at earlier ages than previously examined. Although the architecture of the 5A11/Basigin null mouse retina appears normal, the ERG amplitudes are severely depressed at eye opening, indicating failure in retinal maturation. PMID- 11853761 TI - Transport to the photoreceptor outer segment by myosin VIIa and kinesin II. AB - This paper reviews our studies in which we have used mutant mice to investigate the roles of myosin VIIa and kinesin II in the transport of proteins to the photoreceptor outer segment. These studies suggest that both motors participate in moving opsin along the connecting cilium. Given the velocities measured for these motors in vitro, it is predicted that the resulting concentration of opsin in the plasma membrane of the connecting cilium is surprisingly low. PMID- 11853762 TI - Mouse eye gene microarrays for investigating ocular development and disease. AB - Microarray technology can facilitate simultaneous expression analysis of thousands of genes and assist in delineating cellular pathways involved in development or disease pathogenesis. Since public databases and commercial cDNA microarrays have an under-representation of eye-expressed genes, we generated over 3000 expressed sequence tags from three unamplified mouse eye/retina cDNA libraries. These eye-expressed genes were used to produce cDNA microarrays. Methodology for printing of slides, hybridization, scanning and data analysis has been optimized. The I-gene microarrays will be useful for establishing expression profiles of the mouse eye/retina and provide a resource for defining molecular pathways involved in development, aging and disease. PMID- 11853763 TI - Prospects for relevant glaucoma models with retinal ganglion cell damage in the rodent eye. AB - Retinal ganglion cell (RGC) death is the end result of practically all diseases of the optic nerve, including glaucomatous optic neuropathy. Understanding the factors determining susceptibility of the retina or the optic nerve to glaucomatous damage, and the means to prevent it, requires good animal models. Here we review the different, current models in rodents that have been used to study RGC damage, discuss their value, and their adequacy as models for human glaucoma. PMID- 11853764 TI - Generating mouse models of retinal disease using ENU mutagenesis. AB - We used the chemical mutagen, N-ethyl-N-nitrosourea, to induce random point mutations in the germline of the mouse strain C57BL/6 in order to generate models of retinal diseases. 1163 mutagenised first generation mice produced using this approach were examined for eye abnormalities. Approximately one-third (412) presented with some form of ocular abnormality. Most changes were unilateral and confined to the anterior segment of the eye. Less than 10% (44) of identified changes affected the posterior segment of the eye. 21 mice with varying ocular abnormalities, including 17 with retinal changes, were bred to produce second generation mice to confirm genetic inheritance. Genetic inheritance was confirmed in several of these lines including three with retinal changes. PMID- 11853765 TI - Immunobiology and privilege of neuronal retina and pigment epithelium transplants. AB - Despite the existence of ocular immune privilege, immune rejection may be a barrier to successful retinal transplantation. We have examined in mice the extent to which the subretinal space (SRS) is an immune privileged site, and whether retinal pigment epithelium and neuronal retinal tissue have properties of immune privileged tissues. We report that (1) The SRS is an immune privileged site; (2) Neonatal RPE is an immune privileged tissue; (3) Neuronal retina is a partially immune privileged tissue; and (4) Microglia within neonatal neural retina grafts promote photoreceptor differentiation, become activated, and induce sensitization of the recipient and serve as targets of immune rejection. PMID- 11853766 TI - Loss of the phospholipase C gene product induces massive endocytosis of rhodopsin and arrestin in Drosophila photoreceptors. AB - Previously we have shown that a subset of visual transduction mutants in Drosophila melanogaster induce the formation of stable complexes between rhodopsin and arrestin. One such mutant is in a visual system-specific phospholipase C (PLC). The rhodopsin/arrestin complexes generated in PLC mutants induce massive retinal degeneration. Here we demonstrate that both arrestin and rhodopsin undergo light-dependent endocytosis in a PLC mutant background. Interestingly, the internalized rhodopsin is rapidly degraded, but the arrestin is fully stable. The data are discussed with respect to mechanisms of arrestin mediated endocytosis and human retinal disease. PMID- 11853767 TI - Rpr- and hid-driven cell death in Drosophila photoreceptors. AB - The reaper (rpr) and head involution defective (hid) genes mediate programmed cell death (PCD) during Drosophila development. We show that expression of either rpr or hid under control of a rhodopsin promoter induces rapid cell death of adult photoreceptor cells. Ultrastructural analysis revealed that the dying photoreceptor cells share morphological features with other cells undergoing PCD. The anti-apoptotic baculoviral P35 protein acts downstream of hid activity to suppress the photoreceptor cell death driven by rpr and hid. These results establish that the Drosophila photoreceptors are sensitive to the rpr- and hid driven cell death pathways. PMID- 11853768 TI - Retinal degeneration mutants in the mouse. AB - The Jackson Laboratory, having the world's largest collection of mouse mutant stocks and genetically diverse inbred strains, is an ideal place to look for genetically determined eye variations and disorders. Through ophthalmoscopy, electroretinography and histology, we have discovered disorders affecting all aspects of the eye including the lid, cornea, iris, lens and retina, resulting in corneal disorders, cataracts, glaucoma and retinal degenerations. Mouse models of retinal degeneration have been investigated for many years in the hope of understanding the causes of photoreceptor cell death. Sixteen naturally occurring mouse mutants that manifest degeneration of photoreceptors in the retina with preservation of all other retinal cell types have been found: retinal degeneration (formerly rd, identical with rodless retina, r, now Pde6b(rd1)); Purkinje cell degeneration (pcd); nervous (nr); retinal degeneration slow (rds, now Prph(Rd2)); retinal degeneration 3 (rd3); motor neuron degeneration (mnd); retinal degeneration 4 (Rd4); retinal degeneration 5 (rd5, now tub); vitiligo (vit, now Mitf(mi-vit)); retinal degeneration 6 (rd6); retinal degeneration 7 (rd7, now Nr2e3(rd7)); neuronal ceroid lipofuscinosis (nclf); retinal degeneration 8 (rd8); retinal degeneration 9 (Rd9); retinal degeneration 10 (rd10, now Pde6b(rd10)); and cone photoreceptor function loss (cpfl1). In this report, we first review the genotypes and phenotypes of these mutants and second, list the mouse strains that carry each mutation. We will also provide detailed information about the cpfl1 mutation. The phenotypic characteristics of cpfl1 mice are similar to those observed in patients with complete achromatopsia (ACHM2, OMIM 216900) and the cpfl1 mutation is the first naturally-arising mutation in mice to cause cone-specific photoreceptor function loss. cpfl1 mice may provide a model for congenital achromatopsia in humans. PMID- 11853769 TI - Forward and reverse genetic approaches to the analysis of eye development in zebrafish. AB - The zebrafish has been established as a mainstream research system, largely due to the immense success of genetic screens. Over 2000 mutant alleles affecting zebrafish's early development have been isolated in two large-scale morphological screens and several smaller efforts. So far, over 50 mutant strains display retinal defects and many more have been shown to affect the retinotectal projection. More recently, mutant isolation and characterization have been successfully followed by candidate and positional cloning of underlying genes. To supplement forward genetic mutational analysis, several reverse genetic techniques have also been developed. These recent advances, combined with the genome project, have established the zebrafish as one of the leading models for studies of visual system development. PMID- 11853770 TI - Zebrafish mutagenesis yields eye morphological mutants with retinal and lens defects. AB - A chemical mutagenesis to identify zebrafish eye morphological mutants was performed by screening F(3) larvae at 5 and 7 days post-fertilization (dpf) for changes in eye or pupil size. Based on histological analysis, four different phenotypic classes were obtained. The two Class I and three Class II mutants are all characterized by small eyes and exhibit defects in early retinal development or unregulated cell death, respectively. The single Class III mutant has reduced ocular pigmentation. The three Class IV mutants display defects in the ocular lens, including one mutant line with normal sized eyes and pupils that develops lens opacity at 7 dpf. PMID- 11853771 TI - Kinetics of transgene expression in mouse retina following sub-retinal injection of recombinant adeno-associated virus. AB - Using confocal microscopy we have examined in detail the temporal and spatial pattern of green fluorescent protein expression following sub-retinal injection of recombinant adeno-associated virus (rAAV) in the mouse and have determined the effect of viral titre on the number and type of cells transduced. Our results suggest that some transgene expression occurs as early as three days after injection, and that transgene expression occurs beyond the area of retinal detachment. Vector titre appears to have a substantial effect on both transduction efficiency and the speed of onset of photoreceptor cell transduction. Our data suggests that we have not yet reached the limits of photoreceptor transduction efficiency using AAV vectors. An increase in titre could still lead to an improved transduction efficiency and faster onset of photoreceptor transduction. We failed to detect transfected cones even in areas where nearly 100% of the rods were transduced, but we found efficient and sustained RPE transduction. PMID- 11853773 TI - Color categories revealed by non-negative matrix factorization of Munsell color spectra. AB - Non-negative matrix factorization (NMF, Nature 401 (1999) 788-791) is a method to derive non-negative basis functions for sets of data that are inherently non negative, such as color spectra. We applied NMF to Munsell color spectra and investigated the color names associated with the non-negative basis functions. NMF yields basis functions compatible with established color naming categories. PMID- 11853772 TI - Lentiviral transduction of green fluorescent protein in retinal epithelium: evidence of rejection. AB - This paper demonstrates lentiviral transduction of the humanized form of the Aequoria victoria gene for green fluorescent protein (GFP) into human fetal retinal pigment epithelium (RPE) in vitro and rabbit RPE in vivo. In vitro GFP expression of cultured human fetal RPE begins within two to three days after 12 16 h of maintained exposure to the virus at titers of 10(8)-10(9) infectious units (IU)/ml. Both stationary and dividing cells are transduced using a lenti viral vector with a cytomegalovirus (CMV) promoter. Expression remains stable for at least three to four months without evidence of toxicity and continues through cell division. In vivo expression is followed non-invasively in rabbit eye using a scanning laser ophthalmoscope (SLO), which can detect single fluorescing retinal cells. In vivo expression begins within a few days after a viral solution is introduced into the subretinal space. A solution of 10(9) IU/ml produces fluorescence within three to four days. Less concentrated solutions lead to slower and less expression. No expression is detectable at concentrations of 10(6) IU/ml. Within one to two weeks after introduction of the viral solution, there is evidence of rejection seen by SLO as a loss of GFP fluorescence and disruption of the RPE. Histology shows damage to the RPE layer and monocytic cell infiltrates in the choroid and subretinal space within the area receiving the viral solution. Strong GFP expression leads to rejection within two weeks. With less expression, rejection is delayed and in some cases undetectable for at least six months. If the GFP gene is not included in the viral vector or if the viral concentration is insufficient to produce detectable GFP expression, rejection is not seen. Using a rhodopsin promoter or injecting the virus intra rather than subretinally produces weak expression and no rejection. Lentivirus can induce expression of a foreign gene in the RPE. Viral induced transduction and GFP expression have no effect on the viability of the RPE in vitro. Continued expression of GFP after cell division implies chromosomal integration of the gene. In vivo expression of GFP in RPE encounters rejection. Rejection may not occur with low GFP expression. The latter occurs with low viral titers, a rhodopsin promoter or intra-retinal injection of viral solution. The results are relevant to gene therapy in retina when gene transduction leads to the expression of foreign proteins. PMID- 11853774 TI - Comparison of color and luminance vision on a global shape discrimination task. AB - We compared the performances of the blue-yellow, red-green and luminance systems on a shape discrimination task. Stimuli were radial frequency patterns (radially modulated fourth derivative of a Gaussian) with a peak spatial frequency of 0.75 cpd. Stimuli isolated the chromatic (red-green and blue-yellow) and achromatic post-receptoral mechanisms. We showed that in all cases performance, measured as a radial modulation threshold for discrimination between a circular and non circular stimulus, improves with contrast. Performance was compared across radial frequencies with contrast matched in multiples of stimulus detection threshold. We find that blue-yellow color system performs the worse on this shape discrimination task, followed by the red-green, with the achromatic system performing best. The average difference is a factor of 2 between achromatic and blue-yellow performance, and a factor of 1.7 between red-green and achromatic. Despite these performance losses, chromatic shape discrimination can still reach hyperacuity performance levels. In a secondary experiment we contrast modulate the radial contour to eliminate either the "corners" or "sides" of an RF4 (square) pattern. We find that for the achromatic system, the sides are more important for the task than the corners. However, for the chromatic system, removal of sides or corners produces similar performance deficits. We conclude that color vision has a selective although relatively mild deficit for two dimensional form perception. PMID- 11853775 TI - Bivectorial transparent stimuli simultaneously adapt mechanisms at different levels of the motion pathway. AB - The motion aftereffect (MAE) to drifting bivectorial stimuli, such as plaids, is usually univectorial and in a direction opposite to the pattern direction of the plaid. This is true for plaids that are perceived as coherent, but also for other plaids which are seen as transparent for most or all of the adaptation period. The underlying mechanisms of this MAE are still not well understood. In order to assess these mechanisms further, we measured static and dynamic MAEs and their interocular transfer (IOT). Adaptation stimuli were plaids with small (coherent) and large (transparent) angles between the directions of the component gratings and a horizontal grating, which were adjusted in spatial frequency and drift velocity so that the pattern speed and vertical periodicity remained constant. Test stimuli were horizontal static or counterphasing gratings with the same periodicity as the adaptation stimuli. MAE duration was measured for monocular, binocular and IOT conditions. All static MAEs were smallest for the transparent plaid and largest for the grating, while all dynamic MAEs were constant across adaptation stimuli. IOT was twice as big for dynamic MAEs as for static MAEs, and did not vary with the adaptation stimuli. Other adaptation stimuli were plaids that differed in intersection luminance, contrast or spatial frequency, resulting in different amounts of perceived coherence. MAEs and IOT did not vary with perceived coherence. The results suggest that the MAE for bivectorial stimuli consists of low-level adaptation (dependent on local component properties, small IOT), as well as high-level adaptation (dependent on global integrated pattern properties, large IOT), which can be measured independently with static and dynamic test stimuli. PMID- 11853776 TI - Symmetry perception: a novel approach for biological shapes. AB - The majority of quantitative studies on symmetry perception have employed random dot patterns, yet symmetrical random patterns are not common in nature. Here we explore symmetry perception utilizing sums of radial frequency (RF) patterns to define complex shapes. When a pair of RF patterns with different frequencies are added, the relative phase of the two components provides a precise measure of the degree of deviation from bilateral symmetry. Sums of RF2-RF7 define such diverse biological shapes as human heads, animal heads, torsos, and many fruit, so discrimination of symmetries defined by these patterns is highly relevant to biological vision. Here we show that symmetry discrimination during brief presentations is best for RF2+RF3 but becomes impossible for RF2+RF7. Further experiments demonstrate that the underlying neural mechanisms differ from those involved in random dot symmetry detection. These results were used to predict symmetry thresholds for deviations from bilateral symmetry of head shapes based on a principal components analysis of 30 female heads. Human V4 is hypothesized to be the site for symmetry discrimination of RF patterns but not of random dot patterns. PMID- 11853777 TI - Spatial frequency discrimination: a comparison of achromatic and chromatic conditions. AB - In this study, we have compared foveal SF discriminations for luminance and color defined stimuli using two different tasks (criteria): in criterion-A, the discrimination is based on spatial (size of the stimuli) and/or spatial frequency; in criterion-B, it is based on apparent motion (contraction/expansion). We used high contrast (75%) spatially localized D6 stimuli and cosine gratings (0.25-9.5 cpd). The SF discrimination was measured by the method of constant stimuli with a two-interval forced-choice procedure. Data show that: (i) for criterion-A, the discrimination thresholds for color stimuli were lower than that for luminance stimuli at low SFs, but similar or higher at higher SFs; for criterion-B, the thresholds to chromatic stimuli were higher than that to achromatic stimuli for all SFs; (ii) SF discrimination was best at inter stimulus-interval (ISI) of about 200 ms for color stimuli and at ISI of 0 ms for luminance stimuli; (iii) SF discrimination got better with stimulus duration and reached to plateau at 200 ms (or more) for color stimuli and at 67 ms (or more) for luminance stimuli; (iv) SF discrimination threshold (mean Delta(f)=0.19 octaves) is about one-tenth of the full bandwidth (mean=1.96 octaves) of SF tuned mechanisms and is in hyperacuity range; both (discrimination and hyperacuity) can be explained by the relative activities within a population of tuned mechanisms. We conclude that color and luminance SF discrimination thresholds have a different SF dependence. While color appears to perform better than luminance vision at low SFs, this effect is lost or even reversed at high SFs. Data imply that color and form interact, but color and motion are largely segregated (i.e. they weakly interact). PMID- 11853778 TI - Orientation sensitivity in human visual motion processing. AB - Orientation tuning of receptive fields is well documented in the spatial domain, but considerable variability exists amongst published estimates of orientation sensitivity of motion receptive fields. We used a two-frame motion sequence, in which one frame was binary noise and the other was a horizontally displaced and filtered version of the same noise field, to examine the orientation sensitivity of human motion mechanisms. Initially, orientations orthogonal to the direction of motion were removed from each filtered frame. Observers indicated perceived direction of motion in a single interval, binary choice task. D(max) was determined for different amounts of removed orientations, and found to remain constant across the removal of energy up to approximately +/-60 deg from vertical. In a second experiment, the orientations removed were now parallel to the direction of motion of the stimulus. D(max) fell as a cosine function with increasing removal of orientation information, in agreement with off-orientation looking or matched filtering predictions. The two experiments show the presence of mechanisms both broadly tuned and more narrowly tuned for orientation. A control experiment introduced an interstimulus interval between the two frames of our motion sequence. Performance on the direction discrimination task was severely degraded, indicating that the original results are not explicable in terms of a feature-tracking or long-range motion process. The presence of both broadly and narrowly tuned mechanisms implies multiple possible solutions to the processing of coherent plaid motion. PMID- 11853779 TI - A dynamical model of saccade generation in reading based on spatially distributed lexical processing. AB - The understanding of the control of eye movements has greatly benefited from the analysis of mathematical models. Currently most comprehensive models include sequential shifts of visual attention. Here we propose an alternative model of eye movement control, which includes three new principles: spatially distributed lexical processing, a separation of saccade timing from saccade target selection, and autonomous (random) generation of saccades with foveal inhibition. These three features provide a common control mechanism for fixations, refixations, and regressions. Consequently, the model is called SWIFT (Saccade-generation with inhibition by foveal targets). Results from numerical simulations are in good agreement with effects of word frequency on single-fixation, first-fixation, and gaze durations as well as fixation and word skipping probabilities in first-pass analysis. The model inherently produces complex eye movement patterns including refixations and regressions due to its underlying dynamical principles. PMID- 11853780 TI - Saccadic latency effects of progressively deleting stimulus offsets and onsets. AB - We designed two extensions of Saslow's well-known gap and overlap conditions that require increased voluntary effort because of the progressive elimination of target onsets and fixation point offsets, and obtained repeatable data obeying simple numerical relations. For each of the five stimulus lighting conditions, saccadic latency was measured as a function of the retinal eccentricity or displacement of the target. Latencies were fitted by a serial processing model in which the visually guided minimum tracking latency VGL(min) is supplemented by two types of delay, dubbed 'unlock' and 'target', that can be either short or long ('direct' or 'indirect'), depending on the conditions. There are two findings: (1) The model has utility. The rank order of saccadic latencies for the five stimulus lighting conditions was constant across all subjects, sessions and eccentricities in the range 7.5'-6 degrees left or right. For pooled data, and the saccadic latency plateau (1-6 degrees), the model was also within +/-3 ms of the mean latencies. (2) Latencies of tiny saccades to intrafoveolar stimulation (7.5-45') were invariably long in all five stimulus conditions. One factor here must be the experimentally measured local prolongation of VGL(min). PMID- 11853781 TI - Rules for combining the outputs of local motion detectors to define simple contours. AB - We know something about the fidelity with which motion can be detected in local regions of the visual field but nothing about how these local motion signals are combined across space to define contours. To investigate such linking rules, we measured the detectability of motion-defined contours using an adaptation of the paradigm of Field, Hayes, and Hess (Vision Research, 33 (1993) 173) in which subjects are asked to detect the presence of simple contours defined solely by local motion direction that are embedded in a field of otherwise random local motions. We show that contours defined by motion whose direction is along the contour are more detectable than contours defined by motions of any common direction. Furthermore, the contour configuration is important in that straight and moderately curved contours, though not highly curved ones, can support this specialized form of motion integration. PMID- 11853782 TI - Orientation characteristics of a mechanism in the human visual system sensitive to cyclopean form. AB - Human observers viewed two superimposed cyclopean gratings in dynamic random noise. Neither grating was visible monocularly. The disparities of the two gratings were counterphase-modulated at different temporal frequencies. We recorded frequency domain brain responses at ultra-high frequency resolution (0.008 Hz) by means of nondestructive zoom-FFT. The response to the superimposed gratings was quite different to the sum of responses to the two individual gratings. We used this strong nonlinearity to investigate the orientation tuning of the cyclopean mechanism that responded to the gratings. One grating had a fixed orientation while the other was rotated. Half-sensitivity full bandwidth was greater than 90 degrees. This is considerably greater than the corresponding bandwidth (30 degrees) for luminance-defined (LD) gratings [Vis. Res. 27 (1987) 2181]. Furthermore, the cyclopean nonlinearity was different from the corresponding nonlinearity for LD gratings. Brain responses to a single cyclopean grating showed an inverse oblique effect over a 1.2-1.9 degrees range of bar widths, but not for smaller or larger bar widths. PMID- 11853783 TI - The effect of visual field defects on eye movements and practical fitness to drive. AB - Eye movements of subjects with visual field defects due to ocular pathology were monitored while performing a dot counting task and a visual search task. Subjects with peripheral field defects required more fixations, longer search times, made more errors, and had shorter fixation durations than control subjects. Subjects with central field defects performed less well than control subjects although no specific impairment could be pinpointed. In both groups a monotonous relationship was observed between the visual field impairment and eye movement parameters. The use of eye movement parameters to predict viewing behavior in a complex task (e.g. driving) was limited. PMID- 11853784 TI - Focusing on health in the crisis in Zimbabwe. PMID- 11853785 TI - Role of D-dimers in diagnosis of venous thromboembolism. PMID- 11853786 TI - Resistance to targeted therapy in leukaemia. PMID- 11853787 TI - The doctor's role in advocacy. PMID- 11853788 TI - Follow-up of children born after in-vitro fertilisation. PMID- 11853789 TI - Sheila Sherlock's legacy. PMID- 11853790 TI - Neurological sequelae in children born after in-vitro fertilisation: a population based study. AB - BACKGROUND: There is an absence of population-based long-term studies on the risk of neurological sequelae in children born after in-vitro fertilisation (IVF). Our aim was to compare the frequency of such problems between IVF-born children and controls. METHODS: We did a population-based retrospective cohort study in which we compared development of neurological problems in 5680 children born after IVF, with 11360 matched controls. For 2060 twins born after IVF, a second set of controls (n=4120), all twins, were selected. We obtained data on neurological problems from the records of the Swedish habilitation centres. FINDINGS: Children born after IVF are more likely to need habilitation services than controls (odds ratio 1.7, 95% CI 1.3-2.2). For singletons, the risk was 1.4 (1.0-2.1). The most common neurological diagnosis was cerebral palsy, for which children born after IVF had an increased risk of 3.7(2.0-6.6), and IVF singletons of 2.8 (1.3-5.8). Suspected developmental delay was increased four-fold (1.9-8.3) in children born after IVF. Twins born after IVF did not differ from control twins with respect to risk of neurological sequelae. Low-birthweight and premature infants were more likely to need habilitation than fullterm babies. Maternal age did not affect risk. INTERPRETATION: Our study suggests that children born after IVF have an increased risk of developing neurological problems, especially cerebral palsy. These risks are largely due to the high frequency of twin pregnancies, low birthweight, and prematurity among babies born after IVF. To limit these risks, we recommend that only one embryo should be transferred during IVF. PMID- 11853791 TI - Detection and treatment of claudication due to functional iliac obstruction in top endurance athletes: a prospective study. AB - BACKGROUND: Endurance athletes often have restrictions in flow in their iliac arteries during exercise. Such restrictions have previously been ascribed solely to intravascular lesions. We postulate that flow could also be restricted by functional kinking in the arteries, and that surgical release of these kinks might be an effective treatment. METHODS: We prospectively studied 80 endurance athletes who had complaints suggestive of flow restriction in the iliac arteries of one (n=74) or both (6) legs (total 92 legs). Using vascular diagnostic tools, we examined athletes while they were doing activities that often provoke flow restrictions. Restrictions were determined by measurement of systolic pressure in the ankle after exercise; peak systolic velocities were measured with echo doppler. Kinks were detected with echo-doppler and magnetic-resonance angiography. When functional kinking was diagnosed as the cause of the restriction, the athlete was offered surgery to release the iliac arteries, as part of our prospective study. FINDINGS: We recorded flow restrictions in the iliac arteries of 58 of 92 (63%) legs. In 40 of these legs (69%), kinks were the most important cause of the restriction, making these legs suitable for surgical release. We operated on 23 of 58 (40%) legs. All athletes who had an operation subjectively improved. Maximum workload in a cycling test and ankle pressure significantly improved after the operation. 20 (87%) athletes were able to successfully return to their desired high level of competition. INTERPRETATION: Our sports-specific protocol is effective in detecting kinking of the iliac arteries as a cause for flow restriction in athletes who have few intravascular abnormalities when investigated with conventional vascular diagnostic tools. Surgical treatment directed at the kinking was less invasive and therefore a better alternative to vascular reconstruction in these athletes. PMID- 11853792 TI - Clinical picture: half a brain. PMID- 11853793 TI - Chloramphenicol versus benzylpenicillin and gentamicin for the treatment of severe pneumonia in children in Papua New Guinea: a randomised trial. AB - BACKGROUND: Pneumonia is the most frequent cause of child mortality in less developed countries. We aimed to establish whether the combination of benzylpenicillin and gentamicin or chloramphenicol would be better as first-line treatment in children with severe pneumonia in Papua New Guinea. METHODS: We did an open randomised trial in which we enrolled children aged 1 month to 5 years of age who fulfilled the WHO criteria for very severe pneumonia and who presented to hospitals in two provinces. Children were randomly assigned to receive chloramphenicol (25 mg/kg 6 hourly) or benzylpenicillin (50 mg/kg 6 hourly) plus gentamicin (7.5 mg/kg daily) by intramuscular injection. The primary outcome measure was a good or an adverse outcome. FINDINGS: 1116 children were enrolled; 559 children were treated with chloramphenicol and 557 with benzylpenicillin and gentamicin. At presentation the median haemoglobin oxygen saturation was 71% (IQR 57-77) for those allocated chloramphenicol and 69% (55-77) for those allocated penicillin and gentamicin. 147 (26%) children treated with chloramphenicol and 123 (22%) treated with penicillin and gentamicin had adverse outcomes (p=0.11). 36 children treated with chloramphenicol and 29 treated with penicillin and gentamicin died. More children treated with chloramphenicol than penicillin and gentamicin represented with severe pneumonia within 1 month of hospital discharge (p=0.03). INTERPRETATION: For children with severe pneumonia in less-developed countries the probability of a good outcome is similar if treated with chloramphenicol or with the combination of benzylpenicillin and gentamicin. PMID- 11853794 TI - Relation between resistance of Philadelphia-chromosome-positive acute lymphoblastic leukaemia to the tyrosine kinase inhibitor STI571 and gene expression profiles: a gene-expression study. AB - BACKGROUND: The ABL tyrosine kinase inhibitor STI571 is a promising agent for treatment of advanced Philadelphia-chromosome-positive (Ph+) acute lymphoblastic leukaemia. However, resistance to this drug develops within a few months in most patients. We aimed to predict resistance to STI571. METHODS: Total RNA was extracted from 25 bone-marrow samples from 19 patients with Ph+ acute lymphoblastic leukaemia who were enrolled into a phase II study. 17 samples were obtained before STI571 treatment was started: ten from individuals who were classified as good responders to STI571 (sensitive), and seven from individuals who did not to respond to STI571 (primary resistance). Eight samples were obtained from patients during treatment with STI571. We analysed six matched samples, which were obtained before and during treatment with STI571. Oligonucleotide microarray analysis of samples was done with high-density microarrays. FINDINGS: We identified 95 genes whose expression could be used to predict sensitivity of leukaemic cells to STI571. On this basis, all the STI571 sensitive samples could clearly be distinguished from the resistant cases. 56 highly differentially expressed genes were identified in leukaemic cells that were secondarily resistant to STI571. Resistant leukaemic cells expressed high levels of Bruton's tyrosine kinase and two ATP synthetases (ATP5A1 and ATP5C1), and showed significantly reduced expression of the proapoptotic gene BAK1 and the cell-cycle control gene p15 INK4b. INTERPRETATION: We have shown the clinical relevance of gene expression data for the pretreatment assessment of acute lymphoblastic leukaemia. Our results have implications for future clinical studies of tyrosine kinase inhibitors. PMID- 11853795 TI - BCR-ABL gene mutations in relation to clinical resistance of Philadelphia chromosome-positive leukaemia to STI571: a prospective study. AB - BACKGROUND: BCR-ABL, a constitutively activated tyrosine kinase, is the oncogene that causes Philadelphia-chromosome-positive (Ph+) leukaemia. STI571, a competitive inhibitor at the ATP-binding site of BCR-ABL, has been shown to have high activity in this type of leukaemia. However, most patients with advanced disease relapse despite continued treatment with STI571. We aimed to find out whether point mutations in BCR-ABL cause resistance to STI571. METHODS: We analysed clinical samples from eight patients resistant to STI571-who had advanced-stage Ph+ leukaemia-for mutations within the ATP-binding site and activation loop of BCR-ABL. Analysis was done before treatment with STI571 and at time of relapse. FINDINGS: We identified five distinct point mutations in the BCR ABL kinase domain in seven patients. All point mutations arose at positions that have proved to be important for drug binding and have conferred resistance to STI571 in vitro. All patients with mutations had lymphoid leukaemia. INTERPRETATION: Different mutations within the kinase domain of BCR-ABL can be responsible for refractoriness of Ph+ leukaemia to STI571. Mutation in the BCR ABL kinase domain might be a frequent mechanism of STI571 resistance in lymphoid disease. PMID- 11853796 TI - A man with a massive uvula. PMID- 11853797 TI - Reflux of gastric juice and glue ear in children. AB - Otitis media with effusion (glue ear) is the most frequent cause of deafness in children. We investigated the role of gastric juice reflux in this disease. We measured pepsin concentrations in middle ear effusions from children using ELISA and enzyme activity assays. 45 (83%) of 54 effusions contained pepsin/pepsinogen at concentrations of up to 1000-fold greater than those in serum. Our data suggest that reflux of gastric juice could be a major cause of glue ear in children. PMID- 11853798 TI - Blood transfusion for severe anaemia in children in a Kenyan hospital. AB - Severe anaemia often secondary to malaria is a major contributor to child mortality in sub-Saharan Africa. We have confirmed that use of simple clinical and laboratory criteria can identify those children likely to benefit most from treatment. We have also shown that the speed of response may be critical. The quality and capacity of blood transfusion services could therefore have a major, direct effect on inpatient child mortality. PMID- 11853799 TI - Desmopressin for nocturnal enuresis in nephrogenic diabetes insipidus. AB - We have investigated two unrelated families, in which two children had inherited primary nocturnal enuresis, and nephrogenic diabetes insipidus caused by new mutations in the aquaporin-2 gene (AQP2). The mutant AQP2 proteins were inactive, suggesting that administration of desmopressin could not concentrate the urine in these patients. However, treatment with desmopressin resolved primary nocturnal enuresis completely. This observation questions the notion that desmopressin resolves primary nocturnal enuresis through pharmacological manipulation of renal concentrating ability only. Desmopressin might also act on extrarenal targets such as the central nervous system. PMID- 11853800 TI - Merkel cell carcinoma and HIV infection. AB - Merkel cell carcinoma (MCC) is a rare skin cancer that occurs more frequently after organ transplantation or B-cell malignancy, conditions of suppressed or disordered immunity. To assess further whether immune suppression increases MCC risk, we studied its occurrence in a cohort of 309365 individuals with acquired immunodeficiency syndrome (AIDS) by using linked AIDS and cancer registries. We identified six cases of MCC, corresponding to a relative risk of 13.4 (95% CI 4.9 29.1) compared with the general population. These results suggest that immune suppression induced by the human immunodeficiency virus increases MCC risk. PMID- 11853801 TI - Tackle infectious disease to help the poor, says WHO. PMID- 11853804 TI - New understanding into the mechanics of migraine. PMID- 11853805 TI - Mixed benefits of hormone replacement therapy. PMID- 11853806 TI - Early weight gain patterns may affect adult blood pressure. PMID- 11853807 TI - Do violent movies make violent children? PMID- 11853808 TI - Assistance for Serbs and Roma from Kosovo. PMID- 11853809 TI - European union continues negotiations on tobacco control. PMID- 11853810 TI - South African government procrastinates over policy on HIV drug. PMID- 11853811 TI - Catalan authorities investigate insurance fraud plot by doctors and lawyers. PMID- 11853812 TI - Ireland prepares for referendum on abortion. PMID- 11853815 TI - Germany's parliament approves imports of embryonic stem cells. PMID- 11853816 TI - Viral encephalitis: familiar infections and emerging pathogens. AB - Significant advances have been made in our understanding of the natural history and pathogenesis of viral encephalitides. The development of PCR has greatly increased our ability to diagnose viral infections of the central nervous system, particularly for herpes and enteroviral infections. Advancing knowledge has led to the recognition that some encephalitides can be reliably prevented by vaccination (eg, Japanese encephalitis and rabies). For other pathogens such as the arboviruses, the focus has been on prevention by vector control. Finally, effective therapy has been established for a very limited number of viral infections (eg, acyclovir for herpes simplex encephalitis). Other potentially useful treatments, such as pleconaril for enteroviral meningoencephalitis are under clinical evaluation. We review current understanding of viral encephalitides with particular reference to emerging viral infections and the availability of existing treatment regimens. PMID- 11853818 TI - Generation of allocation sequences in randomised trials: chance, not choice. AB - The randomised controlled trial sets the gold standard of clinical research. However, randomisation persists as perhaps the least-understood aspect of a trial. Moreover, anything short of proper randomisation courts selection and confounding biases. Researchers should spurn all systematic, non-random methods of allocation. Trial participants should be assigned to comparison groups based on a random process. Simple (unrestricted) randomisation, analogous to repeated fair coin-tossing, is the most basic of sequence generation approaches. Furthermore, no other approach, irrespective of its complexity and sophistication, surpasses simple randomisation for prevention of bias. Investigators should, therefore, use this method more often than they do, and readers should expect and accept disparities in group sizes. Several other complicated restricted randomisation procedures limit the likelihood of undesirable sample size imbalances in the intervention groups. The most frequently used restricted sequence generation procedure is blocked randomisation. If this method is used, investigators should randomly vary the block sizes and use larger block sizes, particularly in an unblinded trial. Other restricted procedures, such as urn randomisation, combine beneficial attributes of simple and restricted randomisation by preserving most of the unpredictability while achieving some balance. The effectiveness of stratified randomisation depends on use of a restricted randomisation approach to balance the allocation sequences for each stratum. Generation of a proper randomisation sequence takes little time and effort but affords big rewards in scientific accuracy and credibility. Investigators should devote appropriate resources to the generation of properly randomised trials and reporting their methods clearly. PMID- 11853819 TI - Medical professionalism in the new millennium: a physicians' charter. PMID- 11853820 TI - Early anticoagulation in acute ischaemic stroke. PMID- 11853821 TI - Early anticoagulation in acute ischaemic stroke. PMID- 11853823 TI - Antimalarial agents in pregnancy. PMID- 11853824 TI - Soluble adhesion molecules and coronary heart disease. PMID- 11853825 TI - Soluble adhesion molecules and coronary heart disease. PMID- 11853826 TI - Soluble adhesion molecules and coronary heart disease. PMID- 11853828 TI - Pseudomonas aeruginosa cross-infection in cystic fibrosis. PMID- 11853830 TI - In-vivo dedifferentiation of keratinocytes to epidermal stem cells. PMID- 11853831 TI - In-vivo dedifferentiation of keratinocytes to epidermal stem cells. PMID- 11853832 TI - In-vivo dedifferentiation of keratinocytes to epidermal stem cells. PMID- 11853834 TI - Female genital mutilation: why are we so radical? PMID- 11853835 TI - Professionalism of doctors in the UK. PMID- 11853836 TI - Professionalism of doctors in the UK. PMID- 11853837 TI - Professionalism of doctors in the UK. PMID- 11853838 TI - Darwin the philosopher? PMID- 11853840 TI - Risk of deliberately induced anthrax outbreak. PMID- 11853841 TI - NHS: and the tap goes drip, drip, drip. PMID- 11853842 TI - Monitored anaesthesia in elderly ophthalmic patients. PMID- 11853850 TI - Walking to the ward. PMID- 11853852 TI - Cognitive deficit and development of motor impairment in a mouse model of Niemann Pick type C disease. AB - Niemann-Pick disease type C (NPC) is a fatal, autosomal recessive lipidosis characterized by a unique error in cellular trafficking of cholesterol. In the disease, unesterified cholesterol as well as sphingolipids accumulate in the late endosomes/lysosomes due to mutations in either of two recently isolated genes, NPC1 or NPC2. A metabolic and neurological disorder reminiscent of human NPC disease has been described in Balb/C mice, and it was recently shown that the mutation in the NPC mice resides in the orthologous murine Npc1 gene. Here we have followed the growth rate and applied behavioural methods in order to establish the onset and development of the major symptoms in the NPC mouse model. Wild type and NPC mice were studied during 28-59 days of age. Both male and female NPC mice displayed retarded growth at the age between 25 and 35 days. At the age of 35-45 days the weight was similar to controls and thereafter very rapidly decreased. The battery of coordination tests (vertical screen, beam balancing, coat hanger and rotating rod) established motor impairment of the NPC mice already at the age of 28-42 days, well before the onset of visually detectable ataxia. Decreased exploratory activity and lack of habituation was revealed in the NPC mice by open field test. The diseased mice were unable to learn and remember the location of the hidden escape platform in spatial water maze task suggesting cognitive impairment. In several tests the male NPC mice were more affected than the females. The present study represents the first behavioural analysis of the NPC mice. The battery of behavioural tests employed here should be valuable in the assessment of effective approaches to treat NPC, for which no preventive or curative measures have so far been established. PMID- 11853853 TI - The role of climbing and parallel fibers inputs to cerebellar cortex in navigation. AB - DA-HAN rats with partial or total lesion of climbing (CF) and parallel fibers (PF) inputs of the cerebellum were tested in a water task. Two different protocols were used, requiring to find either a non-visible or a visible platform. These two protocols were, respectively, designed to evaluate visuo motor guidance (visible platform) and navigation (non-visible platform). Both groups of lesioned rats presented a deficit in the non-visible platform task but not in the visible platform one. The protocol of navigation we used was a fixed start-fixed arrival procedure. Totally lesioned animals were unable to learn to orient their body toward the non-visible platform and adopted instead a circling behavior. Our results suggest a role of cerebellar inputs (climbing (CF) and PF) in navigation. PMID- 11853854 TI - Role of the anterodorsal and anteroventral nuclei of the thalamus in spatial memory in the rat. AB - This study tests the hypothesis that the anterior thalamic nuclei play a significant role in spatial learning and memory. Adult, male Sprague-Dawley rats with bilateral ibotenic acid lesions of the anterior thalamus were tested for 5 days in a repeated acquisition water maze task. Compared with Controls, rats with nearly complete lesions of both anterodorsal (AD) and anteroventral (AV) thalamic nuclei (AD/AV) were only mildly impaired in their spatial learning and memory. Larger lesions that extended into the anteromedial (AM) thalamic nucleus (AD/AV+) caused a more severe impairment and complete lesions of all three anterior nuclei (AD/AV/AM) resulted in even greater impairment that extended to all aspects of the task. In probe trials, only the Control animals had a preference for the correct quadrant. Approximately one-half of the rats were tested for a second week to determine if the impaired groups would benefit from further training. AD/AV/AM rats showed little improvement, but the other groups all improved significantly in all aspects of the task except the probe trial. Together, these data indicate that the anterior thalamic nuclei contribute to spatial learning and memory, but neither AV nor AD independently plays a dominant role. PMID- 11853855 TI - Impaired motor activity and motor learning function in rat with middle cerebral artery occlusion. AB - The poor quality of life after a stroke is largely attributed to deficits in cognitive-motor functioning. The goals of this study were to detect if damaged motor learning function were attributed to motor deficits in rats following a transient middle cerebral artery (MCA) occlusion. Stroke was induced by a 2-h occlusion of the MCA using an intraluminal filament. Motor functions were evaluated from 5 up to 28 days after reperfusion in ischemic and control rats. Motor function was detected by a series of motor tests (runway traversing and beam balancing, as well as foot fault placing, parallel bar crossing, rope and ladder climbing), and motor learning behavior was determined by analyzing the rate of improvement of impaired function during performance of the motor tasks. Significant (P<0.001) motor deficits were detected in the stroke group (n=10) while performing motor tasks that involve extensive coordination, in comparison to the controls (n=12). Although motor behavior was improved with repeated behavior testing, unparalleled rate of improvement of motor performance on rope and ladder climbing tests was found between the two groups, suggesting an impaired motor learning function. Brain tissue damage was detected in the ischemic animals 28 days after surgery, demonstrated by 40% infarct volume of contralateral hemisphere. Both motor learning and motor function were impaired in ischemic rats. The motor tests used in this study are sensitive, semi quantitative, and reproducible measurements of functional impairment in rats following an ischemic stroke. PMID- 11853856 TI - 5-HT1A agonist/antagonist modification of cocaine stimulant effects: implications for cocaine mechanisms. AB - The 5-HT1A receptor site has been demonstrated to be an important pharmacological target in the modulation of unconditioned behavioral effects induced by cocaine. In this study, separate groups of rats (n=7) received a series of the 5-HT1A agonist treatments, 8-OHDPAT (0.2,0.4 mg/kg) in combination with saline or cocaine (10 mg/kg). Using a crossover design, the treatments were subsequently switched to the 5-HT1A antagonist, WAY 100635 (0.4,0.8 mg/kg) and then, switched back again to 8-OHDPAT (0.2,0.4 mg/kg). When the 8-OHDPAT was given in combination with cocaine, locomotion was substantially enhanced but when the treatment was switched to WAY 100635, the cocaine induced locomotion was suppressed. Neither the 8-OHDPAT or WAY 100635 given with saline affected locomotion as compared to saline treated animals. These findings indicated a reciprocal facilitatory/inhibitory influence of 5-HT1A agonists/antagonists upon cocaine induced locomotion. The 8-OHDPAT treatments, however, did not enhance all cocaine behavioral responses. Initially, 8-OHDPAT suppressed cocaine induced rearing and central zone entry, but with repeated treatments, these response suppression effects subsided. As a consequence, the facilitative influence of 8 OHDPAT upon cocaine induced locomotion could not be attributed to response redistribution effects. While WAY 100635 markedly reduced cocaine induced locomotion and rearing to nearly saline response levels, the same WAY 100635 treatments did not modify locomotor stimulant effects induced by caffeine (10 mg/kg). In that caffeine stimulant effects are not directly linked to serotonergic mechanisms, the absence of an influence of WAY 100635 upon caffeine induced locomotor stimulation lent further support to the proposition that the 5 HT1A receptor site contributes to locomotor behavior in situations where the serotonergic system is pharmacologically activated by drugs such as cocaine. These findings point to a potential role for 5-HT1A antagonists in treatment of cocaine abuse. PMID- 11853857 TI - PET studies on the memory processing of word pairs in bilingual Finnish-English subjects. AB - This study examined the fundamental question whether verbal memory processing in two unrelated languages is mediated by a common neural system or by distinct cortical areas. Ten right-handed, male Finnish--English adult late bilinguals who had acquired the second language after the age of 10 were scanned whilst either encoding/retrieving word pairs in their mother tongue (Finnish) or in a foreign language (English). Within each language, subjects had to encode and retrieve four sets of 12 visually presented paired word associates which were not semantically related. Two sets consisted of highly imageable words (e.g. monkey table; koira-lasi) and the other two sets of abstract word pairs (e.g. freedom moral; uhka-suure). Presentation of pseudowords served as a reference condition. An emission scan was recorded after each intravenous administration of O-15 water. Encoding was associated with prefrontal and hippocampal activation. During memory retrieval, precuneus showed a consistent activation in both languages and for both highly imageable and abstract words. Although the brain mechanisms of the two languages share common components, differential activations were found in Broca's area and in the cerebellum as well as in the angular/supramarginal gyri according to the language used. PMID- 11853858 TI - Dynamics of hand preference in 4 year-old children. AB - Hand preference in groups of 4 year-old children towards either end of the skill continuum (as determined by the Movement ABC test) was explored by means of a catching task in which the 'direction of approach of the ball' was used as a control parameter within a dynamical systems framework. In Condition 1, the ball direction was systematically scaled in degrees from the right side of the subject to the left and vice versa. In Condition 2, the spatial location of the ball direction was varied randomly, rather than systematically, over the same range as for Condition 1. In both conditions, the two groups of children were shown to switch between hands (from left to right and vice versa) at well-defined positions on the scale. The different scale values obtained when ball direction was scaled from left to right compared with right to left were used to define hysteresis areas. These areas were more extensive and located more to the left of the midline for the group towards the less skilled end of the continuum than for those towards the more skilled end. Additionally, the less-skilled children were shown to use the right hand more to catch the balls delivered to the left side than did the more skilled children. In Condition 3, objects were placed on the table of the ball-delivery apparatus in order to make the catching action more complicated. The less skilled group were shown to use the right hand more than the left hand in all catching positions whereas the more skilled group showed an equal usage of both catching regimes. PMID- 11853859 TI - Variability factors in the expression of stress-induced behavioural sensitisation. AB - Altered behavioural and physiological responsivity following a short session of foot shocks in the rat has proven to be a stable and clinically relevant model of stress-induced sensitisation. However, a number of key factors influencing effect size or direction have not previously been reported. Rats underwent a single, 15 min session of foot shocks and were exposed to a variety of novel stressful challenges 1 or 2 weeks later. Sensitised behavioural responses (increased immobility) in preshocked rats remained present over 3 days of repeated exposure to noise stress. In mild novel challenges (open field, empty cage), behavioural sensitisation and defecation was most clearly expressed at the beginning of the dark phase (evening). Higher-arousal challenges (prod, noise) caused increased behavioural inhibition in preshocked rats at all three time points (morning, afternoon, evening). Female preshocked rats showed a different pattern of behavioural and defecation sensitisation than preshocked males. The robustness of the model makes it suitable for further investigations into the mechanisms and vulnerability factors involved in the long-term consequences of stress. PMID- 11853860 TI - Humans with hippocampus damage display severe spatial memory impairments in a virtual Morris water task. AB - For nonhumans, it has been shown that the hippocampus (HPC) is critical for spatial memory. We tested patients with unilateral HPC resections on a virtual analogue of a classic spatial task to assess HPC functioning in nonhumans: the Morris water task. We found that when humans are required to use spatial cues to navigate to a hidden escape platform in a pool, patients with HPC resections display severe impairments in spatial navigation relative to age-matched controls and age-matched patients who have had extra-HPC resections. This effect occurred for every patient tested and was evident regardless of side of surgery. Hence, it is apparent across species and irrespective of which hemisphere is damaged that the human HPC is critical for spatial/relational memory. PMID- 11853861 TI - Estrogen's effects on activity, anxiety, and fear in two mouse strains. AB - Estrogen has effects on activity levels and emotional reactivity in both humans and rats. In a recent study conducted in ovariectomized (OVX) C57BL/6 (C57) mice we found that treatment with estradiol benzoate (EB) increased anxiety, fear learning, and running wheel activity relative to vehicle control (Veh). The present study was conducted to examine the stability of these findings across mouse strains (C57 and Swiss-Webster; SW), to get a better sense of the magnitude of the anxiety response by reducing baseline anxiety levels, and to discover if EB affects activity levels in a safe environment other than the home-cage running wheel. Mice of both strains treated with EB (s.c. implant, 25 microg in sesame oil, which enters the body over 5 weeks) were more anxious than Veh animals in the open field, elevated plus, and dark-light transition tests. SW animals were less anxious than C57 in the elevated plus. EB-treated animals of both strains were more active in the running wheel than Veh animals, and more active in the test of spontaneous activity in the home cage. EB-treatment also increased fear learning in a step-down avoidance task. EB appears to have a consistent but moderate effect in elevating anxiety and in increasing fear learning in two strains of mice. It is also involved in increasing activity in two different types of locomotion in the safer home cage. We conclude that these results of increased anxiety/fear and increased activity are suggestive of a general increase in arousal, with both sets of responses increasing the likelihood of reproductive behaviors occurring only when the environment predicts success. PMID- 11853862 TI - Attentional set-shifting in mice: modification of a rat paradigm, and evidence for strain-dependent variation. AB - Increasingly precise molecular genetic tools are available to study in mice the cellular mechanisms underlying complex brain functions, but the behavioural paradigms to assess these functions often lack the required specificity. In this study, an attentional set-shifting paradigm to assess medial frontal cortex functions in rats was modified for use in mice and variation between two relevant mouse strains assessed. Male 129/SvEv and C57BL/6J mice and their F1 intercross (n=8 per genotype) were trained to dig in bowls for a food reward. On four consecutive days, mice performed a series of discriminations to criterion (six consecutive correct choices) between pairs of food bowls that differed along two dimensions (odour, digging medium), including a reversal, an intra-dimensional shift, and an extra-dimensional shift. Mice from the 129 strain performed significantly better than C57 mice in the initial acquisition of a simple discrimination and in the final extra-dimensional shift test, with no difference in the reversal and intra-dimensional shift. Performance of the F1 mice was intermediate or similar to that of the 129 mice. These results indicate a selective difference between these two strains in attentional selection processes that have been shown in humans, monkeys and rats to be mediated by prefrontal cortex. PMID- 11853863 TI - Behavioral responses to ingestion of different sources of fat. Involvement of serotonin? AB - In previous experiments, we described a decrease in extracellular hypothalamic serotonin, 5-hydroxy-tryptamine (5-HT), in response to ingestion of a lard meal. This effect was related to the low satiating potency of lard as compared with other nutrients. Since the composition in fatty acids might be important for these effects, the purpose of this study was to analyze the neurochemical and behavioral responses to ingestion of different sources of fat. Unique meals of three margarines used in human feeding were compared with a meal of lard with regard to their effects on hypothalamic 5-HT, satiety, anxiety-like behavior and sucrose consumption. A vegetable margarine with a high content in saturated fatty acids also decreased hypothalamic 5-HT while ingestion of a sunflower oil and an olive oil enriched margarines, both high in polyunsaturated fatty acids, did not affect significantly 5-HT levels. However, these two last ones were not the most satiating. The olive oil margarine induced a tendency to an anxiety-like behavior while lard increased sucrose consumption. Thus, ingestion of fats may alter specifically behavioral responses. The involvement of 5-HT is likely in the case of lard ingestion but probably not for the other sources of fat. PMID- 11853864 TI - Perception of the corridor illusion by baboons (Papio papio). AB - The corridor illusion was assessed in four baboons (Papio papio) by way of judgmental task implying a comparison between the size of two figures presented on various backgrounds. Findings demonstrate that the baboons are sensitive to the corridor illusion. PMID- 11853865 TI - Exogenous endothelin-1 improves microvascular oxygen balance during focal cerebral ischemia in the rat. AB - We tested the hypothesis that endothelin-1 (ET-1), a cerebrovasoconstrictive peptide, would alter microvascular oxygen balance during focal cerebral ischemia. In this study, male Wistar rats were placed in control (n=9) and ET-1-treated (n=9) groups. Cortical ischemia was induced by middle cerebral artery (MCA) occlusion in isoflurane (1.4%) anesthetized rats. Forty minutes after MCA occlusion, 10(-7) M ET-1 or saline was applied to the ischemic cortex (IC) for a period of 20 min; the fluid was changed every 5 min. After 1 h of ischemia, regional cerebral blood flow (rCBF) was determined using a 14C-iodoantipyrine autoradiographic technique. Regional arterial and venous oxygen saturation were determined microspectrophotometrically. The cerebral blood flow (45% control, 45% ET-l) and oxygen consumption (24% control, 44% ET-1) of the IC were significantly lower than the contralateral cortex. ET-1 of 10(-7) M did not cause a statistically significant alteration in regional cerebral blood flow or oxygen consumption of the IC, but did increase the average venous O(2) saturation of the IC from 50 +/- 1% to 55 +/- 2% (p<0.04). A significant (p<0.05) increase was observed in O(2) supply/consumption ratio in the ET-1-treated IC (2.79 +/- 0.26 ml O(2)/min/100 g in ET-1-treated IC vs. 2.41 +/- 0.12 ml O(2)/min/100 g in the control IC) compared to the control IC. ET-1 also significantly lowered the frequency of small veins with less than 50% O(2) saturation in the IC (39 out of 70 veins in IC vs. 17 out of 70 veins in ET-1-treated IC). Thus, the exogenous application of 10(-7) M ET-1 improved microvascular oxygen supply/consumption balance during focal cerebral ischemia. PMID- 11853866 TI - Inhibition of mu and delta opioid receptor ligand binding by the peptide aldehyde protease inhibitor, leupeptin. AB - We reported recently that the ubiquitin-proteasome pathway is involved in agonist induced down regulation of mu and delta opioid receptors [J. Biol. Chem. 276 (2001) 12345]. While evaluating the effects of various protease inhibitors on agonist-induced opioid receptor down regulation, we observed that while the peptide aldehyde, leupeptin (acetyl-L-Leucyl-L-Leucyl-L-Arginal), did not affect agonist-induced down regulation, leupeptin at submillimolar concentrations directly inhibited radioligand binding to opioid receptors. In this study, the inhibitory activity of leupeptin on radioligand binding was characterized utilizing human embryonic kidney (HEK) 293 cell lines expressing transfected mu, delta, or kappa opioid receptors. The rank order of potency for leupeptin inhibition of [3H]bremazocine binding to opioid receptors was mu > delta > kappa. In contrast to the effect of leupeptin, the peptide aldehyde proteasome inhibitor, MG 132 (carbobenzoxy-L-Leucyl-L-Leucyl-L-Leucinal), had significantly less effect on bremazocine binding to mu, delta, or kappa opioid receptors. We propose that leupeptin inhibits ligand binding by reacting reversibly with essential sulfhydryl groups that are necessary for high-affinity ligand/receptor interactions. PMID- 11853867 TI - Tachykinin-stimulated small bowel myoelectric pattern: sensitization by NO inhibition, reversal by neurokinin receptor blockade. AB - Tachykinins stimulate motility whereas NO inhibits motility in the gastrointestinal tract. AIM: To investigate if inhibition of NO production sensitizes myoelectric activity to subthreshold doses of tachykinins in the small intestine of awake rats. METHODS: Rats were supplied with a venous catheter and bipolar electrodes at 5, 15 and 25 cm distal to pylorus for electromyography of small intestine. The motor responses were evaluated using pattern recognition. Substance P and neurokinin A dose-dependently stimulated gut motility, with neurokinin A being more potent than substance P. Therefore, neurokinin A was chosen and administered under baseline conditions and 45-60 min after N(omega) nitro-L-arginine (L-NNA) 1 mg kg(-1), with or without pretreatment with L arginine 300 mg kg(-1). In addition, myoelectric activity effects of neurokinin A in conjunction with L-NNA were studied before and after administration of the tachykinin receptor antagonists, SR140333 (NK1), SR48968 (NK2) and SR142801 (NK3), each at 2.5 mg kg(-1). RESULTS: Dose-finding studies verified 10 pmol kg( 1) min(-1) to be the threshold dose at which NKA caused phase II-like activity in a low percentage of experiments (12%, n=41). This dose was therefore used in combination with L-NNA for sensitization experiments of gut myoelectric activity. In experiments where NKA-induced no response, pretreatment with L-NNA led to phase II-like activity in 9 of 18 (50%, p<0.05) experiments. Co-administration of SR140333 and SR48968 abolished this effect. CONCLUSION: NO counteracts the stimulatory effect of tachykinins on small bowel myoelectric activity in the rat. Inhibition of the L-arginine/NO pathway sensitizes the gut to tachykinin stimulated motor activity. PMID- 11853868 TI - Postnatal profile of plasma leptin concentrations in male and female rats: relation with the maturation of the pituitary-gonadal axis. AB - Leptin, the ob gene product, has been implicated in the initiation of puberty in mice and humans. However, it is not yet clear whether leptin also plays a pivotal role in promoting sexual maturation in rats. Based on the assumption that circulating leptin levels would increase during the peripubertal period if this hormone triggers puberty, we examined the developmental profile of plasma leptin from neonatal (day 1) through adult (day 85) age in both male and female rats and simultaneously monitored several important indices of pituitary-gonadal function. A significant elevation of plasma leptin during the peripubertal period was not observed in either male or female rats. Although this finding may not support leptin as a humoral factor triggering puberty, we observed a rise in leptin in both sexes from the second postnatal week, which clearly preceded the first significant elevations of luteinizing hormone and gonadal steroids. Therefore, it is still possible that leptin may play a role in promoting sexual maturation in rats of both sexes already from an early postnatal period. It seems that the role for leptin in sexual development is permissive, but not decisive, in rats. PMID- 11853869 TI - Cardiovascular actions of central neuromedin U in conscious rats. AB - Neuromedin U (NMU) is a brain-gut peptide, which peripherally stimulates smooth muscle, increases of blood pressure, alters ion transport in the gut, controls local blood flow, and regulates adrenocortical function. Although intracerebroventricular (i.c.v.) administration of NMU is known to decrease food intake and body weight, little is known about its effect on other physiological functions. We examined the effects of i.c.v. administration of NMU on mean arterial pressure (MAP), heart rate (HR), and plasma norepinephrine in conscious rats. Neuromedin U (0.05 and 0.5 nmol) provoked an increase in MAP (93.8 +/- 0.5 to 123.5 +/- 1.7 and 94.7 +/- 0.8 to 132.7 +/- 3.0 mm Hg, respectively) and HR (334.9 +/- 6.0 to 494.1 +/- 6.9 and 346.3 +/- 3.3 to 475.1 +/- 8.9 beats/min, respectively). In contrast, plasma norepinephrine increased only with a high dose of neuromedin U. Intravenously administered NMU (0.5 nmol) elicited a small and short lasting increase in MAP, compared to that by i.c.v. NMU. These results indicate that central neuromedin U regulates sympathetic nervous system activity and affects cardiovascular function. PMID- 11853870 TI - Transendothelial migration of leukocytes and signalling mechanisms in response to the neuropeptide secretoneurin. AB - Secretoneurin (SN), a newly discovered neuropeptide, may be implicated in inflammatory responses as it was shown to modulate leukocyte, endothelial and mesenchymal cell functions. Neutrophils placed above pulmonary arterial or venous endothelial monolayers migrated through this cellular barrier in response to apical or basal stimulation with SN in a dose-dependent manner. At optimal concentrations of 10(-6) to 10(-8) M, SN was nearly equally effective in stimulating neutrophil transmigration as was tumor necrosis factor-alpha at 10 ng/ml or a chemotactic gradient of formyl-Met-Leu-Phe (10(-8) M). Stimulation of transendothelial migration appears to be specific, since a trypsin digest of SN was ineffective and excess concentrations of anti-SN antibodies completely abolished the effect. Inhibition of cyclooxygenase or nitric oxide synthase did not affect the action of SN. Preincubation of endothelial cells with pertussistoxin (PTx) or choleratoxin (CTx), and the presence of staurosporine significantly inhibited transmigration, suggesting that SN uses a signalling pathway that is coupled to G-proteins and protein kinase C in endothelium. Moreover, SN treatment resulted in transient elevation of cytoplasmatic calcium concentration in endothelial cells. These data support the hypothesis that SN might contribute to neurogenic inflammation in vivo and reveal signalling mechanisms of SN in endothelial cells. PMID- 11853872 TI - Pharmacological characterization of cholecystokinin receptors mediating contraction of human gallbladder and ascending colon. AB - Cholecystokinin (CCK) produces contractions of gallbladder and colon in a number of different species. Although the effects of CCK on the human gallbladder are relatively well documented, the CCK receptors in the human colon have not been clearly characterised. Therefore, in this study, the CCK receptors in the human gallbladder and colon were compared using pharmacological techniques. Contraction of specimens of the human tissue was measured using in vitro organ bath bioassay. The effect of selective concentrations of CCK(1) and CCK(2) receptor antagonists (L-364,718 and JB93182, respectively) was determined on agonist concentration effect (E/[A]) curves obtained by cumulative dosing with sulphated CCK. The CCK(1) antagonist L-364,718 produced a rightward shift of the CCK-8S [E/[A] curve in the human gallbladder (pA(2)=9.15 +/- 0.26) and ascending colon (pA(2)=9.20 +/ .33). In both tissues, the CCK(2) receptor antagonist, JB93182, had no effect on the CCK E/[A] curves. In addition, in the colon, pentagastrin responses were inhibited by L-364,718 but unaffected by JB93182. These data indicate that the CCK-induced contraction of the human colon and gallbladder smooth muscle is mediated solely through the CCK(1) receptor subtype, and the antagonist affinity estimates are consistent with those previously obtained in experiments on animal tissue. PMID- 11853871 TI - AT(1) receptor blockade with losartan during gestation in Wistar rats leads to an increase in thirst and sodium appetite in their adult female offspring. AB - We studied the effects of angiotensin II receptor blockade with losartan on thirst and sodium appetite in pregnant Wistar rats and on their adult female offspring. During maternal adaptation to pregnancy, average daily total water intake increased by 63% (P<0.01); NaCl intake by 214% (P<0.001). These changes were not blocked by daily s.c. injections of losartan (50 mg/kg bw i.p.) from gestation day (GD) 2 until GD 19 which implied that maternal AT(1) receptors were not involved in the up regulation of thirst and sodium appetite during pregnancy. Losartan blockade during gestation led to a significant and continued increase in thirst and sodium appetite in the adult female offspring. Daily water intakes were greater in the losartan (LO) group than in the vehicle-injected control group (CO), leading to a total water intake of 1114 +/- 80.6 ml/kg bw compared with 738 +/- 56.7 ml/kg bw (P<0.05) during the 8-day period of observation. Daily sodium intakes were usually 2-3 times greater in the LO group compared with the CO group, amounting to a final cumulative intake of 232 +/- 33 mmol/kg bw compared with 93.8 +/- 16.5 mmol/kg bw (P<0.05) in 8 days. These elevated sodium and water intakes were nearly counterbalanced by the increased renal excretion of water and sodium by fully functional kidneys that were not injured by the drug. Body weights were 10% lower in the LO group at the start but remained unchanged relative to the CO group during the entire 8-day period of observation. Plasma electrolytes, blood hematocrit and carotid MABP in the LO group did not differ from the CO group. PMID- 11853874 TI - Multiple roles of the invariant chain in MHC class II function. PMID- 11853873 TI - Activation of extracellular signal regulated protein kinase by neuropeptide Y and pancreatic polypeptide in CHO cells expressing the NPY Y(1), Y(2), Y(4) and Y(5) receptor subtypes. AB - Neuropeptide Y (NPY), 36-amino acid amidated peptide expressed in central and peripheral neurons, regulates a variety of physiological activities, including food intake, energy expenditure, vasoconstriction, anxiolysis, nociception and ethanol consumption. NPY binds to a family of G-protein coupled receptors whose activation results in inhibition of adenylyl cyclase activity. To more fully characterize the signal transduction pathways utilized by the NPY receptor subtypes, the pathways leading to phosphorylation of the extracellular signal regulated protein kinases 1 and 2 (ERK) have been compared in CHO cells expressing each of the four cloned human NPY receptor subtypes, Y(1), Y(2), Y(4) and Y(5). NPY Y(1), Y(2), Y(4) and Y(5) receptor-mediated ERK phosphorylation was blocked by pertussis toxin (PTX) exposure, indicating that all four receptors are coupled to inhibitory G(i/o) proteins. Exposure to the protein kinase C (PKC) inhibitor GF109203X diminished Y(1), Y(2) and Y(4) receptor-mediated ERK phosphorylation but completely blocked Y(5) receptor-mediated ERK phosphorylation. Additionally, Y(5) receptor-mediated ERK phosphorylation was inhibited by the phosphatidylinositol 3-kinase inhibitors LY294002 and wortmannin to a greater extent than was Y(1)-mediated ERK phosphorylation. These results demonstrate that in CHO cells, the Y(5) receptor and the Y(1), Y(2) and Y(4) receptors utilize different pathways to activate ERK. PMID- 11853875 TI - The involvement of NF-kappaB in the constitutive overexpression of cyclooxygenase 2 in cyclooxygenase-1 null cells. AB - We recently reported that there was enhanced cyclooxygenase (COX)-2 expression and prostaglandin E(2) biosynthesis in COX-1 deficient (COX-1(-/-)) cells. We also observed that the growth of COX-1(-/-) cells was significantly retarded compared to wild-type (WT) and COX-2 deficient (COX-2(-/-)) cells. In this study, COX-2 expression and its promoter activity were compared in immortalized, nontransformed fibroblasts from WT, COX-1(-/-) or COX-2(-/-) mice in the context of the role of COX-2 as a growth regulator. When compared with WT cells expressing both COX isoenzymes, constitutive COX-2 protein and promoter activity were significantly higher in COX-1(-/-) cells as determined by Western blotting and luciferase assays using a 5'-flanking promoter construct of the murine COX-2 gene. The luciferase assay using a series of luciferase-linked COX-2 promoter deletions transfected into COX-1(-/-) cells indicated that a region involving NF kappaB plays a significant role in regulating constitutive COX-2 expression. Data from electrophoretic mobility shift assays showed that COX-1(-/-) cells contained higher levels of activated NF-kappaB than either WT or COX-2(-/-) cells. Furthermore, COX-2 promoter activity was significantly inhibited by the oligonucleotides (ODNs) containing the NF-kappaB element (NF-kappaB decoy ODNs) but not by the scrambled control ODNs, as examined by the luciferase assay. These findings indicate that constitutive COX-2 promoter activity and protein expression are enhanced in COX-1(-/-) fibroblasts and that signaling via the NF kappaB pathway is involved in the transcriptional control of constitutive COX-2 expression. PMID- 11853876 TI - The alpha5beta1 integrin selectively enhances epidermal growth factor signaling to the phosphatidylinositol-3-kinase/Akt pathway in intestinal epithelial cells. AB - We have investigated EGF-driven signaling processes in rat intestinal epithelial cell lines that overexpress either the alpha5beta1 integrin or the alpha2beta1 integrin. Both cell types display efficient activation of Erk/MAP kinase, but only the alpha5beta1 expressing cells display a strong activation of Akt. A complex is formed between activated EGFR and alpha5beta1, but not with alpha2beta1; this complex also contains ErbB3 and p85. Thus alpha5beta1 can support efficient activation of both the Erk and the phosphatidylinositol-3 kinase/Akt branches of the EGFR signaling cascade, whereas alpha2beta1 can support only the Erk branch. PMID- 11853877 TI - Novobiocin is a novel inducer of CD38 on cells of the myelomonocytic lineage. AB - KG-1a, HL-60 and U-937 cells, which represent different stages of myelopoiesis, showed growth retardation in response to the coumarin antibiotic novobiocin. Novobiocin was found to increase CD38 expression (in all three cell lines) and to induce differentiation along the monocytic path in HL-60 and U-937 cells but not in KG-1a cells. The increase in surface expression of CD38 was matched by NAD glycohydrolase activity and by increases in the level of specific mRNA, indicating that the gene product is active and that regulation occurs at the level of transcription or mRNA stability. Of the three cell lines tested, only the early KG-1a expressed N-cadherin, a member of Ca(2+)-dependent adhesion molecules involved in embryonic differentiation processes. In contrast to CD38, N cadherin was slightly down-regulated pointing to a specific role of novobiocin in gene regulation. PMID- 11853878 TI - Characterization of ubiquilin 1, an mTOR-interacting protein. AB - The mTOR protein kinase is known to control cell cycle progression and cell growth through regulation of translation, transcription, membrane traffic and protein degradation. Known interactions of mTOR do not account for the multiple functions of this protein. Using a non-catalytic segment of mTOR (1-670) as bait in a yeast two-hybrid screen for interacting proteins, ubiquilin 1 (NM013438) was identified. Ubiquilin 1 is a member of a phylogenetically conserved gene family of unknown function, characterized by an N-terminal ubiquitin-like (Ubq) domain, a C-terminal ubiquitin associated (Uba) domain and a central region containing numerous NPXvar phi motifs (X, any; phi, hydrophobic amino acid). GST-ubiquilin 1 binds specifically to FLAG-mTOR (residues 1-670) in mammalian cells; residues 570 670 of mTOR and 226-323 of ubiquilin 1 are required for this interaction. Both mTOR and ubiquilin immunoreactivity appear as fine speckles throughout the cytoplasm; significant colocalization with cytoskeletal elements, early endosomes or proteasomes is not observed. As assessed by cell fractionation, mTOR is predominantly associated with low density membranes, along with 10% of ubiquilin 1. Ubiquilin 1 is a rapamycin-insensitive phosphoprotein. Overexpression of ubiquilin 1 does not alter the kinase activity of cotransfected mTOR or the phosphorylation of the mTOR target, p70 S6 kinase, in the presence or absence of rapamycin. Our data suggest that we have identified a novel mTOR interactor, ubiquilin 1. The biological significance of this, presumably membrane based, interaction, requires further study. PMID- 11853879 TI - Regulation of ornithine decarboxylase in B16 mouse melanoma cells: synergistic activation of melanogenesis by alphaMSH and ornithine decarboxylase inhibition. AB - Ornithine decarboxylase (ODC) is the rate-limiting enzyme in the biosynthesis of polyamines, a family of cationic compounds required for optimal cell proliferation and differentiation. Within mammalian melanocytes, the expression of genes regulating cell growth and/or differentiation can be controlled by alpha melanocyte-stimulating hormone (alphaMSH) and other melanogenesis modulating agents. In the B16 mouse melanoma model, alphaMSH stimulates melanogenesis by upmodulation of tyrosinase (tyr) activity, whereas the phorbol ester 12-O tetradecanoylphorbol 13-acetate (TPA) inhibits melanin synthesis. Therefore, we analyzed the regulation of ODC by these agents, as related to changes in the melanogenic pathway. Treatment of B16 cells with TPA or alphaMSH rapidly stimulated ODC activity. The effect was stronger for TPA and appeared mainly posttranslational. Irreversible inhibition of ODC with the active site-directed inhibitor alpha-difluoromethylornithine (DFMO) did not block TPA-mediated inhibition of tyr. Conversely, prolonged treatment of B16 cells with DFMO stimulated tyr activity by a posttranslational mechanism, probably requiring polyamine depletion. Combination treatment with alphaMSH and DFMO synergistically activated tyr. Therefore, ODC induction is not involved in the melanogenic response of B16 cells to alphaMSH. Rather, increased intracellular concentrations of polyamines following ODC induction might constitute a feedback mechanism to limit melanogenesis activation by alphaMSH. PMID- 11853880 TI - Tumor necrosis factor-alpha exerts interleukin-6-dependent and -independent effects on cultured skeletal muscle cells. AB - In vivo studies have shown that cancer-associated skeletal muscle wasting (cachexia) is mediated by two cytokines, tumor necrosis factor-alpha (TNF) and interleukin-6 (IL-6). It has been unclear from these studies whether TNF exerts direct effects on skeletal muscle and/or whether these effects are mediated via IL-6. Previous studies from our laboratory have shown that TNF induces IL-6 mRNA expression in cultured skeletal muscle cells. To further investigate the relationship between TNF and IL-6, the effects of TNF and IL-6 on protein and DNA dynamics in murine C2C12 skeletal myotube cultures were determined. At 1000 U/ml, TNF induced 30% increases in protein and DNA content. The effects of TNF on protein accumulation were inhibited by aphidicolin, an inhibitor of DNA synthesis. IL-6 mimicked the effects of TNF on C2C12 cultures, inducing a 32% increase in protein accumulation and a 71% increase in the rate of protein synthesis. IL-6 also decreased expression of mRNA for several proteolytic system components, including ubiquitin 2.4 kb (51%) and 1.2 kb (63%), cathepsin B (39%) and m-calpain (47%), indicating that IL-6 acts on both protein synthesis and degradation. Incubation of murine C2C12 myotube cultures with TNF (1000 U/ml) in the presence of a polyclonal mouse anti-IL-6 antibody resulted in an abolishment of the effects of TNF on protein synthesis, but did not inhibit TNF-induced stimulation of DNA synthesis. These findings indicate that the effects of TNF on muscle protein synthesis are mediated by IL-6, but that TNF exerts IL-6 independent effects on proliferation of murine skeletal myoblasts. PMID- 11853881 TI - Interactions between regulatory and catalytic subunits of the Candida albicans cAMP-dependent protein kinase are modulated by autophosphorylation of the regulatory subunit. AB - The cAMP-dependent protein kinase (PKA) from Candida albicans is a tetramer composed of two catalytic subunits (C) and two type II regulatory subunits (R). To evaluate the role of a putative autophosphorylation site of the R subunit (Ser(180)) in the interaction with C, this site was mutated to an Ala residue. Recombinant wild-type and mutant forms of the R subunit were expressed in Escherichia coli and purified. The wild-type recombinant R subunit was fully phosphorylated by the purified C subunit, while the mutant form was not, confirming that Ser(180) is the target for the autophosphorylation reaction. Association and dissociation experiments conducted with both recombinant R subunits and purified C subunit showed that intramolecular phosphorylation of the R subunit led to a decreased affinity for C. This diminished affinity was reflected by an 8-fold increase in the concentration of R subunit needed to reach half-maximal inhibition of the kinase activity and in a 5-fold decrease in the cAMP concentration necessary to obtain half-maximal dissociation of the reconstituted holoenzyme. Dissociation of the mutant holoenzyme by cAMP was not affected by the presence of MgATP. Metabolic labeling of yeast cells with [(32)P]orthophosphate indicated that the R subunit exists as a serine phosphorylated protein. The possible involvement of R subunit autophosphorylation in modulating C. albicans PKA activity in vivo is discussed. PMID- 11853882 TI - Fructose inhibits apoptosis induced by reoxygenation in rat hepatocytes by decreasing reactive oxygen species via stabilization of the glutathione pool. AB - Oxidative stress induces apoptosis in liver parenchymal cells. The present study demonstrates that the substitution of fructose for glucose as sole carbon source in the incubation medium reduced apoptosis due to reoxygenation up to 50% in cultured rat hepatocytes. This anti-apoptotic action of fructose cannot be explained by the effects of this sugar on the intracellular ATP concentration and the ATP/ADP ratio. Rather, the suppression of apoptosis by fructose seems to be a consequence of remarkably higher intracellular levels of glutathione observed during reoxygenation in fructose-fed hepatocytes in contrast to glucose-fed ones. With fructose as substrate, the generation of excess reactive oxygen species (ROS) during the initial phase of reoxygenation was strongly reduced. With respect to ROS reduction and stabilization of the cellular glutathione pool fructose was found as efficient as a pretreatment of glucose fed cells with N acetyl-L-cysteine. The enhanced metabolization of ROS by the glutathione/glutathione peroxidase system in fructose-cultured hepatocytes under reoxygenation was expected to improve their mitochondrial status so that late events in the apoptotic pathway are suppressed. This could be confirmed by the reduced release of cytochrome c from mitochondria into the cytosol as well as by the observed decrease of caspase-3 activity during reoxygenation. PMID- 11853884 TI - Infection of cells with replication deficient adenovirus induces cell cycle alterations and leads to downregulation of E2F-1. AB - Gene products of recombinant replication-deficient adenovirus vectors of the first generation (Ad vector) can induce cell cycle dysregulation and apoptosis after infection in eukaryotic cells. The mechanisms underlying this complex process are largely unknown. Therefore, we investigated the regulation of the pRb/E2F-1 complex, which controls transition from G(0)/G(1) to S phase of the cell cycle. As Ad vector infection results in a decrease in the number of cells in G(0)/G(1) phase of the cell cycle, we observed a decline of the pRb protein level and, surprisingly, also a decrease of the E2F-1 protein and mRNA level in infected cell lines. Furthermore, in contrast to the reduction of cells in the G(0)/G(1) phase we observed increased protein levels of p53 and p21 proteins. However, as experiments in p53 deficient cell lines indicated, the decrease of pRb and E2F-1 is independent of p53 and p21 expression. Moreover, results obtained with Rb deficient cell lines indicated that the reduced E2F-1 expression is independent of pRb. These results suggest that Ad vector-induced cell cycle dysregulation is associated with a specific downregulation of E2F-1 independent of Rb and p53 genomic status of cells. PMID- 11853883 TI - Inhibition of somatostatin receptor 5-signaling by mammalian regulators of G protein signaling (RGS) in yeast. AB - Regulators of G-protein signaling (RGSs) are negative regulators of G-protein coupled receptor (GPCR)-mediated signaling that function to limit the lifetime of receptor-activated G(alpha)-proteins. Here we show that four mammalian RGSs differentially inhibit the activation of a FUS1--LacZ reporter gene by the STE2 encoded GPCR in yeast. In order to examine the role of the GPCR in modulating RGS function, we functionally expressed the human somatostatin receptor 5 (SST(5)) in yeast. In the absence of RGSs, FUS1--LacZ activation in response to somatostatin increased in a dose-dependent manner in cells expressing SST(5). In contrast to the results obtained with Ste2p, all RGSs completely inhibited SST(5)-mediated signaling even at concentrations of agonist as high as 10(minus sign5) M. The ability of RGSs to inhibit SST(5) signaling was further assessed in cells expressing modified Gpa1 proteins. Even though SST(5)-mediated FUS1--LacZ activation was 5-fold more efficient with a Gpa1p/G(i3alpha) chimera, response to somatostatin was completely abolished by all four RGSs. Furthermore, we demonstrate that RGS1, RGS2 and RGS5 have reduced ability to inhibit SST(5) mediated activation of the RGS-resistant Gpa1p(Gly302Ser) mutant suggesting that the ability to interact with the G(alpha)-protein is required for the inhibition of signaling. Taken together, our results indicate that RGSs serve as better GAPs for Gpa1p when activated by SST(5) than when this G-protein is activated by Ste2p. PMID- 11853885 TI - Expression and localization of AQP5 in the stomach and duodenum of the rat. AB - The expression, localization, and regulation of aquaporin 5 (AQP5), a member of the water channel family of proteins, was investigated in tissues of the rat gastrointestinal tract. Reverse transcriptase--polymerase chain reaction (RT- PCR) detected AQP5 mRNA in the lower stomach and duodenum. DNA sequencing confirmed that the cDNA fragment amplified had the complete sequence of the AQP5 cDNA fragment. Western blot analysis indicated the expression of a 27 kDa molecular mass AQP5 protein in the lower stomach and duodenum, which size was the same as that found for the protein in the submandibular gland and lungs. By immunohistochemistry using the IgG affinity-purified AQP5 antibody, the pyloric gland and Brunner's gland were primarily stained in the lower stomach and duodenum, respectively; a strong staining appeared in the apical and lateral membranes in both glands. These results indicate that AQP5 is present in the rat lower stomach and duodenum where it may be involved in a water transport mechanism. These results also support the idea that AQP5, and probably other aquaporins, are involved in water secretion in the stomach and duodenum although the volume of water transported via AQPs is unclear. PMID- 11853887 TI - CRE-mediated transcriptional activation is involved in cAMP protection of T-cell receptor-induced apoptosis but not in cAMP potentiation of glucocorticoid mediated programmed cell death. AB - Apoptosis of thymic cells induced by glucocorticoids (GC) and T-cell receptor (TCR) engagement are mutually antagonistic. We demonstrate that cAMP enhances GC and antagonizes TCR (anti-CD3) apoptosis on the same cell (DO-11.10 and 2B4.11 T cell hybridomas). We analyzed the activity of several transcription factors in this cAMP dual, stimulus-dependent, regulatory action. Anti-CD3 increases kB activity which is inhibited by CPTcAMP or dexamethasone (DEX), supporting the proapoptotic role of NFkB on TCR-induced apoptosis. Anti-CD3 not only increases kB- but diminishes GC response element (GRE)-activity induced by DEX, suggesting that TCR-mediated blockade of GC-induced apoptosis involves not only the proposed antiapoptotic action of NF-kB on GC, but also the inhibition of GRE-regulated proapoptotic genes. To test the involvement of CRE-driven transcription in the cAMP dual apoptotic regulation, cells were transfected with a CRE decoy DNA oligomer. Blockade of CRE transactivation with decoy targeting of CRE completely blocked the protection of TCR-induced apoptosis by cAMP, while it did not modify the enhancement by cAMP on GC-induced apoptosis. We show that CRE-binding factors have a definite role in T-cell apoptosis: they are involved in cAMP protection of TCR- but not in cAMP potentiation of GC-induced apoptosis. PMID- 11853886 TI - Intracellular pH regulation in U-2 OS human osteosarcoma cells transfected with P glycoprotein. AB - The molecular mechanisms responsible for intracellular pH regulation in the U2-OS osteosarcoma cell line were investigated by loading with 2',7'-bis(2 carboxyethyl)-5(6) carboxyfluorescein ester and manipulation of Cl(-) and Na(+) gradients, both in HEPES- and HCO(3)(-)/CO(2)-buffered media. Both acidification and alkalinisation were poorly sensitive to 4,4'-diisothiocyanate dihydrostilbene 2,2'-disulfonic acid, inhibitor of the anion exchanger, but sensitive to amiloride, inhibitor of the Na(+)/H(+) exchanger. In addition to the amiloride sensitive Na(+)/H(+) exchanger, another H(+) extruding mechanism was detected in U-2 OS cells, the Na(+)-dependent HCO(3)(-)/Cl(-) exchanger. No significant difference in resting pH(i) and in the rate of acidification or alkalinisation was observed in clones obtained from U-2 OS cells by transfection with the MDR1 gene and overexpressing P-glycoprotein. However, both V(max) and K' values for intracellular [H(+)] of the Na(+)/H(+) exchanger were significantly reduced in MDR1-transfected clones, in the absence and/or presence of drug selection, in comparison to vector-transfected or parental cell line. NHE1, NHE5 and at a lower extent NHE2 mRNA were detected in similar amount in all U2-OS clones. It is concluded that, although overexpression of P-glycoprotein did not impair pH(i) regulation in U-2 OS cells, the kinetic parameters of the Na(+)/H(+) exchanger were altered, suggesting a functional relationship between the two membrane proteins. PMID- 11853888 TI - Inhibition of the multidrug resistance P-glycoprotein activity by green tea polyphenols. AB - Many beneficial proprieties have been associated with polyphenols from green tea, such as chemopreventive, anticarcinogenic, antiatherogenic and antioxidant actions. In this study, we investigated the effects of green tea polyphenols (GTPs) and their principal catechins on the function of P-glycoprotein (P-gp), which is involved in the multidrug resistance phenotype of cancer cells. GTPs (30 microg/ml) inhibit the photolabeling of P-gp by 75% and increase the accumulation of rhodamine-123 (R-123) 3-fold in the multidrug-resistant cell line CH(R)C5, indicating that GTPs interact with P-gp and inhibit its transport activity. Moreover, the modulation of P-gp transport by GTPs was a reversible process. Among the catechins present in GTPs, EGCG, ECG and CG are responsible for inhibiting P-gp. In addition, EGCG potentiates the cytotoxicity of vinblastine (VBL) in CH(R)C5 cells. The inhibitory effect of EGCG on P-gp was also observed in human Caco-2 cells, which form an intestinal epithelial-like monolayer. Our results indicate that, in addition to their anti-cancer properties, GTPs and more particularly EGCG inhibit the binding and efflux of drugs by P-gp. Thus, GTPs or EGCG might be potential agents for modulating the bioavailability of P-gp substrates at the intestine and the multidrug resistance phenotype associated with expression of this transporter in cancer cells. PMID- 11853889 TI - Monomeric alcohol oxidase is preferentially digested by a novel protease from Candida boidinii. AB - A protease activity has been partially purified from peroxisomal matrix fractions of the methylotrophic yeast Candida boidinii. The enzyme migrates as a single peak on a sucrose velocity gradient with an apparent native molecular mass of approximately 80-90 kDa. Activity can be recovered from nonreducing sodium dodecyl sulfate gels as a approximately 20 kDa species, suggesting it is an oligomer. The protein exhibits chymotrypsin-like activity and cleaves the model compound suc-L-L-V-Y-AMC. Additionally, monomers of alcohol oxidase (AO), an abundant protein of C. boidinii peroxisomes, generated in vitro or in pulse radiolabeled cells, are preferentially sensitive to degradation by the protease. Sensitivity is lost over time in vivo as AO folds and matures into octamers, suggesting that the protease may be involved in these processes. PMID- 11853890 TI - Mechanism of tert-butylhydroperoxide induced apoptosis in rat hepatocytes: involvement of mitochondria and endoplasmic reticulum. AB - The purpose of the present work was to study the mechanisms involved in apoptosis induced by oxidative stress in rat hepatocytes. We focused on the apoptotic signaling molecules cytochrome c, Bcl-2 and Bax. Rat hepatocytes were exposed for 1 h to increasing concentrations of tert-butylhydroperoxide (t-BHP). Using lactate dehydrogenase (LDH) leakage as a biomarker for necrosis, and DNA fragmentation as a biomarker for apoptosis, we observed that a concentration of t BHP of 0.4-0.5 mM provides a transition point below which apoptosis is favored and beyond which necrosis is favored. Malondialdehyde and 8-oxo-guanine formation indicates that t-BHP induces oxidative stress and damage. However, at 0.4 mM t BHP, these oxidative molecular changes as well as LDH leakage no longer progress after the first hour of t-BHP exposure, suggesting the activation of some defense mechanisms. Western blot analysis of cytochrome c shows that its level increases in the cytosol while that of Bax decreases in this fraction as a result of t-BHP treatment. Moreover, there is a loss of Bcl-2 from mitochondria while, in contrast, Bax accumulates in this organelle following t-BHP treatment. However, cytochrome c appears to be relocalized to the endoplasmic reticulum as its presence in microsomes is greatly enhanced. We suggest that t-BHP triggers apoptosis through a step that involves cytochrome c release from mitochondria. This event is stimulated by Bcl-2 disappearance from mitochondria and Bax recruitment. Neutralization of excess cytosolic cytochrome c is achieved by its relocalization to the endoplasmic reticulum, hence triggering the down-regulation of apoptotic signals. PMID- 11853891 TI - Shock wave-mediated molecular delivery into cells. AB - A single shock wave generated by a shock tube is able to effectively deliver macromolecules such as fluorescein isothiocyanate-dextran into the cytoplasm of living cells without causing cytotoxicity. We report on the effect of varying the molecular weight of the dextran and the number of shock waves on the efficiency of delivery into a cancer cell line. The fraction of cells permeabilized and the total fluorescence delivered were measured by flow cytometry, and the cellular viability by a tetrazolium assay on adherent cells and these values were compared to cell permeabilization using digitonin. Shock waves can deliver molecules of up to 2000000 molecular weight into the cytoplasm of cells without toxicity and may have applications in gene therapy. PMID- 11853892 TI - A von Willebrand factor-derived heparin-binding peptide regulates cell--substrate adhesive strength and chemokinesis behavior. AB - The ability of a soluble heparin-binding oligopeptide sequence derived from the von Willebrand factor (vWF) to modulate the adhesion and chemokinetic migration behavior of arterial smooth muscle cells was assessed using a novel glass microsphere centrifugation assay and automated time-lapse fluorescence videomicroscopy, respectively. Treatment of cells grown on fibronectin-coated substrates with the heparin-binding peptide resulted in the disassembly of focal adhesions, as assessed by immunohistochemical staining. These observations were consistent with six-fold decrease in cell--substrate adhesive strength (P<0.001), a biphasic effect on migration speed (P<0.05), as well as a dose-dependent reduction in the percentage of motile cells and the cell dispersion coefficient (mu=S(2)T/2). The specificity of this response to the vWF-derived heparin-binding peptide was supported by the absence of an observed effect in the presence of either a scrambled peptide or a consensus heparin-binding peptide sequence of similar heparin affinity. These data support the notion that competitive interactions between cell surface heparan sulfates with heparin-binding peptide domains located in soluble peptide fragments may modulate chemokinetic cell migration behavior and other adhesion-related processes. PMID- 11853893 TI - Green tea polyphenol (-)-epigallocatechin 3-gallate inhibits MMP-2 secretion and MT1-MMP-driven migration in glioblastoma cells. AB - We have recently shown that green tea polyphenols, and especially (-) epigallocatechin 3-gallate (EGCg), acted as potent inhibitors of matrix metalloproteinase activities as well as of proMMP-2 activation (M. Demeule, M. Brossard, M. Page, D. Gingras, R. Beliveau, Biochim. Biophys. Acta 1478 (2000)). In the present work, we sought to examine the involvement of MT1-MMP in the EGCg induced inhibition of proMMP-2 activation. The incubation of U-87 glioblastoma cells in the presence of concanavalin A or cytochalasin D, two potent activators of MT1-MMP, resulted in proMMP-2 activation that was correlated with the cell surface proteolytic processing of MT1-MMP to its inactive 43 kDa form. Addition of EGCg strongly inhibited the MT1-MMP-dependent proMMP-2 activation. The inhibitory effect of EGCg on MT1-MMP was also demonstrated by the down-regulation of MT1-MMP transcript levels and by the inhibition of MT1-MMP-driven cell migration of transfected COS-7 cells. These observations suggest that this catechin may act at both the MT1-MMP gene and protein expression levels. In addition, treatment of cells with non-cytotoxic doses of EGCg significantly reduced the amount of secreted proMMP-2, and led to a concomitant increase in intracellular levels of that protein. This effect was similar to that observed using well-characterized secretion inhibitors such as brefeldin A and manumycin, suggesting that EGCg could also potentially act on intracellular secretory pathways. Taken together, these results indicate that EGCg targets multiple MMP mediated cellular events in cancer cells and provides a new mechanism for the anticancer properties of that molecule. PMID- 11853894 TI - Trial of the effects of an advice and relaxation tape given within the first 24 h of admission to hospital with acute myocardial infarction. AB - BACKGROUND: The study was performed to examine the effects of giving patients with acute myocardial infarction an advice and relaxation audio tape within 24 h of admission to hospital. METHODS: A prospective, two-group design was used with 243 subjects randomised to receive either the advice and relaxation tape or a music tape of their choice within 24 h of sustaining a myocardial infarction. Outcomes comprised anxiety, cardiac misconceptions, lifestyle change, attendance at a cardiac rehabilitation programme, and quality of life. RESULTS: Although the advice and relaxation tape reduced the number of cardiac misconceptions this did not lead to any improvements in outcome, whilst in hospital or at 6 months. Both tapes were equally appreciated by the patients, 98% of whom said that they would recommend them to other patients. CONCLUSIONS: An advice and relaxation tape reduces cardiac misconceptions but does not confer any other benefits over a music tape. PMID- 11853896 TI - Thoracoscopic monitoring for pericardial application of local drug or gene therapy. AB - Cardiovascular gene therapy is a promising new approach for a variety of diseases. As far as gene therapy aimed at the myocardium is concerned a new transcutaneous delivery method may be into the pericardial sac. OBJECTIVE: To evaluate the safety and applicability of the percutaneous pericardial delivery route for drug- or gene-therapy. METHODS: A percutaneous pericardial access device called a perducer was used to deliver either Indian ink or methylene blue to the pericardium of male yorkshire pigs under hemodynamic surveillance. Animals were sacrificed after either 3 h or 3 days. Post mortem distribution of the injectate was evaluated macroscopically and microscopically. RESULTS: With the perducer, the pericardial cavity was punctured in 10 pigs without hemodynamic complications. Although traces of dye could be seen in the pleural cavity in pigs sacrificed after 3 h, no evidence for dissemination was found in myocardial, lung, mediastinal lymph node and liver tissue in pigs sacrificed after 3 days. In two additional pigs the pericardium was punctured with the same perducer technique under simultaneous thoracoscopic monitoring. Visualization of the surface of the pericardium facilitated the procedure because pericardial fat could be avoided. CONCLUSION: Obtaining access to the pericardium with the perducer technique is safe and feasible. In addition thoracoscopic guidance may improve success rate and offers the possibility of on line surveillance. PMID- 11853897 TI - QT interval and final portion of T wave: measurements and dispersion in infants born at high altitude. AB - BACKGROUND: This research was conducted to determine whether the depolarization and repolarization cardiac process of children born at high altitude differs from that of children born at lower altitudes. METHODS: We studied three groups of 30 healthy newborns in the Venezuelan Andes. Group 3500 m consisted of infants born at 3500 m above sea level; Group 3000 m of infants born at 3000 m above sea level and Group 1600 m of infants born at 1600 m above sea level. Conventional 12-lead electrocardiograms were recorded in the 90 newborns. Corrected QT interval, duration from the peak to the end of the T wave, dispersion of QT interval and of the final portion of T wave and heart rate were computed in each subject. RESULTS: The findings of the present study show that infants born at high altitude (Groups 3500 and 3000 m) had a significantly longer corrected QT interval and a significantly shorter peak-to-end of T wave interval. Neither the heart rate nor the indexes of dispersion of the depolarization-repolarization process revealed abnormal values or significant differences among the groups. CONCLUSIONS: These results suggest that the depolarization and repolarization cardiac process of infants born at high altitude differs from that of children born at lower altitudes. PMID- 11853898 TI - Long-term results after acute percutaneous transluminal coronary angioplasty in acute myocardial infarction and cardiogenic shock. AB - The aim of this study was to determine the long-term outcome in unselected, consecutive patients after acute percutaneous transluminal angioplasty (PTCA) for acute myocardial infarction (AMI) complicated by cardiogenic shock. This involved a follow-up study from a prospectively conducted patient registry in a tertiary referral center. A total of 59 patients (10 female/49 male; median age 62 years (32-91)) with percutaneous transluminal cardiac interventions in primary cardiogenic shock were identified between January 1995 and January 2000. Twenty two patients (37%) had been resuscitated successfully before intervention. The in hospital mortality of shock patients was 36% (n=21, median age 68 (47-84)). The median follow-up of survivors was 18.1 (7-57.3) months, during which three further patients died (8%; two because of sudden cardiac deaths, one because of acute reinfarction). Achievement of thrombolysis in myocardial infarction (TIMI) flow III after acute PTCA (84% in survivors vs. 38% in non-survivors; P<0.001) and the absence of the left main coronary artery (3% survivors vs. 29% non survivors; P=0.003) as culprit lesion in patients with cardiogenic shock was strongly associated with an improved survival rate. A second cardiac intervention was performed in seven patients (18%). Overall functional capacity of shock survivors was good. At final follow-up, 80% of the survivors were completely asymptomatic. One patient had angina pectoris NYHA II, five patients dyspnoea NYHA class II. Exercise stress-test was performed in 24 of the 38 surviving patients, median exercise capacity was 100% (range 55-113%) of the age adjusted predicted value. In unselected patients with cardiogenic shock due to AMI, treatment with acute PTCA resulted in an in-hospital mortality of 36%, low late mortality and good functional capacity in long-term survivors. TIMI flow grade III after acute PTCA in patients with acute myocardial infarction complicated by cardiogenic shock was strongly associated with an improved survival rate whereas the left main coronary artery as culprit lesion was associated with worse outcome. PMID- 11853899 TI - Heme oxygenase-1 pathway is involved in delayed protection induced by heat stress against cardiac ischemia-reperfusion injury. AB - Previous studies have shown that heme oxygenase-1 (HO-1), a heat stress protein (HSP32), has a beneficial effect on the ischemic myocardium. The purpose of the present study was to explore whether HO-1 is involved in delayed cardioprotection provided by heat stress in vivo. Sprague--Dawley rats were pretreated with whole body hyperthermia (rectal 42 degrees C) for 15 min followed by ischemia reperfusion 24 h later. Ischemia-reperfusion injury was induced by 45 min of coronary artery occlusion followed by a 3-h reperfusion. Myocardial injury degree was evaluated by measurement of infarct size and serum creatine kinase (CK) activity. The expression of HO-1 mRNA and protein in myocardial tissues were measured. Pretreatment with hyperthemia significantly reduced infarct size and CK release during reperfusion, which was completely blocked by pretreatment with ZnPP-9, an inhibitor of HO and methylene blue, an inhibitor of soluble guanylate cyclase. Heat stress also significantly increased the expression of HO-1 mRNA and protein, and the effect was not affected by pretreatment with methylene blue. The present results suggest that the HO-1 pathway is involved in the mediation of delayed cardioprotection by heat stress in rats. PMID- 11853900 TI - The association of Mediterranean diet with lower risk of acute coronary syndromes in hypertensive subjects. AB - BACKGROUND: The elevation of blood pressure levels has been recognised as a determinant of the risk for several common cardiovascular diseases. In this work we examined the effect of Mediterranean type of diet on coronary risk in subjects with hypertension. METHODS: CARDIO2000 consisted of 848 randomly selected hospitalised patients (695 males, 58 +/- 10 years old; 153 females, 65 +/- 9 years old) for first event of coronary heart disease (CHD) and 1078 paired, by sex-age, hospitalised controls without CHD. The adoption of the Mediterranean diet was assessed through a validated questionnaire developed by the National School of Public Health. RESULTS: 418 (49%) of the patients and 303 (28%) of the controls were hypertensive. Of them 21 (5%) patients and 36 (12%) controls were unaware of their condition, 94 (22%) and 34 (11%) were untreated, 148 (35%) and 111 (36%) were uncontrolled and 155 (38%) and 122 (41%) were controlled (P<0.01). One hundred and sixty-two (19%) of the patients and 265 (25%) of the controls (P<0.01) adopted the Mediterranean type of diet. Our results suggest that the adoption of Mediterranean diet reduces the risk of developing acute coronary syndromes by 17% (odds ratio=0.83, 95% CI 0.73--0.88, P<0.01) in controlled hypertensive subjects, by 8% (odds ratio=0.92, 95% CI 0.87-0.95, P<0.05) in unaware, by 7% (odds ratio=0.93, 95% CI 0.88-0.95, P<0.05) in acknowledged but uncontrolled and by 20% (odds ratio=0.80, 95% CI 0.71-0.89, P<0.01) in normotensive subjects. CONCLUSION: According to our findings the adoption of the Mediterranean diet is associated with the reduction of coronary risk in hypertensive subjects. PMID- 11853901 TI - Current evidence supporting the role of diuretics in heart failure: a meta analysis of randomised controlled trials. AB - OBJECTIVE: To summarise the current evidence from randomised controlled trials for diuretics in patients with congestive heart failure (CHF). DATA SOURCES: English-language randomised controlled trials and review papers referenced in Medline, Embase between 1966 and 1999. General literature review of pertinent journals was carried out and reference lists of papers were inspected. REVIEW METHOD: STUDY DESIGN: Meta-analysis of randomised controlled trials of diuretic therapy in patients with CHF. STUDY SELECTION: Studies were included if they were randomised comparisons of loop or thiazide diuretics and control, or one diuretic and another active agent (e.g. ACE inhibitors, ibopamine and digoxin). DATA ABSTRACTION: Using a standardised protocol, two reviewers independently abstracted the data and assessed the methodological quality of each paper. DATA SYNTHESIS: The odds ratio (OR) of treated group compared with control was estimated for each end-point outcome and plotted against each other using the fixed-effects model. THE MAIN OUTCOME MEASURES: The primary outcomes of our analysis were effects of diuretics on mortality and morbidity. RESULTS: Eighteen trials met our criteria and were eligible for analysis, involving 928 patients. Eight trials were placebo-controlled. We analysed the data for mortality and for worsening heart failure. A further ten trials compared diuretics against other agents such as ACE inhibitors, ibopamine, and digoxin. Mortality data were available in three of the placebo-controlled trials (n=221); the mortality rate was lower for patients treated with diuretics than for control [the odds ratio for death, 0.25; 95% confidence intervals (CI), 0.07-0.84; P=0.03]. Admissions for worsening heart failure in the four small trials (n=448) showed an odds ratio of 0.31 (95% CI 0.15-0.62; P=0.001). In six studies of diuretics compared to active control, diuretics significantly improved exercise capacity in patients with CHF [OR: 0.37; CI: 0.10-0.64, P=0.007]. CONCLUSION: Compared to active control, diuretics appear to reduce the risk of worsening disease and improve exercise capacity. The available data from small studies show that in CHF conventional diuretics reduce the risk of death and worsening heart failure compared to placebo. PMID- 11853902 TI - Electrocardiographic associations of right precordial Q waves help to distinguish anterior myocardial infarction from aortic stenosis. AB - Right precordial Q waves are ECG evidence of anterior myocardial infarction and can be present in patients with pathological left ventricular hypertrophy particularly caused by aortic stenosis. The aim of this study was to investigate the ECG features associated with Q waves in aortic stenosis and those in anterior myocardial infarction. We studied 16 patients with anterior myocardial infarction and 19 patients with aortic stenosis by means of ECG, echocardiography and clinical history. On the ECG, heart rate (70 +/- 20 beats/min vs. 83 +/- 20) and QT interval (380 +/- 65 ms vs. 390 +/- 50) did not differ between the two conditions. PR interval (160 +/- 15 ms vs. 185 +/- 30, P<0.05) and QRS duration (80 +/- 7.0 ms vs. 95 +/- 15, P<0.01) were both longer in patients with aortic stenosis than in those with myocardial infarction. The Q wave voltage in V1 (1.0 +/- 0.55 mV vs. 1.5 +/- 0.60) or V2 (1.3 +/- 0.5 mV vs. 1.8 +/- 0.85) and R wave voltage in V5 (0.7 +/- 0.7 mV vs. 2.1 +/- 0.9) or V6 (0.7 +/- 0.4 mV vs. 1.5 +/- 0.7, all P<0.01) were significantly less in patients with anterior myocardial infarction than in those with aortic stenosis. Q wave voltage over 1.3 mV in V1 or R wave voltage over 1.5 mV in V5 can differentiate aortic stenosis from anterior myocardial infarction with a sensitivity of 79% for each and specificities of 81 and 93.8%, respectively. Though the frontal QRS axis was similar in the two groups (28 +/- 45 degrees vs. 14 +/- 35, P>0.05), the horizontal QRS axis pointed laterally (-30 +/- 20 degrees) in aortic stenosis and posteriorly (-60 +/- 20 degrees, P<0.01) in anterior myocardial infarction. A horizontal QRS axis between zero and -45 degrees detected the presence of aortic stenosis with a sensitivity of 94.7% and a specificity of 81.3%. On echocardiography, left ventricular hypertrophy was found in most patients (94.7%) with aortic stenosis but not in those (0%) with anterior myocardial infarction. Left ventricular end diastolic dimensions (5.1 +/- 0.7 cm vs. 5.1 +/- 0.9, P>0.05) were similar in the two groups but the end systolic dimension was increased in patients with aortic stenosis (4.0 +/- 0.9 cm vs. 3.4 +/- 0.6, P<0.05). The systolic left ventricular function (shortening fraction: 23 +/- 8.0% vs. 34 +/- 7.0; Vcf: 0.8 +/- 0.26 circ/s vs. 1.3 +/- 0.26, both P<0.01) was significantly impaired in patients with aortic stenosis compared to those with myocardial infarction. In conclusion, in the presence of right precordial Q waves, the simple 12-lead ECG can provide important information on distinguishing anterior myocardial infarction from aortic stenosis. In particular, the QRS voltage in the chest leads and horizontal QRS axis can differentiate anterior myocardial infarction from aortic stenosis with high sensitivity and specificity. PMID- 11853903 TI - Atrial septal pacing in the prevention of paroxysmal atrial fibrillation refractory to antiarrhythmic drugs. AB - BACKGROUND: Early reports have shown that pacing the atria at a site or sites other than the right atrial appendage may prevent atrial fibrillation. Our centre has shown that pacing the atrial septum reduces the duration of atrial activation which is an important determinant of predisposition to paroxysmal atrial fibrillation. Ablation of the atrioventricular (AV) node together with implantation of a pacemaker can control symptoms due to paroxysmal atrial fibrillation in patients in whom antiarrhythmic drugs have failed. The aim of this study was to investigate the effect of atrial septal pacing on patients who were candidates for AV node ablation. METHODS: Atrial septal pacemakers were implanted in 28 patients with symptomatic, paroxysmal atrial fibrillation that had been unresponsive to two or more antiarrhythmic drugs. Pacing was not indicated for any reason other than the anticipated need to proceed to AV node ablation. Change in symptoms was assessed by quality of life questionnaires and recurrence of atrial fibrillation was measured objectively by pacemaker interrogation and ambulatory electrocardiographic monitoring. RESULTS: Atrial septal pacing in combination with an antiarrhythmic agent resulted in a substantial subjective improvement in 19 patients (68%). Objective data confirmed similar findings; atrial fibrillation was completely or markedly reduced in 17 patients (60%). Six patients experienced a modest improvement in symptoms; in only four patients was it necessary to proceed to AV node ablation. CONCLUSIONS: Atrial septal pacing together with continuance of previously ineffective antiarrhythmic therapy may prevent or markedly reduce the frequency of paroxysmal atrial fibrillation and obviate the need to ablate the AV node. PMID- 11853904 TI - A case of severe respiratory depression due to cibenzoline overdosage induced by a transient renal dysfunction. AB - A 69-year-old man was transferred to our hospital because of severe general fatigue and progressive systemic muscle weakness. He had taken 300 mg/day of cibenzoline for his sustained ventricular tachycardia (VT) for years. At the end of June 2001 he began to feel general fatigue, which slowly progressed to systemic muscle weakness and walking disturbance. On 2nd July 2001, he finally could not stand up by himself. He also felt dyspnea. He was transferred to our emergency room, where he developed severe respiratory depression. Acute myocardial infarction was ruled out based on his ECG and blood chemistry data. Serum BUN and creatinine were elevated to 32 and 2.2 mg/dl, respectively, which returned to normal range 2 weeks later. What we did in our ICU were basically a replacement of cibenzoline with mexiletine and mechanical support of ventilation. As his renal function gradually improved, his spontaneous respiration and muscle power were slowly restored. He was discharged on foot after 1 month of hospitalization. His blood cibenzoline content taken 2 days after the cessation of cibenzoline was 959.6 mg/ml that was abnormally elevated. Considering metabolism and excretion for the 2 days between the cibenzoline cessation and the blood sample drawing, his cibenzoline level on the day of admission must have been extraordinary high. We should be aware of the possibility of abrupt overdosage of cibenzoline even in patients with normal kidney function in the event of a transient or an acute renal dysfunction. PMID- 11853906 TI - Multiple pseudoaneurysms of the femoral artery. PMID- 11853905 TI - Apical hypertrophic cardiomyopathy, intraventricular pressure gradients and ST segment elevation. PMID- 11853907 TI - Myocardial ischaemia with a normal coronary angiogram due to the primary antiphospholipid syndrome. AB - In this case report, we describe a 33-year-old woman with a history of two unprovoked thrombo-embolic events presenting with acute myocardial ischaemia. She had a normal coronary angiogram (CAG). The diagnosis primary antiphospholipid syndrome (APS), an acquired hypercoagulability disorder, was established by the presence of antibodies directed against phospholipids. The primary APS should be considered as a cause of myocardial ischaemia in patients with a normal CAG and a history of unprovoked thrombo-embolic events. PMID- 11853908 TI - Calcified left ventricular thrombus in a patient after anterior myocardial infarction. PMID- 11853909 TI - Nontyphoidal salmonella pericarditis: a case of Salmonella typhimurium phage type 2 pericarditis. PMID- 11853910 TI - Unusual complication of left heart catheterisation. PMID- 11853911 TI - Thrombolysis for acute massive pulmonary embolism during pregnancy. PMID- 11853914 TI - A cationic sub-micron emulsion (MF59/DOTAP) is an effective delivery system for DNA vaccines. AB - A novel cationic emulsion was developed to adsorb plasmid DNA and improve intracellular delivery of plasmid DNA upon immunization. The emulsion developed, had a highly uniform particle and charge distribution. Based on observations with cationic microparticles, the cationic emulsion was evaluated in vivo in mice and rabbits with a model HIV-1 pCMVp55 gag DNA. In both these species, the cationic emulsion engendered higher antibody responses than those obtained with naked DNA. The cationic emulsion also maintained the cellular responses seen with naked DNA at the same doses. PMID- 11853915 TI - Formulation aspects in the development of osmotically controlled oral drug delivery systems. AB - Osmotically controlled oral drug delivery systems utilize osmotic pressure for controlled delivery of active agent(s). Drug delivery from these systems, to a large extent, is independent of the physiological factors of the gastrointestinal tract and these systems can be utilized for systemic as well as targeted delivery of drugs. The release of drug(s) from osmotic systems is governed by various formulation factors such as solubility and osmotic pressure of the core component(s), size of the delivery orifice, and nature of the rate-controlling membrane. By optimizing formulation and processing factors, it is possible to develop osmotic systems to deliver drugs of diverse nature at a pre-programmed rate. In the present review, different types of oral osmotic systems, various aspects governing drug release from these systems, and critical formulation factors are discussed. PMID- 11853916 TI - Targeting to macrophages: role of physicochemical properties of particulate carriers--liposomes and microspheres--on the phagocytosis by macrophages. AB - Heterogeneous functions of macrophages in human immune systems have renewed interest in targeting of drugs to these cells. Various carrier systems have emerged to deliver drugs to macrophages, albeit the efficacy, reliability and selectivity of these carriers are still in question. To date, the most extensively studied carriers are liposomes and microspheres. Various physicochemical properties of these carriers can alter their efficacy and specificity to a great extent. These properties include hydrophilicity, surface charge, composition, concentration, and presence of various ligands. In this review, a comprehensive evaluation of the literature has been carried out in order to show the role of these parameters in the design of carriers for targeting of drugs to macrophages. PMID- 11853917 TI - Anticancer drug delivery systems: N4-acyl poly(ethyleneglycol) prodrugs of ara-C. I. Efficacy in solid tumors. AB - A systematic study of N(4) amino PEG-prodrugs of ara-C (1) was conducted and provided a series of disubstituted amides, as well as a carbamate derivative. These conjugates showed hydrolysis half lives in rat plasma from about 1 h to 3 days, but were stable for >24 h in phosphate buffer, pH 7.4. In an LX-1 solid lung tumor model some of the PEG prodrugs exhibited superior activity to ara-C when compared on a molar basis. One problematic issue that was identified in this investigation was the need to increase the loading of ara-C onto PEG in order to avoid highly viscous solutions. PMID- 11853918 TI - Anticancer drug delivery systems: multi-loaded N4-acyl poly(ethylene glycol) prodrugs of ara-C. II. Efficacy in ascites and solid tumors. AB - The synthesis of branched PEG (40,000) acids has been achieved using aspartic acid (Asp) and AspAsp dendrons. Complete conjugation of these dendritic acids with cytosine arabinoside (ara-C) was achieved by the use of spacers that allowed a greater separation of the branches to accommodate several large ara-C molecules in proximity to each other. The tetrameric and octameric PEG-ara-C amide prodrugs were much more effective in the treatment of solid and ascites tumors compared to the native drug. The greater loading of the PEG backbone appears to have achieved a minimum threshold concentration for the therapeutic delivery of ara-C. PMID- 11853919 TI - A floating-type oral dosage form for piroxicam based on hollow polycarbonate microspheres: in vitro and in vivo evaluation in rabbits. AB - A floating type dosage form (FDF) of piroxicam in hollow polycarbonate (PC) microspheres capable of floating on simulated gastric and intestinal fluids was prepared by a solvent evaporation technique. Incorporation efficiencies of over 95% were achieved for the encapsulation. In vitro release of piroxicam from PC microspheres into simulated gastric fluid at 37 degree C showed no significant burst effect. The amount released increased with time for about 8 h after which very little was found to be released up to 24 h. In intestinal fluid, the release was faster and continuous and at high drug payloads, the cumulative release reached above 90% in about 8 h. In vivo evaluation of different dosage forms of piroxicam such as free drug, drug-encapsulated microspheres and microspheres along with a loading dose of free drug in rabbits showed multiple peaking in the plasma concentration-time curve suggesting enterohepatic recirculation of the drug. Pharmacokinetic analysis showed that the bioavailability from PC microspheres alone was about 1.4 times that of the free drug and it was about 4.8 times for the dosage form consisting of the microspheres plus the loading dose. The elimination half life was increased by about three times for the microsphere preparation alone and nearly about six times for the dosage form comprising of microspheres and a loading dose in comparison to the free drug. Data obtained in this study demonstrated that FDF of piroxicam in PC microspheres was capable of sustained delivery of the drug for longer periods with increased bioavailability. PMID- 11853920 TI - A novel sustained-release formulation of insulin with dramatic reduction in initial rapid release. AB - To ensure a strictly controlled release of insulin, a preparation method for insulin-loaded microcapsules was designed. Microcapsules were prepared with an injectable, biodegradable polymer composed of co-poly(D,L-lactic/glycolic) acids (PLGA) (mean molecular weight 6600, LA/GA ratio 50:50). Morphological examination using scanning electron microphotography demonstrated spherical particles with a main diameter of 15-30 microm. When 3% insulin-loaded PLGA microcapsules were administered subcutaneously as a single dose (250 U/kg) to streptozotocin-induced hyperglycemic rats, plasma insulin levels increased and were sustained at levels showing hypoglycemic effects. When glycerin, ethanol, or distilled water was used throughout the preparation procedure, the resultant microcapsules dramatically reduced the initial burst. The formulation in which glycerin was added to an oil phase containing PLGA, insulin, and ZnO increased plasma insulin levels to 86.7, 108.4, and 84.9 microU/ml at 1, 2, and 6 h, respectively. The levels remained at 36.2-140.7 microU/ml from day 1 to day 9. The AUC(0-24 h)/AUC(0-336 h) ratio was calculated to be 9.7%. The formulation prepared without additives gave such a rapid insulin release that animals receiving it became transiently hypoglycemic. PMID- 11853921 TI - Transdermal drug delivery of imipramine hydrochloride. I. Effect of terpenes. AB - The objective of this investigation was to study the effect of different terpenes on IMH permeation in EtOH:W (2:1) system. Permeation studies of IMH were carried out with unjacketed Franz diffusion cells through rat skin. The flux of IMH with terpenes was found to be significantly higher than that in control (EtOH:W, 2:1) (P<0.05). Amongst all studied terpenes, menthol, terpineol, cineole and menthone were found to be effective permeation enhancers for IMH. It was found that the contribution of diffusivity in enhanced permeation of IMH was much higher in comparison to partitioning of IMH in skin with terpene treatment. Results of this study were explained with the help of H-bond breaking potential and self association of terpenes. In order to elucidate the effect of terpenes on stratum corneum barrier FT-IR was used. PMID- 11853922 TI - Colon-specific delivery of prednisolone-appended alpha-cyclodextrin conjugate: alleviation of systemic side effect after oral administration. AB - Prednisolone (PD), a typical glucocorticoid, has been widely used for the treatment of inflammatory bowel disease (IBD). However, when PD is administered orally, a large amount of the drug is absorbed from the upper gastrointestinal (GI) tract and causes systemic side effects. In this study, the anti-inflammatory effect and systemic side effect of the PD succinate/alpha-cyclodextrin (PDsuc/alpha-CyD) ester conjugate after oral administration were studied using IBD model rats. The anti-inflammatory effect of the PDsuc/alpha-CyD conjugate was comparable to those of PD alone. On the other hand, the systemic side effect of the PDsuc/alpha-CyD conjugate was much lower than that of PD alone when administered orally. The lower side effect of the conjugate was attributable to passage of the conjugate through the stomach and small intestine without significant degradation or absorption, followed by the degradation of the conjugate site-specifically in the large intestine. The oral administration of PD alone gave higher plasma concentrations of PD, giving the significant systemic side effect. The results suggested that the PDsuc/alpha-CyD conjugate is useful as a delayed-release type prodrug of PD for colon-specific delivery, owing to alleviation of the systemic side effect, while maintaining the therapeutic effect. PMID- 11853923 TI - Percutaneous penetration and skin metabolism of ethylsalicylate-containing agent, TU-2100: in-vitro and in-vivo evaluation in guinea pigs. AB - The aim of this study was to investigate the percutaneous penetration and dermal metabolism of a new potential anti-acne prodrug--TU-2100 [bis(o-carboxyphenyl ethyl ester)nonanedioate] in guinea pigs. The fluxes of this agent through excised skin after applications of TU-2100 gels at 3 and 10% concentrations were similar. However, after 24 h from the time of drug application, the total amounts of permeated TU-2100 into the skin compartment and through the skin into the receiver were 271.7 (+/-30.7 S.E.) microg/cm(2) from the 3% gel and 779.4.0 (+/ 98.5 S.E.) microg/cm(2) from the 10% gel, demonstrating a relatively high skin accumulation. Higher degradation of TU-2100 to ethylsalicylate occurred after application of drug at 10% concentration than after the application of 3% gel. In contrast, the fraction of permeated drug metabolized was twofold higher after the 3% gel application than after the 10% gel (F(m)=20 vs. 10.5 mole %). Since F(m) is reversibly related to the total permeating drug, the obtained values actually reflect the significant difference in TU-2100 permeation from the 3% (271.7 microg) and the 10% (779.4 microg) gels. An in vivo--in vitro comparison revealed similar drug accumulations in the skin after application of both 3 and 10% gels, however, skin metabolism was found to be significantly higher in vivo than in vitro. PMID- 11853924 TI - PLGA-mPEG nanoparticles of cisplatin: in vitro nanoparticle degradation, in vitro drug release and in vivo drug residence in blood properties. AB - The in vitro nanoparticle degradation, in vitro drug release and in vivo drug residence in blood properties of PLGA-mPEG nanoparticles of cisplatin were investigated. The nanoparticles were prepared by a double emulsion method and characterized with regard to their morphology, size, zeta potential and drug loading. The rate of in vitro degradation of the PLGA-mPEG nanoparticles in PBS (pH 7.4) depended on their composition, increasing when the mPEG content (mPEG:PLGA ratio) of the nanoparticles increased. Sustained cisplatin release over several hours from the PLGA-mPEG nanoparticles in vitro (PBS) was observed. The composition of the nanoparticles affected drug release: the rate of release increased when the mPEG content of the nanoparticles increased. Within the range of drug loadings investigated, the drug loading of the nanoparticles did not have any significant effect on drug release. The loading efficiency was low and needs improvement in order to obtain PLGA-mPEG nanoparticles with a satisfactory cisplatin content for therapeutic application. The i.v. administration of PLGA mPEG nanoparticles of cisplatin in BALB/c mice resulted in prolonged cisplatin residence in systemic blood circulation. The results appear to justify further investigation of the suitability of the PLGA-mPEG nanoparticles for the controlled i.v. delivery and/or targeting of cisplatin. PMID- 11853925 TI - Assessment of protein release kinetics, stability and protein polymer interaction of lysozyme encapsulated poly(D,L-lactide-co-glycolide) microspheres. AB - Using lysozyme as a model protein, this study investigated protein stability, protein--polymer interaction in different release media and their influence on protein release profile and in vitro--in vivo correlation. Lysozyme was microencapsulated into PLGA 50:50 by a double emulsion--solvent extraction/evaporation method. Protein stability, protein--PLGA adsorption and protein in vitro release were studied in various test media. Differential scanning calorimetry analysis showed lysozyme to be most conformationally stable in pH 4.0 acetate buffer with highest T(m) at 77.2 degree C and DeltaH(cal) 83.1 kcal/mol. Lysozyme exhibited good stability in pH 2.5 glycine buffer with T(m) at 63.8 degree C and DeltaH(cal) 69.9 kcal/mol. In pH 7.4 phosphate-buffered saline (PBS), lysozyme showed a trend toward aggregation when the temperature was elevated. When PLGA polymer was incubated with lysozyme in the various buffers, adsorption was found to occur in PBS only. The adsorption severely limited the amount of lysozyme available for release from microspheres, resulting in slow and incomplete release in PBS. In contrast, the release of the microspheres in acetate and glycine buffers was complete within 40 and 70 days, respectively. Radiolabeled lysozyme blood levels in rats from the microspheres correlated qualitatively well with in vitro release in glycine buffer as a release medium. This study suggests that protein stability and adsorption are critical factors controlling protein release kinetics and in vitro--in vivo correlation of PLGA microspheres. PMID- 11853926 TI - The enhancing effect of nasal absorption of FITC-dextran 4,400 by beta-sitosterol beta-D-glucoside in rabbits. AB - The effect and mechanism of action of beta-sitosterol beta-D-glucoside (Sit-G) on the in vitro and in vivo nasal absorption of FITC-dextran (molecular weight, 4400; FD-4) in rabbits were studied in comparison with beta-sitosterol (Sit). The FD-4 permeation in the powder dosage form was increased by Sit-G and Sit and related to the uptake of Sit-G and Sit with no changes in the amount of cholesterol in the excised nasal mucosa. The application of Sit and Sit-G increased FD-4 permeation with and without a decrease in transepithelial resistance (TEER), respectively. These results suggested that the mechanism of the enhancement by Sit-G was different from those of Sit and sodium caprate; Sit G may exert its effects mainly via the transcellular pathway due to perturbation of the mucosal membrane. PMID- 11853927 TI - Modulation of active pharmaceutical material release from a novel 'tablet in capsule system' containing an effervescent blend. AB - The objective of the present study was to obtain programmed drug delivery from hard gelatin capsules containing a hydrophilic plug (HPMC or guar gum). The significance of factors such as type of plug (powder or tablet), plug thickness and the formulation of fill material on the release pattern of diltiazem HCl, a model drug, was investigated. The body portion of the hard gelatin capsules was cross-linked by the combined effect of formaldehyde and heat treatment. A linear relationship was observed between weight of HPMC K15M and log % drug released at 4 h from the capsules containing the plug in powder form. In order to accelerate the drug release after a lag time of 4 h, addition of an effervescent blend, NaHCO(3) and citric acid, in the capsules was found to be essential. The plugs of HPMC in tablet form, with or without a water soluble adjuvant (NaCl or lactose) were used for obtaining immediate drug release after the lag time. Sodium chloride did not cause significant influence on drug release whereas lactose favourably affected the drug release. The capsules containing HPMC K15M tablet plug (200 mg) and 35 mg effervescent blend in body portion of the capsule met the selection criteria of less than 10% drug release in 4 h and immediate drug release thereafter. It is further shown that the drug release was also dependant on the type of swellable hydrophilic agent (HPMC or guar gum) and molecular weight of HPMC (K15M or 20 cPs). The results reveal that programmed drug delivery can be obtained from hard gelatin capsules by systemic formulation approach. PMID- 11853928 TI - The effect of fatty acid substitution on the in vitro release of amphotericin B from micelles composed of poly(ethylene oxide)-block-poly(N-hexyl stearate-L aspartamide). AB - The effect of fatty acid substitution on the in vitro release of amphotericin B (AmB) from micelles composed of poly(ethylene oxide)-block-poly[N-(6-hexyl stearate)-L-aspartamide] (PEO-b-PHSA) was investigated. PEO-b-PHSA at 11, 50 and 70% of stearic acid substitution self assembled into micelles that effectively encapsulate AmB by solvent evaporation and dialysis methods. The sustained release of AmB from PEO-b-PHSA micelles was evidenced, by measuring the transfer of the drug to lipid vesicles [dipalmitoyl phosphatidylcholine:cholesterol:dimyristoyl phosphatidyglycerol (3:1:0.25)]. The release of AmB for PEO-b-PHSA micelles was markedly influenced by the degree of fatty acid substitution--as it increased, the release of AmB slowed. Accordingly, drug release was found to correlate with haemolysis induced by AmB encapsulated in PEO-b-PHSA micelles. At 11% stearic acid substitution, encapsulation of AmB had little effect on the drug's ability to induce untoward haemolysis. In contrast, AmB stably encapsulated in PEO-b-PHSA micelles at 50 and 70% caused no hemolysis up to 20 microg/ml. Lastly, PEO-b-PHSA micelles at 50 and 70% were able to elute entirely as micelles during size-exclusion chromatography, indicating their stability toward dissociation after dilution. The results point to a nanoscopic drug depot that may release AmB at controlled rates. PMID- 11853929 TI - Ex vivo bioadhesion and in vivo testosterone bioavailability study of different bioadhesive formulations based on starch-g-poly(acrylic acid) copolymers and starch/poly(acrylic acid) mixtures. AB - Starch-g-poly(acrylic acid) copolymers or grafted starches synthesized by 60Co irradiation or chemical modification and co-freeze-dried starch/poly(acrylic acid) mixtures were evaluated on their ex vivo bioadhesion capacity. The buccal absorption of testosterone from a bioadhesive tablet formulated with the grafted starches or starch/poly(acrylic acid) mixtures was investigated. The results were compared to a reference formulation (physical mixture of 5% Carbopol 974P and 95% Drum Dried Waxy Maize). Rice starch-based irradiated grafted starches showed the best bioadhesion results. Partial neutralization of the acrylic acid with Ca(2+) ions resulted in significantly higher bioadhesion values compared to the reference. Ca(2+) and Mg(2+) partially neutralized maltodextrin-based irradiated grafted starches showed significantly higher bioadhesion values compared to the reference formulation. The chemically modified grafted starches showed significantly higher adhesion force values than for the reference tablet. None of the co-freeze-dried starch/poly(acrylic acid) mixtures showed significantly higher bioadhesion results than the reference (Bonferroni test, P<0.05). A chemically modified grafted starch could sustain the 3 ng/ml plasma testosterone target concentration during +/- 8 h (T(>3 ng/ml)). By lyophilization of a partially neutralized irradiated grafted starch, the in vivo adhesion time (22.0 +/- 7.2 h) and the T(>3 ng/ml) (13.5 +/- 1.3 h) could be increased. The absolute bioavailability of the lyophilized formulation approached the reference formulation. Some of the grafted starches showed to be promising buccal bioadhesive drug carriers for systemic delivery. PMID- 11853930 TI - Lectin-mediated drug delivery: influence of mucin on cytoadhesion of plant lectins in vitro. AB - As the mucous layer represents the first barrier to peroral lectin-mediated drug delivery, the influence of mucin on the cytoadhesive properties of lectins was studied in vitro by establishing a rapid and simple microplate format assay using pig gastric mucin (PGM) for coating the wells. The lectin-binding capacity of mucin followed the order WGA>>UEA-I>>LCA=STL>PNA>DBA. The PGM-binding of wheat germ agglutinin (WGA) was strongly dependent on pH being highest at pH 5.0. In comparison, PGM-binding of WGA was about 15% at gastric pH and 60-70% at intestinal pH. This points to unimpeded gastric transit of WGA-grafted formulations and favorable conditions within the intestine for binding to mucus coated enterocytes. Moreover the WGA-PGM interaction was concentration-dependent, specific and fully reversible. According to a competitive assay in the presence of Caco-2 monolayers, the PGM-binding of WGA was saturated and influenced by the lectin-concentration yielding 28% Caco-2 bound WGA (125 ng WGA/0.29 cm(2) monolayer) and 68% Caco-2 bound WGA (4 microg WGA/0.29 cm(2) monolayer), respectively. Following on from these results, lectins are expected to suffer at least partially from premature inactivation by shed off mucus like bioadhesives of the first generation, however initial but reversible mucus-binding of lectins offers partititioning to the cell membrane followed by uptake into the enterocyte. PMID- 11853931 TI - Double emulsions: how does release occur? AB - Water-in-oil-in-water double emulsions (W/O/W) consist of dispersed oil globules containing smaller aqueous droplets. These materials offer interesting possibilities for the controlled release of chemical species initially entrapped in the internal droplets. A better understanding of the stability conditions and release properties in double emulsions requires the use of model systems with a well-defined droplet size. In this paper, we use quasi-monodisperse double emulsions made of calibrated water droplets and oil globules to investigate the two mechanisms that are responsible for the release of a chemical substance (NaCl). (i) One is due to the coalescence of the thin liquid film separating the internal droplets and the globule surfaces. (ii) The other mechanism termed as 'compositional ripening' occurs without film rupturing; instead it occurs by diffusion and/or permeation of the chemical substance across the oil phase. By varying the proportions and/or the chemical nature of the surface active species it is possible to shift from one mechanism to the other one. We therefore study separately both mechanisms and we establish some basic rules that govern the behavior of W/O/W double emulsions. PMID- 11853932 TI - Sustained-delivery of an apolipoprotein E-peptidomimetic using multivesicular liposomes lowers serum cholesterol levels. AB - Receptor-mediated removal of lipoproteins by the liver is predominantly mediated by apolipoproteinE (apoE). Recent data show that a novel dual domain peptide (hE 18A) containing the 10-residue receptor-binding domain of human apoE, and a model class A amphipathic helix (18A), can associate with low density and very low density lipoproteins (LDL and VLDL) and enhance their uptake and degradation by HepG2 cells in vitro, and further, causes a dramatic reduction in plasma cholesterol levels in apoE-null mice. The in vivo cholesterol lowering effect, however, is short-lived because of rapid clearance of the peptide from the circulation. These results indicate that a therapeutic benefit may be achieved by sustained-delivery of this novel peptide as an alternate form of treatment for hypercholesterolemia and hypertriglyceridemia. In this report we describe the encapsulation of this peptide in a multivesicular liposome (MVL) depot-delivery system (DepoFoam). An in vitro plasma release assay showed sustained release of the peptide from the DepoFoam particles and its subsequent association with LDL and VLDL. Furthermore, a single subcutaneous dose of the DepohE-18A formulation in apoE-null mice, yielded a mean, cumulative 18% decrease in serum cholesterol levels after 6 days, and the cholesterol levels remained low at 8 days. Whereas, an equivalent dose of free peptide showed maximal cholesterol decrease by 4 h, followed by a rapid decline in efficacy by 24-48 h. Fractionation of the different lipoprotein fractions from serum showed that the majority of the serum cholesterol decrease was associated with the VLDL fraction, followed by LDL. These results indicate that the apoE peptidomimetic encapsulated in DepoFoam has potential as an alternative therapeutic treatment of hyperlipidemia. PMID- 11853933 TI - In vivo assessment of skin electroporation using square wave pulses. AB - The application of short-duration high-voltage pulses to the skin has been shown to enhance transdermal drug delivery by several orders of magnitude and to transiently permeabilize cells in tissue. Both exponentially decaying (ED) pulses and square wave (SW) pulses have been applied. The latter have also been used for electrochemotherapy. To date, their effect on skin integrity has not been analyzed. The scope of this work was (i) to investigate the effect induced by SW pulses on the stratum corneum and the skin, (ii) to evaluate the safety issue associated with electroporation, (iii) to contribute to the understanding of drug transport. Biophysical techniques (transepidermal water loss, chromametry, impedance and laser Doppler velocimetry or imaging measurement) and histological methods were combined to provide a global picture of the effects. Ten SW pulses applied to the skin induced a mild impairment of the skin barrier function and a dramatic decrease in skin resistance. These changes were reversible. A transient decrease (<5 min) in blood flow was observed. Neither inflammation, nor necroses were observed. These studies confirm the tolerance of the skin to square wave pulses in vivo. PMID- 11853935 TI - Development of a novel nasal nicotine formulation comprising an optimal pulsatile and sustained plasma nicotine profile for smoking cessation. AB - A novel nasal formulation, in the form of a nicotine-Amberlite resin complex powder has been developed that provided an optimal combined pulsatile and sustained plasma nicotine profile for smoking cessation. The adsorption isotherms of nicotine hydrogen tartrate salt on two types of Amberlite resins (IRP69 and IR120) were evaluated and the subsequent in vitro release properties of nicotine from the nicotine-Amberlite complex powders were tested using a Franz diffusion cell. Amberlite IRP69 and Amberlite IR120 are similar cationic exchange materials with the same ion-exchange capacity but due to a smaller particle size range (10 150 microm) Amberlite IRP69 had a better flow property and a better adsorptive capacity than Amberlite IR120. The material is used as an excipient in marketed pharmaceutical formulations. The highly water soluble salt, nicotine hydrogen tartrate, displayed good adsorption onto both types of Amberlite resin. The maximum adsorption of nicotine onto Amberlite IRP69 was 1.071 mg drug per mg resin. The cumulative release of drug from nicotine hydrogen tartrate-Amberlite complex powders showed that the higher the drug loading, the faster was the rate of release of the drug. Based on these results, various nicotine hydrogen tartrate-Amberlite IRP69 powder formulations containing different ratios of free to bound drug (50% to 100% bound) and a control solution were prepared and evaluated in a sheep model by nasal administration. The nicotine plasma profiles demonstrated that an initial rapid peak plasma level of nicotine followed by a sustained elevated level could be achieved by adjusting the ratio of free to bound nicotine in the Amberlite powder formulation. The curves obtained from some of the formulations were comparable to those predicted from a computer-generated pharmacokinetic model. PMID- 11853934 TI - Enhanced tumor cell selectivity of adriamycin-monoclonal antibody conjugate via a poly(ethylene glycol)-based cleavable linker. AB - A novel linker consisting of poly(ethylene glycol) (PEG) and dipeptide was used for conjugation of adriamycin with tumor-specific monoclonal antibody, NL-1, to confirm that the linker can be cleaved selectively with the tumor specific enzyme to express cytotoxicity of the anti-tumor agent. Initially, adriamycin-conjugated PEG linkers through different amino acid compositions, alanyl-valine (Ala-Val), alanyl-proline (Ala-Pro), and glycyl-proline (Gly-Pro) sequences, were prepared to confirm selective digestion with model enzymes. Adriamycin was released by particular model endoproteases, thermolysin and proline endopeptidase, from the linkers with different efficiency. When conjugates were prepared using these adriamycin-bound linkers, conjugates had no loss of binding affinity and specificity for common acute lymphoblastic leukemia antigen (CALLA) expressed on the Daudi cell surfaces as the target of NL-1 antibody. In addition, adriamycin release from the conjugates was also confirmed by incubating them with specific proteases. Tumor cell growth was inhibited dose-dependently for the conjugates carrying Ala-Val and Gly-Pro linkers, whereas significant inhibitory effect was abolished for the conjugate carrying Ala-Pro linker, indicating that cytotoxic effect can be controlled by specificity of antibody and composition of linker peptide. IC(50) for Ala-Val linked conjugate was approximately 3.5 microg/ml and that for Gly-Pro linked conjugate was 5.2 microg/ml. PEG-dipeptidyl linker demonstrated here will be an effective tool for the preparation of immunoconjugate, especially specific activation of anti-tumor agents at desired tumor tissues. PMID- 11853936 TI - Optimization of factors influencing the transfection efficiency of folate-PEG folate-graft-polyethylenimine. AB - Folate-poly(ethylene glycol)-folate-grafted-polyethylenimine (FPF-g-PEI) was synthesized over a range of grafting ratios of folate-poly(ethylene glycol) folate (FPF) to polyethylenimine (PEI). The conjugation was determined using the absorbance at 363 nm for each polymer. FPF-g-PEIs were determined to have 2.3, 5.2, 9.3 and 20 FPF linear polymers grafted to each PEI. The average molecular weight was calculated to be approximately 34,848, 47,266, 64,823 and 110,640 Da, respectively. The pH profiles of FPF-g-PEIs suggest that the polymers have endosomal disruption capacity, and the gel electrophoretic band retardation showed efficient condensation of DNA. The transfection efficiency, determined using plasmid encoding luciferase, was dependent on the cell type and was different for CT-26 colon adenocarcinoma, KB oral epidermoid, and normal smooth muscle cells (SMC). The relative toxicity of polymer/plasmid complexes was determined using the MTT colorimetric assay. At neutral charge ratio, FPF-g PEI/pLuc complexes were less toxic to cells and showed higher transfection in cancer cells compared to PEI/pLuc complexes. Smooth muscle cells showed no specificity for FPF-g-PEI/pLuc complexes, whereas PEI/pLuc complexes showed a higher transfection efficiency. The transfection efficiency increased when neutral polymer/DNA complex concentrations increased, but decreased when positively charged polymer/DNA complex concentrations increased. There was little increase in toxicity when FPF-5.2g-PEI/pLuc complex concentrations increased. PMID- 11853937 TI - A new synthesis of galactose-poly(ethylene glycol)-polyethylenimine for gene delivery to hepatocytes. AB - A synthesis method of conjugating polyethylenimine (PEI) derivatives with terminally galactose-grafted poly(ethylene glycol) (PEG) was developed by using a bifunctional PEG derivative containing both an alpha-vinyl sulfone (VS) and an omega-N-hydroxysuccinimidyl (NHS) ester groups (VS-PEG-NHS). VS-PEG-NHS is commonly used as a crosslinker to modify proteins with ligands by first coupling amine groups to the NHS ester, followed by coupling sulfhydryl groups to the VS ester, because the reaction of VS groups with amine groups of proteins is suppressed below pH 8. However, the 1H-NMR determination of the conjugated products of branched PEI (M(w)=25 kDa) with VS-PEG-NHS at pH 6.0-8.0 indicated that the VS groups were completely bound to the amine groups of PEI as well as the NHS groups. At pH 7.0, all VS groups reacted with the primary, secondary, or tertiary amine groups of PEI in 2 h. Such different reaction behaviors would be due to a higher density of amine groups of PEI as compared with those of proteins. In contrast, the reactions with a small molecular monoamine, such as p aminophenyl beta-D-galactopyranoside, showed that the NHS groups selectively coupled with the amine groups, and the VS groups remained completely intact. The NHS groups of VS-PEG-NHS were selectively conjugated to amine groups of p aminophenyl beta-D-galactopyranoside (VS-PEG-Gal). Then, the VS groups of Gal-PEG unit were completely conjugated with the primary, secondary, or tertiary amine groups of PEI. Thus, the use of only two reaction steps could conveniently carry out the conjugation of terminally galactose-grafted PEG to 1 and 5 mol.% of amine functions in PEI. The gel retardation assay of the complexes between Gal-PEG-PEI and plasmid DNA indicated that these polymeric gene carriers possess the potent ability to condense plasmid DNA electrostatically as well as PEI. The transfection efficiency with 1% Gal-PEG-PEI in human hepatocyte-derived cell lines (HepG2), a model of parenchymal cells in liver (hepatocytes), was superior to that of PEI at their corresponding optimal ratios of polymer to plasmid DNA. In HepG2 cells, luciferase activity with 1% Gal-PEG-PEI at an N/P ratio of 20 was 2.1-fold greater than that of PEI at an N/P ratio of 5. In mouse fibroblasts (NIH3T3) that have no ASGP receptors, the transfection efficiency with 1% Gal-PEG PEI drastically decreased to 1/40 of that with PEI. These data indicate that a new synthesis method can produce polyethylenimine derivatives with terminally galactose-grafted poly(ethylene glycol) for specific gene targeting to the liver. PMID- 11853938 TI - PEG grafted polylysine with fusogenic peptide for gene delivery: high transfection efficiency with low cytotoxicity. AB - For efficient gene delivery into cells, a new formulation method based on using polyethylene glycol (PEG) grafted poly(L-lysine) (PLL) and a fusogenic peptide is presented in this study. First, PEG grafted PLL (PEG-g-PLL) was complexed with DNA by controlling the polymer/DNA ratio to form negatively charged nano particulate complexes. A positively charged fusogenic peptide, KALA, was then coated by ionic interaction onto the surface of polymer/DNA complexes to make net positively charged KALA/polymer/DNA complexes. The use of PEG-g-PLL for KALA coating significantly suppressed the aggregation of complexes due to steric stabilization effect of PEG present on the surface, while the use of PLL alone induced severe aggregation of the complexes via KALA mediated inter-particulate cross-linking. For PEG-g-PLL/DNA complexes, enhanced transfection efficiency was observed with increasing amount of KALA. This suggests that maintaining the size of DNA/polymer complexes after KALA coating plays an important role in gene transfection. KALA/DNA/PEG-g-PLL complexes exhibited lower cytotoxicity compared with other polymer/DNA complexes. PMID- 11853939 TI - Multipulse drug permeation across a membrane driven by a chemical pH-oscillator. AB - Chemical pH-oscillators provide a potential means for modulating the delivery of acidic or basic drugs across lipophilic membranes. Previous attempts to exploit this notion were not completely successful due to quenching of the pH oscillator in the presence of the drug, most likely by a buffering mechanism. Here we show that multiple, periodic pulses of drug flux across a membrane can be achieved when the concentration of drug is sufficiently low. Advantage is taken of a pH oscillator system whose periodicity is slower than that of previously considered oscillators. PMID- 11853940 TI - Chemical hydrolysis of DOTAP and DOPE in a liposomal environment. AB - In this study the hydrolysis kinetics of 1,2-dipalmitoyl-sn-glycero-3 phosphocholine (DPPC), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) and 1,2-dioleoyltrimethylammoniumpropane (DOTAP) in net neutral DPPC-DOPE (3:1, mol/mol) and cationic DOTAP-DOPE (1:1, mol/mol) liposomes are described. The log k(obs)-pH profile for DOTAP-DOPE liposomes differs markedly from earlier observed hydrolysis profiles: the slope approaches zero in the acidic region and +1 in the alkaline region. The concept of amine-influenced hydrolysis is introduced to explain the lack of pH dependency in the acidic region of the log k(obs)-pH profiles. PMID- 11853941 TI - Free knot splines for biochemical data. AB - The use of spline functions in the analysis of empirical two-dimensional (2-D) data (y(i), x(i)) is described. Spline functions are excellent empirical functions, which can be used with advantage instead of other ones, such as polynomials or exponentials. The knot location seen as variable value corresponds to classical parameter used to describe oxidation curves. An application on characterization of LDL oxidability shows free knot splines in a regression context. PMID- 11853942 TI - Evaluation of two cortical fraction estimation algorithms for the calculation of dynamic magnetic resonance renograms. AB - With the high resolution of dynamic magnetic resonance imaging (MRI) scans it is possible to measure cortical renograms directly, but due to partial volume effects this is impossible for medullary renograms. With weighted subtraction of the cortical renogram from a mixed renogram it becomes possible to extract the medullary renogram. For this subtraction the fraction of cortical tissue, present in the region of interest in which the mixed renogram is determined, has to be calculated. We have evaluated two algorithms for calculation of the cortical fraction. Both algorithms use the fact that during an interval after the start of the cortical enhancement no medullary enhancement occurs. One algorithm calculates the ratio between the slopes of both enhancement curves. The other is based on minimising the medullary signal values using a least squares error (LSE) method. Using a computer model of the renograms and measurements on real patients we analysed the accuracy of both methods and determined the best parameters for each. PMID- 11853943 TI - Using contexts and R-R interval estimation in lossless ECG compression. AB - The paper presents a new lossless ECG compression scheme. The short-term predictor and the coder use conditioning on a small number of contexts. The long term prediction is based on an algorithm for R-R interval estimation. Several QRS detection algorithms are investigated to select a low complexity and reliable detection algorithm. The coding of prediction residuals uses primarily the Golomb Rice (GR) codes, but, to improve the coding results, escape codes GR-ESC are used in some contexts for a limited number of samples. Experimental results indicate the good overall performance of the lossless ECG compression algorithms (reducing the storage needs from 12 to about 3-4 bits per sample). The scheme consistently outperforms other waveform or general purpose coding algorithms. PMID- 11853944 TI - Relationship between Brier score and area under the binormal ROC curve. AB - If we consider the Brier score (B) in the context of the signal detection theory and assume that it makes sense to consider the existence of B as a parameter for the population (let B be this B), and if we assume that the calibration in the observer's probability estimate is perfect, we find that there is a theoretical relationship between B and the area under the binormal receiver operating characteristic (ROC) curve, A(Z). We have derived this theoretical functional relationship between B and A(Z), by using the parameter a and b in the binormal ROC model and the prior probability of signal events (alpha); here, the two underlying normal distributions are N and N; and, a= and b=. We empirically found that, if parameters b and alpha are constant, B values in relation to given A(Z) values monotonically decrease as A(Z) values increase, and these relationship curves have monotonically decreasing slopes. PMID- 11853946 TI - A SAS macro for the analysis of cell survival curves. AB - This paper describes a SAS macro for the statistical analyses of cell survival data obtained after radiation treatment using the methods of R.E. Tarone et al. (Mutation Research 111 (1983) 79-96). These analyses are usually required on a routine basis by all biomedical research laboratories involved in cell survival assays generating dose-response curves aimed at characterizing radiosensitive mutant cell strains or individuals whose body cells exhibit enhanced sensitivity to radiation and other genotoxic agents. Statistical methods of linear regression are applied to data from repeated experiments with a cell line/strain and weighted estimates of a common slope and its variance are obtained. The methods are currently implemented in two APL programs. These programs are not easily accessible to most biomedical statisticians and researchers because APL is not a common software tool for statistical analysis. Implementation of these methods in SAS, a widely used commercial software for statistical analysis, is expected to help resolve this issue. We illustrate the application of the macro using an example data set obtained in our laboratory, and hope that other investigators may find it useful in analyzing their data. PMID- 11853945 TI - A new WIN32 computer program for estimating immunoassay variance functions. AB - A new WIN32 computer program for estimating immunoassay variance functions is described. It replaces and enhances an MS-DOS version written more than a decade ago. Input data can consist of either runs of raw replicated measurements (maximum: 40,000 observations) or sets of values of N, mean and SD (maximum: 20,000 sets). Data can be imported from Paradox, dBase, Foxpro or Access database tables, from text files, or via the Windows clipboard. The program produces histogram plots of sets of replicates and plots containing up to 12 variance functions expressed in terms of variance, SD or CV(%) versus the mean (imprecision profiles). Any specified view of a variance function graph can be inset into any other view. Interactive point and click design dialogues give virtually complete control over graph appearance. Graphs can be exported to external applications via the clipboard or disk files, in either bitmap or Windows enhanced metafile format. PMID- 11853948 TI - SAS and SPLUS programs to perform Cox regression without convergence problems. AB - When analyzing survival data, the parameter estimates and consequently the relative risk estimates of a Cox model sometimes do not converge to finite values. This phenomenon is due to special conditions in a data set and is known as 'monotone likelihood'. Statistical software packages for Cox regression using the maximum likelihood method cannot appropriately deal with this problem. A new procedure to solve the problem has been proposed by G. Heinze, M. Schemper, A solution to the problem of monotone likelihood in Cox regression, Biometrics 57 (2001). It has been shown that unlike the standard maximum likelihood method, this method always leads to finite parameter estimates. We developed a SAS macro and an SPLUS library to make this method available from within one of these widely used statistical software packages. Our programs are also capable of performing interval estimation based on profile penalized log likelihood (PPL) and of plotting the PPL function as was suggested by G. Heinze, M. Schemper, A solution to the problem of monotone likelihood in Cox regression, Biometrics 57 (2001). PMID- 11853947 TI - Surveyance of disease frequency in a population by linkage to diagnostic laboratory databases. A system for monitoring the incidences of hyper- and hypothyroidism as part of the Danish iodine supplementation program. AB - In Denmark an increase in iodine intake through salt iodization has been introduced in 1998. In parallel a program for surveyance of thyroid diseases in the population was developed as recommended by UNICEF and WHO. OBJECTIVE: To develop and evaluate a computer based system to identify and register new cases of hyper- and hypothyroidism in a well defined cohort, by linkage to diagnostic laboratory databases. DESIGN AND RESULTS: (1) Two sub cohorts for monitoring were defined (n=535,859), and evaluated to minimize loss of new cases. Collaboration was established with laboratories covering thyroid hormone analyses in the cohort; (2) a diagnostic algorithm was defined and evaluated against clinical practice; (3) evaluation of the laboratory methods employed by the four participating laboratories, to ensure they would reach the same diagnosis in a patient; (4) a register database was developed which used data imported from the laboratory databases to automatically identify previously unknown cases of hyper- and hypothyroidism and record diagnostic activity in the area. All parts of the registration were carefully evaluated. CONCLUSION: We describe for the first time a computer based system for prospective measuring the incidence rate of hyper- and hypothyroidism. The system is particularly useful for monitoring of iodine supplementation programmes. PMID- 11853949 TI - Software for temporal gait data analysis. AB - This study presents a computer program, developed to support a low-cost, portable telemetry system that has been designed to assess footfall timing. This software frees the user from data processing and allows concentration on data analysis. The new technique has been applied with accuracy and reliability to the analysis of the gait of orthopedic patients, athletes, mountaineers, etc. The subroutines developed for data acquisition, storage and analysis are explained in detail, and an example is presented. PMID- 11853950 TI - Computer-assisted preoperative planning of a novel design of total ankle replacement. AB - Computer-assisted preoperative planning are particularly sought for enhancing surgical implantation and improving clinical outcome of ankle joint replacement arthroplasty. A planning tool was designed to enhance the surgical implantation of a novel ligament-compatible three-component prosthesis. The tool includes a geometric and mechanical model of the ankle complex. The geometry of the articular surfaces and of main ligaments is defined from digitisation on a scanned lateral radiographs of the joint. From the expected movement arc and from component dimensions, a mechanical model of the ligaments is used to select the optimal size and the position the three prosthesis components. The planning is based on the minimisation of ligament change in length and of bone stock to be removed. The main output report is a detailed picture of the replaced ankle with indications of component locations and quantitative measure of bone cuts. An advanced report also superimposes the resulting prediction of prosthesis component kinematics at the replaced joint to the original radiographs. PMID- 11853951 TI - Ascorbic acid: much more than just an antioxidant. AB - Vitamin C (ascorbic acid (AA)) is very popular for its antioxidant properties. Consequently, many other important aspects of this multifaceted molecule are often underestimated or even ignored. In the present paper, we have tried to bring to the foreground some of these aspects, including the peculiarities of the AA biosynthetic pathway in different organisms, the remarkable function of AA as a co-substrate of many important dioxygenases, the role of AA-regenerating enzymes and the known pathways of AA catabolism, as well as the intriguing function of AA in gene expression. PMID- 11853952 TI - Interactions of wheat-germ agglutinin with GlcNAc beta 1,6Gal sequence. AB - The interactions of wheat-germ agglutinin (WGA) with the GlcNAc beta 1,6Gal sequence, a characteristic component of branched poly-N-acetyllactosaminoglycans, were investigated using isothermal titration calorimetry and X-ray crystallography. GlcNAc beta 1,6Gal exhibited an affinity greater than GlcNAc beta 1,4GlcNAc to all WGA isolectins, whereas Gal beta 1,6GlcNAc showed much less affinity than GlcNAc beta 1,4GlcNAc. X-ray structural analyses of the glutaraldehyde-crosslinked WGA isolectin 3 crystals in complex with GlcNAc beta 1,6Gal, GlcNAc beta 1,4GlcNAc and GlcNAc beta 1,6Gal beta 1,4Glc were performed at 2.4, 2.2 and 2.2 A resolution, respectively. In spite of different glycosidic linkages, GlcNAc beta 1,6Gal and GlcNAc beta 1,4GlcNAc exhibited basically similar binding modes to each other, in contact with side chains of two aromatic residues, Tyr64 and His66. However, the conformations of the ligands in the two primary binding sites were not always identical. GlcNAc beta 1,6Gal showed more extensive variation in the parameters defining the glycosidic linkage structure compared to GlcNAc beta 1,4GlcNAc, demonstrating large conformational flexibility of the former ligand in the interaction with WGA. The difference in the ligand binding conformation was accompanied by alterations of the side chain conformation of the amino acid residues involved in the interactions. The hydrogen bond between Ser62 and the non-reducing end GlcNAc was always observed regardless of the ligand type, indicating the key role of this interaction. In addition to the hydrogen bonding and van der Waals interactions, CH--pi interactions involving Tyr64, His66 and Tyr73 are suggested to play an essential role in determining the ligand binding conformation in all complexes. One of the GlcNAc beta 1,6Gal ligands had no crystal packing contact with another WGA molecule, therefore the conformation might be more relevant to the interaction mode in solution. PMID- 11853953 TI - Real-time kinetic analysis of hCG-monoclonal antibody interaction using radiolabeled hCG probe: presence of two forms of Ag-mAb complex as revealed by solid phase dissociation studies. AB - Real-time kinetics of ligand-ligate interaction has predominantly been studied by either fluorescence or surface plasmon resonance based methods. Almost all such studies are based on association between the ligand and the ligate. This paper reports our analysis of dissociation data of monoclonal antibody-antigen (hCG) system using radio-iodinated hCG as a probe and nitrocellulose as a solid support to immobilize mAb. The data was analyzed quantitatively for a one-step and a two step model. The data fits well into the two-step model. We also found that a fraction of what is bound is non-dissociable (tight-binding portion (TBP)). The TBP was neither an artifact of immobilization nor does it interfere with analysis. It was present when the reaction was carried out in homogeneous solution in liquid phase. The rate constants obtained from the two methods were comparable. The work reported here shows that real-time kinetics of other ligand ligate interaction can be studied using nitrocellulose as a solid support. PMID- 11853954 TI - Degradation of non-esterified and esterified xanthophylls by free radicals. AB - beta-Carotene and other xanthophylls present in pepper fruit as both free and esterified forms were oxidized using a free radical initiator (2,2'-azo-bis isobutyronitrile). Capsorubin was degraded most slowly, followed by zeaxanthin, capsanthin, and beta-carotene. The presence of keto groups at the ends of the polyene chain could be a structural factor contributing to this difference in reactivity. It was also shown that whereas capsanthin and its esters and capsorubin and its esters were degraded at the same rate, zeaxanthin esters responded differently to the oxidation process, and were degraded more quickly than free zeaxanthin. The presence of unsaturated fatty acids (mainly linoleic) that esterify zeaxanthin help to accelerate the degradation of this xanthophyll and decreasing its antioxidant action. The antioxidant capacity of capsorubin and capsanthin (both in free and esterified form) exclusive to the genus capsicum should be taken into account. PMID- 11853955 TI - Kinetic analysis and mechanistic aspects of autoxidation of catechins. AB - A peroxidase-based bioelectrochemical sensor of hydrogen peroxide (H(2)O(2)) and a Clark-type oxygen electrode were applied to continuous monitoring and kinetic analysis of the autoxidation of catechins. Four major catechins in green tea, (-) epicatechin, (-)-epicatechin gallate, (-)-epigallocatechin, and (-) epigallocatechin gallate, were used as model compounds. It was found that dioxygen (O(2)) is quantitatively reduced to H(2)O(2). The initial rate of autoxidation is suppressed by superoxide dismutase and H(+), but is independent of buffer capacity. Based on these results, a mechanism of autoxidation is proposed; the initial step is the one-electron oxidation of the B ring of catechins by O(2) to generate a superoxide anion (O(2)(*-)) and a semiquinone radical, as supported in part by electron spin resonance measurements. O(2)(*-) works as a stronger one-electron oxidant than O(2) against catechins and is reduced to H(2)O(2). The semiquinone radical is more susceptible to oxidation with O(2) than fully reduced catechins. The autoxidation rate increases with pH. This behavior can be interpreted in terms of the increase in the stability of O(2)(*-) and the semiquinone radical with increasing pH, rather than the acid dissociation of phenolic groups. Cupric ion enhances autoxidation; most probably it functions as a catalyst of the initial oxidation step of catechins. The product cuprous ion can trigger a Fenton reaction to generate hydroxyl radical. On the other hand, borate ion suppresses autoxidation drastically, due to the strong complex formation with catechins. The biological significance of autoxidation and its effectors are also discussed. PMID- 11853956 TI - Muscarinic receptor subtypes mediate stimulatory and paradoxical inhibitory effects on an insulin-secreting beta cell line. AB - Acetylcholine (ACh), a major neurotransmitter from the autonomic nervous system, regulates the cholinergic stimulation of insulin secretion, through interactions with muscarinic receptors. The present study has characterised the individual involvement of muscarinic receptor subtypes in ACh-induced insulin secretion, using clonal beta cells and selective muscarinic receptor antagonists. BRIN BD11 cells clearly expressed mRNA encoding m1--m4 whereas m5 was not detected by RT PCR. Insulin release was measured from BRIN BD11 cells treated with ACh in the presence of muscarinic receptor antagonists at concentrations ranging from 3 nM to 1 microM. 300 nM of muscarinic toxin-3 (M4 antagonist) and 1 microM of methoctramine (M2 antagonist) increased ACh (100 microM) stimulated insulin secretion by 168% and 50% respectively (ANOVA, P<0.05). The antagonists alone had no effect on insulin secretion. In contrast, 300 nM of pirenzepine (M1 antagonist) and 30 nM of hexahydro-sila-difenidol p-fluorohydrochloride (M3 antagonist) inhibited ACh stimulation by 91% and 84% respectively (ANOVA, P<0.01). It is concluded that ACh acts on different receptor subtypes producing both a stimulatory and an inhibitory action on insulin release. PMID- 11853957 TI - Vascular reactions to in vivo electroporation: characterization and consequences for drug and gene delivery. AB - In vivo electroporation (EP) is gaining momentum for drug and gene delivery. In particular, DNA transfer by EP to muscle tissue can lead to highly efficient long term gene expression. We characterized a vascular effect of in vivo EP and its consequences for drug and gene delivery. Pulses of 10-20,000 micros and 0.1-1.6 kV/cm were applied over hind- and forelimb of mice and perfusion was examined by dye injection. The role of a sympathetically mediated vasoconstrictory reflex was investigated by pretreatment with reserpine. Expression of a transferred gene (luciferase), permeabilization (determined using (51)Cr-EDTA), membrane resealing and effects on perfusion were compared to assess the significance of the vascular effects. Above the permeabilization threshold, a sympathetically mediated Raynaud like phenomenon with perfusion delays of 1-2 min was observed. Resolution of this phase followed kinetics of membrane resealing. Above a second threshold, irreversible permeabilization led to long perfusion delays. These vascular reactions (1) affect kinetics of drug delivery, (2) predict efficient DNA transfer, which is optimal during short perfusion delays, and (3) might explain electrocardiographic ST segment depressions after defibrillation as being caused by vascular effects of EP of cardiac muscle. PMID- 11853958 TI - Tissue-specific regulation of Ca(2+) channel protein expression by sex hormones. AB - The L-type Ca(2+) channel pore-forming alpha subunit, alpha(1C) can be detected in brain and heart as two proteins with molecular masses of approximately 240 kDa and approximately 190 kDa known as alpha(1C-long) and alpha(1C-short), respectively. In brain, the alpha(1C-short) is thought to be the product of a approximately 50 kDa C-terminus calpain-mediated proteolytic deletion. We now show that uterine smooth muscle also possesses alpha(1C-long) and alpha(1C-short) isoforms, and that the relative expression of these two forms is regulated by sex hormones in a tissue-specific manner. Protein expression of alpha(1C) L-type Ca(2+) channels was examined in uterine smooth muscle, brain and heart, comparing non-pregnant (NP) estrus vs. late-pregnant (21 days) rats. The two forms of alpha(1C) were detected in all studied tissues. In late-pregnant uterus, alpha(1C long) doubled the expression of alpha(1C-short); in NP uterus the opposite occurred. However, these changes were restricted to the uterine muscle, with no changes in brain and heart. To investigate the mechanism of such regulation, ovariectomized rats were treated with sex hormones, progesterone (P4) and/or 17beta-estradiol (estrogen, E2). P4 treatment, which yielded P4 plasma levels of 5 +/- 1 ng/ml and a high P4/E2 ratio (3 +/- 1.5 x 10(3)) similar to the ratio in late-pregnant uterus (1.5 +/- 0.3 x10(3)), facilitated alpha(1C-long) expression. In contrast, E2 or E2+P4 treatment that increased E2 plasma levels to 60 +/- 8 pg/ml and 75 +/- 24 pg/ml, produced low P4/E2 ratios of 0.03 +/- 0.006 and 0.2 +/ 0.1, respectively. These low P4/E2 ratios also found in NP rats at estrus (0.3 +/- 0.1) favored the expression of alpha(1C-short) form in myometrium. Neither hormone treatment altered alpha(1C) expression in brain or heart. Our results indicate that expression of alpha(1C) isoforms depends on P4/E2 ratios. Plasma P4/E2 ratios <1 x 10(3) favor the expression of the alpha(1C-short); whereas ratios >1 x 10(3) facilitate the expression of the alpha(1C-long) form. This regulation is tissue-specific for myometrium since it did not occur in heart and brain tissues. PMID- 11853959 TI - Redox regulation of cytosolic glycerol-3-phosphate dehydrogenase: Cys(102) is the target of the redox control and essential for the catalytic activity. AB - Cytosolic glycerol-3-phosphate dehydrogenase (cG3PDH) occupies the branch point between the glycolytic pathway and triglyceride biosynthesis. However, the regulatory mechanism of the cG3PDH activity has remained obscure. Here we report that cG3PDH is efficiently inhibited by modification of the thiol group through a redox mechanism. In this study, we found that sodium selenite and nitric oxide (NO) donors such as S-nitroso-N-acetylpenicillamine and 3-morpholinosydnonimine inhibited cG3PDH activity, and that similar effects could be achieved with selenium metabolites such as selenocysteine and selenomethionine. Furthermore, we found that reducing agents, such as dithiothreitol and beta-mercaptoethanol, restored the cG3PDH activity suppressed by selenite and NO both in vitro and in cultured cells. Buthionine sulfoximine depleted levels of both reduced glutathione and the oxidized form but had no effect on the suppression of cG3PDH activity by selenite in cultured cells. Moreover, thiol-reactive agents, such as N-ethylmaleimide and o-iodosobenzoic acid, blocked the enzyme activity of cG3PDH through the modification of redox-sensitive cysteine residues in cG3PDH. The inhibitor of NO synthase, L-N(G)-nitro-arginine, restored the cG3PDH activity inhibited by NO in cultured cells, whereas the inhibitor of guanylyl cyclase, 1H [1,2,4] oxadiazole[4,3-alpha] quinoxalin-1-one (ODQ), has no effect. NO directly inhibits cG3PDH activity not via a cGMP-dependent mechanism. Finally, using site directed mutagenesis, we found that Cys(102) of cG3PDH was sensitive to both selenite and NO. From the results, we suggest that cG3PDH is a target of cellular redox regulation. PMID- 11853960 TI - Evaluation of glycated glucagon-like peptide-1(7-36)amide in intestinal tissue of normal and diabetic animal models. AB - Glucagon-like peptide-1(7-36)amide (tGLP-1) is an important insulin-releasing hormone of the enteroinsular axis which is secreted by endocrine L-cells of the small intestine following nutrient ingestion. The present study has evaluated tGLP-1 in the intestines of normal and diabetic animal models and estimated the proportion present in glycated form. Total immunoreactive tGLP-1 levels in the intestines of hyperglycaemic hydrocortisone-treated rats, streptozotocin-treated mice and ob/ob mice were similar to age-matched controls. Affinity chromatographic separation of glycated and non-glycated proteins in intestinal extracts followed by radioimmunoassay using a fully cross-reacting anti-serum demonstrated the presence of glycated tGLP-1 within the intestinal extracts of all control animals (approximately 19% of total tGLP-1 content). Chemically induced and spontaneous animal models of diabetes were found to possess significantly greater levels of glycated tGLP-1 than controls, corresponding to between 24--71% of the total content. These observations suggest that glycated tGLP-1 may be of physiological significance given that such N-terminal modification confers resistance to DPP IV inactivation and degradation, extending the very short half-life (<3 min) and bioactivity of the native peptide. PMID- 11853961 TI - Glutathione transferase isoenzymes from frog (Xenopus laevis) liver and embryo. AB - The expression of glutathione transferase isoenzymes has been investigated in embryo and adult liver of the frog Xenopus laevis. By analysing the GST isoenzymes recovered from GSH-affinity chromatography in terms of electrophoretic mobility, HPLC elution profile, immunological reactivity, N-terminal amino acid sequence and mass spectrometry molecular mass no significant difference in the GST subunit composition between embryos and liver was found. In both tissues the same three subunits, showing similarity to mu, alpha and sigma class GSTs, are present. These results, together with those previously reported for toad (Bufo bufo), strongly support the notion that the transition from an aquatic environment to a terrestrial atmosphere containing high oxygen concentration has accompanied specific GST gene expression. PMID- 11853962 TI - Spatiotemporal dynamics of contact activation factors of blood coagulation. AB - A new in vitro model is proposed for studying the spatiotemporal distributions of activated clotting factors, in which clotting is activated in a thin layer of non stirred plasma supplemented with a fluorogenic substrate and is monitored by fluorescence from its cleavage product. Analysis of the spatiotemporal dynamics of factor XIa and kallikrein in glass-activated human plasma provides evidence that both contact factors remain restricted to the glass surface and possibly a narrow boundary zone (<0.1 mm). The kinetics of factor XIa and kallikrein studied by a new method (in non-stirred plasma) coincided with those studied fluorimetrically with full stirring: their concentrations rapidly rose for the first few minutes after activation and then slowly declined. Factor XI and prekallikrein activation is likely to be restricted by the limited number of sites available for binding to the surface. The maximum concentration of the active factors was estimated at 2 x 10(8) molecules per mm(2) at the glass surface (irrespective of stirring). At the plastic surface, this value was 15--30 times lower. PMID- 11853963 TI - Prolonged blood glucose reduction in mrp-2 deficient rats (GY/TR(-)) by the glucose-6-phosphate translocase inhibitor S 3025. AB - Chlorogenic acid derivatives are potent inhibitors of hepatic glucose production by inhibition of the glucose-6-phosphate translocase component of the hepatic glucose-6-phosphatase system. The pharmacological proof of concept was clearly demonstrated during i.v. infusion of potent derivatives (S 4048, S 3483) in rats. However, the blood glucose lowering effect of S 4048 after bolus i.v. injection lasted only 60-90 min. Plasma clearance of S 4048 was very high, and the parent compound was rapidly and efficiently excreted into the bile of Wistar and GY/TR( ) rats, indicating that mrp-2 was not involved in this hepatobiliary elimination process. About 72% of the total administered radioactivity appeared in the bile within 20 min after i.v. bolus injection of the radiolabeled analogue [(3)H]S 1743 in a Wistar rat. However, in GY/TR(-) rats the dicarboxylic analogue of S 4048, S 3025, was cleared from the plasma less rapidly than its parent compound and its biliary elimination was comparatively low. In contrast, S 3025 exhibited comparable pharmacokinetics and biliary elimination profile as S 4048 in Wistar rats, suggesting that biliary elimination of S 3025 is facilitated by mrp-2, functionally absent in GY/TR(-) rats. Targeting to mrp-2 resulted in a significantly prolonged reduction of blood glucose levels in GY/TR(-) rats after i.v. bolus administration of S 3025. PMID- 11853964 TI - Perferryl complex of nitric oxide synthase: role in secondary free radical formation. AB - Neuronal nitric oxide synthase (NOS I) has been shown to generate nitric oxide (NO*) and superoxide (O(2)*-)during enzymatic cycling, the ratio of each free radical is dependent upon the concentration of L-arginine. Using spin trapping and electron paramagnetic resonance (EPR) spectroscopy, we recently reported that NOS I can oxidize ethanol (EtOH) to alpha-hydroxyethyl radical (CH(3)*CHOH). We speculated that the perferryl complex of NOS, (NOS-[Fe(5+)[double bond]O](3+)) was responsible for the generation of CH(3)*CHOH. Using potassium monopersulfate (KHSO(5)) to oxidize the heme of NOS I to NOS-[Fe(5+)[double bond]O](3+), we were able to demonstrate that this perferryl complex can oxidize L-arginine to L citrulline and NO*. Even in the absence of L-arginine, EtOH was oxidized to CH(3)*CHOH by NOS-[Fe(5+)[double bond]O](3+). Sodium cyanide (NaCN), a heme blocker, inhibited the formation of CH(3)*CHOH by NOS. PMID- 11853965 TI - Isolation and characterization of matrix proteoglycans from human nasal cartilage. Compositional and structural comparison between normal and scoliotic tissues. AB - The content, types and the fine structures of proteoglycans (PGs) present in human normal nasal cartilage (HNNC) were investigated and compared with those in human scoliotic nasal cartilage (HSNC). Three PG types were identified in both HNNC and HSNC; the large-sized high buoyant density aggrecan, which is the predominant PG population, and the small-sized low buoyant density biglycan and decorin. HSNC contained a significantly higher amount of keratan sulfate (KS) rich aggrecan (30%) of smaller hydrodynamic size as compared to HNNC. The average molecular sizes (M(r)s) of aggecan-derived chondroitin sulfate (CS) chains in both HNNC and HSNC were identical (18 kDa), but they significantly differ in disaccharide composition, since CS isolated from HSNC contained higher proportions of 6-sulfated disaccharides as compared to those from HNNC. Scoliotic tissue contained also higher amounts (67%) of the small PGs, biglycan and decorin as compared to HNNC. It is worth noticing that both normal and scoliotic human nasal cartilage contain also non-glycanated forms of decorin and biglycan. Dermatan sulfate (DS) was the predominant glycosaminoglycan (GAG) present on biglycan and decorin in both tissues. The small PGs-derived CS chains in both normal and scoliotic cartilage had the same M(r) (20 kDa), whereas DS chains from scoliotic cartilage were of greater M(r) (32 kDa) than those from normal cartilage (24 kDa). Furthermore, scoliotic tissue-derived DS chains contained higher amounts of iduronate (20%) as compared to those of normal cartilage (12%). Disaccharide analysis of small PGs showed that both HNNC and HSNC were rich in 4 sulfated disaccharides and in each case, the small size PGs contained a considerably higher proportion of 4-sulfated disaccharides than the aggrecan of the same tissue. The higher amounts of matrix PGs identified in scoliotic tissue as well as the differences in fine chemical composition of their GAG chains may reflect the modified architecture and functional failure of scoliotic tissue. PMID- 11853966 TI - Binding of CaMKII to the giant muscle protein projectin: stimulation of CaMKII activity by projectin. AB - Projectin is an integral high molecular mass protein of insect flight muscle binding to myosin and paramyosin. Yet, the role of projectin in insect flight muscle is not well understood. In this study we provide evidence for the interaction of projectin with the calcium sensor Ca(2+)/calmodulin-dependent protein kinase II (CaMKII). Two CaMKII variants of 52 and 60 kDa, respectively, of locust flight muscle cells were shown by an anti-CaMKII antibody. Both variants were associated to myofibrils. The 52 kDa variant was also shown abundant in the cytosol. The cytosolic CaMKII variant was co-precipitated in vitro with externally added purified projectin in a dose-dependent manner. To specify the binding properties of CaMKII to projectin we used both purified projectin from the flight muscle of locust and CaMKII from rat forebrain, a naturally rich source of CaMKII. CaMKII is highly conserved even between insects and mammals. The binding of CaMKII to native projectin was demonstrated in vitro by the solid phase enzyme assay, immunoprecipitation, and 'overlay' binding. One mol projectin bound maximally 1.38 +/- 0.02 mol CaMKII in vitro with a K(d) of 3.08 Z+/- 0.09 nM. Application of in vitro autophosphorylated CaMKII revealed a decreased stoichiometry of binding to projectin (0.86 +/- 0.04 mol mol(-1)) accompanied by a lower affinity (K(d) of 5.54 +/- 0.73) compared to non autophosphorylated CaMKII. Furthermore, the CaMKII phosphotransferase activity was stimulated up to 2-fold by projectin. Even in the presence of calmodulin projectin enhanced the CaMKII activity moderately. Our data suggest that projectin represents a subcellular compartment for CaMKII to achieve its specificity, and activity in insect flight muscle cells. PMID- 11853967 TI - Differential response of fast and slow myosin ATPase from skeletal muscle to F actin and to phalloidin F-actin. AB - Fast muscle myosin responds in similar way to F-actin and to phalloidin F-actin. It is activated 7.5 fold at infinite F-actin concentration and 6.8 fold at infinite phalloidin F-actin. The actomyosin dissociation constants are 0.89 +/- 0.34 microM with F-actin and 0.90 +/- 0.71 microM with phalloidin F-actin. Slow muscle myosin responds differently to F-actin and to phalloidin F-actin. It is activated 3.76 fold at infinite F-actin concentration and only 2.27 fold at infinite phalloidin F-actin concentration. The actomyosin dissociation constants are 1.95 +/- 1.27 microM with F-actin and 0.27 +/- 0.16 microM with phalloidin F actin. At first glance this means that substitution of F-actin with phalloidin F actin magnifies the difference between fast muscle and slow muscle myosins. Furthermore the change of the dissociation constants may affect the contractile force of the attached crossbridge. PMID- 11853968 TI - A novel two-step extraction method with detergent/polymer systems for primary recovery of the fusion protein endoglucanase I-hydrophobin I. AB - Extraction systems for hydrophobically tagged proteins have been developed based on phase separation in aqueous solutions of non-ionic detergents and polymers. The systems have earlier only been applied for separation of membrane proteins. Here, we examine the partitioning and purification of the amphiphilic fusion protein endoglucanase I(core)-hydrophobin I (EGI(core)-HFBI) from culture filtrate originating from a Trichoderma reesei fermentation. The micelle extraction system was formed by mixing the non-ionic detergent Triton X-114 or Triton X-100 with the hydroxypropyl starch polymer, Reppal PES100. The detergent/polymer aqueous two-phase systems resulted in both better separation characteristics and increased robustness compared to cloud point extraction in a Triton X-114/water system. Separation and robustness were characterized for the parameters: temperature, protein and salt additions. In the Triton X-114/Reppal PES100 detergent/polymer system EGI(core)-HFBI strongly partitioned into the micelle-rich phase with a partition coefficient (K) of 15 and was separated from hydrophilic proteins, which preferably partitioned to the polymer phase. After the primary recovery step, EGI(core)-HFBI was quantitatively back-extracted (K(EGIcore-HFBI)=150, yield=99%) into a water phase. In this second step, ethylene oxide-propylene oxide (EOPO) copolymers were added to the micelle-rich phase and temperature-induced phase separation at 55 degrees C was performed. Total recovery of EGI(core)-HFBI after the two separation steps was 90% with a volume reduction of six times. For thermolabile proteins, the back-extraction temperature could be decreased to room temperature by using a hydrophobically modified EOPO copolymer, with slightly lower yield. The addition of thermoseparating co-polymer is a novel approach to remove detergent and effectively releases the fusion protein EGI(core)-HFBI into a water phase. PMID- 11853969 TI - A facilitated electron transfer of copper--zinc superoxide dismutase (SOD) based on a cysteine-bridged SOD electrode. AB - The direct electrochemical redox reaction of bovine erythrocyte copper--zinc superoxide dismutase (Cu(2)Zn(2)SOD) was clearly observed at a gold electrode modified with a self-assembled monolayer (SAM) of cysteine in phosphate buffer solution containing SOD, although its reaction could not be observed at the bare electrode. In this case, SOD was found to be stably confined on the SAM of cysteine and the redox response could be observed even when the cysteine-SAM electrode used in the SOD solution was transferred to the pure electrolyte solution containing no SOD, suggesting the permanent binding of SOD via the SAM of cysteine on the electrode surface. The electrode reaction of the SOD confined on the cysteine-SAM electrode was found to be quasi-reversible with the formal potential of 65 +/- 3 mV vs. Ag/AgCl and its kinetic parameters were estimated: the electron transfer rate constant k(s) is 1.2 +/- 0.2 s(-1) and the anodic (alpha(a)) and cathodic (alpha(c)) transfer coefficients are 0.39 +/- 0.02 and 0.61 +/- 0.02, respectively. The assignment of the redox peak of SOD at the cysteine-SAM modified electrode could be sufficiently carried out using the native SOD (Cu(2)Zn(2)SOD), its Cu- or Zn-free derivatives (E(2)Zn(2)SOD and Cu(2)E(2)SOD, E designates an empty site) and the SOD reconstituted from E(2)Zn(2)SOD and Cu(2+). The Cu complex moiety, the active site for the enzymatic dismutation of the superoxide ion, was characterized to be also the electroactive site of SOD. In addition, we found that the SOD confined on the electrode can be expected to possess its inherent enzymatic activity for dismutation of the superoxide ion. PMID- 11853970 TI - The roles of ATP and calcium in morphological changes and cytotoxicity induced by 1,4-benzoquinone in platelets. AB - To understand the mechanism of 1,4-benzoquinone-induced cytotoxicity in platelets, the roles of ATP and calcium in platelet toxicity and morphological changes were investigated. Using scanning electron microscopy, morphological changes including membrane blebbing were observed in rat platelets 5 min after exposure to 1,4-benzoquinone, which were significantly different from shape changes (pseudopod formation) observed in response to physiological agonists. Benzoquinone-induced membrane blebbing of platelets was associated with rapid depletion of intracellular ATP and was independent of the presence of extracellular calcium. Benzoquinone-induced platelet lysis observed between 20 and 30 min was dependent on extracellular calcium and associated with increased cytosolic calcium. Cytotoxicity induced by 1,4-benzoquinone was inhibited by antagonists of calmodulin, suggesting that calmodulin could play an important role in platelet toxicity. These results suggested that the progression of events for benzoquinone-induced cytotoxicity in platelets was as follows: 1,4 benzoquinone depletes intracellular ATP; membrane blebbing occurs; calcium homeostasis is disrupted, activation of calmodulin-dependent processes results; finally cytotoxicity occurs. PMID- 11853972 TI - Schizophrenia and schizotypal personality: a tribute to Peter H. Venables. AB - Peter Venables made multiple contributions to the field of schizophrenia and schizotypy, most notably in the areas of psychophysiology, neurocognition, and assessment. On the 50th anniversary of the start of his research career in 1951, a conference on schizophrenia and schizotypy was held in his honor in Tuscany, Italy. This special edition encapsulates many of the presentations given at that meeting, covering the areas of neurodevelopment, assessment, genetics, psychophysiology, neurocognition, brain imaging, psychopharmacology, intervention, and prevention. A key theme of this special edition concerns the integration of schizophrenia and schizotypy research in a manner that will allow for a more comprehensive understanding of both of these disorders. PMID- 11853971 TI - Single chain antibodies specific for fatty acids derived from a semi-synthetic phage display library. AB - The biological activities of many acylated molecules are lipid dependent. Lipids, however, are poorly immunogenic or non-immunogenic. We employed a phage display semi-synthetic human antibody library to isolate anti-lipid antibodies. Selection was done against methyl palmitate, a 16 carbon aliphatic chain, and a major component of bacterial glycolipids and lipoproteins in animal cells. The selected single chain variable fragment (scFv) bound specifically to a 16 carbon aliphatic chain and to a lesser extent to a 14 or 18 carbon aliphatic chain and poorly to either 12, 22 or 8 carbon aliphatic chains. Furthermore, the scFv prevented micelle formation of lipoteichoic acid from Gram-positive bacteria; inhibited lipopolysaccharide-induced tumor necrosis factor alpha release in mononuclear cells; bound to hydrophobic bacterial surfaces, especially those of Gram-positive bacteria, and bound to Lck, a mammalian palmitated lipoprotein. Our data suggest that the phage antibody library can be successfully employed to obtain human anti aliphatic scFv human antibody fragment with potential therapeutic applications in neutralizing the deleterious effects of bacterial toxins as well as in structure- function analysis of lipoproteins in animal cells. PMID- 11853973 TI - Adult schizotypal personality characteristics and prenatal influenza in a Finnish birth cohort. AB - Recent evidence suggests that schizophrenia may result from a disruption of normal brain development during a critical, prenatal risk period in the 6th month of gestation. The phenotypic diagnostic manifestation of a basic genetic neurodevelopmental disorder may consist of characteristics approximated by the DSM-IV diagnosis of "schizotypal personality disorder" (SPD). We identified male conscripts in Finland who, as fetuses, were exposed to the 1969 Hong Kong Influenza epidemic, along with a group of controls born during a relatively low year (1971) for infectious epidemics. It was hypothesized that among fetuses exposed to the influenza epidemic in their 6th month of gestation, we would observe an increased frequency of elevated (upper quartile) scores on a schizotypal personality characteristics (SPC) scale as compared to controls. A significantly higher proportion of the 6th month index exposed subjects (39%) had "elevated" SPC scale scores as compared to their controls (26%) (p<0.003). Further analyses revealed that these differences were accounted for by those exposed to the influenza epidemic in week 23 (51% vs. 24%) of the 6th month (p<0.005). Exploratory analyses for the other months did not reveal any significant differences. Implications and limitations of the week 23 findings are discussed. PMID- 11853974 TI - Pubertal neurodevelopment and the emergence of psychotic symptoms. AB - One of the chief epidemiological hallmarks of schizophrenia is its modal age at onset in early adulthood. Clinical onset is preceded by an adolescent period that is usually characterized by increasing adjustment problems. Recent theorizing about the etiology of schizophrenia has focused on postpubertal brain changes that may be involved in triggering the expression of vulnerability for schizophrenia. In this paper, we further examine the normal neurodevelopmental processes that occur in adolescence and the underlying role of hormonal factors in controlling the expression of genes that govern brain maturation. We then consider how postpubertal hormone changes might serve to trigger the expression of vulnerability genes that code for abnormal brain development. PMID- 11853975 TI - Input dysfunction, schizotypy, and genetic models of schizophrenia. AB - Peter Venables proposed that an input dysfunction, which causes the brain to lose its ability to control the flood of sensory information into its higher level processing areas, might be an important pathophysiological mechanism in schizophrenia. The hypothesis was part of his general belief that even the most severe psychopathology arises from aberrations in normal brain psychophysiology. Neurobiological and genetic investigations based on his initial observations include the demonstration that diminished inhibition of the auditory-evoked response to repeated stimuli is a genetically determined deficit, linked to one of the chromosomal loci that is also responsible for the part of the genetically transmitted risk for schizophrenia. Increasing evidence that schizophrenia is a multigenetic illness prompts reconsideration of the nature of schizotypy. Individual genes that convey part of the risk for schizophrenia may be quite common in the general population and cause relatively subtle changes in psychophysiology. Thus, as predicted by Venables, the substrates of schizotypy and schizophrenia may arise from variants in normal brain function. PMID- 11853976 TI - Schizotypal personality disorder inside and outside the schizophrenic spectrum. AB - The concept of schizotypal personality disorder has been heavily discussed since its introduction into the official classification of mental disorders in DSM-III. The aim of this study was to investigate the difference between schizotypal personality disorder within and outside the genetic spectrum of schizophrenia. Schizotypals with and without schizophrenic cotwins and first-degree relatives were compared, with individuals with other mental disorders and no mental disorders as controls. It appeared that only inadequate rapport and odd communication were more pronounced among schizotypals within, compared to schizotypals outside the schizophrenic spectrum. Schizotypals outside the schizophrenic spectrum, however, scored higher than schizotypals inside the schizophrenic spectrum on ideas of reference, suspiciousness, paranoia, social anxiety, self-damaging acts, chronic anger, free-floating anxiety and sensitivity to rejection. Interestingly, the four last features are seldom observed among schizotypals inside the schizophrenic spectrum. Monozygotic non-schizophrenic cotwins of schizophrenics score high on inadequate rapport, odd communication, social isolation and delusions/hallucinations. Monozygotic non-schizophrenic cotwins of schizotypals outside the schizophrenic genetic spectrum score high on illusions, depersonalization, derealization and magical thinking. Negative schizotypal features appear to be inside the schizophrenic spectrum, while positive borderline-like features are outside having another genetic endowment. PMID- 11853977 TI - Does the Schizotypal Personality Questionnaire reflect the biological-genetic vulnerability to schizophrenia? AB - We investigated whether the Schizotypal Personality Questionnaire (SPQ) [Schizophr. Bull. 17 (1991) 555.] could be an indicator of the biological-genetic vulnerability to schizophrenia. We hypothesized that the mean scores on three dimensions of the SPQ of different groups of relatives of patients with schizophrenia would parallel their risk for developing schizophrenia. The SPQ was administered to 51 first-episode schizophrenia patients, 63 parents of schizophrenia patients, 42 siblings of schizophrenia patients and 12 children of schizophrenia patients. Patients differed from the relatives on all three dimensions. Siblings and children scored significantly higher than parents on Positive Schizotypy, and the insignificant difference between the siblings and children was in the expected direction. The results could not be explained by the differences in age, sex, IQ or substance abuse. No differences were found for Disorganization Schizotypy between the relatives. Children scored higher than parents on Negative Schizotypy. The current study offers support to the hypothesis that the positive dimension of SPQ reflects the genetic vulnerability to schizophrenia. PMID- 11853978 TI - Endophenotyping schizotypy: a prelude to genetic studies within the schizophrenia spectrum. AB - Schizophrenia is a complex genetic disease with a prevalence rate of 1% in the general population. Schizotypal personality disorder (SPD) occurs in up to 3% of the population, and these subjects are phenomenologically and perhaps genotypically related to schizophrenia. The diagnosis of SPD was empirically derived based on the symptoms of individuals with a genetic relationship to schizophrenia patients and SPD may be a more common phenotypic expression of a schizophrenia-related diathesis than is schizophrenia itself. Family-genetic studies have determined that (1) relatives of schizophrenic patients have an increased risk of SPD and (2) relatives of SPD subjects have increased the rates of both schizophrenia and SPD. Because SPD subjects do not typically have the confounding effects of a chronic illness, long-term hospitalization or chronic neuroleptic treatment, they are ideal for the study of the proposed trait-related vulnerability markers in schizophrenia spectrum individuals. The study of vulnerability markers in SPD subjects has become increasingly important because it provides a means of assessing phenotypic traits that may not be evident clinically. By combining multiple inhibitory/gating information processing measures, it may be possible to identify a subgroup of SPD subjects with multiple inhibitory deficits who are phenotypically most similar to patients with schizophrenia. Composite phenotypes can also be developed, which increase the probability of identifying the complex genetic architecture of schizophrenia spectrum disorders, which interact with nongenetic protective and exacerbating factors. PMID- 11853979 TI - Psychotic symptoms in non-clinical populations and the continuum of psychosis. AB - A growing body of evidence suggests that delusional or hallucinatory experiences are much more frequent in subjects from the general population than the prevalence of cases of psychotic disorders, thereby suggesting the existence of a symptomatic continuum between subjects from the general population and clinical cases of psychosis. Exploring the risk factors modulating the expression of psychosis-like signs in non-clinical populations may better contribute to elucidate the etiology of psychosis than research restricted to subjects at the endpoint of the distribution of the psychotic dimension. The aim of this paper is to briefly review research investigating the distribution of psychotic symptoms in non-clinical populations, the developmental aspects of psychosis proneness, and the outcome characteristics of psychosis-prone subjects. PMID- 11853980 TI - Schizotypal dimensions in normals and schizophrenic patients: a comparison with other clinical samples. AB - Schizotypy is usually referred to as a "liability" to schizophrenia, but it could also be more generally referred to as nonspecific "psychosis-proneness". This study examined the structure of schizotypy in a normal sample and in a sample of schizophrenics through the administration of the Schizotypal Personality Questionnaire (SPQ). Within a "fully dimensional" approach to psychiatric disorders, this study also compared SPQ scores in clinical samples of subjects with bipolar, unipolar and obsessive-compulsive (OCD) disorders. The results suggested that the disorganized three-factor model, Cognitive-Perceptual, Interpersonal Deficits and Disorganization, underlies individual differences across samples of normal and schizophrenic patients. Furthermore, the SPQ measures discriminated schizophrenia and bipolars from major depression but not from OCD. PMID- 11853981 TI - Increased psychophysiological arousal and orienting at ages 3 and 11 years in persistently schizotypal adults. AB - Abnormalities of psychophysiological arousal and orienting are thought to predispose to schizophrenia, but there have been no prior studies of early psychophysiological functioning in "persistent schizotypals" who remain stably schizotypal across time. This study assessed skin conductance (SC) arousal and orienting at ages 3 and 11 years, electroencephalography (EEG) at age 11 years, and behavior problems at age 17 years in 52 individuals who were stably schizotypal from ages 17 to 23 years, and 104 normal controls. Schizotypy was assessed at age 17 with the Schizophrenism scale, and at age 23 with the Schizotypal Personality Questionnaire (SPQ). Schizotypals had significantly increased SC arousal and increased SC amplitudes to neutral and aversive stimuli at both ages 3 and 11, and also reduced slow-wave left hemisphere EEG power at the beginning of the Continuous Performance Task (CPT) at age 11. Group differences in SC were greater for aversive than neutral stimuli. Schizotypals also had higher age 17 behavior ratings on psychotic behavior, anxiety withdrawal, and conduct disorder. Findings indicate that heightened SC arousal and orienting in early childhood is a significant risk factor for later schizotypal personality, support a left hemisphere overactivation hypothesis of schizotypy, and illustrate the potential value of a longitudinal definition of persistent schizotypy. PMID- 11853982 TI - What does electrodermal activity tell us about prognosis in the schizophrenia spectrum? AB - Various dysfunctions in electrodermal activity (EDA) have been found in schizophrenic patients. The present paper reviews evidence that the electrodermal dysfunctions may carry prognostic information regarding subsequent symptoms, as well as social and occupational outcome. Although the findings are not entirely consistent, heightened electrodermal activity as indicated by frequent orienting responses to innocuous stimuli, elevated skin conductance level (SCL), and frequent non-specific skin conductance responses (NS-SCR) is most often associated with poor symptomatic, social, and occupational outcome in schizophrenic patients. There have been no studies that have directly examined electrodermal prognostic indicators in schizotypal individuals. However, high risk studies suggest that heightened electrodermal activity may be prognostic of poor outcome in schizotypals as well. Thus, abnormally high electrodermal arousal and reactivity is predictive of poor outcome in at least some patients. The theoretical implications of these findings and directions for further research are briefly discussed. PMID- 11853983 TI - A Janusian perspective on the nature, development and structure of schizophrenia and schizotypy. AB - A personal review is presented of the functional basis of activation, withdrawal and unreality, individual differences in schizophrenia that Venables et al. pioneered. Activated and withdrawn syndromes were delineated from the totality of symptoms by classifying unmedicated patients on the basis of lateral asymmetries in electrodermal responses. A neuropsychological syndrome translation led to a syndrome hemispheric imbalance model supported by a literature review disclosing widespread cortical and infracortical involvement extending to motoneurone excitability, with validation from tests of learning, memory and evoked responses including the P300. It is contended that the centrality of arousal, the extensive substrate and the evidence of asymmetry modification with recovery and treatment all implicate specific and nonspecific thalamo-cortical systems whose uncoupling may lead to dysfunction of input, cognition and to unreality symptoms (found inconsistently related to asymmetry). The three syndromes have developmental associations including immune competence, ventricular changes and lateral asymmetry, putative regressive neuronal changes in connectivity and electrocortical measures of connectivity, as well as sensory gating and anomalies of P50 suppression and habituation. Replication of the syndromal structure in psychometric schizotypy indicates that syndrome expression is based on the premorbid personality, compatible with evidence of early determinants of the approach/withdrawal balance in social encounters. Functional considerations for the nature of schizophrenia support neurophysiological approaches to treatment such as neurofeedback. PMID- 11853984 TI - Eye movement and neuropsychological studies in first-degree relatives of schizophrenic patients. AB - The aim of the study was to compare the results of oculomotor and neuropsychological tests in first-episode schizophrenic patients, in both their parents and matched healthy controls. Eye movement tests included fixation and a smooth pursuit task and neuropsychological tests which comprised the Trail Making Test (TMT) A and B, the Stroop Test A and B, and the Wisconsin Card Sorting Test (WCST). There was a significant difference between the 21 patients and their 33 healthy parents in the results of both the eye movement tests and the TMT and WCST but not in the Stroop test. On the other hand, a significant difference between parents and their matched control subjects was found in both oculomotor tests, in the Stroop B, and in two indices of the WCST (completed categories, CC and percentage of conceptual level responses, %CONC). A correlation was obtained between patients and their fathers in the intensity of smooth pursuit disturbances and two indices of the WCST (CC and %CONC), but not between patients and their mothers. The results obtained confirm those of other studies pointing to the presence of a significant impairment on oculomotor and neuropsychological tests in first-degree nonpsychotic relatives (parents) of schizophrenic patients, which may be used as an endophenotypic markers of genetic predisposition to schizophrenia. PMID- 11853985 TI - Neurocognitive functioning and schizophrenia spectrum disorders can be independent expressions of familial liability for schizophrenia in community control children: the UCLA family study. AB - This study provided a further test of the hypothesis that certain neuromotor, language and verbal memory dysfunctions reflect genetic predisposition to schizophrenia, by examining the effects of family loading for schizophrenia (FLS) in normal controls without personal histories of schizophrenia or attention deficit hyperactivity disorder. In a case control design, 11 community controls (CC) with FLS were compared to 47 CC without FLS on tests of expressive and receptive language, visual motor coordination, full scale intelligence and verbal memory. In this study, FLS primarily reflects the incidence of schizophrenia spectrum diagnoses in the second-degree relatives of CC probands. CC probands with FLS had significantly poorer general intelligence, expressive and receptive vocabulary abilities, visual motor coordination and slower motor speed than CC probands without FLS. The variance in neurocognitive functioning associated with FLS is not due to the presence of any psychiatric disorders in CC probands, nor the presence of schizophrenia spectrum disorders in their parents. The relation between FLS and neurocognitive and neuromotor functioning in CC probands was moderated by the parent's cognitive functioning. The results of the present study indicate that familial liability to schizophrenia can be transmitted across two generations, independent of the presence of schizophrenia spectrum disorders in either the parent or proband, and account for significant variance in proband neurocognitive and neuromotor functioning. These findings suggest the neurocognitive and neuromotor functioning and schizophrenia spectrum disorders can be relatively independent expressions of familial liability to schizophrenia. PMID- 11853986 TI - The structure of schizotypy: relationships between neurocognitive and personality disorder features in relatives of schizophrenic patients in the UCLA Family Study. AB - Schizotypal personality features and certain neurocognitive deficits have been shown to aggregate in the relatives of schizophrenic patients, supporting the view that both are likely to reflect genetic contributions to liability to schizophrenia. Within the relatives of schizophrenic patients, however, the interrelationships between these potential indicators of liability to schizophrenia are not well known. Using data from the UCLA Family Study, we examine the interrelationships between personality disorder symptoms and neurocognitive functioning in nonpsychotic first-degree relatives of schizophrenic patients. Factor analyses indicate that several dimensions of schizotypy can be identified. A neurocognitive dysfunction dimension includes loadings from measures of sequential visual conceptual tracking, rapid perceptual encoding and search, and focused, sustained attention as well as the rating of odd and eccentric behavior from schizotypal personality disorder. Other aspects of schizotypal personality disorder form separate positive schizotypy and negative schizotypy dimensions. These analyses support the view that schizotypy is multidimensional in relatives of schizophrenic patients and indicate that neurocognitive deficits in perception and attention are associated with particular schizotypal personality features. PMID- 11853987 TI - Input dysfunction and beyond--an evaluation of CPT components. AB - The aim of our project is to analyze the functional meaning of neurocognitive components of the Continuous Performance Tests (CPT), which may be responsible for the well-documented performance deficit. Since the CPT can be considered as a vulnerability marker for schizophrenia, this question is of special interest. We set up a test battery testing five different cognitive processing modes: perceptual organization, selective attention, short-term memory (storing component), working memory (rehearsal component), and vigilance/sustained attention. In order to avoid the pitfall of interpreting results confounded by psychometric differences within tasks, we created psychometrically parallel versions within each experimental block (following the proposals of Chapman and Chapman [J. Nerv. Ment. Dis. 171 (1983) 658]). At the main experimental session, we tested newly admitted patients with a DSM diagnosis of schizophrenia during remission (N=30), patients with major depressive disorder (MD) (N=18), and healthy controls (N=20). Results showed that differences specific for schizophrenia are seen at the experimental block, which tests perceptual organization. However, all levels of perceptual organization performance were concerned, i.e., from processing organized to non-organized patterns. The regression analysis showed that 3-7 CPT version performances could be explained by problems with short-term memory, sustained attention, and perceptual organization. In light of these findings, we discussed whether etiology of schizophrenia could be conceptualized as a circumscribed neurocognitive deficit or a multifunctional, multilocal deficit. PMID- 11853988 TI - Differential metabolic rates in prefrontal and temporal Brodmann areas in schizophrenia and schizotypal personality disorder. AB - In an exploration of the schizophrenia spectrum, we compared cortical metabolic rates in unmedicated patients with schizophrenia and schizotypal personality disorder (SPD) with findings in age- and sex-matched normal volunteers. Coregistered magnetic resonance imaging (MRI) and positron emission tomography (PET) scans were obtained in 27 schizophrenic, 13 SPD, and 32 normal volunteers who performed a serial verbal learning test during tracer uptake. A template of Brodmann areas derived from a whole brain histological section atlas was used to analyze PET findings. Significantly lower metabolic rates were found in prefrontal areas 44-46 in schizophrenic patients than in normal volunteers. SPD patients did not differ from normal volunteers in most lateral frontal regions, but they had values intermediate between those of normal volunteers and schizophrenic patients in lateral temporal regions. SPD patients showed higher than normal metabolic rates in both medial frontal and medial temporal areas. Metabolic rates in Brodmann area 10 were distinctly higher in SPD patients than in either normal volunteers or schizophrenic patients. PMID- 11853989 TI - Elucidating continuities and discontinuities between schizotypy and schizophrenia in the nervous system. AB - Genetic epidemiological studies indicate that schizophrenia is a complexly inherited disorder, whereby multiple genes, additively and in interaction with environmental factors, contribute to increasing risk for phenotypic expression. Linkage studies have identified a number of chromosomal regions as likely to harbor susceptibility genes for schizophrenia, but the linked regions span relatively large chromosomal segments in each case. These efforts have been hindered in part by a lack of means to detect nonpenetrant carriers of predisposing genes and in part by uncertainties concerning the nature of the nongenetic factors involved and their mechanisms of action in relation to genetic factors. Our strategy has focused on elucidating changes in central nervous system structure, physiology, and function that mark an inherited genotype for schizophrenia and on identifying specific environmental and etiological contributors and their modes of action in relation to genetic factors. The research reviewed in this paper, making use of family and twin designs, suggests that certain neural system deficits in schizophrenia (e.g., prefrontal cortex, working memory) are reflective of an inherited diathesis to the disorder, while others (e.g., temporal cortex, episodic memory) result from the interacting influences of genetic factors and particular types of biologically disruptive environmental events (e.g., fetal hypoxia). These findings help to reveal continuities and discontinuities in the neural disturbances characteristic of the schizophrenia genotype and phenotype and encourage the use of quantitative neural trait measures in the search for schizophrenia susceptibility genes. PMID- 11853990 TI - Cognitive and brain function in schizotypal personality disorder. AB - Schizotypal personality disorder, a diagnosis defined partially in terms of a genetic relatedness to schizophrenia, has begun to receive extensive investigative study. While the exact etiologic relationship between schizotypal personality disorder and schizophrenia remains to be determined, three models have been considered: (1) the two may be distinct disorders, (2) they may be essentially identical disorders but expressed with different degrees of severity, or (3) they may be related disorders with a partially overlapping etiology that might account for the many similarities yet the lack of psychosis or severe deficits in schizotypal individuals. Some of the recent research in the structural and functional neuroanatomy, neurochemistry, cognitive function, and pharmacology of schizotypal personality disorder is reviewed with citation of the most recent findings from our laboratory and others. Both schizotypal and schizophrenic subjects appear to show abnormalities in temporal lobe volume, but schizotypal subjects do not appear to show the volumetric decreases in frontal cortex that schizophrenic patients evidence. Abnormalities in thalamic nuclei parallel these findings-the pulvinar, which projects to temporal association and sensory cortices, is reduced in both disorders, but the mediodorsal nucleus, which projects extensively to the frontal cortex, is reduced in schizophrenic patients but not in schizotypal patients. Functional imaging studies suggest that there may be abnormalities in frontal activation in both disorders, but that schizotypal individuals can recruit alternative regions to accomplish tasks requiring frontal lobe activation that may help compensate. Imaging studies of the subcortex including FDG/PET imaging of metabolic activity during a verbal learning task, SPECT imaging studies which measure binding of IBZM and its displacement following amphetamine administration, and plasma HVA determinations following 2-deoxyglucose administration all suggest the possibility of relatively reduced dopaminergic subcortical activity in schizotypal individuals compared to schizophrenic patients. Cognitive function is also impaired in the areas of working memory, verbal learning, and attention in schizotypal patients, as in schizophrenic patients, and they may be particularly susceptible to cognitive tasks with high context dependence, as in schizophrenia. Preliminary trials of catecholaminergic agents suggest that these agents may be able to improve these impaired cognitive functions. PMID- 11853991 TI - An integration of schizophrenia with schizotypy: identification of schizotaxia and implications for research on treatment and prevention. AB - The liability to schizophrenia (schizotaxia) is associated with deficits in a variety of domains, including negative symptoms and neuropsychological deficits, even in the absence of psychosis or pre-psychotic prodromal symptoms. Conceptually, this view of schizotaxia is similar to negative schizotypy (i.e., schizotypal personality disorder minus the positive symptoms). It is broader than DSM-IV schizotypal personality disorder (SPD), however, in that more relatives of patients with schizophrenia show core symptoms of schizotaxia than meet the diagnostic criteria for SPD. Three lines of evidence support the validity of schizotaxia. First, evidence of concurrent validation was obtained by showing that schizotaxic subjects were more impaired than non-schizotaxic subjects on a variety of independent clinical scales. Second, schizotaxic subjects showed higher levels of negative symptoms on the Structured Interview for Schizotypy than non-schizotaxic subjects, but did not differ on positive symptoms. Third, subjects who met predetermined criteria for schizotaxia (i.e., negative symptoms and neuropsychological deficits) showed positive effects following treatment with low doses of risperidone (0.25-2.0 mg). Thus, clinical deficits in schizotaxia may be identifiable, and to a significant extent, reversible. Implications for the conception of schizotypy and the prevention of schizophrenia will be discussed. PMID- 11853992 TI - The schizophrenia prodrome: treatment and high-risk perspectives. AB - Interest in the prodromal stage of schizophrenia-the stage directly preceding the onset of psychosis-has recently undergone a dramatic increase. To a great extent, this has resulted from the convergence of two very different research traditions. Many treatment researchers have moved from a concern with symptom control to an interest in prevention and view the prodrome as the optimal stage to begin intervention with anti-psychotics. High-risk researchers, who view the identification of accurate risk factors as necessarily preceding preventive programs, have begun to move from the premorbid to the prodromal phase as the most effective starting point. Thus, researchers in both traditions have targeted the schizophrenia prodrome as the most likely gateway to prevention. However, clashes between the two traditions in approaches, methodology and research goals have also led to considerable controversy. Such issues as how best to define the prodrome, what the actual risk for schizophrenia is among prodromal individuals, and what type of medication should be used remain largely unresolved. The Hillside Recognition and Prevention (RAP) Program has been designed to address many of these and related questions. Within a naturalistic treatment framework, the RAP program combines both high-risk and treatment research strategies. Preliminary findings from a 3-year RAP pilot study, for example, suggest that the prodrome is a developmentally complex phase of schizophrenia, that it consists of distinctly different subgroups and that novel anti-psychotics are clearly beneficial for some but not all individuals. Depending upon clinical characteristics and phase of the prodrome, anti-depressants also appear highly effective. PMID- 11853993 TI - Separase: a conserved protease separating more than just sisters. PMID- 11853994 TI - Actin' up (and down). AB - The Cote D'Azure provided a stunning backdrop for a recent conference* on the role of microfilament function and regulation in cell polarity. Although sadly missing several North American participants in the wake of events in Washington and New York, the largely European audience was nevertheless treated to fascinating insights into the many roles of actin polymerization and other processes involved in the establishment and maintenance of cell polarity. (*Joint CNRS/Jacques Monod/EMBO Conference on 'The Role of Microfilament Function and Regulation in Cell Polarity'; Presqu'ile de Giens, France 15-19 Sept 2001. Organized by Daniel Louvard and Tony Bretscher.) PMID- 11853996 TI - Polar wind left flapping in the breeze? PMID- 11853995 TI - Nups, Kaps, NXFs and others: making sense in the Babel of nuclear transport. AB - The spatio-temporal aspects of signal transduction, and the molecular basis of nucleocytoplasmic transport, have attracted much attention recently. How they might be linked was the theme of a recent conference*. Many aspects of nuclear transport were discussed and novel results presented, either in talks or posters, by the approximately 80 participants. (*EMBO workshop on 'Signal-transduction mediated regulation of nuclear transport'; 11-14 August 2001; Strasbourg, France. Organized by A.N. Malviya.) PMID- 11853997 TI - Specificity in signaling and modulators: the Homers. PMID- 11853998 TI - A tale of a traveling transcription factor. PMID- 11854004 TI - Pictures in cell biology. Actin-driven polar growth of plant cells. PMID- 11854005 TI - Direct modulation of the host cell cytoskeleton by Salmonella actin-binding proteins. AB - Invasive Salmonella trigger their own uptake into non-phagocytic eukaryotic cells by delivering virulence proteins that stimulate signaling pathways and remodel the actin cytoskeleton. It has recently emerged that Salmonella encodes two actin binding proteins, SipC and SipA, which together efficiently nucleate actin polymerization and stabilize the resulting supramolecular filament architecture. Therefore, Salmonella might directly initiate actin polymerization independently of the cellular Arp2/3 complex early in the cell entry process. This is an unprecedented example of a direct intervention strategy to facilitate entry of a pathogen into a target cell. Here, we discuss the Salmonella actin-binding proteins and how they might function in combination with entry effectors that stimulate Rho GTPases. We propose that membrane-targeted bacterial effector proteins might trigger actin polymerization through diverse mechanisms during cell entry by bacterial pathogens. PMID- 11854007 TI - Cdk5 behind the wheel: a role in trafficking and transport? AB - Cdk5, a serine/threonine kinase in the cyclin-dependent kinase (Cdk) family, is an important regulator of neuronal positioning during brain development. Cdk5 might also play a role in synaptogenesis and neurotransmission. Loss of Cdk5 in mice is perinatal lethal, and overactive Cdk5 induces apoptosis in cultured cells, indicating that strict regulation of kinase activity is crucial. Indeed, activity depends on the stability of activating partners, subcellular localization and the phosphorylation state of the enzyme itself. Deregulated kinase activity has been linked to neurodegenerative diseases such as Alzheimer's disease (AD) and amyotrophic lateral sclerosis (ALS). This review focuses on links between Cdk5 activity and components of cytoskeletal, membrane and adhesion systems that allow us to postulate a role for Cdk5 in directing intracellular traffic in neurons. PMID- 11854006 TI - Motor-cargo interactions: the key to transport specificity. AB - Eukaryotic cells organize their cytoplasm by moving different organelles and macromolecular complexes along microtubules and actin filaments. These movements are powered by numerous motor proteins that must recognize their respective cargoes in order to function. Recently, several proteins that interact with motors have been identified by yeast two-hybrid and biochemical analyses, and their roles in transport are now being elucidated. In several cases, analysis of the binding partners helped to identify new transport pathways, new types of cargo, and transport regulated at the level of motor-cargo binding. We discuss here how different motors of the kinesin, dynein and myosin families recognize their cargo and how motor-cargo interactions are regulated. PMID- 11854008 TI - Plakins: a family of versatile cytolinker proteins. AB - By connecting cytoskeletal elements to each other and to junctional complexes, the plakin family of cytolinkers plays a crucial role in orchestrating cellular development and maintaining tissue integrity. Plakins are built from combinations of interacting domains that bind to microfilaments, microtubules, intermediate filaments, cell-adhesion molecules and members of the armadillo family. Plakins are involved in both inherited and autoimmune diseases that affect the skin, neuronal tissue, and cardiac and skeletal muscle. Here, we describe the members of the plakin family and their interaction partners, and give examples of the cellular defects that result from their dysfunction. PMID- 11854009 TI - Cutting to the chase: calpain proteases in cell motility. AB - Calpains are a large family of intracellular proteases whose precise and limited cleavage of specific proteins might be an integral regulatory aspect of signaling pathways. This intriguing mechanism for transducing biochemical and biophysical information from the external milieu seems to operate during cell motility. The two first described and ubiquitous isoforms, mu-calpain and M-calpain, have been implicated in enabling cell spreading by modifying adhesion sites and in promoting locomotion of adherent cells by facilitating rear-end detachment. Recent elucidation of the molecular structure of calpain opens the door for understanding how these pluripotential signal proteins are regulated to help govern migration. Armed with this knowledge, the precise roles of calpains in inflammation, wound repair and tumor progression can be ascertained and offer novel therapeutic targets. PMID- 11854010 TI - Profile of Irving Weissman. PMID- 11854013 TI - Prolactin receptor diversity in humans: novel isoforms suggest general principles. PMID- 11854014 TI - Connecting islet development and developing useful islets. PMID- 11854015 TI - Protein kinase A and human disease. PMID- 11854019 TI - Nuclear receptor coregulators: multiple modes of modification. AB - Many proteins have been characterized as coregulators that can be recruited by DNA-binding nuclear receptors to influence transcriptional regulation. Recent genetic and biochemical studies have shown that cellular levels of coregulators are crucial for nuclear receptor-mediated transcription, and many coregulators have been shown to be targets for diverse intracellular signaling pathways and post-translational modifications. This review focuses on the different modes of regulation of nuclear receptor coregulators and the implications for tissue- and context-specific transcriptional responses to hormone and membrane receptor signaling. PMID- 11854020 TI - Aromatase inhibitors in breast cancer. AB - Several compounds that selectively inhibit estrogen synthesis via aromatase have been developed. Steroidal substrate analogs, such as formestane and exemestane, inactivate aromatase by binding irreversibly to it. Non-steroidal inhibitors, such as the triazole compounds letrozole and anastrozole, are highly potent, reversible inhibitors with good specificity for aromatase. The intratumoral aromatase model for postmenopausal breast cancer has been used to investigate the efficacy of letrozole, anastrozole and exemestane in combination and sequentially. Combining letrozole or arimidex with tamoxifen or faslodex was not more effective than the aromatase inhibitors alone, but was more effective than tamoxifen alone. Letrozole was superior to and longer lasting than the other agents, suggesting that aromatase inhibitors control tumor growth effectively by inducing greater tumor response and extending treatment time. In addition, aromatase inhibitors can be effective in patients relapsing from tamoxifen. Because two types of aromatase inhibitors are available, steroidal enzyme inactivators and reversible non-steroidal inhibitors in sequential therapy could be useful if resistance to one type develops. PMID- 11854021 TI - Diverse roles for the LDL receptor family. AB - The low-density lipoprotein (LDL) receptor family consists of several related scavenger receptors that not only function as important cargo transporters, but also inform the cell of changes in its environment by mediating signaling responses. The LDL receptor was the first family member to be characterized and its function seems to be restricted to lipoprotein metabolism. By contrast, lipoprotein metabolism does not appear to be the exclusive function of the other characterized LDL receptor family members. It is now apparent that cargo transport by members of the LDL receptor family is closely associated with regulation of cellular physiology and cellular signaling events. Here, we focus on the diverse biological activities of certain members of this family. PMID- 11854022 TI - Cell metabolism in the regulation of programmed cell death. AB - Increased cellular metabolism and resistance to apoptosis are two hallmarks of cell transformation. Recent progress in the understanding of the role of mitochondria in controlling apoptosis has brought attention to the links between elements of the apoptotic machinery and cellular metabolism. Here, we review the coordinated effects of growth factor withdrawal on bioenergetics and programmed cell death, and discuss the metabolic consequences of genes that prevent apoptosis, including the BCL2 family of genes and AKT. PMID- 11854023 TI - The marsupial model for male phenotypic development. AB - In all mammals, androgen formed in the developing testes is responsible for the aspects of male development in which the Wolffian ducts, urogenital sinus and urogenital tubercle are transformed into the epididymis/vas deferens, prostate and penis. That these events take place after birth in the marsupial makes it possible to examine male phenotypic development during pouch life. In the tammar wallaby, Macropus eugenii, the testicular androgen 5 alpha-androstane-3 alpha,17 beta-diol (5 alpha-adiol) is formed in the developing testis, is secreted into plasma and has the capacity to virilize female young pouch when administered exogenously. 5 alpha-Adiol is formed by immature testes in many species and appears to act in target tissues once it has been converted to dihydrotestosterone. PMID- 11854024 TI - ACRP30/adiponectin: an adipokine regulating glucose and lipid metabolism. AB - In recent years, we have learned that adipocytes are not merely inert storage depots for triglycerides but rather highly active cells with potent autocrine, paracrine and endocrine functions. Adipose tissue secretes a large number of physiologically active polypeptides. Although leptin remains one of the best studied examples of an adipocyte-specific secretory factor, recent reports describe potent physiological activities for another adipocyte-specific secreted protein, adipocyte complement-related protein of 30 kDa (Acrp30). Full-length versions of Acrp30 or its proteolytic fragments decrease the postprandial rise of plasma free fatty acids and improve postabsorptive insulin-mediated suppression of hepatic glucose output. A strong correlation between plasma Acrp30 levels and systemic insulin sensitivity is well established and the protein has putative anti-atherogenic properties that are relevant for the prevention of formation of atherosclerotic plaques. The current challenge is to understand the molecular mechanisms through which the protein exerts its multiple functions. PMID- 11854028 TI - CD163: a signal receptor scavenging haptoglobin-hemoglobin complexes from plasma. AB - CD163 is a highly expressed macrophage membrane protein belonging to the scavenger receptor cysteine rich (SRCR) domain family. The CD163 expression is induced by interleukin-6, interleukin-10 and glucocorticoids. Its function has remained unknown until recently when CD163 was identified as the endocytic receptor binding hemoglobin (Hb) in complex with the plasma protein haptoglobin (Hp). This specific receptor-ligand interaction leading to removal from plasma of the Hp-Hb complex-but not free Hp or Hb-now explains the depletion of circulating Hp in individuals with increased intravascular hemolysis. Besides having a detoxificating effect by removing Hb from plasma, the CD163-mediated endocytosis of the Hp-Hb complex may represent a major pathway for uptake of iron in the tissue macrophages. The novel functional linkage of CD163 and Hp, which both are induced during inflammation, also reveal some interesting perspectives relating to the suggested anti-inflammatory properties of the receptor and the Hp phenotypes. PMID- 11854029 TI - Cyclin E2, the cycle continues. AB - The eukaryotic cell cycle is regulated by a family of serine/threonine protein kinases known as cyclin-dependent kinases (CDKs). The activation of a CDK is dependent on its association with a cyclin regulatory subunit. The formation of distinct cyclin-CDK complexes controls the progression through the first gap phase (G(1)) and initiation of DNA synthesis (S phase). These complexes are in turn regulated by protein phosphorylation and cyclin-dependent kinase inhibitors (CKIs). Cyclin E2 has emerged as the second member of the E-type cyclin family. Cyclin E2-associated kinase activity is regulated in a cell cycle dependent manner with peak activity at the G(1) to S transition. Ectopic expression of cyclin E2 in human cells accelerates G(1), suggesting that cyclin E2 is rate limiting for G(1) progression. Although the pattern and level of cyclin E2 expression in some primary tumor and normal tissue RNAs are distinct from cyclin E1, both E-type cyclins appear to have inherent functional redundancies. This functional redundancy has facilitated the rapid characterization of cyclin E2 and uncovered unique features associated with each E-type cyclin. PMID- 11854030 TI - DNA polymerase beta. AB - Mammalian DNA polymerase beta(beta-pol) is a single polypeptide chain enzyme of 39kDa. beta-pol has enzymatic activities appropriate for roles in base excision repair and other DNA metabolism events involving gap-filling DNA synthesis. Many crystal structures of beta-pol complexed with dNTP and DNA substrates have been solved, and mouse fibroblast cell lines deleted in the beta-pol gene have been examined. These approaches have enhanced our understanding of structural and functional aspects of beta-pol's role in protecting genomic DNA. PMID- 11854031 TI - Cloning and characterization of ARHGAP12, a novel human rhoGAP gene. AB - Rho GTPase activating proteins (GAPs) stimulate the intrinsic GTP hydrolysis activity of Rho family proteins. Here we report the cloning of two splice variants of a novel gene named ARHGAP12 (access number), which has an ORF of 2541bp. Profilescan search result showed that its putative protein contains five domains: rhoGAP, SH3, PH and two WW domains. ARHGAP12 is located in chromosome 10pter-cen and consists of 20 exons according to the Blastn result against high throughput genomic sequences (htgs). Reverse transcription PCR and Northern blot indicates that it ubiquitously expresses in human tissues as well as tumor cell lines, suggesting its basic roles in cells. PMID- 11854033 TI - The purification and characterisation of m-calpain from ostrich brain. AB - Calpains are intracellular cysteine proteases activated in a Ca(2+)-dependent manner. The purpose of the present study was to investigate the physico-chemical and kinetic properties of ostrich brain m-calpain. m-Calpain was purified by successive chromatographic steps on Toyopearl-Super Q 650s and Pharmacia Mono Q HR 5/5 columns. A Ca(2+) concentration of 5mM and a casein concentration of 5mg/ml were found to be necessary for optimum calpain activity. Ostrich m-calpain exhibited a M(r) of 84K using SDS-PAGE and a M(min) of 79.3K from amino acid analysis. The pH and temperature optima were found to be 7.5 and 37 degrees C, respectively. The amino acid composition of m-calpain revealed 700 residues. The N-terminal sequence of m-calpain showed sequence identity with chicken (27%), human (23%) and rabbit (18%) and Schistoma mansoni (9%). PMID- 11854032 TI - Michaelis constants of mushroom tyrosinase with respect to oxygen in the presence of monophenols and diphenols. AB - The complex reaction mechanism of tyrosinase involves three enzymatic forms, two overlapping catalytic cycles and a dead-end complex. Analytical expressions for the catalytic and Michaelis constants of tyrosinase towards phenols and oxygen were derived for both, monophenolase and diphenolase activities of the enzyme. Thus, the Michaelis constants of tyrosinase towards the oxygen (K(mO(2))) are related with the respective catalytic constants for monphenols (k(M)(cat)) and o diphenols (k(D)(cat)), as well as with the rate constant, k(+8). We recently determined the experimental value of the rate constant for the binding of oxygen to deoxytyrosinase (k(+8)) by stopped-flow assays. In this paper, we calculate theoretical values of K(mO(2)) from the experimental values of catalytic constants and k(+8) towards several monophenols and o-diphenols. The reliability and the significance of the values of K(mO(2)) are discussed. PMID- 11854034 TI - Melatonin increases the activity of the oxidative phosphorylation enzymes and the production of ATP in rat brain and liver mitochondria. AB - We recently showed that melatonin counteracted mitochondrial oxidative stress and increased the activity of the mitochondrial oxidative phosphorylation (OXPHOS) enzymes both in vivo and in vitro. To further clarify these effects, we studied here the activity of OXPHOS enzymes and the synthesis of ATP in rat liver and brain mitochondria in vitro. In sub-mitochondrial particles, melatonin increases the activity of the complexes I and IV dose-dependently, the effect being significant between 1 and 10nM. Blue native-PAGE followed by histochemical analysis of the OXPHOS enzymes further showed the melatonin-induced increase of complex I activity. Titration studies show that melatonin counteracts the partial inhibition of complex IV induced by 5 microM potassium cyanide. However, melatonin (up to 5mM) was unable to recover the activity of complex IV when it was completely blocked by 100 microM cyanide. These data suggest that the indoleamine could stimulate the activity of the non-inhibited part of the complex IV. Melatonin also increases the production of ATP in control mitochondria and counteracts the cyanide-induced inhibition of ATP synthesis. These results provide new hormonal mechanism regulating mitochondrial homeostasis and may explain, at least in part, the anti-aging and neuroprotective properties of melatonin. PMID- 11854035 TI - Linear mixed irreversible inhibition of the autocatalytic activation of zymogens. Kinetic analysis checked by simulated progress curves. AB - Autocatalytic zymogen activation is a phenomenon of great importance for understanding some fundamental physiological processes involved in the enzyme regulation of gastrointestinal-tract enzymes, blood coagulation, fibrinolysis and the complement system. Examples of such processes are the activation of prekallikrein, trypsinogen and pepsinogen, all of which are controlled by natural proteinase inhibitors. This work studies the kinetics of a general autocatalytic zymogen activation process overlapped by two two-step irreversible inhibitions, i.e. a linear mixed irreversible inhibition. The kinetic equations for the whole course of the reaction are derived for this mechanism. In addition, we determine the corresponding kinetics for a number of particular cases of the general model analyzed, i.e. for reversible and irreversible non-competitive, competitive and uncompetitive inhibition systems which are considered particular cases of the general mechanism studied. The kinetic behavior of the system is related to a parameter, a dimensionless quantity, which shows whether the inhibition or the activation route prevails, in a similar way to that which we have previously carried out for other mechanisms. Finally, based on the kinetic equations obtained, a procedure for discriminating between the different mechanisms considered is suggested. The results of this contribution can be directly applied to most physiological autocatalytic zymogen activations in the presence of an inhibitor, allowing their complete kinetic characterization and suggesting procedures for varying the relative weight of the catalytic and inhibition routes or for changing the predominant route. PMID- 11854036 TI - Impaired capacity of acute-phase high density lipoprotein particles to deliver cholesteryl ester to the human HUH-7 hepatoma cell line. AB - The major role of native high density lipoprotein (HDL) is to carry cholesterol from peripheral tissues to the liver for bile excretion. As acute-phase (AP)-HDL has a decreased ability for cellular cholesterol efflux but an increased capacity for cholesteryl ester (CE) delivery to peripheral tissues, the interaction of AP HDL with human hepatoma cells was studied. Binding studies to HUH-7 cells revealed saturable binding properties for HDL and AP-HDL at 4 degrees C. At 37 degrees C, specific cell-association of (125)I- and [1,2,6,7-(3)H]-cholesteryl palmitate ([(3)H]CE)-labeled lipoprotein particles was 2.2- and 1.6-fold higher for HDL indicating that total CE delivery was significantly (P<0.05) higher for HDL in comparison to AP-HDL. In parallel, selective CE uptake (the difference between total lipid uptake and holoparticle uptake) from AP-HDL was decreased compared with HDL. The fact that the capacity for cellular cholesterol efflux from HUH-7 cells is slightly impaired by AP-HDL (compared with HDL) is of support that scavenger receptor class B, type I (SR-BI), the only receptor so far known to mediate bi-directional lipid flux, might be involved in altered HUH-7 cholesterol hemostasis by AP-HDL. Our in vitro findings suggest that HDL and AP HDL interact differently with cells of hepatic origin resulting in decreased hepatic cholesterol removal from the circulation during the AP reaction. PMID- 11854037 TI - IL-1beta activates C/EBP-beta and delta in human enterocytes through a mitogen activated protein kinase signaling pathway. AB - The enterocyte is an active participant in the inflammatory and metabolic response to sepsis, endotoxemia and other critical illnesses and is the site for cytokine and acute phase protein production in these conditions. The role of the CCAAT/enhancer binding protein (C/EBP) family of transcription factors in the response to inflammatory stimuli in the enterocyte is not well understood. In the present study, we treated Caco-2 cells with IL-1beta and determined C/EBP DNA binding activity by electrophoretic mobility shift assay. The involvement of the alpha, beta, and delta isoforms was determined by supershift analysis and Western blot analysis of proteins from the nuclear fraction. The role of the mitogen activated protein kinase (MAPK) signaling pathway was assessed by treating cells with the MAPK inhibitor PD-98059. Treatment of the Caco-2 cells with IL-1beta resulted in increased CCAAT/enhancer binding protein DNA binding activity. Supershift analysis and Western blotting indicated that this response to IL-1beta mainly reflected the delta isoform, and to a lesser degree the beta isoform. Treatment of the cells with PD-98059 inhibited the IL-1beta-induced increase in beta and delta activity. The results suggest that members of the CCAAT/enhancer binding protein family of transcription factors are activated in enterocytes during inflammatory conditions characterized by high levels of IL-1beta. PMID- 11854038 TI - Bioactivities of ricin retained and its immunoreactivity to anti-ricin polyclonal antibodies alleviated through pegylation. AB - Ricin has long been employed to construct immunotoxins, whose efficacy was often undermined by immunogenicity. Pegylation (modification of proteins with polyethylene glycol, PEG) was one of those recently developed approaches to circumvent immunogenicity of legions of drugs. Herein, pegylation of ricin was found to have barely changed the RNA N-glycosidase activity and protein synthesis inhibiting activity of ricin, but remarkably altered the cytotoxicity of ricin on hepatoma cell line (BEL7404) or the immunoreactivity with polyclonal anti-ricin antibodies. This result suggested that the attached PEG or monomethyloxyl polyethylene glycol (mPEG) groups did not hinder ricin from hydrolyzing ribosomal RNA, but indeed covered some areas on the surface of ricin molecule, including those involved in the interaction with cellular receptors and epitopes recognized by polyclonal antibodies. Pegylation, masking certain epitopes of ricin, might contribute to alleviate the immunogenicity of the toxin. Approach in this work, if applied to thereby constructed immunotoxins, would help improve the prospective efficacy of these toxins. PMID- 11854039 TI - Fusogenic activity of reconstituted newcastle disease virus envelopes: a role for the hemagglutinin-neuraminidase protein in the fusion process. AB - Enveloped viruses, such as newcastle disease virus (NDV), make their entry into the host cell by membrane fusion. In the case of NDV, the fusion step requires both transmembrane hemagglutinin-neuraminidase (HN) and fusion (F) viral envelope glycoproteins. The HN protein should show fusion promotion activity. To date, the nature of HN-F interactions is a controversial issue. In this work, we aim to clarify the role of the HN glycoprotein in the membrane fusion step. Four types of reconstituted detergent-free NDV envelopes were used, on differing in their envelope protein contents. Fusion of the different virosomes and erythrocyte ghosts was monitored using the octadecyl rhodamine B chloride assay. Only the reconstituted envelopes having the F protein, even in the absence of HN protein, displayed residual fusion activity. Treatment of such virosomes with denaturing agents affecting the F protein abolished fusion, indicating that the fusion detected was viral protein-dependent. Interestingly, the rate of fusion in the reconstituted systems was similar to that of intact viruses in the presence of the inhibitor of HN sialidase activity 2,3-dehydro-2-deoxy-N-acetylneuraminic acid. The results show that the residual fusion activity detected in the reconstituted systems was exclusively due to F protein activity, with no contribution from the fusion promotion activity of HN protein. PMID- 11854040 TI - Characterization of platelet-derived growth factor-C (PDGF-C): expression in normal and tumor cells, biological activity and chromosomal localization. AB - The predicted platelet-derived growth factor-C (PDGF-C) polypeptide contains an N terminal CUB-like domain and a C-terminal domain with homology to members of the PDGF/vascular endothelial growth factor (VEGF) family. PDGF-C mRNA is widely expressed in normal tissues and does not appear to be up-regulated in the tumor cell lines tested. The PDGF-C gene was mapped to human chromosome 4q31-32. PDGF-C protein and the CUB domain of PDGF-C expressed in Escherichia coli, were able to stimulate proliferation of human artery smooth muscle cells, but were inactive on umbilical vein endothelial cells, osteoblasts, fibroblasts, skeletal muscle cells (SkMC), bovine chondrocytes, and rat myocardium cells. Although the mitogenic activity of PDGF-C and the CUB domain was only observed at concentrations ranging from 1 to 10 microg/ml, substitution of Cys(124) by Ser or deletion of Cys(124) significantly reduced the mitogenic activity. Our data suggest a possible role of the CUB domain of PDGF-C in addition to its role in maintaining latency of the PDGF domain. PMID- 11854041 TI - New opportunities in toxicology in the post-genomic era. PMID- 11854042 TI - Drug-induced islet growth: a novel treatment for diabetes? PMID- 11854043 TI - AGENT makes cyanide a useful killer. PMID- 11854044 TI - A succulent cure to end obesity. PMID- 11854047 TI - Formulating fortunes -- the tale of a medicated lozenge. PMID- 11854048 TI - Is telomerase the cure-all and end-all? PMID- 11854049 TI - Cytokine inhibition: a new therapeutic avenue for skeletal diseases. PMID- 11854051 TI - Current themes in microarray experimental design and analysis. PMID- 11854052 TI - Chemical genomics: discovery of disease genes and drugs. PMID- 11854053 TI - Stem cells: hype or hope? AB - Stem cells undergo self-renewal and differentiate into multiple lineages of mature cells. The identification of stem cells in diverse adult tissues and the findings that human embryonic stem cells can be proliferated and differentiated has kindled the imagination of both scientists and the public regarding future stem cell technology. These cells could constitute an unlimited supply of diverse cell types that can be used for cell transplantation or drug discovery. The new options raise several fundamental ethical issues. This review gives an overview of the scientific basis underlying the hope generated by stem cell research and discusses current ethical and funding regulations. PMID- 11854054 TI - Stereoselective synthesis from a process research perspective. AB - The process chemists' primary responsibility is to develop efficient and reproducible syntheses of pharmaceutically active compounds. This task is complicated when dealing with chiral molecules that often must be made as single isomers according to regulatory guidelines. The presence of any isomeric impurity in the final product, even in small amounts, is usually not acceptable. This requirement necessitates an exquisite understanding of the methods employed in the construction of chiral drugs. However, the chemistry available for this purpose is sometimes limited and often requires a significant amount of effort and creativity to be made both functional and consistent. PMID- 11854055 TI - The evolving role of information technology in the drug discovery process. AB - Information technologies for chemical structure prediction, heterogeneous database access, pattern discovery, and systems and molecular modeling have evolved to become core components of the modern drug discovery process. As this evolution continues, the balance between in silico modeling and 'wet' chemistry will continue to shift and it might eventually be possible to step through the discovery pipeline without the aid of traditional laboratory techniques. Rapid advances in the industrialization of gene sequencing combined with databases of protein sequence and structure have created a target-rich but lead-poor environment. During the next decade, newer information technologies that facilitate the molecular modeling of drug-target interactions are likely to shift this balance towards molecular-based personalized medicine -- the ultimate goal of the drug discovery process. PMID- 11854058 TI - Premalignant lesions of the oral mucosa. A discussion about the place of oral intraepithelial neoplasia (OIN). AB - Oral precancerous lesions are traditionally classified as leukoplakia, erythroplakia, erythroleukoplakia, and distinguished from precancerous conditions. Major attention is focused on leukoplakia, and no distinction made whether dysplasia is or not present. Malignant transformation is a multistep process that should be approached also from the histological, and not merely from the clinical standpoint. Intraepithelial neoplasia, a notion created for the uterine cervix and already extended to other mucosae, should be adapted to the oral mucosa and used as diagnostic term. OIN (oral intraepithelial neoplasia) is not only a change in terminology, but also a progress in the unifying concept of precursors of squamous cell carcinoma, suppressing the useless discussion between severe dysplasia and carcinoma in situ. Furthermore, grading lesions as low or high grade OIN increases diagnostic consistency. OIN is suspected on three clinical patterns reflecting histological changes: mosaic, irregular keratosis, erythroplakia (or intermediate aspects), but dysplastic mucosa may also appear normal clinically. PMID- 11854059 TI - Visual inspection in oral cancer screening in Cuba: a case-control study. AB - A case-control study was conducted to evaluate the efficacy of an on-going oral cancer screening programme in Cuba in preventing advanced oral cancer in Cuba. The cases for the study consisted of 200 oral cancer patients with stage III and IV disease. Three apparently healthy subjects per case, matched for sex, age (plus or minus 5 years) and residing within a perimeter of 200 m of the house with the case, and willing to be interviewed, were recruited as the controls. Information on socio-economic indicators, smoking, drinking, diet and screening history in the form of visits to the dentist were collected by personal interview with the subjects. Odds ratio (OR), with 95% confidence intervals (CI), of being diagnosed with an advanced oral cancer, in relation to the screening experience at the date of diagnosis of case, 1, 2, 3, 6, and 12 months prior to the date of diagnosis of case were estimated by conditional logistic regression for matched data. The odds ratio of advanced oral cancer associated with screening in relation to screening experience 3 months prior to the diagnosis of the case was 0.67 (95% CI: 0.46-0.95). The odds ratio was 0.91 (95% CI: 0.60-1.37) for a single screening test and 0.41 (95% CI: 0.24-0.68) for two or more tests. The protection offered by screening persisted up to 3 years since the last test. The results of a descriptive evaluation of the programme also revealed limited evidence towards a shift from advanced to early stages after the introduction of the programme. Nonetheless, the results should be interpreted with caution in view of the several limitations in the study, particularly the fact that screening history was established indirectly by interviews and advanced oral cancers constituted the cases, rather than those who died from the disease. PMID- 11854060 TI - Nasopharyngeal cancer WHO type II-III: monoinstitutional retrospective analysis with standard and accelerated hyperfractionated radiation therapy. AB - The aim of this study is to assess the impact of prognostic factors in patients with locoregionally advanced nasopharyngeal cancer (NPC), WHO type II-III, treated with two different radiation therapy (RT) schedules: standard radiation therapy (SRT), and accelerated hyperfractionated radiation therapy (HART), with or without sequential chemotherapy. Between January 1986 and December 1999, 78 consecutive NPC patients were treated either with SRT (until August 1993) or with HART (from September 1993). Of the 78 patients, 60 were males and 18 females, the median age was 56 years (range 14-83). Nine patients had a non-keratinizing carcinoma (WHO type II) and 69 an undifferentiated carcinoma (WHO type III). Five year overall survival rate (OS) was 62%. Two months after RT, 73 patients were in complete remission. Disease-free survival (DFS) rates at 5 years were: 85% for the HART and 59% for the SRT group, respectively. A multivariate analysis, age (hazard ratio, HR=4.17 for > or = 60 vs. <50 years) and N-stage (HR=3.56 for N3a N3b vs. N0-N1) were significant for survival, whereas N-stage (HR=8.23 for N3a N3b vs. N0-N1) and RT schedule (HR=0.30 for HART vs. SRT) were significant for DFS. In our experience, HART achieved higher DFS rates than SRT; however, HART did not favourably affect OS. Toxicity was comparable in the two RT schedules. PMID- 11854061 TI - Continuous selective intraarterial chemotherapy in combination with irradiation for locally advanced cancer of the tongue and tongue base. AB - We retrospectively evaluated the results of the concurrent combination therapy of selective continuous intraarterial chemotherapy and radiotherapy in 39 patients with locally advanced cancer of the tongue and tongue base between September 1992 and January 2000. Thirty patients were fresh cases (stage II, 10 patients; stage III, 15; stage IV, five) and nine were recurrent cases. The primary lesion was present in the mobile tongue in 33 patients and the tongue base in six. External irradiation (median dose, 48.6 Gy) was performed in all patients, and interstitial brachytherapy using an Au grain or Cs needle (median dose, 50 Gy) in 21. In intraarterial chemotherapy, a catheter was selectively inserted into the lingual artery via the superficial temporal artery, and carboplatin (CBDCA) was continuously infused (median dose, 460 mg/m(2)) concurrently with radiotherapy. In 13 patients with cervical lymph node metastasis, two courses of systemic chemotherapy with 5-FU (700 mg/m(2) x 5 days) and cisplatin (40-50 mg/m(2)x2 days) or its analog was also performed. In 37 (94.9%) of the 39 patients in whom this combination therapy was completed, the response rate was 94.6%. The 3-year local control rate, progression-free survival rate, and overall survival rate by Kaplan-Meier's method were 79.2, 53.2, and 58.9%, respectively. This combination therapy was effective for locally advanced cancer of the tongue and tongue base without causing severe adverse side effects, and a local control rate comparable to that by surgery can be expected. PMID- 11854062 TI - The role of MDM2 in the proliferative activity of ameloblastoma. AB - Ameloblastoma is a unique tumor in the oral and maxillofacial region with various levels of proliferative activity in each type. p53 is most commonly found to be mutated in human cancer and sometimes is overexpressed also in other lesions, such as ameloblastoma. Murine Double Minute 2 (MDM2) is able to physically associate with the p53 tumor suppressor and therefore block the growth suppressive functions of p53. In the present study, immunohistochemistry, western blotting and enzyme-linked immunosorbent assay for p53 mutant selective test were done. MDM2 was overexpressed in ameloblastoma and the results showed different numbers of MDM2 labeling index based on both WHO classification and cytological pattern of outer layer cells. Basal ameloblastoma, which has a high proliferative activity, had the highest MDM2 labeling index. We suggest MDM2 protein caused the high proliferative activity of ameloblastoma, especially in basal cell ameloblastoma. PMID- 11854063 TI - The mRNA expression of placental glutathione S-transferase isoenzyme in hamster buccal-pouch carcinomas using reverse transcription-polymerase chain reaction. AB - Placental glutathione S-transferase (GST-P) may facilitate cell proliferation and inhibit apoptosis, hence allowing for the expansion of a population of initiated tumor cells. The enhanced expression of GST-P at the protein level has been reported previously in chemically induced oral carcinomas in hamster buccal-pouch mucosa but the expression of GST-P at the mRNA level has not yet been demonstrated. The purpose of the present study was to assess the GST-P mRNA expression in 7,12-dimethylbenz[a]anthracene (DMBA)-induced hamster buccal-pouch carcinomas using a reverse transcription-polymerase chain reaction (RT-PCR). Thirty-five outbred, young (6 weeks old), male, Syrian golden hamsters (Mesocricatus auratus) were randomly divided into one experimental group (15 animals), and two control groups (10 animals each). Bilateral pouches of a group of 15 animals of the experimental group were painted with a 0.5% DMBA solution three times a week for 12 weeks while each animal of one of the control groups was similarly treated with mineral oil. Another control group of 10 animals was untreated throughout the experiment. Areas of dysplasia and squamous-cell carcinomas with a 100% tumor incidence developed in all of the DMBA-treated buccal pouches. The mineral oil-treated and untreated pouches revealed no obvious changes. Placental glutathione S-transferase mRNA was demonstrated to be present amongst all the 12-week DMBA-treated hamster buccal-pouch mucosa animals, but not for the untreated animals or the animals for which the buccal pouch was treated with mineral oil. Multiple potential regulatory pathways including gene amplification, enhanced mRNA stability, chromosomal translocation/gene rearrangement, and hypomethylation of the promoter region can contribute to the overexpression of GST-P mRNA in DMBA-induced hamster buccal-pouch carcinomas. Further study is necessary to completely understand which candidate mechanism(s) will contribute principally to the increased GST-P mRNA expression in oral experimental carcinogenesis. PMID- 11854064 TI - Parotid Warthin's tumour Bristol Royal Infirmary (1985-1995): a study of histopathology in 33 cases. AB - This is a study of 33 (32 patients) confirmed Warthin's tumours (adenolymphomas) treated surgically at Bristol Royal Infirmary (1985--1995) focussing specifically upon 17 salient histopathological features together with capsular measurement by micrometry. Twenty-four out of 33 tumours had 'thin' capsules (< or = 200 microm), whereas 5/9 'thick' capsules were associated with gross tumour degeneration. The degenerative features including cystic change, squamous metaplasia, corpora amylacea like bodies, etc. were quantified. Tumours were classified into subtypes (typical, stroma poor, etc). These variations were not associated with age or sex. In 20/33 tumours a marginal sinus suggested a lymph node origin. Fine needle aspiration cytology (FNA) smears showing degeneration features are a diagnostic problem, but histopathological features are reflected in such smears and their recognition enhances diagnostic precision, enabling controlled surgical enucleation, the ideal operation, to be performed in most cases. In this study only 12 tumours were so treated; 11 others received parotidectomy. The theories of histogenesis are reviewed and discussed. PMID- 11854065 TI - Prognostic role of p27(Kip1) expression in oral squamous cell carcinoma in Taiwan. AB - The cyclin-dependent kinase inhibitor p27(Kip1) can inhibit the G1 to S transition of the cell cycle and is a putative tumor suppressor. Decreased expression of p27(Kip1) protein has been correlated with poor prognosis in a variety of human tumors. We examined the expression of p27(Kip1) in oral squamous cell carcinoma (SCC), epithelial dysplasia (ED) and normal oral mucosa (NOM) using antibodies to p27(Kip1). Positive p27(Kip1) nuclear staining was detected in all the specimems from ED and NOM, whereas positive p27(Kip1) staining was observed in 16 of the 63 (25%) cases of oral SCC. The labeling index for p27(Kip1) protein was significantly reduced from NOM through ED to oral SCCs, indicating that changes of p27(Kip1) protein expression may be an early event in oral carcinogenesis in Taiwan. The Kaplan-Meier analysis showed patients with p27(Kip1)-positive tumors had significantly higher overall survival than those with p27(Kip1)-negative tumors in a total of 63 patients (P=0.015) and 47 patients with areca quid chewing habit (P=0.026). Multivariate analysis showed decreased p27(Kip1) protein expression was an independent significant predictor of poor overall survival in the patients with oral SCCs. These results indicate that p27(Kip1) protein expression may serve as a putative new adjuvant prognostic marker for routine assessment of oral SCC patients. PMID- 11854066 TI - Aberrations in the MTS1 tumor suppressor locus in oral squamous cell carcinoma lines preferentially affect the INK4A gene and result in increased cdk6 activity. AB - The signal transduction pathway regulated by the retinoblastoma tumor suppressor protein, pRB, is abrogated in the majority of human cancers. Using a series of cell lines derived from oral squamous cell carcinomas (SCCs) that were not subjected to radiation or chemotherapy treatment, we detected specific hyperactivity of cyclin dependent kinase (cdk) 6 but not cdk4. Subcellular localization studies showed a predominant nuclear localization of cdk6, demonstrating that this kinase was biologically active. The molecular basis for this aberration are mutations in the MTS1 locus of chromosome 9p21. This locus encodes two partially overlapping genes, the cdk inhibitor p16(ink4a), and p14(ARF), an inhibitor of mdm2-mediated degradation of p53. Our analysis demonstrates that the mutations of the MTS1 locus in oral SCC specifically target expression of the p16(ink4a) gene but less frequently affect p14(ARF). These results suggest that hyperactivity of cdk6 represents a distinct mechanism for pRB inactivation in oral SCC. PMID- 11854067 TI - A comparison of three comorbidity indexes in a head and neck cancer population. AB - We explored differences in prognostic ability for mortality of the established and validated Charlson comorbidity index with two other comorbidity indexes developed for this study. Our study was limited to persons diagnosed with HNCA between 1985 and 1993 in a database formed by a linkage of files from the National Cancer Institute's Surveillance, Epidemiology, and End Results Program with Health Care Finance Administration Medicare files (n=9386). Adjusted relative risks (RR) and 95% confidence intervals (95%CI) for comorbidity index scores of 1 or more compared to 0 were (RR=1.50, 95% CI 1.43-1.68) Charlson index, (RR=1.53 95% CI 1.42-1.66) HNCA index, and (RR=1.49, 95% CI 1.32-1.68) ATC index, respectively. The Charlson and HNCA indexes displayed dose-response patterns (P-value for trend <0.0001). Although the ATC index appears promising, the HNCA and Charlson indexes had similar adjusted RR's, dose-response patterns, P-values, and chi-square scores and appear particularly well-suited to the measurement of comorbidity. PMID- 11854068 TI - Increased tissue inhibitor of metalloproteinase-1 expression and inhibition of gelatinase A activity in buccal mucosal fibroblasts by arecoline as possible mechanisms for oral submucous fibrosis. AB - Oral submucous fibrosis (OSF) is a pre-malignant fibrotic lesion of the mouth in areca quid chewers. It is probably a consequence of disturbances in the hemeostatic equilibrium between synthesis and degradation of extracellular matrix molecules (ECM). To date, there has been little research about the role of tissue inhibitors of metalloproteinase (TIMPs) and matrix metalloproteinases (MMPs) in the pathogenesis of OSF. In the present study, we examined the activity of TIMPs from cells cultured from OSF and normal buccal mucosa. OSF specimens were found to have higher TIMP-1 expression than normal buccal mucosal fibroblasts (BMFs) by Western blots. To verify whether arecoline, a major areca nut alkaloid, could affect TIMP or MMP production by human BMFs, Western blots and gelatine zymography were used. Arecoline was found to elevate TIMP-1 expression at the concentration level under 20 microg/ml in a dose-dependent manner. The amount of TIMP-1 was about 2.7 fold at a concentration level of 10 microg/ml compared with control. From gelatin zymograms, the main gelatinolytic proteinase secreted by the human BMFs was MMP-2, and only minimal amounts of MMP-9 could be detectable from zymogram. In addition, arecoline was found to inhibit MMP-2 secretion and production at the concentration level of 40 microg/ml. The gelatinolytic activity of MMP-2 was about 54% at a concentration level of 80 microg/ml compared with control. Taken together, it was found that arecoline acted not only as an inhibitor on gelatinolytic activity of MMP-2, but also a stimulator for TIMP-1 activity. These synergistic effects may contribute to the ECM components accumulation in the areca quid associated OSF. PMID- 11854069 TI - Oral leiomyosarcoma: review of the literature and report of two cases with assessment of the prognostic and diagnostic significance of immunohistochemical and molecular markers. AB - Leiomyosarcoma of the oral cavity is a very rare tumor that is associated with aggressive clinical behavior and low survival. In this paper, we report two new cases of leiomyosarcoma affecting the mandibular gingiva and mandible of a 35 year-old male and the mandible of a 51-year-old female. Given the difficulty in the histopathologic discrimination between benign and malignant smooth muscle tumors and the absence of reliable histologic parameters for prognostication of leiomyosarcomas, we evaluated the diagnostic and prognostic value of various immunohistochemical and molecular markers. By means of immunohistochemistry and quantitative real-time PCR analysis, we detected protein expression of PCNA, bcl 2, CDK4, p53 and MDM2 in both our cases and MDM2 amplification in our second case. The literature, pertinent to oral leiomyosarcoma and to molecular analysis of smooth muscle tumors, is reviewed. PMID- 11854070 TI - Clear cell odontogenic carcinoma. AB - Clear cell tumours, in the head and neck region, are usually derived from salivary or odontogenic tissues, or may be metastatic. A few clear cells may be present in odontogenic cysts, while, odontogenic neoplasms composed predominantly of clear cells are quite rare. They include calcifying epithelial odontogenic tumours (CEOT), ameloblastoma and odontogenic carcinoma. Clear cell odontogenic tumour (CCOT) has been classified in the last WHO classification as a benign tumour, but it is now recognized as a more sinister lesion and current opinion is that CCOT should be designated as a carcinoma. These tumours are characterized by aggressive growth, recurrences, and metastatic disease. A recent review of the literature has yielded 30 cases of tumours with similar characteristics. These tumours have a peak incidence in the 5th-7th decades, with a female predilection. The anterior portions of the jaws, especially the mandible, are most frequently affected. The aggressive potential of these neoplasms is well documented by the extensive invasion of adjacent tissues, multiple recurrences and regional or distant metastases. PMID- 11854071 TI - Dysplastic enamel in peripheral ameloblastoma. PMID- 11854072 TI - Comment on: S.S. Prime et al. "A review of inherited cancer syndromes and their relevance to oral squamous cell carcinoma" Oral Oncology 2001;37(1), 1-16. PMID- 11854075 TI - Treatment of intestinal parasitic infections: a review of nitazoxanide. PMID- 11854076 TI - Cryptosporidium: from molecules to disease. AB - The international conference, Cryptosporidium: from molecules to disease, was held 7-12 October 2001, in Fremantle, Western Australia, to discuss all aspects of Cryptosporidium and cryptosporidiosis. PMID- 11854077 TI - Rodent models of malaria in the genomics era. AB - The Rodent Malaria Genomics Symposium: Current Status and Future Directions was held on 15-16 November 2001 in Atlanta, GA, USA. PMID- 11854079 TI - A double-headed pro-drug that overcomes chloroquine resistance. PMID- 11854080 TI - The benefits of multiple Plasmodium infections. PMID- 11854081 TI - Are the costs correct? PMID- 11854082 TI - Unexpected consequences of vaccination against parasites. PMID- 11854083 TI - Haemoglobin C for supermen and women. PMID- 11854085 TI - Current status of malaria rapid diagnostic devices: an update. PMID- 11854086 TI - Systematics of the parasitic Protozoa. PMID- 11854087 TI - The PacELF programme: will mass drug administration be enough? AB - The Pacific Programme to Eliminate Lymphatic Filariasis is a regional, mass drug administration-based campaign in 22 countries and territories with the aim of eliminating filariasis transmission and alleviating the suffering caused by Wuchereria bancrofti. The challenges to filariasis elimination campaigns based on mass drug-administration alone are reviewed in this article. These challenges together with the previous successes of mosquito control campaigns in eliminating filariasis from regions in the Pacific argue for inclusion of entomology components in the control of filariasis and the monitoring of filariasis elimination programs. PMID- 11854088 TI - Why should parasite resistance be costly? AB - Parasite resistance is sometimes associated with fitness costs. Costs of resistance are fundamentally important in epidemiology, and in the ecology and evolution of host-parasite interactions. The cost of resistance is often envisioned as the cost of re-allocating limiting resources to resistance machinery from other traits. This popular paradigm has resulted in a spate of research that assumes a fitness cost to resistance. We comment on this trend and propose a working framework of various resistance means and mechanisms. Within these means and mechanisms, we suggest that many are not likely to incur significant fitness costs. PMID- 11854089 TI - When is a parasite species a species? AB - Regrettably, 140 years after the publication of Darwin's Origin of Species, we face the grotesque situation that we still do not know what is a species whose origin Darwin wanted to explain. A generally applicable species definition is not available. Is there a basic unit of biodiversity above the level of individuals? Do we try to define something that does not exist in reality? The strong potential for the evolution of genetic variability in parasites together with the importance of species diagnosis for applied fields of parasite research make biodiversity research a key role in parasitology. Frequent occurrence of sympatric speciation, clonal reproduction, selfing, sib mating or parthenogenesis imply exceptional conditions for the evolution of gene pool diversities in parasites. PMID- 11854090 TI - Interpreting low praziquantel cure rates of Schistosoma mansoni infections in Senegal. AB - Praziquantel is currently the drug of choice for the treatment of schistosomiasis. Selective treatment of Schistosoma mansoni infections in various endemic countries usually present cure rates of >70% when using the standard dose of 40 mg kg(-1) body weight of praziquantel. However, unusually low cure rates (18-38%) have been reported from Senegal, raising fears for emergence of resistance (or tolerance) to praziquantel. One major problem is the precise quantitative interpretation of cure rates, which allows an unequivocal distinction between drug failure and normal drug performance. This article reviews studies on praziquantel treatment of population by standardizing the data through an innovative meta-analysis and provides empirical evidence concerning the extent to which the reported low cure rates from Senegal are atypical. PMID- 11854091 TI - A multilateral effort to develop DNA vaccines against falciparum malaria. AB - Scientists from several organizations worldwide are working together to develop a multistage, multigene DNA-based vaccine against Plasmodium falciparum malaria. This collaborative vaccine development effort is named Multi-Stage DNA-based Malaria Vaccine Operation. An advisory board of international experts in vaccinology, malariology and field trials provides the scientific oversight to support the operation. This article discusses the rationale for the approach, underlying concepts and the pre-clinical development process, and provides a brief outline of the plans for the clinical testing of a multistage, multiantigen malaria vaccine based on DNA plasmid immunization technology. PMID- 11854092 TI - Towards a recombinant antigen vaccine against Onchocerca volvulus. AB - Various approaches to identify potential vaccine candidates against onchocerciasis resulted in the cloning of recombinant proteins, which confer protection in vaccinated mice. The development of an effective vaccine against onchocerciasis has been the focus of a research program supported by the Edna McConnell Clark Foundation from 1985 to 1999. The approaches used to clone potential protective antigens and the successful vaccination of animals with some of the antigens are summarized here. PMID- 11854095 TI - Susceptibility gene discovery for common metabolic and endocrine traits. AB - Almost all major causes of ill-health and premature death in human societies worldwide - including cancer, cardiovascular disease, diabetes and many infectious diseases - are, at least in part, genetically determined. Typically, risk of succumbing to one of these illnesses is thought to depend on both the individual repertoire of variation within a number of key susceptibility genes and the history of exposure to relevant environmental factors. For many of these conditions, the molecular basis of disease pathogenesis remains obscure. This represents a major obstacle to development of improved, rational strategies for disease treatment, prevention and eradication. It is easy therefore to appreciate the importance attached to efforts to deliver more comprehensive understanding of the molecular basis of disease pathogenesis. Nor is it hard to understand that identification of major susceptibility genes should highlight those components of molecular machinery that are critical for the preservation of normal health. The benefits promised are great, but progress to gene identification in multifactorial traits has been rather disappointing to date. Why is this? This review aims to answer this question by describing current and future approaches to gene discovery in multifactorial traits. The examples quoted will mostly relate to type 2 diabetes, but the issues and approaches are generic, and apply equally to other multifactorial traits in the endocrine and metabolic arena - type 1 diabetes; obesity; hyperlipidaemia; autoimmune thyroid disease; polycystic ovarian syndrome - and beyond. PMID- 11854096 TI - Accumulation of rat pineal serotonin N-acetyltransferase mRNA induced by pituitary adenylate cyclase activating polypeptide and vasoactive intestinal peptide in vitro. AB - In mammals, pineal melatonin secretion is under the control of adrenergic and peptidergic inputs regulating serotonin N-acetyltransferase (arylalkylamine N acetyltransferase; AA-NAT) activity. In this study, the accumulation of AA-NAT mRNA induced by norepinephrine (NE) and peptides of the secretin superfamily (pituitary adenylate cyclase activating polypeptide (PACAP), vasoactive intestinal peptide (VIP), growth hormone releasing factor (GRF), secretin) was investigated by a new quantitative reverse transcription-PCR (RT-PCR) assay. We demonstrated that PACAP was the most potent peptide to increase the expression of AA-NAT mRNA and to induce cAMP production in rat pinealocytes. VIP was also able to elevate the AA-NAT mRNA level and cAMP efflux in a dose-dependent manner; however, it was six- and threefold, respectively, less potent than PACAP. The maximal values of AA-NAT mRNA level after PACAP and VIP exposures were similar (523.1 +/- 52.5 amol to 640.7 +/- 68.8 amol vs 461.5 +/- 54.3 amol to 579.2 +/- 72.4 amol). These saturable peak values were approximately five- to eightfold less than that after NE (3.0 +/- 0.3 fmol to 3.6 +/- 0.4fmol). GRF and secretin were less potent than VIP in inducing AA-NAT gene expression and cAMP efflux. These data suggest that the peptides act mostly on VIP(1)/PACAP (VPAC(1)) receptors of pinealocytes with different affinity. The peak cAMP efflux always preceded the elevation of AA-NAT gene expression during the 3-h infusion of VIP or NE. The cAMP efflux had declined by the time of onset of maximal AA-NAT gene expression, but remained significantly higher than its basal values. Our data indicate that even a submaximal level of cAMP is sufficient for maintaining the maximal AA-NAT mRNA accumulation. These findings show that, in addition to NE, PACAP and VIP may have an important role in the regulation of AA-NAT mRNA levels in rat pinealocytes. PMID- 11854097 TI - Expression profile of active genes in the human pituitary gland. AB - To characterize transcripts abundantly expressed in the human pituitary gland in general as well as to isolate novel transcripts expressed specifically in this gland, we generated an expression profile of the active genes transcribed in it. A total of 1015 randomly collected 3prime prime or minute expressed sequence tags (ESTs) (gene signatures, GSs) were grouped into 527GS species. The results showed the relative expression levels of genes in the pituitary gland. The genes comprising more than 1% of total mRNA were prolactin, growth hormone and chromogranin B genes. When known genes were categorized, the genes for pituitary hormones were the most actively transcribed, followed by the genes for ribosomal proteins, nuclear proteins and secretory granule proteins. Through comparison of this gene expression profile with the BodyMap database containing profiles generated from 63 other human tissues, we obtained 11 genes which appeared to be specifically expressed in the pituitary gland. In addition to the eight known genes, we identified three novel pituitary-specific transcripts which encode putative proteins: pituitary gland specific factor 1a (PGSF1a), PGSF1b and PGSF2. This expression profile method is a novel approach to the isolation of pituitary specific genes that may have important functions. PMID- 11854098 TI - Prolactin activation of mammary nitric oxide synthase: molecular mechanisms. AB - Prolactin (PRL) is capable of stimulating both calcium and nitric oxide (NO) accumulation in mammary epithelial cells within 15min. A calcium ionophore was also able to stimulate NO levels to an extent similar to that generated by PRL. Furthermore, maximal concentrations of PRL and the ionophore were not additive, suggesting that they were both using the same pathway, i.e. calcium. Finally, the depletion of intracellular calcium completely abrogated the effect of PRL on NO production. No other pathway known to affect NO synthase (NOS) influenced the action of PRL. Specifically, manipulations of protein phosphatase 2B, protein kinase B (PKB), protein kinase C (PKC), and arginine transport did not alter the activation of NOS by PRL. Therefore, the ability of PRL to stimulate NO production at 15min can be completely explained by its ability to elevate intracellular calcium. PMID- 11854099 TI - Antagonistic effects of 24R,25-dihydroxyvitamin D3 and 25-hydroxyvitamin D3 on L type Ca2+ channels and Na+/Ca2+ exchange in enterocytes from Atlantic cod (Gadus morhua). AB - There is mounting evidence that vitamin D and its metabolites play important roles in regulating plasma calcium concentrations in teleost fish as in other vertebrates. The aims of the present study were to elucidate the possible cellular target mechanisms for the rapid actions of 24R,25(OH)(2)D(3), 25(OH)D(3) and 1,25(OH)(2)D(3) in Atlantic cod enterocytes at physiological doses, and to establish the concentration and thus the physiological range of circulating 24R,25(OH)(2)D(3), 25(OH)D(3) and 1,25(OH)(2)D(3) in the Atlantic cod. The plasma concentrations of 25(OH)D(3), 1,25(OH)(2)D(3) and 24R,25(OH)(2)D(3) were 15.3 +/- 2.7nM, 125.1 +/- 12.3pM and 10.1 +/- 23.5nM respectively. Exposure of enterocytes to 10mM calcium (Ca(2+)) evoked an increase in intracellular Ca(2+) concentrations ([Ca(2+)](i)). This increase was suppressed by 24R,25(OH)(2)D(3) dose-dependently, with an EC(50) of 4.9nM and a maximal inhibition of 60%. 24R,25(OH)(2)D(3) (20nM) abolished an increase in [Ca(2+)](i) (approximately 252%) in the control enterocytes exposed to 10microM S(-)-BAYK-8644, suggesting that the hormone acts by inhibiting Ca(2+) entry through L-type voltage-gated Ca(2+) channels. Administration of 20nM 24R,25(OH)(2)D(3) to enterocytes in the absence of extracellular Ca(2+) increased [Ca(2+)](i) by approximately 20%, indicating a release of Ca(2+) from intracellular stores. Administration of 25(OH)D(3) (20nM) resulted in a biphasic change in the enterocyte [Ca(2+)](i): within 1--5s, it decreased to 87 +/- 12nM below its mean basal [Ca(2+)](i) (334 +/- 13nM), followed by a rapid recovery of [Ca(2+)](i) to a new level, 10% lower than the initial [Ca(2+)](i). The rapid decrease, the recovery rate and the final [Ca(2+)](i) were all affected dose-dependently by 25(OH)D(3), with EC(50) values of 8.5, 17.0 and 18.9nM respectively. Furthermore, the effects of 25(OH)D(3) were sensitive to sodium (Na(+)), bepridil (10microM) and nifedipine (5 microM), suggesting that 25(OH)D(3) regulates the activity of both basolateral membrane associated Na(+)/Ca(2+) exchangers and brush border membrane-associated L-type Ca(2+) channels. Administration of 25(OH)D(3) (10nM) to enterocytes in the absence of extracellular Ca(2+) increased [Ca(2+)](i) by approximately 18%, indicating a release of Ca(2+) from intracellular stores. 1,25(OH)(2)D(3) also affected enterocyte [Ca(2+)](i) in a biphasic manner: the rapid decrease, the recovery rate, and the mean final [Ca(2+)](i) were all affected dose-dependently, with EC(50) values of 8.3, 24.5 and 7.7nM respectively. The high EC(50) values for 1,25(OH)(2)D(3) compared with circulating concentrations of 1,25(OH)(2)D(3) (130pM) suggest that this effect is pharmacological, rather than of physiological relevance in enterocyte Ca(2+) homeostasis of the Atlantic cod. It is concluded that 24R,25(OH)(2)D(3) has a physiological role in decreasing intestinal Ca(2+) uptake via inactivation of L-type Ca(2+) channels, whereas the physiological role of 25(OH)D(3) is to increase enterocyte Ca(2+) transport via activation of Na(+)/Ca(2+) exchangers, concurrent with activation of L-type Ca(2+) channels. PMID- 11854100 TI - Palliative care in neurology. AB - This article defines the evolving role of the neurologist as a provider of palliative care. As neurologists care for chronically and critically ill, dying patients, and individuals whose diseases are incurable at the time of diagnosis, clinical competence requires expertise in the principles and practice of palliative medicine. Multiple studies suggest that despite available guidelines many patients with neurological disease suffer from pain, dyspnea, and other symptoms at or near the end of life. Recommendations from the American Academy of Neurology and Institute of Medicine are provided and the many ongoing educational efforts aimed at closing the existing gap in knowledge and improving patient care are reviewed. PMID- 11854101 TI - Palliative care in patients with multiple sclerosis. AB - Multiple Sclerosis (MS) is an inflammatory demyelinating disease of the central nervous system (CNS) and can be characterized by acute exacerbations or gradual worsening of neurological function and disability. The course of the disease is highly variable and unpredictable, however, there are short and long-term favorable and unfavorable predictive factors, which may provide some information about the future pattern of the disease. Palliative care in MS is directed at symptom management, psychosocial support, and rehabilitation. The goal in palliative care is to achieve a high quality of life. The disease modifying agents, interferon beta, Glatiramer acetate and Mitoxantrone are the mainstay of treatment in MS. Symptomatic relief and counseling of patients with MS have a strong impact on quality of life. PMID- 11854102 TI - Palliative care in amyotrophic lateral sclerosis. AB - Amyotrophic lateral sclerosis (ALS) is the most common degenerative motor neuron disease in adults. The clinical picture consists of generalized fasciculations, progressive atrophy and weakness of the skeletal muscles, spasticity and pyramidal tract signs, dysarthria, dysphagia, and dyspnea. Pseudobulbar affect is common. Disease-specific treatment options are still unsatisfactory. Therapeutic nihilism is not justified as a large array of palliative measures available to enhance the quality of life of patients and their families. Because of its clinical characteristics, ALS represents a paradigm for palliative care in neurological diseases. Numerous projects are being undertaken worldwide in an effort to enlarge the evidence base for palliative interventions in ALS. Palliative care in ALS is a multidisciplinary effort requiring careful coordination. An open and empathic disclosure of the diagnosis is essential. Nutritional deficiency caused by dysphagia can be relieved by a percutaneous endoscopic gastrostomy. Respiratory insufficiency can be effectively treated by non-invasive home mechanical ventilation. The terminal phase of the disease should be discussed at the latest when symptoms of dyspnea appear, in order to prevent unwarranted fears of "choking to death." Psychological and spiritual care of patients and families are important. Collaboration with hospice institutions and completion of advance directives can be of invaluable help in the terminal phase. PMID- 11854103 TI - Palliative care for people with late-stage Huntington's disease. AB - Huntington's disease provides a unique model to examine issues of long-term and palliative care for a younger and more mobile population who remain institutionalized longer than other nursing home residents. A lack of community based alternatives for young families, and the need for highly complex care planning encourage long term care placement mid-way in the disease process. Despite the need for a body of knowledge specific to environmental enrichment and therapeutic strategies for improving quality of life for people in the later stages of Huntington's disease, there is no published data in the neurological or rehabilitation literature. This article reviews the signs and symptoms and offers a multidisciplinary approach to ameliorate problems frequently encountered in caring for this population. PMID- 11854104 TI - Dementia. AB - Alzheimer's disease is the most common cause of dementia, but often several other conditions causing dementia are present on brain autopsies. Palliative care medical issues are similar in all late stage progressive degenerative dementias and include; cardiopulmonary resuscitation, transfer to acute care setting, antibiotic therapy, and tube feeding. Behavioral symptoms of dementia include agitation and resistiveness to care. Quality of life of individuals suffering from dementia is enhanced by availability of meaningful activities and by avoidance of restraints. Family support and involvement are crucial for optimal care. Formulation of advanced directives or an advance proxy plan is important for assuring the patient's previous wishes or best interests are considered when decisions about treatment strategies are made. PMID- 11854105 TI - Brain tumors. AB - The total care of a patient with an incurable brain tumor is a complex task. It is best performed by a team of specialized care providers including neurologists, neuro-oncologists, neurosurgeons, radiation oncologists, nurses, neuropsychologists, social workers, and physical and occupational therapists. Hospice providers, and spiritual counselors provide particular skilled services and comfort to patients nearing the end of life. The role and prominence of each team member evolves during the course of the illness as the disease progresses and the symptoms and needs of the patient change. Family members often provide the bulk of the direct care, and themselves require support, education, and counseling. While we currently lack curative therapies for most patients with malignant brain tumors, careful attention to symptom diagnosis and management can greatly enhance the quality of life of a patient with a brain tumor throughout the course of the illness and at the end of life. PMID- 11854106 TI - Palliative treatment for stroke. AB - Despite continued advances in prevention and treatment, stroke remains the leading cause of neurologic disability. This article reviews the broad spectrum of palliative efforts underway to treat the impairment and disability that results from stroke. Emphasis is placed on the importance of symptom control, increasing compensation, and enhancing residual and recovering function in patients with stroke. New approaches to impairment reduction have produced encouraging results, but disability reduction remains the cornerstone of care. PMID- 11854107 TI - Symptom assessment and management. AB - This article reviews how to assess and manage several symptoms commonly encountered by neurologists who care for patients with advanced illness. Scientifically validated guidelines are reviewed and practical advice is offered on how to manage pain, nausea and vomiting, dyspnea, and respiratory secretions at the end of life. The role of the neurologist as a provider of end of life care is discussed including suggestions for communicating with patients and families. This article concludes with a review of when sedation may be offered within the purview of good palliative care to patients who are imminently dying. PMID- 11854108 TI - Psychiatric dimensions of palliative care. AB - A critically important aspect of palliative care is the prompt recognition and effective treatment of psychiatric complications. Psychiatric syndromes such as depression, anxiety, confusion, suicidal ideation, and wish for hastened death or assisted-suicide occur in a significant percentage of patients with advanced illness. This article reviews their frequency, diagnosis, associated factors, and management to help expand the focus of palliative care beyond pain and physical symptom control. PMID- 11854109 TI - Ethical and legal issues in palliative care. AB - This article reviews the ethical principles underlying palliative care, stressing the importance of respecting patient's rights to withdraw or withhold life sustaining treatment, including artificial hydration and nutrition. There is no ethical or constitutional right to receive physician-assisted suicide or voluntary active euthanasia. This article discusses current ethical controversies in palliative care, including futility, medication dosage and double-effect, terminal sedation, legalization of physician-assisted suicide and euthanasia, and patient refusal of hydration and nutrition. Relevant legal issues are discussed in tandem with the ethical issues. PMID- 11854110 TI - Communication skills in palliative care: a practical guide. AB - Communication skills are frequently regarded as innate and intuitive. Many studies, however, now show that most components of communication techniques can be taught and that these learned skills have an impact on the physician's ability to communicate. This article sets out two fundamental protocols that act as templates or strategies for communication. One, (the C-L-A-S-S strategy) highlights the five central factors in all clinician-patient interviews. The second (the S-P-I-K-E-S protocol) is a variant for the specific task of breaking bad news. PMID- 11854111 TI - Terminal care and bereavement. AB - Care of patients at the end of life requires a high level of clinical vigilance, compassion and skill. The involvement of the patient's primary neurologist in end of-life care and into bereavement can be an invaluable comfort to the patient and family. An understanding of the techniques for assessing and anticipating patient and family needs and knowledge of the resources available is essential if the neurologist is to provide guidance in their care. PMID- 11854116 TI - Pharmacological treatment of coronary artery disease with recombinant fibroblast growth factor-2: double-blind, randomized, controlled clinical trial. AB - BACKGROUND: Single-bolus intracoronary administration of fibroblast growth factor 2 (FGF2) improved symptoms and myocardial function in a phase I, open-label trial in patients with coronary artery disease. We conducted the FGF Initiating RevaScularization Trial (FIRST) to evaluate further the efficacy and safety of recombinant FGF2 (rFGF2). METHODS AND RESULTS: FIRST is a multicenter, randomized, double-blind, placebo-controlled trial of a single intracoronary infusion of rFGF2 at 0, 0.3, 3, or 30 microg/kg (n=337 patients). Efficacy was evaluated at 90 and 180 days by exercise tolerance test, myocardial nuclear perfusion imaging, Seattle Angina Questionnaire, and Short-Form 36 questionnaire. Exercise tolerance was increased at 90 days in all groups and was not significantly different between placebo and FGF-treated groups. rFGF2 reduced angina symptoms as measured by the angina frequency score of the Seattle Angina Questionnaire (overall P=0.035) and the physical component summary scale of the Short-Form 36 (pairwise P=0.033, all FGF groups versus placebo). These differences were more pronounced in highly symptomatic patients (baseline angina frequency score < or =40 or Canadian Cardiovascular Society score of III or IV). None of the differences were significant at 180 days because of continued improvement in the placebo group. Adverse events were similar across all groups, except for hypotension, which occurred with higher frequency in the 30-microg/kg rFGF2 group. CONCLUSIONS: A single intracoronary infusion of rFGF2 does not improve exercise tolerance or myocardial perfusion but does show trends toward symptomatic improvement at 90 (but not 180) days. PMID- 11854117 TI - Increased risk of arrhythmic events in long-QT syndrome with mutations in the pore region of the human ether-a-go-go-related gene potassium channel. AB - BACKGROUND: The hereditary long-QT syndrome is characterized by prolonged ventricular repolarization and a variable clinical course with arrhythmia-related syncope and sudden death. Mutations involving the human ether-a-go-go-related gene (HERG) channel are responsible for the LQT2 form of long-QT syndrome, and in cellular expression studies these mutations are associated with reduction in the rapid component of the delayed rectifier repolarizing current (I(Kr)). We investigated the clinical features and prognostic implications of mutations involving pore and nonpore regions of the HERG channel in the LQT2 form of this disorder. METHODS AND RESULTS: A total of 44 different HERG mutations were identified in 201 subjects, with 14 mutations located in the pore region (amino acid residues 550 through 650). Thirty-five subjects had mutations in the pore region and 166 in nonpore regions. Follow-up extended through age 40 years. Subjects with pore mutations had more severe clinical manifestations of the genetic disorder and experienced a higher frequency (74% versus 35%; P<0.001) of arrhythmia-related cardiac events occurring at earlier age than did subjects with nonpore mutations. Multivariate Cox proportional hazard regression analysis revealed that pore mutations dominated the risk, with hazard ratios in the range of 11 (P<0.0001) for QTc at 500 ms, with a 16% increase in the pore hazard ratio for each 10-ms increase in QTc. CONCLUSION: Patients with mutations in the pore region of the HERG gene are at markedly increased risk for arrhythmia-related cardiac events compared with patients with nonpore mutations. PMID- 11854118 TI - Risk of myocardial infarction and angina in patients with severe peripheral vascular disease: predictive role of C-reactive protein. AB - BACKGROUND: Patients undergoing revascularization procedures for peripheral vascular disease (PVD) have a greatly increased risk for coronary artery disease (CAD) that is predicted only partly by clinical data and cardiovascular risk factors. We investigated whether the prognostic assessment in PVD patients could be improved by preoperative measurements of C-reactive protein (CRP). METHODS AND RESULTS: We assessed clinical and risk factors profiles, Eagle clinical scores, and preoperative CRP serum levels in 51 patients with PVD at Fontaine-Leriche stages II to IV without severe rest ventricular dysfunction or ischemia. During 24 months of follow-up, 17 patients (34%) had fatal (11) or nonfatal (6) myocardial infarction (MI). With univariate logistic regression analysis, only previous history of CAD, Eagle score, and CRP were independently related to MI. At multivariate logistic regression analysis, only CRP values in the upper tertile (<9 mg/L) were significantly associated with MI (P<0.05) and identified 65% of cases. CONCLUSIONS: The high incidence of MI in patients with PVD severe enough to require revascularization is strongly predicted by preprocedural measurements of serum CRP, independent of previous CAD, Eagle score index, and traditional cardiovascular risk factors. These patients may benefit from therapy modulating the inflammatory response. PMID- 11854119 TI - Reduction of inflammatory cytokine concentrations and improvement of endothelial functions in obese women after weight loss over one year. AB - BACKGROUND: Visceral fat is a key regulator site for the process of inflammation, and atherosclerotic lesions are essentially an inflammatory response. METHODS AND RESULTS: Fifty-six healthy premenopausal obese women (age range 25 to 44 years, body mass index 37.2+/-2.2, waist to hip ratio range 0.78 to 0.92) and 40 age matched normal weight women were studied. Compared with nonobese women, obese women had increased basal concentrations of tumor necrosis factor-alpha (TNF alpha, P<0.01), interleukin-6 (IL-6, P<0.01), P-selectin (P<0.01), intercellular adhesion molecule-1 (ICAM-1, P<0.02), and vascular adhesion molecule-1 (VCAM-1, P<0.05). Vascular responses to L-arginine (3 g IV), the natural precursor of nitric oxide, were impaired in obese women: reductions in mean blood pressure (P<0.02), platelet aggregation to adenosine diphosphate (P<0.05), and blood viscosity (P<0.05) were significantly lower as compared with those in the nonobese group. Concentrations of TNF-alpha and IL-6 were related (P<0.01) to visceral obesity, as well as to adhesin levels and responses to L-arginine. After 1 year of a multidisciplinary program of weight reduction (diet, exercise, behavioral counseling), all obese women lost at least 10% of their original weight (9.8+/-1.5 kg, range 7.5 to 13 kg). Compared with baseline, sustained weight loss was associated with reduction of cytokine (P<0.01) and adhesin (P<0.02) concentrations and with improvement of vascular responses to L-arginine. CONCLUSION: In obese women, endothelial activation correlates with visceral body fat, possibly through inappropriate secretion of cytokines. Weight loss represents a safe method for downregulating the inflammatory state and ameliorating endothelial dysfunction in obese women. PMID- 11854120 TI - Fibrillin-1 genotype is associated with aortic stiffness and disease severity in patients with coronary artery disease. AB - BACKGROUND: Elevated pulse pressure is associated strongly with adverse cardiovascular outcome; however, the genetic basis of this condition is unknown. This study examined whether genotypic variation in the extracellular matrix protein fibrillin-1, the Marfan gene, was associated with aortic stiffening and therefore could contribute to cardiovascular risk associated with pulse pressure elevation in coronary disease. METHODS AND RESULTS: Patients (n=145; 113 men), 62+/-9 years of age (mean+/-SD), with angiographically confirmed coronary disease, were studied. Carotid applanation tonometry was used to assess central blood pressures, and in conjunction with Doppler velocimetry, to assess aortic input and characteristic impedance. Fibrillin-1 genotype was characterized by a variable nucleotide tandem repeat and 2 single-nucleotide polymorphisms. The variable nucleotide tandem repeat was a good predictor of underlying haplotypes with 3 genotypes (2-2, 2-4, and 2-3) accounting for 86% of the population. The 2 3 genotype had higher input impedance (P=0.002), characteristic impedance (P=0.005), and carotid pulse pressure (P=0.002) compared with the 2-2 and 2-4 genotypes. Disease severity assessed by previous angioplasties and the number of patients with a stenosis >90% was also greater in the 2-3 genotype. Furthermore, in a multivariate analysis, fibrillin-1 genotype and central pulse pressure were independent of conventional risk factors in determining coronary disease severity. There was no difference in age, sex ratio, body mass index, smoking status, cholesterol level, or medication among the 3 genotypes. CONCLUSIONS: Although a causative link has not been shown, these data are consistent with an important role for fibrillin-1 genotype in cardiovascular risk associated with large-artery stiffening and pulse pressure elevation in individuals with coronary disease. PMID- 11854121 TI - Advanced glycation end products activate endothelium through signal-transduction receptor RAGE: a mechanism for amplification of inflammatory responses. AB - BACKGROUND: The products of nonenzymatic glycation and oxidation of proteins, the advanced glycation end products (AGEs), form under diverse circumstances such as aging, diabetes, and kidney failure. Recent studies suggested that AGEs may form in inflamed foci, driven by oxidation or the myeloperoxidase pathway. A principal means by which AGEs alter cellular properties is through interaction with their signal-transduction receptor RAGE. We tested the hypothesis that interaction of AGEs with RAGE on endothelial cells enhances vascular activation. METHODS AND RESULTS: AGEs, RAGE, vascular cell adhesion molecule-1, intercellular adhesion molecule-1, and E-selectin are expressed in an overlapping manner in human inflamed rheumatoid synovia, especially within the endothelium. In primary cultures of human saphenous vein endothelial cells, engagement of RAGE by heterogeneous AGEs or Nepsilon(carboxymethyl)lysine-modified adducts enhanced levels of mRNA and antigen for vascular cell adhesion molecule-1, intercellular adhesion molecule-1, and E-selectin. AGEs increased adhesion of polymorphonuclear leukocytes to stimulated endothelial cells in a manner reduced on blockade of RAGE. CONCLUSIONS: AGEs, through RAGE, may prime proinflammatory mechanisms in endothelial cells, thereby amplifying proinflammatory mechanisms in atherogenesis and chronic inflammatory disorders. PMID- 11854122 TI - Value of stress myocardial perfusion single photon emission computed tomography in patients with normal resting electrocardiograms: an evaluation of incremental prognostic value and cost-effectiveness. AB - BACKGROUND: The incremental value and cost-effectiveness of stress single photon emission computed tomography (SPECT) is of unclear added value over clinical and exercise treadmill testing data in patients with normal resting ECGs, a patient subset known to be at relatively lower risk. METHODS AND RESULTS: We identified 3058 consecutive patients who underwent exercise dual isotope SPECT, who on follow-up (mean, 1.6+/-0.5 years; 3.6% lost to follow-up) were found to have 70 hard events (2.3% hard-event rate). Survival analysis used a Cox proportional hazards model, and cost-effectiveness was determined by the cost per hard event identified by strategies with versus without the use of SPECT. In this cohort, a normal study was associated with an exceedingly low hard-event rate (0.4% per year) that increased significantly as a function of the SPECT result. After adjusting for pre-SPECT information, exercise stress SPECT yielded incremental value for the prediction of hard events (chi2 52 to 85, P<0.001) and significantly stratified patients. In patients with intermediate to high likelihood of coronary artery disease after exercise treadmill testing, a cost effectiveness ratio of $25 134 per hard event identified and a cost of $5417 per reclassification of patient risk were found. Subset analyses revealed similar prognostic, and cost results were present in men, women, and patients with and without prior histories of coronary artery disease. CONCLUSIONS: Stress SPECT yields incremental prognostic value and enhanced risk stratification in patients with normal resting ECGs in a cost-effective manner. These findings are opposite those of previous studies examining anatomic end points in this same population and thus, if confirmed, have significant implications for patient management. PMID- 11854124 TI - Effects of selective autonomic blockade on T-wave alternans in humans. AB - BACKGROUND: T-wave alternans (TWA) is an important noninvasive measure of ventricular arrhythmia vulnerability. This study tested the hypothesis that the autonomic nervous system influences TWA measurement in high-risk subjects with coronary artery disease. METHODS AND RESULTS: T-wave alternans was measured in 60 patients with coronary artery disease, left ventricular dysfunction, and inducible sustained ventricular tachycardia during electrophysiological studies. All patients had TWA measured at baseline with atrial pacing at 100 bpm (600 ms), 109 bpm (550 ms), and 120 bpm (500 ms). After a 10-minute recovery period, TWA was measured again after sympathetic blockade (esmolol, n=20), parasympathetic blockade (atropine, n=20), or no intervention (control subjects, n=20). The prevalence of significant TWA was unchanged compared with baseline after atropine infusion and in the control group. In contrast, the amplitude of TWA in the vector magnitude lead was significantly reduced after esmolol infusion (P<0.001), and the number of positive TWA tests was reduced by 50% (70% versus 35%, P<0.05). CONCLUSIONS: Our findings have important implications for the use of TWA to risk stratify patients for life-threatening ventricular arrhythmias and provide a new potential mechanism for the reduction in sudden cardiac death conferred by beta blockers among patients with coronary artery disease and congestive heart failure. PMID- 11854123 TI - Relationship between treatment-induced changes in left ventricular mass and blood pressure in black african hypertensive patients: results of the Baragwanath Trial. AB - BACKGROUND: In a single-center study, we compared to what extent changes in conventional and ambulatory blood pressure (BP) predicted regression of left ventricular mass (LVM) index in response to antihypertensive treatment in previously untreated and treated patients with sustained hypertension. METHODS AND RESULTS: We enrolled 173 black African patients who, off treatment, had a daytime diastolic BP ranging from 90 to 114 mm Hg. Antihypertensive drugs were titrated and combined to reduce the daytime diastolic BP below 90 mm Hg. Echocardiograms were obtained at baseline and follow-up. Mean systolic/diastolic clinic BP, 24-hour BP, and LVM index were similar in previously untreated (n=64) and previously treated (n=109) patients and averaged 171/102 mm Hg, 151/97 mm Hg, and 118 g/m2, respectively. At 4 months, these values had decreased (P<0.001) by 26/12 mm Hg, 23/14 mm Hg, and 14 g/m2 in previously untreated patients and by 22/9 mm Hg, 21/13 mm Hg, and 19 g/m2 in previously treated patients. In the previously untreated patients, the regression in LVM index correlated to a similar degree (P=0.09) with the decreases in the conventional (r=0.34; P=0.005) and the 24-hour (r=0.26; P=0.04) systolic BP. In the previously treated patients, the corresponding correlations were 0.02 (P=0.82) and -0.10 (P=0.32), respectively. Compared with the 24-hour systolic BP, automated oscillometric measurements of systolic BP obtained at the clinic yielded similar results. CONCLUSIONS: In previously untreated patients with sustained hypertension followed at a single center, reductions in clinic and ambulatory systolic pressure in response to antihypertensive treatment equally predicted the regression in LVM index. PMID- 11854125 TI - Maternal anti-Ro and anti-La antibody-associated endocardial fibroelastosis. AB - BACKGROUND: Maternal anti-Ro and anti-La antibodies are associated with congenital heart block (CHB). Although endocardial fibroelastosis (EFE) has been described in isolated cases of autoantibody-mediated CHB, the natural history and pathogenesis of this disease are poorly understood. METHODS AND RESULTS: We retrospectively reviewed the clinical history, echocardiography, and pathology of fetuses and children with EFE associated with CHB born to mothers positive for anti-Ro or anti-La antibodies at 5 centers. Thirteen patients were identified, 6 with a prenatal and 7 with a postnatal diagnosis. Six mothers were positive for anti-Ro and anti-La antibodies, and 7 were positive for anti-Ro antibodies only. Only 1 mother had autoimmune disease. Severe ventricular dysfunction was seen in all fetal and postnatal cases. Four fetal and 3 postnatal cases had EFE at initial presentation. However, 2 fetal and 4 postnatal cases developed EFE 6 to 12 weeks and 7 months to 5 years from CHB diagnosis, respectively, even despite ventricular pacing in 6 postnatal cases. Eleven (85%) either died (n=9) or underwent cardiac transplantation (n=2) secondary to the EFE. Pathologic assessment of the explanted heart, available in 10 cases, revealed moderate to severe EFE in 7 and mild EFE in 3 cases, predominantly involving the left ventricle. Immunohistochemistry in 4 cases (including 3 fetuses) demonstrated deposition of IgG in 4 and IgM in 3 and T-cell infiltrates in 3 cases, suggesting an immune response by the affected fetus or child. CONCLUSIONS: EFE occurs in the presence of autoantibody-mediated CHB despite adequate ventricular pacing. Autoantibody-associated EFE has a very high mortality rate, whether developing in fetal or postnatal life. PMID- 11854126 TI - Cigarette smoke exposure and hypercholesterolemia increase mitochondrial damage in cardiovascular tissues. AB - BACKGROUND: A shared feature among cardiovascular disease risk factors is increased oxidative stress. Because mitochondria are susceptible to damage mediated by oxidative stress, we hypothesized that risk factors (secondhand smoke and hypercholesterolemia) are associated with increased mitochondrial damage in cardiovascular tissues. METHODS AND RESULTS: Atherosclerotic lesion formation, mitochondrial DNA damage, protein nitration, and specific activities of mitochondrial proteins in cardiovascular tissues from age-matched C57 and apoE(-/ ) mice exposed to filtered air or secondhand smoke were quantified. Both secondhand smoke and hypercholesterolemia were associated with significantly increased mitochondrial DNA damage and protein nitration. Tobacco smoke exposure also resulted in significantly decreased specific activities of mitochondrial enzymes. The combination of secondhand smoke and hypercholesterolemia resulted in increased atherosclerotic lesion formation and even greater levels of mitochondrial damage. CONCLUSIONS: These data are consistent with the hypothesis that cardiovascular disease risk factors cause mitochondrial damage and dysfunction. PMID- 11854127 TI - Effect of hypercholesterolemia on expression and function of vascular soluble guanylyl cyclase. AB - BACKGROUND: Vasorelaxation to endothelial NO is mediated by activation of soluble guanylyl cyclase (sGC) and impaired by hypercholesterolemia in animals and humans. We investigated whether hypercholesterolemia impacts expression and function of sGC. METHODS AND RESULTS: White New Zealand rabbits (n=10 per group) received a standard diet for 16 weeks (SD16) (n=20) or 32 weeks (SD32) and a cholesterol diet (7.5 g/kg) for 16 weeks (CD16) (n=20) or 32 weeks (CD32), respectively. Another group received cholesterol diet for 16 weeks followed by standard diet for 16 weeks (CD/SD). Aortic expression of the alpha1-subunit of sGC (sGC-alpha1) and beta1-subunit of sGC (sGC-beta1) was assessed by Western blot. Function was measured by aortic relaxation to S-Nitroso-N-acetyl-D, L penicillamine (SNAP) and sGC activity in aortic cytosols. Hypercholesterolemia induced an upregulation of sGC-beta1 in CD16 (3.5+/-0.4-fold, P<0.001 versus SD16) and CD32 (4.0+/-0.4-fold, P<0.001 versus SD32). A similar increase was found for sGC-alpha1. In striking contrast, basal and NO-stimulated sGC activities in aortic cytosols of CD16 were only slightly enhanced (1.4-fold, P<0.05). Furthermore, the vasodilator potency of SNAP (EC50 in -logM) was 10-fold lower in CD16 (6.76a+/-0.09) than in SD16 (7.66+/-0.14, P<0.01). The increase of sGC expression was completely reversible, as indicated by comparable sGC-beta1 amounts in SD32 and CD/SD (1.2+/-0.1-fold, P>0.05). Immunohistochemical analysis suggests that a great portion of the overexpressed sGC is located in intimal lesions. Additional experiments showed that increased vascular superoxide production induced by 6-anilino-5,8-quinolinedione (LY85385) reduces sGC-activity but increases sGC-expression. CONCLUSIONS: These results suggest that hypercholesterolemia induces a reversible overexpression of a dysfunctional vascular sGC, which may contribute to the pathogenesis of atherosclerosis. PMID- 11854128 TI - Porphyromonas gingivalis infection accelerates the progression of atherosclerosis in a heterozygous apolipoprotein E-deficient murine model. AB - BACKGROUND: Current epidemiological data suggest that dental infections affecting tooth-supporting tissues (periodontitis) can disseminate into the systemic circulation and thereby contribute to atherosclerosis progression. To test this hypothesis, we investigated the effect of repeated systemic inoculations with Porphyromonas gingivalis (Pg), a putative periodontal pathogen, on the progression of atherosclerosis in heterozygous apolipoprotein E-deficient (ApoE(+/-)) mice. METHODS AND RESULTS: Ten-week-old, male ApoE(+/-) mice fed either a high-fat diet or regular chow were inoculated intravenously with live Pg (10(7) CFU) or vehicle once per week for 10, 14, or 24 consecutive weeks. Histomorphometry of plaque cross-sectional area in the proximal aortas, en face measurement of plaque area over the aortic trees, Pg 16S ribosomal DNA amplification with polymerase chain reaction, ELISA for systemic proinflammatory mediators, and immunolocalization of macrophages in the proximal aorta were performed. Atherosclerotic lesions of the proximal aortas and aortic trees were more advanced in Pg-challenged animals than in vehicle control animals and occurred earlier (at 10 weeks) when no lesions were apparent in control animals. At 24 weeks after inoculation, proximal aortic lesion size quantified by histomorphometry was 9-fold greater in chow-fed mice inoculated with Pg than in noninoculated mice (P<0.001) and was 2-fold greater in Pg-inoculated versus noninoculated high-fat diet-fed mice (P<0.001); all atherosclerotic lesions were macrophage-rich. Pg ribosomal DNA was found in the aortas, livers, and hearts 24 weeks after inoculation. CONCLUSIONS: These results provide evidence that long term systemic challenge with Porphyromonas gingivalis, an oral pathogen, can accelerate atherogenic plaque progression. PMID- 11854129 TI - Fluvastatin, a 3-hydroxy-3-methylglutaryl coenzyme a reductase inhibitor, attenuates left ventricular remodeling and failure after experimental myocardial infarction. AB - BACKGROUND: Short-term administration of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors, statins, has been shown to attenuate ischemia reperfusion injury. However, the effects of long-term administration of statins on left ventricular (LV) remodeling and failure after myocardial infarction remain unknown. METHODS AND RESULTS: Mice were subjected to coronary artery ligation and were treated for 4 weeks with vehicle or fluvastatin (10 mg/kg per day PO). Fluvastatin increased survival (61% versus 86%; P<0.05) without affecting the infarct size (52+/-2% versus 49+/-3%; P=NS). Fluvastatin not only attenuated LV dilatation but also decreased LV end-diastolic pressure and lung weight. Furthermore, it reduced cardiac myocyte hypertrophy and interstitial fibrosis of the noninfarcted LV and also improved LV ejection performance. LV matrix metalloproteinase (MMP)-2 and MMP-13 were increased in myocardial infarction, which was attenuated in fluvastatin-treated mice. CONCLUSIONS: Fluvastatin increased survival in a murine model of postinfarct heart failure, which was associated with the amelioration of LV structural remodeling and contractile failure. Moreover, these effects were associated with the attenuation of increased MMP activity. Thus, long-term treatment with fluvastatin might be beneficial also in patients with heart failure and might improve their long-term survival. PMID- 11854130 TI - Magnetic resonance--guided coronary artery stent placement in a swine model. AB - BACKGROUND: Magnetic resonance (MR)--guided coronary artery stent placement is a challenging vascular intervention because of the small size of the coronary arteries combined with incessant motion during the respiratory and cardiac cycles. These obstacles necessitate higher temporal and higher spatial resolution real-time MR imaging techniques when compared with interventional peripheral MR angiography. METHODS AND RESULTS: A new, ultrafast, real-time MR imaging technique that combines steady-state free precession (SSFP) for high signal-to noise ratio and radial k-space sampling (rSSFP) for motion artifact suppression was implemented on a 1.5-T clinical whole-body interventional MR scanner. The sliding window reconstruction technique yielded a frame rate of 15/s allowing for data acquisition during free breathing and without cardiac triggering. Eleven balloon-expandable stainless steel coronary stents were placed in both coronary arteries of 7 pigs (40 to 70 kg body weight) using a nitinol guidewire and passive device visualization. Position of the coronary stents was controlled by a navigator-gated free-breathing ECG-triggered three-dimensional SSFP coronary MRA sequence and confirmed visually on the ex vivo heart. The presented real-time MR imaging sequence reliably allowed for high-quality coronary MR fluoroscopy without motion artifacts in all pigs. Ten of 11 coronary stents were correctly placed under MR guidance. One stent dislodged proximally from the left main coronary artery because of too-small balloon size. Stent dislocation was correctly predicted during real-time MR imaging. CONCLUSION: The presented approach allows for real-time MR-guided coronary artery stent placement in a swine model. PMID- 11854131 TI - Atrio-sinus interaction demonstrated by blockade of the rapid delayed rectifier current. AB - BACKGROUND: Proper pacemaking of the heart requires a specific organization of the sinoatrial (SA) node. The SA node drives the surrounding atrium but needs to be protected from its hyperpolarizing influence, which tends to suppress pacemaker activity. It has been suggested that the hyperpolarizing atrial influence is minimal at the site of the central nodal area. METHODS AND RESULTS: Atrio-sinus interaction was assessed by specific depolarization of the SA node by blocking the HERG-encoded rapid delayed rectifier current (I(K,r)) with the drug E-4031. In the SA node, E-4031 (1 micromol/L) changed action potential configuration drastically but never resulted in pacemaker arrest. In the atrium, E-4031 did not affect the membrane resting potential, thereby leaving the normal hyperpolarizing load on the SA node intact. When the SA node was sectioned into strips and subsequently separated from the atrium, spontaneous electrical activity of the strip containing the primary pacemaker ceased on I(K,r) blockade. When not separated from the atrium, I(K,r) blockade never resulted in pacemaker arrest. A similar effective atrio-sinus interaction was demonstrated in computer simulations. CONCLUSIONS: Our results demonstrate that the atrium provides an effective hyperpolarizing load on the central SA nodal area and is at least one of the controlling mechanisms for normal pacemaking function. The present study can be of help in understanding why patients with long-QT2 syndrome secondary to a mutation in HERG do not show sinus arrest. PMID- 11854132 TI - New tools for coronary risk assessment: what are their advantages and limitations? AB - The concept of risk assessment and reduction, introduced initially by the Framingham Heart Study and refined in other models, forms the cornerstone of preventive cardiology. Risk factor assessment determines the therapeutic strategy, because the intensity of preventive intervention is tailored to the patient's risk of coronary heart disease. The conventional risk factors for coronary heart disease include elevated serum total cholesterol and LDL cholesterol, low HDL cholesterol, elevated blood pressure, cigarette smoking, diabetes, vascular disease, menopausal status (women only), and age. Aggressive risk factor reduction, formerly used exclusively in secondary prevention, may be pivotal to optimal patient management in high-risk primary prevention. A number of noninvasive imaging modalities have the potential to measure and to monitor atherosclerosis in asymptomatic individuals and include exercise ECG testing, electron beam computed tomography, magnetic resonance coronary angiography, positron emission tomography, ankle-brachial index, and B-mode ultrasound. Novel serum markers, including C-reactive protein and homocysteine, have the ability to gauge risk in the individual patient. Systemic therapy for risk reduction in primary prevention may be preferable to local therapy (eg, angioplasty and bypass) and may more effectively prevent acute coronary events than these more invasive approaches. PMID- 11854133 TI - Prevention of coronary heart disease by diet and lifestyle: evidence from prospective cross-cultural, cohort, and intervention studies. PMID- 11854134 TI - Images in cardiovascular medicine. Flash pulmonary edema heralding renal artery stenosis. PMID- 11854135 TI - Postoperative dislocation of the heart after pneumonectomy. PMID- 11854136 TI - Sustain weight loss to downregulate inflammation. PMID- 11854137 TI - Structural characterization of in vivo rat glutathione adducts and a hydroxylated metabolite of simvastatin hydroxy acid. AB - Simvastatin hydroxy acid (SVA), the pharmacologically active form of simvastatin (SV), is a potent inhibitor of 3-hydroxy-3-methylglutaryl (HMG)-coenzyme A reductase and is formed on hydrolysis of the orally administered SV. In this article, we report the structural characterization of two new dihydroxy glutathione adducts and a trihydroxy derivative of SVA, all found in rat bile. Metabolite I is 5'beta,6'beta-dihydroxy-4'a(alpha)-glutathione-SVA, and metabolite II is a pentanoic acid derivative of metabolite I. The two identified GSH conjugates accounted for 16 and 9% in males and 11 and 5% in females of the total radioactivity (metabolites I and II, respectively). Metabolite III is 3',5'beta,6'beta-dihydrotriol-SVA and accounts for 2% (male) and 4% (female) of the total dose in rats. Of these three newly identified metabolites, only metabolite III was also observed in dog bile. PMID- 11854138 TI - Endomorphins, Met-enkephalin, Tyr-MIF-1, and the P-glycoprotein efflux system. AB - The P-glycoprotein (P-gp) transport system, responsible for the efflux of many therapeutic drugs out of the brain, recently has been shown to transport the endogenous brain opiate endorphin. We used P-gp knockout mice (Mdr1a) and their controls to determine where P-gp is involved in the saturable efflux systems of four other endogenous opiate-modulating peptides across the blood-brain barrier (BBB). After injection of endomorphin-1 (Tyr-Pro-Trp-Phe-NH(2)), endomorphin-2 (Tyr-Pro-Phe-Phe-NH(2)), Met-enkephalin (Tyr-Gly-Gly-Phe-Met-OH), and Tyr-MIF-1 (Tyr-Pro-Leu-Gly-NH(2)) into the lateral ventricle of the mouse brain, residual radioactivity was measured at 0, 2, 5, 10, and 20 min later. The results showed no difference in the disappearance of any of these peptides from the brains of knockout mice compared with their controls. This demonstrates that unlike endorphin and morphine, P-gp does not seem to be required for the brain-to-blood transport of the endomorphins, Met-enkephalin, or Tyr-MIF-1 across the BBB. PMID- 11854139 TI - (+)-N-3-Benzyl-nirvanol and (-)-N-3-benzyl-phenobarbital: new potent and selective in vitro inhibitors of CYP2C19. AB - Highly potent and selective CYP2C19 inhibitors are not currently available. In the present study, N-3-benzyl derivatives of nirvanol and phenobarbital were synthesized, their respective (+)- and (-)-enantiomers resolved chromatographically, and inhibitor potencies determined for these compounds toward CYP2C19 and other human liver cytochromes P450 (P450s). (-)-N-3-Benzyl phenobarbital and (+)-N-3-benzyl-nirvanol were found to be highly potent, competitive inhibitors of recombinant CYP2C19, exhibiting K(i) values of 79 and 250 nM, respectively, whereas their antipodes were 20- to 60-fold less potent. In human liver preparations, (-)-N-3-benzyl-phenobarbital and (+)-N-3-benzyl nirvanol inhibited (S)-mephenytoin 4'-hydroxylase activity, a marker for native microsomal CYP2C19, with K(i) values ranging from 71 to 94 nM and 210 to 280 nM, respectively. At single substrate concentrations of 0.3 microM [(-)-N-3-benzyl phenobarbital] and 1 microM [(+)-N-3-benzyl-nirvanol] that were used to examine inhibition of a panel of cDNA-expressed P450 isoforms, neither CYP1A2, 2A6, 2C8, 2C9, 2D6, 2E1, nor 3A4 activities were decreased by greater than 16%. In contrast, CYP2C19 activity was inhibited approximately 80% under these conditions. Therefore, (+)-N-3-benzyl-nirvanol and (-)-N-3-benzyl-phenobarbital represent new, highly potent and selective inhibitors of CYP2C19 that are likely to prove generally useful for screening purposes during early phases of drug metabolism studies with new chemical entities. PMID- 11854140 TI - Increase in rat liver UDP-glucuronosyltransferase mRNA by microsomal enzyme inducers that enhance thyroid hormone glucuronidation. AB - Treatment of rats with the microsomal enzyme inducers pregnenolone-16alpha carbonitrile (PCN), 3-methylcholanthrene (3-MC), and Aroclor 1254 [PCB (polychlorinated biphenyl)] has been shown to decrease circulating levels of thyroid hormones as well as increase microsomal glucuronidation of thyroxine (T(4)). In addition, PCN increases triiodothyronine (T(3)) uridine diphosphate glucuronosyltransferase (UGT) activity. Members of the UGT1A family are believed to glucuronidate T(4), specifically UGT1A1 and UGT1A6, whereas the UGT2 family is believed to glucuronidate T(3), namely UGT2B2. The purpose of this study was to determine whether the aforementioned microsomal enzyme inducers increase the mRNAs that encode these and other UGT enzymes in rat liver. Male Sprague-Dawley rats were fed a control diet or a diet containing PCN (1000 ppm), 3-MC (250 ppm), or PCB (100 ppm) for 7 days, at which time livers were collected. Increases in mRNA were detected by QuantiGene branched DNA signal amplification. A 3-fold increase in UGT1A1 mRNA was produced by PCN in addition to increases in UGT1A2 (4 fold) and UGT1A5 (2-fold) mRNA. PCN affected neither UGT2B2 nor any other UGT2B mRNA level. 3-MC and PCB increased UGT1A6 mRNA 6- and 4-fold, respectively. 3-MC and PCB each increased UGT1A7 mRNA 4-fold but did not significantly increase any other UGT mRNAs. These findings suggest that PCN enhances T(4) UGT activity by increased expression of UGT1A1 and that 3-MC and PCB enhance T(4) UGT activity by increased expression of UGT1A6. These findings also suggest that increased T(3) UGT activity produced by PCN is due to a mechanism other than increased transcription of UGT2B2, possibly increased UGT2B2 protein or induction of another UGT enzyme. PMID- 11854141 TI - Urinary naphthalene mercapturates as biomarkers of exposure and stereoselectivity of naphthalene epoxidation. AB - Previous work has shown that the rate and stereochemistry of naphthalene epoxidation correlates with differences in susceptibility to cytotoxicity. The development of methods for measuring epoxide formation in vivo could provide a marker for assessing events critical to naphthalene cytotoxicity that are applicable to humans. Here, urinary diastereomeric mercapturates have been measured in mice (susceptible) and rats (nonsusceptible) after intraperitoneal administration (1.56-200 mg/kg) or inhalation exposures (0.8-110 ppm, 4 h) to naphthalene. No significant differences were observed in the percentage of the dose eliminated as mercapturate in urine between mice (25-34%) and rats (24-35%) or at varying doses after i.p. administration. The amounts of urinary mercapturate after 4-h exposures were considerably greater in mice than rats. In mice, the ratio of diastereomeric mercapturates derived from the 1R,2S- to 1S,2R epoxide was 1:1 at low doses (1-3 mg/kg), increased to 3:1 at intermediate doses (50 mg/kg), and decreased to 2:1 at high doses (100 and 200 mg/kg). In rats, these ratios remained less than 1:1 at all doses. After inhalation, ratios were 5 to 6:1 at low concentrations (less than 15 ppm) and decreased to 3:1 at higher concentrations (15-100 ppm) in mice, whereas in rats, the ratios were 1:1 or less for all concentrations. These studies show that mercapturates provide good assessments of internal dose, that there are not significant differences between mice and rats in the percentage eliminated as mercapturate but that the ratios of mercapturates derived from the 1R,2S- versus 1S,2R-epoxide differ markedly and are consistent with previous in vitro metabolism studies. PMID- 11854142 TI - The pharmacokinetic determinants of the frequency and pattern of intravenous cocaine self-administration in rats by pharmacokinetic modeling. AB - We investigated the pharmacokinetic determinants of the frequency of intravenous cocaine self-administration in 2.5-h sessions. Two groups of rats were implanted with dual catheters that permitted cocaine infusion and blood sampling via the femoral and jugular vein catheters, respectively. Half of the animals in each group self-administered one of the two cocaine unit doses (0.5 and 1 mg/kg/infusion) by pressing a lever under a continuous schedule of reinforcement. To monitor serum cocaine concentrations, the remaining animals received concurrent, response-independent infusions whenever the matched animals self administered cocaine infusions. Multiple concentration-time data in two successive self-administrations were determined to monitor the extent of fluctuation in concentrations by pharmacokinetic modeling. Behavioral analyses revealed the higher unit dose (1 mg/kg) resulted in less frequent cocaine self administration, and a longer interinfusion interval, whereas the total doses were similar for the two groups (24.5-27.0 mg/kg/2.5 h). Cocaine decayed biexponentially. Both the values of clearance and terminal elimination rate constant for the self-administration paradigm were significantly greater than those after the bolus cocaine dosing series (0.5 and 1 mg/kg, separated by 3 days). The regularity in cocaine self-administration produced relatively stable serum cocaine concentrations that oscillated between maximum (C(max)) and minimum (C(min)) values regardless of dose size and interinfusion interval. Although the C(max) for the 1-mg/kg unit dose (1.47 microg/ml) was significantly higher than that for the 0.5-mg/kg dose (0.82 microg/ml), the C(min) values between the groups approximated each other (0.28, and 0.34 microg/ml, respectively). Hence, the C(min) is the determinant of the initiation of the next drug-taking behavior. PMID- 11854144 TI - Characterization by liquid chromatography-nuclear magnetic resonance spectroscopy and liquid chromatography-mass spectrometry of two coupled oxidative-conjugative metabolic pathways for 7-ethoxycoumarin in human liver microsomes treated with alamethicin. AB - The microsomal metabolism of 7-ethoxycoumarin (7-EC) was investigated using liquid chromatography (LC)-NMR and liquid chromatography-mass spectrometry (LC MS) to characterize the coupling of oxidative-conjugative metabolism events. Within microsomes, cytochromes P450 (P450s) and UDP-glucuronosyltransferases (UGTs) are spatially disparate, each having surface and luminal localization, respectively. To optimize cofactor and substrate transit to UGT without compromising P450 activity, the pore-forming peptide alamethicin was used for microsomal perforation. Aqueous extracts of microsomal incubations containing NADPH and UDP-glucuronic acid were injected for LC-NMR and LC-MS analysis. The analytical complementarity of LC-NMR and LC-MS permitted the identification of four metabolites (M1 to M4). The metabolites M1 and M2 are novel microsomal metabolites for 7-EC, consistent with 3-hydroxylation and subsequent glucuronidation, respectively. Metabolites M3 and M4 were 7-hydroxycoumarin (7 HC) and 7-HC glucuronide, respectively. Viewed collectively, these results illustrate the utility of alamethicin in the examination of coupled oxidative conjugative metabolism and the synergy of LC-NMR and LC-MS in metabolite identification. PMID- 11854143 TI - Induction of CYP1A1 and CYP1B1 in T-47D human breast cancer cells by benzo[a]pyrene is diminished by arsenite. AB - Polycyclic aromatic hydrocarbons (PAHs) and metals are often environmental cocontaminants, yet there have been relatively few studies of combined effects of PAHs and metals on cytochrome P450 (P450)-catalyzed metabolism. We examined the effects of NaAsO(2) in combination with benzo[a]pyrene (BAP) on CYP1A1 and CYP1B1 in T-47D human breast cancer cells by using estrogen metabolism as a probe of their activities. Exposure to BAP caused elevated rates of the 2- and 4 hydroxylation pathways of estrogen metabolism, indicating induction of both CYP1A1, an estradiol 2-hydroxylase, and CYP1B1, an estradiol 4-hydroxylase. BAP induced metabolism peaked 9 to 16 h after exposure and returned to near-basal levels by 48 h. Concentration-response studies showed maximal induction of the 2- and 4-hydroxylation pathways at 3 microM BAP; higher levels caused reduced rates of metabolism due to inhibition of CYP1A1 and CYP1B1. NaAsO(2) caused pronounced decreases in the induction of CYP1A1 and CYP1B1 by 3 microM BAP because cotreatment with 10 microM NaAsO(2) inhibited the rates of the 2- and 4 hydroxylation pathways by 86 and 92%, respectively. Western immunoblots showed diminished levels of BAP-induced CYP1A1 by coexposure to NaAsO(2). The levels of the CYP1A1 and CYP1B1 mRNAs induced by BAP were not significantly affected by coexposure to NaAsO(2); however, heme oxygenase 1 mRNA levels were markedly induced by coexposure to BAP and NaAsO(2). These results indicate a post transcriptional inhibitory effect of arsenite on the expression of CYP1A1 and CYP1B1 in T-47D cells, possibly resulting from reduced heme availability. PMID- 11854145 TI - Incorporating measures of variability and uncertainty into the prediction of in vivo hepatic clearance from in vitro data. AB - The existing procedures for quantitative in vitro-in vivo clearance prediction can be significantly biased either by totally neglecting the existing variability and uncertainty by using mean parameter values or by implementing Monte Carlo simulation with statistical distribution of the parameters reconstructed from very small sets of data. The aim of the present study is to develop a methodology for the prediction of in vivo hepatic clearance in the presence of semiquantitative or qualitative data and accounting for the existing uncertainty and variability. The method consists of two steps: 1) transformation of the information available into fuzzy sets (fuzzification); and 2) computation of the in vivo clearance using arithmetic operations with fuzzy sets. To illustrate the approach, rat hepatocyte and microsomal data for eight benzodiazepine compounds are used. A comparison with a standard Monte Carlo procedure is made. The methodology proposed can be used when Monte Carlo simulation may be biased or cannot be implemented. The obtained fuzzy in vivo clearance can be used subsequently in fuzzy simulations of pharmacokinetic models. PMID- 11854146 TI - Postnatal expression and induction by pregnenolone-16alpha-carbonitrile of the organic anion-transporting polypeptide 2 in rat liver. AB - Newborn rats are more sensitive to the toxic effects of cardiac glycosides than are adult rats. This is associated with a decreased ability to remove cardiac glycosides from blood into the liver. Pregnenolone-16alpha-carbonitrile (PCN), a prototypical rodent CYP3A inducer and pregnane-X-receptor (PXR) ligand, stimulates the hepatic clearance of cardiac glycosides in newborn rats, which results in decreased toxicity of the cardiac glycosides. The mechanism responsible for this phenomenon is not clear; however, if elucidated, it would help in understanding and preventing potential drug-drug interactions. The recently cloned rat organic anion-transporting polypeptide 2 (oatp2) (Slc21a5) is a sinusoidal hepatic uptake transporter, with very high affinities for cardiac glycosides, and thus it was hypothesized that rat oatp2 increases during postnatal development and is inducible by PCN. In the present study, livers were removed from Sprague-Dawley rats from postnatal days (pnd) 0 to 45, in 5-day increments; as well as from pnd 10 to 90, in 10-day increments, after PCN (75 mg/kg i.p., for 4 days) or corn oil (vehicle for PCN) treatment. The protein and mRNA levels of rat oatp2 were determined by Western blot analysis and branched DNA signal amplification technique, respectively. Expression of rat oatp2 protein and mRNA increased gradually during postnatal development. PCN treatment increased liver to body weight ratio in both genders, and dramatically accelerated the maturation of hepatic oatp2 protein and mRNA levels. In summary, rat oatp2 undergoes age-dependent and chemical regulation during postnatal development, and is a potential target for drug-drug and age-drug interactions. PMID- 11854147 TI - In vitro human metabolism and interactions of repellent N,N-diethyl-m-toluamide. AB - Oxidative metabolism of the insect repellent N,N-diethyl-m-toluamide (DEET) by pooled human liver microsomes (HLM), rat liver microsomes (RLM), and mouse liver microsomes (MLM) was investigated. DEET is metabolized by cytochromes P450 (P450s) leading to the production of a ring methyl oxidation product, N,N-diethyl m-hydroxymethylbenzamide (BALC), and an N-deethylated product, N-ethyl-m toluamide (ET). Both the affinities and intrinsic clearance of HLM for ring hydroxylation are greater than those for N-deethylation. Pooled HLM show significantly lower affinities (K(m)) than RLM for metabolism of DEET to either of the primary metabolites (BALC and ET). Among 15 cDNA-expressed P450 enzymes examined, CYP1A2, 2B6, 2D6*1 (Val(374)), and 2E1 metabolized DEET to the BALC metabolite, whereas CYP3A4, 3A5, 2A6, and 2C19 produced the ET metabolite. CYP2B6 is the principal cytochrome P450 involved in the metabolism of DEET to its major BALC metabolite, whereas CYP2C19 had the greatest activity for the formation of the ET metabolite. Use of phenotyped HLMs demonstrated that individuals with high levels of CYP2B6, 3A4, 2C19, and 2A6 have the greatest potential to metabolize DEET. Mice treated with DEET demonstrated induced levels of the CYP2B family, increased hydroxylation, and a 2.4-fold increase in the metabolism of chlorpyrifos to chlorpyrifos-oxon, a potent anticholinesterase. Preincubation of human CYP2B6 with chlorpyrifos completely inhibited the metabolism of DEET. Preincubation of human or rodent microsomes with chlorpyrifos, permethrin, and pyridostigmine bromide alone or in combination can lead to either stimulation or inhibition of DEET metabolism. PMID- 11854148 TI - N-Glucuronidation of some 4-arylalkyl-1H-imidazoles by rat, dog, and human liver microsomes. AB - N-Glucuronidation in vitro of six 4-arylalkyl-1H-imidazoles (both enantiomers of medetomidine, detomidine, atipamezole, and two other closely related compounds) by rat, dog, and human liver microsomes and by four expressed human UDP glucuronosyltransferase isoenzymes was studied. Human liver microsomes formed N glucuronides of 4-arylalkyl-1H-imidazoles with high activity, with apparent V(max) values ranging from 0.59 to 1.89 nmol/min/mg of protein. In comparison, apparent V(max) values for two model compounds forming the N-glucuronides 4 aminobiphenyl and amitriptyline were 5.07 and 0.56 nmol/min/mg of protein, respectively. Atipamezole showed an exceptionally low apparent K(m) value of 4.0 microM and a high specificity constant (V(max)/K(m)) of 256 compared with 4 aminobiphenyl (K(m), 265 microM; V(max)/K(m), 19) and amitriptyline (K(m), 728 microM; V(max)/K(m), 0.8). N-Glucuronidation of medetomidine was highly enantioselective in human liver microsomes; levomedetomidine exhibited a 60-fold V(max)/K(m) value compared with dexmedetomidine. Furthermore, two isomeric imidazole N-glucuronides were formed from dexmedetomidine, but only one was formed from levomedetomidine. Dog liver microsomes formed N-glucuronides of 4 arylalkyl-1H-imidazoles at a low rate and affinity, with apparent V(max) values ranging from 0.29 to 0.73 nmol/min/mg of protein and apparent K(m) values from 279 to 1640 microM. Rat liver microsomes glucuronidated these compounds at a barely detectable rate. Four expressed human UDP-glucuronosyltransferase isoenzymes (UGT1A3, UGT1A4, UGT1A6, and UGT1A9) were studied for 4-arylalkyl-1H imidazole-conjugating activity. Only UGT1A4 glucuronidated these compounds at an activity of about 5% of that measured for 4-aminobiphenyl. The observed activity of UGT1A4 does not explain the high efficiency of glucuronidation of 4-arylalkyl 1H-imidazoles in human liver microsomes. PMID- 11854149 TI - Phosphonate O-deethylation of [4-(4-bromo-2-cyano-phenylcarbamoyl) benzyl] phosphonic acid diethyl ester, a lipoprotein lipase-promoting agent, catalyzed by cytochrome P450 2C8 and 3A4 in human liver microsomes. AB - NO-1886 ([4-(4-bromo-2-cyano-phenylcarbamoyl) benzyl]-phosphonic acid diethyl ester) increases lipoprotein lipase activity, resulting in a reduction in plasma triglycerides and an increase in high-density lipoprotein cholesterol. The metabolism of NO-1886 in human liver was investigated in the present study. Ester cleavage of NO-1886 from diethyl phosphonate to monoethyl phosphonate was the major metabolic pathway catalyzed by cytochrome P450. In addition, the minor metabolic pathway in human liver was the hydrolysis of the amide bond of NO-1886 by a specific cytosolic esterase. Eadie-Hofstee plots of phosphonate O deethylation of NO-1886 in human liver microsomes showed a biphasic curve, indicating low- and high-K(m) components. Inhibition experiments with chemical inhibitors and antibodies against various cytochrome P450 isoforms suggested the involvement of CYP2C8 and CYP3A in the phosphonate O-deethylation. Recombinant CYP3A4 and CYP2C8 expressed in baculovirus-infected insect cells and human lymphoblastoid cells exhibited a high activity for phosphonate O-deethylation of NO-1886. The recombinant cytochrome P450 enzymes indicated that CYP2C8 and CYP3A4 were responsible for the low- and high-K(m) components in human liver microsomes, respectively. The selectivity of CYP2C8 in catalyzing phosphonate O-deethylation indicates that coadministration of drugs that are metabolized by the same enzyme requires careful consideration. PMID- 11854150 TI - Utility of rat liver slices to estimate hepatic clearance for application in physiologically based pharmacokinetic modeling: a study with tolbutamide, a compound with low extraction efficiency. AB - Liver slice experiments were performed to determine the slice intrinsic clearance and to extrapolate this to the in vivo liver intrinsic clearance in a physiologically based pharmacokinetic (PBPK)-like approach. Precision-cut liver slices were incubated with different initial concentrations of tolbutamide, and the time series of parent and metabolite concentrations were measured in slice and incubation medium. A mathematical model was built that modeled the uptake of tolbutamide and its metabolism in the liver slice. In addition, binding of tolbutamide to cellular constituents and partition over the water and lipid phase were accounted for by the model. Model analysis imposed sampling of parent compound in slice and of metabolites pooled from slice and medium. The model was calibrated to the data, fitting the intrinsic clearance, the parent compounds' free fraction in liver material, and a diffusion parameter describing medium slice exchange of tolbutamide. In addition, to ensure a meaningful application of the theoretical model, slice viability parameters were monitored before and during the experiment. For the different incubations, the intrinsic clearance per unit of volume of slice ranged from 0.035 to 0.086 min(-1) when not correcting for slice viability and from 0.044 to 0.11 min(-1) when correcting for slice viability. The results were extrapolated to a PBPK model for tolbutamide in the rat. The value for the intrinsic clearance found by calibrating the PBPK model to previous in vivo data was 0.090 min(-1). This result suggests that liver slices are a valuable tool for predicting in vivo intrinsic clearance of low-extraction compounds. PMID- 11854151 TI - Inhibition of cytochromes P450 by antifungal imidazole derivatives. AB - The interactions of a panel of antifungal agents with cytochromes P450 (P450s), as a means of predicting potential drug-drug interactions, have not yet been investigated. The objective of this study was to evaluate the specificity and selectivity of five antifungal agents using selective probe reactions for each of the eight major P450s. The index reactions used were phenacetin O-deethylation (for CYP1A2), coumarin 7-hydroxylation (CYP2A6), diclofenac 4'-hydroxylation (CYP2C9), omeprazole 5-hydroxylation (CYP2C19), dextromethorphan O-demethylation (CYP2D6), 7-ethoxy-4-trifluoromethylcoumarin deethylation (CYP2B6), chlorzoxazone 6-hydroxylation (CYP2E1), and omeprazole sulfonation (CYP3A4). Five antifungal agents that include an imidazole moiety (clotrimazole, miconazole, sulconazole, tioconazole, and ketoconazole) were examined in cDNA-expressing microsomes from human lymphoblast cells or human liver microsomes. All inhibitors studied demonstrated nonselective inhibition of P450s. Ketoconazole seemed to be the most selective for CYP3A4, although it also inhibited CYP2C9. High-affinity inhibition was seen for CYP1A2 (sulconazole and tioconazole K(i), 0.4 microM), CYP2B6 (miconazole K(i), 0.05 microM; sulconazole K(i), 0.04 microM), CYP2C19 (miconazole K(i), 0.05 microM; sulconazole K(i), 0.008 microM; tioconazole K(i), 0.04 microM), CYP2C9 (sulconazole K(i), 0.01 microM), CYP2D6 (miconazole K(i), 0.70 microM; sulconazole K(i), 0.40 microM), CYP2E1 (tioconazole K(i), 0.4 microM), and CYP3A4 (clotrimazole K(i), 0.02 microM; miconazole K(i), 0.03 microM; tioconazole K(i), 0.02 microM). Therefore, this class of compounds is likely to result in significant drug-drug interactions in vivo. PMID- 11854152 TI - Identification of the human cytochromes P450 responsible for atomoxetine metabolism. AB - Studies were performed to determine the human enzymes responsible for the biotransformation of atomoxetine to its major metabolite, 4-hydroxyatomoxetine, and to a minor metabolite, N-desmethylatomoxetine. Utilizing human liver microsomes containing a full complement of cytochrome P450 (P450) enzymes, average K(m) and CL(int) values of 2.3 microM and 103 microl/min/mg, respectively, were obtained for 4-hydroxyatomoxetine formation. Microsomal samples deficient in CYP2D6 exhibited average apparent K(m) and CL(int) values of 149 microM and 0.2 microl/min/mg, respectively. In a human liver bank characterized for P450 content, formation of 4-hydroxyatomoxetine correlated only to CYP2D6 activity. Of nine expressed P450s examined, 4-hydroxyatomoxetine was formed at a rate 475-fold greater by CYP2D6 compared with the other P450s. These results demonstrate that CYP2D6 is the enzyme primarily responsible for the formation of 4-hydroxyatomoxetine. Multiple P450s were found to be capable of forming 4-hydroxyatomoxetine when CYP2D6 was not expressed. However, the efficiency at which these enzymes perform this biotransformation is reduced compared with CYP2D6. The formation of the minor metabolite N desmethylatomoxetine exhibited average K(m) and CL(int) values of 83 microM and 0.8 microl/min/mg, respectively. Utilizing studies similar to those outlined above, CYP2C19 was identified as the primary enzyme responsible for the biotransformation of atomoxetine to N-desmethylatomoxetine. In summary, CYP2D6 was found to be the primary P450 responsible for the formation of the major oxidative metabolite of atomoxetine, 4-hydroxyatomoxetine. Furthermore, these studies indicate that in patients with compromised CYP2D6 activity, multiple low affinity enzymes will participate in the formation of 4-hydroxyatomoxetine. Therefore, coadministration of P450 inhibitors to poor metabolizers of CYP2D6 substrates would not be predicted to decrease the clearance of atomoxetine in these individuals. PMID- 11854153 TI - Glucuronidation of acetaminophen catalyzed by multiple rat phenol UDP glucuronosyltransferases. AB - Gunn rats glucuronidate acetaminophen (APAP) at reduced rates and show increased susceptibility to APAP-induced hepatotoxicity. This defect is presumed to involve UDP-glucuronosyltransferase (UGT) 1A6, which is nonfunctional in Gunn rats, but it is currently unclear whether other 1A family members are also involved. In humans, two 1A isoforms are known to be active (1A6 and 1A9) but 1A6 form has a 25-fold lower apparent K(m) (2 mM). Rat liver microsomal APAP UGT activity is induced by in vivo treatment with beta-naphthoflavone or oltipraz, an effect correlating with induction of 1A6 and 1A7. To address a possible role of 1A7 in APAP glucuronidation relative to other 1A forms, cDNAs encoding UGTs 1A1, 1A5, 1A6, 1A7, and 1A8 were expressed in human embryonic kidney cells and the contents of expressed enzyme in prepared membrane fractions determined by quantitative immunoblotting. At 2.5 mM APAP, 1A7 showed the highest specific activity (2.8 nmol/min/nmol 1A7 protein), followed by 1A6 (1.1 nmol/min/nmol), and 1A8 (0.27 nmol/min/nmol). 1A1 and 1A5 were essentially inactive. Kinetic comparisons indicated 1A7 had a similar apparent K(m) as 1A6 (4.7 versus 3.9 mM, respectively) but a 2.4-fold higher catalytic activity. These data suggest that in rats, 1A7 plays a major role in APAP glucuronidation and contributes to protection against APAP-induced hepatotoxicity. The involvement of other UGTs besides 1A6 is further underscored by the presence of significant residual APAP glucuronidating activity by Gunn rat hepatocytes, indicating the activity of an unknown UGT2 family member. PMID- 11854155 TI - The gastroprokinetic and antiemetic drug metoclopramide is a substrate and inhibitor of cytochrome P450 2D6. AB - Metoclopramide is increasingly prescribed for conditions previously treated with cisapride, but its metabolic enzymology and drug interactions are poorly understood. Using human liver microsomes (HLMs) and recombinant human cytochromes P450 (P450), we identified the major route of metoclopramide oxidation and the P450 isoforms involved. We also documented the ability of metoclopramide to inhibit the P450 system, using isoform-specific substrate reaction probes of CYP1A2, 2C19, 2C9, 2D6, 2E1, and 3A4. Metoclopramide was predominantly N dealkylated to monodeethylmetoclopramide, a metabolite that has not so far been described in humans. Formation rate of this metabolite followed Michaelis-Menten kinetics (K(m), 68 +/- 16 microM; V(max), 183 +/- 57 pmol/min/mg of protein; n = 3 HLMs). Of the isoform-specific inhibitors tested, 1 microM quinidine was a potent inhibitor of metoclopramide (25 microM) monodeethylation [by an average of 58.2%; range, approximately 38% (HL09-14-99) to 78.7% (HL161)] with K(i) values highly variable among the HLMs tested (K(i), mean +/- S.D., 2.7 +/- 2.8 microM; range, 0.15 microM in HL66, 2.4 microM in HL09-14-99, and 5.7 microM in HLD). Except troleandomycin, which inhibited metoclopramide metabolism in only one HLM (by approximately 23% in HL09-14-99), the effect of other inhibitors was minimal. Among the recombinant human P450 isoforms examined, monodeethylmetoclopramide was formed at the highest rate by CYP2D6 (V = 4.5 +/- 0.3 pmol/min/pmol of P450) and to a lesser extent by CYP1A2 (0.97 +/- 0.15 pmol/min/pmol of P450). The K(m) value derived (approximately 53 microM) was close to that from HLMs (68 microM). Metoclopramide is a potent inhibitor of CYP2D6 at therapeutically relevant concentrations (K(i) = 4.7 +/- 1.3 microM), with negligible effect on other isoforms tested. Further inhibition of CYP2D6 was observed when metoclopramide was preincubated with HLMs and NADPH-generating system before the substrate probe was added (maximum rate of inactivation, K(inact) = 0.02 min(-1), and the concentration required to achieve the half-maximal rate of inactivation, K'(i) = 0.96 microM), suggesting mechanism-based inhibition. Metoclopramide elimination is likely to be slowed in poor metabolizers of CYP2D6 or in patients taking inhibitors of this isoform, whereas metoclopramide itself could reduce the clearance of CYP2D6 substrate drugs. PMID- 11854154 TI - Glutathione and mercapturic acid conjugates of sulofenur and their activity against a human colon cancer cell line. AB - Sulofenur is one of the diarylsulfonylureas developed as an anticancer agent. Sulofenur possesses a broad spectrum of activity in several solid tumor models and has undergone extensive clinical trials based on its impressive preclinical activity. However, the clinical response of sulofenur has been disappointing because of the side effect of anemia. Furthermore, the anticancer mechanism of sulofenur and its diarylsulfonylurea analogs still remains unknown. Elucidation of the metabolic fates of sulofenur may help to delineate the mechanism and provide information to guide the structural modification for more potent anticancer agents with less side effects. We have identified a glutathione conjugate and a mercapturic acid conjugate from sulofenur-dosed rats with the aid of liquid chromatography/mass spectrometry. The fraction of the dose of sulofenur as the glutathione conjugate in the dosed-rat bile over 5 h was 0.12 +/- 0.03%, and the mercapturic acid conjugate in urine over 24 h was 1.4 +/- 0.7%. Protein binding of the glutathione conjugate and mercapturic acid conjugate was determined to be 20 +/- 3 and 84 +/- 2%, respectively, as opposed to >99% of sulofenur. The high protein binding of sulofenur requires a higher than in vitro dose, which is believed to cause the side effect of anemia. The significance of this metabolic pathway is that both conjugates were found to be glutathione reductase inhibitors and to possess anticancer activity comparable to sulofenur against human colon adenocarcinoma GC(3)/c1 cells, a sulofenur-sensitive cell line. These conjugates may serve as new leads for the development of novel anticancer agents. PMID- 11854156 TI - Chiral pharmacokinetics and inversion of enantiomers of a new quinoxaline topoisomerase IIbeta poison in the rat. AB - XK469 (NSC 697887; (+/-)-2-[4-(7-chloro-2-quinoxaliny)oxy]phenoxy propionic acid), an analog of the herbicide Assure, which possesses antitumor activity, especially against murine solid tumors and human xenografts, has recently been found to be the first topoisomerase IIbeta poison. Both R(+) and S(-) isomers are cytotoxic, although the R-isomer is more potent. Using a chiral high-performance liquid chromatography assay, pharmacokinetics of R(+)-, S(-)-, and (+/-)-XK469 in Fischer-344 rats were investigated following their separate i.v. administrations. S(-)-XK469 was found to be predominantly converted to the R-isomer in circulation when the S-isomer was administered either alone or as a racemic mixture. No trace of the S-isomer was found in circulation or in urine or feces, following the R isomer administration, up to 72 h. In the rat, the plasma concentration-time profiles for both isomers follow a two-compartment pharmacokinetics with the mean t(1/2beta) for the R-isomer of 24.7 h being significantly longer than 4.2 h, the mean t(1/2beta) for the S-isomer. The mean total clearance of the S-isomer was over 200-fold more rapid than that of the R-isomer, and the major clearance route of the S-enantiomer was inversion to its antipode, as estimated by the fractional formation clearance of R(+)-XK469 of 0.93. Protein binding for both enantiomers was in the range of 95 to 98%. Urinary and fecal elimination in 72 h as the intact drug were 7 to 10% and 8% of the administered dose, respectively, either administered as the individual enantiomers or as a racemate. Cumulative biliary elimination in 7 h was about 3% of the dose. No evidence of enantiomeric interaction at the pharmacokinetic level was detected. PMID- 11854157 TI - The alkaloid rutaecarpine is a selective inhibitor of cytochrome P450 1A in mouse and human liver microsomes. AB - Rutaecarpine, evodiamine, and dehydroevodiamine are quinazolinocarboline alkaloids isolated from a traditional Chinese medicine, Evodia rutaecarpa. The in vitro effects of these alkaloids on cytochrome P450 (P450)-catalyzed oxidations were studied using mouse and human liver microsomes. Among these alkaloids, rutaecarpine showed the most potent and selective inhibitory effect on CYP1A catalyzed 7-methoxyresorufin O-demethylation (MROD) and 7-ethoxyresorufin O deethylation (EROD) activities in untreated mouse liver microsomes. The IC(50) ratio of EROD to MROD was 6. For MROD activity, rutaecarpine was a noncompetitive inhibitor with a K(i) value of 39 +/- 2 nM. In contrast, rutaecarpine had no effects on benzo[a]pyrene hydroxylation (AHH), aniline hydroxylation, and nifedipine oxidation (NFO) activities. In human liver microsomes, 1 microM rutaecarpine caused 98, 91, and 77% decreases of EROD, MROD, and phenacetin O deethylation activities, respectively. In contrast, less than 15% inhibition of AHH, tolbutamide hydroxylation, chlorzoxazone hydroxylation, and NFO activities were observed in the presence of 1 microM rutaecarpine. To understand the selectivity of inhibition of CYP1A1 and CYP1A2, inhibitory effects of rutaecarpine were studied using liver microsomes of 3-methylcholanthrene (3-MC) treated mice and Escherichia coli membrane expressing bicistronic human CYP1A1 and CYP1A2. Similar to the CYP1A2 inhibitor furafylline, rutaecarpine preferentially inhibited MROD more than EROD and had no effect on AHH in 3-MC treated mouse liver microsomes. For bicistronic human P450s, the IC(50) value of rutaecarpine for EROD activity of CYP1A1 was 15 times higher than the value of CYP1A2. These results indicated that rutaecarpine was a potent inhibitor of CYP1A2 in both mouse and human liver microsomes. PMID- 11854167 TI - Biochemical, pathologic and behavioral analysis of a mouse model of glutaric acidemia type I. AB - Glutaric acidemia type I (GA-I) is an autosomal recessive disorder of amino acid metabolism resulting from a deficiency of glutaryl-CoA dehydrogenase (GCDH). Patients accumulate glutaric acid (GA) and 3-OH glutaric acid (3-OHGA) in their blood, urine and CSF. Clinically, GA-I is characterized by macrocephaly, progressive dystonia and dyskinesia. Degeneration of the caudate and putamen of the basal ganglia, widening of the Sylvian fissures, fronto-temporal atrophy and severe spongiform change in the white matter are also commonly observed. In this report we describe the phenotype of a mouse model of GA-I generated via targeted deletion of the Gcdh gene in embryonic stem cells. The Gcdh-/- mice have a biochemical phenotype very similar to human GA-I patients, including elevations of GA and 3-OHGA at levels similar to those seen in GA-I patients. The affected mice have a mild motor deficit but do not develop the progressive dystonia seen in human patients. Pathologically, the Gcdh-/- mice have a diffuse spongiform myelinopathy similar to that seen in GA-I patients. However, unlike in human patients, there is no evidence of neuron loss or astrogliosis in the striatum. Subjecting the Gcdh-/- mice to a metabolic stress, which often precipitates an encephalopathic crisis and the development of dystonia in GA-I patients, failed to have any neurologic effect on the mice. We hypothesize that the lack of similarity in regards to the neurologic phenotype and striatal pathology of GA-I patients, as compared with the Gcdh-/- mice, is due to intrinsic differences between the striata of mice and men. PMID- 11854168 TI - Twist haploinsufficiency in Saethre-Chotzen syndrome induces calvarial osteoblast apoptosis due to increased TNFalpha expression and caspase-2 activation. AB - Saethre-Chotzen syndrome (SCS) is a human autosomal dominant disorder characterized by premature fusion of cranial sutures caused by mutations of the Twist gene encoding a basic helix-loop-helix (bHLH) transcription factor. We previously showed that Twist haploinsufficiency caused by a Y103X nonsense mutation in SCS alters both proliferation and osteoblast gene expression in human calvarial osteoblasts, indicating that Twist is an important regulator of osteoblast differentiation. Here we show that Twist haploinsufficiency alters osteoblast apoptosis in SCS. Analysis of terminal deoxynucleotidyl transferase mediated nick-end labelling (TUNEL) demonstrated increased osteoblast and osteocyte apoptosis in coronal sutures from two SCS patients with nonsense mutations (Y103X and Q109X) that result in the synthesis of bHLH-truncated proteins, and one patient with a missense mutation in the basic domain (R118C) that abolishes Twist DNA binding. To assess the mechanisms involved, we studied osteoblast apoptosis in mutant (M-Tw) calvarial cells bearing the Y103X mutation resulting in decreased Twist mRNA and protein levels. M-Tw cells cultured in low serum conditions showed enhanced DNA fragmentation compared to normal (Nl) age matched calvarial cells. Biochemical analysis showed increased activity of initiator caspases-2 and -8 and downstream effector caspases-3, -6 and -7 in mutant osteoblasts. Caspase-2 was upstream of caspase-8 and effector caspases-3, 6 and -7 because their activities were suppressed by a specific caspase-2 inhibitor. M-Tw osteoblasts also showed increased cytochrome c release from the mitochondria. However, the activity of the downstream effector caspase-9 was not increased due to overexpression of the antagonist protein Hsp70. Detection of differentially expressed genes using cDNA expression array revealed increased Bax and TNFalpha mRNA levels in M-Tw compared to Nl cells, a finding confirmed by RT PCR and western blot analyses. Neutralization of TNFalpha overexpression using anti-TNFalpha or anti-TNF receptor 1 antibodies abolished the increased activity of caspase-2, caspase-8 and caspases-3, -6 and -7 in M-Tw osteoblasts. These studies provide novel evidence that Twist haploinsufficiency in SCS promotes osteoblast apoptosis by a TNFalpha-caspase-2-caspase-8-caspases-3, -6, -7 cascade, and uncover a molecular mechanism in which Twist plays an anti-apoptotic role in human calvarial osteoblasts. PMID- 11854169 TI - Premutation and intermediate-size FMR1 alleles in 10572 males from the general population: loss of an AGG interruption is a late event in the generation of fragile X syndrome alleles. AB - We previously reported a 1:259 prevalence of female carriers of FMR1 premutation size alleles (greater than 54 triplet repeats) in the general population. We now have screened 10 572 independent males from the same population for similar alleles using high-throughput Southern blotting. We identified 13 male carriers of an allele with more than 54 repeats. This corresponds to a prevalence of 1:813 males (95% confidence interval 1:527 to 1:1781). Haplotype analysis of four markers flanking the triplet array revealed that the prevalence of the major fragile X mutation-associated haplotype was increased among FMR1 alleles of 40-54 repeats. Although sequencing of highly unstable premutation alleles from fragile X families revealed only pure CGG tracts, this was not the case for alleles of similar size that were identified in males from the general population. Forty eight out of forty-nine alleles of 40 or more triplets had one or two AGG interruptions. This observation, combined with the observation of the enrichment of major fragile X syndrome haplotypes in all alleles of this size, is evidence that the loss of an AGG interruption in the triplet repeat array is not necessary for expansion of normal alleles of 29-30 triplets to intermediate size. The loss of AGG interruptions thus appears to be a late event that leads to greatly increased instability and may be related to the haplotype background of specific FMR1 alleles. PMID- 11854170 TI - Genotype/phenotype correlations of NPHS1 and NPHS2 mutations in nephrotic syndrome advocate a functional inter-relationship in glomerular filtration. AB - Mutations of the novel renal glomerular genes NPHS1 and NPHS2 encoding nephrin and podocin cause two types of severe nephrotic syndrome presenting in early life, Finnish type congenital nephrotic syndrome (CNF) and a form of autosomal recessive familial focal segmental glomerulosclerosis (SRN1), respectively. To investigate the mechanisms by which mutations might cause glomerular protein leak, we analysed NPHS1/NPHS2 genotype/phenotype relationships in 41 non-Finnish CNF patients, four patients with congenital (onset 0 to 3 months) focal segmental glomerulosclerosis and five patients with possible SRN1 (onset 6 months to 2 years). We clarify the range of NPHS1 mutations in CNF, detecting mutation 'hot spots' within the NPHS1 coding sequence. In addition, we describe a novel discordant CNF phenotype characterized by variable clinical severity, apparently influenced by gender. Moreover, we provide evidence that CNF may be genetically heterogeneous by detection of NPHS2 mutations in some CNF patients in whom NPHS1 mutations were not found. We confirm an overlap in the NPHS1/NPHS2 mutation spectrum with the characterization of a unique di-genic inheritance of NPHS1 and NPHS2 mutations, which results in a 'tri-allelic' hit and appears to modify the phenotype from CNF to one of congenital focal segmental glomerulosclerosis (FSGS). This may result from an epistatic gene interaction, and provides a rare example of multiple allelic hits being able to modify an autosomal recessive disease phenotype in humans. Our findings provide the first evidence for a functional inter-relationship between NPHS1 and NPHS2 in human nephrotic disease, thus underscoring their critical role in the regulation of glomerular filtration. PMID- 11854171 TI - KRIT1 association with the integrin-binding protein ICAP-1: a new direction in the elucidation of cerebral cavernous malformations (CCM1) pathogenesis. AB - Mutations in KRIT1, a protein initially identified based on a yeast two-hybrid interaction with the RAS-family GTPase RAP1A, are responsible for the development of the inherited vascular disorder cerebral cavernous malformations (CCM1). As the function of the KRIT1 protein and its role in CCM pathogenesis remain unknown, we performed yeast two-hybrid screens to identify additional protein binding partners. A fragment containing the N-terminal 272 amino acid residues of KRIT1, a region lacking similarity to any known protein upon database searches, was used as bait. From parallel screens of human fetal brain and HeLa cDNA libraries, we obtained multiple independent isolates of human integrin cytoplasmic domain-associated protein-1 (ICAP-1) as interacting clones. The interaction of KRIT1 and ICAP-1 was confirmed by GST-KRIT1 trapping of endogenous ICAP-1 from 293T cells. The alpha isoform of ICAP-1 is a 200 amino acid serine/threonine-rich phosphoprotein which binds the cytoplasmic tail of beta1 integrins. We show that mutagenesis of the N-terminal KRIT1 NPXY amino acid sequence, a motif critical for ICAP-1 binding to beta1 integrin molecules, completely abrogates the KRIT1/ICAP-1 interaction. The interaction between ICAP-1 and KRIT1, and the presence of a FERM domain in the latter, suggest that KRIT1 might be involved in the bidirectional signaling between integrin molecules and the cytoskeleton. Furthermore, these data suggest that KRIT1 might affect cell adhesion processes via integrin signaling in CCM1 pathogenesis. PMID- 11854172 TI - Aire deficient mice develop multiple features of APECED phenotype and show altered immune response. AB - Autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy (APECED) is a monogenic autosomal recessive disease caused by mutations in the AIRE gene. Here we have produced knock-out mice for the Aire gene. The Aire-/- mice develop normally; however, autoimmune features of APECED in Aire-/- mice are evident, including multiorgan lymphocytic infiltration, circulating autoantibodies and infertility. The distribution of B and T cells and thymic maturation as well as activation of T cells appear normal, while the TCR-Vbeta repertoire is altered in peripheral T cells of Aire-/- mice. When mice are challenged with immunization, the peripheral T cells of Aire-/- mice have a 3-5-fold increased proliferation. These findings suggest that the Aire gene is not necessary for normal T cell education and development, while a defect in immune response detected in challenged Aire-/- mice underlines the crucial role of AIRE/Aire in maintaining homeostatic regulation in the immune system. PMID- 11854173 TI - A human H19 transgene exhibits impaired paternal-specific imprint acquisition and maintenance in mice. AB - Genomic imprinting, the differential expression of autosomal genes based on their parent of origin, is observed in all eutherian mammals that have been examined. In most instances the genes that are imprinted in one species are imprinted in others as well, suggesting that imprinting predated eutherian radiation. For example, the RNA-coding H19 gene is repressed upon paternal inheritance in all species examined to date. Thus, it is surprising that there is remarkably little sequence conservation among the cis-acting DNA regulatory elements that are required for imprinting of H19 and the tightly linked Igf2 gene. The most conserved characteristic in the imprinting control region (ICR) is the presence of multiple binding sites for the zinc finger protein CTCF, raising the possibility that CTCF binding might be sufficient for the reciprocal imprinting of H19 and Igf2. To investigate whether a human H19 transgene, harboring seven CTCF sites, is correctly recognized and imprinted in the mouse, a 100 kb transgene containing the human H19 gene was introduced into the mouse germline. The human transgene was specifically methylated after passage through the male germline in a copy number-dependent manner, but the methylation was unstable, undergoing progressive loss during development. Consequently, the transgene was highly expressed upon both maternal and paternal inheritance. These results argue that the signals for both the acquisition and maintenance of methylation imprinting are diverging rapidly. PMID- 11854174 TI - Heterogeneity of linkage disequilibrium in human genes has implications for association studies of common diseases. AB - Linkage disequilibrium (LD) is the central concept of genetic association studies. Although LD has been shown not to be uniformly distributed across the genome, limited information is available about the characteristics of LD within candidate genes at large. We screened coding and regulatory regions of 50 candidate genes for cardiovascular diseases and identified 228 polymorphisms. The overall sequence diversity was 3.81 +/- 0.31 x 10(-4). Intragenic LD was generally very strong, with 40% of the 464 pairs of polymorphisms exhibiting a complete LD. However, if we consider /D'/ = 0.7 as an arbitrary limit for useful LD in association studies, 26% of the pairs fell below this threshold, half of which being in negative LD, a situation where LD is even more difficult to detect. Non-synonymous coding polymorphisms, which are more likely to have a functional role, were more represented among low-frequency alleles and were more often in complete negative LD with other polymorphisms. This implies that in many situations the power to detect the effect of a non-synonymous polymorphism by measuring a nearby marker might be low. Although intragenic LD was partly a function of physical distance, gene-specific patterns of LD were observed, making it difficult to provide general guidelines for selecting the most useful polymorphisms in association studies. For all these reasons, association studies should concentrate on the overall sequence variation of functionally important regions of candidate genes and not only on a few polymorphisms. The variability of important intergenic regions identified by different approaches including comparative genomics will also have to be assessed. PMID- 11854176 TI - Breaks at telomeres and TRF2-independent end fusions in Fanconi anemia. AB - Fanconi anemia (FA) is a rare genetic disease characterized by chromosome instability, progressive pancytopenia and cancer susceptibility. Telomeres are intimately related to chromosome stability and play an important role in organismal viability at the hematological level. Since previous works suggested an accelerated shortening of telomeres in FA, we have studied several markers of telomere integrity and function in FA patients and age-matched controls to get insights into the mechanisms and consequences of telomere erosion in FA. A higher frequency of extra-chromosomic TTAGGG signals and of chromosome ends with undetectable TTAGGG repeats was observed in FA cells by fluorescence in situ hybridization (FISH), suggesting intensive breakage at telomeric sequences. This was proven by measuring the frequency of excess of telomeric signals per cell, which was 2.8-fold higher in FA. Consistent with previous reports, quantitative FISH analysis showed an accelerated telomere shortening of 0.68 kb in FA, which occurred concurrently in both chromosome arms in a similar magnitude. Our data therefore suggest that the telomere erosion in FA is caused by a higher rate of breakage at TTAGGG sequences in vivo in differentiated cells, in addition to mere replicative shortening during lymphocyte proliferation. Consistent with impaired telomeres in FA patients, we observed a >10-fold increase in chromosome end fusions in FA compared to normal controls. This observation was independent of TRF2, a telomere binding factor that protects human telomeres from end fusions, since immunohistochemistry studies in FA cell lines and corrected counterparts by retrovirus-mediated transfer of FANCA and FANCD2 cDNA showed that a functional FA pathway is not required for telomere binding of TRF2. PMID- 11854175 TI - Differentiation-specific effects of LHON mutations introduced into neuronal NT2 cells. AB - Inheritance of one of three primary mutations at positions 11778, 3460 or 14484 of the mitochondrial genome in subunits of Complex I causes Leber's Hereditary Optic Neuropathy (LHON), a specific degeneration of the optic nerve, resulting in bilateral blindness. It has been unclear why inheritance of a systemic mitochondrial mutation would result in a specific neurodegeneration. To address the neuron-specific degenerative phenotype of the LHON genotype, we have created cybrids using a neuronal precursor cell line, Ntera 2/D1 (NT2), containing mitochondria from patient lymphoblasts bearing the most common LHON mutation (11778) and the most severe LHON mutation (3460). The undifferentiated LHON-NT2 mutant cells were not significantly different from the parental cell control in terms of mtDNA/nDNA ratio, mitochondrial membrane potential, reactive oxygen species (ROS) production or the ability to reduce Alamar Blue. Differentiation of NT2s resulted in a neuronal morphology and neuron-specific pattern of gene expression, and a 3-fold reduction in mtDNA/nDNA ratio in both mutant and control cells; however, the differentiation protocol yielded significantly less LHON cells than controls, by 30%, indicating either a decreased proliferative potential or increased cell death of the LHON-NT2 cells. Differentiation of the cells to the neuronal form also resulted in significant increases in ROS production in the LHON-NT2 neurons versus controls, which is abolished by rotenone, a specific inhibitor of Complex I. We infer that the LHON genotype requires a differentiated neuronal environment in order to induce increased mitochondrial ROS, which may be the cause of the reduced NT2 yield; and suggest that the LHON degenerative phenotype may be the result of an increase in mitochondrial superoxide which is caused by the LHON mutations, possibly mediated through neuron-specific alterations in Complex I structure. PMID- 11854177 TI - Distinct PTEN mutational spectra in hereditary non-polyposis colon cancer syndrome-related endometrial carcinomas compared to sporadic microsatellite unstable tumors. AB - Germline PTEN mutations cause Cowden syndrome (CS) and Bannayan-Riley-Ruvalcaba syndrome (BRR), two hamartoma-tumor syndromes with an increased risk of breast, thyroid and endometrial cancers. Somatic genetic and epigenetic inactivation of PTEN is involved in as high as 93% of sporadic endometrial carcinomas (EC), irrespective of microsatellite status, and can occur in the earliest precancers. EC is the most frequent extra-colonic cancer in patients with hereditary non polyposis colon cancer syndrome (HNPCC), characterized by germline mutations in the mismatch repair (MMR) genes and by microsatellite instability (MSI) in component tumors. To determine whether PTEN is involved in the pathogenesis of EC arising in HNPCC cases, and whether PTEN inactivation precedes MMR deficiency, we obtained 41 ECs from 29 MLH1 or MSH2 mutation positive HNPCC families and subjected them to PTEN expression and mutation analysis. Immunohistochemical analysis revealed 68% (28/41) of the HNPCC-related ECs with absent or weak PTEN expression. The remaining 27% (11/41) of tumors had normal expression and 5% (2/41) with mixed populations showing weak/absent as well as normal expression. Mutation analysis of 20 aberrant PTEN-expressing tumors revealed that 17 (85%) harbored 18 somatic PTEN mutations. All mutations were frameshift, 10 (56%) of which involved the 6(A) tracts in exon 7 or 8. These results suggest that PTEN plays a significant pathogenic role in both HNPCC and sporadic endometrial carcinogenesis, unlike the scenarios for colorectal cancer. Furthermore, we have shown that somatic PTEN mutation, especially frameshift, is a consequence of profound MMR deficiency in HNPCC-related ECs. In contrast, among 60 previously reported MSI+ sporadic ECs with 70 somatic mutations in PTEN, 39 (56%) were frameshift, of which only eight (21%) were affecting the 6(A) tracts in exon 7 or 8 (P = 0.01), suggesting that PTEN mutations may precede MMR deficiency. PMID- 11854178 TI - Categorization and characterization of transcript-confirmed constitutively and alternatively spliced introns and exons from human. AB - By spliced alignment of human DNA and transcript sequence data we constructed a data set of transcript-confirmed exons and introns from 2793 genes, 796 of which (28%) were seen to have multiple isoforms. We find that over one-third of human exons can translate in more than one frame, and that this is highly correlated with G+C content. Introns containing adenosine at donor site position +3 (A3), rather than guanosine (G3), are more common in low G+C regions, while the converse is true in high G+C regions. These two classes of introns are shown to have distinct lengths, consensus sequences and correlations among splice signals, leading to the hypothesis that A3 donor sites are associated with exon definition, and G3 donor sites with intron definition. Minor classes of introns, including GC-AG, U12-type GT-AG, weak, and putative AG-dependant introns are identified and characterized. Cassette exons are more prevalent in low G+C regions, while exon isoforms are more prevalent in high G+C regions. Cassette exon events outnumber other alternative events, while exon isoform events involve truncation twice as often as extension, and occur at acceptor sites twice as often as at donor sites. Alternative splicing is usually associated with weak splice signals, and in a majority of cases, preserves the coding frame. The reported characteristics of constitutive and alternative splice signals, and the hypotheses offered regarding alternative splicing and genome organization, have important implications for experimental research into RNA processing. The 'AltExtron' data sets are available at http://www.bit.uq.edu.au/altExtron/ and http://www.ebi.ac.uk/~thanaraj/altExtron/. PMID- 11854179 TI - A candidate molecular mechanism for the association of an intronic polymorphism of FE65 with resistance to very late onset dementia of the Alzheimer type. AB - Late onset dementias of the Alzheimer type may be coupled to intrinsic aging processes. Their major pathological hallmarks are the deposition of aggregates of beta amyloid (Abeta) peptides, proteolytic products from internal portions of the Abeta precursor protein, betaPP. Susceptibility appears to be modulated by polymorphic alleles at multiple loci. Most of these putative assignments, however, have been controversial. It is therefore essential to provide evidence of a plausible biological basis for each such association. Here, we show such evidence for the case of a biallelic polymorphism of the FE65 intron 13. FE65 is an adaptor protein that tightly binds to the cytoplasmic tail of betaPP. Increasing evidence indicates that this binding plays a critical role in a signaling pathway. Our results reveal that a protective (minor) allele alters the splicing of the terminal exon by selection of an alternative acceptor site, resulting in an isoform, FE65a2, with an altered C-terminal region lacking part of a betaPP binding site. Pull down assays confirmed that the FE65a2 isoform binds to betaPP less efficiently, suggesting that an attenuated binding of FE65 with betaPP is, in part, responsible for resistance to the very late onset disease. Sequence analysis of the FE65 of mice, non-human primates and man revealed that the susceptibility allele, which codes for strong binding of the FE65 protein with betaPP, was favored by natural selection leading to our lineage. That allele may contribute to very late onset form of Alzheimer disease when we are aged. PMID- 11854181 TI - Penicillium marneffei causes osteopontin-mediated production of interleukin-12 by peripheral blood mononuclear cells. AB - We investigated the role of osteopontin (OPN) in interleukin-12 (IL-12) production from peripheral blood mononuclear cells (PBMCs) stimulated with Penicillium marneffei. Kinetic studies showed that OPN synthesis preceded that of IL-12 at both mRNA and protein levels when PBMCs were cocultured with P. marneffei. Treatment with anti-OPN monoclonal antibodies (MAb) significantly suppressed IL-12 secretion. Furthermore, native OPN induced a profound level of synthesis of IL-12 from noninfected PBMCs. The major cellular source of OPN was monocytes, because depletion of CD14(+) cells resulted in the abrogation of such production. We also examined the regulatory role of granulocyte-macrophage colony stimulating factor (GM-CSF) in OPN secretion from P. marneffei-stimulated PBMCs. Neutralizing anti-GM-CSF MAb significantly reduced OPN secretion, and treatment with this cytokine induced OPN production from both infected and noninfected PBMCs. Finally, antagonists against the mannose receptor but not the beta-glucan receptor almost completely abrogated the production of OPN. Our results demonstrated that OPN secreted from monocytes is involved in the production of IL 12 from PBMCs after stimulation with P. marneffei and that OPN production is regulated by GM-CSF. Our results also indicated the possible involvement of the mannose receptor as a signal-transducing receptor for triggering the secretion of OPN by P. marneffei-stimulated PBMCs. PMID- 11854180 TI - Toll receptors: a central element in innate immune responses. PMID- 11854182 TI - Essential role of interleukin-12 (IL-12) and IL-18 for gamma interferon production induced by listeriolysin O in mouse spleen cells. AB - The mechanism of gamma interferon (IFN-gamma) production induced by listeriolysin O (LLO), a cytolytic virulence factor of Listeria monocytogenes, was analyzed with special reference to the involvement of macrophage-derived cytokines in spleen cells of mice. LLO purified from the culture supernatant of L. monocytogenes was capable of inducing a high level of IFN-gamma when its cytolytic activity was blocked by cholesterol treatment. The IFN-gamma-inducing ability of LLO was not dependent on possibly contaminating lipopolysaccharide. Depletion of CD11b(+) cells resulted in a profound decrease in IFN-gamma production in response to LLO stimulation. Negative selection also suggested the contribution of DX5(+) cells in IFN-gamma production. Reverse transcription-PCR revealed that expression of interleukin-12 (IL-12) p35 and p40 was induced by LLO but that the IL-18 mRNA level in the CD11b(+) fraction of spleen cells was unchanged. There was no change in the expression of the IFN-gamma-inducing cytokine genes in the CD11b(-) fraction. Neutralization of IL-12 and IL-18 in culture abolished the IFN-gamma production almost completely. Spleen cells from IL-12- or IL-18-deficient mice never produced IFN-gamma after stimulation with LLO. These results clearly indicated that LLO, a well-known virulence factor of L. monocytogenes, is capable of inducing IFN-gamma from NK cells through induction of IL-12 and IL-18 from macrophages. LLO appeared to play essential roles, not only as a bacterial virulence factor but also as a bacterial modulin in the immune response of the host. PMID- 11854183 TI - Transcutaneous immunization using colonization factor and heat-labile enterotoxin induces correlates of protective immunity for enterotoxigenic Escherichia coli. AB - Enterotoxigenic Escherichia coli (ETEC) diarrheal disease is a worldwide problem that may be addressed by transcutaneous delivery of a vaccine. In several human settings, protective immunity has been associated with immune responses to E. coli colonization factors and to the heat-labile toxin that induces the diarrhea. In this set of animal studies, transcutaneous immunization (TCI) using recombinant colonization factor CS6 and cholera toxin (CT) or heat-labile enterotoxin (LT) as the adjuvant induced immunoglobulin G (IgG) and IgA anti-CS6 responses in sera and stools and antibody responses that recognized CS6 antigen in its native configuration. The antitoxin immunity induced by TCI was also shown to protect against enteric toxin challenge. Although immunization with LT via the skin induced mucosal secretory IgA responses to LT, protection could also be achieved by intravenous injection of the immune sera. Finally, a malaria vaccine antigen, merzoite surface protein 1(42) administered with CT as the adjuvant, induced both merzoite surface protein antibodies and T-cell responses while conferring protective antitoxin immunity, suggesting that both antiparasitic activity and antidiarrheal activity can be obtained with a single vaccine formulation. Overall, our results demonstrate that relevant colonization factor and antitoxin immunity can be induced by TCI and suggest that an ETEC traveler's diarrhea vaccine could be delivered by using a patch. PMID- 11854184 TI - Passive intranasal monoclonal antibody prophylaxis against murine Pneumocystis carinii pneumonia. AB - Passive antibody immunoprophylaxis is one method used to protect patients against infection if they are unable to mount an adequate active immune response. Topical application of antibody may be effective against infections at mucosal sites. Using a SCID mouse model of Pneumocystis carinii pneumonia, we were able to demonstrate protection against an airborne challenge with P. carinii by intranasal administration of antibody. Immunoglobulin M (IgM) monoclonal antibodies to an epitope shared by mouse and human P. carinii organisms reduced organism numbers by more than 99% under the conditions described. An IgG1 switch variant of one of the IgM monoclonal antibodies was also protective. These experiments provide a model for exploring the utility of this approach in protecting at-risk patients from infection with P. carinii. PMID- 11854185 TI - Role of lung epithelial cells in defense against Klebsiella pneumoniae pneumonia. AB - The airway epithelium represents a primary site for the entry of pathogenic bacteria into the lungs. It has been suggested for many respiratory pathogens, including Klebsiella pneumoniae, that adhesion and invasion of the lung epithelial cells is an early stage of the pneumonia process. We observed that poorly encapsulated K. pneumoniae clinical isolates and an isogenic unencapsulated mutant invaded lung epithelial cells more efficiently than highly encapsulated strains independent of the K type. By contrast, the unencapsulated mutant was completely avirulent in a mouse model of pneumonia, unlike the wild type strain, which produced pneumonia and systemic infection. Furthermore, the unencapsulated mutant bound more epithelially produced complement component C3 than the wild-type strain. Our results show that lung epithelial cells play a key role as a host defense mechanism against K. pneumoniae pneumonia, using two different strategies: (i) ingestion and control of the microorganisms and (ii) opsonization of the microorganisms. Capsular polysaccharide avoids both mechanisms and enhances the virulence of K. pneumoniae. PMID- 11854186 TI - Correlation between an in vitro invasion assay and a murine model of Burkholderia cepacia lung infection. AB - Our understanding of the virulence of Burkholderia cepacia complex lung infections in cystic fibrosis patients is incomplete. There is a great deal of variability in the clinical course, from simple colonization to severe and often fatal necrotizing pneumonia, termed cepacia syndrome. Multiple subspecies (called genomovars) have been identified, and these genomovars may hold the key to understanding the variable pathogenicity. Thirty-one B. cepacia complex isolates belonging to five of the seven genomovars were examined by using a gentamicin protection assay of invasion with A549 cells. The level of epithelial cell invasion by B. cepacia in the A549 model was relatively low compared with the data obtained for other pathogens and was often variable from assay to assay. Thus, a statistical approach was used to determine invasiveness. When this model was used, one of four genomovar I strains (25%), three of eight genomovar II strains (37.5%), seven of nine genomovar III strains (77.8%), one of four genomovar IV strains (25%), and none of the four genomovar V strains examined were defined as invasive. All other strains were categorized as either noninvasive or indeterminate. Invasive, noninvasive, and indeterminate isolates belonging to genomovars II and III were subsequently tested for splenic invasion with the mouse agar bead model. Correlation between the models for six strains was demonstrated. Our results indicate that a statistical model used to determine invasiveness in an in vitro invasion assay can be used to predict in vivo invasiveness. PMID- 11854187 TI - Intracellular induction of Listeria monocytogenes actA expression. AB - Following entry into the host cytosol, the bacterial pathogen Listeria monocytogenes dramatically increases the expression of several key virulence factors. The expression of actA, whose protein product is required for L. monocytogenes actin-based intracellular motility, is increased by more than 200 fold in cytosolic bacteria in comparison to broth-grown cultures. Two distinct promoter elements have been reported to regulate actA expression. One promoter is located immediately upstream of actA coding sequences, while the second promoter is contributed by the upstream mpl gene via the generation of an mpl-actA-plcB transcript. A series of L. monocytogenes mutants were constructed to define the contributions of individual promoter elements to actA expression. The intracellular induction of actA expression was found to be dependent upon the actA proximal promoter; the mpl promoter appeared to contribute to the extracellular induction of actA but did not affect intracellular levels of expression. The actA promoter is dependent upon a regulatory factor known as PrfA for transcriptional activation; however, no increase in actA expression was detected following the introduction of a high-affinity PrfA binding site within the actA promoter. The presence of a mutationally activated form of PrfA, known as PrfA*, increased overall actA expression in broth-grown cultures of both wild type and actA promoter mutant strains, but the levels of induction observed were still approximately 50-fold lower than those observed for intracellularly grown L. monocytogenes. Collectively, these results indicate that the dramatic induction of actA expression that occurs in the host cell cytosol is mediated through a single promoter element. Furthermore, intracellular induction of actA appears to require additional steps or factors beyond those necessary for the activation and binding of PrfA to the actA promoter. PMID- 11854188 TI - Dendritic cells pulsed with a recombinant chlamydial major outer membrane protein antigen elicit a CD4(+) type 2 rather than type 1 immune response that is not protective. AB - Chlamydia trachomatis is an obligate intracellular bacterium that infects the oculogenital mucosae. C. trachomatis infection of the eye causes trachoma, the leading cause of preventable blindness. Infections of the genital mucosae are a leading cause of sexually transmitted diseases. A vaccine to prevent chlamydial infection is needed but has proven difficult to produce by using conventional vaccination approaches. Potent immunity to vaginal rechallenge in a murine model of chlamydial genital infection has been achieved only by infection or by immunization with dendritic cells (DC) pulsed ex vivo with whole inactivated organisms. Immunity generated by infection or ex vivo antigen-pulsed DC correlates with a chlamydia-specific interleukin 12 (IL-12)-dependent CD4(+) Th1 immune response. Because of the potent antichlamydial immunizing properties of DC, we hypothesized that DC could be a powerful vehicle for the delivery of individual chlamydial antigens that are thought to be targets for more conventional vaccine approaches. Here, we investigated the recombinant chlamydial major outer membrane protein (rMOMP) as a target antigen. The results demonstrate that DC pulsed with rMOMP secrete IL-12 and stimulate infection-sensitized CD4(+) T cells to proliferate and secrete gamma interferon. These immunological properties implied that rMOMP-pulsed DC would be potent inducers of MOMP-specific CD4(+) Th1 immunity in vivo; however, we observed the opposite result. DC pulsed ex vivo with rMOMP and adoptively transferred to naive mice generated a Th2 rather than a Th1 anti-MOMP immune response, and immunized mice were not protected following infectious challenge. We conclude from these studies that the immunological properties of ex vivo pulsed DC are not necessarily predictive of the immune response generated in vivo following adoptive transfer. These findings suggest that the nature of the antigen used to pulse DC ex vivo influences the Th1-Th2 balance of the immune response in vivo. PMID- 11854189 TI - Identification of a second Arcanobacterium pyogenes neuraminidase and involvement of neuraminidase activity in host cell adhesion. AB - Arcanobacterium pyogenes, a common inhabitant of the upper respiratory and urogenital tracts of economically important animals, such as cattle and swine, is also an opportunistic pathogen associated with suppurative infections in these animals. A. pyogenes expresses neuraminidase activity encoded by the nanH gene, and previously, construction of a nanH mutant of A. pyogenes BBR1 indicated that a second neuraminidase is present in this strain. A 5,112-bp gene, nanP, was cloned and sequenced, and this gene conferred neuraminidase activity on an Escherichia coli host strain. The predicted 186.8-kDa NanP protein exhibited similarity to a number of bacterial neuraminidases and contained the RIP/RLP motif and five copies of the Asp box motif found in all bacterial neuraminidases. As expected, insertional inactivation of the nanP gene in A. pyogenes BBR1 resulted in a mutant with reduced neuraminidase activity. However, insertional inactivation of the nanP gene in the nanH mutant strain resulted in a complete lack of neuraminidase activity. Like NanH, NanP was localized to the A. pyogenes cell wall. However, unlike the nanH gene, which was present in 100% of the strains examined, nanP was present in only 64.2% of the isolates (n = 53). A. pyogenes adheres to HeLa cells, and a nanP mutant displayed a wild-type adhesion phenotype with these cells. In contrast, the ability of a nanH nanP double mutant to bind to HeLa cells was reduced by 53%. The wild-type adhesion phenotype was restored by providing nanP in trans. These data indicate that the neuraminidases of A. pyogenes play a role in adhesion of this organism to host epithelial cells. PMID- 11854190 TI - Role of glutathione metabolism of Treponema denticola in bacterial growth and virulence expression. AB - Hydrogen sulfide (H(2)S) is a major metabolic end product detected in deep periodontal pockets that is produced by resident periodontopathic microbiota associated with the progression of periodontitis. Treponema denticola, a member of the subgingival biofilm at disease sites, produces cystalysin, an enzyme that catabolizes cysteine, releasing H(2)S. The metabolic pathway leading to H(2)S formation in periodontal pockets has not been determined. We used a variety of thiol compounds as substrates for T. denticola to produce H(2)S. Our results indicate that glutathione, a readily available thiol source in periodontal pockets, is a suitable substrate for H(2)S production by this microorganism. In addition to H(2)S, glutamate, glycine, ammonia, and pyruvate were metabolic end products of metabolism of glutathione. Cysteinyl glycine (Cys-Gly) was also catabolized by the bacteria, yielding glycine, H(2)S, ammonia, and pyruvate. However, purified cystalysin could not catalyze glutathione and Cys-Gly degradation in vitro. Moreover, the enzymatic activity(ies) in T. denticola responsible for glutathione breakdown was inactivated by trypsin or proteinase K, by heating (56 degrees C) and freezing (-20 degrees C), by sonication, and by exposure to N alpha-p-tosyl-L-lysine chloromethyl ketone (TLCK). These treatments had no effect on degradation of cysteine by the purified enzyme. In this study we delineated an enzymatic pathway for glutathione metabolism in the oral spirochete T. denticola; our results suggest that glutathione metabolism plays a role in bacterial nutrition and potential virulence expression. PMID- 11854191 TI - Characterization of a cytotoxic factor in culture filtrates of Serratia marcescens. AB - Serratia marcescens culture filtrates have been reported to be cytotoxic to mammalian cells. Using biochemical and genetic approaches, we have identified a major source of this cytotoxic activity. Both heat and protease treatments abrogated the cytotoxicity of S. marcescens culture filtrates towards HeLa cells, suggesting the involvement of one or more protein factors. A screen for in vitro cytotoxic activity revealed that S. marcescens mutant strains that are deficient in production of a 56-kDa metalloprotease are significantly less cytotoxic to mammalian cells. Cytotoxicity was significantly reduced when culture filtrates prepared from wild-type strains were pretreated with either EDTA or 1,10 phenanthroline, which are potent inhibitors of the 56-kDa metalloprotease. Furthermore, cytotoxic activity was restored when the same culture filtrates were incubated with zinc divalent cations, which are essential for enzymatic activity of the 56-kDa metalloprotease. Finally, recombinant expression of the S. marcescens 56-kDa metalloprotease conferred a cytotoxic phenotype on the culture filtrates of a nonpathogenic Escherichia coli strain. Collectively, these data suggest that the 56-kDa metalloprotease contributes significantly to the in vitro cytotoxic activity commonly observed in S. marcescens culture filtrates. PMID- 11854192 TI - Site-specific proteolysis of the MALP-404 lipoprotein determines the release of a soluble selective lipoprotein-associated motif-containing fragment and alteration of the surface phenotype of Mycoplasma fermentans. AB - The mature MALP-404 surface lipoprotein of Mycoplasma fermentans comprises a membrane-anchored N-terminal lipid-modified region responsible for macrophage activation (P. F. Muhlradt, M. Kiess, H. Meyer, R. Sussmuth, and G. Jung, J. Exp. Med. 185:1951-1958, 1997) and an external hydrophilic region that contains the selective lipoprotein-associated (SLA) motif defining a family of lipoproteins from diverse but selective prokaryotes, including mycoplasmas (M. J. Calcutt, M. F. Kim, A. B. Karpas, P. F. Muhlradt, and K. S. Wise, Infect. Immun. 67:760-771, 1999). This family generally corresponds to a computationally defined group of orthologs containing the basic membrane protein (BMP) domain. Two discrete lipid modified forms of the abundant MALP product which vary dramatically in ratio among isolates of M. fermentans occur on the mycoplasma surface: (i) MALP-404, the full-length mature product, and (ii) MALP-2, the Toll-like receptor 2 mediated macrophage-activating lipopeptide containing the N-terminal 14 residues of the mature lipoprotein. The role of posttranslational processing in the biogenesis of MALP-2 from the prototype MALP-404 SLA-containing lipoprotein was investigated. Detergent phase fractionation of cell-bound products and N-terminal sequencing of a newly discovered released fragment (RF) demonstrated that MALP 404 was subject to site-specific proteolysis between residues 14 and 15 of the mature lipoprotein, resulting in the cell-bound MALP-2 and soluble RF products. This previously unknown mechanism of posttranslational processing among mycoplasmas suggests that specific cleavage of some surface proteins may confer efficient "secretion" of extracellular products by these organisms, with concurrent changes in the surface phenotype. This newly identified form of variation may have significant implications for host adaptation by mycoplasmas, as well as other pathogens expressing lipoproteins of the SLA (BMP) family. PMID- 11854193 TI - Identification and characterization of a nonimmunoglobulin factor in human saliva that inhibits Streptococcus mutans glucosyltransferase. AB - Saliva contains an array of nonimmunoglobulin defense factors which are thought to contribute to the protection of the hard and soft tissue surfaces of the oral cavity by modulating microbial colonization and metabolism. Here we report the discovery of a putative innate defense factor in human saliva that inhibits the glucosyltransferase (GTF) of Streptococcus mutans, a virulence enzyme involved in oral colonization by this pathogen. The GTF-inhibiting factor (GIF) was initially identified as a nonimmunoglobulin salivary component that interfered with detection of antibodies to the glucan-binding region (GLU) of GTF by an enzyme linked immunosorbent assay. This inhibitory activity was present in whole saliva and submandibular-sublingual saliva, but it was essentially absent from parotid saliva. GIF inhibited the recognition of S. mutans cell surface-associated GTF by specific antibodies but had no effect on antibodies to other cell surface antigens, suggesting that GIF specifically binds to GTF on S. mutans. GIF purified by size exclusion or affinity chromatography was used for biochemical and functional characterization. Analysis of GIF by sodium dodecyl sulfate polyacrylamide gel electrophoresis revealed a high-molecular-weight glycoprotein after staining with Coomassie blue or Schiff's reagent. Heating and reduction with 2-mercaptoethanol of GIF resulted in the release of a approximately 58-kDa protein that was identified as alpha-amylase by Western blotting using anti-alpha amylase antibodies. GLU bound blotted alpha-amylase, suggesting that the latter molecule is the GLU-binding component of the GIF complex. The ability of GTF to synthesize extracellular glucans was inhibited by GIF but not by uncomplexed alpha-amylase or an unrelated high-molecular-weight glycoprotein. In conclusion, our findings demonstrate that in human saliva, there is a high-molecular-weight glycoprotein-alpha-amylase complex which is capable of inhibiting GTF and may contribute to control of S. mutans colonization in the oral cavity. PMID- 11854194 TI - Cross-protective immunity of mice induced by oral immunization with pneumococcal surface adhesin a encapsulated in microspheres. AB - The global use of a capsular polysaccharide-based pneumococcal vaccine has been limited because of serotype-specific protection and poor effectiveness in individuals with low immunocompetency. The mucosal immune system develops earlier in infants and lasts longer in the elderly than does the systemic immune system. Furthermore, mucosal immunization is beneficial for AIDS patients, because human immunodeficiency virus-infected subjects can develop normal mucosal antibody responses even in late clinical phases. For these reasons, we evaluated recombinant pneumococcal surface adhesin A (rPsaA) of Streptococcus pneumoniae in terms of cross-protective immune responses after oral delivery. Encapsulated rPsaA provided higher immunogenicity than naked rPsaA. Coencapsulation or codelivery of the cholera toxin (CT) B subunit (CTB) and CT also increased the immunogenicity of rPsaA. Cross-protective immunities against five strains of S. pneumoniae (types 4, 6B, 14, 19F, and 23F) were induced after oral immunization with microencapsulated rPsaA. Lung colonization and septicemia caused by the five serotypes were significantly inhibited by oral immunization with microencapsulated rPsaA. These results suggest that rPsaA coencapsulated with CTB can be used as an oral vaccine to induce cross-protective immunity for the prevention of pneumococcal infection. PMID- 11854196 TI - Genomic localization of a T serotype locus to a recombinatorial zone encoding extracellular matrix-binding proteins in Streptococcus pyogenes. AB - Streptococcus pyogenes is an important bacterial pathogen afflicting humans. A striking feature is its extraordinary biological diversity, evident in the wide range of diseases it can cause and the antigenic heterogeneity present on its surface. The T antigens form the basis of a major serological typing scheme that is often used as an alternative or supplement to M typing. Unlike M typing, the genetic basis for T typing is poorly understood. In this report, the tee6 gene is localized to a position approximately equal to 3.3 kb downstream from prtF1 (or sfbI), which encodes the Fn-binding protein, protein F, a key virulence factor. Comparison of this portion of the genome with those of four additional strains reveals the presence of genes encoding a collagen-binding protein (Cpa) and a second Fn-binding protein (PrtF2 or PfbpI). This chromosomal region--here designated the FCT region--is approximately 11 to 16 kb in length and is flanked at both ends by long stretches of highly conserved sequence. For each of the five strains, the FCT region contains a unique combination of semiconserved loci, indicative of extensive intergenomic recombination. The data provide evidence that the highly recombinatorial FCT region of the S. pyogenes genome is under strong selection for change in response to the host environment. PMID- 11854197 TI - Increased activation and oligoclonality of peripheral CD8(+) T cells in the chronic human helminth infection alveolar echinococcosis. AB - Alveolar echinococcosis (AE) in humans is a chronic disease characterized by slowly expanding liver lesions. Cellular immunity restricts the spreading of the extracellular pathogen, but functional contributions of CD4(+) and CD8(+) T cells are not defined. Here we studied ex vivo the phenotype and function of circulating T-cell subsets in AE patients by means of flow cytometry, T-cell receptor spectratyping, and lymphocyte proliferation. AE patients with parasitic lesions displayed a significant increase of activation of predominantly CD8(+) T cells compared to healthy controls and AE patients without lesions. In vitro, proliferative T-cell responses to polyclonal stimulation with recall antigens and Echinococcus multilocularis vesicular fluid antigen were sustained during chronic persisting infection in all AE patients. Only in AE patients with parasitic lesions did T-cell receptor spectratyping reveal increased oligoclonality of CD8(+) but not CD4(+) T cells, suggesting a persistent antigenic drive for CD8(+) T cells with subsequent proliferation of selected clonotypes. Thus, our data provide strong evidence for an active role of CD8(+) T cells in AE. PMID- 11854195 TI - Shigella flexneri Interactions with the Basolateral Membrane Domain of Polarized Model Intestinal Epithelium: Role of Lipopolysaccharide in Cell Invasion and in Activation of the Mitogen-Activated Protein Kinase ERK. AB - An early step governing Shigella flexneri pathogenesis is the invasion of the colonic epithelium from the basolateral surface followed by disruption of the colonic epithelial barrier. Despite recent insight into S. flexneri-host interactions, much remains to be determined regarding the nature of the initial contact between S. flexneri and the host epithelial basolateral membrane domain. Since the lipopolysaccharide (LPS) is located at the outermost part of the bacterial membrane, we considered that this component might be used by S. flexneri to attach to the basolateral surface of the intestinal epithelium and promote a proinflammatory response. Therefore, polarized human T84 intestinal epithelial cells were infected from the basolateral surface with either wild-type S. flexneri or one of its isogenic LPS-defective strains with mutations in either rfc, rfaL, or galU. We found that both adherence to and internalization into the basolateral surface of a polarized intestinal epithelium with S. flexneri were highly dependent on the length of the LPS (i.e., rfc > rfaL > galU). Furthermore, the addition of the anti-inflammatory LPS (RsDPLA) considerably decreased the invasion profile of wild-type S. flexneri by nearly 50%. Since LPS is associated with host inflammation, we further examined whether this molecule was involved in Shigella-induced inflammatory events. We found that S. flexneri LPS plays an important role in mediating epithelial-derived signaling, which leads to directed migration of polymorphonuclear leukocytes across model intestinal epithelium. This signaling most likely involves the activation of the mitogen-activated protein kinase extracellular regulated kinase. Thus, our findings have important implications on the understanding of the mechanisms by which S. flexneri can elicit mucosal inflammation. PMID- 11854198 TI - Transcript heterogeneity of the p44 multigene family in a human granulocytic ehrlichiosis agent transmitted by ticks. AB - Human granulocytic ehrlichiosis (HGE) is an emerging tick-borne zoonosis caused by a strain of Anaplasma phagocytophila called the HGE agent, an obligatory intracellular bacterium. The agent expresses immunodominant 44-kDa outer membrane proteins (P44s) encoded by a multigene family. The present study established an experimental process for transmission of the HGE agent from infected mice (a reservoir model) to nymphal Ixodes scapularis ticks (a biological vector) and subsequently to horses (a patient model) by the adult infected ticks. Overall, a total of 20 different p44 transcripts were detected in the mammals, ticks, and cell cultures. Among them, a transcript from a p44-18 gene was major at acute stage in mice and horses but minor in ticks. Both mRNA and protein produced from the p44-18 gene were detected in the HGE agent cultivated in HL-60 cells at 37 degrees C, but their expression levels decreased in the organisms cultivated at 24 degrees C, suggesting that temperature is one of the factors that influence the expression of members of the p44 multigene family. Several additional p44 transcripts that were not detected in the mammals at the acute stage of infection were detected in ticks. Phylogenetic analysis of the 20 different p44 transcripts revealed that the major transcripts found in mammals and ticks were distinct, suggesting a difference in surface properties between populations of the HGE agent in different host environments. The present study provides new information for understanding the role of the p44 multigene family in transmission of the HGE agent between mammals and ticks. PMID- 11854199 TI - Modulation of gamma interferon-induced major histocompatibility complex class II gene expression by Porphyromonas gingivalis membrane vesicles. AB - Gamma interferon (IFN-gamma)-induced endothelial cells actively participate in initiating immune responses by interacting with CD4(+) T cells via class II major histocompatibility complex (MHC) surface glycoproteins. Previously, Porphyromonas gingivalis membrane vesicles were shown to selectively inhibit IFN-gamma-induced surface expression of HLA-DR molecules by human umbilical cord vascular endothelial cells. In this study, we demonstrated an absence of HLA-DR alpha mRNA from IFN-gamma-induced cells in the presence of P. gingivalis membrane vesicles by using reverse transcriptase-PCR and Southern blotting. Vesicles also prevented transcription of the gene encoding class II transactivator, a transactivator protein required for IFN-gamma-induced expression of MHC class II genes. In addition, the effects of vesicles on IFN-gamma signal transduction involving Jak and Stat proteins were characterized by using immunoprecipitation and Western blot analyses. Jak1 and Jak2 proteins could not be detected in endothelial cells treated with membrane vesicles. Consequently, IFN-gamma-induced phosphorylation of Jak1, Jak2, and Stat1 alpha proteins was prevented. The class II-inhibitory effect of the membrane vesicles could be eliminated by heating vesicles at 100 degrees C for 30 min or by treating them with a cysteine proteinase inhibitor. This indicates that the cysteine proteinases were most likely responsible for the absence of Jak proteins observed in vesicle-treated cells. The observed increased binding of radiolabeled IFN-gamma to vesicle-treated cells suggests that vesicles may also modulate the IFN-gamma interactions with the cell surface. However, no evidence was obtained demonstrating that vesicles affected the expression of IFN gamma receptors. Thus, P. gingivalis membrane vesicles apparently inhibited IFN gamma-induced MHC class II by disrupting the IFN-gamma signaling transduction pathway. Vesicle-inhibited class II expression also occurred in other IFN-gamma inducible cells. This suggested that the ability of P. gingivalis membrane vesicles to modulate antigen presentation by key cells may be an important mechanism used by this particular bacterium to escape immunosurveillance, thereby favoring its colonization and invasion of host tissues. PMID- 11854200 TI - Membrane localization of the S1 subunit of pertussis toxin in Bordetella pertussis and implications for pertussis toxin secretion. AB - Pertussis toxin is secreted from Bordetella pertussis with the assistance of the Ptl transport system, a member of the type IV family of macromolecular transporters. The S1 subunit and the B oligomer combine to form the holotoxin prior to export from the bacterial cell, although the site of assembly is not known. To better understand the pathway of pertussis toxin assembly and secretion, we examined the subcellular location of the S1 subunit, expressed with or without the B oligomer and the Ptl proteins. In wild-type B. pertussis, the majority of the S1 subunit that remained cell associated localized to the bacterial membranes. In mutants of B. pertussis that do not express pertussis toxin and/or the Ptl proteins, full-length S1, expressed from a plasmid, partitioned almost entirely to the bacterial membranes. Several lines of evidence strongly suggest that the S1 subunit localizes to the outer membrane of B. pertussis. First, we found that membrane-bound full-length S1 was almost completely insoluble in Triton X-100. Second, recombinant S1 previously has been shown to localize to the outer membrane of Escherichia coli (J. T. Barbieri, M. Pizza, G. Cortina, and R. Rappuoli, Infect. Immun. 58:999-1003, 1990). Third, the S1 subunit possesses a distinctive amino acid motif at its carboxy terminus, including a terminal phenylalanine, which is highly conserved among bacterial outer membrane proteins. By using site-directed mutagenesis, we determined that the terminal phenylalanine is critical for stable expression of the S1 subunit. Our findings provide evidence that prior to assembly with the B oligomer and independent of the Ptl proteins, the S1 subunit localizes to the outer membrane of B. pertussis. Thus, outer membrane-bound S1 may serve as a nucleation site for assembly with the B oligomer and for interactions with the Ptl transport system. PMID- 11854201 TI - Structure of Campylobacter jejuni lipopolysaccharides determines antiganglioside specificity and clinical features of Guillain-Barre and Miller Fisher patients. AB - Ganglioside mimicry in the lipopolysaccharide (LPS) fraction of Campylobacter jejuni isolated from Guillain-Barre syndrome (GBS) and Miller Fisher syndrome (MFS) patients was compared with isolates from patients with an uncomplicated enteritis. The antibody response to C. jejuni LPS and gangliosides in neuropathy patients and controls was compared as well. LPS from GBS and MFS-associated isolates more frequently contained ganglioside-like epitopes compared to control isolates. Almost all neuropathy patients showed a strong antibody response against LPS and multiple gangliosides in contrast to enteritis patients. Isolates from GBS patients more frequently had a GM1-like epitope than isolates from MFS patients. GQ1b-like epitopes were present in all MFS-associated isolates and was associated with anti-GQ1b antibody reactivity and the presence of oculomotor symptoms. These results demonstrate that the expression of ganglioside mimics is a risk factor for the development of post-Campylobacter neuropathy. This study provides additional evidence for the hypothesis that the LPS fraction determines the antiganglioside specificity and clinical features in post-Campylobacter neuropathy patients. PMID- 11854202 TI - Identification and characterization of hsa, the gene encoding the sialic acid binding adhesin of Streptococcus gordonii DL1. AB - Oral colonization by Streptococcus gordonii, an important cause of subacute bacterial endocarditis, involves bacterial recognition of sialic acid-containing host receptors. The sialic acid-binding activity of this microorganism was previously detected by bacterium-mediated hemagglutination and associated with a streptococcal surface component identified as the Hs antigen. The gene for this antigen (hsa) has now been cloned in Escherichia coli, and its expression has been detected by colony immunoblotting with anti-Hs serum. Mutants of S. gordonii containing hsa inactivated by the insertion of an erythromycin resistance gene or deletion from the chromosome were negative for Hs-immunoreactivity, bacterium mediated hemagglutinating activity, and adhesion to alpha 2-3-linked sialoglycoconjugates. The deletion in the latter mutants was complemented by plasmid-borne hsa, resulting in Hs antigen production and the restoration of cell surface sialic acid-binding activity. The hsa gene encodes a 203-kDa protein with two serine-rich repetitive regions in its 2,178-amino-acid sequence. The first serine-rich region occurs within the amino-terminal region of the molecule, between different nonrepetitive sequences that may be associated with sialic acid binding. The second serine-rich region, which is much longer than the first, is highly repetitive, containing 113 dodecapeptide repeats with a consensus sequence of SASTSASVSASE. This long repetitive region is followed by a typical gram positive cell wall anchoring region at the carboxyl-terminal end. Thus, the predicted properties of Hsa, which suggest an amino-terminal receptor-binding domain attached to the cell surface by a molecular stalk, are consistent with the identification of this protein as the sialic acid-binding adhesin of S. gordonii DL1. PMID- 11854203 TI - Use of bacteriophage Ba1 to identify properties associated with Bordetella avium virulence. AB - Bordetella avium causes bordetellosis, an upper respiratory disease of birds. Commercially raised turkeys are particularly susceptible. We report here on the use of a recently described B. avium bacteriophage, Ba1, as a tool for investigating the effects of lysogeny and phage resistance on virulence. We found that lysogeny had no effect on any of the in vivo or in vitro measurements of virulence we employed. However, two-thirds (six of nine) spontaneous phage resistant mutants of our virulent laboratory strain, 197N, were attenuated. Phage resistance was associated, in all cases, with an inability of the mutants to bind phage. Further tests of the mutants revealed that all had increased sensitivities to surfactants, and increased amounts of incomplete (O-antigen-deficient) lipopolysaccharide (LPS) compared to 197N. Hot phenol-water-extracted 197N LPS inactivated phage in a specific and dose-dependent manner. Acid hydrolysis and removal of lipid A had little effect upon the ability of isolated LPS to inactivate Ba1, suggesting that the core region and possibly the O antigen were required for phage binding. All of the mutants, with one exception, were significantly more sensitive to naive turkey serum and, without exception, significantly less able to bind to tracheal rings in vitro than 197N. Interestingly, the three phage-resistant mutants that remained virulent appeared to be O antigen deficient and were among the mutants that were the most serum sensitive and least able to bind turkey tracheal rings in vitro. This observation allowed us to conclude that even severe defects in tracheal ring binding and serum resistance manifested in vitro were not necessarily indicative of attenuation and that complete LPS may not be required for virulence. PMID- 11854204 TI - Clonal diversity of Escherichia coli colonizing stools and urinary tracts of young girls. AB - Intestinal carriage of Escherichia coli in prepubertal girls without a history of urinary tract infection was examined by collecting weekly stools and periurethral and urine samples over 3 to 4 weeks of study. Dominant and minor clones were defined by grouping 28 E. coli isolates into clonal types. Multiple enteric clones of E. coli, which changed week to week, were found in the 13 girls during the study (median, 3 clones/girl; range, 1 to 16 clones/girl). Dominance of an enteric clone did not predict persistence in the stool. In only 10 (34%) of the 29 episodes in which a dominant clone present in one weekly sample could have been detected the following week did it persist as the dominant clone in the next weekly sample. In 5 (17%) of the 29 episodes, a dominant clone found in one weekly sample was classified as a minor clone the next week. Both dominant and minor clones were observed to colonize the urinary tract. However, when colonization of the periurethra or bladder urine occurred, it was brief and often did not reflect the dominant stool flora from the same week. In fact, in only 40% of episodes was a clone that was detected either on the periurethra or in the urine also recovered from the stool the same week. Our findings suggest that the intestinal flora of healthy girls is multiclonal with frequent fluctuations in composition. PMID- 11854205 TI - Genetic variability and stability of Anaplasma phagocytophila msp2 (p44). AB - Anaplasma (Ehrlichia) phagocytophila's major immunodominant surface protein antigen, Msp2 (P44, 44-kDa antigen), is encoded by a family of paralogous genes characterized by conserved sequences flanking a hypervariable region. The antigenic profiles of most strains of A. phagocytophila are different, and the differences are principally related to Msp2 expression. To date, multiple unique msp2 gene paralogs have been found in A. phagocytophila isolates, but the overall number in the genome of a single strain is not yet known. Changes in msp2 expression may be related to antigenic variability; thus, we examined the minimal complement of msp2 genes or pseudogenes in two strains of A. phagocytophila and the number of transcriptionally active msp2 gene paralogs during low-passage, steady-state, in vitro propagation. Of 15 BDS strain clones, 1 had a hypervariable region identical to the region in a clone obtained from a BDS strain genomic library previously prepared from organisms after only two horse passages. When 124 Webster strain clones were examined, 18 unique hypervariable regions were identified. Of 64 Webster strain cDNA clones, 56 (87.5%) were derived from a single gene, and transcripts from six additional msp2 genes were also identified. The sequences of several hypervariable regions that were > or = 97% similar to regions present in other strains were identified by performing a BLAST analysis of sequences deposited in the GenBank database. These findings suggest that antigenic variability results from transcription of one or a few of the multiple paralogs and not from genetic instability that results in random accumulated mutations, although the possibility that gene recombination plays a role cannot be eliminated. The predominant Msp2 pattern in vitro is determined by transcription from a single gene. PMID- 11854206 TI - Recombinant Dirofilaria immitis polyprotein that stimulates murine B cells to produce nonspecific polyclonal immunoglobulin E antibody. AB - Nonspecific immunoglobulin E (IgE) production is an event characteristically observed in parasitic helminth infections, but its mechanisms are still unclear. To define these mechanisms, we prepared a recombinant Dirofilaria immitis protein (rDiAg) and assessed its effect on nonspecific IgE production. rDiAg preferentially induced nonspecific IgE production, without eliciting specific IgE production, as well as a Th2-type cytokine profile (high interleukin-4 [IL-4] and IL-10 production but low gamma interferon production) in BALB/c mice. rDiAg significantly elicited the proliferative response of naive B cells. This response was not abolished by polymyxin B, an inhibitor of lipopolysaccharide (LPS), and rDiAg normally expanded splenic B cells from LPS nonresponder C3H/HeJ mice. Thus, the mitogenic effect of rDiAg was not due to LPS contamination. rDiAg also enhanced levels of CD23 expression on splenic B cells. Splenic B cells produced marked levels of IgE when cultured with the combination of rDiAg and IL-4 (rDiAg IL-4), whereas peritoneal B cells produced negligible levels of IgE. rDiAg-IL-4 induced IgE production by splenic B cells was synergistically increased by coculture with peritoneal B cells. rDiAg-driven IL-10 secretion was higher in peritoneal B cells than in splenic B cells. IgE production by splenic B cells cocultured with peritoneal B cells was decreased to a level comparable to that by splenic B cells in the presence of a neutralizing anti-IL-10 monoclonal antibody. Collectively, these results suggest that rDiAg-induced polyclonal expansion and IgE class switching of splenic B cells contribute to nonspecific IgE production and that these responses are enhanced by peritoneal B-cell-derived IL-10. PMID- 11854207 TI - Effect of Mycobacterium bovis BCG vaccination on interleukin-1 beta and RANTES mRNA expression in guinea pig cells exposed to attenuated and virulent mycobacteria. AB - The effect of Mycobacterium bovis BCG vaccination on interleukin-1 beta (IL-1 beta) or regulated-upon-activation, normally T-cell-expressed and -secreted chemokine (RANTES) mRNA expression in guinea pig spleen cells stimulated with concanavalin A, lipopolysaccharide (LPS), phorbol myristate acetate (PMA) plus ionomycin, or purified protein derivative (PPD) was studied in vitro. Similarly, peritoneal exudate cell-derived macrophages from naive and BCG-vaccinated guinea pigs were infected with M. bovis BCG, Mycobacterium avium, the attenuated Mycobacterium tuberculosis H37Ra strain, or virulent strains H37Rv and Erdman of M. tuberculosis. Total RNA was subjected to Northern blot analysis using probes generated from guinea pig IL-1 beta or RANTES cDNA. Although IL-1 beta and RANTES mRNA could be detected in the spleen cells from naive animals stimulated with LPS or PMA plus ionomycin, the levels were significantly enhanced after BCG vaccination. mRNA expression was also elevated in macrophages infected with live mycobacteria after BCG vaccination. However, macrophages infected with the virulent H37Rv strain of M. tuberculosis showed 75 to 90% reductions in IL-1 beta expression and 25 to 60% reductions in RANTES mRNA expression compared with macrophages infected with the attenuated H37Ra strain. The IL-1 beta mRNA levels peaked as soon as 1 h after PPD stimulation and 4 h after M. tuberculosis H37Rv infection of macrophages. In contrast, RANTES mRNA expression was delayed until 48 h after infection. These results indicate that molecular mediators produced in response to various stimuli associated with protective immunity against mycobacteria are upregulated after BCG vaccination; however, a significantly weaker response was observed with virulent M. tuberculosis. These initial studies indicate that BCG vaccination has a positive effect on IL-1 beta and RANTES mRNA expression by host cells in a highly relevant animal tuberculosis model. PMID- 11854208 TI - Identification of major outer surface proteins of Streptococcus agalactiae. AB - To identify the major outer surface proteins of Streptococcus agalactiae (group B streptococcus), a proteomic analysis was undertaken. An extract of the outer surface proteins was separated by two-dimensional electrophoresis. The visualized spots were identified through a combination of peptide sequencing and reverse genetic methodologies. Of the 30 major spots identified as S. agalactiae specific, 27 have been identified. Six of these proteins, previously unidentified in S. agalactiae, were sequenced and cloned. These were ornithine carbamoyltransferase, phosphoglycerate kinase, nonphosphorylating glyceraldehyde 3-phosphate dehydrogenase, purine nucleoside phosphorylase, enolase, and glucose 6-phosphate isomerase. Using a gram-positive expression system, we have overexpressed two of these proteins in an in vitro system. These recombinant, purified proteins were used to raise antisera. The identification of these proteins as residing on the outer surface was confirmed by the ability of the antisera to react against whole, live bacteria. Further, in a neonatal-animal model system, we demonstrate that some of these sera are protective against lethal doses of bacteria. These studies demonstrate the successful application of proteomics as a technique for identifying vaccine candidates. PMID- 11854209 TI - Mutational analysis of ganglioside GM(1)-binding ability, pentamer formation, and epitopes of cholera toxin B (CTB) subunits and CTB/heat-labile enterotoxin B subunit chimeras. AB - Variants of cholera toxin B subunit (CTB) were made by bisulfite- and oligonucleotide-directed mutagenesis of the ctxB gene. Variants were screened by a radial passive immune hemolysis assay (RPIHA) for loss of binding to sheep erythrocytes (SRBC). Variant CTBs were characterized for the formation of immunoreactive pentamers, the ability to bind ganglioside GM(1) in vitro, and reactivity with a panel of monoclonal anti-CTB antibodies. Substitutions at eight positions (i.e., positions 22, 29, 36, 45, 64, 86, 93, and 100) greatly reduced the yield of immunoreactive CTB. RPIHA-negative substitution variants that formed immunoreactive pentamers were obtained for residues 12, 33, 36, 51, 52 + 54, 91, and 95. Tyrosine-12 was identified as a novel residue important for GM(1) binding since, among all of the novel variants isolated with altered RPIHA phenotypes, only CTB with aspartate substituted for tyrosine at position 12 failed to bind significantly to ganglioside GM(1) in vitro. In contrast, CTB variants with single substitutions for several other residues (Glu-51, Lys-91, and Ala-95) that participate in GM(1) binding, based on the crystal structure of CTB and the oligosaccharide of GM(1), were not appreciably altered in their ability to bind GM(1) in vitro, even though they showed altered RPIHA phenotypes and did not bind to SRBC. Hybrid B genes made by fusing ctxB and the related Escherichia coli heat labile enterotoxin eltB genes at codon 56 produced CTB variants that had 7 or 12 heat-labile enterotoxin B residue substitutions in the amino or carboxyl halves of the monomer, respectively, each of which which also bound GM(1) as well as wild-type CTB. This collection of variant CTBs in which 47 of the 103 residues were substituted was used to map the epitopes of nine anti-CTB monoclonal antibodies (MAbs). Each MAb had a unique pattern of reactivity with the panel of CTB variants. Although no two of the epitopes recognized by different MAbs were identical, most of the single amino acid substitutions that altered the immunoreactivity of CTB affected more that one epitope. The tertiary structures of the epitopes of these anti-CTB MAbs are highly conformational and may involve structural elements both within and between CTB monomers. Substitution of valine for alanine at positions 10 and 46 had dramatic effects on the immunoreactivity of CTB, affecting epitopes recognized by eight or six MAbs, respectively. PMID- 11854211 TI - Cloned Shiga toxin 2 B subunit induces apoptosis in Ramos Burkitt's lymphoma B cells. AB - The Shiga toxins (Stx1 and Stx2), produced by Shigella dysenteriae type 1 and enterohemorrhagic Escherichia coli, consist of one A subunit and five B subunits. The Stx1 and Stx2 B subunits form a pentameric structure that binds to globotriaosylceramide (Gb3-Cer) receptors on eukaryotic cells and promotes endocytosis. The A subunit then inhibits protein biosynthesis, which triggers apoptosis in the affected cell. In addition to its Gb3-Cer binding activity, the data in the following report demonstrate that the Stx2 B pentamer induces apoptosis in Ramos Burkitt's lymphoma B cells independently of A subunit activity. Apoptosis was not observed in A subunit-free preparations of the Stx1 B pentamer which competitively inhibited Stx2 B pentamer-mediated apoptosis. The pancaspase inhibitor, Z-VAD-fmk, prevented apoptosis in Ramos cells exposed to the Stx2 B subunit, Stx1 or Stx2. Brefeldin A, an inhibitor of the Golgi transport system, also prevented Stx2 B subunit-mediated apoptosis. These observations suggest that the Stx2 B subunit must be internalized, via Gb3-Cer receptors, to induce Ramos cell apoptosis. Moreover, unlike the two holotoxins, Stx2 B subunit-mediated apoptosis does not involve inhibition of protein biosynthesis. This study provides further insight into the pathogenic potential of this family of potent bacterial exotoxins. PMID- 11854210 TI - Human gingival CD14(+) fibroblasts primed with gamma interferon increase production of interleukin-8 in response to lipopolysaccharide through up regulation of membrane CD14 and MyD88 mRNA expression. AB - Gamma interferon (IFN-gamma)-primed human gingival fibroblasts (HGF) have been shown to produce higher levels of interleukin-8 (IL-8) upon stimulation with bacterial products and inflammatory cytokines than nonprimed controls. In this study, we examined whether priming of HGF with IFN-gamma up-regulates IL-8 production by the cells in response to purified lipopolysaccharide (LPS). The priming effect of IFN-gamma was clearly observed in the high-CD14-expressing (CD14(high)) HGF but not in the low-CD14-expressing (CD14(low)) HGF. The CD14(high) HGF were most effectively primed with IFN-gamma (1,000 IU/ml) for 72 h. To elucidate the mechanism of the priming effects of IFN-gamma for the LPS response by HGF, we examined whether IFN-gamma regulated expression of CD14, Toll like receptor 2 (TLR2), TLR4, MD-2, and MyD88, all of which are molecules suggested to be associated with LPS signaling. In CD14(high) HGF, IFN-gamma markedly up-regulated CD14 and MyD88 but not TLR4 protein and MD-2 mRNA expression, while in CD14(low) HGF, IFN-gamma slightly increased MyD88 and scarcely affected CD14, TLR4 protein, and MD-2 mRNA levels. LPS-induced IL-8 production by IFN-gamma-primed CD14(high) HGF was significantly inhibited by monoclonal antibodies (MAbs) against CD14 and TLR4, but not by an anti-TLR2 MAb. These findings suggested that IFN-gamma primed CD14(high) HGF to enhance production of IL-8 in response to LPS through augmentation of the CD14-TLR system, where the presence of membrane CD14 was indispensable for the response of HGF to LPS. PMID- 11854212 TI - Fibronectin facilitates Mycobacterium tuberculosis attachment to murine alveolar macrophages. AB - Mycobacterium tuberculosis remains a major cause of pulmonary infection worldwide. Attachment of M. tuberculosis organisms to alveolar macrophages (AMs) represents the earliest phase of primary infection in pulmonary tuberculosis. In this study fibronectin (Fn), an adhesive protein, is shown to bind M. tuberculosis organisms and facilitates attachment of M. tuberculosis to murine AMs. A monoclonal antibody (MAb) specific to the heparin binding domain (HBD) of Fn decreases (125)I-Fn binding to M. tuberculosis; whereas MAbs specific to either the cell binding domain (CBD) or the gelatin binding domain (GBD) have no effect on Fn binding to M. tuberculosis. In the presence of exogenous Fn (10 microg/ml) M. tuberculosis attachment to AMs increased significantly from control levels (means +/- standard errors of the means) of 11.5% +/- 1.1% to 44.2% +/- 4.2% (P < 0.05). Fn-enhanced attachment was significantly decreased from 44.2% +/ 4.2% to 10.8% +/- 1.2% (P < 0.05) in the presence of anti-Fn polyclonal antibodies. The attachment is also inhibited in the presence of MAbs specific for the HBD and CBD, whereas MAbs specific to GBD did not affect the attachment. Further, an Fn cell binding peptide, Arg-Gly-Asp-Ser (RGDS), decreased the attachment from 44.2% +/- 4.2% to 15.3% +/- 1.2% (P < 0.05), whereas addition of a control peptide, Arg-Gly-Glu-Ser (RGES) did not affect the attachment (40.5% +/ 1.8%). These results suggest that Fn-mediated attachment of M. tuberculosis can occur through the binding of Fn to the AM via the CBD and to M. tuberculosis organisms via the HBD. PMID- 11854213 TI - Cross-reactive polyclonal antibodies to the inner core of lipopolysaccharide from Neisseria meningitidis. AB - Sera from mice immunized with native or detergent-extracted outer membrane vesicles derived from lipopolysaccharide (LPS) mutant 44/76(Mu-4) of Neisseria meningitidis were analyzed for antibodies to LPS. The carbohydrate portion of 44/76(Mu-4) LPS consists of the complete inner core, Glc beta 1-->4[GlcNAc alpha 1-->2Hep alpha 1-->3]Hep alpha 1-->5KDO[4-->2 alpha KDO]. Immunoblot analysis revealed that some sera contained antibodies to wild-type LPS which has a fully extended carbohydrate chain of immunotype L3,7, as well as to the homologous LPS. Sera reacted only weakly to LPS from 44/76(Mu-3), which lacks the terminal glucose of the inner core. No binding to more truncated LPS was observed. Consequently, the cross-reactive epitopes are expressed mainly by the complete inner core. Dephosphorylation of wild-type LPS abolished antibody binding to LPS in all but one serum. Thus, at least two specificities of cross-reactive antibodies exist: one is dependent on phosphoethanolamine groups in LPS, and one is not. Detection of these cross-reactive antibodies strongly supports the notion that epitopes expressed by meningococcal LPS inner core are also accessible to antibodies when the carbohydrate chain is fully extended. Also, these inner core epitopes are sufficiently immunogenic to induce antibody levels detectable in polyclonal antibody responses. Meningococci can escape being killed by antibodies to LPS that bind only to a specific LPS variant, by altering the carbohydrate chain length. Cross-reactive antibodies may prevent such escape. Therefore, inner core LPS structures may be important antigens in future vaccines against meningococcal disease. PMID- 11854214 TI - Characterization of humoral and cellular immune responses elicited by meningococcal carriage. AB - In order to study the immune response elicited by asymptomatic carriage of Neisseria meningitidis, samples of serum, peripheral blood mononuclear cells (PBMCs), and saliva were collected from a cohort of more than 200 undergraduate students in Nottingham, United Kingdom, who were subject to high rates of acquisition and carriage of meningococci. Serum immunoglobulin G levels were elevated following increases in the rate of carriage, and these responses were specific for the colonizing strains. In order to investigate T-cell responses, PBMCs from 15 individuals were stimulated with a whole-cell lysate of the H44/76 meningococcal strain (B:15:P1.7,16), stained to detect cell surface markers and intracellular cytokines, and examined by flow cytometry. The cells were analyzed for expression of CD69 (to indicate activation), gamma interferon (IFN-gamma) (a representative T-helper 1 subset [Th1]-associated cytokine), and interleukin-5 (IL-5) (a Th2-associated cytokine). Following a brief meningococcal stimulation, the numbers of CD69(+) IFN-gamma(+) CD56/16(+) NK cells were much higher than cytokine-positive CD4(+) events. Both IFN-gamma(+) and IL-5(+) events were detected among the CD69(+) CD4(+) population, leading to the conclusion that an unbiased T-helper subset response was elicited by meningococcal carriage. PMID- 11854215 TI - Retrieving biological activity from LukF-PV mutants combined with different S components implies compatibility between the stem domains of these staphylococcal bicomponent leucotoxins. AB - Bicomponent leucotoxins, such as Panton-Valentine leucocidin, are composed of two classes of proteins, a class S protein such as LukS-PV, which bears the cell membrane binding function, and a class F protein such as LukF-PV, which interacts to form a bipartite hexameric pore. These leucotoxins induce cell activation, linked to a Ca(2+) influx, and pore formation as two consecutive and independently inhibitable events. Knowledge of the LukF-PV monomer structure has indicated that the stem domain is folded into three antiparallel beta-strands in the water-soluble form and has to refold into a transmembrane beta-hairpin during pore formation. To investigate the requirements for the cooperative assembly of the stems of the S and F components to produce biological activity, we introduced multiple deletions or single point mutations into the stem domains of LukF-PV and HlgB. While the binding of the mutated proteins was weakly dependent on these changes, Ca(2+) influx and pore formation were affected differently, confirming that they are independent events. Ca(2+) entry into human polymorphonuclear cells requires oligomerization and may follow the formation of a prepore. The activity of some of the LukF-PV mutants, carrying the shorter deletions, was actually improved. This demonstrated that a crucial event in the action of these toxins is the transition of the prefolded stem into the extended beta-hairpins and that this step may be facilitated by small deletions that remove some of the interactions stabilizing the folded structure. PMID- 11854216 TI - Contribution of fibronectin-binding protein to pathogenesis of Streptococcus suis serotype 2. AB - In the present study we investigated the role of the fibronectin (FN)- and fibrinogen (FGN)-binding protein (FBPS) in the pathogenesis of Streptococcus suis serotype 2 in piglets. The complete gene encoding FBPS from S. suis serotype 2 was cloned in Escherichia coli and sequenced. The occurrence of the gene in various serotypes was analyzed by hybridization studies. The FBPS protein was expressed in E. coli and purified, and binding to human FN and FGN was demonstrated. The induction of antibodies in piglets was studied upon infection. An isogenic mutant unable to produce FBPS was constructed, and the levels of virulence of the wild-type and mutant strains were compared in a competitive infection model in young piglets. Organ cultures showed that FBPS was not required for colonization of the tonsils but that FBPS played a role in the colonization of the specific organs involved in an S. suis infection. Therefore, the FBPS mutant was considered as an attenuated mutant. PMID- 11854217 TI - Virulence gene identification by differential fluorescence induction analysis of Staphylococcus aureus gene expression during infection-simulating culture. AB - We have employed a strategy utilizing differential fluorescence induction (DFI) in an effort to identify Staphylococcus aureus genes whose products can be targeted for antimicrobial drug development. DFI allows identification of promoters preferentially active under given growth conditions on the basis of their ability to drive expression of a promoterless green fluorescent protein gene (gfp). A plasmid-based promoter trap library was constructed of 200- to 1,000-bp fragments of S. aureus genomic DNA fused to gfp, and clones with active promoters were isolated under seven different in vitro growth conditions simulating infection. Six thousand two hundred sixty-seven clones with active promoters were screened to identify those that exhibited differential promoter activity. Bioinformatic analysis allowed the identification of 42 unique operons, containing a total of 61 genes, immediately downstream of the differentially active putative promoters. Replacement mutations were generated for most of these operons, and the abilities of the resulting mutants to cause infection were assessed in two different murine infection models. Approximately 40% of the mutants were attenuated in at least one infection model. PMID- 11854218 TI - Dissociated linkage of cytokine-inducing activity and cytotoxicity to different domains of listeriolysin O from Listeria monocytogenes. AB - Listeriolysin O (LLO), a cholesterol-binding cytolysin of Listeria monocytogenes, exhibits cytokine-inducing and cytolytic activities. Because the cytolytic activity was abolished by cholesterol treatment but the cytokine-inducing activity was not, these activities appeared to be linked to different domains of the LLO molecule. In this study, we constructed recombinant full-length LLO (rLLO529) and various truncated derivatives and examined their cytolytic, cholesterol-binding, and gamma interferon (IFN-gamma)-inducing activities. rLLO529 exhibited both IFN-gamma-inducing and cytolytic activities. Four truncated rLLOs possessing different C termini, which did not exert either cytolytic or cholesterol-binding activity, stimulated IFN-gamma production in normal spleen cells. However, a truncated rLLO corresponding to domain 4 (rLLO416 529) did not exhibit IFN-gamma-inducing activity, whereas it did bind to immobilized cholesterol. In addition, though the hemolysis induced by rLLO529 was inhibited by rLLO416-529, such inhibition was not detected upon rLLO529-induced IFN-gamma production. These data indicated that domain 4 was responsible for binding of LLO to membrane cholesterol followed by oligomerization and pore formation by the entire LLO molecule. In contrast, the other part of LLO, corresponding to domain 1-3, was essential for IFN-gamma-inducing activity. These findings implied a novel aspect of the function of LLO as a bacterial modulin. PMID- 11854219 TI - Control of experimental Trypanosoma brucei infections occurs independently of lymphotoxin-alpha induction. AB - Trypanosome infections are marked by severe pathological features, including anemia, splenomegaly, and suppression of T-cell proliferation. We have used lymphotoxin-alpha-deficient (LT-alpha(-/-)) mice, as well as LT-alpha-tumor necrosis factor-double-deficient (LT-alpha(-/-) TNF(-/-)) mice, to analyze the contributions of these related cytokines in both induction of trypanosomosis associated immunopathology and infection control. Moreover, as the cytokine deficient mice used have no detectable lymph nodes and lack germinal-center formation upon immune stimulation, we have analyzed the functional importance of both the lymph nodes and spleen during experimental Trypanosoma brucei infections. First, we show that the absence of LT-alpha does not significantly alter early trypanosomosis development or pathology but does result in better control of late-stage parasitemia levels and slightly prolonged survival. This increased survival of infected LT-alpha(-/-) mice coincides with the appearance of increased chronic-stage anti-trypanosome immunoglobulin M (IgM)-IgG2a serum titers that are generated in the absence of functional peripheral lymphoid tissue and do not require germinal-center formation. Second, we show that splenectomized mice control their parasitemia to the same extent as fully immune-competent littermates. Finally, using LT-alpha(-/-) TNF(-/-) double-deficient mice, we show that in these mice T. brucei infections are very well controlled during the chronic infection stage and that infection-induced pathology is minimized. Together, these findings indicate that while increased IgM-IgG2a anti-trypanosome antibody titers (generated in the absence of LT-alpha, peripheral lymph nodes, and germinal-center formation) coincide with improved parasitemia control, it is TNF that has a major impact on trypanosomosis-associated immunopathology. PMID- 11854220 TI - Pulmonary inflammation induced by Pseudomonas aeruginosa lipopolysaccharide, phospholipase C, and exotoxin A: role of interferon regulatory factor 1. AB - Chronic pulmonary infection with Pseudomonas aeruginosa is common in cystic fibrosis (CF) patients. P. aeruginosa lipopolysaccharide (LPS), phosholipase C (PLC), and exotoxin A (ETA) were evaluated for their ability to induce pulmonary inflammation in mice following intranasal inoculation. Both LPS and PLC induced high levels of tumor necrosis factor alpha (TNF-alpha), interleukin 1 beta (IL-1 beta-6, gamma interferon (IFN-gamma), MIP-1 alpha MIP-2 in the lungs but did not affect IL-18 levels. ETA did not induce TNF-alpha and was a weak inducer of IL-1 beta, IL-6, macrophage inflammatory protein 1 alpha (MIP-1 alpha), and MIP-2. Remarkably, ETA reduced constitutive lung IL-18 levels. LPS was the only factor inducing IFN-gamma. LPS, PLC, and ETA all induced cell infiltration in the lungs. The role of interferon regulatory factor-1 (IRF-1) in pulmonary inflammation induced by LPS, PLC, and ETA was evaluated. When inoculated with LPS, IRF-1 gene knockout (IRF-1 KO) mice produced lower levels of TNF-alpha, IL-1 beta, and IFN gamma than did wild-type (WT) mice. Similarly, a milder effect of ETA on IL-1 beta and IL-18 was observed for IRF-1 KO than for WT mice. In contrast, the cytokine response to PLC did not differ between WT and IRF-1 KO mice. Accordingly, LPS and ETA, but not PLC, induced expression of IRF-1 mRNA. IRF-1 deficiency had no effect on MIP-1 alpha and MIP-2 levels and on cell infiltration induced by LPS, PLC, or ETA. Flow cytometric evaluation of lung mononuclear cells revealed strongly reduced percentages of CD8(+) and NK cells in IRF-1 KO mice compared to percentages observed for WT mice. These data indicate that different virulence factors from P. aeruginosa induce pulmonary inflammation in vivo and that IRF-1 is involved in some of the cytokine responses to LPS and ETA. PMID- 11854222 TI - A superoxide dismutase C mutant of Haemophilus ducreyi is virulent in human volunteers. AB - Haemophilus ducreyi produces a periplasmic copper-zinc superoxide dismutase (Cu Zn SOD), which is thought to protect the organism from exogenous reactive oxygen species generated by neutrophils during an inflammatory response. We had previously identified the gene, sodC, responsible for the production and secretion of Cu-Zn SOD and constructed an isogenic H. ducreyi strain with a mutation in the sodC gene (35000HP-sodC-cat). Compared to the parent, the mutant does not survive in the presence of exogenous superoxide (L. R. San Mateo, M. Hobbs, and T. H. Kawula, Mol. Microbiol. 27:391-404, 1998) and is impaired in the swine model of H. ducreyi infection (L. R. San Mateo, K. L. Toffer, P. E. Orndorff, and T. H. Kawula, Infect. Immun. 67:5345-5351, 1999). To test whether Cu-Zn SOD is important for bacterial survival in vivo, six human volunteers were experimentally infected with 35000HP and 35000HP-sodC-cat and observed for papule and pustule formation. Papules developed at similar rates at sites inoculated with the mutant or parent. The pustule formation rates were 75% (95% confidence intervals [CI], 43 to 95%) at 12 parent-inoculated sites and 67% (95% CI, 41 to 88%) at 18 mutant-inoculated sites (P = 0.47). There was no significant difference in levels of H. ducreyi recovery from mutant- and parent-inoculated biopsy sites. These results suggest that expression of Cu-Zn SOD does not play a major role in the survival of this pathogen in the initial stages of experimental infection of humans. PMID- 11854221 TI - Effects of Anaplasma phagocytophila on NADPH oxidase components in human neutrophils and HL-60 cells. AB - The human granulocytic ehrlichiosis agent, Anaplasma phagocytophila, resides and multiplies exclusively in cytoplasmic vacuoles of granulocytes. A. phagocytophila rapidly inhibits the superoxide anion (O(2)(-)) generation by human neutrophils in response to various stimuli. To determine the inhibitory mechanism, the influence of A. phagocytophila on protein levels and localization of components of the NADPH oxidase were examined. A. phagocytophila decreased levels of p22(phox), but not gp91(phox), p47(phox), p67(phox), or P40(phox) reactive with each component-specific antibody in human peripheral blood neutrophils and HL-60 cells. Double immunofluorescence labeling revealed that p47(phox), p67(phox), Rac2, and p22(phox) did not colocalize with A. phagocytophila inclusions in neutrophils or HL-60 cells, and p22(phox) levels were also reduced. A. phagocytophila did not prevent either membrane translocation of cytoplasmic p47(phox) and p67(phox) or phosphorylation of p47(phox) upon stimulation by phorbol myristate acetate. The inhibitory signals for O(2)(-) generation was independent of several signals required for A. phagocytophila internalization. These results suggest that rapid alteration in p22(phox) induced by binding of A. phagocytophila to neutrophils is involved in the inhibition of O(2)(-) generation. Absence of colocalization of NADPH oxidase components with the inclusion further protects A. phagocytophila from oxidative damage. PMID- 11854223 TI - Role of osteopontin in murine Lyme arthritis and host defense against Borrelia burgdorferi. AB - Several genetic loci in the mouse have been identified that regulate the severity of Lyme arthritis. The region of chromosome 5 including the osteopontin (OPN) gene (Opn) has been identified in intercross populations of C3H/HeN x C57BL/6 and C3H/HeJ x BALB/cAnN mice. OPN is of particular interest as it is involved in the maintenance and remodeling of tissue during inflammation, it regulates production of interleukin-10 (IL-10) and IL-12 (cytokines implicated in Lyme arthritis), it is necessary for host control of certain bacterial infections, and mice displaying different severities of Lyme arthritis possess different alleles of the OPN gene. Macrophages and splenocytes from OPN-deficient mice on mixed C57BL/6J-129S or inbred 129S backgrounds were stimulated with the Pam(3)Cys modified lipoprotein from Borrelia burgdorferi, OspA. OPN was not required for OspA-induced cytokine production; however, macrophages from 129S-Opn(-/-) mice displayed a reduced level of IL-10 production. OPN was also not required for resistance to severe arthritis, as B. burgdorferi-infected 129S-Opn(-/-) mice developed mild arthritis, as did their wild-type littermates. Arthritis was more severe in OPN-deficient mice on the mixed C57BL/6J-129S backgrounds than in inbred mice of either strain. This increase was most likely due to a gene(s) closely linked to Opn on chromosome 5 in conjunction with other randomly assorting genes. Deficiency in OPN did not influence the numbers of spirochetes in tissues from B. burgdorferi-infected mice, indicating OPN is not part of the host defense to this pathogen. Interestingly, there was no alteration in the B. burgdorferi-specific antibody isotypes in OPN-deficient mice, indicating that its effect on helper T-cell responses is not relevant to the host response to B. burgdorferi. PMID- 11854224 TI - The sortase SrtA of Listeria monocytogenes is involved in processing of internalin and in virulence. AB - Listeria monocytogenes is an intracellular gram-positive human pathogen that invades eucaryotic cells. Among the surface-exposed proteins playing a role in this invasive process, internalin belongs to the family of LPXTG proteins, which are known to be covalently linked to the bacterial cell wall in gram-positive bacteria. Recently, it has been shown in Staphylococcus aureus that the covalent anchoring of protein A, a typical LPXTG protein, is due to a cysteine protease, named sortase, required for bacterial virulence. Here, we identified in silico from the genome of L. monocytogenes a gene, designated srtA, encoding a sortase homologue. The role of this previously unknown sortase was studied by constructing a sortase knockout mutant. Internalin was used as a reporter protein to study the effects of the srtA mutation on cell wall anchoring of this LPXTG protein in L. monocytogenes. We show that the srtA mutant (i) is affected in the display of internalin at the bacterial surface, (ii) is significantly less invasive in vitro, and (iii) is attenuated in its virulence in the mouse. These results demonstrate that srtA of L. monocytogenes acts as a sortase and plays a role in the pathogenicity. PMID- 11854226 TI - Type III secretion of Salmonella enterica serovar Typhimurium translocated effectors and SseFG. AB - The type III secretion system (TTSS) encoded by Salmonella enterica serovar Typhimurium pathogenicity island 2 (SPI2) is employed by Salmonella enterica for interaction with host cells during the intracellular phase of pathogenesis. This TTSS secretes a set of SPI2-encoded proteins in vitro and translocates Salmonella serovar Typhimurium translocated effectors (STE) that are encoded by genes outside of SPI2 into host cells. Using an epitope-tagging approach, we analyzed secretion of proteins by the TTSS of SPI2 and identified SseF and SseG as further secreted substrate proteins. Three members of the STE family, SifA, SifB, and SseJ, were secreted under conditions that also induce secretion of SPI2-encoded substrate proteins. PMID- 11854225 TI - Osmolarity and pH growth conditions regulate fim gene transcription and type 1 pilus expression in uropathogenic Escherichia coli. AB - A comparative study was performed to determine the effects of pH, osmolarity, and human urine on the transcription of several fim genes, as well as the overall expression of type 1 pili. Several fim-lacZYA fusions were constructed on single copy plasmids to test a range of pHs and a range of osmolarities. Growth in acidic medium slightly reduced expression from all of the fim promoters (fimA, fimB, and fimE). Increased osmolarity in neutral-pH medium repressed fimA and fimB transcription by approximately 50% when 400 mM NaCl was used and nearly threefold when 800 mM NaCl was used, whereas fimE transcription rose slightly as the osmolarity increased. This effect was more pronounced in high-osmolarity acidic media; fimB and fimA expression decreased fivefold in growth media containing 800 mM NaCl compared to expression in growth media without added NaCl. Moreover, fimE expression doubled under the same high-osmolarity conditions compared to expression in a low-osmolarity acidic environment. When a fimB-lacZ or fimE-lacZ fusion was inserted into the chromosome of strain AAEC189, fimE expression changed slightly as the osmolarity increased, but fimB expression decreased by 50% in a low-pH high-osmolarity environment. When strain AAEC189 with either a plasmid-borne fimB-lacZ fusion or a plasmid-borne fimE-lacZ fusion was grown in human urine, similar changes in the levels of fimB and fimE expression were observed. Limiting-dilution reverse transcription-PCR confirmed that these changes in fim expression occurred in clinical isolates of uropathogenic Escherichia coli grown in media with different pHs and different osmolarities. Furthermore, the invertible switch region in uropathogenic strain NU149 shifted from favoring the phase-on position in a neutral-pH low-osmolarity environment to favoring the phase-off position in a low-pH high-osmolarity environment. Results obtained with an ompR mutant strain demonstrated that fimB expression was derepressed and that OmpR may neutralize repression by an acid response regulator of fimE expression in a low-pH environment. In addition, H-NS was verified to be important in regulation of fimB, but it had only a slight effect on fimE under the specific pH and osmotic growth conditions tested. Enzyme immunoassays with anti-type 1 pilus antibody and hemagglutination assays showed that fewer type 1 pili were detected with cells in a low-pH high-osmolarity environment. Together, these observations demonstrate that a combination of low pH and high osmolarity regulates the transcription of fim genes, which favors a shift in the invertible element to the phase-off orientation and a loss of type 1 pilus expression. Taken together, our data suggest that the environmental cues that we tested may regulate expression of type 1 pili in specific in vivo niches, such as murine kidneys and possibly human kidneys. PMID- 11854227 TI - Lymphocyte recruitment and protective efficacy against pulmonary mycobacterial infection are independent of the route of prior Mycobacterium bovis BCG immunization. AB - Mycobacterium tuberculosis infects humans through the lung, and immunity to this chronic infection is mediated primarily by CD4(+) T lymphocytes. Recently we have demonstrated that the recruitment of lymphocytes to the lung during primary aerosol M. tuberculosis infection in mice occurs predominantly through the interaction of alpha(4)beta(1) integrin on CD4(+) T cells and vascular cell adhesion molecule-1 on the pulmonary endothelium. To investigate the effect of route of immunization with Mycobacterium bovis BCG on the pattern of T-cell recruitment to the lung, we have analyzed the differences in expression of integrins on activated memory CD4(+) T cells infiltrating the lung following primary BCG immunization by aerosol, intravenous, and subcutaneous routes and after subsequent aerosol challenge with M. tuberculosis. There were marked differences in the patterns of recruitment of activated CD4(+) T cells to the lung following primary immunization by the three routes. Expansion of CD44(hi) CD62L(low) CD4(+) T cells in the lung occurred following aerosol and intravenous BCG immunizations, and the lymphocyte recruitment was proportional to the pulmonary bacterial load. The majority of infiltrating CD4(+) T cells expressed alpha(4)beta(1) integrin. On subsequent exposure to aerosol BCG rapid expansion of gamma interferon-secreting alpha(4)beta(1)(+) CD4(+) T cells occurred to the same extent in all immunized mice, regardless of the route of immunization. Similar expansion of alpha(4)beta(1)(+) CD4(+) memory T cells occurred following M. tuberculosis challenge. The three routes of BCG immunization resulted in the same level of protection against aerosol M. tuberculosis or BCG challenge in both the lungs and spleen. Therefore, recruitment of effector T lymphocytes and protective efficacy against pulmonary mycobacterial infection are independent of the route of prior BCG immunization. PMID- 11854228 TI - Induction of T helper type 1 and 2 responses to 19-kilodalton merozoite surface protein 1 in vaccinated healthy volunteers and adults naturally exposed to malaria. AB - Plasmodium falciparum malaria is a major cause of death in the tropics. The 19 kDa subunit of P. falciparum merozoite surface protein 1 (MSP-1(19)), a major blood stage vaccine candidate, is the target of cellular and humoral immune responses in animals and humans. In this phase I trial of MSP-1(19), immunization of nonexposed human volunteers with either of the two allelic forms of recombinant MSP-1(19) induced high levels of antigen-specific Th1 (gamma interferon) and Th2 (interleukin 4 [IL-4] and IL-10) type lymphokines. The adjustment of the antigen dose and number of immunizations regulated the level of specificity of immune responses and Th1/Th2 bias of responses induced by vaccination. Novel conserved and allelic T-cell epitopes which induced cross strain immune responses were identified. Importantly, responses to many of these novel epitopes were also present in adults exposed to malaria, both in east (Kenya) and west Africa (The Gambia). These data suggest that epitope-specific naturally acquired MSP-1(19) immune responses in endemic populations can be boosted by vaccination. PMID- 11854229 TI - Differential fluorescence induction analysis of Streptococcus pneumoniae identifies genes involved in pathogenesis. AB - Differential fluorescence induction (DFI) technology was used to identify promoters of Streptococcus pneumoniae induced under various in vitro and in vivo conditions. A promoter-trap library using green fluorescent protein as the reporter was constructed in S. pneumoniae, and the entire library was screened for clones exhibiting increased gfp expression under the chosen conditions. The in vitro conditions used were chosen to mimic aspects of the in vivo environment encountered by the pathogen once it enters a host: changes in temperature, osmolarity, oxygen, and iron concentration, as well as blood. In addition, the library was used to infect animals in three different models, and clones induced in these environments were identified. Several promoters were identified in multiple screens, and genes whose promoters were induced twofold or greater under the inducing condition were mutated to assess their roles in virulence. A total of 25 genes were mutated, and the effects of the mutations were assessed in at least two different infection models. Over 50% of these mutants were attenuated in at least one infection model. We show that DFI is a useful tool for identifying bacterial virulence factors as well as a means of elucidating the microenvironment encountered by pathogens upon infection. PMID- 11854230 TI - Extracellular antigens from Salmonella enteritidis induce effective immune response in mice after oral vaccination. AB - We have studied polyacryl starch microparticles as an adjuvant in oral vaccination in mice. Secreted antigens from Salmonella enterica serovar Enteritidis were administered covalently conjugated to microparticles, or as free antigens, orally or intramuscularly and evaluated for their immunogenicity and ability to elicit protective immune response against an oral challenge with live serovar Enteritidis. The highest immunoglobulin M (IgM)-plus-IgG titers were obtained in the groups immunized with antigen-conjugated microparticles. The subclass profile switched to a stronger Th1 influence in the oral groups after booster, while the intramuscular group showed a constant Th1/Th2 profile. A strong specific IgA response was seen in feces in the oral groups, which was further confirmed in an enzyme-linked immunospot assay. The delayed-type hypersensitivity test, as a measure of the cellular response, showed a significant increase in ear thickness in all the immunized groups, except for the group that received free antigen orally, compared to the nonimmunized group. The cytokines released from in vitro-stimulated spleens showed a strong gamma interferon response in all immunized groups. A significant reduction in CFU in liver and spleen was seen in the orally immunized groups compared to the nonimmunized group after oral challenge with serovar Enteritidis. Western blotting analysis with both sera and feces revealed that antibodies against three bands, 53, 56, and 60 kDa, dominated the oral groups, and an electrospray-mass spectroscopy analysis of these bands showed amino acid sequences coinciding with those of phase-1 flagellin and hook-associated protein 2. PMID- 11854232 TI - Role of fraction 1 antigen of Yersinia pestis in inhibition of phagocytosis. AB - Yersinia pestis, the causative agent of plague, expresses a capsule-like antigen, fraction 1 (F1), at 37 degrees C. F1 is encoded by the caf1 gene located on the large 100-kb pFra plasmid, which is unique to Y. pestis. F1 is a surface polymer composed of a protein subunit, Caf1, with a molecular mass of 15.5 kDa. The secretion and assembly of F1 require the caf1M and caf1A genes, which are homologous to the chaperone and usher protein families required for biogenesis of pili. F1 has been implicated to be involved in the ability of Y. pestis to prevent uptake by macrophages. In this study we addressed the role of F1 antigen in inhibition of phagocytosis by the macrophage-like cell line J774. The Y. pestis strain EV76 was found to be highly resistant to uptake by J774 cells. An in-frame deletion of the caf1M gene of the Y. pestis strain EV76 was constructed and found to be unable to express F1 polymer on the bacterial surface. This strain had a somewhat lowered ability to prevent uptake by J774 cells. Strain EV76C, which is cured for the virulence plasmid common to the pathogenic Yersinia species, was, as expected, much reduced in its ability to resist uptake. A strain lacking both the virulence plasmid and caf1M was even further hampered in the ability to prevent uptake and, in this case, essentially all bacteria (95%) were phagocytosed. Thus, F1 and the virulence plasmid-encoded type III system act in concert to make Y. pestis highly resistant to uptake by phagocytes. In contrast to the type III effector proteins YopE and YopH, F1 did not have any influence on the general phagocytic ability of J774 cells. Expression of F1 also reduced the number of bacteria that interacted with the macrophages. This suggests that F1 prevents uptake by interfering at the level of receptor interaction in the phagocytosis process. PMID- 11854231 TI - Intranasal immunization with pneumococcal conjugate vaccines with LT-K63, a nontoxic mutant of heat-Labile enterotoxin, as adjuvant rapidly induces protective immunity against lethal pneumococcal infections in neonatal mice. AB - Immunization with pneumococcal polysaccharides (PPS) conjugated to tetanus toxoid (TT) (Pnc-TT) elicits protective immunity in an adult murine pneumococcal infection model. To assess immunogenicity and protective immunity in early life, neonatal (1 week old) and infant (3 weeks old) mice were immunized intranasally (i.n.) or subcutaneously (s.c.) with Pnc-TT of serotype 1 (Pnc1-TT). Anti-PPS-1 and anti-TT immunoglobulin G (IgG) and IgM antibodies were measured in serum and saliva, and vaccine-induced protection was evaluated by i.n. challenge with serotype 1 pneumococci. Pnc1-TT was immunogenic in neonatal and infant mice when administered s.c. without adjuvant: a majority of the young mice were protected from bacteremia and a reduction of pneumococcal density in the lungs was observed, although antibody responses and protective efficacy remained lower than in adults. The addition of LT-K63, a nontoxic mutant of heat-labile enterotoxin, as adjuvant significantly enhanced PPS-1-specific IgG responses and protective efficacy following either s.c. or i.n. Pnc1-TT immunization. Mucosal immunization was particularly efficient in neonates, as a single i.n. dose of Pnc1-TT and LT K63 induced significantly higher PPS-1-specific IgG responses than s.c. immunization and was sufficient to protect neonatal mice against pneumococcal infections, whereas two s.c. doses were required to induce complete protection. In addition, i.n. immunization with Pnc1-TT and LT-K63 induced a vigorous salivary IgA response. This suggests that mucosal immunization with pneumococcal conjugate vaccines and LT-K63 may be able to circumvent some of the limitations of neonatal antibody responses, which are required for protective immunity in early life. PMID- 11854233 TI - Replication of Neisseria meningitidis within epithelial cells requires TonB dependent acquisition of host cell iron. AB - Neisseria meningitidis (meningococcus [MC]) is able to enter and replicate within epithelial cells. Iron, an essential nutrient for nearly all organisms, is an important determinant in the ability of MC to cause disease; however, its role in MC intracellular replication has not been investigated. We analyzed the growth of MC within the A431 human epithelial cell line and the dependence of this growth on iron uptake. We present evidence here that chelation of iron from infected tissue culture cells with Desferal strongly inhibited intracellular replication of wild-type (wt) MC. We also provide genetic evidence that iron must be acquired by MC from the host cell in order for it to replicate. An hmbR mutant that is unable to use hemoglobin iron and could not grow in tissue culture media without iron supplementation replicated more rapidly within epithelial cells than its wt parent strain. An fbpA mutant that is unable to utilize human transferrin iron or lactoferrin iron replicated normally within cells. In contrast, a tonB mutant could not replicate intracellularly unless infected cultures were supplemented with ferric nitrate. Taken together, these findings strongly suggest that MC intracellular replication requires TonB-dependent uptake of a novel host cell iron source. PMID- 11854235 TI - Transmission intensity affects both antigen-specific and nonspecific T-cell proliferative responses in Loa loa infection. AB - T-cell proliferative responses were studied in two villages in Gabon with different levels of Loa loa transmission. The first village (Okoumbi) had an annual transmission potential (ATP) of approximately 9,000 infective larvae (L3)/person/year (high transmission village), while the second village (Ndjokaye) had an ATP of approximately 1,000 L3/person/year (low transmission village). Proliferation and cytokine assays were performed on peripheral blood mononuclear cells (PBMC) from individuals aged 18 years and over using either mitogens (concanavalin A or phytohemagglutinin), antigens (purified protein derivative [PPD], irrelevant antigen), or soluble extracts of L3, microfilariae, or adult L. loa. PBMC from individuals in the low transmission village responded better to stimulation with adult antigen and to PPD than did PBMC from individuals in the high transmission village (P = 0.0031 and P = 0.0012, respectively). These data suggest that high levels of transmission of L. loa depress both specific and nonspecific T-cell proliferative responses in infected humans. PMID- 11854234 TI - Naturally exposed populations differ in their T1 and T2 responses to the circumsporozoite protein of Plasmodium falciparum. AB - T-cell responses directed against the circumsporozoite protein (CS) of Plasmodium falciparum can mediate protection against malaria. We determined the frequency of T cells reactive to different regions of the CS in the blood of donors naturally exposed to P. falciparum by examining T1 (gamma interferon [IFN-gamma] ELISPOT assay), T2 (interleukin 4 [IL-4] ELISPOT assay), and proliferative T-cell responses. The proliferative responses were weak, which confirmed previous observations. The responses to the CS in the IL-4 and IFN-gamma ELISPOT assays were also weak (<40 responding cells per 10(6) cells), much weaker than the response to the purified protein derivative of Mycobacterium tuberculosis in the same donors. Moreover, a response in one assay could not be used to predict a response in either of the other assays, suggesting that although these assays may measure different responding cells, all of the responses are weakly induced by natural exposure. Interestingly, the two different study populations used had significantly different T1 and T2 biases in their responses in the C terminus of the protein, suggesting that the extent of P. falciparum exposure can affect regulation of the immune system. PMID- 11854236 TI - Expression of surfactant protein D in the human gastric mucosa and during Helicobacter pylori infection. AB - Helicobacter pylori establishes persistent infection of gastric mucosa with diverse clinical outcomes. The innate immune molecule surfactant protein D (SP-D) binds selectively to microorganisms, inducing aggregation and phagocytosis. In this study, we demonstrated the expression of SP-D in gastric mucosa by reverse transcription-PCR and immunohistochemical analysis. SP-D is present at the luminal surface and within the gastric pits, with maximal expression at the surface. Levels of expression are significantly increased in H. pylori-associated gastritis compared to those in the normal mucosa. Immunofluorescence microscopy was used to demonstrate binding and agglutination of H. pylori by SP-D in a lectin-specific manner. These activities resulted in a 50% reduction in the motility of H. pylori, as judged on the basis of curvilinear velocity measured by using a Hobson BacTracker. Lipopolysaccharides extracted from three H. pylori strains were shown to bind SP-D in a concentration-dependent manner, and there was marked variation in the avidity of binding among the strains. SP-D may therefore play a significant role in the innate immune response to H. pylori infection. PMID- 11854237 TI - Modulation of immune responses to Mycobacterium bovis in cattle depleted of WC1(+) gamma delta T cells. AB - It is accepted that cell-mediated immune responses predominate in mycobacterial infections. Many studies have shown that CD4(+) T cells produce Th1 cytokines, such as gamma interferon (IFN-gamma), in response to mycobacterial antigens and that the cytolytic activity of CD8(+) cells toward infected macrophages is important. However, the extent and manner in which gamma delta T cells participate in this response remain unclear. In ruminants, gamma delta T cells comprise a major proportion of the peripheral blood mononuclear cell population. We have previously shown that WC1(+) gamma delta T cells are involved early in Mycobacterium bovis infection of cattle, but their specific functions are not well understood. Here we describe an in vivo model of bovine tuberculosis in which the WC1(+) gamma delta T cells were depleted from the peripheral circulation and respiratory tract, by infusion of WC1(+)-specific monoclonal antibody, prior to infection. While no effects on disease pathology were observed in this experiment, results indicate that WC1(+) gamma delta T cells, which become significantly activated (CD25(+)) in the circulation of control calves from 21 days postinfection, may play a role in modulating the developing immune response to M. bovis. WC1(+)-depleted animals exhibited decreased antigen specific lymphocyte proliferative response, an increased antigen-specific production of interleukin-4, and a lack of specific immunoglobulin G2 antibody. This suggests that WC1(+) gamma delta TCR(+) cells contribute, either directly or indirectly, toward the Th1 bias of the immune response in bovine tuberculosis--a hypothesis supported by the decreased innate production of IFN-gamma, which was observed in WC1(+)-depleted calves. PMID- 11854238 TI - Survival of Tropheryma whipplei, the agent of Whipple's disease, requires phagosome acidification. AB - Tropheryma whipplei was established as the agent of Whipple's disease in 2000, but the mechanisms by which it survives within host cells are still unknown. We show here that T. whipplei survives within HeLa cells by controlling the biogenesis of its phagosome. Indeed, T. whipplei colocalized with lysosome associated membrane protein 1, a membrane marker of late endosomal and lysosomal compartments, but not with cathepsin D, a lysosomal hydrolase. This defect in phagosome maturation is specific to live organisms, since heat-killed bacilli colocalized with cathepsin D. In addition, T. whipplei survived within HeLa cells by adapting to acidic pH. The vacuoles containing T. whipplei were acidic (pH 4.7 +/- 0.3) and acquired vacuolar ATPase, responsible for the acidic pH of late phagosomes. The treatment of HeLa cells with pH-neutralizing reagents, such as ammonium chloride, N-ethylmaleimide, bafilomycin A1, and chloroquine, increased the intravacuolar pH and promoted the killing of T. whipplei. The ability of T. whipplei to survive in an acidic environment and to interfere with phagosome lysosome fusion is likely critical for its prolonged persistence in host cells during the course of Whipple's disease. Our results suggest that manipulating the intravacuolar pH may provide a new approach for the treatment of Whipple's disease. PMID- 11854239 TI - Construction and characterization of a live, attenuated aroA deletion mutant of Pseudomonas aeruginosa as a candidate intranasal vaccine. AB - Antibodies to the lipopolysaccharide O antigen of Pseudomonas aeruginosa mediate high-level immunity, but protective epitopes have proven to be poorly immunogenic, while nonprotective or minimally protective O-antigen epitopes often elicit the best immune responses. With the goal of developing a broadly protective P. aeruginosa vaccine, we used a gene replacement system based on the Flp recombinase to construct an unmarked aroA deletion mutant of the P. aeruginosa serogroup O2/O5 strain PAO1. The resultant aroA deletion mutant of PAO1 is designated PAO1 Delta aroA. The aroA deletion was confirmed by both PCR and failure of the mutant to grow on minimal media lacking aromatic amino acids. When evaluated for safety and immunogenicity in mice, PAO1 Delta aroA could be applied either intranasally or intraperitoneally at doses up to 5 x 10(9) CFU per mouse without adverse effects. No dissemination of PAO1 Delta aroA to blood, liver, or spleen was detected after intranasal application, and histological evidence of pneumonia was minimal. Intranasal immunization of mice and rabbits elicited high titers of immunoglobulin G to whole bacterial cells and to heat stable bacterial antigens of all seven prototypic P. aeruginosa serogroup O2/O5 strains. The mouse antisera mediated potent phagocytic killing of most of the prototypic serogroup O2/O5 strains, while the rabbit antisera mediated phagocytic killing of several serogroup-heterologous strains in addition to killing all O2/O5 strains. This live, attenuated P. aeruginosa strain PAO1 Delta aroA appears to be safe for potential use as an intranasal vaccine and elicits high titers of opsonic antibodies against multiple strains of the P. aeruginosa O2/O5 serogroup. PMID- 11854240 TI - Identification of a Mycobacterium tuberculosis putative classical nitroreductase gene whose expression is coregulated with that of the acr aene within macrophages, in standing versus shaking cultures, and under low oxygen conditions. AB - Tuberculosis remains a leading killer worldwide, and new approaches for its treatment and prevention are urgently needed. This effort will benefit greatly from a better understanding of gene regulation in Mycobacterium tuberculosis, particularly with respect to this pathogen's response to its host environment. We examined the behavior of two promoters from the divergently transcribed M. tuberculosis genes acr/hspX/Rv2031c (alpha-crystallin homolog) and Rv2032/acg (acr-coregulated gene) by using a promoter-GFP fusion assay in Mycobacterium bovis BCG. We found that Rv2032 is a novel macrophage-induced gene whose expression is coregulated with that of acr. Relative levels of intracellular induction for both promoters were significantly affected by shallow standing versus shaking bacterial culture conditions prior to macrophage infection, and both promoters were strongly induced under low oxygen conditions. Deletion analyses showed that DNA sequences within a 43-bp region were required for expression of these promoters under all conditions. Multiple sequence alignment and database searches performed with PROBE indicated that Rv2032 is one of eight M. tuberculosis genes of previously unknown function that belong to an unusual superfamily of classical nitroreductases, which may have a role for bacteria within the host environment. These findings show that mycobacterial culture conditions can greatly influence the results and interpretation of subsequent gene regulation experiments. We propose that these differences might be exploited for dissection of the regulatory factors that affect mycobacterial gene expression within the host. PMID- 11854241 TI - Interaction of an uuter membrane protein of enterotoxigenic Escherichia coli with cell surface heparan sulfate proteoglycans. AB - We have previously shown that enterotoxigenic invasion protein A (Tia), a 25-kDa outer membrane protein encoded on an apparent pathogenicity island of enterotoxigenic Escherichia coli (ETEC) strain H10407, mediates attachment to and invasion into cultured human gastrointestinal epithelial cells. The epithelial cell receptor(s) for Tia has not been identified. Here we show that Tia interacts with cell surface heparan sulfate proteoglycans. Recombinant E. coli expressing Tia mediated invasion into wild-type epithelial cell lines but not invasion into proteoglycan-deficient cells. Furthermore, wild-type eukaryotic cells, but not proteoglycan-deficient eukaryotic cells, attached to immobilized polyhistidine tagged recombinant Tia (rTia). Binding of epithelial cells to immobilized rTia was inhibited by exogenous heparan sulfate glycosaminoglycans but not by hyaluronic acid, dermatan sulfate, or chondroitin sulfate. Similarly, pretreatment of eukaryotic cells with heparinase I, but not pretreatment of eukaryotic cells with chrondroitinase ABC, inhibited attachment to rTia. In addition, we also observed heparin binding to both immobilized rTia and recombinant E. coli expressing Tia. Heparin binding was inhibited by a synthetic peptide representing a surface loop of Tia, as well as by antibodies directed against this peptide. Additional studies indicated that Tia, as a prokaryotic heparin binding protein, may also interact via sulfated proteoglycan molecular bridges with a number of mammalian heparan sulfate binding proteins. These findings suggest that the binding of Tia to host epithelial cells is mediated at least in part through heparan sulfate proteoglycans and that ETEC belongs on the growing list of pathogens that utilize these ubiquitous cell surface molecules as receptors. PMID- 11854242 TI - Chemokine expression patterns differ within anatomically distinct regions of the genital tract during Chlamydia trachomatis infection. AB - Untreated infections with Chlamydia trachomatis commonly result in ascending infection to fallopian tubes and subsequent immune-mediated tubal pathology in females. The proposed immune-mediated injury may be associated with the increased recruitment of CD4 cells to the upper genital tract (GT) (oviducts) in comparison to the lower GT (cervix) during infection, as shown in animal models. To understand the mechanisms responsible for this biased recruitment of CD4 cells within the GT, we characterized chemokine expression patterns in the upper and lower GTs in mice during infection with the murine pneumonitis biovar of Chlamydia trachomatis. Enzyme-linked immunosorbent assays of supernatants from GT homogenates revealed that the levels of the Th1-associated chemokines CXCL9 (monokine induced by gamma interferon), CXCL10 (interferon-inducible protein 10), and CCL5 (RANTES) were significantly higher in the upper GT than in the lower GT after infection, while the CCL3 (macrophage inflammatory protein 1 alpha) level was not increased. In contrast, the level of chemokine CCL11 (eotaxin) was significantly elevated in the lower GT later in the course of infection. Increased levels of mRNA confirmed the selective differences in chemokine expression within the upper and lower GTs. The increased levels of Th1-inducible chemokines in the upper GT were not due to differences in the magnitude of infection or progesterone pretreatment. These data demonstrate that the upper and lower regions of the GT respond differently to Chlamydia infection. PMID- 11854244 TI - Temporal expression of the Candida albicans genes CHK1 and CSSK1, adherence, and morphogenesis in a model of reconstituted human esophageal epithelial candidiasis. AB - We previously demonstrated that genes encoding a putative two-component histidine kinase (CHK1) or a response regulator (CSSK1) are each required for virulence in a murine model of hematogenously disseminated candidiasis and that strains with each gene deleted are also defective in morphogenesis under certain growth conditions. In the present study, the role of these two genes in the adherence to and colonization of reconstituted human esophageal tissue (RHE) is described. We compared strains of Candida albicans with deletions of chk1 (strain CHK21) and cssk1 (strain CSSK21) to wild-type cells (CAF2), as well as strains with CHK1 and CSSK1 reconstituted (strains CHK23 and CSSK23, respectively). Adherence and colonization of RHE were evaluated in periodic acid-Schiff-stained sections, as well as by SEM. We observed that both deletion-containing strains colonized the RHE to a lesser extent than did CAF2 and that the percent germination by both strains was reduced in comparison to that of control strains at 1 h postinfection. Expression of CHK1 or CSSK1 was quantitated by reverse transcription (RT)-PCR from RHE tissues infected with wild-type C. albicans yeast cells. Expression of both CHK1 and CSSK1 increased over the 48-h period following infection of the tissue, although expression of CHK1 was greater than that of CSSK1. By RT-PCR, we have also shown that expression of CHK1 and CSSK1 in the strains with cssk1 and chk1 deleted, respectively, was similar to that of CAF2, indicating that CHK1 and CSSK1 do not regulate each other but probably encode signal proteins of different pathways. Our observations indicate that CHK1 and CSSK1 are each partially required for colonization and conversion to filamentous growth on RHE tissue. PMID- 11854243 TI - Role of inflammatory mediators in resistance and susceptibility to pneumococcal infection. AB - Variations in the host response during pneumonia caused by Streptococcus pneumoniae in susceptible (CBA/Ca) and resistant (BALB/c) inbred mouse strains were investigated. Significant differences were detected in survival time, core body temperature, lung-associated and systemic bacterial loads, mast cell numbers, magnitude and location of cytokine production, lung disruption, and ability of isolated lung cells to release the cytokine tumor necrosis factor (TNF) alpha in vitro. Overall, the results indicate that the reduced capacity of CBA/Ca mice to induce rapid TNF activity within the airways following infection with S. pneumoniae may be a factor in their elevated susceptibility to pneumococcal pneumonia. PMID- 11854245 TI - Naturally attenuated, orally administered Mycobacterium microti as a tuberculosis vaccine is better than subcutaneous Mycobacterium bovis BCG. AB - Mycobacterium microti is phylogenetically closely related to Mycobacterium tuberculosis and is a member of that complex of organisms. It is a curved, acid fast bacillus that is naturally attenuated with a narrow host range for Microtus species only. In this study, we confirm the unique susceptibility of voles to infection with M. microti and the relative resistance of mice with a significantly lower organism burden after 8 weeks of infection. In addition, histopathologic examination of lungs reveals a lack of cellular, granulomatous aggregates characteristically seen in murine M. tuberculosis infection. In the past, M. microti has been used successfully in humans as a vaccine against tuberculosis but was associated with cutaneous reactions. In an attempt to circumvent this adverse effect, we report the efficacy of aerosol and oral vaccination with M. microti. High-dose orogastric vaccination with M. microti resulted in a statistically significant improvement in protection against aerosol challenge with virulent M. tuberculosis in the murine model compared with subcutaneous M. bovis BCG Pasteur vaccination. PMID- 11854246 TI - Antibody-mediated protection in murine Cryptococcus neoformans infection is associated with pleotrophic effects on cytokine and leukocyte responses. AB - Cryptococcus neoformans, an encapsulated yeast, is a common cause of life threatening meningoencephalitis in immunosuppressed patients. We previously observed that administration of a monoclonal antibody (MAb) to the capsular polysaccharide to mice with pulmonary infection prolonged survival and enhanced granulomatous inflammation without reducing lung CFU. To understand the mechanism of MAb action, we studied leukocyte recruitment and cytokine profiles in lungs of A/JCr mice. B lymphocytes were the predominant cell type in lung infiltrates, comprising 15 to 30% of the leukocytes. Despite alterations in histological appearance, fluorescence-activated cell sorter analysis revealed no significant difference in total numbers of lung leukocytes in MAb-treated mice and controls. Differences in the immune response to C. neoformans between MAb-treated mice and controls included (i) an increase in the percentage of granulocytes among lung leukocytes on day 14, (ii) higher macrophage surface expression of CD86 on day 28, (iii) larger amounts of IL-10 in lung homogenates at day 7, (iv) a trend toward smaller amounts of gamma interferon mRNA and protein on day 7, and (v) a smaller increase in the levels of interleukin-4 mRNA and protein on day 7. Hence, the immune responses to C. neoformans infection in the presence and absence of specific antibody were qualitatively similar, and antibody administration was associated with several subtle quantitative differences in immune response parameters that could translate into enhanced survival. MAb may function partly by down-regulating the inflammatory response and reducing host damage. Our findings demonstrate unexpected complexity in the interaction between specific MAb and other components of the host immune response. PMID- 11854247 TI - Monoclonal antibody against the Plasmodium falciparum chitinase, PfCHT1, recognizes a malaria transmission-blocking epitope in Plasmodium gallinaceum ookinetes unrelated to the chitinase PgCHT1. AB - To initiate invasion of the mosquito midgut, Plasmodium ookinetes secrete chitinases that are necessary to cross the chitin-containing peritrophic matrix en route to invading the epithelial cell surface. To investigate chitinases as potential immunological targets of blocking malaria parasite transmission to mosquitoes, a monoclonal antibody (MAb) was identified that neutralized the enzymatic activity of the sole chitinase of Plasmodium falciparum, PfCHT1, identified to date. This MAb, designated 1C3, previously shown to react with an apical structure of P. falciparum ookinetes, also reacts with a discrete apical structure of P. gallinaceum ookinetes. In membrane feeding assays, MAb 1C3 markedly inhibited P. gallinaceum oocyst development in mosquito midguts. MAb 1C3 affinity isolated an approximately 210-kDa antigen which, under reducing conditions, became a 35-kDa antigen. This isolated 35-kDa protein cross-reacted with an antiserum raised against a synthetic peptide derived from the P. gallinaceum chitinase active site, PgCHT1, even though MAb 1C3 did not recognize native or recombinant PgCHT1 on Western blot. Therefore, this affinity-purified 35-kDa antigen appears similar to a previously identified protein, PgCHT2, a putative second chitinase of P. gallinaceum. Epitope mapping indicated MAb 1C3 recognized a region of PfCHT1 that diverges from a homologous amino acid sequence conserved within sequenced chitinases of P. berghei, P. yoelii, and P. gallinaceum (PgCHT1). A synthetic peptide derived from the mapped 1C3 epitope may be useful as a component of a subunit transmission-blocking vaccine. PMID- 11854248 TI - Lack of fusion of azurophil granules with phagosomes during phagocytosis of Mycobacterium smegmatis by human neutrophils is not actively controlled by the bacterium. AB - Biogenesis of phagolysosomes is a very rapid event in neutrophils which takes place with nascent unclosed phagosomes, leading to the release of lysosomal enzymes such as beta-glucuronidase in the extracellular medium. We have previously shown that, under nonopsonic conditions, both pathogenic and nonpathogenic mycobacteria uncouple phagocytosis from fusion of azurophil granules (specialized secretory lysosomes) with phagosomes. In the present study we questioned whether they actively act on neutrophils to block this process or use phagocytic receptors that negatively control the biogenesis of phagolysosomes. As for live unicellular Mycobacterium smegmatis, we observed that nonopsonic phagocytosis of heat-killed mycobacteria did not induce the release of beta-glucuronidase, indicating that M. smegmatis does not actively act on the fusion process in neutrophils. In contrast, phagocytosis of unicellular M. smegmatis opsonized in immune serum or that of small nonopsonized mycobacterial aggregates restored the biogenesis of phagolysosomes. Aggregates were internalized in a CR3- and cholesterol-dependent manner as unicellular mycobacteria. However, aggregates but not unicellular bacteria triggered F-actin and Hck recruitment at the phagosomes, events that have been associated with lysosome fusion. Thus, we propose that M. smegmatis does not actively control the fusion of azurophil granules at early time points postinfection and that mycobacterial aggregates recruit large clusters of receptors at the neutrophil surface which could trap proteins implicated in the biogenesis of phagolysosomes. PMID- 11854249 TI - Babesia bovis merozoite surface antigen 1 and rhoptry-associated protein 1 are expressed in sporozoites, and specific antibodies inhibit sporozoite attachment to erythrocytes. AB - We examined Babesia bovis sporozoites for the expression of two molecules, merozoite surface antigen 1 (MSA-1) and rhoptry-associated protein 1 (RAP-1), that are postulated to be involved in the invasion of host erythrocytes. Both MSA 1 and RAP-1 were transcribed and expressed in infectious sporozoites. Importantly, monospecific MSA-1 and RAP-1 antisera each inhibited sporozoite invasion of erythrocytes in vitro. This is the first identification of antigens expressed in Babesia sp. sporozoites and establishes that, at least in part, sporozoites and merozoites share common targets of antibody mediated inhibition of erythrocyte invasion. PMID- 11854250 TI - Influence of the alternative sigma(28) factor on virulence and flagellum expression of Legionella pneumophila. AB - The fliA gene of Legionella pneumophila encoding the alternative sigma(28) factor was inactivated by introducing a kanamycin resistance cassette. Electron microscopy and Western blot analysis revealed that the fliA mutant strain is aflagellate and expresses no flagellin. Reporter gene assays indicated that the flaA promoter is not active in the fliA mutant strain. The fliA mutant strain multiplied less effectively in coculture with amoebae than the wild-type strain and was not able to replicate in coculture with Dictyostelium discoideum. PMID- 11854251 TI - HLA-B27-transfected (Salmonella permissive) and HLA-A2-transfected (Salmonella nonpermissive) human monocytic U937 cells differ in their production of cytokines. AB - The cytokine secretion of the Salmonella-permissive, HLA-B27-positive U937 cells was examined, as it was previously shown that these cells kill Salmonella less efficiently than controls. Salmonella-permissive U937 cells showed upregulated production of interleukin 10 and to a lesser extent tumor necrosis factor alpha. HLA-B27-associated modulation of cytokine responses may have importance in the pathogenesis of reactive arthritis. PMID- 11854252 TI - Lipopolysaccharide-specific but not anti-flagellar immunoglobulin A monoclonal antibodies prevent Salmonella enterica serotype enteritidis invasion and replication within HEp-2 cell monolayers. AB - The protective potential of immunoglobulin A (IgA) monoclonal antibodies (MAbs) directed against O and H antigens of Salmonella enterica serotype Enteritidis to prevent bacterial adhesion to and invasion of HEp-2 cells was evaluated. Although anti-flagellar IgA MAbs showed strong agglutinating capacities, they did not protect cell monolayers. In contrast, IgA MAbs specific for the O:9 epitope of Salmonella lipopolysaccharide antigen alone prevented S. enterica serotype Enteritidis entry and replication within HEp-2 cells, and the protection was not mediated by direct binding of antibodies to bacterial adhesins or by agglutination of microorganisms. PMID- 11854253 TI - Analyses of the evolutionary distribution of Salmonella translocated effectors. AB - The type III secretion system encoded by Salmonella pathogenicity island 2 (SPI2) translocates Salmonella translocated effectors (STE) into host cells. STE are encoded by genes outside of SPI2. The distribution of STE loci within the salmonellae was investigated. In contrast to the SPI2 locus that is conserved within Salmonella enterica, STE loci show a variable distribution. In addition to other virulence determinants, the possession of various sets of STE loci may contribute to the different host ranges and pathogenic potentials of S. enterica serovars. PMID- 11854254 TI - Protective immunity against Mycobacterium tuberculosis induced by dendritic cells pulsed with both CD8(+)- and CD4(+)-T-cell epitopes from antigen 85A. AB - Immunization with DNA followed by modified vaccinia virus Ankara strain, both expressing the antigen 85A, induced both CD4(+)- and CD8(+)-T-cell responses in BALB/c mice. Following challenge with Mycobacterium tuberculosis, this prime boost regimen produced protection equivalent to that conferred by Mycobacterium bovis BCG. Following immunization with dendritic cells pulsed with an antigen 85A CD4(+)- or CD8(+)-restricted epitope, alone or in combination, copresentation of both epitopes on the same dendritic cell was required for protection, demonstrating that induced CD8(+) T cells can play a protective role against tuberculosis. PMID- 11854255 TI - Increased interleukin 4 (IL-4) receptor expression and IL-4-induced decrease in IL-12 production by Langerhans cells infected with Leishmania major. AB - Langerhans cells (LC) take up Leishmania major and are critical for the induction of the parasite-specific T-cell response. Their functional activities are regulated by cytokines. We analyzed whether infection of LC with L. major modulates the expression of their cytokine receptors. The expression of the interleukin 4 (IL-4) receptor was increased on infected LC from susceptible mice but not on those from resistant mice. Moreover, IL-4 treatment strongly decreased the lipopolysaccharide-induced IL-12 response of infected LC from susceptible mice. This modulation of IL-4 receptor expression and IL-12 production by infection of LC with Leishmania may contribute to the development of Th2 cells and to susceptibility to infection. PMID- 11854256 TI - Induction of dnaK through its native heat shock promoter is necessary for intramacrophagic replication of Brucella suis. AB - The heat shock protein DnaK is essential for intramacrophagic replication of Brucella suis. The replacement of the stress-inducible, native dnaK promoter of B. suis by the promoter of the constitutively expressed bla gene resulted in temperature-independent synthesis of DnaK. In contrast to a dnaK null mutant, this strain grew at 37 degrees C, with a thermal cutoff at 39 degrees C. However, the constitutive dnaK mutant, which showed high sensitivity to H(2)O(2)-mediated stress, failed to multiply in murine macrophage-like cells and was rapidly eliminated in a mouse model of infection, adding strong arguments to our hypothesis that stress-mediated and heat shock promoter-dependent induction of dnaK is a crucial event in the intracellular replication of B. suis. PMID- 11854257 TI - Virulence of Streptococcus pneumoniae: PsaA mutants are hypersensitive to oxidative stress. AB - psaA encodes a 37-kDa pneumococcal lipoprotein which is part of an ABC Mn(II) transport complex. Streptococcus pneumoniae D39 psaA mutants have previously been shown to be significantly less virulent than wild-type D39, but the mechanism underlying the attenuation has not been resolved. In this study, we have shown that psaA and psaD mutants are highly sensitive to oxidative stress, i.e., to superoxide and hydrogen peroxide, which might explain why they are less virulent than the wild-type strain. Our investigations revealed altered expression of the key oxidative-stress response enzymes superoxide dismutase and NADH oxidase in psaA and psaD mutants, suggesting that PsaA and PsaD may play important roles in the regulation of expression of oxidative-stress response enzymes and intracellular redox homeostasis. PMID- 11854259 TI - Selective T-cell recognition of the N-terminal peptide of GroES in tuberculosis. AB - Peptides derived from the whole sequence of mycobacterial GroES heat shock proteins were tested for the ability to induce the proliferation of blood mononuclear cells from tuberculosis patients and sensitized healthy subjects. The response to the N-terminal peptide (residues 1 to 16) was found to be more frequent and stronger in tuberculosis patients. This finding is exceptional, considering that recognition of all other GroES peptides by patients was either diminished or not different from that of controls. PMID- 11854258 TI - Role of cholesterol and the ganglioside GM(1) in entry and short-term survival of Brucella suis in murine macrophages. AB - Brucella species are gram-negative, facultative intracellular bacteria that infect humans and animals. These organisms can survive and replicate within a membrane-bound compartment inside professional and nonprofessional phagocytic cells. Inhibition of phagosome-lysosome fusion has been proposed as a mechanism for intracellular survival in both types of cells. We have previously shown that the maturation inhibition of the Brucella-containing phagosome appears to be restricted at the phagosomal membrane, but the precise molecular mechanisms and factors involved in this inhibition have yet to be identified. Interestingly, recent studies have revealed that caveolae or lipid rafts are implicated in the entry of some microorganisms into host cells and mediate an endocytic pathway avoiding fusion with lysosomes. In this study, we investigated the role of cholesterol and the ganglioside GM(1), two components of lipid rafts, in entry and short-term survival of Brucella suis in murine macrophages, by using cholesterol-sequestering (filipin and beta-methyl cyclodextrin) and GM(1)-binding (cholera toxin B) molecules. Our results suggest that lipid rafts may provide a portal for entry of Brucella into murine macrophages under nonopsonic conditions, thus allowing phagosome-lysosome fusion inhibition, and provide further evidence to support the idea that the phagosome maturation inhibition is restricted at the phagosomal membrane. PMID- 11854260 TI - Discordance between T-cell receptor expression and effector function in mice infected with Histoplasma capsulatum. AB - V beta 10(+) and V beta 14(+) T cells were selectively increased 7 to 14 days following infection in the lungs of naive mice infected with Histoplasma capsulatum. Following secondary challenge of immune mice, V beta 1(+) and V beta 8.1(+) cells were sporadically increased. Elimination of V beta 10(+) and V beta 14(+) cells from naive mice did not alter the course of infection over a period of 21 days. Thus, overexpression of V beta families does not necessarily signify a key role in host defense. PMID- 11854261 TI - A recombinant carboxy-terminal domain of the protective antigen of Bacillus anthracis protects mice against anthrax infection. AB - The immunogenicity and protective efficacy of overlapping regions of the protective antigen (PA) polypeptide, cloned and expressed as glutathione S transferase fusion proteins, have been assessed. Results show that protection can be attributed to individual domains and imply that it is domain 4 which contains the dominant protective epitopes of PA. PMID- 11854264 TI - Genome-wide transcriptional orchestration of circadian rhythms in Drosophila. AB - Circadian rhythms govern the behavior, physiology, and metabolism of living organisms. Recent studies have revealed the role of several genes in the clock mechanism both in Drosophila and in mammals. To study how gene expression is globally regulated by the clock mechanism, we used a high density oligonucleotide probe array (GeneChip) to profile gene expression patterns in Drosophila under light-dark and constant dark conditions. We found 712 genes showing a daily fluctuation in mRNA levels under light-dark conditions, and among these the expression of 115 genes was still cycling in constant darkness, i.e. under free running conditions. Unexpectedly the expression of a large number of genes cycled exclusively under constant darkness. We found that cycling in most of these genes was lost in the arrhythmic Clock (Clk) mutant under light-dark conditions. Expression of periodically regulated genes is coordinated locally on chromosomes where small clusters of genes are regulated jointly. Our findings reveal that many genes involved in diverse functions are under circadian control and reveal the complexity of circadian gene expression in Drosophila. PMID- 11854262 TI - The Dot/Icm type IV secretion system of Legionella pneumophila is essential for the induction of apoptosis in human macrophages. AB - We have previously shown that Legionella pneumophila induces caspase 3-dependent apoptosis in mammalian cells during early stages of infection. In this report, we show that nine L. pneumophila strains with mutations in the dotA, dotDCB, icmT, icmGCD, and icmJB loci are completely defective in the induction of apoptosis, in addition to their severe defects in intracellular replication and pore formation mediated cytotoxicity. Importantly, all nine dot/icm mutants were complemented for all their defective phenotypes with the respective wild-type loci. We show that the role of the Dot/Icm type IV secretion system in the induction of apoptosis is independent of the RtxA toxin, the dot/icm-regulated pore-forming toxin, and the type II secretion system. However, the pore-forming toxin, which is triggered upon entry into the postexponential growth phase, enhances the ability of L. pneumophila to induce apoptosis. Our data provide the first example of the role of a type IV secretion system of a bacterial pathogen in the induction of apoptosis in the host cell. PMID- 11854265 TI - Localization of the secretory granule marker protein chromogranin B in the nucleus. Potential role in transcription control. AB - Chromogranins A (CGA) and B (CGB) are two major Ca(2+) storage proteins of the secretory granules of neuroendocrine cells. Nevertheless, we found in the present study that CGB was also localized in the nucleus. In immunogold electron microscopy using bovine adrenal medullary chromaffin cells, it was found that the number of CGB-labeled gold particles localized per microm(2) of the nucleus was equivalent to 20% that of CGB-labeled gold particles localized per microm(2) of the secretory granules. Considering that CGB is estimated to exist in the 0.1-0.2 mm range in the secretory granules of bovine chromaffin cells, 20% of these amounts to 20-40 microm. In addition, transfection of CGA and CGB into nonneuroendocrine COS-7 and NIH3T3 cells repeatedly indicated the nuclear localization of CGB in addition to its usual localization in the cytoplasm. Moreover, immunoblot and immunogold electron microscopy analyses of neuroendocrine PC12 cells also showed the existence of endogenous CGB in both the cytosol and the nucleus. Nuclear routing of CGB did not appear to depend entirely upon the nuclear localization signal as some of the nuclear localization signal mutant CGB were still targeted to the nucleus. In gene array assay, CGB was shown to either induce or suppress transcription of many genes including those of transcription factors. Of these we have analyzed eight genes, four induced (zinc finger protein, MEF2C, hCRP2, abLIM) and four suppressed (hcKrox, T3-receptor, troponin C, integrin) using the quantitative reverse transcription-PCR method and spectrophotometry to determine the transcription levels of each mRNA. CGB was shown to increase the transcription of zinc finger protein, MEF2C, hCRP2, and abLIM by 2.5-5-fold while suppressing that of hcKrox, T3-receptor, troponin C, and integrin by 60-75%. Given that MEF2C and hcKrox genes are transcription factors, these results pointed to the transcription control role of CGB in the nucleus. PMID- 11854266 TI - Effect of phosphorylation on the structure and fold of transactivation domain of p53. AB - Several phosphorylations are known to occur in the N-terminal transactivation domain of human p53. To explore the structural effects of these phosphorylations, we have chemically synthesized the unphosphorylated p53-(1-39) and its three phosphorylated analogs, phosphorylated at Ser-15, Thr-18, and Ser-20. p53-(1-39) and its Ser-15 and Thr-18 phosphorylated analogs were tested for interaction with p300. The order of binding affinities was similar to that derived from biochemical experiments with the whole protein, indicating functional integrity of the domain. Differences in chemical shifts and coupling constants indicate significant structural changes upon phosphorylations. The single tryptophan in the unphosphorylated domain has an emission maximum and a Stern-Volmer constant that are characteristics of tryptophans situated in protein interiors. The diffusion constant is monomer-like, with an axial ratio of 1:7.5, indicating a significant degree of compaction. Upon phosphorylations, the emission maximum and diffusion constant change significantly toward values that indicate more open conformations. Binding of the hydrophobic probe bis-1-anilino-8 naphthalenesulfonate to the unphosphorylated and one of the phosphorylated domains is also significantly different, suggesting different conformations. We propose that phosphorylations switch the largely folded transactivation domain to more open conformations that interact with transcription factors such as p300/cAMP- responsive element-binding protein-binding protein, leading to enhancement of gene expression. PMID- 11854267 TI - The human prepro-orexin gene regulatory region that activates gene expression in the lateral region and represses it in the medial regions of the hypothalamus. AB - Prepro-orexin is a precursor of the neuropeptides orexin-A and -B, which are localized in the neuronal population of the lateral hypothalamic area (LHA). We wished to elucidate the mechanisms by which the prepro-orexin gene is specifically activated in orexin neurons in the LHA. The 3.2-kb 5'-flanking region of the human prepro-orexin gene is sufficient for the specific expression of an Escherichia coli lacZ reporter gene in orexin neurons. Therefore, we examined a series of reporter constructs harboring this 3.2-kb regulatory region or its deletion in a reporter transgenic mouse assay. There are two phylogenetically conserved regions located 287 bp (orexin regulatory element (OE) 1) and 2.5 kb (OE2) upstream of the transcription initiation site of the human prepro-orexin gene. In transgenic mice, both OE1 and OE2 are necessary for expressing the human prepro-orexin gene in the LHA and for repressing its expression in the medial regions of the hypothalamus. Through serial deletion analysis of OE1, we found that the 57-bp core region of OE1 is critical for its spatial gene regulatory function in vivo. Mutation analysis further demonstrated that without contribution from the OE1 core region, the lacZ reporter is expressed ectopically in the medial regions of the hypothalamus. Thus, OE1 contains crucial cis-acting elements regulating prepro-orexin gene expression specifically in the LHA. PMID- 11854268 TI - Molecular determinants of the mechanism underlying acceleration of the interaction between antithrombin and factor Xa by heparin pentasaccharide. AB - The control of coagulation enzymes by antithrombin is vital for maintenance of normal hemostasis. Antithrombin requires the co-factor, heparin, to efficiently inhibit target proteinases. A specific pentasaccharide sequence (H5) in high affinity heparin induces a conformational change in antithrombin that is particularly important for factor Xa (fXa) inhibition. Thus, synthetic H5 accelerates the interaction between antithrombin and fXa 100-fold as compared with only 2-fold versus thrombin. We built molecular models and identified residues unique to the active site of fXa that we predicted were important for interacting with the reactive center loop of H5-activated antithrombin. To test our predictions, we generated the mutants E37A, E37Q, E39A, E39Q, Q61A, S173A, and F174A in human fXa and examined the rate of association of these mutants with antithrombin in the presence and absence of H5. fXa(Q61A) interacts with antithrombin alone with a nearly normal k(ass); however, we observe only a 4-fold increase in k(ass) in the presence of H5. The x-ray crystal structure of fXa reveals that Gln(61) forms part of the S1' and S3' pocket, suggesting that the P' region of the reactive center loop of antithrombin is crucial for mediating the acceleration in the rate of inhibition of fXa by H5-activated antithrombin. PMID- 11854269 TI - ADAMTS4 cleaves at the aggrecanase site (Glu373-Ala374) and secondarily at the matrix metalloproteinase site (Asn341-Phe342) in the aggrecan interglobular domain. AB - Two major proteolytic cleavages, one at NITEGE(373)/A(374)RGSVI and the other at VDIPEN(341)/F(342)FGVGG, have been shown to occur in vivo within the interglobular domain of aggrecan. The Glu(373)-Ala(374) site is cleaved in vitro by aggrecanase-1 (ADAMTS4) and aggrecanase-2 (ADAMTS5), whereas the other site, at Asn(341)-Phe(342), is efficiently cleaved by matrix metalloproteinases (MMPs) and by cathepsin B at low pH. Accordingly, the presence of the cleavage products globular domain 1 (G1)-NITEGE(373) and G1-VDIPEN(341) in vivo has been widely interpreted as evidence for the specific involvement of ADAMTS enzymes and MMPs/cathepsin B, respectively, in aggrecan proteolysis in situ. We show here, in digests with native human aggrecan, that purified ADAMTS4 cleaves primarily at the Glu(373)-Ala(374) site, but also, albeit slowly and secondarily, at the Asn(341)-Phe(342) site. Cleavage at the Asn(341)-Phe(342) site in these incubations was due to bona fide ADAMTS4 activity (and not a contaminating MMP) because the cleavage was inhibited by TIMP-3 (a potent inhibitor of ADAMTS4), but not by TIMP-1 and TIMP-2, at concentrations that totally blocked MMP-3-mediated cleavage at this site. Digestion of recombinant human G1-G2 (wild-type and cleavage site mutants) confirmed the dual activity of ADAMTS4 and supported the idea that the enzyme cleaves primarily at the Glu(373)-Ala(374) site and secondarily generates G1-VDIPEN(341) by removal of the Phe(342)-Glu(373) peptide from G1-NITEGE(373). These results show that G1-VDIPEN(341) is a product of both MMP and ADAMTS4 activities and challenge the widely held assumption that this product represents a specific indicator of MMP- or cathepsin B-mediated aggrecan degradation. PMID- 11854270 TI - Biochemical and genetic properties of Paenibacillus glycosyl hydrolase having chitosanase activity and discoidin domain. AB - Cells of "Paenibacillus fukuinensis" D2 produced chitosanase into surrounding medium, in the presence of colloidal chitosan or glucosamine. The gene of this enzyme was cloned, sequenced, and subjected to site-directed mutation and deletion analyses. The nucleotide sequence indicated that the chitosanase was composed of 797 amino acids and its molecular weight was 85,610. Unlike conventional family 46 chitosanases, the enzyme has family 8 glycosyl hydrolase catalytic domain, at the amino-terminal side, and discoidin domain at the carboxyl-terminal region. Expression of the cloned gene in Escherichia coli revealed beta-1,4-glucanase function, besides chitosanase activity. Analyses by zymography and immunoblotting suggested that the active enzyme was, after removal of signal peptide, produced from inactive 81-kDa form by proteolysis at the carboxyl-terminal region. Replacements of Glu(115) and Asp(176), highly conserved residues in the family 8 glycosylase region, with Gln and Asn caused simultaneous loss of chitosanase and glucanase activities, suggesting that these residues formed part of the catalytic site. Truncation experiments demonstrated indispensability of an amino-terminal region spanning 425 residues adjacent to the signal peptide. PMID- 11854271 TI - A novel zinc finger protein interacts with receptor-interacting protein (RIP) and inhibits tumor necrosis factor (TNF)- and IL1-induced NF-kappa B activation. AB - Receptor-interacting protein (RIP) is a serine/threonine protein kinase that is critically involved in tumor necrosis factor receptor-1 (TNF-R1)-induced NF-kappa B activation. In a yeast two-hybrid screening for potential RIP-interacting proteins, we identified ZIN (zinc finger protein inhibiting NF-kappa B), a novel protein that specifically interacts with RIP. ZIN contains four RING-like zinc finger domains at the middle and a proline-rich domain at the C terminus. Overexpression of ZIN inhibits RIP-, IKK beta-, TNF-, and IL1-induced NF-kappa B activation in a dose-dependent manner in 293 cells. Domain mapping experiments indicate that the RING-like zinc finger domains of ZIN are required for its interaction with RIP and inhibition of RIP-mediated NF-kappa B activation. Overexpression of ZIN also potentiates RIP- and TNF-induced apoptosis. Moreover, immunofluorescent staining indicates that ZIN is a cytoplasmic protein and that it colocalizes with RIP. Our findings suggest that ZIN is an inhibitor of TNF- and IL1-induced NF-kappa B activation pathways. PMID- 11854272 TI - An easily dissociated 26 S proteasome catalyzes an essential ubiquitin-mediated protein degradation pathway in Trypanosoma brucei. AB - The 26 S proteasome, a complex between the 20 S proteasome and 19 S regulatory units, catalyzes ATP-dependent degradation of unfolded and ubiquitinated proteins in eukaryotes. We have identified previously 20 S and activated 20 S proteasomes in Trypanosoma brucei, but not 26 S proteasome. However, the presence of 26 S proteasome in T. brucei was suggested by the hydrolysis of casein by cell lysate, a process that requires ATP but is inhibited by lactacystin, and the lactacystin sensitive turnover of ubiquitinated proteins in the intact cells. T. brucei cDNAs encoding the six proteasome ATPase homologues (Rpt) were cloned and expressed. Five of the six T. brucei Rpt cDNAs, except for Rpt2, were capable of functionally complementing the corresponding rpt deletion mutants of Saccharomyces cerevisiae. Immunoblots showed the presence in T. brucei lysate of the Rpt proteins, which co-fractionated with the yeast 19 S proteasome complex by gel filtration and localized in the 19 S fraction of a glycerol gradient. All the Rpt and putative 19 S non-ATPase (Rpn) proteins were co-immunoprecipitated from T. brucei lysate by individual anti-Rpt antibodies. Treatment of T. brucei cells with a chemical cross-linker resulted in co-immunoprecipitation of 20 S proteasome with all the Rpt and Rpn proteins that sedimented in a glycerol gradient to the position of 26 S proteasome. These data demonstrate the presence of 26 S proteasome in T. brucei cells, which apparently dissociate into 19 S and 20 S complexes upon cell lysis. RNA interference to block selectively the expression of proteasome 20 S core and Rpt subunits resulted in significant accumulation of ubiquitinated proteins accompanied by cessation of cell growth. Expression of yeast RPT2 gene in T. brucei Rpt2-deficient cells could not rescue the lethal phenotype, thus confirming the incompatibility between the two Rpt2s. The T. brucei 11 S regulator (PA26)-deficient RNA interference cells grew normally, suggesting the dispensability of activated 20 S proteasome in T. brucei. PMID- 11854273 TI - Characterization of mouse cysteinyl leukotriene receptors mCysLT1 and mCysLT2: differential pharmacological properties and tissue distribution. AB - Cysteinyl leukotrienes (LTs) are important proinflammatory mediators. Their precise roles in mice need to be elucidated to interpret mouse models of inflammatory diseases. For this purpose, we cloned and characterized mouse receptors for cysteinyl LTs, mCysLT(1) and mCysLT(2). mCysLT(1) and mCysLT(2) were composed of 339 amino acids with 87.3% identity and 309 amino acids with 73.4% identity to human orthologues, respectively. A pharmacological difference was noted between mouse and human CysLT(2). Pranlukast, a specific inhibitor for human CysLT(1), antagonized mCysLT(2) responses as determined by Ca(2+) elevation and receptor-induced promoter activation. The mRNA expressions of both mCysLTs were higher in C57BL/6 mice than in 129 mice. mCysLT(1) mRNA was expressed mainly in skin, lung, and small intestine. mCysLT(2) was seen more ubiquitously with high expressions in spleen, lung, and small intestine. By in situ hybridization we demonstrated for the first time that mCysLT(1) and mCysLT(2) were expressed in subcutaneous fibroblasts. The different pharmacological characteristics of CysLT(2) between human and mouse and the different distributions of CysLTs between mouse strains suggest that careful choice and interpretation are necessary for a study of CysLTs using animal models. PMID- 11854274 TI - Cross-talk in the A1-ATPase from Methanosarcina mazei Go1 due to nucleotide binding. AB - Changes in the A(3)B(3)CDF-complex of the Methanosarcina mazei Go1 A(1)-ATPase in response to ligand binding have been studied by small-angle x-ray scattering, protease digestion, fluorescence spectroscopy, matrix-assisted laser desorption ionization time-of-flight mass spectrometry, and CuCl(2)-induced disulfide formation. The value of the radius of gyration, R(g), increases slightly when MgATP, MgADP, or MgADP + P(i) (but not MgAMP-PNP) is present. The nucleotide binding subunits A and B were reacted with N-4[4-[7-(dimethylamino)-4 methyl]coumarin-3-yl]maleimide, and spectral shifts and changes in fluorescence intensity were detected upon addition of MgAMP-PNP, MgATP, MgADP + P(i), or MgADP. Trypsin treatment of A(1) resulted in cleavage of the stalk subunits C and F, which was rapid in the presence of MgAMP-PNP but slow when MgATP or MgADP were added to the enzyme. When A(1) was supplemented with CuCl(2) a clear nucleotide dependence of an A-A-D cross-linking product was generated in the presence of MgADP and MgATP but not when MgAMP-PNP or MgADP + P(i) was added. The site of cross-link formation was located in the region of the N and C termini of subunit D. The data suggest that the stalk subunits C, D, and F in A(1) undergo conformational changes during ATP hydrolysis. PMID- 11854275 TI - Complementary combining site contact residue mutations of the anti-digoxin Fab 26 10 permit high affinity wild-type binding. AB - Antibody 26-10, obtained in a secondary immune response, binds digoxin with high affinity (K(a) = 1.3 x 10(10) M(-1)) because of extensive shape complementarity. We demonstrated previously that mutations of the hapten contact residue HTrp-100 to Arg (where H refers to the heavy chain) resulted in increased specificity for digoxin analogs substituted at the cardenolide 16 position. However, mutagenesis of H:CDR1 did not result in such a specificity change despite the proximity of the H:CDR1 hapten contact residue Asn-35 to the cardenolide 16 position. Here we constructed a bacteriophage-displayed library containing randomized mutations at H chain residues 30-35 in a 26-10 mutant containing Arg-100 (26-10-RRALD). Phage were selected by panning against digoxin, gitoxin (16-OH), and 16-acetylgitoxin coupled to bovine serum albumin. Clones that retained wild-type Asn at position 35 showed preferred binding to gitoxin, like the 26-10-RRALD parent. In contrast, clones containing Val-35 selected mainly on digoxin-bovine serum albumin demonstrated a shift back to wild-type specificity. Several clones containing Val 35 bound digoxin with increased affinity, approaching that of the wild type in a few instances, in contrast to the mutation Val-35 in the wild-type 26-10 background, which reduces affinity for digoxin 90-fold. It has therefore proven possible to reorder the 26-10 binding site by mutations including two major contact residues on opposite sides of the site and yet to retain high affinity for binding for digoxin. Thus, even among antibodies that have undergone affinity maturation in vivo, different structural solutions to high affinity binding may be revealed. PMID- 11854276 TI - Role of loop structures of neuropsin in the activity of serine protease and regulated secretion. AB - Neuropsin involved in neural plasticity in adult mouse brain is a member of the S1 (clan SA) family of serine proteases and forms characteristic surface loops surrounding the substrate-binding site (Kishi, T., Kato, M., Shimizu, T., Kato, K., Matsumoto, K., Yoshida, S., Shiosaka, S., and Hakoshima, T. (1999) J. Biol. Chem. 274, 4220-4224). Little, however, is known about the roles of these loops. Thus, the present study investigated whether surface loop structures of neuropsin were essential for the generation of enzymatic activity and/or secretion of the enzyme via a regulated secretory pathway. The loops include those stabilized by six disulfide bonds or a loop C (Gly(69)-Glu(80)) and an N-glycosylated kallikrein loop (His(91)-Ile(103)) not containing a site linked by a disulfide bond. First, among the six disulfide bonds, only SS1 in loop E (Gly(142) Leu(155)) and SS6 in loop G (Ser(185)-Gly(197)) were necessary for the catalytic efficiency of neuropsin. Second, disruptions of loop C and the N-linked oligosaccharide chain on the kallikrein loop affected the catalytic efficiency and P2 specificity, respectively. Alternatively, disruptions of loop C and the kallikrein loop enhanced the regulated secretion, whereas there was no one disruption that inhibited the secretion, indicating that there was no critical loop required for the regulated secretion among loops surrounding the substrate binding site. PMID- 11854277 TI - A novel allelic variant of the human TSG-6 gene encoding an amino acid difference in the CUB module. Chromosomal localization, frequency analysis, modeling, and expression. AB - Tumor necrosis factor-stimulated gene-6 (TSG-6) encodes a 35-kDa protein, which is comprised of contiguous Link and CUB modules. TSG-6 protein has been detected in the articular joints of osteoarthritis (OA) patients, with little or no constitutive expression in normal adult tissues. It interacts with components of cartilage matrix (e.g. hyaluronan and aggrecan) and thus may be involved in extracellular remodeling during joint disease. In addition, TSG-6 has been found to have anti-inflammatory properties in models of acute and chronic inflammation. Here we have mapped the human TSG-6 gene to 2q23.3, a region of chromosome 2 linked with OA. A single nucleotide polymorphism was identified that involves a non-synonymous G --> A transition at nucleotide 431 of the TSG-6 coding sequence, resulting in an Arg to Gln alteration in the CUB module (at residue 144 in the preprotein). Molecular modeling of the CUB domain indicated that this amino acid change might lead to functional differences. Typing of 400 OA cases and 400 controls revealed that the A(431) variant identified here is the major TSG-6 allele in Caucasians (with over 75% being A(431) homozygotes) but that this polymorphism is not a marker for OA susceptibility in the patients we have studied. Expression of the Arg(144) and Gln(144) allotypes in Drosophila Schneider 2 cells, and functional characterization, showed that there were no significant differences in the ability of these full-length proteins to bind hyaluronan or form a stable complex with inter-alpha-inhibitor. PMID- 11854278 TI - The chain length dependence of helix formation of the second transmembrane domain of a G protein-coupled receptor of Saccharomyces cerevisiae. AB - The chain length dependence of helix formation of transmembrane peptides in lipids was investigated using fragments corresponding to the second transmembrane domain of the alpha-factor receptor from Saccharomyces cerevisiae. Seven peptides with chain lengths of 10 (M2-10; FKYLLSNYSS), 14 (M2-14), 18 (M2-18), 22 (M2-22), 26 (M2-26), 30 (M2-30) and 35 (M2-35; RSRKTPIFIINQVSLFLIILHSALYFKYLLSNYSS) residues, respectively, were synthesized. CD spectra revealed that M2-10 was disordered, and all of the other peptides assumed partially alpha-helical secondary structures in 99% trifluoroethanol (TFE)/H(2)O. In 50% TFE/H(2)O, M2-30 assumed a beta-like structure. The other six peptides exhibited the same CD patterns as those found in 99% TFE/H(2)O. In 1,2-dimyristoyl-sn-glycero-3 phosphocholine/1,2-dimyristoyl-sn-glycero-3-phospho-rac-(1-glycerol) (4:1 ratio) vesicles, M2-22, M2-26, and M2-35 formed alpha-helical structures, whereas the other peptides formed beta-like structures. Fourier transform infrared spectroscopy in 1,2-dimyristoyl-sn-glycero-3-phosphocholine/1,2-dimyristoyl-sn glycero-3-phospho-rac-(1-glycerol) (4:1) multilayers showed that M2-10, M2-14, M2 18, and M2-30 assumed beta-structures in this environment. Another homologous 30 residue peptide (M2-30B), missing residues SNYSS from the N terminus and extending to RSRKT on the C terminus, was helical in lipid bilayers, suggesting that residues at the termini of transmembrane domains influence their biophysical properties. Attenuated total reflection Fourier transform infrared spectroscopy revealed that M2-22, M2-26, M2-30B, and M2-35 were alpha-helical and oriented at angles of 12 degrees, 13 degrees, 36 degrees, and 34 degrees, respectively, with respect to the multilayer normal. This study showed that chain length must be taken into consideration when using peptides representing single transmembrane domains as surrogates for regions of an intact receptor. Furthermore, this work indicates that the tilt angle and conformation of transmembrane portions of G protein-coupled receptors may be estimated by detailed spectroscopic measurements of single transmembrane peptides. PMID- 11854279 TI - Lipopolysaccharide activates the expression of ISG15-specific protease UBP43 via interferon regulatory factor 3. AB - UBP43 is a protease that specifically removes a ubiquitin-like protein, ISG15, from its targets. Highest levels of UBP43 expression are detected in macrophages and in cell lines of monocytic lineage. Macrophages are important in host defense against bacterial and viral infections. The lipopolysaccharide (LPS) of the bacterial cell wall can mimic bacteria and activate monocytes/macrophages to provoke inflammatory responses. Here, we report that LPS strongly activates UBP43 expression in macrophages, which is paralleled by changes in UBP43 protein levels. Two interferon regulatory factor (IRF) binding sites in the UBP43 promoter are responsible for the induction of UBP43 expression by LPS, as well as for basal UBP43 promoter activity. We have identified two members of the IRF family (IRF-2 and IRF-3) that specifically bind to these sites. IRF-3 plays a primary role in the LPS-inducible activation of the UBP43 gene and IRF-2 confers a basal transcriptional activity to the UBP43 promoter. Furthermore, we demonstrate that LPS treatment increases the amount of ISG15-conjugates in macrophages. Coordinated induction of ISG15 and UBP43 suggests that ISG15 conjugation is a dynamic process and that a critical balance of ISG15 modification should be maintained during innate immune response. PMID- 11854280 TI - The signal peptide of the G protein-coupled human endothelin B receptor is necessary for translocation of the N-terminal tail across the endoplasmic reticulum membrane. AB - The initial step of the intracellular transport of G protein-coupled receptors, their insertion into the membrane of the endoplasmic reticulum, follows one of two different pathways. Whereas one group uses the first transmembrane domain of the mature receptor as an uncleaved signal anchor sequence for this process, a second group possesses additional cleavable signal peptides. The reason this second subset requires the additional signal peptide is not known. Here we have assessed the functional significance of the signal peptide of the endothelin B (ET(B)) receptor in transiently transfected COS.M6 cells. A green fluorescent protein-tagged ET(B) receptor mutant lacking the signal peptide was nonfunctional and retained in the endoplasmic reticulum, suggesting that it has a folding defect. To determine the defect in more detail, ET(B) receptor fragments containing the N-terminal tail, first transmembrane domain, and first cytoplasmic loop were constructed. We assessed N tail translocation across the endoplasmic reticulum membrane in the presence and absence of a signal peptide and show that the signal peptide is necessary for N tail translocation. We postulate that signal peptides are necessary for those G protein-coupled receptors for which post-translational translocation of the N terminus is impaired or blocked by the presence of stably folded domains. PMID- 11854281 TI - Enzyme-substrate interactions in the purine-specific nucleoside hydrolase from Trypanosoma vivax. AB - Nucleoside hydrolases are key enzymes in the purine salvage pathway of Trypanosomatidae and are considered as targets for drug design. We previously reported the first x-ray structure of an inosine-adenosine-guanosine preferring nucleoside hydrolase (IAG-NH) from Trypanosoma vivax (). Here we report the 2.0-A crystal structure of the slow D10A mutant in complex with the inhibitor 3-deaza adenosine and the 1.6-A crystal structure of the same enzyme in complex with a genuine substrate inosine. The enzyme-substrate complex shows the substrate bound to the enzyme in a different conformation from 3-deaza-adenosine and provides a snapshot along the reaction coordinate of the enzyme-catalyzed reaction. The chemical groups on the substrate important for binding and catalysis are mapped. The 2'-OH, 3'-OH, and 5'-OH contribute 4.6, 7.5, and 5.4 kcal/mol to k(cat)/K(m), respectively. Specific interactions with the exocyclic groups on the purine ring are not required for catalysis. Site-directed mutagenesis indicates that the purine specificity of the IAG-NHs is imposed by a parallel aromatic stacking interaction involving Trp(83) and Trp(260). The pH profiles of k(cat) and k(cat)/K(m) indicate the existence of one or more proton donors, possibly involved in leaving group activation. However, mutagenesis of the active site residues around the nucleoside base and an alanine scan of a flexible loop near the active site fail to identify this general acid. The parallel aromatic stacking seems to provide the most likely alternative mechanism for leaving group activation. PMID- 11854282 TI - Positive and negative effects of the major mammalian messenger ribonucleoprotein p50 on binding of 40 S ribosomal subunits to the initiation codon of beta-globin mRNA. AB - p50, the major core protein bound to mammalian mRNAs, has been reported to stimulate translation at low p50/mRNA ratios and inhibit translation at high p50/mRNA ratios. This study aims to address the molecular mechanisms underlying these phenomena using the in vitro assembly of 48 S preinitiation complexes from fully purified translational components in the presence or absence of p50 as analyzed by the toeprint assay. With limited concentrations of eIF2, eIF3, and eIF4F, p50 (but not pyrimidine tract-binding protein, which was taken for comparison) strongly stimulates formation of the 48 S preinitiation complexes with beta-globin mRNA. This stimulation is observed when just a few molecules of p50 are bound per molecule of the mRNA. When the amount of p50 in solution is increased over some threshold p50/mRNA ratio, a remarkable repression is observed that can still be relieved by adding more eIF2 and eIF4F. At even higher concentrations of p50, the inhibitory effect becomes irreversible. The threshold ratio depends upon the extent of secondary structure of the 5'-untranslated region linked to the beta-globin coding region. Chemical probing has confirmed that the binding of p50 to mRNA involves only the sugar-phosphate backbone of the mRNA leaving nucleotide bases free for interaction with other messenger ribonucleoprotein (mRNP) components. These data are best compatible with the functional role of p50 as a "manager" of mRNA-protein interactions in mammalian mRNPs. PMID- 11854283 TI - Respective roles of calcitonin receptor-like receptor (CRLR) and receptor activity-modifying proteins (RAMP) in cell surface expression of CRLR/RAMP heterodimeric receptors. AB - Receptor activity modifying proteins RAMP1, RAMP2, and RAMP3 are responsible for defining affinity to ligands of the calcitonin receptor-like receptor (CRLR). It has also been proposed that receptor activity-modifying proteins (RAMP) are molecular chaperones required for CRLR transport to the cell surface. Here, we have studied the respective roles of CRLR and RAMP in transporting CRLR/RAMP heterodimers to the plasma membrane by using a highly specific binding assay that allows quantitative detection of cell surface-expressed CRLR or RAMP in the Xenopus oocytes expression system. We show that: (i) heterodimer assembly is not a prerequisite for efficient cell surface expression of CRLR, (ii) N-glycosylated RAMP2 and RAMP3 are expressed at the cell surface and their transport to the plasma membrane requires N-glycans, (iii) RAMP1 is not N-glycosylated and is transported to the plasma membrane only upon formation of heterodimers with CRLR, and (iv) introduction of N-glycosylation sites in the RAMP1 sequence (D58N/G60S, Y71N, and K103N/P105S) allows cell surface expression of these mutants at levels similar to that of wild-type RAMP1 co-expressed with CRLR. Our data argue against a chaperone function for RAMP and identify the role of N-glycosylation in targeting these molecules to the cell surface. PMID- 11854284 TI - Decreased metallation and activity in subsets of mutant superoxide dismutases associated with familial amyotrophic lateral sclerosis. AB - Over 90 different mutations in the gene encoding copper/zinc superoxide dismutase (SOD1) cause approximately 2% of amyotrophic lateral sclerosis (ALS) cases by an unknown mechanism. We engineered 14 different human ALS-related SOD1 mutants and obtained high yields of biologically metallated proteins from an Sf21 insect cell expression system. Both the wild type and mutant "as isolated" SOD1 variants were deficient in copper and were heterogeneous by native gel electrophoresis. By contrast, although three mutant SOD1s with substitutions near the metal binding sites (H46R, G85R, and D124V) were severely deficient in both copper and zinc ions, zinc deficiency was not a consistent feature shared by the as isolated mutants. Eight mutants (A4V, L38V, G41S, G72S, D76Y, D90A, G93A, and E133 Delta) exhibited normal SOD activity over pH 5.5-10.5, per equivalent of copper, consistent with the presumption that bound copper was in the proper metal-binding site and was fully active. The H48Q variant contained a high copper content yet was 100-fold less active than the wild type enzyme and exhibited a blue shift in the visible absorbance peak of bound Cu(II), indicating rearrangement of the Cu(II) coordination geometry. Further characterization of these as-isolated SOD1 proteins may provide new insights regarding mutant SOD1 enzyme toxicity in ALS. PMID- 11854285 TI - Familial amyotrophic lateral sclerosis-associated mutations decrease the thermal stability of distinctly metallated species of human copper/zinc superoxide dismutase. AB - We report the thermal stability of wild type (WT) and 14 different variants of human copper/zinc superoxide dismutase (SOD1) associated with familial amyotrophic lateral sclerosis (FALS). Multiple endothermic unfolding transitions were observed by differential scanning calorimetry for partially metallated SOD1 enzymes isolated from a baculovirus system. We correlated the metal ion contents of SOD1 variants with the occurrence of distinct melting transitions. Altered thermal stability upon reduction of copper with dithionite identified transitions resulting from the unfolding of copper-containing SOD1 species. We demonstrated that copper or zinc binding to a subset of "WT-like" FALS mutants (A4V, L38V, G41S, G72S, D76Y, D90A, G93A, and E133Delta) conferred a similar degree of incremental stabilization as did metal ion binding to WT SOD1. However, these mutants were all destabilized by approximately 1-6 degrees C compared with the corresponding WT SOD1 species. Most of the "metal binding region" FALS mutants (H46R, G85R, D124V, D125H, and S134N) exhibited transitions that probably resulted from unfolding of metal-free species at approximately 4-12 degrees C below the observed melting of the least stable WT species. We conclude that decreased conformational stability shared by all of these mutant SOD1s may contribute to SOD1 toxicity in FALS. PMID- 11854286 TI - Isolation and partial characterization of the inactive and active forms of human plasma phospholipid transfer protein (PLTP). AB - Plasma phospholipid transfer protein (PLTP) plays an important role in lipoprotein metabolism. Two forms of PLTP exist in human plasma, one catalytically active (high activity form, HA-PLTP) and the other inactive (low activity form, LA-PLTP) (Oka, T., Kujiraoka, T., Ito, M., Egashira, T., Takahashi, S., Nanjee, N. M., Miller, N. E., Metso, J., Olkkonen, V. M., Ehnholm, C., Jauhiainen, M., and Hattori, H. (2000) J. Lipid Res. 41, 1651-1657). The two forms are associated with macromolecular complexes of different size. The apparent size of LA-PLTP is 520 kDa and that of HA-PLTP is 160 kDa. Of the circulating PLTP mass only a minor portion is in the HA-PLTP form in normolipidemic subjects. In the present study we have isolated and partially characterized the LA and HA forms of PLTP. Both LA- and HA-PLTP bind to heparin Sepharose and can be separated by elution with 0-0.5 m NaCl gradient, with HA PLTP displaying higher affinity for the matrix. LA-PLTP was further purified using hydrophobic butyl-Sepharose and anti-PLTP immunoaffinity chromatography steps. HA-PLTP was subjected to a second heparin-Sepharose step and hydroxylapatite chromatography. Analysis of the two forms of PLTP by SDS-PAGE, Western blotting, immunoprecipitation, and gel filtration demonstrates that LA PLTP is complexed with apoA-I whereas HA-PLTP is not. Instead, HA-PLTP copurified with apoE. Based on these findings we suggest a model in which nascent PLTP enters the circulation as a high specific activity form not associated with apoA I. During or after the transfer of lipolytic surface remnants to HDL, PLTP is transferred to apoA-I-containing HDL particles and thereby becomes part of the low activity complex. PMID- 11854287 TI - Phosphoinositide 3-kinase activity is required for retinoic acid-induced expression and activation of the tissue transglutaminase. AB - Tissue transglutaminase (TGase) is a dual function enzyme that couples an ability to bind GTP with transamidation activity. Retinoic acid (RA) consistently induces TGase expression and activation, and it was recently shown that increased TGase expression protected cells from apoptosis. To better understand how RA regulates TGase, we considered whether RA employed pro-survival signaling pathways to mediate TGase expression and activation. It was found that RA stimulation of NIH3T3 cells activated ERK and phosphoinositide 3-kinase (PI3K); however, only PI3K activation was necessary for RA-induced TGase expression. The overexpression of a constitutively active form of PI3K did not induce TGase expression, indicating that PI3K signaling was necessary but not sufficient for TGase expression. The exposure of cells expressing exogenous TGase to the PI3K inhibitor, LY294002, reduced the ability of TGase to be photoaffinity-labeled with [alpha-(32)P]GTP, providing evidence that PI3K regulates the GTP binding activity of TGase as well as its expression. Moreover, cell viability assays showed that incubation of RA-treated cells with LY294002 together with the TGase inhibitor, monodansylcadaverine (MDC), converted RA from a differentiation factor to an apoptotic stimulus. These findings demonstrate that PI3K activity is required for the RA-stimulated expression and GTP binding activity of TGase, thereby linking the up-regulation of TGase with a well established cell survival factor. PMID- 11854288 TI - The telomeric poly(ADP-ribose) polymerase, tankyrase 1, contains multiple binding sites for telomeric repeat binding factor 1 (TRF1) and a novel acceptor, 182-kDa tankyrase-binding protein (TAB182). AB - Tankyrase 1, a human telomeric poly(ADP-ribose) polymerase, was originally identified through its interaction with TRF1, a negative regulator of telomere length. Tankyrase 1 ADP-ribosylates TRF1 in vitro, and its overexpression induces telomere elongation in human cancer cells. In addition to its telomeric localization, tankyrase 1 resides at multiple subcellular sites, suggesting additional functions for this protein. Here we identify TAB182, a novel tankyrase 1-binding protein of 182 kDa. TAB182 displays a complex pattern of subcellular localization. TAB182 localizes to the nucleus in a heterochromatic staining pattern and to the cytoplasm, where it co-stains with the cortical actin network. TAB182 coimmunoprecipitates with tankyrase 1 from human cells and serves as an acceptor of poly(ADP-ribosyl)ation by tankyrase 1 in vitro. Like TRF1, TAB182 binds to the ankyrin domain (comprising 24 ankyrin repeats) of tankyrase 1. Surprisingly, dissection of this domain reveals multiple discrete and overlapping binding sites for TRF1 and TAB182. Thus, we demonstrate five well conserved ankyrin repeat clusters in tankyrase 1. Although each of the five ankyrin repeat clusters independently binds to TRF1, only three of the five bind toTAB182. These findings suggest that tankyrase 1 may act as a scaffold for large molecular mass complexes made up of multiple binding proteins. We discuss potential roles for tankyrase 1-mediated higher order complexes at telomeres and at other subcellular sites. PMID- 11854289 TI - Hybrid tetramers of porcine liver fructose-1,6-bisphosphatase reveal multiple pathways of allosteric inhibition. AB - Fructose-1,6-bisphosphatase is a square planar tetramer of identical subunits, which exhibits cooperative allosteric inhibition of catalysis by AMP. Protocols for in vitro subunit exchange provide three of five possible hybrid tetramers of fructose-1,6-bisphosphatase in high purity. The two hybrid types with different subunits in the top and bottom halves of the tetramer co-purify. Hybrid tetramers, formed from subunits unable to bind AMP and subunits with wild-type properties, differ from the wild-type enzyme only in regard to their properties of AMP inhibition. Hybrid tetramers exhibit cooperative, potent, and complete (100%) AMP inhibition if at least one functional AMP binding site exists in the top and bottom halves of the tetramer. Furthermore, titrations of hybrid tetramers with AMP, monitored by a tryptophan reporter group, reveal cooperativity and fluorescence changes consistent with an R- to T-state transition, provided that again at least one functional AMP site exists in the top and bottom halves of the tetramer. In contrast, hybrid tetramers, which have functional AMP binding sites in only one half (top/bottom), exhibit an R- to T state transition and complete AMP inhibition, but without cooperativity. Evidently, two pathways of allosteric inhibition of fructose-1,6-bisphosphatase are possible, only one of which is cooperative. PMID- 11854290 TI - Complete inhibition of Streptococcus pneumoniae RecA protein-catalyzed ATP hydrolysis by single-stranded DNA-binding protein (SSB protein): implications for the mechanism of SSB protein-stimulated DNA strand exchange. AB - The ATP-dependent three-strand exchange activity of the Streptococcus pneumoniae RecA protein (RecA(Sp)), like that of the Escherichia coli RecA protein (RecA(Ec)), is strongly stimulated by the single-stranded DNA-binding protein (SSB) from either E. coli (SSB(Ec)) or S. pneumoniae (SSB(Sp)). The RecA(Sp) protein differs from the RecA(Ec) protein, however, in that its ssDNA-dependent ATP hydrolysis activity is completely inhibited by SSB(Ec) or SSB(Sp) protein, apparently because these proteins displace RecA(Sp) protein from ssDNA. These results indicate that in contrast to the mechanism that has been established for the RecA(Ec) protein, SSB protein does not stimulate the RecA(Sp) protein promoted strand exchange reaction by facilitating the formation of a presynaptic complex between the RecA(Sp) protein and the ssDNA substrate. In addition to acting presynaptically, however, it has been proposed that SSB(Ec) protein also stimulates the RecA(Ec) protein strand exchange reaction postsynaptically, by binding to the displaced single strand that is generated when the ssDNA substrate invades the homologous linear dsDNA. In the RecA(Sp) protein-promoted reaction, the stimulatory effect of SSB protein may be due entirely to this postsynaptic mechanism. The competing displacement of RecA(Sp) protein from the ssDNA substrate by SSB protein, however, appears to limit the efficiency of the strand exchange reaction (especially at high SSB protein concentrations or when SSB protein is added to the ssDNA before RecA(Sp) protein) relative to that observed under the same conditions with the RecA(Ec) protein. PMID- 11854291 TI - Generality of the branched pathway in transcription initiation by Escherichia coli RNA polymerase. AB - Transcription initiation has been assumed to be a multi-step sequential process, although additional steps could exist. Initiation from the T7A1 promoter, in particular, apparently behaves in vitro in a manner that can be fully explained by the sequential pathway. However, initiation from the lambda P(R)AL promoter has been shown to follow a branched pathway from which a part of the enzyme promoter complex is arrested at the promoter raising the question as to which mechanism is general. We found that a moribund complex, characteristic of the arrested branch, is formed at the T7A1 promoter, especially in low salt condition indicating that the initiation mechanism for this promoter is also branched. The results of DNA footprinting suggested that holoenzyme in the moribund complex is dislocated on DNA from the position of productive complex. However, only a small fraction of the binary complex becomes arrested at this promoter, and the interconversion between subspecies of binary complex is apparently more reversible than at the lambda P(R)AL promoter, which explains why the reaction pathway appears to be sequential. These findings suggest a generality of the branched pathway mechanism, which would resolve contradictory observations that have been reported for various promoters. PMID- 11854292 TI - Modulation of dimer stability in yeast pyrophosphatase by mutations at the subunit interface and ligand binding to the active site. AB - Yeast (Saccharomyces cerevisiae) pyrophosphatase (Y-PPase) is a tight homodimer with two active sites separated in space from the subunit interface. The present study addresses the effects of mutation of four amino acid residues at the subunit interface on dimer stability and catalytic activity. The W52S variant of Y-PPase is monomeric up to an enzyme concentration of 300 microm, whereas R51S, H87T, and W279S variants produce monomer only in dilute solutions at pH > or = 8.5, as revealed by sedimentation, gel electrophoresis, and activity measurements. Monomeric Y-PPase is considerably more sensitive to the SH reagents N-ethylmaleimide and p-hydroxymercurobenzosulfonate than the dimeric protein. Additionally, replacement of a single cysteine residue (Cys(83)), which is not part of the subunit interface or active site, with Ser resulted in insensitivity of the monomer to SH reagents and stabilization against spontaneous inactivation during storage. Active site ligands (Mg(2+) cofactor, P(i) product, and the PP(i) analog imidodiphosphate) stabilized the W279S dimer versus monomer predominantly by decreasing the rate of dimer to monomer conversion. The monomeric protein exhibited a markedly increased (5-9-fold) Michaelis constant, whereas k(cat) remained virtually unchanged, compared with dimer. These results indicate that dimerization of Y-PPase improves its substrate binding performance and, conversely, that active site adjustment through cofactor, product, or substrate binding strengthens intersubunit interactions. Both effects appear to be mediated by a conformational change involving the C-terminal segment that generally shields the Cys(83) residue in the dimer. PMID- 11854293 TI - Selection of multipotent stem cells during morphogenesis of small intestinal crypts of Lieberkuhn is perturbed by stimulation of Lef-1/beta-catenin signaling. AB - Studies of chimeric mice have disclosed that the stem cell hierarchy in the small intestinal epithelium is established during formation of its proliferative units (crypts of Lieberkuhn). This process involves a selection among several multipotential progenitors so that ultimately only one survives to supply descendants to the fully formed crypt. In this report, we examine the hypothesis that the level of beta-catenin (beta-cat)-mediated signaling is an important factor regulating this stem cell selection. In the canonical Wnt signaling pathway, beta-catenin can partner with Lef-1/Tcf high mobility group (HMG) box transcription factors to control gene expression. Both Lef-1 and Tcf-4 mRNAs are produced in the fetal mouse small intestine. Tcf-4 expression is sustained, whereas Lef-1 levels fall as crypt formation is completed during the first two postnatal weeks. A Tcf-4 gene knockout is known to block intestinal epithelial proliferation in late fetal life. Therefore, to test the hypothesis, we enhanced beta-catenin signaling in a chimeric mouse model in which the stem cell selection could be monitored. A fusion protein containing the HMG box domain of Lef-1 linked to the trans-activation domain of beta-catenin (Lef-1/beta-cat) was constructed to promote direct stimulation of signaling without being retained in the cytoplasm through interactions with E-cadherin and Apc/Axin. Lef-1/beta-cat was expressed in 129/Sv embryonic stem cell-derived small intestinal epithelial progenitors present in developing B6-ROSA26<-->129/Sv chimeras. Lef-1/beta-cat stimulated expression of a known beta-catenin target (E-cadherin), suppressed expression of Apc and Axin, and induced apoptosis in 129/Sv but not in neighboring B6-ROSA26 epithelial cells. This apoptotic response was not associated with any detectable changes in cell division within the Lef-1/beta-cat expressing epithelium. By the time crypt development was completed, all 129/Sv epithelial cells were lost. These results indicate that developmental changes in beta-catenin-mediated signaling can play an important role in establishing a stem cell hierarchy during crypt morphogenesis. PMID- 11854294 TI - Platelet-derived growth factor (PDGF)-induced tyrosine phosphorylation of the low density lipoprotein receptor-related protein (LRP). Evidence for integrated co receptor function betwenn LRP and the PDGF. AB - The low density lipoprotein receptor-related protein (LRP) functions in the catabolism of numerous ligands including proteinases, proteinase inhibitor complexes, and lipoproteins. In the current study we provide evidence indicating an expanded role for LRP in modulating cellular signaling events. Our results show that platelet-derived growth factor (PDGF) BB induces a transient tyrosine phosphorylation of the LRP cytoplasmic domain in a process dependent on PDGF receptor activation and c-Src family kinase activity. Other growth factors, including basic fibroblast growth factor, epidermal growth factor, insulin-like growth factor-1, were unable to mediate tyrosine phosphorylation of LRP. The basis for this selectivity may result from the ability of LRP to bind PDGFBB, because surface plasmon resonance experiments demonstrated that only PDGF, and not basic fibroblast growth factor, epidermal growth factor, or insulin-like growth factor-1, bound to purified LRP immobilized on a sensor chip. The use of LRP mini-receptor mutants as well as in vitro phosphorylation studies demonstrated that the tyrosine located within the second NPXY motif found in the LRP cytoplasmic domain is the primary site of tyrosine phosphorylation by Src and Src family kinases. Co-immunoprecipitation experiments revealed that PDGF mediated tyrosine phosphorylation of LRPs cytoplasmic domain results in increased association of the adaptor protein Shc with LRP and that Shc recognizes the second NPXY motif within LRPs cytoplasmic domain. In the accompanying paper, Boucher et al. (Boucher, P., Liu, P. V., Gotthardt, M., Hiesberger, T., Anderson, R. G. W., and Herz, J. (2002) J. Biol. Chem. 275, 15507-15513) reveal that LRP is found in caveolae along with the PDGF receptor. Together, these studies suggest that LRP functions as a co-receptor that modulates signal transduction pathways initiated by the PDGF receptor. PMID- 11854295 TI - Platelet-derived growth factor mediates tyrosine phosphorylation of the cytoplasmic domain of the low Density lipoprotein receptor-related protein in caveolae. AB - The low density lipoprotein (LDL) receptor gene family represents a class of multifunctional, endocytic cell surface receptors. Recently, roles in cellular signaling have also emerged. For instance, the very low density lipoprotein receptor (VLDLR) and the apolipoprotein receptor-2 (apoER2) function in a developmental signaling pathway that regulates the lamination of cortical layers in the brain and involves the activation of tyrosine kinases. Furthermore, the cytoplasmic domain of the LDL receptor-related protein (LRP) was found to be a substrate for the non-receptor tyrosine kinase Src, but the physiological significance of this phosphorylation event remained unknown. Here we show that tyrosine phosphorylation of LRP occurs in caveolae and involves the platelet derived growth factor (PDGF) receptor beta and phosphoinositide 3-kinase. Receptor-associated protein, an antagonist of ligand binding to LRP, and apoE enriched beta-VLDL, a ligand for LRP, reduce PDGF-induced tyrosine phosphorylation of the LRP cytoplasmic domain. In the accompanying paper (Loukinova, E., Ranganathan, S., Kuznetsov, S., Gorlatova, N., Migliorini, M., Ulery, P. G., Mikhailenko, I., Lawrence, D. L., and Strickland, D. K. (2002) J. Biol. Chem. 277, 15499-15506) Loukinova et al. further demonstrate that one form of PDGF, PDGF-BB, binds specifically to LRP and that phosphorylation of LRP requires the activation of Src family kinases. Taken together, these findings provide a biochemical basis for a cellular signaling pathway that involves apoE and LRP. PMID- 11854296 TI - Caspase inhibition activates HIV in latently infected cells. Role of tumor necrosis factor receptor 1 and CD95. AB - Stimulation of tumor necrosis factor receptor 1 (TNF-R1) triggers both caspase dependent and caspase-independent signaling activities. The caspase-dependent signaling pathway induces apoptotic cell death in susceptible cells, whereas the caspase-independent signaling cascade leads to activation of nuclear factor kappa B and induces antiapoptotic signaling activities. Stimulation of nuclear factor kappa B via TNF-R1 is known to activate human immunodeficiency virus (HIV) replication in infected cells. Here we show that the broad range caspase inhibitor benzyloxycarbonyl-Val-Ala-Asp-fluoromethyl ketone (ZVAD) activates HIV replication in the chronically infected T-cell line ACH-2. Virus activation was caused by a sensitization of TNF-R1 toward endogenously produced tumor necrosis factor alpha (TNF-alpha). Neutralizing anti-TNF-alpha antibodies completely abolished the virus-inducing activity of ZVAD. Treatment of cells with TNF-alpha in the presence of ZVAD caused increased expression of TNF-alpha and induced enhanced virus replication. Activation of CD95, another member of the TNF receptor family, similarly triggered HIV replication, which was further enhanced in the presence of ZVAD. Our data show that caspase inhibitors sensitize both CD95 and TNF-R1 to mediate activation of HIV in latently infected cells. Activation of HIV replication in latent virus reservoirs is currently discussed as a therapeutic strategy to achieve eradication of HIV in patients treated with antiretroviral therapy. Our results point to a novel role for caspase inhibitors as activators of virus replication in vivo. PMID- 11854297 TI - Signal transductions induced by bone morphogenetic protein-2 and transforming growth factor-beta in normal human osteoblastic cells. AB - Transforming growth factor beta (TGF-beta) activates Ras/MAPK signaling in many cell types. Because TGF-beta and BMP-2 exert similar effects, we examined if this signaling is stimulated by both factors and analyzed the relationship between this signaling and the Smads in osteoblasts. BMP-2 and TGF-beta stimulated Ras, MAPK, and AP-1 activities. The DNA binding activities of c-Fos, FosB/Delta FosB, Fra-1, Fra-2, and JunB were up-regulated whereas JunD activity was decreased. c Fos, FosB/Delta FosB, and JunB were associated with Smad4. The stimulation of AP 1 by BMP-2 and TGF-beta was dependent on Smad signaling, and anti-Smad4 antibody interfered with AP-1 activity. Thus, BMP-2 and TGF-beta activate both Ras/MAPK/AP 1 and Smad signaling in osteoblasts with Smads modulating AP-1 activity. To determine the roles of MAPK in BMP-2 and TGF-beta function, we analyzed the effect of ERK and p38 inhibitors on the regulation of bone matrix protein expression and JunB and JunD levels by these two factors. ERK and p38 mediated TGF-beta suppression of osteocalcin and JunD as well as stimulation of JunB. p38 was essential in BMP-2 up-regulation of type I collagen, fibronectin, osteopontin, osteocalcin, and alkaline phosphatase activity whereas ERK mediated BMP-2 stimulation of fibronectin and osteopontin. Thus, ERK and p38 differentially mediate TGF-beta and BMP-2 function in osteoblasts. PMID- 11854298 TI - The PGC-1-related protein PERC is a selective coactivator of estrogen receptor alpha. AB - Peroxisome proliferator-activated receptor gamma coactivator-1 (PGC-1) is a tissue-specific coactivator that enhances the activity of many nuclear receptors and coordinates transcriptional programs important for energy metabolism. We describe here a novel PGC-1-related coactivator that is expressed in a similar tissue-specific manner as PGC-1, with the highest levels in heart and skeletal muscle. In contrast to PGC-1, the new coactivator shows high receptor specificity. It enhances potently the activity of estrogen receptor (ER) alpha, while having only small effects on other receptors. Because of its nuclear receptor selectivity, we have termed the new protein PERC (PGC-1 related Estrogen Receptor Coactivator). We show here that the coactivation function of PERC relies on a bipartite transcriptional activation domain and two LXXLL motifs that interact with the AF2 domain of ERalpha in an estrogen-dependent manner. PERC and PGC-1 are likely to have different functions in ER signaling. Whereas PERC acts selectively on ERalpha and not on the second estrogen receptor ERbeta, PGC-1 coactivates strongly both ERs. Moreover, PERC and PGC-1 show distinct preferences for enhancing ERalpha in different promoter contexts. Finally, PERC enhances the ERalpha-mediated response to the partial agonist tamoxifen, while PGC-1 modestly represses it. The two coactivators are likely to mediate distinct, tissue specific responses to estrogens. PMID- 11854299 TI - Impaired receptor binding and activation associated with a human prostacyclin receptor polymorphism. AB - The human prostacyclin receptor (hIP) is a seven transmembrane-spanning G-protein coupled receptor that plays an important role in vascular homeostasis. Recent genetic analyses (SNP database, NCBI) have revealed the first two polymorphisms within the coding sequence, V25M and R212H. Here we present structure-function characterizations of these polymorphisms at physiological pH (7.4) and at an acidic pH (6.8) that would be encountered during stress such as renal, respiratory, or heart failure. Through a series of competition binding and G protein activation assays (measured by cAMP production), we determined that the V25M polymorph exhibited agonist binding and G-protein activation similar to wild type receptor at normal pH (7.4). However, the R212H variant demonstrated a significant decrease in binding affinity at lower pH (R212H at pH 7.4, K(i) = 2.2 +/- 1.2 nm; pH 6.8 K(i) = 45.6 +/- 12.0 nm). The R212H polymorph also exhibited abnormal activation at both pH 7.4 and pH 6.8 (pH 7.4, R212H EC(50) = 2.8 +/- 0.5 nm versus wild-type hIP EC(50) = 0.5 +/- 0.1 nm; pH 6.8, R212H EC(50) = 3.2 +/- 1.6 nm versus wild-type hIP EC(50) = 0.5 +/- 0.2 nm). Polymorphisms of the human prostacyclin receptor potentially may be important predictors of disease progress during biological stressors such as acidosis in which urgent correction of bodily pH may be required to restore normal hemostasis and vasodilation. This study provides the mechanistic basis for further research into genetic risk factors and pharmacogenetics of cardiovascular disease associated with hIP. PMID- 11854300 TI - Mechanism of human telomerase inhibition by BIBR1532, a synthetic, non nucleosidic drug candidate. AB - Telomerase, a ribonucleoprotein acting as a reverse transcriptase, has been identified as a target for cancer drug discovery. The synthetic, non-nucleosidic compound, BIBR1532, is a potent and selective telomerase inhibitor capable of inducing senescence in human cancer cells (). In the present study, the mode of drug action was characterized. BIBR1532 inhibits the native and recombinant human telomerase, comprising the human telomerase reverse transcriptase and human telomerase RNA components, with similar potency primarily by interfering with the processivity of the enzyme. Enzyme-kinetic experiments show that BIBR1532 is a mixed-type non-competitive inhibitor and suggest a drug binding site distinct from the sites for deoxyribonucleotides and the DNA primer, respectively. Thus, BIBR1532 defines a novel class of telomerase inhibitor with mechanistic similarities to non-nucleosidic inhibitors of HIV1 reverse transcriptase. PMID- 11854301 TI - Mapping of the dimer interface of the Escherichia coli mannitol permease by cysteine cross-linking. AB - A cysteine cross-linking approach was used to identify residues at the dimer interface of the Escherichia coli mannitol permease. This transport protein comprises two cytoplasmic domains and one membrane-embedded C domain per monomer, of which the latter provides the dimer contacts. A series of single-cysteine His tagged C domains present in the native membrane were subjected to Cu(II)-(1,10 phenanthroline)(3)-catalyzed disulfide formation or cysteine cross-linking with dimaleimides of different length. The engineered cysteines were at the borders of the predicted membrane-spanning alpha-helices. Two residues were found to be located in close proximity of each other and capable of forming a disulfide, while four other locations formed cross-links with the longer dimaleimides. Solubilization of the membranes did only influence the cross-linking behavior at one position (Cys(73)). Mannitol binding only effected the cross-linking of a cysteine at the border of the third transmembrane helix (Cys(134)), indicating that substrate binding does not lead to large rearrangements in the helix packing or to dissociation of the dimer. Upon mannitol binding, the Cys(134) becomes more exposed but the residue is no longer capable of forming a stable disulfide in the dimeric IIC domain. In combination with the recently obtained projection structure of the IIC domain in two-dimensional crystals, a first proposal is made for alpha-helix packing in the mannitol permease. PMID- 11854302 TI - Oligomerization of G-protein-coupled receptors shown by selective co immunoprecipitation. AB - Recent studies have shown that G-protein-coupled receptors (GPCRs) can assemble as high molecular weight homo- and hetero-oligomeric complexes. This can result in altered receptor-ligand binding, signaling, or intracellular trafficking. We have co-transfected HEK-293 cells with differentially epitope-tagged GPCRs from different subfamilies and determined whether oligomeric complexes were formed by co-immunoprecipitation and immunoblot analysis. This gave the surprising result that the 5HT(1A) receptor was capable of forming hetero-oligomers with all GPCRs tested including the 5HT(1B), 5HT(1D), EDG(1), EDG(3), GPR(26), and GABA(B2) receptors. The testing of other GPCR combinations showed similar results with hetero-oligomer formation occurring for the 5HT(1D) with the 5HT(1B) and EDG(1) receptor. Control studies showed that these complexes were present in co transfected cells before the time of lysis and that the hetero-oligomers were comprised of GPCRs at discrete stoichiometries. These findings suggest that GPCRs have a natural tendency to form oligomers when co-transfected into cells. Future studies should therefore investigate the presence and physiological role of GPCR hetero-oligomers in cells in which they are endogenously expressed. PMID- 11854303 TI - Mechanism of interaction between leucine-based sorting signals from the invariant chain and clathrin-associated adaptor protein complexes AP1 and AP2. AB - The cytoplasmic tail of the invariant chain contains two leucine-based sorting signals, and each of those seems sufficient to route the invariant chain to its intracellular destination in either normal or polarized cells. It is believed that the intracellular routing of the invariant chain is mediated by its interactions with the clathrin-associated adaptor protein complexes AP1 and AP2. We () have previously demonstrated the in vitro interactions between the cytoplasmic tail of the invariant chain and AP1/AP2 complexes. These interactions were specific and depended on the critical leucine residues in the invariant chain's sorting signals. In the present study, we decided to investigate the molecular mechanism of these interactions. To this end, we constructed a set of glutathione S-transferase fusion proteins that contained the intact cytoplasmic tail of the invariant chain and its various mutants to define residues important for its interactions with AP1 and AP-2. Our results demonstrated the importance of several residues other than the critical leucine residues for such interactions. A strong correlation between in vitro binding of AP2 to the invariant chain and in vivo internalization of the invariant chain was observed, confirming the primary role of AP2 in recognition of endocytic signals. In addition, we demonstrated different requirements for AP1 and AP2 binding to cytoplasmic tail of the invariant chain, which may reflect that the different sorting pathways mediated by AP1 and AP2 involve their recognition of the primary structure of the sorting signal. PMID- 11854304 TI - Extending the Arp2/3 complex and its regulation beyond the leading edge. AB - Two studies characterizing Drosophila Arp2/3 complex and Scar mutants demonstrate that assembly of some actin structures in nonmotile cells of multicellular organisms utilizes the same proteins as are important for actin assembly in motile cells. These studies also show that assembly of other actin structures is independent of these proteins, suggesting that alternative mechanisms also exist. PMID- 11854305 TI - SUMO-1 targets RanGAP1 to kinetochores and mitotic spindles. AB - RanGAP1 was the first documented substrate for conjugation with the ubiquitin like protein SUMO-1. However, the functional significance of this conjugation has not been fully clarified. We sought to examine RanGAP1 behavior during mitosis. We found that RanGAP1 associates with mitotic spindles and that it is particularly concentrated at foci near kinetochores. Association with kinetochores appeared soon after nuclear envelope breakdown and persisted until late anaphase, but it was lost coincident with nuclear envelope assembly in telophase. A mutant RanGAP1 protein lacking the capacity to be conjugated to SUMO 1 no longer associated with spindles, indicating that conjugation was essential for RanGAP1's mitotic localization. RanBP2, a nuclear pore protein that binds SUMO-1-conjugated RanGAP1 during interphase, colocalized with RanGAP1 on spindles, suggesting that a complex between these two proteins may be involved in mitotic targeting of RanGAP1. This report shows for the first time that SUMO-1 conjugation is required for mitotic localization of RanGAP1, and suggests that a major role of SUMO-1 conjugation to RanGAP1 may be the spatial regulation of the Ran pathway during mitosis. PMID- 11854306 TI - Alteration of nuclear lamin organization inhibits RNA polymerase II-dependent transcription. AB - Regulation of gene activity is mediated by alterations in chromatin organization. In addition, chromatin organization may be governed in part by interactions with structural components of the nucleus. The nuclear lamins comprise the lamina and a variety of nucleoplasmic assemblies that together are major structural components of the nucleus. Furthermore, lamins and lamin-associated proteins have been reported to bind chromatin. These observations suggest that the nuclear lamins may be involved in the regulation of gene activity. In this report, we test this possibility by disrupting the normal organization of nuclear lamins with a dominant negative lamin mutant lacking the NH2-terminal domain. We find that this disruption inhibits RNA polymerase II activity in both mammalian cells and transcriptionally active embryonic nuclei from Xenopus laevis. The inhibition appears to be specific for polymerase II as disruption of lamin organization does not detectably inhibit RNA polymerases I and III. Furthermore, immunofluorescence observations indicate that this selective inhibition of polymerase II-dependent transcription involves the TATA binding protein, a component of the basal transcription factor TFIID. PMID- 11854307 TI - CLIPR-59, a new trans-Golgi/TGN cytoplasmic linker protein belonging to the CLIP 170 family. AB - The microtubule cytoskeleton plays a fundamental role in cell organization and membrane traffic in higher eukaryotes. It is well established that molecular motors are involved in membrane-microtubule interactions, but it has also been proposed that nonmotor microtubule-binding (MTB) proteins known as CLIPs (cytoplasmic linker proteins) have basic roles in these processes. We report here the characterization of CLIPR-59, a CLIP-170-related protein localized to the trans-most part of the Golgi apparatus. CLIPR-59 contains an acidic region followed by three ankyrin-like repeats and two CLIP-170-related MTB motifs. We show that the 60-amino acid-long carboxy-terminal domain of CLIPR-59 is necessary and sufficient to achieve Golgi targeting, which represents the first identification of a membrane targeting domain in a CLIP-170-related protein. The MTB domain of CLIPR-59 is functional because it localizes to microtubules when expressed as a fragment in HeLa cells. However, our results suggest that this domain is normally inhibited by the presence of adjacent domains, because neither full-length CLIPR-59 nor a CLIPR-59 mutant missing its membrane-targeting region localize to microtubules. Consistent with this observation, overexpression of CLIPR-59 does not affect the microtubule network. However, CLIPR-59 overexpression strongly perturbs early/recycling endosome-TGN dynamics, implicating CLIPR-59 in the regulation of this pathway. PMID- 11854308 TI - A subset of dynamic actin rearrangements in Drosophila requires the Arp2/3 complex. AB - The Arp2/3 complex has been shown to dramatically increase the slow spontaneous rate of actin filament nucleation in vitro, and it is known to be important for remodeling the actin cytoskeleton in vivo. We isolated and characterized loss of function mutations in genes encoding two subunits of the Drosophila Arp2/3 complex: Arpc1, which encodes the homologue of the p40 subunit, and Arp3, encoding one of the two actin-related proteins. We used these mutations to study how the Arp2/3 complex contributes to well-characterized actin structures in the ovary and the pupal epithelium. We found that the Arp2/3 complex is required for ring canal expansion during oogenesis but not for the formation of parallel actin bundles in nurse cell cytoplasm and bristle shaft cells. The requirement for Arp2/3 in ring canals indicates that the polymerization of actin filaments at the ring canal plasma membrane is important for driving ring canal growth. PMID- 11854309 TI - SCAR is a primary regulator of Arp2/3-dependent morphological events in Drosophila. AB - The Arp2/3 complex and its activators, Scar/WAVE and Wiskott-Aldrich Syndrome protein (WASp), promote actin polymerization in vitro and have been proposed to influence cell shape and motility in vivo. We demonstrate that the Drosophila Scar homologue, SCAR, localizes to actin-rich structures and is required for normal cell morphology in multiple cell types throughout development. In particular, SCAR function is essential for cytoplasmic organization in the blastoderm, axon development in the central nervous system, egg chamber structure during oogenesis, and adult eye morphology. Highly similar developmental requirements are found for subunits of the Arp2/3 complex. In the blastoderm, SCAR and Arp2/3 mutations result in a reduction in the amount of cortical filamentous actin and the disruption of dynamically regulated actin structures. Remarkably, the single Drosophila WASp homologue, Wasp, is largely dispensable for these numerous Arp2/3-dependent functions, whereas SCAR does not contribute to cell fate decisions in which Wasp and Arp2/3 play an essential role. These results identify SCAR as a major component of Arp2/3-dependent cell morphology during Drosophila development and demonstrate that the Arp2/3 complex can govern distinct cell biological events in response to SCAR and Wasp regulation. PMID- 11854310 TI - Drosophila Kelch regulates actin organization via Src64-dependent tyrosine phosphorylation. AB - The Drosophila kelch gene encodes a member of a protein superfamily defined by the presence of kelch repeats. In Drosophila, Kelch is required to maintain actin organization in ovarian ring canals. We set out to study the actin cross-linking activity of Kelch and how Kelch function is regulated. Biochemical studies using purified, recombinant Kelch protein showed that full-length Kelch bundles actin filaments, and kelch repeat 5 contains the actin binding site. Two-dimensional electrophoresis demonstrated that Kelch is tyrosine phosphorylated in a src64 dependent pathway. Site-directed mutagenesis determined that tyrosine residue 627 is phosphorylated. A Kelch mutant with tyrosine 627 changed to alanine (KelY627A) rescued the actin disorganization phenotype of kelch mutant ring canals, but failed to produce wild-type ring canals. Electron microscopy demonstrated that phosphorylation of Kelch is critical for the proper morphogenesis of actin during ring canal growth, and presence of the nonphosphorylatable KelY627A protein phenocopied src64 ring canals. KelY627A protein in ring canals also dramatically reduced the rate of actin monomer exchange. The phenotypes caused by src64 mutants and KelY627A expression suggest that a major function of Src64 signaling in the ring canal is the negative regulation of actin cross-linking by Kelch. PMID- 11854311 TI - Coordination of opposite-polarity microtubule motors. AB - Many cargoes move bidirectionally, frequently reversing course between plus- and minus-end microtubule travel. For such cargoes, the extent and importance of interactions between the opposite-polarity motors is unknown. In this paper we test whether opposite-polarity motors on lipid droplets in Drosophila embryos are coordinated and avoid interfering with each other's activity, or whether they engage in a tug of war. To this end we impaired the minus-end transport machinery using dynein and dynactin mutations, and then investigated whether plus-end motion was improved or disrupted. We observe a surprisingly severe impairment of plus-end motion due to these alterations of minus-end motor activity. These observations are consistent with a coordination hypothesis, but cannot be easily explained with a tug of war model. Our measurements indicate that dynactin plays a crucial role in the coordination of plus- and minus-end-directed motors. Specifically, we propose that dynactin enables dynein to participate efficiently in bidirectional transport, increasing its ability to stay "on" during minus-end motion and keeping it "off" during plus-end motion. PMID- 11854312 TI - T helper subset development: roles of instruction, selection, and transcription. PMID- 11854313 TI - Death receptors couple to both cell proliferation and apoptosis. PMID- 11854314 TI - Proteotoxicity in the endoplasmic reticulum: lessons from the Akita diabetic mouse. PMID- 11854315 TI - Genetic models: clues for understanding the pathogenesis of idiopathic nephrotic syndrome. PMID- 11854316 TI - Transcription factor haploinsufficiency: when half a loaf is not enough. PMID- 11854317 TI - Leydig cell-derived heme oxygenase-1 regulates apoptosis of premeiotic germ cells in response to stress. AB - Stress-induced downregulation of spermatogenesis remains poorly understood. This study examined the induction of heme oxygenase-1 (HO-1), a carbon monoxide generating inducible enzyme, in modulation of spermatogenesis. Rats were exposed to cadmium chloride (CdCl(2)), a stressor causing oligozoospermia, and HO-1 induction was monitored by following HO isozyme expression. CdCl(2)-treated testes increased HO-1 activity and suppressed microsomal cytochromes P450, which are required for steroidogenesis. CdCl(2)-elicited HO-1 occurred mostly in Leydig cells and coincided with CO generation, as judged by bilirubin-IXalpha immunoreactivity. Under these circumstances, germ cells in peripheral regions of seminiferous tubules exhibited apoptosis; laser flow cytometry revealed that these apoptotic cells involve diploid and tetraploid germ cells, suggesting involvement of spermatogonia and primary spermatocytes in CdCl(2)-elicited apoptosis. Pretreatment with zinc protoporphyrin-IX, an HO inhibitor, but not copper protoporphyrin-IX, which does not block the enzyme, attenuated the CdCl(2) induced apoptosis. Such antiapoptotic effects of zinc protoporphyrin-IX were repressed by supplementation of dichloromethane, a CO donor. Upon CdCl(2) treatment, both Sertoli cells and the germ cells upregulated Fas ligand; this event was also suppressed by zinc protoporphyrin-IX and restored by dichloromethane. Thus, Leydig cells appear to use HO-1-derived CO to trigger apoptosis of premeiotic germ cells and thereby modulate spermatogenesis under conditions of stress. PMID- 11854318 TI - Partial deficiency of thyroid transcription factor 1 produces predominantly neurological defects in humans and mice. AB - Three genes, TTF1, TTF2, and PAX8, involved in thyroid gland development and migration have been identified. Yet systematic screening for defects in these genes in thyroid dysgenesis gave essentially negative results. In particular, no TTF1 gene defects were found in 76 individuals with thyroid dysgenesis even though a deletion of this gene in the mouse results in thyroid and lung agenesis and defective diencephalon. We report a 6-year-old boy with predominant dyskinesia, neonatal respiratory distress, and mild hyperthyrotropinemia. One allele of his TTF1 gene had a guanidine inserted into codon 86 producing a nonsense protein of 407, rather than 371, amino acids. The mutant TTF1 did not bind to its canonical cis-element or transactivate a reporter gene driven by the thyroglobulin promoter, a natural target of TTF1. Failure of the mutant TTF1 to interfere with binding and transactivation functions of the wild-type TTF1 suggested that the syndrome was caused by haploinsufficiency. This was confirmed in mice heterozygous for Ttf1 gene deletion, heretofore considered to be normal. Compared with wild-type littermates, Ttf1(+/-) mice had poor coordination and a significant elevation of serum thyrotropin. Therefore, haploinsufficiency of the TTF1 gene results in a predominantly neurological phenotype and secondary hyperthyrotropinemia. PMID- 11854319 TI - Choreoathetosis, hypothyroidism, and pulmonary alterations due to human NKX2-1 haploinsufficiency. AB - The occurrence of neurological symptoms and developmental delay in patients affected by congenital hypothyroidism (CH) has been attributed to the lack of thyroid hormone in the developing CNS. Accordingly, after the introduction of neonatal screening programs for CH, which allowed early and adequate treatment, an almost normal outcome for most CH patients could be achieved. However, a few patients did not reach this favorable outcome despite early and adequate treatment. Here we describe five patients with variable degrees of CH who suffered from choreoathetosis, muscular hypotonia, and pulmonary problems, an association of symptoms that had not been described before this study. Since this clinical picture matched the phenotype of mice targeted for deletion of the transcription factor gene Nkx2-1, we investigated the human NKX2-1 gene in these five patients. We found heterozygous loss of function mutations in each of these five patients, e.g., one complete gene deletion, one missense mutation (G2626T), and three nonsense mutations (2595insGG, C2519A, C1302A). Therefore, the unfavorable outcome in patients with CH, especially those with choreoathetosis and pulmonary symptoms, can be explained by mutations in the NKX2-1 gene rather than by hypothyroidism. Moreover, the association of symptoms in the patients with NKX2-1 mutations points to an important role of human NKX2-1 in the development and function of thyroid, basal ganglia, and lung, as already described for rodents. PMID- 11854320 TI - The polycystin-1 C-terminal fragment triggers branching morphogenesis and migration of tubular kidney epithelial cells. AB - Mutations of either PKD1 or PKD2 cause autosomal dominant polycystic kidney disease, a syndrome characterized by extensive formation of renal cysts and progressive renal failure. Homozygous deletion of Pkd1 or Pkd2, the genes encoding polycystin-1 and polycystin-2, disrupt normal renal tubular differentiation in mice but do not affect the early steps of renal development. Here, we show that expression of the C-terminal 112 amino acids of human polycystin-1 triggers branching morphogenesis and migration of inner medullary collecting duct (IMCD) cells, and support in vitro tubule formation. The integrity of the polycystin-2-binding region is necessary but not sufficient to induce branching of IMCD cells. The C-terminal domain of polycystin-1 stimulated protein kinase C-alpha (PKC-alpha), but not the extracellular signal-regulated kinases ERK1 or ERK2. Accordingly, inhibition of PKC, but not ERK, prevented polycystin-1-mediated IMCD cell morphogenesis. In contrast, HGF-mediated morphogenesis required ERK activation but was not dependent on PKC. Our findings demonstrate that the C-terminal domain of polycystin-1, acting in a ligand independent fashion, triggers unique signaling pathways for morphogenesis, and likely plays a central role in polycystin-1 function. PMID- 11854321 TI - Extrarenal effects on the pathogenesis and relapse of idiopathic nephrotic syndrome in Buffalo/Mna rats. AB - Buffalo/Mna rats spontaneously develop a focal segmental glomerulosclerosis with a histological pattern similar to the human disease. In this study, we investigated the potential of recurrence of the disease by transplantation of normal kidneys into Buffalo/Mna recipients. Kidneys from healthy LEW.1W rats were grafted into proteinuric 6-month-old Buffalo/Mna rats without or with specific tolerance induction following donor-specific transfusion (DST) aimed at controlling host anti-donor immune responses. The inverse combination was carried out to determine whether a proteinuric Buffalo/Mna kidney can recover its permselectivity in a normal environment. As a control, LEW.1W kidneys were grafted into Wistar Furth recipients. After transplantation without DST, recurrence of proteinuria in LEW.1W kidneys appeared at approximately 10 days, possibly associated with rejection of the graft. In the same combination with DST, proteinuria occurred after 20 days, and the attendant glomerular damage suggested that the initial kidney disease had recurred. Transplanted control animals remained free of proteinuria. In the opposite combination, the proteinuria and the lesions of Buffalo/Mna kidneys regressed after transplantation into healthy LEW.1W rats. The recurrence of proteinuria after transplantation in Buffalo/Mna and the remission of lesions in Buffalo/Mna kidneys transplanted into normal hosts suggests that Buffalo/Mna rats express circulating albuminuric factors, which may be relevant to the relapse of idiopathic nephrotic syndrome in humans. PMID- 11854322 TI - Formation of protein kinase C(epsilon)-Lck signaling modules confers cardioprotection. AB - The epsilon isoform of protein kinase C (PKCepsilon) is a member of the PKC family of serine/threonine kinases and plays a critical role in protection against ischemic injury in multiple organs. Functional proteomic analyses of PKCepsilon signaling show that this isozyme forms multiprotein complexes in the heart; however, the precise signaling mechanisms whereby PKCepsilon orchestrates cardioprotection are poorly understood. Here we report that Lck, a member of the Src family of tyrosine kinases, forms a functional signaling module with PKCepsilon. In cardiac cells, PKCepsilon interacts with, phosphorylates, and activates Lck. In vivo studies showed that cardioprotection elicited either by cardiac-specific transgenic activation of PKCepsilon or by ischemic preconditioning enhances the formation of PKCepsilon-Lck modules. Disruption of these modules, via ablation of the Lck gene, abrogated the infarct-sparing effects of these two forms of cardioprotection, indicating that the formation of PKCepsilon-Lck signaling modules is required for the manifestation of a cardioprotective phenotype. These findings demonstrate, for the first time to our knowledge, that the assembly of a module (PKCepsilon-Lck) is an obligatory step in the signal transduction that results in a specific phenotype. Thus, PKCepsilon Lck modules may serve as novel therapeutic targets for the prevention of ischemic injury. PMID- 11854323 TI - Role of sarcolemmal K(ATP) channels in cardioprotection against ischemia/reperfusion injury in mice. AB - Recently it has been postulated that mitochondrial ATP-sensitive K(+) (mitoK(ATP)) channels rather than sarcolemmal K(ATP) (sarcK(ATP)) channels are important as end effectors and/or triggers of ischemic preconditioning (IPC). To define the pathophysiological significance of sarcK(ATP) channels, we conducted functional experiments using Kir6.2-deficient (KO) mice. Metabolic inhibition with glucose-free, dinitrophenol-containing solution activated sarcK(ATP) current and shortened the action potential duration in ventricular cells isolated from wild-type (WT) but not KO mice. MitoK(ATP) channel function was preserved in KO ventricular cells. In anesthetized mice, IPC reduced the infarct size in WT but not KO mice. Following global ischemia/reperfusion, the increase of left ventricular end-diastolic pressure during ischemia was more marked, and the recovery of contractile function was worse, in KO hearts than in WT hearts. Treatment with HMR1098, a sarcK(ATP) channel blocker, but not 5-hydroxydecanoate, a mitoK(ATP) channel blocker, produced a deterioration of contractile function in WT hearts comparable to that of KO hearts. These findings suggest that sarcKATP channels figures prominently in modulating ischemia/reperfusion injury in the mouse. The rapid heart rate of the mouse (>600 beats per minute) may magnify the relative importance of sarcK(ATP) channels during ischemia, prompting caution in the extrapolation of the conclusions to larger mammals. PMID- 11854324 TI - CD4(+) T cells specific to a glomerular basement membrane antigen mediate glomerulonephritis. AB - Ab-mediated mechanisms have been considered the major causes of glomerulonephritis (GN). However, recent studies suggest that T cells may be more important in mediating GN. To investigate the effects of antigen-specific CD4(+) T cells, we generated Th1 cell lines specific for this antigen from rats that had been immunized with a recombinant form of the glomerular basement membrane (GBM) antigen, Col4alpha3NC1. Upon the transfer of in vitro-activated T cell lines to pertussis toxin-primed, naive syngeneic rats, the recipients developed severe proteinuria/albuminuria, which plateaued after approximately 35 days. Although no IgG binding to GBM or C3 deposition could be detected by immunofluorescence, five out of eleven rats exhibited severe GN, as judged by the formation of characteristic crescent-shaped lesions in the glomeruli, whereas the others exhibited modest GN. Thus Col4alpha3NC1-specific T cells directly initiated glomerular injury in the recipients. One notable difference from GN induced by active immunization was a T cell infiltration in the renal interstitium, which affected some tubules. We therefore injected fluorescence-labeled Col4alpha3NC1 specific into naive rats, and we found that they were enriched 4.5-fold in the kidney cortex relative to nonspecific control T cells 24 hours later. Many of the T cells were located in the Bowman's space and had a flattened shape, suggesting that the primary target for the T cells was in or adjacent to the Bowman's capsule. PMID- 11854325 TI - Targeted disruption of the Chop gene delays endoplasmic reticulum stress-mediated diabetes. AB - Overload of pancreatic beta cells in conditions such as hyperglycemia, obesity, and long-term treatment with sulfonylureas leads to beta cell exhaustion and type 2 diabetes. Because beta cell mass declines under these conditions, apparently as a result of apoptosis, we speculated that overload kills beta cells as a result of endoplasmic reticulum (ER) stress. The Akita mouse, which carries a conformation-altering missense mutation (Cys96Tyr) in Insulin 2, likewise exhibits hyperglycemia and a reduced beta cell mass. In the development of diabetes in Akita mice, mRNAs for the ER chaperone Bip and the ER stress associated apoptosis factor Chop were induced in the pancreas. Overexpression of the mutant insulin in mouse MIN6 beta cells induced Chop expression and led to apoptosis. Targeted disruption of the Chop gene delayed the onset of diabetes in heterozygous Akita mice by 8-10 weeks. We conclude that ER overload in beta cells causes ER stress and leads to apoptosis via Chop induction. Our findings suggest a new therapeutic approach for preventing the onset of diabetes by inhibiting Chop induction or by increasing chaperone capacity in the ER. PMID- 11854327 TI - Interactions between endothelial nitric oxide synthase and sex hormones in vascular protection in mice. AB - The vasculoprotective effects of sex hormones, particularly estrogens, have been attributed to their ability to increase the bioavailability of nitric oxide through activation of endothelial nitric oxide synthase (eNOS). To dissect the relative contribution in vivo of eNOS, sex hormones, and their interaction in two complex vascular phenotypes, hypertension and atherosclerosis, we used mice doubly deficient in eNOS and apoE (nnee) or lacking only apoE (NNee). Females and males were gonadectomized at 1 month of age and implanted either with control pellets or pellets releasing 17beta-estradiol (E2). Hormonally intact nnee mice have elevated blood pressure (BP) and increased atherosclerosis compared with NNee mice, but on removal of gonads, BP and atherosclerosis decreased significantly in nnee mice but not in NNee mice. Three months of treatment with exogenous E2 dramatically reduced atherosclerosis and significantly lowered BP in both NNee and nnee mice compared with animals treated with control pellets. Thus exogenous E2 has strong BP-lowering and atheroprotective effects in apoE deficient mice, but eNOS is not essential for either effect. Endogenous sex hormones, on the other hand, cause significant damage to the vasculature in the absence of eNOS, but these effects are overridden by interactions between eNOS and sex hormones. PMID- 11854326 TI - Cystin, a novel cilia-associated protein, is disrupted in the cpk mouse model of polycystic kidney disease. AB - The congenital polycystic kidney (cpk) mutation is the most extensively characterized mouse model of polycystic kidney disease (PKD). The renal cystic disease is fully expressed in homozygotes and is strikingly similar to human autosomal recessive PKD (ARPKD), whereas genetic background modulates the penetrance of the corresponding defect in the developing biliary tree. We now describe the positional cloning, mutation analysis, and expression of a novel gene that is disrupted in cpk mice. The cpk gene is expressed primarily in the kidney and liver and encodes a hydrophilic, 145-amino acid protein, which we term cystin. When expressed exogenously in polarized renal epithelial cells, cystin is detected in cilia, and its expression overlaps with polaris, another PKD-related protein. We therefore propose that the single epithelial cilium is important in the functional differentiation of polarized epithelia and that ciliary dysfunction underlies the PKD phenotype in cpk mice. PMID- 11854331 TI - From North Sea to Black Sea: progress towards common public health practice. PMID- 11854332 TI - Validation of self reported smoking. PMID- 11854333 TI - Income inequality and economic residential segregation. PMID- 11854328 TI - Blockade of LIGHT/LTbeta and CD40 signaling induces allospecific T cell anergy, preventing graft-versus-host disease. AB - Previous studies have shown that blockade of LIGHT, a T cell costimulatory molecule belonging to the TNF superfamily, by soluble lymphotoxin beta receptor Ig (LTbetaR-Ig) inhibits the cytotoxic T lymphocyte (CTL) response to host antigenic disparities and ameliorates lethal graft-versus-host disease (GVHD) in a B6 to BDF1 mouse model. Here, we demonstrate that infusion of an mAb against CD40 ligand (CD40L) further increases the efficacy of LTbetaR-Ig, leading to complete prevention of GVHD. We further demonstrate that alloantigen-specific CTLs become anergic upon rapid expansion, and persist in the tolerized mice as a result of costimulatory blockade. Transfer of anergic CTLs to secondary F1 mice fails to induce GVHD despite the fact that anergic CTLs can be stimulated to proliferate in vitro by antigens and cytokines. Our study provides a potential new approach for the prevention of lethal GVHD. PMID- 11854334 TI - Validation of self reported smoking by serum cotinine measurement in a community based study. AB - STUDY OBJECTIVE: The validity of self reported smoking in population surveys remains an important question. An associated question is what would be the value of measuring serum cotinine concentrations in such surveys to obtain validated smoking data. DESIGN: Cross sectional analysis of data on self reported smoking and serum cotinine among a random population sample of 5846 persons aged 25 to 64 years, who participated in the FINRISK-92 survey. MAIN RESULTS: Among self reported regular smokers, 97.2% of men and 94.9% of women had a cotinine concentration of 10 ng/ml or higher in serum. Of those participants who reported to have smoked at any time during their life but not during the previous month, 6.3% of men and 5.2% of women had a serum cotinine concentration of at least 10 ng/ml. Among never smokers 2.5% of men and 2.7% of women had detectable level of cotinine in their serum. The validity of self reporting was similar among subjects from different areas, ages, and socioeconomic groups. CONCLUSIONS: In a sample of the general population in Finland the validity of self reported smoking is high, and most of the few self reported non-smokers who had cotinine in their serum had only low or moderate levels. PMID- 11854336 TI - Collecting information on marital status: a methodological note. PMID- 11854335 TI - Is the link between alcohol and cardiovascular death among young Russian men attributable to misclassification of acute alcohol intoxication? Evidence from the city of Izhevsk. AB - BACKGROUND: Research on the aetiology of sudden cardiac death among young men in Russia strongly suggests an association with binge drinking. However, the possibility remains that such deaths are misclassified as being attributable to cardiovascular disease when they are really caused by acute alcohol poisoning. OBJECTIVE: To describe postmortem levels of blood alcohol in Russian men dying from various causes and so determine whether deaths from alcohol poisoning are being misclassified as cardiovascular deaths. SETTING: Ishevsk, capital of the Udmurt Republic, situated in the Ural region of the Russian Federation. METHODS: The study was part of a larger one on adult mortality. The study sample was 309 deaths among men aged 20-55 dying between August 1998 and March 1999 from other than neoplasms, infectious diseases or unspecified causes and on whom necropsy records could be obtained. Information on cause of death was extracted from death certificates and data on postmortem blood alcohol concentration (BAC) from forensic records. Blood alcohol concentrations were adjusted where necessary to allow for delay in necropsy. RESULTS: Medium or greater levels of intoxication occurred in a quarter of those recorded as dying from cardiovascular disease but in over half of those dying from external causes. BAC levels consistent with at least strong intoxication were seen in 13.5% of deaths from cardiovascular disease and 27.1% from external causes. No cardiovascular deaths had BAC at levels usually thought to be fatal while this level was seen in 26% of deaths from accidental poisoning. CONCLUSION: Evidence of recent consumption of alcohol is common among Russian men dying under the age of 55, with severe intoxication common where death is from external causes. However, the high death rates from cardiovascular disease in Russia cannot be explained by misclassification of deaths attributable to acute alcohol poisoning. This study thus resolves one of the outstanding controversies in the story of alcohol and cardiovascular disease in the former Soviet Union. PMID- 11854337 TI - On the World Health Organisation's measurement of health inequalities. AB - STUDY OBJECTIVE: To review the World Health Organisation's methodological approach for the purpose of measuring health inequalities presented in the WHR 2000 and reference papers. MAIN FINDINGS: Recommending that health inequalities be assessed by measuring interindividual differences, without regard for the distribution of health status among specific population subgroups, the approach taken by WHO does not take into account the socioeconomic dimension, is strongly influenced by the extent of socioeconomic inequalities in the population, and suffers from the health redistribution problem. Apart from the conceptual issues, the estimation procedure also has methodological problems hidden in a sophisticated statistical procedure, which is confusingly explained in one of the referred discussion papers. The results presented in the WHR 2000 are based on Demographic and Health Survey data that refer to more than 10 years ago. OTHER METHODOLOGICAL PROBLEMS: The WHO's individual differences measure of health inequalities is expressed in units of survival time raised to the power of 2.5. Besides the difficulty of interpretation, the individual differences index is not a relative measure. However, the index of equality of child survival was defined as the complement of the individual differences index, as though it were a relative measure. NEGLECT TO THE SPECIALISED LITERATURE: The WHO's index is a particular case in a family of measures that provides generalisations of the Gini coefficient. However, concerns on the adequacy and validity of this procedure for the purposes of measuring health inequalities were completely ignored. CONCLUSIONS: The need to open up the debate with the scientific community has been recently recognised by the executive board of the WHO. In view of the new prospect, the paper concludes by raising some points that can contribute to the discussion on the measurement of health inequalities, with regard to the evaluation of the health system performance. PMID- 11854338 TI - Inequality, residential segregation by income, and mortality in US cities. AB - STUDY OBJECTIVE: This study was designed to discover whether the relation between income inequality and population death rates within the United States was mediated by the degree of residential segregation between rich and poor. DESIGN: Using data for 276 metropolitan areas in the USA, residential segregation was defined as the extent to which people with different levels of income live in the same or different census tracts. Two segregation measures were used: the ratio of income inequality between household within tracts to the inequality in average income between tracts, and the Jargowsky Neighbourhood Sorting Index. MAIN RESULTS: Results suggest that segregation within urban areas is associated with an additional mortality burden. However, the association between income inequality and mortality in these metropolitan statistical areas was found to be independent of the degree of economic segregation between their constituent neighbourhoods. CONCLUSIONS: Most of the association between income inequality and mortality is not mediated by the effects of greater residential segregation. PMID- 11854339 TI - Class differences in the social consequences of illness? AB - STUDY OBJECTIVE: To investigate adverse social consequences of limiting longstanding illness and the modifying effect of socioeconomic position on these consequences. DESIGN: Cohort study on the panel within the annual Swedish Survey of Living Conditions where participants were interviewed twice with eight years interval 1979-89 and 1986-97. Sociodemographic characteristics, self reported longstanding illness, employment situation and financial conditions were measured at baseline. Social consequences (economic inactivity, unemployment, financial difficulties) of limiting longstanding illness were measured at follow up eight years later. SETTING: National sample for Sweden during a period that partly was characterised by high unemployment and reduction in insurance benefits. PARTICIPANTS: PARTICIPANTS were 13 855 men and women, economically active, not unemployed, without financial difficulties at the first interview and aged 25-64 years at the follow up. MAIN RESULTS: Persons with limiting longstanding illness had a higher risk of adverse social consequences than persons without illness. The effect was modified by socioeconomic position only for labour market exclusion while the effects on unemployment and financial difficulties were equal across socioeconomic groups. CONCLUSIONS: Labour market policies as well as income maintenance policies that deal with social and economical consequences of longstanding illness are important elements of programmes to tackle inequalities in health. Rehabilitation within health care has a similar important part to play in this. PMID- 11854340 TI - Neighbourhood level versus individual level correlates of women's body dissatisfaction: toward a multilevel understanding of the role of affluence. AB - STUDY OBJECTIVE: This study examined the prevalence of body dissatisfaction as a function of individual level and neighbourhood level indicators of affluence. PARTICIPANTS AND SETTING: A subset of data from a larger random digit dialling telephone survey was used to obtain individual level data on body dissatisfaction, body weight and height, and income from a group of 895 adult women (age 24-56, 61% English speaking) living in 52 neighbourhoods (census tract areas) within the provinces of Alberta, Ontario, and Quebec, Canada who were selected for their heterogeneity in social class. DESIGN: Aggregated census tract data from 1996 were used to develop neighbourhood indicators of affluence. Using hierarchical linear modelling, body dissatisfaction (dichotomous) was examined as a function of individual body mass index, individual level affluence and neighbourhood level affluence. MAIN RESULTS: The impact of body mass index on body dissatisfaction depended on the level of neighbourhood affluence: an average body mass index was associated with higher likelihood of reporting body dissatisfaction in a neighbourhood of above average affluence (71% probability) than in a neighbourhood of average affluence (58% probability), independent of a woman's individual affluence (whether she was low income or not). CONCLUSION: It is concluded that a clearer understanding of the role of affluence on body dissatisfaction can be achieved by a joint examination of individual and neighbourhood level influences. PMID- 11854341 TI - Socioeconomic differences in children's use of physician services in the Nordic countries. AB - OBJECTIVE: To assess the relation between socioeconomic factors and the use of physician services among children and whether variations of the level of co payment are correlated with different levels of inequalities in health services use. DESIGN: Description of the socioeconomic differences in the use of health care using data from countrywide postal surveys to parents. SETTING: The five Nordic countries in 1996. SUBJECTS: Samples of 15 000 children aged 2-17 years: 3000 children at random, from the national registry in each country. MAIN OUTCOME MEASURE: Odds ratios of use of GP, specialist, and hospital services between children according to the educational level of both parents and the disposable income of the family, for all countries together and for each country separately. Odds ratios were adjusted for age, sex, urbanisation grade, and health status. RESULTS: There was little difference in the use of GP services according to socioeconomic factors. Parents from lower socioeconomic groups used telephone services of physicians less than parents from the higher groups and children of lower socioeconomic groups were seen less often by specialists. The reverse was true for hospitalisation of the children. The differential use of those three types of services was more marked in Denmark, Finland and Norway than in Iceland and Sweden. When controlled for other socioeconomic factors, the largest differences were observed according to the education of the mother. CONCLUSION: The specialist services and use of telephone services for children in the Nordic countries do not meet the criteria of equal use for equal need whereas the GP services and hospital services do to some extent. The education of the mother is a more important determinant than income for the use of each service. PMID- 11854342 TI - Long term mortality trends behind low life expectancy of Danish women. AB - STUDY OBJECTIVE: To examine the long time mortality trends of women in Denmark. DESIGN: Age-period-cohort analysis. SETTING: To search for possible causes we analysed mortality rates for all Danish women and men aged 40-84 during 1960 1999. Age-period-cohort modelling was used, handling the well known indeterminacy in a sensitivity analysis. MAIN RESULTS: The results indicate that the high risk of dying among Danish women is associated with being born between the two world wars. A similar pattern was not seen for men. CONCLUSION: The rather simple descriptive exploration in the framework of age-period-cohort modelling used, revealed a pattern not reflected by the commonly used life expectancy calculation. It is suggested that future studies on the low life expectancy of Danish women focus on the "between wars" generations identified with a high risk of dying in this study. PMID- 11854344 TI - Height, body mass index, and survival in men with coronary disease: follow up of the diet and reinfarction trial (DART). PMID- 11854345 TI - Balkans briefing number 7. Pristina Hospital: under new management. AB - This is the seventh paper in a series on the consequences for public health of the conflict in the Balkans. It describes the state of the hospital sevices in Pristina after the cease fire in June 1999 and the subsequent reconstruction programme. PMID- 11854343 TI - Does a higher number of siblings protect against the development of allergy and asthma? A review. AB - STUDY OBJECTIVE: To review the "protective" effects of having a higher number of siblings for the risk of atopic eczema, asthma wheezing, hay fever, and allergic sensitisation. METHOD: Review of the literature (Medline since 1965 and references). MAIN RESULTS: 53 different studies were identified. For eczema, 9 of 11 studies reported an inverse relation with number of siblings; for asthma and wheezing, 21 of 31 reported the inverse association; for hay fever, all 17 studies showed the effect; for allergic sensitisation or immunoglobulin E reactivity 14 of 16 studies supported the "protective" effect of a higher number of siblings. The studies emphasise a "theory" that is based exclusively on epidemiological associations. CONCLUSIONS: Research has not yet answered the question of which causal factors explain the sibling effect. Causal factors must meet two criteria; they must vary with sibship size and they must protect against atopic manifestations. The prevailing "hygiene hypothesis" failed to explain the findings adequately. Alternative explanations include in utero programming or endocrine explanatory models. The epidemiology research into siblings and atopic disorders has entered an intellectually challenging phase. Possessing sufficient knowledge about the causal factors might prevent at least 30% of all cases of asthma, eczema, and hay fever. PMID- 11854346 TI - Provision of services to asylum seekers. Are there lessons from the experience with Kosovan refugees? AB - Arrangements that were made to receive and support Kosovan refugees in the United Kingdom should be applied to asylum seekers arriving in the country. PMID- 11854347 TI - Smoking in young adolescents: an approach with multilevel discrete choice models. AB - STUDY OBJECTIVE: To understand the context for tobacco smoking in young adolescents, estimating the effects of individual, family, social, and school related factors. DESIGN: Cross sectional analysis performed by multilevel logistic regression with pupils at the first level and schools at the second level. The data came from a stratified sample of students surveyed on their own, their families' and their friends' smoking habits, their schools, and their awareness of cigarette prices and advertising. SETTING: The study was performed in the Island of Gran Canaria, Spain. PARTICIPANTS: 1877 students from 30 secondary schools in spring of 2000 (model's effective sample sizes 1697 and 1738). MAIN RESULTS: 14.2% of the young teenagers surveyed use tobacco, almost half of them (6.3% of the total surveyed) on a daily basis. According to the ordered logistic regression model, to have a smoker as the best friend increases significantly the probability of smoking (odds ratio: 6.96, 95% confidence intervals (CI) (4.93 to 9.84), and the same stands for one smoker living at home compared with a smoking free home (odds ratio: 2.03, 95% CI 1.22 to 3.36). Girls smoke more (odds ratio: 1.85, 95% CI 1.33 to 2.59). Experience with alcohol, and lack of interest in studies are also significant factors affecting smoking. Multilevel models of logistic regression showed that factors related to the school affect the smoking behaviour of young teenagers. More specifically, whether a school complies with antismoking rules or not is the main factor to predict smoking prevalence in schools. The remainder of the differences can be attributed to individual and family characteristics, tobacco consumption by parents or other close relatives, and peer group. CONCLUSIONS: A great deal of the individual differences in smoking are explained by factors at the school level, therefore the context is very relevant in this case. The most relevant predictors for smoking in young adolescents include some factors related to the schools they attend. One variable stood out in accounting for the school to school differences: how well they enforced the no smoking rule. Therefore we can prevent or delay tobacco smoking in adolescents not only by publicising health risks, but also by better enforcing no smoking rules in schools. PMID- 11854348 TI - Differential effect of socioeconomic status on rates of invasive coronary procedures across the public and private sectors in Queensland, Australia. PMID- 11854350 TI - Abortion and breast cancer. PMID- 11854349 TI - Re-emerging syphilis in gay men: a case-control study of behavioural risk factors and HIV status. PMID- 11854351 TI - Suicide in doctors. PMID- 11854352 TI - Description of a fitness to travel health assessment evaluation tool for displaced Albanian Kosovars in the former Yugoslav Republic of Macedonia. PMID- 11854353 TI - The production of a new MAGE-3 peptide presented to cytolytic T lymphocytes by HLA-B40 requires the immunoproteasome. AB - By stimulating human CD8(+) T lymphocytes with autologous dendritic cells infected with an adenovirus encoding MAGE-3, we obtained a cytotoxic T lymphocyte (CTL) clone that recognized a new MAGE-3 antigenic peptide, AELVHFLLL, which is presented by HLA-B40. This peptide is also encoded by MAGE-12. The CTL clone recognized MAGE-3--expressing tumor cells only when they were first treated with IFN-gamma. Since this treatment is known to induce the exchange of the three catalytic subunits of the proteasome to form the immunoproteasome, this result suggested that the processing of this MAGE-3 peptide required the immunoproteasome. Transfection experiments showed that the substitution of beta5i (LMP7) for beta5 is necessary and sufficient for producing the peptide, whereas a mutated form of beta5i (LMP7) lacking the catalytically active site was ineffective. Mass spectrometric analyses of in vitro digestions of a long precursor peptide with either proteasome type showed that the immunoproteasome produced the antigenic peptide more efficiently, whereas the standard proteasome more efficiently introduced cleavages destroying the antigenic peptide. This is the first example of a tumor-specific antigen exclusively presented by tumor cells expressing the immunoproteasome. PMID- 11854354 TI - A Trypanosoma cruzi small surface molecule provides the first immunological evidence that Chagas' disease is due to a single parasite lineage. AB - Chagas' disease is a major health and economic problem caused by the protozoan Trypanosoma cruzi. Multiple independently evolving clones define a complex parasite population that can be arranged into two broad genetic lineages termed T. cruzi I and II. These lineages have different evolutionary origin and display distinct ecological and biological traits. Here we describe a novel molecule termed TSSA for trypomastigote small surface antigen that provides the first immunological marker allowing discrimination between lineages. TSSA is a surface, glycosylphosphatidyl inositol (GPI)-anchored mucin-like protein, highly antigenic during the infection. TSSA sequences from different parasite isolates reveal a population dimorphism that perfectly matches with the two T. cruzi lineages. Interestingly, this dimorphism is restricted to the central region of the molecule, which comprises the immunodominant B cell epitopes. This sequence variability has a major impact on TSSA antigenicity, leading to no immunological cross-reactivity between both isoforms for antibodies present either in immunization or infection sera. Furthermore, the absolute seroprevalence for TSSA in confirmed Chagasic patients is restricted to T. cruzi II isoform, strongly suggesting that human infections are due to this particular subgroup. Even though association of T. cruzi II with Chagas' disease has been proposed based on molecular markers, this is the first immunological evidence supporting this hypothesis. The implications of these results for the future research on Chagas' disease could be envisaged. PMID- 11854355 TI - An immune evasion mechanism for spirochetal persistence in Lyme borreliosis. AB - Borrelia burgdorferi, the Lyme disease spirochete, persistently infects mammalian hosts despite the development of strong humoral responses directed against the pathogen. Here we describe a novel mechanism of immune evasion by B. burgdorferi. In immunocompetent mice, spirochetes that did not express ospC (the outer-surface protein C gene) were selected within 17 d after inoculation, concomitantly with the emergence of anti-OspC antibody. Spirochetes with no detectable OspC transcript that were isolated from immunocompetent mice reexpressed ospC after they were either cultured in vitro or transplanted to naive immunocompetent mice, but not in OspC-immunized mice. B. burgdorferi persistently expressed ospC in severe combined immune-deficient (SCID) mice. Passive immunization of B. burgdorferi-infected SCID mice with an anti-OspC monoclonal antibody selectively eliminated ospC-expressing spirochetes but did not clear the infection. OspC expressing spirochetes reappeared in SCID mice after the anti-OspC antibody was eliminated. We submit that selection of surface-antigen nonexpressers is an immune evasion mechanism that contributes to spirochetal persistence. PMID- 11854357 TI - Characterization of T cell differentiation in the murine gut. AB - Gut intraepithelial CD8 T lymphocytes (T-IEL) are distinct from thymus-derived cells and are thought to derive locally from cryptopatch (CP) precursors. The intermediate stages of differentiation between CP and mature T-IEL were not identified, and the local differentiation process was not characterized. We identified and characterized six phenotypically distinct lineage-negative populations in the CP and the gut epithelium: (a) we determined the kinetics of their generation from bone marrow precursors; (b) we quantified CD3-epsilon, recombination activating gene (Rag)-1, and pre-Talpha mRNAs expression at single cell level; (c) we characterized TCR-beta, -gamma, and -alpha locus rearrangements; and (d) we studied the impact of different mutations on the local differentiation. These data allowed us to establish a sequence of T cell precursor differentiation in the gut. We also observed that the gut differentiation varied from that of the thymus by a very low frequency of pre Talpha chain mRNA expression, a different kinetics of Rag-1 mRNA expression, and a much higher impact of CD3 epsilon/delta and pre-Talpha deficiencies. Finally, only 3% of CP cells were clearly involved in T cell differentiation, suggesting that these structures may have additional physiological roles in the gut. PMID- 11854356 TI - Tumor growth enhances cross-presentation leading to limited T cell activation without tolerance. AB - Using a tumor model of spontaneously arising insulinomas expressing a defined tumor-associated antigen, we investigated whether tumor growth promotes cross presentation and tolerance of tumor-specific T cells. We found that an advanced tumor burden enhanced cross-presentation of tumor-associated antigens to high avidity tumor-specific T cells, inducing T cell proliferation and limited effector function in vivo. However, contrary to other models, tumor-specific T cells were not tolerized despite a high tumor burden. In fact, in tumor-bearing mice, persistence and responsiveness of adoptively transferred tumor-specific T cells were enhanced. Accordingly, a potent T cell-mediated antitumor response could be elicited by intravenous administration of tumor-derived peptide and agonistic anti-CD40 antibody or viral immunization and reimmunization. Thus, in this model, tumor growth promotes activation of high avidity tumor-specific T cells instead of tolerance. Therefore, the host remains responsive to T cell immunotherapy. PMID- 11854358 TI - Complement interaction with trypanosomatid promastigotes in normal human serum. AB - In normal human serum (NHS), axenic promastigotes of Crithidia, Phytomonas, and Leishmania trigger complement activation, and from 1.2 to 1.8 x 10(5) C3 molecules are deposited per promastigote within 2.5 min. In Leishmania, promastigote C3 binding capacity remains constant during in vitro metacyclogenesis. C3 deposition on promastigotes activated through the classical complement pathway reaches a 50% maximum after similar50 s, and represents >85% of total C3 bound. In C1q- and C2-deficient human sera, promastigotes cannot activate the classical pathway (CP) unless purified C1q or C2 factors, respectively, are supplemented, demonstrating a requirement for CP factor in promastigote C3 opsonization. NHS depleted of natural anti-Leishmania antibodies cannot trigger promastigote CP activation, but IgM addition restores C3 binding. Furthermore, Leishmania binds natural antibodies in ethylenediaminetetracetic acid (EDTA)-treated NHS; after EDTA removal, promastigote-bound IgM triggers C3 deposition in natural antibody-depleted NHS. Serum collectins and pentraxins thus do not participate significantly in NHS promastigote C3 opsonization. Real-time kinetic analysis of promastigote CP-mediated lysis indicates that between 85--95% of parasites are killed within 2.5 min of serum contact. These data indicate that successful Leishmania infection in man must immediately follow promastigote transmission, and that Leishmania evasion strategies are shaped by the selective pressure exerted by complement. PMID- 11854359 TI - Dynamics of major histocompatibility complex class II compartments during B cell receptor-mediated cell activation. AB - Antigen recognition by clonotypic B cell receptor (BcR) is the first step of B lymphocytes differentiation into plasmocytes. This B cell function is dependent on efficient major histocompatibility complex (MHC) class II-restricted presentation of BcR-bound antigens. In this work, we analyzed the subcellular mechanisms underlying antigen presentation after BcR engagement on B cells. In quiescent B cells, we found that MHC class II molecules mostly accumulated at the cell surface and in an intracellular pool of tubulovesicular structures, whereas H2-M molecules were mostly detected in distinct lysosomal compartments devoid of MHC class II. BcR stimulation induced the transient intracellular accumulation of MHC class II molecules in newly formed multivesicular bodies (MVBs), to which H2 M was recruited. The reversible downregulation of cathepsin S activity led to the transient accumulation of invariant chain-MHC class II complexes in MVBs. A few hours after BcR engagement, cathepsin S activity increased, the p10 invariant chain disappeared, and MHC class II-peptide complexes arrived at the plasma membrane. Thus, BcR engagement induced the transient formation of antigen processing compartments, enabling antigen-specific B cells to become effective antigen-presenting cells. PMID- 11854360 TI - Complementary dendritic cell-activating function of CD8+ and CD4+ T cells: helper role of CD8+ T cells in the development of T helper type 1 responses. AB - Dendritic cells (DCs) activated by CD40L-expressing CD4+ T cells act as mediators of "T helper (Th)" signals for CD8+ T lymphocytes, inducing their cytotoxic function and supporting their long-term activity. Here, we show that the optimal activation of DCs, their ability to produce high levels of bioactive interleukin (IL)-12p70 and to induce Th1-type CD4+ T cells, is supported by the complementary DC-activating signals from both CD4+ and CD8+ T cells. Cord blood- or peripheral blood-isolated naive CD8+ T cells do not express CD40L, but, in contrast to naive CD4+ T cells, they are efficient producers of IFN-gamma at the earliest stages of the interaction with DCs. Naive CD8+ T cells cooperate with CD40L-expressing naive CD4+ T cells in the induction of IL-12p70 in DCs, promoting the development of primary Th1-type CD4+ T cell responses. Moreover, the recognition of major histocompatibility complex class I-presented epitopes by antigen-specific CD8+ T cells results in the TNF-alpha- and IFN-gamma-dependent increase in the activation level of DCs and in the induction of type-1 polarized mature DCs capable of producing high levels of IL-12p70 upon a subsequent CD40 ligation. The ability of class I-restricted CD8+ T cells to coactivate and polarize DCs may support the induction of Th1-type responses against class I-presented epitopes of intracellular pathogens and contact allergens, and may have therapeutical implications in cancer and chronic infections. PMID- 11854361 TI - Thymic selection generates a large T cell pool recognizing a self-peptide in humans. AB - The low frequency of self-peptide-specific T cells in the human preimmune repertoire has so far precluded their direct evaluation. Here, we report an unexpected high frequency of T cells specific for the self-antigen Melan-A/MART-1 in CD8 single-positive thymocytes from human histocompatibility leukocyte antigen A2 healthy individuals, which is maintained in the peripheral blood of newborns and adults. Postthymic replicative history of Melan-A/MART-1-specific CD8 T cells was independently assessed by quantifying T cell receptor excision circles and telomere length ex vivo. We provide direct evidence that the large T cell pool specific for the self-antigen Melan-A/MART-1 is mostly generated by thymic output of a high number of precursors. This represents the only known naive self-peptide specific T cell repertoire directly accessible in humans. PMID- 11854362 TI - Maximal HIV-1 replication in alveolar macrophages during tuberculosis requires both lymphocyte contact and cytokines. AB - HIV-1 replication is markedly upregulated in alveolar macrophages (AM) during pulmonary tuberculosis (TB). This is associated with loss of an inhibitory CCAAT enhancer binding protein beta (C/EBPbeta) transcription factor and activation of nuclear factor (NF)-kappaB. Since the cellular immune response in pulmonary TB requires lymphocyte--macrophage interaction, a model system was developed in which lymphocytes were added to AM. Contact between lymphocytes and AM reduced inhibitory C/EBPbeta, activated NF-kappaB, and enhanced HIV-1 replication. If contact between lymphocytes and macrophages was prevented, inhibitory C/EBPbeta expression was maintained and the HIV-1 long terminal repeat (LTR) was not maximally stimulated although NF-kappaB was activated. Antibodies that cross linked macrophage expressed B-7, and vascular cell adhesion molecule and CD40 were used to mimic lymphocyte contact. All three cross-linking antibodies were required to abolish inhibitory C/EBPbeta expression. However, the HIV-1 LTR was not maximally stimulated and NF-kappaB was not activated. Maximal HIV-1--LTR stimulation required both lymphocyte-derived soluble factors, and cross-linking of macrophage expressed costimulatory molecules. High level HIV-1--LTR stimulation was also achieved when IL-1beta, IL-6, and TNF-beta were added to macrophages with cross-linked costimulatory molecules. Contact between activated lymphocytes and macrophages is necessary to down-regulate inhibitory C/EBPbeta, thereby derepressing the HIV-1 LTR. Lymphocyte-derived cytokines activate NF kappaB, further enhancing the HIV-1 LTR. PMID- 11854363 TI - Virus-induced interferon alpha production by a dendritic cell subset in the absence of feedback signaling in vivo. AB - An effective type I interferon (IFN-alpha/beta) response is critical for the control of many viral infections. Here we show that in vesicular stomatitis virus (VSV)-infected mouse embryonic fibroblasts (MEFs) the production of IFN-alpha is dependent on type I IFN receptor (IFNAR) triggering, whereas in infected mice early IFN-alpha production is IFNAR independent. In VSV-infected mice type I IFN is produced by few cells located in the marginal zone of the spleen. Unlike other dendritic cell (DC) subsets, FACS((R))-sorted CD11c(int)CD11b(-)GR-1(+) DCs show high IFN-alpha expression, irrespective of whether they were isolated from VSV infected IFNAR-competent or -deficient mice. Thus, VSV preferentially activates a specialized DC subset presumably located in the marginal zone to produce high level IFN-alpha largely independent of IFNAR feedback signaling. PMID- 11854364 TI - Interferon alpha/beta and interleukin 12 responses to viral infections: pathways regulating dendritic cell cytokine expression in vivo. AB - Interferon (IFN)-alpha/beta and interleukin (IL)-12 are cytokines critical in defense against viruses, but their cellular sources and mechanisms of regulation for in vivo expression remain poorly characterized. The studies presented here identified a novel subset of dendritic cells (DCs) as major producers of the cytokines during murine cytomegalovirus (MCMV) but not lymphocytic choriomeningitis virus (LCMV) infections. These DCs differed from those activated by Toxoplasma antigen but were related to plasmacytoid cells, as assessed by their CD8alpha(+)Ly6G/C(+)CD11b(-) phenotype. Another DC subset (CD8alpha(2)Ly6G/C(-)CD11b(+)) also contributed to IL-12 production in MCMV infected immunocompetent mice, modestly. However, it dramatically increased IL-12 expression in the absence of IFN-alpha/beta functions. Conversely, IFN-alpha/beta production was greatly reduced under these conditions. Thus, a cross-regulation of DC subset cytokine responses was defined, whereby secretion of type I IFNs by CD8alpha(+) DCs resulted in responses limiting IL-12 expression by CD11b(+) DCs but enhancing overall IFN-alpha/beta production. Taken together, these data indicate that CD8alpha(+)Ly6G/C(+)CD11b(-) DCs play important roles in limiting viral replication and regulating immune responses, through cytokine production, in some but not all viral infections. They also illustrate the plasticity of cellular sources for innate cytokines in vivo and provide new insights into the roles of IFNs in shaping immune responses to viruses. PMID- 11854365 TI - Activation-induced deaminase (AID)-directed hypermutation in the immunoglobulin Smu region: implication of AID involvement in a common step of class switch recombination and somatic hypermutation. AB - Somatic hypermutation (SHM) and class switch recombination (CSR) cause distinct genetic alterations at different regions of immunoglobulin genes in B lymphocytes: point mutations in variable regions and large deletions in S regions, respectively. Yet both depend on activation-induced deaminase (AID), the function of which in the two reactions has been an enigma. Here we report that B cell stimulation which induces CSR but not SHM, leads to AID-dependent accumulation of SHM-like point mutations in the switch mu region, uncoupled with CSR. These findings strongly suggest that AID itself or a single molecule generated by RNA editing function of AID may mediate a common step of SHM and CSR, which is likely to be involved in DNA cleavage. PMID- 11854367 TI - Trimethylamine oxide accumulation in marine animals: relationship to acylglycerol storage. AB - Trimethylamine oxide (TMAO) is a common and compatible osmolyte in muscle tissues of marine organisms that is often credited with counteracting protein destabilizing forces. However, the origin and synthetic pathways of TMAO are actively debated. Here, we examine the distribution of TMAO in marine animals and report a correlation between TMAO and acylglycerol storage. We put forward the hypothesis that TMAO is derived, at least in part, from the hydrolysis of phosphatidylcholine, endogenous or dietary, for storage as diacylglycerol ethers and triacylglycerols. TMAO is synthesized from the trimethylammonium moiety of choline, thus released, and is retained as a compatible solute in concentrations reflecting the amount of lipid stored in the body. A variation on this theme is proposed for sharks. PMID- 11854366 TI - Whence interferon? Variety in the production of interferon in response to viral infection. PMID- 11854368 TI - Implanted electrode recordings from a praying mantis auditory interneuron during flying bat attacks. AB - Using an implanted electrode, we recorded the responses from the ultrasound sensitive mantis interneuron 501-T3 during flying bat attacks in a large flight room where the mantis served as the target. 501-T3 responds to each vocalization emitted with multi-spike bursts when pulse repetition rates (PRRs) are below 55 pulses x s(-1). As PRR increases and pulse durations fall below 3 ms, 501-T3 ceases burst activity. On average, spike bursts cease 272 ms before contact (when the bat is 73 cm away from the preparation). The timing of cessation of activity in 501-T3 is similar to the latency for the diving portion of the response of the mantid (242 ms). Experiments using vocalizing stationary bats confirm that 501-T3 responds more reliably to longer pulse durations (> or =3 ms) when intensities are below 90 dB pe SPL. The cessation of 501-T3 activity is probably due both to the increasing PRR and to the decreasing pulse duration that occur in the terminal buzz phase of a bat attack. 501-T3 may be actively shut off at high PRRs and/or intensities to protect the interneuron from habituation while the mantis performs an escape response. The cessation of 501-T3 activity is consistent with the lack of a very late ultrasound-mediated evasive response by the mantis. However, cessation of 501-T3 activity may allow a true 'last-chance' response to be mediated by other neural systems. PMID- 11854370 TI - The influence of visual landscape on the free flight behavior of the fruit fly Drosophila melanogaster. AB - To study the visual cues that control steering behavior in the fruit fly Drosophila melanogaster, we reconstructed three-dimensional trajectories from images taken by stereo infrared video cameras during free flight within structured visual landscapes. Flies move through their environment using a series of straight flight segments separated by rapid turns, termed saccades, during which the fly alters course by approximately 90 degrees in less than 100 ms. Altering the amount of background visual contrast caused significant changes in the fly's translational velocity and saccade frequency. Between saccades, asymmetries in the estimates of optic flow induce gradual turns away from the side experiencing a greater motion stimulus, a behavior opposite to that predicted by a flight control model based upon optomotor equilibrium. To determine which features of visual motion trigger saccades, we reconstructed the visual environment from the fly's perspective for each position in the flight trajectory. From these reconstructions, we modeled the fly's estimation of optic flow on the basis of a two-dimensional array of Hassenstein-Reichardt elementary motion detectors and, through spatial summation, the large-field motion stimuli experienced by the fly during the course of its flight. Event-triggered averages of the large-field motion preceding each saccade suggest that image expansion is the signal that triggers each saccade. The asymmetry in output of the local motion detector array prior to each saccade influences the direction (left versus right) but not the magnitude of the rapid turn. Once initiated, visual feedback does not appear to influence saccade kinematics further. The total expansion experienced before a saccade was similar for flight within both uniform and visually textured backgrounds. In summary, our data suggest that complex behavioral patterns seen during free flight emerge from interactions between the flight control system and the visual environment. PMID- 11854369 TI - Evidence for increased myofibrillar mobility in desmin-null mouse skeletal muscle. AB - Quantitative electron microscopy was used to characterize the longitudinal mobility of myofibrils during muscle extension to investigate the functional roles of skeletal muscle intermediate filaments. Extensor digitorum longus fifth toe muscles from wild-type (+/+) and desmin-null (des -/-) animals were passively stretched to varying lengths, and the horizontal displacement of adjacent Z-disks in neighboring myofibrils (Deltax(myo)) and average sarcomere length (SL) were calculated. At short SL (<2.20 microm), wild-type and desmin-null Deltax(myo) were not significantly different, although there was a trend towards greater Z disk misalignment in muscles from knockout animals (Deltax(myo) 0.34+/-0.04 microm versus 0.22+/-0.09 microm; P>0.2; means +/- S.E.M.). However, at higher SL (>2.90 microm), muscles from knockout animals displayed a dramatically increased Deltax(myo) relative to wild-type muscles (0.49+/-0.10 microm versus 0.25+/-0.07 microm; P<0.05). The results, which establish a maximum extension of the desmin network surrounding the Z-disk, provide what we believe to be the first quantitative estimation of the functional limits of the desmin intermediate filament system in the presence of an intact myofibrillar lattice. The existence of a limit on the extension of desmin suggests a mechanism for the recruitment of desmin into a network of force transmission, whether as a longitudinal load bearer or as a component in a radial force-transmission system. PMID- 11854371 TI - Tissue- and development-specific induction and turnover of hsp70 transcripts from loci 87A and 87C after heat shock and during recovery in Drosophila melanogaster. AB - The haploid genome of Drosophila melanogaster normally carries at least five nearly identical copies of heat-shock-inducible hsp70 genes, two copies at the 87A7 and three copies at the 87C1 chromosome sites. We used in situ hybridization of the cDNA, which hybridizes with transcripts of all five hsp70 genes, and of two 3' untranslated region (3'UTR; specific for the 87A7- and 87C1-type hsp70 transcripts) riboprobes to cellular RNA to examine whether all these copies were similarly induced by heat shock in different cell types of D. melanogaster. Our results revealed remarkable differences not only in the heat-shock-inducibility of the hsp70 genes at the 87A7 and 87C1 loci, but also in their post transcriptional metabolism, such as the stability of the transcripts and of their 3'UTRs in different cell types in developing embryos and in larval and adult tissues. Our results also revealed the constitutive presence of the heat-shock inducible form of Hsp70 in a subset of late spermatogonial cells from the second instar larval stage onwards. We suggest that the multiple copies of the stress inducible hsp70 genes do not exist in the genome of D. melanogaster only to produce large amounts of the Hsp70 rapidly and at short notice, but that they are specifically regulated in a developmental-stage-specific manner. It is likely that the cost/benefit ratio of not producing or of producing a defined amount of Hsp70 under stress conditions varies for different cell types and under different physiological conditions and, accordingly, specific regulatory mechanisms operating at the transcriptional and post-transcriptional levels have evolved. PMID- 11854372 TI - Determining patterns of motor recruitment during locomotion. AB - Motor units are the functional units of muscle contraction in vertebrates. Each motor unit comprises muscle fibres of a particular fibre type and can be considered as fast or slow depending on its fibre-type composition. Motor units are typically recruited in a set order, from slow to fast, in response to the force requirements from the muscle. The anatomical separation of fast and slow muscle in fish permits direct recordings from these two fibre types. The frequency spectra from different slow and fast myotomal muscles were measured in the rainbow trout Oncorhynchus mykiss. These two muscle fibre types generated distinct low and high myoelectric frequency bands. The cat paw-shake is an activity that recruits mainly fast muscle. This study showed that the myoelectric signal from the medial gastrocnemius of the cat was concentrated in a high frequency band during paw-shake behaviour. During slow walking, the slow motor units of the medial gastrocnemius are also recruited, and this appeared as increased muscle activity within a low frequency band. Therefore, high and low frequency bands could be distinguished in the myoelectric signals from the cat medial gastrocnemius and probably corresponded, respectively, to fast and slow motor unit recruitment. Myoelectric signals are resolved into time/frequency space using wavelets to demonstrate how patterns of motor unit recruitment can be determined for a range of locomotor activities. PMID- 11854373 TI - Kinematics of diving Atlantic puffins (Fratercula arctica L.): evidence for an active upstroke. AB - To examine the propulsion mechanism of diving Atlantic puffins (Fratercula arctica), their three-dimensional kinematics was investigated by digital analysis of sequential video images of dorsal and lateral views. During the dives of this wing-propelled bird, the wings are partly folded, with the handwings directed backwards. The wings go through an oscillating motion in which the joint between the radius-ulna and the hand bones leads the motion, with the wing tip following. There is a large rotary motion of the wings during the stroke, with the wings being pronated at the beginning of the downstroke and supinated at the end of the downstroke/beginning of the upstroke. Calculated instantaneous velocities and accelerations of the bodies of the birds show that, during the downstroke, the birds accelerate upwards and forwards. During the upstroke, the birds accelerate downwards and, in some sequences analysed, also forwards, but in most cases the birds decelerate. In all the upstrokes analysed, the forward/backward acceleration shows the same pattern, with a reduced deceleration or even a forward acceleration during 'mid' upstroke indicating the production of a forward force, thrust. Our results show that the Atlantic puffin can use an active upstroke during diving, in contradiction to previous data. Furthermore, we suggest that the partly folded wings of diving puffins might act as efficient aft swept wingtips, reducing the induced drag and increasing the lift-to-drag ratio. A movie is available on-line. PMID- 11854374 TI - A motor and a brake: two leg extensor muscles acting at the same joint manage energy differently in a running insect. AB - The individual muscles of a multiple muscle group at a given joint are often assumed to function synergistically to share the load during locomotion. We examined two leg extensors of a running cockroach to test the hypothesis that leg muscles within an anatomical muscle group necessarily manage (i.e. produce, store, transmit or absorb) energy similarly during running. Using electromyographic and video motion-analysis techniques, we determined that muscles 177c and 179 are both active during the first half of the stance period during muscle shortening. Using the in vivo strain and stimulation patterns determined during running, we measured muscle power output. Although both muscles were stimulated during the first half of shortening, muscle 177c generated mechanical energy (28 W x kg(-1)) like a motor, while muscle 179 absorbed energy (-19 W x kg(-1)) like a brake. Both muscles exhibited nearly identical intrinsic characteristics including similar twitch kinetics and force-velocity relationships. Differences in the extrinsic factors of activation and relative shortening velocity caused the muscles to operate very differently during running. Presumed redundancy in a multiple muscle group may, therefore, represent diversity in muscle function. Discovering how muscles manage energy during behavior requires the measurement of a large number of dynamically interacting variables. PMID- 11854375 TI - Differences in behaviour between rainbow trout selected for high- and low-stress responsiveness. AB - Two F1 lines of rainbow trout Oncorhynchus mykiss, divergent for plasma cortisol responsiveness, were generated by individual selection for post-stress cortisol values within the F0 generation. Adult females of the F1 generation were transferred to rearing in social isolation in observation tanks. After 6 days, locomotor activity in high-responding (HR) and low-responding (LR) individuals was quantified as time spent moving during a 20 min observation period. Behavioural observations were repeated the next day with a smaller conspecific intruder present in each observation tank. Differential hypothalamus-pituitary interrenal axis activity in the two lines was subsequently confirmed by a standardised confinement stress test, which resulted in significantly higher plasma cortisol concentrations in HR than LR fish. HR fish displayed higher levels of locomotor activity than LR fish in the presence of an intruder, but not when in isolation. Aggressive behaviour towards the intruder was not seen, suggesting either a state-dependent lack of territorial aggression, or chronic stress in the experimental fish. A significantly higher incidence of feed intake was seen in LR trout when held in observation tanks (40% versus 0% of the fish took food when in isolation), suggesting that these fish acclimated more successfully to the experimental conditions than HR fish did. These results suggest that selection for stress responsiveness in salmonid fish leads to behavioural alterations, which are of potential importance to the performance of these fish in aquaculture rearing operations. PMID- 11854377 TI - Copper transport by lobster (Homarus americanus) hepatopancreatic mitochondria. AB - Mechanisms of copper transport into purified mitochondrial suspensions prepared from the hepatopancreas of the Atlantic lobster Homarus americanus were investigated. Mitochondria were purified by combining methods of differential and Percoll-gradient centrifugation, and copper transport was studied using the copper-sensitive fluorescent dye Phen Green. Copper transport by this mitochondrial preparation was kinetically the sum of saturable and non-saturable transfer components. Addition of 500 micromol x l(-1) Ca2+ or 500 nmol x l(-1) Ruthenium Red abolished the non-saturable copper transport component, significantly (P<0.01) reduced the apparent binding affinity of the saturable transport component, but was without effect (P>0.05) on the apparent maximal transport velocity of the saturable transfer process. The antiport inhibitor diltiazem (500 micromol x l(-1)) acted as a mixed inhibitor of the saturable transport mechanism, but had no effect on the non-saturable component of transfer. These results suggest that the non-saturable copper influx process was probably by way of the well-known Ruthenium-Red-sensitive Ca2+ uniporter and that the saturable transport component was probably due to a combination of both the Na+-dependent, diltiazem-sensitive 1Ca2+/2Na+ antiporter and the Na+-independent, diltiazem-insensitive 1Ca2+/2H+ antiporter. A model is discussed relating these mitochondrial copper uptake processes to the transfer of metal ions across the epithelial brush-border membrane. PMID- 11854376 TI - Modulation of swimming in the gastropod Melibe leonina by nitric oxide. AB - Nitric oxide (NO) is a gaseous intercellular messenger produced by the enzyme nitric oxide synthase. It has been implicated as a neuromodulator in several groups of animals, including gastropods, crustaceans and mammals. In this study, we investigated the effects of NO on the swim motor program produced by isolated brains and by semi-intact preparations of the nudibranch Melibe leonina. The NO donors sodium nitroprusside (SNP, 1 mmol x l(-1)) and S-nitroso-N acetylpenicillamine (SNAP, 1 mmol x l(-1)) both had a marked effect on the swim motor program expressed in isolated brains, causing an increase in the period of the swim cycle and a more erratic swim rhythm. In semi-intact preparations, the effect of NO donors was manifested as a significant decrease in the rate of actual swimming. An NO scavenger, reduced oxyhemoglobin, eliminated the effects of NO donors on isolated brains, supporting the assumption that the changes in swimming induced by donors were actually due to NO. The cGMP analogue 8 bromoguanosine 3',5'-cyclic monophosphate (1 mmol x l(-1)) produced effects that mimicked those of NO donors, suggesting that NO is working via a cGMP-dependent mechanism. These results, in combination with previous histological studies indicating the endogenous presence of nitric oxide synthase, suggest that NO is used in the central nervous system of Melibe leonina to modulate swimming. PMID- 11854378 TI - The pulmonary surfactant system matures upon pipping in the freshwater turtle Chelydra serpentina. AB - Pulmonary surfactant (PS), a mixture of phospholipids (PL), neutral lipids and surfactant proteins (SP), lowers surface tension within the lung, which increases lung compliance and improves the removal of fluid at birth. Here, we have examined the expression of thyroid transcription factor-1 (TTF-1) and the surfactant protein SP-B, and also the composition of pulmonary surfactant lipids in the developing lung of the turtle Chelydra serpentina. Lavage and lung tissue were collected from late embryonic, pipped and hatchling turtles. TTF-1, a regulator of gene expression of surfactant proteins and cell differentiation in mammals, was detected using immunohistochemistry in epithelia of the gas-exchange area and conducting airways during late development. Expression declined in hatchlings. SP-B was detected in subsets of cells within the respiratory epithelium at all stages sampled. The same cell types also stained for TTF-1. Turtle surfactant lipids matured toward the end of incubation. Maximal secretion of both total phospholipids and disaturated phospholipid (DSP) occurred at the time of pipping, coincident with the onset of breathing. The DSP/PL ratio increased after pipping, whereas cholesterol levels (Chol) increased prior to pipping. This resulted in a decrease in the Chol/PL and Chol/DSP ratios after pipping. Thus, TTF-1 and SP-B appear to be highly conserved within the vertebrates. Maturation of surfactant phospholipid content occurred with the commencement of pulmonary ventilation. PMID- 11854379 TI - Morphology, swimming performance and propulsive mode of six co-occurring hydromedusae. AB - Jet propulsion, based on examples from the Hydrozoa, has served as a valuable model for swimming by medusae. However, cnidarian medusae span several taxonomic classes (collectively known as the Medusazoa) and represent a diverse array of morphologies and swimming styles. Does one mode of propulsion appropriately describe swimming by all medusae? This study examined a group of co-occurring hydromedusae collected from the waters of Friday Harbor, WA, USA, to investigate relationships between swimming performance and underlying mechanisms of thrust production. The six species examined encompassed a wide range of bell morphologies and swimming habits. Swimming performance (measured as swimming acceleration and velocity) varied widely among the species and was positively correlated with bell streamlining (measured as bell fineness ratio) and velar structure development (measured as velar aperture ratio). Calculated thrust production due to jet propulsion adequately explained acceleration patterns of prolate medusae (Aglantha digitale, Sarsia sp. and Proboscidactyla flavicirrata) possessing well-developed velums. However, acceleration patterns of oblate medusae (Aequorea victoria, Mitrocoma cellularia and Phialidium gregarium) that have less developed velums were poorly described by jet thrust production. An examination of the wakes behind swimming medusae indicated that, in contrast to the clearly defined jet structures produced by prolate species, oblate medusae did not produce defined jets but instead produced prominent vortices at the bell margins. These vortices are consistent with a predominantly drag-based, rowing mode of propulsion by the oblate species. These patterns of propulsive mechanics and swimming performance relate to the role played by swimming in the foraging ecology of each medusa. These patterns appear to extend beyond hydromedusae and thus have important implications for other members of the Medusazoa. PMID- 11854380 TI - Polyomavirus and medulloblastoma: a smoking gun or guilt by association? PMID- 11854381 TI - Hobson's choice and the need for combinations of new agents for the prevention and treatment of breast cancer. PMID- 11854382 TI - Mutation analysis suggests role for ribonuclease L gene in prostate cancer. PMID- 11854383 TI - Developing countries put cervical cancer tests to the test. PMID- 11854384 TI - Estimated cervical cancer mortality in selected countries, 2000. PMID- 11854385 TI - COX-2 inhibitors: cancer trials test new uses for pain drug. PMID- 11854386 TI - Insurers, employers ask for better evaluation of new technologies. PMID- 11854387 TI - Nonsteroidal anti-inflammatory drugs as anticancer agents: mechanistic, pharmacologic, and clinical issues. AB - Numerous experimental, epidemiologic, and clinical studies suggest that nonsteroidal anti-inflammatory drugs (NSAIDs), particularly the highly selective cyclooxygenase (COX)-2 inhibitors, have promise as anticancer agents. NSAIDs restore normal apoptosis in human adenomatous colorectal polyps and in various cancer cell lines that have lost adenomatous polyposis coli gene function. NSAIDs also inhibit angiogenesis in cell culture and rodent models of angiogenesis. Many epidemiologic studies have found that long-term use of NSAIDs is associated with a lower risk of colorectal cancer, adenomatous polyps, and, to some extent, other cancers. Two NSAIDs, sulindac and celecoxib, have been found to inhibit the growth of adenomatous polyps and cause regression of existing polyps in randomized trials of patients with familial adenomatous polyposis (FAP). However, unresolved questions about the safety, efficacy, optimal treatment regimen, and mechanism of action of NSAIDs currently limit their clinical application to the prevention of polyposis in FAP patients. Moreover, the development of safe and effective drugs for chemoprevention is complicated by the potential of even rare, serious toxicity to offset the benefit of treatment, particularly when the drug is administered to healthy people who have low annual risk of developing the disease for which treatment is intended. This review considers generic approaches to improve the balance between benefits and risks associated with the use of NSAIDs in chemoprevention. We critically examine the published experimental, clinical, and epidemiologic literature on NSAIDs and cancer, especially that regarding colorectal cancer, and identify strategies to overcome the various logistic and scientific barriers that impede clinical trials of NSAIDs for cancer prevention. Finally, we suggest research opportunities that may help to accelerate the future clinical application of NSAIDs for cancer prevention or treatment. PMID- 11854388 TI - Expression of human neurotropic polyomavirus JCV late gene product agnoprotein in human medulloblastoma. AB - BACKGROUND: The human neurotropic polyomavirus, JCV, contains an open reading frame within the late region of the viral genome that encodes a 71-amino-acid protein, agnoprotein. Because accumulating evidence supports an association between JCV infection and human brain tumors, including medulloblastomas, we assessed the presence of JCV Agno gene sequences and the expression of agnoprotein in a series of 20 well-characterized medulloblastomas. METHODS: Formalin-fixed, paraffin-embedded tumor tissue samples were used for Agno gene amplification and for immunohistochemical analysis. Adjacent sections were stained with an antibody to agnoprotein and with antibodies to cellular structural and regulatory proteins, including the JCV early gene product, T antigen. RESULTS: Analysis of amplified DNA from paraffin-embedded samples revealed the presence of the Agno gene in 11 (69%) of 16 samples. Immunohistochemical analysis showed cytoplasmic localization and widespread distribution of agnoprotein in the neoplastic cells in 11 (55%) of 20 samples. The JCV early gene product, T antigen, was present in the nucleus of some, but not all, of the neoplastic cells. Some medulloblastoma samples that expressed agnoprotein had no sign of T-antigen expression. p53 was detected in only six of the 11 tumors in which agnoprotein was expressed. None of the 20 samples showed expression of the viral late capsid proteins, ruling out productive infection of the tumor cells with JCV. CONCLUSIONS: Our data provide evidence that the JCV late gene encoding the auxiliary agnoprotein is expressed in tumor cells. The finding of agnoprotein expression in the absence of T-antigen expression suggests a potential role for agnoprotein in pathways involved in the development of JCV associated medulloblastomas. PMID- 11854389 TI - Effects of raloxifene after tamoxifen on breast and endometrial tumor growth in athymic mice. AB - BACKGROUND: In patients with early-stage breast cancer, 5 years of treatment with the selective estrogen receptor modulator (SERM) tamoxifen reduces breast cancer recurrence and mortality, whereas more than 5 years of tamoxifen does not further reduce breast cancer recurrence and doubles the risk of endometrial cancer. We evaluated the effects on tumor growth of raloxifene, another SERM, after tamoxifen treatment in mouse models of breast and endometrial cancers. METHODS: Athymic, ovariectomized mice were bitransplanted with tumors derived from human breast cancer and endometrial cancer cells that either were tamoxifen-naive or had been exposed to tamoxifen for short (6 months) or long (>5 years) terms. The effects of raloxifene (two dose levels) and tamoxifen on tumor growth in the presence and absence of low-dose estrogen were evaluated. All statistical tests were two-sided. RESULTS: Raloxifene was less effective than tamoxifen in blocking the stimulatory effects of low-dose estrogen on the growth of tamoxifen-naive breast (P<.001) and endometrial (P =.001) tumors. Raloxifene and tamoxifen had similar inhibitory effects on the growth of short-term tamoxifen-exposed breast tumors. Raloxifene and tamoxifen had similar stimulatory effects on the growth of breast and endometrial tumors that had been exposed to at least 5 years of tamoxifen. However, neither drug blocked the stimulatory effects of estrogen on the growth of these tumors. Raloxifene was less effective than tamoxifen (P<.001) in blocking the stimulatory effects of estrogen on endometrial tumors that had been exposed to tamoxifen in the past. CONCLUSIONS: Raloxifene and tamoxifen had similar effects on these mouse models of tamoxifen-naive and tamoxifen-resistant breast and endometrial cancer. Treatment with raloxifene following 5 years of adjuvant tamoxifen may not further decrease breast cancer recurrence and may increase endometrial cancer incidence. PMID- 11854390 TI - Outcomes among African-American/non-African-American patients with advanced non small-cell lung carcinoma: report from the Cancer and Leukemia Group B. AB - BACKGROUND: Among patients diagnosed with advanced non-small-cell lung carcinoma (NSCLC), African-Americans have lower survival rates than non-African-Americans. Whether this difference is due to innate characteristics of the disease in the two ethnicities or to disparities in health care is not known. We investigated whether the disparity in survival would persist when patients were treated with similar systemic therapies (i.e., in phase II and phase III Cancer and Leukemia Group B [CALGB] trials). METHODS: We assessed 504 consecutive patients (458 non African-American and 46 African-American) receiving systemic chemotherapy in CALGB studies for advanced NSCLC during the period from 1989 through 1998. Clinical and demographic characteristics, treatment received, and survival data were obtained from the CALGB database. Cox's proportional hazards model was used to assess the effect of race/ethnicity on survival after adjustment for other known prognostic factors. All statistical tests were two-sided. RESULTS: The unadjusted 1-year survival rate was 22% (95% confidence interval [CI] = 13% to 38%) for African-American patients and 30% (95% CI = 26% to 35%) for non-African American patients, a statistically significant difference (8%; 95% CI on the difference = 5% to 12%; P =.03). Multivariable adjustment for the effect of treatment arm, histology, and metastatic site at presentation did not alter the worse outcome for African-American patients. However, the effect of race/ethnicity disappeared after adjustment for performance status and weight loss. African-American patients were more likely than non-African-Americans to present with a poor performance status (83% versus 60%) and substantial weight loss (41% versus 27%) and to be unmarried (59% versus 28%), disabled (31% versus 15%), unemployed (17% versus 7%), and Medicaid recipients (30% versus 8%). CONCLUSIONS: The relationship that we observed between poor performance, weight loss, and socioeconomic status suggests that social circumstances lead to African Americans presenting with poorer prognostic features. PMID- 11854391 TI - Economic analysis of vinorelbine plus cisplatin versus paclitaxel plus carboplatin for advanced non-small-cell lung cancer. AB - BACKGROUND: It is increasingly important to have timely information about the economic impact of new cancer therapies in today's cost-conscious environment. Nearly 170 000 people are diagnosed with lung cancer annually in the United States. We performed an economic analysis alongside Southwest Oncology Group Trial S9509 to estimate the cost-effectiveness of cisplatin plus vinorelbine versus carboplatin plus paclitaxel for patients with advanced non-small-cell lung cancer. There were no statistically significant differences in survival or cancer related quality of life between the treatment arms. METHODS: Use of both protocol and nonprotocol lung cancer-related health care was tracked for 24 months from the initiation of therapy. To determine expenditures, nationally standardized costs were applied to each type of health care service used, and these were summed over time. Lifetime expenditures and 95% confidence intervals (CIs) for each arm of the trial were calculated with the use of a multivariate regression technique that accounts for censoring. Student's t tests were used to compare the difference in costs between the arms. All statistical tests were two-sided. RESULTS: Cancer-related health care costs over the period of observation averaged 40,292 dollars (95% CI = 36,226 dollars to 44,359 dollars) for patients in the cisplatin plus vinorelbine arm versus 48,940 dollars (95% CI = 44,674 dollars to 53,208 dollars) for patients in the carboplatin plus paclitaxel arm (P =.004), with a mean difference of 8648 dollars (95% CI = 2634 dollars to 14,662 dollars). Protocol chemotherapy drugs and medical procedures costs were statistically significantly higher in the paclitaxel arm (P =.0003 and P<.0001, respectively), whereas protocol chemotherapy delivery costs were statistically significantly higher in the vinorelbine arm (P<.0001). There was no difference between the arms in costs for blood products, supportive care medications, nonprotocol-related inpatient or outpatient care, and nonprotocol chemotherapy. CONCLUSIONS: Treatment with carboplatin plus paclitaxel is substantially and statistically significantly more expensive than treatment with cisplatin plus vinorelbine. The majority of the cost difference is due to the additional cost of the protocol chemotherapy (approximately 12,000 dollars). Notable differences in costs of downstream health care were not apparent. PMID- 11854392 TI - A population-based, case-control study of polymorphisms in carcinogen metabolizing genes, smoking, and pancreatic adenocarcinoma risk. AB - BACKGROUND: Cigarette smoking is associated with a twofold increased risk of pancreatic cancer. We conducted a population-based case-control study in six San Francisco Bay area counties from 1994 to 2001 to investigate associations between polymorphisms in genes for two carcinogen-metabolizing enzymes (cytochrome P450 1A1 [CYP1A1] and glutathione S-transferase [GST]), smoking, and adenocarcinoma of the exocrine pancreas. METHODS: We used polymerase chain reaction-based methods to analyze blood samples obtained from 309 case subjects and 964 control subjects to determine their genotypes for three CYP1A1 polymorphisms (m1, m2, and m4) and for homozygous deletions of two GST genes, GSTM1 and GSTT1. Control subjects were frequency matched to case subjects by age and sex. All statistical tests were two sided. RESULTS: None of the genetic polymorphisms themselves affected the risk of pancreatic cancer among Caucasian study participants. However, we observed an interaction between GSTT1-null genotype and cigarette smoking among Caucasians that was more prominent among women than among men. Relative to never smokers with the GSTT1-present genotype, the age-adjusted odds ratios (ORs) of pancreatic cancer for heavy smokers with the GSTT1-null genotype were 5.0 (95% confidence interval [CI] = 1.8 to 14.5) for women and 3.2 (95% CI = 1.3 to 8.1) for men; for heavy smokers with the GSTT1-present genotype they were 2.0 (95% CI = 1.0 to 4.0) for women and 2.1 (95% CI = 1.1 to 3.9) for men. ORs for pancreatic cancer among heavy smokers with both GSTT1-null and GSTM1-null genotypes were similar in magnitude to those among heavy smokers with the GSTT1-null genotype alone. There was no evidence of an interaction between CYP1A1 polymorphisms and smoking. CONCLUSIONS: The combination of heavy smoking and a deletion polymorphism in GSTT1 is associated with an increased risk of pancreatic cancer among Caucasians, with the associations possibly stronger in women than in men. PMID- 11854394 TI - Re: risk-reduction mastectomy: clinical issues and research needs. PMID- 11854395 TI - Re: sex, smoking, and cancer: a reappraisal. PMID- 11854396 TI - Re: sex, smoking, and cancer: a reappraisal. PMID- 11854398 TI - Post-cytochrome C protection from apoptosis conferred by a MAPK pathway in Xenopus egg extracts. AB - In response to many different apoptotic stimuli, cytochrome c is released from the intermembrane space of the mitochondria into the cytoplasm, where it serves as a cofactor in the activation of procaspase 9. Inhibition of this process can occur either by preventing cytochrome c release or by blocking caspase activation or activity. Experiments involving in vitro reconstitution of apoptosis in cell free extracts of Xenopus laevis eggs have suggested that extracts arrested in interphase are susceptible to an endogenous apoptotic program leading to caspase activation, whereas extracts arrested in meiotic metaphase are not. We report here that Mos/MEK/MAPK pathways active in M phase-arrested eggs are responsible for rendering them refractory to apoptosis. Interestingly, M phase-arrested extracts are competent to release cytochrome c, yet still do not activate caspases. Concomitantly, we have also demonstrated that recombinant Mos, MEK, and ERK are sufficient to block cytochrome c-dependent caspase activation in purified Xenopus cytosol, which lacks both transcription and translation. These data indicate that the MAP kinase pathway can target and inhibit post-cytochrome c release apoptotic events in the absence of new mRNA/protein synthesis and that this biochemical pathway is responsible for the apoptotic inhibition observed in meiotic X. laevis egg extracts. PMID- 11854399 TI - Dynamics of cytoskeletal proteins during Fcgamma receptor-mediated phagocytosis in macrophages. AB - Particle ingestion by phagocytosis results from sequential rearrangements of the actin cytoskeleton and overlying membrane. To assemble a chronology of molecular events during phagosome formation and to examine the contributions of phosphoinositide 3-kinase (PI 3-kinase) to these dynamics, a method was developed for synchronizing Fcgamma receptor-mediated phagocytosis by murine macrophages. Erythrocytes opsonized with complement component C3bi were bound to macrophages at 37degrees C, a condition that does not favor particle phagocytosis. Addition of soluble anti-erythrocyte IgG resulted in rapid opsonization of the bound erythrocytes, followed by their immediate internalization via phagocytosis. Cellular content of F-actin, as measured by binding of rhodamine-phalloidin, increased transiently during phagocytosis, and this increase was not diminished by inhibitors of PI 3-kinase. Immunofluorescence localization of myosins in macrophages fixed at various times during phagocytosis indicated that myosins II and IXb were concentrated in early phagosomes, myosin IC increased later, and myosin V appeared after phagosome closure. Other cytoskeletal proteins showed similar variations in the timing of their appearance in phagosomes. The PI 3 kinase inhibitor wortmannin did not change the dynamics of PI 3-kinase or ezrin localization but prevented the loss of PAK1 from phagosomes. These results suggest that PI 3-kinase deactivates PAK1, and that this may be needed for phagosome closure. PMID- 11854401 TI - Nuclear pore complex is able to transport macromolecules with diameters of about 39 nm. AB - Bidirectional transport of macromolecules between the nucleus and the cytoplasm occurs through the nuclear pore complexes (NPCs) by a signal-mediated mechanism that is directed by targeting signals (NLSs) residing on the transported molecules or "cargoes." Nuclear transport starts after interaction of the targeting signal with soluble cellular receptors. After the formation of the cargo-receptor complex in the cytosol, this complex crosses the NPC. Herein, we use gold particles of various sizes coated with cargo-receptor complexes to determine precisely how large macromolecules crossing the NPC by the signal mediated transport mechanism could be. We found that cargo-receptor-gold complexes with diameter close to 39 nm could be translocated by the NPC. This implies that macromolecules much larger than the assumed functional NPC diameter of 26 nm can be transported into the karyoplasm. The physiological relevance of this finding was supported by the observation that intact nucleocapsids of human hepatitis B virus with diameters of 32 and 36 nm are able to cross the nuclear pore without disassembly. PMID- 11854400 TI - The eukaryotic two-component histidine kinase Sln1p regulates OCH1 via the transcription factor, Skn7p. AB - The yeast "two-component" osmotic stress phosphorelay consists of the histidine kinase, Sln1p, the phosphorelay intermediate, Ypd1p and two response regulators, Ssk1p and Skn7p, whose activities are regulated by phosphorylation of a conserved aspartyl residue in the receiver domain. Dephospho-Ssk1p leads to activation of the hyper-osmotic response (HOG) pathway, whereas phospho-Skn7p presumably leads to activation of hypo-osmotic response genes. The multifunctional Skn7 protein is important in oxidative as well as osmotic stress; however, the Skn7p receiver domain aspartate that is the phosphoacceptor in the SLN1 pathway is dispensable for oxidative stress. Like many well-characterized bacterial response regulators, Skn7p is a transcription factor. In this report we investigate the role of Skn7p in osmotic response gene activation. Our studies reveal that the Skn7p HSF-like DNA binding domain interacts with a cis-acting element identified upstream of OCH1 that is distinct from the previously defined HSE-like Skn7p binding site. Our data support a model in which Skn7p receiver domain phosphorylation affects transcriptional activation rather than DNA binding to this class of DNA binding site. PMID- 11854403 TI - RNAi in Dictyostelium: the role of RNA-directed RNA polymerases and double stranded RNase. AB - We show that in Dictyostelium discoideum an endogenous gene as well as a transgene can be silenced by introduction of a gene construct that is transcribed into a hairpin RNA. Gene silencing was accompanied by the appearance of sequence specific RNA about 23mers and seemed to have a limited capacity. The three Dictyostelium homologues of the RNA-directed RNA polymerase (RrpA, RrpB, and DosA) all contain an N-terminal helicase domain homologous to the one in the dicer nuclease, suggesting exon shuffling between RNA-directed RNA polymerase and the dicer homologue. Only the knock-out of rrpA resulted in a loss of the hairpin RNA effect and simultaneously in a loss of detectable about 23mers. However, about 23mers were still generated by the Dictyostelium dsRNase in vitro with extracts from rrpA(-), rrpB(-), and DosA(-) cells. Both RrpA and a target gene were required for production of detectable amounts of about 23mers, suggesting that target sequences are involved in about 23mer amplification. PMID- 11854402 TI - Essential role of MCM proteins in premeiotic DNA replication. AB - A critical event in eukaryotic DNA replication involves association of minichromosome maintenance (MCM2-7) proteins with origins, to form prereplicative complexes (pre-RCs) that are competent for initiation. The ability of mutants defective in MCM2-7 function to complete meiosis had suggested that pre-RC components could be irrelevant to premeiotic S phase. We show here that MCM2-7 proteins bind to chromatin in fission yeast cells preparing for meiosis and during premeiotic S phase in a manner suggesting they in fact are required for DNA replication in the meiotic cycle. This is confirmed by analysis of a degron mcm4 mutant, which cannot carry out premeiotic DNA replication. Later in meiosis, Mcm4 chromatin association is blocked between meiotic nuclear divisions, presumably accounting for the absence of a second round of DNA replication. Together, these results emphasize similarity between replication mechanisms in mitotic and meiotic cell cycles. PMID- 11854405 TI - Functional characterization and localization of the Aspergillus nidulans formin SEPA. AB - Formins are a family of multidomain scaffold proteins involved in actin-dependent morphogenetic events. In Aspergillus nidulans, the formin SEPA participates in two actin-mediated processes, septum formation and polarized growth. In this study, we use a new null mutant to demonstrate that SEPA is required for the formation of actin rings at septation sites. In addition, we find that a functional SEPA::GFP fusion protein localizes simultaneously to septation sites and hyphal tips, and that SEPA colocalizes with actin at each site. Using live imaging, we show that SEPA localization at septation sites and hyphal tips is dynamic. Notably, at septation sites, SEPA forms a ring that constricts as the septum is deposited. Moreover, we demonstrate that actin filaments are required to maintain the proper localization pattern of SEPA, and that the amino-terminal half of SEPA is sufficient for localization at septation sites and hyphal tips. In contrast, only localization at septation sites is affected by loss of the sepH gene product. We propose that specific morphological cues activate common molecular pathways to direct SEPA localization to the appropriate morphogenetic site. PMID- 11854404 TI - Activation of p42 mitogen-activated protein kinase (MAPK), but not c-Jun NH(2) terminal kinase, induces phosphorylation and stabilization of MAPK phosphatase XCL100 in Xenopus oocytes. AB - Dual-specificity protein phosphatases are implicated in the direct down regulation of mitogen-activated protein kinase (MAPK) activity in vivo. Accumulating evidence suggests that these phosphatases are components of negative feedback loops that restore MAPK activity to low levels after diverse physiological responses. Limited information exists, however, regarding their posttranscriptional regulation. We cloned two Xenopus homologs of the mammalian dual-specificity MAPK phosphatases MKP-1/CL100 and found that overexpression of XCL100 in G2-arrested oocytes delayed or prevented progesterone-induced meiotic maturation. Epitope-tagged XCL100 was phosphorylated on serine during G2 phase, and on serine and threonine in a p42 MAPK-dependent manner during M phase. Threonine phosphorylation mapped to a single residue, threonine 168. Phosphorylation of XCL100 had no measurable effect on its ability to dephosphorylate p42 MAPK. Similarly, mutation of threonine 168 to either valine or glutamate did not significantly alter the binding affinity of a catalytically inactive XCL100 protein for active p42 MAPK in vivo. XCL100 was a labile protein in G2-arrested and progesterone-stimulated oocytes; surprisingly, its degradation rate was increased more than twofold after exposure to hyperosmolar sorbitol. In sorbitol-treated oocytes expressing a conditionally active DeltaRaf-DD:ER chimera, activation of the p42 MAPK cascade led to phosphorylation of XCL100 and a pronounced decrease in the rate of its degradation. Our results provide mechanistic insight into the regulation of a dual-specificity MAPK phosphatase during meiotic maturation and the adaptation to cellular stress. PMID- 11854407 TI - Deletion mutants in COP9/signalosome subunits in fission yeast Schizosaccharomyces pombe display distinct phenotypes. AB - The COP9/signalosome complex is highly conserved in evolution and possesses significant structural similarity to the 19S regulatory lid complex of the proteasome. It also shares limited similarity to the translation initiation factor eIF3. The signalosome interacts with multiple cullins in mammalian cells. In the fission yeast Schizosaccharomyces pombe, the Csn1 subunit is required for the removal of covalently attached Nedd8 from Pcu1, one of three S. pombe cullins. It remains unclear whether this activity is required for all the functions ascribed to the signalosome. We previously identified Csn1 and Csn2 as signalosome subunits in S. pombe. csn1 and csn2 null mutants are DNA damage sensitive and exhibit slow DNA replication. Two further putative subunits, Csn4 and Csn5, were identified from the S. pombe genome database. Herein, we characterize null mutations of csn4 and csn5 and demonstrate that both genes are required for removal of Nedd8 from the S. pombe cullin Pcu1 and that their protein products associate with Csn1 and Csn2. However, neither csn4 nor csn5 null mutants share the csn1 and csn2 mutant phenotypes. Our data suggest that the subunits of the signalosome cannot be considered as a distinct functional unit and imply that different subunits of the signalosome mediate distinct functions. PMID- 11854406 TI - Fission yeast Rad26 is a regulatory subunit of the Rad3 checkpoint kinase. AB - Fission yeast Rad3 is a member of a family of phosphoinositide 3-kinase -related kinases required for the maintenance of genomic stability in all eukaryotic cells. In fission yeast, Rad3 regulates the cell cycle arrest and recovery activities associated with the G2/M checkpoint. We have developed an assay that directly measures Rad3 kinase activity in cells expressing physiological levels of the protein. Using the assay, we demonstrate directly that Rad3 kinase activity is stimulated by checkpoint signals. Of the five other G2/M checkpoint proteins (Hus1, Rad1, Rad9, Rad17, and Rad26), only Rad26 was required for Rad3 kinase activity. Because Rad26 has previously been shown to interact constitutively with Rad3, our results demonstrate that Rad26 is a regulatory subunit, and Rad3 is the catalytic subunit, of the Rad3/Rad26 kinase complex. Analysis of Rad26/Rad3 kinase activation in rad26.T12, a mutant that is proficient for cell cycle arrest, but defective in recovery, suggests that these two responses to checkpoint signals require quantitatively different levels of kinase activity from the Rad3/Rad26 complex. PMID- 11854408 TI - Pag1p, a novel protein associated with protein kinase Cbk1p, is required for cell morphogenesis and proliferation in Saccharomyces cerevisiae. AB - Protein kinases in the Cot-1/Orb6/Ndr/Warts family are important regulators of cell morphogenesis and proliferation. Cbk1p, a member of this family in Saccharomyces cerevisiae, has previously been shown to be required for normal morphogenesis in vegetatively growing cells and in haploid cells responding to mating pheromone. A mutant of PAG1, a novel gene in S. cerevisiae, displayed defects similar to those of cbk1 mutants. pag1 and cbk1 mutants share a common set of suppressors, including the disruption of SSD1, a gene encoding an RNA binding protein, and the overexpression of Sim1p, an extracellular protein. These genetic results suggest that PAG1 and CBK1 act in the same pathway. Furthermore, we found that Pag1p and Cbk1p localize to the same polarized peripheral sites and that they coimmunoprecipitate with each other. Pag1p is a conserved protein. The homologs of Pag1p in other organisms are likely to form complexes with the Cbk1p related kinases and function with those kinases in the same biological processes. PMID- 11854409 TI - The multiprotein exocyst complex is essential for cell separation in Schizosaccharomyces pombe. AB - Schizosaccharomyces pombe cells divide by medial fission through the use of an actomyosin-based contractile ring. A mulitlayered division septum is assembled in concert with ring constriction. Finally, cleavage of the inner layer of the division septum results in the liberation of daughter cells. Although numerous studies have focused on actomyosin ring and division septum assembly, little information is available on the mechanism of cell separation. Here we describe a mutant, sec8-1, that is defective in cell separation but not in other aspects of cytokinesis. sec8-1 mutants accumulate about 100-nm vesicles and have reduced secretion of acid phosphatase, suggesting that they are defective in exocytosis. Sec8p is a component of the exocyst complex. Using biochemical methods, we show that Sec8p physically interacts with other members of the exocyst complex, including Sec6p, Sec10p, and Exo70p. These exocyst proteins localize to regions of active exocytosis-at the growing ends of interphase cells and in the medial region of cells undergoing cytokinesis-in an F-actin-dependent and exocytosis independent manner. Analysis of a number of mutations in various exocyst components has established that these components are essential for cell viability. Interestingly, all exocyst mutants analyzed appear to be able to elongate and to assemble division septa but are defective for cell separation. We therefore propose that the fission yeast exocyst is involved in targeting of enzymes responsible for septum cleavage. We further propose that cell elongation and division septum assembly can continue with minimal levels of exocyst function. PMID- 11854410 TI - Secretion and assembly of zona pellucida glycoproteins by growing mouse oocytes microinjected with epitope-tagged cDNAs for mZP2 and mZP3. AB - The zona pellucida (ZP) is a highly organized extracellular coat that surrounds all mammalian eggs. The mouse egg ZP is composed of three glycoproteins, called mZP1-3, that are synthesized, secreted, and assembled into a ZP exclusively by growing oocytes. Here, we microinjected epitope-tagged (Myc and Flag) cDNAs for mZP2 and mZP3 into the germinal vesicle (nucleus) of growing oocytes isolated from juvenile mice. Specific antibodies and laser scanning confocal microscopy were used to follow nascent, recombinant ZP glycoproteins in both permeabilized and nonpermeabilized oocytes. When such cDNAs were injected, epitope-tagged mZP2 (Myc-mZP2) and mZP3 (Flag-mZP3) were synthesized, packaged into large intracellular vesicles, and secreted by the vast majority of oocytes. Secreted glycoproteins were incorporated into only the innermost layer of the thickening ZP, and the amount of nascent glycoprotein in this region increased with increasing time of oocyte culture. Consistent with prior observations, the putative transmembrane domain at the C terminus of mZP2 and mZP3 was missing from nascent glycoprotein incorporated into the ZP. When the consensus furin cleavage site near the C terminus of mZP3 was mutated, such that it should not be cleaved by furin, secretion and assembly of mZP3 was reduced. On the other hand, mZP3 incorporated into the ZP lacked the transmembrane domain downstream of the mutated furin cleavage site, suggesting that some other protease(s) excised the domain. These results strongly suggest that nascent mZP2 and mZP3 are incorporated into only the innermost layer of the ZP and that excision of the C terminal region of the glycoproteins is required for assembly into the oocyte ZP. PMID- 11854411 TI - The yeast synaptojanin-like proteins control the cellular distribution of phosphatidylinositol (4,5)-bisphosphate. AB - Phosphoinositides (PI) are synthesized and turned over by specific kinases, phosphatases, and lipases that ensure the proper localization of discrete PI isoforms at distinct membranes. We analyzed the role of the yeast synaptojanin like proteins using a strain that expressed only a temperature-conditional allele of SJL2. Our analysis demonstrated that inactivation of the yeast synaptojanins leads to increased cellular levels of phosphatidylinositol (3,5)-bisphosphate and phosphatidylinositol (4,5)-bisphosphate (PtdIns(4,5)P(2)), accompanied by defects in actin organization, endocytosis, and clathrin-mediated sorting between the Golgi and endosomes. The phenotypes observed in synaptojanin-deficient cells correlated with accumulation of PtdIns(4,5)P(2), because these effects were rescued by mutations in MSS4 or a mutant form of Sjl2p that harbors only PI 5 phosphatase activity. We utilized green fluorescent protein-pleckstrin homology domain chimeras (termed FLAREs for fluorescent lipid-associated reporters) with distinct PI-binding specificities to visualize pools of PtdIns(4,5)P(2) and phosphatidylinositol 4-phosphate in yeast. PtdIns(4,5)P(2) localized to the plasma membrane in a manner dependent on Mss4p activity. On inactivation of the yeast synaptojanins, PtdIns(4,5)P(2) accumulated in intracellular compartments, as well as the cell surface. In contrast, phosphatidylinositol 4-phosphate generated by Pik1p localized in intracellular compartments. Taken together, our results demonstrate that the yeast synaptojanins control the localization of PtdIns(4,5)P(2) in vivo and provide further evidence for the compartmentalization of different PI species. PMID- 11854412 TI - The Drosophila nuclear lamina protein YA binds to DNA and histone H2B with four domains. AB - Dramatic changes occur in nuclear organization and function during the critical developmental transition from meiosis to mitosis. The Drosophila nuclear lamina protein YA binds to chromatin and is uniquely required for this transition. In this study, we dissected YA's binding to chromatin. We found that YA can bind to chromatin directly and specifically. It binds to DNA but not RNA, with a preference for double-stranded DNA (linear or supercoiled) over single-stranded DNA. It also binds to histone H2B. YA's binding to DNA and histone H2B is mediated by four domains distributed along the length of the YA molecule. A model for YA function at the end of Drosophila female meiosis is proposed. PMID- 11854413 TI - Macromolecular uptake is a spontaneous event during mitosis in cultured fibroblasts: implications for vector-dependent plasmid transfection. AB - The process through which macromolecules penetrate the plasma membrane of mammalian cells remains poorly defined. We have examined whether natural cellular events modulate the capacity of cells to take up agents applied extraneously. Herein, we report that during mitosis and in a cell type-independent manner, cells exhibit a natural ability to absorb agents present in the extracellular environment up to 150 kDa as assessed using fluorescein isothiocyanate-dextrans. This event is exclusive to the mitotic period and not observed during G0, G1, S, or G2 phase. During mitosis, starting in advanced prophase, oligonucleotides, active enzymes, and polypeptides are efficiently taken into mitotic cells. This uptake of macromolecules during mitosis still takes place in the presence of cytochalasin D or nocodazole, showing no requirement for intact microtubules or actin filaments in this process. However, cell rounding up, which still takes place in the presence of either of these drugs in mitotic cells, appears to be a key event in this process. Indeed, limited trypsinization of adherent cells mimics both the cell retraction and macromolecule uptake observed as cells enter mitosis. A plasmid DNA encoding green fluorescent protein (3.3Mda) coated with an 18 amino acid peptide is efficiently expressed when applied onto synchronized G2/M fibroblasts, whereas little or no expression is observed when the coated plasmid is applied onto asynchronous cell cultures. This shows that such coating peptides are only efficient for their encapsulating and protective effect on the plasmid DNA to be "vectorized" rather than acting as true vectors. PMID- 11854414 TI - Interactions of elongation factor 1alpha with F-actin and beta-actin mRNA: implications for anchoring mRNA in cell protrusions. AB - The targeting of mRNA and local protein synthesis is important for the generation and maintenance of cell polarity. As part of the translational machinery as well as an actin/microtubule-binding protein, elongation factor 1alpha (EF1alpha) is a candidate linker between the protein translation apparatus and the cytoskeleton. We demonstrate in this work that EF1alpha colocalizes with beta-actin mRNA and F actin in protrusions of chicken embryo fibroblasts and binds directly to F-actin and beta-actin mRNA simultaneously in vitro in actin cosedimentation and enzyme linked immunosorbent assays. To investigate the role of EF1alpha in mRNA targeting, we mapped the two actin-binding sites on EF1alpha at high resolution and defined one site at the N-terminal 49 residues of domain I and the other at the C-terminal 54 residues of domain III. In vitro actin-binding assays and localization in vivo of recombinant full-length EF1alpha and its various truncates demonstrated that the C terminus of domain III was the dominant actin binding site both in vitro and in vivo. We propose that the EF1alpha-F-actin complex is the scaffold that is important for beta-actin mRNA anchoring. Disruption of this complex would lead to delocalization of the mRNA. This hypothesis was tested by using two dominant negative polypeptides: the actin binding domain III of EF1alpha and the EF1alpha-binding site of yeast Bni1p, a protein that inhibits EF1alpha binding to F-actin and also is required for yeast mRNA localization. We demonstrate that either domain III of EF1alpha or the EF1alpha-binding site of Bni1p inhibits EF1alpha binding to beta-actin mRNA in vitro and causes delocalization of beta-actin mRNA in chicken embryo fibroblasts. Taken together, these results implicate EF1alpha in the anchoring of beta-actin mRNA to the protrusion in crawling cells. PMID- 11854415 TI - Daughter cell assembly in the protozoan parasite Toxoplasma gondii. AB - The phylum Apicomplexa includes thousands of species of obligate intracellular parasites, many of which are significant human and/or animal pathogens. Parasites in this phylum replicate by assembling daughters within the mother, using a cytoskeletal and membranous scaffolding termed the inner membrane complex. Most apicomplexan parasites, including Plasmodium sp. (which cause malaria), package many daughters within a single mother during mitosis, whereas Toxoplasma gondii typically packages only two. The comparatively simple pattern of T. gondii cell division, combined with its molecular genetic and cell biological accessibility, makes this an ideal system to study parasite cell division. A recombinant fusion between the fluorescent protein reporter YFP and the inner membrane complex protein IMC1 has been exploited to examine daughter scaffold formation in T. gondii. Time-lapse video microscopy permits the entire cell cycle of these parasites to be visualized in vivo. In addition to replication via endodyogeny (packaging two parasites at a time), T. gondii is also capable of forming multiple daughters, suggesting fundamental similarities between cell division in T. gondii and other apicomplexan parasites. PMID- 11854416 TI - Drosophila minichromosome maintenance 6 is required for chorion gene amplification and genomic replication. AB - Duplication of the eukaryotic genome initiates from multiple origins of DNA replication whose activity is coordinated with the cell cycle. We have been studying the origins of DNA replication that control amplification of eggshell (chorion) genes during Drosophila oogenesis. Mutation of genes required for amplification results in a thin eggshell phenotype, allowing a genetic dissection of origin regulation. Herein, we show that one mutation corresponds to a subunit of the minichromosome maintenance (MCM) complex of proteins, MCM6. The binding of the MCM complex to origins in G1 as part of a prereplicative complex is critical for the cell cycle regulation of origin licensing. We find that MCM6 associates with other MCM subunits during amplification. These results suggest that chorion origins are bound by an amplification complex that contains MCM proteins and therefore resembles the prereplicative complex. Lethal alleles of MCM6 reveal it is essential for mitotic cycles and endocycles, and suggest that its function is mediated by ATP. We discuss the implications of these findings for the role of MCMs in the coordination of DNA replication during the cell cycle. PMID- 11854417 TI - Golgi vesicle proteins are linked to the assembly of an actin complex defined by mAbp1. AB - Recent studies indicate that regulation of the actin cytoskeleton is important for protein trafficking, but its precise role is unclear. We have characterized the ARF1-dependent assembly of actin on the Golgi apparatus. Actin recruitment involves Cdc42/Rac and requires the activation of the Arp2/3 complex. Although the actin-binding proteins mAbp1 (SH3p7) and drebrin share sequence homology, they are differentially segregated into two distinct ARF-dependent actin complexes. The binding of Cdc42 and mAbp1, which localize to the Golgi apparatus, but not drebrin, is blocked by occupation of the p23 cargo-protein-binding site on coatomer. Exogenously expressed mAbp1 is mislocalized and inhibits Golgi transport in whole cells. The ability of ARF, vesicle-coat proteins, and cargo to direct the assembly of cytoskeletal structures helps explain how only a handful of vesicle types can mediate the numerous trafficking steps in the cell. PMID- 11854418 TI - Mutation of YCS4, a budding yeast condensin subunit, affects mitotic and nonmitotic chromosome behavior. AB - The budding yeast YCS4 gene encodes a conserved regulatory subunit of the condensin complex. We isolated an allele of this gene in a screen for mutants defective in sister chromatid separation or segregation. The phenotype of the ycs4-1 mutant is similar to topoisomerase II mutants and distinct from the esp1-1 mutant: the topological resolution of sister chromatids is compromised in ycs4-1 despite normal removal of cohesins from mitotic chromosomes. Consistent with a role in sister separation, YCS4 function is required to localize DNA topoisomerase I and II to chromosomes. Unlike its homologs in Xenopus and fission yeast, Ycs4p is associated with chromatin throughout the cell cycle; the only change in localization occurs during anaphase when the protein is enriched at the nucleolus. This relocalization may reveal the specific challenge that segregation of the transcriptionally hyperactive, repetitive array of rDNA genes can present during mitosis. Indeed, segregation of the nucleolus is abnormal in ycs4-1 at the nonpermissive temperature. Interrepeat recombination in the rDNA array is specifically elevated in ycs4-1 at the permissive temperature, suggesting that the Ycs4p plays a role at the array aside from its segregation. Furthermore, ycs4 1 is defective in silencing at the mating type loci at the permissive temperature. Taken together, our data suggest that there are mitotic as well as nonmitotic chromosomal abnormalities associated with loss of condensin function in budding yeast. PMID- 11854419 TI - CSN1 N-terminal-dependent activity is required for Arabidopsis development but not for Rub1/Nedd8 deconjugation of cullins: a structure-function study of CSN1 subunit of COP9 signalosome. AB - The COP9 signalosome (CSN) is a multifunctional protein complex essential for arabidopsis development. One of its functions is to promote Rub1/Nedd8 deconjugation from the cullin subunit of the Skp1-cullin-F-box ubiquitin ligase. Little is known about the specific role of its eight subunits in deneddylation or any of the physiological functions of CSN. In the absence of CSN1 (the fus6 mutant), arabidopsis CSN complex cannot assemble, which destabilizes multiple CSN subunits and contributes, together with the loss of CSN1, to the phenotype of fus6. To distinguish CSN1-specific functions, we attempted to rescue the complex formation with deletion or point-mutation forms of CSN1 expressed as transgenes in fus6. We show that the central domain of CSN1 is critical for complex assembly, whereas the C-terminal domain has a supporting role. By expressing the C231 fragment, which contains the structural information but lacks the presumed functional domain located at the N terminus, we have rescued the complex formation and restored the Rub1/Nedd8 deconjugation activity on cullins (fus6/C231). Nonetheless, fus6/C231 exhibits pleiotropic phenotype, including photomorphogenic defects and growth arrest at seedling stage. We conclude that CSN1 N-terminal domain is not required for the Rub1/Nedd8 deconjugation activity of cullins, but contributes to a significant aspect of CSN functions that are essential for plant development. PMID- 11854420 TI - Dictyostelium LvsB mutants model the lysosomal defects associated with Chediak Higashi syndrome. AB - Chediak-Higashi syndrome is a genetic disorder caused by mutations in a gene encoding a protein named LYST in humans ("lysosomal trafficking regulator") or Beige in mice. A prominent feature of this disease is the accumulation of enlarged lysosome-related granules in a variety of cells. The genome of Dictyostelium discoideum contains six genes encoding proteins that are related to LYST/Beige in amino acid sequence, and disruption of one of these genes, lvsA (large volume sphere), results in profound defects in cytokinesis. To better understand the function of this family of proteins in membrane trafficking, we have analyzed mutants disrupted in lvsA, lvsB, lvsC, lvsD, lvsE, and lvsF. Of all these, only lvsA and lvsB mutants displayed interesting phenotypes in our assays. lvsA-null cells exhibited defects in phagocytosis and contained abnormal looking contractile vacuole membranes. Loss of LvsB, the Dictyostelium protein most similar to LYST/Beige, resulted in the formation of enlarged vesicles that by multiple criteria appeared to be acidic lysosomes. The rates of endocytosis, phagocytosis, and fluid phase exocytosis were normal in lvsB-null cells. Also, the rates of processing and the efficiency of targeting of lysosomal alpha mannosidase were normal, although lvsB mutants inefficiently retained alpha mannosidase, as well as two other lysosomal cysteine proteinases. Finally, results of pulse-chase experiments indicated that an increase in fusion rates accounted for the enlarged lysosomes in lvsB-null cells, suggesting that LvsB acts as a negative regulator of fusion. Our results support the notion that LvsB/LYST/Beige function in a similar manner to regulate lysosome biogenesis. PMID- 11854421 TI - Identification and characterization of ART-27, a novel coactivator for the androgen receptor N terminus. AB - The androgen receptor (AR) is a ligand-regulated transcription factor that stimulates cell growth and differentiation in androgen-responsive tissues. The AR N terminus contains two activation functions (AF-1a and AF-1b) that are necessary for maximal transcriptional enhancement by the receptor; however, the mechanisms and components regulating AR transcriptional activation are not fully understood. We sought to identify novel factors that interact with the AR N terminus from an androgen-stimulated human prostate cancer cell library using a yeast two-hybrid approach designed to identify proteins that interact with transcriptional activation domains. A 157-amino acid protein termed ART-27 was cloned and shown to interact predominantly with the AR(153-336), containing AF-1a and a part of AF 1b, localize to the nucleus and increase the transcriptional activity of AR when overexpressed in cultured mammalian cells. ART-27 also enhanced the transcriptional activation by AR(153-336) fused to the LexA DNA-binding domain but not other AR N-terminal subdomains, suggesting that ART-27 exerts its effect via an interaction with a defined region of the AR N terminus. ART-27 interacts with AR in nuclear extracts from LNCaP cells in a ligand-independent manner. Interestingly, velocity gradient sedimentation of HeLa nuclear extracts suggests that native ART-27 is part of a multiprotein complex. ART-27 is expressed in a variety of human tissues, including sites of androgen action such as prostate and skeletal muscle, and is conserved throughout evolution. Thus, ART-27 is a novel cofactor that interacts with the AR N terminus and plays a role in facilitating receptor-induced transcriptional activation. PMID- 11854423 TI - Drosophila stathmin: a microtubule-destabilizing factor involved in nervous system formation. AB - Stathmin is a ubiquitous regulatory phosphoprotein, the generic element of a family of neural phosphoproteins in vertebrates that possess the capacity to bind tubulin and interfere with microtubule dynamics. Although stathmin and the other proteins of the family have been associated with numerous cell regulations, their biological roles remain elusive, as in particular inactivation of the stathmin gene in the mouse resulted in no clear deleterious phenotype. We identified stathmin phosphoproteins in Drosophila, encoded by a unique gene sharing the intron/exon structure of the vertebrate stathmin and stathmin family genes. They interfere with microtubule assembly in vitro, and in vivo when expressed in HeLa cells. Drosophila stathmin expression is regulated during embryogenesis: it is high in the migrating germ cells and in the central and peripheral nervous systems, a pattern resembling that of mammalian stathmin. Furthermore, RNA interference inactivation of Drosophila stathmin expression resulted in germ cell migration arrest at stage 14. It also induced important anomalies in nervous system development, such as loss of commissures and longitudinal connectives in the ventral cord, or abnormal chordotonal neuron organization. In conclusion, a single Drosophila gene encodes phosphoproteins homologous to the entire vertebrate stathmin family. We demonstrate for the first time their direct involvement in major biological processes such as development of the reproductive and nervous systems. PMID- 11854422 TI - Pitx2a expression alters actin-myosin cytoskeleton and migration of HeLa cells through Rho GTPase signaling. AB - We ectopically expressed the transcription factor Pitx2a, one of the Pitx2 isoforms, in HeLa cells by using a tetracycline-inducible expression system and examined whether Pitx2a was capable of modulating Rho GTPase signaling and altering the cell's cytoskeleton. Ectopic expression of Pitx2a induced actin myosin reorganization, leading to increased cell spreading, suppression of cell migration, and the strengthening of cell-cell adhesion, marked by the accumulation and localization of beta-catenin and N-cadherin to the sites of cell cell contacts. Moreover, Pitx2a expression resulted in activation of the Rho GTPases Rac1 and RhoA, and the dominant negative Rac1 mutant N17Rac1 inhibited cell spreading and disrupted localization of beta-catenin to the sites of cell cell contacts. Both reorganization of actin-myosin and cell spreading require phosphatidylinositol 3-kinase activity, which is also necessary for activation of the Rho GTPase proteins. Pitx2a induced the expression of Trio, a guanine nucleotide exchange factor for Rac1 and RhoA, which preceded cell spreading, and the expression of Trio protein was down-regulated after the changes in cell spreading and cell morphology were initiated. In addition, Pitx2a also induces cell cycle arrest at G0/G1, most likely due to the accumulation of the tumor suppressor proteins p53 and p21. Our data indicate that the transcriptional activities initiated in the nucleus by Pitx2a result in profound changes in HeLa cell morphology, migration, and proliferation. PMID- 11854424 TI - Peroxisomes are formed from complex membrane structures in PEX6-deficient CHO cells upon genetic complementation. AB - Pex6p belongs to the AAA family of ATPases. Its CHO mutant, ZP92, lacks normal peroxisomes but contains peroxisomal membrane remnants, so called peroxisomal ghosts, which are detected with anti-70-kDa peroxisomal membrane protein (PMP70) antibody. No peroxisomal matrix proteins were detected inside the ghosts, but exogenously expressed green fluorescent protein (GFP) fused to peroxisome targeting signal-1 (PTS-1) accumulated in the areas adjacent to the ghosts. Electron microscopic examination revealed that PMP70-positive ghosts in ZP92 were complex membrane structures, rather than peroxisomes with reduced matrix protein import ability. In a typical case, a set of one central spherical body and two layers of double-membraned loops were observed, with endoplasmic reticulum present alongside the outer loop. In the early stage of complementation by PEX6 cDNA, catalase and acyl-CoA oxidase accumulated in the lumen of the double membraned loops. Biochemical analysis revealed that almost all the peroxisomal ghosts were converted into peroxisomes upon complementation. Our results indicate that 1) Peroxisomal ghosts are complex membrane structures; and 2) The complex membrane structures become import competent and are converted into peroxisomes upon complementation with PEX6. PMID- 11854426 TI - New determinants of receptor-effector coupling: trafficking and compartmentation in membrane microdomains. PMID- 11854425 TI - Multiple active states and oligomerization of CCR5 revealed by functional properties of monoclonal antibodies. AB - CC-chemokine receptor 5 (CCR5) is the principal coreceptor for macrophage-tropic strains of human immunodeficiency virus type 1 (HIV-1). We have generated a set of anti-CCR5 monoclonal antibodies and characterized them in terms of epitope recognition, competition with chemokine binding, receptor activation and trafficking, and coreceptor activity. MC-4, MC-5, and MC-7 mapped to the amino terminal domain, MC-1 to the second extracellular loop, and MC-6 to a conformational epitope covering multiple extracellular domains. MC-1 and MC-6 inhibited regulated on activation normal T cell expressed and secreted (RANTES), macrophage inflammatory polypeptide-1beta, and Env binding, whereas MC-5 inhibited macrophage inflammatory polypeptide-1beta and Env but not RANTES binding. MC-6 induced signaling in different functional assays, suggesting that this monoclonal antibody stabilizes an active conformation of CCR5. Flow cytometry and real-time confocal microscopy showed that MC-1 promoted strong CCR5 endocytosis. MC-1 but not its monovalent isoforms induced an increase in the transfer of energy between CCR5 molecules. Also, its monovalent isoforms bound efficiently, but did not internalize the receptor. In contrast, MC-4 did not prevent RANTES binding or subsequent signaling, but inhibited its ability to promote CCR5 internalization. These results suggest the existence of multiple active conformations of CCR5 and indicate that CCR5 oligomers are involved in an internalization process that is distinct from that induced by the receptor's agonists. PMID- 11854427 TI - Desensitization of cannabinoid-mediated presynaptic inhibition of neurotransmission between rat hippocampal neurons in culture. AB - Prolonged exposure to cannabinoids results in tolerance in vivo and desensitization of cannabinoid receptors in vitro. We show here that cannabinoid induced presynaptic inhibition of glutamatergic neurotransmission desensitized after prolonged exposure to the cannabinoid receptor agonist (R)-(+)-[2,3-dihydro 5-methyl-3-[(4-morpholinyl)methyl]pyrrolo-[1,2,3-de]-1,4-benzoxazin-6-yl](1 napthalenyl)methanone monomethanesulfonate (Win55,212-2). Synaptic activity between hippocampal neurons in culture was determined from network-driven increases in intracellular Ca(2+) concentration ([Ca(2+)](i) spikes) and excitatory postsynaptic currents. Win55,212-2-induced (100 nM) inhibition partially desensitized after 2 h and completely desensitized after 18- to 24-h exposure. The desensitization could be overcome by higher concentrations of agonist as indicated by a parallel rightward shift of the concentration response curve from an EC(50) of 2.7 +/- 0.3 nM to 320 +/- 147 nM for inhibition of [Ca(2+)](i) spiking and from 43 +/- 17 nM to 4505 +/- 403 nM for inhibition of synaptic currents, suggesting that this phenomenon may underlie tolerance. Presynaptic expression of dominant negative G-protein-coupled-receptor kinase (GRK2-Lys220Arg) or beta-arrestin (319-418) reduced the desensitization produced by 18- to 24-h pretreatment with 100 nM, Win55,212-2 suggesting that desensitization followed the prototypical pathway for G-protein-coupled receptors. Prolonged treatment with Win55,212-2 produced a modest increase in the EC(50) for adenosine inhibition of synaptic transmission and pretreatment with cyclopentyladenosine produced a slight increase in the EC(50) for Win55,212-2, suggesting a reciprocal ability to produce heterologous desensitization. The long term changes in synaptic function that accompany chronic cannabinoid exposure will be an important factor in evaluating the therapeutic potential of these drugs and will provide insight into the role of the endocannabinoid system. PMID- 11854428 TI - Distinct effects of different calcium-mobilizing agents on cell death in NG108-15 neuroblastoma X glioma cells. AB - The effects of different calcium-mobilizing agents on cell death were characterized in NG108-15 neuroblastoma x glioma hybrid cells. Carbonyl cyanide p trifluoromethoxyphenylhydrazone (FCCP) increased the cytosolic Ca(2+) concentration ([Ca(2+)](i)) and caused cell death. Thapsigargin (TG) not only increased the [Ca(2+)](i) and caused cell death but also induced neurite outgrowth via activation of phospholipase A(2) and cytochrome P450 epoxygenase. In contrast, bradykinin increased the [Ca(2+)](i), but had no effect on cell morphology or cell death. Cell death occurred by two different mechanisms, one of which was caspase-3-dependent and the other caspase-3-independent. Caspase-3 activation was Ca(2+)-dependent, whereas neurite outgrowth was Ca(2+) independent. TG- or FCCP-induced caspase-3 activation occurred at the same time, but the cell death induced by TG was delayed. TG treatment did not enhance the generation of nitric oxide or cAMP or secretion of glial-derived neurotrophic factor or neurotrophin-3, but activated sphingosine kinase. Furthermore, inhibition of sphingosine kinase accelerated TG-induced cell death, and exogenous sphingosine 1-phosphate (S1P) protected cells from FCCP-induced cell death by about 60%. These results indicate that, in these cells, depletion of intracellular nonmitochondrial or mitochondrial Ca(2+) stores causes cell death, that TG activates phospholipase A(2) and sphingosine kinase, and that arachidonic acid induces neurite outgrowth, whereas S1P delays cell death. PMID- 11854429 TI - Midazolam oxidation by cytochrome P450 3A4 and active-site mutants: an evaluation of multiple binding sites and of the metabolic pathway that leads to enzyme inactivation. AB - Midazolam (MDZ) oxidation by recombinant CYP3A4 purified from Escherichia coli and 30 mutants generated at 15 different substrate recognition site positions has been studied to determine the role of individual residues in regioselectivity and to investigate the possible existence of multiple binding sites. Initial results showed that oxidation of MDZ by CYP3A4 causes time- and concentration-dependent enzyme inactivation with K(I) and k(inact) values of 5.8 microM and 0.15 min(-1), respectively. The different time courses of MDZ hydroxylation by mutants that predominantly formed 1'-OH MDZ as opposed to 4-OH MDZ provided strong evidence that the 1'-OH MDZ pathway leads to CYP3A4 inactivation. Correlational analysis of 1'-OH formation versus 4-OH formation by the mutants supports the inference that the two metabolites result from the binding of MDZ at two separate sites. Thus, substitution of residues Phe-108, Ile-120, Ile-301, Phe-304, and Thr-309 with a larger amino acid caused an increase in the ratio of 1'-OH/4-OH MDZ formation, whereas substitution of residues Ser-119, Ile-120, Leu-210, Phe-304, Ala-305, Tyr-307, and Thr-309 with a smaller amino acid decreased this ratio. Kinetic analyses of nine key mutants revealed that the alteration in regioselectivity is caused by a change in kinetic parameters (V(max) and K(M)) for the formation of both metabolites in most cases. The study revealed the role of various active-site residues in the regioselectivity of MDZ oxidation, identified the metabolic pathway that leads to enzyme inactivation, and provided an indication that the two proposed MDZ binding sites in CYP3A4 may be partially overlapping. PMID- 11854430 TI - Regulation of pulmonary and hepatic cytochrome P4501A expression in the rat by hyperoxia: implications for hyperoxic lung injury. AB - Supplemental oxygen therapy is frequently used in the treatment of pulmonary insufficiency, as is encountered in premature infants, and in patients with acute respiratory distress syndrome. However, hyperoxia causes lung damage in experimental animals and may do so in humans. Cytochrome P4501A enzymes have been implicated in hyperoxic lung injury. In this study, we investigated the mechanisms of CYP1A1 regulation by hyperoxia and tested the hypothesis that aryl hydrocarbon receptor (AHR)-dependent mechanisms contribute to induction of CYP1A1 and that modulation of CYP1A by hyperoxia may have implications for lung injury. Exposure of adult male Sprague-Dawley rats to hyperoxia for 24 to 48 h led to increased expression of pulmonary CYP1A1 enzyme, which was preceded by enhancement of the corresponding mRNA, followed by decline of induction at 60 h, when the animals displayed severe respiratory distress and lung inflammation. Similarly, hepatic CYP1A1/1A2 mRNAs were markedly induced between 24 and 48 h of hyperoxia, with induction declining by 60 h. Electrophoretic mobility shift assays (EMSA) and experiments with AHR (-/-) mice indicated that AHR-dependent mechanisms contributed to CYP1A induction. The AHR (-/-) mice were refractory to CYP1A1 induction by hyperoxia and were more sensitive to lung injury than wild type mice. Lungs of hyperoxic rats showed increase in the expression of CYP1A1 in airway epithelial cells, type II pneumocytes, and endothelial cells. In conclusion, our results suggest that induction of CYP1A1 by hyperoxia is mediated by AHR-dependent mechanisms and that modulation of CYP1A enzymes by hyperoxia may have implications for hyperoxic lung injury. PMID- 11854431 TI - Multiple interference of anthracyclines with mitochondrial creatine kinases: preferential damage of the cardiac isoenzyme and its implications for drug cardiotoxicity. AB - Anthracyclines are among the most efficient drugs of cancer chemotherapy, but their use is limited by a significant risk of cardiotoxicity, which is still far from being understood. This study investigates whether impairment of mitochondrial creatine kinase (MtCK), a key enzyme in cellular energy metabolism, could be involved in anthracycline cardiotoxicity. We have analyzed the effects of three anthracyclines, doxorubicin, daunorubicin, and idarubicin, on two MtCK isoenzymes, sarcomeric/cardiac sMtCK and ubiquitous uMtCK, from human and chicken. Using surface plasmon resonance, gel filtration, and enzyme assays, we have quantified properties that are of basic importance for MtCK functioning in vivo: membrane binding, octameric state, and enzymatic activity. Anthracyclines significantly impaired all three properties with differences in dose-, time-, and drug-dependence. Membrane binding and enzymatic activity were already affected at low anthracycline concentrations (5-100 microM), indicating high clinical relevance. Effects on membrane binding were immediate, probably because of competitive binding of the drug to cardiolipin. In contrast, dissociation of MtCK octamers into dimers, enzymatic inactivation and cross-linking occurred only after hours to days. Different protection assays suggest that the deleterious effects were caused by oxidative damage, mainly affecting the highly susceptible MtCK cysteines, followed by generation of free oxygen radicals at higher drug concentrations. Enzymatic inactivation occurred mainly at the active site and involved Cys278, as indicated by experiments with protective agents and sMtCK mutant C278G. All anthracycline effects were significantly more pronounced for sMtCK than for uMtCK. These in vitro results suggest that sMtCK damage may play a role in anthracycline cardiotoxicity. PMID- 11854432 TI - Endothelin-1 protects ovarian carcinoma cells against paclitaxel-induced apoptosis: requirement for Akt activation. AB - Endothelin-1 (ET-1) is a powerful mitogenic peptide produced by different tumors. In ovarian carcinoma cells, ET-1 acts as an autocrine growth factor, selectively through ET(A) receptor (ET(A)R), which is predominantly expressed in tumor cells. The aim of this study was to examine whether ET-1 plays a role in the sensitivity of three ovarian carcinoma cell lines (OVCA 433, HEY, and SK-OV-3) to apoptosis induced by two different stimuli. Our results demonstrated that the addition of ET-1 markedly inhibited serum withdrawal and paclitaxel-induced apoptosis in a concentration-dependent manner, as demonstrated by Annexin-V assay, sub-G(1) peak in DNA content histograms, internucleosomal DNA fragmentation, and terminal deoxynucleotidyl transferase-mediated dUTP biotin nick-end labeling method. Pretreatment of the cells with an ET(A)R antagonist, BQ 123, reversed the ET-1 induced protective effect. Paclitaxel-induced apoptosis resulted in the phosphorylation of Bcl-2 that was suppressed by the addition of ET-1. Further analysis of the signaling pathway demonstrated that ET-1 stimulated Akt activation. The phosphatidylinositol 3-kinase (PI3-K) inhibitor wortmannin blocked ET-1-induced Akt phosphorylation. Inhibition of ET-1-stimulated mitogen activated protein kinase activity did not affect ET-1 protection from paclitaxel mediated apoptosis. Moreover, BQ 123 blocked the Akt-mediated pathway activated by ET-1, sensitizing ovarian carcinoma cells to paclitaxel treatment. These results establish a novel role for ET-1 in determining protection of ovarian carcinoma cells against paclitaxel-induced apoptosis through Bcl-2-dependent and PI3-K-mediated Akt pathways and suggest that ET-1 and ET(A)R could represent important targets for anticancer therapy. PMID- 11854433 TI - Channel blockers acting at N-methyl-D-aspartate receptors: differential effects of mutations in the vestibule and ion channel pore. AB - A large number of structurally diverse compounds act as open-channel blockers of NMDA receptors. They may share discrete or overlapping binding sites within the channel. In this study, the effects of mutations in and around the membrane spanning and pore-forming regions of NMDA receptor subunits were studied with three blockers, MK-801, memantine, and TB-3-4, using recombinant NMDA receptors expressed in Xenopus laevis oocytes. Mutations at the critical asparagine residues in the M2 loop of NR1 and NR2B and at a tryptophan residue in M2 of NR2B reduced block by MK-801, memantine, and TB-3-4. Mutations at residues in the pre M1, M1, M3, post-M3, and post-M4 regions had differential effects on the three blockers. Many mutations in these regions reduced block by MK-801 and TB-3-4 but had no effect on block by memantine. The differential effects on block by memantine and MK-801 are unlikely to be caused by differences in the size of these blockers. Benzyl rings in MK-801 and TB-3-4 may make hydrophobic interactions with aromatic and hydrophobic amino acid residues in the pore. Some mutations in the pre-M1 and M3 regions generated constitutively open channels, characterized by large holding currents. The effects of the various mutants are discussed in the context of models based on the known structure of the pore of the KcsA potassium channel and on previous studies dealing with solvent accessible residues in NMDA receptor subunits as determined by modification after cysteine mutagenesis. PMID- 11854434 TI - Agonist-induced translocation of the kinin B(1) receptor to caveolae-related rafts. AB - The kallikrein-kinin system, activated during inflammatory conditions and the regulation of specific cardiovascular and renal functions, includes two G protein coupled receptors for bradykinin (BK)-related peptides. The B(1) receptor (B(1)R) subtype is not believed to undergo agonist-induced phosphorylation and endocytosis. A conjugate made of the rabbit B(1)R fused with the yellow variant of green fluorescent protein (YFP) was expressed in mammalian cells. In COS-1 or human embryonic kidney (HEK) 293 cells, the construction exhibited a nanomolar affinity for the agonist radioligand [(3)H]Lys-des-Arg(9)-BK or the antagonist ligand [(3)H]Lys-[Leu(8)]des-Arg(9)-BK and a pharmacological profile virtually identical to that of wild-type B(1)R. Lys-des-Arg(9)-BK stimulation of HEK 293 cells stably expressing B(1)R-YFP but not stimulation of untransfected cells released [(3)H]arachidonate in a phospholipase A(2) assay. B(1)R-YFP was visualized as a continuous labeling of the plasma membranes in stably transfected HEK 293 cells (confocal microscopy). Addition of Lys-des-Arg(9)-BK (1-100 nM) rapidly concentrated the receptor-associated fluorescence into multiple aggregates that remained associated with the plasma membrane (no significant internalization) and colocalized with caveolin-1. This reaction was slowly reversible upon agonist washing at 37 degrees C and prevented pretreatment with a B(1)R antagonist. beta-Cyclodextrin treatment, which extracts cholesterol from membranes and disrupts caveolae-related rafts, prevented agonist-induced redistribution of B(1)R-YFP but not the PLA(2) activation mediated by this receptor. The agonist radioligand copurified with caveolin-1 to a greater extent than the tritiated antagonist in buoyant fractions of HEK 293 cells treated with the ligands. Agonist-induced cellular translocation of the kinin B(1)R to caveolae-related rafts without endocytosis is a novel variation on the theme of G protein-coupled receptor adaptation. PMID- 11854435 TI - Caffeic acid phenethyl ester and curcumin: a novel class of heme oxygenase-1 inducers. AB - Heme oxygenase-1 (HO-1) is a redox-sensitive inducible protein that provides efficient cytoprotection against oxidative stress. Curcumin, a polyphenolic natural compound that possesses anti-tumor and anti-inflammatory properties, has been reported recently to induce potently HO-1 expression in vascular endothelial cells (Free Rad Biol Med 28:1303-1312, 2000). Here, we extend our previous findings by showing that caffeic acid phenethyl ester (CAPE), another plant derived phenolic agent, markedly increases heme oxygenase activity and HO-1 protein in astrocytes. The effect seems to be related to the peculiar chemical structures of curcumin and CAPE, because analogous antioxidants containing only portions of these two molecules were totally ineffective. At a final concentration of 30 microM, both curcumin and CAPE maximally up-regulated heme oxygenase activity while promoting marked cytotoxicity at higher concentrations (50-100 microM). Similar results were obtained with Curcumin-95, a mixture of curcuminoids commonly used as a dietary supplement. Incubation of astrocytes with curcumin or CAPE at concentrations that promoted maximal heme oxygenase activity resulted in an early increase in reduced glutathione followed by a significant elevation in oxidized glutathione contents. A curcumin-mediated increase in heme oxygenase activity was not affected by the glutathione precursor and thiol donor N-acetyl-L-cysteine. These data suggest that regulation of HO-1 expression by polyphenolic compounds is evoked by a distinctive mechanism which is not necessarily linked to changes in glutathione but might depend on redox signals sustained by specific and targeted sulfydryl groups. This study identifies a novel class of natural substances that could be used for therapeutic purposes as potent inducers of HO-1 in the protection of tissues against inflammatory and neurodegenerative conditions. PMID- 11854436 TI - Differential response of estrogen receptors alpha and beta to SP500263, a novel potent selective estrogen receptor modulator. AB - We determined the differential response of a novel SERM, SP500263, on estrogen receptor (ER) alpha and the more recently cloned ER-beta. Because of the high homology of amino acid residues in the ligand-binding domain of ER-alpha and ER beta, we were not surprised to find that SP500263 binds to both ERs equally well. In contrast, SP500263 acts as a strong estrogen agonist in a strictly ER-alpha specific manner in U2OS osteosarcoma cell lines blocking the production of interleukin (IL) 6 and granulocyte macrophage colony-stimulating factor. SP500263 also blocked IL-6 production in primary bone cells. The mechanism of this inhibition is different from the classic estrogen stimulation involving an estrogen response element (ERE). SP500263 does not activate gene expression through an ERE. In contrast to the results observed in U2OS cells, SP500263 acts as a strong estrogen antagonist in an MCF-7 breast cancer proliferation assay. Therefore, SP500263 is a member of a series of next-generation SERMs with functional selectivity toward ER-alpha and a mixed agonist/antagonist profile in a bone cell assay versus a breast cancer assay. The panel of assays described herein allow for the development of receptor-specific ligands that may be further developed into novel pharmaceuticals with an improved profile for the treatments of osteoporosis and breast cancer. PMID- 11854437 TI - Caspase-dependent cleavage of carbamoyl phosphate synthetase II during apoptosis. AB - Carbamoyl phosphate synthetase II (CPSII) is part of carbamoyl phosphate synthetase/aspartate transcarbamoylase/dihydroorotase (CAD), a multienzymatic protein required for the de novo synthesis of pyrimidine nucleotides and cell growth. Herein, we identify CAD as a substrate for caspase-3 degradation in both in vitro and in vivo models of apoptosis. Withdrawal of interleukin-3 or incubation with staurosporine (STS) or doxorubicin (Dox) resulted in proteolytic cleavage of CAD in a myeloid precursor cell line (32D) or in a cell line over expressing CAD. The rapid decline in the CPSII activity paralleled the degradation of CAD and preceded the appearance of Annexin-V-stained apoptotic cells and DNA fragmentation. These events correlated closely with the activation of caspase-3 in these cells and were prevented by the cell-permeable caspase inhibitor N-benzyloxycarbonyl-Asp-Glu-Val-Asp fluoromethyl ketone. Moreover, the incubation of purified CAD with recombinant caspase-3 in vitro generated CAD fragments that were similar to those obtained in vivo. Edman sequencing revealed that two of the major caspase-3 cleavage sites occurred at the sequences EAVD/G and VACD/G within the catalytic (B2) and allosteric (B3) domains of CAD, thus providing a potential mechanism for the rapid inactivation of CPSII during apoptosis. Consistent with this, an enhanced loss of the intracellular pyrimidines (UTP and CTP) was observed in response to STS or DOX-induced apoptosis. Therefore, these studies show that CAD is a novel target for caspase dependent regulation during apoptosis and suggest that the selective inactivation of pyrimidine nucleotide synthesis accompanies the process of apoptosis. PMID- 11854438 TI - Activation of cAMP-dependent protein kinase suppresses the presynaptic cannabinoid inhibition of glutamatergic transmission at corticostriatal synapses. AB - In a previous study, we showed that type 1 cannabinoid (CB(1)) receptor activation substantially depresses the corticostriatal glutamatergic transmission onto striatal neurons in the brain slice preparation. We now report that the adenylyl cyclase activator forskolin and cAMP analog (S)-p-8-(4-chlorophenythil) adenosine-3',5'-monophosphorothioate (Sp-8-CPT-cAMPS) strongly suppressed the synaptic depression induced by cannabimimetic aminoalkylindole, WIN 55,212-2. Application of the cAMP-dependent protein kinase (PKA) inhibitor KT5720 alone had no consistent effect on basal synaptic transmission but the synaptic enhancement elicited by forskolin was blocked. In addition, pretreatment of striatal slices with either KT5720 or another PKA inhibitor, H89, completely abolished the attenuation by forskolin on WIN 55,212-2-induced synaptic depression. The effect of forskolin on CB(1) receptor function was still observed in a low Ca(2+) bathing solution, suggesting that the forskolin's action is not attributable to its ability to saturate the presynaptic transmitter release processes. The possibility that forskolin acted by increasing CB(1) receptor phosphorylation was confirmed by demonstrating that the serine-phosphorylated component with CB(1) receptors was significantly increased after forskolin treatment. This forskolin effect was markedly attenuated in the presence of KT5720. Moreover, the activation of beta-adrenergic receptors by isoproterenol mimics forskolin to elicit a PKA-dependent inhibition of CB(1) receptor function. Together, these observations indicate that the presynaptic inhibitory action of CB(1) receptors at corticostriatal synapses could be negatively regulated by cAMP/PKA-mediated receptor phosphorylation. This effect of PKA may play a functional role in fine tuning glutamatergic transmission at corticostriatal synapses. PMID- 11854439 TI - Functional analysis of six different polymorphic CYP1B1 enzyme variants found in an Ethiopian population. AB - Cytochrome P450 1B1 (CYP1B1) is an extrahepatic enzyme of potential importance for the metabolism of estrogen and for metabolic activation of environmental carcinogens. We investigated an Ethiopian population for functional polymorphisms in the CYP1B1 gene using genomic DNA sequencing and detected three novel single nucleotide polymorphisms (SNPs). One of these (4360C-->G in exon 3) is present at a frequency of 7% and causes an Ala443Gly amino acid substitution. In addition, the four described previously missense mutations Arg48Gly, Ala119Ser, Leu432Val, and Asn453Ser were found with frequencies of 51, 50, 53, and 2%, respectively, yielding a total of 32 possible CYP1B1 haplotypes. Allele-specific PCR methods for haplotype analysis were developed and seven different CYP1B1 alleles were found: CYP1B1*1, *2, *3, *4, *5, *6, and *7 with frequencies of 8, 37, 39, 2, 0.7, 6, and 7%, respectively. The functional properties of different forms of CYP1B1, as well as of the Leu432Val + Asn453Ser and Leu432Val + Ala443Gly variants, were evaluated after heterologous expression of the corresponding cDNAs in Saccaromyces cerevisiae. The results revealed that CYP1B1.6 and CYP1B1.7, having the amino acid substitutions Arg48Gly, Ala119Ser, and Leu432Val in common, exhibited altered kinetics with significantly increased apparent K(m) and lowered V(max) values for both the 2- and 4-hydroxylation of 17 beta-estradiol, whereas the other constructs were indistinguishable from the CYP1B1.1 enzyme. The results emphasize the necessity of a complete haplotype analysis of enzyme variants for evaluation of functional consequences in vivo and for analyses of genetic polymorphisms in relation to, for example, cancer incidence. PMID- 11854440 TI - Intracellular domains of NR2 alter calcium-dependent inactivation of N-methyl-D aspartate receptors. AB - At central excitatory synapses, the transient elevation of intracellular calcium reduces N-methyl-D-aspartate (NMDA) receptor activity. Such 'calcium-dependent inactivation' is mediated by interactions of calcium/calmodulin and alpha-actinin with the C terminus of NMDA receptor 1 (NR1) subunit. However, inactivation is also NR2-subunit specific, because it occurs in NR2A- but not NR2C-containing receptors. We examined the molecular basis for NR2-subunit specificity using chimeric and mutated NMDA receptor subunits expressed in HEK293 cells. We report that the intracellular loop immediately distal to the pore-forming P-loop M2 (M2 3 loop), as well as a short region in the C terminus, are involved in NR2-subunit specificity. Within the M2-3 loop, substitution of residue 619 in NR2A (valine) for the corresponding NR2C residue (isoleucine) reduced inactivation without affecting calcium permeability of the channel. In contrast, a Q620E mutation in NR2A reduced the relative calcium permeability without altering inactivation. Mutation of three serine/threonine residues in the M2-3 loop also reduced inactivation, as did substitution of the intracellular C terminus of NR2A for NR2C. We speculate that the M2-3 loop of NR2 modulates calcium-dependent inactivation by interacting with the NR1 C terminus, a region known to be essential for inactivation. PMID- 11854441 TI - Adenine nucleotide-induced activation of adenosine A(2B) receptors expressed in Xenopus laevis oocytes: involvement of a rapid and localized adenosine formation by ectonucleotidases. AB - We recently demonstrated that extracellular ATP effectively activates adenosine (Ade) A(2B) receptors indirectly through a localized rapid conversion to Ade by ectonucleotidases on the membrane surface of C6Bu-1 rat glioma cells. These responses were observed even in the presence of adenosine deaminase (ADA). Here, we demonstrate that such responses indeed occur in A(2B) receptor-expressing Xenopus laevis oocytes, which possess endogenous ectonucleotidase activity. In oocytes coexpressing the A(2B) receptor and cystic fibrosis transmembrane conductance regulator (CFTR), Ade induced a concentration-dependent increase in a cyclic AMP-activated CFTR current, a response that was inhibited by the P1 antagonist xanthine-amine congener (XAC). A brief application of ATP and beta,gamma-methylene ATP (beta,gamma-MeATP) also induced the CFTR current in a manner similar to that seen with Ade. Among several nucleotide agonists, ADP, AMP, and adenosine-5'-O-(3-thio)triphosphate induced the CFTR current. Although adenine nucleotide-induced CFTR currents were inhibited by XAC, they were highly resistant to ADA treatment; 5 U/ml ADA was required for inhibition of adenine nucleotide-induced CFTR current, whereas 1 U/ml ADA was sufficient to abolish the Ade-induced response. In addition, the ecto-5'-nucleotidase inhibitor alpha,beta methylene ADP markedly inhibited the beta,gamma-MeATP-induced response but not the Ade-induced one. These results support our hypothesis that adenine nucleotides are rapidly and locally converted into Ade on the membrane surface, resulting in the activation of A(2B) receptors. PMID- 11854442 TI - Regulation of cyclooxygenase-2 expression by phospholipase D in human amnion derived WISH cells. AB - Prostaglandins (PGs) are known to play a key role in the initiation of labor, but the mechanisms regulating their synthesis in amnion are largely unknown. In this study, the regulatory mechanisms for PGE(2) production during phospholipase D (PLD) and p38-dependent activation of WISH cells were investigated. We found that the stimulation of WISH cells with interleukin (IL)-1 beta elicited dose dependent synthesis of cyclooxygenase-2 (COX-2) mRNA, protein, and their products, PGE(2). Moreover, the treatment of [(3)H]myristate-labeled cells in the presence of 1-butanol caused the dose-dependent formation of [(3)H]phosphatidylbutanol (PBt), a product specific to PLD activity. Pretreating the cells with 1-butanol and Ro 31-8220 inhibited the IL-1 beta-induced COX-2 expression, but 3-butanol did not affect this response. In addition, evidence that PLD was involved in the stimulation of COX-2 expression was provided by the observations that COX-2 expression was stimulated by the dioctanoyl phosphatidic acid (PA) and that the prevention of PA dephosphorylation by 1-propranolol potentiated COX-2 expression by IL-1 beta. Moreover, IL-1 beta stimulation of the cells caused the phosphorylation of p38 and extracellular signal-regulated kinase (ERK), and IL-1 beta-induced COX-2 expression was inhibited by the pretreatment of WISH cells with a p38 inhibitor, in contrast ERK upstream inhibitor had no effect. Furthermore, Ro 31-8220 inhibited IL-1 beta-induced p38 phosphorylation but not ERK phosphorylation. The results of this study indicate that in human amnion cells, IL-1 beta might activate PLD through an upstream protein kinase C to elicit p38 and finally induce COX-2 expression. PMID- 11854443 TI - Vesnarinone causes oxidative damage by inhibiting catalase function through ceramide action in myeloid cell apoptosis. AB - Vesnarinone is an effective inotropic agent for treating congestive heart failure, but its clinical usage is restricted because of the severe side effect of agranulocytosis. In myeloid HL-60 cells, vesnarinone increased the intracellular content of a proapoptotic lipid mediator, ceramide, in a time- and dose-dependent manner. Vesnarinone-induced apoptosis was significantly enhanced by simultaneous treatment with a cell-permeable N-acetyl sphingosine (C2 ceramide). Treatment with neither vesnarinone, C2-ceramide, nor simultaneously with vesnarinone and C2-ceramide caused a marked increase of reactive oxygen intermediates (ROI) generation measured by the 2',7'-dichlorofluorescin method. However, oxidative damage judged by the production of lipid peroxidates and the nitroblue tetrazolium-reducing ability were enhanced more significantly by simultaneous treatment with vesnarinone and C2-ceramide than by vesnarinone alone. Moreover, vesnarinone inhibited catalase function both at the protein and activity level, and this inhibition was synergistically enhanced by C2-ceramide, and vesnarinone-induced oxidative damage and apoptosis were significantly suppressed by treatment of HL-60 cells with purified catalase. C2-ceramide enhanced vesnarinone-induced inhibition of the ROI-scavenging enzyme catalase at the levels of protein and activity in HL-60 cells; in contrast, however, vesnarinone did not induce ceramide generation, oxidative damage, or catalase depletion in HL-60/ves cells, where vesnarinone could not induce apoptosis. Taken together, the results suggest that vesnarinone induces myeloid cell apoptosis by increasing oxidative damage via ceramide-induced inhibition of catalase function. PMID- 11854444 TI - Activation and inhibition of G proteins by lipoamines. AB - We have previously shown that alkyl-substituted amino acid derivatives directly activate G(i/o) proteins. N-Dodecyl-N(alpha),N(epsilon)-(bis-l-lysinyl)-l-lysine amide (FUB132) is a new representative of this class of compounds with increased efficacy. Here, we characterized the molecular mechanism of action of this class of compounds. FUB132 and its predecessor FUB86 were selective receptomimetics for G(i/o) because they stimulated the guanine nucleotide exchange reaction of purified G(i/o) as documented by an increased rate of GDP release, GTP gamma S binding, and GTP hydrolysis. In contrast to the receptomimetic peptide mastoparan, stimulation of G proteins by lipoamines required the presence of neither G beta gamma-dimers nor lipids. On the contrary, G beta gamma-dimers suppressed the stimulatory effect of FUB132. The stimulation of G(i/o) by lipoamines and by mastoparan was not additive. A peptide derived from the C terminus of G alpha(o3), but not a corresponding G alpha(q)-derived peptide, quenched the FUB132-induced activation of G alpha(o). In membranes prepared from human embryonic kidney 293 cells that stably expressed the G(i/o)-coupled human A(1)-adenosine receptor, lipoamines impeded high-affinity agonist binding. In contrast, antagonist binding was not affected. We conclude that alkyl-substituted amines target a site, most likely at the C terminus of G alpha(i/o)-subunits, that is also contacted by receptors. However, because G beta gamma-dimers blunt rather than enhance their efficacy, their mechanism of action differs fundamentally from that of a receptor. Thus, despite their receptomimetic effect in vitro, alkyl-substituted amines and related polyamines are poor direct G protein activators in vivo. In the presence of G beta gamma, they rather antagonize G protein-coupled receptor signaling. PMID- 11854445 TI - Identification of ligand-binding regions of P-glycoprotein by activated pharmacophore photoaffinity labeling and matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry. AB - Energy dependent efflux pumps confer resistance to anticancer, antimicrobial, and antiparasitic drugs. P-glycoprotein (Pgp, ABCB1) mediates resistance to a broad spectrum of antitumor drugs. Compounds that themselves are nontoxic to cells have been shown to act as inhibitors of Pgp. The mechanism of binding and transport of low-molecular-mass ligands by Pgp is still incompletely understood. This study introduces a series of propafenone-related photoaffinity ligands, which combine high specificity and selectivity for Pgp with high labeling efficiency. Molecules are intrinsically photoactivatable in the arylcarbonyl group, which represents a pharmacophoric substructure for this group of ligand molecules. A detailed study of the structure-activity relationship for this type of photoligand is presented. In subsequent experiments, these ligands were used to characterize the drug binding domain of propafenone-type analogs. Matrix-assisted laser desorption/ionization-time-of-flight (MALDI-TOF) mass spectrometry shows that propafenone-type ligands preferentially label fragments assigned to putative transmembrane segments 3, 5, 6, 8, 10, 11, and 12. Labeled fragments are also identified in a highly charged region of 15 amino acids in the second cytoplasmic loop. This region corresponds to the so-called EAA-like motif, which has been proposed to play a role in the interaction between transmembrane domain and nucleotide binding domain of peroxisomal ATP-binding cassette transporters. In addition, a region in cytoplasmic loop 3 and between TM12 and the N terminus of the Walker A sequence of NBD2 are labeled by the ligands. Therefore, a number of confined protein regions contribute to the drug-binding domain of propafenone type analogs. PMID- 11854446 TI - Synthesis and evaluation of a new class of nifedipine analogs with T-type calcium channel blocking activity. AB - We have synthesized a novel series of 18 dialkyl 1,4-dihydro-4-(2'alkoxy-6' pentadecylphenyl)-2,6-dimethyl-3,5 pyridine dicarboxylates from anacardic acid, a natural compound found in cashew nut shells, and investigated their blocking action on L- and T-type calcium channels transiently expressed in tSA-201 cells. The IC(50) values for L-type calcium channel block obtained with the series ranged from 1 to approximately 40 microM, with higher affinities being favored by increasing the size of the alkoxy group on the 4-phenyl ring and ester substituent in the 3,5 positions. A detailed analysis of the strongest L-type channel blocker of the series (PPK-12) revealed that block was poorly reversible and mediated an apparent speeding of the time course of inactivation. Moreover, in the presence of PPK-12, the midpoint of the steady state inactivation curve was shifted by 20 mV toward more hyperpolarized potentials, resulting in an increase in blocking efficacy at more depolarized holding potentials. Surprisingly, PPK-12 blocked T- and L-type calcium channels with similar affinities. One of the weakest L-type channel inhibitors (PPK-5) exhibited a T type channel affinity that was similar to that seen with PPK-12, resulting in a 40-fold selectivity of PPK-5 for T- over L-type channels. Thus, dialkyl 1,4 dihydro-4-(2'alkoxy-6'-pentadecylphenyl)-2,6-dimethyl-3,5 pyridine dicarboxylates may serve as excellent candidates for the development of T-type calcium-channel specific blockers. PMID- 11854447 TI - Inactivation of the Saccharomyces cerevisiae SKY1 gene induces a specific modification of the yeast anticancer drug sensitivity profile accompanied by a mutator phenotype. AB - The therapeutic potential of the highly active anticancer agent cisplatin is severely limited by the occurrence of cellular resistance. A better understanding of the molecular pathways involved in cisplatin-induced cell death could potentially indicate ways to overcome cellular unresponsiveness to the drug and thus lead to better treatment results. We used the budding yeast Saccharomyces cerevisiae as a model organism to identify and characterize novel genes involved in cisplatin-induced cell kill, and found that SKY1 (SR-protein-specific kinase from budding yeast) is a cisplatin sensitivity gene whose disruption conferred cisplatin resistance. In cross-resistance studies, we observed resistance of yeast sky1 Delta cells (i.e., cells from which the SKY1 gene had been disrupted) to cisplatin, carboplatin (but not oxaliplatin), doxorubicin and daunorubicin, and hypersensitivity to cadmium chloride and 5-fluorouracil. Furthermore, these cells did not display reduced platinum accumulation, DNA platination or doxorubicin accumulation, indicating that the resistance is unrelated to decreased drug import or increased drug export. Based on the modification of the anticancer drug sensitivity profile and our finding that sky1 Delta cells display a mutator phenotype, we propose that Sky1p might play a significant role in specific repair and/or tolerance pathways. Disruption of the S. cerevisiae SKY1 gene would thus result in deregulation of such mechanisms and, consequently, lead to altered drug sensitivity. PMID- 11854449 TI - Non-ahr gene susceptibility Loci for porphyria and liver injury induced by the interaction of 'dioxin' with iron overload in mice. AB - Among the actions of 2,3,7,8-tetrachlorodibenzo-p-dioxin (dioxin) in mice is the induction of hepatic porphyria. This is similar to the most common disease of this type in humans, sporadic porphyria cutanea tarda (PCT). Evidence is consistent with the actions of dioxin being mediated through binding to the aryl hydrocarbon receptor (AHR) with different Ahr alleles in mouse strains apparently accounting for differential downstream gene expression and susceptibility. However, studies of dioxin-induced porphyria and liver injury indicate that the mechanisms must involve interactions with other genes, perhaps associated with iron metabolism. We performed a quantitative trait locus (QTL) analysis of an F(2) cross between susceptible C57BL/6J (Ahr(b1) allele) and the highly resistant DBA/2 (Ahr(d) allele) strains after treatment with dioxin and iron. For porphyria we found QTLs on chromosomes 11 and 14 in addition to the Ahr gene (chromosome 12). Studies with C57BL/6.D2 Ahr(d) mice confirmed that the Ahr(d) allele alone did not completely negate the response. SWR mice are syngenic for the Ahr(d) allele with the DBA/2 strain but are susceptible to porphyria after elevation of hepatic iron. Analysis of SWRxD2 F(2) mice treated with iron and dioxin showed a QTL on chromosome 11, as well as finding other loci on chromosomes 1 (and possibly 9), for both porphyria and liver injury. These findings show for the first time the location of genes, other than Ahr, that modulate the mechanism of hepatic porphyria and injury caused by dioxin in mice. Orthologous loci may contribute to the pathogenesis of human sporadic PCT. PMID- 11854448 TI - Reconstitution of the cytoplasmic interaction between phospholamban and Ca(2+) ATPase of cardiac sarcoplasmic reticulum. AB - Phospholamban (PLN) reversibly inhibits the Ca(2+)-ATPase of cardiac sarcoplasmic reticulum (SERCA2a) through a direct protein-protein interaction, playing a pivotal role in the regulation of intracellular Ca(2+) in heart muscle cells. The interaction between PLN and SERCA2a occurs at multiple sites within the cytoplasmic and membrane domains. Here, we have reconstituted the cytoplasmic protein-protein interaction using bacterially expressed fusion proteins of the cytoplasmic domain of PLN and the long cytoplasmic loop of SERCA2a. We have developed two methods to evaluate the binding of the fusion proteins, one with glutathione-Sepharose beads and the other with a 96-well plate. Essentially the same results were obtained by the two methods. The affinity of the binding (K(D)) was 0.70 microM. The association was inhibited by cAMP-dependent phosphorylation of the PLN fusion protein and by usage of anti-PLN monoclonal antibody. It was also diminished by substitution at the phosphorylation site of PLN of Ser(16) to Asp. These results suggest that PLN can bind SERCA2a in the absence of the membrane domains and that the modifications of the cytoplasmic domain of PLN that activate SERCA2a parallel the disruption of the association between the two fusion proteins. It has been shown that the removal of PLN inhibition of SERCA2a rescues cardiac function and morphology in the mouse dilated cardiomyopathy model. Our assay system can be applied to the screening of novel inotropic agents that remove the inhibition of SERCA2a by PLN, improving the relaxation as well as the contractility of the failing heart. PMID- 11854450 TI - Single-channel pharmacology of mibefradil in human native T-type and recombinant Ca(v)3.2 calcium channels. AB - To study the molecular pharmacology of low-voltage-activated calcium channels in biophysical detail, human medullary thyroid carcinoma (hMTC) cells were investigated using the single-channel technique. These cells had been reported to express T-type whole-cell currents and a Ca(v)3.2 (or alpha 1H) channel subunit. We observed two types of single-channel activity that were easily distinguished based on single-channel conductance, voltage dependence of activation, time course of inactivation, rapid gating kinetics, and the response to the calcium agonist (S)-Bay K 8644. Type II channels had biophysical properties (activation, inactivation, conductance) typical for high-voltage-activated calcium channels. They were markedly stimulated by 1 microM (S)-Bay K 8644, allowing to identify them as L-type channels. The channel termed type I is a low-voltage-activated, small-conductance (7.2 pS) channel that inactivates rapidly and is not modulated by (S)-Bay K 8644. Type I channels are therefore classified as T-type channels. They were strongly inhibited by 10 microM mibefradil. Mibefradil block was caused by changes in two gating parameters: a pronounced reduction in fraction of active sweeps and a slight shortening of the open-state duration. Single recombinant low voltage-activated T-type calcium channels were studied in comparison, using human embryonic kidney 293 cells overexpressing the pore-forming Ca(v)3.2 subunit. Along all criteria examined (mechanisms of block, extent of block), recombinant Ca(v)3.2 interact with mibefradil in the same way as their native counterparts expressed in hMTC cells. In conclusion, the pharmacologic phenotype of these native human T-type channels--as probed by mibefradil--is similar to recombinant human Ca(v)3.2. PMID- 11854451 TI - Increased sensitivity to nicotine-induced seizures in mice expressing the L250T alpha 7 nicotinic acetylcholine receptor mutation. AB - High doses of nicotine, the addictive component of tobacco, induce clonic-tonic seizures in animals. Pharmacological and biochemical data have suggested that alpha 7-containing neuronal nicotinic receptors (nAChRs) contribute to these seizures. To study potential alpha 7 contributions, we examined alpha 7 subunits with a Leu250-to-Thr substitution in the channel domain, which creates a gain-of function mutation. Previous studies have shown that mice homozygous for the alpha 7 L250T mutation (T/T) die shortly after birth, but animals heterozygous for the mutation (+/T) are viable and grow to adulthood. Hippocampal neurons from the +/T mice exhibited altered alpha 7-type currents with increased amplitudes and slower desensitization kinetics, confirming a partial gain of function for the alpha 7 nAChR. We found that +/T mice were more sensitive to the convulsant effects of nicotine compared with their wild-type (+/+) littermates. Furthermore, although their behavior was normal in basal conditions, +/T mice showed a unique nicotine induced phenotype, consisting of head-bobbing and paw-tapping movements. Increased sensitivity to nicotine-induced seizures occurred despite a 60% decline in brain alpha 7 nAChR protein levels. There were no changes in the levels of alpha 4, alpha 5, alpha 6, alpha 7, beta 2, and beta 4 mRNA, or in [(125)I]epibatidine and [(3)H]nicotine binding between +/T and +/+ mice. Recent data from our laboratory show that alpha 7-null mice maintain normal sensitivity to nicotine-induced seizures. Hence, these present findings suggest that alterations in the properties rather than absence of alpha 7 nAChRs might affect the mechanisms underlying the convulsive properties of nicotine. PMID- 11854452 TI - A comparison of experimental, theoretical, and numerical simulation Rayleigh Taylor mixing rates. AB - We present a Rayleigh-Taylor mixing rate simulation with an acceleration rate falling within the range of experiments. The simulation uses front tracking to prevent interfacial mass diffusion. We present evidence to support the assertion that the lower acceleration rate found in untracked simulations is caused, at least to a large extent, by a reduced buoyancy force due to numerical interfacial mass diffusion. Quantitative evidence includes results from a time-dependent Atwood number analysis of the diffusive simulation, which yields a renormalized mixing rate coefficient for the diffusive simulation in agreement with experiment. PMID- 11854453 TI - Endocardial cushion and myocardial defects after cardiac myocyte-specific conditional deletion of the bone morphogenetic protein receptor ALK3. AB - Receptors for bone morphogenetic proteins (BMPs), members of the transforming growth factor-beta (TGFbeta) superfamily, are persistently expressed during cardiac development, yet mice lacking type II or type IA BMP receptors die at gastrulation and cannot be used to assess potential later roles in creation of the heart. Here, we used a Cre/lox system for cardiac myocyte-specific deletion of the type IA BMP receptor, ALK3. ALK3 was specifically required at mid gestation for normal development of the trabeculae, compact myocardium, interventricular septum, and endocardial cushion. Cardiac muscle lacking ALK3 was specifically deficient in expressing TGFbeta2, an established paracrine mediator of cushion morphogenesis. Hence, ALK3 is essential, beyond just the egg cylinder stage, for myocyte-dependent functions and signals in cardiac organogenesis. PMID- 11854454 TI - Light modulation of Rubisco in Arabidopsis requires a capacity for redox regulation of the larger Rubisco activase isoform. AB - The light activation of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) in vivo requires the presence of Rubisco activase, a nuclear-encoded chloroplast protein that consists of two isoforms arising from alternative splicing in most plants. We examined the function of each isoform by characterizing Rubisco activation in transgenic Arabidopsis plants that express only one or both isoforms, as compared with the wild type. In plants expressing only the shorter isoform, Rubisco activity was as high as in the wild type under saturating light, but the activity was not down-regulated at intensities limiting for photosynthesis. In contrast, in plants expressing only the longer isoform, Rubisco activity was down-regulated at limiting light, but the activity was slightly lower and increased much more slowly at saturating light intensities as compared with the wild type. Light regulation of Rubisco similar to that in the wild-type plants was observed in the progeny of a genetic cross of these two transformants in which both isoforms were again present. When the capacity to redox regulate the activity of the larger activase isoform was eliminated by replacement of the critical cysteine residues in the carboxyl-terminal extension unique to this isoform, Rubisco activity in saturating light was similar to the wild type, but the ability of the larger isoform to down-regulate Rubisco activity at limiting light intensities in transgenic plants was almost abolished. These results indicate that the light modulation of Rubisco under limiting light is mainly due to the ability to regulate the activity of Rubisco activase by redox changes in the stroma. PMID- 11854455 TI - Cooperativity of Nkx3.1 and Pten loss of function in a mouse model of prostate carcinogenesis. AB - Mouse models have provided significant insights into the molecular mechanisms of tumor suppressor gene function. Here we use mouse models of prostate carcinogenesis to demonstrate that the Nkx3.1 homeobox gene undergoes epigenetic inactivation through loss of protein expression. Loss of function of Nkx3.1 in mice cooperates with loss of function of the Pten tumor suppressor gene in cancer progression. This cooperativity results in the synergistic activation of Akt (protein kinase B), a key modulator of cell growth and survival. Our findings underscore the significance of interactions between tissue-specific regulators such as Nkx3.1 and broad-spectrum tumor suppressors such as Pten in contributing to the distinct phenotypes of different cancers. PMID- 11854456 TI - A spliceosomal intron in Giardia lamblia. AB - Short introns occur in numerous protist lineages, but there are no reports of intervening sequences in the protists Giardia lamblia and Trichomonas vaginalis, which may represent the deepest known branches in the eukaryotic line of descent. We have discovered a 35-bp spliceosomal intron in a gene encoding a putative [2Fe 2S] ferredoxin of G. lamblia. The Giardia intron contains a canonical splice site at its 3' end (AG), a noncanonical splice site at its 5' end (CT), and a branch point sequence that fits the yeast consensus sequence of TACTAAC except for the first nucleotide (AACTAAC). We have also identified several G. lamblia genes with spliceosomal peptides, including homologues of eukaryote-specific spliceosomal peptides (Prp8 and Prp11), several DExH-box RNA-helicases that have homologues in eubacteria, but serve essential functions in the splicing of introns in eukaryotes, and 11 predicted archaebacteria-like Sm and like-Sm core peptides, which coat small nuclear RNAs. Phylogenetic analyses show the Giardia Sm core peptides are the products of multiple, ancestral gene duplications followed by divergence, but they retain strong similarity to Sm and like-Sm peptides of other eukaryotes. Although we have documented only a single intron in Giardia, it likely has other introns and fully functional, spliceosomal machinery. If introns were added during eukaryotic evolution (the introns-late hypothesis), then these results push back the date of this event before the branching of G. lamblia. PMID- 11854457 TI - The human programmed cell death-2 (PDCD2) gene is a target of BCL6 repression: implications for a role of BCL6 in the down-regulation of apoptosis. AB - BCL6, a gene on chromosome 3 band q27, encodes a Kruppel-type zinc finger transcriptional repressor. Rearrangements of this gene are frequent in various kinds of lymphomas, particularly of the large-cell B-cell type. The BCL6 nuclear phosphoprotein is expressed in a variety of tissues and is up-regulated particularly in lymph node germinal centers. The zinc fingers of BCL6 bind DNA in a sequence-specific manner. To identify targets of the BCL6 repressive effects, we used a VP16-BCL6 fusion protein containing the zinc fingers but devoid of the repressor domains to compete with the binding of endogenous BCL6 in a transiently transfected B-cell line and then performed subtractive hybridization by using a method to selectively amplify sequences that are differentially expressed. We found that the programmed cell death-2 (PDCD2) gene is a target of BCL6 repression. This gene is the human homolog of Rp8, a rat gene associated with programmed cell death in thymocytes. Immunohistochemistry reveals the anticipated inverse relationship between BCL6 and PDCD2 expression in human tonsil. PDCD2 is detectable in cells of the germinal center in areas where there is less BCL6 expression as well as in the mantle zone, where there is little or no BCL6 expression. These results raise the possibility that BCL6 may regulate apoptosis by means of its repressive effects on PDCD2. BCL6 deregulation may lead to persistent down-regulation of PDCD2, reduced apoptosis, and, as a consequence, accumulation of BCL6-containing lymphoma cells. PMID- 11854458 TI - Up-regulation of nitric oxide synthase and modulation of the guanylate cyclase activity by corticotropin-releasing hormone but not urocortin II or urocortin III in cultured human pregnant myometrial cells. AB - The biological actions of corticotropin-releasing hormone (CRH) in the human myometrium during pregnancy and labor are unknown. We hypothesized that CRH may modulate the nitric oxide system, and influence myometrial relaxation/contractility. Incubation of myometrial cells with CRH, but not urocortin II or urocortin III, for 8-16 h significantly induced mRNA and protein expression of endothelial and brain but not inducible nitric oxide synthase (NOS) isoforms. This action resulted in increased activity of soluble guanylate cyclase (GC(s)), demonstrated by the enhanced cGMP-producing capacity of the NO donor, sodium nitroprusside. CRH also caused acute activation of the membrane-bound GC, shown by increased basal or atrial natriuretic peptide (ANP)-stimulated cGMP production. These effects appeared to be mediated via the R1 receptors because the CRH receptor antagonists, astressin and antalarmin but not anti-sauvagine 30, could block them. The acute effects of CRH were significantly reduced by inhibition of protein kinase A (PKA) activity, suggesting it is partially PKA dependent. Activation of protein kinase C (PKC) resulted in significant inhibition of both ANP-and CRH-stimulated cGMP production, suggesting a direct effect of PKC on membrane-bound GC. In conclusion, CRH appears to have a dual effect on myometrial NOS/GC pathway, a short term effect predominantly mediated by PKA, and a long-term effect increasing constitutive NOS expression, mediated by a PKA-independent mechanism. This mechanism could potentially be active during human pregnancy, and, because cGMP stimulates myometrial relaxation, these findings further suggest that during pregnancy CRH primarily activates intracellular signals that contribute to the maintenance of myometrial quiescence. PMID- 11854459 TI - The mur2 mutant of Arabidopsis thaliana lacks fucosylated xyloglucan because of a lesion in fucosyltransferase AtFUT1. AB - Cell walls of the Arabidopsis mutant mur2 contain less than 2% of the wild-type amount of fucosylated xyloglucan because of a point mutation in the fucosyltransferase AtFUT1. The mur2 mutation eliminates xyloglucan fucosylation in all major plant organs, indicating that Arabidopsis thaliana fucosyltransferase 1 (AtFUT1) accounts for all of the xyloglucan fucosyltransferase activity in Arabidopsis. Despite this alteration in structure, mur2 plants show a normal growth habit and wall strength. In contrast, Arabidopsis mur1 mutants that are defective in the de novo synthesis of l-fucose exhibit a dwarfed growth habit and decreased wall strength [Reiter, W. D., Chapple, C. & Somerville, C. R. (1993) Science 261, 1032-1035]. Because the mur1 mutation affects several cell wall polysaccharides, whereas the mur2 mutation is specific to xyloglucan, the phenotypes of mur1 plants appear to be caused by structural changes in fucosylated pectic components such as rhamnogalacturonan II. The normal growth habit and wall strength of mur2 plants casts doubt on hypotheses regarding roles of xyloglucan fucosylation in facilitating xyloglucan cellulose interactions or in modulating growth regulator activity. PMID- 11854460 TI - The interaction between the coactivator dCBP and Modulo, a chromatin-associated factor, affects segmentation and melanotic tumor formation in Drosophila. AB - The development of Drosophila requires the function of the CREB-binding protein, dCBP. In flies, dCBP serves as a coactivator for the transcription factors Cubitus interruptus, Dorsal, and Mad, and as a cosuppressor of Drosophila T cell factor. Current models propose that CBP, through its intrinsic and associated histone acetyltransferase activities, affects transient chromatin changes that allow the preinitiation complex to access the promoter. In this report, we provide evidence that dCBP may regulate the formation of chromatin states through interactions with the modulo (mod) gene product, a protein that is thought to be involved in chromatin packaging. We demonstrate that dCBP and Modulo bind in vitro and in vivo, that mutations in mod enhance the embryonic phenotype of a dCBP mutation, and that dCBP mutations enhance the melanotic tumor phenotype characteristic of mod homozygous mutants. These results imply that, in addition to its histone acetyltransferase activity, dCBP may affect higher-order chromatin structure. PMID- 11854461 TI - Arctic "ozone hole" in a cold volcanic stratosphere. AB - Optical depth records indicate that volcanic aerosols from major eruptions often produce clouds that have greater surface area than typical Arctic polar stratospheric clouds (PSCs). A trajectory cloud-chemistry model is used to study how volcanic aerosols could affect springtime Arctic ozone loss processes, such as chlorine activation and denitrification, in a cold winter within the current range of natural variability. Several studies indicate that severe denitrification can increase Arctic ozone loss by up to 30%. We show large PSC particles that cause denitrification in a nonvolcanic stratosphere cannot efficiently form in a volcanic environment. However, volcanic aerosols, when present at low altitudes, where Arctic PSCs cannot form, can extend the vertical range of chemical ozone loss in the lower stratosphere. Chemical processing on volcanic aerosols over a 10-km altitude range could increase the current levels of springtime column ozone loss by up to 70% independent of denitrification. Climate models predict that the lower stratosphere is cooling as a result of greenhouse gas built-up in the troposphere. The magnitude of column ozone loss calculated here for the 1999--2000 Arctic winter, in an assumed volcanic state, is similar to that projected for a colder future nonvolcanic stratosphere in the 2010 decade. PMID- 11854462 TI - Rapid assembly dynamics of the Escherichia coli FtsZ-ring demonstrated by fluorescence recovery after photobleaching. AB - FtsZ, the major cytoskeletal component of the bacterial cell-division machine, assembles into a ring (the Z-ring) that contracts at septation. FtsZ is a bacterial homolog of tubulin, with similar tertiary structure, GTP hydrolysis, and in vitro assembly. We used green fluorescent protein-labeled FtsZ and fluorescence recovery after photobleaching to show that the E. coli Z-ring is extremely dynamic, continually remodeling itself with a half-time of 30 s. ZipA, a membrane protein involved in cell division that colocalizes with FtsZ, was equally dynamic. The Z-ring of the mutant ftsZ84, which has 1/10 the guanosine triphosphatase activity of wild-type FtsZ in vitro, showed a 9-fold slower turnover in vivo. This finding implies that assembly dynamics are determined primarily by GTP hydrolysis. Despite the greatly reduced assembly dynamics, the ftsZ84 cells divide with a normal cell-cycle time. PMID- 11854463 TI - tRNA-like recognition of group I introns by a tyrosyl-tRNA synthetase. AB - The Neurospora crassa mitochondrial tyrosyl-tRNA synthetase (CYT-18 protein) functions in splicing group I introns by promoting the formation of the catalytically active RNA structure. Previous work suggested that CYT-18 recognizes a conserved tRNA-like structure of the group I intron catalytic core. Here, directed hydroxyl-radical cleavage assays show that the nucleotide-binding fold and C-terminal domains of CYT-18 interact with the expected group I intron cognates of the aminoacyl-acceptor stem and D-anticodon arms, respectively. Further, three-dimensional graphic modeling, supported by biochemical data, shows that conserved regions of group I introns can be superimposed over interacting regions of the tRNA in a Thermus thermophilus TyrRS/tRNA(Tyr) cocrystal structure. Our results support the hypothesis that CYT-18 and other aminoacyl tRNA synthetases interact with group I introns by recognizing conserved tRNA-like structural features of the intron RNAs. PMID- 11854464 TI - Selective imaging of presynaptic activity in the mouse olfactory bulb shows concentration and structure dependence of odor responses in identified glomeruli. AB - More chemicals can be smelled than there are olfactory receptors for them, necessitating a combinatorial representation by somewhat broadly tuned receptors. To understand the perception of odor quality and concentration, it is essential to establish the nature of the receptor repertoires that are activated by particular odorants at particular concentrations. We have taken advantage of the one-to-one correspondence of glomeruli and olfactory receptor molecules in the mouse olfactory bulb to analyze the tuning properties of a major receptor population by high resolution calcium imaging of odor responses selectively in the presynaptic compartment of glomeruli. We show that eighty different olfactory receptors projecting to the dorsal olfactory bulb respond to high concentrations of aldehydes with limited specificity. Varying ensembles of about 10 to 20 receptors encode any particular aldehyde at low stimulus concentrations with high specificity. Even normalized odor response patterns are markedly concentration dependent, caused by pronounced differences in affinity within the aldehyde receptor repertoire. PMID- 11854466 TI - Molecular basis of R-type calcium channels in central amygdala neurons of the mouse. AB - R-type Ca2+ channels play a critical role in coupling excitability to dendritic Ca2+ influx and neuronal secretion. Unlike other types of voltage-sensitive Ca2+ channels (L, N, P/Q, and T type), the molecular basis for the R-type Ca2+ channel is still unclear, thereby limiting further detailed analyses of R-type Ca2+ channel physiology. The prevailing hypothesis is that alpha(1E) (Ca(V)2.3) gene encodes for R-type Ca2+ channels, but the dearth of critical evidence has rendered this hypothesis controversial. Here we generated alpha1E-deficient mice (alpha1E-/-) and examined the status of voltage-sensitive Ca2+ currents in central amygdala (CeA) neurons that exhibit abundant alpha1E expression and R type Ca2+ currents. The majority of R-type currents in CeA neurons were eliminated in alpha1E-/- mice whereas other Ca2+ channel types were unaffected. These data clearly indicate that the expression of alpha1E gene underlies R-type Ca2+ channels in CeA neurons. Furthermore, the alpha1E-/- sign mice exhibited signs of enhanced fear as evidenced by their vigorous escaping behavior and aversion to open-field conditions. These latter findings imply a possible role of alpha1E-based R-type Ca2+ currents in amygdala physiology associated with fear. PMID- 11854465 TI - Quorum-sensing regulators control virulence gene expression in Vibrio cholerae. AB - The production of virulence factors including cholera toxin and the toxin coregulated pilus in the human pathogen Vibrio cholerae is strongly influenced by environmental conditions. The well-characterized ToxR signal transduction cascade is responsible for sensing and integrating the environmental information and controlling the virulence regulon. We show here that, in addition to the known components of the ToxR signaling circuit, quorum-sensing regulators are involved in regulation of V. cholerae virulence. We focused on the regulators LuxO and HapR because homologues of these two proteins control quorum sensing in the closely related luminous marine bacterium Vibrio harveyi. Using an infant mouse model, we found that a luxO mutant is severely defective in colonization of the small intestine. Gene arrays were used to profile transcription in the V. cholerae wild type and the luxO mutant. These studies revealed that the ToxR regulon is repressed in the luxO mutant, and that this effect is mediated by another negative regulator, HapR. We show that LuxO represses hapR expression early in log-phase growth, and constitutive expression of hapR blocks ToxR regulon expression. Additionally, LuxO and HapR regulate a variety of other cellular processes including motility, protease production, and biofilm formation. Together these data suggest a role for quorum sensing in modulating expression of blocks of virulence genes in a reciprocal fashion in vivo. PMID- 11854467 TI - Cell senescence and telomere shortening induced by a new series of specific G quadruplex DNA ligands. AB - Telomeres of human chromosomes contain a G-rich 3'-overhang that adopts an intramolecular G-quadruplex structure in vitro which blocks the catalytic reaction of telomerase. Agents that stabilize G-quadruplexes have the potential to interfere with telomere replication by blocking the elongation step catalyzed by telomerase and can therefore act as antitumor agents. We have identified by Fluorescence Resonance Energy Transfer a new series of quinoline-based G quadruplex ligands that also exhibit potent and specific anti-telomerase activity with IC50 in the nanomolar concentration range. Long term treatment of tumor cells at subapoptotic dosage induces a delayed growth arrest that depends on the initial telomere length. This growth arrest is associated with telomere erosion and the appearance of the senescent cell phenotype (large size and expression of beta-galactosidase activity). Our data show that a G-quadruplex interacting agent is able to impair telomerase function in a tumor cell thus providing a basis for the development of new anticancer agents. PMID- 11854468 TI - A basolateral sorting motif in the MICA cytoplasmic tail. AB - The MHC class I chain-related MICA molecule is a stress-induced, highly polymorphic, epithelia-specific, membrane-bound glycoprotein interacting with the activating NK cell receptor NKG2D and/or gut-enriched Vdelta1-bearing gammadelta T cells. We have previously reported the presence of a MICA transmembrane-encoded short-tandem repeat harboring a peculiar allele, A5.1, characterized by a frame shift mutation leading to a premature intradomain stop codon, thus denying the molecule of its 42-aa cytoplasmic tail. Given that this is the most common population-wide MICA allele found, we set out to analyze the functional consequences of cytoplasmic tail deletion. Here, we show native expression of MICA at the basolateral surface of human intestinal epithelium, the site of putative interaction with intraepithelial T and NK lymphocytes. We then demonstrate, in polarized epithelial cells, that although the full-length MICA protein is sorted to the basolateral membrane, the cytoplasmic tail-deleted construct as well as the naturally occurring A5.1 allele are aberrantly transported to the apical surface. Site-directed mutagenesis identified the cytoplasmic tail-encoded leucine-valine dihydrophobic tandem as the basolateral sorting signal. Hence, the physiological location of MICA within epithelial cells is governed by its cytoplasmic tail, implying impairment in A5.1 homozygous individuals, perhaps relevant to the immunological surveillance exerted by NK and T lymphocytes on epithelial malignancies. PMID- 11854469 TI - Sexually dimorphic expression of estrogen receptor beta in the anteroventral periventricular nucleus of the rat preoptic area: implication in luteinizing hormone surge. AB - Striking sex difference was detected in the expression of estrogen receptor (ER) beta mRNA and protein by nonisotopic in situ hybridization and immunohistochemistry in the anteroventral periventricular nucleus (AVPV) of the rat preoptic area. In females more than in males, a significantly larger number of ERbeta mRNA-positive cells were visualized in the medial-most portion of the AVPV within 50 microm from the ependymal lining of the third ventricle. Rats of 7, 14, 21, 35, and 60 days of age (d 1 = day of birth) showed the sex difference. Orchidectomy of male neonates or estrogen treatment of female pups reversed the brain phenotype when examined on d 14. In the AVPV of adult females, ERalpha immunoreactivity colocalized in 83% of ERbeta mRNA-positive cells. Tyrosine hydroxylase immunoreactivity colocalized in 18% of ERbeta immunoreactive cells in d 21 females. Infusion of an ERbeta antisense oligonucleotide into the third ventricle in the vicinity of the AVPV resulted in significantly longer days of successive estrus and a 50% reduction in the number of ERbeta-immunoreactive cells in the AVPV. These findings provide support for the hypothesis that activation of ERbeta in the AVPV is an important regulatory event in the female typical induction of luteinizing hormone surge by estrogen. PMID- 11854470 TI - Identification in traditional herbal medications and confirmation by synthesis of factors that inhibit cholera toxin-induced fluid accumulation. AB - Kampo formulations are traditional herbal medications used in China and Japan for many centuries to treat diarrheal diseases such as cholera. Our studies were undertaken to identify and verify by chemical synthesis the active components that inhibited cholera toxin (CT), the virulence factor secreted by Vibrio cholerae, the causative agent of cholera. The Kampo formulation, Daio-kanzo-to, inhibited CT activities (i.e., ADP-ribosylation, Chinese hamster ovary cell elongation); in Daio-kanzo-to, Daio (Rhei rhizoma) was responsible for this effect. Among several components purified from Daio extract, rhubarb galloyl tannin, a compound characterized by a polygallate structure, was the most effective. To define the active component, gallate analogues similar to rhubarb galloyl-tannin were synthesized. These gallate compounds inhibited all CT activities including ADP-ribosylation, elongation of Chinese hamster ovary cells, and importantly, fluid accumulation in ileal loops. Thus, Kampo formulations or their gallate components might be effective adjunctive therapy with oral rehydration solution for the severe diarrhea of cholera. PMID- 11854471 TI - Noninvasive quantitative imaging of protein-protein interactions in living subjects. AB - We are developing methods to image molecular and cellular events in living subjects. In this study, we validate imaging of protein-protein interactions in living mice by using bioluminescent optical imaging. We use the well studied yeast two-hybrid system adapted for mammalian cells and modify it to be inducible. We employ the NF-kappaB promoter to drive expression of two fusion proteins (VP16-MyoD and GAL4-ID). We modulate the NF-kappaB promoter through tumor necrosis factor alpha. Firefly luciferase reporter gene expression is driven by the interaction of MyoD and ID through a transcriptional activation strategy. We demonstrate the ability to detect this induced protein-protein interaction in cell culture and image this induced interaction in living mice by using transiently transfected cells. The current approach will be a valuable and potentially generalizable tool to noninvasively and quantitatively image protein protein interactions in living subjects. The approaches validated should have important implications for the study of protein-protein interactions in cells maintained in their natural in vivo environment as well as for the in vivo evaluation of new pharmaceuticals targeted to modulate protein-protein interactions. PMID- 11854473 TI - Homeostatic T cell proliferation in a T cell-dendritic cell coculture system. AB - Naive T cells do not proliferate in normal individuals in the absence of antigen stimulation, but they proliferate spontaneously when T cells are severely depleted. We show here that coculture of syngeneic dendritic cells (DC) with naive T cells expressing a single T cell receptor also results in T cell proliferation in the absence of foreign antigen. As in lymphopenic mice, where T cell proliferation depends upon DC, this response in the coculture system requires interaction of the T cells' T cell receptor with self-peptide-MHCs on DC. This in vitro proliferation also requires soluble factors, including IL-15 secreted by DC, and can be inhibited potently by cell-cell contact with CD4+CD25+ regulatory T cells. The coculture system described may illuminate mechanisms that maintain stable numbers of T cells in normal individuals. PMID- 11854472 TI - Poly(ADP-ribose) polymerase-1 dependence of stress-induced transcription factors and associated gene expression in glia. AB - Poly(ADP-ribose) polymerase-1 (PARP-1, EC ), a nuclear enzyme activated by DNA strand breaks, physiologically participates in DNA repair. Excessive activation of PARP-1 by cellular insults depletes its substrate beta-nicotinamide adenine dinucleotide and ATP, leading to cell death. PARP-1-deficient (PARP-1-/-) mice are protected from several forms of inflammation. In the present study, we demonstrate in PARP-1-/- glial cells a loss of several stress-activated transcription factors as well as decreased expression of genes for cytokines and cellular adhesion molecules. We also show that augmented expression of some of these genes is independent of PARP-1 catalytic activity. These findings indicate that PARP-1 plays a pivotal role in the initial inflammatory response by modulating transcription of inflammation-linked genes. PMID- 11854474 TI - CFTR with a partially deleted R domain corrects the cystic fibrosis chloride transport defect in human airway epithelia in vitro and in mouse nasal mucosa in vivo. AB - In developing gene therapy for cystic fibrosis (CF) airways disease, a transgene encoding a partially deleted CF transmembrane conductance regulator (CFTR) Cl- channel could be of value for vectors such as adeno-associated virus that have a limited packaging capacity. Earlier studies in heterologous cells indicated that the CFTR R (regulatory) domain is predominantly random coil and that parts of the R domain can be deleted without abolishing channel function. Therefore, we designed a series of CFTR variants with shortened R domains (between residues 708 and 835) and expressed them in well-differentiated cultures of CF airway epithelia. All of the variants showed normal targeting to the apical membrane, and for the constructs we tested, biosynthesis was like wild type. Moreover, all constructs generated transepithelial Cl- current in CF epithelia. Comparison of the Cl- transport suggested that the length of the R domain, the presence of phosphorylation sites, and other factors contribute to channel activity. A variant deleting residues 708-759 complemented CF airway epithelia to the same extent as wild-type CFTR and showed no current in the absence of cAMP stimulation. In addition, expression in nasal mucosa of CF mice corrected the Cl- transport defect. These data provide insight into the structure and function of the R domain and identify regions that can be deleted with retention of function. Thus they suggest a strategy for shortening the transgene used in CF gene therapy. PMID- 11854475 TI - Mortality in 7B2 null mice can be rescued by adrenalectomy: involvement of dopamine in ACTH hypersecretion. AB - The serine protease prohormone convertase 2 (PC2), principally involved in the processing of polypeptide hormone precursors in neuroendocrine tissues, requires interaction with the neuroendocrine protein 7B2 to generate an enzymatically active form. 7B2 null mice express no PC2 activity and release large quantities of uncleaved ACTH, resulting in a lethal endocrine condition that resembles pituitary Cushing's (Westphal, C. H., Muller, L., Zhou, A., Bonner-Weir, S., Schambelan, M., Steiner, D. F., Lindberg, I. & Leder, P. (1999) Cell 96, 689). Here, we have compared the 7B2 and PC2 null mouse models to determine why the 7B2 null, but not the PC2 null, exhibits a lethal disease state. Both 7B2 and PC2 nulls contained highly elevated pituitary adrenocorticotropic hormone (ACTH); the neurointermediate lobe content of ACTH in 7B2 nulls was 13-fold higher than in WT mice; that of the PC2 null was 65-fold higher. However, circulating ACTH levels were much higher in the 7B2 null than in the PC2 null. Because hypothalamic inhibitory dopaminergic control represents the major influence on intermediate lobe proopiomelanocortin-derived peptide secretion, dopamine levels were measured, and they revealed that 7B2 null pituitaries contained only one-fourth of WT pituitary dopamine. Adrenalectomized 7B2 null animals survived past the usual time of death at 5 weeks; a month after adrenalectomy, they exhibited normal levels of pituitary dopamine, circulating ACTH, and corticosterone. Elevated corticosterone, therefore, seems to play a central role in the lethal phenotype of the 7B2 null, whereas a 7B2-mediated dopaminergic deficiency state may be involved in the actual ACTH hypersecretion phenomenon. Interestingly, adrenalectomized 7B2 nulls also developed unexpectedly severe obesity. PMID- 11854476 TI - HIV's response to a CCR5 inhibitor: I'd rather tighten than switch! PMID- 11854477 TI - New insights into unwrapping DNA from the nucleosome from a single-molecule optical tweezers method. PMID- 11854479 TI - The role of intersection topography in bond selectivity of cis-trans photoisomerization. AB - Ab initio methods are used to characterize the ground and first excited state of the chromophore in the rhodopsin family of proteins: retinal protonated Schiff base. Retinal protonated Schiff base has five double bonds capable of undergoing isomerization. Upon absorption of light, the chromophore isomerizes and the character of the photoproducts (e.g., 13-cis and 11-cis) depends on the environment, protein vs. solution. Our ab initio calculations show that, in the absence of any specific interactions with the environment (e.g., discrete ordered charges in a protein), energetic considerations cannot explain the observed bond selectivity. We instead attribute the origin of bond selectivity to the shape (topography) of the potential energy surfaces in the vicinity of points of true degeneracy (conical intersections) between the ground and first excited electronic states. This provides a molecular example where a competition between two distinct but nearly isoenergetic photochemical reaction pathways is resolved by a topographical difference between two conical intersections. PMID- 11854478 TI - Will embryonic stem cells be a useful source of dopamine neurons for transplant into patients with Parkinson's disease? PMID- 11854481 TI - Influence of anthropogenic aerosol on cloud optical depth and albedo shown by satellite measurements and chemical transport modeling. AB - The Twomey effect of enhanced cloud droplet concentration, optical depth, and albedo caused by anthropogenic aerosols is thought to contribute substantially to radiative forcing of climate change over the industrial period. However, present model-based estimates of this indirect forcing are highly uncertain. Satellite based measurements would provide global or near-global coverage of this effect, but previous efforts to identify and quantify enhancement of cloud albedo caused by anthropogenic aerosols in satellite observations have been limited, largely because of strong dependence of albedo on cloud liquid water path (LWP), which is inherently highly variable. Here we examine satellite-derived cloud radiative properties over two 1-week episodes for which a chemical transport and transformation model indicates substantial influx of sulfate aerosol from industrial regions of Europe or North America to remote areas of the North Atlantic. Despite absence of discernible dependence of optical depth or albedo on modeled sulfate loading, examination of the dependence of these quantities on LWP readily permits detection and quantification of increases correlated with sulfate loading, which are otherwise masked by variability of LWP, demonstrating brightening of clouds because of the Twomey effect on a synoptic scale. Median cloud-top spherical albedo was enhanced over these episodes, relative to the unperturbed base case for the same LWP distribution, by 0.02 to 0.15. PMID- 11854482 TI - Optimizing bioconversion pathways through systems analysis and metabolic engineering. AB - We demonstrate a general approach for metabolic engineering of biocatalytic systems comprising the uses of a chemostat for strain improvement and radioisotopic tracers for the quantification of pathway fluxes. Flux determination allows the identification of target pathways for modification as validated by subsequent overexpression of the corresponding gene. We demonstrate this method in the indene bioconversion network of Rhodococcus modified for the overproduction of 1,2-indandiol, a key precursor for the AIDS drug Crixivan. PMID- 11854483 TI - Retrocyclin: a primate peptide that protects cells from infection by T- and M tropic strains of HIV-1. AB - Human bone marrow expresses a pseudogene that encodes an antimicrobial peptide homologous to rhesus monkey circular minidefensins (delta-defensins). We prepared the putative ancestral human peptide by solid-phase synthesis and named it "retrocyclin." Retrocyclin did not cause direct inactivation of HIV-1, and its modest antibacterial properties resembled those of its rhesus homologs. Nevertheless, retrocyclin had a remarkable ability to inhibit proviral DNA formation and to protect immortalized and primary human CD4(+) lymphocytes from in vitro infection by both T-tropic and M-tropic strains of HIV-1. Confocal fluorescent microscopy studies performed with BODIPY-FL-labeled RC-101, a close analog of retrocyclin, showed that the peptide formed patch-like aggregates on the surface of CD4(+) cells. These findings suggest that retrocyclin interferes with an early stage of HIV-1 infection and that retrocyclin-like agents might be useful topical agents to prevent sexually acquired HIV-1 infections. PMID- 11854484 TI - Polypeptide release at sense and noncognate stop codons by localized charge exchange alterations in translational release factors. AB - The mechanism of stop codon recognition during translation has long been a puzzle. Only recently has it been established that a tripeptide in the bacterial release factors (RFs) 1 and 2 serves as the "anticodon" in deciphering stop codons in mRNA. However, the molecular basis of the accuracy of stop codon recognition is unknown. Although specific tripeptides in the RFs are primarily responsible for selective reading of cognate stop codons, charge-flip variant RF proteins, altered at conserved Glu residues adjacent to the tripeptide-anticodon, are shown here to be crucial to codon recognition. Changes of these Glu residues are capable of triggering polypeptide release at noncognate stop codons and also at sense codons. These changes also reverse the growth inhibition by RFs containing "harmful" tripeptide-anticodon changes. These findings suggest that electrostatic interactions involving negative charges in domain C of the RFs mediate their accurate docking in the ribosome. Our results also establish that the charge flipping creates a phenotype/translation termination by "codon bypassing" via relaxed positioning of the RF tripeptide-anticodon in the decoding pocket of the ribosome. PMID- 11854486 TI - Energetics by NMR: site-specific binding in a positively cooperative system. AB - Proteins with multiple binding sites exhibit a complex behavior that depends on the intrinsic affinities for each site and the energetic communication between the sites. The contributions from intrinsic affinity and cooperativity are difficult to deconvolute using conventional binding experiments that lack information about the occupancies of individual sites. Here, we report the concerted use of NMR and isothermal titration calorimetry to determine the intrinsic and cooperative binding free energies for a ligand-protein complex. The NMR measurements provided the site-specific information necessary to resolve the binding parameters. Using this approach, we observed that human ileal bile acid binding protein binds two molecules of glycocholic acid with low intrinsic affinity but an extraordinarily high degree of positive cooperativity. The highly cooperative nature of the binding provides insights into the protein's biological mechanism. With ongoing improvements in sensitivity and resolution, NMR methods are becoming more amenable to dissecting the complex binding energetics of multisite systems. PMID- 11854485 TI - An NMR approach to structural proteomics. AB - The influx of genomic sequence information has led to the concept of structural proteomics, the determination of protein structures on a genome-wide scale. Here we describe an approach to structural proteomics of small proteins using NMR spectroscopy. Over 500 small proteins from several organisms were cloned, expressed, purified, and evaluated by NMR. Although there was variability among proteomes, overall 20% of these proteins were found to be readily amenable to NMR structure determination. NMR sample preparation was centralized in one facility, and a distributive approach was used for NMR data collection and analysis. Twelve structures are reported here as part of this approach, which allowed us to infer putative functions for several conserved hypothetical proteins. PMID- 11854487 TI - Feeding acetyl-L-carnitine and lipoic acid to old rats significantly improves metabolic function while decreasing oxidative stress. AB - Mitochondrial-supported bioenergetics decline and oxidative stress increases during aging. To address whether the dietary addition of acetyl-l-carnitine [ALCAR, 1.5% (wt/vol) in the drinking water] and/or (R)-alpha-lipoic acid [LA, 0.5% (wt/wt) in the chow] improved these endpoints, young (2-4 mo) and old (24-28 mo) F344 rats were supplemented for up to 1 mo before death and hepatocyte isolation. ALCAR+LA partially reversed the age-related decline in average mitochondrial membrane potential and significantly increased (P = 0.02) hepatocellular O(2) consumption, indicating that mitochondrial-supported cellular metabolism was markedly improved by this feeding regimen. ALCAR+LA also increased ambulatory activity in both young and old rats; moreover, the improvement was significantly greater (P = 0.03) in old versus young animals and also greater when compared with old rats fed ALCAR or LA alone. To determine whether ALCAR+LA also affected indices of oxidative stress, ascorbic acid and markers of lipid peroxidation (malondialdehyde) were monitored. The hepatocellular ascorbate level markedly declined with age (P = 0.003) but was restored to the level seen in young rats when ALCAR+LA was given. The level of malondialdehyde, which was significantly higher (P = 0.0001) in old versus young rats, also declined after ALCAR+LA supplementation and was not significantly different from that of young unsupplemented rats. Feeding ALCAR in combination with LA increased metabolism and lowered oxidative stress more than either compound alone. PMID- 11854489 TI - Forced engagement of a RNA/protein complex by a chemical inducer of dimerization to modulate gene expression. AB - A general strategy is described for forcing the engagement of an RNA/protein complex by using small-molecule ligands. A bivalent molecule was created by linking a protein-binding ligand to an RNA-binding ligand. On presentation of the chemical inducer of dimerization to the RNA by the protein, cooperative binding ensued, resulting in higher-affinity complexes. When the chemical inducer of dimerization was used to target the protein to an mRNA template, the resulting RNA/protein complex was sufficiently stable to inhibit mRNA translation. This approach provides a logic to modulate gene expression by using small-molecule ligands to recruit protein surfaces to mRNAs. PMID- 11854488 TI - Age-associated mitochondrial oxidative decay: improvement of carnitine acetyltransferase substrate-binding affinity and activity in brain by feeding old rats acetyl-L- carnitine and/or R-alpha -lipoic acid. AB - We test whether the dysfunction with age of carnitine acetyltransferase (CAT), a key mitochondrial enzyme for fuel utilization, is due to decreased binding affinity for substrate and whether this substrate, fed to old rats, restores CAT activity. The kinetics of CAT were analyzed by using the brains of young and old rats and of old rats supplemented for 7 weeks with the CAT substrate acetyl-l carnitine (ALCAR) and/or the mitochondrial antioxidant precursor R-alpha-lipoic acid (LA). Old rats, compared with young rats, showed a decrease in CAT activity and in CAT-binding affinity for both substrates, ALCAR and CoA. Feeding ALCAR or ALCAR plus LA to old rats significantly restored CAT-binding affinity for ALCAR and CoA, and CAT activity. To explore the underlying mechanism, lipid peroxidation and total iron and copper levels were assayed; all increased in old rats. Feeding old rats LA or LA plus ALCAR inhibited lipid peroxidation but did not decrease iron and copper levels. Ex vivo oxidation of young-rat brain with Fe(II) caused loss of CAT activity and binding affinity. In vitro oxidation of purified CAT with Fe(II) inactivated the enzyme but did not alter binding affinity. However, in vitro treatment of CAT with the lipid peroxidation products malondialdehyde or 4-hydroxy-nonenal caused a decrease in CAT-binding affinity and activity, thus mimicking age-related change. Preincubation of CAT with ALCAR or CoA prevented malondialdehyde-induced dysfunction. Thus, feeding old rats high levels of key mitochondrial metabolites can ameliorate oxidative damage, enzyme activity, substrate-binding affinity, and mitochondrial dysfunction. PMID- 11854490 TI - The crystal structure and mutational analysis of a novel RNA-binding domain found in the human Tap nuclear mRNA export factor. AB - The Tap protein mediates the sequence nonspecific nuclear export of cellular mRNAs as well as the sequence-specific export of retroviral mRNAs bearing the constitutive transport element (CTE). Previously, the structures of individual Tap subdomains, including ribonucleoprotein and leucine-rich repeat domains, have been described. Here, we report the crystal structure of a functional CTE RNA binding domain of human Tap, including the N-terminal arm of the ribonucleoprotein domain and interdomain linking polypeptide. To identify residues that interact with the CTE, we have introduced 38 alanine substitutions for surface residues in the Tap CTE-binding domain and tested these mutants for their ability to support CTE-dependent nuclear RNA export and CTE binding. Four residues that cluster on a concave surface in the leucine-rich repeat domain were found to be critical for CTE binding and define a CTE-interacting surface on this domain. The second critical CTE-interacting surface on Tap is defined by three previously identified residues on the surface of the ribonucleoprotein domain. The structural and mutational data define a novel RNA-binding site on the Tap protein. PMID- 11854492 TI - Differential effects of a centrally acting fatty acid synthase inhibitor in lean and obese mice. AB - C75 is a potent inhibitor of fatty acid synthase that acts centrally to reduce food intake and body weight in mice; a single dose causes a rapid (>90%) decrease of food intake. These effects are associated with inhibition of fasting-induced up-regulation and down-regulation, respectively, of the expression of orexigenic (NPY and AgRP) and anorexigenic (POMC and CART) neuropeptide messages in the hypothalamus. Repeated administration of C75 at a submaximal level, however, differentially affected food intake of lean and obese mice. With lean mice, C75 suppressed food intake by approximately 50% and, with obese mice (ob/ob and dietary-induced obesity), by 85-95% during the first day of treatment. Lean mice, however, became tolerant/resistant to C75 over the next 2-5 days of treatment, with food intake returning to near normal and rebound hyperphagia occurring on cessation of treatment. In contrast, ob/ob obese mice responded to C75 with a >90% suppression of food intake throughout the same period with incipient tolerance becoming evident only after substantial weight loss had occurred. Dietary-induced obese mice exhibited intermediate behavior. In all cases, a substantial loss of body weight resulted. Pair-fed controls lost 24-50% less body weight than C75-treated mice, indicating that, in addition to suppressing food intake, C75 may increase energy expenditure. The decrease in body weight by ob/ob mice was due primarily to loss of body fat. In contrast to the short-term effects of C75 on "fasting-induced" changes of hypothalamic orexigenic and anorexigenic neuropeptide mRNAs, repeated administration of C75 either had the inverse or no effect as tolerance developed. PMID- 11854491 TI - The human DnaJ protein, hTid-1, enhances binding of a multimer of the herpes simplex virus type 1 UL9 protein to oris, an origin of viral DNA replication. AB - We have identified cellular proteins that interact with the herpes simplex virus type 1 (HSV-1) origin-binding protein (UL9 protein) by screening a HeLa cell complementary DNA library by using the yeast two-hybrid system. Approximately 7 x 10(5) colonies were screened. Five of the 48 positive clones contained cDNAs that encoded the p150(Glued) component of the dynactin complex, three contained cDNAs for the neural F Box 42-kDa protein (NFB42), which is highly enriched in neural tissue, and three contained hTid-1, a human homologue of the bacterial DnaJ protein. We have focused in this report on the interaction of the viral UL9 protein with the cellular hTid-1. In vitro immunoprecipitation experiments confirmed that hTid-1 interacts with the UL9 protein. Electrophoretic mobility shift assays indicated that the hTid-1 enhances the binding of UL9 protein to an HSV-1 origin, ori(s), and facilitates formation of the multimer from the dimeric UL9 protein. hTid-1 had no effect on the DNA-dependent ATPase or helicase activities associated with the UL9 protein. These findings implicate hTid-1 in HSV-1 DNA replication, and suggest that this cellular protein may provide a chaperone function analogous to the DnaJ protein in Escherichia coli DNA replication. PMID- 11854493 TI - The carboxyl tail forms a discrete functional domain that blocks closure of the yeast K+ channel. AB - Non-targeted mutagenesis studies of the yeast K(+) channel, TOK1, have led to identification of functional domains common to other cation channels as well as those so far not found in other channels. Among the latter is the ability of the carboxyl tail to prevent channel closure. Here, we show that the tail can fulfill this function in trans. Coexpression of the carboxyl tail with the tail-deleted channel core restores normal channel behavior A Ser/Thr-rich region at its amino end and an acidic stretch at its carboxyl end delineate the minimal region required for tail function. This region of 160 aa apparently forms a discrete functional domain. Interaction of this domain with the channel core is strong, being recalcitrant to removal from excised membrane patches by both high salt and reducing agents. Although the use of a cytoplasmic domain to regulate channel is common among animal channels, by using it as a "foot-in-the-door" to maintain open state appears unique to TOK1, the first fungal K(+) channel studied in depth. PMID- 11854494 TI - A method for optimizing potential-energy functions by a hierarchical design of the potential-energy landscape: application to the UNRES force field. AB - A method for optimizing potential-energy functions of proteins is proposed. The method assumes a hierarchical structure of the energy landscape, which means that the energy decreases as the number of native-like elements in a structure increases, being lowest for structures from the native family and highest for structures with no native-like element. A level of the hierarchy is defined as a family of structures with the same number of native-like elements (or degree of native likeness). Optimization of a potential-energy function is aimed at achieving such a hierarchical structure of the energy landscape by forcing appropriate free-energy gaps between hierarchy levels to place their energies in ascending order. This procedure is different from methods developed thus far, in which the energy gap and/or the Z score between the native structure and all non native structures are maximized, regardless of the degree of native likeness of the non-native structures. The advantage of this approach lies in reducing the number of structures with decreasing energy, which should ensure the searchability of the potential. The method was tested on two proteins, PDB ID codes and, with an off-lattice united-residue force field. For, the search of the conformational space with the use of the conformational space annealing method and the newly optimized potential-energy function found the native structure very quickly, as opposed to the potential-energy functions obtained by former optimization methods. After even incomplete optimization, the force field obtained by using located the native-like structures of two peptides, and betanova (a designed three-stranded beta-sheet peptide), as the lowest-energy conformations, whereas for the 46-residue N-terminal fragment of staphylococcal protein A, the native-like conformation was the second-lowest-energy conformation and had an energy 2 kcal/mol above that of the lowest-energy structure. PMID- 11854495 TI - Mechanical disruption of individual nucleosomes reveals a reversible multistage release of DNA. AB - The dynamic structure of individual nucleosomes was examined by stretching nucleosomal arrays with a feedback-enhanced optical trap. Forced disassembly of each nucleosome occurred in three stages. Analysis of the data using a simple worm-like chain model yields 76 bp of DNA released from the histone core at low stretching force. Subsequently, 80 bp are released at higher forces in two stages: full extension of DNA with histones bound, followed by detachment of histones. When arrays were relaxed before the dissociated state was reached, nucleosomes were able to reassemble and to repeat the disassembly process. The kinetic parameters for nucleosome disassembly also have been determined. PMID- 11854496 TI - Functional studies on the role of the C-terminal domain of mammalian polo-like kinase. AB - Mammalian polo-like kinase (Plk) acts at various stages in early and late mitosis. Plk is phosphorylated and activated in mitosis, and the proper subcellular localization of Plk is essential for mitotic regulation. We have observed that overexpression of the C-terminal domain of Plk is more effective than wild-type or kinase-defective Plk in causing mitotic delay or arrest. The specific activity of Plk with C-terminal deletions or substitution of aspartate for threonine-210 is increased severalfold relative to wild type. We show in this communication that the C-terminal domain can bind to full-length or the catalytic domain of Plk and inhibit its kinase activity, and that this binding is disrupted when threonine-210 is substituted with an aspartic acid residue. The C-terminal domain binds unphosphorylated Plk from G(2) arrested cells, but not phosphorylated Plk from mitotic cells. Green fluorescent protein-C-terminal Plk is localized at the centrosome and the midbody of transfected cells as shown previously for full-length enzyme. These and other data indicate that although the C terminus serves to regulate Plk kinase activity, the localization of the C terminus at the centrosome and other sites in transfected cells may block the correct localization of endogenous Plk. PMID- 11854497 TI - Targeting endothelium and its dynamic caveolae for tissue-specific transcytosis in vivo: a pathway to overcome cell barriers to drug and gene delivery. AB - Site-directed pharmacodelivery is a desirable but elusive goal. Endothelium and epithelium create formidable barriers to endogenous molecules as well as targeted therapies in vivo. Caveolae provide a possible, yet unproven, transcellular pathway to overcome such barriers. By using an antibody- and subfractionation based strategy, we generated a monoclonal antibody specific for lung caveolae (TX3.833) that targets rat lungs after i.v. injection (up to 89% of dose in 30 min). Unlike control antibodies (nonbinding or to lipid rafts), TX3.833 targets lung caveolae that bud to form free vesicles for selective and quantal transendothelial transport to underlying tissue cells in vivo. Rapid sequential transcytosis can occur to the alveolar air space via epithelial caveolae. Conjugation to TX3.833 increases drug delivery to the lung up to 172-fold and achieves rapid, localized bioefficacy. We conclude that: (i) molecular heterogeneity of the endothelium and its caveolae permits vascular targeting to achieve theoretical expectations of tissue-specific delivery and bioefficacy; (ii) caveolae can mediate selective transcytosis in vivo; and (iii) targeting caveolae may provide a tissue-specific pathway for overcoming key cell barriers to many drug and gene therapies in vivo. PMID- 11854498 TI - Mutagenesis of a functional chimeric gene in yeast identifies mutations in the simian virus 40 large T antigen J domain. AB - Simian virus 40 large T antigen contains an amino terminal J domain that catalyzes T antigen-mediated viral DNA replication and cellular transformation. To dissect the role of the J domain in these processes, we exploited the genetic tools available only in the yeast Saccharomyces cerevisiae to isolate 14 loss-of function point mutations in the T antigen J domain. This screen also identified mutations that, when engineered into simian virus 40, resulted in T antigen mutants that were defective for the ability to support viral growth, to transform mammalian cells in culture, to dissociate the p130-E2F4 transcription factor complex, and to stimulate ATP hydrolysis by hsc70, a hallmark of J domain containing molecular chaperones. These data correlate the chaperone activity of the T antigen J domain with its roles in viral infection and cellular transformation and support a model by which the viral J domain recruits the cytoplasmic hsc70 molecular chaperone in the host to rearrange multiprotein complexes implicated in replication and transformation. More generally, this study presents the use of a yeast screen to identify loss-of-function mutations in a mammalian virus and can serve as a widely applicable method to uncover domain functions of mammalian proteins for which there are yeast homologues with selectable mutant phenotypes. PMID- 11854500 TI - Suppression of proliferation and cardiomyocyte hypertrophy by CHAMP, a cardiac specific RNA helicase. AB - Adult cardiomyocytes are irreversibly postmitotic but respond to a variety of stimuli by hypertrophic growth, which is associated with an increase in cell size and protein content, organization of sarcomeres, and activation of a fetal gene program. Recently, we described a novel cardiac helicase activated by MEF2 protein (CHAMP), which is expressed specifically in the heart throughout prenatal and postnatal development. Here we show that CHAMP acts as an inhibitor of cell proliferation and cardiomyocyte hypertrophy. Ectopic expression of CHAMP inhibits proliferation of HeLa cells and blocks cell cycle entry of serum-stimulated NIH 3T3 cells. Overexpression of CHAMP in primary neonatal cardiomyocytes blocks hypertrophic growth and the induction of fetal genes in response to stimulation by serum and phenylephrine but does not prevent sarcomere organization or early mitogenic signaling events including activation of extracellular signal-regulated kinases or up-regulation of c-fos. Inhibition of cardiomyocyte hypertrophy by CHAMP requires the conserved ATPase domain and is accompanied by up-regulation of the cyclin-dependent protein kinase inhibitor p21(CIP1). These findings identify CHAMP as a cardiac-specific suppressor of cardiomyocyte hypertrophy and cell cycle progression and suggest that CHAMP may suppress these processes through the regulation of p21(CIP1). PMID- 11854501 TI - Impact of the terminal Cretaceous event on plant-insect associations. AB - Evidence for a major extinction of insect herbivores is provided by presence absence data for 51 plant-insect associations on 13,441 fossil plant specimens, spanning the Cretaceous/Paleogene boundary in southwestern North Dakota. The most specialized associations, which were diverse and abundant during the latest Cretaceous, almost disappeared at the boundary and failed to recover in younger strata even while generalized associations regained their Cretaceous abundances. These results are consistent with a sudden ecological perturbation that precipitated a diversity bottleneck for insects and plants. PMID- 11854499 TI - p56Lck anchors CD4 to distinct microdomains on microvilli. AB - Cell-surface microvilli play a central role in adhesion, fusion, and signaling processes. Some adhesion and signaling receptors segregate on microvilli but the determinants of this localization remain mostly unknown. In this study, we considered CD4, a receptor involved in immune response and HIV infection, and p56(Lck), a CD4-associated tyrosine kinase. Analysis of CD4 trafficking reveals that p56(Lck) binds tightly to CD4 independently of its activation state and inhibits CD4 internalization. Electron microscopy analysis established that p56(Lck) mediates CD4 association with microvilli whereas biochemical data indicate that p56(Lck) expression renders CD4 insoluble by the nonionic detergent Triton X-100. In addition, cytoskeleton-disrupting agent increased CD4 solubility, suggesting the involvement of cytoskeletal elements in CD4 anchoring to microvilli. This concept was supported further by the observation that the lateral mobility of CD4 within the plasma membrane was decreased in cells expressing p56(Lck). Finally, isolation of detergent-resistant membranes revealed that the complex CD4-p56(Lck) is enriched within these domains as opposed to conditions in which CD4 does not interact with p56(Lck). In conclusion, our results show that p56(Lck) targets CD4 to specialized lipid microdomains preferentially localized on microvilli. This localization, which prevents CD4 internalization, might facilitate CD4-mediated adhesion processes and could correspond to the signaling site of the receptor. PMID- 11854502 TI - Hotspots and the conservation of evolutionary history. AB - Species diversity is unevenly distributed across the globe, with terrestrial diversity concentrated in a few restricted biodiversity hotspots. These areas are associated with high losses of primary vegetation and increased human population density, resulting in growing numbers of threatened species. We show that conservation of these hotspots is critical because they harbor even greater amounts of evolutionary history than expected by species numbers alone. We used supertrees for carnivores and primates to estimate that nearly 70% of the total amount of evolutionary history represented in these groups is found in 25 biodiversity hotspots. PMID- 11854503 TI - Specificity and robustness in transcription control networks. AB - Recognition by transcription factors of the regulatory DNA elements upstream of genes is the fundamental step in controlling gene expression. How does the necessity to provide stability with respect to mutation constrain the organization of transcription control networks? We examine the mutation load of a transcription factor interacting with a set of n regulatory response elements as a function of the factor/DNA binding specificity and conclude on theoretical grounds that the optimal specificity decreases with n. The predicted correlation between variability of binding sites (for a given transcription factor) and their number is supported by the genomic data for Escherichia coli. The analysis of E. coli genomic data was carried out using an algorithm suggested by the biophysical model of transcription factor/DNA binding. Complete results of the search for candidate transcription factor binding sites are available at http://www.physics.rockefeller.edu/~boris/public/search_ecoli. PMID- 11854504 TI - Integration of foreign DNA during natural transformation of Acinetobacter sp. by homology-facilitated illegitimate recombination. AB - The active uptake of extracellular DNA and its genomic integration is termed natural transformation and constitutes a major horizontal gene-transfer mechanism in prokaryotes. Chromosomal DNA transferred within a species can be integrated effectively by homologous recombination, whereas foreign DNA with low or no sequence homology would rely on illegitimate recombination events, which are rare. By using the nptII(+) gene (kanamycin resistance) as selectable marker, we found that the integration of foreign DNA into the genome of the Gram-negative Acinetobacter sp. BD413 during transformation indeed was at least 10(9)-fold lower than that of homologous DNA. However, integration of foreign DNA increased at least 10(5)-fold when it was linked on one side to a piece of DNA homologous to the recipient genome. Analysis of foreign DNA integration sites revealed short stretches of sequence identity (3-8 bp) between donor and recipient DNA, indicating illegitimate recombination events. These findings suggest that homologous DNA served as a recombinational anchor facilitating illegitimate recombination acting on the same molecule. Homologous stretches down to 183 nucleotides served as anchors. Transformation with heteroduplex DNA having different nucleotide sequence tags in the strands indicated that strands entered the cytoplasm 3' to 5' and that strands with either polarity were integrated by homologous recombination. The process led to the genomic integration of thousands of foreign nucleotides and often was accompanied by deletion of a roughly corresponding length of recipient DNA. Homology-facilitated illegitimate recombination would explain the introgression of DNA in prokaryotic genomes without the help of mobile genetic elements. PMID- 11854505 TI - Homologous recombination at the border: insertion-deletions and the trapping of foreign DNA in Streptococcus pneumoniae. AB - Integration of foreign DNA was observed in the Gram-positive human pathogen Streptococcus pneumoniae (pneumococcus) after transformation with DNA from a recombinant Escherichia coli bacteriophage lamda carrying a pneumococcal insert. Segments of lamda DNA replaced chromosomal sequences adjacent to the region homologous with the pneumococcal insert, whence the name insertion-deletion. Here we report that a pneumococcal insert was absolutely required for insertion deletion formation, but could be as short as 153 bp; that the sizes of foreign DNA insertions (289-2,474 bp) and concomitant chromosomal deletions (45-1,485 bp) were not obviously correlated; that novel joints clustered preferentially within segments of high GC content; and that the crossovers in 29 independent novel joints were located 1 bp from the border or within short (3-10 nt long) stretches of identity (microhomology) between resident and foreign DNA. The data are consistent with a model in which the insert serving as a homologous recombination anchor favors interaction and subsequent illegitimate recombination events at microhomologies between foreign and resident sequences. The potential of homology directed illegitimate recombination for genome evolution was illustrated by the trapping of functional heterologous genes. PMID- 11854506 TI - A comparative radiation hybrid map of bovine chromosome 18 and homologous chromosomes in human and mice. AB - A comprehensive radiation hybrid (RH) map and a high resolution comparative map of Bos taurus (BTA) chromosome 18 were constructed, composed of 103 markers and 76 markers, respectively, by using a cattle-hamster somatic hybrid cell panel and a 5,000 rad whole-genome radiation hybrid (WGRH) panel. These maps include 65 new assignments (56 genes, 3 expressed-sequence tags, 6 microsatellites) and integrate 38 markers from the first generation WGRH(5,000) map of BTA18. Fifty nine assignments of coding sequences were supported by somatic hybrid cell mapping to markers on BTA18. The total length of the comprehensive map was 1666 cR(5,000). Break-point positions within the chromosome were refined and a new telomeric RH linkage group was established. Conserved synteny between cattle, human, and mouse was found for 76 genes of BTA18 and human chromosomes (HSA) 16 and 19 and for 34 cattle genes and mouse chromosomes (MMU) 7 and 8. The new RH map is potentially useful for the identification of candidate genes for economically important traits, contributes to the expansion of the existing BTA18 gene map, and provides new information about the chromosome evolution in cattle, humans, and mice. PMID- 11854507 TI - Analysis of DNA microarrays using algorithms that employ rule-based expert knowledge. AB - The ability to investigate the transcription of thousands of genes concurrently by using DNA microarrays offers both major scientific opportunities and significant analytical challenges. Here we describe GABRIEL, a rule-based system of computer programs designed to apply domain-specific and procedural knowledge systematically and uniformly for the analysis and interpretation of data from DNA microarrays. GABRIEL'S problem-solving rules direct stereotypical tasks, whereas domain-specific knowledge pertains to gene functions and relationships or to experimental conditions. Additionally, GABRIEL can learn novel rules through genetic algorithms, which define patterns that best match the data being analyzed and can identify groupings in gene expression profiles preordered by chromosomal position or by a nonsupervised algorithm such as hierarchical clustering. GABRIEL subsystems explain the logic that underlies conclusions and provide a graphical interface and interactive platform for the acquisition of new knowledge. The present report compares GABRIEL'S output with published findings in which expert knowledge has been applied post hoc to microarray groupings generated by hierarchical clustering. PMID- 11854508 TI - Discovery of five conserved beta -defensin gene clusters using a computational search strategy. AB - The innate immune system includes antimicrobial peptides that protect multicellular organisms from a diverse spectrum of microorganisms. beta-Defensins comprise one important family of mammalian antimicrobial peptides. The annotation of the human genome fails to reveal the expected diversity, and a recent query of the draft sequence with the blast search engine found only one new beta-defensin gene (DEFB3). To define better the beta-defensin gene family, we adopted a genomics approach that uses hmmer, a computational search tool based on hidden Markov models, in combination with blast. This strategy identified 28 new human and 43 new mouse beta-defensin genes in five syntenic chromosomal regions. Within each syntenic cluster, the gene sequences and organization were similar, suggesting each cluster pair arose from a common ancestor and was retained because of conserved functions. Preliminary analysis indicates that at least 26 of the predicted genes are transcribed. These results demonstrate the value of a genomewide search strategy to identify genes with conserved structural motifs. Discovery of these genes represents a new starting point for exploring the role of beta-defensins in innate immunity. PMID- 11854510 TI - Transgenesis by lentiviral vectors: lack of gene silencing in mammalian embryonic stem cells and preimplantation embryos. AB - The introduction of foreign genes into early mouse embryos and embryonic stem (ES) cells is invaluable for the analysis of gene function and regulation in the living animal. The use of vectors derived from retroviruses as gene transfer vehicles in this setting has had limited success because of silencing of transgene expression. Here, we show that vectors derived from lentiviruses, which are complex retroviruses, can efficiently deliver genes to murine ES cells and that transgene expression is stable during proliferation of undifferentiated ES cells. The transgene is expressed during differentiation of ES cells in vitro (embryoid bodies) and in vivo (teratomas). Transfer of lentivector-transduced ES cells into blastocysts resulted in chimeric animals that expressed the transgene in multiple tissues. Embryos derived from crossings of chimeric mice expressed the transgene, indicating successful germ-line transmission. Infection of murine preimplantation embryos at morula stage with lentiviral vectors resulted in stable transduction and expression of the transgene in mouse embryos and in newborn mice. Finally, human ES cells were transduced by lentiviral vectors and expressed the transgene over several passages. Thus, lentiviral vectors represent a significant improvement over oncoretroviral vectors used previously for gene transfer into murine ES cells and preimplantation embryos. Ability to transfer foreign genes into human ES cells has potential relevance for the development of gene and cell-based therapies. PMID- 11854511 TI - T-DNA integration into the barley genome from single and double cassette vectors. AB - Patterns and sites of T-DNA integrations into the barley genome from single and double cassette vectors are of interest for the identification of cultivars with value added properties as well as for the production of selection marker-free transgenic lines that can be retransformed. T-DNA/Plant DNA junctions were obtained by capturing a single-stranded DNA with a biotinylated primer annealing to the vector adjacent to the border and an adaptor ligated to a restriction site overhang in the flanking barley DNA. The captured junction was converted into a double strand and sequenced. Fifty left and right border junctions from plants transgenic for one of five human genes were analyzed. Primers of 15-30 nucleotides designed from the genomic DNA at the insertion site can PCR amplify fragments that identify unequivocally any transformant. Adjacent transgene insertions with single cassette vectors were always in tandem direct repeat configuration. With regard to T-DNA integration the patterns were comparable to the variations found in dicotyledonous plants. Twelve of the 46 integrations characterized by blast searches were within different regions of the BARE-1 retrotransposon element occurring with a frequency of 2 x 10(5) copies in the barley genome. The use of border junctions to identify number of copies and loci of integrates in transformants is discussed. PMID- 11854509 TI - Loss of circadian clock function decreases reproductive fitness in males of Drosophila melanogaster. AB - Circadian coordination of life functions is believed to contribute to an organism's fitness; however, such contributions have not been convincingly demonstrated in any animal. The most significant measure of fitness is the reproductive output of the individual and species. Here we examined the consequences of loss of clock function on reproductive fitness in Drosophila melanogaster with mutated period (per(0)), timeless (tim(0)), cycle (cyc(0)), and Clock (Clk(Jrk)) genes. Single mating among couples with clock-deficient phenotypes resulted in approximately 40% fewer progeny compared with wild-type flies, because of a decreased number of eggs laid and a greater rate of unfertilized eggs. Male contribution to this phenotype was demonstrated by a decrease in reproductive capacity among per(0) and tim(0) males mated with wild type females. The important role of clock genes for reproductive fitness was confirmed by reversal of the low-fertility phenotype in flies with rescued per or tim function. Males lacking a functional clock showed a significant decline in the quantity of sperm released from the testes to seminal vesicles, and these tissues displayed rhythmic and autonomous expression of clock genes. By combining molecular and physiological approaches, we identified a circadian clock in the reproductive system and defined its role in the sperm release that promotes reproductive fitness in D. melanogaster. PMID- 11854512 TI - Constitutive expression of a single antimicrobial peptide can restore wild-type resistance to infection in immunodeficient Drosophila mutants. AB - One of the characteristics of the host defense of insects is the rapid synthesis of a variety of potent antibacterial and antifungal peptides. To date, seven types of inducible antimicrobial peptides (AMPs) have been characterized in Drosophila. The importance of these peptides in host defense is supported by the observation that flies deficient for the Toll or Immune deficiency (Imd) pathway, which affects AMP gene expression, are extremely susceptible to microbial infection. Here we have developed a genetic approach to address the functional relevance of a defined antifungal or antibacterial peptide in the host defense of Drosophila adults. We have expressed AMP genes via the control of the UAS/GAL4 system in imd; spatzle double mutants that do not express any known endogenous AMP gene. Our results clearly show that constitutive expression of a single peptide in some cases is sufficient to rescue imd; spatzle susceptibility to microbial infection, highlighting the important role of AMPs in Drosophila adult host defense. PMID- 11854513 TI - Abnormal B lymphocyte development and autoimmunity in hypoxia-inducible factor 1alpha -deficient chimeric mice. AB - Immune cells are exposed to low oxygen tensions as they develop and migrate between blood and different tissues, but the mechanisms by which lymphocytes adapt to hypoxia are poorly understood. Studies reported here of hypoxia inducible factor 1alpha (HIF-1alpha) in lymphocyte development and functions suggest that it has a critical role in regulation of these processes. HIF-1alpha deficiency in Hif1alpha(-/-) --> Rag2(-/-) chimeric mice results in dramatic and cell lineage-specific defects, which include appearance of abnormal peritoneal B 1-like lymphocytes, with high expression of B220 (CD45) receptor-associated protein tyrosine phosphatase and autoimmunity (accumulation of anti-dsDNA antibodies and rheumatoid factor in serum, deposits of IgG and IgM in kidney and proteinuria) as well as distortions of maturation of B-2 lymphocytes in bone marrow. PMID- 11854514 TI - Pim serine/threonine kinases regulate the stability of Socs-1 protein. AB - Studies of SOCS-1-deficient mice have implicated Socs-1 in the suppression of JAK STAT (Janus tyrosine kinase-signal transducers and activators of transcription) signaling and T cell development. It has been suggested that the levels of Socs-1 protein may be regulated through the proteasome pathway. Here we show that Socs-1 interacts with members of the Pim family of serine/threonine kinases in thymocytes. Coexpression of the Pim kinases with Socs-1 results in phosphorylation and stabilization of the Socs-1 protein. The protein levels of Socs-1 are significantly reduced in the Pim-1(-/-), Pim-2(-/-) mice as compared with wild-type mice. Similar to Socs-1(-/-) mice, thymocytes from Pim-1(-/-), Pim 2(-/-) mice showed prolonged Stat6 phosphorylation upon IL-4 stimulation. These data suggest that the Pim kinases may regulate cytokine-induced JAK-STAT signaling through modulation of Socs-1 protein levels. PMID- 11854515 TI - Synergistic effects of L- and P-selectin in facilitating tumor metastasis can involve non-mucin ligands and implicate leukocytes as enhancers of metastasis. AB - P-selectin facilitates human carcinoma metastasis in immunodeficient mice by mediating early interactions of platelets with bloodborne tumor cells via their cell surface mucins, and this process can be blocked by heparin [Borsig, L., Wong, R., Feramisco, J., Nadeau, D. R., Varki, N. M. & Varki, A. (2001) Proc. Natl. Acad. Sci. USA 98, 3352-3357]. Here we show similar findings with a murine adenocarcinoma in syngeneic immunocompetent mice but involving a different P selectin ligand, possibly a sulfated glycolipid. Thus, metastatic spread can be facilitated by tumor cell selectin ligands other than mucins. Surprisingly, L selectin expressed on endogenous leukocytes also facilitates metastasis in both the syngeneic and xenogeneic (T and B lymphocyte deficient) systems. PL-selectin double deficient mice show that the two selectins work synergistically. Although heparin can block both P- and L-selectin in vitro, the in vivo effect of a single heparin dose given before tumor cells seems to be completely accounted for by blockade of P-selectin function. Thus, L-selectin on neutrophils, monocytes, and/or NK cells has a role in facilitating metastasis, acting beyond the early time points wherein P-selectin mediates interactions of platelet with tumor cells. PMID- 11854516 TI - Marrow stromal cells form guiding strands in the injured spinal cord and promote recovery. AB - Marrow stromal cells (MSC) can be expanded rapidly in vitro and differentiated into multiple mesodermal cell types. In addition, differentiation into neuron like cells expressing markers typical for mature neurons has been reported. To analyze whether such cells, exposed to differentiation media, could develop electrophysiological properties characteristic of neurons, we performed whole cell recordings. Neuron-like MSC, however, lacked voltage-gated ion channels necessary for generation of action potentials. We then delivered MSC into the injured spinal cord to study the fate of transplanted MSC and possible effects on functional outcome in animals rendered paraplegic. MSC given 1 week after injury led to significantly larger numbers of surviving cells than immediate treatment and significant improvements of gait. Histology 5 weeks after spinal cord injury revealed that MSC were tightly associated with longitudinally arranged immature astrocytes and formed bundles bridging the epicenter of the injury. Robust bundles of neurofilament-positive fibers and some 5-hydroxytryptamine-positive fibers were found mainly at the interface between graft and scar tissue. MSC constitute an easily accessible, easily expandable source of cells that may prove useful in the establishment of spinal cord repair protocols. PMID- 11854517 TI - The AF-1 activation-function of ERalpha may be dispensable to mediate the effect of estradiol on endothelial NO production in mice. AB - Two isoforms of estrogen receptor (ER) have been described: ERalpha and ERbeta. The initial gene targeting of ERalpha, consisting in the introduction of a Neo cassette in exon 1 [alphaERKO, hereafter called ERalpha-Neo KO (knockout)], was reported in 1993. More recently, another mouse deficient in ERalpha because of the deletion of exon 2 (ERalphaKO, hereafter called ERalpha-delta2 KO) was generated. In ovariectomized ERalpha-wild-type mice, estradiol (E(2)) increases uterine weight and basal production of endothelial nitric oxide (NO). Both of these effects are abolished in ERalpha-delta2 KO mice. In contrast, we show here that both of these effects of E(2) are partially (uterine weight) or totally (endothelial NO production) preserved in ERalpha-Neo KO. We also confirm the presence of two ERalpha mRNA splice variants in uterus and aorta from ERalpha-Neo KO mice. One of them encodes a chimeric ERalpha protein (ERalpha55), partially deleted in the A/B domain, that was detected in both uterus and aorta by Western blot analysis. The other ERalpha mRNA splice variant codes for an isoform deleted for the A/B domain (ERalpha46), which was detected in uterus of ERalpha-Neo KO, and wild-type mice. This protein isoform was not detected in aorta. The identification of these two N-terminal modified isoforms in uterus, and at least one of them in aorta, probably explains the persistence of the E(2) effects in ERalpha-Neo KO mice. Furthermore, ERalpha-Neo KO mice may help in the elucidation of the specific functions of full-length ERalpha (ERalpha66) and ERalpha46, both shown to be physiologically generated in vivo. PMID- 11854518 TI - Overhauser enhanced magnetic resonance imaging for tumor oximetry: coregistration of tumor anatomy and tissue oxygen concentration. AB - An efficient noninvasive method for in vivo imaging of tumor oxygenation by using a low-field magnetic resonance scanner and a paramagnetic contrast agent is described. The methodology is based on Overhauser enhanced magnetic resonance imaging (OMRI), a functional imaging technique. OMRI experiments were performed on tumor-bearing mice (squamous cell carcinoma) by i.v. administration of the contrast agent Oxo63 (a highly derivatized triarylmethyl radical) at nontoxic doses in the range of 2-7 mmol/kg either as a bolus or as a continuous infusion. Spatially resolved pO(2) (oxygen concentration) images from OMRI experiments of tumor-bearing mice exhibited heterogeneous oxygenation profiles and revealed regions of hypoxia in tumors (<10 mmHg; 1 mmHg = 133 Pa). Oxygenation of tumors was enhanced on carbogen (95% O(2)/5% CO(2)) inhalation. The pO(2) measurements from OMRI were found to be in agreement with those obtained by independent polarographic measurements using a pO(2) Eppendorf electrode. This work illustrates that anatomically coregistered pO(2) maps of tumors can be readily obtained by combining the good anatomical resolution of water proton-based MRI, and the superior pO(2) sensitivity of EPR. OMRI affords the opportunity to perform noninvasive and repeated pO(2) measurements of the same animal with useful spatial (approximately 1 mm) and temporal (2 min) resolution, making this method a powerful imaging modality for small animal research to understand tumor physiology and potentially for human applications. PMID- 11854519 TI - Heterogeneous proliferative potential of occult metastatic cells in bone marrow of patients with solid epithelial tumors. AB - Bone marrow is a major homing site for circulating epithelial tumor cells. The present study was aimed to assess the proliferative capacity of occult metastatic cells in bone marrow of patients with operable solid tumors especially with regard to their clinical outcome. We obtained bone marrow aspirates from 153 patients with carcinomas of the prostate (n = 46), breast (n = 45), colon (n = 33), and kidney (n = 29). Most of the patients (87%) had primary disease with no clinical signs of overt metastases [tumor-node-metastasis (TNM)-stage UICC (Union Internationale Contre le Cancer) I-III]. After bone marrow was cultured for 21 102 days under special cell culture conditions, viable epithelial cells were detected by cytokeratin staining in 124 patients (81%). The cultured epithelial cells harbored Ki-ras2 mutations and numerical chromosomal aberrations. The highest median number of expanded tumor cells was observed in prostate cancer (2,619 per flask). There was a significant positive correlation between the number of expanded tumor cells and the UICC-stage of the patients (P = 0.03) or the presence of overt metastases (P = 0.04). Moreover, a strong expansion of tumor cells was correlated to an increased rate of cancer-related deaths (P = 0.007) and a reduced survival of the patients (P = 0.006). In conclusion, the majority of cancer patients have viable tumor cells in their bone marrow at primary tumor diagnosis, and the proliferative potential of these cells determines the clinical outcome. PMID- 11854520 TI - Molecular specialization of breast vasculature: a breast-homing phage-displayed peptide binds to aminopeptidase P in breast vasculature. AB - In vivo phage display identifies peptides that selectively home to the vasculature of individual organs, tissues, and tumors. Here we report the identification of a cyclic nonapeptide, CPGPEGAGC, which homes to normal breast tissue with a 100-fold selectivity over nontargeted phage. The homing of the phage is inhibited by its cognate synthetic peptide. Phage localization in tissue sections showed that the breast-homing phage binds to the blood vessels in the breast, but not in other tissues. The phage also bound to the vasculature of hyperplastic and malignant lesions in transgenic breast cancer mice. Expression cloning with a phage-displayed cDNA library yielded a phage that specifically bound to the breast-homing peptide. The cDNA insert was homologous to a fragment of aminopeptidase P. The homing peptide bound aminopeptidase P from malignant breast tissue in affinity chromatography. Antibodies against aminopeptidase P inhibited the in vitro binding of the phage-displayed cDNA to the peptide and the in vivo homing of phage carrying the peptide. These results indicate that aminopeptidase P is the receptor for the breast-homing peptide. This peptide may be useful in designing drugs for the prevention and treatment of breast cancer. PMID- 11854521 TI - Erythropoietin prevents motor neuron apoptosis and neurologic disability in experimental spinal cord ischemic injury. AB - The cytokine erythropoietin (EPO) possesses potent neuroprotective activity against a variety of potential brain injuries, including transient ischemia and reperfusion. It is currently unknown whether EPO will also ameliorate spinal cord injury. Immunocytochemistry performed using human spinal cord sections showed abundant EPO receptor immunoreactivity of capillaries, especially in white matter, and motor neurons within the ventral horn. We used a transient global spinal ischemia model in rabbits to test whether exogenous EPO can cross the blood-spinal cord barrier and protect these motor neurons. Spinal cord ischemia was produced in rabbits by occlusion of the abdominal aorta for 20 min, followed by saline or recombinant human (rHu)-EPO (350, 800, or 1,000 units/kg of body weight) administered intravenously immediately after the onset of reperfusion. The functional neurological status of animals was better for rHu-EPO-treated animals 1 h after recovery from anesthesia, and improved dramatically over the next 48 h. In contrast, saline-treated animals exhibited a poorer neurological score at 1 h and did not significantly improve. Histopathological examination of the affected spinal cord revealed widespread motor neuron injury associated with positive terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling in control but not in rHu-EPO-treated animals. These observations suggest both an acute as well as a delayed beneficial action of rHu-EPO in ischemic spinal cord injury. Because rHu-EPO is currently used widely with an excellent safety profile, clinical trials evaluating its potential to prevent motor neuron apoptosis and the neurological deficits that occur as a consequence of ischemic injury are warranted. PMID- 11854522 TI - Iron deficiency and iron excess damage mitochondria and mitochondrial DNA in rats. AB - Approximately two billion people, mainly women and children, are iron deficient. Two studies examined the effects of iron deficiency and supplementation on rats. In study 1, mitochondrial functional parameters and mitochondrial DNA (mtDNA) damage were assayed in iron-deficient (< or =5 microg/day) and iron-normal (800 microg/day) rats and in both groups after daily high-iron supplementation (8,000 microg/day) for 34 days. This dose is equivalent to the daily dose commonly given to iron-deficient humans. Iron-deficient rats had lower liver mitochondrial respiratory control ratios and increased levels of oxidants in polymorphonuclear leukocytes, as assayed by dichlorofluorescein (P < 0.05). Rhodamine 123 fluorescence of polymorphonuclear-leukocytes also increased (P < 0.05). Lowered respiratory control ratios were found in daily high-iron-supplemented rats regardless of the previous iron status (P < 0.05). mtDNA damage was observed in both iron-deficient rats and rats receiving daily high-iron supplementation, compared with iron-normal rats (P < 0.05). Study 2 compared iron-deficient rats given high doses of iron (8,000 microg) either daily or every third day and found that rats given iron supplements every third day had less mtDNA damage on the second and third day after the last dose compared to daily high iron doses. Both inadequate and excessive iron (10 x nutritional need) cause significant mitochondrial malfunction. Although excess iron has been known to cause oxidative damage, the observation of oxidant-induced damage to mitochondria from iron deficiency has been unrecognized previously. Untreated iron deficiency, as well as excessive-iron supplementation, are deleterious and emphasize the importance of maintaining optimal iron intake. PMID- 11854523 TI - Motexafin gadolinium (Gd-Tex) selectively induces apoptosis in HIV-1 infected CD4+ T helper cells. AB - Here, we show that motexafin gadolinium (Gd-Tex), a compound that promotes intracellular oxidative stress, selectively induces apoptosis in HIV-1-infected CD4(+) T cells in IL-2-stimulated cultures of peripheral blood mononuclear cells infected in vitro with HIV-1. This selective induction of apoptosis, which we detect by FACS analysis of intracellular HIV/p24 and concomitant surface and apoptosis marker expression, is abrogated by the glutathione precursor, N-acetyl l-cysteine. Importantly, it occurs at Gd-Tex concentrations that are not cytotoxic to uninfected cells in the culture. These findings suggest that Gd-Tex may have therapeutic utility as an anti-HIV agent capable of selectively targeting and removing HIV-infected cells in an infected host. PMID- 11854524 TI - Genomewide identification of Pseudomonas syringae pv. tomato DC3000 promoters controlled by the HrpL alternative sigma factor. AB - The ability of Pseudomonas syringae pv. tomato DC3000 to parasitize tomato and Arabidopsis thaliana depends on genes activated by the HrpL alternative sigma factor. To support various functional genomic analyses of DC3000, and specifically, to identify genes involved in pathogenesis, we developed a draft sequence of DC3000 and used an iterative process involving computational and gene expression techniques to identify virulence-implicated genes downstream of HrpL responsive promoters. Hypersensitive response and pathogenicity (Hrp) promoters are known to control genes encoding the Hrp (type III protein secretion) machinery and a few type III effector proteins in DC3000. This process involved (i) identification of 9 new virulence-implicated genes in the Hrp regulon by miniTn5gus mutagenesis, (ii) development of a hidden Markov model (HMM) trained with known and transposon-identified Hrp promoter sequences, (iii) HMM identification of promoters upstream of 12 additional virulence-implicated genes, and (iv) microarray and RNA blot analyses of the HrpL-dependent expression of a representative subset of these DC3000 genes. We found that the Hrp regulon encodes candidates for 4 additional type III secretion machinery accessory factors, homologs of the effector proteins HopPsyA, AvrPpiB1 (2 copies), AvrPpiC2, AvrPphD (2 copies), AvrPphE, AvrPphF, and AvrXv3, and genes associated with the production or metabolism of virulence factors unrelated to the Hrp type III secretion system, including syringomycin synthetase (SyrE), N(epsilon) (indole-3-acetyl)-l-lysine synthetase (IaaL), and a subsidiary regulon controlling coronatine production. Additional candidate effector genes, hopPtoA2, hopPtoB2, and an avrRps4 homolog, were preceded by Hrp promoter-like sequences, but these had HMM expectation values of relatively low significance and were not detectably activated by HrpL. PMID- 11854526 TI - The neural basis of intermittent motor control in humans. AB - The basic question of whether the human brain controls continuous movements intermittently is still under debate. Here we show that 6- to 9-Hz pulsatile velocity changes of slow finger movements are directly correlated to oscillatory activity in the motor cortex, which is sustained by cerebellar drive through thalamus and premotor cortex. Our findings suggest that coupling of 6- to 9-Hz oscillatory activity in the cerebello-thalamo-cortical loop represents the neural mechanism for the intermittent control of continuous movements. PMID- 11854525 TI - Murine retroviruses activate B cells via interaction with toll-like receptor 4. AB - Although most retroviruses require activated cells as their targets for infection, it is not known how this is achieved in vivo. A candidate protein for the activation of B cells by either mouse mammary tumor virus (MMTV) or murine leukemia virus is the toll-like receptor 4 (TLR4), a component of the innate immune system. MMTV caused B cell activation in C3H/HeN mice but not in C3H/HeJ or BALB/c (C.C3H Tlr4(lps-d)) congenic mice, both of which have a mutant TLR4 gene. This activation was independent of viral gene expression, because it occurred after treatment of MMTV with ultraviolet light or 2,2'-dithiodipyridine and in azidothymidine-treated mice. Nuclear extracts prepared from the lymphocytes of MMTV-injected C3H/HeN but not C3H/HeJ mice showed increased nuclear factor kappaB activity. Additionally, the MMTV- and Moloney murine leukemia virus envelope proteins coimmunoprecipitated with TLR4 when expressed in 293T cells. The MMTV receptor failed to coimmunoprecipitate with TLR4, suggesting that MMTV/TLR4 interaction is independent of virus attachment and fusion. These results identify retroviral proteins that interact with a mammalian toll receptor and show that direct activation by such viruses may initiate in vivo infection pathways. PMID- 11854527 TI - Heteromultimers of DEG/ENaC subunits form H+-gated channels in mouse sensory neurons. AB - Acidic extracellular solution activates transient H(+)-gated currents in dorsal root ganglion (DRG) neurons. The biophysical properties of three degenerin/epithelial sodium (DEG/ENaC) channel subunits (BNC1, ASIC, and DRASIC), and their expression in DRG, suggest that they might underlie these H(+)-gated currents and function as sensory transducers. However, it is uncertain which of these DEG/ENaC subunits generate the currents, and whether they function as homomultimers or heteromultimers. We found that the biophysical properties of transient H(+)-gated currents from medium to large mouse DRG neurons differed from BNC1, ASIC, or DRASIC expressed individually, but were reproduced by coexpression of the subunits together. To test the contribution of each subunit, we studied DRG from three strains of mice, each bearing a targeted disruption of BNC1, ASIC, or DRASIC. Deletion of any one subunit did not abolish H(+)-gated currents, but altered currents in a manner consistent with heteromultimerization of the two remaining subunits. These data indicate that combinations of two or more DEG/ENaC subunits coassemble as heteromultimers to generate transient H(+) gated currents in mouse DRG neurons. PMID- 11854528 TI - Regulation of Kv1 subunit expression in oligodendrocyte progenitor cells and their role in G1/S phase progression of the cell cycle. AB - Proliferative oligodendrocyte progenitor cells (OPs) express large, delayed outward-rectifying K(+) currents (I(K)), whereas nondividing immature and mature oligodendrocytes display much smaller I(K). Here, we show that up-regulation of I(K) occurs in G(1) phase of the cell cycle in purified cultured OPs and is the result of an RNA synthesis-dependent, selective increase of the K(+) channel subunit proteins Kv1.3 and Kv1.5. In oligodendrocyte cells acutely isolated from developing rat brain, a decrease of cyclin D expression is observed as these cells mature along their lineage. This is accompanied by a decrease in Kv1.3 and Kv1.5 subunit expression, suggesting a role for these subunits in the proliferative potential of OPs in situ. I(K) expressed in OPs in subventricular zone and developing white matter in acutely isolated slice preparations were selectively blocked by antagonists of Kv1.3, illustrating the functional presence of this subunit in situ. Interestingly, Kv1.3 block inhibited S-phase entry of both purified OPs in culture and in tissue slice cultures. Thus, we employ both in vitro and in situ experimental approaches to show that (i) RNA-dependent synthesis of Kv1.3 and Kv1.5 subunit proteins occurs in G(1) phase of the OP cell cycle and is responsible for the observed increase in I(K), and (ii) currents through Kv1.3-containing channels play a crucial role in G(1)/S transition of proliferating OPs. PMID- 11854529 TI - Memory loss in old rats is associated with brain mitochondrial decay and RNA/DNA oxidation: partial reversal by feeding acetyl-L-carnitine and/or R-alpha -lipoic acid. AB - Accumulation of oxidative damage to mitochondria, protein, and nucleic acid in the brain may lead to neuronal and cognitive dysfunction. The effects on cognitive function, brain mitochondrial structure, and biomarkers of oxidative damage were studied after feeding old rats two mitochondrial metabolites, acetyl l-carnitine (ALCAR) [0.5% or 0.2% (wt/vol) in drinking water], and/or R-alpha lipoic acid (LA) [0.2% or 0.1% (wt/wt) in diet]. Spatial memory was assessed by using the Morris water maze; temporal memory was tested by using the peak procedure (a time-discrimination procedure). Dietary supplementation with ALCAR and/or LA improved memory, the combination being the most effective for two different tests of spatial memory (P < 0.05; P < 0.01) and for temporal memory (P < 0.05). Immunohistochemical analysis showed that oxidative damage to nucleic acids (8-hydroxyguanosine and 8-hydroxy-2'-deoxyguanosine) increased with age in the hippocampus, a region important for memory. Oxidative damage to nucleic acids occurred predominantly in RNA. Dietary administration of ALCAR and/or LA significantly reduced the extent of oxidized RNA, the combination being the most effective. Electron microscopic studies in the hippocampus showed that ALCAR and/or LA reversed age-associated mitochondrial structural decay. These results suggest that feeding ALCAR and LA to old rats improves performance on memory tasks by lowering oxidative damage and improving mitochondrial function. PMID- 11854530 TI - Attenuation of thermal nociception and hyperalgesia by VR1 blockers. AB - Vanilloid receptor subunit 1 (VR1) appears to play a critical role in the transduction of noxious chemical and thermal stimuli by sensory nerve endings in peripheral tissues. Thus, VR1 antagonists are useful compounds to unravel the contribution of this receptor to pain perception, as well as to induce analgesia. We have used a combinatorial approach to identify new, nonpeptidic channel blockers of VR1. Screening of a library of trimers of N-alkylglycines resulted in the identification of two molecules referred to as DD161515 [N-[2-(2-(N methylpyrrolidinyl)ethyl]glycyl]-[N-[2,4-dichlorophenethyl]glycyl]-N-(2,4 dichlorophenethyl)glycinamide] and DD191515 [[N-[3-(N,N diethylamino)propyl]glycyl]-[N-[2,4-dichlorophenethyl]glycyl]-N-(2,4 dichlorophenethyl)glycinamide] that selectively block VR1 channel activity with micromolar efficacy, rivaling that characteristic of vanilloid-related inhibitors. These compounds appear to be noncompetitive VR1 antagonists that recognize a receptor site distinct from that of capsaicin. Intraperitoneal administration of both trialkylglycines into mice significantly attenuated thermal nociception as measured in the hot plate test. It is noteworthy that these compounds eliminated pain and neurogenic inflammation evoked by intradermal injection of capsaicin into the animal hindpaw, as well as the thermal hyperalgesia induced by tissue irritation with nitrogen mustard. In contrast, responses to mechanical stimuli were not modified by either compound. Modulation of sensory nerve fibers excitability appears to underlie the peptoid analgesic activity. Collectively, these results indicate that blockade of VR1 activity attenuates chemical and thermal nociception and hyperalgesia, supporting the tenet that this ionotropic receptor contributes to chemical and thermal sensitivity and pain perception in vivo. These trialkylglycine-based, noncompetitive VR1 antagonists may likely be developed into analgesics to treat inflammatory pain. PMID- 11854531 TI - Ca2+ marks: miniature calcium signals in single mitochondria driven by ryanodine receptors. AB - Propagation of cytosolic [Ca(2+)] ([Ca(2+)](c)) signals to the mitochondria is believed to be supported by a local communication between Ca(2+) release channels and adjacent mitochondrial Ca(2+) uptake sites, but the signaling machinery has not been explored at the level of elementary Ca(2+) release events. Here, we demonstrate that [Ca(2+)](c) sparks mediated by ryanodine receptors are competent to elicit miniature mitochondrial matrix [Ca(2+)] signals that we call "Ca(2+) marks." Ca(2+) marks are restricted to single mitochondria and typically last less than 500 ms. The decay of Ca(2+) marks relies on extrusion of Ca(2+) from the mitochondria through the Ca(2+) exchanger, whereas [Ca(2+)](c) sparks decline primarily by diffusion. Mitochondria also appear to have a direct effect on the properties of [Ca(2+)](c) sparks, because inhibition of mitochondrial Ca(2+) uptake results in an increase in the frequency and duration of [Ca(2+)](c) sparks. Thus, a short-lasting opening of a cluster of Ca(2+) release channels can yield activation of mitochondrial Ca(2+) uptake, and the competency of mitochondrial Ca(2+) handling may be an important determinant of cardiac excitability through local feedback control of elementary [Ca(2+)](c) signals. PMID- 11854532 TI - Expression of bitter taste receptors of the T2R family in the gastrointestinal tract and enteroendocrine STC-1 cells. AB - Although a role for the gastric and intestinal mucosa in molecular sensing has been known for decades, the initial molecular recognition events that sense the chemical composition of the luminal contents has remained elusive. Here we identified putative taste receptor gene transcripts in the gastrointestinal tract. Our results, using reverse transcriptase-PCR, demonstrate the presence of transcripts corresponding to multiple members of the T2R family of bitter taste receptors in the antral and fundic gastric mucosa as well as in the lining of the duodenum. In addition, cDNA clones of T2R receptors were detected in a rat gastric endocrine cell cDNA library, suggesting that these receptors are expressed, at least partly, in enteroendocrine cells. Accordingly, expression of multiple T2R receptors also was found in STC-1 cells, an enteroendocrine cell line. The expression of alpha subunits of G proteins implicated in intracellular taste signal transduction, namely Galpha(gust), and Galpha(t)-(2), also was demonstrated in the gastrointestinal mucosa as well as in STC-1 cells, as revealed by reverse transcriptase-PCR and DNA sequencing, immunohistochemistry, and Western blotting. Furthermore, addition of compounds widely used in bitter taste signaling (e.g., denatonium, phenylthiocarbamide, 6-n-propil-2-thiouracil, and cycloheximide) to STC-1 cells promoted a rapid increase in intracellular Ca(2+) concentration. These results demonstrate the expression of bitter taste receptors of the T2R family in the mouse and rat gastrointestinal tract. PMID- 11854533 TI - Analysis of MADS box protein-protein interactions in living plant cells. AB - Over the last decade, the yeast two-hybrid system has become the tool to use for the identification of protein-protein interactions and recently, even complete interactomes were elucidated by this method. Nevertheless, it is an artificial system that is sensitive to errors resulting in the identification of false positive and false-negative interactions. In this study, plant MADS box transcription factor interactions identified by yeast two-hybrid systems where studied in living plant cells by a technique based on fluorescence resonance energy transfer (FRET). Petunia MADS box proteins were fused to either cyan fluorescent protein or yellow fluorescent protein and transiently expressed in protoplasts followed by FRET-spectral imaging microscopy and FRET-fluorescence lifetime imaging microscopy to detect FRET and hence protein-protein interactions. All petunia MADS box heterodimers identified in yeast were confirmed in protoplasts. However, in contrast to the yeast two-hybrid results, homodimerization was demonstrated in plant cells for three petunia MADS box proteins. Heterodimers were identified between the ovule-specific MADS box protein FLORAL BINDING PROTEIN 11 and members of the petunia FLORAL BINDING PROTEIN 2 subfamily, which are also expressed in ovules, suggesting that these dimers play a role in ovule development. Furthermore, the role of dimerization in translocation of MADS box protein dimers to the nucleus is demonstrated, and the nuclear localization signal of MADS box proteins has been mapped to the N terminal region of the MADS domain by means of mutant analyses. PMID- 11854534 TI - Blue light-induced kinetics of H+ and Ca2+ fluxes in etiolated wild-type and phototropin-mutant Arabidopsis seedlings. AB - Ion flux kinetics associated with blue light (BL) treatment of two wild types (WTs) and the phot1, phot2 and phot1/phot2 mutants of Arabidopsis were studied by using the MIFE noninvasive ion-selective microelectrode technique. BL induced significant changes in activity of H(+) and Ca(2+) transporters within the first 10 min of BL onset, peaking between 3 and 5 min. In all WT plants and in phot2 mutants, BL induced immediate Ca(2+) influx. In phot1 and phot1/phot2 mutants, net Ca(2+) flux remained steady. It is suggested that PHOT1 regulates Ca(2+) uptake into the cytoplasm from the apoplast. Changes in ion fluxes were measured from cotyledons of intact seedlings and from the cut top of the hypocotyl of decapitated seedlings. Thus the photoreceptors mediating BL-induced Ca(2+) and H(+) fluxes are present in the rest of the decapitated seedling and probably in the cotyledons as well. The H(+) and Ca(2+) flux responses to BL appear not to be linked because, (i) when changes were observed for both ions, Ca(2+) flux changed almost immediately, whereas H(+) flux lagged by about 1.5 min; (ii) in the Wassilewskija ecotype, changes in H(+) fluxes were small. Finally, wave-like changes in Ca(2+) and H(+) concentrations were observed along the cotyledon-hook axis regardless of its orientation to the light. PMID- 11854535 TI - Cryptic invasion by a non-native genotype of the common reed, Phragmites australis, into North America. AB - Cryptic invasions are a largely unrecognized type of biological invasion that lead to underestimation of the total numbers and impacts of invaders because of the difficulty in detecting them. The distribution and abundance of Phragmites australis in North America has increased dramatically over the past 150 years. This research tests the hypothesis that a non-native strain of Phragmites is responsible for the observed spread. Two noncoding chloroplast DNA regions were sequenced for samples collected worldwide, throughout the range of Phragmites. Modern North American populations were compared with historical ones from herbarium collections. Results indicate that an introduction has occurred, and the introduced type has displaced native types as well as expanded to regions previously not known to have Phragmites. Native types apparently have disappeared from New England and, while still present, may be threatened in other parts of North America. PMID- 11854536 TI - Nuts, nut cracking, and pitted stones at Gesher Benot Ya'aqov, Israel. AB - The Acheulian site of Gesher Benot Ya'aqov (Israel) has revealed a unique association of edible nuts with pitted hammers and anvils. Located in the Dead Sea rift, on the boundary between the Arabian and African plates, the site dates to the Early-Middle Pleistocene, oxygen isotope stage 19. In a series of strata, seven species of nuts, most of which can be cracked open only by a hard hammer, were uncovered. Five of the species are extant terrestrial nuts, and two are aquatic nuts now extinct in the Levant. In addition, the site yielded an assemblage of pitted hammers and anvils similar in pit morphology to those used by chimpanzees and contemporary hunter-gatherers. This is the first time, to our knowledge, that a site has offered both paleobotanical and lithic evidence of plant foods eaten by early hominins and technologies used for processing these foods. The evidence also sheds light on the structure of the community: ethnographic analogies suggest that mixedgender groups may have been active on the shores of paleoLake Hula. PMID- 11854537 TI - The SAP family: a new class of adaptor-like molecules that regulates immune cell functions. AB - EAT-2 is a member of a newly described adaptor-like protein family consisting, at the moment, of EAT-2 and the signaling lymphocytic-activation molecule (SLAM) associated protein (SAP). Both proteins are expressed in immune cells and can bind to members of the SLAM family of immune receptors. However, differences between the two proteins exist. There appears to be little overlap in the types of immune cells that express SAP and EAT-2, and the Src homology 2 (SH2) domain of SAP can bind to unphosphorylated tyrosines, whereas the SH2 domain of EAT-2 cannot. Veillette discusses new findings on the functions of EAT-2 and SAP and how they might regulate signals emanating from the SLAM family of receptors. PMID- 11854541 TI - The weaknesses of science for profit. PMID- 11854538 TI - Infrared-guided laser stimulation of neurons in brain slices. AB - Infrared-guided laser stimulation is a new technique that allows precise and rapid stimulation of visualized neurons in brain slices. Infrared imaging of neurons with a new contrast system is combined with the photolytic release of caged neurotransmitters by an ultraviolet (UV) laser. Addition of caged neurotransmitters to the superfusion medium of neurons in brain slices allows local excitation in the micrometer range with a focused spot of UV light. In this way, the distribution of glutamate or gamma-aminobutyric acid (GABA) receptors on neuronal dendrites can be mapped. Furthermore, this technique can be used to map the connectivity of neuronal networks through the controlled stimulation of neighboring neurons. Because the laser stimulation can be performed much faster than can paired recording, it is also possible to search for synaptic connections between distant neurons that have a low probability of connectivity. PMID- 11854543 TI - Development of the proteasome inhibitor PS-341. AB - Over the last decade, the critical role of the proteasome in cell-cycle regulation has become increasingly apparent. The proteasome, a multicatalytic protease present in all eukaryotic cells, is the primary component of the protein degradation pathway of the cell. By degrading regulatory proteins (or their inhibitors), the proteasome serves as a central conduit for many cellular regulatory signals and, thus, is a novel target for therapeutic drugs. PS-341 is a small molecule that is a potent and selective inhibitor of the proteasome. In vitro and mouse xenograft studies of PS-341 have shown antitumor activity in a variety of tumor types, including myeloma, chronic lymphocytic leukemia, prostate cancer, pancreatic cancer, and colon cancer, among others. Although PS-341 rapidly leaves the vascular compartment, a novel pharmacodynamic assay has shown that inhibition of proteasome-the biologic target-is dose dependent and reversible. These studies provided the rationale for a twice-weekly dosing schedule employed in ongoing clinical studies. Phase I trials in a variety of tumor types have shown PS-341 to be well tolerated, and phase II trials in several hematologic malignancies and solid tumor types are now in progress. Efficacy and safety data from the most advanced of these, a phase II multicenter trial in myeloma, will be available in early 2002. PMID- 11854544 TI - Targeting protein kinase C: new therapeutic opportunities against high-grade malignant gliomas? AB - A large body of evidence suggests that the abnormal phenotype of neoplastic astrocytes, including their excessive proliferation rate and high propensity to invade surrounding tissues, results from mutations in critical genes involved in key cellular events. These genetic alterations can affect cell-surface-associated receptors, elements of signaling pathways, or components of the cell cycle clock, conferring a gain or a loss of relevant metabolic functions of the cells. The understanding of such phenomena may allow the development of more efficacious forms of cancer treatment. Examples are therapies specifically directed against overexpressed epidermal growth factor receptor, hyperactive Ras, excessively stimulated Raf-1, overproduced ornithine decarboxylase, or aberrantly activated cyclin-dependent kinases. The applicability of some of these approaches is now being assessed in patients suffering from primary malignant central nervous system tumors that are not amenable to current therapeutic modalities. Another potentially useful therapeutic strategy against such tumors involves the inhibition of hyperactive or overexpressed protein kinase C (PKC). This strategy is justified by the decrease in cell proliferation and invasion following inhibition of the activity of this enzyme observed in preclinical glioma models. Thus, interference with PKC activity may represent a novel form of experimental cancer treatment that may simultaneously restrain the hyperproliferative state and the invasive capacity of high-grade malignant gliomas without inducing the expected toxicity of classical cytotoxic agents. Of note, the experimental use of PKC-inhibiting agents in patients with refractory high-grade malignant gliomas has indeed led to some clinical responses. The present paper reviews the current status of the biochemistry and molecular biology of PKC, as well as the possibilities for developing novel anti-PKC-based therapies for central nervous system malignancies. PMID- 11854545 TI - Localized adenocarcinoma of the pancreas: the rationale for preoperative chemoradiation. AB - Pancreatic adenocarcinoma is the fifth leading cause of cancer-related death in the U.S. In spite of advancements in surgical treatment, nearly 80% of patients thought to have localized pancreatic cancer die of recurrent or metastatic disease when treated with surgery alone. Therefore, efforts to alter the patterns of recurrence and improve survival for patients with pancreatic cancer currently focus on the delivery of systemic therapy and irradiation before or after surgery. Postoperative adjuvant therapy appears to improve median survival. However, more than one-fourth of patients do not complete planned adjuvant therapy due to surgical complications or a delay in postoperative recovery of performance status. Utilizing a preoperative (neoadjuvant) approach, overall treatment time is reduced, a greater proportion of patients receive all components of therapy, and patients with rapidly progressive disease are spared the side effects of surgery as metastatic disease may be found at restaging following chemoradiation (prior to surgery). This paper examines the factors pertinent to clinical trial design for resectable pancreatic cancer, and carefully reviews the existing data supporting adjuvant and neoadjuvant therapy for potentially resectable disease. PMID- 11854546 TI - Adenoviral gene therapy. AB - As of May 2001, 532 gene therapy protocols had been approved for evaluation in clinical trials; however, only five of those had been evaluated in phase III clinical trials. Among the most commonly used vectors for the delivery of genetic material into human cells are the adenoviruses. Remarkable progress has been made with these vectors in the last decade, but some shortcomings continue to challenge investigators. The newly acquired knowledge of the adenoviral life cycle and the positive outcomes from phase II clinical trials have led to the application of vectors engineered to selectively target tumor tissue under controlled promoters. PMID- 11854547 TI - Breast cancer prevention. AB - Epidemiological, experimental, and clinical data strongly support the possibility that breast cancer will be prevented by using anti-estrogenic interventions in healthy women. Three trials involving over 20,000 women have so far been reported using tamoxifen 20 mg/day or placebo in healthy women to chemoprevent breast cancer. The American National Surgical Adjuvant Breast and Bowel P-1 Project randomized over 13,000 women to take tamoxifen or placebo and showed a 49% reduction in the early incidence of breast cancer. This was associated with a reduction in osteoporotic fractures but increases in the risks of endometrial cancer, cataract, and thromboembolism. The Royal Marsden tamoxifen trial randomized 2,500 women, and the Italian national trial randomized 5,000 women. Interim analyses from these two trials showed no effect on the early incidence of breast cancer. These results, therefore, have not been able to clearly show an overall clinical benefit of giving tamoxifen to healthy women, nor have they shown which women are likely to benefit. Another selective anti-estrogen (SERM), raloxifene, has been used in a clinical trial to prevent osteoporotic fractures in women with low bone mineral density. Annual mammography in this trial has shown an approximate 80% reduction in the early incidence of breast cancer, and further follow-up of this trial continues. New trials in chemoprevention of breast cancer being started or being proposed use luteinizing-hormone-releasing hormone analogues, aromatase inhibitors, and other SERMs. PMID- 11854549 TI - Molecular aspects of the mammalian cell cycle and cancer. AB - Cancer arises mainly from mutations in somatic cells. However, it is not the result of a single mutation, rather, it results from increasing genetic disarray accumulated over time. Tumorigenesis in humans is, therefore, a multistep and age dependent process. The multiple mechanisms and multiple players involved in this process necessitate an understanding of the molecular mechanisms, in order to distinctively classify the tumor sample and to assess the risk and treatment of the disease. PMID- 11854548 TI - The cardiotoxic potential of the 5-HT(3) receptor antagonist antiemetics: is there cause for concern? AB - PURPOSE: To review and evaluate the potential cardiac effects of 5-HT(3) antiemetic treatment in patients who may be predisposed to cardiac complications resulting from malignancy, cytotoxic oncologic regimens, or preexisting comorbid conditions. DESIGN: A literature review was conducted on the negative cardiovascular effects of chemotherapeutic agents, and, more specifically, on the cardiac interactions of the 5-HT(3) receptor antagonists commonly used to treat chemotherapy-induced nausea and vomiting. RESULTS: Clinical studies in healthy subjects have reported electrocardiograph changes following administration of 5 HT(3) receptor antagonists. However, there are limited data on the use of 5-HT(3) antiemetics when administered with cardiotoxic chemotherapy. Nonetheless, the development of significant electrocardiograph changes with some agents may indicate a potential for significant cardiac effects in patients, particularly those who may be predisposed to cardiac complication. CONCLUSIONS: As the predicted human life span increases, clinicians will be treating a larger, older oncology population. Because two of the most common major comorbidities are cardiovascular related, we need to be acutely aware of the toxic effects of chemotherapy, as well as the possible cardiac interaction of supportive agents, specifically the 5-HT(3) antiemetics. Until more data are made available, the best antiemetic option for patients receiving emetogenic and cardiotoxic chemotherapy may be the agent with the fewest apparent cardiac effects. PMID- 11854550 TI - The molecular perspective: restriction endonucleases. PMID- 11854552 TI - Maniacs, malignancy, medicine. PMID- 11854553 TI - Thoughts about September 11, 2001: civilization's resilience and tenacity. PMID- 11854556 TI - September 11, 2001: terrorism external and internal. PMID- 11854557 TI - Endoscopic ultrasonography of the pancreas: new advances. PMID- 11854558 TI - Effect of protein kinase C on glucose-mediated insulin secretion in HIT-T15 cells. AB - OBJECTIVE: To clarify the regulation of protein kinase C on glucose-mediated insulin secretion. MAIN OUTCOME MEASURES: We examined the effect of protein kinase C on the cytosolic free Ca(2+) concentration ([Ca(2+)]i) and the activity of Ca(2+)-activated K(+) channels (K(Ca)-channel) in the insulinoma cell line, HIT-T15. RESULTS: Glucose at a concentration of 10 mmol/L increased the secretion of insulin. This increase was partly inhibited by 1 nmol/L staurosporine, a protein kinase C inhibitor. Staurosporine (1 nmol/L) also attenuated the glucose induced elevations in [Ca(2+)]i. On the contrary, glibenclamide (100 nmol/L) specifically blocked ATP-sensitive K(+) channels, and increased both [Ca(2+)]i and insulin secretion, but staurosporine had no effect on them. Patch clamp studies showed that 10 mmol/L glucose almost completely blocked K(Ca) channel activity, an effect that was reversed by 1 nmol/L staurosporine. Phorbol 12 myristate 13-acetate (1 mmol/L), a protein kinase C activator, also decreased K(Ca) channel activity. CONCLUSIONS: These results indicate that the activation of protein kinase C is involved in the glucose-induced release of insulin by modulating K(+) channel function in HIT-T15 cells. PMID- 11854559 TI - Clinical usefulness of the serum carboxypeptidase B activation peptide in acute pancreatitis. AB - OBJECTIVE: To assess the sensitivity and specificity of the serum carboxypeptidase B activation peptide in diagnosing and determining the severity of acute pancreatitis. PATIENTS: Twenty consecutive patients with acute pancreatitis were studied on admission to the Emergency Room: 11 patients had mild pancreatitis, and 9 patients, severe pancreatitis. Twenty consecutive patients with non-pancreatic acute abdomen and 20 healthy subjects were also studied. MAIN OUTCOME MEASURES: Serum carboxypeptidase B activation peptide was determined using radioimmunoassay. RESULTS: Nineteen of the 20 patients with acute pancreatitis (95.0%) had serum carboxypeptidase B activation peptide concentrations above the upper reference limit, whereas 1 of the 20 patients with non-pancreatic acute abdomen (5.0%) and none of the healthy subjects had serum levels of this protein above the upper reference limit. The serum carboxypeptidase B activation peptide concentrations of patients with severe acute pancreatitis were significantly higher than those of patients with mild acute pancreatitis on the 2(nd) (P=0.044) and 3(rd) days (P=0.028) of the study. The overall sensitivity and specificity of carboxypeptidase B activation peptide in assessing the severity of acute pancreatitis were 84.6% and 59.4%, respectively. CONCLUSIONS: Serum carboxypeptidase B activation peptide may be used simultaneously both to diagnosis and assess the severity of acute pancreatitis on admission to the Emergency Room. PMID- 11854560 TI - Chronic pancreatitis: relationship to acute pancreatitis and pancreatic cancer. PMID- 11854561 TI - Pancreatic head mass: what can be done? Classification: the pathological point of view. PMID- 11854562 TI - Pancreatic head mass: what can be done? Classification: the clinical point of view. AB - Surgeons frequently find pancreatic head mass when operating. The obvious difficulty is to make the correct preoperative differential diagnosis between chronic pancreatitis and pancreatic tumor. The first step is to reach a diagnosis, with some certainty, prior to the operation. The second step in the case of a tumor is the accurate staging and deciding whether or not it is resectable. On the one hand, time and cost must be considered; on the other hand, the therapy must be decided. Obtaining information about the characteristics of the pancreatic disease (nature, size, exact location) and establishing the tissue diagnosis preoperatively may simplify the decision to operate and the operation itself. In the case of chronic pancreatitis, the aim of the operation is to eliminate pain and other symptoms, while in the case of cancer, the purpose is to remove the malignant tissue. In most patients, it is possible to identify the disease on the basis of previous examinations together with preoperative diagnostic techniques such as exploration, palpation and fine-needle aspiration biopsy. Chronic pancreatic head mass should be operated upon with Beger s or Frey s procedure while pancreatic head tumors should be treated by means of head resection with the aim of preserving the pylorus or the Whipple procedure may be used. When the diagnosis is in doubt, a radical approach is thought to be best. Our conclusion is that there is no diagnostic method capable of making a definitive differential diagnosis as to the nature of the pancreatic head mass. Further study is required as to the extent to which differential diagnosis should be investigated. PMID- 11854563 TI - Pancreatic head mass: what can be done? Diagnosis: ultrasonography. PMID- 11854564 TI - Pancreatic head mass: what can be done? Diagnosis: computed tomography scan. AB - The diagnosis of different pancreatic diseases has recently become a recurrent problem. In cases with pancreatic head mass the main question is the differentiation between malignant and benign lesions. When a neoplasm is suspected, the main task is to judge operability. The usefulness of computed tomography imaging in the evaluation of pancreatic carcinoma has been well established. In this article the authors discuss the possibilities of computed tomography (CT) in diagnostic work-up. PMID- 11854565 TI - Pancreatic head mass: what can be done? Diagnosis: magnetic resonance imaging. PMID- 11854566 TI - Pancreatic head mass: what can be done? Diagnosis: ERCP and EUS. PMID- 11854567 TI - Pancreatic head mass: what can be done? Diagnosis: angiography. PMID- 11854568 TI - Pancreatic head mass, what can be done? Diagnosis: laboratory. PMID- 11854569 TI - Pancreatic head mass, what can be done? Diagnosis: cytology. PMID- 11854570 TI - Pancreatic head mass: what can be done? Diagnosis: laparoscopy. PMID- 11854571 TI - Pancreatic head mass: what can be done? Diagnosis: surgery. PMID- 11854572 TI - Pancreatic head mass: how can we treat it? Acute pancreatitis: conservative treatment. PMID- 11854573 TI - Pancreatic head mass: how can we treat it? Acute pancreatitis: surgical treatment. PMID- 11854574 TI - Pancreatic head mass: how can we treat it? Chronic pancreatitis: conservative treatment. PMID- 11854575 TI - Pancreatic head mass, how can we treat it? Chronic pancreatitis: surgical treatment. PMID- 11854576 TI - Pancreatic head mass: how can we treat it? Tumor: conservative treatment. PMID- 11854577 TI - Pancreatic head mass: how can we treat it? Tumor: surgical treatment. AB - Pancreatic carcinoma is a devastating disease. Untreated 5-year survival is 0%. The only possibility of being cured is given by surgical removal of the tumor. Pancreatoduodenectomy previously involved high morbidity and mortality rates until it was postulated that palliation gave better results. Today, morbidity and mortality rates have been decreased to an acceptable level, mortality rates in specialized centers being under 5%. Prognostic factors determining survival were found to be the size of the tumor, grade, lymph node involvement and stage. In order to be able to compare results of the different centers, standardization of the surgical technique is mandatory. It is unanimously accepted that in order to improve survival in pancreatic carcinoma, the radicality of the surgical procedure should be increased to include lymphadenectomy. Postoperative adjuvant therapy could also be a determinant factor. Prospective randomized clinical trials will give an answer to these still unanswered questions. PMID- 11854578 TI - Science without logic. PMID- 11854581 TI - The management of muscle strain injuries: an early return versus the risk of recurrence. PMID- 11854582 TI - Summary and agreement statement of the 1st International Symposium on Concussion in Sport, Vienna 2001. PMID- 11854583 TI - Effect of mild brain injury on an instrumented agility task. AB - OBJECTIVE: To identify an objective evaluation technique to guide return-to-play decisions following mild brain injury (MBI) using an agility task that incorporates cognitive and motor performance. DESIGN: Prospective, controlled, repeated measures study. SETTING: Collegiate athletic training facility. PARTICIPANTS: 84 athletes (25 male rugby players, 34 female rugby players, 25 male ice hockey players) from 3 intercollegiate club teams participated. 9 athletes who suffered MBI during their competitive seasons and 9 matched controls completed the entire study protocol. MAIN OUTCOME MEASURES: Time to complete an agility task on the Cybex Reactor and a postconcussive symptoms scale score were assessed in all subjects during the preseason. Injured subjects and matched controls were also assessed 1, 3, 5, and 10 days postinjury. RESULTS: Repeated measures analyses of variance revealed significant differences between injured subjects and controls in postconcussion symptoms, but not agility performance. Post hoc testing revealed that injured subjects reported significantly more postconcussion symptoms on Day 1 after injury. CONCLUSION: The methods of agility assessment used in this study appear to not be sensitive enough to detect functional impairment following MBI. PMID- 11854584 TI - Baseline injury risk factors for runners starting a marathon training program. AB - OBJECTIVE: To describe the baseline characteristics of participants starting a 25 week marathon training program, and their relationship to injury risk factors. DESIGN: Prospective survey. SETTING: Community-based marathon training program. PARTICIPANTS: 1,548 of 2,314 registrants for the Houston Fit Marathon Training Program (mean age 35.8 +/- 9.3 years, mean body mass index 24.3 +/- 3.9, 63% female). INTERVENTION: 4-page survey administered at registration. MAIN OUTCOME MEASURE: Running experience, training practices, demographics, chronic medical problems and previous injuries. RESULTS: Females were more likely to be classified as underweight and males as overweight or obese (p < 0.0001). The mean number of years of running experience was 6.2 +/- 6.2. Most (87.5%) planned to train at a 9-minute mile or slower pace. 52.3% of the participants had not previously trained for a marathon. In the 3 months prior to starting the program 16.1% had been sedentary. Those runners who had not previously completed a marathon and not previously trained with Houston Fit had a higher prevalence of baseline training techniques that could be risk factors for injury. During the previous 3 years, 38.1% reported having an injury, and 35% of all injuries were still symptomatic at the start of the program. CONCLUSIONS: Training programs for the marathon attract more female athletes and those with less running and marathon experience. The prevalence of being overweight or obese is 35.6%. 16.1% are sedentary during the 3 months before starting this program. Training programs must take measures to establish baseline fitness, to educate on injury prevention training techniques, and to set appropriate fitness goals. CLINICAL RELEVANCE: The research study shows that many patients wanting to start a marathon training program are relatively untrained and inexperienced with reference to endurance running. These patients will need special care and education so as to minimize injury and maximize the effect on their physical activity habits. PMID- 11854585 TI - Eye injuries in women's lacrosse players. AB - OBJECTIVE: To determine the prevalence of eye injuries among intercollegiate women's lacrosse players as well as the level of play, specific play settings, and particular mechanisms associated with these injuries. DESIGN: Anonymous, retrospective, survey. SETTING: Intercollegiate lacrosse tournament. PARTICIPANTS: Collegiate and postcollegiate women's lacrosse players. MAIN OUTCOME MEASURES: Self-reports of eye injuries, injury circumstances, treatment received, playing time missed, and continuing problems were recorded. RESULTS: Surveys were collected from 667 players from 34 teams (83% of players listed on rosters). A total of 125 injuries were reported among 84 players (12.6%). Injuries occurred equally in high school and college. Athletes were injured most frequently in games (41 injuries) and practices (38 injuries), with 17 players injured during the off-season. Several mechanisms of injury were noted. Fifty three women sustained injuries requiring medical attention. Of those athletes injured, 23 missed 1-5 days and 9 players missed more than 5 days. Four athletes (4.8%) reported residual problems from an eye injury sustained while playing lacrosse. CONCLUSIONS: Eye injuries are occurring more frequently than current surveillance data suggest. Because injuries are seen at all levels of play, in many different situations, and by multiple mechanisms, mandatory eyewear is needed to prevent serious eye injuries in women's lacrosse. PMID- 11854586 TI - Effect of experience on rodeo injury. AB - OBJECTIVES: To document injury rates, severity, and relative risk during five competitive seasons of Canadian professional rodeo, between experienced (saddle bronc [SB], bareback [BB], and bull riders [BR]) and inexperienced (novice saddle bronc [NSB], novice bareback [NBB], and boys' steer riders [BSR]) rough stock competitors. DESIGN: Prospective cohort study. SETTING: Canadian professional rodeo competition. SUBJECTS: Experienced competitors included professional cowboys from Australia, Brazil, New Zealand, the United States, and Canada. Inexperienced competitors included cowboys from Canada and the United States. METHODS: Data was gathered prospectively at 63 of 323 professional rodeos in Canada from 1995 to 1999, constituting 30.8% of all professional rodeo performances during this time period. Injury data (severity and body part affected) was included when the injury occurred to a registered contestant, at a Canadian professional rodeo, at which the Canadian Professional Rodeo Sport Medicine Team was officially present and providing services. Data were collected by certified athletic therapists. MAIN RESULTS: Inexperienced rough stock competitors had a lower overall rate of injury in comparison to experienced competitors. In addition, inexperienced rough stock competitors had a lower injury rate of severe injuries, and a lower rate of injury to most body parts when compared with experienced competitors. Inexperienced competitors had a high rate of injury to the hand, wrist, and forearm. Most of these injuries to inexperienced contestants occurred to NBB (46%) and BSRs (31%). The relative risk of injury to inexperienced competitors did not differ from experienced competitors in the horse riding events (NSB vs. SB, NBB vs. BB), but the relative risk of injury to BSRs was one-half that of BR (0.49). CONCLUSIONS: Inexperienced competitors in rodeo rough stock events do not have increased rates of severe injury, or of injury to specific body parts (in general). Inexperienced competitors do have a greater rate of injury to the arm, hand, and wrist. The relative risk of BSR is one-half the risk of BR. PMID- 11854587 TI - Practical management of patellofemoral pain. PMID- 11854588 TI - Injuries to the peroneal nerve in professional hockey. PMID- 11854589 TI - Subacute bacterial endocarditis in a footballer--not a sports injury. PMID- 11854590 TI - Achilles tendinopathy during pregnancy. PMID- 11854591 TI - Position Statement. Violence and injury in ice hockey. AB - Ice hockey is a sport enjoyed by many men and women at the spectator and participant level. It is played with high intensity and often involves body contact. Although the women's games is far from injury free, it is the men's game that has drawn criticism for excessive violence. Much attention has been drawn to the serious injuries that have occurred in ice hockey, specifically spinal injuries, concussions, and eye injuries. Many such injuries are the result of illegal and violent acts such as checking from behind or a deliberate high stick. Because of this, some medical organizations have called for changes in the sport, such as minimum age requirements for body-checking. As a practical matter such changes are unlikely to be accepted by hockey governing boards. Many of those involved in the sport consider body-checking a fundamental component of the game. Furthermore, a distinction needs to be made between any kind of injury and a serious, catastrophic injury. For example, although a recent study found that body-checking accounted for up to 38% of ice hockey injuries, none were of the catastrophic type. With respect to catastrophic injuries such as spinal cord trauma or a blinded eye, legal body-checking accounts for significantly less than illegal body-checking (e.g., checking from behind) or violent stick work. To reduce serious injury in ice hockey, we offer 10 recommendations, key among them automatic game suspensions for certain rules violations, and recognition of the coach as the most important figure in promoting a clean, safe game. PMID- 11854592 TI - The role of impact forces and foot pronation: a new paradigm. PMID- 11854595 TI - Immunoglobulin gene rearrangement analysis in composite hodgkin disease and large B-cell lymphoma: evidence for receptor revision of immunoglobulin heavy chain variable region genes in Hodgkin-Reed-Sternberg cells? AB - Immunoglobulin heavy chain gene (IgH) rearrangement was studied in a patient showing the occurrence of classical Hodgkin disease and large B-cell lymphoma (LBCL) in the same lymph node. The VHDHJH region was amplified by polymerase chain reaction, the template being the DNA extracted from single Hodgkin and Reed Sternberg and LBCL cells, microdissected on hematoxylin-eosin-stained sections by laser capture. A repeated VH4DH3JH4 segment was found in Reed-Sternberg cells, whereas a repeated VH3DH3JH4 segment was observed in LBCL cells. Rearranged VH genes carried somatic mutations in both populations, indicating a common germinal center cell origin. The IgH rearrangement found in clonally related Reed Sternberg cells differed from the one of LBCL cells in the VH region but showed the same JH and DH segments with no variation from the respective germline sequence. The DH-JH junction is the first immunoglobulin gene segment rearranged in precursor B cells. Because the possibility of secondary Ig gene rearrangement in peripheral lymphoid organs has recently been reported, in the patient described here Reed-Sternberg and LBCL cells might originate from a common precursor in which secondary VH replacement took place during the germinal center reaction, giving rise to two different clonally related lymphomas. PMID- 11854597 TI - Molecular detection of the synovial sarcoma translocation t(X;18) by real-time polymerase chain reaction in paraffin-embedded material. AB - The t(X;18) translocation is known to be a useful marker for the diagnosis of synovial sarcoma. In this study, the authors describe a new real-time reverse transcriptase-polymerase chain reaction (RT-PCR) method to detect SYT/SSX fusion transcripts using paraffin-embedded and frozen tumor specimens. A series of 38 soft tissue sarcomas were analyzed. Diagnosis was based on clinical, histologic, and immunohistochemical examination. The fusion transcripts were detected in 16 of 17 synovial sarcoma samples (the 17th sample was not suitable for molecular analysis). No t(X;18)-fusion transcript was PCR-amplified in the 21 nonsynovial sarcoma mesenchymal tumors. Therefore, real-time PCR amplification appears to be a powerful, rapid, specific, and sensitive technique that can be used routinely to diagnose the synovial sarcoma t(X;18) translocation. In addition, the t(X;18) can be detected not only on frozen but also on paraffin-embedded tumor samples. PMID- 11854596 TI - Atypical pleomorphic extraosseous ewing tumor/peripheral primitive neuroectodermal tumor with unusual phenotypic/genotypic profile. AB - A pleomorphic undifferentiated tumor primarily located in the retroperitoneum with a phenotype compatible with an extraosseous Ewing tumor/peripheral primitive neuroectodermal tumor (ET/pPNET) pattern and unusual molecular features is described. Immunohistochemically, HBA-71 (CD99/mic2) and several neural markers were intensively expressed together with scattered cells expressing carcinoembryonic antigen (CEA). Short-term culture showed biphasic neuroblastic and epithelioid cell populations, with the latter expressing germ cell markers (CEA, alpha-fetoprotein, and the beta-subunit of chorionic gonadotrophin). Conventional cytogenetics displayed several chromosomic rearrangements, especially a complex translocation t(17,2,22,13) (q21::q11-->q33::q12- >q13::q14). These structural abnormalities were confirmed using fluorescence in situ hybridization analysis. Molecular studies revealed EWS-FEV fusion transcripts (exon 7 of the EWS gene and exon 2 of the FEV gene). In addition, a new p53 mutation not previously reported in ET/pPNET involving exon 5 codon 138: GCC to GAC (Ala/Asp) was detected. In our case, we emphasize the presence of atypical features not only from the phenotypic point of view but also at the genetic level as well as the value of detecting such markers in the differential diagnosis with other abdominal pleomorphic tumors. PMID- 11854598 TI - PTEN mutation is rare in chondrosarcoma. AB - Chondrosarcoma is the second most common primary malignant neoplasm of bone in adults, but the major genetic events involved in the progression of this often fatal cancer remain to be elucidated. Loss of heterozygosity of chromosome 10q has been reported in 67% of chondrosarcoma. The tumor suppressor gene PTEN is located on chromosome 10q, specifically 10q23, raising the possibility that the loss of PTEN function is responsible for some chondrosarcomas. The authors examined 40 chondrosarcoma tumors and tumor-derived cell lines for alterations in PTEN. Only one mutation resulting in a truncated PTEN protein was detected, which was in a metastasized extraskeletal myxoid chondrosarcoma. Thus, mutated PTEN is an uncommon event in the development of chondrosarcoma. The high frequency of loss of heterozygosity on 10q suggests the presence of additional tumor suppressor genes at these loci. PMID- 11854599 TI - Lack of association between hepatitis C viral RNA in serum and liver and histologic gradings: a study on Irish anti-D-treated patients. AB - In this study the authors applied a reverse transcription-polymerase chain reaction (RT-PCR) assay to detect hepatitis C virus (HCV) RNA in 15 frozen liver biopsy samples from anti-D-treated patients. They also correlated the presence or absence of HCV RNA in the serum and liver of each patient with their histologic gradings. RNA was extracted from 36 frozen liver biopsy samples. These included 15 liver biopsy samples from patients infected with HCV through contamination of anti-D blood products. Three of these 15 anti-D-treated patients were receiving alpha-interferon treatment at the time of liver biopsy. Nine frozen liver biopsy samples from patients with a history of intravenous drug abuse were included as positive controls. HCV-negative frozen liver biopsy samples from 12 noninfected patients were used as negative controls. RNA was also extracted from six frozen skin biopsy specimens to check for cross-contamination of samples. Eleven of 15 anti-D-treated patients were HCV RNA positive by RT-PCR, with 100% correlation between HCV RNA in the serum and liver. The nine frozen liver biopsy samples from the intravenous drug abuse patients (positive controls) were also RT-PCR positive for HCV RNA. The 12 noninfected samples and the negative control biopsy samples were negative for HCV. Twenty-seven percent of the recombinant immunoblot assay positive patients were serum and liver HCV RNA negative. HCV-positive patients receiving alpha-interferon therapy at the time of biopsy had cleared the virus from the serum and the liver. There was no correlation between the presence or absence of serum and liver HCV RNA with the histologic grading. This lack of correlation shows clearly the importance of histopathologic evaluation of liver biopsy samples in monitoring HCV-associated liver disease progression. In addition, this finding indicates that one cannot rely only on the presence or absence of HCV RNA in either serum or liver tissue as a parameter in monitoring HCV-associated liver disease progression in this unique cohort of anti-D-treated patients. PMID- 11854600 TI - Expression of survivin messenger RNA correlates with poor prognosis in patients with hepatocellular carcinoma. AB - Suppression of apoptosis is important for carcinogenesis and tumor growth. Recent studies revealed that survivin not only inhibited apoptosis but also accelerated cancer cell proliferative activity. To investigate the prognostic role of expression of the antiapoptosis gene, survivin, in hepatocellular carcinoma (HCC), the authors analyzed the correlation between the expression pattern of survivin messenger RNA (mRNA) and clinicopathologic findings of patients. Tissues were obtained by surgical resection of livers from 51 patients with HCC and 6 patients without HCC. Expression of survivin mRNA was evaluated using reverse transcription-polymerase chain reaction in 51 tumors, 51 adjacent histologically noncancerous livers, and 6 normal livers. Survivin protein expression was evaluated using Western blotting, and apoptotic cancer cells were detected by immunostaining with polyclonal rabbit anti-single-stranded DNA. Survivin mRNA expression was detected in 21 of 51 (41%) tumors, 2 of 51 (4%) noncancerous livers, and none of the 6 normal livers. Survivin mRNA expression did not correlate with tumor size or stage of HCC. Percentage of apoptotic cancer cells of 30 survivin mRNA-negative tumors (5.2 +/- 3.4%) was significantly higher than that of 21 survivin mRNA-positive tumors (2.2 +/- 2.3%, P = 0.0019). The disease free 5-year survival rate of 21 patients positive for survivin mRNA (19%) was significantly poorer than that of 30 patients negative for survivin mRNA (39%, P = 0.0148). Survivin mRNA was detected in 57% (17/30) patients with HCC recurrence but in only 19% (4/21) of patients without recurrence (P = 0.0072). These results indicated that survivin mRNA expression could be used as an independent prognostic factor for patients with HCC after hepatectomy. PMID- 11854601 TI - Molecular stability of DNA typing short tandem repeats in the mammary tree of patients with breast cancer. AB - Archival pathologic specimens are a rich source for the studies of hereditary diseases, cancer genetics, and identification cases in forensic science. In this study, the intraindividual consistency of eight identifying microsatellite polymorphisms (i.e., HMTH01, vWFA31, F13A, MITMH26, FES-FPS, CD4, TPOX, CSF1PO)in a cohort of 40 patients with invasive breast carcinoma were analyzed. Nests of cancer and adjacent morphologically normal ductal-lobular structures (TDLUs) were microdissected as discrete regions from hematoxylin-eosin-stained slides. As controls for each case, DNA templates were prepared from TDLUs located in nontumor quadrants and from unaffected breast skin. Over 1,400 carefully controlled PCR reactions were reviewed, and no evidence was found for microsatellite mismatches among intraindividual cancer and control DNAs. The negative results, supported by validation experiments, strongly argue that alterations of simple repeats are rare somatic events during the onset and progression of breast cancer. This study suggests that PCR artifacts may be a relevant cause of misdiagnosis of microsatellite instability in human sporadic cancer. PMID- 11854602 TI - Use of interphase fluorescence in situ hybridization as a powerful diagnostic tool in cytology. AB - Interphase fluorescence in situ hybridization (I-FISH) using labeled nucleic acid probes detects chromosomal and genetic aberrations at a cellular level. I-FISH is a relatively fast and sensitive technique for evaluating a large number of cells and revealing more specific information than other techniques. It has been proven to be an invaluable molecular test in cytologic analyses for the detection of subtle genetic alterations that correlate with disease progression. In this postgenomic era, with the draft of the human genome available and expansion of the knowledge of tumor-specific genetic changes, the application of I-FISH probes in cytologic analysis should be of great value in the early detection, risk assessment, and monitoring of therapy efficacy in cancer. Here, we outline the principle of the I-FISH procedure, present suggestions to efficiently analyze cytologic materials, provide examples of practical applications, and discuss new aspects of the technique. PMID- 11854603 TI - Lactobacillus plantarum 299V in the treatment and prevention of spontaneous colitis in interleukin-10-deficient mice. AB - Interleukin (IL)-10-deficient (IL-10-/-) mice develop colitis under specific pathogen-free (SPF) conditions and remain disease free if kept sterile (germ free [GF]). We used four different protocols that varied the time-points of oral administration of Lactobacillus plantarum 299v (L. plantarum) relative to colonization with SPF bacteria to determine whether L. plantarum could prevent and treat colitis induced by SPF bacteria in IL-10-/- mice and evaluated the effect of this probiotic organism on mucosal immune activation. Assessment of colitis included blinded histologic scores, measurements of secreted colonic immunoglobulin isotypes, IL-12 (p40 subunit), and interferon (IFN)-gamma production by anti-CD3-stimulated mesenteric lymph node cells. Treating SPF IL-10 /- mice with L. plantarum attenuated previously established colonic inflammation as manifested by decreased mucosal IL-12, IFN-gamma, and immunoglobulin G2a levels. Colonizing GF animals with L. plantarum and SPF flora simultaneously had no protective effects. Gnotobiotic IL-10-/- mice monoassociated with L. plantarum exhibited mild immune system activation but no colitis. Pretreatment of GF mice by colonization with L. plantarum, then exposure to SPF flora and continued probiotic therapy significantly decreased histologic colitis scores. These results demonstrate that L. plantarum can attenuate immune-mediated colitis and suggest a potential therapeutic role for this agent in clinical inflammatory bowel diseases. PMID- 11854604 TI - Infliximab treatment for Crohn's disease: one-year experience in a Dutch academic hospital. AB - The aim of this study was to report the 1-year clinical experience with infliximab treatment for Crohn's disease (CD) in the Netherlands. All 73 CD patients receiving infliximab infusions were prospectively followed during 1 year after the drugs' registration in the Netherlands. Clinical response and adverse events were assessed for both active luminal disease as well as fistulous disease. A total of 212 infusions were administered to 57 patients with active luminal CD and 16 patients with fistulous CD. The mean duration between infusions was 60 days. In 17% of patients, adverse events were recorded, of which one was serious. The response rate was 81% in active luminal CD and 87% in fistulous disease. Response rates were highest in patients receiving concomitant methotrexate as maintenance therapy. Steroids could successfully be tapered off in 73% of responding luminal CD patients and 100% of responding CD patients with fistulae. Eleven patients showed a loss of response to continuous infliximab readministration. Our clinical experience with infliximab for active luminal and fistulous CD showed that the administration is safe, effective, and has high steroid-sparing efficacy. Higher response rates were seen with methotrexate as concomitant medication. PMID- 11854606 TI - Toxicity of mycophenolate mofetil in patients with inflammatory bowel disease. AB - Trials of mycophenolate mofetil (MMF) in inflammatory bowel disease (IBD) suggest that it may be useful in patients intolerant of azathioprine. We examined the safety and efficacy of MMF in IBD patients intolerant of or unresponsive to azathioprine. Twelve patients [seven with Crohn's disease (CD); seven women; mean age 40 years, range 14-76 years] were treated with MMF 500 mg b.i.d. for a mean of 12.5 weeks. Intolerance was defined as the development of side effects that resolved on discontinuing MMF. Improvement was described as symptomatic improvement, decreased steroid use, or disease entering endoscopic remission. Four patients responded with symptomatic improvement and reduced steroids or mesalazine requirement. Three patients developed headache, nausea, or arthralgia. Three patients developed profuse bloody diarrhea, and in two cases with previously quiescent ulcerative colitis (UC), the source was shown to be ulcers in a drug-induced colitis with histologic features similar to those previously reported in four renal transplant patients on MMF. There is no clear evidence of efficacy of MMF in the treatment of IBD, and its use in this condition should be confined to a randomized controlled trial. Moreover, as patients with UC may be unduly prone to colonic injury, MMF may not be a suitable drug for its treatment. PMID- 11854605 TI - Risk factors for low bone density in Crohn's disease. AB - Osteopenia and osteoporosis are prevalent in patients with Crohn's disease (CD). We conducted a cross-sectional study on consecutive patients with CD to assess the prevalence and factors associated with low bone mass density (BMD). One hundred sixty-eight patients with CD were evaluated. Baseline demographics, medical and surgical history, calcium intake, physical activity, steroid use, Harvey Bradshaw Index, blood and urine tests, and dual-energy X-ray absorptiometry were obtained. Sixty-seven (40%) and seventy-five (45%) patients had osteopenia of the femur and spine, respectively. Ten to 11% of patients had osteoporosis. Of the 40 patients who never used steroids, 19 (48%) had osteopenia of the femur and 12 (30%) of the spine. Significant associations were found between BMD and age, body mass index, and serum magnesium. Lifetime steroid use was a weaker predictor of bone loss. Duration of disease did not correlate with BMD when adjusted for age. At follow-up at a mean of 2 years, BMD declined in the femur but not the spine. However, those with ongoing steroid use had lower spine BMD. A significant number of patients with CD have osteopenia. Age was the most important predictor of bone loss. Significant proportion of steroid naive patients had osteopenia, which implies that mechanisms other than steroid use are also involved in bone loss in CD. Disease activity, systemic inflammation, and hormonal and genetic factors may all be important determinants of bone loss in CD. PMID- 11854607 TI - Autoimmune hemolytic anemia associated with Crohn's disease. AB - Autoimmune hemolytic anemia (AHA) is a well-recognized entity associated with ulcerative colitis. Only a single case of AHA previously has been reported in association with Crohn's disease. We describe the first case of this association in a child with perinuclear antineutrophil cytoplasmic antibody-positive Crohn's disease. PMID- 11854609 TI - Management of internal fistulas in Crohn's disease. AB - Internal fistulas occur in 5-10% of patients with Crohn's disease. The clinical presentation of each of the three main types of internal fistulas--enteroenteric, enterovaginal, and enterovesical fistulas--is important in determining the best management. Asymptomatic fistulas usually require no treatment, but fistulas that cause severe or persistent symptoms necessitate intervention. Previously regarded as a surgical condition requiring resection, some internal fistulas are amenable to a more conservative approach involving medical therapy, surgical repair, or both. So far, there have not been any prospective studies designed specifically to assess the efficacy of a medical treatment of internal fistulas, and information about treatment results is gleaned from trials in which patients with internal fistulas have been included and from retrospective reports. Drugs that have been reported to close internal fistulas partially or completely include azathioprine, 6-mercaptopurine, mycophenolate mofetil, cyclosporine A, tacrolimus, and infliximab. Reparative surgical techniques include transrectal and transvaginal mucosal advancement flaps, cutaneous advancement flap, and anal stricturectomy in combination with a rectal mucosal advancement sleeve. Prospective trials of medical therapy and combination medical and surgical therapy for internal fistulas are needed to provide evidence to support the use of these new therapeutic approaches. PMID- 11854608 TI - Metastatic Crohn's disease of the forehead. AB - BACKGROUND: Metastatic Crohn's disease (CD) involves the presence of cutaneous granuloma distant from the intestinal lesions related to the disease, usually observed in colonic CD. CASE HISTORY: A 35-year-old female with a permanent ileostomy following proctocolectomy for CD presented in 1999 with a 2-month history of an unusual skin lesion of the forehead. A diagnosis of CD of the ileum, colon, and rectum had been made in 1994. In 1997, a proctocolectomy with ileostomy was performed due to fistulizing severe refractory disease. Microscopic aspects of the intestinal lesions showed deep and fissuring ulcers. After surgery, she went into remission, and a small bowel follow-up in 1999 showed no recurrence, when she presented with the skin lesion of the forehead. MICROSCOPIC DATA: Histological analysis of endoscopical and surgical intestinal specimens showed chronic granulomatous inflammation of the ileum, colon, and rectum, confirming the diagnosis of CD. The forehead skin biopsy was examined by three independent histopathologists. The lesion was composed of numerous small granulomas (Ziehl-Nielsen negative), with no foreign bodies, mainly composed of CD68-positive and periodic acid Schiff-negative monocytes. Despite the low number of lymphocytes, the macroscopical and microscopical aspect of the forehead lesion, together with the clinical history, led to a diagnosis of rare metastatic CD of the forehead. CONCLUSIONS: This case report describes the development of an unusual granulomatous skin lesion of the forehead in a patient with established CD showing no postoperative recurrence. PMID- 11854610 TI - Etiopathogenesis of inflammatory bowel disease: the importance of the pediatric perspective. AB - Inflammatory bowel disease (IBD) is now recognized as a common chronic disease affecting children and adolescents. This article will review recent advances made in the fields of genetics, epidemiology, gut ecology, and immunology regarding the etiopathogenesis of IBD, with particular emphasis on the contributions made by pediatric studies. Areas where further study of the pediatric age group would be beneficial will be highlighted. PMID- 11854611 TI - Medical management of fulminant colitis. PMID- 11854612 TI - Fulminant colitis: the case for operative treatment. PMID- 11854614 TI - Application of genome-wide gene expression profiling by high-density DNA arrays to the treatment and study of inflammatory bowel disease. AB - Identification of factors involved in the initiation, amplification, and perpetuation of the chronic immune response and the identification of markers for the characterization of patient subgroups remain critical objectives for ongoing research in inflammatory bowel disease (IBD). The Human Genome Project and the development of the expressed sequence tag (EST) clone collection and database have made possible a new revolution in gene expression analysis. Instead of measuring one or a few genes, parallel DNA microarrays are capable of simultaneously measuring expression of thousands of genes, providing a glimpse into the logic and functional grouping of gene programs encoded by our genome. Applied to clinical specimens from affected and normal individuals, this methodology has the potential to provide a new level of information about disease pathogenesis not previously possible. Two dominant platforms for the construction of high-density microarrays have emerged: cDNA arrays and GeneChips. The first involves robotic spotting of DNA molecules, often derived from EST clone collections, onto a suitable solid phase matrix such as a glass slide. The second involves direct in situ synthesis of sets of gene-specific oligonucleotides on a silicon wafer by an eloquent derivative of the photolithography process. Both cDNA and oligonucleotide arrays are interrogated by hybridization with a fluorescent-labeled cDNA or cRNA representation of the original tissue mRNA. This enables measurement of the expression levels for thousands of mucosal genes in a single experiment. These technologies have recently become less expensive and more widely accessible to all researchers. This review details the principles and methods behind DNA array technology, data analysis and mining, and potential application to research and treatment of IBD. PMID- 11854615 TI - Pancreatic cell lines: a review. AB - Pancreatic cancer has an extremely poor prognosis and lacks early diagnostic and therapeutic possibilities, mainly because of its silent course and explosive fatal outcome. The histogenesis of the disease and early biochemical and genetic alterations surrounding carcinogenesis are still controversial. In vitro studies offer a useful tool to study physiologic, pathophysiologic, differentiation, and transformation processes of cells and to understand some of these shortcomings. The extreme difficulties in isolating individual pancreatic cells and their purification by maintaining their native characteristics have limited research in this area. This review is intended to present and discuss the current availability of rodent and pancreatic cell lines, their differences as well as the difficulties, limitations, and characteristics of these cultured cells. Discussed are in vitro models; ductal, islet, and acinar cell culture; cell differentiation; cell transformation, including genetic and chromosomal alterations; as well as tumor cell markers. Also addressed are the advantages and problems associated with the cell culture in humans and rodents. Advancements in tissue culture technique and molecular biology offer steady progress in this important line of research. The improved methods not only promise the establishment of beta-cell cultures for the treatment of diabetes, but also for studying sequential genetic alterations during pancreatic carcinogenesis and in understanding the tumor cell origin. PMID- 11854616 TI - The gastrin receptor promotes pancreatic growth in transgenic mice. AB - INTRODUCTION: We demonstrated previously, in two different rodent models of pancreatic cancer, that the gastrin receptor is present on malignant pancreatic tumors in spite of the fact that the normal adult rat and mouse pancreas does not express gastrin receptors. AIMS AND METHODOLOGY: To determine whether gastrin receptors mediate pancreatic growth or promote carcinogenesis or both, we created a transgenic mouse that constitutively expresses gastrin receptors in the exocrine pancreas. The transgene construct contained the full-length rat gastrin receptor cDNA sequence under the control of the rat elastase promoter. RESULTS: Receptor presence and function on exocrine pancreatic tissue of transgenic but not control mice were confirmed by (125)I-gastrin-I binding studies and by gastrin stimulation of intracellular calcium release. Eighteen-month-old transgenic animals had larger pancreas-to-body weight ratios than their nontransgenic littermate controls (p < 0.001 for females; p < 0.01 for males); however, histopathologic examination revealed no neoplasms or other abnormalities. CONCLUSION: In both female and male transgenic mice, the expression of the gastrin receptor in the exocrine pancreas is associated with a significant increase in pancreas weight, but it does not appear to promote the development of spontaneous pancreatic tumors. PMID- 11854617 TI - Ligands of peroxisome proliferator-activated receptor-gamma induce apoptosis in AR42J cells. AB - INTRODUCTION: Peroxisome proliferator-activated receptor-gamma (PPAR-gamma) is a ligand-activated transcription factor that controls growth, differentiation, and inflammation in different tissues. Roles of PPAR-gamma activation in pancreatic acinar cells are poorly characterized. AIMS: To examine the effects of PPAR-gamma activation on the induction of apoptosis in rat pancreatic AR42J cells. METHODOLOGY: AR42J cells were treated with ligands of PPAR-gamma, and induction of apoptosis was evaluated by cell viability, DNA-fragmentation, and flow cytometry. RESULTS: Treatment of the cells with ligands of PPAR-gamma (15-deoxy open triangle12,14-prostaglandin J2 or troglitazone) induced apoptosis in a dose dependent manner. Troglitazone-induced apoptosis was not blocked by inhibitors of caspases (acetyl-DEVD-aldehyde and benzoyloxycarbonyl-VAD-fluoromethylketone). Troglitazone induced the expression of pancreatitis-associated protein-1 and clusterin mRNAs. Troglitazone activated c-Jun NH2-terminal kinase/stress activated protein kinase, but inhibited the activation of extracellular signal regulated kinases 1/2. Troglitazone did not activate NF-kappaB, suggesting a role of NF-kappaB-independent pathways. In AR42J cells and isolated pancreatic acini, PPAR-gamma gene and protein were detected. In addition, troglitazone increased the PPAR-dependent transcriptional activity, suggesting that PPAR-gamma is functional in AR42J cells. CONCLUSION: These results indicate that activation of PPAR-gamma induces apoptosis in AR42J cells and imply that PPAR-gamma may be a potential therapeutic target of pancreatic inflammation, because of its anti inflammatory effects in addition to its proapoptotic effects. PMID- 11854618 TI - Expression of a novel zinc-finger cDNA, IA-1, is associated with rat AR42J cells differentiation into insulin-positive cells. AB - INTRODUCTION: IA-1, an insulinoma-associated cDNA-1, encodes a zinc-finger DNA binding protein originally isolated from a human insulinoma subtraction library. AIM: To demonstrate the restriction of IA-1 gene expression in human fetal pancreata of different gestational stages and to determine whether the expression of IA-1 gene is associated with rat AR42J cell differentiation into insulin positive cells. METHODOLOGY: To examine whether the IA-1 gene is associated with pancreatic endocrine cell differentiation, we used a rat pancreatic amphicrine cell line, AR42J, to investigate whether the expression of the IA-1 gene coincides with AR42J cells converting into either endocrine or exocrine lineage. We also examined a set of islet transcription factors that regulate key differentiation steps involved in activating the genes that confer the specialized functions of terminally differentiated pancreatic islet cells. RESULTS: When the AR42J cells were converted into insulin-positive cells induced by GLP-1, insulinoma conditioned-medium, or both, we observed a significant elevated expression of mRNA for IA-1 and islet-specific transcription factors such as Pdx-1, NeuroD/beta2, and Nkx6.1. In contrast, dramatically decreased expression of mRNA for IA-1 and islet-specific transcription factors was displayed when AR42J cells were converted into the acinar-like phenotype by dexamethasone. CONCLUSIONS: IA-1 gene was shown to be developmentally regulated in fetal pancreatic cells, and its expression pattern is consistent with parallel changes in islet-specific transcription factors during the endocrine differentiation of AR42J cells. PMID- 11854619 TI - Superoxide dismutase is induced during rat pancreatic acinar cell injury. AB - INTRODUCTION: Free radicals and their scavengers are supposed to be involved in pancreatitis. AIMS: To investigate the expression of superoxide dismutase (SOD) in rat pancreatic acinar cell injury. METHODOLOGY AND RESULTS: As an in vivo model, male WBN/Kob rats were used. Chronic pancreatitis developed spontaneously at 12 weeks in this model and progressed thereafter, but acinar regeneration was recognized at 20 weeks. By semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR), manganese SOD (MnSOD) mRNA expression peaked at 8 and 20 weeks, whereas copper/zinc SOD (CuZnSOD) mRNA expression peaked at 12 and 20 weeks. Immunohistochemistry confirmed the localization of SOD in acinar cells. Acinar cell apoptosis peaked at 12 and 20 weeks. In an in vitro study, MnSOD mRNA expression peaked at 2 hours after the addition of arginine to culture medium, whereas apoptosis was increased at 24 hours. CONCLUSION: Thus, the induction of SOD around the onset and at the late stage of chronic pancreatitis in the WBN/Kob rats implies pancreatic ischemia and acinar remodeling, respectively. From the in vitro results, MnSOD expression might reflect a defensive mechanism of acinar cells against oxidative stress or pro-apoptotic stimuli. PMID- 11854620 TI - Heterogeneous cell renewal of pancreas in mice: [(3)H]-thymidine autoradiographic investigation. AB - INTRODUCTION: Although cell kinetics of the gastrointestinal mucosa has been extensively examined, that of the pancreas has not been fully analyzed. AIM: To determine the renewal rate of pancreatic cells directly. METHODOLOGY: Postnatal proliferative activity and cellular renewal of the parenchymal cells in ICR mouse pancreas were studied by immunohistochemistry and [(3)H]-thymidine autoradiography. RESULTS: In the single labeling experiments, the proliferative activity of the parenchymal cells in pancreas showed peaks at a few days after birth, decreased thereafter, and reached a low level at 2 months after birth. Continuous labeling experiments revealed that, in the adult pancreas, the half lives of acinar cells, islet cells, and duct epithelial cells were approximately 70 days, 47 days, and 40 days, respectively. Moreover, in the exocrine pancreas, acinar cells of the peri-insular region proliferated more actively than those of the tele-insular region. The renewal rate of glucagon cells was more rapid than that of insulin cells or somatostatin cells. Large ducts showed a high rate of cell renewal in comparison with small ducts. CONCLUSION: The results of this study indicate that cell renewal rates of the pancreas are not homogeneous, but heterogeneous. PMID- 11854621 TI - The role of oxygen-derived free radicals and nitric oxide in cytokine-induced antiproliferation of pancreatic cancer cells. AB - INTRODUCTION: Pancreatic cancer cells are susceptible to antiproliferative effects of cytokines such as tumor necrosis factor-alpha (TNF-alpha). However, little is known about the mechanisms involved. AIM: To determine the mechanisms of the antiproliferative effects of TNF-alpha on pancreatic cancer cells. METHODOLOGY AND RESULTS: In the current study, four of five pancreatic cancer cell lines tested were responsive to the antiproliferative effects of TNF-alpha. In two cell lines, the effects of TNF-alpha were completely abolished by superoxide dismutase, suggesting that superoxide anion mediates the effects. Further, inhibition of nitric oxide (NO) synthase by L-NAME potentiated the TNF alpha response, suggesting a protective role for endogenously produced NO in these two cell lines. MiaPaCa-2 cells, which were unresponsive to the antiproliferative effects of TNF-alpha, produced five times more nitrite than the other cell lines. Treatment of MiaPaCa-2 cells with interferon-gamma (IFN-gamma) increased inducible nitric oxide synthase mRNA as shown by reverse transcription polymerase chain reaction. This induction was potentiated by TNF-alpha and further enhanced by a combination of three cytokines (INF-gamma, TNF-alpha, and interleukin 1-beta (IL-1beta). This combination of cytokines increased nitrite accumulation fourfold and inhibited the proliferation of this resistant cell line. These effects were prevented by L-NAME. CONCLUSIONS: These studies suggest that TNF-alpha inhibits proliferation of pancreatic cancer cells by increasing the production of superoxide anion and that endogenously produced NO protects against this effect. In addition, there is a direct correlation between the amount of NO produced and resistance to TNF-alpha in the only resistant cell line. In contrast, treatment with a combination of IFN-gamma, TNF-alpha and IL 1beta upregulates inducible nitric oxide synthase, and the resulting markedly enhanced NO production inhibits pancreatic cancer cell growth. This pathway may provide a valuable target for therapy of this devastating disease. PMID- 11854622 TI - Histologic features of venous invasion, expression of vascular endothelial growth factor and matrix metalloproteinase-2 and matrix metalloproteinase-9, and the relation with liver metastasis in pancreatic cancer. AB - INTRODUCTION: Pancreatic cancer frequently is associated with venous invasion and hematogenous metastasis. AIMS: To determine morphologic features of invaded veins, intratumoral vascular composition, the correlation with liver metastasis, and expression of vascular endothelial growth factor (VEGF), matrix metalloproteinase (MMP)-2 and MMP-9, and the mechanism of development of hematogenous metastasis. METHODOLOGY: We examined 32 patients with resected pancreatic cancer: 18 had postoperative liver metastasis, and 14 had no liver metastasis. Specimens were examined to determine the composition of veins and microvessels by staining of victoria-blue and CD34. We also investigated expression of VEGF, MMP-2, and MMP-9 by immunohistochemical staining. RESULTS: Venous invasion was detected in 31 of 32 patients. Invaded venous densities of middle- and large-sized veins were significantly higher in patients with liver metastasis than in those with nonliver metastasis, and they were related to MMP-2 and MMP-9 overexpression. Invaded veins with fragmentation of the lumen through cancer cells were considered to be an intravasation of cancer (destroyed type vein), and their numbers were significantly related to liver metastasis, and MMP 2 and MMP-9 overexpression. CONCLUSION: In conclusion, almost all the patients with pancreatic cancer showed venous invasion, indicating that invasion into large veins and destroyed type veins could be a risk factor for liver metastasis and that increased expression MMP-2 and MMP-9 were related to such invasion. PMID- 11854623 TI - Acetylcholine-induced zymogen granule exocytosis: comparison between acini and single pancreatic acinar cells. AB - INTRODUCTION: Numerous studies have been carried out on the agonist-evoked calcium responses of single pancreatic acinar cells; however, several reports have shown that dissociation of the exocrine pancreas into predominantly single cells has an adverse effect on agonist-evoked amylase secretion. AIMS AND METHODOLOGY: To determine whether single acinar cells behave in an anomalous manner compared with cells within an intact acinus, we measured exocytosis in both single acinar cells and acini (2-5 cells) present in the same preparation. Exocytosis of individual zymogen granules was quantified in real-time by using the technique of continuous time-differential analysis of brightfield digital images. RESULTS: Basal rates of exocytosis were low in both single cells and intact acini. Application of 10 microM acetylcholine for 6 minutes stimulated a biphasic secretory response in acinar cells. Additionally, we found that exocytotic events occur repetitively in specific locations within the apical domain; i.e., there are exocytotic "hot spots." There were no statistically significant differences between the exocytotic rates, nor were there any differences in the characteristics of the exocytotic hot spots of single cells compared with those of acini. CONCLUSION: We conclude that time-differential analysis of brightfield images appears to be a useful tool for the investigation of the role of gap junctions in zymogen granule exocytosis and that single acinar cells provide a reasonable model for studies of acinar cell signaling and secretion. PMID- 11854624 TI - Thiazolidinedione derivatives as novel therapeutic agents to prevent the development of chronic pancreatitis. AB - INTRODUCTION: Thiazolidinedione derivatives are known to be novel insulin sensitizing agents and ligands of a nuclear hormone receptor peroxisome proliferator-activated receptor gamma (PPARgamma). Recently, ligands of PPARgamma have been shown to modulate proinflammatory cytokine production and NF-kappaB activation. AIM: To show that thiazolidinedione derivatives interfere with the development of chronic pancreatitis. METHODOLOGY: Rat chow containing 0.2% troglitazone was administered from 1 month to 7 months of age in WBN/Kob rats with spontaneous chronic pancreatitis. Morphologic evaluation of the pancreas was performed at 4 months and 7 months of age. Pancreas weight, protein, amylase, and insulin contents also were determined. Changes of cytokine levels were detected by enzyme-linked immunosorbent assay or semiquantitative reverse transcription polymerase chain reaction. Localization and expression of PPARgamma in the pancreas and isolated peritoneal macrophages were examined by immunohistochemical study. RESULTS: Administration of troglitazone reduced the severity of morphologic pancreatic damage including inflammatory cell infiltration, and fibrosis markedly improved by the administration of troglitazone. Further, troglitazone was able to prevent the decrease in amylase content and pancreas atrophy that were observed in WBN/Kob rats. Serum IL-8 levels and TNF-alpha mRNA levels in the pancreas were significantly elevated in WBN/Kob rats, and these were dramatically attenuated by troglitazone. Peritoneal macrophages isolated from normal rats expressed PPARgamma at low levels, whereas those from WBN/Kob rat abundantly expressed PPARgamma. CONCLUSION: Troglitazone prevented the progression of pancreatic inflammatory process in an animal model of chronic pancreatitis. Macrophages may be one of the targets of the PPARgamma ligand to attenuate the severity of chronic pancreatitis, partially mediated by the inhibition of proinflammatory cytokine gene expression. PMID- 11854625 TI - Influence of contrast-enhanced computed tomography on course and outcome in patients with acute pancreatitis. AB - INTRODUCTION: Many of the complications in severe acute pancreatitis result from the amplifying effects of microcirculatory disruption. Contrast medium may cause significant additional reductions of capillary flow, which has been shown to aggravate acute pancreatitis in experimental studies. AIM: To investigate the role of serial contrast-enhanced computed tomography (CECT) in patients with acute pancreatitis. METHODOLOGY: A retrospective analysis evaluated 302 patients with moderate to severe acute pancreatitis. Among these patients, 264 underwent CECT within 96 hours of the onset of symptoms and again during the course, but in 38 patients no serial CECT was performed. Outcome measurement was analyzed by comparison of hospital stay and mortality rate between the two patient groups. Influences of contrast medium on severity of disease were detected by monitoring complications during the course of treatment, C-reactive protein, and APACHE II score. RESULTS: The 1-month mortality rate was less in patients with CECT (6.4% versus 15.8%, p <0.05). There were no significant differences considering the incidence of additional complications, and hospital stay was not significantly longer (29 +/- 36 versus 19 +/- 13 days). C-reactive protein and APACHE II score had similar time courses. CONCLUSION: Contrast-enhanced computed tomography remains crucial in identifying patients with acute pancreatitis at high risk to develop necrosis of the pancreas and systemic complications. Contrast medium has been found to aggravate acute pancreatitis in animal models. As compared with the patient group without being exposed to contrast medium, however, this study did not show a deterioration of acute pancreatitis by administration of contrast medium in men. PMID- 11854626 TI - Induction of chemokines in rat pancreatic acinar cell injury. AB - INTRODUCTION: The pathogenesis of chronic pancreatitis (CP), especially of acinar cell injury, is still unclear. Interleukin (IL)-8 is a chemokine that is involved in various inflammatory diseases. AIM: To examine whether IL-8 and other chemokines are expressed in experimental acinar cell injury. METHODOLOGY: IL-8 expression was analyzed in spontaneous CP in the WBN/Kob rat and in rat pancreatic acinar AR4-2J cells treated with various stimuli using reverse transcription-polymerase chain reaction (semiquantitative) and immunohistochemistry. RESULTS: Chronic pancreatitis developed at 12 weeks in the WBN/Kob rats. IL-8, macrophage chemoattractant protein-1, and macrophage inflammatory protein-2 mRNA was expressed from 4 weeks and peaked at 12 weeks. Immunohistochemistry showed a strong expression of IL-8 in acinar cells, proliferating ductular cells, and interstitial infiltrating cells. In contrast, normal pancreatic tissues lacked IL-8 expression. Further, IL-8 mRNA and protein were detectable in AR4-2J cells treated with the various stimuli, such as menadione, tumor necrosis factor-alpha and transforming growth factor beta1. CONCLUSION: These results suggest that IL-8 is expressed in the pancreatic parenchyma and infiltrates in CP and that it plays a role in the initial pathogenesis of CP together with other chemokines and cytokines. PMID- 11854627 TI - Metabolism of glucose by rat pancreatic islets and kidney: comparison of membrane preparations with intact tissue. AB - INTRODUCTION: Insulin secretion in the pancreatic islet of Langerhans is closely related to glucose oxidation. Our previous membrane preparations from the islet of Langerhans and kidney contained Na+/K+ ATPase. Although ATP is a modulator of insulin release, we have not demonstrated that membranes contain glycolytic enzymes. AIMS: To compare glucose oxidation in intact islets of Langerhans with that seen in islet membranes, and to examine renal tissue because of easy access to this tissue and because our previous work contrasted islets and kidney Na+/K+ ATPase activity. METHODOLOGY: To examine whether membrane preparations of both islets and kidney contain glycolytic activity, we designed an apparatus that determines tissue glucose oxidation rates by measuring 14CO2 production from 14C 1-D-glucose. In contrast to previous methods, O2 was supplied and pH was maintained in physiologic ranges throughout the incubations. RESULTS: At 5.5 mM glucose, oxidation was linear over 2 hours in intact islets and kidney. Islets produced approximately twice the 14CO2 as kidney, or 1000 nmol CO2/mg protein/hour. Membranes oxidized glucose at less than 10% of the rates observed with intact tissue. Concentration dependency was linear from 0-11.0 mM glucose; however, at 16.5 mM, oxidation decreased to less than half of that seen at 11.0 mM. CONCLUSION: Our "membrane" preparations contain some glycolytic activity but are unlikely to contribute significant amounts of ATP in most studies of ATP hydrolysis, such as that seen in experiments that measure ATPase activity. PMID- 11854628 TI - Intrasurgical pancreas cytology. AB - INTRODUCTION: A differential therapy of chronic pancreatitis and carcinoma calls for evaluation of the validity of findings. Presurgical suspicion of a carcinoma often requires intrasurgical diagnostics such as excisional biopsies, punch biopsies, and fine-needle aspiration cytology (FNAC) for confirmation. AIMS: To evaluate FNAC as an intraoperative diagnostic method of very high probability. METHODOLOGY: Intrasurgical fine-needle aspiration biopsy and cytologic assessment were carried out in 474 patients. The indications for operative therapy and FNAC were suspicion of pancreatic tumor, chronic pancreatitis even without suspicion of tumor, and pathologic alterations found during other surgeries in the upper abdomen. RESULTS: The level of sensitivity was 93.1%, specificity was 99.1%, predictive value of positive results was 99.2% and of negative results was 92.1%. CONCLUSION: FNAC is a suitable method for intrasurgical confirmation of pancreatic carcinoma. It can be performed safely, effectively, and rapidly. PMID- 11854629 TI - Efficacy of hormone replacement therapy assessed by quantitative ultrasonography. PMID- 11854631 TI - The significance of psammoma bodies that are found incidentally during endometrial biopsy. AB - OBJECTIVE: The purpose of this study was to elucidate the significance of psammoma bodies that are found incidentally during endometrial biopsy. STUDY DESIGN: We reviewed the medical records of 11 women who were found to have psammoma bodies during endometrial biopsy over an 18-month period and extracted data that included demographic information, extent of evaluation, and pathologic findings. RESULTS: Ten women were postmenopausal and underwent endometrial biopsy for abnormal uterine bleeding while using combined hormone replacement. Most women underwent dilation and curettage with hysteroscopy plus either laparoscopy or ultrasonography. Notable findings included: 5 women with endometrial polyps, 2 women with endometriosis, 1 woman with endosalpingiosis, and 1 woman with a mature cystic teratoma. No endometrial or adnexal malignancies were identified. CONCLUSION: This series represents the largest series to date regarding psammoma bodies that have been found incidentally during endometrial biopsy. All psammoma bodies were associated with benign findings. Further evaluation of the endometrial cavity with hysteroscopy or sonohysterography, plus some form of adnexal assessment, is a reasonable definitive evaluation scheme. PMID- 11854630 TI - Expression of receptors for luteinizing hormone-releasing hormone in human ovarian and endometrial cancers: frequency, autoregulation, and correlation with direct antiproliferative activity of luteinizing hormone-releasing hormone analogues. AB - OBJECTIVE: Several recent reports have demonstrated the expression of luteinizing hormone-releasing hormone receptors by human ovarian and endometrial cancers. Controversy persists on the relevance of this finding, in particular whether these receptors mediate direct antiproliferative effects of luteinizing hormone releasing hormone analogues. We correlated the expression of luteinizing hormone releasing hormone receptors by well-characterized ovarian and endometrial cancer cell lines with the ability of luteinizing hormone-releasing hormone analogues to reduce their proliferation and studied the autoregulation of luteinizing hormone releasing hormone receptor expression by luteinizing hormone-releasing hormone agonist triptorelin and antagonist cetrorelix. The expression of luteinizing hormone-releasing hormone receptors was assessed in a series of specimens from primary ovarian and endometrial cancers. STUDY DESIGN: Luteinizing hormone releasing hormone receptor expression was assessed by semiquantitative reverse transcriptase-polymerase chain reaction and radioligand binding assay. Antiproliferative effects were ascertained by proliferation assays in the absence or presence of luteinizing hormone-releasing hormone analogues. RESULTS: Ovarian (4/6 cell lines) and endometrial (5/6 cell lines) cancer cell lines expressed luteinizing hormone-releasing hormone receptors. The proliferation of these luteinizing hormone-releasing hormone receptor-positive cell lines was dose- and time-dependently reduced by agonistic and antagonistic luteinizing hormone releasing hormone analogues. Luteinizing hormone-releasing hormone receptor density was reduced to 80% of controls (control, 100 %; P <.001) by luteinizing hormone-releasing hormone analogues. Seventy percent of primary ovarian cancers and 83% of primary endometrial cancers expressed luteinizing hormone-releasing hormone receptors. CONCLUSION: These findings suggest that luteinizing hormone releasing hormone receptors that are expressed by human ovarian and endometrial cancer cell lines mediate direct antiproliferative effects of luteinizing hormone releasing hormone analogues. Because most respective primary cancers expressed luteinizing hormone-releasing hormone receptors, these receptors might be used for novel antiproliferative therapeutic approaches and should be further evaluated. PMID- 11854632 TI - Growth of the uterus and mammary glands and vaginal cytologic features in extremely premature infants with postnatal replacement of estradiol and progesterone. AB - OBJECTIVE: Growth of the uterus and the mammary glands and changes in vaginal cytologic features are known to be estrogen dependent and were evaluated to proof the biologic effectiveness of a postnatal replacement of estradiol and progesterone in extremely premature infants. STUDY DESIGN: Thirty female infants with a mean gestational age of 26.4 weeks (24.1-28.7 weeks) and a mean birth weight of 708 g (370-990 g) were investigated. Fifteen infants received postnatal replacement of estradiol and progesterone for 6 weeks to maintain intrauterine plasma levels of estradiol and progesterone. Uterine size and the diameter of mammary glands were assessed repeatedly by ultrasound scans and palpation. Vaginal smears were also obtained. RESULTS: The uterus and mammary glands showed significant growth during the hormone replacement, but growth was not observed in nontreated infants. Vaginal smears showed high karyopyknotic and eosinophilic indices in both groups at birth; the indices remained significantly higher at 3 and 6 weeks in the hormone-treated infants. CONCLUSION: The biologic effectiveness of postnatal estradiol and progesterone replacement in extremely premature infants was proved. PMID- 11854633 TI - Arterial stiffness is not improved in long-term use of estrogen. AB - OBJECTIVE: The purpose of this study was to compare arterial stiffness in long term users of estrogen who were postmenopausal, age-matched women who did not use estrogen, and women of fertile age. STUDY DESIGN: In this clinical cross sectional study, carotid, femoral, and aortic stiffness (estimated as elastic modulus and stiffness beta index) were assessed by ultrasonic phase-locked echo tracking in 17 women who were postmenopausal and who were treated with 17 beta estradiol implants (mean age, 68.8 years; mean duration of treatment, 20 years), 17 age-matched (+/-1 year) untreated women, and 20 women in the fertile age period. RESULTS: Compared with women of fertile age, both postmenopausal groups had significantly higher stiffness elastic modulus and stiffness beta index (carotid, P <.001; femoral, P <.05; aorta, P <.001). However, for all 3 arteries, both stiffness indices were similar in estrogen users and nonusers and did not differ significantly. These results remained after an adjustment for systolic blood pressure, body mass index, and low-density lipoprotein cholesterol levels. CONCLUSION: These data indicate that the arterial stiffening that is related to aging/menopause is not substantially affected by long-term estrogen therapy. PMID- 11854634 TI - Oral contraceptives and the risk of focal nodular hyperplasia of the liver: a case-control study. AB - OBJECTIVE: A role of female hormones, including oral contraceptives, has been suggested in the etiology of focal nodular hyperplasia of the liver. There is, however, no epidemiologic quantification of this relationship. STUDY DESIGN: A hospital-based case-control study was conducted in Italy of 23 women with histologically confirmed focal nodular hyperplasia of the liver and 94 controls in the hospital for acute diseases. Odds ratios (ORs) were computed by use of multiple logistic regression models. RESULTS: Focal nodular hyperplasia was not associated with menstrual and reproductive factors. Ever oral contraceptive use was reported by 83% of cases and 59% of controls. The multivariate OR was 2.8 (95% confidence interval [CI], 0.8-9.4) for ever use, and 4.5 (95% CI, 1.2-16.9) for use > or = 3 years. The trend in risk with duration was statistically significant. CONCLUSIONS: This study confirms previous clinical observations, and provides a quantitative estimate of the association between use of oral contraceptives and focal nodular hyperplasia of the liver. PMID- 11854635 TI - Pulmonary embolism and stroke in relation to pregnancy: how can high-risk women be identified? AB - OBJECTIVE: Risks of circulatory diseases are increased substantially during late pregnancy and around the time of delivery. This study was undertaken to determine whether preeclampsia, multiple pregnancy, or cesarean delivery account for the majority of pregnancy-related risks of pulmonary embolism and stroke (caused by hemorrhage, infarction, and intracranial venous thrombosis). STUDY DESIGN: We analyzed a population-based cohort of 1,003,489 deliveries in Sweden. Relative risks of pulmonary embolism and stroke were modeled by use of Poisson regression. RESULTS: Preeclampsia was associated with 3- to 12-fold increases in risks of pulmonary embolism and stroke during late pregnancy, at delivery, and in the puerperium, and similar increases in risks were also observed for multiple pregnancies and cesarean delivery. These strong associations could not explain the overall pregnancy-related risks of pulmonary embolism and stroke. CONCLUSION: Preeclampsia, multiple birth, and cesarean delivery are important risk factors for pulmonary embolism and stroke, but they do not account for the majority of the excess risks associated with pregnancy. PMID- 11854636 TI - Does insulin secretion in patients with one abnormal glucose tolerance test value mimic gestational diabetes mellitus? AB - OBJECTIVE: The purpose of this study was to investigate the insulin response to a 3-hour oral glucose tolerance test and to compare the insulin levels in the gestational diabetes mellitus and single abnormal test value groups with a nondiabetic control group. STUDY DESIGN: One hundred ten Turkish women with uncomplicated pregnancy participated in this prospective controlled study between 24 to 28 weeks of gestation. A 100-g 3-hour oral glucose tolerance test was given, and glucose and insulin plasma levels were assayed. The subjects were classified according to established criteria. Early-phase insulin secretion was assessed by the insulinogenic index. Total insulin secretion was assessed by mean insulin level during the oral glucose tolerance test; insulin resistance was assessed by fasting insulin concentration and by the use of the homeostasis model. Data were analyzed by the Student t test and 1-way analysis of variance, with posthoc Bonferroni correction. RESULTS: The fasting insulin levels of patients with normal oral glucose tolerance test results were significantly lower than those of patients with gestational diabetes mellitus and a single value abnormality (P <.001 and P <.005, respectively). The insulinogenic index as a marker of early-phase insulin secretion was significantly lower in gestational diabetes mellitus, compared with that of patients with normal oral glucose tolerance test results (P <.05). The worsening of glycemic profile from normal oral glucose tolerance test results to gestational diabetes mellitus was associated with an increase in the homeostasis model; no significant difference was found between gestational diabetes mellitus and a single value abnormality group in terms of both the homeostasis model and the insulinogenic index. Values for total insulin secretion were highest in gestational diabetes mellitus, followed by the single value abnormality group, both significantly differing from the values of patients with normal oral glucose tolerance test results (P <.001 and P <.005, respectively). CONCLUSION: In this prospective study of Turkish subjects, we found a striking similarity in terms of patient characteristics between the gestational diabetes mellitus group and the single value abnormality group. Additionally, when we used fasting insulin level and insulin resistance as 2 separate criteria of analysis, patients with single value abnormality were indistinguishable from patients with gestational diabetes mellitus; both groups were significantly different from the normal oral glucose tolerance test group. Our findings suggest that a single abnormal test value on an oral glucose tolerance test should be regarded as a pathologic finding and that the patient with a single abnormal test value may be treated similarly to the patient with gestational diabetes mellitus. PMID- 11854637 TI - Syncytin, a novel human endogenous retroviral gene in human placenta: evidence for its dysregulation in preeclampsia and HELLP syndrome. AB - OBJECTIVE: A novel human endogenous retroviral element, designated as syncytin, has been suggested as a contributor to normal placental architecture, especially in the fusion processes of cytotrophoblasts to syncytiotrophoblasts. We tested the hypothesis of whether the gene expression of syncytin may be altered in cases with placental dysfunction such as preeclampsia or HELLP (hemolysis, elevated liver enzymes, low platelets) syndrome. STUDY DESIGN: We included 30 women with normal pregnancies, 16 with preeclampsia, and 6 with HELLP syndrome. After delivery, messenger ribonucleic acids (mRNA) of syncytin, glyceraldehyde-3 phosphate dehydrogenase and beta-actin were analyzed in placental villi with use of quantitative real-time polymerase chain reaction. RESULTS: In placental villi, syncytin mRNA/beta-actin mRNA and syncytin mRNA/glyceraldehyde-3-phosphate dehydrogenase mRNA ratios were lower in patients with preeclampsia (P <.05) or HELLP syndrome than in healthy control subjects. CONCLUSION: A reduced placental expression of syncytin may contribute to altered cell fusion processes in placentogenesis and disturbed placental function in hypertensive disorders of pregnancy. PMID- 11854638 TI - Endothelial junctional protein redistribution and increased monolayer permeability in human umbilical vein endothelial cells isolated during preeclampsia. AB - OBJECTIVE: To examine monolayer permeability and junctional protein distribution and expression in endothelial cells isolated from women with preeclampsia and from women with normal pregnancies. We hypothesized that increased endothelial monolayer permeability in preeclampsia reflects altered monolayer barrier properties produced by disorganization of endothelial cell junction proteins. STUDY DESIGN: Endothelial cells were isolated from umbilical veins from women with normal pregnancies (n = 9) and from women with preeclampsia (n = 9) immediately after delivery. In the first passage of endothelial cells, the permeability was determined by measurement of horseradish peroxidase passage through confluent cell monolayers grown on transwell filters, and the distribution and protein expression of vascular endothelial cadherin and occludin were evaluated by use of immunofluorescent staining of the proteins and Western blot analysis. The distribution of vascular endothelial cadherin was also evaluated in the second and third passage endothelial cells. Messenger ribonucleic acid expression of vascular endothelial cadherin and occludin were examined by reverse transcriptase polymerase chain reaction. Data were expressed as mean values (+/- SE) and were analyzed by use of an unpaired t test. RESULTS: The relative monolayer permeability was significantly increased in the endothelial cells isolated from women with preeclampsia compared with those isolated from women with normal pregnancies (0.271 +/- 0.049 versus 0.086 +/- 0.031 for DeltaOD(470); P <.05). Vascular endothelial cadherin expression for normal endothelial cells showed a continuous staining of the junctional protein that surrounded cell borders. In comparison, vascular endothelial cadherin in endothelial cells from preeclamptic pregnancies exhibited disorganized staining and vascular endothelial cadherin fibrils were retracted, with gaps present at the cell borders. The expression of occludin showed a pattern similar to that of vascular endothelial cadherin in both normal and preeclamptic conditions. Western blot results for expression of vascular endothelial cadherin and occludin also showed decreased expression of junctional proteins. The altered endothelial cell junctional protein distribution and expression of vascular endothelial cadherin and occludin observed in the first passage of endothelial cells from preeclamptic pregnancies was restored to normal by the time cells reached the third passage in vitro. There was no statistical difference in mRNA expression for the vascular endothelial cadherin and occludin between normal endothelial cells and those from preeclamptic pregnancies. CONCLUSION: In preeclampsia, increased endothelial cell monolayer permeability appears to reflect disorganized and diminished expression of endothelial cell junctional proteins. The latter response is mediated at the posttranscriptional level. These findings provide new insights about the cellular and molecular basis for altered endothelial cell integrity and barrier dysfunction that are associated with preeclampsia. PMID- 11854639 TI - The cost-effectiveness of routine versus restrictive episiotomy in Argentina. AB - OBJECTIVE: This article provides the estimates of the cost implications of switching from routine to restrictive episiotomy in 2 provinces in Argentina (Santa Fe and Salta) from the viewpoint of the health provider. STUDY DESIGN: A decision-tree model was constructed that used the probabilities and patient outcomes (the results of a trial in Argentina), resource use, cost, and local epidemiologic data from interviews with obstetricians in the selected provinces and from literature reviews. Probabilistic sensitivity analysis was conducted, which provided 90% confidence ranges for the cost data. RESULTS: For each low risk vaginal delivery, there is a potential reduction in provider cost of $20.21 (range, $19.36-$21.09) with a restrictive policy of episiotomy in Santa Fe province and a reduction of $11.63 (range, $10.89-$12.42) in Salta province. CONCLUSION: The more effective policy of restrictive episiotomy is also less costly than that of routine episiotomy. The results are robust and consistent in both provinces. Further research is required to confirm the appropriate indications for episiotomy and the impact on outcomes of variations in episiotomy cost rates. PMID- 11854640 TI - Buccal versus intravaginal misoprostol administration for cervical ripening. AB - OBJECTIVE: The purpose of this study was to compare the efficacy of misoprostol that is administered in the buccal pouch with the intravaginal route of administration. STUDY DESIGN: One hundred fifty-seven pregnant women with a singleton live gestation, Bishop score of <7, estimated fetal weight of <4500 g, and gestational age of >24 weeks were randomized to receive misoprostol that would be placed either in the buccal pouch or vagina every 6 hours. In the buccal group, after the first 2 doses of 200 microg, the dose was increased to 300 microg for the duration of the study (up to a total of 1600 microg). In the vaginal group, after the first 2 doses of 50 microg, the dose was increased to 100 microg for the duration of the study (up to a total of 500 microg). The primary outcome variable was the interval from the first dose to vaginal delivery. Power calculations indicated the need to enroll 71 patients in each arm of the study, which would allow for the detection of a 4-hour reduction in vaginal birth interval for buccal misoprostol. RESULTS: The hours from drug administration to vaginal delivery were similar between the buccal and vaginal groups (23.5 +/- 20.8 hours versus 21.3 +/- 13.4 hours), respectively. Thirty five women (63%) versus 34 women (67%) delivered vaginally within 24 hours (P = not significant). The incidence of tachysystole was higher in the buccal group, 28 occurrences (38%) versus 15 occurrences (19%; P =.01). CONCLUSION: Buccal misoprostol is effective for cervical ripening but results in a higher incidence of tachysystole than does intravaginal administration. PMID- 11854641 TI - Chronic intrauterine infection and inflammation in the preterm rabbit, despite antibiotic therapy. AB - OBJECTIVE: In a pregnant rabbit model using intracervical inoculation of Escherichia coli with delayed antibiotic therapy, we investigated the rate of positive cultures and histologic inflammation of maternal and fetal compartments and the concentration of tumor necrosis factor-alpha in the amniotic fluid for up to 5 days. STUDY DESIGN: New Zealand White rabbits at 70% gestation were inoculated intracervically with 10(3) - 10(4) colony-forming units of E coli per uterine horn. At varying intervals after inoculation (0.5 - 4.0 hours), antibiotic therapy was initiated with ampicillin-sulbactam. Primary outcomes were positive cultures and histologic inflammation score. Tumor necrosis factor-alpha levels in the amniotic fluid were determined by bioassay. RESULTS: A total of 60 animals were inoculated with E coli. At the endpoint, uterine cultures were positive more commonly than in the fetus or amniotic fluid (41.8% vs 27.5% vs 17.3%, respectively), which was consistent with an ascending pathway of infection. Inflammation scores were similar in uterus and placenta but lower in fetal lung and absent in fetal brain (2.8 vs 3.1 vs 0.84 vs 0.0, respectively). Comparing the durations of delay in antibiotic administration, we found a significant increase in positive uterine cultures and a significant increase in histologic inflammation score with increasing delay. The proportion of dead pups within a litter was significantly associated with the log of the tumor necrosis factor-alpha concentration in amniotic fluid and the degree of histologic inflammation in the uterus, but not with amniotic fluid or other culture positivity. CONCLUSION: The administration of therapeutic doses of antibiotic does not consistently eradicate bacteria from the rabbit uterus nor, more importantly, from the fetus and the amniotic fluid. Obtaining a negative amniotic fluid culture does not exclude either infection in the decidua or the fetus or histologic inflammation with tumor necrosis factor-alpha elaboration. PMID- 11854642 TI - Outcome after elective labor induction in nulliparous women: a matched cohort study. AB - OBJECTIVE: To determine whether elective induction of labor in nulliparous women is associated with changes in fetomaternal outcome when compared with labor of spontaneous onset. STUDY DESIGN: All 80 labor wards in Flanders (Northern Belgium) comprised a matched cohort study. From 1996 through 1997, 7683 women with elective induced labor and 7683 women with spontaneous labor were selected according to the following criteria: nulliparity, singleton pregnancy, cephalic presentation, gestational age at the time of delivery of 266 to 287 days, and birth weight between 3000 and 4000 g. Each woman with induced labor and the corresponding woman with spontaneous labor came from the same labor ward, and they had babies of the same sex. Both groups were compared with respect to the incidence of cesarean delivery or instrument delivery and the incidence of transfer to the neonatal ward. RESULTS: Cesarean delivery (9.9% vs 6.5%), instrumental delivery (31.6% vs 29.1%), epidural analgesia (80% vs 58%), and transfer of the baby to the neonatal ward (10.7% vs 9.4%) were significantly more common (P <.01) when labor was induced electively. The difference in cesarean delivery was due to significantly more first-stage dystocia in the induced group. The difference in neonatal admission could be attributed to a higher admission rate for maternal convenience when the women had a cesarean delivery. CONCLUSION: When compared with labor of spontaneous onset, elective labor induction in nulliparous women is associated with significantly more operative deliveries. Nulliparous women should be informed about this before they submit to elective induction. PMID- 11854643 TI - Direct prediction of amniotic fluid volume in the third trimester by 3 dimensional measurements of intrauterine pockets: a tool for routine clinical use. AB - OBJECTIVE: To develop an ultrasonographic technique for direct prediction of amniotic fluid volume. STUDY DESIGN: While women with near-term pregnancies were in the supine position, the intrauterine space was divided into 2 hemispheres and each hemisphere was further subdivided into 3 segments. The maximum distances along the 3 axes of amniotic fluid pocket in each segment were measured, and the volume was calculated as a half-ellipsoid or a crescent-cylinder. A sum of these calculations (prediction of amniotic fluid volume [pAFV]) was made and correlated with amniotic fluid volume measured at delivery (mAFV). RESULTS: In 714 deliveries, a high correlation coefficient of 0.896 between mAFV (mean +/- SD, 296.5 +/- 199.2 mL; range, 20 to 1480 mL) and pAFV (mean +/- SD, 300.2 +/- 180.8 mL; range, 27 to 1262 mL) was obtained. The regression equation was mAFV = 0.968pAFV + 8.68. CONCLUSION: The regression equation suggested that pAFV could be a direct predictor of intrauterine amniotic fluid volume in the third trimester. PMID- 11854644 TI - Adolescents' ambivalence about becoming pregnant predicts infrequent contraceptive use: a prospective analysis of nonpregnant African American females. AB - This study prospectively investigated the hypothesis that ambivalence about becoming pregnant is associated with comparatively less frequent use of contraception. Findings from a sample of sexually active African American adolescent, nonpregnant females (n = 375) supported this hypothesis. Prevention programs may benefit from content designed to redress adolescents' positive beliefs about early parenthood. PMID- 11854645 TI - Acute renal failure complicating severe preeclampsia requiring admission to an obstetric intensive care unit. AB - OBJECTIVE: To determine risk factors and outcomes for women with severe preeclampsia and renal failure. STUDY DESIGN: Retrospective study from 1995 to 1998 of all women with renal failure who were admitted to the obstetric intensive care unit at Groote Schuur Hospital, South Africa. A total of 89 women were identified with severe preeclampsia defined as blood pressure > or = 160/110 mm Hg and > or = 2+ proteinuria, renal failure defined as a creatinine level of > or = 1.13 mg/dL, and oliguria defined as < 100 mL urine produced in 4 hours; 72 charts were available for analysis. A comparison was made between the 3 groups, which were defined by the maximum recorded creatinine levels. RESULTS: Of the 72 women, 31 women (43%) were primiparous and 41 (57%) were multiparous. Median gestation at delivery was 32 weeks (range, 21-40 weeks). The median maximum creatinine was 3.85 mg/dL (range, 1.13-12.50 mg/dL). Twelve women (16%) had a history of chronic renal disease or hypertension, and 36 women (50%) had HELLP syndrome and 23 (32%) abruptio placentae. All women with severe renal impairment had either abruptio placentae or hemolysis, elevated liver enzymes, and low platelet count (HELLP) syndrome. Perinatal mortality was 38% (27/72). However, in this series only 7 women (10%) required dialysis in the short term and none required long-term dialysis or kidney transplant. There were no maternal deaths. CONCLUSIONS: In women with severe preeclampsia and renal failure, major obstetric complications were common and perinatal outcome was poor. However, the need for dialysis was infrequent, with only 10% women requiring transient dialysis, and there were no cases of chronic renal failure that required dialysis or kidney transplant. PMID- 11854646 TI - Maternal plasma corticotropin-releasing hormone trajectories vary depending on the cause of preterm delivery. AB - OBJECTIVE: Although second trimester maternal plasma corticotropin-releasing hormone concentrations are elevated in women who are destined to deliver preterm, the sensitivity and positive predictive value of an individual test of corticotropin-releasing hormone concentration is low. This poor screening performance may be due in part to the observation that the causes of preterm delivery are heterogenous, with potentially different effects on corticotropin releasing hormone production. We have reanalyzed data from a previous cohort of women with multiple samplings to determine whether the trajectory of increase of placental corticotropin-releasing hormone provided more information than did a single sample. STUDY DESIGN: In this cohort, where 2 to 4 samples that had been assayed for corticotropin-releasing hormone were available on 305 women, the general form of the exponential equation y = ae(bt) was fitted to the corticotropin-releasing hormone data of each individual by the Gauss-Newton nonlinear least squares method, which generated the parameters, y-intercept and rate of rise, in corticotropin-releasing hormone concentration for each woman. Nonparametric statistical techniques, including bootstrapping, were applied to compare the results for the group of women who delivered at term with the group of women who delivered spontaneously preterm and the group of women who were delivered preterm by induction or cesarean delivery because of obstetric indication. RESULTS: The 3 clinically defined groups have significantly different parameters, y-intercept and rate of rise, which describe the corticotropin releasing hormone trajectory. The group that delivered preterm because of obstetric indications had a similar mean y-intercept but significantly greater mean rate of rise in corticotropin-releasing hormone concentration than the term group (0.3491 vs 0.1788; 95% CI, 0.2331-0.4615 vs 0.1394-0.2330). The group that delivered spontaneously preterm had a significantly greater mean y-intercept than the group that delivered preterm because of obstetric indication (17.08 vs 1.83; 95% CI, 5.89-28.43 vs 0.03-5.19) but a similar mean rate of rise to the group that delivered at term. CONCLUSION: These data suggest that spontaneous preterm delivery is associated with an abnormal setting of the production of corticotropin-releasing hormone that is present from very early in pregnancy, although women who experience an induced preterm delivery are characterized by rapidly rising placental corticotropin-releasing hormone concentrations. These data further suggest that clinical abnormalities that are associated with preterm delivery by induction or cesarean delivery are associated with abnormalities that lead to a rapidly increasing corticotropin-releasing hormone concentrations. Trajectories for corticotropin-releasing hormone provide information beyond that obtained from a single sample. PMID- 11854647 TI - Effects of chronic hypoxia and protein malnutrition on growth in the developing chick. AB - OBJECTIVE: The purpose of this study was to determine how chronic hypoxia and/or protein malnutrition in ovo affect growth in developing chicks. STUDY DESIGN: Chicken eggs were incubated under normoxic (21% oxygen; n = 30 eggs) or hypoxic (14% oxygen; n = 80 eggs) conditions. Hypoxia was imposed from day 0 (n = 38 eggs), day 10 (n = 22 eggs), or from day 0 to 10 (n = 20 eggs). Protein malnutrition alone (n = 20 eggs) or in combination with hypoxia (n = 24 eggs) was induced by removal of 10% of the estimated total albumin content of the egg. Embryos/chicks were killed and weighed at day 10, 15, or immediately after hatch; organs were removed and weighed. RESULTS: Embryos to which hypoxia was imposed from day 0 weighed less than control embryos at day 10, which stayed the same until hatch (64.67% +/- 3.56% egg mass vs 69.36% +/- 3.90% [mean +/- SD]; P <.05). Malnourished chicks at day 15 and at hatch (63.42% +/- 4.28%; P <.05) weighed less than control chicks, as did malnourished plus hypoxia chicks (59.74% +/- 3.41%; P <.001). Malnourished plus hypoxia chicks weighed less than malnourished chicks alone (P <.05). Embryos that were hypoxic from day 0 to 10 weighed less than control embryos at day 15 (P <.05), but not at hatch. At hatch, neither hypoxia nor malnutrition decreased crown-rump length. Brain and heart weights were increased in both malnourished groups, but not chicks that were hypoxic from day 0. CONCLUSION: Chick embryos exposed to malnutrition show asymmetric growth restriction with relative sparing of the brain and heart. Early growth restriction that was induced by hypoxia from the beginning of incubation is reversed by the restoration of normoxia at mid incubation. PMID- 11854648 TI - Elevated amniotic fluid C-reactive protein at the time of genetic amniocentesis is a marker for preterm delivery. AB - OBJECTIVE: A pre-existing intrauterine inflammation in the first half of gestation has been proposed as a possible condition that leads to preterm delivery. Indeed, elevated levels of inflammatory mediators (eg, interleukin-6, tumor necrosis factor) in midtrimester amniotic fluid have been found in cases of preterm delivery and/or spontaneous abortion. The objective of this study was to investigate whether the amniotic fluid C-reactive protein level at the time of genetic amniocentesis is a marker for spontaneous preterm delivery before 34 and 37 weeks of gestation. STUDY DESIGN: Women who underwent genetic amniocentesis between 15 and 18 weeks of gestation with (1) singleton gestation, (2) uneventful pregnancy course before the amniocentesis, and (3) absence of fetal abnormalities were included in the study. Patients with abnormal karyotype were excluded. C reactive protein concentration was measured in amniotic fluid and in maternal blood immediately after genetic amniocentesis. All patients were followed until delivery for the occurrence of pregnancy complications. Nonparametric tests and receiver-operating characteristic curve analysis were used for statistical purposes. RESULTS: The prevalence of spontaneous preterm delivery before 34 and 37 weeks was 3.3% (10 of 306 pregnancies) and 8.5% (26 of 306 pregnancies), respectively. Women with preterm delivery at <37 weeks had a higher median (range) of amniotic fluid C-reactive protein concentration than those women who delivered at term (median, 113.3 ng/mL [range, 16-623 ng/mL] vs median, 57.8 ng/mL [range, 0-808.9 ng/mL]; P <.005). Women with preterm delivery at <34 weeks had a higher median (range) amniotic fluid C-reactive protein concentration than those women who delivered at term (median, 183.8 ng/mL [range, 46.5-447 ng/mL] vs median, 57.8 ng/mL [range, 0-808.9 ng/mL]; P <.005]. No correlation was found between amniotic fluid C-reactive protein and maternal blood C-reactive protein concentrations. No relationship was found between maternal blood C-reactive protein concentration and preterm delivery before either 34 or 37 weeks. Amniotic fluid C-reactive protein concentration of >110 ng/mL had a sensitivity of 80.8% and a specificity of 69.5% in the prediction of spontaneous preterm delivery at <34 weeks. CONCLUSION: This study supports the theory that a subclinical intrauterine/fetal inflammatory process early in gestation may be important for the occurrence of preterm delivery in the second half of gestation. PMID- 11854649 TI - Parity and pregnancy outcomes. AB - OBJECTIVE: The issue of parity and risk of pregnancy outcomes has been controversial for many decades. We conducted a study to evaluate the association between parity and pregnancy outcomes and to clarify the basis of the classification of risk based on parity. STUDY DESIGN: The study is a cross sectional analysis on a large routinely collected data set of singleton births from 1992 to 1997 in New South Wales, Australia. Pregnancy outcomes were compared among parity groups. RESULTS: There was a significant association between different parity levels and pregnancy outcomes in terms of obstetric complications, neonatal morbidity, and perinatal mortality, after adjustment for potential confounders, including age, maternal smoking status, and socioeconomic status. In terms of obstetric complications, neonatal morbidity, and perinatal mortality, subjects can be classified into 3 groups according to parity: nulliparity, low multiparity (parity 1, 2, and 3), and grand multipara (parity 4 to 8). Compared with low multiparity, mothers and babies of nulliparity and grand multipara are at higher risk. CONCLUSIONS: This study supported the definition of grand multiparity that should start from parity 4 or the 5th baby. Attention should be paid to subjects of nulliparity and grand multiparity for their different problems. PMID- 11854650 TI - The prediction and prevention of intrapartum fetal asphyxia in preterm pregnancies. AB - OBJECTIVE: This study was undertaken to examine the contribution of electronic fetal heart rate monitoring in the prediction and prevention of intrapartum fetal asphyxia in the preterm pregnancy. STUDY DESIGN: The outcome of 40 pregnancies with biochemically confirmed intrapartum fetal asphyxia (ie, an umbilical artery base deficit >12 mmol/L) was examined. This included 20 pregnancies that were delivered abdominally matched with 20 pregnancies that were delivered vaginally. Antepartum and intrapartum clinical risk factors and newborn complications were documented. The interpretation of the fetal heart rate record as determined by the responsible clinicians was recorded. Fetal asphyxia was classified as mild, moderate, or severe on the basis of an umbilical artery base deficit of >12 mmol/L and newborn encephalopathy and other organ system complications. RESULTS: Fetal asphyxia was classified as mild in 21 pregnancies and as moderate or severe in 19 pregnancies. The fetal heart rate record was predictive of fetal asphyxia in 27 pregnancies and nonpredictive in 11 pregnancies. The predictive fetal heart rate record was the primary indication for intervention in 21 of the 24 pregnancies that were delivered by cesarean delivery in the first stage or operative vaginal delivery in the second stage of labor. Newborn outcome may have been influenced by intervention and delivery because of a predictive fetal heart rate record in 10 pregnancies with mild fetal asphyxia and in 9 pregnancies with moderate or severe fetal asphyxia. CONCLUSION: Electronic fetal monitoring is a useful screening test for the prediction of intrapartum fetal asphyxia in the preterm pregnancy. A predictive fetal heart rate record was the principle indication that led to intervention and delivery. A prediction of fetal asphyxia that leads to intervention and delivery may prevent or modify moderate or severe newborn morbidity as the result of fetal asphyxia. PMID- 11854652 TI - Erythromycin use during pregnancy in relation to pyloric stenosis. AB - OBJECTIVE: Newborn infants treated with erythromycin may be at risk for developing pyloric stenosis. Because erythromycin is known to cross the placenta and is a recommended treatment for chlamydia and other infections in pregnancy, we explored whether erythromycin taken during pregnancy might similarly lead to an increase in risk of pyloric stenosis. STUDY DESIGN: We used data collected between 1976 and 1998 as part of an ongoing case-control surveillance program. Cases were 1044 infants with a diagnosis of pyloric stenosis. Two control groups were used: 1704 nonmalformed infants and 15,356 infants with a wide range of other malformations. Odds ratios and 95% CIs were calculated by using data from each control group. RESULTS: All odds ratio estimates are close to 1.0, all CIs include 1, and all upper 95% confidence bounds are less than 2.0. CONCLUSION: We found no evidence of an increased risk of pyloric stenosis among infants born to mothers exposed to erythromycin during pregnancy. PMID- 11854651 TI - Maternal serum activin A, inhibin A, and follistatin in pregnancies with appropriately grown and small-for-gestational-age fetuses classified by umbilical artery Doppler ultrasound. AB - OBJECTIVE: The purpose of this study was to examine the relationship of maternal serum activin A, inhibin A, and follistatin with fetal growth and placental function. STUDY DESIGN: Inhibin A, activin A, and follistatin were measured in maternal serum that was stored from normally grown (control subjects, n = 50) and small-for-gestational-age pregnancies (n = 49), prospectively classified as normal small-for-gestational-age pregnancy or fetal growth-restricted pregnancy with the use of umbilical artery Doppler ultrasound. RESULTS: Activin A and inhibin A were significantly increased in fetal growth-restricted pregnancies compared with control subjects (activin A: regression coefficient, 0.54, P <.001; inhibin A: regression coefficient, 0.47, P =.003). The activin:follistatin ratio was significantly higher in fetal growth-restricted pregnancies compared with control subjects (P <.001). There were no significant differences between analyte levels of normal small-for-gestational-age pregnancies and control subjects. CONCLUSION: Maternal serum activin A, inhibin A, and activin:follistatin ratio are raised in fetal growth-restricted pregnancies but not in normal small-for gestational age pregnancies. This provides further evidence of the difference between subgroups within small-for-gestational-age pregnancies and emphasizes the need to stratify for this in research. PMID- 11854653 TI - Placental basal plate formation is associated with fibrin deposition in decidual veins at sites of trophoblast cell invasion. AB - OBJECTIVE: We hypothesize that placenta growth and basal plate (Nitabuch's fibrinoid) formation involves fibrin deposition in decidual veins as trophoblast cells invade. STUDY DESIGN: Decidua from women who underwent first trimester elective pregnancy termination (n = 15 women), in situ pregnancy (n = 2 women), and ectopic pregnancy (n = 5 women) were immunostained with polyclonal antibodies to fibrin. Serial sections of decidua were studied with monoclonal antibodies to oncofetal fibronectin, cytokeratin, and Factor VIII-related antigen. RESULTS: In all intrauterine pregnancies, trophoblast cells were identified with clots in decidual veins. Fibrin and oncofetal fibronectin were present in veins at sites of trophoblast cell invasion and in the developing basal plate matrix where villi had implanted into veins. Fibrin deposits were not seen in decidual parietalis of ectopic or in situ pregnancies. CONCLUSION: We have identified fibrin in decidual veins that is associated with trophoblast cell invasion. We speculate that the formation of basal plate protein (Nitabuch's fibrinoid) involves these intravenous fibrin deposits. PMID- 11854654 TI - Investigation of intraplacental villous arteries by Doppler flow imaging in growth-restricted fetuses. AB - OBJECTIVE: The purposes of our study were to assess the ability of color and power Doppler sonography to depict the blood flow in the intraplacental villous arteries and to evaluate whether the blood flow of intraplacental villous arteries in a normal pregnancy is different from that in a pregnancy that is associated with intrauterine growth restriction. STUDY DESIGN: Eighty-five women with uncomplicated pregnancy and 16 women with intrauterine growth-restricted fetuses between 27 and 38 weeks of gestation were examined by color and power Doppler imaging. The blood flow of intraplacental villous arteries was analyzed comparatively. The pulsatility index and peak systolic velocity were measured. RESULTS: A unit of 1 intraplacental villous artery-1 and its branches were seen as 1 cotyledon by color and power Doppler imaging. The cotyledon was easily identified and counted. Each cotyledon contained only 1 intraplacental villous artery-1. This method can visualize the intraplacental villous artery-1 to intraplacental villous artery-4 in normal pregnancies. The terminal villous arteries beyond intraplacental villous artery-4 were not imaged. The number of detectable intraplacental villous artery-1 in 1 placenta in intrauterine growth restriction was significantly lower than that in normal pregnancy. The number of detectable branches in intrauterine growth restriction was also significantly lower than in normal pregnancy. No intraplacental villous artery-4 blood flow was found in women with intrauterine growth restriction. In examined arteries, pulsatility index decreased and peak systolic velocity increased significantly with advancing gestational age (P <.02). At any given gestational age, pulsatility index and peak systolic velocity in the peripheral arteries were significantly lower than those in the upstream arteries in normal pregnancy (P <.001). The pulsatility index value of each intraplacental villous artery was also lower than that of the umbilical artery in the women with intrauterine growth restriction (P <.05). There were no differences in pulsatility index in each artery between the groups, although there were a few high pulsatility index values in intrauterine growth restriction. CONCLUSION: Color Doppler and power flow sonography are valuable tools for the detection of the blood flow of intraplacental villous arteries. The decrease in the number of detectable intraplacental villous artery-1 and branches was associated with intrauterine growth retardation. PMID- 11854655 TI - Prolonged hypoxia upregulates vascular endothelial growth factor messenger RNA expression in ovine fetal membranes and placenta. AB - OBJECTIVE: In ovine fetuses, 4 days of hypoxia resulted in a large increase in urine flow, without the development of polyhydramnios, which suggests that intramembranous absorption of the amniotic fluid was enhanced. Because vascular endothelial growth factor is speculated to be a regulator of intramembranous absorption through increases of membrane vascularity and fluid transport, we hypothesized that hypoxia upregulated vascular endothelial growth factor gene expression in the fetal membranes. STUDY DESIGN: Five near-term ovine fetuses that were subjected to 4 days of hypoxia and 5 age-matched time controls were studied. On day 4, the amnion, chorion, and placenta were collected for cellular localization and quantification of vascular endothelial growth factor messenger RNA and for the determination of vascular endothelial growth factor molecular forms that were expressed. The data were analyzed statistically with the use of t tests and 2-factor analyses of variance. RESULTS: Vascular endothelial growth factor messenger RNA was expressed in the fetal membranes localized to the amniotic epithelium and chorionic cytotrophoblast, and to the villous cytotrophoblast of the placenta. In hypoxic fetuses, vascular endothelial growth factor messenger RNA levels in these cell layers were significantly increased compared with the controls. Five vascular endothelial growth factor molecular forms were identified with vascular endothelial growth factor(164) being the most abundant form expressed. The pattern of expression of the forms was not altered by hypoxia. CONCLUSION: In the near-term ovine fetus, hypoxia induced vascular endothelial growth factor messenger RNA expression in the amnion, chorion, and placenta. This was associated with an increase in intramembranous absorption of amniotic fluid. We speculate that the increased intramembranous absorption was mediated by a vascular endothelial growth factor-induced increase in the transport of amniotic fluid into the fetal membranes. PMID- 11854656 TI - Neonatal morbidity associated with uterine rupture: what are the risk factors? AB - OBJECTIVE: The purpose of this study was to evaluate factors associated with severe metabolic acidosis or death in a situation of uterine rupture. STUDY DESIGN: A retrospective study was performed with review of charts and electronic monitoring strips. RESULTS: Between November 1988 and November 2000, a total of 23 cases of complete uterine rupture were identified from among 2233 trials of labor after a previous low transverse cesarean delivery. Nine neonates (39.1%) had severe metabolic acidosis (pH < 7.0); among these, hypoxic-ischemic encephalopathy was diagnosed in 3 neonates and another neonate died. Placental or fetal extrusion or both were associated with severe metabolic acidosis (P <.001) but not with the other factors (birth weight, induction of labor, use of oxytocin, use of epidural, and cervix dilatation). Two newborns with severe acidosis had impaired motor development even with an intervention time less than 18 minutes from the onset of prolonged deceleration to delivery. CONCLUSION: When uterine rupture occurs, placental or fetal extrusion was the most important factor associated with severe metabolic acidosis. Prompt intervention did not always prevent severe metabolic acidosis and neonatal morbidity. PMID- 11854658 TI - Ultrasonographic measurement of the abdominal circumference in fetuses with congenital diaphragmatic hernia. AB - OBJECTIVE: To determine whether ultrasonographic measurements of the abdominal circumference are smaller in fetuses with congenital diaphragmatic hernia and whether this is reflected as an underestimation of the estimated fetal weight. STUDY DESIGN: A retrospective review of 225 abdominal circumference measurements made between 24 and 41 weeks of gestation in 85 fetuses with congenital diaphragmatic hernia was performed. The individual and mean abdominal circumference value at each week of gestation versus gestational age was plotted and compared with normative data. Comparisons between abdominal circumference measurements and hernia variables were made with the chi(2) test. The Pearson correlation was used to examine the accuracy of ultrasonographic determination of the estimated fetal weight. RESULTS: The mean measurements of abdominal circumference were not found to differ significantly from normative data until term, although fetuses with liver herniation were less likely to have measurements more than 2 standard deviations below the mean. Calculation of estimated fetal weight was similar in accuracy to that in normal fetuses. CONCLUSION: Small abdominal circumference measurements should not be expected in fetuses with congenital diaphragmatic hernia. Abnormalities of the abdominal circumference or an abdominal circumference-dependent estimated fetal weight should not be attributed to the anatomic defect without considering other etiologies. PMID- 11854657 TI - Red cell mass responses to daily erythropoietin and iron injections in the ovine fetus. AB - OBJECTIVE: Anemic ovine fetuses supplemented with intra-amniotic iron undergo rapid expansion of red cell mass. The present study tested the hypothesis that nonanemic fetuses that were supplemented with daily intra-amniotic iron plus intravascular injections of erythropoietin would experience accelerated erythrocyte production. STUDY DESIGN: Nine late gestation ovine fetuses received 100 to 120 units of erythropoietin intravascularly plus 10 mg of iron intra amniotically daily for 7 days (low erythropoietin dose group). Four additional fetuses received 1000 units of erythropoietin plus 10 mg iron daily for the same period (high erythropoietin dose group). Responses were compared with 9 nonsupplemented time-control fetuses. Statistical testing was by 3-factor repeated measures analysis of variance. RESULTS: Immediately after erythropoietin injection, plasma erythropoietin concentration was elevated approximately 25- and 250-fold in the low and high erythropoietin dose groups, respectively. Erythropoietin returned to basal levels by 24 hours after the injection. Plasma iron concentration increased in the low erythropoietin dose group but not in the control or high erythropoietin dose groups. Reticulocyte index increased in both erythropoietin supplemented groups but not in control fetuses. Hematocrit level increased above control by day 5 in the low erythropoietin dose group and by day 2 in the high erythropoietin dose group. Red cell mass increased significantly on supplement day 7 in the low erythropoietin dose group and on day 5 in the high erythropoietin dose group. Fetal blood gases and pH were unchanged with time in all 3 groups. CONCLUSION: Although daily combined erythropoietin and iron supplements in nonanemic ovine fetuses significantly increased circulating red cell mass in a dose-dependent manner, this increase was small relative to the rapid expansion of red cell mass previously observed after iron supplementation in fetuses with hemorrhage-induced anemia. We speculate that this difference in response may be due to a combination of rapid fetal clearance of erythropoietin plus a relative insensitivity to erythropoietin caused by the absence of other cytokines, which are elevated during fetal anemia. PMID- 11854659 TI - Hormone replacement therapy and the risk of death from breast cancer: a systematic review. AB - OBJECTIVE: Recent studies have shown an increased relative risk of breast cancer with hormone replacement therapy for > or = 5 years. However, tumors in hormone replacement therapy users are more likely to be localized and of a favorable histologic condition. Our objective was to determine the effects of hormone replacement therapy on the risk of death from breast cancer. STUDY DESIGN: We searched the literature to identify studies that addressed hormone replacement therapy and breast cancer death among healthy hormone replacement therapy users compared with nonusers. We included randomized controlled trials and cohort and case-control studies with adequate controls and retrievable risk estimates. RESULTS: Ten observational studies met the inclusion criteria; these studies varied widely with regard to population, choice of control group, ascertainment of exposure and outcome, and adjustment for potential confounders. CONCLUSION: Most studies showed a reduced risk of death from breast cancer in hormone replacement therapy users compared with nonusers. In a few studies, this risk reduction was attenuated after prolonged use or prolonged duration of follow-up. Limited information on recency of use, dose, or use of combined hormone replacement therapy was available. PMID- 11854660 TI - Implementation of group B Streptococcus prevention strategy. PMID- 11854662 TI - Who should perform genetic amniocentesis? PMID- 11854661 TI - Is helical computed tomography 100% sensitive to diagnose acute appendicitis during pregnancy? PMID- 11854663 TI - Lingual forced eruption orthodontic technique: clinical considerations for patient selection and clinical report. AB - Lingual forced eruption is a treatment modality that minimizes esthetic impairment during orthodontic treatment. Use of this technique can enhance the acceptance of orthodontic treatment by adult patients and minimize unnecessary compromises in the treatment planning of submerged roots. In this clinical report, a treatment modality for forced eruption therapy is described, clinical considerations for patient selection are identified, and the advantages and limits of the lingual orthodontic technique are discussed. PMID- 11854664 TI - Use of connective tissue grafts to enhance the esthetic outcome of implant treatment: a clinical report of 2 patients. AB - The reestablishment of normal tissue volume, contour, and architecture around dental implants may require the use of osseous or soft tissue augmentation techniques. This article describes 2 patient treatments that highlight the use of connective tissue grafts to enhance the esthetic outcome of dental implant treatment. PMID- 11854665 TI - Use of heat-pressed, leucite-reinforced ceramic on anterior and posterior onlays: a clinical report. AB - The introduction of new techniques and materials in all-ceramic systems, together with improvements in resin bonding agents, has enabled the use of resin cements to directly bond ceramic crowns, veneers, and inlays/onlays to conservative tooth preparations. Directly bonded ceramic onlays can eliminate the need for conventional means of retention and allow the restoration of lost tooth structure with minimal tooth preparation. This article presents the use and short-term clinical performance of directly bonded, heat-pressed, leucite-reinforced ceramic onlays in both anterior and posterior areas of the mouth. PMID- 11854666 TI - Implant-supported, long-span fixed partial denture for a scleroderma patient: a clinical report. AB - Scleroderma poses many difficulties and challenges for the treating dentist. Until now, dental implant treatment for patients with scleroderma was limited to either complete overdentures or short-span fixed partial dentures. This clinical report describes the use of modified techniques to fabricate a long-span fixed prosthesis delivered after serial extractions and implant placement. PMID- 11854667 TI - Histologic evaluation of a threaded hydroxyapatite-coated root-form implant located at a dehisced maxillary site and retrieved from a human subject: a clinical report. AB - This clinical report describes the microscopic analysis of a threaded hydroxyapatite-coated root-form implant retrieved from an 89-year-old subject after 10 months of service. The implant was never loaded and was removed because it was nonrestorable. Clinically, the buccal area of the implant was covered by soft tissue, whereas the palatal side was covered by bone. Light microscopic evaluation revealed tight contact between hydroxyapatite and bone with no sign of coating dissolution. Osteocytes were present, and Haversian canals were in close proximity to the implant surface. The buccal side of the implant demonstrated mild to moderate inflammatory infiltrate and signs of hydroxyapatite coating dissolution. These observations suggest that hydroxyapatite coatings can resist degradation in contact with bone but may be more prone to dissolution in contact with soft tissue. PMID- 11854668 TI - Use of an individual traction prosthesis and distraction osteogenesis to reposition osseointegrated implants in a juvenile with ectodermal dysplasia: a clinical report. AB - This article describes the use of distraction osteogenesis to reposition osseointegrated maxillary implants in an adolescent girl with ectodermal dysplasia and oligodontia. The distraction of 2 osteotomized segments was controlled by a prosthesis fabricated specifically for this purpose. PMID- 11854669 TI - Relationship between tooth shade value and skin color: an observational study. AB - STATEMENT OF PROBLEM: There is limited scientific information on the relationship between tooth shade and skin color. This lack of knowledge may impact the ability of the prosthodontist to select artificial teeth that complement the facial complexion of the patient. PURPOSE: This observational study explored the possibility of a relationship between tooth shade and skin color. MATERIALS AND METHODS: A total of 119 individuals aged 18 to 80 years participated in this observational study. Two investigators calibrated for examining tooth shade performed all examinations. A Vita-Lumin shade guide was used to examine either the maxillary right or left central incisor, one of which had to be restoration- and caries-free for study inclusion. Tooth shades were divided into 4 categories according to value, and skin tones were divided into 4 categories (fair, fair/medium, medium, and dark) with the use of L'Oreal True Illusion compact makeup shades as a guide. Categorical modeling with chi-square analysis and Fisher's exact tests was used to analyze the data (alpha=.05). RESULTS: No interaction among age, skin color, and tooth shade or gender, skin color, and tooth shade was found; however, age was associated with tooth shade (P<.001). Older persons were more likely to have teeth with lower values (darker). Among those aged 60 and older, 85% had teeth in the medium and low range values compared with 17% of those younger than 31. Significant tooth shade differences were discovered among patients with different skin colors (P<.05). Among those with low values (darker teeth), 50% were of fair complexion and 17% were of dark complexion (P<.001). CONCLUSION: Within the limitations of this study, tooth shade value and skin color were inversely related. Older adults were more likely to have darker teeth (lower value). PMID- 11854670 TI - The effect of two different coatings on the surface integrity and softness of a tissue conditioner. AB - STATEMENT OF PROBLEM: When they are used to treat inflamed, irritated, or distorted tissues or in implant therapy, tissue conditioners are required to function over relatively long time periods. Most presently available materials have a short life span (3 to 4 days). PURPOSE: This study evaluated the effects of 2 different coatings on the surface integrity and softness of a tissue conditioner over a 4-week period. MATERIAL AND METHODS: Ten subjects with a complete maxillary denture and a functioning mandibular dentition were included in the study. Each maxillary denture had 3 troughs filled with a tissue conditioner (Coe Comfort). One trough was coated with Monopoly and one with Permaseal; the third trough was left uncoated (control). The specimens were tested at insertion and at 1 week, 2 weeks, and 4 weeks post-insertion for softness with a durometer and for surface integrity with a scanning electron microscope. Statistical differences were determined with analysis of variance (P<.05). RESULTS: At the end of 4 weeks, the mean and standard deviation of softness on the ASTM scale was 85.1 (4.2) for the control, 81.6 (3.9) for Monopoly, and 77.9 (5.0) for Permaseal. The difference between the control and Permaseal was significant (P<.0042). Qualitatively, SEM analysis indicated that the control deteriorated by the end of the first week, whereas both Monopoly and Permaseal coatings remained intact for up to 2 weeks. All groups showed some conditioner deterioration by 4 weeks. The tissues under the denture-bearing areas remained healthy during the study period. CONCLUSION: Within the limitations of this study, the application of Permaseal or Monopoly coatings significantly reduced the loss of tissue conditioner softness. Permaseal-coated conditioner remained the softest over the length of the study. PMID- 11854671 TI - High-speed cavity preparation techniques with different water flows. AB - STATEMENT OF PROBLEM: Cavity and tooth preparations generate heat because the use of rotary cutting instruments on dental tissues creates friction. Dental pulps cannot survive temperature increases greater than 5.5 degrees C. PURPOSE: This study evaluated the efficiency of 3 different water flows for 2 different tooth preparation techniques to determine which are safe for use. MATERIAL AND METHODS: Thermocouples were placed in the pulpal chambers of 30 bovine teeth, and 1 of 2 tooth preparation techniques was used: a low-load intermittent tooth preparation technique or a high-load tooth preparation technique without intervals. Water flows of 0, 30, and 45 mL/min were associated with each technique, for a total of 6 different groups. The results were analyzed with a 2-factor analysis of variance (P<.05). RESULTS: Temperature increases with the high-load technique were 16.40 degrees C without cooling (group I), 11.68 degrees C with 30 mL/min air-water spray cooling (group III), and 9.96 degrees C with 45 mL/min cooling (group V). With the low-load tooth preparation technique, a 9.54 degrees C increase resulted with no cooling (group II), a 1.56 degrees C increase with 30 mL/min air-water spray cooling (group IV), and a 0.04 degrees C decrease with 45 mL/min cooling (group VI). The low-load technique was associated with more ideal temperature changes. CONCLUSION: The results of this study confirm the necessity of using a low-load technique and water coolants during cavity and tooth preparation procedures. PMID- 11854672 TI - Compositional stability and marginal accuracy of complete cast crowns made with as-received and recast type III gold alloy. AB - STATEMENT OF PROBLEM: The effect of combining new and recast metal on the compositional stability of the alloy used and accuracy of the final restoration is not known. PURPOSE: This study characterized the elemental compositional stability of as-received and recast type III gold alloy. The effect of combining these alloys on the marginal accuracy of complete cast crowns also was evaluated. MATERIAL AND METHODS: Sixty standardized type III gold alloy complete crowns were made on epoxy resin dies duplicated from a metal master die with conventional laboratory techniques. Three casting protocols (n = 20 each) were established in relation to the proportion of as-received and recast gold alloys: Group A (100% as-received metal), Group B (50% wt new metal, 50% wt once recast metal), and Group C (100% once recast metal). An elemental analysis was performed at 3 sites along the cast crown (cervical, middle, and occlusal) with x-ray energy dispersive spectroscopy. Marginal accuracy was measured with a microscope before and after cementation. Results were subjected to 1-way analysis of variance followed by the Ryan-Einot-Gabriel-Welsch multiple-range test at a 1% level of significance. RESULTS: Elemental composition was significantly different among the casting groups (P<.001). The mean weight percentage values were 72.4% to 75.7% Au, 4.5% to 7.0% Pd, 10.7% to 11.1% Ag, 7.8% to 8.4% Cu, and 1.0% to 1.4% Zn. Statistically but not clinically significant differences also were found for marginal accuracy. The marginal discrepancy was less than 25 microm for all casting conditions, with the lowest value recorded for Group A (7 microm), the highest for Group C (12 microm), and an intermediate value for Group B (9 microm) specimens. CONCLUSION: Recasting type III gold alloys may adversely affect the marginal accuracy of complete cast crowns. In this study, however, such effects were not clinically significant. PMID- 11854673 TI - Effect of posterior crown margin placement on gingival health. AB - STATEMENT OF PROBLEM: The clinical impact of posterior crown margin placement on gingival health has not been thoroughly quantified. PURPOSE: This study evaluated the effect of posterior crown margin placement with multivariate analysis. MATERIAL AND METHODS: Ten general dentists reviewed 240 patients with 480 metal ceramic crowns in a prospective clinical trial. The alloy was randomly selected from 2 high gold, 1 low gold, and 1 palladium alloy. Variables were the alloy used, oral hygiene index score before treatment, location of crown margins at baseline, and plaque index and sulcus bleeding index scores recorded for restored and control teeth after 1 year. The effect of crown margin placement on sulcular bleeding and plaque accumulation was analyzed with regression models (P<.05). RESULTS: The probability of plaque at 1 year increased with increasing oral hygiene index score before treatment. The lingual surfaces demonstrated the highest probability of plaque. The risk of bleeding at intrasulcular posterior crown margins was approximately twice that at supragingival margins. Poor oral hygiene before treatment and plaque also were associated with sulcular bleeding. Facial sites exhibited a lower probability of sulcular bleeding than lingual surfaces. Type of alloy did not influence sulcular bleeding. CONCLUSION: In this study, placement of crown margins was one of several parameters that affected gingival health. PMID- 11854674 TI - Peri-implant tissue response of immediately loaded, threaded, HA-coated implants: 1-year results. AB - STATEMENT OF PROBLEM: Although high success rates have been reported with immediately loaded implants, the peri-implant tissue response has not been well documented. PURPOSE: This study evaluated implant success and peri-implant tissue response of immediately loaded, threaded, hydroxyapatite (HA)-coated root-form implants supporting mandibular bar overdentures with opposing conventional maxillary complete dentures in humans. MATERIAL AND METHODS: Five patients (3 men, 2 women; mean age 61 years) each received 4 HA-coated endosseous root-form implants in the interforaminal region in the mandible. The implants were rigidly splinted with a metal framework within 24 hours. The final EDS clip prosthesis was placed 1 to 2 weeks thereafter. The implants and peri-implant tissues were evaluated clinically and radiographically 0, 1, 3, 6, and 12 months after prosthesis placement. Data were analyzed with a repeated measures 1-way analysis of variance (P<.05). RESULTS: All implants were stable at the end of the observation period (mean Periotest value = -5.9 +/- 1.4). No peri-implant radiolucencies were noted, and no implants were lost. The mean marginal bone changes were -0.42 plus minus 0.34, -0.84 +/- 0.55, -1.14 +/- 0.80, and -1.16 +/- 0.89 mm at the 1-, 3-, 6-, and 12-month follow-ups, respectively (P<.001). Significant declines in the rates of marginal bone changes at each time interval were noted (P<.001). In addition, there were significant decreases in probing depth (P<.001) and plaque index (P<.001) but no significant difference in the frequency of bleeding upon probing (P=.64). CONCLUSION: Within the limitations of this study, the peri-implant tissue response of immediately loaded, HA-coated implants was favorable and comparable to that of conventional, delayed-loaded implants after 1 year. PMID- 11854675 TI - Changes in dentin after insertion of self-threading titanium pins with 3 methods: a scanning electron microscope pilot study. AB - STATEMENT OF PROBLEM: Inserting a self-threading pin can cause dentinal cracks, but it is not known whether dentin bonding agents can penetrate these cracks. PURPOSE: Part I of this in vitro pilot study was conducted to document the presence of dentinal cracks after the placement of self-threading pins with 3 methods. Part II was conducted to observe changes in dentin when a dentin bonding agent was applied before insertion of the self-threading pins. MATERIAL AND METHODS: The crowns of 14 noncarious third molars were sectioned horizontally 2 mm above the cemento-enamel junction, and the occlusal portions were discarded. The teeth were put into a nontransparent bag and divided randomly into 2 groups. Group 1 consisted of 12 teeth. In each tooth, 4 pinholes were prepared with a handpiece at normal rotation speed (30,000 rpm). Self-threading pins were placed into 3 of the pinholes in each tooth: 1 manually, 1 by handpiece at 7000 rpm, and 1 by handpiece at 30,000 rpm. The fourth pinhole was left empty and served as the control. All group 1 teeth were sectioned vertically through the pins, dental hard tissue, and control pin holes. The examination surface of each specimen in group 1 was polished, and the smear layer was removed with Calcinase and NaOCl solutions. After dehydration in ascending grades of alcohol, specimens were coated with a 10- to 15-nm-thick layer of gold and examined with a scanning electron microscope. In the remaining 2 teeth (group 2), a dentin bonding agent was introduced into the pinholes prior to pin placement. Two pins were placed manually and 2 by handpiece at 30,000 rpm. After pin placement, sectioning, cleaning, and dehydration, the specimens were examined with a scanning electron microscope, and x-ray mapping was performed to determine the presence of titanium, silicon, and calcium. RESULTS: Craze lines in dentin were associated with 54.5% of pins placed manually and 54.5% of pins placed with a handpiece at reduced speed (7000 rpm). Dentinal cracks were associated with 50% of pins placed with a handpiece at standard speed and with 16.7% of the control pinholes. X-ray mapping analysis revealed the presence of the dentin bonding agent between the pin and dentin wall. The dentin bonding agent was not found in the dentinal cracks except at the crack orifice. SUMMARY: Within the limitations of this pilot study, the method of pin insertion had no direct bearing on the presence of dentinal cracks. The dentin bonding agent tested did not fill the entire space of dentinal cracks but did occlude their orifices and fill the spaces between pin surface and dentin walls in the pin preparation. PMID- 11854676 TI - Clinical evaluation of all-ceramic crowns. AB - STATEMENT OF PROBLEM: There are few in vivo studies on the clinical performance of all-ceramic crowns. PURPOSE: The aim of this study was to evaluate the clinical performance of IPS Empress crowns. MATERIAL AND METHODS: Thirty-seven all-ceramic IPS Empress crowns were placed in 20 patients. Twenty crowns were luted with Variolink II low-viscosity resin cement in combination with Syntac Classic dentin adhesive; the remaining 17 were luted with Variolink II in combination with Syntac Single Component. All procedural steps were performed by the same prosthodontist. Using the California Dental Association's (CDA) quality evaluation system, 2 calibrated evaluators examined the crowns for margin integrity, anatomic form, surface, and color for a period of 12 to 41 months, with a mean of 24.56 months after insertion. Kaplan-Meier statistical analysis was used to calculate the survival rate of the crowns. Values obtained for plaque and gingival conditions were compared to control teeth with use of the Wilcoxon signed ranks test (P<.05). RESULTS: Based on the CDA criteria, 94.6% of the crowns were rated satisfactory. Fracture was registered in only 1 crown. One endodontically treated tooth failed due to the dislodgement of the prefabricated dowel. No significant difference was observed in the gingival health status of crowns that had margins placed above or at the level of the gingival margin. However, in crowns that had subgingival margin finish lines, the percentage of bleeding on probing was significantly higher than that of the contralateral control teeth. CONCLUSION: In this in vivo study, IPS Empress crowns luted with both dentin bonding agents functioned satisfactorily with a relatively low fracture rate over a mean evaluation period of 24.56 months. PMID- 11854677 TI - No association between incisal tooth wear and temporomandibular disorders. AB - STATEMENT OF PROBLEM: Incisal tooth wear may be a sign of long-term bruxing behavior. Bruxism is purported to be a risk factor for temporomandibular disorders (TMD). PURPOSE: The purpose of this clinic-based case-control study was to investigate whether the wear of anterior teeth is associated with TMD. MATERIAL AND METHODS: Two hundred eight TMD patients and 172 control subjects were selected for participation. After exclusion of subjects with more than 1 missing premolar or molar zone in opposite arches, as well as subjects with missing or severely restored anterior teeth, 154 TMD patients and 120 control subjects were included in the study (age 31.2 +/- 13.4 years; range 13 to 76 years). Anterior tooth wear was assessed on casts with a 0 to 5 scale. A multiple logistic regression analysis, controlling for the effects of age and gender, was performed to investigate the relationship between tooth wear and TMD. RESULTS: An odds ratio of 0.76 (95% confidence interval: 0.51 to 1.15) indicated that after adjusting for gender and age, the odds in favor of TMD decreased an estimated 24% for each additional unit of the mean tooth wear score. This result was not significant (P=.20). CONCLUSION: Within the limitations of this study, incisal tooth wear (assessed on dental casts) was not significantly associated with TMD when the influence of age and gender was controlled. Based on these findings, a clinically relevant risk for TMD from incisal tooth wear can be excluded. Hence, the presented evidence does not support the idea that treatment of incisal tooth wear is indicated to prevent TMD. PMID- 11854678 TI - Laser digitization of casts to determine the effect of tray selection and cast formation technique on accuracy. AB - STATEMENT OF PROBLEM; Several studies have attempted to determine the ideal combination of dental materials and laboratory techniques to produce the most accurate dental cast. Most have made use of 2-dimensional manual measuring devices, which neglect to account for the dimensional changes that exist along a 3-dimensional surface. PURPOSE: The purpose of this study was to determine the effect of impression tray selection and cast formation techniques on the dimensional change of a dental cast with the use of new, 3-dimensional optical digitizing technology. MATERIAL AND METHODS: Multiple impressions of a machined steel die that resembled a dental arch were made with custom and stock impression trays and vinyl polysiloxane impression material. The impressions were poured in type V artificial dental stone and allowed to set with the tray inverted or noninverted. The steel master die and stone casts were digitized with the Steinbichler Comet 100 Optical Digitizer, which was developed at the Minnesota Dental Research Center for Biomaterials and Biomechanics. Three-dimensional images of the stone casts were aligned to the 3-dimensional image of the master die and analyzed with AnSur-NT software. Multiple measurements of the master die and stone casts were analyzed to determine the accuracy of the 3-dimensional technology and of the impression and cast fabrication techniques. Planar distances between the center of each crown preparation were measured, as were crown heights. Data were analyzed with analysis of variance (P<.05), and root mean square error values were determined. RESULTS: Casts were compared with a total of 45 significance tests, of which only 4 yielded P<.05. There was no pattern to these results, which suggests that they were false-positive findings. CONCLUSION: Results obtained with the use of new optical digitizing technology indicated that neither impression tray type nor cast formation technique affected the accuracy of final casts. PMID- 11854679 TI - Three-dimensional assessment of the reliability of a postural face-bow transfer. AB - STATEMENT OF PROBLEM: Incorrect 3-dimensional orientation of a dental cast may compromise the functional and esthetic result of prosthetic restorations. PURPOSE: This study assessed the reliability of a new method to transfer the 3 dimensional orientation of the occlusal plane with a postural face-bow. MATERIAL AND METHODS: The 3-dimensional position of the occlusal plane in 20 subjects (age 20 to 32 years) with a complete dentition in both arches was assessed with a postural face-bow. An irreversible hydrocolloid impression of the maxillary arch was made for each subject and poured in dental stone. The maxillary arch was mounted in an articulator with use of a postural face-bow. The 3-dimensional position of the occlusal plane was then measured and compared to the values obtained with the use of a previously certified, computerized, noninvasive instrument. This instrument digitizes the coordinates of dental and facial landmarks and then calculates the spatial position of the occlusal plane. For each subject, the direct assessment and the face-bow measurement were compared by calculating the absolute difference of the following: the inclination of the occlusal plane relative to the true vertical, frontal plane projection (angle alpha) and sagittal plane projection (angle beta); intercondylar distance; distance of the center of gravity of the anterior part of the maxillary arch to the midpoint of the intercondylar axis; and the maxillary right canine to right condylion distance. Descriptive statistics of the differences were calculated. RESULTS: The postural face-bow appeared reliable and compared well to the computerized assessment, with mean differences ranging from 2.5 degrees to 3 degrees. CONCLUSION: In the population tested, a postural face-bow reliably reproduced the spatial orientation of the occlusal plane relative to the true horizontal plane. This position was transferred to an articulator with limited errors. PMID- 11854680 TI - 4-META use in dentistry: a literature review. AB - 4-META (4-methacryloyloxyethy trimellitate anhydride) was invented and patented in Japan in January 1979 and patented in the United States in April 1979. For the first 8 years, 4-META dental products (manufactured by Sun Medical Co, Kyoto, Japan) were not readily available in North America. Most of the early research therefore was done in Japan and reported in Japanese. A second wave of products (manufactured by Parkell, Farmingdale, N.Y.) became available in the United States in 1987, which precipitated research in English. This article is a chronological review of the literature published between 1978 and 1998 on 4-META dental adhesive materials. Thirteen commercial products are reviewed in terms of their physical strength; research on the mechanisms of 4-META action and the biocompatibility of 4 META was excluded. PMID- 11854681 TI - Flexural strength of provisional crown and fixed partial denture resins. AB - STATEMENT OF PROBLEM: Provisional prostheses are subject to flexure under function. Selection of the appropriate material for their fabrication is difficult given the limited evidence-based information on the flexural strength of provisional resins. PURPOSE: This study compared the flexural strength of 5 methacrylate-based resins and 8 bis-acryl resins used to fabricate provisional crowns and fixed partial dentures. MATERIAL AND METHODS: Bar-type specimens were fabricated according to American National Standards Institute/American Dental Association specification 27. After being immersed in artificial saliva at 37 degrees C for 10 days, the specimens were fractured under 3-point loading in a universal testing machine at a crosshead speed of 0.75 mm/min. Maximal loads to fracture in Newtons were recorded. Mean flexural strengths were calculated in MPa (n = 10 per group). Comparisons were made with analysis of variance and Duncan's multiple range test (P<.05). RESULTS: Mean flexural strengths ranged from 56.2 to 123.6 MPa. There were 4 statistically similar groups. The group with the highest strengths consisted of 4 bis-acryl materials (Provipont, Integrity, Protemp 3 Garant, and Luxatemp). CONCLUSION: Within the limitations of this study, flexural strengths were material- rather than category-specific. Some, but not all, bis acryl resins demonstrated significantly superior flexural strength over traditional methacrylate resins. PMID- 11854682 TI - Use of transitional implants to support a surgical guide: enhancing the accuracy of implant placement. AB - Surgical guides for edentulous patients often lack the stability desired for accurate implant placement. The difficulty is emphasized in the edentulous mandible when implant-supported metal-ceramic restorations, which require precise placement of implants, are planned. A technique is presented for the fabrication of a surgical and radiographic template, supported by transitional implants, that guides the placement of conventional implants. This template can enhance placement accuracy in an efficient way to achieve predictable, esthetic results. PMID- 11854683 TI - A surgical guide for accurate mesiodistal paralleling of implants in the posterior edentulous mandible. AB - Malaligned implants often complicate the clinical and laboratory procedures employed for the fabrication of superstructures. Due to improper load distribution, an overall increase in stress concentrations on supporting implants may occur; in the long run, this may compromise the maintenance of the bone implant interface. The utilization of surgical stents is imperative for optimum implant installation. This article describes a modified surgical stent that serves as a guide to proper mesiodistal paralleling of dental implants. PMID- 11854684 TI - Modification of the Barton button for tracheoesophageal speech: an innovative maxillofacial prosthetic technique. AB - Laryngectomized, tracheoesophageal speakers who achieve hands-free speech wear a special breathing valve to automatically shunt air into the pharyngoesophagus for speech production. The standard method of attachment is adherence to the peristomal skin with glues and adhesives. The Barton button is a simpler, more efficient alternative that provides intraluminal attachment within the trachea. Although the device is appealing, constraints in standard design and dimensions have limited its success. This article presents a technique for modification of the Barton button to increase its successful use by tracheoesophageal speakers. PMID- 11854685 TI - A modified occlusal plane device. PMID- 11854686 TI - Technique for removing a cemented superstructure from an implant abutment. PMID- 11854687 TI - Skills assessment of surgeons. PMID- 11854688 TI - Role of activation peptides from pancreatic proenzymes in the diagnosis and prognosis of acute pancreatitis. PMID- 11854689 TI - Myofibroblasts are responsible for the desmoplastic reaction surrounding human pancreatic carcinomas. AB - BACKGROUND: The cell type responsible for the desmoplastic reaction surrounding human pancreatic carcinoma is unknown. Hepatic stellate cells, which activate to a myofibroblast-like form, are responsible for collagen deposition in cirrhosis and around hepatocellular carcinomas. Recently, pancreatic stellate cells have been described and implicated in the fibrosis of chronic pancreatitis. We sought to determine whether these cells are responsible for the scirrhous reaction surrounding pancreatic adenocarcinomas. METHODS: Archival formalin-fixed, paraffin-embedded pancreatic tissues from 10 patients undergoing pancreaticoduodenectomy for ductal adenocarcinoma and from 2 patients with pancreatic islet cell tumors were examined immunohistochemically for alpha-smooth muscle actin (alpha-SMA), smooth muscle myosin heavy chain (SMMHC), procollagen I, collagen IV, and endothelial cell markers, von Willebrand factor and cluster of differentiation 31. RESULTS: In non-neoplastic areas, staining for alpha-SMA and SMMHC was confined to interlobular septal regions. In contrast, the desmoplastic reaction surrounding all 10 pancreatic adenocarcinoma specimens displayed intense interstitial staining for alpha-SMA, SMMHC, and collagen IV but no staining for von Willebrand factor and cluster of differentiation 31. Procollagen I staining localized intracellularly to fibroblast-shaped cells within this alpha-SMA/SMMHC-positive scirrhous region. Islet cell tumors demonstrated an increase in alpha-SMA staining, although this was not as marked as in ductal adenocarcinomas. CONCLUSIONS: A massive increase in myofibroblast activity, compatible with the activation of stellate cells, is associated with the deposition of collagen types I and IV in the desmoplastic reaction around pancreatic adenocarcinomas. PMID- 11854690 TI - Paucity of dendritic cells in pancreatic cancer. AB - BACKGROUND: The number of dendritic cells (DC) in the local tumor environment correlates with patient survival in numerous tumors. The relationship of DC infiltration in the tumor microenvironment and prognosis was examined in patients with pancreatic adenocarcinoma. METHODS: Forty-seven pancreatectomy specimens with a diagnosis of pancreatic adenocarcinoma were identified retrospectively and analyzed with the dendritic cell markers S-100 and CD1a. Patient survival was correlated with these markers and with p53, CD3, CD20, CD68, Ki-67. RESULTS: Significant numbers (>3 per high-powered field) of tumor-associated S100(+) or CD1a(+) cells were found in only 2/47 patients (4%). When present, dendritic cells were located outside the margin of the tumor. CD3, CD68, and CD20 positive cells were rare or absent in 96%, 92%, and 93% of the specimens. A correlation with survival and numbers of immune cells could not be made secondary to their rarity. The median survival was 18.9 months. No other indices measured correlated with survival. CONCLUSIONS: In patients with pancreatic adenocarcinoma, there is a paucity of immune cells within the tumor. PMID- 11854691 TI - Biliary sphincter motility after neural isolation of the pancreatoduodenal region in conscious dogs. AB - BACKGROUND: Several neural and hormonal factors are known to affect the motility of the sphincter of Oddi. However, the precise mechanisms of the control of sphincter motility have not been completely explored. We investigated the relationship of canine biliary sphincter motility when it is extrinsically denervated by neural isolation of the pancreatoduodenal region. METHODS: Interdigestive and postprandial sphincter motility in a denervated pancreatoduodenal segment and effects of cholecystokinin-octapeptide were studied in 7 conscious dogs. Data were compared with those of 7 neurally intact control dogs. RESULTS: After extrinsic denervation of the pancreatoduodenal region, sphincter motility exerted a cyclic change in concert with the duodenal myoelectric cycles; this change involved short cyclic bursts of motor activity, which gradually increased in intensity. The increase in the cyclic bursts of motor activity was also cyclic and associated with an increase in the plasma motilin concentration. Neural isolation of the pancreatoduodenal region increased sphincter basal pressure and motility index (integral per minute). In the denervated biliary sphincter, the feeding pattern and temporary inhibitory effect of feeding, as seen in controls, were absent, which suggests the role of extrinsic nerves in delivering bile into the duodenum after feeding. In the denervated dogs, cholecystokinin-octapeptide caused excitation of the sphincter activity, instead of relaxation observed in controls. CONCLUSIONS: Extrinsic innervation to the pancreatoduodenal region has an inhibitory effect on biliary sphincter motility. Abnormalities in extrinsic innervation to the biliary sphincter might increase the resistance of the sphincter to the bile flow and induce bile stagnation. PMID- 11854692 TI - Prospective evaluation of delayed technetium-99m sestamibi SPECT scintigraphy for preoperative localization of primary hyperparathyroidism. AB - BACKGROUND: Delayed technetium-99m sestamibi single photon emission computed tomography (SPECT) scans were prospectively analyzed in a large series of patients with primary hyperparathyroidism. METHODS: Three hundred thirty-eight patients underwent sestamibi-SPECT and were explored. Prospective data included preoperative demographics, clinical, sestamibi, and operative findings, laboratory values, and pathologic and follow-up laboratory results from all patients. RESULTS: Between 1994 and 2000, 287 unexplored patients (85%) and 51 re explored patients (15%) participated. The abnormal parathyroid glands excised from 336 of 338 patients included 299 single adenomas (88%) and 23 double adenomas (7%), and 14 patients had multigland hyperplasia (4%). Sestamibi SPECT correctly lateralized 349 of 400 abnormal parathyroid glands, with an overall sensitivity of 87%, an accuracy of 94%, and a positive predictive value of 86%. Precise localization occurred in 82% of the abnormal parathyroid glands. Sestamibi sensitivity was similar in unexplored (87%) and reoperative (92%) cases; two hundred eighty-six of 299 (96%) solitary adenomas, 38 of 46 (83%) double adenomas, but only 25 of 55 (45%) hyperplastic glands were identified. The mean weight of the true-positive glands (1252 +/- 1980 mg) was greater than that of the false-negative glands (297 +/- 286 mg) (P <.005). Three patients had persistent primary hyperparathyroidism, in spite of the excision of sestamibi identified lesions in 2 cases. Follow-up indicated curative resection in 99% of the unexplored cases and 94% of the remedial cases. CONCLUSIONS: Sestamibi SPECT is highly accurate for the localization of parathyroid adenomas in unexplored and re-explored cases, where it is often the only imaging required. Its sensitivity is limited in multiglandular disease. PMID- 11854693 TI - Endoscopic stapler-assisted diverticuloesophagostomy for Zenker's diverticulum: patient satisfaction and subjective relief of symptoms. AB - BACKGROUND: We assessed the results of endoscopic stapler-assisted diverticuloesophagostomy for Zenker's diverticulum, with emphasis on patient satisfaction and subjective relief of symptoms. METHODS: A retrospective cohort analysis was performed on all patients operated on from 1997 to 2000. Patients' charts were reviewed with respect to intraoperative and postoperative complications. The severity of preoperative and postoperative symptoms by patient self-assessment was compared by means of a 4-point scale (0-3) on a standardized questionnaire, as well as assessment of patient satisfaction (poor, fair, moderate, high). RESULTS: Thirty patients (22 men, 8 women) with a mean age of 72 years (range, 42-97 years) were part of the study. The mean follow-up was 13.2 months (range, 2-39 months). No major intraoperative or postoperative complications were observed. The most frequent preoperative symptoms were regurgitation of undigested food and dysphagia for solid consistencies (87%). Both of these preoperative symptoms were improved from a mean preoperative score of 2.2 and 2.7 to 0.1 and 0.4, respectively (P <.0001). Postprandial coughing (32%), halitosis (21%), and globus sensation (32%) were less common but also significantly improved. All but 1 patient (96%) were highly satisfied with the procedure. CONCLUSIONS: Endoscopic stapler-assisted treatment of Zenker's diverticulum offers a very low complication rate, a very high degree of symptom relief, and patient satisfaction. PMID- 11854694 TI - Delayed liver regeneration with negative regulation of hepatocyte growth factor and positive regulation of transforming growth factor-beta1 mRNA after portal branch ligation in biliary obstructed rats. AB - BACKGROUND: The influence of obstructive jaundice on liver regeneration is still controversial. The aim of this study was to investigate liver regeneration after portal branch ligation (PBL) in the jaundiced rat, focusing on hepatocyte growth factor (HGF) and transforming growth factor-beta1 (TGF-beta 1). METHODS: Male Wistar rats underwent PBL or a sham operation 7 days after a common bile duct ligation. Liver wet weight, proliferating cell nuclear antigen labeling, HGF and TGF-beta 1 mRNA expression, and immunohistochemical staining with alpha-smooth muscle actin antibody were studied. RESULTS: The rate of liver regeneration in jaundiced liver was decreased as compared to a non-jaundiced liver. DNA synthesis in the jaundiced non-ligated lobe was significantly lower than in the non jaundiced liver as was the peak level of HGF mRNA expression after PBL. In contrast, the level of TGF-beta 1 mRNA expression was higher in the jaundiced liver, and alpha-smooth muscle actin staining showed that hepatic stellate cells were gradually activated into myofibroblast-like cells. CONCLUSIONS: Obstructive jaundice decreased the expression of HGF mRNA and increased the expression of TGF beta 1 mRNA, resulting in delayed liver regeneration after PBL. We suggest that hepatic stellate cells activated in obstructive jaundice may affect the expression of these growth factors. PMID- 11854695 TI - Mitochondrial function during ischemic preconditioning. AB - Background. Ischemic preconditioning (IPC) protects the myocardium from ischemia reperfusion injury. The effect of IPC on the mitochondria is not well known. However, one of the mechanisms postulated in IPC (the opening of the mitochondrial K(ATP) channels) is likely to result in changes in mitochondrial function. Therefore, the purpose of this study was to determine the effect of IPC on mitochondrial function during ischemia reperfusion. Methods. Isolated rat hearts (n = 6/group) were subjected to (1) 30 minutes of equilibration, 25 minutes of ischemia, and 30 minutes of reperfusion (RP) (control group) or (2) 10 minutes of equilibration, two-5 minute episodes of IPC (each followed by 5 minutes of re-equilibration), 25 minutes of ischemia, and 30 minutes of RP (IPC group). Left ventricular rate pressure product (RPP) was measured. At end equilibration (end-EQ) and at end-reperfusion (end-RP) mitochondria were isolated. Mitochondrial respiratory function (state 2, 3, and 4), respiratory control index (RCI), rate of oxidative phosphorylation (ADP/Delta t), and ADP:O ratio were measured by polarography with the use of NADH- or FADH-dependent substrates. Results. IPC improved recovery of RPP at end-RP (72% +/- 5% in IPC vs 30% +/- 4% in control, P <.05). Ischemia reperfusion (IR) decreased state 3, ADP/Delta t, and RCI in both groups compared with end-EQ. IPC improved state 3 (47 +/- 3 in IPC vs 37 +/- 2 ng-atoms O/min/mg protein in control), ADP/Delta t (17 +/- 1 in IPC vs 13 +/- 1 nmol/s/mg protein in control), and RCI (3.7 +/- 0.1 in IPC vs 2.1 +/- 0.2 in control) at end-RP compared with control with the use of NADH-dependent substrate (P <.05 vs control). IPC also improved state 3 (85 +/- 6 in IPC vs 71 +/- 4 ng-atoms O/min/mg protein in control), ADP/Delta t (18 +/- 2 in IPC vs 12 +/- 1 nmol/s/mg protein in control), RCI (2 +/- 0.1 in IPC vs 1.5 +/ 0.1 in control), and ADP:O ratios (1.4 +/- 0.04 in IPC vs 1.7 +/- 0.09 in control) at end-RP compared with control with the use of FADH-dependent substrate (P <.05 vs control). Conclusions. The cardioprotective effects of IPC can be attributed at least in part to the preservation of mitochondrial function during reperfusion. PMID- 11854696 TI - Effectiveness of higher doses of botulinum toxin to induce healing in patients with chronic anal fissures. AB - BACKGROUND: Botulinum toxin induces healing in patients with idiopathic anal fissures. METHODS: One hundred-fifty patients with posterior anal fissures were treated with botulinum toxin injected in the internal anal sphincter on each side of the anterior midline. Subjects were randomized into 2 treatment groups based on the number of units of botulinum toxin injected. Patients in group I were treated with 20 units of botulinum toxin and, if the fissure persisted, were retreated with 30 units. Patients in group II were treated with 30 units and retreated with 50 units, if the fissure persisted. RESULTS: The 2 groups were comparable in age, gender distribution, duration of symptoms, resting pressure, and maximum voluntary pressure at anorectal manometry. One month after the injection, examinations revealed complete healing in 55 patients (73%) from group I and 65 patients (87%) from group II (P =.04). Five patients from group II reported a mild incontinence of flatus that lasted 2 weeks after the treatment and disappeared spontaneously. The values of the resting anal pressure (P=.3) and the maximum voluntary pressure (P =.2) did not differ between the 2 groups. At 2 months' evaluation, a healing scar was found in 67 patients (89%) from group I and 72 patients (96%) from group II. A relapse of the fissure was observed in 6 patients (8%) from group I who had a healing scar at 1 month, and 2 other patients never healed. A persistent fissure was present in 3 patients from group II who had no other symptoms. CONCLUSIONS: Botulinum toxin injected into the internal anal sphincter is effective in managing anal fissures and avoiding permanent complications. All patients were treated with the active drug and healed after 1 or 2 successive treatments. The results also confirm that higher doses account for a higher success rate, with little increase in complications or side effects, which is probably related to the diffusion of the toxin to the external sphincter. PMID- 11854697 TI - Intestinal anastomoses detected with a photopolymerized hydrogel. AB - BACKGROUND: This study examines the efficacy of a novel, absorbable photopolymerized hydrogel sealant, Focalseal (Focal Inc, Lexington, Mass), in protecting high-risk suture deficient intestinal anastomoses (HRA) compared with conventional sutured anastomoses (CSA). METHODS: Twenty-four New Zealand white rabbits were either randomized to small bowel HRA constructed with 4 interrupted 5/0 polyglyconate sutures and treated with Focalseal or small bowel CSA constructed with 8 to 10 interrupted 5/0 polyglyconate sutures. Four rabbits from each group were killed at postoperative days 3, 7, and 21. Anastomoses were assessed for evidence of dehiscence, adhesion formation, stenosis, and bursting pressure; they were also examined histologically for collagen content estimation and blood vessel formation. RESULTS: Mean operative time was 35 minutes (SD 5) for CSA and 35 minutes (SD 5) for HRA. There was 1 postoperative death in the HRA group. A postmortem examination revealed no evidence of anastomotic leak. The remaining 23 animals were assessed as planned. There was no evidence of anastomotic dehiscence in any animal. There was no significant difference in adhesion formation (P =.09), stenosis (P =.6), or bursting pressure (P =.2) between HRA and CSA groups. Collagen (P =.007) and blood vessel (P =.002) formation were significantly increased in HRA. CONCLUSIONS: HRA treated with Focalseal heal well and have similar strength to CSA. This technique may prove valuable in procedures such as laparoscopic bowel resection. PMID- 11854698 TI - Comparison of microsatellite instability and chromosomal instability in predicting survival of patients with T3N0 colorectal cancer. AB - BACKGROUND: At least 2 apparently independent mechanisms, microsatellite instability (MSI) and chromosomal instability, are implicated in colorectal tumorigenesis. Their respective roles in predicting clinical outcomes of patients with T3N0 colorectal cancer remain unknown. METHODS: Eighty-eight patients with a sporadic T3N0 colon or rectal adenocarcinoma were followed up for a median of 67 months. For chromosomal instability analysis, Ki-ras mutations were determined by single-strand polymerase chain reaction, and p53 protein staining was studied by immunohistochemistry. For MSI analysis, DNA was amplified by polymerase chain reaction at 7 microsatellite targets (BAT25, BAT26, D17S250, D2S123, D5S346, transforming growth factor receptor II, and BAX). RESULTS: Overall 5-year survival rate was 72%. p53 protein nuclear staining was detected in 39 patients (44%), and MSI was detected in 21 patients (24%). MSI correlated with proximal location (P <.001) and mucinous content (P <.001). In a multivariate analysis, p53 protein expression carried a significant risk of death (relative risk = 4.0, 95% CI = 1.6 to 10.1, P =.004). By comparison, MSI was not a statistically significant prognostic factor for survival in this group (relative risk = 2.2, 95% CI = 0.6 to 7.3, P =.21). CONCLUSIONS: p53 protein overexpression provides better prognostic discrimination than MSI in predicting survival of patients with T3N0 colorectal cancer. Although MSI is associated with specific clinicopathologic parameters, it did not predict overall survival in this group. Assessment of p53 protein expression by immunocytochemistry provides a simple means to identify a subset of T3N0 patients with a 4-times increased risk for death. PMID- 11854699 TI - Quercetin inhibits human vascular smooth muscle cell proliferation and migration. AB - BACKGROUND: The French paradox has been associated with regular intake of red wine, which is enriched with flavonoids. Quercetin, a flavonoid present in the human diet, exerts cardiovascular protection through its antioxidant properties. We hypothesized that the beneficial effect of quercetin also could be related to the inhibition of vascular smooth muscle cell proliferation and migration. METHODS: Human aortic smooth muscle cells (AoSMC) were grown in culture in the presence of serum. Quercetin inhibited the serum-induced proliferation of AoSMC. This inhibition was dose-dependent and not attributed to toxicity. Cell cycle analysis revealed that quercetin arrested AoSMC in the G(0)/G(1) phase. The effect of quercetin on AoSMC migration was examined using explant migration and Transwell migration assays. Quercetin significantly decreased migration in both assays in a consistent manner. Finally, Western blot analysis of AoSMC exposed to quercetin demonstrated a significant reduction in the activation of mitogen activated protein kinase, a signaling pathway associated with the migration of vascular smooth muscle cells. CONCLUSIONS: Quercetin inhibits the proliferation and migration of AoSMC, concomitant with inhibition of mitogen-activated protein kinase phosphorylation. These findings provide new insights and a rationale for the potential use of quercetin in the prophylaxis of cardiovascular diseases. PMID- 11854700 TI - It takes a faculty. PMID- 11854701 TI - Invited commentary: measure of competence. PMID- 11854702 TI - Invited commentary: comment from American Board of Surgery. PMID- 11854703 TI - Invited commentary: the ACGME "Outcomes Project". American Council for Graduate Medical Education. PMID- 11854704 TI - Sentinel lymph node biopsy demonstrating concomitant melanoma and mantle cell lymphoma. PMID- 11854705 TI - Adenovirus vector-mediated transfer of the vascular endothelial growth factor cDNA to healing abdominal fascia enhances vascularity and bursting strength in mice with normal and impaired wound healing. AB - BACKGROUND: We hypothesized that adenovirus-mediated transfer of the vascular endothelial growth factor (VEGF121) complementary DNA (cDNA) to murine laparotomy fascial wounds would enhance vascularity and bursting strength. METHODS: Microfibrillar collagen sponges saturated with adenovirus (Ad) vectors encoding for the human VEGF121 cDNA (Ad(CU)VEGF121.1), a control marker gene (Ad beta gal, AdLuc) or no transgene (AdNull) were sutured to fascial edges during laparotomy closure in normal mice and mice treated with dexamethasone. Endpoints addressed included transgene expression in the fascia and surrounding tissue, the number of blood vessels in the healing wound determined using immunostaining, and wound bursting strength using a calibrated tensinometer. RESULTS: Transgene expression was detected readily in the fascial edges, but only marginally detectable in neighboring tissues. In normal mice and mice treated with dexamethasone, no differences were observed at 7 days. Strikingly, however, 21 days after wound closure/therapy, significantly more blood vessels were present in the wounds that received the VEGF121 vector compared with controls (normal: AdNull: 4.2 +/- 1.8; Ad(CU)VEGF121.1: 11.2 +/- 1.2; P <.05; dexamethasone: AdNull: 1.4 +/- 0.8; Ad(CU)VEGF121.1: 5.4 +/- 1.2; P <.05), and bursting strength was significantly higher in VEGF121-treated wounds (normal: AdNull: 665 +/- 68 mN/mm; Ad(CU)VEGF121.1: 956 +/- 82 mN/mm; P <.0005; dexamethasone: AdNull: 234 +/- 111 mN/mm; Ad(CU)VEGF121.1: 592 +/- 121 mN/mm; P <.03). CONCLUSIONS: Adenovirus mediated gene transfer to healing fascial wounds is achieved readily using a microfibrillar collagen sponge, with transfer of the human VEGF121 cDNA significantly enhancing wound vascularity and bursting strength in normal mice, as well as in mice treated with dexamethasone. PMID- 11854706 TI - Coffee creamer, the bionic man, and organ preservation. PMID- 11854707 TI - Pseudomyxoma peritonei. PMID- 11854708 TI - Strangulated herniation through a defect in the broad ligament. PMID- 11854709 TI - Papillary cholangiocarcinoma: a cause of intermittent, obstructive jaundice. PMID- 11854710 TI - Significant hypomagnesemia after celiotomy: implications of preoperative bowel cleansing with sodium phosphate purgative. PMID- 11854711 TI - The cost of transfusion and alternatives. PMID- 11854712 TI - Eosinophilic cholangiopathy. PMID- 11854713 TI - The value of sestamibi scans. PMID- 11854714 TI - In severe jaundice, major liver resection is contraindicated. PMID- 11854715 TI - Presidential address: at the count of two. PMID- 11854716 TI - Efficacy of a bifurcated endograft versus open repair of abdominal aortic aneurysms: a reappraisal. AB - OBJECTIVE: Late complications and graft failures have recently cast serious doubts on the durability of endovascular repair of abdominal aortic aneurysms (AAA). The results of a multicenter trial comparing a bifurcated endograft (AB) with standard open repair (OR) were reviewed to assess the late findings of both methods of AAA treatment. PATIENTS AND METHODS: In a multicenter study of AB versus OR conducted from December 1995 to February 1998, 242 patients with AAA successfully treated with an AB and 111 control patients treated concurrently with OR were followed up at least yearly. Twenty-five immediate conversions were excluded from late follow-up. All imaging modalities obtained during follow-up were reviewed by a core laboratory for AAA size, endoleaks, migration, and device integrity. Clinical outcomes at the yearly visits were compared. All death reports were reviewed to classify the cause of death. RESULTS: Average follow-up for the AB group was 36 months, with 194 patients at 3 years and 55 patients at 4 years. The cumulative mortality rate was similar between the AB (15.7%) and OR groups (12.6%; P =.59). The significant early benefit to the AB group in cardiopulmonary complications was no longer evident by 3 years. However, the AB advantage in total and bowel complications, as well as the higher renal complication rates, persisted. At 3 years, 73.7% of patients showed a significant reduction of their AAA size, whereas 25.7% still had an endoleak. One migration and two single hook fractures were noted. Graftrelated reinterventions were performed in 50 patients (20%) without any deaths. Twenty-eight patients (11.6%) underwent interventions for limb flow compromise, whereas 25 were treated for endoleak. Late conversion to OR was required in five patients (2%). No AAA ruptures were encountered in either group. CONCLUSIONS: Rupture-free survival rates after treatment of AAA with the bifurcated AB are similar to those of the OR group. Notably the proximal attachment system is relatively stable and the AAA shrinks in three of four patients treated. Reinterventions are nonetheless required in nearly one of five patients. Although most late procedures are percutaneous, counseling regarding possible future interventions is necessary. PMID- 11854717 TI - Outcome of endovascular abdominal aortic aneurysm repair in patients with conditions considered unfit for an open procedure: a report on the EUROSTAR experience. AB - OBJECTIVE: Endovascular abdominal aortic aneurysm repair (EAR) can be performed in patients whose conditions were previously considered unfit for conventional treatment of the aneurysm. However, because the life span in this category of patients often is limited because of serious comorbidity, the efficacy of EAR in prolonging life expectancy remains uncertain. This study involves the evaluation of preoperative risk classification and an assessment of the outcome of interventions. METHODS: The data of 3075 patients, who underwent operation in 101 European institutions that collaborated in the EUROSTAR Registry, were assessed. Only the patients who had been prospectively enrolled in the registry were used for this analysis. Patient characteristics, operative risk factors, procedural details, and types of devices were correlated with preoperative estimates of operative risk, early and late mortality, complications, and primary and secondary outcome success rates. In addition, the intermediate-term survival rates in patients with unfit conditions with EAR (observed series) and with conservative approaches of the aneurysms (rupture rates as derived from the literature) were compared in a mathematical model. RESULTS: Of the overall study group, 2525 patients were at "normal" risk for a surgical procedure (group A), 399 patients had conditions that were considered unfit for open surgery (group B), and 151 patients had conditions that were unfit for general anesthesia (group C). Both unfit categories had significantly more comorbid factors and larger aneurysms than did the patients in good medical condition. Differences were observed in comorbidities between the two high-risk categories, groups B and C. Factors that influenced the abdominal approach (previous laparotomies, hostile abdomen, and obesity) and local anatomic factors (eg, retroperitoneal fibrosis, inflammatory aneurysm, dissections, and enterostomy) were present in 19% of the patients with conditions that were unfit for open surgery and in only 1% of the category unfit for anesthesia. In contrast, severe pulmonary disease was present in 33% of the patients with conditions that were unfit for anesthesia as opposed to 11% of the patients with conditions that were unfit for open surgery. The early and late mortality rates were significantly higher in the unfit categories (groups B and C). Life table results showed a 3-year survival rate of 83% in patients at normal operative risk and of 68% in patients with unfit conditions (P =.0001). An independent correlation with late death was shown for the clinical classification into high-risk groups B and C, pulmonary disease, team experience of less than 60 procedures, and the diameter of the aneurysm. In groups B and C, aneurysms smaller than 6.0 cm were associated with a 2-year survival rate of 80% and larger aneurysms with a rate of 68% (P =.02). This difference was caused by an increased non-aneurysm-related mortality rate in the group with aneurysms of more than 6 cm. The mathematical model showed an advantage of EAR with regard to the reduction of the death rate in patients with unfit conditions as compared with no intervention after 1 year. The advantage of EAR was observed in patients with AAAs between 5 and 6 cm and with larger aneurysms. CONCLUSION: Early and late mortality rates were increased in patients with the preoperative clinical diagnosis "unfit for open surgery and general anesthesia" as compared with patients at "normal" operative risk. EAR appeared of potential benefit in patients with unfit conditions, regardless of the aneurysm diameter. The life expectancy of patients at high risk who are considered for EAR should be longer than 1 year before any realistic gain in life span can be anticipated. PMID- 11854718 TI - Durability of benefits of endovascular versus conventional abdominal aortic aneurysm repair. AB - PURPOSE: Endovascular abdominal aortic aneurysm (AAA) repair is reported to result in less initial patient morbidity and a shorter hospital length of stay (LOS) when compared with conventional AAA repair. We sought to examine the durability of this result during the intermediate follow-up interval. METHODS: The records of all admissions for all patients who underwent AAA repair during a 26-month interval were reviewed. RESULTS: Three hundred thirty-seven (337) patients underwent procedures to repair AAAs (163 open and 174 endovascular). Endovascular procedures were performed with a variety of devices (Talent, 108; Ancure, 36; AneuRx, 26; Zenith, 2; and Cordis, 2) and configurations (141 bifurcated and 33 aortomonoiliac). The mean follow-up period was 10.6 months (endovascular repair) and 12.3 months (open repair). LOS did not significantly vary by device (P =.24 to P =.92) or configuration (P =.24). The initial median LOS for procedures was significantly shorter (P =.009) for endovascular repairs (5 days) than for open procedures (8 days). However, the patients who underwent endovascular repair were more likely to be readmitted during the follow-up interval when compared with patients who underwent open procedure. The readmission-free survival rate after AAA repair at 12 months was 95% for patients for open AAA repair versus 71% for patients for endovascular repair (P <.001). If the total hospital days were compared, including the initial and all subsequent AAA-related admissions, there was no significant difference for mean LOS for patients who underwent endovascular versus open AAA procedures (11 days versus 13.6 days; P =.21). The patients for endovascular AAA repair most commonly needed readmission for treatment of endoleak (n = 31), wound infection (n = 12), and graft limb thrombosis (n = 9). Although women had similar LOS to men for endovascular repair (P =.44), they had longer initial LOS for open AAA repair (15 versus 10 days; P =.03). After endovascular repair, women were more likely than men to be readmitted by 12 months (51% versus 71% readmission-free survival rate; P =.03) and they had longer LOS on readmission (13.2 versus 5.2 days; P =.006). No gender differences were identified for patients after open AAA repair regarding readmission-free survival rate (P =.09) or LOS on readmission (P =.98). CONCLUSION: Although initial LOS was shorter for the patients who underwent endovascular as compared with conventional AAA repair, this advantage was lost during the follow-up interval because of frequent readmission for the treatment of procedure-related complications, chiefly endoleak. These readmissions frequently involved the performance of additional invasive procedures. Gender differences existed regarding LOS and the likelihood of complications after open and endovascular AAA repair. PMID- 11854719 TI - Device migration after endoluminal abdominal aortic aneurysm repair: analysis of 113 cases with a minimum follow-up period of 2 years. AB - PURPOSE: Device migration (DM) has been shown to cause late failure after endoluminal abdominal aortic aneurysm (AAA) repair. To establish the incidence rate and the predictive factors of distal migration of the proximal portion of the endograft, computed tomographic (CT) scans performed at different time intervals during follow-up examination of 113 patients were reviewed. PATIENTS AND METHODS: Between April 1997 and March 1999, 148 patients underwent endoluminal AAA repair with a modular endograft with infrarenal fixation (Medtronic-AVE AneuRx, Santa Rosa, Calif) at our unit. CT scans performed at 1, 6, and 12 months after surgery and yearly thereafter were prospectively stored in a computer imaging database. Patient demographics, risk factors, operative details, and follow-up events were prospectively collected. No patients were lost to follow-up examination. Twelve patients died within 2 years of surgery, four patients underwent immediate conversion to open repair, and adequate CT measurements were not feasible in 19 cases, which left 113 patients available for a minimum 2-year assessment and 418 CT scan results reviewed. Two vascular surgeons, blinded to patient identity and history with tested interobserver agreement (kappa = 0.64), separately reviewed axial reconstructions of CT scans. DM was defined as changes of 10 mm or more in the distance between the lower renal artery and the first visible portion of the endograft at follow-up examination. Ten possible independent predictors of DM were analyzed with multivariate Cox proportional hazards regression model. RESULTS: One AAA rupture, which was successfully treated, occurred at a mean follow-up period of 28 months (range, 24 to 46 months). Seventeen patients (15%) showed DM. Eight patients (47%) with DM underwent reintervention: a proximal cuff was positioned in six patients and late conversion to open repair was performed in two patients. Of the 10 variables analyzed with Cox proportional hazards regression model, AAA neck enlargement of more than 10% after endoluminal repair (hazard ratio, 7.3; confidence interval, 1.8 to 29.2; P =.004) and preoperative AAA diameter of 55 mm or more (hazard ratio, 4.5; confidence interval, 1.2 to 16.7; P =.02) were positive independent predictors of DM. The probability of DM at 36 months was 27% according to life table analysis. CONCLUSION: DM occurred in a significant portion of our patients, yet aggressive follow-up examination and a high reintervention rate prevented aneurysm-related death. According to our data, dilatation of the infrarenal aortic neck is an important factor that contributes to the distal migration of stent grafts, and patients with large aneurysms are at high risk for DM. PMID- 11854720 TI - Surgical management of atherosclerotic renovascular disease. AB - OBJECTIVE: This review describes the clinical outcome of surgical intervention for atherosclerotic renovascular disease in 500 consecutive patients with hypertension. METHODS: From January 1987 to December 1999, 626 patients underwent operative renal artery (RA) repair at our center. A subgroup of 500 patients (254 women and 246 men; mean age, 65 plus minus 9 years) with hypertension (mean blood pressure, 200 plus minus 35/104 plus minus 21 mm Hg) and atherosclerotic RA disease forms the basis of this report. Hypertension response was determined from preoperative and postoperative blood pressure measurements and medication requirements. Change in renal function was determined with estimated glomerular filtration rates (EGFRs) calculated from serum creatinine levels. Proportional hazards regression models were used for the examination of associations between selected preoperative parameters, blood pressure and renal function response, and eventual dialysis-dependence or death. RESULTS: Two hundred three patients underwent unilateral RA procedures, 297 underwent bilateral RA procedures, and 205 patients underwent combined renal and aortic reconstruction. After surgery, there were 23 deaths (4.6%) in the hospital or within 30 days of surgery. Significant and independent predictors of perioperative death included advanced age (P <.0001; hazard ratio [HR], 3.23; 95% confidence interval [CI], 1.85 to 5.70) and clinical congestive heart failure (P =.013; HR, 3.05; 95% CI, 1.26 to 7.34). Among the patients who survived surgery, hypertension was considered cured in 12%, improved in 73%, and unchanged in 15%. For the entire group, renal function increased significantly after operation (preoperative versus postoperative mean EGFR, 41.1 plus minus 23.9 versus 48.2 plus minus 25.5 mL/min/m(2); P <.0001). For individual patients, with a 20% or more change in EGFR considered significant, 43% had improved renal function (including 28 patients who were removed from dialysis-dependence), 47% had unchanged function, and 10% had worsened function. Preoperative renal insufficiency (P <.001; HR, 2.35; 95% CI, 1.86 to 2.98), diabetes mellitus (P =.007; HR, 2.14; 95% CI, 1.15 to 3.97), prior stroke (P =.042; HR, 1.50; 95% CI, 1.02 to 2.22), and severe aortic occlusive disease (P =.003; HR, 1.69; 95% CI, 1.19 to 2.31) showed significant and independent associations with death or dialysis during the follow up examination period. After operation, blood pressure cured (P =.014; HR, 0.52; 95% CI, 0.30 to 0.88) and improved renal function (P =.011; HR, 0.40; 95% CI, 0.19 to 0.81) showed significant and independent associations with improved dialysis-free survival rate. All categories of function response and time to death or dialysis showed significant interactions with preoperative EGFR. CONCLUSION: The surgical correction of atherosclerotic renovascular disease resulted in blood pressure benefit and retrieval of renal function in selected patients with hypertension. The patients with cured hypertension or improved EGFR after operation showed increased dialysis-free survival as compared with other patients who underwent surgery. PMID- 11854721 TI - Early abdominal closure with mesh reduces multiple organ failure after ruptured abdominal aortic aneurysm repair: guidelines from a 10-year case-control study. AB - OBJECTIVE: The objectives of this study were the comparison of patients who needed mesh closure of the abdomen with patients who underwent standard abdominal closure after ruptured abdominal aortic aneurysm repair and the determination of the impact of timing of mesh closure on multiple organ failure (MOF) and mortality. METHODS: We performed a case-control study of patients who needed mesh based abdominal closure (n = 45) as compared with patients who underwent primary closure (n = 90) after ruptured abdominal aortic aneurysm repair. RESULTS: Before surgery, the patients who needed mesh abdominal closure had more blood loss (8 g versus 12 g of hemoglobin; P <.05), had prolonged hypotension (18 minutes versus 3 minutes; P <.01), and more frequently needed cardiopulmonary resuscitation (31% versus 2%; P <.01) than did the patients who underwent primary closure. During surgery, the patients who needed mesh closure also had more severe acidosis (base deficit, 14 versus 7; P <.01), had profound hypothermia (32 degrees C versus 35 degrees C; P <.01), and needed more fluid resuscitation (4.0 L/h versus 2.7 L/h; P <.01). With this adverse clinical profile, the patients who needed mesh closure had a higher mortality rate than did the patients who underwent primary closure (56% versus 9%; P <.01). However, the patients who underwent mesh closure at the initial operation (n = 35) had lower MOF scores (P <.05), a lower mortality rate (51% versus 70%), and were less likely to die from MOF (11% versus 70%; P <.05) than the patients who underwent mesh closure after a second operation in the postoperative period for abdominal compartment syndrome (n = 10). CONCLUSION: This study reports the largest experience of mesh-based abdominal closure after ruptured abdominal aortic aneurysm repair and defines clinical predictors for patients who need to undergo this technique. Recognition of these predictors and initial use of mesh closure minimize abdominal compartment syndrome and reduce the rate of mortality as the result of MOF. PMID- 11854722 TI - Monocyte chemotactic activity in human abdominal aortic aneurysms: role of elastin degradation peptides and the 67-kD cell surface elastin receptor. AB - BACKGROUND: Chronic inflammation is a characteristic feature of abdominal aortic aneurysms (AAAs), but the molecular signals responsible for recruiting monocytes into the outer aortic wall are unresolved. The purpose of this study was to examine whether AAA tissues elaborate chemotactic activity for mononuclear phagocytes and to determine whether this activity is attributable to interactions between elastin degradation peptides (EDPs) and their cell surface receptor, the 67-kD elastin binding protein (EBP). MATERIAL AND METHODS: Soluble proteins were extracted from human AAA tissues, and chemotactic activity for differentiated U937 mononuclear phagocytes was measured by use of a modified Boyden chamber. Chemotactic activity induced by N -formyl-Met-Leu-Phe was used as a positive control and checkerboard analysis was used to distinguish chemotaxis from chemokinesis. Inhibition of chemotaxis was tested by peptide competition, blocking antibodies and galactosugar-mediated dissociation of the 67-kD EBP. RESULTS: AAA extracts stimulated a concentration-dependent increase in monocyte migration that reached up to 24% of the maximal effect induced by N -formyl-Met Leu-Phe. Checkerboard analysis demonstrated that AAA extracts stimulated chemotaxis without a chemokinetic effect. AAA-derived chemotactic activity was eliminated by competition with Val-Gly-Val-Arg-Pro-Gly (VGVAPG), a repetitive peptide found in human elastin that binds to cellular elastin receptors, and decreased nearly 40% in the presence of BA-4, an antielastin monoclonal antibody that can block EDP-mediated chemotactic activity. Monocyte chemotaxis in response to both VGVAPG and AAA extracts was abolished in the presence of lactose, a galactosugar that specifically dissociates the 67-kD EBP, but it was unaffected by either glucose, fructose, or mannose. CONCLUSIONS: These findings indicate that soluble EDPs released within human AAA tissue can subsequently attract mononuclear phagocytes through ligand-receptor interactions with the 67-kD EBP, thereby providing a plausible molecular mechanism to explain the inflammatory response that accompanies aneurysmal degeneration. Better understanding of factors regulating inflammatory cell recruitment may lead to novel forms of therapy for early stages of aneurysmal degeneration. PMID- 11854723 TI - Spinal cord arteriography: a safe adjunct before descending thoracic or thoracoabdominal aortic aneurysmectomy. AB - OBJECTIVE: Spinal cord arteriography (SCA) often has been considered difficult, hazardous, and unreliable. In this report, we question these assumptions. PATIENTS: From August 1985 to June 2000, a total of 480 patients underwent 487 SCA procedures during diagnostic examination for 498 aneurysms, which included 159 that involved the descending thoracic aorta and 339 that involved the thoracoabdominal aorta. The underlying cause was degenerative disease in 288 cases, chronic dissection in 132 cases, and other causes in 78 cases. RESULTS: Major procedure-related complications occurred in six patients (1.2%) and included spinal cord complications in two patients, renal complications in two patients, and stroke in two patients. Puncture-site complications occurred in three patients (0.6%). Rupture of the aneurysm occurred within 3 days after SCA in two patients (0.4%). Two deaths (0.4%) were directly imputable to SCA. In 476 patients (97.7%), SCA was devoid of major complications. The Adamkiewicz's artery was successfully located in 419 patients (86.0%) and arose from a left intercostal or lumbar artery in 323 patients (77.1%) and from between T8 and L1 levels in 361 patients (86.2%). On the basis of the extent of identification of spinal cord vasculature, the procedure was considered as a complete success in 321 patients (65.9%), as a partial success in 112 patients (23.0%), and as a failure in 54 patients (11.1%). Although the failure rates were comparable, the complete success rate was significantly higher in patients with degenerative rather than dissecting aneurysms (P <.001) and in patients with limited aneurysms (ie, types 1, 2, and 3 versus type 4 descending thoracic aneurysms, P <.05; and types 3 and 4 versus types 1 and 2 thoracoabdominal aneurysms, P <.001). CONCLUSION: SCA is a safe adjunct that warrants more widespread use in the management of descending thoracic or thoracoabdominal aortic aneurysms. PMID- 11854724 TI - Surgical sympathectomy for reflex sympathetic dystrophy syndromes. AB - PURPOSE: The purpose of this study was the assessment of the efficacy of thoracoscopic cervicodorsal and open lumbar sympathectomy for the reduction of pain severity and disability associated with reflex sympathetic dystrophy (RSD). METHODS: From 1992 to 2000, 73 patients with RSD underwent 46 video-assisted thoracoscopic (first to fourth thoracic ganglion) or 37 surgical lumbar (first to fourth lumbar ganglion) sympathetic chain resections. The patients were referred from multidisciplinary pain clinics with documented sympathetically maintained pain syndrome on the basis of reproducible more than 50% reduction in pain severity score (0, no pain; 10, most severe pain imaginable) for more than 2 days after sympathetic block therapy. The mean duration of the RSD symptoms before sympathectomy was 26 plus minus 14 months (range, 6 to 100 months). Postoperative pain severity score, limb disability, and overall patient satisfaction were assessed by an independent third-party observer at a mean follow-up period of 30 months. RESULTS: No operative mortality or serious morbidity (Horner's syndrome, bleeding that needed transfusion, wound infection) occurred. Transient (<3-month) postprocedural sympathalgia developed in one third of the patients for cervicodorsal sympathectomy and 20% of the patients for lumbar sympathectomy and was treated effectively with trigger point/proximal ganglion block therapy or transcutaneous electrical nerve stimulation. At 3 months after sympathectomy, 10% of the patients had conditions that were judged treatment failures with no reduction in pain severity or limb disability. The remaining patients testified to more than 50% pain reduction, with pain severity scores decreasing from a mean of 8.7 before surgery to 3.4 after sympathectomy. At 1 year, one quarter of the patients had continued significant pain relief (pain severity score, <3) and an additional 50% of the patients indicated continued but reduced pain severity and an increase in daily/work activities. Overall, patient satisfaction (willingness to have procedure again, benefit from sympathectomy) was 77% and was not significantly influenced by patient age, RSD duration/stage, or extremity involvement (lumbar, 84%; cervicodorsal, 72%). CONCLUSION: Patients with RSD with a confirmed sympathetically maintained pain syndrome can realize long-term benefit from surgical sympathectomy. Procedural efficacy was similar for both upper limb and lower limb RSD syndromes, although the level of pain reduction did deteriorate with time. After sympathectomy, the patients with RSD had a low incidence rate (7%) of "new" complex regional pain or disabling compensatory sweating syndromes. PMID- 11854725 TI - Coagulation, fibrinolysis, and recanalization after acute deep venous thrombosis. AB - OBJECTIVE: Recanalization is common after acute deep venous thrombosis, but the factors that contribute to its variable extent are unknown. The purpose of this study was to examine the relationship between recanalization and plasma markers of coagulation and fibrinolysis. METHODS: Subjects with an ultrasound-confirmed deep venous thrombosis had prothrombin fragment 1+2 (F 1+2), tissue plasminogen activator (t-PA) activity, plasminogen activator inhibitor (PAI-1) activity, and t-PA antigen levels determined before anticoagulation therapy. Ultrasound and plasma studies were repeated at 14 days, 1 month, and every 3 months for 1 year. RESULTS: Among 71 enrolled subjects, F 1+2 levels normalized within 14 days. The mean t-PA activity was within the normal range at all follow-up intervals. However, the mean t-PA antigen (10.7 plus minus 10.5 to 13.6 plus minus 13.5 ng/mL; P =.04) and PAI-1 (9.0 plus minus 8.1 to 13.2 plus minus 17.3 U/mL; P =.05) levels increased between the time of presentation and day 14. The mean reduction in thrombus score among 44 patients who completed 9 months of follow-up was 60.9% (plus minus 42.1%). Percent recanalization was directly associated with initial t-PA activity levels (R =.4; P =.006) and inversely related to F 1+2 (R = minus sign. 5; P =.004), t-PA antigen (R = minus sign.5, P =.002), and PAI-1 (r = minus sign. 5, P =.001) levels. However, only initial F 1+2 (P =.0009) and t-PA antigen (P =.004) levels were independent predictors of the degree of recanalization. CONCLUSION: Although the mechanisms by which the venous lumen is restored are still being elucidated, recanalization is inversely related to levels of activated coagulation (F 1+2) and fibrinolytic inhibition (t-PA antigen) at the time of presentation. PMID- 11854726 TI - Duplex scan-directed placement of inferior vena cava filters: a five-year institutional experience. AB - PURPOSE: The purpose of this study was the assessment of the safety, efficacy, and hospital charges of bedside duplex ultrasound-directed inferior vena cava (IVC) filter placement. METHODS: All duplex ultrasound-directed IVC filters that were placed from August 8, 1995, to December 31, 2000, are reviewed. Chart review combined with mailed questionnaires and telephone follow-up examinations were used to collect demographic and outcome data. RESULTS: Three hundred twenty-five patients underwent evaluation, and 284 underwent duplex ultrasound-directed IVC filter placement. Two hundred three (71%) were male patients, and 81 (29%) were female patients. Poor IVC visualization, IVC thrombosis, and unsuitable anatomy prevented duplex-directed filter placement in 41 patients (12%). Indication for filter placement included venous prophylaxis in the absence of thromboembolism in 235 patients (83%), contraindication to anticoagulation therapy in 34 patients (12%), prophylaxis with therapeutic anticoagulation therapy in the presence of thromboembolism in 7 patients (2%), and complication of anticoagulation therapy in 8 patients (3%). There were no procedure-related deaths or septic complications. Technical complications occurred in 12 patients (4%). Filter misplacement occurred in 6 patients (2%), access thrombosis in 1 (<1%), migration in 1 (<1%), bleeding in 1 (<1%), and IVC occlusion in 3 (1%). Pulmonary emboli after IVC filter placement occurred in one patient with a misplaced filter. Average hospital charges related to duplex ultrasound-directed filter placement were $2388 less than fluoroscopic placement charges in the year 2000. CONCLUSION: Our experience indicates that duplex ultrasound-directed IVC filter placement is safe, cost-effective, and convenient for patients who need IVC filter placement. PMID- 11854727 TI - The effect of anticoagulation therapy and graft selection on the ischemic consequences of femoropopliteal bypass graft occlusion: results from a multicenter randomized clinical trial. AB - OBJECTIVE: A recent retrospective study showed that the ischemic consequences of femoropopliteal bypass graft occlusion were more severe with polytetrafluoroethylene (PTFE) than with vein. This study examines this conclusion and whether oral anticoagulation therapy reduces the degree of ischemia after occlusion of PTFE and vein femoropopliteal bypass grafts. METHODS: Four hundred two patients who underwent femoropopliteal bypass grafting (233 PTFE and 169 vein) were randomized to a postoperative regimen of either warfarin (international normalized ratio, 1.4 to 2.8) and aspirin (WASA; 325 mg daily) therapy or aspirin alone (ASA) therapy. The grade of acute ischemia at the time of graft occlusion was assessed with the Society of Vascular Surgery recommended reporting standards (I, viable; II, threatened). Early graft occlusions (<30 days) were excluded. RESULTS: There were 100 graft occlusions (67 PTFE and 33 vein) during a mean follow-up period of 36 months (PTFE) and 39 months (vein). Forty-eight patients were randomized to WASA therapy, and 52 were randomized to ASA therapy. The patients were well matched for age, atherosclerotic risk factors, operative indication, and preoperative ankle-brachial index. Overall, a greater percentage of the PTFE occlusions caused grade II ischemia than did the vein graft occlusions (48% versus 18%; P =.005). The ankle-brachial index at the time of graft occlusion was significantly lower in the PTFE grafts than in the vein grafts (0.28 versus 0.45; P =.001). The patients with PTFE who were undergoing WASA therapy at the time of graft occlusion had less grade II ischemia than did those patients who were undergoing ASA therapy (28% versus 55%; P =.057). However, the incidence rate of severe ischemia after graft occlusion remained greater with PTFE grafts and WASA therapy as compared with all the vein grafts (28% versus 18%). The vein graft occlusions had the same incidence rate of grade II ischemia with WASA therapy as with ASA therapy (20% versus 17%; P = 1.0). CONCLUSION: The ischemic consequences of femoropopliteal bypass graft occlusion are worse with PTFE than with vein. Treatment with WASA therapy lessens the severity of acute ischemia after the occlusion of PTFE graft as compared with ASA therapy but not to the degree seen with vein graft occlusion. Occlusion of femoropopliteal vein grafts is seldom accompanied by severe ischemia and is not improved with WASA therapy. PMID- 11854728 TI - Early results of a prospective randomized trial of spliced vein versus polytetrafluoroethylene graft with a distal vein cuff for limb-threatening ischemia. AB - OBJECTIVE: Single-piece vein remains the conduit of choice in patients who need bypass grafting for limb salvage. When this option is not available, two of the remaining options are prosthetic bypass graft or several segments of vein spliced together. In this study, we compare spliced vein bypass grafting versus polytetrafluoroethylene grafting with a distal vein cuff in patients with limb threatening ischemia. METHODS: Between 1996 and 2000, 39 bypass grafting procedures in 36 patients were performed for limb-threatening ischemia. These procedures were prospectively randomized to either spliced vein bypass grafting (spliced group, 19 bypass grafts) or polytetrafluoroethylene grafting with a distal vein cuff (cuff group, 20 bypass grafts). All the patients in the cuff group underwent anticoagulation therapy with warfarin sodium after surgery. The inclusion criteria included: no single-piece vein option for bypass grafting, adequate vein for splice, no composite sequential option, and limb-threatening ischemia. The demographics were similar between the two groups. RESULTS: The primary patency rate at 2 years was 44% and 49% for the spliced and cuff groups, respectively. In the spliced group, seven of 19 bypass grafts underwent revision in the follow-up period, and two of 20 cuffed bypass grafts were successfully revised. The secondary patency rate was 87% and 59% (P <.05), with limb salvage rates of 94% and 85% for spliced and cuff groups, respectively. Four patients in the spliced vein group needed reoperation for wound complications related to vein harvest. One polytetrafluoroethylene graft needed removal for infection. Two early mortalities occurred in the spliced group, one from myocardial infarction and one from stroke. The overall survival rate at 2 years between the two groups was 67% and 100% for the spliced and cuff groups, respectively (P <.05). CONCLUSION: Although this is a preliminary report, it appears that both spliced vein bypass grafting and polytetrafluoroethylene bypass grafting with a distal vein cuff produce acceptable limb salvage rates. The secondary patency rate for spliced vein is better, but these bypass grafts more often need revision or reoperation for wound complications. PMID- 11854729 TI - Lysis/balloon angioplasty versus thrombectomy/ open patch angioplasty of failed femoropopliteal polytetrafluoroethylene bypass grafts. AB - PURPOSE: Several studies have reported on the outcome of lysis/percutaneous transluminal balloon angioplasty (PTA) of failed or failing femoropopliteal bypass grafts (FPGs) with mixed results. None of these studies have compared the results of lysis/PTA versus thrombectomy/open patch repair for failed above-knee polytetrafluoroethylene (PTFE) FPGs. METHODS: Patients with failed (thrombosed) above-knee FPGs (PTFE, Goretex) during a 10-year period were given the option to choose between thrombectomy/open patch repair for localized anastomotic short stenosis (less-than-or-equal2 cm; group A, 31 patients) and lysis/PTA or thrombectomy/balloon angioplasty when lysis failed or was contraindicated (group B, 26 patients). The cumulative patency rates were compared by using a Kaplan Meier life table analysis. All patients underwent routine color duplex ultrasound scanning/ankle brachial index measurements at 30 days, 6 months, and every 6 months thereafter. RESULTS: Demographic and clinical characteristics and indications for intervention were comparable in both groups. The mean follow-up period was 54.1 and 46.2 months in group A and group B, respectively. There were four perioperative complications in group A (13%) and seven perioperative complications in group B (27%). Initial technical success and 30-day secondary graft patency rates were 100% in both groups. Overall, 17 of 31 patients (55%) had open grafts, with no further revisions in group A and six of 26 patients (23%) in group B requiring further revisions (P =.012). Nine of 31 grafts (29%) failed in group A versus 15 of 26 grafts (58%) in group B (P =.027). The rate of limb loss was comparable in both groups (6% vs 12%). The overall cumulative secondary patency rates at 6 months and 1, 2, 3, 4, and 5 years were 100%, 93%, 85%, 72%, 67%, and 62% for group A and 100%, 96%, 88%, 76%, 63%, and 45% for group B (P =.035). Thirty-five further interventions were needed to maintain graft patency in group B (mean, 1.35; range, 0-3) versus five further interventions in group A (mean, 0.16; range, 0-1; P <.05). CONCLUSION: Thrombectomy/open surgical repair is superior to lysis/PTA (or thrombectomy/balloon angioplasty) for the treatment of failed above-knee PTFE FPGs with anastomotic stenoses. Therefore, balloon angioplasty should be reserved for patients who are at high risk for surgery. PMID- 11854730 TI - Intravascular ultrasound scanning improves long-term patency of iliac lesions treated with balloon angioplasty and primary stenting. AB - PURPOSE: Underdeployment of an intravascular stent has been identified as a cause of restenosis or occlusion of a treated arterial lesion. Intravascular ultrasound (IVUS) has been shown to initially improve the anatomic and clinical stenting. The purpose of this study was to determine whether the use of IVUS increased long term patency of this intervention. METHODS: Between March 1992 and October 1995, 71 limbs (52 patients) with symptomatic aortoiliac occlusive disease underwent balloon angioplasty with primary stenting. IVUS and arteriography were used in 49 limbs (36 patients) to evaluate stent deployment. Arteriography alone was used in 22 limbs (16 patients) to evaluate stent deployment. Patients were captured prospectively in a vascular registry and retrospectively reviewed. RESULTS: Mean age of patients treated with IVUS was 61.1 plus minus 9.0 years (range, 38-85) versus 70.0 plus minus 10.1 years (range, 57-87) in patients treated without IVUS (P <.01). There was no difference between the groups with respect to preoperative comorbidities, ankle-brachial indices, or number of stents per limb. Mean follow up for IVUS patients was 62.1 plus minus 7.3 months (range, 15-81) and 57.9 plus minus 8.7 months (range, 8-80) for patients treated without IVUS (P = not significant). In 40% (20/49) of limbs, IVUS demonstrated inadequate stent deployment at the time of the original procedure. Kaplan-Meier 3- and 6-year primary patency estimates were 100% and 100% in the IVUS group and 82% and 69%, respectively, in limbs treated without IVUS (P <.001). There have been no secondary procedures performed in limbs treated with IVUS and a 23% (5/22) secondary intervention rate in the non-IVUS group (P <.05). Overall Kaplan-Meier survival estimates at 3 and 6 years for all patients were 84% and 67%, respectively. CONCLUSION: Balloon angioplasty and primary stenting of symptomatic aortoiliac occlusive lesions is a durable treatment option. Long-term follow-up of treated patients shows outcomes that are comparable with direct surgical intervention. IVUS significantly improved the long-term patency of iliac arterial lesions treated with balloon angioplasty and stenting by defining the appropriate angioplasty diameter endpoint and adequacy of stent deployment. PMID- 11854731 TI - A prospective, randomized study of saphenous vein patching versus synthetic patching during carotid endarterectomy. AB - OBJECTIVE: The objective of this study was the determination of whether the choice of either autogenous saphenous vein (ASV) or synthetic material for patch angioplasty significantly influences the results after carotid endarterectomy (CEA). METHODS: With Institutional Review Board approval, 195 patients (145 men and 50 women; mean age, 69 years) who underwent 207 CEAs were prospectively randomized to arteriotomy closure with ASV or synthetic patches from July 1996 to January 2000. One hundred and one patients (52%) were randomized to the ASV cohort, and 94 (48%) were randomized to the synthetic cohort. Aside from a slight gender imbalance (70% versus 79% male in the ASV versus the synthetic group), there were no clinically important differences in baseline demographic variables, risk factors, or surgical indications between the ASV and synthetic groups. RESULTS: With all 207 randomized procedures on an intent-to-treat basis, there were two early (<30 days) postoperative deaths (1%). There were three perioperative strokes in the ASV cohort (3.0%) and two in the synthetic cohort (2.1%; P =.99). Two of these early strokes occurred in a subset of nine patients who received neither patch material, all after randomization but before CEA. Two patients in each group had late strokes. The cumulative freedom from stroke rate at 1 year (ASV, 94%; synthetic, 95%) was virtually identical for both cohorts. With the 125 patients who had at least one postoperative duplex scan, the incidence rate of recurrent (>or=60%) carotid stenosis was 4.8% (three of 62) for the ASV group and 6.3% (four of 63) for the synthetic group (P =.99). CONCLUSION: No significant differences in the stroke, mortality, or restenosis rates were shown between the ASV and the synthetic cohorts. While conceding the power limitations inherent in this study, we conclude that CEA may be safely performed with similar early results with ASV or synthetic patches. PMID- 11854732 TI - Hemodynamic benefits of regional anesthesia for carotid endarterectomy. AB - OBJECTIVE: The objective of this study was to define differences in perioperative hemodynamics and associated outcomes in patients who undergo carotid endarterectomy (CEA) with regional and general anesthesia. METHODS: All the patients who underwent CEA during a 25-month period were reviewed, with a comparison of those who underwent operation with cervical block anesthesia (CB) with those who underwent operation with general anesthetic (GA). Baseline intraoperative and postoperative blood pressure and heart rate were recorded, and deviation from initial values was calculated. The administration of vasoactive medications was assessed. Operative time, intensive care unit admission, postoperative length of stay, and cardiac/neurologic morbidity were recorded. RESULTS: From October 16, 1998, to December 1, 2000, 550 nonemergent CEAs were performed in 527 patients (226 with CB and 324 with GA). The patients in both groups were similar in age, presentation, and associated comorbidities. Although baseline blood pressure and heart rates were similar in both groups, those patients who underwent operation with GA had significantly greater intraoperative and postoperative hemodynamic variability and received more vasoactive medications during surgery (87% versus 51%; P <.001) and in the recovery room (36% versus 21%; P =.0009). Major postoperative blood pressure derangements were more common in the GA group (18% versus 10%; P <.05). Patients who underwent operation with GA more frequently needed intensive care unit admission (16% versus 7%; P =.01) and had more frequent delays in discharge (20% versus 11%; P =.008; postoperative length of stay, 2.1 versus 1.6 days; P =.01). Although no difference was seen in neurologic morbidity rates between groups (combined major stroke/death rate, 1.8%), the major cardiac morbidity rate was noted to be lower in the CB group (1% versus 4%; P =.05). The total in-room time was shorter in the CB group (108 versus 122 minutes; P <.001). CONCLUSION: CEA performed with CB is associated with significantly less perioperative hemodynamic instability than with GA. This results in fewer major adverse cardiac events. Ultimately, decreased critical care resource use is realized as is a shortened length of stay. PMID- 11854733 TI - Pseudo-occlusions of the internal carotid artery: a rationale for treatment on the basis of a modified carotid duplex scan protocol. AB - PURPOSE: We report on a modified duplex scanning technique that may be a means of detecting a patent internal carotid artery (ICA) previously believed to be occluded by means of magnetic resonance angiography (MRA), standard duplex protocols, or both. In addition, we attempted to develop selection criteria for operability in this setting, on the basis of the lumen diameter and wall thickness of the post-stenotic ICA segment. METHOD: In the past 22 months, 17 patients (12 men; 5 women) with ICA occlusions reported by means of MRA (10 patients) or by means of duplex scanning (7 patients) were found to have patent arteries when subjected to this duplex scanning protocol: (1) the use of low pulse repetition frequency (150-350 Hz), maximal persistence, and sensitivity of color and power angiography modes; (2) the use of an 8-MHz to 5-MHz probe as a means of visualizing the most distal extracranial segment of the ICA; and (3) measurements of the lumen diameter and wall thickness of the post-stenotic ICA. The age of patients ranged from 53 to 80 years (mean age, 71 years). Seven patients (41%) had no symptoms. RESULTS: Extremely low peak systolic and end diastolic velocities were detected distal to the stenotic segment in the ICA in all cases, and they varied from 5 to 30 cm/s (mean, 14 plus minus 8 cm/s) and 0 to 8 cm/s (mean, 4.5 plus minus 2.0 cm/s), respectively. The luminal diameter of the post-stenotic ICA varied from 0.7 to 3.6 mm (mean, 2.0 plus minus 1.1 mm), and the wall thickness ranged from 0.6 to 1.4 mm (mean, 0.9 plus minus 0.3 mm) in all patients. Twelve patients (71%) were examined with the intent of performing an endarterectomy. Of these, eight patients (47%) underwent successful operations with patches (3 vein; 5 synthetic), and four (29%) were found to have unreconstructable disease. The ICA lumen diameter and wall thickness in all eight patients who underwent endarterectomies were 2 mm or larger and 1 mm or thinner, respectively, whereas they were smaller than 2 mm and thicker than 1 mm, respectively, in the remaining four patients (P <.01). The last five patients were observed because they had small ICAs (lumen <2 mm) with thickened walls (>1 mm). Intraoperative and early postoperative duplex scanning examinations were performed in the eight ICAs that were successfully reconstructed. In these patients, the ICA lumen diameter increased from a mean of 2.9 plus minus 0.4 mm preoperatively to a mean of 4.4 plus minus 0.3 mm 2 weeks postoperatively (P <.001). Intraoperative ICA flow volumes were also measured after the endarterectomy, and they varied from 55 to 242 mL/min (mean, 115 plus minus 53 mL/min) and ranged from 122 to 220 mL/min (mean, 159 plus minus 34 mL/min) 2 weeks postoperatively. One patient who did not undergo surgical exploration died of chronic renal failure and congestive heart failure within the first month of follow-up. The remaining 16 patients had no neurological symptoms and were alive after a follow-up period of 2 to 22 months (mean, 8 plus minus 5 months). CONCLUSION: The proposed duplex protocol appears to be an effective means of identifying some patients with patent ICAs that were believed to be occluded by means of standard examinations. In addition, such patients may be candidates for an endarterectomy if the ICA post-stenotic lumen diameter is 2 mm or larger and the wall thickness is 1 mm or thinner. PMID- 11854735 TI - Localization of nitric oxide synthase in saphenous vein grafts harvested with a novel "no-touch" technique: potential role of nitric oxide contribution to improved early graft patency rates. AB - OBJECTIVE: The use of the saphenous vein in coronary artery bypass graft surgery is associated with high 1-year occlusion rates of as much as 30%. A new "no touch" technique of saphenous vein harvesting in which the vein is harvested with a pedicle of surrounding tissue and not distended may result in improved early patency rates. We hypothesize that nitric oxide synthase is better preserved with the no-touch technique, and the aim of this study was the investigation of whether nitric oxide synthase distribution and quantity in saphenous veins harvested with the no-touch technique differ from those veins harvested with the conventional technique. The separate contribution of perivascular tissue removal and distension to alterations in nitric oxide synthase was also studied. METHODS: Segments of 10 saphenous veins were harvested from 10 patients who underwent coronary artery bypass grafting surgery with the no-touch and conventional techniques. Samples were also taken from segments that were stripped of surrounding tissue but not distended. Nitric oxide synthase distribution was studied with reduced nicotinamide adenine dinucleotide phosphate--diaphorase histochemistry, and staining was quantified with image analysis. Immunohistochemistry was used for the identification of specific nitric oxide synthase isoforms, and immunomarkers were used for the identification of associated cell types. RESULTS: Nitric oxide synthase content was higher in no touch vessels as compared with conventionally harvested vessels (35.5%; P <.05, with analysis of variance). This content was associated with endothelial nitric oxide synthase on the lumen while all three isoforms were present in the media. In the intact adventitia of no-touch vessels, all three isoforms of nitric oxide synthase were also present, associated with microvessels and perivascular nerves. Perivascular tissue stripping and venous distension both contribute to the reduced nitric oxide synthase in conventionally harvested veins. CONCLUSION: The new no-touch technique of saphenous vein harvesting preserves nitric oxide synthase, which suggests that improved nitric oxide availability may be an important mechanism in the success of this technique. PMID- 11854734 TI - Differential gene expression in human abdominal aorta: aneurysmal versus occlusive disease. AB - OBJECTIVE: Inflammation and atherosclerosis are present in both abdominal aortic aneurysm (AAA) and arterial occlusive disease (AOD). Changes in gene expression that underlie the development of AAA versus AOD are poorly defined. This study evaluated differences in gene expression in AAA, AOD, and control aortic tissue with human gene array technology. METHODS: RNA was isolated from human aortic specimens (seven AAA, five AOD, and five control), and complementary DNA (cDNA) probes were generated. The cDNA probes were hybridized to a human cell interaction array of 265 genes and quantitated with phosphorimaging. The data were corrected for background and were standardized to housekeeping genes. Statistical differences in individual gene expression were determined with the Kruskal-Wallis test. RESULTS: Of 265 genes studied, 11 showed statistically different expression in diseased aorta as compared with control. The following three genes were downregulated in AAA: collagen VI alpha1 (P <.037), glycoprotein IIIA (P <.006), and alpha2-macroglobulin (P <.020). The following two genes were upregulated in AOD: laminin alpha4 (P <.034) and insulin-like growth factor 2 receptor (P <.049). The following three genes were upregulated in both AAA and AOD: matrix metalloproteinase-9 (MMP-9; P <.005), intercellular adhesion molecule 1 (P <.012), and tumor necrosis factor--beta receptor (P <.022). The following three genes were downregulated in both AAA and AOD: integrin alpha5 (P <.012), ephrin A5 (P <.037), and rho/rac guanine nucleotide exchange factor (P <.028). Of 16 MMPs evaluated, only MMP-9 was significantly (P <.005) upregulated in both AAA and AOD. Evaluation results of four tissue inhibitors of metalloproteinases showed no significant difference in expression for all tissue types, although tissue inhibitor of metalloproteinase-1 trended toward upregulation in AAA (P =.053). Eight of the fifteen most highly expressed genes in all the groups were extracellular matrix or secreted proteins. Of these, only collagen VI alpha1 (P <.037) showed a significant change, although biglycan trended toward downregulation in AAA (P =.076). CONCLUSION: This study used cDNA array technology in the comparison of human control and pathologic aortic tissue. Six genes had similar differential expression in both AAA and AOD as compared with control. Even more interesting were differences between AAA and AOD in the expression of five genes. These data suggest a similarity in genetic expression for both AAA and AOD, with altered expression of several genes playing a role in disease differentiation. PMID- 11854736 TI - Impaired vasoreactivity despite an increase in plasma nitrite in patients with abdominal aortic aneurysms. AB - OBJECTIVE: This investigation was designed to determine whether differences in vasoreactivity occur in patients with abdominal aortic aneurysms (AAAs) as compared with patients with peripheral arterial occlusive disease (PAOD) or individuals (controls) without known vascular disease. METHODS: Brachial artery vasoreactivity was assessed in a blinded fashion, after endothelium-dependent (ED) and endothelium-independent (EI) flow-mediated vasodilation, in age-matched, male patients with AAAs (n = 11) or PAOD (n = 9) or in controls (n = 10). There were no significant differences in prestudy systolic or diastolic blood pressure, body mass index, or antilipidemic medications among the groups studied. Exclusion criteria included diabetes and tobacco use within 3 months. Quantitative ultrasound scan measurements of brachial artery diameters were performed at rest and after either forearm ischemia (ED) or administration of 0.4 mg sublingual nitroglycerin (EI). Plasma nitric oxide (NO(X) = NO(2) + NO(3)) was measured with the Saville assay. Asymmetric dimethylarginine, an endogenous inhibitor of NO(X) synthase, was measured with liquid chromatography. RESULTS: Initial brachial artery diameters were not significantly different among the groups studied (4.85 +/- 0.18 mm for AAA group, 4.82 +/- 0.17 mm for PAOD group, 4.68 +/- 0.20 mm for controls). ED and EI vasodilation was significantly less (P =.02 and.03, respectively) in the AAA group (-1.71 +/- 1.52 and 8.33 +/- 1.13, respectively) when compared with the controls (2.96 +/- 1.04 and 13.88 +/- 2.16, respectively). However, plasma NO(X) was significantly increased (P =.01) in the AAA group (7.86 +/- 0.85 micromol/L) as compared with both controls (5.13 +/- 0.63 micromol/L) and PAOD (4.85 +/- 0.46 micromol/L). Asymmetric dimethylarginine levels were decreased in the AAA group (0.34 +/- 0.05 micromol/L) as compared with the PAOD group (0.46 +/- 0.09 micromol/L). No correlation existed between aneurysm size and ED or EI vasodilation or plasma NO(X). CONCLUSION: This study is the first to document a divergence between ED and EI vasoreactivity and systemic NO metabolites in patients with AAAs. It is speculated that a dysfunctional vessel wall response, rather than a lack of NO, may be important in the pathogenesis of AAAs. PMID- 11854737 TI - Expression of heat shock protein-70 by dendritic cells in the arterial intima and its potential significance in atherogenesis. AB - OBJECTIVE: Overexpression of heat shock proteins (HSPs) is an important means of cell protection during physiologic stress such as occurs during atherogenesis. Immune responses are early events in atherosclerosis, with recent studies indicating that both humoral and cellular autoimmune processes in atherogenesis are directed toward HSPs. Dendritic cells are the key cells in the initiation and regulation of immune responses. This study examined whether HSP70 is overexpressed by dendritic cells in atherosclerotic lesions. METHODS: Twenty-six carotid artery and 16 aortic specimens obtained at endarterectomy and aortic reconstruction were examined with immunohistochemical techniques. The nature of cells that overexpressed HSP70 was studied in consecutive sections that were double stained with antibodies to HSP70 and cell type-specific markers, including CD1a and fascin (to identify dendritic cells), CD14 (monocytes), CD68 (macrophages), CD3 (T cells), CD15 (mast cells), von Willibrand factor (endothelial cells), and alpha-smooth muscle actin (smooth muscle cells). Staining with HLA-DR and CD1d was used to identify cells involved in antigen presentation. RESULTS: In advanced atherosclerotic lesions, several cell types, including monocytes, macrophages, dendritic cells, and smooth muscle cells, overexpressed HSP70. In contrast, in early atherosclerotic lesions, only dendritic cells overexpressed HSP70. Dendritic cells that overexpressed HSP70 frequently contacted T cells and also expressed HLA-DR. Furthermore, dendritic cells that clustered with T cells expressed CD1d, a unique molecule responsible for presenting lipid antigens. CONCLUSION: The results suggest that direct contacts between activated dendritic cells that overexpress HSP70 and T cells might be responsible for T cell activation and might facilitate the presentation of lipid antigens to T cells directly within the arterial wall. In early intimal lesions, HSP70 is overexpressed exclusively by dendritic cells, which suggests that dendritic cells might be involved in the early phases of atherogenesis. PMID- 11854738 TI - Transient advanced mental impairment: an underappreciated morbidity after aortic surgery. AB - OBJECTIVES: To determine the incidence, risk factors, and associated morbidity of transient advanced mental impairment (TAMI) after aortic surgery. METHODS: We retrospectively studied the charts of 188 consecutive patients undergoing elective aortic reconstruction during a recent 6-year period at a university hospital. All patients were lucid on admission and nonintubated at the time of evaluation at least 2 days after operation. TAMI was defined as disorientation or confusion on 2 or more postoperative days. Preoperative, intraoperative, and postoperative clinical variables were examined statistically for associations with TAMI. RESULTS: Fifty-three patients (28%) had development of TAMI 3.9 plus minus 2.8 days after operation. Stepwise logistic regression analysis selected the following independent predictors for TAMI: age >65 years (odds ratio [OR], 7.9; 95% confidence interval [CI], 2.7 to 23.7), American Society of Anesthesiology physical status classification >3 (OR, 2.8; 95% CI, 1.3 to 5.9), diabetes mellitus (OR, 3.4; 95% CI, 1.2 to 9.8), old myocardial infarction (OR, 2.4; 95% CI, 1.1 to 5.3), and hypertension (OR, 2.3; 95% CI, 1.0 to 5.3). Alcohol consumption was not significantly associated with TAMI. In the postoperative period, patients with TAMI were more likely to have hypoxia (P <.001), a need for reintubation (P <.001), pneumonia (P <.001), congestive heart failure (P =.003), and kidney failure (P =.05). In addition, patients with TAMI had a longer duration of endotracheal intubation (3.7 plus minus 7.8 vs 0.6 plus minus 1.2 days, P <.001), stay in the intensive care unit (8.9 plus minus 9 vs 3.9 plus minus 2 days, P <.001), and postoperative hospital stay (14.8 plus minus 11 vs 9.2 plus minus 5 days, P <.001) than patients without TAMI. Twenty (38%) patients with TAMI were discharged to intermediate-care facilities, compared with 11 (8%) patients without TAMI (P <.001). Postoperative variables conferring the largest relative risks for development of TAMI included oxygen saturation less than 92% (5.4), the need for reintubation (3.3), congestive heart failure (3.3), and pneumonia (3.2). TAMI, conversely, conferred the largest relative risks for development of postoperative congestive heart failure (15.3), the need for reintubation (9.3), pneumonia (7.1), and the need for ICU readmission (3.8). CONCLUSIONS: These data show that TAMI is prevalent among patients undergoing aortic reconstruction and is associated with dramatically increased morbidity and postoperative hospitalization rates. PMID- 11854739 TI - Palmar hyperhidrosis: evidence of genetic transmission. AB - BACKGROUND: Primary palmar hyperhidrosis is a condition marked by excessive perspiration and is reported to have an incidence of 1% in the Western population. It is a potentially disabling disorder that interferes with social, psychological, and professional activities. Over the past several years, several investigators have reported a positive family history in their patients treated for hyperhidrosis. To date, the cause is unknown; furthermore, epidemiologic data are scarce and inadequate. METHODS: To characterize the genetic contribution to hyperhidrosis, we conducted a prospective study of 58 consecutive patients with palmar, plantar, or axillary hyperhidrosis treated with thoracoscopic sympathectomy from September 1993 to July 1999. Forty-nine of the 58 probands volunteered family history data for these analyses (84% response rate). A standardized questionnaire was administered during the postoperative visit or by phone interview, and a detailed family history was obtained. The same questionnaire was also administered to a set of 20 control patients. The familial aggregation of hyperhidrosis has been quantified by estimating the recurrence risks to the offspring, parents, siblings, aunts, uncles, and cousins of 49 probands and 20 controls. We estimated the penetrance by use of a genetic analysis program. RESULTS: Thirty-two of 49 (65%) reported a positive family history in our hyperhidrosis group, and 0% reported a positive family history in our control group. A recurrence risk of 0.28 in the offspring of probands compared with frequency of 0.01 in the general population provides strong evidence for vertical transmission of this disorder in pedigrees and is further supported by the 0.14 risk to the parents of the probands. The results indicate that the disease allele is present in about 5% of the population and that one or two copies of the allele will result in hyperhidrosis 25% of the time, whereas the normal allele will result in hyperhidrosis less than 1% of the time. CONCLUSIONS: We conclude that primary palmar hyperhidrosis is a hereditary disorder, with variable penetrance and no proof of sex-linked transmission. However, this does not exclude other possible causes, and we anticipate that genetic confirmation of this disorder may lead to earlier diagnoses and advances in medical and psychosocial interventions. PMID- 11854740 TI - Endoluminal stent graft repair of aortobronchial fistulas. AB - OBJECTIVE: To describe our experience with endoluminal stent graft repair of aortobronchial fistulas. METHODS: We reviewed the records of patients treated with endoluminal stent grafting of aortobronchial fistulas at a private teaching hospital. All patients underwent the following diagnostic studies: computed tomography, angiography, bronchoscopy, and transesophageal echocardiography. With standard endovascular techniques, two different devices were implanted. RESULTS: Between March 1997 and October 2000, we treated four patients with postsurgical fistulas. The patients were diagnosed with hemoptysis between 3 and 23 years after aortic replacement grafting for thoracic aneurysms. Diagnostic studies varied in their ability to find the fistula. Transesophageal echocardiography most reliably demonstrated the fistula in the patients. All were successfully treated by exclusion with endoluminal stent grafting. The patients had no complications and no further episodes of hemoptysis. CONCLUSION: Endoluminal stent grafting of aortobronchial fistulas is feasible and may become the preferred method of management in patients at high risk. PMID- 11854741 TI - Color-flow duplex scanning of the leg arteries by use of a new echo-enhancing agent. AB - OBJECTIVE: This was a dose-finding and effectiveness study of a newly developed contrast-enhancing agent, sulphur hexafluoride (SF(6)), in patients with peripheral arterial disease in whom the therapeutic policy could not be established on the basis of standard color-flow duplex scanning of the leg arteries. METHODS: In this open-label, randomized, dose-ranging, crossover design, 14 patients in whom the assessment of vessel patency was difficult because of poor visibility (low-flow state) or extensive wall calcifications were studied. Contrast-enhanced duplex scanning was performed on the upper leg (n = 4), lower leg (n = 6), or pedal (n = 4) arteries after intravenous injection of four different dosages of SF(6). The results were compared with those from selective angiography of the vessel of interest. Contrast duration and agreement about the diagnosis and the confidence in the diagnosis were obtained before and after administration of the contrast agent. RESULTS: No adverse effects of the contrast agent were seen. Overall agreement was reasonable with regard to vessel patency between contrast-enhanced duplex scanning and angiography (71%). Nine of 14 vessels (64%) appeared open when contrast was applied. In four cases this could not be confirmed by angiography; in two of these cases this was due to the presence of collateral vessels. All vessels that appeared occluded with the contrast agent were also occluded on the angiogram. The confidence in the diagnosis increased from 56% to 91% after contrast administration (P <.0001). CONCLUSION: SF(6)-enhanced color-flow duplex scanning is a safe method that may improve the assessment of the patency of leg arteries, particularly in low-flow states. The visualization of collateral vessles during (enhanced) duplex scanning may be misleading because they may be regarded as the vessel of interest. PMID- 11854742 TI - Successful endografting with simultaneous visceral artery bypass grafting for severely calcified thoracoabdominal aortic aneurysm. AB - We present two cases of severely calcified thoracoabdominal aortic aneurysm treated by means of endografting with a retrograde aortomesenteric bifurcated bypass graft reconstructing the celiac axis and superior mesenteric artery. To avoid spinal ischemia, we monitored evoked spinal cord potential and performed an occlusion test of the intercostal arteries using a retrievable stentgraft. No change in evoked spinal cord potential was noted, and no endoleaks or complications, including paraplegia, were observed. This procedure is a feasible and less-invasive treatment for severely calcified thoracoabdominal aortic aneurysms. PMID- 11854743 TI - Coagulopathy as a result of factor V inhibitor after exposure to bovine topical thrombin. AB - We describe a case of severe coagulopathy after mesenteric revascularization. Laboratory investigation results revealed the presence of plasma inhibitors of factor V believed to result from exposure to bovine thrombin used for intraoperative hemostasis. Vascular and cardiothoracic surgeons commonly use topical thrombin for surgical hemostasis, and many patients undergo multiple exposure. More patients likely have factor V inhibitors develop than has previously been realized, and this may account for some otherwise unexplained postoperative coagulation disorders. This report may alert surgeons to coagulation disturbances that can result from exposure to bovine thrombin and provide guidelines for diagnosis and management. PMID- 11854744 TI - Gene expression profiling of abdominal aortic aneurysms. PMID- 11854745 TI - Regarding "Prospective randomized study of carotid endarterectomy with PTFE versus collagen impregnated Dacron (Hemashield) patching: perioperative (30-day) results". PMID- 11854746 TI - Regarding "Heparin modulates integrin function in human platelets". PMID- 11854747 TI - Regarding "The 50th anniversary of abdominal aortic reconstruction". PMID- 11854748 TI - Regarding "Preprocedural risk stratification: identifying an appropriate population for carotid stenting". PMID- 11854749 TI - Regarding "High prevalence of mild hyperhomocysteinemia in patients with abdominal aortic aneurysm". PMID- 11854750 TI - A surgeon's obligations when performing new procedures. AB - A junior member of the vascular surgery faculty has done two robotic abdominal aneurysmectomies. Although he attended a standard 2-day training course conducted by the instrument manufacturer, performed several similar procedures in the animal laboratory, and read all the recent pertinent literature, technical errors resulted in significant morbidity in his first two patients. He proposes to use the technique once again on a hospitalized patient. His troubled chairman has called you for an opinion on whether an ethical question has now arisen. There are no specific credentialling standards for robotic surgery at this institution. Where might we most expect to see an ethical problem in the surgeon's management of the new case? PMID- 11854751 TI - Polycystin-2 is an intracellular calcium release channel. AB - Polycystin-2, the product of the gene mutated in type 2 autosomal dominant polycystic kidney disease (ADPKD), is the prototypical member of a subfamily of the transient receptor potential (TRP) channel superfamily, which is expressed abundantly in the endoplasmic reticulum (ER) membrane. Here, we show by single channel studies that polycystin-2 behaves as a calcium-activated, high conductance ER channel that is permeable to divalent cations. Epithelial cells overexpressing polycystin-2 show markedly augmented intracellular calcium release signals that are lost after carboxy-terminal truncation or by the introduction of a disease-causing missense mutation. These data suggest that polycystin-2 functions as a calcium-activated intracellular calcium release channel in vivo and that polycystic kidney disease results from the loss of a regulated intracellular calcium release signalling mechanism. PMID- 11854752 TI - Arfaptin 2 regulates the aggregation of mutant huntingtin protein. AB - Huntington's disease (HD) is an inherited neurodegenerative disorder. Here we demonstrate that expression of arfaptin 2/POR1 (partner of Rac1) in cultured cells induces the formation of pericentriolar and nuclear aggregates, which morphologically resemble mutant huntingtin aggregates characteristic of HD. Endogenous arfaptin 2 localizes to aggregates induced by expression of an abnormal amino-terminal fragment of huntingtin that contains polyglutamine (polyQ) expansions. A dominant inhibitory mutant of arfaptin 2 inhibits aggregation of mutant huntingtin, but not in the presence of proteasome inhibitors. Using cell-free biochemical assays, we show that arfaptin 2 inhibits proteasome activity. Finally, we show that expression of arfaptin 2 is increased at sites of neurodegeneration and the protein localizes to huntingtin aggregates in HD transgenic mouse brains. Our data suggest that arfaptin 2 is involved in regulating huntingtin protein aggregation, possibly by impairing proteasome function. PMID- 11854753 TI - Myosin-X is an unconventional myosin that undergoes intrafilopodial motility. AB - Filopodia are thin cellular protrusions that are important in cell motility and neuronal growth cone guidance. The actin filaments that make up the core of a filopodium undergo continuous retrograde flow towards the cell body. Surface receptors or particles can couple to this retrograde flow and can also move forward to the tips of filopodia, although the molecular basis of forward transport is unknown. We report here that myosin-X (Myo10 or M10), the founding member of a novel class of myosins, localizes to the tips of filopodia and undergoes striking forward and rearward movements within filopodia, which we term intrafilopodial motility. The movements of the GFP-M10 puncta correspond to forward and rearward movements of phase-dense granules along the filopodia. Finally, overexpressing full-length M10 (but not truncated forms of M10) causes an increase in the number and length of filopodia, indicating that M10 or its cargo may function in filopodial dynamics. The localization and movements of M10 strongly suggest that it functions as a motor for intrafilopodial motility. PMID- 11854754 TI - Toward making 'IMEs' independent. PMID- 11854755 TI - Medical examiners: independent or ignorant? PMID- 11854756 TI - Toward making IMEs independent: balancing the source of work. PMID- 11854757 TI - How an audit of epidural patients in a community hospital setting resulted in the development of a formal acute pain management service. AB - PURPOSE: To describe the results of an audit of patients who received epidural analgesics postoperatively and the subsequent development of a formal acute pain management service in a community hospital. METHODS: To understand how epidural analgesia was being used to treat postoperative pain at the Peterborough Regional Health Centre, Peterborough, Ontario, a retrospective chart review was performed. Audits were performed on 178 patients who had received epidural analgesia postoperatively from October 1994 to May 1995. Data pertaining to demographics, epidural analgesia, pain scores and side effects were collected. RESULTS: Sixty one per cent of patients received bupivacaine/ fentanyl infusions, and 39% received epidural morphine boluses. More than 60% of patients reported no pain postoperatively. Patients who received bupivacaine/fentanyl were more likely than those who received epidural morphine to also receive co-analgesia and transitional analgesia. Patients who received epidural morphine were more likely than those who received bupivacaine fentanyl to experience respiratory depression, hypotension and pruritus. Patients were followed by the anesthesiologist who provided the anesthetic. Anesthesiologists practised independently, and formal policies and procedures did not exist. CONCLUSIONS: As a result of the audit, an acute pain management service was developed. This included a team that did daily rounds and consisted of a nurse clinician and an anesthesiologist who was assigned to the service on a weekly basis. A committee was created, and formalized policies and procedures were established. Standardized order sheets, data sheets and a computerized database were developed. Reports for administrative and quality improvement purposes were generated monthly. Education programs were developed. Co-analgesia and transitional analgesia are now part of routine care, and epidural catheter placement close to the site of incision is encouraged. A postoperative nausea and vomiting algorithm, and a treatment regimen for pruritus have also been implemented. PMID- 11854758 TI - Mechanisms of sucrose and non-nutritive sucking in procedural pain management in infants. AB - The administration of sucrose with and without non-nutritive sucking (NNS) has been examined for relieving procedural pain in newborn infants. The calming and pain-relieving effects of sucrose are thought to be mediated by endogenous opioid pathways activated by sweet taste. The orogustatory effects of sucrose have been demonstrated in animal newborns, and in preterm and full term human infants during painful procedures. In contrast to sucrose, the analgesic effects of NNS are hypothesized to be activated through nonopioid pathways by stimulation of orotactile and mechanoreceptor mechanisms. Although there is uncertainty as to whether the effects of sucrose and NNS are synergistic or additive, there is sufficient evidence to support the efficacy of combining the two interventions for procedural pain relief in infants. In this review article, the underlying mechanisms of sucrose and NNS, separately and in combination for relieving procedural pain in preterm and full term infants, are examined. Clinical and research implications are addressed. PMID- 11854759 TI - Children's pain: a mandate for change. AB - There have been tremendous research advances in the past 15 years in knowledge about children's pain, and strategies for recognizing and managing that pain. However, the clinical care of children in pain remains a challenge. Children's pain continues to be frequently unrecognized, dismissed or ineffectively managed. A loud call for change is being voiced by physicians, nurses, children and their families. A review of the literature was conducted to document this issue. Starting with a Medline search of the key word 'child* + pain' and continuing with a snowball technique, articles and resources addressing children's pain were collected. Resources presented or published after 1990 were particularly sought because they theoretically reflect both current knowledge about children's pain and the implementation of this knowledge in practice. Unfortunately, although information on pain is available to help children, in many instances, it is not being used. The purpose of the present paper is twofold - to present an overview of current knowledge of children's pain, and factors that hinder its effective assessment and management; and to present a mandate for change. Children's postoperative pain is highlighted in this paper as an example of the gap between pain knowledge and clinical practice. Although treatment strategies differ across different types of pain, children's conditions and ages, the principles and mandate for change discussed in this paper are directly relevant to all categories of children's pain. PMID- 11854760 TI - An analysis of articles on neonatal pain published from 1965 to 1999. AB - OBJECTIVE: To analyze articles on neonatal pain indexed in MEDLINE from 1965 to 1999. METHODS: Papers published since 1965 and indexed in Medline (PubMed ) were considered. The search strategy was: 'pain' (MeSH terms) OR 'pain' (Title word) OR 'pain' (All Fields), AND 'infant, newborn' (MeSH terms). After the search was completed, a printed copy of the references was obtained, and each reference was reviewed to establish its pertinence to the study's objective. For each article, the year of publication, the journal of publication, the language of the journal, the country of the first author, the type of article and the subject were analyzed. RESULTS: Only 545 of 2490 references identified by the MEDLINE search were considered to deal with neonatal pain. Few articles on neonatal pain were published from 1965 to 1985, but the number of articles published on this topic increased sharply after 1985. Most of the papers were published in English (84.6%). The United States, Canada and the United Kingdom were the main countries that published neonatal pain articles (38.6%, 14.2% and 13.3%, respectively). Clinical research articles (31.7%) were published most frequently; the main subjects of all the articles were pain related to colic (20.4%), general neonatal pain issues (13.6%) and procedural pain (13.4%). Pediatrics and Archives of Diseases in Childhood were the journals that published the most articles on neonatal pain (27 and 22, respectively). CONCLUSIONS: Pain in neonates was a neglected subject of publication until the mid-1980s, and, currently, only a few countries seem interested in this type of pain. This lack of interest may be related to the undertreatment of pain in the neonatal period. PMID- 11854761 TI - Multiple lessons. PMID- 11854762 TI - The McGill Pain Questionnaire, Japanese version, reconsidered: confirming the theoretical structure. AB - BACKGROUND: Based on a tripartite theoretical model of pain, the Pain Rating Index (PRI) of the McGill Pain Questionnaire (MPQ) continues to be one of the most frequently used instruments to measure clinical pain. However, language and cultural barriers have hindered its wide use and standardization in Japan. Although a number of exploratory factor analysis studies have failed to support consistently the theoretical structure of the MPQ, a few previous confirmatory factor analysis studies did statistically support the a priori model. OBJECTIVE: To test, through confirmatory factor analysis (CFA), the theoretical structure of a Japanese version of the MPQ (JMPQ), which followed a format similar to that of the original MPQ. DESIGN: This study used CFA on prospectively collected data from 199 consecutive outpatients with chronic pain at a university hospital to test the theoretical structure of the JMPQ. RESULTS AND CONCLUSIONS: CFA was completed on the first 16 PRI subclass scores; this process yielded a well fitting final model that explained 92% of the covariance in the observed data. The results supported the hypothesis that the sensory, affective and evaluative subscales of the PRI are representative of the multidimensionality of the pain experience, with minimal overlap. It is suggested that the theoretical structure of the MPQ is maintained in the JMPQ used in this study. Therefore, this study is the first step toward standardization of the JMPQ, serving as a cultural bridge in the field of pain medicine between Japan and English-speaking nations such as Canada. PMID- 11854763 TI - Effects of flotation-REST on muscle tension pain. AB - The purpose of the present study was to investigate whether the floating form of the restricted environmental stimulation technique (REST) may be applied within the field of pain relief. Flotation-REST consists of a procedure whereby an individual is immersed in a tank filled with water of an extremely high salt concentration. Thirty-seven patients (14 men and 23 women) suffering from chronic pain consisting of aching muscles in the neck and back area participated in the study. They were randomly assigned to either a control group (17 participants) or an experimental group (20 participants). The experimental group received nine opportunities to use the flotation-REST technique in the water tank over a three week period. The results indicated that the most severe perceived pain intensity was significantly reduced, whereas low perceived pain intensity was not influenced by the floating technique. Further, the results indicated that circulating levels of the noradrenaline metabolite 3-methoxy-4 hydroxyphenylethyleneglycol were reduced significantly in the experimental group but not in the control group following treatment, whereas endorphin levels were not affected by flotation. Flotation-REST treatment also elevated the participants' optimism and reduced the degree of anxiety or depression; at nighttime, patients who underwent flotation fell asleep more easily. The present findings describe possible changes, for the better, in patients presenting with chronic pain complaints. PMID- 11854764 TI - NSAID antinociception measured in a chemical and a thermal assay in mice. AB - The antinociceptive activity of several nonsteroidal anti-inflammatory drugs (NSAIDs) that were administered either intraperitoneally or intrathecally was assessed in mice by two algesiometric tests. The first was the writhing test, which assessed the abdominal constrictions that were induced by intraperitoneal acetic acid (a chemical test), and the second was the tail flick test, which measured pain responses to heat stimuli. The corresponding effective doses and their relative potencies were compared because these tests use different nociceptive stimuli with different transmission pathways. The intraperitoneal and intrathecal dose-response curves for the antinociception induced by every NSAID that was tested were parallel in the writhing test. In the tail flick test, however, only the intraperitoneal and intrathecal dose-response curves for ketoprofen, piroxicam, naproxen, nimesulide, paracetamol and diclofenac were parallel. The results obtained in this study confirm that NSAIDs possess different abilities to induce inhibition of cyclooxygenase, and they can be indirectly assessed by their different antinociceptive activities, depending on the algesiometric assays that are used. The antinociception of most NSAIDs might involve central mechanisms. The findings demonstrate the increasing importance of the spinal cord in processing and modulating nociceptive input, because intrathecal administration of NSAIDs is always more effective (in terms of potency) than systemic administration; thus, the antinociceptive efficacy of NSAIDs strongly depends on the algesiometric assay that is used and on the type of the nociceptive stimulus to which the test subject is exposed. PMID- 11854765 TI - A case of pain, factitious disorder and boundary violations. AB - Although health professionals are usually familiar with factitious disorders, evaluating such cases may be complicated, particularly in the legal arena. This article describes a patient who presented with pain complaints to numerous doctors. Eventually, a malpractice suit was brought against one of the patient's physicians who had diagnosed her condition as multiple personality disorder (dissociative identity disorder). Factitious disorder and the doctor's prescription of pain medications were issues raised during the trial. In view of the issue of harm, physicians' responsibilities and limitations during ongoing medical care are addressed in this case report. PMID- 11854766 TI - Swelling of the leg, deep venous thrombosis and the piriformis syndrome. AB - BACKGROUND: The piriformis syndrome, which was first described 60 years ago, is a well recognized cause of sciatica, leg pain and low back pain, due to the entrapment of the sciatic nerve in the piriformis and other rotator muscles. Very few complications relating to this syndrome have been described. AIMS: To discuss how the piriformis syndrome may cause venous engorgement in the lower limb, and how the piriformis syndrome should be included as a possible cause of acute deep venous thrombosis in a not initially swollen leg. Both complications can occur independently. METHODS: Two cases of swelling of the leg and acute deep venous thrombosis independently associated with the piriformis syndrome are presented. CONCLUSIONS: Swelling of the leg and deep venous thrombosis are possible complications of the piriformis syndrome that occur due to entrapment of nerves and vessels within the leg, secondary to a severe spasm and hypertrophy of the piriformis and other rotator muscles. PMID- 11854767 TI - Proceedings of the inaugural meeting of the Canadian Consortium for the Investigation of Cannabinoids. PMID- 11854768 TI - The cannabinoids: an overview. Therapeutic implications in vomiting and nausea after cancer chemotherapy, in appetite promotion, in multiple sclerosis and in neuroprotection. AB - The present paper describes the historical use of cannabis, starting with its use in Assyria and China. Recent advances in the understanding of the molecular basis of cannabis action are explained, including the identification of the cannabinoid receptors CB(1) and CB(2), as well as the isolation of endogenous cannabinoids from the brain and periphery. The use of delta(9)-tetrahydrocannabinol as an anti vomiting and anti-nausea drug for cancer chemotherapy, and as an appetite enhancing agent is described. Clinical work in multiple sclerosis, which may lead to the approval of tetrahydrocannabinol as a drug for this condition, is presented. Preclinical and clinical investigations with cannabidiol, a non psychotropic cannabis constituent, are also described. Recent work with cannabidiol in animal models of rheumatoid arthritis may lead to clinical investigations. A synthetic cannabinoid, HU-211 (Dexanabinol), is in advanced clinical stages of investigation as a neuroprotectant in head trauma. The above clinical approaches may ultimately lead to the realization that cannabinoids are valuable clinical drugs in numerous fields. PMID- 11854769 TI - Cannabinoids and pain. AB - Cannabinoids have been used to treat pain for many centuries. However, only during the past several decades have rigorous scientific methods been applied to understand the mechanisms of cannabinoid action. Cannabinoid receptors were discovered in the late 1980s and have been found to mediate the effects of cannabinoids on the nervous system. Several endocannabinoids were subsequently identified. Many studies of cannabinoid analgesia in animals during the past century showed that cannabinoids block all types of pain studied. These effects were found to be due to the suppression of spinal and thalamic nociceptive neurons, independent of any actions on the motor systems. Spinal, supraspinal and peripheral sites of cannabinoid analgesia have been identified. Endocannabinoids are released upon electrical stimulation of the periaqueductal gray, and in response to inflammation in the extremities. These observations and others thus suggest that a natural function of cannabinoid receptors and their endogenous ligands is to regulate pain sensitivity. The therapeutic potential of cannabinoids remains an important topic for future investigations, with previous work suggesting utility in clinical studies of cancer and surgical pain. New modes of delivery and/or new compounds lacking the psychotropic properties of the standard cannabinoid ligands offer promise for cannabinoid therapeutics for pain. PMID- 11854770 TI - Medicinal use of cannabis: history and current status. AB - OBJECTIVE: To provide an overview of the history and pharmacology of cannabis in relation to current scientific knowledge concerning actual and potential therapeutic uses of cannabis preparations and pure cannabinoids. METHODS: The literature on therapeutic uses of cannabis and cannabinoids was assessed with respect to type of study design, quality and variability of data, independent replications by the same or other investigators, magnitude of effects, comparison with other available treatments and reported adverse effects. The results of this review were also compared with those of major international reviews of this topic in the past five years. CONCLUSIONS: Pure tetrahydrocannabinol and several analogues have shown significant therapeutic benefits in the relief of nausea and vomiting, and stimulation of appetite in patients with wasting syndrome. Recent evidence clearly demonstrates analgesic and anti-spasticity effects that will probably prove to be clinically useful. Reduction of intraocular pressure in glaucoma and bronchodilation in asthma are not sufficiently strong, long lasting or reliable to provide a valid basis for therapeutic use. The anticonvulsant effect of cannabidiol is sufficiently promising to warrant further properly designed clinical trials. There is still a major lack of long term pharmacokinetic data and information on drug interactions. For all the present and probable future uses, pure cannabinoids, administered orally, rectally or parenterally, have been shown to be effective, and they are free of the risks of chronic inflammatory disease of the airways and upper respiratory cancer that are associated with the smoking of crude cannabis. Smoking might be justified on compassionate grounds in terminally ill patients who are already accustomed to using cannabis in this manner. Future research will probably yield new synthetic analogues with better separation of therapeutic effects from undesired psychoactivity and other side effects, and with solubility properties that may permit topical administration in the eye, or aerosol inhalation for rapid systemic effect without the risks associated with smoke inhalation. PMID- 11854771 TI - Cannabinoids and the immune system. AB - The effect of cannabimimetic agents on the function of immune cells such as T and B lymphocytes, natural killer cells and macrophages has been extensively studied over the past several decades using human and animal paradigms involving whole animal models as well as tissue culture systems. From this work, it can be concluded that these drugs have subtle yet complex effects on immune cell function and that some of the drug activity is mediated by cannabinoid receptors expressed on the various immune cell subtypes. However, the overall role of the cannabinoid system of receptors and ligands in human health and disease is still unclear and requires extensive elucidation. Further studies will define the precise structure and function of the putative immunocannabinoid system, the potential therapeutic usefulness of these drugs in chronic diseases such as acquired immune deficiency syndrome and multiple sclerosis, the effects of these agents on tumour growth and induction of apoptosis, and the potential anti inflammatory and proinflammatory properties of cannabimimetic compounds. It is likely that the cannabinoid system, along with other neuroimmune systems, has a subtle but significant role in the regulation of immunity and that this role can eventually be exploited in the management of human disease. PMID- 11854772 TI - Does experience influence judgments of pain behaviour? Evidence from relatives of pain patients and therapists. AB - OBJECTIVE: Judgments about an individual's pain can be profoundly important to sufferers. Relatively few studies have examined variables that may affect observers' judgments of the pain of others. The present article reports two studies investigating the relationship between different kinds of exposure to pain problems and observers' ratings of the pain intensity of patients. DESIGN: In the first study, 82 observers were classified into groups with positive and negative family histories of chronic pain. They viewed a videotape showing the facial expressions of shoulder pain patients undergoing physiotherapy assessments and rated the pain experienced by the subjects. In the second study, the data from observers having no experience with pain problems were compared with data collected from therapists having considerable experience with pain problems. RESULTS: Observers with a positive family history of chronic pain attributed greater pain to the patients than those with a negative family history of chronic pain. Professionals' pain judgments were lower than those of control subjects. CONCLUSIONS: Together, the findings imply that one's experiences with the different problems of pain patients may affect pain judgments. Alternative interpretations of the findings are considered. PMID- 11854774 TI - Pain and cognitive status among nursing home residents in Canada. AB - BACKGROUND: There is little information available on the rates of pain in institutionalized elderly persons, and this is particularly true for Canada. OBJECTIVES: To provide information about the prevalence and clinical correlates of pain in a sample of Canadian nursing homes, to determine whether residents with cognitive impairment experience lower rates of health conditions associated with pain (eg, arthritis) than residents without cognitive impairment and to determine whether the associations (ie, odds ratios) for pain with such health conditions vary as a function of cognitive status. DESIGN: The study is based on a secondary analysis of data collected with the minimum data set (MDS 2.0). SETTING AND PARTICIPANTS: The study comprised 3195 nursing home residents in Ontario, Manitoba and Saskatchewan. SUBJECTS AND METHODS: All residents were assessed with the MDS 2.0 by trained clinicians (usually nurses). Pain was documented if it had occurred within the seven days before the assessment. Assessors were trained to look for overt signs of discomfort, such as wincing or verbalizations. Self-report ratings were obtained when possible. RESULTS: The overall prevalence of pain in this sample was 49.7%, and 23.7% of residents experienced pain daily. Persons with and persons without cognitive impairments did not differ with respect to the prevalence of conditions likely to cause pain and the associations of pain with such health conditions. Regional differences were found, with Ontario residents having a higher frequency and intensity of pain than their counterparts in Saskatchewan and Manitoba. This may be due, at least in part, to regional differences in nursing home admission criteria. CONCLUSIONS: The findings suggest that the prevalence of identified pain is lower among nursing home residents with higher levels of cognitive impairment. These results do not support the notion that this is a function of lower prevalence rates of pain-causing conditions in nursing home residents with dementia. Furthermore, the results do not support the view that residents with cognitive impairments are less sensitive to pain. This study highlights the need for more comprehensive tools to assess pain in persons with cognitive impairments. Nonetheless, the MDS may be a useful instrument for detecting pain in such populations, because it does not rely exclusively on self-report. PMID- 11854775 TI - An examination of pain perception and cerebral event-related potentials following carbon dioxide laser stimulation in patients with Alzheimer's disease and age matched control volunteers. AB - BACKGROUND: Pain perception is known to depend on integrated cognitive processing. Alzheimer's disease affects 5% to 10% of older adults, but the impact of this disease on pain sensitivity and report has yet to be fully investigated. AIM OF INVESTIGATION: The present study examined pain threshold, the reliability of pain report and the central nervous system processing of noxious input, as indexed by cerebral event-related potentials (CERP). METHODS: Carbon dioxide laser detection and heat pain thresholds were determined on the hand dorsum of 15 healthy older adults (Mini-Mental State Examination [MMSE] score 29.9 +/- 0.3) and 15 persons with cognitive impairment (MMSE score 12.7 +/- 6.1). Using an array of 15 silver/silver chloride scalp electrodes, the CERP and subjective rating of stimulus intensity were recorded after fixed intensity, 25 W laser stimuli. RESULTS: Compared with age-matched controls, the detection threshold for just noticeable sensation was significantly increased in elderly adults suffering from Alzheimer's disease. There was no difference in pain threshold intensity between persons with cognitive impairment and controls, although the former group was less reliable in reporting detection and pain threshold sensations. The subjective rating of a 25 W stimulus was virtually identical in both groups, and the amplitude of the major CERP component (P400) was similar; however, cognitively impaired adults exhibited a significant increase in the latency of the P400 response. CONCLUSIONS: The present findings indicate that pain perception in response to an acute heat pain stimulus is not diminished in older persons with cognitive impairment. Patients with Alzheimer's disease may be slightly less reliable in threshold pain report, although the subjective rating of evoked pain and the level of poststimulus cortical activation following noxious stimulation were found to be similar to those of controls. A longer latency of the CERP may suggest slower cortical processing of nociceptive input by persons with Alzheimer's disease. PMID- 11854776 TI - Are all older adults with persistent pain created equal? Preliminary evidence for a multiaxial taxonomy. AB - BACKGROUND: Persistent pain is grossly undertreated in older adult sufferers, despite its high prevalence in this age group. Because of its multidimensional impacts, including depression, sleep disruption and physical disability, patients with persistent pain often benefit from interdisciplinary pain clinic treatment. This treatment is expensive, however, and may not be required by all patients. The Multiaxial Assessment of Pain (MAP) has demonstrated value in predicting response to treatment in younger adults with persistent pain. OBJECTIVE: To examine the feasibility of a MAP taxonomy for community-dwelling adults age 65 years or older. PARTICIPANTS AND PROCEDURES: One hundred eight subjects with persistent pain (mean age 73.8 years, SD=8.4 years) were interviewed and data collected on demographics, pain intensity, depressive symptoms, sleep disruption, pain interference with performance of basic and instrumental activities of daily living, frequency of engagement in advanced activities of daily living, cognitive function and comorbidity. A subset of these subjects underwent physical capacities testing, including maximal isometric lift strength, dynamic lifting endurance, timed chair rise and balance. RESULTS: Analyses derived three primary clusters of patients. Cluster 1 (24%) reported less intense pain, less depression and sleep disruption, and higher activity levels. Cluster 3 (30%) suffered from more pain and were more functionally disabled. Cluster 2 (46%) had characteristics of cluster 1 and cluster 3, but with some characteristics that were clearly unique. CONCLUSIONS: While these results are preliminary and require further validation, they indicate that older adults are heterogeneous in their response to persistent pain. Future studies should be performed to examine whether the MAP taxonomy is applicable to older adults regardless of medical diagnosis. Ultimately, this information may have meaning with regard to both treatment prescribing, and the design and interpretation of intervention studies. PMID- 11854777 TI - The classification of patients with chronic pain: age and sex differences. AB - OBJECTIVE: To further develop an empirically based classification system for chronic pain patients through the examination of age and sex differences, and incorporation of pain duration in the grouping algorithm. SUBJECTS: Three hundred seventy-four chronic pain patients (300 aged 13 to 59 years; 74 aged 60 to 89 years) assessed at an outpatient, multidisciplinary pain management centre. METHODS: Patients completed measures of demographic and descriptive information, pain intensity (box rating scale), perceived disability (modified Pain Disability Index) and affective distress (Symptom Checklist-90 Revised) before multidisciplinary treatment. Standardized scores from the assessment measures were entered into a series of hierarchical, multivariate cluster analyses to identify underlying patient subgroups. RESULTS: Age-based patient groupings from prior research were partially replicated. Significant differences in clinical presentations were observed across age and sex groups. Pain duration was found to make an important contribution to the patient groupings. 'Good control' (low pain, disability, distress) and variants of 'chronic pain syndrome' (elevated pain, disability, distress) groupings were identified across all analyses. Two variants of a 'stoic' profile were identified among older patients, with low levels of distress relative to pain and perceived disability. One of these profiles was associated with long pain duration and was found only among males. Several unique clinical profiles were identified for female patients. CONCLUSIONS: There are important age and sex differences in the clinical presentations of chronic pain patients. Some older patients present with unique clinical profiles that may reflect cohort differences, and/or physiological or psychological adjustment processes. There appears to be a greater number of distinct chronic pain presentations among females. Research on the classification of chronic pain patients within homogeneous diagnostic subgroups is needed. PMID- 11854778 TI - Tic douloureux. AB - Tic douloureux is an excruciatingly painful condition that primarily affects elderly people. It consists of unilateral electric shock-like facial pains triggered by non-noxious stimulation with clear-cut pain-free intervals. It should be discriminated from all other types of facial pain by the history and physical examination. Primary treatment includes anticonvulsant drugs; if these fail or side effects prevent their use, a surgical procedure is warranted. Almost every patient with tic douloureux can be relieved of his or her pain with anticonvulsant medications or surgery. Stereotactic radiosurgery, percutaneous gangliolysis and suboccipital craniectomy with microvascular decompression are the primary surgical options. The common aspects of tic douloureux and some of the rarer variations are reviewed, and treatment options are presented. PMID- 11854779 TI - Off Label Indications. PMID- 11854780 TI - Regulation of the Immune Response by Eicosanoids: Pharmacology and Clinical Effects of Prostaglandin E in Aspirin-Sensitive Syndromes. AB - Immunomodulatory effects of prostaglandin E(2) (PGE(2)) have been documented both in vitro and in vivo. Our previous studies have examined the effects of intravenously administered PGE(2) in mast-cell-mediated diseases, including aspirin-sensitive asthma and systemic mast-cell-activation syndrome. The basis for investigations of these particular diseases has been the hypothesis that the inhibition of cyclooxygenase removes one of its products, PGE(2), that provides a critical restraint on the activation of the mast cell. Based on the beneficial effects of PGE(2) found in these studies, we have extended our investigations to an evaluation of misoprostol, the orally available analog of PGE(1). Our preliminary studies with this drug are consistent with an inhibition of mast cell activation by misoprostol, an effect observed at doses higher than currently recommended for gastric protection. The findings from these initial trials have led to the development of ex vivo whole-blood assays that assess the pharmacodynamics of misoprostol's immunomodulatory actions, which support the concept of employing higher doses to obtain sustained systemic effects. To extend these results, we have undertaken double-blinded, placebo-controlled clinical investigations to examine the effects of the higher doses of misoprostol (300- 600 &mgr;g QID) given chronically in aspirin-sensitive asthma and systemic mast cell activation. Although still ongoing, our studies have confirmed by a variety of clinical evaluations that the higher doses of misoprostol can be tolerated by many patients and appear to be safe. Based on our findings and those of others, further investigation of the therapeutic usefulness of this drug or other PGE analogs in allergic and immunologic diseases appears warranted. PMID- 11854781 TI - Misoprostol Inhibits the Cutaneous Late-Phase Allergic Response to Antigens. Results of a Double-Blind Placebo-Controlled Randomized Study and an Investigation into the Mechanism of Action. AB - We investigated the effect of misoprostol on allergen-induced cutaneous immediate and late-phase allergic reactions in a double-blind placebo-controlled randomized study. Sixteen dust-mite-allergic patients received misoprostol (200 &mgr;g) or placebo and then had skin testing on two different days. The immediate and late-phase skin response was monitored for 6 h. Skin biopsy was obtained from five selected donors at 5 h. In vitro studies included the effect of misoprostol on eosinophil chemotaxis, eosinophil survival, basophil histamine release, and cytokine production by lymphocytes. All subjects developed an immediate wheal reaction and a late-phase induration in response to dust-mite allergens after taking placebo. Misoprosol selectively inhibited the late-phase but not the immediate-phase response (p < 0.05). Histologic studies revealed a trend toward a reduced number of inflammatory cells in the skin dermis after misoprostol treatment. We also investigated the mechanism of action of misoprostol in vitro. Misoprostol significantly (p < 0.05) inhibited eosinophil chemotaxis and the production of granulocyte/macrophage colony-stimulating factor by lymphocytes at concentrations greater-than-or-equal10(minus sign8) M. However, at significantly lower concentrations (greater-than-or-equal10(minus sign12)M), misoprostol blocked cytokine-stimulated eosinophil survival. Thus, misoprostol has potent antiallergic effects and blocks the cutaneous late-phase allergic inflammation. PMID- 11854782 TI - Initial Studies on the Effect of Inhaled Misoprostol in a Guinea Pig Model of Allergic Bronchoconstriction. AB - Inhaled E series prostaglandins (PGEs) have been shown to modulate responses to both allergic and nonallergic provocation. Little is known about the effect of inhaled misoprostol on the airway and whether its antiasthmatic activity would be similar to other PGEs. In the present study, nebulized solutions of misoprostol and PGE(2) (0.3--300 &mgr;g ml(minus sign1)) effectively blocked the acute bronchospasm due to inhaled antigen challenge in actively sensitized guinea pigs. A 300-&mgr;g ml(minus sign1) solution nebulized for 10 s (about 0.25 ml), 5 min prior to challenge, provided nearly complete inhibition with significant reductions seen at 30 and 3 &mgr;g ml(minus sign1) in certain experiments. Misoprostol treatment resulted in a significant reduction in the number of eosinophils present in bronchoalveolar lavage 24 h after antigen challenge. This combination of effects suggests that inhaled misoprostol may be effective in the treatment of the acute and chronic symptoms of asthma. PMID- 11854783 TI - The Role of Mast Cells and Their Mediators in IgG-Antigen Complex-Mediated Inflammation. AB - The strategic localization of mast cells near blood vessels led us to investigate the involvement of these cells in IgG-antigen complex-mediated inflammation, using mast-cell-deficient mice and their congenic controls. Mast cells were extensively degranulated and contributed to neutrophil influx, plasma exudation, fibrin deposition, edema formation, and tissue damage. Leukotrienes and the tumor necrosis factor (TNF) from mast cells participated in neutrophil elicitation, and histamine and leukotrienes in plasma exudation, fibrin deposition, and edema formation. Reconstituting the deficient mice with mast cells restored the responses, confirming the role of mast cells and their mediators. Studies with decomplemented and C5-deficient mice indicate that mast cells were stimulated by complement early in the reaction and later by an unknown mechanism. The findings show that mast cells and their mediators, such as leukotrienes, histamine, and TNF, play an important role in the initiation of IgG-antigen complex-mediated inflammation. PMID- 11854784 TI - Effect of Misoprostol on Histamine Secretion from Isolated Rat and Human Mast Cells and Basophils. AB - Misoprostol was found to be an extremely potent inhibitor of immunologically induced histamine release from mast cells isolated from both the human colonic mucosa and submucosa/muscle. The dose-inhibition curves were bell shaped, with IC(50) values in the range 0.1--1 nM. The compound was relatively selective for these cell types. It was less active against rat peritoneal mast cells, which again exhibited a bell-shaped dose-inhibition curve, but with a maximal inhibition of only about 30%, and human basophil leucocytes, where the inhibition was progressive with an IC(50) of similar1 &mgr;M. Very high concentrations of misoprostol (greater-than-or-equal5 &mgr;M) induced histamine release from all of the cell types examined. Insofar as the colonic mast cells may represent the two human mast cell phenotypes so far identified (MC(T) and MC(TC)), these data suggest that the drug may have therapeutic applications in human allergic and inflammatory diseases in which these cells are involved. PMID- 11854785 TI - Suppression of IL-4 Production in Murine Lymphocytes by Orally Effective Prostaglandin E(1) Analogs. AB - Prostaglandins (PGs) regulate a wide variety of immunologic processes. We studied the activity of PGE(1), and two orally effective PGE(1) methyl ester analogs, misoprostol (MP) and enisoprost (EP), as inhibitors of interleukin-4 (IL-4) production in stimulated murine splenic T cells in vitro. Effective suppresion of IL-4 production was seen with all three agents with KI(50)s of 65 nM for MP and 20 nM for EP. IL-4 is primarily a product of type 2 T helper cells (Th2 cells) and plays an important role in regulating the biosynthesis of immunoglobulins, especially IgE. These in vitro studies, taken together with the recent data of others on modulation of IgE responses by EP and MP in immunized mice in vivo, suggest a possible role for these PGE(1) analogs in the treatment of IgE-mediated diseases in humans. PMID- 11854786 TI - Differential Regulation of Fcgamma Receptor Isoforms: Implications for Therapeutic Intervention. AB - Immune complex initiation of inflammatory diseases is mediated, to a large extent, by Fcgamma receptors. Recent data have underscored three essential points: (1) there is substantial structural diversity within three distinct families of Fcgamma receptors; (2) there are functionally significant alleles of different receptor isoforms; and (3) each cell type has a unique distribution of Fcgamma receptor isoforms. Structurally different Fcgamma receptors utilize distinct signal transduction pathways. Thus, in clinical immune complex disease, consideration of the predominant cell type and receptor isoform involved, as well as the genotype of the individual patient, will be important in developing and applying targeted downregulatory agents such as misoprostol. PMID- 11854787 TI - Prostaglandin E(2)-Mediated Inhibition of the Transendothelial Migration of Human T Lymphocytes. PMID- 11854788 TI - Bacteria--Mast Cell Interactions in Inflammatory Disease. AB - Chronic inflammatory diseases of the gastrointestinal tract such as ulcerative colitis and Crohn's disease are characterized by mast cell proliferation and secretion of inflammatory mediators. The determinant(s) responsible for stimulating mast cells in the intestinal mucosa is not known. We investigated the interaction of mast cells with type 1 fimbriated Escherichia coli, an opportunistic pathogen and a constituent of the normal indigenous microflora of the gut. Unlike a mutant derivative deficient in the FimH subunit of the fimbriae or nonfimbriated E. coli, type 1 fimbriated E. coli adhered avidly to mast cells. As a consequence of this interaction, the mast cells phagocytozed and killed adherent bacteria. The mast cell bactericidal activity involved generation of superoxide anion and acidification of phagocytic vacuoles. In addition, many of the mast cells had degranulated and released inflammatory mediators such as histamine. These observations have implications both for normal host defense and for the initiation and perpetuation of inappropriate inflammatory responses in the gastrointestinal tract. PMID- 11854789 TI - Acute and Chronic Effects of Misoprostol in the Control of Asthma: A Pilot Study in Five Subjects. AB - Asthma is an inflammatory disease involving the airways. E-series prostaglandins have potent anti-inflammatory effects on neutrophils and T lymphocytes. Accordingly, we tested the acute (i.e., 6 h) and chronic (i.e., 1 week) effects of oral misoprostol 200 &mgr;g in five stable asthmatics. A double-blind, randomized, crossover design was used. Airway function was assessed from measurements of forced expiratory volume in 1 s (FEV(1)). The number of puffs of the inhaled beta-adrenergic agonist bronchodilator, albuterol, taken for symptomatic rescue, was assessed by diary. FEV(1) measured serially for 6 h after 200 &mgr;g misoprostol was similar on misoprostol and placebo days in all subjects. In contrast, three of the five subjects showed appreciable increases in FEV(1) during a week of misoprostol compared to either baseline or placebo week. Two subjects showed no change in the FEV(1) with 1 week of misoprostol. All three of the subjects who kept a diary decreased the number of puffs of albuterol during the misoprostol compared to placebo period. Of note, in these three subjects, airway function improved (n = 1) or remained the same (n = 2) despite a diminution in the number of puffs of a beta-agonist administered. None of the subjects administered the 200-&mgr;g oral dose QID reported any significant symptoms or adverse reaction. In conclusion, several inferences can be drawn from this early preliminary study. These data suggest that a trial involving a large number of subjects, a longer period of treatment, and a more objective method of assessing inhaler use may be useful in evaluating the chronic effects of misoprostol in asthma. PMID- 11854790 TI - Treatment of Spasticity in Spinal Cord Injury with Dronabinol, a Tetrahydrocannabinol Derivative. AB - Spinal-cord-injured patients and the medical literature have increasingly reported anecdotes regarding tetrahydrocannabinol (THC)-induced spasmolysis. These reports motivated this trial of dronabinol, a THC derivative, for the treatment of spasticity in the spinal-cord-injured population. Five made quadriplegic patients were given oral dronabinol in escalating doses from 5 mg BID to 20 mg TID in addition to their current, but ineffective, spasmolytic regime. The pendulum drop test was used to quantify spasticity (stiffness) in the knees. The Weschler Memory Scale (WMS), Profile of Mood States (POMS), and personal interviews were administered by the clinical psychologist to evaluate any changes in the subjects' cognition and/or emotional states. Spasticity was markedly improved in two of the five subjects, unchanged in a third, fluctuated in a fourth and made progressively worse in a fifth. The WMS revealed improvement in memory skills of two subjects and no change in the other. Psychological interviews and the POMS indicated decreased vigor in all subject, but otherwise demonstrated highly individualized emotional changes as indicated by increases and/or decreases in the dysphoric mood scales. PMID- 11854791 TI - A Comparison of the Cough Profile of Fosinopril and Enalapril in Hypertensive Patients with a History of ACE Inhibitor-Associated Cough. AB - OBJECTIVE: Cough frequency and severity with fosinopril and enalapril were assessed in hypertensive patients with previous angiotensin-converting enzyme inhibitor (ACEI)-associated cough. DESIGN: Prospective, multicenter, randomized, 8-week double-blind treatment. PATIENTS: One hundred seventy-nine patients (mild to-moderate hypertension, nonsmokers, mean age 58 years; 55% females; 72% Caucasian, 6% black, 19% Hispanic) were studied. Patients with other cough etiologies, significant co-morbidity, or confounding medications were excluded. INTERVENTIONS: Patients were randomized to fosinopril 10 mg (n = 85) or enalapril 5 mg (n = 94) once daily. Dosage could be doubled for blood pressure control after 4 weeks. Outcome measurements: The primary end point was all-cough frequency based on patient daily diary ratings; a cumulative cough frequency score was calculated. Secondary end points included cough severity, nonproductive cough frequency, night awakenings, cough time of day, and spontaneously reported cough. RESULTS: Fosinopril and enalapril demonstrated similar blood pressure control. Significant cough profile differences were observed in favor of fosinopril: all-cough frequency was 40.6 plus minus 3.8 (mean plus minus SE) versus 52.8 plus minus 3.6 (p = 0.02); nonproductive cough frequency was 26.7 plus minus 3.5 versus 40.3 plus minus 3.4 (p less-than-or-equal 0.01); and cough time of day was 49.2 plus minus 5.2 versus 66.0 plus minus 5.0 (p = 0.02), for fosinopril and enalapril, respectively. Subgroup analysis revealed all-cough frequency was 33.5 plus minus 6.3 versus 56.6 plus minus 5.3 (p = 0.006) for fosinopril and enalapril, respectively, in patients who previously had cough on one of these two ACEI (predominantly enalapril). Ten (12%) fosinopril and 25 (27%) enelapril patients spontaneously reported cough (p = 0.01). CONCLUSIONS: Hypertensive patients with previous ACEI-associated cough reported less frequent cough with fosinopril compared to enalapril, based on cumulative patient diary scores and spontaneously reported cough. This difference was most apparent in the subgroup of patients who previously experienced cough associated with enalapril therapy. Patients with prior ACEI-associated cough may experience less frequent with fosinopril. PMID- 11854792 TI - Absorption and Malabsorption of Levothyroxine Sodium. PMID- 11854794 TI - The Information Age and Therapeutics. PMID- 11854793 TI - Alternative Agents in Pharmacological Management of Sickle Cell Pain Crisis Complicated by Acute Pancreatitis. AB - Pain is a prominent presenting complaint in the vaso-occlusive crisis of sickle cell anemia and that of acute pancreatitis. The treatment of pain may extend for hours or days up to weeks for an episode. Treatment by pharmacotherapy often includes opiate/opioid analgesics, nonopiate analgesics and coanalgesics. Repeated acute painful episodes expose the patient to opiate induced effects. This case report describes a treatment option that was developed in order to provide effective analgesia while minimizing the iatrogenic effects produced by commonly used opioids. The use of partial agonists, mixed agonists, nonsteroidal anti-inflammatory agents and coanalgesics can provide objective and subjective analgesia without precipitating drug seeking behaviors or significant iatrogenic effects. Pain management in this clinical setting requires a balanced biopsychosocial approach. PMID- 11854795 TI - Modulation of Renal Immune Injury by Eicosanoids. PMID- 11854796 TI - Effects of Misoprostol on Contrast-Induced Renal Dysfunction. AB - Radiographic contrast-induced nephropathy (RCIN), defined by a variable rise in serum creatinine, occurs in up to 40% of contrast radiologic procedures. A prospective, randomized double-blind study was done to determine whether misoprostol prevented or modified RCIN. Patients with a serum creatinine less than-or-equal2.0 mg dl(minus sign1), who were scheduled to undergo a radiologic contrast procedure (N = 125), were randomized to receive placebo (N = 62) or misoprostol (N = 63) given at 200 &mgr;g Q.I.D. for 72 h prior to contrast and for 48 h after contrast. Contrast significantly decreased creatinine clearance, and misoprostol significantly diminished the dysfunction. The effect was more pronounced in patients with diabetes mellitus (N = 24) and patients on nonsteroidal anti-inflammatory drugs (NSAIDs) (N = 47). Our findings are consistent with a functional role of prostaglandins in the renal response to contrast. We conclude that short-term administration of misoprostol is a useful adjunct for contrast procedures, especially in patients with diabetes and patients on NSAIDs. PMID- 11854797 TI - Sucrose Permeability: A Novel Means of Detecting Gastroduodenal Damage Noninvasively. AB - Asymptomatic damage to the upper gastrointestinal tract is a common problem that may present with life-threatening sequelae such as bleeding. This scenario is especially prevalent in the population that ingest nonsteroidal anti-inflammatory agents (NSAIDs). Currently, there exists no means to screen these patients for the presence or absence of gastroduodenal damage prior to clinical presentation. Endoscopy, which remains the "gold standard" for the detection of upper gastrointestinal damage, is a time consuming technique that requires special expertise and may not be generally available. As such, it is an inappropriate technique for the widespread screening of large populations. It is now recognized that intestinal damage can be detected by determining that intestinal permeability is increased in diseases in that affect the small intestine, such as celiac or Crohn's disease. These methods are relatively simple: The patient ingests nondigestible sugar probes that cross damage mucosa and can be detected in the urine in increased amounts. Over the last several years, we have adapted these concepts for the simple noninvasive detection of gastroduodenal damage and have demonstrated that sucrose is an interesting probe molecule that specifically reports damage from the extreme proximal end of the gastrointestinal tract. In this article we review the data that support this contention and demonstrate the clinical usefulness of this approach for the detection of gastric damage in man. PMID- 11854798 TI - Protection from Radiation-Induced Oral Mucositis by Misoprostol, a Prostaglandin E(1) Analog: A Placebo-Controlled, Double-Blind Clinical Trial. AB - Misoprostol, a prostaglandin E(1) analog, is an effective radioprotector in animal studies. Based on this evidence, a prospective, randomized, placebo controlled, double-blind study was conducted to determine if misoprostol protected the oral and pharyngeal mucosa of irradiated head and neck cancer patients from radiation mucositis. Postsurgical patients who had no detectable cancer and who were referred for postoperative irradiation were candidates for this study. Thirty-four Hines VA and 35 Loyola University patients were accrued (69 total) over a 2-year period. A misoprostol tablet (200 &mgr;g) or an identical placebo tablet was dissolved in water and administered as an oral rinse daily about 20 min before irradiation. Conventional fractionated radiotherapy, consisting of five weekly doses of 2 Gy day(minus sign1), was delivered. The degree of mucositis was scored on a scale from 0 (no mucositis) to 4. In the 17 patients randomly assigned to the misoprostol arm at the Hines VA, no advantage was seen compared to the 17 placebo-treated patients. However, there was significantly less mucositis (p < 0.01, analysis of variance) from weeks 3--6 in the 17 patients treated with misoprostol at Loyola compared to the 18 placebo treated patients. Several problems in the study were identified at the VA, including adherence to the protocol design. Other problems such as adequate mucositis scoring, radiation scatter from fillings, and, in particular, adequate timing between misoprostol and irradiation were identified at both locations. Absorption studies in healthy volunteers showed significant plasma levels at 10 min after an oral rinse, suggesting that initial clinical trials should be confined to topical misoprostol until more is known regarding the effect of misoprostol on tumors. The results of this pilot study suggest that misoprotol may protect the oral and pharyngeal mucosa from radiation-induced mocositis if adequate time between topical administration and radiation is allowed. PMID- 11854799 TI - Effects of Misoprostol on Nonsteroidal Anti-Inflammatory Drug-Induced Renal Insufficiency in Patients with Stable Chronic Renal Failure: A Double-Blind, Crossover Study. AB - Nonsteroidal anti-inflammatory drugs (NSAIDs) have the ability to induce acute deterioration in renal function by inhibiting the production of vasodilatory prostaglandins, thereby disrupting intrarenal hemodynamic balance in patients with stable, mild, chronic renal failure. Misoprostol is a synthetic prostaglandin E-1 analog suitable for oral administration and is widely used in conjunction with NSAIDs to prevent another NSAID-induced complication, gastric ulcers. This investigation utilized a prospective double-blind, randomized, placebo-controlled crossover study designed to evaluate the possible renal protective effects of misoprostol on the renal function of patients with mild, stable, chronic renal failure, who intercurrently ingested ibuprofen 800 mg 8H or indomethacin 25 mg 8H. The mean baseline glomerular filtration rate of the patients who participated in the study was 53 ml/min (misoprostol)/55 ml/min (placebo), and 57 ml/min (misoprostol)/57 ml/min (placebo) in Study I (ibuprofen) and Study II (indomethacin), respectively. At this level of renal functional impairment, the use of the selected NSAIDs did not produce further significant impairment of renal function; hence, a renal protective role for misoprostol could not be demonstrated. To solve the question posed in our study, it will be necessary to conduct future investigations utilizing a recruitment GFR and serum creatinine values in the range of 35 ml/min and less and 2.0 mg/dl and greater, respectively. PMID- 11854800 TI - The Influence of Misoprostol on the Adverse Renal Effects and Stereospecific Pharmacokinetics of Ibuprofen in Chronic Renal Insufficiency. AB - The use of nonsteroidal anti-inflammatory drugs in patients with chronic renal insufficiency (CRI) may be complicated by renal functional abnormalities due to the inhibition of renal prostaglandins. We tested the hypothesis that administration of the oral PGE1 analog, misoprostol, could attenuate the adverse renal effects of ibuprofen in patients with CRI. Because the metabolism of misoprostol and the stereoinversion of R- to S-ibuprofen involve the same metabolic pathway, the stereospecific pharmacokinetics of ibuprofen were also evaluated. In a randomized, crossover trial of six stable CRI patients (Clcr 25- 67 ml min(minus sign1)), in sodium balance on a 150 mEq Na(+) per day metabolic diet, we compared the effects of ibuprofen 600 mg qid with and without misoprostol 200 &mgr;g qid upon Clcr, Clinulin, Clpah, Na(+), and K(+) excretion during 4-h clearance studies. We also assessed stereospecific ibuprofen kinetics following single dose (acute) and after 7 days on drug(s) (chronic). Daily weights, supine blood pressures, electrolytes, osmolality, BUN, creatinine and 24 h urine collections for Clcr and Na(+) and K(+) excretions were obtained during chronic dosing. Supine and upright plasma renin activities were obtained prior to dosing and during chronic dosing for both treatment limbs. Ibuprofen alone resulted in an approximately 20% transient reduction in GFR, occurring 2--2.5 h following dosing in both the acute and chronic clearance studies. This was not affected by misoprostol. There was a greater degree of stimulation of PRA with the upright posture with misoprostol plus ibuprofen than with ibuprofen alone. There was a significant weight gain in both study limbs, but no effect of misoprostol (1.2 plus minus 0.2 kg ibuprofen alone and 1.0 plus minus 0.2 kg ibuprofen plus misoprostol, p = 0.13). Otherwise no clinically significant alteration in renal function occurred in either treatment limb. The presence of misoprostol did not alter the stereospecific pharmacokinetics of ibuprofen. We conclude that misoprostol does not significantly alter the renal effects of ibuprofen in patients with mild to moderate CRI. PMID- 11854801 TI - Misoprostol in Organ Transplantation: Effect on Rejection-Associated Growth Factor Profile, Renal Hemodynamics, and Cyclosporine Cardiotoxicity. PMID- 11854802 TI - Misoprostol Partially Inhibits the Renal Scarring of Chronic Cyclosporine Nephrotoxicity. AB - Chronic cyclosporine (CY) nephrotoxicity is a well-known complication of this immunosuppressive agent, which may, in part, be attributed to abnormalities in renal prostaglandin content. We utilized misoprostol (M), a prostaglandin E(1) analog, in a rat model of chronic CY toxicity at 7 and 28 days to determine effectiveness in prevention of the renal damage. After 7 days, there were no differences in weight change, creatinine clearance, or renal scarring in rats treated with vehicle (V), CY, or CY + M; however, renal procollagen alpha 1(I) mRNA levels were increased in CY versus V rats (p < 0.05). In contrast, after 28 days CY rats had significant reductions in weight gain, glomerular filtration rate, and renal blood flow with increases in renal scarring and procollagen alpha 1 (IV) mRNA levels (all p < 0.05 versus V). Addition of M resulted in partial but significant improvement in GFR, RBF, and procollagen alpha 1(IV) mRNA levels, with lessening of the renal scarring. Skin fibroblasts also were incubated with CY and M to assess impact on procollagen MRNA levels. CY augmented fibroblast procollagen mRNA levels which enhanced by a large dose of M, but inhibited with a moderate dose. These data suggest that M improves renal scarring induced by CY by hemodynamic and/or direct effects. PMID- 11854803 TI - A Study of the Efficacy and Safety of Moexipril in Mild to Moderate Hypertension. AB - This placebo-controlled, double-blind, multicenter study examined the efficacy, safety, and tolerability of the angiotensin-converting enzyme inhibitor, moexipril, in lowering blood pressure in mildly to moderately hypertensive patients. Patients were initially randomized into four groups, two of which received moexipril 7.5 mg per day and two received moexipril 15 mg per day, for first 12 weeks of treatment. Patients then entered a withdrawal phase with one of the groups in each dose category continuing that dose of moexipril and one receiving placebo for 12 more weeks. From 223 patients randomized initially, 190 completed the 12-week withdrawal phase. In the two dosage groups from baseline to 12 weeks, sitting diastolic blood pressure decreased from 101.1 to 92.8 mm Hg for the 7.5-mg group and from 100.7 to 91.3 mm Hg for the 15-mg group (p < 0.05 baseline to week 12 in both groups) with a significant difference between those groups attained at week 12 only (p = 0.03). By the end of the withdrawal phase (24-week evaluation), the group that continued to receive 7.5 mg moexipril decreased diastolic blood pressure by 8.2 mm Hg, whereas the corresponding placebo group decreased diastolic blood pressure by 3.7 mm Hg. Although the difference between these two groups was not significant at the 24-week end point, all other time points differed significantly between groups at p less-than-or equal 0.017. Similarly, whereas the corresponding placebo group had a mean reduction in diastolic blood pressure of 4.6 mm Hg, the group that continued 15 mg of moexipril showed a mean diastolic blood pressure reduction of 10.6 mm Hg (p < 0.001 between groups). No comparison between the two moexipril dosage groups was significant, however, during the withdrawal phase. These results during medication withdrawal indicate that moexipril is effective in significantly lowering diastolic blood pressure. PMID- 11854804 TI - Cardiovascular Monitoring in Normal Healthy Adults: A Literature Review and Recommendations for the Reporting of Disturbances of Cardiac Rhythm. AB - Continuous monitoring of the electrocardiogram has become a commonly used technique in the safety assessment of new drugs when these are administered to human beings for the first time. Disturbances of cardiac rhythm have been observed with cardiac monitoring during clinical pharmacology studies in normal healthy volunteers. It is often difficult to assess the clinical significance of these rhythms especially when these rhythms occur following the administration of new drugs to human beings for the first time. Certain arrhythmias are frequently noted in normal healthy adults, including sinus bradycardia, sinus arrhythmia, the Wenckebach type of second-degree AV block (Mobitz I), atrioventricular junctional rhythm, PAC's, and PVC's. PAC's and PVC's are commonly observed, although greater than 50 PVC's/24 hr and greater than 100 PAC's/hr are relatively rare. Atrial couplets/atrial tachycardia and ventricular couplets/nonsustained ventricular tachycardia are relatively rare, but nevertheless do occur on occassion in normal healthy adults. With these consideration in mind, a guideline for the interpretation and reporting of cardiac arrhythmias occurring during clinical pharmacology studies in normal healthy adults is presented. The absence of symptoms or structural heart disease are important considerations in the evaluation of arrhythmias in normal healthy adults. The severity and seriousness of the adverse experience should be left up to the investigator's discretion. Rechallenge with active drug or placebo should be considered on a case-by-case basis. PMID- 11854805 TI - Clinical Therapeutic Conference: Thyroid Hormone Replacement in Hypothyroidism. PMID- 11854806 TI - The Promise of Chirality. PMID- 11854807 TI - Preliminary Studies of in vitro and in vivo Effects of Misoprostol on Th-1 and Th 2 Cytokine Production. AB - Prostaglandins of the E series are known to suppress in vitro production of Th-1 cytokines such as interleukin-2 (IL-2) and interferon-gamma but have not been shown to suppress production of Th-2 cytokines such as IL-4 or IL-10. The present study used two new synthetic prostaglandin E(1) (PGE(1)) analogs with oral bioavailability, misoprostol (MP), and enisoprost (EP), to determine if these agents (1) exert suppressive effects in vitro on cytokine production by fresh unseparated mouse splenocytes and (2) are beneficial in vivo when used in conditions mediated by excessive Th-1 or Th-2 cytokine production. Preliminary in vitro studies demonstrated that both MP and EP can inhibit mitogen-stimulated Th 1 and Th-2 cytokine production in a dose-dependent fashion. Interestingly, at low doses, a stimulatory effect on interferon-gamma production was seen for both agents. In vivo studies tested the ability of parenteral administration of MP to alter outcome in the parent-into-F1 model of acute or chronic graft-vs-host disease (GVHD), entities thought to be mediated by excessive Th-1 or Th-2 cytokine production, respectively. Administration of MP to mice undergoing acute GVHD resulted in little detectable effect. However, in three independent experiments, MP administration in chronic GVHD mice consistently blocked GVHD associated lymphoproliferation. In two of three experiments, GVHD-associated autoantibody production was significantly reduced. Variability between individual mice and between experiments suggests that dosing regimens and MP preparation are of critical importance. Nevertheless, these findings raise the possibility that MP may be of benefit in the treatment of human diseases characterized by excessive Th-2 cytokine production and humoral autoimmunity, for example, human lupus. PMID- 11854808 TI - Cyclic AMP-Independent Mechanisms of Nuclear Signal Transduction by PGE(2). AB - Arachidonic acid-derived mediators induce transcription of several immediate early genes, but the molecular mechanisms underlying these responses remain poorly characterized. We designed experiments to explore the mechanisms by which PGE(2) induces expression of transcription factor c-fos in glomerular mesangial cells. Binding of PGE(2) to prostaglandin receptors in mesangial cells stimulates both adenylate cyclase and phospholipase C-linked signaling pathways. Prostaglandin E(2) (PGE(2)) induced marked and transient accumulation of c-fos mRNA, but induction of the c-fos gene occurred independent of PGE(2)-stimulated adenylate cyclase activity. These results contrast with previous experiments in NIH 3T3 cells in which PGE(2) stimulated c-fos accumulation by a cAMP-dependent mechanism. We further showed that PGE(2) induces c-fos gene expression by increasing the transactivating capacity of the serum response element. Collectively, these results provide evidence of a cAMP-independent pathway linking PGE(2) receptors to transcriptional activation in the nucleus. Thus, activation of PGE(2) receptors in different cell types leads to both cAMP independent and -dependent pathways for gene expression. PMID- 11854809 TI - Effect of Misoprostol on Mesangial Cell Growth and Collagen Metabolism. AB - Independent of etiology, progressive chronic renal failure is characterized by accumulation of mesangial matrix and collagenous materials leading to glomerular sclerosis and closure. Pharmacologic intervention aimed at ameliorating this process has significant potential for substantial clinical benefit. We, therefore, assessed the effect of misoprostol on glomerular mesangial cell growth and collagen metabolism. Studies were carried out using a rat glomerular mesangial cell line cloned in our laboratory. At the concentration tested (1 &mgr;M), neither misoprostol nor prostaglandin E(2) had any effect on glomerular mesangial cell proliferation. Misoprostol did not change the absolute synthesis rates for collagen or total protein when measured by (14)C-proline incorporation into protein-associated hydroxyproline and proline respectively. However, the amount of collagen extruded into the medium, as a percentage of protein synthesis, was decreased by 10% in misoprostol-treated cells (p = 0.042). In addition, collagen breakdown was 26% greater in misoprostol-treated cultures (p = 0.044). Misprosotol had no such effects on cell cultures subjected to mechanical stress applied as continuous stretch-relaxation cycles. These results indicate that misoprostol influences mesangial cell collagen metabolism by increasing the rate of endogenous breakdown and decreasing collagen export outside the cell. Misoprostol has no effect on mesangial cell proliferation. PMID- 11854810 TI - Misoprostol and Prednisone Treatment of Lupus Nephritis. AB - Fourteen patients were enrolled in a placebo-controlled double-blind randomized trial of 8 weeks of treatment for active lupus nephritis. Seven patients received prednisone at a dose of 1 mg kg(minus sign1) day(minus sign1) plus misoprostol at a dose of 200 &mgr;g P.O. Q.I.D.; 7 patients received prednisone plus placebo. The patients included 12 females, 2 males; 3 African-Americans, 3 Asians, 5 Hispanics, and 3 Caucasians. There were no serious side effects associated with prednisone or misoprostol treatment during the 8-week study period. Laboratory measures obtained at baseline, 4, 8, and 12 weeks included complete blood count (CBC), ESR, C reactive protein (CRP), serum creatinine, creatinine clearance, 24 h urine protein excretion, C3, C4, and anti-double stranded DNA (anti-dsDNA). Statistical analysis was conducted assessing change in measures over time in the entire study group by paired t-tests. The effect of treatment on change over time was examined by analysis of covariance. Log transformation of the variables was performed prior to statistical analysis. For the entire study group, the mean levels of ESR, CRP, 24-h protein excretion, C3, C4, and anti-dsDNA were improved at 4, 8, and 12 weeks. The mean ESR at baseline was 70 plus minus 8 compared to 42 plus minus 8 at 12 weeks (p < 0.01). The mean CRP at baseline was 0.6 plus minus 0.2 compared to 0.2 plus minus 0.1 at 12 weeks (p < 0.01). The 24-h protein excretion was 4367 plus minus 769 mg at baseline compared to 2512 plus minus 709 mg at 12 weeks (p = 0.02). The mean C3 at baseline was 40 plus minus 4 mg dl(minus sign1) compared to 60 plus minus 4 mg dl(minus sign1) at 12 weeks (p < 0.01). The mean C4 at baseline was 14 plus minus 1 mg dl(minus sign1) compared to 23 plus minus 2 mg dl(minus sign1) at 12 weeks (p < 0.01). The mean anti-dsDNA at baseline was 4268 plus minus 1780 compared to 316 plus minus 111 at 12 weeks (p < 0.001). The baseline serum creatinine (1.12 plus minus.15 mg dl(minus sign1)) and creatinine clearance (82 plus minus 15 ml min(minus sign1)) were not significantly changed at 12 weeks (1.10 plus minus 0.2 mg dl(minus sign1) and 80 plus minus 17 ml min(minus sign1), respectively). Comparing the misoprostol treatment group to the placebo group, there were no statistically significant differences for ESR, CRP, creatinine, creatinine clearance, 24-h protein excretion, C3, C4, or anti-dsDNA at any time points. However, comparing the misoprostol treatment group at 4 weeks to the placebo group, a more rapid decrease in anti-dsDNA (reduction of 3297 plus minus 2374) was observed in the misoprostol treatment group versus 304 plus minus 409 in the placebo group), as well as lower mean anti-dsDNA levels (464 plus minus 140) in the misoprostol treatment group versus 4118 plus minus 3834 in the placebo group). Also, the C3 and C4 levels at 12 weeks in the misoprostol treatment group (67 plus minus 5 and 27 plus minus 3 mg dl(minus sign1), respectively) were greater than levels in the placebo group (52 plus minus 4 and 19 plus minus 3 mg dl(minus sign1), respectively). The data demonstrate that oral prednisone is effective in reducing proteinuria and improving the biological markers of disease activity (i.e., ESR, CRP, C3, C4, and anti-dsDNA) following short-term treatment of active lupus nephritis. Additionally, the more rapid change in anti-dsDNA levels and superior normalization of complement levels in the treatment group, although not statistically significant, are consistent with a biologic effect of misoprostol on lymphocyte function and the production of a pathogenic autoantibodies. PMID- 11854811 TI - A Potential Role for PGE and IL-14 (HMW-BCGF) in B-Cell Hyperactivity of Patients with Systemic Lupus Erythematosus. AB - Systemic lupus erythematosus (SLE) is a disease characterized by the prolonged production of high-affinity autoantibodies resulting in direct and immune complex mediated tissue damage. Because autoantibody responses occur over several years, memory B cells are likely to be involved. Interleukin-14 (IL-14) is a cytokine implicated in the generation and maintenance of normal memory B cells. Many of the actions of IL-14, including inhibition of antibody synthesis and upregulation of IL-14 receptors (IL-14R), are dependent on the formation of prostaglandin E (PGE) and subsequently cAMP. We observed that IL-14 induces phospholipase A(2) (PLA(2))-dependent release of arachidonic acid from phosphatidylcholine and phosphatidylinositol. Production of PGE is blocked by the PLA(2) inhibitor bromophenacyl bromide. Exogenous PGE (misoprostol) induces similar inhibition of antibody synthesis and increases in IL-14R as IL-14. Lymphocytes from patients with inactive SLE were noted to spontaneously produce PGE. Lymphocytes from normal donors produced PGE only after Sac-activation and IL-14 stimulation. Peripheral B and T lymphocytes from SLE patients, but not normal donors, spontaneously produced IL-14. Increased numbers of peripheral B lymphocytes from patients with inactive SLE expressed IL-14R, when compared to normal donors. Thus, increased production of IL-14 and PGE in SLE may result in expansion of a memory B-cell population capable of long-term autoantibody production. Further study will be necessary to confirm these preliminary findings and to examine in greater depth the regulation of PGE and IL-14 in SLE patients and normal donors. PMID- 11854812 TI - The Prostaglandin E(1) (PGE(1)) Analog Misoprostol Ameliorates Autoimmune Disease and Depletes T Lymphocytes in MRL-lpr/lpr Mice. AB - MRL-lpr/lpr mice spontaneously develop an autoimmune disease similar to human systemic lupus erythematosus (SLE). Disease manifestations include anti-DNA autoantibody production, arthritis, vasculitis, and an immune-complex glomerulonephritis. The development of autoimmune disease is associated with massive, generalized lymphadenopathy caused by accumulation of an abnormal T-cell population in peripheral lymphoid organs. In MRL-lpr/lpr mice, treatment with E series prostaglandins ameliorates renal disease and reduces peripheral lymphadenopathy. Little is known about mechanisms of action of E-series prostaglandins in murine lupus or the effects of these agents on other clinically important manifestations of SLE, such as arthritis and vasculitis. To further investigate the effects of an E-series prostaglandin in murine SLE, we administered the prostaglandin, E(1) (PGE(1)) analog misoprostol (1 mg kg(minus sign1) day(minus sign1)) to MRL-lpr/lpr mice for 8 weeks by twice daily subcutaneous injection. At 20 weeks of age, treatment with misoprostol reduced the severity of renal disease and arthritis but did not affect the extent or severity of vasculitis. The beneficial effects of misoprostol on arthritis and renal disease were associated with a significant decrease in splenic and lymph node weight in mice given the PGE(1) analog. This decrease in lymphoproliferation resulted primarily from a generalized reduction in the number of T cells in peripheral lymphoid organs. Thus, T-cell depletion was associated with beneficial effects on arthritis and nephritis in MRL-lpr/lpr mice, supporting a role for T lymphocytes in these disease processes. The ability of E-series prostaglandins to favorably modify autoimmune disease in this murine model suggests that misoprostol may be a useful adjunct to current therapies for the treatment of patients with SLE. PMID- 11854813 TI - Once-Daily Bedtime Dose of Roxatidine and Ranitidine in Acute Duodenal Ulcer: A Combined Assessment of Acid Inhibitory Activity and Healing Rate. AB - This study was carried out in order to compare the antisecretory effect of a single bedtime dose of roxatidine 150 mg and ranitidine 300 mg and to assess the relationship between the degree and the duration of acid suppression and the healing rates obtained in duodenal ulcer patients treated with the above regimens. Sixty-three patients with endoscopically proven ulcer underwent 24-h gastric pH-metry on day 0, day 1, and day 28 of treatment with both roxatidine and ranitidine. Ulcer healing was checked endoscopically after 4 weeks of therapy. RESULTS: Eight patients did not complete the study, leaving 55 patients eligible for final analysis, 28 in the roxatidine group and 27 in the ranitidine group. Duodenal ulcers were healed in 24--28 (85%) patients of the former and in 22--27 (81%) patients of the latter group (p minus sign NS). Gastric pH was significantly higher (p < 0.001) than basal values on days 1 and 28 with both H2 antagonists. The 24-h pH levels did not differ between day 1 and day 28 with both roxatidine and ranitidine. There was also do difference between the two active treatments. The pattern of gastric acidity significantly differed (p < 0.01) between responder (n = 46) and nonresponder (n = 9) patients to both H2-blockers, and this difference was mainly sustained by nocturnal pH. CONCLUSIONS: A bedtime close of roxatidine 150 mg and ranitidine 300 mg was able to heal more than 80% of duodenal ulcers within 4 weeks of treatment. The lack of tolerance to H2 blockers in duodenal ulcer patients contributes to this good result. The antisecretory effect of H2-antagonists is reduced in nonresponder patients with respect to responder patients and this is mainly due to an impaired control of nocturnal acidity. PMID- 11854814 TI - Pharmacokinetics of Ethanol in Arterial and Venous Blood and in End-Expired Breath during Vasoconstriction and Vasodilation. AB - We studied the influence of vasoconstriction and vasodilation on the concentrations and the pharmacokinetics of ethanol in arterial blood (a), venous blood (v), and end-expired breath (b). Ethanol 0.4 g kg(minus sign1) was given by intravenous infusion over 30 min to 12 healthy male volunteers, once in the morning and again in the afternoon the same day. During one infusion, the hand used to obtain venous blood was either cooled in water to 13--15 degreeC (N = 6 subjects) or warmed in a heating box at 60 degreeC (N = 6 subjects). Our results show that a--v differences and the pharmacokinetics of ethanol are sensitive to the constriction or dilation of blood vessels caused by local cooling or heating. Cooling of the hand doubled the a--v and b--v differences during infusion of ethanol, whereas warming reduced these differences by 10--80%. The a--b difference was positive during infusion but always became negative within 5 min after the infusion ended. However, cooling maintained the increased a--v difference for another 45 min after the infusion. Fitting of the data into a pharmacokinetic model consisting of a first-order distribution function and a zero-order (saturated Michaelis--Menten) elimination function showed that cooling altered the pharmacokinetic parameters describing the elimination of ethanol, whereas warming altered the parameters describing the distribution phase. PMID- 11854815 TI - Treatment of Obese Patients with Dexfenfluramine: A Multicenter, Placebo Controlled Study. AB - This randomized, placebo-controlled, double-blind study was designed to evaluate the safety and efficacy of dexfenfluramine (Dfen). Dfen 15 mg BID, and placebo were administered for 12 weeks to 337 moderately obese patients on calorically restricted diets. Patients were monitored for an additional 4 weeks. Efficacy was evaluated in 321 patients who were an average of 52% in excess of ideal body weight. Dfen-treated patients lost significantly more weight than did those treated with placebo (p less-than-or-equal 0.001). Small nonsignificant fluctuations in body weight were observed during the 4-week posttreatment period in both groups. The most common drug-related side effects were diarrhea, asthenia, dry mouth, and thirst (p less-than-or-equal 0.05 compared with placebo). Dexfenfluramine may become a valuable addition to weight loss and weight management programs. PMID- 11854816 TI - Phenobarbital in Sera of Epileptic Mothers and Their Infants. AB - The phenobarbital (PB) transition from epiletpic mothers to their breast milks and offspring in cases of PB monopharmacy with other antiepileptic drugs was investigated in 26 epileptic mothers and 24 offspring who were taking the breast milk from epileptic mothers. The mother's serum PB concentration in monopharmacy was almost the same in various stages (stage I, with 5 days after the delivery; stage II, 6--10 days after; stage III, 1--2 mothers after; stage IV, 3--5 months after). However, the PB concentrations in polypharmacy were markedly higher than those in monopharmacy. Concerning the PB concentration in breast milk, a significant increase was found in polypharmacy in comparison with monopharmacy only in stage I. On the other hand, the offspring's PB concentration in polypharmacy was markedly higher than that in monopharmacy, particularly in stage I. In another stages, PB concentration in monopharmacy gradually decreased according to the increase of stages. These results indicate that pharmacokinetics of antiepileptic drugs in the perinatal may be considerably different in PB monopharmacy and polypharmacy. PMID- 11854817 TI - The Therapeutic Implications of Left Ventricular Hypertrophy in the Hypertensive Patient. AB - Hypertension can be considered a syndrome broader than the condition of an increased blood pressure per se. Epidemiologic studies have established that high blood pressure is associated with an increased risk of cardiovascular events. Clinical trials of antihypertensive therapy have failed to show a consistent reduction in cardiovascular endpoints. The incidence of coronary disease has been reduced minimally, suggesting that factors beyond just measuring mm Hg in the hypertensive may be important in the genesis of atherosclerotic disease in hypertensive patients. Patients with hypertension appear to have an exaggerated vulnerability to the consequences of lipid abnormalities. In addition, hypertension is associated with insulin resistance and altered glucose tolerance. The increased plasma concentrations of insulin may produce proliferative effects on vascular smooth muscle and connective tissue, and these changes may adversely affect vascular integrity, leading to hypertrophy and facilitation of the arthersclerosis process. The left ventricle also appears to be involved in hypertension and this involvement may be independent of blood pressure. An increase in the muscle mass of the left ventricle as well as changes in its diastolic-filling characteristics occur early on with hypertension and may contribute to an adverse cardiovascular outcome. The arterial circulation is also involved. Alterations in structure or function of the vascular tree are reflected in diminished arterial compliance. These changes can be demonstrated prior to the appearance of clinical manifest hypertension. Based on these observations, treatment designed to reduce cardiovascular risk in hypertensive patients from cardiovascular events must not be based on blood pressure reduction alone but must take into account all the components of the hypertension syndrome. PMID- 11854818 TI - Synovial Tissue, Synovial Fluid, and Plasma Distribution of Nonsteroidal Anti inflammatory Drugs: Clinical Implications. AB - In the treatment of joint inflammation, the target tissue of nonsteroidal anti inflammatory (NSAID) drug action is the synovial membrane. Pharmacokinetic behavior of NSAIDs affects their concentration in tissue compartments. Ion trapping of NSAIDs (weak acids) in the more alkaline intracellular environment promotes cellular uptake in inflamed joints. Available data measuring synovial tissue concentrations of three NSAIDs demonstrates higher levels of oxaprozin in comparison to piroxicam and naproxen. PMID- 11854819 TI - The Investigators' Meeting: Obtaining Maximum Results. AB - This survey was undertaken to ascertain what clinical investigators liked and disliked about the meetings that precede the initiation of clinical trials and to determine if there are any differences in opinion between those investigators engaged in private practice and those affiliated with academic institutions. A 16 item multiple choice survey was mailed to 565 clinical investigators. Sixty-three percent of those polled returned the survey. A majority of respondents (67%) believes that attendance at investigators' meetings is beneficial to themselves and to their coordinators. Seventy percent prefer to attend a multicenter meeting in lieu of having an on-site initiation. Private practitioners prefer weekend meetings (59%) held at resorts (54%) where as academicians favored meetings to be held mid-week (42%) and were less unified as to the type of facility. The last choice for a meeting's location was the Northeast. More investigators prefer to make their own travel arrangement than to use a central travel planner. Both groups of investigators believe that the most beneficial parts of the meeting for themselves and for their coordinators are a formal presentation of the protocol and a discussion/answer session about the study. There was generalized agreement that the study's coordinator is the principal beneficiary of a discussion about the central laboratory. It is less clear who actually benefits from discussions of the drug brochure, FDA guidelines, case report form, etc. PMID- 11854820 TI - Smoking. PMID- 11854821 TI - The Antihypertensive Properties of the Angiotensin-Converting Enzyme Inhibitor Moexipril Given Alone or in Combination with a Low Dose of a Diuretic. AB - The antihypertensive characteristics of the angiotensin-converting enzyme inhibitor moexipril were evaluated in 413 patients with baseline setting diastolic blood pressures between 95 and 114 mm Hg. The study was double blind, with patients randomized to placebo or to differing doses of moexipril alone or in combination with a low dose of hydrochlorothiazide. Compared with placebo, moexipril 3.75 mg daily was not different, but single daily doses of 7.5, 15, and 30 mg were significantly more effective (as measured at trough, approximately 24 h after dosing) in decreasing the diastolic blood pressuring during an 8-week treatment period. The dose-response relationship indicated that no additional blood-pressure-lowering effect occurred above 15 mg daily. Hydrochlorothiazide 12.5 mg was not significantly more effective than placebo, but the combinations of the diuretic with moexipril doses of 3.75, 7.5, and 15 mg all produced significant antihypertensive actions. Interestingly, the 3.75-mg moexipril- hydrochlorothiazide combination was equally as efficacious as the higher doses. The combinations were all more effective than their respective moexipril and hydrochlorothiazide monotherapies. There were no meaningful laboratory changes except for decreased potassium concentrations in the patients on diuretic alone; this effect was attenuated in the low-dose moexipril combination. Only 14 of the 413 patients who entered the double-blind study period (3%) discontinued treatment because of adverse experiences. Thus, moexipril is a well-tolerated drug that has clear antihypertensive efficacy as a single agent in once-daily doses of 7.5--30 mg. When combined with hydrochlorothiazide 12.5 mg, it is effective in daily doses as low as 3.75 mg. PMID- 11854822 TI - The Effects of Age on the Pharmacokinetics and Pharmacodynamics of Cardiovascular Drugs: Application of Concentration-Effect Modeling. 1. Tolmesoxide. AB - The effects of age on the disposition and hemodynamic responses to the direct acting vasodilator tolmesoxide were examined in an integrated manner using a combined pharmacokinetic--pharmacodynamic (PKPD) modeling approach. Thirty subjects were studied to represent, as far as was possible, an age continuum. Single doses of tolmesoxide (100 mg) and placebo were administered intravenously and blood was withdrawn and heart rate and blood pressure were measured at frequent intervals up to 10 h post dose. No significant effects of age on the disposition of tolmesoxide and its sulfone metabolite were apparent. Tolmesoxide when compared with placebo produced significant changes in blood pressure and heart rate but PKPD modeling failed to demonstrate any significant effects of age on the blood pressure response to tolmesoxide. PMID- 11854823 TI - The Effects of Age on the Pharmacokinetics and Pharmacodynamics of Cardiovascular Drugs: Application of Concentration-Effect Modeling. 2. Acebutolol. AB - The effects of age on the disposition and hemodynamic responses to the selective beta(1) adrenoceptor antagonist were examined in an integrated manner using a combined pharmacokinetic--pharmacodynamic (PKPD) modeling approach. Thirty subjects were studied to represent, as far as was possible an age continuum. Single doses of acebutolol (20 mg) and placebo were administered intravenously and blood was withdrawn and heart rate and blood pressure were measured at frequent intervals up to 10 h post dose. No significant effects of age on the disposition of acebutolol and its major acetylated metabolite were apparent. Acebutolol when compared with placebo produced significant changes in blood pressure and heart rate and application of PKPD modeling demonstrated a significant negative correlation between blood pressure responsiveness to acebutolol and age. PMID- 11854824 TI - Effects of Age on the Pharmacokinetics and Pharmacodynamics of Cardiovascular Drugs: Application of Concentration-Effect Modeling. 3. Trimazosin. AB - The effects of age on the disposition and hemodynamic responses to the selective post-synaptic alpha(1) adrenoceptor antagonist trimazosin were examined in an integrated manner using a combined pharmacokinetic--pharmacodynamic (PKPD) modeling approach. Thirty subjects were studied to represent, as far as was possible an age continuum. Single doses of trimazosin (100 mg) and placebo were administered intravenously and blood was withdrawn and heart rate and blood pressure were measured at frequent intervals up to 10 h post dose. Based on regression analysis there was a statistically significant decline in the clearance of trimazosin with increasing age. In addition, based on the ratio of the AUC values for the major metabolite and parent drug, there was evidence of a decline in the relative clearance of matabolite with increasing age. Trimazosin when compared with placebo produced significant changes in blood pressure and heart rate that were statistically greater in elderly subjects. PKPD modeling revealed that both trimazosin and its metabolite 1-hydroxy trimazosin contributed significantly to the hemodynamic profile of the drug but the blood pressure responsiveness to both parent drug and metabolite were unaffected by age. Thus the greater response in the elderly subjects could be attributed to the decline in drug clearance with age. PMID- 11854825 TI - Effects of Aluminum/Magnesium Hydroxide and Calcium Carbonate on Esophageal and Gastric pH in Subjects with Heartburn. AB - This single-blind crossover trial compared the effects of single oral doses of two antacids on esophageal and gastric pH in subjects with heartburn. Gastric and esophageal pH were assessed in 83 subjects from 1 h before to 4 h after a refluxogenic meal. Subjects received two chewable tablets of a high-potency aluminum/magnesium hydroxide [Al(OH)3/Mg(OH)2] formulation (Mylanta Double Strength(TM)) or a calcium carbonate [CaCO3] formulation (Tums E-X(TM)), or placebo 1 h after the meal. Both antacid formulations significantly increased esophageal pH, as compared with placebo. Onset of action was faster with the Al(OH)3/Mg(OH)2 formulation than with the CaCO3 in 41 subjects, slower in 13 subjects, and identical in 29 subjects. Area under the esophageal pH--time curves after dosing were significantly greater for Al(OH)3/Mg(OH)2 than for CaCO3 (p < 0.05) and significantly greater for CaCO3 than for placebo (p < 0.05). The duration of Al(OH)3/Mg(OH)2 action in the esophagus was 82 min and 60 min for CaCO3 (p < 0.05). In the stomach, only Al(OH)3/Mg(OH)2 increased gastric pH compared with placebo. After ingestion of calcium carbonate, gastric pH usually remained at or below placebo values, a finding consistent with a calcium carbonate-induced "acid rebound." The duration of Al(OH)3/Mg(OH)2 action in the stomach was 26 min. These findings demonstrate the efficacy and relative superiority of this particular aluminum/magnesium hydroxide formulation compared with the calcium carbonate preparation at increasing esophageal and gastric pH. However, the magnitude and duration of action of both antacids on esophageal pH, in contrast to minimal effects on gastric pH, suggest strongly that the lower esophagus is the primary site of antacid activity in relief of heartburn. PMID- 11854826 TI - Effect of Gender and Menopausal Status on the Pharmacokinetics of Tirilazad Mesylate in Healthy Subjects. AB - The pharmacokinetics of tirilazad were assessed in men ages 40--60 years, women <40 years of age, premenopausal women ages 40--60, and postmenopausal ages 40- 60. Eight subjects in each group received single 3.0 mg kg(minus sign1) intravenous infusions of tirilazad mesylate over 10 min. Plasma concentrations of tirilazad and U-89678, an active metabolite, were measured by high-performance liquid chromatography. Tirilazad administration was well tolerated in all groups. Mean tirilazad clearance was 59.6% higher in young women compared to the middle aged men (35.6 plus minus 8.04 L h(minus sign1) vs. 22.3 plus minus 8.40 L h(minus sign1)). Mean tirilazad clearance in middle-aged women was 30.7% higher than in middle-aged men. Mean clearance in postmenopausal women (26.1 plus minus 4.21 L h(minus sign1)) was not significantly different than that in middle-aged men, but clearance corrected for body weight was significantly different between the men and postmenopausal women. Clearance in premenopausal middle-aged women (32.2 plus minus 7.60 L h(minus sign1)) was not significantly different from that in young women and was 44% greater than that in middle-aged men. Mean AUC(0minus signinfty infinity) and C(max) values for U-89678 were significantly higher in men than in all of the female groups. Among the women, values for U-89678 AUC(0minus signinfty infinity) were lowest in young women (467 plus minus 345 ng h ml(minus sign1), 8.8% of male value) and highest in postmenopausal women (1565 plus minus 1382 ng h ml(minus sign1), 29.4% of male value). The absolute values for U-89678 AUC(0minus signinfty infinity) must be interpreted with caution, as limited assay sensitivity and low plasma concentrations in the latter portion of the concentration-time profile in women precluded accurate determination of the terminal half-life and AUC(0minus signinfty infinity). Regardless, these results show that women, particularly premenopausal women, have lower concentrations of U 89678, an active metabolite of tirilazad, than are achieved in men. The gender differences in tirilazad and U-89678 pharmacokinetics are of sufficient magnitude that they may impact the clinical response of male and female patients to tirilazad treatment. PMID- 11854827 TI - The Effects of Lansoprazole on the Disposition of Antipyrine and Indocyanine Green in Normal Human Subjects. AB - This study was designed to evaluate the potential effects of acute and chronic daily oral doses of lansoprazole (60 mg) on the disposition of antipyrine, an almost completely metabolized low hepatic extraction compound, and indocyanine green, a hepatically secreted compound with high extraction ratio. The study utilized a randomized, placebo-controlled, double-blind, two-period crossover design. Sixteen of 18 subjects completed all phases of the study. Both antipyrine (10 mg kg(minus sign1)) and indocyanine green (0.5 mg kg(minus sign1)) were administered as single intravenous bolus doses on Days 1 and 7 of lansoprazole or placebo dosing. Acute exposure to lansoprazole had no statistically significant effects on the plasma pharmacokinetics of indocyanine green or antipyrine. After the seventh dose, there was a small but statistically significant reductions in indocyanine green total body clearance (CL), and elimination rate constant of 10.6% and 8%, respectively. Additionally, a small statistically significant reduction (8.6%) in antipyrine volume of distribution was detected. No other plasma antipyrine pharmacokinetic parameters were changed with concomitant lansoprazole administration. About a 12% increase in the recovery of one of the major antipyrine urine metabolites (NORA) was detected. Overall, this study demonstrates little or no effect of lansoprazole on the pharmacokinetics of antipyrine and indocyanine green. PMID- 11854829 TI - Clinical Therapeutic Conference: Sudden Death in a Young Athlete. PMID- 11854828 TI - An Introduction to Low-Molecular-Weight Heparins and Their Use in the Treatment of Deep Venous Thrombosis. AB - Low-molecular-weight heparins (LMWHs) are fractions of standard heparin (SH) with an average molecular weight of 4000--5000 daltons. They have the ability to inhibit the activity of factor Xa in the coagulation cascade but are unable to significantly change the activity of thrombin (factor IIa). Therefore, LMWHs have antithrombotic properties, but they do not prolong the activated partial thromboplastin time (APTT). In contrast to SH, the pharmacokinetic profiles of LMWHs are different. LMWHs exhibit minimal binding with plasma proteins, endothelial cells and platelet factor IV, a longer half-life, and linear elimination. Due to these properties, the clinical response is more predictable than that seen with SH. In addition, monitoring of clotting factors and subsequent dosage adjustment are not necessary to ensure efficacy when using a LMWH. These agents may have a decreased risk of thrombocytopenia and bleeding. LMWHs have been used in the initial treatment of deep venous thrombosis (DVT) and have been found to be safe and efficacious. When compared to SH, the LMWHs studied have been more effective in improving venographic scores and short-term treatment, but further evaluation is needed to determine if a benefit exists in preventing recurrent DVT. PMID- 11854831 TI - Introduction: Misoprostol and Eicosanoid Symposium. PMID- 11854830 TI - Misoprostol and Eicosanoids: A New and Exciting Area of Therapeutics. PMID- 11854832 TI - Overview: Misoprostol and Eicosanoids. PMID- 11854833 TI - Effect of Misoprostol on Acute and Chronic Inflammation. AB - Prostaglandins and their fatty acid precursors are important to regulation of cell function, and immune and inflammatory responses. Prostaglandin E compounds in particular have been shown to reduce inflammation and tissue injury. We examined the ability of misoprostol, the orally active analog of Prostaglandin E1, to influence inflammation in two animal models. In the subcutaneous air pouch model, acute inflammation was induced by tumor necrosis factor-alpha (TNF-alpha) and interleukin-1beta (IL-1beta). Misoprostol reduced pouch leukocyte counts in a dose dependent manner (33--49% v control), but did not alter fluid accumulation. The non-steroidal antiinflammatory agent, diclofenac, also reduced leukocyte counts in a dose dependent manner (28--66%) without affecting pouch fluid volume. Low dose misoprostol and low dose diclofenac used together reduced leukocyte counts by 39%, suggesting an advantageous snyergy in suppression of acute inflammation. Collagen induced arthritis (CIA) is a model of chronic inflammation. Misoprostol had no effect on the severity or course of CIA. Diclofenac reduced significantly all indices of inflammation tested, including joint swelling, number of affected joints and ability to walk. Misoprostol interfered with the antiinflammatory effect of diclofenac when the two compounds were administered together in the CIA model. These studies suggest that Misoprostol suppresses neutrophil mediated acute inflammation. PMID- 11854834 TI - The Role of Cyclooxygenase-2 in Inflammation. AB - Prostaglandins (PGs) can be synthetized via two isoforms of cyclooxygenase (COX). COX-1 is constitutively expressed in normal tissues, and its activity represent the normal physiological output of PGs. In inflammatory states, the newly discovered COX-2 is rapidly induced, and its activity accounts for the large amounts of PGs seen in inflammation. The commercially available nonsteroidal anti inflammatory drugs (NSAIDs) are nonselective inhibitors of both COX isoforms; therefore, they provide anti-inflammatory activity as well as side effects associated with COX-1 inhibition. Selective inhibition of COX-2 expression explains at least in part the potent anti-inflammatory activity of steroids. Anti inflammatory activity of newly developed COX-2 inhibitors, such as NS-398 or SC 58125, suggest a new approach of inflammatory diseases with more efficacious NSAIDs essentially devoid of side effects such as stomach ulcers. PMID- 11854835 TI - Preliminary X-Ray Investigations Into NSAID-Binding to Cyclooxygenase-1. AB - The prostaglandin H synthase structure reveals a fold similar to those of other heme peroxidases, but within this fold a second active site has evolved which catalyzes the cyclooxygenase reaction. The protein has also acquired two additional domains: a membrane-binding motif that mediates the protein's monotopic attachment to the membrane, and an epidermal growth factor-like module which is found just before the membrane-binding motif and is located in the dinner interface. PMID- 11854836 TI - Nitric Oxide Activates the Cyclooxygenase Pathway in Inflammation. PMID- 11854837 TI - Tyrosine Phosphorylation: Biochemical Events Involved in Cyclooxygenase II Activation by IL-1beta AB - Interleukin-1 induced the expression of cyclooxygenase II in renal mesangial cells. This effect was similar to that seen by phorbol myristate accetate which also induced the message for COX II. Cycloheximide superinduced the message for COX II in the presence of IL-1beta. While the message for COX II when stimulated by PMA was completely inhibited by dexamethasone with concentrations of dexamethasone which inhibited PGE(2) formation, dexamethasone only partially inhibited the message for COX II by Northern analysis. Genistein, an inhibitor of tyrosine phosphorylation completely inhibited the ability of IL-1beta to induce the COX II message. Immunocytochemical studies confirmed the ability of IL-1beta to selectively induce the COX II protein without any effect on COX I protein. These studies confirm the dependency of tyrosine phosphorylation in the IL-1beta induced expression of COX II message and protein. PMID- 11854838 TI - Misoprostol Modulation of Dendritic Cell Function in Immune Responses. AB - Dendritic cells (DC) from blood and other tissues are potent accessory cells for primary immune responses. Because prostaglandins from monocytes and macrophages can suppress DC and T-cell function, we sought to investigate the binding properties of misoprostol (MPL), a prostaglandin E(1) analog, and its regulation of DC function. Results of mouse and human experiments have suggested 1) that MPL could significantly inhibit DC-induced T-cell proliferation in oxidative mitogenesis and allogeneic mixed leukocyte reactions by decreasing interleukin-2 production in DC-T cell cocultures, and 2) that MPL could bind to human peripheral blood mononuclear cells via specific high-affinity MPL binding sites as well as through nonspecific MPL uptake. Taken together, these data suggest that MPL can bind high-affinity and/or nonspecific cell surface receptors and subsequently regulate T-cell growth and cytokine production such as that induced by DC and associated with primary immune responses. PMID- 11854839 TI - The Effect of Misoprostol on Arachidonic Acid Mobilization, Prostaglandin E(2), Production, and IL-1beta Signaling. AB - The effect of misoprostol on interleukin-1beta (IL-1beta)-mediated phospholipid metabolism, arachidonic acid release, and prostaglandin E(2) (PGE(2)) production was examined using normal skin fibroblasts and synovial fibroblasts from patients with osteoarthritis. We found that both IL-1beta and misoprostol induced arachidonic acid release, suggesting enhanced phospholipase A(2) activation. Both Il-1beta and IL-1beta/misoprostol, but not misoprostol alone, induced a significant increase in PGE(2) levels compared with controls. Even though PGE(2) production was not significantly increased by misoprostol alone, misoprostol synergistically enhanced IL-1beta-mediated cyclooxygenase activity (sixfold to eightfold) and PGE(2) synthesis in normal fibroblasts but not in OA synovial fibroblasts. Additionally, misoprostol dramatically affected the arachidonate labeling of triglyceride and cholesterol ester pools; the significance of this is as yet unclear. Together, the results suggest that misoprostol may upregulate the conversion of arachidonic acid to PGE(2) by enhancing the IL-1beta-induced activation/synthesis of cyclooxygenase. PMID- 11854840 TI - Differential Effect of PGE(2) on Adult and Neonatal T-Cell Dependent Immunoglobulin Secretion. AB - Prostaglandin E(2) (PGE(2)) and other cAMP elevating agents inhibited neonatal but not adult T cell dependent Ig production. PGE(2) did not inhibit IL-2 dependent anti-CD3 stimulated neonatal T cell proliferation or T cell-dependent neonatal B cell proliferation. The results could not be explained by differences in the effect of PGE(2) on cAMP production by neonatal and adult B and T cells or by a unique sensitivity of CD5+ B cells to inhibition by PGE(2). The enhanced sensitivity of neonatal T cell dependent Ig production to inhibition by PGE(2) may play a role in the increased susceptibility of neonatal B and T cells to tolerance induction. PMID- 11854841 TI - Regulation of Macrophage-Independent T-Cell Activation by Prostaglandins and Nonsteroidal Anti-Inflammatory Drugs. AB - We examined the hypothesis that both misoprostol (miso), a prostaglandin E(1) (PGE(1)) analog, and nonsteroidal anti-inflammatory drugs (NSAIDs) have significant and largely inhibitory effects on T-cell activation and, consequently, influence T-cell function. Studies were done using a macrophage independent system for T-cell activation by mitogenic combinations of immobilized anti-T-cell monoclonal antibodies (mabs), including combinations of anti-CD3 with anti-CD4 or anti-CD6. The results indicate that misoprostol, like other prostaglandins, can inhibit T-cell proliferation and the expression of mRNAs for cytokines critical in T-cell growth and immunoregulation. Somewhat higher concentrations of misoprostol are required than of PGE(1) to achieve comparable effects. However, even very low doses of misoprostol (0.01 &mgr;g ml(minus sign1)) may sometimes enhance T-cell activation. The effects on cytokine mRNA are not uniform; for a given dose and agent, one cytokine mRNA may be enhanced while another is unchanged or diminished. Whether a response is increased or decreased depends on several factors, including the dose of misoprostol or other PG tested, the strength of the response on which the agent is acting, and the level of preexisting activation or cytokine gene expression. Similar findings can be seen with treatment in vitro with NSAIDs, including sodium salicylate. When PGs (including misoprostol) and NSAIDs are combined, effects are often additive and sometimes appear synergistic, but paradoxical results can also occur, whereby each agent separately may be modestly inhibitory, but together they are stimulatory. The in vitro response of T cells to both PGs and NSAIDs (either separately or together) is, thus, highly complex. These data support the notion that these agents may have significant effects on the regulation of T-cell activation and function. PMID- 11854842 TI - Evaluation of Misoprostol and Analogs as Inhibitors of T-Cell Activation. AB - Prostaglandin E(2) (PGE(2)) is known to inhibit in vitro T-cell responses to mitogenic and antigenic stimuli. Interaction of PGE(2) with a G protein-coupled receptor activates adenylyl cyclase, leading to cAMP formation and inhibition of interleukin-2 (IL-2) production and T-cell proliferation. Despite these effects, the application of PGE(2) as an anti-inflammatory agent has been compromised by its unfavorable pharmacodynamic and side-effect profile. Because of the potential utility of synthetic analogs as prostaglandin-based therapeutics, we evaluated the effect of misoprostol and over 100 structural analogs on cAMP formation and T cell activation. Our results indicate that micromolar concentrations of misoprostol and particular analogs elicited a rapid and substantial rise in cAMP levels in human peripheral blood mononuclear cells. Analogs which increased cAMP also suppressed IL-2 production and T-cell growth in vitro, whereas those devoid of suppressive activity weakly induced nucleotide synthesis. Despite extensive chemical alteration of the prostanoid structure, no single analog was superior to misoprostol in inducing cAMP or modulating T-cell activity. Misoprostol and suppressive analogs were also evaluated in vivo in a murine model of antigen induced T-cell proliferation. Prostaglandins, administered at maximum tolerable doses, were ineffective in blocking a T-cell response to alloantigenic stimulation, whereas cyclosporine and prednisolone were potent inhibitors of this response. Overall, our results indicate that misoprostol and related analogs suppress T-cell activation in vitro but require concentrations 1000-fold greater than the low nanomolar plasma levels achieved with clinical doses of misoprostol. Whether misoprostol analogs of sufficient potency can be developed for pharmacologic attentuation of T-cell activation in vivo remains to be determined. PMID- 11854843 TI - Interleukin-6 Gene Expression by Prostaglandins and Cyclic AMP Mediated by Multiple Regulatory Elements. AB - We have evaluated the potential role of prostaglandins and their second-messenger Cyclic Adenosine Monophosphate (cAMP) in the activation of interleukin-6 (IL-6) promoter regulatory elements leading to IL-6 expression in monocytic cells. We demonstrate that prostaglandins of the E series and their second-messenger cAMP induce the IL-6 promoter in the murine monocytic cell line PU5-1.8. Stimulation with both cAMP and LPS results in a marked synergistic effect. We show that the endogenous IL-6 gene is induced by cAMP as well, even though to a lesser extent than by LPS, suggesting distinctive effects of cAMP and LPS on posttranscriptional events. Mutations eliminating potential transcription factor binding sites, including the multiple-response element (MRE), AP-1, NF-IL6, and NF-kappaB binding sites, significantly reduce, but do not completely abrogate, inducibility by cAMP or prostaglandin E1, whereas alterations of four additional putative regulatory elements have no effects. In contrast, LPS-induced promoter activity is almost completely abolished by mutations in the NF-kappaB binding site, suggesting that a single regulatory element is crucial for inducibility by LPS, whereas no individual element is absolutely essential for cAMP signaling. Induction of the AP-1, NF-IL-6, and NF-kappaB elements by cAMP is correlated with the appearance of inducible factors binding to these sites, whereas factors binding to the MRE are constitutively expressed. Our results suggest that cAMP and prostaglandins act through multiple, partially redundant, regulatory elements to induce IL-6 expression in monocytic cells. PMID- 11854844 TI - The Basic Amino Acid-Rich DNA-Binding Element of the NF-IL6 Transcription Factor Contains Two Functionally Distinct Subdomains. AB - Nuclear factor-interleukin-6 (NF-IL6), a human transcription factor of the CCAAT box/enhancer binding protein (C/EBP) family, is widely implicated as a "master regulator" of hepatic acute-phrase response and inflammatory cytokine gene expression. NF-IL6 contains, at its N-terminus, an alanine- and proline-rich transactivation domain, followed at the C-terminus by a basic domain-leucine zipper (bZIP) DNA-binding motif. Understanding how the NF-IL6 transcription factor interacts with DNA is fundamental to its role as a transactivator because sequence-specific DNA-binding is prerequisite to promoter activation. We review our findings on the identification of the minimal "core" DNA-binding domain and the discovery of a novel bZIP element that influences DNA-binding kinetics. Manipulation of this domain allows for design of molecules that can be used as competitive antagonists or targeted delivery of molecules to selected regions of the eukaryotic genome. PMID- 11854845 TI - Repression of Constitutive and Interferon-gamma-Induced Class II Major Histocompatibility Complex Genes by Cyclic AMP and Prostaglandins. PMID- 11854846 TI - Molecular Dissection of the Murine IL-1beta Promoter. AB - Interleukin-1 (IL-1) is involved in a broad range of biological activities that affect immunological, inflammatory, and nonimmunological responses. Although the role of the IL-1 proteins in normal physiological responses in vivo remains incompletely defined, there is substantial evidence that excessive production of IL-1 contributes to the pathogenesis of many illnesses with autoimmune or inflammatory components, including rheumatoid and osteoarthritis, type I diabetes mellitus and atherosclerosis. Despite numerous reports on IL-1 regulation, very little is known regarding the molecular details of IL-1 production, particularly at the transcription level. This review will focus on our studies of transcriptional regulation of the murine IL-1beta gene and, where appropriate, comparison to similar studies of the human IL-1beta gene. A basic understanding at this level should lead to effective pharmacological intervention and, ultimately, to control of inflammatory disease. PMID- 11854847 TI - Prostaglandin E Suppresses Hepatic Fibrosis: Section I. The In Vivo Approach; Section II. The In Vitro Approach. AB - Recent studies suggest that prostaglandin E may have the ability to suppress cytokine responsiveness. We examined the effects of prostaglandin E administration on several parameters of the acute and chronic hepatic injury induced by bile duct ligation. Enisoprost, a prostaglandin E(1) analog was found to suppress early hepatic and Ito cell type I collagen gene expression without diminishing the induction of the fibrogenic cytokine, transforming growth factor beta. Overall hepatic inflammation and cell proliferation were not altered, suggesting that prostaglandin E acts distal to the initial injurious event(s). During the later phases, drug administration reduced total collagen accumulation as well as type I collagen periductular infiltration associated with early nodule formation. Ito cell mitogenesis occurs during liver injury and fibrogenesis in vivo coincident with the de novo expression of Ito cell platelet-derived growth factor beta (PDGFbeta) receptor messenger RNA. PDGF-induced mitogenesis was studied in cultured rat hepatic Ito cells which resemble the myofibroblast associated with liver injury. Pretreatment with prostaglandin E markedly suppressed the PDGF response in a dose-dependent fashion. The PDGF-induced cascade was studied plus minus PGE to determine the level of regulation which induced the observed suppression. PGE caused no apparent diminution in the abundance of the surface PDGFbeta receptor nor its subsequent activation and tyrosine phosphorylation following PDGF stimulation. The cytoplasmic "secondary messengers" mitogen-activated protein kinase pp42--44 and raf kinase appeared to be comparably induced and therefore unaffected by PGE. Raf perinuclear translocation was also intact, and comparable degrees of nuclear egr, fos, and jun expression occurred. Because other studies have suggested that many of these features of the PDGF cascade may be causally and sequentially linked, the data collectively suggest that the dominant PGE mitogenic suppressive effect resides at a Raf-MAP parallel pathway or at a nuclear level distal to the induction of these early growth response genes. PMID- 11854848 TI - Stimulation of Lipogenesis by Interleukin-6 and Misoprostol-Free Acid in Isolated Rat Hepatocytes. AB - Recent work has established that Kupffer cell products, including prostaglandins, can act directly on hepatocytes to modify glucose and lipid metabolism. Additionally, prostaglandins can act on Kupffer cells to modify the expression of cytokines. Interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) stimulate hepatic lipogenesis following in vivo administration. To define the direct effects of these cytokines on lipogenesis in primary culture rat hepatocytes, hepatocytes were cultured in the presence of IL-6 or TNF-alpha for periods of 24--72 h. IL-6 caused an increase in hepatocyte lipogenic capacity (56% increase by 12.5 ng ml(minus sign1) IL-6 after 72 h of cytokine exposure). The increase in cellular lipogenic capacity was confirmed using (3)H2O as the radiotracer. TNF-alpha did not increase the rate of hepatocyte lipogenesis. Neither IL-6 nor TNF-alpha modified rates of lipogenesis upon acute exposure to the cytokine. Misoprostol-free acid (0.1 &mgr;M) acutely increased hepatocyte lipogenic rates by 14% in the presence of glucagon. These results demonstrate that IL-6 can act directly on hepatocytes to induce lipogenic capacity and that E series prostaglandins can antagonize the acute inhibition of lipogenesis by glucagon. Because IL-6 is produced by Kupffer cells, and its expression is modulated by prostaglandins, the Kupffer cell is a novel target for prostaglandin therapy. Administration of prostaglandins may provide a novel strategy for pharmacologic therapy of disorders of glucose or lipid metabolism. PMID- 11854849 TI - Eicosenoids Modify Experimental Allergic Encephalomyelitis. AB - Experimental allergic encephalomyelitis (EAE) is an autoimmune inflammatory disease of the brain and spinal cord. It is an animal model of postinfectious encephalomyelitis and multiple sclerosis (MS). In EAE and in MS, monocytes and Th1 lymphocytes penetrate the blood-brain barrier, and the ensuing inflammation causes demyelination and death of oligodendroglia. PGE is a product of blood Mo and of brain glial cells that affects immune regulation. PGE and other cAMP agonists inhibit monocyte function and secretion of cytokines by Th1 cells. However, they have minimal effects on some cytokines secreted by Th2 cells. We hypothesized that eicosenoids would inhibit central nervous system inflammation mediated by Th1 cells. We found that misoprostol, a long-acting PGE1 analog, inhibited clinical and histological signs of moderately severe EAE in Lewis rats. Indomethacin also suppressed EAE and enhanced the LAPGE effect. Both agents suppressed EAE when administered either from the time of immunization or from the onset of clinical disease. The combination of misoprostol and indomethacin inhibited delayed-type hypersensitivity reactions to MBP (a Th1 response). These agents also inhibited in vitro lymphocyte proliferation to mitogens and MBP. Leukotrienes (LKT) elevate intracellular cGMP and amplify immune responses, the opposite of cAMP agonists. We found that LKT synthesis inhibitors blocked EAE, presumably by lowering levels of cGMP in inflammatory cells. Reduction of LKT synthesis enhanced the effects of misoprostol plus indomethacin on EAE. PGE analogs, indomethacin, and inhibitors of LKT synthesis block autoimmune responses to brain antigens in vitro and in vivo. Modification of intracellular cAMP and cGMP levels with these agents may ameliorate inflammatory and autoimmune diseases. PMID- 11854850 TI - Effect of Prostaglandin E on the Humoral Response to a Specific Antigenic Stimulus in Human Lymphocytes. AB - Human lymphocytes from individuals immunized with tetanus toxoid >1 year previously that are cultured for 10 days with tetanus toxoid in vitro produce anti-tetanus toxoid antibody without a polyclonal antibody response. This specific anti-tetanus toxoid antibody response is augmented by the addition of prostaglandin E(2) to the cultures. Removal of CD8(+) cells prior to culture results in increased antibody production, but does not eliminate the augmentation by prostaglandin E(2). Thus, prostaglandin E(2) stimulates specific antibody production not mediated via inhibition of suppressor cell function. PMID- 11854851 TI - Misoprostol Stimulates cAMP Generation in Human Leukocytes: Synergy with Colchicine Suggests a New Potential for Established Drugs. AB - In isolated leukocytes, elevation of cAMP can inhibit various proinflammatory and immune functions. The prostaglandins E (PGEs) are known to stimulate leukocyte cAMP production, and for years they have been viewed as potential immunosuppressive and/or anti-inflammatory agents. However, their clinical use is severely limited by extreme metabolic instability and by poor oral absorption, which necessitates administration by infusion or injection. Misoprostol is a synthetic analog of PGE(1) that is relatively stable and orally absorbable. We examined the effects of misoprostol on cAMP production in leukocytes, in view of the possibility that it mimics PGE(1) and, thus, might represent a clinically useful immunosuppresive or anti-inflammatory drug. Our results indicate the following: (1) Misoprostol increases leukocyte cAMP production in a dose dependent manner (similar20 nM to >100 &mgr;M) and acts by stimulating adenylate cyclase. (2) Its potency and maximal effect are somewhat less than those of PGE(1) (3) cAMP generation in response to either misoprostol or PGE(1) is transient (in the presence of isobutylmethylxanthine to inhibit endogenous phosphodiesterases). (4) Misoprostol's stimulation of adenylate cyclase is synergistically increased by pretreatment of cells with colchicine, a microtubule disrupting agent that is currently used for prophylaxis and treatment of gout. (5) Colchicine acts by increasing the initial rate of cAMP production and not by prolonging the response to misoprostol. (6) A clinically relevant dose of colchicine (0.25 &mgr;M) is effective given sufficient pretreatment time. (7) Whereas a clinically relevant dose of misoprostol (3 nM) is ineffective alone, preexposure of cells to colchicine enables such a dose to stimulate cAMP generation significantly. The combination of misoprostol with colchicine might eventually prove useful in the therapy of immune or inflammatory disease. PMID- 11854852 TI - Three Rs potential in the development and quality control of pharmaceuticals. AB - The intention of a pharmaceutical company is to develop new, efficient products quick and with a minimum of costs. Compared to in vitro methods, animal experiments in general consume much more time and resources (costs as well as time to the market) than in vitro methods. Therefore, the use of whole animal models depends primarily on the judgement of their efficacy in the screening process, but the willingness to incorporate in vitro methods in general is high and is furthered by new developments such as high-throughput screening. Nevertheless, in vitro tests might be politically promoted by increasing their costs (quality controls, requested housing conditions) and duration (time to start of an experiment, sequential performance). Which models are favoured by industry to include them in a screening process: They have to be based on our most recent understanding of the respective disease, well characterised to allow interpretation of results and require only limited development time. All these aspects argue in favour of collaboration between industry and academia, where our understanding of pathophysiology is generated and mechanism based models are developed and characterised. However, technology transfer towards industry represents a bottle-neck for industrial use of these new in vitro models. New platforms to promote this transfer should be developed in order to bring together developer and user of novel in vitro systems and promote demonstration projects. Financing of such collaborations is not the key problem (the development of a single drug makes up to 500 million $) but the dilemma of publication of results: The development advantage compared to competitors depends on the exclusive use of novel models. The protection of intellectual property rights and the public interest in spreading alternatives to animal experiments must be balanced, e.g. by delayed but indispensable publication or advantages for companies employing alternatives in the regulatory approval process for a new drug. Quality control of therapeutic drugs (except hormones and blood products) represents a minor field of animal consumption with the exception of pyrogenicity testing. Despite considerable progress due to the introduction of the Limulus assay which represents the most successful in vitro alternative in use so far. However, some limitations of this in vitro test might be overcome in the near future by the currently validated human whole blood assay. During the last few years considerable progress has been made in the replacement (and deletion) of animal tests required for the potency and safety testing of hormones. This has been made possible by biotechnical production methods, by better-defined products, and because physico-chemical methods can be used for the potency testing of these products. In general, the better defined a drug is, the easier chemical, physical or in vitro techniques can be used for batch control. Control authorities should therefore urge the use of highly standardised components. PMID- 11854853 TI - Three Rs potential in the development and quality control of immunobiologicals. AB - Immunobiologicals (vaccines, immunoglobulins and -sera) are considered to be the most cost-effective tools in the prevention of infectious diseases. Their importance will further increase due to various eradication programmes of the WHO and EU and the emergence of new infectious diseases or the re-emergence of diseases as diphtheria and tuberculosis. The production and quality control of immunobiologicals are regulated by monographs and guidelines, which are issued by international or national Pharmacopoeias (e.g. Ph. Eur.), international organisations (e.g. WHO, O.I.E.) and international regulatory bodies (e.g. EMEA). Their purpose is to assure the quality of the product, i.e. its safety and potency. It is estimated that 10 millions of laboratory animals are world-wide used for the production and quality control of immunobiologicals, of which 80% are needed for the safety and potency testing of the finished product (batch control). In recent decades, the use of Three Rs principles has been recognised by the above mentioned organisations and various national competent authorities and been incorporated into general monographs and guidelines. Several tests with questionable relevance have been deleted from Ph. Eur. monographs (e.g. abnormal toxicity test, extraneous agents testing of viral vaccines for carnivores) or are now carried out during production. Reduction of the number of animals used could be achieved by introducing single-dilution tests. A large number of immunochemical tests have been developed, which could completely or partly replace the use of animals for potency testing, however, only a few have been validated so far (e.g. ToBI and ELISA for potency testing of human and veterinary tetanus vaccine; ELISA for potency testing of erysipelas vaccine). Regulatory acceptance of validated alternative methods is still a critical step. In particular, the period between successful validation and the implementation appears to be far too long. Reasons for this could be the slow process of multinational agreement to revise pharmacopoeial monographs and guidelines, and the time-consuming and expensive production of sufficient reference material (antigen, sera etc) for the new test systems. The shift in the quality control concept from reliance on final batch testing to the concept of consistency of production offers the opportunity to reduce the numbers of animals being used and promote the use of alternative methods. Emphasis is put on a combination of in vitro tests, which could make it possible to monitor batch-to-batch consistency. This new concept of quality control is already in place for the new well-defined vaccines. In most cases, non-animal methods are used for monitoring consistency at critical steps in the production and testing of a vaccine. Whether the concept of consistency of production could be also applied to the conventional, less defined products, should be investigated. Only little progress has been achieved with regard to international harmonisation. Most of the manufacturers produce for the world market, so harmonisation of the requirements or mutual recognition of tests would help to reduce the use of animals. There is agreement that for the time being animals will still be needed for the development of vaccines in order to gain best knowledge on the disease, the pathogen and the specific immune response, including: pathogenesis, identification of the protective antigens, the way the antigen is processed, the dynamics of the immune response, the induction of memory, and the selection of the best adjuvant. With regard to routine batch release of conventional products, a number of Three Rs approaches are already available and should further be developed and validated. Whereas routine batch release of new products should be based on in vitro methods already established during their development. PMID- 11854854 TI - [Vitamin and zinc supplementation are useful in age-related macular degeneration]. PMID- 11854855 TI - [Polymers in ocular drug formulations]. PMID- 11854856 TI - [ADAL-1(R): a new synthetic adhesive for use in strabismus surgery]. PMID- 11854857 TI - [Impression cytology contribution to differential diagnosis of Sjogren syndrome in the ophthalmological clinic]. AB - PURPOSE: First, to assess the use of impression cytology, clinical and laboratory tests in the daily clinic for the early diagnosis of Primary Sjogren Syndrome (SS1), and second, to establish morphological differences between SS1, keratoconjunctivitis sicca (KCS) and healthy controls using impression cytology. MATERIAL AND METHOD: The study comprised 35 patients suffering from primary Sjogren syndrome, same number of patients with keratoconjunctivitis sicca and normal healthy subjects. All groups underwent clinical tests (Schirmer's test, break-up time, and bengal rose staining), laboratory tests (tear protein pattern, tear osmolarity and serum immunoglobulin readings), and impression cytology on different areas of the conjunctiva and cornea. RESULTS: All patients presented one or more subjective symptoms of ocular irritation although seeming normal at naked eye. Two patients with Sjogren's syndrome showed total corneal keratinization. Clinical and laboratory tests did not show significant differences between Sjogren's syndrome patients group and keratoconjunctivitis group. Serum tests of antibodies and immunoglobulins did present diagnostic values. Impression cytology showed significant differences between Sjogren syndrome patients compared to the rest of groups with regard to epithelial and goblet cells. CONCLUSIONS: Impression cytology consists in a hystopathological test and it is considered the only ophtalmological test that offers a differential diagnosis in Sjogren's syndrome and should be included in a classification criteria. Inpression cytology presents significant differences between Sjogren syndrome and keratoconjunctivitis sicca. PMID- 11854859 TI - [Selective ophthalmic artery fibrinolysis in acute central retinal artery occlusion]. AB - PURPOSE: To evaluate efficacy and reliability of local intra-arterial fibrinolysis (LIF) in Retinal Central Artery Occlusion (RCAO). MATERIAL AND METHOD: We have studied 12 patients with RCAO. Seven patients (group I) were treated with conventional therapy and 5 patients (group II) underwent treatment with urokinase via transcutaneous femoral catheter into the ophthalmic artery. Thereafter patients received heparin sulfate for 3 days. RESULTS: Mean follow up was 14 months and mean time from onset of symptoms to the beginning of therapy was 11 hours. One patient in group I showed improvement of visual acuity (from counting fingers to 0.1). In all patients in group II there was angiographic evidence of improved perfusion of retinal arteries and 4 patients (80%) showed improvement in visual acuity. The best results were obtained when LIF was performed before 10 hours after onset of symptoms. No complications were observed after the fibrinolysis treatment. CONCLUSION: LIF in ophtalmic arteries for the treatment of RCAO reduces dosage of fibrinolytic agents and becomes a safe and useful treatment during the first hours post RCAO. PMID- 11854858 TI - [A study on developmental characteristics of optic nerve in a rat model of materno-foetal hypothyroidism]. AB - PURPOSE: It is well known that vertebrates Central Nervous System reacts to a wide variety of hormones, and growth factors. However the basic maturation mechanisms and remodelling processes remain in general unknown. Thyroid hormones tri-iodothyronine (T(3)) and thyroxine (T(4)) modulate this metabolism, playing a role in cell differentiation and proliferation and in gene expression during growth. The aim of the present study was to investigate whether or not low levels of thyroid hormone in blood, obtained with an experimental model of congenital neonatal hypothyroidism referred to herein, may induce changes in optic nerve development, mainly in processes of macroglial cell genesis and myelination. MATERIAL AND METHOD: We used an experimental model of materno-foetal hypothyroidism (MFHP) set up in a rat through chemical thyroidectomy. A solution of methyl-mercapto-imidazole and KClO(4) was administered with drinking water to pregnant rats and their offspring during gestation and lactancy. A group of rats was maintained in parallel as controls (C-G). Optic nerves were excised from both groups at key postnatal developmental stages (P) and these were prepared for light and transmission electron microscopy. RESULTS: Cross-sectional areas of optic nerves in the MFHP-G group appeared significantly smaller compared to C-G, this, during the perinatal phase as well as P25 (p<0.01). Macroglial cell nuclear cross-sectional areas showed increasing values in both groups. Data from MFHP-G, however, showed significantly lower than those of C-G (p<0.01). Between 4 to 6 days delayed myelination was at all times observed in the treated group. CONCLUSIONS: All results suggest that T(3) and T(4) regulate optic nerve development by stimulating glial cells function at cell nuclear level, probably through specific receptor binding sites, as suggested in earlier literature (Pinazo-Duran et al. Arch. Soc. Esp. Oftalmol., 1997). PMID- 11854860 TI - [Topographic classification of glaucomatous visual fields]. AB - PURPOSE: To propose a classification of glaucoma visual fields based on affected ganglion cells axons on their way through the optic nerve head. MATERIAL AND METHOD: 255 Octopus 1-2-3 visual fields from glaucoma patients and glaucoma suspected patients were analyzed. Determination coefficients (r(2)) between the 10 points closest to the blind spot and the 73 reminding ones were calculated by linear regression. RESULTS: Significant correlation was found (r(2)>0.33) with very distant points sensitivity related to the ones closer to the optic disc due to the fibers path. Seven areas could be defined by this method: three of them are not affected or latterly affected by the disease: S1 and I1 corresponding to the upper and lower papillo-macular bundle and T, located on the temporal aspect of the blind spot. Areas S2 and L2 correspond to the nasal and para-central upper and lower sensitivities. Areas S3 and I3 are in the border of upper and lower nasal fields where axons enter the optic disc at the apical portion. CONCLUSIONS: There is a relationship between the anatomical arrangement of ganglion cells axons on their way to the optic nerve and topography of glaucoma defects which may justify this proposed classification. PMID- 11854861 TI - [Functional improvement after vision rehabilitation in low monocular vision after myopic macular degeneration and retinal detachment]. AB - PURPOSE: We present a case of a 62 year-old woman, with a single eye functional vision (VA of 0.16) who improved her vision after following a vision rehabilitation program, which included optical and non-optical devices for daily performance. CONCLUSIONS: A correct optometric evaluation and a training program are key factors to improve quality of life in low vision patients, whose ocular pathology allows no other treatment. PMID- 11854862 TI - [Macular translocation in myopic patient wearing a phakic intraocular lens]. AB - PURPOSE/MATERIAL AND METHOD: To report a case of macular translocation surgery in a myopic patient with a phakic intraocular lens, which developed subfoveal choroidal neovascularization. RESULTS: A 45 year-old woman wearing a phakic intraocular lens to correct a high myopia presented a 0.3 visual acuity because of subfoveal choroidal neovascularization. Peripheral retina had been treated by argon laser for lattice degeneration. Limited macular translocation was performed. Chorioscleral infolding was created by a circular scleral buckle. Her visual acuity, six months after surgery became 0.7. CONCLUSION: This report shows the feasibility of the use of limited macular translocation for the management of high myopic eyes with phakic intraocular lenses, in patients who suffer from subfoveal choroidal neovascularization. PMID- 11854863 TI - [Adjustable sutures in blepharoptosis surgery]. AB - PURPOSE/MATERIAL AND METHOD: Current adjustable suture techniques are analysed and new variants are described. RESULTS/CONCLUSION: Adjustable sutures applied to tarsal fixation of the elevator muscle and shortening-enlargement of the frontal suspension strip are presented. In the both techniques, after three to four days, the adjustment and the final fixation are performed. PMID- 11854864 TI - [A case of primary toxoplasmosis in an immunocompetent patient]. AB - CASE REPORT: A case of an immunocompetent patient presenting primary systemic toxoplasma infection involving the eye (condition seen in less than 3% of primary infections). The patient showed reactivation of this primary focus two years later. DISCUSSION: Diagnosis of toxoplasm retinitis is based on a typical lesion consisting in an area of active retinitis adjacent to an inactive corioretinal scar. Differential diagnosis must consider other causes of retinal coroiditis in primary infection cases: sarcoidosis, tuberculosis, syphilis as well as viral and fungal infections. Ocular toxoplasmosis was confirmed by serum tests. PMID- 11854865 TI - [A. Mooren and the sympathetic vision conditions (II)]. PMID- 11854867 TI - A polymorphism in the beta1 adrenergic receptor is associated with resting heart rate. AB - Resting heart rate is significantly associated with cardiovascular morbidity and mortality. However, the extent to which resting heart rate is genetically determined is poorly understood, and no genes have been found that contribute to variation in resting heart rate. Because signaling through the beta1 adrenergic receptor is a key determinant of cardiac function, we tested whether polymorphisms in this receptor are associated with resting heart rate. A cohort of >1,000 individuals of Chinese and Japanese descent, from nuclear families, was genotyped for two polymorphisms, resulting in a serine/glycine substitution at amino acid 49 (Ser49Gly) and an arginine/glycine substitution at residue 389 (Arg389Gly), in the beta1 adrenergic receptor. For comparison, polymorphisms in the beta2 and beta3 adrenergic receptors were also evaluated. The Ser49Gly polymorphism was significantly associated (P=.0004) with resting heart rate, independent of other variables, such as body-mass index, age, sex, ethnicity, exercise, smoking, alcohol intake, hypertension status, and treatment with beta blockers. The data support an additive model in which individuals heterozygous for the Ser49Gly polymorphism had mean heart rates intermediate to those of either type of homozygote, with Ser homozygotes having the highest mean heart rate and with Gly homozygotes having the lowest. Neither the Arg389Gly polymorphism in the beta1 adrenergic receptor nor polymorphisms in the beta2 and beta3 adrenergic receptors were associated with resting heart rate. The heritability of heart rate was 39.7% +/- 7.1% (P<10-7). PMID- 11854868 TI - Homozygosity for multiple contiguous single-nucleotide polymorphisms as an indicator of large heterozygous deletions: identification of a novel heterozygous 8-kb intragenic deletion (IVS7-19 to IVS15-17) in a patient with glycogen storage disease type II. AB - Current methods for detection of mutations by polymerase chain reaction (PCR) and sequence analysis frequently are not able to detect heterozygous large deletions. We report the successful use of a novel approach to identify such deletions, based on detection of apparent homozygosity of contiguous single-nucleotide polymorphisms (SNPs). The sequence analysis of genomic DNA PCR products containing all coding exons and flanking introns identified only a single heterozygous mutation (IVS18+2t-->a) in a patient with classic infantile-onset autosomal recessive glycogen storage disease type II (GSDII). Apparent homozygosity for multiple contiguous SNPs detected by this sequencing suggested presence of a large deletion as the second mutation; primers flanking the region of homozygous SNPs permitted identification and characterization by PCR of a large genomic deletion (8.26 kb) extending from IVS7 to IVS15. The data clearly demonstrate the utility of SNPs as markers for large deletions in autosomal recessive diseases when only a single mutation is found, thus complementing currently standard DNA PCR sequence methods for identifying the molecular basis of disease. PMID- 11854869 TI - How much cardiac output is enough? PMID- 11854870 TI - Is clinical assessment of the circulation reliable in postoperative cardiac surgical patients? AB - OBJECTIVE: To study adult patients in the immediate postoperative period after cardiac surgery and determine whether cardiac output (CO) and systemic vascular resistance (SVR) are generally within normal limits and whether clinical assessment of CO provides an adequate approximation to guide treatment. DESIGN: Prospective study. SETTING: Overnight intensive recovery unit, St Thomas' Hospital, London. PARTICIPANTS: Fifty patients who had undergone cardiac surgery within the previous 5 hours and whose COs were not being measured. INTERVENTIONS: The physician (if present) or nurse managing the patient was asked to make a clinical estimate (as low, normal, or high) of CO and of SVR. CO was measured by lithium dilution. The clinically estimated values were compared with the measured values, after adjusting for patient size. MEASUREMENTS AND MAIN RESULTS: In 29 of 50 (58%) patients, cardiac index (CI) (20 of 50 [40%] patients) or systemic vascular resistance index (SVRI) (22 of 50 [44%] patients) was outside normal limits (CI, 2.7 (plus minus 30%) L/min/m(2); SVRI, 2,500 (plus minus 30%) dyne center dot sec center dot cm(minus sign 5)/m(2)). When CI was outside the normal range, the clinical estimate was usually wrong (13 of 20 [65%]); when SVRI was outside the normal range, the clinical estimate was also usually wrong (16 of 22 [73%]). CONCLUSIONS: In the early postoperative period after cardiac surgery, clinical assessment of the circulatory state is frequently misleading, and there should be a low threshold for measuring CO. PMID- 11854871 TI - Equivalence of bioimpedance and thermodilution in measuring cardiac index after cardiac surgery. AB - OBJECTIVE: To compare thoracic electrical bioimpedance (TEB) cardiography versus pulmonary artery thermodilution (TD) derived cardiac index in patients after cardiopulmonary bypass. DESIGN: Prospective, blinded electronic data collection. SETTING: Intensive care unit of a military hospital. PARTICIPANTS: Post cardiopulmonary bypass patients for primary comparison between technologies (n = 20) and patients for comparison of variability within each technology (n = 20). INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Cardiac index values by TEB or TD were collected simultaneously. Linear regression, Lin's concordance correlation coefficient, bias, and precision measures within the large data set group and within each patient over time were calculated. Linearity in regression and Lin's concordance correlation coefficient of 0.99 were shown. A bias of 0.07 L/min/m(2) and precision of 0.40 L/min/m(2) were within acceptable clinical limits, as were equivalence test results. CONCLUSIONS: TEB is equivalent to TD derived cardiac index in postoperative cardiac surgery patients. PMID- 11854872 TI - Epidural anesthesia and analgesia: effects on recovery from cardiac surgery. AB - OBJECTIVE: To measure predefined clinical effects resulting from the use of epidural anesthesia and analgesia during and after cardiac surgery. DESIGN: Prospective, randomized, nonblinded clinical trial. SETTING: Single academic medical center. PARTICIPANTS: Sixty patients scheduled for elective cardiac surgery with cardiopulmonary bypass. INTERVENTIONS: Sixty participants were randomly assigned to 1 of 2 study groups: (1) A control group received general anesthesia during surgery and intravenous opiate analgesia after surgery. (2) A treatment group received thoracic epidural anesthesia combined with general anesthesia during surgery and epidural analgesia for the first 24 postoperative hours. MEASUREMENTS AND MAIN RESULTS: Primary study measurements were planned to evaluate recovery from surgery and included time to tracheal extubation, duration of postoperative intensive care unit stay, duration of postoperative hospitalization, pain control, urinary free cortisol, cardiopulmonary complication rate, and total hospital charges. No statistically significant differences between the 2 study groups were found in these main measurements. CONCLUSIONS: The clinical course of elective cardiac surgical patients who receive epidural anesthesia during surgery and epidural analgesia after surgery is comparable to that of patients managed with general anesthesia alone during surgery followed by parenteral opiate analgesia after surgery. PMID- 11854873 TI - To ventilate or not after minimally invasive direct coronary artery bypass surgery: the role of epidural anesthesia. AB - OBJECTIVE: To evaluate the effect of immediate postoperative extubation and postoperative ventilation after minimally invasive direct coronary artery bypass (MIDCAB) surgery and to assess the role of epidural anesthesia. DESIGN: Randomized prospective study. SETTING: University hospital, single institution. PARTICIPANTS: Patients (n = 90) scheduled for elective MIDCAB surgery. INTERVENTIONS: Patients were divided into 3 groups: 30 patients had general anesthesia and were extubated immediately after surgery (extubated group), 30 patients had a thoracic epidural and general anesthesia and were extubated immediately after surgery (epidural group), and 30 patients had general anesthesia and were ventilated after surgery (intubated group). MEASUREMENTS AND MAIN RESULTS: With a similar cardiac index and less vasoactive medication, mean arterial blood pressure (77 plus minus 8 mmHg [mean plus minus SD]) and heart rate (76 plus minus 10 beats/min) in the epidural group were lower on the first postoperative day than in the intubated group (83 plus minus 10 mmHg and 81 plus minus 13 beats/min) and the extubated group (86 plus minus 10 mmHg and 83 plus minus 13) (p = 0.01 and p = 0.09). Oxygenation on the first postoperative day was better in the epidural group than in the intubated group (14.8 plus minus 3.8 kPa v 12.6 plus minus 3.2 kPa; p = 0.05). The epidural group and the extubated group had a transient respiratory acidosis postoperatively. Pain score in the epidural group was lower on the first postoperative day than in the extubated group with general anesthesia (3.0 plus minus 1.6 visual analog scale v 4.6 plus minus 1.8 visual analog scale; p = 0.01). Hospital stay was shorter in the epidural group than in the ventilated group (5.9 plus minus 2.4 days v 8.1 plus minus 5.3 days; p = 0.05) CONCLUSION: Immediate postoperative extubation in patients with thoracic epidural anesthesia and supplemental general anesthesia provides the most favorable clinical circumstances after MIDCAB surgery. PMID- 11854874 TI - Postoperative pulmonary function in coronary artery bypass graft surgery patients undergoing early tracheal extubation: a comparison between short-term mechanical ventilation and early extubation. AB - OBJECTIVE: To evaluate the effect of a short period of mechanical ventilation (3 hours) versus immediate extubation (within 1 hour of surgery) on pulmonary function, gas exchange, and pulmonary complications after coronary artery bypass graft (CABG) surgery. DESIGN: Prospective randomized study. SETTING: University teaching hospital. PARTICIPANTS: Thirty-five patients undergoing CABG surgery. INTERVENTIONS: Patients were randomized into 2 groups. Patients in group I were extubated as soon as possible after surgery. Patients in group II were ventilated for a minimum of 3 hours after surgery. Patients in both groups were extubated only after achieving predetermined extubation criteria. Patients who did not meet the criteria for extubation within the predetermined set time limit (90 minutes in group I and 6 hours in group II) were withdrawn from the study. Pulmonary function tests (vital capacity, forced expiratory volume in 1 second, total lung capacity, functional residual capacity), arterial blood gases, and chest radiographs were done preoperatively and postoperatively. Pulmonary complications were recorded. MEASUREMENTS AND MAIN RESULTS: Demographic data were similar between groups. The mean time to extubation in group I was 45.7 plus minus 27.6 minutes and in group II was 201.4 plus minus 21 minutes (p < 0.01). Two patients in group I and 1 patient in group II did not meet the extubation criteria within the predetermined set time limit and were excluded from the study. In both groups, there was a significant decline in pulmonary function but no differences between groups at 24 or 72 hours after surgery. There were no differences between groups in blood gases, atelectasis scores, or pulmonary complications. CONCLUSION: The data suggest that extending mechanical ventilation after CABG surgery does not affect pulmonary function. Provided that routine extubation criteria are met, patients can be safely extubated early (within 1 hour) after major cardiac surgery without concerns of further pulmonary derangement. PMID- 11854875 TI - Oxygen consumption after hypothermic cardiopulmonary bypass: the effect of continuing a propofol infusion postoperatively. AB - OBJECTIVE: To evaluate the effect of a fixed rate of infusion of propofol on total body oxygen consumption during the postoperative rewarming phase after cardiopulmonary bypass. DESIGN: Prospective, randomized, controlled study. SETTING: Cardiac intensive care unit, university hospital. PARTICIPANTS: Twenty four male and female patients undergoing elective first-time coronary artery bypass graft surgery. INTERVENTIONS: Total body oxygen consumption was measured using a pulmonary artery catheter and thermodilution during postoperative rewarming. Twelve patients had propofol infused at 2 mg/kg/h for 4 hours or until rewarmed. MEASUREMENTS AND MAIN RESULTS: Total body oxygen consumption was reduced in the propofol group compared with the control group. Oxygen consumption was a median of 30.0 mL/min/m(2) less in the patients receiving propofol (p = 0.01). One patient receiving propofol shivered compared with 4 in the control group (p = 0.14). CONCLUSION: Administration of propofol during postoperative rewarming reduces total body oxygen consumption and may reduce shivering. PMID- 11854876 TI - Heparin-bonded cardiopulmonary bypass circuits reduce cognitive dysfunction. AB - OBJECTIVE: To determine the incidence of cerebral dysfunction in cardiac surgical patients exposed to heparin-bonded cardiopulmonary bypass (HB-CPB) versus nonheparin-bonded cardiopulmonary bypass (NH-CPB) circuits through neuropsychometric testing and to correlate these findings with markers of the systemic inflammatory response to CPB. DESIGN: Prospective, randomized, blinded clinical trial. SETTING: University hospital. PARTICIPANTS: Sixty-one patients undergoing elective cardiac surgery. INTERVENTIONS: A cohort of 61 patients scheduled for elective coronary artery bypass graft surgery were prospectively randomized to receive either HB-CPB or NH-CPB circuits during surgery. Patients were evaluated for cerebral injury using a battery of neuropsychometric tests at the following 3 time points: (1) before surgery as a baseline examination, (2) postoperative day 5, and (3) postoperative week 6. Blood samples were drawn to measure inflammatory markers at the following time points: (1) preincision, after induction of anesthesia, (2) 15 minutes after onset of CPB, (3) 30 minutes after CPB, (4) 6 hours postoperatively, and (5) 24 hours postoperatively. MEASUREMENTS AND MAIN RESULTS: Neuropsychometric performance was evaluated by group-rate and event-rate analyses. By group-rate analysis, patients undergoing surgery with HB CPB performed significantly better at 5 days after surgery on 2 neuropsychometric tests (trails A [p < 0.01] and finger tapping with the dominant hand [p < 0.01]) and at 6 weeks after surgery on one neuropsychometric test (trails A [p < 0.01]). By event-rate analysis, at 5 days, patients undergoing surgery with HB-CPB circuits had less cognitive dysfunction (p < 0.05) compared with patients undergoing surgery with NH-CPB circuits. Serum samples were analyzed to evaluate markers of complement activation (C3a), proinflammatory cytokines (tumor necrosis factor-alpha, interleukin-1 beta, and interleukin-6), and coagulation (thrombin antithrombin complex [TAT]) using the quantitative sandwich enzyme immunoassay technique. Although there were no significant differences in cytokine activation in either group, C3a was significantly higher in the NH-CPB group intraoperatively at 1 hour after CPB (p < 0.05), and TAT was higher in the HB-CPB group at 24 hours after surgery (p < 0.05). CONCLUSIONS: Patients undergoing cardiac surgery with CPB have less postoperative cognitive dysfunction during CPB when HB-CPB circuits are employed. Although there was a relationship, this finding did not correlate with decreased complement activation intraoperatively and activation of coagulation postoperatively. PMID- 11854877 TI - Hypertonic saline (7.5%) decreases perioperative weight gain following cardiac surgery. AB - OBJECTIVE: To compare the effects of 7.5% hypertonic saline (HS) and 0.9% normal saline (NS) on perioperative weight gain in cardiac surgical patients. DESIGN: Prospective, randomized study. SETTING: University teaching hospital. PARTICIPANTS: Patients (n = 72) scheduled for elective coronary artery bypass graft surgery. INTERVENTIONS: Patients were randomly assigned to receive either 7.5% HS (36 patients) or 0.9% NS (36 patients) as a single dose of 4 mL/kg over 30 minutes during the postoperative rewarming phase in the intensive care unit. MEASUREMENTS AND MAIN RESULTS: Weight gain until the first postoperative morning was significantly greater in the NS group than in the HS group (1.9 plus minus 1.4 kg, median, 2.1 kg; 0.8 plus minus 1.5 kg, median, 0.8 kg; p = 0.005). One hour diuresis after the fluid infusion was significantly greater in the HS group compared with the NS group (501 plus minus 282 mL and 237 plus minus 173 mL; p < 0.001). In the linear regression model, the 2 most important factors affecting the perioperative weight gain were the volume of fluid infused postoperatively in the intensive care unit (4,098 plus minus 916 mL in the HS group and 4,589 plus minus 1,344 mL in the NS group) and the total diuresis after surgery (3,351 plus minus 1,035 mL in the HS group and 2,942 plus minus 846 mL in the NS group). CONCLUSIONS: HS had an intense diuretic effect, which reduced intraoperative fluid retention. This effect was confirmed by the lesser increase in body weight measured on the first postoperative morning. PMID- 11854878 TI - Hyperprocalcitonemia is related to noninfectious postoperative severe systemic inflammatory response syndrome associated with cardiovascular dysfunction after coronary artery bypass graft surgery. AB - OBJECTIVE: To investigate the role of 3 inflammatory parameters as early markers of severe systemic inflammatory response syndrome (SIRS) induced by coronary artery bypass graft surgery. DESIGN: Prospective study. SETTING: University hospital. PARTICIPANTS: Patients (n = 63) undergoing elective coronary artery bypass graft surgery with cardiopulmonary bypass. MEASUREMENTS AND MAIN RESULTS: The American College of Chest Physicians/Society of Critical Care Medicine classification was used to diagnose SIRS. Organ system failures were used to define severe SIRS. Serum concentrations of the inflammatory parameters (procalcitonin [PCT], C-reactive protein, leukocyte count) were determined before, during, and after surgery. SIRS occurred in 30 (47%) patients after surgery. Seven patients (11%) showed SIRS with greater-than-or-equal1 organ dysfunction (severe SIRS), whereas patients without SIRS had no organ dysfunction. Significantly higher serum levels of PCT were found in patients with severe SIRS from the 6th postoperative hour until the 3rd postoperative day with a peak level of 10.7 plus minus 13.2 ng/mL. No significant difference was detected between serum PCT of patients with SIRS but without any organ dysfunction and patients without SIRS. PCT levels of these patients remained lower than 1.7 ng/mL. Compared with PCT, plasma concentrations of C-reactive protein peaked later on the 2nd postoperative day and were not able to confirm the severity of SIRS. Leukocyte counts were not significantly modified. CONCLUSIONS: PCT seems to be an appropriate marker to identify the early development of noninfectious postoperative severe SIRS after coronary artery bypass graft surgery with cardiopulmonary bypass. PMID- 11854879 TI - The effect of preoperative aspirin-free interval on red blood cell transfusion requirements in cardiac surgical patients. AB - OBJECTIVE: To compare red blood cell transfusion in first-time coronary artery surgery patients who stopped taking aspirin < or = 2 days, 3 to 7 days, or >7 days preoperatively. DESIGN: Observational study. SETTING: University-affiliated teaching hospital. PARTICIPANTS: Adult patients (n = 797) undergoing first-time coronary artery surgery on cardiopulmonary bypass who were not receiving other anticoagulant or antiplatelet drugs before surgery. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Patients were divided into 4 groups based on days since last ingestion of aspirin. Blood products transfused in the groups were (aspirin < or =2 days) (n = 140) 2.2 +/- 4 U of red cell concentrate (RCC) (mean +/- SD), 1.4 +/- 3 U of fresh frozen plasma (FFP), and 2.7 +/- 6 U of platelets; (aspirin 3 to 5 days) (n = 255), 1.5 +/- 2 U of RCC, 0.8 +/- 2 U of FFP, and 1.6 +/- 4 U of platelets; (aspirin 6 to 7 days) (n = 215), 1.6 +/- 3 U of RCC, 0.9 +/ 3 U of FFP, and 1.5 +/- 3 U of platelets; and (aspirin >7 days) (n = 187), 1.3 +/- 2 U of RCC; 0.6 +/- 2 U of FFP, and 0.9 +/- 2 U of platelets. CONCLUSION: Patients who stop taking aspirin < or =2 s preoperatively have increased allogenic red blood cell transfusion requirements perioperatively. Patients who stop taking aspirin 3 to 7 days preoperatively have little or no increased requirement for allogenic red blood cell transfusion. PMID- 11854880 TI - Tissue factor pathway inhibitor antigen and activity in 96 patients receiving heparin for cardiopulmonary bypass. AB - OBJECTIVE: To identify patients with poor tissue factor pathway inhibitor (TFPI) response to heparin and observe any association with increased risk of excessive coagulation activation, morbidity, or mortality. DESIGN: Prospective, observational cohort study. SETTING: University hospital. PARTICIPANTS: Patients (n = 96) undergoing cardiopulmonary bypass for various types of surgery. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: TFPI antigen and activity were determined in patients before and after heparin administration, before cardiopulmonary bypass for cardiac surgery. The clinical progress of each patient was recorded. Median levels of TFPI activity were 0.98 U/mL (interquartile range, 0.83 to 1.14 U/mL) preheparin and 2.34 U/mL (2.18 to 2.54 U/mL) postheparin (p < 0.0001), representing a median 2.3-(2.1- to 2.8-) fold increase. Median TFPI antigen levels were 92.4 ng/mL (73.0 to 119.5 ng/mL) preheparin and 422.9 ng/mL (398.7 to 501.6 ng/mL) postheparin (p < 0.0001), representing a median 4.6-fold (3.6- to 6.2-fold) increase. Two patients had low (<300 ng/mL) postheparin levels of TFPI antigen that were not associated with low functional TFPI or adverse clinical outcome. Fourteen patients showed a low ratio of increased functional TFPI postheparin; all had a ratio of TFPI antigen increase of at least 3-fold. CONCLUSION: The TFPI response to heparin is heterogenous. Two nonresponders were identified, with low postheparin levels of TFPI antigen, who did not suffer adverse clinical outcomes. PMID- 11854881 TI - Milrinone increases middle cerebral artery blood flow velocity after cardiopulmonary bypass. AB - OBJECTIVE: To evaluate the effects of milrinone on middle cerebral artery blood flow velocity (Vmca) and pulsatility index (PI) during normocapnia and hyperventilation in adults after cardiopulmonary bypass (CPB). DESIGN: A prospective study. SETTING: University-affiliated hospital and Veterans Affairs Medical Center. PARTICIPANTS: Twenty-five adults with left ventricular ejection fraction >40% undergoing coronary artery bypass graft surgery. INTERVENTIONS: After separation from CPB, using transcranial Doppler ultrasonography, peak and mean Vmca and PI were recorded before and after the administration of 50 microg/kg of milrinone under normocapnia and with hyperventilation. MEASUREMENTS AND MAIN RESULTS: Heart rate, arterial blood pressure, central venous pressure, and cardiac output were documented after each study period. Compared with baseline, milrinone increased peak Vmca by 20%, increased mean Vmca by 19%, and decreased PI by 16% (p < 0.001). Before the administration of milrinone, hyperventilation decreased peak Vmca by 20%, decreased mean Vmca by 26%, and increased PI by 24% (p < 0.01). After milrinone administration, hyperventilation also decreased peak Vmca by 22%, decreased mean Vmca by 21%, and increased PI by 19% (p < 0.01). Milrinone increased cardiac index and decreased mean arterial pressure and systemic vascular resistance (p < 0.05); however, heart rate and central venous pressure remained unchanged. CONCLUSIONS: The administration of milrinone increases cerebral blood flow after CPB most likely as a result of cerebral vasodilation. The response to hyperventilation seems to be partially preserved. PMID- 11854882 TI - High-volume ultrafiltration with extracellular fluid replacement for the management of dialysis patients during cardiopulmonary bypass. PMID- 11854883 TI - Acidosis following aortic cross-clamping: is it the acid or carbon dioxide? PMID- 11854884 TI - Heparin resistance and Marfan syndrome: is there any correlation? PMID- 11854885 TI - Unusual intraoperative impact of transesophageal echocardiography during the correction of a persistent pseudoaneurysm perfused from the left ventricular outflow tract. PMID- 11854886 TI - Cardiopulmonary bypass and the systemic inflammatory response: effects on drug action. PMID- 11854887 TI - Point-of-care coagulation monitoring for cardiovascular patients: past and present. PMID- 11854888 TI - Case 1---2002---a patient with severe peripartum cardiomyopathy and persistent ventricular fibrillation supported by a biventricular assist device. PMID- 11854889 TI - Pro: atrial arrhythmia prophylaxis is required for cardiac surgery. PMID- 11854890 TI - Con: atrial arrhythmia prophylaxis is not required for cardiac surgery. PMID- 11854891 TI - Pulmonary edema following aortic valve replacement. PMID- 11854892 TI - Enlarged abnormal anatomic structure adjacent to the aortic valve. PMID- 11854893 TI - Updates in cardiopulmonary bypass. PMID- 11854894 TI - What is the optimal morphine dose to be administered intrathecally in postoperative analgesia of cardiac surgery? PMID- 11854895 TI - The use of alpha2-agonists during off-pump coronary artery bypass and the effects of hypothermia. PMID- 11854896 TI - Oesophageal surgery. PMID- 11854897 TI - Liver resection for cancer. PMID- 11854899 TI - Moving toward an understanding of the metastatic process in hepatocellular carcinoma. PMID- 11854898 TI - Pathogenesis of coeliac disease: implications for treatment. PMID- 11854900 TI - Apoptosis, proliferation and p53 gene expression of H. pylori associated gastric epithelial lesions. AB - AIM: To study the relationship between Helicobacter pylori (H. pylori) and gastric carcinoma and its possible pathogenesis by H. pylori. METHODS: DNEL technique and immunohistochemical technique were used to study the state of apoptosis, proliferation and p53 gene expression. A total of 100 gastric mucosal biopsy specimens, including 20 normal mucosa, 30 H. pylori-negative and 30 H. pylori-positive gastric precancerous lesions along with 20 gastric carcinomas were studied. RESULTS: There were several apoptotic cells in the superficial epithelium and a few proliferative cells within the neck of gastric glands, and no p53 protein expression in normal mucosa. In gastric carcinoma, there were few apoptotic cells, while there were a large number of proliferative cells, and expression of p53 protein significantly was increased. In the phase of metaplasia, the apoptotic index (AI, 4.36%+/-1.95%), proliferative index (PI, 19.11%+/-6.79%) and positivity of p53 expression (46.7%) in H. pylori-positive group were higher than those in normal mucosa (P<0.01). AI in H. pylori-positive group was higher than that in H. pylori-negative group (3.81%+/-1.76%), PI in H. pylori-positive group was higher than that in H. pylori-negative group (12.25%+/ 5.63%, P<0.01). In the phase of dysplasia, AI (2.31%+/-1.10%) in H. pylori positive group was lower (3.05%+/-1.29%) than that in H. pylori-negative group, but PI (33.89%+/-11.65%) was significantly higher (22.09+/-8018%, P<0.01). In phases of metaplasia, dysplasia and gastric cancer in the H. pylori-positive group, AIs had an evidently graduall decreasing trend (P<0.01), while PIs had an evidently gradual increasing trend (P<0.05 or P<0.01), and there was also a trend of gradual increase in the expression of p53 gene. CONCLUSION: In the course of the formation of gastric carcinoma, proliferation of gastric mucosa can be greatly increased by H. pylori, and H. pylori can induce apoptosis in the phase of metaplasia, but in the phase of dysplasia H. pylori can inhibit cellular apoptosis. And H. pylori infection can strengthen the expression of mutated p53 gene. PMID- 11854901 TI - Polymorphism of flagellin A gene in Helicobacter pylori. AB - AIM: To study the polymorphism of flagellin A genotype and its significance in Helicobacter pylori (H. pylori). METHODS: As the template, genome DNA was purified from six clinical isolates of H. pylori from outpatients, and the corresponding flagellin A fragments were amplified by polymerase chain reaction. All these products were sequenced. These sequences were compared with each other, and analyzed by software of FASTA program. RESULTS: Specific PCR products were amplified from all of these H. pylori isolates and no length divergence was found among them. Compared with each other, the highest ungapped identity is 99.10%, while the lowest is 94.65%. Using FASTA program, the alignments between query and library sequences derived from different H. pylori strains were higher than 90%. CONCLUSION: The nucleotide sequence of flagellin A in H. pylori is highly conservative with incident divergence. This information may be useful for gene diagnosis and further study on flagellar antigen phenotype. PMID- 11854903 TI - Cytochrome P450 2E1 genetic polymorphism and gastric cancer in Changle, Fujian Province. AB - AIM: Genetic polymorphism in enzymes of carcinogen metabolism has been found to have the influence on the susceptibility to cancer. Cytochrome P450 2E1 (CYP2E1) is considered to play an important role in the metabolic activation of procarcinogens such as N-nitrosoamines and low molecular weight organic compounds. The purpose of this study is to determine whether CYP450 2E1 polymorphisms are associated with risks of gastric cancer. METHODS: We conducted a population based case-control study in Changle county, Fujian Province, a high risk region of gastric cancer in China. Ninety-one incident gastric cancer patients and ninety-four healthy controls were included in our study. Datas including demographic characteristics, diet intake, and alcohol and tobacco consumption of individuals in our study were completed by a standardized questionnaire.PCR-RFLP revealed three genotypes:heterozygote (C1/C2) and two homozygotes (C1/C1 and C2/C2) in CYP2E1. RESULTS: The frequency of variant genotypes (C1/C2 and C2/C2) in gastric cancer cases and controls was 36.3% and 24.5%, respectively. The rare homozygous C2/C2 genotype was found in 6 individuals in gastric cancer group(6.6%), whereas there was only one in the control group (1.1%). However, there was no statistically significant difference between the two groups (two-tailed Fisher's exact test P=0.066). Individuals in gastric cancer group were more likely to carry genotype C1/C2 (odds ratio, OR=1.50) and C2/C2 (OR=7.34) than individuals in control group (chi(2) =4.597, for trend P=0.032). The frequencies of genotypes with the C2 allele (C1/C2 and C2/C2 genotypes) were compared with those of genotypes without C2 allele (C1/C1 genotype) among individuals in gastric cancer group and control group according to the pattern of gastric cancer risk factors. The results show that individuals who exposed to these gastric cancer risk factors and carry the C2 allele seemed to have a higher risk of developing gastric cancer. CONCLUSION: Polymorphism of CYP2E1 gene may have some effect in the development of gastric cancer in Changle county, Fujian Province. PMID- 11854902 TI - Human papillomavirus 16 E6 is associated with the nuclear matrix of esophageal carcinoma cells. AB - AIM: To explore the etiologic role of HPV infection in esophageal carcinoma, and the association of HPV-16 E6 with the nuclear matrix of carcinoma cells. METHODS: Two esophageal carcinoma cell lines,EC/CUHK1 and EC/CUHK2, were tested for HPV-16 E6 subgenetic fragment by polymerase chain reaction amplification of virus DNA associated nuclear matrix. RT-PCR and immunocytochemistry were also used to visualize the expression of E6 subgene in the cells. RESULTS: The HPV-16 E6 subgenetic fragment was found to be present in nuclear matrix-associated DNA, E6 oncoprotein localized in the nucleus where it is tightly associated with nuclear matrix after sequential extraction in EC/CUHK2 cells. It was not detected, however, in EC/CUHK1 cells. CONCLUSION: The interaction between HPV-16 E6 and nuclear matrix may contribute to the virus induced carcinogenesis in esophageal carcinoma. PMID- 11854904 TI - Growth inhibition and apoptosis induction of Sulindac on Human gastric cancer cells. AB - AIM: To evaluate the effects of sulindac in inducing growth inhibition and apoptosis of human gastric cancer cells in comparison with human hepatocellular carcinoma (HCC) cells. METHODS: The human gastric cancer cell lines MKN45 and MKN28 and human hepatocellular carcinoma cell lines HepG(2) and SMMC7721 were used for the study. Anti-proliferative effect was measured by MTT assay, and apoptosis was determined by Hoechst-33258 staining, electronography and DNA fragmentation. The protein of cyclooxygenase-2 (COX-2) and Bcl-2 were detected by Western dot blotting. RESULTS: Sulindac could initiate growth inhibition and apoptosis of MKN45, MKN28, HepG(2) and SMMC7721 cells in a dose-and time dependent manner. Growth inhibitory activity and apoptosis were more sensitive in HepG(2) cells than in SMMC7721 cells, MKN45 and MKN28 cells. After 24 hours incubation with sulindac at 2mmol x L(-1) and 4mmol x L(-1), the level of COX-2 and Bcl-2 protein were lowered in MKN45, SMMC7721 and HepG(2) cells but not in MKN28 cells. CONCLUSION: Sulindac could inhibit the growth of gastric cancer cells and HCC cells effectively in vitro by apoptosis induction, which was associated with regression of COX-2 and Bcl-2 expression. The growth inhibition and apoptosis of HCC cells were greater than that of human gastric cancer cells. The different effects of apoptosis in gastric cancer cells may be related to the differentiation of the cells. PMID- 11854905 TI - Association of H. pylori infection with gastric carcinoma: a Meta analysis. AB - AIM: To follow the principles of evidence based medicine to reach the integrated results of these studies. METHODS: Twenty-one papers of case-control studies were selected, including 11 on gastric cancer,7 on precancerous lesion of stomach and 3 on lymphoma of stomach. Meta analysis was used to sum up the odds ratios (OR) of these studies. RESULTS: H. pylori vs gastric cancer (intestinal and diffuse type): the odds ratio from the fixed effect model is 3.0016 (95% CI: 2.4197 3.7234, P<0.001). H. pylori vs precancerous lesion of stomach: a random effect model was used to calculate the summary odds ratio and its value is 2.5635 (95% CI: 1.8477-3.5566, P<0.01). H. pylori vs lymphoma of stomach: though the quantity of literature is too small to make Meta analysis, the data of these 3 studies show that lymphoma of stomach is highly associated with H. pylori infections. CONCLUSION: Since it had been revealed that H. pylori infection pre-exists in gastric carcinoma and precancerous lesions, the results of Meta analysis present a strong evidence to support the conclusion that H. pylori infection is a risk factor for gastric carcinoma. PMID- 11854906 TI - Clinicopathological and molecular genetic analysis of 4 typical Chinese HNPCC families. AB - AIM: To study the clinicopathological and molecular genetic characteristics of typical Chinese hereditary nonpolyposis cotorectal cancer (HNPCC) families. METHODS: Four typical Chinese HNPCC families were analyzed using microdissection, microsatellite instability analysis, immunostaining of hMSH2 and hMLH1 proteins and direct DNA sequencing of hMSH2 and hMLH1 genes. RESULTS: All five tumor tissues of 4 probands from the 4 typical Chinese HNPCC families showed microsatellite instability at more than two loci (MSI-H or RER+ phenotype). Three out of the 4 cases lost hMSH2 protein expression and the other case showed no hMLH1 protein expression. Three pathological germline mutations (2 in hMSH2 and 1 in hMLH1), which had not been reported previously, were identified. The same mutations were also found in other affected members of two HNPCC families,respectively. CONCLUSION: Typical Chinese HNPCC families showed relatively frequent germline mutation of mismatch repair genes. High-level microsatellite instability and loss of expression of mismatch repair genes correlated closely with germline mutation of mismatch repair genes. Microsatellite instability analysis and immunostaining of mismatch repair gene might serve as effective screening methods before direct DNA sequencing. It is necessary to establish clinical criteria and molecular diagnostic strategies more suitable for Chinese HNPCC families. PMID- 11854907 TI - Construction and selection of the natural immune Fab antibody phage display library from patients with colorectal cancer. AB - AIM: To construct the natural immune Fab antibody phage display libraries of colorectal cancer and to select antibodies related with colorectal cancer. METHODS: Extract total RNA from tissue of local cancer metastasis lymph nodes of patients with colorectal cancer. RT-PCR was used to amplify the heavy chain Fd and light chain kappa and the amplification products were inserted successively into the vector pComb3 to construct the human libraries of Fab antibodies. They were then panned by phage display technology. By means of Dot immunoblotting and ELISA, the libraries were identified and the Fab phage antibodies binding with antigens of colorectal cancer were selected. RESULTS: The amplified fragments of Fd and kappa gained by RT-PCR were about 650 bp. Fd and kappa PCR products were subsequently inserted into the vector pComb3, resulting in a recombination rate of 40% and the volume of Fab phage display library reached 1.48 x 10(6). The libraries were enriched about 120-fold by 3 cycles of adsorption-elution multiplication (panning). Dot immunoblotting showed Fab expressions on the phage libraries and ELISA showed 5 clones of Fab phage antibodies which had binding activities with antigens of colorectal cancer. CONCLUSION: The natural immune Fab antibody phage display libraries of colorectal cancer were constructed. They could be used to select the relative antibodies of colorectal cancer. PMID- 11854908 TI - SF/HGF-c-Met autocrine and paracrine promote metastasis of hepatocellular carcinoma. AB - AIM: To explore the role of SF/HGF-Met autocrine and paracrine in metastasis of hepatocellular carcinoma (HCC). METHODS: SF/HGF and c-met transcription and protein expression in HCC were examined by RT-PCR and Western Blot in 4 HCC cell lines, including HepG2, Hep3B, SMMC7721 and MHCC-1, the last cell line had a higher potential of metastasis. sf/hgf cDNA was transfected by the method of Lipofectin into SMMC7721. SF/HGF and c-met antibody were used to stimulate and block SF/HGF-c-met signal transduction. Cell morphology, mobility, and proliferation were respectively compared by microscopic observation, wound healing assay and cell growth curve. RESULTS: HCC malignancy appeared to be relative to its met-SF/HGF expression. In MHCC-1, c-met expression was much stronger than that in other cell lines with lower potential of metastasis and only SF/HGF autocrine existed in MHCC-1. After sf/hgf cDNA transfection or conditioned medium of MHCC-1 stimulation, SMMC7721 changed into elongated morphology, and the abilities of proliferation (P < 0.05) and mobility increased. Such bio-activity could be blocked by c-met antibody (P < 0.05). CONCLUSION: The system of SF/HGF-c-met autocrine and paracrine played an important role in development and metastasis potential of HCC. Inhibition of SF/HGF-c-met signal transduction system may reduce the growth and metastasis of HCC. PMID- 11854909 TI - Expression of liver cancer associated gene HCCA3. AB - AIM: To study and clone a novel liver cancer related gene, and to explore the molecular basis of liver cancer genesis. METHODS: Using mRNA differential display polymerase chain reaction (DDPCR), we investigated the difference of mRNA in human hepatocellular carcinoma (HCC) and paired surrounding liver tissues, and got a gene probe. By screening a human placenta cDNA library and genomic homologous extend, we obtained a full-length cDNA named HCCA3. We analyzed the expression of this novel gene in 42 pairs of HCC and the surrounding liver tissues, and distribution in human normal tissues by means of Northern blot assay. RESULTS: A full-length cDNA of liver cancer associated gene HCCA3 has been submitted to the GeneBank nucleotide sequence databases (Accession No. AF276707). The positive expression rate of this gene was 78.6% (33/42) in HCC tissues, and the clinical pathological data showed that the HCCA3 was closely associated with the invasion of tumor capsule (P=0.023) and adjacant small metastasis satellite nodules lesions (P=0.041). The HCCA3 was widely distributed in the human normal tissues, which was intensively expressed in lungs, brain and colon tissues, while lowly expressed in the liver tissues. CONCLUSION: A novel full-length cDNA was cloned and differentiated, which was highly expressed in liver cancer tissues. The high expression was closely related to the tumor invasiveness and metastasis,that may be the late heredited change in HCC genesis. PMID- 11854910 TI - TECA hybrid artificial liver support system in treatment of acute liver failure. AB - AIM: To assess the efficacy and safety of TECA type hybrid artificial liver support system (TECA-HALSS) in providing liver function of detoxification, metabolism and physiology by treating the patients with acute liver failure (ALF). METHODS: The porcine liver cells (1-2) x 10(10) were separated from the Chinese small swine and cultured in the bioreactor of TECA-BALSS at 37.0 degrees C and circulated through the outer space of the hollow fiber tubes in BALSS. The six liver failure patients with various degree of hepatic coma were treated by TECA-HALSS and with conventional medicines. The venous plasma of the patients was separated by a plasma separator and treated by charcoal adsorbent or plasma exchange. The plasma circulated through the inner space of the hollow fiber tubes of BALSS and mixed with the patients' blood cells and flew back to their blood circulation. Some small molecular weight substances were exchanged between the plasma and porcine liver cells. Each treatment lasted 6.0-7.0 h. Physiological and biochemical parameters were measured before,during and after the treatment. RESULTS: The average of porcine liver cells was (1.0-3.0) x 10(10) obtained from each swine liver using our modified enzymatic digestion method. The survival rate of the cells was 85%-93% by trypan blue stain and AO/PI fluorescent stain. After cultured in TECA-BALSS bioreactor for 6 h, the survival rate of cells still remained 70%-85%. At the end of TECA-HALSS treatment, the levels of plasma NH(3), ALT, TB and DB were significantly decreased. The patients who were in the state of drowsiness or coma before the treatment improved their appetite significantly and regained consciousness, some patients resumed light physical work on a short period after the treatment. One to two days after the treatment, the ratio of PTA increased warkedly. During the treatment, the heart rates, blood pressure, respiration condition and serum electrolytes (K(+), Na(+) and Cl(-)) were stable without thrombosis and bleeding in all the six patients. CONCLUSION: TECA-HALSS treatment could be a rapid, safe and efficacious method to provide temporary liver support for patients with ALF. PMID- 11854911 TI - Chiral metabolism of propafenone in rat hepatic microsomes treated with two inducers. AB - AIM: To study the influence of inducers of drug metabolism enzyme, beta naphthoflavone (BNF) and dexamethasone (DEX), on the stereoselective metabolism of propafenone in the rat hepatic microsomes. METHODS: Phase I metabolism of propafenone was studied using the microsomes induced by BNF and DEX and the non induced microsome was used as the control. The enzymatic kinetics parameters of propafenone enantiomers were calculated by regress analysis of Eadie-Hofstee Plots. Propafenone enantiomer concentrations were assayed by a chiral HPLC. RESULTS: The metabolite of propafenone, N-desalkylpropafenone, was found after incubation of propafenone with the rat hepatic microsomes induced by BNF and DEX. In these two groups, the stereoselectivity favoring R(-) isomer was observed in metabolism at low substrate concentrations of racemic propafenone, but lost the stereoselectivity at high substrate concentrations. However, in control group, no stereoselectivity was observed. The enzyme kinetic parameters were: (1) K(m). CONTROL GROUP: R(-) 83+/-6, S(+) 94+/-7; BNF group: R(-) 105+/-6, S(+)128+/-14; DEX group: R(-) 86+/-11, S(+) 118+/-16; (2)V(max). CONTROL GROUP: R(-) 0.75+/ 0.16, S(+) 0.72+/-0.07; BNF group: R(-) 1.04+/-0.15, S(+)1.07+/-14; DEX group: R( ) 0.93+/-0.06, S(+) 1.04+/-0.09; (3)Cl(int). CONTROL GROUP: R(-) 8.9+/-1.1, S(+) 7.6+/-0.7; BNF group: R(-) 9.9+/-0.9, S(+)8.3+/-0.7; DEX group: R(-) 10.9+/-0.8, S(+) 8.9+/-0.9. The enantiomeric differences in K(m) and Cl(int) were both significant, but not in V(max), in BNF and DEX group. Whereas enantiomeric differences in three parameters were all insignificant in control group. Furthermore, K(m) and V(max) were both significantly less than those in BNF or DEX group. In the rat liver microsome induced by DEX, nimodipine (NDP) decreased the stereoselectivity in propafenone metabolism at low substrate concentration. The inhibition of NDP on the metabolism of propafenone was stereoselective with R(-)-isomer being impaired more than S(+)-isomer. The inhibition constant (Ki) of S(+)- and R(-)-propafenone, calculated from Dixon plots, was 15.4 and 8.6 mg x L( 1), respectively. CONCLUSION: CYP1A subfamily(induced by BNF) and CYP3A4 (induced by DEX) have pronounced contribution to propafenone N-desalkylation which exhibited stereoselectivity depending on substrate concentration. The molecular base for this phenomenon is the stereoselectivity in affinity of substrate to the enzyme activity centers instead of at the catalyzing sites. PMID- 11854912 TI - Expression of lipopolysaccharide binding protein and its receptor CD14 in experimental alcoholic liver disease. AB - AIM: To evaluate the relationship between the expression of lipopolysaccharides (LPS) binding protein (LBP) and CD14 mRNA and the severity of liver injury in alcohol-fed rats. METHODS: Twenty Wistar rats were divided into two groups:ethanol-fed group (group E) and control group (group C). Group E was fed with ethanol(5-12 g x kg(-1) x d(-1)) and group C received dextrose instead of ethanol. Rats of the two groups were sacrificed at 4 weeks and 8 weeks. Levels of endotoxin and alanine transaminase (ALT) in blood were measured, and liver pathology was observed under light and electronic microscopy. Expressions of LBP and CD14 mRNA in liver tissues were determined by RT-PCR analysis. RESULTS: Plasma endotoxin levels were increased more significantly in group E(129+/-21) ng x L(-1) and (187+/-35) ng x L(-1) at 4 and 8 wk than in control rats(48+/-9) ng x L(-1) and (53+/-11) ng x L(-1), respectively (P<0.05). Mean values of plasma ALT levels were (1867+/-250) nkat x L(-1) and (2450+/-367) nkat x L(-1) in Group E. The values were increased more dramatically in ethanol-fed rats than in Group C after 4 and 8 weeks. In liver section from ethanol-fed rats, there were marked pathological changes (steatosis, cell infiltration and necrosis). In ethanol-fed rats, ethanol administration led to a significant increase in LBP and CD14 mRNA levels compared with the control group (P<0.05). CONCLUSION: Ethanol administration led to a significant increase in endotoxin levels in serum and LBP and CD14 mRNA expressions in liver tissues. The increase of LBP and CD14 mRNA expression might wake the liver more sensitive to endotoxin and liver injury. PMID- 11854913 TI - Cloning of UGT1A9 cDNA from liver tissues and its expression in CHL cells. AB - AIM: To clone the cDNA of UGT1A9 from a Chinese human liver and establish the Chinese hamster lung (CHL) cell line expressing human UGT1A9. METHODS: cDNA of UGT1 A9 was transcripted from mRNA by reverse transcriptase-ploymerase chain reaction, and was cloned into the pGEM-T vector which was amplified in the host bacteric E.Coli DH5(alpha). The inserted fragment, verified by DNA sequencing, was subcloned into the Hind III /Not I site of a mammalian expression vector pREP9 to construct the plasmid termed pREP9-UGT1A9. CHL cells were transfected with the resultant recombinants, pREP9-UGT1A9, and selected by G418 (400 mg x L( 1)) for one month. The surviving clone (CHL-UGT1A9) was harvested as a pool and sub-cultured in medium containing G418 to obtain samples forUGT1A9 assays. The enzyme activity of CHL-UGT1A9 towards propranolol in S9 protein of the cell was determined by HPLC. RESULTS: The sequence of the cDNA segment cloned, which was 1666 bp in length, was identical to that released by Gene Bank (GenBank accession number: AF056188) in coding region. The recombinant constructed, pREP9-UGT1A9, contains the entire coding region, along with 18 bp of the 5' and 55 bp of the 3' untranslated region of theUGT1A9 cDNA, respectively. The cell lines established expressed the protein of UGT1A9, and the enzyme activity towards propranolol in S9 protein was found to be 101+/- 24 pmol x min(-1) x mg(-1) protein (n=3), but was not detectable in parental CHL cells. CONCLUSION: The cDNA of UGT1A9 was successfully cloned from a Chinese human liver and transfected into CHL cells. The CHL-UGT1 A9 cell lines established efficiently expressed the protein ofUGT1A9 for the further enzyme study of drug glucuronidation. PMID- 11854914 TI - Copper transportion of WD protein in hepatocytes from Wilson disease patients in vitro. AB - AIM: To study the effect of copper transporting P-type ATPase in copper metabolism of hepatocyte and pathogenesis of Wilson disease (WD). METHODS: WD copper transporting properties in some organelles of the cultured hepatocytes were studied from WD patients and normal controls.These cultured hepatocytes were incubated in the media of copper 15 mg x L(-1) only, copper 15 mg x L(-1) with vincristine (agonist of P-type ATPase) 0.5mg x L(-1), or copper 15 mg x L(-1) with vanadate (antagonist of P-type ATPase) 18.39 mg x L(-1) separately. Microsome (endoplasmic reticulum and Golgi apparatus), lysosome, mitochondria, and cytosol were isolated by differential centrifugation. Copper contents in these organelles were measured with atomic absorption spectrophotometer, and the influence in copper transportion of these organelles by vanadate and vincristine were comparatively analyzed between WD patients and controls. WD copper transporting P-type ATPase was detected by SDS-PAGE in conjunction with Western blot in liver samples of WD patients and controls. RESULTS: The specific WD proteins (M(r)155,000 lanes) were expressed in human hepatocytes, including the control and WD patients. After incubation with medium containing copper for 2 h or 24 h, the microsome copper concentration in WD patients was obviously lower than that of controls, and the addition of vanadate or vincristine would change the copper transporting of microsomes obviously. When incubated with vincristine, levels of copper in microsome were significantly increased, while incubated with vanadate, the copper concentrations in microsome were obviously decreased. The results indicated that there were WD proteins, the copper transportion P-type ATPase in the microsome of hepatocytes. WD patients possessed abnormal copper transporting function of WD protein in the microsome, and the agonist might correct the defect of copper transportion by promoting the activity of copper transportion P-type ATPase. CONCLUSION: Copper transportion P-type ATPase plays an important role in hepatocytic copper metabolism. Dysfunction of hepatocytic WD protein copper transportion might be one of the most important factors for WD. PMID- 11854915 TI - Distribution of nitric oxide synthase in stomach myenteric plexus of rats. AB - AIM: To study the distribution of nitric oxide synthase (NOS) in rat stomach myenteric plexus. METHODS: The distribution of NOS in gastric wall was studied in quantity and location by the NADPH-diaphorase (NDP) histochemical staining method and whole mount preparation technique. RESULTS: NOS was distributed in whole stomach wall, most of them were located in myenteric plexus, and distributed in submucosal plexus.The shape of NOS positive neurons was basically similar, most of them being round and oval in shape. But their density, size and staining intensity varied greatly in the different parts of stomach. The density was 62+/ 38 cells mm(2) (antrum), 43+/-32 cells/mm(2) (body), and 32+/-28 cells mm(2) (fundus), respectively. The size and staining intensity of NOS positive neurons in the fundus were basically the same, the neurons being large and dark stained, while they were obviously different in antrum. In the body of the stomach, the NOS positive neurons were in an intermediate state from fundus to antrum. There were some beadlike structures which were strung together by NOS positive varicosities in nerve fibers, some were closely adherent to the outer walls of blood vessels. CONCLUSION: Nitric oxide might be involved in the modulation of motility, secretion and blood circulation of the stomach, and the significant difference of NOS positive neurons in different parts of stomach myenteric plexus may be related to the physiologic function of stomach. PMID- 11854916 TI - Taxotere resistance in SUIT Taxotere resistance in pancreatic carcinoma cell line SUIT 2 and its sublines. AB - AIM: To investigate the specific mechanisms of intrinsic and acquired resistance to taxotere (TXT) in pancreatic adenocarcinoma (PAC). METHODS: MTT assay was used to detect the sensitivity of PAC cell line SUIT-2 and its sublines (S-007, S-013, S-020, S-028 and TXT selected SUIT-2 cell line, S2/TXT) to TXT. Mdr1 (P-gp), multidrug resistance associated protein (MRP), lung resistance protein (LRP) and beta-tubulin isotype gene expressions were detected by RT-PCR. The functionality of P-gp and MRP was tested using their specific blocker verapamil (Ver) and indomethacin (IMC), respectively. The transporter activity of P-gp was also confirmed by Rhodamine 123 accumulation assay. RESULTS: S-020 and S2/TXT were found to be significantly resistant to TXT(19 and 9.5-fold to their parental cell line SUIT-2, respectively). RT-PCR demonstrated strong expression of Mdr1 in these two cell lines, but weaker expression or no expression in other cells lines. MRP and LRP expressions were found in most of these cell lines. The TXT resistance in S2-020 and S2/TXT could be reversed almost completely by Ver, but not by IMC. Flow cytometry showed that Ver increased the accumulation of Rhodamine-123 in these two cell lines. Compared with S-020 and SUIT-2, the levels of beta-tubulin isotype II, III expressions in S-2/TXT were increased remarkably. CONCLUSION: The both intrinsic and acquired TXT-related drug resistance in these PAC cell lines is mainly mediated by P-gp, but had no relationship to MRP and LRP expressions. The increases of beta-tubulin isotype II, III might be collateral changes that occur when the SUIT-2 cells are treated with TXT. PMID- 11854917 TI - Fas counterattack in cholangiocarcinoma: a mechanism for immune evasion in human hilar cholangiocarcinomas. AB - AIM: To investigate FasL expression in hilar cholangiocarcinoma tissues and cultured cholangiocarcinoma cells, and to assess its ability to induce apoptosis. METHODS: We studied the expression of FasL by human hilar cholangiocaroinomas tissues by immunohistochemistry, and the QBC939 cholangiocarcinoma cell line by RT-PCR, immunohistochemistry, and Western Blot. TUNEL and flow cytometry were used to detect apoptotic cells. RESULTS: Prevalent expression of FasL was detected in 39 resected hilar cholangiocarcinoma tissues. TUNEL staining disclosed a high level of cell death among lymphocytes infiltrating FasL positive areas of tumor. FasL mRNA and protein expressions in cholangiocarcinoma cells could induce Jurkat cells. CONCLUSION: Hilar cholangiocarcinomas may elude immunological surveillance by inducing, via Fas/FasL system, the apoptosis of activated lymphocytes. PMID- 11854919 TI - Preliminary research on myosin light chain kinase in rabbit liver. AB - AIM: To study preliminarily the properties of myosin light chain kinase (MLCK) in rabbit liver. METHODS: The expression of MLCK was detected by reverse transcription-polymerase chain reaction(RT-PCR); the MLCK was obtained from rabbit liver, and its activity was analyzed by gamma-(32)P incorporation technique to detect the phosphorylation of myosin light chain. RESULTS: MLCK was expressed in rabbit liver, and the activity of the enzyme was similar to rabbit smooth muscle MLCK, and calmodulin-dependent. When the concentration was 0.65 mg x L(-1), the activity was at the highest level. CONCLUSION: MLCK expressed in rabbit liver may catalyze the phosphorylation of myosin light chain, which may play important roles in the regulation of hepatic cell functions. PMID- 11854918 TI - Relationship between genotype and phenotype of flagellin C in Salmonella. AB - AIM: To discover the relationship between the genotype and antigen serotype of flagellin C among Salmonella strains. METHODS: Fragment of Salmonella flagellin C in plasmid pLS408 was cloned, sequenced and compared with the corresponding sequence in other strains. Salmonella strains including two typhi strains, one paratyphoid strain, one enteritidis and one typhimurium strain were isolated from outpatients. Genome DNA was purified respectively from these clinical isolates, then the corresponding flagellin C fragment was amplified by polymerase chain reaction,and the amplification products were analyzed by agarose gel electrophoresis. RESULTS: The cloned fragment includes 582 nucleotides encoding the variable region and partial conservative region of Salmonella flagellin C in plasmid pLS408. With comparison to the corresponding sequences reported previously, there is only a little difference from other strains with the same flagellar serotype in both nucleotide and amino acid level. Specific PCR products were amplified in Salmonella strains with flagellar serotype H-1-d including S. muenchen, typhi and typhimurium, but not in S. paratyphoid C or S. enteritidis strains. CONCLUSION: In this experiment, the specificity of nucleotide sequence could be found in flagellin C central variable regions as it exists in flagellar serotypes in Salmonella. It may be helpful to developing a rapid, sensitive, accurate and PCR-based method to detect Salmonella strains with serotype H-1-d. PMID- 11854920 TI - Influence of BOL on hyaluronic acid, laminin and hyperplasia in hepatofibrotic rats. AB - AIM: To study the anti-hepatofibrosis mechanism of Bie Jia Jian oral liquid (BOL). METHODS: The model was induced by subcutaneous injection of CCl(4). BOL was administered and the change of serum hyaluronic acid (HA) and laminin (LN) was observed and the degeneration of liver cells and the degree of fibre hyperplasia analyzed. Changes of ultra micro-structure in liver cells were observed in some samples. RESULTS: HA was reduced in both the groups with low and high dosage of BOL, which showed a remarkable difference as compared with that of the model group (low dosage group: 376.15 microg/L+/-35.48 microg/L vs 806.07 microg/L+/-98.49 microg/L P<0.05; high dosage group: 340.14 microg/L+/-30.18 microg/L vs 806.07 microg/L+/-98.49 microg/L P<0.05). The LN content of low and high dosage group of BOL was lower than that of model group (low dosage group: 71.99 microg/L+/-8.15 microg/L vs 133.94 microg/L+/-14.45 microg/L P <0.01; high dosage group: 71.68 microg/L+/-11.62 microg/L vs 133.94 microg/L+/-14.45 microg/L P<0.01) and colchicine group (low dosage group: 71.99 microg/L+/-8.15 microg/L vs 118.28 microg/L+/-16.13 microg/L P < 0.05; high dosage group: 71.68 microg/L+/ 11.62 microg/L vs 118.28 microg/L+/-16.13 microg/L P <0.05). Examined by Ridit, BOL could reduce the degeneration and necrosis of liver cells (chi(2)=11.99 P<0.05), the degree of fibre hyperplasia (chi(2)=13.24 P<0.05) and the pathological change of ultra micro-structure as well. CONCLUSION: The BOL has certain therapeutic effect on the experiment hepatofibrosis. Its mechanisms might include: protecting the function of liver cells, inhibiting excessive synthesis and secretion of extracellular matrix from hepatic stellate cells, relieving the capillarization of hepatic sinusoid, improving liver micro-circulation, and regulating immune function. PMID- 11854922 TI - Percutaneous transsplenic embolization of esophageal and gastrio-fundal varices in 18 patients. AB - AIM: Clinical application and potential complication of percutaneous transsplenic varices embolization (PTSVE) of esophageal or gastrio-fundal varices in patients with hepatocellular carcinoma (HCC) complicated with portal vein cancerous thrombosis (PVCT). METHODS: 18 patients with HCC complicated with PVCT and esophageal or gastrio-fundal varices who underwent PTSVE were collected. The rate of success, complication, mortality of the procedure and postoperative complication were recorded and analyzed. RESULTS: PTSVE were successfully performed in 16 of 18 cases, and the rate of success was 89%. After therapy erythrocyte counts decreased in all of the natunts. 5 of patients needed blood transfusion, 2 patients required surgical intervention because of and 11 patients with ascites were alleviated by diuresis. Among these 18 patients, the procedure related mortality was 11% (2/18), one died of acute hepatic failure on the forth day after procedure, another died of acute renal failure on the fifth day. The patients were follow up for 1-12 mon exceptone. 13 of them died of their tumors but none of them experienced variceal bleeding. CONCLUSION: PTSVE is a relatively safe and effective method to treat esophageal or gastrio-fundal varices in HCC patients with PVCT when percutaneous transhepatic varices embolization (PTHVE) of varices is impossible. PMID- 11854923 TI - Rapid donor liver procurement with only aortic perfusion. AB - AIM: To describe a rapid technique for procurement of donor liver with aortic perfusion only (APO). METHODS: Only the aorta is cannulated and perfused with chilled preservation solution. RESULTS: The quality of donor liver can ensure the grafted liver functions. CONCLUSION: The method of APO can simplify the operative procedure, compared with the dual cannulation. It also can minimize the danger of injuring vascular structures and involve less dissection. PMID- 11854921 TI - Clinical application of serial operations with preserving spleen. AB - AIM: To evaluate the clinical application of serial operations with preservation of spleen. METHODS: Serial operations with preserving spleen were performed on 211 cases in our hospital from 1980 to 2000. The patient's age ranged from 13 to 56 years, averaging 38 years. Diseases included splenic injury in 171 cases, portal hypertension in 9 cases, splenic cyst in 10 cases, and the lesion of pancreatic body and tail in 21 cases. RESULTS: All the cases were cured, and 129 patients were followed up from 3 months to 3 years with the leukocyte phagocytosis test, detection of immunoglubin, CT,(99m)Tc scanning and ultrasonography. The results were satisfactory. CONCLUSION: The operations with preserving spleen were safe, feasible, and worth of clinical application. PMID- 11854924 TI - SS-penogram: a new diagnostic test for erectile dysfunction. AB - The clinical reports on Sildenafil sulfate (Viagra) are mainly based on individual observations. However, there is a paucity of objective studies in the literature. In order to objectively examine the effect of Sildenafil, a SS (Sexual Stimulation)-Penogram that is a non-invasive, simple and physiologic method was developed using a radioisotope (RI). One hundred and four SS-penograms were performed on patients who had a documented erectile dysfunction (ED) lasting for more than 6 months. After an intravenous injection of 99mTc-RBC (15 mCi), the first penogram was taken immediately after sexual stimulation, which was done by 30 minutes of erotic videotape viewing. Forty minutes after administering 25 to 100 mg of Sildenafil, a second penogram was taken. The characteristics of each penogram were analyzed according to a previously reported method. The results were graded as follows; Type I(normal function; 5 min or more of peak erectile response with an induction period of 1 to 6 min), Type II-A (impossible function type; i.e., showing less than 2 times the basal radioactivity level), Type II-B (the unstable type; showing less than 5 min of peak erectile response), and Type II-C (the delayed type; which showed a delay of more than 15 min after the start of sexual stimulation). The patients were grouped according to their response after Sildenafil administration, and the effect of Sildenafil was assessed by comparing the radioactivity from between 7 to 22 minutes and the changes in the characteristics of the penogram. The mean age of the patients was 44.9 +/- 10.2 (23 - 68) years. In the first penogram, Type I was found in 12 patients, and Type II-A in 14, Type II-B in 73, Type II-C in 1 and a mixed (II-B + C) type was found in 4 patients. A second penogram after Sildenafil administration, showed Type I in 46 patients, and Type II-A in 10, Type II-B in 46 and a mixed type was found in 2 patients. The responses after Sildenafil were categorized as follows: 1) An excellent response group (consisting of 56 patients-53.9%); Those who showed greater than 50% increase in the RI area after Sildenafil treatment. 2) A good response group consisting of (23 patients-22.1%); i.e., those who showed a less than 50% but greater than a 20% increase in the RI area after Sildenafil administration. 3) A borderline group (consisting of 15 patients-14.4%); showing less than a 20% change in the RI area after Sildenafil treatment. 4) non-response group (consisting of 10 patients-9.6%). The therapeutic efficacy of Sildenafil, as determined by the SS-penograms, revealed that there was an augmentation in the erectile capabilities in 76% of men (79/104) but a non-response was observed in 9.6% (10/104). The efficacy of Sildenafil on the SS-penogram did not correlate with the patient's age (p=0.198). It is believed that the SS-penogram can be used to accurately evaluate the natural erectile status in sexual and pharmacological stimulation, and provides the most objective erectile response in any therapeutic trial. Consequently, the primary challenge for any erectile dysfunction remedy is to be able to demonstrate its efficacy. A further evaluation is warranted in the non-response group, which was not based on any severe organic dysfunction. PMID- 11854925 TI - Detection of Helicobacter pylori antigen in stool by enzyme immunoassay. AB - Invasive techniques for diagnosis of Helicobacter pylori (H. pylori) infection require an endoscopic examination which is expensive and inconvenient and may cause complications. Stool cultures for H. pylori or a direct detection of H. pylori antigen in stools by PCR are expensive, tedious, and have a low sensitivity. We recently used an enzyme immunoassay (EIA) to detect H. pylori antigen in stool specimens. A total of 41 patients were seen at Inha University Hospital, Inchon, Korea between September and October 1998. There were 26 men and 15 women who had an average age of 37.6 years which ranged from 5 to 71 years in the present study. All of these patients came to the hospital complaining of an upper abdominal discomfort and were subjected to endoscopy and biopsies. Fifteen had a gastric ulcer, 13 had a duodenal ulcer, 1 had an early gastric cancer, and there were 12 chronic gastritis patients as shown by endoscopy. The biopsy specimens were examined by histology, CLO test, and cultures and these results were used as gold standards. Stool specimens were tested for the H. pylori antigen by EIA. A dual wavelength cut-off of 0.100 that was recommended by the manufacturer gave a good performance (87.1% sensitivity, 100% specificity, 100% positive predictive value, 71.4% negative predictive value, and a 90.2% efficiency). But the adjusted cut-off value using the receiver operating characteristic curve improved the performance of the test (using the cut-off value of 0.024, the sensitivity, specificity, PPV, NPV, and efficiency were 100%, 90.0%, 96.9%, 100%, and 97.6% respectively). Re-evaluation of the cut-off value may be needed for Korean patients. This technique is non-invasive, rapid, easy-to use, and shows good performance characteristics for diagnosis of H. pylori infections. Therefore, this technique may be a substitute for gastric endoscopy especially in children and some patients who are unable to tolerate an endoscopic examination and it may be substituted for a serologic test in epidemiological research. PMID- 11854926 TI - Analysis of factors associated with the workers' health status using periodic health examination data by size of enterprises. AB - This study was conducted to determine whether the size of an enterprise can make a difference to workers' health and to examine those factors, which influence workers' health. The subjects of this study included 26,324 workers employed in manufacturing industry who received medical examinations at the Industrial Health Center at Yonsei University in 1995, 1996 and 1997. Medical examination data were used as a measure of health. Each enterprise was classified by size into three categories: Small scale enterprises (SSE) that employ no more than 50 regular employees; middle scale enterprise (MSE) that employ from 50 to 300 regular employees; and large scale enterprise (LSE) that employ more than 300 regular employees. Health conditions, according to the size of the enterprise, were determined by comparing prevalence rates, and direct standardization methods were used based upon the Korean population, as a standard population, in order to adjust for differences in population structure. 1. The findings of the medical examinations showed that, the prevalence rate in SSEs was higher than in MSEs or LSEs, but that this relationship was reversed after standardizing for sex and age. 2. Logistic regression analysis showed that, although subjects of advanced age, prolonged work history, and male gender had higher risks of disease, the size of the enterprise had no significant effect upon these risks. PMID- 11854927 TI - Monitoring of PETCO2 during high frequency jet ventilation for laryngomicrosurgery. AB - In general, PETCO2 is well correlated with PaCO2 during spontaneous and conventional mechanical ventilation in normal lungs. However, it is known that during high frequency jet ventilation, PETCO2 may underestimate PaCO2 because of inadequate washout of the anatomical dead space by a small tidal volume and the relatively slow response time of infrared CO2 analyzers. The validity of PETCO2 as a reflection of PaCO2 was assessed during HFJV in 40 patients undergoing laryngeal microsurgery. HFJV was applied through an injector inserted into the trachea 6 cm below the vocal cord. PETCO2 was obtained from a sampling line placed 2 cm below the injector. Both PETCO2 and PaCO2 were measured simultaneously after decreasing the frequency from 100 beats per minute to 15 beats per minute 10 and 20 minutes after the commencement of HFJV. There was a strong correlation (r = 0.955, P < 0.001) and a good correspondence between the mean PETCO2 and PaCO2 values with an average difference of 1.93 +/- 1.21 mmHg and a limit of agreement from -0.49 to 4.35 mmHg. It is suggested that the PETCO2 obtained following a decrease in the jet frequency during HFJV could closely reflect PaCO2. PMID- 11854928 TI - Synchrotron radiation imaging of internal structures in live animals. AB - Ionizing radiation has long been used in medicine since the discovery of X-rays. Diagnostic imaging using synchrotron radiation has been under investigation since Rubenstein et al. reported dual-energy iodine-K-edge subtraction coronary angiography. Recently, computed tomography (CT) and magnetic resonance imaging (MRI) have provided better quality results than conventional radiology, providing important information on human internal structures. However, such techniques are unable to detect fine micron sized structures for the early diagnosis of tumors, vascular diseases and other medical objectives. Third generation synchrotron X rays are well known for their superiority in coherence and energy tunability with respect to conventional X-rays. Consequently, new contrast mechanisms with a superior spatial resolution are becoming available. Here we present the extremely fine details of live animal internal structures using unmonochromatized synchrotron X-rays (white beam) and a simple detector system. Natural movements of the internal organs are also shown. The results indicate that this imaging technique can be applied to investigating microstructures and evaluating the function of the internal organs. Furthermore, this imaging system may be applied to humans as the next tool beyond CT and MRI. PMID- 11854929 TI - Morphometric evaluation of PGP9.5 and NCAM expressing nerve fibers in colonic muscle of patients with Hirschsprung's disease. AB - A quantitative assessment of the density of the protein gene product 9.5 (PGP9.5), the neural cell adhesion molecule (NCAM), and the low-affinity nerve growth factor receptor (NGFR) expressing nerve fibers in the circular muscle layer in the colon was carried out by morphometric analyses from 13 patients with Hirschsprung's disease (HD). The difference in the nerve fiber density between the ganglionic and aganglionic segments was compared by calculating the ratio of the sum of the areas occupied by positively stained nerve fibers per unit area of the muscle after immunohistochemical staining on paraffin embedded tissue sections using computer software. There was an obvious difference in the density of the PGP9.5 stained nerve fibers between the ganglionic (0.0380 +/- 0.0171) and aganglionic segments (0.0143 +/- 0.01661). The NCAM-positive nerve fibers were fewer in number than those of both the PGP9.5-positive fibers and NCAM-positive fibers, which were also markedly lower in number in the aganglionic segment (0.0066 +/- 0.0076) than in the ganglionic segment (0.0230 +/- 0.0195). Immunostaining for low-affinity NGFR revealed much fainter staining in the ganglionic and aganglionic segment without a statistically significant difference in their density. Considering the fact that PGP9.5 is a very sensitive marker for nerve fibers, the results of this study reaffirm the innervation failure of the proper muscle in HD. The decreased NCAM expression level in the aganglionic segment appears to be caused not by the selective down-regulation of NCAM expression among the nerve fibers but by a markedly reduced number of nerve fibers. PMID- 11854930 TI - Evaluation of CSF motion in syringomyelia with spatial modulation of magnetization (SPAMM). AB - CSF flow dynamics has been known to have a key role the pathogenesis of syringomyelia. Our purpose was to evaluate the CSF flow pattern in syringomyelia with spatial modulation of magnetization (SPAMM) that can depict directional flow, and to determine the relationship between flow dynamics and clinical outcome after decompress surgery. We performed pre- and post-operative MRI in 9 patients with syringomyelia with SPAMM technique. They showed caudal shift of tagging bands in the syrinx cavity in systolic phase while stagnation of CSF flow was seen in diastolic phase. The degree of band shift in syrinx was well correlated with the degree of shrinkage in size of syrinx after operation. Evaluation of syringomyelia by SPAMM technique can predict postsurgical outcome after decompress surgery. PMID- 11854931 TI - Quantities of receptor molecules for colony stimulating factors on leukocytes in measles. AB - We analyzed the comparative amounts of granulocyte-colony stimulating factor (G CSFr) and granulocyte macrophage CSF (GM-CSFr) receptors expressed on neutrophils and monocytes in measles patients to investigate the role of these CSFrs in the development of leukopenia including neutropenia and monocytopenia in measles. EDTA-anticoagulated peripheral blood of 19 measles patients, 10 children with other infections showing leukopenia and 16 children with normal complete blood cell counts (CBC)s were analyzed using flow cytometry and QuantiBRITE. The leukocyte (5260 +/- 2030/uL vs. 9900 + 2680/uL, p=0.000), neutrophil (2580 +/- 960/uL vs. 4250 +/- 2750/uL, p=0.024) and the lymphocyte counts of measles patients (1810 +/- 1430/uL vs. 4530 +/- 3450/uL, p= 0.006) were lower than in the normal controls. The neutrophils of measles patients expressed similar amounts of G- CSFr (1858 +/- 355) as normal children (1764 +/- 477, p= 0.564) and leukopenic patients (1773 +/- 673, p=0.713), but lower levels of GM-CSFr (535 +/- 118) than normal children (957 +/- 344, p=0.000) and leukopenic patients (832 +/- 294, p=0.002). The monocytes of measles patients expressed similar amounts of G-CSFr (916 +/- 336) and GM-CSFr (3718 +/- 906) as normal children (1013 +/- 391 and 4125 (2645, p > 0.05) but less than leukopenic patients (1454 +/- 398 and 5388 +/ 806, p > 0.05). The neutrophil and monocyte counts of measles patients did not correlate with the amount of G-CSFr or GM-CSFr expressed on neutrophils or monocytes (p > 0.05), but in the normal children, the monocyte count correlated with the levels of GM-CSFr on monocytes (r=0.951, p=0.049). In conclusion, neutropenia is one of the more important characteristics of measles patients, which could be due to the decreased GM-CSFr expression on neutrophils. However, the monocytopenia found in measles patients is not due to the decreased expression of CSFr on the monocytes. PMID- 11854932 TI - Endometriosis in an adolescent population: the severance hospital in Korean experience. AB - The records of adolescent patients (10-21), who were admitted to the severance hospital between 1990 and 1999, were reviewed retrospectively to evaluate the age distribution, diagnosis, clinical stage, and treatment for endometriosis in adolescents. Thirty-nine patients who had undergone a laparotomy or laparoscopy and were diagnosed with endometriosis were identified. Endometriosis was classified according to the revised American Fertility Society classification (AFS). The chief symptoms leading to the diagnosis, clinical stage, age distribution, and treatment modality were reviewed. All patients, whose average age of menarche was 14.2, were diagnosed with endometriosis with an average interval of 5.9 years after menarche. The chief symptoms leading to the diagnosis were chronic pelvic pain (27%), acute pelvic pain (21%), a palpable pelvic mass (21%), and dysmenorrhea (18%). A laparoscopy was performed in 20 (51%). The majority of patients (44%) presented with the revised AFS classification stage II. Four patients (10%) presented with stage I, 11 (28%) with stage III and 7 (18%) with stage IV. Management after surgery included GnRH agonists (64.1%), expectant managements (25.7%), OCPs (5.1%) and danazol (5.1%). In adolescents with chronic pelvic pain, endometriosis is not rare. Therefore, early referral to a gynecologist to diagnose the etiology of the pelvic pain and initiate appropriate therapy is recommended. PMID- 11854933 TI - Effect of hysterectomy on conserved ovarian function. AB - The aim of this study was to assess the impact of premenopausal Total Abdominal Hysterectomy (TAH) on the function of the remaining ovaries by reviewing the menopausal age in TAH treated patients. We retrospectively reviewed the medical records of 510 women who had previously undergone TAH, either with or without unilateral salpingo-oophorectomy, due to benign disease at the department of Obstetrics and Gynecology, Yonsei University College of Medicine, between Jan 1989 and Dec 1992. Out of the 510 women, the 94 who were throughly followed up were included in the study, and their menopausal age based on patient symptoms was compared to that of the control group. The mean menopausal age in TAH treated patients was significantly lower than that of the control group (P < 0.001). There was a positive correlation between age at operation and menopausal age. From this study, we could conclude that TAH accelerated ovarian dysfunction, and that the younger the patient was at the time of operation, the earlier the onset of menopause. It is hence apparent that women treated with TAH are at risk of early menopause and should receive adequate hormone replacement therapy. PMID- 11854934 TI - Mutations in the embB locus among Korean clinical isolates of Mycobacterium tuberculosis resistant to ethambutol. AB - Resistance of Mycobacterium tuberculosis to ethambutol (EMB) has been assigned to an operon, embCAB, which has been proposed to be a structural gene for mycobacterial arabinosyl transferases. Recently, genetic events resulting in structural mutations at embB have been proposed as major contributors to the EMB resistance of isolates whose minimum inhibitory concentration (MIC) level is higher than 20 microgram/ml. On the contrary, isolates with a MIC level lower than 20 microgram/ml do not seem to contain any sequence alterations. In this study, in an effort to understand the role of embB mutations at a low-level of EMB resistance, we investigated the sequence polymorphisms of clinical isolates whose MIC levels are lower than 10 microgram/ml. Accordingly, the sequence alterations of a 312-bp region of the embB gene containing the 306th codon, which has been assigned as a hot-spot for EMB-resistance related mutations, were determined for 21 EMB-resistant and 5 EMB-susceptible clinical isolates. In brief, among 21 EMB- resistant isolates examined, 12 (57.1%) contained mutations in embB (10 at the 306th codon and 2 at other sites), and the remaining isolates 9 contained no mutations in any region of embB. The observed mutations included M306V, M306I, and M306L substitutions that have been reported previously. However, 3 were novel types, which included M306T, A313G and Y319C, D328Y [corrected] double substitutions. On the other hand, all of the EMB-susceptible isolates were found to be free of mutations. In conclusion, our findings suggest that sequence polymorphism of embB may play a pivotal role in the EMB- resistance of M. tuberculosis. PMID- 11854935 TI - Assessment of autonomic nervous system with analysis of heart rate variability in children with spastic cerebral palsy. AB - The purpose of this study was to investigate the function of the autonomic nervous system in children with spastic cerebral palsy (CP) through an analysis of heart rate variability (HRV) occurring with orthostatic stress. Twelve children with spastic CP and twelve normal children participated in this study. The echocardiogram (ECG) signals were recorded for 3 minutes in both the supine and 70 degrees C head-up tilt positions, and then the HRV signals underwent power spectrum analysis at each position. Two components were measured; a low- frequency (LF) component (0.05 - 0.15 Hz) primarily reflecting sympathetic activity during orthostatic stress and a high-frequency (HF) component (0.15 - 0.4 Hz) reflecting parasympathetic activity. In the supine position, there was no significant difference between any of the HRV components of the two groups. In the head-up tilt position, absolute and normalized LF were significantly increased and absolute HF was significantly decreased in the normal children (p < 0.05), but not in the children with spastic CP. The results of this study suggest that cardiac autonomic functions, such as vagal withdrawal and sympathetic activation which occur during head-up tilt position, are not sufficient to overcome the orthostatic stress arising in spastic CP children. PMID- 11854936 TI - Histologic changes of pulmonary arteries in congenital heart disease with left-to right shunt (part 1): correlated with preoperative pulmonary hemodynamics. Emphasizing the significance of pulmonary arterial concentration. AB - The hemodynamic findings related to the histologic changes in the small pulmonary arteries in congenital heart disease were analyzed with a left-to-right shunt. A lung biopsy was performed during the repair because of pulmonary arterial (PA) hypertension (mean PA pressure > or = 15 mmHg) in 38 patients. There were 13 patients whose age ranged from 2 years to 25 years old. A preoperative cardiac catheterization was performed to locate the site of the defect and to determine the preoperative hemodynamic findings. Among them, 29 patients had pulmonary vascular resistance (PVR) > 2.5 unit/m2. The lung biopsy specimens were investigated microscopically for the Heath- Edward grade, morphometric analysis of medial wall thickness (MWT) and the rate of the decrease in the pulmonary arteriolar concentration (PAC) obtained by the alvoelo-arterial ratio divided by patients' age. All patients were in the Heath-Edward grade I to III (29 patients in grade I). This grade correlated with the MWT, but did not correlate with a decrease in the PAC. The MWT and the rate of the decrease in the PAC did not accompany each other, but either one had the tendency dominate the pattern in individual patients. The MWT had a close correlation with the mean PA pressure and PVR, and an even closer correlation in patients with a high PVR and those older than 2 years of age. The rate of the decreased in the PAC showed a weak correlation with the shunt volume in patients over 2 years of age or with a large shunt. In the high flow group (PVR < 2.5 unit/m(2), Qp/Qs > 2.0, n=14) the MWT was significantly thinner and the rate of the decrease in the PAC was significantly higher than the high resistance group (PVR > 2.5 unit/m2, Qp/Qs2 < 2.0, n=13). The rate of the decrease in the PAC correlated with the patients' age, but the MWT did not. The lung biopsy results in patients who had both left to-right shunts and pulmonary hypertension showed that the rate of the decrease in the PAC was weakly related to the shunt volume and the MWT was related to the PA pressure and PVR. Either an increased MWT or the rate of the decrease in the PAC tended to dominate. These phenomena were prominent in patients older than 2 in whom a wide range of individual variations were noted in the morphometric pattern. The medial hypertrophy and the rate of the decrease in the PAC may be induced by different stimuli or that medial hypertrophy may play a role in preventing PAC decrease. PMID- 11854937 TI - Histologic changes of pulmonary arteries in congenital heart disease with left-to right shunt (part 2): emphasis on the significance of pulmonary arterial concentration in the correlation with pulmonary hemodynamics after repair Ed-- the above is an alternative title for your consideration. AB - We performed this study to assess the correlation of residual pulmonary hypertension in the immediate postoperative period with that in the late follow up period, to assess the histologic changes of pulmonary arteries (PA) at the time of repair for patients with congenital heart disease consisting of left-to right shunt, and to clarify the role of lung biopsy in determining the operability and reversibility of pulmonary vascular changes. Lung biopsy was performed during repair in 38 patients, with a wide range of age, who had congenital left to right shunt and pulmonary hypertension. All were Heath-Edward grade III or less. Morphometric study included measurement of medial wall thickness (MWT) and decrease rate of pulmonary arterial concentration (PAC). Mean PA pressure in the immediate postoperative period was measured in all 38 patients. Follow-up cardiac catheterization was performed in 15 patients (average 3.8 years after repair). At operation, 5 patients of this late follow-up group were under 2 years of age and the other 10 were 2 or more. During catheterization, pulmonary hemodynamic reaction was observed both under room air inhalation and after inhalation of hypoxic gas FiO2 0.15. Mean PA pressure and pulmonary vascular resistance (PVR) in the immediate postoperative period had a significant correlation with PA pressure and PVR values before the operation, but not with morphometry, Heath-Edward grade, or with pulmonary hemodynamics in late follow-up. During the late follow-up study, 5 of the 15 patients had pulmonary hypertension (defined as mean PA pressure > or = 15 mmHg) under room air inhalation, and PA hypertension was induced in 4 additional patients after hypoxic gas inhalation. There was no incidence of PA pressure or PVR values registering above the preoperative level. The degree of PA hypertension showed a correlation with the rate of PAC decrease and also with patients' age-at operation. Multiple regression analysis showed that both the rate of PAC decrease and the age-at-operation contributed significantly to the degree of PA hypertension. Some of the patients over age 2 had a decreased rate of PAC above the regression line, which none of the patients under age 2 experienced. In patients with Heath-Edward grade III or less, residual pulmonary hypertension in the immediate postoperative period was not correlated with histology, but in late follow-up, it was with PAC and the age-at-operation. Therefore, a decrease of PAC is assumed to be a totally or partially irreversible pulmonary vascular change depending on the patient's age-at-operation, while medial hypertrophy is thought to be a reversible pulmonary vascular change. Lung biopsy could play an important role in determining the reversibility of pulmonary vascular obstruction, particularly in patients older than 2 years. Ed- re highlights above: such hyphenation is optional, but if used then it should be applied consistently throughout the paper. As 3 of the 4 entries in the abstract use it, I have maintained it consistently below. PMID- 11854938 TI - An individualized teaching program for atherosclerotic risk factor reduction in patients with myocardial infarction. AB - This study was conducted to evaluate the effect of a teaching program on patients with myocardial infarction. Forty-five patients were randomly selected 22 were assigned to a teaching group and 23 to a control group. An individualized teaching program was delivered to the teaching group during the hospitalization period. It covered aspects such as: the characteristics of heart disease, the anatomy and physiology of the heart, risk factors of atherosclerosis, medication and diet and exercise therapy. When these subjects were discharged to their homes, they received regular supportive care via telephone or mail for 12 weeks. Atherosclerotic risk factors, including, smoking, exercise, blood lipid profile and BMI were measured before and after the teaching program. Post-testing revealed that the numbers of those who exercised and the number of non-smokers were significantly higher in the teaching group than in the control group. Increased HDL cholesterol (High-Density Lipoprotein cholesterol) was significantly greater in the teaching group than in the control group. The above findings suggest that this individualized teaching program might be helpful at reducing the risk factors of atherosclerosis in myocardial infarction patients. PMID- 11854939 TI - Bone metabolism and bone mineral density in premenopausal women with mild depression. AB - This study was undertaken to investigate the bone metabolism and bone mineral density (BMD) in female patients suffering from depression. Forty-two female patients diagnosed with depression and 42 healthy women, all in the premenopausal age, were enrolled. A clinical evaluation, measurements of the biochemical markers of bone metabolism and BMD measurements were performed. The BMD values were found to be similar in all measured sites. It was concluded that a low BMD was not a prominent feature of premenopausal women with mild depression, even though an increase in bone resorption was found. PMID- 11854940 TI - A case of multiple schwannomas of the trigeminal nerves, acoustic nerves, lower cranial nerves, brachial plexuses and spinal canal: schwannomatosis or neurofibromatosis? AB - In most cases, while schwannoma is sporadically manifested as a single benign neoplasm, the presence of multiple schwannomas in one patient is usually indicative of neurofibromatosis 2. However, several recent reports have suggested that schwannomatosis itself may also be a distinct clinical entity. This study examines an extremely rare case of probable schwannomatosis associated with intracranial, intraspinal and peripheral involvements. A 63-year-old woman presented with a seven-year history of palpable lumps on both sides of the supraclavicular area and hearing impairment in both ears. On physical examination, no skin manifestations were evident. Facial sensory change, deafness in the left ear and decreased gag reflex were revealed by neurological examination. Magnetic resonance imaging revealed multiple lesions of the trigeminal nerves, acoustic nerves, lower cranial nerves, spinal accessory nerve, brachial plexuses, and spinal nerves. Pathological examination of tumors from the bilateral brachial plexuses, the spinal nerve in the T8 spinal position and the neck mass revealed benign schwannomas. Following is this patient case report of multiple schwannomas presenting with no skin manifestations of neurofibromatosis. PMID- 11854941 TI - Acute renal failure associated with a minimal change nephrotic syndrome in a systemic lupus erythematosus patient. AB - In systemic lupus erythematosus (SLE), acute renal failure (ARF) is usually associated with severe lupus nephritis and ARF associated with other glomerular diseases is extremely rare. We recently encountered a patient with ARF that was associated with a minimal change nephrotic syndrome (MCNS) in SLE. A 41-year-old woman presented with a nephrotic syndrome and ARF. She fulfilled four of the American College of Rheumatology criteria for the classification of SLE. However, a renal biopsy revealed that there were no glomerular abnormalities and no deposition of immune complex. The generalized edema disappeared and the high creatinine levels decreased after prednisolone therapy. PMID- 11854942 TI - Emergency endovascular treatment of internal carotid artery injury during a transsphenoidal approach for a pituitary tumor --case report--. AB - Carotid artery injury is a very rare, but life threatening complication that can occur during a transsphenoidal approach. We experienced one case of carotid artery injury during a transsphenoidal pituitary tumor surgery. The patient was immediately treated by a balloon occlusion and complete packing of the cavernous carotid artery using Guglielmi detachable coils (GDCs) and the rest of the tumor was removed after the carotid occlusion. The patient recovered without showing any neurological deficits. PMID- 11854943 TI - Total necrosis of hepatocellular carcinoma due to spontaneous occlusion of feeding artery. AB - Spontaneous total necrosis of hepatocellular carcinoma is extremely rare, with only 15 cases reported to date in the English literature, and the involved mechanism remains unresolved. This paper describes a case of spontaneous necrosis of hepatocellular carcinoma in a 70-year-old man with chronic hepatitis. The patient suffered epigastric pain on admission and computed tomography revealed a 4 cm mass with low density in the left lobe of the liver. Fine needle aspiration biopsy revealed a few scattered, naked and irregular nuclei exhibiting nuclear hyperchromasia in the dirty necrotic background, a finding highly suggestive of malignancy. The lobectomized liver revealed a 3.5 cm, well encapsulated, round, and nearly totally necrotic mass. On microscopic examination, the tumor was found to be composed of thick trabeculae of necrotic tumor cells, supporting the diagnosis of hepatocellular carcinoma. After surgery and throughout 13 months of follow up the patient has recovered well. PMID- 11854944 TI - Successful treatment of cisplatin overdose with plasma exchange. AB - We report a 48-year-old man with laryngeal cancer who received a massive cisplatin toxic overdose without intravenous prehydration through an error in prescription. He received 400 mg/m(2) of cisplatin over a 4-day period. On day 4, he exhibited a broad range of cisplatin toxicities and emergency plasma exchange was started. From day 5 through 19, he underwent 9 cycles of plasma exchange and his plasma cisplatin concentration decreased from 2,470 ng/ml to 216 ng/ml. He completely recovered without any sequelae. No previous reports exist in the English literature of survival without complication after the administration of such a high cisplatin dosage without prehydration. PMID- 11854945 TI - Two primary intracranial tumors of different histology: report of a case with a choroid plexus papilloma and a concurrent vestibular schwannoma in the cerebellopontine angle. AB - A very rare case of multiple primary intracranial tumors is reported. A 41-year old female patient was referred for surgery with a cerebellopontine angle (CPA) tumor. Medical history and MRI study showed typical findings of a right acoustic neuroma with a hydrocephalus. Neurological, dermatological, and ocular examinations revealed no evidence of neurofibromatosis. During surgery, a red colored cauliflower like mass was found in the right CPA. The roof of the fourth ventricle could be seen through the lateral recess after removal of the tumor. Another mass, a 1.5-cm sized schwannoma protruding through the right internal auditory meatus, was removed by the transmeatal approach. Although the tumor masses were in contact and compressed against each other, there was a clear demarcation between them. Histological examination confirmed that the first mass was a typical choroid plexus papilloma with fibrovascular core, and that the second was a schwannoma. The patient recovered without any new neurological deficit. Result of a Medline search indicated that this rare combination of multiple primary tumors has not been reported previously. PMID- 11854946 TI - Shaping a research agenda for the African-American community: issues and opportunities. AB - Building on a previously published NBNA article, this article emphasizes the continuing need for African-American nurses to be actively involved in shaping a research agenda for improving the health and well-being of the African-American community. Thus, this article briefly highlights implications for research targeting African-Americans and summarizes selected information on the National Institute of Nursing Research (NINR). As the National Black Nurses Association and its members continue to develop, implement, and evaluate a research agenda targeting African-Americans, they are encouraged to explore opportunities for research training and career development provided through the National Institute of Nursing Research (NINR). PMID- 11854947 TI - Motivators of health seeking behavior in urban African-American men: an exploration of triggers and barriers. AB - A health crisis exists with African-American men. Morbidity and mortality are higher among African-American men than any other racial/ethnic group. This disparity may be partially related to health seeking behavior in African-American men. Health seeking behavior is influenced by both internal and external motivators. Using the Health Belief Model as a guiding theoretical framework, a qualitative study using focus group methodology was conducted with adult urban African-American men in a large northeastern metropolitan area to explore motivators of health seeking behavior. A total of 38 men were interviewed. Sessions were audiotaped, transcribed verbatim, and analyzed using thematic content analysis techniques. External motivators were identified as resource availability and significant others to include family and associates. Internal motivators were identified as perceived disability and death from an illness and unrelieved symptoms. For most men, seeking care was done after all other measures failed to relieve symptoms. Interventions that will facilitate health seeking behavior in African-American men are needed in order to impact the health disparity. This study identified motivators needed to encourage health seeking behavior in African-American men. The results can be used to develop programs specifically targeting African-American men. Additionally, the study identified key variables for further investigation. PMID- 11854948 TI - Distress and hardiness: a comparison of African-American and white caregivers. AB - The difficulties and burdens of family caregiving, including mental and physical distress, have been well delineated, especially among White populations. Researchers have suggested that buffers for stress associated with caregiving might include hardiness, social support, or cultural group membership, with African-American caregivers perceiving less stress. This paper reports mental and physical distress and family hardiness among 46 African-American and White caregivers of stroke survivors. In contrast to other reports, African-American and White caregivers in this sample were found to be similar on most demographic measures, estimates of caregiver health, estimates of distress, and estimates of hardiness. The single estimate which showed interethnic difference was the confidence sub-scale of the Family Hardiness Index, on which African-Americans scored lower. This sub-scale is believed to estimate confidence in ability to cope with life's problems. PMID- 11854949 TI - Psychological factors influencing cardiovascular risk reduction behavior in low and middle income African-American women. AB - Cardiovascular disease is the number one cause of death for women, especially, African-American women in the United States. In fact, it is reported that each year, 39% of all deaths in women are from coronary heart disease (CHD). Further, 1 out of every 5 women in the United States have some form of cardiovascular disease (CVD). In 1998, 500,037 women died from CVD. In 1997, the morality for CHD for females was 236,468; representing 491 of all deaths for this disease. Among women, African-American women have a death rate for CHD almost four times higher than their White counterparts. Several factors influence the risks for African-American women. African-American women are more likely to be obese and to participate in lower rates of cardiovascular risk reduction behavior (diet control, exercise, weight management) than Caucasian women. Few studies explore psychological factors influencing African-American women's participation rates in cardiovascular risk reduction behavior. The purpose of this study was to explore the role of psychological factors (self-concept, self-efficacy to diet and exercise, and stress) on cardiovascular risk reduction behavior in low and middle income African-American women. A questionnaire measuring self-concept, self efficacy to diet and exercise, and stress was self-administered by middle-income women and investigator administered to low-income women. Findings revealed that self-efficacy to diet and exercise were positively correlated with cardiovascular risk reduction behavior in both groups. Additionally, middle-income women with higher levels of self-concept and lower body mass index were more likely to participate in the risk reduction behavior. Findings have implications for tailoring cardiovascular risk reduction interventions to African-American women of varying socioeconomic status (SES). PMID- 11854950 TI - Reduction of high-risk behaviors among incarcerated youths: a challenge in prevention. AB - The adolescent population in America is at increasing risk for acquiring the human immunodeficiency virus (HIV) that causes AIDS (CDC, 1998; 1996; 1994; 1993). Although the total number of reported AIDS cases among adolescents 12 to 19 years of age is, thus far, fairly small, the identified caseload in this age group has increased by over 40% during one year alone. In fact, one-fifth of all reported AIDS cases occur among persons 20 to 29 years of age. Many of these young adults contracted HIV infections as teenagers (CDC, 1996). But the trend is changing. In fact, among younger youths, especially those of African heritage, the infection rates are rapidly increasing (CDC, 1999; CDC, 1998). These increases will require focused attention by all health professionals, and public policy officials (Lai, Tsai, Hardy, 1997; Porter, Oakley, Guthrie & Killion, 1999). The purpose of this current study was to describe risk-related behaviors of incarcerated youths that are significant factors when planning community-based programs for HIV/AIDS prevention. Several research questions were generated in this regard and tested. Several significant findings emerged from the data that indicate that African-American youths incarcerated are at greater risk for development of HIV/AIDS than their other ethnic counterparts. PMID- 11854951 TI - Saving our children: strategies to empower African-American adolescents to reduce their risk for HIV infection. AB - Adolescence is normally a healthy period of life. For some young people it is a period of experimentation with risky behavior. For others, it marks the development of habitual risk behaviors that persist into adulthood. Of special concern is adolescent involvement with sexual behaviors that increase the risk of infection with HIV and other sexually transmitted diseases (STDs). Nurses who work with adolescents are seeing an increase in STDs, including HIV infection occurring disproportionately among African-American adolescents. Although the use of condoms can reduce the risk of these sexually transmitted diseases, most sexually active adolescents do not consistently use condoms. This paper will discuss the scope of the problem of STDs, especially HIV infection among African American adolescents. It will describe the Theory of Planned Behavior as a framework for designing interventions to reduce the sexual transmission of HIV and other STDs. Finally, it will provide strategies for nurses to intervene by empowering African-American adolescents to reduce their risk for sexually transmitted HIV infection. PMID- 11854952 TI - The perceptions of menopause of African-American and white women and affect on willingness to participate in a HRT clinical trial. AB - Menopause is a normal developmental process for women; however, few studies of healthy menopausal women and African-American women have been reported. While hormone replacement therapy (HRT) has been touted as a treatment for menopausal women, clinical trials of HRT still need to be conducted to determine its potential positive and negative effects. This is a focus group study of 21 women of menopausal age (11 African-Americans and 10 White women) to determine their experiences with menopause and their willingness to participate in a HRT clinical trial. Results indicated generally positive attitudes toward menopause and a variety of symptoms of varying degrees of severity in African-American and White women. Limited knowledge about menopause and HRT were apparent; however, White women were more likely to be receiving HRT and were more informed about HRT than African-American women. White women also were more positive about participating in a HRT clinical trial than African-American women who feared cancer as a potential side effect. PMID- 11854954 TI - Developing culturally competent care techniques to assist African-Americans to survive in the 21st century. PMID- 11854953 TI - Promotion of sexual health in the American cultural context: implications for health promotion in school age African-American girls. AB - African-American girls are at greater risk of engaging in sexual behavior at earlier ages, and putting themselves at risk for pregnancy and sexually transmitted disease. It may also lead to more limited successful life trajectory. The purpose of this article is to present an overview of some of the issues facing African-American girls today as they leave preadolescence and enter early adolescence. Research and health promotion strategies are identified that need to be examined and addressed when providing care to this population. PMID- 11854955 TI - A quiet migration: cultural influences impacting children adopted from the former Soviet Union. AB - When children immigrate to another culture, a variety of salient variables influence their adaptation. Among internationally adopted children, these variables include early history, changes in socioeconomic status, awareness of multiple issues impacting the child post-adoption, availability of social networks, and anti-immigration sentiments in the host culture. The change in socioeconomic status of children adopted from the former Soviet Union is a positive influence with the concomitant improvement in the child's nutrition and health care. Many internationally adopted children have spent time in institutional environments that constitute a culture in and of itself. The orphanages are microenvironments which may be closed to the influences of the larger culture. The environmental components of the orphanage transmit a collection of traits and values which, taken as a whole, constitute the orphanage culture. Adoptive parents must facilitate the children's adjustment to a new cultural milieu with different language, food, and customs. Further, children must adapt to the culture of the family, a culture that may have been previously unknown. Nurses can promote the adaptation of these families by increasing awareness of the multitude of issues that impact internationally adoptive families. PMID- 11854956 TI - Descriptive study of hypercholesteremia in a Vietnamese population of the Gulf coast. AB - The purpose of the study was to determine the prevalence of high cholesterol and related heart disease risk factors and knowledge, in a selected Vietnamese population. A convenience sample of 114 adult Vietnamese volunteers, who are living in a selected Southern Mississippi Vietnamese rural community, were tested. Only 29% scored at the low risk level for knowledge of cardiovascular health risks. High blood pressure was found in 44% of the sample, with 20% referred for medical follow-up. Of the 58% who had high total cholesterols, 65% had high HDLs, and 35% had high triglycerides. Approximately half of the sample stated that they did not see a doctor annually or on a regular basis. PMID- 11854957 TI - Culturally competent care for members of sexual minorities. AB - Culture has historically been interpreted as the beliefs, mores, and lifeways of groups of people primarily related to race and ethnicity. However, individuals who self identify as being lesbian, gay, bisexual and/or transgendered experience ethnocentrism when seeking care from medical and health professionals. Using the principles and concepts of Lenninger's theory of Culture Care Diversity and Universality, members of sexual minorities can assist their health care providers to provide culturally sensitive and ethical care. PMID- 11854958 TI - Developing community health nursing skills collaboration in a culturally diverse population. AB - The graduate curriculum in Community Health Nursing at the University of South Alabama was revised to prepare nurses to function as a community health specialist. The revised curriculum model includes two semesters that focus on a specific population or high risk group of patients or clients. Emphasis is placed on the skills that community health nurses must have in order to assess communities, identify community needs, plan and implement interventions at the population aggregate or community level. During the first semester, or the practicum course, a community needs assessment is performed. In the internship course, the planned intervention is implemented and evaluated. The purpose of the paper is to describe the process of identifying and accessing a Cambodian population aggregate in a rural setting. The collaboration among faculty, student, preceptor, official agency, as well as lay leaders in the Cambodian community is described. PMID- 11854960 TI - Radiofrequency ablation of liver tumors: the complementary roles of the clinic and research nurse. AB - This article describes a clinical research protocol designed to determine the value of Radiofrequency Ablation (RFA) for the treatment of histologically proven primary or metastatic liver cancer. RFA is a localized thermal technique that destroys tumor tissue. The research protocol is described, including enrollment criteria, performance of RFA, and the follow-up necessary to ascertain protocol success or failure. The complementary and collaborative roles of the research nurse and clinic nurse in carrying out the RFA protocol are described. PMID- 11854959 TI - Intestinal transplantation in pediatric patients: a nursing challenge. Part 2: Intestinal transplantation and the immediate postoperative period. AB - Intestinal transplantation offers new hope to children with intestinal failure and life-threatening complications of parenteral nutrition. As more transplant centers are adding intestinal transplantation to their existing programs, new challenges are presented to nursing professionals. This three-part series will provide information for nursing professionals regarding evaluation for intestinal transplantation, donor preparation, the surgical procedure, immediate postoperative care of the transplant recipient, and long-term care of the intestinal transplant recipient. This article focuses on donor selection and preparation, the surgical procedure for intestinal transplantation, the immediate postoperative course, immunosuppression, rejection, infections, fluid and electrolyte support, and nutrition. The information presented represents the approach taken at University of Nebraska Medical Center/Nebraska Health Systems. PMID- 11854961 TI - Endoscopic mucosal resection. AB - Endoscopic mucosal resection (EMR) is a major advance in endoscopy for treatment of patients with superficial esophageal, gastric, or colonic lesions, providing a nonsurgical treatment option for management of these lesions. With the assistance of endoscopic ultrasonography, it is now possible to obtain an accurate histologic diagnosis, confirm the depth of the lesion, and in many cases resect submucosal tumors. The main goal of EMR using the advances in endosonography is to detect early gastrointestinal cancers and successfully resect them, offering an outpatient, nonsurgical treatment option. Although popular in the Orient, where there is a high incidence of superficial neoplasia, limited data are available on the use of EMR in the United States. Gastrointestinal (GI) endoscopy nurses and assistants play important roles in successful EMR. This article informs GI staff on the indications for EMR, the procedure and accessories needed, the different resection methods, possible complications, and nursing care. PMID- 11854962 TI - Benchmarking: your performance measurement and improvement tool. AB - Many respected professional healthcare organizations and societies today are seeking to establish data-driven performance measurement strategies such as benchmarking. Clinicians are, however, resistant to "benchmarking" that is based on financial data alone, concerned that it may be adverse to the patients' best interests. Benchmarking of clinical procedures that uses physician's codes such as Current Procedural Terminology (CPTs) has greater credibility with practitioners. Better Performers, organizations that can perform procedures successfully at lower cost and in less time, become the "benchmark" against which other organizations can measure themselves. The Better Performers' strategies can be adopted by other facilities to save time or money while maintaining quality patient care. PMID- 11854964 TI - Implications of complementary health care practices for endoscopy and GI procedures. PMID- 11854963 TI - Latex allergy and the healthcare worker. AB - The purpose of this article is to describe the clinical manifestations and treatment of latex allergy. A discussion of the identification of latex sensitized staff and available diagnostic tests is presented. Emphasis is placed on the healthcare worker's role in preventing future latex allergy cases from developing. PMID- 11854965 TI - Nurses with political savvy: multi-state licensure. PMID- 11854966 TI - Managerial ethics: arriving at decisions we can live with. PMID- 11854967 TI - Expanding our territory. PMID- 11854968 TI - Do you want to be an author? Ten steps for successful writing. AB - At some point in our lives, we often envision ourselves in other roles. Being an author is usually a fleeting thought. Nurses have the opportunity to share their professional expertise, the latest research information, or stories of human kindness and compassion. You may often intend to write, but lose the motivation when you think that you are not really capable. Although you may never attain the level of Shakespeare, James Joyce, or Mark Twain, you can still have the satisfaction of seeing your article in print. All it takes is a little organization. This article provides an overview of 10 steps to help you get started with writing. These simple key points may not get you the Pulitzer prize, but they can get you started on a new hobby and facilitate important contributions to our nursing discipline through the written word. PMID- 11854969 TI - Irritable bowel syndrome: causes and treatment. AB - Irritable bowel syndrome (IBS) is a common health care problem worldwide. In the United States and Northern European countries, more women than men seek health care services for IBS. Nurses are often called on to help women with IBS manage their symptoms. This article reviews the literature related to gender differences in diagnosis, symptoms (gastrointestinal, somatic, and disturbed sleep), and physiologic and psychological factors as well as current pharmacologic therapies used in the management of IBS. PMID- 11854970 TI - Flexible endoscopes: structure and function--the air and water system. AB - Flexible endoscopes are complex medical instruments that are easily damaged. To maintain the flexible endoscope in optimum working condition, the user must have a thorough understanding of the structure and function of the instrument. This series of articles will present an in-depth look at the care and handling of these expensive devices. The flexible endoscope is constructed of several systems that operate simultaneously to produce a highly technical, yet effective diagnostic and therapeutic medical device. These systems include the air and water system, the suction and operating channel system, the mechanical system, the endoscopic retrograde cholangiopancreatography (ERCP) elevator system, the optical system, and the electrical system. This first article in a series will focus on the air and water system of the endoscope. A review of the internal and external structure of the flexible endoscope and the functions of the air and water system, including infection control issues, potential problems and evaluation, and prevention of minor problems to avoid expensive repairs, will be addressed. PMID- 11854971 TI - Understanding the research methodology: should we trust the researchers' conclusions? AB - Understanding the methodology section of research articles is difficult for those unfamiliar with research design, but is essential for judging the credibility of the researchers' findings. The methodology section describes measures taken to avoid reaching the wrong conclusions. When comparing two or more treatments, researchers might find a difference in treatments when none exists (type I error) or not find a difference when there actually is one (type II error). Type I errors are minimized by random assignment to treatment and control groups, double blinding, ensuring that the treatment under study is the only difference between groups, and setting the level of significance in advance. Type II errors are minimized by using an adequately sized, homogenous sample, increasing the treatment strength, measuring precisely, and using appropriate statistics. The purpose of this article is to explain these common experimental strategies and statistical terms. PMID- 11854972 TI - Monitoring depression in patients undergoing alpha-interferon and ribavirin therapy for hepatitis C. AB - Individuals with hepatitis C virus (HCV) constitute a growing segment of the US population, with most new infections attributable to intravenous drug use. Commonly, there is a 10- to 30-year delay from time of infection to diagnosis. Current treatment is with interferon, alone or in combination with ribavirin. A concerning side effect of both monotherapy and combination therapy is depression, which can become severe and lead to suicide. In patients with liver disease and those who have used intravenous drugs, depression is highest among those who are also positive for HCV. Use of a standardized short form depression self-rating tool would provide the advantages of increased accuracy in patient assessment, improved documentation, and cost-effective monitoring of depression in patients with HCV receiving interferon/ribavirin therapy. This article discusses the importance of screening and monitoring patients for depression as they undergo treatment for HCV infection with interferon alone or in combination therapy with ribavirin. PMID- 11854973 TI - Sedation and analgesia. PMID- 11854974 TI - Performance of flexible sigmoidoscopy by registered nurses for the purpose of colorectal screening. PMID- 11854976 TI - The power of "the vote". PMID- 11854975 TI - Endoscope protection. PMID- 11854977 TI - Helping new nurses succeed. PMID- 11854978 TI - Taking "Healthy People 2010" seriously. PMID- 11854979 TI - Admission: a crucial point in relationship building between parents and staff in child psychiatric units. AB - The admission of a child to a psychiatric unit is difficult for parents. While many articles about admissions can be discovered in the literature, few address the process of admission. A grounded theory study was undertaken to describe and explain the relationships that develop between parents and nurses in child psychiatric units. The crucial nature of the admission process was discovered and is the focus of this article. Theoretical sampling was used. Data were collected by interviewing parents and the nursing staff working with the parents. In addition, participant observation was used. Data were analyzed using the constant comparative method. Parents often came to the unit expecting to be blamed for their child's problems. When nurses responded with reassurance and caring, the parents' engagement was enhanced. During admission, expectations of parents and nurses influenced the relationship. In addition, the study discovered ways that nursing routines influence the relationship during the admission period. PMID- 11854980 TI - Women's midlife confusion: "why am I feeling this way?". AB - Midlife is a multifaceted stage of woman's development, characterized by important transitions. In an ethnographic study of women's midlife experience of their changing bodies, 11 participants voiced their uncertainty and confusion around bodily changes, responses exacerbated by the lack of consistent health related information in this area. This confusion emerged as one of the major thematic elements of the study. Midlife women's experience of confusion may reflect a much broader problem, the locus of which is not so much in the women themselves, but rather in negative societal attitudes about aging women. This article describes various aspects of confusion that emerged from the data, and offers implications of the research and recommendations for practitioners. PMID- 11854981 TI - Perceptions of problems in people hospitalized for bipolar disorder: implications for patient education. AB - This study explored the perceptions of people hospitalized for bipolar disorder in regard to their difficulties in functioning and the most important problem with which they would like the hospital's help. One-hundred-twenty-two patients diagnosed with bipolar disorder completed the Behavior and Symptom Identification Scale (BASIS-32) at the beginning of their hospitalization. The relationships between subjective distress (measured by the BASIS-32 scores) and background characteristics were examined. In addition, participants' perceptions of their most important problems were coded as (1) psychiatric problems, (2) social or physical problems, or (3) no problems, and examined with respect to background characteristics. Race, admission status, and a secondary diagnosis of a substance use disorder were significantly related to overall subjective distress; a substance use disorder diagnosis was significantly related to all five BASIS subscale scores. No background variable was significantly related to the problems with which participants reported wanting the hospital's help, although admission status and race were of borderline significance. PMID- 11854982 TI - Loneliness and depression in caregivers of persons with Alzheimer's disease or related disorders. AB - Secondary analysis of data from a sample of 242 husbands, wives, and daughters providing care for Alzheimer's disease family members was conducted to examine the relationships among loneliness and depression and the following variables: quality of the past relationship, relational deprivation, quality of the current relationship, and distance felt due to caregiving. Loneliness was significantly related to depression (r = .66, p < .001), relational deprivation (r = .36, p < .001), and quality of the current relationship (r = .34, p < .001), indicating that the more loneliness reported by the caregivers, the more the caregiver experienced depression, relational deprivation, and a poorer quality of the current relationship. Significant gender differences were found with the caregiving wives and daughters reporting higher mean scores than caregiving husbands on relational deprivation, loneliness, and depression. Loneliness was the only variable significant for predicting depression in caregiving husbands, wives, and daughters. In order for loneliness and depression to be addressed in the Alzheimer's disease caregiver, they must first be recognized by nurses. PMID- 11854983 TI - A progressive relaxation intervention at the worksite for African-American women. AB - This study examined the efficacy of a worksite-based intervention in reducing blood pressure and personal strain in African-American women. The experimental group (n = 22) was taught a 7-muscle group progressive relaxation intervention. The women, for home practice, used audiocassette tapes and diaries. The control group (n = 21) was asked to set aside 30 minutes for relaxation. All women who participated in the study were visited at the worksite weekly for eight weeks. At the conclusion of the study, the experimental group had a significantly lower mean physical strain score (23.23 [SD = 6.85] v. 27.41 [SD = 8.49]) and interpersonal strain (25.81 [SD = 6.25] v. 23.57 [SD = 6.06]); both groups had significant reductions in psychological strain scores. Although there were no significant differences in the blood pressure of the two groups at the end of the 8-week intervention (121.95 [SD = 13.28] v. 129.33 [SD = 14.30]), the mean systolic blood pressure of the treatment group did decrease from 130 mmHg to 121 mmHg. Findings from this study support the use of the work-place as a means of improving the cardiovascular health of working African-American women. PMID- 11854984 TI - Health care perceptions of mid-western urban-dwelling African-American women. AB - As a part of a comprehensive women's health needs assessment performed in Missouri, two focus groups were conducted with urban-dwelling African-American women. Ten African-American women 18 to 39 years of age comprised one group and 10 African-American women 40 to 74 years of age comprised the other group. Participants discussed major health concerns and issues related to service access. Although similar responses occurred between groups, major concerns such as reproductive issues in the younger women and chronic diseases in the older women, were clearly age-related. Participants commented on impediments to health care and adequacy of services. The Missouri Department of Health will use these findings in planning group-specific health interventions and services. PMID- 11854985 TI - Hope and coping in HIV-infected African-American women of reproductive age. AB - African-American women of reproductive age, particularly those living in the southern United States, represent a disproportionate number of women with HIV/AIDS. Often women who become infected with HIV in the South must cope not only with the disease, but with economic stresses and racial stigma, as well. Faced with these changes, these women experience increased distress and loss of hope. The purpose of this study was to examine levels of hope in a group of HIV infected African-American women and to identify coping strategies that were associated with women maintaining a sense of hope for the future. A descriptive design was used to test the relationships between hope and coping strategies used by HIV-infected African-American women. Reported levels of hope were significantly lower for study participants than for women with breast cancer or for hospitalized adults with cancer who were receiving chemotherapy. Statistically significant positive relationships were observed between hope and the total coping score (r = .37, p = .009) hope and managing the illness (r = .47, p = .001) and between hope and spiritual activities (r = .40, p = .004). A statistically significant negative relationship was observed between hope and avoidance coping (r = -.35, p = .009). PMID- 11854986 TI - Sleep apnea symptoms, nocturia, and diabetes in African-American community dwelling older adults. AB - The primary aim of this exploratory study was to examine the association between obstructive sleep apnea (OSA) symptoms, nocturia and diabetes in African-American community-dwelling older adults. A pencil and paper survey was used to collect symptoms of OSA, nocturia, excessive daytime sleepiness (EDS), lower urinary tract symptoms (LUT), and self-rated health. The convenience sample of community dwelling African-Americans (n = 87) included a majority of women (61%) and obese persons (60% with BMI > 30). The mean age was 64 years (range 50-91), and 40% were self reported Type 2 diabetics. Diabetics had significantly more OSA symptoms, more nocturia, and decreased self-rated health. Results of a stepwise logistic regression showed that persons with diabetes and those who reported EDS experienced significant three to four-fold risks for OSA. Similarly, diabetics, women, persons with LUT symptoms and those reporting EDS, had significant three to six-fold risks for nocturia > or = 2/night. PMID- 11854987 TI - Health promotion for urban middle school students: a survey of learning needs. AB - Community based education programs and community partnerships are crucial for attaining the objectives of Healthy People 2010. The purpose of this descriptive study was to determine the health promotion needs of urban middle school students from the perspective of the participants in a health promotion partnership project. A convenience sample of 161 urban middle school students participated in the study. A Likert scale of health promotion topics was used for data collection. Findings from the study indicated that urban middle school students have a major interest in finding out more about hair care, safety issues, and prevention of infection. Students expressed a need for more information about issues related to why people use drugs, feelings about self, feelings of sadness and worry, and feelings about death. Important variations were found in the priority of interest in topics among the three grades. The researchers concluded that the psychosocial health promotion needs of urban middle school students should be explored further. PMID- 11854988 TI - Multiculturalism as a concept in nursing. AB - This article focuses on the theoretical processes of identifying and evaluating concepts in the public domain and the subsequent practical application of such concepts to the provision of nursing care. More specifically, this article identifies the concept of "multiculturalism" as a valuable one for nursing, and then evaluates it via the methodology developed by nursing theorists Wilson (1969) and Meleis (1997). PMID- 11854989 TI - Fat distribution and cardiovascular disease risk in African-American women. AB - We have previously shown that in Caucasian men and women fat in the trunk, and especially visceral fat, was related to cardiovascular disease (CVD) risk even after adjusting for fat in other depots. We also found that leg fat was negatively related to CVD. In order to determine if these relationships also exist in African-American women, blood lipids, insulin, and blood pressure in 26 pre-menopausal African-American women were evaluated. In addition, magnetic resonance imaging measured fat distribution was evaluated from the lateral malleolus to top of the shoulder. Percent fat was related to VLDL cholesterol, VLDL3 cholesterol, triglycerides, insulin, and diastolic blood pressure. Inclusion of visceral and trunk fat together in multiple regression did little to improve the relationship with CVD risk. Leg fat, however, tended to be negatively related to CVD risk after adjusting for trunk fat and visceral fat. Partial correlations indicated that leg fat was negatively related to VLDL cholesterol (r = -0.36), triglycerides (r = -0.36), insulin (r = -0.46), and diastolic blood pressure (r = -0.43). These results indicate that in African-American women, visceral fat may be less atherogenic than in Caucasian men and women since it was poorly related to CVD risk after adjusting for fat in the trunk. In addition, consistent with results from Caucasians, leg fat is inversely related to CVD risk after adjusting for fat in other parts of the body. Caution must be made in the interpretation of correlational data. Further research is warranted to explore these intriguing relationships. PMID- 11854990 TI - And still we rise: meeting the challenges of ending health disparities among African-Americans. PMID- 11854991 TI - Rationing hospital services in Hong Kong: priority setting by clinicians using the Delphi method. AB - This article presents the results of a demonstration project of priority setting by clinicians in Hong Kong's public hospitals. Thirty-five chiefs-of-service of public hospitals in Hong Kong participated in a three-round Delphi exercise on identifying interventions that should be charged in public hospitals on the basis of questionable effectiveness and other reasons. A total of 246 interventions were identified in the first round, 126 of which received a positive score (indicating a high degree of consensus amongst the participants) at the end of the final round. The interventions that received higher scores tend to be preventive services, treatment of an elective nature, and procedures for sex related conditions. As the number of interventions with positive scores in the final round are relatively small, the amount of money to be recovered from charging these interventions is not likely to be substantial. Such results suggest that rationing by itself cannot be a solution to the problems of healthcare financing in Hong Kong. PMID- 11854992 TI - Colonizing the new world of NHS management: the shifting power of professionals. AB - This paper explores the changing patterns of professional power and the struggle for control between doctors and managers in the UK NHS, by examining the role of clinical directors. Located at the nexus of managerial and professional power, clinical directors represent and embody the challenges to medicine through increased managerialism and the profession's response to it. An analysis of the role of clinical directors reveals the changes in power and jurisdiction that have been created through clinical management. A medical model of professional power illustrates how structural and ideological changes threaten medical dominance. However, clinical directors respond to the changes by creating new forms of expertise through managerial assimilation, to extend their jurisdiction and domain within the organization and in the market. This re professionalization, rather than de-professionalization, by doctors raises questions about the shifting power balance between doctors and managers in the NHS and between doctors within the medical profession. PMID- 11854993 TI - Emerging factors shaping the future of the Veterans' Health Administration: a strategic analysis. AB - In response to societal and industry-wide forces, the Veterans' Health Administration (VHA) has undertaken a re-engineering process, changing the operational and management structure from individual, independent, and often competing large hospital centres into 22 integrated service networks or VISNs to provide structural incentives for efficiency, quality and improved access as well as transitioning the system to one that is grounded in ambulatory and primary care (Ashton et al., 1998). This paper presents a framework for evaluating the successes and/or failures of the recent re-organization efforts of the VHA in bringing together this multitude of medical care 'parts' or modules into an integrated, cost-effective healthcare delivery system. In total, this paper attempts to delineate an analytical framework by which the threats and opportunities as well as the strengths and weaknesses of the VHA are identified. More specifically, this paper addresses the external pressures driving reform in the VHA system and how the Veterans' Administration can respond to these pressures. Implications for the future of the VHA if its reform efforts are not successful are examined. PMID- 11854994 TI - Impact on health authorities of the introduction of primary care groups and trusts. AB - The National Tracker Survey of Primary Care Groups (PCGs) and Trusts (PCTs) in the UK has provided evidence about how PCGs and Health Authorities managed the transition to PCTs. This was a major challenge for both organizations. Devolution of responsibilities was dependent on the capacity and readiness of PCGs to take on their new roles. The development of good working relationships between the principal parties was critical to this process. Health Authorities started to let go and all put in place basic mechanisms for holding PCGs to account in their first year. However, a number of PCGs regarded their Health Authority as authoritarian. Most Health Authorities were found to have started to provide information to support PCGs in their core functions, but resources were frequently problematic. It will be important for Health Authorities and PCG/Ts to negotiate a shared view of their respective roles and responsibilities. The future role of Health Authorities in providing strategic leadership for their local health economies was not sufficiently well defined or understood. PMID- 11854995 TI - Public versus private hospital maternity length of stay: a gamma mixture modelling approach. AB - Application of a gamma mixture model to obstetrical diagnosis-related groups (DRGs) revealed heterogeneity of maternity length of stay (LOS). The proportion of long-stay subgroups identified, which can account for 30% of admissions, varied between DRGs. The burden of long-stay patients borne was estimated to be much higher in private hospitals than public hospitals for normal delivery, but vice versa for Caesarean section. Such differences highlights the impact of DRG based casemix funding on inpatient LOS and have significant implications for health insurance companies to integrate casemix funding across the public and private sectors. The analysis also benefits hospital administrators and managers to budget expenditures accordingly. PMID- 11854996 TI - Policies, priorities, opportunities and barriers in mental health services: five years of the London managers' survey. AB - Since 1996, an annual survey of senior managers of mental health services in London has been undertaken in order to identify trends in their views of their task. This is the only longitudinal study in the current literature of the views of a group of health and social care managers in England. This paper first outlines the methods and the findings of the survey in 2000, drawing comparisons with data collected in 1998 and 1999 (and occasionally earlier). The discussion section draws out the central themes that emerge from the surveys: firstly, community mental health teams hold a central role in the mental health system; secondly, the development of trust between agencies is a complex challenge; thirdly, it appears that social care managers may be more committed to implementation of national policy than their health colleagues; fourthly, personal and organizational instability is a fact of life; and finally, the relationship between the promulgation of national policy and its adoption into local practice is not straightforward. The conclusion reflects on the potential implications of these findings for the future development of mental health services in England. PMID- 11854997 TI - Contemporary nurse--who are you? PMID- 11854998 TI - Virtuous principles as an ethic for nursing. AB - A theoretical analysis of virtuous principles--beneficence, non-maleficence, autonomy and justice provides a response to the question: 'How ought ethical nursing practice be characterised?' These virtuous principles promote human flourishing and sustain and strengthen nursing practice. Beneficence, described as acting to benefit others, incorporates a balance of benefit over harm and requires nurses to take action. Non-maleficence calls nurses to refrain from inflicting harm and defines prohibitions over actions. Autonomy, defined as self governance and self-determination, may have limitations so that absolute or perfect autonomy might be unrealistic. Finally, Justice, a problematic concept, is aligned with fairness and what a patient deserves. PMID- 11854999 TI - Is competency based training/education useful for workplace training. AB - This paper discusses and explores whether competency based training is useful for workplace training and education. Competency based training and education has re emerged over the past decade as the panacea to meet the education and training needs to develop a more flexible and skilled Australian workforce. Nurse education has been affected by these re-emerging trends. The historical developments associated with competency based training is presented. Specifically the relevance of competency based training to nursing and nurse education is examined. PMID- 11855000 TI - Factors impacting on shared care working relationships between practice nurses and general practitioners: a literature review. AB - Practice nurses have worked in general practice for many years yet little is known or published in Australia about their work. Recent health care reforms in Australia, particularly the establishment of the Divisions of General Practice and various best-practice innovations linked to the National GP Strategy have seen an increase in the role of practice nurses. Despite this GPs in Australia are cautious about the role of practice nurses. Much of this caution arises from reform in general practice in Britain particularly the establishment of fund holding for general practice. This article reviews the literature on practice nurses in Britain and Australia within the framework of the health reforms in general practice in both countries. PMID- 11855001 TI - The educational needs of part time clinical facilitators. AB - Clinical facilitators play an important role in the clinical practicum component of an undergraduate nursing programme. Part time teachers are often employed in this role, however very little has been written about the needs of these facilitators. An exploratory study design incorporating focus group interviews and questionnaires was used to identify the needs of clinical facilitators at a large Australian university. This study showed that they are committed nurses who often feel inadequate in a role they feel lacks support and identity. The findings of the study have implications for the preparation of clinical facilitators. PMID- 11855002 TI - The pain of redundancy: an exploration of the impact of redundancy for nurse managers in Victoria in the 1990s. AB - This article reports on research conducted in 1995 in Victoria, Australia with 12 nurse managers who were displaced through redundancy. This was a pilot study that employed a phenomenological approach to explore the personal impact of redundancy. Participants experienced three differing approaches to the redundancy processes undertaken. Data was managed by a thematic analysis. Powerful themes of being isolated in their agony and fear, of consent under duress, and trauma, emerged from the data. This paper tells the story graphically and emotionally, it is the people's story, an eloquent example of what is missed in current literature. PMID- 11855003 TI - Nurses management of post-operative pain. AB - Nurses have the responsibility of adequately managing patients' post-operative pain. This literature review assesses whether nurses' management of post operative pain is adequate or not, according to the literature findings. The findings reveal that nurses' management of patients' post-operative pain is not adequate and implies the concurrent need for improved nurse education and practice. The findings also indicate a need for ongoing research of this phenomenon. PMID- 11855004 TI - Comparing Norton, Braden and Waterlow risk assessment scales for pressure ulcers in spinal cord injuries. AB - When pressure ulcers occur health care services are faced with considerable challenges and costs. Additionally these ulcers cause significant physical and psycho-social impairment for patients and their families. The complexity of pressure ulcer management is widely acknowledged in the literature. However, the applicability of the major risk assessment scales in the context of spinal cord injury has not been explored. Retrospective case history audit of individuals with pressure ulcers provided data to compare the three commonly used Norton, Braden and Waterlow scales. Waterlow's scale was the most sensitive of the three scales compared. PMID- 11855005 TI - Client perceptions of the polysomnography experience and compliance with therapy. AB - A grounded theory approach was used to generate theory about clients' perceptions of polysomnography, and the relationships between these perceptions and compliance with therapy. Interviews were conducted with two groups of clients: those who complied with recommended therapy (Continuous Positive Airways Pressure) and those who did not. Compliance with suggested therapy appeared to be related to the degree of benefit derived by the client. In addition, there were problems with the process of diagnosis and follow-up that impacted on client satisfaction and may have influenced their compliance. These findings have highlighted issues which have been used to generate theories that will be tested in future research. PMID- 11855007 TI - A guide to the use of focus groups in health care research: Part 2. AB - In this paper, the second in a two part series on focus groups, a comprehensive overview of the literature concerning the interviewing process, methodology and analysis is provided. We discuss the characteristics of the moderator and the way in which a focus group is conducted (the who, where, and how of the focus group process), the participants, the process, and the analysis. Nursing staff and allied health professionals, especially those contemplating using focus groups for the first time, will be able to gain an understanding of the conduct of focus groups from this one source. The comprehensive reference list will provide a wealth of primary sources for further reading. PMID- 11855006 TI - A guide to the use of focus groups in health care research: Part 1. AB - Focus group interviewing is becoming a popular research method among nurses as well as allied health professionals. Prior to performing a focus group interview a novice must first search the literature to gain an understanding of the function, uses, preparation, conduct and analysis of this form of methodology. This paper has been compiled in an attempt to synthesise, condense and simplify the information from an extremely wide and varied focus group literature and provide a comprehensive review to assist nurses who are considering utilising this methodology in their research studies. The purpose of this paper was stimulated by one of the author's (MP) own experience learning about focus group methodology before commencing her first focus group. PMID- 11855008 TI - Culture and diversity in the nursing classroom: an impact on communication and learning. AB - This article discusses culture and diversity in the nursing classroom and its impact on communication and learning. Today's nursing classrooms are heavily populated with students from many ethnic, psychological and sociocultural backgrounds. It is necessary that nurse educators recognize that many communication patterns value diversity, and is a major support in many learning styles of their students. Without a thorough understanding of diversity and communication among cultures, major challenges exist for both professors and students. The nursing curriculum and classroom materials should portray diversity so that all subject matter reflects a range of cultural perspectives. Higher education curricula are probably the most diverse in the world, and with a global perspective, and the United States being a "melting pot," these curricula will become even more diverse in the future. Nursing education has the responsibility to teach students how to communicate on a global scale. These global models of communication will assist students to successfully enter professional nursing practice with strong communication skills and a level of cultural competence that professional nurses need to know. PMID- 11855009 TI - Substance abuse in African American women. AB - Substance abuse is a serious problem from which, regardless of sex or race, no one is immune. Each racial and gender group has specific etiological factors relating to the use of illicit drugs. Data regarding substance abuse in African American women has only recently begun to emerge in the literature. Issues such as socio-economic, racism, and sexism, place African American women at particular risk for substance abuse. Limited availability to treatment, a lack of appropriate treatment and poor social supports impact recovery and places these women at risk for relapse. This article provides an overview of the current literature regarding substance abuse, treatment and recovery in African American women. PMID- 11855010 TI - Hmong and prenatal care. AB - Providing prenatal care to the Hmong can be a challenge for the Advanced Practice Nurse. Although some Hmong have become acclimated to Western medicine, others may not understand Western preventative medicine, health promotion or prenatal care. Traditional cultural beliefs and socioeconomic status may contribute to the barriers a practitioner faces when providing prenatal care to a Hmong client. Practitioners must assess each client and identify barriers. Integrating cultural beliefs and Western medicine practices can then foster prenatal care. PMID- 11855011 TI - Comparison of multicultural literature in three health professions: physical therapy, nursing & social work. AB - All health professions' education programs are mandated by their accrediting agencies to include diversity and multicultural content within their curricula. This article reviews literature from the three professions of nursing, social work, and physical therapy that indicates that they accomplish this at different places on the continuum of curricular integration and transformation as identified by Banks. An examination of the complexity of each profession's standards, and a comparison of the depth of cultural competency within each profession is included. Even with the more 'experienced' professions of social work and nursing, there appear to continue to be inconsistencies in the structuring and teaching of multicultural content. PMID- 11855012 TI - An evidence based approach to a perennial problem: pressure ulcers. AB - The nursing care required to prevent the development of pressure ulcers on patients of various ages and with a range of physical and medical problems is an increasing challenge for nursing staff. The challenge for the nursing staff is multifactorial. Concerns include a complexity of factors associated with discerning, interpreting and implementing the results from evidence based studies, and maintaining cognisance of the ever-spiralling health cost while balancing this with the patient's wishes. The prevention of pressure ulcers is a cost-effective strategy about which little is written. This paper defines pressure ulcers using the International Classification for Nursing Practice (International Council of Nurses, 1999), and outlines the evidence featured in the research literature on the strategies for prevention. PMID- 11855013 TI - Exploring the experiences of beginning registered nurses entering the acute care setting. AB - This study begins to explore some of the social and political issues surrounding the practices of the graduate nurse. Utilising an ethnographic methodology with a critical intent, 4 graduate nurses describe their experiences of clinical practice. The major themes raised or issues that were embedded within the nurses' stories revolved around power and control enmeshed within nursing practice. The themes discussed relate to the graduates' perception of their own competence, and the concepts of the ideal nurse, the socialisation of graduates into the ward culture, being on insider or outsider and a good or bad nurse. The resulting discussion utilises the theoretical framework of Foucault's governmentality to suggest ways in which nurses and graduates might make sense of these issues. PMID- 11855014 TI - Promoting self-help strategies by sharing the lived experience of arthritis. AB - A qualitative approach informed by the phenomenological concept of lived experience using semi-structured interviews explored the experience of living with arthritis. Audio-taped transcribed conversations were analysed using a computer-assisted thematic analysis procedure. Strategies were offered for managing mornings, ensuring personal comfort, keeping a positive attitude, doing housework, cooking and meals, getting exercise, existing in day-to-day life, living at a slower pace, acknowledging feelings, dealing with depression, trying alternative treatments, accepting illness, getting sleep and rest, getting help, using help and handy gadgets, having emotional support, having determination, managing pain relief using distractions, making adjustments, planning ahead, maintaining independence, having a social life, managing stress, adapting around young children, and facilitating self-awareness. PMID- 11855015 TI - Coping with crisis: how Australian families search for and select an aged care facility for a family member upon discharge from an acute care setting. AB - Searching for, and selecting, an aged care facility for a family member upon his or her discharge from an acute setting is known, anecdotally, to be a challenge; but the process itself, and its effects on families, has seldom been examined. In this exploratory/descriptive study, face-to-face in-depth interviews were conducted with sponsors (often family members) of residents who had been admitted to an aged care facility within the past 90 days, following their discharge from an acute care setting. Where possible, both sponsor and resident were interviewed. This paper reports on the participants' perceptions of the search and selection process. Specific areas of interest were probed: Why and how the decision was made to seek residential placement; when and how the search for an aged care facility began; factors that were important in the family's final selection process; what happened when families, either initially or in the long term, were unable to find a place in an aged care facility; and the family's perspective about the efficacy of the search and selection process and its effect on the well being of the family. Residents are often passive in the search and selection process, while sponsors are often actively involved. Very few residents or sponsors consider planning for an aged care facility prior to hospitalisation, and there is often the perception by families of having very little support at this stressful and emotional time. The decision to relocate a family member in these circumstances is a stressful experience and should be viewed as a family crisis, particularly if unexpected such as upon discharge from an acute setting. These findings provide valuable insights that can be used to guide and assist families experiencing this process, as well as health care professionals working with families in this situation. PMID- 11855016 TI - Bullying nurses at work: theorizing a gendered experience. AB - This paper is about bullying among nurses at work. It presents the psychoanalytically based theory that workplace bullies, impaired during infancy by primary caregivers who were less than loving, project their own hostile personalities onto others and then relate to others without empathy or understanding, in demeaning ways. When these hostile people are employed in a masculine workplace, they protest against the gendered imperatives imposed upon them, hysterically. Because of the masculinization of the workplace, hysterical bullying varies according to gender, with women bullying in a hostile connected way, and men bullying in a hostile separated way. Research data gathered in Canada in the 1990s is utilised in presenting the theories. Suggestions about anti-bullying practices that arise from this theoretical analysis conclude the work. PMID- 11855017 TI - Evaluating a problem based learning course: an action research study. AB - Problem based learning (PBL) has been widely used in the United States, United Kingdom and Australasia in undergraduate nursing education to develop critical thinking and problem solving skills. PBL has been used since 1996 in a Bachelor of Nursing course at a New Zealand tertiary institution, and several modifications have been made to foster effective learning. The 'pure' PBL process has been adapted to move students gradually from teacher direction to taking responsibility for their learning. This has provided the opportunity for students to develop critical thinking, problem solving, information retrieval and evaluation skills, and group process skills over an 18-week period. Because rigorous evaluation of these changes had not been formally undertaken, the purpose of this study was to evaluate how the current format was developing students' understanding and integration of knowledge. Two cycles of the action research method (Cardno and Piggot-Irvine, 1994) were used, involving 4 lecturers and 17 students. Data was collected both quantitatively and qualitatively over a 16-week period. Findings indicated the importance of: explaining the purpose and process of PBL; communicating in detail the role of both students and lecturers; keeping communication lines open; addressing timetabling issues and valuing this method of learning for nursing practice. Implications for nursing education are addressed. PMID- 11855018 TI - Student learning through video assessment. AB - This paper reports on the use of video and self-assessment as a tool to guide and support undergraduate students in the practice of nursing. This method of learning and assessment promoted self- and peer evaluation as well as critical thinking. It encouraged students to take more responsibility for their learning and assisted academic staff and students to identify learning issues associated with clinical practices. Evaluative comments by teachers, students and educational experts have established the credibility and continued use of this approach within the Bachelor of Nursing program at a major university in Sydney, Australia. PMID- 11855020 TI - Setting the agenda for consumer-oriented care in the new millennium. PMID- 11855019 TI - Nursing skill assessment within populations: scale development and testing. AB - This study examined the reliability and validity of an instrument to measure the skill level of registered nurses within an Australian state. Internal consistency and construct validity of the scale and subscales were investigated in a sample of 1223 registered nurses from varying practice areas. The scale consisted of 105 skills within 13 practice domains: nursing practice, professional practice, acute care, community and public health, rural and remote area nursing, service delivery, self-management, supervision, resource management, improving processes, training and development, research, and technology. Twelve subscales obtained reliability co-efficients of 0.85 or more, with one at 0.81. The 13 factors explained 64.5 per cent of the cumulative variance, adequately capturing the concept of nursing skill. This scale, and its subscales, represent valid and reliable tools for the assessment and monitoring of skill levels in a population of nurses, within some limitation. PMID- 11855021 TI - Exemplar. AB - As I gain more experience as a practitioner, I find I am increasingly able to articulate my personal philosophy of nursing. I have learned that, as a professional nurse, my personal values and philosophies of nursing do affect the nature of the care I deliver and the quality of relationships I form within my daily practice. Becoming involved in an incident like the following has enabled me to develop an understanding of complex interpersonal and interprofessional situations. Unspoken grief, the devastating death of a child and the behaviour that surrounds such an event are part of what makes my experience with Kate and her parents so important to me. By reflecting on this incident, I have developed a new insight and have found new meanings from my experience. This has further allowed me to uncover and explore the implications this might have in my future practice. PMID- 11855022 TI - Literature review: the reluctance for the use of opioid analgesia in the client with cancer. AB - Many articles have been written and studies conducted into why our patients' pain is not being managed more effectively. The myths and fears still persist, yet the solution is known. Why, then, can't our profession help those we care most about- our clients? PMID- 11855024 TI - Withdrawal of treatment in the intensive care unit: the ethical and legal issues. AB - Many articles have been written and studies conducted into why our patients' pain is not being managed more effectively. The myths and fears still persist, yet the solution is known. Why, then, can't our profession help those we care most about- our clients? Intensive care units (ICUs) have, over the last 30 years, become places of life-saving miracles, and as such, moral dilemmas have also been created. As early as 1976, applications have been made to courts for the withdrawal of life-sustaining treatment (Wallace, 1995). The decision to withdraw treatment is one faced by health professionals every day in the ICU, and there are many legal and ethical issues to consider. The emotional, and other, stakes are also high in this area for patient, family and care team. This paper discusses ethical and legal issues highlighting the principles, doctrines, legislation and precedents especially important in the making of the decisions to withdraw treatment. An ethical framework grid is suggested to assist with the decision-making process. PMID- 11855023 TI - Health assessment and its relationship to nursing practice in New Zealand. AB - Health assessment has been an integral component of nursing education in Australia for over a decade. New Zealand has only recently embarked down this path and might benefit from the Australian experience. This article will discuss health assessment in the context of three issues currently topical in nursing in New Zealand. The issues are annual registration based on evidence of competence to practice, a review of undergraduate curricula, and the development of nurse practitioner/advanced nurse practitioner roles. The meaning of the concept 'health assessment' is also clarified in order to provide consistency as new initiatives in nursing are currently being developed. PMID- 11855025 TI - Have you got an attitude problem? Caring for illicit drug-using patients. AB - The use of illicit drugs in an environment of prohibition results in major health risks to the drug-using population. Substantial morbidity and mortality in this group is identified as a priority area for public health action. Undergraduate nursing programs prepare students to manage clinical emergencies resulting from drug use, to undertake health promotion activities in a harm minimization policy environment, and to provide appropriate community referral. It is a cause for concern, therefore, that a group of senior undergraduate nursing students express negative perceptions of a drug-using individual's personality and behaviour. It is imperative that undergraduate nurse educators find ways to challenge student attitudes from the first semester of their program. Nurses in the clinical setting must display professional values that emphasize the dignity and worth of all individuals. They have an opportunity to make a difference to the health outcomes of this marginalized group of health consumers. PMID- 11855026 TI - Effective communication in an intensive care setting: nurses' stories. AB - Effective communication has been long considered an important aspect of nurse patient interactions. However, follow up of people who have been patients in intensive care units (ICUs) indicates that nurses' communication in this setting is at times unsatisfactory. Patients have described how they were left feeling frustrated and alienated by this failure in the communication process and unconscious patients report how they were aware of nurses' attempts at communication with them while unconscious (Lawrence, 1995). Because most of the research to date has focused on patients' perceptions of nurses' communication in an intensive care unit, this study sought to explore what nurses believed constituted effective communication in an intensive care setting. Therefore, a qualitative study was undertaken to explore nurses' perceptions of effective communication with patients in an intensive care setting. The stories of four intensive care nurses were utilized as the means of data collection. The analysed data revealed the following themes: nurses' perceptions, presencing and reassurance. The findings have relevance for nurses in many settings. PMID- 11855027 TI - In the nursing interest: reading and thinking critically in the new millennium. PMID- 11855028 TI - Nurses as social control agents? AB - Social control is a phrase that conjures images of oppression in the minds of most. Nurses have limited control within the healthcare arena, and many would argue that nurses are angels of advocacy, rather than agents of social control. However, nurses do have a place in social control, particularly to socialize nurses and patients into their roles, and to promote society's view of health. PMID- 11855029 TI - Academic and clinical collaboration. AB - Australian nurses have been slow to embrace and subsequently publish research that has been achieved through collaboration between academic and clinical nurses. If nursing is to continue its push towards greater professionalism, nurses from both areas must recognise the importance of collaboration in producing meaningful high quality research. Collaboration between nurse academics and clinical nurses has been advocated as a means of ensuring clinically relevant research. This paper explores collaboration and the issues that impinge on nurses conducting such research. PMID- 11855030 TI - Testis unis, testis nullas: one witness is no witness. Criticisms of the use of oral history in nursing research. AB - In research, there is no perfection: no perfect method, no perfect sample, and no perfect data analyses tool. Coming to this understanding helps the researcher identify the inadequacies of their preferred method. This paper discusses the criticisms of the oral history method, drawing reference to its challenges and difficulties in relation to its use in nursing research. Oral history has the advantage over more traditional historical approaches in that the narrators can interpret events, personalities and relationships within the interview that are not accessible from written sources. The oral history interview may also provide a forum for unveiling documents and photographs, which might not have been otherwise discovered. Nonetheless, oral history, like most methodologies, is not flawless. This paper discusses the limitations of oral history and suggests ways in which a nurse can use oral history to provide an account of aspects of nursing history. PMID- 11855031 TI - Hidden treasure: nurses writing for publication. AB - Nurses have a wealth of tacit and explicit knowledge, the value of which remains hidden because of a fear of authorship. How can nurses overcome their intimidation by the pen? In this paper, the personal and intrinsic are integrated with the public and literary as key issues on nurses' authorship are explored. Motivation and disincentives are balanced against helpful strategies for new writers such as cultivating a positive attitude, linking with a critical colleague, consulting 'how to' books, and journal keeping. In addition, originality, creativity, ethics and the shadow side of publishing are discussed. PMID- 11855032 TI - The seminar: how to develop and deliver a dynamic presentation. AB - Oral presentation of information and facilitation of discussion by students through seminar presentation are now major components of many graduate nursing courses. Facilitators of seminars need to use a wide variety of techniques to structure the interaction of learners to develop higher order cognitive skills. Seminars are a method of motivating and involving participants through discussion and interaction with facilitators and peers. Participants build on each others' knowledge and ideas through discussion while also discovering new knowledge for themselves which generates and sustains interest, attention and critical thinking. Students' previous experiences in communicating information and ideas, however, have been focused primarily on writing reports and essays, not in oral presentation that often leaves graduate students at a loss to know how to develop and deliver effective and interesting seminars. The authors have identified a lack of easily accessible literature for nurses on how to conduct seminar presentations. The purpose of this paper, therefore, is to offer some practical suggestions with respect to the development and delivery of successful seminar presentations. The authors will discuss key skills such as how to prepare and structure seminars, how to facilitate discussion, use visual aids and how to deal with problems common to the conduct of seminars. PMID- 11855033 TI - The impact on attitudes towards cultural difference of participation in a health focused study abroad program. AB - The changes in attitudes towards cultural difference of seventeen participants in a three-week community health study abroad program to Nepal were compared with the changes in attitudes of a similar group who did not participate in the tour. Participants in the tour group were surveyed eight weeks prior to departure and in the last week of the tour using a twenty-six item questionnaire employing a six-point forced-choice response scale. The responses of participants in the tour group showed significant shifts in relation to eight items compared while the responses for the control group showed no significant shifts. Observed student advantages of participation in this study tour included the development of independent behaviour and positive cultural adjustment and adaptation. PMID- 11855034 TI - Changing the culture in nursing--a strategy. PMID- 11855036 TI - Improving nursing care through mentored research: a paper within a paper. AB - In 1995 Western Sydney Area Health Service (WSAHS) and University of Western Sydney Nepean (UWSN) entered a strategic alliance to develop a nursing research culture in Western Sydney. As a result, the WSAHS/UWSN Professorial Nursing Unit (PNU) was established in 1996: Its mandate was to develop a variety of strategies to promote a nursing research culture in the area. One of these strategies was to encourage mentored research projects between personnel in the PNU and inexperienced clinical nurses. This paper describes one such project. In addition, it contextualizes the study in relevant literature relating to mentored research and outlines both the issues and the benefits associated with such projects. PMID- 11855035 TI - Medicalisation of life: are nurses involved? PMID- 11855037 TI - Patient satisfaction: expectations and experiences of nursing care. AB - This qualitative descriptive study aimed to evaluate surgical patient satisfaction with nursing care. A convenience sample of six general surgical patients participated in semi-structured interviews. Satisfaction was evaluated by comparing patient expectations with actual experience of nursing care. Data analysis revealed patient satisfaction with the personal and professional nature of nurses. Participants understood information from nurses and were satisfied with nursing ability. Participants were dissatisfied with minimal nurse contact, lack of personalized care and lack of information about the operation, recuperation and minor treatment options. The challenge for nurses will be to further investigate and address these deficiencies to improve satisfaction with care, thus contributing to better health outcomes. PMID- 11855038 TI - 'Appears to have slept well': assessing sleep in an acute care setting. AB - Sleep disturbance is a common occurrence in the hospital environment where the patients' perceptions of their own sleep are rarely considered in the planning of care. This study aims to determine the difference between subjective (Verran/Snyder-Halpern Sleep Scale), objective (Patient's Sleep Behaviour Observational Tool) and physiological (Actigraph) assessments of a patients' single sleep event within an acute care setting by examining the variables Total Sleep, Midsleep Awakening and Sleep Latency. Results indicate no statistically significant difference between instruments for the assessment of total sleep. Midsleep awakening was of questionable value when assessed in the clinical environment. Sleep latency could be useful following further attention to defining the length and time of onset of the nurse observation period. PMID- 11855039 TI - Getting it right: appropriate therapeutic recreation programs for community based consumers of mental health services. AB - Over the last two decades in Australia, the deinstitutionalization process, which began with the intent of moving consumers of mental health services from in patient facilities and then seeking to integrate these same individuals into the community, has served to highlight a wide range of consumer needs that have remained largely unfulfilled throughout the process. One such need has been the provision of appropriate therapeutic recreation programs for the community based consumers of the various state co-ordinated mental health services. This paper argues a case for a change in the approach which professional staff provide and lead therapeutic recreation based programs to enable participants to be empowered, rather than disempowered, through their involvement. Further, this paper contends that there is a need for health care staff, more generally, to accept the concept of such programs for the community based consumers of various mental health services as a valued one. PMID- 11855040 TI - Ethics, the health assessment interview, and the older patient. AB - As a requirement for the first year nursing subject 'Health Assessment' at Flinders University of South Australia, the task was given of examining Ethics and how this relates to the health assessment interview. Due to the negative stereotypes of older people and the subsequent treatment of the older patient, focus is placed on this particular age group. A definition of Ethics is given, with further discussion relating specifically to autonomy, respect and justice. This paper is not intended to be a major dissertation on Ethics and the Four Principles, because at the time of writing this the author had not studied ethics. Instead, it is hoped that other nurses and nursing students may learn as much about the difficulties of the older patient as the author has herself, and examine possible prejudices towards patients of any age, particularly the vulnerable older patient. PMID- 11855041 TI - Quality medication administration. AB - Quality medication administration is not simply a matter of adhering to the 'five rights'. The nurses' role in medication management is influenced by a multitude of factors not always within their control. Yet nurses maintain a duty of care to uphold safe and appropriate nursing interventions in pursuit of quality drug use. Whilst legislation and nursing standards offer principles and parameters for such interventions they can not provide directives. Hence, it remains each nurse's responsibility to understand the factors that influence medication management, to attain best practice on the patient's behalf. PMID- 11855042 TI - Restraints must be a last resort! PMID- 11855043 TI - Family responses to disclosure of self-as-lesbian. AB - Disclosure or nondisclosure of self-as-lesbian to family is a difficult task. Eleven lesbians aged 25-47 years old described their experiences of disclosure or nondisclosure to family. The primary source of data collection was focus groups. Family disclosure was preceded by recognition of self-as-lesbian. Five story patterns of lesbians and their families related to disclosure or nondisclosure were derived from text analysis. The participants also described missing sociocultural landmarks and symbols for lesbian lives. Implications are included for lesbians who are contemplating disclosure and their health care providers. PMID- 11855044 TI - A comparison of two groups of women with bulimia nervosa on symptom onset. AB - Bulimia nervosa (BN) is a significant mental health problem for women. Both children and adolescents develop risk factors for BN that are identifiable. An important role for psychiatric-mental health nurses is the prevention of psychiatric disorders through risk factor identification and intervention for early symptoms. The purpose of this study was to extend recent research on the sequencing of risk factors and symptoms in the development of bulimia nervosa. Symptom history was collected on 71 women who agreed to participate in research on eating disorders (EDs). Women were divided into two groups based on whether their first symptom manifested was bingeing (BF) or dieting (DF). The groups were compared on age of symptom onset, weight history, and history of diagnosis. There were significant differences between the groups, and the present findings contradict some of the important conclusions from three previous studies. The results are discussed and implications for strategies for early detection and prevention are presented. PMID- 11855045 TI - Comparison of the stressors and coping behaviors of homeless, previously homeless, and never homeless poor children. AB - The purpose of this study was to compare the stressors and coping behaviors of homeless, previously homeless, and never homeless poor school-aged children. Content analyses of interview data from 132 children were used. Forty-four (68%) of the homeless children identified at least one stressor related to being homeless. The three groups of children were similar in regard to stressors related to the family, self, peers, health, school, and environment. In regard to the violent behavior category, significantly more of the previously homeless and never homeless children expressed stressors compared to the homeless children. Significantly more previously homeless and never homeless children identified more social support coping behaviors compared to the homeless children. The three groups of children did not differ in regard to the use of behavioral and cognitive coping strategies. PMID- 11855046 TI - Patterns of depressive symptoms in three groups of depressed adults. AB - This study examined patterns of depressive symptoms experienced by acutely depressed inpatients, previously hospitalized depressed outpatients, and depressed outpatients who had never been hospitalized for depression. The groups' symptom profiles were compared to determine whether the 21 major depressive symptoms measured by the Beck Depression Inventory were similar or different for the three groups. The groups differed significantly on five depressive symptoms that are classified as affective/cognitive symptoms (sadness, guilt, self-blame, indecisiveness, suicidal ideas) and one that is considered a somatic/vegetative symptom of depression (anorexia). Thus, commonly used diagnostic criteria may not reflect the full range or temporal patterning of symptoms experienced by depressed persons at varying levels of acuity or severity and in different treatment settings. PMID- 11855047 TI - Assessment of cardiovascular risk factors among residents of a city in Jordan. AB - Cardiovascular diseases are a leading cause of morbidity and mortality in many countries. The purpose of this study was to assess cardiovascular risk factors in a stratified randomly selected sample of a city near Amman, Jordan. A stratified sample of two hundred and nine randomly selected households were selected for this study. Adults from each of the households who agreed to participate in this study were asked about their cardiovascular risk factors including cigarette smoking, high blood pressure, cholesterol, diabetes, obesity, and sedentary lifestyle. The sample consisted of 84 males and 125 females ranging in age from 17 to 93 years with a mean age of 37 years. Findings identified significant cardiovascular risk factors included cigarette smoking, obesity, hypertension, stress, and diabetes. Cigarette smoking was much more common in men than women. Implications for nurses are discussed with suggestions for future research. PMID- 11855048 TI - Secondary prevention of breast cancer in female Japanese citizens residing in the USA. AB - The purpose of this paper is to advise medical care providers in the USA about appropriate secondary prevention approaches for breast cancer in female Japanese citizens residing in the USA. The Japanese population is increasing yearly in the USA. It is estimated that 2,000 Japanese nationals reside in just the Cincinnati metropolitan area. It is important to understand the impact and health risks of breast cancer for this population, as well as identify cultural behaviors and language difficulties. Clinical implications for the secondary prevention for breast cancer in this population are discussed. PMID- 11855049 TI - The effects of cooperative learning on students enrolled in a level 1 medical surgical nursing course. AB - This study was undertaken to create an environment that the literature contends will be more conducive to learning and one that will build the necessary social skills impacting students' performance. These social skills are essential if a nurse is to be effective in her role as a competent provider of care as well as an effective member of the inter-disciplinary health care team. The concept of cooperative learning was applied to nursing theoretical content in an effort to (a) decrease attrition rates in a first level medical-surgical nursing course, (b) increase student's knowledge and (c) enhance student's patient-teaching skills. Increased knowledge was assessed by student's performance on teacher-made paper and pencil examinations. The Classroom Life Instrument questionnaire was used for feedback on the teaching strategy and the Griffin tool was the basis for the pre-test and post-test assessment and implementation of the patient teaching plan. PMID- 11855050 TI - Evaluation of an educational intervention to increase cultural competence among registered nurses. AB - The purpose of this two group intervention study (N = 94) was to determine if RNs who participated in "culture school" improved levels of cultural competence to a greater extent than RNs who attended nursing informatics classes. The Giger and Davidhizar Transcultural Assessment Model/Theory (GDTAMT) was the study's theoretical foundation (Giger & Davidhizar, 1995). A sample of 94 participants, was identified from a randomized group of all Jefferson County, Alabama RNs. Randomly assigned participants (stratified by race) experienced 8.5 hours of either culture school or nursing informatics classes and completed survey tools in three phases (pre-intervention, immediate post intervention, three week follow up). The Cultural Self-Efficacy Scale (CSES) by Bernal and Froman (1987), knowledge base questions by Rooda (1990), and demographic profiles were used. Concepts empirically measured using these tools were analyzed by transcultural nursing experts for their congruence with GDTAMT. Using repeated measures analyses of convariance (race), significant differences between groups for both scales were found. Culture school participants demonstrated significantly more cultural self-efficacy and cultural knowledge, and these improvements remained during phase three. Further research is recommended to allow for greater generalizability of findings, an examination of client perceptions, and actual nurse behaviors. PMID- 11855051 TI - [Criteria for adequate respiratory support during transfer of patients to spontaneous respiration]. AB - Time course of circulation, oxygen transport and consumption parameters were studied in patients with parenchymatous pulmonary diseases during their transfer to spontaneous respiration under conditions of pressure support ventilation after long forced ventilation of the lungs. The oxygen cost of respiration can serve as a reliable criterion of respiratory support adequacy when the use of a respirator is discontinued. With oxygen cost of respiration at least 14%, a decrease of respiratory support is hardly possible without decompensation of the respiration system and circulation, which dictates monitoring of this parameter during transfer of patients to spontaneous respiration. PMID- 11855052 TI - [Acute lung injury syndrome and hemodiafiltration]. AB - Acute lung injury syndrome is one of the most frequent components of polyorgan failure, particularly resultant from sepsis. Replacing renal therapy can correct uremic disorders in the hemostasis and gaseous exchange disorders in the lungs by eliminating excessive fluid in case of hyperhydration and by eliminating inflammation mediators. The present study demonstrated a favorable impact of hemodiafiltration on ventilation oxygen status of patients with acute lung injuries, which was confirmed by positive changes in blood oxygen pressure, intrapulmonary shunt, alveolar-arterial oxygen difference, etc. Changes in oxygen transport corresponded to those in patients with sepsis and shock without hemodiafiltration. PMID- 11855053 TI - [Effect of perftoran on post-resuscitation recovery of the central nervous system]. AB - The effect of perfluorane on survival and restorative process in the brain were studied in rats subjected to 12-min arrest of systemic circulation. Perfluorane in a single dose of 5-10 ml/kg was injected intraperitoneally 30 min after the beginning of reanimation. The drug did not affect the postreanimation death of animals and time course of neurologic deficiency disappearance. Perfluorane activated behavioral reactions and prevented development of dystrophic changes in the brain structures of rats highly sensitive to hypoxia. PMID- 11855054 TI - [Effect of perftoran on morphology and rheologic properties of erythrocytes in patients with severe trauma and blood loss]. AB - Erythrocyte morphology and function, plasma viscosity, central hemodynamics and oxygen status were studied in patients with severe injury and blood loss. The percentage of morphologically changed erythrocytes (mainly stomatocytes) increased, their filterability changed negligibly. Changes in these parameters were studied during intravenous infusion of perfluorane. Perfluorane therapy during the acute period of severe injury decreased total peripheral and pulmonary vascular resistance and reduced the entry of an extra amount of structurally changed erythrocytes from peripheral tissues into the blood stream. PMID- 11855055 TI - [Experience in using perftoran in combined treatment of placental insufficiency in severe gestosis]. AB - Twenty-two women with severe gestosis were examined during weeks 27-38. They were divided into 2 groups with different intensive care protocols: 1) osmooncotherapy and 2) 3-4 infusions of perfluorane in a dose of 3-4 ml/kg every other day. Addition of perfluorane to combined therapy promoted a more rapid stabilization of hemodynamics, metabolism, and helped prolong the pregnancy to 37-38 weeks. PMID- 11855056 TI - [Treatment of DIC syndrome in patients with severe craniocerebral trauma]. AB - Transfusion of fresh-frozen plasma with fraxiparin is the most rational basic therapy for the first and third clinical pathogenetic variants of subacute disseminated intravascular coagulation in multiple-modality treatment of severe craniocerebral injury. In the second variant of this condition, high doses of proteinase inhibitors in combination with minimum heparin doses are the most rational. PMID- 11855057 TI - [Status of cerebral perfusion pressure and cerebral oxygenation in the acute period after cranio-cerebral injury]. AB - Sixty-one patients with severe craniocerebral injuries were examined on days 1-3 after the injury. Consciousness was no more than 8 points by the Glasgow Coma Scale. The patients were divided into 2 groups: 1) favorable outcomes and 2) unfavorable outcomes. Gaseous composition of arterial and venous blood was analyzed in all patients, intracranial pressure (ICP) and cerebral perfusion pressure (CPP) were measured, artificial ventilation of the lungs (AVL) and noninvasive monitoring of cerebral oxygenation (CO) were carried out. The results indicate that continuous monitoring of CPP and CO helps control the time course of cerebral blood flow, thus promoting early detection and effective treatment of cerebral ischemia in critical patients. PMID- 11855058 TI - [Role of disrupting the antioxidant status of the body in forming the endogenous intoxication syndrome in patients with toxic and terminal stages of peritonitis]. AB - Fifty-one patients with diffuse peritonitis were treated. 40 of these presented with the toxic stage and 11 had terminal stage with lethal outcomes. Studies of the antioxidant status showed an important role of lipid peroxidation metabolites in the formation of endogenous intoxication during the early postoperative period, manifesting, among other things, by decreased albumin transporting function. Analysis of correlations showed some relationships in the studied parameters which can be used in complex with other values in clinical laboratory diagnosis of endotoxicosis severity in peritonitis and in evaluation of the time course of this condition during intensive care. It can serve as a prognostic criterion of peritonitis development and outcome. PMID- 11855059 TI - [Dynamics of indicators of blood coagulation and fibrinolysis in patients with diffuse peritonitis]. AB - Blood coagulation and fibrinolysis were studied in 34 patients with diffuse peritonitis treated in intensive care wards of common profile. Forced ventilation of the lungs was carried out for 2-26 days in 32 of 34 patients. The causes of peritonitis were perforating gastroduodenal ulcers (22 cases), acute perforating ulcers of the small intestine (5 cases), and acute phlegmonous perforative cholecystitis (7 cases). Disseminated intravascular coagulation, depressed fibrinolysis first in the plasma and then in whole blood were observed early (2 16 h) after surgery in all patients (17 survived and 17 died). These changes progressed by days 3-5 of treatment in all patients, being the most pronounced in cases with complications (failure of sutures after gastroduodenal ulcer suturing, acute gastroduodenal ulcers complicated by hemorrhages, acute perforating ulcers of the small intestine, etc.), particularly in the patients who died. PMID- 11855060 TI - [Use of computer monitoring in assessing hemodynamic reactions in hypertension patients during endoscopic gynecologic surgery]. PMID- 11855062 TI - [Comparative evaluation of methods of analgesia after operations on the stomach]. AB - A general somatic syndromal approach to evaluation of clinical status by SAPS II and APACH II scores was used in 41 patients operated on for gastric cancer. Epidural analgesia in these patients promoted rapid regression of the severity of condition, did not require high opioid doses, had a positive effect on the peristaltic activity of the intestine, and accelerated treatment in intensive care wards. PMID- 11855061 TI - [Indicators of lipid metabolism in patients in critical condition]. AB - Plasma lipid metabolism was studied in critical patients with severe combined injuries. Decrease in cholesterol concentration and increase in triglyceride concentrations were more pronounced in patients with lethal outcomes. The authors conclude that plasma cholesterol and triglyceride concentrations can be used as prognostic signs in patients with severe combined injuries. PMID- 11855063 TI - [Use of lornoxicam for analgesia in the early postoperative period]. AB - Lornoxicam was used for analgesia in 64 patients on days 1-2 after extensive interventions. The drug efficiency and safety were evaluated depending on the dose and route of administration. Intravenous infusion of lornoxicam in a daily dose of 24 mg (basic therapy) did not involve the use of opioids in 35% patients and its analgesic effect was higher than that of promedol monotherapy. Combined therapy with lornoxicam and promedol allows reduction of promedol dose by 25-50% and the incidence of untoward effects by 27-44%. PMID- 11855064 TI - [Neurobiological mechanisms of massive blood loss]. AB - Molecular mechanisms of neuronal death (apoptosis and necrobiosis) in massive blood loss and at various stages of the postreanimation period are investigated from the neurobiological viewpoint. The activities of these processes are quantitatively evaluated and their specific features in incomplete cerebral ischemia, reperfusion (reanimation), and during the postischemic (postreanimation) period are analyzed. PMID- 11855065 TI - [Effect of occlusive cerebral ischemia on functional status of the internal organs]. AB - Biochemical parameters characterizing visceral functions were measured in the plasma of rats with brain ischemia induced by double occlusion of the carotid arteries. Functional insufficiency of the viscera is gradually forming in the course of occlusive ischemia of the brain. Functional insufficiency of the liver was observed in animals with severe neurological deficiency and subsequent lethal outcome. In rats resistant to ischemia, visceral dysfunctions develop in a certain succession, starting from the pancreas and followed by the myocardium, liver, and kidneys. PMID- 11855066 TI - [Features of hormonal status in men during long-term coma]. AB - Serum hormonal profiles were studied in 48 patients during long (6-56 days) coma. Low T3 and T4 and an essential increase in the levels of hydrocortisone, prolactin, estradiol, and particularly progesterone (20-fold) in the presence of normal LH concentration and decreased level of FSH were observed on days 7-16. Changes in the concentrations of sex hormones, similar to the "false pregnancy" condition, were the most pronounced in the patients who later died. PMID- 11855067 TI - [Effect of laser exposure on hemostatic parameters in the preagonal and post resuscitation period]. AB - Effects of intravascular low-intensive laser exposure of the blood on the hemostasis during acute blood loss and the early postresuscitation period after 4 min clinical death were studied on narcotized dogs (8-17 kg) of both sexes with different initial levels of heparin. During the preagonal period laser exposure caused hypercoagulation in animals with initial heparin content below 60 micrograms/ml. This acceleration of blood clotting prevented a drop in the activity of antithrombin III and hypercoagulation by the third hour of postresuscitation period. In animals with initial heparin content more than 60 micrograms/ml laser exposure caused hypocoagulation in the presence of reduced activity of antithrombin III by the third hour of the resuscitation period. PMID- 11855068 TI - [Immune status in the perioperative period in gestosis]. AB - Search for new methods for the earliest possible diagnosis of pre-eclampsia for timely purposeful correction of the consequences of developing disorders remains the central problem of obstetrics. We evaluated the viability of some organs by measuring specific antibody titers. The study was carried out in healthy women and patients with severe gestosis. Gestosis induced pronounced degenerative processes involving predominantly the liver, kidneys, myocardium, lungs, and ocular retina. This method early detects organ changes in gestosis and thus allows timely specific therapy. PMID- 11855069 TI - [Clinical-radiological and laboratory parallels in respiratory distress syndrome in patients with gestosis]. AB - Clinical and laboratory studies were carried out in 125 pregnant patients with gestosis of different severity. Respiratory distress syndrome was diagnosed, its severity depending on the severity of patient's status. Platelet changes indicated involvement of the lung parenchyma and can serve as an additional laboratory criterion in the diagnosis of the respiratory distress syndrome in pregnant patients with gestosis. PMID- 11855070 TI - [Liquid pulmonary ventilation, its possibilities and prospects (current state of the question)]. PMID- 11855071 TI - [Use of position tests for detecting oxygen status disorders and correction of them in patients with severe blunt chest injury]. AB - Relationship between alteration of body position and parameters of central hemodynamics and oxygen status were studied in patients with severe blunt chest injury. Use of position tests in this patient population helped detect disorders in the central hemodynamics, oxygen transport and consumption. Argument increase of volumetric loading promoted an increase in oxygen delivery. PMID- 11855072 TI - [Pathogenetic principles of renal replacement therapy in resuscitated patients]. PMID- 11855073 TI - [Oxygen debt as a prognostic criteria in patients with multiple organ failure]. AB - Signs of oxygen debt were studied in 28 patients with systemic inflammatory response and polyorgan failure. The parameters of oxygen debt, oxygen consumption, saturation of mixed venous blood, and heart rate can be used as criteria of therapy efficiency. The probability of a favorable outcome is increased if the mean 24-h oxygen debt is less than 30 ml/min x m2, oxygen consumption more than 158 ml/min x m2, mixed venous blood saturation less than 64%, and heart rate less than 114 stroke/min. PMID- 11855074 TI - [Comparison of effectiveness of BCG vaccination and preventive therapy in Japanese settings, with special emphasis on the sensitivity and specificity of tuberculin testing]. AB - BACKGROUND: BCG vaccination in low prevalence countries is controversial. Most discussions have been done by the comparison of benefit, side effect and cost of BCG vaccination. No discussion has been done on the disadvantage of BCG vaccination from the point of view of loss of sensitivity and specificity of tuberculin test on the diagnosis of LTBI. METHOD: Three groups, i.e. the BCG vaccination group with preventive therapy under worsened sensitivity and specificity of tuberculin test due to previous BCG, no intervention group and non BCG vaccination group with preventive therapy under standard sensitivity and specificity of tuberculin test were set up. The target population was a cohort of Japanese who are born at the year with 0.1% annual risk of tuberculous infection. The TB incidence, TB related mortality (including death by BCG and preventive therapy), TB related loss of DALY (disability adjusted life years), and direct medical cost of the above three groups, for the cohort only and the cohort including secondary TB cases from the clinical cases in the cohort. RESULTS: Under the current program conditions, the merit of BCG vaccination is greater than the merit obtained from the preventive therapy without BCG. Although the medical direct cost is lowest among the preventive therapy group without BCG, next without BCG or preventive therapy and highest among BCG group. Under BCG group, too extensive screening for LTBI does not reduce the loss of TB related DALY, whereas in the non BCG group, extensive screening will help to reduce the loss of TB related DALY and if more extensive screening can be done, the loss of TB related DALY in the non BCG group can be less than that in the BCG group. CONCLUSION: At present extensiveness of screening for LTBI and BCG vaccination contribute to the reduction of TB and loss of TB related DALY. Possible extensive screening without BCG may be able to reduce loss of TB related DALY in comparison to the BCG vaccination group. PMID- 11855075 TI - [Clinical investigation of pulmonary Mycobacterium kansasii infection in our hospital]. AB - There is evidence that the number of non-tuberculous mycobacterium (NTM) cases is increasing at least in some areas of the world and as possible causes of the increase, the followings are pointed out; ageing of the population, improved methods for detecting organisms from clinical specimens, increased physician's awareness on the disease, increased exposure of patients to the source of the organism. In Japan, it has been estimated that the overall incidence of NTM disease is about 3 per 100,000. About 80% of NTM are MAC, and among the remainder, Mycobacterium kansasii is most common in our country. Our hospital located in Fukuoka prefecture in Kyushu, western part of Japan. In this study, clinical data of 24 cases of pulmonary infection caused by Mycobacterium kansasii in our hospital, from 1996 to 2000 were investigated. Primary infection type patients were younger than secondary infection type. Nearly all secondary infection type patients had underlying diseases and complications. Serum total protein and albumin in primary type is lower than that in secondary type. The results of mycobacterial drug sensitivity tests were as follows; for rifampicin, 23 cases were sensitive to 10 micrograms/ml, all cases to 50 micrograms/ml, for ethambutol, 15 cases were sensitive to 2.5 micrograms/ml, 22 cases to 5 micrograms/ml, and for isoniazid, all cases were resistant to 0.1 microgram/ml, 11 cases were sensitive to 1 microgram/ml and 23 cases to 5 micrograms/ml. Sputum cultures of patients treated with drug regimens containing RFP converted to negative within 2 months after starting chemotherapy. Although three patients with serious complications died, other 21 patients improved and showed no relapse at least 6 months after the completion of treatment. PMID- 11855076 TI - A decade of successful tuberculosis control in New York City--the role of DOT vs DOTS. PMID- 11855077 TI - [How beds for tuberculosis be provided and utilized?]. AB - In 1951 when TB Control Law was legislated, and the government of Japan started intensive TB programme mainly consisting of mass health examination, BCG vaccination and distribution of appropriate treatment for TB cases, there were about 100,000 beds for TB, similar to the number of then TB deaths, and many TB patients died before admission to sanatoria. Urgent measures were taken to increase beds for TB with a target of 250,000, 2.5 times of then TB death. The target was achieved in 1957. Thereafter, the number of beds for TB as well as the occupancy rate had decreased with the decline of TB, and then policy on beds for TB could be summarized as follows: (1) top priority was given to increase the number of beds for TB, (2) general hospitals were improved with the progress of medical science and economic development, while no improvement was done on TB beds with the assumption that the need for TB beds will soon disappear, (3) minimum unit of TB beds was a TB ward with generally 40 to 50 beds, (4) an idea to provide TB bed in a general hospital came out only since 1992 as a small model project, (5) it was intended to segregate infectious TB patients from the community, however, no consideration was made about super-infection among patients themselves and the infection to health care workers, (6) admission of TB cases to a general bed and admission of non-TB cases to a TB ward was not legally permitted, (7) cost for TB treatment was set on a low level. Recent data indicate that the occupancy rate of TB beds was 43.5%, and the average stay in TB beds is still slightly over 100 days, and observing by prefectures, marked differences were seen. Taking into account changes in the pattern on TB patients such as aging and the increase of cases with serious complications and most health care workers in TB wards are not yet infected with TB, it is needed to divide TB beds into two types, one for new cases and the other for chronic cases. Beds for new cases should be provided in principle as a single room in a general hospital with good ventilation system, and DOT should be started in a hospital. Stay in this type of bed should not exceed 2 months, and higher medical fee should be provided. Beds for chronic cases could be provided in a TB ward. MDRTB cases are admitted in bed for chronic cases, however, preferably in a single room, and if active intervention such as chest surgery is tried in a few sophisticated hospital, medical fee for acute bed should be applied. Now, we have to change our mind from old concept of beds in TB ward to a TB bed in a single room with good ventilation. PMID- 11855078 TI - [Pulmonary complications in patients with AIDS]. AB - HIV infection was first reported in 1981 in USA. It has been 20 years since then. Owing to understandings of pathogenesis of this disease and development of new drugs such as the HIV-specific protease inhibitor (PI), prognosis of disease has been tremendously improved. Especially after 1997 in Japan, the strategy of anti HIV treatment shifted from two drugs combination to three drugs combination, which is called highly active antiretoviral therapy (HAART). HAART was so effective that prevalence of HIV associated opportunistic infections were decreased dramatically. Mortality among hospitalized HIV-infected patients was decreased from 6.7% in 1996 to 2.6% since then in ACC. However, 80% of patients receiving HAART suffered from side effects and 15% of them had to be changed their treatment due to side effects. Furthermore, an unexpected side effect, namely lipodystrophy syndrome (LDS), was emerged among patients who were receiving HAART more than one year. LDS was first reported as re-distribution of lipid such as central obesity with or without lipo-atrophy from extremities and/or face. Now only cosmetic change, but also it is associated with elevation of lipid and glucose level. Therefore, those patients who have LDS are in face of the risk for the ischemic heart diseases. Our survey indicated that the rate of LDS in Japanese patients were almost same as that of Caucasian patients reported elsewhere. Opportunistic infections associated with HIV infection Treatment for HIV infection consists of two major arms; one is use of anti-HIV drugs to prevent development of AIDS described above and the other is diagnosis, treatment, and prophylaxis of opportunistic infections. There are five very important opportunistic infections; Pneumocystis carinii pneumonia (PCP), cryptococcus meningitis, toxoplasma encephalitis, cytomegalovirus (CMV) infection, and Mycobacterium avium complex (MAC) bacteremia. Because if these five were able to diagnose, a patient can survive under appropriate treatment. On the other hand, if these were not diagnosed, patient must be AIDS death. After introducing HAART, number of CMV retinitis, MAC bacteremia, and AIDS dementia complex were decreasing. However, number of PCP sustained high because PCP is the first indicator disease of AIDS if the patient did not know his HIV status. The first choice of drug is sulfamethoxazole/trimethoprim (ST) for PCP treatment. If the patient were in severe respiratory failure, corticosteroid is used concomitantly. Treatment is usually continued for 3 weeks. We have successfully treated 45 out of 47 cases of PCP for 4 years. However, those patients treated with ST for 3 weeks were limited only 35% because of very high rate of side effects of ST. If the patient was intolerant to ST, treatment was switched to pentamidine. After finishing the treatment, the patient is to be treated with a 5-day course of oral desensitization to ST. More than 80% of patients who were previously intolerant to ST became successfully getting tolerance by this method. PMID- 11855079 TI - [Clinical significance of serum KL-6 in pulmonary sarcoidosis]. AB - We investigated whether the level of serum KL-6 could be an activity marker for pulmonary sarcoidosis. In 33 patients with pulmonary sarcoidosis, the relationships between serum KL-6 levels and diagnostic imaging, serum angiotensin converting enzyme (ACE) levels, serum lysozyme levels, steroid therapy, and prognosis were evaluated. There were no significant differences in the level of serum KL-6 when the patients were divided on the basis of radiographic findings, but the level of serum KL-6 was markedly elevated in some patients with stage-II pulmonary sarcoidosis. There was a significant correlation between serum KL-6 levels and the following two parameters: serum ACE and lysozyme levels. Among patients with a high initial level of serum KL-6, pulmonary sarcoidosis tended to become exacerbated within one year. Steroid therapy significantly decreased the level of serum KL-6, suggesting that the level of serum KL-6 could be an activity indicator for pulmonary sarcoidosis. Immunohistochemical staining by anti-KL-6 antibody revealed that KL-6 was localized in proliferating type-II alveolar epithelial cells. PMID- 11855080 TI - [Comparison of five guidelines of community-acquired pneumonia]. AB - The aim of this retrospective study was to elucidate the characteristics of five guidelines of community-acquired pneumonia: ATS (1993), ATS (2001), IDSA (1998), IDSA (2000) and the guidelines of the Japan Respiratory Society (2000). One hundred community-acquired pneumonia patients admitted to the International Medical Center of Japan were investigated in accordance with each set of guidelines based on the physical, laboratory, and chest radiography findings on the first day of treatment. According to the ATS (1993) guidelines, 33% of the cases were classified as "severe" pneumonia. On the other hand, according to the ATS (2001) guidelines, only 8% of the cases were classified as "severe" pneumonia. According to the IDSA guidelines, 35% of the patients were classified as "outpatients". Fluoroquinolone appears to be a very important antibiotic drug in the new guidelines of both ATS and IDSA. The scoring system of IDSA suggested a correlation between the patient's score and the pathogenic bacteria. According to the guidelines of the Japan Respiratory Society, 42% of the cases were classified as "severe" pneumonia. There are evident differences between these guidelines, and clinicians need to have a full understanding of their respective characteristics. PMID- 11855081 TI - [Clinical evaluation of 8 patients with pulmonary tuberculosis in a community hospital without restricted tuberculosis wards]. AB - The clinical features of 8 patients with pulmonary tuberculosis under treatment in the Koebaru Central Hospital, a community hospital without restricted tuberculosis wards, between 1992 and 2000 were evaluated. The major findings in the present study were: i) recently, the number of elderly patients with reactivated pulmonary tuberculosis has increased, ii) none of the patients showed respiratory symptoms, and 3 patients had negative Mantoux skin tests, iii) only one patient showed cavities on chest radiography, and iv) none of the expectorated sputum samples was smear-positive for Mycobacterium tuberculosis. Smears of intrabronchial sputum sampled using a fiberoptic bronchoscope from pulmonary lesions in patients without any symptoms were useful for establishing the diagnoses. Considering all aspects, it is important to perform aggressive evaluations, even for elderly patients, utilizing a fiberoptic bronchoscope for early diagnosis of pulmonary tuberculosis and to prevent nosocomial infections. PMID- 11855082 TI - [A case of inflammatory endotracheal polyps in an asthmatic subject]. AB - A 60-year-old asthmatic woman was admitted to our department because of bloody sputum and pneumonia. She had been treated with inhaled becromethasone dipropionate (800 micrograms/day) on an outpatient basis for 3 years. Fiberoptic bronchoscopy revealed polypoid lesions in the trachea, most of which were removed with forceps during the procedure. Numerous lymphocytes were observed in the biopsy specimen. Because immunohistochemical staining denied a monoclonal origin for the accumulated lymphocytes, the lesion was diagnosed as an inflammatory polyp. The patient was treated successfully with antibiotics for her pneumonia, and on a follow-up bronchoscopy 6 months later, only a small remnant of the lesion was noted. This is the fourth report about inflammatory polyps in asthmatics. In the previous 3 cases, however, marked eosinophil infiltration was consistently reported. The lymphocyte predominance in the present case therefore suggests a distinct etiology rather than asthmatic airway inflammation. PMID- 11855083 TI - [A case of ankylosing spondylitis associated with pulmonary aspergillosis and amyloidosis]. AB - A 56-year-old man was admitted because of diarrhea, cough, weight loss, and disturbance of consciousness. He had been diagnosed as having ankylosing spondylitis at 18-years old. The spondylitis progressed until there was complete rigidity of the spine including the neck, hip and knee joints. Human leukocyte antigen (HLA) B27, which has been characteristic of ankylosing spondylitis, was also present in this case. A chest radiograph showed pleural thickness and a cavity in the right upper lobe; and a soft tissue mass and fluid level was found in the cavity. Aspergillus fumigatus was detected in the sputum and pulmonary aspergillosis was diagnosed. Biopsy of the colon revealed that a large interstitial amyloidosis. Despite the treatment of the patient's malnutrition and lung aspergillosis using amphotericin B, the clinical course was rapidly progressive and the patient died of respiratory failure due to lung aspergillosis. It is important to be aware of these rare complications, which are correlated with the prognosis in cases of ankylosing spondylitis. PMID- 11855084 TI - [A case of pulmonary eosinophilic granuloma arising rapidly 30 years after the start of smoking and remitting spontaneously without smoking cessation]. AB - A case of pulmonary eosinophilic granuloma which arose rapidly after 30 years of smoking and remitted spontaneously without smoking cessation is reported. The patient was a 54-year-old man complaining of a dry cough who had been smoking 30 cigarettes a day for 30 years. Chest roentgenography showed multiple nodular shadows and cystic lesions in the upper and middle fields of both lungs. Chest computed tomography revealed multiple small cysts and small nodular lesions, mainly in both upper lung fields. CT findings strongly suggested pulmonary eosinophilic granuloma. A transbronchial lung biopsy (TBLB) was performed and 4 specimens were obtained, of which 3 showed granulomatous lesions with eosinophils and histiocytes. Furthermore, the granulomatous lesions were positive for S-100 protein staining. The symptoms and radiographic findings improved markedly within about 6 months after the onset of symptoms without treatment. Many cases of this disease were diagnosed in the past by open lung biopsy, but the number of cases diagnosed by TBLB is now increasing. The effectiveness of open lung biopsy has been emphasized in the diagnosis of pulmonary eosinophilic granuloma, but TBLB is also useful for diagnosis, especially in the active or early stage of the disease. PMID- 11855085 TI - [An adult case of mediastinal tuberculous lymphadenitis]. AB - A 40-year-old woman was admitted to the hospital with general fatigue and cough. Chest CT films revealed mediastinal lymphadenopathy with multiple low density areas, but no pulmonary lesions. There were no abnormal findings on neck, abdominal or pelvic CT. A PPD skin test was strongly positive, but M. tuberculosis bacilli were not found in the sputum. Mediastinal tuberculous lymphadenitis was diagnosed histologically and bacteriologically from specimens obtained by mediastinoscopy. Fiberoptic bronchoscopy did not reveal tracheobronchial tuberculosis. Follow-up chest CT three months after the start of antituberculosis chemotherapy with isoniazid, rifampicin and ethambutol hydrochloride showed that the mediastinal lymph nodes were decreased in size. Mediastinal tuberculous lymphadenitis in adults is rare, but the number of reports has increased. Mediastinal tuberculous lymphadenitis in adults must be distinguished from other causes of mediastinal masses. In this case, mediastinoscopy was very useful for differential diagnosis. PMID- 11855086 TI - [Pulmonary involvement, pleural effusion, and electrocardiographic abnormality in hypereosinophilic syndrome]. AB - A 47-year-old man was admitted to our hospital with complaints of cough and shortness of breath. Chest radiography showed infiltration of the right lung and left pleural effusion, the eosinophil count increased notably in the peripheral blood, sputum, and pleural effusion. Transbronchial lung biopsy revealed the invasion of eosinophils like eosinophilic pneumonia. Heart failure easily developed in this patient after the intravenous infusion. Myocardial involvement was suspected, and hypereosinophilic syndrome was diagnosed. After prednisolone was administered, the peripheral blood eosinophil count normalized rapidly, and subsequently, the pleural effusion and infiltration shadows in the lung disappeared. PMID- 11855087 TI - [A case of eosinophilic pneumonia possibly due to ifenprodil]. AB - A 69-year-old woman with a history of subarachnoid hemorrhage was started on ifenprodil for dizziness. Three weeks later, fever, cough, chills, dyspnea and skin eruption developed. A chest radiograph showed bilateral ground-glass shadows. Blood tests showed a white cell count of 14,400/mm3 with 32% eosinophils and a C reactive protein (CRP) level of 20 mg/dl. The arterial blood gases on room air were as follows: pH 7.45, PaCO2 33 torr, and PaO2 56 torr (Table 1). Ifenprodil was withdrawn and intravenous meropenem and minocycline administration was started on admission. Her fever improved rapidly and the CRP decreased, but hypoxemia and hypereosinophilia persisted. On the third hospital day, she underwent bronchoscopy with bronchoalveolar lavage (BAL). The differential count of BAL cells was 63% eosinophils, 15% lymphocytes, 21% macrophages, and 1% neutrophils. Intravenous methylprednisolone 250 mg/day for 3 days was commenced, leading to a clinical improvement. She received oral prednisolone (30 mg/day) for the next 4 days, and was then discharged without any symptoms. She has had no recurrence since. Both the drug lymphocyte stimulation test and the skin test for ifenprodil were negative. PMID- 11855088 TI - [Spinal cord sarcoidosis without abnormal shadows on chest radiography or chest CT diagnosed by transbronchial lung biopsy]. AB - The patient was a woman aged 56 years. In February 1998, she complained of fatigability of the right upper limb and disturbed extension of the right fourth finger. Because her condition deteriorated gradually and myelopathic signs such as difficulty in walking developed, she was hospitalized in May 1999 for close examination and appropriate treatment; and she was a few days later transferred to our hospital because of progressive myelopathy. In T2-weighted MR images of the cervical spine, the high-intensity area ranged between C2 and Th1, and in Gd DTPA enhanced MRI the high-intensity area was seen between C3 and C7. Although chest radiographs and chest CT scans were normal, spinal cord sarcoidosis could not be ruled out, and therefore, bronchoscopic examination was performed. Specimens obtained from transbronchial lung biopsy (TBLB) revealed non-caseating epithelioid cell granulomas, and so and the disease was diagnosed as spinal cord sarcoidosis. Both symptoms and MRI findings were improved by treatment with corticosteroids. It is suggested that, in patients suspected of spinal cord sarcoidosis from MRI findings. TBLB should be aggressively attempted, even if chest radiographs and chest CT scans are normal. PMID- 11855089 TI - [A case of multiple emphysematous bullae treated with living-donor lung transplantation from identical twin brothers]. AB - A 30-year-old man with multiple emphysematous bullae and bronchiectasis was admitted to Toyama Medical and Pharmaceutical University Hospital because of exertional dyspnea and fever. His chest radiograph and CT scan revealed multiple large bullae in the right lung and infiltrative shadows in the left middle lung field. Pseudomonas aeruginosa was isolated from his sputum culture. Although standard therapies including various antibiotics were administered, his respiratory condition was exacerbated, accompanied with the enlargement of bullae in the right lung and the consequent shift of the mediastinum to the left. The patient and his family proposed lung transplantation, and we concluded that lung transplantation would be an appropriate treatment for his disease. We transported the patient to Osaka University Hospital. Living-donor lung transplantation from the patient's identical twin brothers was successfully performed. PMID- 11855090 TI - [A case of bronchiolitis obliterans: auscultation leading to accurate diagnosis]. AB - This is a case of a 31-year-old man with a history of common cold. He had been suffering from productive cough and dyspnea on exertion. Squawks were heard through auscultation, and hyperinflation was also observed in his chest radiograph. Bronchiolitis was first suspected as a result of HRCT and TBLB. He was then treated with CAM for six months, but his symptoms showed little improvement. So we evaluated his squawks objectively by phonopneumograph, performed video assisted thoracoscopic surgery and diagnosed his illness as constrictive bronchiolitis. Based on the examinations, PSL therapy was applied to his case, which contributed improving his condition remarkably. The significance of this case is that proper auscultation led to the accurate diagnosis. PMID- 11855091 TI - [A case of congenital esophagobronchial fistula discovered incidentally on esophagography]. AB - A 50-year-old man was admitted to our hospital because of an abnormal shadow, detected during an upper gastrointestinal examination. A chest radiograph showed an infiltrating shadow in the right middle lung field. A chest CT showed a fistula communicating between a bronchus (rt. B6) and the middle of the esophagus. Resection of the fistula was performed by video-assisted thoracoscopic surgery (VATS). Isolation of the fistula was straightforward, and there was no evidence of inflammation or adherent lymph nodes around it. Histologic examination of the resected specimen revealed that the fistula lumen was covered with squamous epithelium and muscularis mucosa. These findings suggested that this case could be categorized as Braimbridge type II. In this case, the chest CT showed the esophagobronchial fistula clearly, and was useful for the diagnosis. PMID- 11855092 TI - [A case of renal cell carcinoma complicated with interstitial pneumonitis, complete A-V block and pleural effusion during interferon-alpha therapy]. AB - A 76-year-old man with postoperative renal cell carcinoma accompanied by multiple lung metastasis was being treated with recombinant interferon-alpha. After administration of 3 MU/day on 3 days/week for 1 month, he complained of headache and tinnitus. During continuous treatment for 3 months, he complained of appetite loss, low-grade fever and dyspnea. He was then referred to our Department of Internal Medicine. Electrocardiography indicated a complete A-V block, and chest radiography (CXR) showed a reticular shadow in both lower lung fields and bilateral pleural effusion. Chest computed tomography (CT) indicated subpleural emphysematous changes, multiple nodules, consolidation shadow with ground glass opacity in both lower lobes, and bilateral pleural effusion. The findings in the bronchoalveolar lavage (BAL) fluid included increases in the numbers of lymphocytes and eosinophils. We reached a diagnosis of interferon-alpha-induced pneumonitis on the basis of the patient's clinical course, and the CXR, chest CT and BAL fluid findings. Treatment with methylprednisolone pulse therapy for 3 days and then administration of prednisolone for 1 month resulted in marked improvement in the complete A-V block and interstitial pneumonitis. At day 7 after discontinuation of prednisolone, the serum level of C-reactive protein increased, and CXR showed bilateral pleural effusion. We therefore believe that the pleural effusion was probably also induced by interferon-alpha. Interferon is an effective drug for chronic hepatitis C and malignant diseases. Many complications have been reported during interferon therapy. However, although these complications, such as interstitial pneumonitis, complete A-V block and pleural effusion, have rarely been reported, careful attention is required during interferon therapy in case any appear. PMID- 11855093 TI - Double stent for malignant combined esophago-airway lesions. AB - OBJECTIVE: Combined esophago-airway stenosis and/or esophago-airway fistula due to malignancy bodes a dismal prognosis. We describe our work with double stents for combined esophago-airway lesions. METHODS: Between February 1994 and July 2000, we treated 11 patients using double stents--the Dumon stent for the airway and the covered Ultraflex for the esophagus. Double stenting was necessitated by combined esophago-airway stenosis in 8 patients and fistulas in 3, of these, 6 had lung cancer and 5 esophageal cancer. RESULTS: In all but 1 ventilator dependent patient, dyspnea and dysphagia were significantly reduced and fistula was successfully closed after double stenting. This palliation effectively continued more than 1 month in 5 patients, more than 2 months in 3, and more than 3 months in 2. Mean survival was 64 days (range: 9 to 148 days). Life-threatening complications developed in 5 (45%)--massive bleeding in 3 and uncontrollable esophago-airway fistula in 2. All 5 had received prior radiation. CONCLUSION: Although patients who received radiation frequently had life-threatening complications after double stenting, this procedure improved the quality of life in patients with esophago-airway stenosis or fistulas due to lung or esophageal cancer. PMID- 11855094 TI - Video-assisted lobectomy in elderly lung cancer patients. AB - OBJECTIVES: We evaluated the pre-, intra- and postoperative outcome of video assisted thoracic surgery lobectomy in elderly lung cancer patients to determine what factors may be disadvantageous. METHODS: From June 1982 to May 2000, 707 patients underwent pulmonary resection for primary lung cancer. Of these, 87 patients with t1-2 peripheral lung cancer underwent lobectomy and postoperative pulmonary function tests and postoperative conditions at an average of 2.3 months postoperatively. Of these, 52 underwent video-assisted thoracic surgery lobectomy since 1994 and 35 lobectomy by standard thoracotomy. RESULTS: Video-assisted thoracic surgery lobectomy offered advantages in blood loss, chest wall damage, and minimal performance deterioration status. The percent vital capacity, percent forced expiratory in 1 second, and percent maximum ventilatory volume were well preserved in patients who underwent video-assisted thoracic surgery lobectomy. Multivariate logistic regression analysis identified operation duration as an independent risk factor in morbidity and operative procedure as an independent risk factor in performance deterioration. In stage IA and IB patients, 3-year survival was 92.9% and 5-year survival 53.8% in those undergoing lobectomy by standard thoracotomy and 84.2% at 3 years and 60.1% at 5-years in those undergoing video-assisted thoracic surgery lobectomy. CONCLUSION: We thus consider video-assisted thoracic surgery lobectomy in this age group to be an effective procedure, but the long surgical duration is a risk factor in a poor clinical outcome. PMID- 11855095 TI - Cardioprotective effect of ischemic preconditioning on global myocardial ischemia in a sheep right heart bypass model. AB - OBJECTIVES: Ischemic preconditioning has been used to induce the myocardium to adapt to ischemic stress preceded by short periods of ischemia and reperfusion. We used a sheep right heart bypass model with a conductance catheter to assess the cardioprotective effect of ischemic preconditioning on 30-minute normothermic global myocardial ischemia. METHODS: Ischemic preconditioning was conducted in 6 sheep in 35-minute aortic cross-clampings interspersed with 5 minutes of reperfusion during cardiopulmonary bypass, with 6 sheep as time-matched controls. Global myocardial ischemia was subsequently achieved in 30-minute aortic cross clamping with left ventricular unloading during normothermic cardiopulmonary bypass. Weaning from cardiopulmonary bypass was conducted 40 minutes after reperfusion. Before ischemia and 40, 70, and 100 minutes after reperfusion, left ventricular pressure-volume loops were measured using a conductance catheter during right heart bypass preparation. Left ventricular contractility, diastolic function, and mechanical efficiency were then evaluated. Right heart bypass was instituted to control the preload and to decompress the right ventricle completely, thereby eliminating parallel conductance variation. RESULTS: No differences in the studied parameters were seen between ischemic-preconditioning and control groups before ischemia. Left ventricular contractility, diastolic function, and mechanical efficiency in the ischemic-preconditioning group were significantly superior to those in the control group after reperfusion. CONCLUSIONS: Ischemic preconditioning attenuates postischemic myocardial dysfunction in a sheep model using 30-minute unloaded normothermic global myocardial ischemia. Ischemic preconditioning would thus be clinically significant when the ischemic damage is severe. PMID- 11855097 TI - Left atrial free-floating ball thrombus with recent cardioembolic stroke. AB - A 67-year-old man was admitted to a local hospital complaining of hemiparesis. Because of coexisting arrhythmia, he was examined by echocardiography and found to have an oval free-floating thrombus in the left atrium concomitant with mitral and aortic stenosis. He was transferred to our hospital for emergency surgery. He underwent an operation on the ninth day from the onset of neurological dysfunction, when a 2.5 x 2.5 x 3.0 cm ball thrombus was removed, and the diseased mitral and aortic valves were replaced. His postoperative course was uneventful, with no neurological sequelae. Though left atrial ball thrombus is rarely found in patients with mitral valve disease, when it is found, then immediate surgical intervention is recommended to avoid sudden death. However, there is a high risk that any cerebral lesion may worsen due to systemic heparinization. Therefore, the optimal time of surgery in a patient with a recent neurological deficit is controversial. PMID- 11855096 TI - Endoscopic closure of bronchopleural fistula after pneumonectomy by submucosal injection of polidocanol. AB - We report two cases of a bronchopleural fistula with, and without, empyema treated by endoscopic submucosal injection of polidocanol (sclerotherapy) and application of cyanoacrylate. Case 1: A 60-year-old man underwent left pleuropneumonectomy for lung cancer. He developed bronchopleural fistula with empyema at 32 days after the operation. We performed sclerotherapy around the fistula. The air leakage stopped at 2 weeks after the sclerotherapy, and the fistula was closed. He was eventually cured of the empyema by pleural drainage. Case 2: A 61-year-old man underwent left pneumonectomy for lung cancer. He developed bronchopleural fistula without empyema at 50 days after the operation. We performed sclerotherapy and application of cyanoacrylate. After this therapy, the air leakage stopped immediately, and the bronchopleural fistula was closed. The sclerotherapy and application of cyanoacrylate are not only technically easy, but also very effective for treatment of bronchopleural fistula. Sclerotherapy and cyanoacrylate may be advocated as a first therapeutic step. PMID- 11855098 TI - Cystic tumor of the atrioventricular nodal region. AB - We report a case of a 66-year-old female who presented exertional dyspnea and palpitation. Echocardiography, transesophageal echocardiography and computed tomography showed a right atrial cystic mass attached to the interatrial septum. The patient underwent successful excision of the mass. The histopathological findings confirmed the lesion as a cystic tumor of the atrioventricular nodal region. This is the third known case of this condition diagnosed antemortem and treated successfully with surgical excision. PMID- 11855099 TI - Gigantic thymolipoma. AB - A 48-year-old woman with an abnormal shadow in chest radiography during an annual physical examination was found by chest computed tomography to have a large fatty mass lesion found to be diagnosed as a gigantic lipoma. Histopathological diagnosis was found to be benign thymolipoma consisting of mature fatty tissue and hyperplastic thymic tissue structures with Hassall,s corpuscles. Although the diagnosis is supported by imaging studies that demonstrate fat and soft tissue within the tumor, variations occur in computed tomography appearance. We suggest that surgical excision be considered when a gigantic intrathoracic lipomatous mass is in scanning as in this case. PMID- 11855100 TI - Aortic aneurysm and aortic regurgitation following aortic valve replacement due to Takayasu's arteritis. AB - A 26-year-old man who underwent aortic valve replacement for aortic regurgitation due to Takayasu's arteritis 2 years earlier experienced left amaurosis persisting for some minutes. Computed tomography showed aneurysmal dilation of the ascending aorta to a diameter of 60 mm and occlusion of the left carotid artery. Cardiac echography showed perivalvular leakage. Following administration of a calcium antagonist, the patient's amaurosis subsided and brain bloodstream scintigraphy showed no abnormalities. We resected the aneurysm instead of using Bentall's operation. Following an uncomplicated postoperative course, the patient was discharged 21 days after surgery and echocardiography has shown no perivalvular leakage to date. PMID- 11855101 TI - Subcarinal bronchogenic cyst with high carbohydrate antigen 19-9 production. AB - In a rare case of bronchogenic cyst with high carbohydrate antigen (CA) 19-9 production, a 57-year-old man with coughing and chest pain was diagnosed with a subcarinal mediastinal tumor. Fiberoptic bronchoscopy showed an erosive mucosal lesion overlying the area of extrinsic compression at the membranous of the right mainstem bronchus. Serum carbohydrate antigen 19-9 was elevated at 1300 U/ml. Thoracotomy showed an encapsulated cyst tightly adhering to the right main bronchus. The cyst was extirpated after ablation at the adherent cystic wall by electrocautery. Although intracystic carbohydrate antigen 19-9 concentration was very high, serum carbohydrate antigen 19-9 and bronchoscopic findings returned to normal postoperatively. The histological diagnosis was consistent with a bronchogenic cyst and carbohydrate antigen 19-9 could be immunohistochemically demonstrated within its epithelium. PMID- 11855103 TI - Cross-cultural nursing research: a two-way street. PMID- 11855102 TI - Ultrastructural damage to the preserved lung and its function after reperfusion. AB - OBJECTIVE: This study was undertaken to clarify what damage to a lung during cold storage influenced the function of transplanted lung after reperfusion. METHODS: We examined the ultrastructural damage in preserved right lung before reperfusion, and the function of transplanted left lung, in a same dog and measured the pulmonary artery oxygen pressure after reperfusion and the wet-to dry-weight ratio. We compared these findings between those dogs that survived until six hours after reperfusion (Alive Group) and those dogs that did not survive (Dead Group). We also investigated any correlation between the ultrastructural damage in the preserved lung and the function of the transplanted lung. RESULTS: The frequency of protrusion and destruction of the endothelial cells in the small pulmonary artery, and vacuolization of pneumocytes, in the Dead Group was significantly higher than that in the Alive Group. A correlation was found between the frequency of two kinds of ultrastructural damage; vacuolization in the endothelial cells in the small pulmonary artery and vacuolization in the pneumocytes, and the pulmonary artery oxygen pressure at 1 hour after reperfusion. A correlation was also found between the frequency of the vacuolization of pneumocytes and the wet-to-dry-weight ratio. CONCLUSIONS: Findings suggested that a lung suffering severe damage to intracellular structure during hypothermic preservation is unable to function sufficiently after reperfusion and is at high risk for early graft failure. PMID- 11855104 TI - Accompany the sick: a unique practice in Chinese hospitals by patients' relatives and friends. AB - There is a practice commonly found in many Chinese hospitals called 'accompany the sick' (pei ban). It means that many patients' relatives or friends 'live' with patients in the hospital. Such practice can benefit patients in that the visitors are providing psychological support and carrying out some bedside nursing care to the patients. However, it also creates many problems to nurses and hospitals such as overcrowding, an over-demand of hospital resources, and maintaining quality care. There is no study of such practice. In order to gain better understanding of 'accompany the sick', its impact on nursing practice and nursing professional development, identifying the contributing factors of such practice is needed. This is an ethnographic case study of a public hospital in western China. The study has discovered a number of factors that contribute to the practice, such as the Chinese concept of duty of care, the geographic and economic situation, and the unique culture of nursing practice. This is a pilot study and the result of this study will contribute to further study in nurses' work in China. PMID- 11855105 TI - The change from general nurse education to a nursing diploma in Papua New Guinea. AB - Papua New Guinea has started to change from a hospital based general nurse training to a competency based, university linked, diploma of nursing training program. This paper looks at the development of nurse education in Papua New Guinea. It also briefly outlines the present and proposed courses. The expected benefits and anticipated problems are then discussed and the paper ends with challenges being faced by those involved in nurse education. PMID- 11855106 TI - The diabetes health care of Aboriginal people in South Australia (Part 1). AB - Although type 2 diabetes is a recognised health priority in South Australia, Aboriginal people with diabetes do not utilise the mainstream diabetes health services on a regular basis for health care. This means that Aboriginal clients have the potential to develop diabetes-related problems and, furthermore, are not in a position to make informed decisions about health care issues. This lack of client empowerment is contrary to the goals of contemporary diabetes health care and, as a result, Aboriginal clients suffer the consequences of ineffective management with a compromised lifestyle. To identify how this situation might be improved, a qualitative study funded by Diabetes Australia was undertaken in South Australia. The overall goal was, firstly, to identify the reasons why Aboriginal people with diabetes do not attend mainstream health agencies on a regular basis and secondly, if possible, to improve attendance. Thus, Aboriginal health professionals (n = 43) were recruited from the 8 statistical divisions of South Australia and interviewed about Aboriginal diabetes health care issues. In Part 1 of this series, the research findings indicated the beliefs and attitudes held by clients about diabetes, their lack of knowledge about management issues, their responses to diabetes, the effects of diabetes on their lifestyle and the strategies that diabetes health professionals used to help their clients deal with diabetes health issues. In Part 2 the research findings indicated the importance of the Aboriginal health worker to the successful diabetes management of Aboriginal clients, the constraints that affect the delivery of diabetes health care and the recommendations made by health professionals to improve the standard of diabetes health services. PMID- 11855107 TI - The diabetes health care of Aboriginal people in South Australia (Part 2). AB - Although type 2 diabetes is a recognised health priority in South Australia, Aboriginal people with diabetes do not utilise the mainstream diabetes health services on a regular basis for health care. This means that Aboriginal clients have the potential to develop diabetes-related problems and, furthermore, are not in a position to make informed decisions about health care issues. This lack of client empowerment is contrary to the goals of contemporary diabetes health care and, as a result, Aboriginal clients suffer the consequences of ineffective management with a compromised lifestyle. To identify how this situation might be improved, a qualitative study funded by Diabetes Australia was undertaken in South Australia. The overall goal was, firstly, to identify the reasons why Aboriginal people with diabetes do not attend mainstream health agencies on a regular basis and secondly, if possible, to improve attendance. Thus, Aboriginal health professionals (n = 43) were recruited from the 8 statistical divisions of South Australia and interviewed about Aboriginal diabetes health care issues. In Part 1 of this series, the research findings indicated the beliefs and attitudes held by clients about diabetes, their lack of knowledge about management issues, their responses to diabetes, the effects of diabetes on their lifestyle and the strategies that diabetes health professionals used to help their clients deal with diabetes health issues. In Part 2 the research findings indicated the importance of the Aboriginal health worker to the successful diabetes management of Aboriginal clients, the constraints that affect the delivery of diabetes health care and the recommendations made by health professionals to improve the standard of diabetes health services. PMID- 11855108 TI - Vision screening of school children: should it be continued? AB - Vision screening of all children at school entry has been a traditional practice for many years. Recently, decisions have been made to discontinue screening and to rely on parent and teacher referral methods instead. A review of the literature suggested that parent and teacher referral methods of screening were less than satisfactory, and that professional screening of all children at school entry age should be continued. PMID- 11855109 TI - The dis-ease of nursing academia: putting the vulnerability 'out there' (Part 1). AB - This article is the first in a two-part series on a recent research study investigating the lived experiences of women nurse academics in Australia. This article begins with a discussion on the growing necessity of nurses to care for one another while working in stressful environments. The author then discusses the need to research nurse academics. The majority of the article focuses on the methodological considerations associated with planning and researching the above topic. A comprehensive discussion of research methodology, methods and ethical concerns ensues. PMID- 11855110 TI - Is a PhD the best career choice? Nursing employers' views. AB - Recent debate has centred around a perceived discrepancy between employment skills required and skills acquired of doctorally prepared employees. There is a perception there might be a substantial shortfall between both current and future skill requirements. While much research has been undertaken into employer requirements of first degree graduates, there does not appear to have been any systematic survey/s of employer attitudes to research postgraduates (West, 1998). Ascertaining the views of those in the field likely to employ doctoral graduates is essential to ensure that research training is adapted to suit the workplace. This paper reports on a project that sought to elicit the opinions of some nursing employers about the suitability of doctorally prepared nursing employees. Results suggest that there might be some discrepancy between skills acquired at university and skills required in the workplace. While the study was small and generalisability is not claimed, we believe the results point to the need for further investigation. PMID- 11855111 TI - Nursing in a time of change: a case for critical thinking. AB - Like all professionals, nurses live with the inevitability of change. The extent to which change is experienced as a crisis rather than as an opportunity is determined by the degree of control that nurses have over the change process. This paper outlines the determinants of change, examines the impact of change on nursing practice, explores professional ways of knowing and proposes critical thinking as a means by which nurses can interpret and manipulate change and illustrates a curriculum approach that facilitates the acquisition of the capacity to think critically. PMID- 11855112 TI - Nursing as a profession. PMID- 11855113 TI - Mirror, mirror on the wall: who is the true nurse after all? AB - This somewhat colloquial reflection on personal experience reveals some commonly perceived images and metaphors pertaining to nursing that have been birthed in history, perpetuated by the media, and entrenched in the minds of both onlookers and, unfortunately, of those not quite so detached. The reader might identify with some, and be amazed at others, or be provoked on a similar journey of reminiscence and reconsideration of the images that have shaped and informed their practice. The literature presents the not always positive portrayal of nursing, and the resultant adverse effect this exerts not only upon the profession, but upon its practitioners as well. Thus is set forth the challenge of how to respond. PMID- 11855114 TI - Informed consent versus uninformed consent: evidence of good practice? PMID- 11855115 TI - Reflective practice: a critical incident. AB - Reflection identifies one's professional capabilities, increases knowledge of self and improves understanding and management of patients. Conducting reflective practice is conducive to professional development and growth. A complex medical and nursing management of a psychiatric emergency is illustrated through this critical incident. Psychiatric nurses play a key role in deescalating the aggressive behaviour of psychotic patients. Through reflection, the basic tools of psychiatric nurses are reviewed and the prime components of psychiatric therapy are discussed. PMID- 11855116 TI - A look behind the screens in Papua New Guinea. AB - Nursing care is affected by the culture we live in and even when nursing care is given the same way in different societies, it can be given that way for different reasons. This was found when doing a comparison between how certain aspects of nursing care were performed and viewed in Australia and Papua New Guinea. The framework for comparison was derived from part of Lawler's book (1991) Behind the Screens: Nursing, Somology, and the Problem of the Body. The feelings and accounts of staff and students working in Papua New Guinea (in 1997) are compared with those expressed by Australian nurses interviewed by Lawler in the mid 1980s. Interviews and observations in Papua New Guinea are summarised and then analysed looking at similarities and differences between the two cultures especially in giving body care and care after death. PMID- 11855117 TI - Clinical challenges that face nurses working in a correctional setting. AB - The number of people that have been incarcerated into the Australian prison system is increasing. Many of these people come to the system with complicated health needs. Nursing staff provide the first point of contact for their health needs, and it is the responsibility of the nursing staff to be made aware of inmates' needs and how to address these needs. These problems can present an enormous challenge to nurses working in a corrective setting. This essay has been written to demonstrate how nurses at this particular Corrective Centre overcome the problems they encounter in their everyday work environment. PMID- 11855118 TI - A case in point? A parasuicide patient's recollections of being nursed: a discourse analysis. AB - This paper highlights the recollections of being nursed from the perspective of a patient (Gabby) admitted to a general medical ward after attempting suicide. A discourse analysis was used to discuss the way in which Gabby was nursed, enabling identification of the social structure which supports the nurse-patient relationship. This research evolved with the realisation that the point of view of parasuicide patients was primarily absent within nursing literature. Inattention within the literature was later replicated by Gabby's expressions of inadequate interaction between nurses and parasuicide patients. PMID- 11855119 TI - The role of nursing unit managers in educating nurses. AB - This paper presents the findings of a work sampling study undertaken in New South Wales (Australia) to ascertain the amount of time spent by nursing unit managers directly educating nurses. The results indicate that they spend very little time in teaching, traditionally one of their role tasks. It is argued that their teaching at unit level should focus on their role as leader, mentor, preceptor and coach. This role shift needs to be articulated to managers and nurses to minimise confusion over the role and to ensure that direct teaching is placed into the hands of expert clinicians. PMID- 11855120 TI - Human resource management strategies for the retention of nurses. AB - Retention of nurses is a critical problem facing health care managers across Australia today. Retention of experienced employees is not only cost effective, but of great importance to the continued high performance of the nursing services in health care organisations. This article will examine innovative human resource management strategies to promote the retention of nurses. PMID- 11855121 TI - What you ask is what you get: learning from interviewing in qualitative research. AB - This paper outlines some important issues discovered about undertaking qualitative interviews. A description of a study that involved interviewing the authors' students is followed by discussion of aspects of the interviewing process that became apparent during analysis of those interviews. Problems that were encountered with retrospective interviews, power differentials, question sequencing, some interview techniques, and a prior knowledge of the students are illuminated to assist other qualitative interviewers to be aware of these potential areas of difficulty. PMID- 11855122 TI - Epistemology at work: the ontological relationship between feminist methods, intersubjectivity and nursing research--a research exemplar. AB - This article explores the importance of strong epistemological and ontological links in nursing research by examining the design and process of a recent research project. The research topic concerns the relationship between self concept and nursing practice. In this article, the authors demonstrate that commitment to a methodologically consistent process and the necessary associated epistemological and ontological positions provides a depth and structure to nursing research. It is the authors' belief that such consistency within research acts to strengthen the research process, and consequently strengthens nursing's research base and knowledge. PMID- 11855123 TI - Social impact of same day surgery for the patient and the carer: a study of patients' experiences. AB - A semi-structured questionnaire was used to interview 18 patients, prior to surgery, on the day of their admission to day surgery. The purpose of the interviews was to gather information about the social arrangements, for both patient and carer, that were made necessary by the need for the patient to attend surgery and to be cared for at home over the 24-hour post-discharge period. The initial response of the patients concerning the arrangements made for their post operative care didn't expose any far-reaching difficulties. However, their descriptive stories reveal the extent of the measures that some needed to take in order to attend surgery and be cared for at home. These results point to a need for extensive research into the cost of the social impact of day surgery on the patient and the carer. PMID- 11855124 TI - [Metabolic populational design of tumors in vivo in genetically susceptible individuals]. PMID- 11855125 TI - [Increase in the level of lipid peroxidation products as a risk factor for death in a prospective study]. PMID- 11855126 TI - [Interaction of the cerebral hemispheres in memorizing the rhythm of motion]. PMID- 11855127 TI - [Refusal phenomenon in muscle activity. The role of the respiratory system]. PMID- 11855128 TI - [Variability in heart rhythm: approaches to mechanisms]. PMID- 11855129 TI - [Effect of caffeine on cognitive function and psychophysiological status in man]. PMID- 11855130 TI - [Improvement of the method of monitoring cardiac activity during physical exercise]. PMID- 11855131 TI - [Use of adaptive biocontrol using EEG for correcting the functional status of neurology patients]. PMID- 11855132 TI - [The so-called alternative vision or the direct vision phenomon]. PMID- 11855133 TI - [Change in adrenal glucocorticoid function in first grade boys during adaptation to the beginning of learning in school and during the course of the school year]. PMID- 11855134 TI - [Features of EEG in women with complicated pregnancies]. PMID- 11855135 TI - [Interaction of the frontal and left parietotemporal cortex in verbal thinking]. PMID- 11855136 TI - [Role of functional asymmetry of the brain in forming a subjective assessment of status during hypoxic interval training]. PMID- 11855138 TI - [Monitoring function of the right heart ventricle during correction of an interventricular septal defect]. PMID- 11855137 TI - [Electrophysiological and vegetative indicators of human emotional response to dynamic color music]. PMID- 11855139 TI - [Internal rhythm of the electrocardiosignal and effectiveness of adaptation to cyclical muscular work]. PMID- 11855140 TI - [Some regularities in blood circulation in highly-skilled swimmers]. PMID- 11855141 TI - [Diagnosis of aging. II. Age dynamics of the correlation between the biological markers of aging]. PMID- 11855142 TI - [Experimental study of interleukin-12 gene vaccines in the treatment of low-load malignant lymphoma (EL4)]. AB - OBJECTIVE: Two kinds of murine interleukin-12 (mIL-12) fusion gene vaccines were used to treat the murine low-load malignant T cell lymphoma EL4 as minimal residual disease (MRD) model. METHODS: C57BL/6 synergistical mice were subcutaneously inoculated with 1 x 10(6) wild-type (wt) EL4 tumor cells as low load lymphoma model treated with two mIL-12 gene vaccines. Package cell line PA317/12 producing mIL-12 retrovirus (RV) was used as in vivo vaccine and EL4 tumor cells transferred with mIL-12 gene as ex vivo vaccine. RESULTS: In both mIL 12 gene vaccine-treated groups, there was no tumor growth in 50% mice 60 days after inoculation. Nine of these no tumor growth mice were re-challenged with 5 x 10(5) wt EL4 cells, and 5 of them survived without tumors in another 60 days. All control mice died with tumors within one month after inoculation. Among those developed tumors in both vaccine-treated groups, the development of tumors was delayed, the survival period prolonged (P < 0.01), and the tumors size at death smaller (P < 0.05) as compared with the controls. In the long-survived vaccine treated mice, no residual tumor cells were found by morphological examination. CONCLUSION: Both IL-12 gene vaccines can efficiently eliminate wt EL4 MRD in C57BL/6 mice. PMID- 11855143 TI - [Clinical and cytogenetic analysis of patients with lymphoid malignancies carrying 14q32 abnormality]. AB - OBJECTIVE: To investigate the clinical and cytogenetic characteristics of patients with lymphoid malignancies (LM) harboring 14q32 abnormalities. METHODS: The cases with 14q32 aberrations in 225 patients with various LM were analyzed by clinical data. RESULTS: Fifteen (6.67%) patients with abnormalities involving 14q32 were found and the pattern of 14q32 aberration was associated with different types of LM. t(8;14)(q24;q32) was the commonest karyotypic aberration and predominant in acute leukemias. There was no consistent profile morphologically and immunophenotypically in the involving leukemias, however there were common clinical and prognostic features and often with an additional characteristic abnormality of ins(1;6)(q11;q23q27). One case of myelodysplasia (MDS) secondary to multiple myeloma displayed t(7;14)(q34;q32) in primary clone and additional 7q- and 20q- in derivative clone that was frequently involved in MDS. CONCLUSIONS: 14q32 abnormalities and additional chromosomal alterations can facilitate the diagnosis and prognostic assessment for the involving LMs. PMID- 11855144 TI - [Study on structure change and hypermethylation of p16 gene in multiple myeloma]. AB - OBJECTIVE: To illustrate the role of structure and hypermethylation of p16 gene in the pathogenesis of multiple myeloma(MM). METHODS: By using PCR-single strand conformation polymorphisms(PCR-SSCP) and methylation-specific PCR(MSP) techniques, the structure and hypermethylation status of p16 gene in MM cell lines and patients were analysed. RESULTS: Homozygous deletion of p16 exon 2 was found in KM3 cells. The completely methylated p16 gene and hypermethylation of CPG island were observed in U266, LP1 cell lines and 55.56% of MM patients. CONCLUSION: Methylation of p16 gene is important in the pathogenesis of MM and may provide a new drug target for the treatment of MM. PMID- 11855145 TI - [The clonal origin of the myeloma cells secreting three kinds of immunoglobulin from a multiple myeloma patient]. AB - OBJECTIVE: To explore the clonal origin of myeloma cells from a multiple myeloma (MM) patient with 3 kinds of immunoglobulin (Ig). METHODS: Six primers for IgH variable region gene families VH1-VH6 and one primer of common join region (J) were used to perform reverse transcriptase polymerase chain reaction(RT-PCR). Five oligonucleotides of constant region gene (C mu, C delta, C gamma, C alpha and C epsilon) were also used as reverse primers. Different combination of primers were used to perform PCR. PCR products of IgH gene rearrangement were cloned and sequenced. RESULTS: A sharp and dense gene band appeared by using Ig VH3 gene family primer and J primer. The specific IgH gene bands also appeared by using primers of VH3 with C mu and C gamma and C alpha but not by primers of VH1, VH2, VH4, VH5 with J as well as VH3 with C delta and C epsilon. It indicated that there were IgM, IgG and IgA gene rearrangement. These gene products were cloned and sequenced. All 3 IgM, IgG and IgA gene sequences are identical. CONCLUSION: Despite the oligoclonality of MM cell, the sequencing results of 3 IgH genes showed that all the malignant cells were monoclonal origin. PMID- 11855146 TI - [The effectiveness of cyclosporin A in the treatment of autoimmune hemolytic anemia and Evans syndrome]. AB - OBJECTIVE: To test the therapeutic effectiveness of cyclosporin A (CsA) in combination with corticosteroid in the treatment of autoimmune hemolytic anemia(AIHA) and Evans syndrome. METHODS: Forty-four cases of AIHA and Evans syndrome were treated with CsA in combination with a conventional regimen (prednisone + danazol) (18 cases) or conventional regimen alone(26 cases). RESULTS: The complete response rate of CsA group (88.9%) was higher than that of conventional regimen group (57.7%) (P < 0.05), and the relapse rate was lower (3.3% vs 70%) (P < 0.01). CONCLUSION: CsA in combination with conventional regimen can improve the complete response rate and reduce relapse of AIHA and Evans syndrome. PMID- 11855147 TI - [In vitro effect of lovastatin on NB4 promyelocytic leukemia cells]. AB - OBJECTIVE: To explore the effect of lovastatin(LOV) on cell growth, apoptosis and differentiation of NB4 cells and elucidate some of the possible mechanism. METHODS: Cell proliferation and viability were analyzed by MTT assay and trypan blue exclusion assay, cell differentiation by NBT reduction. Cell morphological examination, DNA electrophoresis, flow cytometry, TUNEL in situ hybridization and semi-quantitative RT-PCR were performed to determine cell apoptosis. H,K,N-ras gene expression was investigated by RT-PCR and p21Ras protein expression by flow cytometry. RESULTS: 1. LOV inhibited the proliferation of NB4 cells with a lC50 of 12.59 mumol/L. 2. LOV induced apoptosis of NB4 cells, affected the cell cycle progression and arrested NB4 cells in G1/S phase. Bcl-2 mRNA was down-regulated by LOV in a time-dependent manner. 3. LOV could not induce the differentiation of NB4. 4. LOV did not increase the expression of H,K,N-ras mRNA, but decrease p21Ras protein expression on the cellular membrane. CONCLUSION: LOV can inhibit proliferation, induce apoptosis and interfere with cell cycle progression, but not induce differentiation of NB4. Inhibition of p21Ras protein isoprenylation is considered to be the major mechanism of LOV affecting NB4 cells. PMID- 11855148 TI - [Effect of interferon-alpha on the prognosis of bone marrow transplantation in chronic myeloid leukemia]. AB - OBJECTIVE: To investigate whether prior interferon alfa (IFN-alpha) treatment affects the outcome of allogeneic bone marrow transplantation. METHODS: The outcome of 85 patients with chronic myelogenous leukemia (CML) in first chronic phase received transplants from an HLA-identical sibling donor was analyzed. Of the 85 patients, 30 did not receive IFN-alpha, whereas 30, 15 and 10 patients received IFN-alpha 3 x 10(6) units 3-4 times per week for < 6, 6-12 and > 12 months before transplant combined with hydroxyurea and/or HA regimen (H: Harringtonine; A: Ara-C) respectively. RESULTS: Pretransplant IFN-alpha therapy for > or = 6 months was associated with an increased risk of severe (grade III IV) acute graft-versus-host disease (GVHD) (P < 0.01) as compared to that in < 6 months or no IFN-alpha therapy. No influence of pretransplant IFN-alpha treatment on engraft, cGVHD, VOD, relapse and TRM (transplant-related mortality) was found. Survival rate of IFN-alpha therapy < 6 months group (90.0%) was higher than that of no IFN-alpha group (68.9%) or > 12 months group(60.0%) (P < 0.05). CONCLUSION: It was found that there was a trend towards an improved survival in the IFN-alpha < 6 months group. Pretransplant IFN-alpha therapy for > or = 6 months was associated with an increased risk of severe aGVHD. PMID- 11855149 TI - [Analysis of telomerase activity and telomere length in acute myelogenous leukemia]. AB - OBJECTIVE: To study the changes and significance of telomerase activity and telomere length in acute myelogenous leukemia. METHODS: TRAP-ELISA-PAGE was used to detect telomerase activity, Southern blot to estimate the length of telomere. RESULTS: Telomerase activity was significantly higher in AML(Absorption(A): 2.298 +/- 1.059) than in normal control(A: 0.387 +/- 0.598) and the mean telomere length of AML [(7.6 +/- 2.1) kb] was significantly shorter than that of normal control[(9.3 +/- 1.9) kb]. The alteration of the telomere length was detected mainly in telomerase-positive AML. CONCLUSIONS: It suggested that there was a close relationship between telomerase activity and telomere length in AML. The activation of telomerase might play an important role in the genesis and development of AML and telomere length changes may correlate with the activation of telomerase. PMID- 11855150 TI - The effect of parental imprinting on the INS-IGF2 locus of Korean type I diabetic patients. AB - BACKGROUND: Insulin-dependent diabetes mellitus (IDDM) is caused by the autoimmune destruction of pancreatic beta-cells. Susceptibility to IDDM appears to depend on more than one genetic locus. Evidence of a genetic linkage for IDDM2 was found in male meioses from French and North American populations. It is linked to maternal imprinting (i.e. monoalleleic expression of the insulin gene) that is considered the most likely cause of these gender-related differences. IGF2 is expressed only in the paternal allele and, therefore, is considered a candidate gene for IDDM2 transmission because of its important autocrine/paracrine effects on the thymus, lymphocytes and pancreas. Nevertheless, it remains controversial whether the parental origin of IDDM2 influences IDDM susceptibility. METHODS: Using PCR and semi-quantitative RT-PCR, we analyzed the INS/Pstl + 1127 and IGF2/Apal polymorphisms and RNA expression level between Pstl (+/-) and Pstl (+/+) to determine genotype and allele-specific expression of the INS and IGF2 genes. RESULTS: INS/Pstl (+/+) and IGF2/Apal (+/-) were observed in 36 (97.3%) of 37 IDDM patients and in 29 (72.5%) of 40 IDDM patients, respectively. The presence of both IGF2 alleles in RNA was observed in 21 (91.6%) of 24 IDDM patients. Our results show a 3-fold increase in RNA expression from Pstl (+/-) allele over Pstl (+/+) allele. CONCLUSION: Our conclusion does not entirely exclude IGF2 as the gene involved in IDDM2, even though the parental effect of IDDM2 transmission is not related to IGF2 maternal imprinting. The INS genotype appeared mostly in the Pstl (+/+) homozygote and, therefore, we could not explain the INS imprinting pattern in Korean type 1 diabetic patients. Genetic differences between populations may account for the discrepancy between Korean type I diabetic patients and American or French type I diabetic patients. PMID- 11855151 TI - Expression of mucosal cyto-chemokine mRNAs in patients with Helicobacter pylori infection. AB - BACKGROUND: Helicobacter pylori-induced destruction of the gastroduodenal mucosal barrier is initiated with mucosal infiltration of inflammatory cells. Cytokines and chemokines have been suggested to play important roles in the migration and activation of these inflammatory cells into the mucosa. The present study aimed to investigate expression rates of cyto-chemokine mRNAs using gastric mucosal biopsy specimens. METHODS: In 98 patients infected with Helicobacter pylori, mucosal mRNA expression rates of cytokines (IL-1 beta, IL-6, and IL-10), C-C chemokines (macrophage inflammatory protein 1 alpha (MIP-1 alpha), and macrophage inflammatory protein 1 beta (MIP-1 beta), monocyte chemotactic and activating factor (MCAF), regulated on activation, normal T cell expressed and presumably secreted (RANTES)) and C-X-C chemokines (IL-8 and growth regulated a (GRO-alpha)) were examined using reverse transcription polymerase chain reaction (RT-PCR). RESULTS: The expression rates of mRNA for IL-8, GRO-alpha, MIP-1 alpha and RANTES were significantly more increased in H. pylori-positive patients than in H. pylori-negative patients. However, the expressions of IL-1 beta, IL-6 and IL-10 mRNA were statistically not different between two groups. After eradication of H. pylori, expressions of mRNA for three cytokines (IL-1 beta, IL-6 and IL-10), four C-C chemokines (MIP-1 alpha, MIP-1 beta, MCAF and RANTES) and two C-X-C chemokines (IL-8 and GRO-alpha) were significantly decreased. CONCLUSION: These results suggest that C-X-C chemokines and some C-C chemokines play important roles in H. pylori-associated peptic ulcer diseases. PMID- 11855152 TI - Predictors for benign solitary pulmonary nodule in tuberculosis-endemic area. AB - BACKGROUND: Solitary pulmonary nodule (SPN) may show different presentation in tuberculosis (TB)-endemic countries. The aim of this study was to identify clinical and radiological predictors favoring benign or malignant SPN in TB endemic region. METHODS: Two hundred one SPNs in 201 consecutive Korean patients were included (< 3 cm in diameter, all confirmed by pathology or bacteriology, 93 benign and 108 malignant diseases). For clinical parameters, age, sex, smoking status and amount, and past history of pulmonary tuberculosis and diabetes mellitus were investigated retrospectively. For radiological parameters, size, location, margin characteristics, presence of calcification, pleural tag, surrounding satellite nodule, cavitation, internal low attenuation, open bronchus sign, surrounding ground-glass opacity, enhancement pattern of the SPNs and mediastinal lymph node (LN) enlargement were analyzed on chest CT scans. RESULTS: Patients with a older age (60.7 +/- 9.6 vs 56.2 +/- 13.1, p = 0.008) and more than 40-pack years smoking (27.8% vs 14.0%, p = 0.017) were more frequently related with malignant than benign SPN. On chest CT scans, spiculated margin, contrast enhancement more than 20 Hounsfield unit and presence of pleural tag and mediastinal LN enlargement were more frequently observed in malignant than benign SPNs. In contrast to previous studies, satellite lesions (21.5% vs 1.9%, p < 0.001) and cavitation (20.4% vs 5.6%, p = 0.001) were more frequently seen in benign than malignant SPN. Positive predictive values of benignity were 90.9% and 76.0%, respectively, when satellite lesions and cavitation were found in cases of SPN. CONCLUSION: Satellite lesions and cavitation on chest CT scan could be useful predictors for benign SPN in TB-endemic areas. PMID- 11855153 TI - Percutaneous catheterization of the internal jugular vein for hemodialysis. AB - OBJECTIVES: The present study was aimed at evaluating the clinical experiences in the internal jugular venous catheterization for hemodialysis. METHODS: We retrospectively analyzed the data on internal jugular venous catheterization at Chonnam National University Hospital from May 2000 to February 2001. RESULTS: There were 132 uremic patients with a total of 150 attempts of internal jugular cannulation. Overall success rate was 90.9% with average puncture trials of 2.3 +/- 2.1. 124 (82.7%) of the catheterization attempts were made on the right side and 26 (17.3%) were made on the left. The catheters were left in place from 2 to 87 days with an average of 19.5 +/- 15.3 days per catheter. The dialysis sessions per catheter were from 2 to 58 with an average of 11.3 +/- 6.8. The mean blood flow during hemodialysis immediately after catheterization was 213.4 +/- 42.2 ml/min. Thirty two (21.3%) patients had early complications. These included carotid artery puncture (11.3%), local bleeding (4.7%), local pain (3.3%), neck hematoma (0.7%) and malposition of the catheter (1.3%). Seventeen (11.3%) patients had late complications. These included fever or infection (11.3%), inadequate blood flow rate (3.3%) and inadvertent withdrawal (2.0%). There was no catheter-related mortality. CONCLUSIONS: Our experiences revealed that the internal jugular vein catheterization is relatively safe and efficient for temporary vascular access for hemodialysis. PMID- 11855154 TI - Dyspnea and palpitation during pregnancy. AB - OBJECTIVES: Dyspnea and palpitation are common features of pregnancy. While several theories have been put forward to explain the etiology of gestational dyspnea and palpitation, there have been few systemic studies of its incidence, severity and time-course in a group of normal women. METHODS: We interviewed postpartum women, within 3 days after delivery, about dyspnea and palpitation. Separately from this interview, we performed 24-hour ECG monitoring for obstetric patients with palpitation before delivery. RESULTS: The subjects interviewed were 261 women, of whom 37.5 percent and 11.5 percent experienced dyspnea and palpitation, respectively. These symptoms had tendency to increase to term. The presence of arrhythmias could be documented in only 22% of patients having 24 hour Holter monitoring. CONCLUSION: Dyspnea and palpitation were common among normal pregnant women and had a tendency to increase to term. PMID- 11855155 TI - A single center's 30 years' experience of esophageal adenocarcinoma. AB - BACKGROUND: Adenocarcinoma of the esophagus has been reported to be increasing in incidence in a number of regions throughout the world, while the incidence of squamous cell carcinoma (SCCA) of the esophagus is mostly stable or decreasing. To evaluate the increasing tendency of adenocarcinoma of the esophagus. METHODS: We studied retrospectively the records of patients with histologically proven esophageal cancer between 1970 and 1999 at the Yonsei Medical Center. RESULTS: Total cases of esophageal cancer were 969 patients of which the cases of adenocarcinoma and SCCA were 27 patients and 918 patients, respectively. The ratio of esophageal adenocarcinoma to SCCA was 0.0375 in the 1970s, 0.0241 in the 1980s and 0.0292 in the 1990s. There was no statistical difference (p = 0.811) in the ratios of adenocarcinoma of the esophagus between the three consecutive 10 year groups. CONCLUSION: In conclusion, unlike the US and other western countries, it seems that the ratio of esophageal adenocarcinoma compared to SCCA has not increased among patients with esophageal carcinoma at the Yonsei Medical Center. PMID- 11855156 TI - Is cholecystectomy necessary after ERCP for bile duct stones in patients with gallbladder in situ? AB - BACKGROUND: The requirement for subsequent cholecystectomy in patients with gallbladder in situ after endoscopic removal of stones from the common bile duct (CBD) is controversial. The aims of this study were to assess the requirement for subsequent cholecystectomy for gallbladder-related symptoms, and to identify the patients who develop symptoms after the endoscopic removal of CBD stones. METHODS: Of 241 patients with gallbladder in situ following endoscopic removal of stones from the CBD, 146 patients (78 men and 68 women; mean age 69 +/- 13 years, range 20-93) with a follow-up time of more than three months without elective cholecystectomy were enrolled in the study. Fifty-nine patients had gallbladder stones (single stones in 27 and multiple stones in 32) and 87 patients had gallbladder in situ without stones. The time from entry to the occurrences of death or cholecystectomy was evaluated retrospectively. Cox regression analysis was used to evaluate the risk factors associated with these events. RESULTS: The mean duration of follow-up was 24.1 +/- 18.0 months (range 3-70 months). During follow-up, seven patients (4.8%) underwent cholecystectomy, on average 18.4 months after CBD stone removal, as the result of acute cholecystitis in four cases, biliary pain in two cases and acute pancreatitis in one case. Laparoscopic cholecystectomy was performed in four patients and open cholecystectomy in three patients. Post-operative morbidity occurred in two patients, with improvement after conservative management. Nine patients (6.2%) died as the result of unrelated biliary disease. Age, sex, presence of gallbladder stones, multiplicity of gallbladder stones and underlying disease did not correlate with subsequent cholecystectomy by Cox regression analysis. CONCLUSION: Elective cholecystectomy is not warranted in patients with bile duct stones when the common duct can be cleared of stones by endoscopic sphincterotomy. We could not find any clinical predictors of further symptoms or complications arising from the retained gallbladder. PMID- 11855157 TI - Perception of wheezing in the elderly asthmatics. AB - BACKGROUND: In elderly asthmatics, underdiagnosis is one of the important features. The main reason for underdiagnosis is thought to be a low frequency in complaining of symptoms due to the reduction of intellectual recognition and physical activity. Among the various symptoms, wheezing is the principal clue in diagnosing bronchial asthma, and decreased complaints for wheezing are also noted in elderly asthmatics. The objective of this study is to determine if less complaints of wheezing in elderly asthmatic is due to decrease in the development of wheezing. METHODS: 61 young (20-39 years old), 68 middle-aged (40-59 years old) and 65 elderly (older than 60 years old) stable asthmatic subjects were studied (each group shall be called, hereafter, Young Group, Middle-aged Group and Old Group, respectively). During the methacholine induced airway narrowing, lung auscultation and questionnaire survey about presence and perception of wheezing were conducted in 194 asthmatics. RESULTS: One hundred and sixty-nine patients (87%) developed wheezing during the methacholine induced airway obstruction. The frequency of wheezing during the methacholine challenge was found to be comparable among the groups. The methacholine concentration, % fall in FEV1, and FEV1 levels of the initial detection of wheezing were not different among the groups. Among the patients who developed wheezing, 47 patients (77%), 42 patients (61.8%) and 26 patients (40%) complained of wheezing in Young, Middle and Old Group, respectively. CONCLUSION: In conclusion, the decreased perception of wheezing is a main factor for the low frequency of complaints of wheezing in elderly asthmatics. PMID- 11855158 TI - Total occlusion of left main coronary artery by dilated main pulmonary artery in a patient with severe pulmonary hypertension. AB - A 34-year-old woman was admitted to the hospital because of recently aggravated right heart failure without angina for 5 months. When she was 25 years old, patch repair with Polytetrafluoroethylene (PTFE) was performed for the secondum type of atrial septal defect (ASD) with moderate pulmonary hypertension. The chest PA, echocardiography and cardiac catheterization at current admission revealed Eisenmenger syndrome without intracardiac shunt. Chest CT scan with contrast revealed markedly dilated pulmonary trunk, both pulmonary arteries and concave disfigurement of the left side of the ascending aorta suggesting extrinsic compression, as well as total occlusion of the ostium of the left main coronary artery that was retrogradly filled with collateral circulation from the right coronary artery. The coronary angiography showed normal right coronary artery and the collaterals that come out from the conus branch to the mid-left anterior descending artery (LAD) and that from distal right coronary artery to the left circumflex artery (LCX) and to the distal LAD, respectively. On aortography, the left main coronary artery was not visualized with no stump, suggestive of total occlusion of the ostium of the left main coronary artery. From our experience, it is possible to say that the occlusion of the ostium of the left main coronary can be induced by the dilated pulmonary artery trunk due to ASD with pulmonary hypertension and that, if the ASD closure was too late, the narrowing or obstruction of the left coronary artery could not be resolved even after operation owing to irreversible pulmonary hypertension. PMID- 11855159 TI - A case of primary adenosquamous carcinoma of the liver presented with liver abscess. AB - Primary adenosquamous carcinoma of the liver is a very rare type of cholangiocarcinoma and is defined as a cancer containing both squamous and adenomatous components in the same lesion. Recently, we experienced a primary adenosquamous carcinoma of the liver presented as liver abscess. A 63-year-old man was presented with a 4-day history of fever and chill. The radiologic study showed a 4 cm-sized, central hypoattenuated mass with peripheral rim enhancement in the left lobe of the liver. Ultrasonography-guided aspiration and biopsy suggested an adenocarcinoma with abscess in the liver. At laparotomy, the tumor occupied the left lobe of the liver and invaded the right diaphragm. An extended left lobectomy and a partial excision of the involved diaphragm were done. Grossly, the tumor was 6 x 5 x 5 cm in size and had an eccentric necrosis. Microscopically, the tumor was composed of adenocarcinoma and squamous cell carcinoma with a transitional area. PMID- 11855160 TI - Retinal vein occlusion in two patients with primary antiphospholipid syndrome. AB - Primary antiphospholipid syndrome (APS) is a disease producing vascular thrombus with antiphospholipid antibody without association with autoimmune diseases as systemic lupus erythematosus. Retinal vein occlusion is a rare vascular manifestation in primary APS. We describe 2 cases of primary APS presenting with developing blurred vision. Each had central retinal vein occlusion and high titer of IgG anticardiolipin antibody. PMID- 11855161 TI - A case of mediastinal angiomyolipoma. AB - Angiomyolipoma is a common tumor of the kidney but has rarely been found in the mediastinum. We report a case of angiomyolipoma of the posterior mediastinum in a 62-year-old woman. She experienced exertional dyspnea and intermittent cough at admission. Computed tomography indicated a tumor located at the left paravertebral and upper posterior mediastinum and MRI imaging demonstrated a mass with low signal intensity in T1-weighted image at T4-5 level. Thoracotomy was done for surgical removal of the tumor and histologic examination revealed a mesenchymal tumor composed of mature fat, capillaries and smooth muscle fibers. The tumor was immunohistochemically positive for CD34 and factor-VIII (for vascular component) smooth muscle actin (for smooth muscle component) and S-100 protein (for fat component). There have been four case reports about mediastinal angiomyolipoma, namely three Japanese cases and one French case. It is suggested that angiomyolipoma could be considered for the differential diagnosis of mediastinal tumors. PMID- 11855162 TI - A case of hereditary angioedema associated with idiopathic hypoparathyroidism. AB - Hereditary angioedema is a rare autosomal dominant disease characterized by the edema of subcutaneous tissues, respiratory tract and bowel. It is caused by the deficiency of C1 esterase inhibitor. Hereditary angioedema may be associated with autoimmune diseases, such as systemic lupus erythematosus, rheumatoid arthritis, autoimmune thyroiditis and glomerulonephritis. We report a 34-year-old male patient with hereditary angioedema who developed idiopathic hypoparathyroidism. Autoimmunity seems to be an important basis of this association and it might be caused by the immune dysfunction due to decreased level of complements; nevertheless, a casual association could not be excluded. To our knowledge, this is the first report of hereditary angioedema in association with idiopathic hypoparathyroidism in the medical literature. PMID- 11855163 TI - Ensuring health-site quality. PMID- 11855164 TI - Mount Sinai NYU health: a portal experience. PMID- 11855165 TI - Marketing your site re-launch. PMID- 11855166 TI - Three ways to improve the site user's experience. PMID- 11855167 TI - Pew report: HIPAA privacy regulation leaves many unprotected. PMID- 11855168 TI - [Some issues on assessing natural capital of environment and ecosystems]. AB - Many researchers recently have paid attentions to analyzing ecosystem services and assessing natural capital stocks of environment and ecosystems. However, some works failed to make a distinction among the concepts, to evaluate the value and merit of ecosystems, and to integrate assessment approaches. This paper attempts to calibrate those deviations and to present new paradigm for monetary evaluation of ecosystem services and its capital stocks. Firstly, capital value of environment and ecosystems as well as species' value should be evaluated. Secondly, new comprehensive paradigm should be employed to assess the service value of environment and ecosystems, which integrates type and regional models. Thirdly, special attentions should be paid to patterns of natural capital of environment and ecosystems due to their spatial and temporal accumulations and flows. Finally, some suggestions are proposed for assessing natural capital. PMID- 11855169 TI - [An exponential universal formula and evaluation model of sustainable development for cities]. AB - In order to study an evaluation model of sustainable development for cities, analysis was performed to study the features of indicator system of community, economy and environment. Based on the genetic algorithms optimum, an exponential universal formula of single indicator was proposed. Evaluation model of sustainable development about the coordinative development of community, economy and environment was developed using weighting method combining the general contrast algorithm with analytic hierarchy process for different structure. The evaluation results of sustainable development for examples using this model showed that the exponential formula of single indicator based on parameters optimum and the evaluation model are not only simple and practical, but also comparisonable, realizable and general. PMID- 11855170 TI - [Quantitative structure and activity relationship for epoxides]. AB - Mutagenic activity data of 86 epoxides were collected from a CD system of Registry of Toxic Effects of Chemical Substances (RTECS 1998 Version). By using principal component analysis, the 5 kinds from 11 kinds of molecular structure descriptors, such as the number of carbon atom, the number of benzene-cycle, the number of hydrogen atom, the number of substituted alkyl group and the number of oxygen atom which can have influence on their mutagenic activity very much were chosen. After learning of above samples. The Back-Propagation Method(BP) network structure was optimized, the learning sets of sample were classified, then unknown samples were predicted one by one; for 86 samples, the correctly classified rate can reach to 92% between the low class and the high class. Results shown that the special molecular structure, the epoxides of polycyclic aromatic hydrocarbons contained 3-4 cycles, can be responsible for their very high mutagenic activities. PMID- 11855171 TI - [Supercritical and near-critical fluid solvents assisted reaction and separation processes]. AB - The tunability of supercritical and near-critical fluid (S/NCF) solvents offers environmental improvements and economic advantages from improved performances and flexibility for separation and reaction processes through density changes or cosolvents. The paper reviews the sustainable reaction and separation processes in S/NCF solvents such as supercritical carbon dioxide and near-critical water. PMID- 11855172 TI - [Values of carbon dioxide emission from different land-use patterns of alpine meadow]. AB - The value order of carbon dioxide emission from different land-use of alpine meadow was significantly with season changing. It was found that the Potentilla fruticosa shrub meadow(C, 1871.40 g/m2) > Kobresia humilis meadow(A, 1769.63 g/m2) > Degraded Potentilla fruticosa shrub meadow(D, 1495.60 g/m2) > Degraded Kobresia humilis meadow(B, 1191.26 g/m2) during growth season, A(661.46 g/m2) > C(550.90 g/m2) > B(502.50 g/m2) > D (384.50 g/m2) in rest period; and A(2431.09 g/m2) > C(2422.30 g/m2) > D(1880.10 g/m2) > B(1694.06 g/m2) for whole year, separately. This deference not only decided the soil microbial activity and soil characters, but also have the closely relationship to shortage of frozen period. PMID- 11855173 TI - [Development of biogenic VOC emissions inventory with high temporal and spatial resolution]. AB - A new method was developed to estimate biogenic VOC emissions with high temporal and spatial resolution by use of Mesoscale Meteorology Modeling System Version5 (MM5). In this method, the isoprene and monoterpene standard emission factors for some types of tree in China were given and the standard VOC emission factors and seasonally average densities of leaf biomass for all types of vegetation were determined. A biogenic VOC emissions inventory in South China was established which could meet the requirement of regional air quality modeling. Total biogenic VOC emissions in a typical summer day were estimated to be 1.12 x 10(4) metric tons in an area of 729 km x 729 km of South China. The results showed the temporal and spatial distributions of biogenic VOC emission rates in this area. The results also showed that the geographical distribution of biogenic VOC emission rates depended on vegetation types and their distributions and the diurnal variation mainly depended on the solar radiation and temperature. The uncertainties of estimating biogenic VOC emissions were also discussed. PMID- 11855174 TI - [Influence of planting density and precipitation on N2O emission from a winter wheat field]. AB - To investigate the impact of plant density on N2O emission from winter wheat field and the cause of seasonal variation in the emission, field experiment with four planting rates of 0, 90, 180 and 270 kg/ha was conducted at the Jiangning County near Nanjing during 1999-2000 winter wheat growing season. Data of the field measurements indicated that the N2O emission rates during the season from planting to overwintering were not influenced by the plant density, while the emission was positively correlated with the planting density during the season from turning green to maturity. The emissions from the field plots with planting rates of 0 and 90 kg/ha were not found to be significantly different. A further analysis suggested that the seasonal variation of N2O emission be mainly influenced by precipitation, which could be quantitatively described by an exponential function of a weighted average precipitation of 6-day period before measurement. PMID- 11855175 TI - [Nitrogen oxides emissions arising from commercial energy consumption in China]. AB - According to national and provincial energy consumption and specific NOx emission factors for different sectors and fuel types, NOx emissions resulted from commercial energy consumption in China in 1990's were presented. Specially, NOx emission inventories by provinces, sectors and fuel types in 1997 were developed and discussed. Total NOx emissions increased from 8.4 Mt in 1990 to 12.0 Mt in 1996. However, compared with the peak value in 1996, NOx emissions in 1997 and 1998 decreased by an amount of 0.34 and 0.82 Mt respectively. It can be seen remarkable imbalance of NOx emissions by fuel types, economic sectors and provinces: 3/4 of the totals came from coal burning; sectoral emissions were dominated by industry (39.56%), power (36.74%) and transportation (11.22%); NOx emissions in Hebei, Jiangsu, Liaoning, Shandong, Shanxi, Guangdong, Henan, Heilongjiang and Hubei province exceeded 0.5 Mt, while those in Qinghai, Ningxia and Hainan were lower than 0.1 Mt. Regions with highest NOx emission intensities (> 10 t.(km2.a)-1) were Shanghai, Tianjin and Beijing municipality. In sum, NOx emissions and pollution mainly concentrated in the populated and industrialized areas of China: the Eastern Central and Southeastern provinces. PMID- 11855176 TI - [A novel Fe/AC desulphurizer at low temperature]. AB - Activated coke was used to support Fe2O3(Fe/AC) for flue gas SO2 removal. Reaction conditions on DeSOx activity were investigated. The results show that Fe/AC had higher activity than AC or Fe2O3 at temperature of 120 degrees C-250 degrees C. H2SO4 and Fe2(SO4)3 were formed after Fe/AC sorbed SO2, H2O and O2 increased the amount of SO2 adsorption. Fe/AC derived from AC of higher BET surface area had higher DeSOx activity. Fe/AC was suitable to be used at GHSV below 800 L/(kg.h). PMID- 11855177 TI - [Effects of air staging with absorbents on trace metal during coal combustion]. AB - Staged combustion was carried out on laboratory-scale pulverized coal combustion with different absorbents. The experiment indicated staged combustion increased emission of submicron particles, which went against the control of trace elements, especially for those of high volatile elements, such as Cu, Ni. The thermodynamics calculation also indicate the transformation of trace metal was different with different atmosphere, suboxidized and reduced species were more easily formed under reduced condition. In both conditions, absorbents show a certain absorptive ability to trace metal, and different absorbent had different ability. For unstaged combustion, kaolinite was the best for Co, Cr and Ni; dolomite for Be, and CaO for Cu. But for under staged condition, HZ- dolomite was the best for Be, Cr and Ni; Kaolinite for Co and Cu. PMID- 11855178 TI - [Sources analysis and contribution identification of polycyclic aromatic hydrocarbons in indoor and outdoor air of Hangzhou]. AB - Twelve polycyclic aromatic hydrocarbons (PAHs) were measured in eight homes in Hangzhou during the summer and autumn in 1999. The sources of PAHs and the contributions of the sources to the total concentration of PAHs in the indoor air were identified by the combination of correlation analysis, factor analysis and multiple regression, and the equations between the concentrations of PAHs in indoor and outdoor air and factors were got. It was indicated that the factors of PAHs in the indoor air were domestic cuisine, the volatility of the mothball, cigarette smoke and heating, the waste gas from vehicles. In the smokers' home, cigarette smoke was the most important factor, and it contributed 25.8% of BaP to the indoor air of smokers' home. PMID- 11855179 TI - [Treatment of ion-exchange monosodium glutamate wastewater by yeast]. AB - Two strains of yeasts, Candida halophila and Rhodotorula glutinis, were acquired through screening from ion-exchange monosodium glutamate wastewater (IEMGW) which was characterized by its high contents of COD (40,690 mg/L), NH(4+)-N (16,914 mg/L) and SO(4)2- (18,000 mg/L). The mixture of the two species had a good COD removal performance for treating IEMGW in a pH rang of 4-9. The yeast technology was superior to the conventional activated sludge in terms of COD removal and COD decomposition rate, In a dilution rate range of wastewater from 1:1 to 1:9, a COD removal of 84.5% and an average COD decomposition rate of over 1.0 kg/(kg.d) were obtained under an initial pH of 4. PMID- 11855180 TI - [Autotrophic denitrification of groundwater by electrochemical process]. AB - An autotrophic denitrification process and its reactor for removing nitrate from groundwater was investigated in this paper. Using activated carbon fiber(ACF) as electrodes, the electrochemical reactions could produce hydrogen as the donor for autotrophic denitrification. In the process, nitrate was removed effectively and no accumulation of nitrite in the effluent. The results in this study proved that some key factors, such as the applied current density, flow rate of water, oxidation-reduction potential and the nitrate concentration in raw water, influenced the electrochemically denitrifying effect. When the concentration of NO3(-)-N was 28.8 mg/L in inlet water, the optimum applied density was 9 mA, the highest hydraulic load of the reactor was 35 ml/h, and the reactor performed a practical buffering capacity to pH. After applying electricity to the reactor for 1 h, the oxidation-reduction potential decreased to below--200 mA, so an adaptable reductive environment could be provided in the reactor for denitrification. PMID- 11855181 TI - [Treatment of 2-naphtholwastewater using air oxidation and ion exchange]. AB - The properties of 2-naphtholwastewater were studied in details in this paper, the wastewater was characterized by the strong acidity, high buffer capability and concentrated salt. Air oxidation and anion exchange were employed for the treatment of the wastewater. Under formal pH, room temperature and 1 BV/h rate, the result showed that the removal of COD reached to 97% and the recovery of sodium naphthalensulfonate to 98%. Besides, wastewater treated can be reused for washing the sodium naphthalensulfonate. PMID- 11855182 TI - [Extraction and stripping of H acid and DSD acid wastewater]. AB - H acid and DSD acid are two important substrate of dyes, which wastewater is treated more difficulty because of containing multi-component, with high chroma value and nonbiodegradability. A treatment way of this wastewater by extraction based on chemical association was carried out. Trialkylamine (Alamine 336) and chlorinated quaternary ammonium salt (Aliquat 336) were used as reacting agent with n-octanol, kerosene as modifier and diluent respectively. Results showed that the extraction efficiency depended on solution pH. Alamine 336 could be used to treat waste water with low pH while Aliquat 336 was high efficiency for waste water with pH > 6. Recovery yield of H acid and DSD acid was greater than 99% and the chroma value of wastewater was reduced effectively by simulating multi-stages cross-flow extraction. Back-extraction is quite easy and the solvent could be regenerated. PMID- 11855183 TI - [Non-filamentous activated sludge bulking caused by the deficiency of nitrogen]. AB - The effect of nitrogen deficiency on activated sludge bulking was studied specially in some experiments carried on a sequencing batch reactor fed with beer wastewater in this paper. The experimental results showed that the sludge settled properly at a influent BOD/N value of 100/4. When the value of BOD/N was 100/3 and 100/2 respectively, non-filamentous activated sludge bulking caused by an excessive growth of viscous Zoogloea with high moisture content occurred. When the value of influent BOD/N was 100/0.94, more serious non-filamentous bulking occurred. Furthermore, the effect of nitrogen deficiency on the nitrogen sources and phosphorus sources utilization rate and the COD removal rate was studied in the experiments. PMID- 11855184 TI - [Characteristics of nitrogen and phosphorus pollution in the middle and small creeks, suburban Shanghai]. AB - 1. The middle and small creeks in suburban Shanghai were under heavy eutropic condition, with high loadings of nitrogen, phosphorus and organic pollutants. KN, TP and CODCr of the surface water of them were several times greater than the critical values of Types V, the worst level of water. 2. Spatial variations of N and P pollution were observed, as the creeks in different places accepted different types and amounts of pollutants. Affected by domestic sewage, the creeks nearby residential areas were often heavily polluted by NH4+ and P. The creeks in agricultural fields were more easily polluted by NO3- than by NH4+ and phosphorus. The creeks nearby livestock farms often witnessed sudden soaring of NH4+ and P. 3. N and P loadings of the creeks were also affected by the seasonal evolvement of ecological environment. With the increase of temperature and biomass, NH4+ and NO3- in the surface water were largely consumed and decreased as a result; while organic N and particle P increased as the enhancement of suspending biological particles. NO2- in the surface water increased with temperature as nitrifying bacteria became active. Total P and water soluble P also showed the trend of enhancement with the intensification of biological activities, however, they were also controlled by the condition of pH and DO in the water. 4. At the height of the summer, the surface and bottom water of the creeks differed in N and P loadings. NH4+, soluble P and total reactive P in the bottom water were higher than those in the surface; while NO3- and NO2- in the surface were higher than those in the bottom. Such water-quality stratification inevitably resulted from the enhancement of phytoplankton in the surface water and intensified release of NH4+ and P from the sediments. In the autumn, as-the temperature decrease, such phenomenon declined gradually. PMID- 11855185 TI - [Effects of grazing and cultivating on emission of nitrous oxide, carbon dioxide and uptake of methane from grasslands]. AB - Measurement of N2O, CO2 and CH4 with the optimized closed chamber/GC technique in the typical grassland, which were influenced by human activities, grazing and cultivating first time in situ were carried out in Inner Mongolia of P. R. China from 1998 to 1999. It was found that wheat cropping can increase N2O production 3 times than the native, and changed a CO2 sink of Inner Mongolia Steppe to a significant source, increased efflux CO2(C) average 14.3 mg.(m2.h)-1 from Jun. to Dec. in 1998. CH4 uptake was highly increased if there was grazing lightly. There were no effects of wheat cropping on CH4 uptake and no significant results of grazing on N2O emission. Soil moisture and temperature were the key factors, other than human activities, which affect the patterns of seasonal variation of N2O and CO2 emission and CH4 uptake. Highly inter-annual variations, which resulted from the annual precipitation, of the exchange fluxes and seasonal variation patterns of N2O and CO2 emission and CH4 uptake there were. It was a fact that N2O and CO2 emission from the soils was inversely related to CH4 uptake. PMID- 11855186 TI - [Biosorption of heavy metals by bacteria isolated from activated sludge]. AB - In this paper, biosorption efficiency of heavy metals by bacteria isolated from activated sludge was tested. It was shown that Pseudomonas pseudoalcaligenes and Micrococcus luteus GC subgroup B had high biosorption capacities for Cu(II) and Pb(II) ions. The adsorption of the two ions on Pseudomonas pseudoalcaligenes and Micrococcus luteus GC subgroup B exhibited Langmuir adsorption behavior with regression coefficient of 0.99. The pH value was the most important effect factor on biosorption of metal ions. The biosorption kinetic curves of Cu(II) and Pb(II) ions by cells showed two processes: rapid sorption on cell surface and slowly longer-term transformation into cells. Cu(II) ion can be desorbed effectively from cells of Micrococcus luteus GC subgroup B. PMID- 11855187 TI - [Sewage treatment by the canal reactor with biomembrane]. AB - This paper studies that utilizing sewage canal (waterway) of city to treat sewage on bases of researching water pollution, hydrological data and topography of the canal. A new canal reactor of bacteria membrane with continuous operation was designed, made and studied in the laboratory. The operation results of the reactor show that the reactor could effectively treat the sewage of city. The average data of continuous operation during 3 months showed that the removal ratios of CODCr and BOD5 were 88% and 95% in the range of CODCr concentration 180 mg.L(-1)-450 mg.L-1 in the inlet, separately. The technology also has the advantage of low capital cost, low operation cost. This will create the condition to apply the way to practical living sewage treatment of canal. PMID- 11855189 TI - [Biodegradation of polycyclic aromatic hydrocarbons by a preponderant brevibacterium]. AB - A preponderant brevibacterium was isolated from the sludge contaminated by coke plantwaste water for a long time. The bacterium was effective on the degradation and transformation of anthracene, phenanthrene and pyrene. The degradation process can be accelerated by ferric ions. 10 hours after aeration, the removing rates of anthracene, phenanthrene and pyrene by the brevibacterium were about 75%, 87% and 62% respectively. 106 hours after aeration, the removing rates of total organic carbon (TOC) caused by anthracene, phenanthrene and pyrene were 50%, 90%, and 50% respectively. PMID- 11855188 TI - [Effects of calcium on biological characteristics of wheat seedlings under ultraviolet-B radiation stress]. AB - Under simulated ultraviolet-B radiation (UV-B, 280-320 nm), effect of calcium on biological characteristics of wheat seedlings was studied with pot culture experiment. The results showed that the growth indexes of wheat seedlings on ground and underground in calcium concentration 2 mmol/L treated group were obviously reduced than that of wheat seedlings in control treated group (the descending range of 8.1%-36.1%) under UV-B stress (0.35 W/m2, 6 h per day, for 7 day). And the decreasing degree of growth indexes (3.2%-28.6%) for wheat seedlings in calcium concentration 6 mmol/L treated group were lower than in calcium concentration 2 mmol/L treated group. The experiment proved that calcium can mitigate the damage of ultraviolet-B radiation on wheat seedlings, and this effect was related to that calcium affected physiological and biochemical characteristics of wheat seedlings. PMID- 11855190 TI - [Mechanism of contour hedgerow's control for non-point pollution in the semiarid region in northwest Hebei Province]. AB - Slope nutrient loss is an important type of non-point pollution. The mechanism of non-point pollution control by using plenty of natural observed data in Hilly Loess region in Northwest Hebei Province, as well as combating with the large scale artificial simulation rainfall in the field slope was analyzed. The study results show that the critical slope length was 10-15 m on the condition of rainstorm, which led to the generation of rill and made the erosion amount went up obviously. The study show that the nutrient loss in sediment was the main type of non-point pollution, the contour hedgerow the process of non-point pollution by controlling the nutrient loss in sediment, then the study got the erosion control model of the contour hedgerow. PMID- 11855191 TI - [Effect of toluene on urease activity of Lou soil]. AB - Urease activity affected by toluene studying results showed as follows: Toluene had little action on Jackbean urease activity, but can remarkably increase soil urease activity in lower toluene concentrations and shorter time, the increasing variation was 1.48-3.96 times than without toluene treatment in seven soil samples tested. The variation was less than 3.98% when sterilized soil samples adsorbed the Jackbean urease. It may be because toluene can kill soil microorganism, release urease within cell and subsequently urease fixed into soil organic-inorganic colloidal. Different soil pH and microbiology can lead to vary differently for urease activity. PMID- 11855192 TI - [Leaching and vertical transport of Zirconium-95 in soils]. AB - The leaching and vertical transport of Zirconium-95 in three representational soils (paddy soil, red-yellow earth and sea mud) in Zhejiang province and the influence of salinity on the activities of Zirconium-95 in sea mud were studied with soil-pole installation. The results show that: 1. The content of Zirconium 95 in collected leachate was low after passing through soil column with 250 ml, 500 ml, 750 ml leaching amount. The amount of Zirconium-95 in collected leachate of paddy soil, sea mud and red-yellow earth was about 5.33%-7.68%, 0.77%-1.32% and almost 0 of labelled amount respectively. And with increasing the total amount of leaching water, the content of Zirconium-95 in collected leachate varied indistinctively, which indicated that once Zirconium-95 was absorbed by soils, it was very difficult to desorb. 2. In paddy soil, red-yellow earth and sea mud, 45.99%-50.02%, 96.02%-97.16% and 95.94%-98.01% radioactivity of Zirconium-95 was concentrated in the 0.4 cm upper layer soils respectively, which illustrated that most of Zirconium-95 remained in the surface soils. 3. With the increase of salinity in leaching water, the amount of Zirconium-95 in surface soils (0-0.2 cm) presented obviously increased, which demonstrated that increasing salinity could raise the adsorption rate of Zirconium-95 in soils. PMID- 11855193 TI - [Bioreactor-landfill site for leaching solution treatment]. AB - The study utilizes the system combined bioreactor with landfill site to treat leachate. The results show that the system helped the degradation of the organic pollutants in the leachate to be divided into two-phases, the hydrolytic fermentation and acid-production phases mainly occurred in the landfill site, the acidification rate was 40%-50%, the methane-production chiefly occurred in the bioreactor(UASB). It treated the leachate significantly and benefits the collection and utilization of methane gas. Also, the system accelerates the process of degrading municipal solid waste and stabilizing landfill site. PMID- 11855194 TI - Occupational nurses in waste water treatment. PMID- 11855195 TI - Challenges of the working breastfeeding mother. Workplace solutions. AB - 1. The number of women who breastfeed their infants after returning to work is disappointingly low. Many women stop breastfeeding soon after they return to work because of lack of preparation and support. 2. Continued breastfeeding provides health and psychosocial benefits for both mother and infant. 3. The employer experiences a cost saving benefit by keeping the postpartum employee working and by keeping her infant healthy as a result of breastfeeding. 4. Occupational and environmental health nurses can help bridge the gap between the postpartum employee and her primary health care provider. One strategy is to develop a lactation program for the work setting as described in a stepwise approach. PMID- 11855196 TI - Work-life balance in an industrial setting. Focus on advocacy. AB - 1. Employers have a role in assisting workers to manage conflicting priorities of work and family. 2. Management support and sensitivity to family and home life responsibilities and flexible application of policies are believed to be key factors in promoting work-life balance in the workplace. 3. Occupational health nurses have a role in advocating for revised company policies to assist employees to achieve and maintain a work-life balance. The information provided in this article can be used to guide future evaluation for the reduction of barriers to employee work-life balance. PMID- 11855197 TI - Toward a model for effectiveness. What Alberta occupational health nurses think. AB - Effectiveness is difficult to define or measure, but is frequently associated with cost. A two phase study conducted with occupational health nurses in Alberta, Canada resulted in a beginning model for effectiveness. In 1997, Phase One of an exploratory descriptive study focused on physical assessment by occupational health nurses (N = 137) and perceptions of effectiveness in practice (n = 104). In 2001, Phase Two used focus groups (n = 7) to determine occupational health nurses' reactions to the preliminary analysis of questionnaire responses on effectiveness. The focus groups confirmed and expanded categories, and reconfigured the developing model. The model makes explicit the foundations, functions, relationships, and goals for effectiveness in occupational health nursing practice. The foundation includes registered nurse (RN) experience and baseline competence comprising occupational health nursing education, RN and occupational health nurse experience, and multidisciplinary knowledge. Ten specific functions and nine relationships describe the occupational health nursing specialty practice and promote achievement of five goals: balance, communication, continuing competence, leadership, and trust. The goal of balance needs articulation in the nursing literature. PMID- 11855198 TI - Outsourcing occupational health services. Critical elements. AB - Successful management of an outsourcing relationship produces a highly interactive, flexible relationship between two organizations. The unique skills and resources of the service provider can be leveraged by the purchasing organization to achieve its business goals. Occupational and environmental health nurses can orchestrate this process and implement this important management tool in the provision of quality occupational health services. PMID- 11855199 TI - Tips for shiftworkers. PMID- 11855200 TI - When disaster strikes. PMID- 11855201 TI - Ticking clock unites rivals. PMID- 11855202 TI - Electronic stepping stones. PMID- 11855203 TI - The HIPAA identifier rules. PMID- 11855204 TI - Paper documents get sharper image. PMID- 11855205 TI - Clinical trials getting a hand. PMID- 11855206 TI - No license to steal. PMID- 11855207 TI - Improving outcomes in rehabilitation. AB - This article addresses the issue of measurement in a geriatric rehabilitation program (GRP). The Sisters of Charity of Ottawa Health Service Geriatric Rehabilitation Program was found to have a long length of stay relative to peer programs. Therefore, a number of processes were adopted, over 15 months, to improve outcomes. The results of these interventions produced a net improvement in patient outcome including a threefold increase in rate of patient improvement and a corresponding increase in cost efficiency. PMID- 11855208 TI - The organizational structure of intensive care units and its influence on patient outcomes. AB - Despite the growing body of knowledge on the theory of organization, the application of such theory to the organization of intensive care units is in its infancy. Our knowledge about the influence of ICU organization on patient outcomes is limited. Development of instruments to measure ICU organization, and their implementation in studies of new therapies and technologies, will assist in demonstrating the effect of various models of ICU organization on the provision of clinical care. PMID- 11855209 TI - The realities of synthesizing and disseminating research evidence. AB - This article describes the results of a 15-month pilot project to determine whether producers and users of organizations' research findings can synthesize and disseminate evidence collaboratively. The findings of the project suggest that the collaborative approach is a feasible but costly endeavor owing to differences in need, expertise, experience and context. Funding agencies should consider alternative ways to build sustainable partnerships and to address the cultural and political contexts of organizations involved with the synthesis and dissemination of evidence. PMID- 11855211 TI - Adventures in research land: another glance "through the looking glass" to see what constitutes research. AB - There are many frames and rules of thumb for determining what constitutes 'research'. Some views are directed by the perspective or philosophical underpinnings of particular disciplines or approaches to research. Others are guided by rules concerning or definitions of what 'evidence' is, or whether 'new' knowledge is created. A recent paper by Jarvis suggests that we should differentiate among research, evaluation and measures to assure quality, and that this may help us steer a course through the roles and reasons for these various and varying activities. While the desire to clarify some distinct territory for what constitutes research versus something else is understandable, we argue that these distinctions in the end are at best unhelpful, can be misleading and actually do more harm than good--which in itself is an outcome that good research should avoid. This brief report is not a critique of the specific nomenclature suggested by Jarvis or other existing frames for identifying 'research'. Our intent is rather to begin a more general commentary on the very subject raised near the end of Jarvis's paper: "The three primary approaches to reviewing what we do are research, evaluation and quality assurance. There are similarities, differences and overlaps among these three approaches. They are part of a continuum with no clear distinctions between them." PMID- 11855210 TI - Public-private partnerships in healthcare: criteria for success. AB - Public-private partnerships present opportunities for healthcare providers to meet the dual challenges of shrinking public funding and increasing demand for care. Partnering with private sector organizations can provide new sources of capital, expertise and technology. Successful partnerships require specific skills and strategies. This article describes a pilot study that explored the literature and the experiences of participants in a small sample of interviews to compile a set of criteria for successful public-private healthcare partnerships. PMID- 11855212 TI - Our changing social values and healthcare: from social solidarity to consumerism? PMID- 11855214 TI - Of studies, summaries, synopses, and systems: the "4S" evolution of services for finding current best evidence. PMID- 11855213 TI - A journey to create a new patient-record system. AB - In fundamentally redesigning its patient-record system, West Park focused on the patient's story, streamlined the many chart forms, and integrated the documentation with a new care model that would bring the disciplines in closer collaboration. We involved stakeholders in the redesign process at every step, relying on leadership and collaboration throughout the organization. We sought to fit the new system to the organization--to "West Park it", and we continue to refine the new charting forms. PMID- 11855215 TI - Evidence-based psychiatric/mental health nursing. PMID- 11855217 TI - The invisible culture of the multiracial, multiethnic individual: a transcultural imperative. AB - The main purpose of this introductory article is to evoke professional awareness, spark interest, stimulate thought, and disseminate information concerning multiracial, multiethnic (multiple heritage) individuals within the United States. General background information, terms, definitions, ethno-historical influences, practice implications, and current issues will be highlighted. Areas for further exploration will be proposed. Transcultural imperatives urge all nurses and other health care professionals to become active participants in the new cultural evolution of a different, broader worldview that uncovers the "invisible" culture of multiple heritage individuals. The new vision challenges nurse and other health care professionals to embark upon a new journey in the quest for cultural congruent care for all individuals. PMID- 11855218 TI - Systematic assessment and evaluation of diversity content presented in classroom lectures: the FRDC tool. AB - The Fair Representation of Diversity Content (FRDC) Tool was developed to systematically assess and evaluate both qualitative and quantitative dimensions of diversity content incorporated into lectures in an undergraduate nursing course addressing basic nursing concepts and has relevance for other disciplines as well as Nursing. PMID- 11855216 TI - A community health promotion partnership model: the South Carolina health connection. AB - Over a five year period, the South Carolina Health Connection Project has evolved to multi-site, multi-organization community-base collaborative initiative. From this project over $60,000.00 in funds have been secured. However, when costing the human resources and many other in-kind contributions involved in the SCHC Projects activities, the Project can modestly be valued at nearly $200,000.00. The efforts of a few have been shared with others, who also shared the resources with others, and the health promotion empowerment cycle continues. We believe the South Carolina Health Connection is an exemplary of a Community Health Promotion Partnership Model. We hope you will agree! PMID- 11855220 TI - Are we our own impending threat? PMID- 11855223 TI - Sixth annual conference on infectious diseases focuses on bioterrorism, as well as more familiar problems. Monday, Dec 3, to Wednesday, Dec 5, 2001. AB - The Sixth Annual Conference on Infectious Diseases provided attendees with an opportunity to hear presentations by leaders in the field of infectious diseases and epidemiology who provided evidence that supports changing practices. Information supports the need to maintain well-known practices and recognize appropriate changes that will prevent undue patient and health care worker risk. PMID- 11855219 TI - Urban stress and mental health among African-American youth: assessing the link between exposure to violence, problem behavior, and coping strategies. AB - This project examines gender differences in exposure to violence, coping strategies and problem behavior among 306 African-American middle and high school students in the state of Virginia. Gender differences in problem behaviors among youth exposed to violence as either victims or witnesses are examined in addition to variations in coping strategies. Relying on recent research examining violent behavior and victimization events, the study focuses on the internalizing and externalizing behavioral characteristics (i.e., academic achievement, anxiety, depression, negative self-esteem, and delinquency) of urban students exposed to violence and the extent to which coping strategies differ. Results show specific gender differences with regard to problem behavior and coping strategy among African-American youth exposed to violence. For adolescent males, exposure to violence and victimization is strongly associated with externalizing problem behaviors such as delinquency, while adolescent females exposed to violence and victimization are more likely to exhibit internalizing symptoms indicative of post-traumatic stress disorder. Among students exposed to violence and victimization, females are more likely to use problem-focused coping (i.e., social support) as an adaptive strategy in comparison to males. Implications for intervention and future research are discussed. PMID- 11855224 TI - Optimize patient health by treating literacy and language barriers. AB - More than 90 million Americans have limited literacy skills. Almost two million US residents cannot speak English, and millions more speak it poorly. The stigma of illiteracy or the inability to speak a country's predominant language keep patients from disclosing their limitations. Recognizing these facts is an important first step in improving health education for this vulnerable population. By adapting teaching techniques to patients' special needs, nurses can ensure that patients understand their health problems and plan of care. Statistics dramatically demonstrate the high cost of neglecting these needs. Patients who do not understand their plan of care do not comply with instructions and, therefore, suffer unnecessary complications. Health care providers who can communicate with their patients through multilingual, low literacy patient education materials and with the use of qualified interpreters markedly improve the quality of care for their patients and the resulting outcomes. PMID- 11855225 TI - Caring for a limited-English proficient patient. AB - Each day in the United States, health care workers try to communicate with patients who are deaf, hard of hearing, or limited-English proficient (LEP). According to government regulations, these patients are guaranteed access to language accommodations. The legal implications of these regulations will be discussed along with ways to facilitate communication with patients who are deaf, hard of hearing, or LEP and the requirements for those who act as interpreters for such patients. PMID- 11855226 TI - Carotid artery stenosis and endarterectomy. AB - This article reviews the disease process of carotid artery stenosis, its symptomatology, and prognosis for progression to ischemic stroke. Indications for both medical and surgical treatment are reviewed as is criteria for surgical categorization as specified by the American Heart Association guidelines for carotid endarterectomy. Although diagnostic testing, medical treatment and alternative therapy options of angioplasty, stenting and thrombolysis are presented, a thorough overview of the surgical procedure of carotid endarterectomy is the major focus of discussion, as it remains the "gold standard" of treatment for severe carotid artery stenosis. PMID- 11855227 TI - Experiences of Australian Army theatre nurses. AB - As battles have raged throughout the centuries, nurses have cared for ill and wounded soldiers. One nursing role during war is theatre (i.e., OR) nursing. This article describes the role of Australian Army theatre nurses during the Vietnam War. It is based on information collected in a study of the experiences of Australian Army nurses who worked in operating theatres in Vietnam between 1967 and 1971. As nurses today focus on the future to find new ways to meet the demands of nursing ahead, it is important to reflect on the past, as they can learn from history and from other nurses' experiences. PMID- 11855228 TI - Using the Internet as an educational tool in the OR. PMID- 11855229 TI - Accident prevention in surgical settings--keeping patients safe. PMID- 11855230 TI - Recommended practices for use of the pneumatic tourniquet. PMID- 11855231 TI - Surgery for cancer patients. Critical care needs. AB - Patients with cancer provide a unique challenge for critical care nurses. Outcomes of clinical trials, new surgical modalities, and pioneering cancer treatments have helped prolong lives. The patient with cancer can require surgical and critical care intervention at the time of diagnosis, at the point of definitive therapy, or in the later stages of disease. Treatment is individual and sometimes aggressive. Surgical intervention is undertaken with a view toward the patient's ultimate outcome, quality of life, and potential cure. Multimodality cancer therapy will continue to flourish, demanding astute assessment skills by the critical care nurse. The critical care nurse must be able to integrate knowledge of the patient's type of cancer, treatment history, comorbid conditions, and surgical interventions into routine postoperative critical care. PMID- 11855232 TI - Selected neurologic complications in the patient with cancer. Brain metastases and spinal cord compression. AB - Nurses play a key role in care of patients with brain metastases and spinal cord compression. Care of this patient population is unique because the skills and knowledge of medical-surgical, neurologic, oncologic, and critical care nurses are required. With the wide variability among patients, a highly individualized care plan is necessary to meet the tremendous challenges of caring for patients with metastatic central nervous system disease. PMID- 11855233 TI - Pulmonary toxicities of cancer therapy. AB - Toxicity of the pulmonary region in persons with cancer can have singular or multidimensional causes. The seven distinct pulmonary toxicities are embolism, edema, pleural effusion, fibrosis, alveolitis, bronchiolitis obliterans, and radiation-induced pneumonitis. Although difficult to diagnose, all of these toxicities can be minimized by early assessment and immediate interventions. Astute nursing assessments and comprehensive treatment can make a difference in quality and quantity of life. PMID- 11855234 TI - Glycemic crises in patients with hematologic malignancies. AB - Persons with hematologic malignancies such as leukemia, lymphoma, or myeloma often have coexisting medical conditions. Among these may be diabetes mellitus. The physiologic and psychologic stress of diagnosis and treatment may precipitate the life-threatening complications of DKA or HHNS in this group of patients. People with personal risk factors may develop diabetes mellitus secondary to diagnosis and treatment and present with either DKA or HHNS. It is essential that the health care team have a heightened awareness of potential complications. These are complex syndromes involving severe hyperglycemia, metabolic acidosis, fluid and electrolyte imbalances, and neurologic and cardiovascular collapse. Working collaboratively with the critical care team to provide optimal care, nurses play an essential role in the management of these challenging complications of diabetes mellitus. PMID- 11855235 TI - Liver metastases. A case example. AB - The development of liver metastases is a potentially life-threatening event for cancer patients. Although the clinical presentation, diagnostic workup, and treatment can vary, each patient with liver metastases presents a unique challenge for the oncology nurse. PMID- 11855236 TI - Immunosuppression in transplantation. A new millennium in care. AB - Organ transplantation has grown tremendously during the last part of the 20th century. Cyclosporine, which was introduced almost 20 years ago, revolutionized transplantation as a viable treatment for end-organ failure. Successful transplantation outcomes have increased with the use of new and improved immunosuppression agents. The 21st century promises new challenges and discoveries as transplantation advances. PMID- 11855237 TI - Opportunistic fungal infections in the critically ill. AB - Opportunistic fungal infections are increasingly common in acute care and now represent 10% to 15% of all nosocomial infections. Few references and clinical resources are readily available for nurses in planning care for these patients (as opposed to bacterial diseases, which are better known). Nurses must recognize patients at high risk for fungal infections and develop more detailed assessment plans that include orifice and breath sound assessments. They also must be familiar with antifungal therapies and the specialized nursing care required to administer them safely and with the best bioavailability. PMID- 11855238 TI - Management of disseminated intravascular coagulation. AB - Disseminated intravascular coagulation (DIC) is a complex condition in which diffuse clotting and profuse hemorrhaging occur simultaneously. It is a serious, often fatal, condition that is estimated to occur in approximately 10% of all patients diagnosed with cancer. No single laboratory test is available that is absolutely diagnostic for DIC; however, several laboratory results, combined with certain clinical findings, will support the diagnosis. Despite the lack of a standardized protocol for clinical management, aggressive medical and nursing care can play a prominent role in the clinical outcome of the patient with DIC. PMID- 11855239 TI - Hemorrhagic disorders associated with thrombolytic therapy. AB - Thrombolytic treatment for AMI, acute ischemic stroke, and massive pulmonary embolism has shown significant benefit. Along with the potential increase in perfusion and decrease in cell death, however, comes potential complications. Bleeding is the most common complication associated with thrombolytic therapy regardless of the particular agent and can range from minor bleeding from an i.v. site to a life-threatening hemorrhage, such as GI bleeding. Expert assessment and management of patients who develop bleeding complications is critical to desired patient outcomes. Knowledge of the diagnosis for which a thrombolytic agent is used, pathophysiology, hemodynamic changes, and symptoms of complications associated with bleeding, all present a challenge to nurses. Research-based algorithms, protocols, or standardized treatment plans and a multidisciplinary approach to thrombolytic therapy provide the best opportunity for success, reducing the risk and enhancing early intervention of complications. Although thrombolytic therapy creates its own set of challenges, the alternative--failure to restore perfusion to the myocardium, brain, or pulmonary vasculature--presents both a different set of challenges and a dismal outcome. PMID- 11855240 TI - Ethical perspectives of reimbursement under economic pressures. AB - Health care is a primary need and an important goal for individuals and for society overall. From an ethical perspective, resources should be allocated appropriately so everyone can enjoy the best health care possible. With limited resources available from an economic reimbursement perspective, we need to analyze the philosophy and values that are important to us to make ethical decisions. This article describes key values and philosophies and concerns about societal attempts to reform the system. PMID- 11855241 TI - Advance directives in critically ill cancer patients. AB - Although advance directives have been used since the 1970s, the discussion of these documents has become more common since the enactment of the Patient Self Determination Act in 1991. This study evaluated the frequency of advance directives in critically ill patients at a tertiary cancer center and found that cancer patients had a relatively low completion rate of advance directives (27%). The finding that advance directives were more common among Caucasians than other ethnic groups signifies the importance of considering cultural differences when addressing end-of-life issues with patients. The documents also were found more often in older patients with serious diseases. The relationship between hospital mortality and advance directives is complex. The most seriously ill patients and patients with leukemia had the highest mortality and the highest rate of advance directives. Further research on the interactions among mortality, advance directives, and severity of illness is needed. This research contributes to the body of knowledge available on advance directives and particularly sheds light on advance directives in critical ill cancer patients. PMID- 11855242 TI - The influence of do-not-resuscitate orders on care provided for patients in the surgical intensive care unit of a cancer center. AB - Results of this study indicate that critically ill cancer patients who had a DNR order written while in the SICU received less medical intervention after the order was executed, whereas supportive nursing care remained unchanged. Addressing DNR issues in cancer patients earlier during their hospital course or even as outpatients may better serve patient and family wishes and reduce unnecessary costs of futile care. PMID- 11855243 TI - Incorporating critical care oncology nursing in a baccalaureate of science in nursing curriculum. AB - Undergraduate nursing curricula traditionally provide a superficial overview of oncology with only a passing mention of critical care oncology concepts. With an unprecedented nursing shortage predicted in the coming decade and acute care facilities likely to overflow with an elderly, chronically ill patient population, nursing education programs must strive to assure that nursing curriculum remains relevant in the face of these future challenges and changes in health care. Rather than require employers to provide the oncology education needed by the nursing staff during the orientation period, undergraduate nursing programs must accept the commitment to provide a stronger oncology and critical care nursing knowledge base in the basic nursing curriculum. This article describes how the baccalaureate degree nursing program at Houston Baptist University accepted this challenge and incorporated content on the management of oncologic emergencies within an existing critical care nursing course. Discussion of the development of the oncology content and the benefits of incorporating the content within a critical care nursing framework are also provided. PMID- 11855244 TI - Altered cellular anatomy and physiology of acute brain injury and spinal cord injury. AB - The cellular elements and the chemical mediators in secondary injury following TBI or SCI do not act alone. The interconnections between the cellular elements and their secretions, the immune system, and the nervous system are highly regulated in normal physiology, which benefits the organism. When there is traumatic injury to cells in the CNS, the interconnections between the systems become more than tight; these systems act together to strangulate the tissue, depriving it of local control over microcirculation and necessary oxygen, rendering membrane potentials useless to modulate neuron function. Surgical intervention during the acute stages of TBI and SCI continues to advance as do biomechanical and bioelectric therapeutics during the chronic and rehabilitation stages. There is hope, too, for effective pharmacologic intervention at the initial stages, before secondary injury begins. The fact that the mediators of secondary injury are already resident in normal physiology, whether during development or adulthood, means that their activity can be modified by specific agonists and antagonists directed to restoring homeostasis or promoting pathways that can lead to regeneration. This is the direction of much current basic and clinical research. PMID- 11855245 TI - Keeping the brain in the zone. Applying the severe head injury guidelines to practice. AB - Providing care to the TBI patient population with severe injuries requires an integrated multidisciplinary approach. The team in clinical practice must be willing to examine its own practice, seek out the latest information on TBI, and critically analyze the information. Members must be open to changing their own practice when the data presented support change. Interventions based on scientific evidence provide a strong foundation for delivering care. The standardization of these interventions into protocols facilitates team communication and coordination. Measuring outcomes is imperative for evaluating the effectiveness of current treatment algorithms. Changes in treatment practice should be based on the measured outcomes and advances in the scientific literature. PMID- 11855246 TI - Intracranial pressure monitoring and assessing intracranial compliance in brain injury. AB - Caring for the patient with a brain injury is a dynamic process with the goal of providing therapy to prevent secondary injury. Until practitioners have a better understanding of the pathophysiology of ischemia and the response of therapies for treating increased ICP, they must use the tools that exist. ICP monitoring gives a rough index of the relationships and the response of the intracranial contents to changes in volume that may produce increases in pressure and further damage. Understanding the information supplied by ICP monitoring is imperative to successful management of increased ICP. PMID- 11855247 TI - Microsensor and microdialysis technology. Advanced techniques in the management of severe head injury. AB - Neuroscientists continue the search for the "magic bullet" that will prevent the deleterious effects of primary and secondary brain injury. Indirect measurement of the effects of primary and secondary brain injury through the study of ICP- or CPP-directed management, CBF monitoring, Sjo2 monitoring, and TCD monitoring has led to improved care of persons with brain injury. Although the findings from brain injury research using microsensor and microdialysis technology are only preliminary and extensive research is still needed, these technologies have dramatically expanded knowledge about brain injury at the cellular level. Extended neuromonitoring is poised to enter a new and exciting phase because of the growth in knowledge regarding the cellular events associated with brain injury. The recent approval of NeuroTrend by the FDA will further promote this growth. Applications of the technology have already expanded to include uses beyond the management of traumatic brain injury. Microsensor and microdialysis technology is being used intraoperatively to determine "safe" temporary clipping times for aneurysm surgery and is also being used within the critical care setting to improve the monitoring and management of subarachnoid hemorrhage patients who are experiencing vasospasm. The ultimate application of this new technology is to improve long-term outcomes for patients with brain injury through the reduction of secondary brain injury. If that goal is to be accomplished, then it will be important for nurses caring for patients with brain injury to become immersed in this exciting new phase in brain injury monitoring. Nurses must obtain a comprehensive knowledge base of brain injury pathophysiology and how extended neuromonitoring can lead to improved outcomes. Technical proficiency will also be important to ensure that treatment and research conclusions are based on accurate data. Finally and perhaps most importantly, it will be critical for nurses to participate in and develop research studies that explore the impact of interventions, especially nursing care activities, on the injured brain if these exciting new advances are to be translated into tangible benefits for brain-injured patients. PMID- 11855249 TI - Pulmonary challenges in neurotrauma. AB - Traumatic injury to the central nervous system presents a formidable clinical challenge to any practitioner. Part of that challenge is to anticipate the pulmonary sequelae that may occur in this patient population. Direct pulmonary trauma commonly occurs with CNS injury and requires immediate treatment to prevent further compromise of the patient's condition. Neurologic deficits can then be compounded by complications such as ARDS and aspiration pneumonia that can be minimized if not prevented. The nurse is a key health care professional who can implement these interventions and effect better patient outcomes. PMID- 11855248 TI - Cognitive impairments following traumatic brain injury. Etiologies and interventions. AB - Brain injury is a dynamic process that continues for weeks. Recovery is also a lengthy process, proceeding in overlapping stages along with injury. The outcome for many patients with TBI is an inability to fully participate in life events because of cognitive impairments. Physiologic responses throughout the injury and recovery are punctuated by neuroprotective and neuroplastic events. The time course of these injury and recovery activities requires that medical and nursing therapies are targeted across the trajectory of injury as damage and recovery processes are occurring. Prevention of secondary injury using medical and nursing strategies should be of paramount importance. Altering the environment by providing meaningful yet novel sensory stimulation may enhance plasticity and lead to reorganization of structures that support cognitive processes. Administration of neuroprotective agents in an effort to control damage from neurochemical processes should proceed as these agents become approved for clinical use. Active participation in rehabilitation programs and neuropsychologic testing provide additional avenues for identifying and addressing cognitive impairments. The complex relationship between injury and cognitive impairment is slowly being unraveled. Through an understanding of the brain structures and networks associated with information processing as well as the pathophysiologic consequences of brain injury, critical care nurses can design evidence-based regimens of care that preserve cognitive function and result in improvement of long-term cognitive outcomes and fuller participation in everyday life activities. PMID- 11855250 TI - Nutrition strategies in neurotrauma. AB - A basic understanding of metabolic alterations that occur following neurotrauma is essential for addressing nutritional requirements. Interventions must be research based and must focus on the support of metabolic alterations, minimizing the effect of catabolism and optimizing caloric delivery to meet metabolic demand. The goal of accuracy in the delivery of nutritional support is to ensure a reduction in patient morbidity. Nutritional support requires an ongoing, daily assessment of caloric goals, protein requirements, patient responses, and assessment of nutritional laboratory values. Using this strategy, neurotrauma patients will have the greatest opportunity for a positive outcome. PMID- 11855251 TI - Penetrating trauma to the head. AB - Penetrating trauma to the brain is not as common as blunt trauma; however, the incidence is becoming a frequent occurrence in our society. Rapid transport to trauma centers where definitive care can be rendered is essential. Outcome depends on the site of the missile tract, the presenting neurologic status, and the extent of neurologic tissue destruction. Neurologic deterioration occurs rapidly, and outcome results seem to depend on the patient's neurologic status at the time of surgery. CT scanning is the diagnostic procedure of choice and should be performed if the patient's condition is stable (see Fig. 3). Aggressive removal of missile and bone fragments needs to be balanced by the knowledge that it is preferable to leave behind a few hard-to-reach fragments than to increase the patient's neurologic deficit. CT scanning in the postoperative period is very helpful in identifying abscess formation as well as new or recurrent hematomas, edema, and areas of tissue injury not evident at the time of initial scanning. Antibiotic therapy should be initiated preoperatively. Control of elevated ICP plays a significant role in decreasing mortality and morbidity. Judicious debridement of injured brain combined with medical management of increased ICP will maximize the quality of recovery and increase survivability. Although great strides have been made in reducing mortality and morbidity for trauma patients, the sad issue is that the majority of traumas are preventable. Until society is willing to understand that it needs to make firearm safety a priority, there will always be patients to care for who have sustained a penetrating injury. PMID- 11855253 TI - Acute spinal cord injury. Nursing considerations for the first 72 hours. AB - The effects of spinal cord injury can be devastating. An understanding of spinal anatomy and the mechanisms of injury provide the basis for correlating presenting signs and symptoms with the extent of neural damage. Understanding the principles of acute care management will help guide the nurse in executing interventions aimed at preservation of neurologic functions. Research must continue to focus on quality-of-life issues and methods of preventing both primary and secondary injuries. PMID- 11855252 TI - Neurotrauma in infants. Shaken impact syndrome (inflicted head injury). AB - In summary, inflicted cerebral trauma is a devastating but ultimately preventable brain injury caused by unique physiologic mechanisms that result in a distinctive pattern of injury. Nurses in a variety of settings, from the clinic and emergency room to the pediatric intensive care unit, play a major role in the identification, documentation, diagnosis, treatment, and prevention of ICT. The nurse's unique training also promotes prevention education and nonjudgmental support of the family. PMID- 11855254 TI - Spinal cord injury in the elderly. AB - Caring for patients who have sustained an SCI requires the nurse to be knowledgeable in the pathophysiology of the disorder, the sequelae following the injury, and the nursing care activities. Additional knowledge is required of the nurse when the patient with SCI is elderly. The nurse must be familiar with the changes in the body systems caused by aging, the alterations in the spine from aging, and how these changes impact the sequelae and nursing care activities. PMID- 11855255 TI - Use of an acute spinal cord injury clinical pathway. AB - Phased outcome clinical pathways can be useful in the management of neurotrauma patients in the acute care setting. By developing multidisciplinary plans of care that focus on patient and family outcomes and not arbitrary points in time, hospitals can provide quality care to trauma patients that is both appropriate and cost effective. In fact, this type of plan for SCIs can be expanded on and used across the health care continuum from prehospital to community reintegration. Providing collaborative quality care will result in improved outcomes for both patients and health care institutions. PMID- 11855256 TI - Brain death and organ donation. AB - Critical care nurses are essential team members during the process of determining brain death and preparing for organ donation. Using their knowledge of the criteria for brain death, they care for the dying patient, support the grieving family, and participate in the consent process for organ donation. Nurses make a critical difference in saving the lives of others through the gift of life. PMID- 11855257 TI - Understanding culture and ethnicity: basic rudiments of an "anthropsychological" perspective for understanding human behavior. AB - While there appears to be general consensus in the behavioral sciences as to the importance of culture and ethnicity in shaping behavior, the discipline of psychology, a major producer of psychotherapists, is often resistant to the study of the basic concepts; seeing it as the domain of other disciplines. Thus, psychologically trained practitioners may not have rudimentary understanding of these concept; while endorsing and even trying to incorporate ethnically and culturally sensitive practices and techniques. This paper provides a basic reformulation of culture and ethnicity and presents the novel concept of an "anthropsychological" model for understanding individual behavior while being sensitive to ethnicity and culture. PMID- 11855258 TI - Organizational culture and African American nurse faculty productivity: a comparative study. AB - The primary purposes of this study were: (a) to examine the relationships among factors associated with organizational culture and the scholarly productivity of African American women nurse faculty teaching at historically black (HBCUs) and predominantly white colleges and universities (PWCUs) and, (b) to compare the differences in African American women nurse faculty productivity at HBCUs and PWCUs. Data were collected using a two-part survey, mailed to the campus addresses of 467 African American nurse faculty. A 31.5% response rate (N = 147) was achieved. A variety of scales using multiple statistical techniques were employed to analyze the data sets. PMID- 11855259 TI - Cultural considerations for treatment of childhood obesity. AB - Childhood obesity has become one of the most common health problems facing children in America. Results from the Third National Health and Nutrition Examination Survey reveal that ethnic minority children in the United States are at particular risk for development of cardiovascular disease due to their disproportionate levels of obesity. In treating childhood obesity among ethnic minorities, practitioners need to be mindful of the cultural norms surrounding body size. Additional concerns that must be addressed include the effects of target marketing of unhealthy foods toward ethnic minorities and environmental deterrents to outside physical activities, to name a few. Strategies given to address the problem of childhood obesity among ethnic minorities include, increasing the child's physical activity, reducing television viewing and the adoption and maintenance of healthy lifestyle practices for the entire family. PMID- 11855260 TI - Financing Estonia's unemployment insurance system: problems and prospects. AB - In recent years, favorable media coverage of the glowing employment situation in the United States has been the norm. History, for one, won't permit us, however, to become complacent with what appears on the books to be a rosy economic picture for the nation. Moreover, the mounting recognition that the U.S. economy is inextricably tied to those of other nations--large and small--serves to keep us vigilant. This paper allows us a comparative exploration of the employment conditions in Estonia and the problems and prospects of financing its unemployment insurance program. PMID- 11855261 TI - Israeli families immigration and intercultural issues: challenges to mental health counselors. AB - Israeli families who immigrate to the United States present certain sociocultural issues that need to be recognized in order to provide culturally relevant, respectful and empowering mental health counseling. Israeli families are further situated within the complex social, cultural, economic and political context of Jews in the United States, who are themselves rarely recognized as a minority group. The purpose of this article is to present some major themes observed in working with Israeli families. A brief case example and analysis are presented, along with some general guidelines for counselors working with Israeli families. PMID- 11855262 TI - The meaning of racism when the 'field" is the other side of town. AB - When a researcher who belongs to the dominant white western society engages in research with members of a nondominant racial group, she often experiences a profound transition into understanding the meaning of personal and societal racism. What happens when the fieldwork is on the other side of town and the researcher must move back and forth between a middle class white world and a poor black neighborhood? As the researcher becomes more involved with her research group, the meaning of "home" becomes lost as the researcher inevitably struggles with the significance of white privilege and the consequences of the racism this privilege serves to perpetuate. PMID- 11855263 TI - Rehabilitation and rehabilitation nursing. AB - This article provides some of the foundational content that forms the basis of rehabilitation and rehabilitation nursing. The content included definitions, rehabilitation philosophy, and rehabilitation teams. A particular case example was integrated throughout the article to exemplify the content. PMID- 11855264 TI - Ethical issues from critical care to rehabilitation. A challenge for specialty nurses. PMID- 11855265 TI - Survivorship. Living through a life-altering event. AB - Survivors living with a life-altering event consider themselves to be ordinary people dealing with extraordinary situations. As survivors live through the continuum of events, issues emerge in the physical, psychologic, social, spiritual, and financial domains. Managing symptoms, accepting changes in functional status, confronting loss of control, sustaining social support, probing for the meaning of events, and maintaining affordable health insurance become daily challenges for survivors and their families. Through perseverance, survivors conquer the crisis, realizing personal strengths and abilities as a result of the conquest. Nurses play a pivotal role in assisting survivors and their families in learning how to live. PMID- 11855266 TI - Therapeutic touch for a patient after a Whipple procedure. AB - TT is a healing modality considered useful for nurses who wish to develop nonpharmacologic methods to reduce pain and anxiety in their patients. TT should be considered an adjunct to pain medications and should be used in that context. In the stressful environment of ICU, the critical care nurse should pursue any means to provide decreased anxiety and pain and increased comfort. The patient in this article experienced pain relief and comfort with the use of TT. The Whipple procedure was performed on the patient, yet with the use of TT he was able to heal rapidly and was discharged ahead of schedule. No studies have been conducted on the use of TT to shorten hospital stays, yet that would be an interesting outcome to address in future studies. PMID- 11855267 TI - Rehabilitation and iatrogenic complications of critical care. PMID- 11855268 TI - Pulmonary rehabilitation after acute COPD exacerbation. AB - Acute COPD exacerbation provides a window of opportunity for providing teaching, supportive care, and rehabilitation to a group of patients at high risk for complications and re-admissions. This article has attempted to make a case for the unique needs of this group of patients. Treatment of exacerbation is often viewed as "routine" by hospital staff and generally has focused upon the pharmacologic regimen of oxygen therapy, bronchodilators, antibiotics, and steroids. Within a few days, the patient is usually less dyspneic and is discharged with a combination of MDIs and a plan for steroid taper. All too often, the patient returns home with no idea of how to regain lost function or prevent future exacerbations. The use of the MAC provides a more complete picture of the unique needs and capabilities of COPD patients and brings together the collective expertise of the entire health team in designing a plan of rehabilitative care. Planned monitoring of the patient after discharge is an integral component to the success of the rehabilitation approach and the ability of the patient and family to move toward self-management of this chronic disease. Transitional care programs, community nurse referrals, telephone contacts, and outpatient visits are all appropriate methods for providing the type of support required to sustain rehabilitation. Traditionally, nurse researchers have examined psychosocial and functional performance outcomes of rehabilitation, however, nurses are in an ideal position to explore the methods of rehabilitation that yield the most positive outcomes over time and are sustainable over the course of the disease. Nursing can and should assume a leadership role in such initiatives. PMID- 11855269 TI - Living the healthy heart path. Rehabilitation after a cardiac event. AB - Although cardiac rehabilitation has traditionally been linked to specific programs in which patients participate after having a cardiac event, the concept of cardiac rehabilitation has expanded to many domains of nursing. Not only are critical care nurses the initiators of cardiac rehabilitation through thorough and accurate teaching of the cardiac event but all nurses should be attuned to the modifiable risk factors to direct patients at high risk for possible prevention of a cardiac event. It is a critical care nurse who is knowledgeable about the focus and goals of cardiac rehabilitation, who incorporates the patient's family in the early rehabilitation stages, and who considers cultural variations who can provide optimal care to the patient, helping him or her lead a healthy and productive life. PMID- 11855270 TI - Advances in spinal cord injury care. PMID- 11855271 TI - Rehabilitation after a hip fracture. Special needs of the elderly. AB - Hip fractures are one of the most common and potentially devastating injuries in the geriatric population. The incidence, morbidity, and health care costs associated with hip fracture among older persons are well recognized. Because of the complex health care needs of the elderly, rehabilitation after a hip fracture can present a challenge at a time when the nursing workforce and health care funding are declining. Aggressive rehabilitation focusing on continuity of care and attention to the cognitive as well as physiologic status results in effective and cost-effective rehabilitation. PMID- 11855272 TI - Severe brain injury rehabilitation. What's going to happen after critical care. AB - Treatment advances and technology will continue to decrease mortality in SBI in the future. A clearer line between lingering death and reasonable potential for recovery will only slowly reveal itself, and prediction will never be an exact science. Concerns about resource utilization and costs will continue to escalate. Critical care, acute care, and rehabilitation nurses will continue to live in this painful haze with the patients and their families. Nevertheless, critical care nurses can help by assisting families to understand the possible outcomes of SBI, the clinical state the patient is experiencing, how diagnosis and prognosis are related and how they are different, the indicators used to establish prognosis early on and then later in the course, and how prognosis is related to treatment decisions. Likewise, the critical care nurses can help the family begin to come to terms with the level of sophistication (or lack of precision, as the case may be) of prognostication, the agonizing time length factor involved, and the demanding prerequisites for level of consciousness assessment in the low functioning clinical states. PMID- 11855273 TI - Orthopedic trauma. Managing secondary medical problems. AB - Caring for the multiply injured orthopedic trauma patient is a challenge. Managing secondary medical problems associated with injury is enhanced when rehabilitation clinical nurse specialists, orthopedic advanced practice nurses, and the interdisciplinary team are consulted immediately following trauma. Educating the public on actions and behaviors that reduce the likelihood of accidents is an important role of the RN. Nurses play a key role in restoring the patient to the preinjury level of functioning or assisting the patient in adapting to temporary/permanent disability. Early intervention may result in shortened length of hospital stay, earlier return to work, restoration of family role function, and healthier psychologic and physical recovery. All RNs must be actively involved in educating the public on injury prevention, which includes teaching responsible behavior and the adverse effects of alcohol use. PMID- 11855274 TI - Catastrophic case management. A case study. AB - For the best clinical outcomes, catastrophic case management begins within the critical/intensive care units. The critical care nurse can facilitate this process by (1) documenting clearly, concisely, and legibly the patient's status in a holistic manner, as much of this information will be used to translate complex medical verbiage to clinical outcomes for financial authorization; (2) providing a receptive environment for the rehabilitation nurse case manager; (3) supporting the role of the rehabilitation nurse case manager in communications with the patient and family; (4) providing professional courtesy and dialogue when interacting with the rehabilitation nurse case manager; (5) understanding that although roles are different, the rehabilitation nurse case manager provides the continuous coordination of services and resources for optimal outcomes, and (6) including the rehabilitation nurse case manager in team or patient conferences so problems can be identified, allowing rehabilitation goals to be modified or expanded. Frequently, the rehabilitation nurse case manager is aware of the limitations of the health plans or capitated financial limitations of the benefit plans. With an informed understanding of the patient's problems, planning will maximize resources to achieve the best possible outcomes. For example, some benefits have a maximum number of home care visits annually or do not include acute rehabilitation hospitalization within the benefit plans. By matching the patient's needs to the benefit services within the time frame, resources are maximized. Most people are not aware of their health care plan services until the need arises. A rehabilitation nurse case manager possesses the expertise to advocate, negotiate, and use the benefits fittingly. PMID- 11855275 TI - Community re-entry after critical illness. AB - Community re-entry after a critical illness is a multifaceted ongoing process. Nurse case managers may have the opportunity for long-term follow-up. This is desirable because the individual with a disability must be considered as a developing and changing individual, physically, cognitively and spiritually. Coping and adapting to the many challenges is a continuous process as well. Although barriers persist in communities across the nation, much progress has been made in enabling persons with disabilities to live independently in the community. When value is placed on these individuals, they may live in the community as active and productive individuals who are seen for their abilities and not for their disabilities. Rehabilitation nurses have the opportunity to use many innovative approaches to access information, facilitate community resources and services, and ease transition for persons with disabilities who are returning to their communities. Nurses also have the opportunity to teach, empower, and advocate for individuals with disabilities so that they may achieve the highest level of independence possible. Successful community re-entry can only be achieved when the individual and his or her family are involved. Together with the rehabilitation team, they can return to the community as active and productive members of society or they may return simply because they belong to their community. Either way, they may enjoy quality of life as persons who have many abilities despite their disabilities. PMID- 11855276 TI - Managing the stress of caring for rehabilitation patients. AB - Dealing with stress is stressful! Evaluation is important to determine whether your plan for overpowering stress is working. Do you feel less stress as a result of what you have tried? Nurses use creativity every day in caring for their patients; applying that same spirit of creativity for stress reduction becomes personally meaningful. Stress is a universal condition, so friends and colleagues are good sources of ideas for stress management. Although stress is a human condition, unlikely to be totally eliminated, decreasing it as much as possible is important. Efforts in a stress reduction program can be met with great success. Recognizing the problem, assessing its magnitude, and then implementing selected strategies are all that is required. Overpowering stress in their lives is possible for nurses who are then healthier, happier, and even better caregivers. PMID- 11855277 TI - [Hysteresis of synaptic effectiveness--one of the possible mechanisms for associative learning?]. AB - The conditions on the occurrence of hysteresis of the amount of phosphorylated receptors were determined depending on the rhythmic frequency of presynaptic neuronal activity using the full and simplified models of synaptic plasticity for pyramidal hippocampal neurones. It is assumed that this phenomenon lies at the basis of the conservation of long-term potentiation. PMID- 11855278 TI - [Reception of images by the visual system in the imaginary plane in front of and behind the real plane]. AB - It was shown that our visual system and brain are able to initiate the appearance of images of pictures in an imaginary plane simultaneously behind and before their real plane and continuously watch their independent displacement. PMID- 11855279 TI - [Instability of a three-dimnsional linear vortex in a simple model of a heterogenous excitable medium]. AB - We studied the dynamics of three-dimensional linear vortex in a heterogeneous excitable medium. Using the Aliev-Panfilov model we showed that even a small heterogeneity in excitation threshold can lead to drastic deformations of the vortex filament and an instability in its behavior. PMID- 11855280 TI - [Molecular nature of the cryotolerant lake amphipod Gammarus lacustris]. AB - Gammarus lacustris were studied in an attempt to identify the mechanism of subzero temperature adaptation, which enables this species to survive in winter under natural conditions. G. lacustris hemolymph contained a high concentration of glucose (42.4 mg/ml of hemolymph). In addition, galactose, mannose, xylose, zellobiose, and unidentifiable disaccharides and oligosaccharides were found, though at much lower concentrations (0.94-3.26 mg/ml). The glycerol concentration in the gammarus hemolymph was about 5-20 times as low as in many other animals surviving at subzero temperatures (16.2 mg/ml). Other sugars and polyhydric alcohols were discovered only in traces amounts in the hemolymph. Antifreeze proteins were not found in the gammarus. PMID- 11855281 TI - [Effect of amino acids and low molecular weight peptides on the dynamics of water]. AB - The effect of amino acids and low-molecular-weight peptides on the dynamics of water was studied. Water medium together with molecules of dipeptides and amino acids dissolved in it is considered as a complex of interacting anharmonic oscillators. It was shown that the temperature behaviour of this system is determined by nonlinear resonances, which give rise to both the phenomenon of self-synchronization in the whole system or its part and to the phenomena of phase instability and coherence decay, depending on the store of oscillatory energy. Dissolved molecules are also involved in these oscillations, and if the frequencies and amplitudes of their oscillations are within the range in which nonlinear resonance occurs, they can affect the movement of the whole system. PMID- 11855282 TI - [Effect of weak and superweak magnetic fields on intensity and asexual reproduction of the planarian Dugesia tigrina]. AB - It was shown that the exposure to combined weak and extraweak magnetic fields (permanent component 42 microT; variable component of an amplitude of 100 nT, frequency 1-60 Hz) increases the intensity of asexual propagation of planarians Dugesia tigrina. The effect of combined magnetic fields is most pronounced at frequencies of 1, 3.7, and 32 Hz. The presence of concomitant technogeneous fields (50 Hz, 30 nT) does not markedly influence the effects of weak magnetic fields with a small variable component. Upon realization of effects of weak magnetic fields, their both components are of great importance; the absence of one (permanent) component changes the sing of the effect to the opposite. The transfer of the effect to planarians through water pretreated with magnetic fields probably indicates that aqueous medium is involved in the realization of biological effects of weak magnetic fields. PMID- 11855283 TI - [Generation of negative air ions by wheat seedlings in a high voltage electrization of soil]. AB - It was shown that plantlets of wheat (Triticum vulgare) are capable of generating negative aeroions during the electrization of soil by high-voltage impulses. Soil electrization was carried out either from the moment of planting of seeds or from the appearance of the first seedlings. The concentration of negative ions was measured in the air at a distance of 50 cm from plants. In both variants, similar growth-related changes in the concentration of negative ions were observed. The generation of negative ions began on day 6 after the planting of seeds and reached a concentration of 380 x 10(3) ion/cm3. During the next three days, this level remained unchanged. On day 10, the generation of negative aeroions increased abruptly; on days 10-14, it was twofold as high as on days 7-9. The level of generation of negative aeroions by plants stimulated from the moment of appearance of plantlets was 5-8% higher than by plants stimulated from the moment of planting. The intensity of generation of negative aeroions upon additional illumination and in full darkness remained unchanged. PMID- 11855284 TI - [Correlation of the width of annual tree rings with climatic changes and solar activity]. AB - Fluctuations in the width of annual tree rings in the time interval from the eighth century till the present time were analyzed on the basis of dendroclimatic scales. Periods of extremely high and low growth of trees were distinguished, and their correlations with heliogeophysical factors were studied. The major climatic parameters determining the growth of trees are warmth and moisture; a small gain is observed in cold and droughty periods. No correlations between the width of tree rings and the minima and maxima of solar activity cycles were observed with the method used. PMID- 11855285 TI - [Magnesium (Mg2+) cofactor in a triplet state reduces the proton of a coordinated water molecule to the hydrogen atom and pushes it out of the complex at a high speed]. AB - Quantum chemistry methods [RHF/UHF + MP4(FULL), DFT:B3LYP] with the 6-311+ +G**(p,d) basis set were used to elucidate the properties of six coordinated Mg2+ complexes with water, glutamic acid and ATP/GTP in singlet (S) and triplet (T) states. In the triplet state, the magnesium complex concentrates its spin density on a coordinated water molecule (inner or outer coordination shell). Within the molecule, a redox reaction occurs, and one of the hydrogen atoms is pushed out of the complex at a speed of approximately 125 m/s. In water solution, the energy of the triplet state is higher than that of the singlet state. In a mixed environment composed of water, amino acids and ATP/GTR, the energy of the magnesium complex in the triplet state is lower than that in the singlet state by 1.5-2.0 kcal/mol. A little difference in T and S states allows the Mg(2+)-ATP/GTP complex to switch easily between two reaction mechanisms. PMID- 11855286 TI - [Chemiluminescence in oxidation of luminol by chloramine derivatives of biogenic compounds]. AB - Chloramine derivatives of amino acids induce chemiluminescence of a luminol solution. The chemiluminescence is more prolonged than the emission of luminol produced by hypochlorite. Persistent chemiluminescence also appears under the action of hypochlorite on a mixture of luminol and amino acids. It is assumed that the chemiluminescence of luminol in suspensions of stimulated phagocytes may be associated with its oxidation by chloramines. PMID- 11855287 TI - [A model of the dynamics of intracellular calcium based on refined kinetics of inhibiting the inositol-1,4,5-trisphoshate-sensitive calcium channel]. AB - Several new models of intracellular calcium dynamics based on refined inositol 1,4,5-triphosphate-sensitive calcium channel kinetics were studied. The refined kinetic schemes take into account that a cytosolic calcium cannot inhibit inositol-1,4,5-triphosphate receptors when they are bound to inositol-1,4,5 triphosphate. The mathematical analysis of intracellular calcium dynamics based on one of these schemes allowed us to show how different types of Ca response to extracellular stimuli, such as excitability, oscillations, sustained elevation of Ca and frequency encoding can arise with a reasonably good fit to experimental data. PMID- 11855288 TI - [Features of organic nonelectrolyte binding to the erythrocyte membrane]. AB - The capacity of erythrocyte membranes for organic nonelectrolytes from different chemical groups of chemical compounds was studied by the spin probe method and scanning electron microscopy. Hydrophobic spin-labeled derivatives of gamma carbolin and stearic acid and the screened phenol antioxidant fenozan-1 were used as nonelectrolytes. Based on the analysis of electron spin resonance spectra of the hydrophobic spin-labeled nonelectrolytes and electron micrographs of erythrocytes, differences in the capacity of distribution regions in the intramembrane space of the derivative of gamma-carbolin and fenozan-1, on the one hand, and the spin-labeled derivative of fatty acid, on the other hand, were found. The first group has at least two membrane distribution regions, whereas in the second case only one type of distribution was found. The influence of limited membrane capacity on the realization of biological activity of organic nonelectrolytes is discussed. PMID- 11855289 TI - [Change in the dynamics of a spin probe in complement-dependent lysis of a liposome]. AB - The mobility of phospholipid chains in membranes of liposomes consisting of egg lecitin, cholesterol, dicetylphosphate, sensitized by the lipopolysaccharide antigen F. tularensis by the action of a homologous antiserum and a rabbit complement preparation was studied using 5- and 16-doxylstearate spin probes. It was shown that, during the immune lysis of liposome membranes, changes in the dynamics of spin probes occur, which correlate with the formation of transmembrane channels and exit of the fluorescent marker from the interior of liposomes. It was found that the ratio of the intensities I1/I2 of two low-field extrema in the ESR spectrum is most sensitive to changes in the liposome membrane that are induced by immune components. PMID- 11855291 TI - [Microcalorimetric study of native cells of the microalgae Spirulina platensis in the temperature range (2-55)C]. AB - Thermal effects occurring upon heating a culture of blue-green microalgae Spirulina platensis in the temperature range 5-55 degrees C were studied. Under these conditions, an intensive heat evolution was observed. The heat evolution versus-temperature curve has a peak with a maximum at approximately 45 degrees C and two distinct shoulders at approximately 25 and 40 degrees C. It was found that heat evolution Q at heating rates below 0.083 degree C/min does not change and is (381 +/- 20) J/g of dry biomass. It was concluded that this value is optimal for maintaining the cell viability (in particular, respiration) under anaerobic conditions, in the dark and in the stationary regime. PMID- 11855290 TI - [Enhancement of luminol-dependent chemiluminescence in an aqueous NaCl/H2O2 solution by argon]. AB - It was found that the bubbling of argon through NaCl/H2O2 aqueous solutions results in the development of intense sustained luminol-dependent chemiluminescence. Bubbling of nitrogen and air through similar solutions does not result in such effect. The relationship between argon-supported chemiluminescence and initial concentrations of NaCl and H2O2 is characterized by threshold effects. In NaCl/H2O2 solutions blown with argon, hypochlorite was found, indicating that argon intensifies the reaction of chloride oxidation with H2O2. It is suggested that the enhancement of this reaction in aqueous solutions saturated with argon is related to specific changes in the properties of water, which is a highly nonequilibrium system. Possible consequences of relatively high concentrations of argon in the atmosphere for the chemical processes that occur in aqueous systems and, in particular, living systems are discussed. PMID- 11855292 TI - [Carbon isotope (13C/12C) effect of photorespiration in photosynthetic organisms. Evidence for existence, probable mechanism]. AB - Experimental evidence in favor of the new phenomenon predicted for photosynthesizing organisms, the fractionation of carbon isotopes in photorespiration is presented. A possible mechanism of this process is discussed. The fractionation of carbon in isotopes photorespiration occurs in the oxygenase phase of the functioning of ribulosebisphosphate carboxylase/oxygenase (rubisco), the key enzyme of photosynthesis, which is capable to act as carboxylase and oxygenase. Which function of the enzyme is active depends on CO2/O2 concentration ratio, which periodically changes in a cell. The key reaction in the mechanism of carbon isotope fractionation in photorespiration is glycine decarboxylation, which results in the splitting and removal from the cell of CO2 enriched with 12C and the accumulation of 13C photorespiratory carbon flow. The coupling of photorespiration and CO2 photoassimilation gives rise to two isotopically different carbon flows, which fill up separate carbohydrate pools, which are the sources of carbon in the following syntheses in the dark phase of photosynthesis. This enables one to identify, from the carbon isotope ratio of metabolites, their involvement in the photorespiratory and assimilatory carbon flows, to investigate the pathways of carbon metabolism, and to estimate more thoroughly the biosynthetic role of photorespiration. PMID- 11855293 TI - [Suppression of nonspecific resistance of the body under the effect of extremely high frequency electromagnetic radiation of low intensity]. AB - The dynamics of leukocyte number and functional activity of peripheral blood neutrophils under whole-body exposure of healthy mice to low-intensity extremely high-frequency electromagnetic radiation (EHF EMR, 42.0 GHz, 0.15 mW/cm2, 20 min daily) was studied. It was shown that the phagocytic activity of peripheral blood neutrophils was suppressed by about 50% (p < 0.01 as compared with the sham exposed control) in 2-3 h after the single exposure to EHF EMR. The effect persisted for 1 day after the exposure, and then the phagocytic activity of neutrophils returned to the norm within 3 days. A significant modification of the leukocyte blood profile in mice exposed to EHF EMR for 5 days was observed after the cessation of exposures: the number of leukocytes increased by 44% (p < 0.05 as compared with sham-exposed animals), mostly due to an increase in the lymphocyte content. The supposition was made that EHF EMR effects can be mediated via the metabolic systems of arachidonic acid and the stimulation of adenylate cyclase activity, with subsequent increase in the intracellular cAMP level. The results indicated that the whole-body exposure of healthy mice to low-intensity EHF EMR has a profound effect on the indices of nonspecific immunity. PMID- 11855294 TI - [Effect of electromagnetic waves in the centimeter range on the production of tumor necrosis factor and interleukin-3 in immunized mice]. AB - The effect of prolonged treatment with weak microwaves on the production of tumor necrosis factor and interleukin-3 in peritoneal macrophages and T cells of male NMRI mice twice immunized by affinity-purified carboanhydrase was studied. Against the back ground of a high titer of antibody production, a significant increase in the production of tumor necrosis factor in peritoneal macrophages and splenic T lymphocytes of immunized mice was revealed, and a much stronger effect was observed for irradiated immunized animals. A tendency to increased secretion of interleukin-3 for unirradiated and irradiated immunized animals was found; in the latter group of animals, the effect being more pronounced. The stimulation of production of the cytokins, especially tumor necrosis factor, by combination of antigenic stimulation and microwaves can be used in adjuvant therapy of various immune diseases. PMID- 11855295 TI - [Flow microchamber for the Linnic interference microscope]. AB - A flow microchamber for the Linnic microscope was constructed. The volume of the microchamber is 6-10 microliters. The microchamber may be used in two working regimes: (a) under conditions of periodical change of solutions during the analysis of the same object and (b) under conditions of continuous flow of solutions. The microchamber was used for studying the birefringence of skeletal muscle sarcomere at rigor and relaxed states. PMID- 11855297 TI - [Superweak irradiation of marine invertebrates]. AB - The inhibitory analysis of the spontaneous ("ultraweak") and the luminole-induced chemiluminescence of marine Sycon sponges and Aiptasia actinias supports the idea that ultraweak photon emission of marine invertebrates is a consequence of Ca(2+) dependent processes related to the interaction of reactive oxygen species with some endogenous fluorophore substrates. PMID- 11855296 TI - [Low-frequency oscillations of the energy-conserving reactions of photosynthesis and level of indoles in wheat ontogenesis]. AB - Changes in the rates of photophosphorylation and the content of indoles in wheat ontogeny were studied. On the basis of experimental data, a phenomenological parametrical model was elaborated. It was found that the temporal changes in the parameters examined are nonmonotonous and include an oscillatory component. The amplitudes and frequencies of the oscillatory components and the parameters of the model were estimated. PMID- 11855298 TI - [A computer method for assessing the average rate of movement of Paramecium caudatum by a cross-sectional light beam]. AB - The possibility of using the motility of Paramecium caudatum for computer biotesting of toxic substances was studied. The method is based on measuring the number and duration of impulses that arise when protozoa chaotically moving in a layer 0.5 mm thick cross a red light beam (1 mm in diameter). It was found that, in the presence of Cu2+ ions and menadione bisulfite, a compound having prooxidant properties, the number of impulses decreased with a concurrent increase in the duration of impulses. The high correlation between the number of impulses and the reciprocal of impulse duration indicates that the motility of paramecium by the action of Cu2+ ions and menadione slow down, and even an immobilization of some of them occurs. The mean swimming velocity of protozoa was reduced by half within 20 min by the action of Cu2+ at a concentration of 90 microM (6 mg/l of Cu2+ ions) and within 80 min by the action of menadione bisulfite at a concentration of 50 microM. It was shown that the slowing down of motility, which indicates the presence of a toxicant in the medium, is a faster physiological reaction than cell death. PMID- 11855299 TI - [Workers' health care and prophylaxis of occupational diseases at the present time]. AB - Nowadays industrial medicine faces significant and novel goals concerning workers' health care and occupational diseases prophylaxis. These goals result from the corresponding Federal Laws and could be solved according to justified prognosis for research development up to 2001. PMID- 11855300 TI - [On a prognostic value of some phenotype markers for blood groups, neutrophils reactivity and lipid peroxidation in pneumoconiosis]. AB - The article represents results of study determining biologic markers through examining phenotypic features of blood groups, blood neutrophils biocidity, lipid peroxidation and antioxidant defense in RBC. Those features result in high sensitivity or resistance to dust, in probability of early or late pneumoconiosis development. PMID- 11855301 TI - [Role of hormonal factors in pulmonary angiopathy development during anthrax silicosis]. AB - The article represents results of radionuclide study of hormonal levels and pulmonary microcirculation in anthracosilicosis. The authors discuss mechanisms underlying influence of vasoactive hormones on pulmonary microcirculation. The authors revealed relationships between hormonal and vascular disorders in anthracosilicosis. PMID- 11855302 TI - [Influence of occupational dusts load on state of antioxidant system in miners engaged in diamond extraction industry]. AB - Miners of diamond extraction industry in Yakutia demonstrated higher intensity of lipid peroxidation. Longstanding compensatory adaptive response to increased free radical oxidation and to unfavorable climate factors is higher superoxide dismutase and catalase and changes in protein metabolism. Lack of C and E vitamins in miners results from increased utilization due to strain of antioxidant defence mechanisms. PMID- 11855303 TI - [Problem of individual predisposition to occupational chronic bronchitis (review of literature)]. PMID- 11855304 TI - [On prevalence of pneumoconiosis in cement plants (review of literature)]. PMID- 11855305 TI - [Epidemiologic approaches to diagnosis of diseases dependent on ecologic factors]. AB - The article deals with peculiarities in diagnosis of diseases caused by environmental hazards. The diagnosis covers population level and includes evaluation of environmental quality. Analysis of health for whole population and for risk groups enables to reveal "indicator" diseases. Social and hygienic monitoring, complex sanitary and ecologic examination recommend disclosure of facts and causes for any "nonspecific" diseases, definition of disorders caused by specific environmental hazards. The authors consider unity of hygienic and epidemiologic analysis for causative relationships in "human-environment" system in evaluation of noninfectious risk factors. PMID- 11855306 TI - [Hygienic evaluation of technologic laser devices]. AB - The author necessitates participation of occupational hygienists early in design of industrial laser equipment, laser devices, in order to create safe technologic equipment. PMID- 11855307 TI - [Studies of decomposition kinetics and photometric detection of chlorsulphanylisocyanate in air]. PMID- 11855308 TI - [On carcinogenic effects of sulfate copper and nickel ore]. AB - Underground mines are places for extraction of three types of copper and nickel sulfide ores: rich, copper and incorporated--those three ones differ in mimeralogic and chemical contents. Experimental evaluation of carcinogenic effects obtained during intratracheal administration of copper and incorporated ores indicates the certain carcinogenic activity of the dust with 0.49-1.14% nickel and 1.9-5.8% copper content--therefore a unified MAC for all the stated dust types could be suggested. PMID- 11855309 TI - [The role of cytokines in pathogenesis of nonspecific ulcerative colitis]. AB - Etiology of nonspecific ulcerous colitis (NSUC) as well as its pathogenetic mechanisms remain obscure. Cytokines are presently shown to play an essential role in development of inflammation. Cytokines are produced by the cells in the inflammatory infiltrate in NSUC, therefore, changes of their level in the blood and biopsies from colon mucosa is naturally determined. Current investigations register various changes in the levels of both proinflammatory and antiinflammatory cytokines. Further studies of unknown mechanisms of cytokines action may contribute to design of new methods of NSUC management. PMID- 11855310 TI - [Hemodynamics of the minor circulation in patients with gastroduodenal ulcer on hypoxic therapy]. AB - Adaptation of patients with gastroduodenal ulcer (GDU) to intermittent normobaric hypoxia is effective in the treatment of this disease. Some GDU patients have functional pulmonary hypertension. Single procedure and a course of normobaric hypoxic therapy have no negative effect on pulmonary hemodynamics. Overloading of the right heart in GDU patients does not contraindicate a course of breathing gas hypoxic mixture created by Strelkov's apparatus. PMID- 11855311 TI - [Genotypes of ACE gene in Kazakh and Uygur patients with ischemic heart disease]. AB - Frequency distribution of angiotensin-converting enzyme (ACE) gene genotypes and alleles was studied in patients with ischemic heart disease (IHD) and healthy subjects of Kazakh and Yugur nationalities. Polymorphism of ACE gene was determined by means of polymerase chain reaction. 88 IHD and 72 healthy Kazakhs and 70 and 70 Yugurs, respectively, were examined. Incidence of DD genotype in the studied groups was the least. This agrees with the literature data about low occurrence of genotype DD in Asian nationalities. Frequency of ACE gene genotypes in IHD and healthy subjects of Kazakh and Yugur nationalities do not differ significantly. Thus, polymorphism of ACE gene is unrelated to IHD in the examinees. PMID- 11855312 TI - [Cardiac pathology in various in various pathogenetic subtypes of ischemic stroke]. AB - Heart pathology was studied in various pathogenetic subtypes of ischemic stroke. A total of 330 stroke patients were divided into 4 groups. Group 1 consisted of 104 (31.5%) patients with cardioembolic stroke, group 2--of 72 (21.8%) with hemodynamic, group 3--of 71 (21.5%) patients with atherothrombotic and group 4- of 83 (25.2%) patients with lacunar stroke. Cardiac pathology was found in all patients of group 1 (ischemic heart disease-50.0%, inflammatory-infectious lesions-27.9%, other lesions-22.1%), in 49 (68.1%) patients of group 2, 36 (50.7%) of group 3 and 31 (37.3%) of group 4. Ischemic heart disease was prevalent (85.7, 86.1, 83.9% vs group 1). Potential sources of cardiogenic cerebral embolia were found in patients of all the groups: 100, 62.5, 43.7 and 33.7%, respectively. Thus, most patients with ischemic stroke have different heart defects related to the subtype of ischemic stroke. Potential causes of cardiocerebral embolism can be detected in any subtype of ischemic stroke. PMID- 11855313 TI - [Effectiveness and safety of amlodipine in secondary prevention in patients with myocardial infarction]. AB - Amlodipin (norvask) effectiveness was studied in 18 patients who survived primary Q-myocardial infarction. The 2-year treatment produced a positive effect in 66.6% patients, a negative one--in 11.1%. One patient died, a repeat non-fatal MI developed in 2 patients. Complications were registered in 1 patient. The drug reduced the number of angina attacks, amount of nitroglycerin (by 46.6 and 45.5%, respectively), the tolerance increased by 26.7, cardiac performance by 59.9%. It also led to a fall of both systolic and diastolic blood pressure (by 14.0 and 9.9%, respectively). Heart rate remained unchanged. Left ventricle underwent adaptive remodeling, myocardial ischemia diminished in the number of episodes and their duration. Frequent atrial and rare ventricular extrasystole disappeared in 1 patient. PMID- 11855314 TI - [Clinical implication of the body mass and waist/thigh indices in hypertensive patients: correlations with blood uric acid levels]. AB - Body mass index (BMI) and waist/thigh circumference (WTC) were estimated in 104 patients with mild and moderate arterial hypertension (AH) lasting for 8.4 +/- 3.2 years (63 men, 41 women, mean age 50.2(+)-9.6 years). Blood concentration of uric acid was tested with enzymes (Boehringer Mannheim kits). Only 5 of 104 examinees had BMI under 25. The majority of the patients (79%) had high WTC. Hyperuricemia was detected in 32 of 104 patients. Uric acid level in the blood correlates (r = 0.43) with WTC but not with BMI. PMID- 11855315 TI - [Thrombocytic lipids in different clinical manifestations of myocardial infarction]. AB - The stady was made of changes in lipid composition of platelet membranes in various clinical manifestations of myocardial infarction (MI). Phospholipids, cholesterol and its ethers in platelet membranes were studied which thinlayer chromatography in 98 MI patients aged 30-60 years and 20 healthy subjects aged 32 56 years. In acute MI platelet membrane shows metabolic disturbances while subacute phase and scarring are characterized by positive membrane processes. In lethal MI membrane disembolism is irreversible. Thus, membrano-destructive alterations in platelets correlate with MI clinical symptoms and outcome. PMID- 11855316 TI - [The status of respiratory organs in hypertensive patients]. AB - The condition of the respiratory system in essential hypertension was studied with the tests of general hemodynamics, echocardiography, zonal rheography of the lungs and rheospirography with estimation of ventilation-perfusion relations. Hypertension was found to affect not only cardiovascular but also respiratory system. Hypertension-related respiratory disorders present as vascular dysfunction in the lungs, restrictive defects of pulmonary ventilation, imbalance between the ventilatory reserve and reserve of the pulmonary and systemic blood flow with progression of arterial hypertension. PMID- 11855318 TI - [Exogenic toxic alveolitis: clinical symptoms and diagnosis]. AB - X-ray examination of 36 patients with exogenic toxic alveolitis (ETA) has proved that ETA may run both in x-ray positive and x-ray negative forms which run with similar clinical manifestations. It is important in ETA diagnosis to measure pressure in the pulmonary artery and to examine pulmonary perfusion. In complicated cases it is recommended to conduct histological verification of the diagnosis in bronchial or open lung biopsy. PMID- 11855317 TI - [Cardiovascular and metabolic effects in nebulizer therapy of bronchial asthma]. AB - The aim of the study was examination of cardiovascular effects of nebulizer salbutamol (ventolin) therapy of bronchial asthma (BA). The efficiency of the drug was assessed in 38 BA patients by changes in clinical symptoms and rise of peak volume expiration rate. Before and after ventolin treatment (2.5-5 mg), measurements were made of central hemodynamics, microhemodynamics, cardiac arrhythmia, plasma electrolytes. The results of the tests indicate that nebulizer treatment with ventolin effectively and safely eliminates bronchoobstructive syndrome in BA patients. Systolic and diastolic arterial pressure, left ventricular end diastolic pressure, ventricular extrasystole declined, supraventricular extrasystole rose. Microcirculatory effects were both positive and negative. Plasma concentration of potassium ions reduced insignificantly. PMID- 11855319 TI - [Local skin blood flow in hypokinesis]. AB - Functional indices of peripheral autonomic nervous system responsible for regulation of tissue circulation were studied with doppler laser-flowmetry in patients exposed to long-term hypokinesia. Valsalva test, orthostatic test, cold test, etc., have discovered shifts in the system regulating local skin blood flow. Compared to control, changes in the blood flow were considerable and its recovery went very slowly. PMID- 11855321 TI - [Pathogenesis of HBV- and HCV-infections in alcoholic disease of the liver]. PMID- 11855322 TI - [Endothelial dysfunction in pathogenesis of duodenal ulcer]. AB - It is shown that in patients with ulcer associated with Helicobacter pylori (HP) there is a close correlation between the severity of the lesion of gastroduodenal protective mucous barrier and that of endothelial dysfunction manifesting in elevated level of endothelin-1, serum levels of TBK-active products, inhibition of blood flow and narrowing of the celiac trunk. The correlation becomes stronger with expanding contamination of gastroduodenal mucosa with HP. Thus, HP may participate in breaking the protective mucous barrier in endothelial dysfunction. PMID- 11855320 TI - [Piracetam in combined pathogenetic therapy of recurrent duodenal ulcer]. AB - Duodenal ulcer cure, as a systemic gastroenterologic disease, can be achieved in some patients by the addition of the nootropic drug piracetam to current antisecretory and antihelicobacter therapy. Piracetam corrects vegetative and psychoemotional disorders in duodenal ulcer, normalizes gastric motility, has an antioxidant effect and improves cerebral circulation. An optimal effect on clinico-endoscopic manifestations of recurrent duodenal ulcer was achieved in combination of piracetam with current antisecretory (omeprazole) and antihelicobacter (de-nol, amoxicillin, metronidazole) medicines. Such combination improves both short- and long-term outcomes of duodenal ulcer treatment. PMID- 11855323 TI - [Administration of various dosages of retard isosorbide-5-mononitrate in patients with stable angina pectoris on combined therapy]. AB - We studied efficiency of a retard form of isosorbide-5-mononitrate (Mono Mac 50D) in patients with stable angina pectoris (NYHA class II and III). This efficiency was assessed by clinical examinations and 24-h Holter ECG monitoring before and after the treatment. 16 patients with angina of functional class II and 12 patients with functional class III received combined treatment: prolonged mononitrate (Mono Mac 50D), disaggregants, beta-blockers, ACE inhibitors and calcium antagonists (on demand). In angina functional class II Mono Mac 50D was given in a single dose 50 mg/day (1 tablet) in the morning for 4 weeks, in functional class III angina the drug was taken for 2 weeks in a dose 50 mg/day, the next two weeks in a dose 100 mg/day once in the morning. The other drugs were taken in moderate recommended doses. After two weeks of treatment with 50 mg/day Mono Mac 50D angina symptoms attenuated (in functional class II) and the patients' condition improved (in functional class III). In elevation of the dose to 100 mg/day the latter patients improved still greater. We think it valid to use a single 50 mg/day dose in angina functional class II and 100 mg once a day in functional class III. PMID- 11855324 TI - [50 years after ante-thoracic esophagoplasty]. AB - Fifty years after antethoracic esophagoplasty the patient's quality of life was rather satisfactory. PMID- 11855325 TI - [On the manifestation of diabetes mellitus in a patient with severe course of acute Flexner's dysentery]. AB - Diabetes mellitus (DM) was diagnosed in a 54-year-old man with acute Flexner's dysentery (AFD). A severe course of AFD impedes early clinical diagnosis of DM detected only upon appearance of life-threatening ketoacidosis. PMID- 11855326 TI - [A case of primary pulmonary hypertension]. AB - Primary lung hypertension (PLH) is a rare disease of unknown etiology seen largely in young persons. PLH shows rapid progression with lethal outcome 3-5 years after registration of the first symptoms. Early diagnosis of the disease is difficult. A case described in the article illustrates real difficulties in PLH diagnosis and treatment. The patient several times lost consciousness for a short time at insignificant exercise. This phenomenon has not been described in the literature so far. PMID- 11855328 TI - [Problems of congestive heart failure at the XXII Congress of European Cardiologic Society (Amsterdam, Netherlands, August 2000)]. PMID- 11855327 TI - [A severe amebic abscess of the left lobe of the liver]. PMID- 11855329 TI - [Minor heart defects]. AB - Minor anomalies of the heart are morphological changes of the structure and cause functional cardiovascular disorders. Polyorganic disorders and phenotype are indicated typical for congenital defects of the connective tissue. The opinions on mechanisms of development of pathological symptom complexes in minor anomalies of the heart vary. Further studies are necessary. PMID- 11855330 TI - [The change in the activity of angiotensin converting enzyme in patients with pneumonia and chronic obstructive pulmonary diseases]. AB - The activity of angiotensin-converting enzyme (ACE) was measured in the serum and bronchial contents of 69 patients with pneumonia and 77 with chronic obstructive pulmonary diseases (COPD). ACE activity was decreased both in the blood and bronchial contents during the acute phase of pneumonia. With resolution of the inflammatory process, ACE activity normalized. In patients with COPD, the activity of ACE is decreased during remission in comparison with the mean values in the population. During COPD exacerbation the activity of ACE increases both in the blood and bronchial contents. Changes in ACE activity in pneumonia and COPD are more pronounced in the bronchial contents than in the blood. Presumably alteration of the enzyme concentration is the only cause of alteration of its activity in patients with COPD and pneumonia. PMID- 11855331 TI - [Characteristics of salivary protein spectrum in patients with depressive disorders (in search for biological markers of depression)]. AB - Salivary protein spectrum was studied by polyacrylamide gel electrophoresis in 80 patients with psychogenic affective disorders, 80 with rheumatic and 10 with reactive arthritis, and 9 patients with chronic renal insufficiency in the presence of depression. Control group consisted of 60 healthy subjects. The results indicate that depression is associated with decreased content of protein in different fractions of the spectrum, the most marked decrease being observed for the fraction with molecular weight of about 55 kD. The optical density of this fraction decreases to 0.11 units in depression (versus 0.17 +/- 0.02 in the control), which suggests that this method can be used for the diagnosis of depressions. PMID- 11855332 TI - [Analysis of fatty acid composition of sweat lipids in children and adults with skin disease]. AB - Gas-chromatographic analysis of fatty acid composition of sweat lipids in children and adults with neurodermatitis, atopic dermatitis, and eczema showed that sweat can be used as a new noninvasive biological object for evaluation of lipid metabolism disorders. PMID- 11855333 TI - [Blood level of lipid peroxidation products as an indicator of resistance to infectious inflammatory complications of transurethral electroresection of the prostate]. AB - Study of relationship between the content of lipid peroxidation (LPO) products in the blood of patients with benign prostatic hyperplasia (BPH) and resistance to infectious inflammatory complications (IIC) of transurethral electroresection of the prostate showed that decreased content of circulating lipoperoxides promoted the development of postoperative IIC. Before the operation blood levels of LPO products were increased in BPH patients who did not develop IIC postoperation in comparison with normal controls. Three intravenous injections of ascorbic acid in a single dose of 1000 mg after transurethral electroresection of the prostate led to an increase in the blood level of LPO products and promoted a decrease in the incidence of postoperative IIC. PMID- 11855334 TI - [Diagnostic significance of analysis the energy metabolism substrates in biological fluids]. AB - Changes in energy metabolism substrates in biological fluids (venous and umbilical blood, urine, and amniotic fluid) were studied. Lactate concentrations were 2-4--fold increased in venous blood and 2-fold in umbilical blood. Urinary excretion of energy metabolism substrata was increased in women with gestosis. An increase of lactate concentration above 0.110 mmol/liter and of isocitrate concentration above 0.238 mmol/liter indicated fetal hypoxia. PMID- 11855335 TI - [Use of computer image analysis for differential cytological diagnosis and prognosis of hepatocellular carcinoma (review of literature)]. PMID- 11855336 TI - [HIV drug resistance and laboratory methods for its detection (Lecture)]. PMID- 11855337 TI - [Diagnostic significance of albumin transporting characteristics and blood level of troponin T in myocardial infarction]. AB - Serum albumin binding capacity (ABC) was studied using K-35 hydrophobic fluorescent probe in order evaluate the passive transfer and absorption of saturated fatty acids (s-FA) by the cells. ABC decreases on day 3 of myocardial infarction and remains low till discharge and is paralleled by hyperTG. We believe that ABC decrease reflects activation of passive transfer and absorption of s-FA by the cells during simultaneous blocking (presumably by acute phase proteins) of active apoE/B-100 receptor endocytosis of s-FA in the form of TG as VLDL remnants. Decrease of ABC reflects a later than erythrocyte sedimentation rate and acute phase proteins normalization of metabolic consequences of inflammation. ABC helps indirectly evaluate the blocking of active (receptor) absorption of s-FA in the form of TG and activation of passive absorption of s-FA from associations with albumin by the cells. PMID- 11855338 TI - [The potential of using "Cellprint" system for analysis of synovial fluid cell composition]. PMID- 11855339 TI - [Cytomorphological characteristics of ovarian dysgerminoma]. AB - Cytomorphological signs of ovarian dysherminoma are distinguished on suboperation material from 7 patients. Scrapings from removed ovarian tumors were stained after N. G. Alexeev and Pappenheim. Dysherminoma cytograms were characterized mainly by a combination of three cell types: large sharply polymorphic epithelioid with multiple clearly-seen nuclei, connective tissue, and lymphocyte like cells. PMID- 11855340 TI - [Study of proliferative activity and nucleolar organizers in the epidermis of patients with psoriasis]. AB - Time course of proliferative activity of the epidermis was studied in patients with psoriasis by analysis of histoautoradiography data and evaluation of the activity of the ribosome nucleolar organizers. The findings indicate that the proliferative activity is increased in psoriasis. The activity of the ribosome nucleolar organizers increases during the progressive stage of psoriasis and notably decreases during the stationary stage, while during regressive stage this activity is the same as in apparently normal (after resolution of eruptions) skin of psoriasis patients. The results indicate that evaluation of the activity of nucleolar organizers in psoriatic elements can serve as an important additional test in laboratory diagnosis of psoriasis stages. PMID- 11855341 TI - [Computed image analysis of neutrophils: alkaline phosphatase]. AB - Alkaline phosphatase (AP) activity was detected under optic microscope in neutrophilic leukocyte granules and intergranular cytoplasm. Computer analysis of the image showed that activation of neutrophils in the blood of patients with purulent peritonitis was paralleled by a drastic increase in the activity of AP and its intracellular redistribution. AP granules increased in size and their capacity to conglomeration increased. The enzyme activity increased both in the granules and diffuse zone of the cytoplasm. Total area of AP granules directly correlated with the size of the zone of the enzyme diffuse-granular location. PMID- 11855342 TI - [Nucleic acid content in peripheral blood lymphocytes from patients with infectious mononucleosis]. AB - An increase in RNA content in peripheral blood lymphocytes and increased number of macronuclear weakly basophilic cells (presumably due to increased content of B lymphocytes which, in contrast to sharply basophilic T-lymphocytes, are characterized by light basophilic cytoplasm) are observed in patients with infectious mononucleosis. Macronuclear forms represented equally by hyper- and hypobasophilic elements predominate the structure of atypical mononuclears. Increased content of DNA in the peripheral blood lymphocytes is due to increased number of cells with high DNA content in the patient's blood and appearance of cells with 52-65 arb. units DNA (blast-transformed atypical mononuclears infected with the virus) which are normally absent. PMID- 11855343 TI - [Standardization of immunophenotyping techniques for human blood and bone marrow cells. Part III. Immunocytochemical methods. Part IV. Normal value]. PMID- 11855344 TI - [A new method for total complement activity analysis and its clinical significance]. AB - A test system consisting of a short-term suspension culture of mobile cells (bovine semen) is suggested for evaluating the complement activity. A simple and effective rapid method for evaluating the complement activity is based on the use of this system. Clinical application of this method helps evaluate the disease severity and predict the patient's condition in various diseases. PMID- 11855345 TI - [Incidence of HIV infection in donors in 1996-2000]. PMID- 11855346 TI - [Protein markers of diabetic retinopathy]. AB - Total and 12 individual proteins were measured in the lacrimal fluid of children and adolescents with insulin-dependent diabetes mellitus. Lacrimal excretion of proteins helps diagnose the preclinical, nonproliferative, and preproliferative stages of diabetic retinopathy and predict the course of diabetic cataract. PMID- 11855347 TI - Role of abortion in fertility control. PMID- 11855348 TI - Rapid detection of Haemophilus influenzae type b in Bangladeshi children with pneumonia and meningitis by PCR and analysis of antimicrobial resistance. AB - A polymerase chain reaction (PCR) assay with primers from 'bexA' gene was compared with culture for the detection of Haemophilus influenzae type b (Hib) in clinical samples from children with pneumonia and meningitis. Of 200 sera (180 from pneumonia, 20 from non-pneumonia patients) tested by PCR (serum-PCR), Hib was detected in 15 of 16 blood culture-positive and in 6 blood culture-negative pneumonia cases. When compared with the results of blood culture, serum-PCR had sensitivity, specificity, and accuracy index of 93.7%, 96.7%, and 96.5% respectively. Of 120 cerebrospinal fluid (CSF) samples from meningitis patients tested by culture and PCR (CSF-PCR), the latter method could detect Hib in all 15 culture-positive and in 8 of 105 culture-negative cases, showing sensitivity, specificity, and accuracy index of 100%, 92.4%, and 94.4% respectively. The PCR result was available within a day. Antimicrobial susceptibility of Hib was determined by the disc-diffusion method. High rate of resistance to ampicillin (54.8%), chloramphenicol (48.4%), and co-trimoxazole (80.6%) was observed among 31 invasive Hib isolates with resistance to all 3 drugs (multiresistance) in 48.4% of the isolates. All the Hib isolates were susceptible to ceftriaxone. The study has shown that PCR is a rapid, sensitive and specific diagnostic test for Hib from clinical samples, and a combination of culture and PCR is necessary for the detection of Hib infections to the maximum extent for case management to reduce morbidity, mortality, and complications of the invasive Hib infections. A high prevalence of multiresistant Hib strains is a matter of concern. PMID- 11855349 TI - Malnutrition among pre-school children in Alexandria, Egypt. AB - The study was conducted in Alexandria, Egypt, to assess the current status of malnutrition among 1,217 pre-school children aged 6-71 months. A two-stage cluster-sampling technique was used for selecting the sample. Data on sociodemographic and environmental characteristics of the family, morbidity profiles, and breast-feeding patterns were collected from mothers of the children. Anthropometric measurements were performed, and the prevalence of malnutrition was assessed using three indicators, such as stunting, under-weight, and wasting, following the WHO guidelines and cut-off points. Simple and multiple regression analyses were done for examining the factors associated with the occurrence of malnutrition using principal component factor analysis with varimax rotation. Stunting, under-weight, and wasting were observed in 15%, 7.3%, and 3.6% of the children respectively. High-socioeconomic condition was associated with low prevalence of stunting and underweight (OR = 0.67, confidence interval (CI) = 0.55-0.8 and OR = 0.75, CI = 0.58-0.96 respectively). Good environmental condition was associated with a lower stunting rate (OR = 0.83, CI = 0.72-0.96). Increased age of child and living in a non-squatter area were associated with wasting (OR = 1.02, CI = 1.001-1.03 and OR = 0.38, CI = 0.15-0.97 respectively). Interventions to improve socioeconomic and environmental situations are recommended to reduce the already low level of protein-energy malnutrition further. PMID- 11855350 TI - Ordeal of women for induced abortion in a rural area of Bangladesh. AB - The study was carried out to document the context of induced abortion, nature of its management, and post-abortion complications in Matlab, Bangladesh. The study included all 91 cases of induced abortion that took place in the study area from July to October 1995. Information was collected from women within 60 days after the abortion. A physician carried out in-depth interviews and physical examinations of 20 randomly-selected cases. The findings depicted a complex context, within which the women had to go for an abortion. In most cases, the complete lack of use or lack of use-effectiveness of family-planning methods resulted in unwanted pregnancies. The women in desperation sought abortion services from traditional sources first. When their conditions worsened, they contacted the available modern service facilities. At times, it was too late and led to serious health consequences. Limited access to safe abortion services, together with an absence of social support, put women in a life-threatening situation. Prevention of unwanted pregnancies and access to safe abortion services are needed to improve the situation. PMID- 11855351 TI - Socioeconomic and health implications of adult deaths in families of rural Bangladesh. AB - Effects of adult deaths on subsequent health and socioeconomic well-being of rural families of Bangladesh were examined. Data for this study were drawn from the longitudinal Sample Registration System (SRS) operational in two rural areas of the then MCH-FP Extension Project (Rural) of ICDDR,B. In total, deaths of 327 married adults aged 15-59 years, during January 1983-December 1987, were reviewed. The families of the deceased were followed up for five years after death. Factors, such as survival status of children, educational status of children aged 6-12 years, and out-migration status among adolescents aged 12-20 years in those families, was observed and recorded. A control group of 3,350 families experiencing no adult deaths was also followed up for five years. The health and socioeconomic impacts on children in both the groups five years after death of the adult were compared. The findings of the study showed that negative impact was more pronounced among the children of poor families, and the female children were most severely affected. Death of a father or a mother was associated with a higher rate of out-migration (especially marriage) of adolescent daughters. An adult death was associated with a significantly higher mortality risk of children during the five years following death of the adult. These child-mortality risks were significantly higher when an adult female died, and when the index child was a female and/or aged less than five years at the time of death of an adult. The children, aged 6-12 years, in families where a parent had died were significantly more likely to be uneducated and out-of-school after the death of a father or a mother compared to the children in families where neither of the parents had died. This finding remained valid even after controlling for the educational status of the parents who died and of those who did not die. Since the study used a limited number of independent variables and since there is a need to understand the specific reasons why such significant differences occurred, it is recommended to conduct a more in-depth qualitative study to know more clearly the nature and mechanisms of the socioeconomic and health impacts of death of an adult on the family and the society. PMID- 11855352 TI - Isolation of Salmonella enterica serotypes from children with diarrhoea in Calcutta, India. AB - The prevalence of Salmonella enterica serotypes, antimicrobial susceptibility, and phage typing of serovers were studied. Clinical presentations of the infected cases were also examined. The study was carried out during August 1993-September 1996 in and around Calcutta, India. In total, 1,025 faecal samples from hospitalized diarrhoeal children were screened for enteropathogens. Four S. enterica serotypes were identified in 157 (15.3%) cases as a single pathogen. S. enterica serotype Typhimurium was detected in 110 (70%) cases. S. Seftenberg, S. Infantis, and S. Virchow were detected in 28 (17.8%), 14 (8.9%), and 5 (3.2%) cases respectively. S. Typhimurium was isolated from 11 (3.2%) non-diarrhoeal control children. All of these children had acute watery diarrhoea, and 5% of them had severe dehydration, 40% had some dehydration, and 55% had no dehydration. Vomiting, fever, and diffused pain in abdomen were the associated presentations of these children. Most (95%) of them recovered with oral rehydration therapy only and without any antibiotics. In-vitro susceptibility testing showed that 120 of the 121 S. Typhimurium strains isolated from cases and controls were resistant to the commonly-used drugs. Thirteen of the 121 strains were phage-typeable and belonged to the phage type 193. However, no clinical or epidemiological significance could be established with these typeable strains. The findings of the study indicate that diarrhoeagenic Salmonella is one of the major pathogens causing diarrhoeal diseases in eastern India. PMID- 11855353 TI - Trends in caesarean section rates at a maternity hospital in Mumbai, India. AB - The steady rise in caesarean section rates is an emerging area of concern in mother-child healthcare and a matter of international attention, since the trend is no longer confined to western industrialized countries. Crude and caesarean section-related perinatal mortality and case-fatality rates may well serve as public-health indicators. Monitoring time-trends in caesarean section rates has been considered a useful approach in the recognition of this rapidly-changing health policy and in estimating the magnitude of this problem. The study examined the observed time-trends in caesarean section rates in relation to perinatal mortality rates and maternal case-fatality rates in a hospital setting in Mumbai, India, using 1957-1998 data on retrospective cohort. Both overall rates and those specific to type of delivery were assessed. During 1957-1998, the caesarean section rates in the Nowrosjee Wadia Maternity Hospital (NWMH) increased from 1.9% to 16%, with the most significant rise over the past decade. The perinatal mortality rate showed a significant reduction from 69 per 1,000 in 1957 to 36 per 1,000 in 1992 and remained steady in the 1990s despite the higher caesarean section rates. The caesarean section rate in the NWMH rose by almost 10-fold during 1957-1998. No improvement in perinatal outcome was observed beyond a caesarean section rate of 10%, but the perinatal mortality rate in caesarean births increased significantly due to a more liberal use of caesarean sections in preterm deliveries and those that yielded low-birth-weight babies. PMID- 11855355 TI - Determinants of health and nutritional status of rural Nigerian women. AB - This study was undertaken to determine the effects of socioeconomic and cultural factors on the health and nutritional status of 300 women of childbearing age in two rural farming communities in Enugu State, Nigeria. The women were engaged in farming, trading, and teaching. A cross-sectional survey was conducted using both qualitative and quantitative data-collection methods. The study involved focus group discussions (FGDs), interviews using a questionnaire, measurement of food/nutrient intake, assessment of activity patterns, anthropometry, and observations of clinical signs of malnutrition. The better-educated women had higher incomes than those with little or no education. Poor education was mainly attributed to lack of monetary support by parents (34%), marriage while in school (27%), and sex discrimination (21%). The teachers had significantly (p < 0.05) better health status, health and nutrition knowledge, food habits, nutrient intake, and self-concept, and adhered less to detrimental cultural practices. However, none of the women met their iron, riboflavin and niacin requirements. More cases of chronic energy deficiency were observed among the farmers (16%) and traders (13%) than among the teachers (5%). Generally, the women worked long hours with reported working hours (6-7 hours) being lower than the observed working hours (11 hours) for the traders and teachers. Income had a significant (p < 0.05) positive correlation with all nutritional variables, except vitamin C, age-at-marriage (r = 0.719), and nutrition knowledge (r = 0.601). Age-at-marriage had a positive correlation with body mass index (BMI) and all nutritional variables but was significant (p < 0.05) for protein (r = 0.362), calcium (r = 0.358), iron (r = 0.362), riboflavin (r = 0.364), and vitamin C (r = 0.476). Workload was negatively correlated with protein intake (r = 0.346; p < 0.05). Meal frequencies for more than 70% of the farmers and petty traders and 42% of the teachers were dependent on the availability of food in the household. Food taboos had no effect on their nutrient intake, since only 5-11% of women adhered to taboos. Although most women gave their children and husbands preference in food distribution, not much difference was found in the amount of food consumed by these women. The ratio of wife's portion to husband's was 1:1.4 for the farmers, 1:1.3 for the traders, and 1:1.2 for the teachers. FGDs revealed that sex discrimination in education prevailed where resources were limited. The results of the study suggest that the basic determinants of health and nutritional status of women are socioeconomic and cultural, education having a mediating or modifying influence on cultural practices. PMID- 11855354 TI - Sociodemographic, environmental and clinical risk factors for developing persistent diarrhoea among infants in a rural community of Egypt. AB - The study documents the incidence of persistent diarrhoea and its sociodemographic, household, environmental and clinical risk factors. One hundred and fifty-two newborns were followed for the first 12 months of life in Bilbeis, Egypt. The household of each participant was surveyed at baseline and was visited twice a week. The study infants experienced persistent diarrhoea at a rate of 0.51 episode/case per year. Development of persistent diarrhoea was associated with water storage in mud-containers (Odds ratio [OR] = 4.36, confidence interval [CI] 1.4-14.8), pump-water supply (OR = 3.5, CI 1.2-10.2), and absence of a latrine in the household (OR = 2.74, CI 1.01-7.38). Detection of faecal IgE (OR = 3.32, CI 1.0-10.9) and high stool frequency (OR = 2.95, CI 1.1-7.8) appeared as important clinical predictors for the onset of persistent diarrhoea. The incidence of persistent diarrhoea among young infants in Bilbeis, Egypt, was high. Sociodemographic, environmental and clinical parameters were important risk factors for the development of persistent diarrhoea. PMID- 11855356 TI - Shigellocidal properties of three Nigerian medicinal plants: Ocimum gratissimum, Terminalia avicennoides, and Momordica balsamina. AB - The prevalence of multidrug-resistant shigellae is an important concern in the treatment of shigellosis in many developing countries, and other therapies, including herbal agents, may provide an important alternative to antimicrobial agents. In this study, three Nigerian medicinal plants: Ocimum gratissimum, Terminalia avicennoides, and Momordica balsamina were investigated for their activities against multidrug-resistant Shigella species isolated from patients with bacilliary dysentery in Lagos. Decoctions of O. gratissimum and concoctions of O. gratissimum and T. avicennoides at crude concentration of 3,000 micrograms/mL markedly inhibited the growth of all isolates tested. Zones of inhibition indicating susceptibilities of the organisms varied from 18.3 to 21.5 mm for Shigella dysenteriae, 15.3 to 16.3 mm for S. flexneri, 18.8 to 19.3 mm for S. sonnei, and 16.5 mm for S. boydii. Except S. flexneri, minimum inhibitory concentration and minimum bactericidal concentration revealed a higher shigellocidal property of O. gratissimum/T. avicennoides concoction than other extracts in S. dysenteriae (300-515.6 vs 337.5-1,312.5 micrograms/mL), S. sonnei (309.4-543.8 vs 403.1-1,312.5 micrograms/mL), and S. boydii (243.8-337.5 vs 253 1,312.5 micrograms/mL). O. gratissimum showed a greater shigellocidal effect against the S. flexneri isolates, while extracts of M. balsamina possessed low shigellocidal potential. The results suggest that aqueous extracts of O. gratissimum and T. avicennoides as decoctions and concoctions could be useful in the treatment of shigellosis and should be clinically evaluated specially in Nigerian region. PMID- 11855357 TI - No isolation of Escherichia coli O157:H7 strains from faecal specimens of Turkish children with acute gastroenteritis. PMID- 11855358 TI - Effect of zinc supplementation on clinical course of acute diarrhoea. AB - This report summarizes the current knowledge on the effects of zinc supplementation on the management of acute diarrhoea. All published and unpublished studies on this topic, conducted in hospitals and in the community, were reviewed. Based on the results of this review, it is concluded that there is now enough evidence demonstrating the efficacy of zinc supplementation on the clinical course of diarrhoea, with regard to the severity and duration of the episode. However, the meeting also concluded that effectiveness studies to assess the feasibility, sustainability, and cost-effectiveness of different strategies for delivering zinc supplementation should be undertaken. PMID- 11855359 TI - American College of Toxicology: policy statement on the use of animals in toxicology. PMID- 11855360 TI - Danish validation of WHO Quality of Life Questionnaire. PMID- 11855361 TI - You're the flight surgeon: depth perception disorders. PMID- 11855362 TI - CMS Medicaid managed care proposal under close scrutiny. PMID- 11855363 TI - Economic boom over, state health budgets besieged. PMID- 11855364 TI - 16 X-chromosome STR loci frequency data from a German population. AB - Allele frequencies for 16 X-linked STRs, suitable for forensic purposes, were obtained from a sample of unrelated German individuals (male and female). The presented data show also repeat sequence structures and statistic parameters describing there information content. PMID- 11855365 TI - Profiler data for a Columbian population (Department of North Santander). PMID- 11855366 TI - The pathology of N-glycosylation--stay the middle, avoid the risks. PMID- 11855367 TI - My, what big eyes you have... PMID- 11855368 TI - Notice of triplicate publication. PMID- 11855369 TI - [Pain attacks causing muscle cramps]. PMID- 11855370 TI - [The "dental amalgam syndrome" - an environmental somatization Syndrome? A comparison between chronic carbon monoxide intoxication and illness related to dental amalgam]. AB - In 1940, during World War II, restrictions in import of petroleum products to Sweden necessitated the use of producer gas in motor traffic. In the following years, the incidence of acute carbon monoxide intoxications raised steeply. However, many patients with minor but longstanding exposition to producer gas exhibited a neurastenic syndrome (fatigue, headaches and vertigo) thought to be specific. In Stockholm, an epidemic of this syndrome can afterwards be traced to the personal conviction of an internist who also had an important influence on various authorities, leading to a forceful campaign to the public about the dangers of using producer gas. After some years, the frequency and even the existence of a chronic carbon monoxide intoxication was called in question and at the end of the war that diagnosis lost its actuality. In Sweden, oral galvanism attributed to dental amalgam was discussed in mass media in the 1970s, not least by evidence given by some well-known personalities. In the 1980s, the frequency of illness attributed to dental amalgam increased to an important epidemic. The question of the dangers of mercury released from amalgam fillings is still an important issue of debate among dentists and physicians, although the majority remains sceptical. Also medical authorities have found little evidence of the importance of dental amalgam toxicity. A patients organisation, Tandvardsskadeforbundet, seems to have played a significant part in the acceptance of the syndrome among laymen. Thus, various psychosocial factors seem to have played a role in both syndromes which could thus be conceived as environmental somatization syndromes. PMID- 11855371 TI - [Campaign in order to attract nursing students]. AB - During the late 1940s there was a shortage of nurses in Denmark. The Danish Council of Nurses (DSR) initiated a campaign in order to attract nursing students. This and other campaigns show the dilemmas within the nursing profession in this period. At the same time discussions on the future of the nursing profession took place and ministerial committees worked on the establishment of a countrywide uniform training for nurses. The campaigns highlighted on one side the traditions in the nursing profession, i.e. the feminine ability for care, which is based upon the feminine distinctive character. On the other side the campaigns pointed towards the future nursing profession, which is based upon professionalism and knowledge based upon schooling. PMID- 11855372 TI - [Ophthalmic tuberculosis, especially in Denmark]. AB - The contagiousity of tuberculosis was demonstrated by Carl J. Salomonsen in 1877, injecting human caseous material in rabbit eyes, causing granulomatous iritis. It was five years before Robert Koch found the bacterium: Mycobacterium tuberculosis (MT). Marius Tscherning as professor emeritus from the University of Copenhagen, injected bovine MT and sanocrysin (Molgaard's goldderivate) into rabbit eyes, causing corneal phlyctenules, but no ophthalmic reaction developed with only MT or sanocrysin alone (1928-29), not published). Emil Frandsen in 1959 examined 113 BCG vaccinated persons, who developed benign phlyctenules in 11%, 6-8 weeks after vaccination, and some cases of chronic iridocyclitis and chorioiditis. In 1158 non-vaccinated, some with serious tuberculosis, Frandsen found serious phlyctenules, chronic iridocyclitis, chorioiditis (especially disseminated) and periphlebitis, but not acute iridocyclitis of tuberculous origin. Phlyctenules and other TB-eye-diseases are described from Greenland 1926-76. Ophthalmic tuberculosis is still an important issue, also in Denmark today. PMID- 11855373 TI - [Hannover and Ibsen: two Danish anatomists in the middle of the 19th century. Were they enemies?]. AB - Adolph Hannover (1814-94), Danish anatomist and pathologist, introduced the microscopy to the medical research in Denmark. He published several papers on anatomy and pathology, among others Om Mikroskopets Bygning og dets Brug (1847) which was translated to several European languages, among others to English On the structure of the microscope and its use. As a jew he never succeeded in obtaining a post at the University of Copenhagen, Ib Pedersen Ibsen (1801-62) was educated at the Academia Chirurgorium Regia, the college of surgeons in Copenhagen, and he never became a scholarly anatomist, writing one paper only, published after his death. He was a competent anatomist and an appreciated teacher. The two anatomists competed for a post as lecturer in anatomy at the University of Copenhagen. Ibsen obtained the job after a public competition. The Museum of Medical History in Copenhagen is in possession of Ibsen's own copy of Hannover's dissertation on the cartilage, the muscles and the nerves of the external ear. In this copy Ibsen by hand has written some critical and unkind remarks against Hannover. Was this a sign of hostility? We conclude that they hardly were friends, but that the language between colleagues at the university could be rather harsh at that time. PMID- 11855374 TI - [The opening of the first real sick-hospital in Copenhagen: Kommunehospitalet]. AB - On the 19th, September 1863 the first real sick-hospital, Kommunehospitalet, opened in Copenhagen. This paper contains aspects of the temporary results and reflections from a interdisciplinary investigation between a pharmacist, a doctor and a ethnologist of the conditions at the opening the hospital. The investigation is based on the case records of the 89 patients who were recorded as admitted to the hospital on the opening day. The case records contain information not only of the diagnosis and treatments but offer the possibility to elucidate aspects of every day life both inside and outside the hospital, especially among the working-class and the poor who constitute the majority of the patients. Among the elucidated aspects is, that the patients are placed in the common ward as much according to their social status as to the diagnosis. Most of the admitted patients suffered from veneral diseases even though this category of patients had caused serious hesitation regarding the damage they could make to the reputation of and wish to create a new image for the hospital. PMID- 11855375 TI - [The blindness in the literature-Jose Saramago: blindness and Albert Bang: the blind witness]. AB - Two novels with different aspects of blindness seen through the doctors eyes. The Portuguese Nobel-prize winner Jose Saramago's story of a city struck by an epidemic of "white blindness", where the truth is what we cannot bear to see. The Danish author and unskilled labourer Albert Bang's (synonym with Karl E. Rasmussen) crime novel describes a blind or pretend to be blind butcher, who is a witness to a murder. Both novels are lyric, thought-provoking and insightful. PMID- 11855376 TI - Hereditary localized pruritus. AB - A dominant, possibly x-linked hereditary form of localized pruritus with onset in the third decade is described. PMID- 11855377 TI - Inhibitory effects of antifungal proteins on human immunodeficiency virus type 1 reverse transcriptase, protease and integrase. AB - A variety of antifungal proteins were isolated from seeds of leguminous plants including French bean, cowpea, field bean, mung bean, peanut and red kidney bean. They were assayed for ability to inhibit human immunodeficiency virus type I (HIV 1) reverse transcriptase, protease and integrase, enzymes essential to the life cycle of HIV-1 . It was found that the cowpea beta-antifungal protein had a high potency in inhibiting HIV-1 protease and HIV-1 integrase. Cowpea alpha-antifungal protein was potent in inhibiting HIV-1 reverse transcriptase and HIV-1 integrase. Peanut antifungal protein was characterized by a high inhibitory activity against HIV-1 integrase and an intermediate potency in inhibiting HIV- I reverse transcriptase and HIV- I protease. French bean thaumatin-like protein expressed low HIV- I protease inhibitory activity and red kidney bean lectin inhibited HIV- I integrase by only a very small extent. Antifungal proteins from the field bean and mung bean had an intermediate potency in inhibitory HIV-1 protease and integrase. However, mung bean antifungal protein was not capable of inhibiting HIV-1 reverse transcriptase. The results indicate that nearly all leguminous antifungal proteins examined were able to inhibit HIV-1 reverse transcriptase, protease and integrase to some extent. PMID- 11855378 TI - The role of osteoclast differentiation in aseptic loosening. AB - The major cause of orthopaedic implant loosening is thought to be accelerated osteoclastic bone resorption due to the action of cytokines produced in response to phagocytosis of implant-derived wear particles. This accelerated osteoclastic bone resorption could be due to increases in any of the following processes: recruitment of osteoclast precursors to the local microenvironment, differentiation of precursors into mature multinucleated osteoclasts. activation of mature osteoclasts, and/or survival of osteoclasts. Our studies have focused on differentiation and survival to complement work by others who have focused on recruitment of precursors and activation. Taken together, our studies and those of other investigators provide strong evidence that increased recruitment of osteoclast precursors and their subsequent differentiation play major roles in wear particle-induced osteolysis. In contrast, increased osteoclast activation and survival appear to play minor roles. These studies suggest that development of therapeutic interventions that reduce either recruitment or differentiation of osteoclast precursors would improve the performance of orthopaedic implants. PMID- 11855379 TI - EMG activity of trunk muscles and torque output during isometric axial rotation exertion: a comparison between back pain patients and matched controls. AB - Abnormal patterns of trunk muscle activity could affect the biomechanics of spinal movements and result in back pain. The present study aimed to examine electromyographic (EMG) activity of abdominal and back muscles as well as triaxial torque output during isometric axial rotation at different exertion levels in back pain patients and matched controls. Twelve back pain patients and 12 matched controls performed isometric right and left axial rotation at 100%, 70%, 50% and 30% maximum voluntary contractions in a standing position. Surface EMG activity of rectus abdominis, external oblique, internal oblique, latissimus dorsi, iliocostalis lumborum and multifidus were recorded bilaterally. The primary torque in the transverse plane and the coupling torques in sagittal and coronal planes were measured. Results showed that there was a trend (P = 0.08) of higher flexion coupling torque during left axial rotation exertion in back pain patients. Higher activity for external oblique and lower activity for multifidus was shown during left axial rotation exertion in back pain group when compared to the control group. In right axial rotation, back pain patients exhibited lesser activity of rectus abdominis at higher levels of exertion when compared with matched controls. These findings demonstrated that decreased activation of one muscle may be compensated by overactivity in other muscles. The reduced levels of activity of the multifidus muscle during axial rotation exertion in back pain patients may indicate that spinal stability could be compromised. Future studies should consider these alternations in recruitment patterns in terms of spinal stability and internal loading. The findings also indicate the importance of training for coordination besides the strengthening of trunk muscles during rehabilitation process. PMID- 11855380 TI - Telomerase and telomeres: from basic biology to cancer treatment. AB - The limited capacity to divide is one of the major differences between normal somatic cells and cancerous cells. This 'finite life span' of somatic cells is closely linked to loss of telomeric DNA at telomeres, the 'chromosome caps' consisting of repeated (7TAGGG) sequences., In more than 85% of advanced cancers, this telomeric attrition is compensated by telomerase, 'the immortality enzyme', implying that telomerase inhibition may restore mortality in tumor cells. This review discusses the progress in research on the structure and function of telomeres and the telomerase holoenzyme. In addition, new developments in telomere/telomerase targeting compounds such as antisense oligonucleotides and G quadruplex stabilizing substances, but also new telomerase expression-related strategies such as telomerase promoter-driven suicide gene therapy and telomerase immunotherapy will be presented. It will be discussed how these data can be implemented in telomerase-directed therapies. PMID- 11855381 TI - Evidence for D-aspartyl-beta-amyloid secretase activity in human brain. AB - Alzheimer disease (AD) is characterized neuropathologically by the presence of senile plaques that are composed of the amyloid-beta protein (Abeta). Abeta is an insoluble extracellular deposit consisting of 39-43 amino acids that is cleaved from a larger precursor amyloid-beta-precursor protein (beta-APP). It has been shown that Abeta proteins extracted from amyloid cores of neuritic plaques contain isomerized and/or racemized Asp residues. Therefore, we hypothesized that a specific secretase (s) may exist in the human brain that can cleave a beta-APP peptide bond containing D-Asp at position 1 of the Abeta protein. In the present study, we report data to support the existence of a putative membrane-bound D beta-secretase that can cleave between L-Met-D-Asp at the 1 position of the Abeta with a pH optimum in the neutral pH range. The specific enzyme activity of soluble extracts from AD samples was 22% higher compared to age-matched controls. PMID- 11855382 TI - Blocked gap junctional coupling increases glutamate-induced neurotoxicity in neuron-astrocyte co-cultures. AB - Gap junctional communication is likely one means by which neurons can endure glutamate cytotoxicity associated with CNS insults (i.e. ischemia). To examine this neuroprotective role of gap junctions, we employed gap junctional blockers to neuronal and astrocytic co-cultures during exposure to a high concentration of extracellular glutamate. Co-cultures were treated with the blocking agents carbenoxolone (CBX; 25 microM), 18alpha-glycyrrhetinic acid (AGA; 10 microM), vehicle or the inactive blocking analogue glycyrrhizic acid (GZA; 25 microM). Twenty-four hours following the insult, cell mortality was analyzed and quantified by the release of lactate dehydrogenase (LDH) into the media, the cells' inability to exclude propidium iodide, and terminal dUTP nick end labeling (TUNEL). Measurement of LDH release revealed that the glutamate insult was detrimental to the co-cultures when gap junctions were blocked with CBX and AGA. Based on propidium iodide and TUNEL labeling, the glutamate insult caused significant cell death compared to sham vehicle and mortality was amplified in the presence of CBX and AGA. Since blockers were not themselves toxic and did not affect astrocytic uptake of glutamate, it is likely that blocked gap junctions lead to the increased glutamate cytotoxicity. These findings support the hypothesis that gap junctions play a neuroprotective role against glutamate cytotoxicity. PMID- 11855383 TI - Altered tubulin and neurofilament expression and impaired axonal growth in diabetic nerve regeneration. AB - Cytoskeletal protein expression in sensory neurons and sciatic nerve axonal growth were examined in type 1 diabetic BB/Wor rats after sciatic nerve crush injury. Diabetic male rats were subjected to sciatic nerve crush at 6 wk of diabetes. L4 and L5 dorsal root ganglia (DRG) mRNA expression of low and medium molecular weight neurofilaments (NF-L, NF-M), betaII- and betaIII-tubulin as well as protein expression of NF-L, NF-M, and beta-tubulin were examined at various time points following crush injury and compared with age- and sex-matched non diabetic BB/Wor rats. Steady state mRNA expression of NF-L, NF-M, betaII- and betaIII-tubulin were decreased in diabetic DRG. NF-L and NF-M proteins were also decreased in DRG of uncrushed diabetic animals. After crush injury, betaII- and betaIII-tubulin mRNA were upregulated in control animals at day 2 and day 6, respectively, and beta-tubulin protein showed similarly increased expression after crush injury, while such upregulations did not occur in diabetic animals. Conversely, mRNA and protein expressions of NF-L, NF-M were downregulated to a lesser extent in diabetic animals compared to control rats. These changes were associated with impaired axonal elongation and caliber growth of regenerating fibers in diabetic rats. We propose that upregulation of tubulin has a negative feedback on NF expression in response to nerve injury, as seen in control rats. The absence of this upregulation in diabetic animals may impair its regulatory effect on NF expression and contribute to perturbed nerve regeneration seen in diabetic nerve. PMID- 11855384 TI - From the Centers of Disease Control and Prevention. Recommended childhood immunization schedule--United States, 2002. PMID- 11855385 TI - From the Centers of Disease Control and Prevention. Progress toward interrupting indigenous measles transmission--region of the Americas, January-November 2001. PMID- 11855386 TI - From the Centers of Disease Control and Prevention. Evaluation of postexposure antibiotic prophylaxis to prevent anthrax. PMID- 11855387 TI - Of course patients should be told. PMID- 11855388 TI - The surgeon is only one factor. PMID- 11855389 TI - Rapid assessment of chest pain. Chest pain clinics may be one step forward, two steps back. PMID- 11855390 TI - Wrong biochemistry results. Information on incidents with consequences for health should be collected centrally. PMID- 11855391 TI - Wrong biochemistry results. Interdepartmental cooperation may help avoid errors in medical laboratories. PMID- 11855392 TI - Monitoring safety of over the counter drugs. Over the counter medicines must be treated like all other medicines. PMID- 11855393 TI - Daily regimen and compliance with treatment. Concordance respects beliefs and wishes of patients. PMID- 11855394 TI - Daily regimen and compliance with treatment. Literature before 1980 should not have been ignored. PMID- 11855395 TI - Screening for diabetes in general practice. Population screening for diabetes is cost effective. PMID- 11855396 TI - Screening for diabetes in general practice. Workload studies as well as clinical trials should be considered when drawing up guidelines. PMID- 11855397 TI - National guidance and allocation of resources. Economics has both strengths and weaknesses in health resource allocation. PMID- 11855398 TI - Is my care a good example? PMID- 11855399 TI - Enhancing the state of play in children's nursing. AB - Play is recognised as an essential component of a child's life. Playing with children in hospital can aid nurses in assessing, communicating and providing nursing care. Play workshops can facilitate nursing students' understanding of the importance and benefits of play. Play can benefit children, health carers and parents in the hospital setting. Play workshops encourage children's nurses to view play as an integral part of their role. PMID- 11855400 TI - Information giving can be painless. AB - Pain is an inevitable consequence following day surgery, but with effective regular pain relief minimal discomfort can be achieved. An information leaflet produced by a pain nurse specialist in collaboration with a multidisciplinary team ensures that comprehensible and non-contradictory information is given to parents. With the increased trend towards paediatric day surgery parents are significant contributors to their child's care in the postoperative period. Written information on pain relief after discharge can help parents make decisions about their child's pain management. PMID- 11855401 TI - Great expectations? Parental participation in care. AB - A structured questionnaire was used to compare the views of 80 parents and nurses examining expectations of parental participation in care. Nurses and parents shared many expectations. However, there were instances when parents expected a different and sometimes greater level of participation. A lack of negotiation between parents and nurses with regard to expected roles resulted in parents often feeling unable to express their desired level of participation. Both parties indicated that if parents received greater guidance and support, it would be possible for their expectations to be met more adequately. PMID- 11855402 TI - Ways of working: CCNs and chronic illness. AB - The study explored the role/skills used by CCNs caring for children with chronic illness. A participant inquiry paradigm was used and data were generated using semi-structured interviews supported by extensive field notes. All the CCNs described the special relationship with the children and their families which was characterised by deep understanding of current and future needs. The CCNs emphasised 'ways of working' which involved high degrees of trust, flexibility, support, reflexivity and empowerment. CCNs help families regain, maintain and develop control over their lives. PMID- 11855403 TI - Quality counts: auditing day-surgery services. AB - Clinical audit is a valuable tool in the drive towards quality. Day-case surgery is designed to keep children within their families. Parents of children having day-case surgery must be adequately informed and supported. A telephone call following day-case surgery appears to give parents effective support. Children experience few problems immediately following day-case surgery. PMID- 11855404 TI - Consent in children: legal and ethical issues. AB - Children's wishes are often not respected. The law does not support children's involvement in consent. Nurses can be influential in supporting patients during the process of consent. Decisions may be made based upon ethical principles, which tend not to respect individual rights. PMID- 11855405 TI - How Amie feels within. PMID- 11855406 TI - STEPS: the National Association for Children with Lower Limb Abnormalities. PMID- 11855407 TI - Proportional counter as neutron detector. AB - A technique to separate out the dose, and lineal energy spectra of neutrons and charged particles is described. It is based on using two proportional counters, one with a wall, and the other with similar characteristics but wall made from a non-hydrogen containing material. Results of a calibration in a neutron field are also shown. PMID- 11855408 TI - Overview of ground-based neutron measurements applicable to GCR and SPE transport through shielding materials in space. PMID- 11855409 TI - Neutron measurements using bubble detectors--terrestrial and space. AB - Like all other radiation monitors currently used in the space program, the bubble detector (which has flown on several missions) was adapted from a technology that was developed for terrestrial radiation. Bubble detectors are the most recent technology for applications in personal neutron dosimetry. They are now regarded as a mature technology and are used in many countries as a neutron dosimeter of record with approval from the respective regulatory authorities. Extensive type testing and QA of bubble detectors has been done by numerous groups, many of these to show that bubble detectors meet national radiation protection requirements prior to their acceptance as a dosimeter of record. In fact, it has been stated "Only bubble detectors achieve a sufficiently low detection threshold (to meet ICRP-60 recommendations)..." (Portal and Dietze, 1992). PMID- 11855410 TI - Development of the fiber neutron monitor for the energy range 15-100 MeV on the International Space Station (ISS). AB - In order to investigate the space environment, a new neutron monitor has been prepared. The sensor consists of scintillation fibers (FIB) and will be on board the exposed facility of the Japanese Experimental module (Kibo) of the International Space Station (ISS). The sensor is one of the instruments which measures the particle and plasma environment around the ISS. The sensor will be used by the cosmonauts who work on the ISS, to help them avoid a strong particle radiation which may arrive soon. We believe that such an alert is necessary for the cosmonauts to prevent unnecessary radiation from the sun. Furthermore, the sensor will be very useful in understanding the particle acceleration mechanism at the solar surface. The design of the Engineering Model (EM) was finished and its properties were investigated. This report discusses these results. PMID- 11855411 TI - An application of artificial neural intelligence for personal dose assessment using a multi-area OSL dosimetry system. AB - Significant advances have been made in recent years to improve measurement technology and performance of phosphor materials in the fields of optically stimulated luminescence (OSL) dosimetry. Pulsed and continuous wave OSL studies recently carried out on alpha-Al2O3:C have shown that the material seems to be the most promising for routine application of OSL for dosimetric purposes. The main objective of the study is to propose a new personal dosimetry system using alpha-Al2O3:C by taking advantage of its optical properties and energy dependencies. In the process of the study, a new dose assessment algorithm was developed using artificial neural networks in hopes of achieving a higher degree of accuracy and precision in personal OSL dosimetry system. The original hypothesis of this work is that the spectral information of X- and gamma-ray fields may be obtained by the analysis of the response of a multi-element system. In this study, a feedforward neural network using the error back-propagation method with Bayesian optimization was applied for the response unfolding procedure. The validation of the proposed algorithm was investigated by unfolding the 10 measured responses of alpha-Al2O3:C for arbitrarily mixed photon fields which range from 20 to 662 keV. PMID- 11855412 TI - Development of personal neutron dosemeters at the PTB and first measurements in the space station MIR. AB - A passive neutron dosemeter with thermoluminescence and etched-track detectors was used in the space station MIR in 1995 and 1997 and during some shuttle flights to MIR. High neutron doses of about 200 microSv d-1 were measured with track detectors, while the contribution of protons to the track density was estimated to be small. An active personal dosemeter based on silicon diodes providing a direct readout, improved sensitivity and spectrometric properties is proposed for additional monitoring. Firstly, measurements with a prototype were performed in the stray radiation fields of the CERN-EU Reference Radiation Facility. When mounted on a phantom at different positions, both the passive and the active dosemeter provide the directional distribution of neutrons via an unfolding procedure. This can be helpful for a better understanding of the complex radiation field in space and for comparisons with calculations. PMID- 11855413 TI - Russian measurements of neutron energy spectra on the Mir orbital station. AB - Results of the experiments on neutron energy spectra measurements within broad energy range from 5 x 10(-7) to 2 x 10(2) MeV aboard the Mir orbital station and equivalent neutron dose estimation are presented. Four measurement techniques were used during the experiments. The shape of spectra and their absolute values are in good agreement. According to those experiments, an equivalent neutron dose depends upon effective shielding thickness and spacecraft mass. The neutron dose mentioned is comparable with that of ionizing radiation. Neutron flux levels measured aboard the Mir station have shown that a neutron spectrometer involving broad energy range will be used within the radiation monitoring systems in manned space flights. PMID- 11855414 TI - Real-time measurement of low-energy-range neutron spectra on board the space shuttle STS-89 (S/MM-8). AB - We have developed a real-time, Bonner Ball-type (neutron energy range is from thermal to 15 MeV) neutron spectral measurement system (Bonner Ball Neutron Detector (BBND)) for use on board the International Space Station (ISS). From measurements taken inside STS-89 (S/MM-8), we successfully distinguished neutrons from protons and other particles in a mixed radiation field; a task hitherto considered difficult. Although the experimental period was short, only 3.5 days (January 24-27, 1998), we were able to obtain energy spectral data and the Earth's neutron dose-equivalent map for the ISS orbital conditions (altitude 400 km, orbit inclination angle 51.6 degrees). A method for calculating the neutron energy spectrum and compensating for the particle interaction with the sensors is also described in detail. PMID- 11855415 TI - Simulation of Space Shuttle neutron measurements with FLUKA. AB - FLUKA is an integrated particle transport code that has enhanced multigroup low energy neutron transport capability similar to the well-known MORSE transport code. Gammas are produced in groups but many important individual lines are specifically included, and subsequently transported by the main FLUKA routines which use a modified version of EGS4 for electromagnetic (EM) transport. Recoil protons are also transported by the primary FLUKA transport simulation. The neutron cross-section libraries employed within FLUKA were supplied by Giancarlo Panini (ENEA, Italy) based upon the most recent data from JEF-1, JEF-2.2, ENDF/B VI, JENDL-3, etc. More than 60 different materials are included in the FLUKA databases with temperature ranges including down to cryogenic temperatures. This code has been used extensively to model the neutron environments near high-energy physics experiment shielding. A simulation of the Space Shuttle based upon a spherical aluminum equivalent shielding distribution has been performed with reasonable results. There are good prospects for extending this calculation to a more realistic 3-D geometrical representation of the Shuttle including an accurate representation of its composition, which is an essential ingredient for the improvement of the predictions. A proposed project to develop a combined analysis and simulation package based upon FLUKA and the analysis infrastructure provided by the ROOT software is under active consideration. The code to be developed for this project will be of direct application to the problem of simulating the neutron environment in space, including the albedo effects. PMID- 11855416 TI - Neutron dosimetric measurements in shuttle and MIR. AB - Detector packages consisting of thermoluminescence detectors (TLD), nuclear emulsions and plastic track detectors were exposed at identical positions inside MIR space station and on shuttle flights inside Spacelab and Spacehab during different phases of the solar cycle. The objectives of the investigations are to provide data on charge and energy spectra of heavy ions, and the contribution of events with low-energy deposit (protons, electrons, gamma, etc.) to the dose, as well as the contribution of secondaries, such as nuclear disintegration stars and neutrons. For neutron dosimetry 6LiF (TLD600) and 7LiF (TLD700) chips were used both of which have almost the same response to gamma rays but different response to neutrons. Neutrons in space are produced mainly in evaporation and knock-on processes with energies mainly of 1-10 MeV and up to several 100 MeV, respectively. The energy spectrum undergoes continuous changes toward greater depth in the attenuating material until an equilibrium is reached. In equilibrium, the spectrum is a wide continuum extending down to thermal energies to which the 6LiF is sensitive. Based on the difference of absorbed doses in the 6LiF and 7LiF chips, thermal neutron fluxes from 1 to 2.3 cm-2 s-1 are calculated using the assumption that the maximum induced dose in TLD600 for 1 neutron cm-2 is 1.6 x 10(-10) Gy (Horowitz and Freeman, Nucl. Instr. and Meth. 157 (1978) 393). It is assumed that the flux of high-energy neutrons is at least of that quantity. Tissue doses were calculated taking as a mean ambient absorbed dose per neutron 6 x10(-12) Gy cm2 (for a10 MeV neutron). The neutron equivalent doses for the above-mentioned fluxes are 52 micro Gy d-1 and 120 micro Gy d-1. In recent experiments, a personal neutron dosimeter was integrated into the dosimeter packages. First results of this dosimeter which is based on nuclear track detectors with converter foils are reported. For future measurements, a scintillator counter with anticoincidence logic is under development. PMID- 11855417 TI - HZETRN: neutron and proton production in quasi-elastic scattering of GCR heavy ions. AB - The development of transport models for radiation shielding design and evaluation has provided a series of deterministic computer codes that describe galactic cosmic radiation (GCR), solar particle events, and experimental beams at particle accelerators. These codes continue to be modified to accommodate new theory and improvements to the particle interaction database (Cucinotta et al., 1994, NASA Technical Paper 3472, US Government Printing Office, Washington DC). The solution employed by the heavy-ion transport code HZETRN was derived with the assumption that nuclear fragments are emitted with the same velocity as the incident ion through velocity conserving nuclear interactions. This paper presents a version of the HZETRN transport code that provides a more realistic distribution of the energy of protons and neutrons emitted from GCR interactions in shields. This study shows that the expected GCR dose equivalent is lower than previously calculated for water shields that are less than 110 g cm-2 thick. Calculations of neutron energy spectra in low Earth orbit indicate substantial contributions from relativistic neutrons. PMID- 11855418 TI - Estimation of neutron and other radiation exposure components in low earth orbit. AB - The interaction of high-energy space radiation with spacecraft materials generates a host of secondary particles, some, such as neutrons, are more biologically damaging and penetrating than the original primary particles. Before committing astronauts to long term exposure in such high radiation environments, a quantitative understanding of the exposure and estimates of the associated risks are required. Energetic neutrons are traditionally difficult to measure due to their neutral charge. Measurement methods have been limited by mass and weight requirements in space to nuclear emulsion, activation foils, a limited number of Bonner spheres, and TEPCs. Such measurements have had limited success in quantifying the neutron component relative to the charged components. We will show that a combination of computational models and experimental measurements can be used as a quantitative tool to evaluate the radiation environment within the Shuttle, including neutrons. Comparisons with space measurements are made with special emphasis on neutron sensitive and insensitive devices. PMID- 11855419 TI - Validation of the galactic cosmic ray and geomagnetic transmission models. AB - A very high-momentum resolution particle spectrometer called the Alpha Magnetic Spectrometer (AMS) was flown in the payload bay of the Space Shuttle in a 51.65 degrees x 380-km orbit during the last solar minimum. This spectrometer has provided the first high statistics data set for galactic cosmic radiation protons, and helium, as well as limited spectral data on carbon and oxygen nuclei in the International Space Station orbit. First measurements of the albedo protons at this inclination were also made. Because of the high-momentum resolution and high statistics, the data can be separated as a function of magnetic latitude. A related investigation, the balloon borne experiment with a superconducting solenoid spectrometer (BESS), has been flown from Lynn Lake, Canada and has also provided excellent high-resolution data on protons and helium. These two data sets have been used here to study the validity of two galactic cosmic ray models and the geomagnetic transmission function developed from the 1990 geomagnetic reference field model. The predictions of both the CREME96 and NASA/JSC models are in good agreement with the AMS data. The shape of the AMS measured albedo proton spectrum, up to 2 GeV, is in excellent agreement with the previous balloon and satellite observations. A new LIS spectrum was developed that is consistent with both previous and new BESS 3He observations. Because the astronaut radiation exposures onboard ISS will be highest around the time of the solar minimum, these AMS measurements and these models provide important benchmarks for future radiation studies. AMS-02 slated for launch in September 2003, will provide even better momentum resolution and higher statistics data. PMID- 11855420 TI - Radiation dose from reentrant electrons. AB - In estimating the crew exposures during an extra vehicular activity (EVA), the contribution of reentrant electrons has always been neglected. Although the flux of these electrons is small compared to the flux of trapped electrons, their energy spectrum extends to several GeV compared to about 7 MeV for trapped electrons. This is also true of splash electrons. Using the measured reentrant electron energy spectra, it is shown that the dose contribution of these electrons to the blood forming organs (BFO) is more that 10 times greater than that from the trapped electrons. The calculations also show that the dose-depth response is a very slowly changing function of depth, and thus adding reasonable amounts of additional shielding would not significantly lower the dose to BFO. PMID- 11855421 TI - Measurements of LET-distribution, dose equivalent and quality factor with the RRMD-III on the Space Shuttle Missions STS-84, -89 and -91. AB - Dosimetric measurements on the Space Shuttle Missions STS-84, -89 and -91 have been made by the real-time radiation monitoring device III (RRMD-III). Simultaneously, another dosimetry measurement was made by the Dosimetry Telescope (DOSTEL) on STS-84 and by the tissue-equivalent proportional counter (TEPC) on STS-91. First, the RRMD-III instrument is described in detail and its results summarized. Then, the results of DOSTEL and TEPC are compared with those of the RRMD-III. Also, the absorbed doses obtained by TLD (Mg2SiO4) and by RRMD-III on board STS-84 and -91 are compared. PMID- 11855422 TI - Regulating the market for human eggs. AB - This essay provides a rationale for a regulated market for human oocytes. Although the commodification of human oocytes raises important moral concerns, these concerns do not justify laws banning commerce in human eggs. Given the burgeoning ART industry and the growing oocyte market, the most prudent course of action is to develop regulations for the human oocyte market that are designed to protect and promote important social values, such as health, safety, liberty, and respect for human life. Other responses, such as banning the sale of eggs altogether or allowing donors to be compensated only for their services, would either create a black market or would lead to corruption and abuse. Society still needs to debate specific rules and policies that should govern the human egg market, but further discussion of that important task is best left to legislative bodies and other commentators. PMID- 11855423 TI - New federalwide assurance for protection of human subjects. PMID- 11855424 TI - Instruction in responsible conduct of research (RCR). PMID- 11855425 TI - Research misconduct and human subjects. PMID- 11855426 TI - IRB duties and genetic testing. PMID- 11855427 TI - Widespread IRB-related compliance problems at research institution. PMID- 11855428 TI - Protecting forcibly institutionalized mental patients from unwanted drug therapy: a fourth amendment analysis. PMID- 11855429 TI - Pederasty, blood shedding and blood smearing: men in search of mommy's feared powers. PMID- 11855430 TI - Childhood and fantasies of medieval mystics. PMID- 11855432 TI - If it is a girl, cast it out: infanticide/exposure in Ancient Greece. PMID- 11855431 TI - Begriming the unsullied: a theme driving historical action. PMID- 11855433 TI - The evolution of childrearing. PMID- 11855434 TI - Surrendering the self: the reduction of identity to identification with the aggressor. The cases of Hermann Goring and Rudolf Hess. PMID- 11855435 TI - The phantom Jew: from spoiled identity to genocide. PMID- 11855436 TI - Descartes's Regulae, mathematics, and modern psychology: "the Noblest example of all" in Light of Turing's (1936) On computable Numbers. AB - There are surprisingly strong connections between the philosophy of mind and the philosophy of mathematics. One particular important example can be seen in the Regulae (1628) of Descartes. In "the noblest example of all," he used his new abstract understanding of numbers to demonstrate how the brain can be considered as a symbol machine and how the intellect's algebraic reasoning can be mirrored as operations on this machine. Even though his attempt failed, it is illuminating to explore it because Descartes launched 2 traditions--mechanistic philosophy of mind and abstract mathematics--that would diverge until A. Turing (1936) approached symbolic reasoning in a similar "symbol machine-existence proof" way. Descrates's and Turing's thought experiments, which mark the beginning of modern psychology and cognitive science, respectively, indicate how important the development of mathematics has been for the constitution of the science of mind. PMID- 11855437 TI - The power of a musical instrument: Franklin, the Mozarts, Mesmer, and the glass armonica. AB - In 1761 Benjamin Franklin invented the armonica (often referred to as the glass harmonica), an instrument designed to simplify the playing of the musical glasses. The instrument immediately became popular and inspired compositions by Wolfgang Mozart, who had the opportunity to hear and play one at the house of Franz Anton Mesmer. Armonica music was used by Mesmer in his seances, because he felt it could promote healing by propagating a mystical fluid that he called animal magnetism through the body. After Mesmer's theories were debunked by a highly respected panel of scientists, the armonica fell out of vogue. Because Franklin was on the panel that examined the discredited mesmerism, he indirectly contributed to his own invention's demise. PMID- 11855438 TI - The cult of experiment: the Psychological Round Table, 1936-1941. AB - The Psychological Round Table (PRT) was an important yet anomalous society founded by young American experimental psychologists. Although it was secretive, it encouraged contact among young experimental psychologists. Although it was motivated by perceived inequities in professional psychology, it was governed by an autocratic minority. And although it was self-consciously informal and irreverent, it clandestinely used formal selection procedures. These aspects of the PRT are documented and explained (counter to L. Benjamin, 1977) as manifestations of the PRT's devotion to psychological experimentation, specially understood. Secrecy, specifically, served not to protect PRT participants from senior colleagues but to create an environment free of political and professional influences and thus taken to be well suited to the vetting of experiments. In this regard, the PRT is distinguished from more familiar scientific societies. PMID- 11855439 TI - My experience with a second brain aneurysm. PMID- 11855440 TI - Washington state CNSs are the first NACNS affiliate group of the millennium. National Association of Clinical Nurse Specialists. PMID- 11855441 TI - When a staff member is causing conflict: scapegoat, a management problem or an opportunity for change in culture? PMID- 11855443 TI - Research--what's important about CNSs doing it? PMID- 11855442 TI - Mental health consultation in a nursing home. AB - As the world's population ages, increasing numbers of people can anticipate spending their latter years in long-term care settings. Many of these nursing home residents will also present psychiatric illnesses as primary or secondary diagnoses. The resulting behavioral problems may present challenges to nursing staff that they are ill-prepared to meet. This article illustrates the application of the Blake and Mouton consultation model to a Veterans Administration (VA) nursing home situation by a team of psychiatric mental health nurse specialists. The consultation is described and interpreted in terms of the Blake and Mouton model. The focal conflicts addressed in the consultation included issues of morale/cohesion, power/authority, and norms/standards. Interventions used were acceptant, prescriptive, confrontation, and theories/principles. The model provided a useful structure for conceptualizing and organizing assessment and intervention in the consultation situation. PMID- 11855444 TI - Comparing CNS and NP role activities: a replication. AB - Since the early 1980s, there has been discussion about combining the clinical nurse specialist (CNS) and nurse practitioner (NP) roles. The implications of such a merger have generated much concern and controversy. This study replicates Williams and Valdivieso's 1992 descriptive study comparing the CNS and NP roles to gain more insight into current implementation of role activities. A demographic questionnaire, the Advanced Nursing Practice Survey, and the Work Activities Checklist were used to poll 610 CNSs and NPs practicing in Minnesota. The findings of the study revealed significant differences in CNS and NP role activities. Analysis of the replication and original study findings suggested no evolution toward a blended CNS/NP role. These data imply that while the roles have similarities they continue to have areas of unique practice. Results of the study support the continued existence of the two advanced practice roles. PMID- 11855445 TI - Responses and concerns of healthcare providers to medication errors. AB - This descriptive, correlational study examined the responses and concerns of healthcare professionals about making medication errors and estimated patient harm from such errors. A systematic random sample of nurses, pharmacists, and physicians (N = 402) completed a self-report survey about a medication error they judged to be serious. Respondents were guilty, nervous, and worried about the error. They feared for the safety of the patient, disciplinary action, and punishment. A few subjects indicated that they never reported the errors. The most frequent symptoms associated with errors were neurologically based. The injury suffered by patients was not severe overall according to the harm scales. Weak correlations were found for the harm scales and responses and concerns. The authors suggest a supportive environment for the provider following an error and continuous quality improvement efforts to eliminate system-based errors. PMID- 11855446 TI - Reunion. PMID- 11855447 TI - Adverse drug event reporting and advanced-nursing practice. AB - Adverse drug events are undesirable effects suspected of being caused by drugs. They account for 1.7-11% of all hospital admissions. Occurrence in hospitalized patients is 1-44%, yet less than 10% are reported to national agencies. Reporting can lead to regulatory action, including label changes and drug withdrawal. These actions result in increased public safety and decreased healthcare costs. Nurses account for less than 10% of reporters. Advanced practice nurses are in a key position to report; those with prescriptive authority have a public health responsibility to do so. Master's-level nursing programs in the US were surveyed. Results showed that about 75% of responding schools promoted awareness of reporting, but only about half taught how to report and what to report. Adverse drug event reporting is a significant professional issue that requires not only formal education but also ongoing education and support for advanced practice nurses and staff. PMID- 11855448 TI - CPT coding and Medicare reimbursement issues. PMID- 11855449 TI - A model for the future. Integration of the bachelor of nursing honours degree with the graduate nurse program. AB - We describe an innovative integrated study pathway at the School of Nursing, Flinders University in which students can complete a Bachelor of Nursing Honours degree and Graduate Nurse Program in one year. PMID- 11855450 TI - What's best? Clinical indicators of quality in residential aged care facilities. AB - This paper reports on the views of 36 registered nurses (RNs) in Phase One of a three stage study, Quality of Care for Residents in Aged Care Facilities. Case studies were conducted in nine residential aged care facilities and data were collected from informants using semi-structured interviews, participant observation and document analysis and review. Of the 36 RNs, almost three quarters (n=26) provided care at the bedside and ten held managerial positions. Each volunteered to participate. When asked to nominate the major clinical indicators of high quality residential aged care, all 26 RNs who worked at the bedside stated that the absence of decubitis ulcers was the pre-eminent measurable factor. While five managerial RNs also mentioned low rates of pressure ulcers, only two ranked it as the most important clinical indicator of high quality care. Hydration management was the clinical indicator nominated most frequently by managerial nurses. The one indicator of equal importance to both groups, but for different reasons, was that of poly pharmacy. The differences in priorities between each group were statistically significant. PMID- 11855451 TI - Development of clinical assessment criteria for postgraduate nursing students. AB - This paper explores the development and implementation of specific criteria to determine the level of clinical performance of postgraduate nursing students during the first year of a Master of Nursing course. The authors describe two commonly used clinical skill assessment tools and identify limitations of these tools for postgraduate nursing students. As a result of these limitations, Clinical Assessment Criteria (CAC) utilising the framework of Benner (1984) was developed. Inherent within the CAC is four levels of clinical nursing performance, which enable the nurse teacher and student to monitor the progression from novice to proficient levels of practice within a specialty area. Following a successful pilot study, the CAC was incorporated into clinical assessments in nine specialty postgraduate courses. Furthermore, the framework developed for the CAC can also be integrated into a variety of professional development domains. PMID- 11855452 TI - Specialist nurse: an investigation of common and distinct aspects of practice. AB - Over the past decade the discipline of nursing has been reviewing its practice, especially in relation to specialty areas. There has been an appreciation by nursing leaders that specialisation brings with it concerns related to a disuniting effect on the discipline and a fragmentation of nursing's traditional generalist practice. Accompanying these concerns is a debate over what is a specialty and how to define a specialist. This qualitative study drew upon a constructivist methodology, to explore how nurses, working in specialty areas, define and give meaning to their practice. Three groups of nurses (n=20) from the specialty of critical care were interviewed using a focus group technique. The data were analysed to build constructions of specialty practice. A distinct and qualitative difference was recognised in the practice behaviours of nurses working in the specialty area. The qualitatively different practice behaviours have been identified as 'nursing-in-a-specialty' and 'specialist nurse'. Two constructions emerged to differentiate the skill behaviours, these were 'practice' and 'knowledge'. The specialist nurse practices were based on two distinct types of practice, that of 'discretion' and 'incorporation'. 'Knowledge' was constructed as a synthesis of propositional and practice knowledge. By Carrie Fairweather and Glenn Gardner. PMID- 11855454 TI - Acute pain and its management. PMID- 11855453 TI - On donkeys, ingratiates, thinkers and...clinical development units. AB - Clinical Development Units celebrate nurses as thinkers and nursing as a highly skilled professional activity. They are typified by the exploration and development of therapeutic, consumer-focused and research-based practice and participatory governance. Establishing and maintaining them, therefore, can be problematic in health care facilities typified by centralism, autocracy and nurses' disempowerment. In this type of facility nurses are treated like donkeys and/or ingratiates. What CDUs are, the standards nurses in them strategically and systematically pursue and their achievements to date will be discussed. In addition,some sociopolitical impediments to their development will be identified together with recommendations for their successful establishment and maintenance. PMID- 11855455 TI - Palliative care. PMID- 11855457 TI - The caring ethic. PMID- 11855458 TI - Collaboration with psychiatrists in Connecticut. PMID- 11855459 TI - Statutory and regulatory issues for clinical nurse specialist (CNS) practice: ensuring the public's access to CNS services. AB - The practice environment of healthcare in the United States is fluid, dynamic, and sometimes chaotic. Statutory, regulatory, and credentialing requirements are critical elements of this changeable environment affecting clinical nurse specialists (CNSs) and other advanced practice nurses (APNs). CNSs are estimated to comprise approximately 54,000 APNs, based on the 2000 National Sample Survey of Registered Nurses. Although statutes and regulations related to APNs are reviewed regularly, there has not been a critical analysis of statutes and regulations specifically governing CNS practice in the United States. This lack is largely due to the absence of a national association devoted to CNSs before 1995. However, recently there has been a resurgence of interest and need for CNS services. Therefore, it is particularly important that any statutory and/or regulatory barriers to CNS practice be removed. In 1998, the National Association of Clinical Nurse Specialists Legislative and Regulatory Committee embarked on a critical analysis of state statutes and regulations governing CNS practice. All 50 state boards of nursing and the District of Columbia board of nursing were requested to send copies of their statutes and regulations governing CNS practice. Responses were received from 48 states. This article summarizes the state of regulation of CNS practice and identifies significant barriers to CNS practice that should be systematically addressed. The patchwork quilt of CNS regulation that varies from state to state results in over-restrictive and underinclusive provisions that preclude ease of reciprocity and deprive the public full access to CNS services. Recommendations for addressing the barriers are made. PMID- 11855460 TI - National Association of Clinical Nurse Specialists model statutory and regulatory language governing clinical nurse specialist practice. PMID- 11855461 TI - First new management for poisonous snakebites in more than 50 years is now available. Biotechnology product provides safe and effective therapy for poisonous snake bites. PMID- 11855462 TI - National healthcare coalition releases student focus group results. First step in advertising campaign to fight nursing shortage. PMID- 11855463 TI - Congresswoman Lois Capps speaks out on the nursing shortage. PMID- 11855464 TI - Mentors and mentoring. PMID- 11855465 TI - Where have all the experts gone? PMID- 11855466 TI - Community analysis for health planning with vulnerable populations. AB - This article describes how advanced practice nurses used a theory-based community health analysis process to examine the healthcare services most needed and used by an adult public housing community and to examine the satisfaction of the community with those services. The goal of the project was to determine if the identified needs would be amenable to an alternative healthcare delivery model, such as an academic nursing clinic. Data were collected using the Health Needs and Health Status Survey, Center for Epidemiological Studies-Depression scale, interactive participant interviews, key informant interviews and observations. Community residents (N = 242) were typically unmarried, middle-aged, and low income and had multiple chronic physical illnesses, serious and persistent mental illnesses, and/or disabilities. Despite the high identified service need, use of preventative or routine healthcare services was low, with residents using emergency services inappropriately or delaying care until requiring more expensive intensive healthcare services. PMID- 11855467 TI - Complexity, critical thinking and intellectual virtues. PMID- 11855468 TI - Drawing together hope: 'listening' to militarised children. AB - Art can be used as both a therapeutic tool and a communication tool. Art has been used with refugees/militarised children but has tended to focus on their pathology. Art in this study was used to explore children's hopes for their future. Children talk more when they are/have drawn what they are trying to describe. Art can be used to understand children's world-view and explore their beliefs and opinions. PMID- 11855469 TI - Nurturing a child's spirituality. AB - Nurturing a child's spirituality should be an integral part of holistic care. The concept of spirituality is linked to the child's cognitive, social, psycho-sexual and moral development. Knowledge of childhood spirituality can help to support children coping with traumatic life-events. The expression of beliefs and feelings that encompass spirituality can be facilitated through literature and music and through other strategies. Educators need to empower professionals to have the awareness, emotional resources and skills to ensure that they can be spiritually supportive PMID- 11855470 TI - Simple febrile convulsions: evidence for best practice. AB - Febrile convulsions are common in early childhood and may be the first sign of an illness. Witnessing the convulsion and experiencing the subsequent admission is extremely stressful for parents. Children's nurses need to identify the evidence for best practice in the management of febrile convulsions. Parent education is an essential component of the management of care. PMID- 11855471 TI - Anthropology and children's nursing. AB - Children's nurses have become more involved in the provision of culturally sensitive care. There has been a search for a more meaning centred (or hermeneutic) method in exploring children's worlds. The possible relevance of anthropology as an approach to both the culture of children's nursing and the cultures in which children's nurses find themselves is explored. PMID- 11855472 TI - Reflections of a novice researcher. AB - Nurses can benefit from active participation in research activities. Involvement in research is not just an academic pursuit but is essential for effective, quality care. Nurses should actively seek the support of colleagues and research support structures to enable involvement in research. Clinical nurses are capable of making an important research contribution. PMID- 11855473 TI - Enteral feeding: a change in practice. AB - There is an unacceptably high incidence of malnutrition in PICU. The delivery of nutritional support requires a structured approach. Early enteral feeding has been shown to reduce the stress response. Implementing a change in practice involves identification of resources, objectives and motivation for change. A randomised retrospective audit of the change in practice did demonstrate the value of earlier commencement of enteral feeding. Informal interviews with practitioners suggested improved practice and knowledge. PMID- 11855474 TI - Munchausen syndrome by proxy abuse. AB - The difficulties in diagnosing Munchausen Syndrome by Proxy Abuse (MSbPA) are examined. The perpetrators of MSbPA are usually the victim's primary care giver. The warning signs of MSbPA include unexplained, prolonged and extraordinary illness with the signs and symptoms only being evident when the suspected perpetrator is present. Confrontation, exclusion and covert video surveillance are all possible routes to gaining further evidence. The problems associated with covert video surveillance are explored. PMID- 11855475 TI - Giving our little princess her new voice. Part two. PMID- 11855476 TI - REACT: giving depth to short lives. PMID- 11855477 TI - Human subjects protection and advanced practice nurses. PMID- 11855478 TI - Gapology--the study of gaps in the nursing knowledge base. PMID- 11855479 TI - Perceptions about advance directives by nurses in a community hospital. AB - Despite the legal mandate for hospitals to comply with the Patient Self Determination Act and recommendations by the American Nurses' Association for nurses to advocate for the participation of patients in end-of-life decisions, nurses' compliance has been less than enthusiastic. This study used an exploratory descriptive design and a 10-item self-reported questionnaire, which included both multiple-choice and open-ended questions. This study examined nurses' knowledge and comfort with the implementation of the Patient Self Determination Act. An analysis of this research shows that two major themes emerged: a need for more education involving advance directives and a desire to have other healthcare workers involved in informing patients about advance directives. PMID- 11855480 TI - Content validation of the Medication Error Worksheet. AB - Clinical nurse specialists use a variety of preexisting instruments to measure and describe health-related concepts. It is important for clinical nurse specialists to know how to evaluate the content validity of potentially useful instruments. This study assessed the content validity of the Institute for Safe Medication Practice's Medication Error Worksheet. The worksheet is used as a questioning framework to guide data collection processes when beginning analysis of a medication error. Although the worksheet has been valuable to the Institute for Safe Medication Practice staff, its content validity has not been determined. Content validity methods included expert validation and a review of the related literature. Results support the validity of the Medication Error Worksheet and suggest that this worksheet is a comprehensive tool that may be helpful when exploring the circumstances of medication errors and when analyzing medication use systems. Results were shared with the Institute for Safe Medication Practice staff to improve the accuracy of the worksheet. PMID- 11855481 TI - Outcome evaluation: an essential component for CNS practice. 1999 [classical article]. PMID- 11855483 TI - Ethical dilemmas: how can the clinical nurse specialist provide ethical expertise? PMID- 11855482 TI - Pharmacologic management of patients undergoing conscious sedation. PMID- 11855484 TI - Understanding pity in O'Faolain's Up the Bare Staircase. PMID- 11855485 TI - Statutory and regulatory recognition for clinical nurse specialists in Oregon. AB - Clinical nurse specialists (CNSs) in Oregon initiated the process of achieving statutory and regulatory recognition several years ago. Throughout this process, specific phases of activity and events helped CNSs to identify what was required to achieve this goal. The resulting lessons learned are shared in this report. Statutory recognition of CNSs in Oregon occurred in 1999, and the administrative rules for CNS practice were published in 2001. These administrative rules delineate the CNS scope of practice and other aspects of CNS practice consistent with national standards. PMID- 11855486 TI - Learning "how to" conduct clinical research from others' experiences. PMID- 11855487 TI - The challenges of conducting clinical research: one research team's experiences. AB - Conducting clinical research is an exciting and a rewarding endeavor. Each project is unique and, because of this uniqueness, research studies require considerable planning. Regardless of all the upfront planning, challenges will occur throughout the life of the study. Clinical research projects are rife with obstacles that range from the lack of administrative and physician support to subject attrition. Some of the challenges, such as subject dropout, are not unexpected, whereas other unanticipated issues can blindside a research team. This case study examines several such challenges experienced in one longitudinal study and presents the solutions engineered by the research team to keep the project on track. PMID- 11855488 TI - Noncompliance or human nature? PMID- 11855490 TI - APNs-influencing practice through research. PMID- 11855489 TI - A CNS-managed diabetes foot-care clinic: a descriptive survey of characteristics and foot-care behaviors of the patient population. AB - Lower extremity lesions are the primary cause of hospitalization for people with diabetes, resulting in enormous personal and financial costs. This study used a survey designed to describe the characteristics and foot-care behaviors of people with diabetes who attended a clinical nurse specialist managed foot-care clinic. Forty-eight patients who received care at the participating foot-care clinic completed a 21-item multiple-choice questionnaire designed to determine the presence of foot pathology and foot-care behaviors. Most of the patients were between 65 and 74 years of age, had concurrent illnesses, and had four or more primary care visits per year. Although 69% had existing foot pathology, only 44% reported inspecting their feet daily and only 54% reported that their primary care provider examined their feet on each visit. Twenty-five percent reported going barefoot sometimes and eight percent would either treat a foot lesion themselves or wait for it to get better. PMID- 11855491 TI - Activities of the APN to enhance unpartnered elders self-efficacy after myocardial infarction. AB - Unpartnered elders recovering from a myocardial infarction find themselves with limited support to contend with their physical and emotional recovery. The advanced practice nurse (APN) can play an instrumental role in improving the health outcomes of these elders. Detailed description of the activities of the APN that affect patient outcomes in current randomized clinical trials have been inadequate in published reports. Therefore, the purpose of this article is to describe the activities of an APN in a nurse-coached intervention study for unpartnered elders post-myocardial infarction. Thematic analysis from the subject logs and narratives from the APN identified the four themes of patient education, validation/feedback, encouragement/support, and problem solving as the unique activities of the APN. These activities enhanced the recovery of these vulnerable unpartnered elders after a myocardial infarction. PMID- 11855492 TI - Exercise program effects on women with fibromyalgia syndrome. AB - The purpose of this study (evaluation) was to examine the effects of an exercise program on 13 women with physician-diagnosed fibromyalgia syndrome (FMS). Participants engaged in exercise for 60 minutes each session. Group 1 (N=7) was in a 3-day-per-week program for 12 months, and group 2 (N= 6) was in a 3-day-per week program for six months. Group 3 (N= 3) consisted of three participants from Group 1 who participated for six additional months past the 12-month period (total--18 months). Group 3 attended five sessions per week during the six additional months. All participants engaged in aerobic and resistance training. Information was collected on physical fitness, psychosocial, and FMS symptom variables. A majority of the participants appeared to experience a positive outcome on numerous measures of physical fitness, psychosocial factors, and FMS symptoms. Interview data support results. The 13 participants gained various benefits from the exercise program and functioned the same or better outside of the program. Implications for advising FMS patients relative to exercise are given for clinical nurse specialists. PMID- 11855493 TI - The science and art of nursing or so what? PMID- 11855494 TI - Nurses as scientists. PMID- 11855495 TI - A practical approach to the process of measurement in nursing. PMID- 11855496 TI - Sexuality. PMID- 11855497 TI - Pharmacologic management of agitation in the older adult with dementia. PMID- 11855499 TI - User beware of information on the Internet. PMID- 11855500 TI - Solving Medicare reimbursement problems in Connecticut. PMID- 11855501 TI - A career ladder for the clinical nurse specialist: 'to be or not to be". PMID- 11855502 TI - Senior clinical nurse specialist pilot position in Hong Kong. AB - This article reports findings of a consultancy project commissioned by Hospital Authority, Hong Kong, to explore the scope of service, role competence, and contribution to patient service of senior-position clinical nurse specialists. Three clinical nurse specialists were selected for this project. The consultant guided and monitored effects of initiatives proposed by each clinical nurse specialist. The evaluation strategies were dependent on the nature of the projects proposed by the senior clinical nurse specialist. Data were collected through nonparticipant observation and interviews to address the overall consultancy project aims. This study shows that the role components of clinical nurse specialists were common across different countries. The value of the clinical nurse specialist to initiate strategies to promote cost-effective and quality care were confirmed, but the career development of these specialists remains a concern in a healthcare environment with constraining resources. PMID- 11855503 TI - Power: its use and potential for misuse by nurse consultants. 1991. PMID- 11855504 TI - What will it take to reduce errors in healthcare settings? PMID- 11855505 TI - Nurse practitioiners (NPs) and clinical nurse specialists (CNSs) regarding the blending of these two roles. PMID- 11855506 TI - Access to complementary health care: why the NHS is the key. PMID- 11855507 TI - Shamanism as a healing paradigm for complementary therapy. AB - Any healing process--whether recovery from infection, physical trauma, or psychological distress--must entail the stimulation and direction of the body's own restorative functions. In former times these functions were called the vis mediatrix naturae. Arguably best articulated within traditional Chinese medicine (e.g. Reid 1993), many complementary therapies have identified this principle. The immune system is implicated in the operation of these healing processes, and immune system functions are modulated by both internal and external variables. External variables include the nature of the infection or trauma. Internal variables include the meaning of the illness to the patient or the patient's imagery surrounding the illness. It follows that any modulation of internal variables that increases immune function will therefore be highly beneficial in the healing process. Sometimes such modulation happens spontaneously, when it may be referred to as the placebo effect, or a good bedside manner, or spontaneous remission. Sometimes such modulation may be brought about intentionally either by the patient or by a therapist or healer. One body of technique for such modulation is shamanism, which pays particular attention to bridging the internal world of the patient to the external world where the problem originates. Shamanic practice is specifically focused on this healing task, and has its own toolkit of techniques for the modification of consciousness, the manipulation of imagery and meaning, and the generation of a healing milieu and therapeutic images from its mythic content. Early concerns about the mental health of shamanic practitioners are now thoroughly resolved (e.g. Stephen & Suryani 2000). Indeed, the relevance of shamanism to positive mental health is currently being explored (e.g. Money 1994, Singh 1999). Its relevance to social work (Voss et al. 1999) and to the near death experience (Green 1998) are also subjects of academic inquiry. The shamanic corpus exemplifies a healing paradigm that may also be used to understand the essential elements of healing, which underpin some established complementary therapies and some other healing modalities such as spiritual healing,'psychic' healing, spontaneous remission, and the placebo effect. The comparatively recent psychoneuroimmunological perspective appears to be congruent with and also to validate ancient shamanic healing technique. Both may share essential principles with complementary therapies and illuminate their essential healing processes. PMID- 11855508 TI - Is there a place for auricular therapy in the realm of nursing? AB - Several theories, such as the 'homuncular reflex theory' 'delta reflex theory', and 'meridian theory' point to the fact that the ear is related to all parts of the human body and internal organs. Being one of the approaches in traditional Chinese medicine, auricular therapy is a therapeutic method by which specific points on the auricle are punctured or pressed. Auricular therapy can activate meridians and collaterals, regulate the Qi and blood, help to achieve the balance between Yin and Yang status of internal organs, and is therefore suitable for treating many disorders of the body. Successful examples of previous studies using this therapy including insomnia, weight reduction, hypertension, treatment of addiction, and pain reduction. However, inconsistency in the treatment protocol among studies, or the use of combined therapies, makes it impossible to draw a strong causal relationship between this therapy and the treatment effect. More appropriate clinical trials are therefore necessary to understand in depth the therapeutic effect of auricular therapy. Ideally, these trials can take place in the context of nursing practice so as to explore the application of this therapy in the realm of nursing, and to enable nurses to make a more effective contribution to primary health care. PMID- 11855509 TI - What's wrong with bias? PMID- 11855510 TI - Complementary therapies and nursing models. AB - It is suggested that personal construct theory not only has the potential as a model for nursing, but also for complementary therapies, thus allowing complementary therapies or procedures from the therapies and nursing to blend, promoting a seamless integration that enables nurses to offer a wider range of interventions in their nursing care. By coming together in a therapeutic relationship using personal construct theory, it is possible for nurses to enhance the process of healing which is an integral part of nursing care. PMID- 11855511 TI - The scope and limitations of treatment. An interview with Ann Lett, Principle, British School--Reflex Zone Therapy of the Feet. AB - Ann Lett was born in Johannesburg South Africa and trained as a nurse in 1953, and subsequently as a midwife in London. In 1977 she was treated with reflex zone therapy following a road traffic accident, and subsequently pursued a career in reflex zone therapy by undertaking training with many European practitioners. In 1979 she trained with Hanne Marquardt in Germany, and in 1981 was asked to develop the British School of Reflex Zone Therapy at the National Hospital for Nervous Diseases in London. Shortly after this, she and Ms. Marquardt established schools of reflex zone therapy in Israel and Barcelona. Her book Reflex Zone Therapy for Health Professionals, published in 2000 by Churchill Livingstone (ISBN 0 443 060150), is the culmination of 23 years of expertise in reflex zone therapy, and 47 years of nursing experience. She is presently Principle, British School Reflex Zone Therapy of the Feet, Wembley Park, UK. In this interview, Ann discusses the scope and limitations of reflexology and how she sees this profession developing in the future. PMID- 11855512 TI - Honoring the cycles, in society and our personal lives. PMID- 11855513 TI - The integration of complementary therapies into conventional medicine focuses primarily upon symptomatic care of the body with scant attention given to any interrelationship of the spirit and mind upon health and well-being. AB - We are pleased to continue our series of debates addressing topical issues currently under scrutiny in the field of complementary medicine. In selected issues, two people are invited to argue the case for or against a specific statement. In certain instances the position taken may not necessarily reflect the personal views of the authors. They have been asked to take a specific stance in order to stimulate debate and reflection about particular aspects of complementary and alternative medicine. PMID- 11855514 TI - Emerging innovations: caring in action. Re-creating Nightingale: the pain and the passion. PMID- 11855515 TI - Thought field therapy. Interview by Denise Rankin-Box. AB - Thought field therapy (TFT), one of the new 'power therapies' uses the body's meridian system to facilitate emotional and psychological change. Developed by clinical psychologist Dr Roger Callahan over the last 22 years,TFT is now used internationally. Recent developments indicate that TFT has the potential to be used more widely in the field of health care. The editor recently interviewed Jo Cooper to find out more about this therapy, PMID- 11855516 TI - Been there, seen it, got the "Essence" tee shirt...? PMID- 11855517 TI - Parents' perceptions of information on immunisations. AB - This study sought to understand parents' experiences of deciding to have their child immunised. A qualitative method was used, involving semi-structured interviews with a purposive sample of 13 parents. Parents identified feelings associated with risk, including those of worry, fear, and responsibility. Influences affecting their decision-making, and ways in which they received information on immunisation, were also identified. Parents made suggestions for changes relating to information giving, discussion time, and the acknowledgement by professionals of parents' worry. Following this study, discussion within the PHCT has resulted in a practice protocol and immunisation template. PMID- 11855518 TI - Professional relationships: CCN's and primary care nurses. AB - Children's community nursing is a relatively new service to provide nursing care to children in the community. Collaboration between new services and established community services is necessary to ensure children receive care from the most appropriate service. The views of health visitors, school nurses, district nurses and children's community nurses were sought to provide insight into collaborative processes. Focus groups were used to obtain participants' views of professional relationships. Each professional group viewed their relationship with the children's community service through the context of their own service. Expectations of children's community nurses by professionals were different for each professional group. PMID- 11855519 TI - Bereavement education for childcare professionals. AB - Bereavement is a stressor that affects not only the child but also the whole family. Children need time and support to come to terms with their grief. Current evidence suggests that professionals feel ill equipped to deal with these issues. Inter-professional, multi-agency collaboration and support can provide effective, integrated and supportive services. A 12-day modular course was developed within the Exeter Community Trust and accredited by the University of Plymouth. PMID- 11855520 TI - Developing child and adolescent mental health services. AB - Twenty four projects nationwide received government funding to improve child mental health services. These initiatives are expected to deliver innovative changes and enhanced interprofessional working. This preliminary evaluation in Norfolk suggests early intervention can offer acceptable, accessible, and appropriate support for children and families. PMID- 11855521 TI - Health promotion strategy for adolescents' sexual behaviour. AB - Adolescents distinctive health problems, such as, risks attached to unprotected sex, are cause for concern to all children's nurses. Peers play a crucial role in the psychological and social development of most adolescents. Peer education can be used as a health promotion strategy with adolescents. PMID- 11855522 TI - AFASIC: unlocking speech and language. PMID- 11855523 TI - Diverse practitioners join at the heart of healing. PMID- 11855524 TI - The Alexander technique: its application in midwifery and the results of preliminary research into Parkinson's. AB - This article outlines the origins and theory of the Alexander Technique and how it is learnt. Recent research is listed and a description given of main presenting symptoms and medical problems of pupils coming for lessons. The paper also describes how the Alexander Technique is used by women in pregnancy and childbirth and explains why it is beneficial. The final section describes Parkinson's research results, mainly from a preliminary study designed some years ago to test a methodology for a randomized controlled trial. PMID- 11855525 TI - Complementary strategies in antenatal care. AB - This paper describes the complementary therapy antenatal clinic at Queen Mary's Hospital, Sidcup, Kent, UK, run by a midwife who uses a range of complementary therapies to treat women with, mainly, physiological symptoms of pregnancy. PMID- 11855526 TI - Baby massage classes and the work of the International Association of Infant Massage. AB - This article describes the role of the International Association of Infant Massage (IAIM) in promoting the importance of touch for the developing relationship between the parents and infants. Infants are unique personalities and it is important that the parent learns how to respond appropriately to the infant's needs. Infant massage classes can demonstrate and promote interaction with infants using eye-to-eye contact, recognizing infant cues, talking and singing, and responding in a loving and sensitive manner. The photographs are of an infant massage class held recently in Leicestershire. PMID- 11855527 TI - The effects of Healing Touch on the coping ability, self esteem and general health of undergraduate nursing students. AB - The study used a mixed methodology to investigate the effects of HealingTouch (HT) on the coping ability, self esteem and general health of undergraduate nursing students to: check the veracity of claims made by HT practitioners that the therapy is effective in reducing stress; monitor the effects of HTon participants' scores using instruments which measure coping, self esteem and general health; and compare objective data with subjective accounts relating to coping, self esteem and general health. Quantitative findings showed no effects of HT on the coping ability, self esteem and general health of first year students, but there were some slight effects for the third year students who were in the experimental group. Qualitative findings showed that all participants in the experimental group found the experience positive. However, conclusions may be drawn tentatively that HT is effective while acknowledging that this project's design may have been ineffective in measuring the effects of HT, as the questionnaire may not have tested the actual effects of the therapy, and objective measures may be inadequate for exploring subjective experiences such as HT. PMID- 11855528 TI - Working with survivors of torture in Sarajevo with Reiki. AB - While working as a nurse/therapist in Sarajevo, I had the opportunity to work in an experimental situation at a center for torture survivors. This was to see if the use of Reiki would have a beneficial effect on this type of patient. It involved a rethink on traditional Reiki hand positions, music, and the general set-up of the room being used. It was a challenge, and one I was delighted to have had. The people I worked with were wonderful, and the changes in them over the period were so positive. The staff at the Center were delighted; I was delighted; but so much more importantly, the patients were delighted. The ground has now been broken and hopefully will be considered in a positive light for other traumatized patients. PMID- 11855529 TI - 'Reflexology techniques are not an effective tool for symptom recognition or the diagnosis of medical conditions'. AB - We are pleased to introduce a new series of debates addressing topical issues currently under scrutiny in the field of complementary medicine. In selected issues, two people are invited to argue the case for or against a specific statement. In certain instances the position taken may not necessarily reflect the personal views of the authors. They have been asked to take a specific stance in order to stimulate debate and reflection about particular aspects of complementary and alternative medicine (CAM). PMID- 11855530 TI - The caring dance. AB - As nursing and health care practice enter the new Millennium, practitioners are increasingly urged to pay attention to evidenced based practice to justify what they do. Yet the truth is, that within the caring dance, practitioners need to connect with more ancient sources of wisdom. Failure to do so leads to a life out of balance and a failure to dance well and fulfil the fundamental role of being a nurse. The paper draws exclusively on the work of Blackwolf and Gina Jones, as an example of such ancient wisdom to inform and inspire the caring dance. PMID- 11855531 TI - Clinical nurse specialist as facilitator of interdisciplinary collaborative program for adult sickle cell population. AB - This article describes the evolution, implementation, and outcomes of a collaborative interdisciplinary inpatient program designed to improve the quality of healthcare provided to adult patients with sickle cell disease. Efforts by the clinical nurse specialist were directly responsible for improved patient education, more efficacious treatment of painful crises, and increased specialty consultation. Staff and patient satisfaction with the program have been very encouraging, and a city-wide initiative that embraces a comprehensive approach to the global care of this special needs population has been designed and is currently being implemented. PMID- 11855532 TI - Living and partly living. PMID- 11855533 TI - The CNS: making cents out of costly outliers through nursing care. PMID- 11855534 TI - Development of a program to manage costly outliers. AB - The advanced practice nurse group devised a method to identify clinical variables of the elderly patients with multisystem failure requiring complex nursing care referred to as outlier. Outliers in this program were defined as patients whose hospital charges were greater than $50,000 with a length of stay greater than the primary diagnostic related group designated. Once criteria were identified, nursing strategies were developed to monitor the elderly patient, implement interventions, and evaluate patient outcomes. The goals of this program were to identify who the outliers might be prior to becoming outliers and to manage their nursing care early in their hospital course, attempting to match resource requirements with resource availability. PMID- 11855535 TI - Ethical decision making: integral to the role of the advanced practice nurse. PMID- 11855536 TI - The role of CNSs in promoting elderly patients' autonomy in long-term institutions: problems and implications for nursing practice and research. AB - Autonomy has become an important focus within healthcare. Elderly patients, however, are often denied the right to independent decision making on grounds of incompetence and incapacity. In long-term institutions there are several ways in which the clinical nurse specialist can promote the autonomy of elderly patients. This article provides an overview of the concept of autonomy and discusses some of the problems in its realization in long-term nursing institutions. It also outlines strategies for supporting autonomy in elderly patients and concludes with a discussion of the implications of autonomy to nursing practice and research. PMID- 11855537 TI - Ambulatory esophageal pH monitoring: technique, interpretations, and clinical indications. AB - Gastroesophageal reflux disease (GERD) is the most common esophageal disorder and perhaps among the most prevalent conditions seen in the primary care setting. The clinical manifestations of GERD, typical or atypical, such as noncardiac chest pain, respiratory or ear, nose, and throat symptoms, result from the reflux of gastric contents into the esophagus. Thus the clinical spectrum is wide and requires accurate diagnosis. Ambulatory 24-hr esophageal pH monitoring is not useful in all patients suspected to have GERD. This review describes the technique of ambulatory 24-hr esophageal pH monitoring, the interpretation of findings, and clinical applications of this test. PMID- 11855538 TI - Slow fluid infusion alters esophageal tone before onset of esophageal contraction in humans. AB - We investigated whether esophageal tone was altered by fluid infusion before the onset of esophageal contraction. In nine healthy volunteers, intraluminal esophageal pressure and esophageal pH were measured with a three-channel solid state transducer and microglass electrodes, respectively. Saline or 0.1 N hydrochloric acid was infused at 0.25 ml/sec 5 cm above the lower esophageal sphincter. The effect of atropine (80 microg/kg, intravenously) was also studied. Infusion of either saline or hydrochloric acid suddenly increased the midesophageal intraluminal pressure (8.2-8.4 mm Hg) and decreased the pressure more distally (8.0-8.5 mm Hg) with a latency of 23 sec. There were no significant differences between the effects of saline and hydrochloric acid. Atropine significantly (P < 0.05) lessened the midesophageal increase in pressure, but only slightly inhibited the distal decrease. Atropine did not alter the latency. Esophageal tone was altered by slow fluid infusion. This previously unknown physiological response may be a mechanism for storing liquid in the esophageal body until the start of contractions for clearance. PMID- 11855539 TI - Hiatal hernia and acid reflux frequency predict presence and length of Barrett's esophagus. AB - One third of the general population may experience reflux symptoms, yet only a small fraction of patients with gastroesophageal reflux disease (GERD) have Barrett's esophagus. The aim of the present study was to compare the characteristics of GERD patients with and without Barrett's esophagus and identify potential risk factors for the appearance of Barrett's esophagus in reflux disease. Outpatients from a gastroenterology clinic who underwent upper gastrointestinal endoscopy, esophageal manometry, and 24-hr pH monitoring were recruited into a case-control study. A total of 256 case subjects with endoscopically and histologically proven Barrett's esophagus were compared to a control group of 229 subjects with nonerosive reflux disease. As compared to nonerosive reflux disease, Barrett's esophagus was strongly associated with more reflux episodes. Barrett's esophagus occurred more frequently among subjects with hiatus hernia and among subjects who consumed large amounts of alcohol or cigarettes. Frequent reflux episodes, hiatus hernia, smoking, and alcohol consumption were also risk factors for an increased length of Barrett's mucosa. Total esophageal mucosal acid contact time at pH < 4 was a significant risk factor for the length but not the presence of Barrett's esophagus. Intake of aspirin or NSAIDs was similar in patients with and without Barrett's esophagus. In conclusion, in comparison with nonerosive reflux disease, Barrett's esophagus is characterized by risk factors usually indicative of severe types of GERD. Mechanisms in addition to acid reflux must contribute to the development of Barrett's esophagus. PMID- 11855540 TI - Control of nocturnal gastric acidity: a role for low dose bedtime ranitidine to supplement daily omeprazole. AB - In some patients, proton pump inhibitors do not abolish nocturnal gastric acidity and additional evening antisecretory medication may be required. In 16 subjects with chronic heartburn, 24-hr gastric and esophageal pH were measured at baseline and again after six days of 20 mg omeprazole alone at 08:00 hr followed by placebo, 75 mg ranitidine, or 20 mg omeprazole at 22:00 hr. Integrated acidity was calculated from the cumulative, time-weighted mean acid concentrations (derived from pH values for each second). Baseline integrated gastric acidity increased progressively over 24 hr, whereas integrated esophageal acidity increased only until 22:00 hr. Morning omeprazole nearly abolished 24-hr esophageal acidity and significantly decreased overall gastric acidity but did not abolish nocturnal gastric acidity. Adding evening ranitidine or omeprazole nearly eliminated the nocturnal increase in gastric acidity. Integrated acidity was more sensitive than time pH < 4 in assessing gastric and esophageal acidity as well as their inhibition by omeprazole and ranitidine. In conclusion, integrated acidity provides novel information regarding the synergy of omeprazole plus ranitidine. Adding low-dose ranitidine helps control nocturnal gastric acidity that can occur with conventional omeprazole administration. Although the heartburn patients in the present study had nocturnal gastric acidity without accompanying nocturnal esophageal acid reflux, other patients who do have nocturnal esophageal reflux might profit from addition of bedtime ranitidine or another gastric antisecretory agent. PMID- 11855541 TI - Stem cell factor expressed in human gastric mucosa in relation to mast cell increase in Helicobacter pylori-infected gastritis. AB - We previously reported mast cell increases in H. pylori gastritis. To determine the mechanism, we investigated the kinetics of mast cells and mast cell growth factor (stem cell factor, SCF) in H. pylori-positive and -negative gastric mucosa. Biopsy specimens from 12 H. pylori-negative and 28 positive subjects were examined. Sections were stained for mast cells, proliferating cell nuclear antigen (PCNA), and SCF. Densities of mast cells, PCNA-positive mast cells, and SCF-positive cells were significantly greater in H. pylori-positive than negative subjects. SCF was expressed in mast cells and fibroblasts. The density of SCF-positive fibroblasts increased in H. pylori-positive gastritis and decreased after cure of infection. SCF mRNA was detected in H. pylori-positive gastric mucosa. Fibroblasts isolated from the normal gastric mucosa expressed SCF mRNA after incubation with H. pylori water extract. SCF may be one of the factors for mast cell increase. Fibroblasts may participate in mast cell increase and inflammation in H. pylori infection. PMID- 11855543 TI - Seroprevalence of H. pylori infection and symptoms of upper gastrointestinal tract disease in two groups of health-care workers. AB - The purpose of this study was to determine the seroprevalence of Helicobacter pylori and risk factors for infection in gastrointestinal endoscopy personnel and those caring for developmentally disabled individuals. We administered a structured, one-page questionnaire. Helicobacter pylori status was determined serologically. In all, 107 health-care workers participated. Direct patient contact, exposure to gastrointestinal secretions, history of peptic ulcer, and acid-suppressing medicine use were more frequent among those who worked with the developmentally disabled. Seroprevalence of Helicobacter pylori was 24% in endoscopy personnel and 47% among those who cared for the developmentally disabled (P = 0.02). Bed-sharing with a sibling during childhood, working with the developmentally disabled, and direct patient contact were associated with Helicobacter pylori infection; exposure to patients' gastrointestinal secretions was not. We conclude that those caring for developmentally disabled individuals have a higher risk of occupational acquisition of Helicobacter pylori infection than endoscopy personnel. PMID- 11855542 TI - Effect of rebamipide in treatment of Helicobacter pylori-associated duodenal ulcer: attenuation of chemokine expression and nitrosative damage. AB - Production of cytokines along with increased activity of nitric oxide synthase has been implicated as one of the contributing mechanisms of Helicobacter pylori mediated gastroduodenal diseases. We aimed to evaluate the effect of rebamipide in treating Helicobacter pylori-associated duodenal ulcers in terms of cytokine production and nitrosative damage of the gastric mucosa. In patients with duodenal ulcers, rebamipide or placebo were given randomly after eradication. Mucosal cytokine production was measured by enzyme linked immunoassay, and nitrotyrosine immunoexpression was measured by immunohistochemistry. The inflammatory activity and degree of neutrophil infiltration were graded accordingly. The mucosal production of RANTES, interleukin-8, and TNF-alpha showed a significant decrease after eradication in patients with rebamipide after treatment. The nitrotyrosine immunoreactivity of gastric epithelium was significantly decreased in the rebamipide group. Rebamipide treatment after eradication resulted in a significant reduction in chemokine production along with nitrotyrosine immunoexpression in Helicobacter pylori-associated duodenal ulcers. PMID- 11855544 TI - [14C]Urea breath test is not sensitive for detection of acute Helicobacter pylori infection in rhesus monkeys (Macaca mulatta). AB - The urea breath test is sensitive and specific for detection of chronic infection with H. pylori. We sought to determine the sensitivity of the [14C]urea breath test for detection of acute H. pylori infection using experimentally infected rhesus monkeys. Eighteen monkeys were inoculated with H. pylori. Serial [14C]urea breath tests and cultures of gastric biopsies were performed before and up to 10 weeks after inoculation. Cultures from all 18 monkeys were positive for H. pylori at each time point. The sensitivity of the [14C]urea breath test increased systematically from 43% at two weeks after inoculation up to 93% at 10 weeks after inoculation. Quantitative cultures of H. pylori showed a tendency to decline over time following inoculation. We conclude that the [14C]urea breath test is not sensitive for detection of acute H. pylori infection in rhesus monkeys until 10 weeks after inoculation. While this may reflect a gradual increase in bacterial load that was not detected by limited sampling, our data are not consistent with this hypothesis. PMID- 11855545 TI - Unusual duodenal duplication cyst associated with partial gastric diverticulum in a middle-aged woman: are they congenital or acquired? PMID- 11855546 TI - Synchronous proximal polyps and cancer in patients with polyps detected at sigmoidoscopy: results of a single, rural-based sigmoidoscopy clinic. AB - The prevalence of polyps and cancer in the proximal colon among patients who have polyps detected on sigmoidoscopy was determined in a large rural referral hospital in north central Pennsylvania. Eleven thousand one hundred sixty patients underwent sigmoidoscopy between 1991 and 1997. Polyps were detected in 709 patients. Five hundred twenty-three patients who had a polyp at sigmoidoscopy and full colonoscopy completed within one year were included in this study. 120 patients (23%) had a proximal polyp detected at colonoscopy. The prevalence of proximal polyps and histologically advanced polyps was related to the size, number, and histology of the distal index polyp found at sigmoidoscopy. However, the absolute difference in prevalence of proximal polyps stratified by dings at sigmoidoscopy was small. A total of 5 adenocarcinomas were detected in the proximal colon. All proximal cancers detected at colonoscopy occurred in patients with a distal polyp less than 10 mm. Our data emphasize the importance of colonoscopy in all patients with a polyp detected at sigmoidoscopy independent of its size and histology. PMID- 11855547 TI - Colorectal cancer risk: the impact of evidence of a field effect of carcinogenesis on blinded diagnosis using an anti-adenoma antibody test performed on colonoscopic effluent. AB - To better define high-risk populations for colorectal cancer screening, we used anti-adenoma antibody Adnab-9, comparing colonic effluent and tissue field effects from 45 high-risk and 11 control patients. We included Adnab-9 binding at the tissue level to elucidate the impact of a field effect of carcinogenesis contributing to the outcome of the effluent binding test. In high-risk patients, 64% of the left-sided effluent samples were positive (P < 0.002); 67% showed a field effect (P < 0.006); 82% of combined tests were positive (P < 0.001), as compared to 9%, 18%, and 27% respectively, of controls. Adnab-9 binding correlates with increased colorectal cancer risk associated with a field effect of carcinogenesis. PMID- 11855548 TI - Prognostic value of expression of sialosyl-Tn antigen in colorectal carcinoma and transitional mucosa. AB - We examined the immunohistochemical expression of sialosyl-Tn antigen in the colorectal cancer tissues of 116 patients who underwent curative resection to determine the association between patient prognosis and the expression of sialosyl-Tn in two different tissues: carcinoma and transitional mucosa. Negative or positive expression of sialosyl-Tn in the carcinoma and transitional mucosal tissues were denoted as CA- or CA+ and TM- or TM+, respectively. Patients were classified into one of four groups: CA-/TM-, CA-/TM+, CA+/TM-, and CA+/TM+. CA /TM- was observed in 13 patients and CA-/TM+, CA+/TM-, or CA+/TM+ was observed in 103 patients. The difference in five-year survival between the CA-/TM- group and all the other groups was statistically significant (P = 0.0457), and multivariate analysis showed that sialosyl-Tn expression in at least one of the two tissues was an independent prognostic factor. We conclude that the evaluation of sialosyl Tn expression in carcinoma as well as transitional mucosa is useful for predicting survival in colorectal cancer patients. PMID- 11855549 TI - Neoplastic changes in saccharide sequence of dermatan sulfate chains derived from human colon cancer. AB - Decorin, a small proteoglycan containing a dermatan sulfate (DS) chain, is expressed abnormally in human colon cancer stroma. The aim of this study was to determine neoplastic changes in DS chains from human colon cancer and normal colonic mucosa. Proteoglycans were extracted from human colon cancer and normal colonic mucosa and successively digested with enzymes. The glycosaminoglycan obtained was fluoro-labeled with 2-aminopyridine at reducing terminals and fractionated by HPLC. Fluoro-labeled DS chains were collected and digested with bovine testicular hyaluronidase, followed by HPLC. The repeating disaccharide connected to the linkage region [glucuronosyl-galactosyl-galactosyl-xylosyl(2 aminopyridine)] of pyridylaminated DS chains from both types of tissue was glucuronosyl-N-acetylgalactosamine. The other glucuronic acid of the pyridylaminated DS chain was located 12 saccharides from the reducing terminal in colon cancer, and 18 saccharides from the reducing terminal in normal colon. The saccharide sequence of DS chains from human colon cancer is altered from that in normal colon. PMID- 11855550 TI - Bile duct infarction following intraarterial hepatic chemotherapy mimicking multiple liver metastasis: report of a case and review of the literature. PMID- 11855551 TI - Types of adrenoreceptors mediating responses of rabbit gastric muscularis mucosae. AB - This study investigated adrenoreceptor-mediated responses of muscularis mucosae from the fundic and antral ends of the rabbit gastric corpus. Norepinephrine induced fundic muscularis mucosae contractions were enhanced by propranolol and converted to relaxations by phentolamine. Methoxamine, but not clonidine, elicited large fundic contractions. Fundic muscle responded to low isoproterenol concentrations with atenolol- and butoxamine-resistant relaxations, and to high concentrations with atenolol-sensitive contractions. Norepinephrine evoked propranolol-resistant relaxations of antral muscularis mucosae that were enhanced by phentolamine. Methoxamine and clonidine elicited small antral contractions. Lower concentrations of isoproterenol caused atenolol-resistant antral relaxations that were enhanced by butoxamine; higher concentrations produced weak excitation. Fundic and antral relaxations to isoproterenol were abolished by cyanopindolol. Fundic muscularis mucosae possesses excitatory alpha1-, beta1- and inhibitory beta3-adrenoreceptors. Excitatory beta2- and inhibitory beta3 adrenoreceptors predominate in the antral region. The heterogeneous adrenoreceptor-mediated responses of the gastric muscularis mucosae suggest that adrenergic modulation of its motor activity is unlikely to be linked to acid secretion. PMID- 11855552 TI - Conjunctival edema: a marker of increased mortality in patients with advanced hepatic encephalopathy and hepatocellular failure. AB - Conjunctival edema in the absence of overt ophthalmologic disease is seen in some patients with advanced hepatic encephalopathy and hepatocellular failure. We conducted an observational study in an inner city teaching hospital. Three hundred patients with advanced hepatocellular disease were studied. Three hundred ten intensive care patients were used as controls. The appearance of conjunctival edema and its relation to mortality was noted. Conjunctival edema was seen in 50 patients and was associated with high mortality; 80% in our patients. In conclusion, there was a striking correlation between conjunctival edema and increased mortality in our study population. PMID- 11855553 TI - Effects of microwave irradiation on rat hepatic tissue evaluated by enzyme histochemistry for acid phosphatase. AB - Microwave coagulation therapy (MCT) has been applied to small hepatic carcinomas. To clarify the sequential changes in histology and the viability of the microwave irradiated tissue, we examined irradiated normal rat liver with enzyme histochemistry for acid phosphatase (AcP). In the samples immediately after irradiation, the margin of the irradiated region was indistinct in H&E stain, while AcP enzyme histochemistry disclosed well-demarcated distinct zones: an inner zone adjacent to the electrode without AcP activity and a surrounding outer zone with attenuated enzyme activity. In the inner zone, the nuclear staining with hematoxylin persisted for at least one month, whereas that in the outer zone disappeared 24 hr after irradiation and was accompanied by neutrophilic infiltration and then replaced by granulation tissue. Our results indicated that microwave irradiation caused tissue fixation in the inner zone and coagulative necrosis in the outer zone. Because microwave-fixed cells retained their morphology well, they appeared very similar to normal cells in H&E-stained section. Enzyme histochemistry may be useful for assessment of cellular viability after microwave irradiation, by enabling us to distinguish fixed cells from viable cells. PMID- 11855554 TI - Helicobacter pylori seroprevalence in patients with autoimmune hepatitis. AB - Autoimmune hepatitis is characterized by a continuing hepatocyte necrosis that usually progresses to liver cirrhosis. Autoimmunity is also a feature of chronic infection by Helicobacter pylori, a gram-negative bacterium involved in the pathogenesis of peptic ulcer and upper gastrointestinal bleeding, with both events frequently occurring in patients with chronic liver disease. A newly described pathogenetic mechanism for chronic hepatitis and hepatocellular carcinoma in the mouse is linked to Helicobacter spp. infection. A high prevalence of H. pylori infection was demonstrated in patients with viral-related cirrhosis but never studied in cases of autoimmune hepatitis. In a case-control study, we examined 31 consecutive patients (25 women and 6 men, age range 20-66, mean age 46 +/- 4.3 years) suffering from autoimmune hepatitis and 62 sex- and age-matched blood donors (50 women, 12 men, age range 20-65, mean age 46 +/- 5.4 years) resident in the same area. Antibodies to H. pylori were present in 20 of 31 (64.5%) autoimmune patients compared to 33 of 62 (53.2%) controls (P = 0.3, odds ratio 1.60, 95% CI 0.60-4.28). The difference was not statistically significant either in female or male patients. In conclusion, the prevalence of H. pylori infection in patients and controls was similar in our study of patients with chronic autoimmune hepatitis. PMID- 11855555 TI - Implementation of vaccination in patients with cirrhosis. AB - Vaccination for hepatitis A, hepatitis B, pneumococcus, and influenza in patients with chronic liver disease is recommended. Our purpose was to determine the degree of adherence to these recommendations in cirrhotics presenting for liver transplantation evaluation. Our sample consisted of 105 patients presenting for evaluation for liver transplantation. Data were obtained by medical record review and patient interview. The age of the patients was 52 +/- 9 years (mean +/- SD) and the Child-Pugh score was 9 +/- 2. Twenty-nine patients had natural immunity for hepatitis A. Of the remaining 76, 20 (26.3%) received vaccine. Of the 89 patients without evidence of prior exposure to hepatitis B, 23 (25.8%) received vaccine. Pneumococcal and influenza vaccination was performed in 36 (34%) and 58 (55%) respectively. In conclusion, vaccination for viral hepatitis, pneumococcus, and influenza not being consistently performed. Public health efforts aimed at raising awareness about implementing vaccination in these patients are strongly encouraged. PMID- 11855556 TI - Complications of partial splenic embolization in cirrhotic patients. AB - In recent years, partial splenic embolization (PSE) has been widely used in patients with cirrhosis and hypersplenism caused by portal hypertension. We investigated the complications associated with PSE cases seen in our hospital. Seventeen cases of liver cirrhosis that had undergone PSE were examined to investigate the complications associated with it. Mean infarcted area of the spleen was 66.2%. Leukocyte and platelet counts in 16 of 17 patients were seen to improve after PSE and persisted for at least one year. The most frequent side effects were abdominal pain (82.4%) and fever (94.1%). Severe side effects were seen in two of those 17 patients. One patient died from acute on chronic liver failure. The other patients contracted bacterial peritonitis and splenic abscess and needed drainage of splenic abscess before recovery. These two cases were in Child-Pugh class B. In conclusions, PSE is a useful treatment for patients with cirrhosis and hypersplenism caused by portal hypertension. However, the possibility of severe complications, especially in patients with noncompensated cirrhosis, should be kept in mind. PMID- 11855557 TI - Dopaminergic control of renal tubular function in patients with compensated cirrhosis. AB - In normal humans, plasma dopamine levels rise during head-out water immersion or saline intravenous infusion. Dopamine inhibits Na+,K+-ATPase activity in the proximal tubule and blunts aldosterone secretion leading to increased diuresis and natriuresis. The aim of this study is to evaluate the role of endogenous dopaminergic activity in the intrarenal sodium handling in patients with compensated liver cirrhosis. We studied nine healthy controls and 12 patients with Child-Pugh A cirrhosis during a normosodic diet for (1) dopaminergic activity, as measured by the incremental aldosterone responses 30 and 60 min after intravenous metoclopramide administration; (2) basal plasma levels of active renin and aldosterone; (3) 4-hr renal clearance of lithium (an index of fluid delivery to the distal tubule), creatinine, sodium, and potassium, first without and then with dopaminergic blockade with intravenous metoclopramide. The patients displayed greater endogenous dopaminergic activity, evidenced by higher incremental aldosterone responses compared with controls (+30 min: 160.2 +/- 68.8 vs 83.6 +/- 35.2 pg/ml, P < 0.01; +60 min: 140.5 +/- 80.3 vs 36.8 +/- 39.1 pg/ml, P < 0.01, respectively). In spite of this, patients and controls did not show significantly different basal aldosterone plasma levels, delivery of sodium to the distal nephron, or urinary excretion of sodium. In both groups the dopaminergic blockade with metoclopramide determined no change in sodium and potassium urinary excretion, but it caused a fall of the fluid and sodium delivery from the proximal tubule to the distal nephron among the patients (from 30.7 +/- 9.3 to 14.4 +/- 4.5 ml/min, P < 0.001; and from 4.25 +/- 1.30 to 2.00 +/ 0.64 meq/min, P < 0.001, respectively). In this group the natriuresis was maintained due to a reduction of the reabsorbed fraction of the distal sodium delivery (from 97.5 +/- 1.9% to 89.8 +/- 12.4%, P < 0.05). In conclusions, compensated cirrhotic patients display an increased endogenous dopaminergic activity compared with controls. This function is critical in maintaining the delivery of sodium to the distal nephron. PMID- 11855558 TI - Efficacy of irbesartan, a receptor selective antagonist of angiotensin II, in reducing portal hypertension. AB - The use of angiotensin II antagonists in the treatment of portal hypertension remains controversial. Our aims were to assess the effect of Irbesartan on portal pressure and to evaluate its safety in cirrhotic patients with portal hypertension. Twenty-five cirrhotic patients were treated in a pilot study with Irbesartan 300 mg orally once daily for 60 days. Hemodynamic evaluations and biochemical tests were performed before therapy and after two months of treatment. Three patients (12%) discontinued treatment for symptomatic arterial hypotension (mean arterial pressure -26.% +/- 3.1 versus basal). In the 18 responders, the hepatic venous pressure gradient diminished by a mean of 18.1% +/ 10.5 from baseline (p = 0.02); the gradient decreased by 20% or more in only 5 patients (23%). The mean arterial pressure decreased significantly during therapy (92 +/- 7 vs 109 +/- 25 mm Hg, P < 0.001). In conclusions, Irbesartan induced a marginal reduction in portal pressure and its safety was limited by the pronounced effects on arterial pressure. PMID- 11855559 TI - Long-term clinical remission induced by corticosteroid withdrawal therapy (CSWT) in patients with chronic hepatitis B infection: a prospective randomized controlled trial--CSWT with and without follow-up interferon-alpha therapy. AB - The effectiveness of corticosteroid withdrawal therapy (CSWT), with or without follow-up interferon-alpha (IFN-alpha), has not been reported for HBe antigen (HBeAg) -positive patients with chronic hepatitis B. We conducted a prospective randomized controlled trial in 42 patients with HBeAg- and HBV-DNA-positive chronic hepatitis B (HBV genotype C: 38 patients) to assess the possible additive effect of follow-up IFN-alpha after CSWT compared with CSWT alone. HBeAg seroconversion rates in the CSWT-alone and the combination group were 11.1% vs 11.8% at 24 weeks, 27.8% vs 12.5% at 52 weeks, 33.3% vs 18.8% at 76 weeks, and 38.9% vs 18.8% at 104 weeks, respectively. The final HBeAg seroconversion rates after CSWT alone were twice those following combination therapy. We conclude that CSWT alone is a very short-term treatment of just three weeks that may be more effective for long-term clinical remission than CSWT followed by IFN-alpha in Japanese genotype C-dominant hepatitis B patients. PMID- 11855560 TI - Reappearance of hepatitis B surface antigen in immunocompromised individuals: reinfection or reactivation? PMID- 11855561 TI - Porphyria cutanea tarda: multiplicity of risk factors including HFE mutations, hepatitis C, and inherited uroporphyrinogen decarboxylase deficiency. AB - The coexistence of factors considered to contribute to development of porphyria cutanea tarda was studied in 39 consecutive patients. Highly prevalent factors were alcohol intake in 79%, smoking in 86%, hepatitis C virus infection in 74%, estrogen use in 73% of 11 females, and at least one mutation in the HFE (hereditary hemochromatosis) gene in 65%. The C282Y mutation was found in 29%, H63D in 47%, and S65C in 0%. HFE genotypes included C282Y/C282Y in 9%, H63D/H63D in 9%, C282Y/H63D in 12%, C282Y/wild type in 9%, and H63D/wild type in 26%. Less prevalent were HIV infection in 15% (or 25% of those tested, N = 24) and erythrocyte uroporphyrinogen decarboxylase deficiency, which distinguishes familial (type 2) from "sporadic" (type 1) porphyria cutanea tarda, in 19%. Multiple contributing factors coexisted in both types 1 and 2, with 92% of all patients having three or more factors. These observations indicate that this porphyria is multifactorial in the individual patient, and therefore is seldom attributable to a single identifiable cause. Profiling for all potentially contributing factors is important for individualizing management. PMID- 11855562 TI - Human leukocyte interferon-alpha treatment for chronic HCV-related hepatitis in hemophilic patients previously intolerant to other interferons-alpha. AB - Thirty patients affected by hemophilia A or B or von-Willebrand's disease and chronic posttransfusional active HCV hepatitis who developed major side effects during the course of a previous treatment with recombinant interferon-alpha (IFN alpha) were studied. In all patients IFN-alpha therapy had to be discontinued and those who achieved a primary serologic and viral response to HCV relapsed within a few months. After a washout period, patients were retreated with human leukocyte IFN-alpha, 6 MU thrice weekly for 12 months. In about 90% of patients, a primary response, with normal AST and GGT values and undetectable HCV-RNA, was achieved within the third month of treatment and for the entire duration of treatment none of the patients had to discontinue therapy because of severe adverse reactions. During posttherapy follow-up only one patient relapsed. The human leukocyte IFN-alpha regimen looks to be very effective and safe for carriers of inherited clotting disorders who developed major side effects with recombinant IFN-alpha therapy for HCV-related chronic hepatitis. PMID- 11855563 TI - Prevalence and significance of hepatitis GB virus-c/hepatitis G virus viremia in a large cohort of patients with chronic hepatitis B infection, with chronic hepatitis C infection, and on renal replacement therapy in Hong Kong. AB - A total of 455 patients were recruited to study the prevalence of hepatitis GB virus-C/hepatitis G virus viremia in Hong Kong. There was no significant increase in the prevalence of hepatitis GB virus-C viremia in asymptomatic hepatitis B virus- and hepatitis C virus-infected patients compared to that of controls (1.56% and 7.14%, respectively, vs 3.85%, both P = NS). Renal patients as a whole had a significantly higher prevalence of hepatitis GB virus-C viremia compared to that of controls (13.95% vs 3.85%, P = 0.0271). The duration of the replacement therapy, especially for patients with peritoneal dialysis was associated with a higher chance of hepatitis GB virus-C viremia. Among renal patients, renal transplanted patients had the highest prevalence of hepatitis GB virus-C viraemia (19.1%) probably because of a higher susceptibility as a result of immunosuppression. However, hepatitis GB virus-C viraemia did not cause liver biochemistry derangement in renal transplanted patients. PMID- 11855564 TI - Amantadine's viral kinetics in chronic hepatitis C infection. AB - Treatment for hepatitis C virus infection is limited in patients not responding to traditional therapy. Amantadine is effective for influenza infections and has been studied in patients with hepatitis C. Our aim was to determine the efficacy and safety of amantadine and to study the viral kinetics following amantadine therapy. Twelve patients with detectable HCV antibodies received amantadine 100 mg, orally twice daily. Serial HCV RNA and ALT blood samples were drawn and adverse effects were evaluated. Mean HCV RNA levels did not decrease in the first 24 hr of amantadine therapy but did decline significantly by day 3, only to rebound by day 7. All HCV RNA levels remained detectable throughout therapy and were not different from baseline values. Thirty-three percent of the patients obtained normal ALT levels after the first 24 hr of treatment and levels remained within normal range throughout the study period. More than a third of the patients discontinued therapy due to severe adverse effects occurring within one to three months after initiating treatment. In conclusion, although amantadine therapy alone was not effective, it should be considered as an adjunctive form of therapy along with interferon and ribavirin. PMID- 11855565 TI - Nodular regenerative hyperplasia of the liver and focal global glomerulosclerosis associated with sickle cell anemia. PMID- 11855566 TI - Rapid temporary portosystemic bypass. AB - Extracorporeal venovenous bypass has been almost eliminated by the use of piggyback orthotopic liver transplantation. In some instances, a temporary internal portocaval shunt is constructed to relieve hemodynamic instability. We describe a quick and simple technique in which a short plastic cannula is used to bridge the distance between portal vein and IVC. PMID- 11855567 TI - Outcome of orthotopic liver transplantation in patients with hepatitis C. AB - Recurrence of chronic hepatitis C (HCV) after orthotopic liver transplantation (OLT) is universal. The published studies suggest that the short-term outcome is good in these patients, but the long-term prognosis remains unclear. The purpose of this study was to evaluate the outcome of patients with HCV undergoing OLT in a single center and to analyze the risk factors associated with poor outcome. In this retrospective study, we evaluated the outcome of 58 OLT patients with proven HCV who underwent OLT between February 1990 and April 1997 at our institution. The median follow-up time was 36.9 months. Recurrent posttransplant HCV hepatitis was confirmed by liver biochemistry, histology, and persistent HCV RNA in the serum. The patient and graft survival of patients with HCV was compared to that of 42 primary biliary cirrhosis (PBC) and 41 primary sclerosing cholangitis (PSC) patients transplanted during the same period. Following OLT, biochemical evidence of recurrent HCV hepatitis was absent in 46%. Forty percent of patients had recurrent HCV hepatitis and 14% had clinical evidence of recurrent HCV. Thirty one patients were on cyclosporine, 22 patients on tacrolimus, and 5 patients had cyclosporine switched to tacrolimus or vice versa. The recurrence rate of HCV chronic hepatitis was similar in patients who had cyclosporine (35.5%) or tacrolimus (45.5%) based immunosuppression. Eleven patients (19%) died and five patients (8.6%) were retransplanted for chronic rejection (two), mismatch (one), or primary graft nonfunction (two). The cumulative patient survival rates of one, three, and five years were 94.8%, 84.1%, and 62.2%, respectively. The severity of liver disease progressed with time; 8% of patients developed cirrhosis within two years. The survival rate did not show any relation between HCV recurrence and the type of immunosuppression. In conclusion, although the survival of patients with HCV was not statistically significant compared to those with PBC or PSC, there was a trend towards a lower five-year survival in HCV. PMID- 11855568 TI - Abdominal wall endometriomas. AB - Endometriosis is a condition in which uterine mucosal tissue is located outside the uterus. Endometriosis may be pelvic or extrapelvic. The term endometrioma is used when endometriosis appears as a circumscribed mass. Abdominal wall endometriomas are usually a secondary process in scars after surgical procedures. A retrospective study of abdominal wall endometrioma, from March 1992 through April 1999 at our institution was done. The mean age of the patients was 28.4 years. Twelve of these reported cases were secondary to previous surgery. One patient presented primarily with an abdominal wall mass without previous surgical history. The most common presentation was an abdominal wall mass associated with pain during the menstrual cycle. Endometrioma was considered as a differential diagnosis in seven patients. All patients underwent surgery. Along with the literature review on endometrioma, the importance of considering it in the differential diagnosis for patients of child-bearing age is discussed. PMID- 11855570 TI - Lack of evidence for fibrosing colonopathy by 5-ASA in humans. PMID- 11855569 TI - Interaction of fibronectin and aggregation substance promotes adherence of Enterococcus faecalis to human colon. AB - This in vitro study investigates the interaction between aggregation substance (AS), a virulence factor of Enterococcus faecalis, and colonic mucosal fibronectin in normal colon and colon from patients with Crohn's disease. Fibronectin was found to be overexpressed in Crohn's disease compared to normal colon. Compared to E. faecalis OG1X:pAM944 (AS-negative), E. faecalis OG1X:pAM721 (expressing AS) showed a significantly enhanced adhesion to human colonic mucosa in normal colon and in colon from patients with Crohn's disease. Double-staining of fibronectin and AS-positive enterococci showed that colocalization of bacteria and fibronectin was significantly more frequent in Crohn's disease than in normal colon. Preincubation of bacteria with soluble fibronectin caused a significant reduction in the adherence to fibronectin. In conclusion, the interaction between AS and fibronectin plays is an important factor that mediates adhesion of Enterococcus faecalis to colonic mucosa. This might be one of the mechanisms responsible for bacterial translocation of Enterococcus faecalis. PMID- 11855571 TI - Some epidemiological aspects of needle stick injuries among the hospital health care workers: Eastern Province, Saudi Arabia. AB - This study was conducted to provide some epidemiological aspects of needle stick injuries among health care workers in the eastern province in Saudi Arabia. Data about 282 injuries reported from the hospitals of the Eastern Province of Saudi Arabia were collected through specially designed questionnaire retrospectively from 1995 to 1997. Results showed that 50% of injuries occurred in the first 3 years of employment. Workers in medical and surgical specialties suffered an equal degree of reported exposure. The highest percentage (46.8%) of injuries occurred during syringe related actions. Patients' wards were the major location of incidents (48.5%) followed by ICUs and dialysis units (17.7%), theatres (15.6%) and accidents and emergency departments (13.8%). There was no significant association between injuries and type of shifts. The middle hours of the shifts showed a significant association with injuries. The pattern of injuries was significantly associated with over loaded periods of medical practices. Deficiencies in implementing the standard recommendations for prevention and control of needle stick injuries were noted in the studied hospitals. PMID- 11855572 TI - European origin of patients with Paget's disease of bone in the Buenos Aires area. AB - Paget's bone disease is heterogeneously distributed and several foci of high prevalence have been reported in Europe, United States, Argentina and Australia. The aim of the present work was to determine the ethnic origin of the disease in Buenos Aires using a cross sectional epidemiological study. Sample choice was based on a sampling according to grandparents' nationality. Ninety five percent of Paget patients were of European descent and 5% were non-European, while in the control group the proportion of European descendants is lower: 83% (OR: 3.7; p < 0.007; IC 95%: 1.4-9.7). Within the group of patients with Paget's disease the proportion of Italian and Russian descendants was higher than expected according to the 1914 Argentinean census. The prevalence of Paget's disease among European migrants was higher than in the control group of citizens. Regardless of environmental factors, it is likely that the migrants carried a higher risk of developing the disease. PMID- 11855573 TI - The completeness of brain tumour registration in Devon and Cornwall. AB - OBJECTIVES: To determine ascertainment rates for primary brain tumours and examine factors influencing those rates. DESIGN: Comparison of a clinical database with official figures from the Regional Cancer Intelligence Unit. RESULTS: Of 1480 potential cases 52% appeared in the registry. Only two-thirds of patients operated upon were registered and less than one-third of those who were not operated appeared in the registry. Independent predictors of registration were having had an operation, 5.47 (4.19-7.17) (odds ratio (95% CI)), being over 60 years of age, 1.62 (1.26-2.07), and having a tumour requiring radiotherapy, 2.52 (1.74-3.66). CONCLUSIONS: There is asymmetric registration of primary brain tumours in Devon and Cornwall with under registration of more benign tumours in younger patients. If similar problems exist across the United Kingdom, this could have a negative influence on survival rates, and international comparisons of such rates should be performed with caution. PMID- 11855574 TI - A comparison of smoking habits, beliefs and attitudes among Tuscan student nurses in 1992 and 1999. AB - Two-hundred-and-fifty students (aged 24+/-4 years, mean +/- SD, females were 75%) attending the first year of training in Florence Nursing School in 1991-1992 and 205 ones (aged 22+/-4 years; females 83% of the total) in the first year course of Nursing Schools of the Universities of Siena and Florence in 1998-1999 answered the same anonymous, self-administered questionnaire about their smoking habits, beliefs and attitudes. The overall response rate was 92% in 1992 and 88% in 1999. In 1992 and 1999, respectively, exsmokers were 13 and 11%, current smokers 51 and 43%. Ten percent of nurses who smoked in 1992 and 16% in 1999 were occasional smokers. The average daily number of cigarettes among respondents who smoked was lower in 1999. For both the surveys the beliefs about tobacco smoking remained generic and more often drawn from unspecific sources of information. Compared to 1992, in 1999 student nurses improved some attitudes towards smoking, such as the increased awareness that smoking cessation methods may be useful to quit and that hospital should be smoke-free. These positive attitudes did not change their smoking habits and the opinion on their ongoing smoking habits. Student nurses also did not obtain a consistently increased awareness about their role for advising subjects who smoked against smoking. We conclude that from 1992 to 1999 in Tuscany the smoking habits of first year student nurses changed little and remained high. There is a need for targeting antismoking education and for including specific training about tobacco smoking in the curricula of Italian nurse schools. PMID- 11855575 TI - Impact of a 10-year nation-wide alcohol campaign on knowledge of sensible drinking limits in Denmark. AB - In Great Britain and in Denmark, strong efforts have been made to influence knowledge on upper threshold for hazardous drinking. In Denmark, a campaign has been repeated every week 40 from 1990 to 2000 with information on the sensible drinking limits of 21 drinks per week for men and 14 drinks per week for women. The aim of this study was to examine the effect of this ongoing campaign on the level of knowledge of sensible drinking limits for men and women. Random representative samples of 1030 adult Danes were telephone interviewed each year during 1994-1999. Our main finding was that the level of knowledge of sensible drinking limits for own sex increased in all subsets of the population throughout the period. However, at the end of the study period (1999) a total of 80% of highly educated young (18-25 years) men knew sensible drinking limits for own sex, while only 35% of uneducated older (more than 65 years) men had knowledge on sensible drinking limits. The proportions were similar among women. Subjects admitting an intake higher than sensible for own sex, i.e. 21 and 14 drinks per week, respectively, had the highest knowledge of these drinking limits. We conclude that public health campaigns, such as the sensible drinking limit campaign, certainly has an impact on level of awareness in the general population. Furthermore, those drinking more than 21 and 14 drinks per week, respectively, are reached by these campaigns. PMID- 11855576 TI - Prevalence of markers for hepatitis A, B and C in the German population. Results of the German National Health Interview and Examination Survey 1998. AB - The prevalence of serological parameters indicative of infection with hepatitis A, B and C was determined using sera collected from representative population samples in the former East German (new) federal states and the West German (old) federal states during the German National Health and Examination Survey in 1998. Sera were tested for antibodies to hepatitis A virus (HAV), to hepatitis B core antigen (HBc) and to hepatitis B surface antigen (HBsAg), hepatitis C Virus (HCV), as well as for the presence of HBsAg and HCV-RNA. The mean weighted prevalence of anti-HAV was 46.5% (95% CI: 45.3-47.7) and increased markedly with age. The mean weighted prevalence of past infection with hepatitis B was 7.7% (95% CI: 7.0-8.4) in the old federal states and 4.3% (95% CI: 3.2-5.3) in the new federal states, corresponding to an overall prevalence of 7.0% (95% CI: 6.4-7.6). The mean weighted prevalence of HBsAg carriage was 0.6% (95% CI: 0.4-0.8), while the prevalence of HCV antibodies was 0.4% (95% CI: 0.2-0.5). PMID- 11855577 TI - Traumatic brain injury in Denmark 1979-1996. A national study of incidence and mortality. AB - In order to describe and analyse the development of the incidence of traumatic brain injury (TBI) in Denmark for different age groups of the two genders from 1979 through 1993 (for fatal injuries through 1996), a computerised search corresponding to diagnoses ICD 8th ed., 800, 801, 803, 850-854 from 1979 through 1993 was carried through in the national hospital register. Each person was counted only once, according to the most serious injury during the study period. For fatal cases, the search was extended till 1996. From 1979-1981 to 1991-1993, the total age-adjusted incidence of persons hospitalised under diagnoses ICD 800, 801, 803, 850-854 decreased 41% from 265 to 157 per 100,000 of the population per year. Decreases were 42% for ICD 850, brain concussion, 56% for ICD 800, 801, 803, cranial fractures, and 16% for ICD 851 854, structural brain injury. The percentage of cases with ICD 851-854 increased from 8.4 to 11.7% of the total. From 1979-1981 to 1985-1987 there was a 2% decrease in fatal TBI in and outside hospital (from 14.68 to 14.35 per 100,000), against a total 27% decrease to 10.67 in 1994-1996. For diagnoses ICD 851-854 and for fatal cases, significantly accelerating decreases from 1985-1987 were found only for the younger age groups. Consequently, in the period from 1979 to 1993, the mean age at injury increased by 10 years for persons hospitalised under diagnoses ICD 851-854. Decreases may be explained partly by changing admission and other hospital practices, and partly by the effect of comprehensive national preventive programs launched at the middle of the study period, the effect of which seemed to vary by age group and gender. PMID- 11855578 TI - Association between gastric cancer mortality and nitrate content of drinking water: ecological study on small area inequalities. AB - The carcinogenic feature of N-nitroso compounds has been well established. Similarly, the transformation of ingested nitrate to N-nitroso compounds in the stomach has been thoroughly documented, nevertheless nitrates' carcinogenic effect has not been proved convincingly in human. The present study was aimed to investigate a population of small villages provided by drinking water with high and widely variable nitrate content (72 mg/l median, 290.7 mg/l 95-percentile concentration). Empirical Bayes estimates for settlement-specific age-, sex-, and year-standardised mortality ratios of gastric cancer (GC) were related to the settlement level average nitrate concentrations in drinking water controlling for confounding effects of smoking, ethnicity and education. The log-transformed average nitrate concentration showed significant positive association with stomach cancer mortality in linear regression analysis (p = 0.014). The settlements were aggregated according to the nitrate concentration into 10 percentile groups and the standardised mortality ratios (SMRs) were calculated. Those groups with higher than 88 mg/l average nitrate concentration showed substantial risk elevation and the log-transformed exposure variables proved to be significant predictors of mortality (p = 0.032) at this level of aggregation also. The association seemed to be fairly strong (r2 = 0.46). Although this investigation constituting an ecological study has certain limitations, it supports the hypothesis that the high level of nitrate in drinking water is involved in the development of GC. PMID- 11855580 TI - Absence of an association between serum uric acid and mortality from cardiovascular disease: NIPPON DATA 80, 1980-1994. National Integrated Projects for Prospective Observation of Non-communicable Diseases and its Trend in the Aged. AB - Although elevated serum uric acid has been associated with an increased risk of cardiovascular disease, its importance as a risk factor is still controversial. The authors examined the relationship between serum uric acid levels and death from all causes, including cardiovascular disease and stroke. The baseline data were collected in the National Cardiovascular Survey in 1980. The survey was carried out for all household members aged 30 years or older in 300 districts, which were randomly selected throughout Japan. The number who participated in the survey was 10,897. The vital status was ascertained in 1994. Finally, 8172 subjects were available for the analyses. There were 108,284 person-years of follow-up, and 960 deaths from all causes, 249 deaths from cardiovascular disease, and 174 deaths from stroke. After adjustment for age and other cardiovascular disease risk factors, uric acid levels were not associated with mortality from all causes, cardiovascular disease, or stroke. These findings indicate that serum uric acid levels are not related to increased risk for death from all causes, including cardiovascular disease and stroke in a Japanese population. PMID- 11855579 TI - Socio-economic position and cardiovascular risk factors in an Italian rural population. AB - Cardiovascular disease (CVD) is the leading industrial world cause of mortality. Lower social class groups have higher incidence of CVD and also display less favourable risk factor profiles. To investigate the association and gradient between major cardiovascular risk factors (smoking habit, serum lipid profile, blood pressure, relative body weight) and socio-economic position (proxy measures selected: education and occupation) data on 2592 men and 2866 women were analysed. The effect of potential confounding factors and effect modifiers was estimated. Linear and logistic regression modelling was performed for continuous and dichotomous outcomes respectively. The lower the grade of employment or the level of education, the higher was the prevalence of obesity in the study population. The association was stronger in women than in men. Higher education was associated with a lower prevalence of smoking among men and a higher prevalence among women. Systolic blood pressure was negatively related to socio economic position in both men and women in the age-adjusted models. Attention should be concentrated on socio-economic differences in obesity and blood pressure in this population. PMID- 11855581 TI - Differences in all-cause, cardiovascular and cancer mortality between Hong Kong and Singapore: role of nutrition. AB - BACKGROUND: The majority of inhabitants in Hong Kong and Singapore are ethnic Chinese, but all-cause and cardiovascular mortality rates in these two regions are markedly different. This study describes differences in the magnitude and trends in mortality and attempts to explain these differences. METHODS: Data of mortality rates in 1963-1965 and 1993-1995 in the age class of 45-74 years, dietary habits and other factors were compared between Hong Kong and Singapore using Japan, Spain and the USA as reference countries. Mortality and food consumption data were obtained from WHO and FAO, respectively. RESULTS: Large differences in all-cause and cardiovascular mortality exist between Hong Kong and Singapore. The difference in total cancer mortality was less consistent and smaller. The most pronounced finding was that ischemic heart disease mortality in 1993-1995 was 2.98 and 3.14 times higher in Singapore than in Hong Kong in men and women, respectively. Of the five countries considered, Singapore has the highest all-cause mortality in both sexes in the period of 1960-1995. The ratio of animal to vegetal fat was higher in Singapore (2.24) than in Hong Kong (1.08). Singapore had higher serum concentrations of total cholesterol and low-density lipoprotein cholesterol than Hong Kong, but the opposite result was observed for high-density lipoprotein cholesterol. CONCLUSIONS: There are striking differences in all-cause and cardiovascular mortality between Hong Kong and Singapore. These differences can be most reasonably and plausibly explained by their differences in dietary habits, for example, a higher consumption of coconut and palm oil, mainly containing saturated fat, in Singapore. PMID- 11855582 TI - Anonymous reporting of HIV infection: an evaluation of the HIV/AIDS surveillance system in Norway 1983-2000. AB - Several European countries are considering implementing surveillance systems for HIV infection, but questions remain regarding patient confidentiality. The population-based HIV/AIDS surveillance system in Norway integrates anonymous HIV case reports from laboratories and clinicians and named AIDS case reports. This evaluation includes a description of the system, evidence of system attributes, estimation of resources for system operations, and documentation of the system's usefulness. HIV case reports provide a far better picture of the epidemic than AIDS reports. The median delay between positive HIV test and reporting was 30 days (interquartile range 18-49 days). Completeness of demographic and epidemiologic information in the surveillance database ranges from 60 to 100%. Information on pre-AIDS mortality and emigration is incomplete. The system cost euro 25,200 in 1999. Results are published every week and used for planning of health care and prevention. We conclude that the Norwegian surveillance system with anonymous reporting of HIV cases is simple, inexpensive and flexible, and capable of providing a representative and timely overview that guides prevention. The system fulfils its objectives while respecting confidentiality and adhering to ethical principles. A similar system may be considered in other countries. PMID- 11855583 TI - Hansenula anomala outbreak at a surgical intensive care unit: a search for risk factors. AB - During a 5-month period, Hansenula anomala (H. anomala), an opportunistic fungus, caused an outbreak of infections in eight adult patients treated at a surgical intensive care unit (ICU). The source of the infections and route of transmission could not be identified. A case-control study included 32 patients treated simultaneously at the surgical ICU. Univariate analysis pointed to the following significant risk factors: blood alkalosis, reduced urea, duration of hospitalization, bacteremia and colonization with Pseudomonas aeruginosa, and an APACHE II score >17 (during bacteremia or fungemia). The stepwise logistic regression multivariate analysis showed only the duration of blood alkalosis to be significant in case patients. PMID- 11855584 TI - 'CHIP'ping a way through the cancer maze. PMID- 11855585 TI - Medico-social dimensions of menopause: a cross-sectional study from rural south India. AB - BACKGROUND: The quality of life of the increasing ageing population is becoming an important issue in India. There are very little data on the effect of menopause on women, especially from rural India. METHOD: A population-based cross sectional study was conducted on perceptions regarding menopause, prevalence of menopausal symptoms and association of family environmental factors with menopausal symptoms among 100 postmenopausal and 100 premenopausal rural women in south India. RESULTS: Fifty-seven per cent of postmenopausal women perceived menopause as convenient. Sixty-nine per cent of them complained of diminishing abilities after menopause. Twenty-three per cent felt that sexual life ends with the onset of menopause. Sixteen per cent reported that their husbands had become disinterested in them after menopause and I1% were apprehensive about the loss of femininity. A higher proportion of postmenopausal women reported hot flushes, night sweats, urge incontinence and other somatic symptoms as compared to premenopausal women. Fifty-four per cent of postmenopausal and 32% of premenopausal women were currently not sexually active. Fifty-nine per cent of postmenopausal and 38% of premenopausal women expressed loss of sexual desire and this difference was statistically significant. There was no significant association between menopause and depression. A poor perceived relationship within the family was shown to have a significant association with depression. There was a significant association between multiple somatic symptoms and menopause. CONCLUSION: A significantly higher proportion of postmenopausal women suffer from vasomotor symptoms, urge incontinence, loss of sexual desire and multiple somatic symptoms. They do not link these symptoms with menopause. Poor family environmental factors have a stronger association with depression than menopause. In view of these findings, it is important to determine the feasibility and impact of hormone replacement therapy in preventive health care in rural India. PMID- 11855586 TI - Germ cell tumours of the testis: clinical features, treatment outcome and prognostic factors. AB - BACKGROUND: The prognosis of patients with germ cell tumours of the testis has Improved over the past two decades following cisplatinum-based chemotherapy. Currently, staging and risk assessment of the disease is crucial in order to provide curative therapy for patients with poor risk features and not over-treat good risk patients. METHODS: We reviewed the case records of 71 men diagnosed to have germ cell tumours between January 1993 and October 1999. Their clinical characteristics, staging, treatment outcome and prognostic factors for response and survival were analysed. RESULTS: The median age of the patients was 30 years (range: 3-65 years); 69% were in the third and fourth decades. Sixty-one patients (86%) had a primary testicular tumour while in 10 (14%) the tumour was extragonadal. Histopathologically, 53 patients (75%) had non-seminomatous germ cell tumours and 15 (21%) had a seminoma. Twenty-seven patients (62%) had evidence of metastatic disease at the time of diagnosis. On prognostication, non seminomatous germ cell tumour patients could be divded into good, intemediate and poor prognostic groups comprising 41%, 17% and 40% of patients, respectively. All patients with a seminoma were in the good prognostic subgroup. Fifty-eight patients were evaluable for response. Overall, 91% of patients responded: complete response 71% and partial response 20%. Complete response rates were signiflcantly higher for the good risk (95%) compared to the intermediate (49%) and poor risk (47%) categories (p< 0.003). At a median follow up of 26 months, the 2-year overall and progression-free survival for all patients was 70% and 57%, respectively. The predictors for decreased overall and progression-free survival were age >35 years, presence of poor risk features and mediastinal primary disease. CONCLUSION: The outcome for germ cell tumours in men with good risk is excellent. A protocol consisting of bleomycin, etoposide and cisplatin is effective. Tailoring of chemotherapy In good risk patients to minimize toxicity and Improving results in poor risk patients are areas that need further work. PMID- 11855587 TI - Outbreak of leptospirosis after the cyclone in Orissa. AB - BACKGROUND: Two weeks after the cyclone during October-November 1999, several persons in Orissa suffered from a febrile illness with haemorrhagic manifestations. Serum samples from a few such patients tested positive for anti leptospiral IgM antibodies. We conducted a study in four villages that were flooded after the cyclone to examine the possibility of leptospirosis being the cause of the outbreak. METHODS: One hundred forty-two persons living in four flooded villages in the Jaipur district of Orissa were interviewed for their disease history and possible risk factors after the cyclone. Blood samples were collected and tested for anti-leptospiral antibodies using the microscopic agglutination test, IgM ELISA and lepto-dipstick. Follow up samples were collected from those who had inconclusive results on the first test and the microscopic agglutination test was repeated on these samples. RESULTS: Eighty four of the 142 study subjects had suffered a febrile illness and 40 of them had positive results in one or more IgM-based tests and 28 had a positive microscopic agglutination test result as well. Thus, 19.2% of the study subjects (28/142) had serological evidence of symptomatic leptospiral infection after the cyclone. Also, 8.5% of the subjects had low levels of antibodies indicative of the level of background seroprevalence. CONCLUSIONS: The results indicate that there was an outbreak of leptospirosis in the flooded villages and the attack rate was high. A carrier state might have existed in the animal population and the cyclone and floods changed the environment drastically making it conducive for transmission of infection. Large numbers of persons were continuously exposed to flood waters and this resulted in the outbreak. PMID- 11855588 TI - Adverse drug reaction monitoring: knowledge, attitude and practices of medical students and prescribers. AB - BACKGROUND: Adverse drug reactions (ADRs) contribute to excessive health care costs through increased patient morbidity and mortality. Thus, there is an urgent need to create awareness among physicians towards ADR monitoring. The present study was designed to assess the knowledge, attitude and practices of fifth semester undergraduate students and prescribers (interns, junior residents and senior residents) towards the recording and reporting of ADRs. METHODS: The fifth semester MB,BS undergraduate students (n=107) and prescribers (n=l 17) working in different disciplines of Lady Hardinge Medical College and associated hospitals were given a questionnaire to answer. The responses of the undergraduate students were compared with those of prescribers. RESULTS: Knowledge about definition, classification, objectives and methods of ADR monitoring was found to be comparable in both groups. Spontaneous and intensive methods of ADR monitoring were known to the majority of participants of both groups. Attitude and practices of the prescribers were significantly (p<0.01 ) better with regard to the status of ADR monitoring in the institute. A significantly higher (p<0.001) proportion of prescribers (82%) as compared to the undergraduate students (64.5%), felt that ADRs should be reported both when it causes inconvenience to the patient as well as death. ADRs were encountered by both undergraduates (46%) and prescribers (66%) during their clinical project exercises and patient care, respectively. Commonly encountered ADRs were allergic reactions, symptoms of upper gastrointestinal irritation, extrapyramidal symptoms and hepatitis. The common offending groups of drugs causing these ADRs were non-steroidal anti-inflammatory drugs, antidopaminergics and chemotherapeutic agents. A majority of ADRs were suspected and subsided on their own by either stopping the drug or reducing its dose. CONCLUSIONS: The knowledge, attitude and practices of both undergraduates and prescribers were comparable but need further improvement. This suggests the need for suitable changes in the undergraduate teaching curriculum. Further, the prescribers also need periodic reinforcement regardingADR monitoring. PMID- 11855589 TI - Complications of endoscopic retrograde cholangiopancreatography and endoscopic sphincterotomy: diagnosis, management and prevention. AB - Endoscopic retrograde cholangiopancreatography and endoscopic sphincterotomy are carried out for a variety of billo-pancreatic diseases. These procedures were earlier performed at only a handful of tertiary centres in India. However, of late, they are being performed at several centres and even smaller clinics. Although, endoscopic sphincterotomy is one of the most commonly performed endoscopic procedure, there are no data on the number of procedures performed each year in India. It is estimated that in the USA alone more than 200,000 procedures are performed each year. The procedure has the dubious distinction of being considered as one of the most hazardous of all endoscopic procedures and is associated with a small but significant morbidity and mortality. The immediate complications of endoscopic sphincterotomy include pancreatitis, haemorrhage, perforation and cholangitis in addition to those associated with the use of sedation and anaesthesia. There are also reports of the long term adverse effects of the destruction of the sphincter of Oddi. There is also controversy as to what should be the minimum requirements for training and who should be allowed to perform these procedures. PMID- 11855590 TI - Ethics in surgical practice: an Indian viewpoint. PMID- 11855591 TI - Tackling hunger, disease and 'internal security': official medical administration in colonial eastern India during the Second World War (part I). PMID- 11855592 TI - Hard times. PMID- 11855593 TI - The jhoola bed. PMID- 11855594 TI - Candidiasis in HIV-infected patients: a clinical and microbiological study. PMID- 11855595 TI - Estimates of the years-of-life-lost due to the top nine causes of death in rural areas of major states in India in 1995. AB - BACKGROUND: Years-of-life-lost (YLL) contribute nearly two-thirds of the disability-adjusted life-years (DALYs) worldwide and are especially Important for India where infant and child mortality is still high. These were estimated for India under the Global Burden of Disease study for the year 1990. No estimates are available for the different states of India. We aimed to prepare state-wise estimates of YLL for different causes of death in rural areas and to determine the causes responsible for a higher burden in different states. METHOD: Percentage deaths of the top 9 causes reported in the Registrar-General's Survey of Causes of Deaths (Rural)--1995 in 13 major states of India and different age groups was applied to the expected number of total deaths. The life lost according to the standard life-table was age-weighted and discounted using the methodology of the Global Burden of Disease 1990 study. The causes of death were based on lay reporting which otherwise seem reliable. RESULTS: The all-cause YLL in rural India in 1995 were 207 per 1,000 population. The minimum was 74 in Kerala and maximum 276 in Madhya Pradesh. Pneumonia was the top cause responsible for 15 YLL. The inter-state variation was high as Tamil Nadu had only 1.6 and Uttar Pradesh 30.5 YLL from this cause. Cancers were a uniform burden across the states. Heart attack, and bronchitis and asthma cut across the more and less developed states. Suicides were a heavy burden in Andhra Pradesh and vehicular accidents in Haryana and Rajasthan. Bihar, Gujarat, Madhya Pradesh, Orissa, Rajasthan and Uttar Pradesh had communicable and nutritional conditions as predominant causes while Kerala and Punjab had non-communicable diseases as the predominant cause of YLL due to premature mortality. CONCLUSION: These results provide a new perspective about the causes of death that need more attention in rural areas of different states of India. These will also help prioritize areas which require more inputs at the state-level and hence will be useful for health policymakers. PMID- 11855596 TI - Reader survey on RN shortage: results. PMID- 11855598 TI - Pain management should be a priority. PMID- 11855597 TI - Use of nasal cannulae. PMID- 11855599 TI - Need for end-of-life care literature. PMID- 11855600 TI - Measuring end-tidal carbon dioxide: clinical applications and usefulness. PMID- 11855601 TI - Using high-frequency oscillatory ventilation to treat adults with acute respiratory distress syndrome. PMID- 11855603 TI - Using liquid ventilation to treat patients with acute respiratory distress syndrome: a guide to a breath of fresh liquid. AB - Research with liquid ventilation indicates that this technology may benefit patients with acute lung diseases such as ARDS. In addition to improving gas exchange, this novel therapy may enhance the delivery of antibiotics and other drugs and thus hasten the recovery from other serious diseases. Additional research on using perflubron in combination with other developing therapies is under way. Clearly, liquid ventilation is not a cure for ARDS. However, it may be helpful in the supportive management of patients with ARDS. As the phase 3 clinical trials are completed, liquid ventilation may become a part of treatment in the intensive care unit. Understanding how liquid ventilation works and knowing what to look for in patients treated with PLV will help ensure better outcomes for patients when this therapy becomes part of clinical practice. PMID- 11855602 TI - An easy method for interpreting the results of arterial blood gas analysis. AB - Interpretation of acid-base disturbances is an essential skill for critical care nurses. Using the H model makes this process easy. When students and novice critical care nurses feel competent with certain skills, their confidence levels are greatly enhanced. One of us (K.M.K.) has been using the H model for many years to teach students how to interpret the results of arterial blood gas analysis. The students are often amazed at how easy and fun the model makes learning a subject many perceive as complex. PMID- 11855604 TI - Pediatric critical care nursing: Annie's story. PMID- 11855605 TI - Correct procedures for performing atrial electroardiograms (ECGs) using the 5 lead bedside cable and epicardial pacing wires. PMID- 11855606 TI - Coronary care unit relies on nurses' feedback for recruitment and retention. PMID- 11855607 TI - Clinical judgment in evidence-based practice. PMID- 11855608 TI - Doctorally prepared nurses in clinical settings. PMID- 11855609 TI - Developing an evidence-based procedure: maintenance of central venous catheters. AB - This article describes the creation of an evidence-based guideline as part of a learning experience of a group of clinical nurse specialists. The guideline was the product of a utilization-focused integrated review of the literature conducted with the guidance of a nurse researcher. Key aspects of the evidence based recommendations, as well as factors that facilitated or hindered this effort, are shared. PMID- 11855610 TI - Controlled-release oxycodone hydrochloride (OxyContin). PMID- 11855611 TI - Health, humanities, and the arts. PMID- 11855612 TI - Malpractice target: can cardiac arrests really be prevented? PMID- 11855613 TI - Finding your way on the Internet. PMID- 11855614 TI - Care considerations for the client who is obese. PMID- 11855615 TI - Understanding the bariatric client and providing a safe hospital environment. AB - Mobility is a basic human need that, when not met, leads to a cascade of physical problems. Meeting the challenge of safe mobility for bariatric patients is a concern for many healthcare institutions. More than 20% of the adult population now has a body mass index equal to or greater than 30 and are classified as obese. Obesity is associated with many well-known health risks, including heart disease, metabolic disorders, and cancer. Nurses should be aware of these health risks, the interplay between mobility and health risks, and the national guidelines for treating the obese patient population. This article presents an overview of national guidelines for identifying, evaluating, and treating bariatric patients and identifies issues that create challenges to providing a safe environment for mobility among hospitalized obese patients. PMID- 11855616 TI - Understanding staff nurse responses to change: utilization of a grief-change framework to facilitate innovation. 1994 [classical article]. PMID- 11855617 TI - Protein quality of four types of edible mushrooms found in Jordan. AB - This study was conducted to determine the protein quality of four types of edible mushrooms, Terfezia claveryi, Pleurotus ostreatus, Tricholoma terreum and Agaricus macrosporus that are common in Jordan. Protein efficiency ratio (PER) and net protein utilization methods (NPU) were used. The four mushroom types failed to support growth as reflected by negative values for PER; -1.76 for T. claveryi; -0.23 for P. ostreatus; -0.98 for T. terreum and -0.41 for A. macrosporus. The NPU(op), NPU(st) and NdpE% for T. claveryi were 32.6, 33.9 and 3.89%, respectively. The respective values for P. ostreatus were 38.5, 40.7 and 4.35%; T. terreum 29.1, 29.8 and 3.63%; and A. macrosporus 31.5, 32.9 and 3.91%. The results showed that the four mushrooms were significantly (p < or = 0.05) lower in protein quality compared with casein results (NPU(op), NPU(st) and NDpE% of 78.5, 86.4 and 7.16%, respectively. PMID- 11855618 TI - Protein digestibility using corrected amino acid score method (PDCAAS) of four types of mushrooms grown in Jordan. AB - This study was conducted to evaluate the protein quality of four types of edible mushrooms common in Jordan in terms of protein digestibility-corrected amino acid score (PDCAAS), which is a combination of the chemical score of the limiting amino acid multiplied by true digestibility of the protein. True protein digestibility values were low (61.4, 73.4, 52.6 and 80.5 for Terfezia claveryi, Pleurotus ostreatus, Tricholoma terrum and Agaricus macrosporus, respectively). Based on the essential amino acids pattern requirements for children, the limiting amino acids in P. ostreatus and A. macrosporus protein were sulphur containing amino acids with chemical scores of 0.61 and 0.50, respectively. However, lysine was the limiting amino acid in the protein of T. claveryi and T. terreum with chemical scores of 0.71 and 0.67, respectively, and the PDCAAS was 0.43, 0.45, 0.35 and 0.40 for the mushrooms types, respectively. Considering the pattern of essential amino acid requirements of laboratory rats, the sulphur containing amino acids were the limiting amino acids in the protein of T. claveryi, P. ostreatus, T. terreum and A. macrosporus with chemical scores of 0.56, 0.30, 0.34 and 0.25, respectively. The PDCAAS were 0.34, 0.22. 0.17 and 0.20, respectively. It is concluded that the four mushroom types studied are of low protein quality. PMID- 11855619 TI - Flow behavior characteristics of ice cream mix made with buffalo milk and various stabilizers. AB - Ice cream made with buffalo milk, using optimum levels of various stabilizers of plant origin, was evaluated for its flow behavior characteristics, with the objective of producing an acceptable quality product. The minimum variation in the viscosity of mix was observed at three rates of shear (348.88, 523.33 and 1046.66 S(-1)) for all ice cream mixes. The flow behavior index (n) of all the mixes having optimum levels of various stabilizers was observed to be less than 1; indicating their pseudoplastic nature. Consistency coefficient (m) of sodium alginate was found to be 1.19; highest among all the stabilizers, followed by gelatin (1.17), karaya (1.08), guar gum (0.75), acacia gum (0.70), ghatti gum (0.36), and the control (0.29). The consistency coefficient (m) signifies the apparent viscosity of the pseudoplastic fluid. The viscosity of the mixes having various stabilizers (optimum levels) was found to be in descending order: Sodium alginate, gelatin, karaya, guar gum, acacia, ghatti and control. PMID- 11855620 TI - Efficacy of turmeric on blood sugar and polyol pathway in diabetic albino rats. AB - In the traditional system of medicine, Ayurveda, several spices and herbs are thought to possess medicinal properties. Among the spices, turmeric rhizomes (Curcuma longa. Linn.) are used as flavoring and coloring agents in the Indian diet everyday. In this research, we studied the effect of turmeric and its active principle, curcumin, on diabetes mellitus in a rat model. Alloxan was used to induce diabetes. Administration of turmeric or curcumin to diabetic rats reduced the blood sugar, Hb and glycosylated hemoglobin levels significantly. Turmeric and curcumin supplementation also reduced the oxidative stress encountered by the diabetic rats. This was demonstrated by the lower levels of TBARS (thiobarbituric acid reactive substances), which may have been due to the decreased influx of glucose into the polyol pathway leading to an increased NADPH/NADP ratio and elevated activity of the potent antioxdiant enzyme GPx. Moreover, the activity of SDH (sorbitol dehydrogenase), which catalyzes the conversion of sorbitol to fructose, was lowered significantly on treatment with turmeric or curcumin. These results also appeared to reveal that curcumin was more effective in attenuating diabetes mellitus related changes than turmeric. PMID- 11855621 TI - Vitamin C and quercetin modulate DNA-damaging effect of N-methyl-N'-nitro-N nitrosoguanidine (MNNG). AB - The effects of natural compounds, vitamin C and quercetin, present in fruits and vegetables, on the DNA damaging activity of a food carcinogen N-methyl-N'-nitro-N nitrosoguanidine (MNNG) were examined using the comet assay. Vitamin C, at a concentration of 50 microM, inhibited MNNG-induced DNA damage in human lymphocytes. Quercetin, up to a concentration of 10 microM, increased the extent of DNA damage, but at concentrations above 10 microM decreased damage below control values. Furthermore, quercetin had a strong antioxidant activity against oxidative damage evoked by H2O2 at 10 microM. The results obtained suggest that vitamin C and quercetin may have anti- or pro-oxidative properties depending on the state of the cell. PMID- 11855622 TI - Traditional production, consumption and storage of Kunu--a non alcoholic cereal beverage. AB - A survey of the production, consumption and storage of Kunu was carried out. Some of the information included consumption rate, processing techniques and equipment, producer's status and grains used. About 73% consume Kunu daily, 26% occasionally; 1% know it is produced but may or may not be consuming it. Millet (Pennisetum typhoideum), sorghum (Sorghum vulgare), maize (Zea mays), rice (Oryza sativa) and acha (Digitalis exilis) were used in its production in decreasing order of preference. The grains were used singly or combined; sorghum/millet was the most common combination in a ratio of 1:2 (w/w). Steeping was done in ordinary water for 12-72 h, depending on the grain type, in local clay pots, plastic buckets, calabashes or basins or 5-7 h in warm water (60-70 degrees C). The grains were dry or wet milled with or without spices such as ginger, red pepper, black pepper, clove and garlic. Other ingredients introduced included: sweet potatoes, malted rice, malted sorghum and Cadaba farinosa crude extract. Both dry and wet milling was done with grinding mills, mill stones or mortar and pestle, depending on locality. The product was then cooked into a thin free flowing gruel. The various types of kunu were: Kunun zaki, Kunun gyada, Kunun akamu, Kunun tsamiya, Kunun baule, Kunun jiko, Amshau and Kunun gayamba. Kunun zaki was the most commonly consumed. Production and consumption cut across all social classes of the society. PMID- 11855623 TI - Nutritional composition and antinutritional factors of chickpeas (Cicer arietinum L.) undergoing different cooking methods and germination. AB - The effects of cooking treatments (boiling, autoclaving and microwave cooking) and germination on the nutritional composition and antinutritional factors of chickpeas were studied. Cooking treatments and/or germination caused significant (p < 0.05) decreases in fat, total ash, carbohydrate fractions, antinutritional factors, minerals and B-vitamins. Germination was less effective than cooking treatments in reducing trypsin inhibitor, hemagglutinin activity, tannins and saponins; it was more effective in reducing phytic acid, stachyose and raffinose. Cooking treatments and germination decreased the concentrations of lysine, tryptophan, total aromatic and sulfur-containing amino acids. However, cooked and germinated chickpeas were still higher in lysine, isoleucine and total aromatic amino acid contents than the FAO/WHO reference. The losses in B-vitamins and minerals in chickpeas cooked by microwaving were smaller than in those cooked by boiling and autoclaving. Germination resulted in greater retention of all minerals and B-vitamins compared to cooking treatments. In vitro protein digestibility, protein efficiency ratio and essential amino acid index were improved by all treatments. The chemical score and limiting amino acid of chickpeas subjected to the various treatments varied considerably, depending on the type of treatment. Based on these results, microwave cooking appears to be the best alternative for legume preparation in households and restaurants. PMID- 11855624 TI - Localizing the movement proteins of Abutilon mosaic geminivirus in yeast by subcellular fractionation and freeze-fracture immuno-labelling. AB - The movement proteins BC1 and BV1 of Abutilon mosaic geminivirus fused to glutathion-S-transferase (GST) and Flag-peptide were expressed in fission yeast (Schizosaccharomyces pombe) cells to analyse the fundamental intracellular distribution of these proteins in an eukaryotic cell in the absence of plant specific factors. Most of BC1 protein sedimented rapidly after cell lysis and differential centrifugation. Using freeze-fracture immuno-labelling, the protein was detected in situ predominantly at plasma membranes and to a lower extent at cytoplasmic vesicles but not in the cytoplasm, the nuclei, or the mitochondria. Anti-BC1, anti-GST, and anti-Flag antibodies tagged smooth flecks only at the protoplasmic faces of the plasma membrane. The consequences of the BC1 behaviour for its use in two-hybrid analysis in yeast are discussed. In contrast, BV1 was detected mainly in the nucleus and partially in the cytoplasm but never associated with membranes. PMID- 11855625 TI - A pan-specific promoter activity of the 213bp segment of the pseudorabies virus early protein 0 gene in transgenic mice. AB - Pseudorabies virus (PrV) early protein 0 (EP0) is a transactivator that plays important roles in the viral gene expression. To examine a promoter regulatory element for the EP0 gene expression in vivo, we have generated two transgenic mouse lines expressing the EP0 gene under the control of a 213bp 5'-flanking sequence of the EP0 gene. To analyze the tissue specificity of transgene expression, mRNA of the EP0 gene was monitored in various tissues from the transgenic mice by the reverse transcriptase (RT)-polymerase chain reaction (PCR) analysis. The EP0 gene expression was demonstrated in all tissues tested by the RT-PCR analysis. These tissues included skin, muscles (skeletal and heart), lung, liver, spleen, small intestine, kidney, brain and testis. These results indicated that the 213bp sequence of the 5'-flanking region of the EP0 gene is capable of driving expression of the EP0 gene in vivo and the promoter is pan-specific. PMID- 11855626 TI - Baculovirus expression and antigenic characterization of the capsid proteins of three Norwalk-like viruses. AB - Human caliciviruses (HuCVs) are antigenically diverse. The antigenic relationships among different HuCVs have been difficult to study because HuCVs cannot be passaged in the laboratory. In this study, we describe cloning, sequencing and expression of the viral capsid proteins of three HuCVs that were identified in outbreaks of acute gastroenteritis in Virginia in 1997-1998. Yields of the capsid proteins similar to previously expressed recombinant Norwalk virus were obtained using the baculovirus expression system. Recombinant VA97207 capsid protein (rVA97207) and rVA98387, but not rVA98115, formed virus-like particles (VLPs). All three recombinant capsid antigens detected seroresponses in patients involved in outbreaks of acute gastroenteritis associated with genetically homologous or related HuCVs. The antigenic relationships of the three strains were further characterized using hyperimmune antisera against the three capsid antigens as well as four previously characterized recombinant capsid antigens of Norwalk (rNV), Mexico (rMxV), Hawaii (rHV), and Grimsby viruses (rGrV). VA98387 shared 98% aa identity with GrV; rVA98387 was detected by antisera to GrV. VA98115 shared 87% aa identity with Desert Storm virus and 65% aa identity with prototype Norwalk virus (NV); rVA98115 reacted weakly with NV antisera. VA97207 shared 80% aa identity with Amsterdam and 75% aa identity with Leeds strains and rVA97207 was not detected by any of the heterologous antibodies. In conclusion, VA97207 and VA98115 may belong to CV antigenic types not previously expressed, while VA98387 is a GrV-like virus. Low levels of cross-reactive antibodies were detected between types. Further studies to characterize these antigens and to develop enzyme immune assays (EIAs) for these strains are in progress. PMID- 11855627 TI - Phylogenetic and serological characterization of echovirus 11 and echovirus 19 strains causing uveitis. AB - The strains of echovirus 19 (EV 19) and echovirus 11 (EV11), isolated from infants with similar clinical symptoms of acute enterovirus uveitis (EU) in Russia (Siberia) in 1980-1989, were investigated phylogenetically (nucleotide sequence of a 300 nt fragment in 5' NTR and VP4 junction) and serologically. The result confirmed that viruses belong to the Enterovirus genus, with 58-80% nt sequence homology with previously sequenced enteroviruses, and showed the genetical identity between the strains isolated during each of five outbreaks of the EU. The results also demonstrated that isolates from the last three outbreaks of EU belong to the same phylogenetic group despite the remarkable spatial and temporal distance between the outbreaks. The results confirm the role of these echoviruses in the etiology of the EU. Based on phylogenetic and serological comparisons the studied strains were divided into three distinct groups: group I, EV19/K (Krasnoyarsk, 1980-1981), group II, EV11/A (Krasnoyarsk, 1982), group III, EV11/B (Krasnoyarsk, 1986; Omsk, 1987-1988; Irkutsk, 1989). Minor details of the epidemiology of the outbreaks were also revealed. PMID- 11855628 TI - Prevalence of, and antigenic variation in, serotype G10 rotaviruses and detection of serotype G3 strains in diarrheic calves: implications for the origin of G10P11 or P11 type reassortant asymptomatic strains in newborn children in India. AB - Previous studies have shown predominant association of G10P11 type bovine rotavirus-derived reassortant strains with asymptomatic infections in newborn children in India. To understand the epidemiological and genetic basis for the origin of these strains in humans, the relative frequencies of different serotypes among bovine rotaviruses (BRVs) isolated from southern, western and central regions of the country were determined by subgroup and serotype analysis as well as nucleotide (nt) sequence analysis of the genes encoding the outer capsid proteins VP4 and VP7. Since the human G10P11 asymptomatic neonatal strain I321 possessed NSP1 from a human rotavirus, to determine its genetic origin in the bovine strains, comparative analysis of partial gene sequences from representative G10P11 strains was also carried out. The following observations were of great epidemiological significance, (i) G10P11 strains predominated in all the three regions with frequencies ranging between 55.6% and 85.2%. In contrast to the high prevalence of G6 strains in other countries, only one G6 strain was detected in this study and G8 strains represented 5.8% of the isolates, (ii) among the G10 strains, in serotyping ELISA, four patterns of reactivity were observed that appeared to correlate with the differences in electropherotypic patterns and amino acid (aa) sequence of the VP7, (iii) surprisingly, strains belonging to serotype G3 were detected more frequently (10.7%) than those of serotypes G6 and G8 combined, while strains representing the new serotype (G15) were observed in a single farm in Bangalore, and (iv) about 3.9% of the isolates were nontypeable as they exhibited high cross reactivity to the serotyping MAbs used in the study. Comparative analysis of the VP7 gene sequence from the prototype G3 MAb-reactive bovine strain J63 revealed greatest sequence relatedness (87.6% nt and 96.0% aa) with that of serotype G3 rhesus-monkey strain RRV. It also exhibited high sequence homology with the VP7 from several animal and animal rotavirus-related human G3 strains (Simian SA11; equine ERV316 and FI-14; canine CU-1 and K9; porcine 4F; Feline Cat2 and human HCR3, YO and AU1). Partial nucleotide sequence analysis of the NSP1 gene of J63 showed greatest nt sequence homology (95.9%) to the NSP1 gene allele of the Indian G8 strain, isolated from a diarrheic child, which is likely to have been transmitted directly from cattle and 92.6% homology to that of the bovine G8 strain A5-10 suggesting the likely origin of J63 by gene reassortment between a bovine G8 strain and a G3 animal strain. Prevalence of G10P11 strains in cattle and G10P11 or P11 type reassortant strains in asymptomatic neonates as well as detection of G8P[1] strains in diarrheic children support our hypothesis for bidirectional transmission of rotaviruses between humans and cattle and origin of novel strains catalyzed by the age-old traditions and socio-economic conditions in India. PMID- 11855629 TI - Herpes simplex type 1 infects and establishes latency in the brain and trigeminal ganglia during primary infection of the lip in cotton rats and mice. AB - The majority of the human population has been infected with herpes simplex virus type 1 (HSV-1). During a typical primary episode, HSV-1 spreads from the oral pharynx to the trigeminal ganglia, where a latent HSV-1 infection is established. Cold sores at the mucocutaneous junction of the lip are the typical manifestation of recurrent HSV-1. We investigated whether HSV-1 also infects the brain during the primary infection. We used HSV-1 infected BALB/c mice and inbred cotton rats as models. While both species were susceptible to HSV-1 infection, the time course of lesion formation and healing in the cotton rat more closely reflected that seen in humans. In both species, HSV-1 replicated in the brainstem and cerebellum, as well as the trigeminal ganglia during a primary infection of the lip. The brain infection was produced by a low inoculation dose, and did not cause observable neurologic signs or mortality. Using PCR and RT-PCR techniques, we demonstrated HSV-1 thymidine kinase in the absence of infectivity in the brains of both species 30-40 days after primary infection. PMID- 11855630 TI - Molecular epidemiology of TTV-like mini virus in Norway. AB - TT virus (TTV), the first human circovirus to be discovered, appears to be present in most people; less is known about the prevalence of the related TTV like mini virus (TLMV). A sensitive nested PCR, specific for TLMV, detected the virus in 48% of 201 sera (Norwegian blood donors) previously found to have a 90% prevalence of TTV. More samples were either positive for both or negative for both viruses than what would have been expected from a random distribution (p = 0.08). Sequence analysis revealed considerable heterogeneity of Norwegian TLMV as compared to international sequences, suggesting that TLMV is efficiently dispersed in human populations. PMID- 11855631 TI - Critical involvement of CD40 in protection against herpes simplex virus infection in a murine model of genital herpes. AB - We studied the requirement for CD40+ cells in the resolution of vaginal infection with avirulent herpes simplex virus type I (HSV-1) in vivo using CD40-deficient mice, which were susceptible to infection with avirulent HSV-1. Compared with wild-type mice, CD40-deficient mice could not eliminate HSV-1 virus effectively from the vaginal mucosa and produced lower amounts of interleukin-12 and interferon-gamma. These results show that the induction and activation of CD40+ cells are important for HSV prevention, facilitating the activation of T cells to induce an efficient HSV clearance from the vaginal mucosa and to prevent lethal illness due to HSV infection. PMID- 11855632 TI - The measles virus hemagglutinin downregulates the cellular receptor SLAM (CD150). AB - Signaling lymphocyte activation molecule (SLAM), a cellular receptor for measles virus, was downregulated from the surface of cells infected with either the Edmonston or wild-type KA strain of measles virus. Transfection of the expression plasmid encoding the Edmonston or KA hemagglutinin, but not the fusion protein, induced downregulation of SLAM in not only cells expressing the envelope protein on the surface, but those not expressing it. After cocultivation with cells expressing the hemagglutinin, SLAM-expressing cells also exhibited downregulation of SLAM. Thus, the measles virus hemagglutinin can induce downregulation of SLAM in cells either expressing or coming in contact with it. PMID- 11855633 TI - Analysis of the entire genomes of thirteen TT virus variants classifiable into the fourth and fifth genetic groups, isolated from viremic infants. AB - TT virus (TTV) DNA in serum samples obtained from 24 TTV-infected infants was amplified by polymerase chain reaction (PCR) with inverse primers derived from the untranslated region. The amplified PCR products were molecularly cloned; six clones each were analyzed. Seventy-six (53%) of the 144 TTV clones were classified into group 4 (YONBAN isolates), and 22 (15%) into a novel genetic group (group 5). The TTV clones in group 4 were classified into 9 types, and those in group 5 into 4 types. The entire nucleotide sequence of one representative clone each from the 13 types were determined; they comprised 3570 3770 nucleotides, and had poor homology to TTVs of groups 1-3 (TA278, PMV and SANBAN isolates). A phylogenetic tree based on the entire nucleotide sequence of open reading frame 1 confirmed the presence of five distinct clusters separated by a bootstrap value of 100%. Analysis of 13 TTV variants demonstrated preservation of the genomic organization and transcription profile in all TTV groups. TTV group 4 was detected in 54% or 72% of 7-to-12-month-old infants in Japan and China, respectively, which is comparable with that among adults in the respective country, indicating early and frequent acquisition of this TTV group in infancy. PMID- 11855634 TI - Clone B7 cells have a single copy of SIVsmB7 integrated in chromosome 20. AB - B7 is the designation of a cell clone derived from the human cell line CEMx174, which was infected with SIVsmH3 clone. B7 cells chronically produce high quantities of non-infectious virus-like particles (VLP) denominated SIVsmB7. Here we report the molecular characterization of the B7 cell line. We found that B7 cells have a single copy of the SIVsmB7 provirus integrated in a noncoding region of chromosome 20 (nt 24,957 of clone RP5-963K23 on human chromosome 20q 13.11 13.2). Similarly to HIV and SIVmac, we show that integration of SIVsm results in a characteristic five base pair sequence repeat of host DNA that flanks the proviral DNA genome. Since the SIVsmB7 genome has a deletion in the IN coding sequence, the generation of this defective proviral genome most likely occurred during a faulty process of reverse transcription. Thus, these studies reveal the molecular clonality of the SIVsmB7 VLP produced by B7 cells. These genetically homogeneous VLP are useful reagents for vaccine development. In addition, these particles have been used by others (Montelaro et al.) to study the maturation of immune system responses to SIV infection. PMID- 11855635 TI - Kinetics of porcine circovirus type 2 replication. AB - The kinetics of porcine circovirus type 2 (PCV2) replication in PK15 cells was examined. During productive infection, viral antigens, RNA transcripts and progeny viruses all increased in a time dependent manner. Viral antigens were observed in a few cells at 18 hour postinfection (h p.i.) and cell-free progeny viruses began to appear at about 30 h p.i. Viral transcripts were detected by 18 h p.i. and the capsid protein RNA of 950 nucleotides (nt) was the most abundant RNA species. Two other RNAs of sizes 750 and 450 nt, derived from the predicted replication associated protein (Rep) gene region, were also detected. These two RNAs share 3' common nucleotide sequences and they are transcribed in the same orientation as the proposed unspliced Rep RNA or the recently described Rep' RNA. The 35 kD capsid protein was observed at 30 h p.i. by Western blot analysis and it appeared to be the most immunodominant protein in swine exposed to PCV2. The capsid proteins of PCV type 1 and PCV2 each contain a nuclear localization signal sequence capable of targeting a reporter protein to the nucleoli of transfected cells when the capsid proteins were expressed as 3' fusion polypeptides. Although previous reports indicated that PCV2 capsid proteins localized predominantly in the nuclei of infected cells, we observed an abundant amount of PCV2 capsid proteins in the cytoplasm of many cells of the infected cultures. The cells that exhibited cytoplasmic capsid proteins also contained virus nucleic acids, indicating that these proteins were synthesized by the infected cells and not through uptake from the culture medium. Elucidation of the changes that affect the localization pattern of PCV2 capsid proteins, nuclear versus cytoplasmic, requires further investigation. PMID- 11855636 TI - Evolutionary and taxonomic implications of conserved structural motifs between picornaviruses and insect picorna-like viruses. AB - A comparison of the recently determined structure of an insect picorna-like virus, Cricket paralysis virus (CrPV), with that of the mammalian picornaviruses shows that several structural features are highly conserved between these viruses. These conserved features include the topology of the coat proteins, the conformation of most loops, and the general arrangement of the internally located N-terminal arms of the coat proteins. The conformational conservation of the N termini of the three major coat proteins between CrPV and the picornaviruses suggests a putative ancestral T = 3 virus. Comparisons of the genome structure and amino-acid sequence of the coat proteins of CrPV with a number of other insect picorna-like viruses show that most of them belong to a novel group, recently given the interim name Cricket paralysis-like viruses. Two other insect picorna-like viruses, Infectious flacherie virus (IFV) and Sacbrood virus (SBV), for which the genome sequences have recently been determined, have very different coat protein sequences and a genome organization more like the picornaviruses. However, the position of the small VP4 protein in the structural protein polyprotein as well as the mechanism for its cleavage from VP3 upon assembly strongly suggests an evolutionary link to the "Cricket paralysis-like viruses". We propose that the picornaviruses, Cricket paralysis-like viruses and IFV/SBV group are a natural assemblage. The ancestor for this assemblage had a structure based upon the CrPV/picornavirus paradigm and a genome encoding a single major coat protein; gene duplication and rearrangements have subsequently produced the viruses that we observe today. We also discuss the possible relatives of the proposed assemblage and the likely implications of future structural studies that may be carried out on the putative relatives. PMID- 11855637 TI - Genetic diversity in the VP1 gene of foot-and-mouth disease virus serotype Asia 1. AB - Complete nucleotide sequence of the 1D (VP1-encoding) gene of 61 foot-and-mouth disease (FMD) serotype Asia I virus isolates recovered from different outbreaks in India between 1985 and 1999 including two vaccine strains currently used were determined. The sequences were compared with each other and those from other Asian countries. On the basis of phylogenetic analysis the viruses could be grouped into four genotypes (genotypes I-IV). All the 61 isolates from India belong to a single genotype (genotype-II) which is further subdivided into three lineages (B1, B2 and B3) under the same genotype. The viruses of the lineage B1 and B3 were found to be more prevalent before 1996 while the viruses of lineage B2 appeared to be new variants responsible for most of the recent outbreaks. Most of the isolates of lineage B1 lack one amino acid in the VP1 protein (position 44) whereas most of the isolates of lineage B2 and B3 contain it which indicates the possibility of these lineages having evolved independently. The rate of evolution of FMDV Asia 1 virus was also estimated and found to be 2.7 x 10(-2) synonymous substitutions per nucleotide per year. PMID- 11855638 TI - Circadian rhythms in Arabidopsis: time for nuclear proteins. AB - To be prepared for the periodic alterations of light and darkness in their environment, plants utilise an endogenous clock to anticipate these changes and to time physiological processes appropriately. Large-scale screens for mutants with perturbed circadian output rhythms, as well as characterisation of mutants with altered flowering time in Arabidopsis thaliana (L.) Heynh., have led to advances in our knowledge of the molecular basis of this internal timing system. The repertoire of proteins, mostly nuclear-localised, that play a crucial role in clock regulation, as well as of the photoreceptors involved in transmitting light information to synchronise the endogenous clock with the outside world, has expanded considerably and is the subject of this review. PMID- 11855639 TI - Multifunctionality of plant ABC transporters--more than just detoxifiers. AB - The ABC-transporter superfamily is one of the largest protein families, and members can be found in bacteria, fungi, plants and animals. The first reports on plant ABC transporters showed that they are implicated in detoxification processes. The recent completion of the genomic sequencing of Arabidopsis thaliana (L.) Heynh. [Arabidopsis Genome Initiative (2000) Nature 408:796-815] showed that Arabidopsis contains more than 100 ABC-type proteins; 53 genes code for so-called full-size transporters, which are large proteins of about 150 kDa consisting of two hydrophobic and two hydrophilic domains. The large number of genes in the MDR/MRP and PDR5-like sub-clusters and the strong sequence homology found in many cases suggest functional redundancy. One reason for the high number of genes can be attributed to the duplication of large segments of Arabidopsis chromosomes. Recent results indicate that the function of this protein family is not restricted to detoxification processes. Plant ABC transporters have been demonstrated to participate in chlorophyll biosynthesis, formation of Fe/S clusters, stomatal movement, and probably ion fluxes; hence they may play a central role in plant growth and developmental processes. PMID- 11855640 TI - High-level production of the non-cariogenic sucrose isomer palatinose in transgenic tobacco plants strongly impairs development. AB - Palatinose (isomaltulose, 6-O-alpha-D-glucopyranosyl-D-fructose) is a structural isomer of sucrose which is produced from sucrose by some bacterial strains as a reserve material during periods of low carbon availability. The ability to synthesise palatinose is not only advantageous for the bacteria but is also of industrial interest since palatinose is used as a sucrose substitute in food production. To explore the possibility of palatinose production in plants a recently isolated sucrose isomerase gene (palI; EC 5.4.99.11) from Erwinia rhapontici [F. Bornke et al. (2001) J Bacteriol 183: 2425-2430] was cloned into a plant expression vector between the constitutive 35S CaMV promoter and the octopine synthase polyadenylation signal. To allow secretion of the protein into the apoplast the signal peptide of the potato proteinase inhibitor II was N terminally fused to the pall coding region. Expression of the protein was verified by northern and western blot analyses. Efficient secretion of the protein was demonstrated by palI detection in intercellular fluids. Transgenic plants expressing palI accumulated high levels of palatinose. As a consequence, transgenic plants showed severe phenotypic alterations. Young leaves were curled and developed bleached areas during maturation. Flowers were misshapen and sterile. Based on nonaqueous fractionation experiments palatinose was found in several subcellular compartments, indicating limited membrane transport of the sugar. In contrast to results obtained with short-term feeding experiments, no evidence for palatinose-mediated regulation of photosynthetic or defence genes could be obtained in the transgenic palI-expressing tobacco plants. Based on our results we conclude that plants can efficiently be used as bioreactors for the production of palatinose. Furthermore, tissue-specific expression of palI should allow carbon allocation to specific tissues and/or cell-types to be modulated. PMID- 11855641 TI - MADS-box gene expression in lateral primordia, meristems and differentiated tissues of Arabidopsis thaliana roots. AB - Although MADS-box genes involved in flower and fruit development have been well characterized, the function of MADS-box genes expressed in vegetative structures has yet to be explored. At least seven members of this family are grouped in clades of genes that are preferentially expressed in roots of Arabidopsis thaliana (L.) Heynh.. We report here the cloning of the AGL21 MADS-box gene, which belongs to the ANR1 clade, and the mRNA in situ expression patterns of this and two other root MADS-box genes. AGL17 appears to be a lateral root cap marker in the root tip, and towards the elongation zone this gene is expressed in the epidermal cells. AGL21 is highly expressed in lateral root primordia and it has a punctate expression pattern in the primary root meristem. AGL12 also has a punctate expression pattern in the primary root meristem. AGL12 and AGL21 are also expressed in the central cylinder of differentiated roots and both are expressed in developing embryos. This study, combined with previous phylogenetic analyses, indicates that these MADS-box genes may play distinct regulatory roles during root development. PMID- 11855642 TI - The Arabidopsis gene PROLIFERA is required for proper cytokinesis during seed development. AB - The Arabidopsis thaliana (L.) Heynh. gene PROLIFERA (PRL) is a member of the MCM family of genes that are required for DNA replication during the S phase of the cell cycle. PRL is expressed in dividing cells throughout plant development. During reproductive development, PRL is expressed in both the developing megaspore mother cells and microspore mother cells, but is not expressed in the developing microgametophyte, suggesting that it does not function in the final haploid divisions leading to the production of a mature pollen grain. Disruption of PRL leads to megagametophyte and embryo lethality. prl mutant embryos arrest at a variety of stages, and often show defects in cytokinesis. Multinucleate cells and non-stereotypical cell division planes are commonly observed in developing prl mutant embryos, although mcm mutations in other organisms have not been reported to affect cytokinesis. These observations suggest that PRL may play a role in cytokinesis that is distinct from its role in regulating DNA replication. Additionally, a novel cytokinesis checkpoint that monitors cell cycle progression may exist in Arabidopsis. PMID- 11855643 TI - Hydroponically cultivated radish fed L-galactono-1,4-lactone exhibit increased tolerance to ozone. AB - Leaf L-ascorbate content of an ozone (O3)-sensitive radish genotype (Raphanus sativus L. cv. Cherry Belle) was increased 2-fold by feeding hydroponically cultivated plants L-galactono- 1,4-lactone (GalL). Plants were grown in controlled-environment chambers ventilated with charcoal/Purafil-filtered air, and administered one of two O3 fumigation regimes: chronic exposure (75 nmol O3 mol(-1) for 7 h day(-1) for 21 days) and acute exposure (180 nmol O3 mol(-1) for 9 h). Chronic O3 exposure decreased root growth by 11% in plants maintained in pure nutrient solution (-GalL), but resulted in no change in root growth in GalL fed plants (+GalL). Similarly, GalL-feeding counteracted the negative effects of O3 on CO2 assimilation rate observed in control plants (-GalL). Under acute O3 exposure, GalL-fed plants showed none of the visible symptoms of injury, which were extensive in plants not fed GalL. Leaf CO2 assimilation rate was decreased by acute 03 exposure in both GalL treatments, but the extent of the decline was less marked in GalL-fed plants. No significant changes in stomatal conductance resulted from GalL treatment, so O3 Uptake into leaves was equivalent in + GalL and -GalL plants. Feeding GalL, on the other hand, enhanced the level of ascorbate, and resulted in the maintenance of the redox state of ascorbate under acute O3 fumigation, in both the leaf apoplast and symplast. The effect of GalL treatment on ascorbate pools was consistent with the reduction in O3 damage observed in GalL-fed plants. Attempts to model O3 interception by the ascorbate pool in the leaf apoplast suggested a greater capacity for O3 detoxification in GalL-fed plants, which corresponded with the increase in O3 tolerance observed. However, modelled data for GalL-fed plants suggested that additional constituents of the leaf apoplast may play an important role in the attenuation of environmentally-relevant O3 fluxes. PMID- 11855644 TI - Water deficit during root development: effects on the growth of roots and osmotic water permeability of isolated root protoplasts. AB - The effect of low water potentials on root growth of flax (Linum usitatissimum L. cv. Ariane), rape (Brassica napus L. de Candolle, cv. Bristol), hard wheat (Triticum turgidum L. cv. Cham1) and soft wheat (Triticum aestivum L. cv. Ritmo) was studied by measuring the osmotic water permeability (Pos) of root protoplasts and the protein abundance of PIP1 and PIP2 aquaporins. These different species require more or less water, the most sensitive to water deficit being flax and rape. Ritmo, is a cultivar of wheat adapted to temperate zones, while the other cultivar Cham1 is adapted to low-rainfall areas. The seedlings were germinated and grown in water, salt or sugar solutions at different water potentials. The values of Pos for flax, rape and Chaml wheat were normally distributed and could be characterized by mean +/- SD. Root protoplasts from water-grown seedlings had Pos values of 485+/-159 microm s(-1) (flax), 582+/-100 microm s(-1) (rape), and 6.3+/-3.5 microm s(-1) (Cham1). At the same age, the protoplasts from Ritmo exhibited a much wider range of values than the protoplasts of Cham1. When seedlings were grown under conditions of osmotic or salt stress, the mass of the roots was reduced for all species. With 0.25 mol kg(-1) sorbitol or 0.125 M NaCl, the Pos for flax, rape and Cham1 remained constant or slightly increased, while for Ritmo the reduction in the mass of the roots was paralleled by a reduction in Pos. Only Cham1 and Ritmo were able to germinate at a lower potential (0.5 mol kg(-1) sorbitol). For Ritmo the reduction in the mass of the roots was paralleled by a reduction in Pos when grown in this stress condition and both wheats exhibited low Pos values. The expression of the PIP1 and PIP2 aquaporins families was also studied by immunoblotting. We did not observe any difference in protein expression for any of the species, whatever the growing conditions. We suggest that the high Pos values for flax and rape could play a role in the sensitivity of these plants at low water potential. The low native Pos for Cham1 in spite of the expression of both families of aquaporins may reflect its adaptation to low rainfall conditions by a functional regulation of the water channels. For a similar reason, the low-water-potential-induced Pos of Ritmo may also correspond to a down-regulation of the aquaporins, reflecting adaptation of this wheat to water-deficit conditions. PMID- 11855645 TI - A 45-kDa protein kinase related to mitogen-activated protein kinase is activated in tobacco cells treated with a phorbol ester. AB - Phorbol 12-myristate 13-acetate (PMA), a potent activator of protein kinases in animals, elicits the transient activation of a 45-kDa protein kinase in tobacco cell-suspension cultures. The 45-kDa protein kinase preferentially phosphorylates myelin basic protein (MBP), a general substrate for MAPK. Studies using cycloheximide indicated that protein synthesis is not required for the activation of the kinase. Treatment of tobacco cell extracts containing the activated kinase with either serine/threonine-specific or tyrosine-specific protein phosphatase abolished the kinase activity, which consequently appears to be regulated by phosphorylation. By using an immune complex kinase assay with antibodies specific for stress-responsive MAPKs, we show that the PMA-activated kinase is immunologically related to the wound-induced protein kinase (WIPK), and not to the salicylic acid-induced protein kinase (SIPK), two representative members of the tobacco MAPK family, known to be activated by extracellular stimuli. Furthermore, the activated kinase was recognized by phospho-specific MAPK antibodies. Collectively, these results indicate that phorbol ester promotes the activation of a 45-kDa protein kinase related to WIPK in tobacco cells. Activation of WIPK in response to PMA is associated with protein phosphorylation but not with an increase in protein level. PMID- 11855646 TI - Evidence that gamma-aminobutyric acid is a major nitrogen source during Cladosporium fulvum infection of tomato. AB - The growth of the biotrophic pathogen Cladosporium fulvum within the tomato (Lycopersicon esculentum Mill.) leaf is restricted to the intercellular space. Previous studies from this laboratory have demonstrated that gamma-aminobutyric acid (GABA) accumulates to millimolar concentrations in the apoplast during a compatible interaction. We decided to further investigate the role of GABA during infection. A gene encoding a required enzyme for GABA metabolism, GABA transaminase (Gat1), was cloned and sequenced from C. fulvum. The predicted protein sequence of Gat1 had high homology to other fungal GABA transaminases, particularly from Aspergillus nidulans. In vitro expression experiments revealed Gat1 to be strongly expressed during fungal growth on both GABA and glutamate whereas nearly no expression was evident during nitrogen starvation conditions. Expression of Gat1 was also apparent during infection, suggesting for the first time that C. fulvum actively metabolises GABA during infection. This indicates that the fungus may be utilising the GABA in the apoplast as a nutrient source. Further analysis revealed that the expression of tomato glutamate decarboxylase, the enzyme responsible for GABA synthesis, appeared appreciably higher during a compatible interaction than in the incompatible interaction. These findings imply that the infecting fungus may alter the physiology of the tomato leaf with the result that a source of nitrogen is supplied. PMID- 11855647 TI - An isoleucine-leucine substitution in chloroplastic acetyl-CoA carboxylase from green foxtail (Setaria viridis L. Beauv.) is responsible for resistance to the cyclohexanedione herbicide sethoxydim. AB - The cDNAs encoding chloroplastic acetyl-CoA carboxylase (ACCase, EC 6.4.1.2) from three lines of Setaria viridis (L. Beauv.) resistant or sensitive to sethoxydim, and from one sethoxydim-sensitive line of Setaria italica (L. Beauv.) were cloned and sequenced. Sequence comparison revealed that a single isoleucine-leucine substitution discriminated ACCases from sensitive and resistant lines. Using near isogenic lines of S. italica derived from interspecific hybridisation, we demonstrated that the transfer of the S. viridis mutant ACCase allele into a sethoxydim-sensitive S. italica line conferred resistance to this herbicide. We confirmed this result using allele-specific polymerase chain reaction and showed that a single copy of the mutant allele is sufficient to confer resistance to sethoxydim. We conclude that a mutant allele of chloroplastic ACCase encoding a leucine residue instead of an isoleucine residue at position 1780 is a major gene of resistance to sethoxydim. PMID- 11855648 TI - ADP-glucose pyrophosphorylase from wheat endosperm. Purification and characterization of an enzyme with novel regulatory properties. AB - ADP-glucose pyrophosphorylase (AGPase; EC 2.7.7.27) was purified and characterized from two wheat (Triticum aestivum L.) tissues: leaf and endosperm. The leaf enzyme, purified over 1,300-fold, was found to be a heterotetramer composed of subunits of 51 and 54 kDa and possessing regulatory properties typical of AGPases from photosynthetic tissues, being mainly regulated by 3 phosphoglycerate (activator; A0.5=0.01 mM) and orthophosphate (inhibitor; I0.5=0.2 mM). Conversely, the enzyme from wheat endosperm was insensitive to activation by 3-phosphoglycerate and other metabolites. It was, however, inhibited by orthophosphate (I0.5=0.7 mM), ADP (I0.5=3.2 mM) and fructose-1,6 bisphosphate (0.5 = 1.5 mM). All of these inhibitory actions were reversed by 3 phosphoglycerate and fructose-6-phosphate. The endosperm enzyme was found to be a heterotetramer composed of subunits of 52 and 53 kDa, which were recognized by antiserum raised to spinach leaf AGPase. The results suggest that wheat endosperm AGPase possesses distinctive regulatory properties that are relevant in vivo. PMID- 11855649 TI - Characterization of three new members of the Arabidopsis thaliana calmodulin gene family: conserved and highly diverged members of the gene family functionally complement a yeast calmodulin null. AB - Three genes encoding members of the EF-hand family of Ca2+-binding proteins were identified from Arabidopsis thaliana (L.) Heynh. sequences deposited in the expressed sequence tag and genomic sequence databases. Full-length cDNAs for each of the genes, Cam7, Cam8, and Cam9, were sequenced. Cam7 encodes a conventional 16.8-kDa, 148-amino-acid calmodulin protein (CaM). In contrast, Cam8 and 9 encode highly diverged isoforms of the protein that share 73 and 49% amino acid sequence identity, respectively, with CaM7. RNA gel blot and reverse transcription polymerase chain reaction experiments revealed that each of the genes is expressed in leaves, flowers and siliques. To test the functional properties of the polypeptides encoded by these genes, they were expressed in Escherichia coli and the yeast Saccharomyces cerevisiae. Each was purified by Ca2+-dependent hydrophobic affinity chromatography. CaM7, but neither CaM8 nor CaM9, formed a complex with a basic amphiphilic helical peptide in the presence of Ca2+ that could be identified by gel electrophoresis. In spite of these in vitro differences, each of the sequences functionally substituted for yeast CMD1 to maintain viability. Isolation of yeast strains complemented by Cam9 required selection against the plasmid harboring wild-type yeast sequences, whereas complementation by Cam7 and Cam8 did not. These results suggest that the mechanism of action of CaM8 and CaM9 is similar to that of more conventional CaM sequences. CaM9, and to a lesser degree CaM8, however, appear to represent Ca2+ binding sensor proteins that interact with a more limited set of target proteins than do more conventional CaM isoforms. PMID- 11855650 TI - The effect of physical and chemical treatment on the mechanical properties of the cell wall of the alga Chara corallina. AB - Single large internode cells of the charophyte (giant alga) Chara corallina were dissected to give sheets of cell wall, which were then notched and their mechanical properties in tension determined. The cells were subjected to a thermal treatment in excess water (cf. cooking), which had little effect on strength but increased the stiffness, contrasting with the behaviour of higher plant tissues. Extraction in CDTA (cyclohexane-trans-1,2-diamine-N,N,N',N' tetraacetate) or 4 M KOH reduced the strength from 17 MPa to 10 MPa, although sequential extraction in CDTA and 4 M KOH reduced the strength further to 4 MPa. The stiffness decreased from 500 MPa to 300 MPa on extraction in CDTA or 4 M KOH, while falling to 70 MPa after extraction in CDTA followed by 4 M KOH. Conventional sequential extraction in CDTA, Na2CO3 at 1 degrees C and 20 degrees C, and KOH at 0.5 M, 1 M, 2 M and 4 M caused a gradual decrease in stiffness and strength after the CDTA treatment to the same lower values. This result is in keeping with mechanical properties for plant tissues, but in contrast to the removal of pectic polysaccharides from model cell wall systems, which does not reduce the stiffness. PMID- 11855651 TI - Overexpression of violaxanthin de-epoxidase: properties of C-terminal deletions on activity and pH-dependent lipid binding. AB - Violaxanthin de-epoxidase (VDE) is localized in the thylakoid lumen and catalyzes the de-epoxidation of violaxanthin to form antheraxanthin and zeaxanthin. VDE is predicted to be a lipocalin protein with a central barrel structure flanked by a cysteine-rich N-terminal domain and a glutamate-rich C-terminal domain. A full length Arabidopsis thaliana (L.) Heynh. VDE and deletion mutants of the N- and C terminal regions were expressed in Escherichia coli and tobacco (Nicotiana tabacum L. cv. Xanthi) plants. High expression of VDE in E. coli was achieved after adding the argU gene that encodes the E. coli arginine AGA tRNA. However, the specific activity of VDE expressed in E. coli was low, possibly due to incorrect folding. Removal of just 4 amino acids from the N-terminal region abolished all VDE activity whereas 71 C-terminal amino acids could be removed without affecting activity. The difficulties with expression in E. coli were overcome by expressing the Arabidopsis VDE in tobacco. The transformed tobacco exhibited a 13- to 19-fold increase in VDE specific activity, indicating correct protein folding. These plants also demonstrated an increase in the initial rate of nonphotochemical quenching consistent with an increased initial rate of de epoxidation. Deletion mutations of the C-terminal region suggest that this region is important for binding of VDE to the thylakoid membrane. Accordingly, in vitro lipid-micelle binding experiments identified a region of 12 amino acids that is potentially part of a membrane-binding domain. The transformed tobacco plants are the first reported example of plants with an increased level of VDE activity. PMID- 11855652 TI - Plasmalemma protection by the apoplast as assessed from above-zero ozone concentrations in leaf intercellular air spaces. AB - To evaluate reactive absorption of ozone (O3) in the leaf apoplast, amphistomatous leaves of Phaseolus vulgaris L. were allowed to take up O3 through the stomata on the lower leaf surface at high rates for 3-5 min. Up to 5% of the O3 taken up diffused through the leaf and emerged from the stomata on the upper surface, suggesting above-zero O3 concentrations in the leaf intercellular air space, [O3]i. Moreover, measurements revealed that [O3]i increased during exposure to the pollutant. Time patterns of O3 fluxes through the gas phase and into the aqueous apoplast indicated that the increase in [O3]i was the result of a decrease in the diffusion-reaction conductance of the aqueous apoplast, gaq. Under an intense O3 pulse, gaq approached the value of the pure diffusional conductance within 2.5 min of exposure, suggesting the exhaustion of protective resources in the leaf apoplast. Toward the end of the exposure gaq tended to increase, suggesting either a recovery in the protective resources in the leaf apoplast and/or the induction of new defences. The possibility of estimating the degree of protection afforded by apoplast constituents and the rate of recovery of these protective systems in intact leaves using brief O3 pulses is discussed. PMID- 11855653 TI - The stability of the Chlorella nitrate reductase mRNA is determined by the secondary structure of the 5'-UTR: implications for posttranscriptional regulation of nitrate reductase. AB - Nitrate reductase (NR; EC 1.6.1.1-3) can be controlled at both transcriptional and posttranscriptional levels. Here we describe stability of NR mRNA as a mechanism of control. The NR gene in Chlorella vulgaris (Warburg strain) transcribes a stable mRNA and an unstable mRNA. In-vitro-synthesized transcripts representing these mRNAs show the same stability characteristics. The unstable mRNA is 30 nucleotides longer at the 5'-UTR compared to the stable mRNA. Using an RNA-folding program the 5'-UTR of the longer unstable RNA showed a more extensive stem-loop structure compared to the more linear form of the shorter stable mRNA. Transcripts representing RNAs with intermediate 5'-UTRs folded similarly to the long form and were unstable, or similarly to the short form and were more stable. Thus the secondary structure of the 5'-UTR of NR mRNA is important in the stability of NR transcripts in Chlorella and allows the cell to respond to changes in nitrogen source in an energy-efficient manner. PMID- 11855654 TI - Substrate specificity and sequence analysis define a polyphyletic origin of betanidin 5- and 6-O-glucosyltransferase from Dorotheanthus bellidiformis. AB - Betanidin 6-O-glucosyltransferase (6-GT) is involved in the glycosylation of betacyanins, which replace the chromogenic anthocyanins as flower colorants in the Caryophyllales. The 6-GT cDNA was cloned from a cDNA library of Dorotheanthus bellidiformis (Burm. f.) N.E. Br., and the amino acid and nucleotide sequences were shown to be distinctly different from the corresponding betanidin 5-O glucosyltransferase (5-GT) from the same plant species. Although both enzymes share very similar substrates, the proteins show only 19% amino acid sequence identity. In contrast, the protein sequence of the 6-GT showed significant identity to GTs from other species and may identify a new cluster of putative anthocyanidin GTs. Therefore, 6-GT and 5-GT apparently have evolved independently from ancestral glucosyltransferases involved in flavonoid biosynthesis. PMID- 11855655 TI - Positive inotropes--a new horizon or still a dead end? PMID- 11855656 TI - Na+ channel activators as positive inotropic agents for the treatment of chronic heart failure. AB - The Na+ channel agonists DPI 201-106, BDF 9148 and BDF 9198 are a new group of positive inotropic agents which increase cardiac contractility in a cAMP independent manner. The most likely mechanism by which positive inotropy is mediated is an enhancement of Na+/Ca2+ exchange activity in response to a Na+ channel agonist induced increase in the cardiac myocyte intracellular Na+ concentration. While the positive inotropic effect of drugs which exert their effects in a cAMP dependent manner is blunted in failing compared to nonfailing myocardium, the efficacy and potency of Na+ channel agonists is not only maintained, but enhanced in failing myocardium. This finding makes these substances interesting for the treatment of patients with heart failure. The positive inotropic effects of the Na+ channel agonists, however, are accompanied by a potential increase in the incidence of cardiac arrhythmias. These side effects might limit the clinical use of Na+ channel agonists and demand future development of Na+ channel modulators without significant arrhythmogenic effects. PMID- 11855657 TI - Phospholamban: a promising therapeutic target in heart failure? AB - Dilated cardiomyopathy and end-stage heart failure result in characteristic functional, biochemical and molecular alterations. Multiple defects in cardiac excitation-contraction coupling have been suggested to underlie disturbed myocardial function and progressive remodeling. Ca2+ uptake and release by the sarcoplasmic reticulum (SR) have been shown to be altered in various animal models and human conditions. This review will focus on SR Ca2+ ATPase and its regulatory protein, phospholamban, as potential therapeutic targets. We summarize structural and genetic approaches, which have helped to elucidate the physiological role of phospholamban as a principal regulator of cardiac contractility and beta-adrenergic stimulation in the heart. These findings are extended to the clinical arena, indicating a phospholamban/SR Ca2+ ATPase mismatch in human heart failure. Evidence is then provided, using genetically engineered mouse models, that SR dysfunction may play a key role in the onset and progression of heart failure. Phospholamban deficiency may prevent such left ventricular dysfunction and its progression to heart failure in some of the animal models with dilated cardiomyopathy. Based on these findings, we discuss the question of whether and how interfering with the phospholamban/SR Ca2+ ATPase interaction may be a promising therapeutic approach for heart failure. PMID- 11855658 TI - Mechanism of action of Ca2+ sensitizers--update 2001. AB - Ca2+ sensitizers act on the central mechanism (Ca2+ binding affinity of troponin C) and/or downstream mechanisms (thin filament regulation of actin and direct action on crossbridge cycling) of cardiac E-C coupling. Ca2+ sensitizers have mechanistic and energetic advantages over the agents that act through the upstream mechanism (intracellular Ca2+ mobilization). Ca2+ sensitizers and the agents that act through cyclic AMP-mediated signaling process have been postulated to belong to different classes, however, recent experimental findings revealed that certain Ca2+ sensitizers, such as levosimendan, OR 1896 and UD-CG 212 Cl, require cyclic AMP-mediated signaling for induction of the Ca2+ sensitizing effect. No clinically available agents act primarily via Ca2+ sensitization, but the positive inotropic effect of pimobendan and levosimendan is partly due to an increase in myofilament Ca2+ sensitivity. These agents are the hybrid of Ca2+ sensitizer and PDE III inhibitor. The extent of contribution of Ca2+ sensitizing effect of these agents to the clinical effectiveness to improve the hemodynamics in patients with heart failure is uncertain. Nevertheless pieces of evidence have been accumulating that these agents with Ca2+ sensitizing effect are clinically more effective than the agents that act purely via the upstream mechanism. PMID- 11855659 TI - Energetic stimulation of the heart. AB - The management of patients with acute heart failure includes frequently the use of positive inotropic agents in order to increase cardiac output and hence to maintain adequate tissue perfusion. The concept of energetic stimulation of the heart was invented in order to avoid deleterious effects of conventional inotropes, i.e. catecholamines and phosphodiesterase-inhibitors, on myocardial oxygen consumption and energy reserves and to circumvent potential dangerous side effects of the drugs. Metabolic support with glucose-insulin-potassium has proven efficious of improving cardiac performance and prognosis in the setting of the ischemic and postischemic heart. The glycolytic intermediate pyruvate increases contractile performance in isolated animal and human myocardium and improves short-time hemodynamics in patients with congestive heart failure, which is discussed in detail in the present article. Energetic stimulation with pyruvate could therefore be a new promising approach to the treatment of acute heart failure, if conventional therapy fails. PMID- 11855660 TI - Pharmacotherapy of dyslipidemia. AB - Reducing elevated levels of low-density-lipoprotein cholesterol (LDL-C) significantly reduces the incidence of coronary heart disease (CHD) events and mortality in hypercholesterolemic patients. CHD risk reduction is proportional to LDL-C reduction. Despite this knowledge, many physicians are not applying existing treatment guidelines to the extent required to achieve target LDL-C levels. Target LDL-C levels are not achievable for most patients without drug therapy. Based on their lipid-lowering abilities, safety, and tolerability profiles, the HMG-CoA reductase inhibitors (statins) are the first-line pharmacotherapeutic agents for hypercholesterolemia. The ability of statins to reduce CHD events and total mortality in primary- and secondary-prevention patients also supports this assertion. For combined dyslipidemia, statin monotherapy is a reasonable initial approach in patients with moderate hypertriglyceridemia because statins effectively lower both LDL-C and triglycerides. Fibrates or niacin are effective therapies for severe hypertriglyceridemia. Resins are moderately effective in isolated hypercholesterolemia, and are a useful alternative to statins in pregnant women or patients with liver disease. For severe hyperlipidemia that does not respond to single drug therapy, combination drug therapy may be required. This article reviews the various manifestations of dyslipidemia and assesses the most efficacious treatments. PMID- 11855661 TI - Antiplatelet therapy in acute coronary syndromes without persistent ST-segment elevation. AB - The treatment of ACS without persistent ST-segment elevation is evolving. Antiplatelet and antithrombin therapy forms the mainstay of medical management. The antiplatelet agents studied can be pharmacologically classified as GP lIb/IIIa receptor antagonists, ADP receptor antagonists, thromboxane inhibitors, and cyclo-oxygenase inhibitors. While aspirin, a cyclo-oxygenase inhibitor, is well entrenched in the treatment (and thus will not be reviewed here), other drugs have been subjects of intense study and large-scale clinical trials in the last decade. In this article we will explore the rationale of using antiplatelet agents, describe the platelet biology and mechanism of action of these drugs, narrate the major phase III trials, and attempt to draw conclusions from the clinical experience. Important trials which have been presented at principal international scientific meetings and which have not yet been published are also cited. PMID- 11855662 TI - Quantitation of cell migration in a rat carotid artery balloon injury model. Indications for a perivascular origin of the neointimal cells. AB - Medial and neointimal smooth muscle cells differ in phenotype. Adventitial cells have been shown to migrate to the intima and this could partly explain this difference. OBJECTIVE: The aim of this study was to quantitate the kinetics of cell migration from the adventitial and medial layers to the intima after endothelial injury. METHODS: Labelled proliferating cells at different periods post-injury in a rat carotid artery balloon injury model, were used to calculate the kinetics of migration using the alteration in cell populations and the ratio of proliferating cells. RESULTS: The increase in the number of neointimal cells was greater than the level of proliferation during the 30-day follow-up. Changes in the number and percentage of proliferating cells remained low after the appearance of the first neointimal cells. 28% of the neointimal cells were labelled during the first wave of migration, and in reverse at least 72% had migrated there. Of these migrating cells, 74% were non-proliferating. The formation of neointima was efficiently blocked with cyclophosphamide and batimastat (metalloproteinase inhibitor), which resulted in a decrease in the number of medial and intimal cells. CONCLUSION: The increase in the number of neointimal cells is mostly due to cells migrating from the outer layers through the media. The majority of these cells are not proliferating, but migration can still be efficiently blocked with antiproliferative drugs. PMID- 11855663 TI - Insulin in combination with vanadate stimulates glucose transport in isolated cardiomyocytes from obese Zucker rats. AB - Insulin stimulates glucose uptake in muscle cells via activation of protein kinase B (PKB). The protein tyrosine phosphatase (PTP) inhibitor vanadate, is a known insulin mimetic agent but the mechanism whereby vanadate exerts its effect is not clearly understood. Vanadate also has beneficial effects in the diabetic myocardium. The aim of this study was to correlate insulin stimulation of glucose uptake and PKB activation with that induced by vanadate in adult ventricular myocytes from lean and obese Zucker fa/fa rats. In lean Zucker rats, 100 nM insulin and 5 mM vanadate stimulated myocardial 2-deoxy-D-[3]glucose (2-DG) uptake from 27.17 +/- 1.72 to 96.52 +/- 10.87 and 43.86 +/- 4.02 pmole/mg protein p/30 min respectively while a combination of insulin and vanadate could not improve the maximal response of insulin. In obese Zucker hearts, basal as well as insulin and vanadate stimulated glucose uptake were severely impaired (15.49 +/- 1.44 vs 25.51 +/- 3.11 and 20.11 +/- 1.68 pmole/mg protein/30 min respectively). A combination of insulin and vanadate, resulted in a response significantly improved from the maximal response of insulin. This stimulation of 2-DG uptake was, in all instances, blocked by the PI 3-kinase inhibitors wortmannin and LY 294002. Insulin could not activate PKB, as measured by the Ser473 phosphorylated form of the enzyme, in the obese Zucker rats to the same extent as in lean controls. Similar to glucose uptake, activation of PKB by vanadate plus insulin was significantly more than that accomplished by insulin alone in obese rats. Both insulin and vanadate activation of PKB was prevented by wortmannin and LY 294002. Thus, the present study demonstrates that: (i) in cardiomyocytes from lean and obese Zucker rats, both insulin and vanadate stimulate glucose uptake and PKB activation through a PI 3-kinase sensitive pathway. (ii) In obese Zucker rats, neither insulin nor vanadate could induce glucose uptake or activation of PKB to the same extent as in lean controls. (iii) A combination of insulin with vanadate may be beneficial to increase glucose uptake in diabetic hearts, as this gives a better response than insulin alone. PMID- 11855664 TI - Short term effect of atorvastatin and vitamin E on serum levels of C3, a sensitive marker of the risk of myocardial infarction in men. AB - C3 complement is produced in response to macrophage activation and is a reliable marker of the risk of myocardial infarction in men. This study was designed to ascertain whether the treatment with atorvastatin, a powerful cholesterol lowering drug, and/or vitamin E, a natural antioxidant, may induce a short term decrease in serum C3 in subjects with persistently elevated levels. From an initial random sample of 1100 men aged 55-64 years, 140 subjects with 3 consecutive C3 measurements in the high tertile (>1.19 g/l) were selected. Those with total cholesterol <5.56 mmol/l were double blindly randomized in groups 1 (placebo, N = 28, G1) and 2 (vitamin E 600 IU/day, N= 30, G2). The subjects with total cholesterol values >5.56 mmol/l were randomized in groups 3 (placebo, N= 30, G3), 4 (atorvastatin 10 mg/day, N = 27, G4) and 5 (atorvastatin 10 mg/day + vitamin E 600 IU/day, N = 25, G5). After 3 months C3 levels were substantially unchanged in the first 4 groups, while in G5 a very significant decrement occurred: -0.070 g/l (5.2%); 95% CI 0.043-0.098; p <0.0001. "Normal" levels of C3 (< 1.19 g/l) were reached by 28% of G5 subjects. In G2 and G5 vitamin E levels increased by 60 and 36%, while in G4 they decreased by 23% (p < 0.0001), paralleling cholesterol and triglyceride fall. In all groups a progressive decrease in HDL cholesterol occurred (-17%, p < 0.0001). In conclusion, treatment with atorvastatin plus vitamin E for three months can lower persistently elevated C3 levels. PMID- 11855665 TI - Effect of cibenzoline, a class ia antiarrhythmic agent, on left ventricular diastolic function in hypertrophic cardiomyopathy. AB - We aimed to investigate whether the improvement of left ventricular (LV) diastolic function by cibenzoline, a class Ia antiarrhythmic drug, in hypertrophic obstructive cardiomyopathy (HOCM) is due to LV afterload reduction or a primary lusitropic effect on LV. Twenty-three patients with hypertrophic cardiomyopathy (11; HOCM, 12; non-obstructive HCM; HNCM) were examined. Pulsed wave Doppler, color M-mode and tissue Doppler echocardiography were performed before and 90 minutes after oral administration of cibenzoline (300 mg), and were compared with a treatment of bisoprolol (5 mg/day, 10 days). Early (E) and late diastolic LV inflow velocity, E flow propagation velocity (FPV) and early diastolic mitral annulus velocity (Ea) were measured. E/FPV and E/Ea were calculated as indices of LV filling pressure. LV outflow pressure gradients estimated using continuous-wave Doppler in HOCM markedly decreased after cibenzoline (83 +/- 42 to 40 +/- 33 mmHg, p < 0.005) and bisoprolol (44 +/- 40 mmHg, p < 0.005). Following cibenzoline, E/FPV and E/Ea were significantly decreased in both HOCM (1.75 +/- 0.53 to 1.32 +/- 0.28, p < 0.05, 18.9 +/- 6.2 to 14.8 +/- 5.0, p < 0.05, respectively) and HNCM (1.75 +/- 0.58 to 1.41 +/- 0.73, p< 0.05, 13.0 +/- 4.3 to 9.7 +/- 3.6, p< 0.01, respectively). Those in HNCM did not change by bisoprolol. Cibenzoline improved LV diastolic function in HCM, whereas bisoprolol did not affect it. Thus, the primary lusitropic effect of cibenzoline rather than LV after load reduction might have contributed to the improvement of diastolic function in HOCM. PMID- 11855666 TI - Effect of nifedipine and verapamil on the peripheral resistance of isolated perfused guinea pig hind limbs. PMID- 11855667 TI - Nitric oxide and depolarization induce hydroxyl radical generation. AB - Nitric oxide (NO) contributes to the extracellular potassium-ion concentration ([K+]o)-induced hydroxyl radical (*OH) generation. Cytotoxic free radicals such as peroxinitrite (ONOO-) and *OH may also be implicated in NO-mediated cell injury. NO is synthesized from L-arginine by NO synthase (NOS). NOS activation was induced by K+ depolarization. Oxidative modification of low-density lipoprotein (LDL) is thought to contribute to the production of oxygen derived free radicals. However, LDL oxidation may be related to noradrenaline-induced *OH generation, but LDL oxidation may be unrelated to *OH generation via NOS activation. Abnormal levels of extracellular free dopamine (DA) and/or intraneuronal Ca2+ triggered by 1-methyl-4-phenylpyridinium ion (MPP+) may be detrimental to the functioning of dopaminergic nerve terminals in the striatum. Although [K+]o-induced depolarization enhances the formation of *OH product due to MPP+, the *OH generation via NOS activation may be unrelated to the DA-induced *OH generation. Depolarization enhances the formation of *OH products via NOS activation. PMID- 11855668 TI - Long-term treatment with perindopril ameliorates dobutamine-induced myocardial ischemia in patients with coronary artery disease. AB - The present study was designed to examine whether long-term blockade of angiotensin-converting enzyme (ACE) with perindopril ameliorates dobutamine induced myocardial ischemia in patients with coronary artery disease (CAD). Twelve patients with proven CAD were randomly divided in two groups; one group received perindopril (8 mg/day, p.o.) for 3 months and another group served as a control. To evaluate anti-ischemic effects of perindopril, dobutamine stress echocardiography was performed before and 3 months after the treatment in a double-blind manner. Long-term treatment with perindopril significantly ameliorated the dobutamine-induced myocardial ischemia, as evaluated by time to the onset of symptoms, magnitude of electrocardiographic ST-segment changes, and left ventricular wall motion score (all P<0.05). The treatment significantly decreased serum ACE activities (P<0.01) and increased plasma bradykinin concentrations (P<0.05). The extent of reduction of left ventricular wall motion score by perindopril was closely correlated with that of inhibition of serum ACE activities (P<0.01) and with that of increase in plasma bradykinin concentrations (P<0.05). By contrast, no such beneficial changes were noted in the control group. These results provide the first evidence that long-term treatment with perindopril exerts anti-ischemic effects on the myocardial ischemia induced by increased myocardial oxygen demand in patients with CAD. PMID- 11855669 TI - Characterization of beta-adrenoceptor subtype in bladder smooth muscle in cynomolgus monkey. AB - We first investigated the relaxations of the urinary bladder induced by beta adrenoceptor agonists in anesthetized cynomolgus monkeys and then employed a variety of beta-adrenoceptor agonists and antagonists in vitro to identify the beta-adrenoceptor subtype responsible for the relaxation (using isolated monkey detrusors). Isoprenaline reduced bladder pressure in a dose-dependent manner. Isoprenaline, noradrenaline and adrenaline each produced a concentration dependent relaxation of isolated detrusor strips, the rank order of relaxing potencies being isoprenaline > noradrenaline > adrenaline. Subtype-selective beta adrenoceptor agonists also relaxed isolated detrusor strips, the rank order of potencies being CGP-12177 > BRL 37344 > dobutamine, salbutamol, procaterol > xamoterol. In the antagonist experiment, bupranolol (beta-antagonist, 10(-6) to 10(-5) M) and SR 58894A (beta3-antagonist, 10(-7) to 10(-5) M) caused a rightward shift of the concentration-relaxation curve for isoprenaline, but CGP-20712A (beta1-antagonist, 10(-9) to 10(-7) M) and ICI-118551 (beta2-antagonist, 10(-9) to 10(-7) M) did not. The present functional study provides the first evidence that relaxation of the monkey detrusor by beta-adrenoceptor activation is mediated via the beta3-subtype. PMID- 11855670 TI - Modification of 5-hydroxytryptophan-evoked 5-hydroxytryptamine formation of guinea pig colonic mucosa by reactive oxygen species. AB - We studied whether reactive oxygen species (ROS) generated by normal colonic mucosa affect 5-hydroxytryptophan (5-HTP)-evoked 5-HT formation (measured as the sum of 5-HT plus 5-hydroxyindole acetic acid (5-HIAA) accumulation) of guinea pig's isolated colonic mucosa. Catalase (3000-6000 U/ml), a hydrogen peroxide (H2O2) scavenger or diphenylene iodonium (DPI, 10-100 microM), an NADPH oxidase inhibitor, concentration-dependently caused an increase of the sum of 5-HT plus 5 HIAA accumulation in the presence of 5-HTP (10 microM), but these drugs did not significantly affect the 5-HT-metabolite in the colonic mucosa measured as the ratio of 5-HIAA/5-HT. Exogenously applied H2O2 (10-100 microM) concentration dependently inhibited the sum of 5-HT plus 5-HIAA accumulation. In contrast, neither superoxide dismutase (SOD, 100-300 U/ml), superoxide anion scavenger, nor dimetyl sulfoxide (1-5%, DMSO), a hydroxyl radical scavenger affected the sum of 5-HT plus 5-HIAA accumulation. Moreover, mucosa ROS generation was estimated using the chemiluminescence technique. SOD (100-300 U/ml), catalase (3000-6000 U/ml) or DPI (10-100 microM), concentration-dependently reduced luminol-enhanced chemiluminescence signal from the colonic mucosa, while allopurinol (10-100 microM), a xanthine oxidase inhibitor, did not affect the chemiluminescence signal. These results suggest that ROS is formed through an NADPH oxidase system in the guinea pig colonic mucosa, where it exerts a modulatory effect on mucosal 5-HT formation upon addition of 5-HTP. Thus, ROS formation from normal colonic mucosa could be considered to contribute to the control of 5-HT production in mucosa enterochromaffin cells. PMID- 11855671 TI - Effect of oral OPC-28326, a selective femoral arterial vasodilator, on hindlimb subcutaneous tissue temperature in conscious dogs under buprenorphine sedation. AB - In dogs, rectal temperature was decreased to about 36 degrees C at 2 to 2.5 h after sedation with buprenorphine (0.3 mg/body, i.m.), and hindlimb subcutaneous tissue temperature (T(SC)) in the thigh decreased in a similar manner. T(SC) in the dorsum of the foot showed a greater decrease than that in the thigh. OPC 28326 (4-(N-methyl-2-phenylethylamino)-1-(3,5-dimethyl-4-propionyl-aminobenzoyl) piperidine hydrochloride monohydrate) at doses of 0.3, 1.0 and 3.0 mg/kg, p.o. inhibited the buprenorphine-induced decrease in T(SC) in the dorsum dose dependently, but had little effect on that in the thigh or rectal temperature. In conclusion, T(SC) in the extremities were more sensitive to core temperature and peripheral circulation. PMID- 11855672 TI - Gene expression of enzymes comprising the polyol pathway in various rat tissues determined by the competitive RT-PCR method. AB - The quantitative measurements of aldose reductase (AR) and sorbitol dehydrogenase (SDH) gene expression in various rat tissues were performed by the competitive reverse transcription-polymerase chain reaction (RT-PCR). AR mRNA was detectable in all tissues analyzed with pronounced differences in the amounts. SDH mRNA was most abundant in testes and liver, but was absent from lens. The estimation of the AR cDNA/SDH cDNA ratio showed that the relative abundance of SDH to AR differs among tissues. These results indicate that different tissues contain varying amounts of AR mRNA and SDH mRNA; that is, each tissue has its own polyol pathway activity. PMID- 11855673 TI - Sympathetic cholinergic vasodilation of skeletal muscle small arteries. AB - Recently we have studied the direct vasomotor response of the hindlimb extramuscular large arteries (internal diameter, 500-1400 microm) and intramuscular small arteries (internal diameter, 50-500 microm) of in vivo thick skeletal muscle during activation of sympathetic cholinergic nerve in anesthetized cats. The hypothalamic defense area was electrically stimulated so as to induce a profound increase in femoral blood flow mediated by sympathetic cholinergic fibers. To visualize the vascular arrangement from the extramuscular large feeding arteries to small arteries in the triceps surae muscle, we developed a new X-ray TV system. The internal diameter, flow velocity, and volume flow of arterial blood vessels were directly measured before and during stimulation of the hypothalamic defense area. The major new finding is that the hypothalamic stimulation causes an intense increase in the internal diameter of small arteries in skeletal muscle, which is abolished either by cholinergic blockade or by the section of the sciatic nerve, but not by combined alpha- and beta-adrenergic blockade. In contrast, the internal diameter of the extramuscular larger arteries does not change during the hypothalamic stimulation, but their flow velocity and volume flow increase. These findings indicate that sympathetic cholinergic vasodilation occurs at intramuscular small arteries with internal diameter of 50-500 microm, which in turn increases flow velocity and volume flow of upstream blood vessels. PMID- 11855674 TI - Purinergic modulation of vascular sympathetic neurotransmission. AB - It is generally agreed that the release of norepinephrine (NE) is inhibited by activation of prejunctional purinoceptor. We examined the pharmacological properties of purinoceptors on vascular sympathetic nerve terminals and the source of endogenous adenyl purines. Electrically (1 Hz) evoked NE-release was inhibited by not only P1-agonists but also P2-agonists. Although the inhibition induced by P2-agonists was blocked by P1-antagonists, P2-agonists-induced inhibition was not due to the breakdown to adenosine. Therefore, there may be a new class of purinoceptor that is activated by both P1- and P2-agonists and antagonized by P1-antagonists. Electrical stimulation at 8 Hz but not at 1 Hz evoked the release of adenyl purines such as ATP, ADP, AMP and adenosine, in addition to NE; and the purines-release was blocked by an alpha1-antagonist. Methoxamine, an alpha1-agonist, also evoked the release of purines. Electrically (1 Hz)-evoked NE-release was inhibited by methoxamine, and this inhibition was blocked by not only an alpha1-antagonist but also a P1-antagonist. Therefore, the activation of alpha1-adrenoceptor appeared to release purines, which in turn inhibited NE-release via prejunctional purinoceptors. From these results, it is suggested that the unique purinoceptor and the endogenous purines released from alpha1-adrenoceptor-sensitive sources participate in the antidromic transsynaptic modulation of vascular sympathetic neurotransmission. PMID- 11855675 TI - Neurogenic cerebral vasodilation mediated by nitric oxide. AB - In cerebral arteries isolated from most of mammals, nerve stimulation produces relaxations in contrast to contractions in peripheral arteries. The relaxant mechanism is found to be non-adrenergic and non-cholinergic, but the neurotransmitter is not clarified until recently. Based on several functional and histological studies with isolated cerebral arteries, nitric oxide (NO) is now considered to be a neurotransmitter of the vasodilator nerve and the nerve has been called a nitroxidergic (nitrergic) nerve. Upon neural excitation, calcium influxed through N-type Ca2+ channels activates neuronal NO synthase, and then NO is produced by the enzyme from L-arginine. The released NO activates soluble guanylate cyclase in smooth muscle cells, resulting in relaxation with a cyclic GMP-dependent mechanism. The functional role and neuronal pathway have also been investigated in anesthetized dogs and Japanese monkeys. The nitroxidergic (nitrergic) nerves innervating the circulus arteriosus, including the anterior and middle cerebral and posterior communicating arteries, are found to be postganglionic nerves originated from the ipsilateral pterygopalatine ganglion and tonically dilate cerebral arteries in the resting condition. Our findings suggest that the nitroxidergic (nitrergic) nerve plays a physiologically important role to maintain a steady blood supply to the brain. PMID- 11855676 TI - Regulation of vascular function by perivascular calcitonin gene-related peptide containing nerves. AB - The rat mesenteric artery is innervated by nonadrenergic noncholinergic (NANC) vasodilator nerves in which calcitonin gene-related peptide (CGRP), a potent vasodilator peptide, acts as a vasodilator transmitter. The inhibition of CGRPergic nerve function potentiates a vasoconstrictor response mediated by the sympathetic adrenergic nerve, suggesting that CGRPergic nerves inhibit adrenergic function and play a role in the regulation of mesenteric vascular tone. In contrast, norepinephrine released from adrenergic nerves presynaptically inhibits neurotransmission of CGRPergic nerves. Thus, both nerves reciprocally control the vascular tone. Pathophysiological studies have shown that an age-related decrease in CGRPergic nerve-mediated vasodilation, neurogenic CGRP release and CGRP mRNA levels in the dorsal root ganglia are found in spontaneously hypertensive rats (SHR), indicating a reduced function of CGRPergic nerves. Long-term treatment with angiotensin converting enzyme inhibitor and angiotensin II-receptor antagonist restores the reduced function of CGRPergic nerves, suggesting the involvement of angiotensin II in the malfunction of CGRPergic nerves in SHR. PMID- 11855677 TI - Protective effect of thiaton, an antispasmodic drug, against indomethacin-induced intestinal damage in rats. AB - The effect of thiaton [3-(di-2-thienylmethylene)-5-methyl-trans-quinolizidinium bromide], an antispasmodic drug, on indomethacin-induced intestinal damage was examined in rats. The animals were given indomethacin, s.c., and the intestinal mucosa was examined 24 h later. Thiaton or atropine was given s.c. twice, 30 min before and 8 h following indomethacin. Indomethacin caused intestinal damage, accompanied with increase in enterobacterial translocation as well as inducible nitric oxide synthase (iNOS) and myeloperoxidase (MPO) activities, and these changes were significantly prevented by supplementation with 16,16-dimethyl prostaglandin E2 (dmPGE2). Treatment of the animals with thiaton dose-dependently prevented the intestinal damage, together with the suppression of MPO and iNOS activities, and these effects were similarly observed by atropine. The increase of bacterial translocation was also significantly prevented by both thiaton and atropine, similar to dmPGE2. Indomethacin enhanced intestinal motility, and this effect was inhibited by either thiaton, atropine or dmPGE2. The intestinal mucus and fluid secretions were decreased by indomethacin but enhanced by dmPGE2. Both thiaton and atropine slightly decreased these secretions under basal conditions but significantly reversed the decrease in the secretions caused by indomethacin. These results suggest that thiaton protects the small intestine against indomethacin-induced damage and inflammatory changes, and this effect is related with prevention of enterobacterial translocation, the process being associated with inhibition of intestinal hypermotility caused by indomethacin, probably due to anti-muscarinic action. PMID- 11855678 TI - Effects of vitamin K2 (menatetrenone) on calcium balance in ovariectomized rats. AB - Vitamin K2 (menatetrenone) has been used for the treatment of osteoporosis in Japan. We investigated the effects of ovariectomy (OVX) and vitamin K2 on the calcium (Ca) balance in 20-week-old female Fischer rats. Vitamin K2 (31 mg/kg per day) was given to animals as a dietary supplement. At weeks 4 and 8 after OVX, a Ca balance study was performed for 5 days. The intestinal Ca transport was determined using the everted gut-sac technique at week 9. The Ca balance was poorer in the OVX-control group than in the sham-control group at weeks 4 and 8 after OVX. The Ca balance improved significantly in the vitamin K2 groups as compared with the sham- and OVX-control groups. The intestinal Ca transport decreased due to OVX and was higher in the vitamin K2 administration groups than in the sham- and OVX-control groups, but not to a significant extent. The bone mineral density in the femoral metaphysis as well as the cortical area and cortical thickness in the femoral diaphysis in the OVX-control group were lower than in the sham-control group. The administration of vitamin K2 significantly inhibited an OVX-induced decrease in cortical area and cortical thickness in the femur. These findings suggest that the poor Ca balance observed in ovariectomized rats may be improved by vitamin K2; vitamin K2 may be involved in preventing bone loss in vivo. PMID- 11855679 TI - Lung tissue distribution after intravenous administration of grepafloxacin: comparative study with levofloxacin. AB - The aim of the present study is to study the pharmacokinetics in plasma, lung lymph and bronchial washing fluid after intravenous infusion of grepafloxacin (GPFX), in comparison with those of levofloxacin (LVFX). Four conscious sheep with chronically instrumented lung lymph fistulas and tracheotomy were prepared. GPFX and LVFX concentrations in plasma and lung lymph after intravenous infusion of the drugs (10 mg/kg) for over 10 min were measured. In addition serial bronchial washing with 50 mL normal saline was performed to obtain epithelial lining fluid (ELF) at 2, 4, 6, 8, 12, 24 h after the intravenous administration. The time courses of lung lymph concentration were almost identical to those of the concomitant levels of both GPFX and LVFX in plasma, suggesting that both GPFX and LVFX could be easily moved from plasma to pulmonary interstitium and/or lung lymph circulation. However, GPFX concentrations of ELF were significantly higher than LVFX concentrations over time after the administration. In addition, intracellular concentrations in ELF of GPFX were also extremely high compared with those of LVFX. These results demonstrated that penetration of GPFX in bronchial wall, bronchial epithelium and/or phagocytic cells was superior to that of LVFX. These observations suggest that the pharmacokinetic characteristics of GPFX in the lung may provide a new insight into the strategy for clinical treatment of various pulmonary infections, especially cytotropic bacterial infections. PMID- 11855680 TI - Human organic anion transporters mediate the transport of tetracycline. AB - The purpose of this study was to elucidate the molecular mechanism for renal tetracycline transport by human organic anion transporters (hOATs) using proximal tubular cells stably expressing hOATs. The cells stably expressing hOAT1, hOAT2, hOAT3 and hOAT4 exhibited a higher amount of [3H]tetracycline uptake compared with mock cells. The apparent Km values for hOAT2-, hOAT3- and hOAT4-mediated tetracycline uptakes were 439.9 +/- 23.0, 566.2 +/- 28.4 and 122.7 +/- 16.0 microM, respectively. Tetracycline significantly inhibited the organic anion uptake by hOAT1, hOAT2 and hOAT4, but not hOAT3. In addition, oxytetracycline, minocycline and doxycycline inhibited the organic anion uptake by hOAT1, whereas oxytetracycline, minocycline but not doxycycline inhibited the organic anion uptake by hOAT2. In contrast, oxytetracycline, minocycline and doxycycline exhibited no significant inhibitory effects on the organic anion uptake by hOAT3 and hOAT4. HOAT1 and hOAT4 mediated the efflux of tetracycline, but hOAT2 and hOAT3 did not. These results suggest that hOAT1, hOAT2 and hOAT3 mediate the basolateral uptake and/or efflux of tetracycline, whereas hOAT4 is responsible for the reabsorption as well as the efflux of tetracycline in the apical side of the proximal tubule. These pharmacological characteristics of hOATs may be significantly related to events associated with the development of tetracycline induced nephrotoxicity in the human kidney. PMID- 11855681 TI - Genomic structure of the rat and mouse histamine N-methyltransferase gene. AB - Northern blotting analysis revealed different tissue distribution patterns of HMT mRNA between mice and rats. In the mouse, mRNA expression was detected in the brain, kidney and liver. In the rat, there was an extremely high mRNA signal only in the kidney. We isolated and characterized the rat and mouse histamine N methyltransferase (HMT) genes from genomic DNA libraries. The rat HMT gene consists of 6 exons and 5 introns. The mouse HMT gene structure was similar to that of the rat, but had one additional exon 5' upstream from the exon containing a start codon, resulting in seven exons. Several long interspersed repetitive elements were located in the 5' flanking region of the rat and mouse HMT gene. Despite high sequence conservation of the regions around exon 6 and the 3' flanking region, the 5' flanking region had little similarity between the rat and mouse. Marked sequence similarities between rat and mouse introns were present near splice sites and outside the junction residues, suggesting the evolutionary relationship between the structural features of the rat and mouse HMT genes. This observation may explain the species difference of the tissue expression pattern of HMT mRNA. PMID- 11855682 TI - Role of perivascular sympathetic nerves and regional differences in the features of sympathetic innervation of the vascular system. AB - Maintenance of blood pressure is mostly dependent on sympathetic "tone", and the sympathetic nerve innervates the entire vascular bed, excepting the capillaries. Although norepinephrine (NE) is the principal neurotransmitter released upon sympathetic nerve stimulation, neuropeptide Y and ATP are cotransmitters in various vascular tissues. In addition, dopamine and epinephrine, as well as acetylcholine, have been shown to be sympathetic neurotransmitters in specific vasculatures. Transmitter NE release is modified by a number of endogenous substances including the transmitter itself. Chronic denervation of the preganglionic fiber induces an increase in NE release per pulse, indicating postganglionic neuronal supersensitivity. So far, three main adrenoceptor types have been shown, alpha1, alpha2 and beta, each of which is further divided into at least three subtypes, as well as the alpha1L-adrenoceptor, a phenotype of the cloned alpha1a-adrenoceptor, in the blood vessel. Thus, the response of vessels with different receptor types to a transmitter varies quantitatively and even qualitatively from one vessel to another. The remarkable diversity in the sympathetic innervation mechanism in the vascular system may play an important role in regional variations in the regulation of blood flow. The sympathetic nerve also exerts long-term trophic action on the blood vessel. In conclusion, the sympathetic nervous system plays an important role not only in the regulation of cardiovascular dynamics but in the maintenance of the vessel structure, as well. PMID- 11855683 TI - Inhibitory effects of pre-ischemic and post-ischemic treatment with FR 168888, a Na+/H+ exchange inhibitor, on reperfusion-induced ventricular arrhythmias in anesthetized rat. AB - Effects of pre-ischemic and post-ischemic treatment with FR 168888 (5 hydroxymethyl-3-(pyrrol-1-yl) benzoylguanidine methanesulfonate), a Na+/H+ exchange inhibitor, on reperfusion-induced ventricular arrhythmias were examined in an ischemia/reperfusion model of anesthetized rat. FR 168888 (0.3 mg/kg) significantly reduced the incidence of ventricular fibrillation (VF) and mortality induced by reperfusion following 5-min coronary occlusion, when it was intravenously administered 5 min before coronary artery occlusion. Post-ischemic treatment with FR 168888 (0.3-10 mg/kg), i.e. given 3 min after the start of occlusion, reduced the incidence of VF and mortality. In order to examine the optimal time of administration, FR 168888 (3 mg/kg) was administered 1 or 3 min after the start of occlusion or immediately before reperfusion. There was no significant difference in the reduction of VF and mortality among the three post ischemic treatment groups. FR 168888 (3 and 10 mg/kg) significantly increased the blood pressure during ischemia without affecting the heart rate. These results indicate that FR 168888 has antiarrhythmic effects on reperfusion-induced arrhythmias even administered after coronary occlusion. PMID- 11855684 TI - Effects of growth hormone and insulin-like growth factor 1 deficiency on ageing and longevity. AB - Present knowledge on the effects of growth hormone (GH)/insulin-like growth hormone (IGF)1 deficiency on ageing and lifespan are reviewed. Evidence is presented that isolated GH deficiency (IGHD), multiple pituitary hormone deficiencies (MPHD) including GH, as well as primary IGE1 deficiency (GH resistance, Laron syndrome) present signs of early ageing such as thin and wrinkled skin, obesity, hyperglycemia and osteoporosis. These changes do not seem to affect the lifespan, as patients reach old age. Animal models of genetic MPHD (Ames and Snell mice) and GH receptor knockout mice (primary IGF1 deficiency) also have a statistically significant higher longevity compared to normal controls. On the contrary, mice transgenic for GH and acromegalic patients secreting large amounts of GH have premature death. In conclusion longstanding GH/IGF1 deficiency affects several parameters of the ageing process without impairing lifespan, and as shown in animal models prolongs longevity. In contrast high GH/IGF1 levels accelerate death. PMID- 11855685 TI - The role of insulin-like growth factor 1 and insulin in ageing and atherosclerosis. AB - With advancing age insulin-like growth factor (IGF)1 blood levels decrease continuously, but with great interindividual differences. There is a relationship between the IGF1 serum concentration and biomarker behaviour, indicating that growth hormone (GH) secretion is a determinant of organismic well being and surviving in advanced age. In contrast, the secretion of insulin rises with age, which is related to both increasing body fat mass and ageing itself. In vitro insulin stimulates the proliferation, migration and collagen secretion of human vascular smooth muscle cells (SMCs). The mechanism underlying these processes mainly involves occupancy of ICF1 receptors by insulin, with the exception of migration. With advancing age of the donor, the in vitro proliferation rate and migration capacity of SMCs decreases. When insulin or IGF1 is added, there is no reversibility, so that there is no recovery to the values of SMCs from young donors. The blockade of Ca2+ channels by diltiazem inhibits the in vitro stimulation by IGF1 and insulin on SMC proliferation and migration. We conclude that the acceleration of ageing is related to the decline of IGF1 in such a manner that ageing rates progress as GH secretion diminishes. Biomarkers are affected correspondingly. The role of insulin in atherogenesis is related to hyperinsulinaemia, but the increase in insulin secretion belongs to the process of ageing regulation. Nevertheless, the effect of insulin in changing the phenotype of SMCs is atherogenic. Diltiazem may thcrefore act as an antiatherogenic agent. In advanced age the risk of atherogenesis decreases because of lowered propensity of SMCs to proliferate and migrate, which is probably due to a greater proportion of senescent cells. PMID- 11855687 TI - The ageing female reproductive axis II: ovulatory changes with perimenopause. AB - Perimenopause, a complex physiological transition for midlife women, begins with changes in experiences many years before cycles become irregular, oestradiol levels decrease or follicle-stimulating hormone levels increase. Erratic and average higher oestradiol levels as well as shorter luteal phase lengths and lower progesterone levels occur during perimenopause. These ovarian changes may be causally related to lower inhibin production but the dynamic prospective inter relationships within women are not well documented. This review will first define perimenopause and then explore the limited published data on ovulatory characteristics in perimenopause. In addition, it will report preliminary prospective observational data on menstrual cycles and ovulation in initially ovulatory women followed through the perimenopause. Prospective data suggest that ovulation disturbances begin early in perimenopause and increase with irregular cycles. Combined with higher oestradiol levels they may cause menorrhagia. It is not yet known whether disturbances of ovulation relate to bone loss in perimenopausal, as in premenopausal, women. It is also not known whether progesterone therapy can effectively counteract the end organ (breast, endometrial, brain) effects of higher/erratic oestradiol levels and effectively treat perimenopausal vasomotor and other symptoms. PMID- 11855686 TI - The ageing female reproductive axis I. AB - The female reproductive axis includes the hypothalamo-pituitary unit, the ovaries and the uterus. While changes in the brain may contribute to reproductive ageing, the major focus of current research is on the ovary, where the progressive loss of follicles ultimately leads to absent follicular function and consequent permanent cessation of menstruation, the menopause. The pituitary gonadotropins, follicle-stimulating hormone (FSH) and luteinizing hormone, stimulate ovarian secretion of oestradiol and the inhibins from follicular granulosa cells, and androgens from interstitial cells, including the theca. A primary event in the ageing of the reproductive axis appears to be a decline in the secretion of inhibin B as follicle numbers fall. This leads to a slow rise in FSH in women who continue to cycle regularly, particularly in the last decade of reproductive life. As the menopause approaches, decreasing concentrations of both oestradiol and inhibin B lead to more marked increases in the gonadotropins, which reach their postmenopausal peak 2-3 years after final menses. In contrast, total testosterone concentrations are maintained across the menopausal transition, with a fall in sex hormone binding globulin (SHBG) and hence a rise in free testosterone. PMID- 11855688 TI - The thyroid axis in ageing. AB - The hypothalmo-pituitary thyroid axis, among various endocrine systems, undergoes physiological alterations associated with the ageing process. Directly age related changes have to be distinguished from indirect modifications which are caused by simultaneous thyroidal or non-thyroidal illness or other physiological or pathophysiological states whose incidence increases with age. In summary, direct changes of the hypothalmo-pituitary-thyroid axis seem to be subtle and suggestive of a decreased hypothalamic stimulation of thyroid function. In parallel, disease-specific alterations such as the development of thyroid autonomy or changes in energy intake or sleep lead to pronounced alterations of thyroid function with age which may dominate the underlying ageing of the hypothalmo-pituitary thyroid axis itself. The following article attempts to delineate some aspects of the interplay of the regulation of thyroid function and the ageing process. PMID- 11855689 TI - Critical illness as a model of hypothalamic ageing. AB - In this review we shall consider the endocrine changes seen in the various hypothalamic pituitary-target gland axes at different stages of critical illness and conclude by comparing these changes with those seen in the normal ageing process. PMID- 11855690 TI - Glucose tolerance, glucose utilization and insulin secretion in ageing. AB - Ageing is associated with an increased incidence of hypertension, macrovascular disease and type 2 diabetes (non-insulin-dependent diabetes). It has been suggested that a common mechanism may be responsible for all of these pathological states since all of these conditions often cluster in the same individual. Epidemiological and clinical data have consistently demonstrated an association between insulin resistance and/or hyperinsulinaemia and glucose intolerance, dyslipidaemia and elevated systolic blood pressures. Therefore, insulin resistance and hyperinsulinaemia have been proposed as the causal link among the elements of the clusters. The elderly are more glucose intolerant and insulin resistant, but it remains controversial whether this decrease in function is due to an inevitable consequence of 'biological ageing' or due to environmental or lifestyle variables, noticeably increased adiposity/altered fat distribution and physical inactivity. An increase of these modifiable factors has been shown to result in increases in insulin resistance and hyperinsulinaemia and vice versa. However, insulin secretion appears to decrease with age even after adjustments for differences in adiposity, fat distribution and physical activity. The glucose intolerance of ageing may be due, in part, to decreased insulin sensitivity of pancreatic / cells to insulinotropic gut hormones (GLP1/GIP) and in part to alterations of hepatic glucose production. PMID- 11855691 TI - Endocrine causes of age-related bone loss and osteoporosis. AB - Women have an early postmenopausal phase of rapid bone loss that lasts for 5-10 years after menopause, whereas both ageing women and men have a slow continuous phase of bone loss that lasts indefinitely. In women, the rapid phase is mediated mainly by loss of the direct restraining effect of oestrogen on bone cell function, whereas the slow phase is mediated mainly by the loss of oestrogen action on extraskeletal calcium homeostasis leading to net calcium wasting and secondary hyperparathyroidism. Because elderly men have low serum bioavailable oestrogen and testosterone levels, and because recent data suggest that oestrogen is the main sex steroid regulating bone metabolism in men, oestrogen deficiency may also be the principal cause of bone loss in elderly men. Decreased bone formation contributes to bone loss in both genders and may be caused by a decreased production of growth hormone and IGF1 as well as oestrogen and testosterone deficiency. Other changes in endocrine secretion, although present in the elderly, seem less important in the pathophysiology of age-related bone loss and osteoporosis. PMID- 11855692 TI - Ageing, stress and the brain. AB - Ageing of the brain is an important factor in overall ageing and mortality, and new insights have clarified the relationship between neuroregulation and ageing. First, neuronal loss in normal ageing is now known to be a minor change. Loss of synapses through dystrophic neuronal change is the hallmark of normal ageing. Second, similar dystrophic changes occur in the brain with chronic stress. In both instances, forebrain sites experience loss of synaptic input from brainstem regulatory nuclei. Third, functional ageing is attributed in part to lifetime stress, under the concept of 'allostatic load'. Being inseparable from the functions of appraising and responding to stress, the brain is an ultimate mediator of stress-related mortality, through hormonal changes that lead to proximate pathologies like hypertension, glucose intolerance, cardiovascular disease and immunological impairment. In chronic stress the brain shows clear allostatic compensations that lead to pathology. Two subtle and chronic mechanisms that may mediate brain pathology and accelerated ageing in chronic stress are proposed. These are abnormal glucocorticoid receptor (GR) occupancy over the 24 h cycle, and elevated body temperature. These factors lead to GR mediated tissue changes and to acceleration of general cellular ageing mechanisms. Human depression is discussed as an exemplary demonstration of these principles. PMID- 11855693 TI - Ageing and water homeostasis. AB - This review outlines current knowledge concerning fluid intake and volume homeostasis in ageing. The physiology of vasopressin is summarized. Studies have been carried out to determine orthostatic changes in plasma volume and to assess the effect of water ingestion in normal subjects, elderly subjects, and patients with dysautonomias. About 14% of plasma volume shifts out of the vasculature within 30 minutes of upright posture. Oral ingestion of water raises blood pressure in individuals with impaired autonomic reflexes and is an important source of noise in blood pressure trials in the elderly. On the average, oral ingestion of 16 ounces (473ml) of water raises blood pressure 11 mmHg in elderly normal subjects. In patients with autonomic impairment, such as multiple system atrophy, strikingly exaggerated pressor effects of water have been seen with blood pressure elevations greater than 75 mmHg not at all uncommon. Ingestion of water is a major determinant of blood pressure in the elderly population. Volume homeostasis is importantly affected by posture and large changes in plasma volume may occur within 30 minutes when upright posture is assumed. PMID- 11855694 TI - Alterations in the ageing corticotropic stress-response axis. AB - The problem with determining whether ageing is followed by perturbations of the regulation of the hypothalamic pituitary adrenal (HPA) axis is that ageing per se is very difficult to separate from the effects of environmental insults. Data in ageing rodents indicate that with age the winding-down of cortisol after challenges is prolonged. This is probably due to an insufficient feedback regulation by glucocorticoid receptors in specific fields in the hippocampus. The functional and topographic characteristics of these changes are identical to those seen after prolonged stress, suggesting that such factors might be of significance. Data in humans also suggest that with age basal cortisol secretion, diurnal rhythm, and the early response to challenges are not affected but similar to animal data the return to baseline values after stimulation seems to be delayed, probably due to a diminished feedback control. Several studies suggest that common diseases of age, for example cardiovascular disease and type 2 diabetes mellitus, are associated with similar HPA axis perturbations as those seen in old subjects. Recent population studies indicate that adrenal hyperactivity, associated with a stressful environment, is generating risk factors for these diseases. This is likely to be dependent on genetic susceptibility, and associated polymorphisms have been found in several candidate genes of importance for neuroendocrine and autonomic regulations. PMID- 11855695 TI - Male reproductive ageing: human fertility, androgens and hormone dependent disease. AB - The waning of male virility with age, in all its ambiguities, has always intrigued humans, prompting innumerable approaches to staving it off. In modern terms, advancing age impacts on all aspects of male reproductive health sexuality, fertility and androgenization with differing extents and tempo. With ageing, male sexual function declines predominantly due to vasculogenic defects in cavernosal haemodynamics, whereas libido and ejaculation are less affected. This raises the potential for prevention and treatment of erectile dysfunction as an early clinical manifestation of atherosclerosis. After maturity, male fertility persists throughout life but decreases modestly with age presumably due to concomitant decline in sexual activity rather than in sperm output or function, although systematic population studies of the latter are difficult. Male ageing is associated with a progressive, partial and variable degree of androgen deficiency, but the clinical and public health significance remain to be established. Available evidence suggests that androgen supplementation is unlikely to prolong life expectancy but might improve quality of life through prevention of apparently age-related declines in androgen-sensitive tissues. The appropriate target population, treatment modality and objectives remain to be established by further controlled clinical studies. PMID- 11855696 TI - Male reproductive ageing: using the brown Norway rat as a model for man. AB - The Brown Norway (BN) rat is an excellent model for male reproductive ageing. We and others have shown that with ageing, the BN rat exhibits low serum testosterone, low Leydig cell steroidogenic capacity, decreased Sertoli cell function and number, marked reduction in seminiferous tubule volume and sperm content, and accelerated germ cell apoptosis. These testicular changes are the result of a combination of a primary testicular defect and a secondary hypothalamic dysfunction. Leydig cell dysfunction results from decreased activities of the steroidogenic enzymes and Leydig cell secretory capacity and is not corrected by daily administration of replacement luteinizing hormone (LH), suggesting a primary testicular defect. However ageing in male BN rats is associated with decreased LH pulse amplitude, reduced gonadotropin releasing hormone (GnRH) and gonadotropin responsiveness to excitatory amino acids, and decreased GnRH mRNA and peptide in the hypothalamus. We have further shown in the hypothalamus of ageing BN rats that while the excitatory amino acid receptor content is reduced, nitric oxide synthase (NOS) activity is increased which is due to increased inducible (iNOS) but not neuronal NOS (nNOS). The increased iNOS protein in the hypothalamus is associated with increased peroxynitrite formation and neuronal cell apoptosis. We conclude that increased hypothalamic levels of iNOS may result in neurotoxicity in the hypothalamus leading to loss of hypothalamic GnRH secretory cells and impaired GnRH pulsatile secretion that contributes to the abnormal Leydig cell function characteristic of male reproductive ageing. PMID- 11855697 TI - Mechanisms of conjoint failure of the somatotropic and gonadal axes in ageing men. AB - Endogenous growth hormone (GH) production falls by 50% every 7 years and bioavailable testosterone concentrations decline concomitantly by 12-15% every decade in ageing men. Despite this temporal parallelism, the neuroendocrine bases of the somatopause and gonadopause are not known. This knowledge deficit contrasts with the recent unfolding of new insights into the nature of oestrogen dependent control of the GH-insulin-like growth factor (IGF)1 axis in pre- and postmenopausal women. The present overview examines the postulate that the pathophysiology of somatopause and gonadopause in ageing men is bidirectionally linked. According to this broader thesis, hyposomatotropism accentuates Leydig cell steroidogenic failure and, conversely, progressive androgen deficiency exacerbates the decline in GH-IGF1 output in ageing. PMID- 11855698 TI - Rising to the challenge of the new NCEP ATP III guidelines: exceeding current therapeutic limitations. AB - The third set of guidelines recently issued by the National Cholesterol Education Program (NCEP) differs from the second set issued in 1993 in several ways. The third set introduced a quantitative risk scoring system and identified and/or reclassified certain groups of patients at high or moderate risk for a coronary event. Among these groups are patients with type 2 diabetes, and patients with multiple risk factors other than coronary heart disease or diabetes that cumulatively confer high risk for a coronary event. However, the new guidelines also present physicians with the major challenges of identifying these patients, determining their true risk, and implementing the recommended approaches to treatment in clinical practice settings. Although reducing elevated levels of low density lipoprotein (LDL) cholesterol remains the primary focus of therapy, the new NCEP guidelines also include strategies to identify and treat patients with low levels of high-density lipoprotein (HDL) cholesterol and/or elevated triglyceride levels. Just as there is "good" cholesterol (HDL) and "bad" cholesterol (LDL), there are also "good" triglycerides, which contain high concentrations of triglyceride remnants and are associated with low risk, and "bad" triglycerides, which contain high concentrations of cholesterol remnants and are associated with increased risk. The mechanisms by which "bad" triglycerides develop explain why elevated triglycerides and low HDL--and patients with the metabolic syndrome--warrant special attention. These mechanisms and others also suggest new targets for therapeutic intervention and the development of new drugs that will correct lipid and lipoprotein abnormalities through a number of different metabolic pathways. PMID- 11855699 TI - State of the art in cholesterol management: targeting multiple pathways. AB - The relationship between increased serum levels of total cholesterol and increased risk of coronary heart disease (CHD) is widely accepted, as is the corollary that lowering elevated levels of low-density lipoprotein (LDL) cholesterol, a significant risk factor, reduces CHD mortality. Although statins are the most widely used agents to lower LDL cholesterol levels and demonstrate significant clinical benefits, alternate and/or additional treatment strategies are also being studied. One such approach for which several classes of agents are available is the nonspecific inhibition of intestinal cholesterol and bile acid absorption. Although all of these inhibitors-which include bile acid sequestrants, cholesterol absorption blockers, and plant stanols-reduce cholesterol concentrations within liver cells and increase the expression of LDL receptors, they are limited in their effectiveness because of poor tolerability and compensatory effects by the liver that blunt their ability to lower LDL. Ideally, the combination of a statin and another agent that specifically targets cholesterol absorption and promotes excretion while simultaneously decreasing synthesis may provide the greatest therapeutic benefit. The recent development of selective cholesterol inhibitors, including ezetimibe, provides an approach to lowering LDL that has significant potential both as monotherapy and in combination therapy. PMID- 11855700 TI - Preclinical and clinical pharmacology of a new class of lipid management agents. AB - Ezetimibe, the first in a new class of selective cholesterol absorption inhibitors, reduces plasma levels of low-density lipoprotein (LDL) cholesterol by selectively preventing the intestinal uptake of dietary and biliary cholesterol. Initial studies in animals show that ezetimibe lowers both serum and liver cholesterol in a dose-dependent manner without decreasing the absorption of triglycerides or fat-soluble vitamins. It also appears to function primarily as its glucuronidated derivative and, in this form, localizes almost exclusively to the intestinal mucosal cells after initial absorption and glucuronidation in the gut and liver. Preclinical studies show that ezetimibe also lowers LDL effectively, whereas studies in apolipoprotein E knockout mice show that it reduces LDL, very-low-density lipoproteins, and the formation of atherosclerotic plaque in the aorta. Because of its safety and specificity, and because its mechanism of action complements that of the statins, ezetimibe is an attractive component in combination therapy. Coadministration of ezetimibe and a statin in dogs demonstrates larger reductions in cholesterol levels than therapy with either agent alone. PMID- 11855701 TI - An investigative look: selective cholesterol absorption inhibitors--embarking on a new standard of care. AB - Phase 2 and 3 clinical trials evaluating the selective cholesterol absorption inhibitor ezetimibe have demonstrated that the drug is safe and effective, both as monotherapy and in combination with several statins. Placebo-controlled phase 2 studies of 8 and 12 weeks' duration established that ezetimibe monotherapy achieved maximum cholesterol lowering at doses between 10 and 20 mg daily. Additional dose-scheduling studies demonstrated that evening dosing was only slightly more effective than morning dosing, and that the drug could be taken with or without food without any impairment in efficacy. These studies also showed that ezetimibe was well tolerated, with side effects no different from those seen with placebo. Short-term, early phase 2 studies evaluating the coadministration of ezetimibe and a number of different statins found that coadministration was safe, that ezetimibe did not alter the pharmacokinetics of the statins or vice versa, and that the reductions in low-density lipoprotein cholesterol were complementary, with the degree of cholesterol lowering seen with ezetimibe monotherapy maintained when it was given in combination with a statin. These studies indicate that combination therapy with ezetimibe and a starting dose of a statin produces a reduction in cholesterol levels equivalent to that seen with an 8-fold higher statin dose. Larger, long-term phase 3 trials confirmed the efficacy and safety of the 10-mg dose and also demonstrated that ezetimibe monotherapy is an excellent alternative for patients who cannot tolerate statins. PMID- 11855702 TI - Future outlook: changing perspectives on best practice. AB - The guidelines recently released by the National Cholesterol Education Program call for more aggressive lowering of the level of low-density lipoprotein (LDL) cholesterol and a significant increase in the number of patients eligible for therapy that lowers the level of LDL. Despite the efficacy of statins in lowering the LDL level and in reducing the risk of a coronary event or stroke, other cholesterol-lowering therapies are needed. Some patients are unable to tolerate statins or are not candidates for statin therapy because of liver enzyme abnormalities, age, a preference for nonsystemic therapy, or a modestly elevated LDL level. For those patients, bile acid sequestrants, intestinal bile acid transport inhibitors, acyl coenzyme A:cholesterol acyltransferase inhibitors, and a number of nonselective cholesterol absorption inhibitors are alternative treatments. However, those agents vary in their effectiveness in reducing the level of LDL. Their use often does not reduce the LDL level to the extent desired or is compromised by patients' poor compliance with therapy because of inconvenient dosing or unpleasant side effects. Ezetimibe, the first selective inhibitor of intestinal cholesterol absorption, is a promising alternative to the agents listed above. When ezetimibe is used either as monotherapy or in combination with a statin, once-daily dosing reduces the level of LDL by an average of 18%. PMID- 11855703 TI - Modified guanidines as chiral auxiliaries. AB - Investigations on modified guanidines, prepared by newly developed methods, as potential chiral auxiliaries led to reasonable asymmetric induction not only in catalytic but also in stoichiometric asymmetric syntheses. These guanidine mediated reactions may contribute to the development of green chemistry because of their possible application as re-cyclable (economically favored) and easily functionalizable (widely applicable) auxiliaries. PMID- 11855704 TI - Preparation and characterization of TADDOLs immobilized on hydrophobic controlled pore-glass silica gel and their use in enantioselective heterogeneous catalysis. AB - Highly porous silica gel (controlled-pore glass, CPG, ca. 300 m2 g(-1)) with covalently attached TADDOLs (loading 0.3-0.4 mmol g(-1)) and Me3Si-hydrophobized surface has been prepared: First, mercaptopropyl groups were attached to the silica gel by treatment with (mercaptopropyl)trimethoxysilane; then the SH groups were trityl-protected, and the remaining accessible SiOH groups hydrophobized by silylation (heating with Me3Si-imidazole); after deprotection, the SH groups were used as nucleophiles for benzylation with TADDOLs carrying a 4-bromomethyl-phenyl group in the 2-position of their dioxolane rings; alternatively, the SH groups have been benzylated with the 4-bromomethyl-benzaldehyde acetal of diethyltartrate, and the diarylmethanol moieties of the TADDOLs created on the solid support by addition of excess phenyl, or 1- or 2-naphthyl magnesium bromide. Each step of the immobilizing procedure was carefully monitored and analyzed (Ellman's test, methyl-red test), and resulting materials characterized by electron microscopy, DRIFT spectroscopy (IR), 13C- and 29Si NMR solid-state NMR spectroscopy, and elemental analysis. The immobilized TADDOLs were titanated to give (iPrO)2Ti-, Cl2Ti-, or (TosO)2Ti-TADDOLates which were used for catalyzing the additions of Et2Zn or Bu2Zn to PhCHO and of diphenyl nitrone to 3 crotonoyl-oxazolidinone. The following findings are remarkable: i) The enantioselectivities and conversions of the reactions mediated by the CPG immobilized Ti-TADDOLates match those observed under standard homogeneous conditions. ii) If and when the rates and/or the enantioselectivities of reactions have dropped after several applications of the same catalyst batch, washing with aqueous HCl/acetone and reloading with titanate leads to full restoration of its performance. iii) There is no detectable loss of the hydrophobizing Me3Si groups after nine acidic washes! iv) There is a seasoning of the catalyst material in the Cl2Ti-TADDOLate-mediated [3+2] cycloaddition of diphenylnitrone: Initially it is necessary to use 0.5 equivalents of the immobilized catalyst to match the performance of the homogeneous catalyst; after three runs the reaction rate, enantio- and diastereoselectivity have dropped considerably; acidic washing after each subsequent run completely restores the performance; after a total of seven runs the amount of catalyst can be reduced to 0.4, 0.3, 0.2, and 0.1 equivalents in the following runs, with identical good results! PMID- 11855705 TI - Gamma2-, gamma3-, and gamma(2,3,4)-amino acids, coupling to gamma-hexapeptides: CD spectra, NMR solution and X-ray crystal structures of gamma-peptides. AB - There are numerous possible gamma-amino acids with different degrees of substitution and with various constitutions and configurations. Of these the gamma4- and the like- and unlike-gamma(2,4)-amino acids have been previously used as building blocks in gamma-peptides. The synthesis of gamma2-, gamma3-, and gamma(2,3,4)-peptides is now described. The corresponding amino acids have been prepared by Michael addition of chiral N-acyl-oxazolidinone enolates to nitro olefins, with subsequent reduction of the NO2 to NH2 groups. Such additions to E 2-methyl-nitropropene provide (2R,3R,4R)-2-alkyl-3-methyl-4-amino-pentanoic acid derivatives (9, 10, 11). Stepwise coupling and fragment coupling lead to gamma-di , tri-, and hexapeptides (12-23), which were fully characterized. The crystal structures of one of the gamma-amino acids (2,3-dimethyl-4-amino-pentanoic acid x HCl, 9a), of a gamma(2,3,4)-di- and a gamma(2,3,4)-tetrapeptide (20, 22) are described, and the NMR solution structure in MeOH of a gamma(2,3,4)-hexapeptide (3) has been determined (using TOCSY, COSY, HSOC, HMBC and ROESY measurements and a molecular dynamics simulated-annealing protocol). A linear conformation (sheet like), a novel (M) helix built of nine-membered hydrogen-bonded rings, and (M) 2.6(14) helices have thus been identified. NMR measurements at different temperatures (298-393 K) and H/D-exchange rates obtained for the gamma(2,3,4) hexapeptide are interpreted as evidence for the stability of the 2.6(14) helix (no "melting") and for its non-cooperative folding mechanism. CD Spectra of the gamma-peptides have been measured in MeOH and CH3CN, indicating that only the protected and unprotected gamma(2,3,4)-hexapeptide is present as the 2.6(14) helix in solution. The structures of the gamma2- and gamma3-hexapeptides (1, 2) could not be determined. PMID- 11855706 TI - Supramolecular self-assembling properties of membrane-spanning archaeal tetraether glycolipid analogues. AB - The self-assembling properties of a new series of archaeal tetraether glycolipid analogues 1-6 that are characterized by a bipolar architecture with two similar or different glycosidic and/or phosphate polar heads and a lipid core possessing a cyclopentane unit and/or branched chains were studied by means of differential scanning calorimetry, optical microscopy, X-ray scattering, freeze-fracture electron microscopy and dynamic light scattering. Unsymmetrical phosphate derivatives 1 and 2 spontaneously formed thermostable multilamellar and unilamellar vesicles in which most of the bipolar lipids adopted a trans-membrane conformation, as revealed by freeze-fracture electron microscopy. Supramolecular aggregates of neutral glycolipids 3-6 were found to depend on both the saccharidic polar heads and the chain composition. The presence of one glycosidic residue with rather marked hydrophilic properties, such as the lactosyl moiety, was required to allow the formation of multilamellar vesicles. Surprisingly, the introduction of a cyclopentane unit in the bridging chain was able to induce an apparent two-by-two membrane association: this unusual behaviour might be the result of unsymmetrical interfacial properties of the lipid layer caused by the presence of the cyclopentane unit. PMID- 11855707 TI - Quantitative reaction cascades of ninhydrin in the solid state. AB - Crystalline ninhydrin (1) undergoes waste-free solid-state cascade reactions with dimedone, L-proline, three o-phenylenediamines, o-mercaptoaniline, two ureas, three thioureas, and methyl 3-aminocrotonate. The yields are quantitative and give pure crystalline products without workup just by milling stoichiometric mixtures of the crystalline reagents. The structures of the new and the previously obtained products with lower yields from solutions are established or confirmed by spectroscopic data and density functional calculations at the B3LYP/6-31G* level. The success of 3- and 4-cascade reactions in the crystal without melting is unusual and of unmatched atom economy. They are mechanistically investigated with atomic force microscopy techniques (AFM) on six different faces of 1 when o-phenylenediamine was the reagent (substitution, elimination, cyclization, elimination) and interpreted on the basis of known crystal structure data. Strict correlations to the crystal packings are observed. The characteristic surface features grow to microm heights in some cases at distances of 0.5 mm from the contact edge of the reacting crystals. The waste free and easy syntheses of highly functionalized (C=O; O-H; C=N) heterocycles or of a tetraketone are also of interest for synthetic use. PMID- 11855708 TI - High-resolution EPR spectroscopic investigations of a homologous set of d9 cobalt(0), d9-rhodium(0), and d9-iridium(0) complexes. AB - The 17-electron complexes [M(tropp(ph))2] (M=Co0, Rh0, Ir0) were prepared and isolated (tropp = tropylidene phosphane). A structural analysis of [Co(tropp(ph))2] revealed this complex to be almost tetrahedral, while the heavier homologues have more planar structures. Partially deuterated tropp complexes [D6][M(tropp(ph))2] were synthesised for M = Rh and Ir in order to enhance the resolution in the EPR spectra. This synthesis involves a four-fold intramolecular C-H activation reaction, whereby alkyl groups are transformed into olefins. Dihydrides were observed as intermediates for M = Ir. The electronic and geometric structures of all complexes [M(tropp(ph))2] (M = Co, Rh, Ir) and [D6][M(tropp(ph))2] (M = Rh, Ir) were investigated by continuous wave (CW) and echo-detected EPR in combination with pulse ENDOR and ESEEM techniques. In accord with their planar structures, cis and trans isomers were detected for [M(tropp(ph))2] (M = Rh0, Ir0) for which a dynamic equilibrium was established. The thermodynamic data show that the cis isomer is slightly preferred by deltaH(o) = -4.7 +/- 0.3 kJ mol(-1) (M = Rh) and delta H(o) = -5.1 +/- 0.5 kJ mol(-1); (M = Ir). The entropies for the process trans-[M(tropp(ph))2] <==> cis [M(tropp(ph))2] are also negative [deltaS(o) = -5 +/- 1.5 J mol(-1) (M = Rh); deltaS(o) = -17 +/- 3.7 J mol(-1) (M = Rh)], indicating higher steric congestion in the cis isomers. The cobalt(0) and irdium(0) complexes show rather large g anisotropies, while that of the rhodium(0) complex is small (Co: g(parallel) = 2.320, g(perpendicular) = 2.080; cis-Rh: g(parallel) = 2.030, g(perpendicular) = 2.0135; trans-Rh: g(parallel) = 2.050, g(perpendicular) = 2.030; cis-Ir: g(parallel) = 2.030, g(perpendicular) = 2.060; trans-Ir: g(parallel) = 1.980, g(perpendicular) = 2.150). The g matrices of [M(tropp(ph))2] (M = Co, Rh) are axially symmetric with g(parallel) > g(perpendicular), indicating either a distorted square planar structure (SOMO essentially d(x2 - y2) or a compressed tetrahedron (SOMO essentially d(xy)). Interestingly, for [Ir(tropp(ph))2] the inverse ordering, g(perpendicular) > g(parallel) is found; this cannot be explained by simple ligand field arguments and must await a more sophisticated analysis. The hyperfine interactions of the unpaired electron with the metal nuclei, phosphorus nuclei, protons, deuterons and carbon nuclei were determined. By comparison with atomic constants, the spin densities on these centres were estimated and found to be small. However, the good agreement of the distance between the olefinic protons and the metal centres determined from the dipolar coupling parameter indicates that the unpaired electron is primarily located at the metal centre. PMID- 11855709 TI - Facile preparation of [4.4]metacyclophane- and [5.5]paracyclophane-type macrocycles from arylboronic acids and salicylideneaminoaryl alcohols. AB - Four different salicylideneaminoaryl alcohols have been treated with arylboronic acids in order to prepare air-stable cyclophane-type macrocyclic systems. In two cases, this objective could be realized with the high-yield formation of [4.4]metacyclophane and [5.5]paracyclophane derivatives. The skeleton in these macrocycles is held together by two chiral boron atoms. In the other two cases, monomeric boronates or polymeric material were obtained. The title structures were characterized by spectroscopic techniques and X-ray crystallography. They show transannular C-H...O hydrogen bonding, but no intramolecular pi-pi interactions. A synthetic strategy for the preparation of further boron macrocycles is presented. PMID- 11855710 TI - First use of axially chiral thioamides for the stereocontrol of C-C bond formation. AB - Several N-aryl-substituted thioamides with an axis of chirality along the N C(aryl) bond were prepared in good to excellent yields. NMR spectra revealed preferences for the E rotamer (along the N-C(=S) bond). X-ray crystallographic analysis showed that the planes of the aryl and thioamide groups were almost perpendicular (79 degrees). For the first time, these atropisomeric thioamides were used for an asymmetric Claisen rearrangement. LDA deprotonation led selectively to the enethiolates of Z stereochemistry, and subsequent reaction with a variety of allyl halides furnished S-allyl keteneaminothioacetals. These intermediates were not detected as they rearranged readily to gamma-unsaturated thioamides in good to high yields and diastereoselectivities up to 88:12. Chemical correlation allowed the assignment of the (aR*,2R*) configuration to the major diastereoisomer. A model was proposed to explain the stereochemical course of the thio-Claisen rearrangement. PMID- 11855711 TI - Computational studies on 3-aza-Cope rearrangements: protonation-induced switch of mechanism in the reaction of vinylpropargylamine. AB - The 3-aza-Cope rearrangements of 3-azoniahexa-1,5-diene (1), 3-azoniahex-1-ene-5 yne (3), and 3-azahex-1-ene-5-yne (5) were investigated up to the coupled-cluster level, CCSD(T), by using a valence triple-zeta basis set. Activation barriers and geometrical parameters of the transition states are provided. Conformational studies were performed for all reactants and products of the reactions. Solvent effects were estimated from self-consistent reaction field calculations. In contrast to the other two species, the Cope rearrangement of 5 was found to proceed by a stepwise mechanism. PMID- 11855712 TI - Complexation mechanism of olefin with silver ions dissolved in a polymer matrix and its effect on facilitated olefin transport. AB - Remarkable separation performance of olefin/paraffin mixtures was previously reported by facilitated olefin transport through silver-based polymer electrolyte membranes. The mechanism of facilitated olefin transport in solid membranes of AgCF3SO3 dissolved in poly(N-vinyl pyrrolidone) (PVP) is investigated. In silver polymer electrolyte membranes, only free anions are present up to the 2:1 mole ratio of [C=O]:[Ag], and ion pairs start to form at a ratio of 1:1, followed by higher-order ionic aggregates above a ratio of 1:2. At silver concentrations above 3:1, the propylene permeance increases almost linearly with the total silver concentration, unexpectedly, regardless of the silver ionic constituents. It was also found that all the silver constituents, including ion pairs and higher order ionic aggregates, were completely redissolved into free anions under the propylene environment; this suggests that propylene can be a good ligand for the silver cation. From these experimental findings, a new mechanism for the complexation reaction between propylenes and silver salts in silver-polymer electrolytes was proposed. The new mechanism is consistent with the linearity between the propylene permeance and the total silver concentration regardless of the kind of the silver constituents. Therefore, the facilitated propylene transport through silver-polymer electrolytes may be associated mainly with the silver cation weakly coordinated with both carbonyl oxygen atoms and propylene. PMID- 11855713 TI - 1,1'-Binaphthyl-substituted macrocycles as receptors for saccharide recognition. AB - The preparation of receptors for saccharide recognition in a natural environment has been an unmet goal for a long time. We present herein the synthesis and binding properties of (R,S)-1,1'-binaphthyl-substituted macrocycles as receptors for saccharide recognition in water/acetonitrile (1:1) and in DMSO. Porphyrin and metalloporphyrin macrocycles with two to four 1,1'-binaphthyl substituents and multiple hydroxy groups generate a binding site for saccharides that incorporates hydrogen-bonding hydroxy groups together with the aromatic hydrophobic pocket. The specificity for di- and trisaccharides is governed by the cavity size. The mechanism of binding has been studied by 1H NMR spectroscopy and the role of H bonding and CH-pi interactions has been evaluated; the ability to bind saccharides has been demonstrated by the surface plasmon resonance (SPR) technique. The application of these macrocyclic receptors to sensor development is also presented. PMID- 11855714 TI - Intramolecular allyl transfer reaction from allyl ether to aldehyde groups: experimental and theoretical studies. AB - The intramolecular transfer of the allyl group of functionalized allyl aryl ethers to an aldehyde group in the presence of Ni0 complexes was studied from chemical, electrochemical and theoretical points of view. The chemical reaction involves the addition of Ni0 to the allyl ether followed by stoichiometric allylation. The electrochemical process is catalytic in nickel and involves the reduction of intermediate eta3-allylnickel(II) complexes. PMID- 11855715 TI - Ferrocene-containing carbohydrate dendrimers. AB - Aliphatic amines, incorporating one or three (branched) acylated beta-D glucopyranosyl residues, were coupled with the acid chloride of ferrocenecarboxylic acid and with the diacid chloride of 1,1' ferrocenedicarboxylic acid to afford four dendrimer-type, carbohydrate-coated ferrocene derivatives in good yields (54-92%). Deprotection of the peracylated beta-D-glucopyranosyl residues was achieved quantitatively by using Zemplen conditions, affording four water-soluble ferrocene derivatives. When only one of the two cyclopentadienyl rings of the ferrocene unit is substituted, strong complexes are formed with beta-cyclodextrin in H2O, as demonstrated by liquid secondary ion mass spectrometry (LSIMS), 1H NMR spectroscopy, electrochemical measurements, and circular dichroism spectroscopy. Molecular dynamics calculations showed that the unsubstituted cyclopentadienyl ring is inserted through the cavity of the toroidal host in these complexes. The electrochemical behavior of the protected and deprotected ferrocene-containing dendrimers was investigated in acetonitrile and water, respectively. The diffusion coefficient decreases with increasing molecular weight of the compound. The potential for oxidation of the ferrocene core, the rate constant of heterogeneous electron transfer, and the rate constant for the energy-transfer reaction with the luminescent excited state of the [Ru(bpy)3]2+ complex (bpy = 2,2'-bipyridine) are strongly affected by the number (one or two) of substituents and by the number (one or three) of carbohydrate branches present in the substituents. These effects are assigned to shielding of the ferrocene core by the dendritic branches. Electrochemical evidence for the existence of different conformers for one of the dendrimers in aqueous solution was obtained. PMID- 11855716 TI - Mixed anhydrides: key intermediates in carbamates forming processes of industrial interest. AB - Mixed anhydrides of carbonic acid with phosphonic or carbamic acid, are mimic of relevant biological systems, and behave as key intermediates in trans esterification processes that afford carbamates of industrial interest. They are formed in the phosphonic acids mediated or direct transesterification reaction of organic carbonates with amines to afford carbamates and have been isolated and characterised in the solid state and solution. Their conversion into the products has been demonstrated to occur with high regioselectivity. The application of such intermediates in some synthetic processes is discussed. PMID- 11855717 TI - Alpha-amino acids with metallocenyl side chains. AB - A straightforward method for the synthesis of enantiomerically pure bis(valine)metallocenes is presented. Derivatives of lithium cyclopentadienylvaline 1a, b were obtained by addition of the (R)- or (S) Schollkopf reagents to 6,6-dimethylfulvene as single enantiomers and gave with FeCl2 or [RuCl2(dmso)4] the chiral metallocenes [Fe[C5H4-CMe2 [C4H2N2(OMe)2iPr]]2] (2a, b) and [Ru[C5H4-CMe2-[C4H2N2(OMe)2iPr]]2] (3a, b). Complex 2b was hydrolyzed to the ferrocenylene-bis(valine-methylester) [[Fe[C5H4 CMe2-CH(NH3+)COOMe]2]2+(Cl-)2] (7) without racemization. Complex 7 could be used as ligand and was treated with [[Cp*IrCl2]2] to afford [Fe[C5H4-CMe2 CH(COOMe)(NH2-IrCp*Cl2)]2] (10). The reactions of 1 with CoCl2, [Re(CO)5Br], [[(cod)RhCl2]2] (cod= 1,5-cyclooctadiene) or [Cp*MCl3] (M= Ti, Zr) gave the cyclopentadienyl complexes [[Co[C5H4-CMe2-[C4H2N2(OMe)2iPr]]2]+ I-] (11) and [Re[C5H4-CMe2-[C4H2N2(OMe)2iPr]](CO)3] (13), [(C8H12)Rh[C5H4-CMe2 [C4H2N2(OMe)2(iPr)]]] (14). [[Rh[C5H4-CMe2-[C4H2N2(OMe)2(iPr)]]I]2(mu-I)2] (15), [Cp*Cl2Ti-[C5H4-CMe2-[C4H2N2(OMe)2(iPr)]]] (16), and [Cp*Cl2Zr[C5H4-CMe2 [C4H2N2(OMe)2(iPr)]]] (17), with chiral valine derivatives as substituents on the cyclopentadienyl ring and with excellent diastereoselectivities. Also the Seebach reagent (Boc-BMI) or O'Donnell reagent could be added to 6,6-dimethylfulvene to give the lithium cyclopentadienides Li[C5H4-CMe2-[C3H2(tBu)(N-Boc)(NMe)O]] (18) and Li[C5H4-CMe2-CH(NCPh2)(COOEt)] (21), which formed the ferrocene derivatives [Fe[C5H4-CMe2-[C3H2(tBu)(N-Boc)(NMe)O]]2] (19) and [Fe[C5H4-CMe2 CH(NCPh2)(COOEt)]2] (22). The stable cobaltocinium cation in 11 and the complex 19 could be hydrolyzed to the metallocenes 12 and [Fe(C5H4-CMe2-CH(NH3+)(COO-)]2] (20) with two valines in the 1,1'-position. The structures of 2a, b, 11, 15, and 16 were determined by X-ray diffraction and confirm the diastereomeric purity of the compounds. PMID- 11855718 TI - Superweak complexes of tetrahedral P4 molecules with the silver cation of weakly coordinating anions. AB - The silver aluminates AgAl[OC(CF3)2(R)]4 (R = H, CH3, CF3) react with solutions of white phosphorus P4 to give complexes that bind one or two almost undistorted tetrahedral P4 molecules in an fashion: [Ag(P4)2]+[Al(OC(CF3)3)4]+ (1) containing the first homoleptic metal-phosphorus cation, the molecular species (P4)AgAl[OC(CH3)(CF3)2]4 (2), and the dimeric Ag(mu,eta2-P4)Ag bridged [(P4)AgAl[OC(H)(CF3)2]4]2 (3). Compounds 1-3 were characterized by variable temperature (VT) 31P NMR spectroscopy (1 also by VT 32P MAS-NMR spectroscopy), Raman spectroscopy, and single-crystal X-ray crystallography. Other Ag:P4 ratios did not lead to new species, and this observation was rationalized on thermodynamic grounds. The Ag(P4)2+ ion has an almost planar coordination environment around the Ag+ ion due to d(x2 - y2)(Ag) --> sigma*(P-P) backbonding. Calculations (HF-DFT) on six Ag(P4)2+ isomers 4a-f showed that the planar eta2 form 4a is only slightly favored by 5.2 kJ mol(-1) over the tetrahedral eta2 species 4b; eta1-P4 and eta3-P4 complexes are less favorable (27-76 kJ mol(-1)). The bonding of the P4 moiety in [RhCl(eta2-P4)(PPh3)2], the only compound in which an eta2 bonding mode of a tetrahedral P4 molecule has been claimed, must be regarded as a tetraphosphabicyclobutane, and not as a tetrahedro-P4 complex, on the basis of the published NMR and vibrational spectra, the calculated geometry of [RhCl(P4)(PH3)2] (10), the highly endothermic (385 kJ mol(-1)) calculated dissociation enthalpy of 10 into P4 and RhCl(PH3)2 (11), as well as atoms in molecules (AIM) and natural bond orbital (NBO) population analyses of 10 and the Ag(P4)2+ ion. Therefore, 1-3 are the first examples of species containing eta2 coordinated tetrahedral P4 molecules. PMID- 11855719 TI - Alpha-LNA (locked nucleic acid with alpha-D-configuration): synthesis and selective parallel recognition of RNA. AB - Alpha-LNA is presented as a stereoisomer of LNA (locked nucleic acid) with alpha D-configuration. Three different approaches towards the thymine alpha-LNA monomer as well as the 5-methylcytosine alpha-LNA monomer are presented. Different alpha LNA sequences have been synthesised and their hybridisation with complementary DNA and RNA has been evaluated by means of thermal stability experiments and circular dichroism spectroscopy. In a mixed pyrimidine sequence, alpha-LNA displays unprecedented parallel-stranded and selective RNA binding. Furthermore, a remarkable selectivity for hybridisation with RNA over DNA is indicated. PMID- 11855720 TI - Self-assembly of gold(I) rings and reversible formation of organometallic [2]catenanes. AB - The reaction of the digold(I) diacetylide [(AuCCCH2OC6H4)2CMe2] with diphosphane ligands can lead to formation of either macrocyclic ring complexes or [2]catenanes by self-assembly. This gives an easy route to rare organometallic [2]catenanes, and the effect of the diphosphane ligand on the selectivity of self assembly is studied. With diphosphane ligands Ph2P(CH2)xPPh2, the simple ring complex [Au2[(CCCH2OC6H4)2CMe2](Ph2P(CH2)xPPh2)] is formed selectively when x = 2, but the [2]catenanes [Au2[(CCCH2OC6H4)2CMe2](Ph2P(CH2)xPPh2)]2 are formed when x = 4 or 5. When x = 3, a mixture of the simple ring and [2]catenane is formed, along with the "double-ring" complex, [Au4[(CCCH2OC6H4)2CMe2]2(Ph2P(CH2)3PPh2)2] and a "hexamer" Au2[(CCCH2OC6H4)2CMe2](Ph2P(CH2)3PPh2)]6] whose structure is not determined. A study of the equilibria between these complexes by solution NMR techniques gives insight into the energetics and mechanism of [2]catenane formation. When the oligomer [(AuCCCH2OC6H4)2CMe2] was treated with a mixture of two diphosphane ligands, or when two [2]catenane complexes [[Au2[(CCCH2OC6H4)2CMe2](diphosphane)]2] were allowed to equilibrate, only the symmetrical [2]catenanes were formed. The diphosphanes Ph2PCCPPh2, trans [Ph2PCH=CHPPh2] and (Ph2PC5H4)2Fe give the corresponding ring complexes [Au2[(CCCH2OC6H4)2CMe2](diphosphane)], and the chiral, unsymmetrical diacetylide [Au2[(CCCH2OC6H4C(Me)(CH2CMe2)C6H3OCH2CC)] gives macrocyclic ring complexes with all diphosphane ligands Ph2P(CH2)xPPh2 (x = 2-5). PMID- 11855721 TI - Alternating right- and left-handed helical loops in a self-assembled polymer: direct observation of ring-opening polymerization of a macrocyclic gold complex. AB - Reaction of the ligand NN'-bis(pyridin-3-yl)-1,3-benzenedicarboxamide (1) with the diphosphanedigold(I) complex [(mu-PP)(AuO2CCF3)2] occurred by displacement of the trifluoroacetate ligands by the pyridyl groups of 1, and crystallization gave the macrocyclic complex [Au2(mu-PP)(mu-1)](CF3CO2)2 (2b), when PP = trans [Ph2PCH=CHPPh2] but the polymer [(Au2(mu-PP)(mu-1)]x](CF3CO2)2x (3), when PP= Ph2PCH2CH2PPh2. The polymer 3 contains a series of helical turns connected by linear chain sections. and the helices have alternating right- and left-handed conformations. The polymer 3 dissolves to give an equilibrium mixture of the macrocyclic ring complex [Au2(mu-PP)(mu-1)](CF3CO2)2 (2a), and ring-opened oligomers, thus giving insight into the mechanism of the ring-opening polymerization reaction. PMID- 11855722 TI - The subcellular localization of an unusual rice calmodulin isoform, OsCaM61, depends on its prenylation status. AB - Calmodulin (CaM) is a small Ca2+-binding protein highly conserved in eukaryotes. We have reported previously a novel rice CaM-like protein (OsCaM61) which contains an N-terminal CaM domain and a C-terminal extension with a potential prenylation site. Here we report in vitro activity assays confirm OsCaM61 as a functional CaM. Using the green fluorescent protein (GFP) as a visual marker, we further studied the subcellular localization of OsCaM61 in stably transformed tobacco cells. The GFP-OsCaM61 fusion protein was membrane-associated whereas OsCaM61-GFP was mainly detected in the nucleoplasm. GFP-OsCaM61 was transported into the nucleoplasm upon a block in isoprenoid biosynthesis by mevinolin treatment of cells. These results indicate that the prenylated OsCaM61 molecules are mainly membrane-associated whereas its unprenylated counterparts are transported into the nucleoplasm. Thus, OsCaM61 may play functions in co ordinating Ca2+ signaling with isoprenoid metabolism. PMID- 11855723 TI - NADP-malate dehydrogenase from Chlamydomonas: prediction of new structural determinants for redox regulation by homology modelling. AB - The function of a gene closely linked to nitrate assimilation loci from Chlamydomonas reinhardtii has been investigated. Gene expression analysis shows that its mRNA accumulation is modulated by light, carbon source and adaptation to light/dark cyclic conditions of growth. A full-length cDNA was isolated for the light-regulated transcript, and sequence characterization indicates that it encodes the NADP-malate dehydrogenase from C. reinhardtii (NADP-MDH;Cr). The primary structure of NADP-MDH;Cr is closely related to plant, mossfern and algal NADP-malate dehydrogenases, and shares structural determinants for chloroplast targeting, cofactor binding and catalysis. Sequence conservation extends to the carboxy end of the protein, where plant and mossfern enzymes have two cysteines and an acidic C-terminus with a critical role for regulation of NADP-MDH activity by the thioredoxin/ferredoxin system. Accordingly, incubation with DTT activates NADP-MDH enzyme in cell-free extracts from C. reinhardtii. Like NADP-malate dehydrogenases from two other green algae, the N-terminal extension of NADP MDH;Cr lacks two thiol residues whose reduction constitutes the rate-limiting step in the activation reaction of plant enzymes. Homology-based 3D modelling of NADP-MDH;Cr, the first structure predicted for NADP-malate dehydrogenase from a lower eukaryote, evidences close positioning of two new cysteines in an accessible region of the protein surface. These results suggest that the algal enzyme has a different arrangement of regulatory disulfide bridges, which might involve the existence of new mechanisms that control functioning of the malate valve, the main system to export reducing power from the chloroplast of plant cells. PMID- 11855724 TI - Wound-response regulation of the sweet potato sporamin gene promoter region. AB - Sporamin, a tuberous storage protein of sweet potato, was systemically expressed in leaves and stems by wound stimulation. In an effort to demonstrate the regulatory mechanism of wound response on the sporamin gene, a 1.25 kb sporamin promoter was isolated for studying the wound-induced signal transduction. Two wound response-like elements, a G box-like element and a GCC core-like sequence were found in this promoter. A construct containing the sporamin promoter fused to a beta-glucuronidase (GUS) gene was transferred into tobacco plants by Agrobacterium-mediated transformation. The wound-induced high level of GUS activity was observed in stems and leaves of transgenic tobacco, but not in roots. This expression pattern was similar to that of the sporamin gene in sweet potatoes. Exogenous application of methyl jasmonate (MeJA) activated the sporamin promoter in leaves and stems of sweet potato and transgenic tobacco plants. A competitive inhibitor of ethylene (2,5-norbornadiene; NBD) down-regulated the effect of MeJA on sporamin gene expression. In contrast, salicylic acid (SA), an inhibitor of the octadecanoid pathway, strongly suppressed the sporamin promoter function that was stimulated by wound and MeJA treatments. In conclusion, wound response expression of the sporamin gene in aerial parts of plants is regulated by the octadecanoid signal pathway. PMID- 11855725 TI - Conservation in divergent solanaceous species of the unique gene structure and enzyme activity of a gametophytically-expressed flavonol 3-O galactosyltransferase. AB - Flavonol 3-O-galactosyltransferase (F3GalTase) is a pollen-specific enzyme which glycosylates the flavonols required for germination in petunia. The highly restricted tissue-specific expression and substrate usage make F3GalTase unique among all other flavonoid glycosyltransferases (GTs) described to date, including the well characterized Bronze 1 (Bz1) gene of maize. RFLP mapping, DNA gel blot, and sequence analyses showed that the single copy F3galtase gene has a different genomic organization than Bz1. Within the promoter of F3galtase are potential regulatory motifs which may confer pollen-specific gene expression and activation by Myb and bHLH transcription factors. However, we provide evidence that F3galtase is not regulated by An4, which encodes a Myb factor known to regulate anthocyanin accumulation in petunia anthers. An unexpected feature of the F3galtase promoter was the presence of large blocks of chloroplast and mitochondrial DNA. Gel blot analyses of genomic DNA from the progenitors of Petunia hybrida, as well as from Nicotiana tabacum, indicated that migration of organellar DNA into the F3galtase gene was an ancient event that occurred prior to speciation of the Solanaceae. Together with enzyme assays and HPLC analyses of pollen extracts from tobacco, tomato, and potato, these results confirmed that the unique F3galtase gene structure, enzyme activity, and pollen-specific flavonol glucosylgalactosides are conserved throughout the Solanaceae. PMID- 11855726 TI - Induction of pepper cDNA encoding a lipid transfer protein during the resistance response to tobacco mosaic virus. AB - Pepper (Capsicum annuum) plants exhibit hypersensitive response (HR) against infection by many tobamoviruses. A clone encoding a putative nonspecific lipid transfer protein (CaLTP1) was isolated by differential screening of a cDNA library from resistant pepper leaves when inoculated with tobacco mosaic virus (TMV) pathotype P0. The predicted amino acid sequence of CaLTP1 is highly similar to that of the other plant LTPs. Southern blot analysis showed that a small gene family of LTP-related sequences was present in the pepper genome. Transcripts homologous to CaLTP1 accumulated abundantly in old leaves and flowers. CaLTP1 expression was induced in the incompatible interaction with TMV-P0 but was not induced in the compatible interaction with TMV-P1.2. In correlation with the temporal progression of HR in the inoculated leaves, CaLTP1 transcripts started to accumulate at 24 h after TMV-P0 inoculation, reaching a maximal level at 48 h. A strain of Xanthomonas campestris pv. vesicatoria (Xcv) that carries the bacterial avirulence gene, avrBs2, was infiltrated into leaves of a pepper cultivar containing the Bs2 resistance gene. A marked induction of CaLTP1 expression was observed in Xcv-infiltrated leaves. Effects of exogenously applied abiotic elicitors on CaLTP1 expression were also examined. Salicylic acid caused a rapid accumulation of CaLTP1 transcripts in pepper leaves and ethephon treatment also induced the expression of the CaLTP1 gene. Transient expression in the detached pepper leaves by biolistic gene bombardment indicated that CaLTP1 is localized mostly at the plant cell surface, possibly in the cell wall. These results suggest possible role(s) for LTPs in plant defense against pathogens including viruses. PMID- 11855727 TI - Cloning and characterization of a cDNA encoding a cobalamin-independent methionine synthase from potato (Solanum tuberosum L.). AB - A potato cDNA clone, StMS1, that encodes a methionine synthase was isolated. This protein was identified on the basis of both structural and functional evidence. The predicted sequence of the protein encoded by StMS1 shows a high degree of similarity to methionine synthases from other organisms and the expression of StMS1 in bacterial mutant strains restored the mutant's ability to synthesize methionine. Genomic organization and expression analyses suggest that StMS1 is a low-copy gene and is differentially expressed in potato organs. StMS1 expression was found in all tissues, but at elevated levels in flowers, basal levels in sink and source leaves, roots and stolons, and low levels in stems and tubers. RNA expression data were confirmed by western blot analysis except that the protein content in leaves was less than expected from the RNA data. Western blot analysis of subcellular fractions revealed that the protein is located in the cytosol. However, the changing pattern of gene expression during the day/night period implied a light-dependent control of MS transcription normally seen for enzymes localized in plastids. The expression of MS was shown to be light-inducible with its highest expression at midday. These RNA data were not confirmed at the protein level since protein content levels remained constant over the whole day. Feeding experiments of detached leaves revealed that sucrose or sucrose-derived products are responsible for StMS1 induction. This induction can be blocked by treatment with DCMU during the light period. Western analysis revealed that the amount of StMS1 is not affected by either treatment. This experiment confirmed the presence of a day/night rhythm. Methionine synthase expression is regulated by photoassimilates but this seems not to detectably alter protein levels. PMID- 11855728 TI - Infection of Arabidopsis with a necrotrophic pathogen, Botrytis cinerea, elicits various defense responses but does not induce systemic acquired resistance (SAR). AB - Botrytis cinerea is a non-specific necrotrophic pathogen that attacks more than 200 plant species. In contrast to biotrophs, the necrotrophs obtain their nutrients by first killing the host cells. Many studies have shown that infection of plants by necrosis-causing pathogens induces a systemic acquired resistance (SAR), which provides protection against successive infections by a range of pathogenic organisms. We analyzed the role of SAR in B. cinerea infection of Arabidopsis. We show that although B. cinerea induced necrotic lesions and camalexin biosynthesis, it did not induce SAR-mediated protection against virulent strains of Pseudomonas syringae, or against subsequent B. cinerea infections. Induction of SAR with avirulent P. syringae or by chemical treatment with salicylic acid (SA) or benzothiadiazole also failed to inhibit B. cinerea growth, although removal of basal SA accumulation by expression of a bacterial salicylate hydroxylase (NahG) gene or by infiltration of 2-aminoindan-2 phosphonic acid, an inhibitor of phenylpropanoid pathway, increased B. cinerea disease symptoms. In addition, we show that B. cinerea induced expression of genes associated with SAR, general stress and ethylene/jasmonate-mediated defense pathways. Thus, B. cinerea does not induce SAR nor is it affected by SAR, making it a rare example of a necrogenic pathogen that does not cause SAR. PMID- 11855729 TI - cDNA cloning and expression of carotenogenic genes during flower development in Gentiana lutea. AB - All cDNAs involved in carotenoid biosynthesis leading to lycopene in yellow petals of Gentiana lutea have been cloned from a cDNA library. They encode a geranylgeranyl pyrophosphate synthase, a phytoene synthase, a phytoene desaturase and a zeta-carotene desaturase. The indicated function of all cDNAs was established by heterologous complementation in Escherichia coli. The amino acid sequences deduced from the cDNAs were between 47.5% and 78.9% identical to those reported for the corresponding enzymes from other higher plants. Southern analysis suggested that the genes for each enzyme probably represent a small multi-gene family. Tissue-specific expression of the genes and expression during flower development was investigated. The expression of the phytoene synthase gene, psy, was enhanced in flowers but transcripts were not detected in stems and leaves by northern blotting. Transcripts of the genes for geranylgeranyl pyrophosphate (ggpps), phytoene desaturase (pds) and zeta-carotene desaturase (zds) were detected in flowers and leaves but not in stems. Analysis of the expression of psy and zds in petals revealed that levels of the transcripts were lowest in young buds and highest in fully open flowers, in parallel with the formation of carotenoids. Obviously, the transcription of these genes control the accumulation of carotenoids during flower development in G. lutea. For pds only a very slight increase of mRNA was found whereas the transcripts of ggpps decreased during flower development. PMID- 11855730 TI - Identification of Mutator insertional mutants of starch-branching enzyme 1 (sbe1) in Zea mays L. AB - Starch-branching enzymes (SBE) alter starch structure by breaking an alpha-1,4 linkage and attaching the reducing end of the new chain to a glucan chain by an alpha 1,6 bond. In maize, three isoforms of SBE have been identified. In order to examine the function of the SBEI isoform, a reverse-genetics PCR-based screen was used to identify a mutant line segregating for a Mutator transposon within Sbe1. Compared to wild-type controls, Sbe1 transcripts accumulate at extremely low levels in leaves of the homozygous mutant. Antibodies failed to detect SBEI in leaf tissue of mutants or wild-type controls. In contrast, the level of SBEI in endosperm is undetectable in homozygous mutants while easily detected in wild type controls. Starches extracted from mutant leaves and endosperm have structures indistinguishable from starches of wild-type controls as determined by size-exclusion chromatography (SEC) of intact starch and high-performance SEC of debranched starch. To investigate the possibility of compensation for the lack of SBEI by expression of the homologous sequence reported by Kim etal. (1998), agenomic fragment (Sbe1b) of this sequence was cloned. Northern hybridizations of mutant leaf, root, tassel, endosperm and embryo tissues with non-specific Sbelb probes failed to reveal expression of the homologous sequence. These results suggest that the homologous sequence is not compensating for a lack of SBEI in sbe1::Mu mutants. Further study of this sbel mutation in the presence of other genetic mutations may help to understand the role of SBEI in determining starch structure in leaves and endosperm. PMID- 11855731 TI - Molecular cloning, expression and characterization of tropinone reductase II, an enzyme of the SDR family in Solanum tuberosum (L.). AB - Calystegines are nortropane alkaloids that are found in Solanaceae containing the classical tropane alkaloids hyoscyamine and scopolamine, and in other Solanaceae such as potato, Solanum tuberosum (L.). Calystegines are assumed to derive from the classical tropane alkaloid pathway. We isolated a cDNA from S. tuberosum with high homology to the pseudotropine-forming tropinone reductase (TRII), which presents as the first putative metabolite specific to calystegines. The equivalent amino acid sequence shows typical motifs of a short-chain dehydrogenase (SDR). The recombinant TRII protein expressed in Escherichia coli catalyzes pseudotropine formation from tropinone with a Km value, a pH optimum, substrate and co-substrate preferences similar to those reported for the TRII enzymes from other Solanaceae species. The gene is expressed in roots, tubers and aerial parts of potato. The distribution of the TRII transcript in comparison with the calystegine concentrations in the tissues suggests transport of calystegines or their precursors between potato organs. PMID- 11855732 TI - Inhibitory effect of myb7 uORF on downstream gene expression in homologous (rice) and heterologous (tobacco) systems. AB - The rice myb7 mRNA contains in its long leader an upstream open reading frame (uORF) putatively coding for a 40 amino acid peptide. uORFs have been found in the leader of mRNAs encoding transcriptional factors or other proteins involved in cellular growth and development. They are thought to translationally regulate the expression of downstream ORFs. Here, we showed the ability of the myb7 uORF to inhibit the expression of downstream reporter genes both in homologous (rice) and heterologous (tobacco) systems. This effect seems partially related to its translation, as indicated by the comparison with the mutagenized uORF. In both systems most of the inhibitory effect was due to the presence of the intercistronic region, in disagreement with the Kozak model. Moreover, replacing the uORF or the intercistronic region with a different one, we demonstrated that the inhibitory effect strictly depends on their co-presence. Finally, in vitro assays showed that the myb7 uORF is translated and inhibits the downstream ORF translation. PMID- 11855734 TI - The use of calcium blockers to study biochemical behaviour of Saccharomyces cerevisiae cells. AB - Calcium channel blockers, currently used in clinical medicine, from the dihydropyridine class (nifedipine and nimodipine) were used to study biochemical behaviour of Saccharomyces cerevisiae cells. At a concentration of 200 microM, both drugs partially blocked calcium uptake from an extracellular medium. On the other hand, nifedipine (300 microM) induced a collapse of the membrane potential (delta psi) of these cells, while nimodipine, at the same concentration, produced a similar but less intense effect. Probably, the small difference in the chemical structure between these drugs may explain the same effect with different intensity. With these experiments was possible to observe that these microorganisms have type 'L' calcium channels. PMID- 11855733 TI - Three classes of proteinase inhibitor gene have distinct but overlapping patterns of expression in Pisum sativum plants. AB - Genes representative of three gene classes encoding proteinase inhibitor proteins, with distinct spatial expression patterns, were isolated and characterized from Pisum. Under standard plant growth conditions, one class is expressed exclusively in seeds, whereas the other two make minor contributions to seed inhibitor proteins but are also expressed in other organs, predominantly in root endodermal and floral reproductive tissues. Two of the gene classes contain few genes and are genetically linked at the Tri locus, whereas the third class displays complex hybridization patterns to genomic DNA and maps to diverse genetic loci. Expression analysis of this last class suggests that only a small number of these genes are expressed. The quantitative effect of the Tri locus on root and floral inhibitor gene expression was examined in near-isogenic lines of pea. The proteins encoded by the three classes are all members of the same family (Bowman-Birk) of enzyme inhibitors but are distinct in terms of overall sequence, active site sequences and inhibitor function. PMID- 11855735 TI - Inhibition of growth and cell cycle arrest of ARCaP human prostate cancer cells by ectopic expression of ER-alpha. AB - The estrogen receptor-alpha (ER-alpha) is a ligand-dependent transcription factor that regulates the growth, differentiation, and development of hormone-responsive target organs. While ER-alpha has been reported to play critical role in the pathogenesis and prognosis of breast and prostate cancers, its possible functional role in regulating prostate cancer cell growth in a ligand-dependent or -independent manner is poorly understood. We addressed this question by stably transfecting wild type (wt) ER-alpha cDNA into an invasive estrogen receptor negative human prostate cancer cell line ARCaP. We isolated several clonal lines of transfected cells expressing varying levels of ER-alpha. The ectopic expression of wt ER-a markedly inhibited the growth of ARCaP cells in vitro in an ER-a dose-dependent but ligand-independent manner. Flow cytometric analysis of the wt ER-alpha-transfected ARCaP cells revealed that wt ER-alpha expression arrested cell growth in G1 phase. Our results suggest that ER-alpha may regulate prostate cell growth and participate in the pathogenesis of prostate cancer. ER alpha may be delivered and expressed ectopically to target prostate cancer progression. PMID- 11855736 TI - Antifungal activities of origanum oil against Candida albicans. AB - The antimicrobial properties of volatile aromatic oils from medicinal as well as other edible plants has been recognized since antiquity. Origanum oil, which is used as a food flavoring agent, possesses a broad spectrum of in vitro antimicrobial activities attributed to the high content of phenolic derivatives such as carvacrol and thymol. In the present study, antifungal properties of origanum oil were examined both in vitro and in vivo. Using Candida albicans in broth cultures and a micro dilution method, comparative efficacy of origanum oil, carvacrol, nystatin and amphotericin B were examined in vitro. Origanum oil at 0.25 mg/ml was found to completely inhibit the growth of C. albicans in culture. Growth inhibitions of 75% and >50% were observed at 0.125 mg/ml and 0.0625 mg/ml level, respectively. In addition, both the germination and the mycelial growth of C. albicans were found to be inhibited by origanum oil and carvacrol in a dose dependent manner. Furthermore, the therapeutic efficacy of origanum oil was examined in an experimental murine systemic candidiasis model. Groups of mice (n = 6) infected with C. albicans (5 x LD50) were fed varying amounts of origanum oil in a final vol. of 0.1 ml of olive oil (vehicle). The daily administration of 8.6 mg of origanum oil in 100 microl of olive oil/kg body weight for 30 days resulted in 80% survivability, with no renal burden of C. albicans as opposed to the group of mice fed olive oil alone, who died within 10 days. Similar results were obtained with carvacrol. However, mice fed origanum oil exhibited cosmetically better clinical appearance compared to those cured with carvacrol. The results from our study encourage examination of the efficacy of origanum oil in other forms of systemic and superficial fungal infections and exploration of its broad spectrum effect against other pathogenic manifestations including malignancy. PMID- 11855738 TI - Corticosteroid-binding globulin synthesis and distribution in rat white adipose tissue. AB - Corticosterone binding (CB) capacity was determined in visceral and subcutaneous white adipose tissue (WAT), as well as in plasma of lean Zucker rats. Perfusion of rats with saline eliminated most liver and kidney corticosterone binding but did not affect CB in WAT. The cytosol extracts of isolated cells, however, did not bind corticosterone in detectable amounts. By means of a RT-PCR procedure it was found that corticosterone-binding globulin (CBG) was expressed in WAT. By immunohistochemical detection in WAT sections, CBG was seen in a thin layer surrounding the cells near the plasma membrane. These data suggest that the CBG layer surrounding the cells may act as a protective barrier limiting the access of glucocorticoids to adipocytes. PMID- 11855737 TI - Bilirubin binding to normal and modified human erythrocyte membranes: effect of phospholipases, neuraminidase, trypsin and CaCl2. AB - Binding of bilirubin to human erythrocyte membranes was studied after various enzymatic treatments as well as calcium loading. Whereas phospholipase D treatment of erythrocyte membranes resulted in 23% increase in bilirubin binding, phospholipase C-treated membranes showed remarkable enhancement in bilirubin binding. Polar head groups in general and negatively charged phosphate moieties, in particular, of phospholipids of the membrane appear to inhibit a large amount of bilirubin from binding to the membranes. Neuraminidase treatment of the membranes also led to a slight increase in bilirubin binding as compared to untreated membranes. Membrane proteins and carbohydrates seem to play significant regulatory role in bilirubin binding. However, no direct correlation was found between the increase in bilirubin binding and the amount of carbohydrate released upon tryptic digestion of membranes. Increase in bilirubin binding to trypsin treated membranes can be ascribed to the increase in free bilirubin concentration in the incubation mixture as a result of tryptic hydrolysis of albumin by the membrane-bound tryptic activity. Calcium-loaded erythrocyte membranes also showed remarkable increase in bilirubin binding as a result of negative charge shielding and calcium-induced hydrophobic aggregation. Taken together, these results suggest the inhibitory role of polar head groups of phospholipids (phosphate in particular), carbohydrate and sialic acid in the binding of bilirubin to erythrocyte membranes. PMID- 11855739 TI - Diurnal rhythm of cholesterol biosynthesis in experimental chronic renal failure. AB - Changes in lipid metabolism are an important risk factor for vascular complications during chronic renal failure (CRF). In experimental CRF hypercholesterolemia has been found to be the main lipid disorder. It is probably due to enhanced cholesterologenesis. Mechanisms of these changes remain poorly understood. It is well known that activity of cholesterologenesis undergoes a significant diurnal rhythm. However, there was no evidence that this rhythm is still present in the course of experimental CRF. Results of our studies indicate that in contrast to puromycin induced nephrotic syndrome, diurnal rhythm of cholesterologenesis in CRF rats is preserved both in liver and in the intestine tissue. Significant higher incorporation of tritiated water into cholesterol fraction was found in vivo both in liver as well as in intestine of CRF rats, as compared to control animals. Increased (with comparison to the controls) incorporation of 14C-acetate, and 3H-mevalonate into CRF rat liver sterols indicate that mechanism of enhanced cholesterologenesis is more complex than simply due to the elevated level of mevalonate (potential substrate for cholesterologenesis) which has been reported in plasma of CRF animals. PMID- 11855741 TI - Contractile function of rat myocardium is less susceptible to hypoxia/reoxygenation after acute infarction. AB - In this study we tested the hypothesis that induction of heat shock proteins (HSPs) and antioxidant enzymes is a compensatory mechanism, which preserves the contractility of the surviving myocardium after acute myocardial infarction. For this purpose, mechanical function of isolated rat papillary muscles was tested 15 h after experimental myocardial infarction and sham operation, respectively. Contractility of the preparations was compared to the expression of HSP25, HSP72, and glutathione peroxidase activity (GSH-Px) at normoxia and during hypoxia/reoxygenation. At normoxic conditions, rates of isometric contraction and, in particular, of relaxation were significantly higher after acute myocardial infarction than after sham operation. Improved relaxation rates were reflected in 2- to 3-fold higher heat shock protein levels in papillary muscles from rats with myocardial infarction compared to sham operated animals. During hypoxia/reoxygenation, the rates of contraction and relaxation were better preserved after myocardial infarction than after sham surgery. Recovery of relaxation rates during reoxygenation was associated with increased HSP25 levels and enhanced GSH-Px activity after myocardial infarction. In conclusion, heat shock proteins exert a beneficial effect on cardiac muscle relaxation after acute myocardial infarction. Enhanced heat shock protein expression and GSH-Px activity may protect the contractile function of the surviving myocardium against the damaging influence of hypoxia/reoxygenation during the early post-infarct period. PMID- 11855740 TI - Inhibitory effect of menaquinone-7 (vitamin K2) on osteoclast-like cell formation and osteoclastic bone resorption in rat bone tissues in vitro. AB - The effect of menaquinone-7 (MK-7; vitamin K2) on osteoclast-like cell formation and osteoclastic bone resorption in rat femoral tissues in vitro was investigated. The bone marrow cells were cultured for 7 days in a a-minimal essential medium (alpha-MEM) containing a well-known bone resorbing agent [parathyroid hormone (1-34) (PTH) or prostaglandin E2 (PGE2)] with an effective concentration. Osteoclast-like cells were estimated by staining for tartrate resistant acid phosphatase (TRACP), a marker enzyme of osteoclasts. The presence of PTH (10(-8) M) or PGE2 (10(-6) M) induced a remarkable increase in osteoclast like multinucleated cells. These increases were significantly inhibited by MK-7 (10(-8) - 10(-5) M). MK-7 (10(-7) and 10(-6) M) significantly inhibited phorbol 12-myristate 13-acetate-induced osteoclast-like cell formation, whereas MK-7 did not inhibit dibutyryl cyclic adenosine monophosphate (DcAMP) (10(-5) M)-induced osteoclast-like cell formation. These results suggest that the inhibitory action of MK-7 is partly involved in protein kinase C signaling. The bone cells isolated from rat femoral tissues were cultured for 48 h in an alpha-MEM containing either vehicle or MK-7 (10(-8) - 10(-5) M). The presence of MK-7 (10(-6) and 10(-5) M) caused a significant decrease in the number of mature osteoclasts. Such a decrease was also seen in the presence of calcitonin (10(-10) - 10(-8) M), DcAMP (10(-6) and 10(-5) M), or calcium chloride (10(-4) and 10(-3) M). The effect of MK-7 (10(-6) M) in decreasing the number of osteoclasts was not further enhanced in the presence of calcitonin (10(-8) M), DcAMP (10(-5) M), or calcium chloride (10(-3) M), and was completely abolished by the presence of dibucaine (10(-6) M) or staurosporine (10(-7) M), which are inhibitors of Ca2+-dependent protein kinases. These results suggested that MK-7 has a suppressive effect on osteoclasts. Moreover, the femoral-metaphyseal tissues obtained from rats were cultured for 48 h in Dulbecco's modified Eagle's medium containing either vehicle, PTH (10(-7) M), orPGE2 (10(-5) M) in the absence or presence of MK-7 (10(-7) - 10(-5) M). The presence of PTH or PGE2 induced a significant decrease in bone calcium content. These decreases were significantly blocked by MK-7 (10( 7) - 10(-5) M). This study demonstrates that MK-7 has an inhibitory effect on osteoclastic bone resorption in vitro. PMID- 11855742 TI - Identification and characterization of muscarinic acetylcholine receptor subtypes expressed in human skin melanocytes. AB - The present study was designed to identify and characterize muscarinic acetylcholine receptors in normal human melanocytes. We used subtype-specific oligonucleotide primers to localize the five genetically defined mAChR mRNAs (ml through m5) by reverse transcription-polymerase chain reaction. These experiments showed that all five mAChR subtype mRNAs are expressed in melanocytes. The PCR products were verified by restriction analysis and Southern blotting. Receptors were visualized in cultures of normal human melanocytes and specimens of normal human skin by subtype-specific rabbit anti-receptor polyclonal antibodies. Radioligand binding assays with the lipophilic drug [3H]quinuclidinyl benzilate demonstrated approximately 9,000 high affinity binding sites/cell. Micromolar concentrations of muscarine or carbachol transiently increased intracellular Ca2+, which could be attenuated by atropine, demonstrating coupling of the receptors to mobilization of intracellular free Ca2+. Lower concentrations of muscarine induced spontaneous repetitive spike-like increases of intracellular Ca2+ which is characteristic for the activation of muscarinic receptors. These results indicate that normal human skin melanocytes express the ml, m2, m3, m4, and m5 subtypes of classic muscarinic acetylcholine receptors on their cell membrane and that these receptors regulate the concentration of intracellular free Ca2+, which may play an important physiologic role in melanocyte behavior and skin pigmentation. PMID- 11855743 TI - Pyridoxal isonicotinoyl hydrazone (PIH) prevents copper-mediated in vitro free radical formation. AB - Pyridoxal isonicotinoyl hydrazone (PIH) is an iron chelator with antioxidant activity, low toxicity and is useful in the experimental treatment of iron overload diseases. Previous studies on x-ray diffraction have revealed that PIH also forms a complex with Cu(II). Since the main drug of choice for the treatment of Wilson's disease, d-penicillamine, causes a series of side effects, there is an urgent need for the development of alternative copper chelating agents for clinical use. These chelators must also have antioxidant activity because oxidative stress is associated with brain and liver copper-overload. In this work we tested the ability of PIH to prevent in vitro free radical formation mediated by Cu(II), ascorbate and dissolved O2. Degradation of 2-deoxyribose mediated by 10 microM Cu(II) and 3 mM ascorbate was fully inhibited by 10 microM PIH (I50 = 6 microM) or 20 microM d-penicillamine (I50 = 10 microM). The antioxidant efficiency of PIH remained unchanged with increasing concentrations (from 1 to 15 mM) of the hydroxyl radical detector molecule, 2-deoxyribose, indicating that PIH does not act as a hydroxyl scavenger. On the other hand, the efficiency of PIH (against copper-mediated 2-deoxyribose degradation and ascorbate oxidation) was inversely proportional to the Cu(II) concentration, suggesting a competition between PIH and ascorbate for complexation with Cu(lI). An almost full inhibitory effect by PIH was observed when the ratio PIH:copper was 1:1. A similar result was obtained with the measurement of copper plus ascorbate-mediated O2 uptake. Moreover, spectral studies of the copper and PIH interaction showed a peak at 455 nm and also indicated the formation of a stable Cu(II) complex with PIH with a 1:1 ratio. These data demonstrated that PIH prevents hydroxyl radical formation and oxidative damage to 2-deoxyribose by forming a complex with Cu(II) that is not reactive with ascorbate (first step of the reactions leading to hydroxyl radical formation from Cu(II), ascorbate and O2) and does not participate in Haber-Weiss reactions. PMID- 11855744 TI - Inhibition of Helicobacter pylori adherence by a peptide derived from neuraminyl lactose binding adhesin. AB - Helicobacterpylori, like many other gut colonizing bacteria, binds to sialic acid rich macromolecules present on the gastric epithelium. NLBH (neuraminyl lactose binding haemagglutinin) a 32 kDa adhesin located on the surface of H. pylori has been shown to have specific affinity towards NeuAcalpha2,3Galbeta1,4Gluc(3'SL). This sialic acid moiety is over-expressed in an atrophic stomach undergoing parietal cell depletion. Antibodies against a lysine rich peptide fragment of NLBH inhibit agglutination of human erythrocytes. This lysine rich sequence from NLBH was proposed to be the receptor-binding site. In order to elucidate the binding of NLBH to gastric epithelium, a peptide (D-P-K-R-T-I-Q-K-K-S) was synthesized. A series of experiments were performed involving adherence inhibition assays, 2D-NMR, molecular modelling and measurement of modulation in acid secretion. Results indicated that the peptide fragment could be involved in receptor recognition, which is important for the binding of H. pylori to gastric epithelium. The binding is possibly through hydrogen bonding. Two lysines and a threonine residue seem to be within the hydrogen bonding distance of NeuAcalpha2,3Galbeta1,4Gluc. Further, in vitro assays were performed to evaluate the role of the peptide on acid secretion by parietal cells isolated from human fundal biopsies. Interestingly, the peptide increases acid secretion only in H. pylori negative and in treated patients but not in H. pylori positive patients. This highlights the role of NLBH in acid secretion and could be of some consequence in the prognosis of the disease. PMID- 11855745 TI - Effect of the calcium phosphate-mediated RNA uptake on the transfer of cellular immunity of a synthetic peptide of HIV-1 to human lymphocytes by exogenous RNA. AB - It is known that exogenous RNA molecules can be taken up by eukaryotic cells and can exert a variety of biological effects both in vitro and in vivo. The modulation of human lymphocytes by exogenous RNAs has medical implications. The exogenous RNA used in this study was obtained from lymphoid organs of animals immunized with the synthetic peptide p12 of HIV-1 and was referred to as p12-RNA. Human lymphocytes were transfected with the p12-RNA and the transfer of immunoreactivity of p12 was assessed by the lymphocyte proliferation and the leukocyte adherence inhibition assays. Our results indicate that the transfer of cellular immune response to the p12 occurred in 9 donors (60%) who were named responsive individuals whereas 6 donors (40%) were non-responsives. We also found that the calcium phosphate-mediated RNA uptake method is effective in converting non-responsive into responsive donors. The calcium phosphate-mediated RNA uptake may also be used to increase the efficiency of RNA transfection in other models with medical implications and to contribute to a better understanding of the molecular events involved in the uptake of RNA. Our findings give support for the use of exogenous RNAs obtained from lymphoid organs of immunized animals with synthetic peptides of HIV-1 in the immune reconstitution of individuals infected with HIV-1. PMID- 11855746 TI - Differential effects of bacterial toxins on mitogenic actions of sodium fluoride and those of aluminum fluoride in human TE85 osteosarcoma cells. AB - This study compared the effects of cholera toxin (CTX) and pertussis toxin (PTX) on the actions of sodium fluoride (NaF) and those of aluminum fluoride (AlF3) on cell proliferation and differentiation, as well as tyrosine phosphorylation level of mitogen activated protein kinase (MAPK) in human bone cells. NaF and AlF3 each significantly stimulated the proliferation of human TE85 osteosarcoma cells, increased cellular alkaline phosphatase (ALP) activity, and increased MAPK tyrosine phosphorylation level. CTX completely blocked the bone cell anabolic activities of both NaF and AlF3. While PTX (2 ng/ml) inhibited the bone cell actions of NaF, it had no significant effect on those of AlF3. Both CTX and PTX completely blocked the stimulatory action of AlF3 on MAPK tyrosine phosphorylation, but neither toxin had an effect on the action of NaF on MAPK tyrosine phosphorylation. In conclusion, PTX and CTX had contrasting effects on the anabolic bone cell actions of NaF and AlF3 actions. These findings argue against the hypothesis that the osteogenic activity of NaF is mediated via the formation of AlF3 in human TE85 osteosarcoma cells. PMID- 11855747 TI - Differential expression of WISP-1 and WISP-2 genes in normal and transformed human breast cell lines. AB - The transcriptional alterations of specific gene(s) are actively associated with the development of different cancers including breast. The preceding studies of different laboratories documented at least 40 genes that may contribute directly to the genesis of cancer. Using differential display, RT-PCR and DNA sequencing analyses in normal human mammary epithelial cells (HMEC) and various breast tumor cell lines including MCF-7, ZR-75, T-47D and SKBR2, we demonstrated that WISP-1 and WISP-2 genes are differentially transcribed in these cells. WISP-2 mRNA transcription was identified in all 4 tumor derived cell lines, but the mRNA expression was undetected or minimally detected in normal breast epithelial cells. WISP-1 mRNA expression was identified in normal and transformed cell lines. However, the level of expression was higher in different breast tumor cell lines as compared to HMEC. The mRNA expression profiles of WISP genes in normal breast epithelial cells and breast tumor derived cell lines indicated a strong possibility of the involvement of WISP-signaling in the development of human breast tumors, and can be utilized as genetic markers of this disease. PMID- 11855749 TI - Oral activity and pharmacokinetics of 5,6-dimethylxanthenone-4-acetic acid (DMXAA) in mice. AB - PURPOSE: 5,6-Dimethylxanthenone-4-acetic acid (DMXAA), an anticancer drug with an antivascular action, has recently completed phase I clinical trials. Since oral administration has many advantages, we compared the biological activity and pharmacokinetics of DMXAA in mice following oral and intraperitoneal (i.p.) administration. METHODS: Growth delays of Colon 38 tumours were measured in C57Bl/6 mice. Plasma concentrations of DMXAA, 5-hydroxyindole-3-acetic acid (5HIAA) as a measure of serotonin production, and nitrate as a measure of nitric oxide production, were determined by high-performance liquid chromatography. Tumour necrosis factor (TNF) concentrations in serum and tumour tissues were measured by ELISA. RESULTS: The antitumour activity of DMXAA at the maximum tolerated oral dose (32.5 mg/kg) was low (4-day growth delay, no cures) compared to that (19-day growth delay, 40% cures) at the maximum tolerated i.p. dose (27.5 mg/kg). The pharmacokinetics of DMXAA in plasma, liver and tumour tissue indicated a bioavailability of 73%. Elevation of plasma 5HIAA, measured 4 h following i.p. administration of DMXAA, was linear with DMXAA dose, and the 5HIAA response to oral administration was consistent with its bioavailability. TNF concentrations increased following oral administration (30 mg/kg) and were particularly evident in tumour tissue, but were lower and less prolonged than those in response to i.p. administration at 25 mg/kg. Plasma nitrate levels were not increased following oral administration (30 mg/kg). CONCLUSIONS: DMXAA exhibits good bioavailability, and changes in serum TNF, tissue TNF, plasma 5HIAA and plasma nitrate, as markers of biological response, are consistent with this bioavailability. The low maximal plasma DMXAA concentration following oral administration, resulting in reduced retention of intratumoral TNF, may be responsible for the low antitumour activity. PMID- 11855748 TI - A study of amsalog (CI-921) administered orally on a 5-day schedule, with bioavailability and pharmacokinetically guided dose escalation. AB - PURPOSE: Amsalog is a derivative of 9-aminoacridine. Phase I studies using intravenous (i.v.) amsalog have shown the dose-limiting toxicity (DLT) to be phlebitis and myelosuppression. Phase II studies using a variety of schedules have shown evidence of activity in patients with large-cell lung, breast, and head and neck cancers. Preclinical studies demonstrated that amsalog is active orally: a clinical study of the oral bioavailability of amsalog was therefore performed. METHODS: A group of 20 patients with refractory malignancies were treated. There were two phases of the study: a pharmacokinetic comparison of i.v. against oral amsalog, followed by a pharmacokinetically guided oral dose escalation study. In the first phase of the study, 11 patients were treated. Amsalog 50 mg/m2 was administered i.v., and 50 mg/m2 and 200 mg/m2 orally. In the second phase of the study, 9 patients were treated in three cohorts of three. On day 1 of a 5-day schedule, amsalog was administered i.v. at the maximum tolerated dose (MTD) of 200 mg/m2. Subsequent doses were given orally, starting at a dose of 200 mg/m2 per day, with intrapatient dose escalation of up to 100% for the second cycle. Doses were escalated further in subsequent cohorts, based on oral bioavailability and toxicity. RESULTS: Oral bioavailability of 50 mg/m2 amsalog was 34%. In the dose escalation phase, DLT was neutropenia; other toxicities included malaise and nausea. The MTD was 1600 mg/m2 per day for 5 days. The plasma AUC using 1600 mg/m2 by the oral route was higher than that achieved using 200 mg/m2 by the i.v. route. CONCLUSION: Amsalog can be tolerated orally on a 5 day schedule at doses up to 1600 mg/m2. The recommended dose for further evaluation is 800 mg/m2 daily for 5 days, repeated three weekly. PMID- 11855750 TI - Phenylbutyrate-induced apoptosis is associated with inactivation of NF-kappaB IN HT-29 colon cancer cells. AB - PURPOSE: Cytotoxic chemotherapy has been used to treat patients with metastatic colorectal cancer with limited success. Therefore novel chemotherapeutic approaches are needed. Based on encouraging preclinical data, there has been an interest in developing derivatives of butyrate as clinically applicable agents. The purpose of this study was to investigate the effects of phenylbutyrate (PB), a butyrate analogue, on the cell growth and apoptosis in a colon cancer cell model. METHODS: Growth curves, flow cytometric studies, Western blotting, DNA binding assays and transient transfection experiments were performed in vitro using the colon cancer cell line HT-29 after exposure to PB. RESULTS: Exposure of HT-29 colon cancer cells to PB resulted in growth inhibition and induction of apoptosis as measured by annexin V staining. This increase in apoptosis was associated with a decrease in mitochondrial membrane potential, an increase in caspase-3 activity and a decrease in intact PARP protein levels. Since NF-kappaB plays a pivotal role in the regulation of apoptosis, we explored the effects of PB on the DNA binding and transcriptional activity of this transcription factor. After PB treatment, NF-kappaB-DNA binding was markedly decreased and specifically, this decreased DNA binding was observed in the p50:p65 heterodimer. The decreased NF-kappaB DNA binding was observed as early as 3 h after PB treatment, while no apparent changes in annexin V binding were detected until 12 h after PB treatment. Untreated HT-29 cells transfected with a kappaB-luciferase reporter plasmid demonstrated significant constitutive activity of the kappaB binding site, which was markedly decreased after treating the cells with PB. CONCLUSION: These results suggest that PB-induced apoptosis may be partly regulated through the inactivation of NF-kappaB. PB, an oral butyrate analogue, may have therapeutic potential in colon cancer. PMID- 11855751 TI - Enhanced antitumor activities of TZT-1027 against TNF-alpha or IL-6 secreting Lewis lung carcinoma in vivo. AB - PURPOSE: TZT-1027, an antimicrotubule agent that inhibits the polymerization of tubulin, shows potent antitumor activity in various transplantable tumor models in vivo. The high antitumor activity of TZT-1027 prompted us to speculate that this compound may have a mode of action other than its antimicrotubule and antimitotic activities. To elucidate the interaction of antitumor cytokines with TZT-1027 in tumors in vivo, we examined the antitumor activity of this agent against various cytokine gene-transfected Lewis lung carcinoma (LLC) cells inoculated into C57BL/6 mice. METHODS: In vitro growth inhibition was evaluated using the MTT assay, and in vivo activity was evaluated in subcutaneous models in C57BL/6 mice. The status of the vasculature in tumor tissues was evaluated immunohistochemically using anti-CD31 antibody. We used a cDNA macroarray to examine the gene expression profiles in tumor tissues removed from mice. RESULTS: TZT-1027 at 3 mg/kg showed potent antitumor activity in Mock (LLC-Neo cells) inoculated mice with a T/C% value of 16%. TZT-1027 at 3 mg/kg showed more potent antitumor activity in LLC-TNF cells and LLC-IL6 cells with T/C% values of 4% and 3%, respectively. TZT-1027 treatment destroyed the tumor vasculature as well as tumor cells in LLC-TNF and LLC-IL6 tissues of mice treated with TZT-1027. The LLC TNF and LLC-IL6 tissues of mice treated with TZT-1027 had in common the independent alteration of the non-histone chromosomal protein HMG-14 and transcription factor 1 for heat shock gene. Focusing on the gene regulation related to angiogenesis, the alteration in transcriptional factors such as ets family genes and homeobox family genes was remarkable. CONCLUSIONS: These factors are candidates as determinants of the enhanced TZT-1027 antitumor activity in relation to these cytokines. PMID- 11855752 TI - Blood and plasma pharmacokinetics of vinorelbine in elderly patients with advanced metastatic cancer. AB - PURPOSE: As vinorelbine is 78% bound to platelets, it seems interesting to investigate the pharmacokinetic profile of this drug from blood and to compare it to that determined from plasma. Thus, in this study, the comparative blood/plasma pharmacokinetics of vinorelbine were investigated in 15 elderly patients with advanced metastatic cancer. METHODS: The drug was given as a short (10 min) peripheral intravenous infusion; the administered dose ranged from 20 to 30 mg/m2 depending on the patient. Chemotherapy was repeated weekly. During the first and the fifth courses, each patient underwent pharmacokinetic evaluation. Toxicity evaluation was performed after each course of chemotherapy; a total of 109 courses was studied. Plasma and blood vinorelbine determinations were performed by high-performance liquid chromatography with spectrofluorimetric detection. Individual pharmacokinetic parameters were estimated with an empirical Bayes methodology. An open three-compartment pharmacokinetic model was used to describe the kinetics of vinorelbine. RESULTS: The half-lives of the terminal part of the curves, determined from blood and plasma data, were of the same order of magnitude: 31-35 h. Mean total clearances were about 0.71 l/h/kg from plasma and 0.45 l/h/kg from blood. Except during the first 15-20 min following the end of infusion, vinorelbine concentrations were 1.9 times higher in blood than in plasma. The ratio AUC(B)/AUC(P) (AUC(B) and AUC(P) are the area under the concentration-time curve from blood and plasma data, respectively) averaged 1.7; it was comparable to the blood/plasma ratio of 1.6 that remained constant over the 72 h of the study. There was substantial intra- and interpatient variability in vinorelbine pharmacokinetic parameters. This variability is similar within and between patients, and between pharmacokinetic parameters computed from blood and plasma. The elimination half-life is the parameter with the lowest intra- and interindividual variability (10-14%), while the AUC is the parameter presenting the highest variability (20-65%). The main haematological toxicity was anaemia (12 patients) and neutropenia (10 patients). Thrombocytopenia occurred in only one patient. At the first cycle, significant correlations were found between AUC(B) and AUC(P) and the decrease in neutrophil count (P < 0.05). The highest haematological toxicities encountered in this study occurred in patients presenting the lowest platelet count. AUC computed from plasma data decreased significantly with the increase of platelet count (P = 0.03). CONCLUSION: From the results of this study, blood did not appear to be a better predictor of haematological toxicity than plasma, but the decrease of platelet count seems to be a good predictor of this toxicity. Indeed, changes in platelet count are likely to produce strong variations in the unbound fraction of vinorelbine; this exposes the patient to a high risk of toxicity. PMID- 11855753 TI - Pharmacokinetics of Bcl-2 antisense oligonucleotide (G3139) combined with doxorubicin in SCID mice bearing human breast cancer solid tumor xenografts. AB - PURPOSE: To evaluate the pharmacokinetic (PK) properties of Bcl-2 antisense oligodeoxynucleotide G3139 when combined with the anthracycline anticancer drug doxorubicin (DOX) in a model of MDA435/LCC6 human breast cancer in severely compromised immunodeficient (SCID) mice. METHODS: An orthotopic model of MDA435/LCC6 solid breast tumors was developed by bilateral implantation of passaged cells in female SCID-RAG2 mice. The G3139 plasma profile was compared for two common routes of administration (i.v. or i.p.) in single and multiple dose treatment regimens of 5 mg/kg G3139 alone or with simultaneous DOX (5 mg/kg) administration. At selected times, plasma and major organs were assayed for [3H]G3139 using scintillation counting and DOX determined using HPLC. The molecular integrity of G3139 was analyzed using SDS-PAGE. The PKs of G3139 and DOX were evaluated using a two-compartment model. RESULTS: G3139 administered i.v. at 5 mg/kg revealed a biexponential plasma concentration-time curve with a Cmax of 99.9 microg/ml and elimination half-lives of 0.03 h and 9.8 h, respectively, which resulted in an area under the concentration-time curve (AUC) of 15.9 microg x h/ml. G3139 administered i.p. showed a plasma absorption, distribution and elimination profile typical of this route of administration, characterized by half-lives of 0.03 h, 0.2 h and 8.9 h, respectively and a Cmax of 8.6 microg/ml. Based on AUC comparisons, the bioavailability of G3139 injected i.p. was 84% compared to i.v. administration. Subtle changes were observed in G3139 PKs after three prior i.p. doses of G3139. Specifically, a six-fold slower absorption rate, lower Cmax (6.9 microg/ml), increased Tmax (0.2 h), and an AUC of 17.4 microg x h/ml were observed, consistent with concentrations approaching saturation levels in tissue sites to which G3139 distributes. Coadministration of DOX had significant effects on the PK properties of G3139, manifested by an increased Cmax (11.2 microg/ml), higher AUC (19.7 microg x h/ml), and ninefold lower plasma clearance for single-dose G3139 administration. G3139 in plasma remained largely intact (< 17% degraded in plasma over 4 h), and increased plasma protein association occurred as a function of time. G3139 was detected in both healthy and tumor tissue after i.v. and i.p. administration. The highest tissue levels of G3139 were observed in the kidneys (40 microg/g), and low levels (< 2 microg/g) were detected in lung, heart and muscle. The rate of accumulation of G3139 in organs was dependent upon G3139 levels in plasma and the presence of coadministered DOX. Significant accumulation of G3139 was observed in solid tumors, with peak levels of approximately 5 microg G3139/g tumor, and approximately a two-to threefold tumor/muscle AUC ratio. The kinetics of G3139 accumulation in tumor tissue increased with increasing circulating G3139 concentration. The tissue distribution properties of DOX were also altered in the presence of coadministered G3139: in the presence of G3139, tumor exposure to DOX increased two-to threefold without alteration in plasma DOX PKs. CONCLUSIONS: These findings indicate that drug-drug interactions between G3139 and DOX are modest and favorable in that elevated tumor DOX levels are achieved without compromising G3139 tumor uptake or significantly altering plasma drug concentrations. PMID- 11855754 TI - Antiangiogenic effects of a protein kinase Cbeta-selective small molecule. AB - BACKGROUND: Protein kinase C frequently plays a central role in the intracellular signal transduction of growth factors and cytokines. METHODS: The acyclic bisindolylmaleimide 317615 x 2HCl was identified as a potent selective inhibitor of protein kinase Cbeta. The compound 317615 x 2HCl was tested in culture and in vivo in the rat corneal micropocket and in the SW2 small-cell lung carcinoma human tumor xenograft. RESULTS: In cell culture, 317615 x 2HCl was a more potent inhibitor of VEGF-stimulated HUVEC proliferation (IC50 150 nM, 72 h) than of human SW2 small-cell lung carcinoma cell proliferation (IC50 3.5 microM, 72 h). When administered orally twice daily for 10 days, the compound 317615 x 2HCl markedly decreased the neoangiogenesis induced by VEGF or bFGF in the rat corneal micropocket assay. To assess antitumor efficacy, 317615 x 2HCl was administered orally twice daily to nude mice bearing SW2 xenograft tumors on days 14 through 30 after tumor implantation. The number of countable intratumoral vessels was decreased in a dose-dependent manner reaching as low as one-quarter the number in the control tumors. The decrease in intratumoral vessels was paralleled by increases in tumor growth delay. Treatment of the tumor-bearing animals with paclitaxel or carboplatin followed by treatment with 317615 x 2HCl resulted in a 2.5- to 3.0-fold increase in tumor growth delay compared with the standard chemotherapeutic agents alone. CONCLUSIONS: 317615 x 2HCl represents a new approach to antiangiogenic therapy in cancer-blocking multiple growth factor signaling pathways in endothelial cells with a single agent. 317615 x HCl is in early clinical testing. PMID- 11855755 TI - Pharmacokinetics, tissue distribution, and metabolism of 17 (dimethylaminoethylamino)-17-demethoxygeldanamycin (NSC 707545) in CD2F1 mice and Fischer 344 rats. AB - PURPOSE: 17-(Dimethylaminoethylamino)-17-demethoxygeldanamycin (17DMAG) is an analogue of the benzoquinone ansamycin compound 17-(allylamino)-17 demethoxygeldanamycin (17AAG), which is currently being evaluated in clinical trials. Studies were performed in mice and rats to: (1) define the plasma pharmacokinetics, tissue distribution, and urinary excretion of 17DMAG after i.v. delivery; (2) define the bioavailability of 17DMAG after i.p. and oral delivery; (3) characterize the biliary excretion of 17DMAG after i.v. delivery to rats; and (4) characterize, if possible, any metabolites of 17DMAG observed in plasma, tissue, urine, or bile. MATERIALS AND METHODS: Studies were performed in female, CD2F1 mice or male Fischer 344 rats. In preliminary toxicity studies and subsequent i.v. pharmacokinetic studies in mice, 17DMAG i.v. bolus doses of 33.3, 50, and 75 mg/kg were used. In bioavailability studies, i.p. and oral 17DMAG doses of 75 mg/kg were used. In preliminary toxicity studies in rats, i.v. bolus doses of 10 and 20 mg/kg were used, and in i.v. pharmacokinetic studies 10 mg/kg was used. Compartmental and noncompartmental analyses were applied to the plasma concentration versus time data. In mice and rats, concentrations of 17DMAG were determined in multiple tissues. Urine was collected from mice and rats treated with each of the i.v. doses of 17DMAG mentioned above, and drug excretion was calculated until 24 h after treatment. Biliary excretion of 17DMAG and metabolites was studied in bile duct-cannulated rats given a 10 mg/kg i.v. bolus dose of 17DMAG. 17DMAG metabolites were identified with LC/MS. RESULTS: A 75 mg/kg dose of 17DMAG caused no changes in appearance, appetite, waste elimination, or survival of treated mice as compared to vehicle-treated controls. Bolus i.v. delivery of 17DMAG at 75 mg/kg produced "peak" plasma 17DMAG concentrations between 18 and 24.2 microg/ml in mice killed at 5 min after injection. Sequential reduction in the 17DMAG dose to 50 and 33.3 mg/kg resulted in "peak" plasma 17DMAG concentrations between 9.4 and 14.4, and 8.4 and 10.5 microg/ml, respectively. Plasma 17DMAG AUC increased from 362 to 674 and 1150 microg/ml x min when the 17DMAG dose increased from 33.3 to 50 and 75 mg/kg, respectively, corresponding to a decrease in 17DMAG CLtb from 92 ml/min per kg to 75 and 65 ml/min per kg. Plasma 17DMAG concentration versus time data were best fit by a two-compartment open linear model. No potential 17DMAG metabolites were observed in plasma. 17DMAG bioavailability was 100% and 50% after i.p. and oral delivery, respectively. In rats, an i.v. bolus dose of 10 mg/kg produced peak plasma 17DMAG concentrations between 0.88 and 1.74 microg/ml. Plasma 17DMAG concentrations had fallen below the lower limit of quantitation by 180 min and were best fit by a one-compartment open linear model. The plasma 17DMAG AUC was 104 microg/ml x min, corresponding to a 17DMAG CLtb of 96 ml/min per kg. 17DMAG distributed rapidly to all mouse and rat tissues except brain and testes. Only mouse liver contained materials consistent with potential metabolites of 17DMAG, but their concentrations were below the limit of quantitation of the HPLC assay used. Within the first 24 h after delivery, urinary excretion of 17DMAG by mice and rats accounted for 10.6-14.8% and 12.5-16%, respectively, of the delivered dose. By 15 min after i.v. delivery of 10 mg/kg of 17DMAG, rat bile contained 11 new materials with absorbance similar to that of 17DMAG. Four of these proposed metabolites had an Mr of 633, indicating addition of an oxygen. Two of these proposed metabolites had an Mr of 603, implying the loss of one methyl group, and one had an Mr of 589, implying the loss of two methyl groups. The remaining four proposed metabolites had an Mr of 566, 571, 629, and 645, respectively. Biliary excretion of 17DMAG and metabolites accounted for 4.7 +/- 1.4% of the delivered dose, with 17DMAG accounting for 50.7 +/- 3.4% of the biliary excretion. CONCLUSIONS: 17DMAG has excellent bioavailability when given i.p. and good bioavailability when given orally. 17DMAG is widely distributed to tissues and is quantitatively metabolized much less than is 17AAG. The pharmacokinetic and metabolite data generated should prove relevant to the design of additional preclinical studies as well as to contemplated clinical trials of 17DMAG and could be useful in their interpretation. PMID- 11855756 TI - Doxorubicin-induced apoptosis and chemosensitivity in hepatoma cell lines. AB - PURPOSE: Doxorubicin (DOX) is a commonly used anticancer drug which causes DNA damage and kills cancer cells mainly by apoptosis. However, the process leading to killing of cancer cells and the molecular basis of resistance to DOX are not well understood. To evaluate the role of p53 and the cellular effects of DOX on hepatoma cell lines, we examined three hepatoma cell lines with different p53 status--Huh-7 (mutated p53), HepG2 (wild-type p53) and Hep3B (deleted p53). METHODS: The chemosensitivity of the three hepatoma cell lines was assessed using the MTT assay, and cell cycle distribution was evaluated by flow cytometry. Western blotting and immunostaining were employed to examine the protein alterations in response to DOX treatment, and a DNA fragmentation assay was performed to detect apoptosis. RESULTS: Of the three cell lines, HepG2 was found to be most resistant to DOX, followed by Hep3B, and Huh-7 was most sensitive to DOX treatment. HepG2 showed G1 arrest 24 h after drug administration and upregulation of p53 protein level in a time-dependent manner. In Hep3B cells (deleted p53), G2/M phase arrest was observed soon after drug administration, accompanied by induced apoptosis that was p53-independent. In Huh-7 cells (mutated p53), which were most sensitive to DOX, there was neither G1 nor G2 arrest, and the level of p53 mutated protein was downregulated after DOX treatment. MDM2 and p27 proteins were downregulated in all cell lines independently of p53 status. p21 was upregulated following p53 activation at low doses of DOX in HepG2 cells, but at higher doses, p21 was downregulated in Huh-7 and HepG2 cells. CONCLUSION: DOX confers different chemosensitivity on different hepatoma cell lines with different p53 status. The contrasting relationships between chemosensitivity and p53 status and expression suggest that DOX-induced apoptosis and cell death involve pathways that are independent of p53. PMID- 11855758 TI - Managing change. PMID- 11855757 TI - Atypical pharmacokinetics and excretion of new platinum analogues in rodents. AB - PURPOSE: Two new series of platinum complexes with cytotoxic activity in vivo are [Pt(NRCH2)2L2], (R=polyfluorophenyl, L=pyridine or substituted pyridine) and [Pt(NRCH2CH2NR'2)2L(X)], (R, L as before; R'=Me or Et, X=halogen). The aim of this study was to determine the pharmacokinetics and excretion in mice and in isolated perfused rat livers of a representative compound from each class, respectively: Pt103 (R=p-HC6F4, L=pyridine) and Pt109 (R, L as for Pt103, R'=Et, X=I). METHODS: Mice were given intraperitoneal injections of active doses of Pt103, Pt109, or cisplatin in a variety of vehicles. Blood was sampled at several times to 48 h. Some mice were placed in metabolic cages where urine and feces were collected. In isolated, perfused rat livers, perfusate and bile were collected following a dose of Pt103, cisplatin or carboplatin. Platinum was measured in blood, urine, feces, or perfusate by atomic absorption spectroscopy. Three vehicles used were peanut oil, dimethyl sulphoxide, and saline/Tween 20. RESULTS: In contrast to renal excretion of over 70% for cisplatin in saline, urinary excretion of platinum was less than 24% of a dose of Pt109 in peanut oil, less than 21% of P103 in DMSO, and only 4% for Pt103 in peanut oil. Over 60% of Pt103 was eliminated in mouse feces, and 57% was excreted in bile from rat liver. Plasma protein binding of Pt109 was greater than 90% at 6 h following administration in mice. CONCLUSION: In contrast to cisplatin and carboplatin, representatives of two new classes of platinum anticancer agents undergo minimal renal elimination, but are excreted mainly in the bile and feces. If a platinum complex with a similar excretion profile was introduced into the clinic, there might be a therapeutic advantage in terms of drug toxicity and combination therapy with other cytotoxics. PMID- 11855759 TI - Deroofing minor burn blisters--what is the evidence? AB - It is estimated that 150000 new patients per year will attend an A&E Department or Minor Injury Unit (MIU) with a minor burn injury (Wardrope & Edhouse 1999). There is often conflicting 'evidence' for the management and treatment of minor burns blisters and this can cause variations in the treatment provided within an A&E department resulting in confusion for staff and patients. PMID- 11855760 TI - Dilemma--volunteering help. AB - Last Saturday afternoon I was playing with my family on the beach of a British seaside resort. It became clear that an incident had occurred in the water, when the beach patrol drove very quickly onto the sands closely followed by police cars. The policemen ran straight into the water to pull a small speedboat up on to the beach. I considered joining them to see if any assistance was required, but felt slightly uncomfortable about pushing myself forward, despite being trained in the management of patients with major injuries. A couple of minutes later, two paramedics arrived in an ambulance. Everyone seemed to converge on the small speedboat and I decided to approach the scene in case assistance was required to lift the casualty. When I approached the side of the boat I saw a great deal of blood everywhere and 8 or 10 people leaning over the patient. I asked 'Can I help?' There was no reply. I looked at a woman and said 'Do you need me or do you know what you are doing?' The lady replied to me 'No, I think it's okay, there's a doctor and two paramedics.' I backed off immediately, concerned about treading on people's toes. Twenty minutes later a police helicopter arrived at the scene and the patient had still not been taken out of the boat. Ten minutes after the helicopter had arrived and the patient had still not been moved, I once again, approached the scene and spoke to a police officer. 'I am trained to lift and turn patients with potential spinal injuries. Do you think they need any assistance?' The young policewoman went off to talk to a more senior policeman nearer to the incident, however, she did not speak to the paramedics. After five minutes, I approached both policemen again, and asked if I was required. The policemen both said no and waved me away. It was a further fifteen minutes before the patient was removed on the spine board from the boat, and put into the helicopter. I remained uncomfortable for the rest of the weekend about my response. I realize I tried hard not to offend other health care professionals at the scene, but am anxious that in doing so, I jeopardized the safety of the patient. I have no idea what training the medical bystander had or whether the paramedics were pre-hospital trauma life support trained. Should I have made more effort to satisfy myself that the most appropriate people looked after the patient? PMID- 11855761 TI - Lynn Sbaih interviews Chris Moulton, A&E consultant to the Royal Bolton Hospitals NHS Trust, Bolton, UK. PMID- 11855762 TI - Current triage practice and influences affecting clinical decision-making in emergency departments in NSW, Australia. AB - In Australia, as elsewhere, the nature of triage decision making, patient referral, investigations, physical resources, triage policies, educational requirements and clinical expertise is often unclear and differs between organizations (Gerdtz & Bucknell 2000; Standen 1998). The study described here was undertaken in order to explore current triage practices throughout New South Wales (NSW) and to describe the range of clinical reasoning tasks performed by nurses. PMID- 11855763 TI - Using the Delphi technique to design a self-reporting triage survey tool. AB - A useful addition to any researcher's methodological repertoire is the Delphi technique. This technique facilitates group communication to develop research items that are contextually specific. For the first time this method was utilised to develop a self-reporting triage survey tool that would explore current practice and begin to explore triage decision-making. The triage role is complex, often crisis-driven and demands speed and accuracy on the part of the triage nurse. This role involves patient assessment, initiating investigations, patient referral and management. The study used the Delphi technique to produce a triage survey tool that would assist in broadening our understanding of current triage practice within Australia. PMID- 11855764 TI - Unexpected birth in A&E departments. AB - This paper discusses the current advice on and management of a woman who gives birth unexpectedly. It is written from the perspective of a midwife teacher who, whilst having fifteen years of midwifery experience, has not practised within an A&E department. However, lectures on the subject of unexpected birth to students undertaking trauma modules have led me to the conclusion that this event provokes anxiety in even the most experienced staff. Reflection upon their practice and discussion also leads them to appreciate that there are many invaluable skills, which are already part of every trauma nurse's repertoire and which are transferable to the situation of unexpected birth. An outline of the occasions when women may present unexpectedly in labour will be offered, followed by an outline of the physiology of normal labour and suggested management. This paper is not intended to replate departmental protocols; rather it offers some useful and basic information for guidance and will hopefully allay some anxieties regarding the skills required to effectively assist a woman to give birth. PMID- 11855765 TI - Scaphoid injuries in children. AB - In order to establish an acceptable working protocol for the care of children with scaphoid injuries, an independent study was undertaken by the author. This paper describes the various aspects of this study: understanding what the scaphoid is and its function, the correct examination of the hand and wrist and the rationale behind it, the rationale behind the current treatment of suspected scaphoid injuries, and the development of the carpal bones and skeletal age. Opinions differ as to what age the scaphoid develops and therefore when it may be fractured, and so a review of research and literature on scaphoid injuries is also outlined. PMID- 11855766 TI - Male catheterization. AB - The insertion of catheters into male emergency patients is fairly common practice and is associated with a worryingly high rate of infection. Everyday pressures within the department, along with the added stress of resuscitation can result in inappropriately trained or skilled staff undertaking this procedure. The issue of gender and whether female nurses should catheterize male patients may also affect this vulnerable group of patients. Acquiring the psychomotor skills of inserting a urethral catheter is only one part of preparation for practice. Emergency nurses must know when and when not to resort to catheterization. Choosing the type and size of catheter requires careful judgment. What will you do if insertion proves difficult? Prevention of infection is of paramount importance and there are an increasing number of evidence-based sources of information, which are crucial to formulating procedures and informing every day practice. In the pressured surroundings of A&E departments, it is easy to ignore the vulnerability of men requiring catheterization, both from a physical and psychological point of view. Making the effort to explain the procedure, listen to questions and concerns and record relevant details in the notes, will take only a few extra moments. There is no doubt that urinary catheterization is not without complications. It is associated with significant morbidity and occasionally, mortality. PMID- 11855767 TI - Shaping the future: reforming routine emergency nursing work. AB - The provision of emergency care, for many A&E nurses, is dependent upon taken-for granted assumptions, associated with the ongoing management of the changing shape of the work. In particular, routine working practices and procedures reflect a collective ideology of A&E nursing labour which involves moving people on and out of the emergency department. Subscribing to this ideology and realizing good shape provides nurses with a feeling that they are in control of their work and have done a good job. Analysis of the NHS Plan has identified that the Government's goals for reform support as well as challenge the ways in which nurses currently strive towards maintaining the good shape of their work. In particular, the ideology of patient-centredness and the Chief Nursing Officer's ten key roles for nurses, provide an opportunity for nurses to explore the ways in which they can cultivate their routine activities and contribute to the shape of future emergency nursing work. PMID- 11855768 TI - An unusual case. PMID- 11855769 TI - 'Aroma 2000--fragrancing the future'. Favre Armstrong reports on the first Baltic States Aromatherapy Conference. PMID- 11855770 TI - A little piece of history...views of acute pain management in paediatric oncology. PMID- 11855772 TI - How might traditional remedies be incorporated into discussions of integrated medicine? PMID- 11855771 TI - Integrated Medicine Conference Report: can alternative medicine be integrated into mainstream care? PMID- 11855773 TI - Relaxation training as complementary therapy for mild hypertension control and the implications of evidence-based medicine. AB - Relaxation therapy for the treatment of hypertension presented a number of challenges, in terms of conducting and using research as well as gaining the wider acceptance of this complementary therapy in common practice in Hong Kong. Two issues were central to the current implementation of complementary therapies: the evidence-based practice movement and the management of risk during the implementation of new therapies. A major concern was how to maximize the chance of inclusion of research on complementary therapies in evidence-based practice systematic reviews. This was thought to be achievable by making the data amenable to meta-analysis when small samples may be unavoidable. The empirical work examined the effects of three relaxation therapies for the reduction of high blood pressure in nine Chinese subjects. Subjects were randomly assigned to three groups: (a) progressive muscle relaxation, (b) stretch release relaxation and (c) cognitive imagery relaxation. Systolic, diastolic blood pressure and heart rate were assessed in a baseline session, the 8th post-treatment session, and a 30 days follow-up session. Datawere analyzed using ANOVA and Paired sample t-test. One-Sample Kolmogorov-Smirnov Tests for the normal distribution were performed among the three groups. The results verified that the normality assumption for the data had not been violated. Results revealed that in the context of the study all relaxation therapies can reduce blood pressure in Chinese subjects, but stretch release relaxation and progressive muscle relaxation therapies appeared to be more effective in lowering blood pressure compared to cognitive imagery relaxation. These data are in a form available for meta-analysis at a later date. The study also shows a degree of clinical significance and validation with respect to a Hong Kong population. It demonstrates ways in which the inclusion of research on complementary therapies in systematic reviews can be maximized. PMID- 11855775 TI - A comparison of the use of complementary therapies in Australia and Iceland. AB - The use of complementary therapies in Iceland and Australia is compared in this paper, along with information regarding research into native plants in both countries. Attitudes towards complementary therapists from the medical profession are discussed, and the use of complementary medicine by pregnant women and midwives is given special emphasis. PMID- 11855774 TI - Brain injury: the uncertainties of using complementary therapies. AB - The use of complementary therapies is fast growing in the UK, but their place within health care is still unclear. This study explored the views of families using a specific complementary therapy in the care of their brain-injured children, and of professionals involved in the care of the children. The findings revealed an interesting comparison of views about the use of complementary therapies and attitudes towards their use. PMID- 11855776 TI - Aromatherapy massage for joint pain and constipation in a patient with Guillian Barre. AB - The following case study will look at the efficacy of aromatherapy massage in a patient diagnosed with Guillian Barre Syndrome admitted to an IntensiveTherapy Unit. The pathophysiology of this disorder will be discussed, medical treatment will be outlined and adjuncts to conventional nursing care will be presented. Aromatherapy massage was used to complement the conventional nursing and medical treatment of joint pain and constipation. The Mead Model for nursing care was used for assessment and the plan of care devised from this. Evaluation of outcomes were incorporated into the implementation protocol to ensure positive outcomes were achieved. PMID- 11855777 TI - Touch Research Institutes: an interview with Dr Tiffany Field (interview by Peter Mackereth). AB - The following paper reports on a recent visit to the Touch Research Institutes (TRI) based at the University of Miami School of Medicine. The main purpose of making the journey was to participate in a 3-day Research Training Workshop. The paper will first provide a brief overview of the history and work of the TRI. There will then be an interview with Dr Tiffany Field, the TRI founder and Director, and finally accounts of the workshop experience itself and a visit to one of Miami's foremost schools of massage therapy: 'Educating Hands'. PMID- 11855778 TI - Complementary therapies courses for the 21st century. AB - Over the past 5 years, an innovative portfolio of degrees in complementary therapies has been developed at the University of Westminster. Most of the degrees were the first of their kind in the UK, and the provision is now the widest range of complementary therapies courses in Europe. With an intake of 80 students in 1996, the provision has grown rapidly to a total of over 500 in 2000. The opening of the unique multidisciplinary Polyclinic in 1998 marked the beginning of an exciting clinical training and research dimension. The context that has lead to the growth of mainstream higher education provision of complementary therapies is explored. The current offer is reviewed, and criteria for degree provision and the role of the professional bodies discussed. The development of the University of Westminster's provision is described, including the early collaboration with established providers, design, influence of nursing and midwifery, experiences with the professional bodies, development of the unique Polyclinic and the plans for the future. PMID- 11855779 TI - An interview with Alison Demarco and Nichol Clarke: light and colour therapy explained. PMID- 11855780 TI - Azurophil granules are heterogeneous with respect to mobilization induced by different concentrations of fMLP. AB - Azurophil granules of neutrophils beyond their already known heterogeneity of beta-glucuronidase and myeloperoxidase enzyme contents are heterogeneous with respect to a spontaneous or low concentration (2.3 or 23 nM) of formyl-Met-Leu Phe-induced mobilization. This suggests that the heterogeneity of azurophil granules is manifested in their functions too. PMID- 11855781 TI - Induction of apoptosis and bcl-2 expression in acute lymphoblastic leukaemia and non-Hodgkin's lymphoma in children. AB - bcl-2 expression is associated with the expression of the multidrug resistance molecule (p-gp) and the resistance of leukaemia cells to the induction of apoptosis. The activity of p-gp is the main mechanism of resistance of leukaemia cells to chemotherapy. This study assessed the induction of apoptosis of acute lymphoblastic leukaemia (ALL) and non-Hodgkin's lymphoma (NHL) blastic cells following in vitro treatment with dexamethasone (DXM), vincristine (VCR), and tumour necrosis factor (TNF) in relation to the expression of bcl-2 and p-gp. Common ALL (cALL; n = 24 patients), common ALL with co-expression of myeloid antigens (cALL + My; n = 9), ALL-T (n = 9), and NHL [n = 6 (T type, n = 2; B type, n = 4)] were included. The expression of bcl-2 and p-gp and apoptosis were assayed by flow cytometry. Spontaneous apoptosis was low (< 5%) in cALL and ALL-T and higher (> 8%) in NHL and cALL + My. A high frequency of bcl-2 expression was noted in cALL and cALL + My. A high frequency of p-gp expression was observed in cALL + My, ALL-T, and NHL. There was a reverse association between bcl-2 expression and spontaneous apoptosis. DXM-induced apoptosis was observed in 52.63%, TNF-induced in 42.85%, VCR-induced in 36.36%, and GM-CSF-induced in 33.3% of leukaemia and lymphoma cases. DXM and GM-CSF-driven apoptosis was reversibly associated with bcl-2-expression (bcl-2-dependent mechanism). VCR and TNF-driven apoptosis was not associated with bcl-2 expression, suggesting a different, bcl-2 independent, mechanism(s) of its induction. The in vitro induction of apoptosis was not associated with expression of p-gp. PMID- 11855782 TI - Acute lymphoblastic leukemia accompanied by chromosomal abnormality of translocation (12;17). AB - This report concerns a female patient with B-precursor acute lymphoblastic leukemia with concomitant expression of CD13 and CD33. A rare chromosomal abnormality, t(2;17)(p13:q21), was detected in 70% of metaphases analyzed. A new subgroup of acute leukemia is proposed, consisting of our case and 12 reported cases with similar chromosomal abnormalities. This subgroup has a poor prognosis and a high incidence of biphenotypic characteristics. PMID- 11855783 TI - Metastatic bone marrow tumours: a report of six cases and review of the literature. AB - Bone marrow aspirations and biopsies are now employed in the investigation of many disorders in haematology, oncology, and internal medicine. In this paper, clinical, laboratory, and pathological findings are presented of patients with bone marrow metastases of solid tumours (n = 6). Anemia was detected in four cases (66.6%), thrombocytopenia in five cases (83.3%), leukopenia in three cases (50%), diffuse bone pain in four cases (66.6%), elevated serum alkaline phosphatase levels in four cases (66.6%). and hypercalcaemia in two cases (33.3%). In conclusion, bone marrow aspiration and biopsy examinations are effective, practical, and cheap in detecting solid tumour metastasis to bone marrow in a selected group of patients. PMID- 11855784 TI - Mixed cryoglobulinaemia and B-cell lymphoma in the absence of hepatitis C virus infection. AB - Literary data favouring hepatitis C virus infection as an important aetiological agent in mixed cryoglobulinaemia and mixed cryoglobulinaemia-associated lymphoma have been reported. Hepatitis C virus infection does not explain, however, all cases. A case of hepatitic C negative mixed cryoglobulinaemia with B-cell lymphoma is presented and the possible role of hepatitis C virus in the aetiology of lymphoproliferative disorders and the geographical differences reported in the literature are discussed. PMID- 11855785 TI - Analysis of IL-18 bioactivity and IL-18 mRNA in three patients with adult T-cell leukaemia, acute mixed lineage leukaemia, and acute lymphocytic leukaemia accompanied with high serum IL-18 levels. AB - Interleukin-18 (IL-18) bioactivity in sera and IL-18 mRNA expression in leukaemia cells of three patients with adult T-cell leukaemia (ATL), acute mixed lineage leukaemia (AMLL) and acute lymphocytic leukaemia (ALL) accompanied with high serum IL-18 levels have been analysed. There was little serum IL-18 bioactivity in the three patients with ATL, AMLL and ALL, while IL-18 mRNA expression was detected in leukaemia cells of all three patients. PMID- 11855786 TI - Increased plasma levels of interleukin-6 and interleukin-8 in beta-thalassaemia major. AB - Several immunological defects can be found in patients with beta-thalassaemia, among which the impairment of neurophil and macrophage phagocytic and killing functions and the production of some cytokines are the most important. It is known that interleukin-6 (IL-6) and interleukin-8 (IL-8) are important components of the pro-inflammatory response. The plasma levels of these cytokines may be relevant in the pathophysiology of beta-thalassaemia. To assess this hypothesis, the plasma IL-6 and IL-8 concentrations in patients with beta-thalassaemia, were investigated. Fourteen patients with thalassaemia major were studied by evaluating body iron status, iron supply for erythropoiesis, and plasma IL-6 and IL-8 levels, together with 12 age-matched healthy controls. The plasma levels of IL-6 and IL-8 were determined by enzyme-linked immunosorbent assay (ELISA). Patients with beta-thalassaemia were found to have higher IL-8 concentrations than normal controls (p < 0.001) and plasma IL-6 concentrations increased significantly in the beta-thalassaemic patients compared with control subjects (p = 0.01). Serum ferritin levels of beta-thalassaemic patients were significantly higher than those of control groups (p < 0.05). IL-8 levels correlated with ferritin levels (r = 0.694; p < 0.05) and the total number of transfusions (r = 0.64; p < 0.05). Plasma IL-6 levels in beta-thalassaemic patients did not correlate with any clinical, haematological or biochemical parameters. It was also found that plasma IL-8 levels in the patients who had blood transfusions over 100 times were significantly higher than those of under 100 times (p < 0.05), whereas there was no statistical difference for IL-6. Markedly increased plasma IL-6 and IL-8 levels were documented in patients with beta-thalassaemia. Increased production of IL-6 and IL-8 might have contributed to abnormalities in iron metabolism and it is probably due to overstimulation of macrophages. Before a clinical value can be ascribed to these changes in plasma cytokine levels in beta-thalassaemia, the follow-up samples of larger series of patients with 8 thalassaemia should be evaluated. PMID- 11855787 TI - Does amphotericin B lipid complex (Abelcet) affect haematological parameters erroneously? AB - Abelcet is composed of large particles which could be misinterpreted as blood cells on measurements with blood cell counters. The direct measurement of drug suspensions on blood cell counters has been performed in an in vitro study. In an ex vivo study, the haematological parameters were compared before and during Abelcet infusions. A significant interference effect was observed in platelet counts, together with minimal differences in the WBC, RBC, and Hb parameters in the in vitro study. In the ex vivo study, there were statistically significant deviations only in RBC counts and in haemoglobin (Hb) level, while there was no difference in the other parameters. It is been reported that the drug accumulates very rapidly in the reticuloendothelial system and circulates minimally in the plasma. That is why there is a significant deviation in the direct counting of platelets, while the platelet counts taken from the patients do not differ statistically. PMID- 11855788 TI - Seroprevalence of HBsAg, anti-HCV, and anti-HIV among haemophiliac patients in Shiraz, Iran. AB - A study was performed during 1999-2000 on multi-transfused patients with haemophilia who are registered by the Shiraz Haemophilia Society. HBsAg, anti HCV, and anti-HIV were checked using a second-generation enzyme-linked immunosorbent assay (ELISA). Positive tests for anti-HCV and anti-HIV were confirmed by a western blot test. Healthy blood donors were used for the control group. HBsAg, anti-HCV, and anti-HIV were positive in two (0.71%, 95% CI = 0.12 2.33), 44 (15.65%, 95% CI = 11.76-20.26), and one (0.36%, 95% CI = 0.02-1.74) of the patients, respectively. Positive sera for HBsAg, anti-HCV, and anti-HIV were found in 85 (1.07%), 47 (0.59%), and 27 (0.34%) of the control group, respectively. The rate of anti-HCV was significantly higher in the patients than in the control group (p << 0.0001). The rate of positive anti-HCV was significantly higher than that of positive HBsAg in the patients (p << 0.0001). The reverse was correct for the control group (p = 0.0008). It is concluded that HCV is the current major problem in multi-transfused haemophiliac patients and more careful pre-transfusion screening of blood for anti-HCV must be introduced in all blood banks. PMID- 11855789 TI - HLA immunization of haemodialysed patients in the transfusion and the erythropoietin eras. AB - The frequency of transfusions and the development of HLA alloimmunization in the years 1986 and 1995, representing the 'transfusion era' and the 'erythropoietin era', have been compared. The introduction of erythropoietin was found to reduce the need for transfusion in haemodialysed patients on the waiting list from 65.2% to 5.7%. The elimination of transfusions resulted in a decrease in HLA immunization from 56.5% to 35%. In those patients who were still immunized, antibody production was probably caused either by immunomodulation or by transfusions given to treat complications. Erythropoietin could be used instead of transfusions in almost all cases, except for complications involving severe bleeding. PMID- 11855790 TI - Treatment of steroid-resistant idiopathic thrombocytopenic purpura in pregnancy with repeated high-dose intravenous immunoglobulin. AB - High-dose intravenous immunoglobulin (HD-IVIG) has temporary but reliable efficacy in idiopathic thrombocytopenic purpura (ITP). HD-IVIG has been described as the representative management in pregnant cases of refractory to corticosteroid or immunosuppressants. There have been few cases treated with repeated HD-IVIG to sustain pregnancy from the early phase of pregnancy. This case report describes a pregnant case of steroid-refractory ITP, treated with six times repeated HD-IVIG, resulting in the successful delivery of a healthy newborn with a normal platelet count. No adverse effects were observed. PMID- 11855791 TI - Urinary trypsin inhibitor levels in the urine of patients with haematological malignancies. AB - The urinary trypsin inhibitor (UTI) levels in the urine of patients with various haematological malignancies were determined, using automated latex agglutination immunoturbidimetry. The mean UTI levels in urine in acute non-lymphocytic leukaemia, myelodysplastic syndrome, non-Hodgkin's lymphoma, and multiple myeloma groups were significantly elevated, compared with the normal control group. It was found that the UTI level in urine changed from an elevated value to a normal value with haematological improvement by chemotherapy in a patient with myelodysplastic syndrome included in a previous study. These results suggest tha PMID- 11855792 TI - Lack of an association of urinary albumin excretion with interleukin-6 or C reactive protein in patients with type 2 diabetes. AB - The reason for the elevation of fibrinogen concentration in diabetic patients with nephropathy is not known so far. In order to elucidate the mechanism of such an increase in fibrinogen levels, we investigated haemorheological and inflammatory markers in type 2 diabetic patients in a cross-sectional design. Thirty-two non-smoking type 2 diabetic patients (13 women, 19 men; body mass index 29.1+/-5.4 kg/m2, age 62.8+/-12.1 years) were investigated. Patients with C reactive protein levels >1.5 mg/dl were excluded from the study. Concentration of fibrinogen was measured by immunonephelometry, C-reactive protein by immunoturbidimetry, and interleukin-6 (IL-6) by an enzyme-linked immunosorbent assay, and viscosity of plasma and of whole blood was determined by rotation viscosimetry. Concentrations of inflammatory parameters were well correlated with each other (p<0.05 for all correlations): IL-6 with C-reactive protein (r=0.48), and C-reactive protein with fibrinogen (r=0.41). While no associations were found with concentrations of C-reactive protein or IL-6, urinary albumin excretion was correlated with erythrocyte sedimentation rate (r=0.47) and with fibrinogen concentration (r=0.39; p<0.05). In patients with type 2 diabetes mellitus, urinary albumin excretion was not associated with concentrations of IL-6 or C reactive protein. These results suggest an IL-6-independent mechanism for increased fibrinogen levels and erythrocyte sedimentation rate in type 2 diabetic patients with increased urinary albumin excretion. PMID- 11855793 TI - Efficacy of ACE inhibitors and ATII receptor blockers in patients with microalbuminuria: a prospective study. AB - We compared the efficacy of treatment protocols with an angiotensin converting enzyme (ACE) inhibitor alone (enalapril, 5 mg) or angiotensin II (ATII) receptor blocker (losartan, 50 mg) or both enalapril plus losartan in patients with microalbuminuria in a prospective, randomized clinical trial. Normotensive type 2 diabetic patients with microalbuminuria documented by at least 3 consecutive urinary albumin excretion analyses were recruited for the study. Patients were grouped randomly into one of the protocols which consisted of treatment with 5 mg enalapril daily (group 1; n=12), 50 mg losartan daily (group 2; n=12) or both drugs (group 3; n=10). They were reevaluated with regard to HbA1c levels, lipid profiles, blood pressure and urinary albumin excretion rates (UAER) at 3-month intervals for 12 months. Mean age, duration of diabetes, body mass index, plasma lipid profiles and blood pressure levels were similar at the initial visit. In group 1, UAER returned to normal levels in 10 patients. Normalization of UAER occurred in 8 and 7 patients in groups 2 and 3, respectively. Percentage of reduction in UAERs at the end of 12 months were 58%, 59% and 60% (p=0.0001; p=0.0002; p=0.0003, respectively). The amount of reduction in UAER did not differ significantly among the three groups (p=0.346). ACE inhibitors and angiotensin II receptor blockers have similar efficacy in treating diabetic microalbuminuria, and the combination of the two drugs does not add any further benefit. PMID- 11855794 TI - Effect of glycation of high density lipoproteins on their physicochemical properties and on paraoxonase activity. AB - We investigated the effect of incubation of high density lipoprotein (HDL) under hyperglycaemic conditions on lipid composition, physicochemical properties and activity of paraoxonase (PON), a calcium-dependent enzyme associated with HDL that contributes to the antiatherogenicity of this lipoprotein. HDL incubated for three days with various glucose concentrations (0-100 mM) had significant increases in thiobarbituric acid-reactive substances (TBARS) and conjugated dienes with respect to control HDL. These results suggest that lipid peroxidation accompanies HDL glycation in vitro. The susceptibility to lipid peroxidation was higher in HDL isolated from subjects with low HDL-paraxonase activity with respect to subjects with higher HDL-PON activity. The lipid compositional changes were associated with modifications of apoprotein conformation as shown by the red shifted position of the maximum emission of tryptophan in treated HDL. The decrease in the Gp (generalized polarization) value and the red-shifted position of the maximum emission of Laurdan incorporated in treated HDL demonstrate modifications of order and polarity with respect to control HDL. The negative correlation established between the Gp value and TBARS demonstrates that the modifications in molecular order are likely related to the increase in lipid peroxidation products. The activity of paraoxonase was significantly decreased in HDL incubated at 37 degrees C; a greater decrease occurred in the presence of 50 mM and 100 mM glucose. This study demonstrates modifications of lipid composition, apoprotein conformation and physicochemical properties of HDL incubated in the presence of glucose. These modifications affect the activity of HDL-associated paraoxonase. The physicochemical properties of lipoproteins play a regulatory role in lipoprotein function. The modification of order and polarity of glycated HDL and the alterations in paraoxonase activity could potentially contribute to the accelerated atherosclerosis in diabetic patients. PMID- 11855795 TI - Effect of extracellular pH on insulin secretion and glucose metabolism in neonatal and adult rat pancreatic islets. AB - Changes in extracellular pH are known to affect glucose-stimulated insulin secretion. In the present study, glucose metabolism in pancreatic islets cultured at different pHs was investigated. Also, for islet transplantation purposes, insulin secretion and glucose metabolism were compared in neonatal and adult islets at different pHs to determine which islet preparation is more tolerant to acidity and alkalinity. The results revealed a dependency of insulin secretion on the external pH in both neonatal and adult islets. Reduction of insulin secretion was observed at both the acidic and alkaline sides of pH 7.3. Glucose stimulated increases of insulin secretion in all cases. Similar results were obtained for ATP and pyruvate contents. Intracellular insulin increased with the increase of pH value. In contrast, calcium content decreased with the increase of pH. The results demonstrate that neonatal islets are more acid tolerant than adult islets. Both basal and glucose-stimulated insulin secretions, as well as other parameters of neonatal islets were significantly higher than those of adult islets in response to low pH. The differences under alkaline conditions were not significant but give an indication that neonatal islets are more tolerant to alkalinity than are adult islets. PMID- 11855796 TI - Diabetes-induced elevations in retinal oxidative stress, protein kinase C and nitric oxide are interrelated. AB - Hyperglycemia results in various retinal metabolic abnormalities that can contribute to the development of retinopathy, but it has been difficult to recognize which abnormalities are critical. In this study, the possible interrelationship between hyperglycemia-stimulated oxidative stress, protein kinase C (PKC), and nitric oxide (NO) was investigated by examining the effects of inhibitors of oxidative stress, PKC and NO on glucose-induced retinal oxidative stress, PKC activity and NO levels concentrations, both under in vitro conditions in retinal endothelial cells and isolated retina, and in vivo in the retina from diabetic rats. Bovine retinal endothelial cells were incubated in 5 or 30 mM glucose for 3 days in the presence or absence of inhibitors of oxidative stress (N-acetyl cysteine), PKC (LY333531), or NO (L-NAME). Incubation of retinal endothelial cells in 30 mM glucose resulted in an approximately 2-fold elevation in retinal TBARS, PKC and NO. Addition of N-acetyl cysteine, LY333531, or L-NAME significantly inhibited glucose-induced elevation in oxidative stress, NO and PKC. Similar results were obtained when intact retinas from normal rats were incubated with 30 mM glucose for 6 hours. In diabetic rats, elevations in retinal TBARS, PKC and NO were observed at 2 months of diabetes, and administration of N acetyl cysteine, LY333531 or aminoguanidine prevented diabetes-induced elevation in retinal TBARS and NO levels, and PKC activity. Thus, these results suggest that diabetes-induced metabolic abnormalities, originally considered to be independent abnormalities, are apparently interrelated in retina; inhibiting a single retinal abnormality may have multiple beneficial effects to correct retinal dysmetabolism and to inhibit the development of retinopathy. PMID- 11855797 TI - Advanced oxidation protein products in plasma: stability during storage and correlation with other clinical characteristics. AB - Proteins are susceptible to free radical damage. We measured advanced oxidation protein products (AOPP) in the plasma of 56 hospitalised patients. Concentrations of AOPP were expressed as chloramine-T equivalents by measuring absorbance in acidic conditions at 340 nm in the presence of potassium iodide. We also determined erythrocyte sedimentation rate (ESR), circulating urea, creatinine, glucose, uric acid, electrolytes, lipids, total proteins and fractions and fibrinogen. Twenty-four samples were processed both immediately and after 7, 15, 30, 90, 180 and 438 days of storage at both at -20 degrees C and -80 degrees C (aliquots were frozen and thawed only once) to evaluate AOPP stability. The remaining 32 samples were also processed for thiobarbituric-acid-reactive substances (TBARS). Mean AOPP concentration in all 56 patients was 48.3+/-37.2 microM. Mean basal concentration of AOPP in the 24 plasma samples (55.0+/-47.1 microM) showed no significant change at each intermediate determination, yet significantly increased after 438 days of storage both at -80 degrees C (96.6+/ 83.2, p<0.01) and, markedly, at -20 degrees C (171.3+/-94.6, p<0.001). TBARS concentration was 1.59+/-0.65 micromol/l. Multiple regression analysis evidenced that AOPP concentration was positively correlated (multiple r=0.62, p<0.001) with serum urea and triglycerides, but negatively correlated with patient age (indeed, serum albumin and total proteins decreased with increasing age, r=0.3, p<0.05). TBARS concentration was associated with ESR and serum glucose (multiple r=0.73, p<0.001), yet positively with AOPP (r=0.39, simple p<0.05). We conclude that AOPP remain stable during sample storage both at -20 degrees C and -80 degrees C for 6 months. Renal failure and hypertriglyceridemia probably enhance the in vivo process of AOPP formation. Oxidative damage as measured by TBARS may be increased because of exposure to hyperglycemia causing nonenzymatic glycation of plasma proteins. PMID- 11855799 TI - Adding strings to our bow or being here now? PMID- 11855798 TI - Complete mutation scanning of the human SEL 1L gene: a candidate gene for type 1 diabetes. PMID- 11855800 TI - Therapeutic caring: a learning disability experience. AB - This paper is a reflective account of our experiences of giving Therapeutic Touch (TT), for the first time within a learning disability setting, to a client who has profound learning disabilities. Using a case study approach, we share our story of this pathfinder journey of discovery and show how the process of reflection was instrumental in enabling us to gain insight on the unfolding therapeutic and caring relationship which we developed with this client. Issues relating to informed consent were addressed, however these were complex and needed special consideration. As client consent could not be achieved through verbal means we needed to draw on our senses and use intuitive skills together with team involvement. IntroducingTT into a learning disability setting was breaking new ground, and as this case study has shown it does appear to have the potential to enhance therapeutic caring. PMID- 11855801 TI - Complementary therapies in action--education and outcomes. AB - A study investigated the use of aromatherapy hand and foot massage on 11 patients in a rural rehabilitation setting. An education programme for nurses, carers and family members was developed and implemented. Clinical outcomes--pain, anxiety, joint flexibility and skin condition were evaluated using Likert scales completed before, and after, treatment at three time points and open questions to ascertain patients' feelings about the treatments. Each patient had three treatments, making a total of 33 massages. A significant reduction in pain and anxiety was apparent after all three treatments (p=0.05). Changes in skin condition (softer and more resilient) were highly significant (p=0.01). However, there was no significant change in joint flexibility (p<0.05). The main themes emerging from patients'comments were that aromatherapy massage facilitated communication, allowed emotional release and aided relaxation. Nine people undertook the education programme. They indicated that it covered appropriate information and they felt confident to deliver the aromatherapy massages, but the non-nursing participants would have liked more practice before they entered the clinical setting. PMID- 11855802 TI - The midwife as healer. AB - In ancient times the midwife was often also a healer, serving communities and women and passing knowledge and insights from woman to woman, mother to daughter. Much of this knowledge has been 'lost' or 'forgotten' due to the witch hunts of the Middle Ages and the discrediting of women's knowledge as scientific rationality emerged. There is evidence now that midwives are exploring alternative knowledge in their rejection of the medical model of care and in their quest to provide enhanced and holistic women-centred care. The three areas highlighted--utilization of complementary therapies, exploration of spirituality and the examination of the nature of the mother--midwife relationship--represent exciting evidence that the midwife is beginning to re-embrace the role of healer. PMID- 11855803 TI - Traditional Chinese customs and practices for the postnatal care of Chinese mothers. PMID- 11855804 TI - Utilizing the mythical and folkloric power of trees in the modern hospital environment. AB - The Department of Health, under the auspices of the Government of the United Kingdom, has recently been encouraging hospital management boards to plant trees for the benefit of patients and staff. Although there is little scientific evidence to support such an edict, there are thousands of years of mythical and folkloric evidence to support such an initiative. This evidence is briefly reviewed, paying particular attention to the supposed powers often ascribed to oak, rowan, yew, hawthorn, ash and beech trees. Recommendations for arboreal tree planting schemes in hospitals are made based on this evidence. PMID- 11855805 TI - Food, fractions and complementary therapy. PMID- 11855806 TI - Veal L 2001. A comparison of the use of complementary therapies in Australia and Iceland. Complementary therapies in nursing & midwifery 7: 72-77. PMID- 11855807 TI - Doxorubicin and C-13 deoxydoxorubicin effects on ryanodine receptor gene expression. AB - Chronic anthracycline administration to rabbits causes impairment of cardiac contractility and decreased gene expression of the calcium-induced calcium release channel of sarcoplasmic reticulum (SR), the ryanodine receptor (RYR2). The C-13 hydroxy metabolite (doxorubicinol), formed in the heart, has been hypothesized to contribute to anthracycline cardiotoxicity. C-13 deoxydoxorubicin is an analog unable to form the C-13 hydroxy metabolite. Therefore, doxorubicin, C-13 deoxydoxorubicin, or saline was administered to rabbits (1 mg/kg iv twice weekly for 8 weeks). Left ventricular fractional shortening (LVFS) was decreased by chronic treatment with doxorubicin (28 +/- 2%; P < 0.05), but not C-13 deoxydoxorubicin (33 +/- 2%) compared to age-matched pair-fed controls. Doxorubicin, but not C-13 deoxydoxorubicin, caused a significant reduction (P < 0.02) in the ratio of RYR2/Ca-Mg ATPase (SERCA2) mRNA levels (0.57 +/- 0.1 vs 1.22 +/- 0.2, respectively) in the left ventricle. This suggests that doxorubicinol may contribute to the downregulation of cardiac RYR2 expression in chronic doxorubicin cardiotoxicity. PMID- 11855808 TI - Insulin-stimulated fatty acid synthase gene expression does not require increased sterol response element binding protein 1 transcription in primary adipocytes. AB - Sterol response element binding protein 1c (SREBP-1c) is a transcription factor that has been implicated in the regulation of expression of key lipogenic genes in hepatocytes, including fatty acid synthase (FAS) and glucokinase. In hepatocytes, insulin stimulates a rapid increase in transcription of SREBP-1c and the appearance of the SREBP-1c protein in the nucleus. SREBP-1 has also been proposed to play an important role in the induction of expression of lipogenic enzymes in adipose tissue in vivo in response to nutritional status. In this paper we have investigated the regulation of the SREBP-1 and FAS genes in adipocytes and find that while an overexpressed constitutively active SREBP-1 mutant is capable of substantially stimulating the FAS promoter, insulin appears to stimulate FAS gene expression in primary adipocytes in the absence of any apparent effect on SREBP-1 transcription. Taken together, our data suggest that insulin does not stimulate FAS gene expression through increasing SREBP-1c transcription in adipose cells. PMID- 11855809 TI - Protein kinase D is a downstream target of protein kinase Ctheta. AB - Protein kinase D (PKD/PKCmu immunoprecipitated from either COS-7 cells or Jurkat T lymphocytes transiently transfected with a constitutively active mutant of PKCtheta AE (PKCthetaAE) exhibited a marked increase in basal activity. In contrast, coexpression of constitutively active mutant of PKCzeta does not induce PKD activation in both types of cells. PKCthetaAE does not induce kinase activity in immunocomplexes of PKD kinase-deficient mutants PKDK618N or PKDD733A. PKD activation in response to PKCthetaAE signaling was completely prevented by treatment with the protein kinase C (PKC) inhibitors, GF I or Ro 31-8220, or by mutation of Ser-744 and Ser-748 to Ala in the kinase activation loop of PKD. Our results show that PKD is a downstream target of the theta isoform of PKC in both COS-7 cells and lymphocytes. The regulation of PKD by PKCtheta reveals a new pathway in the signaling network existing between multiple members of the PKC superfamily and PKD. PMID- 11855810 TI - Heat shock inhibits activation of NF-kappaB in the absence of heat shock factor 1. AB - The heat shock response is known to inhibit NF-kappaB activation and NF-kappaB dependent gene expression. Herein we determined if cells lacking heat shock factor-1 (HSF-1), the major transcription factor regulating heat shock protein gene expression, have an altered ability to modulate NF-kappaB activation. Embryonic fibroblasts from HSF-1-null mutant mice (HSF-1-/- cells) had a drastically reduced ability to express heat shock protein-70 in response to heat shock, compared to embryonic fibroblasts from wild-type mice (HSF+/+ cells). There was no difference, however, in the ability of heat shock to inhibit TNFalpha-mediated NF-kappaB activation, IkappaBalpha degradation, IkappaB kinase activation, and macrophage chemotactic protein-1 expression in the HSF-1-/- cells compared to the HSF-1+/+ cells. These data demonstrate that heat shock inhibits activation of the NF-kappaB/IkappaBalpha pathway and NF-kappaB-dependent gene expression in the absence of an intact heat shock response. PMID- 11855811 TI - Dominant negative alpha-subunit of FTase inhibits effects of insulin and IGF-I in MCF-7 cells. AB - We recently designed a dominant negative (DN) farnesyltransferase (FTase)/geranyl gerahyltransferase I (GGTase I) alpha-subunit that when expressed in vascular smooth muscle cells decreased insulin-stimulated phosphorylation of FTase, FTase activity, amounts of farnesylated p21Ras, DNA synthesis, and cell migration. Currently, we explored the inhibitory effects of DN FTase/GGTase I alpha-subunit in MCF-7 cells on IGF-1- and insulin-stimulated DNA synthesis and cell proliferation. Expression of the DN FTase/GGTase I alpha-subunit completely blocked IGF-1- and insulin-stimulated BrdU incorporation and cell count. DN FTase/GGTase I alpha-subunit inhibited insulin-stimulated phosphorylation of FTase/GGTase I alpha-subunit, FTase and GGTase I activity, and prenylation of p21Ras and RhoA. Expression of DN FTase/GGTase I alpha-subunit diminished IGF-1- and insulin-stimulated phosphorylation of ERK (extracellular signal-regulated kinase), but had no effect on IGF-1- and insulin-stimulated phosphorylation of Akt. Taken together, these data suggest that DN FTase/GGTase I alpha-subunit can assuage the mitogenic effects of IGF-1 and insulin on MCF-7 breast cancer cells. PMID- 11855812 TI - MUC17, a novel membrane-tethered mucin. AB - Membrane mucins have several functions in epithelial cells including cytoprotection, extravasation during metastases, maintenance of luminal structure, and signal transduction. In this paper we describe a large membrane mucin expressed in the normal intestine. This novel mucin, designated MUC17, contains an extended, repetitive extracellular glycosylation domain and a carboxyl terminus with two EGF-like domains, a SEA module domain, a transmembrane domain, and a cytoplasmic domain with potential serine and tyrosine phosphorylation sites. RNA blot analysis and in situ hybridization indicates that MUC17 is expressed in select pancreatic and colon cancer cell lines and in intestinal absorptive cells. Radiation hybrid mapping localized MUC17 to chromosome 7q22 where it resides in close proximity with three other membrane mucin genes, MUC3A, MUC3B, and MUC12. Thus, these membrane mucins reside together in a gene cluster, but are expressed in different tissues and are likely to have different functions as well. PMID- 11855813 TI - Occurrence of Cu-ATPase in Dictyostelium: possible role in resistance to copper. AB - Dictyostelium discoideum amoebae showed an uncommon resistance to Cu(2+), as pointed out through cell growth rate (EC(50) = 469 +/- 30 microM) and the neutral red cytotoxicity assay (EC(50) = 334 +/- 45 microM). Although no evidence of Cu inducible metallothionein was found, Cu-dependent ATPase activity was cytochemically detected on pelletted, resin-embedded amoebae. This activity required Cu(2+) in the incubation medium, was sensitive to TPEN, vanadate and temperature, and showed dose-dependent increase after exposure of amoebae to 10 500 microM Cu(2+) for 7 days. Accordingly, immunofluorescence and Western blotting revealed the occurrence of a Cu-inducible, putative homologue of human Menkes (MNK) Cu-P-type ATPase. To verify if Cu-ATPase is involved in copper resistance, amoebae were exposed to low concentrations of Cu(2+) and vanadate followed by the neutral red assay. Exposure to either treatment showed no effect, while a combination caused a dramatic increase of Cu toxicity, possibly depending on Cu-ATPase inhibition. PMID- 11855814 TI - Death effector domain-only polypeptides of caspase-8 and -10 specifically inhibit death receptor-induced cell death. AB - Caspase-8 and -10 are thought to be involved in a signaling pathway leading to death receptor-mediated apoptosis. The prodomains of these caspases are known to form fibrous structures in the perinuclear region when overexpressed, though the meaning of the structures remains unclear. In a previous study we showed that the overexpressed caspase-8 or -10 prodomain (PDCasp8 or PDCasp10) did not induce cell death, and we hypothesized that these prodomains interfere with the receptor mediated cell death signaling pathway. Indeed, in 293, HeLa and Jurkat cells, cell death mediated by agonistic anti-Fas antibody, TRAIL or overexpression of full-length caspase-8 was significantly inhibited by overexpression of PDCasp8 or PDCasp10 which colocalized with the Golgi complex and with overexpressed FADD. However, when about 20 amino acid residues were deleted from either terminus of the caspase-10 prodomain (amino acid residue 1 to 219), the ability to inhibit Fas-mediated cell death was lost. Interestingly, these deletion mutants also lost the ability to make fibrous structures and to bind FADD, suggesting that FADD binding is important for their function, and that PDCasp8 and PDCasp10 act as dominant-negative inhibitors. PMID- 11855816 TI - A discrete domain of the human TrkB receptor defines the binding sites for BDNF and NT-4. AB - TrkB is a member of the Trk family of tyrosine kinase receptors. In vivo, the extracellular region of TrkB is known to bind, with high affinity, the neurotrophin protein brain-derived neurotrophic factor (BDNF) and neurotrophin-4 (NT-4). We describe the expression and purification of the second Ig-like domain of human TrkB (TrkBIg(2)) and show, using surface plasmon resonance, that this domain is sufficient to bind BDNF and NT-4 with subnanomolar affinity. BDNF and NT-4 may have therapeutic implications for a variety of neurodegenerative diseases. The specificity of binding of the neurotrophins to their receptor TrkB is therefore of interest. We examine the specificity of TrkBIg(2) for all the neurotrophins, and use our molecular model of the BDNF-TrkBIg(2) complex to examine the residues involved in binding. It is hoped that the understanding of specific interactions will allow design of small molecule neurotrophin mimetics. PMID- 11855817 TI - Nuclear translocation of mouse polycomb m33 protein in regenerating liver. AB - Immunoblots probed with an antibody to M33 protein, a homolog of Drosophila Polycomb, revealed that most M33 in adult mouse liver had a higher electrophoretic mobility than that in F9 embryonal carcinoma cells. High-mobility 60-kDa M33 localized in the cytoplasmic fraction of liver homogenates, and two less abundant 66- and 70-kDa species were detected in the nuclear fraction. Immunocytochemistry of freeze-substituted tissues showed a punctate pattern of immunofluorescence in the cytoplasm of hepatic parenchymal cells. Nuclear M33 isoforms treated with alkaline phosphatase had increased mobilities corresponding to cytoplasmic M33. In partially hepatectomized mice, nuclear M33 isoforms appeared after 48 h, near the time of maximum DNA synthesis as measured by bromodeoxyuridine incorporation. By 60 h, most M33 was in the form of these low mobility species, and the pattern of immunofluorescence suggested the existence of chromatin-bound and free states of the protein in the nucleus. Thereafter, high-mobility 60-kDa M33 reappeared. The data are consistent with a phosphorylation-associated translocation mechanism that is a cell cycle dependent. PMID- 11855815 TI - Possible implication of Golgi-nucleating function for the centrosome. AB - The Golgi apparatus breaks down at mitosis, resulting in the dispersal of Golgi resident proteins. In NRK cells, however, subsets of both TGN38 and golgin-97, but not ManII and GM130, remained associated with the centrosome throughout the cell cycle. This centrosome association of TGN38 and golgin-97 was not disrupted by treatment with brefeldin A, additional inducers of retrograde trafficking and inhibitors of either kinases or protein phosphatases. Anchoring of the Golgi apparatus within the juxtanuclear region depends on microtubules; the association of TGN38 and golgin-97 subsets with the centrosome, however, was insensitive to nocodazole treatment. Drugs such as PDMP, which block Golgi dispersal both by nocodazole, despite microtubule depolymerization, and by inducers of retrograde trafficking, strengthened the microtubule-nucleating activity of the centrosome. These observations cumulatively suggest the centrosome is implicated in nucleation of the Golgi apparatus through interactions with Golgi-resident proteins, such as TGN38 and golgin-97. PMID- 11855818 TI - Regulation of Trypanosoma cruzi invasion of nonphagocytic cells by the endocytically active GTPases dynamin, Rab5, and Rab7. AB - During invasion of nonphagocytic cells by Trypanosoma cruzi (T. cruzi), host cell lysosomes are recruited to the plasma membrane attachment site followed by lysosomal enzyme secretion. The membrane trafficking events involved in invasion have not been delineated. We demonstrate here that T. cruzi invasion of nonphagocytic cells was completely abolished by overexpression of a dominant negative mutant of dynamin. Likewise, overexpression of a dominant negative mutant of Rab5, the rate-limiting GTPase for endocytosis, resulted in reduced infection rates compared with cells expressing Rab5 wild-type. Moreover, cells expressing the activated mutant of Rab5 experienced higher infection rates. A similar pattern was also observed when Rab7-transfected cells were examined. Confocal microscopy experiments showed that parasites colocalized with green fluorescent protein-Rab5-positive early endosomes after 5 min of invasion. These data clearly indicate that newly forming T. cruzi phagosomes first interact with an early endosomal compartment and subsequently with other late component markers before lysosomal interaction occurs. PMID- 11855819 TI - Characterization of adiposity and metabolism in Lmna-deficient mice. AB - Dunnigan's Familial Partial Lipodystrophy (FPLD) is an autosomal dominant disease characterized by regional fat loss and insulin resistance. FPLD is caused by mutations in the LMNA gene, which encodes intermediate filaments of the nuclear lamina. Different LMNA mutations cause Emery-Dreifuss muscular dystrophy and/or a dilated cardiomyopathy. It is not known how LMNA mutations cause any of the disease phenotypes. Here we measure physical and metabolic characteristics of Lmna-/- and +/- mice to determine their usefulness as models for FPLD. Lmna-/- mice, which die prematurely of muscular dystrophy, have little fat, but do not show the insulin resistance characteristic of FPLD. Lmna+/- mice, despite treatment with a high fat diet, do not have decreased fat stores or metabolic features of FPLD. We also show, in mice, that Lmna transcripts are expressed at high levels in muscle and adipose tissue, but do not vary by body region or sex. In conclusion, Lmna+/- and -/- mice do not mimic Dunnigan's FPLD, and differential expression of lamins A and C does not appear to contribute to sex- or tissue-specific LMNA phenotypes. PMID- 11855820 TI - Use of gDNA as internal standard for gene expression in staphylococci in vitro and in vivo. AB - An internal RNA standard proved less suitable in bacterial gene expression experiments. We therefore developed a method for simultaneous RNA and gDNA (genomic DNA) isolation from in vitro and in vivo samples containing staphylococci and combined it with quantitative PCR. The reliability of gDNA for bacterial quantification and for standardisation in gene expression experiments was evaluated. Quantitative PCR proves equivalent to quantitative culture for in vitro samples, and superior for in vivo samples. In gene expression experiments, gDNA permits a good standardisation for the initial amount of bacteria. The average interassay variability of the in vitro expression is 20.1%. The in vivo intersample variability was 73.3%. This higher variability can be attributed to the biological variation of gene expression in vivo. This method permits exact quantification of the number of bacteria and the expression of genes in staphylococci in vivo (e.g., in biofilms, evolution in time) and in vitro. PMID- 11855821 TI - Structural domains of vault proteins: a role for the coiled coil domain in vault assembly. AB - Vaults consist of multiple copies of three proteins (MVP, VPARP, and TEP1) and several untranslated RNAs. The function of vaults is unknown but the typical and evolutionary conserved structure indicates a role in intracellular transport. Although all vault components have been identified and characterized, not much is known about vault protein assembly. In this study we identified and analyzed structural domains involved in vault assembly with emphasis on protein-protein interactions. Using a yeast two-hybrid system, we demonstrate within MVP an intramolecular binding site and show that MVP molecules interact with each other via their coiled coil domain. We show that purified MVP is able to bind calcium, most likely at calcium-binding EF-hands. No interactions could be detected between TEP1 and other vault proteins. However, the N-terminal half of MVP binds to a specific domain in the C-terminus of VPARP. Furthermore, VPARP contains amino acid stretches mediating intramolecular binding. PMID- 11855822 TI - Inhibition of the ubiquitin-proteasome pathway induces differential heat-shock protein response in cardiomyocytes and renders early cardiac protection. AB - The effects of proteasome inhibition (PI) on heat-shock protein (HSP) expression in cardiomyocytes were investigated. Neonatal rat cardiomyocytes were incubated with MG132 (0.1-10 microM) for 1 h. Induction of various HSPs was determined by real-time PCR and Western blotting. PI induced a 2- to 3-fold increase in HSP27, HSP60, and HSP90, and a 18-fold increase in HSP70 mRNA expression, whereas HSP40 levels were unaffected. Western blotting revealed increased protein expression for HSP70 after PI. Similar results were obtained with MG262. HSP induction correlated with enhanced survival of neonatal cardiomyocytes after sublethal heat stress in XTT testing. In papillary muscles, pretreatment with MG132 (10 microM, 90 min) was associated with enhanced recovery of the contractile parameters after a 40-min hypoxia. In these proof-of-principle experiments, we show that PI induces differential heat-shock response in cardiomyocytes, accompanied by enhanced cell survival and functional recovery after various forms of stress. PMID- 11855823 TI - Ebselen has dehydroascorbate reductase and thioltransferase-like activities. AB - Ebselen (2-phenyl-1,2-benzisoselenazol-3(2H)-one), a seleno-organic compound, has been reported to mimic glutathione peroxidase (GPX). Since bovine erythrocyte GPX showed dehydroascorbic acid (DHA) reductase and thioltransferase (TTase) activities, ebselen was also examined for DHA reductase and TTase-like activities. Evidence is reported that, in the presence of GSH, ebselen catalyzed the in vitro reduction of DHA to L-ascorbic acid in a dose-dependent manner. Using S-sulfocysteine and GSH as co-substrates, ebselen catalyzed the in vitro formation of glutathione disulfide in a dose-dependent manner, thereby acting as a TTase mimic. 1-Chloro-2,4-dinitrobezene (CDNB), a co-substrate with GSH for glutathione S-transferase, was used to measure rates of adduct formation with ebselen pretreated with GSH and compared with GSH alone. The reaction rate was proportional to ebselen, and ebselen was about 250 times more reactive than GSH on an equimolar basis. The DHA reductase and TTase-like activities, in addition to the powerful nucleophilic reactivity of ebselen selenol, may contribute to ebselen's significant anti-inflammatory and anti-oxidative properties in vivo. PMID- 11855824 TI - Soluble Flt-1 (soluble VEGFR-1), a potent natural antiangiogenic molecule in mammals, is phylogenetically conserved in avians. AB - The flt-1 gene encodes for both the full-length receptor Flt-1 (VEGFR-1) and a soluble form designated sFlt-1. sFlt-1 carries the VEGF-binding domain of Flt-1 as well as a 31-amino-acid stretch derived from an intron and tightly binds VEGF, suppressing its angiogenic activity. The flt-1 gene has so far been identified only in mammals and is highly expressed in placenta as well as in vascular endothelial cells. In placenta, sFlt-1 is abundant in the trophoblast layer during pregnancy, suggesting that it is a negative regulator to excess angiogenesis and vascular permeability at the feto-maternal border in mammals. However, we show here for the first time that the flt-1 gene exists and is highly conserved in chickens. Surprisingly, the chicken flt-1 gene also encodes for sFlt 1 in addition to the full-length receptor. Similar to the mammalian sFlt-1, chicken sFlt-1 carries the VEGF-binding domain and a 31-amino-acid carboxyl region derived from an intron, which was significantly homologous to that in mammals. Chicken sFlt-1 is expressed early in embryogenesis. These findings strongly suggest that the natural antiangiogenic molecule sFlt-1 is widely conserved in vertebrates and regulates the angiogenic process. PMID- 11855825 TI - Regulation of cap-dependent translation by insulin-like growth factor-1 in neuronal cells. AB - Insulin-like growth factor-1 (IGF-1) both promotes survival and activates protein synthesis in neurons. In the present paper, we investigate the effect of IGF-1 treatment on cap-dependent translation in primary cultured neuronal cells. IGF-1 treatment increased the phosphorylation of eukaryotic initiation factor (eIF)-4E binding protein 1 (4E-BP1), exclusively at Thr-36 and Thr-45 residues, and eIF-4G phosphorylation at Ser-1108. In contrast, a significant eIF-4E dephosphorylation was found. In parallel, increased eIF-4E/4G assembly and protein synthesis activation in response to IGF-1 treatment were observed. The phosphatidylinositol 3-kinase (PI3-K) inhibitor wortmannin and the mammalian target of rapamycin (mTOR) inhibitor rapamycin, but not the mitogen-activated protein kinase (MAPK) activating kinase (MEK) inhibitor PD98059, reversed the IGF-1-induced effects observed on eIF-4E/4G assembly and phosphorylation status of 4E-BP1, eIF-4E, and eIF-4G. Therefore, our findings show that the IGF-1-induced regulation of cap dependent translation is largely dependent on the PI-3K and mTOR pathway in neuronal cells. PMID- 11855826 TI - Cloning and characterization of the human BAZF gene, a homologue of the BCL6 oncogene. AB - The BAZF gene has recently been identified as a novel homologue of the BCL6 oncogene. Here we cloned the human BAZF gene using murine BAZF as a probe. The predicted amino acid sequence was 91% identical to that of murine BAZF. The BTB/POZ and zinc finger domains were almost completely conserved between human and murine BAZF. Fluorescence in situ hybridization analysis revealed that the human BAZF gene is located on chromosome 17p13.1. Although expression of human BAZF mRNA was ubiquitously detected in human tissues, abundant expression was detected in heart and placenta. BAZF mRNA was expressed in some immature B cell lines and erythroleukemia cell lines. The expression in a human erythroleukemia cell line, HEL cells, was upregulated during megakaryocytic differentiation induced by 12-O-tetradecanoyl-phorbol-13-acetate. These expression patterns of BAZF mRNA suggest that BAZF may regulate differentiation in stages or lineages that are different from those regulated by BCL6. PMID- 11855827 TI - Phosphorylation of the N-terminal and C-terminal CD3-epsilon-ITAM tyrosines is differentially regulated in T cells. AB - Tyrosine phosphorylation of immunoreceptor tyrosine-based activation motifs (ITAMs) within CD3 chains is crucial for the recruitment of protein tyrosine kinases and effector molecules into the T cell receptor. Thus, phenylalanine substitution at the N-terminal tyrosine residue of the CD3-epsilon-ITAM abolished signal transduction functions of this ITAM, including phosphorylation at the C terminal ITAM tyrosine, and its association with ZAP-70. In contrast, mutation at the C-terminal tyrosine of CD3-epsilon-ITAM did not prevent phosphorylation at the N-terminal tyrosine, nor its association with Lck, or p85 PI 3-K regulatory subunit. In contrast to the ZAP-70/diphosphorylated CD8-epsilon-ITAM interaction, the Lck/monophosphorylated CD8-epsilon-ITAM interaction was sensitive to octylglucoside, an agent that disrupts Lck interaction with membrane rafts. Therefore, association of Lck with membrane rafts seems to be essential for stabilization of Lck/CD3-epsilon protein-protein interactions. Overall, the data suggest that the sequential and coordinated phosphorylation of CD3-epsilon-ITAM tyrosines provides to CD3-epsilon the potential to interact with multiple downstream effectors and signaling pathways. PMID- 11855828 TI - Hes-1, a known transcriptional repressor, acts as a transcriptional activator for the human acid alpha-glucosidase gene in human fibroblast cells. AB - Hes-1, the mammalian homologue 1 of Drosophila hairy and Enhancer of split proteins, belongs to a family of basic helix-loop-helix proteins that are essential to neurogenesis, myogenesis, hematopoiesis, and sex determination. Hes 1 is a transcriptional repressor for a number of known genes including the human acid alpha-glucosidase (GAA) gene as we have previously shown in Hep G2 cells. The human GAA gene encodes the enzyme for glycogen breakdown in lysosomes, deficiency of which results in Glycogen Storage Disease type II (Pompe syndrome). Using constructs containing the DNA element that demonstrates repressive activity in Hep G2 cells and conditions in which the same transcription factors, Hes-1 and YY1, bind, we have shown that this element functions as an enhancer in human fibroblasts. Site-directed mutagenesis and overexpression of Hes-1 showed that Hes-1 functions as a transcriptional activator. The dual function of Hes-1 we have found is likely to contribute to the subtle tissue-specific control of this housekeeping gene. PMID- 11855829 TI - Differential role of MAP kinases in stimulation of hepatocyte growth by EGF and G protein-coupled receptor agonists. AB - Several agonists acting on G-protein-coupled receptors (GPCR) enhance the mitogenic effect of EGF in rat hepatocytes. Previous studies have shown that mitogen-activated protein (MAP) kinases are involved in the mitogenic effect of EGF. In the present study on cultured rat hepatocytes we show that although the comitogenic GPCR agonists prostaglandin F(2alpha), vasopressin, angiotensin II, and norepinephrine all activated ERK, blocking of the ERK pathway with the MEK inhibitor PD 98059 did not abolish their comitogenic effects. These GPCR agonists also activated p38, but the p38 blocker SB 203580 did not reduce the comitogenic effects. The mitogenic effect of EGF was inhibited completely by PD 98059 and partially by SB 203580. These results suggest that, in contrast to the mitogenic effect of EGF, the comitogenic effect of a group of GPCR agonists is independent of ERK and p38 in these cells. PMID- 11855830 TI - Substrate specificity of penicillin acylase from Streptomyces lavendulae. AB - The kinetic parameters of several substrates of penicillin acylase from Streptomyces lavendulae have been determined. The enzyme hydrolyses phenoxymethyl penicillin (penicillin V) and other penicillins with aliphatic acyl-chains such as penicillin F, dihydroF, and K. The best substrate was penicillin K (octanoyl penicillin) with a k(cat)/K(m) of 165.3 mM(-1) s(-1). The enzyme hydrolyses also chromogenic substrates as NIPOAB (2-nitro-5-phenoxyacetamido benzoic acid), NIHAB (2-nitro-5-hexanoylamido benzoic acid) or NIOAB (2-nitro-5-octanoylamido benzoic acid), however failed to hydrolyse phenylacetil penicillin (penicillin G) or NIPAB (2-nitro-5-phenylacetamido benzoic acid) and penicillins with polar substituents in the acyl moiety. These results suggest that the structure of the acyl moiety of the substrate is more determinant than the amino moiety for enzyme specificity. The enzyme was inhibited by several organic acids and the extent of inhibition changed with the hydrophobicity of the acid. The best inhibitor was octanoic acid with a K(i) of 0.8 mM. All the results, taking together, point to an active site highly hydrophobic for this penicillin acylase from Streptomyces lavendulae. PMID- 11855831 TI - The ATP binding cassette transporter A1 contributes to the secretion of interleukin 1beta from macrophages but not from monocytes. AB - Deficiency of ABCA1 causes high density lipoprotein deficiency and macrophage foam cell formation in Tangier disease. ABCA1 was also postulated to mediate the secretion of IL-1beta from monocytes and macrophages. We investigated the contribution of ABCA1 to IL-1beta secretion from human monocytes and macrophages of normal donors and Tangier disease patients. Neither an anti-ABCA1 antisense oligonucleotide nor ABCA1 deficiency interfered with LPS-induced secretion of IL 1beta from full blood or freshly isolated monocytes. By contrast, anti-ABCA1 antisense oligonucleotides decreased the LPS-induced secretion of IL-beta from macrophages by 30-50%. The secretion of the precursor pro-IL-1beta and TNFalpha was not inhibited. Compared to normal macrophages, LPS-stimulated Tangier disease macrophages secreted less IL-1beta relative to TNFalpha. Also the spontaneous secretion of IL-1beta by Tangier macrophages was lower than by control cells. We conclude that IL-1beta is secreted from monocytes by an ABCA1-independent pathway and from macrophages by ABCA1-dependent and -independent pathways. PMID- 11855832 TI - Catalytic antibody therapy against the insecticide carbaryl. AB - Catalytic antibodies have been studied widely, but little is known about their applicability as therapeutic reagents in vivo. Here we report that carbaryl, a widely used broad-spectrum carbamate insecticide that is toxic to animals and humans, is hydrolyzed by polyclonal catalytic antibodies induced in vivo by a phosphate immunogen. To test the efficacy of the in vivo-induced polyclonal antibodies, we immunized mice with the phosphate immunogen and assayed their sensitivity to carbaryl by determining the ED(50) value, the dose that produces lowest-grade tremors in 50% of animals. We found that the ED(50) for immunized mice was 43% higher than that for nonimmunized mice and that this increase in ED(50) probably resulted from the hydrolysis of carbaryl by the catalytic antibodies in vivo. Our results suggest that polyclonal catalytic antibodies can be used as therapeutic reagents in vivo. PMID- 11855833 TI - P2X(1) receptor subunit contribution to gating revealed by a dominant negative PKC mutant. AB - The family of ATP-gated P2X receptor channels have a conserved protein kinase C site in the N-terminal intracellular domain. This site was disrupted in human P2X(1) receptors by the mutation T18A. T18A mutants were expressed at normal levels in Xenopus oocytes; however, the peak current amplitude was reduced by >99% and showed approximately 10 fold faster desensitisation in response to ATP than wild type (WT) receptors showed. P2X receptor subunits form functional trimeric channels. Co-expression of T18A and WT receptors (90:10 ratio) produced heteromeric T18A/WT channels with the rapid T18A time-course and an approximately 90-fold increase in peak current amplitude compared to T18A. Similarly, T18A dominated the desensitisation phenotype of heteromeric channels composed of T18A and slowly desensitising K68A mutants. These results suggest that phosphorylation of P2X(1) receptors has a dramatic effect on the time-course of the response and may provide a mechanism for regulating channel function. PMID- 11855834 TI - Role of the Multidrug Resistance Protein 1 in protection from heavy metal oxyanions: investigations in vitro and in MRP1-deficient mice. AB - The Multidrug Resistance Protein 1 (MRP1) is a membrane pump that mediates the efflux of a wide variety of xenobiotics, including arsenical and antimonial compounds, as demonstrated by the study of MRP1-transfected cell lines. We have previously shown that mrp1(-/-) cells are hypersensitive to sodium arsenite, sodium arsenate, and antimony potassium tartrate. We now report that the retroviral vector-mediated overexpression of MRP1 and of the two subunits of gamma-GCS (heavy and light) resulted in higher intracellular glutathione levels and in a greater level of resistance to sodium arsenite and antimony potassium tartrate, compared to the overexpression of MRP1 and gamma-GCS heavy alone. These observations further demonstrate that glutathione is an important component of MRP1-mediated cellular resistance to arsenite and antimony. However, the constitutive expression of MRP1 did not protect mice from the lethality of sodium arsenite and antimony potassium tartrate nor reduced the tissue accumulation of arsenic in mice injected i.p. with sodium arsenite. It is conceivable that, in vivo, other pump(s) effectively vicariate for MRP1-mediated transport of heavy metal oxyanions. PMID- 11855835 TI - Restricted expression of calcium-binding protein S100A5 in human kidney. AB - Reverse transcription--polymerase chain reaction (RT-PCR) identified the expression of calcium-binding protein S100A5 in the noncancerous parts of resected samples from renal cell carcinoma (RCC) patients (n = 7) but not in the carcinoma lesions. Rabbit anti-S100A5 antibody immunohistochemically detected the antigen in the thick ascending limb of Henle, distal convoluted tubule, and collecting duct system. No apparent immunopositivity was observed in the glomerulus, proximal tubules, interstitial cells, or RCC cells. Thus, it was suggested that S100A5 protein plays an inherent functional role to the post-thick ascending limb of Henle portion in the nephron. Further, the carcinomas tested were originated probably not in the S100A5-positive distal epithelium but in the negative epithelium of proximal tubules. Then, total RNA was extracted by phenol/chloroform from 1 ml urine of healthy volunteers, and S100A5 was amplified by RT-PCR from all samples (n = 12), indicating that the transcript of S100A5 is detectable even in the cells released into urine. PMID- 11855836 TI - Phosphorylation of Fanconi anemia protein, FANCA, is regulated by Akt kinase. AB - Phosphorylation of the Fanconi anemia complementation group A (FANCA) protein is thought to be important for the function of the FA pathway. However, the kinase for FANCA (so-called FANCA-PK) remains to be identified. FANCA has a consensus sequence for Akt kinase near serine 1149 (Ser1149), suggesting that Akt can phosphorylate FANCA. We performed in vitro kinase assays using as substrate either a GST-fusion wild-type (WT) FANCA fragment or a GST-fusion FANCA fragment containing a mutation from serine to alanine at 1149 (FANCA-S1149A). These experiments confirmed that FANCA is phosphorylated at Ser 1149, in vitro. However, (32)P-orthophosphate labeling experiments revealed that FANCA-S1149A was more efficiently phosphorylated than WT-FANCA. Furthermore, phosphorylation of wild-type FANCA was blocked by coexpression of a constitutively active (CA)-Akt and enhanced by a dominant-negative (DN) Akt. Our results suggest that Akt is a negative regulator of FANCA phosphorylation. PMID- 11855837 TI - Characterization of a Kunitz trypsin inhibitor with one disulfide bridge purified from Swartzia pickellii. AB - Swartzia pickellii is a Leguminosae that belongs to the Caesalpinioideae sub family the Swartzia pickellii Trypsin Inhibitor (SWTI), a serine proteinase inhibitor was isolated from its seeds. SWTI is a single polypeptide chain protein and it's structure has 174 amino acid residues, it homologous to other Kunitz plant inhibitors, however shows some major differences: it contains only one disulfide bridge, instead two which are usually found in plant Kunitz inhibitors, and the SWTI reactive site does not contain the usual Arg or Lys residues at the putative reactive site (position 65). A glycosylation site was detected at Asn38 with 1188 kDa carbohydrate portion. The primary structure micro heterogeneity was found combining the sequence determination and mass spectrometry. Three forms of SWTI were actually defined: two glycosylated forms a 20,204 kDa (Arg 165) and 20,185 kDa (His 165) and one deglycosylated form 19,016 kDa (Arg 165), all of them contain a Met residue at position 130. PMID- 11855838 TI - Synthesis, 3-D structure, and pharmacology of a reticulated chimeric peptide derived from maurotoxin and Tsk scorpion toxins. AB - Maurotoxin (MTX) is a 34-mer scorpion toxin cross-linked by four disulfide bridges that acts on both Ca(2+)-activated (SK) and voltage-gated (Kv) K(+) channels. A 38-mer chimera of MTX, Tsk-MTX, has been synthesized by the solid phase method. It encompasses residues from 1 to 6 of Tsk at N-terminal, and residues from 3 to 34 of MTX at C-terminal. As established by enzyme cleavage, Tsk-MTX displays half-cystine pairings of the type C1-C5, C2-C6, C3-C7 and C4-C8 which, contrary to MTX, correspond to a disulfide bridge pattern common to known scorpion toxins. The 3-D structure of Tsk-MTX, solved by (1)H NMR, demonstrates that it adopts the alpha/beta scaffold of scorpion toxins. In vivo, Tsk-MTX is lethal by intracerebroventricular injection in mice (LD(50) value of 0.2 microg/mouse). In vitro, Tsk-MTX is as potent as MTX, or Tsk, to interact with apamin-sensitive SK channels of rat brain synaptosomes (IC(50) value of 2.5 nM). It also blocks voltage-gated K(+) channels expressed in Xenopus oocytes, but is inactive on rat Kv1.3 contrary to MTX. PMID- 11855839 TI - Islet-neogenesis-associated protein enhances neurite outgrowth from DRG neurons. AB - Islet-neogenesis-associated protein, INGAP, is a 175-amino-acid pancreatic acinar protein that stimulates pancreatic duct cell proliferation in vitro and islet neogenesis in vivo. To date, the mitogenic activity of INGAP has been identified only in nonneural tissues. The aim of this study was to examine the effects of a pentadecapeptide of INGAP (INGAP peptide), the biologically active portion of the native protein, in cultured dorsal root ganglia (DRG) explants from C57BL/6 mice. The present study provides evidence that INGAP peptide acts as a mitogen in the peripheral nervous system (PNS), and that it enhances neurite outgrowth from DRGs in vitro in a time- and dose-dependent manner. The neuritogenic action of INGAP peptide correlates with an increase in [(3)H]thymidine incorporation (P < 0.0001) and mitochondrial activity (P < 0.001). Results from these studies suggest that INGAP peptide promotes Schwann cell proliferation in the DRG which releases trophic factors that promote neurite outgrowth. PMID- 11855840 TI - Characterization of a Smad motif similar to Drosophila mad in the mouse Msx 1 promoter. AB - Mouse Msx 1 gene, orthologous of the Drosophila msh, is involved in several developmental processes. BMP family members are major proteins in the regulation of Msx 1 expression. BMP signaling activates Smad 1/5/8 proteins, which associate to Smad 4 before translocating to the nucleus. Analysis of Msx 1 promoter revealed the presence of three elements similar to the consensus established for Mad, the Smad 1 Drosophila counterpart. Notably, such an element was identified in an enhancer important for Msx 1 regulation. Gel shift analysis demonstrated that proteins from 13.5 dpc embryo associate to this enhancer. Remarkably, supershift assays showed that Smad proteins are present in the complex. Purified Smad 1 and 4 also bind to this fragment. We demonstrate that functional binding sites in this enhancer are confined to the Mad motif and flanking region. Our data suggest that this Mad motif may be functional in response to BMP signaling. PMID- 11855841 TI - Regulation of cyclic AMP-dependent response element-binding protein (CREB) by the nociceptin/orphanin FQ in human dopaminergic SH-SY5Y cells. AB - Nociceptin/orphanin FQ (N/OFQ), an endogenous ligand for opioid receptor-like (ORL1) receptor, transduces signaling cascades implicated in MAPK, PKC, PLC, and calcium, etc. This study was designed to investigate the intracellular signaling mechanism of N/OFQ in human dopaminergic neuroblastoma SH-SY5Y cells. N/OFQ rapidly induced the phosphorylation of CREB, which was significantly suppressed by pretreatment of PKA inhibitor, but not by MAPK inhibitors. It also time dependently increased the phosphorylation of MAPK, which was proven as ERKs, whereas it did not affect the PI3K activity. Interestingly, KT5720, a specific inhibitor of PKA, markedly suppressed the phosphorylation of MAPK by N/OFQ in SH SY5Y cells. Furthermore, BAPTA-AM, an intracellular chelator of Ca(2+), completely abolished the phosphorylation of CREB as well as MAPK in N/OFQ-treated SH-SY5Y cells. Taken together, these results suggest that N/OFQ independently induces the activation of CREB prior to MAPK phosphorylation, which was also modulated by PKA. Furthermore, Ca(2+)-related signaling implicates in the phosphorylation processes of CREB and MAPK simultaneously. PMID- 11855842 TI - Synergistic effect of cold mannitol and Na(+)/Ca(2+) exchange blocker on blood brain barrier opening. AB - The precise mechanism of reversible opening of the blood-brain barrier (BBB) is elusive. Hyperosmotic mannitol at 1.4 to 1.6 M is commonly used for this purpose but intraarterial injection of such a hyperosmotic solution is known to have possible side effects on the brain. Cooling of the solution has recently been reported to have potential to open the BBB. The in situ brain perfusion technique with [(14)C]-sucrose was used first to quantify BBB permeability in rats after intraarterial injection of 1.1 M mannitol, which is less hyperosmotic than commonly used mannitol, at room temperature and at 4 degrees C. Mannitol at 4 degrees C produced an opening of the BBB but the duration of the opening was less than 30 min. We then investigated the effect of an Na(+)/Ca(2+) exchange blocker (KB-R7943) on this hypothermic BBB opening. KB-R7943 extended the BBB opening to 30 min without affecting the peak level of BBB permeability at 5 min. Simple manipulation of temperature can thus enhance the reversible BBB opening in mannitol and pharmacological manipulation of calcium dynamics works synergistically with hypothermic mannitol. PMID- 11855843 TI - Atrophic change of rat salivary gland during adenovirus-induced hyperleptinemia. AB - Sustained hyperleptinemia in normal rats induced by infusing a recombinant adenovirus containing the rat leptin cDNA (AdCMV-leptin) exhibited a remarkable reduction in food intake (AdCMV-leptin, 9.3 +/- 2.6 vs untreated, 20.6 +/- 1.0 g/day) and ablated body fat without any significant changes in wet weight of liver and left ventricle. In those hyperleptinemic rats, we found a 52% reduction in wet weight of salivary gland compared with that in the pair-fed AdCMV-beta-gal treated rats, which received a recombinant virus containing the beta galactosidase gene (AdCMV-beta-gal) and were fed on the same amount of food as had been consumed by the AdCMV-leptin-treated group on the previous day. Microscopic examination with hematoxylin-eosin staining revealed that atrophic change was induced in both serous and mucous gland only in the AdCMV-leptin treated group, but not in the pair-fed controls. Thus, the atrophic changes in hyperleptinemic rats were due to neither a decrease of food intake nor disuse of the salivary gland related with anorexia. Our data suggested that size of the salivary gland was controlled, at lease in part, by "non-anorexic" effect of leptin. PMID- 11855844 TI - Bisphosphonates pamidronate and zoledronic acid stimulate osteoprotegerin production by primary human osteoblasts. AB - Bisphosphonates are potent antiresorptive drugs commonly employed in the treatment of metabolic bone diseases. Despite their frequent use, the mechanisms of bisphosphonates on bone cells have largely remained unclear. Receptor activator of nuclear factor-kappaB ligand (RANKL) is essential for osteoclast formation and activation, whereas osteoprotegerin (OPG) neutralizes RANKL. Various osteotropic drugs have been demonstrated to modulate osteoblastic production of RANKL and OPG. In this study, we assessed the effects of the bisphosphonates pamidronate (PAM) and zoledronic acid (ZOL) on OPG mRNA steady state levels (by semiquantitative RT-PCR) and protein production (by ELISA) in primary human osteoblasts (hOB). PAM increased OPG mRNA levels and protein secretion by hOB by up to 2- to 3-fold in a dose-dependent fashion with a maximum effect at 10(-6) M (P < 0.001) after 72 h. Similarly, ZOL enhanced OPG gene expression and protein secretion by hOB in a dose-dependent fashion with a maximum effect at 10(-8) M after 72 h, consistent with the higher biological potency of ZOL. Time course experiments indicated a stimulatory effect of PAM and ZOL on osteoblastic OPG protein secretion by 6-fold, respectively (P < 0.001). Pretreatment with PAM and ZOL prevented the inhibitory effects of the glucocorticoid dexamethasone on OPG mRNA and protein production. Analysis of cellular markers of osteoblastic differentiation revealed that PAM and ZOL induced type I collagen secretion and alkaline phosphatase activity by 2- and 4 fold, respectively (P < 0.0001 by ANOVA). In conclusion, our data suggest that bisphosphonates modulate OPG production by normal human osteoblasts, which may contribute to the inhibition of osteoclastic bone resorption. Since, OPG production increases with osteoblastic cell maturation, enhancement of OPG by bisphosphonates could be related to their stimulatory effects on osteoblastic differentiation. PMID- 11855845 TI - Lysophosphatidic acid stimulates p21-activated kinase in vascular smooth muscle cells. AB - Lysophosphatidic acid (LPA) has been shown to be a potent mitogen for vascular smooth muscle cells. Src-dependent transactivation of receptor tyrosine kinases has been previously demonstrated to mediate LPA-induced activation of MAP kinase ERK1/2. Furthermore, generation of reactive oxygen species (ROS) by LPA is also known to contribute to MAP kinase activation. Rho family small G-proteins Rac and Cdc42, and their immediate downstream effector p21-activated kinase (PAK), have been demonstrated to mediate important effects on the cytoskeleton that are relevant for cell migration and proliferation. In the present report we evaluated stimulation of PAK by LPA in rat aortic vascular smooth muscle cells (VSMC) by PAK immunocomplex MBP in-gel kinase assay. LPA increased PAK activity 3-fold, peaking at 5 min and showing sustained activation up to 45 min. Inhibition of tyrosine kinases by pretreatment of VSMC with genistein or specific inhibition of Src by PP1 greatly diminished LPA-induced PAK activation, whereas specific inhibition of PDFG- and EGF receptor kinase by tyrphostin AG1296 and AG1478 had no effect. Furthermore, inhibition of Galpha(i) by pertussis toxin and inhibition of NADH/NADPH oxidase by diphenylene iodonium also diminished LPA-induced stimulation of PAK. This is the first study to demonstrate that LPA activates PAK. In VSMC, PAK activation by LPA is mediated by Galpha(i) and is dependent on Src, whereas EGF- or PDGF receptor transactivation are not involved. Furthermore, generation of ROS is required for LPA-induced activation of PAK. PMID- 11855846 TI - Identification of an endocrine disrupting agent from corn with mitogenic activity. AB - A mitogenic agent in corncob bedding and fresh corn products disrupts sexual behavior and estrous cyclicity in rats. The mitogenic activity resides in an isomeric mixture of linoleic acid derivatives with a tetrahydrofuran ring and two hydroxyl groups (THF-diols) that include 9, (12)-oxy-10,13-dihydroxystearic acid and 10, (13)-oxy-9,12-dihydroxystearic acid. Synthetic THF-diols stimulated breast cancer cell proliferation in vitro and disrupted the estrous cycle in female rats at oral doses of approximately 0.30 mg/kg body weight/day. Exposure to THF-diols may disrupt endocrine function in experimental animals at doses approximately 200 times lower than classical phytoestrogens, promote proliferation of breast or prostate cancer, and adversely affect human health. PMID- 11855847 TI - Myostatin knockout in mice increases myogenesis and decreases adipogenesis. AB - Growth differentiation factor-8 (GDF-8), or Myostatin, plays an important inhibitory role during muscle development. Since muscle and adipose tissue develop from the same mesenchymal stem cells, we hypothesized that Myostatin gene knockout may cause a switch between myogenesis and adipogenesis. Male and female wild type (WT) and Myostatin knockout (KO) mice were sacrificed at 4, 8, and 12 weeks of age. The gluteus muscle (GM) was larger in KO mice compared to WT mice at 8 (P < 0.01) and 12 (P < 0.001) weeks. At 12 weeks, KO mice had decreased fat depots (P < 0.01). Compared to 12-week-old WT mice, serum leptin concentration in KO mice was lower (P < 0.001) and leptin mRNA expression was decreased (P < 0.01) in inguinal adipose tissue. CCAAT/enhancer binding protein-alpha (C/EBPalpha) and peroxisome proliferator-activated receptor-gamma (PPARgamma) levels in adipose tissue were significantly lower in KO mice compared to WT mice. Thus, increased muscle development in Myostatin knockout mice is associated with reduced adipogenesis and consequently, decreased leptin secretion. PMID- 11855848 TI - Mga2p is a putative sensor for low temperature and oxygen to induce OLE1 transcription in Saccharomyces cerevisiae. AB - Various low-temperature-inducible genes such as fatty acid desaturase genes are essential for all living organisms to acclimate to low temperature. However, a low-temperature signal transduction pathway has not been identified in eukaryotes. In yeast Saccharomyces cerevisiae, the Delta9 fatty acid desaturase gene OLE1 is activated by ubiquitin/proteasome-dependent processing of two homologous endoplasmic reticulum membrane proteins, Spt23p and Mga2p. We found that OLE1 transcription was transiently activated with resultant increases in the degree of unsaturation of total fatty acids when culture temperature was downshifted from 30 degrees C to 10 degrees C. This activation was greatly depressed in Deltamga2 cells. Although Mga2p is essential for hypoxic activation of OLE1 transcription, and its hypoxic functions are repressed by unsaturated fatty acids (UFAs), low-temperature activation of the OLE1 gene was not repressed by UFAs. These observations suggest that low-temperature and hypoxic signal transduction pathways share some components, and Mga2p is the first identified eukaryotic sensor for low temperature and oxygen. PMID- 11855849 TI - Induction of global stress response in Saccharomyces cerevisiae cells lacking telomerase. AB - Cellular senescence is a major intermediate step from healthy cells toward tumor cells. By using microarrays that simultaneously examine the transcription levels of 6,200 Saccharomyces cerevisiae genes, we show that 45 gene transcript levels are increased and 11 are decreased after exposure to telomere shortening and cellular senescence in a telomerase-deficient mutant. About half of the genes that showed increased expression were found induced under stress, consistent with the notion that critical short telomeres cause stress to cells. Surprisingly, the expression level of telomere recombination genes was not altered suggesting that even though recombination is a means to rescue critically short telomeres, its machinery was not controlled by telomere shortening. The expression of telomere proximal genes was also analyzed. The possibility of induction of a program to cope with cellular senescence and active telomere-telomere recombination is discussed. PMID- 11855851 TI - Evidence for repeat-induced gene silencing in cultured Mammalian cells: inactivation of tandem repeats of transfected genes. AB - Foreign DNA can be readily integrated into the genomes of mammalian embryonic cells by retroviral infection, DNA microinjection, and transfection protocols. However, the transgenic DNA is frequently not expressed or is expressed at levels far below expectation. In a number of organisms such as yeast, plants, Drosophila, and nematodes, silencing of transfected genes is triggered by the interaction between adjacent or dispersed copies of genes of identical sequence. We set out to determine whether a mechanism similar to repeat-induced gene silencing (RIGS) is responsible for the silencing of transgenes in murine embryonal carcinoma stem cells. We compared the expression of identical reporter gene constructs in cells carrying single or multiple copies and found that the level of expression per integrated copy was more than 10-fold higher in single copy integrants. In cells carrying tandem copies of the transgene, many copies were methylated and clones frequently failed to express both copies of near identical integrated alleles. Addition of extra copies of the reporter gene coding sequence reduced the level of expression from the same reporter driven by a eukaryotic promoter. We also found that inhibitors of histone deacetylase such as trichostatin A forestall the silencing of multicopy transgenes, suggesting that chromatin mediates the silencing of transfected genes. This evidence is consistent with the idea that RIGS does occur in mammalian embryonic stem cells although silencing of single-copy transgenes also occurs, suggesting that RIGS is only one of the mechanisms responsible for triggering transgene silencing. PMID- 11855850 TI - Metformin restores responses to insulin but not to growth hormone in Sprague Dawley rats. AB - Administration of growth hormone (GH) increases muscle mass in F344 x BN rats, but not in Sprague-Dawley (S-D) rats. S-D rats are insulin-resistant and insulin responsiveness is required for the anabolic actions of GH. We hypothesized that correction of insulin resistance with metformin might also restore anabolic effects of GH. Treatment with GH (0.25 or 1.0 mg/kg twice daily for 9 days) had limited anabolic effects, reducing weight gain by 14%, increasing muscle glycogen content by 40% and increasing exercise capacity by 24%, but failing to increase muscle mass or to reduce fat mass. GH also impaired insulin responsiveness and increased visceral fat TNF content of visceral fat by 77%. Metformin enhanced insulin responsiveness in skeletal muscle, but failed to enhance anabolic effects of GH. Rats aged 14 weeks were treated for 21 days with metformin (320 mg/kg/day) and for the last 9 days, with GH (0.25 mg/kg, twice daily). Metformin caused a 2.3-fold increase in insulin-stimulated muscle glucose transport and a 20% reduction in muscle fatty acid oxidation, indicating increased glucose utilization. However, metformin did not augment GH-induced weight reduction. Metformin decreased visceral fat by 22% and subcutaneous fat by 20%, but no decreases were observed in the GH/metformin group. GH increased muscle glycogen by 40%, but the effect was reversed by metformin. VO(2max) was increased 24% by GH and 17% by metformin, but was not elevated in the GH/metformin group. GH increased TNF in visceral fat and the effect was augmented by metformin (144% increase). We conclude that metformin enhances some aspects of insulin responsiveness, but does not enhance anabolic responses to GH. The latter may, in part, be explained by the failure of metformin to prevent GH-induced elevation of TNF in visceral fat. PMID- 11855852 TI - Decreased apoptotic response of inclusion-cell disease fibroblasts: a consequence of lysosomal enzyme missorting? AB - To better understand the role of lysosomes in apoptosis, we compared the responses to apoptotic stimuli of normal fibroblasts with those of inclusion cells (I-cells), i.e., fibroblasts with impaired function of lysosomal enzymes due to their missorting and ensuing nonlysosomal localization. Although both cell types did undergo apoptosis when exposed to the lysosomotropic detergent MSDH, the redox-cycling quinone naphthazarin, or the protein kinase inhibitor staurosporine, I-cells exerted a markedly decreased response to these agonists than did normal fibroblasts. Furthermore, leupeptin and pepstatin A (inhibitors of cysteine and aspartic proteases, respectively) suppressed staurosporine induced apoptosis of normal fibroblasts, whereas survival of I-cells was unaffected. These findings give further support for the involvement of lysosomal enzymes in apoptosis and suggest I-cells as a suitable model for studying the role of lysosomes in programmed cell death. PMID- 11855853 TI - Role of the mitochondrial permeability transition and cytochrome C release in hydrogen peroxide-induced apoptosis. AB - We investigated the role of the mitochondrial inner membrane permeability transition and subsequent release of cytochrome c into the cytosol during oxidative stress-evoked apoptosis. Sublethal oxidative stress was applied by treating L929 cells with 0.5 mM H2O2 for 90 min. Then the cellular localization of cytochrome c was examined by immunofluorescent staining and Western blotting. H2O2 treatment caused the permeability transition and pore formation, resulting in membrane depolarization and translocation of cytochrome c from the mitochondria into the cytosol. Pretreatment with cyclosporin A and aristolochic acid (to inhibit pore formation) significantly attenuated a reduction of the mitochondrial membrane potential, as well as signs of apoptosis such as DNA fragmentation, increased plasma membrane permeability, and chromatin condensation. Therefore, exposure to H2O2 caused the opening of permeability transition pores in the inner mitochondrial membrane. An essential role of cytosolic cytochrome c in the execution of apoptosis was demonstrated by its direct microinjection into the cytosol, thus bypassing the need for cytochrome c release from the mitochondrial intermembrane space. Microinjection of cytochrome c caused caspase-dependent apoptosis. PMID- 11855854 TI - The histone deacetylase inhibitor trichostatin A derepresses the telomerase reverse transcriptase (hTERT) gene in human cells. AB - Activation of telomerase, essential for cellular immortalization and transformation, requires the induction of its catalytic component, telomerase reverse transcriptase (hTERT). However, biochemical and genetic mechanisms for the control of hTERT expression remain undefined. In the present study, we demonstrate that the histone deacetylase (HDAC) inhibitor trichostatin A (TSA) induces hyperacetylation of histones at the hTERT proximal promoter, directly transactivates the hTERT gene in normal human telomerase-negative cells, and upregulates hTERT expression in telomerase-positive tumor cells. Overexpression of HDAC1 leads to repression of the hTERT promoter activity. TSA-mediated activation of the hTERT promoter is abolished by the mutation of Sp1 sites at the proximal promoter, suggesting that the effect of TSA is regulated through Sp1 motifs. We also show a physical interaction of Sp1 with HDAC1 and the presence of HDAC1 at the hTERT promoter region. Moreover, hyperacetylation of histones at the hTERT promoter is associated with the natural up-regulation of hTERT expression that occurs in activated T lymphocytes. Taken together, histone acetylation/deacetylation may be a common underlying feature to hTERT transactivation/repression in human normal and malignant cells. PMID- 11855855 TI - An octapeptide in the juxtamembrane domain of VE-cadherin is important for p120ctn binding and cell proliferation. AB - The cadherins are a family of adhesive proteins involved in cell-cell homophilic interactions. VE-cadherin, expressed in endothelial cells, is involved in morphogenesis, regulation of permeability, and cellular proliferation. The cytoplasmic tails of cadherins contain two major domains, the juxtamembrane domain that plays a role in the intercellular localization of the protein and also serves for binding of p120ctn, and a C-terminal domain that associates with beta- or gamma-catenin. A highly conserved region present in the juxtamembrane domain of the cadherins has been shown to be necessary for p120ctn binding in E cadherin. Using a mutant VE-cadherin lacking a highly conserved octapeptide, we demonstrated that it is required for p120ctn binding to VE-cadherin as determined by immunoprecipitation and colocalization studies. By immunofluorescence, this mutant protein has a topographical distribution similar to that of the wild-type VE-cadherin and, therefore, we conclude that the topographical distribution of VE cadherin is independent of this motif. In addition, although cell-cell association is present in cells expressing this mutant form of VE-cadherin, we found that the strength of adhesion is decreased. Finally, our results for the first time demonstrate that the interaction of VE-cadherin with p120 catenin plays an important role in cellular growth, suggesting that the binding of p120 catenin to cadherins may regulate cell proliferation. PMID- 11855856 TI - Synergistic activation of the TATA-less mouse thymidylate synthase promoter by the Ets transcription factor GABP and Sp1. AB - The mouse thymidylate synthase (TS) promoter lacks a TATA box and an initiator element and directs transcriptional initiation at multiple sites over a 90 nucleotide region. The minimum sequence required for wild-type promoter activity has been mapped to a 30-nucleotide essential promoter region that partially overlaps the 5' end of the transcriptional initiation window. The essential promoter region contains two potential binding sites for members of the Ets family of transcription factors as well as a binding site for Sp1. Promoter mutation analyses revealed that all three of these sites are important for promoter activity. Transient cotransfection assays showed that GABP, a heterodimeric Ets factor, is able to stimulate expression of reporter genes driven by the wild-type mouse TS promoter whereas several other Ets factors have no effect. Electrophoretic mobility shift assays revealed that recombinant GABP binds to both Ets elements in the essential promoter region. Stimulation of promoter activity by GABP is diminished when either Ets element is inactivated and is prevented when both Ets elements are inactivated. Transient cotransfection assays revealed that Sp1 and GABP stimulate TS promoter activity in a highly synergistic manner. PMID- 11855857 TI - Structural polarity and functional bile canaliculi in rat hepatocyte spheroids. AB - Primary hepatocytes self-assemble into spheroids that possess tight junctions and microvilli-lined channels. We hypothesized that polarity develops gradually and that the channels structurally and functionally resemble bile canaliculi. Immunofluorescence labeling of apical and basolateral proteins demonstrated reorganization of the membrane proteins into a polarized distribution during spheroid culture. By means of fluorescent dextran diffusion and confocal microscopy, an extensive network of channels was revealed in the interior of the spheroids. These channels connected over several planes and opened to pores on the surface. To examine the content of apical proteins in the channel membranes, the bile canalicular enzyme dipeptidyl peptidase IV (DPPIV) was localized using a fluorogenic substrate, Ala-Pro-cresyl violet. The results show that DPPIV activity is heterogeneously distributed in spheroids and localized in part to channels. Bile acid excretion was then investigated to demonstrate functional polarity. A fluorescent bile acid analogue, fluorescein isothiocyanate-labeled glycocholate, was taken up into the spheroids and excreted into bile canalicular channels. Due to the structural polarity of spheroids and their ability to excrete bile into channels, they are a unique three-dimensional model of in vitro liver tissue self-assembly. (Videoanimations of some results are available at http://hugroup.cems.umn.edu/research_movies). PMID- 11855858 TI - Motility and ramification of human fetal microglia in culture: an investigation using time-lapse video microscopy and image analysis. AB - Microglia are mononuclear phagocytes of the central nervous system and are considered to derive from circulating bone marrow progenitors that colonize the developing human nervous system in the second trimester. They first appear as ameboid forms and progressively differentiate to process-bearing "ramified" forms with maturation. Signals driving this transformation are known to be partly derived from astrocytes. In this investigation we have used cocultures of astrocytes and microglia to demonstrate the relationship between motility and morphology of microglia associated with signals derived from astrocytes. Analysis of progressive cultures using time-lapse video microscopy clearly demonstrates the dynamic nature of microglia. We observe that ameboid microglial cells progressively ramify when cocultured with astrocytes, mirroring the "differentiation" of microglia in situ during development. We further demonstrate that individual cells undergo morphological transformations from "ramified" to "bipolar" to "tripolar" and "ameboid" states in accordance with local environmental cues associated with astrocytes in subconfluent cultures. Remarkably, cells are still capable of migration at velocities of 20-35 microm/h in a fully ramified state overlying confluent astrocytes, as determined by image analysis of motility. This is in keeping with the capacity of microglia for a rapid response to inflammatory cues in the CNS. We also demonstrate selective expression of the chemokines MIP-1alpha and MCP-1 by confluent human fetal astrocytes in cocultures and propose a role for these chemotactic cytokines as regulators of microglial motility and differentiation. The interchangeable morphological continuum of microglia supports the view that these cells represent a single heterogeneous population of resident mononuclear phagocytes capable of marked plasticity. PMID- 11855859 TI - Mechanical stress is required for high-level expression of connective tissue growth factor. AB - We used gene array technology to analyze differences in gene expression between mechanically stressed and relaxed fibroblasts. A number of stress-responsive genes that showed a two- to sixfold difference in their relative expression were identified. Connective tissue growth factor (CTGF) was among those genes that showed the most striking up-regulation by mechanical stress. Its regulation occurred at the transcriptional level and was reversible. A new steady state level of CTGF mRNA was reached within less than 6 h after stress relaxation. Mechanical stress was absolutely required for sustained high-level expression; TGF-beta, which is also known to stimulate CTGF synthesis, was not sufficient on its own. Experiments with specific inhibitors suggested that a protein kinase and a tyrosine phosphatase were involved in the transduction of the mechanical stimulus to gene expression. Since CTGF controls the synthesis of several extracellular matrix proteins, it is likely that this growth factor is responsible for the increased synthesis of collagen I and other matrix proteins in stressed fibroblasts. PMID- 11855861 TI - Clofibrate-induced relocation of phosphatidylcholine transfer protein to mitochondria in endothelial cells. AB - The phosphatidylcholine transfer protein (PC-TP) is a specific transporter of phosphatidylcholine (PC) between membranes. To get more insight into its physiological function, we have studied the localization of PC-TP by microinjection of fluorescently labeled PC-TP in foetal bovine heart endothelial (FBHE) cells and by expression of an enhanced yellow fluorescent protein-PC-TP fusion protein in FBHE cells, human umbilical vein endothelial cells, and HepG2 cells. Analysis by confocal laser scanning microscopy showed that PC-TP was evenly distributed throughout the cytosol with an apparently elevated level in nuclei. By measuring the fluorescence recovery after bleaching it was established that PC-TP is highly mobile throughout the cell, with its transport into the nucleus being hindered by the nuclear envelope. Given the proposed function of PC TP in lipid metabolism, we have tested a number of compounds (phorbol ester, bombesin, A23187, thrombin, dibutyryl cyclic AMP, oleate, clofibrate, platelet derived growth factor, epidermal growth factor, and hydrogen peroxide) for their ability to affect intracellular PC-TP distribution. Only clofibrate (100 microM) was found to have an effect, with PC-TP moving to mitochondria within 5 min of stimulation. This relocation did not occur with PC-TP(S110A), lacking the putative protein kinase C (PKC)-dependent phosphorylation site, and was restricted to the primary endothelial cells. Relocation did not occur in HepG2 cells, possibly due to the fact that clofibrate does not induce PKC activation in these cells. PMID- 11855860 TI - Induction of apoptosis in human replicative senescent fibroblasts. AB - Cellular senescence is an irreversible growth phase characteristic of normal cells. We have found that human senescent fibroblasts can be induced to undergo programmed cell death (apoptosis) by ceramide, TNF-alpha, or okadaic acid. The most profound effects were induced by TNF-alpha and okadaic acid treatment. In the present study, we also evaluated the contribution of lysosomal activation as a possible mechanism underlying the induction of apoptosis. Four lysosomal enzyme activities were measured: beta-galactosidase, alpha-galactosidase A, beta glucoronidase, and acid phosphatase. Using an in situ assay, we have found that the activity of beta-galactosidase, which is also a biochemical marker of senescence, is induced in young proliferating fibroblasts following exposure to all three apoptotic inducing agents. The other enzymes were not significantly induced in young fibroblasts following exposure to agents that induce apoptosis. During replicative senescence, three of the four lysosomal enzymes tested (beta galactosidase, alpha-galactosidase A, and beta-glucoronidase) are constitutively expressed at high levels. TNF-alpha was the only agent that induced lysosomal activity in senescent fibroblasts, of which only alpha-galactosidase A activity was induced. Our studies show that senescent fibroblasts can be induced to undergo apoptosis in a signal-dependent manner. However, the lysosomal enzymes examined do not appear to be correlated with apoptotic induction. PMID- 11855862 TI - Processes that occur before second cleavage determine third cleavage orientation in Xenopus. AB - As in many organisms, the first three cleavage planes of Xenopus laevis eggs form in a well-described mutually orthogonal geometry. The factors dictating this simple pattern have not been unambiguously identified. Here, we describe experiments, using static magnetic fields as a novel approach to perturb normal cleavage geometry, that provide new insight into these factors. We show that a magnetic field applied during either or both of the first two cell cycles can induce the third cell cycle mitotic apparatus (MA) at metaphase and the third cleavage plane to align nearly perpendicular to their nominal orientations without changing cell shape. These results indicate that processes occurring during the first two cell cycles primarily dictate the third cleavage plane and mitotic apparatus orientation. We discuss how mechanisms that can align the MA after it has formed are likely to be of secondary importance in determining cleavage geometry in this system. PMID- 11855864 TI - Retinoid signaling is attenuated by proteasome-mediated degradation of retinoid receptors in human keratinocyte HaCaT cells. AB - The biological actions of retinoids are mediated by nuclear retinoid receptors, RAR and RXR, which are ligand-activated transcription factors. We investigated the mechanism of attenuation of retinoid receptor activity in human keratinocyte HaCaT cells. Treatment of HaCaT cells with all-trans-retinoic acid or 9-cis retinoic acid reduced RARgamma and RXRalpha protein levels by one-half within 24 h. In contrast, retinoid treatment did not alter RARgamma or RXRalpha mRNA levels, suggesting that retinoids stimulate breakdown of their receptors. Pulse chase studies revealed that retinoid treatment of HaCaT cells reduced RARgamma and RXRalpha half-lives by 50%, indicating that retinoids accelerate breakdown of their receptors. The proteasome inhibitor MG132 prevented retinoid-induced receptor loss. Furthermore, MG132 potentiated retinoid-induced receptor activity, as assessed by expression of the retinoid-regulated CRABP-II gene in HaCaT cells. These data demonstrate that retinoids attenuate retinoid receptor function by enhancing proteasome-mediated retinoid receptor breakdown in HaCaT cells. Proteasome-mediated degradation of RARgamma or RXRalpha in vitro was significantly reduced by the corepressor SMRT, which binds unliganded retinoid receptors. This protection from degradation was markedly diminished by ligand, which causes SMRT to dissociate from receptors. Ligand failed to relieve protection from degradation by SMRT of a mutant form of RXRalpha that binds SMRT in the presence and absence of ligand. Addition of coactivators TIF1, TIF2, and RIP140 had no effect on degradation of RARgamma or RXRalpha. In summary, ligand binding to retinoid receptors promotes proteasome-mediated receptor degradation via dissociation of SMRT. Ligand-stimulated receptor degradation results in attenuation of retinoid signaling. PMID- 11855863 TI - LIF-induced STAT3 signaling in murine versus human embryonal carcinoma (EC) cells. AB - Self-renewal and the maintenance of pluripotency of mouse embryonal stem (ES) cells in vitro requires exogenous leukemia inhibitory factor (LIF). Mouse ES cells can be cultured and kept undifferentiated in the absence of embryonal feeder-cell layers when exogenous LIF concentrations are maintained above a threshold concentration. An important downstream target of LIF signal transduction in mouse ES cells is the transcription factor signal transducer and activator of transcription 3 (STAT3). In contrast to mouse ES cells, human ES cells are unresponsive to LIF and depend on feeder cells for undifferentiated growth. Here, we investigated the activation patterns of LIF-downstream effectors in mouse and human embryonal carcinoma (EC) cells. We report that LIF induces both ERK-1 as well as STAT3 activation in mouse P19 EC cells. LIF enhances the proliferation rate of P19 EC cells, which depends on ERK activity but does not require activation of STAT3. In contrast, LIF does not activate STAT3, ERK, or the gp130 receptor in human N tera-2/D1 EC cells, although all receptor components are expressed. The negative feedback protein suppressor of cytokine signaling 1 (SOCS-1) is constitutively expressed in N tera-2/D1 EC cells, suggesting that LIF signal transduction is inhibited by elevated levels of SOCS-1 expression. PMID- 11855865 TI - Nuclear localization of the tight junction protein ZO-2 in epithelial cells. AB - The tight junction constitutes the major barrier to solute and water flow through the paracellular space of epithelia and endothelia. It is formed by transmembrane proteins and submembranous molecules such as the MAGUKs ZOs. We have previously found that several MAGUKs, including those of the tight (ZO-1, ZO-2, and ZO-3) and septate junction (tamou and Dlg), contain one or two nuclear sorting signals located at their first PDZ and GK domains. Now we show that these proteins also contain a nuclear export signal and focus our study on the nuclear membrane shuttling of ZO-2. In sparse cultures this molecule concentrates at the nucleus in clusters, where it partially colocalizes with splicing factor SC35. Nuclear staining diminishes as the monolayer acquires confluence through a process sensitive to the nuclear export inhibitor leptomycin B. Nuclear localization can be induced by impairing cell-cell contacts, by mechanical injury. ZO-2 that shuttles from the cell periphery into the nucleus is not newly synthesized but originates from a preexistent pool. The movement of this protein is mediated by the actin cytoskeleton. PMID- 11855866 TI - Dissection of protease-activated receptor-1-dependent and -independent responses to thrombin in skeletal myoblasts. AB - Thrombin exerts a number of effects on skeletal myoblasts in vitro. It stimulates proliferation and intracellular calcium mobilization and inhibits differentiation and apoptosis induced by serum deprivation in these cells. Many cellular responses to thrombin are mediated by protease-activated receptor-1 (PAR-1). Expression of PAR-1 is present in mononuclear myoblasts in vitro, but repressed when fusion occurs to form myotubes. In the current study, we used PAR-1-null mice to determine which of thrombin's effects on myoblasts are mediated by PAR-1. Thrombin inhibited fusion almost as effectively in cultures prepared from the muscle of PAR-1-null myoblasts as in cultures prepared from wild-type mice. Apoptosis was inhibited as effectively in PAR-1-null myoblasts as in wild-type myoblasts. These effects in PAR-1-null myoblasts were mediated by a secreted inhibitor of apoptosis and fusion, as demonstrated previously for normal rat myoblasts. Thrombin failed to induce an intracellular calcium response in PAR-1 null myoblast cultures, although these cells were able to mobilize intracellular calcium in response to activation of other receptors. PAR-1-null myoblasts also failed to proliferate in response to thrombin. These results demonstrate that thrombin's effects on myoblast apoptosis and fusion are not mediated by PAR-1 and that PAR-1 is the only thrombin receptor capable of inducing proliferation and calcium mobilization in neonatal mouse myoblasts. PMID- 11855867 TI - Activation of NF-kappaB is required for PDGF-B chain to transform NIH3T3 cells. AB - Elucidating the secondary signaling molecules that are necessary for platelet derived growth factor (PDGF) to stimulate tumor development will be crucial to the understanding and treatment of a variety of cancers. Several lines of evidence have indicated that the transcription factor NF-kappaB plays a central role in transformation induced by Ha-ras and Bcr-abl, but nothing is known concerning its role in transformation by PDGF. Here we demonstrate that transcription from a promoter containing NF-kappaB binding sequences as well as the DNA binding activity of NF-kappaB were increased in PDGF-B-chain-transformed mouse fibroblast cells. Focus formation of PDGF-B-chain-transformed mouse fibroblasts was suppressed by treatment with acetylsalicylic acid (ASA) and salicylic acid, which are known inhibitors of NF-kappaB activation, but other nonsteroidal anti-inflammatory drugs that do not have an effect on NF-kappaB activity did not affect focus formation in these cells. Furthermore, expression of a dominant negative mutant of IkappaBalpha, pMEIkappaBalpha67CJ, and a dominant negative mutant of p65, p65DeltaC, resulted in decreased focus formation and NF-kappaB activity. Therefore, the transcription factor NF-kappaB plays a vital role in PDGF-B chain transformation of mouse fibroblast cells, and the NF kappaB activity is sensitive to treatment with ASA. PMID- 11855868 TI - Surgery "ain't" going to cut it. Are we ready? PMID- 11855869 TI - Complementary and alternative therapy use in gynecologic oncology: implications for clinical practice. PMID- 11855870 TI - Use of complementary and alternative medicine among women with gynecologic cancers. AB - OBJECTIVE: The aim of this study was to determine the prevalence and types of complementary and alternative medicine (CAM) usage by women with gynecologic cancer in an outpatient midwestern university practice. METHODS: Any patient with a gynecologic cancer seen in the outpatient clinic of the gynecologic oncology division at Washington University over a 3-month period was eligible, excluding those patients with a new cancer diagnosis. Subjects completed a questionnaire anonymously. Two by two comparisons were made using the Fisher exact test and P was considered significant at P < 0.05. RESULTS: Nearly half (49.6%) of 113 respondents had used CAM since being diagnosed with cancer. Characteristics significantly associated with CAM use include annual income greater than $30,000, cancer site of origin other than the cervix, and use of CAM prior to cancer diagnosis. Users with annual incomes greater than $30,000 were significantly more likely to use CAM in the "other" category that included acupuncture, reflexology, and electromagnetic therapy. Fewer than 25% of CAM users received information regarding CAM from a physician, nurse, or practitioner of CAM. Women used CAM in hopes of achieving a wide range of potential benefits including both improved well-being and anti-cancer effects. The most common actual benefit these women perceived was an improvement in psychosocial well-being, including increased hope or optimism. CONCLUSIONS: American patients with gynecologic cancer frequently use CAM in addition to standard medical therapy. Oncologists caring for women with gynecologic cancer should initiate a dialogue about usage of CAM, discussing the potential adverse effects of CAM and the patient's therapeutic goals. PMID- 11855871 TI - Ifosfamide and vinorelbine in advanced platinum-resistant ovarian cancer: excessive toxicity with a potentially active regimen. AB - OBJECTIVES: The aim of this study was to determine the activity and toxicity of a vinorelbine and ifosfamide combination in platinum-resistant advanced ovarian cancer. PATIENTS AND METHODS: Patients were treated with ifosfamide (2 g/m(2)/day) infused over 1 h x 3 days (with mesna uroprotection) and vinorelbine (30 mg/m(2)) on days 1 and 8. Treatment was repeated on a 21-day schedule. In order to avoid unacceptable toxicity in this subset of patients where the chemotherapy is mainly palliative, the Bryant and Day two-stage phase II trial design incorporating toxicity considerations was chosen. A cutoff point for the response rate (10%) and for severe toxicity (25%) was established for the first 14 patients. RESULTS: Between February 1997 and December 1998, 11 paclitaxel and platinum-resistant patients and 1 potentially platinum-sensitive patient were treated. Five patients (41%) experienced grade 3-4 central nervous toxicity requiring hospital admission. In accordance with the Bryant and Day design, the study was stopped early because greater than 25% of the first 14 patients developed grade 3-4 neurotoxicity. A retrospective review of clinical characteristics of these patients showed at least one well-known risk factor associated with ifosfamide central toxicity. Hematological toxicity was common, mainly grade 4 neutropenia, which was observed in all but 1 patient, usually of short duration, and there were 4 episodes of neutropenic fever. Ten patients were evaluated for response. Two complete responses and 1 partial response according to CA-125 criteria were observed. CONCLUSION: This combination may be active in platinum-resistant ovarian cancer but the high toxicity encountered, principally neurotoxicity in those with large central pelvic masses, means that further studies with this schedule may not be warranted. PMID- 11855872 TI - Antibody levels against galactosyl (alpha1 --> 3) galactose epitopes in cervical mucus from patients with human papillomavirus infection. AB - OBJECTIVES: The general objective of the present study is to quantify antigalactosyl (alpha1 --> 3) galactose (anti-Gal) antibody levels in the cervical mucus of patients with/without human papillomavirus (HPV) infection and, as specific objectives, to compare these levels in the different HPV subgroups and with the presence of intraepithelial lesions. METHODS: Sixty women between 18 and 35 years old (mean: 26 years) were studied through the following methods: vaginal cytology, colposcopy, biopsy of suspicious lesions, sample taking for evaluating HPV presence through PCR and hybridization, and quantification of anti Gal levels using ELISA with laminin antigen extracted from the Engelbreth-Horm Swarm cell line. RESULTS: The presence of HPV was detected in 55% of patients; 45% of them had intermediate/high oncogenic risk HPV, 12% had low oncogenic risk HPV, and the other 43% had both subgroups. There were significantly higher anti Gal levels in the HPV+ group when compared with the HPV- group (P < 0.0001); also, when dividing the HPV+ group into one subgroup with normal cytology and another with cervical intraepithelial neoplasia (CIN) 1, we found higher values in the latter group (P < 0.0001). There was no significant difference in anti-Gal levels in the various HPV subgroups. CONCLUSION: High anti-Gal levels are found in the cervical mucus of patients with HPV infection and CIN 1, which suggests the participation of local humoral immunity in cervical lesions. PMID- 11855873 TI - Analysis of patients with epithelial ovarian cancer or fallopian tube carcinoma retreated with cisplatin after the development of a carboplatin allergy. AB - OBJECTIVE: We report the outcome of seven patients treated for recurrent ovarian cancer with cisplatin after an allergic reaction to carboplatin. One case is presented in which a heavily pretreated patient suffered a severe anaphylactic reaction, which was refractory to standard resuscitative measures and resulted in her death. METHODS: Six further patients who received cisplatin after documentation of an allergic reaction to carboplatin (CBDCA) for the treatment of recurrent epithelial ovarian cancer between 1993 and 2000 were identified from the MSKCC database. Electronic medical records were reviewed for relevant treatment and outcome data. RESULTS: Five of six of these patients were successfully treated without further allergic reactions. One patient with platinum-refractory disease had an allergic reaction to carboplatin and subsequently to cisplatin. CONCLUSION: Few patients with a carboplatin allergy are subsequently treated with cisplatin at our center. One patient suffered a serious hypersensitivity reaction following retreatment and died. Based on this limited experience, cross allergy can exist although the true incidence is not known. Routine retreatment of carboplatin-allergic patients with cisplatin in the relapsed setting cannot be recommended without careful consideration of potential risks and benefits. PMID- 11855874 TI - Cervical intraepithelial neoplasia is associated with increased polyamine oxidase and diamine oxidase concentrations in cervical mucus. AB - OBJECTIVE: The aim of this study was to establish whether reactive oxygen species, generated during oxidation of amines, catalyzed by polyamine oxidase (PAO) and diamine oxidase (DAO) in cervical secretions may play a role in the etiology of cervical cancer. METHODS: Cervical mucus was obtained from women attending the gynecological outpatient department: 139 with and 154 without cytological evidence of cervical intraepithelial neoplasia were recruited. The mucus was freeze dried in liquid nitrogen, weighed, and later resuspended for assay of PAO and DAO concentrations using a chemiluminescence method. The two groups were compared by group sequential analysis using PEST3 software. RESULTS: Patients with a colposcopic diagnosis of a high-grade squamous intraepithelial lesion (SIL) had significantly higher enzyme activities than control cases (L(N)PAO 1.37 (0.37) versus 1.18 (0.35): Student t test: P < 0.001; L(N)DAO 1.37 (0.36) versus 1.15 (0.37): Student t test: P < 0.001). CONCLUSION: It is probable that this rise in enzyme activity precedes cytological changes and plays some part in the etiology of cervical cancer, as the cells that undergo premalignant change are normally squamous in origin, whereas mucus is a product of columnar epithelium. Higher enzyme activity in patients with SIL than in controls may be a reflection of higher risk of exposure to amine substrates in semen through multiple sexual partners. PMID- 11855875 TI - The impact of performance status on survival in patients of 80 years and older with vulvar cancer. AB - OBJECTIVE: There are no data available on the impact of performance status on outcome in patients with vulvar cancer. It was the objective of this study to determine the impact of performance status on survival in a group of elderly patients. METHODS: A retrospective review of records of patients with vulvar cancer aged 80 years or greater and treated in a gynecological referral center was performed. Multiple clinical and pathological variables together with performance status were assessed and the impact on overall survival was determined both by univariate and multivariate analysis. RESULTS: Of 75 patients aged 80 years or older, 57 (76%) had standard treatment. The patients who had standard treatment were characterized by an earlier clinical stage and a better performance status compared with patients who had nonstandard treatment. When preoperatively available parameters of all patients were assessed in relation to survival in the total group, Eastern Cooperative Oncology Group (ECOG) performance status was the only independent prognostic indicator for survival. When all clinical and histopathological variables were assessed in the subgroup who had standard treatment, both ECOG performance status and extracapsular lymph node involvement were independent prognostic variables for overall survival. Age was not a significant prognostic variable. CONCLUSIONS: ECOG performance status is the only available pretreatment variable with independent prognostic value for survival in this group of elderly patients with vulvar cancer. These data show the importance of individualizing the treatment of patients with vulvar cancer. Performance status takes a more important place than age in the management process of these patients. PMID- 11855876 TI - Hysteroscopic dissemination of endometrial carcinoma using carbon dioxide and normal saline: a retrospective study. AB - OBJECTIVE: The aim of this study was to compare the likelihood of disseminating endometrial carcinoma cells into the peritoneal cavity by hysteroscopic examination using carbon dioxide (CO(2)) or normal saline (NS) as the distension medium. METHODS: A retrospective study of 162 consecutive patients with endometrial carcinoma treated at a university teaching hospital from 1994 to 1999 was undertaken. All patients had a hysteroscopic examination, using either CO(2) or NS as the distension medium, as part of the investigation for abnormal uterine bleeding or in determining whether the uterine cervix was invaded by tumor. Peritoneal fluid for cytology was collected immediately upon entry into the abdominal cavity. Positive peritoneal cytology was considered the primary statistical endpoint. RESULTS: Among 162 patients, 39 cases were excluded from the study because of macroscopic intraperitoneal diseases (n = 32) or pathology other than endometrioid adenocarcinoma (n = 7). Another 3 cases were excluded because both distension mediums had been used in the hysteroscopy. Analysis was therefore based on the data of 120 patients. There was no statistically significant difference between the two groups of patients undergoing hysteroscopy using either CO(2) (n = 70) or NS (n = 50) with regard to age, pathologic stage, International Federation of Gynecology and Obstetrics grading, myometrial invasion, tumor size, cervical involvement, nodal involvement, and 2-year progression-free survival. However, there was a mean of 13.0 plus minus 5.0 days (range 3-21 days) time gap between laparotomy for definitive surgery and CO(2) hysteroscopy compared to immediate laparotomy after NS hysteroscopy (P < 0.001). Positive peritoneal cytology was noticed in 8 (6.7%) patients of which 7 were in the NS group and 1 was in the CO(2) group. Positive cytology was significantly more common among patients after hysteroscopy using NS than CO(2) (14.0% versus 1.4%, odds ratio = 11.2, 95% confidence interval = 1.3-94.5, P = 0.009). The presence of positive peritoneal cytology was not associated with age, tumor grade, tumor size, myometrial invasion, cervical involvement, or nodal metastasis. All 8 patients with positive cytology received no additional treatment and are disease free at 12 to 34 months of follow-up. CONCLUSIONS: Our data suggested that endometrial malignant cells were introduced into the peritoneal cavity during hysteroscopy and might be more likely after the use of NS rather than CO(2). This report emphasizes the need for prospective evaluation for further clarification of this hypothesis. The clinical significance of the dissemination awaits the long-term follow-up of these patients. PMID- 11855877 TI - Survival among U.S. women with invasive epithelial ovarian cancer. AB - OBJECTIVE: Invasive epithelial ovarian cancer is a highly fatal disease, diagnosed at advanced stages when survival is poor. Relatively little is known about the variation in survival across U.S. women of different race/ethnicities. To investigate this issue, we evaluated pathological characteristics and death rates due to invasive epithelial ovarian cancer in a population-based sample of patients from six racial/ethnic groups. METHODS: The analysis included 38,012 women diagnosed with primary invasive epithelial ovarian cancer between 1973 and 1997 in the Surveillance, Epidemiology and End Results Program of the National Cancer Institute. RESULTS: Filipina patients were younger at diagnosis, more likely to have localized disease, and had more mucinous cancers than whites. African-Americans were more likely than whites to be diagnosed at older ages, with distant disease and with undifferentiated/unclassified cancers. After adjusting for age at diagnosis, stage of disease at diagnosis, and cancer histology, we found that, compared to whites, death rates were significantly elevated among African-Americans and significantly reduced among Hispanics and Filipina. We also found that death rates declined significantly with time since diagnosis among women with advanced disease. CONCLUSION: The declining death rates in women with advanced disease suggest the presence of considerable prognostic heterogeneity among these women, which could reflect differences in quality of care. This issue, as well as the survival disadvantage for African American women and survival advantages for Hispanic and Filipina women, needs investigation. PMID- 11855879 TI - Preoperative CA-125 levels in patients with hereditary compared to sporadic epithelial ovarian carcinoma. AB - OBJECTIVE: The aim of this study was to determine whether a significant difference in preoperative CA-125 levels exists between patients with BRCA associated hereditary ovarian carcinoma and those with sporadic ovarian carcinoma and whether the CA-125 level predicts the probability of optimal cytoreductive surgery. METHODS: From a retrospective cohort of 189 consecutive ovarian cancer patients genotyped for BRCA mutation status, data on preoperative CA-125 levels were available for 49/88 (56%) hereditary cases and 43/101 (43%) sporadic cases. Data on the extent of surgical cytoreduction were obtained for all 92 patients with available CA-125 data. Comparison of preoperative CA-125 levels between hereditary and sporadic groups was assessed using the Kruskal-Wallis chi(2) test. Correlation of surgical cytoreduction with preoperative CA-125 level was assessed using Fisher's exact test. RESULTS: Mean preoperative CA-125 levels were not significantly different among BRCA1 (2289 U/ml), BRCA2 (2586 U/ml), and sporadic (3307 U/ml) cases (P = 0.5). For hereditary cases, optimal cytoreduction was achieved in 59% of patients with preoperative CA-125 levels of <500 U/ml and in 52% of patients with preoperative levels >500 U/ml. For sporadic cases, optimal cytoreduction was achieved in 62% of patients with CA-125 levels of <500 U/ml and in 20% of patients with levels >500 U/ml (P = 0.01). CONCLUSIONS: Preoperative CA 125 levels are not significantly different for patients with hereditary compared to sporadic ovarian carcinoma. The probability of optimal cytoreduction is independent of the preoperative CA-125 level for hereditary cases, but optimal cytoreduction is significantly less likely for sporadic cases with CA-125 levels of >500 U/ml. PMID- 11855878 TI - Time-dependent changes in factors involved in the apoptotic process in human ovarian cancer cells as a response to cisplatin. AB - OBJECTIVES: Apoptosis is believed to be a major mechanism of cisplatin-induced cell death. We investigated the kinetics of apoptosis in four human ovarian cancer cell lines treated with cisplatin to obtain insight into the role and the behavior of a variety of factors involved in this process. METHODS: The cell lines A2780, H134, and IGROV-1 (all wild-type p53) and OVCAR-3 (mutant p53) were exposed to cisplatin for 1 h and the antiproliferative effects were measured after 96 h. At various time points up to 96 h after the 1-h exposure to the individual 90% growth-inhibiting cisplatin concentrations, FACS analysis and May Grunwald Giemsa staining were carried out to determine the extent of apoptosis. At the same time points protein expression levels of p53, p21/WAF1, Bax, and Bcl 2 and the activity of caspase-3 were measured. FACS analysis was also carried out to determine changes in cell cycle distribution as a response to cisplatin. RESULTS: The four cell lines differed in sensitivity to cisplatin. A2780 was the most sensitive and IGROV-1 was the least sensitive. In contrast, IGROV-1 cells showed the highest percentage of apoptosis (30-40%), while A2780 had the lowest percentage (6-14%) (r = 0.99). The occurrence of apoptosis was not dependent on functional p53. Of interest, caspase-3 activity was in line with the percentage of apoptosis and preceded DNA fragmentation and the visualization of condensed nuclei. Wild-type p53 cells accumulated in the S phase, while OVCAR-3 arrested in the G2/M phase. The protein expression levels of p53, p21/WAF1, Bax, and Bcl-2 varied in time, but were not related to the apoptotic behavior of the cells. Upregulation of p53 was already evident before activation of caspase-3. CONCLUSIONS: Time-dependent changes in the various factors involved in the apoptotic process induced by equitoxic doses of cisplatin vary strongly among the cell lines. Caspase-3 activation plays an important role in cisplatin-induced apoptosis and this precedes morphological changes. The ability of cells to enter apoptosis, however, does not seem to predict sensitivity to cisplatin. PMID- 11855880 TI - Clinical characteristics and treatment of gestational trophoblastic tumor with vaginal metastasis. AB - OBJECTIVE: The aim of this study was to evaluate clinical manifestations, management options, and prognosis for women presenting with gestational trophoblastic tumors with vaginal metastasis. METHOD: Fifty-one patients with vaginal metastases were analyzed retrospectively between January 1985 and September 2000. Vaginal metastasis were documented by physical examination and tissue biopsy. RESULTS: The incidence of vaginal metastasis in choriocarcinoma and invasive mole was 8.6 and 4.1%, respectively. The metastatic tumors were mostly located in the anterior wall of the lower part of vagina. Eighteen patients presented with hemorrhage and rupture. All patients were treated with 5 Fu combined chemotherapy. Vaginal packing was employed to stop bleeding in 16 patients. Three of them received selective angiographic embolization. Vaginal tumors disappeared after chemotherapy. Forty-four patients with complete remission were followed up periodically without evidence of recurrence. CONCLUSIONS: Large or multiple vaginal metastases place the patients at high risk for significant hemorrhage. 5-Fu combined chemotherapy is still a reliable method for treating vaginal metastases. Angiographic embolization is emerging as a successful procedure to control the severe hemorrhage of vaginal tumors. PMID- 11855881 TI - A comparison of two prophylactic regimens for hypersensitivity reactions to paclitaxel. AB - OBJECTIVE: The aim of this study was to compare the rates of hypersensitivity reactions to paclitaxel with the conventional prophylactic regimen of two doses of oral corticosteroids and a modified regimen of a single dose of intravenous corticosteroid. METHODS: This was a retrospective historical cohort study assessing the rates of hypersensitivity reactions in patients receiving paclitaxel for ovarian or primary peritoneal carcinoma at the Hamilton Regional Cancer Centre from 1996 to 2000. Until 1998, all patients received the conventional prophylactic regimen consisting of two doses of oral dexamethasone (20 mg), 12 and 6 h prior to paclitaxel. From 1998 to the present, patients received a single dose of intravenous dexamethasone (20 mg), 30 min prior to paclitaxel. All patients also received an H(1) and H(2) blocker intravenously prior to paclitaxel administration. The analysis was corrected for potential covariates such as dose of paclitaxel and rate of infusion. The primary outcome measure was the rate of hypersensitivity reactions as defined by the National Cancer Institute of Canada-Clinical Trials Group. The Yates-corrected chi(2) test was used to compare the rates of these reactions, and a logistic regression analysis was used to determine whether any of the covariates were significant factors in these reactions. RESULTS: One hundred seven patients received the conventional corticosteroid prophylaxis prior to paclitaxel, and 110 received the single-dose intravenous corticosteroid prophylaxis. Of the 107 patients in the conventional prophylaxis group, 8 had a hypersensitivity reaction (7.5%), and only 1 of these was severe (0.9%). In contrast, of the 110 patients in the single dose IV corticosteroid group, 19 had a hypersensitivity reaction (17.3%), and 8 of these were severe (7.3%). The difference in hypersensitivity reaction rates was significant (chi(2), P = 0.047). In the logistic regression analysis, the only significant factor related to hypersensitivity reactions was the type of prophylactic steroid regimen. CONCLUSIONS: In this series, the single-dose intravenous corticosteroid prophylactic regimen appeared to be associated with a higher rate of hypersensitivity reactions to paclitaxel than the conventional two dose oral corticosteroid regimen. PMID- 11855882 TI - Laparoscopic pelvic and paraaortic lymph node dissection in the obese. AB - OBJECTIVE: The aim of this study was to determine the utility of laparoscopic pelvic and paraaortic lymph node dissection in obese women. METHODS: We performed a retrospective analysis from 1/8/96 to 1/14/01 at the University of Oklahoma Health Science Center, evaluating patients who had a Quetelet index (QI) > or =28 and had planned laparoscopic bilateral pelvic and paraaortic lymph node dissections (lnd) for their gynecologic cancer. This group was compared to a matched group of patients that had lnd done by laparotomy. Patients were identified by our institution's database and data were collected by review of their medical records. Data were collected regarding demographics, stage, histology, length of stay, and procedural information including completion rates, estimated blood loss (EBL), operating room (OR) time, lymph node count, assistant, and complications. Associations between variables were analyzed using Student t tests and chi(2) testing, Excel v9.0. RESULTS: Fifty-five patients had planned laparoscopic lnd (Group 1) and 45 patients had lnd via laparotomy (Group 2). All patients had the diagnosis of endometrial cancer. The percentage of stage I patients did not differ between groups (42/55, 71.2% versus 37/45, 82.2%, P = n.s.). Age and QI were also similar between groups, (64.6 versus 58.4, 40.0 versus 39.3, P = n.s.). Laparoscopy was completed in 35/55 (63.6%) cases. Reasons for conversion included obesity (23.6%), adhesions (1.8%), intraperitoneal cancer (5.5%), and bleeding (5.5%). QI > or =35 was associated with a decreased success rate compared to QI <35 (44.4% versus 82.1%, P = 0.004). There was no difference in successful laparoscopy when the first assistant was a fellow or a community obstetrician/gynecologist (61.0% versus 50.0%, P = n.s.). The patients in Group 1 who had laparoscopy completed had a longer OR time compared to those in Group 2 (265.3 versus 140.7 min, P < 0.0001), EBL and transfusion rates were equivalent (361.8 versus 344.2 ml, 5.6% versus 6.7%, P = n.s.), and length of stay was shorter (2.8 versus 4.5 days, P = 0.0004). Group 1 had significantly fewer postoperative fevers (5.5% versus 31.1%, P = 0.0007), fewer postoperative ileus (0% versus 13.3%, P = 0.005), and a trend for fewer wound infections (9.0% versus 22.2%, P = 0.07). CONCLUSIONS: Obesity is not a contraindication to laparoscopic pelvic and paraaortic lymph node dissection. The overall success rate was significantly higher in those patients with a QI <35. Advantages include shorter hospital stay, fewer postoperative fevers, fewer postoperative ileus, and possibly fewer wound infections. PMID- 11855883 TI - Reduced expression of early growth response-1 gene in leiomyoma as identified by mRNA differential display. AB - OBJECTIVES: Despite being the most common pelvic tumor in women, little is known about the molecular basis of uterine leiomyoma growth. The purpose of this study was to (1) identify genes important for leiomyogenesis by comparing gene expression in leiomyoma and normal myometrium and (2) examine the expression level of selected genes on a larger panel of leiomyomas. METHODS: The technique of mRNA differential display (DD) was used to perform a larger survey of gene expression in leiomyoma by comparing it to matched normal myometrium. Transcripts differentially expressed in leiomyoma were sequenced and further analyzed. Selected differentially expressed transcripts were used as probes on RNA blots of 20 pairs of leiomyomas and their matched myometria to evaluate their expression. The level of expression was analyzed by densitometry. RESULTS: From 80 differentially expressed transcripts picked by DD, 30 were sequenced and compared to GenBank data. Selected transcripts were analyzed for mRNA levels on a panel of 20 pairs of leiomyomas and matched normal myometria. One transcript that matches the early response gene EGR1 was found to be dramatically reduced (9.2 times on average) in leiomyomas. The reduction was present in 100% of the tumors. CONCLUSIONS: mRNA differential display is a valid method for finding differences in gene expression between leiomyoma and normal myometrium. The consistency and level of EGR1 mRNA reduction in leiomyoma is an indication that this gene may play a role in the origin or growth of leiomyoma. PMID- 11855884 TI - Predictors of lymphatic failure in endometrial cancer. AB - OBJECTIVE: The aim of this study was to identify determinants of lymphatic failure in patients with endometrial cancer after definitive primary treatment. METHODS: We observed 142 relapses in endometrial cancer patients who had primary surgery at our institution during the decade before 1994. We defined lymphatic failure as a relapse occurring on the pelvic sidewall (PSW), para-aortic area (PAA), or other node-bearing area (i.e., groin, axilla, supraclavicular, mediastinal). Mean follow-up was 72.8 months. RESULTS: We observed 44 instances of lymphatic failure--6 on the PSW only, 16 in the PAA only, 12 concomitantly in the PAA and on the PSW, and 10 confined in other node-bearing areas. By univariate analysis, body mass index > or = 30 kg/m(2), para-aortic lymph node biopsy, cervical stromal invasion (CSI), positive adnexa, myometrial invasion >50%, primary tumor diameter >2 cm, positive peritoneal cytology, positive lymph nodes (pelvic and/or para-aortic), radiotherapy, grade 3 tumor, nonendometrioid histology, and lymph--vascular invasion (LVI) significantly (P < or = 0.05) correlated with lymphatic failure. However, on Cox regression analysis, only LVI (P < 0.01, relative risk [RR] = 4.27), nodal involvement (P = 0.02, RR = 3.43), and CSI (P = 0.049, RR = 2.26) were independent predictors of lymphatic failure. Moreover, lymph node metastases (P = 0.01, RR = 19.82) and CSI (P = 0.050, RR = 3.57) independently predicted failure on the PSW, and only lymph node involvement (P < 0.01, RR = 10.15) predicted relapse in the PAA. CONCLUSION: LVI, positive lymph nodes, and CSI were the strongest predictors of lymphatic failure in endometrial cancer (31% of patients with at least one of the above three variables had a failure at 5 years). Patients with none of the above three factors had an extremely low (<1%) risk of lymphatic failure. PMID- 11855885 TI - Shed membrane fragment-associated markers for endometrial and ovarian cancers. AB - OBJECTIVE: The objective of this study was to assess circulating tumor-derived membrane fragments (MFs) and specific proteins associated with them as diagnostic and prognostic markers for ovarian and endometrial cancers. METHODS: The level of shed tumor-derived plasma MFs was analyzed chromatographically on high exclusion limit agarose-based gels, using sera from non-cancer-bearing female controls (n = 50), women with ovarian disease (benign, n = 43, and stages III and IV ovarian cancer, n = 62), and women with early (n = 10) and late (n = 12) stage endometrial cancers. The presence of specific proteins on MFs associated with development and progression of cancer was examined by Western immunoblot, while the association of proteolytic enzymes with MFs was analyzed by zymography. RESULTS: Shed MFs were demonstrated in the circulation of women with both ovarian (4.12 plus minus 1.46 mg/ml) and endometrial cancers (2.53 +/- 0.34 mg/ml in women with stage I and 4.51 +/- 1.33 mg/ml with late stage); however, they were not demonstrated in control sera or in sera from women with benign disease. In endometrial cancer, the level of MFs correlated with the tumor's progression. Specific proteins, including MMP-2, MMP-9, and Fas ligand (FasL), were present on MFs from both endometrial and ovarian cancer sera. However, FasL (3.2-fold) and MMP-2 and -9 (5.9x and 7.5x, respectively) levels were significantly elevated on MFs from late stage cancer. CONCLUSION: This study demonstrates the unique elevation of circulating MFs in ovarian and endometrial cancer patients. The levels of specific MF-associated proteins differentiate between early and late stage endometrial cancers and benign versus malignant ovarian disease. PMID- 11855886 TI - Lymphatic mapping of sentinel nodes in early vulvar cancer. AB - OBJECTIVE: The aim of the study was to determine the diagnostic accuracy and feasibility of sentinel lymph node (SLN) detection using a gamma probe in patients suffering from vulvar cancer. METHODS: From May 1998 to November 2000, 26 patients with early vulvar cancer, planned for local wide excision or vulvectomy including groin dissection, were eligible for the study. Two to 3 h before the planned procedure we injected technetium(99) m-labeled microcolloid intradermally at four locations around the tumor. Dynamic and static images were recorded using a gamma camera. SLN locations were marked on the overlying skin. In the operating theater SLNs were identified at the beginning of the procedure using a handheld gamma-detection probe. After resection of suspected SLNs a standard unilateral or bilateral groin dissection was performed, subsequently followed by local wide excision or, if indicated, radical vulvectomy. Sentinel node detection using technetium(99) m-labeled microcolloid was compared with final histopathological and immunohistochemical results. RESULTS: Scintigraphy showed focal uptake in all 26 patients. Intraoperatively we detected all sentinel nodes by handheld gamma probe. In 20 patients, one sentinel node was identified unilaterally, while in 6 patients two or more nodes were identified bilaterally. Histologically positive SLNs were found in 9 patients. In our preliminary series we did not find any false-negative SLN. CONCLUSION: Identification of sentinel nodes in vulvar cancer is feasible with preoperatively administered technetium(99)m-labeled microcolloid. We confirm the results of previous studies and improve the evidence that the SLN procedure could be implemented in future therapy concepts. PMID- 11855887 TI - Papillary serous adenocarcinoma of the ovary diagnosed after malignant pericardial tamponade and embolic stroke. AB - BACKGROUND: Epithelial carcinomas of the ovary are predominantly an intraperitoneal disease. Reports of epithelial ovarian carcinomas metastatic to the pericardium are rare. CASE: A 43-year-old woman was admitted with symptoms of a pericardial tamponade, as well as an embolic cerebrovascular accident, and transferred to the ICU where a pericardiocentesis was performed. Cytology revealed malignant cells in the pericardial fluid. CT scan of the abdomen and pelvis revealed bilateral pelvic masses. A laparotomy revealed a papillary serous adenocarcinoma of ovarian primary and an infarcted spleen with capsular tumor metastases. The malignant cells in the pericardial fluid were consistent with the ovarian primary. CONCLUSION: Ovarian cancer metastasis to the heart and pericardium presented an aggressive variant of tumor spread with significant morbidity and subsequent mortality. PMID- 11855888 TI - Iliac atherosclerotic occlusive disease complicating radiation therapy for cervix cancer: a case series. AB - BACKGROUND: Diagnosis and management of atherosclerotic occlusive disease complicating radiation therapy for squamous carcinoma of the cervix (SCC) are not well known. CASES: A series of four patients who underwent medical or surgical treatment of radiation-induced artery disease were identified. Chronic right iliac artery occlusion was seen in one patient 7 years after external irradiation for SCC and was treated by a femorofemoral bypass. Acute arterial occlusion was seen in three patients. One patient developed right foot ischemia 2 years after external irradiation for SCC which was treated by a right axillofemoral bypass. One patient developed a left toe necrosis 2 years after external irradiation for a recurrence of SCC which was treated by a percutaneous artery dilatation. One patient developed, 3 years after external irradiation for SCC, a subacute ischemia of the left leg on the first postoperative day after a rectal resection. It was treated by vasodilators, calcium inhibitors, and anticoagulants. CONCLUSIONS: Diagnosis of radiation-induced atherosclerotic occlusive disease complicating radiation therapy for SCC is easy but treatment is difficult. Long term results of arterial dilatation are limited and extraanatomic bypass with allograft is the treatment of choice whenever feasible. However, because of poor long-term results, surgery should be attempted only in the case of severe ischemia. PMID- 11855890 TI - Tumor chemoconversion following surgery, chemotherapy, and normalization of serum tumor markers in a woman with a mixed type germ cell ovarian tumor. AB - BACKGROUND: Malignant germ cell tumors of the ovary are often curative after conservative surgery and adjuvant chemotherapy. Persistent tumors despite normalization of serum tumor markers may represent retroconversion to benign masses, but this is rare in ovarian tumors without teratoma elements. The management in such cases has not been defined. CASE: A young woman with a stage IIIC mixed germ cell ovarian tumor containing endodermal and dysgerminoma elements and elevated serum tumor markers underwent conservative surgery followed by chemotherapy. Residual tumor persisted on CT despite the normalization of serum tumor markers. The residual tissue was resected and contained benign tissue. CONCLUSIONS: In cases where masses persist and serum tumor markers normalize, attaining a histological diagnosis, and not chemotherapy, should be considered. PMID- 11855889 TI - Simultaneous sertoli cell tumor and adenocarcinoma of the tunica vaginalis testis in a patient with testicular feminization. AB - BACKGROUND: The association of testicular feminization with late diagnosis in a patient with a large Sertoli cell tumor and a metastasizing adenocarcinoma of the tunica vaginalis testis is unusual. CASE: Testicular feminization was diagnosed in a 72-year-old patient, who was admitted with a large lower abdominal mass. Histologically, we found a well-differentiated Sertoli cell tumor and an adenocarcinoma of the tunica vaginalis testis with metastases in the sigmoid colon, rectum, and omentum. Explorative laparotomy revealed a large pelvic tumor mass and extensive peritoneal carcinosis. After debulking surgery to optimal residual disease and four courses of chemotherapy (cisplatin and etoposide), there was no evidence of disease (clinically) for 24 months before an intraabdominal and inguinal relapse occurred. Due to the unwillingness of the patient to receive salvage chemotherapy or palliative abdominal surgery, the disease progressed rapidly and she died 27 months after the initial operation. CONCLUSION: This is the first reported case of an advanced carcinoma of the tunica vaginalis testis occurring simultaneously with a large Sertoli cell tumor in a patient with testicular feminization. Surgical debulking and platinum-based chemotherapy rendered the patient clinically free of disease for 2 years. PMID- 11855891 TI - Ovarian mucinous cystadenocarcinoma with virilization. AB - BACKGROUND: Ovarian neoplasms, other than sex cord--stromal tumors, are rare causes of hyperandrogenism. Only two cases of primary mucinous carcinomas associated with virilization have been reported. CASE: A 50-year-old female was referred to our clinic with a large pelvic mass. On examination she had significant facial hirsutism, clitoromegaly, and male pattern pubic hair growth. Serum levels of testosterone and dihydroepiandrosterone sulfate were elevated. A 30-cm, multilocular, solid and cystic, left ovarian mass was resected. Histology revealed moderately to poorly differentiated mucinous cystadenocarcinoma. The ovarian stroma contained florid proliferation of luteinized cells. The right ovary showed cortical stromal hyperplasia. Abnormal hormone values normalized 10 days postoperatively. CONCLUSION: We report a rare case of mucinous cystadenocarcinoma with virilization, review the literature, and discuss the mechanisms of hormone production by these tumors. PMID- 11855892 TI - Laparoscopic management of urolithiasis in a continent urostomy. AB - BACKGROUND: Urinary calculi formation is an infrequent but challenging late complication of continent urostomy. Percutaneous endoscopic management of two patients with symptomatic pouch urolithiasis is described. CASES: A 48-year-old woman with a history of cervical squamous cell carcinoma (SCC) treated with primary chemoradiotherapy presented with recurrent pain and urinary tract infections (pouchitis). She had undergone continent urostomy 4 years previously for management of a vesicovaginal fistula. A 59-year-old woman following exenteration with continent diversion for recurrent cervix SCC had a 6-month history of refractory pouchitis. In both women, multiple pouch urinary calculi were identified. Their continent reservoirs were, in part, created using titanium staplers. Percutaneous CO(2) endoscopy afforded identification and removal of their stones and staples without complication. CONCLUSION: Successful treatment of continent urostomy urolithiasis is accomplished by percutaneous endoscopy. PMID- 11855893 TI - Incidence of positive peritoneal cytology in low-risk endometrial cancer treated by laparoscopically assisted vaginal hysterectomy. PMID- 11855895 TI - Elevated platelet counts in patients with breast cancer. PMID- 11855896 TI - Implication of a reflected image illustration for pelvic lymphadenectomy on uterine cervical cancer with situs inversus totalis. PMID- 11855897 TI - Social suppression in nonreproductive female Damaraland mole-rats, Cryptomys damarensis: no apparent role for endogenous opioid peptides. AB - The role of endogenous opioid peptides (EOPs) on LH secretion was examined to investigate the neuronal mechanisms responsible for the inhibition of GnRH and the resultant infertility in nonreproductive female Damaraland mole-rats, Cryptomys damarensis. The endorphin antagonist naloxone was administered to five groups of females to determine its effect on plasma LH levels: Grouping was determined by social status, social environment, and whether the females were ovariectomized. A single injection of naloxone had no significant effect on LH secretion in either intact or hystero-ovariectomized females. Multiple injections with naloxone failed to affect basal LH concentrations but did result in a decrease in GnRH-stimulated LH secretion in ovariectomized nonreproductive and reproductive females. A significant response to a single naloxone injection following GnRH priming was obtained in both nonreproductive females and in nonreproductive females housed in the absence of the reproductive pair. These results suggest EOPs play a role in sexual function but that socially induced infertility is unlikely to be mediated through the EOP system. PMID- 11855898 TI - MPOA cytotoxic lesions and maternal behavior in the rat: effects of midpubertal lesions on maternal behavior and the role of ovarian hormones in maturation of MPOA control of maternal behavior. AB - Small neurotoxin lesions in the medial preoptic area (MPOA) block maternal behavior (MB) in adults but large lesions are required to produce the same effect in juvenile rats (23-27 days of age). To study the maturation of MPOA control of MB, in Experiment I, we compared the effects of small versus large neurotoxin MPOA lesions at midpuberty (38 days of age) on MB. Midpubertal females with large MPOA lesions showed severe impairment in MB affecting retrieving, crouching, and nest building, but 85% of females with small MPOA lesions exhibited all components of MB and performed like control females without MPOA lesions. To study the role of ovarian hormones during puberty on the maturation of MPOA mediation of MB (Experiment IIA), females were ovariectomized either before or after puberty and small MPOA cytotoxic lesions were made at 53 days of age. At 60 days of age both groups showed similar deficits in MB which indicated that the maturation of the MPOA mediation of MB is not dependent on pubertal ovarian hormones. In Experiment IIB, we administered estradiol benzoate (sc) and this overcame the deficit in MB after small MPOA lesions in females that had been deprived of estrogen for shorter periods (30 days) but had not been deprived for longer periods (60 days). In addition, ovary-intact females with circulating estrogen and small lesions in the MPOA at 53 days of age did not show deficits in MB. PMID- 11855899 TI - Cognitive and behavioral characteristics of turner syndrome: exploring a role for ovarian hormones in female sexual differentiation. AB - To better understand factors contributing to behavioral development, we studied patients with Turner syndrome (TS), a disorder typically marked by prenatal onset of ovarian dysfunction. We compared girls and women (ages 12 and up) with TS (n = 21) to matched controls (n = 21) in cognitive and motor skills, as well as sex typed personality characteristics and activity preferences. Measures were categorized (based on prior studies) as showing an average male advantage (male superior measures), female advantage (female-superior measures), or no sex difference (sex-neutral measures). It was hypothesized that, if gonadal function contributes to behavioral development, effects of this deficiency would be more prominent on sexually differentiated than sex-neutral measures and thus that patient-control differences would be most marked for measures that show sex differences. Our findings indicated that TS patients and controls differed more on cognitive and motor domains that show sex differences than on sex-neutral domains. Patients also had more "undifferentiated" personalities and showed reduced sex-typed interests and activities. Differing experiences, as indexed by interests and activities, did not explain the observed cognitive and motor differences. These results are consistent with a role for ovarian hormones acting on the brain to influence cognitive and behavioral development, although they do not rule out other possible interpretations. PMID- 11855900 TI - Natural breeding conditions and artificial increases in testosterone have opposite effects on the brains of adult male songbirds: a meta-analysis. AB - A meta-analysis of the literature shows that in adult male songbirds, brain mass, telencephalon volume and n. rotundus (a thalamic visual nucleus) volume increase from the nonbreeding season (low testosterone) to the breeding season (higher testosterone). These effects can at least partially be mimicked by photoperiod manipulations in captivity. In contrast, an artificial testosterone (T) titer increase by chronic implants yields the opposite results: telencephalon, n. rotundus, and n. pretectalis volumes are lower in T-treated animals than in controls. These results suggest that artificial testosterone manipulations do not necessarily mimic the effects of natural variations in hormone levels and that results from experiments using T implants to mimic natural hormonal effects should be interpreted with caution. PMID- 11855901 TI - Oxytocin maintains as well as initiates female sexual behavior: effects of a highly selective oxytocin antagonist. AB - In previous studies, central administration of the oxytocin (OT) antagonist d(CH2)5[Tyr(Me)2, Thr4, Tyr-NH(9)2]OVT (OTA1) blocked receptive and proceptive components of female sexual behavior (FSB) and increased male-directed agonistic behavior when given before progesterone (P) treatment in estradiol-primed female rats but not when given shortly before behavioral testing 4-6 h after P. Because the considerable V(1a) antagonist potency of OTA1 may have contributed to these results, we tested the effects of the far more selective OT antagonist desGly NH2, d(CH2)5[d-Tyr2, Thr4]OVT (OTA2). In ovariectomized, estradiol benzoate primed (1 microg x 2 days sc) rats, icv infusion of OTA2 (1 microg) prior to P injection (250 microg sc) significantly suppressed lordosis and hops and darts and trended toward significantly increasing male-directed kicks during testing at 4 and 6 h. Infusion of OTA2 3 h and 40 min after P did not alter behavior at 4 and 6 h after P but significantly decreased lordosis as well as hops and darts and increased male-directed kicks 8-12 h after P. These results provide further evidence that central OT receptor activation shortly after P treatment contributes to the subsequent onset and early expression of FSB and demonstrate, for the first time, that OT receptor activation at later time points also contributes to maintaining FSB. The FSB-stimulating effect of central OT appears to persist for several hours. PMID- 11855902 TI - Effects of testosterone on song, aggression, and nestling feeding behavior in male great tits, Parus major. AB - As outlined in the trade-off hypothesis of testosterone (T) secretion, fluctuations in T during the breeding season might reflect how males allocate their time and energy to competitive behaviors for mates and territories, associated with high T levels, and parental activities, associated with low T levels. In the present study, great tit, Parus major, males were implanted with T filled or empty silastic capsules at the start of the breeding season and the behavior of these two male categories was compared during the entire breeding season. As a measure of competitive behavior we looked at song behavior and territorial responsiveness to a male decoy, during the three main stages of the breeding period (the egg-laying, incubation, and nestling stages). As a measure of parental care we looked at feeding behavior during the nestling stage. Our results only partly supported the trade-off hypothesis. T implants increased plasma androgen levels and enhanced spontaneous song activity and the production of aggressive vocalizations in response to a decoy. However, our results suggest that the degree of physical aggression might be less than fully coupled with T. First, approach to the decoy was not affected by the treatment. Second, although T levels are known to vary from high during egg laying to low while feeding young, control and T-treated males spent similar amounts of time close to the decoy in the three breeding stages. Our results thus suggest that vocal and physical aggression might be regulated differently in the great tit. Furthermore, in contrast with most other studies on temperate bird species but in agreement with a previous study on the great tit, T treatment did not affect male feeding rates. As the dose of T we used was lower than that typically used in other studies, we cannot at present completely exclude the possibility that the latter result reflects this lower dose of T rather than the species used. PMID- 11855903 TI - Hormonal and body size correlates of electrocommunication behavior during dyadic interactions in a weakly electric fish, Apteronotus leptorhynchus. AB - Brown ghost knife fish, Apteronotus leptorhynchus, produce sexually dimorphic, androgen-sensitive electrocommunication signals termed chirps. The androgen regulation of chirping has been studied previously by administering exogenous androgens to females and measuring the chirping response to artificial electrical signals. The present study examined the production of chirps during dyadic interactions of fish and correlated chirp rate with endogenous levels of one particular androgen, 11-ketotestosterone (11KT). Eight males and four females were exposed to short-term (5-min) interactions in both same-sex and opposite-sex dyads. Twenty-four hours after all behavioral tests, fish were bled for determination of plasma 11KT levels. Males and females differed in both their production of chirps and their ability to elicit chirps from other fish: males chirped about 20-30 times more often than females and elicited 2-4 times as many chirps as females. Among males, chirp rate was correlated positively with plasma 11KT, electric organ discharge frequency, and body size. Combined with results from experimental manipulation of androgen levels, these results support the hypothesis that endogenous 11KT levels influence electrocommunication behavior during interactions between two male fish. PMID- 11855904 TI - Implicit power motivation predicts men's testosterone changes and implicit learning in a contest situation. AB - This study tested the hypothesis that implicit power motivation moderates men's testosterone responses to victory or defeat in a contest situation. It also explored to what extent postvictory testosterone increases are associated with enhanced implicit learning of behavior instrumental for winning a contest. Salivary testosterone levels were assessed in 66 male adults several times before and after a contest whose outcome (winning or losing against a competitor on an implicit learning task) was varied experimentally. Among participants low in activity inhibition, a measure of impulse control, the power motive was a significant positive predictor of testosterone increases (15 min postcontest; r = 0.71, P = 0.01) and implicit learning (r = 0.68, P < 0.05) after a victory, whereas it was a significant negative predictor of implicit learning (r = -0.58, P = 0.01) but not of testosterone increases (r = -0.08, ns) after a defeat. Moreover, among participants low in activity inhibition testosterone increases were associated with enhanced implicit learning (r = 0.38, P < 0.05) and there was statistical evidence that in winners testosterone increases mediated the effect of power motivation on implicit learning. Participants high in activity inhibition did not display this pattern of results. PMID- 11855905 TI - Dehydroepiandrosterone (DHEA) increases territorial song and the size of an associated brain region in a male songbird. AB - In many species, male territorial aggression is tightly coupled with gonadal secretion of testosterone (T). In contrast, in song sparrows (Melospiza melodia morphna), males are highly aggressive during the breeding (spring) and nonbreeding (autumn and early winter) seasons, but not during molt (late summer). In aggressive nonbreeding song sparrows, plasma T levels are basal (< or = 0.10 ng/ml), and castration has no effect on aggression. However, aromatase inhibitors reduce nonbreeding aggression, indicating a role for estrogen in wintering males. In the nonbreeding season, the substrate for brain aromatase is unclear, because plasma T and androstenedione levels are basal. Aromatizable androgen may be derived from plasma dehydroepiandrosterone (DHEA), an androgen precursor. DHEA circulates at elevated levels in wintering males (approximately 0.8 ng/ml) and might be locally converted to T in the brain. Moreover, plasma DHEA is reduced during molt, as is aggression. Here, we experimentally increased DHEA in wild nonbreeding male song sparrows and examined territorial behaviors (e.g., singing) and discrete neural regions controlling the production of song. A physiological dose of DHEA for 15 days increased singing in response to simulated territorial intrusions. In addition, DHEA treatment increased the volume of a telencephalic brain region (the HVc) controlling song, indicating that DHEA can have large scale neuroanatomical effects in adult animals. The DHEA treatment also caused a slight increase in plasma T. Exogenous DHEA may have been metabolized to sex steroids within the brain to exert these behavioral and neural effects, and it is also possible that peripheral metabolism contributed to these effects. These are the first results to suggest that exogenous DHEA increases male-male aggression and the size of an entire brain region in adults. The data are consistent with the hypothesis that DHEA regulates territorial behavior, especially in the nonbreeding season, when plasma T is basal. PMID- 11855906 TI - Sex difference in attraction thresholds for volatile odors from male and estrous female mouse urine. AB - Volatile urinary odors from opposite sex conspecifics contribute to mate recognition in numerous mammalian species, including mice. We used a simple habituation/dishabituation testing procedure to ask whether the capacity to detect and investigate decreasing concentrations of volatile urinary odors is sexually differentiated in mice. Beginning 2 months after gonadectomy and in the absence of any sex steroid treatment, adult, sexually naive male and female CBA x C57Bl/6 F1 hybrid mice received two series of daily tests that involved the presentation of different dilutions of urine from C57Bl/6 males followed by urine from estrous females. Each test session began with three consecutive presentations of deionized water (10 microl on filter paper for 2 min, behind a mesh barrier which prevented direct physical access, in the home cage at 1-min intervals) followed by three presentations of one of five different dilutions of urine (a different dilution on each test day). Males and females showed equivalent, significant habituation/dishabituation responses (low investigation times for successive water presentations; increased investigation of the first urine stimulus, followed by a decline in successive urine investigation times) to both male and female urine/water dilutions of 1:1, 1:10, and 1:20. However, only female mice responded reliably to 1:40 and 1:80 dilutions of both types of urine, pointing to a sex dimorphism in the detection and/or processing of biologically relevant, volatile urinary odors by the main olfactory system. PMID- 11855907 TI - Resveratrol: phytoestrogen effects on reproductive physiology and behavior in female rats. AB - Resveratrol is a phytoestrogen naturally found in grapes and is a major constituent of wine thought to exert both cardioprotective and chemopreventive activities. Recent studies show that this bioflavonoid binds to and activates gene transcription via the estrogen receptor (ER) subtypes ERalpha and ERbeta. Previous studies have focused primarily on the in vitro effects of resveratrol (RES) in estrogen-sensitive tissues or in carcinogenic cell lines, while frequently neglecting to document its potential effects in animal models with intact neuroendocrine systems. However, the present studies were designed to systematically characterize the in vivo effects of RES on reproductive physiology and behavior in adult female rats. In gonadally intact females, RES consumption reduced body weight, disrupted estrous cyclicity, and induced ovarian hypertrophy. However, in ovariectomized females RES (10-1000 microg) injections did not appear to mimic 17 beta-estradiol benzoate (EB)-induced behavioral responses and had no lasting effects on subsequent estrogen sensitivity or sociosexual behavior. The present studies support recent in vitro findings that RES differs from other phytoestrogens by acting as a possible mixed agonist/antagonist, depending on the availability of specific ER isoforms localized in the reproductive tract and brain of the female rat. PMID- 11855908 TI - Postparturitional testosterone surge in male offspring of rats stressed and/or fed ethanol during late pregnancy. AB - Male offspring of rats exposed to restraint stress and/or alcohol during late pregnancy show aberrant patterns of sexual behavior masculinization and defeminization that vary as a function of treatment. The impact of these treatments on the postparturitional testosterone (T) surge that contributes to sexual behavior differentiation was investigated. Plasma T was measured using radioimmunoassay in individual males sampled on day 21 of gestation within 10 min of cesarean delivery or 1, 2, or 4 h thereafter. Neonatal T in the group exposed only to stress did not differ from that in the control group. T was lower than control levels at birth in both alcohol groups. The magnitude of the T surge that occurred during the first hour of birth in the control group was diminished by 50% in both alcohol groups, whose T pattern was very similar. There was no common alteration in postparturitional T associated with the increased lordotic behavior potential that males in all three treatment groups typically share, nor were there idiosyncratic endocrine abnormalities linked to the very different male copulatory pattern each exhibits. Exposure to an abnormal T milieu during fetal as well as neonatal ontogeny may underlie the etiology of the different sexual behavior patterns exhibited by males exposed to stress and/or alcohol. Possible unique effects each treatment exerts on perinatal plasma T and it's aromatization to estradiol in hypothalamic targets are discussed. PMID- 11855909 TI - Oral estrogen masculinizes female zebra finch song system. AB - It is well established that parenteral treatment of female zebra finch chicks with estradiol masculinizes their song control nuclei and that as adults they are capable of song. Concern over the widespread use of putative environmental estrogens caused us to ask whether oral exposure to estrogens (a natural route of exposure) could produce similar effects. We dosed chicks orally with estradiol benzoate (EB; 1, 10, 100, and 1000 nmol/g of body mass per day, days 5-11 posthatch), the non-ionic surfactant octylphenol (100 and 1000 nmol/g), or the pesticides methoxychlor (100 and 1000 nmol/g) and dicofol (100 nmol/g) and measured their song control nuclei as adults. EB treatment produced increases in song nuclei comparable to that induced by parenteral administration of estrogens. This is the first study of which we are aware to use an oral route of administration, which simulates the natural process of parent birds feeding their nestlings. We conclude that oral exposure to estradiol alters song control nuclei and we report in a related paper (Millam et al., 2001) that such exposure severely disrupts reproductive performance. Although we detected no influence of xenobiotics on induction of song control nuclei the possibility remains that oral exposure to xenoestrogens in high enough doses could affect development. PMID- 11855911 TI - On the mechanism of colonic motility: the electric activation theory of colonic contraction. AB - PURPOSE: A previous study has demonstrated that the longitudinal muscle possesses electric activity, while the circular does not (A. Shafik and A. A. Shafik, 2000, Front. Biosci. 5, b5). In the current study, we investigated the mechanism of action of the two colonic muscle coats in the motility of the gut. METHODS: Fourteen patients (43.8 years, 10 men) with left colon or rectal cancer were scheduled to have transverse colostomy as a part of their operation. The electric activity of the ascending colon was recorded by three electrodes applied to each of the circular and the longitudinal (taenia coli) muscle coats. Simultaneously, the colonic pressure was recorded before and after colonic distension. The test was repeated after longitudinal muscle anesthetization. RESULTS: Electric waves in the form of pacesetter (PPs) and action (APs) potentials were recorded from the longitudinal but not the circular muscle fibers. APs were associated with an intracolonic pressure rise. Colonic distension produced significant increase in the PPs and APs recorded from the longitudinal muscle with appearance of similar electric activity from the circular muscle. Electric activity and colonic pressure increased upon increase in the colonic distension until the balloon in the proximal part of the ascending colon moved to the transverse colon in one mass contraction. Ten minutes after longitudinal muscle anesthetization, no electric activity was recorded from the longitudinal and circular muscles upon colonic distension. CONCLUSIONS: The electric waves appear to be transmitted from the longitudinal to the circular muscle upon colonic distension. The giant migrating contractions of the colon that move the food bolus from the cecum to the transverse colon are suggested to be a function of the longitudinal muscle electric activation with gut lumen modulation by the circular fibers. PMID- 11855910 TI - Effect of creatine monohydrate on cardiac function in a rat model of endotoxemia. AB - BACKGROUND: Reports have attributed cardiac failure during acute models of endotoxemia to a lack of high-energy phosphates. This study was undertaken to investigate whether creatine (Cr) administered during perfusion could enhance myocardial protection and improve recovery of cardiac function in a rat model of endotoxemia. METHODS: Acute endotoxemia was induced in rats by a bolus injection of Escherichia coli endotoxin (LPS: 4 mg/kg, ip) while control rats were injected with an equal volume of 0.9% normal saline. To assess the adequacy of energy metabolism, freeze-clamped hearts were obtained from animals to study the concentrations of endogenous ATP, phosphocreatine (PCr), inorganic phosphate (P(i)), and intracellular pH by (31)P-cryomagnetic resonance spectroscopy. In a separate experiment, isolated hearts were perfused via a Langendorff column with Krebs-Henseleit buffer containing different concentrations of creatine monohydrate (1, 3, or 10 mM). Cardiac performance was evaluated via a paced (300 bpm) isovolumetric balloon preparation. Measurements of cardiac function including left ventricular developed pressure (LVDP), the maximum rates of ventricular pressure rise (LV +dP/dt) and fall (LV -dP/dt), and coronary flow were made for both LPS and saline-treated animals. RESULTS: High-energy phosphate ratios of PCr/ATP and PCr/P(i) in hearts declined significantly at 4 h after endotoxin treatment. As anticipated, LVDP and LV +dP/dt(max) at a given preload and heart rate were significantly (P < 0.05) lower at 4 h when measured at the same time point. The functional recovery of these parameters was not improved by the addition of creatine monohydrate to the perfusion buffer. Creatine produced a significant (P < 0.05) negative inotropic effect in hearts from saline-treated animals. The LVDP was reduced by 30% at the lowest concentration and by 50% at the highest concentration of creatine monohydrate. Furthermore, creatine significantly (P < 0.05) reduced LV -dP/dt(max) in both saline and LPS-treated rats. These data demonstrate that exogenous creatine does not contribute to myocardial preservation in endotoxemia. CONCLUSIONS: Energy stores in the rat heart decline early in endotoxemia accompanied by reduced myocardial performance, suggesting that the ability of the heart to perform mechanical work is impaired. Cardiac dysfunction in an acute model of endotoxemia was not improved with exogenous creatine during perfusion. Creatine's effects were primarily lusitropic by delaying the onset of myocardial relaxation in all hearts. The deleterious effects of exogenous creatine monohydrate in normal hearts should be examined in future experimental studies. PMID- 11855912 TI - l-[1-(13)C]Alanine is a useful substance for the evaluation of liver function. AB - BACKGROUND: Using a rat model of hepatectomy, we investigated whether the severity of hepatopathy could be quantitatively measured from changes in expiratory (13)CO(2) levels after intravenous administration of l-[1 (13)C]alanine. MATERIALS AND METHODS: Under nembutal anesthesia, 20 mg/kg l-[1 (13)C]alanine was administered to rats via the femoral vein, and expiratory (13)CO(2) levels were measured for 15 min. Then, 30, 70, or 90% hepatectomy was performed. In the control group, simple laparotomy was performed. A breath test was conducted 20 min after laparotomy. We examined the correlation of total (13)CO(2) output (S) or single point (13)CO(2) level (SP) every 1 min for 15 min with liver weight/body weight (LW/BW) (%). RESULTS: In the control group, the breath test graph showed a specific peak level about 3 min after administration, but in all groups undergoing hepatectomy, it did not show any peak level during measurement. The correlation coefficient between S(12--15) after l-[1 (13)C]alanine administration and LW/BW was 0.902 (P < 0.0001). The correlation coefficient between SP(7) after l-[1-(13)C]alanine administration and LW/BW was highest, 0.908 (P < 0.0001). The severity of hepatopathy could also be evaluated, with significant differences in S(12-14) compared to control when the volume of resected liver was 30% or greater, but there was no significant difference between the groups undergoing 70 and 90% hepatectomy. However, the severity of hepatopathy could be evaluated, with significant differences in S(15) and SP(7) in all comparisons between groups. CONCLUSION: In the breath test with intravenously administered l-[1-(13)C]alanine, the severity of hepatopathy could be quantitatively evaluated in a short period by measuring S(15) and SP(7). PMID- 11855913 TI - Autologous skin transplantation: comparison of minced skin to other techniques. AB - BACKGROUND: Skin grafting may be necessary to close nonhealing skin wounds. This report describes a fast and minimally invasive method to produce minced skin suitable for transplantation to skin wounds. The technique was evaluated in an established porcine skin wound healing model and was compared to split-thickness skin grafts and suspensions of cultured and noncultured keratinocytes. MATERIALS AND METHODS: The study included 90 wounds on 3 pigs. Fluid-treated full-thickness skin wounds were grafted with minced skin, split-thickness skin grafts, noncultured keratinocytes, or cultured keratinocytes. Controls received either fluid or dry treatment. The wound healing process was analyzed in histologies collected at Days 8 to 43 postwounding. Wound contraction was quantified by photoplanimetry. RESULTS: Wounds transplanted with minced skin and keratinocyte suspension contained several colonies of keratinocytes in the newly formed granulation tissue. During the healing phase, the colonies progressed upward and reepithelialization was accelerated. Minced skin and split-thickness skin grafts reduced contraction as compared to keratinocyte suspensions and saline controls. Granulation tissue formation was also reduced in split-thickness skin-grafted wounds. CONCLUSIONS: Minced skin grafting accelerates reepithelialization of fluid-treated skin wounds. The technique is faster and less expensive than split thickness skin grafting and keratinocyte suspension transplantation. Minced skin grafting may have implications for the treatment of chronic wounds. PMID- 11855914 TI - Adenovirus mediated gene transfer of antiapoptotic protein in hepatic ischemia reperfusion injury: the paradoxical effect of Bcl-2 expression in the reperfused liver. AB - BACKGROUND: Although Bcl-2 is well known to have antiapoptotic activities in vitro and in vivo, the role of Bcl-2 remains controversial. In the present study, we evaluated whether the adenovirus mediated gene transfer of hBcl-2 could exert an antiapoptotic effect in a rat model of hepatic ischemia-reperfusion (I/R) injury. MATERIALS AND METHODS: Each 6 x 10(9) plaque forming unit adenovirus vector encoding LacZ (AxCAilacZ) or hBcl-2 (AxCAhbcl2) was intravenously administered 48 h before I/R injury, in groups 1 and 2, respectively. In group 3, 1 ml of normal saline was injected instead of the virus vectors. Hepatic I/R injury was induced by the temporal occlusion of all hepatic influent vessels for 30 min under a portosystemic shunt. The animals were sacrificed at 6 h, 1, 3, and 14 days after reperfusion (each n = 12 in groups 1 and 2, and n = 8 in group 3). The expressions of hBcl-2 and Bax were evaluated at both the mRNA level by reverse transcription polymerase chain reaction and the protein level by immunohistochemistry. To assess the hepatocyte and sinusoidal endothelial cell damage after I/R injury, the serum asparate aminotransferase (AST), alanine aminotransferase, and hyaluronic acid were all evaluated. The number of apoptotic cells was evaluated by terminal deoxynucleotidyl transferase-mediated dUTP biotin nick end-labeling (TUNEL). RESULTS: To evaluate the antiapoptotic activities of the hBcl-2 sequence encoded into AxCAhbcl2, rat hepatocarcinoma cells were transfected with AxCAhbcl2 (10(3) moi) or AxCAilacZ (10(3) moi) and then challenged with TGF-beta1 protein (5 ng/ml) to induce apoptosis. Apoptotic cells were counted by TUNEL staining in about 2500 cells, and it was found that adenovirus mediated gene transfer of hBcl-2 significantly protected rat hepatocarcinoma cells from TGF-beta1 induced apoptosis (14.2 +/- 1.2%) in comparison to those of LacZ (21.9 +/- 1.4%). In the reperfused liver in vivo, the mRNA expression of hBcl-2 was detected only in the hBcl-2 transfected group 2. In group 2, a strong degree of Bcl-2 immunoreactivity was recognized as early as 6 h after reperfusion, while it was not recognized in groups 1 and 3 at 6 h after reperfusion. The AST levels were significantly higher in group 2 (AST: 356 +/- 100.1 IU/L) than those in group 1 (AST: 102.7 +/- 15 IU/L) at 1 day after reperfusion (P < 0.05). The number of TUNEL positive cells was significantly higher in group 2 than in groups 1 and 3 at 1 day after reperfusion. CONCLUSIONS: These results indicated that an overexpression of antiapoptotic protein Bcl-2 paradoxically exerted a proapoptotic effect in the reperfused liver. The in vivo role of Bcl-2 should thus be carefully evaluated, depending on the levels of expression and the target organ. PMID- 11855915 TI - Apoptosis and CD8 and CD54 cell expression in rat small bowel transplantation. AB - The importance of activated CD8 cells expressing IL-2R in small bowel and other organ rejection has been reported. Some authors even consider that a positive correlation might be demonstrated between the number of apoptotic enterocytes and the degree of graft rejection. In addition, moderate to intense activation of endothelial molecules in small bowel allograft in rats has been reported in chronic rejection. The aim of the present paper is to ascertain, in a heterotopic small bowel transplantation (HSBT) in rats, whether CD3, CD4, CD8, and CD54 cell expression in the allograft infiltrates shows some relationship with allograft enterocyte apoptosis when rejection is present. Wistar Furth male rats were allotted to two groups: group A was the control group without transplantation; group B received a heterotopic small bowel allograft from Fisher rats and an im dose of FK506 (0.25 mg/kg/day). A significant increase of CD8, CD54 cell receptor expression, and apoptosis in the group undergoing HSBT showed rejection. No significant differences have been observed in the variables under study between the control and HSBT without rejection groups or in CD3 and CD4 among the three groups. We observed a significant correlation between apoptosis and rejection, between CD8 and CD54 with apoptosis and with rejection, and between CD8 and CD54. This indicates that the activation of endothelial molecules and cells may play an important role in established HSBT chronic rejection. We consider that this study may contribute to the knowledge of small bowel allograft chronic rejection and its immunomodulation. PMID- 11855916 TI - The influence of platelet-smooth muscle cell interaction on the oxidative modification of low-density lipoprotein. AB - OBJECTIVE: In view of the recognized association between thrombosis and atherosclerosis, it is hypothesized that exposure of arterial smooth muscle cells (SMC) to thrombogenic agents such as platelets and thrombin will alter the oxidation of low-density lipoprotein (LDL) and that this effect may be diminished by thrombin inhibition. METHODS: Quiescent human aortic SMC in culture were exposed to LDL (40 microg protein/ml) alone or with washed human platelets (5 x 10(6)/ml), thrombin (40 units/ml), or a combination of these agents for 48 h. The media were removed, and both media and cell lysate fractions were assayed for malondialdehyde (MDA) content as an index of oxidation. Isolated platelets exposed to LDL and thrombin were studied in a similar manner to determine their individual oxidative activity. Finally, SMC and platelets were incubated with LDL and varying concentrations of thrombin (10-80 units/ml), both alone and in the presence of the thrombin inhibitors hirudin (u/u), and heparin (u/u), and MDA was measured. RESULTS: SMC and platelets each demonstrated an ability to oxidize LDL, increasing MDA concentrations by 1.8- (P < 0.05) and 4- (P < 0.01) fold, respectively, compared to lipid-free media. Both platelets (P < 0.05) and thrombin (P < 0.001) enhanced the oxidation of LDL by SMC, while a combination of these two agents resulted in an additive effect (P < 0.001). The SMC lysate fraction showed an increase in oxidative products following exposure to platelets (P < 0.01) but not thrombin, suggesting that platelets stimulated uptake of the oxidized lipid by the SMC. Isolated platelets responded to thrombin with an increase in MDA within the media (P < 0.001). Smooth muscle cells exposed to thrombin also showed a dose-dependent increase in LDL oxidation (P < 0.01). This effect was not altered by hirudin, but was significantly inhibited by heparin (P < 0.05). CONCLUSIONS: These results indicate that the oxidative potential of SMC and platelets is enhanced by their coincubation and by their concurrent exposure to thrombin. Heparin appears to block thrombin-stimulated oxidation. This interaction could be relevant to the dynamic interaction between atherosclerosis and thrombogenesis. PMID- 11855917 TI - Changes in the microvascular architecture of colorectal liver metastases following the administration of SMANCS/lipiodol. AB - BACKGROUND: Liver metastases are the major cause of death for patients with colorectal cancer. Surgical resection is at present the only curative option. Styrene maleic acid neocarzinostatin [SMANCS/Lipiodol (S/L)] targets the unique vascular architecture of tumor blood vessels, which are hyperpermeable and lack a well-developed lymphatic system. Here we report changes in the microvascular architecture of liver metastases by scanning electron microscopy (SEM) following the administration of S/L. MATERIALS AND METHODS: Liver metastases were induced by the intrasplenic injection of dimethylhydrazine induced colon cancer cells in mice. In this model tumor angiogenesis occurs at day 10, while exponential tumor growth occurs at day 16. Changes in the tumor microvasculature were observed at 3 weeks following treatment with S/L at these time points by SEM of corrosion casts. RESULTS: Tumors treated with S/L at day 10 appear similar to day 10 controls. Tumor vessels, 50 +/- 18 microm in diameter, are easily identified from hepatic vessels. Within the hepatic sinusoids are avascular spaces, 144 +/- 60 microm in diameter, which correspond to tumor cell aggregates at the initial stages of growth. Similarly, day 16 treated tumors appear comparable to day 16 controls. These vessels are narrower (84 +/- 32 microm vs. 150 +/- 70 microm) than their control counterparts. This is in contrast to vessels (216 +/- 36 microm in diameter) of a complex nature at 3 weeks. CONCLUSIONS: S/L exerts a marked and immediate effect on the tumor microvessels at both the angiogenic and the exponential phases of tumor growth. This agent is effective at the microvascular level during inhibition of metastatic growth. PMID- 11855918 TI - Local pharmaceutical release from a new hydrogel implant. AB - BACKGROUND: Solid hydrogel polymers can act as reservoirs for controlled drug release. The object of this study was to quantify release kinetics for a single example of a class of uniquely structured hydrogels. The polymer of this study belongs to a class of permanent implants that release pharmaceuticals by diffusion from an entangled cross-linked matrix rather than by absorption of the implant by the body. The cross-linked matrix (CLM) of this study is biocompatible and polymerized in situ, forming a solid that is mechanically bonded to the implant site. It can seal tissue as well as deliver drugs at predetermined rates. We evaluated dye and antibiotic egress and assessed release kinetics and retention of antibiotic activity following elution from the CLM. MATERIALS AND METHODS: The prepolymerized test CLM was prepared in two ratios, 6 and 20% prepolymer, in an activating aqueous solution. Aqueous solutions were prepared from dyes of varying molecular weights. Aliquots of prepolymerized test CLM were allowed to solidify and dye release into the supernatant was quantified by spectrophotometry over a 168-h period. Antibiotic solutions were also employed to form solid CLMs. Tetracycline release over time was characterized by spectrophotometry. Antibiotic dosed solid CLMs were placed on agar plates streaked with Escherichia coli and incubated. Growth inhibition was assessed for each antibiotic. RESULTS: In the test CLM, dye and antibiotic release were found to be inversely related to molecular weight and consistent with a diffusion model. CLMs formed from aqueous solutions containing higher molecular weight dyes and antibiotics released those constituents more slowly than lower molecular weight constituents. This finding, as well as the effect on release rate under varying prepolymer concentration, was consistent with a diffusion release mechanism. Antibiotic released from the tissue sealant was shown to be potent by consistent inhibition of E. coli. CONCLUSIONS: Pharmaceutical release by a representative CLM was found to be controllable by varying the concentration of the pharmaceutical in the activating aqueous solution. The polymerization and release mechanisms did not degrade antibiotic biologic activity. CLMs may be a general class of biocompatible polymers that can locally deliver clinically useful biologics, the release kinetics of which are unaffected by the variability of implant absorption/inflammation mechanisms. PMID- 11855919 TI - Role of macrophage inflammatory protein 2 in acute lung injury in murine peritonitis. AB - BACKGROUND: Acute lung injury is a frequent extraabdominal complication of bacterial peritonitis, and neutrophil plays an important role in this lung damage. Macrophage inflammatory protein 2 (MIP-2) serves the same chemotactic function as IL-8 which is a potent neutrophil chemotactic factor in humans, and we investigated the role of MIP-2 associated with neutrophil recruitment in the lung of murine peritonitis. METHODS: Cecal ligation and puncture (CLP) were performed on mice. MIP-2 levels in blood and lung tissue, MIP-2 mRNA expression in lung tissue and bronchoalveolar lavage fluid (BALF), and CD11b expression on peripheral blood neutrophil and BALF cells were determined after CLP. In addition, we investigated the effect of anti-MIP-2 antibody on the lung injury associated with peritonitis. RESULTS: MIP-2 mRNA expression was observed in lung tissue after CLP and numerous neutrophils were accumulated in the lung under those conditions. Anti-MIP-2 antibody contributed to the inhibition of the CD11b expression and chemotaxis of pulmonary neutrophils, lung edema, and thus the reduction in peritonitis-related mortality. CONCLUSIONS: MIP-2 plays a pivotal role in neutrophil recruitment in the lung following peritonitis, and control of neutrophil accumulation in the lung by neutralizing MIP-2 is recommended as a new therapeutic approach to the lung damage associated with peritonitis. PMID- 11855920 TI - Prediction of distant metastases after curative surgery for rectal cancer. AB - BACKGROUND: This study was performed to define selection criteria for adjuvant therapy in rectal cancer. MATERIALS AND METHODS: An immunohistochemical analysis using nine monoclonal antibodies against CEA, CD15s, CD44v6, DCC, E-cadherin, EGF R, NM23, PAI-1, and P53 was performed on paraffin sections of two matched (age, gender, UICC stage [I-III], year of operation [1982-1991]) groups of patients (n = 2 x 64) with rectal carcinoma curatively treated by surgery alone. The two groups differed only with regard to metachronous distant metastatic spread. In order to exclude the influence of surgery, all patients had to meet the selection criterion "free of locoregional disease." Follow-up was prospective (median 80 months). Conventional staining procedures and immunohistochemical evaluation were used. Tumor grading and lymphatic and extramural venous invasion were also investigated. Analysis was performed with Fisher's exact test and Kaplan-Meier estimates of disease-free survival (log rank). The Cox model was used for multivariate analysis. RESULTS: In univariate analysis only grading (P < 0.001) and extramural venous invasion (P < 0.001) correlated significantly with metachronous metastases. In multivariate analysis, beside grading (P = 0.010) and extramural venous invasion (P = 0.011), CD15s (P = 0.042) was also of significance. All other immunohistochemical markers failed. CONCLUSIONS: The histopathological parameters grading and extramural venous invasion appear to be acceptable predictors of metachronous distant spread in curatively resected rectal cancer. In contrast to the immunohistochemical markers, grading seems to better reflect the individual tumor phenotype and its behavior. PMID- 11855921 TI - Porcine hepatic phospholipid efflux during reperfusion after cold ischemia. AB - BACKGROUND: Cold preservation produces hepatic injury that is difficult to assess during early reperfusion. The value of reperfusion plasma choline phospholipid in predicting subsequent organ function is documented in these studies. MATERIALS AND METHODS: Livers of female Yorkshire pigs were prepared for transplantation. After 2 h of cold ischemia the reperfusion plasma was evaluated for choline phospholipid and cholesterol. These values were correlated with bile secretion, hepatic hemodynamics, oxygen uptake, and plasma sorbitol dehydrogenase levels. RESULTS: The isolated porcine liver demonstrates a rapid efflux of choline phospholipids into plasma during early reperfusion after cold preservation. After this initial efflux no subsequent plasma increment occurred. These choline phospholipid increments were isolated in plasma higher density (d > 1.063) lipoproteins and were not accompanied by equivalent increases in cholesterol. Neither biliary reflux nor lecithin cholesterol acyl transferase abnormalities contributed appreciably to the phospholipid increments in reperfusion plasma. Livers with the largest efflux of choline phospholipids had the most impaired circulatory and bile secretory function at 4 h of reperfusion. CONCLUSION: The immediate increase of choline phospholipids, particularly lysophosphatidylcholine, in reperfusion plasma after cold ischemia provides an index of the injury occurring during this interval and correlates with early organ function. PMID- 11855922 TI - Surgical site infections: reanalysis of risk factors. AB - BACKGROUND: Surgical site infections (SSI) are the most common nosocomial infection in surgical patients, accounting for 38% of all such infections, and are a significant source of postoperative morbidity resulting in increased hospital length of stay and increased cost. During 1986-1996 the Center for Disease Control and Prevention's National Nosocomial Infections Surveillance system reported 15,523 SSI following 593,344 operations (2.6%). Previous studies have documented patient characteristics associated with an increased risk of SSI, including diabetes, tobacco or steroid use, obesity, malnutrition, and perioperative blood transfusion. In this study we sought to reevaluate risk factors for SSI in a large cohort of noncardiac surgical patients. METHODS: Prospective data (NSQIP) were collected on 5031 noncardiac surgical patients at the Veteran's Administration Maryland Healthcare System from 1995 to 2000. All preoperative risk factors were evaluated as independent predictors of surgical site infection. RESULTS: The mean age of the study cohort was 61 plus minus 13. SSI occurred in 162 patients, comprising 3.2% of the study cohort. Gram-positive organisms were the most common bacterial etiology. Multiple logistic regression analysis documented that diabetes (insulin- and non-insulin-dependent), low postoperative hematocrit, weight loss (within 6 months), and ascites were significantly associated with increased SSI. Tobacco use, steroid use, and chronic obstructive pulmonary disease (COPD) were not predictors for SSI. CONCLUSION: This study confirms that diabetes and malnutrition (defined as significant weight loss 6 months prior to surgery) are significant preoperative risk factors for SSI. Postoperative anemia is a significant risk factor for SSI. In contrast to prior analyses, this study has documented that tobacco use, steroid use, and COPD are not independent predictors of SSI. Future SSI studies should target early preoperative intervention and optimization of patients with diabetes and malnutrition. PMID- 11855923 TI - Captopril inhibits secretin-induced pancreatic bicarbonate output. AB - BACKGROUND: Renin-angiotensin systems function at both the organ and systemic levels. Previous studies suggest that angiotensin II (Ang II) stimulates pancreatic secretion in vitro. In contrast, in vivo studies suggest that Ang II inhibits pancreatic secretion. To further assess Ang II's influence on pancreatic secretion, we examined the effect of captopril on secretin-stimulated pancreatic output. METHODS: After a 30-min basal period, four conscious dogs with chronic gastric and Herrera pancreatic fistulas received an intravenous bolus of captopril (0.1 mg/kg) followed by a continuous infusion (25 microg/kg/min). Control studies were performed with volume- and rate-matched saline infusion. After 1 h, secretin infusion was begun at 16 ng/kg/h, doubling the dose every 30 min. Pancreatic juice was analyzed for bicarbonate and protein. A paired t test was used to assess statistical significance. RESULTS: When compared to controls, pancreatic bicarbonate outputs were lower during captopril administration; the difference between captopril and control was statistically significant at the highest secretin dose. Protein outputs also appeared lower during captopril administration, although these differences were not statistically significant. CONCLUSION: These data suggest that Ang II may augment secretin-induced pancreatic secretion. Further, the data seemingly refute the inhibitory role attributed to Ang II in earlier studies. PMID- 11855924 TI - Multiple heparanases are expressed in polymorphonuclear cells. AB - BACKGROUND: Heparan sulfate proteoglycans are complex cell surface molecules containing polysaccharides called heparan sulfate. Lysosomes, platelet granules, and neutrophils (polymorphonuclear cells) contain heparanases that degrade heparan sulfate. There are at least two groups of heparanases: connective tissue activating-peptide (CTAP-III) and mammalian heparanase (hpa). The purpose of this study was to quantify the expression of both CTAP-III and hpa in neutrophils and their heparanase activity. MATERIALS AND METHODS: Neutrophils were isolated from whole blood, total RNA collected, and reverse transcriptase--polymerase chain reaction (RT-PCR) performed. Primers were designed for CTAP-III and hpa-1 sequences from GenBank. Neutrophil lysate underwent Western blot analysis (and quantification) with antibodies to the C-terminus of CTAP-III and the 50-kDa subunit of hpa1. Chromatography separated these components of lysate, which were then tested for heparanase activity. RESULTS: Both CTAP-III (281 bp) and hpa-1 (485 bp) messenger RNA (mRNA) were expressed equally by neutrophils with use of quantitative RT-PCR. By Western blot analysis, a CTAP-III-like protein was detected at 80 kDa, and hpa-1 was detected as a 50-kDa protein, with expression not significantly different (P > 0.05). Heparanase activity was significantly different (P < 0.0001) for the 50-kDa hpa-1 protein (1.51 x 10(-6) micromol/min) and the 80-kDa CTAP-III-like protein (0.85 x 10(-6) micromol/min). CONCLUSIONS: Human neutrophils express mRNA and protein for both a CTAP-III-like protein and hpa-1. Although expressed in similar quantity for mRNA and protein, Hpa-1 was more active as heparanase than the CTAP-III-like protein. With more than one class of heparanase in their granules, neutrophils may be able to modify different kinds of heparan sulfate chains. PMID- 11855925 TI - ICG pulse spectrophotometry for perioperative liver function in hepatectomy. AB - BACKGROUND: The indocyanine green (ICG) clearance test has been used to estimate liver functional reserve before hepatectomy. However, changes in ICG clearance after hepatectomy have not been investigated, and their extent remains unknown. PATIENTS AND METHODS: The ICG(K) value, signifying the ICG elimination rate constant, was measured with pulse-dye densitometry before operation and 1, 2, 3, 5, and 7 days postoperatively in 22 patients who underwent liver resection of various extent. CT volumetry was used to calculate the residual liver volume ratio. The relationship between the pre- and postoperative ICG(K) value and the residual liver volume ratio was examined statistically. RESULTS: There was a significant drop in ICG(K) value, from 0.193 +/- 0.011 before operation to 0.160 +/- 0.013 on Postoperative Day 1, and then it remained significantly low at the postoperative examination times. The residual liver volume ratio was 70.2 +/- 5.4%. The estimated ICG(K) value, calculated by the preoperative ICG(K) value and the residual liver volume ratio, showed a significant correlation with the actual postoperative value (r = 0.859 on Postoperative Day 1, P < 0.0001). In five patients with prolonged jaundice, the estimated ICG(K) value was significantly lower than in those without it (0.077 +/- 0.028 versus 0.153 +/- 0.012, P = 0.0136). CONCLUSIONS: The perioperative ICG(K) value measured by pulse-dye densitometry revealed a significant decrease in ICG(K) after operation depending on the reduction in liver volume, and the estimated ICG(K) based on the residual liver volume was useful in predicting postoperative morbidity. PMID- 11855926 TI - Spatially selective laser coagulation of the gastric wall: a new methodology. AB - BACKGROUND: The incidence of lymph node metastasis is low in early gastric cancer. Early cancer without nodal metastasis may be theoretically eradicated by local treatment alone. However, except for gastrectomy, there exists no reliable treatment for submucosal cancer. We present a new laser system which irradiates the gastric wall from the serosal side combined with synchronous serosal cooling that can produce deep thermal coagulation without transmural damage. METHODS: A laser handpiece with a built-in surface cooling system has been devised, employing a diode laser of 980 nm wavelength. Animal experiments were conducted to ascertain whether this laser system could coagulate the mucosa and submucosa with acceptable injury to the muscular layer and serosa. The gastric wall was irradiated from the serosal side with synchronous serosal cooling at 6.0-10.0 W power output and 50-400 s exposure time. The degree and depth of damage were histologically assessed after 7 days. RESULTS: Ninety-one points were irradiated in 8 dogs. Mucosal coagulation was observed during the treatment while the serosa showed no serious injury. No gastric perforation occurred up to 7 days later. Histologically, the damage did not penetrate the gastric wall except for cases where 10.0 W and 400 s were used. Cauterization of the mucosa and submucosa with acceptable muscle layer damage was achieved in selected settings. CONCLUSIONS: This laser system enables thermal coagulation of the gastric mucosa and submucosa with acceptable muscle layer damage. This makes it a promising, novel, minimally invasive treatment for submucosal cancer. PMID- 11855927 TI - Human interleukin 10 gene therapy decreases the severity and mortality of lethal pancreatitis in rats. AB - BACKGROUND: Studies have proven the validity of interleukin-10 (IL-10) in the treatment of experimental pancreatitis. Prophylactic human IL-10 (hIL-10) gene treatment attenuated the severity in cerulein models. Our research aims to study whether the therapeutic hIL-10 gene could decrease both severity and mortality in a lethal pancreatic model. METHODS: Severe acute pancreatitis (SAP) was induced by sodium taurocholate. A plasmid-hIL-10 construct (pcDNA3-hIL-10) complexed with cationic liposomes was administered to SAP rats by a single intraperitoneal injection. Levels of hIL-10 in the pancreas, liver, and lungs were determined by ELISA kits. The severity of pancreatitis was assessed in terms of serum amylase, histology, and tissue tumor necrosis factor alpha (TNF-alpha). Mortality, observed for 7 days, was evaluated for gene therapy or control groups. RESULTS: After hIL-10 gene therapy, hIL-10 levels in the pancreas, liver, and lungs increased significantly and the serum amylase, tissue TNF-alpha, and histological changes in pancreas, liver, and lungs decreased markedly. Therefore, mortality was significantly reduced in the hIL-10 gene therapy group, in which 70% of rats survived in the 7-day observation, while only 10% survived in untreated groups (P < 0.05). CONCLUSION: We found that liposome/hIL-10 gene therapy decreased severity and mortality in SAP, even carried out after SAP establishment, predicting a more convenient shift to clinical applications. PMID- 11855928 TI - OPA1 (Kjer type) dominant optic atrophy: a novel mitochondrial disease. AB - Dominant optic atrophy (DOA) is the most common form of inherited optic neuropathy. Although heterogeneous, a major locus has been mapped to chromosome 3q28 and the responsible gene, OPA1, was recently identified. OPA1 is a mitochondrial dynamin-related GTPase implicated in the formation and maintenance of the mitochondrial network. To date, 62 mutations have been identified in a total of 201 DOA patients. Most of them (90%) are distributed from exons 8 to 28 with a majority in the GTPase domain (54%). None were found in the alternatively spliced exons 4, 4b, and 5b. Half of them are truncative mutations (50%) with a frequent recurrent allele, c.2708delTTAG. Most missense mutations (81%) cluster within the putative GTPase domain. Various pathogenic mechanisms may play a role in OPA1 DOA. Truncative mutations in the N-terminal region and perhaps missense mutations in the GTPase domain lead to a loss of function of the encoded protein and haplotype insufficiency. However, there is a cluster of truncation mutations in the in C-terminus, a putative dimerization domain, that could act through a dominant negative effect. The findings that OPA1-type DOA, as Leber optic neuropathy, is caused by the impairment of a mitochondrial protein address the question of the vulnerability of the retinal ganglion cell in response to mitochondrial defects. PMID- 11855929 TI - Straight-chain acyl-CoA oxidase knockout mouse accumulates extremely long chain fatty acids from alpha-linolenic acid: evidence for runaway carousel-type enzyme kinetics in peroxisomal beta-oxidation diseases. AB - Extremely long chain polyunsaturated fatty acids (ELCPs) with >24 carbons and four or more double bonds are normally found in excitatory tissues but have no known function, and are greatly increased in brain and other tissues of humans with peroxisomal disorders. Straight-chain acyl-CoA oxidase (AOX) catalyzes the first, rate-limiting step of peroxisomal beta-oxidation of very-long-chain saturated and unsaturated fatty acids. We have studied the polyunsaturated fatty acid metabolism of AOX knockout mice (AOX-/- as a model of human AOX deficiency (pseudo-neonatal adrenoleukodystrophy), and as a genetic tool to test the putative peroxisomal beta-oxidation involvement in polyunsaturated fatty acid synthesis. Liver lipids of 26-day-old weanling AOX-/- mice livers accumulate n-3 and n-6 ELCPs from C24 to C30 with 5 and 6 double bonds, have 356 +/- 66 microg/g docosahexaenoic acid (22:6n-3), similar to congenic (AOX -/* = AOX+/+ and AOX+/-) controls (401 +/- 96 microg/g), but increased 22:5n-6 (22.4 +/- 3.7 vs 6.4 +/- 1.5 microg/g). AOX+/* mice injected intraperitoneally at 23 days with [U-(13)C] 18:3n-3 show strong labeling of 22:6n-3 after 72 h, whereas AOX -/- mice display less labeling of 22:6n-3 but strong tracer incorporation into 24:6n-3, 26:6n-3, and 28:6n-3, after the same period. These data suggest that ELCPs are natural runaway elongation by-products of 22:6n-3 and 22:5n-6 synthesis, which are normally disposed of by peroxisomal beta-oxidation. Under conditions with impaired peroxisomal beta-oxidation, such as Zellweger syndrome and adrenoleukodystrophies, ELCPs accumulate due to increased synthesis and impaired disposal. Two mechanisms for the formation of these runaway elongation by products and the involvement of secondary carnitine deficiency in this process are proposed: n-3 ELCPs are synthesized by a carnitine-dependent multifunctional mitochondrial docosahexaenoic acid synthase (mtDHAS) which normally synthesizes primarily 22:6n-3, while n-6 ELCPs are synthesized by independent elongation enzymes in the endoplasmic reticulum. PMID- 11855930 TI - Early neonatal diagnosis of long-chain 3-hydroxyacyl coenzyme a dehydrogenase and mitochondrial trifunctional protein deficiencies. AB - Tandem mass spectrometry (MS/MS) has been introduced in several newborn screening programs for the detection of a large number of inborn errors of metabolism, including fatty acid oxidation disorders (FAOD). Early identification and treatment of FAOD have the potential to improve outcome and may be life-saving in some cases; an estimated 5% of sudden infant deaths are attributable to undiagnosed disorders of fatty acid oxidation. We report very early neonatal presentations of long-chain 3-hydroxyacyl-CoA dehydrogenase (LCHAD) and mitochondrial trifunctional protein (TFP) deficiencies confirmed by molecular analysis. Both patients had cardiorespiratory collapse and hypoglycemia, without a history of maternal pregnancy complications. Retrospective MS/MS analysis of the original newborn screening blood spots revealed characteristic acylcarnitine profiles. These cases are among the earliest reported presentations of LCHAD and TFP deficiencies and further illustrate the potential of MS/MS as a valuable tool for newborn screening of FAOD. However, timely analysis and reporting of results to clinicians are essential, because these disorders can manifest in the first few days of life. PMID- 11855931 TI - Expression of CTNS alleles: subcellular localization and aminoglycoside correction in vitro. AB - Mutations in CTNS result in one of three forms of cystinosis: benign, intermediate, or nephropathic. Homozygosity for a nonsense mutation in CTNS (753G -->A), encoding a premature termination codon (PTC) at amino acid 138 (W138X), results in nephropathic cystinosis. Gentamicin is known to induce PTC readthrough and hence full-length protein production. We demonstrate that addition of gentamicin (300 microg/ml) to cystinotic fibroblasts leads to depletion of intracellular cystine in cell lines with a premature termination codon, but not in those with a large deletion or a deletion leading to a frameshift mutation. Plasmids were constructed with GFP as a C-terminal or N-terminal fusion to CTNS. The normal CTNS protein fused with either N- or C-terminal GFP colocalized with Lysotracker red, a fluorescent stain which selectively accumulates in lysosomes. PTC-GFP, a construct with GFP fused to the C-terminus of CTNS containing a PTC, allowed GFP to serve as a reporter of PTC readthrough. No significant fluorescence was observed in PTC-GFP-transfected cells in the absence of gentamicin but was seen and localized to lysosomes in its presence. A patient with a splice site mutation (IVS11 + 2T -->C) that eliminates the GYDQL lysosomal targeting sequence of cystinosin on one allele, and a PTC mutation (753G -->A) on the other, displays the intermediate phenotype. Transfection of the splice site mutant allele into CTNS null fibroblasts produced cystine depletion. Plasmids with GFP fused to the N-terminus of CTNS containing the splice site mutation (GFP SS) were constructed. While the normal CTNS-GFP fusion protein was found to colocalize with Lysotracker red almost exclusively, the GFP-SS fusion product was found in the plasma membrane and cytoplasm, as well as lysosomes. A second lysosomal targeting motif in CTNS is present in this sequence, just proximal to the mutation, accounting for the partial lysosomal localization. PMID- 11855932 TI - A novel intronic mutation results in the use of a cryptic splice acceptor site within the coding region of UGT1A1, causing Crigler-Najjar syndrome type 1. AB - Crigler-Najjar syndrome type 1 (CN-1) is characterized by severe unconjugated hyperbilirubinemia due to an inherited deficiency of hepatic bilirubin uridinediphosphoglucuronate glucuronosyltransferase (UGT1A1), inherited as an autosomal recessive characteristic. CN-1 is potentially lethal because of the risk of bilirubin encephalopathy (kernicterus). Genetic lesions of the coding region of the UGT1A1 gene are known to cause CN-1. Here, we report a CN-1 patient who has a novel G > A mutation at the splice acceptor site in intron 4 (IVS4-1 G > A) on one allele, and a T > A substitution followed by a 13-nt deletion in exon 2 (877T > A 878-890del) of the other allele. As the UGT1A1 gene is expressed specifically in the liver, structural analysis of the expressed UGT1A1 mRNA requires liver biopsy. To use a noninvasive approach to determine the effect of the splice site mutation on splicing of the RNA transcript, we amplified the relevant region of the genomic DNA by long-range polymerase chain reaction (PCR). The amplicon was cloned in an expression plasmid and transfected into COS-7 cells. The expressed mRNA was amplified by reverse-transcription-primed PCR. Nucleotide sequence determination of the amplicon showed that the splice acceptor site mutation caused splicing of the 3'-end of exon 4 to a cryptic splice site within exon 5. This resulted in deletion of the first 7 nucleotides of exon 5, causing a frameshift and premature truncation of UGT1A1, with consequent inactivation of the enzyme. PMID- 11855933 TI - Denaturing gradient gel electrophoresis analysis of the tissue nonspecific alkaline phosphatase isoenzyme gene in hypophosphatasia. AB - Hypophosphatasia, a heritable form of rickets/osteomalacia, was first described in 1948. The biochemical hallmark, subnormal alkaline phosphatase (ALP) activity in serum, reflects a generalized disturbance involving the tissue-nonspecific isoenzyme of ALP (TNSALP). Deactivating mutations in the gene that encodes TNSALP have been reported in patients worldwide. Nevertheless, hypophosphatasia manifests an extraordinary range of clinical severity spanning death in utero to merely premature loss of adult teeth. There is no known medical treatment. To delineate the molecular pathology which explains the disease variability and to clarify the pattern(s) of inheritance for mild cases of hypophosphatasia, we developed comprehensive mutational analysis of TNSALP. High efficiency of mutation detection was possible by denaturing gradient gel electrophoresis (DGGE). Primers and conditions were established for all TNSALP coding exons (2 12) and adjacent splice sites so that the amplicons incorporated a GC clamp on one end. For each amplicon, the optimum percentage denaturant was determined by perpendicular DGGE. In 19 severely affected pediatric subjects (having perinatal or infantile hypophosphatasia or early presentation during childhood) from among our large patient population, we detected 2 TNSALP mutations each in 16 patients (84%) as expected for autosomal recessive disease. For 2 patients (11%), only 1 TNSALP mutation was detected by DGGE. However, one subject (who died from perinatal hypophosphatasia) had a large deletion as the second mutation. In the other (with infantile hypophosphatasia), no additional mutation was detected by DNA sequencing of all protein-coding exons. Possibly, she too has a deletion. For the final patient, with unclassifiable hypophosphatasia (5%), we detected only a single mutation which has been reported to cause relatively mild autosomal dominant disease; the other allele appeared to be intact. Hence, DGGE analysis was 100% efficient in detecting mutations in the coding exons and adjacent splice sites of TNSALP in this group of severely affected patients but, as expected, failed to detect a large deletion. To date, at least 78 different TNSALP mutations (in about 70 hypophosphatasia patients) have been reported globally. In our large subset of severely affected patients, we identified 8 novel TNSALP mutations (Ala34Ser, Val111Met, Delta G392, Thr117His, Arg206Gln, Gly322Arg, Leu397Met, and Gly409Asp) and 1 new TNSALP polymorphism (Arg135His) furthering the considerable genotypic variability of hypophosphatasia. PMID- 11855934 TI - Insulin reduces serum glycosylphosphatidylinositol phospholipase D levels in human type I diabetic patients and streptozotocin diabetic rats. AB - The enzyme glycosylphosphatidylinositol phospholipase D has a postulated role in the insulin-mimetic signaling pathway of glycosylphosphatidylinositol compounds. We have investigated enzyme activity in the serum of human type I diabetic patients and plasma and tissues of streptozotocin-induced diabetic rats following insulin administration. In the human diabetic patients serum enzyme activity fell by an average of 10.6% (SEM = 2.7; P = 0.008; n = 20) following administration of insulin. In addition serum enzyme activity appeared to be depleted by 27% (SEM = 8.8; P = 0.011; n = 10) compared to nondiabetic controls. In untreated diabetic rats plasma enzyme activity gradually increased 0.3-fold over a 6-week period (P < 0.001; n = 8), this increase was reversed and activity normalized when these animals were treated with insulin. Cloning of the rat glycosylphosphatidylinositol phospholipase D cDNA enabled confirmation of the liver as the principal organ of synthesis. Analysis of mRNA levels in the livers of the diabetic rats showed that gene expression was reduced in the insulin treated animals compared to the noninsulin-treated controls by 0.7-fold (P = 0.004; n = 4). Tissue enzyme activity was also reduced in the insulin-treated rats; in skeletal muscle enzyme activity was 0.3-fold lower (P = 0.001; n = 4). Insulin therefore decreases glycosylphosphatidylinositol phospholipase D synthesis in diabetic animals resulting in decreased serum enzyme levels, suggesting a relationship between this enzyme and the function of insulin. PMID- 11855935 TI - Metabolism of D-[1-(13)C]fructose, D-[2-(13)C]fructose, and D-[6-(13)C]fructose in rat hepatocytes incubated in the presence of H(2)O or D(2)O. AB - Isolated hepatocytes from fed rats were exposed for 120 min to D-[1 (13)C]fructose, D-[2-(13)C]fructose, or D-[6-(13)C]fructose in the presence of H(2)O or D(2)O. The identification and quantification of (13)C-enriched metabolites (D-glucose, L-lactate) in the incubation medium and the measurement of their deuterated isotopomers indicated that the ketohexose was phosphorylated predominantly at the intervention of fructokinase and that the majority of the D glyceraldehyde molecules generated from d-fructose 1-phosphate were further metabolized, e.g., after phosphorylation to D-glyceraldehyde 3-phosphate. It is proposed that the present procedure may help to further characterize the regulation of D-fructose metabolism in both hepatocytes and other cell types. PMID- 11855936 TI - Site-directed mutations in the FAD-binding domain of glycerophosphate dehydrogenase: catalytic defects with preserved mitochondrial anchoring of the enzyme in transfected COS-7 cells. AB - Single-strand conformational polymorphism analysis of mitochondrial FAD-linked glycerophosphate dehydrogenase (mGDH) gene has revealed mutations in both the calcium- and FAD-binding domains of this enzyme in some diabetic patients. It was now investigated whether site-directed mutations in the FAD-binding domain of the mGDH gene may affect the mitochondrial anchoring and catalytic activity of the enzyme. COS-7 cells were transfected with plasmid cDNA coding for either wild type or mutated human mGDH (G --> A substitutions at positions 352, 355, and 364 and A --> C substitution at position 390) fused, when required, at the N-terminus of green fluorescent protein. The activity of mGDH was measured by both radioisotopic ((3)HOH production from l-[2-(3)H]glycerol 3-phosphate) and colorimetric (iodoformazan formation) procedures. In cells transfected with the mGDHwt-EGFP or mGDHmut-EGFP constructs, the fused protein was found by confocal microscopy exclusively in the mitochondria, colocalized with a mitochondrial marker. In homogenates of COS-7 cells transfected with mGDHmut, however, the catalytic activity of the enzyme was decreased, this coinciding with low ratios between both the activities measured in the absence/presence of exogenous FAD and the results obtained by the colorimetric/radioisotopic procedure. Thus, although the present site-directed mutations of the mGDH gene failed to impair the mitochondrial anchoring of the enzyme, they led to catalytic defects that were, in some respect, comparable to those previously encountered in the lymphocytes or islets of type 2 diabetic patients. PMID- 11855937 TI - Detection of sepiapterin in CSF of patients with sepiapterin reductase deficiency. AB - Sepiapterin reductase (SR) deficiency was recently described in patients with a severe biogenic amine deficiency presenting without hyperphenylalaninemia and it was suggested that the tetrahydrobiopterin (BH(4)) pathway may be different in different cells and tissues. We now developed a HPLC method for the measurement of yellow fluorescing sepiapterin for the rapid diagnosis of SR deficiency. Sepiapterin was elevated in CSF from two patients with SR deficiency (5.6 and 11.4 nmol/L) when compared with healthy controls (<0.5 nmol/L). Our data further support the hypothesis that sepiapterin is an intermediate in the salvage pathway of BH(4) and that it accumulates in the brain of patients with SR deficiency. PMID- 11855938 TI - Novel donor splice site mutation of ABCG5 gene in sitosterolemia. AB - In a patient with sitosterolemia, we found two different mutations of the ATP binding cassette, subfamily G, member 5 (ABCG5) gene. The first is a missense mutation that changes the amino acid residue at position 419 from arginine to histidine, i.e., R419H. The second is a novel splicing mutation affecting the invariant guanine at the first base of the donor splice site of intron 12, i.e., IVS12 + 1G --> A. The father of the patient is heterozygous for the missense mutation, and the mother is heterozygous for the splicing mutation. No mutations were found in the sister of the patient. Up until now, the missense mutation has only been found in Japanese patients with sitosterolemia. We believe that R419H in our Chinese patient may have the same origin as the mutation in the Japanese patients with sitosterolemia. PMID- 11855939 TI - A novel nonsense mutation (515del4) in muscle carnitine palmitoyltransferase II deficiency. AB - We identified a novel nonsense mutation in the carnitine palmitoyltransferase (CPT; EC 2.3.1.21) II gene in a patient with biochemical evidence of CPT II deficiency. The 39-year-old man suffered from the muscle form of CPT II deficiency. Attacks of myalgia and muscle weakness started in childhood and led to renal failure five times. A mild proximal weakness of the lower limbs was left as a residue. Molecular genetic analysis revealed the common S113L mutation on one allele. On the other allele a novel 4-bp deletion starting at codon 515 (515del4) was found leading to frameshift that results in a stop codon 15 codons upstream. Our data further expand the genetic heterogeneity in patients with CPT II deficiency. PMID- 11855940 TI - Tetrahydrobiopterin-responsive phenylalanine hydroxylase deficiency: possible regulation of gene expression in a patient with the homozygous L48S mutation. PMID- 11855941 TI - Streptococcus sanguis secretes CD14-binding proteins that stimulate cytokine synthesis: a clue to the pathogenesis of infective (bacterial) endocarditis? AB - Streptococcus sanguis is the major causative organism of infective (bacterial) endocarditis but, surprisingly, almost nothing is known about how it induces endocardial inflammation. In earlier studies we have shown that many bacteria secrete potent cytokine-inducing or -inhibiting proteins. We have therefore isolated the material secreted by S. sanguis grown on blood agar or in broth culture and have tested its ability to induce human peripheral blood monocytes to synthesize pro-inflammatory cytokines. The activation of monocytes by the secreted components of S. sanguis was almost totally blocked by heat and trypsin treatment but not by the lipopolysaccharide-inactivating antibiotic, polymyxin B, suggesting that activity is due to secreted proteins. The activity of the secreted material was significantly reduced by anti-CD14 monoclonal antibodies suggesting that the active protein (or proteins) was binding to the CD14/Toll like receptor (TLR)4 complex. Fractionation of the secreted proteins by high performance liquid chromatography (HPLC) identified two proteins as being responsible for the majority of the cytokine induction: a manganese-dependent superoxide dismutase and a 190 kDa protein, which could not be sequenced, but which was neither CshA nor the PI/II proteins. These proteins, or the receptors to which they bind, may be therapeutic targets and may allow the development of adjunctive therapies to prevent endocardial damage during the often prolonged treatment of infective endocarditis with antibiotics. In addition, blocking of CD14 may have some therapeutic benefit. PMID- 11855942 TI - L12 enhances gonococcal transcytosis of polarized Hec1B cells via the lutropin receptor. AB - We previously reported that gonococci convert to a more invasive phenotype (Inv(+)GC) following contact with cells expressing the lutropin receptor (LHr) and that Inv(+)GC express a novel adhesin that interacts with LHr. We propose that this adhesion allows Inv(+)GC to activate LHr and induce gonococcal transcytosis, usurping normal LHr function in fallopian and endometrial epithelium, which is to transport fetal chorionic gonadotropin (hCG) into the mother. Infected polarized Hec1B monolayers, grown on collagen-coated transwells, showed that the passage of GC across the monolayer occurred rapidly, within 30 min, and proceeded at a constant rate with Inv(+)GC passage three-fold faster than GC grown in tissue culture media alone (Inv(-)GC). Electron microscopy found that Inv(+)GC triggered pseudopod formation around the bacterium, with GC found throughout the Hec1B targets within 30 min, while Inv(-)GC did neither. Pre treatment of Inv(-)GC with recombinant ribosomal protein L12, a gonococcal "hCG like" protein previously shown to increase invasion, also increased Inv(-)GC transcytosis to the rate of Inv(+)GC. This enhancement was completely abolished by addition of luteinizing hormone, a cognate ligand of LHr. This is convincing evidence that surface expressed L12 mediates gonococcal invasion and transcytosis via LHr, a mechanism that could be important in the development of invasive gonococcal disease in women. PMID- 11855943 TI - Different host defences are required to protect mice from primary systemic vs pulmonary infection with the facultative intracellular bacterial pathogen, Francisella tularensis LVS. AB - Francisella tularensis is a zoonotic, facultative intracellular bacterial pathogen capable of initiating infection, tularemia, via multiple routes including dermal micro-abrasions and inhalation. Mouse models of systemically initiated infection with F. tularensis LVS have been used extensively to reveal potential host defence mechanisms against the pathogen. Such studies have demonstrated the critical need for neutrophils and interferon-gamma (IFN-gamma) to combat the early stages of primary experimental tularaemia initiated by systemic routes. Surprisingly, however, the present study shows that these defences appear not to combat early pulmonary tularaemia initiated by inhalation of the pathogen into the lower airways. The results imply that the effectiveness of particular anti-bacterial host defences vary with invasion site. Thus, it is impossible to predict effective host defence mechanisms against inhalation initiated tularaemia from current knowledge of anti- Francisella defences that have been shown to combat systemically-initiated infection. PMID- 11855944 TI - Role of immune sera in the in-vitro phagocytosis of Bordetella pertussis strains. AB - In this study, phagocytosis of Bordetella pertussis was assessed using a human monocyte-derived macrophage line (THP-1) and immune sera from children who had received primary vaccination during the Italian clinical trial on the efficacy of two acellular three-component (PT-FHA-PRN) and one whole-cell pertussis vaccines. The results demonstrate that phagocytosis of opsonized bacteria with specific immune sera is not significantly enhanced compared with that of non-opsonized bacteria or bacteria opsonized with non-immune sera. A similar result was obtained also using B. pertussis strains showing variants of the pertactin antigen suggesting that those variations do not reduce the capability of the bacterium to invade the monocytes. PMID- 11855945 TI - Detection of Pseudomonas aeruginosa cell-to-cell signals in lung tissue of cystic fibrosis patients. AB - Chronic Pseudomonas aeruginosa infections lead to progressive lung tissue destruction in cystic fibrosis (CF) patients. Two bacterial cell-to-cell signals, 3-oxo-C(12)-HSL and C(4)-HSL are required for the production of several extracellular virulence factors. 3-oxo-C(12)-HSL is also required for the development of a differentiated biofilm, induces IL-8 production by epithelial cells and possesses immunomodulatory activities. These two signalling molecules are therefore believed to play a role in the pathogenesis of P. aeruginosa infections, but have never been isolated from infected human tissues. We extracted and quantified the two P. aeruginosa cell-to-cell signals from lung tissues of two CF patients infected by P. aeruginosa. 3-oxo-C(12)-HSL and C(4) HSL were detected in the lung tissues in fmol/gram, respectively pmol/gram concentrations; the ratio C(4)-HSL/3-oxo-C(12)-HSL exceeded 100 in all tissue samples. Random Amplified Polymorphism DNA genotyping revealed that one genotype was present per lung. In vitro the P. aeruginosa isolates from the two lungs produced 3-oxo-C(12)-HSL, whereas some isolates did not produce detectable C(4) HSL. Our results suggest that both P. aeruginosa cell-to-cell signals were produced in the lung tissue of these two cystic fibrosis patients. PMID- 11855947 TI - Locating genes by linkage and association. PMID- 11855948 TI - Reconstructing parental genotypes when testing for linkage in the presence of association. AB - Various family-based association methods have recently been proposed that allow testing for linkage in the presence of linkage disequilibrium between a marker and a disease even if there is only incomplete parental-genotype information. For some families, it may be possible to reconstruct missing parental genotypes from the genotypes of their offspring. Treating such a reconstructed family as if parental genotypes have been typed, however, can introduce bias. The reconstruction-combined transmission/disequilibrium test (RC-TDT) and its X chromosomal counterpart, XRC-TDT, employ parental-genotype reconstruction and correct for the biases involved in this reconstruction without relying on population marker allele frequencies. For the two tests, exact P values can be obtained by numerically calculating the convolution of the null distributions corresponding to the families in the sample. PMID- 11855949 TI - A train of thoughts on gene mapping. AB - Complex traits, by definition, are the pheonotypic outcome from multiple interacting genes. The traditional analysis of association studies on complex traits is to test one locus at a time, but a better approach is to analyze all markers simultaneously. We previously proposed a two-stage approach, first selecting the influential markers and then modeling main and interaction effects of these markers. Here we introduce alternative approaches to marker selection and discuss issues regarding analytical tools for disease gene mapping, marker selection, and statistical modeling. PMID- 11855950 TI - Genomic control, a new approach to genetic-based association studies. AB - During the past decade, mutations affecting liability to human disease have been discovered at a phenomenal rate, and that rate is increasing. For the most part, however, those diseases have a relatively simple genetic basis. For diseases with a complex genetic and environmental basis, new approaches are needed to pave the way for more rapid discovery of genes affecting liability. One such approach exploits large, population-based samples and large-scale genotyping to evaluate disease/gene associations. A substantial drawback to such samples is the fact that population heterogeneity can induce spurious associations between genes and disease. We describe a method called genomic control (GC), which obviates many of the concerns about population substructure by using the features of the genomes present in the sample to correct for stratification. Two such approaches are now available. The GC approach exploits the fact that population substructure generate "overdispersion" of statistics used to assess association. By testing multiple polymorphisms throughout the genome, only some of which are pertinent to the disease of interest, the degree of overdispersion generated by population substructure can be estimated and taken into account. The other approach, called Structured Association (SA), assumes that the sampled population, while heterogeneous, is composed of subpopulations that are themselves homogeneous. By using multiple polymorphisms throughout the genome, SA probabilistically assigns sampled individuals to these latent subpopulations. We review in detail the overdispersion GC. In addition to outlining the published ideas on this method, we describe several extensions: quantitative trait studies and case-control studies with haplotypes and multiallelic markers. For each study design our goal is to achieve control similar to that obtained for a family-based study, but with the convenience found in a population-based design. PMID- 11855951 TI - Methods for linkage analysis of quantitative trait loci in humans. AB - This paper reviews linkage analysis methods for detecting loci associated with quantitative traits in humans. All such methods are based on the underlying principle that family members who have similar trait values should have higher than expected levels of sharing of genetic material (identity by descent) near the genes that influence those traits. A number of different statistical methods for testing that association between shared trait values and shared identity by descent have been developed over the past 30 or more years. These different types of tests are reviewed here, with emphasis on their theory and derivations. Robustness and power are also discussed. PMID- 11855952 TI - Prospects for association-based fine mapping of a susceptibility gene for a complex disease. AB - The potential of association studies for fine-mapping loci with common disease susceptibility alleles for complex genetic diseases in outbred populations is unclear. For a battery of tightly linked anonymous genetic markers spanning a candidate region centered around a disease locus, simulation methods based on a coalescent process with mutation, recombination, and genetic drift were used to study the spatial distribution of markers with large noncentrality parameters in a case-control study design. Simulations with a disease allele at intermediate frequency, presumably representing an old mutation, tend to exhibit the largest noncentrality parameter values at markers near the disease locus. In contrast, simulations with a disease allele at low frequency, presumably representing a young mutation, often exhibit the largest noncentrality parameter values at markers scattered over the candidate region. In the former case, sample sizes or marker densities sufficient to detect association are likely to lead to useful localization, whereas, in the latter case, localization of the disease locus within the candidate region is much less likely, regardless of the sample size or density of the map. The simulations suggest that for a single marker analysis, the simple strategy of choosing the marker with smallest associated P value to begin a laboratory search for the disease locus performs adequately for a common disease allele. PMID- 11855953 TI - Power calculations for a general class of tests of linkage and association that use nuclear families with affected and unaffected sibs. AB - Family-based tests of association are now often used when trying to fine-map a disease susceptibility locus. Recently, several tests of linkage and association have been proposed that use nuclear families with multiple affected and unaffected sibs rather than just case-parent triads. In this paper we propose a test that generalizes these previous tests. Formulae are derived to calculate the power of the test for a randomly mating population. These power calculations are used to determine conditions under which it is advantageous to include unaffected sibs in the analysis. PMID- 11855954 TI - Detection of human genetic linkage: foundations. AB - Starting with the basics of inheritance, recombination, and genetic map distance, we develop the theory of linkage analysis considering data on an individual or on a sib pair, with and without data on parents. We discuss the effects of ascertainment of individuals with certain traits and the effects of allelic associations between a trait and a linked marker locus. PMID- 11855955 TI - Implications of multilocus inheritance for gene-disease association studies. AB - The impact of multilocus inheritance on the power of candidate gene association studies and the parameters derived therefrom is considered. Both case-control and family-based control designs are included. When the background heritability H (i.e., residual correlation) among sibs for the disease in question is high, the power of multiplex affected sibships versus singletons is diminished compared with the situation when background heritability is low. Thus, multiplex families for association studies are most advantageous for low heritability disorders. Estimates of genotype relative risk (GRR) are also distorted in multiplex sibships compared with singletons based on background heritability. In a case control study, GRR is inflated in multiplex families, with the degree of inflation decreasing with background heritability. By contrast, in a TDT (parental control) study, GRR is deflated in multiplex families, with the degree of deflation increasing with background heritability (and no deflation when H is 0). Considering identical twins, the difference in genotype frequencies between concordant versus discordant MZ twin pairs for the candidate gene is large when the heritability is low, but modest when the heritability is high. PMID- 11855956 TI - Computational issues in mapping variation affecting susceptibility to complex disorders: the chicken and the egg. AB - Linkage mapping strategies for complex disorders have evolved under a variety of constraints. Some of these constraints reflect the nature of complex disorders and are manifest in limitations on the kinds of data that can be collected, while others were (at least historically) strictly computational. This paper focuses on how computational issues have impacted the design of studies on complex disorders and, conversely, how our study designs have influenced the computational issues that have been addressed. We now have unprecedented computational resources, but also face unprecedented computational and methodological challenges as we move from the linkage mapping of genes influencing susceptibility to complex disorders toward the identification of the actual variation affecting susceptibility to these disorders. The near-term computational and methodological issues we must address will be profoundly influenced by the study designs of the recent past. But future study designs, as well as our investments in computational and methodological research, ought to be developed considering the computational and informatics resources we now have at hand. PMID- 11855957 TI - Case-control studies of association in structured or admixed populations. AB - Case-control tests for association are an important tool for mapping complex trait genes. But population structure can invalidate this approach, leading to apparent associations at markers that are unlinked to disease loci. Family-based tests of association can avoid this problem, but such studies are often more expensive and in some cases--particularly for late-onset diseases--are impractical. In this review article we describe a series of approaches published over the past 2 years which use multilocus genotype data to enable valid case control tests of association, even in the presence of population structure. These tests can be classified into two categories. "Genomic control" methods use the independent marker loci to adjust the distribution of a standard test statistic, while "structured association" methods infer the details of population structure en route to testing for association. We discuss the statistical issues involved in the different approaches and present results from simulations comparing the relative performance of the methods under a range of models. PMID- 11855958 TI - Joint tests of linkage and association for quantitative traits. AB - In this paper we describe various study designs and analytic techniques for testing the joint hypothesis that a genetic marker is both linked to and associated with a quantitative phenotype. Issues of power and sampling are addressed. The distinction between methods that explicitly examine association and those that infer association by examining the distribution of allelic transmissions from a heterozygous parent is examined. Extensions to multivariate, multiallelic, and multilocus situations are addressed. Recent approaches that combine variance-components-based linkage analyses with joint tests of linkage in the presence of association for disentanglement of the linkage and association and the application of such methods to fine mapping are discussed. Finally, new classes of joint tests of linkage and association that do not require samples of related individuals are described. PMID- 11855959 TI - Association studies under general disease models. AB - There is great expectation that the levels of association found between genetic markers and disease status will play a role in the location of disease genes. This expectation follows from regarding association as being proportional to linkage disequilibrium and therefore inversely related to recombination value. For disease genes with more than two alleles, the association measure is instead a weighted average of linkage disequilibria, with the weights depending on allele frequencies and genotype susceptibilities at the disease loci. There is no longer a simple relationship, even in expectation, with recombination. We adopt a general framework to examine association mapping methods which helps to clarify the nature of case-control and transmission/disequilibrium-type tests and reveals the relationship between measures of association and coefficients of linkage disequilibrium. In particular, we can show the consequences of additive and nonadditive effects at the trait locus on the behavior of these tests. These concepts have a natural extension to marker haplotypes. The association of two locus marker haplotypes with disease phenotype depends on a weighted average of three-locus disequilibria (two markers with each disease locus). It is likely that these two-marker analyses will provide additional information in association mapping studies. PMID- 11855960 TI - Global molecular identification from graphs. Neutral and ionized main-group diatomic molecules. AB - Diophantine equations and inequalities are presented for main-group closed-shell diatomic molecules. Specifying various bond types (covalent, dative, ionic, van der Waals) and multiplicities, it becomes possible to identify all possible molecules. While many of the identified species are probably unstable under normal conditions, they are interesting and present a challenge for computational or experimental analysis. Ionized molecules with net charges of -1, 1, and 2 are also identified. The analysis applies to molecules with atoms from periods 2 and 3 but can be generalized by substituting isovalent atoms. When closed-shell neutral diatomics are positioned in the chemical space (with axes enumerating the numbers of valence electrons of the free atoms), it is seen that they lie on a few parallel isoelectronic series. PMID- 11855961 TI - Computing wiener-type indices for virtual combinatorial libraries generated from heteroatom-containing building blocks. AB - The expensive and time-consuming process of drug lead discovery is significantly accelerated by efficiently screening molecular libraries with a high structural diversity and selecting subsets of molecules according to their similarity toward specific collections of active compounds. To characterize the molecular similarity/diversity or to quantify the drug-like character of compounds the process of screening virtual and synthetic combinatorial libraries uses various classes of structural descriptors, such as structure keys, fingerprints, graph invariants, and various topological indices computed from atomic connectivities or graph distances. In this paper we present efficient algorithms for the computation of several distance-based topological indices of a molecular graph from the distance invariants of its subgraphs. The procedures utilize vertex- and edge-weighted molecular graphs representing organic compounds containing heteroatoms and multiple bonds. These equations offer an effective way to compute for weighted molecular graphs the Wiener index, even/odd Wiener index, and resistance-distance index. The proposed algorithms are especially efficient in computing distance-based structural descriptors in combinatorial libraries without actually generating the compounds, because only distance-based indices of the building blocks are needed to generate the topological indices of any compound assembled from the building blocks. PMID- 11855962 TI - Electronic Chemistry Conferences: 7 years of CONFCHEM. AB - Between the summer of 1993 and August 2001, members of the ACS Division of Chemical Education's Committee on Computers in Chemical Education (CCCE) managed the online facilities for 17 online conferences. One more conference will occur during fall 2001, and several additional conferences are planned for the future. This article describes the history and the nature of these online conferences, including the advantages and disadvantages of the online format. PMID- 11855963 TI - An alignment-independent versatile structure descriptor for QSAR and QSPR based on the distribution of molecular features. AB - A molecular descriptor based upon a count statistic of the topological distance matrix is described and evaluated for use in QSAR studies. Encoding a molecule is done by computing many selective count statistics (histograms) reflecting the distribution of different atom types and bond types in the molecule. The descriptor was also extended to incorporate geometric features of molecules by weighting the topological distance counts with the geometric distance. It is invariant to both translation and rotation. As a result, it does not require the alignment of the structures under study. The method was applied to several QSAR data sets and performed equally well or better than CoMFA and the EVA descriptor. Compared to the latter two methods, it is computationally easier. PMID- 11855964 TI - Neural networks evaluating NMR data: an approach to visualize similarities and relationships of sol-gel derived inorganic-organic and organometallic hybrid polymers. AB - An artificial neural network (ANN)--the Kohonen Self-Organizing Feature Map (SOM) is used to evaluate solid-state NMR spectroscopic derived data of 72 siloxane based phosphine and organometallic functionalized hybrid polymers. The data set consists of parameters that describe their structural features and their dynamic behavior. The ANN visualizes similarities of the investigated compounds by reducing the dimension of the data set. This allows a comparison of these polymers that was not possible beforehand because of their structural diversity. PMID- 11855965 TI - Enhanced CACTVS browser of the Open NCI Database. AB - A Web-based, graphical user interface has been developed to conduct rapid searches by numerous criteria in the more than 250,000 structures of the Open NCI Database. It is based on the chemistry information toolkit CACTVS. Nearly all structures and anticancer and anti-HIV screening data provided by NCI's Developmental Therapeutics Program have been included. This data set has been augmented by a large amount of additional, mostly computed, data, such as calculated log P values, predicted biological activities, systematically determined names, and others. Complex boolean searches are possible. Flexible substructure searches have been implemented. The user can conduct 3D pharmacophore queries in up to 25 conformations precalculated for each compound. Numerous output formats as well as 2D and 3D visualization options are provided. It is possible to export search results in various forms and with choices for data contents in the exported files, for structure sets ranging in size from a single compound to the entire database. Only a Web browser is needed to use this service, with a few plug-ins being useful but optional. PMID- 11855966 TI - Rapid evaluation of molecular shape similarity index using pairwise calculation of the nearest atomic distances. AB - Rapid evaluation method for obtaining molecular shape similarity index using pairwise calculation of the nearest atomic distance respected to the template atoms was investigated. This method for calculations of similarity indices remarkably reduced required time compared with hitherto methods (especially 2 or 3 orders of magnitude faster than the previous grid-based evaluation technique) and gave without clear loss of preciseness. The potential of these improvements and possible further enhancements are discussed. PMID- 11855967 TI - A new method for fast and accurate derivation of molecular conformations. AB - During molecular simulations, three-dimensional conformations of biomolecules are calculated from the values of their bond angles, bond lengths, and torsional angles. In this paper we study how to efficiently derive three-dimensional molecular conformations from the values of torsional angles. This case is of broad interest as torsional angles greatly affect molecular shape and are always taken into account during simulations. We first review two widely used methods for deriving molecular conformations, the simple rotations scheme and the Denavit Hartenberg local frames method. We discuss their disadvantages which include extensive bookkeeping, accumulation of numerical errors, and redundancies in the local frames used. Then we introduce a new, fast, and accurate method called the atomgroup local frames method. This new method not only eliminates the disadvantages of earlier approaches but also provides lazy evaluation of atom positions and reduces the computational cost. Our method is especially useful in applications where many conformations are generated or updated such as in energy minimization and conformational search. PMID- 11855968 TI - QSPR correlation of the melting point for pyridinium bromides, potential ionic liquids. AB - In an attempt to develop predictive tools for the determination of new ionic liquid solvents, QSPR models for the melting points of 126 structurally diverse pyridinium bromides in the temperature range 30-200 degrees C were developed with the CODESSA program. Six- and two-descriptor equations with squared correlation coefficients (R(2)) of 0.788 and 0.713, respectively, are reported for the melting temperatures. The models illustrate the importance of information content indices, total entropy, and the average nucleophilic reactivity index for an N atom. PMID- 11855969 TI - Prediction of glass transition temperature (T(g)) of some compounds in organic electroluminescent devices with their molecular properties. AB - We have studied the quantitative structure-property relationship between descriptors representing the molecular structure and glass transition temperature (T(g)) for 103 molecules including organic electroluminescent (EL) devices materials. Eighty-six descriptors were introduced and among them seven descriptors (one topological descriptor, one thermodynamic descriptor, one spatial descriptor, one structural descriptor, and three electrostatic descriptors) were selected by Genetic Algorithm (GA). The 81 molecules chosen randomly among 103 compounds were used as a training set, and the remaining 22 molecules were used as a prediction set. The quantitative relationship between these seven descriptors and T(g) was tested by multiple linear regression (MLR) and artificial neural network (ANN). ANN analysis showed no significant advantage over MLR for this study. As the results of the MLR, the square of the correlation coefficient (R(2)) for the T(g) of the 81 training set was 0.989, and the average error was 8.8 K. In prediction for T(g) using the 22 prediction compounds set with MLR, R(2) was 0.976, and the average error was 13.9 K. PMID- 11855970 TI - Spectral moments of polycyclic aromatic hydrocarbons. Solution of a kinetic problem. AB - The relationship between the rate of supercritical fluid extraction of polycyclic aromatic hydrocarbons (PAHs) from coal-tar pitch and some topological invariants are examined. The aim is to explain the appearance of a minimum value on the activation energy/molar mass curve of the PAHs. Rate constants are approximated using spectral moment expansion. It is shown that the size of the PAHs determines more than 95% of the extraction rate. Activation energy for the extraction of the PAHs was found to increase with increasing molar mass. The appearance of a minimum value on the activation energy/molar mass curve is the consequence of experimental difficulties resulting from the tendency of lower members of the PAHs to sublime. PMID- 11855971 TI - Chemical descriptors with distinct levels of information content and varying sensitivity to differences between selected compound databases identified by SE DSE analysis. AB - Analysis of the variability of molecular descriptors in large compound databases has recently been carried out using both the Shannon entropy (SE) and differential Shannon entropy (DSE) concepts that reduce descriptor distributions to their information content (SE analysis) and detect intrinsic differences between descriptor settings in compound databases (DSE analysis). Here it is shown that a combination of SE and DSE calculations, termed SE-DSE analysis, makes it possible to identify molecular descriptors most sensitive to systematic differences in databases consisting of synthetic, drug-like, and natural molecules. Descriptors with consistently high information content are detected, and database-specific differences are quantified. Different sets of only very few descriptors were found to be most responsive to principal differences between synthetic, natural, and drug-like molecules. Descriptors with DSE values furthest away from zero are likely to best distinguish between compounds with different characteristics. SE-DSE analysis also reveals that a number of descriptors are not sensitive to compound class-specific features, despite their complexity and consistently high information content. PMID- 11855972 TI - Development of quantitative structure-activity relationship and classification models for a set of carbonic anhydrase inhibitors. AB - Mathematical models are developed to find quantitative structure-activity relationships that correlate chemical structure and inhibition toward three carbonic anhydrase (CA) isozymes: CA I, II, and IV. Numerical descriptors are generated to encode important topological, geometric, and electronic features of molecular structure. After descriptor generation, multiple linear regression, and computational neural network (CNN) analyses are performed on various descriptor subsets to find superior models for prediction. Committees of five CNNs were utilized to average final predicted values for the 142-compound data set. For inhibitors of CA I, an 8-5-1 CNN committee produced a training set rms error of 0.105 log K(i) (r(2) = 0.994) and prediction set rms error of 0.208 log K(i) (r(2) = 0.980). Training and prediction set rms errors of 0.140 log K(i) (r(2) = 0.992) and 0.231 log K(i) (r(2) = 0.971), respectively, were produced by a 9-5-1 CNN committee for inhibitors of CA II. For prediction of CA IV inhibitors, an 8-5 1 CNN committee produced training and prediction set rms errors of 0.147 log K(i) (r(2) = 0.992) and 0.211 log K(i) (r(2) = 0.991), respectively. In addition, classification models were built using k-nearest neighbor (kNN) analysis to solve two- and three-class problems for inhibitors of CA IV. A three-descriptor classification model proved superior in labeling compounds as active or inactive inhibitors for the two-class problem. Training and prediction set percent classification rates of 100% and 87.1%, respectively, were obtained. For the three-class (active/moderate/inactive) problem, a five-descriptor model was deemed optimal producing a training set percent classification rate of 98.8% and prediction set rate of 79.0%. PMID- 11855973 TI - The most common chemical replacements in drug-like compounds. AB - We have written a method that extracts one-to-one replacements of chemical groups in pairs of drug-like molecules with the same biological activity and counts the frequency of the replacements in a large collection of such molecules. There are two variations on the method that differ in their treatment of replacements in rings. This method is one possible approach to systematically identify candidate bioisosteres. Here we look at the MDDR database because it has a large diversity of drug-like compounds in a large number of therapeutic areas. The most frequent replacements in MDDR seem generally consistent with medicinal chemistry intuition about what chemical groups are equivalent or with groups that are easily converted by synthetic or metabolic pathways. This method can be applied to any set of molecules wherein the molecules can be paired by similar biological activity. PMID- 11855974 TI - Three-dimensional quantitative structure--property relationship (3D-QSPR) models for prediction of thermodynamic properties of polychlorinated biphenyls (PCBs): enthalpy of sublimation. AB - Three-dimensional quantitative structure--property relationship (3D-QSPR) models have been constructed using comparative molecular field analysis (CoMFA) to correlate the sublimation enthalpies at 298.15 K of a series of polychlorinated biphenyls (PCBs) with their CoMFA-calculated physicochemical properties. Various alignment schemes, such as atom fit, as is, and inertial were employed in this study. Separate CoMFA models were developed using different partial charge formalisms, namely, electrostatic potential (ESP) and Gasteiger-Marsili (GM) charges. Among the four different CoMFA models constructed for sublimation enthalpy (Delta(sub)H(m)(298.15 K)), the model that combined atom fit alignment and ESP charges yielded the greatest self-consistency (r(2) = 0.976) and internal predictive ability (r(cv)(2) = 0.750). This CoMFA model was used to predict Delta(sub)H(m)(298.15 K) of PCBs for which the corresponding experimental values are unavailable in the literature. PMID- 11855975 TI - A fractal approach for selecting an appropriate bin size for cell-based diversity estimation. AB - A novel approach for selecting an appropriate bin size for cell-based diversity assessment is presented. The method measures the sensitivity of the diversity index as a function of grid resolution, using a box-counting algorithm that is reminiscent of those used in fractal analysis. It is shown that the relative variance of the diversity score (sum of squared cell occupancies) of several commonly used molecular descriptor sets exhibits a bell-shaped distribution, whose exact characteristics depend on the distribution of the data set, the number of points considered, and the dimensionality of the feature space. The peak of this distribution represents the optimal bin size for a given data set and sample size. Although box counting can be performed in an algorithmically efficient manner, the ability of cell-based methods to distinguish between subsets of different spread falls sharply with dimensionality, and the method becomes useless beyond a few dimensions. PMID- 11855977 TI - EndNote 5 for windows. PMID- 11855976 TI - Prediction of beta-turns in proteins using the first-order Markov models. AB - We present a method based on the first-order Markov models for predicting simple beta-turns and loops containing multiple turns in proteins. Sequences of 338 proteins in a database are divided using the published turn criteria into the following three regions, namely, the turn, the boundary, and the nonturn ones. A transition probability matrix is constructed for either the turn or the nonturn region using the weighted transition probabilities computed for dipeptides identified from each region. There are two such matrices constructed for the boundary region since the transition probabilities for dipeptides immediately preceding or following a turn are different. The window used for scanning a protein sequence from amino (N-) to carboxyl (C-) terminal is a hexapeptide since the transition probability computed for a turn tetrapeptide is capped at both the N- and C- termini with a boundary transition probability indexed respectively from the two boundary transition matrices. A sum of the averaged product of the transition probabilities of all the hexapeptides involving each residue is computed. This is then weighted with a probability computed from assuming that all the hexapeptides are from the nonturn region to give the final prediction quantity. Both simple beta-turns and loops containing multiple turns in a protein are then identified by the rising of the prediction quantity computed. The performance of the prediction scheme or the percentage (%) of correct prediction is evaluated through computation of Matthews correlation coefficients for each protein predicted. It is found that the prediction method is capable of giving prediction results with better correlation between the percent of correct prediction and the Matthews correlation coefficients for a group of test proteins as compared with those predicted using some secondary structural prediction methods. The prediction accuracy for about 40% of proteins in the database or 50% of proteins in the test set is better than 70%. Such a percentage for the test set is reduced to 30 if the structures of all the proteins in the set are treated as unknown. PMID- 11855978 TI - Rhodacyanine dyes as antimalarials. 1. Preliminary evaluation of their activity and toxicity. AB - The rhodacyanine dye MKT-077 (1), a potent antitumor agent, was found to possess strong in vitro activity against Plasmodium falciparum and a low cytotoxicity. Several new rhodacyanine dyes related to 1, containing a variety of linked heterocyclic moieties, were synthesized, and their antimalarial potencies were evaluated. The synthetic rhodacyanines were found to have EC(50) values against P. falciparum in vitro in the range of 4-300 nM. Several compounds in this series have remarkable selective toxicity profiles (>100). PMID- 11855979 TI - Identification of novel inhibitors of the transforming growth factor beta1 (TGF beta1) type 1 receptor (ALK5). AB - Screening of our internal compound collection for inhibitors of the transforming growth factor beta1 (TGF-beta1) type I receptor (ALK5) identified several hits. Optimization of the dihydropyrroloimidazole hit 2 by introduction of a 2-pyridine and 3,4-methylenedioxyphenyl group gave 7, a selective ALK5 inhibitor. With this information, optimization of the triarylimidazole hit 8 gave the selective inhibitor 14, which inhibits TGF-beta1-induced fibronectin mRNA formation while displaying no measurable cytotoxicity in the 48 h XTT assay. PMID- 11855980 TI - Bis(1H-2-indolyl)methanones as a novel class of inhibitors of the platelet derived growth factor receptor kinase. AB - The novel lead bis(1H-2-indolyl)methanone inhibits autophosphorylation of platelet-derived growth factor (PDGF) receptor tyrosine kinase in intact cells. Various substituents in the 5- or 6-position of one indole ring increase or preserve potency, whereas most modifications of the ring structures and of the methanone group as well as substitution at both indoles result in weak or no activity. An ATP binding site model, derived by homology from the FGFR-1 tyrosine kinase crystal structure suggesting hydrogen bonds of one indole NH and the methanone oxygen with the backbone carbonyl and amide, respectively, of Cys684, explains why only one indole moiety is open for substitution and locates groups in the 5- or 6-position outside the pocket. The hitherto most active derivatives, 39, 53 and 67, inhibit both isoforms of the PDGF receptor kinase in intact cells, with IC(50) of 0.1-0.3 microM, and purified PDGFbeta-receptor in vitro, with IC(50) of 0.09, 0.1, or 0.02 microM, respectively. PDGF-stimulated DNA synthesis is inhibited by these derivatives with IC(50) values of 1-3 microM. Kinetic analysis of 53 showed an ATP-competitive mode of inhibition. The compounds are inactive or weakly active toward a number of other tyrosine kinases, including the FGF receptor 1, EGF receptor, and c-Src kinase, as well as toward serine threonine kinases, including different PKC isoforms and GRK2, and appear therefore selective for PDGF receptor inhibition. PMID- 11855981 TI - Solution structure of a putative HIV1 immunogenic peptide: computer simulation of the principal CD4 binding domain of gp120. AB - A 15-residue segment within the principal CD4 binding domain of gp120 from HIV1 is a natural subject for vaccine development. Knowledge of a three-dimensional structure of this peptide in aqueous solution can in principle be used for pharmacophoric footprint identification. However, the peptide is resistant to structural characterization using nuclear magnetic resonance (NMR) due to aggregation properties. Here we examine the conformational properties of the peptide using molecular dynamics simulation. Three simulations were performed starting from widely different structures of the peptide determined by NMR in organic solvents. The three structures converge during the simulations to a common structure in both Cartesian and in backbone dihedral conformational space. Preliminary examination of the average three-dimensional structure identifies, after sequence threading, characteristics of potential use as a pharmacophoric footprint. PMID- 11855982 TI - Secondary structure of the third extracellular loop responsible for ligand selectivity of a mammalian gonadotropin-releasing hormone receptor. AB - The extracellular loop 3 (ECL3) of the mammalian gonadotropin-releasing hormone receptor (GnRH-R) contains an acidic amino acid (Glu(301) in the mouse GnRH-R) that confers agonist selectivity for Arg(8) in mammalian GnRH. It is proposed that a specific conformation of ECL3 is necessary to orientate the carboxyl side chain of the acidic residue for interaction with Arg(8) of GnRH, which is supported by decreased affinity for Arg(8) GnRH but not Gln(8) GnRH when an adjacent Pro is mutated to Ala. To probe the structural contribution of the loop domain to the proposed presentation of the carboxyl side chain, we synthesized a model peptide (CGPEMLNRVSEPGC) representing residues 293-302 of mouse ECL3, where Cys and Gly residues are added symmetrically at the N and C termini, respectively, allowing the introduction of a disulfide bridge to simulate the distances at which the ECL3 is tethered to the transmembrane domains 6 and 7 of the receptor. The ability of the ECL3 peptide to bind GnRH with low affinity was demonstrated by its inhibition of GnRH stimulation of inositol phosphate production in cells expressing the GnRH-R. The CD bands of the ECL3 peptides exhibited a superposition of predominantly unordered structure and partial contributions from beta-sheet structure. Likewise, the analysis of the amide I and amide III bands from micro-Raman and FT Raman experiments revealed mainly unordered conformations of the cyclic and of the linear peptide. NMR data demonstrated the presence of a beta-hairpin among an ensemble of largely disordered structures in the cyclic peptide. The location of the turn linking the two strands of the hairpin was assigned to the three central residues L(296), N(297), and R(298). A small population of structured species among an ensemble of predominantly random coil conformation suggests that the unliganded receptor represents a variety of structural conformers, some of which have the potential to make contacts with the ligand. We propose a mechanism of receptor activation whereby binding of the agonist to the inactive receptor state induces and stabilizes a particular structural state of the loop domain, leading to further conformational rearrangements across the transmembrane domain and signal propagating interaction with G proteins. Interaction of the Glu(301) of the receptor with Arg(8) of GnRH induces a folded configuration of the ligand. Our proposal thus suggests that conformational changes of both ligand and receptor result from this interaction. PMID- 11855983 TI - Synthesis and biological evaluation of a new set of pyrazolo[1,5-c]quinazoline-2 carboxylates as novel excitatory amino acid antagonists. AB - In recent papers (Catarzi, D.; et al. J. Med. Chem. 1999, 42, 2478-2484; 2000, 43, 3824-3826; 2001, 44, 3157-3165) we reported the synthesis of a set of 4,5 dihydro-4-oxo-1,2,4-triazolo[1,5-a]quinoxaline-2-carboxylates (TQXs) that were active at the Gly/NMDA and/or AMPA receptors. In the present work the synthesis and Gly/NMDA, AMPA, and KA receptor binding affinities of a set of 5,6-dihydro-5 oxo-pyrazolo[1,5-c]quinazoline-2-carboxylates 1a,b-4a,b, 5a, 6a, and 7a,b-9a,b, (+/-)-5,6-dihydro-pyrazolo[1,5-c]quinazoline-2,5-dicarboxylates 10a,b and 11a,b, and (+/-)-1,5,6,10b-tetrahydro-5-oxo-pyrazolo[1,5-c]quinazoline-2-carboxylates 12a,b-14a,b are reported. The binding results indicate that compounds 1a,b-4a,b, 5a, 6a, and 7a,b-9a,b show good Gly/NMDA and/or AMPA receptor binding affinities, demonstrating that the pyrazoloquinazoline tricyclic system is an adequate alternative to the triazoloquinoxaline framework for anchoring at both receptor types. Moreover, the inactivity of the 2,5-dicarboxylate derivatives 10a,b and 11a,b at the Gly/NMDA and AMPA receptors indicates that the presence of a glycine moiety in the southern portion of the pyrazoloquinazoline framework is deleterious for receptor-ligand interaction. Finally, the binding data of compounds 12a,b-14a,b indicate that lack of planarity in the northeastern region of the molecules shifts selectivity toward the Gly/NMDA receptor, depending on the benzofused substitutions. In general, the pyrazoloquinazoline derivatives herein reported were inactive at the KA receptor. A study of the functional antagonism at both the AMPA receptor and the NMDA receptor-ion channel complex was also performed on some selected compounds. PMID- 11855984 TI - Nanomolar small molecule inhibitors for alphav(beta)6, alphav(beta)5, and alphav(beta)3 integrins. AB - Integrin adhesion receptors frequently recognize a core amino acid sequence, Arg Gly-Asp, in their target ligands. Inhibitors with the ability to inhibit one or a small subset of such RGD-dependent integrins have been invaluable in defining their biological function. Here, we have characterized low molecular weight inhibitors for their ability to specifically inhibit alphav(beta)6 integrin, a fibronectin/tenascin receptor. As of yet, no nonpeptidic inhibitor of alphav(beta)6 was known. New peptidomimetic and nonpeptidic compounds were examined in isolated integrin binding assays and in cell adhesion assays for their ability to block alphav(beta)6, alphav(beta)3, alphav(beta)5, and alphalIb(beta)3 integrins. The compounds are based on an aromatically substituted beta amino acid or glutaric acid derivative as an acidic center and an aminopyridyl or guanidyl residue as a basic mimetic. We found several classes of inhibitors with different selectivities, especially mono- or biselectivity on the alpha(v)-integrins alphav(beta)6 and alphav(beta)3, and nanomolar activity. Furthermore, nearly all compounds are inactive on alphaIIb(beta)3. Compound 11 is the first specific, peptidomimetic inhibitor of the alphav(beta)6 integrin receptor. PMID- 11855985 TI - Mechanism-based design of parasite-targeted artemisinin derivatives: synthesis and antimalarial activity of new diamine containing analogues. AB - The potent antimalarial activity of chloroquine against chloroquine-sensitive strains can be attributed, in part, to its high accumulation in the acidic environment of the heme-rich parasite food vacuole. A key component of this intraparasitic chloroquine accumulation mechanism is a weak base "ion-trapping" effect whereupon the basic drug is concentrated in the acidic food vacuole in its membrane-impermeable diprotonated form. By the incorporation of amino functionality into target artemisinin analogues, we hoped to prepare a new series of analogues that, by virtue of increased accumulation into the ferrous-rich vacuole, would display enhanced antimalarial potency. The initial part of the project focused on the preparation of piperazine-linked analogues (series 1 (7 16)). Antimalarial evaluation of these derivatives demonstrated potent activity versus both chloroquine-sensitive and chloroquine-resistant parasites. On the basis of these observations, we then set about preparing a series of C-10 carba linked amino derivatives. Optimization of the key synthetic step using a newly developed coupling protocol provided a key intermediate, allyldeoxoartemisinin (17) in 90% yield. Further elaboration, in three steps, provided nine target C-10 carba analogues (series 2 (21-29)) in good overall yields. Antimalarial assessment demonstrated that these compounds were 4-fold more potent than artemisinin and about twice as active as artemether in vitro versus chloroquine resistant parasites. On the basis of the products obtained from biomimetic Fe(II) degradation of the C-10 carba analogue (23), we propose that these analogues may have a mode of action subtly different from that of the parent drug artemisinin (series 1 (7-16)) and other C-10 ether derivatives such as artemether. Preliminary in vivo testing by the WHO demonstrated that four of these compounds are active orally at doses of less than 10 mg/kg. Since these analogues are available as water-soluble salts and cannot form dihydroartemisinin by P450 catalyzed oxidation, they represent useful leads that might prove to be superior to the currently used derivatives, artemether and artesunate. PMID- 11855986 TI - Synthesis and nicotinic binding studies on enantiopure diazine analogues of the novel (2-chloro-5-pyridyl)-9-azabicyclo[4.2.1]non-2-ene UB-165. AB - As part of our program aimed at optimizing therapeutic effects over toxic effects (as observed in the naturally occurring nicotinic acetylcholine receptor modulators (-)-nicotine, (-)-epibatidine, (-)-ferruginine, and (+)-anatoxin-a), we investigated the bioisosteric potential of diazines in the field of (+) anatoxin-a-type structures. In the series of diazine analogues of deschloro-UB 165 (DUB-165, 6), bioisosteric replacement of the 3-pyridyl pharmacophoric element by a 4-pyridazinyl, 5-pyrimidinyl, or 2-pyrazinyl moiety resulted in novel nAChR ligands 7, 8, and 9. A palladium-catalyzed Suzuki cross-coupling of the 3-diethylboranylpyridine (14) and a Stille cross-coupling of the corresponding tributylstannyl diazines 15-17 with the vinyl triflate 13 of the N protected 9-azabicyclo[4.2.1]nonan-2-one 12 constitute the key steps in the syntheses of these enantiopure anatoxinoids 6-9. Studies of the in vitro affinity for (alpha4)(2)(beta2)(3), alpha3(beta)4, and alpha7 nAChR subtypes by radioligand binding assays demonstrated that the diazine analogues 7-9 can be considered as pharmacologically attractive bioisosteres of DUB-165 (6) but with different effects on the binding affinity with regard to the diazine moiety. The pyrimidine-containing bioisostere 8 turned out to be the most active diazine analogue, which interacts potently (K(i) = 0.14 nM) with the (alpha4)(2)(beta2)(3) subtype and differentiates significantly among the nAChR subtypes investigated. The nitrogens in this anatoxinoid 8 show by far the most negative atomic charges (calculated using the AM1 Hamiltonian). This qualitatively correlates with the highest binding affinity observed for 8 for all subtypes under consideration. PMID- 11855988 TI - Synthesis and antitumor activity of several new analogues of penclomedine and its metabolites. AB - Analogues of penclomedine (PEN, 3,5-dichloro-4,6-dimethoxy-2 (trichloromethyl)pyridine) and its metabolites have been synthesized and evaluated as potential antitumor agents. PEN and 4-DMPEN (3,5-dichloro-4-hydroxy 6-methoxy2-(trichloromethyl)pyridine (3a)), the major plasma metabolite in patients, were modified at 4- and 6-positions with different alkyl, aryl, and ester groups. All of the analogues and many of the intermediates were evaluated against the PEN-sensitive MX-1 human breast tumor xenograft in vivo, and several analogues of PEN and 4-DMPEN showed modest to curative activity. PMID- 11855987 TI - Synthesis and biolocalization of water-soluble sapphyrins. AB - The synthesis, characterization, and biolocalization properties of new, fully water-soluble sapphyrins is reported. These systems, which display light absorbing features that make them of potential interest as photodynamic therapy (PDT) sensitizers, were found to localize selectively in pancreatic carcinoma tissue in a xenographic murine model. In this model, the concentration ratios for malignant tissue to surrounding muscle tissue were found to be as high as 280 in the most favorable case. The concentrations of the water-soluble sapphyrins reported here, as well as a porphyrin control system, were determined as a function of time and tissue type. PMID- 11855990 TI - Feature selection for structure-activity correlation using binary particle swarms. AB - We present a new feature selection algorithm for structure-activity and structure property correlation based on particle swarms. Particle swarms explore the search space through a population of individuals that adapt by returning stochastically toward previously successful regions, influenced by the success of their neighbors. This method, which was originally intended for searching multidimensional continuous spaces, is adapted to the problem of feature selection by viewing the location vectors of the particles as probabilities and employing roulette wheel selection to construct candidate subsets. The algorithm is applied in the construction of parsimonious quantitative structure-activity relationship (QSAR) models based on feed-forward neural networks and is tested on three classical data sets from the QSAR literature. It is shown that the method compares favorably with simulated annealing and is able to identify a better and more diverse set of solutions given the same amount of simulation time. PMID- 11855989 TI - 6-Sulfonylchromenes as highly potent K(ATP)-channel openers. AB - We synthesized K(ATP)-channel openers (KCOs) composed of the 4-pyridonechromene moiety of bimakalim (1) and a variety of sulfonyl-containing 6-substituents 4-29. Dilator potencies were measured in rat aorta and trachea. In both test systems the KCOs exhibit potency ranges of roughly 3 log units. The 6-N-phenyl-N methylsulfonamido derivative 24 shows the highest potency. In rat aorta the potency spectrum ranges from a pEC(50) value of 8.76 to 5.68; in rat trachea it ranges from 8.01 to 4.99. On average, the dilator activity is about 0.8 log units stronger in the aorta. Aortic relaxation by chromene 13 is markedly retarded, the clinical relevance of which (e.g., preventing tachycardia) remains to be clarified. Binding affinities were determined in myocardial membranes and aortic smooth muscle cells of the rat. The affinity spectrum in myocardial membranes ranges from a pK(D) of 7.83 to 5.18; the highest affinity in aortic smooth muscle cells is measured for compound 28 (pK(D) = 8.55), whereas the lowest affinity is measured for 4 (pK(D) = 4.51). Significant selectivities discriminating between K(ATP)-channels of different organs could not be detected. PLS analysis yielded no significant correlation between vasodilator activity in aorta and chemical descriptors (GRIND). Compounds 13, 24, and 28 represent the most potent KCOs of the 4-pyridonechromene type published so far. Their 6-substituents exhibit a phenyl ring with a congruent conformational orientation in relation to the sulfonylchromene. From SAR data and conformational analysis we postulate that these new 6-substituents extend the binding site for chromene KCOs. Correspondingly, we assume that the receptor area exhibits two separate interaction sites with the capacity to bind 6-substituents: (a) one site interacting with negatively polarized partial structures (e.g., CN, NO(2), SO(2)) and (b) one spatially restricted site enabling favorable pi-interactions. PMID- 11855991 TI - Side-chain lactam-bridge conformational constraints differentiate the activities of salmon and human calcitonins and reveal a new design concept for potent calcitonin analogues. AB - We have recently reported the potent hypocalcemic effects of side-chain lactam bridged analogues of human calcitonin (hCT) (Kapurniotu, A.; et al. Eur. J. Biochem. 1999, 265, 606-618). To extend these studies, we have now synthesized a new series of (Asp(17), Lys(21)) and (Asp(17), Orn(21)) side-chain bridged salmon calcitonin (sCT) and hCT analogues. The affinities of these analogues for the human calcitonin receptor, hCTR(I1)(-), and for rat-brain membrane receptors were assayed in competitive binding assays, and agonist potencies at the hCTR(I1)(-) receptors were assessed, using a cAMP-responsive gene-reporter assay. The bridged sCT analogues had activities similar to sCT itself. In contrast, an (Asp(17), Orn(21)) side-chain bridged hCT analogue, cyclo(17-21)-[Nle(8), Phe(12), Asp(17), Orn,(21) Tyr(22))-hCT, was 80 and 450 times more active than hCT in the hCTR(I1)( ) and rat-brain receptor binding assays, respectively, and was 90 times more potent than hCT and 16 times more potent than sCT in initiating receptor signaling. An uncyclized, isosteric analogue of this peptide was also more potent than hCT, demonstrating that the cyclization constraint and these single-residue substitutions enhance the activities of hCT in an additive fashion. This study demonstrates that the potency-enhancing effects of lactam-bridge constraints at hCT residues 17-21 are not transferable to sCT. We also show that, in comparison to the hCT analogues, sCT and its analogues are less potent agonists than expected from their hCTR(I1)(-) affinities. This suggests that it may be possible to preserve the efficient signal transduction of hCT while introducing additional receptor affinity-enhancing elements from sCT into our potent lactam-bridged hCT analogue, thereby creating new super-potent, hCT-based agonists. PMID- 11855992 TI - Synthesis, pharmacokinetics, efficacy, and rat retinal toxicity of a novel mitomycin C-triamcinolone acetonide conjugate. AB - A novel conjugate of mitomycin C (MMC) and triamcinolone acetonide (TA) was synthesized using glutaric acid as a linker molecule. To determine the rate of hydrolysis, the conjugate was dissolved in aqueous solution and the rate of appearance of free MMC and TA was determined by high-performance liquid chromatography analysis. Antiproliferative activity of the MMC-TA conjugate and parent compounds was assessed using an NIH 3T3 fibroblast cell line. Cell growth was quantified using the MTT assay. Kinetic analysis of the hydrolysis rate demonstrated that the conjugate had a half-life of 23.6 h in aqueous solutions. The antiproliferative activities of the MMC-TA conjugate and MMC were both concentration dependent, with similar IC(50) values of 2.4 and 1.7 microM, respectively. However, individual responses at concentrations above 3 microM showed that the conjugate was less active than MMC alone. TA alone showed only limited inhibition of cell growth. Studies evaluating intravitreal injection of the conjugate demonstrate that this agent produced no measurable toxicity. Our data provide evidence that the MMC-TA conjugate could be used as a slow-release drug delivery system. This could in turn be used to modulate a posttreatment wound healing process or to treat various proliferative diseases. PMID- 11855993 TI - Development of a new class of nonimidazole histamine H(3) receptor ligands with combined inhibitory histamine N-methyltransferase activity. AB - In search of novel ways to enhance histaminergic neurotransmission in the central nervous system, a new class of nonimidazole histamine H(3) receptor ligands were developed that simultaneously possess strong inhibitory activity on the main histamine metabolizing enzyme, histamine N-methyltransferase (HMT). The novel compounds contain an aminoquinoline moiety, which is an important structural feature for HMT inhibitory activity, connected by different spacers to a piperidino group (for H(3) receptor antagonism). Variation of the spacer structure provides two different series of compounds. One series, having only an alkylene spacer between the basic centers, led to highly potent HMT inhibitors with moderate to high affinity at human histamine H(3) receptors. The second series possesses a p-phenoxypropyl spacer, which may be extended by another alkylene chain. This latter series also showed strong inhibitory activity on HMT, and in most cases, the H(3) receptor affinity even surpassed that of the first series. One of the most potent compounds with this dual mode of action is 4-(4-(3 piperidinopropoxy)phenylamino)quinoline (34) (hH(3), K(i) = 0.09 nM; HMT, IC(50) = 51 nM). This class of compounds showed high antagonist potency and good H(3) receptor selectivity in functional assays in guinea pig on H(1), H(2), and H(3) receptors. Because of low or missing in vivo activity of two selected compounds, the proof of concept of these valuable pharmacological tools for the supposed superior overall enhancing effect on histaminergic neurotransmission failed to appear hitherto. PMID- 11855994 TI - New stereocontrolled synthesis and biological evaluation of 5-(1'-hydroxyalkyl)-3 methylidenetetrahydro-2-furanones as potential cytotoxic agents. AB - A series of 3-methylidenetetrahydro-2-furanones 7 bearing various hydroxyalkyl substituents in position 5 were synthesized using novel diastereo- and enantioselective methodology. In vitro cytotoxicity data demonstrated that all prepared compounds were active against L-1210 and HL-60 tissue culture cells with 7e being the most potent (IC(50) = 6.9 microM). Also an increase in activity with an increase in lipophilicity of the substituents in the order H < alkyl < phenyl was observed. PMID- 11855997 TI - Alkylating capacity and reaction products of antimalarial trioxanes after activation by a heme model. AB - The reactivity of 1,2,4-trioxane molecules 2-5, structurally related to the antimalarial drug artemisinin, with a heme model, manganese(II) tetraphenylporphyrin, is reported. With the pharmacologically active drugs 2-4, covalent adducts were obtained by addition of a drug-derived radical onto the porphyrin macrocycle, whereas no reaction was obtained with the nonactive compound 5. This confirms that alkylation is probably one of the key factors of the pharmacological activity of endoperoxide-based antimalarial drugs. PMID- 11855995 TI - Synthesis and biological evaluation of a new furo[2,3-h]quinolin-2(1H)-one. AB - A new furoquinolinone derivative, namely 4-hydroxymethyl-1,6,8-trimethylfuro[2,3 h]quinolin-2(1H)-one (HOFQ), was synthesized and its biological activity studied. By UVA activation, HOFQ induced strong antiproliferative effects in Ehrlich ascite cells, which lost their ability to transmit the tumor by transplantation. HOFQ exhibited poor genotoxicity and absence of skin phototoxicity. Actually, HOFQ sensitization forms DNA-protein cross-linkages but not interstrands cross links. Therefore, HOFQ appears to be a new promising drug for PUVA photochemotherapy and photopheresis. PMID- 11855998 TI - Total synthesis of isoroquefortine C. AB - A short and efficient total synthesis of isoroquefortine C, the 3,12-(Z)-isomer of roquefortine C, from L-tryptophan methyl ester hydrochloride and 4(5) (hydroxy)methylimidazole hydrochloride is described. PMID- 11855999 TI - Theoretical study on chlorine and hydrogen shift in cycloheptatriene and cyclopentadiene derivatives. AB - The transition structures (TSs) for chlorine 1,7-shift and 1,5-shift in 1,7,7 trichlorocycloheptatriene (1) and those of chlorine 1,5-shifts in 1,5,5 trichlorocyclopentadiene (3) and 1,2,5-trichloro-1,3-pentadiene (5) derivatives have been located with density functional theory (DFT) at the Becke3LYP/6-311G [and Becke3LYP/6-311+G] level. The calculational results were compared with those for corresponding hydrogen shifts in nonsubstituted molecules (cycloheptatriene (2), cyclopentadiene (4), and 1,3-pentadiene (6)). The following points were clarified: (1) The activation energy (Delta E(++)) for chlorine 1,7-shift in 1 was evaluated to be only +50.1 [+49.2] kJ/mol, which is smaller than that (+69.9 [+68.3]) for a 1,5-shift, supporting the theory that the conversion between two equivalent A and A' proceeds through a TS for direct chlorine 1,7-shift (Figure 1), rather than through a TS for a 1,5-shift (Figure 2). (2) The considerable amount of charge separation between a migrating chlorine atom (Cl(m)) and a seven membered ring (-0.53 and +0.47 for Merz-Singh-Kollman scheme) occurs in a chlorine 1,7-shift, which is in good contrast to the result that the migrating hydrogen atom (H(m)) for a 1,7-shift in cycloheptatriene (2) carries almost no charge (Figure 3). This large charge separation can stabilize the TS for the chlorine 1,7-shift pathway. (3) The Delta E(++) values for suprafacial hydrogen 1,7-shift in 2 are quite large (+288.0 [+284.8] kJ/mol), much larger than that (+166.8 [+167.0]) for a 1,5-shift in 4 which is orbital symmetrically allowed (Figure 3). The calculation suggests that the chlorine 1,7-shift in 1 occurs easily at room temperature (actually observed experimentally) by proceeding via concerted suprafacial 1,7-shift through the zwitterionic TS with the significant assistance of Coulomb interaction between charged fragments (negatively charged chlorine atom and positively charged tropylium ring), rather than via a suprafacial 1,5-sigmatropic pathway. Other cases studied in this paper showed usual results predicted by orbital symmetrical consideration. PMID- 11856000 TI - Peptide design using omega-amino acids: unusual turn structures nucleated by an N terminal single gamma-aminobutyric acid residue in short model peptides. AB - Incorporation of omega-amino acids into peptide sequences plays an important role in designing peptides with modified backbone conformation and enhanced stability against proteolysis. The present study establishes the presence of unusual turns involving 12-membered hydrogen bonded rings in terminally blocked tri- and tetrapeptides. X-ray diffraction analysis of single crystals and NMR studies have been used to probe the three-dimensional structures of two terminally protected short peptides, Boc-gamma-Abu(1)-Aib(2)-Ala(3)-OMe 1 and Boc-gamma-Abu(1)-Aib(2) Ala(3)-Aib(4)-OMe 2 (gamma-Abu = gamma-aminobutyric acid), in which conformationally flexible omega-amino acids (gamma-Abu) and conformationally restricted alpha-aminoisobutyric acid (Aib) residues are positioned contiguously. The crystal structures of both peptides 1 and 2 exhibit unusual turns composed of 12-membered hydrogen bonded rings involving C [double bond] O from the Boc-group and Ala(3) NH. A type I' beta-turn was observed in the structure of peptide 2 adjacent to the unusual turn with a hydrogen bond between gamma-Abu(1) C [double bond] O and Aib(4) NH. The crystals of peptide 1 are in the space group P2(1), a = 9.3020(10) A, b = 23.785(2) A, c = 10.022(3) A, beta = 101.35 degrees(4), Z = 4, R = 5.7%, and R(w) = 14.5%. Similarly, the crystals of peptide 2 are in the space group C2, a = 19.0772(6) A, b = 8.7883(2) A, c = 16.7758(3) A, beta = 110.7910 degrees(10), Z = 4, R = 6.71%, and R(w) = 15.11%. The unusual turn in both peptides 1 and 2 are retained in solution as is evident from NMR studies in CDCl(3). The role of the adjacently located Aib residue to nucleate the 12 membered hydrogen bonded ring is also addressed. PMID- 11856001 TI - Preparation of optically pure propargylic and allylic alcohols from 2 (trimethylsilyl)vinyl sulfoxides as a chiral ethynyl anion synthon: computational studies on elimination reaction of 2-(trimethylsilyl)vinyl sulfoxides. AB - The reaction of the alpha-carbanion derived from (trimethylsilyl)vinyl sulfoxides with aldehydes afforded a diastereomeric mixture of the products. Each diastereomer was subjected to specific elimination reactions to give optically pure propargylic, trimethylsilylated propargylic, and allylic alcohols. Acceleration of the sulfenic acid-elimination from the beta-silylvinyl sulfoxide was demonstrated by the ab initio calculation to be ascribed mainly to the beta effect of the silyl group. PMID- 11856002 TI - Stereoselective formal [3 + 2] cycloaddition of N-alkylidene glycine ester anions to chiral Fischer alkenylcarbene complexes. Asymmetric synthesis of 3,4,5 trisubstituted prolines. AB - The reaction between N-alkylidene glycine ester enolates, generated from glycine esters aldimines with LDA in THF at low temperature, and chiral alkoxyalkenylcarbene complexes of chromium provided directly 2,4,5-trisubstituted 3-pyrrolidinylcarbene complexes with total exo selectivity and very high syn and facial diastereoselectivity when carbene complexes bearing the (-)-8 phenylmenthyloxy group were employed. Oxidation of the metal carbene moiety followed by basic hydrolysis of the esters afforded enantiomerically highly enriched syn,exo-3,4,5-trisubstituted prolines, whereas acidic hydrolysis of the same functional groups proceeded with epimerization at the alpha-amino acid center leading to anti,exo-3,4,5-trisubstituted prolines of very high enantiomeric purity as well. PMID- 11856003 TI - Nickel-catalyzed intermolecular coupling of 1,3-dienes and aldehydes via transmetalation of nickelacycles with diisobutylaluminum acetylacetonate. AB - Intermolecular coupling reactions of 1,3-dienes and aldehydes via transmetalation of nickelacycle intermediate with (i)Bu(2)Al-acac were investigated. In the reactions, a linear adduct or a branched adduct was produced, depending upon the nature of 1,3-dienes and aldehydes, via two nickelacycles that were relatively stable among the four possible nickelacycles because of the equilibrium with pi allynickel forms. PMID- 11856004 TI - Structure of 9,10-di(9,9-dioctylfluorenyl)anthracene in the solid and in solution. AB - A new example of magnetic nonequivalence of chemically equivalent atoms is identified from the proton and carbon resonance spectra of 9,10-di(9,9 dioctylfluorenyl)anthracene with the aid of its conformation in the crystalline state. Molecular modeling suggests that it has a similar conformation in solution. PMID- 11856005 TI - Novel photodegradation of the antifungal antibiotic pyrrolnitrin in anhydrous and aqueous aprotic solvents. AB - The UV irradiation of pyrrolnitrin (1a), which is an antibiotic clinically useful against dermatophytosis and possesses a unique 2-(pyrrol-3-yl)nitrobenzene moiety in the molecule, in an anhydrous aprotic solvent resulted in the exclusive formation of transient 7,4'-dichlorospiro[1,3-dihydrobenzo(c)isoxazole-3,3' pyrrolin-2'-one] (2a) via the intramolecular oxidation of the juxtaposed pyrrole ring by the triplet-excited nitro group. The irradiation in an aqueous aprotic solvent, however, allowed the concurrent occurrence of intramolecular cyclization by the singlet-excited nitro group in 1a and the hydroxylation at the 2-position of the pyrrole ring by water to afford 3,7-dichloro-8-hydroxy-8,8a dihydropyrrolo[2,3-b]indol-2-one (3a), accompanied by the formation of 2a. Elongation of the irradiation time in these photoreactions caused a rapid consumption of the products, 2a and 3a, to give undetermined polar polymeric products. The present results indicate that the photodegradation of 1a is significantly influenced by the presence of water in the reaction media and by the nature of its excited state. Thus, the loss of the antifungal activities by the photosensitive antibiotic 1a was chemically proved. PMID- 11856007 TI - Shape recognition of alkylammonium ions by 1,3-bridged calix[5]arene crown-6 ethers: endo- vs exo-cavity complexation. AB - A series of tri-O-substituted 1,3-bridged calix[5]arene crown-6 ethers bearing alkyl, arylalkyl, alkoxyalkyl, and alkoxycarbonylmethyl residues at the lower rim and either (t)()Bu or H substituents at the upper rim have been synthesized. (1)H NMR studies have shown that p-tert-butylcalix[5]crowns, irrespective of the size and nature of their lower rim pendant groups, adopt preorganized conelike conformations, whereas p-H-calix[5]crowns with bulky substituents preferentially exist in solution as partial cone conformers (C(1) symmetry). Calix[5]crown derivatives behave as mono- or ditopic receptors for isomeric butylammonium ions, forming endo-cavity (inside the calixarene cup) and/or exo-cavity (at the crown ether moiety) 1:1 complexes according to the shape of the guest. These two binding modes can be clearly distinguished and monitored by (1)H NMR titration experiments. PMID- 11856006 TI - Cesium effect: high chemoselectivity in direct N-alkylation of amines. AB - A novel method for the mono-N-alkylation of primary amines, diamines, and polyamines was developed using cesium bases in order to prepare secondary amines efficiently. A cesium base not only promoted alkylation of primary amines but also suppressed overalkylations of the produced secondary amines. Various amines, alkyl bromides, and alkyl sulfonates were examined, and the results demonstrated this methodology was highly chemoselective to favor mono-N-alkylation over dialkylation. In particular, use of either sterically demanding substrates or amino acid derivatives afforded the secondary amines exclusively, offering wide applications in peptidomimetic syntheses. PMID- 11856008 TI - Photosolvolysis of (E)-styryl(phenyl)iodonium tetrafluoroborate. Generation and reactivity of a primary vinyl Cation. AB - The photochemistry of (E)-styryl(phenyl)iodonium tetrafluoroborate in methanol and 2,2,2-trifluoroethanol as well as in dichloromethane and toluene has been investigated. In all solvents the vinylic C [bond] I bond is more photoreactive than the aromatic C [bond] I bond. Homolysis as well as heterolysis of both bonds occurs, but the latter type of cleavage predominates. In alcoholic solvents, the incipient phenyl cation produces a nucleophilic substitution product. The primary styryl cation gives nucleophilic substitution, elimination, and rearrangement products. The dependence of the photoreaction on the nucleophilicity of the solvent indicates that in the presence of good nucleophiles a 10-I-3 compound is the reactive iodonium species. In this case the reaction proceeds via an S(N)i mechanism. In the absence of good nucleophiles an 8-I-2 species gives photoreaction via an S(N)1 mechanism. This is corroborated by the solvent dependence of the UV spectra, and the product composition upon photoreaction with bromide in varying concentration. Photoreaction of the iodonium salt in a chlorinated alkane yields (E)- and (Z)-beta-fluorostyrene in a Schiemann-type reaction. Reaction in toluene yields Friedel-Crafts products. The results of the photochemical reactions are compared to those of the thermal ones, and the implications of the differences are discussed. PMID- 11856010 TI - Diethyl chlorophosphite: a versatile reagent. AB - Diethyl chlorophosphite (DECP) was previously described as a reducing agent for nitro compounds to the corresponding amines (Fischer, B.; Sheihet, L. J. Org. Chem. 1998, 63, 393). Here, the utility of this reagent was extended to chemical conversions of other oxygenated functional groups. In this paper we report on the scope of the reaction of DECP with N-oxides, epoxides, sulfones, sulfoxides, hydroxylamines, ketoximes, and aldoximes. The chemoselectivity of DECP is described, and conditions for a stepwise multiple conversion of functional groups on the same molecule with this reagent are provided. PMID- 11856009 TI - Deactivation behavior and excited-state properties of (coumarin-4-yl)methyl derivatives. 2. Photocleavage of selected (coumarin-4-yl)methyl-caged adenosine cyclic 3',5'-monophosphates with fluorescence enhancement. AB - A series of axial and equatorial diastereomers of (coumarin-4-yl)methyl-caged adenosine cyclic 3',5'-monophosphates (cAMPs), 1-6, having methoxy, dialkylamino, or no substituent in the 6- and/or 7-positions, and their corresponding 4 (hydroxymethyl)coumarin photoproducts 7-12 have been synthesized. The photochemical and UV/vis spectroscopical properties (absorption and fluorescence) of 1-6 and 7-12 have been examined in methanol/aqueous HEPES buffer solution. Donor substitution in the 6-position causes a strong bathochromic shift of the long-wavelength absorption band, whereas substitution in the 7-position leads only to a weak red shift. The photochemical cleavage of the caged cAMPs was investigated, and the photoproducts were analyzed. Photochemical quantum yields, fluorescence quantum yields, and lifetimes of the excited singlet states were determined. The highest values of photochemical quantum yields (photo-S(N)1 mechanism) were obtained with caged cAMPs having a donor substituent in the 7 position of the coumarin moiety, caused by electronic stabilization of the intermediately formed coumarinylmethyl cation. With donor substitution in the 6 position, the resulting moderate electronic stabilization of the coumarinylmethyl cation is overcompensated by the strong bathochromic shift, reducing the energy gap between the excited-state S(1) and the corresponding coumarinylmethyl cation. The rate constant for the ester cleavage and liberation of cAMP is about 10(9) s( 1), estimated for the axial isomer of 6 by analysis of the fluorescence increase of the alcohol 12 formed upon laser pulse photolysis. PMID- 11856011 TI - Heterodimerization of dye-modified cyclodextrins with native cyclodextrins. AB - The heterodimerization behavior of dye-modified beta-cyclodextrins (1-6) with native cyclodextrins (CDs) was investigated by means of absorption and induced circular dichroism spectroscopy in an aqueous solution. Three types of azo dye modified beta-CDs (1-3) show different association behaviors, depending on the positional difference and the electronic character of substituent connected to the CD unit in the dye moiety. p-Methyl red-modified beta-CD (1), which has a 4 (dimethylamino)azobenzene moiety connected to the CD unit at the 4' position by an amido linkage, forms an intramolecular self-complex, inserting the dye moiety in its beta-CD cavity. It also associates with the native alpha-CD by inserting the moiety of 1 into the alpha-CD cavity. The association constants for such heterodimerization are 198 M(-1) at pH 1.00 and 305 M(-1) at pH 6.59, which are larger than the association constant of 1 for beta-CD (43 M(-1) at pH 1.00). Methyl red-modified 2, which has the same dye moiety as that for 1 although its substituent position is different from that of 1, does not associate even with alpha-CD due to the stable self-intramolecular complex, in which the dye moiety is deeply included in its own cavity of beta-CD. Alizarin yellow-modified CD (3), which has an azo dye moiety different from that of 1 and 2, caused a slight spectral variation upon addition of alpha-CD, suggesting that the interaction between 3 and alpha-CD is weak. On the other hand, phenolphthalein-modified beta CD (4), which forms an intermolecular association complex in its higher concentrations, binds with beta-CD with an association constant of 787 M(-1) at pH 10.80, where 4 exists as the dianion monomer in the absence of beta-CD. p Nitorophenol-modified beta-CDs (5 and 6), each having p-nitorophenol moieties with a different connecting part with an amido and amidophenyl group, respectively, associated with alpha-CD with association constants of 66 and 16 M( 1) for 5 and 6, respectively. The phenyl unit in the connecting part of 6 may prevent the smooth binding with alpha-CD. All these results suggest that the dye modified CDs, in which the dye part is not tightly included in its CD cavity, associate with the native CD to form heterodimer composed of two different CD units by inserting the dye moiety into the native CD unit. The resulting heterodimers have a cavity that can bind another appending moiety of host molecules. On this basis, more ordered molecular arrays or the supramolecular hereropolymers can be constructed. PMID- 11856012 TI - Synthesis of new beta-substituted meso-tetraphenylporphyrins via 1,3-dipolar cycloaddition reactions. 1. AB - The azomethine ylide generated from the reaction of (beta-formyl-meso tetraphenylporphyrinato)nickel(II) with N-methylglycine reacts with a range of dipolarophiles, yielding new beta-substituted-meso-tetraphenylporphyrins. The regio- and stereochemistry of the new compounds was established using one- and two-dimensional NMR techniques. PMID- 11856013 TI - Total synthesis of bafilomycin V(1): a methanolysis product of the macrolide bafilomycin C(2). AB - A synthesis of bafilomycin V(1), a methanolysis product of the macrolide natural product bafilomycin C(2), is described. The route utilizes chiral nonracemic allenylzinc reagents, prepared in situ from propargylic mesylates, to access key segments of this methyl ester. The acetylenic moieties of the derived homopropargylic alcohol adducts play an important role in further elaboration of these subunits. Final assemblage of the 25-carbon chain, containing 12 stereocenters, an alpha-methoxy Z,E 1,3-dienic ester, and an additional E,E 1,3 diene, was achieved through Stille coupling of an acetylene-derived vinyl stannane and vinyl iodide of approximate equal complexity. Attempted cyclization of several C15 hydroxy acid derivatives to the 16-membered lactone bafilomycin A(1), a potent inhibitor of vacuolar ATPases, could not be achieved. PMID- 11856015 TI - Transient resonance Raman and density functional theory investigation of the chlorine atom/carbon disulfide molecular complex involved in selective alkane photochlorination reactions. AB - The structure and bonding of the chlorine atom/carbon disulfide (CS(2)/Cl) complex involved in selective photochlorination reactions with alkanes was directly probed using transient resonance Raman spectroscopy. The experimental Raman vibrational frequencies were compared to those computed from density functional theory calculations for probable structures of the CS(2)/Cl complex. Our results indicate that the S [double bond] C [double bond] S...Cl complex species is responsible for the approximately 370 nm transient absorption band observed after ultraviolet photolysis of CCl(4) in the presence of CS(2). We discuss the structure and properties of the S [double bond] C [double bond] S...Cl complex and compare them with those for the related benzene/Cl and pyridine/Cl complexes. PMID- 11856014 TI - Synthesis of an optically active, bicyclic 2-pyridone dipeptide mimetic. AB - The eleven-step preparation of the bicyclic 2-pyridone dipeptide mimetic 1 [(3S) 6-(benzyloxycarbonylamino)-5-oxo-1,2,3,5-tetrahydroindolizine-3-carboxylic acid] in optically active form (60% ee) is described. Key steps in the synthesis of 1 include the osmium-catalyzed asymmetric dihydroxylation of olefin 13 [(6-but-3 enyl-2-methoxypyridin-3-yl)carbamic acid benzyl ester] and the intramolecular cyclization of protected diol 19 [(3'R)-[6-[4'-(tert-butyldimethylsilanyloxy)-3' hydroxybutyl]-2-methoxypyridin-3-yl]carbamic acid benzyl ester] to afford the pyridinium salt 20 [(3S)-[3-(tert-butyldimethylsilanyloxymethyl)-5-methoxy-2,3 dihydro-1H-indolizin-6-yl]carbamic acid benzyl ester trifuoromethanesulfonic acid salt]. Several alternate methods to prepare olefin 13 are also discussed. PMID- 11856016 TI - Preparation and structural study of the enantiomers of alpha,alpha' bis(trifluoromethyl)-9,10-anthracenedimethanol and its perdeuterated isotopomer, highly effective chiral solvating agents. AB - Enantiopure forms of alpha,alpha'-bis(trifluoromethyl)-9,10-anthracenedimethanol and the corresponding perdeuterated isotopomers were prepared. The conformational study was carried out by (1)H NMR, and the absolute configuration was determined by the X-ray study of the crystallized diastereoisomeric carbamate derivative. This compound was tested as a chiral solvating agent (CSA). The results showed very good discrimination for several racemic mixtures that improved other classical methods. The study of diastereomeric complexes was carried out by determination of the stoichiometry of the complex and the binding constant of the equilibrium. PMID- 11856017 TI - Metalation of 2-chloromethyl-2-oxazolines: synthesis of 1,2,3 tris(oxazolinyl)cyclopropanes and derivatives. AB - 2-Chloromethyl-2-oxazoline converts cleanly into trans-1,2,3 tris(oxazolinyl)cyclopropane upon treatment with strong bases such as LDA or KN(SiMe(3))(2). Deprotonation of the above cyclopropane followed by the addition of electrophiles allows the preparation of more functionalized tris(oxazolinyl)cyclopropanes. PMID- 11856018 TI - Asymmetric total synthesis of (+)-brefeldin A from (S)-lactate by triple chirality transfer process and nitrile oxide cycloaddition. AB - A novel synthesis of (+)-brefeldin A (1) has been accomplished on the basis of triple chirality transfer methodology, intramolecular ester enolate alkylation, and both intra- and intermolecular nitrile oxide cycloaddition strategies. PMID- 11856019 TI - Role of conformational effects on the regioselectivity of macrocyclic INOC reactions: two new asymmetric total syntheses of (+)-brefeldin A. AB - We have gained some insight into the role of conformational effects on the regioselectivity of the macrocyclic intramolecular nitrile oxide cycloaddition observed in our (+)-brefeldin A synthesis. During the course of this regiochemical study, we have developed two novel stereoselective and regioselective schemes for total synthesis of (+)-brefeldin A (i.e. intramolecular nitrile oxide cycloaddition-isomerization and intermolecular nitrile oxide cycloaddition-ring closing metathesis strategies). PMID- 11856020 TI - Stereocontrolled IMDA reaction of styrene derivatives. A way to enantiopure 3a,4,9,9a-tetrahydrobenz[f]isoindolines. AB - IMDA reactions on chiral perhydro-1,3-benzoxazines, derived from (-)-8-amino menthol, bearing a styrene substituent at C-2 acting as diene and an acryl amide acting as dienophile occur with high stereoselection and excellent chemical yields. After elimination of the chiral appendage, enantiopure 3a,4,9,9a tetrahydrobenz[f]isoindolines are prepared in this way. The effect of the substituents at both diene and dienophile are studied, showing that a methyl group at C-1 in the diene inhibited the reaction, while the ene adduct, instead of the IMDA product, was obtained when a methyl group was at C-2. PMID- 11856021 TI - Protonation studies of modified adenine and adenine nucleotides by theoretical calculations and (15)N NMR. AB - The acid/base character of nucleobases affects phenomena such as self association, interaction with metal ions, molecular recognition by proteins, and nucleic acid base-pairing. Therefore, the investigation of proton-transfer equilibria of natural and synthetic nucleos(t)ides is of great importance to obtain a deeper understanding of these phenomena. For this purpose, a set of ATP prototypes was investigated using (15)N NMR spectroscopy, and the corresponding adenine bases were investigated by theoretical calculations. (15)N NMR measurements provided not only acidity constants but also information on the protonation site(s) on the adenine ring and regarding the ratio of the singly protonated species in equilibrium. Substituents of different nature and position on the adenine ring did not change the preferred protonation site, which remained N1. However, for 2-thioether-ATP derivatives a mixed population of N1 and N7 singly protonated species was observed. Reduction of basicity of 0.4-1 pK(a) units relative to ATP was also observed for all evaluated ATP derivatives, except for 2-Cl-ATP, for which K(a) was ca. 10,000-fold lower. To explain the substitution-dependent variations in the experimental pK(a) values of the ATP analogues, gas-phase proton affinities (PA), Delta Delta G(hyd), and pK(a) values of the corresponding adenine bases were calculated using quantum mechanical methods. The computed PA and Delta Delta G(hyd) values successfully explained the experimental pK(a) values. A computational procedure for the prediction of accurate pK(a) values was developed using density functional theory and polarizable continuum model calculations. In this procedure, we developed a set of parameters for the polarizable continuum model that was fitted to reproduce experimental pK(a) values of nitrogen heterocycles. This method is proposed for the prediction of pK(a) values and protonation site(s) of purine analogues that have not been synthesized or analyzed. PMID- 11856022 TI - New insights into the acid-promoted reaction of caffeic acid and its esters with nitrite: decarboxylation drives chain nitrosation pathways toward novel oxime derivatives and oxidation/fragmentation products thereof. AB - In 0.05 M acetate buffer, pH 4, containing 1% methanol, caffeic acid (1a) (2 x 10(-3) M) reacted smoothly with nitrite (NO(2)(-)) (4 x 10(-3) M) to afford as main products the novel 2-hydroxy- and 2-methoxyaldoximes 7a,b, the 2-oxoaldoxime 9a, 3,4-dihydroxybenzoic acid, 3,4-dihydroxybenzaldehyde, and the known furoxan 3c and benzoxazinone 4b in smaller amounts. At lower 1a concentration (e.g., 1 x 10(-4) M), 7a was the main product, whereas with 0.1 M 1a and 0.5 M NO(2)(-) 3c and 9a were prevailing. At pH 2, 7a was still the most abundant product, together with 3,4-dihydroxybenzaldehyde and some 9a, whereas at pH 1 9a and 3,4 dihydroxybenzaldehyde were formed in higher yields. No evidence for ring nitration products, including the previously reported 4,5-dihydroxy-2 nitrobenzaldehyde, was obtained. At 2 x 10(-3) M concentration and at pH 4, caffeic acid methyl ester (1b) reacted with NO(2)(-) chiefly via ring nitration and/or dimerization to give 5a, the novel nitrated neolignan derivative 10, and the parent 6. Chlorogenic acid (1c) afforded only the ring nitrated derivative 5b. A unifying mechanism for the reaction of 1a and its esters with NO(2)(-) is proposed involving reversible formation of nitroso intermediates via chain nitrosation at the 2-position of the (E)-3-(3,4-dihydroxyphenyl)propenoic system. In the case of 1a, decarboxylation would drive the nitroso intermediates toward the formation of oximes 7a,b and 3c, reflecting nucleophilic addition of water, methanol, and NO(2)(-), and their oxidation or breakdown products, viz. 9a, 3,4 dihydroxybenzaldehyde, 3,4-dihydroxybenzoic acid, and the benzoxazinone 4b. In the case of esters 1b,c, to which decarboxylation is precluded, ring nitration or dimerization become the favored routes, triggered by preliminary oxidation at the catechol moiety. PMID- 11856023 TI - The mechanism of base-promoted HF elimination from 4-fluoro-4-(4 nitrophenyl)butan-2-one is E1cB. evidence from double isotopic fractionation experiments. AB - Leaving-group fluorine and secondary deuterium multiple kinetic isotope effects (KIEs) have been determined for the base-promoted HF elimination from the 4 fluoro-4-(4'-nitrophenyl)-(1,1,1,3,3-(2)H(5))butan-2-one. The fluorine KIE was determined by using the accelerator-produced short-lived radionuclide (18)F in combination with the naturally abundant (19)F. The (19)F substrate was labeled with (14)C in a remote position to enable radioactivity measurements of both substrates. The size of the determined fluorine KIE is 1.0009 +/- 0.0010 when acetate is used as base. The secondary deuterium KIEs are 1.009 +/- 0.017, 1.000 +/- 0.018, and 1.010 +/- 0.023 for formate, acetate, and imidazole, respectively. The magnitudes of these KIEs are significantly smaller compared to the corresponding KIEs that we recently reported for the protic substrate. This new data clearly demonstrates that the elimination proceeds via an E1cB mechanism. PMID- 11856024 TI - Structure and chemistry of apicidins, a class of novel cyclic tetrapeptides without a terminal alpha-keto epoxide as inhibitors of histone deacetylase with potent antiprotozoal activities. AB - Apicidins are a class of cyclic tetrapeptides that do not contain the classical electrophilic alpha-keto epoxide yet are potent (nM) inhibitors of histone deacetylase and antiprotozoal agents. These compounds showed broad-spectrum activities against the apicomplexan family of protozoa including Plasmodium sp (malarial parasite), Toxoplasma gondii, Cryptosporidium sp., and Eimeria sp. These cyclic peptides contain a beta-turn amino acid (R)-Pip or (R)-Pro, (S)-N methoxy Trp, (S)-Ile, or (S)-Val, and either (S)-2-amino-8-oxodecanoic acid or a modified (S)-2-amino-8-oxodecanoic acid. The isolation and structure elucidation of new apicidins from two Fusarium species, temperature-dependent NMR studies of apicidin, NMR and molecular modeling based conformation of the 12-membered macrocyclic ring, and selected chemical modifications of apicidin have been detailed in this paper. The cyclic nature of the peptide, the C-8 keto group, and the tryptophan are all critical for the biological activity. PMID- 11856025 TI - Methyl rotational barriers in amides and thioamides. AB - The methyl rotational barriers for a series of N-methyl-substituted amides and thioamides have been calculated at the MP2/6-311+G** level. A comparison of the N methylformamide and methyl formate barriers indicates that the H [bond] C(Me) [bond] N [bond] H eclipsed torsional arrangement destabilizes an amide by about 0.8 kcal/mol. A comparison of thioamides and amides showed the importance of steric repulsion between the sulfur and a methyl hydrogen in the Z-forms of the thioamides. The C [bond] N bond rotation transition states of the N,N-dimethyl amides have much larger methyl rotational barriers than found in the ground states. They can be attributed to the smaller CH(3)(-)N [bond] CH(3) bond angles in the transition states. PMID- 11856026 TI - Anionic cyclization approach toward perhydrobenzofuranone: stereocontrolled synthesis of the hexahydrobenzofuran subunit of avermectin. AB - A facile anionic cyclization approach toward stereocontrolled synthesis of the hexahydrobenzofuran subunit 3 of avermectin is described. As a model study, treatment of iodo compound 7 with n-BuLi at -100 degrees C effected metal-halogen exchange and subsequent anionic cyclization to afford perhydrobenzofuranone 8. For the total synthesis of subunit 3, compound 9 was dihydroxylated to give diol 10. Protection of the hydroxyl groups of diol 10 gave compound 11. Ketone 11 was then converted into the required enone 12 using Saegusa's protocol. On iodination followed by Luche reduction, enone 12 yielded alpha-iodo allylic alcohol 14, which on alkylation afforded ether 15. Conversion of the ester unit of 15 into a Weinreb amide group followed by anionic cyclization gave enone 17. 1,4-Addition of (MeOCH(2))(2)CuCNLi(2) to enone 17 followed by cleavage of the acetal unit afforded ketone 19. Preferential acetylation of the secondary alcoholic function of 19 afforded compound 20. The stereochemistry of 20 is confirmed by single crystal X-ray analysis. Elimination of HOAc from 20 gave the crucial olefin 21. Hydrolysis of the acetate unit of 21 followed by protection of the resulting alcoholic function yielded tert-butyldimethylsilyl ether 23. Introduction of a hydroxyl group at the ring junction of 23, using Davis's procedure, finally afforded the hexahydrobenzofuran subunit 3. PMID- 11856027 TI - Mechanistic investigation of a novel vitamin B(12)-catalyzed carbon [bond] carbon bond forming reaction, the reductive dimerization of arylalkenes. AB - In the presence of catalytic vitamin B(12) and a reducing agent such as Ti(III)citrate or Zn, arylalkenes are dimerized with unusual regioselectivity forming a carbon [bond] carbon bond between the benzylic carbons of each coupling partner. Dimerization products were obtained in good to excellent yields for mono and 1,1-disubstituted alkenes. Dienes containing one aryl alkene underwent intramolecular cyclization in good yields. However, 1,2-disubstituted and trisubstituted alkenes were unreactive. Mechanistic investigations using radical traps suggest the involvement of benzylic radicals, and the lack of diastereoselectivity in the product distribution is consistent with dimerization of two such reactive intermediates. A strong reducing agent is required for the reaction and fulfills two roles. It returns the Co(II) form of the catalyst generated after the reaction to the active Co(I) state, and by removing Co(II) it also prevents the nonproductive recombination of alkyl radicals with cob(II)alamin. The mechanism of the formation of benzylic radicals from arylalkenes and cob(I)alamin poses an interesting problem. The results with a one electron transfer probe indicate that radical generation is not likely to involve an electron transfer. Several alternative mechanisms are discussed. PMID- 11856028 TI - Alpha-(N-carbamoyl)alkylcuprate chemistry in the synthesis of nitrogen heterocycles. AB - The conjugate adducts obtained via coupling of alpha-(N-carbamoyl)alkylcuprates with alpha,beta-ynoates, alpha-allenyl esters, or alpha.beta-enoates or enimides undergo N-Boc deprotection and cyclization onto the ester functionality upon treatment with PhOH/TMSCl, catecholboron bromide, or trimethylsilyl triflate. This two-pot sequence provides synthetic routes to 4-alkylidinepyrrolidine-2 ones, 4-alkylidinepyrrolizidin-2-ones, and 4-alkylidineindolizidin-2-ones via allenyl esters; pyrrolin-2-ones, tetrahydropyrrolizin-2-ones, and tetrahydroindolizin-2-ones via alpha/beta-ynoates; pyrrolidin-2-ones, pyrrolizidin-2-ones, and indolizidin-2-ones via alpha,beta-enoates or alpha.beta enimides. The reluctance of gamma-carbamoyl-alpha,beta-enoates to undergo E/Z isomerization requires the use of (Z)-beta-iodo-alpha,beta-enoates readily prepared by the addition of HI to the alkynyl esters for the efficient preparation of pyrrolinones, tetrahydropyrrolizinones, and tetrahydroindolizinones. Utilization of omega-functionalized alpha,eta-ynoates or beta-iodo-alpha,beta-enoates allows for cyclization onto the omega-functionality providing for a synthetic route to quinolizidines. PMID- 11856029 TI - Tandem oligonucleotide synthesis on solid-phase supports for the production of multiple oligonucleotides. AB - More than one oligonucleotide can be synthesized at a time by linking multiple oligonucleotides end-to-end in a tandem manner on the surface of a solid-phase support. The 5'-terminal hydroxyl position of one oligonucleotide serves as the starting point for the next oligonucleotide synthesis. The two oligonucleotides are linked via a cleavable 3'-O-hydroquinone-O,O'-diacetic acid linker arm (Q linker). The Q-linker is rapidly and efficiently coupled to the 5'-OH position of immobilized oligonucleotides using HATU, HBTU, or HCTU in the presence of 1 equiv of DMAP. This protocol avoids introduction of phosphate linkages on either the 3' or 5'-end of oligonucleotides. A single NH(4)OH cleavage step can simultaneously release the products from the surface of the support and each other to produce free 5'- and 3'-hydroxyl termini. Selective cleavage of one oligonucleotide out of two sequences has also been accomplished via a combination of succinyl and Q linker linker arms. Tandem synthesis of multiple oligonucleotides is useful for producing sets of primers for PCR, DNA sequencing, and other diagnostic applications as well as double-stranded oligonucleotides. Tandem synthesis of the same sequence multiple times increases the yield of material from any single synthesis column for maximum economy in large-scale synthesis. This method can also be combined with reusable solid-phase supports to further reduce the cost of oligonucleotide production. PMID- 11856030 TI - Asymmetric synthesis of trans-2,3-piperidinedicarboxylic acid and trans-3,4 piperidinedicarboxylic acid derivatives. AB - Asymmetric syntheses of (2S,3S)-3-(tert-butoxycarbonyl)-2-piperidinecarboxylic acid (1b), (3R,4S)-4-(tert-butoxycarbonyl)-3-piperidinecarboxylic acid (2b), and their corresponding N-Boc and N-Cbz protected analogues 8a,b and 17a,b are described. Enantiomerically pure 1b has been synthesized in five steps starting from L-aspartic acid beta-tert-butyl ester. Tribenzylation of the starting material followed by alkylation with allyl iodide using KHMDS produces the key intermediate 5a in a 6:1 diastereomeric excess. Upon hydroboration, the alcohol 6a is oxidized, and the resulting aldehyde 7 is subjected to a ring closure via reductive amination, providing 1b in an overall yield of 38%. Optically pure 2b has been synthesized beginning with N-Cbz-beta-alanine. The synthesis involves the induction of the first stereogenic center using Evans's chemistry and sequential LDA-promoted alkylations with tert-butyl bromoacetate and allyl iodide. Further elaboration by ozonolysis and reductive amination affords 2b in an overall yield of 28%. PMID- 11856031 TI - Biosynthesis of zeaxanthin via mevalonate in Paracoccus species strain PTA-3335. A product-based retrobiosynthetic study. AB - Cultures of the zeaxanthin-producing bacterium Paracoccus species strain PTA-3335 (formerly Flavobacterium) were grown with supplements of (13)C-labeled glucose. Zeaxanthin was isolated and analyzed by (13)C NMR spectroscopy. The data showed that the isoprenoid precursors of zeaxanthin were biosynthesized via the mevalonate pathway. The microorganism was found to utilize glucose mainly via the Entner-Doudoroff pathway. PMID- 11856032 TI - Synthesis of spiro-fused (C5)-isoxazolino-(C4)-pyrazolones (1-oxa-2,7,8 triazaspiro[4,4]-2,8-dien-6-ones) via 1,3-dipolar cycloaddition and cycloelimination. AB - An efficient and selective method for the synthesis of spiro-fused (C5) isoxazolino-(C4)-pyrazolones (C) is reported. The process consists of utilizing the Baylis-Hillman reaction-or a quicker, stepwise MAC procedure-to give I followed by 1,3-dipolar cycloaddition and Swern oxidation to give beta-ketoesters H, which were condensed with hydrazine derivatives to provide hydrazones that underwent cycloelimination. These novel spiro-fused (C5)-isoxazolino-(C4) pyrazolones were confirmed by spectroscopic analysis as well as single-crystal X ray of 5. We also concluded that all condensations/cycloeliminations, except with hydrazine itself, were more effective with catalysts or higher reaction temperatures. For example, TiCl(4) was an efficient catalyst for hydrazone formation and cycloelimination with methylhydrazine, while phenyl-, benzyl-, and (4-methoxyphenyl)hydrazine reacted effectively without catalyst in refluxing xylene. PMID- 11856033 TI - Acylation of 2,5-dimethoxycarbonyl[60]fulleropyrrolidine and synthesis of its multifullerene derivatives. AB - 2,5-Dimethoxycarbonyl[60]fulleropyrrolidine (1) is acylated with various chlorocarbonyl compounds to give fullerene derivatives with the general formula C(60)(MeOOCCH)(2)NC(O)R, R = (CH(2))(5)Br, (CH(2))(8)C(O)Cl (3), (CH(2))(4)C(O)Cl, or cis-C(6)H(4)(C(O)Cl. The monoacylated sebacoyl derivative 3 readily reacts with alcohols and amines such as methanol, diethylamine, glycine methyl ester, and aza-18-crown-6 through the remaining chlorocarbonyl group. Chromatography of 3 on silica gel converts it into the corresponding acid C(60)(MeOOCCH)(2)NC(O)(CH(2))(8)COOH (4). Treating 4 with PCl(5) regenerates the precursor 3 quantitatively. Piperazine reacts with 4 in the presence of DCC and BtOH to form a bisfullerene derivative in which two sebacoyl chains and the piperazine act as the bridge between two molecules of 1. Other molecules with multifunctional groups react with 4 similarly to form multifullerene derivatives. NMR data indicate that the rotation of the relatively bulky phthaloyl group is hindered around the amide bond N [bond] C(O), the rotation barrier of which is 15.06 kcal/mol. The relative stereochemistry of the 2,5-dimethoxycarbonyl groups is established by (1)H NMR spectra and further confirmed by resolution of the enantiomeric 2,5-trans-isomer of the starting material 1. PMID- 11856034 TI - Arabinofuranosides from mycobacteria: synthesis of a highly branched hexasaccharide and related fragments containing beta-arabinofuranosyl residues. AB - The synthesis of 11 oligosaccharides (4-14) containing beta-arabinofuranosyl residues is reported. The glycans are all fragments of two polysaccharides, arabinogalactan and lipoarabinomannan, which are found in the cell wall complex of mycobacteria. In the preparation of the targets, the key step was a low temperature glycosylation reaction that installed the beta-arabinofuranosyl residues with good to excellent stereocontrol. PMID- 11856035 TI - Synthesis of 3,5- and 3,6-linked calix[n]naphthalenes. AB - The preparation of calix[n]naphthalenes from derivatives of 2,7 dihydroxynaphthalene is described. 1,8-Dialkyl substitution is used to direct the regiochemistry of the acid-catalyzed condensation reactions. Acyclic peri substituents lead to a 3,5-linked calix[3]naphthalene, whereas cyclic peri substituents give predominantly a calix[5]naphthalene with the corresponding 3,6 linkage. The 3,6-linked calix[4]naphthalene is prepared in pure form by a dimerization strategy. PMID- 11856036 TI - Expedient solid-phase synthesis of fluorogenic protease substrates using the 7 amino-4-carbamoylmethylcoumarin (ACC) fluorophore. AB - A highly efficient solid-phase synthesis method for the preparation of fluorogenic protease substrates based upon the bifunctional leaving group 7-amino 4-carbamoylmethylcoumarin (ACC) is reported. Methods for the large-scale preparation of the novel fluorogenic leaving-group ACC are provided (Scheme 1). Detailed procedures are also provided for loading a diverse set of amino acids to support-bound ACC in good yields and with minimal racemization. Finally, procedures are included for the preparative synthesis of optimized ACC substrates for HIV-1 protease and plasmin. PMID- 11856037 TI - Anthracene derivatives and the corresponding dimers with TEMPO radicals. AB - Anthracene derivatives with several TEMPO radicals (2-4, 10) were prepared, and each photodimerization reaction was investigated. Although the photodimerization was unsuccessful in obtaining the dimers of anthracenes 2 and 3, which could be alternatively prepared in a stepwise manner, the photodimers of anthracenes 4 and 10 were available by the direct photoreaction. The dissociation reaction of the dimers proceeded well by heating them in solution to give the corresponding monomers in each case, and thus the reversible system could be constructed in the latter two systems. While no large difference was observed in their magnetic behaviors between the monomer/dimer pair of 4 and 8, an intriguing difference was found in the magnetic behaviors for the pair of 10 and 11 from ferromagnetic interactions in 10 to the variable magnetic interactions in 11 depending on the solvent molecules incorporated in the crystals. PMID- 11856038 TI - Radical [3 + 2] cycloaddition reaction with various alkenes using iodomethylcyclopropane derivatives as new homoallyl radical precursors. AB - Radical iodine atom transfer [3 + 2] cycloaddition with various alkenes using dimethyl 2-(iodomethyl)cyclopropane-1,1-dicarboxylate and 1,1-bis(phenylsulfonyl) 2-(iodomethyl)cyclopropane as new precursors of a homoallyl radical species smoothly proceeds to give functionalized cyclopentane derivatives in good yields. PMID- 11856039 TI - Bond dissociation enthalpies of polyphenols: the importance of cooperative effects. AB - The hydrogen-oxygen bond dissociation energies of 3,5-di-tert-butylcatechol, 2,5 di-tert-pentylhydroquinone, propyl gallate, and octyl gallate, which represent model compounds of three important classes of naturally occurring antioxidants, have been measured by an EPR equilibration technique, and the factors determining their values have been clarified. The excellent antioxidant activity of the these polyphenols is largely due to the stabilization of the aroxyl radical due to the formation of an intramolecular hydrogen bond. PMID- 11856040 TI - Highly chemoselective hydrogenolysis of iodoarenes. AB - The catalytic hydrodehalogenation reaction using molecular hydrogen and Pd/C has been revisited. It is shown that the speed of removal of halogen increases with increasing electronegativity I < Br < Cl. Nevertheless, selective dehydrohalogenation in compounds containing other reducible functions can be achieved only with iodine and not with bromine or chlorine. Selective deiodination of iodobenzophenone could be accomplished without reducing the carbonyl group. Hydrogenolysis of azidoiodoaromatic compounds to the corresponding azido compounds is high yielding. This selectivity was exploited for the labeling of benzophenone- and azido-containing compounds by deuterium and tritium. PMID- 11856041 TI - Multicomponent reactions involving 2-methyleneaziridines: rapid synthesis of 1,3 disubstituted propanones. AB - Ring opening of 1-alkyl-2-methyleneaziridines 1 or 2 is accomplished with organocopper reagents (R(2)CuLi or RMgX/CuI) in the presence of boron trifluoride diethyl etherate giving 1-substituted propan-2-ones 3-9 in 42-88% yield. Ring opening with RMgCl/CuI in the absence of the Lewis acid allows further alkylation of the metalloenamine (metalated imine) intermediate in a regiocontrolled manner. The sequential formation of two new intermolecular carbon-carbon bonds in this reaction provides a rapid entry into a variety of 1,3-disubstituted propan-2 ones, including 11 and 14-23. The scope and mechanism of this multicomponent reaction (MCR) has been assessed. It is established that this MCR tolerates alkyl, aryl, and benzylic Grignard reagents and a wide range of electrophiles, including alkyl iodides, bromides, and tosylates, as well as epoxides and aldehydes. In addition, gem-dimethyl substitution on the exocyclic double bond of the 2-methyleneaziridine is tolerated. This MCR has been applied to the one-pot synthesis of (Z)-6-heneicosen-11-one, 25, an important sex attractant of the Tussock moth. Using 3-deuterio-1-(1-phenylethyl)-2-methyleneaziridine, 26, we determined that this MCR occurs predominantly by direct ring opening at the sp(3) hybridized aziridine carbon atom (C-3). PMID- 11856042 TI - Synthesis and mode of action of (125)I- and (3)H-labeled thieno[2,3-c]pyridine antagonists of cell adhesion molecule expression. AB - A series of thieno[2,3-c]pyridine antagonists of cell adhesion molecule (CAM) expression, such as A-205804 (1) and A-249377 (2), selectively suppressed the induced expression of E-selectin and ICAM-1 over VCAM-1. In an effort to explore the biological mechanism of action of these inhibitors, we synthesized (125)I- and (3)H-labeled thieno[2,3-c]pyridines 5 and 6. An isolated diazonium tetrafluoroborate salt efficiently trapped Na(125)I on very small scale (7.5 microg of Na(125)I), providing the corresponding (125)I-labeled thieno[2,3 c]pyridine in modest yield. Preliminary mechanistic investigations using these radiolabeled compounds revealed that, upon incubation with human umbilical vein endothelial cells (HUVECs), these inhibitors of CAM expression translocated to the cell nucleus and were noncovalently associated with macromolecules of molecular weight greater than 650 kDa. PMID- 11856043 TI - Hydroxymethyl group conformation in saccharides: structural dependencies of (2)J(HH), (3)J(HH), and (1)J(CH) spin-spin coupling constants. AB - Experimental and theoretical methods have been used to correlate (2)J(HH) and (3)J(HH) values within the exocyclic hydroxymethyl groups (CH(2)OH) of saccharides with specific molecular parameters, and new equations are proposed to assist in the structural interpretation of these couplings. (3)J(HH) depends mainly on the C-C torsion angle (omega) as expected, and new Karplus equations derived from J-couplings computed from density functional theory (DFT) in a model aldopyranosyl ring are in excellent agreement with experimental values and with couplings predicted from a previously reported general Karplus equation. These results confirm the reliability of DFT-calculated (1)H-(1)H couplings in saccharides. (2)J(HH) values depend on both the C-C (omega) and C-O (theta) torsions. Knowledge of the former, which may be derived from other parameters (e.g., (3)J(HH)), allows theta to be evaluated indirectly from (2)J(HH). This latter approach complements more direct determinations of theta from (3)J(HCOH) and potentially extends these more conventional analyses to O-substituted systems lacking the hydroxyl proton. (1)J(CH) values within hydroxymethyl fragments were also examined and found to depend on r(CH), which is modulated by specific bond orientation and stereoelectronic factors. These latter factors could be largely, but not completely, accounted for by C-C and C-O torsional variables, leading to only semiquantitative treatments of these couplings (details discussed in the Supporting Information). New equations pertaining to (2)J(HH) and (3)J(HH) have been applied to the analysis of hydroxymethyl group J-couplings in several mono- and oligosaccharides, yielding information on C5-C6 and/or C6-O6 rotamer populations. PMID- 11856044 TI - Density functional theory study of the cycloaddition reaction of furan derivatives with masked o-benzoquinones. Does the furan act as a dienophile in the cycloaddition reaction? AB - The molecular mechanism for the cycloaddition reaction between 2-methylfuran and a masked o-benzoquinone has been characterized using quantum mechanical calculations at the B3LYP/6-31G theory level. An analysis of the results on the reaction pathway shows that the reaction takes place along a polar stepwise mechanism. The first and rate-determining step corresponds to the nucleophilic attack of the furan ring on the doubly conjugated position of the 2,4-dienone system present at the masked o-benzoquinone to give a zwitterionic intermediate. Closure of this intermediate affords the formally [2 + 4] cycloadduct. For the second step two reactive channels have been characterized corresponding to the formation of the formally [2 + 4] and [4 + 2] cycloadducts. Analysis of the energetic results indicates that while the first is the meta regiocontrolling and endo stereocontrolling step, the second one is responsible for the formation of the unexpected formally [2 + 4] cycloadduct. The global and local electrophilicity/nucleophilicity power of the reactants and intermediate have been evaluated to rationalize these results. Density functional theory analysis for these cycloadditions is in complete agreement with the experimental outcome, explaining the reactivity and selectivity of the formation of the formally [2 + 4] cycloadducts. PMID- 11856045 TI - Reduction of organic halides with lanthanum metal: a novel generation method of alkyl radicals. AB - Results of the reaction of alkyl halides with lanthanum metal have been shown. The reduction of alkyl iodide with 1/3 equiv of lanthanum metal efficiently proceeded to give the corresponding reductive dimerized products along with the formation of reduction and dehydroiodination products. In the case of alkyl bromides and chlorides, the reaction did not proceed under the same reaction conditions as that of alkyl iodides; however, the reaction was dramatically promoted by the addition of a catalytic amount of iodine. A reaction pathway including alkyl radicals was suggested. PMID- 11856046 TI - Catalytic radical addition of carbonyl compounds to alkenes by Mn(II)/Co(II)/O(2) system. AB - The radical addition of enolizable carbonyl compounds such as malonates and malononitrile to alkenes was successfully achieved through a catalytic process using the Mn(II)/Co(II)/O(2) system to afford the corresponding adducts in fair to good yields. Dimethyl malonate added to 1,5-cyclooctadiene to produce a fused bicycle compound. PMID- 11856047 TI - Palladium(II)-catalyzed highly regio- and diastereoselective cyclization of difunctional allylic N-tosylcarbamates. A convenient synthesis of optically active 4-vinyl-2-oxazolidinones and total synthesis of 1,4-dideoxy-1,4-imino-L xylitol. AB - A Pd(II)-catalyzed cyclization of difunctional allylic N-tosyl carbamates in the presence of halide ions was developed with high regio- and diastereoselectivity. The reaction involves aminopalladation of alkene and beta-heteroatom elimination to regenerate Pd(II) species. When the readily available homochiral alcohols were used as substrates, highly optically active 4-vinyl-2-oxazolidinones were easily obtained. The utility of this method was exemplified by the convenient synthesis of 1,4-dideoxy-1,4-imino-L-xylitol. PMID- 11856048 TI - Asymmetric 1,3-dipolar cycloaddition of N-metalated azomethine ylides to methyl (S)-2-(p-tolylsulfinyl)acrylate. Synthesis of optically pure 2,4,5-trisubstituted 2,5-dihydro-1H-pyrroles. AB - The first 1,3-dipolar reaction of azomethine ylides with optically pure vinyl sulfoxide are reported. The presence of the sulfinyl group increase the reactivity of the acrylate moiety as a dipolarophile, and the reactions evolve with complete regio- and endo-selectivities. Nevertheless, mixtures of the two diastereoisomers 4 and 5 (75-88% de) resulting from the anti dipole/s-cis dipolarophile and syn dipole/s-trans dipolarophile approaches, respectively, are obtained. The stereoselectivity can be controlled by using THF or MeCN as solvents or by changing the reaction temperature in MeCN. After separation of the cycloadducts, optically pure 2,5-dihydro-1H-pyrroles are easily obtained by pyrolytic desulfinylation. PMID- 11856050 TI - Reactivity of stable trifluoroacetaldehyde hemiaminals. 2. Generation and synthetic potentialities of fluorinated iminiums. AB - Under Lewis acid activation, hemiaminals of trifluoroacetaldehyde and related (fluoroalkyl)aldehydes generate iminium species that can react with various nucleophiles to provide fluorinated amines. PMID- 11856049 TI - Synthesis of homoceramides, novel ceramide analogues, and their lack of effect on the growth of hippocampal neurons. AB - The synthesis of a new series of D-erythro-homoceramide analogues is described. Several synthetic approaches were investigated. Homoceramides can be successfully synthesized from L-homoserine as chiral building block and a protected Weinreb amide as a key intermediate. The synthesis of short-chain analogues with a heptyl side chain, as well as with a phenyl residue in the sphingoid part (instead of the naturally occurring tridecyl side chain), was effected. The homoceramides 15 17 and 24 were investigated for their potential to reverse the inhibitory effect of fumonisin B(1) on axonal growth. Unfortunately, none of the tested compounds showed any biological activity due to their lack of metabolism to glucosylhomoceramide. PMID- 11856051 TI - An efficient synthesis of 4-(phenylsulfonyl)-4H-furo[3,4-b]indoles. AB - The fused heterocycle 4-(phenylsulfonyl)-4H-furo[3,4-b]indole, which is an indole 2,3-quinodimethane synthetic analogue, is prepared in five steps from indole in 46% yield. A similar sequence is used to synthesize C-3 derivatives (3-methyl, 3 phenyl, and 3-heptyl). Thus, indole-3-carbaldehyde (1) is protected as the N phenylsulfonyl derivative 2 and converted to the ethylene acetal 6. Lithiation at C-2 followed by treatment with an aldehyde affords the expected hydroxy acetals 7 and 8. Exposure to acid effects cyclization to the furoindoles 5 and 9. Furthermore, C-1 lithiation of furo[3,4-b]indole 9c followed by treatment with methyl iodide affords disubstituted furo[3,4-b]indole 10. PMID- 11856052 TI - New accelerated strategy for the synthesis of poly(ether ketone) dendrons. AB - A new AB(4)-type hypermonomer with four activated aromatic fluorines was prepared and converted to hyperbranched poly(ether ketones) by nucleophilic aromatic substitution of these fluorine atoms with phenolates. The preparation and application of the AB(4) hypermonomer for the accelerated synthesis of a family of poly(ether ketone) dendrons G2-G4 in good yield are described. PMID- 11856053 TI - Reaction of N,N-dimethylselenocarbamoyl chloride with nucleophiles. Preparation of diselenocarbamates, selenothiocarbamates, and selenoureas. AB - Reactions of N,N-dimethylselenocarbamoyl chloride prepared by using LiAlHSeH with nucleophiles such as lithium alkylselenolate, lithium alkylthiolate, and amines afford the corresponding diselenocarbamates, selenothiocarbamates, and selenoureas. The crystal structure of the Se-phenyl N,N-dimethyldiselenocarbamate was also determined by X-ray diffraction. PMID- 11856054 TI - Regiochemistry of the photostimulated reaction of the phthalimide anion with 1 iodoadamantane and tert-butylmercury chloride by the S(RN)1 mechanism. AB - The photostimulated reaction of the phthalimide anion (1) with 1-iodoadamantane (2) gave 3-(1-adamantyl) phthalimide (3) (12%) and 4-(1-adamantyl) phthalimide (4) (45%), together with the reduction product adamantane (AdH) (21%). The lack of reaction in the dark and inhibition of the photoinduced reaction by p dinitrobenzene, 1,4-cyclohexadiene, and di-tert-butylnitroxide indicated that 1 reacts with 2 by an S(RN)1 mechanism. Formation of products 3 and 4 occurs with distonic radical anions as intermediates. The photoinduced reaction of anion 1 with tert-butylmercury chloride (10) affords 4-tert-butylphthalimide (11) as a unique product. By competition experiments toward 1, 1-iodoadamantane was found to be ca. 10 times more reactive than tert-butylmercury chloride. PMID- 11856055 TI - Synthesis of novel L-2',3'-dideoxy-2'-trifluoromethyl-4'- thiocytidines from alpha-trifluoromethyl-alpha,beta-unsaturated ester. AB - A short and efficient synthesis of L-2',3'-dideoxy-2'-trifluoromethyl-4' thiocytidines is described. (2R,4S/2S,4S)-5-(tert-Butyldimethylsiloxy)-2 trifluoromethylpentan-4-olide (3a and 3b) are prepared from alpha-trifluoromethyl alpha,beta-unsaturated ester (1) in three steps and converted to compounds 6a and 6b. The corresponding 1-O-acetyl derivatives 8a and 8b were obtained via the usual Pummerer rearrangement from 6a and 6b in two steps, which were in turn used to synthesize L-4'-thiocytidines 10a and 10b. PMID- 11856056 TI - Amphidinolide W, a new 12-membered macrolide from dinoflagellate Amphidinium sp. AB - A new cytotoxic 12-membered macrolide, amphidinolide W (1), has been isolated from a marine dinoflagellate Amphidinium sp., and the structure was elucidated by spectroscopic data including (13)C-(13)C INADEQUATE correlations for its (13)C enriched sample. The absolute stereochemistry of 1 was assigned by combination of J-based configuration analysis and modified Mosher method. Amphidinolide W (1) is the first macrolide without an exomethylene unit among all amphidinolides obtained so far. PMID- 11856057 TI - Development of new chiral building blocks for synthesis of bicyclo[3.3.0]octane compounds. AB - Ti(II)-mediated tandem cyclization of (E)-5-(tert-butyldimethylsilyloxy)-8 trimethylsilyl-2-octen-7-ynoate (7) prepared from commercially available optically active epichlorohydrin (2) proceeded diastereoselectively to provide 7 (tert-butyldimethylsilyloxy)-2-trimethylsilylbicyclo[3.3.0]oct-1-en-3-one (1), which serves as a useful chiral building block or intermediate to prepare a variety of compounds having a bicyclo[3.3.0]octane framework. PMID- 11856058 TI - A simple and efficient chemoselective method for the catalytic deprotection of acetals and ketals using bismuth triflate. AB - Bismuth triflate is a highly efficient catalyst (0.1-1 mol %) for the deprotection of acetals and ketals. The procedure is very facile and selective for acetals derived from ketones and conjugated aldehydes. tert Butyldimethylsilyl ethers are stable to the reaction conditions. The highly catalytic nature of bismuth triflate and the use of a relatively nontoxic solvent system (THF/H(2)O) make this procedure particularly attractive for large-scale synthesis. PMID- 11856059 TI - Calix[4]azulene. AB - Azulene reacts with paraformaldehyde in the presence of florisil to give excellent yields of calix[4]azulene. PMID- 11856060 TI - Expedient, stereocontrolled synthesis of (+)-compactin lactone via intramolecular Reformatsky reaction. AB - A concise, stereodefined synthesis of compactin lactone is described. Samarium(II) iodide mediated intramolecular Reformatsky reaction is the key step in the synthesis. PMID- 11856061 TI - The first example of a singlet oxygen induced double bond migration during sulfide photooxidation. Experimental evidence for sulfone formation via a hydroperoxy sulfonium ylide. AB - The first example of the formation of a sulfone concomitant with double bond migration during photooxidation of a sulfide is reported. Evidence is presented which demonstrates that the double bond migration is not a result of a prior acid catalyzed rearrangement of an unrearranged sulfone precursor. This unusual observation is used to argue that the sulfone is formed via rearrangement of a hydroperoxy sulfonium ylide intermediate. PMID- 11856062 TI - Enantioselective synthesis of (2S,3R)-3-hydroxy-3-methylproline, a novel amino acid found in polyoxypeptins. AB - The enantioselective synthesis of (2S,3R)-3-hydroxy-3-methylproline (3) was achieved by the Sharpless AD, regioselective opening of cyclic sulfate by NaN(3) and intramolecular ring-closing reaction. The reported route has the advantage of a high overall yield and good enantiomeric purity, as well as starting from readily available chemical substrates and inexpensive reagents. PMID- 11856063 TI - Efficient and convenient nonaqueous workup procedure for the preparation of arylboronic esters. AB - An efficient one-pot synthetic protocol for the synthesis of arylboronic esters has been established. The concentrated addition mixture of trimethylborate with aryl Grignard reagents was treated with low molecular weight diols (ethylene glycol, 1,3-propandiol) and toluene, the corresponding arylboronic esters were isolated in a convenient way with high yields. The diols not only serve as water replacement for the workup step, but also as well as the reagent for the preparation of arylboronic esters. PMID- 11856064 TI - Age- and gender-related test performance in community-dwelling elderly people: Six-Minute Walk Test, Berg Balance Scale, Timed Up & Go Test, and gait speeds. AB - BACKGROUND AND PURPOSE: The interpretation of patient scores on clinical tests of physical mobility is limited by a lack of data describing the range of performance among people without disabilities. The purpose of this study was to provide data for 4 common clinical tests in a sample of community-dwelling older adults. SUBJECTS: Ninety-six community-dwelling elderly people (61-89 years of age) with independent functioning performed 4 clinical tests. METHODS: Data were collected on the Six-Minute Walk Test (6MW), Berg Balance Scale (BBS), and Timed Up & Go Test (TUG) and during comfortable- and fast-speed walking (CGS and FGS). Intraclass correlation coefficients (ICCs) were used to determine the test-retest reliability for the 6MW, TUG, CGS, and FGS measurements. Data were analyzed by gender and age (60-69, 70-79, and 80-89 years) cohorts, similar to previous studies. Means, standard deviations, and 95% confidence intervals for each measurement were calculated for each cohort. RESULTS: The 6MW, TUG, CGS, and FGS measurements showed high test-retest reliability (ICC [2,1]=.95-.97). Mean test scores showed a trend of age-related declines for the 6MW, BBS, TUG, CGS, and FGS for both male and female subjects. DISCUSSION AND CONCLUSION: Preliminary descriptive data suggest that physical therapists should use age-related data when interpreting patient data obtained for the 6MW, BBS, TUG, CGS and FGS. Further data on these clinical tests with larger sample sizes are needed to serve as a reference for patient comparisons. PMID- 11856065 TI - Is base of support greater in unsteady gait? AB - BACKGROUND AND PURPOSE: We investigated dynamic interfoot distance (IFD) throughout the gait cycle in people with unsteady gait caused by vestibulopathy and in people without known neuromuscular pathology. We expected that the subjects with unsteady gait would use a greater IFD than subjects without neuromuscular pathology and that this IFD would be correlated with other measures of locomotor stability. SUBJECTS AND METHODS: Simultaneous whole-body (11 segment) dynamic kinematic data were collected from 22 subjects with vestibulopathy and 22 subjects without known neuromuscular pathology who were matched for age, height, weight, and body mass index. Two trials each of the participants' gait at preferred speed and paced gait at 120 steps/min were analyzed with a repeated-measures design with multiple dependent variables. Quantitative data were analyzed descriptively and with inferential statistics. RESULTS: Interfoot distance at preferred gait speed did not differentiate unsteady subjects with vestibulopathy from the comparison subjects. Paced gait IFD total range and IFD in single-limb support differed between groups, but IFD at heel-strike did not. However, IFD at heel-strike, the traditional measure of "base-of-support width," was correlated with measurements of whole-body center-of gravity stability (r=.32-.55). DISCUSSION AND CONCLUSION: Gait at preferred speed permitted the unsteady subjects and the comparison subjects to select similar IFD values, but at the cost of slower gait in the unsteady subjects. When required to walk at a "normal" pace of 120 steps/min, subjects with vestibulopathy increased their IFD. These data suggest that wide-based gait alone cannot differentiate between subjects with and without balance impairments. Base of support and other whole-body kinematic variables are mechanical compensations of vestibulopathic instability. Further studies are needed to determine whether development of active control of these whole-body control variables can occur after vestibular rehabilitation. PMID- 11856066 TI - Kinematic analysis of kicking movements in preterm infants with very low birth weight and full-term infants. AB - BACKGROUND AND PURPOSE: Study of kicking development provides important information to understand how early spontaneous movements change in infants as they acquire voluntary control. Researchers have investigated the kicking movements of preterm infants; however, the movement patterns that they have described were inconsistent. The purpose of this study, therefore, was to examine the development of kicking movements with kinematic analysis in preterm infants with very low birth weight (VLBW) and full-term infants. SUBJECTS AND METHODS: Twenty-two infants with VLBW who were divided into low gestational age (gestational age of <30 weeks, n=9) and high gestational age (gestational age of >or=30 weeks, n=13) classes and 22 full-term infants were evaluated during kicking movements using 4 synchronized cameras and 3-dimensional kinematic analysis when the infants were 2 and 4 months of corrected age. RESULTS: The infants with VLBW and a high gestational age showed similar kicking movements compared with the full-term infants. In contrast, the infants with VLBW and a low gestational age exhibited a higher kick frequency and a shorter flexion phase at 4 months of corrected age. They also exhibited a higher hip-knee correlation and lower variability in the interlimb coordination pattern at 2 and 4 months of corrected age. DISCUSSION AND CONCLUSION: The findings indicate that infants with VLBW, particularly those with a low gestational age, have age-related differences in movement organization and coordination of kicking compared with full-term infants. PMID- 11856067 TI - Assessing the need for change in clinical education practices. AB - The purposes of this perspective article are to identify areas of need within clinical education, to describe various models and tools that are proposed and utilized in clinical education, and to explore the extent to which these models and tools might meet the identified needs of clinical education. A synthesis of the literature suggests that the clinical education process in physical therapy currently is characterized by 7 primary needs and that 10 models currently exist to guide the general process or to provide specific tools and practices to enhance its effectiveness. Roles and relationships are critical components in successful clinical education. Theory suggests that clinical educators and students should engage in an intentional, structured process of changing roles during the course of the clinical education experience and that nontechnical competencies such as communication, collaboration, and reflection are crucial for effective practice and may be developed in the clinical education setting. Developing a clearer understanding of the current status of physical therapy clinical education can assist clinical educators in the use of the available models and tools or in developing a new model that addresses potentially unique needs. PMID- 11856069 TI - Not "financing a sham". PMID- 11856070 TI - Don't disregard ultrasound yet--the jury is still out. PMID- 11856071 TI - Thera-Band Tubing. PMID- 11856072 TI - "Clinical education" versus clinical education. PMID- 11856068 TI - Pathophysiological tissue changes associated with repetitive movement: a review of the evidence. AB - Work-related musculoskeletal disorders (WMSDs) represent approximately one third of workers' compensation costs in US private industry, yet estimates of acceptable exposure levels for forceful and repetitive tasks are imprecise, in part, due to lack of measures of tissue injury in humans. In this review, the authors discuss the scope of upper-extremity WMSDs, the relationship between repetition rate and forcefulness of reaching tasks and WMSDs, cellular responses to injury in vivo and in vitro, and animal injury models of repetitive, forceful tasks. The authors describe a model using albino rats and present evidence related to tissue injury and inflammation due to a highly repetitive reaching task. A conceptual schematic for WMSD development and suggestions for further research are presented. Animal models can enhance our ability to predict risk and to manage WMSDs in humans because such models permit the direct observation of exposed tissues as well as motor behavior. PMID- 11856073 TI - Non-surgical treatment of hepatocellular carcinoma. AB - Primary hepatocellular cancer is a disease with a poor prognosis for which there is little consensus on treatment and a paucity of comparative trials. The coexistence of cancer with cirrhosis complicates treatment, and also confers a high risk for the development of further tumours. Surgery, either by hepatic resection or orthotopic liver transplantation, is only a feasible option in a minority of patients. This article surveys the non-surgical approaches to the treatment of hepatocellular cancers-local ablation techniques, arterial embolization with and without chemotherapy, conventional chemotherapy and hormonal modulation, and targeted and external irradiation. PMID- 11856074 TI - Transient lower oesophageal sphincter relaxations--a pharmacological target for gastro-oesophageal reflux disease? AB - The oesophago-gastric junction functions as an anti-reflux barrier preventing increased exposure of the oesophageal mucosa to gastric contents. Failure of this anti-reflux barrier results in gastro-oesophageal reflux disease, and may lead to complications such as oesophagitis, Barrett's oesophagus and eventually oesophageal carcinoma. Recent studies have suggested that transient lower oesophageal sphincter relaxation is the main mechanism underlying gastro oesophageal reflux. It involves a prolonged relaxation of the lower oesophageal sphincter, mediated by a vago-vagal neural pathway, synapsing in the brainstem. Several drugs, such as atropine, baclofen and loxiglumide, have been shown to reduce the rate of transient lower oesophageal sphincter relaxations and concomitantly the number of reflux episodes. These findings illustrate that transient lower oesophageal sphincter relaxations may represent a potential new target for the pharmacological treatment of gastro-oesophageal reflux disease. It is possible that the reduction in the number of transient lower oesophageal sphincter relaxations may also contribute to the beneficial effect of fundoplication and new endoscopic anti-reflux procedures. It should be emphasized, however, that other factors, such as low lower oesophageal sphincter pressure, the presence of a hiatal hernia and impaired oesophageal peristalsis, are also of great importance. Therefore, whether the targeting of transient lower oesophageal sphincter relaxations is the 'golden bullet' in anti-reflux therapy remains to be proven, as evidence of an effective control of gastro-oesophageal reflux in reflux patients is still lacking. PMID- 11856075 TI - Budesonide enema in pouchitis--a double-blind, double-dummy, controlled trial. AB - BACKGROUND: Pouchitis has been suggested to be a recurrence of ulcerative colitis in a colon-like mucosa. Topical steroids are a valid therapeutic alternative for distal forms of ulcerative colitis. AIM: To investigate the efficacy and tolerability of budesonide enema in the treatment of pouchitis compared with oral metronidazole. MATERIALS AND METHODS: Twenty-six patients with an active episode of pouchitis (defined as a pouchitis disease activity index score >or= 7) and no treatment during the previous month were randomized to receive either budesonide enema (2 mg/100 mL at bedtime) plus placebo tablets or oral metronidazole (0.5 g b.d.) plus placebo enema in a prospective, double-blind, double-dummy, 6-week, controlled trial. RESULTS: Based on the intention-to-treat principle, we detected a significant improvement in disease activity at the end of the first week with both drugs (P < 0.01). After that, improvement was moderated until stabilization at 4 weeks in both treatments. The per protocol analysis showed that both drugs had similar efficacy in terms of disease activity, clinical and endoscopic findings. Fifty-eight per cent and 50% of patients improved (decrease in pouchitis disease activity index >or= 3) with budesonide enema and metronidazole, respectively (odds ratio, 1.4; confidence interval, 0.2-8.9). Adverse effects were observed in 57% of patients given metronidazole and in 25% of patients given budesonide. CONCLUSIONS: Budesonide enemas are an alternative treatment for active pouchitis, with similar efficacy but better tolerability than oral metronidazole. PMID- 11856076 TI - Lansoprazole vs. omeprazole for gastro-oesophageal reflux disease: a pH-metric comparison. AB - BACKGROUND: Lansoprazole and omeprazole are widely used proton pump inhibitors for the management of gastro-oesophageal reflux. Normalization of oesophageal acid exposure is an important goal in the management of complicated and atypical gastro-oesophageal reflux disease. AIM: To compare the efficacy of lansoprazole and omeprazole in the abolition of abnormal reflux as assessed by oesophageal pH monitoring. METHODS: Seventy patients with complicated or atypical gastro oesophageal reflux disease were randomly assigned to receive 30 mg lansoprazole or 20 mg omeprazole once daily. Three to four weeks after the start of treatment, patients underwent oesophageal pH monitoring while on therapy. If the results were still abnormal, the proton pump inhibitor dosage was doubled and 24-h pH metry was repeated after 20-30 days. RESULTS: Thirty-six patients were randomized to receive lansoprazole and 34 patients to receive omeprazole. Ten of the 36 (29%) patients treated with 30 mg lansoprazole once daily and 23 of the 34 (68%) patients treated with 20 mg omeprazole once daily had persistently abnormal reflux at oesophageal pH monitoring (P < 0.001). In all such cases, repeat pH monitoring after doubling the proton pump inhibitor dosage gave normal results. CONCLUSIONS: At the currently marketed dosages of lansoprazole and omeprazole, normalization of oesophageal acid exposure in patients is accomplished more easily with lansoprazole. PMID- 11856077 TI - Medical decision analysis of endoscopic surveillance of Barrett's oesophagus to prevent oesophageal adenocarcinoma. AB - BACKGROUND: Barrett's oesophagus is associated with an increased risk of the development of oesophageal adenocarcinoma. Endoscopic surveillance every 2-5 years has been recommended to prevent death from adenocarcinoma. AIM: To assess the cost-effectiveness of this strategy. METHODS: The incremental cost effectiveness of surveillance (as compared to no surveillance) was analysed with a computer model of a Markov process. RESULTS: Compared to no surveillance, the incremental cost-effectiveness of bi-annual endoscopy is 16,695 dollars per life year saved. Surveillance is less cost-effective if the incidence rate of oesophageal adenocarcinoma is low and the 5-year survival rate is high. For surveillance to be cost-effective, there should be little reduction in health related quality of life following surgical oesophagectomy to prevent death. Moreover, endoscopic surveillance and oesophagectomy need to be efficacious in reliably diagnosing high-grade dysplasia and preventing deaths from cancer. If such ideal conditions of surveillance are not met, the cost per life-year saved could rise five-fold. CONCLUSIONS: Endoscopic surveillance of patients with Barrett's oesophagus may be a cost-effective means to prevent death from oesophageal adenocarcinoma. PMID- 11856078 TI - The epidemiology and natural history of Crohn's disease in population-based patient cohorts from North America: a systematic review. AB - AIM: To quantify, through systematic review, the epidemiology and natural history of Crohn's disease in North America. METHODS: The selected articles contained: (i) population-based samples of patients followed from the time of diagnosis; and (ii) objective diagnostic criteria for disease. Studies on the natural history of Crohn's disease also contained sufficient follow-up. DATA COLLECTION AND ANALYSIS: For prevalence studies, data on the incidence, prevalence, gender and age at diagnosis were extracted. For natural history studies, data on the disease activity, use of medications and surgery were extracted. MAIN RESULTS: The prevalence of Crohn's disease in North America ranges from 26.0 to 198.5 cases per 100,000 persons. The incidence rates range from 3.1 to 14.6 cases per 100,000 person-years. Most patients have a chronic intermittent disease course, while 13% have an unremitting disease course and 10% have a prolonged remission. Less than half require corticosteroids at any point. During any given year, approximately 10% are treated with corticosteroids and 30% are treated with 5-aminosalicylates. Up to 57% of patients require at least one surgical resection. CONCLUSIONS: Between 400,000 and 600,000 patients in North America have Crohn's disease, and the natural history is marked by frequent exacerbations requiring treatment with corticosteroids, 5-aminosalicylate products and surgery. PMID- 11856079 TI - A double-blind comparison of balsalazide, 6.75 g daily, and sulfasalazine, 3 g daily, in patients with newly diagnosed or relapsed active ulcerative colitis. AB - BACKGROUND: Sulfasalazine is well established in the treatment of active ulcerative colitis. Intolerance to sulfasalazine, however, is a common problem. Balsalazide has been designed to deliver 5-aminosalicylic acid to the colon without the poor tolerability of sulfasalazine. AIM: To compare the safety and efficacy of balsalazide, 6.75 g daily, with sulfasalazine, 3 g daily, in the treatment of active ulcerative colitis of all grades of severity. METHODS: Balsalazide and sulfasalazine were compared in a multicentre, double-blind, parallel group study over 12 weeks. Patients were stratified for disease severity and topical and/or oral steroids were co-administered where clinically necessary. RESULTS: Fifty-seven patients were randomized: 28 to receive balsalazide and 29 to receive sulfasalazine. Significantly fewer patients withdrew from the balsalazide group due to adverse events (2/28 vs. 9/29, P=0.041). These data confirm that balsalazide is better tolerated than sulfasalazine. In patients able to tolerate the treatment, similar improvements were recorded in clinical, sigmoidoscopic and histological assessments in both treatment groups. CONCLUSIONS: This study confirms the better tolerability of balsalazide compared to sulfasalazine, and supports the use of balsalazide in ulcerative colitis of all grades of severity. PMID- 11856081 TI - Interaction between azathioprine and aminosalicylates: an in vivo study in patients with Crohn's disease. AB - BACKGROUND: The inhibition of thiopurine methyltransferase activity, one of the enzymes responsible for azathioprine metabolism, by aminosalicylates has been described in an in vitro study. This could result in a higher risk of bone marrow depression when using the two drugs together. AIM: To investigate the in vivo interaction between azathioprine and aminosalicylates in quiescent Crohn's disease by measuring 6-thioguanine nucleotide levels, thiopurine methyltransferase activity and the plasma levels of the acetylated metabolite of 5-aminosalicylic acid. METHODS: Sixteen patients taking a stable dose of azathioprine, plus sulfasalazine or mesalazine, were enrolled and completed the study. They were not taking any drugs interfering with azathioprine metabolism. Four visits every 4 weeks were held over a 3-month period. Aminosalicylate administration was withdrawn after the second visit. At each visit, the blood cell count, inflammatory parameters, levels of 6-thioguanine nucleotide and the acetylated metabolite of 5-aminosalicylic acid and thiopurine methyltransferase activity were determined. RESULTS: After aminosalicylate withdrawal, mean 6 thioguanine nucleotide levels decreased significantly from 148 pmol (57-357 pmol) to 132 pmol (56-247 pmol) per 8 x 10(8) red blood cells (P=0.027), without significant changes in thiopurine methyltransferase activity or biological parameters. CONCLUSIONS: This in vivo study favours the existence of an interaction between azathioprine and aminosalicylates through a mechanism which remains unclear. This drug-drug interaction should be taken into account when using azathioprine and aminosalicylates simultaneously. PMID- 11856080 TI - A double-blind comparison of balsalazide, 6.75 g, and sulfasalazine, 3 g, as sole therapy in the management of ulcerative colitis. AB - BACKGROUND: Sulfasalazine is accepted therapy for active ulcerative colitis, but side-effects and intolerance are common. Balsalazide is an azo-bonded pro-drug which also releases 5-aminosalicylic acid into the colon, but uses an inert carrier molecule. AIM: To compare the safety and efficacy of sul- fasalazine, 3 g, with balsalazide, 6.75 g, in the initial daily treatment of mild to moderate ulcerative colitis. METHODS: A randomized, multicentre, double-blind, parallel group study was performed, with a treatment duration of 8 weeks. Patients on previous maintenance treatment were excluded. The trial medication was the sole treatment for the colitis. Efficacy was assessed by patient diaries, symptom assessment, sigmoidoscopic appearance and histology. RESULTS: Fifty patients were recruited: 26 allocated to the balsalazide group and 24 to the sulfasalazine group. More patients withdrew due to adverse events in the sulfasalazine group (nine patients vs. one patient in the balsalazide group, P=0.004). Improvement occurred in both groups, with a tendency to a faster response with balsalazide. Of the patients taking balsalazide, 61% achieved clinical and sigmoidoscopic remission. CONCLUSIONS: Balsalazide, 6.75 g, is effective as the sole treatment for patients with mild to moderately active ulcerative colitis, with significantly fewer withdrawals due to side-effects than in a similar group of patients taking sulfasalazine, 3 g. PMID- 11856082 TI - Efficacy and safety of oral ridogrel in the treatment of ulcerative colitis: two multicentre, randomized, double-blind studies. AB - BACKGROUND: Ridogrel at low doses inhibits thromboxane synthase. Oral ridogrel, from 5 mg once daily to 150 mg twice daily, improves the endoscopic appearance of colonic mucosa and clinical manifestations in mild to moderate ulcerative colitis. AIM: One US trial and one international trial were conducted to determine the effect of ridogrel on mild to severe active ulcerative colitis. METHODS: Two 12-week, double-blind, randomized, parallel-group trials were conducted. A US trial compared 0.5 mg, 2.5 mg and 5 mg of ridogrel once daily with placebo. An international trial compared 0.5 mg of ridogrel once daily with 2.5 mg and 5.0 mg of ridogrel once daily and 800 mg of mesalazine (known as mesalamine in the USA) three times daily. The primary efficacy outcome measure was the rate of complete remission. RESULTS: In the US trial, complete remission was achieved in 20.8% of patients in the 0.5 mg ridogrel group, 17.9% in the 2.5 mg ridogrel group, 20.6% in the 5.0 mg ridogrel group and 13.6% in the placebo group. In the international trial, 14.4% of patients in the 0.5 mg ridogrel group, 19.6% in the 2.5 mg ridogrel group, 19.4% in the 5.0 mg ridogrel group and 16.4% in the mesalazine group experienced complete remission. In the international trial, rates of complete remission at the end-point were greater in the 2.5 mg and 5.0 mg ridogrel groups than in the 0.5 mg ridogrel group, but the differences were not statistically significant. In the US trial, rates of complete remission at the end-point were greater in the 2.5 mg and 5.0 mg ridogrel groups than in the placebo group, but the differences were not statistically significant. Approximately 30% of the patients in each group discontinued treatment before the 12-week end-point owing to a lack of therapeutic response. All doses of ridogrel were well tolerated and comparable with placebo or mesalazine in terms of safety. CONCLUSIONS: No significant differences in the primary efficacy outcome measure were found between either the 2.5 mg or the 5.0 mg dose of ridogrel and placebo in the US trial and between either the 2.5 mg or the 5.0 mg dose of ridogrel and the 0.5 mg dose of ridogrel, a surrogate dose for placebo, in the international trial. There was no clear indication in either trial of an effective dose of ridogrel in the treatment of ulcerative colitis. PMID- 11856083 TI - Topical mononitrate treatment in patients with anal fissure. AB - AIM: To assess the efficacy and patient compliance of topical mononitrate hydrogel for the treatment of anal fissure. MATERIALS AND METHODS: Nineteen patients with symptomatic chronic anal fissures were randomly allocated to receive either active (10 patients) or placebo (nine patients) gel treatment. Rectal administration of hydrogel containing 0.2% isosorbide-5-mononitrate was prescribed. Patients were instructed on its application to the anal canal twice daily for 3 weeks. A questionnaire was used to determine patient compliance with therapy. Anal manometry was performed before and after therapy. RESULTS: At the end of therapy, the fissures were healed in 80% of actively treated patients compared with 22% of the control group. There was a mean reduction of 28% in mean resting anal pressure. Two actively treated patients (20%) suffered from mild headache relieved with oral analgesics and menthol lozenges. Faecal incontinence was not observed. There were no recurrences during at least 3 months of follow up. CONCLUSIONS: Topical mononitrate gel therapy of anal fissures is an effective and safe approach. In this study, the few cases of headache were rapidly relieved with oral analgesia and menthol lozenges. PMID- 11856084 TI - Appropriateness and diagnostic yield of upper gastrointestinal endoscopy in an open-access endoscopy system: a prospective observational study based on the Maastricht guidelines. AB - AIM: To test the appropriateness of referrals for upper gastrointestinal endoscopy in Campania, Italy, using the criteria of the Maastricht Consensus. PATIENTS: Patients undergoing endoscopy during a 1-week period in 21 Endoscopy Services were considered prospectively. The reasons for endoscopy were dyspeptic symptoms, history of peptic ulcer and assessment after treatment. The age, sex, symptoms, history of peptic ulcer (previous endoscopic or radiographic examinations and treatment), endoscopic diagnosis and H. pylori status were recorded. The indications for endoscopy were evaluated according to the Maastricht guidelines. RESULTS: Two hundred and sixteen of 706 patients presented with reflux symptoms, 430 with dyspeptic symptoms, 38 with alarm symptoms and 22 with atypical symptoms. Endoscopy was normal in 376 cases (53.2%); duodenal ulcer was found in 219, gastric ulcer in 45, oesophagitis in 82 and gastric cancer in six. All patients with cancer were older than 45 years, and four presented with alarm symptoms. In 398 cases (56%), endoscopy was considered not to be indicated: 250 patients with a previous diagnosis of ulcer without a change in symptoms, 38 patients in order to confirm eradication and 110 patients younger than 45 years with dyspepsia without alarm symptoms. CONCLUSIONS: A large number of endoscopic examinations could be avoided by following the guidelines of the Maastricht Consensus. In order to reduce endoscopic workload, strategies for educating physicians should be pursued. PMID- 11856085 TI - Safety and efficacy of glucagon as a premedication for upper gastrointestinal endoscopy--a comparative study with butyl scopolamine bromide. AB - BACKGROUND: Glucagon inhibits digestive motility and is used for endoscopic premedication; however, its effect on cardiopulmonary function during endoscopy has not yet been fully investigated. AIM: To clarify the efficacy and safety of glucagon compared with butyl scopolamine bromide as upper gastrointestinal endoscopy premedication. METHODS: Two hundred and forty consecutive patients over 40 years of age, referred for upper gastrointestinal endoscopy, without any complications, were studied. These patients were randomly premedicated with butyl scopolamine bromide (SC group) or glucagon (G group). Time course changes in blood pressure, arterial oxygen saturation, heart rate and the number of retching episodes during endoscopy were examined. The efficacy of glucose tablets after upper gastrointestinal endoscopy to prevent hypoglycaemia caused by glucagon was evaluated. Cardiopulmonary parameters were also examined in 77 complicated patients with glucagon premedication (GC group). RESULTS: A continuous increase in heart rate during upper gastrointestinal endoscopy was observed in the SC group, but not in the G and GC groups. Blood pressure, arterial oxygen saturation and number of retching episodes were not different between the groups. Hypoglycaemia-related symptoms were frequent in the G group without glucose tablets, but were prevented by the administration of glucose. CONCLUSIONS: Glucagon has a weaker effect on cardiopulmonary function during upper gastrointestinal endoscopy than butyl scopolamine bromide. Glucose administration prevents hypoglycaemia-related symptoms caused by glucagon. PMID- 11856086 TI - Does intravenous ondansetron affect gastric emptying of a solid meal, gastric electrical activity or blood hormone levels in healthy volunteers? AB - BACKGROUND: In previous studies, tropisetron has been shown to accelerate gastric emptying of a solid meal. However, it is uncertain whether other specific 5 hydroxytryptamine-3 receptor antagonists, such as ondansetron, also have a gastroprokinetic effect in humans. AIM: To evaluate the effect of ondansetron on gastric half-emptying time (T1/2) of a solid meal, gastric myoelectrical activity and hormone levels in 14 healthy volunteers. METHODS: In a placebo-controlled, randomized, crossover study, we investigated the effects of ondansetron (8 mg intravenously) on the gastric emptying of solids (by scintigraphy), gastric myoelectrical activity (by electrogastrography) and the post-prandial release of cholecystokinin, gastrin, human pancreatic polypeptide, gastric inhibitory polypeptide, vasoactive intestinal polypeptide, motilin, substance P and galanin. RESULTS: The average T1/2 values were 86 min and 85.5 min without lag time (P=0.082) and 92 min and 93 min with lag time (P=0.158) for the placebo and ondansetron treatments, respectively. The average T1/2 of female volunteers was significantly longer than that of male volunteers. The dominant gastric electrical frequency and hormone plasma concentrations were not altered by ondansetron. CONCLUSIONS: Ondansetron did not affect the gastric emptying of solids, the dominant gastric electrical frequency or the plasma concentrations of the analysed gastrointestinal peptides. PMID- 11856087 TI - Dose-related effects of motilin on proximal gastrointestinal motility. AB - AIM: To assess non-invasively the dose-response relations for the effects of exogenous motilin on antrum contraction frequency, gall-bladder volume and gastric myoelectrical activity. METHODS: In a double-blind, randomized, placebo controlled, five-way crossover study, 10 fasted healthy volunteers were infused intravenously with synthetic human motilin (0.5, 1, 2 and 4 pmol x min/kg) or placebo for 60 min. Gall-bladder volume and antrum contractions were assessed by ultrasonography and gastric myoelectrical activity by electrogastrography. Motilin concentrations were measured using a radioimmunoassay. RESULTS: Baseline plasma motilin levels (60 pmol/L) were similar for all treatments. Motilin levels increased upon the start of infusion and rapidly returned to baseline after cessation of the infusion. At motilin doses of 2 and 4 pmol.min/kg, the antrum contraction frequency was significantly augmented, with maximum differences of two contractions per 2-min interval compared to placebo, while no changes in gastric myoelectrical activity were observed. Changes in gall-bladder volume were not significantly different for any of the motilin doses compared to placebo. CONCLUSIONS: Motilin increased antrum contraction frequency, whereas no effect on gastric myoelectrical activity was observed. Antrum contraction frequency appears to be a useful biomarker for motilin efficacy, and motilin doses of 2 and 4 pmol x min/kg were equally effective. PMID- 11856088 TI - Impact of intravenous omeprazole on Helicobacter pylori eradication by triple therapy in patients with peptic ulcer bleeding. AB - AIM: To test the impact of intravenous omeprazole on Helicobacter pylori eradication for bleeding peptic ulcers. METHODS: A total of 175 H. pylori infected patients with bleeding peptic ulcers were randomized into either an omeprazole group or a ranitidine group, receiving intravenous omeprazole or ranitidine for 3 days after endoscopy. Afterwards, 1-week triple therapy was used to eradicate H. pylori for both groups. Six weeks later, either a 13C-urea breath test or follow-up endoscopy was performed to assess the success of H. pylori eradication. RESULTS: The rebleeding rate was lower in the omeprazole group vs. the ranitidine group (6% vs. 17%, P < 0.05). The H. pylori eradication rate was higher in the omeprazole group (intention-to-treat analysis: 83% vs. 66%, P < 0.05; per protocol analysis: 93% vs. 80%, P < 0.05). For patients with duodenal ulcers, the per protocol H. pylori eradication rate of the omeprazole group was higher than that of the ranitidine group (93% vs. 73%, P < 0.05). CONCLUSIONS: Intravenous omeprazole can decrease the risk of rebleeding of peptic ulcers. For duodenal ulcers, in particular, intravenous omeprazole may even improve the H. pylori eradication rate of the subsequent triple therapy. PMID- 11856089 TI - Efficacy of low-dose clarithromycin triple therapy and tinidazole-containing triple therapy for Helicobacter pylori eradication. AB - BACKGROUND: Proton pump inhibitor-based triple therapies are recommended as the first-line treatment for Helicobacter pylori eradication. AIM: To evaluate the efficacies of low-dose clarithromycin triple therapy and tinidazole-containing triple therapy in a metronidazole resistance prevalent area and to compare the efficacies with standard triple therapy. METHODS: In a randomized, multicentre, prospective study, a total of 352 patients with duodenal ulcer or non-ulcer dyspepsia were randomly divided into three groups according to the administered regimen: OAC250 group (omeprazole, 20 mg, amoxicillin, 1000 mg, and clarithromycin, 250 mg), OAC500 group (omeprazole, 20 mg, amoxicillin, 1000 mg, and clarithromycin, 500 mg) and OTC group (omeprazole, 20 mg, tinidazole, 500 mg, and clarithromycin, 500 mg). The three groups received each regimen twice daily for 7 days. Upper gastrointestinal endoscopy was performed before and 4 weeks after treatment. H. pylori status was determined by rapid urease test and 13C urea breath test. RESULTS: The eradication rates in the OAC250, OAC500 and OTC groups were 76.2%, 65.7% and 64.8% (95% confidence interval: 67.9-84.4%, 56.7 74.8% and 55.7-73.9%), respectively, by intention-to-treat analysis (P=0.149) and 92.8%, 87.2% and 84.1% (95% confidence interval: 84.4-97.3%, 77.9-93.8% and 73.9 91.2%), respectively, by per protocol analysis (P=0.088). All regimens were well tolerated and compliance was excellent. CONCLUSIONS: Both low-dose clarithromycin triple therapy and tinidazole-containing triple therapy are effective and safe regimens for H. pylori eradication. PMID- 11856090 TI - Low-dose lansoprazole and clarithromycin plus metronidazole vs. full-dose lansoprazole and clarithromycin plus amoxicillin for eradication of Helicobacter pylori infection. AB - AIM: To compare, in a randomized controlled trial, the efficacy and tolerability of two 1-week triple therapies for Helicobacter pylori eradication. METHODS: One hundred and thirty-four consecutive patients with non-ulcer dyspepsia and H. pylori infection were randomized to receive lansoprazole 30 mg once daily, clarithromycin 250 mg twice daily, and metronidazole 500 mg twice daily (LCM group), or lansoprazole 30 mg twice daily, clarithromycin 500 mg twice daily, and amoxicillin 1000 mg twice daily (LCA group). H. pylori status was assessed by rapid urease test, histology and 13C-urea breath test before and after therapy. RESULTS: At 3 months, H. pylori eradication (intention- to-treat/per protocol analysis) was 92.4%/93.8% in the LCM group and 83.1%/85.7% in the LCA group (P=N.S.). Side-effects were more frequently reported in the LCA group (37.9%) than in the LCM group (19.7%) (P < 0.05). CONCLUSIONS: In this open, randomized controlled trial, eradication of H. pylori by low-dose lansoprazole and clarithromycin plus metronidazole was higher with significantly less side-effects than by full-dose lansoprazole and clarithromycin plus amoxicillin. This finding may be related to the stronger synergism of clarithromycin plus metronidazole, even at lower doses, than of clarithromycin plus amoxicillin. Considering the lower cost as well, LCM should be preferred to LCA in the eradication of H. pylori. PMID- 11856092 TI - Laparoscopy in urology: indications and training. PMID- 11856091 TI - Treatment with a proton pump inhibitor promotes corpus gastritis in patients with Helicobacter pylori-infected antrum-predominant gastritis. AB - BACKGROUND: Proton pump inhibitors have been reported to modify the level of Helicobacter pylori gastritis. AIM: To quantitatively investigate the effect of a proton pump inhibitor on the mucosal neutrophil reaction. METHODS: Forty-six H. pylori-infected patients (17 duodenal ulcer, 29 gastric ulcer) were enrolled. During endoscopic examination, biopsy samples were obtained from the antrum and the corpus. The tissue content of neutrophil myeloperoxidase was measured by enzyme-linked immunoabsorbent assay, and H. pylori infection was histologically assessed. A proton pump inhibitor was administered orally for 8 weeks. RESULTS: In the patients as a whole, antral myeloperoxidase decreased significantly after proton pump inhibitor treatment, but corpus myeloperoxidase remained largely unchanged. In duodenal ulcer patients, myeloperoxidase significantly decreased in the antrum, but increased in the corpus. In gastric ulcer patients, a significant reduction was observed in antral myeloperoxidase, but corpus myeloperoxidase remained unchanged. In the antral myeloperoxidase > corpus myeloperoxidase subgroup (n=24), antral myeloperoxidase significantly decreased, whereas corpus myeloperoxidase increased. No changes were observed at either site in the corpus myeloperoxidase > antral myeloperoxidase subgroup. Histology showed that the antral bacterial load of H. pylori decreased in all subgroups, but that it was mostly unchanged in the corpus. CONCLUSIONS: Proton pump inhibitor treatment stimulated the neutrophil reaction in the corpus mucosa of duodenal ulcer patients and of patients in whom antral neutrophil accumulation was more predominant than that of the corpus. This phenomenon may not be caused by increased bacterial density. PMID- 11856093 TI - The URO Mentor: development and evaluation of a new computer-based interactive training system for virtual life-like simulation of diagnostic and therapeutic endourological procedures. AB - OBJECTIVE: To overcome the current disadvantages of traditional training methods for ureterorenoscopy and percutaneous nephrolitholapaxy, using the URO Mentor (Simbionix, Tel Aviv, Israel) computer-assisted simulator. METHODS: The URO Mentor device for training and quality control in ureterorenoscopy was developed using virtual reality, multimedia technology and intelligent tutoring systems. The central software system features a proprietary visualization engine (the SVE) which allows real-time simulation by offering a high-level object-orientated application program interface (written in C++) available for use with either Microsoft, DirectX 7 or OpenGL as platforms. The SVE includes general procedures to allow two- (2D) and three-dimensional (3D) rendering, collision detection, collision correction, 3D morphing, 2D image manipulation, texture mapping, 'bump' mapping, video texture, X-ray rendering, special effects (blood, smoke, stone fragments and more) and reflections. RESULTS: The system allows a complete training session on a wide range of procedures by offering different types of cases and virtual patients, and features a full representation of the endourological procedures under direct vision and by using interactive fluoroscopy with a contrast agent. The supported tools include: baskets, graspers, intracorporal lithotripters, guidewires, catheters, stents, biopsy and dilatation devices. The endourological procedures that can be performed are lithotripsy, tumour resection, treatment of strictures and obstructions, stent placement and biopsies. CONCLUSION: The URO Mentor introduces a new generation of mannequin equipped with a special haptic device, providing trainees with an unparalleled life-like sensation while training for diagnostic and therapeutic endourological procedures. By bringing key advances into urological simulation (e.g. with the real-time X-ray renderer) and by integrating in a single system both high-quality simulation and learning tools, the URO Mentor provides new perspectives for computer-based urological training systems and methods. PMID- 11856094 TI - Coding errors: a comparative analysis of hospital and prospectively collected departmental data. AB - OBJECTIVES: To assess the accuracy of a hospital coding database at a busy tertiary referral urological unit. METHODS: Prospectively collected departmental coding data for all urological patients attending the Churchill Hospital, Oxford between 1 May 1999 and 30 April 2000 were compared with the coding data entered by hospital coding clerks on the Oxford Radcliffe NHS Trust database. RESULTS: There were significant discrepancies between the number of patients on the hospital and the departmental database (639 vs 1109). There were gross procedural coding errors in 74 cases. CONCLUSION: Hospital-coded data in this study were incomplete and inaccurate. This has important implications when considering the validity of hospital-trust databases when used as a source for medical research, clinical audit and as a representation of a consultants' clinical workload. PMID- 11856096 TI - Bladder regeneration: between the idea and reality. PMID- 11856095 TI - Day-case ureteroscopy: an observational study. AB - OBJECTIVE: To evaluate the outcome after day-case ureteroscopy (used in diagnosing and managing ureteric disease, primarily urolithiasis), as awareness of reduced resources has resulted in increasing pressure to undertake procedures in a day-surgery setting. PATIENTS AND METHODS: All patients presenting to the unit and requiring ureteroscopy between May 1995 and May 2000 were considered for a day-surgery procedure. The assessment of suitability comprised anaesthetic and social factors; no urological criteria precluded a day-surgery procedure. Outcomes after day-case ureteroscopy, including immediate or delayed admissions and subsequent inpatient management, were reviewed retrospectively. RESULTS: Sixty-three day-case ureteroscopies were performed on 56 patients (mean age 47 years, range 19-78); eight procedures were diagnostic. Therapeutic ureteroscopies included one balloon dilatation of a ureteric stricture and 54 procedures for urolithiasis, with 98% stone clearance. Most patients were discharged with a JJ stent in situ. Of nine patients requiring immediate admission, seven were for pain control; eight were discharged on the following day. Seven patients required delayed admission 1-13 days after the procedure, three for stent-related symptoms and three for infection. No significant predictors of immediate or delayed admission were identified, although antibiotic prophylaxis was associated with a reduced admission rate. CONCLUSION: Ureteroscopy can be used successfully as a planned day-case procedure in a dedicated day-surgery unit, with few patients requiring hospitalization. Implementation of analgesia protocols and routine antibiotic prophylaxis may reduce admission after day-case ureteroscopy. PMID- 11856097 TI - A new small-calibre diagnostic flexible cystoscope. AB - OBJECTIVE: To determine whether flexible cystoscopy with a prototype slim flexible instrument is less uncomfortable than flexible cystoscopy using a standard flexible cystoscope. PATIENTS AND METHODS: In all, 200 men scheduled to undergo flexible cystoscopy for the follow-up of transitional cell carcinoma of the bladder were randomized in equal groups to flexible cystoscopy using the standard instrument or using the prototype slim-scope. Each patient had 11 mL of 2% lignocaine gel instilled intraurethrally 5 min before the procedure. At the end of the procedure the patient was asked to complete a 100-mm non-graphical visual analogue scale to rate how uncomfortable the procedure was. RESULTS: The slim-scope was very significantly less uncomfortable than the conventional instrument (anova P < 0.001). The ease of use and views of the bladder were similar to those obtained with the conventional cystoscope. CONCLUSION: The slim scope offers a significant benefit in patient comfort, with little loss of image quality or ease of use. PMID- 11856098 TI - Thick vs thin loop transurethral resection of the prostate: a double-blind prospective trial of early morbidity. AB - OBJECTIVE: To compare blood loss, irrigation requirements and hospital stay between standard transurethral resection of the prostate (TURP) and resection incorporating vaporization, i.e. transurethral vaporization resection (TUVRP). PATIENTS AND METHODS: Seventy patients were prospectively randomized in a blinded fashion between TURP with a standard resection loop (thin loop) or resection with a roller cutting electrode (thick loop). The outcome was assessed as the change in haemoglobin at 30 min and 24 h after the resection finished. Secondary outcome measures were irrigation requirements, length of catheterization, hospital stay, changes in serum sodium, and complications. The nursing staff and surgical registrar managing the patients after surgery were unaware of the resection technique used. RESULTS: Data were available for 65 patients (36 thin loop and 29 thick loop resections). The mean decrease in haemoglobin after TURP was 14.8 g/L at 30 min and 11.8 g/L at 24 h; for TUVRP the corresponding values were 14.6 and 14.2 g/L (P = 0.69). In addition, there was no difference in irrigation requirements, changes in serum sodium, length of catheterization, hospital stay or bleeding complications. CONCLUSION: The use of a thick loop at higher cutting current settings during TUVRP offers no advantage over standard thin-loop TURP. PMID- 11856099 TI - Detecting and quantifying absorbed irrigation fluid by measuring mannitol and sorbitol concentrations in serum samples, and by ethanol monitoring. AB - OBJECTIVE: To describe a modified and improved technique which, in one measurement, estimates the influx of irrigation fluid during endoscopic endometrial ablation or prostate resection, and provides both rapid confirmation of the diagnosis and an estimate of the amount of fluid absorbed by detecting markers which pass from the irrigation fluid to the serum, i.e. mannitol or sorbitol. PATIENTS AND METHODS: Control samples were taken for analysis before irrigation, and test samples were taken on four occasions during and after intervention, from each of 10 patients undergoing transurethral resection of the prostate. Irrigation fluid was also marked with ethanol (1.5% w/v) and the concentration of this agent measured in the blood and expired air of these patients. The absorbed volume was calculated according to the extracellular distribution space of mannitol. RESULTS: Mannitol and sorbitol could be measured in 85% and 73% of the 40 test samples, respectively. The threshold for full sensitivity for breath ethanol concentration to detect absorption was 132 mL. CONCLUSION: This method for detecting serum mannitol and sorbitol represents a valid procedure for confirming and quantifying the absorption of irrigation fluid in the clinic, which agrees closely with the already established ethanol monitoring procedure and which should now be considered as a reference procedure. PMID- 11856100 TI - Sexual function before and after various treatments for symptomatic benign prostatic hyperplasia. AB - OBJECTIVE: To determine the effect of various treatments for benign prostatic hyperplasia (BPH) on sexual function. PATIENTS AND METHODS: In a longitudinal multicentre study carried out in the Netherlands, 670 consecutive patients with BPH (aged > or = 50 years) were examined at baseline and 9 months after inclusion. All patients completed a questionnaire about symptomatology, bothersomeness and sexual function before and after treatment. Other diagnostic information was retrieved from the medical records. RESULTS: At baseline, 49-59% of the population (mean age 66 years) reported normal potency and 69% a normal libido. Outcomes at 9 months were stratified by type of treatment, i.e. surgery in 207, alpha-blockers in 43, finasteride in 47 and watchful waiting in 234 men. Patients with multiple or other treatments (131) or recent surgery (eight) were excluded from the analyses. For the four sexual items included, 84% of the patients reported no changes. All treatments showed both improvement and deterioration in 3-14% and 0-16% of the patients, respectively. CONCLUSION: For sexual function, most patients remained stable after treatment for BPH, while positive and negative changes occurred in equal proportions for all treatments. These results question the previously reported high incidence of sexual adverse events, particularly after surgical intervention. PMID- 11856101 TI - Terazosin for treating symptomatic benign prostatic obstruction: a systematic review of efficacy and adverse effects. AB - OBJECTIVE: To systematically review and evaluate the effectiveness and adverse effects of the alpha-antagonist, terazosin, for treating urinary symptoms associated with benign prostatic obstruction (BPO). METHODS: Studies were sought and included in the review if they were randomized trials of at least 1 month duration, involved men with symptomatic BPO and compared terazosin with placebo or active controls. The study, patient characteristics and outcome data were extracted in duplicate onto standardized forms using a prospectively developed protocol. RESULTS: Seventeen studies involving 5151 men met the inclusion criteria, i.e. placebo-controlled (10), alpha-blockers (seven), finasteride alone or combined with terazosin and placebo (one), and microwave therapy (one). The study duration was 4-52 weeks; the mean age of the men was 65 years and 82% were white. Baseline urological symptom scale scores and flow rates showed that men had moderate BPO. Efficacy outcomes were rarely reported in a way that allowed for data pooling, but indicated that terazosin improved symptom scores and flow rates more than did placebo or finasteride, and similarly to other alpha antagonists. The pooled mean percentage improvement for the Boyarsky symptom score was 37% for terazosin and 15% for placebo (four studies). The mean percentage improvement for the American Urological Association symptom score was 38%, compared with 17% and 20% for placebo and finasteride, respectively (two studies). The pooled mean improvement in the International Prostate Symptom Score of 40% was similar to that with tamsulosin (43%). Peak urinary flow rates improved more with terazosin (22%) than with placebo (11%) and finasteride (15%), but did not differ significantly from the other alpha-antagonists. The percentage of men discontinuing terazosin was comparable with those receiving placebo and finasteride, but greater than with other alpha-antagonists. Adverse effects were greater than with placebo and included dizziness, asthenia, headache and postural hypotension. CONCLUSIONS: The available evidence indicates that terazosin improves the symptoms and flow rates associated with BPO; it was more effective than placebo or finasteride and similar to other alpha-antagonists. Adverse effects were generally mild but more frequent than with other alpha-antagonists and associated with a two- to four-fold increase in treatment discontinuation. PMID- 11856102 TI - Transurethral needle ablation for chronic nonbacterial prostatitis. AB - OBJECTIVE: To investigate the efficacy of transurethral needle ablation (TUNA) of the prostate for treating chronic nonbacterial prostatitis unresponsive to conservative therapies. PATIENTS AND METHODS: Forty-two patients (mean age 38.5 years, range 25-52) with nonbacterial prostatitis in whom clinical management was unsuccessful in relieving the symptoms or signs of prostatitis were treated using TUNA. All patients had a high leukocyte count (> 15 per high-power field) in expressed prostatic secretions (EPS) with no bacterial growth in either urine or prostatic secretion cultures. Before TUNA all patients were evaluated using a symptom score, satisfaction score (quality of life) and an examination of prostatic secretions. All patients were reassessed using the same variables 1 and 3 months after TUNA. Of the 42 patients, 10 had their semen analysed before and 3 months after treatment. RESULTS: The mean (sd) symptom and satisfaction scores improved significantly, from 11.02 (2.90) to 5.00 (2.61) and from 4.84 (0.57) to 1.26 (1.18), respectively, 3 months after TUNA (both P < 0.05). Of the 42 patients, 30 (71%) had normal EPS results within 3 months of TUNA. Also, of 37 patients with high leukocyte counts (> 100 per high-power field) before TUNA, 26 (70%) had normal EPS results within 3 months afterward, and 29 (78%) had a satisfaction score of < 3 points. CONCLUSIONS: TUNA may be a possible treatment option for patients with chronic nonbacterial prostatitis that is unresponsive to conservative therapies. A long-term follow-up and randomized studies are needed to confirm the efficacy of the TUNA against intractable chronic prostatitis. PMID- 11856103 TI - Growth factors and their receptors in prostate cancer. PMID- 11856104 TI - Prostate brachytherapy has come of age: a review of the technique and results. PMID- 11856105 TI - Can diet affect prostate cancer? PMID- 11856106 TI - Making decisions about treatment for localized prostate cancer. AB - OBJECTIVE: To describe the decision-making processes used by men diagnosed with localized prostate cancer who were considering treatment. PATIENTS AND METHODS: Men newly diagnosed with localized prostate cancer from outpatient urology clinics and urologists' private practices were approached before treatment. Their decision-making processes and information-seeking behaviour was assessed; demographic information was also obtained. RESULTS: Of 119 men approached, 108 (90%) were interviewed; 91% reported non-systematic decision processes, with deferral to the doctor, positive and negative recollections of others' cancer experiences, and the pre-existing belief that surgery is a better cancer treatment being most common. For systematic information processing the mean (sd, range) number of items considered was 4.19 (2.28, 0-11), with 57% of men considering four or fewer treatment/medical aspects of prostate cancer. Men most commonly considered cancer stage (59%), urinary incontinence (55%) and impotence (51%) after surgery, and low overall mortality (45%). Uncertainty about probabilities for cure was reported by 43% of men and fear of cancer spread by 37%. Men also described uncertainty about the probabilities of side-effects (27%), decisional uncertainty (25%) and anticipated decisional regret (18%). Overall, 73% of men sought information about prostate cancer from external sources, most commonly the Internet, followed by family and friends. CONCLUSIONS: In general, men did not use information about medical treatments comprehensively or systematically when making treatment decisions, and their processing of medical information was biased by their previous beliefs about cancer and health. These findings have implications for the provision of informational and decisional support to men considering prostate cancer treatment. PMID- 11856107 TI - Prevalence of undiagnosed prostate cancer in men with erectile dysfunction. AB - OBJECTIVE: To explore the prevalence of prostate cancer in men presenting with erectile dysfunction (ED). PATIENTS AND METHODS: In a prospective study, 127 men with ED of at least 6 months duration underwent screening for prostate cancer using prostate specific antigen (PSA) and a digital rectal examination (DRE). Men with a high PSA level (> 4 ng/mL) had sextant biopsies taken under sedoanalgesia. The serum testosterone level was measured in all the men. RESULTS: Twenty-six men were aged < 50 years and all had a normal PSA level; of 101 men aged > 50 years, 20 had an abnormal PSA. The detection rate for prostate cancer using PSA and DRE was 5%, which was not significantly higher than in the general population. All the detected cancers were clinically significant (> T2a, Gleason grade > 4). Two of the five men diagnosed with prostate cancer were Afro-Caribbean. Of the 127 men, 31% had a low serum testosterone level, but there was no association between testosterone and PSA levels. CONCLUSIONS: Prostate cancer is no more common in men with ED than in the normal male population. Therefore, routine screening for prostate cancer in men with ED is not indicated. PMID- 11856108 TI - Antegrade scrotal sclerotherapy in the treatment of varicocele: a prospective study. AB - OBJECTIVE: To assess the results of our experience in correcting primary varicocele using a modified technique of antegrade scrotal sclerotherapy. PATIENTS AND METHODS: From December 1997 to February 2000, 201 patients with primary varicocele underwent antegrade scrotal sclerotherapy. Before treatment all patients were evaluated by a physical examination, colour Doppler ultrasonography of the spermatic cord, and abdominal and scrotal ultrasonography. Sperm samples were analysed only in patients aged > 18 years. The treatment was administered using a special venous catheter system with a Y-adapter. Complications after treatment and the rate of persistence of reflux were assessed 3 and 6 months after the antegrade scrotal treatment, by clinical examination and colour-Doppler ultrasonography of the spermatic cord. RESULTS: The mean operative duration was 15 min; there were no significant complications during treatment and all patients were discharged within 4 h. Complications after treatment occurred in 10 of the 201 patients (5%); four patients had epididymo-orchitis, three a scrotal haematoma, one a surgical wound infection, one persistent scrotal pain and one persistent flank pain on the same side as the surgery. Reflux was persistent in 12 patients (6%). CONCLUSIONS: Antegrade scrotal sclerotherapy is a simple, minimally invasive and highly effective technique for blocking spermatic vein reflux, with a low rate of complications. The technical changes simplified the cannulation of the spermatic vein and facilitated antegrade phlebography and sclerotherapy using the air-block technique. PMID- 11856109 TI - Laparoscopic Palomo varicocelectomy in the adolescent is safe after previous ipsilateral inguinal surgery. AB - OBJECTIVE: To evaluate the outcome of laparoscopic Palomo varicocelectomy (LPV) in young boys who had undergone previous ipsilateral inguinal surgery (in whom potentially the arterial supply to the testicles may be compromised) in an attempt to assess its safety for the collateral vascular supply in such cases. PATIENTS AND METHODS: Over a 5-year period (1995-2000) 44 patients underwent LPV, where both the spermatic artery and vein were ligated high above the internal ring. Thirteen patients had undergone previous ipsilateral inguinal surgery, which included inguinal hernia repairs in five, orchidopexy in two, communicating hydrocele repair in three and previous varicocele repair in three. All patients were followed clinically at 3 months and 1 year after surgery. RESULTS: There were no complications related to laparoscopy or varicocele ligation. No patient developed ipsilateral testicular atrophy; moreover the testis size remained stable or was associated with compensatory growth in all patients. CONCLUSION: Previous inguinal surgery involving the ipsilateral testicle does not appear to affect the collateral blood circulation to the affected testis in boys who undergo LPV mass ligation of the internal spermatic vein and artery. LPV for varicocele is safe in boys who have undergone previous inguinal surgery, suggesting that an adequate collateral blood supply is present. PMID- 11856110 TI - Urological complications of coitus. AB - OBJECTIVE: To ascertain the urological complications of coitus, as the proximity of the lower urinary tract to the organs of coitus exposes the tract to coital trauma. MATERIALS AND METHODS: Medline was searched from 1966 to 2000 to identify reports on coital injuries. Publications and relevant references were retrieved. Those reporting urological complications were selected for analysis. RESULTS: In all, 1454 cases of reported coital injuries were reviewed; 790 occurred in men while 664 occurred in women, mainly in the genital area. Physical urological complications were more common in men than in women. The injuries were often sustained during voluntary coitus, but one penile fracture was sustained during an attempted rape. The presentations included penile swellings and deviations, haemorrhage, erectile dysfunction and urinary incontinence. Complications included vesicovaginal fistulae, bladder and cavernosal ruptures, and urinary tract infections. Rare complications included isolated rupture of the penile vasculature. Major risk factors included penovaginal disproportion, excessive force at coitus, urethral coitus, fellatio and anal intercourse. Urethral injuries were the commonest complications; in men these were associated with 10 38% of penile fractures. The treatments included cold compress and anti inflammatory agents in contusions, repairs of lacerations, closure of fistulae and urethral and vaginal reconstruction. The results of treatment were essentially good. Recurrent penile fractures were reported. CONCLUSION: Coitus, although pleasurable, may be risky. The complications have been termed 'faux pas' implying that they are preventable. While the ultimate prevention is abstinence, this is an unrealistic prescription. Therefore, efforts are necessary to identify risk factors to enable preventive strategies. PMID- 11856111 TI - alpha-Catenin expression pattern and DNA image-analysis cytometry have no additional value over primary histology in clinical stage I nonseminomatous testicular cancer. AB - OBJECTIVE: To determine whether the alpha-catenin expression pattern and DNA content have additional value over primary tumour histology, including information on vascular invasion and tunica albuginea invasion, in detecting occult metastasis in patients with clinical stage I nonseminomatous germ cell tumours of the testis (NSGCT). PATIENTS AND METHODS: Fifty consecutive patients with clinical stage I NSGCT underwent retroperitoneal lymphadenectomy (RPLND) between 1986 and 1992. The orchidectomy specimens were histopathologically reviewed and immunohistochemically stained with mouse monoclonal anti-alpha catenin antibody. The presence of an aberrant or negative staining in >10% of the malignant cells was defined as abnormal; in all other cases tumours were classified as normal. Furthermore, intact nuclei were isolated from 50 microm thick paraffin sections of the primary tumour, Feulgen stained, and analysed with an image-analysis system. RESULTS: Of the 50 patients, 14 had positive retroperitoneal nodes (stage IIa, 28%), one pathologically staged I patient developed a lung metastasis (stage IV) within 3 months of RPLND. Univariate analysis showed that the presence of embryonal cell carcinoma, vascular invasion and tunica albuginea invasion were predictive for occult metastases. In multivariate logistic regression analysis only vascular and tunica albuginea invasion were significant. All 11 patients with no embryonal cell carcinoma in the primary tumour were classified as having pathological stage I disease. Also, the tumours which were DNA-diploid (three) or DNA-polyploid (two) were pathologically stage I. In screening for occult metastases the DNA content and the alpha-catenin expression pattern had no additional value. CONCLUSION: Vascular and tunica albuginea invasion have prognostic value in identifying patients with clinical stage I NSGCT at high risk for occult retroperitoneal disease. In contrast, the absence of embryonal cell carcinoma could predict all patients at low risk for metastasis. The DNA-ploidy also identified patients at low risk. Other DNA-analyses and the alpha-catenin expression pattern provided no additional information. Further studies are recommended to identify patients who are at low or high risk for metastasis. PMID- 11856112 TI - Low-flow priapism: risk factors for erectile dysfunction. AB - OBJECTIVE: To evaluate our policy of managing priapism for the success rate of the treatments, potency afterward, complications, and the risk factors responsible for erectile dysfunction in these patients. PATIENTS AND METHODS: The study included 50 patients (mean age 37.1 years, range 22-66) with a diagnosis of priapism (1981-1999). Their records were reviewed; 35 patients were available for a long-term evaluation. Factors assessed were the duration of priapism, history of previous recurrent attacks, possible underlying causes (e.g. haematological disorders, medications or trauma), relation to sexual stimulation, pain, and any attempt at previous management. A complete blood screen and blood gases were assessed in corporal aspirates. Duplex ultrasonography was used in all impotent patients at their follow-up. Early and late complications were reviewed, and patients asked about their erectile function before priapism, and any recurrence. RESULTS: The median (range) duration of priapism was 48 (6-240) h; almost half the patients presented > 48 h after the onset of priapism. Sixteen patients (32%) reported a history of previous recurrent attacks, of whom seven had a history of previous treatments. The main cause of priapism was idiopathic or intracavernosal injection with papaverine. All patients were initially treated by corporal blood aspiration and injection with ephedrine; if this failed or if the priapism was prolonged (> 48 h) various shunts were used. The hospital stay was significantly shorter among patients with papaverine-induced or brief priapism. In the long term follow-up of 35 patients (mean 66.4 months, range 3-220) only 15 (43%) reported preserved erectile function, and this was more likely in patients with brief priapism (< 48 h). Eight patients (23%) reported subsequent recurrent attacks of priapism; all were managed successfully as they presented shortly after their onset. Penile fibrosis was detected in 20 patients (57%), and was significantly more common in those with prolonged priapism (> 48 h) or from causes other than papaverine. The 20 impotent men evaluated by Doppler ultrasonography had severe echo-dense penile fibrosis and high end-diastolic velocities suggesting veno-occlusive incompetence in all except two. In five men with shunts cavernosography showed extensive venous leakage irrespective of site of the shunt. MRI in five patients with penile fibrosis showed heterogeneous areas of low signal intensity, corresponding with haemosiderin deposition and fibrosis. On univariate analysis the final result of management (complete detumescence or not), the duration of priapism and the presence of penile fibrosis significantly influenced erectile function. On multivariate logistic regression only the first remained significant. CONCLUSIONS: Low-flow priapism for > 48 h, failure to maintain complete detumescence after management, and marked penile fibrosis during the follow-up are the most significant risk factors responsible for erectile dysfunction, with failure to achieve complete detumescence the most detrimental. PMID- 11856113 TI - Further experience with the tubularized-incised urethral plate technique for hypospadias repair. AB - OBJECTIVES: To report further experience of hypospadias repair using the tubularized-incised urethral plate (TIP) technique and to evaluate the role of postoperative neourethral dilatation as a method of preventing complications. PATIENTS AND METHODS: The study included 64 patients (aged 2-18 years) who underwent TIP repair of hypospadias; 47 required a primary and 17 a secondary repair. After removing the stent they were randomized into two groups; group 1 (38 patients) underwent regular neourethral dilatation for 12 weeks and group 2 (26 patients) did not. Patients were followed for a mean (range) of 28 (6-52) months. RESULTS: The functional and cosmetic results were excellent in all patients in group 1, except for slight meatal regression in one patient (3%). In group 2, 17 patients had excellent results; eight (31%) were re-operated upon to correct complications, six developed a fistula (four of which were associated with meatal stenosis) and two developed a neourethral stricture. CONCLUSIONS: TIP urethroplasty is a versatile technique that provides an excellent functional and cosmetic outcome. Regular urethral calibration after repair should be considered as an integral part of the technique, to prevent neourethral and/or meatal stenosis with subsequent fistula formation. PMID- 11856114 TI - Overcoming parental intolerance in childhood nocturnal enuresis: a survey of professional opinion. PMID- 11856115 TI - Muscle precursor cell autografting in a murine model of urethral sphincter injury. AB - OBJECTIVE: To determine whether muscle precursor cells (MPCs) harvested from limb skeletal muscle can enhance the regeneration process of the striated urethral sphincter after injury. MATERIAL AND METHODS: Striated urethral sphincters of male mice were injured by an injection of a myotoxic substance (notexin). In the experimental group, 2 days after injury, MPCs were enzymatically harvested from striated muscles of the lower limbs and labelled with PKH 26, then immediately re injected into the injured urethral sphincter of the same animal. In the control group, saline buffer was injected instead of MPCs. Animals were killed 7 days or 1 month after injury and the sphincters removed for histological study (the presence of PKH 26-labelled myofibres, measurement of myofibre diameter and total number of myofibres). RESULTS: MPC autografting accelerated sphincter muscle repair, as shown by a higher myofibre diameter (P = 0.03) and number (P = 0.01) in the experimental group than in the controls at 7 days. One month after their injection MPCs were still detectable in the regenerating sphincters and participated in the formation of new myofibres. CONCLUSION: This study provides the experimental basis for a new therapeutic approach to urethral sphincter insufficiency after surgical or obstetrical injury, based on MPC autografting. PMID- 11856116 TI - Thrombospondin-1, vascular endothelial growth factor expression and their relationship with p53 status in prostate cancer and benign prostatic hyperplasia. AB - OBJECTIVE: To evaluate the expression of thrombospondin-1 (TSP-1, a potent inhibitor of angiogenesis) and vascular endothelial growth factor (VEGF, an important angiogenic factor in solid tumours) in prostate cancer, and their relationship with p53 status. PATIENTS AND METHODS: Using immunohistochemistry, the expression of VEGF, TSP-1 and p53 was assessed in 82 archival tissue specimens from 23 patients with benign prostatic hyperplasia (BPH), 22 with localized prostate cancer and 37 with metastatic prostate cancer. Seven of the last group had received androgen deprivation therapy. The relationship between the expression of VEGF, TSP-1 and p53 status was also evaluated with tumour grade and stage in patients with prostate cancer. RESULTS: The seven patients receiving hormonal treatment were excluded from the analysis because androgen deprivation significantly increased TSP-1 and decreased VEGF expression (both P < 0.01). Immunohistochemical analysis showed significantly higher VEGF and significantly lower TSP-1 expression (both P < 0.01) in prostate cancer than in BPH tissues. There was also significantly higher VEGF and significantly lower TSP-1 expression (both P < 0.05) in tissues from metastatic than localized prostate cancer. There was no significant correlation between VEGF or TSP-1 expression and Gleason score, but a significant inverse correlation between TSP-1 and VEGF expression. There was a significant association between VEGF expression and p53 status (P < 0.05), but TSP-1 expression was not associated with p53 status. CONCLUSIONS: Angiogenic factors, including VEGF and TSP-1, might be important in the development and progression of prostate cancer. These changes seem to be influenced by p53 status. Identifying the angiogenic factors involved in prostate cancer might lead to the development of diagnostic or therapeutic strategies based on anti-angiogenesis. PMID- 11856117 TI - Genetic changes in stage pT2N0 prostate cancer studied by comparative genomic hybridization. AB - OBJECTIVE: To identify chromosomal regions important for progression in clinically organ-confined prostate cancer, as the genetic changes underlying the development and progression of prostate cancer are poorly understood. MATERIALS AND METHODS: Comparative genomic hybridization (CGH) was used to search for DNA sequence copy-number changes in a series of 50 primary organ-confined prostate adenocarcinomas (pT2N0) removed by radical prostatectomy. RESULTS: CGH analysis indicated that 23 (46%) of the primary prostate adenocarcinomas showed chromosome alterations. The percentage of tumours with losses (38%) was higher than with gains (28%). Losses of 13q (24%), 8p (18%), 6q (10%), 16q (8%), 18q (6%) and 5q (6%) and gains of 17q (12%), 20q (12%), 9q (10%), 17p (8%) and 8q (6%) were the most frequent alterations. Amplifications were found at 8q24-qter. Minimal overlapping regions of loss, indicative of the presence of tumour-suppressor genes, were mapped to 13q21.1-q21.3 and 8p21.2, and minimal overlapping regions of gain, indicative of the presence of oncogenes, were found at 9q34.4-qter, 17q25-qter and 20q13.3-qter. There was a significant association between Gleason score and losses and gains (P = 0.003), and an association between chromosomal imbalance and high histological grade (P = 0.008). CONCLUSION: These results suggest that losses or gains of DNA in these regions are important for prostate cancer progression, and document the spectrum of chromosomal alterations in stage pT2N0 of clinically organ-confined prostate cancer. PMID- 11856118 TI - The rendezvous procedure to cross complicated ureteric strictures. PMID- 11856119 TI - Ectopic renal tissue in the gubernaculum associated with undescended testis. PMID- 11856120 TI - Spontaneous pelvi-cholecystic fistula. PMID- 11856122 TI - Underpowered studies. PMID- 11856123 TI - General surgery training crisis in America. PMID- 11856124 TI - Litigation for pneumothorax as a complication of fine-needle aspiration of the breast. AB - BACKGROUND: Pneumothorax is a rare but recognized complication of diagnostic needle aspiration of the breast. Two recent court cases alleging negligence have reached opposing opinions on very similar findings. METHODS: A Medline literature search was carried out. RESULTS: The reported incidence of pneumothorax after diagnostic aspiration of the breast in seven series varied between three in 100 and one in 10 000, but the weight of evidence tended towards the latter rate. Two studies reported that the complication is more common in the hands of trainees. It is not always possible to maintain the aspirating needle parallel or tangential to the chest wall. Pleural puncture may be more common than is apparent, and is most common in the tail of the breast in a thin woman. CONCLUSION: It is important that breast clinicians are aware of the risk of pneumothorax but, provided proper care has been taken, this complication is not the result of a negligent act. PMID- 11856125 TI - Systematic review of dynamic graciloplasty in the treatment of faecal incontinence. AB - BACKGROUND: The aim of this systematic review was to compare the safety and efficacy of dynamic graciloplasty with colostomy for the treatment of faecal incontinence. METHODS: Two search strategies were devised to retrieve literature from the Medline, Current Contents, Embase and Cochrane Library databases up until November 1999. Inclusion of papers depended on a predetermined protocol, independent assessments by two reviewers and a final consensus decision. English language papers were selected. Acceptable study designs included randomized controlled trials, controlled clinical trials and case series. Forty papers met the inclusion criteria. They were tabulated and critically appraised in terms of methodology and design, outcomes, and the possible influence of bias, confounding and chance. RESULTS: No high-level evidence was available and there were no comparative studies. Mortality rates were around 2 per cent for both graciloplasty and colostomy. Morbidity rates reported for graciloplasty appear to be higher than those for colostomy. Dynamic graciloplasty was clearly effective at restoring continence in between 42 and 85 per cent of patients, whereas colostomy is, by its design, incapable of restoring continence. However, dynamic graciloplasty is associated with a significant risk of reoperation. CONCLUSION: While dynamic graciloplasty appears to be associated with a higher rate of complications than colostomy, it is clearly a superior intervention for restoring continence in some patients. It is recommended that a comparative, but non randomized, study be undertaken to evaluate the safety of dynamic graciloplasty in comparison to colostomy, and that the procedure should be performed only in centres where it is carried out routinely. PMID- 11856126 TI - Randomized clinical trial of Ligasure versus open haemorrhoidectomy. AB - BACKGROUND: Postoperative pain associated with open haemorrhoidectomy remains problematic. Haemorrhoidectomy performed using bloodless bipolar diathermy- Ligasure--may have advantages over conventional open haemorrhoidectomy in terms of operating time and postoperative pain. METHODS: Thirty-four patients were randomized to undergo Ligasure (18 patients) or diathermy (16) haemorrhoidectomy. The operating time, amount of pain and postoperative analgesic requirement, postoperative complications and overall patient satisfaction were documented. RESULTS: The median duration of operation was shorter in the Ligasure haemorrhoidectomy group (5.1 versus 9.2 min; P < 0.001). There was no statistically significant difference in the postoperative pain score, but the median analgesic requirement was lower in the Ligasure group (850 versus 1600 mg tramadol; P = 0.013). Patient satisfaction was similar in both groups. CONCLUSION: Ligasure haemorrhoidectomy is quick and bloodless and, although as painful as diathermy haemorrhoidectomy, is associated with a reduced analgesic requirement. PMID- 11856127 TI - Long-term results of bilateral thoracoscopic splanchnicectomy in patients with chronic pancreatitis. AB - BACKGROUND: The management of pain in patients with chronic pancreatitis is difficult. The aim of this prospective study was to evaluate the early and long term pain relief provided by bilateral thoracoscopic splanchnicectomy. METHODS: From August 1995 to August 1999, 44 patients with chronic pancreatitis underwent bilateral thoracoscopic splanchnicectomy. Data were collected prospectively. Thirty-six patients required opioids. Pain intensity was registered before operation and at regular intervals after surgery by means of a visual analogue scale (VAS). Use of analgesics (opioids; non-steroidal anti-inflammatory drugs and acetaminophen; no analgesics or aminocetophen) was noted before and after splanchnicectomy. Median follow-up was 36 (range 12-60) months. RESULTS: The procedure was technically successful in 40 patients. Thirty-six patients had no complications. Eleven of 24 patients who have been followed up for 24 months or more had a significantly reduced VAS score at 2 years (median (range) 8.5 (7-10) versus 2.5 (0-5); P < 0.01). The cumulative rate of pain relief was 46 per cent 48 months after splanchnicectomy. CONCLUSION: Bilateral thoracoscopic splanchnicectomy alleviated pain in patients with chronic pancreatitis. It was associated with a low morbidity rate and no deaths. Pain eventually recurred in approximately 50 per cent. PMID- 11856128 TI - Serum amyloid A is a better early predictor of severity than C-reactive protein in acute pancreatitis. AB - BACKGROUND: Serum amyloid A (SAA) is an early and sensitive marker of the extent of tissue trauma and inflammation. The aim of this study was to compare the early prognostic accuracy of SAA with that of serum C-reactive protein (CRP) in acute pancreatitis. METHODS: In a prospective multicentre trial, plasma SAA and CRP levels were measured in patients with severe and mild acute pancreatitis, and in a control group with acute abdominal pain. Plasma samples were collected on admission and at 6-h intervals for 48 h, every 12 h between 48 and 72 h, then daily for 5 days. Plasma SAA was measured by a new enzyme-linked immunosorbent assay and CRP was measured by immunoturbidometry. RESULTS: There were 137 patients with mild and 35 with severe acute pancreatitis, and 74 control patients. SAA levels were significantly higher in patients with severe acute pancreatitis than in those with mild acute pancreatitis, on admission, at 24 h or less after symptom onset, and subsequently. Whereas plasma CRP concentration was also significantly higher in patients with severe acute pancreatitis on admission, it failed to distinguish mild from severe acute pancreatitis until 30 36 h after symptom onset. SAA levels predicted severity (sensitivity 67 per cent, specificity 70 per cent, negative predictive value 89 per cent, mean(s.d.) area under curve 0.7(0.05)) significantly better than CRP (57 per cent, 60 per cent, 84 per cent, 0.59(0.06) respectively) on admission (P = 0.02) and at 24 h following symptom onset (area under curve 0.65(0.09) versus 0.58(0.09) respectively; P < or = 0.02). CONCLUSION: Plasma SAA concentration is an early marker of severity in acute pancreatitis and is superior to CRP estimation on hospital admission and at 24 h or less after symptom onset. This study suggests that plasma SAA concentration is clinically useful, with the potential to replace CRP in the management of acute pancreatitis. PMID- 11856129 TI - Role of mast cells in the development of pancreatitis-induced multiple organ dysfunction. AB - BACKGROUND: Activated mast cells can produce and release a number of inflammatory mediators involved in the pathophysiology of acute conditions. The aim of the present study was to evaluate the role of activated tissue mast cells in the pathogenesis of multiple organ dysfunction syndrome following acute pancreatitis (AP). METHODS: AP was induced by the intraductal infusion of 5 per cent sodium taurodeoxycholate in the rat. Some 30 min before induction of AP, a mast cell stabilizer (sodium cromoglycate (SCG)) or antihistamines (pyrilamine, cyproheptadine, meclizine and amitriptyline) were administered intra peritoneally. Plasma exudation of radiolabelled albumin, histamine, myeloperoxidase (MPO), monocyte chemoattractant protein (MCP) 1 and adhesion molecules (platelet endothelial cell adhesion molecule (PECAM) 1 and L-selectin) were measured. RESULTS: The mast cell stabilizer significantly reduced plasma exudation in the pancreas, colon and lungs (P < 0.05), decreased the release of histamine at 1 h (P < 0.05), and reduced MPO activity and MCP-1 levels in the colon and lungs (P < 0.05) but not in the pancreas. Expression of PECAM-1 and L selectin on total circulating leucocytes in rats with AP and SCG pretreatment did not differ from that in sham controls, while levels in animals that had AP and saline pretreatment were half of those seen following sham operation. CONCLUSION: Activation of mast cells after induction of AP is involved in the development of endothelial barrier dysfunction in both the pancreas and extrapancreatic organs/tissues, particularly in the lungs and colon. This may, at least partly, contribute to the sequential development of multiple organ dysfunction and organ/tissue-specific endothelial barrier dysfunction. PMID- 11856130 TI - Extensive surgery for carcinoma of the gallbladder. AB - BACKGROUND: The purpose of this study was to clarify the efficacy of, and define the indications for, extensive surgery for gallbladder carcinoma. METHODS: Between 1979 and 1994, 116 patients with gallbladder carcinoma underwent operation. Radical resection was performed in 80 patients. RESULTS: In 68 patients with stage III or IV disease, extensive resection including extended right hepatectomy (n = 40), pancreaticoduodenectomy (n = 23) and/or portal vein resection (n = 23) was employed to achieve complete tumour excision. The hospital mortality rate was 18 per cent. The postoperative 3- and 5-year survival rates were 44 and 33 per cent respectively in the patients with stage III disease (n = 9), and 24 and 17 per cent respectively in patients with stage IV (M0) disease (n = 29). In contrast, the postoperative survival rate for the 30 patients with stage IV (M1) disease (7 per cent at 3 years and 3 per cent at 5 years) was worse than that for patients with stage III and stage IV (M0) disease (P = 0.009 and P = 0.062 respectively). CONCLUSION: Radical resection should be undertaken for stage III and stage IV (M0) gallbladder cancer. Although portal vein resection and/or pancreaticoduodenectomy did not contribute to long-term survival, better survival was obtained than that for the unresected patients. PMID- 11856131 TI - Effects of experimental lower-limb ischaemia-reperfusion injury on the mesenteric microcirculation. AB - BACKGROUND: Ischaemia-reperfusion (I-R) of the leg is associated with functional and structural changes in the intestine. This study assessed whether acute hind limb I-R in rats induced a reduction in perfusion and/or signs of an inflammatory response in the intestine. METHODS: Rats were subjected to 2 h of unilateral hind limb ischaemia followed by 2 h of reperfusion (I-R group, n = 9) or to a sham procedure (control group, n = 9). Mesenteric microvascular diameters, red blood cell velocity, blood flow and leucocyte-vessel wall interactions during reperfusion were measured using intravital microscopy. RESULTS: Blood pressure and heart rate decreased from 30 min of reperfusion onwards in the I-R group compared with controls. From 15 min after the start of reperfusion, mesenteric arteriolar and venular red blood cell velocity and blood flow decreased by 40-50 per cent. Microvascular diameters and leucocyte-vessel wall interactions did not change. CONCLUSION: Restoration of blood flow to an acutely ischaemic hind limb led to a significant decline in the splanchnic microcirculatory blood flow. There were, however, no signs of an early inflammatory response in the gut. PMID- 11856132 TI - Comparison of colour Doppler ultrasonography, ascending phlebography and clinical examination in the diagnosis of incompetent calf perforating veins. PMID- 11856133 TI - Outcome after septic complications in J pouch procedures. AB - BACKGROUND: Despite the wide range of surgical approaches for pouch salvage, septic complications are among the main causes of pouch failure. METHODS: This study analysed the mode and outcome of various therapeutic approaches for pouch salvage and the impact of time of diagnosis, localization and form of septic complications on the risk of pouch failure in 131 patients with septic complications of a total of 706 patients undergoing a J pouch procedure. RESULTS: Septic complications consisted of 76.3 per cent fistulas, 15.3 per cent anastomotic breakdowns and 8.4 per cent pelvic abscesses. A total of 107 patients (81.7 per cent) with septic complications required a mean of 2.2 surgical procedures. The frequency of permanent defunctioning and excision of the pouch in the 131 patients with septic complications was 23.7 and 6.1 per cent respectively. The estimated cumulative 3-, 5- and 10-year rate of pouch failure in patients with septic complications was 19.6, 31.1 and 39.2 per cent respectively. The risk of pouch failure was significantly affected by the site of origin of septic complications (P = 0.02). The 5-year pouch failure rate increased in a subgroup of patients with septic complications at the pouch-anal anastomosis when the anal sphincter was involved (50.1 versus 29.2 per cent; P = 0.18). CONCLUSION: Pouch failure as a result of septic complications may occur several years after ileal pouch-anal anastomosis. For prevention of pouch failure, surgery for septic complications is required in a high percentage of patients and repeated attempts are justified. Follow-up studies are required for further analysis of pouch failure. PMID- 11856134 TI - Evidence for separate disease phenotypes in intestinal Crohn's disease. AB - BACKGROUND: This study investigated the hypothesis that separate phenotypes of Crohn's disease exist which display differing patterns of recurrence with a tendency to preservation of phenotype between serial operations. METHODS: Some 483 abdominal operations (278 patients) were identified from a prospectively compiled database. Patterns of recurrence (reoperation) were analysed by Kaplan Meier plots and log rank tests according to disease phenotype (perforated, stenosed or ulcerated). Serial operations were analysed by agreement of phenotype and microscopic features of disease using kappa statistics and correlation coefficients. RESULTS: There was no significant difference in recurrence according to disease phenotype (median reoperation-free survival time 43.0, 50.2 and 47.9 months for perforated, stenosed and ulcerated types respectively; log rank chi(2) = 3.5, P = 0.18). There was poor agreement in phenotype between serial operations (kappa = 0.22 for first/second operation and kappa= 0.15 for second/third operation) and no significant correlation between pathological features was identified (r between -0.19 and 0.48). CONCLUSION: No evidence was found for the existence of separate disease phenotypes with differing natural histories or underlying pathological characteristics. PMID- 11856135 TI - Cancer recurrence along the pouch longitudinal suture line after colonic J pouch anal anastomosis. PMID- 11856136 TI - Cytoreduction including total gastrectomy for pseudomyxoma peritonei. AB - BACKGROUND: Cytoreductive surgery supplemented by perioperative intraperitoneal chemotherapy is a therapeutic option for selected patients with pseudomyxoma peritonei syndrome. In some patients, the stomach and/or its vascular supply are so covered by mucinous tumour that total gastrectomy is required for complete resection. METHODS: Forty-five patients underwent total gastrectomy with a temporary diverting jejunostomy as part of the surgical treatment of pseudomyxoma peritonei syndrome of appendiceal origin. Heated intraoperative intraperitoneal chemotherapy with mitomycin was used in all patients, and 36 had early postoperative intraperitoneal 5-fluorouracil. To date, 39 patients have had second-look surgery and stoma closure; 37 had additional perioperative intraperitoneal chemotherapy. A prospective database was maintained on all patients. RESULTS: The median age was 47 (range 33-66) years. Median interval from diagnosis of pseudomyxoma peritonei to definitive cytoreductive surgery was 23 (range 0-140) months. Six patients presented with intestinal obstruction. The need for gastrectomy was predicted before operation by abdominal computed tomography. Mean operative time was 13 (range 9-17) h. Mean intraoperative requirement for packed red blood cells was 3.0 units, and that for fresh frozen plasma was 9.9 units. Six peritonectomy procedures, including total gastrectomy, were required for complete cytoreduction. All except seven patients were maintained on parenteral nutrition before second-look surgery for jejunostomy closure. All but two patients have resumed oral nutrition with discontinuation of parenteral feeding. There was one postoperative death and one late death. Thirty seven patients are alive and disease-free, 0-56 months after initiation of treatment. CONCLUSION: Total gastrectomy with a temporary diverting jejunostomy may be used to facilitate complete cytoreduction in patients with advanced pseudomyxoma peritonei syndrome. PMID- 11856137 TI - Significance of immunohistochemical nodal micrometastasis as a prognostic indicator in potentially curable oesophageal carcinoma. AB - BACKGROUND: The number of positive lymph nodes is an important prognostic predictor in patients with oesophageal cancer. However, the significance of nodal micrometastasis in patients with overt nodal metastasis is unknown. The aim of this study was to clarify the clinical implications of nodal micrometastasis in patients undergoing curative oesophagectomy for oesophageal cancer. METHODS: Cervical, mediastinal and abdominal lymph nodes systematically removed from 104 patients with oesophageal cancer were examined immunohistochemically to detect cells that stained positively for cytokeratins with the monoclonal antibody cocktail AE1/AE3. The postoperative course and survival rates were compared among patients with and without micrometastases, after numerical classification of overt metastatic nodes (none, between one and four, five or more). RESULTS: Univariate analysis showed T stage, nodal micrometastasis and number of overt nodal metastases to be significant prognostic factors after oesophagectomy. Multivariate analysis revealed nodal micrometastasis and number of overt nodal metastases to be independent prognostic factors. The presence of micrometastases had a significant adverse effect on postoperative survival in patients with no overt metastasis and in patients with one to four overt metastatic nodes, but no such impact in patients with five or more overt metastatic nodes. CONCLUSION: Assessment of nodal status by both histological examination for overt metastases and immunohistochemical examination for micrometastases is useful in stratifying patients undergoing curative oesophagectomy. PMID- 11856138 TI - Length of the antral segment in pylorus-preserving gastrectomy. AB - BACKGROUND: Pylorus-preserving gastrectomy (PPG) has the advantages of preventing dumping syndrome and duodenogastric reflux. However, time was required to reduce symptoms of gastric fullness or to improve food intake after surgery. The aim of this study was to investigate whether the length of the retained antral segment influenced the postoperative symptoms in patients who underwent PPG. METHODS: Thirty patients with early gastric cancer who underwent PPG were divided into two groups according to the length of the retained antrum. In group 1 (n = 20) the transection was 1.5 cm and in group 2 (n = 10) it was 2.5 cm proximal to the oral edge of the pyloric ring. The subjects were interviewed and examined periodically to assess symptoms, food intake, body-weight and serum nutritional variables. Endoscopy and a radioisotope gastric emptying test were performed 1 year after the operation. RESULTS: In group 1 gastric fullness after meals was common, resulting in poor food intake and poor recovery of body-weight. A high incidence of food residue in the remnant stomach was found in group 1. With regard to gastric emptying, seven of 20 patients in group 1 and one of ten in group 2 had a delayed emptying curve 1 year after operation. CONCLUSION: Group 2 was superior to group 1 in terms of postprandial symptoms, food intake, recovery of body weight and gastric emptying. The length of the retained antrum may play an important role in the motility of the pyloric ring following PPG. PMID- 11856139 TI - Long-term results of antireflux surgery indicate the need for a randomized clinical trial. AB - BACKGROUND: Well conducted, comparative trials of laparoscopic versus open antireflux surgery with an adequate patient enrollment are few and they do not demonstrate obvious advantages for the laparoscopic approach except for a marginal gain in shorter hospital stay. The aim of this study was to compare the effectiveness of laparoscopic and open procedures. METHODS: Two unselected groups of 230 patients were identified through a register of all inpatient public care in Sweden. Outcomes of laparoscopic and open antireflux surgery were compared using a disease-specific questionnaire 4 years after operation. RESULTS: Failure and dissatisfaction were significantly more common in the laparoscopy group than among patients having conventional open surgery. Treatment failure rates were 29.0 and 14.6 per cent respectively (P = 0.004). Dissatisfaction rates were 15.0 and 7.0 per cent respectively (P = 0.005). There was no other questionnaire item for which the proportion of failures differed significantly between the two groups. CONCLUSION: This study does not support the presumption that laparoscopic antireflux surgery is to be preferred to the open procedure. It is strongly recommended that a randomized controlled trial be conducted. PMID- 11856140 TI - Endoscopic ultrasonography in the diagnosis and management of suspected upper gastrointestinal submucosal tumours. AB - BACKGROUND: 'Submucosal' lesions in the upper gastrointestinal tract are often difficult to evaluate. Endoscopic ultrasonography (EUS) provides high-quality information about the nature of these lesions and may assist management. This study assessed the use of EUS in the evaluation and management of upper gastrointestinal submucosal tumours. METHODS: Forty-four consecutive patients were referred with suspected upper gastrointestinal submucosal lesions for EUS. All examinations were performed by one of two experienced endosonographers. RESULTS: Most patients were referred for EUS with a suspected gastrointestinal stromal tumour. The pre-EUS diagnosis did not correlate with the EUS diagnosis in 25 of 44 patients. Pathological correlation was possible in 16 patients, and the EUS diagnosis was confirmed in each case. Based on the findings at EUS, 12 patients underwent resection. Pathology confirmed the ultrasonographic findings in these patients. Follow-up EUS to monitor lesions was recommended in a further three patients. CONCLUSION: EUS is safe and provides useful information in many patients with suspected submucosal tumours. EUS may reveal unsuspected findings and can increase physician certainty, allowing accurate diagnosis and facilitation of the management process. PMID- 11856141 TI - Abnormal expression of E-cadherin and beta-catenin may be a molecular marker of submucosal invasion and lymph node metastasis in early gastric cancer. AB - BACKGROUND: Impaired expression of E-cadherin and alpha- and beta-catenin is frequently observed in several human cancers. The aim of this study was to examine immunohistochemical expression of these adhesion molecules, focusing on early gastric carcinomas, and to investigate differences between differentiated and undifferentiated gastric cancer at the early phase of carcinogenesis. METHODS: Immunohistochemical staining of E-cadherin and alpha- and beta-catenin was performed using specimens from 143 patients with early gastric cancer. RESULTS: Abnormal E-cadherin and beta-catenin staining correlated with depth of tumour invasion in differentiated-type tumours. In contrast, abnormal staining was frequently found even in intramucosal carcinoma of undifferentiated-type tumours, suggesting an apparent difference in the onset of E-cadherin-catenin complex abnormality between the two cancer types. Absent staining of beta-catenin was associated with lymph node metastasis. Multivariate analysis revealed abnormal E-cadherin expression as an independent factor that correlated with submucosal invasion in early gastric cancer. CONCLUSION: Abnormal E-cadherin expression is a possible marker of submucosal invasion in differentiated-type early gastric cancer and absent beta-catenin staining could be used as a predictor of lymph node metastasis in both types. PMID- 11856154 TI - Fluorouracil, doxorubicin, and cyclophosphamide followed by tamoxifen as adjuvant treatment for patients with stage IV breast cancer with no evidence of disease. AB - We conducted a single-institution study to determine whether local therapy plus six cycles of chemotherapy with 5-fluorouracil, doxorubicin, and cyclophosphamide (FAC) followed by 5 years of tamoxifen is superior to local treatment alone in terms of disease-free survival (DFS) and overall survival (OS) in patients with stage IV breast cancer with no evidence of disease (stage IV-NED breast cancer). Patients with breast cancer were eligible if they had histologic proof of a locoregional or distant recurrence that had been curatively resected, irradiated, or both and had no other evidence of disease. Patients who had received prior anthracycline therapy were not eligible. All patients received six cycles of intravenous FAC, with cycles repeated every 3 weeks. After completion of chemotherapy, patients whose tumors had not previously demonstrated resistance to tamoxifen and had positive or unknown estrogen receptor status received tamoxifen 20 mg by mouth daily for 5 years. Patients in this study were compared with a historical control population (patients with stage IV-NED breast cancer who never received systemic therapy) as well as with the patients in two previously reported trials of chemotherapy for stage IV-NED disease. Forty-seven patients were registered, but only 45 were evaluable. There was a highly statistically significant difference ( p < 0.001) in OS and DFS among the four groups, with patients in our most recent study having the best OS and DFS at 3 years compared with the control group (84% vs. 55% and 66% vs. 11%, respectively). When patients in all four groups were analyzed together in search of prognostic factors, we found that patients whose primary tumors had negative axillary lymph nodes had a statistically significant improvement in OS and DFS ( p < 0.01) compared with patients with positive axillary lymph nodes. No survival differences were found between patients with positive and those with negative hormone receptor status. This study demonstrates a benefit in terms of OS and DFS for patients with stage IV-NED breast cancer who receive doxorubicin-based adjuvant chemotherapy. The benefit was greater on patients with node-negative primary tumors. In patients with stage IV-NED disease, doxorubicin-based chemotherapy should be considered standard treatment after adequate local control is achieved. PMID- 11856155 TI - Optimal sentinel node examination and a new strategy for axillary control in breast cancer. AB - The sentinel lymph nodes (SLNs) have been reported to reflect the other nodal status in breast cancer, and it is thought that axillary dissection can be avoided in SLN-negative patients, whereas an additional axillary dissection must be performed for SLN-positive patients. To avoid any unnecessary dissection, non SLN-negative patients should be distinguished among SLN-positive patients. In this study, an optimal histological examination of SLNs was investigated for a precise indication of additional axillary clearance. A modified radical mastectomy with an axillary dissection was performed for 61 patients. Technetium labeled tin colloids were used to identify the SLNs. In all lymph nodes that were diagnosed as negative at one section by hematoxylin and eosin (H&E) examination, another additional slice was produced for each lymph node. We evaluated how minute areas of cancer in the SLNs can indicate the non-SLNs to be negative. Among 1,092 lymph nodes previously diagnosed by H&E examination as negative at one section, 3 nodes were revealed to be positive in 46 SLNs (6.52%), whereas in additional slices, only 3 nodes out of 1,046 non-SLNs were positive (0.29%). The mean ratio of the cancer cell area to the whole area of SLN on the slice was 15.6% (10-18%) in 12 non-SLN-negative patients and 45.6% (30-83%) in 16 non-SLN positive patients. It is possible that additional axillary clearance is not needed for patients with a cancer area of less than approximately 20% in the SLN slice, based on an H&E staining evaluation of two slices. SLNs identified with radiolabeled tin colloids could reflect the other nodal status in breast cancer. There is a possibility that additional axillary clearance is not needed for patients with a cancer area of less than approximately 20% in the SLN slice. PMID- 11856156 TI - Competing considerations in regional nodal treatment for early breast cancer. AB - The purpose of this article was to review the patterns and morbidity of regional recurrence (RR) in patients with early breast cancer, efficacy of salvage therapy for RR, and complications of regional nodal treatment. A retrospective evaluation of 1,158 patients with stage I or stage II breast cancer treated with conservative surgery and radiotherapy (RT) between 1979 and 1994 was performed. Seven hundred fifty patients underwent axillary surgery, and 229 patients received RT as their only treatment of the regional lymphatics. Regional nodal RT was given to 168 patients who also had axillary surgery. The regional lymphatics of 11 patients were not treated. The patterns and morbidity of RR, relapse management, and complications related to regional nodal treatment were reviewed from the patients' records. With a median follow-up of 88 months, a total of 31 patients (2.7%) developed a RR. Nine of 31 patients (29%) with an RR experienced significant morbidity, including pain, fungating tumor, dysphagia, dyspnoea, and/or sensory motor changes at diagnosis. Nineteen patients (61%) had symptomatic residual or progressive regional disease after salvage therapy at last follow-up or death. Six of nine patients (67%) who developed an isolated axillary recurrence and underwent salvage surgery had no further axillary recurrence. The addition of regional nodal RT to breast irradiation significantly increased the incidence of symptomatic pneumonitis (1% without regional nodal RT and 4% with regional nodal RT, p < 0.001). Combined axillary dissection and nodal irradiation resulted in a significantly higher incidence of arm edema compared with either alone (9.5% with axillary dissection, 6.1% with RT to the axilla and supraclavicular fossa, and 31% with combined modality therapy, p < 0.001). Five of 380 patients (1%) who received RT to the axilla and/or supraclavicular fossa developed a transient brachial plexus neuropathy. Although RR was uncommon in patients treated with axillary surgery and/or regional nodal irradiation, salvage therapy failed to eradicate the recurrence in approximately two thirds of the patients with a RR. Ongoing research is essential to optimize regional control with an acceptable level of risk of treatment complications. Sentinel lymph node biopsy, if validated as an accurate method of staging the axilla in patients with breast cancer, would allow selective avoidance of regional nodal treatment and hence the associated morbidity. PMID- 11856157 TI - Limitations of the Gail model in the specialized breast cancer risk assessment clinic. AB - The Gail model is a risk assessment tool that is accurate for general breast cancer risk screening. Because of the limited way that this model incorporates family history information, however, there are concerns that it may underestimate risk for many women attending specialized breast cancer risk assessment clinics. We collected comprehensive breast cancer risk factor information for 213 women attending a specialized breast cancer risk assessment clinic using a modified version of the CancerGene software. Breast cancer risk was calculated using the models of Gail and Claus as well as BRCAPRO and the tables of Bodian (for women with lobular neoplasia). Eighty-six percent of the women had a family history of breast cancer. Although 74% of women had risk factor histories that are thought to confound the Gail model (family history of breast cancer in second-degree relatives, family history of breast cancer before the age of 50, family history of bilateral breast cancer, family history of ovarian cancer, or personal history of lobular neoplasia), the inclusion of other models increased the risk level assignment in only 13% of the cases. We conclude that the Gail model is an appropriate risk assessment tool for most women attending specialized clinics, although the inclusion of models better able to account for family history information and personal history of lobular neoplasia is required to accommodate all women. PMID- 11856158 TI - Long-term follow-up of upper-body function among breast cancer survivors. AB - We enrolled a cohort of 303 stage I or stage II breast cancer patients diagnosed in Boston, MA, between October 1992 and December 1995. We followed the patients by interview and medical record abstract for 5 years (a) to characterize the incidence and predictors of upper-body function decline and (b) to characterize the incidence and predictors of recovery of upper-body function. The incidence of decline in the first year after therapy (17.7/100 person years) was substantially higher than in the subsequent 4 years of follow-up (11.0/100 person-years, p value for test of homogeneity equal 0.028). With only one exception, no patient characteristic, therapy component, or disease trait was associated with decline over the full follow-up period. Women with less than a high school education had an adjusted relative hazard of decline of 2.3 (95% CI, 1.4-3.7) compared with women with a high school education or more, possibly reflecting occupational or environmental insults that predispose to functional impairment. Women who had reported a decline in upper-body function and who subsequently saw their breast cancer specialist were 4.8-fold more likely to report that they had recovered their upper-body function at their next interview (95% CI, 2.0, 12). This finding suggests that attention to upper-body function during follow-up visits may facilitate recovery. PMID- 11856159 TI - Breast cyst recurrence after postaspiration injection of air. AB - Breast cysts are a common cause of breast pain, lumps, and patient anxiety. Older studies incidentally reported a decrease in cyst recurrence when air was injected for diagnostic purposes after aspiration. The purpose of this study was to determine the incidence of cyst recurrence after ultrasound-guided aspiration and injection of air for therapeutic purposes. In this study, we retrospectively reviewed 113 cysts aspirated in 90 women; 88 had air injected for therapeutic purposes after aspiration, and 25 did not. Subsequent mammograms and sonograms were reviewed (mean follow-up 21 months, range of 8-36 months) to assess for recurrence. Demographic and imaging features were not different between the two study groups. Of the 88 cysts that had air injected after aspiration, 14 recurred (16%) compared with 20 of 25 cysts (80%) that did not have air injected following aspiration. Cyst recurrence correlated with air injection, but did not correlate with cyst size, menopausal status, or use of hormone replacement therapy. Only one patient complained of pain, and no complications occurred. The injection of air into cysts following aspiration may be useful for therapeutic purposes. PMID- 11856160 TI - p53 and Ki-67 expression as prognostic factors in cystosarcoma phyllodes. AB - We have reviewed the histopathological, clinical outcome and immunohistochemical status in 21 women with cystosarcoma phyllodes (CSP) tumors of the breast. We assessed 12 tumors as histopathologically benign and 9 tumors as malignant. The median patient ages in benign and malignant CSP tumors were 39.6 and 45.4 years of age, respectively. The stromal cellularity, stromal cellular atypism, high mitotic activity, atypic mitoses, stromal overgrowth, infiltrative tumor contour, and heterologous stromal elements were significant features of the malignant CSP tumors. Benign CSP tumors were predominantly of fibroadenomatous architecture with cellular stroma (mild or moderate) and some distortion and elongation of glandular elements. Five malignant CSP tumors were stained positively with p53, and 6 malignant CSP tumors were stained immunohistochemically with Ki-67. All benign CSP tumors were negatively stained for p53 and Ki-67. The patients with benign CSP tumors were treated with local excision ( n=11) and with subcutaneous mastectomy ( n=1). Malignant CSP tumors were treated with wide local excision ( n=1), partial mastectomy ( n=1), simple mastectomy ( n=2), and modified radical mastectomy ( n=5). Two patients with a high mitotic rate and high values of p53 and Ki-67 received additional radiotherapy and chemotherapy. One case had liver metastasis. This tumor had high mitotic figures, stromal overgrowth, severe stromal cellularity, and 20% Ki-67 and mild p53 positivity. We suggest that p53 and Ki-67 can play an important role in predicting prognosis and yielding additional therapy besides conventional prognostic factors in the treatment of the CSP patients. PMID- 11856161 TI - "Swiss-roll" operation for giant fibroadenomas. AB - Giant fibroadenomas can produce breast deformity and should be excised. Several techniques have been discussed, including a sub-mammary incision, which often leave a disfiguring scar. We describe a new technique for removal of such large masses via a small cosmetic circumareolar incision. PMID- 11856162 TI - Cellular angiolipoma of the breast: immunohistochemical study and review of the literature. AB - We report a rare case of cellular angiolipoma of the breast in a 73-year-old woman with abnormal mammographic findings. Histopathologic examination of the tumor revealed cellular spindle cell areas associated with collapsed and patent blood vessels in a background of mature adipose tissue. A salient feature of this tumor, which was an important clue to the diagnosis, was the presence of intravascular fibrin thrombi. Immunohistochemical studies showed reactivities for endothelial and muscle markers, highlighting the obscured angiomatous component in the cellular areas of the tumor. Failure to recognize this entity may potentially lead to diagnostic pitfalls that include angiosarcoma and Kaposi's sarcoma, two of the better recognized entities in this region. PMID- 11856163 TI - Osseous metaplasia of the breast: diagnosis with stereotactic core biopsy. AB - There are approximately 200 reported cases of breast tumors containing areas of bone. The majority of the neoplasms are sarcomas, phylloides tumors, or fibroadenomata. We present a case of osseous metaplasia mammographically detected by clustered heterogeneous calcifications. Stereotactic core biopsy revealed the presence of well-formed bone tissue without associated neolplasia. The case represents the first reported case of mammographically detected osseous metaplasia confirmed by core biopsy. PMID- 11856164 TI - Adenoid cystic carcinoma of the breast: mammographic, sonographic, and pathological correlation. PMID- 11856165 TI - Aggressive fibromatosis of the breast: a case report and literature review. AB - Aggressive fibromatosis is a rare, locally aggressive disease. It constitutes 0.3% of all solid tumors, but the tumor is rarely seen in the breast, particularly without pectoral muscle and fascial involvement. The etiology is unknown, but an association with Gardner's syndrome has been described. Clinical and imaging findings simulate breast carcinoma. A case in a 53-year-old female patient is reported. PMID- 11856166 TI - Laughing with breast cancer. PMID- 11856167 TI - The main problem in treating nonpalpable breast lesion is the need for accurate localization during surgery. PMID- 11856168 TI - Bacterial interactions with the autophagic pathway. AB - Bacteria have evolved a variety of mechanisms to invade eukaryotic cells and survive intracellularly. Once inside, bacterial pathogens often modulate their phagosome to establish an intracellular niche for survival and replication. A subset of intracellular pathogens, including Brucella abortus, Legionella pneumophila and Porphyromonas gingivalis, are diverted from the endosomal pathway to the auto-phagic pathway. Once within the autophagosome, each in some way presumably modifies this compartment to establish an environment necessary for its survival. Transit into autophagosomes represents an avenue by which to escape host defences. In this review, we examine the biochemical and morphological evidence for the survival of some bacterial pathogens by replicating within an autophagosome-like compartment. PMID- 11856169 TI - Cell vacuolization induced by Helicobacter pylori VacA cytotoxin does not depend on late endosomal SNAREs. AB - Cellular vacuoles induced by the Helicobacter pylori vacuolating cytotoxin VacA originate from late endosomal compartments. Their biogenesis requires the activity of both rab7 GTPase and the ATPase proton pump. The toxin has been suggested to cause an increased luminal osmotic pressure via its anion-specific channel activity localized on late endosomal compartments after endocytosis. Here, we show that the extensive membrane fusion that takes place in the transition from the small late endosomal compartments to the large vacuoles does not depend on soluble N-ethylmaleimide-sensitive factor (NSF) attachment protein receptor (SNARE) proteins. The process of vacuolization leads to disappearance of the large array of internal membranes of late endosomes. We suggest that most of the vacuole-limiting membrane derives from internal membranes. PMID- 11856170 TI - Requirement of Rho-family GTPases in the invasion of Type 1-piliated uropathogenic Escherichia coli. AB - Bladder infections caused by uropathogenic Escherichia coli (UPEC) depends on the ability of E. coli to express type 1 pili. The adhesive component of the pilus, FimH, mediates the invasion of E. coli into the bladder epithelium, a mechanism that facilitates the survival and persistence of E. coli in the bladder. The invasion mechanism requires actin polymerization, focal adhesion kinase phosphorylation and PI 3-kinase activation as well as the formation of FAK/PI 3 kinase and downstream vinculin/alpha-actinin complexes. In this study, we report a role for Rho-GTPase family members, namely RhoA, Cdc42 and Rac1, in the invasion process. Internalization of type 1-piliated E. coli (fimH+) and FimH coated micro-spheres was inhibited by compactin, a pan-Rho-GTPase inhibitor and dominant negative isoforms of Rac1 and Cdc42. Expression of active Rac1 induced an internalization of E. coli that was insensitive to wortmannin and genistein. Expression of constitutively active Cdc42 induced the formation of FAK/PI 3 kinase and vinculin/alpha-actinin complexes whereas active Rac1 induced only a vinculin/alpha-actinin complex. Taken together, these data suggest that FimH mediated invasion is dependent on GTP-binding protein activity that involves Cdc42 and PI 3-kinase activation probably upstream of Rac1. PMID- 11856171 TI - Species-specific cell adhesion of enteropathogenic Escherichia coli is mediated by type IV bundle-forming pili. AB - Enteropathogenic Escherichia coli (EPEC) is a causative agent of diarrhoea in humans. Localized adherence of EPEC onto intestinal mucosa was reproduced in an in vitro adherence assay with cultured human epithelial cells. We found that the efficiency of EPEC adherence to a mouse-derived colonic epithelial cell line, CMT 93, was remarkably lower than its adherence to human-derived intestinal cell lines, such as Intestine-407 or Caco-2. Although EPEC did adhere to some cell lines derived from non-human species, fixing the cells with formalin to inactivate one or more formalin-sensitive factors allowed us to observe species specific differences in EPEC adherence. In contrast to these results, an EPEC mutant that is defective in bundle-forming pili (BFP) production adhered as efficiently to CMT-93 cells as to Caco-2 cells. Furthermore, Citrobacter rodentium expressing BFP adhered to Caco-2 cells much more efficiently than to CMT-93 cells. Finally, a purified BfpA-His6 fusion protein showed higher affinity for Caco-2 cells than for CMT-93 cells, and inhibited EPEC adherence. Following BFP-mediated adherence, secretion of EspB from adherent bacteria and reorganization of F-actin in the host cells was observed. EPEC adhering to CMT-93 cells induced far less secretion of EspB, or reorganization of F-actin in the host CMT-93 cells, than did EPEC adhering to Caco-2 cells. These results indicated that BFP plays an important role in the cell-type-dependent adherence of EPEC and in the progression to the later steps in EPEC adherence. PMID- 11856172 TI - The invasion-associated type III secretion system of Salmonella enterica serovar Typhimurium is necessary for intracellular proliferation and vacuole biogenesis in epithelial cells. AB - Type III secretion systems (TTSS) are used by Gram-negative pathogens to translocate proteins into eukaryotic host cells. Salmonella enterica serovar Typhimurium (S. Typhimurium) has two of these specialized systems, which are encoded on separate Salmonella pathogenicity islands (SPI-1 and SPI-2) and translocate unique sets of effectors. The specific roles of these systems in Salmonella pathogenesis remain undefined, although SPI-1 is required for bacterial invasion of epithelial cells and SPI-2 for survival/replication in phagocytic cells. However, because SPI-1 TTSS mutants are invasion-incompetent, the role of this TTSS in post-invasion processes has not been investigated. In this study, we have used two distinct methods to internalize a non-invasive SPI-1 TTSS mutant (invA) into cultured epithelial cells: (i) co-internalization with wild-type S. Typhimurium (SPI-1-dependent) and (ii) complementation with the Yersinia pseudotuberculosis invasin (inv) gene (SPI-1-independent). In both cases, internalized invA mutants were unable to replicate intracellularly, indicating that SPI-1 effectors are essential for this process and cannot be complemented by wild-type bacteria in the same cell. Analysis of the biogenesis of SCVs showed that vacuoles containing mutant bacteria displayed abnormal maturation that was dependent on the mechanism of entry. Manipulation of Salmonella-containing vacuole (SCV) biogenesis by pharmacologically perturbing membrane trafficking in the host cell increased intracellular replication of wild type but not mutant S. Typhimurium This demonstrates a previously unknown role for SPI-1 in vacuole biogenesis and intracellular survival in non-phagocytic cells. PMID- 11856173 TI - Delinking of S phase and cytokinesis in the protozoan parasite Entamoeba histolytica. AB - The alternation of DNA replication in S phase and chromosome segregation in M phase is a hallmark in the cell cycle of most well-studied eukaryotes and ensures that the progeny do not have more than the normal complement of genes and chromosomes. An exception to this rule has been described in cancer cells that occasionally become polyploid as a result of failure to restrain S phase despite the failure to undergo complete mitosis. Here, we describe the cell division cycle of the human pathogen, Entamoeba histolytica, which routinely accumulates polyploid cells. We have studied DNA synthesis in freshly subcultured cells and show that, unlike most eukaryotes, Entamoeba cells reduplicate their genome several times before cell division occurs. Furthermore, polyploidy may occur without nuclear division so that single nuclei may contain 1-10 times or more genome contents. Multinucleated cells may also accumulate several genome contents in each nuclei of one cell. Thus, checkpoints that normally prevent DNA reduplication until after cytokinesis in most eukaryotes are not observed in E. histolytica. PMID- 11856174 TI - Blast colony-forming cell binding from CML bone marrow, or blood, on stromal layers pretreated with G-CSF or SCF. AB - Blast colony-forming cells (CFU-BL) represent a specific subpopulation of special primitive progenitors characterized by colony formation only in close contact with a preformed stromal layer. CFU-BL derived from bone marrow of chronic myeloid leukaemia (CML) patients have been proved to adhere poorly to bone marrow derived stromal layers suggesting that the appearance of progenitors and precursors in the circulation is due to a defective adhesion of these cells to the bone marrow microenvironment. In the present experiments the effect of short term incubation of preformed normal bone marrow stroma on the adherence of CML derived CFU-BL was studied. For stroma cultures bone marrow cells were cultured in microplates in the presence of hydrocortisone. Cultures were used when stromal layers became confluent and no sign of haemopoiesis could be observed. CFU-BL were studied by panning plastic non-adherent mononuclear (PNAMNC) bone marrow or blood cells. 8.9 +/- 2.4 colonies/103 PNAMNC (six experiments) were formed from normal bone marrow on stromal layers and 4.8 +/- 2.1 colonies/103 PNAMNC (five experiments) from CML bone marrow. Colony formation from normal bone marrow was not increased if stromal layers were incubated with 100 ng/mL granulocyte colony stimulating factor (G-CSF) or stem cell factor (SCF). Incubation of stroma with G CSF or SCF, however, increased the colony formation of PNAMNC from CML bone marrow or blood significantly. These findings suggest that local concentration of haemopoietic growth factors at the time of panning may influence the attachment of CML progenitors to the stroma. PMID- 11856176 TI - Human glioma PKC-iota and PKC-betaII phosphorylate cyclin-dependent kinase activating kinase during the cell cycle. AB - Cell cycle phase transition is regulated in part by the trimeric enzyme, cyclin dependent kinase activating kinase (CAK) which phosphorylates and activates cyclin-dependent kinases (cdks). Protein kinase C (PKC) inhibitors prevent cell cycle phase transition, suggesting a fundamental role for PKCs in cell cycle regulation. We report that in glioma cells, CAK (cdk7) is constitutively associated with PKC-iota. In vitro phosphorylation, co-immunoprecipitation, and analysis of phosphorylated proteins by autoradiography indicate that CAK (cdk7) is a substrate for PKC-iota and PKC-betaII hyperphosphorylation. These results establish a role for PKC-iota and PKC-betaII in the activation of CAK during the glioma cell cycle. PMID- 11856175 TI - Extracellular matrix alters the relationship between tritiated thymidine incorporation and proliferation of MC3T3-E1 cells during osteogenesis in vitro. AB - Bone cells in vivo exist in direct contact with extracellular matrix, which regulates their basic biological processes including metabolism, development, growth and differentiation. Thus, the in vitro activity of cells cultured on tissue culture treated plastic could be different from the activity of cells cultured on their natural substrate. We selected MC3T3-E1 pre-osteoblastic cells to study the effect of extracellular matrix on cell proliferation because these cells undergo a progressive developmental sequence of proliferation and differentiation. MC3T3-E1 cells were cultured on plastic or plastic coated with ECM, fibronectin, collagen type I, BSA or poly l-lysine and their ability to proliferate was assessed by incorporation of [3H]dT or by enumeration of cells. Our results show that (1) ECM inhibits incorporation of [3H]dT by MC3T3-E1 cells; (2) collagen type I, but not BSA, poly l-lysine or fibronectin also inhibits incorporation of [3H]dT; (3) the level of ECM inhibition of [3H]dT incorporation is directly related to the number of cells cultured, but unrelated to the cell cycle distribution or endogenous thymidine content; (4) the kinetic profile of [3H]dT uptake suggest that ECM inhibits transport of [3H]dT from the extracellular medium, and (5) cell counts are similar in cultures whether cells are grown on plastic or ECM. These results suggest that decreased incorporation of [3H]dT by cells cultured on ECM is not reflective of bone cell proliferation. PMID- 11856177 TI - Influence of irradiation and pentoxifylline on histone H3 phosphorylation in human tumour cell lines. AB - Phosphorylation of histone H3 at Ser-10 correlates with chromatin condensation and this amino terminal modification is now recognized as a specific marker of mitosis. We have monitored the appearance of cells showing histone H3 phosphorylation in four human tumour cell lines to identify cell cycle progression after irradiation. In the human melanoma cell lines Be11 and MeWo and in the squamous cell carcinoma lines 4197 and 4451 a dose of 7 Gy of Co-gamma irradiation increases the number of cells binding anti-histone H3-P antibody 1-8 fold in a p53-independent manner. In the p53 mutant cell lines MeWo and 4451 H3-P phosphorylated cells can be detected as early as 30 min and show a maximum 1 h post-irradiation. In the cell lines Be11, 4197 and 4451 the early wave of H3 phosphorylated cells is followed by a second wave, which reaches a maximum 4.5-7 h post-irradiation and then declines. These events are attributed to damage induced cell cycle blocks in the G1 and G2 phase of the cell cycle. Addition of the dose modifying drug pentoxifylline before irradiation increases the appearance of cells showing early and the late H3 phosphorylation. When pentoxifylline is added 12-24 h post-irradiation when the cell cycle blocks have reached their maximum the appearance of cells with phosphorylated H3 increases 3 5-fold in the p53 mutant cell lines MeWo and 4451. These observations are consistent with the function of the drug as a G2 block abrogator. The large H3 phosphorylation signal in p53 mutant cells is consistent with early entry of a cohort of G2 cells into mitosis. The smaller H3-P signal in p53 wild type cells correlates with the lower proportion of stable G2 populations in G1 blocked cells. These results indicate that pentoxifylline influences the appearance of histone H3 phosphorylated cells in a manner strongly dependent on the number of cells in G2 phase. This suggests that addition of pentoxifylline indeed abrogates the G2 block and thereby facilitates early entry into mitosis. PMID- 11856178 TI - Estimation of directional division frequencies in vascular cambium and in marginal meristematic cells of plants. AB - Using simple arithmetical formulae, it is shown that, when the meristematic initial cells of a growing plant organ are arranged in a ring, the cellular dimensions predict the relative frequencies of anticlinal and periclinal divisions which these cells undergo. The pattern of cell file branching which appears during the course of development, and which is predicted by this mathematical model, is validated using data pertaining to the numbers and dimensions of initial cells within the secondary vascular cambium of hybrid aspen trees. Data pertaining to a second, simpler set of initial cells which comprises the outer cellular ring of the thallus of the alga Coleochaete orbicularis, and from which all the radial cell files of the circular disc-like thallus are descended, have also been used for model validation. Combining the mathematical approach to division frequencies with data of actual cell sizes permits inferences about the course of the increase of the number of cell files (generated by the anticlinal divisions) and the number of cells within each file (generated by the periclinal divisions) during the earlier stages of secondary tissue or thallus development, and also about how they will develop at future stages. The question whether or not cell division patterns conform to the geometry of the system in which the cells are embedded is also discussed. PMID- 11856181 TI - Sure Start: key principles and ethos. PMID- 11856182 TI - The school experience of the child with cancer. AB - OBJECTIVE: To review literature investigating school absence, behaviour problems and social relationships of children with cancer on return to school. Additionally, we reviewed interventions aimed at promoting successful school re entry for children with cancer, and increasing classmates and teachers understanding of childhood cancer. METHODS: Electronic searches were used to identify studies examining the effects of school re-entry on children with cancer. RESULTS AND CONCLUSIONS: School absences are higher for children with cancer than healthy children and those with other chronic conditions. Although there is mixed evidence about whether children have significant behavioural problems, studies involving social relationships generally conclude that children with cancer are more sensitive and isolated than peers, according to both peer and teacher report. PMID- 11856183 TI - Communicating about pupils in mainstream school with special health needs: the NHS perspective. AB - AIM: To add to previous research carried out with young people, parents and teachers, by investigating health staff's perspectives on the difficulties, and possibilities for, achieving good communication with school staff with regard to children in mainstream school with a chronic illness or physical disability. RESEARCH DESIGN AND METHODS: A qualitative research study was carried out in one NHS Trust. Twenty semi-structured interviews were carried out with a purposive sample of health staff to cover the spectrum of professionals who have responsibility for meeting the needs of children with a chronic illness or physical disability in mainstream schools (paediatricians, school doctors, school nurses, specialist nurses, health visitors, GPs, speech and language therapists, paediatric occupational therapists, paediatric physiotherapists, clinical psychologists and psychiatrists). RESULTS: The extent to which health professionals communicated with school staff, and the way in which they went about it, varied widely. Communication was facilitated by joint meetings, shared documentation, and local policy development. Sources of difficulty in communication between health and education staff were: the parent as a conduit of information; the practical difficulties of arranging meetings; and lack of knowledge about other professionals' roles. The ethos of the school with regard to health matters, and the flow of information within health services, also had an impact on the communication process. Participants' recommendations focused on two key issues: clarification of the roles of health and education staff with regard to this group of pupils; and how information should flow from health to school staff. DISCUSSION: Many of the findings parallel the previous research with teachers, indicating agreement between professionals from different agencies about aspects of the communication process which are problematic and require attention. The findings suggest that improving communication requires both joint work between health and education staff, and improvements to practice within each agency. PMID- 11856184 TI - Early rehabilitation service utilization patterns in young children with developmental delays. AB - OBJECTIVE: There is a growing demand for interdisciplinary rehabilitation services for children with developmental disabilities, however, information is lacking on service utilization patterns. This study describes the type, frequency and location of initial rehabilitation services provided to young children recently diagnosed with a developmental delay. METHODS: A telephone survey was conducted on a consecutive cohort of children referred to subspecialists for the first investigation of their delay. RESULTS: Out of 129 respondents, only 30 (23%) did not receive any rehabilitation services within the first 6 months following medical evaluation, and 61 (47%) received two or more services. Services were provided most frequently in the hospital setting, especially for occupational or physical therapy (73-80%). Services were less often rendered in a rehabilitation centre (5-13%) or in the community (< 10%). Services were provided privately for a subset, particularly for speech language pathology and psychology (34% and 30% respectively). Interventions were typically given weekly or bi weekly. Children receiving physical therapy were significantly younger whereas children referred to speech language pathology were older than the age of our cohort at intake. Overall, parents' educational level did not influence services received with the exception of private services, which were more likely to occur in children of fathers with university education. CONCLUSIONS: The findings would suggest that long waiting times and lack of resources may limit access to comprehensive services, particularly in community settings. Service utilization patterns were not consistent within types of developmental disability, suggesting that formal and co-ordinated interdisciplinary programmes are not in place for this population of interest. PMID- 11856185 TI - Citizens advice in primary care for families with young children. AB - BACKGROUND: Citizens advice can help clients avoid the worst consequences of poverty and hence has the potential to reduce inequalities in health. Previous studies of providing citizens advice from primary care have concentrated largely on older clients. AIM: To measure the benefits obtained by families with young children from a citizens advice service in primary care. METHOD: An advice worker was placed in three practices. Eligible families had a child less than 1 year of age. Quantitative analysis of data on benefits obtained by all families using the service was performed. Qualitative analysis of interviews with five users and five non-users of the service was undertaken. RESULTS: Out of 107 families recruited, 23 used the CAB service, seeking advice on 49 different problems and obtaining benefit on 32 of these, most frequently on welfare benefits, debt management and housing. One-off financial benefits totalled pound sterlings 17 857 with annual recurring payments of pound sterlings 6480. The interviews raised five general themes: (1) the stresses surrounding the birth of a child; (2) factors associated with uptake of the service; (3) the content and value of the advice received; (4) the benefits to their health of the advice; and (5) their perception of the links between health and financial adversity. CONCLUSIONS: Citizens advice offers considerable benefits to families with young children. It may have positive effects on maternal and child health. An advice service is a useful component of primary health care for young families. PMID- 11856186 TI - Growth of working boys in Jordan: a cross-sectional survey using non-working male siblings as comparisons. AB - OBJECTIVES: To study the effects of work on growth of Jordanian boys, aged 10-16 years, using non-working male siblings as controls. SETTING: The Jordanian areas of Irbid, Jarash and the North Jordan Valley. STUDY DESIGN: Cross-sectional survey of working boys and their non-working brothers. MAIN OUTCOMES: Height-for age z-score; weight-for-age z-score. METHODS: Working boys (103) and non-working male siblings (103) (nearest in age to the working child) were interviewed, with their mothers, in the family home. Heights and weights of the working boys and their non-working male siblings were measured, and capillary blood was taken for packed-cell volume estimation. Heights and weights were converted to z-scores, and means for all three outcomes were compared between working boys and non working siblings, using independent sample t-tests. The effect of the proportion of household income contributed by the working child's income on the main outcomes, among non-working siblings, was estimated by simple linear regression. RESULTS: Means for height-for-age z-score (p < 0.001), weight-for-age z-score (p < 0.001), and packed-cell volume (p < 0.001) among working boys were significantly lower than means for their non-working siblings. The main outcomes among non-working siblings were not significantly correlated with the proportion of household income contributed by the working child. CONCLUSIONS: The results of this study suggest that work among boys aged 10-16 years in Jordan puts them at increased risk of stunting, wasting and anaemia. Previous studies have suggested this relationship but have suffered from confounding by socioeconomic status. Comparison with non-working siblings reduces the chances of socioeconomic status confounding. PMID- 11856187 TI - Can maternal depression increase infant risk of illness and growth impairment in developing countries? AB - Despite relative improvement in living conditions and availability of modern healthcare, infant mortality rates continue to be very high in many developing countries. High rates of depression have also been reported in women in these countries. The continuous care and attention of children is a demanding task, and poor physical or mental health in mothers might be expected to have adverse consequences on their children's health, nutrition and psychological well-being. Review of published literature reveals very little research in developing countries on the association between poor mental health in mothers and the subsequent physical well-being of their children. We hypothesize that the level of care provided by mothers with depression may put their infants at higher risk of infection and impaired growth, compared with infants of mothers without depression. We outline approaches to test such a hypothesis in a developing country, and discuss its implications. PMID- 11856188 TI - Environmental barriers in mainstream schools. AB - Research on students with disabilities in mainstream schools often focuses on the students' personal abilities rather than on the establishment itself. To promote inclusive education, the environmental prerequisite for participation has to be explored also. The aim of this study was to identify the barriers to participation in Swedish mainstream schools, from the personal perspective of students with physical disabilities. The study also investigated how gender, diagnosis, level of mobility, academic years and availability of an assistant were related to student-environment fit. A total of 34 students with physical disabilities, aged between 10 and 19 years, participated in the study. The students were assessed by 'The school-setting interview'. Results show that two thirds of the students experienced barriers to participation in both the physical and the social environment. A majority of the barriers originated from the way in which school activities were organized and carried out in schools. Failure to provide adequate environmental adjustments resulted in restricted participation or exclusion from some of the activities in class. Older students experienced significantly more barriers than younger ones because the school organization was less favourable. The results suggest that the way in which activities are organized in school is the area in need of most improvements to promote participation of students with physical disabilities. PMID- 11856189 TI - Communicating a diagnosis of developmental disability to parents: multiprofessional negotiation frameworks. AB - It is a hard task for professionals to give and parents to receive the bad news about a child's developmental disability. This study describes how findings about four preschool children, with difficulties suspected to lie within the autistic spectrum, were negotiated with parents by two multiprofessional groups, one in a Medical (Site M) and one in an Education setting (Site E). Each assessment was undertaken over half a day, the professionals jointly interacting with the child and family. All assessment discussions were audiotaped and each participant was interviewed after the assessment. Conversation analysis showed the activation of three major social-interaction frameworks: professionals at both sites applied a parent-friendly frame, but this was complemented by a hopeful-diagnostic formulation frame at Site M, and a defocusing-of-bad-news frame at Site E. PMID- 11856190 TI - Coping of parents with physically and/or intellectually disabled children. AB - OBJECTIVE: The aim was to clarify how families with physically and/or intellectually disabled children cope, what kind of coping strategies they use and how the families with good and poor coping capacities differ. METHODS: The parents of eight children (aged 8-10 years) with physical and/or intellectual disability were interviewed twice, and the data elicited in these interviews were analysed qualitatively using the grounded theory method. RESULTS: Information and acceptance, good family co-operation and social support were related to the coping strategies most frequently used. Half of the families seemed to have found successful ways of coping, whereas another half had major problems. There were five main domains in which the high- and low-coping families differed most from each other: (1) parents' initial experiences; (2) personal characteristics; (3) effects of the child's disability on family life; (4) acting in everyday life; and (5) social support. CONCLUSION: The findings can be utilized in developing supportive activities for families with disabled children. By recognizing the coping strategies used by the family, professionals and service providers can find the right ways to support their adaptation. As the role of physicians, nursing staff and other professionals in this process is very important, more attention should be attached to the collaboration between these groups, to enable them to view the situation from the perspective of the whole family. PMID- 11856191 TI - Disability in children from different ethnic populations. AB - We report on the prevalence of severe neurodisability in children in the Southern Derbyshire Health Authority from different ethnic groups. Information was obtained from the health records of children at the Child Development Centre (CDC), and analysed according to ethnic group. There were 53 in the Pakistani group, 20 in the Indian group and 764 in a mixed group, of which 95% were of European origin. It was estimated that all children with severe disability in the area of the Health Authority had notes at the CDC, except for 10% of the mixed group living on the periphery. The numbers of children with different disabling conditions were recorded, together with a measure of the level of individual disability; the 'Disability Scores'. We also noted if the condition was genetic or chromosomal in origin. Pakistani children showed a higher prevalence than the other groups of severe learning disorder, severe and profound hearing loss and severe visual problems. They also had a slightly increased prevalence of autism and cerebral palsy. Conversely, they showed a lower prevalence of language disorder. Disability scores for Pakistani children attending the CDC were higher than for other groups. Genetic disease causing disability was 10 times more common in the Pakistani children than other ethnic groups. Disability is more common in Pakistani children probably as a result of the cultural practice of consanguineous marriages. This community needs special help for disabled children, and their families, for general support and appropriate genetic counselling. PMID- 11856192 TI - Not by bread alone': impact of a structured 90-minute play session on development of children in an orphanage. AB - BACKGROUND AND OBJECTIVES: In developing countries, caring for the large number of babies in orphanages is very hard work. Whereas the physical needs of most of the children are met, play often gets neglected. Studies have repeatedly shown that babies in such institutionalized settings suffer from severe psychomotor retardation. The aim of this project was to develop an intervention programme of structured play. We hypothesized that such an intervention would result in acceleration of psychosocial development in otherwise healthy institutionalized children. DESIGN: Prospective longitudinal. SETTING: Mother Teresa's Orphanage, run by Missionaries of Charity. SUBJECT AND METHODS: All 30 children in the orphanage aged 6 months-2.5 years, were assessed for their Motor, Mental and Social Quotients, using the Indian adaptation of Bailey's Scale of Infant Development(DASII) and the Vineland's Social Maturity Scale. A structured 'Regime of Play' was then built into the routine of the orphanage. A repeat developmental assessment was performed at the end of 3 months to assess the impact. RESULTS: Out of the original cohort of 30, 19 children were available for post intervention assessments. The remainder were adopted before their assessments. Their mean Motor Quotient rose from 63.7 to 81.7, mean Mental Quotient rose from 65.8 to 89.6 and the mean Social Quotient rose from 61.9 to 91.3, a gain of 18, 23 and 30 points respectively (p < 0.0001). There was also an overall change in the environment of the orphanage. Children became more active, playful, responsive and independent. Contrary to what caretakers assumed, their workload actually decreased. The responsiveness in the children awakened as a result of play, acted as a positive feedback for caretakers to continue the play sessions. CONCLUSIONS: This study shows that short daily sessions of play can significantly improve the development of children in such institutions. It is vital to remember that children grow 'Not by Bread Alone'. PMID- 11856193 TI - Secondary school accident reporting in one education authority. AB - BACKGROUND: Secondary schools appear to have very different accident rates when they are compared on the basis of accident report returns. The variation may be as a result of real differences in accident rates or different reporting procedures. This study investigates accident reporting from secondary schools and, in particular, the role of the school nurse. METHODS: Accident form returns covering a 2-year period were collected for statistical analysis from 13 comprehensive schools in one local education authority in Wales. School sites were visited in the following school year to obtain information about accident records held on site and accident reporting procedures. RESULTS: The main factors determining the number of school accident reports submitted to the education authority relate to differences in recording and reporting procedures, such as the employment of a nurse and the policy of the head teacher/safety officer on submitting accident returns. Accident and emergency department referrals from similar schools may show significant differences in specific injuries and their causes. CONCLUSIONS: The level of school accident activity cannot be gauged from reports submitted to the education authority. Lack of incentives for collecting good accident data, in conjunction with the degree of complacency in the current system, suggest that future accident rates and reporting activity are unlikely to change. PMID- 11856206 TI - Helicobacter pylori infection. PMID- 11856194 TI - I've come for his throat': roles and identities in doctor-parent-child communication. AB - Previous studies on doctor-parent-child communication at the general practitioner's surgery showed that the GP and the parent differ fundamentally in the way they enable or constrain child participation. The question how to explain these differences is at the core of the present study. The aim is to describe how the three participants display their orientation to their institutional roles and identities; how they collaboratively co-construct the course of action; and how these discursive constructions structure the ongoing interaction. A qualitative analysis of 106 videos shows that although GP and parent initially show incongruent orientations toward child participation, in the further course of the encounter all three participants jointly establish a situation in which child participation appears to be rather an exception. It is concluded that parental speaking for the child is, in a way, institutionally co-constructed; parents take their responsibility, which is hardly ever questioned by children, and GPs ratify this behaviour by refraining from meta-communicative comments and by aligning with the parent in the course of the interaction. The results are discussed in terms of enabling child participation and implications for medical practice. PMID- 11856207 TI - The antibacterial effects of ciprofloxacin and trovafloxacin against Streptococcus pneumoniae in an in vitro dynamic model. AB - OBJECTIVE: To determine whether the newer fluoroquinolone antibiotics such as trovafloxacin posses enhanced activity against Gram-positive organisms, including Streptococcus pneumoniae, because the clinical activity of older quinolones against pneumococci has been questioned. METHODS: In this study, the bactericidal activities of ciprofloxacin and trovafloxacin against six strains of penicillin resistant and -sensitive strains of Streptococcus pneumoniae were compared using an in vitro model that simulates human pharmacokinetics. Ciprofloxacin was administered at 750 mg every 12 h, higher than the usual daily dose of 500 mg twice a day. Trovafloxacin was administered at 300 mg every 24 h for the six strains and at 200 mg every 24 h for three of the strains. RESULTS: The single 300-mg dose of trovafloxacin killed five of the six strains in 4 h, with no bacterial regrowth. Ciprofloxacin reduced the initial inoculum by 3--5 logs by 24 h. Although the 300-mg dose of trovafloxacin more rapidly eradicated susceptible strains, the activity of trovafloxacin at 200 mg every 24 h was similar to that of ciprofloxacin at 750 mg every 12 h against the three strains tested. CONCLUSION: Trovafloxacin (and ciprofloxacin at high doses) eradicates susceptible strains of pneumococci in an in vitro dynamic model. PMID- 11856208 TI - Hemolytic uremic syndrome in Belgium: incidence and association with verocytotoxin-producing Escherichia coli infection. AB - OBJECTIVE: To evaluate the incidence of hemolytic uremic syndrome (HUS) in Belgium and to determine the role of verocytotoxin-producing Escherichia coli O157:H7 and other serotypes (non-O157 VTEC). METHODS: Twenty-two centers, including the seven university hospitals, registered prospectively all cases of HUS; they collected clinical samples for isolation of VTEC strains and serum for detection of specific O-lipopolysaccharide antibodies. RESULTS: Forty-seven cases of HUS (including five incomplete cases) were recorded. Three cases were seen in non-residents. The incidence of complete HUS in Belgian residents was 4.3 cases/100 000 in children <5 years old, 1.8 cases/100 000 when all children <15 years were considered, and 0.42/100 000 when patients of all ages were taken into account. By combining bacteriologic and serologic results, evidence of VTEC infection was obtained in 64% of the patients, mainly but not exclusively in children with prodromal diarrhea. The 13 VTEC isolates belonged to serotypes O157:H7 (nine isolates), O26:H11, O121:H---, O145:H--- and O172:H--- (one each) and all produced VT2 (+VT2vh-a in three O157 strains) and were positive for the eaeA gene. CONCLUSIONS: The incidence rate found in this study and the high mortality and morbidity linked with this syndrome warrant further registration of pediatric and post-diarrheic adult HUS cases and also examination of stools for both O157 and non-O157 VTEC strains. For effective prevention of this disease, further study of the serotypes and accessory virulence factors associated with HUS is needed. PMID- 11856209 TI - Quantitation of human cytomegalovirus DNA in peripheral blood leukocytes of heart transplant recipients: relationship with pp65 antigenemia and with antiviral therapy. AB - OBJECTIVE: To retrospectively determine DNA levels in blood polymorphonuclear leukocytes (PMNLs) of 21 heart transplant patients who suffered from HCMV infection and who were monitored by the antigenemia assay (pp65 test) during follow-up, by use of a quantitative competitive polymerase chain reaction (PCR) assay for human cytomegalovirus (HCMV) DNA. METHODS: Quantitation of HCMV DNA by PCR was expressed as genome equivalents (GE) per 200 000 PMNLs. RESULTS: Ten patients experienced symptomatic HCMV infection (five primary infections and five reactivations) with mild symptoms and received ganciclovir treatment, whereas 11 asymptomatic HCMV infections were not treated. Therapy was discontinued when a 90% reduction of the pretreatment antigenic load was achieved in a symptomless patient. The mean HCMV DNA and antigenic loads were significantly higher in symptomatic than in asymptomatic patients: 4.6 x 105 plus minus 4.7 x 105 GE and 1.1 x 104 GE (p<0.0001) and 390 plus minus 350 versus 25 plus minus 12 pp65 positive PMNLs (p<0.0001), and in primary than in secondary infections (583 plus minus 403 pp65-positive PMNLs versus 85 plus minus 111, p=0.002 and 5.2 x 105 plus minus 5.2 x 105 GE instead of 1.5 x 105 plus minus 3.2 x 105 GE, p=0.02). A single course of 14--21 days of ganciclovir caused a marked decrease of HCMV DNA and antigenemia in eight of 10 patients in whom a 90% reduction of the antigenic load correlated with a 98% DNA reduction of the pretreatment levels. In two primary infections, a 90% antigenic reduction was achieved by 21 days of ganciclovir treatment, but those data only correlated with a DNA load reduction of 28% and 60% of the pretreatment levels. Fifteen and 12 days later, respectively, the two patients relapsed and underwent a second ganciclovir course, at the end of which a 90% reduction of the antigenic load correlated with a >98% DNA drop. GCV was discontinued and the patients recovered completely. In those two patients we retrospectively found persistent high DNA levels before the second ganciclovir course, whereas the antigenic load slowly increased after an apparent reduction. CONCLUSIONS: Our data suggest that: (1) DNA levels have the same trend as the pp65 antigen test---they are significantly higher in symptomatic and in primary HCMV-infected patients than in asymptomatic patients and those with secondary infection; (2) a 90% antigenic load reduction from the pre-treatment level may be a less reliable predictor of the efficacy of anti-HCMV therapy than DNA load, at least in primary infection, in which a much higher viral load and much more severe disease are present; and (3) a DNA load reduction of >98% of the pretreatment value is required for therapeutic success. PMID- 11856210 TI - In vitro assessment of colistin's antipseudomonal antimicrobial interactions with other antibiotics. AB - OBJECTIVE: To study the interactions of colistin (MIC 2 mg/L) at concentrations of 0.5 and 5 mg/L with ceftazidime (1 and 75 mg/L, MIC 0.5 mg/L), aztreonam (1 and 30 mg/L, MIC 0.12 mg/L), meropenem (1 and 25 mg/L, MIC 0.03 mg/L), gentamicin (1 and 10 mg/L, MIC 2 mg/L), piperacillin (5 and 100 mg/L, MIC 4 mg/L) and ciprofloxacin (0.25 and 4 mg/L, MIC 1 mg/L) using a representative strain of Pseudomonas aeruginosa isolated from a cystic fibrosis patient. METHODS: The method used was a bacterial time kill curve with single agents and combinations. Using inocula of 106 CFU/mL, multiple sampling was performed over 6 h and in triplicate. The AUBKC of the time versus viable count curve, with single agents and combinations of agents, was taken as the endpoint for comparison. RESULTS: For colistin plus ceftazidime, colistin plus aztreonam, colistin plus meropenem and colistin plus ciprofloxacin, the pattern was for all the combinations (high or low concentrations) to produce smaller AUBKCs than single agents. In experiments using a bacteriostatic agent such as ceftazidime, the AUBKCs (log CFU/uL per h) for colistin 0.5 mg/L or 5 mg/L alone were 32.3+/-0.8 or 12.7+/ 0.5, and for ceftazidime 1 mg/L or 75 mg/L alone they were 24.3+/-1.5 or 20.9+/ 2.7. Combinations of colistin 0.5 mg/L plus either ceftazidime 1 mg/L or 75 mg/L produced AUBKCs of 23.8+/-1.8 or 16.1 mg/L. Combinations of colistin 5 mg/L plus ceftazidime 1 mg/L or 75 mg/L produced AUBKCs of 12.2+/-0.8 or 8.7+/-1.0. The AUBKCs for colistin 5 mg/L plus 75 mg/L are significantly smaller than those for the single agents, indicating synergy. In experiments using the bactericidal agent ciprofloxacin, the AUBKCs (log CFU/mL per h) for colistin 0.5 mg/L or 5 mg/L alone were 33.6+/-1.9 or 11.2+/-2.4, and for ciprofloxacin 0.25 mg/L or 4 mg/L alone they were 32.8+/-1.3 or 5.0+/-0.7. Combinations of colistin 0.5 mg/L plus either ciprofloxacin 0.25 mg/L or 4 mg/L produced AUBKCs of 32.2+/-0.9 or 4.3+/-1.4. Combinations of colistin 5 mg/L plus ciprofloxacin 0.25 mg/L or 4 mg/L produced AUBKCs of 10.7+/-1.5 or 4.2+/-0.6. Although combination AUBKCs were smaller than those for single agents, in no case did this reach statistical significance (p<0.05). CONCLUSIONS: These studies indicate that addition of colistin to other antipseudomonal drugs tends to produce smaller AUBKCs and hence greater killing of Pseudomonas aeruginosa than monotherapy. PMID- 11856211 TI - Bacterial and epidemiologic study of the resistance to oxyiminocephalosporins in Escherichia coli in a French hospital. AB - OBJECTIVE: To understand the mechanisms and epidemiology of resistance to oxyiminocephalosporins in Escherichia coli over a 2-year period in a French hospital. METHODS: Forty-four strains, resistant or intermediately resistant to one of the oxyiminocephalosporins or aztreonam, were collected from 35 patients. MIC determinations were carried out for the 44 isolates using a panel of beta lactam antibiotics, and characterization of the beta-lactamases they produced by isoelectric focusing and catalytic activity measurement. Extended-spectrum beta lactamase production was studied by use of the double disk diffusion test. Conjugation experiments were used to search for plasmidic cephalosporinase. An epidemiologic study was then performed, by use of molecular typing of the strains with an ERIC-PCR method and a case-control analysis. RESULTS: Less than 1% of all the E. coli isolates at our hospital showed decreased susceptibility to oxyiminocephalosporins. Only three of the 44 isolates showed synergy between clavulanate and a third-generation cephalosporin and produced an extended spectrum beta-lactamase. For the other strains, a beta-lactamase with a highly basic isoelectric point was detected. Spectrophotometric measures confirmed that most of these isolates were AmpC hyper-producers. No plasmidic cephalosporinase could be detected by conjugation experiments. Molecular typing showed all isolates to be different, except for two strains isolated in two patients of the same hospital unit, and for the repeated isolates of some patients. When 20 case patients were compared to 40 randomly selected control patients, prior receipt of an antimicrobial and more specifically of a beta-lactam agent was significantly associated with case patients. CONCLUSIONS: Although it appears to be very rare, the resistance to broad-spectrum cephalosporins needs our attention, because of the high frequency of E. coli infections and beta-lactam use in their treatment. PMID- 11856212 TI - Experimental Yersinia enterocolitica 0:3 arthritis in rats. PMID- 11856213 TI - Kidney and pancreas transplantation: postoperative infectious complications. PMID- 11856214 TI - In vitro activity of linezolid and eperezolid against anaerobic bacteria. PMID- 11856215 TI - Identification of methicillin-resistant Staphylococcus aureus by latex agglutination kits: performance with epidemic strains (EMRSA) and strains causing problems with latex agglutination methods. PMID- 11856216 TI - Urethritis caused by Neisseria meningitidis serogroup C. PMID- 11856217 TI - Further increase in ciprofloxacin-resistant Campylobacter jejuni/coli in Styria, Austria. PMID- 11856218 TI - Towards better surveillance of bacterial antibiotic resistance. PMID- 11856219 TI - Vancomycin-resistant Enterococcus faecium derived from animals colonizing the human gut: the missing evidence? PMID- 11856220 TI - Prevalence of yersinia plasmid-encoded outer protein (Yop) class-specific antibodies in multitransfused Greek patients with thalassemic syndromes. AB - OBJECTIVE: To evaluate the prevalence of class-specific antibodies (G, A, M) to Yersinia enterocolitica plasmid-encoded outer proteins (Yops), in a closely followed multitransfused population of patients with thalassemia. METHODS: Sera from 408 beta-thalassemic patients and 386 healthy blood donors used as controls were analyzed with the enzyme-linked immunosorbent assay (ELISA) for IgG, IgA and IgM antibodies to yersinia outer proteins. The Yop antigen for the ELISA was prepared using a plasmid-bearing wild-type strain of Y. enterocolitica of serotype O:8. RESULTS: Anti-Yop IgG antibodies were detected in 84 out of 408 beta-thalassemic patients (20.6%) compared with only eight out of 386 (2.1%) healthy blood donors. None of the sera of either group was positive for anti-Yop IgA or IgM antibodies. On evaluating patients with registered clinical and laboratory signs of a previous yersinia infection in the period from 1978 to 1996, we found that those with a positive agglutination test for Y. enterocolitica infection at the time of manifestation showed a higher rate of persisting IgG seropositivity to Yops than those with positive culture and clinical signs only. A significant percentage (9.49%) of the seropositive patients had no registered data of a past Y. enterocolitica infection. There was remarkable persistence of anti-Yop IgG antibodies in the thalassemic population, even in patients infected during the early years of our study period (1978--80). CONCLUSIONS: The results suggest that the determination of class-specific antibodies to Yops, which are specific antigens for the pathogenic yersiniae (Y. enterocolitica, Y. pseudotuberculosis and Y. pestis), in addition to its usefulness in the diagnosis of infection, will be a very sensitive and specific index for epidemiologic studies. PMID- 11856221 TI - Current status of taxonomic groups of oral streptococci in endocarditis. Can virulence factors discriminate between endocarditis and non-endocarditis strains? AB - OBJECTIVE: Infective endocarditis is frequently caused by oral streptococci, especially Streptococcus sanguis. In this group, many strains have recently been reclassified on the basis of new taxonomic schemes. The purpose of this study was to classify oral streptococci from patients with infective endocarditis and, further, to assess the importance of specific virulence factors for the development of streptococcal endocarditis. METHODS: Twenty-eight previously identified and 10 new streptococcal isolates from infective endocarditis were classified according to Kilian et al (1989) and compared to 30 streptococcal isolates from the oral cavities of periodontal patients without endocarditis. Subsequently, surface hydrophobicity was assessed by hydrophobic interaction chromatography, production of extracellular dextran was determined by precipitation, and non-specific proteolytic activity was evaluated by determination of hydrolysis of gelatin, and casein-precipitating activity. RESULTS: Eight streptococcal species were represented in the endocarditis isolates. Most strains were highly hydrophobic and none showed non-specific proteolytic activity. Dextran was produced with similar frequency in endocarditis and non-endocarditis isolates. CONCLUSIONS: The present study showed that infective endocarditis may be caused by a variety of oral streptococcal species. The possible virulence factors investigated were found in the same proportions in endocarditis and non-endocarditis isolates, and thus did not seem to be crucial for development of endocarditis. PMID- 11856222 TI - Determination of cytomegalovirus DNA load for monitoring of cytomegalovirus disease and antiviral treatment in solid organ transplant patients, comparing limiting-dilution PCR and hybrid capture assay with cytomegalovirus isolation. AB - OBJECTIVE: To improve the identification of patients at risk of developing cytomegalovirus (CMV) disease. MATERIALS AND METHODS: In a prospective study of 50 kidney or liver transplant patients who developed fever, 133 EDTA blood samples were analyzed, using two tests to measure CMV DNA: a 10-fold limiting dilution of an extract of 2 million leukocytes for CMV PCR, and a CMV hybrid capture assay. Both tests were compared with virus isolation, using an equivalent amount of leukocytes as a base for all three tests. RESULTS: The limiting dilution CMV PCR and the hybrid capture assay presented relatively similar changes of sensitivity and specificity at different CMV DNA concentrations. The kinetics of the positive and negative predictive values were also comparable. A higher CMV DNA load corresponded to an increased risk of developing CMV disease. Furthermore, an increase in the endpoint dilution of a positive CMV PCR also corresponded to more severe disease. After antiviral treatment, the CMV PCR decreased by at least 100-fold (2 log10) in 10 cases and by 10-fold (1 log10) in five cases. Thus, there was a decrease in 15 of 18 (83%) patients. Similarly, with the hybrid capture assay, the amount of CMV DNA decreased about 100-fold in five patients and decreased by about 0.5 genome equivalents in five cases, i.e. in 10 of 12 (83%) patients. CONCLUSION: Both methods proved clinically useful for detecting patients at risk of developing CMV disease and for monitoring antiviral treatment in solid organ transplant patients. PMID- 11856223 TI - A new biotyping method for Streptococcus mutans with the API ZYM system. AB - OBJECTIVE: To test a new system for the biotyping of Streptococcus mutans, based on the measurement of enzyme activity, and to investigate the relationship between biotype and in vitro susceptibility to seven clinically useful antibiotics. METHODS: In total, 160 oral isolates of S. mutans were classified into different biotypes with the APIZYM test for enzyme activity, excluding results that were positive or negative in >80% of the strains. The susceptibility of all 160 strains to amoxycillin, cefazolin, erythromycin, clindamycin, vancomycin, teicoplanin and imipenem was tested by dilution in a solid medium. Statistical analysis of susceptibility (mean minimum inhibitory concentrations (MICs)) was based on chi-squared tests. RESULTS: Eight different biotypes (1-8) were identified on the basis of three kinds of enzyme activity: valine aryl amidase, acid phosphatase and alpha-galactosidase. Biotype 5 was found to be the most common. The mean MIC values showed strains belonging to biotype 4 to be the most susceptible to amoxycillin, cefazolin and erythromycin, whereas biotype 1 was the least susceptible to teicoplanin. CONCLUSIONS: The proposed biotyping method, which is relatively fast and simple to perform, provided reproducible results, and may contribute to clinically effective treatment of S. mutans infections. PMID- 11856224 TI - Polymerase chain reaction, with sequencing, as a diagnostic tool in culture-- negative bacterial meningitis. AB - OBJECTIVE: To evaluate the feasibility of using 16S rDNA universal primer PCR (followed by sequencing) and 65-kDa heat shock Mycobacterium tuberculosis protein gene PCR as a method to determine a bacterial etiology in culture---negative cerebrospinal fluid (CSF) samples. METHODS: One hundred and forty-nine CSF samples from 128 patients were processed. DNA was extracted from the CSF samples and amplified with the eubacterial 16S rDNA primers P11E and P13B, and with the 65-kDa heat shock protein gene mycobacterial primers. The amplicons were identified by sequencing and specific oligoprobe hybridization. RESULTS: Overall, a microbiological diagnosis was made in 11 of 125 ultimately culture-negative cases. The use of 65-kDa heat shock protein gene PCR was needed to improve the diagnosis of tuberculous meningitis; in four patients, prospectively studied, the outcome of antituberculous therapy could also be followed. CONCLUSIONS: In culture-negative bacterial meningitis it is possible to improve the microbiological diagnosis by use of 16S rDNA amplification and sequencing, together with amplification of a more specific gene in mycobacteria. PMID- 11856225 TI - Intestinal colonization of a human subject by vancomycin-resistant Enterococcus faecium. AB - OBJECTIVE: To study the ability of two strains of vancomycin-resistant Enterococcus faecium to colonize the human intestine. METHODS: A single human subject ingested separately two strains of vancomycin-resistant E. faecium isolated from a pig and a chicken. The feces were cultured on selective medium. Prior to ingestion no vancomycin-resistant cocci were present in the feces. Ingestion of 104-105 CFU resulted in either no colonization or isolation only after enrichment. Ingestion of 107 CFU of one strain resulted in colonization for a period of nearly 3 weeks, with fecal counts at times in excess of 106 CFU/g. Ingestion of similar numbers of the other strain and reingestion of the first strain resulted in excretion in the feces for much shorter periods. When the fecal count of the ingested strains was greater than 104-105 CFU/g, the strains were isolated from swabs taken from perianal skin. CONCLUSIONS: Vancomycin resistant E. faecium strains from pigs and poultry are able to colonize the human gut and the perianal skin. PMID- 11856226 TI - Pasteurella multocida septicemia and meningitis in an infant. PMID- 11856227 TI - Outbreak of glycopeptide-resistant Enterococcus faecium in a neonatal unit. PMID- 11856229 TI - Diagnostic value of Gram stain and culture of sputum and endotracheal aspirates in bacteremic pneumococcal pneumonia. PMID- 11856228 TI - Human retrovirus-5 and Sjogren's syndrome. PMID- 11856230 TI - Molecular epidemiologic analysis of multiresistant Klebsiella pneumoniae isolates from a pediatric hospital. PMID- 11856232 TI - The ability of the Dry Spot Staphytect Plus test, in comparison with other tests, to identify Staphylococcus species, in particular S. aureus. PMID- 11856231 TI - Pneumonia due to Stomatococcus mucilaginosus in an AIDS patient: case report and literature review. PMID- 11856233 TI - An archival case of clinically significant infection in a surgical patient by enterotoxigenic Bacteroides fragilis: PCR amplification of the metalloprotease enterotoxin gene from non-viable lyophilizates. PMID- 11856234 TI - Antimicrobial use in animals and its consequences for human health. PMID- 11856235 TI - Prevalence of hepatitis C virus antibodies in a clinic-based group of Italians from one geographic area. AB - OBJECTIVE: To evaluate the prevalence of anti-HCV antibodies using subjects hospitalized in surgical departments and medical wards, and out-patients; secondly, to assess the evidence for developing chronic hepatitis in subjects positive for anti-HCV when compared with those with hepatitis B virus (HBV). METHODS: 21 888 serum samples from 18 380 subjects were investigated for anti-HCV antibodies using second and third generation immunoenzymatic assays. Some of these subjects were hospitalized patients and some were out-patients. RESULTS: THE STUDY SHOWED A 12.8&PERCNT; OVERALL ANTI-HCV PREVALENCE RATE WITH SIGNIFICANT DIFFERENCES BETWEEN OUT-PATIENTS (16.5&PERCNT;) OR SUBJECTS HOSPITALISED IN MEDICAL WARDS (16&PERCNT;) AND IN-PATIENTS IN SURGICAL DEPARTMENTS (7.7&PERCNT;). THE THIRD GROUP INCLUDED ASYMPTOMATIC SUBJECTS OVER TWENTY YEARS OLD WHOSE SERA WERE TESTED FOR ANTI-HCV ANTIBODIES AS PART OF ROUTINE PREOPERATION SCREENING AND NOT ON CLINICAL SUSPICION. HENCE, THIS GROUP, TOO, CAN BE CONSIDERED AS REPRESENTATIVE OF THE GENERAL POPULATION, AND THE PREVALENCE OF ANTI-HCV ANTIBODIES OBSERVED AMONG THEM AS THE PREVALENCE OF ANTI-HCV ANTIBODIES IN THE GENERAL POPULATION IN A NORTHERN ITALIAN AREA. THE DATA, FOLLOWING A CONFIRMATORY TEST (RIBA) ON POSITIVE SAMPLES, WERE ANALYSED FOR THEIR POSITIVITY TO DIFFERENT ANTIGENS (THE SIMULTANEOUS PRESENCE OF ANTIBODIES TO THE C-100, C-33 AND C-22 ANTIGENS), AS AN INDEX OF DEVELOPING CHRONIC VIRAL ACTIVITY. THIS WAS OBSERVED IN 63.4&PERCNT; OF POSITIVE PATIENTS FROM SURGICAL DEPARTMENTS: CONCLUSIONS: There is a large proportion of the asymptomatic population which could be chronically infected. PMID- 11856236 TI - Resistance to ciprofloxacin in non-typhoidal salmonellas from humans in England and Wales-the current situation. AB - OBJECTIVE: To report on the prevalence of isolates with ciprofloxacin resistance in non-typhoidal salmonellas from humans in England and Wales in 1997. METHODS: All non-typhoidal salmonellas referred to the Laboratory of Enteric Pathogens in 1997 were screened for resistance to ciprofloxacin at 0.125 and 1.0 mg/L and nalidixic acid at 16 mg/L, and results were compared to those for 1994. Full minimal inhibitory concentrations (MICs) of these antimicrobials were also determined for a selection of isolates resistant to ciprofloxacin at 0.125 mg/L but sensitive at 1.0 mg/L, and for all isolates resistant at 1.0 mg/L. RESULTS: Since 1994 there have been increases in the occurrence of resistance to ciprofloxacin (MICs: 0.25-1.0 mg/L) in Salmonella enterica serotypes Enteritidis, Typhimurium, Virchow and Hadar. Of particular importance have been increases in the occurrence of resistance in multiresistant S. Typhimurium DT 104, and also in S. virchow, a serotype with a propensity for causing extraintestinal infections in humans. High-level resistance (MIC >/=2.0 mg/L) was uncommon and was identified in only a few strains, all from patients with a history of recent foreign travel. CONCLUSIONS: There is a strong temporal association between increases in the occurrence of ciprofloxacin resistance in Salmonella serotypes Typhimurium, Virchow and Hadar from humans in England and Wales and with the licensing for use in food animals in the UK of the related fluoroquinolone antibiotic enrofloxacin; in contrast, for S. enteritidis ciprofloxacin resistance was most common in a phage type associated with foreign travel. It is hoped that recent recommendations for the use of fluoroquinolone antimicrobials in food animals in the UK will result in a reduction in the occurrence of resistance to ciprofloxacin in zoonotic salmonellas causing infections in humans. PMID- 11856237 TI - Wound infections following posterior spinal instrumentation for paralytic scoliosis. AB - OBJECTIVES: To investigate in a prospective study the infection rate, the bacteriology and the outcome of wound infections following posterior spinal instrumentation in children and young adolescents during 1993-96. METHODS: Fifty patients, who underwent spinal instrumentation because of paralytic scoliotic deformity, were followed by the surgeon, the clinical microbiologist and the infection control nurse. In those patients with clinical and laboratory findings suggesting wound infection, multiple swabs and tissue biopsies were obtained from deep within the infected wound and were cultured on appropriate media. Microorganisms were identified by conventional methods. RESULTS: Ten of 50 patients (20%) developed early deep wound infections, most of them polymicrobial, 3-9 days after the operation. Coagulase-negative staphylococci (70.0%), Enterobacteriaceae (17.3%), anaerobes (5.4%) and Staphylococcus aureus (3.7%) were isolated from the wound specimens. S. epidermidis strains were the predominant isolates. All coagulase-negative staphylococci were multiresistant to beta-lactams, aminoglycosides, fusidic acid and co-trimoxazole, while most of them were susceptible to rifampicin and quinolones. All wounds healed uneventfully with aggressive debridement and prolonged antimicrobial therapy. CONCLUSIONS: Postoperative wound infection is a significant complication of spinal instrumentation. Multiresistant coagulase-negative staphylococci are the predominant pathogens. Successful treatment includes wound debridement and prolonged antimicrobial therapy. PMID- 11856238 TI - Pharmacodynamic effects of amikacin, ciprofloxacin and imipenem on growing and non-growing Escherichia coli and Pseudomonas aeruginosa. AB - OBJECTIVE: To compare control-related effective regrowth times (CERTs) and postantibiotic effects (PAEs) of amikacin, ciprofloxacin and imipenem on growing and non-growing Escherichia coli and Pseudomonas aeruginosa. METHODS: CERTs and PAEs of amikacin, ciprofloxacin and imipenem were determined with bioluminescence assay of bacterial ATP and viable counts. RESULTS: Negative viable count PAEs of amikacin and imipenem occurred on growing bacteria, but bioluminescence PAEs were positive. CERTs were equal with both methods. Amikacin and ciprofloxacin induced long, concentration-dependent CERTs on growing and non-growing cultures. Amikacin (32 mg/L) prevented regrowth of E. coli and induced a CERT of 6.0 h on P. aeruginosa; corresponding CERTs on non-growing bacteria were 3.4 h and 3.3 h, respectively. Ciprofloxacin (8 mg/L) prevented regrowth of both strains in growing cultures and induced CERTs of 5.1 h on non-growing E. coli and 13.3 h on P. aeruginosa. Imipenem induced a concentration-dependent CERT on growing bacteria and no CERT on non-growing cultures. Imipenem (16 mg/L) induced a CERT of 5.3 h on growing P. aeruginosa and 3.2 h on E. coli. CONCLUSION: Amikacin and ciprofloxacin induced strong pharmacodynamic effects on growing and non-growing E. coli and P. aeruginosa, while imipenem was only effective on growing cultures. PMID- 11856239 TI - Clinical and molecular epidemiology of hospital Enterococcus faecalis isolates in eastern France. AB - OBJECTIVE: To report on the occurrence of Enterococcus faecalis hospital isolates obtained during 1 year in hospitals in the Franche-Comte region of France. METHODS: Clinical isolates of E. faecalis of different antibiotic susceptibility phenotypes from hospitalized patients were characterized by pulsed-field gel electrophoresis. Patients with positive cultures were investigated by three case control studies to identify risk factors for colonization/infection. RESULTS: The crude incidence of colonization/infection was 2.37%, and 4-day and 7-day colonization rates after admission were 10.0% and 6.36%, respectively. The rates of high-level resistance to kanamycin (HLKR) and to gentamicin (HLGR) were 47.1% and 7.1%, respectively. No isolate was resistant to glycopeptides or produced beta-lactamase. The 209 hospital isolates obtained during the study yielded 98 major DNA patterns, of which two were major epidemic patterns including HLKR isolates. No single factor was significantly associated with colonization/infection by HLKR isolates. The length of hospitalization before isolation was associated with colonization by HLGR isolates. CONCLUSIONS: The isolation frequency of E. faecalis strains with acquired resistance to aminoglycoside antibiotics, and the wide dissemination of resistant strains with characteristics that allow them to persist and spread, argue for further large prospective surveys of clinical isolates of E. faecalis in hospitals. PMID- 11856240 TI - Experimental endocarditis in the rat secondary to septic arthritis induced by Staphylococcus aureus. AB - OBJECTIVE: To develop a modified model for experimental infective endocarditis (IE) in the rat. The goal was to induce a primary infectious focus in the temporomandibular joint (TMJ) of a rat. Hematogenous translocation of the bacteria to the traumatized aortic valve was desired. METHODS: Catheterization of the right carotid artery through the aortic valve was performed 7 days after induction of arthritis, which was done by intra-articular injection of glucocorticosteroid (triamcinolone acetonide, 1 mg) and intravenous challenge with 107 CFU Staphylococcus aureus. RESULTS: TMJ arthritis could be induced by intra-articular triamcinolone acetonide followed by intravenous bacterial challenge. Joints not given glucocorticosteroid were not affected. Only rats with arthritis developed IE subsequent to catheterization as a result of bacteremia generated from the arthritis. CONCLUSIONS: The present model may serve as a complement to the conventional method for induction of IE, in which a high intravenous challenge has to be given. In the present model, IE was instead the result of a continuous low level of bacteremia from an infectious focus in the TMJ. This model mimics the natural development of IE in patients, and may assist as a setting for prophylactic and therapeutic trials. PMID- 11856241 TI - Increased intracranial pressure and hydrocephalus in a teenager. PMID- 11856242 TI - Bacteroides species highly resistant to metronidazole: an emerging clinical problem? PMID- 11856243 TI - A promising cultural-biomolecular method for rapid isolation and identification of mycobacteria. PMID- 11856244 TI - Rapidly progressive tricuspid valve endocarditis caused by Capnocytophaga canimorsus infection in an immunocompetent host. PMID- 11856245 TI - Gastrointestinal and respiratory Mycobacterium avium colonization and development of disseminated infection in HIV-positive patients. PMID- 11856246 TI - Infection on expeditions. PMID- 11856247 TI - Epidemiologic typing of international collections of Klebsiella spp.: computerized biochemical fingerprinting compared with serotyping, phage typing and pulsed-field gel electrophoresis. AB - OBJECTIVE: The Phene Plate (PhP) biochemical fingerprinting system is based on measurements of the kinetics of selected biochemical reactions performed in microtiter plates, and computerized data-processing. This study compared the performance of the PhP system as an epidemiologic tool with other commonly used typing systems. METHODS: PhP typing was applied to 107 nosocomial Klebsiella spp. isolates from 10 collections, mostly representing outbreaks. The results were compared with those obtained by capsular (K) serotyping, phage typing and, for a subset of isolates (n=33), pulsed-field gel electrophoresis (PFGE). RESULTS: Clusters of identical or closely related isolates based on serotype, phage type, PhP type and PFGE type were found in most collections. The typeability was 100%, 95%, 94% and 68% for PhP, K, PFGE and phage typing, respectively. The agreement between the typing methods was high (88-96%). The discriminatory power was high for PhP and PFGE (diversity index 0.95 and 0.97, respectively), but lower for phage typing (diversity index 0.91) and K typing (diversity index 0.87). CONCLUSIONS: Like serotyping and PFGE, PhP typing is useful in studies of the nosocomial epidemiology of Klebsiella spp. Combining PhP typing with PFGE or K typing rarely yielded additional information when comparing isolates within each collection, but PFGE sometimes discriminated between isolates of similar PhP type derived from different collections. PMID- 11856248 TI - Failure of antibiotic therapy in Staphylococcus epidermidis infection of implantable venous access devices in patients with AIDS, as documented by molecular typing. AB - OBJECTIVE: To study the efficacy and safety of high-concentration antibiotic locks in association with systemic antibiotherapy in Staphylococcus epidermidis infections of totally implantable venous access devices (TIVADs) in patients with AIDS. METHODS: Thirty-one episodes of S. epidermidis TIVAD infection were observed in nine patients. Locks consisted of high concentrations of aminoglycosides or vancomycin according to antibiogram results (susceptibility results obtained with the disk diffusion technique). Genotyping of bacterial strains was performed by pulsed-field gel electrophoresis (PFGE) in 26 of the 31 infectious episodes. RESULTS: The infections occurred within a median period of 62 catheter-days. The median duration of systemic antibiotic therapy was 17.5 days in association with a median of three antibiotic locks. Failure as defined by the occurrence of a novel episode within 2 months was observed in 17 of the 31 infections (58%). According to the PFGE results, relapse with the same strain of S. epidermidis or reinfection with a different strain of S. epidermidis could be assigned to 10 episodes which failed to respond to therapy. Relapse was observed in six of 10 episodes. Four episodes were followed by the occurrence of a novel infection with a different S. epidermidis strain. In one patient, a relapse occurred despite TIVAD removal. CONCLUSION: Our observations further document the poor efficacy of associating antibiotic locks with systemic antibiotic therapy for the treatment of TIVAD infections in patients with AIDS. PMID- 11856249 TI - The in vitro activity of moxifloxacin against Haemophilus influenzae and Moraxella catarrhalis explored using a pharmacodynamic model. AB - OBJECTIVE: To assess the antibacterial action of moxifloxacin on Haemophilus influenzae and Moraxella catarrhalis using an in vitro pharmacodynamic model of infection. METHODS: Serum concentrations in humans associated with doses of 400 mg once a day for 48 h were simulated and the antibacterial effect measured by the log change in viable count at intervals through the simulation compared to time zero and also the area under the bacterial kill curve (AUBKC). Wild-type strains of H. influenzae and M. catarrhalis with varying beta-lactam susceptibilities (moxifloxacin MICs /=1:32. The sensitivities of ELISA IgM for the first and the second serum samples compared with IIF were 42.8% and 46.6%, respectively, while the specificities were higher, i.e. 87% and 88.4%, respectively. The sensitivities of ELISA IgG for the first and the second samples were 42.8% and 53.3%, and the specificities were 77.9% and 76.8%, respectively. CONCLUSIONS: Although ELISA is simple to perform and easy to automate, we think that its advantages over indirect immunofluorescence and urinary antigen detection remain questionable. PMID- 11856251 TI - In vitro activity, postantibiotic effect and human monocyte activity of grepafloxacin against Legionella species. AB - OBJECTIVE: To determine the in vitro antimicrobial activity, postantibiotic effect (PAE) and human monocyte activity of grepafloxacin compared with sparfloxacin, ciprofloxacin, clarithromycin, erythromycin and rifampicin against 181 strains of Legionella pneumophila, nine strains of L. micdadei, 10 strains of L. dumoffii, seven strains of L. longbeachae and seven other Legionella strains. METHODS: MICs were determined by standard agar dilution using buffered yeast extract (BYE) agar. PAE and human monocyte activity were determined by standard culture techniques. RESULTS: Grepafloxacin, sparfloxacin and rifampicin were the most active agents against L. pneumophila (MIC90 10 days) neutropenia, such as patients undergoing bone marrow allografting or induction chemotherapy for acute leukemia. Initially, treatment was in the form of non-absorbable antibiotics, but this has been partially superseded by quinolone-containing regimens, in particular in the centers participating in EORTC trials. In the last two EORTC trials comparing different regimens for the treatment of febrile neutropenia, 57-73% of the patients were receiving GD. A French epidemiologic study, performed prospectively and consecutively in 36 oncohematology centers, has recently shown that 45% of febrile neutropenic patients receive digestive decontamination (DD) at the onset of their first febrile episode. The value of GD has been the subject of much controversy. Numerous trials, some of which were controlled, were performed in neutropenic patients in the 1980s, prior to trials of GD in intensive care units, but did not lead to a consensus in the medical community of the value of GD. Moreover, GD is not, or is infrequently, used in the USA. Apart from trials involving the quinolones, very few studies have been published during the last 10 years. Despite this, policies have not changed greatly in the various centers. The CLIOH group has gathered the opinions of experts invited to a multidisciplinary meeting that took place in Paris in October 1996. The text that follows summarizes the reflections arising from this forum. It should be noted that this meeting was not designed to be a consensus conference, but rather to re examine the correlation between the data in the literature and actual clinical practice and to highlight the main problems posed by DD in current oncohematology. The experts were separated into three working groups, each of which has drafted a report which appears in the text below. PMID- 11856289 TI - Molecular typing of fluoroquinolone-resistant and fluoroquinolone-susceptible Escherichia coli isolated from blood of neutropenic cancer patients in a single center. AB - OBJECTIVE: To investigate the molecular epidemiology of fluoroquinolone-resistant (FQ-R) and fluoroquinolone-susceptible (FQ-S) bacteremic Escherichia coli isolates from neutropenic patients by pulsed-field gel electrophoresis (PFGE) and random amplified polymorphic DNA (RAPD) analysis. METHODS: Nineteen FQ-R and 27 FQ-S isolates of E. coli, obtained from patients on a hematologic ward over a 7 year period, were genotyped by PFGE and RAPD using two different random primers (1247 and 1283). RESULTS: PFGE analysis was able to type all FQ-S isolates and most (17/19, 89%) FQ-R isolates of E. coli. All isolates were genotypically unrelated, with the exception of two indistinguishable FQ-R isolates from different patients in the same period. RAPD analysis typed all isolates, including those FQ-R isolates untypable by PFGE, but was unable to distinguish between some isolates that were different by PFGE. Using primer 1247, RAPD analysis identified six pairs and one triad, while primer 1283 identified seven pairs and one triad of indistinguishable isolates. CONCLUSIONS: No spread of epidemic FQ-R or FQ-S E. coli isolates was documented among neutropenic patients. RAPD analysis is a powerful genotyping method, but appeared to be less reproducible and discriminatory than PFGE for investigating E. coli isolates. PMID- 11856290 TI - A multicenter evaluation of genotypic methods for the epidemiologic typing of Legionella pneumophila serogroup 1: results of a pan-European study. AB - OBJECTIVES: To compare genotypic methods for epidemiologic typing of Legionella pneumophila serogroup (sg) 1, in order to determine the best available method within Europe for implementation and standardization by members of the European Working Group on Legionella Infections. METHODS: Coded isolates (114) of L. pneumophila sg 1 comprising one epidemiologically 'unrelated' (79) and one 'related' panel of isolates (35) were sent to 12 laboratories in 11 European countries. Analysis was undertaken in each laboratory using one or more of the following methods: ribotyping, restriction fragment length polymorphism analysis, restriction endonuclease analysis, pulsed-field gel electrophoresis (PFGE), PCR using arbitrary/repeat sequence primers (AP-, AP/rep-PCR), and amplified fragment length polymorphism (AFLP) analysis. Results were analyzed visually or using gel analysis software. Each method was assessed for its: index of discrimination (D), epidemiologic concordance (E), speed of application and ease of use. In addition, phenotypic analysis was performed in two laboratories using monoclonal antibodies (mAbs). RESULTS: The D of each of the genotypic methods ranged from 0.840 for ribotyping to 0.990 for PFGE using Sfil: E ranged from 0.06 for AP- and AP/rep PCR to 1.00 for ribotyping using Pstl/EcoRI and AFLP: in general, E was inversely related to D. Although offering only limited discrimination (D=0.838), mAb typing was both rapid and highly epidemiologically concordant (E=1.00). CONCLUSIONS: Two methods, PFGE using Sfil and AFLP, were selected for further study. AFLP is rapid and highly epidemiologically concordant (E=1.00), but is not highly discriminatory. This method will be developed as a rapid screening tool. PFGE using Sfil is highly discriminatory but, in the present study, yielded low values of E (0.12-0.71). Attempts will be made to rigorously standardize this method for use as the reference method. Primary screening of isolates by mAb subgrouping is recommended. PMID- 11856291 TI - Antimicrobial susceptibility of bacteria causing urinary tract infections in Latin American hospitals: results from the SENTRY Antimicrobial Surveillance Program (1997). AB - OBJECTIVE: To evaluate the antimicrobial susceptibility patterns among 469 pathogens isolated as a significant cause of urinary tract infections in 10 Latin American medical centers. METHODS: Consecutively collected isolates were susceptibility tested by broth microdilution methods, and selected isolates were characterized by molecular typing methods. RESULTS: Escherichia coli and Klebsiella spp. isolates revealed high rates of resistance to broad-spectrum penicillins and to fluoroquinolones. Ceftazidime MICs of >/=2 mg/L, suggesting the production of extended-spectrum b-lactamases (ESBLs), were observed in 37.7% of K. pneumoniae and 8.3% of Escherichia coli isolates. Enterobacter spp. isolates were characterized by high resistance rates to ciprofloxacin (35%) and to ceftazidime (45%), but they generally remained susceptible to cefepime (95% susceptible). Pseudomonas aeruginosa and Acinetobacter spp. were highly resistant to ciprofloxacin and ceftazidime. Imipenem was active against 80% of P. aeruginosa and 93% of Acinetobacter spp. isolates. CONCLUSIONS: Our results demonstrate a high level of resistance to various classes of antimicrobial agents among isolates causing nosocomial urinary tract infections in Latin American hospitals. Clonal dissemination of ESBL-producing K. pneumoniae strains was infrequent. PMID- 11856292 TI - In vitro activity of quinupristin/dalfopristin against selected bacterial pathogens isolated in Italy. AB - OBJECTIVE: To evaluate the activity of quinupristin/dalfopristin, a new injectable streptogramin, against 732 clinical strains recently isolated in Italy. METHODS: Susceptibility tests were performed according to NCCLS-guided MIC methodology. Pathogens included in the evaluation included 108 Staphylococcus aureus isolates, 124 coagulase-negative staphylococcal isolates, 158 Streptococcus pyogenes isolates, 30 Streptococcus agalactiae isolates, 30 b hemolytic streptococcal isolates, 18 Streptococcus sanguis isolates, 80 Streptococcus pneumoniae isolates, 69 Enterococcal isolates, 40 Haemophilus influenzae isolates, 30 Moraxella catarrhalis isolates and, finally, 30 Gram positive and 25 Gram-negative anaerobes. RESULTS: Quinupristin/dalfopristin inhibited Staphylococcus aureus and other Staphylococcus spp., irrespective of their oxacillin or erythromycin resistance phenotypes. Similarly, streptococci were fully inhibited by quinupristin/dalfopristin. Enterococcus faecalis was not included in the spectrum of this streptogramin, while isolates of Enterococcus faecium were inhibited by the new compound. Respiratory pathogens such as H. influenzae and M. catarrhalis were inhibited by quinupristin/dalfopristin as well as all Gram-negative anaerobes tested. CONCLUSIONS: These findings suggest a putative role for quinupristin/dalfopristin in the empirical treatment of severe nosocomial and community-acquired infections caused by pathogens often displaying resistance to multiple antibiotics. This drug may provide an alternative to glycopeptide compounds. PMID- 11856293 TI - Class I integrons: prevalence and impact on antibiotic susceptibility in 278 consecutive unrelated Gram-negative blood isolates. PMID- 11856294 TI - In vitro comparative activity of meropenem with 15 other antimicrobial agents against 1798 Pseudomonas aeruginosa isolates in a French multicenter study. PMID- 11856295 TI - First report of vancomycin-resistant Enterococcus faecium isolated in Poland. PMID- 11856296 TI - Is routinely searching for halophilic vibrios useful in non-coastal areas? PMID- 11856297 TI - Empirical antibiotic therapy: assessment of agreement between pediatricians. PMID- 11856298 TI - Plant alpha-amylase inhibitors and their interaction with insect alpha-amylases. AB - Insect pests and pathogens (fungi, bacteria and viruses) are responsible for severe crop losses. Insects feed directly on the plant tissues, while the pathogens lead to damage or death of the plant. Plants have evolved a certain degree of resistance through the production of defence compounds, which may be aproteic, e.g. antibiotics, alkaloids, terpenes, cyanogenic glucosides or proteic, e.g. chitinases, beta-1,3-glucanases, lectins, arcelins, vicilins, systemins and enzyme inhibitors. The enzyme inhibitors impede digestion through their action on insect gut digestive alpha-amylases and proteinases, which play a key role in the digestion of plant starch and proteins. The natural defences of crop plants may be improved through the use of transgenic technology. Current research in the area focuses particularly on weevils as these are highly dependent on starch for their energy supply. Six different alpha-amylase inhibitor classes, lectin-like, knottin-like, cereal-type, Kunitz-like, gamma purothionin-like and thaumatin-like could be used in pest control. These classes of inhibitors show remarkable structural variety leading to different modes of inhibition and different specificity profiles against diverse alpha-amylases. Specificity of inhibition is an important issue as the introduced inhibitor must not adversely affect the plant's own alpha-amylases, nor the nutritional value of the crop. Of particular interest are some bifunctional inhibitors with additional favourable properties, such as proteinase inhibitory activity or chitinase activity. The area has benefited from the recent determination of many structures of alpha-amylases, inhibitors and complexes. These structures highlight the remarkable variety in structural modes of alpha-amylase inhibition. The continuing discovery of new classes of alpha-amylase inhibitor ensures that exciting discoveries remain to be made. In this review, we summarize existing knowledge of insect alpha-amylases, plant alpha-amylase inhibitors and their interaction. Positive results recently obtained for transgenic plants and future prospects in the area are reviewed. PMID- 11856299 TI - Dimerization of mammalian adenylate cyclases. AB - Mammalian adenylate cyclases are predicted to possess complex topologies, comprising two cassettes of six transmembrane-spanning motifs followed by a cytosolic, catalytic ATP-binding domain. Recent studies have begun to provide insights on the tertiary assembly of these proteins; crystallographic analysis has revealed that the two cytosolic domains dimerize to form a catalytic core, while more recent biochemical and cell biological analysis shows that the two transmembrane cassettes also associate to facilitate the functional assembly and trafficking of the enzyme. The older literature had suggested that adenylate cyclases might form higher order aggregates, although the methods used did not necessarily provide convincing evidence of biologically relevant events. In the present study, we have pursued this question by a variety of approaches, including rescue or suppression of function by variously modified molecules, coimmunoprecipitation and fluorescence resonance energy transfer (FRET) analysis between molecules in living cells. The results strongly suggest that adenylate cyclases dimerize (or oligomerize) via their hydrophobic domains. It is speculated that this divalent property may allow adenylate cyclases to participate in multimeric signaling assemblies. PMID- 11856300 TI - The structure of the carbohydrate backbone of the lipopolysaccharide from Acinetobacter baumannii strain ATCC 19606. AB - The chemical structure of the phosphorylated carbohydrate backbone of the lipopolysaccharide (LPS) from Acinetobacter baumannii strain ATCC 19606 was investigated by chemical analysis and NMR spectroscopy of oligosaccharides obtained after deacylation or mild acid hydrolysis. From the combined information the following carbohydrate backbones can be deduced: where R1 = H and R2 = alpha Glcp-(1-->2)-beta-Glcp-(1-->4)-beta-Glcp-(1-->4)-beta-Glcp-(1 as major and R1 = Ac and R2 = H as minor products. All monosaccharides are d-configured. Also, smaller oligosaccharide phosphates were identified that are thought to represent degradation products of the above structures. PMID- 11856301 TI - Domain IV of mouse laminin beta1 and beta2 chains. AB - Domain IV, consisting of about 230 residues, represents a particular protein module so far found only in laminin beta1 and beta2 chains. Both domains were obtained by recombinant production in mammalian cells. They showed a globular structure, as expected from electron microscopic examination of laminins. Fragment beta1IV was obtained as a monomer and a disulfide-bonded dimer, and both were modified to approximately 50% by a single chondroitin sulfate chain attached to Ser721 of an SGD consensus sequence. Dimerization is caused by an odd number of cysteines, with three of them having a partial thiol character. Whether both modifications also occur in tissue forms of laminin remains to be established. Fragment beta2IV was only obtained as a monomer, as it lacked one crucial cysteine and the SGD sequence. It required, however, the presence of two adjacent LE modules for proper folding. Polyclonal antibodies raised against both fragments showed no cross-reaction with each other and allowed establishment of beta chain-specific radioimmunoassays and light and electron microscopic immunostaining of tissues. This demonstrated a 5-25-fold lower content of beta2 compared with beta1 chains in various tissue extracts of adult mice. Tissues derived from beta2-deficient mice failed to react with the beta2-specific antibodies but showed a twofold higher content of beta1 than heterozygotes. The antibodies to beta2 showed broader tissue staining than reported previously, including in particular a distinct reaction with the extrasynaptic endomysium of skeletal muscle. Immunogold staining localized both beta chains primarily to basement membranes of kidney, muscle and various other tissues. PMID- 11856302 TI - Structure of peptidase T from Salmonella typhimurium. AB - The structure of peptidase T, or tripeptidase, was determined by multiple wavelength anomalous dispersion (MAD) methodology and refined to 2.4 A resolution. Peptidase T comprises two domains; a catalytic domain with an active site containing two metal ions, and a smaller domain formed through a long insertion into the catalytic domain. The two metal ions, presumably zinc, are separated by 3.3 A, and are coordinated by five carboxylate and histidine ligands. The molecular surface of the active site is negatively charged. Peptidase T has the same basic fold as carboxypeptidase G2. When the structures of the two enzymes are superimposed, a number of homologous residues, not evident from the sequence alone, could be identified. Comparison of the active sites of peptidase T, carboxypeptidase G2, Aeromonas proteolytica aminopeptidase, carboxypeptidase A and leucine aminopeptidase reveals a common structural framework with interesting similarities and differences in the active sites and in the zinc coordination. A putative binding site for the C-terminal end of the tripeptide substrate was found at a peptidase T specific fingerprint sequence motif. PMID- 11856303 TI - The unique sites in SulA protein preferentially cleaved by ATP-dependent Lon protease from Escherichia coli. AB - SulA protein is known to be one of the physiological substrates of Lon protease, an ATP-dependent protease from Escherichia coli. In this study, we investigated the cleavage specificity of Lon protease toward SulA protein. The enzyme was shown to cleave approximately 27 peptide bonds in the presence of ATP. Among them, six peptide bonds were cleaved preferentially in the early stage of digestion, which represented an apparently unique cleavage sites with mainly Leu and Ser residues at the P1, and P1' positions, respectively, and one or two Gln residues in positions P2-P5. They were located in the central region and partly in the C-terminal region, both of which are known to be important for the function of SulA, such as inhibition of cell growth and interaction with Lon protease, respectively. The other cleavage sites did not represent such consensus sequences, though hydrophobic or noncharged residues appeared to be relatively preferred at the P1 sites. On the other hand, the cleavage in the absence of ATP was very much slower, especially in the central region, than in the presence of ATP. The central region was predicted to be rich in alpha helix and beta sheet structures, suggesting that the enzyme required ATP for disrupting such structures prior to cleavage. Taken together, SulA is thought to contain such unique cleavage sites in its functionally and structurally important regions whose preferential cleavage accelerates the ATP-dependent degradation of the protein by Lon protease. PMID- 11856304 TI - Identification and properties of type I-signal peptidases of Bacillus amyloliquefaciens. AB - The use of Bacillus amyloliquefaciens for enzyme production and its exceptional high protein export capacity initiated this study where the presence and function of multiple type I signal peptidase isoforms was investigated. In addition to type I signal peptidases SipS(ba) [Meijer, W.J.J., de Jong, A., Bea, G., Wisman, A., Tjalsma, H., Venema, G., Bron, S. & van Dijl, J.M. (1995) Mol. Microbiol. 17, 621-631] and SipT(ba) [Hoang, V. & Hofemeister, J. (1995) Biochim. Biophys. Acta 1269, 64-68] which were previously identified, here we present evidence for two other Sip-like genes in B. amyloliquefaciens. Same map positions as well as sequence motifs verified that these genes encode homologues of Bacillus subtilis SipV and SipW. SipU-encoding DNA was not found in B. amyloliquefaciens. SipW encoding DNA was also found for other Bacillus strains representing different phylogenetic groups, but not for Bacillus stearothermophilus and Thermoactinomyces vulgaris. The absence of these genes, however, could have been overlooked due to sequence diversity. Sequence alignments of 23 known Sip-like proteins from Bacillus origin indicated further branching of the P-group signal peptidases into clusters represented by B. subtilis SipV, SipS-SipT-SipU and B. anthracis Sip3-Sip5 proteins, respectively. Each B. amyloliquefaciens sip(ba) gene was expressed in an Escherichia coli LepBts mutant and tested for genetic complementation of the temperature sensitive (TS) phenotype as well as pre-OmpA processing. Although SipS(ba) as well as SipT(ba) efficiently restored processing of pre-OmpA in E. coli, only SipS(ba) supported growth at TS conditions, indicating functional diversity. Changed properties of the sip(ba) gene disruption mutants, including cell autolysis, motility, sporulation, and nuclease activities, seemed to correlate with specificities and/or localization of B. amyloliquefaciens SipS, SipT and SipV isoforms. PMID- 11856305 TI - Coordination chemistry of iron(III)-porphyrin-antibody complexes. AB - An artificial peroxidase-like hemoprotein has been obtained by associating a monoclonal antibody, 13G10, and its iron(III)-alpha,alpha,alpha,beta-meso tetrakis(ortho-carboxyphenyl)porphyrin [Fe(ToCPP)] hapten. In this antibody, about two-thirds of the porphyrin moiety is inserted in the binding site, its ortho-COOH substituents being recognized by amino-acids of the protein, and a carboxylic acid side chain of the protein acts as a general acid base catalyst in the heterolytic cleavage of the O-O bond of H2O2, but no amino-acid residue is acting as an axial ligand of the iron. We here show that the iron of 13G10 Fe(ToCPP) is able to bind, like that of free Fe(ToCPP), two small ligands such as CN-, but only one imidazole ligand, in contrast to to the iron(III) of Fe(ToCPP) that binds two. This phenomenon is general for a series of monosubstituted imidazoles, the 2- and 4-alkyl-substituted imidazoles being the best ligands, in agreement with the hydrophobic character of the antibody binding site. Complexes of antibody 13G10 with less hindered iron(III)-tetraarylporphyrins bearing only one [Fe(MoCPP)] or two meso-[ortho-carboxyphenyl] substituents [Fe(DoCPP)] also bind only one imidazole. Finally, peroxidase activity studies show that imidazole inhibits the peroxidase activity of 13G10-Fe(ToCPP) whereas it increases that of 13G10-Fe(DoCPP). This could be interpreted by the binding of the imidazole ligand on the iron atom which probably occurs in the case of 13G10-Fe(ToCPP) on the less hindered face of the porphyrin, close to the catalytic COOH residue, whereas in the case of 13G10-Fe(DoCPP) it can occur on the other face of the porphyrin. The 13G10-Fe(DoCPP)-imidazole complex thus constitutes a nice artificial peroxidase like hemoprotein, with the axial imidazole ligand of the iron mimicking the proximal histidine of peroxidases and a COOH side chain of the antibody acting as a general acid-base catalyst like the distal histidine of peroxidases does. PMID- 11856306 TI - Characterization of glycosphingolipids from Schistosoma mansoni eggs carrying Fuc(alpha1-3)GalNAc-, GalNAc(beta1-4)[Fuc(alpha1-3)]GlcNAc- and Gal(beta1 4)[Fuc(alpha1-3)]GlcNAc- (Lewis X) terminal structures. AB - The carbohydrate moieties of glycosphingolipids from eggs of the human parasite, Schistosoma mansoni, were enzymatically released, labelled with 2-aminopyridine (PA), fractionated and analysed by linkage analysis, partial hydrolysis, enzymatic cleavage, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and nano-electrospray ionization mass spectrometry. Apart from large, highly fucosylated structures with five to seven HexNAc residues, we found short, oligofucosylated species containing three to four HexNAc residues. Their structures have been determined as Fuc(alpha1-3)GalNAc(beta1-4)[ +/- Fuc (alpha1 3)]GlcNAc(beta1-3)GalNAc(beta1-4)Glc-PA, GalNAc(beta1-4)[Fuc(alpha1 3)]GlcNAc(beta1-3)GlcNAc(beta1-3)GalNAc(beta1-4) Glc-PA, Fuc(alpha1 3)GalNAc(beta1-4)[Fuc(alpha1-3)]GlcNAc(beta1-4) GlcNAc(beta1-3)GalNAc(beta1-4)Glc PA, and Fuc(alpha1-3) GalNAc(beta1-4)[ +/- Fuc(alpha1-2) +/- Fuc(alpha1 2)Fuc(alpha1-3)]Glc NAc(beta1-3)GlcNAc(beta1-3)GalNAc(beta1-4)Glc-PA. The last structure exhibits a trifucosyl sidechain previously identified on the cercarial glycocalyx. These structures stress the importance of 3-fucosylated GalNAc as a terminal epitope in schistosome glycoconjugates. To what degree these glycans contribute to the pronounced antigenicity of S. mansoni egg glycolipids remains to be determined. In addition, we have identified the compounds GlcNAc(beta1 3)GalNAc(beta1-4)Glc-PA, Gal(beta1-4)[Fuc(alpha1-3)]GlcNAc(beta1-3) GalNAc (beta1 4)Glc-PA, the latter of which is a Lewis X-pentasaccharide identical to that present on cercarial glycolipids, as well as Gal(beta1-3)GalNAc(1-4)Gal(1-4)Glc PA, which corresponds to asialogangliotetraosylceramide and is most probably derived from the mammalian host. PMID- 11856307 TI - Different mechanisms for cellular internalization of the HIV-1 Tat-derived cell penetrating peptide and recombinant proteins fused to Tat. AB - Translocation through the plasma membrane is a major limiting step for the cellular delivery of macromolecules. A promising strategy to overcome this problem consists in the chemical conjugation (or fusion) to cell penetrating peptides (CPP) derived from proteins able to cross the plasma membrane. A large number of different cargo molecules such as oligonucleotides, peptides, peptide nucleic acids, proteins or even nanoparticles have been internalized in cells by this strategy. One of these translocating peptides was derived from the HIV-1 Tat protein. The mechanisms by which CPP enter cells remain unknown. Recently, convincing biochemical and genetic findings has established that the full-length Tat protein was internalized in cells via the ubiquitous heparan sulfate (HS) proteoglycans. We demonstrate here that the short Tat CPP is taken up by a route that does not involve the HS proteoglycans. PMID- 11856308 TI - Cell surface heparan sulfate proteoglycans: target and partners of the basic fibroblast growth factor in rat Sertoli cells. AB - Basic fibroblast growth factor (bFGF) regulates diversified biological functions in rat Sertoli cells. This report demonstrates that bFGF inhibits steroidogenesis in developing rat Sertoli cells. Follicle stimulating hormone (FSH)-stimulated estradiol production was reduced by bFGF. Moreover, the amount of cytochrome P450 aromatase, responsible for the irreversible transformation of androgens into estrogens, is decreased by bFGF at the transcriptional level. The bFGF inhibitory effect was also observed in the presence of dibutyryl-cAMP, cholera toxin or RO 20-1724, all inducing high levels of cAMP, the second messenger of FSH. Heparan sulfate proteoglycans (HSPGs) were shown to be required as cofactors for bFGF signaling. Indeed, sodium chlorate, described to drastically decrease proteoglycan sulfation, abolishes the bFGF downregulation of FSH-stimulated estradiol synthesis previously observed. Glypican-1, syndecan-1 and -4, potential bFGF coreceptors, are mainly regulated at the transcriptional level. This report shows that the bFGF regulation of their expression specifically depends on the nature of HSPG and of the Sertoli cell developmental stage. In conclusion, HSPG are partners and the target of bFGF in rat Sertoli cells. PMID- 11856309 TI - Antibacterial peptides in stimulated human granulocytes: characterization of ubiquitinated histone H1A. AB - Antibacterial peptides were isolated from human peripheral granulocytes of a healthy donor who had been treated with granulocyte-colony stimulating factor (G CSF) and cortisol. Peptides were solubilized in acidified chloroform/methanol, and partitioned in chloroform/methanol/water. Water- soluble polypeptides were separated by cation-exchange and reversed-phase chromatography. Several previously characterized antibacterial polypeptides were identified; defensins 1 3, defensin 4, lysozyme, eosinophil cationic protein, and calgranulin A. In addition, several histone fragments were isolated and exhibited activity against the Gram- positive bacterium Bacillus megaterium strain Bm11. These fragments included two C-terminal fragments of histone H1A, three C-terminal fragments of histone H1D, one fragment of histone H1B, and two fragments of histone H4. The molecular masses of both histone H1A fragments, as determined by electrospray (ES) MS, were 270 Da higher than those calculated from their amino acid sequences. The two histone H1A fragments corresponded to Lys152-Lys222 (7527 +/- 1 Da) and Lys167-Lys222 (6023 +/- 1 Da). Tandem MS (MS/MS) of the 7.5 kDa and 6.0 kDa fragments indicated that the post-translational modification is on Lys222, the epsilon-amino group of which was conjugated with the alpha-carboxyl group of the tripeptide Arg-Gly-Gly. This finding was substantiated by digestion of the 7.5-kDa polypeptide with trypsin and analysis of the resulting peptides by ES MS and MS/MS. The tripeptide Arg-Gly-Gly corresponded uniquely to the three C terminal residues of ubiquitin, demonstrating the presence of ubiquitinated histone H1A. PMID- 11856310 TI - Biosynthesis of riboflavin: 6,7-dimethyl-8-ribityllumazine synthase of Schizosaccharomyces pombe. AB - A cDNA sequence from Schizosaccharomyces pombe with similarity to 6,7-dimethyl-8 ribityllumazine synthase was expressed in a recombinant Escherichia coli strain. The recombinant protein is a homopentamer of 17-kDa subunits with an apparent molecular mass of 87 kDa as determined by sedimentation equilibrium centrifugation (it sediments at an apparent velocity of 5.0 S at 20 degrees C). The protein has been crystallized in space group C2221. The crystals diffract to a resolution of 2.4 A. The enzyme catalyses the formation of 6,7-dimethyl-8 ribityllumazine from 5-amino-6-ribitylamino-2,4(1H,3H)-pyrimidinedione and 3,4 dihydroxy- 2-butanone 4-phosphate. Steady-state kinetic analysis afforded a vmax value of 13 000 nmol.mg-1.h-1 and Km values of 5 and 67 microm for 5-amino-6 ribitylamino-2,4(1H,3H)-pyrimidinedione and 3,4-dihydroxy-2-butanone 4-phosphate, respectively. The enzyme binds riboflavin with a Kd of 1.2 microm. The fluorescence quantum yield of enzyme-bound riboflavin is < 2% as compared with that of free riboflavin. The protein/riboflavin complex displays an optical transition centered around 530 nm as shown by absorbance and CD spectrometry which may indicate a charge transfer complex. Replacement of tryptophan 27 by tyrosine or phenylalanine had only minor effects on the kinetic properties, but complexes of the mutant proteins did not show the anomalous long wavelength absorbance of the wild-type protein. The replacement of tryptophan 27 by aliphatic amino acids substantially reduced the affinity of the enzyme for riboflavin and for the substrate, 5-amino-6-ribitylamino-2,4(1H,3H) pyrimidinedione. PMID- 11856311 TI - Comparative structure analysis of proteinase inhibitors from the desert locust, Schistocerca gregaria. AB - The solution structure of three small serine proteinase inhibitors, two natural and one engineered protein, SGCI (Schistocerca gregaria chymotrypsin inhibitor), SGCI[L30R, K31M] and SGTI (Schistocerca gregaria trypsin inhibitor), were determined by homonuclear NMR-spectroscopy. The molecules exhibit different specificities towards target proteinases, where SGCI is a good chymotrypsin inhibitor, its mutant is a potent trypsin inhibitor, and SGTI inhibits both proteinases weakly. Interestingly, SGTI is a much better inhibitor of insect proteinases than of the mammalian ones used in common assays. All three molecules have a similar fold composed from three antiparallel beta-pleated sheets with three disulfide bridges. The proteinase binding loop has a somewhat distinct geometry in all three peptides. Moreover, the stabilization of the structure is different in SGCI and SGTI. Proton-deuterium exchange experiments are indicative of a highly rigid core in SGTI but not in SGCI. We suggest that the observed structural properties play a significant role in the specificity of these inhibitors. PMID- 11856312 TI - NBR1 interacts with fasciculation and elongation protein zeta-1 (FEZ1) and calcium and integrin binding protein (CIB) and shows developmentally restricted expression in the neural tube. AB - NBR1 (named as next to BRCA1) was originally cloned as a candidate gene for the ovarian cancer antigen CA125, using expression cloning with the anti-CA125 Ig, OC125. NBR1 has been of interest due to its position close to BRCA1, although no involvement in breast or ovarian cancer has been demonstrated. Recently, the antigen CA125 has been cloned, and identified as a new mucin, MUC16, entirely different from NBR1. The function of NBR1 remains unknown. To investigate its function, a yeast two-hybrid study was performed to identify interacting protein partners that may reflect a biological role for this protein. Here, we show that NBR1 interacts with two proteins; fasciculation and elongation protein zeta-1 (FEZ1), a PKCzeta interacting protein, and calcium and integrin binding protein (CIB), which is associated with polo-like kinases Fnk/Snk and the Alzheimer's disease presenilin 2 protein. Co-transfection of FEZ1 and NBR1 showed overlapping localization in the cytoplasm, whereas coexpression of NBR1 and CIB resulted in a shift of CIB protein expression from the nucleus to the perinuclear compartment. FEZ1 is highly expressed in the brain and in situ hybridization analysis of Nbr1 showed that its expression is also regulated in the murine brain during development. These data suggest that NBR1 may function, through interaction with CIB and FEZ1 in cell signalling pathways, with a developmentally restricted expression suggesting a possible role in neural development. PMID- 11856314 TI - Evaluation of potential regulatory elements identified as DNase I hypersensitive sites in the CFTR gene. AB - The cystic fibrosis transmembrane conductance regulator (CFTR) gene shows a complex pattern of expression, with temporal and spatial regulation that is not accounted for by elements in the promoter. One approach to identifying the regulatory elements for CFTR is the mapping of DNase I hypersensitive sites (DHS) within the locus. We previously identified at least 12 clusters of DHS across the CFTR gene and here further evaluate DHS in introns 2, 3, 10, 16, 17a, 18, 20 and 21 to assess their functional importance in regulation of CFTR gene expression. Transient transfections of enhan- cer/reporter constructs containing the DHS regions showed that those in introns 20 and 21 augmented the activity of the CFTR promoter. Structural analysis of the DNA sequence at the DHS suggested that only the one intron 21 might be caused by inherent DNA structures. Cell specificity of the DHS suggested a role for the DHS in introns 2 and 18 in CFTR expression in some pancreatic duct cells. Finally, regulatory elements at the DHS in introns 10 and 18 may contribute to upregulation of CFTR gene transcription by forskolin and mitomycin C, respectively. These data support a model of regulation of expression of the CFTR gene in which multiple elements contribute to tightly co-ordinated expression in vivo. PMID- 11856313 TI - Two conserved domains in regulatory B subunits mediate binding to the A subunit of protein phosphatase 2A. AB - Protein phosphatase 2A (PP2A) is an abundant heterotrimeric serine/threonine phosphatase containing highly conserved structural (A) and catalytic (C) subunits. Its diverse functions in the cell are determined by its association with a highly variable regulatory and targeting B subunit. At least three distinct gene families encoding B subunits are known: B/B55/CDC55, B'/B56/RTS1 and B"/PR72/130. No homology has been identified among the B families, and little is known about how these B subunits interact with the PP2A A and C subunits. In vitro expression of a series of B56alpha fragments identified two distinct domains that bound independently to the A subunit. Sequence alignment of these A subunit binding domains (ASBD) identified conserved residues in B/B55 and PR72 family members. The alignment successfully predicted domains in B55 and PR72 subunits that similarly bound to the PP2A A subunit. These results suggest that these B subunits share a common core structure and mode of interaction with the PP2A holoenzyme. PMID- 11856315 TI - N-Methylation in polylegionaminic acid is associated with the phase-variable epitope of Legionella pneumophila serogroup 1 lipopolysaccharide. Identification of 5-(N,N-dimethylacetimidoyl)amino and 5-acetimidoyl(N-methyl)amino-7-acetamido 3,5,7,9-tetradeoxynon-2-ulosonic acid in the O-chain polysaccharide. AB - Previously, a phase-variable epitope was detected in the virulent wild-type strain RC1 of Legionella pneumophila serogroup 1 subgroup OLDA using a lipopolysaccharide-specific monoclonal antibody, mAb 2625 [Luneberg, E., Zahringer, U., Knirel, Y. A., Steinmann, D., Hartmann, M., Steinmetz, I., Rohde, M., Kohl, J. & Frosch, M. (1998) J.Exp. Med. 188, 49-60]. In the present study, an isogenic mutant strain, termed 5215, was constructed by deletion of genes involved in the biosynthesis of the mAb 2625 epitope. Mutant 5215 was as virulent as the parental wild-type RC1 but did not bind mAb 2625. The two strains showed no difference in the core oligosaccharide and lipid A but in the O-chain polysaccharide structure, which is a homopolymer of 5-acetimidoylamino-7 acetamido-3,5,7,9-tetradeoxy-d-glycero-d-galacto-non-2-ulosonic acid (a derivative of legionaminic acid). NMR spectroscopic studies revealed a hitherto unknown modification of bacterial polysaccharides in the wild-type strain, namely N-methylation of the 5-acetimidoylamino group on a single legionaminic acid residue that is located, most likely, proximal to the core oligosaccharide. Two major N-methylated substituents, the (N,N-dimethylacetimidoyl)amino and acetimidoyl(N-methyl) amino groups, could be allocated to the long- and middle chain O-polysaccharide species, respectively. N-Methylation of legionaminic acid that was absent from the isogenic mutant 5215 and from the spontaneous phase variant 811, correlated with the presence of the mAb 2625 epitope. PMID- 11856316 TI - Epitope mapping of the O-chain polysaccharide of Legionella pneumophila serogroup 1 lipopolysaccharide by saturation-transfer-difference NMR spectroscopy. AB - Two modifications of 5-acetimidoylamino-7-acetamido-3,5,7,9-tetradeoxy-D-glycero D-galacto-non-2-ulosonic acid (5-N-acetimidoyl-7-N-acetyllegionaminic acid) in the O-chain polysaccharide (OPS) of the Legionella pneumophila serogroup 1 lipopolysaccharide (LPS) concern N-methylation of the 5-N-acetimidoyl group in legionaminic acid. Both N-methylated substituents, the (N,N-dimethylacetimidoyl) amino and acetimidoyl(N-methyl)amino group, could be allocated to one single legionaminic acid residue in the long- and middle-chain OPS, respectively. Using mutants devoid of N-methylated legionaminic acid derivatives, it could be shown that N-methylation of legionaminic acid correlated with the expression of the mAb 2625 epitope. In the present study we investigated the binding of the LPS specific monoclonal antibody mAb 2625 to isolated OPS with surface-plasmon resonance biomolecular interaction analysis and saturation-transfer-difference (STD) NMR spectroscopy in order to map the mAb 2625 epitope on a molecular level. It could be demonstrated that the binding affinity of the N-methylated legionaminic acid derivatives was independent from the size of the isolated OPS molecular species. In addition, STD NMR spectroscopic studies with polysaccharide ligands with an average molecular mass of up to 14 kDa revealed that binding was mainly mediated via the N-methylated acetimidoylamino group and via the closely located 7-N-acetyl group of the respective legionaminic acid residue, thus indicating these derivatives to represent the major epitope of mAb 2625. PMID- 11856317 TI - The involvement of human ribonucleases H1 and H2 in the variation of response of cells to antisense phosphorothioate oligonucleotides. AB - We have analyzed the response of a number of human cell lines to treatment with antisense oligodeoxynucleotides (ODNs) directed against RNA polymerase II, replication protein A, and Ha-ras. ODN-delivery to the cells was liposome mediated or via electroporation, which resulted in different intracellular locations of the ODNs. The ODN-mediated target mRNA reduction varied considerably between the cell lines. In view of the essential role of RNase H activity in this response, RNase H was analyzed. The mRNA levels of RNase H1 and RNase H2 varied considerably in the cell lines examined in this study. The intracellular localization of the enzymes, assayed by green-fluorescent protein fusions, showed that RNase H1 was present throughout the whole cell for all cell types analyzed, whereas RNase H2 was restricted to the nucleus in all cells except the prostate cancer line 15PC3 that expressed the protein throughout the cell. Whole cell extracts of the cell lines yielded similar RNase H cleavage activity in an in vitro liquid assay, in contrast to the efficacy of the ODNs in vivo. Overexpression of RNase H2 did not affect the response to ODNs in vivo. Our data imply that in vivo RNase H activity is not only due to the activity assayed in vitro, but also to an intrinsic property of the cells. RNase H1 is not likely to be a major player in the antisense ODN-mediated degradation of target mRNAs. RNase H2 is involved in the activity assayed in vitro. The presence of cell-type specific factors affecting the activity and localization of RNase H2 is strongly suggested. PMID- 11856318 TI - Functional expression of Pseudomonas aeruginosa GDP-4-keto-6-deoxy-D-mannose reductase which synthesizes GDP-rhamnose. AB - Pseudomonas aeruginosa is an opportunistic Gram-negative bacterium that causes severe infections in a number of hosts from plants to mammals. A-band lipopolysaccharide of P. aeruginosa contains d-rhamnosylated O-antigen. The synthesis of GDP-D-rhamnose, the d-rhamnose donor in d-rhamnosylation, starts from GDP-D-mannose. It is first converted by the GDP-mannose-4,6-dehydratase (GMD) into GDP-4-keto-6-deoxy-D-mannose, and then reduced to GDP-D-rhamnose by GDP-4-keto-6-deoxy-D-mannose reductase (RMD). Here, we describe the enzymatic characterization of P. aeruginosa RMD expressed in Saccharomyces cerevisiae. Previous success in functional expression of bacterial gmd genes in S. cerevisiae allowed us to convert GDP-D-mannose into GDP-4-keto-6-deoxy-D-mannose. Thus, coexpression of the Helicobacter pylori gmd and P. aeruginosa rmd genes resulted in conversion of the 4-keto-6-deoxy intermediate into GDP-deoxyhexose. This synthesized GDP-deoxyhexose was confirmed to be GDP-rhamnose by HPLC, matrix assisted laser desorption/ionization time-of-flight MS, and finally NMR spectroscopy. The functional expression of P. aeruginosa RMD in S. cerevisiae will provide a tool for generating GDP-rhamnose for in vitro rhamnosylation of glycoprotein and glycopeptides. PMID- 11856319 TI - Expression of the Pycnoporus cinnabarinus laccase gene in Aspergillus niger and characterization of the recombinant enzyme. AB - Pycnoporus cinnabarinus laccase lac1 gene was overexpressed in Aspergillus niger, a well-known fungal host producing a large amount of homologous or heterologous enzymes for industrial applications. The corresponding cDNA was placed under the control of the glyceraldehyde-3-phosphate dehydrogenase promoter as a strong and constitutive promoter. The laccase signal peptide or the glucoamylase preprosequence of A. niger was used to target the secretion. Both signal peptides directed the secretion of laccase into the culture medium as an active protein, but the A. niger preprosequence allowed an 80-fold increase in laccase production. The identity of the recombinant protein was further confirmed by immunodetection using Western blot analysis and N-terminal sequencing. The molecular mass of the mature laccase was 70 kDa as expected, similar to that of the native form, suggesting no hyperglycosylation. The recombinant laccase was purified in a three-step procedure including a fractionated precipitation using ammonium sulfate, and a concentration by ultrafiltration followed by a Mono Q column. All the characteristics of the recombinant laccase are in agreement with those of the native laccase. This is the first report of the production of a white-rot laccase in A. niger. PMID- 11856320 TI - Absence of the psbH gene product destabilizes photosystem II complex and bicarbonate binding on its acceptor side in Synechocystis PCC 6803. AB - The PsbH protein, a small subunit of the photosystem II complex (PSII), was identified as a 6-kDa protein band in the PSII core and subcore (CP47-D1-D2-cyt b 559) from the wild-type strain of the cyanobacterium Synechocystis PCC 6803. The protein was missing in the D1-D2-cytochrome b-559 complex and also in all PSII complexes isolated from IC7, a mutant lacking the psbH gene. The following properties of PSII in the mutant contrasted with those in wild-type: (a) CP47 was released during nondenaturing electrophoresis of the PSII core isolated from IC7; (b) depletion of CO2 resulted in a reversible decrease of the QA- reoxidation rate in the IC7 cells; (c) light-induced decrease in PSII activity, measured as 2,5-dimethyl-benzoquinone-supported Hill reaction, was strongly dependent on the HCO3- concentration in the IC7 cells; and (d) illumination of the IC7 cells lead to an extensive oxidation, fragmentation and cross-linking of the D1 protein. We did not find any evidence for phosphorylation of the PsbH protein in the wild type strain. The results showed that in the PSII complex of Synechocystis attachment of CP47 to the D1-D2 heterodimer appears weakened and binding of bicarbonate on the PSII acceptor side is destabilized in the absence of the PsbH protein. PMID- 11856321 TI - Mutational analysis of the C-domain in nonribosomal peptide synthesis. AB - The initial condensation event in the nonribosomal biosynthesis of the peptide antibiotics gramicidin S and tyrocidine A takes place between a phenylalanine activating racemase GrsA/TycA and the first proline-activating module of GrsB/TycB. Recently we established a minimal in vitro model system for NRPS with recombinant His6-tagged GrsA (GrsAPhe-ATE; 127 kDa) and TycB1 (TycB1Pro-CAT; 120 kDa) and demonstrated the catalytic function of the C-domain in TycB1Pro-CAT to form a peptide bond between phenylalanine and proline during diketopiperazine formation (DKP). In this work we took advantage of this system to identify catalytically important residues in the C-domain of TycB1Pro-CAT using site directed mutagenesis and peptide mapping. Mutations in TycB1Pro-CAT of 10 strictly conserved residues among 80 other C-domains with potential catalytic function, revealed that only R62A, H147R and D151N are impaired in peptide-bond formation. All other mutations led to either unaffected (Q19A, C154A/S, Y166F/W and R284A) or insoluble proteins (H146A, R67A and W202L). Although 100 nm of the serine protease inhibitors N-alpha-tosyl-l-phenylalanylchloromethane or phenylmethanesulfonyl fluoride completely abolished DKP synthesis, no covalently bound inhibitor derivatives in the C-domain could be identified by peptide mapping using HPLC-MS. Though the results do not reveal a particular mechanism for the C-domain, they exhibit a possible way of catalysis analogous to the functionally related enzymes chloramphenicol acetyltransferase and dihydrolipoyl transacetylase. Based on this, we propose a mechanism in which one catalytic residue (H147) and two other structural residues (R62 and D151) are involved in amino-acid condensation. PMID- 11856322 TI - Engineering of a monomeric and low-glycosylated form of human butyrylcholinesterase: expression, purification, characterization and crystallization. AB - Human butyrylcholinesterase (BChE; EC 3.1.1.8) is of particular interest because it hydrolyzes or scavenges a wide range of toxic compounds including cocaine, organophosphorus pesticides and nerve agents. The relative contribution of each N linked glycan for the solubility, the stability and the secretion of the enzyme was investigated. A recombinant monomeric BChE lacking four out of nine N glycosylation sites and the C-terminal oligomerization domain was stably expressed as a monomer in CHO cells. The purified recombinant BChE showed catalytic properties similar to those of the native enzyme. Tetragonal crystals suitable for X-ray crystallography studies were obtained; they were improved by recrystallization and found to diffract to 2.0 A resolution using synchrotron radiation. The crystals belong to the tetragonal space group I422 with unit cell dimensions a = b = 154.7 A, c = 124.9 A, giving a Vm of 2.73 A3 per Da (estimated 60% solvent) for a single molecule of recombinant BChE in the asymmetric unit. The crystal structure of butyrylcholinesterase will help elucidate unsolved issues concerning cholinesterase mechanisms in general. PMID- 11856323 TI - Dematin interacts with the Ras-guanine nucleotide exchange factor Ras-GRF2 and modulates mitogen-activated protein kinase pathways. AB - Erythroid dematin is a major component of red blood cell junctional complexes that link the spectrin-actin cytoskeleton to the overlying plasma membrane. Transcripts of dematin are widely distributed including human brain, heart, lung, skeletal muscle, and kidney. In vitro, dematin binds and bundles actin filaments in a phosphorylation-dependent manner. The primary structure of dematin consists of a C-terminal domain homologous to the 'headpiece' domain of villin, an actin binding protein of the brush border cytoskeleton. Except filamentous actin, no other binding partners of dematin have been identified. To investigate the physiological function of dematin, we employed the yeast two-hybrid assay to identify dematin-interacting proteins in the adult human brain. Here, we show that dematin interacts with the guanine nucleotide exchange factor Ras-GRF2 by yeast two-hybrid assay, and this interaction is further confirmed by blot overlay, surface plasmon resonance, co-transfection, and co-immunoprecipitation assays. Human Ras-GRF2 is expressed in a variety of tissues and, similar to other guanine nucleotide exchange factors (GEFs), displays anchorage independent growth in soft agar. Co-transfection and immunoblotting experiments revealed that dematin blocks transcriptional activation of Jun by Ras-GRF2 and activates ERK1 via a Ras-GRF2 independent pathway. Because much of the present evidence has centered on the identification of the Rho family of GTPases as key regulators of the actin cytoskeleton, the direct association between dematin and Ras-GRF2 may provide an alternate mechanism for regulating the activation of Rac and Ras GTPases via the actin cytoskeleton. PMID- 11856324 TI - Substrate recognition and fidelity of strand joining by an archaeal DNA ligase. AB - We have previously identified a DNA ligase (LigTk) from a hyperthermophilic archaeon, Thermococcus kodakaraensis KOD1. The enzyme is the only characterized ATP-dependent DNA ligase from a hyperthermophile, and allows the analysis of enzymatic DNA ligation reactions at temperatures above the melting point of the substrates. Here we have focused on the interactions of LigTk with various DNA substrates, and its specificities toward metal cations. LigTk could utilize Mg2+, Mn2+, Sr2+ and Ca2+ as a metal cation, but not Co2+, Zn2+, Ni2+, or Cu2+. The enzyme displayed typical Michaelis-Menten steady-state kinetics with an apparent Km of 1.4 microm for nicked DNA. The kcat value of the enzyme was 0.11*s-1. Using various 3' hydroxyl group donors (L-DNA) and 5' phosphate group donors (R-DNA), we could detect ligation products as short as 16 nucleotides, the products of 7 + 9 nucleotide or 8 + 8 nucleotide combinations at 40 degrees C. An elevation in temperature led to a decrease in reaction efficiency when short oligonucleotides were used, suggesting that the formation of a nicked, double-stranded DNA substrate preceded enzyme-substrate recognition. LigTk was not inhibited by the addition of excess duplex DNA, implying that the enzyme did not bind strongly to the double-stranded ligation product after nick-sealing. In terms of reaction fidelity, LigTk was found to ligate various substrates with mismatched base pairing at the 5' end of the nick, but did not show activity towards the 3' mismatched substrates. LigTk could not seal substrates with a 1-nucleotide or 2 nucleotide gap. Small amounts of ligation products were detected with DNA substrates containing a single nucleotide insertion, relatively more with the 5' insertions. The results revealed the importance of proper base-pairing at the 3' hydroxyl side of the nick for the ligation reaction by LigTk. PMID- 11856325 TI - Spermosin, a trypsin-like protease from ascidian sperm: cDNA cloning, protein structures and functional analysis. AB - We have previously reported that two trypsin-like enzymes, acrosin and spermosin, play key roles in sperm penetration through the vitelline coat of the ascidian (Urochordata) Halocynthia roretzi [Sawada et al. (1984), J. Biol. Chem. 259, 2900 2904; Sawada et al. (1984), Dev. Biol. 105, 246-249]. Here, we show the amino acid sequence of the ascidian preprospermosin, which is deduced from the nucleotide sequence of the isolated cDNA clone. The isolated ascidian preprospermosin cDNA consisted of 1740 nucleotides, and an open reading frame encoding 388 amino acids, which corresponds to a molecular mass of 41 896 Da. By sequence alignment, it was suggested that His178, Asp230 and Ser324 make up a catalytic triad and that ascidian spermosin be classified as a novel trypsin family member. The mRNA of preprospermosin is specifically expressed in ascidian gonads but not in other tissues. Purified spermosin consists of 33- and 40-kDa bands as determined by SDS/PAGE under nonreducing conditions. The 40-kDa spermosin consists of a heavy chain (residues 130-388) and a long light chain designated L1 (residues 23-129), whereas the 33-kDa spermosin includes the same heavy chain and a shorter light chain designated L2 (residues 97-129). The L1 chain contains a proline-rich region, designated L1(DeltaL2) which is lacking in L2. Investigation with the glutathione-S-transferase (GST)-spermosin-light-chain fusion proteins, including GST-L1, GST-L2, and GST-L1(DeltaL2), revealed that the proline-rich region in the L1 chain binds to the vitelline coat of ascidian eggs. Thus, we propose that sperm spermosin is a novel trypsin-like protease that binds to the vitelline coat and also plays a part in penetration of sperm through the vitelline coat during ascidian fertilization. PMID- 11856326 TI - Early signaling events induced by 280-nm UV irradiation. AB - The depletion of stratospheric ozone results in increased UV (ultraviolet) light below 300 nm, and has significant effects on biological systems. To better understand the effects of UV in this range, early signaling events induced by monochromatic UV light were investigated using the chicken B cell line DT40 and mutants lacking protein tyrosine kinases (PTKs). Among MAP kinase family proteins, P38 MAP kinase (P38) was selectively and immediately activated by 280 nm UV light in cultured DT40 cells. Activation of P38 was completely inhibited in cells deficient in Lyn and Btk. Introduction of wild-type Btk, but not kinase inactive Btk, restored the P38 activation. In contrast, P38 activation was not affected in Syk-deficient cells. Tyrosine phosphorylation of Lyn was induced by 280 nm UV light, and pretreatment of cells with orthovanadate, an inhibitor of protein tyrosine phosphatase (PTP), enhanced both Lyn phosphorylation and P38 activation. These results show that Lyn and Btk are upstream regulators of the P38 signaling pathway activated by 280 nm UV light and that the triggering event likely involves inactivation of PTP. Furthermore, cell death induced by 280 nm UV irradiation were augmented by Btk depletion or a specific inhibitor for P38, and partially blocked in Lyn-deficient cells, suggesting that the Lyn-Btk-P38 pathway promotes cell survival while other Lyn pathways stimulate cell death. PMID- 11856327 TI - Maturation of Pichia pastoris-derived recombinant pro-Der p 1 induced by deglycosylation and by the natural cysteine protease Der p 1 from house dust mite. AB - The mature cysteine protease from Dermatophgoides pteronyssinus, Der p 1, is a major house dust mite allergen. Its enzymatic activity has been shown to have pro inflammatory effects that could also negatively influence efficacy of allergen specific immunotherapy. The aim of this study was to express recombinant pro-Der p 1 (rpro-Der p 1) in the yeast Pichia pastoris and to study its maturation. Expression was achieved at a concentration ranging from 45 mg.L-1 (methanol induced expression) to 168 mg.L-1 (constitutive expression). No significant spontaneous maturation of the secreted proenzyme was observed. rpro-Der p 1 with a sequence-based molecular mass of 34 kDa was hyperglycosylated by the yeast, migrating at 50-60 kDa on SDS/PAGE. Compared with its natural counterpart (nDer p 1), the recombinant proenzyme demonstrated decreased IgE reactivity, resulting in a 30-fold lower capacity to induce histamine release from human basophils. Decreased immunoreactivity was also shown by competitive RIA and sandwich ELISA with Der p 1-specific antibody reagents. CD spectra of rpro-Der p 1 and nDer p 1 revealed significant structural differences. Deglycosylation of rpro-Der p 1 with endoglycosidase H resulted in a decrease in apparent molecular mass from 50 kDa to 34 kDa, but did not affect nDer p 1. On removal of N-glycans from rpro-Der p 1, which harbours two putative N-glycosylation sites in both propeptide and mature sequence, the mature rDer p 1 appeared. This suggests that hyperglycosylation hampers spontaneous maturation. Maturation of the recombinant pro-enzyme was also achieved by addition of the active natural cysteine protease, nDer p 1. In conclusion, high-level expression of rpro-Der p 1 in P. pastoris results in a stable hypoallergenic proenzyme with potential for use in allergen specific immunotherapy. PMID- 11856328 TI - Characterization and synthetic applications of recombinant AtNIT1 from Arabidopsis thaliana. AB - The nitrilase AtNIT1 from Arabidopsis thaliana was overexpressed in Escherichia coli with an N-terminal His6 tag and purified by zinc chelate affinity chromatography in a single step almost to homogeneity in a 68% yield with a specific activity of 34.1 U.mg-1. The native enzyme (approximately 450 kDa) consists of 11-13 subunits (38 kDa). The temperature optimum was determined to be 35 degrees C and a pH optimum of 9 was found. Thus, recombinant AtNIT1 resembles in its properties the native enzyme and the nitrilase from Brassica napus. The stability of AtNIT1 could be significantly improved by the addition of dithiothreitol and EDTA. The substrate range of AtNIT1 differs considerably from those of bacterial nitrilases. Aliphatic nitriles are the most effective substrates, showing increasing rates of hydrolysis with increasing size of the residues, as demonstrated in the series butyronitrile, octanenitrile, phenylpropionitrile. In comparison with 3-indolylacetonitrile, the rate of hydrolysis of 3-phenylpropionitrile is increased by a factor of 330, and the Km value is reduced by a factor of 23. With the exception of fluoro, substituents in the alpha position to the nitrile function completely inhibit the hydrolysis. PMID- 11856329 TI - Fish otolith contains a unique structural protein, otolin-1. AB - A collagen-like protein was identified from the otoliths of the chum salmon, Oncorhynchus keta. The otolith, composed mainly of calcium carbonate with small amount of organic matrices, is formed in the inner ear and serves as a part of the hearing and balance systems. Although the organic matrices may play important roles in the growth of otolith, little is known about their chemical nature and physiological function. In this study, a major organic component of the otolith, designated otolin-1, which may serve as a template for calcification, was purified. The sequences of two tryptic peptides from otolin-1 revealed high homology with parts of a saccular collagen which had been described previously [Davis, J.G., Oberholtzer, J.C., Burns, F.R. & Greene, M.I. (1995) Science 267, 1031-1034]. Cloning of a cDNA coding for otolin-1 revealed that the deduced amino acid sequence contained a collagenous domain in the central part of the protein. Although collagen is the most abundant structural protein in the animal body, otolin-1 mRNA was expressed specifically in the sacculus. Immunohistochemical studies showed that otolin-1 is synthesized in the transitional epithelium and transferred to the otolith and otolithic membrane. This is the first report concerning characterization of a structural protein containing many tandem repeats of the sequence, Gly-Xaa-Yaa, typical for collagen from the biomineral composed of calcium carbonate. PMID- 11856330 TI - Anti-(Raf-1) RNA aptamers that inhibit Ras-induced Raf-1 activation. AB - RNA aptamers with affinity for the Ras-binding domain (RBD) of Raf-1 were isolated from a degenerate pool by in vitro selection. These aptamers efficiently inhibited the Ras interaction with the Raf-1 RBD, and also inhibited Ras-induced Raf-1 activation in a cell-free system. The RNA aptamer with the most potent inhibitory effect specifically inhibited the Ras-Raf-1 interaction and had no affinity for the RBD of the RGL protein, a homolog of the Ral GDP dissociation stimulator. Although the aptamer was capable of binding to the B-Raf RBD, the RNA did not inhibit the interaction between Ras and the B-Raf RBD. Enzymatic and chemical probing experiments indicated that the aptamer was folded into a pseudoknot structure, and some loop regions of the pseudoknot were located at the binding interface for the Raf-1 RBD. PMID- 11856331 TI - Crystal structure of a staphylokinase: variant a model for reduced antigenicity. AB - Staphylokinase (SAK) is a 15.5-kDa protein from Staphylococcus aureus that activates plasminogen by forming a 1 : 1 complex with plasmin. Recombinant SAK has been shown in clinical trials to induce fibrin-specific clot lysis in patients with acute myocardial infarction. However, SAK elicits high titers of neutralizing antibodies. Biochemical and protein engineering studies have demonstrated the feasibility of generating SAK variants with reduced antigenicity yet intact thrombolytic potency. Here, we present X-ray crystallographic evidence that the SAK(S41G) mutant may assume a dimeric structure. This dimer model, at 2.3-A resolution, could explain a major antigenic epitope (residues A72-F76 and residues K135-K136) located in the vicinity of the dimer interface as identified by phage-display. These results suggest that SAK antigenicity may be reduced by eliminating dimer formation. We propose several potential mutation sites at the dimer interface that may further reduce the antigenicity of SAK. PMID- 11856332 TI - Human milk provides peptides highly stimulating the growth of bifidobacteria. AB - The large intestine of breast-fed infants is colonized predominantly by bifidobacteria, which have a protective effect against acute diarrhea. In this study we report for the first time the identification of human milk peptides that selectively stimulate the growth of bifidobacteria. Several bifidogenic peptides were purified chromatographically from pepsin-treated human milk and identified as proteolytically generated fragments from the secretory component of the soluble polyimmunoglobulin receptor and lactoferrin; both of these proteins exhibit antimicrobial effects. Hydrolysis of the identified peptides with the gastrointestinal proteases pepsin, trypsin and chymotrypsin did not lead to the loss of bifidogenic activity, indicating their potential function in vivo. Sequential comparison revealed a similar structural motif within the identified peptides. A correspondingly designed small peptide (prebiotic lactoferrin-derived peptide-I, PRELP-I) was found to stimulate the growth of bifidobacteria as effectively as the native peptides. The combination of antimicrobial and bifidobacterial growth stimulatory activity in human milk proteins leads to highly specific compounds capable of regulating the microbial composition of infants' large intestine. PMID- 11856333 TI - Functional site of endogenous phospholipase A2 inhibitor from python serum. AB - The functional site of 'phospholipase A2 inhibitor from python' (PIP) was predicted based on the hypothesis of proline brackets. Using different sources of secretory phospholipase A2 (sPLA2s) as enzyme, and [3H]arachidonate-labelled Escherichia coli as substrate, short synthetic peptides representing the proposed site were examined for their secretory phospholipase A2 (sPLA2) inhibitory activity. A decapeptide P-PB.III proved to be the most potent of the tested peptides in inhibiting sPLA2 enzymatic activity in vitro, and exhibited striking anti-inflammatory effects in vivo in a mouse paw oedema model. P-PB.III inhibited the enzymatic activity of class I, II and III PLA2s, including that of human synovial fluid from arthritis patients. When tested by ELISA, biotinylated P PB.III interacted positively with various PLA2s, suggesting that the specific region of PIP corresponding to P-PB.III, is likely to be involved in the PLA2-PLI interaction. The effect of P-PB.III on the peritoneal inflammatory response after surgical trauma in rats was also examined. P-PB.III effectively reduced the extent of postsurgical peritoneal adhesions as compared to controls. sPLA2 levels at seventh postoperative day in the peritoneal tissue of P-PB.III-treated rats were also significantly reduced (P < 0.05) in comparison to those of the untreated controls. The present results shed additional insight on the essential structural elements for PLA2 binding, and may be useful as a basis for the design of novel therapeutic agents. PMID- 11856334 TI - Substrate recognition by three family 13 yeast alpha-glucosidases. AB - Important hydrogen bonding interactions between substrate OH-groups in yeast alpha-glucosidases and oligo-1,6-glucosidase from glycoside hydrolase family 13 have been identified by measuring the rates of hydrolysis of methyl alpha isomaltoside and its seven monodeoxygenated analogs. The transition-state stabilization energy, DeltaDeltaG, contributed by the individual OH-groups was calculated from the activities for the parent and the deoxy analogs, respectively, according to DeltaDeltaG = -RT ln[(Vmax/Km)analog/(Vmax/Km)parent]. This analysis of the energetics gave DeltaDeltaG values for all three enzymes ranging from 16.1 to 24.0 kJ.mol-1 for OH-2', -3', -4', and -6', i.e. the OH groups of the nonreducing sugar ring. These OH-groups interact with enzyme via charged hydrogen bonds. In contrast, OH-2 and -3 of the reducing sugar contribute to transition-state stabilization, by 5.8 and 4.1 kJ.mol-1, respectively, suggesting that these groups participate in neutral hydrogen bonds. The OH-4 group is found to be unimportant in this respect and very little or no contribution is indicated for all OH-groups of the reducing-end ring of the two alpha-glucosidases, probably reflecting their exposure to bulk solvent. The stereochemical course of hydrolysis by these three members of the retaining family 13 was confirmed by directly monitoring isomaltose hydrolysis using 1H NMR spectroscopy. Kinetic analysis of the hydrolysis of methyl 6-S-ethyl-alpha isomaltoside and its 6-R-diastereoisomer indicates that alpha-glucosidase has 200 fold higher specificity for the S-isomer. Substrate molecular recognition by these alpha-glucosidases are compared to earlier findings for the inverting, exo acting glucoamylase from Aspergillus niger and a retaining alpha-glucosidase of glycoside hydrolase family 31, respectively. PMID- 11856336 TI - Coordinated action of protein tyrosine phosphatases in insulin signal transduction. AB - Insulin is the principal regulatory hormone involved in the tight regulation of fuel metabolism. In response to blood glucose levels, it is secreted by the beta cells of the pancreas and exerts its effects by binding to cell surface receptors that are present on virtually all cell types and tissues. In humans, perturbations in insulin function and/or secretion lead to diabetes mellitus, a severe disorder primarily characterized by an inability to maintain blood glucose homeostasis. Furthermore, it is estimated that 90-95% of diabetic patients exhibit resistance to insulin action. Thus an understanding of insulin signal transduction and insulin resistance at the molecular level is crucial to the understanding of the pathogenesis of this disease. The insulin receptor (IR) is a transmembrane tyrosine kinase that becomes activated upon ligand binding. Consequently, the receptor and its downstream substrates become tyrosine phosphorylated. This activates a series of intracellular signaling cascades which coordinately initiate the appropriate biological response. One important mechanism by which insulin signaling is regulated involves the protein tyrosine phosphatases (PTPs), which may either act on the IR itself and/or its substrates. Two well characterized examples include leuckocyte antigen related (LAR) and protein tyrosine phosphatase-1B (PTP-1B). The present review will discuss the current knowledge of these two and other potential PTPs involved in the insulin signaling pathway. PMID- 11856337 TI - Regulation of stress-activated protein kinase signaling pathways by protein phosphatases. AB - Stress-activated protein kinase (SAPK) signaling plays essential roles in eliciting adequate cellular responses to stresses and proinflammatory cytokines. SAPK pathways are composed of three successive protein kinase reactions. The phosphorylation of SAPK signaling components on Ser/Thr or Thr/Tyr residues suggests the involvement of various protein phosphatases in the negative regulation of these systems. Accumulating evidence indicates that three families of protein phosphatases, namely the Ser/Thr phosphatases, the Tyr phosphatases and the dual specificity Ser/Thr/Tyr phosphatases regulate these pathways, each mediating a distinct function. Differences in substrate specificities and regulatory mechanisms for these phosphatases form the molecular basis for the complex regulation of SAPK signaling. Here we describe the properties of the protein phosphatases responsible for the regulation of SAPK signaling pathways. PMID- 11856338 TI - Novel protein phosphatases in yeast. AB - During the last decade several novel yeast genes encoding proteins related to the PPP family of Ser/Thr protein phosphatases have been discovered and their functional characterization initiated. Most of these novel phosphatases display intriguing structural features and/or are involved in a number of important functions, such as cell cycle regulation, protein synthesis and maintenance of cellular integrity. While in some cases these genes appear to be restricted to fungi, in others similar proteins can be found in higher eukaryotes. This review will summarize the latest advances in our understanding about how these phosphatases are regulated and fulfil their functions in the yeast cell. PMID- 11856339 TI - The function of methyl-menaquinone-6 and polysulfide reductase membrane anchor (PsrC) in polysulfide respiration of Wolinella succinogenes. AB - Wolinella succinogenes grows by oxidative phosphorylation with polysulfide as terminal electron acceptor and either H2 or formate as electron donor (polysulfide respiration). The function of the respiratory chains catalyzing these reactions was investigated. Proteoliposomes containing polysulfide reductase (Psr) and either hydrogenase or formate dehydrogenase isolated from the membrane fraction of Wolinella succinogenes catalyzed polysulfide respiration, provided that methyl-menaquinone-6 isolated from W. succinogenes was also present. The specific activities of electron transport were commensurate with those of the bacterial membrane fraction. Using site-directed mutagenesis, certain residues were substituted in PsrC, the membrane anchor of polysulfide reductase. Replacement of Y23, D76, Y159, D218, E225 or R305 caused nearly full inhibition of polysulfide respiration without affecting the activity of Psr, which was still bound to the membrane. These residues are predicted to be located in hydrophobic helices of PsrC, or next to them. Substitution of 13 other residues of PsrC either caused partial inhibition ofblankpolysulfide respiration or had no effect. The function of methyl-menaquinone-6, which is thought to be bound to PsrC, is discussed. PMID- 11856340 TI - Exploring the primary electron acceptor (QA)-site of the bacterial reaction center from Rhodobacter sphaeroides. Binding mode of vitamin K derivatives. AB - The functional replacement of the primary ubiquinone (QA) in the photosynthetic reaction center (RC) from Rhodobacter sphaeroides with synthetic vitamin K derivatives has provided a powerful tool to investigate the electron transfer mechanism. To investigate the binding mode of these quinones to the QA binding site we have determined the binding free energy and charge recombination rate from QA(-) to D+ (kAD) of 29 different 1,4-naphthoquinone derivatives with systematically altered structures. The most striking result was that none of the eight tested compounds carrying methyl groups in both positions 5 and 8 of the aromatic ring exhibited functional binding. To understand the binding properties of these quinones on a molecular level, the structures of the reaction center naphthoquinone complexes were predicted with ligand docking calculations. All protein--ligand structures show hydrogen bonds between the carbonyl oxygens of the quinone and AlaM260 and HisM219 as found for the native ubiquinone-10 in the X-ray structure. The center-to-center distance between the naphthoquinones at QA and the native ubiquinone-10 at QB (the secondary electron acceptor) is essentially the same, compared to the native structure. A detailed analysis of the docking calculations reveals that 5,8-disubstitution prohibits binding due to steric clashes of the 5-methyl group with the backbone atoms of AlaM260 and AlaM249. The experimentally determined binding free energies were reproduced with an rmsd of approximately 4 kJ x mol(-1) in most cases providing a valuable tool for the design of new artificial electron acceptors and inhibitors. PMID- 11856341 TI - Myristyl and palmityl acylation of pI 5.1 carboxylesterase from porcine intestine and liver. AB - Immunoblotting analyses revealed the presence of carboxylesterase in the porcine small intestine, liver, submaxillary and parotid glands, kidney cortex, lungs and cerebral cortex. In the intestinal mucosa, the pI 5.1 enzyme was detected in several subcellular fractions including the microvillar fraction. Both fatty monoacylated and diacylated monomeric (F1), trimeric (F3) and tetrameric (F4) forms of the intestinal protein were purified here for the first time by performing hydrophobic chromatography and gel filtration. The molecular mass of these three enzymatic forms was estimated to be 60, 180 and 240 kDa, respectively, based on size-exclusion chromatography and SDS/PAGE analysis. The existence of a covalent attachment linking palmitate and myristate to porcine intestinal carboxylesterase (PICE), which was suggested by the results of gas liquid chromatography (GLC) experiments in which the fatty acids resulting from alkali treatment of the protein forms were isolated, was confirmed here by the fact that [3H]palmitic and [3H]myristic acids were incorporated into porcine enterocytes and hepatocytes in cell primary cultures. Besides these two main fatty acids, the presence of oleic, stearic, and arachidonic acids was also detected by GLC and further confirmed by performing radioactivity counts on the 3H-labelled PICE forms after an immunoprecipitation procedure using specific polyclonal antibodies, followed by a SDS/PAGE separation step. Unlike the F1 and F4 forms, which were both myristoylated and palmitoylated, the F3 form was only palmitoylated. The monomeric, trimeric and tetrameric forms of PICE were all able to hydrolyse short chain fatty acids containing glycerides, as well as phorbol esters. The broad specificity of fatty acylated carboxylesterase is discussed in terms of its possible involvement in the metabolism of ester-containing xenobiotics and signal transduction. PMID- 11856342 TI - The SK-N-MC cell line expresses an orexin binding site different from recombinant orexin 1-type receptor. AB - Orexin A and B (also known as hypocretins), two recently discovered neuropeptides, play an important role in food intake, sleep/wake cycle and neuroendocrine functions. Orexins are endogenous ligands of two G-protein-coupled receptors, termed OX1 and OX2. This work presents the first short orexin A and B analogues, orexin A 23-33 and orexin B 18-28, with high affinity (119 +/- 49 and 49 +/- 23 nm) for OX1 receptors expressed on SK-N-MC cells and indicates the importance of the C-terminal part of the orexin peptides for this ligand-receptor interaction. However, these C-terminal fragments of orexin did not displace the 125I-labelled orexin B from the recombinant orexin 1 receptor stably expressed in Chinese hamster ovary cells. To examine the role of the shortened orexin A 23-33 in feeding, its effects in mimicking or antagonizing the effects of orexin A were studied in rats after administration via the lateral hypothalamus. In contrast with orexin A, which potently induced feeding up to 4 h after administration, orexin A 23-33 neither induced feeding nor inhibited orexin A-induced feeding. Modafinil (Vigil), which was shown earlier to activate orexin neurons, displayed binding neither to the orexin receptor expressed on SK-N-MC cells nor to the recombinant orexin 1 receptor, which indicates that modafinil displays its antinarcoleptic action via another yet unknown mechanism. PCR and subsequent sequencing revealed expression of the full-length orexin 1 receptor mRNA in SK-N MC and NT-2 cells. Interestingly, sequencing of several cDNA clones derived from RNA of both SK-N-MC and NT-2 cells differed from the published nucleotide sequence at position 1375. Amino acid prediction of this A -->G change results in an isoleucine --> valine substitution at the protein level, which may provide evidence for an editing process. PMID- 11856343 TI - A novel gain-of-function mutation of the integrin alpha2 VWFA domain. AB - Integrin alpha2beta1 is the major receptor for collagens in human tissues, being involved in cell adhesion and the control of collagen and collagenase gene expression. The collagen binding site of alpha2beta1 has been localized to the alpha2 von Willebrand Factor type A (VWFA) domain (A-domain or I-domain) and the residues responsible for the interaction with collagen have been mapped. We report a study of alpha2 VWFA domain in which residue E318, which lies outside the collagen binding site, is mutated to tryptophan, showing that this is a gain of-function mutation. Recombinant alpha2-E318W VWFA domain showed elevated and specific binding to collagen I compared with the wild-type. Side chain hydrophobicity was important for the gain-of-function as elevated binding was seen with E318I and E318Y, but not with E318R. The E318W mutation had additional effects on VWFA domain properties as alpha2-E318W VWFA domain differed from the wild-type in its cation preferences for ligand binding and in binding to monoclonal antibody JA203, which bound at a site distal to E318. The gain-of function effect was not restricted to binding to collagen I as alpha2-E318W also showed elevated binding to collagen IV, collagen I C-propeptide, laminin and E cadherin. Binding to these ligands was inhibited by collagen peptide containing the GFOGER motif, indicating that these bound to the VWFA domain by a similar mechanism to collagen I. These data indicate that residue E318 plays a novel and important role in modulating alpha2 VWFA domain--ligand binding and may be involved in the conformational changes associated with its regulation. PMID- 11856345 TI - The solution structure of gomesin, an antimicrobial cysteine-rich peptide from the spider. AB - Gomesin is the first peptide isolated from spider exhibiting antimicrobial activities. This highly cationic peptide is composed of 18 amino-acid residues including four cysteines forming two disulfide linkages. The solution structure of gomesin has been determined using proton two-dimensional NMR (2D-NMR) and restrained molecular dynamics calculations. The global fold of gomesin consists in a well-resolved two-stranded antiparallel betasheet connected by a noncanonical betaturn. A comparison between the structures of gomesin and protegrin-1 from porcine and androctonin from scorpion outlines several common features in the distribution of hydrophobic and hydrophilic residues. The N- and C-termini, the betaturn and one face of the betasheet are hydrophilic, but the hydrophobicity of the other face depends on the peptide. The similarities suggest that the molecules interact with membranes in an analogous manner. The importance of the intramolecular disulfide bridges in the biological activity of gomesin is being investigated. PMID- 11856344 TI - SMAP-29 has two LPS-binding sites and a central hinge. AB - The CD spectra of SMAP-29, an antimicrobial peptide from sheep, showed disordered structure in aqueous buffers, and significant helicity in membrane-like environments, including SDS micelles, lipopolysaccharide (LPS) dispersions, and trifluoroethanol buffer systems. A structure determined by NMR in 40% perdeuterated trifluoroethanol indicated that residues 8-17 were helical, residues 18-19 formed a hinge, and residues 20-28 formed an ordered, hydrophobic segment. SMAP-29 was flexible in 40% trifluoroethanol, forming two sets of conformers that differed in the relative orientation of the N-terminal domain. We used a chromogenic Limulus assay to determine the EC50 of the peptide (the concentration that bound 50% of the added LPS). Studies with full-length and truncated SMAP-29 molecules revealed that each end of the holopeptide contained an LPS-binding domain. The higher affinity LPS-binding domain was situated in the flexible N-terminal portion. LPS binding to full-length SMAP-29 showed positive cooperativity, so the EC50 of the peptide (2.6 microm) was considerably lower than that of the individual LPS-binding domains. LPS-binding studies with a mixture of truncated peptides revealed that this cooperativity was primarily intramolecular (i.e. involving the N- and C-terminal LPS-binding sites of the same peptide molecule). CAP-18[106 -142], an antimicrobial cathelicidin peptide of rabbits, resembled SMAP-29 in that it contained N- and C-terminal LPS-binding domains, had an EC50 of 2.5 microm, and bound LPS with positive cooperativity. We conclude that the presence of multiple binding sites that function cooperatively allow peptides such as SMAP-29 and CAP-18 to bind LPS with high affinity. PMID- 11856347 TI - Phosphorylation and dephosphorylation of histidine residues in proteins. AB - Protein phosphorylation is a key mechanism for intracellular signal transduction in both prokaryotic and eukaryotic cells. Vertebrate proteins are prevalently phosphorylated on side chains that contain a hydroxyl group, such as serine, threonine and tyrosine residues. In the past decade, however, an increasing number of examples of histidine phosphorylation has been described. Because acid treatment of phosphoproteins during purification and detection of phosphoamino acid analysis is routine, O-phosphomonoesters have been studied more often, and the existence of acid-labile phosphates has been largely overlooked. The latter class of N-phosphoamidates may well be more widespread than is generally believed, even though the O-phosphates remain the major class in terms of quantity and extent of distribution in proteins. Phosphohistidine currently is estimated to be 10- to 100-fold more abundant than phosphotyrosine, but less abundant than phosphoserine [Matthews, H.R. (1995) Pharmac. Ther. 67, 323-350.]. This minireview briefly summarizes the extensive knowledge of the key mechanisms and functions of phosphohistidine in bacteria. It also describes the still limited, yet increasing, data from homologs of the bacterial two-component system. Finally, novel mechanisms of phosphorylation and dephosphorylation of histidine residues not related to the two-component system are described. PMID- 11856346 TI - GPI-microdomains (membrane rafts) and signaling of the multi-chain interleukin-2 receptor in human lymphoma/leukemia T cell lines. AB - Subunits (alpha, beta and gamma) of the interleukin-2 receptor complex (IL-2R) are involved in both proliferative and activation-induced cell death (AICD) signaling of T cells. In addition, the signaling beta and gamma chains are shared by other cytokines (e.g. IL-7, IL-9, IL-15). However, the molecular mechanisms responsible for recruiting/sorting the alpha chains to the signaling chains at the cell surface are not clear. Here we show, in four cell lines of human adult T cell lymphoma/leukemia origin, that the three IL-2R subunits are compartmented together with HLA glycoproteins and CD48 molecules in the plasma membrane, by means of fluorescence resonance energy transfer (FRET), confocal microscopy and immuno-biochemical techniques. In addition to the beta and gamma(c) chains constitutively expressed in detergent-resistant membrane fractions (DRMs) of T cells, IL-2Ralpha (CD25) was also found in DRMs, independently of its ligand occupation. Association of CD25 with rafts was also confirmed by its colocalization with GM-1 ganglioside. Depletion of membrane cholesterol using methyl-beta-cyclodextrin substantially reduced co-clustering of CD25 with CD48 and HLA-DR, as well as the IL-2 stimulated tyrosine-phosphorylation of STATs (signal transducer and activator of transcription). These data indicate a GPI microdomain (raft)-assisted recruitment of CD25 to the vicinity of the signaling beta and gamma(c) chains. Rafts may promote rapid formation of a high affinity IL 2R complex, even at low levels of IL-2 stimulus, and may also form a platform for the regulation of IL-2 induced signals by GPI-proteins (e.g. CD48). Based on these data, the integrity of these GPI-microdomains seems critical in signal transduction through the IL-2R complex. PMID- 11856348 TI - Evidence that a eukaryotic-type serine/threonine protein kinase from Mycobacterium tuberculosis regulates morphological changes associated with cell division. AB - A eukaryotic-type protein serine/threonine kinase, PknA, was cloned from Mycobacterium tuberculosis strain H37Ra. Sequencing of the clone indicated 100% identity with the published pknA sequence of M. tuberculosis strain H37Rv. PknA fused to maltose-binding protein was expressed in Escherichia coli; it exhibited a molecular mass of approximately 97 kDa. The fusion protein was purified from the soluble fraction by affinity chromatography using amylose resin. In vitro kinase assays showed that the autophosphorylating ability of PknA is strictly magnesium/manganese-dependent, and sodium orthovanadate can inhibit this activity. Phosphoamino-acid analysis indicated that PknA phosphorylates at serine and threonine residues. PknA was also able to phosphorylate exogenous substrates, such as myelin basic protein and histone. A comparison of the nucleotide-derived amino-acid sequence of PknA with that of functionally characterized prokaryotic serine/threonine kinases indicated its possible involvement in cell division/differentiation. Protein--protein interaction studies revealed that PknA is capable of phosphorylating at least a approximately 56-kDa soluble protein from E. coli. Scanning electron microscopy showed that constitutive expression of this kinase resulted in elongation of E. coli cells, supporting its regulatory role in cell division. PMID- 11856349 TI - The effect of amino-acid substitutions I112P, D147E and K152N in CYP11B2 on the catalytic activities of the enzyme. AB - By replacing specific amino acids at positions 112, 147 and 152 of the human aldosterone synthase (CYP11B2) with the corresponding residues from human, mouse or rat 11beta-hydroxylase (CYP11B1), we have been able to investigate whether these residues belong to structural determinants of individual enzymatic activities. When incubated with 11-deoxycorticosterone (DOC), the 11beta hydroxylation activity of the mutants was most effectively increased by combining D147E and I112P (sixfold increase). The two substitutions displayed an additive effect. The same tendency can be observed when using 11-deoxycortisol as a substrate, although the effect is less pronounced. The second step of the CYP11B2 dependent DOC conversion, the 18-hydroxylation activity, was not as strongly increased as the 11beta-hydroxylation potential. Activity was unaffected by D147E, whereas the single mutant I112P displayed the most pronounced activation (70% enhancement), thus causing different increasing effects on the first two enzymatic reaction steps. A slightly enhanced aldosterone synthesis from DOC could be measured due to increased levels of the intermediates. However, the 18 oxidation activity of all the mutants, except for I112S and D147E, was slightly reduced. The strongly enhanced 18-hydroxycorticosterone and aldosterone formation observed in the mutants provides important information on a possible role of such amino-acid replacements in the development of essential hypertension. Furthermore, the results indicate the possibility of a differential as well as independent modification of CYP11B2 reaction steps. The combination of functional data and computer modelling of CYP11B2 suggests an indirect involvement of residue 147 in the regulation of CYP11B isoform specific substrate conversion due to its location on the protein surface. In addition, the results indicate the functional significance of amino-acid 112 in the putative substrate access channel of human CYP11B2. Thus, we present the first example of substrate recognition and conversion being attributed to the N-terminal part of human CYP11B2. PMID- 11856350 TI - Stabilization of a (betaalpha)8-barrel protein by an engineered disulfide bridge. AB - The aim of this study was to increase the stability of the thermolabile (betaalpha)8-barrel enzyme indoleglycerol phosphate synthase from Escherichia coli by the introduction of disulfide bridges. For the design of such variants, we selected two out of 12 candidates, in which newly introduced cysteines potentially form optimal disulfide bonds. These variants avoid short-range connections, substitutions near catalytic residues, and crosslinks between the new and the three parental cysteines. The variant linking residues 3 and 189 fastens the N-terminus to the (betaalpha)8-barrel. The rate of thermal inactivation at 50 degrees C of this variant with a closed disulfide bridge is 65 fold slower than that of the reference dithiol form, but only 13-fold slower than that of the parental protein. The near-ultraviolet CD spectrum, the reactivity of parental buried cysteines with Ellman's reagent as well as the decreased turnover number indicate that the protein structure is rigidified. To confirm these data, we have solved the X-ray structure to 2.1-A resolution. The second variant was designed to crosslink the terminal modules betaalpha1 and betaalpha8. However, not even the dithiol form acquired the native fold, possibly because one of the targeted residues is solvent-inaccessible in the parental protein. PMID- 11856351 TI - Concentration-dependent reversible activation-inhibition of human butyrylcholinesterase by tetraethylammonium ion. AB - Tetraalkylammonium (TAA) salts are well known reversible inhibitors of cholinesterases. However, at concentrations around 10 mm, they have been found to activate the hydrolysis of positively charged substrates, catalyzed by wild-type human butyrylcholinesterase (EC 3.1.1.8) [Erdoes, E.G., Foldes, F.F., Zsigmond, E.K., Baart, N. & Zwartz, J.A. (1958) Science 128, 92]. The present study was undertaken to determine whether the peripheral anionic site (PAS) of human BuChE (Y332, D70) and/or the catalytic substrate binding site (CS) (W82, A328) are involved in this phenomenon. For this purpose, the kinetics of butyrylthiocholine (BTC) hydrolysis by wild-type human BuChE, by selected mutants and by horse BuChE was carried out at 25 degreeC and pH 7.0 in the presence of tetraethylammonium (TEA). It appears that human enzymes with more intact structure of the PAS show more prominent activation phenomenon. The following explanation has been put forward: TEA competes with the substrate at the peripheral site thus inhibiting the substrate hydrolysis at the CS. As the inhibition by TEA is less effective than the substrate inhibition itself, it mimics activation. At the concentrations around 40 mm, well within the range of TEA competition at both substrate binding sites, it lowers the activity of all tested enzymes. PMID- 11856353 TI - Proteolytic action of duodenase is required to induce DNA synthesis in pulmonary artery fibroblasts. AB - Duodenase is a 29-kDa serine endopeptidase that displays selective trypsin- and chymotrypsin-like substrate specificity. This enzyme has been localized to epitheliocytes of Brunner's glands, and as described here, to mast cells within the intestinal mucosa and lungworm-infected lung, implying an important additional role in inflammation and tissue remodelling. In primary cultures of pulmonary artery fibroblasts, duodenase induced a concentration-dependent increase in [3H]thymidine incorporation with a maximal effect observed at 30 nm. Pretreating duodenase with soybean trypsin inhibitor abolished DNA synthesis, confirming that proteolytic activity was an essential requirement for this response. PAR1, PAR2 and PAR4 activating peptides were unable to induce [3H]thymidine incorporation in pulmonary artery fibroblasts. Likewise, pretreatment of fibroblasts with TNFalpha, known to up-regulate PAR2 expression in other systems, and IL-1beta, did not enhance the potential of duodenase to induce DNA synthesis. Furthermore, duodenase increased GTPgammaS binding to fibroblast membranes indicating that a G-protein-coupled receptor may mediate the effects of duodenase. Duodenase-induced DNA synthesis and GTPgammaS binding were both found to be inhibited by pertussis toxin, implying a role for Gi/o. Selective inhibitors of MEK1 (PD98059) and protein kinase C (GF109203X) only partially inhibited duodenase-induced DNA synthesis, but both wortmannin (100 nm) and LY294002 (10 microm) inhibited this response completely, indicating a key role for PtdIns 3-kinase. Furthermore, duodenase induced a 2.3 plus minus 0.1 fold increase in PtdIns 3-kinase activity in p85 immunoprecipitates, which was sensitive to inhibition by wortmannin. These results suggest that duodenase can induce pulmonary artery fibroblast DNA synthesis in a PtdIns 3-kinase-dependent manner via a G-protein-coupled receptor which is activated by a proteolytic mechanism. PMID- 11856352 TI - Repression of FasL expression by retinoic acid involves a novel mechanism of inhibition of transactivation function of the nuclear factors of activated T cells. AB - Retinoids are potent immune modulators that inhibit Fas ligand (FasL) expression and thereby repress the activation-induced apoptosis of immature thymocytes and T cell hybridomas. In this study, we demonstrate that all-trans-retinoic acid (all trans-RA) directly represses the transcriptional activity of the nuclear factors of activated T-cells (NFAT), which is an important transactivator of the FasL promoter. The analysis of reporter constructs containing the FasL promoter and wild-type or mutant NFAT binding-sites indicated that all-trans-RA repression was mediated via an NFAT binding element located in the promoter. A reporter construct comprising the NFAT binding sequence linked to a heterologous SV-40 promoter showed that NFAT transcriptional activity was significantly inhibited by all-trans-RA. Furthermore, all-trans-RA inhibited activation of the distal NFAT binding motif present in the interleukin (IL)-2 promoter, suggesting that the inhibition of NFAT function by all-trans-RA was not specific to the FasL promoter. Gel shift assays corroborated the results of the gene reporter studies by showing that all-trans-RA decreased the NFAT binding to DNA. All-trans-RA blocked translocation of NFATp from the cytosol into the nucleus, which was induced by PMA/ionomycin treatment in HeLa cells transfected with a Flag-tagged NFATp. Taken together, our results indicate that FasL inhibition by all-trans-RA involves a novel mechanism whereby the transcriptional function of NFAT is blocked. PMID- 11856355 TI - Structural and functional characterization of a C-type lectin-like antifreeze protein from rainbow smelt (Osmerus mordax). AB - Antifreeze proteins (AFPs) are produced by several cold-water fish species. They depress physiological freezing temperatures by inhibiting growth of ice crystals and, in so doing, permit the survival of these fish in seawater cooler than their normal freezing temperatures. The type II AFP from rainbow smelt (Osmerus mordax), which is a member of the C-type lectin superfamily, was characterized in terms of its Ca2+-binding quaternary structure and the role of its single N linked oligosaccharide. The protein core of the smelt AFP, shown through sequence homology to be a C-type lectin carbohydrate-recognition domain, was found to be protease resistant. Smelt AFP was also shown to bind Ca2+, as determined by ruthenium red staining and a conformational change on Ca2+ binding detected by intrinsic fluorescence. The N-linked oligosaccharide was found to have no effect on protease resistance, dimerization, or antifreeze activity. Thus its role, if any, in the antifreeze function of this protein remains unknown. Smelt AFP was also shown to be a true intermolecular dimer composed of two separate subunits. This dimerization did not require the presence of N-linked oligosaccharide or bound Ca2+. Smelt AFP dimerization has implications for the effective solution concentration and measurement of its activity. This finding may also lead to new interpretation of the mechanism of ice-growth inhibition by this AFP. PMID- 11856354 TI - Secretion of a peripheral membrane protein, MFG-E8, as a complex with membrane vesicles. AB - MFG-E8 (milk fat globule-EGF factor 8) is a peripheral membrane glycoprotein, which is expressed abundantly in lactating mammary glands and is secreted in association with fat globules. This protein consists of two-repeated EGF-like domains, a mucin-like domain and two-repeated discoidin-like domains (C-domains), and contains an integrin-binding motif (RGD sequence) in the EGF-like domain. To clarify the role of each domain on the peripheral association with the cell membrane, several domain-deletion mutants of MFG-E8 were expressed in COS-7 cells. The immunofluorescent staining of intracellular and cell-surface proteins and biochemical analyses of cell-surface-biotinylated and secreted proteins demonstrated that both of the two C-domains were required for the membrane association. During the course of these studies for domain functions, MFG-E8, but not C-domain deletion mutants, was shown to be secreted as membrane vesicle complexes. By size-exclusion chromatography and ultracentrifugation analyses, the complexes were characterized to have a high-molecular mass, low density and higher sedimentation velocity and to be detergent-sensitive. Not only such a exogenously expressed MFG-E8 but also that endogenously expressed in a mammary epithelial cell line, COMMA-1D, was secreted as the membrane vesicle-like complex. Scanning electron microscopic analyses revealed that MFG-E8 was secreted into the culture medium in association with small membrane vesicles with a size from 100 to 200 nm in diameter. Furthermore, the expression of MFG-E8 increased the number of these membrane vesicle secreted into the culture medium. These results suggest a possible role of MFG-E8 in the membrane vesicle secretion, such as budding or shedding of plasma membrane (microvesicles) and exocytosis of endocytic multivesicular bodies (exosomes). PMID- 11856356 TI - Novel regulatory regions found downstream of the rat B29/Ig-beta gene. AB - To search for novel regulatory regions, we examined the features of chromatin structure in the rat B29/Ig-beta gene and its flanking regions by determining DNase I hypersensitive sites (DHS) in plasmacytoma-derived Y3 cells. Six Y3 cell specific DHS were detected at -8.6, promoter, +0.7, +4.4, +6.0, and +8.7 kb. The DHS at +4.4, +6.0, and +8.7 kb were present in the intergenic region between B29/Ig-beta and growth hormone (GH) genes and were mapped inside conserved sequences in rat and humans. In transient transfection into Y3 cells, 2.9-kb DNA containing the +4.4 and +6.0-kb DHS demonstrated six times more enhancing activity than B29/Ig-beta promoter alone. Three intergenic DHS each possessed enhancing activity that was highest in the +4.4-kb region. In the electrophoretic mobility shift assay, a major band shift was demonstrated with Y3 nuclear extract and 0.3-kb DNA containing the +4.4-kb region with a conserved 0.22-kb sequence. By footprint analysis, 20 bases in the middle of the 0.3-kb DNA were protected by Y3 nuclear extract in which the consensus binding site for the OCT family was present. Deletion of the footprinted region reduced enhancing activity to that of the B29/Ig-beta promoter alone. The sequence responsible for the major band shift and transcriptional enhancing activity in the conserved +4.4-kb region thus coincided with the 20-bp footprinted region. PMID- 11856357 TI - Chemical structure and immunoreactivity of the lipopolysaccharide of the deep rough mutant I-69 Rd-/b+ of Haemophilus influenzae. AB - From the lipopolysaccharide of the deep rough mutant I-69 Rd--/b+ of Haemophilus influenzae two oligosaccharides were obtained after de-O-acylation and separation by high-performance anion exchange chromatography. Their chemical structures were determined by one- and two-dimensional 1H-, 13C- and 31P-NMR spectroscopy as alphaKdo-4P-(2-->6)-betaGlcN-4P-(1-->6)-alphaGlcN-1P and alphaKdo-5P-(2-->6) betaGlcN-4P-(1-->6)-alphaGlcN-1P. The specificity of mAbs S42-21 and S42-16 specific for Kdo-4P or Kdo-5P, respectively [Rozalski, A., Brade L., Kosma P., Moxon R., Kusumoto S., & Brade H. (1997). Mol. Microbiol. 23, 569--577] was confirmed with neoglycoconjugates obtained by conjugation of the isolated oligosaccharides to BSA. In addition, a mAb S42-10-8 with unknown epitope specificity could be assigned using the neoglycoconjugates described herein. This mAb binds to an epitope composed of the bisphosphorylated glucosamine backbone of lipid A and Kdo-4P, whereby the latter determines the specificity strictly by the position of the phosphate group. PMID- 11856358 TI - Exploration of the diaphorase activity of neutrophil NADPH oxidase. AB - In the O2- generating flavocytochrome b, the membrane-bound component of the neutrophil NADPH oxidase, electrons are transported from NADPH to O2 in the following sequence: NADPH --> FAD --> heme b -->O2. Although p iodonitrotetrazolium (INT) has frequently been used as a probe of the diaphorase activity of the neutrophil flavocytochrome b, the propensity of its radical to interact reversibly with O2 led us to question its specificity. This study was undertaken to reexamine the interaction of INT with the redox components of the neutrophil flavocytochrome b. Two series of inhibitors were used, namely the flavin analog 5-deaza FAD and the heme inhibitors bipyridyl and benzylimidazole. The following results indicate that INT reacts preferentially with the hemes rather than with the FAD redox center of flavocytochrome b and is not therefore a specific probe of the diaphorase activity of flavocytochrome b. First, in anaerobiosis, reduced heme b in activated membranes was reoxidized by INT as efficiently as by O2 even in the presence of concentrations of 5-deaza FAD which fully inhibited the NADPH oxidase activity. Second, the titration curve of dithionite-reduced heme b in neutrophil membranes obtained by oxidation with increasing amounts of INT was strictly superimposable on that of dithionite reduced hemin. Third, INT competitively inhibited the O2 uptake by the activated NADPH oxidase in a cell-free system. Finally, the heme inhibitor bipyridyl competitively inhibited the reduction of INT in anaerobiosis, and the oxygen uptake in aerobiosis. PMID- 11856359 TI - The DNA-polymerase inhibiting activity of poly(beta-l-malic acid) in nuclear extract during the cell cycle of Physarum polycephalum. AB - The naturally synchronous plasmodia of myxomycetes synthesize poly(beta-l-malic acid), which carries out cell-specific functions. In Physarum polycephalum, poly(beta-l-malate) [the salt form of poly(beta-l-malic acid)] is highly concentrated in the nuclei, repressing DNA synthetic activity of DNA polymerases by the formation of reversible complexes. To test whether this inhibitory activity is cell-cycle-dependent, purified DNA polymerase alpha of P. polycephalum was added to the nuclear extract and the activity was measured by the incorporation of [3H]thymidine 5'-monophosphate into acid precipitable nick activated salmon testis DNA. Maximum DNA synthesis by the reporter was measured in S-phase, equivalent to a minimum of inhibitory activity. To test for the activity of endogenous DNA polymerases, DNA synthesis was followed by the highly sensitive photoaffinity labeling technique. Labeling was observed in S-phase in agreement with the minimum of the inhibitory activity. The activity was constant throughout the cell cycle when the inhibition was neutralized by the addition of spermidine hydrochloride. Also, the concentration of poly(beta-l-malate) did not vary with the phase of the cell cycle [Schmidt, A., Windisch, C. & Holler, E. (1996) Nuclear accumulation and homeostasis of the unusual polymer poly(beta-l malate) in plasmodia of Physarum polycephalum. Eur. J. Cell Biol. 70, 373-380]. To explain the variation in the cell cycle, a periodic competition for poly(beta l-malate) between DNA polymerases and most likely certain histones was assumed. These effectors are synthesized in S-phase. By competition they displace DNA polymerase from the complex of poly(beta-l-malate). The free polymerases, which are no longer inhibited, engage in DNA synthesis. It is speculated that poly(beta l-malate) is active in maintaining mitotic synchrony of plasmodia by playing the mediator between the periodic synthesis of certain proteins and the catalytic competence of DNA polymerases. PMID- 11856360 TI - Solution structure of a hydrophobic analogue of the winter flounder antifreeze protein. AB - The solution structure of a synthetic mutant type I antifreeze protein (AFP I) was determined in aqueous solution at pH 7.0 using nuclear magnetic resonance (NMR) spectroscopy. The mutations comprised the replacement of the four Thr residues by Val and the introduction of two additional Lys-Glu salt bridges. The antifreeze activity of this mutant peptide, VVVV2KE, has been previously shown to be similar to that of the wild type protein, HPLC6 (defined here as TTTT). The solution structure reveals an alphahelix bent in the same direction as the more bent conformer of the published crystal structure of TTTT, while the side chain chi1 rotamers of VVVV2KE are similar to those of the straighter conformer in the crystal of TTTT. The Val side chains of VVVV2KE assume the same orientations as the Thr side chains of TTTT, confirming the conservative nature of this mutation. The combined data suggest that AFP I undergoes an equilibrium between straight and bent helices in solution, combined with independent equilibria between different side chain rotamers for some of the amino acid residues. The present study presents the first complete sequence-specific resonance assignments and the first complete solution structure determination by NMR of any AFP I protein. PMID- 11856361 TI - Kinetic properties of bifunctional 6-phosphofructo-2-kinase/fructose-2,6 bisphosphatase from spinach leaves. AB - A cDNA encoding 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase was isolated from a Spinacia oleracea leaf library and used to express a recombinant enzyme in Escherichia coli and Spodoptera frugiperda cells. The insoluble protein expressed in E. coli was purified and used to raise antibodies. Western blot analysis of a protein extract from spinach leaf showed a single band of 90.8 kDa. Soluble protein was purified to homogeneity from S. frugiperda cells infected with recombinant baculovirus harboring the isolated cDNA. The soluble protein had a molecular mass of 320 kDa, estimated by gel filtration chromatography, and a subunit size of 90.8 kDa. The purified protein had activity of both 6 phosphofructo-2-kinase specific activity 10.4-15.9 nmol min(-1) x mg protein (-1) and fructose-2,6-bisphosphatase (specific activity 1.65-1.75 nmol x mol(-1) mg protein(-1). The 6-phosphofructo-2-kinase activity was activated by inorganic phosphate, and inhibited by 3-carbon phosphorylated metabolites and pyrophosphate. In the presence of phosphate, 3-phosphoglycerate was a mixed inhibitor with respect to both fructose 6-phosphate and ATP. Fructose-2,6 bisphosphatase activity was sensitive to product inhibition; inhibition by inorganic phosphate was uncompetitive, whereas inhibition by fructose 6-phosphate was mixed. These kinetic properties support the view that the level of fructose 2,6-bisphosphate in leaves is determined by the relative concentrations of hexose phosphates, three-carbon phosphate esters and inorganic phosphate in the cytosol through reciprocal modulation of 6-phosphofructo-2-kinase and fructose-2,6 bisphosphatase activities of the bifunctional enzyme. PMID- 11856362 TI - Purification and characterization of the thyrotropin-releasing hormone (TRH) degrading serum enzyme and its identification as a product of liver origin. AB - Previous biochemical studies have indicated that the membrane-bound thyrotropin releasing hormone (TRH)-degrading enzyme (TRH-DE) from brain and liver and the serum TRH-DE are derived from the same gene. These studies also suggested that the serum enzyme is of liver origin. The present study was undertaken to verify these hypotheses. In different species, a close relationship between the activities of the serum enzyme and the particulate liver enzyme was noticed. The activity of the serum enzyme decreased when rats were treated with thioacetamide, a known hepatotoxin. With hepatocytes cultured in a sandwich configuration, release of the TRH-DE into the culture medium could also be demonstrated. The trypsin-solubilized particulate liver TRH-DE and the serum TRH-DE were purified to electrophoretic homogeneity. Both enzymes and the brain TRH-DE were recognized by a monoclonal antibody generated with the purified brain enzyme as antigen. Lectin blot analysis indicated that the serum enzyme and the liver enzyme are glycoproteins containing a sugar structure of the complex type, whereas the brain enzyme exhibits an oligomannose/hybrid glycostructure. A molecular mass of 97 000 Da could be estimated for all three enzymes after deglycosylation and SDS/PAGE followed by Western blotting. Fragment analysis of the serum TRH-DE revealed that the peptide sequences correspond to the cDNA deduced amino-acid sequences of the membrane-bound brain TRH-DE, whereby two peptides were identified that are encoded by exon 1. These data strongly support the hypothesis that the TRH-DEs are all derived from the same gene, whereby the serum enzyme is generated by proteolytic cleavage of the particulate liver enzyme. PMID- 11856363 TI - Inactivation of the Na+-translocating NADH:ubiquinone oxidoreductase from Vibrio alginolyticus by reactive oxygen species. AB - The Na+-translocating NADH:quinone oxidoreductase (Na+-NQR) from Vibrio alginolyticus was inactivated by reactive oxygen species. Highest Na+-NQR activity was observed in anaerobically prepared membranes that exhibited 1:1 coupling of NADH oxidation and Q reduction activities (1.6 U x mg(-1)). Optical and EPR spectroscopy documented the presence of b-type cytochromes, a [2Fe-2S] cluster and an organic radical signal in anaerobically prepared membranes from V. alginolyticus. It is shown that the [2Fe-2S] cluster previously assigned to the Na+-NQR originates from the succinate dehydrogenase or the related enzyme fumarate reductase. PMID- 11856364 TI - Interallelic recombination is probably responsible for the occurrence of a new alpha(s1)-casein variant found in the goat species. AB - The alphas1-casein (alphas1-Cas) locus in the goat is characterized by a polymorphism, the main feature of which is to be qualitative as well as quantitative. A systematic analysis performed in an autochthon southern Italy breed identified a new rare allele (M), which was characterized at both the protein and genomic level. The M protein displays the slowest electrophoretic mobility of the alphas1-Cas variants described so far. MS and automated Edman degradation experiments showed that this behavior was due to the loss of two phosphate residues in the multiple phosphorylation site (64SP-SP-SP-SP-SP-E-70E) consecutively to a Ser-->Leu substitution at position 66 of the peptide chain (64S-SP-L-SP-SP-E-70E). This was confirmed by sequencing a genomic DNA fragment encompassing exon 9 where the 8th codon (TCG) was shown to be mutated to TTG. Sequencing of amplified genomic DNA segments spanning the 5' and 3' flanking regions of each exon allowed us to identify 23 single nucleotide polymorphisms and two insertion/deletion events in the coding as well as the noncoding regions. A comparison of specific haplotypes defined for each of the alphas1-CasF, A and M alleles indicates that the M allele probably arises from interallelic recombination between alleles A and B2, followed by a C-->T transition at nucleotide 23 of the ninth exon. The region encompassing the recombination break point was putatively located between nucleotide 86 upstream and nucleotide 40 downstream of exon 8. Interallelic recombination therefore appears to be a possible means of generating allelic diversity at the alphas1-Cas locus, at least in the goat. The previously proposed molecular phylogeny must now be revised, possibly starting from two ancestral allelic lineages. PMID- 11856365 TI - Divergent members of a soybean (Glycine max L.) 4-coumarate:coenzyme A ligase gene family. AB - 4-Coumarate:CoA ligase (4CL) is involved in the formation of coenzyme A thioesters of hydroxycinnamic acids that are central substrates for subsequent condensation, reduction, and transfer reactions in the biosynthesis of plant phenylpropanoids. Previous studies of 4CL appear to suggest that many isoenzymes are functionally equivalent in supplying substrates to various subsequent branches of phenylpropanoid biosyntheses. In contrast, divergent members of a 4CL gene family were identified in soybean (Glycine max L.). We isolated three structurally and functionally distinct 4CL cDNAs encoding 4CL1, 4CL2, and 4CL3 and the gene Gm4CL3. A fourth cDNA encoding 4CL4 had high similarity with 4CL3. The recombinant proteins expressed in Escherichia coli possessed highly divergent catalytic efficiency with various hydroxycinnamic acids. Remarkably, one isoenzyme (4CL1) was able to convert sinapate; thus the first cDNA encoding a 4CL that accepts highly substituted cinnamic acids is available for further studies on branches of phenylpropanoid metabolism that probably lead to the precursors of lignin. Surprisingly, the activity levels of the four isoenzymes and steady-state levels of their transcripts were differently affected after elicitor treatment of soybean cell cultures with a beta-glucan elicitor of Phytophthora sojae, revealing the down-regulation of 4CL1 vs. up-regulation of 4CL3/4. A similar regulation of the transcript levels of the different 4CL isoforms was observed in soybean seedlings after infection with Phytophthora sojae zoospores. Thus, partitioning of cinnamic acid building units between phenylpropanoid branch pathways in soybean could be regulated at the level of catalytic specificity and the level of expression of the 4CL isoenzymes. PMID- 11856366 TI - Ornithine decarboxylase-antizyme is rapidly degraded through a mechanism that requires functional ubiquitin-dependent proteolytic activity. AB - Antizyme is a polyamine-induced cellular protein that binds to ornithine decarboxylase (ODC), and targets it to rapid ubiquitin-independent degradation by the 26S proteasome. However, the metabolic fate of antizyme is not clear. We have tested the stability of antizyme in mammalian cells. In contrast with previous studies demonstrating stability in vitro in a reticulocyte lysate-based degradation system, in cells antizyme is rapidly degraded and this degradation is inhibited by specific proteasome inhibitors. While the degradation of ODC is stimulated by the presence of cotransfected antizyme, degradation of antizyme seems to be independent of ODC, suggesting that antizyme degradation does not occur while presenting ODC to the 26S proteasome. Interestingly, both species of antizyme, which represent initiation at two in-frame initiation codons, are rapidly degraded. The degradation of both antizyme proteins is inhibited in ts20 cells containing a thermosensitive ubiquitin-activating enzyme, E1. Therefore we conclude that in contrast with ubiquitin-independent degradation of ODC, degradation of antizyme requires a functional ubiquitin system. PMID- 11856367 TI - Short peptides are not reliable models of thermodynamic and kinetic properties of the N-terminal metal binding site in serum albumin. AB - A comparative study of thermodynamic and kinetic aspects of Cu(II) and Ni(II) binding at the N-terminal binding site of human and bovine serum albumins (HSA and BSA, respectively) and short peptide analogues was performed using potentiometry and spectroscopic techniques. It was found that while qualitative aspects of interaction (spectra and structures of complexes, order of reactions) could be reproduced, the quantitative parameters (stability and rate constants) could not. The N-terminal site in HSA is much more similar to BSA than to short peptides reproducing the HSA sequence. A very strong influence of phosphate ions on the kinetics of Ni(II) interaction was found. This study demonstrates the limitations of short peptide modelling of Cu(II) and Ni(II) transport by albumins. PMID- 11856368 TI - Movement of yeast 1,3-beta-glucan synthase is essential for uniform cell wall synthesis. AB - BACKGROUND: The cell wall has an important role in maintaining cell shape. In the budding yeast Saccharomyces cerevisiae, the major filamentous component of the cell wall responsible for its rigidity is 1,3-beta-glucan and is synthesized by 1,3-beta-glucan synthase (GS), localized on the plasma membrane. RESULTS: Observations of green fluorescent protein (GFP)-conjugated Fks1p, a catalytic subunit of GS, revealed that it is co-localized with cortical actin patches and moves on the cell surface at the sites of cell wall remodelling. Mutants with impaired actin patch movement show immobility of Fks1p-GFP spots, indicating that actin patch motility is required for the movement of Fks1p. Cells with immobilized Fks1p exhibit defective cell wall structure and function. The cell wall thickness of the mutants becomes irregular, eventually leading to cell lysis. CONCLUSION: We propose that GS movement is necessary for proper cell wall remodelling. PMID- 11856369 TI - Aurora-B phosphorylates Histone H3 at serine28 with regard to the mitotic chromosome condensation. AB - BACKGROUND: Histone H3 (H3) phosphorylation plays important roles in mitotic chromosome condensation. We reported that H3 phosphorylation occurs at Ser28, as well as at Ser10 during mitosis, at least in mammals. Aurora B was recently demonstrated to be responsible for Ser10 phosphorylation in S. cerevisiae, C. elegans, Drosophila and Xenopus egg extract. RESULTS: We compared the distribution of Aurora-B with that of H3 phosphorylation. Aurora-B was primarily localized in the heterochromatin of late G2 phase cells, where only Ser10 phosphorylation was observed. The treatment of such cells with calyculin A induced Ser28 phosphorylation in the Aurora-B-localized area. During prophase to metaphase, Aurora-B was distributed in condensing chromosomes where Ser10 and Ser28 were phosphorylated. Aurora-B can phosphorylate H3-Ser10 and -Ser28 in nucleosomes in vitro. Transfection of a dominant-negative mutant of Aurora-B resulted in a reduction of H3 phosphorylation, not only at Ser10 but also Ser28, during mitosis. CONCLUSIONS: With regard to mitotic chromosome condensation, Aurora-B directly phosphorylated H3, not only at Ser10 but also at Ser28. The level of Ser28 phosphorylation is diminished to undetectable levels by PP1 phosphatase prior to entry into mitosis. PMID- 11856370 TI - Coaction of DNA topoisomerase IIIalpha and a RecQ homologue during the germ-line mitosis in Caenorhabditis elegans. AB - BACKGROUND: Among the four RecQ homologues predicted from the Caenorhabditis elegans genomic DNA sequence, T04A11.6 is most similar to Bloom syndrome's protein in humans. To investigate a possible interaction of the protein with topoisomerase IIIalpha (TOP3alpha), as observed between TOP3 and RecQ homologues in yeast and human, the top3alpha gene expression was suppressed by RNA interference (RNAi) in the him-6(e1104) C. elegans strain which is mutated in T04A11.6 (F. Mueller & C. Wicky, personal communication). RESULTS: Germ cells in the gonads of the progeny him-6(e1104);top3alpha(RNAi) showed severe chromosomal abnormalities and were arrested during mitosis with a subsequent failure in meiotic entry. Most of the aberrant chromosomes were stained by the TUNEL assay but not by the SYTO12 dye, suggesting extensive DNA breaks not associated with apoptosis. The phenotypes in the germ cells of him-6(e1104);top3alpha(RNAi) were also observed in the progeny produced by double RNA interference of the top3alpha and him-6 gene expression, though at a reduced level. The over-expressed TOP3alpha and Him-6 proteins showed specific physical interaction in vitro, in agreement with the genetic interaction in C. elegans. CONCLUSION: In C. elegans, TOP3alpha and the RecQ homologue (T04A11.6) contribute to genome stability during germ-line mitosis, probably by acting in a complex. PMID- 11856371 TI - Interaction of PRMT1 with BTG/TOB proteins in cell signalling: molecular analysis and functional aspects. AB - BACKGROUND: Several recent reports have connected protein methylation with differentiation. Furthermore, the BTG/TOB proteins have also been implicated in such control. BTG1 and 2 have been shown to interact with PRMT1 (predominant cellular arginine N-methyltransferase of type I). RESULTS: First, we have studied the interaction between PRMT1 and the proteins of the BTG/TOB family. We show that boxC, a sequence present only in BTG1 and BTG2, is essential for this association. Using boxC peptide, we have investigated the importance of PRMT1/BTG protein association during type I protein methylation reactions. Finally, we show that the addition of boxC fused to penetratin interferes with the neuronal differentiation of PC12 cells and ES cell-derived neurones. CONCLUSIONS: Taken together, these results indicate that PRMT1/BTG proteins could play a key role in the arginine methylation-mediated signalling pathway as well as in neuronal differentiation. PMID- 11856372 TI - Essential role of S-adenosylmethionine decarboxylase in mouse embryonic development. AB - BACKGROUND: S-Adenosylmethionine decarboxylase (AdoMetDC) is one of the key enzymes involved in the biosynthesis of spermidine and spermine, which are essential for normal cell growth. To examine the role of polyamines in embryogenesis, we carried out targeted disruption of the mouse Amd1 gene, encoding AdoMetDC, to generate mice that can not synthesize spermidine and spermine. RESULTS: Amd1 heterozygous mice were viable, normal and fertile. However, homozygous Amd1(-/-) embryos died early in embryonic development, between E3.5 and E6.5 days post-coitus. Homozygous (Amd1(-/-)) blastocysts at E3.5 arrested cell proliferation immediately after the onset of cell culture, and this arrest was rescued by the addition of spermidine. Chromosomal DNA breakage did not occur in Amd1(-/-) blastocysts at E3.5, as determined by TUNEL assay. CONCLUSIONS: These results indicate that AdoMetDC plays an essential role in embryonic development and that polyamines are required for cell proliferation in the embryo after E3.5. PMID- 11856373 TI - The H1 and H2 regions of the activation domain of herpes simplex virion protein 16 stimulate transcription through distinct molecular mechanisms. AB - BACKGROUND: The Herpes Simplex Virion Protein 16 (VP16) contains a strong activation domain which can be subdivided into two regions, H1 and H2, both of which independently activate transcription in vivo. Several components of the basal transcription machinery have been shown to interact with the activation domain of VP16, mostly through the H1 region. RESULTS: We show that the H2 region binds directly to histone acetyltransferase, CBP (CREB (cAMP Responsive Element Binding Protein) Binding Protein) both in vivo and in vitro. The sites of interaction with the H2 region were mapped to both the amino- and carboxy terminal segments of CBP. A mutation in the H2 region disrupts the interaction with CBP and abolishes the ability of VP16 to mediate in vitro transactivation from chromatin templates in an acetyl-CoA dependent manner. In contrast, human Mediator, another co-activator complex, binds specifically to both the H1 and H2 regions. CONCLUSION: The H1 and H2 regions of the VP16 activation domain activate transcription via distinct pathways. The H2 requires CBP for activation, whereas the H1 may function through Mediator and general transcription factors. PMID- 11856374 TI - Polyanionic stretch-deleted histone chaperone cia1/Asf1p is functional both in vivo and in vitro. AB - BACKGROUND: CIA, an interactor of the CCG1 histone acetyltransferase subunit of TFIID, was identified as a human histone chaperone. The Saccharomyces cerevisiae orthologue ASF1, when it was over-expressed, was reported to cause de-repression of silent loci; however, the involvement of Asf1p in the alteration of nucleosomal structures remained unknown. Curiously, there is a polyanionic stretch, a structural motif characteristic of histone chaperones, in S. cerevisiae Asf1p, but not in human CIA. We investigated how CIA/Asf1p utilizes its domain(s) for the alteration of nucleosomal structure. RESULTS: To characterize the relationships between the domain structures and nuclear functions of CIA, we isolated the gene for the CIA counterpart in Schizosaccharomyces pombe, designated cia1+, whose putative product contains a polyanionic stretch. Gene disruption of cia1+ was lethal, which is the distinct phenotype of viable S. cerevisiae asf1. The cia1- lethality was rescued by the introduction of S. cerevisiae ASF1, but not by the introduction of human CIA cDNA. To our surprise, the construct that produces Asf1p, lacking the polyanionic stretch, is capable of rescuing the lethality caused by the cia1+ deletion, while the highly conserved N-terminal region of Asf1p is essential for the complementation of cia1- growth defects. The polyanionic stretch-deleted Asf1p is sufficient both for interaction with histones H3/H4 and for nucleosome assembly in vitro, as well as for telomeric de-repression in vivo. CONCLUSION: These findings suggest that the areas responsible for both the conserved and species specific functions of CIA/cia1/Asf1p are within their highly conserved regions and that the yeast-specific polyanionic stretch of cia1/Asf1p is not necessary for viability, histone binding, nucleosome assembly, or anti-silencing. PMID- 11856375 TI - Paz2 and 13 other PAZ gene products regulate vacuolar engulfment of peroxisomes during micropexophagy. AB - BACKGROUND: In the methylotrophic yeast Pichia pastoris, peroxisomes can be selectively degraded through direct engulfment by the vacuole in a process known as micropexophagy, but the mechanism of micropexophagy is not known. RESULTS: To gain molecular insights into micropexophagy, we used fluorescence time-lapse microscopy, coupled with gene-tagging mutagenesis to isolate P. pastoris mutants defective in micropexophagy. The relevant genes have been designated PAZ genes. Morphological and genetic analyses enabled us to postulate a schematic model for micropexophagy. This new model invokes the generation of new vacuolar compartments as an intermediate structure during micropexophagy. Different classes of paz mutants arrest micropexophagy at distinct stages of the process. Most of APG-related paz mutants ceased micropexophagy at Stage 1c and that GCN family paz mutants ceased micropexophagy at Stage 2. The paz2Delta strain shows a unique phenotype. Paz2 is the homologue of Saccharomyces cerevisiae Apg8, which is necessary for macroautophagy in that yeast. Our analysis revealed that in P. pastoris, Paz2 plays a key role in repressing the engulfment of peroxisomes by the vacuole before the onset of micropexophagy. Paz2 is proteolytically processed by another autophagy-related Paz protein Paz8, but this processing is not required for the ability of Paz2 to suppress aberrant micropexophagy. CONCLUSION: Micropexophagy has been dissected into a multistep reaction that involves 14 identified Paz gene products. Our studies indicate that Paz2 controls the engulfment of peroxisomes by the vacuole, pointing to a novel early function of this protein. PMID- 11856376 TI - Nanosecond fluorescence resonance energy transfer-fluorescence lifetime imaging microscopy to localize the protein interactions in a single living cell. AB - Visualizing and quantifying protein-protein interactions is a recent trend in biomedical imaging. The current advances in fluorescence microscopy, coupled with the development of new fluorescent probes such as green fluorescent proteins, allow fluorescence resonance energy transfer (FRET) to be used to study protein interactions in living specimens. Intensity-based FRET microscopy is limited by spectral bleed-through and fluorophore concentration. Fluorescence lifetime imaging (FLIM) microscopy and lifetime measurements are independent of change in fluorophore concentration or excitation intensity, and the combination of FRET and FLIM provides high spatial (nanometre) and temporal (nanoseconds) resolution. Because only the donor fluorophore lifetime is measured, spectral bleed-through is not an issue in FRET-FLIM imaging. In this paper we describe the development of a nanosecond FRET-FLIM microscopy instrumentation to acquire the time-resolved images of donor in the presence and the absence of the acceptor. Software was developed to process the acquired images for single and double exponential decays. Measurement of donor lifetime in two different conditions allowed us to calculate accurately the distance between the interacting proteins. We used this approach to quantify the dimerization of the transcription factor CAATT/enhancer binding protein alpha in living pituitary cells. The one- and two-component analysis of the donor molecule lifetime in the presence of acceptor demonstrates the distance distribution between interacting proteins. PMID- 11856377 TI - Shearing microscopy using polarized optical microscope with shear stage and spectral analyser to study liquid crystalline polymers. AB - In-situ polarized optical microscopy using a shear stage and a spectral analyser as well as a CCD camera were applied to study the phase transition under shear flow for a thermotropic and side-chain-type liquid crystalline polysiloxane. The onset of the appearance of anisotropic texture of the polysiloxane was observed under shear flow using the CCD camera at temperatures much higher than the isotropic-liquid crystalline phase transition temperature if the polysiloxane was cooled from the isotropic phase in the quiescent state. Both the onset temperature and the temperature for full development of the anisotropic texture across the field of view became higher as the shear rate increased. The transmitted light intensity was also measured using a spectral analyser with crossed polarisers at wavelengths from 300 nm to 800 nm, and the integrated intensity of the spectrum was calculated. Changes in the spectrum and the integrated intensity against temperature in the cooling process were compared with observation using the CCD camera. Temperature dependence of the integrated intensity showed that the onset of the appearance of the anisotropic texture under high shear rates was detected at temperatures slightly higher than that observed using the CCD camera. PMID- 11856378 TI - Study of carbon fibres and carbon-carbon composites by scanning thermal microscopy. AB - Scanning thermal microscopy (SThM) is a relatively new technique based on atomic force microscopy in which the tip is replaced by an ultra-miniature temperature probe. This paper reports on a preliminary investigation of the application of SThM in the characterization of the thermal properties of carbon fibres and carbon-carbon (CC) composites. The technique enabled a comparative study to be made of discrete fibre and matrix thermal properties in a series of model unidirectional composites. The thermal images revealed a marked increase in thermal conductivity of the matrix with increasing temperature of treatment and hence confirmed the development of a highly ordered carbon matrix. The results were in qualitative agreement with previously determined values of thermal conductivity from which the separate values of fibre and matrix thermal conductivity had been derived. The technique was also applied to the characterization of samples of unknown processing history, enabling an estimation to be made of the heat treatment and type of the fibres and matrix present in the composite. It was concluded that SThM promises to be a powerful technique for the study of the thermal properties of CC composites and carbon fibres, as it uniquely enables variations in local thermal conductivity to be detected and resolved. Absolute quantification of the technique remains the key to its future widespread acceptance in materials characterization. PMID- 11856379 TI - A metadata classification schema for semantic content analysis of videos. AB - Simple ancillary metadata, such as those encompassed by the 15 elements of the Dublin Core, may be sufficient and entirely appropriate for basic coarse granularity cross-domain resource discovery. However, they are insufficient and inappropriate for content description of complex data types such as videos, which require more detailed relational models. We propose a metadata classification schema for the characterization of items and events in videos that permits subsequent query by content. Following MPEG-7 nomenclature, metadata intrinsic to the information content of the video are defined as either structural or semantic, where structural metadata are numerical feature primitives produced by analysing the colour, shape, texture, structure and motion within the video frames, whereas semantic metadata describe the locations and timings of individual items and particular actions or events in the video, and are thus of higher information value. In this paper, the semantic metadata required to describe the visual information content of videos are defined and classified into four distinct classes: Media Entities; Content Items; Events; and Supplementary Items, and three types of property tables are defined: Identity Tables; Spatio Temporal Position Tables; and Event Tables, in which these metadata may be stored in a relational database. PMID- 11856380 TI - VANQUIS, a system for the interactive semantic content analysis and spatio temporal query by content of videos. AB - Using the video metadata descriptors and data model defined in the accompanying paper (Shotton, D. M. et al. (2002) A metadata classification schema for semantic content analysis of videos. J. Microsc. 205, 33-42), we discuss how analysis of the content of scientific videos, and subsequent query by content of the resulting semantic metadata, can be enhanced by the use of an object-relational database. We illustrate this by describing VANQUIS, a Web-based prototype video analysis and query interface system for the interactive spatio-temporal analysis and subsequent query by content of videos. Using VANQUIS to generate standard SQL (structured query language) statements that address complex data types stored in an object-relational database, relationships between characters and events contained within and between videos can be identified, and the appropriate video segments containing these characters and events can be retrieved for viewing. We give examples of analysis and query implementation by using VANQUIS to analyse a biological microscopy video, and discuss the wider potential of this methodology for the analysis and query by content of videos containing more general subject matter. PMID- 11856381 TI - Near field optical microscopy in aqueous solution: implementation and characterization of a vibrating probe. AB - Near field optical microscopy (NSOM) is one of the possible solutions to circumvent the diffraction limit, but the control of the optical probe in solution has been a technical challenge for practical applications. Most recently, it has been shown that the pipette used in the scanning ion conductance microscope can be modified to form a high resolution near field optical probe. When combined with a novel distance modulation mechanism, a robust near field microscope can be constructed for operation in aqueous solution. In this paper, we present technical details of this design and a further characterization of the NSOM system for imaging in solution. Fundamental limitations of this approach in comparison to other systems are also discussed. Based on the current technology, it is concluded that better than 50 nm resolution should be achievable with this technique for fluorescence, as well as fluorescence resonance energy transfer, imaging of biological specimens. PMID- 11856383 TI - Optical far-field microscopy of single molecules with 3.4 nm lateral resolution. AB - Optical far-field imaging of single molecules in a frozen solution at 1.2 K with a lateral resolution of 3.4 nm is reported. The mechanical stability of the fluorescence microscope, especially of the low-temperature insert, allows for the localization of fluorescing molecules with a reproducibility of better than 5 nm within observation times up to 10 min. For observation times of 9 h the reproducibility of the lateral position is limited to about 20 nm due to mechanical drift. Lateral position and orientation of 314 single molecules, present within the confocal detection volume of approximately 10 microm(3), are obtained. The possibility to correct for mechanical drift by monitoring the position of a spatial reference in the sample is demonstrated. PMID- 11856382 TI - Measurement of the three-dimensional microscope point spread function using a Shack-Hartmann wavefront sensor. AB - We present a technique to measure the wavefront in the exit pupil of a microscope to determine the microscope's three-dimensional point spread function (PSF) experimentally. The wavefront yields the microscope PSF through a Fourier transform that models propagation of light from the exit pupil to the image plane. A Shack-Hartmann wavefront sensor is used to measure the wavefront shape by recording lateral displacements of a grid of focused spots created by a lenslet array. The displacement of each spot is related to the local wavefront slope. Thus, with appropriate sampling across the exit pupil, the entire wavefront can be reconstructed. This technique does not require the use of a sub resolution object to obtain the three-dimensional microscope PSF. Consequently, larger, brighter fluorescent objects may be imaged, thereby reducing the requirements for detector sensitivity and leading to a three-fold increase in the axial range over which the PSF is measured. The Shack-Hartmann technique results in a description of the PSF as a continuous function whose sampling is not dependent on the size of the CCD pixels. The Shack-Hartmann sensor is not limited by the numerical aperture of the objective and can easily be calibrated to measure the PSF at any wavelength. PMID- 11856384 TI - On the role of electron-ion recombination in low vacuum scanning electron microscopy. AB - Here we demonstrate the effects of electron-ion recombination on imaging signals utilized in low vacuum scanning electron microscopes (SEMs). The presented results show that, under normal operating conditions, recombination of ionized gas molecules with secondary electrons (SEs) suppresses a significant fraction of emitted electrons. If the ion flux (and hence the spatial dependence of the SE ion recombination rate) is laterally inhomogeneous across the imaged region of a specimen, contrast in SE images can be influenced and in some cases (under conditions of high detector field strength and long ionic mean free path) dominated by variations in the recombination rate. Consequently, SE images of features such as topographic asperities can exhibit edge-darkening, leading to inversion of some topographic contrast. Recognition of the extent and nature of electron-ion recombination is required for a correct understanding of processes occurring in variable pressure SEMs and, subsequently, for models of image formation. PMID- 11856385 TI - Consequences of positive ions upon imaging in low vacuum scanning electron microscopy. AB - The effects caused by an excess quantity of ionized gas molecules within the low vacuum, variable pressure and environmental scanning electron microscope (ESEM) are described with reference to mechanisms by which they can influence imaging conditions. These effects can include specimen charging, recombination and development of space charge. They are demonstrated for three different classes of sample: (1) an electrically grounded conductor, (2) an electrically floating conductor, and (3) an electrical insulator. A new device is presented that will aid excess charge removal within the ESEM and help correct for some of these effects, thereby dramatically improving imaging over a wide range of operating conditions and samples. The mechanism of image enhancement is demonstrated with reference to the three classes of sample described above. PMID- 11856386 TI - Comparative study of a block copolymer morphology by transmission electron microscopy and scanning force microscopy. AB - Using transmission electron microscopy (TEM) and scanning force microscopy (SFM) together, it was possible to verify important structural features of a nanostructured bulk material such as the kp-morphology in an ABC triblock copolymer. By applying suitable imaging techniques during the SFM measurements it was possible to determine the morphology without additional manipulation steps in between. In comparison, TEM investigations on this type of material usually require selective staining procedures prior to the measurement. Also electron beam damage is often encountered during TEM measurements especially if components such as poly(methacrylates) are present. In contrast, SFM measurements can be assumed not to significantly change the phase dimensions of the components. PMID- 11856387 TI - Fluorescence localization after photobleaching (FLAP): a new method for studying protein dynamics in living cells. AB - FLAP is a new method for localized photo-labelling and subsequent tracking of specific molecules within living cells. It is simple in principle, easy to implement and has a wide potential application. The molecule to be located carries two fluorophores: one to be photobleached and the other to act as a reference label. Unlike the related methods of fluorescence recovery after photobleaching (FRAP) and fluorescence loss in photobleaching (FLIP), the use of a reference fluorophore permits the distribution of the photo-labelled molecules themselves to be tracked by simple image differencing. In effect, FLAP is therefore comparable with methods of photoactivation. Its chief advantage over the method of caged fluorescent probes is that it can be used to track chimaeric fluorescent proteins directly expressed by the cells. Although methods are being developed to track fluorescent proteins by direct photoactivation, these still have serious drawbacks. In order to demonstrate FLAP, we have used nuclear microinjection of cDNA fusion constructs of beta-actin with yellow (YFP) and cyan (CFP) fluorescent proteins to follow both the fast relocation dynamics of monomeric (globular) G-actin and the much slower dynamics of filamentous F-actin simultaneously in living cells. PMID- 11856388 TI - Five years of aftercare of implant-retained mandibular overdentures and conventional dentures. AB - The purpose of this multicentre randomized clinical trial was to analyse surgical and prosthetic aftercare and clinical implant performance of edentulous patients with implant-retained mandibular overdentures and of patients with conventional dentures, either or not after pre-prosthetic vestibuloplasty and deepening of the floor of the mouth. The evaluation period was 5 years. The implant systems evaluated were the IMZ implant system, the Branemark implant system and the Transmandibular Implant system. The centre in Groningen had five groups (n=149) and the centre in Nijmegen had three groups (n=86). The evaluation comprised of surgical and prosthetic aftercare, together with clinical implant performance (CIP). The highest implant loss (29%) is found in the Transmandibular Implant group. All groups had prosthetic revisions and complications according to the CIP scale. The majority of the patients in the endosseous implant groups were subject to minor complications. The CIP-score of the Transmandibular Implant group is significantly higher than the scores of the other groups, because of the high number of lost posts. In 26;1% of the patients in this group score 4 is given, which means failure of the implant system. From this study it can be concluded that the endosseous implant systems used in this study have less surgical aftercare and a better clinical implant performance than the Transmandibular Implant system and are therefore the systems of choice for the edentulous mandible. PMID- 11856389 TI - Adhesive bonding of composite material to cast titanium with varying surface preparations. AB - The purpose of the present study was to evaluate the surface preparation effects of eight metal conditioners and an adhesive system on bonding between a prosthodontic composite material and cast titanium. Eight primers designed for conditioning base metal alloys (Acryl Bond, All-Bond 2 Primer B, Alloy Primer, Cesead II Opaque Primer, Eye Sight Opaque Primer, Metafast Bonding Liner, Metal Primer II, and MR Bond) as well as a surface modification technique (Siloc) were assessed. Disk specimens cast from titanium (T-Alloy H) were either primed with one of the eight primers or treated with the Siloc system, and then bonded with a light-activated composite material (Artglass). Bond durability was evaluated by thermocycling (4 and 60 degrees C, 1 min each, 20, 000 cycles). After thermocycling, two groups either primed with the Cesead II Opaque Primer material or treated with the Siloc system exhibited significantly greater bond strength (20.0 and 19.0 MPa) than the other groups (0.2-12.6 MPa, P < 0.05). These two systems are considered to be useful for improving bonding between the titanium and the composite material tested. PMID- 11856390 TI - Usage and status of cobalt-chromium removable partial dentures 5-6 years after placement. AB - The aim of this study was to describe the usage of cobalt-chromium (Co-Cr) removable partial dentures (RPDs) by patients 5-6 years after denture insertion and to find out the factors that affected their denture usage. A random sample of patients provided with Co-Cr RPDs from a dental teaching hospital in Hong Kong was selected. Patients were interviewed using a structured questionnaire concerning their assessment and use of the dentures. Those who had been constantly wearing their original RPDs were examined by one calibrated examiner under optimal clinical conditions. The response rate of the patients who could be contacted was 98%. The results from 189 patient interviews showed that usage of the RPDs declined with time and that half of the dentures had been discarded or replaced 5-6 years after insertion. These discarded RPDs had been in use for an average of 19.5 months. The main reason given by the patients for not using the RPDs was general dissatisfaction with the dentures in various combinations of comfort, fit and chewing ability and, less importantly, with food trapping and appearance. No statistically significant association between denture usage and their respective Kennedy classification was found. The status of the Co-Cr RPDs that had been constantly used for 5-6 years was generally good. The majority of these RPDs demonstrated fair to good cleanliness, stability and retention and had no defect of any sort. PMID- 11856391 TI - Ag, Cu, Hg and Ni ions alter the metabolism of human monocytes during extended low-dose exposures. AB - The monocyte and macrophage play an important role in the biological response to dental biomaterials. However, the effects of low-level, extended exposures of monocytes to metal ions which are known to be released from dental alloys is not known. Thus, in the current study we characterized the metabolic activity of monocytes in the presence of low doses of Ag, Cu, Hg and Ni ions for up to 4 weeks. THP-1 human monocytes were exposed in vitro to concentrations of metal ions at 1-10% of those known to be lethal during 24 h exposures. Mitochondrial function [succinic dehydrogenase (SDH) activity] and total cellular protein [bicinchoninic acid (BCA) assay] were assessed at weekly intervals during metal exposure. Each metal ion caused a unique pattern of effects from the monocytes. These effects were sometimes delayed until several weeks into the exposure (Cu, Ni). Large increases in total protein or SDH activity per cell were observed (Cu 150%, Hg 40-60%, Ni 50%), but these increases were always transient. The differences between concentrations with minimal effects and those which were lethal (8 versus 12 micromol L(-1) for Ag, 1.0 versus 1.5 micromol L(-1) for Hg) were small. Finally, concentrations which caused total suppression of cellular activity were sometimes preceded by an increased activity (Hg, Ni). We concluded that metal ions alter monocyte metabolic activity during extended exposures in vitro, but that the concentrations required are often near long-term lethal levels. Clinically, these results imply that the levels of metals released from dental alloys may be significant to monocytic function. PMID- 11856392 TI - Longevity of fixed metal ceramic bridge prostheses: a clinical follow-up study. AB - The aim of this study was to evaluate the factors that influence the longevity of fixed metal ceramic bridge prostheses, including the length of the bridge as well as the basic circumstances of the mouth. A total of 132 patients attended the clinical examination. There were 84 women (64%) and 48 men (36%). Patients had altogether 195 bridges, which included 157 short bridges (3-5 units) and 47 long bridges (6 and more units). The overall clinical examination was performed including the intra-oral radiographs. The salivary findings of the patient at baseline [flow rate of stimulated saliva, scores of mutans streptococci (SM) and lactobacilli (LB)] were registered from the patient files. In addition, the patient files were examined to receive further information concerning the complications treated in our clinic. On the basis of severe and extensive complications, the overall survival after 10 years was 84%. Long bridges had lower survival than the shorter ones (P=0.04). A low secretion of saliva recorded before the prosthetic treatment decreased the survival of the bridges (P=0.12) and the survival of the bridges was likewise reduced by high scores of LB and SM (P=0.07). Age of the patient did not influence the survival. It was shown here that long bridges (5 and more units) have a lower survival than the short bridges. It can be concluded, however, that the low secretion of saliva and the high scores of lactobacilli and SM have a remarkable influence on complications and these facts should therefore be taken into careful consideration in choosing the patients for the prosthetic treatment with fixed prosthodontics. PMID- 11856393 TI - Properties of athletic mouth protectors and materials. AB - The properties of a variety of mouth protectors and sheet materials used to fabricate custom mouth protectors were determined in order to recommend limits for a specification. Hardness, water sorption, water solubility, impact absorption, and tear strength were measured, and limits for these properties were suggested. PMID- 11856394 TI - Forces resisting jaw displacement in relaxed humans: a predominantly viscous phenomenon. AB - Forces opening the relaxed human jaw are resisted by intrinsic restraints, including passive tensions in the jaw-closing muscles. These muscle tensions have been modelled as viscoelastic elements, and static measurements suggest their elastic portions contribute approximately a total of 5 N resistance at wide gape. As the viscous damping properties of muscles which affect the jaw's dynamic behaviour are unknown, we measured the jaw opening force required to reach maximum gape during fast and slow opening in six relaxed subjects. These data were then incorporated in a dynamic mathematical jaw model to determine the damping properties of the masticatory system. During the 3 and 8 s opening trials, forces increased with gape (6.7 +/- 3.3 and 3.9 +/- 2.3 N, respectively, at 50% gape) and reached their maxima at wide gape (19.9 +/- 4.5 and 13.2 +/- 4.4 N, respectively). The muscle damping constant needed by the model to emulate these results was 150 Nsm(-1), approximately 25% lower than the calculated critical damping constant. This study suggests low forces are required to open the jaw in relaxed humans, and that jaw viscosity, not elasticity, provides the major resistance to motion. PMID- 11856395 TI - Acoustic characteristics of sounds from temporomandibular joints with and without effusion: an MRI study. AB - Joint effusion has been associated with temporomandibular joint (TMJ) pain but can only be diagnosed by magnetic resonance imaging (MRI). For screening of patients with suspected effusion a simple and less expensive method would be desirable. We recorded joint sounds during jaw opening and closing movement from 34 TMJs with internal derangement (ID). Seventeen joints had joint effusion seen on MRI. Spectrograms of the sounds were displayed as waterfall plots showing profiles of the consecutive Hamming windows. If the profiles were similar, as judged by initial evaluation, the displayed pattern was classified as stable. If some profiles were distinctly deviating in their pattern, this was classified as unstable. Joints with effusion showed unstable sound pattern more often than joints without effusion (P < 0.001). It was concluded that TMJ sound analyses have a potential to identify joints with effusion based on their unstable sound pattern. PMID- 11856396 TI - Dentine hypersensitivity: the effects of brushing toothpaste on etched and unetched dentine in vitro. AB - Although many people have exposed dentine, only a percentage exhibit symptoms of dentine hypersensitivity. This has been ascribed to opening and closing of the dentinal tubules by for example, smear layer changes or tubular occlusion. The aims of this study were to examine the surface morphological changes of etched and unetched dentine in vitro, attributed to the effects of toothbrushing with and without toothpastes designed for the alleviation of dentine hypersensitivity. A total of 96 etched and 96 unetched human dentine specimens were brushed with various toothpastes and water for 1, 2, 5 or 10-min periods in a toothbrushing machine and subsequently examined under scanning electron microscopy for surface changes. Analyses of brushed etched specimens demonstrated that time and treatment were significant variables (P < 0.05) for tubule occlusion. Further, the interaction between time and treatment was significant (P < 0.05). The artificial silica based paste was significantly better for all time intervals at occluding the dentine tubules. All toothpastes investigated caused dynamic changes to the smear layer of the unetched dentine, opening tubules. However, the artificial silica based paste resulted in occluded rather than patent tubules. All of the pastes evaluated had the capacity to remove the smear layer but some could then occlude tubules through the contained abrasives. PMID- 11856398 TI - The influence of selective grinding on the thickness discrimination threshold of patients wearing complete dentures. AB - The influence of selective grinding on the ability of patients wearing complete dentures to discriminate thicknesses occlusally was evaluated. The study included two groups of white Caucasian patients, matched for age, sex and duration of edentulism; 12 wearing traditional complete dentures and 12 wearing maxillary complete denture and mandibular implant-retained overdenture (MIR-OVD). The ability to discriminate thickness was evaluated before and after selective grinding. In both groups, sensitivity improved after selective grinding. Subjects wearing MIR-OVD showed a better ability to discriminate thickness than those rehabilitated with traditional complete dentures. The thickness discrimination threshold might be a suitable method to evaluate complete dentures fit. PMID- 11856397 TI - Fracture characteristics of carbon fibre, ceramic and non-palladium endodontic post systems at monotonously increasing loads. AB - A carbon fibre post system, three non-palladium and one palladium metal post systems, two ceramic post systems, and a metal post system with a ceramic core were studied in vitro. The control group consisted of root-filled test teeth without posts. The test teeth were identical artificial roots of an upper central incisor made from a posterior composite whose module of elasticity was similar to that of natural dentine. All posts were cemented in the roots using Panavia 21 TC. Subsequently, standardized full crowns were cemented onto all roots. On a universal testing machine, the test teeth were loaded palatally at monotonously increasing loads until root fracture. The highest mean fracture loads were found for the carbon fibre post system (312.5 +/- 58.8 N). The fracture load of non palladium metal posts (242.3-300.4 N) did not differ significantly from that of the Perma-dor post (265.9 N), which does contain palladium. Values of 300.3 +/- 89.3 N (aluminium oxide ceramics) and 193.5 +/-57.0 N (zirconia ceramics) were found for the ceramic posts. The control group exhibited a fracture load of 228.8 +/- 35.7 N. The mean distance between the vestibular end of the fracture gap and the point of force application was between 10.1 +/- 2.3 and 14.7 +/- 1.2 mm. PMID- 11856399 TI - The effect of variation in occlusal loading on the development of abfraction lesions: a finite element study. AB - Abfraction or non-carious cervical tooth loss is poorly understood and one factor thought to contribute to the development of these lesions is the effect of occlusal loading. The aim of this study was to examine the effect that varying the position of an occlusal load would have on the stress contour in the cervical region of a lower second premolar using a two-dimensional plane strain finite element model. A 500 N load was applied vertically to either of the cusp tips or in various positions along the cuspal inclines. It was found that loads applied to the inner aspects of the buccal or the lingual cuspal inclines produced maximum principal stress values of up to 358 MPa that exceed the known failure stresses for enamel. PMID- 11856400 TI - Occlusal contact area (OCA) wear of two new composite restoratives. AB - The occlusal contact area wear resistance of two new composite restoratives (Z250 and P60) was investigated and compared with two existing materials (Silux and Z100) using a reciprocal compression-sliding test apparatus. Six wear specimens were made for each material and subjected to wear testing at 20 MPa with artificial saliva as lubricant. Wear depth (microm) was measured using profilometry every 20, 000 cycles up to 120 000 cycles. Results were analysed using ANOVA/Scheffe's test (P < 0.05) and Pearson's correlation. The effects of cyclic loading on wear and counter-body loss were material dependent. Silux had the lowest wear at all cyclic intervals. At 20 000 and 40 000 cycles, Silux had significantly less wear than all the other composites evaluated, and Z100 had significantly less wear than Z250 and P60. After 60 000 cycles, significant differences in wear was observed only between Silux and the other composites. Correlation between number of cycles and wear/counter-body loss, and between wear and counter-body were significant and positive. The microfilled composite Silux had significantly better OCA wear resistance than the minifilled composites Z100, Z250 and P60. PMID- 11856401 TI - In vivo mandibular elastic deformation during clenching on pivots. AB - Lower rigidly connected long span bridges supported by natural abutments or implants sometimes become loose, come off, or fracture after a period of usage. Many reasons have been discussed for these failures. However, few researchers have shown the influence of mandibular elastic deformation on the abutments, although this influence is likely to produce a distortion force between the abutment and prosthesis. Therefore, this study measured the elastic deformation of the human mandibular arch during clenching on pivots by using charge-coupled device (CCD) cameras and an image analysing system. When the subjects clenched on the canines (unilaterally or bilaterally) or bilateral second molars, no mandibular deformation was found; whereas when the subjects clenched on the unilateral second molars, the mandibular arch on the non-pivot side moved upward and inward and the straight line distances between the right and left measurement points decreased by 0.2 mm. The magnitude of deformation is smaller than the depressible limit of periodontal membrane. This suggests that the influence of mandibular deformation on the connected prosthesis is negligible in the case of the natural root supported long span bridge but should probably be considered in the case of the implant supported bridge. PMID- 11856403 TI - A brief analysis of clinical pharmacy interventions undertaken in an Australian teaching hospital. AB - Selected clinical pharmacy interventions undertaken during a 30-day data capture period were analysed, seeking to gain a greater understanding of the nature of the drug-related problems involved. Pharmacists were asked to record only interventions that were of potentially major significance. A total of 67 interventions were submitted for analysis. In 28 cases (41.7% of the initial total) the intervention reports were excluded from further analysis after initial review. For the remaining 39 interventions, 20 patients (51%) were under the care of a medical unit, and cardiovascular/antithrombotic agents accounted for 17 reports (43.5%). The majority of interventions were implemented at the time of inpatient medication order review by the clinical pharmacist (n=25, 64%). The most common category of drug-related problem addressed in the interventions related to the prescription of inappropriately high doses of the correct drug for the patient (n=17, 43.6%). Deficiencies in technical knowledge accounted for less than 25% of all cases. PMID- 11856404 TI - General practitioner-hospital communications: a review of discharge summaries. AB - The objective of this study was to examine the reliability, effectiveness, accuracy and timeliness of hospital to general practitioner (GP) information transfer by discharge summaries produced in a general public hospital in New South Wales, Australia. A retrospective audit of 569 patient discharge summaries and related medical records with a targeted GP interview was performed to determine receipt and clinical value of the recorded information. The main outcome measure was the number and quality of discharge summaries received by patient-nominated GPs. Summaries written for patients discharged from hospital were estimated to be received by the patient-nominated GP in 27.1% of cases. Discharge summaries audited were rated as being 63.6% accurate, with errors occurring in all facets of production. The current method of discharge summary production and distribution is unacceptable. The high number of errors (36.4%) and the low rate of receipt (27.1%), indicates that resources invested in the production of the discharge summary could be better utilized to improve information transfer. PMID- 11856405 TI - A 30-month study of patient complaints at a major Australian hospital. AB - Health practitioners often regard complaints about the quality of patient care in a negative light. However, complaints can indicate strategies to improve care. Therefore, an audit was undertaken of all formal complaints about patient care at a major Australian hospital over a 30-month period. The profile of complainants, the reasons for complaints, and the outcome were analysed. A total of 1308 complaints, concerning the care of 1267 patients, were received. The complaint rate was 1.12 per 1000 occasions of service. In all, 57% of complaints were lodged by advocates and 71% of complaints related to poor communication or to the treatment provided. In 97% of occasions, an explanation and/or an apology resulted. To date, no complaint has proceeded to litigation. Complaints are potentially useful quality assurance tools and can identify remediable system flaws. Health professionals and employers should understand why patients complain and be able to respond appropriately. PMID- 11856407 TI - Quality assessment in private practice: the clinician as service provider. AB - Assessment of quality in the private practice setting may be difficult. The author has taken the view that service delivery is a key outcome of private clinician practice. A method of assessing service delivery in a private consultant physician practice setting is described. PMID- 11856406 TI - The application of statistical process control charts to the detection and monitoring of hospital-acquired infections. AB - The monitoring of infection control indicators including hospital-acquired infections is an established part of quality maintenance programmes in many health-care facilities. However, surveillance data use can be frustrated by the infrequent nature of many infections. Traditional methods of analysis often provide delayed identification of increasing infection occurrence, placing patients at preventable risk. The application of Shewhart, Cumulative Sum (CUSUM) and Exponentially Weighted Moving Average (EWMA) statistical process control charts to the monitoring of indicator infections allows continuous real-time assessment. The Shewhart chart will detect large changes, while CUSUM and EWMA methods are more suited to recognition of small to moderate sustained change. When used together, Shewhart and EWMA methods are ideal for monitoring bacteraemia and multiresistant organism rates. Shewhart and CUSUM charts are suitable for surgical infection surveillance. PMID- 11856408 TI - Recovery after stroke: a qualitative perspective. AB - The purpose of this study was to determine the impact of stroke on survivors of the condition and to identify their physical and psychosocial needs in rural and regional settings. Data were collected via focus group interviews with stroke survivors, carers and key informants. Data were managed using NUD*IST and analysed using a content analysis method identifying major themes related to the impact of living in the community after having a stroke. It was found that stroke survivors suffered severe physical and emotional effects. The findings also identified the vulnerability of this group and a lack of organised, on-going psychosocial and rehabilitative support. Recommendations are made to enhance the current management of stroke after the acute and subacute phases. PMID- 11856409 TI - An evidence-based medicine fellowship in a children's teaching hospital. AB - This paper reports the introduction of an evidence-based medicine fellowship in a children's teaching hospital. The results are presented of a self-reported 'evidence-based medicine' questionnaire, the clinical questions requested through the information retrieval service are outlined and the results of an information retrieval service user questionnaire are reported. It was confirmed that clinicians have frequent clinical questions that mostly remain unanswered. The responses to four questions with 'good quality' evidence-based answers were reviewed and suggest that at least one-quarter of doctors were not aware of the current best available evidence. There was a high level of satisfaction with the information retrieval service; 19% of users indicated that the information changed their clinical practice and 73% indicated that the information confirmed their clinical practice. The introduction of an evidence-based medicine fellowship is one method of disseminating the practice of evidence-based medicine in a tertiary children's hospital. PMID- 11856410 TI - A review of clinical risk management. AB - Current literature has a plethora of information regarding medical errors, what causes them, how to reduce them and the impact they have on patients, staff and health-care organisations. The objective of this paper was to give a summary of the current findings of recent studies and articles, including worldwide initiatives to reduce medical errors, the trend to identify system issues that result in medical error rather than the focus on individual blame, the impact of technological changes and current medical advances, litigation and law suits. PMID- 11856411 TI - Quality of health services and early postpartum discharge: results from a sample of non-English-speaking women. AB - Few studies have explored the knowledge, perceptions and satisfaction of ethnic women in relation to early obstetric discharge. The aim of this study was to form a profile of Vietnamese mothers who opt for early postpartum discharge, and to identify factors associated with those decisions. Focus group discussions, in depth interviews and survey questionnaires were utilized to collect information from a sample of 160 mothers. Sixty mothers (38%) opted for early discharge (< 48 h). Factors strongly associated with early discharge were poor comprehension of English (OR=6.58, 95% CI=1.81, 24.02), very low level of education (OR=3.10, 95% CI=1.55, 6.20) and first time mother (OR=2.70, 95% CI=1.15, 4.27). The in-depth interviews identified further factors driving early discharge, including fear and anxiety when having to approach staff for assistance, and perceptions of disempowerment within culturally unfamiliar hospital surroundings. The findings in this study ought to alert administrators and clinicians to possible negative drivers for the choice of early postpartum discharge by Vietnamese mothers. PMID- 11856412 TI - Postnatal depression: use of health services and satisfaction with health-care providers. AB - The objective of this study was to compare health-care use and satisfaction with health-care providers between depressed and non-depressed women in the first 4 months after childbirth. Sixteen weeks after delivery a questionnaire, which included the Edinburgh Postnatal Depression Scale (EPDS) and items about health care use and satisfaction, was mailed to women who attended the antenatal clinic, Royal Women's Hospital, Brisbane. Completed questionnaires were returned by 574 (86.4%) of the 664 women surveyed. During the study period most women (91%) visited a general practitioner at least once and 117 (12%) saw their doctor on five or more occasions. A total of 118 (20.7%) scored above 12 on the EPDS. Depressed women were more likely to visit a psychiatrist (OR, 9.2; 95% CI, 4.3 19.6), social worker (OR, 6.1; 95% CI, 3.3-11.1), postnatal depression group (OR, 4.0; 95% CI, 1.3-12.6), paediatrician (OR, 2.5; 95% CI, 1.6-3.9), or a general practitioner (OR, 2.1; 95% CI, 1.4-3.2) than non-depressed women. Twenty-two (18.5%) of the depressed women had contact with a psychiatrist. Compared with non depressed women, those scoring above 12 on the EPDS were less satisfied with the services of general practitioners (P=< 0.000), paediatricians (P=0.002), Nursing Mothers' Associations of Australia (P=0.043) and obstetricians (P=0.045). Postpartum depression leads to an increase use of health-care services and has a negative effect on satisfaction with some services. PMID- 11856413 TI - Prevention of orthopaedic wound infections: a quality improvement project. AB - Using clinical practice improvement methodology, a project was undertaken to reduce the incidence of surgical wound infections following elective hip and knee replacement surgery. A team was established, key measures for improvement were identified, strategies for change were developed and an action plan was implemented. Outcomes for this project included a reduction in the rate of clean surgical wound infection for joint replacement surgery from 28% to zero. Average length of stay for total hip replacement surgery was reduced from 13.9 to 9.3 days and from 14.6 to 10.4 days for total knee replacement surgery. Guidelines for patient selection were developed along with a protocol for the management of preparation to prevent urinary tract infections. Post-discharge surveillance and a preoperative rehabilitation and exercise programme have been implemented. There is potential for wider uptake and implementation of the quality principles described herein. PMID- 11856415 TI - Facilitating best practice: transferring the lessons of the Clinical Support Systems Program. AB - The Clinical Support Systems Program (CSSP) includes the management of clinical practice using clinical and consumer pathways, outcome and performance indicators, clinical measurement and review in a continuous improvement cycle using the best available extant evidence. The Royal Australasian College of Physicians is testing the CSSP model through four consortia around Australia. There are 17 project sites in three States. The funded projects address major clinical problems including congestive heart failure and acute coronary syndromes, acute stroke management, and colorectal cancer care. There is some early evidence of the CSSP influencing change in areas beyond the bounds of the project settings and the College has developed a plan to promote wider adoption of best practice. This approach recognises the College's role in providing Fellows with the practical tools of quality improvement, the means to collect data and compare their practice to other clinicians, while traversing the appropriate educational framework. PMID- 11856414 TI - Are routine chest X-rays for students entering university worthwhile? AB - Mass radiographic screening for tuberculosis has lost favour in many countries. The aim of this study was to determine whether the continued practice of such screening of prospective students at the University of the West Indies was warranted by assessing the yield and the cost of the programme in our setting. In a cross- sectional retrospective study, 12,662 chest X-ray reports collected over the period 1989-1997 were studied. No active case of tuberculosis was detected. Three students reported a previous history of tuberculosis and 10 students had a positive family history of tuberculosis. Three hundred and ninety-nine clinically insignificant abnormalities were reported, such as mild scoliosis and calcified foci. Routine radiological screening of prospective students at the University of the West Indies for tuberculosis has an extremely low yield, places the students at unnecessary risk of radiation exposure and should be discontinued. PMID- 11856416 TI - Prioritising quality. AB - The deluge of new initiatives, proposals and recommendations for solving the problems required to improve patient safety and quality of health care continues unabated. Implementing the proposed solutions for improving patient safety and quality of care requires setting priorities for action throughout the different levels of the health-care system. Currently, we face the dilemma of having to set priorities from the plentiful themes that are intuitively sensible and well accepted. There is an increasing number of examples of using systematic processes which contain explicit and transparent criteria for priority setting in patient safety and quality initiatives. These should be more widely adopted and become a key requirement for all future proposals for improving patient safety and quality of health care. PMID- 11856418 TI - The estimation of population differentiation with microsatellite markers. AB - Microsatellite markers are routinely used to investigate the genetic structuring of natural populations. The knowledge of how genetic variation is partitioned among populations may have important implications not only in evolutionary biology and ecology, but also in conservation biology. Hence, reliable estimates of population differentiation are crucial to understand the connectivity among populations and represent important tools to develop conservation strategies. The estimation of differentiation is c from Wright's FST and/or Slatkin's RST, an FST -analogue assuming a stepwise mutation model. Both these statistics have their drawbacks. Furthermore, there is no clear consensus over their relative accuracy. In this review, we first discuss the consequences of different temporal and spatial sampling strategies on differentiation estimation. Then, we move to statistical problems directly associated with the estimation of population structuring itself, with particular emphasis on the effects of high mutation rates and mutation patterns of microsatellite loci. Finally, we discuss the biological interpretation of population structuring estimates. PMID- 11856419 TI - Prevailing triple infection with Wolbachia in Callosobruchus chinensis (Coleoptera: Bruchidae). AB - Prevailing triple infection with three distinct Wolbachia strains was identified in Japanese populations of the adzuki bean beetle, Callosobruchus chinensis. When a polymerase chain reaction (PCR) assay was conducted using universal primers for ftsZ and wsp, Wolbachia was detected in all the individuals examined, 288 males and 334 females from nine Japanese populations. PCR-restriction fragment length polymorphism (RFLP) analysis of cloned wsp gene fragments from single insects revealed that three types of wsp sequences coexist in the insects. Molecular phylogenetic analysis of the wsp sequences unequivocally demonstrated that C. chinensis harbours three phylogenetically distinct Wolbachia, tentatively designated as wBruCon, wBruOri and wBruAus, respectively. Diagnostic PCR analysis using specific primers demonstrated that, of 175 males and 235 females from nine local populations, infection frequencies with wBruCon, wBruOri and wBruAus were 100%, 96.3% and 97.0%, respectively. As for the infection status of individuals, triple infection (93.7%) dominated over double infection (6.1%) and single infection (0.2%). The amounts of wBruCon, wBruOri and wBruAus in field-collected adult insects were analysed by using a quantitative PCR technique in terms of wsp gene copies per individual insect. Irrespective of original populations, wBruCon and wBruOri (107 -108 wsp copies/insect) were consistently greater in amount than wBruAus (106 -107 wsp copies/insect), suggesting that the population sizes of the three Wolbachia strains are controlled, although the mechanism is unknown. Mating experiments suggested that the three Wolbachia cause cytoplasmic incompatibility at different levels of intensity. PMID- 11856420 TI - Genetic variation in original and colonizing Drosophila buzzatii populations analysed by microsatellite loci isolated with a new PCR screening method. AB - A new polymerase chain reaction-based screening method for microsatellites is presented. Using this method, we isolated 12 microsatellite loci from Drosophila buzzatii, two of which were X-linked. We applied the other 10 microsatellite loci to the analysis of genetic variation in five natural populations of D. buzzatii. Two populations were from the species' original distribution in Argentina, whereas the other three were from Europe (two) and Australia that were colonized 200 and 65 years ago, respectively. Allelic variation was much larger in the original populations than in the colonizing ones and there was a tendency to decreased heterozygosity in the colonizing populations. We used three different statistical procedures for detecting population bottlenecks. All procedures suggested that the low variability in the populations in the Old World was not the result of the recent population decline, but was due to a founder effect followed by a population expansion. In fact, one procedure which detects population expansions and declines based on the genealogical history of microsatellite data suggested that an expansion had taken place in all the colonized populations. PMID- 11856421 TI - Extensive exchange of fungal cultivars between sympatric species of fungus growing ants. AB - Fungal cultivars of fungus-growing ants (Attini, Formicidae) are carried by dispersing queens from parent to offspring nest. This vertical cultivar transmission between generations is thought to result in long-term ant-fungus coevolution and selection for beneficial cultivar traits that maximize harvests and thus colony productivity. In contrast to this traditional view of vertical cultivar transmission, frequent horizontal cultivar transmission between ant species is implicated by a phylogenetic analysis of 72 cultivars propagated by two fungus-growing ant species coexisting sympatrically in central Panama. The two ant species are specialized on the same group of closely related cultivars, but in six of 12 cultivar clades identifiable within this group, cultivars from both ant species were united in the same clade. Five of these 'mixed' clades were supported by bootstrap values of about 90% or higher. In one instance, colonies from the two ant species cultivated the same, genetically identical, cultivar clone. These phylogenetic patterns indicate that: (i) cultivar exchanges between the two ant species occur routinely throughout ecological time; and that (ii) coevolutionary processes between ants and their fungi are more diffuse than previously assumed. Because the two ant species are specialized on a narrow group of closely related cultivars that they regularly exchange among each other, but not with other sympatric ant species, cultivar exchanges are constrained, most likely, by ant preferences for their own cultivar group or by stringent selection against transitions of ant lineages to distantly related cultivars. PMID- 11856422 TI - Temporal change in genetic structure and effective population size in steelhead trout (Oncorhynchus mykiss). AB - There is a wealth of published molecular population genetic studies, however, most do not include historic samples and thus implicitly assume temporal genetic stability. We tested for changes in genetic diversity and structure in three populations of steelhead trout (Oncorhynchus mykiss) from a northern British Columbia watershed using seven microsatellite loci over 40 years. We found little change in genetic diversity (mean allele numbers and observed and expected heterozygosity), despite large variation in the estimated numbers of steelhead returning to the watershed over the same time period. However, the temporal stability in genetic diversity is not reflected in population structure, which appears to be high among populations, yet significantly variable over time. The neighbour-joining tree showed that, overall, two of the populations (Zymoetz and Kispiox) clustered separately from the third (Babine); a finding which was not consistent with their geographical separation. The clustering pattern was also not temporally consistent. We used the temporal method to estimate the effective number of breeders (Nb ) for the three populations; our values (Nb = 17-102) were low for the large and presumed vigorous populations of steelhead trout sampled. The low Nb values were also not consistent with the generally high genetic diversity estimates, suggesting the possibility of intermittent gene flow among the three populations. The use of temporal analyses in population genetic samples should be a priority; first, to verify observed patterns in contemporary data, and second, to build a dataset of temporal analyses to allow generalizations to be made concerning temporal genetic stability and effective population size in natural populations. PMID- 11856423 TI - DNA from bird-dispersed seed and wind-disseminated pollen provides insights into postglacial colonization and population genetic structure of whitebark pine (Pinus albicaulis). AB - Uniparentally inherited mitochondrial (mt)DNA and chloroplast (cp)DNA microsatellites (cpSSRs) were used to examine population genetic structure and biogeographic patterns of bird-dispersed seed and wind-disseminated pollen of whitebark pine (Pinus albicaulis Engelm.). Sampling was conducted from 41 populations throughout the range of the species. Analyses provide evidence for an ancestral haplotype and two derived mtDNA haplotypes with distinct regional distributions. An abrupt contact zone between mtDNA haplotypes in the Cascade Range suggests postglacial biogeographic movements. Among three cpSSR loci, 42 haplotypes were detected within 28 cpSSR sample populations that were aggregated into six regions. Analysis of molecular variance (amova) was used to determine the hierarchical genetic structure of cpSSRs. amova and population pairwise comparisons (FST ) of cpSSR, and geographical distribution of mtDNA haplotypes provide insights into historical changes in biogeography. The genetic data suggest that whitebark pine has been intimately tied to climatic change and associated glaciation, which has led to range movements facilitated by seed dispersal by Clark's nutcracker (Nucifraga columbiana Wilson). The two hypotheses proposed to explain the genetic structure are: (i) a northward expansion into Canada and the northern Cascades in the early Holocene; and (ii) historical gene flow between Idaho and the Oregon Cascades when more continuous habitat existed in Central Oregon during the late Pleistocene. Genetic structure and insights gained from historical seed movements provide a basis on which to develop recovery plans for a species that is at risk from multiple threats. PMID- 11856424 TI - Phylogeographical structure and regional history of the dusky-footed woodrat, Neotoma fuscipes. AB - The dusky-footed woodrat, Neotoma fuscipes, is a medium-sized rodent that inhabits low elevation woodland habitats along the Pacific coast of North America from Oregon, throughout California and into Baja California. Analyses of mitochondrial sequence variation throughout the distribution reveal substantial phylogeographical structure within N. fuscipes. The major mitochondrial lineages are largely concordant with previously identified morphological subdivisions within the taxon. The geographical distribution of distinct clades suggests that a combination of topographic barriers and the expansion and contraction of suitable habitat during the past 2 million years, especially along particular mountain ranges, have played a major role in the diversification of N. fuscipes. Furthermore, relatively low levels of genetic variation across the northern half of the distribution suggest that dusky-footed woodrats may have only recently expanded into this region. PMID- 11856425 TI - Adult habitat preferences, larval dispersal, and the comparative phylogeography of three Atlantic surgeonfishes (Teleostei: Acanthuridae). AB - Although many reef fishes of the tropical Atlantic are widely distributed, there are large discontinuities that may strongly influence phylogeographical patterns. The freshwater outflow of the Amazon basin is recognized as a major barrier that produces a break between Brazilian and Caribbean faunas. The vast oceanic distances between Brazil and the mid-Atlantic ridge islands represent another formidable barrier. To assess the relative importance of these barriers, we compared a fragment of the mitochondrial DNA (mtDNA) cytochrome b gene among populations of three species of Atlantic surgeonfishes: Acanthurus bahianus, A. chirurgus and A. coeruleus. These species have similar life histories but different adult habitat preferences. The mtDNA data show no population structure between Brazil and the mid-Atlantic islands, indicating that this oceanic barrier is readily traversed by the pelagic larval stage of all three surgeonfishes, which spend approximately 45-70 days in the pelagic environment. The Amazon is a strong barrier to dispersal of A. bahianus (d = 0.024, phiST = 0.724), a modest barrier for A. coeruleus (phiST = 0.356), and has no discernible effect as a barrier for A. chirurgus. The later species has been collected on soft bottoms with sponge habitats under the Amazon outflow, indicating that relaxed adult habitat requirements enable it to readily cross that barrier. A limited ability to use soft bottom habitats may also explain the low (but significant) population structure in A. coeruleus. In contrast, A. bahianus has not been collected over deep sponge bottoms, and rarely settles outside shallow reefs. Overall, adult habitat preferences seem to be the factor that differentiates phylogeographical patterns in these reef-associated species. PMID- 11856426 TI - Detection of genetically divergent clone mates in apomictic dandelions. AB - This study aims to identify genetically diverged clone mates in apomictic dandelions. Clone mates are defined as individuals that may have diverged as a result of mutation accumulation and that have undergone only clonal reproduction since their most recent common ancestor. Based on distinctive morphology and an aberrant and rare chloroplast haplotype, northwest European individuals of Taraxacum section Naevosa are well suited for the detection of clonal lineages in which mutation has occurred. In the case of strictly clonal reproduction, nuclear genetic variability was expected to be hierarchically organized. Nucleotide polymorphisms in internal transcribed spacer (ITS) sequences, however, were incompatible with a clonal structure of the Norwegian individuals, probably due to persistent ancestral polymorphisms that pre-date the origin of the Naevosa clone. This interpretation is supported by the presence of ITS variants in section Naevosa that were also found in distantly related dandelions. In contrast to the ITS sequence data, amplified fragment length polymorphisms (AFLPs), isozymes and microsatellites strongly supported the contention of prolonged clonal reproduction and mutation accumulation in Norwegian Naevosa. Because these markers are generally considered to be more variable and more rapidly evolving than ITS sequences, mutations in these markers probably evolved after the origin of the clone. Within the Norwegian clone, a surprising number of markers distinguished the clone mates. As a consequence, incorporation of mutation in the detection of clone mates is anticipated to have a big impact on estimates of size, geographical range and age of clones as well as on experimental designs of studies of clonal plants. PMID- 11856427 TI - Phylogeography of the African buffalo based on mitochondrial and Y-chromosomal loci: Pleistocene origin and population expansion of the Cape buffalo subspecies. AB - Population genetics and phylogeography of the African buffalo (Syncerus caffer) are inferred from genetic diversity at mitochondrial D-loop hypervariable region I sequences and a Y-chromosomal microsatellite. Three buffalo subspecies from different parts of Africa are included. Nucleotide diversity of the subspecies Cape buffalo at hypervariable region I is high, with little differentiation between populations. A mutation rate of 13-18% substitutions/million years is estimated for hypervariable region I. The nucleotide diversity indicates an estimated female effective population size of 17 000-32 000 individuals. Both mitochondrial and Y-chromosomal diversity are considerably higher in buffalo from central and southwestern Africa than in Cape buffalo, for which several explanations are hypothesized. There are several indications that there was a late middle to late Pleistocene population expansion in Cape buffalo. This also seems to be the period in which Cape buffalo evolved as a separate subspecies, according to the net sequence divergence with the other subspecies. These two observations are in agreement with the hypothesis of a rapid evolution of Cape buffalo based on fossil data. Additionally, there appears to have been a population expansion from eastern to southern Africa, which may be related to vegetation changes. However, as alternative explanations are also possible, further analyses with autosomal loci are needed. PMID- 11856428 TI - Multiple paternity in loggerhead turtle (Caretta caretta) nests on Melbourne Beach, Florida: a microsatellite analysis. AB - Many aspects of sea turtle biology are difficult to measure in these enigmatic migratory species, and this lack of knowledge continues to hamper conservation efforts. The first study of paternity in a sea turtle species used allozyme analysis to suggest multiple paternity in loggerhead turtle (Caretta caretta) clutches in Australia. Subsequent studies indicated that the frequency of multiple paternity varies from species to species and perhaps location to location. This study examined fine-scale population structure and paternal contribution to loggerhead clutches on Melbourne Beach, FL, USA using microsatellite markers. Mothers and offspring from 70 nests collected at two locations were analysed using two to four polymorphic microsatellite loci. Fine scale population differentiation was not evident between the sampled locations, separated by 8 km. Multiple paternity was common in loggerhead nests on Melbourne Beach; 22 of 70 clutches had more than one father, and six had more than two fathers. This is the first time that more than two fathers have been detected for offspring in individual sea turtle nests. Paternal genotypes could not be assigned with confidence in clutches with more than two fathers, leaving the question of male philopatry unanswered. Given the high incidence of multiple paternity, we conclude that males are not a limiting resource for this central Florida nesting aggregate. PMID- 11856429 TI - Itraconazole in the treatment of tinea capitis in children. Case reports with long-term follow-up evaluation. Review of the literature. AB - Although griseofulvin is considered the standard treatment of tinea capitis in children, alternatives are being investigated. Our purpose was to determine the efficacy of itraconazole for kerion and noninflammatory tinea capitis. An open label study was performed on five patients. It was planned to treat them with itraconazole until they were mycologically and clinically cured. A 28-112-day course of 100 mg itraconazole daily, combined with a topical antifungal treatment resulted in clinical and mycological cure in all children. One child stopped taking itraconazole after 28 days, before it was clinically cured, because of nausea. Nevertheless, this child also achieved clinical and mycological cure. No other side-effects were reported. In long-term follow-up evaluation of between 2 and 3.5 years no recurrence or reinfection was observed. There was complete regrowth of hair, even after kerion. These findings and the review of the literature suggest that itraconazole offers an alternative to griseofulvin for the treatment of tinea capitis in children, although it is more expensive and not approved by German state authorities for this indication. PMID- 11856430 TI - First isolation of Trichophyton raubitschekii (syn. T. rubrum) in Europe. AB - Trichophyton raubitschekii is a rare dermatophyte which belongs to the Trichophyton rubrum species complex. Since 1981, only a few cases of dermatophytosis due to this anthropophilic causative agent were published. In this paper the authors report the first cases of Tinea corporis caused by Trichophyton raubitschekii (syn. T. rubrum) in Europe. The patients, one immigrant from Ghana and three from Cameroon, had typical lesions of tinea corporis. Four strains were isolated and characterized by conventional and molecular methods. On morphological and physiological grounds the isolates were identified as T. raubitschekii by the following phenotypical features: (1) velvety colony texture; (2) brown pigment; (3) abundant macroconidia and (4) positive urease activity. Molecular diagnostics were performed by single strand conformation polymorphism (SSCP) and sequence analysis of the ATPase9 intron of the mitochondrial (mt) DNA and the internal transcribed spacer (ITS) region of the rDNA, respectively. The ITS sequences and SSCP patterns of the ATPase9 intron were found to be identical among the four strains and also when compared to reference strains of T. rubrum. As shown in the present paper, T. raubitschekii is genetically identical to T. rubrum but differs in some phenotypical characteristics. Since misidentification with other dark-coloured dermatophyte variants is possible, medical mycologists should bear in mind the special morphological characteristics of T. raubitschekii (syn. T. rubrum) for future identifications. PMID- 11856431 TI - Recovery of Candida dubliniensis from sputum of cystic fibrosis patients. AB - Yeasts have been cultivated in a high percentage of sputum samples from patients with cystic fibrosis (CF). In cases of Candida dubliniensis a high recovery rate (15-25%) has so far only been reported from the oral cavity of HIV-infected persons. When studying sputum samples of patients suffering from cystic fibrosis (n = 54) we could repeatedly observe C. dubliniensis in 6 patients (11.1%) by using a complex isolation procedure involving the use of Staib agar. This revealed that the prevalence rate was significantly higher than the estimated overall prevalence for all non-HIV infected patients (0.8%) currently reported. PMID- 11856432 TI - Antifungal activity of Crotalus durissus cumanensis venom. AB - The susceptibility to Crotalus venom of 14 yeast and 10 mould fungal isolates was assessed. This venom was tested in a standardized well diffusion test, using 400 microg/20 microl well. The percentage of susceptibility to yeast isolates was 78.6% (> 8 mm); that for filamentous isolates was 50% (> 8 mm). PMID- 11856433 TI - Diagnosis of systemic Candida infections in patients of the intensive care unit. Significance of serum antigens and antibodies. AB - The combined detection of Candida antigen and antibody for the determination of systemic Candida infections (SCI) was investigated. One hundred and four patients from the intensive care unit (ICU) were analysed. Seventeen of the patients were suspected of having SCI, based on clinical and laboratory criteria. In these patients, Candida antigens and antibodies were analysed extensively. Ten patients had a positive Candida antigen (titre >1:16) determined by the latex agglutination assay Cand-Tec(R) and their median antibody titre was 1:160 in the indirect haemagglutination test (HAT). Seven antigen-negative patients had a median titre of 1:1280 (HAT). Forty-one of 42 colonized control patients had negative antigen titres and a median antibody titre of 1:160. The sensitivities and specificities were 58.8% and 97.6% for antigenemia, and 52.9% and 85.7% for antibody detection. These values reached 100.0% and 83.3%, respectively, when the results of both tests were combined. This indicates a high degree of concordance between serological results (Candida antigen and/or antibodies) and clinical presentation. We conclude, that the combined investigation of antigen and antibody titres might be a helpful tool in the characterization of SCI in ICU patients, if antigen titres are >or=1:16 or antibody titres (HAT) are >or=1:640. PMID- 11856434 TI - Onychomycosis in Northern Greece during 1994-1998. AB - In the 5-year period 1994-1998, 13957 patients were examined in the Mycological Laboratory of the State Hospital for Skin and Venereal Diseases, in Thessaloniki, Greece. Of the 2766 patients presenting with onychomycoses (20%), 67% were women and 33% were men. In the toenail infections, dermatophytes were most often isolated (72.3%), especially in women, followed by moulds (9.6%) and yeasts (2%); 16.1% of the infections were mixed. In the fingernail infections mostly yeasts were isolated (72%), especially in women, followed by dermatophytes (10%) and moulds (5.6%); 12.4% of the infections were mixed. PMID- 11856435 TI - Animal models of aspergillosis - also useful for vaccination strategies? AB - The attempt to establish vaccination strategies against infections caused by Aspergillus fumigatus seems to be questionable. Invasive aspergilloses are opportunistic diseases of the immunocompromised host and only a passive immunization with immunoglobulins could be taken into consideration. Until now there have been no preclinical and/or clinical data available concerning the efficacy of specific immunoglobulins; animal experiments could offer an approach for the preclinical assessment of this topic. Generally, A. fumigatus is an opportunistic pathogen. Birds show a relatively high susceptibility to infections caused by A. fumigatus. In laboratory animal species, rabbits seem to have the highest susceptibility followed by mice, rats and guinea-pigs. Mice are easy to handle in all-day laboratory use, and infections are mostly established by the intravenous, intranasal or intraperitoneal route. The main target organs of infection are the kidneys by all three infection routes. Forty clinical isolates of A. fumigatus tested showed a comparable virulence in systemic infections in the intravenously infected mouse model. By using histopathological techniques, we also observed infectious lesions within the central nervous system in all cases. Only A. fumigatus strains lacking green pigmentation showed a significantly lower virulence. Histopathological examinations are of great benefit in the study of these animal models as they give detailed information about the infectious process. Measuring colony-forming units in tissues is only of minor use in prediction as it cannot discriminate between infective tissue lesions and cavity infections/persistence, e.g. in the kidney pelvis. Quantitative methods for measuring fungal organ burdens, e.g. by chitin-enzyme-linked immunosorbent assays, have also been described and offer an alternative towards solely measuring colony-forming units in tissues. PMID- 11856436 TI - Case Report. Multiple etiology post-surgery endophthalmitis. AB - The case describes a septic endophthalmitis arisen in a convalescence period following surgery of cataract extraction. The infection was due to Staphylococcus aureus and three fungal components, Candida albicans, Candida glabrata and Acremonium kiliense, which were subsequently isolated. A careful and prompt laboratory investigation allowed the clinicians to adjust the antimycotic therapy and attain an excellent clinical result. PMID- 11856437 TI - Case report. Fatal rhinocerebral zygomycosis due to Rhizopus oryzae. AB - A case of rhinocerebral zygomycosis due to Rhizopus oryzae, arising after trauma in a 53-year-old diabetic man, is reported. Diagnosis was based on histological and mycological examination. Fragments of the colonies were observed by scanning electron microscopy. This is the first case diagnosed in Tuscany. PMID- 11856438 TI - Case report. Phaeohyphomycosis caused by Phialophora verrucosa developed in a patient with non-HIV acquired immunodeficiency syndrome. AB - A 53-year-old woman had asymptomatic multiple nodules on her gluteal region for 6 months. She had a history of systemic corticosteroid treatment for Evans' syndrome. Recently she had developed an immunodeficiency condition with CD4+ cell depletion without an HIV infection and a normal serum gamma globulin level. A smear from the purulent exudate of the nodules revealed many brown-coloured hyphae, spores and few large dark-brown cells. A short, hairy, dark-brown coloured colony was cultivated on Sabouraud glucose agar. Slide culturing revealed only a Phialophora-type conidia formation, and the fungus was diagnosed as Phialophora verrucosa. Severe immunosuppressive condition (non-HIV acquired immunodeficiency syndrome) of this patients after systemic corticosteroid treatment for Evans' syndrome predisposed an opportunistic cutaneous fungal infection due to P. verrucosa. Cases with cutaneous infection due to P. verrucosa reported in Japan are summarized and discussed. PMID- 11856439 TI - Case report. Tinea corporis due to Microsporum canis in a professional cyclist. AB - We report the case of a 20-year-old man, a professional cyclist, with multiple scaling plaques on his limbs that had been present for 40 days. Our case of tinea corporis presented some peculiarities related to the aetiological agent Microsporum canis and also its mode of transmission. PMID- 11856440 TI - Case report. Monascus purpureus--a new fungus of allergologic relevance. AB - Anaphylactic reactions to food containing allergens in the consumption or preparation of food are well known. However, allergy in the preparation of sausages has rarely been described. In the present study a 26-year-old-butcher was investigated who had a severe anaphylactic reaction developing sneezing, rhinitis, conjunctivitis, generalised pruritus, followed by widespread urticaria, Quincke's oedema and dyspnoe after starting to prepare sausages containing red yield rice. Red yield rice is produced by the fungus Monascus purpureus. It was the first time that Monascus purpureus could be shown as allergic agent by means of prick-to-prick test, Cellular Antigen Stimulation Test (CAST) and different immunoblots. PMID- 11856441 TI - Human Trichuris-specific antibody responses in vaccinated hu-PBL-SCID mice. AB - Trichuris trichiura is a highly prevalent intestinal helminth of humans with a well-characterized animal model, Trichuris muris in the mouse. Relating the murine work back to the human infection has been difficult, however, as many of the questions addressed in the mouse cannot be asked in humans. The ability to reconstitute a mouse with a human immune system could help bridge this gap, allowing a human immune response to be studied under a controllable laboratory environment. In this study, we demonstrate that severe combined immunodeficient mice engrafted with naive human peripheral blood lymphocytes are capable of mounting a Trichuris specific human antibody response after vaccination with T. muris antigens. The phenotype of the response depended on the vaccinating antigen with excretory/secretory antigens eliciting a human immunoglobulin IgG2 response, and whole worm homogenate stimulating IgG1 and IgG2 responses. Vaccination with homogenate also enhanced a human IgG response against a 66-kDa component of T. muris homogenate in a donor-dependent manner. This work shows the potential of using the humanized mouse model for studying the immune responses of humans living in T. trichiura endemic areas. PMID- 11856442 TI - Anti-inflammatory responses and oxidative stress in Nippostrongylus brasiliensis induced pulmonary inflammation. AB - Migration of L3 larvae of Nippostrongylus brasiliensis through the lungs of the rat, during primary infection, was studied at 24 h, 72 h and 8 days. At 24 h p.i., there was evidence of damage to lung epithelial cells and microvasculature, with increased protein and gamma-glutamyl transpeptidase in the bronchoalveolar lavage (BAL) fluid. However, there was little evidence of inflammatory cell recruitment. At 24 h p.i., there was a significant reduction in the inflammatory cytokine tumour necrosis factor alpha. Superoxide (O2-*) production was also reduced, accompanied by an increase in superoxide dismutase activity. Lipid peroxidation was reduced at 24 h p.i. and L3 larvae were shown to possess high levels of glutathione compared to host lung tissue. Nitric oxide, detected as nitrite, was produced in BAL fluid, and inducible nitric oxide synthase protein was increased by 72 h p.i. There was evidence of peroxynitrite production throughout the infection period with specific protein bands nitrosylated at 75, 30 and 25 kDa. It appears that despite early evidence of lung damage, the inflammation was reduced in response to L3 larvae of N. brasiliensis. PMID- 11856443 TI - Heterogeneity in the effect of different ixodid tick species on human natural killer cell activity. AB - Tick saliva plays a vital role in blood-feeding, including manipulation of the host response to tick infestation. Furthermore, a diverse number of tick-borne pathogens are transmitted to vertebrate hosts via tick saliva, some of which exploit the immunomodulatory activities of their vector's saliva. We report that salivary gland extracts (SGE) derived from Dermacentor reticulatus adult ticks induce a decrease in the natural killer (NK) activity of effector cells obtained from healthy human blood donors. The decrease was observed with SGE from both female and male D. reticulatus fed for either 3 or 5 days on mice, but no significant effect was observed with SGE from unfed ticks or ticks that had fed for 1 day. These results indicate that the tick anti-NK factor(s) is only active after blood-feeding has commenced. Microscopic examination revealed that the first step of NK activity, namely effector/target cell conjugate formation, was affected by SGE. The observed reduction in conjugate formation occurred when effector (but not target) cells were treated with SGE for 30 min, and the effect persisted after 12 h of treatment. Similar but less potent anti-NK activity was detected for SGE from Amblyomma variegatum and Haemaphysalis inermis. By contrast, SGE derived from Ixodes ricinus and Rhipicephalus appendiculatus female ticks did not decrease NK activity. The apparent absence of such activity in these two important vectors of tick-borne viruses suggests that control of NK cells does not play an important role in promoting virus transmission, at least for these particular species. PMID- 11856444 TI - Interleukin-4 influences the production of microfilariae in a mouse model of Brugia infection. AB - Sub-cutaneous infection of interleukin (IL)-4-/- mice on the BALB/c background with third stage larva (L3) of Brugia pahangi revealed an altered cytokine profile consistent with the absence of the Th2 promoting cytokine IL-4. Splenocytes from IL-4-/- mice secreted significantly more antigen (Ag)-specific IL-2 and interferon-gamma and significantly less Ag-specific IL-5, compared to those from L3-infected wild-type mice. However, levels of Ag-specific IL-13 were similar between groups. Despite the alteration in immune responses, there was no significant difference in recovery of developing worms from the peritoneal cavity of the two strains of mice at any time postinfection. However, at later time points of infection, the IL-4-/- mice contained large numbers of microfilariae (Mf) in the peritoneal cavity while the wild-type mice contained comparatively few Mf. The differences in Mf levels appear to relate to differences in worm fecundity in the two strains of mice, with adult female worms from the wild-type mice containing few developing Mf. Moreover, implantation of sexually mature adult female worms into the peritoneal cavity of both strains of mice resulted in equal levels of Mf, confirming that the primary role of IL-4 is to limit fecundity during the maturation phase of infection. PMID- 11856445 TI - Mechanisms of immunity to Haemonchus contortus infection in sheep. AB - In two separate experiments, sheep were immunized by nine to 12 weekly immunizing infections with 4000 Haemonchus contortus third stage larva (L3), drenched with anthelminthics and maintained free of H. contortus infection for a further 12 weeks. The anamnestic cellular immune responses in both the abomasal lymph node (ALN) and mucosa of the immunized sheep were examined 3 and 5 days post challenge with 50 000 H. contortus L3. Sheep in the two experiments clearly segregated out in two distinct groups, one in which challenge larvae were obviously present in the tissues of all 12 sheep at 3 and 5 days post challenge while no challenge larvae were detected in tissues of seven of the eight sheep in the other group. In sheep in which no tissue larvae were detected, very few changes were noted in either the ALN or mucosa. In contrast, dramatic changes were observed in the cellular profiles of the ALN and mucosa after challenge infection in sheep in which larvae were observed in the abomasal tissues. In the ALN, these changes were characterized by an increase in the relative percentage of gammadelta-TCR+ T cells and B cells and an increase in the proportion of CD4+ T cells coexpressing the activation markers MHC class II and CD25. In the abomasal mucosa, an increase in the number of infiltrating CD4+ and gammadelta-TCR+ T cells and B cells was observed by 3 days postinfection and these levels were further increased at 5 days postinfection. This infiltration of the abomasal mucosa by lymphocytes was accompanied by a dramatic increase in the number of infiltrating eosinophils, which were often in intimate association with the surface of H. contortus larvae. None of these changes occurred in the mucosa of the sheep that showed no sign of challenge larvae in the tissues; however, a transient increase in gammadelta T cells in the ALN and a drop in intraepithelial globule leucocytes were uniquely observed in these sheep at 5 days post challenge. These results suggest that two different types of immune responses can be generated after challenge infection of immunized sheep, one where tissue larvae are excluded from their tissue niche as observed previously and which is associated with changes in globular leucocyte population but no mobilization of the local immune system. In contrast, when challenge larvae reach their tissue niche, dramatic changes in the local immune system occur, including a pronounced infiltration of eosinophils. These two immune mechanisms may be associated with the rapid and delayed rejection of parasite infections in immune sheep. PMID- 11856446 TI - Serum immunoglobulin E response in calves infected with the lungworm Dictyocaulus viviparus and its correlation with protection. AB - Protection of a primary Dictyocaulus viviparus infection was measured against a homologueous challenge infection in two independent experiments and this was correlated with serum immunoglobulin IgE responses. A primary infection of 30 third stage larvae (L3) of D. viviparus on day 0 protects calves for 70% against a challenge infection of 2000 L3 on day 35 compared to calves with no primary infection. The variation in post mortem worm counts within this group (n = 6) was very large with mean worm counts of 145 (range 3-446) lungworms. Parasite specific IgA, IgE, IgG1 and IgG2 and total IgE levels in serum were measured by ELISA. Parasite specific IgA, IgG1 and IgG2 were elevated after infection, but correlation with protection was only found with IgG1 levels on day 42 and with IgG2 levels on day 70. IgE was measured in a sandwich ELISA using antisheep IgE that cross-reacts with cattle IgE. No parasite specific IgE could be detected. However, total serum IgE was elevated after infection and total serum IgE levels before and on the day of challenge correlated with protection (P < 0.05). Total serum IgE also correlates with peripheral eosinophil counts between days 14 and 28 after primary infection. Western blots with three different parasite antigen preparations, L1, excretory/secretory products and crude worm adult antigens, were used to detect parasite specific IgE in sera depleted of IgG and IgM. These depleted sera from protected calves contained parasite specific IgE, while sera from nonprotected calves were negative. A band of approximately 100 kDa was recognized in all three antigens. In a second experiment, primary doses of 30, 60, 120, 240, 480 and 960 L3 of D. viviparus were used and necropsy was 11 days after challenge. This experiment confirmed the correlation between protection and total IgE levels before and on the day of challenge. The rapid and strong IgE responses in protected animals after such a low infection might be caused by the specific characteristics of the lungworm antigens or by the somatic migration of the worm and might be involved in the rapid development of protection against lungworm reinfections in cattle. PMID- 11856448 TI - 'Acceptable' and 'unacceptable' risks. PMID- 11856449 TI - Response to Dr Silverman's column. 'Acceptable' and 'unacceptable' risks. PMID- 11856450 TI - The roles of the community and physician in treatment decisions for extremely premature infants. PMID- 11856451 TI - Changing patterns of low birthweight and preterm birth in the United States, 1981 98. AB - Low birthweight (LBW) and preterm birth are primary risk factors for infant morbidity and mortality in the US. With increasing multiple births and delayed childbearing, it is important to examine the separate contributions of these characteristics to the increases in LBW and preterm birth rates. US natality records from 1981, 1990 and 1998 were used to calculate LBW (% births <1500, 1500 2499, <2500 g) and preterm (% births <29, 29-32, 33-36, <37 weeks gestation) rates. Data were stratified by maternal race (black or white) and plurality (singleton vs. multiple birth). LBW and preterm rates among singletons were adjusted for maternal age to examine the influence of demographic shifts on LBW trends. From 1981 to 1998, LBW increased 12% among white infants, but remained relatively stable among black infants. During the same time, preterm birth increased 23% among white infants compared with 3% among black infants. For both black and white infants, the increase in LBW and preterm births was greater among multiple births than among singletons. Adjustment for maternal age did not reduce the temporal increase in LBW or preterm birth among singletons. Black infants continue to experience a markedly higher incidence of LBW and preterm birth, but the racial gap in these outcomes has narrowed slightly in recent years as a result of increasing LBW and preterm birth among white births. The differing trends for white and black infants are the consequence of a disparate trend in the incidence and outcome of multiple births coupled with increases in LBW and preterm birth among white singletons. Understanding the differential patterns in birth outcomes among white and black infants is necessary to develop effective interventions designed to decrease racial disparities in pregnancy outcome. PMID- 11856452 TI - Families with a perinatal death: is there an association between the loss and the birthweight of surviving siblings? AB - Our objective was to study birthweight among surviving siblings in families with and without a perinatal loss, and to evaluate whether different causes of death were associated with the results. Data were for 1967-98 from the Norwegian Medical Birth Registry. Births were organised with the mother as the observation unit through the personal identification number, providing sibship files. We analysed 550 930 sibships with at least two singletons, 208 586 sibships with at least three singletons and 45 675 sibships with at least four singleton births. We compared mean birthweight and gestational age between infants in sibships with and without a perinatal loss, total losses and the different causes of death. Surviving siblings in families with a perinatal loss had significantly lower mean birthweights than their counterparts in unaffected families, after adjusting for gestational age, interpregnancy interval, time period and marital status. An exception was found when cause of death was a birth defect, when growth retardation among surviving siblings was not found on average. We conclude that families who have lost an infant because of a birth defect do not appear to have an increased risk of adverse birth outcome associated with growth restriction. PMID- 11856453 TI - Birthweight and gestational age effects on motor and social development. AB - The number of children at risk for delays in motor and social development (MSD) associated with preterm delivery and low birthweight is increasing, but such children are generally not seen as being in need of evaluation. The objective of these analyses was to determine whether there are independent effects of birthweight and gestational age on MSD and the magnitude of effects. Subjects were a representative sample of 4621 US-born singleton children, aged 2-47 months, examined in the third National Health and Nutrition Examination Survey (1988-94). MSD was assessed using an age-appropriate scale. Birthweight and gestational age were taken from birth certificates. Mexican-American and 'other' race/ethnicity (other than non-Hispanic white, non-Hispanic black or Mexican American), low parental education level, older maternal age, higher birth order, low birthweight (LBW, <2500 g) and preterm delivery (<37 weeks) were all found to be associated with significant (P < 0.01) delays in MSD. Three per cent of the infants and children were preterm LBW and 2.2% term LBW (<2500 g, 37-44 weeks). Adjusting for socio-demographic factors, preterm LBW children had lower MSD scores (-1.5 +/- 0.3 points, P < 0.0001) through early childhood, as did term LBW children (0.8 +/- 0.4 points, P < 0.03). For females, LBW was the most important perinatal predictor of a lowered score (-0.9 +/- 0.3 points compared with normal birthweight, P < 0.04). For males, scores were additionally decreased by -0.1 +/- 0.03 points/week (P=0.001) of early delivery. LBW children had less muscle mass, but adjusting for muscularity did not diminish the effects of birth size on MSD. LBW status and preterm delivery are associated independently with small, but measurable, delays in MSD through early childhood and should be considered along with other known risk factors for development delays in determining the need for developmental evaluation. PMID- 11856454 TI - Psychomotor development in Argentinean children aged 0-5 years. AB - In Argentina, there is no information on ages of attainment of developmental milestones and very few data about environmental factors that influence them. A national survey on the psychomotor development of children under 6 years of age was carried out with the help of 129 paediatricians. Logistic regression was applied to a final sample of 3573 healthy, normal children in order to estimate selected centiles (25th, 50th, 75th and 90th), together with their respective confidence intervals, of the ages of attainment of 78 developmental items belonging to the following areas: personal-social (18 items), fine motor (19), language (18) and gross motor (23). The 50th centile obtained for each of the 43 comparable items was compared with those obtained in previously standardised tests: DDST, Denver II, Bayley and Chilean scales. Neither significant nor systematic differences were found between our results and those described in the tests used for comparison. Multiple logistic regressions showed that social class, maternal education and sex (female) were associated with earlier attainment of some selected developmental items, achieved at ages later than 1 year. Selected items achieved before the first year of life were not affected by any of the independent environmental variables studied. The information is useful in helping paediatricians in their daily practice for surveillance of development, as baseline information for epidemiological studies on development in our country and for cross-cultural analysis. PMID- 11856455 TI - An epidemiological study of childhood disability in Jeddah, Saudi Arabia. AB - Childhood disability is a major health problem that is pronounced in developing countries. The objectives of this study were to identify the maternal risk factors present in our society and their impact on acquiring a disabled child. Data were collected from the six governmental rehabilitation institutes and a sample of normal non-disabled school children in Jeddah City. Information on sociodemographic factors and maternal risk factors was collected by in-person interview and copied from the children's institute records. Logistic regression models were fitted to calculate the adjusted odds ratios in order to demonstrate the impact for the various maternal risk factors on having a disabled child. Our results have shown that certain modifiable maternal risk factors affecting child health still exist in our community, such as early and late-age marriage and childbearing, as well as low education, unemployment, multiparity and consanguineous marriages. All these risks were shown to put the mother at higher risk of having an auditory disabled child. Also, older-age childbearing (> or = 30 years), low education level and multiparity were the maternal risk factors identified that influenced the risk of visual disability in children. Similar maternal risk factors, in addition to unemployment, were more observed among mothers of mentally disabled children. A huge effort is required to discourage the potential risk factors in our community that influence child health. PMID- 11856456 TI - Delayed maternity and risk at delivery. AB - We evaluated the effect of the mother's age on the risk of unfavourable pregnancy outcome. The study considered 1348 190 first-born and 957 689 second-born babies born in Italy in 1990-94. The risk of stillbirth, preterm birth in liveborns, and low birthweight in liveborns at term, was first evaluated separately and then globally as a function of maternal age, education and parity. Older (> or = 35 years) mothers were found to run an increased risk, both on single and on global evaluation, with respect to their younger counterparts. Primiparae > or = 35 years of age, of low education, ran a global risk threefold higher (3.14 [3.02, 3.26]) than the young highly educated secondiparae who were the lowest risk (4.64% of babies at risk). From 26 years the global risk rose with each advancing year of maternal age, but parity and education modified the age effect. Linear fitting of the proportion of mothers > or = 35 since 1984 indicates that by the year 2025 about 25% of the mothers may face late child bearing. PMID- 11856457 TI - Abdominal wall defects in Denmark, 1970-89. AB - In the last two to three decades, increasing rates of gastroschisis but not of omphalocele have been reported from different parts of the world. The present study represents a register containing 469 children born with abdominal wall defects based on data retrieved from 20 birth cohorts (1970-89) in three nationwide registries. A tentative estimate of the completeness as regards identification of liveborn and stillborn infants is a minimum of 95% and 90% respectively. All cases were reclassified to 166 cases of gastroschisis, 258 of omphalocele and 16 of gross abdominal wall defect. The average point prevalence at birth of gastroschisis was 1.33 per 10 000 live and stillbirths. During the first decade, an increase in prevalence occurred culminating in 1976, followed by a decrease reaching its initial value in 1983 and then a new increase. Overall, no significant linear trend could be demonstrated for the entire period. The average point prevalence at birth for omphalocele was 2.07 and for gross abdominal wall defect 0.12 per 10 000 live and stillbirths with no significant change in the period. The geographical distribution of gastroschisis and omphalocele showed no difference per county. PMID- 11856458 TI - Prenatal and intrapartum events and sudden infant death syndrome. AB - The purpose of this study was to evaluate specific pregnancy and labour and delivery events that may increase the risk of sudden infant death syndrome (SIDS). A matched case-control study was conducted in five counties in southern California, using California death certificate records. The sample consisted of 239 Caucasian, African-American, Hispanic and Asian mothers of SIDS infants and 239 mothers of control infants matched on sex, race, birth hospital and date of birth. Mothers participated in a detailed telephone interview and provided access to obstetric and paediatric records. More case than control mothers reported a family history of anaemia (OR=2.12, P < 0.001). Placental abruptions were strongly associated with SIDS (unadjusted OR=7.94, [95% CI 1.34,47.12]). There was an increased risk of SIDS death associated with maternal anaemia during pregnancy (OR=2.51, [95% CI 1.25,5.03]), while simultaneously adjusting for maternal smoking during pregnancy, maternal years of education and age, parity, infant birthweight, gestational age, medical conditions at birth, infant sleep position and post-natal smoking. Interactions of anaemia and prenatal smoking as well as anaemia and post-natal smoking were not statistically significant. There were no other statistically significant differences between case and control mothers for pregnancy conditions, labour and delivery events (e.g. caesarean sections, anaesthesia, forceps) or newborn complications (e.g. nuchal cord, meconium aspiration). Anaemia and placental abruptions were significantly associated with an increased risk of SIDS; both are circumstances in which a fetus may become hypoxic, thereby compromising the subsequent growth, development and ultimate survival of the infant. PMID- 11856460 TI - Resuscitation fluids. AB - BACKGROUND AND OBJECTIVES: The aim of this review was to summarize the properties of the commonly available resuscitation fluids and highlight where knowledge of the characteristics of individual fluids might guide their use in clinical practice. Some of the current controversies surrounding resuscitation fluids are also discussed. Blood or blood products, or strategies of fluid administration, are not focused upon. CONCLUSIONS: The clinical trial data does not support the concept of a therapeutic advantage for either crystalloids or colloids, despite claimed theoretical advantages for both classes of fluid. The available colloid solutions have quite distinct profiles of activity, both in terms of their plasma volume expansion profile and other physiological and pharmacological properties. Recent data suggests that physiologically balanced crystalloid and colloid solutions may improve clinical outcomes when compared with saline-based fluids. PMID- 11856461 TI - Effects of solvent/detergent-treated plasma and fresh-frozen plasma on haemostasis and fibrinolysis in complex coagulopathy following open-heart surgery. AB - BACKGROUND AND OBJECTIVES: Solvent/detergent-treated plasma (SDP) contains markedly lower protein S (PS) and plasmin inhibitor (PI) activity than standard fresh-frozen plasma (FFP). It has also been reported that SDP contains no alpha(1)-antitrypsin. Despite the lack of clinical data, it is suspected that SDP may be less effective than FFP in the treatment of complex coagulopathies. We therefore conducted a prospective trial to study the impact of SDP and FFP on haemostasis and fibrinolysis in complex coagulopathy after open-heart surgery. MATERIALS AND METHODS: Patients received either 600 ml of SDP (n = 36) or 600 ml of FFP (n = 31) at an infusion rate of 30 ml/min. The following parameters were measured before treatment and 60 min after termination of plasma infusion: prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen, factor VIII, antithrombin, protein C (PC), free PS and PS activity, prothrombin fragments F1+2 (F1+2), D-dimers (DD), fibrinogen degradation products (FDP), plasmin-plasmin inhibitor complexes (PPI), plasminogen, PI and alpha(1) antitrypsin. RESULTS: The rise in fibrinogen, factor VIII, antithrombin, PC, free PS, alpha(1)-antitrypsin and plasminogen, and the decrease in PT and APTT, did not significantly differ between the two study arms. However, PS activity did not increase after SDP infusion but did show a significant elevation after infusion with FFP. PI declined significantly after SDP and remained uninfluenced by FFP. Neither SDP nor FFP had any significant influence on F1+2, DD or FDP. However, a significant decrease in PPI levels caused by both types of plasma indicated a reduction in hyperfibrinolysis. Clinical haemostasis evaluation revealed no significant difference between the two treatment regimens. No adverse reactions were observed. CONCLUSION: With the exception of PS and PI, SDP and FFP improved haemostasis and fibrinolysis to a similar degree. The clinical significance of these findings has to be determined in patients with severe acquired PS and PI deficiency requiring plasma transfusions. PMID- 11856462 TI - Hepatitis B vaccinees may be mistaken for confirmed hepatitis B surface antigen positive blood donors. AB - BACKGROUND AND OBJECTIVES: Hepatitis B surface antigen (HBsAg) test sensitivities have gradually increased, and neutralizable weak HBsAg-positive donations, with no other hepatitis B virus (HBV) markers, have occasionally been found in our donor population. On investigation, these donors have admitted to receiving hepatitis B vaccine up to 5 days previously. A study was therefore initiated to monitor HBsAg reactivity amongst volunteers after receiving their first dose of hepatitis B vaccine. MATERIALS AND METHODS: Eight volunteers were tested using three HBsAg assays (Abbott Auszyme, Ortho HBsAg-3 and Abbott/Murex GE34/36) on days 0, 3, 5, 7 and 10 after administration of hepatitis B vaccine. RESULTS: Two HBsAg tests (Abbott Auszyme and Ortho HBsAg-3) did not detect HBsAg reactivity amongst the volunteers, although the Abbott Auszyme test results reached 70-80% of the manufacturer's cut-off at day 3 in two volunteers. The most recently launched assay (Abbott/Murex GE 34/36) detected seven (87%) of the eight volunteers as HBsAg reactive on day 3, and two (25%) volunteers were still reactive on day 5. CONCLUSION: The Abbott/Murex GE 34/36 assay demonstrated HBsAg reactivity in most volunteers on day 3 and in some on day 5 after vaccination. It is therefore recommended that individuals who have recently been vaccinated with hepatitis B be deferred from blood donation for at least 7 days. PMID- 11856463 TI - Prevalence of human parvovirus B19 in blood donors as determined by a haemagglutination assay and verified by the polymerase chain reaction. AB - BACKGROUND AND OBJECTIVES: Transmission of human parvovirus B19 (PV B19) by transfusion of blood and blood products is well documented. Although PV B19 infection is connected with severe complications in some recipients, donor screening is not yet mandatory. In this study the prevalence of PV B19, as detected by a haemagglutination assay (the Human PV B19 Antigen-Test), was assessed. In addition, the persistence of B19 DNA and the serological status of blood donors was also assessed. The specificity and utility of the Human PV B19 Antigen-Test for donor screening was investigated and compared with other screening strategies. MATERIALS AND METHODS: The prevalence of PV B19 viraemia was assessed in 28 972 donations from 15,660 remunerated donors by means of the haemagglutination assay. Reactive results were confirmed by the polymerase chain reaction (PCR). RESULTS: Overall, 255 donations gave reactive or indeterminate results in the screening assay. Four donations/donors detected by the haemagglutination assay were confirmed as positive for B19 DNA by PCR. Therefore, a frequency was detected of 1:7243 B19-positive donations and 1:3915 positive donors. Specificity was determined to be 99.1%. Follow-up showed the persistence of viraemia in low concentrations for prolonged time-periods. CONCLUSION: Blood donations with a high level of human PV B19 viraemia can be detected by the haemagglutination assay, which is rapid and easy to perform. The presence of neutralizing antibody may inhibit specific haemagglutination. PMID- 11856464 TI - Establishment of the first World Health Organization International Standard for human parvovirus B19 DNA nucleic acid amplification techniques. AB - BACKGROUND AND OBJECTIVES: A collaborative study, involving 26 laboratories from 14 countries, was carried out in order to establish a World Health Organization (WHO) International Standard for human parvovirus B19 (B19) DNA nucleic acid amplification techniques (NAT). MATERIALS AND METHODS: Four samples: AA, BB (which were lyophilized), CC and DD (which were liquid preparations) were analysed using several different NAT assays. The mean B19 DNA content of each sample was determined for each laboratory using an end-point dilution method. RESULTS: There was good agreement between the overall mean 'equivalents'/ml obtained by the different assays. The mean log(10) 'equivalents'/ml were 5.76 for sample AA, 5.73 for sample BB, 5.82 for sample CC and 7.70 for sample DD. The differences in titre among samples AA, BB and CC were not statistically significant, but the titre of DD was significantly higher. CONCLUSIONS: Despite the range of NAT assays used in the study, it was possible to calculate the mean B19 DNA concentrations in the four preparations. Lyophilized preparation AA was established as the first International Standard for B19 DNA NAT assays and was assigned a concentration of 10(6) international units (IU)/ml. PMID- 11856465 TI - Microbial contamination of cord blood stem cells. AB - BACKGROUND AND OBJECTIVES: After storage, low levels of contaminating bacteria in standard blood components can reach bacteraemic levels, causing severe transfusion-associated sepsis. For cord blood (CB), the significance of low levels of contaminating bacteria and the optimal detection method is unknown and not supported by available guidelines. MATERIALS AND METHODS: Spiking experiments and testing of various subfractions of CB units were used to determine the behaviour of bacteria during centrifugation, freezing and thawing. For routine testing of CB, different volumes were compared for the detection of potential pathogens and micro-organisms of low pathogenicity. RESULTS: Centrifugation, as applied to CB fractionation, does not show concentration of bacteria in any particular fraction and supports the possibility of culture of waste fractions. Dimethylsulphoxide (DMSO) and freezing does not affect the viability of bacteria under the conditions used in this study. Owing to the low contamination level, a large sample volume of 20 ml was more sensitive than a 10-ml sample volume. Eighty five per cent of the isolated strains can be considered to be of low pathogenicity. CONCLUSION: When an optimal waste fraction sample volume of 20 ml was cultured, the contamination rate of CB was found to be approximately 13%, with low levels of < 1 colony-forming unit (CFU)/ml. Such levels of bacteria of low pathogenicity are expected to be of clinical importance only when CB is expanded in vitro for a prolonged period of time. PMID- 11856466 TI - Characterization of the human ABO gene promoter in erythroid cell lineage. AB - BACKGROUND: The human ABO blood group system is important in transfusion and organ transplantation. Although the molecular basis of the ABO gene has been established, recent studies have begun to characterize the mechanism of the ABO gene expression. MATERIALS AND METHODS: Transient transfection assays were carried out in human erythroleukaemia HEL cells and human gastric cancer KATOIII cells. Electrophoretic mobility shift assays were performed using nuclear extracts derived from both cells. RESULTS: Our characterization of the 5' upstream sequence of the ABO genes indicated that the region between -117 and +31 is essential to direct expression of a reporter gene in erythroid cells. We show that a sequence located between positions -22 and -14 of the ABO promoter binds a ubiquitous transcription factor Sp1 or Sp1-like protein(s). Mutation of this site abrogates binding of those factors and reduces the ability of the ABO promoter to function in erythroleukaemia cells and gastric cancer cells. CONCLUSION: The expression of the ABO promoter appears to be influenced by the binding of Sp1 or Sp1-like protein(s) in both erythroid and epithelial cell lineages. PMID- 11856467 TI - Blue-blooded or cherry-blooded: what's in the colour? PMID- 11856469 TI - Regarding 'Including polymerase chain reaction in screening for hepatitis C virus RNA in blood donations is not cost-effective' by Loubiere et al. PMID- 11856471 TI - Standardization of anti-human globulin sera for phenotyping red blood cells using a new method. PMID- 11856473 TI - Helicobacter pylori does not promote N-methyl-N-nitrosourea-induced gastric carcinogenesis in SPF C57BL/6 mice. AB - Helicobacter pylori (H. pylori) infection has been acknowledged as a promoter and an initiator for gastric carcinogenesis in experimental models using Mongolian gerbils with H. pylori strains TN2GF4 and ATCC 43504, which have + ve cagA and vacA phenotype s1 / m1. To get more insight into the role of H. pylori in gastric carcinogenesis, we studied the effect of H. pylori SS1, which has + ve cagA and vacA phenotype s2 / m2, on N-methyl-N-nitrosourea (MNU)-induced chemical gastric carcinogenesis using SPF C57BL / 6 mice. Thus, H. pylori SS1 was inoculated 1 week after the completion of MNU treatment to examine the promoting effect of this bacterium. The incidences of polypoid lesions, differentiated adenocarcinomas, and adenomatous hyperplasias were 67% (10 / 15), 47% (7 / 15) and 80% (12 / 15), respectively, in the MNU-alone group. The corresponding figures were 31% (8 / 26), 23% (6 / 26) and 35% (9 / 26) in the MNU + H. pylori group. The incidences of polypoid lesions and adenomatous hyperplasia were significantly different between the groups. Thus, the results indicate that H. pylori SS1 infection reduced susceptibility to chemical gastric carcinogenesis in this model. The discrepancy between the present result and previous results is likely to have been caused by differences in host factors and bacterial factors. Further study of the relationship between gastric carcinogenesis and H. pylori infection is needed. PMID- 11856474 TI - Uterine adenocarcinoma in N-ethyl-N'-nitro-N-nitrosoguanidine-treated rats with high-dose exposure to p-tert-octylphenol during adulthood. AB - Since many risk factors are associated with the development of uterine adenocarcinomas in humans, the etiology is unclear in most cases, although it has been pointed out that estrogen may play essential roles. To clarify the effects of exposure to p-tert-octylphenol (OP), an environmental xenoestrogen, on uterine carcinogenesis, adult Donryu rats were initiated with a single intrauterine treatment of N-ethyl-N'-nitro-N-nitrosoguanidine (ENNG) at 11 weeks of age and exposed thereafter to 100 mg / kg OP by s.c. injection until 15 months of age. Adult ovariectomized (OVX) rats were also treated in a similar way. OP had no effect on occurrence of persistent estrus in middle age, although uterotrophic effects were obvious in OVX rats. At the termination, development of uterine adenocarcinomas was significantly increased in animals exposed to OP during adulthood. No tumors, but a few focal hyperplasias, developed in OVX rats. These findings suggest that OP has tumor-promoting effects on ENNG-treated endometrium of rats, possibly due to direct action on the uterus, as indicated by the uterotrophic effect when a high dose of OP was given. The results provide clues to the mechanisms of influence of hormonal disrupters on uterine carcinogenesis. PMID- 11856475 TI - Development of hepatocellular adenomas and carcinomas associated with fibrosis in C57BL/6J male mice given a choline-deficient, L-amino acid-defined diet. AB - Development of hepatocellular carcinomas in rats caused by a choline-deficient, L amino acid-defined (CDAA) diet, usually associated with fatty liver, fibrosis, cirrhosis and oxidative DNA damage, has been recognized as a useful model of hepatocarcinogenesis caused by endogenous factors. In the present study, in order to further explore involved factors and genes, we established an equivalent model in spontaneous liver tumor-resistant C57BL/6J mice. Six-week-old males and females were continuously fed the CDAA diet and histological liver lesions and oxidative DNA damage due to 8-hydroxydeoxyguanosine (8-OHdG) were examined after 22, 65 and 84 weeks. In male mice, fatty change and fibrosis were evident at 22 weeks, and preneoplastic foci of altered hepatocytes were seen at an incidence of 8/8 (100%) and a multiplicity of 6.6 +/- 4.0 per mouse at 65 weeks. Hepatocellular adenomas and carcinomas developed at incidences of 16/24 (66.7%) and 5/24 (20.8%), and multiplicities of 1.42 +/- 1.32 and 0.29 +/- 0.62, respectively, at 84 weeks. The female mice exhibited resistance to development of these lesions. The CDAA diet also increased 8-OHdG levels in male but not female mice. These results indicate that a CDAA diet causes hepatocellular preneoplastic foci, adenomas and carcinomas associated with fibrosis and oxidative DNA damage in mice, as in rats, providing a hepatocarcinogenesis model caused by endogenous factors in mice. PMID- 11856476 TI - Dietary conjugated linolenic acid inhibits azoxymethane-induced colonic aberrant crypt foci in rats. AB - The modifying effects of dietary feeding of conjugated linolenic acid (CLN) isolated from the seeds of bitter gourd (Momordica charantia) on the development of azoxymethane (AOM)-induced colonic aberrant crypt foci (ACF) were investigated in male F344 rats to predict its possible cancer chemopreventive efficacy. The effect of CLN on the proliferating cell nuclear antigen (PCNA) index in colonic ACF was also examined. Rats were given subcutaneous injections of AOM (20 mg/kg body weight) once a week for 2 weeks to induce ACF. They also received the experimental diet containing 0.01%, 0.1% or 1% CLN for 5 weeks, starting one week before the first dosing of AOM. AOM exposure produced a substantial number of ACF (108 +/- 21/rat) at the end of the study (week 4). Dietary administration of CLN caused a significant reduction in the frequency of ACF: 87 +/- 14 (19.4% reduction, P < 0.05) at a dose of 0.01%, 69 +/- 28 (36.1% reduction, P < 0.01) at a dose of 0.1% and 40 +/- 6 (63.0% reduction, P < 0.001) at a dose of 1%. Also, CLN administration lowered the PCNA index and induced apoptosis in ACF. These findings might suggest possible chemopreventive activity of CLN in the early phase of colon tumorigenesis through modulation of cryptal cell proliferation activity and/or apoptosis. PMID- 11856477 TI - Chemoprevention of N-methylnitrosourea-induced colon carcinogenesis by ursodeoxycholic acid-5-aminosalicylic acid conjugate in F344 rats. AB - Bile acids enhance colon carcinogenesis in animal models, whereas ursodeoxycholic acid (UDCA) suppresses it. Nonsteroid anti-inflammatory drugs prevent colon cancer development in animals and humans. The aim of the present study was to explore the inhibitory effect of UDCA conjugate with 5-aminosalicylic acid (5 ASA), UDCA-5-ASA conjugate (UDCA-5-ASA), against colon carcinogenesis in rats. One-hundred-and-twenty-nine 7-week-old F344 rats received an intrarectal instillation of 2 mg of N-methylnitrosourea 3 times a week for 3 weeks, and were fed a 0% (control), 0.11% or 0.02% UDCA-5-ASA-, 0.08% UDCA- or 0.03% 5-ASA supplemented diet for the next 27 weeks. The test diets contained an equimolar amount of a test agent, 2.0 mmol/kg diet, except for the 0.02% UDCA-5-ASA diet. The tumor incidence and the mean number of tumors/rat at week 30 were significantly lower and smaller in the UDCA-5-ASA diet groups, 48% and 0.7 in both, and marginally lower in the UDCA and 5-ASA diet groups, 56% and 0.9, and 64% and 0.8, compared to the control group, 83% and 1.3. All the tumors were polypoid in shape, and most of them were differentiated adenocarcinomas restricted to the mucosa or submucosa. An analysis by HPLC for bile acids and 5 ASA in the feces and serum collected at week 30 showed that one-half of ingested UDCA-5-ASA was cleaved into UDCA and 5-ASA in the colon. Thus, the two moieties may have independently affected the promotion stage of carcinogenesis. PMID- 11856478 TI - Overexpression of urokinase-type plasminogen activator in human gastric cancer cell line (AGS) induces tumorigenicity in severe combined immunodeficient mice. AB - The significance of urokinase-type plasminogen activator (uPA) expression in gastric cancer development was tested by using a human uPA cDNA transfection approach and an in vivo severe combined immunodeficient (SCID) mouse model. The AGS gastric cancer cell line, which has urokinase-type plasminogen-activator receptor (uPAR) but lacks uPA, was transfected with a plasmid containing human uPA cDNA and injected into the backs of SCID mice. Compared with the parent AGS cells, uPA protein secretion in AGS-2-, AGS-4-, and AGS-8-transfected cells increased by 26.1-, 34.6-, and 4.8-fold, respectively (P < 0.05). mRNA expression levels of uPA in the AGS-4 clone were much stronger than those in AGS-2 and AGS-8 clones. After the cancer cells (2 x 10(6)) were injected s.c. into the SCID mice, a palpable mass was observed at the injection site at around 140 days post injection, followed by accelerated growth of the xenograft up to 180 days post injection only in the high uPA-producing clone (AGS-4). These results suggest that continuous and high production of uPA by tumor cells is one of the important factors reflecting the malignancy of gastric cancer cells. PMID- 11856479 TI - Overexpression of phosphatidylinositol synthase enhances growth and G1 progression in NIH3T3 cells. AB - Phosphatidylinositol (PI) turnover is thought to play an important role in the regulation of cell growth. PI synthase (PIS, cytidine diphosphate (CDP) diacylglycerol (DG): myo-inositol 3-phosphatidyltransferase, EC 2.7.8.11) acts at the last step in the de novo biosynthesis of PI by catalyzing the condensation of CDP-DG and myo-inositol. To study the physiological role of PIS, we established murine NIH3T3 fibroblasts that stably overexpress PIS, by transfection with PIS cDNA (NIH-PIS cells). In immunofluorescence assays, the constitutively overexpressed PIS was found to be localized in the endoplasmic reticulum, as previously reported for the native enzyme activity. NIH-PIS cells showed an increase in PI synthesis in vitro and in vivo, as well as increased cellular levels of PI-4,5-P2 and PI-3,4,5-P3. They also displayed a decrease in their doubling time and accelerated G1 progression. Overexpression of PIS increased cellular levels of the cyclin D1 and E proteins and Akt kinase activity in serum stimulated quiescent NIH3T3 cells. Moreover, PIS overexpression potentiated the colony formation of NIH3T3 cells in soft agar. These results suggest that PIS accelerates G1 progression and stimulates growth by increasing cellular levels of cyclins D1 and E. PMID- 11856481 TI - Not infrequent K-ras mutations in depressed-type early colorectal carcinomas larger than 10 mm. AB - The aim of this study was to elucidate whether K-ras (codons 12 and 13) mutations occur in depressed-type early colorectal carcinomas (DECas) larger than 10 mm in size. Thirty-four cases of DECas including 27 larger than 10 mm were examined for K-ras mutations by means of microdissection, PCR-RFLP (polymerase chain reaction restriction fragment length polymorphism), and direct sequencing. Although K-ras mutation was infrequent (1/7, 14%) in small (less than 10 mm) DECas, 16/27 (59%) and 17/27 (63%) of DECas larger than 10 mm revealed codon 12, or either codon 12 or 13 mutations, respectively. None of the evaluated pathological factors except size showed a correlation with K-ras mutation. These data indicate that although K-ras mutation could not be involved in the early stage of the progression of DECas, it might play a role at a later stage when the tumor size is over 10 mm. PMID- 11856480 TI - Expression of Hqk encoding a KH RNA binding protein is altered in human glioma. AB - The quaking gene family encodes single KH domain RNA-binding proteins that play vital roles in cell differentiation, proliferation, and apoptotic processes. The human quaking gene, Hqk, maps to 6q25-q26, where cytogenetic alterations associated with a variety of human malignancies, including gliomas have been reported. To assess possible relationships of Hqk with human diseases such as glial tumors, we first isolated the Hqk gene, characterized its structure and expression pattern, and carried out mutational analysis of Hqk in primary tumor samples. The Hqk gene contains 8 exons spanning a approximately 200 kb genomic region, and generating at least four alternatively spliced transcripts, Hqk-5, Hqk-6, Hqk-7 and Hqk-7B, of which Hqk-7 is abundantly expressed in brain. Analysis of primary tumors demonstrated a high incidence of expression alterations of Hqk in gliomas (30%; 6/20), but not in other tumors such as schwannomas (0/3), or meningiomas (0/8). Among the tumor samples showing expression alterations, two were devoid of all three major transcripts, one was missing only the Hqk-5 message, and only the Hqk-7 message was absent in two cases. Our results thus imply the involvement of Hqk in human glial tumor progression. PMID- 11856482 TI - Inactivation of O6-methylguanine-DNA methyltransferase in human lung adenocarcinoma relates to high-grade histology and worse prognosis among smokers. AB - To evaluate the significance of O6-methylguanine-DNA methyltransferase (MGMT) activity in the development of human lung adenocarcinoma (AC), we investigated promoter hypermethylation of the MGMT gene by methylation-specific PCR, and the expression of MGMT protein by immunohistochemistry in relation to smoking history of the patients. In total, 31 of 87 AC patients (35.5%) showed hypermethylation of the MGMT gene, and no significant difference was observed between smokers (37.3%) and non-smokers (33.3%). However, hypermethylation of the MGMT gene increased in parallel with lesser differentiation grade of tumors among smokers (well, 16.7%; moderately, 42.1%; poorly, 57.1%; P = 0.022), although this trend was not observed among non-smokers. Almost all the tumors with promoter hypermethylation of the MGMT gene showed consistently negative MGMT staining by immunohistochemistry. When the prognosis of stage-I patients was compared among smokers, it was apparent that the prognosis of patients with inactivated MGMT was worse than that of MGMT-positive patients (P = 0.036). Such differences in the prognoses were not observed among non-smokers. In conclusion, MGMT inactivation is related to the differentiation grade and the prognosis of lung AC patients among smokers. Although further studies are required, we speculate that smoking may induce hypermethylation, not only of the MGMT gene, but also of other important tumor suppressor genes. PMID- 11856483 TI - Therapeutic and analysis model of intrahepatic metastasis reflects clinical behavior of hepatocellular carcinoma. AB - This study was designed to establish an intrahepatic metastasis model to investigate the biology and therapy of hepatocellular carcinoma (HCC) in mice. A fragment of mouse HCC tumor CBO140C12 was orthotopically implanted into the mouse liver. The number of intrahepatic metastatic colonies and the volume of the implanted tumor increased in a time-dependent manner. At 28 days after fragment implantation, all mice showed intrahepatic metastasis. Intravenous administrations of cisplatin and doxorubicin at 7 and 21 days after the implantation significantly suppressed the growth of the primary tumor nodule, but tended to inhibit intrahepatic metastasis. However, a marked decrease of body weight was observed during the experiment. On the other hand, an inhibitor of matrix metalloproteinases (MMPs), ONO-4817, decreased the gelatinase activity of MMP-9 secreted by CBO140C12 cells, and significantly reduced the number of colonies of intrahepatic metastasis when administered orally. Our established model, which is focused on intrahepatic metastasis, is suitable for evaluating the therapeutic effect of HCC and for analyzing intrahepatic metastasis, because this model reflects the clinical features of HCC and all the steps of tumor metastasis. PMID- 11856485 TI - Bromocriptine reverses P-glycoprotein-mediated multidrug resistance in tumor cells. AB - One of the most important causes of anticancer treatment failure is the development of multidrug resistance (MDR). The main characteristics of tumor cells displaying the MDR phenomena are cross-resistance to structurally unrelated cytotoxic drugs having different mechanisms of action and the overexpression of the MDR1 gene, which encodes a transmembrane glycoprotein named P-glycoprotein (P gp). This study evaluated whether bromocriptine, a D2 dopaminergic receptor agonist, influenced anticancer drug cytotoxicity and P-gp activity in a P-gp expressing cell line compared to a non-expressing subline. The K(i) values for P gp of cyclosporine and verapamil were 1.09 and 540 microM, respectively, and that of bromocriptine was 6.52 microM in a calcein-AM efflux assay using porcine kidney epithelial LLC-PK1 and L-MDR1 cells, overexpressing human P-gp. Bromocriptine at 10 microM reduced the IC50 of doxorubicin (DXR) in K562-DXR from 9000 to 270 ng/ml and that of vincristine (VCR) in K562-VCR from 700 to 0.30 ng/ml, whereas the IC50 values of DXR and VCR in the K562 subline were only marginally affected by these drugs. Bromocriptine restored the anticancer effect of DXR, VCR, vinblastine, vinorelbine and etoposide on MDR-tumor cells overexpressing P-gp. These observations suggest that bromocriptine has the potential to reverse tumor MDR involving the efflux protein P-gp in the clinical situation. PMID- 11856484 TI - Serine proteinase inhibitor 9 can be recognized by cytotoxic T lymphocytes of epithelial cancer patients. AB - Serine proteinase inhibitor 9 (PI-9) inhibits granzyme B-mediated apoptosis and interleukin-1beta-converting enzyme activity. In this study, we report that the PI-9 gene encodes antigenic epitopes recognized by the HLA-A24-restricted and tumor-reactive cytotoxic T lymphocytes (CTLs) of epithelial cancer patients. Screening of an autologous cDNA library using a CTL line recognizing HLA-A24+ tumor cells resulted in the isolation of a cDNA, which had an identical coding region to the previously described PI-9 genes. PI-9 gene was expressed in approximately three-fourths of epithelial cancer cell lines and all leukemic cell lines tested. It was also expressed in normal peripheral blood mononuclear cells (PBMCs), but not in a normal fibroblast cell line. CTL sublines contained T cells capable of recognizing the PI-9(292-300) and PI-9(348-356) peptides among 13 different peptides having the HLA-A24 binding motifs. These two peptides were recognized by the CTL line in a dose-dependent and HLA class-I-restricted manner, and also possessed the ability to induce HLA class I-restricted and tumor reactive CTLs in PBMCs from HLA-A24+ cancer patients. These results demonstrate that PI-9 is recognized by HLA class I-restricted and tumor-reactive CTLs of epithelial cancer patients. PMID- 11856486 TI - Enhanced antitumor effect of ultrasound in the presence of piroxicam in a mouse air pouch model. AB - The antitumor effects of piroxicam, a non-steroidal anti-inflammatory drug, on sarcoma 180 cells under ultrasonic irradiation were examined in a mouse air pouch model. When piroxicam was added to sarcoma 180 suspension under ultrasound irradiation (2 MHz, 10 W, 120 s), the mortality rate of tumor cells immediately after the irradiation and the survival rate of mice were significantly higher than those when ultrasound alone was applied, and these effects of piroxicam were dose-dependent. When D-mannitol was used with piroxicam, the mortality rate of the tumors cells after the irradiation was comparable with that when piroxicam alone was applied, but when L-histidine was used concurrently, the antitumor effect was significantly lower than that when piroxicam alone was applied. Histological examinations one week after the ultrasound irradiation in the presence of piroxicam showed sparse tumor tissue in the air pouch and normal appearance of the air pouch and surrounding tissue. The findings suggest that piroxicam enhances the anti-tumor effects of ultrasound in vivo by increasing the production of singlet oxygen without damage to tissue surrounding the tumor. PMID- 11856487 TI - Antitumor effect of an adeno-associated virus vector containing the human interferon-beta gene on experimental intracranial human glioma. AB - We constructed an adeno-associated virus (AAV) vector containing the human interferon-beta (HuIFN-b ) gene (AAV-IFN-beta) and investigated its antitumor effect against human glioma cells (U251-SP) inoculated into the brain of nude mice. Prior to this, we examined human glioma cells transduced with AAV-IFN-beta using video-enhanced contrast differential interference contrast (VEC-DIC) microscopy. Infection of AAV-IFN-beta induced apoptosis and secondary necrosis in human glioma cells. In in vivo experiments, we confirmed production of HuIFN-beta and induction of heat-shock protein (HSP) in glioma cells transduced with AAV-IFN beta. Growth of the experimental gliomas was completely inhibited by six injections of AAV-IFN-beta, starting 7 days after transplantation of glioma cells. In addition, the survival of mice treated with AAV-IFN-beta was remarkably prolonged. These results indicate that AAV-IFN-beta induces apoptosis of glioma cells and has a strong antitumor effect in this experimental glioma model. PMID- 11856488 TI - [Reduce hepatitis recurrence,improve the survival rate of liver transplantation]. PMID- 11856489 TI - [Orthotopic liver transplantation for end-stage liver diseases in 71 cases]. AB - OBJECTIVE: To assess the role of orthotopic liver transplantation (OLT) in the treatment of end-stage liver diseases and to discuss the experience of the operation. METHODS: Retrospective analysis of clinical data of 71 cases of liver transplantation was performed in our hospital from April 1993 to August 2001. RESULTS: One year survival rate of recipients with benign hepatic disease was over 75%. The survival time and life quality of malignant recipients were also improved. Lamivudine monotherapy during the operation period could reduce HBV reinfection rate. The practice of OLT without veno-venous bypass (VVB) was associated with a shorter operating time, less hemorrhape, and thus less blood transfusion during the operation compared with standard technique of OLT with routine use of VVB. The occurrence rate of biliary complications was 5.98% and vascular complication 8.96%. CONCLUSIONS: OLT should become a routine therapeutic choice for end-stage liver diseases. Lamivudine is helpful to reduce HBV reinfection after OLT in HBV-related liver diseases. OLT without VVB is safe and can be performed in the majority of adult patients. The early diagnosis and timely application of interventional radiological technique are important for the treatment of biliary and vascular complications. PMID- 11856490 TI - [Experience of vascular and bile duct reconstruction in 40 cases of orthotopic liver transplantation]. AB - OBJECTIVE: To summarize the vascular and bile duct reconstruction experience in 40 cases of orthotopic liver transplantation in the southwest hepatobiliary surgery hospital. METHODS: The clinical data of 40 cases of liver transplantation were analyzed retrospectively from Jan. 1999 to Nov. 2001. RESULTS: Mortality rate of this group was 15.0%. Complications included: pulmonary infection (18 cases), MOSF (5 cases), intraperitoneal bleeding (4 cases), ARDS (8 cases), thrombus of hepatic artery (1 case), bile leakage (1 case), and cerebral hemorrhage (1 case). The longest survival time was 31 moths (1 case). There were 15 cases whose survival time was more than 1 year. CONCLUSIONS: The key point of success of liver transplantation relies on excellent vascular and bile duct reconstruction technique. PMID- 11856491 TI - [Prevention and treatment of HBV reinfection following liver transplantation]. AB - OBJECTIVE: To study the prevention and treatment of HBV reinfection after liver transplantation. METHODS: Total 19 cases of chronic fulminant hepatitis B, the end-stage of liver cirrhosis and liver carcinoma complicated with HBV cirrhosis. Were performed liver transplantation and given anti-viral drugs pre and post transplantation. Famciclovir was administered in 4 cases, lamivudine in 13 cases and lamivudine+HBIG in 2 cases. The serum HBVM and liver biopsy immunohistochemistry were performed. RESULTS: Four cases given famciclovir developed reinfection. Serum HBsAg, HBeAg and HBV DNA were positive in 3 cases. Liver biopsy immunohistochemistry showed HBsAg and HBeAg phenotype. Classical viral hepatitis in 1 case occurred, three cases died. In the lamivudin group 7 cases showed positive for HBsAg, 2 cases positive for HBV DNA, 4 cases HBsAg or HBcAg phenotype. One case showed positive for serum anti-HBc the other negative for HBVM, and liver biopsy immunohistochemistry was negative too. CONCLUSIONS: These date suggest that anti-virus prevention and treatment in pre and post liver transplantation with HBV infected correlative disease is necessary, feasible and effective. PMID- 11856493 TI - [Kupffer cells inhibit the experimental liver metastasis from colon cancer]. PMID- 11856492 TI - [Liver transplantation with no-heatbeating donor supplied gaseous oxygen during cold preservation in pigs]. AB - OBJECTIVE: To investigate the availability of liver transplantation with non heart beating (60 min) donors supplied gaseous oxygen by hepatic vein in cold preservation period in pigs. METHODS: Thirty-six pigs were randomly divided into three recipient groups and three donor groups (group A-15 min of warm ischaemia, group B-60 min of warm ischaemia and group C-60 min of warm ischaemia with gaseous oxygen by hepatic vein during cold preservation period). OTL was performed after four hours cold preservation. Postoperative 5 days survival rate of the liver metabolic, function and inflammatory reaction were observed. RESULTS: All animals in group A and C survived after reperfusion for 5 days, but animals in group B died within 3 hours postreperfusion. The damage of liver construction and function were found in group A and group B. There was no significant difference of the changes between group A and C. CONCLUSIONS: Gaseous oxygen persufflation during cold preservation has important protective and resuscitative effect for liver from long-time no-heart beating donors. It was possible to become a promising method for liver transplantation with no hearbeating donors. PMID- 11856494 TI - [Effect of glycogen on calcium of donor liver during ischemia-reperfusion period]. AB - OBJECTIVE: To evaluate the effect of glycogen on calcium concentration of rabbit donor liver during ischemia-reperfusion period. METHODS: Donor group (n=21) was divided into 3 subgroups randomly: Group A (n=7): fasting for 24 hours before harvesting; Group B (n=7): normal laboratory chew; Group C (n=7): normal laboratory chew plus glucose supplement intravenously. Based on the self-created animal model for ischemia-reperfusion, the levels of glycogen content, ATP level, viability of Ca(2+)ATPase and plasmic free Ca(2+) concentration ([Ca(2+)]i) of liver tissue were measured. RESULTS: Before cold preservation, there was a significant difference of glycogen content among the three groups at all time points except at the end of rewarming period. ATP level and Ca(2+)ATPase viability were significantly higher in group C than in other two groups. But the plasmic free Ca(2+) concentration was lower in groups with higher glycogen content. CONCLUSIONS: Donor liver with high glycogen content can provide relatively sufficient ATP, maintain better Ca(2+)ATPase viability and prevent plasmic free Ca(2+) concentration overloading. This maybe an important mechanism for glycogen to ameliorate ischemia-reperfusion injury to the donor livers. PMID- 11856495 TI - [Serum alcohol dehydrogenase activity in alcoholic liver diseases]. PMID- 11856496 TI - [Clone of a novel liver cancer associated gene and analysis of the secondary structures of the predicted protein]. AB - OBJECTIVE: To clone a novel liver cancer associated gene, and to explore the molecular basis of liver cancer genesis. METHODS: Using mRNA differential display polymerase chain reaction (DDPCR) and screening the human placenta cDNA library, we got a full-length cDNA of the gene. We prepared and purified the GST fusion protein and the special polyclonal antibody, engaged in the Western blot and immunohistochemical staining analysis, and analyzed the second structures and predicted the function of the protein by the computer soft. RESULTS: We have got a full-length cDNA of the liver cancer associated gene and identified that the full-length cDNA of the gene could be expressed in 293 eukaryocytes by Western blot assay. We localized the target protein in cytoplasm using the immunohistochemical staining methods, and found two SH3 binding domains and several protein kinase phosphorylation sites by analyzing the second structures. CONCLUSIONS: We have got a novel full-length cDNA of human liver cancer associated gene. PMID- 11856497 TI - [Construction and expression of humanized anti-HBsAg scFv targeting interferon alpha in escherichia coli]. AB - OBJECTIVE: To develop a bacteria expression system to produce the fusion protein of humanized anti-HBsAg scFV and interferon-alpha. METHODS: The expression vector was constructed after cleaving the plasmids harboring the humanized anti-HBsAg scFv and interferon alpha respectively and ligating to linearized pET22b subsequence. The expression of fusion protein in E.coli was analyzed by SDS-PAGE. The binding activity and antiviral activity of the fusion protein was characterized by competing inhibition test and cytopathic effect reduction. RESULTS: The plasmid harboring the in frame arranged fusion gene was constructed and identified. After induction for 12h, a new band close to 4.5 10(4) was observed using SDS-PAGE. Results of competing ELISA and cytopathic effect reduction showed the fusion protein retained its specific binding activity and antiviral activities. CONCLUSIONS: The construction and expression of the fusion gene of humanized anti-HBsAg scFv and interferon in E.coli are successful. PMID- 11856498 TI - [Relation between Beta-2-glycoprotein I and hepatitis B virus surface antigen]. AB - OBJECTIVE: To clarify the binding character between Beta-2-glycoprotein I (Beta-2 GPI) and HBsAg. METHODS: Beta-2-GPI was purified from human plasma and labelled with biotin. Solid phase enzyme linked absorbance assay was used to investigate its binding with HBsAg. RESULTS: Biotinylated Beta-2-GPI was found to bind HBsAg and the reaction could be inhibited by excess unlabelled Beta-2-GPI. CONCLUSIONS: Beta-2-GPI may play a role in hepatitis B virus infection. PMID- 11856499 TI - [Induce treatment of recombinant interferon alpha-1b for chronic hepatitis B and its therapeutic efficiency]. PMID- 11856500 TI - [Replication and transfection of hepatitis B virus DNA into primary duck hepatocytes]. AB - OBJECTIVE: By studying the possibility of obtaining expression of human hepatitis B virus (HBV) genes and production in normal liver cells from heterologous species like normal primary duck hepatocytes (PDH), to investigate the species specificity of HBV infection and replication. METHODS: Two days after transfecting the complete HBV genome into PDH by electroporation (transfected group), HBsAg and HBeAg in the supernatants and lysates of PDH were measured by the IMX system. Meanwhile, replication of HBV in PDH was analyzed by Southern blotting and dot blotting procedures. PDH was electroporated as control. RESULTS: HBsAg in the lysate of transfected group was 9.10 (P/N values, positive?2.1), HBeAg was 1.0 (negative?2.1), both were negative in the supernatants of transfected group. dot blotting revealed that transfected group was strongly positive, whereas the control group was negative. Southern blot analysis of intracellular total DNA indicated that there were relaxed circular (RC), covalently closed circular (CCC) and single-stranded (SS) HBV DNA replicative intermediates in the transfected group, and there was no integrated HBV DNA in the cellular genome. Control groups were negative. CONCLUSIONS: Replication of HBV can occur in hepatocytes from nonmammalian species, which strongly supports the idea that replication of HBV has no critical species-specificity, and yet it depends on the endoenvironment of hepatocyte. PMID- 11856501 TI - [5'NCR Incipient establishment of HCV 5'NCR transgenic mice model]. AB - OBJECTIVE: To evaluate the inhibitory effects of HCV antisense oligonucleotide drugs in vivo. METHODS: Transgenic mice were generated by microinjection. The construct of luciferase controlled by HCV 5'NCR that contains CMV promotor was injected into the male pronuclus of fertilized eggs of ICR mice. RESULTS: Sixty eight survival birth transgenic mice were identified by PCR amplification with tail DNA, 13 of whom were positive with an integration ratio of 19.2% (13/68). Transgenic mRNA was detected by RT-PCR in the tissue of three mice's offspring that contain transgenic DNA. Luciferase expression was detected in a line (35#) using the luciferase assay system and the expression persisted in the F2 generation. The phenotype of the mice in this line was normal and there was no significant difference in physiology from normal mice. CONCLUSIONS: This line of transgenic mice will provide a useful animal model for the study of function of HCV 5'NCR and the evaluation of HCV antisense drugs in vivo. PMID- 11856502 TI - [Dynamic expression of tenascin in rat liver during liver fibrogenesis induced by CCl(4)]. AB - OBJECTIVE: To study the expression of tenascin in normal and fibrotic rat liver. METHODS: Liver fibrosis induced in rat with CCl(4) were divided into three stages: the stage of hepatic injury (4 weeks), early stage of hepatic fibrosis (8 weeks) and later stage of hepatic fibrosis (12 weeks). Tenascin expression in liver tissue was observed by immunohistochemical method and in situ hybridization using digoxigenin-labelled DNA probe. RESULTS: In normal rat liver there was a weak staining for tenascin. In both liver injury stage and early stage of hepatic fibrosis, both mRNA signal and immunostaining for tenascin were significantly increased as compared to that in normal liver. In later stage of hepatic fibrosis, mRNA signal and immunostaining for tenascin were decreased compared with that in early stage of hepatic fibrosis. The cellular source of tenascin in liver mainly restricted in mesenchymal cells. CONCLUSIONS: Tenascin is a component of the extracellular matrix of liver tissue. Plays a role in early matrix organization during liver fibrogenesis. PMID- 11856503 TI - [Study on the natural history of chronic hepatitis B]. AB - OBJECTIVE: By clarifying the natural history of chronic hepatitis B, to evaluate its long-term therapeutic outcome, antiviral drugs efficacy and economic significance. METHODS: A cohort of 183 (mean age of 31.75?.03 years, male/female ratio: 152:31) chronic hepatitis B patients with biopsy-proven and 247 cases of general population as control were followed up by retrospective cohort study. The follow-up time was 11.81?.08 years. This study was focused on long-term clinical outcome including the rate of liver cirrhosis, hepatocellular carcinoma and death, the long-term effect of antiviral drugs and prognostic factors. RESULTS: In chronic hepatitis B patients, 22 (12.02%) developed liver cirrhosis, 12 (6.56%) hepatocellular carcinoma, and 20 (10.93%) died. The cumulative survival probabilities were 97.27%, 91.62%, and 84.47% in 5, 10, and 15 years, respectively. The cumulative probabilities of HCC were 0.00%, 3.19%, and 11.56% in 5, 10, and 15 years, respectively. In 247 control subjects, 6 (2.43%) died, none of them developed cirrhosis or HCC. The rates of death, liver cirrhosis, and HCC in hepatitis B patients were markedly different (P<0.005) compared with controls. The overall mortality of hepatitis B patients was 4.50 folds of the general population. Cox multiple regression analysis showed that old age, severe histological injury, and the positive HBeAg were closely related to liver cirrhosis, while old age, severe histological injury, and male were major factors leading to death. The independent variable of predicted HCC was not found. CONCLUSIONS: The long-term outcome of hepatitis B is poor. PMID- 11856504 TI - [Nucleic acid quantifying assay of hepatitis B virus]. AB - OBJECTIVE: To explore the relation between the nucleic acid content of hepatitis B virus and hepatitis B virus markers, liver function and its pathological state. METHODS: Thirty-nine patients suffering from chronic hepatitis B with positive HBsAg were performed liver puncture for histopathological examination. Fluorescence ration PCR analysis was used for HBV DNA quantifying assessment. Hepatitis B virus markers were detected by ELISA. RESULTS: Serum HBV DNA content was related to hepatitis B virus markers. There was no correlation between HBV DNA level and the Scheuer grade in patients with positive HBsAg, and no obvious correlation between serum glutamic pyruvic transaminase level and HBV DNA content. CONCLUSIONS: There is a significant correlation between HBeAg and HBV DNA, but not between liver inflammation stage and serum HBV DNA content. PMID- 11856506 TI - [Classification of genotyping hepatitis B virus with multiplex PCR]. AB - OBJECTIVE: To establish a simple and practicable method to identify the different genotypes of hepatitis B virus (HBV). METHODS: Based on the alignment of 114 complete nucleotide sequence for HBV DNA of different genotypes, the specific sequence of each genotype was found. Six primer sets were designed for each of the six genotypes according to the genotype-specific sequence, and used separately for PCR. The genotype of HBV was identified according to the positive result of PCR. Three primer sets for B, C and D genotypes were added into a single tube for PCR reaction, and HBV was genotyped according to the length of the amplified DNA. RESULTS: There was no difference in the genotyping result of PCR by single or multiplex primers, which was identical to the PCR-RFLP method. CONCLUSIONS: This multiplex PCR method is simple, precise, sensitive, and easy to use. PMID- 11856505 TI - [Pathologic grading and clinical features of chronic hepatitis B]. AB - OBJECTIVE: To raise the accuracy of clinical diagnosis of chronic hepatitis B. METHODS: The correlation between the clinical features, biochemical tests (the serum total bilirubin-TBil, albumin-ALB, prothrobin activity-PTA, alanine aminotransferase-ALT, albumin/globulin-A/G, and r-globulin-r-G) and histopathological data in 202 patients with chronic hepatitis B (CHB) were studied. RESULTS: Some of presenting symptoms and signs were obviously associated with histological grade and stage. The grade of necroinflammatory activity of CHB was associated with the rising TBil, ALT, GGT and the declining ALB, A/G and PTA. The coincidence of clinical diagnosis and pathology was highest in mild chronic hepatitis, 63.8%~79.0%; then was in marked chronic hepatitis, 40.0%~62.5%. The coincidence was lowest in moderate chronic hepatitis, 10.0%~28.2%. CONCLUSIONS: Great attention should be paid to the significance of symptoms and signs, meanwhile the standard of clinical diagnosis for moderate chronic hepatitis might be relaxed somewhat. PMID- 11856507 TI - [Expression and significance of platelet derived growth factor and its receptor in liver tissues of patients with liver fibrosis]. AB - OBJECTIVE: To study the expression and significance of platelet derived growth factor (PDGF) and its receptor (PDGFR) in liver tissues of patients with chronic hepatitis fibrosis and liver cirrhosis. METHODS: The expression, distribution, quantitation, and correlation of PDGF-A, PDGF-B, PDGFR-alpha, PDGFR-beta, and alpha-SMA in the liver tissues were analyzed by immunohistochemical techniques in 21 patients with chronic hepatitis and 42 patients with liver cirrhosis. RESULTS: In the liver tissues of chronic hepatitis and liver cirrhosis, PDGF and its receptor and alpha-SMA mainly distributed in the fibrotic septa and the infiltration area of inflammation, particularly in branch spindle-shaped cells (activated HSC). The expression of PDGF-B and PDGFR-beta was stronger than that of PDGF-A and PDGFR-alpha with a significant difference between them (P<0.05 approximately 0.01). The expression and distribution of alpha-SMA was basically identical with the expression and distribution of PDGF-A, PDGF-B and PDGFR-alpha, PDGFR-beta and quantitative analysis showed a positive correlation (r=0.606, P<0.001). CONCLUSIONS: PDGF and PDGFR play a key role in liver fibrogenesis and development. The biologic effects of PDGF are elicited through activising HSC. Inhibiting PDGF and its receptor is a new approach to the treatment of liver fibrosis. PMID- 11856508 TI - [Arsenic trioxide for the treatment of medium and advanced primary liver cancer]. PMID- 11856509 TI - [Primary evaluation of the therapeutic effect of non-bioartificial liver support system on severe hepatitis]. PMID- 11856510 TI - [Relationship between clinics and the level of blood HBV DNA in different clinical types of hepatitis B]. PMID- 11856511 TI - [Measurement of serum marks of hepatic fibrosis and portal pressure for the diagnosis of early hepatic fibrosis]. PMID- 11856512 TI - [Effects of octreotide on gastric mucosal massive hemorrhage in patients with portal hypertensive gastropathy]. PMID- 11856513 TI - [TIPSS for the treatment of hepatopulmonary syndrome]. PMID- 11856514 TI - [Alcoholic hepatic and renal lesion and the expression of platelet derived growth factor B chain in the kidney]. PMID- 11856515 TI - [Experimental study of the therapeutic effect of interferon-alpha on liver fibrosis]. PMID- 11856517 TI - [Variations of HBV can affect the reaction of specific CTL]. PMID- 11856516 TI - [Effects of glycyrrhetinic acid and IFN-alpha on HSCs collagen metabolism in rat fibrotic liver of varying stages]. PMID- 11856518 TI - [Hepatic transplantation and HBV reinfection]. PMID- 11856519 TI - [Manufacture and clinical application of FSCLZLY-A ultrafitrating therapeutic equipment for ascites]. PMID- 11856520 TI - [Long-acting interferon: studies on pegylated interferon]. PMID- 11856521 TI - Neural plate patterning by secreted signals. AB - The patterning of the CNS relies on the interaction of multiple signaling molecules such as Sonic Hedgehog, Wnts, and BMPs and their antagonists Chordin and Noggin. The identification of the secreted molecule Tiarin (Tsuda et al., 2002, this issue of Neuron), produced by the nonneural ectoderm at border of the anterior and lateral neural plate, now introduces a novel signaling pathway participating in CNS development. PMID- 11856522 TI - Is synaptic homeostasis just wishful thinking? AB - The bone morphogenetic proteins (BMPs) are secreted polypeptides of the TGF-beta family, whose diverse functions include primary neural induction and the dorsoventral patterning of the neural tube. In this issue of Neuron, Aberle et al. (2002) and Marques et al. present evidence that BMP receptors may also influence the development of synapses. The results suggest a novel mechanism for regulating neuronal growth and synaptic homeostasis during development. PMID- 11856523 TI - Sodium channels: grit, determination, and persistence. AB - Persistent sodium channel activity modulates neuronal gain in a neurotransmitter dependent fashion. Previous studies have suggested that persistent and spike related sodium channel activities are mediated by separate species. In this issue of Neuron, Taddese and Bean (2002) show that a single channel population is sufficient to explain both gating behaviors. A simple allosteric model is provided that can explain the results. PMID- 11856524 TI - Targeted gene repair and its application to neurodegenerative disorders. AB - Synthetic DNA oligonucleotides can direct the exchange of single nucleotides within coding regions of mammalian genes by hybridizing to their complementary sequence in the chromosome and creating a recombination joint structure with a single mismatched base pair. Inherent DNA repair processes recognize the mismatch and resolve it using the DNA sequence of the oligonucleotide vector as the template. This gene surgery approach can be used to repair mutations or to disrupt tri-nucleotide repeats in dysfunctional genes responsible for neurological disorders. PMID- 11856526 TI - A primary culture system for functional analysis of C. elegans neurons and muscle cells. AB - C. elegans has provided important insights into neuromuscular system function and development. However, the animal's small size limits access to individual neurons and muscle cells for physiological, biochemical, and molecular study. We describe here primary culture methods that allow C. elegans embryonic cells to differentiate into neurons and muscle cells in vitro. Morphological, electrophysiological, and GFP reporter studies demonstrate that the differentiation and functional properties of cultured cells are similar to those observed in vivo. Enriched populations of cells expressing specific GFP reporters can be generated by fluorescence-activated cell sorting. Addition of double stranded RNA to the culture medium induces dramatic knockdown of targeted gene expression. Primary nematode cell culture provides a new foundation for a wide variety of experimental opportunities heretofore unavailable in the field. PMID- 11856527 TI - Dorsalization of the neural tube by Xenopus tiarin, a novel patterning factor secreted by the flanking nonneural head ectoderm. AB - We have isolated a novel secreted dorsalizing factor of the neural tube, Xenopus Tiarin, which belongs to the olfactomedin-related family. Tiarin expression starts at the late gastrula stage in the nonneural ectoderm adjacent to the anterior neural plate. Overexpression of Tiarin in the embryo causes expansion of dorsal neural markers and suppression of ventral markers. In the eye-forming field, Tiarin overexpression induces the retinal markers and represses optic stalk markers. Tiarin directly dorsalizes neural tissues in the absence of mesodermal tissues and antagonizes the ventralizing activity of Sonic hedghog (Shh). Unlike BMP4, another dorsalizing factor, Tiarin does not display antineuralizing activity on the ectoderm or mesoderm-ventralizing activity. These findings show that Tiarin is a novel patterning signal candidate acting in the specification of the dorsal neural tube. PMID- 11856528 TI - The Drosophila BMP type II receptor Wishful Thinking regulates neuromuscular synapse morphology and function. AB - Proper synaptic development is critical for establishing all aspects of neural function including learning, memory, and locomotion. Here, we describe the phenotypic consequences of mutations in the wishful thinking (wit) gene, the Drosophila homolog of the vertebrate BMP type II receptor. Mutations in wit result in pharate lethality that can be rescued by expression of a wit transgene in motor neurons but not in muscles. Mutant larvae exhibit small synapses, severe defects in evoked junctional potentials, a lower frequency of spontaneous vesicle release, and an alteration in the ultrastructure of synaptic active zones. These results reveal a novel role for BMP signaling in regulating Drosophila neuromuscular junction synapse assembly and activity and may indicate that similar pathways could govern vertebrate synapse development. PMID- 11856529 TI - wishful thinking encodes a BMP type II receptor that regulates synaptic growth in Drosophila. AB - We conducted a large-scale screen for Drosophila mutants that have structural abnormalities of the larval neuromuscular junction (NMJ). We recovered mutations in wishful thinking (wit), a gene that positively regulates synaptic growth. wit encodes a BMP type II receptor. In wit mutant larvae, the size of the NMJs is greatly reduced relative to the size of the muscles. wit NMJs have reduced evoked excitatory junctional potentials, decreased levels of the synaptic cell adhesion molecule Fasciclin II, and synaptic membrane detachment at active zones. Wit is expressed by a subset of neurons, including motoneurons. The NMJ phenotype is specifically rescued by transgenic expression of Wit only in motoneurons. Thus, Wit appears to function as a presynaptic receptor that regulates synaptic size at the Drosophila NMJ. PMID- 11856530 TI - Drosophila Dscam is required for divergent segregation of sister branches and suppresses ectopic bifurcation of axons. AB - Axon bifurcation results in the formation of sister branches, and divergent segregation of the sister branches is essential for efficient innervation of multiple targets. From a genetic mosaic screen, we find that a lethal mutation in the Drosophila Down syndrome cell adhesion molecule (Dscam) specifically perturbs segregation of axonal branches in the mushroom bodies. Single axon analysis further reveals that Dscam mutant axons generate additional branches, which randomly segregate among the available targets. Moreover, when only one target remains, branching is suppressed in wild-type axons while Dscam mutant axons still form multiple branches at the original bifurcation point. Taken together, we conclude that Dscam controls axon branching and guidance such that a neuron can innervate multiple targets with minimal branching. PMID- 11856532 TI - Subthreshold sodium current from rapidly inactivating sodium channels drives spontaneous firing of tuberomammillary neurons. AB - A role for "persistent," subthreshold, TTX-sensitive sodium current in driving the pacemaking of many central neurons has been proposed, but this has been impossible to test pharmacologically. Using isolated tuberomammillary neurons, we assessed the role of subthreshold sodium current in pacemaking by performing voltage-clamp experiments using a cell's own pacemaking cycle as voltage command. TTX-sensitive sodium current flows throughout the pacemaking cycle, even at voltages as negative as -70 mV, and this current is sufficient to drive spontaneous firing. When sodium channels underlying transient current were driven into slow inactivation by rapid stimulation, persistent current decreased in parallel, suggesting that persistent sodium current originates from subthreshold gating of the same sodium channels that underlie the phasic sodium current. This behavior of sodium channels may endow all neurons with an intrinsic propensity for rhythmic, spontaneous firing. PMID- 11856531 TI - Rescue of ataxia and preplate splitting by ectopic expression of Reelin in reeler mice. AB - The gene mutated in reeler (reelin) encodes a protein secreted by neurons in the developing brain that controls laminar positioning of migrating cells in the CNS by an unknown mechanism. To investigate Reelin function, we used the nestin promoter to express Reelin ectopically in the ventricular zone and other brain regions in transgenic mice. In the presence of the endogenous protein, ectopic Reelin did not alter cell migration in the neocortex or the cerebellum. However, in the reeler background, ectopic Reelin induced tyrosine phosphorylation of Dab 1 in the ventricular zone and rescued some, but not all, of the neuroanatomic and behavioral abnormalities characteristic of reeler. These results indicate that Reelin does not function simply as a positional signal. Rather, it appears to participate in multiple events critical for neuronal migration and cell positioning. PMID- 11856533 TI - Temporal synaptic tagging by I(h) activation and actin: involvement in long-term facilitation and cAMP-induced synaptic enhancement. AB - Presynaptic I(h) channels become activated during a tetanus through membrane hyperpolarization resulting from Na(+) accumulation and electrogenic Na(+)/K(+) exchange. I(h) activation is obligatory for inducing long-term facilitation (LTF), a long-lasting synaptic strengthening. cAMP-induced synaptic enhancement also requires I(h) activation, and both processes are sensitive to actin depolymerization. Other mechanisms are responsible for expression of the responses. Once initiated, continued response to cAMP is I(h) and actin independent. Moreover, LTF-induced activation of I(h) renders subsequent cAMP enhancement insensitive to both I(h) blockers and actin depolymerization. This actin-stabilized "temporal synaptic tagging" set by I(h) activation is prolonged when I(h) is activated concurrent with an elevation in presynaptic calcium concentration ([Ca(2+)]i), permitting the further strengthening of synapses given appropriate additional stimuli. PMID- 11856534 TI - Muscarinic induction of hippocampal gamma oscillations requires coupling of the M1 receptor to two mixed cation currents. AB - Oscillatory network activity at gamma frequencies is assumed to be of major importance in cortical information processing. Whereas the synaptic mechanisms of gamma oscillations have been studied in detail, the ionic currents involved at the cellular level remain to be elucidated. Here we show that in vitro gamma oscillations induced by muscarine require activation of M1 receptors on hippocampal CA3 pyramidal neurons and are absent in M1 receptor-deficient mice. M1 receptor activation depolarizes pyramidal neurons by increasing the mixed Na(+)/K(+) current I(h) and the Ca(2+)-dependent nonspecific cation current I(cat), but not by modulation of I(M). Our data provide important insight into the molecular basis of gamma oscillations by unequivocally establishing a novel role for muscarinic modulation of I(h) and I(cat) in rhythmic network activity. PMID- 11856535 TI - Tonic and spillover inhibition of granule cells control information flow through cerebellar cortex. AB - We show that information flow through the adult cerebellar cortex, from the mossy fiber input to the Purkinje cell output, is controlled by furosemide-sensitive, diazepam- and neurosteroid-insensitive GABA(A) receptors on granule cells, which are activated both tonically and by GABA spillover from synaptic release sites. Tonic activation of these receptors contributes a 3-fold larger mean inhibitory conductance than GABA released synaptically by high-frequency stimulation. Tonic and spillover inhibition reduce the fraction of granule cells activated by mossy fiber input, generating an increase of coding sparseness, which is predicted to improve the information storage capacity of the cerebellum. PMID- 11856536 TI - Three-dimensional shape representation in monkey cortex. AB - Using fMRI in anesthetized monkeys, this study investigates how the primate visual system constructs representations of three-dimensional (3D) shape from a variety of cues. Computer-generated 3D objects defined by shading, random dots, texture elements, or silhouettes were presented either statically or dynamically (rotating). Results suggest that 3D shape representations are highly localized, although widely distributed, in occipital, temporal, parietal, and frontal cortices and may involve common brain regions regardless of shape cue. This distributed network of areas cuts across both "what" and "where" processing streams, reflecting multiple uses for 3D shape representation in perception, recognition, and action. PMID- 11856537 TI - Fear conditioning in humans: the influence of awareness and autonomic arousal on functional neuroanatomy. AB - The degree to which perceptual awareness of threat stimuli and bodily states of arousal modulates neural activity associated with fear conditioning is unknown. We used functional magnetic neuroimaging (fMRI) to study healthy subjects and patients with peripheral autonomic denervation to examine how the expression of conditioning-related activity is modulated by stimulus awareness and autonomic arousal. In controls, enhanced amygdala activity was evident during conditioning to both "seen" (unmasked) and "unseen" (backward masked) stimuli, whereas insula activity was modulated by perceptual awareness of a threat stimulus. Absent peripheral autonomic arousal, in patients with autonomic denervation, was associated with decreased conditioning-related activity in insula and amygdala. The findings indicate that the expression of conditioning-related neural activity is modulated by both awareness and representations of bodily states of autonomic arousal. PMID- 11856538 TI - Measurement of steroid sex hormones in serum: a comparison of radioimmunoassay and mass spectrometry. AB - Concern has been raised about the adequacy of radioimmunoassays to measure steroid sex hormones in population studies. We compared steroid sex hormone measurements in serum by radioimmunoassay with mass spectrometry. Four male and four female serum pools with known relative concentrations of steroid sex hormones were measured multiple times by both methods. Because measurements are expected to increase linearly with concentration for each sex, we examined whether the linear regressions of hormone measurements on concentration were the same for radioimmunoassay and mass spectrometry. Estradiol, estrone, androstenedione, testosterone, and dehydroepiandrosterone sulfate were measured in female pools; testosterone, dihydrotestosterone, androstenedione, and dehydroepiandrosterone sulfate were measured in male pools. Regression slopes for radioimmunoassay and mass spectrometry measurements were comparable for all hormones except androstenedione, which had a steeper slope when measured by mass spectrometry (P < or = 0.02). Intercepts for radioimmunoassay and mass spectrometry were similar and close to zero for estradiol, androstenedione, dehydroepiandrosterone sulfate, and in male samples, testosterone. For testosterone in female samples, estrone, and dihydrotestosterone, radioimmunoassay and mass spectrometry intercepts differed significantly. Standard deviations of individual measurements by radioimmunoassay and mass spectrometry differed by hormone and serum concentration; neither method consistently measured hormone concentrations with less variability. Our findings suggest that although absolute concentrations may differ for some hormones, radioimmunoassay and mass spectrometry can yield similar estimates of between subject differences in serum concentrations of most steroid sex hormones commonly measured in population studies. Relative power of studies using radioimmunoassay and mass spectrometry will depend on the hormones measured and their serum concentrations. PMID- 11856539 TI - Spirostanic analogues of castasterone. AB - The synthesis, spectroscopic characterization, and biologic activity of two bioactive spirostanic analogues of the naturally occurring brassinosteroid castasterone are described. PMID- 11856540 TI - Norethindrone acetate (NA) and ethinyl estradiol (EE) related oxidative transformation products in stability samples of formulated drug product: synthesis of authentic references. AB - Preparative chemical methods for the synthesis of eight oxidative transformation products of ethinyl estradiol (EE) and norethindrone acetate (NA) are described. The prepared materials are useful as reference materials and standards for pharmaceutical analysis of EE and NA as bulk chemical or in formulated product. All eight products result from oxidation of the A and/or B rings of the parent compounds. Oxidation of the heteroannular 3,5 dienyl acetate derivative of NA resulted in the 6 alpha-hydroxy, 6 beta-hydroxy and 6-keto NA. Oxidation of 6 keto NA led to the preparation of 6 alpha-hydroxy, 6 beta-hydroxy, 6-keto- and Delta(6) EE. Delta(11) EE was prepared from estrone. PMID- 11856541 TI - A highly specific heterologous enzyme immunoassay for 5 alpha-androstane-3 alpha, 17 beta-diol 17-glucuronide (androstanediol-17G) and developmental patterns of urinary androstanediol-17G excretions. AB - We established a highly specific enzyme immunoassay (EIA) for 5 alpha-androstane 3 alpha, 17 beta-diol 17-glucuronide (androstanediol-17G). Rabbit antisera raised against 5 alpha-androstane-3 alpha, 11 alpha, 17 beta-triol 17-glucuronide 11 glutaryl bovine serum albumin and a heterologous tracer of androstanediol-17G conjugated with horseradish peroxidase at the glucuronic acid group were used. The EIA showed excellent specificity: there were no remarkable cross-reactivities with related androgens. The assay range for urine samples was 0.3-30 ng/ml. Recoveries of standards added to samples were 100-108%. Intra-assay and inter assay coefficients of variation were 2.9-4.4% and 5.7-7.9%, respectively. The EIA was applied to urine samples of 407 males and 322 females to determine developmental patterns and normal ranges of androstanediol-17G excretions in 11 age groups (0 y, 1 y, 2-3 y, 4-5 y, 6-7 y, 8-9 y, 10-11 y, 12-13 y, 14-15 y, 16 17 y, and over 18 y). Urinary androstanediol-17G/creatinine (androstanediol 17G/Cre) ratios in both sexes were high in infancy, tended to decrease during childhood, and began to increase near adolescence. While androstanediol-17G/Cre ratio in girls increased at 8-9 y and reached a plateau during adolescence, that in boys increased at 10-11 y and continued to increase throughout adolescence. Androstanediol-17G/Cre ratios in girls were higher than those in boys at 6-7 y (P < 0.05) and at 8-9 y (P < 0.01). Androstanediol-17G/Cre ratios in boys were higher than those in girls at 12-13 y and at older ages (P < 0.01). These developmental patterns are parallel to age-related changes in androgenicity and serum androstanediol-17G, suggesting that urinary androstanediol-17G/Cre ratio could be a good marker for androgenicity in childhood. PMID- 11856542 TI - Time-dependent aromatase inactivation by 4 beta,5 beta-epoxides of the natural substrate androstenedione and its 19-oxygenated analogs. AB - Aromatase catalyzes the conversion of androgens to estrogens through three sequential oxygenations. To gain insight into the catalytic function of aromatase and its aromatization mechanism, we studied the inhibition of human placental aromatase by 4 beta,5 beta-epoxyandrostenedione (5) as well as its 19-hydroxy and 19-oxo derivatives (6 and 7, respectively), and we also examined the biochemical aromatization of these steroids. All of the epoxides were weak competitive inhibitors of aromatase with apparent K(i) values ranging from 5.0 microM to 30 microM. The 19-methyl and 19-oxo compounds 5 and 7 inactivated aromatase in a time-dependent manner with k(inact) of 0.048 and 0.110 min(-1), respectively, in the presence of NADPH. In the absence of NADPH, only the former inhibited aromatase with a k(inact) of 0.091 min(-1). However, 19-hydroxy steroid 6 did not cause irreversible inactivation either in the presence or absence of NADPH. Gas chromatography-mass spectrometric analysis of the metabolite produced by a 5-min incubation of the three epoxides with human placental microsomes in the presence of NADPH under air revealed that all three compounds were aromatized to produce estradiol with rates of 8.82, 0.51, and 1.62 pmol/min/mg protein for 5, 6, and 7, respectively. In each case, the aromatization was efficiently prevented by 19 hydroxyandrost-4-en-17-one, a potent aromatase inhibitor. On the basis of the aromatization and inactivation results, it seems likely that the two pathways, aromatization and inactivation, may proceed, in part, through a common intermediate, 19-oxo compound 7, although they may be principally different. PMID- 11856543 TI - 11 beta-Hydroxysteroid dehydrogenase antisense affects vascular contractile response and glucocorticoid metabolism. AB - Glucocorticoids (GC's) are metabolized in vascular tissue by two isoforms of 11 beta-hydroxysteroid dehydrogenase (11 beta-HSD). 11 beta-HSD2 is unidirectional and metabolizes GC's to their respective inactive 11-dehydro derivatives. 11 beta HSD1 is bi-directional, also possessing reductase activity and thus the ability to regenerate active GC from the 11-dehydro derivatives. In vascular tissue, GC's amplify the pressor responses to catecholamines and angiotensin II and may down regulate certain depressor systems such as nitric oxide and prostaglandins. We hypothesize that both 11 beta-HSD2 and 11 beta-HSD1 regulate GC levels in vascular tissue and are part of additional mechanisms that control vascular tone. We examined the effects of specific antisense oligomers to 11 beta-HSD2 and 11 beta-HSD1 on GC metabolism and contractile response to phenylephrine (PE) in rat aortic rings. In aortic rings incubated (24 h) with corticosterone (B) (10 nmol/l) and 11 beta-HSD2 antisense (3 micromol/l), the contractile response to graded concentrations of PE (PE: 10 nmol/l - 1 micromol/l) were significantly (P < 0.05) increased compared to rings incubated with B and 11 beta-HSD2 nonsense. 11 beta-HSD1 antisense oligomers also enhanced the ability of B to amplify the contractile response to PE. In addition, 11 beta-HSD2 and 11 beta-HSD1 antisense also decreased the metabolism of B to 11-dehydro-B. 11-Dehydro-B (100 nmol/l) also amplified the contractile response to PE in aortic rings (P < 0.01), most likely due to the generation of active corticosterone by 11 beta-HSD1-reductase; this effect was significantly attenuated by 11 beta-HSD1 antisense. 11 beta-HSD1 antisense also caused a marked decrease in the metabolism of 11-dehydro-B back to B by 11 beta-HSD1-reductase. These findings underscore the importance of 11 beta HSD2 and 11 beta-HSD1 in regulating local concentrations of GC's in vascular tissue. They also indicate that decreased 11 beta-HSD2 activity may be a possible mechanism in hypertension and that 11 beta-HSD1-reductase may be a possible target for anti-hypertensive therapy. PMID- 11856544 TI - Synthesis of some A- and D-ring fused steroidal pyrazoles, isoxazoles and pyrimidines. AB - The preparation of steroidal heterocycles containing pyrazole, isoxazole and pyrimidine rings fused to the 2,3- and 16,17-positions of the steroid nucleus is described. These were prepared by the reaction of hydrazine, hydroxylamine and guanidine, respectively, with 2-ethoxymethylene-3-oxo- or 16-ethoxymethylene-17 oxo- or 2-bis(methylthio)methylene-3-oxo- or 16-bis(methylthio)methylene-17-oxo steroids. PMID- 11856545 TI - New steroidal antiinflammatory antedrugs: Methyl 3,20-dioxo-9 alpha-fluoro-11 beta,17 alpha,21-trihydroxy-1,4-pregnadiene-16 alpha-carboxylate and its 21-O acyl derivatives. AB - In a continuing effort to increase local to systemic activity ratios of potent steroidal antiinflammatory antedrugs, a series of 21-O-acyl derivatives of methyl 3,20-dioxo-9 alpha-fluoro-11 beta,17 alpha,21-trihydroxy-1,4-pregnadiene-16 alpha carboxylate, FP16CM, were synthesized. These derivatives were evaluated for antiinflammatory activity and their adverse effects in an acute and semi-chronic croton oil-induced ear edema bioassay. Following a single topical application in the croton oil-induced ear edema bioassay, treatment with all the compounds resulted in dose-dependent inhibition of edema. From these dose-response profiles, the following ID(50) values (nmol/ear resulting in a 50% reduction of edema) were calculated: prednisolone (Pred); 454, FP16CM; 255, 21-acetate (FP16CM acetyl); 402, 21-propionate (FP16CM-propionyl); 474, 21-valerate (FP16CM valeryl); 446 and 21-pivalate (FP16CM-pivalyl); 219 nmol. In a 5-day semi-chronic study at the equipotent doses, the novel steroidal antedrugs did not significantly alter body weight gain, thymus weights or plasma corticosterone levels unlike the parent compound Pred. The compounds were assessed for high affinity glucocorticoid receptor binding and glucocorticoid-mediated inhibition of nitric oxide (NO) generation in an in vitro RAW 264.7 macrophage cell culture system. Binding affinities for cytosolic glucocorticoid receptors were Pred; 85, FP16CM-acetyl; 86, FP16CM-propionyl; 169, FP16CM-valeryl; 149, FP16CM-pivalyl; 126 nM, respectively. Concomitant potencies for inhibition of NO generation by macrophages stimulated with lipopolysaccharide were Pred; 159, FP16CM-acetyl; 377, FP16CM-propionyl; 405, FP16CM-valeryl; 344, FP16CM-pivalyl; 311 nM, respectively. Collectively, results of these investigations suggest that esterification of 21-OH with various anhydrides did not improve receptor binding, inhibition of NO generation and ear edema inhibition, however, serum corticosterone level and local over systemic activities (L/S) were markedly improved. PMID- 11856546 TI - Ergosteroids V: preparation and biological activity of various D-ring derivatives in the 7-oxo-dehydroepiandrosterone series. AB - Our previous finding that D-ring seco derivatives of dehydroepiandrosterone retained biologic activity (Reich et al., Steroids 1998;63:542-53) motivated us to synthesize and test a number of steroids in which the D-ring is retained but altered in various ways. Several new steroids were synthesized and characterized by (1)H and (13)C NMR spectroscopy. The availability of a number of closely related compounds allowed detailed (13)C chemical shift correlations. Using the induction of two thermogenic enzymes in rats, liver mitochondrial glycerophosphate dehydrogenase (GPDH) and cytosolic malic enzyme, as criteria of biologic activity some 30 compounds were assayed. Hydroxylation of dehydroepiandrosterone (DHEA) at the 16 alpha position was previously shown to diminish activity (Lardy et al., Steroids 1998;63:158-65); the corresponding 7 oxo compound is fully active. Hydroxylation at the 15 beta position of DHEA, 7 oxo-DHEA, or 16 alpha-hydroxy-7-oxo-DHEA greatly diminished the induction of GPDH but induction of malic enzyme was retained. Most 5,15 diene steroids tested had 2 weak, or no, ability to enhance the formation of GPDH but did increase malic enzyme. PMID- 11856547 TI - Evidence for distinct membrane receptors for 1 alpha,25-(OH)(2)D(3) and 24R,25 (OH)(2)D(3) in osteoblasts. AB - 1 alpha,25-(OH)(2)D(3) exerts its effects on chondrocytes and enterocytes via nuclear receptors (1,25-nVDR) and a separate membrane receptor (1,25-mVDR) that activates protein kinase C (PKC). 24R,25-(OH)(2)D(3) also stimulates PKC in chondrocytes, but through other membrane mechanisms. This study examined the hypothesis that osteoblasts possess distinct membrane receptors for 1 alpha,25 (OH)(2)D(3) and 24R,25-(OH)(2)D(3) that are involved in the activation of PKC and that receptor expression varies as a function of cell maturation state. 1 alpha,25-(OH)(2)D(3) stimulated PKC in well differentiated (UMR-106, MC-3T3-E1) and moderately differentiated (ROS 17/2.8) osteoblast-like cells, and in cultures of fetal rat calvarial (FRC) cells and 2T3 cells treated with rhBMP-2 to promote differentiation. 24R,25-(OH)(2)D(3) stimulated PKC in FRC and 2T3 cultures that had not been treated to induce differentiation, and in ROS 17/2.8 cells. MG63 cells, a relatively undifferentiated osteoblast-like cell line, had no response to either metabolite. Ab99, a polyclonal antibody generated to the chick enterocyte 1,25-mVDR, but not a specific antibody to the 1,25-nVDR, inhibited response to 1 alpha,25-(OH)(2)D(3). 1 alpha,25-(OH)(2)D(3) exhibited specific binding to plasma membrane preparations from cells demonstrating a PKC response to this metabolite that is typical of positive cooperativity. Western blots of these membrane proteins reacted with Ab99, and the Ab99-positive protein had an Mr of 64 kDa. There was no cross-reaction with antibodies to the C- or N-terminus of annexin II. The effect of 24,25-(OH)(2)D(3) on PKC was stereospecific; 24S,25 (OH)(2)D(3) had no effect. These results demonstrate that response to 1 alpha,25 (OH)(2)D(3) and 24R,25-(OH)(2)D(3) depends on osteoblast maturation state and suggest that specific and distinct membrane receptors are involved. PMID- 11856548 TI - New highly calcemic 1 alpha,25-dihydroxy-19-norvitamin D(3) compounds with modified side chain: 26,27-dihomo- and 26,27-dimethylene analogs in 20S-series. AB - New highly potent 2-substituted (20S)-1 alpha,25-dihydroxy-19-norvitamin D(3) analogs with elongated side chain were prepared by Wittig-Horner coupling of A ring phosphine oxide with the corresponding protected (20S)-25-hydroxy Grundmann's ketones. Biologic evaluation in vitro and in vivo of the synthesized compounds was accomplished. All the synthesized vitamins possessing a 25 hydroxylated saturated side chain were slightly less active (3-5X) than 1 alpha,25-dihydroxyvitamin D(3) in binding to the porcine intestinal vitamin D receptor and significantly more potent (12-150X) in causing differentiation of HL 60 cells. In vivo, 2-methylene-26,27-dihomo and 2 alpha-methyl-26,27-dimethylene analogs were at least 10 times more active, and 2 alpha-methyl-26,27-dihomo compound at least 5 times more active than the vitamin D hormone both in stimulating intestinal calcium transport and bone calcium mobilization (serum calcium increase). It was also established that a 260 pmol dose of the corresponding 2 beta-methyl analogs had a similar effect on intestinal calcium transport and a much more pronounced effect on bone calcium mobilization as the same dose of 1 alpha,25-dihydroxyvitamin D(3). PMID- 11856549 TI - Characterization of rat liver bile acid acyl glucuronosyltransferase. AB - Recent studies have suggested that bile acid acyl glucuronides form covalently bound protein adducts which may cause hypersensitivity reactions and increased morbidity in patients. Although the preferential biosynthesis of the acyl glucuronides has been known, the characterization of hepatic bile acid acyl glucuronosyltransferase has not yet been clearly elucidated. We have investigated the substrate specificity of the hepatic bile acid acyl glucuronosyltransferase using five common bile acids as substrates. The glucuronidation rate was dependent on the number of the hydroxy group on the steroid nucleus and mono hydroxylated lithocholic acid, the more lipophilic common bile acid, was most effectively metabolized into its acyl glucuronide. The tri-hydroxylated cholic acid, the more water-soluble common bile acid, barely transformed into its glucuronide. Results showed decreasing of the initial velocity of the acyl glucuronidation with increasing of the concentration of substrate, lithocholic acid, owing to the substrate inhibition of the hepatic bile acid acyl glucuronosyltransferase. The substrate analogues, glycine and taurine conjugated bile acids, which exist in the body fluids in high concentrations, also inhibited the enzyme's activity. In addition, enzymatic reaction products, bile acid acyl glucuronides, also inhibited the activity. These inhibitory mechanisms may be responsible for the low concentration of bile acid acyl glucuronides in urine and may be an important detoxification system in the body. PMID- 11856550 TI - Nucleosteroids: carbocyclic nucleoside analogs of androst-4-en-17 beta-ol. AB - Steroidal nucleoside analogs were synthesized starting from testosterone. By reduction of the oxime of 17 beta-hydroxy-androst-4-en-3-one (testosterone), a mixture of the two amino epimers of C-3 were obtained. The 3 alpha-amino-androst 4-en-17 beta-ol was crystallized in 73% yield and coupled with 5-amino-4,6 dichloropyrimidine to give 3 alpha-(5'-amino-4'-chloro-pyrimidin-6'-yl)amino androst-4-en-17 beta-ol. This compound was treated with triethyl orthoformate in acid media to give the corresponding purinyl steroid adduct 3 alpha-(6'-chloro purin-9'-yl)-androst-4-en-17 beta-ol in 98% yield. This substance, in turn, was converted with good yield into the 6'-thio, 6'-methylamino, and 6'-diethyl aminopurinyl derivatives through nucleophilic reactions at C-6 of the purine nucleus. PMID- 11856551 TI - A relatively simple and rapid multi-component method for analysis of steroid profiles in blood, fecal and liver samples. AB - Various methods of steroid analysis were assessed using radiolabeled steroids and thin layer chromatography. Three reversed phase chromatography systems were evaluated for separation and recovery of steroids extracted from blood, liver tissue and feces. The use of different numbers of Sep-Pak C(18) cartridges for the purification of steroid extracts was examined and steroid recoveries were measured and compared. The results indicated that recoveries were best when 4-6 cartridges were used. Rapid and slow procedures of enzymatic hydrolysis and acidic solvolysis of steroid conjugates were compared. A new and relatively rapid method for analysis of steroid profiles in liver, blood and fecal samples was developed. Assessment of this method showed that steroid recoveries were improved compared to existing methods with percentage recoveries of 64.1-82.5 for liver samples, 55.2-75 for blood samples, and 65.1-76.3 for fecal samples. PMID- 11856552 TI - Changes of ovarian interstitial cell hormone receptors and behavior of resident mesenchymal cells in developing and adult rats with steroid-induced sterility. AB - In the present paper, we report that injection of testosterone propionate (500 microg) during the critical window of rat development (postnatal day 5) induces temporary appearance of aged interstitial cells in developing ovaries (days 7 and 10). Aged interstitial cells showed large size (> or = 12 microm), enhanced androgen receptor (AR) and low estrogen (ER) and luteinizing hormone receptor (LHR) expression. Although normal mature interstitial cells (large size and strong ER and LHR expression) appeared later (day 14), and ovaries of androgenized rats were similar to normal ovaries between days 14 and 35, ovaries of adult androgenized females showed only aged and no mature interstitial cells. Androgenization on day 10 caused the development of aged interstitial cells on day 14, but adult ovaries were normal. Long lasting postnatal estrogenization (estradiol dipropionate for four postnatal weeks) caused in developing and adult ovaries a lack of interstitial cell development beyond the immature state. Immature interstitial cells were characterized by a small size (< or = 7 microm) and a lack of AR, ER and LHR expression. Because the critical window for steroid induced sterility coincides with the termination of immune adaptation, we also investigated distribution of mesenchymal cells (Thy-1 mast cells and pericytes, ED1 monocyte-derived cells, CD8 T cells, and cells expressing OX-62 of dendritic cells) in developing and adult ovaries. Developing ovaries of normal, androgenized and estrogenized females were populated by similar mesenchymal cells, regardless of differences in the state of differentiation of interstitial cells. However, mesenchymal cells in adult ovaries showed distinct behavior. In normal adult ovaries, differentiation of mature interstitial cells was accompanied by differentiation of mesenchymal cells. Aged interstitial cells in ovaries of androgenized rats showed precipitous degeneration of resident mesenchymal cells. Immature interstitial cells in ovaries of estrogenized rats showed a lack of differentiation of resident mesenchymal cells. These observations indicate that an alteration of interstitial cell differentiation during immune adaptation toward the aged phenotype results in precipitous degeneration of resident mesenchymal cells and premature aging of ovaries in adult rats, and alteration toward immature phenotype results in a lack of differentiation of mesenchymal cells and permanent immaturity of ovaries in adult females. PMID- 11856553 TI - The synthesis of spermine analogs of the shark aminosterol squalamine. AB - Aminosterols isolated from the dogfish shark Squalus acanthias are promising therapeutic agents in the treatment of infection and cancer. One of these, MSI 1436, has been shown to possess antimicrobial activity slightly better than squalamine. In this study, a series of analogs of MSI-1436 have been synthesized from stigmasterol. The 7 alpha-hydroxy substituent of MSI-1436 was either omitted or the stereochemistry modified to the 7 beta position. Also, analogs of MSI-1436 with 24-sulfate, 24-amino, and 24-hydroxy substituents were synthesized in order to assess the importance of the side chain functional group on antimicrobial activity. All of the analogs possess significant antimicrobial activity, suggesting that substitution at C7 and C24 of the aminosterols plays a minor role in their antimicrobial potency. PMID- 11856554 TI - Regulation of angiopoietin and Tie-2 receptor expression in non-reproductive tissues by estrogen. AB - Estrogen promotes endothelial cell proliferation and survival in the vasculture of non-reproductive organs. The main mechanisms through which estrogen exerts its effects on endothelial cells remain unknown. Angiopoietins are newly described modulators of endothelial cell survival and they exert their effects through the activation of endothelial cell-specific Tie-2 receptors. In this study, we evaluated whether estrogen modulates the activity and expression of Tie-2 receptors, Ang-1 and its endogenous antagonist; angiopoietins-2 (Ang-2) in non reproductive organs. Using RT-PCR, we found that daily administration of 17-beta estradiol for 8 days in ovariectomized rats results in a significant reduction in tissue Ang-1 mRNA expression. By comparison, estrogen therapy produced a significant increase in Ang-2 mRNA in estrogen-treated rats with heart, kidney and lung Ang-2 mRNA levels reaching 169%, 152% and 224% of those of oil-treated animals, respectively. We also observed that tyrosine phosphorylation of Tie-2 receptors is significantly attenuated in ovariectomized rats treated with 17-beta estradiol. Our results suggest that the effects of estrogen on the vasculature of non-reproductive organs require the inhibition of angiopoietin-1-Tie-2 receptor pathway and that this inhibition is achieved through simultaneous down-regulation of Ang-1 and Tie-2 expression and elevation in Ang-2 expression. PMID- 11856556 TI - Brain imaging of the central executive component of working memory. AB - This review presents neuroimaging studies which have explored the cerebral substrates of the central executive component of the working memory model proposed by Baddeley and Hitch [Working memory (1986); Recent advances in learning and motivation (1974)]. These studies have demonstrated that different executive functions (manipulating and updating of information, dual-task coordination, inhibition and shifting processes) not only recruit various frontal areas, but also depend upon posterior (mainly parietal) regions. Such results are in agreement with the hypothesis that executive functions rely on a distributed cerebral network not restricted to anterior cerebral areas. Moreover, the intervention of similar prefrontal regions in a large number of executive tasks suggests that the central executive functioning must be understood in terms of different interactions between a network of regions rather than in terms of a specific association between one region and one higher-level cognitive process. PMID- 11856555 TI - X-ray and deuterium labeling studies on the abnormal ring cleavages of a 5 beta epoxide precursor of formestane. AB - A new convergent synthesis of the antitumor steroid formestane (4-OHA) 5 has been performed from the easily available epimeric mixture of 5 alpha- and 5 beta androst-3-en-17-one 1a and 1b in order to attempt a yield improvement. A two-step oxidative route followed by base-catalyzed isomerization was applied to the 5 alpha- and 5 beta-epimers 1a and 1b, either as a mixture or separately, leading to the title compound 5. From epimer 1a an efficient process was attained to prepare the desired aromatase inhibitor formestane. Epimer 1b led to the formation of the same compound 5. Additionally, 1b have also been converted in 5 beta-hydroxyandrostane-3,17-dione 12 and androst-4-ene-3,17-dione 13, revealing an unexpected reactivity of the 3 beta,4 beta-epoxy-5 beta-androstan-17-one intermediate 6 formed from 1b during the first oxidative step with performic acid. Cleavage of the epoxide 6 led to the trans-diaxial and the trans diequatorial vic-diols 7 and 8 and to the 1,3-diol 9. The formation of the abnormal products 8 and 9 were investigated through X-ray and deuterium labeling studies. Diol 8 was formed through a trans-diequatorial epoxide ring opening and the 1,3-diol 9 was formed through an intramolecular rearrangement involving a 1,2 hydride shift. All the vic-diols 3, 7 and 8 formed, proved to be good precursors for the synthesis of the target compound 5. PMID- 11856557 TI - Physiological effects of sustained blockade of excitatory synaptic transmission on spontaneously active developing neuronal networks--an inquiry into the reciprocal linkage between intrinsic biorhythms and neuroplasticity in early ontogeny. AB - Spontaneous bioelectric activity (SBA) taking the form of extracellularly recorded spike trains (SBA) has been quantitatively analyzed in organotypic neonatal rat visual cortex explants at different ages in vitro, and the effects investigated of both short- and long-term pharmacological suppression of glutamatergic synaptic transmission. In the presence of APV, a selective NMDA receptor blocker, 1-2- (but not 3-)week-old cultures recovered their previous SBA levels in a matter of hours, although in imitation of the acute effect of the GABAergic inhibitor picrotoxin (PTX), bursts of action potentials were abnormally short and intense. Cultures treated either overnight or chronically for 1-3 weeks with APV, the AMPA/kainate receptor blocker DNQX, or a combination of the two were found to display very different abnormalities in their firing patterns. NMDA receptor blockade for 3 weeks produced the most severe deviations from control SBA, consisting of greatly prolonged and intensified burst firing with a strong tendency to be broken up into trains of shorter spike clusters. This pattern was most closely approximated by acute GABAergic disinhibition in cultures of the same age, but this latter treatment also differed in several respects from the chronic-APV effect. In 2-week-old explants, in contrast, it was the APV+DNQX treated group which showed the most exaggerated spike bursts. Functional maturation of neocortical networks, therefore, may specifically require NMDA receptor activation (not merely a high level of neuronal firing) which initially is driven by endogenous rather than afferent evoked bioelectric activity. Putative cellular mechanisms are discussed in the context of a thorough review of the extensive but scattered literature relating activity-dependent brain development to spontaneous neuronal firing patterns. PMID- 11856558 TI - Non-human primate models for investigating fear and anxiety. AB - Non-human primates exhibit similar physiological and behavioral responses to anxiety-inducing situations as humans and have, in fact, been successfully employed in both conditioned (i.e. conflict paradigms) and ethologically based tests of fear/anxiety (i.e. involuntary isolation, social interaction, human threat, predator confrontation). In the last decade, a renewed and growing interest in non-human primate models has resulted from the use of the small callitrichid species in behavioral pharmacology and neuroscience. This review focuses on the available non-human primate models for investigating fear/anxiety, addressing their advantages, shortcomings, and conceptual framework on which they are based. Lastly, a new ethologically based model to study anxiety and fear induced avoidance in callitrichids--the marmoset predator confrontation test--is discussed. PMID- 11856559 TI - Associative deficit accounts of disrupted latent inhibition and blocking in schizophrenia. AB - Latent inhibition (attenuated responding to a signal due to signal-alone presentations preceding the signal-outcome pairings) and blocking (attenuated responding to Signal B due to Signal A being paired with the outcome prior to pairings of an AB compound with the outcome) are reportedly absent in acute schizophrenics. The common assumption that these phenomena reflect the normal functioning of attention and the observation that rats administered low doses of amphetamine show a similar disruption has resulted in the development of an animal model of attentional dysfunction in acute schizophrenia. Here, we selectively review the experimental and clinical literature concerning latent inhibition and blocking, their disruption in acute schizophrenia, and the current status of this model. We conclude that the construct validity of the model is compromised if latent inhibition and blocking are viewed in attentional terms because experimental data indicate both phenomena can be better understood in associative terms. We favor a framework in which disruption of latent inhibition and blocking in acute schizophrenics is viewed as an inability to compare and express stored representations (i.e. associative performance deficit). This change of perspective does not undermine the potential value of the model, but rather suggests that the nature of its validity should be reconsidered. PMID- 11856560 TI - Orienting of attention in left unilateral neglect. AB - After right posterior brain damage, patients may ignore events occurring on their left, a condition known as unilateral neglect. Although deficits at different levels of impairment may be at work in different patients, the frequency and severity of attentional problems in neglect patients have been repeatedly underlined. Recent advances in the knowledge of the mechanisms of spatial attention in normals may help characterizing these deficits. The present review focuses on studies exploring several aspect of attentional processing in unilateral neglect, with particular reference to the dichotomy between 'exogenous', or stimulus-related, and 'endogenous', or strategy-driven, orienting of attention. A large amount of neuropsychological evidence suggests that a basic mechanism leading to left neglect behavior is an impaired exogenous orienting toward left-sided targets. In contrast, endogenous processes seem to be relatively preserved, if slowed, in left unilateral neglect. Other component deficits, such as a general slowing of the operations of spatial attention, might contribute to neglect behavior. These results are presented and discussed, and their implications for hemispheric specialization in attentional orienting and for the mechanisms of visual consciousness are explored. PMID- 11856561 TI - Neural pathways underlying vocal control. AB - Vocalization is a complex behaviour pattern, consisting of essentially three components: laryngeal activity, respiratory movements and supralaryngeal (articulatory) activity. The motoneurones controlling this behaviour are located in various nuclei in the pons (trigeminal motor nucleus), medulla (facial nucleus, nucl. ambiguus, hypoglossal nucleus) and ventral horn of the spinal cord (cervical, thoracic and lumbar region). Coordination of the different motoneurone pools is carried out by an extensive network comprising the ventrolateral parabrachial area, lateral pontine reticular formation, anterolateral and caudal medullary reticular formation, and the nucl. retroambiguus. This network has a direct access to the phonatory motoneurone pools and receives proprioceptive input from laryngeal, pulmonary and oral mechanoreceptors via the solitary tract nucleus and principal as well as spinal trigeminal nuclei. The motor-coordinating network needs a facilitatory input from the periaqueductal grey of the midbrain and laterally bordering tegmentum in order to be able to produce vocalizations. Voluntary control of vocalization, in contrast to completely innate vocal reactions, such as pain shrieking, needs the intactness of the forebrain. Voluntary control over the initiation and suppression of vocal utterances is carried out by the mediofrontal cortex (including anterior cingulate gyrus and supplementary as well as pre-supplementary motor area). Voluntary control over the acoustic structure of vocalizations is carried out by the motor cortex via pyramidal/corticobulbar as well as extrapyramidal pathways. The most important extrapyramidal pathway seems to be the connection motor cortex-putamen-substantia nigra-parvocellular reticular formation-phonatory motoneurones. The motor cortex depends upon a number of inputs for fulfilling its task. It needs a cerebellar input via the ventrolateral thalamus for allowing a smooth transition between consecutive vocal elements. It needs a proprioceptive input from the phonatory organs via nucl. ventralis posterior medialis thalami, somatosensory cortex and inferior parietal cortex. It needs an input from the ventral premotor and prefrontal cortex, including Broca's area, for motor planning of longer purposeful utterances. And it needs an input from the supplementary and pre supplementary motor area which give rise to the motor commands executed by the motor cortex. PMID- 11856562 TI - Measurement of absolute values of hemoglobin oxygenation in the brain of small rodents by near infrared reflection spectrophotometry. AB - Reflection near infrared spectroscopy (reNIRS) has been proposed as a novel technique for the measurement of absolute values of total hemoglobin (tHb), oxygenated hemoglobin (oxHb), hemoglobin saturation (SO2), and cytochrome aa3 oxidation status (oxCyt aa3) in living tissue. In this study, we evaluated reNIRS during physiological cerebral blood flow conditions in rats (n=6) and during the induction of global cerebral ischemia in gerbils (n=6). ReNIRS parameters were assessed over the exposed cerebral cortex and compared to regional cerebral blood flow (rCBF) data obtained by laser Doppler flowmetry. Under physiological conditions, reNIRS measurements reflected the large intra- and interindividual variability of oxHb and tHb in the brain. The absolute values obtained by reNIRS for tHb (6.3 +/- 1.7 mg/ml), oxHb (3.7 +/- 1.1 mg/ml), and SO2 (61 +/- 5%) matched expected values. In contrast, measurements of oxCyt aa3 were unstable and results unreliable. reNIRS reliably detected cerebral ischemia, verified by a reduction of rCBF to 11% of baseline. tHb dropped to 74 +/- 7% of baseline (P<0.001), reflecting ischemic microvascular vasoconstriction. oxHb and SO2 dropped to expected near-zero values (2 +/- 4 and 3 +/- 5% of baseline, respectively; P<0.001). We conclude that reNIRS provides reliable and reproducible absolute values for brain tissue tHb, oxHb, and SO2 in small rodents. Determination of physiological values requires measurements at multiple locations, while cerebral ischemia is reliably detected by continuous recordings at a single location. PMID- 11856563 TI - Dural substitute for long-term imaging of cortical activity in behaving monkeys and its clinical implications. AB - We present a transparent silicone dural substitute, which we have been using for the last 7 years for imaging cortical dynamics in awake behaving monkeys. This substitute enabled us to record optically for more than a year intrinsic or voltage sensitive dye signals. It is thin and elastic enough to allow microelectrode to pass through without any damage, using full visual control to target the electrode to the desirable recording site. This implant has proved crucial for maintaining the cortex in a good physiological condition and for preserving its optical characteristics that are necessary for optical imaging. We describe the details of the surgical implantation of the silicone dural substitute, the maintenance of the exposed cortex over long periods of time, the cortical reaction to this implant and its possible clinical implications in humans, and the rehabilitation procedure in monkeys. PMID- 11856564 TI - A three-dimensional multi-electrode array for multi-site stimulation and recording in acute brain slices. AB - Several multi-electrode array devices integrating planar metal electrodes were designed in the past 30 years for extracellular stimulation and recording from cultured neuronal cells and organotypic brain slices. However, these devices are not well suited for recordings from acute brain slice preparations due to a dead cell layer at the tissue slice border that appears during the cutting procedure. To overcome this problem, we propose the use of protruding 3D electrodes, i.e. tip-shaped electrodes, allowing tissue penetration in order to get closer to living neurons in the tissue slice. In this paper, we describe the design and fabrication of planar and 3D protruding multi-electrode arrays. The electrical differences between planar and 3D protruding electrode configuration were simulated and verified experimentally. Finally, a comparison between the planar and 3D protruding electrode configuration was realized by stimulation and recording from acute rat hippocampus slices. The results show that larger signal amplitudes in the millivolt range can be obtained with the 3D electrode devices. Spikes corresponding to single cell activity could be monitored in the hippocampus CA3 and CA1 region using 3D electrodes. PMID- 11856565 TI - MRI volume measurements of hypointense objects. A phantom study using stereological methods. AB - Volume estimates made from thick slabs overestimate the volume of a positive contrast particle in a translucent matrix and underestimate its volume if the particle has negative contrast and the matrix is opaque. For T2-weighted MRI high signal objects the bias can be corrected according to simple geometric models. For negative contrast or hypointense objects the magnitude of the bias is unknown and no corrections have yet been put forward. We wanted to determine the bias of MRI measurements of hypointense objects of known diameter surrounded by a positive contrast solution. One, 3 and 5 mm slices with three different contrast concentrations were obtained according to a stereological MRI protocol. Results were compared with the true object size to estimate the bias. A simple correction for the bias could not be developed due to interslice cross-talk. Cross-talk makes an object appear in more MRI slices than corresponding to its physical size and larger than the true diameter. For phantom measurements an empirical correction could be developed, but for measurements of non-ideal objects such as brain structures the validity of the correction would be unpredictable. Besides avoiding or reducing cross-talk, the best way to deal with this bias is to measure the object's maximal contrast from edge to edge, a solution which is not perfect. PMID- 11856566 TI - Comparing the efficacy of two fluorescent retrograde tracers in labeling the motor and sensory neuron populations of the rat sciatic nerve. AB - We compared the efficacy with which the fluorescent tracers Fast Blue (FB) and Diamidino Yellow (DY) retrogradely label neutrons. Trace crystals were applied to the sciatic nerve exclusively (single label) or serially (double label). Unbiased cell counts showed that FB and DY label similar numbers of motoneurons (P=1.00, df 5) or DRG neurons (P=0.95, df 5) when applied exclusively. Plotting of motoneurons revealed a similar pattern of distribution of FB and DY labeled neurons. When the tracers were applied serially, 79% of labeled motoneurons and 77% of labeled DRG neurons were double-labeled irrespective of which tracer was applied first. Equal proportions of the remaining labeled neurons were single labeled with FB or DY. These data show that FB and DY label equal numbers of motor and sensory neurons of the sciatic nerve following exclusive or serial application of tracers. These findings support the use of FB and DY together in serial fluorescent labeling experiments. PMID- 11856567 TI - A quantitative histochemical assay for activities of mitochondrial electron transport chain complexes in mouse spinal cord sections. AB - Mitochondrial dysfunction and degeneration are associated with neurodegenerative disorders. A dysfunctional mitochondrial electron transport chain (ETC) impairs ATP production and accelerates the generation of free radicals. To quantify ETC activity, solution-spectrophotometric assays and histochemical reactions on blue native polyacrylamide gel electrophoresis (BN-PAGE) gels have been used. These methods, however, do not provide information regarding mitochondrial ETC activities associated with specific regions in the central nervous system (CNS). Because neurodegenerative diseases often strike a specific subset of neurons within specific regions in the CNS, reliable methods for quantifying mitochondrial ETC activities in selected CNS regions are needed. We have studied the quantitative range of in situ histochemical assays for ETC complex I, II and IV and determined the optimal conditions for quantification of these ETC complex activities. We also demonstrate that these assays can detect a decrease in mitochondrial ETC activities in the ventral horn of spinal cords isolated from a transgenic mouse model for amyotrophic lateral sclerosis (ALS), a fatal neurodegenerative disease. PMID- 11856568 TI - Development and validation of a language learning model for behavioral and functional-imaging studies. AB - To obtain a more precise understanding of the mechanisms involved in language acquisition in healthy subjects and the re-acquisition of language following brain damage, we developed a word learning model which can be used in behavioral and functional-imaging studies. We generated a set of spoken pseudowords, which were normalized with respect to duration and loudness, and (based on a rating study with 40 students) selected 50 pseudowords, which yielded few associations with existing words and were of neutral emotional valence. The selected pseudowords were paired with object drawings in a pseudo-randomized manner and each subject received a different combination of pairings. These auditory-visual pairs were used to train ten subjects solely on the basis of different frequencies of 'correct' and 'incorrect' pairings. This procedure resembles the acquisition of the first words during childhood. During a five-session training protocol, with one session per day, there was a linear increase of the learning curves, which then reached a plateau. Learning remained stable at 1 and 4 weeks retest intervals. Furthermore, subjects correctly translated the pseudowords into their native language, demonstrating the acquisition of word meanings. These findings indicate that the described associative learning program meets the requirements for a robust and ecologically meaningful model to study the neural mechanisms of language learning and plasticity. PMID- 11856569 TI - Tc99m macroaggregated albumin cisternography for the detection of persistent cerebrospinal fluid (CSF) leak in an animal model. AB - The present study describes a new in vivo animal model that enables the detection of cerebrospinal fluid (CSF) leakage after dural injury. A polyethylene catheter (PE 10) was inserted into the subarachnoid space in the lumbar area by a simple surgical procedure and a radioactive isotope Tc99m Macroaggregated Albumin (Tc99m MAA) was injected into the CSF. In the experimental group, a standardised dural puncture was performed in the cervical area. The accumulation of the isotope in the gauze placed over the dural puncture and viewed by a gamma camera as a spot of concentrated radioactivity, was indicative of CSF leakage. In a second group of animals with intact cervical dura the absence of leakage was presented as a picture of sporadic background radioactivity. To demonstrate the effectiveness of the model in detection of invisible leakage, blood was applied over the cervical dural defect in another group of animals and CSF leakage was assessed by the above mentioned isotope detection method. This in vivo model may be used for evaluation of the sealing properties of various materials under physiological and metabolic processes in living tissue. PMID- 11856570 TI - Recording eye movements with video-oculography and scleral search coils: a direct comparison of two methods. AB - A video-based 2D eye-tracking system (EyeLink version 2.04, SR Research Ltd/SMI) was compared with the scleral search coil technique for its performance on recording the properties of fixations and saccadic eye movements. Fixation positions and saccadic properties (amplitude, duration, and peak velocity) were calculated independently from the data of the two systems that recorded eye positions simultaneously. Fixation positions were well correlated between the video and the coil output with an average discrepancy of <1 degree over a tested range of 40 by 40 degree of visual angle. With respect to the saccade analysis, the values measured by the video system were fitted as a linear function of the values measured by the coil system. Highly correlated linear fits with slopes near one were obtained for all the saccadic parameters. Main sequence relationships (amplitudes-duration and amplitude-peak velocity) were also similar for both systems. A disadvantage of the video method is its low sample rate of 250 Hz. The relatively noisier estimate of all parameters of small saccades could be attributed to this low sampling frequency. PMID- 11856571 TI - A telemetry system to chronically record muscle activity in middle-sized animals. AB - Radio-telemetry enables the long-term recordings of biopotentials that may be obtained in freely moving animals without interference by the experimenter. The purpose of this study was to test a fully implantable device for: (1) its transmission range; (2) the characteristics of the transmitted signals; and (3) its actual application in long-term in vivo registration of EMG. Transmission range was tested by changing the device's position relative to the receiver. Computer simulation of the filtering characteristics provided comparison of original and transmitted signals. Implantation of the device in masticatory muscles, followed by analysis of telemetred signals and determination of activity levels allowed for examination of daily muscle use. The implant's transmission range covered the cage size for middle-sized animals with a minimum of signal dropouts. Transmitted signals were marked by (partial) loss of frequencies beyond 50 Hz, decreased amplitude and slightly delayed timing relative to original waveforms. Analysis of the transmitted EMG revealed that the device can be used for prolonged in vivo EMG registration, detection of peak activity levels, and the examination of general muscle use by the time spent at different levels of activity. PMID- 11856573 TI - Exposure of juvenile guppies to three antiandrogens causes demasculinization and a reduced sperm count in adult males. AB - It has been thoroughly established that the fungicide vinclozolin and the persistent DDT metabolite p,p'-DDE, can function as antiandrogens in mammals in a manner similar to the therapeutic antiandrogen flutamide. In mammals, these chemicals bind the androgen receptor and prevent the transcription of the associated genes causing abnormal sexual development and demasculinization. There are few similar studies in fish and so far it has not been demonstrated that these chemicals have any antiandrogenic effects in this group. In the present study, juvenile guppies (Poecilia reticulata) were fed sublethal doses of vinclozolin, p,p'-DDE or flutamide from birth to adulthood. At sexual maturity, we measured a suite of male sexual characteristics that are known to be under androgen control. All three chemicals caused a reduction in the orange display coloration, inhibited gonopodium development, reduced the sperm count and suppressed courtship behaviour, in a manner consistent with antiandrogen action. Only the gonodosomatic index was unaffected by the treatments. In addition, the three chemicals skewed the sex ratio at adulthood and caused delayed maturation. The altered characters are all considered to be important for male mating success and their impairment indicates that antiandrogens may seriously compromise male reproductive fitness. PMID- 11856572 TI - A monoclonal antibody to tryptophan hydroxylase: applications and identification of the epitope. AB - Recombinant rabbit tryptophan hydroxylase (TPH) was expressed in Escherichia coli and purified from inclusion bodies by preparative sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). A mouse monoclonal antibody and rabbit and sheep polyclonal antibodies were generated. In immunohistochemical studies of formaldehyde-fixed primate brain, the monoclonal strongly labeled not only cell bodies in the raphe nuclei but also fibers in the cerebral cortex. Truncation mutants and peptide pre-competition were used to localize the epitope to E103SVPWFP109. Although the primary sequences of TPH encoded by mRNAs from brain and pineal gland are identical, differences in the immunoreactivity of TPH protein from these two sources were observed in blot immunolabeling studies. TPH immunoreactivity migrated as an M(r) approximately equal 56000 band in each of the tissues except human pineal glands, in which the TPH reactivity was approximately 3 kDa lower. In addition, the relative intensities of TPH immunolabeling across the four tissues differed among these antibodies and a previously described monoclonal antibody against phenylalanine hydroxylase (PH8), which cross-reacts with TPH. Whereas PH8 exhibited roughly equivalent TPH reactivity per protein in both tissues from both species, TPH from human and rat raphe nuclei was preferentially recognized by the present monoclonal. By contrast, the affinity-purified sheep polyclonal antibody reacted preferentially with TPH from human and rat pineal gland, and the affinity-purified rabbit polyclonal antibody appeared to selectively recognize TPH from human pineal gland. PMID- 11856574 TI - Trichloroacetic acid fate and toxicity to the macrophytes Myriophyllum spicatum and Myriophyllum sibiricum under field conditions. AB - Trichloroacetic acid (TCA) has been detected in rain, snow, and river samples throughout the world. It may enter into natural water systems via herbicide use, as a by-product of water disinfection, from emissions of spent bleach liquor of kraft pulp mills, and as a natural fungal product. This compound is phytotoxic and likely to accumulate in aquatic environments. A study to assess the fate of TCA in semi-natural aquatic environments and the toxicity of TCA to rooted aquatic macrophytes was conducted. The experiment involved exposing three replicate 12000 l aquatic microcosms at the University of Guelph Microcosm Facility to 0.05, 0.5, 3, and 10 mg/l of TCA for 35 days in a one-way analysis of variance design. Each microcosm was stocked with 14 individual 5 cm apical shoots of Myriophyllum spicatum and M. sibiricum. The plants were sampled at regular intervals and assessed for the somatic endpoints of plant length, root growth, number of nodes and wet and dry mass and the biochemical endpoints of chlorophyll a and chlorophyll-b, carotenoid content, and citric acid levels. TCA half-lives in the microcosms ranged from 190 to 296 h depending on the initial concentration of TCA. Myriophyllum spp. results indicate that while there were some statistically significant differences from controls, there were no biologically significant effects of TCA for any of the endpoints examined. These data suggest that TCA does not pose a significant risk to these macrophytes up to 10 mg/l, which typically exceeds environmentally relevant concentrations by several orders of magnitude. PMID- 11856575 TI - Community-level disruptions among zooplankton of pond mesocosms treated with a neem (azadirachtin) insecticide. AB - A natural, plant-derived insecticide, neem, is being evaluated as an alternative insect pest control product for forestry in Canada. As part of the process to investigate the environmental safety of neem-based insecticides, a mesocosm experiment was conducted to assess the effects of neem on natural zooplankton communities. Replicate (n=5), shallow (<1 m) forest pond enclosures were treated with Neemix 4.5, at concentrations of 0.035 (the expected environmental concentration), 0.18, 0.70, and 1.75 mg/l active ingredient, azadirachtin. Zooplankton communities were quantitatively sampled over a 4-month experimental period in treated and control enclosures, and water samples were collected to track azadirachtin concentrations. Concentrations in water declined linearly with estimated DT(50) values of 25-29 days. Trends in abundance over time among populations of cladocerans, copepods, and rotifers were found to differ significantly among treatments. At the two highest test concentrations, adverse effects were obvious with significant reductions in several cladoceran species, and near elimination of the three major copepod species present. More subtle effects at the two lowest test concentrations were determined by comparing the community structure of enclosures across treatment levels and over time through an analytical process based on the multivariate statistical software, PRIMER. Significant effects on community structure were detected at both of these lower concentrations, including the expected environmental concentration of 0.035 mg/l azadirachtin. Differential responses among species (some increases, some decreases) caused detectable disruptions in community structure among zooplankton of treated enclosures. Perturbations to zooplankton communities were sufficient to cause measurable differences in system-level metabolism (midday dissolved oxygen concentrations) at all but the lowest test concentration. PMID- 11856576 TI - Glucose-6-phosphate dehydrogenase: the key to sex-related xenobiotic toxicity in hepatocytes of European flounder (Platichthys flesus L.)? AB - The role of glucose-6-phosphate dehydrogenase (G6PDH) in oxidative stress responses was investigated in isolated intact living hepatocytes of immature female and male European flounder (Platichthys flesus L.) because it is the major provider of NADPH needed as reducing power for various detoxification pathways. Hepatocytes were exposed to sublethal concentrations of effective prooxidants such as 100 microM hydrogen peroxide (H(2)O(2)), 100, 200 and 400 microM benzo[a]pyrene (B[a]p) and 50 microM nitrofurantoin (NF) during culture. Since there is evidence that 17-beta-estradiol inhibits certain pathways of xenobiotic biotransformation, we tested also the effects of different concentrations of 17 beta-estradiol (0.2, 1 and 2 microM) alone and 1 microM in combination with 100 microM B[a]p on G6PDH activity. After short-term (1 day) and long-term (9 days) exposure, G6PDH activity was quantified in intact living hepatocytes by a tetrazolium salt method using tetranitroblue tetrazolium salt (TNBT). Hepatocytes obtained from male fish generally showed higher G6PDH activity than those of females. We observed significant inhibition of G6PDH activity by all oxidative stressors and 17-beta-estradiol in both sexes of fish independently of culture conditions, but inhibition was stronger in cells of females than in cells of males. A cumulative effect of the steroid and B[a]p was not found. Our results indicate a sex-dependent inhibitory effect of all stressors and 17-beta-estradiol on G6PDH activity in flounder hepatocytes independent of prooxidant activity of the specific compound. Consequently, NADPH supply for xenobiotic detoxification and other cellular antioxidative defence mechanisms may be different in livers of female and male flounder. The strongly decreased supply of NADPH in hepatocytes of females may explain the reduced and/or delayed NADPH-dependent activity of xenobiotic biotransformation systems such as cytochrome P450 (CYP450) and a lower capacity of non-enzymatic defence systems such as reduced glutathione that is particularly observed in female flounder. Moreover, the strong inhibition of G6PDH in livers of female flounder may explain higher susceptibility for xenobiotic toxicity and, therefore, potentially a higher risk to develop liver cancer. PMID- 11856577 TI - Exposure of Japanese medaka (Oryzias latipes) to benzo[a]pyrene suppresses immune function and host resistance against bacterial challenge. AB - Besides being a potent chemical carcinogen, benzo[a]pyrene (BaP) has also been shown to suppress the immune response of mammals. However, even though BaP is a ubiquitous environmental contaminant to which aquatic species may be directly exposed, information regarding the effects of BaP on the immune system of fish is still lacking. Therefore, laboratory studies were conducted using Japanese medaka (Oryzias latipes) to examine the effects of BaP on host immune status. A single IP injection of BaP at 2, 20 or 200 microg/g BW had no effect upon medaka survival or condition factors for up to 7 days post-injection. Forty-eight hours after injection of either BaP or the vehicle control, fish were sacrificed and the appropriate organs/cells used to assess effects upon: splenic lymphocyte proliferation; kidney phagocyte intracellular superoxide (*O(2)(-)) production; and, CYP1A protein level/activity. In separate experiments, fish were injected with either sheep red blood cells or the bacterial pathogen Yersinia ruckeri at 48 h post-BaP exposure for later determination of antibody-forming cell (AFC) numbers and bacterial host resistance, respectively. Results demonstrated that in the absence of effects upon host survival or condition factors, a single exposure to a relatively low dose of BaP (2 microg/g BW) significantly suppressed mitogen stimulated T- and B-lymphocyte proliferation (in the absence of elevated hepatic CYP1A expression/activity). At higher concentrations, BaP also reduced AFC numbers, phagocyte-mediated *O(2)(-) production, and host resistance against bacterial infection. These results clearly demonstrate the ability of BaP to compromise the immune response of fish and indicate the utility of the fish immune response to serve as an early indicator of BaP exposure/effects in exposed feral populations. PMID- 11856578 TI - Mercury accumulation and flux across the gills and the intestine of the blue crab (Callinectes sapidus). AB - This paper details the results of perfusion experiments examining the accumulation of inorganic and methylmercury (Hg and MMHg) into the gill and intestine tissue of the blue crab, Callinectes sapidus. Additionally, the flux across the tissue to an internal medium, representative of crab tissue or haemolymph, during the perfusion was also measured. The accumulation and transfer processes were studied for each form by exposing the organs to a wide range of Hg and MMHg water concentrations, as well as a mixture of the two Hg forms. Experiments were also performed at different temperatures and in the presence of a metabolic inhibitor to assess the accumulation mechanisms. While the Hg levels bioaccumulated in the two organs were of the same order, the fluxes of Hg from the tissue to the internal medium were slightly higher in the intestine than in the gill. At low external concentrations, the uptake was very similar for both Hg forms, but as exposure pressure increased, inorganic Hg uptake slowed whereas MMHg uptake increased linearly. The results from the perfusion experiments with a mixture of inorganic Hg and MMHg show that while these two forms of Hg do share common uptake pathways, there is also independent uptake. The temperature and inhibition experiments with ouabain, a Na(+)K(+)ATPase inhibitor, show that accumulation is at least partially energy dependent. Overall, the results suggest that there is more than one mechanism of accumulation for both Hg forms. Finally, as accumulation of Hg and MMHg into these tissues was similar, these results contrast with the literature assertion that the enhanced bioaccumulation of MMHg over inorganic Hg is a result of MMHg being more readily transported across the gut membrane. PMID- 11856579 TI - Induction of protective immunity by topic application of a recombinant adenovirus expressing rabies virus glycoprotein. AB - The objective of this study was to determine if a replication defective recombinant adenovirus expressing rabies virus glycoprotein (Adrab.gp) given through a non-invasive vaccination route (by topical application) onto the skin (NIVS) could elicit an immune response and/or protection against rabies. Groups of mice were immunized by NIVS with various doses of Adrab.gp. For comparison, groups of mice were immunized intramuscularly, subcutaneously, or intradermally with Adrab.gp. Mice received two booster immunizations at 1 and 2 months after the first immunization. Virus neutralizing antibody (VNA) titers were measured at day 21 after the first and second immunizations and at day 14 after the third immunization. Fifty percent of the mice immunized by NIVS with 2 x 10(7) and 2 x 10(8)pfu Adrab.gp vaccine developed VNA, whereas none of the control mice or the mice immunized by NIVS with the lowest dose (2 x 10(6)pfu) of Adrab.gp virus developed VNA. However, this low dose induced high titers of VNA in mice immunized by parenteral routes. Two weeks after the last immunization, all the mice were challenged with a lethal dose of rabies virus. More than 70% of the animals immunized by NIVS with > or = 2 x 10(7)pfu Adrab.gp virus survived the challenge, whereas all the mice in the negative control group and the group immunized by NIVS with the lowest dose of Adrab.gp succumbed to rabies. Taken together, the results suggest that NIVS with Adrab.gp can induce VNA production and protection against lethal challenge with rabies virus in mice. PMID- 11856580 TI - Identification and characterization of Marek's disease virus serotype 1 (MDV1) ICP22 gene product: MDV1 ICP22 transactivates the MDV1 ICP27 promoter synergistically with MDV1 ICP4. AB - A previous report [Virus Genes 6 (1992) 365-378] has shown that the US1 gene of Marek's disease virus serotype 1 (MDV1) encodes a homologue of herpes simplex virus type 1 infected cell protein No. 22 (ICP22). In the present study, we expressed and identified a product of the MDV1 US1 gene in chicken embryo fibroblasts (CEFs) with the aid of a recombinant baculovirus expressing a Flag epitope-tagged MDV1 US1 gene, under control of the SRalpha promoter (composed of the enhancer region of the simian virus 40 early promoter and the R region of the human T-cell leukaemia virus type 1 long terminal repeat). In CEF infected with the recombinant baculovirus, MDV1 ICP22 was specifically and efficiently expressed in the presence of n-butyric acid. The apparent M(r) of the expressed protein was 30,000. Reporter gene assays revealed that MDV1 ICP22 by itself transactivated an MDV1 ICP27 promoter/reporter construct weakly but specifically, and furthermore, worked synergistically with MDV1 ICP4 to efficiently up-regulate the MDV1 ICP27 promoter. MDV1 ICP22 may be a regulatory protein that stimulates viral promoters in co-operation with other viral regulatory proteins such as MDV1 ICP4. PMID- 11856582 TI - Quantitative assessment of clinical signs for the detection of classical swine fever outbreaks during an epidemic. AB - The performance of clinical signs as a diagnostic test for the detection of classical swine fever (CSF) outbreaks during the 1997-1998 CSF epidemic in The Netherlands was evaluated by constructing and analysing a receiver operating characteristic (ROC) curve. This curve assesses the discriminating ability of a diagnostic test over a range of test signals. The cut-off values for a defined diagnostic test to detect CSF outbreaks were set by different combinations of clinical signs observed. The area under the ROC curve, which is a quantitative measure of test performance, was significantly (P<0.001) larger than the area under the random ROC curve. This indicates that clinical signs have a significantly higher performance as a diagnostic test for the detection of CSF than for flipping a coin. However, the gain in diagnostic performance compared to a random process is not as much as we would wish it to be. The optimal efficient diagnostic test combined a sensitivity of 72.7% with a specificity of 52.7%, with a combination of the following clinical signs: unsteady gait/ataxia, not eating, not reacting to antibiotic treatment, conjunctivitis, hard faecal pellets. PMID- 11856581 TI - Analysis of the capsid protein gene of a feline-like calicivirus isolated from a dog. AB - The authors report the sequence analysis of the capsid protein-encoding gene (ORF2) of a calicivirus strain recently isolated in Italy from a pup with enteritis. Sequence comparison and phylogenetic analysis revealed that the isolate is highly similar to field and reference feline caliciviruses (FCVs). The isolation of a feline-like calicivirus and the results of recent serological investigations suggest that FCV infection frequently occurs in dogs, but the consequences of this interspecies infection in the canine host are still unknown. PMID- 11856583 TI - An ELISA for antibodies against infectious bronchitis virus using an S1 spike polypeptide. AB - Using the whole infectious bronchitis virus (IBV) for detecting the antibody against IBV by enzyme-linked immunosorbent assay (ELISA) is a routine work in poultry industry. To prepare virus is time consuming and tedious. Furthermore, the whole viral antigen detects all antibodies against the viral structural proteins, including spike (S), nucleocapsid, matrix, and other proteins. Among those, S protein is related to neutralization. Thus, to develop and express protein fragment from S gene and to use the protein as a coating antigen for antibody detection against IBV are the purposes of this experiment. A partial S gene fragment (n.t. 1143-1665) was cloned into pRSET vectors and transformed into competent Escherichia coli (E. coli) BL21 (DE3). A 27.5 kDa fusion protein (S-fg, containing S1-F and partial S2-G antigenic sites) was successfully expressed, affinity-purified and detected specifically with chicken anti-IBV serum by Western blot. The expressed S-fg protein was used as a coating antigen for developing an ELISA (S-fg ELISA) for serum antibody detection in anti-IBV antisera from different IBV serotypes and in field sera. The results show that the S-fg fusion protein is highly cross-reactive among different IBV serotypes, and the S-fg ELISA is found to be a convenient, economical, and efficient method for antibody detection against IBV. PMID- 11856584 TI - Actinobacillus pleuropneumoniae infections in closed swine herds: infection patterns and serological profiles. AB - Many farrow-to-finish herds are endemically infected with Actinobacillus pleuropneumoniae. In order to control the disease efficiently, a better knowledge of the ages at which pigs become infected is necessary. Furthermore, no information is available concerning the influence of maternally derived antibodies on the colonization of the upper respiratory tract. Therefore, A. pleuropneumoniae infection patterns were studied in five farrow-to-finish pig herds (A-E) with a history of pleuropneumonia. A longitudinal study was carried out in herds A and B. In these herds, piglets from sows carrying A. pleuropneumoniae in their noses or tonsils were sampled. Nasal and tonsillar swabs as well as sera, were collected from these animals at the age of 4, 8, 12, 16 (herds A and B) and 23 weeks (herd B). At these ages other pigs from the same sows were euthanized. The lungs were macroscopically examined and samples from nose, tonsils and lungs were collected at necropsy. A cross-sectional study was performed in herds C-E. In these herds nasal and tonsillar swabs, as well as sera, were taken from 10 animals of 4, 8, 12 and 16 weeks of age. Lung, nasal and tonsillar samples were tested for the presence of A. pleuropneumoniae by routine bacteriology and PCR with mixed bacterial cultures. The sera were examined for the presence of Apx toxin neutralizing antibodies. In herd A, A. pleuropneumoniae serotype 2 and 10 strains were isolated, whereas serotype 2, 3, 5b and 8 strains were demonstrated in herd B. In most herds, A. pleuropneumoniae was detected in mixed bacterial cultures of tonsillar and/or nasal samples by PCR from the age of 4 weeks onwards. Colonization of the lungs and development of lung lesions was observed in 12- and 16-week-old animals of herd A and 23-week-old animals of herd B. In most herds, high antibody titres were detected in 4-week-old piglets. These titres decreased during the first 12 weeks of age, but thereafter, increased. It was concluded that PCR with mixed bacterial cultures from tonsillar swabs is a valuable tool for the detection of infected animals. It was also concluded that colonization of tonsils and nasal mucosae can occur in the presence of maternally derived antibodies. Infection of the upper respiratory tract without lung involvement did not result in development of Apx toxin neutralizing antibodies. Therefore, such serological assays cannot be used for the detection of subclinically infected animals. PMID- 11856585 TI - Susceptibility of Arcanobacterium pyogenes from different sources to tetracycline, macrolide and lincosamide antimicrobial agents. AB - Chlortetracycline, oxytetracycline, and the macrolide, tylosin, are extensively used for growth promotion and disease prophylaxis in the cattle and swine industries in the US. Arcanobacterium pyogenes, a common inhabitant of the mucosal surfaces of cattle and swine, is also a pathogen associated with a variety of infections in these animals. A broth microdilution technique was used to determine the antimicrobial susceptibility of 48 A. pyogenes isolates to macrolides, lincosamides and tetracyclines. The MIC50 and MIC90 for chlortetracycline were 0.12 and 8 mg/l, respectively. Similarly, the MIC50 and MIC90 for oxytetracycline were 0.25 and 8 mg/l, while the MIC50 and MIC90 for tetracycline were 0.25 and 16 mg/l, respectively. The MIC50 and the MIC90 were < or = 0.06 and >64 mg/l, respectively, for erythromycin, tylosin and clindamycin. This resistance pattern indicated that some of these A. pyogenes isolates may carry an MLS(B) resistance determinant. A. pyogenes isolates (12.5%) were resistant to erythromycin, and this percentage doubled when MICs were performed following induction with erythromycin. Of the 48 A. pyogenes isolates, 25 and 41.7% were resistant to MLS(B) antimicrobial agents and the tetracycline derivatives, respectively. MLS(B) resistance was present in 22.2 and 35.3% of A. pyogenes isolates of bovine (n=27) or porcine (n=17) origin. In contrast, 70.6% of porcine isolates were resistant to the tetracyclines, compared with 25.9% of bovine isolates. These data suggest that a large proportion of A. pyogenes field isolates may be resistant to these commonly used antimicrobial agents. PMID- 11856586 TI - Haemolytic Escherichia coli isolated from dogs with diarrhea have characteristics of both uropathogenic and necrotoxigenic strains. AB - Twenty-four haemolytic Escherichia coli strains were isolated from dogs with diarrhea. The strains were serotyped and analysed by polymerase chain reaction (PCR) for genes encoding virulence factors associated with E. coli that cause diarrhea in animals. Adhesion antigen production was deduced from haemagglutination experiments. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) of heat extracts was also used as an indication for the production of adhesive structures. The majority of the strains was shown to produce this type of virulence factor. Adhesion and invasion tests of the strains and Caco-2 cells showed that all strains adhered and that two were invasive. The two invasive strains were positive in the intimin PCR and one of them also contained genes encoding CS31A. The PCR for heat stable toxin (ST) was positive in only four strains, as was the presence of F17 fimbrial genes. Surprisingly, 19 strains had intact P fimbrial operons, coding for an adhesin involved in urinary tract infection (UTI). The cytotoxic necrotising factor 1 (CNF1) gene, also mainly found in UTI was likewise detected in these 19 strains. Cytolethal distending toxin (Cdt) genes were found in five strains. The high number of strains positive for CNF1 and P fimbriae prompted us to test the strains in a multiplex PCR used to test E. coli isolated from UTI in various species for 30 virulence associated genes. The data showed that the majority of the diarrhea isolates have virulence factor profiles highly similar to UTI E. coli isolates from dogs. This raises the question whether these isolates are real intestinal pathogens or "innocent bystanders". However, since CNF1 producing necrotoxic E. coli (NTEC) strains isolated from humans, pigs and calves with diarrhea appear to be highly related to our strains, it might be that in dogs this type of isolate is capable of causing not only UTI, but also diarrhea. If this is the case and this type of isolate is "bifunctional", domestic animals likely constitute a reservoir of NTEC strains which can be also pathogenic for humans. PMID- 11856587 TI - Use of recombinant proteins in antibody tests for bovine tuberculosis. AB - Tuberculosis (TB) in cattle remains a major zoonotic and economic problem in many countries. Since the standard diagnostic assay, the intradermal test (IDT) with bovine PPD tuberculin, has less than optimal accuracy in all situations, other diagnostic methods such as serological assays have been investigated. Because of fundamental concerns for the low sensitivity and specificity of previous ELISA protocols, a profiling ELISA with nine purified, recombinant proteins of TB complex mycobacteria, was employed on samples from four groups of cattle: (a) naturally Mycobacterium avium-exposed and experimentally Mycobacterium bovis infected, (b) officially-certified TB-free herds, (c) exposed to M. bovis in two field TB outbreaks and scored as bovine reactors in the gamma-IFN assay for bovine TB, (d) paratuberculosis (para TB)-infected. The described ELISA proved to be highly specific. In fact, the antibody (Ab) response could be consistently detected in 3 out of 3 endotracheally-infected calves and in 1 out of 3 contact infected calves. There was also a very low prevalence of low-titered, non specific Ab responses in paraTB-infected animals. As for the animals exposed to field TB outbreaks, 16 out of 28 gamma-IFN positive cattle were also Ab-positive; importantly, 7 out of 12 gamma-IFN positive, IDT-negative cattle showed Ab responses to TB proteins. In general, the profile of the Ab response varied among animals; the reaction to single recombinant antigens was sometimes transient and fluctuating, whereas the panel of antigens on the whole was indeed more effective in Ab detection. PMID- 11856588 TI - Is there a role of high-dose chemotherapy in lymphoma and solid tumours: the present state of the art. PMID- 11856589 TI - High-dose chemotherapy for solid tumors: results of the EBMT. AB - The European Group for Blood and Marrow Transplantation (EBMT), formerly known as European Group for Bone Marrow Transplantation, was established in 1974 in the Netherlands to share experiences, to promote research and clinical studies and to set up registries in the field of hematopoietic tissue transplantation. At the present time more 400 European and non-European centers are members of the EBMT group. In 1984 a new Working Party was created (Solid Tumors) with the aim to investigate the role of high-dose chemotherapy and stem cell support in the fields of adult and pediatric solid tumors. By January 2000 more than 14000 patients were registered, and at the present time this Registry is the world largest database on this subject. Several phase III randomized clinical trials have recently started on behalf of the Group in different diseases (breast carcinoma, small cell lung cancer, ovarian carcinoma, germ cell tumors and Ewing's family sarcoma). Hundreds of randomized patients will finally produce clearer information on this still experimental therapeutic modality. This paper will describe the EBMT Solid Tumors Working Party Registry updated results as well as the main ongoing studies. PMID- 11856591 TI - The role of high-dose chemotherapy in the treatment of non-small cell lung cancer. AB - The literature on high-dose chemotherapy in non-small cell lung cancer (NSCLC) with autologous bone marrow or peripheral blood stem cell transplantation does not - as of yet - provide evidence of relevant benefits. At the same time, the significant risks of treatment-related morbidity and mortality associated with dose-intensified chemotherapy in this vulnerable patient population are increasingly recognized. Whether the advent of new cytotoxic agents such as the Taxans or newer Topoisomerase inhibitors will help to improve the hitherto unsatisfying results of high-dose chemotherapy in NSCLC, remains to be determined. The few ongoing studies in the area strive to examine such newer drug combinations in a multimodality treatment concept combining neo-adjuvant chemotherapy or chemoradiation with surgery and adjuvant thoracic radiation therapy. PMID- 11856590 TI - High-dose chemotherapy in advanced breast cancer. AB - Based on in vitro and animals studies which assess dose effect relationship specially for alkylating agent, and on the importance on dose intensity in human protocols, high-dose chemotherapy with stem cell support has been widely evaluated in various tumours, particularly in breast cancer. Moreover, in the last few years, the utilization of hematopoietic growth factors and peripheral stem cells has permitted a large diffusion of this approach. However, there is not yet clear data on the place of such a treatment in breast cancer. Few randomized trials are available, with mature data. Only one shows an advantage for high-dose therapy in metastatic disease. In adjuvant setting, sample sizes are too small or follow-up not long enough to draw any definitive conclusion on the place of high-dose consolidation chemotherapy in breast cancer. In inflammatory breast cancer, which is a much more less frequent disease, encouraging results have been published in phase two studies, looking at pathological response, or in pilot studies. The next few years will give a mature date of randomized trials which evaluate high-dose chemotherapy given after conventional treatment in metastatic or high risk disease. Effort should be done to better evaluate this strategy in terms of cost and quality of life and to design new studies aimed to evaluate front line multiple intensification. PMID- 11856592 TI - High-dose chemotherapy in adult soft tissue sarcoma. AB - Soft tissue sarcomas represent a rare and heterogeneous disease. Only few drugs have been identified to be active, with doxorubicin, epirubicin and ifosfamide being the only agents with response rates above 20%. Combination chemotherapy results in higher response rates, however, superiority against single agent chemotherapy in terms of survival has not been established yet. Since a dose response relationship is suggested for the anthracyclines and especially ifosfamide, high-dose or dose-intensive chemotherapy with bone marrow or stem cell support has been evaluated by several investigators. The studies are usually small, and included a very heterogeneous group of patients. Randomized trials have not been done, so that definite conclusions cannot be drawn to date. High dose chemotherapy in soft tissue sarcoma has to be considered highly investigational and should not be performed outside clinical trials. Future studies should be focused on the development of active regimens, resulting in complete remission rates, that can be expected to translate into longer survival. Finally, well designed and appropriately powered randomized trials, using established prognostic and predictive factors, should be carried out, preferably in younger patients and in the context of a potentially curative multimodality approach. PMID- 11856593 TI - High-dose chemoradiotherapy (HDC) in the Ewing family of tumors (EFT). AB - EFT is defined by the expression of ews/ets fusion genes. The type of the fusion transcript impacts on the clinical biology. EFT requires risk adapted treatment. A risk-adapted treatment is determined by tumor localisation, tumor stage and volume. For metastatic and relapsed disease the pattern of spread and the time of relapse are the determinants of risk stratification. Staging of Ewing tumors has been considerably improved by magnetic resonance imaging and modern isotope scanning techniques. However, the determination of the extent of the metastatic spread in particular number of involved bones remains an unresolved issue. The prognosis for high-risk Ewing tumors has been improved by multimodal and high dose radio/chemotherapy (HDC). The concepts for high-dose therapy in Ewing tumors are based on dose response and dose intensity relationships. In single agent HDC most experience exists with Melphalan. Several chemotherapeutic agents have been used in combination HDC with or without TBI such as Adriamycin, BCNU, Busulphan, Carboplatin, Cyclophosphamide, Etoposide, Melphalan, Thiotepa Procarbazin and Vincristine. To date, superiority of any high-dose chemotherapy regimen has not been established. However, the clinical biology, the pattern of spread and the time of relapse determine the prognosis of patient who are eligible for HDC. In particular, patients with multifocal bone or bone marrow metastases have a poorer prognosis than patients with lung metastases. In addition, patients with a relapse within 24 months have a poorer prognosis than patients with a relapse later than 24 months after diagnosis. This review will analyze the results of single- and multi-agent chemotherapy with respect to agent combination, dose and risk stratum of patient population. Future therapeutic modalities for the treatment of EFT might encompass immunotherapeutic and genetic strategies including allogeneic stem cell transplantation. PMID- 11856594 TI - High-dose chemotherapy in soft tissue sarcoma in children. AB - Soft tissue sarcomas (STS) are highly malignant tumours that constitute 5-6% of all malignant childhood neoplasms. Of these, rhabdomyosarcoma (RMS) is the most common in children, and has a characteristic two-peak age incidence, 2-5 and 15 19 years. Most children with RMS are cured with conventional chemotherapy and local therapy (surgery with or without radiotherapy). Children with metastatic disease at presentation, particularly those older than 10 years or with bone marrow or bone involvement have a much poorer outcome. In this subgroup, high dose therapy with stem cell rescue has been studied over the last two decades. Various single or multiagent chemotherapy regimens with or without radiotherapy and autologous stem cell rescue have been used as consolidation treatment with little success. Recent trials using sequential high-dose chemotherapy in the early phase of treatment have proved to be feasible, but the beneficial effect has to be confirmed. The role of purging remains unclear. Collaboration between different international groups is urgently required, in an attempt to improve the poor outcome of children with high risk STS. PMID- 11856595 TI - High-dose chemotherapy with autologous stem cell rescue for brain tumors. AB - The use of high-dose chemotherapy with stem cell rescue for malignant brain tumors is reviewed. Promising results have been reported in patients with medulloblastomas, supratentorial PNET's and high-grade astrocytomas. Results thus far have been disappointing for ependymomas and brain stem gliomas. The role of this treatment strategy for other chemotherapy-sensitive tumors such as oligodendrogliomas and central nervous system germ cell tumors has yet to be determined. PMID- 11856596 TI - Prognostic stratification in UPC: a role for assessing the value of conventional dose and high-dose chemotherapy for unknown primary carcinoma. AB - High-dose chemotherapy has been advocated by some investigators as a means to circumvent drug resistance, thereby improving treatment results in patients with solid tumors. For patients with unknown primary tumors, this hypothesis has only recently undergone limited testing. Two groups (one from the USA and one from Europe) have published their experience with higher doses of chemotherapy in the treatment of UPC. The results are not superior to those reported by other investigators using more standard doses of chemotherapy. Most importantly, chemotherapy trials for UPC are usually conducted in small populations made up of heterogeneous patient subsets with varying sensitivity to chemotherapy. It seems likely that progress in the management of patients with unknown primary cancers will occur as a result of efforts to improve the understanding of the natural history of this disease coupled with the assessment of novel agents targeted against specific biochemical abnormalities that will be demonstrated to be important in the development and maintenance of these malignancies. PMID- 11856597 TI - High-dose chemotherapy and autologous bone marrow transplantation in diffuse intermediate- and high-grade non-Hodgkin lymphoma. AB - This is a review of data published in the literature on the role of marrow ablative treatment and blood or marrow transplantation for high-grade and intermediate-grade lymphoma. Timing and epidemiology are reviewed before the various clinical situations (primary refractory partial responses, complete responses and relapses). The Lyon consensus conference held in 1998 has also extensively been used and quoted. PMID- 11856598 TI - High-dose therapy for indolent lymphoma. AB - Stem-cell transplantation (SCT) has become the treatment of choice for patients with relapsed aggressive non-Hodgkin's lymphoma (NHL). However, the role of SCT in the management of patients with indolent NHL remains controversial. Indolent follicular lymphomas are diseases which are generally incurable with conventional therapy. Although patients can survive for prolonged periods, the median duration of first remission is approximately 3 years, and subsequent remissions are progressively shorter with time. Emerging evidence suggests that high-dose chemotherapy with SCT leads to prolonged disease-free and overall survival in a subset of patients with indolent NHL. However, there is increasing concern regarding the toxicity of SCT, especially the long-term risk of developing myelodysplastic syndrome. It is still unclear as to when this approach should be used. Poorer outcomes have been obtained in heavily pretreated patients but encouraging results are being reported for patients undergoing SCT early during the course of their disease. Investigators are now focusing on how to improve SCT efficacy in order to eradicate minimal residual disease. Many ongoing studies are especially exploring the impact of stem-cell purging and novel ablative regimens combined with allogeneic transplantation. PMID- 11856599 TI - Tumor cell contamination in re-infused stem cell autografts: does it have clinical significance? AB - Tumor cells frequently contaminate autologous stem cell products in patients having a variety of malignancies. Mobilized peripheral blood stem cells may be less contaminated with tumor cells than bone marrow harvests are, but they are still frequently infiltrated. Gene-marking studies using retroviral vectors provide evidence that tumor cells contained in autografts contribute to relapse in myeloid leukemia and neuroblastoma patients. Also clinical studies have shown that tumor cell contamination of autografts is associated with shortened disease free survival; on the other hand, successful ex vivo purging of tumor cells is associated with superior clinical outcome. However, the presence of tumor cells in autografts or insufficient purging may correlate with the extent of systemic residual disease and/or tumor chemosensitivity; therefore, there is no direct evidence that reinfused tumor cells alone cause relapse. Particularly in patients having highly chemosensitive disease and no detected systemic residual disease following high-dose transplant chemotherapy, the relative number of tumor cells contained in autografts and eventually reinfused, may become a determining factor for clinical outcome. There are no randomized trials showing improved (disease free) survival with purging. In the absence of such trials, the contribution of tumor cells in the stem cell autografts to subsequent relapse remains controversial. PMID- 11856600 TI - Economic aspects of high-dose chemotherapy: a clinician's perspective review. AB - Over the last 15 years, the increased use of high-dose chemotherapy (HDC) has led to a considerable increase in the cost of cancer treatments. After making a general economic analysis of the benefits and costs of healthcare initiatives, this paper considers all of the different phases and elements of HDC, as well as the strategies for reducing basic, indirect and out-of-pocket costs. The cost of HDC has decreased by 40-60% over the last decade and its cost-effectiveness ratios are now similar or only slightly higher than those of other widely accepted medical interventions. However, except in the case of some hematological and paediatric neoplasms, the efficacy of the treatment has not yet been clearly defined and so it should only be used in well-designed clinical trials that should also include prospective cost evaluation measures. PMID- 11856602 TI - Applications of single nucleotide polymorphisms in crop genetics. AB - The discovery of single nucleotide polymorphisms (SNPs) and insertions/deletions, which are the basis of most differences between alleles, has been simplified by recent developments in sequencing technology. SNP discovery in many crop species, such as corn and soybean, is relatively straightforward because of their high level of intraspecific nucleotide diversity, and the availability of many gene and expressed sequence tag (EST) sequences. For these species, direct readout of SNP haplotypes is possible. Haplotype-based analysis is more informative than analysis based on individual SNPs, and has more power in analyzing association with phenotypes. The elite germplasm of some crops may have been subjected to bottlenecks relatively recently, increasing the amount of linkage disequilibrium (LD) present and facilitating the association of SNP haplotypes at candidate gene loci with phenotypes. Whole-genome scans may help identify genome regions that are associated with interesting phenotypes if sufficient LD is present. Technological improvements make the use of SNP and indel markers attractive for high-throughput use in marker-assisted breeding, EST mapping and the integration of genetic and physical maps. PMID- 11856603 TI - Epialleles - a source of random variation in times of stress. AB - With the advent of biotechnology, epigenetics has gained in respectability. Recently, focus has moved away from the problems caused by the epigenetic silencing of transgenes to the adaptive advantages offered by stochastic epigenetic variation. Epialleles can form in response to environmental and genomic stresses, including polyploidization. They may be important in acclimation to a range of environmental conditions and in stabilizing polyploid genomes. PMID- 11856604 TI - Plant molecular diversity and applications to genomics. AB - Surveys of nucleotide diversity are beginning to show how genomes have been shaped by evolution. Nucleotide diversity is also being used to discover the function of genes through the mapping of quantitative trait loci (QTL) in structured populations, the positional cloning of strong QTL, and association mapping. PMID- 11856605 TI - Tomato mutants as tools for functional genomics. AB - Tomato mutants have been used in genetic studies and breeding for decades, yet only a few tomato mutants have been characterized at the molecular level. Similarly, a wealth of sequence information for tomato is now available but the functions of only a few genes are known. New developments - such as the use of saturated mutant populations, new methods for the detection of mutants and new sequence data - are bridging the gap between tomato genes and their functions. PMID- 11856606 TI - Sorghum bicolor - an important species for comparative grass genomics and a source of beneficial genes for agriculture. AB - A high-resolution genetic, physical, and cytological map of the sorghum genome is being assembled using AFLP DNA marker technology, six-dimensional pooling of BAC libraries, cDNA mapping technology, and cytogenetic analysis. Recent advances in sorghum comparative genomics and gene-transfer technology are accelerating the discovery and utilization of valuable sorghum genes and alleles. PMID- 11856607 TI - Sugarcane genomics: depicting the complex genome of an important tropical crop. AB - In the past few years, approaches such as molecular cytogenetics and the use of molecular markers have permitted significant advances in the establishment of the evolutionary origin and genome structure of sugarcane, an important polyploid crop. The availability of new resources, such as a bacterial artificial chromosome library and a huge collection of expressed sequence tags, has opened the gateway to promising functional analyses on a genomic scale. PMID- 11856608 TI - Comparing Arabidopsis to other flowering plants. AB - Comparisons of the Arabidopsis genomic sequence with sequences from other flowering plants have revealed that substantial colinearity exists between species in the arrangement of genes within chromosomal blocks. Although seen most clearly in short sequences (at the Megabase scale), this colinearity can also be found using dense genetic maps that are based on expressed sequence tags. The genomes of most diploid Angiosperms show evidence of polyploid ancestry, and the resulting duplicated blocks, which have been subject to deletion and rearrangements during evolution, form complex networks of homology both within and between species. These homologies should prove to be of value in exploiting the Arabidopsis sequence to identify candidate genes in defined chromosomal regions within genomes that are less well characterised. PMID- 11856610 TI - Applied plant genomics: the secret is integration. AB - Although concerted efforts to understand selected botanical models have been made, the resulting basic knowledge varies in its applicability to other diverse species including the major crops. Recent advances in high-throughput genomics are offering new avenues through which to exploit model systems for the study of botanical diversity, providing prospects for crop improvement. In particular, whole-genome sequencing has provided opportunities for the broader application of reverse genetics, expression profiling, and molecular mapping in diverse species. PMID- 11856611 TI - Application of gene silencing in plants. AB - Recent studies of gene silencing in plants have revealed two RNA-mediated epigenetic processes, RNA-directed RNA degradation and RNA-directed DNA methylation. These natural processes have provided new avenues for developing high-efficiency, high-throughput technology for gene suppression in plants. PMID- 11856612 TI - The genetics and molecular genetics of terpene and sterol origami. AB - Terpenes and sterols are complex molecules synthesized by equally complex biosynthetic pathways. Recent progress in using the tools of genetics, molecular genetics and genetic engineering to dissect triterpene metabolism in the cytosol, and terpene metabolism in the plastids, has opened up new strategies and avenues of investigation. Most importantly, these studies have enhanced our appreciation of the biological significance of these compounds for plants, and have revealed new secrets of this intriguing biochemistry for practical applications in agriculture and medicine. PMID- 11856613 TI - Limiting nutrients: an old problem with new solutions? AB - Iron and phosphorus are essential minerals for both humans and plants. Advances in our understanding of the molecular mechanisms involved in the mobilization, transport and storage of these minerals now allow us to engineer plants to improve the yield and mineral nutrition of crops. Strategies range from increasing the expression of endogenous genes, such as that encoding the iron storage protein ferritin, to expressing a phytase gene from the fungus Aspergillus in Arabidopsis, thereby allowing the plants to obtain a previously unusable pool of phosphorus. PMID- 11856614 TI - Engineering the plastid genome of higher plants. AB - The plastid genome of higher plants is an attractive target for engineering because it provides readily obtainable high protein levels, the feasibility of expressing multiple proteins from polycistronic mRNAs and gene containment through the lack of pollen transmission. A chloroplast-based expression system that is suitable for the commercial production of recombinant proteins in tobacco leaves has been developed recently. This expression system includes vectors, expression cassettes and site-specific recombinases for the selective elimination of marker genes. Progress in expressing proteins that are biomedically relevant, in engineering metabolic pathways, and in manipulating photosynthesis and agronomic traits is discussed, as are the problems of implementing the technology in crops. PMID- 11856615 TI - How can we deliver the large plant genomes? Strategies and perspectives. AB - The first sequenced plant genome, from the small mustard plant Arabidopsis thaliana, was published at the end of 2000. The sequencing of the rice genome is well under way. The sizes of plant genomes vary by a factor of up to 1000, and many important crop plants have genomes that are several times larger than the human genome. To gain insight into the gene toolbox of plant species, numerous large-scale EST sequencing projects have been launched successfully, and analysis procedures are constantly being refined to add maximum value to the sequence data. In addition, an alternative approach to exclude repetitive noncoding DNA and to enrich sequence libraries for gene-containing genomic regions has been developed. This strategy has the potential to deliver information about both genes and regulatory regions outside the transcribed regions. PMID- 11856616 TI - Human caliciviruses in Europe. AB - Caliciviruses are single-stranded RNA viruses, which are divided into four genera based on their morphology and genomic structure. Viruses from two genera, the Norwalk like viruses and Sapporo like viruses, are a common cause of acute, nonbacterial gastroenteritis in humans. Although the first human calicivirus discovered nearly 30 years ago, much of the epidemiological and biological character of these viruses is only now beginning to unfold. Investigation has been difficult due to a number of factors, the viruses cannot be amplified by in vitro cell culture or animal models and electron microscopy (EM) is often not sensitive enough to detect the viruses in stool samples. Recent advances in molecular diagnostic techniques and the advent of a baculovirus expression system have highlighted the clinical and public health importance of calicivirus in all age groups, their ability to cause infection via a number of transmission routes as well as their considerable genetic diversity. These characteristics, in conjunction with the inability of humans to develop long-term immunity make HuCV an important public health issue in Europe and worldwide. PMID- 11856617 TI - Two-round rapid-cycle RT-PCR in single closed capillaries increases the sensitivity of HCV RNA detection and avoids amplicon carry-over. AB - BACKGROUND: For the detection of hepatitis C virus (HCV) specific nucleic acids the polymerase chain reaction (PCR) is widely used. Rapid-cycle PCR is performed in glass capillaries with the LightCycler instrument and allows PCR including product analysis to be performed within a closed system in about 1 h. Thus, rapid cycle PCR appears especially suitable for routine diagnostic applications. However, the volume of the PCR vessel is restricted to about 20 microl, which may limit the sensitivity of the PCR. To increase its sensitivity two-round or nested primer PCR protocols have been developed. In rapid-cycle PCR first-round PCR products are usually collected from the capillaries by centrifugation, a procedure prone to cross-contamination. OBJECTIVES: Development of a two-round rapid-cycle reverse transcription-polymerase chain reaction (RT-PCR) in single closed LightCycler capillaries for the sensitive detection of HCV RNA in serum or plasma. STUDY DESIGN: A set of two pairs of nested primers was selected. The first-round RT-PCR reaction mixture was separated from the second-round PCR mixture by silicone oil. Reverse transcription followed by the first-round PCR was performed. Then, the second-round mixture was combined with first-round products by a centrifugation step followed by second round PCR during which fluorescence intensities were recorded and used for quantification. RESULTS: To establish the sensitivity of this novel assay a serial dilution of HCV reference standard was used. In plasma samples about 100 IU/ml HCV were consistently detected using the high pure viral RNA kit for nucleic acid purification. This detection limit was found to be about 20 fold increased compared with single round RT-PCR and corresponded to 3.4 IU of HCV per capillary. Using a panel of HCV genotype standards the novel assay exhibited similar sensitivity for all HCV genotypes. The applicability for clinical routine testing was demonstrated by examining 156 clinical samples. CONCLUSION: Two-round RT-PCR with the LightCycler instrument using a single closed capillary throughout the procedure was found ideally suited for rapid (100 min), accurate and sensitive molecular diagnosis of active HCV infections. Since the capillaries remained closed during the procedure carry-over contamination was precluded. PMID- 11856618 TI - Development of hepatitis B virus resistance for lamivudine in chronic hepatitis B patients co-infected with the human immunodeficiency virus in a Dutch cohort. AB - INTRODUCTION: With the introduction of HAART, the HIV-1 has turned from a lethal into a chronic infection in the majority of patients. In homosexual populations, 20% of HIV-1 infected patients suffer from a chronic HBV infection, which may eventually lead to complications of the liver disease because of prolonged survival. Lamivudine is effective in reducing both HIV-1 and HBV viral replication. However, resistance for lamivudine may complicate the course of the HBV disease in HIV-1-infected patients. We, therefore, conducted a retrospective study in HIV-1-HBV co-infected patients on lamivudine therapy. PATIENTS AND METHODS: All HIV-1-HBV co-infected patients who were treated with lamivudine for over 6 months in five major referral clinics in The Netherlands with HBV DNA above 2.0 x 10(5) geq ml(-1) at baseline, were evaluated. Retrospectively, the course of HBV DNA in available serum samples was established. If HBV DNA was detectable with the sensitive PCR-assay, YMDD-analyses of the polymerase gene of the hepatitis B virus was executed with the INNO-LiPA-DR-strip. RESULTS: Forty six patients were evaluated. The median level of HBV DNA at start of lamivudine therapy was 1.31 x 10(9) geq ml(-1) (range 3.5 x 10(5) - 2.0 x 10(10), n=43). Of three patients no baseline sample was available, but since HBV DNA was still above 2.0 x 10(5) geq ml(-1) at week 3, 7 and 11, these patients were included. Median duration of lamivudine therapy was 97 weeks (range 27-263). The percentage of detected mutations was 25 and 52% at 1 and 2 years, respectively. Twenty-two patients ultimately developed a mutation. Both baseline Body Mass Index (BMI) and the decrease in CD4 cell count as a time dependent factor were significantly related to the emergence of mutations. In 10 out of 12 evaluated patients, HBV DNA levels returned to baseline level or even above baseline level after the development of mutant virus. One patient (5%) developed a flare of serum transaminases (ALT>10 x ULN) 24 weeks after first detection of variant virus. CONCLUSION: There is a linear time-dependent appearance of HBV mutations for lamivudine in our population. In a minority of patients (5%), development of a mutation was followed by a significant elevation of serum transaminases. A decline in CD4 cell count, which may indicate less response to HAART, induces a faster emergence of mutations and close surveillance of HBV co-infected patients on therapy may be indicated due to the prolonged survival of HIV-1 patients. PMID- 11856619 TI - Detection of HSV-2 in genital ulcers from STD patients in Dar es Salaam, Tanzania. AB - BACKGROUND: Genital ulcer disease (GUD) is common in many developing countries. Several reports indicate that there is an association with HIV infection. Analysis by polymerase chain reaction (PCR) has demonstrated that the ulcers are frequently caused by herpes simplex type 2 (HSV-2), although HSV-1 is becoming increasingly important in many parts of the world. Comparable studies have not been performed in Tanzania. OBJECTIVES: To determine the prevalence of HSV-2 and HSV-1 in genital ulcers in Dar es Salaam, Tanzania and determine their possible association with HIV infection. STUDY DESIGN: Samples were collected from 70 consecutive patients with GUD attending a clinic for sexually transmitted diseases. Specimens from ulcers were analysed by PCR for the presence of HSV-2 and HSV-1, and sera were examined for antibodies against HSV-2 and HIV. RESULTS AND DISCUSSION: HSV-2 DNA was detected in 64% of the specimens from ulcers while HSV-1 DNA was not found in any of them. Antibodies to HSV-2 and HIV were detected in 79.7 and 42% of the patients' sera, respectively. Although there was a significant positive association between HIV and HSV-2 seropositivity, HSV-2 DNA in genital ulcers was not more prevalent among HIV seropositive than among HIV seronegative individuals. CONCLUSION: The prevalence of HSV-2 antibodies among Tanzanian patients with genital ulcers is very high, and HSV-2 is detected in most of the ulcers. There is an association between infections with HIV and HSV 2, but the relationship is not clear. PMID- 11856620 TI - The humoral response to live and inactivated influenza vaccines administered alone and in combination to young adults and elderly. AB - BACKGROUND: Vaccination is the most effective way for prevention of severe influenza infection, but the present vaccines are not very efficient in the elderly. OBJECTIVE: In this study, we investigated the Ig isotype response to a cold adapted, live, attenuated influenza vaccine (LIV) and a trivalent, subunit, inactivated influenza vaccine (IIV) and the combination of both (LIV/IIV) in young and elderly men, not previously vaccinated against influenza. STUDY DESIGN: LIV and IIV containing the strains for 1998-1999 were used. Forty-seven 19-35 years old and forty-three 58-91 years old were divided in four groups receiving either LIV, IIV, both or placebo. All were bled before and at 4 weeks after vaccination. The hemagglutination inhibition (HAI) antibody response to homologous strains and the enzyme-linked immunosorbent assays (ELISA) IgA, IgG and IgG-subclass responses to recombinant HA proteins representing influenza A and B strains from 90 to 91, and purified virus from an A/Sydney/05/97-like strain were measured. RESULTS: Protective HAI titers of >40 were more frequent (P<0.05) after IIV alone or LIV/IIV than after LIV alone; there was no difference between the young and elderly persons. The highest frequency of titer rises in ELISA was found against the H1N1 recombinant antigen. In young adults, IgG titer rises were more frequent than in elderly given LIV than IIV alone, but not when given the combination. The influenza-specific IgG subclass response after LIV consisted of IgG1 and IgG4 only in the young; after IIV it stimulated of IgG1 and IgG3 in the young and IgG1 alone in the elderly. After vaccination with the LIV/IIV combination IgG1, IgG3 and IgG4 were found in the young and IgG1, IgG3 in the elderly. Among the elderly, the IIV/LIV combination induced the same response rate of specific IgG and IgG1 as in young adults. CONCLUSION: The study illustrates the possibility to correct the age dependent weakening of the immune response to influenza vaccine with a combination of LIV and IIV. PMID- 11856621 TI - Virus isolation as one of the diagnostic methods for dengue virus infection. AB - Virus isolation is the most reliable evidence of infection. In the present study, we isolated virus from serum samples collected from confirmed dengue cases. When data were analyzed based on disease days, dengue viruses were isolated from 28 of 32 serum samples collected on disease day 5 or earlier. When analyzed based on fever days, dengue viruses were isolated from all the serum samples collected on fever day -3 or earlier, and from 10 of 13 samples collected on fever days -2 and -1. Viruses were isolated from one each of the serum samples collected on fever days 0, 1, 2 and 3, respectively. Virus was not, however, isolated from those collected on fever day 4 or later. The results of virus isolation and reverse transcriptase-polymerase chain reaction were consistent in 78 of 82 serum samples. These results suggest that virus isolation is a useful and sensitive technique for confirmation of dengue virus infection, especially when serum samples are collected before fever subsides. PMID- 11856623 TI - Ipsi- and contralateral EEG reactions to transcranial magnetic stimulation. AB - OBJECTIVES: Transcranial magnetic stimulation (TMS) and high-resolution electroencephalography (EEG) were used to study the spreading of cortical activation in 6 healthy volunteers. METHODS: Five locations in the left sensorimotor cortex (within 3cm(2)) were stimulated magnetically, while EEG was recorded with 60 scalp electrodes. A frameless stereotactic method was applied to determine the anatomic locus of stimulation and to superimpose the results on magnetic resonance images. Scalp potential and cortical current-density distributions were derived from averaged electroencephalographic (EEG) data. RESULTS: The maxima of the ipsilateral activation were detected at the gyrus precentralis, gyrus supramarginalis, and lobulus parietalis superior, depending on the subject. Activation over the contralateral cortex was observed in all subjects, appearing at 22plus minus2ms (range 17--28); the maxima were located at the gyrus precentralis, gyrus frontalis superior, and the lobulus parietalis inferior. Contralateral EEG waveforms showed consistent changes when different sites were stimulated: stimulation of the two most medial points evoked the smallest responses fronto-parietally. CONCLUSIONS: With the combination of TMS, EEG, and magnetic resonance imaging, an adequate spatiotemporal resolution may be achieved for tracing the intra- and interhemispheric spread of activation in the cortex caused by a magnetic pulse. PMID- 11856624 TI - Neurophysiological and behavioral concomitants of mild brain injury in collegiate athletes. AB - OBJECTIVES: There is still limited understanding regarding the effect of mild brain injury (MBI) on normal functioning of the human brain with respect to motor control and coordination. To our knowledge, no research exists on how both the accuracy of force production and underlying neurophysiological concomitants are interactively affected by MBI. The aim of this study is to provide empirical evidence that there are at least transient functional changes in the brain associated with motor control and coordination in collegiate athletes suffering from MBI as reflected in alterations of force trajectory patterns and electroencephalogram (EEG) potentials both in time and frequency domains. METHODS: Comparisons of the performance and concomitant EEG waveforms both in time and frequency domains of 6 collegiate athletes with MBI and 6 normal subjects in a series of isometric force production tasks were made. The traditional averaging techniques to obtain the slow-wave movement-related potentials (MRP) and Morlet wavelet transform to obtain EEG time-frequency (TF) profiles associated with task performance were used. Subjects performed isometric force production tasks when the level of nominal force was experimentally manipulated. EEG recordings from the frontal-central areas were analyzed with respect to the accuracy of force production during the ramp phase. RESULTS: Behaviorally, the accuracy of force trajectory performance was considerably impaired in MBI subjects even when the amount of task force was only increased from 25 to 50% maximum voluntary contraction (MVC) within a given subject. Electro-cortically, impaired performance in MBI subjects was associated with alterations in EEG waveforms, amplitude of MRP and TF profiles of EEG. CONCLUSIONS: Both behavioral and electro-cortical data of control subjects generally were comparable with those from subjects with MBI when small amounts of force were regulated. However, differences become apparent as the amount of task force production was increased. Overall our findings identify the presence of transient functional changes in the brain associated with motor control and coordination in subjects suffering from MBI. PMID- 11856625 TI - EEG differences between good and poor responders to methylphenidate and dexamphetamine in children with attention-deficit/hyperactivity disorder. AB - OBJECTIVES: This series of studies investigated (1) electroencephalographic (EEG) differences between good and poor responders to methylphenidate, (2) EEG differences between good and poor responders to dexamphetamine, and (3) differences in the EEGs of good responders to methylphenidate versus dexamphetamine, within samples of children with the combined type of attention deficit/hyperactivity disorder (ADHD). METHODS: Twenty good and 20 poor responders to each of methylphenidate and dexamphetamine, based on the results of a continuous performance task, and 20 age-matched control subjects, participated in this study. EEG was recorded from 21 sites during an eyes-closed resting condition and Fourier transformed to provide estimates for total power, and absolute and relative power in the delta, theta, alpha and beta bands, and for the theta/alpha and theta/beta ratios. RESULTS: EEG differences were found between the good and poor responders to each medication. Good responders to methylphenidate had EEG profiles that suggested that they were more cortically hypoaroused than poor responders. In contrast, the good responders to dexamphetamine appeared to be more maturationally lagged than the poor responders. The two good-responder groups had EEG profiles which suggested that there were two different underlying central nervous system (CNS) dysfunctions. CONCLUSIONS: Children with the combined type of ADHD do not constitute a homogeneous clinical group, as different types of CNS dysfunction are present within this population. These results also indicate the need for medication testing to be undertaken before a child is prescribed stimulant medication for ADHD. PMID- 11856626 TI - Trial-to-trial variability of cortical evoked responses: implications for the analysis of functional connectivity. AB - OBJECTIVES: The time series of single trial cortical evoked potentials typically have a random appearance, and their trial-to-trial variability is commonly explained by a model in which random ongoing background noise activity is linearly combined with a stereotyped evoked response. In this paper, we demonstrate that more realistic models, incorporating amplitude and latency variability of the evoked response itself, can explain statistical properties of cortical potentials that have often been attributed to stimulus-related changes in functional connectivity or other intrinsic neural parameters. METHODS: Implications of trial-to-trial evoked potential variability for variance, power spectrum, and interdependence measures like cross-correlation and spectral coherence, are first derived analytically. These implications are then illustrated using model simulations and verified experimentally by the analysis of intracortical local field potentials recorded from monkeys performing a visual pattern discrimination task. To further investigate the effects of trial-to-trial variability on the aforementioned statistical measures, a Bayesian inference technique is used to separate single-trial evoked responses from the ongoing background activity. RESULTS: We show that, when the average event-related potential (AERP) is subtracted from single-trial local field potential time series, a stimulus phase-locked component remains in the residual time series, in stark contrast to the assumption of the common model that no such phase-locked component should exist. Two main consequences of this observation are demonstrated for statistical measures that are computed on the residual time series. First, even though the AERP has been subtracted, the power spectral density, computed as a function of time with a short sliding window, can nonetheless show signs of modulation by the AERP waveform. Second, if the residual time series of two channels co-vary, then their cross-correlation and spectral coherence time functions can also be modulated according to the shape of the AERP waveform. Bayesian estimation of single-trial evoked responses provides further proof that these time-dependent statistical changes are due to remnants of the evoked phase-locked component in the residual time series. CONCLUSIONS: Because trial-to-trial variability of the evoked response is commonly ignored as a contributing factor in evoked potential studies, stimulus-related modulations of power spectral density, cross-correlation, and spectral coherence measures is often attributed to dynamic changes of the connectivity within and among neural populations. This work demonstrates that trial-to-trial variability of the evoked response must be considered as a possible explanation of such modulation. PMID- 11856627 TI - High-resolution EEG: on the cortical equivalent dipole layer imaging. AB - BACKGROUND: Brain electrical activity is a spatio-temporally distributed process. Cortical imaging techniques have been developed to reconstruct cortical activity from the scalp electroencephalographic or magnetoencephalographic measurements. Several cortical imaging approaches, such as the epicortical potentials and a dipole layer accounting for the cortical activity, have been used to represent brain electrical activity. METHODS: A closed cortical dipole layer source model is used to equivalently represent brain electrical activity. The relationship between the primary brain electrical sources and the cortical equivalent dipole layer is derived from the theory of electromagnetics. Computer simulation studies were conducted using a 3-concentric-sphere head model to validate the proposed theory. The cortical equivalent dipole layer imaging approach was tested in both computer simulation and human visual evoked potential (VEP) experiments. RESULTS: The strength of the cortical equivalent dipole layer is shown to be proportional to the electrical potential over the same surface generated by primary electrical sources, had the outer medium been replaced by air. The proposed theory was validated by computer simulation in a discrete system. Simulation and VEP experimental studies suggest the feasibility of applying the cortical equivalent dipole layer imaging approach for brain imaging. CONCLUSIONS: The cortical equivalent dipole layer model can equivalently represent the primary brain electrical sources throughout the entire brain surrounded by the dipole layer. The strength of the cortical equivalent dipole layer due to primary sources can be directly calculated according to the theory developed in the present study. PMID- 11856628 TI - Human masseter inhibitory reflexes evoked by repetitive electrical stimulation. AB - OBJECTIVES: The relationship between the masseter inhibitory reflex (MIR) and nociceptive processing in the trigeminal region was studied in 10 healthy subjects. Based on the known increase in perceived sensory intensity following repetitive stimulation of the nociceptive system, we examined the hypothesis that the MIR reflects noxious activity in the trigeminal system by determining the possible relation between changes in MIR and perceived sensory intensity. METHODS: The MIR was quantitated and compared with psychophysical measures following repetitive peri-oral electrical stimulation (5 square wave pulses of 0.5ms repeated at 2Hz). In addition to the early (ES1) and late (ES2) periods of exteroceptive suppression, two periods with apparent excitation could be distinguished from the background electromyografical activity: (a) the inter suppression period (ISP) between the ES1 and the ES2, and (b) the post suppression period (PSP) after the ES2. A computer algorithm was used to detect and quantitate ES1, ES2, ISP, and PSP. The response variables were (a) onset latencies and (b) magnitudes of suppression (ES1 and ES2) and excitation (ISP and PSP). RESULTS: Consistent reduction of the magnitude of ES2 suppression in response to repetitive stimuli was observed below as well as above the pain detection threshold. CONCLUSIONS: The observed reduction of the magnitude of ES2 suppression is not specifically related to nociceptive processing. Habituation or net inhibitory effects on inhibitory pre-motor neurones (i.e. disinhibition) are possible mechanisms for the observed reduction of the magnitude of ES2 suppression after repetitive stimulation. PMID- 11856629 TI - L-dopa effects on preprogramming and control activity in a skilled motor act in Parkinson's disease. AB - OBJECTIVES: The authors investigated whether preprogramming (Bereitschaftspotential, BP) and control activity (skilled performance positivity, SPP) in a bimanual, sequential skilled performance task (SPT) is sensitive to L-dopa administration in non-demented Parkinson's disease (PD) patients. METHODS: Movement related potentials (MRPs) were recorded in 12 non demented parkinsonian patients before and after acute L-dopa administration, and in 17 control subjects, all of whom were performing SPT for the first time. BP, SPP and correct performances were evaluated both as a grand average and in sequential blocks in order to verify the learning effect. RESULTS: After L-dopa administration the PD patients scored a significantly higher percentage of correct performances (P<0.05), linked to a decreased BP amplitude (P<0.001) and an increased SPP amplitude (P<0.005), than before therapy. Dynamic evaluation through the block analysis did not show any learning effect in off-therapy patients but showed that L-dopa intake improved learning, linked to a BP amplitude decrease (P<0.005) and a SPP amplitude increase (P<0.05). Furthermore, L-dopa minimized differences in the learning trend between off-therapy PD patients and controls. CONCLUSIONS: Our findings suggest that skilled motor learning is impaired in non-demented untreated PD patients. Dopaminergic drug administration seems to restore the ability of PD patients to use more automatic motor strategies, as demonstrated by the electrophysiological and behavioural pattern, which became more similar to that of normal subjects. PMID- 11856630 TI - The effects of alteration of effector and side of movement on movement-related cortical potentials. AB - The amplitude of the movement-related cortical potential (MRCP) preceding self paced voluntary movements is larger if subjects alter between flexions of two fingers compared with repetitive movements of the same finger. However, earlier studies were confined to alternating movements between limbs only and therefore could not differentiate effects of between-limbs from within-limb alteration. The present study was designed to examine effects of alteration of finger (from index to middle, and vice versa) and hand (from left to right, and vice versa) independently from each other. MRCPs were recorded in 20 right-handed healthy young subjects with electrodes placed at Fp1, Fp2, F7, F3, F4, F8, T7, C5, C3, C1, C2, C4, C6, T8, P7, P3, P4, P8, O1 and O2. Subjects made self-paced flexion movements with the index or middle finger of each hand by pressing one out of 4 response buttons. In the alternating conditions, subjects pressed two buttons in a strictly alternating fashion. Every finger was combined with every other finger, arriving at a total of 6 alternating conditions. In the 4 regular conditions, subjects pressed a particular button repetitively across trials. The results show the following: (1) MRCPs over contralateral sensorimotor areas are higher if subjects change the side of movement than if the movements are done with the same hand repeatedly. (2) MRCPs over lateral parietal areas are higher for ipsilateral than contralateral movements in the regular conditions but also increase contralaterally after a change of the side of movement. (3) Any change of side or finger is associated with a widespread increase of negativity over the left hemisphere. PMID- 11856631 TI - Intra- and interindividual variability of motor responses to repetitive transcranial magnetic stimulation. AB - OBJECTIVES: Repetitive transcranial magnetic stimulation (rTMS) can modify cortical excitability and is widely used for clinical and research purposes. We sought to determine the intra- and interindividual variability of its effects on motor cortex excitability, and whether repeated paired-pulses yield less variability than repeated single-pulses. METHODS: We investigated rTMS over the left motor cortex of 6 healthy subjects and recorded motor evoked potentials (MEPs) from the right abductor digiti minimi muscle. Eighty single suprathreshold stimuli or conditioning-test pairs of stimuli were delivered at 2Hz frequency. The pairs consisted of a subthreshold pulse followed by a suprathreshold pulse after 2, 5 or 10ms. In each subject we studied all types of rTMS 5 times on separate days. Single suprathreshold pulses at 0.17Hz preceded rTMS for baseline determination. RESULTS: The day-to-day variability of MEPs during either type of rTMS was small compared to the subject-to-subject variability. MEPs increased during all types of rTMS except for interstimulus interval (ISI) 2ms. Paired pulses yielded less variability than single-pulse rTMS. CONCLUSIONS: Motor responses to rTMS show a high interindividual, but a low intraindividual variability. Repeated paired-pulses yield less variability than repeated single pulses. PMID- 11856632 TI - Perceptual and motor contributions to performance and ERP components after incorrect motor activation in a flanker reaction task. AB - OBJECTIVES: The aims of this study are to evaluate contributions of response and perceptual processes to reaction performance in a flanker reaction task, and to investigate whether event-related potential (ERP) component N2 and error negativity N(e) represent similar or functionally distinct cortical mechanisms. METHODS: ERPs, lateralized readiness potentials (LRPs) and reaction performance were measured in a flanker task with arrows as targets and congruent or incongruent flankers. Squares were used as neutral flankers. Target color signaled a response of the hand indicated by (PRO) or against (ANTI) the target arrow's pointing direction. RESULTS: On both PRO and ANTI conditions, performance was facilitated by congruent and impaired by incongruent flankers. In the ERPs on trials with late response errors an N2 was evident before an N(e). In addition, ERPs on correct trials showed an N2 particularly after incongruent flankers on PRO but for each flanker type on ANTI conditions. On incongruent ANTI trials, two successive response conflicts occurred but only a single N2 appeared. CONCLUSIONS: The results indicate that differences in perceptual processing contribute significantly to the flanker effects on task performance and provide further evidence that N2 and N(e) represent different cortical mechanisms. The data also suggest that N2 is not a real-time correlate of incorrect response suppression. PMID- 11856633 TI - Single motor axon conduction velocities of human upper and lower limb motor units. A study with transcranial electrical stimulation. AB - OBJECTIVES: To calculate conduction velocities (CV) of single motor axons innervating hand, forearm and leg muscles, weak anodal electrical transcranial stimuli were used and single motor unit potentials were recorded in 17 normal subjects. METHODS: The central motor conduction time and neuromuscular transmission delay were subtracted from the latency of unit response to cortical stimulation and single motor axon CV were calculated. RESULTS: In extensor indicis proprius (EIP) units, CV ranged from 30.3 to 76.1m/s (mean: 51.3 +/- 7.1m/s, 139 units). In first dorsal interosseous (FDI), they ranged from 45.1 to 66.2m/s (mean: 54.6 +/- 2.6m/s, 88 units). In tibialis anterior (TA), velocities ranged from 27.8 to 55.9m/s (mean: 41.3 +/- 7.5m/s, 123 units). In FDI units, velocities were compared with those obtained with the F-wave method (range: 50.3 64.5m/s, mean: 58.1 +/- 2.0m/s). CONCLUSIONS: Compared with previously published values, the present method gives better access to slow-conducting units, first recruited by transcranial stimulation and voluntary effort. The spectrum of individual CV was much broader for EIP and TA than for FDI. A linear decline of maximal CV with age was observed, while minimal CV were not affected, suggesting that aging causes a selective loss of the fastest-conducting units. PMID- 11856634 TI - Reciprocal Ia inhibition in patients with asymmetric spinal spasticity. AB - OBJECTIVES: Previous studies on reciprocal Ia inhibition in spinal spasticity were contradictory, probably due to differences in the etiology, severity, and the course of recovery from the disease. The purpose of the present study was to establish a correlation between Ia inhibition and clinical abnormalities in spasticity following a spinal cord injury (SCI). METHODS: We studied reciprocal Ia inhibition in bilateral soleus muscles in five SCI patients with marked asymmetry of spasticity and functional recovery for the right and left legs. Reciprocal Ia inhibition was determined from the short latency suppression of the soleus H-reflex by conditioning stimulation of the peroneal nerve. RESULTS: In all the patients, Ia inhibition was asymmetric. Ia inhibition in the legs with good recovery and less spasticity was pronounced, but Ia inhibition in the more spastic legs was small or absent. Facilitation was seen only on the sides with poor recovery in two patients. In the healthy subjects the amount of Ia inhibition varied, but no side-to-side difference was found. CONCLUSIONS: These results suggest that reciprocal Ia inhibition varies according to the functional recovery. Pronounced Ia inhibition may be related to good functional recovery in patients with SCI. PMID- 11856635 TI - Pain-related magnetic fields evoked by intra-epidermal electrical stimulation in humans. AB - OBJECTIVES: We recently developed a new method for the preferential stimulation of Adelta fibers in humans. The aim of the present study was to examine whether this method can serve as an appropriate stimulus in a magnetoencephalographic study. METHODS: We recorded somatosensory-evoked magnetic fields (SEFs) following intra-epidermal electrical stimulation applied to the hand and elbow. Superficial parts of the skin were electrically stimulated through a needle electrode whose tip was inserted in the epidermis. RESULTS: In all 13 subjects, the equivalent current dipole was estimated in the secondary somatosensory cortices (SII). In 5 out of 13 subjects, simultaneous activation of the primary somatosensory cortex (SI) in the hemisphere contralateral to the stimulation was identified. The mean peak latencies of magnetic fields corresponding to contralateral SI, SII and ipsilateral SII activation following hand stimulation were 162, 158 and 171 ms, respectively. The respective latency following elbow stimulation was 137, 139 and 157 ms, respectively. Estimated peripheral conduction velocity was 15.6m/s. CONCLUSIONS: All the results were consistent with previous findings in pain SEF studies. We concluded that our novel intra-epidermal electrical stimulation is useful for pain SEF studies since it does not need special equipment and is easy to control. PMID- 11856636 TI - Galvanic-evoked myogenic responses in patients with an absence of click-evoked vestibulo-collic reflexes. AB - OBJECTIVES: To show that galvanic-evoked responses on the sternocleidomastoid muscle (SCM) are useful for differentiating labyrinthine lesions from retro labyrinthine lesions in patients with an absence of click-evoked vestibulo-collic reflexes. METHODS: We studied the average responses in the unrectified electromyographic (EMG) activities of the SCM to galvanic stimulation (3mA, 1ms). The cathodal electrode was on the mastoid, and the anodal electrode was on the forehead. Twenty-two healthy subjects and 28 patients with vestibular disorders were studied. All of the 28 patients showed the unilateral absence of vestibulo collic reflexes evoked by 95dBnHL clicks on the affected side. RESULTS: In healthy subjects mastoid-forehead galvanic stimulation produced a positive negative biphasic EMG response at short latency on the SCM ipsilateral to the cathodal electrode. All patients with labyrinthine lesions showed biphasic EMG responses even in the affected side. In contrast, almost all patients with retro labyrinthine lesions (16/18) showed no response or a decreased response on the affected side. CONCLUSIONS: These results suggest that galvanic-evoked myogenic responses on the SCM may be useful in the differential diagnosis of labyrinthine lesions from retro-labyrinthine lesions in patients with an absence of vestibulo collic reflexes evoked by clicks. PMID- 11856637 TI - Properties of human skin mechanoreceptors in peripheral neuropathy. AB - OBJECTIVES: To investigate the properties of mechanoreceptors in patients with peripheral neuropathy. The skin mechanoreceptor is a terminal organ of the primary sensory neuron, which is likely to be affected earlier and more severely than is the nerve trunk by peripheral neuropathies. METHODS: Single sensory unit responses to air-puff and electric stimulation were recorded using the microneurographic technique in the glabrous skin of the hand. Receptor transduction time was estimated by a latency difference between electric- and air puff-induced responses. RESULTS: A total of 38 mechanoreceptive units were obtained from 14 normal subjects. All the units responded to air-puff stimuli irrespective of the receptor type, and receptor transduction time was approximately 2 ms. A total of 32 units were recorded from 11 patients with neuropathy of variable causes. Seven (22%) of the 32 neuropathic units did not respond to air-puffs despite their ability to respond to electric stimulation. Compared to normal ones, units from patients with peripheral neuropathy had significantly higher mechanical thresholds, but receptor transduction times did not differ significantly. CONCLUSIONS: Changes in receptor properties in human neuropathy are characterized by increased mechanical threshold without prolongation of receptor transduction time, possibly due to a high threshold for generating receptor potentials. PMID- 11856641 TI - Characteristics of in vitro osteoblastic cell loading models. AB - Normal loading strains of 200-2000 (mu)epsilon to bone result in bending forces, generating mechanical stretch and pressure gradients in canaliculi that drive extracellular fluid flow, resulting in stress on the membranes of osteocytes, lining cells, and osteoblasts. Under excess loading, as well as during unloading (e.g., microgravity, bed rest), the fluid shift and resultant change in interstitial fluid flow may play a larger role in bone remodeling than mechanical stretch. The in vitro model systems used to investigate mechanical loading of bone generate either fluid shear, hydrostatic compression, biaxial stretch, uniaxial stretch, or a combination of two or more of these forces. The results of in vitro experiments suggest that fluid shear is a major factor affecting bone cell metabolism. Both the flow-loop apparatus (which produces pulsatile flow and uses fluid shear as its principal stimulus) and the uniaxial silicone plate stretching apparatus (which generates cyclic stretch) create a reproducible and consistent stimulus. Endpoints measured in flow experiments, however, are short term and usually short lived, and it is unknown whether these changes impact the function of differentiated osteoblasts. Endpoints measured in uniaxial stretch experiments are generally long-term-sustained effects of mechanical perturbation and more easily relatable to changes in osteoblastic activity. Biaxial stretch devices create both bending and compressive forces, resulting in different types of force on the cells, with the relative amount of each depending on the position of the cell in the device. Therefore, systems that incorporate pulsatile fluid flow or uniaxial stretch as the principal stimulus should be further developed and implemented in the study of the relationship between mechanical loading and bone response. PMID- 11856642 TI - Abnormal osteoclast morphology and bone remodeling in a murine model of a lysosomal storage disease. AB - Mucopolysaccharidosis type VII (MPS VII) is a heritable lysosomal storage disease caused by a deficiency in beta-glucuronidase (GUSB) activity, leading to progressive accumulation of undegraded glycosaminoglycans in many tissues. Clinical features include growth and mental retardation, hearing and visual defects, shortened lifespan, and skeletal deformities. A murine model of MPS VII has been described that shares many of the manifestations of the human disease, including the skeletal dysplasia. In this study we describe abnormalities in the cellular morphology and function of osteoclasts and a localized defect in bone formation rate in the MPS VII mouse. Ultrastructural analysis revealed that MPS VII osteoclasts fail to form ruffled border membranes and many appeared to be detached from the bone surface. Following bone marrow transplantation, osteoclasts derived from wild-type donors showed normal morphology and were closely associated with the bone surface in MPS VII recipients. In vitro bone resorption assays demonstrated that MPS VII osteoclasts formed significantly smaller and fewer pits than those formed by osteoclasts derived from normal mice of the same strain. Although osteoclast morphology and function appeared to be abnormal in the MPS VII mouse, interleukin-1 (IL-1)-induced osteoclastogenesis in vivo was not affected. In addition to the osteoclast defects, MPS VII mice demonstrated a slower rate of bone matrix deposition in the epiphysis by in vivo calcein labeling experiments. These data suggest that abnormal morphology and function of MPS VII osteoclasts, combined with deficient matrix deposition, may contribute to the skeletal defects observed in this lysosomal storage disease. PMID- 11856640 TI - Involvement of receptor activator of NFkappaB ligand and tumor necrosis factor alpha in bone destruction in rheumatoid arthritis. AB - Bone loss represents a major unsolved problem in rheumatoid arthritis (RA). The skeletal complications of RA consist of focal bone erosions and periarticular osteoporosis at sites of active inflammation, and generalized bone loss with reduced bone mass. New evidence indicates that osteoclasts are key mediators of all forms of bone loss in RA. TNF-alpha is one of the most potent osteoclastogenic cytokines produced in inflammation and is pivotal in the pathogenesis of RA. Production of tumor necrosis factor-alpha (TNF-alpha) and other proinflammatory cytokines in RA is largely CD4(+) T-cell dependent and mostly a result of interferon-gamma (IFN-gamma) secretion. Synovial T cells contribute to synovitis by secreting IFN-gamma and interleukin (IL)-17 as well as directly interacting with macrophages and fibroblasts through cell-to-cell contact mechanisms. Activated synovial T cells express both membrane-bound and soluble forms of receptor activator of NF-kappaB ligand (RANKL). In rheumatoid synovium, fibroblasts also provide an abundant source of RANKL. Furthermore, TNF alpha and IL-1 target stromal-osteoblastic cells to increase IL-6, IL-11, and parathyroid hormone-related protein (PTHrP) production as well as expression of RANKL. In the presence of permissive levels of RANKL, TNF-alpha acts directly to stimulate osteoclast differentiation of macrophages and myeloid progenitor cells. In addition, TNF-alpha induces IL-1 release by synovial fibroblasts and macrophages, and IL-1, together with RANKL, is a major survival and activation signal for nascent osteoclasts. Consequently, TNF-alpha and IL-1, acting in concert with RANKL, can powerfully promote osteoclast recruitment, activation, and osteolysis in RA. The most convincing support for this hypothesis has come from in vivo studies of animal models. Protection of bone in the presence of continued inflammation in arthritic rats treated with osteoprotegerin (OPG) supports the concept that osteoclasts mediate bone loss, providing further evidence that OPG protects bone integrity by downregulating osteoclastogenesis and promoting osteoclast apoptosis. Modulation of the RANKL/OPG equilibrium in arthritis may provide additional skeletal benefits, such as chondroprotection. The nexus between T-cell activation, TNF-alpha overproduction, and the RANKL/OPG/RANK ligand-receptor system points to a unifying paradigm for the entire spectrum of skeletal pathology in RA. Strategies that address osteoclastic bone resorption will represent an important new facet of therapy for RA. PMID- 11856643 TI - Effect of etidronate on bone in orchidectomized and sciatic neurectomized adult rats. AB - The purpose of the present study was to determine whether etidronate treatment could prevent bone loss caused by orchidectomy (ORX) and unilateral sciatic neurectomy (NX) in adult male rats. Seventy-four male Wistar rats, aged 10 months, were randomly divided into eight groups: baseline controls (n = 10); age matched sham-operated controls (AMC; n = 9); ORX (n = 9); NX (n = 10); ORX + NX (n = 9); ORX + etidronate treatment (ORX + E; n = 7); NX + E (n = 10); and ORX + NX + E (n = 10). Etidronate treatment (10 mg/kg per day subcutaneously) was initiated 2 weeks after surgery and was continued for 2 weeks. Four weeks after surgery, bone mineral density (BMD) of the proximal and middle tibia (PT and MT, respectively), distal and middle femur (DF and MF, respectively), and fourth lumbar vertebral body (LVB) was measured by dual-energy X-ray absorptiometry (Model DCS-600, Aloka, Tokyo, Japan). The mechanical properties of the MF and third LVB were measured by three-point bending and compression tests, respectively. Levels of urinary deoxypyridinoline (Dpd) and serum osteocalcin (Oc) were also measured by enzyme-linked immunosorbent assay. Four weeks of aging had no significant effects on BMD, bone mechanical properties, or bone markers. ORX significantly increased the levels of urinary Dpd and serum Oc, which resulted in significant decreases in BMD of the PT, MT, DF, MF, and fourth LVB, as well as the mechanical strength (maximum load) of the MF and third LVB. NX significantly increased levels of urinary Dpd and decreased levels of serum Oc, resulting in a significant decrease in BMD of the PT, DF, and fourth LVB. The ORX induced decrease in BMD of the PT was more pronounced when combined with NX. Etidronate treatment for NX, ORX, and ORX + NX rats significantly decreased levels of urinary Dpd and serum Oc, resulting in complete prevention of loss of BMD and/or bone mechanical strength. The present study demonstrates the efficacy of etidronate treatment for prevention of bone loss caused by testosterone deficiency and immobilization in adult male rats. PMID- 11856639 TI - Misconceptions (1): epiphyseal fusion causes cessation of growth. PMID- 11856644 TI - The selective estrogen receptor modulator raloxifene regulates osteoclast and osteoblast activity in vitro. AB - Raloxifene is a selective estrogen receptor modulator (SERM) that prevents bone loss. Although it is largely used for the treatment of osteoporosis, the mechanisms by which this compound modulates the activity of bone cells are still poorly understood. In this study we investigate whether raloxifene affects osteoclast and osteoblast activity in vitro. Bone marrow cultures were established from neonatal mice and treated with 1,25(OH)(2) vitamin D(3) (VitD(3), 10(-8) mol/L) to induce osteoclast generation. Similar to 17beta estradiol, raloxifene significantly reduced the number of osteoclasts in a concentration-dependent manner, with maximal inhibition at 10(-11) mol/L (-48%). However, as for 17beta-estradiol, at a high concentration (10(-7) mol/L), the inhibitory effect of raloxifene was abolished. In a pit assay, raloxifene inhibited bone resorption. A maximal effect was observed at 10(-9) mol/L, and maintained at a high concentration, indicating that inhibition of osteoclast formation and inhibition of bone resorption may be due to activation of, at least in part, different pathways. Osteoblasts from neonatal mice calvariae were also exposed to raloxifene. In these cells, this compound induced a concentration dependent increase of proliferation, which was blocked by the estrogen-receptor antagonist ICI 164,384. Raloxifene also increased the osteoblast-specific transcription factor Cbfa1/Runx2 and alpha2 procollagen type I chain mRNAs, with a pattern that only partially coincided with that of 17beta-estradiol. Consistent with decreased osteoclastogenesis, raloxifene inhibited the mRNA expression of interleukin (IL)-1beta and IL-6 at a low concentration, but not at a high concentration, whereas 17beta-estradiol had similar effects on IL-6 and inhibited IL-1beta at both concentrations. Furthermore, both compounds were able to inhibit tumor necrosis factor (TNF)-alpha-induced IL-1beta, but not IL-6, increase. In conclusion, these data show that raloxifene negatively modulates osteoclasts, and positively affects osteoblasts, suggesting not only an antiresorptive role, but also an osteoblast stimulatory role. PMID- 11856645 TI - Investigation of osteocalcin, osteonectin, and dentin sialophosphoprotein in developing human teeth. AB - Biochemical investigations in rodents have shown that numerous mineralized matrix proteins share expression in bone, dentin, and cementum. Little information is available regarding the expression pattern of these proteins in human tissues, particularly during tooth formation. The aim of this study was to identify the expression pattern of the two major noncollagenous proteins of bone and dentin, osteocalcin (OC) and osteonectin (ON), in comparison to the dentin-specific protein, dentin sialophosphoprotein (DSPP). Mandibles from fetuses (5-26 weeks), neonate autopsies, forming teeth from 10-12-year-old patients, third molars extracted for orthodontic reasons, and bone tumors were collected with approval from the National Ethics Committee. Human OC, ON, and DSPP mRNAs were detected by reverse transcription-polymerase chain reaction (RT-PCR) in fetal mandibles (5-11 weeks) and in primary cell cultures of dental pulp. In addition, OC, ON, and DSPP proteins were localized in forming human mineralized tissues using immunohistochemistry. In vivo, DSPP expression was associated with tooth terminal epithelial-mesenchymal interaction events, amelogenesis and dentinogenesis. Transient DSPP expression was seen in the presecretory ameloblasts with continuous expression in the odontoblasts. In contrast, both osteoblasts and odontoblasts showed a temporal gap between OC and ON expression in early development. ON was expressed in the initial stages of cytodifferentiation, whereas OC was expressed only during the later stages, especially in the teeth. At the maturation stage of enamel formation, both proteins were detected in odontoblasts and their processes within the extracellular matrix. In contrast to bone, OC was not localized extracellularly within the collagen-rich dentin matrix (predentin or intertubular dentin), but was found in the mature enamel. ON was present mostly in the nonmineralized predentin. These results demonstrate for the first time that both OC and ON are produced by human odontoblasts and determine the expression pattern of DSPP in human teeth, and suggest that OC and ON move inside the canalicule via odontoblast cell processes becoming localized to specific extracellular compartments during dentin and enamel formation. These distinct extracellular patterns may be related to the nature of DSPP, OC, and ON interactions with other matrix-specific macromolecules (i.e., amelogenin, dentin matrix protein-1) and/or to the polarized organization of odontoblast secretion as compared with osteoblasts. PMID- 11856646 TI - Comparisons between bone and cementum compositions and the possible basis for their layered appearances. AB - In humans, age estimation from the adult skeleton represents an attempt to determine chronological age based on growth and maturational events. In teeth, such events can be characterized by appositional growth layers in midroot cementum. The purpose of this study was to determine the underlying cause of the layered microstructure of human midroot cementum. Whether cementum growth layers are caused by changes in relative mineralization, collagen packing and/or orientation, or by variations in organic matrix apposition was investigated by subjecting midroot sections of human canine teeth to analysis using polarized light and scanning electron microscopy (SEM). Polarized light was used to examine transverse midroot sections in both mineralized and demineralized states. Mineralized sections were also reexamined following subsequent decollagenization. Polarized light was additionally used in the examination of mineralized sections taken transversely, longitudinally, and obliquely from the same tooth root. From the birefringence patterns it was concluded that collagen orientation does not change with varying section plane. Instead, the mineral phase was most responsible for the birefringence of the cementum. SEM studies suggested that neither collagen packing nor collagen orientation change across the width of the cementum, confirming and validating the results of the polarized light examination. Also, SEM analysis using electron backscatter and the electron probe suggested no changes in the mean atomic number density, calcium, phosphate, and sulfur levels across the width of the cementum. Therefore, we conclude that crystalline orientation and/or size is responsible for the layered appearance of cementum. PMID- 11856648 TI - Is clinical expressiveness of Paget's disease of bone decreasing? AB - Recent data have suggested secular changes implying a current trend toward decreased clinical severity of Paget's disease of bone (PD). To test this hypothesis, we conducted a study comparing the characteristics of two groups of PD patients, as disclosed from a sample assessed systematically. The investigation was a hospital-based study of all cases followed up at our unit since 1980. Throughout the follow-up period, diagnosis was based on standard X ray criteria and the same clinical assessment was applied. Group I (n = 124) represented patients born before 1926, whereas group II (n = 109) included those born after that year. A bone scan performed with 99mTc-EHDP was available for all patients. X-rays of the pelvis and spine, and views of any hot spot observed on the scintigraphy scans were reviewed. The skeletal extent of PD, based on bone scan uptake, was determined by using the index proposed by Coutris. Alkaline phosphatase and hydroxyproline excretion levels were determined in blood and urine, respectively. Baseline characteristics were recorded on a purpose-designed computerized database. The proportion of males (47% in group I vs. 65% in group II; p = 0.007) and the mean (+/-SD) age at diagnosis (69.0 +/- 8.15 vs. 54.3 +/ 9.14; p < 0.001) differed significantly between groups. The year of birth showed a strong negative correlation with age at diagnosis (r = -0.83, p < 0.0001) and a weak, but significant, negative correlation with extent of bone lesion (r = 0.20; p = 0.002). Likewise, subjects born prior to 1926 showed a greater percentage of affected skeleton cases (9.6 plus minus 8.01 vs. 7.06 +/- 5.79; p = 0.001). Group I individuals who had pelvic and/or femoral bone lesions were more prone to suffer "pagetic coxopathy" (65% vs. 40%; p = 0.003) with "protrusio acetabuli" (32% vs. 17%; p = 0.01), and the percentage of patients showing radiographic Monckeberg-type vascular calcifications (36% vs. 14%; p = 0.0006) was higher than in those born after 1926. No other epidemiologically clinically, or biochemically relevant differences were seen in the crude analysis. Multivariate analysis identified extent of skeletal lesions (OR = 0.76; p = 0.01), age at diagnosis (OR = 0.79; p = 0.008), number of bones involved (OR = 1.53; p = 0.03), and occupation (p < 0.0001) as the predictive variables linked to year of birth. Our data are consistent with a temporal tendency toward a smaller number of bone lesions and a decreased percentage of instances of affected skeleton. An earlier age at recent diagnosis times and absence of any relevant clinical or biochemical differences seems more likely linked to recent changes in referral and sociological patterns. PMID- 11856647 TI - 2-methoxyestradiol induces interferon gene expression and apoptosis in osteosarcoma cells. AB - 2-Methoxyestradiol (2-ME), a naturally occurring mammalian metabolite of 17beta estradiol, has been implicated as a physiological inhibitor of tumor cell proliferation. In this study, the effects of 2-ME on cultured osteosarcomatous cells were investigated. Dose-dependent growth inhibition was observed in MG63 and TE85 human osteosarcoma cells exposed to 2-ME. The cell killing by 2-ME was ligand-specific; the immediate precursor (2-hydroxyestradiol), the parent compound (17beta-estradiol), and the equivalent metabolite of estrone (2 methoxyestrone) exhibited less potency and efficacy. Furthermore, 2-ME was similarly effective at killing immortalized human fetal osteoblastic cells (hFOB) with and without estrogen receptor-alpha and -beta and rat osteosarcoma cells (ROS17/2.8). The cytotoxicity of 2-ME was selective to transformed and immortalized osteoblastic cells; 2-ME (2 microm) had no effect on the proliferation of primary cultures of human osteoblasts. Co-treatment with the potent estrogen receptor ligand, ICI-182,780, did not reduce 2-ME-induced osteosarcoma cell death, implying that this action is not mediated by conventional estrogen receptors. The expression levels of bone matrix protein genes, type 1 collagen and osteonectin, were transiently reduced after 2-ME treatment, suggesting that the surviving cells are capable of producing bone matrix. The 2-ME-mediated killing of osteosarcoma cells was due to the induction of apoptosis; treatment induced expression of interferon genes within 12 h and histological evidence of apoptosis within 48 h of 2-ME treatment. Thus, our results demonstrate that 2-ME is highly cytotoxic to osteosarcoma cells but not normal osteoblasts. These findings suggest that further study of 2-ME as a potential intervention for treatment of osteosarcoma is warranted. PMID- 11856649 TI - Mechanical consequence of trabecular bone loss and its treatment: a three dimensional model simulation. AB - Age-related changes in the microstructure of trabecular bone, such as decreases in trabecular number and trabecular thickness, lead to reductions in mechanical properties, such as Young's modulus and strength. Current drug therapy, such as bisphosphonate or parathyroid hormone, improves the mechanical properties of bone mainly by increasing the trabecular thickness, but not increasing the trabecular number. However, the mechanical efficacy of these treatments has not been fully quantified using trabecular bone models. In this study, we used an idealized three-dimensional (3D) microstructural model of trabecular bone to create bone loss either through trabeculae thinning or random removal of trabeculae, and simulated treatment by increasing the trabeculae thickness of the remaining trabeculae. The reduction in either the Young's modulus or the strength due to trabeculae loss was proportional to a much higher power of reduction in bone volume fraction than due to trabeculae thinning. This indicates that bone loss due to trabeculae loss is much more detrimental to Young's modulus and strength of trabecular bone than due to trabeculae thinning, indicating the importance of trabecular number and connectivity in the mechanical integrity of trabecular bone. In general, treatments by increasing the trabecular thickness of remaining trabeculae after trabeculae loss cannot fully recover the initial mechanical properties of intact bone, even if bone loss is fully recovered, whereas trabecular thickening can fully restore the mechanical properties after bone loss by trabeculae thinning. The results also show that the residual loss in mechanical properties is dependent on the extent of trabeculae loss. PMID- 11856650 TI - Age- and gender-related differences in bone mineral status and biochemical markers of bone metabolism in Northern Chinese men and women. AB - The aim of this study was to provide insight into the bone mineral status and biochemical markers of bone metabolism in a Chinese population from Shenyang, in the north of China, where hip fracture incidence is low. A total of 194 healthy men and women, aged 25-35 years and 65-75 years, were studied. Bone mineral density (BMD) at the lumbar spine (LS) and femoral neck (FN) was measured using dual-energy X-ray absorptiometry. Fasting blood and 24 h urine samples were collected for bone alkaline phosphatase (bAP), osteocalcin, calcium, phosphate, and free deoxypyridinoline (Dpd). Both older women and men had lower BMD compared with younger women and men by 27.2 +/- 3.0% (mean +/- SE) and 9.8 +/- 3.0% at the LS and 22.0 +/- 3.4% and 12.8 +/- 3.4% at the FN, respectively, after adjusting for bone and body size (p < 0.01). BMD at the two sites was lower in older women than in older men by 10.7 +/- 4.1% and 10.2 +/- 4.2%, respectively, after size correction (p < 0.05). Plasma bAP, osteocalcin, calcium, and phosphate concentrations were higher in older women than younger women by 69.3 +/- 9.7%, 77.2 +/- 11.1%, 7.5 +/- 2.3%, and 8.0 +/- 3.8%, respectively, and older men by 67.6 +/- 11.1%, 72.1 +/- 11.0%, 7.7 +/- 2.3%, and 23.8 +/- 3.8%, respectively (p < 0.01). However, plasma osteocalcin, calcium, and phosphate concentrations were lower in older men compared with their younger counterparts by 35.6 +/- 11.0%, 8.7 +/- 2.3%, and 14.1 +/- 3.8%, respectively (p < 0.05). Urinary calcium and phosphate output were lower in older men compared with younger men by 48.0 plus minus 10.3% and 27.6 +/- 6.9%, respectively (p < 0.01), whereas there were no differences between older and younger women. No differences in Dpd between older and younger groups were found. This study demonstrates that bone mineral status is lower in older people in Shenyang, as has been shown in populations elsewhere. The pronounced low bone mineral status in older women may be associated with increased bone turnover, which was not observed in older men. PMID- 11856651 TI - Modifiable determinants of bone status in young women. AB - The purpose of this study was to evaluate the contributions of exercise, fitness, body composition, and calcium intake during adolescence to peak bone mineral density and bone structural measurements in young women. University Hospital and 75 healthy, white females in the longitudinal Penn State Young Women's Health Study were included. Body composition, total body, and hip bone mineral density (BMD) were measured by dual-energy X-ray absorptiometry (DXA), exercise scores by sports-exercise questionnaire during ages 12-18 years, and estimated aerobic capacity by bike ergometry. Section modulus values (a measurement of bending strength) cross-sectional area (CSA), subperiosteal width, and cortical thickness were calculated from DXA scan data for the femoral neck and femoral shaft. Calcium intakes were calculated from 39 days of prospective food records collected at 13 timepoints between ages 12 and 20 years; supplemental calcium intakes were included. Section moduli at the femoral neck and shaft were correlated significantly with lean body mass, sports-exercise scores (R(2) = 0.07 0.19, p < 0.05), and aerobic capacity (R(2) = 0.06-0.57, p < 0.05). Sports exercise scores correlated with BMD at the femoral neck and shaft. Average total daily calcium intake at age 12-20 years ranged from 486 to 1958 mg/day and was not significantly associated with total or regional peak BMD or bone structure measures at 20 years of age. It was shown that achievable levels of exercise and fitness have a favorable effect on BMD and section modulus of the femoral neck and femoral shaft in young adult women, whereas daily calcium intake of >500 mg in female adolescents appears to have little, if any effect. PMID- 11856653 TI - Hypercorticism blunts circadian variations of osteocalcin regardless of nutritional status. AB - Anorexia nervosa (AN) and Cushing's syndrome (CS) are both responsible for osteoporosis. The mechanisms leading to osteoporosis in AN include hypogonadism, nutritional depletion, and in some cases hypercorticism. Osteocalcin circulating level is a serum marker of osteoblastic activity that follows a circadian rhythm (OCR). Serum osteocalcin is decreased in both CS and AN and can be increased with treatment. In this study we analyzed the influence of combined cortisol and nutritional status on osteocalcin levels and its circadian rhythm in these two different models of hypercorticism, one nutritionally replete (CS) and one nutritionally deplete (AN), and we evaluated the effects of their treatment (surgical cure and weight gain, respectively). Before treatment, osteocalcin levels were lower in CS (n = 16) and AN (n = 42) than in controls and in the AN patient subgroup with hypercorticism (n = 13) compared to those without (n = 29). OCR was absent in CS and in AN patients with hypercorticism, whereas their circadian cortisol cycle was maintained. In CS, successful surgical treatment increased osteocalcin levels (n = 5) and restored OCR. In AN, weight gain (n = 13) induced a significant decrease in cortisol levels in hypercortisolic AN patients, and restored normal osteocalcin levels and OCR. In conclusion, we found that hypercorticism was associated with a decrease in osteocalcin levels in nutritionally replete or deplete patients and that OCR was more affected by cortisol levels than by cortisol cycle. PMID- 11856652 TI - Effects of vitamin D analog, 22-oxa-1,25-dihydroxyvitamin D(3), on bone reconstruction by vascularized bone allograft. AB - We previously reported that vascularized bone allograft using immunosuppressants, such as cyclosporine A (CsA), is one approach for reconstruction of large bone defects in both experimental animals (Microsurgery 15:663; 1994) and clinically in humans (Lancet 347:970, 1996). Because immunosuppressive agents such as CsA induce significant side effects, including bone loss, other therapeutic agents supporting successful vascularized bone allografts have been sought after. We investigated the effects of 22-oxa-1,25-dihydroxyvitamin D(3) (OCT) on vascularized bone allograft, and compared its effects with CsA. Twelve-week-old DA rats with the major histocompatibility antigen (MHC) RT-1(a) were used as donors and age-matched Lewis rats with MHC RT-1(l) used as recipients. Allografted bones in rats treated with vehicle were rejected completely. Soft X ray examination demonstrated that administration of OCT (0.5 microg/kg per day) for 12 weeks after bone graft induced bone union as effective as treatment for 12 weeks with CsA (10 mg/kg per day). Transplanted bones in OCT-treated rats showed higher bone mineral density than that in CsA-treated rats. Histologically, transplanted bones in OCT-treated rats at 12 weeks were nonvital, but these bones united with recipient vital bones. After cessation of 12 week treatment with OCT, new bone formation occurred around the grafted nonvital bones during a 9 month period. Transplanted bones in CsA-treated rats were vital and formed union with recipient bones, whereas cortical bones became thin when compared with nonvital bones in OCT-treated rats. Urinary deoxypyridinoline levels in rats treated with CsA were significantly higher than levels in rats treated with OCT, suggesting accelerated bone resorption in CsA-treated rats. These results suggest that OCT exerts an anabolic action on bone reconstruction by allogeneic bone transplantation. PMID- 11856654 TI - Novel mutations in the a3 subunit of vacuolar H(+)-adenosine triphosphatase in a Japanese patient with infantile malignant osteopetrosis. AB - A case of infantile malignant osteopetrosis is described. The patient died from respiratory hemorrhage at 7 months of age despite treatment that included high doses of active vitamin D and administration of interferon-gamma. A postmortem examination revealed the presence of many osteoclasts in the bone, which lacked ruffled borders. This observation was consistent with the histology of bone reported in Atp6i-knockout mice, which lack the gene encoding the a3 subunit of vacuolar-type H(+)-adenosine triphosphatase (ATPase). Sequence analysis of the TCIRG1 gene encoding the a3 subunit revealed two novel mutations: a deletion/insertion mutation in exon 9 and a T-to-C transition at the splice donor site of intron 19. The former mutation caused a frame shift and premature stop codon. The latter was associated with abnormal splicing, which was confirmed by sequencing the products amplified by reverse transcription-polymerase chain reaction (RT-PCR), using total RNA from the liver specimen as template. Although several mutations in the TCIRG1 gene in infantile malignant osteopetrosis have been reported in other populations, this is the first case of a Japanese patient with a mutation identified in this gene. These results support the important role of the subunit in the function of the proton pump. PMID- 11856655 TI - Premenstrual syndrome and premenstrual dysphoric disorder. AB - It is almost impossible to cover all the options available for the treatment of premenstrual syndrome and premenstrual dysphoric disorder. This article discusses the most common and relevant options. PMID- 11856656 TI - Psychiatric issues related to infertility, reproductive technologies, and abortion. AB - Infertility, treatment with reproductive technologies, and abortion are among the most emotionally weighty and philosophically contentious experiences in most patients' lives. They involve the most intimate body parts and behaviors and the most heartfelt hopes and profound disappointments. They can strain relationships with partners, relatives, and friends. The primary care practitioner who is informed about the psychological impact of these experiences can play an essential role in interpreting medical information; helping patients think through their own values, resources, and options; facilitating communication between members of a couple and with their friends and family; providing emotional support; and identifying and treating psychiatric disorders that sometimes occur before, during, or after these experiences. PMID- 11856657 TI - Postpartum disorders in primary care: diagnosis and treatment. AB - Postpartum psychiatric illness consists of a highly prevalent group of disorders, which can result in serious dysfunction and require treatment. Patients are more likely to seek help for these disorders from their primary care physicians rather than mental health professionals. While severe postpartum depression and psychoses are easily recognized, milder or more insidious forms of depressive illness frequently are missed. Heightened sensitivity to and screening for the presence of these prevalent psychiatric disorders facilitates and enhances recognition of postpartum disorders and leads to more expeditious treatment. PMID- 11856658 TI - Menopause and perimenopause: the role of ovarian hormones in common neuroendocrine syndromes in primary care. AB - The climacteric and menopause are characterized by erratic fluctuations and decline in ovarian steroid hormones that have broad impact on physical and psychological phenomena leading to a number of clinical syndromes encountered in primary care settings. This endocrine transition is abrupt in women and is not often adequately assessed using objective measures, leading to potential overuse of various medications to treat the physiological and psychological consequences of decline in ovarian hormones. This article reviews such issues and explains how primary care physicians may incorporate the endocrine changes more effectively in their evaluation and treatment of midlife women. PMID- 11856660 TI - Eating disorders: a guide for the primary care physician. AB - The diagnostic criteria for eating disorders are described, including assessment of complications related to semi-starvation, binge eating, and purging. The key components of treatment are presented, including normalization of eating, individual and family therapy, and cognitive behavioral therapy. The role of the primary care physician in diagnosis and treatment is outlined. PMID- 11856659 TI - Depression and anxiety in older women. AB - The recommended shift in paradigm for assessment and treatment of depression and anxiety in the primary care setting includes a more holistic medical care approach, one that pays attention to the patient's mental health status and her functional level of social role recovery in addition to symptom relief. Practice Guidelines of professional specialities should be expanded to include attention to initializing mental health care in primary care practice and parameters for early referral and, if indicated, later follow-up. Our medical education system, at all levels, needs to become considerably more inclusive of issues of aging, gender, and mental health. Ongoing attention must be given to the health care cost burden of under recognition and under treatment of anxiety and depression, alleviation of stigma, treatment to functional recovery, and alleviation of caregiver burden. PMID- 11856661 TI - Body dysmorphic disorder: a guide for primary care physicians. AB - Body dysmorphic disorder (BDD), a preoccupation with an imagined or slight defect in appearance, is a relatively common psychiatric disorder that often presents to nonpsychiatric physicians. Body dysmorphic disorder is associated with marked impairment in functioning, notably poor quality of life, and a high suicide attempt rate. Most patients seek and receive surgery or nonpsychiatric medical or dermatologic treatment, often with a perceived poor outcome despite an objectively acceptable result. In contrast, psychiatric treatment, serotonin reuptake inhibitors and cognitive-behavioral therapy, often are effective. This clinically focused review describes the clinical features and prevalence of BDD, the disorder's treatment response, how to recognize and diagnose BDD, and practical suggestions for primary care physicians who encounter these often difficult-to-treat patients. PMID- 11856662 TI - Approaching sexual issues in primary care. AB - This article outlines the typical phases in the development and maintenance of healthy female sexual functioning, and highlights the problems related to sexual functioning that women may present in a primary care practice. The author discusses an approach to the sexual problems of women subjected to trauma and those with certain psychiatric illnesses, and the effect of psychotropic medications on sexual functioning. PMID- 11856663 TI - Primary care and victims of domestic violence. AB - Providing quality health care involves integrating routine inquiry about domestic violence into ongoing clinical practice. This means asking all women patients, and others who may be at risk, about abuse in their lives. Whether or not a woman chooses to use services or leave her partner, our intervention is very important. Some women return to violent partners several times before they feel safe enough to leave, feel they can survive on their own, or can accept that the person they love will not change. Make sure that she has follow-up for her medical problems and appropriate referrals for mental health and substance abuse problems when indicated. PMID- 11856664 TI - Posttraumatic stress disorder in women: assessment and treatment in primary care. AB - The focus of this article is to provide primary care clinicians with a better understanding of women who have undergone sexual trauma and have related post traumatic stress disorder. Victimization has adverse physical and mental health effects and affects a woman's clinical presentation, her coping skills, and the primary care intervention strategies needed to treat her. The article reviews issues of victimization and related PTSD among women, including the prevalence and sequel of victimization, and provides a theoretical framework for primary care intervention, treatment, and referral. PMID- 11856667 TI - Primary prevention and practical techniques to encourage mental wellness. AB - Women patients must be made aware that they need to train and perhaps readjust their attitudes and minds to handle and improve their mental wellness. Being their own person, with awareness of whom and how they want to be, strengthens their sense of self and simplifies their lives with awareness and understanding of the choices unique to them as individuals. PMID- 11856665 TI - The role of sex hormones in psychopathology: focus on schizophrenia. AB - The example of schizophrenia is used to illustrate how sex hormones affect the presentation of illness and its treatment. Organization and activation effects of hormones are explained, and behavior is shown to result from a complex interplay of hormones, brain mechanisms, and social pressures. Sex differences in schizophrenia (in onset age, symptoms, antipsychotic, and other treatment) are consequences of this interplay and impact on the clinician's ability to diagnose, treat, prognosticate, and prevent the disability and distress of schizophrenia. PMID- 11856666 TI - The role of complementary and alternative therapies in women's mental health. AB - The advent of the newly evolving field of Integrative Medicine has provided women with an opportunity to expand the health care resources available to prevent and treat mental health conditions. Aided by evidence-based science, practitioners can now broaden the scope of their therapeutic armamentarium and allow women to use the best of what western and complementary medicine has to offer in the field of mental health care. Acupuncture, medication, herbalism, and guided imagery are included in a list of therapeutic modalities designed to augment traditional care. It is clear that by providing such comprehensive care, a woman's physical and mental wellness and her longevity are optimized. PMID- 11856669 TI - Impact factor: the numbers game. PMID- 11856668 TI - New perspectives on the psychotherapy of women: cost effective and clinical interventions. AB - Psychotherapy is a cost-effective treatment modality with demonstrated effectiveness in various medical and psychiatric conditions. The author reviews contemporary trends in the psychodynamic understanding of and practice of psychotherapy for women. Emphasis is placed on practical interventions that the primary care clinician can use in practice. PMID- 11856670 TI - High-resolution breath-hold contrast-enhanced MR angiography of the entire carotid circulation. AB - OBJECTIVE: The purpose of this study was to evaluate the effect of breathing on image quality of the aortic arch and carotid vessels during contrast-enhanced MR angiography and to show that high-resolution breath-hold contrast-enhanced MR angiography combined with a timing-bolus technique can produce high-quality images of the entire carotid circulation. MATERIALS AND METHODS: Forty patients underwent high-resolution contrast-enhanced MR angiography on a 1.5-T Magnetom Symphony. A coronal three-dimensional (3D) gradient-echo sequence (TR/TE, 4.36/1.64; flip angle, 25 degrees) with asymmetric k-space acquisition was used. The 136 x 512 matrix yielded voxel sizes of 1.33 x 0.64 x 1.0 mm. A timing-bolus acquisition, orientated in the coronal plane to include the aortic arch, was obtained initially during free-breathing. Twenty milliliters of gadopenetate dimeglumine was injected at 2 mL/sec. Unenhanced and enhanced 3D volumes were recorded. A subtracted 3D set was calculated and subjected to a maximum-intensity projection algorithm. Half of the patients held their breath during angiography and the other half did not. Aortic arch motion was measured on the timing-bolus acquisition as the distance moved by a single pixel in both the x and y directions. Maximum-intensity-projection MR images were assessed independently by two observers, and vessel sharpness was scored on a scale of 1-5. Sharpness was also assessed quantitatively by generating a signal intensity profile across the aortic arch vessel wall and calculating the average of the upslope and downslope at full-width half maximum. Visualization of carotid branch vessels was scored on a scale of 0-5, and venous contamination was scored on a scale of 0-3. RESULTS: Average in-plane aortic arch movement was 10.3 mm in the x direction and 8.7 mm in the y direction. Quantitative and qualitative sharpness of the aortic arch and great vessel origins was better (p < 0.05) during breath-holding than during non breath-holding. No difference in the sharpness of the carotid vessels was noted between the two groups. Carotid branch vessels were well visualized from the aortic arch to the intracerebral circulation. The average venous contamination score was 0.56. CONCLUSION: Breath-holding greatly improves the sharpness of the aortic arch and great vessel origins but has no effect on visualization of the carotid vessels. High-resolution breath-hold contrast-enhanced MR angiography can produce high-quality, artifact-free images of the entire carotid circulation from the aortic arch to the intracerebral circulation. PMID- 11856672 TI - Blood flow volume quantification of cerebral ischemia: comparison of three noninvasive imaging techniques of carotid and vertebral arteries. AB - OBJECTIVE: Determination of blood flow volume is useful in assessing ischemic cerebrovascular disease. We compared the blood flow volume measurement of three noninvasive imaging techniques, namely color velocity imaging quantification, spectral Doppler imaging quantification, and MR phase-contrast flow quantification, to see how well the flow values determined by each technique agreed with one another. SUBJECTS AND METHODS: Flow volume quantification was tested experimentally using a flow simulator and by the three techniques in the vertebral and internal carotid arteries of 40 patients with histories of cerebral ischemia. In the flow simulation study, the flow values in each technique were compared with the phantom flow by the Wilcoxon's signed rank test. In the patient study, the flow values between each paired technique were compared by paired t test. The significance level was taken at p less than 0.05. RESULTS: Flow volumes were measured by color velocity imaging quantification. MR phase-contrast flow quantification agreed with the phantom flow simulation within the tested range, and spectral Doppler imaging quantification values were significantly overestimated. In patients, a large variation of the blood flow volume was obtained between each technique (p < 0.05). Among them, spectral Doppler imaging quantification showed the highest flow values in the vessels (internal carotid arteries, 312.6 mL/min; vertebral arteries, 112.0 mL/min), followed by color velocity imaging quantification (internal carotid arteries, 216.8 mL/min; vertebral arteries, 58.1 mL/min) and MR phase-contrast flow quantification (internal carotid arteries, 169.1 mL/min; vertebral arteries, 66.5 mL/min). CONCLUSION: Blood flow volume measurements determined by the three noninvasive imaging techniques on the same vessel can differ widely, and spectral Doppler imaging quantification consistently overestimated the flow volume. It is, therefore, essential that the same technique, preferably color velocity imaging quantification or MR phase-contrast flow quantification, be used for clinical follow-up investigations in the future. PMID- 11856673 TI - Wooden foreign bodies: imaging appearance. AB - OBJECTIVE: The purpose of this study was to identify the characteristic imaging features of wooden foreign bodies. MATERIALS AND METHODS: The imaging studies of 12 patients with surgically confirmed wooden foreign bodies were retrospectively reviewed. The study group consisted of seven females and five males, 10-65 years old (mean age, 36 years). All patients underwent radiography. Nine patients were evaluated with sonography, eight with MR imaging, three with CT, and one with CT arthrography. Gadolinium-enhanced MR imaging was performed in six patients. Three patients presented with a draining sinus and nine with painful swelling. Only three patients presented with a history of penetrating injury. RESULTS: Lesions were located in the foot (n = 4), hand (n = 3), thigh (n = 2), calf (n = 2), and elbow (n = 1). Radiographs failed to reveal the retained foreign bodies in all patients. With MR imaging, wooden foreign bodies displayed a variable signal intensity that was equal to or less than that of skeletal muscle on both T1- and T2-weighted images. MR imaging showed the surrounding inflammatory response in all patients. CT showed the retained wood as linear cylindric foci of increased attenuation. Wood was highly echogenic and revealed pronounced acoustic shadowing on sonography. Arthrography in one patient showed an associated reactive synovitis. CONCLUSION: The imaging appearance of wooden foreign bodies is variable; however, imaging can be quite specific, and when taken in the appropriate clinical setting, the imaging should reliably suggest the diagnosis. Sonography is frequently underused but proved most useful for the evaluation of retained wooden foreign bodies. PMID- 11856674 TI - Work flow redesign: the key to success when using PACS. AB - OBJECTIVE: Our objective is to emphasize the importance of work flow redesign, rather than filmless operation itself, to achieve cost reduction and improvement in productivity with picture archiving and communication systems (PACS). CONCLUSION: Our 8-year experience with PACS shows that the greatest benefit of the transition to a digital system has been the ability to use it as a tool to reengineer overall work flow, both in the imaging department and throughout the health care enterprise. PMID- 11856675 TI - The infected or substance abuse-impaired radiologist. PMID- 11856676 TI - Cervical spine injuries in patients 65 years old and older: epidemiologic analysis regarding the effects of age and injury mechanism on distribution, type, and stability of injuries. AB - OBJECTIVE: Our objective was to describe types and distribution of cervical spine injuries in elderly patients in regard to causative trauma mechanism and patient age. MATERIALS AND METHODS: The distribution and type of 225 cervical spine injuries in 149 consecutive patients 65 years old and older over a 5-year interval were retrospectively assessed. For each patient, initial admission imaging studies were reviewed, and injuries were classified. Trauma mechanism (falls from standing or seated height vs higher energy mechanisms) and initial clinical and neurologic status were recorded. Data were correlated according to patients' age (65-75 years and >75 years) and causative trauma mechanism. RESULTS: Ninety-five (64%) of 149 patients had upper cervical spine injuries. Fifty-nine (40%) of 149 patients had multilevel injuries. Main causes for cervical spine injuries were motor vehicle crashes in "young elderly" (65-75 years old; 36/59, 61%) and falls from standing or seated height in "old elderly" (>75 years old; 36/90, 40%). Fracture patterns at risk for neurologic deterioration were common (>50%), even in the absence of acute myelopathy or radiculopathy. Patients older than 75 years, independent of causative mechanism, and patients who fell from standing height, independent of age, were more likely to have injuries of the upper cervical spine (p = 0.026 and p = 0.006, respectively). CONCLUSION: Cervical spine injuries in elderly patients tend to involve more than one level with consistent clinical instability and commonly occur at the atlantoaxial complex. Old elderly patients and patients who fall from standing height are more prone to injuries of the upper cervical spine. PMID- 11856677 TI - Usefulness of two indirect MR imaging signs to diagnose lateral meniscal tears. AB - OBJECTIVE: We determined whether using two recently described indirect MR imaging signs would improve the sensitivity of diagnosis of lateral meniscal tears. MATERIALS AND METHODS: We identified 121 consecutive patients who had undergone knee MR imaging and knee arthroscopy. Their MR imaging examinations were evaluated for the conventional criteria of a meniscal tear (meniscal distortion or intrameniscal signal contacting the surface) and the two new signs (presence of an abnormal popliteomeniscal fascicle and posterolateral pericapsular edema). These observations were correlated with the arthroscopic findings, which were used as the gold standard. RESULTS: Thirty-two (89%) of the 36 torn lateral menisci had two or more images with distortion or signal contacting the surface. Three torn menisci and eight intact menisci had one image with distortion or surface signal. Only one of 75 menisci without distortion or surface signal was torn. An abnormal superior fascicle was highly associated (p < 0.001) with lateral meniscal tears but was not specific for a tear because three of the 14 menisci with abnormal fascicles were not torn. Posterolateral pericapsular edema was not associated with a lateral meniscal tear (p = 0.06). Using an abnormal fascicle as an additional criterion improved the sensitivity from 89% to 94%, but the difference was not statistically significant. CONCLUSION: We confirmed that an abnormal fascicle is highly associated with a lateral meniscal tear but found that posterolateral pericapsular edema was not associated with lateral meniscal tears. Identifying an abnormal fascicle did not significantly improve the sensitivity of diagnosis of a lateral meniscal tear. PMID- 11856678 TI - MR evaluation of the "arcuate" sign of posterolateral knee instability. AB - OBJECTIVE: The purpose of this study was to evaluate associated knee injuries using MR imaging in patients with the "arcuate" sign, a term referring to avulsion fracture of the proximal fibula on conventional radiographs. MATERIAL AND METHODS: MR imaging of 18 cases (17 patients, both knees in one patient) with the arcuate sign on conventional radiographs was retrospectively interpreted to evaluate the associated meniscal, ligamentous, and bony injuries. In 12 cases, MR findings were correlated with surgical results. RESULTS: In all cases, avulsed bony fragments from the proximal pole of the fibula were attached to the fibular collateral ligament, the biceps femoris tendon, or both. Tear of the posterolateral capsule was seen in 12 cases (67%). Injury of the cruciate ligaments was noted in 16 cases (89%): injury to both the anterior cruciate ligament and posterior cruciate ligament was seen in nine cases (50%), injury to only the anterior cruciate ligament was seen in four, and injury to the posterior cruciate ligament only was noted in three. Bone bruises or gross fractures were seen in all cases: bone bruises on the anteromedial femoral condyle were noted in nine cases (50%) and were seen on the anteromedial tibial condyle in five cases (28%). Tear of the medial meniscus was seen in five cases (28%) and tear of the lateral meniscus in four cases (22%). Injury to the popliteus was seen in six cases (33%). Joint effusion was associated in all cases. CONCLUSION: MR imaging is useful for evaluation of associated soft-tissue injuries in patients with the arcuate sign on conventional radiographs. Avulsion injury to the proximal fibula is an important indicator of the internal derangement of the knee and for predicting the mechanism of an injury with varus stress. Cruciate ligament tear and bone bruises on the anteromedial condyle of the femur and tibia are common associated findings. PMID- 11856680 TI - Neoplastic and tumorlike lesions detected on MR imaging of the knee in patients with suspected internal derangement: Part 2, articular and juxtaarticular entities. PMID- 11856682 TI - Anteromedial impingement of the ankle: using MR arthrography to assess the anteromedial recess. AB - OBJECTIVE: Our objective was to describe the appearance of the anteromedial tibiotalar joint on MR arthrography in patients with clinically and arthroscopically confirmed anteromedial impingement. CONCLUSION: Anteromedial impingement of the ankle is now being recognized in the orthopedic literature as a distinct entity. MR arthrographic findings of anteromedial impingement include capsular and synovial soft-tissue thickening anterior to the tibiotalar ligaments and any associated osseous abnormality. Although anteromedial impingement is uncommon compared with other impingement syndromes of the ankle, the radiologist should be aware of the diagnosis and possible findings on cross-sectional imaging. PMID- 11856679 TI - Neoplastic and tumorlike lesions detected on MR imaging of the knee in patients with suspected internal derangement: Part I, intraosseous entities. PMID- 11856683 TI - Is soft-tissue inflammation in pedal infection contained by fascial planes? MR analysis of compartmental involvement in 115 feet. AB - OBJECTIVE: The purpose of this study was to analyze compartmental involvement and patterns of spread of soft-tissue inflammation in pedal infection. MATERIALS AND METHODS: We reviewed 115 contrast-enhanced 1.5-T MR examinations of the foot in 41 women and 74 men with a mean age of 58.4 years who had undergone bone biopsy or surgery for suspected osteomyelitis. Presence of inflammation (contrast enhancement, fat signal intensity loss on T1-weighted images, and high signal intensity on T2-weighted images) was noted by two musculoskeletal radiologists in the following foot compartments: toes, medial, central, lateral, interosseous, dorsal, hindfoot, malleoli, and lower leg. Proximal and distal extension of soft tissue inflammation was analyzed. The compartment closest to the ulcer that showed MR signs of direct contiguous infection was designated the primarily infected compartment. RESULTS: Spread of inflammation across fascial planes into neighboring compartments originated from the following primary compartments: medial (3/10, 30%), central (7/16, 44%), and lateral (16/20, 80%). Spread from the hindfoot and malleoli into adjacent compartments was seen in only 7% of such cases (2/24). Inflammation from toe infections spread in 34% of cases to forefoot compartments (15/44). Inflammation from forefoot or toe infections spread in 4.5% of cases to the midfoot and in 2% of cases to the hindfoot; ascension into the calf was rare (1% of cases). Spread of inflammation into neighboring compartments was not correlated with the presence of diabetes (p = 0.81) or with osteomyelitis (p = 0.34). CONCLUSION: Soft-tissue inflammation of the forefoot tends to spread into neighboring compartments, with little respect for fascial planes. Hindfoot inflammation tends to stay confined. Spread from the foot to the lower leg is rare. PMID- 11856686 TI - Using a peptide inhibitor of the glycoprotein IIb/IIIa platelet receptor: initial experience in patients with acute peripheral arterial occlusions. AB - OBJECTIVE: The purpose of this study was to evaluate the efficacy of eptifibatide, an inhibitor of the glycoprotein (GP) IIb/IIIa platelet receptor, in the thrombolytic treatment of patients with acute peripheral arterial occlusive disease. MATERIALS AND METHODS: We retrospectively reviewed our experience with the use of a GP IIb/IIIa receptor inhibitor, eptifibatide, during thrombolysis in 17 patients with acute lower extremity arterial occlusions who also received intraarterial recombinant tissue plasminogen activator (rt-PA) and heparin. Four of the 17 patients received their loading dose of eptifibatide by direct intraarterial injection, whereas the remaining 13 received an IV loading dose. We compared their results with those of 11 other patients who received only rt-PA and heparin with respect to success and complication rates, duration of thrombolytic therapy, and total rt-PA dose. RESULTS: We found no significant difference in successful outcome (p = 1.00), major complications (p = 1.00), duration of therapy (p = 0.21), or total rt-PA dose (p = 0.67) between those who received eptifibatide and those who did not during thrombolytic therapy. However, those patients who received an intraarterial loading dose of eptifibatide required substantially less rt-PA (9.0 +/- 4.4 mg vs 38.9 +/- 30.7 mg) to achieve successful thrombolysis. CONCLUSION: The adjunctive use of a GP IIb/IIIa platelet receptor inhibitor during thrombolysis for arterial occlusions may decrease the total dose of rt-PA required for thrombolysis without compromising success or complication rates. A prospective randomized study is needed to confirm that inhibitors of the GP IIb/IIIa platelet receptor can facilitate thrombolytic therapy in patients with acute lower extremity arterial occlusions. PMID- 11856684 TI - Osteosarcoma arising in a desmoplastic fibroma of the proximal tibia. PMID- 11856688 TI - Development of a perigraft seroma around modified Blalock-Taussig shunts: imaging evaluation. AB - OBJECTIVE: The modified Blalock-Taussig shunt is a synthetic shunt between the subclavian and pulmonary artery, frequently used in the treatment of children with pulmonary hypoperfusion caused by congenital heart disease. The development of a perigraft seroma is a known complication of this procedure. We sought to describe the imaging features of a perigraft seroma and to define an optimal diagnostic strategy in patients with a suspected perigraft seroma. MATERIALS AND METHODS: Between January 1993 and December 1998, 96 children underwent 105 modified Blalock-Taussig shunt procedures. In eight children, 11 cases of perigraft seromas were identified. The mean age of these children at the time of operation was 3 years (range, 6 days to 5 years 8 months). Pre- and postoperative chest radiographs were routinely performed in the children in whom seromas had been found. Additional postoperative radiologic investigations consisted of thoracic sonography (in 11 cases), CT (in eight cases), and MR imaging (in two cases). In all cases of perigraft seroma, the modified Blalock-Taussig shunts were constructed through a posterolateral thoracotomy at the fourth intercostal space. RESULTS: On average, the chest radiographs showed the first signs of the seroma on day 10 after the surgery (range, day 1-day 30). Using thoracic sonography, it was possible to visualize the perigraft seroma and the modified Blalock-Taussig shunt in eight (73%) of 11 cases. CT and MR imaging performed equally well in revealing perigraft seromas. CONCLUSION: As was found in these critically ill children, sonography has an advantage over CT and MR imaging because of its portability and, therefore, capability for bedside use. We recommend the use of sonography as the initial imaging modality in suspected cases of perigraft seroma development. PMID- 11856690 TI - MR angiographic findings in patients with aortic endoprostheses. PMID- 11856687 TI - Evaluation of abdominal aortic aneurysm after endoluminal treatment: comparison of color Doppler sonography with biphasic helical CT. AB - OBJECTIVE: The purpose of this study is to compare color Doppler sonography with biphasic helical CT in the evaluation of abdominal aortic aneurysms after endovascular repair. MATERIALS AND METHODS: Fifty-five patients prospectively underwent both color Doppler sonography and helical CT within 7 days after treatment by endovascular stent-graft. Aneurysmal thrombosis, the patency of the grafts, and the presence of a leak were evaluated in all patients. When a perigraft leak was observed, an attempt was made to identify its origin and outflow vessels. Helical CT was considered the gold standard technique. RESULTS: Helical CT revealed aneurysmal thrombosis in 33 patients and a perigraft leak in 22 patients. In five patients, helical CT detected a small perigraft leak not shown by color Doppler sonography. In three patients with suboptimal examinations, color Doppler sonography revealed a suspected perigraft leak that was not confirmed by helical CT. In these eight patients, the perigraft leak was sealed or no longer observed during follow-up. Compared with enhanced helical CT, the sensitivity and specificity of color Doppler sonography for the diagnosis of a perigraft leak were 77% and 90%, respectively. In seven other patients, helical CT was superior to color Doppler sonography in detecting the origin of the perigraft leak and the outflow vessels. Two iliac artery dissections and one distal migration of the prosthesis were revealed only by helical CT. CONCLUSION: Although color Doppler sonography may detect substantial perigraft leaks, helical CT is superior for detecting the origin of the perigraft leak, the outflow vessels, and the detection of complications related to the procedure. PMID- 11856691 TI - The role of interventional radiology in the treatment of mediastinal collections caused by esophageal anastomotic leaks. PMID- 11856689 TI - Hemodynamic changes after self-expandable metallic stent therapy for vena cava syndrome. AB - OBJECTIVE: We examined changes in hemodynamics after self-expandable metallic stent placement in the vena cava. CONCLUSION: The rapid increase in venous return immediately after expandable metallic stent placement influenced the hemodynamics of the circulatory system. PMID- 11856692 TI - Percutaneous deployment of a low-profile bifurcated stent-graft. PMID- 11856693 TI - Usefulness of an artificial neural network for differentiating benign from malignant pulmonary nodules on high-resolution CT: evaluation with receiver operating characteristic analysis. AB - OBJECTIVE: The purpose of our study was to use an artificial neural network to differentiate benign from malignant pulmonary nodules on high-resolution CT findings and to evaluate the effect of artificial neural network output on the performance of radiologists using receiver operating characteristic analysis. MATERIALS AND METHODS: We selected 155 cases with pulmonary nodules less than 3 cm (99 malignant nodules and 56 benign nodules). An artificial neural network was used to distinguish benign from malignant nodules on the basis of seven clinical parameters and 16 radiologic findings that were extracted by attending radiologists using subjective rating scales. In the observer test, 12 radiologists (four attending radiologists, four radiology fellows, and four radiology residents) were presented with high-resolution CT images, first without and then with the artificial neural network output. Observer performance was evaluated by means of receiver operating characteristic analysis using a continuous rating scale. RESULTS: The artificial neural network showed a high performance in differentiating benign from malignant pulmonary nodules (A(z) = 0.951). The average A(z) value for all radiologists increased by a statistically significant level, from 0.831 to 0.959, with the use of the artificial neural network output. CONCLUSION: Our computerized scheme using the artificial neural network can improve the diagnostic accuracy of radiologists who are differentiating benign from malignant pulmonary nodules on high-resolution CT. PMID- 11856694 TI - Pulmonary venous infarction after radiofrequency ablation for atrial fibrillation. PMID- 11856696 TI - One operation after percutaneous diagnosis of nonpalpable breast cancer: frequency and associated factors. AB - OBJECTIVE: The purpose of this study was to determine the frequency of and factors associated with performing one therapeutic operation after percutaneous diagnosis of nonpalpable breast cancer. MATERIALS AND METHODS: Retrospective review was performed of records of 350 consecutive women who had therapeutic surgery after percutaneous imaging-guided core biopsy diagnosis of nonpalpable breast cancer. Records were reviewed to determine the frequency of performing one operation and associated factors. Statistical analysis was performed. RESULTS: One operation was performed in 283 (80.9%) of 350 women, including 106 (95.5%) of 111 women who had mastectomy and 177 (74.1%) of 239 women who had breast conserving surgery. At bivariate analysis, one operation was significantly more likely in women who had no underestimation (p < 0.001), mastectomy rather than breast conservation (p < 0.001), axillary dissection during the first operation (p < 0.001), percutaneous diagnosis of infiltrating carcinoma (p = 0.001), or mammographic mass (p = 0.006). At multivariate analysis, one operation was significantly more likely if underestimation was absent (odds ratio [OR] = 10.1, 95% confidence interval [CI] = 4.2-24.7) or if mastectomy was performed (OR = 8.7, 95% CI = 3.2-23.5); for women who had breast-conserving surgery, one operation was significantly more likely if underestimation was absent (OR = 11.4, 95% CI = 3.9-33.2) or if a mammographic mass was present (OR = 2.4, 95% CI = 1.3 4.6). CONCLUSION: One operation was performed in 80.9% of women with percutaneously proven nonpalpable breast cancer, including 74.1% of women who had breast-conserving surgery and 95.5% of women who had mastectomy. Among women who had breast conservation, one operation was significantly more likely if histologic underestimation was absent or if a mammographic mass was present. PMID- 11856695 TI - Prediction of postoperative lung function in patients with lung cancer: comparison of quantitative CT with perfusion scintigraphy. AB - OBJECTIVE: Prediction of postoperative lung function is important in preoperative evaluation of patients with lung cancer. Perfusion scintigraphy is the current method to assess the fractional contribution of lung function of the remaining lung. We developed a quantitative CT method and compared it with perfusion scintigraphy for predictions of postoperative forced expiratory volume in 1 sec (FEV1) in patients with lung cancer. SUBJECTS AND METHODS: Forty-four patients with lung cancer undergoing lung resection with preoperative CT and perfusion scintigraphy were enrolled. Quantitative CT used a dual threshold (-500 and -910 H) on standard preoperative CT to semiautomatically extract lung volume without emphysema or tumor and atelectasis, which we defined as "functional lung volume." Prediction was calculated from preoperative FEV1 multiplied by the fractional contribution of functional lung volume of the remaining lung by quantitative CT. Perfusion scintigraphy was the standard method. Predictions were correlated with postoperatively measured FEV1. RESULTS: Both quantitative CT and perfusion scintigraphy predicted postoperative FEV1 well in patients who underwent pneumonectomy (n = 28, r = 0.88 vs r = 0.86) and in lobectomy (n = 16, r = 0.90 vs r = 0.80) (both, p < 0.001). There was good agreement between the two methods by the Bland-Altman method. In the four patients with low measured postoperative FEV1 (<40% predicted normal), quantitative CT had true-positive prediction in four and perfusion scintigraphy, in only two. CONCLUSION: Given its simplicity, we proposed that quantitative CT be widely used in predicting postoperative FEV1. PMID- 11856698 TI - Interpreting data from audits when screening and diagnostic mammography outcomes are combined. AB - OBJECTIVE: The objective of this study was to use mathematic models to aid mammography practices in interpreting outcomes data derived from a combination of screening and diagnostic examinations, and in interpreting diagnostic mammography outcomes data that are not segregated by indication for examination. MATERIALS AND METHODS: We analyzed outcomes from 51,805 consecutive mammography examinations. Screening and diagnostic examinations were audited separately. Diagnostic examinations were audited by indication for examination. Extrapolating from our known mix of screening (79%) and diagnostic (21%) examinations, we determined expected combined outcomes for various mixes that might be encountered in clinical practice. Similarly, we determined the expected overall diagnostic mammography outcomes for various clinically relevant mixes of indications for examination. RESULTS: Outcomes vary substantially depending on the mix of screening and diagnostic examinations performed. For example, expected outcomes for practices with screening-diagnostic mixes of 90-10% and 50-50% are, respectively: rate of abnormal findings, 6% versus 11%; rate of positive biopsy findings, 38% versus 42%; cancer detection rate, 10 per 1,000 versus 30 per 1,000; mean invasive cancer size, 14.4 mm versus 16.0 mm; nodal metastasis rate, 8% versus 11%; and rate of stage 0 and stage I cancers, 87% versus 82%. Diagnostic outcomes also vary substantially according to indication for examination, with a higher rate of abnormal findings, a higher rate of positive biopsy findings, and a larger mean invasive cancer size expected for mixes involving a high percentage of workups for palpable lesions. CONCLUSION: When screening and diagnostic mammography outcomes are not segregated during auditing, and when diagnostic outcomes are not segregated by indication for examination, analysis of combined audit data should be based on extrapolations from known outcomes. PMID- 11856699 TI - New sonographic criteria for recommending fine-needle aspiration biopsy of nonpalpable solid nodules of the thyroid. AB - OBJECTIVE: The purpose of our study was to provide new sonographic criteria for fine-needle aspiration biopsy of nonpalpable solid thyroid nodules. MATERIALS AND METHODS: Sonographic scans of 155 nonpalpable thyroid nodules in 132 patients were prospectively classified as having positive or negative findings. Sonographic findings that suggested malignancy included microcalcifications, an irregular or microlobulated margin, marked hypoechogenicity, and a shape that was more tall than it was wide. If even one of these sonographic features was present, the nodule was classified as positive (malignant). If a nodule had none of the features described, it was classified as negative (benign). The final diagnosis of a lesion as benign (n = 106) or malignant (n = 49) was confirmed by fine-needle aspiration biopsy and follow-up (>6 months) in 83 benign nodules, by fine-needle aspiration biopsy and surgery in 44 malignant and 15 benign lesions, and by surgery alone in five malignant and eight benign lesions. The sensitivity, specificity, positive predictive value, negative predictive value, and accuracy were calculated on the basis of our proposed classification method. RESULTS: Of 82 lesions classified as positive, 46 were malignant. Of 73 lesions classified as negative, three were malignant. The sensitivity, specificity, positive predictive value, negative predictive value and accuracy based on our sonographic classification method were 93.8%, 66%, 56.1%, 95.9%, and 74.8%, respectively. CONCLUSION: Considering the high level of sensitivity of our proposed sonographic classification, fine-needle aspiration biopsy should be performed on thyroid nodules classified as positive, regardless of palpability. PMID- 11856700 TI - Cystic lymph node metastases in papillary thyroid carcinoma. AB - OBJECTIVE: The aim of this study was to illustrate and discuss the sonographic spectrum of surgically proven cystic nodal metastases from papillary thyroid carcinoma. By correlative evaluation of the sonographic imaging findings to gross pathology and histology, our purpose was to provide useful hints to differentiate cystic lymph node metastases from other benign cystic neck lesions such as branchial cysts. MATERIALS AND METHODS: Sonographic examinations of 74 patients (47 women, 27 men; mean age, 49 years) with 97 histologically confirmed cystic lymph nodes metastases from papillary thyroid carcinoma were included in the study. The anatomic relationship of the nodes relative to the primary tumor was recorded, and all cystic nodes were qualitatively categorized as either simple (purely cystic) or complex (thickened outer wall, internal nodules, internal septations, and calcifications). All imaging findings were compared with gross pathologic specimens. RESULTS: Most of the cystic metastases were ipsilateral to the primary tumor (87.8%) and located in the mid or lower jugular chain (73.2%). In 14.9% of all patients, cystic lymph node metastases were the initial manifestation of disease. Only 6.2% of all lymph node metastases were purely cystic (all of these occurred in patients less than 35 years old). Of the 91 complex metastases, a thickened outer wall was present in 35.2% of patients, internal nodules in 42.9%, and internal septations in 57.1%. No calcifications were seen in the 91 complex metastases, and two or more findings were seen in 23.1%. All sonographic findings were verified by surgery. CONCLUSION: In most of the patients, cystic lymph node metastases are characterized sonographically by the presence of a thickened outer wall, internal echoes, internal nodularity, and septations. However, in younger patients, the lymph nodes might appear purely cystic, thereby mimicking branchial cysts and thus requiring biopsy for final diagnosis and therapy planning. PMID- 11856701 TI - Percutaneous ethanol injection for treatment of cervical lymph node metastases in patients with papillary thyroid carcinoma. AB - OBJECTIVE: The objective of this study was to evaluate the technique, efficacy, and side effects of percutaneous ethanol injection in patients with limited cervical nodal metastases from papillary thyroid carcinoma. SUBJECTS AND METHODS: Fourteen patients who had undergone thyroidectomy for papillary thyroid carcinoma presented with limited nodal metastases (one to five involved nodes) in the neck between May 1993 and April 2000. All patients had received previous iodine-131 ablative therapy with a mean total dose per patient of 7,548 MBq. Ten of the patients either were considered poor surgical candidates or preferred not to have surgery, and all were unresponsive to iodine-131 therapy. Each metastatic lymph node was treated with percutaneous ethanol injection, and patients received both clinical and sonographic follow-up. RESULTS: Twenty-nine metastatic lymph nodes in our 14 patients were injected. Mean sonographic follow-up was 18 months (range, from 2 months to 6 years 5 months). All treated lymph nodes decreased in volume from a mean of 492 mm(3) before percutaneous ethanol injection to a mean volume of 76 mm(3) at 1 year and 20 mm(3) at 2 years after treatment. Six nodes were re-treated 2-12 months after initial percutaneous ethanol injection because of persistent flow on color Doppler sonography (n = 4), stable size (n = 1), or increased size (n = 1). Two patients developed four new metastatic nodes during the follow-up period that were amenable to percutaneous ethanol injection. Two patients developed innumerable metastatic nodes that precluded retreatment with percutaneous ethanol injection. No major complications occurred. All patients experienced long-term local control of metastatic lymph nodes treated by percutaneous ethanol injection. In 12 of 14 patients, percutaneous ethanol injection was successful in controlling all known metastatic adenopathy. CONCLUSION: Sonographically guided percutaneous ethanol injection is a valuable treatment option for patients with limited cervical nodal metastases from papillary thyroid cancer who are not amenable to further surgical or radioiodine therapy. PMID- 11856702 TI - Irradiated carcinoma of the tongue: correlation of MR imaging findings with pathology. AB - OBJECTIVE: MR imaging was prospectively correlated with pathologic findings to study whether MR imaging can differentiate viable from nonviable tumor tissue in the irradiated carcinoma of the tongue. SUBJECTS AND METHODS: MR examinations were performed after radiation therapy in 21 patients with carcinoma of the tongue. All patients underwent either a total glossectomy or hemiglossectomy after radiation therapy. Specimens were examined microscopically. Radiation changes were histologically graded into four groups (I, minimal cellular changes; II, presence of cellular changes and partial destruction of the tumor; III, only nonviable tumor cells; IV, no tumor cells). MR examinations included T2-weighted imaging, unenhanced T1-weighted imaging, dynamic contrast-enhanced imaging, and contrast-enhanced T1-weighted imaging. RESULTS: On unenhanced T1-weighted images, the lesion was hypointense, except for two patients with histologic grade III. On T2-weighted images, the lesion appeared hyperintense in 12 of 14 patients with viable tumor cells (grades I and II); however, the lesion was hypointense in four, and isointense in two of seven patients with nonviable tumor cells (grades III or IV). Contrast-enhanced T1-weighted images showed that the degree of contrast enhancement of the lesion was equal to or lower than that of a normal salivary gland in 18 of 21 patients. For the time of maximal enhancement of the lesion on dynamic imaging, there was no substantial difference between viable (grades I and II) and nonviable (grades III and IV) tumor tissue. CONCLUSION: The present study shows that T2-weighted imaging is feasible for differentiating viable from nonviable tumor tissue in irradiated carcinoma of the tongue. PMID- 11856703 TI - Comparison of permeability in high-grade and low-grade brain tumors using dynamic susceptibility contrast MR imaging. AB - OBJECTIVE: The purpose of this study was to compare permeability measurements in high-grade and low-grade glial neoplasms using a T2(*)-weighted method. Our hypothesis was that permeability measurements using a T2(*)-weighted technique would show permeability in high-grade neoplasms to be higher than that in low grade neoplasms. MATERIALS AND METHODS: Twelve patients with biopsy-proven high grade neoplasms and 10 patients with biopsy-proven low-grade neoplasms underwent dynamic susceptibility contrast MR perfusion imaging (TR/TE, 1500/80) after bolus infusion of 0.2 mmol/kg of MR contrast material. Color-coded permeability weighted maps were created using a model that weights relative contributions to signal intensity from intravascular T2(*) effects and extravascular T1 effects from blood-brain barrier permeability. Two measures of permeability were performed: mean value of highest permeability found on three images through the tumor (mean regional value) and highest value found at any region of interest in the tumor (single area of maximum permeability). Depending on the normality of the data sets, we used the Wilcoxon's rank sum test or the two-tailed Student's t test for statistical analysis. RESULTS: For low-grade tumors, the range was 0.006 0.041, and the median of the mean regional value for each image was 0.017. For high-grade tumors, the range was 0.005-0.092, and the median of the mean regional value was 0.035 (p = 0.025). For low-grade tumors, the range was 0.008-0.045, and the mean of the single area of maximum values was 0.02. For high-grade tumors, the range was 0.007-0.136, and the mean of the single area of maximum values was 0.054 (p = 0.018). CONCLUSION: Permeability values for high-grade tumors obtained using a T2(*)-weighted method were significantly greater than those for low-grade tumors and are consistent with previous studies reporting results using T1 weighted methods. PMID- 11856704 TI - Fibrous dysplasia involving the base of the skull. PMID- 11856705 TI - Improved pediatric multidetector body CT using a size-based color-coded format. AB - OBJECTIVE: CT technique should be adjusted while scanning infants and children. One format that has proven successful in simplifying pediatric care and reducing medical error is the size-based, color-coded Broselow-Luten pediatric system. This color-coded system can serve as a format for CT protocols. The purpose of this investigation was to compare variation (or error) occurrence and technologist preference for conventional and color-coded formats for pediatric multidetector body CT protocols. MATERIALS AND METHODS: Multidetector CT examinations were set up using either a conventional or a color-coded format for a period of 6 weeks each. Variations (errors) from protocol parameters (including tube current, detector configuration, table speed, and IV contrast media dose) were tabulated. Qualitative assessment consisted of a survey of CT technologists (n = 20) for preference in six areas related to ease of use and clarity of the formats. RESULTS: There were 44 CT examinations (n = 30 infants and children) in the conventional group and 55 CT examinations (n = 31 infants and children) in the color-coded protocol format group. Overall, the number of errors was significantly less in the color-coded group (p < 0.01), with a significantly lower error percentage in individual parameters affecting radiation dose, including tube current, detector configuration, and table speed (p < 0.05). In all areas, the color-coded format was preferred over the conventional format (p < 0.0003). CONCLUSION: Color-coded CT formatting is an extension of a clinical color-coded system. This system provides an easy, expeditious, consistent, and preferable format for general pediatric body CT protocols. Most importantly, the color-coded system can reduce variations (errors) in the radiology department. PMID- 11856706 TI - Perinatal sonographic diagnosis of cardiac fibroma with MR imaging correlation. PMID- 11856708 TI - Assessment of ventricular function with single breath-hold real-time steady-state free precession cine MR imaging. AB - OBJECTIVE: The aim of our study was to evaluate whether a recently developed real time steady-state free precession (trueFISP) cine sequence could be used to assess left ventricular function in a single breath-hold. CONCLUSION: Using real time trueFISP permits one to assess left ventricular function in a single breath hold. The dramatic reduction in data acquisition time does require some compromises. The temporal and spatial resolutions of images obtained with real time trueFISP were considerably lower than those achieved with segmented trueFISP. Further reduction of the TR or the use of sensitivity encoding could improve temporal resolution and eliminate other limitations of real-time trueFISP. PMID- 11856709 TI - CT of a renal artery pseudoaneurysm caused by a stab wound. PMID- 11856710 TI - Anovaginal and rectovaginal fistulas: endoluminal sonography versus endoluminal MR imaging. AB - OBJECTIVE: The exact location of anovaginal and rectovaginal fistulas cannot be determined by physical examination and conventional techniques. The objective of our study was to compare the accuracy of endoluminal sonography and endoluminal MR imaging in revealing the location of anovaginal and rectovaginal fistulas. MATERIALS AND METHODS: Nineteen consecutive patients (age range, 28-56 years; median age, 39 years) with clinical indications of an anovaginal or rectovaginal fistula were included in our retrospective study. Endoluminal sonography was performed using a 7.5-MHz transducer. Endoluminal MR imaging was performed at 0.5 T for 10 patients and 1.5 T for nine patients; axial T2-weighted gradient-echo, coronal and sagittal T2-weighted turbo spin-echo (0.5 T), or axial and radial T2 weighted turbo spin-echo and axial T2-weighted fat saturated turbo spin-echo (1.5 T) images were obtained. For a variety of reasons, surgery of the fistula was not attempted in six of these 19 patients. The imaging findings were compared with the findings obtained during surgery in the remaining 13 patients. RESULTS: In 12 of the 13 patients, the fistula was found during surgery: seven of the fistulas were anovaginal, and five were rectovaginal. Findings of endoluminal sonography were true-positive in 11 patients, true-negative in one, and false-negative in one. Findings of endoluminal MR imaging were true-positive in 11 patients, false negative in one, and false-positive in one. Positive predictive value for endoluminal sonography and endoluminal MR imaging were 100% and 92%, respectively. Imaging findings for anal sphincter defects were comparable. CONCLUSION: Endoluminal sonography and endoluminal MR imaging have comparable positive predictive values in revealing the location of anovaginal and rectovaginal fistulas. PMID- 11856712 TI - Xanthogranulomatous oophoritis: MR imaging findings with pathologic correlation. PMID- 11856713 TI - MR imaging of an atypical vaginal leiomyoma. PMID- 11856711 TI - Sonographic and MR imaging findings of testicular epidermoid cysts. AB - OBJECTIVE: The purpose of this study is to analyze the sonographic and MR imaging findings of testicular epidermoid cysts. CONCLUSION: Sonographic findings of a markedly heterogeneous intratesticular mass with or without alternating hypo- and hyperechoic layers surrounded by a hypoechoic or echogenic rim and the absence of flow on color Doppler sonography suggest the preoperative diagnosis of testicular epidermoid cysts. T2-weighted MR imaging findings of a high-signal-intensity mass with or without low-signal-intensity foci surrounded by a low-signal-intensity rim and the absence of enhancement on contrast-enhanced T1-weighted MR images can strengthen the preoperative diagnosis. These imaging findings can offer a basis for surgeons to attempt testis-sparing surgery instead of orchiectomy. PMID- 11856714 TI - Thick-section half-Fourier rapid acquisition with relaxation enhancement MR cholangiopancreatography: effects of i.v. administration of gadolinium chelate. AB - OBJECTIVE: Our purpose was to investigate the effects of i.v. administration of gadolinium chelate on thick-section MR cholangiopancreatography performed with half-Fourier rapid acquisition with relaxation enhancement sequence. SUBJECTS AND METHODS: Unenhanced and enhanced MR cholangiopancreatograms obtained in 50 consecutive patients were quantitatively analyzed with region-of-interest measurements and were qualitatively evaluated by three independent radiologists unaware the patient information. A phantom study was performed to verify the effects. RESULTS: The mean contrast-to-noise ratios of the gallbladder, common bile duct, and main pancreatic duct significantly increased after gadolinium chelate administration (p < 0.005). The mean depiction score of the main pancreatic duct increased significantly with one radiologist (p < 0.05) and marginally with another (p < 0.06), and the mean depiction scores of the background structures and renal pelvis significantly decreased with all three radiologists (p < 0.001). The phantom study showed the results, indicating that T2- and T2(*)-shortening effects of gadolinium chelate caused the effects in vivo. CONCLUSION: i.v. administration of gadolinium chelate improves the depiction of pancreaticobiliary ducts in some selected patients, while decreasing the depiction in others with less frequency. There may be a value of enhanced MR cholangiopancreatography when unenhanced MR cholangiopancreatography is not sufficient. PMID- 11856715 TI - FDG positron emission tomography detection of pelvic nodal metastases in gynecologic cancer. PMID- 11856716 TI - Milk of calcium on mammography. PMID- 11856717 TI - Be sure to insert--appropriately and safely--a hookwire! PMID- 11856719 TI - Premedication before i.v. contrast-enhanced CT resulting in steroid-induced psychosis. PMID- 11856718 TI - Fatal cardiac arrest during infusion of nonionic contrast media in a patient with essential thrombocythemia. PMID- 11856721 TI - Manipulation of impact factors by editors of scientific journals. PMID- 11856722 TI - Atypical meningioma spreading via the perivascular spaces. PMID- 11856723 TI - CT and scintigraphy of aggressive lymphadenopathic mastocytosis. PMID- 11856724 TI - Lumbosacral epidural abscess due to hidradenitis suppurativa. PMID- 11856725 TI - CT diagnosis of abdominal compartment syndrome. PMID- 11856727 TI - Slac2-a/melanophilin, the missing link between Rab27 and myosin Va: implications of a tripartite protein complex for melanosome transport. AB - Myosin Va is a member of the unconventional class V myosin family, and a mutation in the myosin Va gene causes pigment granule transport defects in human Griscelli syndrome and dilute mice. How myosin Va recognizes its cargo (i.e. melanosomes), however, has remained undetermined over the past decade. In this study, we discovered Slac2-a/melanophilin to be the "missing link" between myosin Va and GTP-Rab27A present in the melanosome. Deletion analysis and site-directed mutagenesis showed that the N-terminal Slp (synaptotagmin-like protein) homology domain of Slac2-a specifically binds Rab27A/B isoforms and that the C-terminal half directly binds the globular tail of myosin Va. The tripartite protein complex (Rab27A.Slac2-a.myosin Va) in melanoma cells was further confirmed by immunoprecipitation. The discovery that myosin Va indirectly recognizes its cargo through Slac2-a, a novel Rab27A/B effector, should shed light on molecular recognition of its specific cargo by class V myosin. PMID- 11856728 TI - Loss of p53 compensates for alpha v-integrin function in retinal neovascularization. AB - alpha(v)-Integrin antagonists block neovascularization in various species, whereas 20% of alpha(v)-integrin null mice are born with many normal looking blood vessels. Given that blockade of alpha(v)-integrins during angiogenesis induces p53 activity, we utilized p53 null mice to elucidate whether loss of p53 can compensate for alpha(v)-integrin function in neovascularization of the retina. Murine retinal vascularization was inhibited by systemic administration of an alpha(v)-integrin antagonist. In contrast, mice lacking p53 were refractory to this treatment, indicating that neovascularization in normal mice depends on alpha(v)-integrin-mediated suppression of p53. Blockade of alpha(v)-integrins during neovascularization resulted in an induction of p21(CIP1) in wild type and, surprisingly, in p53 null retinas, indicating that alpha(v)-integrin ligation regulates p21(CIP1) levels in a p53-independent manner. In conclusion, we demonstrate for the first time an in vivo intracellular mechanism for compensation of integrin function and that p53 and alpha(v)-integrins act in concert during retinal neovascularization. PMID- 11856730 TI - ADP-dependent glucokinase/phosphofructokinase, a novel bifunctional enzyme from the hyperthermophilic archaeon Methanococcus jannaschii. AB - A gene encoding an ADP-dependent phosphofructokinase homologue has been identified in the hyperthermophilic archaeon Methanococcus jannaschii via genome sequencing. The gene encoded a protein of 462 amino acids with a molecular weight of 53,361. The deduced amino acid sequence of the gene showed 52 and 29% identities to the ADP-dependent phosphofructokinase and glucokinase from Pyrococcus furiosus, respectively. The gene was overexpressed in Escherichia coli, and the produced enzyme was purified and characterized. To our surprise, the enzyme showed high ADP-dependent activities for both glucokinase and phosphofructokinase. A native molecular mass was estimated to be 55 kDa, and this indicates the enzyme is monomeric. The reaction rate for the phosphorylation of D glucose was almost 3 times that for D-fructose 6-phosphate. The K(m) values for D fructose 6-phosphate and D-glucose were calculated to be 0.010 and 1.6 mm, respectively. The K(m) values for ADP were 0.032 and 0.63 mm when D-glucose and D fructose 6-phosphate were used as a phosphoryl group acceptor, respectively. The gene encoding the enzyme is proposed to be an ancestral gene of an ADP-dependent phosphofructokinase and glucokinase. A gene duplication event might lead to the two enzymatic activities. PMID- 11856729 TI - Sweeping model of dynamin activity. Visualization of coupling between exocytosis and endocytosis under an evanescent wave microscope with green fluorescent proteins. AB - Vesicle recycling through exocytosis and endocytosis is mediated by a coordinated cascade of protein-protein interactions. Previously, exocytosis and endocytosis were studied separately so that the coupling between them was understood only indirectly. We focused on the coupling of these processes by observing the secretory vesicle marker synaptobrevin and the endocytotic vesicle marker dynamin I tagged with green and red fluorescent proteins under an evanescent wave microscope in pheochromocytoma cells. In control cells, many synaptobrevin expressing vesicles were found as fluorescent spots near the plasma membrane. Upon electrical stimulation, many of these vesicles showed an exocytotic response as a transient increase in fluorescence intensity followed by their disappearance. In contrast, fluorescent dynamin appeared as clusters increasing slowly in number upon stimulation. The clusters of fluorescent dynamin moved around beneath the plasma membrane for a significant distance. Simultaneous observations of green fluorescent dynamin and red fluorescent synaptobrevin indicated that more than 70% of the exocytotic responses of synaptobrevin had no immediate dynamin counterpart at the same site. From these findings it was concluded that dynamin-mediated recycling is not directly coupled to exocytosis but rather completed by a scanning movement of dynamin for the sites of invaginating membrane destined to endocytosis. PMID- 11856731 TI - Catalysis of tyrosyl-adenylate formation by the human tyrosyl-tRNA synthetase. AB - Although the active site residues in the Bacillus stearothermophilus and human tyrosyl-tRNA synthetases are largely conserved, several differences exist between the two enzymes. In particular, three amino acids that stabilize the transition state for the activation of tyrosine in B. stearothermophilus tyrosyl-tRNA synthetase (Cys-35, His-48, and Lys-233) are not present in the human enzyme. This raises the question of whether the activation energy for the tyrosine activation step is higher for the human tyrosyl-tRNA synthetase than for the B. stearothermophilus enzyme. In this paper, we demonstrate that intrinsic fluorescence changes can be used to monitor the pre-steady state kinetics of human tyrosyl-tRNA synthetase. In contrast to the B. stearothermophilus enzyme, catalysis of the tyrosine activation step is potassium-dependent in the human tyrosyl-tRNA synthetase. Specifically, potassium increases the forward rate constant for tyrosine activation 260-fold in the human tyrosyl-tRNA synthetase. Comparison of the forward rate constants for catalysis of tyrosine activation by the human and B. stearothermophilus enzymes indicates that despite differences in their active sites and the potassium requirement of the human enzyme, the activation energies for tyrosine activation are identical for the two enzymes. The results of these investigations suggest that differences exist between the active sites of the bacterial and human tyrosyl-tRNA synthetases that could be exploited to design antimicrobials that target the bacterial enzyme. PMID- 11856732 TI - Stat3 beta inhibits gamma-globin gene expression in erythroid cells. AB - We demonstrated previously gamma-globin gene inhibition in K562 cells and primary erythroid progenitors treated with interleukin-6. Although several cis-acting elements have been identified in the globin promoters, the precise mechanism for cytokine-mediated globin gene regulation remains to be elucidated. In this report we demonstrate inhibitors of Stat3 phosphorylation abrogate interleukin-6 mediated gamma gene silencing in erythroid cells. DNA-protein binding studies established Stat3 interaction in the 5'-untranslated gamma-globin promoter region. Furthermore, co-transfection experiments with Stat3 beta demonstrate gamma promoter inhibition in a concentration-dependent manner, which was significantly reversed when the cognate Stat3-binding site in the 5'-untranslated region was mutated. These studies establish a novel mechanism for gamma gene silencing through the STAT signal transduction pathway. PMID- 11856733 TI - Chronic ethanol increases lipopolysaccharide-stimulated Egr-1 expression in RAW 264.7 macrophages: contribution to enhanced tumor necrosis factor alpha production. AB - Increased production of tumor necrosis factor alpha (TNFalpha) is associated with the development of alcoholic liver disease. Culture of RAW264.7 macrophages with 25 mm ethanol for 48 h increased lipopolysaccharide (LPS)-stimulated accumulation of tumor necrosis factor alpha (TNFalpha) peptide and mRNA by 2-fold. We investigated whether chronic ethanol-induced increases in the DNA binding and/or promoter activity of the key transcription factors regulating LPS-stimulated TNFalpha promoter activity contribute to increased TNFalpha expression. Binding of Egr-1 to the TNFalpha promoter was increased by 2.5-fold after ethanol exposure, whereas NFkappaB binding was decreased to 30% of control. AP-1 binding was not affected. Changes in binding activity were paralleled by an increased contribution of the Egr-1 binding site and a decreased contribution of the NFkappaB site to LPS-stimulated TNFalpha promoter activity. Overexpression of dominant negative Egr-1 prevented the ethanol-induced increase in LPS-stimulated TNFalpha mRNA accumulation. Chronic ethanol exposure enhanced LPS-stimulated Egr 1 promoter-driven CAT expression and transcription of Egr-1. Induction of Egr-1 is dependent on ERK1/2 activation in other systems. Therefore, we investigated whether the ERK1/2 pathway mediated the chronic ethanol-induced increases in Egr 1 and TNFalpha. Increased Egr-1 promoter activity and TNFalpha mRNA accumulation after chronic ethanol were both prevented by overexpression of dominant negative ERK1/2. LPS-stimulated ERK1/2 phosphorylation was increased 2-fold in cells cultured with ethanol compared with controls. These results demonstrate that enhanced LPS-dependent activation of Egr-1 contributes to increased TNFalpha production after chronic ethanol exposure. PMID- 11856734 TI - Anchoring of surface proteins to the cell wall of Staphylococcus aureus. III. Lipid II is an in vivo peptidoglycan substrate for sortase-catalyzed surface protein anchoring. AB - Surface proteins of Staphylococcus aureus are anchored to the cell wall peptidoglycan by a mechanism requiring a C-terminal sorting signal with an LPXTG motif. Surface proteins are first synthesized in the bacterial cytoplasm and then transported across the cytoplasmic membrane. Cleavage of the N-terminal signal peptide of the cytoplasmic surface protein P1 precursor generates the extracellular P2 species, which is the substrate for the cell wall anchoring reaction. Sortase, a membrane-anchored transpeptidase, cleaves P2 between the threonine (T) and the glycine (G) of the LPXTG motif and catalyzes the formation of an amide bond between the carboxyl group of threonine and the amino group of cell wall cross-bridges. We have used metabolic labeling of staphylococcal cultures with [(32)P]phosphoric acid to reveal a P3 intermediate. The (32)P-label of immunoprecipitated surface protein is removed by treatment with lysostaphin, a glycyl-glycine endopeptidase that separates the cell wall anchor structure. Furthermore, the appearance of P3 is prevented in the absence of sortase or by the inhibition of cell wall synthesis. (32)P-Labeled cell wall anchor species bind to nisin, an antibiotic that is known to form a complex with lipid II. Thus, it appears that the P3 intermediate represents surface protein linked to the lipid II peptidoglycan precursor. The data support a model whereby lipid II linked polypeptides are incorporated into the growing peptidoglycan via the transpeptidation and transglycosylation reactions of cell wall synthesis, generating mature cell wall-linked surface protein. PMID- 11856736 TI - Dichotomy of Ca2+ signals triggered by different phospholipid pathways in antigen stimulation of human mast cells. AB - Mast cell activation triggers Ca(2+) signals and the release of enzyme-containing granules, events that play a major role in allergic/hypersensitivity reactions. However, the precise molecular mechanisms that regulate antigen-triggered degranulation and Ca(2+) fluxes in human mast cells are still poorly understood. Here we show, for the first time, that a receptor can trigger Ca(2+) via two separate molecular mechanisms. Using an antisense approach, we show that IgE antigen stimulation of human bone marrow-derived mast cells triggers a sphingosine kinase (SPHK) 1-mediated fast and transient Ca(2+) release from intracellular stores. However, phospholipase C (PLC) gamma1 triggers a second (slower) wave of calcium release from intracellular stores, and it is this PLCgamma1-generated signal that is responsible for Ca(2+) entry. Surprisingly, FcepsilonRI (a high affinity receptor for IgE)-triggered mast cell degranulation depends on the first, sphingosine kinase-mediated Ca(2+) signal. These two pathways act independently because antisense knock down of either enzyme does not interfere with the activity of the other enzyme. Of interest, similar to PLCgamma1, SPHK1 translocates rapidly to the membrane after FcepsilonRI cross linking. Here we also show that SPHK1 activity depends on phospholipase D1 and that FcepsilonRI-triggered mast cell degranulation depends primarily on the activation of both phospholipase D1 and SPHK1. PMID- 11856737 TI - Endogenous G protein-coupled receptor kinase 6 Regulates M3 muscarinic acetylcholine receptor phosphorylation and desensitization in human SH-SY5Y neuroblastoma cells. AB - We have previously shown that overexpression of G protein-coupled receptor kinase 6 (GRK6) enhanced the phosphorylation and desensitization of the endogenously expressed M(3) muscarinic acetylcholine (mACh) receptor in human SH-SY5Y neuroblastoma cells. In this study we have examined the potential role of endogenous GRK6 in the regulation of M(3) mACh receptor by blocking its action through the introduction of a kinase-dead, dominant-negative GRK6 ((K215R)GRK6). (K215R)GRK6 expression inhibited methacholine-stimulated M(3) mACh receptor phosphorylation by 50% compared with plasmid transfected control cells. Guanosine 5'-O-(3-[(35)S]thio)triphosphate binding and immunoprecipitation studies, conducted after agonist pretreatment (3 min), indicated that M(3) mACh receptor-G alpha(q/11) uncoupling was attenuated by 50% in cells expressing (K215R)GRK6 when compared with control cells. In contrast, expression of the related dominant negative kinase (K215R)GRK5 had no effect on M(3) mACh receptor phosphorylation or uncoupling. Time course studies also showed that agonist-stimulated [(3)H]inositol phosphate accumulations were more sustained in cells expressing (K215R)GRK6 compared with control and (K215R)GRK5-expressing cells, whereas (K215R)GRK6 expression had no effect on the phospholipase C response to direct stimulation of G proteins with AlF(4)(-). The ability of (K215R)GRK6 to inhibit agonist-mediated M(3) mACh receptor phosphorylation and G protein uncoupling suggests that endogenous GRK6 mediates, at least in part, M(3) mACh receptor desensitization in the SH-SY5Y cell line. PMID- 11856739 TI - Yeast ribosomal protein L12 is a substrate of protein-arginine methyltransferase 2. AB - Type III protein-arginine methyltransferase from the yeast Saccharomyces cerevisiae (RMT2) was expressed in Escherichia coli and purified to apparent homogeneity. The cytosolic, ribosomal, and ribosome salt wash fractions from yeast cells lacking RMT2 were used as substrates for the recombinant RMT2. Using S-adenosyl-l-methionine as co-substrate, RMT2 methylated a protein in the ribosome salt wash fraction. The same protein in the ribosomal fraction was also methylated by RMT2 after pretreating the sample with endonuclease. Amino acid analysis affirmed that the labeling products were delta-N-monomethylarginines. The methylated protein from the ribosomal or the ribosome salt wash fraction was isolated by two-dimensional gel electrophoresis and identified as ribosomal protein L12 by mass spectrometry. Using synthetic peptides, recombinant L12, and its mutant as substrates, we pinpointed Arg(67) on ribosomal protein L12 as the methyl acceptor. L12 was isolated from wild type yeast cells that have been grown in the presence of S-adenosyl-l-[methyl-(3)H]methionine and subjected to amino acid analysis. The results indicate that L12 contains delta-N monomethylarginines. PMID- 11856738 TI - Suppression of androgen receptor transactivation by Pyk2 via interaction and phosphorylation of the ARA55 coregulator. AB - The proline-rich tyrosine kinase 2 (Pyk2) was first identified as a key kinase linked to the MAP kinase and JNK signaling pathways that play important roles in cell growth and adhesion. The linkage between Pyk2 and the androgen receptor (AR), an important transcription factor in prostate cancer progression, however, remains unclear. Here we report that using the full-length androgen receptor associated protein, ARA55, coregulator as bait, we were able to isolate an ARA55 interacting protein, Pyk2, and demonstrated that Pyk2 could repress AR transactivation via inactivation of ARA55. This inactivation may result from the direct phosphorylation of ARA55 by Pyk2 at tyrosine 43, impairing the coactivator activity of ARA55 and/or sequestering ARA55 to reduce its interaction with AR. Our finding that Pyk2 can indirectly modulate AR function via interaction and/or phosphorylation of ARA55 not only expands the role of Pyk2 in AR-mediated prostate cancer growth but also strengthens the role of ARA55 as an AR coregulator. PMID- 11856735 TI - Activation of constitutive nitric-oxide synthase activity is an early signaling event induced by ionizing radiation. AB - Ionizing radiation at clinical dose levels activates both pro- and anti proliferative signal transduction pathways, the balance of which determines cell fate. The initiating and amplifying mechanisms involved in the activation are poorly understood. We demonstrate that one mechanism involves stimulation of constitutive nitric-oxide synthase (NOS) activity. NOS activity of Chinese hamster ovary cells was measured by the arginine --> citrulline conversion assay. Irradiation stimulated a transient activation of NOS with maximal activity at 5 min of post-irradiation. Western blot analysis and genetic manipulation by overexpression of wild type or dominant negative NOS mutant identify the radiation-induced isoform as NOS-1. Further evidence that NOS-1 is activated by radiation was the demonstration of radiation-induced cGMP formation in cells transiently transfected with the NO-dependent soluble guanylate cyclase. Protein Tyr nitration, a footprint of peroxynitrite formation, followed radiation exposure and was inhibited by expression of a dominant negative NOS-1 mutant. Radiation-induced ERK1/2 kinase activity, a cytoprotective response to radiation, was also blocked by inhibiting NOS activity. These experiments establish NO dependent signal transduction pathways as being radio-responsive. Given the lipophilic and relatively stable properties of NO, these results also suggest a possible mechanism by which ionization events in one cell may activate signaling processes in adjacent cells. PMID- 11856740 TI - Tight binding inhibition of protein phosphatase-1 by phosphatidic acid. Specificity of inhibition by the phospholipid. AB - Phosphatidic acid (PA) has been identified as a bioactive lipid second messenger, yet despite extensive investigation, no cellular target has emerged as a mediator of its described biological effects. In this study, we identify the gamma isoform of the human protein phosphatase-1 catalytic subunit (PP1c gamma) as a high affinity in vitro target of PA. PA inhibited the enzyme dose-dependently with an IC(50) of 15 nm. Mechanistically, PA inhibited the enzyme noncompetitively with the kinetics of a tight binding inhibitor and a K(i) value of 0.97 +/- 0.24 nm. Together, these data describe one of the most potent in vitro effects of PA. To further elucidate the interaction between PA and PP1c gamma, structure/function analysis of the lipid was carried out using commercially available and synthetically generated analogs of PA. These studies disclosed that the lipid protein interaction is dependent on the presence of the lipid phosphate as well as the presence of the fatty acid side chains, because lipids lacking either of these substituents resulted in complete loss of inhibition. However, the specific composition of the fatty acid side chains was not important for inhibition. Using 1-O-hexadecyl,2-oleoyl-PA, it was also shown that the carbonyl group of the sn-1 acyl linkage is not required for the lipid-protein interaction. Finally, using a lipid-protein overlay assay, it was demonstrated that PP1c gamma specifically and directly interacts with phosphatidic acid while not significantly binding other phospholipids. These results identify PA as a tight binding and specific inhibitor of PP1, and they raise the hypothesis that PP1c gamma may function as a mediator of PA action in cells. They also argue for the existence of a specific high affinity PA-binding domain on the enzyme. PMID- 11856741 TI - Transferrin receptor-dependent iron uptake is responsible for doxorubicin mediated apoptosis in endothelial cells: role of oxidant-induced iron signaling in apoptosis. AB - In the past, investigators have successfully used iron chelators to mitigate the cardiotoxicity of doxorubicin (DOX), a widely used anticancer drug that induces reactive oxygen species (ROS), oxidative damage, and apoptosis. Although intracellular iron plays a critical role in initiating DOX-induced apoptosis, the molecular mechanism(s) that link iron, ROS, and apoptosis are still unknown. In this study, we demonstrate that apoptosis results from the exposure of bovine aortic endothelial cells to DOX and that the apoptotic cell death is accompanied by a significant increase in cellular iron ((55)Fe) uptake and activation of iron regulatory protein-1. Furthermore, DOX-induced iron uptake was shown to be mediated by the transferrin receptor (TfR)-dependent mechanism. Treatment with the anti-TfR antibody (IgA class) dramatically inhibited DOX-induced apoptosis, iron uptake, and intracellular oxidant formation as measured by fluorescence using dichlorodihydrofluorescein. Treatment with cell-permeable iron chelators and ROS scavengers inhibited DOX-induced cellular (55)Fe uptake, ROS formation, and apoptosis. Based on these findings, we conclude that DOX-induced iron signaling is regulated by the cell surface TfR expression, intracellular oxidant levels, and iron regulatory proteins. The implications of TfR-dependent iron transport in oxidant-induced apoptosis in endothelial cells are discussed. PMID- 11856742 TI - TRPC4 can be activated by G-protein-coupled receptors and provides sufficient Ca(2+) to trigger exocytosis in neuroendocrine cells. AB - Transient receptor potential (TRP) channels form a large family of plasma membrane cation channels. Mammalian members of the "short" TRP family (TRP channel (TRPC) 1-7 are Ca(2+)-permeant, non-selective cation channels that are widely expressed in various cell types, including neurons. TRPC activity is linked through unknown mechanisms to G-protein-coupled receptors or receptor tyrosine kinases that activate phospholipase C. To investigate the properties and function of TRPC4 in neuronally derived cells, we transiently expressed mouse TRPC4 and histamine H(1) receptor in mouse adrenal chromaffin cells and PC12 cells. Histamine, but not thapsigargin, stimulated Mn(2+) influx in transfected cells. In the whole-cell patch clamp mode, histamine triggered a transient current in TRPC4-expressing cells. No current was evoked by perfusion with inositol 1,4,5-trisphosphate. When exocytosis was monitored with the capacitance detection technique, the magnitude of the membrane capacitance increase (Delta C(m)) on application of histamine in H(1) receptor/TRPC4-expressing chromaffin cells was comparable with that triggered by a train of depolarizing pulses. Our results indicate that TRPC4 channels behave as receptor, but not store-operated, channels in neuronally derived cells. TRPC4 channels can provide sufficient Ca(2+) influx to trigger a robust secretory response in voltage-clamped neurosecretory cells. Similar mechanisms may modulate exocytosis in other neuronal systems. PMID- 11856744 TI - Control of actin dynamics by proteins made of beta-thymosin repeats: the actobindin family. AB - Actobindin is an actin-binding protein from amoeba, which consists of two beta thymosin repeats and has been shown to inhibit actin polymerization by sequestering G-actin and by stabilizing actin dimers. Here we show that actobindin has the same biochemical properties as the Drosophila or Caenorhabditis elegans homologous protein that consists of three beta-thymosin repeats. These proteins define a new family of actin-binding proteins. They bind G-actin in a 1:1 complex with thermodynamic and kinetic parameters similar to beta-thymosins. Like beta-thymosins, they slow down nucleotide exchange on G actin and make a ternary complex with G-actin and Latrunculin A. On the other hand, they behave as functional homologs of profilin because their complex with MgATP-G-actin, unlike beta-thymosin-actin, participates in filament barbed end growth, like profilin-actin complex. Therefore these proteins play an active role in actin-based motility processes. In addition, proteins of the actobindin family interact with the pointed end of actin filaments and inhibit pointed end growth, maybe via the interaction of the beta-thymosin repeats with two terminal subunits. PMID- 11856743 TI - Activation of the c-Jun N-terminal kinase (JNK) signaling pathway is essential during PBOX-6-induced apoptosis in chronic myelogenous leukemia (CML) cells. AB - The mitogen-activated protein (MAP) kinase family is activated in response to a wide variety of external stress signals such as UV irradiation, heat shock, and many chemotherapeutic drugs and leads to the induction of apoptosis. A novel series of pyrrolo-1,5-benzoxazepines have been shown to potently induce apoptosis in chronic myelogenous leukemia (CML) cells, which are resistant to many chemotherapeutic agents. In this study we have delineated part of the mechanism by which a representative compound known as PBOX-6 induces apoptosis. We have investigated whether PBOX-6 induces activation of MAP kinase signaling pathways in CML cells. Treatment of K562 cells with PBOX-6 resulted in the transient activation of two JNK isoforms, JNK1 and JNK2. In contrast, PBOX-6 did not activate the extracellular signal-regulated kinase (ERK) or p38. Apoptosis was found to occur independently of the small GTPases Ras, Rac, and Cdc42 but involved phosphorylation of the JNK substrates, c-Jun and ATF-2. Pretreatment of K562 cells with the JNK inhibitor, dicoumarol, abolished PBOX-6-induced phosphorylation of c-Jun and ATF-2 and inhibited the induced apoptosis, suggesting that JNK activation is an essential component of the apoptotic pathway induced by PBOX-6. Consistent with this finding, transfection of K562 cells with the JNK scaffold protein, JIP-1, inhibited JNK activity and apoptosis induced by PBOX-6. JIP-1 specifically scaffolds JNK, MKK7, and members of the mixed-lineage kinase (MLK) family, implicating these kinases upstream of JNK in the apoptotic pathway induced by PBOX-6 in K562 cells. PMID- 11856745 TI - Beta-catenin is essential and sufficient for skeletal myogenesis in P19 cells. AB - Wnt1 and Wnt3a are signaling factors known to play a role in the induction of myogenesis in the myotome of the differentiating somite. Both factors may transduce their signal by a conserved pathway that leads to transcriptional regulation by beta-catenin/Lef1. beta-Catenin and Lef1 are found in the myotome prior to MyoD expression. We have utilized the P19 cell system to study the mechanisms by which Wnt3a may activate MyoD expression and subsequent skeletal muscle development. We have isolated P19 cell lines that stably express either Wnt3a or activated beta-catenin and found that aggregation of these cells results in the induction of myogenesis compared with control cells. Pax3, Gli2, Mox1, and Six1 were expressed during Wnt3a and beta-catenin-induced differentiation prior to MyoD expression. Furthermore, we have shown that the nuclear function of beta catenin was essential for skeletal myogenesis in P19 cells by overexpression of a dominant negative beta-catenin/engrailed chimera. Primitive streak factors were present, but expression of Pax3, Mox1, Gli2, and Six1 was lost in these cells, indicating that nuclear beta-catenin is essential for specification of mesodermal precursors to the myogenic lineage. Therefore, Wnt signaling, acting via beta catenin, is necessary and sufficient for skeletal myogenesis in P19 cells. PMID- 11856747 TI - Characterization of dominant-negative mutants of the DEAH-box splicing factors Prp22 and Prp16. AB - Saccharomyces cerevisiae Prp22 and Prp16 are RNA-dependent ATPases required for pre-mRNA splicing. Both proteins are members of the DEXH-box family of nucleic acid-dependent NTPases. Prior mutational analysis of Prp22 and Prp16 identified residues within conserved motifs I (GXGKT), II (DEAH), and VI (QRXGRXGR) that are required for their biological activity. Nonfunctional Prp22 and Prp16 mutants exerted a dominant negative effect on cell growth. Here we show that overexpression of lethal Prp22 mutants leads to accumulation of unspliced pre mRNAs and excised introns in vivo. The biochemical basis for the lethality and inhibition of splicing in vivo was determined by purifying and characterizing recombinant mutant proteins. The lethal Prp22 mutants D603A and E604A in motif II and Q804A and R808A in motif VI were defective for ATP hydrolysis and mRNA release from the spliceosome, but were active in promoting step 2 transesterification. Lethal Prp16 mutants G378A and K379A in motif I; D473A and E474A in motif II; and Q685A, G688A, R689A, and R692A in motif VI were defective for ATP hydrolysis and step 2 transesterification chemistry. The ATPase-defective mutants of Prp16 and Prp22 bound to spliceosomes in vitro and blocked the function of the respective wild-type proteins in trans. Comparing the mutational effects in Prp16 and Prp22 highlights common as well as distinct structural requirements for the ATP-dependent steps in pre-mRNA splicing. PMID- 11856746 TI - Direct interaction of the NifL regulatory protein with the GlnK signal transducer enables the Azotobacter vinelandii NifL-NifA regulatory system to respond to conditions replete for nitrogen. AB - The Azotobacter vinelandii NifL-NifA regulatory system integrates metabolic signals for redox, energy, and nitrogen status to fine tune regulation of the synthesis of molybdenum nitrogenase. The NifL protein utilizes discrete mechanisms to perceive these signals leading to the formation of a protein protein complex, which inhibits NifA activity. Whereas redox signaling is mediated via a flavin-containing PAS domain in the N-terminal region of NifL, the nitrogen status is sensed via interaction with PII-like signal transduction proteins and small molecular weight effectors. The nonuridylylated form of the PII-like protein encoded by A. vinelandii glnK (Av GlnK) stimulates NifL to inhibit transcriptional activation by NifA in vitro. Here we demonstrate that the nonmodified form of Av GlnK directly interacts with A. vinelandii NifL and that this interaction is dependent on Mg(2+), ATP, and 2-oxoglutarate. Differences were observed in the regulation of the Av GlnK-NifL interaction by 2-oxoglutarate compared with the role of this effector in modulating the interaction of enteric PII-like proteins with their receptors. We also report that the interaction between Av GlnK and NifL is abolished by site-directed substitution of a single amino acid in the T-loop region of Av GlnK and that uridylylation of the conserved tyrosine residue in the T-loop inhibits the interaction. No association was detected between Av GlnK and the N-terminal region of NifL and our results demonstrate that Av GlnK directly interacts with the C-terminal histidine protein kinase-like domain. PMID- 11856750 TI - Transcription termination: primary intermediates and secondary adducts. AB - In living organisms, stable elongation complexes of RNA polymerase dissociate at specific template positions in a process of transcription termination. It has been suggested that the dissociation is not the immediate cause of termination but is preceded by catalytic inactivation of the elongation complex. In vitro reducing ionic strength can be used to stabilize very unstable and catalytically inactive complex at the point of termination; the previous biochemical characterization of this complex has led to important conclusions regarding termination mechanism. Here we analyze in detail the complexes formed between DNA template, nascent RNA, and Escherichia coli RNA polymerase during transcription through the tR2 terminator of bacteriophage lambda. At low ionic strength, the majority of elongation complexes fall apart upon reaching the terminator. Released RNA and DNA efficiently rebind RNA polymerase (RNAP) and form binary RNAP.RNA and RNAP.DNA complexes, which are indistinguishable from binary complexes obtained by direct mixing of the purified nucleic acids and the enzyme. A small fraction of elongation complexes that reach termination point escapes dissociation because RNA polymerase has backtracked from the terminator to an upstream DNA position. Thus, transcription elongation to a terminator site produces no termination intermediates that withstand dissociation in the time scale appropriate for biochemical studies. PMID- 11856749 TI - Comparison of the post-transcriptional regulation of the mRNAs for the surface proteins PSA (GP46) and MSP (GP63) of Leishmania chagasi. AB - MSP (GP63) and PSA (GP46) are abundant 63- and 46-kDa glycolipid-anchored proteins on the surface of the promastigote form of most Leishmania species. MSP is a zinc metalloprotease that confers resistance to host complement-mediated lysis. PSA contains internal repeats of 24 amino acids, and its function is unknown. The steady state levels of mRNAs for both glycoproteins are regulated post-transcriptionally, resulting in about a 30-fold increase as Leishmania chagasi promastigotes grow in vitro from logarithmic phase to stationary phase. Previous studies showed the 3'-untranslated regions (3'-UTRs) of these mRNAs are essential for this post-transcriptional regulation. These two 3'-UTRs of 1.0 and 1.3 kilobases were cloned immediately downstream of a beta-galactosidase reporter gene in a plasmid, and segments were systematically deleted to examine which portions of the 3'-UTRs contribute to the post-transcriptional regulation. The 92 nucleotide segment of greatest similarity between the two 3'-UTRs was deleted without loss of regulation, but the segments flanking this similarity region have positive regulatory elements essential for the regulation. We propose that similar, but non-identical, molecular mechanisms regulate the parallel expression of these two L. chagasi mRNAs despite their lack of sequence identity. These post transcriptional mechanisms resemble the mechanism recently suggested for the regulation of mRNAs encoding the dipeptide (EP) and pentapeptide (GPEET) repeat proteins in Trypanosoma brucei that involves interactions between positive and negative regulatory elements in the 3'-UTR. PMID- 11856751 TI - Cubilin, a binding partner for galectin-3 in the murine utero-placental complex. AB - Galectin-3 is a lectin important in animal development and regulatory processes and is found selectively localized at the implantation site of the mouse embryo. To better understand the role of galectin-3 at the maternal-fetal interface, a binding partner was isolated and characterized. Homogenates of uteroplacental tissue were incubated with immobilized recombinant galectin-3, and specifically bound proteins were eluted using lactose. The principal protein, p400, had an M(r) of 400,000 in SDS-PAGE. Physical properties of p400 and amino acid sequences of seven tryptic peptides were similar to cubilin from rats, humans, and dogs, identifying p400 as the murine ortholog of cubilin. This was further supported by the tissue distribution observed only in yolk sac, kidney, and ileum with monospecific antiserum for p400. Cubilin occurred in yolk sac epithelium throughout pregnancy, but galectin-3 was there only during the last week. Unexpectedly, cubilin was found only in perforin-containing granules of uterine natural killer (uNK) cells, although galectin-3 occurred throughout the cell cytoplasm. In situ hybridization revealed cubilin mRNA in yolk sac epithelium but not uNK cells, implying that yolk sac-derived cubilin is endocytosed by uNK cells via galectin-3. This is consistent with cubilin being an endogenous partner of galectin-3 at the maternal-fetal interface and suggests an important role for cubilin in uNK cell function. PMID- 11856748 TI - The androgen receptor can promote beta-catenin nuclear translocation independently of adenomatous polyposis coli. AB - We provide evidence that the androgen receptor (AR) can promote nuclear translocation of beta-catenin in LNCaP and PC3 prostate cancer cells. Using AR expressing cells (LNCaP) and non-AR-expressing cells (PC3) we showed by time course cell fractionation that the AR can shuttle beta-catenin into the nucleus when exposed to exogenous androgen. Cells exposed to the synthetic androgen, R1881, show distinct, punctate, nuclear co-localization of the AR and beta catenin. We further showed that the AR does not interact with adenomatous polyposis coli or glycogen synthase kinase-3beta and, therefore, conclude that androgen-mediated transport of beta-catenin occurs through a distinct pathway. The minimal necessary components of the AR and beta-catenin required for binding nuclear accumulation of beta-catenin nuclear import appears to be the DNA/ligand binding regions and the Armadillo repeats of beta-catenin. We also employed a novel DNA binding assay to illustrate that beta-catenin has the capacity to bind to the probasin promoter in an AR-dependent manner. The physiological relevance of AR-mediated transport of beta-catenin and binding to an AR promoter appeared to be a substantial increase in AR transcriptional reporter activity. AR-mediated import represents a novel mode of nuclear accumulation of beta-catenin. PMID- 11856754 TI - Enhanced apoptosis of B and T lymphocytes in TAFII105 dominant-negative transgenic mice is linked to nuclear factor-kappa B. AB - The general transcription factor TFIID is composed of the TATA-binding protein (TBP) and 12-14 TBP-associated factors (TAF(II)s). Some TAF(II)s act as bridges between transcription activators and the general transcription machinery through direct interaction with activation domains. Although TAF-mediated transcription activation has been established, there is little genetic evidence connecting it to binding of an activator. TAF(II)105 is a substoichiometric subunit of transcription factor IID highly expressed in B lymphocytes. In this study, we examined the physiological role of TAF(II)105 and its mechanism of action in vivo by expressing two forms of dominant-negative mutant TAF(II)105 in mice. We show that TAF(II)105 has a pro-survival role in B and T lymphocytes, where the native protein is expressed. In addition, TAF(II)105 is important for T cell maturation and for production of certain antibody isotypes. These phenotypic alterations were absent in mice expressing a dominant-negative mutant that lacks one of the domains mediating p65/RelA binding in vitro. These findings provide support to the notion that interaction between the activator and TAF is important for their function in vivo. PMID- 11856753 TI - Aspartic acid residues 72 and 75 and tyrosine-sulfate 73 of heparin cofactor II promote intramolecular interactions during glycosaminoglycan binding and thrombin inhibition. AB - We used site-directed mutagenesis to investigate the role of Glu(69), Asp(70), Asp(71), Asp(72), Tyr-sulfate(73), and Asp(75) in the second acidic region (AR2) of the serpin heparin cofactor II (HCII) during formation of the thrombin.HCII complex with and without glycosaminoglycans. E69Q/D70N/D71N recombinant (r)HCII, D72N/Y73F/D75N rHCII, and E69Q/D70N/D71N/D72N/Y73F/D75N rHCII were prepared to localize acidic residues important for thrombin inhibition. Interestingly, D72N/Y73F/D75N rHCII had significantly enhanced thrombin inhibition without glycosaminoglycan (4-fold greater) and with heparin (6-fold greater), showing maximal activity at 2 microg/ml heparin compared with wild-type recombinant HCII (wt-rHCII) with maximal activity at 20 microg/ml heparin. The other rHCII mutants had lesser-enhanced activities, but they all eluted from heparin-Sepharose at significantly higher ionic strengths compared with wt-rHCII. Neutralizing and reversing the charge of Asp(72), Tyr-sulfate(73), and Asp(75) were done to characterize their individual contribution to HCII activity. Only Y73K rHCII and D75K rHCII have significantly increased heparin cofactor activity compared with wt-rHCII; however, all of the individual rHCII mutants required substantially less glycosaminoglycan at maximal inhibition than did wt-rHCII. Inhibition of either alpha-thrombin/hirugen or gamma(T)-thrombin (both with an altered anion binding exosite-1) by the AR2 rHCII mutants was similar to wt-rHCII. D72N/Y73F/D75N rHCII and D75K rHCII were significantly more active than wt-rHCII in a plasma-based thrombin inhibition assay with glycosaminoglycans. These results indicate that improved thrombin inhibition in the AR2 HCII mutants is mediated by enhanced interactions between the acidic domain and anion-binding exosite-1 of thrombin and that AR2 may be a "molecular rheostat" to promote thrombin inhibition in the presence of glycosaminoglycans. PMID- 11856752 TI - The lysosomal transport of prosaposin requires the conditional interaction of its highly conserved d domain with sphingomyelin. AB - Lysosomal prosaposin (65 kDa) is a nonenzymic protein that is transported to the lysosomes in a mannose 6-phosphate-independent manner. Selective deletion of the functional domains of prosaposin indicates that the D domain and the carboxyl terminal region are necessary for its transport to the lysosomes. Inhibitors of sphingolipid biosynthesis, such as fumonisin B(1) (FB(1)) and tricyclodecan-9-yl xanthate potassium salt (D609), also interfere with the trafficking of prosaposin to lysosomes. In this study, we examine sphingomyelin as a direct candidate for the trafficking of prosaposin. Chinese hamster ovary and COS-7 cells overexpressing prosaposin or an albumin/prosaposin construct were incubated with these inhibitors, treated with sphingolipids, and then immunostained. Sphingomyelin restored the immunostaining in lysosomes in both FB(1)- and D609 treated cells and ceramide reestablished the immunostaining in FB(1)-treated cells only. D-Threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol (PDMP), which inhibits glycosphingolipids, had no effect on the immunostaining pattern. To determine whether sphingomyelin has the same effect on the transport of endogenous prosaposin, testicular explants were treated with FB(1) and D609. Sphingomyelin restored prosaposin immunogold labeling in the lysosomes of FB(1)- and D609-treated Sertoli cells, whereas ceramide restored the label in FB(1) treatment only. Albumin linked to the D and COOH-terminal domains of prosaposin was used as a dominant negative competitor. The construct blocked the targeting of prosaposin and induced accumulation of membrane in the lysosomes, demonstrating that the construct uses the same transport pathway as endogenous prosaposin. In conclusion, our results showed that sphingomyelin, the D domain, and its adjacent COOH-terminal region play a crucial role in the transport of prosaposin to lysosomes. Although the precise nature of this lipid-protein interaction is not well established, it is proposed that sphingomyelin microdomains (lipid rafts) are part of a mechanism ensuring correct intercellular trafficking of prosaposin. PMID- 11856755 TI - Disulfide bond formation promotes the cis- and trans-dimerization of the E cadherin-derived first repeat. AB - Cadherin is a cell adhesion molecule crucial for epithelial and endothelial cell monolayer integrity. The previously solved x-ray crystallographic structure of the E-CAD12 cis-dimer displayed an unpaired Cys(9), which protruded away from the Cys(9) on the other protomer. To investigate the possible biological function of Cys(9) within the first repeat (the E-cadherin-derived N-terminal repeat), E-CAD1 was overexpressed and secreted into the periplasmic space of Escherichia coli cells. Recombinant E-CAD1 produced a mixed monomer and dimer in an equilibrium fashion. The dimer was linked by a disulfide through Cys(9) pairing. Analysis by high pressure liquid chromatography and electron microscopy suggested the existence of oligomeric complexes. Mutation at Trp(2) appears to indicate that these oligomeric complexes trans-dimerize. Interestingly, mutation of Cys(9) affected not only the cis-dimerization, but also the trans-oligomerization of E CAD1. Accordingly, it is plausible that, under oxidative stress, the homophilic interactions of E-cadherin through E-CAD1 may be promoted and stabilized by this disulfide bond. PMID- 11856756 TI - Assessing the metabolic function of the MutT 8-oxodeoxyguanosine triphosphatase in Escherichia coli by nucleotide pool analysis. AB - In Escherichia coli the mutT gene is one of several that acts to minimize mutagenesis by reactive oxygen species. The bacterial MutT protein and its mammalian homolog have been shown to catalyze in vitro the hydrolysis of the oxidized deoxyguanosine nucleotide, 8-oxo-dGTP, to its corresponding monophosphate. Thus, the protein is thought to "sanitize" the nucleotide pool by ridding the cell of a nucleotide whose incorporation into DNA would be intensely mutagenic. However, because others have shown mutT mutations to be mutagenic under some conditions of anaerobic growth, and have shown 8-oxo-dGTP to be a poor DNA polymerase substrate, there is reason to question this model. We have devised an assay for 8-oxo-dGTP in bacterial extracts. Using this assay, which involves reversed-phase high-performance liquid chromatography and electrochemical detection, we have been unable to detect 8-oxo-dGTP in extracts of three different mutT mutants of E. coli, even after growth of the bacteria in the presence of hydrogen peroxide. Our estimated upper limit for 8-oxo-dGTP content of these bacteria is about 200 molecules/cell, corresponding to a concentration of about 0.34 microm. When 8-oxo-dGTP was added at 0.34 microm to an in vitro DNA replication system primed with a DNA template that permits scoring of replication errors and with the four normal dNTPs at their estimated intracellular concentrations, there was no detectable effect upon the frequency of replication errors. These findings lead us to question the conclusion that 8-oxo-dGTP is the most significant physiological substrate for the MutT protein. PMID- 11856757 TI - Direct evidence for nitric oxide production by a nitric-oxide synthase-like protein from Bacillus subtilis. AB - Nitric-oxide synthases (NOSs) are widely distributed among prokaryotes and eukaryotes and have diverse functions in physiology. Recent genome sequencing revealed NOS-like protein in bacteria, but whether these proteins generate nitric oxide is unknown. We therefore cloned, expressed, and purified a NOS-like protein from Bacillus subtilis (bsNOS) and characterized its catalytic parameters in both multiple and single turnover reactions. bsNOS was dimeric, bound l-Arg and 6R tetrahydrobiopterin with similar affinity as mammalian NOS, and generated nitrite from l-Arg when incubated with NADPH and a mammalian NOS reductase domain. Stopped-flow analysis showed that ferrous bsNOS reacted with O(2) to form a transient heme Fe(II)O(2) species in the presence of either Arg or the reaction intermediate N-hydroxy-l-arginine. In the latter case, disappearance of the Fe(II)O(2) species was kinetically and quantitatively coupled to formation of a transient heme Fe(III)NO product, which then dissociated to form ferric bsNOS. This behavior mirrors mammalian NOS enzymes and unambiguously shows that bsNOS can generate NO. NO formation required a bound tetrahydropteridine, and the kinetic effects of this cofactor were consistent with it donating an electron to the Fe(II)O(2) intermediate during the reaction. Dissociation of the heme Fe(III)NO product was much slower in bsNOS than in mammalian NOS. This constrains allowable rates of ferric heme reduction by a protein redox partner and underscores the utility of using a tetrahydropteridine electron donor in bsNOS. PMID- 11856758 TI - Role of integrin-linked kinase in leukocyte recruitment. AB - Chemokines modulate leukocyte integrin avidity to coordinate adhesion and subsequent transendothelial migration, although the sequential signaling pathways involved remain poorly characterized. Here we show that integrin-linked kinase (ILK), a 59-kDa serine-threonine protein kinase that interacts principally with beta(1) integrins, is highly expressed in human mononuclear cells and is activated by exposure of leukocytes to the chemokine monocyte chemoattractant protein-1. Biochemical inhibitor studies show that chemokine-triggered activation of ILK is downstream of phosphoinositide 3-kinase. In functional assays under physiologically relevant flow conditions, overexpression of wild-type ILK in human monocytic cells diminishes beta(1) integrin/vascular cell adhesion molecule 1-dependent firm adhesion to human endothelial cells. These data implicate ILK in the dynamic signaling events involved in the regulation of leukocyte integrin avidity for endothelial substrates. PMID- 11856761 TI - Cell-surface expression of the channel activating protease xCAP-1 is required for activation of ENaC in the Xenopus oocyte. AB - Sodium balance, extracellular fluid volume, and ultimately BP are maintained by precise regulation of the activity of the epithelial sodium channel (ENaC). Using a functional complementation assay in the Xenopus laevis oocyte expression system, a channel-activating protease (CAP-1) that increases ENaC activity two to threefold in the Xenopus oocyte expression system is here identified. External application of trypsin mimics the effect of Xenopus CAP-1 (xCAP-1) on ENaC activity, which can be blocked by aprotinin, a serine protease inhibitor, suggesting the existence of a novel extracellular pathway for controlling ENaC activity. Sequence analysis predicts that CAP-1 is a secreted and/or glycosyl phosphatidyl-inositol (GPI)-anchored protein. The aim of the present study was to determine whether cell-surface expression of xCAP-1 is required for ENaC activation. By site-directed mutagenesis of xCAP-1, the importance of the catalytic site, N-glycosylation, and the GPI anchor of xCAP-1 on ENaC activity were analyzed. Glycosylation or catalytic activity is not required for cell surface expression of xCAP-1, whereas the deletion of the GPI anchor consensus motif at the C-terminus of xCAP-1 (G305Stop) abolishes cell-surface expression and ENaC activation. G305Stop-mutated xCAP-1 is recovered as a secreted protein in the external medium. A catalytic mutant of xCAP-1 significantly decreased ENaC activation but did not fully abolish the effect of xCAP-1. The data indicate the critical role of the GPI anchor in ENaC activation and suggest that catalytic and noncatalytic mechanisms are involved. PMID- 11856759 TI - AUF1 Is a bcl-2 A + U-rich element-binding protein involved in bcl-2 mRNA destabilization during apoptosis. AB - We previously identified a conserved A + U-rich element (ARE) in the 3' untranslated region of bcl-2 mRNA. We have also recently demonstrated that the bcl-2 ARE interacts with a number of ARE-binding proteins (AUBPs) whose pattern changes during apoptosis in association with bcl-2 mRNA half-life reduction. Here we show that the AUBP AUF1 binds in vitro to bcl-2 mRNA. The results obtained in a yeast RNA three-hybrid system have demonstrated that the 1-257-amino acid portion of p37 AUF1 (conserved in all isoforms), containing the two RNA recognition motifs, also binds to the bcl-2 ARE in vivo. UVC irradiation-induced apoptosis results in an increase of AUF1. Inhibition of apoptosis by a general caspase inhibitor reduces this increase by 2-3-fold. These results indicate involvement of AUF1 in the ARE/AUBP-mediated modulation of bcl-2 mRNA decay during apoptosis. PMID- 11856760 TI - Acidosis mediates the upregulation of UT-A protein in livers from uremic rats. AB - Liver expresses a 49-kD UT-A protein whose abundance is increased by uremia. Chronic renal failure causes acidosis; therefore, the role of acidosis in increasing UT-A abundance was tested. Rats underwent 5/6 nephrectomy, and half were given bicarbonate mixed in their food. Bicarbonate administration significantly increased blood pH. Compared with sham-operated rats, UT-A protein abundance was significantly increased by 50% in livers from uremic, acidotic rats; bicarbonate administration prevented the increase in UT-A protein. To determine whether acidosis alone would increase UT-A protein in liver, rats were made acidotic, but not uremic, by feeding them HCl. HCl-feeding significantly lowered blood pH, increased urea excretion, and increased the abundance of the 49 kD liver UT-A protein by 36% compared with pair-fed nonacidotic rats. HCl-feeding significantly increased the abundance of the 117-kD UT-A1 protein in kidney inner medulla but did not change aquaporin-2 protein. Next, rats were fed urea to determine whether elevated blood urea would increase UT-A protein. However, urea feeding had no effect on UT-A in liver or kidney inner medulla. It was, therefore, concluded that acidosis, either directly or through a change in ammonium concentration, rather than other dietary components, stimulates the upregulation of UT-A protein in liver and kidney inner medulla. PMID- 11856762 TI - The MRP4/ABCC4 gene encodes a novel apical organic anion transporter in human kidney proximal tubules: putative efflux pump for urinary cAMP and cGMP. AB - The cyclic nucleotides cAMP and cGMP play key roles in cellular signaling and the extracellular regulation of fluid balance. In the kidney, cAMP is excreted across the apical proximal tubular membrane into urine, where it reduces phosphate reabsorption through a dipyridamole-sensitive mechanism that is not fully understood. It has long been known that this cAMP efflux pathway is dependent on ATP and is inhibited by probenecid. However, its identity and whether cGMP shares the same transporter have not been established. Here the expression, localization, and functional properties of human multidrug resistance protein 4 (MRP4) are reported. MRP4 is localized to the proximal tubule apical membrane of human kidney, and membrane vesicles from Sf9 cells expressing human MRP4 exhibit ATP-dependent transport of [(3)H]cAMP and [(3)H]cGMP. Both probenecid and dipyridamole are potent MRP4 inhibitors. ATP-dependent [(3)H]methotrexate and [(3)H]estradiol-17beta-D-glucuronide transport by MRP4 and interactions with the anionic conjugates S-(2,4-dinitrophenyl)-glutathione, N-acetyl-(2,4 dinitrophenyl)-cysteine, alpha-naphthyl-beta-D-glucuronide, and p-nitrophenyl beta-D-glucuronide are also demonstrated. In kidneys of rats deficient in the apical anionic conjugate efflux pump Mrp2, Mrp4 expression is maintained at the same level. It is concluded that MRP4 is a novel apical organic anion transporter and the putative efflux pump for cAMP and cGMP in human kidney proximal tubules. PMID- 11856763 TI - Posttranscriptional compensation for heterozygous disruption of the kidney specific NaK2Cl cotransporter gene. AB - Mice homozygous for a loss of function mutation of the kidney-specific NaK2Cl cotransporter, BSC1/NKCC2, do not survive. Here the effects of loss of one copy of the gene are studied. NKCC2 mRNA of NKCC2 +/- kidney was 55 +/- 6% of +/+, yet no differences were found between NKCC2 +/+ and +/- mice in BP, blood gas, electrolytes, creatinine, plasma renin concentration, urine volume and osmolality, ability to concentrate and dilute urine, and response to furosemide. When mice were challenged with 180 mM NH(4)Cl, plasma ammonia and urinary ammonia excretion were increased twofold and fivefold, respectively, but there was still no difference between the two genotypes. NKCC2 +/- mice had a near-normal level of NKCC2 protein and no clear change in the distribution of NKCC2 in the thick ascending limb (TAL) cells. In vitro microperfusion of isolated TAL showed no significant difference between the two genotypes in the basal and vasopressin stimulated capacity to reabsorb NaCl. There was no difference in the mRNA expressions of thiazide-sensitive NaCl cotransporter, epithelial Na channel (ENaC), aquaporin-2, ROMK, and NaKATPase. Halving the mRNA expression of NKCC2 does not affect BP or fluid balance because of compensatory factors that restore the protein level to near normal. One possible factor is a regulated increase in the movement of cytoplasmic protein to the luminal membrane leading to a restoration of functional transporter to an essentially wild type level. PMID- 11856764 TI - Potentiation of TNF-alpha-stimulated group IIA phospholipase A(2) expression by peroxisome proliferator-activated receptor alpha activators in rat mesangial cells. AB - Natural activators of peroxisome proliferator-activated receptors (PPAR) are lipid metabolites, including those produced by phospholipases A(2) (PLA(2)). In glomerular mesangial cells, the secreted group IIA PLA(2) (sPLA(2)-IIA), which is thought to be a crucial factor in pathologic processes in the kidney, may provide free fatty acids and eicosanoids directly or indirectly, by activating a cytosolic PLA(2). The scope of this study was to investigate whether synthetic PPAR(alpha) activators have an effect on sPLA(2)-IIA mRNA expression in rat mesangial cells, thus constituting a feedback modulation of sPLA(2)-IIA transcription. In the presence of tumor necrosis factor-alpha (TNF-alpha), the PPAR(alpha) agonists WY14643 and LY171883 as well as the lipid-lowering compound clofibrate potentiated expression, secretion, and activity of group IIA sPLA(2) in mesangial cells. MK886, known as a noncompetitive inhibitor of PPAR(alpha), completely abolished the potentiation of sPLA(2)-IIA secretion and activity by WY14643, thus indicating that the effect of WY14643 is specifically mediated by PPAR(alpha). When cells were transfected with different constructs of the rat sPLA(2)-IIA promoter fused to a luciferase reporter gene, a stimulation with TNF alpha in the presence of the PPAR(alpha) activators caused an enhanced promoter activity compared with that induced by TNF-alpha alone. Site-directed mutagenesis of a putative PPRE site in the sPLA(2)-IIA promoter abolished the potentiating effect of PPAR(alpha) agonists, thus strongly indicating its contribution to the enhanced promoter activity. In summary, this study shows that the rat sPLA(2)-IIA promoter is sensitive to PPAR(alpha) agonists, which act synergistically with cytokines, resulting in an enhanced expression of sPLA(2)-IIA in rat mesangial cells. PMID- 11856766 TI - A conditionally immortalized human podocyte cell line demonstrating nephrin and podocin expression. AB - Recent molecular insights have established the podocyte as a key component of the glomerular filtration barrier, and hence an important common pathway in proteinuric diseases. A conditionally immortalized human podocyte cell line has been developed by transfection with the temperature-sensitive SV40-T gene. These cells proliferate at the "permissive" temperature (33 degrees C). After transfer to the "nonpermissive" temperature (37 degrees C), they entered growth arrest and expressed markers of differentiated in vivo podocytes, including the novel podocyte proteins, nephrin, podocin, CD2AP, and synaptopodin, and known molecules of the slit diaphragm ZO-1, alpha-, beta-, and gamma-catenin and P-cadherin. The differentiation was accompanied by a growth arrest and the upregulation of cyclin dependent kinase inhibitors, p27 and p57, as well as cyclin D(1), whereas cyclin A was downregulated. These data are consistent with cell cycle protein expression during podocyte maturation in vivo. In conclusion, the development of this cell line provides a new tool in the study of podocyte biology, which will enable accurate assessment of the behavior of these complex cells in health and disease. PMID- 11856768 TI - Macrophage-stimulating protein is produced by tubular cells and activates mesangial cells. AB - Until now, hepatocytes have been the only known cell source of macrophage stimulating protein (MSP), and tissue macrophages have been the cells on which the biologic effects of MSP have been proved. To extend the understanding of the biologic meaning of MSP, it was investigated whether MSP operates in the kidney. MSP protein was evaluated by Western blot in supernatant of cultured human tubular cells (HK2) and human mesangial cells (HMC). MSP mRNA was investigated in HK2 by reverse transcription-polymerase chain reaction (RT-PCR). The expression of the MSP receptor, RON, was evaluated in HMC and HK2 by Western blot. RON mRNA was investigated in HMC by RT-PCR. The expression of MSP and RON in normal human renal tissue was studied by immunohistochemistry. HMC were stimulated with recombinant MSP (rMSP) and HK2 supernatant to study cell growth, migration, and the capacity to invade an artificial collagen matrix and synthesize interleukin-6 (IL-6). HK2 produced MSP and expressed RON in a form that was phosphorylated by rMSP. HMC expressed RON but did not produce MSP. MSP in HK2 supernatant and rMSP induced in HMC phosphorylation of RON, growth, migration, invasion, and IL-6 synthesis. In normal human kidney, tubules expressed MSP and RON. These results indicate a novel field of operation for MSP and suggest a pathogenic role of the MSP/RON system in renal disease. In fact, MSP released by tubular cells may recruit monocytes/macrophages in inflammatory tubulointerstitial disorders. In addition, MSP either circulating or as paracrine product may sustain glomerular mesangioproliferative disease. PMID- 11856770 TI - Midkine is involved in kidney development and in its regulation by retinoids. AB - In the kidney, in which development depends on epithelial-mesenchymal interactions, it has been shown that retinoids modulate nephrogenesis in a dose dependent manner in vivo and in vitro. Midkine (MK) is a retinoic acid responsive gene for a heparin-binding growth factor. The aim of the present study was therefore to quantify the expression of MK mRNA during renal development in the rat, to analyze the regulation of MK expression by retinoids in vivo and in vitro, and, finally, to study the role of MK in rat metanephric organ cultures. The spatiotemporal expression of MK in fetal kidney was studied. In control rats, MK expression is ubiquitous at gestational day 14, i.e., at the onset of nephrogenesis. On day 16, MK is expressed in the condensed mesenchyme and in early epithelialized mesenchymal derivatives. On gestational day 21, MK is rather localized in the nonmature glomeruli of the renal cortex. In utero exposure to vitamin A deficiency did not modify the specific spatial and temporal expression pattern of MK gene in the metanephros, although a decrease in mRNA expression occurred. In metanephroi explanted from 14-d-old fetuses and cultured in a defined medium, expression of MK mRNA was found to be stimulated when retinoic acid (100 nM) was added in the culture medium. Finally, in vitro nephrogenesis was strongly inhibited in the presence of neutralizing antibodies for MK: the number of nephrons formed in vitro was reduced by approximately 50% without changes in ureteric bud branching morphogenesis. These results indicated that MK is implicated in the regulation of kidney development by retinoids. These results also suggested that MK plays an important role in the molecular cascade of the epithelial conversion of the metanephric blastema. PMID- 11856767 TI - Type 1 parathyroid hormone receptor expression level modulates renal tone and plasma renin activity in spontaneously hypertensive rat. AB - These studies examine whether PTHrP(1-36), a vasodilator, modulates BP and renal vascular resistance (RVR) in spontaneously hypertensive rat (SHR). Within the kidney of normotensive rats, PTHrP(1-36) was enriched in vessels. In vessels of SHR, PTHrP was upregulated by 40% and type 1 PTH receptor (PTH1R) was downregulated by 65% compared with normotensive rats. To investigate the role of endogenous PTHrP in the regulation of BP and RVR, SHR were subjected to somatic human (h)PTH1R gene delivery. Three weeks after a single intravenous injection of pcDNA1.1 plasmid containing the hPTH1R gene under the control of the cytomegalovirus promoter, hPTH1R mRNA was detected in all of the main organs. Within the kidney, the transgene was enriched in vessels. In the isolated perfused kidney, RVR was reduced by 23% and PTHrP(1-36)-induced vasodilation, which is depressed in SHR, was restored and a vasoconstrictory response to PTH(3 34), a PTH1R antagonist, was revealed. These effects were not observed in control SHR treated with empty plasmid. BP remained unchanged, and plasma renin activity increased by 60%. Thus, in SHR renal vessels, a reduced number of PTH1R contributes to the high RVR, despite the higher expression of vasodilatory PTHrP. Moreover, these studies provide evidence for a direct link between the density of PTH1R and plasma renin activity, which might be responsible for the absence of effect of PTH1R gene delivery on BP in SHR. Overall, PTHrP significantly contributes to the homeostasis of renal and systemic hemodynamics in SHR. PMID- 11856771 TI - The p53Pro72Arg polymorphism is associated with albuminuria among aboriginal Australians. AB - Albuminuria is a widely recognized marker of renal disease and cardiovascular risk. This is especially true in Aboriginal Australians living in remote communities who suffer high rates of end-stage renal disease and cardiovascular mortality. During a survey of risk factors for renal and cardiovascular disease in one such community, an association between a common polymorphism at codon 72 (Arg/Pro) of the p53 gene and markers of renal disease was sought. A cross sectional community survey including 217 people was performed. Genotypes of the polymorphism were distributed in Hardy-Weinberg equilibrium, with p53Arg allele frequency of 0.45 (range, 0.41 to 0.50). Overall prevalence of albuminuria was high (31% microalbuminuria; 14% overt albuminuria). Urine albumin/creatinine ratio (ACR) was significantly associated with the number of p53Pro alleles (P = 0.01), and there was an interaction with tobacco smoking (P = 0.04). The p53 genotype was also associated with increasing HbA1c, but the relationship between p53 and ACR was independent of this. This is a previously unreported association. This study does not address the mechanism, but this finding, if confirmed, expands the described effects of p53 in cellular proliferation and apoptosis to include a role in the course of renal and possibly cardiovascular disease in this population. PMID- 11856765 TI - Synthesis of 1,25-dihydroxyvitamin D(3) by human endothelial cells is regulated by inflammatory cytokines: a novel autocrine determinant of vascular cell adhesion. AB - In addition to its calciotropic function, the secosteroid 1,25-dihydroxyvitamin D(3) (1,25(OH)(2)D(3)) has potent nonclassical effects. In particular, local production of 1,25D(3) catalyzed by the enzyme 1alpha-hydroxylase (1alpha-OHase) may act as an autocrine/paracrine immunomodulatory mechanism. To investigate the significance of this in vascular tissue the expression and function of 1alpha OHase in human endothelial cells was characterized. Immunohistochemical and in situ hybridization analyses show, for the first time, the presence of 1alpha OHase mRNA and protein in endothelial cells from human renal arteries as well as postcapillary venules from lymphoid tissue. Reverse transcription-PCR and Western blot analyses confirmed the presence of 1alpha-OHase in primary cultures of human umbilical vein endothelial cells (HUVEC). Enzyme activity in HUVEC (318 +/- 56 fmoles 1,25(OH)(2)D(3)/hr/mg protein) increased after treatment with tumor necrosis factor-alpha (1054 +/- 166, P < 0.01), lipopolysaccharide (1381 +/- 88, P < 0.01), or forskolin (554 +/- 56, P < 0.05). Functional studies showed that exogenously added 1,25(OH)(2)D(3) or its precursor, 25-hydroxyvitamin D(3) (25(OH)D(3)), significantly decreased HUVEC proliferation after 72 h of treatment (33% and 11%, respectively). In addition, after 24 h treatment, both 1,25(OH)(2)D(3) and 25(OH)D(3) increased the adhesion of monocytic U937 cells to HUVEC (159% and 153%, respectively). These data indicate that human endothelia are able to produce active vitamin D. The rapid induction of endothelial 1alpha OHase activity by inflammatory cytokines suggests a novel autocrine/paracrine role for the enzyme, possibly as a modulator of endothelial cell adhesion. PMID- 11856775 TI - Effects of L-carnitine supplementation in maintenance hemodialysis patients: a systematic review. AB - There are many causes for carnitine depletion during maintenance hemodialysis. Supplementation with L-carnitine in animals has been associated with improvement in some abnormalities also present in chronic renal failure. However, it is still controversial whether restoring plasma or tissue carnitine will correct clinical or biologic symptoms observed in maintenance hemodialysis. A systematic review is here performed to determine the effects of L-carnitine in maintenance hemodialysis patients. Eighty-three prospective trials were identified from 1978 to 1999 in which L-carnitine was randomly allocated in 21 trials. Change in serum triglycerides, cholesterol fractions, hemoglobin levels, erythropoietin dose, and other symptoms (muscle function, exercise capacity, and quality of life) were examined. A total of 482 patients in 18 trials were considered for analysis. There was no effect of L-carnitine on triglycerides, total cholesterol, or any of its fractions. Before the erythropoietin (EPO) era, L-carnitine treatment was associated with improved hemoglobin (P < 0.01) and with a decreased EPO dose (P < 0.01) and improved resistance to EPO when patients routinely received EPO. Muscle function, exercise capacity, and quality of life could not be reliably assessed because of the noncombinable nature of end points and the limited number of trials. In conclusion, L-carnitine cannot be recommended for treating the dyslipidemia of maintenance hemodialysis patients. By contrast, this review suggests a promising effect of L-carnitine on anemia management. The route of L carnitine administration should be evaluated because there is no evidence as to the most efficient method of administration in maintenance hemodialysis. PMID- 11856772 TI - Effects of low-dose and early versus late perindopril treatment on the progression of severe diabetic nephropathy in (mREN-2)27 rats. AB - It was previously reported that transgenic (mRen-2)27 rats with streptozotocin induced diabetes mellitus progressively develop advanced nephropathy in 12 wk. These lesions are largely prevented when the angiotensin-converting enzyme inhibitor perindopril is administered from the time of induction of diabetes mellitus. This study aimed to determine the lowest dose of early perindopril treatment required for substantial improvement of renal function and structure and to investigate whether late intervention prevents or reverses the progression of established renal lesions. At 6 wk of age, female heterozygous Ren-2 rats were randomized to receive either streptozotocin (diabetic) or citrate buffer (control). Rats were gavaged, beginning early after the induction of diabetes mellitus or the administration of control vehicle, with 0, 0.02, 0.2, or 2 mg/kg per d perindopril for 12 wk. A separate group of diabetic Ren-2 rats received late treatment with 2 mg/kg per d perindopril throughout week 8 to week 12, when rats were hypertensive and albuminuric and exhibited increased kidney weight and glomerulosclerotic index (GSI). Among diabetic rats, early 0.02 mg/kg per d perindopril treatment reduced systolic BP, GSI, and renal collagen staining but had no effect on albuminuria or kidney hypertrophy. Early 0.2 or 2 mg/kg per d perindopril treatment further reduced systolic BP, GSI, and renal collagen staining and decreased albuminuria and kidney hypertrophy. Late intervention was as antihypertensive and antialbuminuric as early 0.2 or 2 mg/kg per d perindopril treatment but did not prevent a moderate increase in GSI. In conclusion, early treatment with 0.2 mg/kg per d perindopril was the lowest dosage to largely prevent severe diabetic nephropathy in transgenic Ren-2 rats. Late-onset perindopril treatment of diabetic rats with established nephropathy was as efficacious as early treatment with respect to various renal parameters, such as albuminuria, but was associated with moderate progression of glomerulosclerosis. PMID- 11856769 TI - The effects of platelet-derived growth factor antagonism in experimental glomerulonephritis are independent of the transforming growth factor-beta system. AB - Platelet-derived growth factor B-chain (PDGF-B)- and transforming growth factor beta (TGF-beta)-mediated accumulation of extracellular matrix proteins contributes to many progressive renal diseases. In vivo, specific antagonism of either PDGF-B or TGF-beta in experimental mesangioproliferative glomerulonephritis resulted in an almost complete inhibition of matrix protein accumulation, which suggests an interaction between signaling pathways of these two growth factors. Because nothing is known on the nature of this possible interaction, PDGF-B was antagonized in the rat anti-Thy 1.1 model of glomerulonephritis by use of specific aptamers and its effects on the TGF-beta system were investigated. Antagonism of PDGF-B led to a significant reduction of glomerular matrix accumulation compared with scrambled aptamer-treated nephritic controls. PDGF-B antagonism had no effect on the overexpression of glomerular TGF beta mRNA, TGF-beta protein, or the expression of TGF-beta receptor type I and II mRNA. By immunohistology, it was possible to detect overexpression of the cytoplasmic TGF-beta signaling molecules Smad2 (agonistic) and Smad7 (antagonistic) in glomeruli of nephritic control rats which peaked on day 7 after disease induction, i.e., the peak of mesangial cell proliferation in this model. However, immunohistology and Western blot analysis again revealed no difference in the glomerular expression of both Smad proteins between PDGF-B antagonized and nonantagonized nephritic animals. In addition, no difference in the glomerular expression of phosphorylated Smad2 (P-Smad2) was detected between the differently treated nephritic groups. These observations suggest that the effects of PDGF-B antagonism are independent of TGF-beta in mesangioproliferative glomerulonephritides. PMID- 11856776 TI - Short- and long-term functional effects of percutaneous transluminal angioplasty in hemodialysis vascular access. AB - The efficacy of percutaneous transluminal angioplasty (PTA) is usually expressed as the angiographic result. Access flow (Qa) measurements offer a means to quantify the functional effects. This study was performed to evaluate the short term functional and angiographic effects of PTA and to determine the longevity of the functional effects during the follow-up period. Patients with an arteriovenous graft (AVG) or an arteriovenous fistula (AVF) who were eligible for PTA (Qa values of <600 ml/min) were included. Ultrasound-dilution Qa measurements were obtained shortly before PTA and periodically after PTA, beginning 1 wk after the procedure. The short-term effects were expressed as the increase in Qa and the reduction of stenosis. The long-term effects were expressed as patency and the decrease in Qa after PTA. Ninety-eight PTA procedures for 60 patients (65 AVG and 33 AVF) were analyzed. Qa improved from 371 +/- 17 to 674 +/- 30 ml/min for AVG and from 304 +/- 24 to 638 +/- 51 ml/min for AVF (both P < 0.0001). In 66% (AVG) and 50% (AVF) of cases, Qa increased to levels of >600 ml/min. The degree of stenosis decreased from 65 +/- 3 to 17 +/- 2% for AVG and from 72 +/- 5 to 23 +/- 7% for AVF (both P < 0.005). The reduction of stenosis was not correlated with DeltaQa (r(2) = 0.066). Six-month unassisted patency rates after PTA were 25% for AVG and 50% for AVF. The decreases in Qa were 3.7 +/- 0.8 ml/min per d for AVG and 1.8 +/- 0.9 ml/min per d for AVF. Qa values before PTA and DeltaQa were correlated with the subsequent decrease in Qa (P < 0.005). In conclusion, Qa increases after PTA but, in a substantial percentage of cases, not to levels of >600 ml/min. Qa values before PTA and the increase in Qa were correlated with long-term outcomes, whereas angiographic results were not. These data, combined with literature data, suggest that there is optimal timing for PTA. PMID- 11856773 TI - Regulation of arachidonic acid production by intracellular calcium in parathyroid cells: effect of extracellular phosphate. AB - The action of extracellular calcium on the calcium receptor in parathyroid cells results in activation of phospholipase C (PLC), PLD, and PLA(2). The PLA(2) arachidonic acid (AA) intracellular signaling pathway mediates inhibition of parathyroid hormone (PTH) secretion. In addition, stimulation of the calcium receptor produces increases in intracellular calcium levels. It was demonstrated that high extracellular phosphate levels reduce the production of AA, a mechanism by which phosphate may stimulate PTH secretion. The objective was to determine, in parathyroid tissue, whether AA production is stimulated by increases in intracellular calcium levels and to investigate whether the decreased AA production induced by high extracellular phosphate levels could be modified by increases in intracellular calcium levels. Experiments were performed in vitro using parathyroid tissue. The intracellular calcium level was increased by incubation with an ionophore (A23187), which increases calcium influx across the cell membrane, or thapsigargin, which releases calcium from intracellular stores. The phosphate concentration in the medium was normal (1 mM) or high (4 mM). The response to calcium was evaluated by incubation with 0.6 or 1.35 mM calcium concentrations. AA production by parathyroid tissue was measured by gas chromatography. In parathyroid tissue incubated with either a calcium ionophore or thapsigargin, there was an increase in AA production, together with inhibition of PTH secretion, suggesting that PLA(2) is activated by the elevation in intracellular calcium levels. Therefore, the effect of intracellular calcium level elevation on AA production in the presence of high extracellular phosphate levels was evaluated. The results demonstrate that, despite high phosphate levels in the medium, both the ionophore and thapsigargin were capable of inducing a marked increase in AA production, which was associated with a decrease in PTH secretion. In conclusion, in parathyroid tissue, AA levels can be regulated by an ionophore and thapsigargin, both of which increase cytosolic calcium concentrations. The stimulation of PTH secretion by high phosphate levels can be prevented by increases in intracellular calcium levels. PMID- 11856774 TI - Basic fibroblast growth factor among children with diarrhea-associated hemolytic uremic syndrome. AB - Diarrhea-associated hemolytic uremic syndrome (D+HUS) is characterized by endothelial injury and activation of inflammatory cytokines. Basic fibroblast growth factor (bFGF) is an angiogenic peptide released in response to vascular damage. The plasma concentrations and urinary excretion of bFGF during the course of D+HUS were determined, in comparison with the levels of various inflammatory cytokines, and changes were correlated with clinical and laboratory features of the disease. Serial plasma and urine samples were collected from 31 children with D+HUS, during the acute (days 1 to 7 of hospitalization) and recovery (through day 60 after discharge from the hospital) phases of the disease. The patients were enrolled in the multicenter trial of SYNSORB Pk (SYNSORB Biotech, Calgary, Alberta, Canada) treatment for D+HUS. bFGF, interleukin-1alpha (IL-1alpha), IL-8, and tumor necrosis factor-alpha levels were determined with enzyme-linked immunosorbent assays. bFGF was detected in urine and plasma samples more frequently than were IL-1alpha, IL-8, and tumor necrosis factor-alpha. There was an acute increase in urinary bFGF excretion, which returned to normal during convalescence. Urinary excretion of bFGF during the acute phase was higher among patients who required dialysis, compared with those who did not (48.9 +/- 15.0 and 28.9 +/- 9.0 pg/ml, respectively; P < 0.05). Plasma bFGF concentrations were persistently elevated throughout the period of hospitalization and the follow-up period among patients with D+HUS. Urinary excretion and plasma levels of bFGF were comparable for the SYNSORB Pk-treated (n = 19) and placebo-treated (n = 12) groups. Measurements of urinary and plasma concentrations of bFGF among patients with D+HUS may be useful indices for assessment of the severity of acute renal disease and the timing and adequacy of the systemic angiogenic process during early convalescence. PMID- 11856778 TI - Nocturnal hypoxemia predicts incident cardiovascular complications in dialysis patients. AB - Nocturnal hypoxemia secondary to sleep apnea has long been implicated as a cardiovascular risk factor in renal failure, but to date there is no study that links nocturnal hypoxemia to cardiovascular outcomes in end-stage renal disease. Fifty uremic patients on regular dialysis treatment without primary sleep apnea, pulmonary diseases, or other illnesses that may cause sleep apnea underwent pulse oximetry studies during night and were followed up for 32 mo. Average nocturnal SaO(2), minimal SaO(2), and the number of episodes of hypoxemia were similar in patients who died during the follow-up and in patients who survived, and none of these parameters predicted all-cause mortality. Average nocturnal SaO(2) was significantly lower (P = 0.006) in patients who had cardiovascular events during the follow-up (94.7 +/- 2.9%) than in event-free patients (97.1 +/- 1.3%). In a Cox model, average nocturnal SaO(2) was the second factor in rank explaining these outcomes. In this model a 1% decrease in average nocturnal SaO(2) was associated with a 33% increase in the incident risk of fatal and nonfatal cardiovascular events. Furthermore the risk of cardiovascular events was 5.05 times higher in patients with average nocturnal SaO(2) <95% (95% CI 1.61 to 15.86) than in those above this threshold (P = 0.005). This study adds weight to the hypothesis that nocturnal hypoxemia in dialysis patients represents an important cardiovascular risk factor. PMID- 11856777 TI - Antiangiogenic and antifibrotic gene therapy in a chronic infusion model of peritoneal dialysis in rats. AB - To identify the relative importance of peritoneal fibrosis and angiogenesis in peritoneal membrane dysfunction, adenoviral mediated gene transfer of angiostatin, a recognized angiogenesis inhibitor, and decorin, a transforming growth factor-beta-inhibiting proteoglycan, were used in a daily infusion model of peritoneal dialysis. A peritoneal catheter and subcutaneous port were inserted in rats. Five and fourteen d after insertion, adenovirus-expressing angiostatin, decorin, or AdDL70, a null control virus, were administered. Daily infusion of 4.25% Baxter Dianeal was initiated 7 d after catheter insertion and continued until day 35. Three initial doses of lipopolysaccharide were administered on days 8, 10, and 12 to promote an inflammatory response. Net ultrafiltration was used as a measure of membrane function, and peritoneum-associated vasculature and mesenteric collagen content was quantified. Ultrafiltration dysfunction, angiogenesis, and fibrosis were observed in daily infusion control animals. Animals treated with AdAngiostatin demonstrated an improvement in net ultrafiltration (-3.1 versus -7.8 ml for control animals; P = 0.0004) with a significant reduction in vessel density. AdDecorin-treated animals showed a reduction in mesenteric collagen content (1.8 versus 2.9 microg/mg; P = 0.04); however, AdDecorin treatment had no effect on net ultrafiltration. In a rodent model of peritoneal membrane failure, net ultrafiltration was significantly improved and peritoneal-associated blood vessels were significantly reduced by using adenovirus-mediated gene transfer of angiostatin. Decorin, a transforming growth factor-beta-inhibiting proteoglycan, reduced collagen content but did not affect net ultrafiltration. Improvement in the function of the peritoneum as a dialysis membrane after treatment with angiostatin has implications for treatment of peritoneal membrane dysfunction seen in patients on long-term dialysis. PMID- 11856779 TI - Iron administration and clinical outcomes in hemodialysis patients. AB - To evaluate the impact of parenteral iron administration on the survival and rate of hospitalization of US hemodialysis patients, a nonconcurrent cohort study of 10,169 hemodialysis patients in the United States in 1994 was conducted. The main outcome measures were patient survival and rate of hospitalization. After adjusting for 23 demographic and comorbidity characteristics among 5833 patients included in multivariable analysis, bills for 10 vials showed a statistically significant elevated rate of death (adjusted RR = 1.11; 95% CI, 1.00 to 1.24; P = 0.05). Bills for 10 vials showed statistically significant elevated risk (adjusted RR = 1.12; 95% CI, 1.01 to 1.25; P = 0.03). Prescribing iron in quantities of 10 vials (1000 mg) of iron dextran over a period of 6 mo. PMID- 11856780 TI - Renal insufficiency and subsequent death resulting from cardiovascular disease in the United States. AB - Several epidemiologic studies reported that persons with renal insufficiency might have increased cardiovascular disease-related mortality rates in select populations. The association between renal insufficiency and increased cardiovascular disease-related and all-cause mortality rates during 16 yr of follow-up monitoring was examined among participants who were 30 to 74 yr of age at the baseline examinations in 1976 to 1980, with urinary protein dipstick measurements (n = 8786) or serum creatinine levels of or=300 mg/dl and were 4.1, 8.6, and 20.5 deaths/1000 person-yr among participants with estimated GFR of >or=90, 70 to 89, and <70 ml/min, respectively. After adjustment for potential confounders, the relative hazards (and 95% confidence intervals) for cardiovascular disease-related death were 1.57 (0.99 to 2.48) and 1.77 (0.97 to 3.21) among subjects with urinary protein levels of 30 to 299 and >or=300 mg/dl, respectively, compared with <30 mg/dl (P trend = 0.02). The corresponding relative hazards for all-cause-related death were 1.64 (1.23 to 2.18) and 2.00 (1.13 to 3.55; P trend < 0.001). Compared with subjects with estimated GFR of >or=90 ml/min, those with estimated GFR of <70 ml/min exhibited higher relative risks of death from cardiovascular disease and all causes [1.68 (1.33 to 2.13) and 1.51 (1.19 to 1.91), respectively]. This study indicates that, in a representative sample of the United States general population, renal insufficiency is independently associated with increased cardiovascular disease related and all-cause mortality rates. PMID- 11856781 TI - Chemokine receptor polymorphism and risk of acute rejection in human renal transplantation. AB - Chemokines regulate the trafficking of leukocytes in immunity and inflammation and have been implicated in mouse models in acute cardiac and renal allograft rejection; however, their significance to human transplantation is not yet defined. The association of human chemokine receptor genetic variants, CCR5 Delta32, CCR5-59029-A/G, CCR2-V64I, CX3CR1-V249I, and CX3CR1-T280M, with outcome in 163 renal transplant recipients was examined here. Significant reductions were found in risk of acute renal transplant rejection in recipients who possessed the CCR2-64I allele (odds ratio [OR], 0.30; 95% confidence interval [CI], 0.12 to 0.78; P = 0.014) or who were homozygous for the 59029-A allele (OR, 0.37; 95% CI, 0.16 to 0.85; P = 0.016). There were no significant differences in the incidence of rejection among patients stratified as with or without CCR5-Delta32 or by the CX3CR1-V249I or CX3CR1-T280M genotypes. Adjustment for known risk factors for transplant rejection confirmed the univariate findings for possession of the CCR2 64I allele (OR, 0.20; P = 0.032) and homozygosity for the 59029-A allele (OR, 0.26; P = 0.027). It was concluded that the risk of acute rejection in renal transplantation is associated with genetic variation in the chemokine receptors CCR2 and CCR5. PMID- 11856782 TI - The pharmacokinetic-pharmacodynamic relationship for total and free mycophenolic Acid in pediatric renal transplant recipients: a report of the german study group on mycophenolate mofetil therapy. AB - It is currently being debated whether pharmacokinetic monitoring of mycophenolic acid (MPA), the active constituent of mycophenolate mofetil (MMF), can optimize MMF therapy after organ transplantation. This open-label longitudinal study in pediatric renal transplant recipients was designed to investigate the pharmacokinetic (PK)/pharmacodynamic relationship of total and free MPA and to establish PK values for the assessment of an individual's MPA PK parameters. Fifty-four children, aged 2.2 to 17.8 yr, on an immunosuppressive triple regimen consisting of cyclosporin A, prednisone, and MMF (600 mg/m(2) body surface area twice daily) were investigated 1 wk and 3 wk (initial phase) and 3 mo and 6 mo (stable phase) after renal transplantation. MPA was measured by reverse phase HPLC, free MPA by HPLC after separation by ultrafiltration. There was an association between the risk of acute rejection episodes and MPA-AUC(0-12) values or MPA predose levels; by receiver operating characteristic analysis, an AUC(0 12) of 33.8 mg x h/L in the initial phase posttransplant had a diagnostic sensitivity of 75% and a diagnostic specificity of 64% for discrimination of patients with acute rejections. The respective discrimination threshold for the MPA predose concentration was 1.2 mg/L with a sensitivity of 83% and a specificity of 64%. In contrast, high free, but not total, MPA-AUC(0-12) values were associated with an increased risk of the MMF-related side effects leukopenia and/or infections. These data indicate that therapeutic drug monitoring of MPA has the potential for optimization of MMF efficacy in this patient population by steering patients away from the low values of MPA PK variables that are associated with an increased rejection risk. For the assessment of the toxic risk of MMF regarding leukopenia and/or infections, measurement of free MPA appears to be more appropriate. PMID- 11856783 TI - Association of antibody induction with short- and long-term cause-specific mortality in renal transplant recipients. AB - A total of 73,707 primary renal transplants reported to the USRDS between 1988 and 1997 were examined to investigate the cause-specific risk for patient death associated with anti-lymphocyte antibody induction therapy (ABI). Cox proportional hazard models were used to estimate the relative risk of the use of ABI and patient death. All Cox models were corrected for potential confounding variables, such as age, gender, race, HLA mismatch, panel reactive antibody, delayed graft function, cold ischemia time, time since start of dialysis, etiology of end-stage renal disease, cytomegalovirus risk group, donor source (living or cadaveric), era effect, and immunosuppressive therapy. Primary study end points were patient death with functioning graft (DWFG) and overall patient death, including death after graft loss. Early patient death (deaths within the first 6 mo after renal transplantation) and late death (deaths after 6 mo post renal transplantation) were investigated separately. Additionally, specific causes of death were investigated. ABI was associated with a significant risk for late death after renal transplantation (relative risk [RR] = 1.1; P < 0.001) but not for DWFG (RR = 0.94; P = 0.10). ABI conferred the highest RR for late malignancy-related death (RR = 1.35; P < 0.001). ABI was significantly associated with early deaths due to infection and cardiovascular causes (RR = 1.32 [P < 0.001] and RR = 1.27 [P < 0.001], respectively). Kaplan Meier plots confirmed that the risk of ABI for patient death secondary to infectious complications was increased predominately early after transplantation as opposed to late for malignancy-related death. ABI was associated with a significant relative risk for patient death secondary to cardiovascular causes and infectious complications early in the posttransplant period. In addition, ABI was associated with a significant risk for long-term malignancy-related death. The risk of ABI should be taken in context with potential benefits of this therapy. PMID- 11856784 TI - Determinants of hypofiltration during acute renal allograft rejection. AB - This study sought to determine the extent to which GFR is decreased during acute renal allograft rejection in human subjects and to determine the mechanism of the decrease in GFR. Eight patients with biopsy-proven acute rejection were compared with 18 recipients of optimally functioning renal allografts. GFR and renal plasma flow (RPF) were measured as the clearance of inulin and para-aminohippuric acid, respectively. Arterial BP was determined, blood was sampled, and plasma oncotic pressure (pi(A)) was measured. Glomeruli obtained by biopsy during rejection were subjected to morphometric analysis, for determination of K(f). Control morphometric values for healthy glomeruli were provided by 10 living donors from whom biopsies were obtained at the time of organ donation. The subjects in the acute rejection group exhibited a significantly reduced GFR of 17 +/- 4 ml/min per 1.73 m(2), compared with 72 +/- 4 ml/min per 1.73 m(2) for control subjects (P < 0.001). With the use of a sensitivity analysis to take into account the unknown para-aminohippuric acid extraction ratio, the RPF rate was calculated to have likely been significantly decreased, by 45 to 70%, in the acute rejection group. Neither the plasma oncotic pressure nor the mean arterial pressure differed between the two groups. Morphometric analysis revealed no difference in the single-nephron K(f) values for the acute rejection group, compared with the control group. These results indicate that acute renal allograft rejection causes a profound decrease in GFR, which is attributable to a decrease in RPF alone or in combination with a decrease in the glomerular transcapillary hydraulic pressure gradient (DeltaP). PMID- 11856785 TI - Acute humoral rejection in kidney transplantation: II. Morphology, immunopathology, and pathologic classification. AB - The incidence of acute humoral rejection (AHR) in renal allograft biopsies has been difficult to determine because widely accepted diagnostic criteria have not been established. C4d deposition in peritubular capillaries (PTC) of renal allografts has been proposed as a useful marker for AHR. This study was designed to test the relative value of C4d staining, histology, and serology in the diagnosis of AHR. Of 232 consecutive kidney transplants performed at a single institution from July 1995 to July 1999, all patients (n = 67) who developed acute rejection within the first 3 mo and had a renal biopsy with available frozen tissue at acute rejection onset, as well as posttransplant sera within 30 d of the biopsy, were included in this study. Hematoxylin and eosin and periodic acid-Schiff stained sections were scored for glomerular, vascular, and tubulointerstitial pathology. C4d staining of cryostat sections was done by a sensitive three-layer immunofluorescence method. Donor-specific antibodies (DSA) were detected in posttransplant recipient sera using antihuman-globulin-enhanced T cell and B cell cytotoxicity assays and/or flow cytometry. Widespread C4d staining in PTC was present in 30% (20 of 67) of all acute rejection biopsies. The initial histologic diagnoses of the C4d(+) acute rejection cases were as follows: AHR only, 30%; acute cellular rejection (ACR) and AHR, 45%; ACR (CCTT types 1 or 2) alone, 15%; and acute tubular injury (ATI), 10%. The distinguishing morphologic features in C4d(+) versus C4d(-) acute rejection cases included the following: neutrophils in PTC, 65% versus 9%; neutrophilic glomerulitis, 55% versus 4%; neutrophilic tubulitis, 55% versus 9%; severe ATI, 75% versus 9%; and fibrinoid necrosis in glomeruli, 20% versus 0%, or arteries, 25% versus 0%; all P < 0.01. Mononuclear cell tubulitis was more common in the C4d(-) group (70% versus 100%; P < 0.01). No significant difference between C4d(+) and C4d(-) acute rejection was noted for endarteritis, 25% versus 32%; interstitial inflammation (mean % cortex), 27.2 +/- 27% versus 38 +/- 21%; interstitial hemorrhage, 25% versus 15%; or infarcts, 5% versus 2%. DSA were present in 90% (18 of 20) of the C4d(+) cases compared with 2% (1 of 47) in the C4d(-) acute rejection cases (P < 0.001). The pathology of the C4d(+) but DSA(-) cases was not distinguishable from the C4d(+), DSA(+) cases. The C4d(+) DSA(-) cases may be due to non-HLA antibodies or subthreshold levels of DSA. The sensitivity of C4d staining is 95% in the diagnosis of AHR compared with the donor-specific antibody test (90%). Overall, eight grafts were lost to acute rejection in the first year, of which 75% (6 of 8) had AHR. The 1-yr graft failure rate was 27% (4 of 15) for those AHR cases with only capillary neutrophils versus 40% (2 of 5) for those who also had fibrinoid necrosis of arteries. In comparison, the 1-yr graft failure rates were 3% and 7%, respectively, in ACR 1 (Banff/CCTT type 1) and ACR 2 (Banff/CCTT type 2) C4d(-) groups. A substantial fraction (30%) of biopsy-confirmed acute rejection episodes have a component of AHR as judged by C4d staining; most (90%), but not all, have detectable DSA. AHR may be overlooked in the presence of ACR or ATI by histology or negative serology, arguing for routine C4d staining of renal allograft biopsies. Because AHR has a distinct therapy and prognosis, we propose that it should be classified separately from ACR, with further sub-classification into AHR 1 (neutrophilic capillary involvement) and AHR 2 (arterial fibrinoid necrosis). PMID- 11856787 TI - Anemia and iron deficiencies among long-term renal transplant recipients. AB - Iron deficiency anemia after renal transplantation has not been systematically investigated. The prevalence of anemia and the indicators of iron deficiency among 438 renal transplant recipients were examined. Anemia was present in 39.7% of the patients. The prevalence of iron deficiencies, as indicated by a percentage of hypochromic red blood cells (HRBC) of >or=2.5%, was 20.1%. The majority of severely anemic patients exhibited HRBC values in the upper quartile. Positive associations of hemoglobin levels with creatinine clearance, serum transferrin levels, male gender, transferrin saturation (TSAT), polycystic kidney disease, and age were observed. Negative associations with erythropoietin therapy, use of azathioprine, serum ferritin levels, and body mass index were observed. The risk for anemia was closely related to the highest quartile of HRBC percentages (odds ratio, 2.35; 95% confidence interval, 1.48 to 3.75; P = 0.00029), whereas ferritin levels and TSAT conferred no risk for anemia. Therefore, assessment of the HRBC proportion is superior to decreased ferritin and decreased TSAT measurements for the diagnosis of iron deficiencies among renal transplant recipients. PMID- 11856786 TI - Glomerular-specific gene excision in vivo. AB - Podocytes (glomerular visceral epithelial cells) are highly specialized cells that are found in the renal glomerulus and make up a major portion of the filtration barrier between the blood and urinary spaces. Recently, the identification of a number of genes responsible for both autosomal dominant and recessive forms of human nephrotic syndrome has provided insight into a number of molecules responsible for unique features of the podocyte such as the slit diaphragms. Despite these major advances in our understanding of podocyte biology, the function of many genes expressed in the podocyte remains unknown. Targeted gene disruption using homologous recombination in murine embryonic stem cells (ES cells) is a powerful tool to determine the biologic function of genes in vivo. However, resulting embryonic lethal or pleiotropic phenotypes often preclude the analysis of genes in specific renal cell types. To overcome this problem, a glomerular-specific Cre-recombinase transgenic murine line under the control of the Nphs1 (nephrin) promoter (Neph-Cre) was generated. This article reports successful Cre-mediated excision of a 'floxed' transgene specifically in podocytes in vivo. This murine founder line represents a powerful new tool for the manipulation of the expression of genes in podocytes and will provide valuable insight into podocyte biology in the whole animal. PMID- 11856788 TI - New insights into diuretic use in patients with chronic renal disease. AB - Patients with chronic renal insufficiency (CRI) or the nephrotic syndrome frequently manifest diuretic resistance. Factors limiting diuretic responsiveness in patients with CRI may include a reduced basal level of fractional Na(+) reabsorption that places an upper limit on diuretic response, and enhanced NaCl reabsorption in downstream segments, combined with a reduced delivery of diuretic to the kidney. Diuretics are secreted by the recently characterized organic anion transporters (OATs), which are expressed in proximal tubule cells. Secretion may be inhibited by retained organic anions, urate, or acidosis. These limitations necessitate an increased diuretic dosage, up to a defined ceiling level, and consideration of the use of a nonrenally metabolized loop diuretic rather than furosemide. Diuretic responsiveness in patients with the nephrotic syndrome is limited by avid Na(+) reabsorption by the terminal nephron. Experimental studies have shown that a reduced serum albumin concentration can increase the volume of distribution of loop diuretics, reduce their tubular secretion, and enhance the inactivation of furosemide within the kidney by glucuronidization. Binding of loop diuretics can curtail their action in the loop of Henle. Recent clinical investigations have challenged the importance of some of these mechanisms that were identified in animal models. Strategies to improve loop diuretic responsiveness include increasing diuretic dosage, concurrent use of a thiazide diuretic to inhibit downstream NaCl reabsorption and attempts to maximally reduce albumin excretion. Strategies to limit albumin excretion include the use of an angiotensin-converting enzyme inhibitor or an angiotensin receptor blocker and appropriate limitation of protein intake. These measures are more logical, effective, and less expensive than infusion of albumin solutions. PMID- 11856790 TI - Basic fibroblast growth factor and Shiga toxin-O157:H7-associated hemolytic uremic syndrome. PMID- 11856789 TI - Role of the microvascular endothelium in progressive renal disease. AB - The role of the vascular endothelium in progressive renal disease is not well understood. This review presents evidence that progressive renal disease is characterized by a progressive loss of the microvasculature. The loss of the microvasculature correlates directly with the development of glomerular and tubulointerstitial scarring. The mechanism is mediated in part by a reduction in the endothelial proliferative response, and this impairment in capillary repair is mediated by alteration in the local expression of both angiogenic (vascular endothelial growth factor) and antiangiogenic (thrombospondin 1) factors in the kidney. The alteration in balance of angiogenic growth factors is mediated by both macrophage-associated cytokines (interleukin-1beta) and vasoactive mediators. Finally, there is intriguing evidence that stimulation of angiogenesis and/or capillary repair may stabilize renal function and slow progression and that this benefit occurs independently of effects on BP or proteinuria. Therefore, angiogenic agents may represent a novel therapeutic approach for slowing the progression of renal disease. PMID- 11856791 TI - Chemokines and transplant immunobiology. PMID- 11856792 TI - Hypothermia to protect the brain. PMID- 11856793 TI - Mild therapeutic hypothermia to improve the neurologic outcome after cardiac arrest. AB - BACKGROUND: Cardiac arrest with widespread cerebral ischemia frequently leads to severe neurologic impairment. We studied whether mild systemic hypothermia increases the rate of neurologic recovery after resuscitation from cardiac arrest due to ventricular fibrillation. METHODS: In this multicenter trial with blinded assessment of the outcome, patients who had been resuscitated after cardiac arrest due to ventricular fibrillation were randomly assigned to undergo therapeutic hypothermia (target temperature, 32 degrees C to 34 degrees C, measured in the bladder) over a period of 24 hours or to receive standard treatment with normothermia. The primary end point was a favorable neurologic outcome within six months after cardiac arrest; secondary end points were mortality within six months and the rate of complications within seven days. RESULTS: Seventy-five of the 136 patients in the hypothermia group for whom data were available (55 percent) had a favorable neurologic outcome (cerebral performance category, 1 [good recovery] or 2 [moderate disability]), as compared with 54 of 137 (39 percent) in the normothermia group (risk ratio, 1.40; 95 percent confidence interval, 1.08 to 1.81). Mortality at six months was 41 percent in the hypothermia group (56 of 137 patients died), as compared with 55 percent in the normothermia group (76 of 138 patients; risk ratio, 0.74; 95 percent confidence interval, 0.58 to 0.95). The complication rate did not differ significantly between the two groups. CONCLUSIONS: In patients who have been successfully resuscitated after cardiac arrest due to ventricular fibrillation, therapeutic mild hypothermia increased the rate of a favorable neurologic outcome and reduced mortality. PMID- 11856794 TI - Treatment of comatose survivors of out-of-hospital cardiac arrest with induced hypothermia. AB - BACKGROUND: Cardiac arrest outside the hospital is common and has a poor outcome. Studies in laboratory animals suggest that hypothermia induced shortly after the restoration of spontaneous circulation may improve neurologic outcome, but there have been no conclusive studies in humans. In a randomized, controlled trial, we compared the effects of moderate hypothermia and normothermia in patients who remained unconscious after resuscitation from out-of-hospital cardiac arrest. METHODS: The study subjects were 77 patients who were randomly assigned to treatment with hypothermia (with the core body temperature reduced to 33 degrees C within 2 hours after the return of spontaneous circulation and maintained at that temperature for 12 hours) or normothermia. The primary outcome measure was survival to hospital discharge with sufficiently good neurologic function to be discharged to home or to a rehabilitation facility. RESULTS: The demographic characteristics of the patients were similar in the hypothermia and normothermia groups. Twenty-one of the 43 patients treated with hypothermia (49 percent) survived and had a good outcome--that is, they were discharged home or to a rehabilitation facility--as compared with 9 of the 34 treated with normothermia (26 percent, P=0.046). After adjustment for base-line differences in age and time from collapse to the return of spontaneous circulation, the odds ratio for a good outcome with hypothermia as compared with normothermia was 5.25 (95 percent confidence interval, 1.47 to 18.76; P=0.011). Hypothermia was associated with a lower cardiac index, higher systemic vascular resistance, and hyperglycemia. There was no difference in the frequency of adverse events. CONCLUSIONS: Our preliminary observations suggest that treatment with moderate hypothermia appears to improve outcomes in patients with coma after resuscitation from out-of hospital cardiac arrest. PMID- 11856795 TI - A long-term study of prognosis in monoclonal gammopathy of undetermined significance. AB - BACKGROUND: A monoclonal gammopathy of undetermined significance (MGUS) occurs in up to 2 percent of persons 50 years of age or older. Reliable predictors of progression have not been identified, and information on prognosis is limited. METHODS: We identified 1384 patients residing in southeastern Minnesota in whom MGUS was diagnosed at the Mayo Clinic from 1960 through 1994. The primary end point was progression to multiple myeloma or another plasma-cell cancer. RESULTS: During 11,009 person-years of follow-up, MGUS progressed in 115 of the 1384 patients to multiple myeloma, IgM lymphoma, primary amyloidosis, macroglobulinemia, chronic lymphocytic leukemia, or plasmacytoma (relative risk of progression, 25.0, 2.4, 8.4, 46.0, 0.9, and 8.5, respectively). The overall relative risk of progression was 7.3 in these patients as compared with the white population of the Iowa Surveillance, Epidemiology, and End Results program. In 32 additional patients, the monoclonal protein concentration increased to more than 3 g per deciliter or the percentage of plasma cells in the bone marrow increased to more than 10 percent (smoldering multiple myeloma) but without progression to overt myeloma or related disorders. The cumulative probability of progression was 12 percent at 10 years, 25 percent at 20 years, and 30 percent at 25 years. The initial concentration of serum monoclonal protein was a significant predictor of progression at 20 years. CONCLUSIONS: The risk of progression of MGUS to multiple myeloma or related disorders is about 1 percent per year. PMID- 11856796 TI - Leptin-replacement therapy for lipodystrophy. AB - BACKGROUND: The adipocyte hormone leptin is important in regulating energy homeostasis. Since severe lipodystrophy is associated with leptin deficiency, insulin resistance, hypertriglyceridemia, and hepatic steatosis, we assessed whether leptin replacement would ameliorate this condition. METHODS: Nine female patients (age range, 15 to 42 years; eight with diabetes mellitus) who had lipodystrophy and serum leptin levels of less than 4 ng per milliliter (0.32 nmol per milliliter) received recombinant methionyl human leptin (recombinant leptin). Recombinant leptin was administered subcutaneously twice a day for four months at escalating doses to achieve low, intermediate, and high physiologic replacement levels of leptin. RESULTS: During treatment with recombinant leptin, the serum leptin level increased from a mean (+/- SE) of 1.3 +/- 0.3 ng per milliliter to 11.1 +/- 2.5 ng per milliliter (0.1 +/- 0.02 to 0.9 +/- 0.2 nmol per milliliter). The absolute decrease in the glycosylated hemoglobin value was 1.9 percent (95 percent confidence interval, 1.1 to 2.7 percent; P=0.001) in the eight patients with diabetes. Four months of therapy decreased average triglyceride levels by 60 percent (95 percent confidence interval, 43 to 77 percent; P<0.001) and liver volume by an average of 28 percent (95 percent confidence interval, 20 to 36 percent; P=0.002) in all nine patients and led to the discontinuation of or a large reduction in antidiabetes therapy. Self-reported daily caloric intake and the measured resting metabolic rate also decreased significantly with therapy. Overall, recombinant leptin therapy was well tolerated. CONCLUSIONS: Leptin replacement therapy improved glycemic control and decreased triglyceride levels in patients with lipodystrophy and leptin deficiency. Leptin deficiency contributes to the insulin resistance and other metabolic abnormalities associated with severe lipodystrophy. PMID- 11856797 TI - Images in clinical medicine. Coronary spasm. PMID- 11856798 TI - Strategies to improve long-term outcomes after renal transplantation. PMID- 11856799 TI - Obesity. PMID- 11856801 TI - Therapeutic hypothermia after cardiac arrest. PMID- 11856800 TI - Case records of Massachusetts General Hospital. Weekly clinicopathological exercises. Case 6-2002. A 54-year-old woman with left, then right, central retinal-vein occlusion. PMID- 11856802 TI - Genes, microbes, and T cells--new therapeutic targets in Crohn's disease. PMID- 11856803 TI - Blood and disaster--supply and demand. PMID- 11856804 TI - Chemotherapy in the elderly. PMID- 11856805 TI - Tuberculosis and treatment with infliximab. PMID- 11856806 TI - Lack of health insurance and overall health. PMID- 11856807 TI - Research in developing countries. PMID- 11856808 TI - Nongenetic male pseudohermaphroditism. PMID- 11856809 TI - Reactions to the events of September 11. PMID- 11856810 TI - Emergence of macrolide resistance during treatment of pneumococcal pneumonia. PMID- 11856811 TI - Medicaid. PMID- 11856812 TI - Growth hormone-induced alterations in the insulin-signaling system. AB - Growth hormone (GH) counteracts insulin action on lipid and glucose metabolism. However, the sequence of molecular events leading to these changes is poorly understood. Insulin action is initiated by binding of the hormone to its cell surface receptor (IR). This event activates the intrinsic tyrosine kinase activity residing in the beta-subunit of the IR and leads to autophosphorylation of the cytoplasmic portion of the beta-subunit and further activation of its tyrosine kinase towards several intermediate proteins, including the family of IR substrates (IRS) and the Shc proteins. When tyrosine phosphorylated, these cellular substrates connect the IR with several downstream signaling molecules. One of them is the enzyme phosphatidylinositol (PI) 3-kinase. The insulin antagonistic action of GH is not a consequence of a direct interaction with the IR. Instead, long-term exposure to GH is, in general, associated with hyperinsulinemia, which leads to a reduction of IR levels and an impairment of its tyrosine kinase activity. The signals of GH and insulin may converge at post receptor levels. The signaling pathway leading to activation of PI 3-kinase appears to be an important site of convergence between the signals of these two hormones and seems to be mediated principally by IRS-1. Rodent models of chronic GH excess have been useful tools to investigate the mechanism by which GH induces insulin resistance. Decreased IR, IRS-1, and IRS-2 tyrosyl phosphorylation in response to insulin was found in skeletal muscle, whereas a chronic activation of the IRS-PI 3-kinase pathway was found in liver. The induction of the expression of proteins that inhibit IR signaling such as suppressors of cytokine signaling (SOCS)-1 and -6 may also be involved in this alteration. Interestingly, the modulation of insulin signaling and action observed in states of GH excess, deficiency, or resistance seems to be relevant to the changes in longevity associated with those states. PMID- 11856813 TI - Alloantigen system L affects the outcome of rous sarcomas. AB - This study was designed to examine the alloantigen system L effects on Rous sarcomas in three B complex genotypes. The parental stock was 50% Modified Wisconsin Line 3 x White Leghorn Line NIU 4 and 50% inbred Line 6.15-5. Pedigree matings of two B(2)B(5) L(1)L(2) sires to five B(2)B(5) L(1)L(2) dams per sire produced experimental chicks segregating for B and L genotypes. Chicks were inoculated with 20 pock-forming units (pfu) of Rous sarcoma virus (RSV) at 6 weeks of age. Tumors were scored six times over 10 weeks postinoculation after which the tumor scores were used to assign a tumor profile index (TPI) to each chicken. Tumor growth over time and TPI were evaluated by repeated-measures analysis of variance and analysis of variance, respectively. Six trials were conducted with a total of 151 chickens. The major histocompatibility (B) complex affected the responses as the B(2)B(2) and B(2)B(5) genotypes had significantly lower tumor growth over time and TPI than the B(5)B(5) genotype. Separate analyses revealed no significant L system effect in B(2)B(2) or B(2)B(5) backgrounds. However, L genotype significantly affected (P < 0.05) both tumor growth over time and TPI in B(5)B(5) chickens. B(5)B(5) L(1)L(2) birds had TPI significantly lower than B(5)B(5) L(1)L(1) chickens but not B(5)B(5) L(2)L(2). Mortality was lower in the B(5)B(5) L(1)L(2) birds than in B(5)B(5) L(2)L(2) chickens. The L system, or one closely linked, affects the growth and ultimate outcome of Rous sarcomas. The response may depend upon the genetic background as well as MHC type. PMID- 11856814 TI - Plasma glucagon and free fatty acid responses to a glucose load in the obese spontaneous hypertensive rat (SHROB) model of metabolic syndrome X. AB - Metabolic Syndrome X is a cluster of abnormalities including insulin resistance, hyperlipidemia, hypertension, and obesity. We sought to determine if excess plasma glucagon and free fatty acids (FFA) might contribute to the insulin resistance in the obese spontaneous hypertensive rat (SHROB), a unique animal model of leptin resistance and metabolic Syndrome X. SHROB were extremely hyperinsulinemic and mildly glucose intolerant compared with lean SHR. SHROB had elevated fasting plasma glucagon and FFA, and showed paradoxical responses to an oral glucose challenge, with increased glucagon at 30 and 60 min postchallenge (200% plus minus 45% and 91% plus minus 13%, respectively; n = 9). In lean SHR, glucagon was nearly unchanged by glucose loading (<30% increase, P > 0.05; n = 5). Plasma FFA were not affected by a glucose load in SHROB, whereas SHR showed a decrease of 40% plus minus 6% (n = 5--9). The I/G molar ratio changed in opposite directions in the two genotypes, with a decrease in SHROB at 30 and 60 min, in contrast to the appropriate increase at 30 and 60 min postchallenge in the lean SHR (P < 0.01; n = 5--9). Administration of 500 ng/kg exogenous glucagon to SHR raised glucagon 56% plus minus 5% to a level that was similar to fasting SHROB. This level of circulating glucagon was sufficient to elevate glucose and insulin during the 7 hr of observation (n = 9). Based on these results, we suggest that fasting hyperglucagonemia and impaired suppression of glucagon secretion and FFA in response to an oral glucose load may contribute to insulin resistance and glucose intolerance in the SHROB model of metabolic Syndrome X. PMID- 11856815 TI - TGF-beta1 is an autocrine mediator of renal tubular epithelial cell growth and collagen IV production. AB - Recent studies in cultured cells have provided evidence that a variety of pathobiologic stimuli, including high glucose, angiotensin II, and thromboxane A(2), trigger a signaling pathway leading to autocrine induction of TGF-beta1. TGF-beta1 production through this pathway may profoundly affect cell growth, matrix synthesis, and response to injury. This study examines the role of autocrine versus exogenously added TGF-beta1 in cellular proliferation and collagen IV production, critical targets of TGF-beta1 signaling, using renal cells derived from TGF-beta1 knockout (KO) animals or wild-type (WT) controls. Growth of WT and KO cells was assessed by cell counting and [(3)H]thymidine uptake. Basal and TGF-beta1-stimulated collagen production was assessed by Northern and Western blotting; transcriptional activity of the alpha1(IV) collagen gene was assessed by transient transfection analysis. KO cells grew at a faster rate than WT cells carefully matched for plating density and passage number. This increased growth rate was paralleled by increases in [(3)H]thymidine uptake. KO cells expressed lower levels of the cell cycle inhibitors p21 and p27 than WT cells. KO cells failed to express TGF-beta1, as expected. Basal TGF-beta3 mRNA levels were higher in KO cells than in WT cells. WT cells expressed higher basal levels of TGF-beta2 mRNA than KO cells. Basal alpha1(IV) and alpha2(IV) collagen mRNA and protein expression were significantly lower in KO cells than WT cells. Administration of exogenous TGF-beta1 induced collagen IV production in both KO and WT cells. Although basal transcriptional activity of an alpha1(IV) collagen-CAT construct was lower in KO cells than WT cells, administration of exogenous TGF-beta1 was associated with significant increases in transcriptional activity of this construct in both KO and WT cells. These studies provide evidence that autocrine production of TGF-beta1 may play a critical role in regulation of growth and basal collagen IV production by renal tubular epithelial cells. PMID- 11856816 TI - Induction of cardiac cytochrome p450 in cocaine-treated mice. AB - Cytochrome P450 (P450) is a ubiquitous family of enzymes responsible for the metabolism of a wide variety of drugs and their metabolites, including cocaine. To investigate the effects of cocaine on myocardial injuries and cardiac P450 expression, BALB/c mice were injected daily intraperitoneally with cocaine (30 mg/kg) or cocaine plus pretreatment of P450 inhibitors for 14 days. Tumor necrosis factor-alpha (TNF-alpha) content and creatine phosphokinase (CPK) activity in mice hearts and serums were significantly increased after long-term treatment with cocaine. Pretreatment with the P450 inhibitor, cimetidine (Cime, 50 mg/kg) or metyrapone (Mety, 40 mg/kg) abolished or significantly attenuated the effects of cocaine on TNF-alpha and CPK activity. Western blot analysis shows that mouse cardiac tissues express the P450 isoforms CYP1A1, CYP1A2, and CYP2J2. The protein levels normalized with cyclophilin A were 1.20 plus minus 0.07, 0.67 plus minus 0.03, and 1.48 plus minus 0.01 for CYP1A1, CYP1A2, and CYP 2J2, respectively. After cocaine administration, CYP2J2 increased by 43.6% and CYP1A1 increased by 108.5%, but CYP1A2 was not significantly altered. However, the cytochrome P450 inhibitors Cime and Mety suppressed the cocaine-induced increase in CYP1A1 and CYP2J2 expression. Moreover, application of Cime or Mety alone did not alter the level of cardiac TNF-alpha or the expression of P450. Our results demonstrate that long-term exposure to cocaine causes an increase in cardiac CYP1A1 and CYP2J2 concentration. We speculate that induction of P450 isoforms may cause cardiac injury due to cocaine metabolites locally catalyzed by P450 or the increase in P450 expression itself. PMID- 11856817 TI - Transplacental carbohydrate and sugar alcohol concentrations and their uptakes in ovine pregnancy. AB - The concentrations of glucose, fructose, sorbitol, glycerol, and myo-inositol in sheep blood and tissues have been reported previously (1--5). However, the other polyols that are at low concentrations have not been investigated in pregnant sheep due to technical difficulties. By using HPLC and gas chromatography-mass spectrometry, seven polyols (myo-inositol, glycerol, erythritol, arabitol, sorbitol, ribitol, and mannitol) and three hexoses (mannose, glucose, and fructose) were identified and quantified in four blood vessels supplying and draining the placenta (maternal artery, uterine vein, fetal artery, and umbilical vein). Uterine and umbilical blood flows were measured, and uptakes of all the polyols and hexoses in both maternal and fetal circulations were calculated. There was a significant net placental release of sorbitol to both maternal and fetal circulations. Fructose was also taken up significantly by the uterine circulation. Maternal plasma mannose concentrations were higher than fetal concentrations, and there was a net umbilical uptake of mannose, characteristics that are similar to those of glucose. Myo-inositol and erythritol had relatively high concentrations in fetal plasma (697.8 plus minus 53 microM and 463.8 plus minus 27 microM, respectively). The ratios of fetal/maternal plasma arterial concentrations were very high for most polyols. The concentrations of myo inositol, glycerol, and sorbitol were also high in sheep placental tissue (2489 plus minus 125 microM/kg wet tissue, 2119 plus minus 193 microM/kg wet tissue, and 3910 plus minus 369 microM/kg wet tissue), an indication that these polyols could be made within the placenta. PMID- 11856818 TI - Decreased expression of alpha2,8 sialyltransferase and increased expression of beta1,4 N-acetylgalactosaminyltransferase in gastrointestinal cancers. AB - Gangliosides such as GD3, GM2, and GD2 are abundantly expressed on the cell surfaces of various malignant cells, suggesting the potential for anti ganglioside antibody therapy for tumors. Anti-ganglioside GD2 antibody treatment is currently undergoing clinical trials for melanoma and neuroblastoma. We previously reported high in vivo antitumor effects of anti-GM2 ganglioside antibody against lung cancer. To determine whether anti-GM2 antibody may be clinically indicated for gastrointestinal cancers, we evaluated the mRNA expression of alpha2,8 sialyltransferase, a GD3 synthase, and beta1,4 N acetylgalactosaminyltransferase (beta1,4 GalNAc-T), a GM2/GD2 synthase, in gastrointestinal cancers. We performed modified semi-quantitative RT-PCR, which reduces complexity incidental to radiolabeling on samples taken from small surgically removed clinical specimens. Stomach (19/22) and colorectal (21/30) cancers showed decreased expression of alpha2,8 sialyltransferase as compared with respective normal tissues (P < 0.05). In contrast, increased expression of beta1,4 GalNAc-T was detected in both types of tumors. Clinicopathological analysis revealed significantly higher expression level of alpha2,8 sialyltransferase in the poorly differentiated than in the well-differentiated stomach cancer group (P < 0.05). Furthermore, the expression level of alpha2,8 sialyltransferase was significantly decreased in male as compared with female colorectal cancer patients (P < 0.05). These results suggest that expression level of GM2 ganglioside is elevated in gastrointestinal cancer, and that anti GM2 antibody may be applicable to its treatment. PMID- 11856819 TI - Melatonin fails to modulate immune parameters influenced by calorie restriction in aging Fischer 344 rats. AB - The aim of this study was to determine if long-term treatment with melatonin (MEL), a purported anti-aging agent, was as effective as calorie restriction (CR) in modulating immune parameters in aging Fischer 344 male rats. Splenic lymphocytes were isolated from 17-month-old rats that, beginning at 6 weeks of age, were treated with MEL (4 or 16 microg/ml in drinking water) and from 17 month-old rats fed ad libitum (AL) or rats fed a CR diet (55% of AL intake). The number of splenic T cell populations and T cell subsets was measured by flow cytometry, the proliferative response of splenocytes to Concanavalin A (Con A) and lipopolysaccharide (LPS) was measured by [(3)H]thymidine incorporation, and the induction of cytokine production (IL-2 and IFN-gamma) was measured by ELISA assay. In addition, the level of the natural killer (NK) cell activity was assessed by fluorimetric assay. CR rats had a higher number of lymphocytes expressing the naive T cell marker (CD3 OX22) than AL rats (P < 0.05). CR rats also showed greater induction of proliferative response, IL-2 and IFN-gamma levels following Con A simulation, and NK cell activity than AL rats (P < 0.05). MEL-treated rats did not differ from AL rats in any of these parameters or in any other measurement. These results indicate that MEL treatment is unable to modulate immune function in a manner comparable with that of CR. PMID- 11856820 TI - Hypothalamic histamine neurons activate lipolysis in rat adipose tissue. AB - The contribution of hypothalamic histamine neurons to the central regulation of peripheral lipid metabolism was investigated in rats using in vivo microdialysis system. A bolus infusion of L-histamine at doses of 10--10(3) nmol/rat into the third cerebral ventricle (i3vt) dose-dependently increased glycerol concentration in the perfusate from the epididymal adipose tissue. I3vt infusion of 10(2) nmol/rat thioperamide, an autoinhibitory H(3) receptor antagonist that activates histamine neurons to increase synthesis and release of neuronal histamine, convincingly mimicked histamine action in the augmented lipolysis. Intraperitoneal pretreatment with propranolol, a beta-adrenoceptor antagonist, abolished the thioperamide-induced lipolytic action. An electrophysiological study demonstrated that efferent sympathetic nerves innervating the epididymal fat were activated after the i3vt infusion of thioperamide. Hypothalamic histamine neurons thus regulate peripheral lipid metabolism through the accelerating lipolytic action by activation of sympathetic beta-adrenoceptor. PMID- 11856821 TI - Metallothionein protection against alcoholic liver injury through inhibition of oxidative stress. AB - Antioxidants are likely potential pharmaceutical agents for the treatment of alcoholic liver disease. Metallothionein (MT) is a cysteine-rich protein and functions as an antioxidant. This study was designed to determine whether MT confers resistance to acute alcohol-induced hepatotoxicity and to explore the mechanistic link between oxidative stress and alcoholic liver injury. MT overexpressing transgenic and wild-type mice were administrated three gastric doses of alcohol at 5 g/kg. Liver injury, oxidative stress, and ethanol metabolism-associated changes were determined. Acute ethanol administration in the wild-type mice caused prominent microvesicular steatosis, along with necrosis and elevation of serum alanine aminotransferase. Ultrastructural changes of the hepatocytes include glycogen and fat accumulation, organelle abnormality, and focal cytoplasmic degeneration. This acute alcohol hepatotoxicity was significantly inhibited in the MT-transgenic mice. Furthermore, ethanol treatment decreased hepatic-reduced glutathione, but increased oxidized glutathione along with lipid peroxidation, protein oxidation, and superoxide generation in the wild type mice. This hepatic oxidative stress was significantly suppressed in the MT transgenic mice. However, MT did not affect the ethanol metabolism-associated decrease in NAD(+)/NADH ratio or increase in cytochrome P450 2E1. In conclusion, MT is an effective agent in cytoprotection against alcohol-induced liver injury, and hepatic protection by MT is likely through inhibition of alcohol-induced oxidative stress. PMID- 11856822 TI - The structure of the FERM domain of merlin, the neurofibromatosis type 2 gene product. AB - Neurofibromatosis type 2 is an autosomal dominant disorder characterized by central nervous system tumors. The cause of the disease has been traced to mutations in the gene coding for a protein that is alternately called merlin or schwannomin and is a member of the ERM family (ezrin, radixin and moesin). The ERM proteins link the cytoskeleton to the cell membrane either directly through integral membrane proteins or indirectly through membrane-associated proteins. In this paper, the expression, purification, crystallization and crystal structure of the N-terminal domain of merlin are described. The crystals exhibit the symmetry of space group P2(1)2(1)2(1), with two molecules in the asymmetric unit. The recorded diffraction pattern extends to 1.8A resolution. The structure was solved by the molecular-replacement method and the model was refined to a conventional R value of 19.3% (R(free) = 22.7%). The N-terminal domain of merlin closely resembles those described for the corresponding domains in moesin and radixin and exhibits a cloverleaf architecture with three distinct subdomains. The structure allows a better rationalization of the impact of selected disease causing mutations on the integrity of the protein. PMID- 11856823 TI - Differential effects of short affinity tags on the crystallization of Pyrococcus furiosus maltodextrin-binding protein. AB - Pyrococcus furiosus maltodextrin-binding protein readily forms large orthorhombic crystals that diffract to high resolution. This protein was used as a model system to investigate the influence of five short affinity tags (His(6), Arg(5), Strep tag II, FLAG tag and the biotin acceptor peptide) on the formation of protein crystals and their ability to diffract X-rays. The results indicate that the amino-acid sequence of the tag can have a profound effect on both of these parameters. Consequently, the ability to obtain diffracting crystals of a particular protein may depend as much on which affinity tag is selected as it does on whether an affinity tag is used at all. PMID- 11856824 TI - Three-dimensional structure of the type III secretion chaperone SycE from Yersinia pestis. AB - Many bacterial pathogens utilize a type III (contact-dependent) secretion system to inject cytotoxic effector proteins directly into host cells. This ingenious mechanism, designed for both bacterial offense and defense, has been studied most extensively in Yersinia spp. To be exported efficiently, at least three of the effectors (YopE, YopH and YopT) and several other proteins that transit the type III secretion pathway in Yersinia (YopN, YopD and YopB) must first form transient complexes with cognate-specific Yop chaperone (Syc) proteins. The cytotoxic effector YopE, a selective activator of mammalian Rho-family GTPases, associates with SycE. Here, the structure of Y. pestis SycE at 1.95A resolution is reported. SycE possesses a novel fold with an unusual dimerization motif and an intriguing basic cavity located on the dyad axis of the dimer that may participate in its interaction with YopE. PMID- 11856825 TI - The structure of S100A12 in a hexameric form and its proposed role in receptor signalling. AB - S100A12 is a member of the S100 subfamily of EF-hand calcium-binding proteins; it has been shown to be one of the ligands of the 'receptor for advanced glycation end products' (RAGE) that belongs to the immunoglobulin superfamily and is involved in diabetes, Alzheimer's disease, inflammation and tumour invasion. The structure of the dimeric form of native S100A12 from human granulocytes in the presence of calcium in space group R3 has previously been reported. Here, the structure of a second crystal form in space group P2(1) (unit-cell parameters a = 53.9, b = 100.5, c = 112.7A, beta = 94.6 degrees) solved at 2.7A resolution by molecular replacement using the R3 structure as a search model is reported. Like most S100 proteins, S100A12 is a dimer. However, in the P2(1) crystal form dimers of S100A12 are arranged in a spherical hexameric assembly with an external diameter of about 55 A stabilized by calcium ions bound between adjacent dimers. The putative target-binding sites of S100A12 are located at the outer surface of the hexamer, making it possible for the hexamer to bind several targets. It is proposed that the S100A12 hexameric assembly might interact with three extracellular domains of the receptor, bringing them together into large trimeric assemblies. PMID- 11856826 TI - Re-refinement using reprocessed data to improve the quality of the structure: a case study involving garlic lectin. AB - The structure of dimeric garlic lectin was previously determined to an effective resolution of 2.8A using X-ray intensity data processed by the XDS package and refined using X-PLOR [Chandra et al. (1999), J. Mol. Biol. 285, 1157--1168]. Repeated attempts to grow better crystals with a view to improving the definition of the structure did not succeed. The available raw data were then reprocessed using DENZO. The structure was re-refined with both X-PLOR and CNS separately using the reprocessed data, which extended to a resolution of 2.2A. These two sets of refinements and the two sets using the XDS-processed data afforded an opportunity to compare the performance of different data-processing and refinement packages when dealing with data from weakly diffracting crystals. The best results were obtained when CNS was employed for refinement using data processed by DENZO. The quality and the resolution of the map and the definition of the structure improved substantially. In particular, the amino-acid residues at the variable locations in the sequence, and hence the isolectins, could be identified with a high degree of confidence. It could be established that the crystal asymmetric unit contains two identical heterodimers. The new refined structure also provided a better definition of other finer structural details. PMID- 11856827 TI - The structure of AhrC, the arginine repressor/activator protein from Bacillus subtilis. AB - In the Gram-positive bacterium Bacillus subtilis the concentration of the amino acid L-arginine is controlled by the transcriptional regulator AhrC. The hexameric AhrC protein binds in an L-arginine-dependent manner to pseudo palindromic operators within the promoter regions of arginine biosynthetic and catabolic gene clusters. AhrC binding results in the repression of transcription of biosynthetic genes and in the activation of transcription of catabolic genes. The crystal structure of AhrC has been determined at 2.7 A resolution. Each subunit of the protein has two domains. The C-terminal domains are arranged with 32 point-group symmetry and mediate the major intersubunit interactions. The N terminal domains are located around this core, where they lie in weakly associated pairs but do not obey strict symmetry. A structural comparison of AhrC with the arginine repressor from the thermophile B. stearothermophilus reveals close similarity in regions implicated in L-arginine binding and DNA recognition, but also reveals some striking sequence differences, especially within the C terminal oligomerization domain, which may contribute to the different thermostabilities of the proteins. Comparison of the crystal structure of AhrC with a 30 A resolution model obtained by combining X-ray structure-factor amplitudes with phases derived from electron-microscopic analyses of AhrC crystals confirms the essential accuracy of the earlier model and suggests that such an approach may be more widely useful for obtaining low-resolution phase information. PMID- 11856830 TI - Structure of a Cys25-->Ser mutant of human cathepsin S. AB - Cathepsin S (EC 3.4.22.27), a cysteine proteinase of the papain superfamily, plays a critical role in the generation of a major histocompatibility complex (MHC) class II restricted T-cell response by antigen-presenting cells. Therefore, selective inhibition of this enzyme may be useful in modulating class II restricted T-cell responses in immune-related disorders such as rheumatoid arthritis, multiple sclerosis and extrinsic asthma. The three-dimensional structure at 2.2 A resolution of the active-site Cys25-->Ser mutant presented here in an unliganded state provides further insight useful for the design of selective enzyme inhibitors. PMID- 11856828 TI - Description of ordered solvent molecules in a platinated decanucleotide duplex refined at 1.6A resolution against experimental MAD phases. AB - Accurate experimental phases derived from a MAD experiment may be useful to enable the identification of solvent molecules during the course of an atomic parameter refinement. The structure of a double-stranded DNA decanucleotide bearing a cisplatin interstrand cross-link at 1.6A resolution, whose phases were first determined experimentally using the L(III) edge of the Pt atom, was refined by various methods. The previously published structure resulted from a least squares refinement using the structure-factor magnitudes and stereochemical restraints (program SHELX). In this paper, these previous results are compared with a model obtained by the likelihood-maximization method (program REFMAC) which allows the combination of the observed magnitudes with experimental MAD phases. This solution corresponded to a lower R(free) (18.8 compared with 20.3%), a lower R factor and accounted for 135 water molecules and one spermine molecule collected by the program wARP during refinement. The previously published SHELX solution exhibited no spermine molecule and accounted for 92 water molecules, only 74 of which are also present in the model obtained with the MAD phases. In order to verify that these improvements were actually related to the use of the MAD phases, the same type of procedure without the MAD phases was applied starting from the initial model. The resulting solution had a higher R(free) (20.3%), which could be related to the loss of 22 water molecules and the addition of 20 new ones. MAD phases therefore seem especially helpful in preventing the model bias which may affect the solvent molecules. All models have in common a hydration cage of nine water molecules which surround the platinum residue. In addition to the spermine molecule, the model obtained with the MAD phases allows description of the water-molecule organization, with reproducible motifs related to the base pairs and to the phosphodiester backbone. PMID- 11856829 TI - Atomic resolution structures of ribonuclease A at six pH values. AB - The diffraction pattern of protein crystals extending to atomic resolution guarantees a very accurate picture of the molecular structure and enables the study of subtle phenomena related to protein functionality. Six structures of bovine pancreatic ribonuclease at the pH* values 5.2, 5.9, 6.3, 7.1, 8.0 and 8.8 and at resolution limits in the range 1.05-1.15A have been refined. An overall description of the six structures and several aspects, mainly regarding pH triggered conformational changes, are described here. Since subtle variations were expected, a thorough validation assessment of the six refined models was first carried out. Some stereochemical parameters, such as the N[bond]C(alpha)[bond]C angle and the pyramidalization at the carbonyl C atoms, indicate that the standard target values and their weights typically used in refinement may need revision. A detailed comparison of the six structures has provided experimental evidence on the role of Lys41 in catalysis. Furthermore, insights are given into the structural effects related to the pH-dependent binding of a sulfate anion, which mimics the phosphate group of RNA, in the active site. Finally, the results support a number of thermodynamic and kinetic experimental data concerning the role of the disulfide bridge between Cys65 and Cys72 in the folding of RNase A. PMID- 11856831 TI - Resolution improvement from 'in situ annealing' of copper nitrite reductase crystals. AB - Significant improvement of the resolution of copper nitrite reductase crystals was achieved by using the in situ annealing technique. The effective resolution limits increased by 1.5 A from 2.5 to 1.0 A, the mosaicity value decreased from 1.5 to 0.3 degree and the spot shape changed from elliptical to circular. PMID- 11856832 TI - Flash-cooling and annealing of protein crystals. AB - Flash-cooling and annealing of macromolecular crystals have been investigated using in situ X-ray imaging, diffraction-peak lineshape measurements and conventional crystallographic diffraction. The dominant mechanisms by which flash cooling creates disorder are suggested and a fixed-temperature annealing protocol for reducing this disorder is demonstrated that should be more reliable and flexible than existing protocols. Flash-cooling tetragonal lysozyme crystals degrades diffraction resolution and broadens the distributions of lattice orientations (mosaicity) and lattice spacings. The diffraction resolution strongly correlates with the width of the lattice-spacing distribution. Annealing at fixed temperatures of 253 and 233 K consistently reduces the lattice-spacing spread and improves the resolution for annealing times up to approximately 30s. X ray images show that this improvement arises from the formation of well ordered domains with characteristic sizes >10 microm and narrower mosaicities than the crystal as a whole. Flash-cooled triclinic crystals of lysozyme, which have a smaller water content than the tetragonal form, diffract to higher resolution with smaller mosaicities and exhibit pronounced ordered domain structure even before annealing. It is suggested that differential thermal expansion of the protein lattice and solvent may be the primary cause of flash-cooling-induced disorder. Mechanisms by which annealing at T << 273 K reduce this disorder are discussed. PMID- 11856834 TI - A novel pH-dependent dimerization motif in beta-lactoglobulin from pig (Sus scrofa). AB - beta-Lactoglobulin (BLG) is a lipocalin and is the major protein in the whey of the milk of cows and other ruminants, but not in all mammalian species. The biological function of BLG is not clear, but a potential role in carrying fatty acids through the digestive tract has been proposed. The capability of BLG to aggregate and form gels is often used to thicken foodstuffs. The structure of the porcine form is sufficiently different from other known BLG structures that SIRAS phases had to be measured in order to solve the crystal structure to 2.4 A resolution. The r.m.s. deviation of C(alpha) atoms is 2.8 A between porcine and bovine BLG. Nevertheless, the typical lipocalin fold is conserved. Compared with bovine BLG, the tilted alpha-helix alters the arrangement of surface residues of the porcine form, completely changing the dimerization behaviour. Through a unique pH-dependent domain-swapping mechanism involving the first ten residues, a novel dimer interface is formed at the N-terminus of porcine BLG. The existence of this novel dimer at low pH is supported by gel-filtration experiments. These results provide a rationale for the difference in physicochemical behaviour between bovine and porcine BLG and point the way towards engineering such dimerization motifs into other members of the lipocalin family. PMID- 11856833 TI - X-ray scattering studies of Aspergillus flavus urate oxidase: towards a better understanding of PEG effects on the crystallization of large proteins. AB - The determination of the three-dimensional structures of biological macromolecules by X-ray diffraction generally requires large good-quality crystals, which are often difficult to obtain as crystal nucleation and growth depend upon a great number of physicochemical parameters. In the future, the emergence of structural genomic projects will require new and rapid methods to determine crystallization conditions. Until now, the prediction of crystallization conditions has essentially been based on the knowledge of interparticular interactions in solutions inferred from studies on small soluble proteins in the presence of salts. The present study, by small-angle X-ray scattering, of urate oxidase from Aspergillus flavus, a homotetrameric enzyme of 128kDa, allowed the extension of the results to the crystallization of large proteins in the presence of polyethylene glycol (PEG). The protein crystallization, the nucleation rate and the different morphological crystal shapes obtained were correlated with the second virial coefficient (A(2)), which was found to be in a restricted range at the low end of the 'crystallization slot' proposed by George & Wilson [(1994). Acta Cryst. D50, 361--365]. PMID- 11856835 TI - Pattern-recognition methods to identify secondary structure within X-ray crystallographic electron-density maps. AB - The interpretation of macromolecular crystallographic electron-density maps is a difficult and traditionally a manual step in the determination of a protein structure. The visualization of information within an electron-density map can be extremely arduous owing to the amount and complexity of information present. The ability to see the overall fold and structure of the molecule is usually lost among all the detail, particularly with larger structures. This paper describes a novel method of analysis of electron density in real space that can determine the secondary structure of a protein within minutes without any user intervention. The method is able to work with poor data as well as good data at resolutions down to 3.5A and is integral to the functionality of QUANTA. This article describes the methodology of the pattern recognition and its use with a number of sets of experimental data. PMID- 11856836 TI - Jolly SAD. AB - Examples of phasing macromolecular crystal structures based on single-wavelength anomalous dispersion (SAD) show that this approach is more powerful and may have more general application in structural biology than was anticipated. Better data collection facilities and cryogenic techniques, coupled with powerful programs for data processing, phasing, density modification and automatic model building, means that the SAD approach may gain wide popularity owing to its simplicity, less stringent wavelength requirements and faster data collection and phasing than the multi-wavelength (MAD) approach. It can be performed at any wavelength where anomalous scattering can be observed, in many cases using laboratory X-ray sources. PMID- 11856837 TI - Crystallization and preliminary X-ray diffraction analysis of L-aminoacylase from the hyperthermophilic archaeon Thermococcus litoralis. AB - The enzyme L-aminoacylase catalyses the hydrolysis of N-acyl-L-amino acids from peptides or proteins. The recombinant enzyme from the hyperthermophilic archaeon Thermococcus litoralis has been purified to homogeneity. This zinc-containing enzyme has been crystallized from ammonium sulfate using the sitting-drop vapour diffusion method. The crystals diffract to 2.8 A resolution and belong to the rhombohedral space group R32, with unit-cell parameters a = b = 102.4, c = 178.5 A, gamma = 120 degrees in a hexagonal lattice setting. The asymmetric unit contains one enzyme monomer, containing a single zinc ion. Two synchrotron data sets have been collected at a remote wavelength and at the maximum f'wavelength for zinc. This has allowed the position of the metal to be identified in anomalous Patterson maps. PMID- 11856838 TI - Crystallization of the Oct-1/SNAP190 peptide/DNA complex. AB - Crystals of the Oct-1 POU/SNAP190 peptide/DNA tertiary complex have been obtained by hanging-drop vapor diffusion at 293K in 20% 2-propanol, 20% PEG 4000 and 0.1M sodium citrate pH 5.6. The Oct-1 POU protein has two domains, one a homeodomain and the other a POU domain, which are connected by a flexible linker. The DNA used in the complex is slightly different in the octamer region compared with the two previously crystallized Oct-1 POU/DNA complexes. The DNA is 14 base pairs, with an octamer sequence of 5'-ATGTAGAT-3' and an overhang of one base on both strands. The SNAP190 peptide is 27 amino acids long (residues 884-910). The crystals diffract to 2.3 A (94.1% completeness) at the synchrotron under cryogenic (123K) conditions. The crystals are triclinic, space group P1, with unit-cell parameters a = 36.4, b = 54.9, c = 77.6A, alpha = 94.9, beta = 99.6, gamma = 109.2 degrees. This structure will provide insight into how Oct-1 interacts with SNAP190, a critical component of the small nuclear RNA-activating protein complex (SNAPc). Transcription of human snRNA genes is activated by these direct protein-protein interactions. PMID- 11856839 TI - Crystallization and preliminary X-ray diffraction studies of human angiostatin. AB - Angiostatin is an inhibitor of angiogenesis, the process by which new blood vessels are formed from pre-existing ones. The fact that tumor growth and metastasis dissemination are angiogenesis-dependent processes has awakened interest in angiogenesis inhibitors as anticancer treatment drugs. Angiostatin, a potent angiogenesis inhibitor, is currently in phase I clinical trials. Human angiostatin containing the first three kringle domains of plasminogen (K1-3) has been crystallized and high-resolution data were collected. The crystals belong to the P4(1)2(1)2 space group, with unit-cell parameters a = b = 56.94, c = 192.97 A. A data set to a resolution of 1.75 A with an overall R(merge) and I/sigma(I) of 7% and 19.5, respectively, was obtained. PMID- 11856841 TI - Cloning, purification, crystallization and preliminary X-ray studies of RFC boxes II-VIII of replication factor C from Methanococcus jannaschii. AB - Replication factor C (RFC) is the accessory protein required to load the proliferating cell nuclear antigen (PCNA) onto DNA in replication process. RFC is composed of several subunits and each subunit contains the highly conserved sequences RFC boxes II-VIII. RFC boxes II-VIII of the large subunit of replication factor C from Methanococcus jannaschii has been overexpressed in Escherichia coli, purified and crystallized at 295 K using ammonium sulfate as precipitant. Crystals belong to the space group R32, with unit-cell parameters a = b = 238.23 (5), c = 73.17 (12) A. Native data were collected at 100 K to a resolution of 3.2 A using a synchrotron-radiation source. PMID- 11856840 TI - Crystallization, preliminary X-ray analysis and biophysical characterization of HPr kinase/phosphatase of Mycoplasma pneumoniae. AB - The Mycoplasma pneumoniae HPr kinase/phosphatase (HPrK/P) is a member of a large family of enzymes which are central to carbon regulation in Gram-positive bacteria. The full-length M. pneumonia HPrK/P was crystallized from solutions of polyethylene glycol 8000 and KCl or NaCl which also contained the non hydrolysable ATP analog adenosine 5'-[beta, gamma-methylene]triphosphate (AMPPCP). The crystals belong to the orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a = 117.1, b = 127.7, c = 170.7 A. A complete X-ray intensity data set has been collected and processed to 2.50 A resolution. The slow self-rotation function revealed the presence of a sixfold axis. Dynamic light-scattering (DLS) experiments indicated a molecular weight of 197 kDa for HPrK/P in the absence of AMPPCP and of 217 kDa in the presence of the ATP analog. Thus, the biophysical and crystallographic data suggest that HPrK/P is a functional hexamer that undergoes an ATP-binding-induced conformational change. PMID- 11856842 TI - Crystallization and preliminary X-ray diffraction analysis of cytochrome c peroxidase from the purple phototrophic bacterium Rhodobacter capsulatus. AB - Bacterial cytochrome c peroxidase (BCCP) from Rhodobacter capsulatus was heterologously expressed in Escherichia coli. It was purified to homogeneity and crystallized using the hanging-drop vapour-diffusion method. Diffraction-quality crystals of the enzyme were obtained under two conditions. The first crystal belonged to space group P2(1), with unit-cell parameters a = 99.2, b = 224.7, c = 167.9 A, beta = 105 degrees, and diffracted to 3.5A resolution. The crystallographic asymmetric unit of these crystals contained ten peroxidase molecules. R. capsulatus BCCP also crystallized in space group P2(1)2(1)2(1), with unit-cell parameters a = 67.2, b = 134.4, c = 167.9 A. These crystals diffracted to 2.7 A resolution and contained four peroxidase molecules per crystallographic asymmetric unit. PMID- 11856843 TI - Crystallization and preliminary X-ray characterization of the acylphosphatase like domain from the Escherichia coli hydrogenase maturation factor HypF. AB - Maturation of prokaryotic hydrogenase involves several protein factors, among which is the accessory protein HypF, which hosts the consensus sequence of acylphosphatases and a sequence motif common to proteins catalyzing O carbamoylations. The specific functions of HypF are largely unknown, although it has been observed that CN(-) and CO ligands at the hydrogenase Ni,Fe active centre originate from carbamoylphosphate. The HypF N-terminal domain (91 residues, acylphosphatase-like domain) has been crystallized in two different crystal forms belonging to the orthorhombic P2(1)2(1)2(1) space group (unit-cell parameters a = 35.5, b = 59.8, c = 87.6 A) and to the rhombohedral space group R32 (unit-cell parameters a = b = 58.1, c = 155.6 A in the hexagonal setting). PMID- 11856844 TI - Crystallization of a complex of Caenorhabditis elegans diadenosine tetraphosphate hydrolase and a non-hydrolysable substrate analogue, AppCH2ppA. AB - The molecule diadenosine tetraphosphate (Ap(4)A) has been suggested to be a component of the cellular response to metabolic stress and/or, via the intracellular Ap(3)A/Ap(4)A ratio, to be involved in differentiation and apoptosis. Thus, the enzyme Ap(4)A hydrolase has a key metabolic role through regulation of the intracellular Ap(4)A levels. Crystals of this enzyme from the nematode Caenorhabditis elegans have been obtained in the presence of a non hydrolysable substrate analogue, AppCH(2)ppA. The crystals belong to space group P2(1), unit-cell parameters a = 57.6, b = 36.8, c = 68.9 A, beta = 114.2 degrees, and diffract to approximately 2.0 A. Determination of the structure of this complex will provide insights into the substrate specificity and catalytic activity of this class of enzymes. PMID- 11856845 TI - Crystallization and synchrotron X-ray diffraction studies of human interleukin 22. AB - Human interleukin-22, a novel member of the cytokine family, has been crystallized in hanging drops using the vapour-diffusion technique. Preliminary X ray diffraction experiments using synchrotron radiation reveal that the protein crystallizes in space group P2(1)2(1)2(1), with unit-cell parameters a = 55.44, b = 61.62, c = 73.43 A, and diffracts beyond 2.00 A resolution. PMID- 11856846 TI - Crystallization and preliminary X-ray crystallographic analysis of beta xylosidase from Thermoanaerobacterium saccharolyticum, a thermophilic anaerobe. AB - beta-Xylosidases are involved in the breakdown of xylans into xylose and belong to either family 39 or 43 of the glycosyl hydrolases. At present, no structural information is available for any member of these families. beta-Xylosidase from the thermophilic anaerobe Thermoanaerobacterium saccharolyticum, a member of glycosyl hydrolase family 39, has been crystallized at 296 K using the hanging drop vapour-diffusion method. The crystal diffracts to 2.4 A resolution with synchrotron X-rays and belongs to space group P4(1)2(1)2 (or P4(3)2(1)2), with unit-cell parameters a = b = 92.75, c = 241.37 A. The asymmetric unit contains two monomers of the recombinant enzyme, giving a corresponding V(M) of 2.21 A(3)Da(-1) and a solvent content of 44.3%. PMID- 11856847 TI - Purification, crystallization and preliminary characterization of an Eph B2/ephrin-B2 complex. AB - Eph receptors and their ephrin ligands are involved in various aspects of cell- cell communication during development, including those of the axon pathfinding processes in the nervous system and cell--cell interactions of the vascular endothelial cells. The recognition and binding properties of the ligand-binding domain of EphB2 receptor and the extracellular domain of ephrin-B2 have been studied and two different cocrystals of their complex have been generated. One crystal form has space group C2, diffracts to 3.5 A and has unit-cell parameters a = 128, b = 88, c = 79 A, beta = 112 degrees. The other crystal form grows in space group P1, has unit-cell parameters a = 78, b = 78, c = 78 A, alpha = 69, beta = 75, gamma = 69 degrees and diffracts to 2.7A. Structure-determination experiments using the latter form are in progress. The structure of the complex will elucidate the chemical nature of the interactions between Eph receptors and ephrins, which would create the possibility of using them as targets for structure-based anticancer-drug development. PMID- 11856848 TI - Expression, purification, refolding and crystallization of the carbohydrate recognition domain of p58/ERGIC-53, an animal C-type lectin involved in export of glycoproteins from the endoplasmic reticulum. AB - p58/ERGIC-53 is a mammalian calcium-dependent lectin that serves as a glycoprotein-sorting receptor between the endoplasmic reticulum (ER) and the Golgi complex. It is a type I transmembrane protein with two lumenal domains, one of which is a carbohydrate-recognition domain (CRD) and homologous to leguminous lectins. The CRD of p58, the rat homologue of human ERGIC-53, was overexpressed in insect cells and Escherichia coli, purified and crystallized using Li(2)SO(4) as a precipitant. The crystals belong to space group I222, with unit-cell parameters a = 49.6, b = 86.1, c = 128.1 A, and contain one molecule per asymmetric unit, corresponding to a packing density of 2.4 A(3)Da(-1). Knowledge of the structure of p58/ERGIC-53 will provide a starting model for understanding receptor-mediated glycoprotein sorting between the ER and the Golgi. PMID- 11856849 TI - Crystallization and preliminary X-ray analysis of the ATP-binding domain of the ABC transporter haemolysin B from Escherichia coli. AB - Haemolysin B (HlyB) is a transmembrane protein which belongs to the superfamily of ABC transporters. In vivo, it mediates the non-classical translocation of the 107 kDa toxin HlyA across both membranes of Escherichia coli together with haemolysin D and the outer membrane protein TolC. The cytosolic ATP-binding domain of HlyB has been overexpressed and purified as an N-terminal His-tag fusion protein. Here, the crystallization of the ATPase domain of HlyB in the presence of ATP is described. A native data set has been obtained at a resolution of 2.8 A. Crystals belong to the primitive tetragonal space group P4(x)2(1)2, where x is very likely to be 1 or 3, with unit-cell parameters a = b = 104.6, c = 125.8 A, alpha = beta = gamma = 90 degrees. PMID- 11856850 TI - Crystallization and X-ray analysis of native and selenomethionyl beta-mannanase Man5A from blue mussel, Mytilus edulis, expressed in Pichia pastoris. AB - The glycohydrolase family 5 beta-mannanase Man5A from Mytilus edulis has been expressed in Pichia pastoris and purified in a form suitable for X-ray crystallographic analysis. Crystals were grown by the hanging-drop technique at 293 K using polyethylene glycol 5000 monomethylether as precipitant and dioxane as additive. The crystals belong to the orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a = 61.8, b = 64.8, c = 90.7 A. Diffraction to 1.4 A resolution has been obtained at 100 K. Expression was also performed in the presence of selenomethionine. The incorporation of SeMet was estimated at 40% by amino-acid analysis and its presence in crystals was confirmed from the X-ray absorption scanning spectrum. PMID- 11856851 TI - Thermophilic alcohol dehydrogenase from the mesophile Entamoeba histolytica: crystallization and preliminary X-ray characterization. AB - The tetrameric NADP(+)-dependent secondary alcohol dehydrogenase from Entamoeba histolytica has been crystallized in its apo form. The crystals belong to space group C222(1), with unit-cell parameters a = 76.89, b = 234.24, c = 96.24 A, and diffract to 1.9 A at liquid-nitrogen temperature. Analysis of the Patterson self rotation function shows that the crystals contain one dimer per asymmetric unit. PMID- 11856852 TI - Crystallization of diaminopimelate decarboxylase from Escherichia coli, a stereospecific D-amino-acid decarboxylase. AB - The final step in lysine biosynthesis in bacteria, the conversion of meso diaminopimelate to L-lysine, is catalyzed by the only known D-amino-acid decarboxylase, diaminopimelate decarboxylase (DDC). The Escherichia coli DDC has been cloned, overexpressed in E. coli with a carboxy-terminal polyhistidine purification tag and crystallized from lithium sulfate. The protein is intensely yellow, owing to the pyridoxal-5'-phosphate cofactor, and is enzymatically active. Large well ordered crystals, belonging to space group P6(1)22 with unit cell parameters a = b = 98.6, c = 177 A, make high-resolution X-ray diffraction studies possible to characterize the residues important in stereospecific decarboxylation and reprotonation during catalytic turnover. PMID- 11856853 TI - Crystallization and preliminary X-ray diffraction studies of a D-lysine-based chiral PNA-DNA duplex. AB - A 10-mer duplex formed between a PNA containing a 'chiral box' of three adjacent D-Lys-based monomers and its complementary DNA strand has been crystallized for the first time. Crystals have been obtained using PEG 8000 as precipitant and cacodylate at pH 6.3 as buffer. The crystals belong to the space group P3(1) or to its enantiomorph P3(2), with unit-cell parameters a = b = 35.00, c = 35.91 A. A complete data set has been collected at the synchrotron source Elettra in Trieste to 1.85 A resolution, using a single frozen crystal. PMID- 11856854 TI - Expression, crystallization and preliminary X-ray diffraction studies on the complete choline-binding domain of the major pneumococcal autolysin. AB - The major pneumococcal autolysin (LytA), a virulence factor of this bacterium, is composed of an amino-terminal catalytic domain plus a carboxyl-terminal choline binding domain (ChBD). This C-terminal domain, responsible for anchorage to the cell wall, is a tandem of six imperfect 20-residue repeats whose precise ends have been difficult to establish by sequence methods. The reported crystal structure of a shortened C-terminal fragment of the protein suggested that it might contain an additional repeat and thus an additional choline-binding site (ChBS). The complete recombinant choline-binding domain of LytA has now been overexpressed in soluble form using a secreting Escherichia coli strain which facilitates purification with a higher yield. It has been crystallized at room temperature using MPD as the main precipitant. The crystals belong to space group P2(1) and diffract to beyond 3.2 A resolution on a synchrotron-radiation source. The molecular-replacement solution indicates that a new ChBS which fits the topology of the solenoid structure is formed in the N-terminal region. PMID- 11856855 TI - Crystallization and preliminary X-ray diffraction analysis of chloroplastic ascorbate peroxidase of tobacco plants. AB - The stromal ascorbate peroxidase of tobacco plants was crystallized by the hanging-drop vapour-diffusion method using polyethylene glycol 4000 as a precipitant. The crystals belong to the orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a = 37.2, b = 76.8, c = 98.8 A. The calculated V(M) value based on a monomer in the asymmetric unit is 2.2 A(3)Da(-1). A data set was successfully collected to 1.6 A resolution from a frozen crystal using synchrotron radiation of wavelength 1.0 A at KEK-PF, Japan. PMID- 11856856 TI - Crystallization and preliminary X-ray crystallographic analysis of malonamidase E2, an amidase signature family member. AB - Malonamidase E2 from Bradyrhizobium japonicum catalyzes the hydrolysis of malonamate. The enzyme belongs to an amidase signature family which has a highly conserved serine- and glycine-rich sequence over a stretch of approximately 45 amino acids. More than 100 known or predicted members belonging to this family, whose biological functions vary widely, can be identified in sequence databases. Although urgently needed, no three-dimensional structure of any protein of this family is yet available. The crystallization of malonamidase E2 was undertaken as a first step toward the goal of providing information on the canonical structure of the amidase signature family. The enzyme was crystallized using the hanging drop vapour-diffusion method at 277 K under two different conditions. One crystal form, which is easier to work with than the other form, belongs to the orthorhombic space group P2(1)2(1)2, with unit-cell parameters a = 104.29, b = 95.58, c = 74.90 A. The unit cell is likely to contain two molecules of MAE2, with a crystal volume per protein mass (V(M)) of 2.045 A(3)Da(-1) and solvent content of about 39.9% by volume. A native data set to 1.8 A resolution was obtained from a flash-cooled crystal using synchrotron radiation. PMID- 11856857 TI - Crystallization and preliminary X-ray crystallographic analysis of SEDL. AB - SEDL (known also as sedlin) is a 140 amino-acid protein with a putative role in endoplasmic reticulum-to-Golgi transport. Several missense mutations and deletion mutations in the SEDL gene, which result in protein truncation by frame shift, are responsible for spondyloepiphyseal dysplasia tarda, a progressive skeletal disorder. The protein is identical to MIP-2A, which was shown to interact physically with c-myc promotor-binding protein 1 (MBP-1) and relieve the regulatory role of MBP-1 as a general transcription repressor. In order to gain insights into the function of SEDL by structural analysis, the protein was overexpressed and crystallized as a first step. SEDL was overexpressed in Escherichia coli and crystallized using the hanging-drop vapour-diffusion method at 298 K. The crystals belong to the orthorhombic space group C222(1), with unit cell parameters a = 46.69, b = 101.30, c = 66.15 A. The unit cell is likely to contain one molecule of SEDL, with a crystal volume per protein mass (V(M)) of 2.36 A(3)Da(-1) and a solvent content of about 47.9% by volume. A native data set to 2.8 A resolution was obtained from a flash-cooled crystal using synchrotron radiation. PMID- 11856858 TI - Structural characterization of a new crystal form of the four-way Holliday junction formed by the DNA sequence d(CCGGTACCGG)2: sequence versus lattice? AB - DNA-strand exchange is a vital step in the recombination process, of which a key intermediate is the four-way DNA Holliday junction formed transiently in most living organisms. Here, the single-crystal structure at a resolution of 2.35 A of such a DNA junction formed by d(CCGGTACCGG)(2), which has crystallized in a more highly symmetrical packing mode to that previously observed for the same sequence, is presented. In this case, the structure is isomorphous to the mismatch sequence d(CCGGGACCGG)(2), which reveals the roles of both lattice and DNA sequence in determining the junction geometry. The helices cross at the larger angle of 43.0 degrees (the previously observed angle for this sequence was 41.4 degrees) as a right-handed X. No metal cations were observed; the crystals were grown in the presence of only group I counter-cations. PMID- 11856859 TI - Assessments of severity and management of acute pancreatitis based on the Santorini Consensus Conference report. PMID- 11856860 TI - Subcutaneously inoculated cells and implanted pancreatic cancer tissue show different patterns of metastases in Syrian golden hamsters. AB - CONTEXT: We studied behavior of the subcutaneously implanted pancreatic tumors and the process of metastasis using syngeneic Syrian golden hamsters. DESIGN: HaP T1, a cell line derived from nitrosamine-induced pancreatic cancer in Syrian golden hamsters was used for this experiment. Thirty-five animals were divided into two groups: subcutaneous cell inoculation and subcutaneous tissue implantation. The tumor tissue was obtained from subcutaneously implanted cancer cells. One month after implantation, the tumors were resected and studied histopathologically. The animals were followed-up weekly by palpation of the peripheral lymph nodes in order to identify local recurrence. After death, necropsy was performed. Liver, lungs and pancreas specimens were taken for histopathogical study and detection of K-ras point mutation using the PCR/RFLP method. RESULTS: The mean survival time in the subcutaneous cell inoculation group was 151+/-17.5 days, and in the subcutaneous tissue implantation group was 137 +/-12.9 days. During the follow-up, 13 subcutaneously cell inoculated hamsters (86.7%) had right axillary lymph node metastasis while subcutaneously tissue implanted hamsters did not show any palpable lymph nodes. After necropsy, 10 of the 20 subcutaneously tissue implanted animals (50%) showed metastases in the lungs at the histopathological level. However, 16 of the 20 subcutaneously tissue implanted animals (80%) showed K-ras point mutation in the lung specimens. The lungs of the animals of the subcutaneous cell inoculation group did not show any metastases. No metastases were found in the liver or the pancreas in either group. CONCLUSION: This study suggests that homologous subcutaneous cell inoculation and subcutaneous tissue implantation models showed completely different patterns of metastasis. These models may aid further research to clarify the mechanisms of metastasis in pancreatic cancer. PMID- 11856861 TI - An immunocytochemical profile of the endocrine pancreas using an occlusive duct ligation model. AB - CONTEXT: Ligation of the pancreatic duct, distally to its confluence into the bile duct has been shown to induce endocrine tissue regeneration. The surplus endocrine tissue formed is presumed to be able to replace pathologically and/or experimentally compromised tissue. OBJECTIVE: This is a quantitative study on the histology of duct ligated pancreas employing immunocytochemistry and computerised morphometry. INTERVENTIONS: Pancreatic duct ligation was performed on 25 groups of six normal Sprague-Dawley rats. Experimental animals were sacrificed at 12 hour intervals from day one to ten post-duct ligation and every 24 hours thereafter to day 14, the pancreas removed, fixed and processed. Six consecutive 3-6 micron serial sections were cut on a rotary hand microtome, floated onto 3 aminopropyl-trimethoxysilan coated slides and alternatively immunocytochemically stained for insulin, glucagon, pancreatic polypeptide and somatostatin. RESULTS: Pancreas transformation between days 1/2 and 3 1/2 was characterised by acinar deletion and the appearance of immunoreactive cells for the primary endocrine hormones. Transdifferentiation of existing endocrine tissue saw islet insulin core cells replaced by pancreatic polypeptide- and somatostatin positive cells, glucagon deletion and random appearance of all endocrine cell types within the inter-islet interstitium by day 3 1/2. Days 4 to 14 were characterised by cellular migration and islet reconstruction. CONCLUSIONS: To date our laboratory has investigated transplantation of foetal tissue beneath the renal capsule in syngeneic, isogeneic and allogeneic normal and diabetic rats. As pancreatic duct ligation induces the development of surplus endocrine tissue our next step would be to investigate the use of ligated pancreas as a replacement for foetal tissue. PMID- 11856862 TI - Severe hypophosphatemia in a patient with acute pancreatitis. AB - CONTEXT: We describe a patient with alcohol-induced pancreatitis who developed severe life-threatening hypophosphatemia of multifactorial origin during hospitalization. CASE REPORT: Decreased phosphate levels along with urine phosphate wasting were already noticed on the patient's admission due to underlying chronic alcoholism. However, a further deterioration of hypophosphatemia appeared on the second day of hospitalization presumably resulting from an increased transfer of phosphate from extracellular to intracellular fluid. CONCLUSIONS: Phosphate deficiency is often overlooked in patients with acute pancreatitis. Our case emphasizes that serum phosphate levels should be checked along with serum calcium levels in patients with acute pancreatitis, especially in alcoholic patients. PMID- 11856863 TI - Microvascular complications in cystic fibrosis-related diabetes mellitus: a case report. AB - CONTEXT, The prevalence of cystic fibrosis-related diabetes mellitus is increasing and is associated with increased survival from cystic fibrosis. CASE REPORT, This study describes a case of the premature onset of disabling and widespread microvascular complications resulting from cystic fibrosis-related diabetes mellitus. Previously asymptomatic retinopathy was diagnosed on recognition of diabetic nephropathy. CONCLUSIONS, The treatment of pulmonary exacerbations has become more complex due to the nephrotoxic potential of intravenous aminoglycoside drugs which are frequently used to control chronic Pseudomonas infection in cystic fibrosis. PMID- 11856864 TI - New Journal name, new section, new brief gene mapping report policy. Starting in 2002 the name of this Journal will be: Cytogenetic and Genome Research. PMID- 11856865 TI - Proposal for a collaborative research project on mapping genes of Xenopus laevis. PMID- 11856866 TI - Insertion of a loxP site in a size-reduced human accessory chromosome. AB - The generation in vitro of mammalian artificial chromosomes, in view of the possibility of developing new technologies for gene therapy, is still an ambitious goal. Mammalian artificial chromosomes, to be used as cloning and expression vectors, have been constructed either by de novo synthesis or by reduction of pre-existing chromosomes. In the work here reported, we introduced a loxP sequence into the pericentromeric region of a chromosome 9-derived X-ray reduced minichromosome, with the purpose of generating a human chromosome vector (HCV). The modified accessory chromosome is linear and mitotically stable, has lost at least 1400 kb of alpha satellite DNA and normally binds CENP-B, CENP-C and CENP-E. The efficiency of gene targeting via loxP mediated homologous recombination was tested using the histone H2B-Green Fluorescent Protein chimaeric gene as a reporter. The frequency of site-specific insertion of the exogenous sequence was found to be about 50% and to occur in a controlled way with regard to the number of copies. The expression level of the fusion protein was stable over prolonged time in culture. PMID- 11856867 TI - Fibroblast growth factor receptor 2 (FGFR2): genomic sequence and variations. AB - Fibroblast growth factor receptors (FGFRs) play an important role in development and tumorigenesis. Mutations in FGFR2 cause more than five craniosynostosis syndromes. The FGFR2 genomic structure is the largest of the FGFR family. We have refined and extended the genomic organization of the FGFR2 gene by sequencing more than 119 kb of PACs, cosmids, and PCR products and assembling a region of approximately 175 kb. Although the gene structure has been reported to include only 20 exons, we have verified the presence of at least 22 exons, some of which are alternatively spliced. The sizes of six exons differed from those reported previously. Comparison of our sequence and those in the NCBI database detected more than 300 potential single nucleotide polymorphisms (SNPs). However, sequencing regions containing 52 of these potential SNPs verified only 14 in PCR products generated from 16 CEPH alleles. In contrast, direct sequencing of the CEPH DNAs revealed 21 other polymorphisms. Only one SNP was found in the 2,926 bp of coding sequence. Twenty-seven SNPs, two insertion polymorphisms and five microsatellite polymorphisms are contained in approximately 16.6 kb of non-coding sequence. These data yield an average of one polymorphism for approximately 488 bp of non-coding sequence examined. This collection of SNP, insertion, and repeat polymorphisms will aid future association studies between the FGFR2 gene and human disease and will enhance mutation detection. PMID- 11856868 TI - Assignment to chromosome 12q24.33, gene organization and splicing of the human KRAB/FPB containing zinc finger gene ZNF84. AB - FISH analysis was used to assign the human ZNF84 gene to chromosome 12q24.33, a region associated with recurrent breakpoints and allelic loss in several human cancers. In this report we show that the ZNF84 coding region is organized in four exons; two are dedicated to encoding the KRAB/FPB-A and KRAB/FPB-B modules, the remaining exons encode the N-terminal amino acids and C-terminal array of zinc finger units, respectively. PMID- 11856869 TI - PHF3 expression is frequently reduced in glioma. AB - Glioblastoma is the most frequent brain tumor and accounts for approximately 50- 60% of all astrocytic tumors. Many chromosome alterations have been described in glioblastoma, but only for a few alterations were the genes identified and linked to genetic pathways in glioblastoma development. To contribute to the identification of novel genes involved in glioblastoma development we used a combined immunological and molecular screening approach. Here we report the identification and expression analysis of a novel gene from human chromosome 6q12 that is considered to be the third member of a family of PHD finger containing genes and is termed PHF3. PHF3 is ubiquitously expressed in normal tissues including brain, but its expression is significantly reduced or lost in glioblastoma, glioblastoma cell lines, anaplastic astrocytomas and astrocytomas. The PHF3 protein sequence contains several protein motifs frequently found in transcription factors. One of those motifs is a PHD finger, also termed LAP motif and known to bind large portions of DNA. Another region of the protein revealed a high homology to the transcription factor TFIIS, especially to a region that is necessary for the Polymerase II binding properties of TFIIS. Combining these results, PHF3 is a novel member of a large class of regulatory proteins containing a LAP motif, and loss of its expression in glioblastoma may contribute to glioma development. PMID- 11856870 TI - The pericentromeric region of human chromosome 11: evidence for a chromosome specific duplication. AB - We have identified a chromosome duplication in the pericentromeric region of human chromosome 11 located in 11p11 and 11q14. A detailed physical map of each duplicated region was generated to describe the nature of the duplication, the involvement at the centromere and to resolve the correct maps. All clones were evaluated to ensure they were representative of their genetic origin. The order of clones, based on their marker content, as well as the distance covered was determined by SEGMAP. Each duplication encompasses more than 1 Mb of DNA and appears to be chromosome 11 specific. Ten STS markers were mapped within each duplication. Comparative sequence analysis along the duplication identified 35 nucleotide changes in 2,036 bp between the two copies, suggesting the duplication occurred over 14 million years ago. A suggested organization of the pericentromeric region, including the duplications and alpha-related repetitive sequences, is presented. PMID- 11856872 TI - Genomic structures of the human angiopoietins show polymorphism in angiopoietin 2. AB - The family of human angiopoietins comprises factors with important roles in vascular development and angiogenesis. All angiopoietins bind with similar affinity to the endothelial cell-specific receptor, Tie2. The mechanism by which they contribute to angiogenesis is thought to involve regulation of endothelial cell interactions with supporting perivascular cells. In this study the genomic structures of all three human angiopoietins were characterised by direct sequencing of human genomic DNA from the appropriate P1 artificial chromosome (PAC) clones. The exact positions at the intron/exon boundaries and the lengths of all eight introns were determined. As would be expected from the homology of these three proteins, the positions of the introns in the three genes are highly conserved. The putative RNA transcription start site for each angiopoietin gene was also determined. Intron-specific primers were used to amplify each exon of angiopoietin-1 and angiopoietin-2 from individual genomic DNAs. Although no polymorphism has been detected in the coding region of angiopoietin-1, three independent polymorphisms have been identified for angiopoietin-2. PMID- 11856873 TI - Down syndrome cell adhesion molecule is conserved in mouse and highly expressed in the adult mouse brain. AB - Down Syndrome (DS) is a major cause of mental retardation and is associated with characteristic well-defined although subtle brain abnormalities, many of which arise after birth, with particular defects in the cortex, hippocampus and cerebellum. The neural cell adhesion molecule DSCAM (Down syndrome cell adhesion molecule) maps to 21q22.2-->q22.3, a region associated with DS mental retardation, and is expressed largely in the neurons of the central and peripheral nervous systems during development. In order to evaluate the contribution of DSCAM to postnatal morphogenetic and cognitive processes, we have analyzed the expression of the mouse DSCAM homolog, Dscam, in the adult mouse brain from 1 through 21 months of age. We have found that Dscam is widely expressed in the brain throughout adult life, with strongest levels in the cortex, the mitral and granular layers of the olfactory bulb, the granule cells of the dentate gyrus and the pyramidal cells of the CA1, CA2 and CA3 regions, the ventroposterior lateral nuclei of the thalamus, and in the Purkinje cells of the cerebellum. Dscam is also expressed ventrally in the adult spinal cord. Given the homology of DSCAM to cell adhesion molecules involved in development and synaptic plasticity, and its demonstrated role in axon guidance, we propose that DSCAM overexpression contributes not only to the structural defects seen in these regions of the DS brain, but also to the defects of learning and memory seen in adults with DS. PMID- 11856871 TI - Molecular and cytogenetic characterisation of a small interstitial de novo 20p13- >p12.3 deletion in a patient with severe growth deficit. AB - We report on a small de novo interstitial deletion of the short arm of chromosome 20, 46,XY,del(20)(p12.3p13), in a young boy with hypotonia, moderate development delay, mild facial dysmorphism and severe growth failure. This patient did not show major features of Alagille-Watson Syndrome (AWS) which are common in more proximal 20p deletions. Standard and high resolution chromosome banding analysis revealed an apparent terminal deletion. Nevertheless, using chromosomal fluorescent in situ hybridization (FISH) and molecular analysis with polymorphic markers, we demonstrated that the abnormal chromosome resulted from a de novo interstitial deletion of paternal origin spanning from D20S842 to D20S900 and covering approximately 6 Mb. These findings indicate that a karyotype can lead to insufficient characterization of an apparently terminal deletion, and that one or a few genes in 20p13-->p12.3 bands are important for normal growth. PMID- 11856874 TI - Isolation of novel mouse genes associated with ectopic ossification by differential display method using ttw, a mouse model for ectopic ossification. AB - Mouse mutant ttw (tiptoe walking) is an excellent model for ectopic ossification. This mutant exhibits ossification in various soft tissues, which is histologically similar to human OPLL (ossification of posterior longitudinal ligament of the spine). We previously reported that ttw is caused by a nonsense mutation of the nucleotide pyrophosphatase (ENPP1) gene, and that a polymorphism of the human ENPP1 gene is associated with OPLL. These facts indicate that ENPP1 regulates ectopic ossification in vivo; however, the mechanism is unclear. ENPP1 is an ectoenzyme that generates phosphate (Pi) and pyrophosphate (PPi). PPi is a strong inhibitor of ossification. Abnormal Pi metabolism is observed in patients with OPLL, and diseases with abnormal Pi metabolism such as hypophosphatemic rickets are frequently complicated by ectopic ossification. These lines of evidence suggest Pi-PPi metabolism associated with ENPP1 may play an important role in regulation of ectopic ossification. To clarify the molecular mechanism of ectopic ossification in ttw, we examined the effect of dietary phosphate and calcium on the ttw phenotype and found a high dietary phosphate-accelerated ectopic ossification. Then we examined genes associated with the enhanced ossification in ttw on a high phosphate diet by a differential display method. We identified nine mouse genes; six genes were up-regulated by the high phosphate diet, and three were down-regulated. Six of the nine genes were novel and we cloned and characterized them. Two of the genes were highly specific to cartilage, suggesting their specific role in enchondral ossification. Our identification of the novel genes would give novel insight into the mechanism of ectopic ossification and etiology of OPLL. PMID- 11856875 TI - Syrian hamster 5S rRNA genes are assigned to 6q2 with PNA-FISH by an R-banded karyotype. AB - A karyotype for the Syrian hamster is proposed based on an R-banding pattern. R bands were obtained by BrdU incorporation into the cells followed by a combined DAPI and propidium iodide staining of the fixed metaphase spreads. In situ hybridisation was performed with two biotinylated 18-mer PNA (peptide nucleic acid) probes complementary to sequences within the 5S rRNA gene. The 5S rRNA gene repeats map to chromosome 6q2. The present PNA-FISH procedure is an abbreviated and simpler version of that previously published. PMID- 11856876 TI - Comparative mapping of the human 9q34 region in Fugu rubripes. AB - Twenty-seven genes have been cloned and mapped in Fugu which have orthologues within the human chromosome 9q34 region. The genes are arranged into five cosmid and BAC contigs which physically map to two different Fugu chromosomes. Considering the gene content of these contigs, it is more probable that a translocation event took place early in the Fugu lineage to split the ancestral 9q34 region onto two chromosomes rather than the alternative hypothesis of a large-scale duplication of the region into two chromosomes with subsequent rapid and dramatic gene loss. There are considerable differences in gene order between the two species, which would appear to be the result of a series of complex chromosome inversions; thus suggesting that there have been no positional constraints on this particular gene set. PMID- 11856877 TI - Comparative FISH mapping of 32 loci reveals new homologous regions between donkey and horse karyotypes. AB - A total of 32 loci comprising specific genes, microsatellites and anonymous BAC clones from horse and cattle were mapped on donkey chromosomes. Of these, 13 markers were also mapped for the first time in the horse. This information, together with that previously available in donkey and horse updates the comparative status of the karyotypes of the two species. The findings of the present study for the first time show correlation between eleven equine acrocentric autosomes and the donkey chromosomes and in part enable detection of rearrangements between them. There are still 7-8 pairs of chromosomes/arms for which no correspondence is known. At least 20 chromosome rearrangements (inversions, fusions and fissions) are already identified that differentiate the two karyotypes. More will be known once complete correspondence is deduced between them. These observations match similar differences observed between human gibbon and mouse-rat karyotypes that show considerable rearrangements in relation to each other. How donkey and horse karyotypes gathered these differences within a short period of 5-10 Myr since divergence from a common ancestor will be known only after an ancestral equid karyotype is deduced, and the direction of change leading to chromosome rearrangements is clearly understood. PMID- 11856878 TI - Characterization and chromosome assignment of the canine gamma-sarcoglycan gene (SGCG) to CFA 25q21-->q23. AB - Mutations in the gene for gamma-sarcoglycan (SGCG) located on HSA 13q12 are responsible for limb girdle muscular dystrophy (LGMD2C) in human. Here we report the cloning of the canine SGCG gene together with its genomic structure and several intragenic polymorphisms. The coding part of the canine SGCG contains seven exons spanning at least 70 kb of genomic DNA. The chromosome assignment of the canine SGCG gene to CFA 25q21-->q23 confirms that the canine syntenic group 10 corresponds to CFA 25 and also supports the findings of human-canine reciprocal chromosome painting. PMID- 11856879 TI - Molecular characterization and chromosome assignment of the porcine gene COX7A1 coding for the muscle specific cytochrome c oxidase subunit VIIa-M. AB - The COX7A1 gene encodes a heart- and muscle-specific isoform of the subunit VIIA of cytochrome c oxidase, which is the last component of the mitochondrial electron transfer chain. Cloning and characterization of the porcine COX7A1 gene revealed a highly conserved organization with respect to other mammalian COX7A1 orthologs. The porcine gene consists of four exons spanning approximately 1.5 kb and codes for a peptide of 80 amino acids. The COX7A1 gene showed no variation between pigs from different breeds. The gene was assigned by FISH and RH-mapping to SSC 6q1.1-->q1.2 which is in agreement with previously established comparative maps. PMID- 11856880 TI - The canine ERBB2 gene maps to a chromosome region frequently affected by aberrations in tumors of the dog (Canis familiaris). AB - The dog offers an increasingly important model for several human diseases, including cancer. Accordingly, the results of canine gene mapping studies will be of considerable significance. Herein, we have addressed the mapping of the canine gene ERBB2 (alias HER2, NEU). ERBB2 is a protooncogene encoding a tyrosine kinase receptor protein, the overexpression of which correlates with a more rapid progression and a worse prognosis in breast cancer. In addition, it apparently plays a role in the development of other tumors as well. By fluorescence in situ hybridization (FISH), we have mapped the canine ERBB2 to 1q13.1. Cytogenetic studies of canine tumors revealed that this region is very often affected by clonal chromosome aberrations in tumors of the dog. PMID- 11856881 TI - Human and mouse orthologs of a new ATP-binding cassette gene, ABCG4. AB - We characterized a new ATP-binding cassette (ABC) transporter gene from human and mouse that is highly expressed in the brain. The gene, ABCG4, produces several transcripts that differ at the 5' end and encode proteins of various lengths. The ABCG4 protein is closely related to the Drosophila white and human ABCG1 genes, and belongs to the ABCG subfamily several members of which are involved in cholesterol transport. All representatives of this "reverse transporter" subfamily, including ABCG4, have a single ATP-binding domain at the N-terminus and a single C-terminal set of transmembrane segments. ABCG4 maps to human chromosome 11q23, between the markers D11S939 and D11S924, and Abcg4 to a conserved syntenic region on mouse chromosome 9. The abundant expression of this gene in the brain and close evolutionary relationship to the other members of the subfamily suggests a potential role for ABCG4 in cholesterol transport processes in this tissue. PMID- 11856882 TI - A pericentric inversion in the cattle Y chromosome. AB - Sixteen male Podolian cattle, two sires and their 14 male offspring, were investigated cytogenetically on the basis of a female-like phenotype found in one of them. Eleven male offspring, including the one with female traits, and one of the two sires were found to carry an abnormal Y chromosome which originated from a pericentric inversion of the proximal half of the Yq arm (Yq11-->q12.2), as demonstrated by both banding and FISH mapping techniques with Y-specific molecular markers. PMID- 11856883 TI - Molecular cytotaxonomy of New World monkeys (Platyrrhini) - comparative analysis of five species by multi-color chromosome painting gives evidence for a classification of Callimico goeldii within the family of Callitrichidae. AB - Chromosome rearrangements are considered as "rare genomic changes" and can provide useful markers and even landmarks for reconstructing phylogenies complementary to DNA sequence data and bio-morphological comparisons. Here, we applied multi-directional chromosome painting to reconstruct the chromosome phylogeny and evolutionary relationships among the New World monkey (Platyrrhini) species Callithrix argentata, Cebuella pygmaea, Saguinus oedipus, Callithrix jacchus and Callimico goeldii. The results clarified several aspects of New Wold monkey phylogeny. In particular the phylogenetic position of C. goeldii was elucidated, which has been controversially discussed and variously classified in the family Callitrichidae, in the family Cebidae or in its own family Callimiconidae. Comparative genome maps were established by multi-color fluorescence in situ hybridization (FISH) with human, S. oedipus and Lagothrix lagothricha chromosome- specific DNA probes. From these data we reconstructed the putative ancestral karyotype of all Callitrichidae. Various derived chromosomal syntenies are shared by all five species and cytogenetically define Callitrichidae - including Callimico goeldii -- as a distinctive group within the Platyrrhini. C. pygmaea and C. argentata share identical chromosomal syntenies from which S. oedipus and C. jacchus differ by single independent translocations. A common derived chromosomal change links Callimico with the marmosets to the exclusion of the tamarins, however, it has further diverged from an ancestral marmoset karyotype by at least four apomorphic rearrangements. Saimiri sciureus, representing the Cebinae, exclusively shares a derived syntenic association with all Callithrichidae, defining the genus Saimiri as a sister group. PMID- 11856884 TI - Identification and characterization of alternatively spliced murine Rgs11 isoforms: genomic structure and gene analysis. AB - The RGS proteins comprise a large family of proteins which were recently identified as negative Regulators of G-protein Signaling. They have been shown to act as GTPase Activating Proteins (GAPs) towards the G(alpha) subunits of heterotrimeric G-proteins. In addition to this GAP activity, which has been shown to occur through the RGS domain, RGS proteins are likely to possess other functions due to the existence of other domains in these molecules (De Vries and Farquhar, 1999; Hepler, 1999). Here, we report the molecular characterization of the murine Rgs11 gene. The gene encodes a protein with high homology to human RGS11 (79.9%), containing conserved DEP (Dishevelled/EGL-10/Pleckstrin) and GGL (G protein gamma-like) domains. The gene is comprised of at least 13 exons, spanning 8-9 kb. Spliced transcript variants were identified which are co expressed with 5A3, a transcript that contains the largest ORF. Expression of mouse Rgs11 was found to be restricted to specific tissues with a unique pattern of expression observed in brain. PMID- 11856885 TI - A new balanced autosomal reciprocal translocation in cattle revealed by banding techniques and human-painting probes. AB - Three hundred and twenty-two (264 males and 58 females), randomly sampled Grey Alpine cattle individuals from Northeastern Italy, were investigated cytogenetically by both conventional chromosome staining and R-banding. Two hundred and eighty-one (87%) individuals had a normal karyotype and 41 (13%) carried chromosomal aberrations such as (a) rob(1;29) in two individuals, (b) rob(26;29) in 36 individuals, (c) XX/XY-chimerism in two individuals, and (d) an abnormally long chromosome in one individual. All these aberrations except (d) have been described before. GBG-, RBG-, CBA-banding and sequential GBG/CBA- and RBG/CBA-banding techniques revealed that the abnormally long chromosome was the result of a reciprocal translocation between chromosomes 1 (q21-->qter) and 5 (q11-->q33), as confirmed also by chromosome painting with human chromosome 3 and 12 probes. The dam of the carrier bull carried the same translocation, while the grandam showed a normal karyotype. Since the sire of the dam was not available for study, no conclusion about the origin of the chromosome translocation could be drawn. The carrier bull was eliminated because of poor fertility. The dam had three other calves, which all were chromosomally normal. On average the dam had to be served 2.5 times (breed average was 1.2) to be in calf. PMID- 11856886 TI - Alignment of VIM, MRC1, GAD2, and IL2RA genes on swine chromosome 10q by in situ hybridization and RH mapping. AB - Since the distal half of swine chromosome (SSC) 10q was shown to contain a quantitative trait locus (QTL) influencing swine growth, the precise correspondence between this chromosome region and the orthologous human chromosome region (HSA10p) was investigated using chromosomal fluorescence in situ hybridization and RH mapping of type I loci spanning the growth QTL. The goal was to align this critical region of swine with the corresponding region in human for the purpose of identifying candidate genes. The HSA10p type I loci mapped in swine were VIM, MRC1, GAD2, and IL2RA. Locus order on SSC10q was shown to be centromere-VIM-MRC1-GAD2-IL2RA, while in human the order is centromere-GAD2 MRC1-VIM--IL2RA, indicating that the chromosome segment marked by VIM, MRC1 and GAD2 has been inverted relative to the centromere and IL2RA. PMID- 11856887 TI - Expression pattern and further characterization of human MAGED2 and identification of rodent orthologues. AB - In a search for genes involved in X-linked mental retardation we have analyzed the expression pattern and genomic structure of human MAGED2. This gene is a member of a new defined MAGE-D cluster in Xp11.2, a hot spot for X-linked mental retardation. Rat and mouse orthologues have been isolated. In contrast to the genes of the MAGE-A, MAGE- B and MAGE-C clusters, MAGED2 is expressed ubiquitously. High expression was detected in specific brain regions and in the interstitium of testes. Five SNPs in the coding region of human MAGED2 were characterized and their allele frequencies determined in a German and Turkish population. PMID- 11856888 TI - Aneuploidy in human oocytes: nondisjunction or predivision? PMID- 11856889 TI - Assignment of Nmp4 to mouse chromosome 6 band F1 flanked by D6Mit134 and D6Mit255 using radiation hybrid mapping and fluorescence in situ hybridization. PMID- 11856890 TI - Assignment of the metastasis-associated gene (Mta1) to mouse chromosome band 12F and the metastasis-associated gene 2 (Mta2) to mouse chromosome band 19B by fluorescence in situ hybridization. PMID- 11856891 TI - Assignment of the bovine runt-related transcription factor 1 gene (RUNX1) to bovine chromosome 23q21 by fluorescence in situ hybridization and radiation hybrid mapping. PMID- 11856892 TI - Assignment of the BRCA1-associated RING domain gene (Bard1) to rat chromosome 9q34 by in situ hybridization and radiation hybrid mapping. PMID- 11856893 TI - Assignment of OVCOV1 (alias CGI-15) to human chromosome 20 band q13.1-->q13.2 by fluorescent in situ hybridization. PMID- 11856894 TI - A multiple schedule model of limited access drinking in the cynomolgus macaque. AB - The present study reports on the development of a multiple schedule procedure of oral ethanol self-administration in cynomolgus macaques. Six adult cynomolgus macaques (four female, two male) were trained to self-administer ethanol and water under a 60 min, four-component multiple schedule of ethanol and water access with 1 g food pellets presented every 900 s (fixed-time 900 s). Water was available for the first and third 15 min components, ethanol in the second and fourth components. Total ethanol dose was stable at between 1-1.25 g/kg at ethanol concentration of 4%, 6% and 8%. Subsequently water was replaced with a sweetened drink (sugar-free Tang powder, General Foods). Ethanol and Tang were self-administered in similar volumes and both served as reinforcers compared with water. Acute pretreatment with 0.25 to 1.5 g/kg of intragastrically gavaged (i.g.) ethanol failed to alter ethanol or Tang self-administration significantly despite producing mean blood ethanol levels of up to 199 mg/dl when combined with self-administered ethanol. However, 1.0 g/kg i.g. ethanol administered for 15 consecutive days significantly decreased ethanol self-administration by 23%. The results suggest that ethanol self-administration under a multiple schedule is insensitive to alteration by acute ethanol pretreatment, but can be decreased by more prolonged chronic ethanol pretreatment. PMID- 11856895 TI - Evidence that mnesic states govern normal and disordered memory. AB - Although it is recognized that retrieval may be state-dependent, only recently has a paradigm been identified that allows state-dependence in rats to be demonstrated reliably and at relevant doses of CNS agents. In humans, the effects of scopolamine constitute a valuable model of disordered memory. We used this paradigm to analyze the effects of scopolamine on different memory processes. Rats treated with either saline or 0.01-10 mg/kg doses of scopolamine learned to lever press for milk reward, and were then tested for retrieval while given the same or a different treatment. Saline-to-scopolamine as well as scopolamine-to saline state changes produced robust failures to retrieve the response. Remarkably, the state produced by 2.5 mg/kg scopolamine, like that produced by saline, produced little intrinsic effect on learning or any other memory process (i.e. when the prevailing state was left unchanged). However, changing the implemented state from one to the other between two different processes, disrupted not only retrieval, but also learning, encoding and retention. We also determined whether the graded state changes produced by 0.01-10 mg/kg doses of scopolamine could mimic the peculiar and poorly understood temporally graded retrograde amnesia that occurs in Alzheimer's disease. In rats that had acquired a complex drug-discrimination task over a 6-month period, scopolamine-induced state changes seemed to produce dose-dependent deficits in the retrieval of recent information while preserving those abilities that had been acquired in the increasingly remote past. Beyond its role in retrieval, the findings implicate state dependence in learning, encoding and retention, and suggest that physiologically defined mnesic states govern each of these. The changes of mnesic state that are likely associated with excessively labile cholinergic neurotransmission may conceivably cause the complex disabilities of Alzheimer's disease. PMID- 11856897 TI - Differential effects of dopamine antagonists on locomotor activity, conditioned activity and conditioned place preference induced by cocaine in rats. AB - Neuronal substrates that mediate the conditioned effects of cocaine have not been well characterized. To examine dopaminergic mechanisms, three antagonists were tested for their capacity to inhibit the expression of conditioned locomotor activity and conditioned place preference in rats. Antagonists were also assessed against acute cocaine-stimulated locomotor activity for comparison. For locomotor activity conditioning, six conditioning sessions were conducted over a 10-day period. Paired rats received 10 mg/kg cocaine prior to activity sessions and saline after; unpaired controls received saline prior and cocaine after. For place preference conditioning, eight conditioning sessions were conducted over a 13-day period; rats received 10 mg/kg cocaine while restricted to one of two distinct chambers and, on alternate days, they received saline in the other. Antagonists (haloperidol, raclopride and SCH23390; 0.03-0.1 mg/kg) were given only on test days for conditioned effects. All three antagonists significantly and dose-dependently attenuated the direct stimulatory effect of cocaine. SCH23390 showed a tendency to reduce the expression of conditioned locomotor activity, and only haloperidol blocked the expression of conditioned place preference. Thus, direct and conditioned stimulant effects of cocaine were shown to be differentially sensitive to dopamine receptor blockade. Further, conditioned stimulant effects differed from conditioned reinforcing effects in this regard. PMID- 11856896 TI - Receptor-selective antagonism of opioid antinociception in female versus male rats. AB - This study was conducted to determine whether sex differences in opioid antinociception may be explained by sex differences in opioid receptor activation. The time course, dose-effect and selectivity of antagonists that have been previously shown to be relatively mu (beta-funaltrexamine, beta-FNA), kappa (norbinaltorphimine, norBNI), or delta (naltrindole, NTI) receptor selective in male animals were compared in female and male Sprague-Dawley rats using a 52 degrees C hotplate test. In both sexes, beta-FNA (10 or 20 microg intracerebroventricularly [i.c.v.]) dose-dependently blocked the antinociceptive effects of fentanyl (0.056 mg/kg subcutaneously); antagonism was observed 24 h after beta-FNA, and diminished within 7-14 days. In both sexes, norBNI (1 or 10 microg i.c.v.) dose-dependently blocked the antinociceptive effects of U69,593 (1.0 mg/kg subcutaneously); antagonism was maximal by 1-3 days post-norBNI and lasted longer than 56 days. NTI (1 or 10 microg i.c.v.) dose-dependently blocked the antinociceptive effects of [D-Pen2, D-Pen5]enkephalin (DPDPE, 100 nmol i.c.v.) in both sexes; however, the duration of action of NTI was shorter in females than in males. The antinociceptive effects of the mu receptor-preferring agonists fentanyl, morphine and buprenorphine were significantly and dose dependently antagonized by beta-FNA, but not by norBNI or NTI, in both sexes. Beta-FNA antagonism was significantly greater in females compared with males given morphine, but not fentanyl or buprenorphine. The antinociceptive effects of the kappa receptor-preferring agonists U69,593 and U50,488 were significantly and dose-dependently antagonized by norBNI; U50,488 but not U69,593 was also antagonized to a lesser extent by NTI and beta-FNA, in both sexes. The antinociceptive effect of the delta receptor-preferring agonist SNC 80 was significantly antagonized by NTI, but not by norBNI or beta-FNA, in both sexes. The sex difference in beta-FNA antagonism of morphine suggests that there may be sex differences in functional mu opioid receptor reserve or signal transduction; however, the lack of consistency across all mu agonists weakens this hypothesis. Overall, the opioids tested had very similar receptor selectivity in male and female subjects. PMID- 11856899 TI - Four-choice drug discrimination in pigeons. AB - (+)Amphetamine was added as a training stimulus for pigeons previously trained to discriminate among pentobarbital, morphine and saline using a three-choice procedure. Pigeons quickly learned the four-choice drug discrimination. Generalization from the training drugs was similar to that established with simpler drug discriminations; pentobarbital generalized to chlordiazepoxide, morphine generalized to methadone, and (+)amphetamine generalized to cocaine and methamphetamine. Low doses of phencyclidine generalized to saline, while higher doses partially generalized to pentobarbital and (+)amphetamine. When dose response curves were redetermined with a cumulative-dosing procedure, the same pattern of generalization occurred as under single-dose procedures. Dose-response curves were quantal under both the single-dose and the cumulative-dosing procedures. The four-choice procedure offers some important advantages for studying the discriminative stimulus effects of drugs that interact with multiple receptor subtypes and for studying drug mixtures. PMID- 11856898 TI - m-CPP hypolocomotion is selectively antagonized by compounds with high affinity for 5-HT(2C) receptors but not 5-HT(2A) or 5-HT(2B) receptors. AB - The ability of m-CPP [1-(m-chlorophenyl)piperazine] to produce hypolocomotion is well documented. This effect has been postulated to be due to activation of the 5 HT(2C) receptor. It is only recently that the tools necessary to clearly delineate which serotonin receptors are involved in the mediation of m-CPP hypolocomotion have become available. We investigated the effects of the selective 5-HT(2A) antagonists, MDL 100,907 and ketanserin, the selective 5 HT(2B) antagonists, LY 202146 and LY 266097, the 5-HT(2B/2C) antagonist, SB 206553, and the selective 5-HT(2C) antagonist, SB 242084 on m-CPP-induced hypolocomotion and spontaneous locomotor activity in mice. Furthermore, we investigated the effects of the non-selective serotonin antagonists, ritanserin, LY 53857, mianserin and cyproheptadine on m-CPP hypolocomotion. Additionally, receptor-binding studies were employed as an in vitro assessment of relative affinities at the 5-HT(2A), 5-HT92B) and 5-HT(2C) receptors. Antagonists tested alone were without effect on spontaneous activity, with the sole exception of ketanserin, which decreased spontaneous activity at the high dose of 1 mg/kg. m CPP-induced hypolocomotion was not significantly attenuated by various doses of MDL 100,907, ketanserin, LY 202146, LY 266097, ritanserin or cyproheptadine. In contrast, SB 206553, SB 242084, LY 53857 and mianserin were capable of reversing m-CPP-induced hypolocomotion. Consistent with previous suggestions, a detailed pharmacological evaluation with selective antagonists for the 5-HT2 family of receptors supports a primary role for the 5-HT(2C) receptor, and not 5-HT(2A) or 5-HT(2B) receptors, in mediating the hypolocomotion produced by m-CPP. PMID- 11856900 TI - LY294002, an inhibitor of phosphoinositide 3-kinase given into rat hippocampus impairs acquisition, consolidation and retrieval of memory for one-trial step down inhibitory avoidance. AB - Adult male Wistar rats were bilaterally implanted with indwelling cannulae in the CA1 region of the dorsal hippocampus. Once recovered from surgery, animals were submitted to one session of step-down inhibitory avoidance training (3.0 s, 0.4 mA footshock). Animals received a 0.5-microl infusion of saline, or of LY294002 (5, 50 or 500 microM), an inhibitor of the phosphoinositide 3-kinase (PI 3-K) family. Infusions were given 10 min before training, immediately post-training or 10 min prior to a 24-h retention test. In the pre- and post-training groups, the animals were tested twice: at 1.5 and 24 h after training, for short- (STM) and long-term memory (LTM), respectively. Pre- and post-training infusion of the drug inhibited both STM and LTM. Pre-test infusions impaired LTM retrieval. The effects can not be attributed to influences on locomotor, exploratory, pro- or anti-conflict behaviour, since LY294002 had no influence on elevated plus-maze behaviour. The results suggest that hippocampal PI 3-K is necessary for memory acquisition, consolidation and retrieval of the consolidation of step-down inhibitory avoidance in rats. This could be due to an interaction with the N methyl-d-aspartate (NMDA) receptor complex or with activity of the extracellularly regulated protein kinase (ERK)-Ras signalling pathway. PMID- 11856901 TI - A preliminary investigation on the acute pharmacodynamic effects of hypericum on cognitive and psychomotor performance. AB - Research has indicated that the herb St John's Wort (Hypericum perforatum) has comparable efficacy to conventional antidepressants in the treatment of depression. Although clinical studies have demonstrated that hypericum has a superior side-effect profile compared to standard antidepressants, no study has directly compared the cognitive and psychomotor effects of hypericum with those of other antidepressants. The aim of the current study was to examine the acute effects of hypericum on cognitive and psychomotor function, and to compare its effects with those of amitriptyline. Thirteen healthy volunteers received an acute dose of placebo, amitriptyline (25 mg, positive control) or hypericum (900 mg or 1800 mg) in a double-blind, placebo-controlled design. Cognitive and psychomotor tests and subjective measures of sedation were administered before and 1, 2 and 4 hours after drug administration. Amitriptyline impaired performance on a battery of psychological tests, which included critical flicker fusion (CFF), choice reaction time (CRT), digit symbol substitution test (DSST), profile of mood states (POMS) and the line analogue rating scale (LARS), while hypericum had neutral effects on performance in these tests. However, hypericum induced a dose-related impairment on DSST. Current findings suggest that clinical doses of hypericum do not impair attention, sensorimotor function or information processing. PMID- 11856902 TI - Activity of SR141716 on post-reinforcement pauses in operant responding for sucrose reward in rats. AB - Cannabinoids increase food intake, via CB1 receptors. The CB1 antagonist, SR141716, has been reported to decrease palatable food consumption in both operant and non-operant procedures. Similarly, CB1 receptor blockade diminished responding for normal food pellets under a fixed-ratio 15 (FR-15) schedule of reinforcement. The present experiment investigated whether the control of a continuous schedule of reinforcement (CRF) for sucrose pellets would be sensitive to the CB1 antagonist in mildly deprived rats. SR141716 dose-dependently reduced responding in a CRF procedure, by increasing post-reinforcement pauses. Together with formerly published conclusions, the data suggest that CB1 blockade reduces the rewarding efficacy of both palatable and non-palatable food. PMID- 11856903 TI - 3-Hydroxy-3-methylglutaryl coenzyme A reductase inhibitors and rhabdomyolysis: considerations in the renal failure patient. AB - An intense debate has developed as to the risk-benefit ratio of 3-hydroxy-3 methylglutaryl coenzyme A reductase inhibitors (statins) following the withdrawal of cerivastatin. The development of rhabdomyolysis in cerivastatin-treated patients should have surprised few since myotoxicity is an accepted class effect of statins. What has sprung from the cerivastatin experience though is a concern for other members of this class. Such misgivings, although understandable, are ill advised. Without question, differences exist in the risk of rhabdomyolysis occurrence amongst the various statins. In this regard, pravastatin and fluvastatin are least likely to produce rhabdomyolysis, which, in part, relates to the fact they are not metabolized by the cytochrome P450 3A4 pathway. When muscle damage occurs with statins it is most often the result of a drug-drug interaction rather than a specific adverse response to statin monotherapy. Such drug-drug interactions increase plasma concentrations of a statin and thereby increase the risk of myotoxicity. A growing consensus exists which supports an expanded use of statins in a range of patient groups including the renal failure patient. Polypharmacy and altered drug metabolism increase the risk of myotoxicity, albeit to an ill-defined degree, in this population. Many factors should enter into the choice of a statin in the multiply medicated renal failure patient. PMID- 11856904 TI - Optimal blood pressure control and antihypertensive regimens in hypertensive renal disease: the potential of exploring the mechanisms of response variability. AB - In renal patients rigorous blood pressure control is crucial to prevent renal and cardiovascular target organ damage. For renoprotection target blood pressure depends on the severity of proteinuria before treatment. For proteinuria of 1- 3g/day a mean arterial pressure of 98 mmHg provides additional benefit, whereas the target should be as low as 92 mmHg if proteinuria exceeds 3g/day. The antiproteinuric effect of antihypertensive intervention predicts renoprotection; it is therefore recommended that therapy should be titrated not only on blood pressure, but also on reduction of proteinuria. All currently available classes of antihypertensives can be used to reduce blood pressure in renal patients. Interventions based on blockade of the renin-angiotensin-aldosterone system have additional antiproteinuric, and thus renoprotective, potential. Large individual differences in therapeutic benefit are common, even for interventions of proven efficacy at group level. Studies applying different classes of drugs in the same patient (rotation schedules) demonstrate that individual factors are main determinants of therapy response. Exploration of the mechanisms underlying these patient factors is important to improve treatment outcome. Analysis of genetic determinants of therapy response has great potential in this respect. However, therapy response is a complex phenotype. Thus, careful study of gene-gene and gene-environment interactions will be needed in order to turn this type of knowledge into benefit for the patient. PMID- 11856905 TI - Accelerated atherosclerosis, dyslipidemia, and oxidative stress in end-stage renal disease. AB - Premature atherosclerosis is a major cause of morbidity and mortality in end stage renal disease patients. Dyslipidemia and increased oxidative stress contribute to premature atherogenesis in these patients. The dyslipidemia of end stage renal disease consists of both quantitative and qualitative abnormalities in serum lipoproteins. Qualitative changes include hypertriglyceridemia (increased remnant lipoproteins), low high-density lipoprotein-cholesterol, and increased lipoprotein (a). In addition to quantitative changes, lipoproteins in end-stage renal disease undergo compositional and qualitative changes that make them pro-atherogenic, such as various modifications of apolipoprotein B, including oxidation, and modification by advanced glycation end-products. The 3 hydroxy-3-methylglutaryl coenzyme A reductase inhibitors and low-dose fibrates could be effective therapies for lipid disorders. The best evidence for increased oxidative stress in end-stage renal disease is the demonstration of increased plasma F2-isoprostanes. Confirmation of the positive findings with high-dose alpha-tocopherol in the Secondary Prevention with Antioxidants of Cardiovascular Disease in End-stage Renal Disease Study is urgently needed. Clinical trials with statins and other drugs that improve dyslipidemia also need to be undertaken. These therapies could clearly lead to a reduction in cardiovascular morbidity and mortality in these patients. PMID- 11856906 TI - Pharmacological management of membranous nephropathy. AB - When steroids and immunosuppressive drugs were the only available pharmacological agents used to treat membranous nephropathy, nephrologists were polarized into two groups, those supporting therapy on the basis of the results achieved in controlled trials and those opposed to therapy who contended that the side effects of therapy were too severe to consider in a disease with a relatively benign course. These two groups are drawing closer as treatments with lesser side effects emerge. The demonstration that proteinuria accelerates progressive kidney failure in all renal diseases led to a major focus on control of proteinuria. Angiotensin-converting enzyme inhibitors, diuretics and angiotensin II receptor antagonists all play a role. Older methods of treatment that reduce proteinuria are being resurrected. A major development is the demonstration in a randomized study that cyclosporin A is effective in membranous nephropathy. Therefore, although there has been no major recent breakthrough or novel therapeutic agent used in membranous nephropathy, a range of new methods of controlling proteinuria provide some compromise between therapeutic enthusiasm and conservative management in this common disorder. PMID- 11856908 TI - Daily hemodialysis: an update. AB - The interest in daily hemodialysis in the form of short daily hemodialysis and nocturnal hemodialysis has continued to increase over the past few years. A significant number of publications support the evidence for improved outcomes in most of the study areas, including financial benefits. The increased direct cost of the provision of daily hemodialysis seems to be the main obstacle to the wider acceptance of these methods. Convincing data of improved outcomes and evidence of cost efficiency of daily hemodialysis are being generated. These data may increase the interest of the institutional payors as well as of the dialysis industry to create the proper infrastructure for the use of daily hemodialysis. These dialysis regimens promise to have a major impact on the management of end stage renal disease. PMID- 11856907 TI - Recent developments in toxic nephropathy. AB - Toxic nephropathy is an important cause of reversible renal injury if detected early. Renal damage can be due to several different mechanisms affecting different segments of the nephron, renal microvasculature or interstitium. Clinical signs may not be apparent in the early stages and assessment of renal function should include thorough evaluation of glomerular filtration rate, proximal and distal tubular function. A kidney biopsy may be indicated to establish the cause and effect relationship. The presence of comorbid conditions such as older age, diabetes mellitus, hypertension and congestive heart failure have a significant influence on the patient's ability to recover from the toxic effects. A significant degree of drug-induced renal toxicity is only acceptable if the causative agent is used for the curative treatment of an underlying disease but not if the aim is the palliative or supportive therapy. The decision to reduce the dose or to stop the toxic agent must be based on the ultimate goal of therapy and the patient's baseline health status. PMID- 11856909 TI - Good news for patients with type 2 diabetes: angiotensin receptor blocker treatment delays progression of diabetic nephropathy. PMID- 11856910 TI - What is new in endothelium-derived hyperpolarizing factors? AB - The chemical identification and functional characterization of endothelium derived hyperpolarizing factors varies depending on vascular size, vascular bed and species. Three major candidates are the epoxyeicosatrienoic acids, cytochrome P450 metabolites of arachidonic acid, potassium ion and hydrogen peroxide. Additionally, electrical coupling through myoendothelial gap junctions serves to conduct electrical changes from the endothelium to the smooth muscle and may mediate or propagate hyperpolarization. Endothelium-derived hyperpolarizing factors are important mediators of vascular relaxation most specifically in resistance sized arteries where they regulate tissue blood flow. The release of the factors is modulated by a number of influences including agonist stimulation, shear stress, estrogen and disease. This article reviews the latest studies concerning the characterization of endothelium-derived hyperpolarizing factors, the mechanisms of factor release and alterations of the factors. PMID- 11856911 TI - Nitric oxide synthase and hypertension. AB - Nitric oxide, the metabolic product of L-arginine by the enzyme nitric oxide synthase, plays a pivotal role in the regulation of vascular homeostasis. Its complex interaction with the autocrine and paracrine systems, particularly angiotensin II, modulates vasoconstriction and vasodilatation as well as the architectural remodeling of the vascular bed. The major vascular hormones known to be involved are angiotensin II and endothelin-1. Upregulation of endothelin-1, a potent molecule, appears to be a consequence of the nitric oxide-angiotensin II imbalance that contributes to end-organ injury. Increased oxidative stress, common to different diseases including diabetes mellitus and hypertension, is also a determinant player in the interaction between angiotensin II and nitric oxide. The influence of a relative malfunction of the nitric oxide system on the vascular tone and vascular structure, and the effects of hypertension on this system, are discussed. PMID- 11856912 TI - Peroxisome proliferator-activated receptor-gamma in the renal mesangium. AB - The peroxisome proliferator-activated receptors (PPARs) are nuclear receptors that are expressed in a variety of tissues, including the liver (PPARalpha), adipose tissue, vascular smooth muscle, the heart, skeletal muscle, and the kidney (PPARgamma). PPARdelta is expressed ubiquitously. The receptors function as transcription factors to regulate the expression of genes involved in lipid metabolism, cell growth and migration as well as insulin-mediated skeletal muscle glucose uptake. Although the mechanisms by which all these actions occur have not been completely worked out, ligands to these receptors function to improve lipid metabolism, insulin sensitivity, endothelial dysfunction and urinary albumin excretion in patients with diabetes. Thus PPARs appear to have enormous implications for the management of cardiovascular disease. PMID- 11856913 TI - Sympathetic nervous system and the kidney in hypertension. AB - Long-term control of arterial pressure has been attributed to the kidney by virtue of its ability to couple the regulation of blood volume to the maintenance of sodium and water balance by the mechanisms of pressure natriuresis and diuresis. In the presence of a defect in renal excretory function, hypertension arises as the consequence of the need for an increase in arterial pressure to offset the abnormal pressure natriuresis and diuresis mechanisms, and to maintain sodium and water balance. There is growing evidence that an important cause of the defect in renal excretory function in hypertension is an increase in renal sympathetic nerve activity (RSNA). First, increased RSNA is found in animal models of hypertension and hypertensive humans. Second, renal denervation prevents or alleviates hypertension in virtually all animal models of hypertension. Finally, increased RSNA results in reduced renal excretory function by virtue of effects on the renal vasculature, the tubules, and the juxtaglomerular granular cells. The increase in RSNA is of central nervous system origin, with one of the stimuli being the action of angiotensin II, probably of central origin. By acting on brain stem nuclei that are important in the control of peripheral sympathetic vasomotor tone (e.g. rostral ventrolateral medulla), angiotensin II increases the basal level of RSNA and impairs its arterial baroreflex regulation. Therefore, the renal sympathetic nerves may serve as the link between central sympathetic nervous system regulatory sites and the kidney in contributing to the renal excretory defect in the development of hypertension. PMID- 11856914 TI - Sleep apnoea and hypertension. AB - This review article provides an update on two major issues. First, the most recent evidence supporting the occurrence of an association between obstructive sleep apnoea syndrome, or more generally sleep-disordered breathing, and arterial hypertension in humans is summarized and discussed. This includes an evaluation of both cross-sectional and longitudinal studies. Second, new insights into the mechanisms responsible for the appearance of chronic hypertension in individuals suffering from recurrent nocturnal apnoeic episodes are provided, based both on experimental studies in animals and on clinical studies in humans. The relevance of these data for the clinical management of hypertensive patients with sleep disordered breathing, and the possibility of obtaining a reduction in blood pressure through the application of nasal continuous positive air pressure, is also addressed. PMID- 11856915 TI - Left ventricular hypertrophy in hypertension as a predictor of coronary events: relation to geometry. AB - The present review examines epidemiological evidence for a relation of left ventricular hypertrophy with coronary heart disease, and mechanisms that may represent pathophysiological links between left ventricular hypertrophy and coronary events. Left ventricular hypertrophy has been demonstrated to be a powerful predictor of coronary heart disease, and when geometry is concentric the relation is even stronger. In addition to its association with risk factors for atherosclerosis and mechanisms that precipitate acute heart attacks, left ventricular hypertrophy also directly predisposes to and aggravates clinical presentation of coronary heart disease through a number of biological mechanisms. These include the following: increase in oxygen requirement related to left ventricular geometry; coronary hypertension, with endothelial dysfunction and reduced coronary reserve; diastolic dysfunction; and structural remodelling of myocardium and vascular bed. Some of these alterations are also worsened by underlying coronary heart disease, and can potentially be maintained by loop mechanisms. A recognizable stage of abnormal coronary haemodynamics in the context of left ventricular hypertrophy is probably that at which coronary reserve is impaired in the absence of any other sign of heart disease; in many circumstances, this may occur early in the disease process. PMID- 11856916 TI - Hypertension and diabetes. AB - Hypertension is often associated clinically with diabetes either as part of the insulin resistance syndrome or as a manifestation of renal disease. Elevated systemic blood pressure accelerates the progression of both microvascular and macrovascular complications in diabetes. Agents that interrupt the renin angiotensin system confer renoprotection via a range of hemodynamic and nonhemodynamic mechanisms. Recent clinical trials confirm that these agents confer renoprotection in type 1 and type 2 diabetic patients with early or advanced renal disease. Hypertension also appears to accelerate vascular and cardiac abnormalities in diabetes, including increased atherosclerosis, arterial stiffness, left ventricular hypertrophy and diastolic dysfunction. A number of recently published and ongoing trials are exploring the role of aggressive antihypertensive treatment with a range of antihypertensive drugs in diabetic subjects at risk of or with macrovascular disease. PMID- 11856917 TI - Hormone replacement therapy and hypertension. AB - Hypertension, particularly systolic hypertension, affects postmenopausal women and is an important risk factor for cardiovascular disease in this group. In the last year, several papers have better defined the mechanisms by which hormone replacement therapy influences blood pressure in postmenopausal women, including effects on nitric oxide, angiotensin II and vascular stiffness (compliance). Currently, data concerning the influence of hormone replacement therapy on blood pressure in postmenopausal women are inconclusive because of the limitations of published studies. It is clear, however, that hypertension is not a contraindication to starting hormone replacement therapy. PMID- 11856918 TI - Long-term effects of femoral derotation osteotomies: an evaluation using three dimensional gait analysis. AB - The purpose of this study was to evaluate the long-term effects of the femoral derotation osteotomy (FDO) in the ambulatory patient with cerebral palsy (CP). The effectiveness of the FDOs were evaluated using three-dimensional gait analysis just before surgery (P0), 1 year after surgery (P1), and 5 years after surgery (P2). A total of 20 patients (27 sides) with CP were evaluated. Related physical examination and motion measures showed significant improvements at P1 that were maintained at P2. Mean maximum internal hip rotation at P0 of 77 degrees +/- 9 degrees decreased to 53 degrees +/- 8 degrees at P1 and was maintained at 58 degrees +/- 11 degrees at P2. Mean maximum external hip rotation at P0 of 21 degrees +/- 11 degrees increased to 35 degrees +/- 15 degrees at P1 and was maintained at 32 degrees +/- 13 degrees at P2. Mean femoral anteversion at P0 of 63 degrees +/- 9 degrees was reduced to 26 degrees +/- 15 degrees and was maintained at 31 degrees +/- 13 degrees at P2. During gait, mean hip rotation in stance at P0 of 20 degrees +/- 8 degrees was decreased to 2 degrees +/- 10 degrees at P1 and was maintained at 4 degrees +/- 5 degrees at P2. There were associated significant foot progression changes from an internal progression mean of 5 degrees +/- 17 degrees at P0 to -11 degrees +/- 17 degrees at P1 that were maintained at -12 degrees +/- 5 degrees at P2. The findings suggest that the FDO is a viable and lasting treatment option for the correction of anteversion and associated internal hip rotation during gait in children with CP. PMID- 11856919 TI - Development and use of a wrist fusion plate for children and adolescents. AB - Arthrodesis is a widely accepted treatment for several destructive disorders of the wrist joint. Previous literature has shown that compression plating is a reliable technique for achieving rigid fixation and fusion in adults. The goal of this study was to apply similar principles of adult wrist fusion to children, using a custom-designed fusion plate sized for children. Five children underwent wrist fusion using rigid fixation and dorsal plate application. Mean age at time of surgery was 16.4 years. The indication for surgery was paralysis (spinal cord injury) in three and spasticity (cerebral palsy and traumatic brain injury) in two. Surgery was performed through a dorsal approach and combined with carpectomy in the two patients with spasticity. Follow-up averaged 2.2 years and radiographs showed solid fusion and proper alignment of plate and screws. Improved stability and limb function were noted by patients and family. There were no instances of hardware failure, and plate removal has not been required. PMID- 11856921 TI - Intramuscular psoas lengthening improves dynamic hip function in children with cerebral palsy. AB - The efficacy of psoas surgery for the treatment of hip flexor dysfunction in cerebral palsy has long been a subject of debate. A retrospective, repeated measures analysis was performed to examine the effect of psoas surgery based on gait and clinical and functional measures. The results of this study show that intramuscular psoas lengthening over the pelvic brim is a safe and effective way to improve the hip function of independently ambulatory children with cerebral palsy while maintaining hip flexor power. The data also support the use of the multivariate hip flexor index as an overall measure of hip function. PMID- 11856920 TI - Multilevel surgery in spastic diplegia: evaluation by physical examination and gait analysis in 25 children. AB - Gait improvement surgery was performed on 25 ambulatory children with the diplegic type of cerebral palsy. Multiple soft tissue and bony procedures were performed (mean 8.2 procedures) according to criteria defined on the basis of physical examination and gait analysis. Relevant physical examination findings and kinematic and kinetic data in the sagittal plane were evaluated before surgery and at least 3 years after surgery. Physical examination showed a reduction in the ankle plantar-flexor power and in the range of hip flexion and ankle plantarflexion after surgery. Analysis of gait data showed significant improvements in the sagittal plane kinematics and the power generation at the hip and the ankle. At the knee joint there was maintenance of power of the flexor and extensor group of muscles on physical examination, with significant improvements in the kinematics after surgery. The authors conclude that well-selected surgery improves function of the spastic muscle. The importance of assessing clinical, kinematic, and kinetic data together for proper evaluation of gait is stressed. PMID- 11856922 TI - Technique for iliopsoas ultrasound-guided active electromyography-directed botulinum a toxin injection in cerebral palsy. AB - Symptomatic hip flexion deformity secondary to iliopsoas spasticity may interfere with gait, impair sitting balance, or contribute to hip subluxation or dislocation. A nonsurgical, minimally invasive technique to ameliorate iliopsoas spasticity is presented. The technique uses intramuscular injections of botulinum A toxin to provide selective neuromuscular blockade of the iliacus or psoas muscles or both. Because of the anatomic location of the target muscles, this technique uses ultrasound guidance for needle placement. Active electromyographic stimulation is used to verify the needle position adjacent to active myoneural interfaces. The authors' experience to date includes the treatment of 28 patients (53 hips). Use of this technique has resulted in improved hip range of motion. No intraoperative or postoperative adverse events or complications have been observed. PMID- 11856923 TI - Outcome of medial versus combined medial and lateral hamstring lengthening surgery in cerebral palsy. AB - Pre- and postoperative gait analysis and static measurements from 37 children with cerebral palsy who underwent hamstring lengthening were evaluated. Significant improvements in static and kinematic measures were noted after surgery in both groups. Although the differences were not statistically significant, there was a suggestion that combined medial/lateral hamstring lengthening may provide greater improvement in popliteal angle and maximum knee extension in stance. However, there also appears to be a greater risk of knee hyperextension during gait after combined medial and lateral hamstring lengthening than after medial hamstring lengthening alone. Postoperative calf spasticity also appears to be a risk factor for postoperative knee hyperextension. Assessment of calf spasticity may be important in patients undergoing medial and lateral hamstring lengthening. Additional treatments such as bracing and/or botulinum toxin injections to the calf to control equinus and knee hyperextension may be beneficial. PMID- 11856924 TI - Assessment of femoral anteversion in children with cerebral palsy: accuracy of the trochanteric prominence angle test. AB - Clinical assessment of femoral anteversion (FA) in children with cerebral palsy (CP) is frequently determined by the trochanteric prominence angle test (TPAT). Limited three-dimensional volumetric imaging by axial tomography of the femur was performed before surgery for 35 hips in 20 children with CP. The TPAT was performed before the imaging study for 31 hips in 18 children. The TPAT angle was within 10 degrees of the FA as determined from the computed tomography scans (Murphy technique) for 17 femurs (55%). The most prominent portion of the greater trochanter was located anterior to the femoral neck axis (mean 27 degrees, range 0 degrees-52 degrees) on the three-dimensional images in 34 of 35 hips. A simulated TPAT, measured from the imaging studies, consistently underestimated the FA as determined by the Murphy technique (mean 10 degrees, range 0 degrees-18 degrees). Accurate clinical assessment of FA by the TPAT in children with CP presumes that the prominence of the greater trochanter lies perpendicular to the axis of the femoral neck. Three-dimensional imaging showed the prominence to be anterior, to a variable degree, to the femoral neck axis, which in addition to clinical factors such as obesity compromises the accuracy of this clinical maneuver. PMID- 11856925 TI - Decreased bone density in ambulatory patients with duchenne muscular dystrophy. AB - Osteoporosis has been recognized in nonambulatory boys with Duchenne's muscular dystrophy. In this study, the hips and spines of 10 ambulatory children with Duchenne's muscular dystrophy were examined. The study group comprised 10 boys with a mean age of 8 years (range 6-11). All patients were scanned by dual-energy x-ray absorptiometry (DEXA) using a DPX LUNAR Corporation scanner (Madison, WI, U.S.A.). Manual muscle testing of the major muscle groups of the upper and lower extremities was performed on all patients. Pubertal development status was determined using the Tanner scale. Although they were still ambulatory, boys diagnosed with Duchenne's muscular dystrophy experienced weakness in the lower extremity musculature, as well as proximal femur and lumbar spine osteoporosis or osteopenia. PMID- 11856926 TI - Mangled extremity severity score in children. AB - Treatment of the severely traumatized or mangled lower extremity poses significant challenges. The Mangled Extremity Severity Score (MESS) is a scale that uses objective criteria to assist with acute management decisions. Most research on the MESS has been in adults or combined series with few children. The study was performed to investigate the MESS in children exclusively. The MESS was applied retrospectively to 36 patients with grades IIIB and IIIC open lower extremity fractures collected from two level 1 pediatric trauma centers. Patients were divided into limb salvage and primary amputation groups based on the decision of the treating surgeon. In the salvage group there were 18 grade IIIB fractures and 10 grade IIIC fractures. The MESS prediction was accurate in 93% of the injured limbs. In the amputation group eight limbs met the inclusion criteria; the MESS agreed with the treating surgeon in 63% of cases. These findings suggest the MESS should be considered when managing a child with severe lower extremity trauma. PMID- 11856927 TI - Position of immobilization for pediatric forearm fractures. AB - The purpose of this study was to evaluate the effect of forearm position on residual fracture angulation for pediatric distal-third forearm fractures at the time of union. One hundred nine pediatric distal-third forearm fractures undergoing closed reduction and casting were prospectively randomized to be immobilized in pronated, supinated, or neutral position. Initial angulation and displacements were radiographically compared with healed fracture angulation at a minimum of 6 weeks. With 99 complete patient files, 38 fractures were casted in neutral, 26 in pronated, and 35 in supinated positions. Average initial angulation was 20 degrees; postreduction angulation measured 3 degrees. Final angulation at union averaged 7 degrees for all fractures. Forearm position failed to show a significant effect on fracture angulation at union. Residual fracture angulation at the time of union for pediatric distal-third forearm fractures was not significantly affected by forearm position (pronation, supination, neutral) during cast immobilization. PMID- 11856928 TI - Trochlear deformity occurring after distal humeral fractures: magnetic resonance imaging and its natural progression. AB - Eighteen children (mean age 50.7 months) with trochlear deformities occurring after and related to distal humeral fractures (12 Salter-Harris type 2, 5 supracondylar, 1 Salter-Harris type 4) were evaluated with serial radiographs. The carrying angle and range of motion were measured throughout the follow-up period (up to 13 years after trauma) after definitive treatment. Bony defects on the medial (17 patients) and central (1 patient) regions of the trochlea were observed at a mean of 3.4 months after the trauma. They also persisted up to 4 to 7 years (latest follow-up in an intermediate follow-up group) and eventually reossified by skeletal maturity (in a long-term follow-up group). Magnetic resonance imaging was performed on eight patients after a mean of 1.9 years from injury. Low signal intensity on T2 indicative of cartilage necrosis and an intact articular surface were found. Cubitus varus deformity, which developed in almost all patients, was observed to be nonprogressive after 3 to 4 years but persisted until maturity. Limitation of motion and late neuropathy were not seen. PMID- 11856929 TI - Displaced fractures of the lateral humeral condyle: criteria for implant removal. AB - Displaced fractures of the lateral humeral condyle have been successfully treated with closed or open reduction and pinning. However, there is no consensus as to when the implants should be removed, with the recommended time ranging from 3 to 8 weeks. The purpose of this study was to assess the results after treatment of displaced lateral condyle fractures of the humerus in children and formulate guidelines for removing the implants. Fifty-five patients with displaced fractures were included in this study. A modified Hardacre functional rating system was used to grade the final clinical result. In 50 patients the fracture was clinically and radiographically healed at 6 weeks, in 4 patients at 5 weeks, and in only 1 patient at 4 weeks. The implants should not be removed until healing can be demonstrated radiographically. This time averaged 6 or more weeks in most of the patients in this study. PMID- 11856930 TI - Posterior instability of the shoulder after supracondylar fractures recovered with cubitus varus deformity. AB - Cubitus varus is a common complication after supracondylar fractures of the humerus, and there have been several discussions about the timing of correction of deformity. Although surgery is performed mainly for cosmetic purposes, the authors in this article show the relationship between cubitus varus and dislocation of the ulnar nerve and posterior instability of the ipsilateral shoulder with a Bankart lesion in three children. A special type of osteotomy to obtain three-dimensional correction was made. All shoulders were found to be clinically stable at follow-up with full pain-free range of motion. Corrective osteotomy of the distal humerus itself corrected the varus angulation and dislocation of the medial portion of the triceps in two patients and prevented the ulnar nerve from dislocating. The authors' experience illustrates the importance of biomechanics in understanding the pathoanatomy of cubitus varus; this deformity should not be regarded as a cosmetic deformity and should be treated early. PMID- 11856931 TI - Delayed treatment of type 3 supracondylar humerus fractures in children. AB - A retrospective review of 158 type 3 supracondylar humerus fractures was undertaken to determine whether any correlation exists between an increased time from injury to surgery and four unfavorable results: a longer operative time, an increase in hospital stay, an increase in the need to open the fracture, or an increase in unsatisfactory outcomes. The average age of the patients was 5.0 years. Five children had nerve injury on initial examination, and no arm was poorly perfused. The average time from injury to evaluation in the emergency department was 9.8 hours and the average time from the emergency department to surgery was 11.5 hours. The average total time from injury to surgical treatment was 21.3 hours. The patients were in the hospital between 1 to 6 days. The average operative time was 53 minutes. Thirty patients had unsatisfactory results, defined as a pin infection, more than 15 degrees loss of motion in any plane, loss of normal carrying angle, neuropraxia, or retained hardware. There was no correlation between an increase in time to surgical intervention and longer operative time or need to open the fracture site, nor was there an indication that the delay to surgical treatment resulted in a longer hospital stay or an increase in unsatisfactory results. PMID- 11856932 TI - Early versus late reduction of a physeal fracture in an animal model. AB - This study was designed to determine whether delayed reduction of physeal fractures in an animal model causes growth disturbance, and whether final alignment is better in delayed or malreduced fractures. Salter 1 fractures of the proximal tibia were created in 41 immature rats randomized into five groups. The fractures were reduced as follows: group 1, immediately; group 2, 6 hours; group 3, 24 hours; group 4, 48 hours; and group 5, left malreduced. Both legs were analyzed for leg-length discrepancy, angular deformity, and evidence of radiographic bar. No radiographic physeal bar or leg-length discrepancy was seen among any of the groups. Angular deformity was greatest in group 5 and least in group 1. Immediate reduction resulted in the least angular deformity. Delayed reduction showed no evidence of physeal damage, physeal growth disturbance, or radiographic bar formation. Alignment was not improved in delayed reduction versus malreduced fractures. PMID- 11856933 TI - Magnetic resonance imaging analysis of hip joint development in patients with diastrophic dysplasia. AB - The authors used magnetic resonance imaging to evaluate the hip joint in patients with diastrophic dysplasia. T1- and T2-weighted images were obtained from 35 patients (25 female patients, 10 male patients) of different ages (1-39 years). The status of the joint and paraarticular soft tissues was evaluated. Hip joint congruity was good, although the joint was usually deformed. The thickness of the joint cartilage was diminished and signs of early osteoarthritis, including bone cysts and local edema, were common. The ligamentum teres was visible in only 24% of patients, suggesting abnormality of the ligamentous structures. Epiphyses were flattened or absent in all young patients. Of the 17 visible epiphyses, 7 showed avascular necrosis, indicated as a decrease in signal intensity in both T1- and T2-weighted images. It seems that proximal femoral epiphysis fails to bear normal weight pressure. PMID- 11856934 TI - Ossification of femoral head: normal sonographic standards. AB - The aim of the study was to establish normal reference standards for the appearance of the femoral head ossification center according to age, sex, and gestational age. Sonographic examination of the hip was performed in 1,800 healthy Indian and Israeli infants (900 each) aged 2 to 24 weeks. There were an equal number of boys and girls. The ossification center was noted at 2 weeks of life in the Israeli infants and at 8 weeks in the Indian infants. However, from 10 to 16 weeks, it was noted in 81% or more of the Indian infants but only 22% to 74% of the Israeli ones. In both groups, between 20 and 24 weeks of age, it was noted in more than 90% of the infants. An association was found between gestational age at birth and chronologic age of appearance of the femoral head ossification center, but only among the Israeli infants. There were no differences between genders. Knowledge of the normal sonographic appearance of the femoral head ossification center by age and ethnicity will help clinicians in the diagnosis of hip disorders. PMID- 11856935 TI - Posterior acetabular spike: an obstacle to reduction in long-standing hip dislocations. AB - The posterior insertion of the transverse acetabular ligament can be transformed into a toothlike bony prominence narrowing the acetabulum in its distal part. This deformity can impede the placement of the femoral head deep in the acetabulum, making a complete reduction impossible. The toothlike spike can be detected by computed tomography or by intraoperative palpation, and can be corrected by pushing it back with a punch under palpatory control. PMID- 11856936 TI - Development of the acetabulum and the acetabular labrum in the normal child: analysis with radial-sequence magnetic resonance imaging. AB - The degree of acetabular and labral coverage was clarified in children at different positions using radial-sequence magnetic resonance imaging. Scans were performed on 40 hips in 20 normal children (group A, 6-8 years old; group B, 9-11 years old; group C, 12-13 years old) and on 19 hips of 10 healthy adults as controls. The degrees of coverage of the femoral head by acetabulum, acetabular labrum, and both were measured at seven positions at every 15 degrees from anterosuperior 45 degrees via midsuperior to posterosuperior 45 degrees. Coverage of the femoral head by the acetabulum in young children was less than in adults at all positions, but the total coverage including the labrum was greater than in adults. Development of the acetabulum and the acetabular labrum showed differences by position. PMID- 11856937 TI - How should the acetabular angle of Sharp be measured on a pelvic radiograph? AB - Four experienced orthopaedic surgeons measured the acetabular angle of Sharp (AA) in 66 hips by using two methods. In the classic method (AA-c), the lateral margin of the acetabular roof was used as the landmark. In the modified method (AA-m), the lateral margin of the sourcil (subchondral bony condensation in the acetabular roof) was used as the landmark. Mean AA-c and AA-m measurements were 44.8 degrees and 48.4 degrees, respectively. Mean intraobserver variations of AA c and AA-m were 1.8 degrees and 2.1 degrees, respectively. Mean interobserver variations of AA-c and AA-m were 2.1 degrees and 2.6 degrees, respectively. Means obtained using AA-c of normal and defective acetabular roofs were similar, but the difference between AA-m means of different acetabular roof types was significant. The authors conclude that both AA-c and AA-m were reliable measurement methods, but clinical use of AA-m would probably produce better understanding of the radiographic hip anatomy in dysplastic hips. PMID- 11856938 TI - Three-dimensional assessment of the hip joint by two-directional arthrography. AB - This retrospective study involved assessment of the three-dimensional bony and cartilaginous cover of the hip acetabular shelf of the hip joint using arthrograms. The extent of lateral and anteroposterior cover, the time course of the changes of cover in each direction, and the relationship between lateral and anteroposterior cover were investigated. It was found that the final extent of acetabular cover could be predicted sooner from the cartilaginous cover than from the bony cover on both the contralateral side and the affected side. There was no significant correlation between lateral and anteroposterior cover on the contralateral side, suggesting the need for assessment of anteroposterior cover to avoid a late diagnosis of anterior acetabular dysplasia. PMID- 11856939 TI - Progressive slippage after pinning for slipped capital femoral epiphysis. AB - The authors retrospectively reviewed seven cases of progressive slipped capital femoral epiphysis after screw fixation. All seven patients initially presented with chronic symptoms, and five had an acute exacerbation of symptoms with the appearance of an acute-on-chronic slip. Of the other two, one had obvious motion at the proximal femoral physis and the other had increased symptoms but did not have an obvious acute slip radiographically. All underwent percutaneous screw fixation. In four patients a single screw was placed, and in three patients two screws were placed. No patient became symptom-free after surgery. Slip progression was noted on average 5 months after treatment. Radiographs in all patients revealed an increase in slip severity and loss of screw purchase in the femoral neck while fixation in the proximal femoral epiphysis remained secure. One patient had hypothyroidism and another Cushing disease, both diagnosed after the slipped epiphysis. Slips occurring in children with underlying endocrinopathies, and unstable slips in children with a history of antecedent knee or hip pain (commonly called an acute-on-chronic slip) may be susceptible to screw fixation failure. In such patients, close radiographic follow-up, particularly in the presence of continued symptoms, is required to document slip progression and fixation failure as soon as possible. PMID- 11856941 TI - Risk of refracture through unicameral bone cysts of the proximal femur. AB - The authors determined the results after traction or traction and hip spica treatment of the initial fractures through unicameral bone cysts of the proximal femur in 20 children. All of the eight displaced fractures healed, but with coxa vara and avascular necrosis in one, coxa vara in a second, and coxa breva in a third. Spontaneous healing of the cyst occurred in three of the eight children; satisfactory healing was achieved and maintained after intralesional corticosteroid injections in four of the eight children. In the remaining child with a displaced fracture, reactivation of the cyst and exercise-related pain, indicative of an incipient refracture, occurred 3 years after initial presentation. All of the 12 undisplaced fractures healed without deformity or avascular necrosis. Intralesional corticosteroids were used in all of the 12 children because none of them showed spontaneous healing of their cysts. Satisfactory radiographic healing was achieved 1 year after presentation in all of the 12 children. However, one or more refractures resulting from reactivation of the cyst occurred in 6 of the 12 children 2 to 5 years after initial presentation. The results of this study indicate that satisfactory radiographic healing needs to be achieved by the end of the first year and needs to be maintained thereafter to prevent refractures. PMID- 11856940 TI - Scope and limitations of limb-sparing surgery in childhood sarcomas. AB - The authors reviewed 43 children with high-grade sarcomas of bone and soft tissue who underwent surgery during the past 9 years. Twenty-six patients had osteosarcoma, 7 had Ewing sarcoma, and 10 had soft tissue sarcomas. Patients ranged in age from 4 months to 13 years. Mean follow-up period was 6 years. The feasibility of resection, results of local resection, and failure of local disease control were reviewed. Limb salvage was feasible in 81% of patients. Of the 35 patients undergoing limb salvage, margins were considered adequate in 88% and inadequate in 12%. Limb-sparing resection was not feasible in 19% of patients. Five patients (four with metastatic disease at presentation) did not undergo surgery and were given palliative care; three other patients underwent amputation. The decision of whether to proceed with limb salvage surgery must depend on the aggressiveness of the underlying tumor, its stage, the feasibility of obtaining tumor-free resection margins, and the response to neoadjuvant therapy. PMID- 11856942 TI - Intramedullary rodding and bisphosphonate treatment of polyostotic fibrous dysplasia associated with the McCune-Albright syndrome. AB - The authors report the results of the management of five consecutive children with McCune-Albright syndrome. These children were treated with a combination of drug treatment (bisphosphonates) and surgical treatment with elongating intramedullary rods (Sheffield) for management of femoral and tibial lesions. This treatment was successful in all patients, as judged by improvement in their quality of life and in clinical parameters, such as decreased bone pain and fracture rate, and improved walking ability. Two of the five children had been wheelchair-bound before treatment. All children are now community ambulators. In 5 of 10 hips, there was a significant decrease in the neck-shaft angle over time compared with the immediate postoperative angle. PMID- 11856943 TI - Use of the Internet by pediatric orthopaedic outpatients. AB - The Internet is a rich source of medical information, but little is known about orthopaedic patients' use of the Internet for medical information. Adults accompanying children to the clinic of a pediatric orthopaedist were surveyed regarding their rates of access to and use of the Internet for orthopaedic information and their opinions about the success of their searches and the utility of the Internet as a source of medical information. The response rate was 84% (212/253). Search rates were lowest for patients with fractures (18.2%) and highest for patients with scoliosis (53.5%). Slightly more than three quarters found their search helpful, and slightly less than half planned to discuss their search with the physician. Use of the Internet by patients and family members can create additional time demands on the orthopaedist with questions resulting from their search. Physicians can minimize time demands and educate their patients by guiding them to reputable, patient-oriented Internet sites and taking part in the creation of such sites. PMID- 11856944 TI - Quality of evaluation and management of children requiring timely orthopaedic surgery before admission to a tertiary pediatric facility. AB - The purpose of this study was to investigate the quality of evaluation and management of children requiring timely orthopaedic surgery before admission to a tertiary pediatric facility. A retrospective chart review was performed on 372 consecutive children who underwent orthopaedic surgery for a diagnosis of fracture, infection, slipped capital femoral epiphysis (SCFE), or compartment syndrome during a 22-month period at the authors' facility. Of the 372 children, 142 children (38%) first received medical care elsewhere; these are the subjects of this study. Before being seen at the authors' institution, 27 children (19%) had a problem in treatment or diagnosis and 18 (13%) had a delay in diagnosis or treatment of greater than 48 hours. Problems in diagnosis included five children with unrecognized open fractures, four of whom did not receive antibiotics; six children with missed SCFE, five of whom were not made non-weight-bearing; and six missed closed fractures. Delay in treatment occurred for 15 fractures, with a mean delay of >7 days until surgical treatment. Insurance status and primary language of the family were not associated with problems or delays in treatment. Overall, 32% of children undergoing time-sensitive orthopaedic surgery at a tertiary pediatric center had problems or delays in the medical care they received before transfer. PMID- 11856945 TI - Common benign lesions of bone in children and adolescents. AB - Although benign bone neoplasms are relatively uncommon in children and adolescents, their discovery can produce great angst among providers and parents. In most but not all instances, imaging studies can indicate the likely benign nature of the tumor and allay fears. The clinical presentation, imaging, and contemporary management of five common types of tumor or tumor-like conditions of bone commonly seen in children and adolescents are reviewed: nonossifying fibroma, unicameral bone cyst, aneurysmal bone cyst, fibrous dysplasia, and exostosis. PMID- 11856946 TI - Effect of lower limb Sofield procedure on ambulation of osteogenesis imperfecta. PMID- 11856948 TI - Prevention and treatment of injury from chemical warfare agents. PMID- 11856949 TI - Cefditoren (Spectracef)--a new cephalosporin. PMID- 11856950 TI - Budesonide (Entocort EC) for Crohn's disease. PMID- 11856951 TI - Ortho Evra--a contraceptive patch. PMID- 11856952 TI - Activated protein C (Xigris) for severe sepsis. PMID- 11856953 TI - Anakinra (Kineret) for rheumatoid arthritis. PMID- 11856954 TI - Almotriptan (Axert) and frovatriptan (Frova) for migrane. PMID- 11856955 TI - Effects of running exercise with increasing loads on tibialis anterior muscle fibres in mice. AB - Cross-sectional areas and succinate dehydrogenase (SDH) activities of type identified fibres in the deep, middle and superficial regions of the tibialis anterior muscle in mice were examined after 4 weeks of voluntary running exercise with increasing loads. Nineteen-week-old male mice were assigned randomly to either a control or exercise group. The mean cross-sectional areas of all types (IIa, IIx and IIb) of fibres in the superficial region of the muscle were greater in the exercise group than in the control group. The mean SDH activities of type IIx and type IIb fibres in the middle region and of all types (IIa, IIx and IIb) of fibres in the superficial region of the muscle were greater in the exercise group than in the control group. These results suggest that voluntary running exercise with increasing loads causes hypertrophy and/or an increase in the SDH activity of fibres in the specific muscle region where fibres with a high threshold and a low-oxidative enzyme activity are distributed, and these fibres are recruited to adapt to changes in exercise conditions. PMID- 11856956 TI - Noxious somatic inputs to hypothalamic-midbrain projection neurones: a comparison of the columnar organisation of somatic and visceral inputs to the periaqueductal grey in the rat. AB - The induction of Fos protein was used to localise hypothalamic neurones activated by noxious somatic stimulation. This was combined with retrograde transport of fluorescent latex microspheres from identified 'pressor' and 'depressor' sites in the dorsolateral/lateral or ventrolateral columns of the periaqueductal grey (PAG). Fos-positive neurones were found throughout the rostral hypothalamus. Of those neurones activated by noxious somatic stimuli that projected to the PAG all but one was retrogradely labelled from sites that included the lateral column. Only one neurone was double labelled following injection of tracer at a depressor site in the ventrolateral PAG. This is in marked contrast to visceroresponsive hypothalamic neurones, a larger proportion of which project to the PAG and which, as reported previously, preferentially target depressor sites in the ventrolateral sector. These results are discussed in relation to the roles of the anterior hypothalamus and the different functional columns of the PAG in co ordinating autonomic and sensory functions in response to nociceptive inputs originating in different peripheral domains. PMID- 11856957 TI - C-nociceptor activation of hypothalamic neurones and the columnar organisation of their projections to the periaqueductal grey in the rat. AB - The induction of Fos protein was used to localise hypothalamic neurones activated by ramps of noxious skin heating delivered at a rate of 2.5 degrees C s(-1) to preferentially activate C-nociceptors. This was combined with retrograde transport of cholera toxin subunit B from identified 'pressor' and 'depressor' sites in the dorsolateral/lateral or the ventrolateral columns of the periaqueductal grey. Fos-positive neurones were found throughout the rostral hypothalamus. Despite this wide distribution, those neurones double labelled retrogradely from the periaqueductal grey were focused in the lateral area of the anterior hypothalamus. More than 20 % of Fos-positive neurones in this region projected to depressor sites in the ventrolateral periaqueductal grey, and 10 % projected to its dorsolateral/lateral sector. These results are discussed in relation to the peripheral inputs to hypothalamic-midbrain pathways and their role in the cardiovascular responses to different components of the pain signal. PMID- 11856958 TI - Pyeloureteral motility and ureteral peristalsis: essential role of sensory nerves and endogenous prostaglandins. AB - The cellular mechanisms that underlie the initiation and propagation of the peristaltic contractions, which transport urine from the kidney to the bladder for storage, remain little understood. Extracellular and intracellular microelectrode recordings have identified two populations of smooth muscle cells as well as a population of renal interstitial cells (RICs) that all display spontaneous electrical activity. By analogy with the heart it has been proposed that atypical smooth muscle cells, preferentially located in the very proximal regions of the renal pelvis, generate the essential pacemaker signal. These pacemaker potentials propagate to neighbouring typical smooth muscle cells or RICs to trigger action potential discharge. These action potentials then propagate distally to trigger other bundles of typical smooth muscle cells. The frequency of action potential discharge and contraction decreases as the relative number of RICs and atypical smooth muscle cells compared to typical smooth muscle cells decreases with distance from the renal fornix. It is clear that functional capsaicin-sensitive sensory afferents and the endogenous release of both tachykinins and prostaglandins are essential in the maintenance of normal peristalsis, as well as in monitoring and responding to any chemical or mechanical stimulation. However, the cellular mechanisms underlying the action of these endogenously-released agents remain to be elucidated. PMID- 11856959 TI - Muscarinic agonist-induced non-granular and granular secretion of amylase in the parotid gland of the anaesthetized rat. AB - The muscarinic agonist bethanechol was infused intravenously, under alpha- and beta-adrenoceptor blockade, in anaesthetized rats at various dose levels (5-10, 20 and 40-50 microg kg(-1) min(-1)) over 30 min. The amount of saliva secreted from the parotid gland was dose dependent at 95, 202 and 737microl, respectively. The salivary amylase activity was approximately the same at the two lower doses (506 U and 448 U, respectively), while it was higher (1268 U) at the highest dose. In response to the highest dose, but not to the lower doses, the total parotid glandular amylase activity and the numerical density of parotid acinar secretory granules were lowered, by 25 % and 22 %, respectively. Thus, in the rat parotid gland, agonists such as bethanechol, which use Ca(2+) as a second messenger, may release proteins not only by non-granular mechanisms but also, and in contrast to the general belief, by granule exocytosis. PMID- 11856960 TI - Regional expression of inducible nitric oxide synthase in the kidney stimulated by lipopolysaccharide in the rat. AB - The renal medulla contains more mRNA of the inducible isoform of nitric oxide synthase (iNOS) than the cortex, which may be important in preventing ischaemic injury, since blood flow and tissue oxygen tension are normally low in this region. We examined the effects of the bacterial endotoxin E. coli lipopolysaccharide (LPS) on renal function and regional expression of iNOS in male Sprague-Dawley rats. In six rats, glomerular filtration rate (GFR) and effective renal plasma flow (ERPF) were 0.95 +/- 0.09 ml min(-1) g(-1) and 3.36 +/- 0.20 ml min(-1) g(-1), respectively, and decreased significantly to 0.35 +/- 0.09 and 1.74 +/- 0.54 ml min(-1) g(-1), respectively, 1 h after administration of LPS. In an additional seven rats, GFR and ERPF were 0.91 +/- 0.07 and 2.97 +/- 0.30 ml min(-1) g(-1), respectively, 18 h after LPS administration; these values were similar to those in control rats. In all rats, arterial pressure was stable throughout all study periods. In control rats, immunoblot analysis revealed expression of the iNOS protein in the cortex and more pronounced expression in the medulla. In rats studied 18 h after LPS treatment, there was a striking increase in the iNOS expression in the outer medulla. Immunohistochemical examination in the LPS-treated rats showed limited iNOS immunostaining in the cortex, localised to the vascular endothelium and macula densa; however, intense and widespread staining was noted in the tubular and vascular structures of the outer medulla. These findings demonstrated a differential constitutive expression of iNOS protein in different regions of the rat kidney, and marked augmentation of iNOS expression in the outer medulla by LPS. PMID- 11856961 TI - A ratiometric method of autofluorescence correction used for the quantification of Evans blue dye fluorescence in rabbit arterial tissues. AB - Evans blue dye (EBD) conjugates with albumin in the circulation and is frequently used to measure vascular protein leakage. The fluorescence of the dye from tissue sections can be used to measure its uptake at very specific anatomical locations, but problems arise with dye quantification because tissue components also fluoresce; so-called autofluorescence. We have measured uptake of EBD by blood vessel walls at various points around the aorto-renal branch of rabbits. High resolution, digitised, fluorescence images of histological sections of artery wall allowed detailed microscopic analysis of EBD accumulation; and a ratiometric method was developed to enable autofluorescence to be separated from EBD fluorescence. When EBD-free tissue sections were illuminated with blue light, the ratio of red to green fluorescence was constant throughout the tissue (0.59 +/- 0.03, mean +/- S.D., n = 32). Therefore, at each individual pixel, the level of red autofluorescence could be determined by multiplying the green intensity at that pixel by the calculated red to green ratio. Since EBD fluorescence was detected only in the red region of the spectrum, intensity values of the dye alone were obtained from EBD-exposed tissue by subtracting the red autofluorescence estimated by this ratiometric method. In such cases the red to green fluorescence ratio was measured from adjacent sites known to be free of EDB (0.59 +/- 0.02, mean +/- S.D., n = 56). We were therefore able to increase the sensitivity of tracer quantification by complete elimination of background autofluorescence on a pixel-by-pixel basis. Use of EBD standards allowed calibration of corrected fluorescence intensities and calculation of mass transfer coefficients for albumin into the artery wall. Spatial variations in the permeability of the artery wall around the renal ostium were detected with the present high resolution technique, with an average mass transfer coefficient of (6.8 +/- 0.9) x 10(-8) cm s(-1) for all sites combined (n = 56). The present ratiometric method could potentially be applied to other quantitative fluorescence-based techniques. PMID- 11856962 TI - Analysis of the effects of graded levels of hypoxia on noradrenaline-evoked contraction in the rat iliac artery in vitro. AB - In rings of rat iliac artery, contractions were evoked by noradrenaline (NA), the selective alpha(1) adrenoceptor agonist phenylephrine (PE), and K(+), which causes depolarisation-induced contraction. There was no evidence of alpha(2) adrenoceptor-evoked contraction. Hypoxia, induced by reducing P(O(2)) in the bath from 100 mmHg to 70, 55 or 40 mmHg, had similar effects on rings with (E+) and without (E-) endothelium. In E- rings, the NA concentration-response curve was biphasic, whereas that for PE was monophasic. Hypoxia reduced maximum contractions in response to NA and PE (NA(max) and PE(max), respectively) without affecting the concentrations that evoked 50 % of maximum contraction (EC(50)). At P(O(2)) of 70 mmHg, NA(max) of the high affinity alpha(1) receptor for NA (NA(maxh)) and PE(max) were reduced by approximately 15 %, but at P(O(2)) of 55 and 40 mmHg, NA(maxh) was severely attenuated while PE(max) fell by 45 and 75 %, respectively. Similarly, the Ca(2+) channel blocker nicardipine depressed NA(maxh) and PE(max), but P(O(2)) of 55 mmHg further reduced NA(max) and PE(max). Hypoxia also reduced contractions evoked by NA, PE or K(+) at the concentrations required to produce 80 % of the maximum contraction (EC(80)), receptor-mediated contractions being more affected. Ca(2+)-free conditions reduced the contractions evoked by NA and PE, at the EC(80), to approximately 10 % of control. The K(+) channel inhibitors glibenclamide and tetraethylammonium did not prevent hypoxia induced depression of PE-evoked contraction. Thus, contractions evoked in iliac artery by the high affinity subtype of alpha(1) adrenoceptor for NA, which may respond to circulating levels of NA, and by the single alpha(1) adrenoceptor subtype for PE, are especially vulnerable to P(O(2)) levels less-than-or-equal 55 mmHg. We propose that this reflects hypoxia-induced inhibition of Ca(2+) influx through L-type and receptor-operated Ca(2+) channels; K(+) channel opening makes little contribution. PMID- 11856963 TI - Mechanical control of extracellular space in rabbit atria: an intimate modulator of the translocation of extracellular fluid and released atrial natriuretic peptide. AB - We have previously shown that extracellular fluid (ECF) is translocated by atrial contraction. Following on from this finding we have proposed a two-step sequential mechanism for the regulation of stretch-activated secretion of atrial natriuretic peptide (ANP): myocytic release of ANP into the surrounding paracellular space followed by the translocation of ECF with the released ANP into the bloodstream. This latter step is induced by atrial contraction, and is therefore controlled by atrial workload. However, the mechanism that regulates the changes in translocation of the ECF has not been defined. To define the relationship between the atrial workload, ECF translocation, size of the extracellular space (ECS) and ANP secretion, experiments have been performed in isolated perfused beating rabbit atria. Atrial workload, transendocardial translocation of the ECF and the secretion of ANP were quantified. Changes in the size of the atrium and the ECS were determined by a newly developed methodology in the beating atria. Atrial workload determined the translocation of the ECF and released ANP with waning of the translocation at higher myocardial workloads. Atrial workload inversely determined the size of the atrium and the ECS. The latter directly determined the translocation of the ECF in terms of atrial workload. From these data we suggest that the size of the ECS is an intimate modulator of the translocation of the ECF and released ANP, and that the phenomenon of waning of the transendocardial translocation that appeared at higher atrial workloads is closely related to the shrinkage of the ECS. PMID- 11856964 TI - Hypertension-related intermyocyte junction remodelling is associated with a higher incidence of low-K(+)-induced lethal arrhythmias in isolated rat heart. AB - The aim of this study was to characterise the arrhythmogenic mechanisms involved in hypokalaemia-induced sustained ventricular fibrillation (SVF), in hypertensive rats. The hearts from rats with hypertension induced by the nitric oxide synthase inhibitor L-NAME, and age-matched normotensive controls, were perfused in Langendorff mode with oxygenated Krebs-Henseleit solution followed by a K(+) deficient solution. In additional experiments, free intracellular Ca(2+) concentration ([Ca(2+)](i)) was measured using fura-2 in conjunction with an epicardial optical probe. The epicardial electrocardiogram was continuously monitored during all experiments. The gap junction protein connexin-43 and the ultrastructure of the cardiomyocytes were examined, and selected enzyme activities were measured in situ. There was a higher incidence of low-K(+) induced SVF in the hearts of hypertensive compared to normotensive rats (83 % vs. 33 %, P < 0.05). Perfusion with a low-K(+)-containing solution lead to elevation of diastolic [Ca(2+)](i) that was accompanied by premature beats, bigeminy, ventricular tachycardia and transient ventricular fibrillation. These events occurred earlier with increased incidence and duration in the hearts of hypertensive rats (arrhythmia scores: hypertensive, 4.9 +/- 0.7; normotensive, 3.1 +/- 0.1; P < 0.05), which exhibited apparent remodelling accompanied by a significant decrease in the density of connexin-43-positive gap junctions. Moreover, low-K(+)-related myocardial changes, including local impairment of intermyocyte junctions, ultrastructural alterations due to Ca(2+) overload and intercellular uncoupling, and decreased enzyme activities were more pronounced and more dispersed in hypertensive than normotensive rats. In conclusion, nitric oxide-deficient hypertension is associated with decreased myocardial coupling at gap junctions. The further localised deterioration of junctional coupling, due to low-K(+)-induced Ca(2+) disturbances, as well as spatial heterogeneity of myocardial alterations including interstitial fibrosis, probably provide the mechanisms for re-entry and sustaining ventricular fibrillation. PMID- 11856965 TI - Milk prolactin, feed volume and duration between feeds in women breastfeeding their full-term infants over a 24 h period. AB - Previous studies have suggested that the uptake of prolactin from the blood into the milk may be restricted when the alveolus is distended with milk. Therefore the aim of this study was to determine the relationship between prolactin in milk and milk production by measuring the concentration of prolactin in samples of fore- and hind-milk as well as the volume of milk removed for each breast, at each breastfeed over a 24 h period. The mean (+/- S.D.) concentration of prolactin in milk for all women (n = 15) over the 24 h period was 18.5 +/- 11.6 microg l(-1) (fore-milk) and 16.8 +/- 12.8 microg l(-1) (hind-milk). The variation between women masked small changes within women in the concentration of prolactin in milk over the 24 h period, therefore a prolactin ratio (individual fore- or hind-milk value divided by the mean for all fore- or hind-milk samples collected over a 24 h period) was determined. The concentration of prolactin was highest in milk between 02.01 and 06.00 h (prolactin ratio for fore- to hind milk, 1.5), and lowest between 10.01 and 18.00 h (prolactin ratio for fore- to hind-milk, 0.8). Furthermore, we observed that the difference in prolactin concentration between the fore- and hind-milk (fore-hind gradient) was greatest between 06.01 and 10.00 h (4 microg l(-1)). To ensure that this effect was not due to permeability in the paracellular pathway, the concentrations of serum albumin and sodium in milk were measured. No significant (P > 0.05) changes over the 24 h period, or with increasing time since last feed were observed. We therefore concluded that when the breast is most drained of milk (in the late evening), and the rate of milk synthesis is greatest, the fore-hind prolactin gradient in the milk of the following feed (in the early morning) is highest. This is consistent with the observation that prolactin uptake from the blood by the lactocyte is dependent on the fullness of the breast, such that prolactin uptake may be inhibited in full alveoli. PMID- 11856966 TI - Hyperprolactinaemia during prolonged exercise in the heat: evidence for a centrally mediated component of fatigue in trained cyclists. AB - Several lines of evidence suggest that central serotoninergic neurons may mediate fatigue signals during prolonged exercise. In this study we examined the effects of diet and ambient temperature on peripheral modulators and indices of serotoninergic function and their relationship to exercise performance. Six well trained cyclists participated, in randomised order, in two diet and exercise regimens each lasting 8 days and comprising four cycle tests to exhaustion at 70 % of maximum oxygen uptake. On days 1 and 5, subjects exercised to exhaustion to deplete muscle glycogen. For 3 days after the first depletion trial a diet providing 10 % of energy in the form of carbohydrate (CHO) was consumed (low CHO), and for 3 days after the second depletion trial a diet providing 80 % (high CHO) of energy as CHO was consumed, and each diet was followed by a performance trial at the same ambient temperature, either 10 degrees C or 30 degrees C (days 4 and 8). This schedule was repeated after 1 week, but performance trials were carried out at the other ambient temperature. In the cold, cycling time increased (P < 0.01) from 89.2 (78.0-129.5) min (median (range)) in the low CHO trial to 158.2 (116.9-165.6) min in the high CHO trial. In the heat, cycling time increased from 44.0 (31.8-51.4) min in the low CHO trial to 53.2 (50.2-82.2) min on the high CHO trial (P = 0.02). The serum prolactin (Prl) concentration was higher at exhaustion during the two trials in the heat than in the two trials in the cold. Serum Prl levels were unrelated to the purported peripheral modulators of serotoninergic function (plasma concentrations of total tryptophan (Trp), free Trp, branched-chain amino acids (BCAAs), free Trp/BCAA ratio and total Trp/BCAA ratio) but were significantly related to the rectal temperatures measured during the two trials in the heat. This finding provides indirect evidence that the serotoninergic system may be involved in fatigue during exercise under conditions of heat stress. PMID- 11856967 TI - Effects of ischaemia on subsequent exercise-induced oxygen uptake kinetics in healthy adult humans. AB - Leg muscles were occluded (33 kPa) prior to exercise to determine whether the induced metabolic changes, and reactive hyperaemia upon occlusion release just prior to the exercise, would accelerate the subsequent oxygen consumption (VO2) response. Eight subjects performed double bouts (6 min duration, 6 min rest in between) of square wave leg cycle ergometry both below and above their lactate threshold (LT). Prior to exercise, large blood pressure cuffs were put around the upper thighs. Occlusion durations were 0 min (control), 5 min and 10 min. Ischaemia was terminated within 5 s prior to exercise onset. Heart rate, VO2, ventilatory rate (V(E)), electromyogram (EMG) and haemoglobin/myoglobin (Hb/Mb) saturation were recorded continuously. Single exponential modelling demonstrated that, compared to control (time constant = 53.9 +/- 13.9 s), ischaemia quickened the VO2 response (P < 0.05) for the first bout of exercise above LT (time constant = 48.3 +/- 14.5 s) but not to any other exercise bout below or above LT. The 3-6 min integrated EMG (iEMG) slope was correlated to the 3-6 min VO2 slope (r = 0.73). Hb/Mb saturation verified the ischaemia but did not show a consistent relation to the VO2 time course. Reactive hyperaemia induced a faster VO2 response for work rates above LT. The effect, while significant, was not large considering the expected favourable metabolic and circulatory changes induced by ischaemia. PMID- 11856969 TI - Phenotype and function of somatic primary afferent nociceptive neurones with C-, Adelta- or Aalpha/beta-fibres. AB - Nociceptive dorsal root ganglion (DRG) neurones have fibres that conduct in the C, Adelta and Aalpha/beta conduction velocity range. The properties of nociceptive compared with non-nociceptive somatic afferent dorsal root ganglion neurones appear to fall into two patterns, A and B. Pattern A properties of nociceptive neurones, the more common type, include longer action potential duration and slower maximum rate of fibre firing, as well as a greater expression of substance P and calcitonin gene-related peptide immunoreactivity. The values of pattern A properties appear to be graded according to the conduction velocity group (C, Adelta or Aalpha/beta) of the fibres. The most pronounced forms of A type properties are expressed by nociceptive neurones with C-fibres, and these become less pronounced in nociceptive neurones with Adelta-fibres and least pronounced in those with Aalpha/beta fibres (C > Adelta > Aalpha/beta). Some of these properties are also expressed in a less extreme but similarly graded manner through C, Adelta and Aalpha/beta groups of non-nociceptive low threshold mechanoreceptive (LTM) neurone. The less common pattern B properties of nociceptive neurones have similar values in C-, Adelta- and Aalpha/beta-fibre nociceptive neurones but these clearly differ from LTM units with C-, Adelta- and Aalpha/beta-fibre conduction velocities. These features of nociceptive neurones include consistently larger action potential overshoots and longer after hyperpolarisation durations in nociceptive than in LTM neurones. PMID- 11856970 TI - Anatomy of primary afferents and projection neurones in the rat spinal dorsal horn with particular emphasis on substance P and the neurokinin 1 receptor. AB - The dorsal horn of the spinal cord plays an important role in transmitting information from nociceptive primary afferent neurones to the brain; however, our knowledge of its neuronal and synaptic organisation is still limited. Nociceptive afferents terminate mainly in laminae I and II and some of these contain substance P. Many projection neurones are located in lamina I and these send axons to various parts of the brain, including the caudal ventrolateral medulla (CVLM), parabrachial area, periaqueductal grey matter and thalamus. The neurokinin 1 (NK1) receptor on which substance P acts is expressed by certain neurones in the dorsal horn, including approximately 80 % of lamina I projection neurones. There is also a population of large NK1 receptor-immunoreactive neurones with cell bodies in laminae III and IV which project to the CVLM and parabrachial area. It has been shown that the lamina III/IV NK1 receptor immunoreactive projection neurones are densely and selectively innervated by substance P-containing primary afferent neurones, and there is evidence that these afferents also target lamina I projection neurones with the receptor. Both types of neurone are innervated by descending serotoninergic axons from the medullary raphe nuclei. The lamina III/IV neurones also receive numerous synapses from axons of local inhibitory interneurones which contain GABA and neuropeptide Y, and again this input shows some specificity since post-synaptic dorsal column neurones which also have cell bodies in laminae III and IV receive few contacts from neuropeptide Y-containing axons. These observations indicate that there are specific patterns of synaptic connectivity within the spinal dorsal horn. PMID- 11856971 TI - Pain pathways and parabrachial circuits in the rat. AB - This review presents a schematic attempt to classify the major pain pathways, based on the results of recent studies in our laboratory, with a special emphasis on the parabrachial system. Our view is based on results from experiments in the rat, using very small iontophoretic injections of anterograde tracers. As illustrated in this report, we have found a very dramatic difference between ascending projections originating from deep laminae compared with those arising from lamina I of the dorsal horn. We propose three main pain systems and discuss their functional-anatomical relationships. The first system is centred on the projection from deep laminae to three caudal reticular areas - the lateral reticular nucleus (LRN), the subnucleus reticularis dorsalis (SRD) and the gigantocellular lateral paragigantocellular reticular nuclei (NGc) - and the parabrachial internal lateral subnucleus (PBil). The second system is centred on the projection from lamina I to the ventral posterolateral nucleus (VPL), the ventral posteromedial (VPM), the posterior nuclear group (Po) and triangular posterior nucleus (PoT) of the thalamus. The third system is centred on the projection from lamina I to the lateral parabrachial area. We also present the four main projections from the latter area to the extended amygdala, the hypothalamus, the periaqueductal grey matter (PAG), and the ventrolateral medulla (VLM), and their involvement in emotional and autonomic (homeostatic) aspects of pain. PMID- 11856972 TI - The nucleus of the solitary tract: an integrating station for nociceptive and cardiorespiratory afferents. AB - Painful stimuli can evoke dramatic responses in the cardiovascular and respiratory systems. We have assessed the role of both the sympathetic and parasympathetic nervous system in mediating the reflex tachycardia that accompanies somatic nociception. We describe a major role for the nucleus tractus solitarii (NTS) as a site for integrating nociceptive and cardiorespiratory afferents. Since cardiorespiratory and nociceptive afferents terminate in the NTS, this nucleus offers a powerful opportunity for central modulation. We show that the NTS plays a major role in mediating the reflex tachycardia evoked by somatic noxious stimulation. Similar noxious stimulation attenuates the cardiac component of the peripheral chemoreceptor reflex and inhibits the peripheral chemoreceptor-evoked excitatory synaptic response of some NTS neurones. The functional interpretation we propose is that by depressing homeostatic reflexes at the NTS, noxious stimulation-evoked cardiorespiratory changes can be expressed and maintained, which may be essential for the survival of the animal. PMID- 11856973 TI - The caudal medullary ventrolateral reticular formation in nociceptive cardiovascular integration. An experimental study in the rat. AB - The endogenous pain control system is composed of multiple functionally distinct brain regions, which are thought to integrate nociceptive information with various brain functions. The clear involvement of some pain control centres in cardiovascular modulation has been claimed as a strong indication of their role in nociceptive-cardiovascular integration. Particular emphasis has been given to their putative function in triggering cardiovascular reactions to pain. However, the possibility of their participation in the less-studied influence of cardiovascular conditions in pain perception has been largely ignored. We have recently addressed this issue by investigating the involvement of the caudal ventrolateral medullary reticular formation (cVLM) in hypertension-induced hypoalgesia. Circuits capable of conveying cVLM-elicited antinociception include a direct reciprocal cVLM-spinal loop, and two disynaptic spinal pathways relaying in rostroventromedial medullary (RVM) neurones and A(5) noradrenergic neurones. In the three pathways, the cVLM neurones involved are circumscribed to a small area of reticular formation located laterally to the lateral reticular nucleus, the VLMlat. The VLMlat has a vasodepressor effect similar to that obtained from the cVLM. In the spinal cord dorsal horn, c-fos expression evoked by noxious stimuli is decreased in hypertensive animals, as compared to normotensive animals. In hypertensive animals following lesion of the VLMlat, spinal c-fos expression is identical to that observed in normotensive animals. The collected data point to a role for the VLMlat in the depression of spinal nociceptive processing in response to rises in blood pressure. Since hypertension-induced hypoalgesia is mediated by spinal alpha(2)-adrenoreceptors, this effect could be conveyed by the cVLM-A(5)-spinal pathway. PMID- 11856974 TI - Distinct central representations of inescapable and escapable pain: observations and speculation. AB - It is well established clinically that the affective response to pain of deep origin (muscles, joints and viscera) is distinct from that evoked by cutaneous pain. Cutaneous pain triggers a fight-flight reaction (active emotional coping), whereas deep pain evokes a reaction of quiescence, decreased vigilance and vasodepression (passive emotional coping). These observations led to suggestions of distinct central representations for deep versus cutaneous pain. Indeed, studies using immediate early gene (c-fos) expression revealed selective activation of ventrolateral versus lateral columns of the midbrain periaqueductal grey region (PAG) by persistent pain of deep origin versus intermittent cutaneous pain. Ventrolateral versus lateral PAG activation had been found earlier to evoke passive versus active emotional coping. However, not all cutaneous pain triggers active coping. Persistent cutaneous pain (e.g. burns) instead, usually evokes passive coping. This raised the question of whether the behavioural significance of pain (i.e. its escapability versus inescapability), rather than its tissue origin, is represented in supraspinal regions such as the PAG. Subsequent study revealed that a persistent (inescapable) noxious cutaneous manipulation (clip of the neck) evoked both selective ventrolateral PAG Fos expression and passive emotional coping. Such data suggest that pain representation in the PAG reflects a quality akin to behavioural significance, rather than tissue origin. In contrast, in the spinal cord predominantly superficial dorsal horn Fos expression was seen after either persistent or intermittent noxious cutaneous stimuli, leaving the question of the pathway(s) via which persistent (inescapable) cutaneous pain activates the vlPAG unanswered. One experimental approach to this question is suggested. PMID- 11856975 TI - Inescapable and escapable pain is represented in distinct hypothalamic-midbrain circuits: specific roles for Adelta- and C-nociceptors. AB - The affective responses to pain arising from deep somatic and visceral tissues differ markedly from those evoked by brief cutaneous insults. Deep pain evokes passive emotional coping that includes quiescence and vasodepression. In contrast, cutaneous pain evokes an active emotional coping: the fight or flight response. There is now considerable evidence to support the notion that nociceptive inputs arising from different peripheral domains drive the different functional columns of the periaqueductal grey (PAG) that co-ordinate either active or passive coping strategies. Nociceptive inputs from deep structures drive neurones in the ventrolateral columns that co-ordinate passive emotional coping whereas brief cutaneous insults activate the dorsolateral/lateral columns that co-ordinate active coping strategies. An emerging concept, as presented in the preceding article by Keay & Bandler, is that it is the behavioural significance of the nociceptive input, rather than its organ of origin per se, that determines the characteristics of the affective response. These authors provide evidence that brief, escapable stimuli activate neurones in the dorsolateral/lateral columns of the PAG and that inescapable, persistent pain, irrespective of its organ of origin, activates the ventrolateral column. This review will present recent evidence that differential representation of escapable and inescapable pain in the PAG extends to distinct representations of 'first' and 'second' pain, as indicated by the columnar distribution of neurones activated by inputs from Adelta- and C-nociceptors. Furthermore, the functional organisation of projections from circumscribed regions of the hypothalamus to the different columns of the PAG indicates that the behavioural significance of the pain signal is represented in brain regions other than the PAG. PMID- 11856976 TI - [Antimicrobial drug use in Spain. A problem with no solution]. PMID- 11856977 TI - [Advances in vaccines and invasive disease in children]. PMID- 11856978 TI - [Diffusion and pharmacokinetics of antibiotics in the ocular globus. Therapeutic implications]. PMID- 11856979 TI - [Use of topical antifungal agents in Spain]. PMID- 11856980 TI - [Activity of different antimicrobial agents against penicillin-resistant Streptococcus pneumoniae]. AB - Since the first strains of penicillin-resistant Streptococcus pneumoniae were isolated in the 1960s, the rate of resistance has increased world- wide, though with geographic variations. Currently, the prevalence and patterns of antibiotic resistance in this microorganism vary widely from one country to another, as well as within in the same country. In our study we evaluated the in vitro susceptibility of 299 isolates of S. pneumoniae from patients with community acquired respiratory tract infections from 1998-2000 to different antimicrobial agents. The following resistance results were obtained: 32.11% to penicillin, 4.35% to amoxicillin, 3.68% to amoxicillin-clavulanic acid, 69.9% to cefaclor, 32.44% to cefpodoxime, 34.11% to cefuroxime, and 24.41% to azithromycin. For cefixime and ceftibuten there are no NCCLS breakpoint criteria. PMID- 11856981 TI - [Clinical utility of susceptibility testing of herpes simplex virus to acyclovir]. AB - In vitro susceptibility to acyclovir of 96 strains of herpes simplex virus isolated from 80 immunocompromised patients attended in our hospital was studied by the cytopathic effect reduction assay. Ninety-eight percent (61/62) of herpes simplex virus 1 strains and 91% (31/34) of herpes simplex virus 2 strains were inhibited by acyclovir concentrations lower than 3 mg/l. In 5% of the patients herpes simplex strains resistant to acyclovir (ID(50) >3 mg/l) were isolated. Ninety-eight percent of the lesions caused by herpes simplex viruses susceptible to acyclovir (ID(50) <3 mg/l) resolved independently of treatment. In two cases, the cytopathic effect reduction assay was not able to predict treatment failure and persistance of the lesions was not always associated with isolation of a resistant strain in vitro. In four cases, isolation of a strain resistant to acyclovir was not indicative of treatment failure. In conclusion, we believe there is no need to routinely test susceptibility of herpes simplex viruses to acyclovir and that susceptibility testing should be indicated only in patients in whom lesions persist and other causes have been ruled out. PMID- 11856983 TI - [In vitro activity of six fluoroquinolones and penicillin against 101 viridans group streptococci characterized by their susceptibility to erythromycin]. AB - The increase in the use of the fluoroquinolones, including for the treatment of lower respiratory tract infections, seems to be associated with a parallel increase in bacterial resistance. We studied the activity of penicillin and six fluoroquinolones against 101 viridans group streptococci isolated from blood (58 erythromycin-susceptible and 43 erythromycin-resistant). The percentage of strains not susceptible to penicillin was 35% and there were statistically significant differences in the percentages of penicillin resistance between erythromycin-susceptible and -resistant strains. The fluoroquinolones studied showed good activity against our viridans group streptococci independent of their susceptibility to erythromycin. The norfloxacin MIC(50) and MIC(90) were 8 and 16 mg/l, respectively. Ofloxacin and ciprofloxacin showed the same activity (MIC(50) 1 mg/l and MIC(90) 2 mg/l); levofloxacin was similar with MIC(50) and MIC(90) of 1 mg/l. The fluoroquinolones with enhanced activity were moxifloxacin and trovafloxacin (MIC(50) 0.12 mg/l and MIC(90) 0.25 mg/l). However, we found two strains resistant to trovafloxacin (MIC = 4 mg/l) that showed reduced susceptibility to all the fluoroquinolones tested, including moxifloxacin (MIC = 2 mg/l). Mutations in the topoisomerase genes parC and gyrA have been shown to occur in strains with reduced susceptibility; therefore, ongoing surveillance for the development of fluoroquinolone resistance in viridans streptococci is necessary. PMID- 11856982 TI - [Activity of new fluoroquinolones against clinical isolates of Acinetobacter baumannii]. AB - Acinetobacter baumannii is an opportunistic pathogen associated with numerous nosocomial infections. In recent years it has shown extraordinary ease in developing resistance to most antimicrobial agents, which is a serious problem as it makes these infections difficult to treat. We determined the in vitro activity of eight quinolones, five betalactam agents and colistin in 160 clinical isolates of A. baumannii. In general, we observed a high rate of resistance to the quinolones (90%), excluding clinafloxacin (25%), and to ampicillin-sulbactam (61.25%) and imipenem (50%). Colistin is the agent with least resistance (13.125%), although its toxicity limits its therapeutic use. Clinafloxacin may be a good option to treat A. baumannii infections, especially in cases of therapeutic failure with other antimicrobial agents. PMID- 11856984 TI - [Community-acquired bacteremia]. AB - Bacteremia continues to be one of the main causes of mortality despite the existence of numerous antimicrobial agents and an increase in means of support. A variety of factors, such as the type of microorganism, age, the underlying disease and where the bacteremia was acquired, can change the prognosis of the infection. The aim of this study was to analyze the cases of community-acquired bacteremia gathered prospectively from the Basurto hospital in Bilbao, Spain. The incidence of bacteremia was estimated using preestablished protocol (SEPSIS-DATA) from January 1994 to September 2001. Information was gathered on all the cases of bacteremia at the hospital and only those which were of non-hospital origin (2886 cases) were selected for the study. In our hospital, 67.82% of the bacteremia cases were community-acquired, a figure which remained stable throughout the 8 year study period. A total of 54.64% of the patients were male, 1603 (55.54%) of whom were over age 60 years. The most common underlying diseases were diabetes (15.75%), neoplasia (14.96%) and HIV infection (12.9%). The majority of the cases (2216, 76.8%) were admitted to the general medicine ward, 335 (11.6%) to surgery, 200 (6.9%) to pediatrics and 135 (4.67%) to ICU. The origin of the bacteremia was urinary (33%), gastrointestinal (18%) or respiratory (18.26%). The most common microorganisms were E. coli (36%), S. pneumoniae (13%), S. aureus (9.46%), S. enteritidis (2.87%), P. aeruginosa (2.71%), P. mirabilis (2.65%) and N. meningitidis (2.45%). The bacteremia was polymicrobial in 6.27% of the cases. The most used antibiotics were ceftriaxone (31%), gentamicin (7.3%) and amoxicillin clavulanic acid (6.9%). Overall mortality was 13.82%. The relative frequency of community-acquired/nosocomial infection remained stable in two-thirds of the total cases. The incidence of bacteremia decreased in the 20-40 year age group and in those with HIV infection. E. coli and S. pneumoniae increased, while S. aureus decreased. The use of imipenem and quinolones increased. E. coli resistance to quinolones increased from 3.54% in 1995 to 14.36% in 2000. Mortality decreased slightly, with no significant differences. PMID- 11856985 TI - Inhaled steroids in chronic obstructive pulmonary disease. PMID- 11856986 TI - A year of changes--a new year of opportunities. PMID- 11856988 TI - Massive hemoptysis from pulmonary artery aneurysms. AB - Aneurysms of the pulmonary arteries are rare. The author reports a case of a patient with bilateral pulmonary artery aneurysms, in whom the nature of the lesions was not known. Pulmonary artery aneurysms should be included in the differential diagnosis of lung masses. PMID- 11856987 TI - Increased levels of airway neutrophils reduce the inhibitory effects of inhaled glucocorticosteroids on allergen-induced airway eosinophils. AB - BACKGROUND: Treatment with inhaled glucocorticosteroids attenuates allergen induced airway inflammation but is less effective in people with asthma who have noneosinophilic airway inflammation. OBJECTIVE: Studies in which glucocorticosteroid treatment was used before allergen challenges were re examined to determine whether the efficacy of steroid treatment could be predicted by baseline levels of sputum inflammatory cells. PATIENTS AND METHODS: Twenty-eight nonsmoking subjects with atopic asthma controlled by beta2-agonists participated in only one of three studies, each carried out with a double-blind, placebo controlled, randomized, crossover design. Subjects were treated with glucocorticosteroids or placebo for six to eight days and then underwent allergen inhalation challenge. Spirometry was measured for 7 h after allergen challenge, and then sputum inflammatory cells were measured. Sputum inflammatory cells were also measured before and after treatment, and 24 h after allergen challenge. The per cent inhibition of the allergen-induced airway responses by glucocorticosteroids was calculated. RESULTS: Inhaled glucocorticosteroids significantly attenuated the early and late asthmatic responses, and the number of allergen-induced sputum eosinophils (P<0.05). There was a significant negative relationship between the number of sputum neutrophils at baseline, and the per cent inhibition of allergen-induced sputum eosinophils measured at 7 h (r=-0.61, P<0.001) and 24 h (r=-0.73, P<0.0001) after challenge, suggesting that glucocorticosteroids are less effective in attenuating allergen-induced airway inflammation in subjects with high levels of neutrophils. There was no correlation between the number of sputum eosinophils at baseline and the per cent inhibition of allergen-induced responses. CONCLUSIONS: Baseline airway neutrophils, not eosinophils, can be used to predict the efficacy of inhaled steroids on allergen-induced sputum eosinophils. PMID- 11856990 TI - Editorial: Inertia in Clinical Research. PMID- 11856991 TI - Effects of Misoprostol with Interleukin-1 on Proteoglycan Metabolism of Cultured Articular Chondrocytes. AB - Bovine and normal human articular chondrocytes in suspension culture were treated with misoprostol (an analog of prostaglandin E(1) [PGE(1)]), alone and in combination with interleukin-1 (IL-1) and/or diclofenac sodium, to study effects on proteoglycan metabolism. A concentrations of 50 ng ml(minus sign1) and above, misoprostol suppressed synthesis of proteoglycans but did not affect their rate of catabolism. The mild inhibitory effect of misoprostol on proteoglycan synthesis was additive to that of IL-1, especially in chondrocytes from the superficial zone of bovine articular cartilage. In IL-1-treated cultures, diclofenac NA caused a modest improvement in proteoglycan synthesis, but this beneficial effect was diminished by the simultaneous addition of misoprostol. Cartilage or chondrocyte cultures treated with IL-1, in which proteoglycan synthesis is suppressed, serve as model systems in which to study metabolic responses of chondrocytes to potential therapeutic agents, but in these experiments, no chondroprotective effects of misoprostol were observed in IL-1 activated chondrocytes. PMID- 11856992 TI - Regulation of Collagen Gene Expression by Prostaglandins and Interleukin-1beta in Cultured Chondrocytes and Fibroblasts. AB - To compare the modulatory effects of different prostaglandins on collagen gene expression in human chondrocytes, PGE(2), PGE(1), misoprostol (PGE(1) analog), and PGF(2alpha) (10, 50 and 100 ng ml(minus sign1)) were added to human chondrocytes with or without interleukin-1beta (IL-1beta) in the presence of indomethacin to inhibit endogenous prostaglandin synthesis and the effects evaluated on chondrocyte morphology, collagen synthesis, and procollagen mRNA levels. The effects of prostaglandins on the expression of collagen gene regulatory sequences were examined using transient transfection assays of reporter gene constructs in human chondrocytes and BALB/c3T3 fibroblasts, PGE(1), misoprostol, and PGF(2alpha), similar to PGE(2), inhibited type I collagen gene expression in fibroblasts and promoted type II collagen gene expression in chondrocytes. PGE(2), the major inflammatory prostaglandin produced by IL-1 activated chondrocytes and fibroblasts, and PGF(2alpha) were somewhat more potent than the anti-inflammatory prostaglandins PGE(1) and misoprostol in counteracting the IL-1-induced suppression of type II collagen gene expression by chondrocytes and stimulation of type I collagen gene expression by fibroblasts. Rather than promoting degradation of the cartilage matrix in joint diseases, prostaglandins may be somewhat protective, suppressing fibrosis, and maintaining or promoting appropriate cartilage repair. PMID- 11856993 TI - Effects of Misoprostol and Salicylate on Canine Osteoarthritis. AB - The ability of misoprostol to reverse the deleterious changes induced in cartilage by sodium salicylate was tested using osteoarthritic canine and chondrocytes. Adult mongrel dogs were subjected to anterior cruciate ligament transection and dosed with either misoprostol, salicylate, or misoprostol plus salicylate. No significant differences were noted among the three groups in either gross and histological changes or general biochemical changes. This approach was abandoned since the levels of misoprostol attained by oral dosing were much lower than those required for domonstration of misoprostol effects in vitro. Next, chondrocytes were isolated from the osteoarthritic and contralateral knee joint cartilage of dogs 12 weeks after anterior cruciate ligament transection and cultured in alginate beads. The cultures were incubated with (3)H proline and both the genetic types of collagen synthesized and the net synthesis of (3)H-hydroxyproline were determined. No drug or combination of drugs affected the genetic types of collagen synthesized. Total protein and collagen synthesis by chondrocytes was reduced in the presence of salicylate and increased in the presence of misoprostol. When osteoarthritic chondrocytes were incubated with both agents, the salicylate effect was reversed by misoprostol. Collagen synthesis by the chondrocytes from the contralateral knee was also suppressed by salicylate, but addition of misoprostol failed to restore synthesis. In summary, misoprostol, even at very high doses, has limited chondroprotective activity in canine cartilage, as judged from collagen synthetic activity. PMID- 11856994 TI - Misoprostol Inhibits Polymethylmethacrylate-Stimulated Lysosomal Degranulation and IL-1 Release from Neutrophils. AB - Endoprosthetic orthopedic implants may loosen over time. The mechanism of this loosening process remains poorly understood. Wear debris sloughed from bone cement (polymethylmethacrylate, PMMA) and orthopedic implant materials (metal, ultrahigh-molecular-weight polyethylene) may stimulate inflammatory responses in phagocytic cells which populate the bone-implant interface (synovial-like membrane). This investigation aimed to determine whether the prostaglandin-E(1) (PGE(1)) analog misoprostol might modulate PMMA-stimulated phagocytic cell degranulation and the release of interleukins such as IL-1. Lysozyme and IL-1 release from PMMA-stimulated neutrophils in vitro were measured as approximately 0.07 &mgr;g per 10(6) cells per min and 4 pg per 10(6) cells per min, respectively. These rates decreased to 0.03 &mgr;g per 10(6) cells per min and 1.7 pg per 10(6) cells per min, respectively, after the addition of 50 nM misoprostol to the incubation medium. Misoprostol inhibited degranulation and cytokine release in a dose-dependent manner. Consequently, misoprostol modulates PMMA-stimulated inflammatory responses. These responses appear to be mediated by prostanoids, and the regulation of prostanoids at the bone-implant interface may modulate the release of inflammatory osteolytic mediators (PGE(2)) which contribute to implant loosening. PMID- 11856995 TI - Particulate-Induced, Prostaglandin- and Cytokine-Mediated Bone Resorption in an Experimental System and in Failed Joint Replacements. AB - Total hip arthroplasty (THA) has provided dramatic pain relief and improvement in function for millions of patients with end-stage arthritis; however, periprosthetic osteolysis following THA has become increasingly recognized as a major clinical problem in both cemented and cementless reconstructions. An aggressive granulomatous tissue (interfacial membrane) consisting predominantly of fibroblasts, aggregates of macrophages, and foreign body giant cells develops at the interface of bone/prostheses or bone/cement. It is believed that particulate wear debris from prosthetic materials and/or bone cement are phagocytized by histiocytic cells of interfacial membrane and then these cells produce inflammatory mediators and proteolytic enzymes to provoke a cascade of osteolytic events. In this paper, we studied in vitro responsiveness of various cell types to particulate wear debris. Although titanium and titanium alloys demonstrate excellent biocompatibility in bulk from, titanium in particulate form can provoke a variety of cellular responses. We have found that small-sized Ti particles of phagocytosable size, a commonly encountered particle species in the periprosthetic tissues of failed THAs, stimulate macrophages to secrete various mediators of bone resorption (prostaglandin E(2), interleukin-1, interleukin-6, and tumor necrosis factor-alpha from macrophages and cause bone resorption in organ culture. In addition, we have shown that phagocytosable titanium particles stimulate fibroblasts to up-regulate the expression of matrix metalloproteinases (stromelysin and collagenase) without a substantial effect on the tissue inhibitor of these enzymes (TIMP). Titanium particulates also have a suppressive effect on procollagen synthesis by osteoblast-like cell line. Thus, titanium particulates have the capacity to stimulate bone resorption and inhibit bone matrix formation. In this series of experiments, we have also shown in vitro inhibitory effect of certain pharmaceutical components (indomethacin, misoprostol) upon bone resorption in organ culture, which may indicate a potential therapeutic intervention to prevent or treat particulate-induced pathological bone resorption in total joint arthroplasties. PMID- 11856996 TI - Immunoregulation of Proteoglycan-Induced Arthritis in Balb/c Mice. AB - Immunization of BALB/c mice with a select group of cartilage proteoglycans induces progressive polyarthritis. The pathological mechanisms of proteoglycan induced arthritis are based on autoimmune reactions developed against the mouse self-proteoglycan. This autoimmune inflammatory animal model, which shows many similarities to human rheumatoid arthritis, has close to 100% incidence in susceptible BALB/c mice and is an excellent in vivo model for testing disease modifying agents. The aim of this work was to study the regulatory mechanisms which control the autoimmune reactions in proteoglycan-induced arthritis, in association with the release of most important cytokines/mediators (interleukin 1 (IL-1), interleukin-2 [IL-2], interleukin-6 [IL-6], tumor necrosis factor-alpha [TNF-alpha], and prostaglandin E(2)) which are believed to play key roles in inflammatory events and cartilage degredation. We have found relatively high levels of IL-1 sera and synovial-cell culture supernatants of arthritic animals, whereas IL-1 was not detected in nonarthritic control animals. Serum autoantibody and IL-1 levels seemed to be sensitive indicators of the oncoming inflammatory events in the joints, whereas autoreactive T-cell responses to mouse proteoglycan became evident only after the onset of arthritis. As proteoglycan-specific T-cell responses were mainly restricted to the joint-draining lymph nodes during the arthritic process, it is likely that the autoantigen-driven mechanism of joint inflammation became local and self-sustaining during cartilage degradation. Thus, autoimmune mechanisms seem to be essential in the "organ specificity" of inflammatory reactions, as "arthritogenic" lymphocytes migrate to and accumulate in both the lymphoid organs and the synovium. IL-1 and IL-2 are among the most important mediators in proteoglycan-induced arthritis and are able to influence autoimmune reactions and the migration of lymphocytes to the synovium. PMID- 11856997 TI - Perturbation of a Cartilage Autocrine/Paracrine Basic Fibroblast Growth Factor Metabolic Regulatory Network by Osteoarthritic Synovial Tissue. AB - Studies have focused on control of expression and the relative importance of basic fibroblast growth factor (bFGF) in a purported autocrine/paracrine regulatory network functioning in the modulation of cartilage metabolic and structural homeostasis. Preformed and newly synthesized bFGF and concurrent antagonist activity could be identified by bioassay in cell/pericellular matrix extracts of normal bovine articular chondrocytes maintained in suspension culture. Specificity was determined using antibody neutralization. Prostanoids (PGE(1), PGE(2)) enhanced chondrocyte expression of the putative inhibitor. The antagonist, recognized in the presence of suboptimally triggered bFGF receptors, was active against both endogenously produced and recombinant bFGF. Chondrocyte expression of bFGF was significantly altered following exposure to conditioned medium obtained from explant cultures of osteoarthritic synovial tissue. Response pattern, that is, an upregulation or downregulation of growth factor expression, was dependent on medium concentration and the duration of chondrocyte exposure. Synovium-conditioned medium generated in the presence of PGE(1) appeared to attenuate suppressive responses seen with naive conditioned medium. Promotion of expression of bFGF inhibitory activity within the milieu of the diseased joint may negate potential detrimental pathophysiologic effects of this competence factor on cartilage, synovial tissue, and bone metabolism and repair. PMID- 11856998 TI - Central Nervous System Effects of Ginkgo Biloba, a Plant Extract. AB - Extracts of Ginkgo biloba (EGb) are among the most prescribed drugs in France and Germany. EGb is claimed to be effective in peripheral arterial disorders and in "cerebral insufficiency." The mechanism of action is not yet well understood. Three of the ingredients of the extract have been isolated and found to be pharmacologically active, but which one alone or in combination is responsible for clinical effects is unknown. The recommended daily dose (3 x 40 mg extract) is based more on empirical data than on clinical dose-findings studies. However, despite these, according to double-blind, placebo-controlled clinical trials, EGb has therapeutic effects, at least, on the diagnostic entity of "cerebral insufficiency," which is used in Europe as synonymous with early dementia. To determine whether EGb has significant pharmacological effects on the human brain, a pharmacodynamic study was conducted using the Quantitative Pharmacoelectroencephalogram (QPEEG(R)) method. It was established that the pharmacological effects (based on a predetermined 7.5--13.0-Hz alpha frequency band in a computer-analyzed electroencephalogram = CEEG(R)) of EGb on the central nervous system (CNS) are significantly different than placebo, and the high and low doses could be discriminated from each other. The 120-mg, but particularly the 240-mg, single doses showed the most consistent CNS effects with an earlier onset (1 h) and longer duration (7 h). Furthermore, it was established that the electrophysiological effects of EGb in CNS are similar to those of well-known cognitive activators such as "nootropics" as well as tacrine, the only marketed "antidementia" drug currently available in the United States. PMID- 11856999 TI - Isoniazid Acetylation Phenotyping in the Japanese: The Molar Metabolic Ratio INH/AcINH. AB - The N-acetylation phenotyping of isoniazid (INH) was studied in 434 unrelated Japanese pulmonary tuberculosis patients. The frequency of slow acetylators was determined using three methods based on the urinary levels of INH and AcINH: percentage acetylisoniazid (%AcINH), the inactivation index or acetylation index (AcINH/INH), and the molar metabolic ratio (INH/AcINH) in urine. Frequency histograms and probit plots were constructed with the data obtained from each method. Using %AcINH with the conventional antimode of 70%, the number of slow acetylator was 12.7%. Using AcINH/INH, the number of slow acetylator was 52% according to the conventional antimode of 6.0. The molar metabolic ratio INH/AcINH showed explicitly the best bimodality and a clear-cut antimode among these three methods. From probit plots of INH/AcINH, an antimode of 0.45 can be suggested for the 434 Japanese patients; 377 patients (86.9%) as rapid acetylator and 57 patients (13.1%) as slow acetylator. PMID- 11857000 TI - The Pharmacology of Alcohol Withdrawal Syndrome Treatment Reviewed: Efficacy, Cost, and Safety. AB - Alcohol continues to be one of the most common drugs of abuse. The morbidity and mortality associated with alcohol withdrawal has decreased significantly with the advent of pharmacologic intervention. However, the best method for the treatment of alcohol withdrawal syndrome remains controversial. With chronic use, alcohol can disturb the function and balance of several neurotransmitter systems such as excitatory amino acids, GABA, serotonin, and acetylcholine. Compensatory mechanisms in these pathways appear to predominate during chronic use but may become pathologic during withdrawal. As the understanding of alcohol-induced cellular changes increases, treatment of chronic abuse and withdrawal can be refined. Several groups of drugs are efficacious as either primary or adjunct agents in the treatment of alcohol withdrawal. This review describes the current theories on the etiology and treatment of alcohol withdrawal syndrome with emphasis on efficacy, cost, pharmacokinetic parameters, and safety considerations. A proposed withdrawal regimen is also described. Benzodiazepines appear to be the safest and most efficacious choice. With a wide variety of pharmacokinetic parameters and low cost of treatment, they remain the drugs of choice for withdrawal. Ethanol, barbiturates, antiseizure medications, and sympatholytic and sympathomimetic drugs are also reviewed. Newer agents that may have a future role in withdrawal are discussed as well. PMID- 11857002 TI - Weekly platinum chemotherapy for recurrent ovarian cancer. PMID- 11857004 TI - Safe administration of etoposide phosphate after hypersensitivity reaction to intravenous etoposide. AB - Etoposide is commonly used in a variety of malignancies. A well known but rare toxicity are hypersensitivity reactions, usually manifested by chest discomfort, dyspnoea, bronchospasm and hypotension. We report the details of a patient who developed hypersensitivity reactions to intravenous etoposide, but subsequently tolerated the administration of intravenous etoposide phosphate with no sequelae. PMID- 11857003 TI - ONYX-015: mechanisms of action and clinical potential of a replication-selective adenovirus. AB - Accumulated knowledge in the molecular processes of tumour development combined with the availability of genetically modified viruses resemble the basis for new promising cancer therapeutics. The main advantages of employing replication competent viruses are achievement of tumour selective killing and amplification of their oncolytic potential within the tumour mass. In this review, we describe the development of ONYX-015, one of the first and most advanced replication competent viruses for cancer therapy. We discuss the molecular biology of this therapeutic approach and the interesting results obtained with this virus in clinical trials. PMID- 11857005 TI - Virtual brain tumours (gliomas) enhance the reality of medical imaging and highlight inadequacies of current therapy. AB - Gliomas are brain tumours that differ from most other cancers by their diffuse invasion of the surrounding normal tissue and their notorious recurrence following all forms of therapy. We have developed a mathematical model to quantify the spatio-temporal growth and invasion of gliomas in three dimensions throughout a virtual human brain. The model quantifies the extent of tumorous invasion of individual gliomas in three-dimensions to a degree beyond the limits of present medical imaging, including even microscopy, and makes clear why current therapies based on existing imaging techniques are inadequate and cannot be otherwise without other methods for detecting tumour cells in the brain. The model's estimate of the extent of tumourous invasion beyond that defined by standard medical imaging can be useful in more accurately planning therapy regimes as well as predicting sites of potential recurrence without waiting for reemergence on follow-up imaging. PMID- 11857006 TI - Weekly cisplatin and daily oral etoposide is highly effective in platinum pretreated ovarian cancer. AB - We investigated the potential of weekly cisplatin and daily oral etoposide followed by oral etoposide maintenance therapy in patients with platinum refractory ovarium cancer. One hundred and seven patients were entered on the study, 98 patients completed the induction therapy consisting of cisplatin at either 50 or 70 mg m(-2) weekly for six administrations plus oral etoposide at a dose of 50 mg daily. Of these 98 patients, 38 had a platinum treatment-free interval of more than 12 months, 32 had an interval between 4 and 12 months, and 28 had progressed during or within 4 months after last platinum therapy. We assessed response rates and time to progression, and also response duration and survival. Analyses were done on the 98 evaluable patients. All 107 patients were considered evaluable for toxicity. Of the 38 patients with a treatment-free interval of more than 12 months, 92% responded, with 63% complete responses. The median progression-free survival in these patients was 14 months, and the median survival was 26 months. Of the 32 patients with an interval of 4-12 months, 91% responded, with 31% complete responses, a median progression-free interval of 8 and a median overall survival of 16 months. Of the 28 patients with platinum refractory disease, 46% as yet responded, with 29% complete responses, median progression-free interval of 5 and an overall survival of 13 months. Haematologic and non-haematologic, particularly renal toxicity and neurotoxicity, were notably mild. We conclude that this intensive regimen of weekly cisplatin plus daily etoposide is highly effective and well tolerated in patients with ovarian cancer relapsing after conventional platinum-based combination chemotherapy, including patients who have progressed during or within 4 months after platinum treatment. PMID- 11857007 TI - The effect of early pregnancy following chemotherapy on disease relapse and foetal outcome in women treated for gestational trophoblastic tumours. AB - Little literature exists on the safety of early pregnancy following chemotherapy. Here we assess the rate of relapse and foetal outcome in women who have completed single and multi-agent chemotherapy for gestational trophoblastic tumours. The records of 1532 patients treated for persistent gestational trophoblastic tumours at Charing Cross Hospital between 1969 and 1998 were reviewed. Patients were defined as receiving single agent or multi-agent treatment. Relapse rates and foetal outcome were reviewed in the 230 patients who became pregnant within 12 months of completing chemotherapy. In the single agent group 153 (22%) of 691 patients conceived early. Three subsequently relapsed. In the multi-agent group, 77 (10%) of 779 patients conceived early, two then relapsed. Relapse rates were 2% (3 out of 153) and 2.5% (2 out of 77) for each group compared to 5% and 5.6% in the comparative non-pregnant groups. Outcomes of 230 early pregnancies: 164 (71%) delivered at full term, 35 (15%) terminations, 26 (11%) spontaneous abortions, three (1.3%) new hydatidiform moles and two (1%) stillbirths. Early pregnancies were more common in the single agent group (P<0.001), but spontaneous miscarriages and terminations were more likely to occur in the multi-agent group (P=0.04 and 0.03, respectively). Of the full-term pregnancies, three (1.8%) babies were born with congenital abnormalities. Patients in either group who conceive within 12 months of completing chemotherapy are not at increased risk of relapse. Though, we still advise avoiding pregnancy within 12 months of completing chemotherapy, those that do conceive can be reassured of a likely favourable outcome. DOI: 10.1038/sj/bjc/6600041 www.bjcancer.comCopyright 2002 The Cancer Research Campaign PMID- 11857008 TI - Clinicoprognostic implications of increased serum levels of vascular endothelial growth factor and basic fibroblastic growth factor in early B-cell chronic lymphocytic leukaemia. AB - To assess the relative merit of increased serum levels of vascular endothelial growth factor and basic fibroblastic growth factor in predicting the risk of disease progression of patients with early B-cell chronic lymphocytic leukaemia we analyzed 81 Binet stage A patients whose sera were taken at the time of diagnosis and evaluated for the presence of vascular endothelial growth factor and basic fibroblast growth factor using an enzyme-linked immunosorbent assay. Serum levels of vascular endothelial growth factor positively correlated with Rai sub-stages (P=0.03), peripheral blood lymphocytosis (P=0.03), bone marrow histology (P=0.04) and beta2-microglobulin (beta2-m) (P=0.006). When dealing with basic fibroblast growth factor only a correlation with Rai sub-stages (P=0.02) could be found. Different cut-offs set on the basis of a stratification in quartiles, failed to demonstrate any correlation between serum levels of basic fibroblast growth factor and disease progression. In contrast, patients with increased serum levels of vascular endothelial growth factor (above median value, 203 pg ml(-1)) had a three times increased risk of disease progression, although, in multivariate analysis only Rai sub-stages (P=0.0001) and lymphocyte doubling time (P=0.002) retained their prognostic significance. Low levels of vascular endothelial growth factor were indicative of good clinical outcome in the subgroup of patients with either low (P=0.02) or high (P=0.03) beta2-m concentration. Finally, the highest prognostic power was obtained when serum vascular endothelial growth factor and beta2-m were examined in combination. Median of progression-free survival of patients who had both serum vascular endothelial growth factor and beta2-m higher than median value was only 13 months, in contrast median progression-free survival of patients with one marker increased (i.e. above the 50th percentile) was 40 months. Patients with both markers below the median experienced the best clinical outcome (median progression-free survival not reached at 40 months). In conclusion, serum levels of either vascular endothelial growth factor or basic fibroblast growth factor are high in patients with early chronic lymphocytic leukaemia, however, only vascular endothelial growth factor predicts behaviour of disease and helps to refine the prognosis of stage A patients. PMID- 11857009 TI - Lung cancer referral patterns in the former Yorkshire region of the UK. AB - The purpose of this study was to find out what proportion of patients are referred as lung cancer guidelines assume, whether different referral pathways result in different management and what proportion of patients are seen within recommended time intervals between referral and treatment. A randomly selected sample of 400 lung cancer cases registered with the former Yorkshire Cancer Registry database in 1993 was selected for casenote analysis. Mode of presentation, speciality of initial referral, treatment by specialist, time intervals for key points in the referral pathways were analyzed. A total of 362 (90.5%) of case-notes were available. Less than half of lung cancer patients (173, 47.8%) presented to hospital with a chest X-ray diagnosis of lung cancer. Forty-one (11.3%) presented as self-referrals to Accident and Emergency and the remainder were referred without a diagnosis of lung cancer by other routes, mainly via GPs. Patients who did not present initially with a lung cancer diagnosis were less likely to receive specialist care (62%:96%), or have their diagnosis histologically confirmed (57.1%:80.3%) or receive surgery or radical radiotherapy (6.9%:13.9%). Nine per cent of all 362 patients did not receive a specialist opinion. Eighty per cent of patients referred by a GP with CXR suspected lung cancer were seen at hospital within 2 weeks. Only 32.4% of those receiving active treatment were treated within 8 weeks of clinical diagnosis or first hospital visit. Lung cancer patients presenting to hospital without a suspicious CXR are less likely to have specialist care, histological confirmation of their cancer and have lower rates of active treatment (surgery, any radiotherapy or chemotherapy). PMID- 11857010 TI - Changes in psychological distress after cancer genetic counselling: a comparison of affected and unaffected women. AB - This study sought to examine changes in psychological distress following cancer genetic counselling. Women attending a family cancer clinic completed questionnaires before their appointment and at 2 weeks, 6 months and 12 months after their appointment. Twenty-six women were at low risk of developing breast or ovarian cancer, 76 were at moderate risk, 46 were at high risk and 46 women had previously had breast or ovarian cancer. All groups were compared with regard to measures of anxiety, depression, general psychological distress, worry about developing breast and ovarian cancer, and perceived risk of developing breast/ovarian cancer and perceived likelihood of carrying a genetic mutation. General psychological distress did not change over the course of the study and the groups did not differ on these measures. Worry about developing breast cancer and perceptions of the likelihood of carrying a genetic mutation significantly reduced following genetic counselling. On the whole women who had already had breast/ovarian cancer showed more concerns about ovarian cancer and raised perceptions of risk in comparison with the other groups, indicating the need for sensitive counselling of such women. PMID- 11857011 TI - Computer-based quality of life questionnaires may contribute to doctor-patient interactions in oncology. AB - It is well recognized that oncologists should consider patients' quality of life and functioning when planning and delivering anticancer treatment, but a comprehensive assessment of how a patient feels requires a thorough inquiry. A standardized measurement of patients' quality of life may support clinicians in identifying important problems for discussion during the limited time of the medical consultations. The aim of this study was to assess the feasibility of computer-administered individual quality of life measurements in oncology clinics with immediate feedback of results to clinicians and to examine the impact of the information on consultations. The study employed a prospective non-randomized design with pre-test post-test within subjects comparisons and involved three medical oncologists and 28 cancer patients receiving chemotherapy. The intervention consisted of completion of quality of life questionnaires before the consultations and informing clinicians of the results. The main outcome measures were patients' perceptions of the content of baseline and intervention consultations and satisfaction with communication. A qualitative analysis of clinicians' interviews was performed. When clinicians had the quality of life results they enquired more often about daily activities (Z= -2.71, P=0.007), emotional problems (Z= -2.11, P=0.035) and work related issues (Z= -1.89, P=0.058). There was an increase in the number of issues discussed during the intervention consultation (Z= -1.89, P=0.059). Patients were highly satisfied with both consultations. The computer measurement was well accepted by patients who felt that the questionnaires were a useful tool to tell the doctors about their problems. The clinicians perceived that the quality of life data broadened the range of the clinical inquiry and helped them identify issues for discussion. Having symptoms and functional problems expressed quantitatively on a scale was useful for detection of change over time. PMID- 11857012 TI - Smoking and the risk of acute myeloid leukaemia in cytogenetic subgroups. AB - Cytogenetically-defined subgroups of acute myeloid leukaemia have distinct biologies, clinical features and outcomes. Evidence from therapy-related leukaemia suggests that chromosomal abnormalities are also markers of exposure. Our results suggest that the smoking-associated risk for acute myeloid leukaemia is restricted to the t(8;21)(q22;q22) subgroup. This supports the hypothesis that distinct cytogenetic subgroups of acute myeloid leukaemia have separate aetiologies. PMID- 11857013 TI - Vitamin K and childhood cancer: analysis of individual patient data from six case control studies. AB - To investigate the hypothesis that neonates who receive intramuscular vitamin K are at an increased risk of developing cancer, particularly leukaemia, a pooled analysis of individual patient data from six case-control studies conducted in Great Britain and Germany has been undertaken. Subjects comprised 2431 case children diagnosed with cancer before 15 years of age and 6338 control children. The retrospective assessment of whether or not an individual baby received vitamin K is not straightforward. In many cases no record was found in stored medical notes and two types of analysis were therefore conducted; in the first it was assumed that where no written record of vitamin K was found it had not been given, and in the second, where no written record of administration was found, information on hospital policy and perinatal morbidity was used to 'impute' whether or not vitamin K had been given. In the first analysis, no association was found between neonatal administration of intramuscular. vitamin K and childhood cancer: odds ratios adjusted for mode of delivery, admission to special care baby unit and low birth weight were 1.09 (95% confidence interval 0.92-1.28) for leukaemia and 1.05 (0.92-1.20) for other cancers. In the second analysis, the adjusted odds ratios increased to 1.21 (1.02-1.44) for leukaemia and 1.10 (0.95 1.26) for other cancers. This shift did not occur in all studies, and when data from the hypothesis generating Bristol study were excluded, the adjusted odds ratios for leukaemia became 1.06 (0.89-1.25) in the first analysis and 1.16 (0.97 1.39) when data on prophylaxis imputed from hospital policy and perinatal morbidity were used. We conclude that whilst the broad nature of the diagnostic groups and the poor quality of some of the vitamin K data mean that small effects cannot be entirely ruled out, our analysis provides no convincing evidence that intramuscular vitamin K is associated with childhood leukaemia. PMID- 11857014 TI - Occupational physical activity and risk for prostate cancer in a nationwide cohort study in Sweden. AB - We investigated effects of occupational physical activity on relative risk for prostate cancer. From Swedish nationwide censuses in 1960 and 1970, we defined two cohorts of men whose occupational titles allowed classification of physical activity levels at work in 1960 (n=1,348,971) and in 1970 (n=1,377,629). A third cohort included only men whose jobs required a similar level of physical activity in both 1960 and 1970 (n=673,443). The incidence of prostate cancer between 1971 and 1989 was ascertained through record linkage to the Swedish Cancer Register. A total of 43,836, 28,702, and 19,670 prostate cancers, respectively, occurred in the three cohorts. In all three cohorts, the relative risk for prostate cancer increased with decreasing level of occupational physical activity (P<0.001), using Poisson regression. Among men with the same physical activity levels in 1960 and 1970, the rate ratio was 1.11 for men with sedentary jobs as compared with those whose jobs had very high/high activity levels after adjustment for age at follow-up, calendar year of follow-up and place of residence (95% CI 1.05 1.17; P for trend <0.001). There was no association between occupational activity and prostate cancer mortality. Since we had no data on other potential risk factors the observed associations for both incidence and mortality might have been confounded. Further studies are needed to better understand the potential role of physical activity for prostate cancer. PMID- 11857015 TI - A comprehensive model for familial breast cancer incorporating BRCA1, BRCA2 and other genes. AB - In computing the probability that a woman is a BRCA1 or BRCA2 carrier for genetic counselling purposes, it is important to allow for the fact that other breast cancer susceptibility genes may exist. We used data from both a population based series of breast cancer cases and high risk families in the UK, with information on BRCA1 and BRCA2 mutation status, to investigate the genetic models that can best explain familial breast cancer outside BRCA1 and BRCA2 families. We also evaluated the evidence for risk modifiers in BRCA1 and BRCA2 carriers. We estimated the simultaneous effects of BRCA1, BRCA2, a third hypothetical gene 'BRCA3', and a polygenic effect using segregation analysis. The hypergeometric polygenic model was used to approximate polygenic inheritance and the effect of risk modifiers. BRCA1 and BRCA2 could not explain all the observed familial clustering. The best fitting model for the residual familial breast cancer was the polygenic, although a model with a single recessive allele produced a similar fit. There was also significant evidence for a modifying effect of other genes on the risks of breast cancer in BRCA1 and BRCA2 mutation carriers. Under this model, the frequency of BRCA1 was estimated to be 0.051% (95% CI: 0.021-0.125%) and of BRCA2 0.068% (95% CI: 0.033-0.141%). The breast cancer risk by age 70 years, based on the average incidence over all modifiers was estimated to be 35.3% for BRCA1 and 50.3% for BRCA2. The corresponding ovarian cancer risks were 25.9% for BRCA1 and 9.1% for BRCA2. The findings suggest that several common, low penetrance genes with multiplicative effects on risk may account for the residual non-BRCA1/2 familial aggregation of breast cancer. The modifying effect may explain the previously reported differences between population based estimates for BRCA1/2 penetrance and estimates based on high-risk families. PMID- 11857017 TI - Birth weight as a predictor of breast cancer: a case-control study in Norway. AB - The hypothesis that birth weight is positively associated with adult risk of breast cancer implies that factors related to intrauterine growth may be important for the development of this malignancy. Using stored birth records from the two main hospitals in Trondheim and Bergen, Norway, we collected information on birth weight, birth length and placenta weight among 373 women who developed breast cancer. From the same archives, we selected as controls 1150 women of identical age as the cases without a history of breast cancer. Information on age at first birth and parity were collected from the Central Person Registry in Norway. Based on conditional logistic regression analysis, breast cancer risk was positively associated with birth weight and with birth length (P for trend=0.02). Birth weights in the highest quartile (3730 g or more) were associated with 40% higher risk (odds ratio, 1.4, 95% confidence interval, 1.1-1.9) of breast cancer compared to birth weights in the lowest quartile (less than 3090 g). For birth length, the odds ratio for women who were 51.5 cm or more (highest quartile) was 1.3 (95% confidence interval, 1.0-1.8) compared to being less than 50 cm (lowest quartile) at birth. Adjustment for age at first birth and parity did not change these estimates. Placenta weight was not associated with breast cancer risk. This study provides strong evidence that intrauterine factors may influence future risk of breast cancer. A common feature of such factors would be their ability to stimulate foetal growth and, simultaneously, to influence intrauterine development of the mammary gland. PMID- 11857016 TI - Weight at birth and adolescence and premenopausal breast cancer risk in a low risk population. AB - We assessed breast cancer risk in relation to weight at birth and adolescence. In person interviews were completed with the biological mothers of women aged 45 years and younger who participated in the Shanghai Breast Cancer Study in 1996-98 (288 cases, 350 controls). After adjustment for confounding, women who were 4000 g or more at birth were not at increased risk of breast cancer (odds ratio=0.7; 95% confidence interval 0.4-1.4) relative to women whose birth weight was 2500 2999 g. Compared with women of average perceived weight at age 15 years, no relation was apparent for heavier than average weight based on maternal report (odds ratio=0.7; 95% confidence interval 0.5-1.2) or self-report (odds ratio=1.0; 95% confidence interval 0.7-1.6). Perceived adolescent weight and height did not modify the association of birth weight with breast cancer risk. These results suggest that weight early in life is not related to premenopausal breast cancer risk in this low-risk population. PMID- 11857018 TI - The effects of tricyclic antidepressants on breast cancer risk. AB - To test the hypothesis that tricyclic antidepressant use increases invasive female breast cancer incidence, we carried out a case-control study within the population of female beneficiaries of the Saskatchewan Prescription Drug Plan aged 35 years from 1981-995 with no history of cancer since 1970. This agency has provided full or partial coverage for outpatient prescriptions to Saskatchewan residents since 1975. We accrued 5882 histologically proven cases and 23,517 controls, randomly selected from the source population and individually matched on age and sampling time. Heavy exposure to any tricyclic antidepressants was associated with an elevated rate ratio for breast cancer 11-15 years later (2.02, 95% confidence interval: 1.34-3.04). Post hoc analyses based on the results of genotoxicity studies carried out using Drosophila melanogaster suggested that the increased risk could be attributed to the use of the six genotoxic tricyclic antidepressants, and not to the use of the four nongenotoxic tricyclic antidepressants. However, our results may have been confounded by the effects of other determinants of breast cancer associated with tricyclic antidepressant use. PMID- 11857019 TI - Inverse correlation between E-cadherin and Snail expression in hepatocellular carcinoma cell lines in vitro and in vivo. AB - Hepatocellular carcinoma is a well-known malignancy in the world. However, the molecular mechanism of carcinogenesis and tumour progression remains unclear. Recently, reduced E-cadherin expression due to transcriptional suppressor Snail was proven in a panel of epithelial and dedifferentiated cells derived from carcinomas of various etiologies. In the present study, we examined Snail and E cadherin mRNA/protein expression in five hepatocellular carcinoma cell lines with variable phenotypes (HuL-1, Hep-G(2), Changliver, HLE, and HLF). The results demonstrated that the presence of Snail mRNA in HuL-1, Changliver, HLE and HLF cells detected by RT-PCR, which was further proven by in situ hybridization in tumours induced by HuL-1, Changliver, and HLF cells where Snail mRNA signals expressed in each of the sections. By contrast, E-cadherin mRNA and protein expression were only detected in Hep-G(2) cells by RT-PCR and Western blot, respectively. These results were also consistent with the data obtained from in vivo immunohistochemical staining where membranous expression of endogenous E cadherin protein was revealed only in tumour sections induced by Hep-G(2) cells. Here we are the first to report that there is an inverse correlation between Snail and E-cadherin expression in HCC cells as well. PMID- 11857020 TI - Comparison of the RNA-amplification based methods RT-PCR and NASBA for the detection of circulating tumour cells. AB - Increasingly, reverse transcriptase polymerase chain reaction (RT-PCR) is used to detect clinically significant tumour cells in blood or bone marrow. This may result in a redefinition of disease-free and clinical relapse. However, its clinical utility may be limited by lack of automation or reproducibility. Recent studies have suggested nucleic acid sequence-based amplification of target RNA may be more robust. In this study, nucleic acid sequence-based amplification was established to detect melanoma, colorectal and prostate cancer cells. Nucleic acid sequence-based amplification and RT-PCR both successfully amplified target RNA in peripheral blood samples from patients with melanoma and colorectal cancer, but only RT-PCR detected PSA in blood samples from patients with prostate cancer. There was relatively good agreement between sample replicates analyzed by RT-PCR (Kappa values of one for tyrosinase, 0.67 for CK-20 and one for PSA), but less agreement when analyzed by nucleic acid sequence-based amplification. This may limit the routine use of NASBA for the detection of clinically significant disease. In summary, RT-PCR appears at present to be the most reliable and reproducible method for the detection of low-level disease in cancer patients, although prospective studies are warranted to assess the clinical utility of different molecular diagnostic methods. PMID- 11857021 TI - Evaluation of MAGE-1 and MAGE-3 as tumour-specific markers to detect blood dissemination of hepatocellular carcinoma cells. AB - The members of MAGE gene family are highly expressed in human hepatocellular carcinoma (HCC). In the present study, we tested the tumour-specific MAGE-1 and MAGE-3 transcripts in the peripheral blood of HCC patients by nested RT-PCR to detect the circulating tumour cells and evaluate their potential clinical implication. Of 30 HCC patients, the positive rate of MAGE-1 and MAGE-3 transcripts was 43.3% (13 out of 30) and 33.3% (10 out of 30) in PBMC samples, whilst the positive rate was 70% (21 out of 30) and 53.3% (16 out of 30) in the resected HCC tissue samples, respectively. The positivity for at least one MAGE gene transcript was 63.3% (19 out of 30) in PBMC samples of HCC patients and 83.3% (25 out of 30) in the resected HCC tissue samples. MAGE-1 and/or MAGE-3 mRNA were not detected in the PBMC of those patients from whom the resected HCC tissues were MAGE-1 or MAGE-3 mRNA negative, nor in the 25 PBMC samples from healthy donors. The detection of MAGE transcripts in PBMC was correlated with the advanced stages and tumour size of the HCC, being 82.4% (14 out of 17) in tumour stages III and IVa, 56.6% (five out of nine) in stage II, and null (nought out of four) in stage I. The serum alpha-FP in 33.3% (10 out of 30) of HCC patients was normal or slightly elevated (< 40 ng ml(-1)). However, six of these 10 patients (alpha-FP < 40 ng ml(-1)) were MAGE-1 and /or MAGE-3 mRNA positive in their PBMC. The follow-up survey of MAGE mRNA in PBMC was performed in 12 patients. Seven patients with persistent MAGE-1 and/or MAGE-3 mRNA positive or from negative turned to positive died because of metastasis and/or recurrence. In striking contrast, all four patients with MAGE-1 and/or MAGE-3 mRNA from positive turned to negative and one patient with persistent MAGE-3 transcript negative are alive after last test. Collectively, detection of MAGE transcripts with follow-up survey in PBMC is a feasible and reliable assay for the early prediction of the relapse and prognosis of the HCC patients. PMID- 11857022 TI - Galanin and galanin receptor expression in neuroblastic tumours: correlation with their differentiation status. AB - Neuroblastoma and its benign differentiated counterpart, ganglioneuroma, are paediatric neuroblastic tumours arising in the sympathetic nervous system. Their broad spectrum of clinical virulence is mainly related to heterogeneous biologic background and tumour differentiation. Neuroblastic tumours synthesize various neuropeptides acting as neuromodulators. Previous studies suggested that galanin plays a role in sympathetic tissue where it could be involved in differentiation and development. We investigated the expression and distribution of galanin and its three known receptors (Gal-R1, Gal-R2, Gal-R3) in 19 samples of neuroblastic tumours tissue by immunohistochemistry, in situ hybridization and fluorescent ligand binding. This study provides clear evidence for galanin and galanin receptor expression in human neuroblastic tumours. The messengers coding for galanin, Gal-R1 and -R3 were highly expressed in neuroblastoma and their amount dramatically decreased in ganglioneuroma. In contrast, Gal-R2 levels remained unchanged. Double labelling studies showed that galanin was mainly co-expressed with its receptors whatever the differentiation stage. In neuroblastic tumours, galanin might promote cell-survival or counteract neuronal differentiation through the different signalling pathways mediated by galanin receptors. Finally, our results suggest that galanin influences neuroblastoma growth and development as an autocrine/paracrine modulator. These findings suggest potential critical implications for galanin in neuroblastic tumours development. PMID- 11857023 TI - Effect of manipulation of primary tumour vascularity on metastasis in an adenocarcinoma model. AB - One explanation for the clinical association between tumour vascularity and probability of metastasis is that increased primary tumour vascularity enhances haematogenous dissemination by offering greater opportunity for tumour cell invasion into the circulation (intravasation). We devised an experimental tumour metastasis model that allowed manipulation of primary tumour vascularity with differential exposure of the primary and metastatic tumour site to angiogenic agents. We used this model to assess the effects of local and systemic increases in the level of the angiogenic agent basic fibroblast growth factor on metastasis. BDIX rats with implanted hind limb K12/TR adenocarcinoma tumours received either intratumoural or systemic, basic fibroblast growth factor or saline infusion. Both intratumoural and systemic basic fibroblast growth factor infusion resulted in significant increases in tumour vascularity, blood flow and growth, but not lung metastasis, compared with saline-infused controls. Raised basic fibroblast growth factor levels and increase in primary tumour vascularity did not increase metastasis. The clinical association between tumour vascularity and metastasis is most likely to arise from a metastatic tumour genotype that links increased tumour vascularity with greater metastatic potential. PMID- 11857024 TI - Induction of DNA breaks and apoptosis in crosslink-hypersensitive V79 cells by the cytostatic drug beta-D-glucosyl-ifosfamide mustard. AB - To study molecular aspects of cytotoxicity of the anticancer drug beta-D-glucose ifosfamide mustard we investigated the potential of the agent to induce apoptosis and DNA breakage. Since beta-D-glucose-ifosfamide mustard generates DNA interstrand crosslinks, we used as an in vitro model system a pair of isogenic Chinese hamster V79 cells differing in their sensitivity to crosslinking agents. CL-V5B cells are dramatically more sensitive (30-fold based on D(10) values) to the cytotoxic effects of beta-D-glucose-ifosfamide mustard as compared to parental V79B cells. After 48 h of pulse-treatment with the agent, sensitive cells but not the resistant parental line undergo apoptosis and necrosis, with apoptosis being the predominant form of cell death (70 and 20% of apoptosis and necrosis, respectively). Apoptosis increased as a function of dose and was accompanied by induction of DNA double-strand breaks in the hypersensitive cells. Furthermore, a strong decline in the level of Bcl-2 protein and activation of caspases-3, -8 and -9 were observed. The resistant parental cells were refractory to all these parameters. Bcl-2 decline in the sensitive cells preceded apoptosis, and transfection-mediated overexpression of Bcl-2 protected at least in part from apoptosis. From the data we hypothesize that non-repaired crosslinks induced by beta-D-glucose-ifosfamide mustard are transformed into double-strand breaks which trigger apoptosis via a Bcl-2 dependent pathway. PMID- 11857025 TI - 17beta-Oestradiol treatment modulates nitric oxide synthase activity in MDA231 tumour with implications on growth and radiation response. AB - The putative oestrogen receptor negative human breast cancer cell line MDA231, when grown as tumours in mice continually receiving 17beta-oestradiol, showed substantially increased growth rate when compared to control animals. Further, we observed that 17beta-oestradiol treatment could both increase the growth rate of established MDA231 tumours as well as decreasing the time taken for initiating tumour growth. We have also demonstrated that this increase in growth rate is accompanied by a four-fold increase in nitric oxide synthase activity, which was predominantly the inducible form. Inducible-nitric oxide synthase expression in these tumours was confirmed by immunohistochemical analysis and appeared localized primarily in areas between viable and necrotic regions of the tumour (an area that is presumably hypoxic). Prophylactic treatment with the nitric oxide synthase inhibitor nitro-L-arginine methyl ester resulted in significant reduction in this apparent 17beta-oestradiol-mediated growth promoting effect. Tumours derived from mice receiving 17beta-oestradiol-treatment were characterized by a significantly lower fraction of perfused blood vessels and an indication of an increased hypoxic fraction. Consistent with these observations, 17beta-oestradiol-treated tumours were less radio-responsive compared to control tumours when treated with a single radiation dose of 15 Gy. Our data suggests that long-term treatment with oestrogen could significantly alter the tumour oxygenation status during breast tumour progression, thus affecting response to radiotherapy. PMID- 11857026 TI - Characterization of cell death induced by vinflunine, the most recent Vinca alkaloid in clinical development. AB - Vinflunine, the most recent Vinca alkaloid in clinical development, demonstrated superior antitumour activity to other Vincas in preclinical tumour models. This study aimed to define its molecular mechanisms of cell killing in both parental sensitive and vinflunine-resistant P388 leukaemia cells. Vinflunine treatment of these cells resulted in apoptosis characterized by DNA fragmentation and proteolytic cleavage of poly-(ADP-ribose) polymerase. Apoptosis-inducing concentrations of vinflunine caused c-Jun N-terminal kinase 1 stimulation, as well as caspases-3/7 activation. This activation of caspases and the induction of apoptosis could be inhibited by the caspase inhibitor acetyl-Asp-Glu-Val-Asp aldehyde. Interestingly, the apoptosis signal triggered by vinflunine in these P388 cells was not mediated through Bcl-2 phosphorylation. In addition, when vinflunine resistance was developed in P388 cells, it was associated with resistance to vinflunine-induced apoptosis, as reflected by a loss of capacity to induce DNA fragmentation and PARP degradation, and characterized by increased levels of Bcl-2 and Bfl-1/A1. Therefore, these data indirectly implicate Bcl-2 and Bfl-1/A1 in vinflunine-induced cell death mechanisms. PMID- 11857027 TI - Induction of tumour-specific CD8(+) cytotoxic T lymphocytes by tumour lysate pulsed autologous dendritic cells in patients with uterine serous papillary cancer. AB - Uterine serous papillary carcinoma is a highly aggressive variant of endometrial cancer histologically similar to high grade ovarian cancer. Unlike ovarian cancer, however, it is a chemoresistant disease from onset, with responses to combined cisplatinum-based chemotherapy in the order of 20% and an extremely poor prognosis. In this study, we demonstrate that tumour lysate-pulsed autologous dendritic cells can elicit a specific CD8(+) cytotoxic T lymphocyte response against autologous tumour target cells in three patients with uterine serous papillary cancer. CTL from patients 1 and 2 expressed strong cytolytic activity against autologous tumour cells, did not lyse autologous lymphoblasts or autologous EBV-transformed cell lines, and were variably cytotoxic against the NK sensitive cell line K-562. Patient 3 CD8(+) T cells expressed a modest but reproducible cytotoxicity against autologous tumour cells only at the time of the first priming. Further priming attempts with PBL collected from patient 3 after tumour progression in the lumboaortic lymph nodes were unsuccessful. Cytotoxicity against autologous tumour cells could be significantly inhibited by anti-HLA class I (W6/32) and anti-LFA-1 MAbs. Highly cytotoxic CD8(+) T cells from patients 1 and 2 showed a heterogeneous CD56 expression while CD56 was not expressed by non-cytotoxic CD8(+) T cells from patient 3. Using two colour flow cytometric analysis of intracellular cytokine expression at the single cell level, a striking dominance of IFN-gamma expressors was detectable in CTL populations of patients 1 and 2 while in patient 3 a dominant population of CD8(+) T cells expressing IL-4 and IL-10 was consistently detected. Taken together, these data demonstrate that tumour lysate-pulsed DC can be an effective tool in inducing uterine serous papillary cancer-specific CD8(+) CTL able to kill autologous tumour cells in vitro. However, high levels of tumour specific tolerance in some patients may impose a significant barrier to therapeutic vaccination. These results may have important implications for the treatment in the adjuvant setting of uterine serous papillary cancer patients with active or adoptive immunotherapy. PMID- 11857028 TI - Inhibition of LNCaP prostate tumor growth in vivo by an antisense oligonucleotide directed against the human androgen receptor. AB - We have shown recently that a 15-mer phosphorothioate oligodeoxynucleotide (ODNas750/15) that hybridizes to the (CAG)n polyglutamine region of mRNA encoding human androgen receptor (AR) inhibits the expression of AR in LNCaP prostate cancer cells in vitro. This AR downregulation was accompanied by significant cell growth inhibition and reduced PSA secretion. In the present study we investigated the effects of this antisense AR ODN on prostate tumor growth in vivo using a mouse xenograft model. Via subcutaneously implanted diffusion pumps, either ODNas750/15 or a scrambled control sequence ODNsr750/15 was continuously administered into LNCaP tumor-bearing male nude mice for 7 weeks. Compared with untreated control animals, treatment with ODNas750/15 resulted in significant tumor growth inhibition. Retardation of tumor growth was also significant in castrated mice, whereas the scrambled control ODN did not exert any effects. No side effects such as loss of body weight were observed at any time of treatment. ODN treatment was well tolerated and, in contrast to castration, did not induce shrinkage of mouse prostates. Both AR expression in the tumor and PSA levels in mouse serum correlated with tumor size. However, we failed to demonstrate a correlation between tumor retardation and Ki-67 antigen expression and the number of apoptotic cells, respectively. Testing of antisense-treated LNCaP cells revealed that expression levels of other proteins that contain shorter polyglutamine sequence stretches such as HDAC2, TFIID, and c-jun were not affected. The present study demonstrates that downregulation of AR with antisense ODNas750/15 causes prostate tumor growth inhibition. These results further point out the important role of the AR in prostate tumors and support further testing of AR downregulation for treatment of prostate cancer. PMID- 11857029 TI - Gene therapy of melanoma pulmonary metastasis by intramuscular injection of plasmid DNA encoding tissue inhibitor of metalloproteinases-1. AB - Tumor cell invasion and metastasis are a complex multistep process that involves the degradation of extracellular matrix proteins by matrix metalloproteinases. Tissue inhibitor of metalloproteinase-1 (TIMP-1) acts as a negative regulator of matrix metalloproteinases and thus prevents tumor cell invasion and metastasis by preserving extracellular matrix integrity. In the present study, we investigated whether increasing serum TIMP-1 levels by gene transfer would decrease experimental pulmonary metastasis of melanoma in C57BL/6 mice. Female animals bearing B16F10 melanoma pulmonary metastasis were injected intramuscularly twice per week with 100 microg of plasmid DNA encoding the human TIMP-1 cDNA (TIMP 1pDNA). Substantive levels of serum human TIMP-1 were observed 3 days after single injection and were found for 6 days thereafter. Pulmonary metastasis was significantly reduced in the mice following 4 weeks of TIMP-1 treatment as compared to the controls that were treated with the plasmid DNA vector alone. Further reduction of pulmonary metastasis and increase in survival were realized by intraperitoneal injection of 1000 U of IL-2 twice per week in combination with TIMP-1 treatment. In a parallel in vitro study, a 3-fold increase in TIMP-1 expression was observed in NIH3T3 cells after IL-2 treatment. Therefore, up regulation of TIMP-1 expression by IL-2 likely contributed to the additive effect of IL-2 and TIMP-1 in reducing metastatic disease in the animal model. In conclusion, our findings support the potential of TIMP-1 gene therapy for the prevention of metastatic melanoma. PMID- 11857030 TI - Alternative splicing as a novel of means of regulating the expression of therapeutic genes. AB - In order to determine the potential of alternative splicing as a means of targeting the expression of therapeutic genes to tumor cells in vivo, a series of episomal plasmid-based "splice-activated gene expression" (pSAGE) vectors was generated, which contain minigene cassettes composed of various combinations of the three alternatively spliced exons present in the differentially expressed adhesion protein CD44R1 (v8, v9, and v10) with or without their corresponding intronic sequences, positioned in-frame between the CD44 leader sequence and a "leaderless" human liver/bone/kidney alkaline phosphatase (ALP) cDNA. Because both the v8-v9 and v9-v10 introns contain multiple in-frame stop codons, the expression and enzymatic activity of ALP are dependent upon the accurate removal of intronic sequences from the pre-mRNA transcripts encoded by these constructs. The various pSAGE constructs were introduced into CD44H-positive (T24) and CD44R1 positive (PC3) target cells by electroporation and transfectants selected in hygromycin B. ALP expression was determined by staining with the ALP substrate, BCIP/INT, and the transfected cells tested for their sensitivity to the inactive prodrug, etoposide phosphate. ALP-mediated dephosphorylation of etoposide phosphate generates the potent topoisomerase II inhibitor etoposide. The data obtained indicate that whereas the v8-v9 intron is spliced in both CD44H- and CD44R1-positive cells, the v9-v10 intron is efficiently and accurately removed only in CD44R1-positive cells. Furthermore, only CD44R1-positive cells were sensitized to etoposide phosphate when transfected with the v9-v10.ALP construct. These data emphasize the potential usefulness of alternative splicing as a novel means of targeting gene expression to tumor cells in vivo. PMID- 11857031 TI - In situ cancer vaccination with a replication-conditional HSV for the treatment of liver metastasis of colon cancer. AB - In this study, we investigated the therapeutic efficacy of a replication conditional mutant HSV, G207, for the treatment of liver metastasis of colon carcinoma. Three liver metastasis models in syngeneic BALB/c mice were developed: (i) splenic injection, (ii) splenic and subcutaneous (s.c.) injection, and (iii) orthotopic implantation of CT26 colon carcinoma. In the splenic injection model, G207 was injected into the established splenic tumor on day 7. In the splenic and s.c. injection model, G207 were injected into the established s.c. tumor on days 5 and 8. In the orthotopic implantation model, a piece of CT26 tumor tissue was transplanted onto the wall of the cecum and G207 was injected in the established cecum tumor on day 7. On day 21 or 28, animals were sacrificed and liver metastases were evaluated. In all three models in immunocompetent mice, liver metastases were significantly reduced by intratumoral inoculation with G207 compared to the control. In athymic mice, however, there was no significant therapeutic effect of intratumoral inoculation with G207 on liver metastases. Tumor-specific cytotoxic T-lymphocyte responses were induced in mice treated with G207 in the orthotopic implantation model. These results suggest that intratumoral inoculation of G207, as an in situ cancer vaccine, can be an effective approach against liver metastasis of colon cancer and the efficacy involves tumor-specific T-cell responses. PMID- 11857032 TI - Connexin 43-mediated bystander effect in two rat glioma cell models. AB - In tumor models, the killing by ganciclovir of a fraction of tumor cells transfected with the thymidine kinase (TK) gene has been shown to induce complete regression of the tumor. The mechanism responsible for this bystander effect is thought to be the diffusion of toxic metabolites or apoptotic signals across gap junctions. Connexin 43 (Cx43) is the major component of astrocyte gap junctions. We investigated the susceptibility of two rat glioma cell lines (CNS1 and C6) to thymidine kinase/ganciclovir, before and after transfection with the Cx43 gene. We report a close correlation between the level of Cx43 expression, the extent of gap junctional communication and the amplitude of the bystander effect. Transfection of C6 cells (which display a weak bystander effect and low levels of connexin) with a Cx43 construct induced a strong bystander effect. Inhibition of gap junction activity by 18-alpha-glycyrrhetinic acid abolished the metabolic interaction between TK(+) and TK(-) cells. This metabolic interaction was also abolished if TK(+) and TK(-) cells were separated by a semipermeable membrane. Surprisingly, the transfection of only one of these two interacting cell types with the Cx43 gene was sufficient to induce a bystander effect, although this effect was weaker than that observed if both TK(+) and TK(-) cells expressed Cx43. Finally, Cx43 expression increased sensitivity to contact inhibition. Overall, our data provide evidence that the restoration of gap junctional communication may potentiate HSV/tk-based cancer treatment and suggest that this strategy may have wider application in cancer therapy. PMID- 11857033 TI - A convenient cancer vaccine therapy with in vivo transfer of interleukin 12 expression plasmid using gene gun technology after priming with irradiated carcinoma cells. AB - We studied interleukin (IL)-12 gene therapy using a gene gun as a new autologous vaccination strategy for cancer. In the first experiment, BALB/c mice were inoculated with syngeneic murine renal cancer cells (Renca) intradermally in the abdomen. This was followed by an injection of IL-12 expression plasmid using the gene gun. About 40% of the mice exhibited rejection of the tumor after the treatment and these mice also acquired immunological resistance against a secondary challenge with Renca cells. Based on these results, we examined whether antitumor activity can be potentiated when mice undergo combination treatment with intradermal inoculation of irradiated Renca cells and transfection with IL 12 gene. Inoculation of irradiated Renca cells alone was partially effective in inducing antitumor immunity, whereas the combined treatment remarkably intensified this effect. Moreover, this combined treatment inhibited tumor establishment and enhanced survival of the mice with tumor infiltration by CD4(+) and CD8(+) T cells, even when the treatment was started after tumor-implantation at a distant site. This antitumor effect was antigen specific and we confirmed the induction of antitumor cytotoxic T cells by this treatment. These results show that local cutaneous transfer of IL-12 expression plasmid using gene gun technology enhances systemic and specific antitumor immunity primed by irradiated tumor cells. PMID- 11857035 TI - Gene transfection by quantitatively reconstituted Sendai envelope proteins into liposomes. AB - Fusogenic liposomes (virosomes) consisting of Sendai virus envelope proteins have been utilized for in vitro and in vivo genetic modification of animal cells. In this study, the virosomes containing DNA were prepared by quantitative reconstitution of Sendai envelope proteins, fusion protein and hemagglutinin neuramindase in liposomal vesicles. The Sendai virosomes more efficiently transferred genes into cultured 293 transformed kidney cells than 1,2-dioleoyl-3 (trimethylammonium) propane-based cationic liposomes. At 200:1 weight ratio of envelope protein and lipid, the virosomes exhibited the best efficiency of gene transfection into the cells. The Sendai virosomes required relatively a short period of incubation time and much less cytotoxic, compared to the cationic liposome/DNA complex. The transfection efficiency of the Sendai virosomes containing DNA was maintained 70% after a month. This type of Sendai virosomes is relatively convenient for preparation and storage, compared to fusogenic liposomes prepared by liposome-virus fusion. First of all, because the constituents are quantitatively formulated, this type of virosome formulation can provide further consistent transfection for gene therapy. PMID- 11857034 TI - Resistance of prostate cancer cells to soluble TNF-related apoptosis-inducing ligand (TRAIL/Apo2L) can be overcome by doxorubicin or adenoviral delivery of full-length TRAIL. AB - Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL/Apo2L) has been shown to induce apoptosis in malignant cells without harming normal cells. To determine the antitumor potential of TRAIL against prostate cells, we undertook a comprehensive study that included eight prostate cancer cells lines (CWR22Rv1, Du145, DuPro, JCA-1, LNCaP, PC-3, PPC-1, and TsuPr1) and primary cultures of normal prostate epithelial cells (PrEC). Cells were tested for susceptibility to soluble TRAIL in the presence or absence of the chemotherapeutic agent doxorubicin. TRAIL was also delivered by an adenoviral vector. Our results reveal that Du145, DuPro, LNCap, TsuPr1, and PrEC were resistant to 100 ng/mL TRAIL. JCA 1 and PPC-1 were slightly sensitive (20% killing) and PC-3 and CWR22Rv1 exhibited the highest sensitivity to TRAIL (30% and 50% killing, respectively). The combination of 10 ng/mL TRAIL with doxorubicin resulted in 60-80% cytotoxicity in seven of eight prostate cancer cells. TRAIL-mediated apoptosis involved cleavage of Bid, caspase-3, and PARP, and required caspase-8 and -9 activity. Full-length TRAIL delivered by an adenoviral vector (AdTRAIL-IRES-GFP) killed prostate cancer cell lines and PrEC without requisite doxorubicin cotreatment. Therefore, expression of the transgene from a tissue-specific promotor would make gene therapy with AdTRAIL-IRES-GFP a possibility. PMID- 11857036 TI - Differential cytotoxicity and bystander effect of the rabbit cytochrome P450 4B1 enzyme gene by two different prodrugs: implications for pharmacogene therapy. AB - The time course of cytotoxicity induction and the bystander effect of the rabbit cytochrome P450 4B1 (cyp4B1)/4-ipomeanol (4-IM) or 2-aminoanthracene (2-AA) pharmacogene therapy systems were investigated and compared with the herpes simplex virus type 1 thymidine kinase/ganciclovir (HSV-tk/GCV) system. Experiments were performed in rat 9L gliosarcoma cells stably expressing cyp4B1 (9L-4B1), HSV-tk (9L-tk), or their egfp (enhanced green fluorescent protein) fusion genes. Cyp4B1-mediated activation of 2-AA showed a high cell killing efficiency within only 48 hours with an onset after already 15 minutes of prodrug exposure. Residual 9L-4B1 cells were mostly damaged sublethally upon 2-AA treatment showing an S phase arrest by cell cycle analysis. 4-IM treatment of 9L 4B1 cells generated an overall weaker cell killing, especially after prodrug exposure times of less than 48 hours. Residual cells surviving 4-IM treatment showed a G2/M arrest and restarted proliferation after prodrug treatment was stopped. HSV-tk/GCV pharmacogene therapy resulted in a slower cytotoxicity induction than cyp4B1/2-AA treatment with a significantly lower cell killing efficiency after 24 and 48 hours. HSV-tk/GCV-mediated cytotoxicity was widely similar to the cytotoxicity induced by cyp4B1/4-IM with the exception of a continuous 48-hour prodrug exposure where 4-IM treatment showed a significantly higher cell killing rate. Cells surviving HSV-tk/GCV suicide gene therapy were not viable and showed an S-phase arrest. Whereas HSV-tk/GCV induced a strong bystander effect, only moderate bystander cell death depending on cell-to-cell contact was demonstrated in 9L/9L-4B1 cocultures upon 2-AA treatment and was even absent with 4-IM, thereby contrasting with earlier reports. The absence of a strong bystander effect may limit, on one hand, the overall utility of the cyp4B1 systems for cancer gene therapy. On the other hand, the weak bystander effect together with the fast induction of cytotoxicity may provide marked advantages for the use of the cyp4B1 systems as biosafety enhancers for gene marking or replacement studies and donor lymphocyte infusions after allogeneic bone marrow transplantation. PMID- 11857037 TI - Transrectal gene therapy of the prostate in the canine model. AB - Direct transrectal delivery of therapeutic genes utilizing adenoviral vectors for advanced prostate cancer may offer effective treatment at the molecular level. Large animal models to assess feasibility and the intraprostatic and systemic dissemination patterns of these vectors have not been reported. For these studies, a replication-deficient (E1(-)/E3(-)) recombinant adenovirus (AdRSVlacZ) expressing bacterial beta-galactosidase (beta-gal) was delivered under transrectal ultrasound guidance. Two prostate biopsies, followed by concurrent injection of 4.8 x 10(9) pfu of the adenoviral vector divided into either 1 or 2 mL of diluent, were performed (n=4). Swabs of the rectum, sputum, and urine were collected and after 72 hours, the animals were sacrificed. Specimens were assayed for the presence of virus and beta-gal activity. Rectal swabs were transiently positive, whereas urine and sputum samples showed no detectable vector throughout the experiment. Beta-gal activity was observed at the prostate injection sites with detectable activity noted up to 7.5 mm away from the injection site. Systemic dissemination was observed regardless of the injected volume. In conclusion, transrectal prostate biopsy with concurrent prostate injection is a feasible method to deliver therapeutic adenoviral vectors for the treatment of prostate cancer; however, systemic distribution and temporary rectal shedding of virus should be anticipated. PMID- 11857038 TI - VEGF receptor antisense therapy inhibits angiogenesis and peritoneal dissemination of human gastric cancer in nude mice. AB - The efficacy of a phosphorothioate antisense oligonucleotide (ASO) for KDR/Flk-1 (KDR/Flk-1-ASO), an endothelial cell-specific vascular endothelial growth factor (VEGF) receptor, was investigated on the peritoneal dissemination and angiogenesis of a human gastric cancer cell line in nude mice. Green fluorescent protein (GFP)-transduced NUGC-4 (NUGC-4-GFP) human gastric cancer cells were implanted into the peritoneal cavity of nude mice. KDR/Flk-1-ASO, -SO, or phosphate-buffered saline was administrated from days 7 to 14, 200 microg/mouse, once a day. The mice were sacrificed on day 28. Disseminated peritoneal tumor nodules expressing GFP were visualized by fluorescence microscopy. KDR/Flk-1-ASO significantly decreased the extent of peritoneal dissemination of the tumors. The number of cells undergoing apoptosis was significantly increased in the KDR/Flk-1 ASO-treated tumors. Microvessel density was significantly reduced in the KDR/Flk 1-ASO-treated tumor nodules. The KDR/Flk-1 antisense strategy, therefore, decreases tumor dissemination apparently by inhibiting angiogenesis. PMID- 11857039 TI - Adenovirus-mediated CD40 ligand gene-engineered dendritic cells elicit enhanced CD8(+) cytotoxic T-cell activation and antitumor immunity. AB - CD40L, the ligand for CD40 on dendritic cells (DCs), plays an important role in their activation and is essential for induction of antigen-specific T-cell responses. In the present study, we investigated the efficacy of antitumor immunity induced by vaccination with DCs engineered to express CD40L and pulsed with Mut1 tumor peptide. Our data show that transfection of DCs with recombinant adenovirus AdV-CD40L resulted in activation of DCs with up-regulated expression of proinflammatory cytokines (IL-1beta and IL-12), chemokines (RANTES, IP-10, and MIP-1alpha), and immunologically important cell surface molecules (CD54, CD80, and CD86). Our data also demonstrate that DCs transfected with AdV-CD40L (DC(CD40L)) are able to stimulate enhanced allogeneic T-cell proliferation and Mut1-specific CD8(+) cytotoxic T-cell responses in vitro. Vaccination of mice with Mut1 peptide-pulsed control virus-transfected DC (DC(pLpA)) could only protect mice from challenge of a low dose (0.5 x 10(5) cells per mouse, 8/8 mice), but not a high dose (3 x 10(5) cells per mouse, 0/8 mice) of 3LL tumor cells. However, vaccination of Mut1 peptide-pulsed AdV-CD40L-transfected DC(CD40L) induced an augmented antitumor immunity in vivo by complete protection of mice (8/8) from challenge of both low and high doses of 3LL tumor cells. Thus, DCs engineered to express CD40L by adenovirus-mediated CD40 ligand gene transfer may offer a new strategy in production of DC cancer vaccines. PMID- 11857040 TI - Expression of the prodrug-activating enzyme DT-diaphorase via Ad5 delivery to human colon carcinoma cells in vitro. AB - Intratumoral injection of recombinant adenoviral type 5 (Ad5) vectors that carry prodrug-activating enzymes like DT-diaphorase (DTD) could be used to selectively target tumor cells for chemotherapy. To demonstrate the feasibility of this approach, Ad5 vectors were constructed, which express human DTD minigenes for both wild-type and mutant (C-to-T change in nucleotide 609 in DTD cDNA) DTD under the control of the cytomegalovirus (CMV) promoter. HT29 human colon carcinoma cells express wild-type DTD, whereas BE human colon carcinoma cells express mutant DTD, have low to undetectable DTD activity, and are 4- to 6-fold more resistant to mitomycin C (MMC) than HT29 cells. A test of the ability of Ad5 to infect these cells (using a beta-galactosidase CMV-driven minigene) indicated that 90-100% of BE cells were infected at a multiplicity of infection (MOI) of 100, whereas only 15-40% of HT29 cells were infected at this MOI. Infection of BE cells in vitro with recombinant Ad5 carrying a minigene for wild-type DTD at MOIs of 3-100 resulted in a progressive increase in DTD activity and a maximal 8-fold increase in sensitivity to MMC as measured by a colony-forming assay. HT29 cells were sensitized 2- to 3-fold following treatment with Ad5.DTD at an MOI of 100. These results indicate that adenovirus-mediated gene transfer and expression of wild-type DTD can sensitize resistant tumor cells to MMC and that this therapeutic strategy may exert a significant bystander effect. PMID- 11857041 TI - The use of armspan measurement to assess the nutritional status of adults in four Ethiopian ethnic groups. AB - OBJECTIVES: To investigate the use of armspan as a proxy for height in the assessment of nutritional status using body mass index (BMI) for four ethnic groups in Ethiopia. DESIGN: Cross-sectional study. SETTING: Four regions in Ethiopia, namely Oromia, Amhara, Tigre and Somali Region. SUBJECTS: A total of 1706 (884 males and 822 females) Ethiopians aged 18-50 y from four different ethnic groups. METHODS: Anthropometric measurements (weight, height and armspan) were obtained using standard techniques. BMI using height (BMI-ht) and using armspan (BMI-as) were calculated, t-tests were used to compare means, and linear regression to investigate the relationship between BMI-ht and BMI-as. RESULTS: Ethnic and sex differences in the relationship between height and armspan, and their derived variables (BMI-ht and BMI-as), were found. Armspan and height (r=0.83-0.9), and BMI-ht and BMI-as (r=0.89-0.97), were highly correlated in all ethnic groups. BMI-as cut-offs equivalent to the conventional BMI-ht classification of chronic energy deficiency were similar in the Oromo, Amhara and Tigre, but substantially higher in the Somalis. CONCLUSION: Armspan can be used as a proxy for height to estimate BMI, but the relationship between the two measures varies considerably with ethnicity and sex. Unless sex- and ethnicity specific cut-offs are applied, the use of BMI-as using conventional cut-offs will overestimate the prevalence of underweight in these populations. SPONSORSHIP: HelpAge International. PMID- 11857042 TI - Are lifestyle factors good predictors of retinol and vitamin C deficiency in apparently healthy adults? AB - OBJECTIVE: To examine interrelationships between (1) dietary habits, (2) socioeconomic and (3) environmental factors, and their impact on plasma retinol and plasma ascorbic acid. DESIGN: Cross-sectional study on adults from Western India. SETTING: Rural, semi urban, urban higher/middle/lower socioeconomic regions (HSE/MSE/LSE) having diverse dietary habits and environmental conditions. SUBJECTS: A total of 214 men and 108 women (20-50 y), apparently healthy and non anemic. MAIN OUTCOME MEASURES: Food intake by food frequency questionnaire, weight, height, age, smoking, environmental score, education, income, plasma retinol and plasma ascorbic acid. RESULTS: Mean plasma retinol in women (24.84+/ 5.1 microg/dl) and men (24.75+/-4.53 microg/dl) were not significantly different and 21% had plasma retinol below 20 microg/dl. Mean plasma ascorbic acid in women (0.35+/-0.12 mg/dl) and men (0.30+/-0.12 mg/dl) was similar with 75% having plasma ascorbic acid below 0.4 mg/dl. Vitamin A intake (as retinol equivalent) and plasma retinol showed a significant dose response (P<0.05) but not vitamin C intake and plasma ascorbic acid. Plasma retinol showed significant correlation with income (rho=0.24), education (rho=0.27), and environment (rho=0.21; rho=0.0001). Similar correlations with plasma ascorbic acid were 0.29, 031, -0.23 respectively (P=0.0001). Logistic regression showed education, environment, green leafy vegetables (GLV) and milk intake as predictors of plasma retinol deficiency, while non-sweet fruit intake, education and passive smoking for plasma ascorbic acid deficiency (P<0.05). CONCLUSIONS: Subnormal status of retinol and vitamin C emphasizes the need to increase consumption of fruit, GLV and milk products, and also better education and environment. Avoiding passive smoking demands attention in order to improve levels of these vitamins. SPONSORSHIP: Department of Science and Technology, India (project no. SP/SO/B39/94). PMID- 11857044 TI - Effect of ingestion of virgin olive oil on human low-density lipoprotein composition. AB - OBJECTIVE: To measure the incorporation of oleic acid and antioxidants (phenols and vitamin E) to low density lipoprotein (LDL) after acute and short-term ingestion of virgin olive oil. To study whether this incorporation contributes to an increase in LDL resistance to oxidation. SETTING: Department of Food and Nutrition, University of Barcelona, Spain and Department of Lipids and Cardiovascular Epidemiology, IMIM, Barcelona, Spain. SUBJECTS: Sixteen healthy volunteers aged 25-65 y. DESIGN AND INTERVENTIONS: To observe the change in the fatty acid profile, vitamin E, phenolic compounds and LDL oxidation-related variables after the postprandial phase and after daily ingestion of olive oil for one week. RESULTS: Few changes were observed in the postprandial phase. However, after a week of olive oil consumption there was an increase in oleic acid (P=0.015), vitamin E (P=0.047), phenolics (P=0.021) and lag time (P=0.000), and a decrease in the maximum amount of dienes (P=0.045) and oxidation rate (P=0.05). CONCLUSION: After ingestion of virgin olive oil, an increase in antioxidants and oleic acid in LDL was observed as well as an improvement of LDL resistance to oxidation. Our results support the idea that daily ingestion of virgin olive oil could protect LDL from oxidation. SPONSORSHIP: This study was supported by a research grant from Spain (ALI 97-1607-C02-02). PMID- 11857043 TI - Assessment of habitual energy and macronutrient intake in adults: comparison of a seven day food record with a dietary history interview. AB - OBJECTIVE: To examine the quantitative agreement between a 7 day food record and a diet history interview when these are conducted under the same conditions and to evaluate whether the two methods assess habitual diet intake differently among subgroups of age and body mass index (BMI). DESIGN: Cross-sectional study. SETTING: Population study, Denmark. SUBJECTS: A total of 175 men and 173 women aged 30-60 y, selected randomly from a larger population sample of Danish adults. INTERVENTIONS: All subjects had habitual diet intake assessed by a diet history interview and completed a 7 day food record within 3 weeks following the interview. The diet history interview and coding of records were performed by the same trained dietician. MAIN OUTCOME MEASURE: Median between-method difference in assessment of total energy intake, absolute intake of macronutrients, and nutrient energy percentages. Difference between reported energy intake from both methods and estimated energy expenditure in different subgroups. RESULTS: Energy and macronutrient intake was assessed slightly higher by the 7 day food record than by the diet history interview, but in absolute terms the differences were negligible. The between-method difference in assessment of total energy intake appeared to be stable over the range of age and BMI in both sexes. As compared to estimated total energy expenditure, both diet assessment methods underestimated energy intake by approximately 20%. For both methods the under-reporting increased by BMI in both sexes and by age in men. CONCLUSIONS: Energy and macronutrient intake data collected under even conditions by either a 7 day food record or a diet history interview may be collapsed and analysed independent of the underlying diet method. Both diet methods, however, appear to underestimate energy intake dependent on age and BMI. SPONSORSHIP: Danish Medical Research Council, the FREJA programme. PMID- 11857045 TI - Effects of intravenous supplementation with alpha-tocopherol in patients receiving total parenteral nutrition containing medium- and long-chain triglycerides. AB - OBJECTIVE: To compare the effects of a lipid emulsion containing medium-chain triglycerides (MCT) and supplemented with alpha-tocopherol to a conventional long chain triglyceride (LCT) emulsion. DESIGN: Randomised double blind study. SETTING: Department of Internal Medicine, Antwerp University Hospital. SUBJECTS AND INTERVENTIONS: Twenty-four patients with an indication for total parenteral nutrition for a minimum of 10 days were randomly assigned to two groups: group E received as lipid source MCT/LCT (50/50) suplemented with 100 mg DL-alpha tocopherol/day and group C received LCT. Blood samples were analysed at inclusion, after 4-6 and after 9-11 days. RESULTS: In group E, serum alpha tocopherol doubled from 11.4+/-6.9 at inclusion to 20.9+/-7.9 and to 23.8+/-8.8 microg/ml after 4 and 9 days, respectively, but did not change in group C (P=0.008). Production of thiobarbituric acid-reacting substances (TBARS) after 120 min incubation with copper decreased from 66+/-34 at inclusion to 29+/-25 nmol MDA/mg LDL and VLDL-cholesterol after 4 and to 42+/-17 after 9 days (P=0.022 when compared to group C, which underwent no significant changes). Velocity of production of fluorescent products decreased in group E but not in group C (P=0.026). CONCLUSIONS: Supplementation of TPN containing MCT/LCT with 100 mg DL alpha-tocopherol/day leads to a doubling in serum alpha-tocopherol and to a decrease in the susceptibility of LDL and VLDL to peroxidation in vitro. SPONSORSHIP: This study was partly financed by B Braun Medical NVSA, Diegem, Belgium. PMID- 11857046 TI - The effect of graded levels of exercise on energy intake and balance in free living men, consuming their normal diet. AB - OBJECTIVE: To assess the effect of graded increases in exercised-induced energy expenditure (EE) on appetite, energy intake (EI), total daily EE and body weight in men living in their normal environment and consuming their usual diets. DESIGN: Within-subject, repeated measures design. Six men (mean (s.d.) age 31.0 (5.0) y; weight 75.1 (15.96) kg; height 1.79 (0.10) m; body mass index (BMI) 23.3(2.4) kg/m(2)), were each studied three times during a 9 day protocol, corresponding to prescriptions of no exercise, (control) (Nex; 0 MJ/day), medium exercise level (Mex; approximately 1.6 MJ/day) and high exercise level (Hex; approximately 3.2 MJ/day). On days 1-2 subjects were given a medium fat (MF) maintenance diet (1.6 x resting metabolic rate (RMR)). MEASUREMENTS: On days 3-9 subjects self-recorded dietary intake using a food diary and self-weighed intake. EE was assessed by continual heart rate monitoring, using the modified FLEX method. Subjects' HR (heart rate) was individually calibrated against submaximal VO(2) during incremental exercise tests at the beginning and end of each 9 day study period. Respiratory exchange was measured by indirect calorimetry. Subjects completed hourly hunger ratings during waking hours to record subjective sensations of hunger and appetite. Body weight was measured daily. RESULTS: EE amounted to 11.7, 12.9 and 16.8 MJ/day (F(2,10)=48.26; P<0.001 (s.e.d=0.55)) on the Nex, Mex and Hex treatments, respectively. The corresponding values for EI were 11.6, 11.8 and 11.8 MJ/day (F(2,10)=0.10; P=0.910 (s.e.d.=0.10)), respectively. There were no treatment effects on hunger, appetite or body weight, but there was evidence of weight loss on the Hex treatment. CONCLUSION: Increasing EE did not lead to compensation of EI over 7 days. However, total daily EE tended to decrease over time on the two exercise treatments. Lean men appear able to tolerate a considerable negative energy balance, induced by exercise, over 7 days without invoking compensatory increases in EI. PMID- 11857047 TI - Dietary patterns among children aged 6-7 y in four Spanish cities with widely differing cardiovascular mortality. AB - OBJECTIVE: Classic cardiovascular risk factors, such as smoking, arterial hypertension and hypercholesterolaemia, cannot explain a substantial part of the geographic differences in cardiovascular mortality. Anthropometric and nutritional factors in early stages of life may contribute to adult cardiovascular disease. Therefore, this work examines certain anthropometric variables and diet among children aged 6-7 y, living in four Spanish cities with widely differing ischaemic heart disease (IHD) mortality. DESIGN AND SETTING: Cross-sectional anthropometric and dietary survey in four cities in Spain. SUBJECTS: A total of 1112 children (50.1% males, 49.9% females) attending public and private schools in Cadiz and Murcia, cities with a relatively high IHD mortality, and Madrid and Orense, cities with a relatively low IHD mortality. A standardized method was used to measure anthropometric variables, and a food frequency questionnaire completed by subjects' mothers, to measure diet. OUTCOME MEASURES: Body mass index (BMI), overweight (BMI>17.6 kg/m(2)), obesity (BMI>20.1 kg/m(2)) and intake of food and nutrients. RESULTS: Children in the four cities showed a high prevalence of overweight (range across cities, 28.9-34.5%) and obesity (8.5-15.7%). They also had a moderately hypercaloric diet (range, 2078 2218 kcal/day), marked by an excessive intake of lipids (45.0-47.3% kcal), particularly saturated fats (16.6-16.9% kcal), proteins (17.0-17.3% kcal), sugars (20.0-21.9% kcal) and cholesterol (161.6-182.9 mg/1000 kcal/day), and a low intake of complex carbohydrates (17.5-18.1% kcal) and fibre (19.6-19.9 g/day). Compared with children in the two low-IHD-mortality cities, those in the two high IHD-mortality cities had a greater BMI (mean difference, 0.61 kg/m(2); P=0.0001) and ponderal index (0.58 kg/m(3); P=0.0001) and a higher intake of energy (104 kcal/day; P=0.007), cholesterol (16.00 mg/1000 kcal/day; P=0.0001) and sodium (321 mg/day; P=0.0001). Inter-city differences in anthropometric variables remained after adjustment for birthweight. CONCLUSIONS: Intake of fats, especially saturated fats, and cholesterol should be reduced among Spanish children. It could contribute to a needed reduction of the high prevalence of overweight and obesity in children. If the differences in anthropometric variables and diet between children from the cities with high and low coronary mortality are maintained in future or continue into adulthood, this could contribute to consolidate or even increase the IHD mortality gradient across cities. The finding that differences in anthropometric variables are independent of birthweight suggests that the childhood, rather than intrauterine environment, is involved in the development of such differences. SPONSORSHIP: This study was partly funded by grants from the International Olive Oil Board (Consejo Oleicola Internacional), Comunidad Autonoma de Madrid, Fundacion Pedro Barrie de la Maza, and Fundacion Eugenio Rodriguez Pascual. PMID- 11857048 TI - The impact of the leucine 7 to proline 7 polymorphism of the neuropeptide Y gene on postprandial lipemia and on the response of serum total and lipoprotein lipids to a reduced fat diet. AB - OBJECTIVE: The aim of the study was to examine the impact of the leucine7 to proline7 (Leu7Pro) polymorphism of the NPY gene on postprandial (PP) lipemia, post-heparin plasma lipoprotein lipase (LPL) and hepatic lipase (HL) activities, and the response of serum lipids to a reduced fat diet. DESIGN AND SUBJECTS: Seven middle-aged obese subjects with Leu7Pro genotype were matched with seven subjects with Leu7Leu genotype for gender, age, apolipoprotein E phenotype and BMI. These 14 subjects participated in the oral 8 h fat tolerance test. Sixty eight slightly obese middle-aged subjects (10 with the Leu7Pro genotype) had participated in intervention studies and consumed a reduced fat diet for 8 weeks. RESULTS: There were no statistically significant differences in PP areas under the curve of plasma total triglycerides (TG), chylomicron TG, VLDL-TG or insulin between the genotype groups. The TG-to-cholesterol (C) ratio in VLDL was significantly lower in the subjects with Leu7Pro genotype compared to those with the Leu7Leu genotype at time points 30 min and 1 h in the fat tolerance test. Heparin-induced activities of LPL or HL or the response of serum total or LDL-C to the reduced fat diet did not differ between the groups. CONCLUSIONS: The NPY genotype neither affects the magnitude of postprandial lipemia induced by a fat tolerance test nor the response of serum total lipids or lipids in different lipoprotein classes to the reduced fat diet. However, this preliminary study suggests that there might be compositional differences in the lipoprotein particles between the genotype groups that affect postprandial lipid metabolism. SPONSORSHIP: The Council for Health Sciences of the Academy of Finland, Kuopio University Hospital and the National Technology Agency, Finland. PMID- 11857050 TI - Relations between vegetable, fruit and micronutrient intake. Implications for odds ratios in a case-control study. AB - OBJECTIVE: To investigate whether the protection observed for some micronutrients, such as beta-carotene, in several observational studies may simply reflect vegetable and fruit intake. DESIGN: A case-control study conducted in Italy. SUBJECTS: The subjects were 1225 colon cancer patients, 728 rectal cancer patients and 4154 hospital controls. RESULTS: For the 16 micronutrients considered, the more closely a micronutrient was correlated with total vegetable and fruit intake, the more it appeared protective against colorectal cancer. CONCLUSION: When studying the effect of a nutrient on disease risk in an observational setting, its relation to other nutrients and foods must be taken into account. PMID- 11857049 TI - The effect of flaxseed supplementation in processed foods on serum fatty acids and enterolactone. AB - OBJECTIVE: The purpose was to study the effects of flaxseed supplementation as a part of daily diet on serum lipids, fatty acids and plasma enterolactone. DESIGN: Eighty volunteers participated in this clinical nutrition study which was carried out in a controlled, double-blind and cross-over manner. The subjects were randomized to diet sequences AB or BA. Diet A meals contained 1.3 g/100 g ground flaxseed and 5 g/100 g flaxseed oil. Also 3-4 g/100 of inulin and wheat fiber was added. AB diet with non-supplemented foods served as control. Test subjects were on both diets for 4 weeks separated by a 4-week wash-out period. Fifteen test subjects continued an open part of the study for 4 additional months. INTERVENTIONS: The dietary intake, basic blood values, serum lipids, fatty acids and enterolactone were measured at baseline, after both intervention periods and during the open study, at baseline and after 2 and 4 months. Serum thiocyanate and blood cadmium were controlled after both intervention periods. RESULTS: The percentage of flaxseed supplemented test food out of total dietary intake was 20% of energy. The test food contained significantly higher amounts of fiber, polyunsaturated fatty acids (PUFAs) and especially alpha-linolenic acid than the control food. No significant changes were observed in the basic laboratory values or in blood lipids. There was a significant increase in serum alpha-linolenic acid, eicosapentaenoic acid and docosapentaenoic acid. Serum enterolactone concentration was doubled during flaxseed supplementation. Serum thiocyanate and blood cadmium values did not exceed reference values and there was no difference between the diets. CONCLUSIONS: In this study we were able to show that, by adding ground flaxseed and flaxseed oil to one or two daily meals, it is possible to obtain significant effects on serum levels of enterolactone and alpha linolenic acid. SPONSORSHIP: The study was sponsored by the National Technology Agency of Finland (Tekes). PMID- 11857051 TI - Italian cross-sectional growth charts for height, weight and BMI (6-20 y). AB - OBJECTIVE: To trace growth charts for height, weight and body mass index (BMI) that apply to the whole Italian population. Different charts were drawn for central-north and south Italy since children in central-north regions are known to be taller and leaner. DESIGN: Cross-sectional study. SETTING: A sample of schoolchildren covering 16 of the 20 Italian regions, with data collected between 1994 and 2000. SUBJECTS: A total of 27 421 girls and 27 374 boys, aged 6-20 y. METHODS: Height and weight were measured using portable Harpenden stadiometers and properly calibrated scales, respectively. SIEDP references are presented both as centiles and as LMS curves for the calculation of standard deviation scores. According to International Obesity Task Force, SIEDP charts for BMI include the limits for overweight and obesity, ie the centiles having, at 18 y of age, the value of 25 and 30 kg/m(2), respectively. RESULTS: The comparison between SIEDP and Tanner et al's charts for height, still in use among most Italian paediatricians, shows that before puberty Italian children are 2-4 cm taller than their English peers. Because of these differences, Tanner's charts fail to detect, when applied to Italian children, 50-90% of short children aged 6-11 y, ie with stature below the 3rd centile of their reference population. Rolland Cachera et al's centiles for BMI are lower than those of SIEDP standards, mainly during adolescence (up to 6.6 kg/m(2) for the 97th centile), and apply poorly to Italian children. The prevalence of overweight is 27 (boys) and 19% (girls) in south Italy vs 17 (boys) and 10% (girls) in central-north Italy. CONCLUSIONS: These references intend to supply Italian paediatricians with a tool that avoids the use of outdated or inadequate charts, and thus should be suitable for monitoring their patients' growth. SPONSORSHIP: Italian Society for Pediatric Endocrinology and Diabetes (SIEDP). PMID- 11857052 TI - Description of three new polymorphisms in the intronic and 3'UTR regions of the human interferon gamma gene. AB - Interferon-gamma (IFN-gamma) is a key regulator of the development and functions of the immune system. In particular, this cytokine plays a major role in immune defense against infections by various human pathogens and polymorphisms in the IFN-gamma gene, including the transcription regulatory region, and might affect host resistance to infectious agents such as schistosomes. In this study on the genetics of human schistosomiasis we uncovered three new single nucleotide polymorphisms in the IFN-gamma genes. Two polymorphisms are located in the third intron and the third is in the 3'UTR region of this gene: an A to G transition at position +2109 from the transcription start and two G to A transitions at positions +3810 and +5134. In a SUDANESE population living in an endemic area of malaria and schistosomiasis, the allelic frequenciesare: 0.85 (+2109A), 0.15 (+2109G), 0.92 (+3810G), 0.08 (+3810A), (+5134G) and 0.04 (+5134A). PMID- 11857053 TI - Genetic susceptibility to fibrocalculous pancreatic diabetes in Bangladeshi subjects: a family study. AB - Fibrocalculous pancreatic diabetes (FCPD) is an uncommon cause of diabetes, seen mainly in developing countries. A family-based study was carried out in 67 Bangladeshi families, consisting of a proband with FCPD and both parents, to determine whether an association exists between FCPD susceptibility and either the major histocompatiblity complex (MHC) or insulin gene (INS) loci. HLA-DQB1 typing was done using allele-specific primers, and INS was typed using the restriction enzyme HphI. Three microsatellites (TNFa, TNFc and TNFd), from within and flanking the TNF-LT locus, were used for MHC Class IV typing and a PCR-RFLP assay was used to define the -308G/A TNF promoter polymorphism. The extended transmission disequilibrium test (ETDT) was used for statistical analysis. An overall association was observed between FCPD and HLA-DQB1 (P = 0.003), that was largely due to a positive association with HLA-DQB1*0302 and a negative association with HLA-DQB1*0202. Although no association was found between FCPD and TNF-LT microsatellite markers a trend was observed for TNFc (P = 0.037, Pc = 0.15). No association was found between FCPD and INS (P = 0.26). This study confirms an association between FCPD and the MHC using a family-based study design and the stringent ETDT analysis; a novel protective association was found with HLA-DQB1*0202 in Bangladeshi FCPD subjects. The genetic susceptibility to FCPD has features both similar and dissimilar to T1DM. PMID- 11857054 TI - A genome-wide linkage analysis of orchard grass-sensitive childhood seasonal allergic rhinitis in Japanese families. AB - Seasonal allergic rhinitis (SAR) is an inflammatory disease of the nose and eyes that follows sensitization to air-born pollens. We conducted a genome-wide linkage screening of 48 Japanese families (188 members) with orchard grass (OG) sensitive SAR children (67 affected sib-pairs) in a farming community in central Japan where OG was planted for apple farming and OG pollen is a major cause of SAR. We used the GENEHUNTER program to performed nonparametric multipoint linkage analysis for OG-sensitive SAR as a qualitative trait and for log total serum IgE levels and OG-RAST IgE levels as quantitative traits. Genotyping data of 400 microsatellite markers suggested linkage of SAR to chromosomes 1p36.2, 4q13.3, and 9q34.3 (P < 0.001), linkage of serum total IgE levels to 3p24.1, 5q33.1, 12p13.1, and 12q24.2 (P < 0.001), and linkage of OG-RAST IgE levels to 4p16.1, 11q14.3, and 16p12.3 (P < 0.001). Weak evidence for linkage of SAR to 5q33.1 was also observed (P = 0.01). All these regions, with the exception of 9q34.3, have been previously reported to be linked to asthma and/or atopy. These data suggest that, although loci linked to SAR are likely to be common to asthma, a strong contribution by specific gene(s) to OG-sensitive SAR is unlikely. PMID- 11857055 TI - Association between baseline levels of C-reactive protein (CRP) and a dinucleotide repeat polymorphism in the intron of the CRP gene. AB - Elevation of baseline C-reactive protein (CRP) is associated with increased risk of cardiac disease. This increase might reflect low-grade inflammation, but differences in CRP serum levels might also have a genetic component. To test this possibility, we investigated whether a polymorphic GT-repeat in the intron of the CRP gene contributes to variation in baseline CRP. We found that the polymorphism was associated with differences in baseline CRP in both normal individuals and in patients with the inflammatory disease systemic lupus erythematosus, viz. donors carrying two GT(16) alleles, two GT(21)alleles, or GT(16/21) heterozygotes had two-fold lower serum CRP than those with other genotypes. The frequency of GT(16) and GT(21) was two-fold higher in Caucasians than in African-Americans, but there was no difference in allele distribution between patients and controls. It is not yet known how this genetic polymorphism mediates its effect on CRP expression, and it probably is not a systemic lupus erythematosus susceptibility factor. Rather, the CRP intron polymorphism likely modifies the disease phenotype. On the other hand, the fact that baseline CRP does have a genetic component suggests that in coronary disease, stratification of risk assessment based on CRP levels might be enhanced by consideration of this polymorphism. PMID- 11857056 TI - Association between the MHC class I gene HFE polymorphisms and longevity: a study in Sicilian population. AB - Classes I and II human leukocyte antigens (HLA) genes encode highly polymorphic heterodimeric glycoproteins involved in the control of immune responses. The HLA class I gene HFE seemingly no longer participates in immunity because it has lost its ability to bind peptides and it has acquired the ability to form complex with the receptor for iron-binding transferrin by regulating iron uptake by intestinal cells. Thus, it indirectly regulates immune responses too, because iron availability plays a role in specific and non-specific immune responses. The distribution of HFE polymorphisms in Sicilian centenarians and nonagenarians was studied to evaluate if HFE alleles might be represented differently in people selected for longevity. DNA samples were obtained from 106 young controls (age range from 22 to 55 years; 40 men and 66 women) and 35 elderly subjects (age range from 91 to 105 years; seven men and 28 women). Samples were typed for C282Y, H63D and S65C alleles using polymerase chain reaction and sequence specific primers. Among the young individuals, none was heterozygous for the C282Y or for S65C mutation. Twenty-six were heterozygous for H63D mutation. Among the elderly subjects, 11 were heterozygous for the C282Y mutation or for H63D mutation. None was heterozygous for the S65C mutation. No compound heterozygous individuals (C282Y/H63D) were found. A highly significant difference was observed in frequencies of C282Y alleles between the young and the elderly subjects on the whole. By analysing polymorphisms according to gender, heterozygous subjects for C282Y were found both in old men and in old women, but by comparing the allele frequencies to those of young people significance was attained only in women. Concerning H63D polymorphisms, no significant differences were observed, between old and young people, both in men and in women. Possession of C282Y allele, known to be associated with an increase of iron uptake, significantly increases women possibility to reach longevity. Thus, present data adds another piece of evidence to the complex puzzle of genetic and environmental factors involved in control of lifespan expectancy in humans. PMID- 11857057 TI - Polymorphisms in or near tumour necrosis factor (TNF)-gene do not determine levels of endotoxin-induced TNF production. AB - Innate differences in tumour necrosis factor (TNF) production have been associated with susceptibility for and outcome of inflammatory diseases. Several studies have tried to identify whether polymorphisms in or near the TNF gene or other markers on the short arm of chromosome 6 (6p21) are related to differences in TNF production. Data on these associations are conflicting. Therefore, we conducted a study among 129 healthy individuals in which TNF production was determined upon stimulation with endotoxin in whole blood cultures. TNFa microsatellite, TNF single nucleotide polymorphisms at position +489, -238, -308 and -376 typing was performed. The data revealed that alleles of TNFa microsatellite and carriership of TNF polymorphisms were not related to TNF production. We conclude that the genes determing the differences in endotoxin induced TNF production have not been yet identified. PMID- 11857058 TI - Gender-specific association between -1082 IL-10 promoter polymorphism and longevity. AB - Ageing is characterized by a pro-inflammatory status, which could contribute to the onset of major age-related diseases. Thus, genetic variations in pro- or anti inflammatory cytokines might influence successful ageing and longevity. IL-10 is an appropriate candidate because it exerts powerful inhibitory effects on pro inflammatory function. IL-10 production is controlled by several polymorphic elements in the 5' flanking region of IL-10 gene on 1q32 locus, involving alleles at two microsatellite regions and several polymorphisms in promoter region. We analysed in 190 Italian centenarians (>99 years old, 159 women and 31 men) and in 260 <60 years old control subjects (99 women and 161 men), matched for geographical distribution, genotype frequencies for -1082G-->A, -819C-->T and 592C-->A IL-10 proximal promoter gene biallelic polymorphisms by sequence specific probes. -1082G homozygous genotype was increased in centenarian men (P < 0.025) but not in centenarian women. No difference was found between centenarians and control subjects regarding the other two polymorphisms. The presence of 1082GG genotype, suggested to be associated with high IL-10 production, significantly increases the possibility to reach the extreme limit of human lifespan in men. Together with previous data on other polymorphic loci (Tyrosine Hydroxylase, mitochondrial DNA, IL-6, haemochromatosis, IFN-gamma), this finding points out that that gender is a major variable in the genetics of longevity, suggesting that men and women follow different strategies to reach longevity. Concerning the biological significance of this association, we have not searched for functional proves that IL-10 is involved. Thus, we should conclude that our data only suggest that a marker on 1q32 genomic region may be involved in successful ageing in man. However, recent data on IL-6 and IFN-gamma genes suggest that longevity is negatively associated with genotypes coding for a pro inflammatory profile. Thus, it is intriguing that the possession of -1082G genotype, suggested to be associated with IL-10 high production, is significantly increased in centenarians. PMID- 11857059 TI - Single nucleotide polymorphisms in MHC2TA, the gene encoding the MHC class II transactivator (CIITA). AB - The MHC class II transactivator (CIITA) is the master regulator for HLA-D (DP, DQ, DR) gene expression. In this report the coding and promoter regions of the CIITA gene, MHC2TA, were evaluated for polymorphisms in 50 normal Caucasian individuals. Allele frequencies were obtained for four separate single nucleotide (nt) polymorphisms (SNPs) identified in the MHC2TA coding region: nt 1614 (C- >G), nt 2509 (G-->A), nt 2536 (T-->G), and nt 2791 (G-->A). MHC2TA sequence analysis of 100 chromosomes from these 50 individuals revealed a SNP in MHC2TA promoter (p) III at nt (-)155 (A-->G), but none in CIITA pI or pIV. In addition, we demonstrate the presence of splice variant at a previously undiscovered intron, accounting for a three nt (TAG) insertion at position 474 that was originally described in association with one of the disease-causing CIITA cDNA mutations in bare lymphocyte syndrome. PMID- 11857060 TI - T helper 1-type immunity to trophoblast antigens in women with a history of recurrent pregnancy loss is associated with polymorphism of the IL1B promoter region. AB - Recurrent pregnancy loss (RPL) is a common disorder during early gestation. Recent evidence suggests that T helper 1 (Th1)-type immunity is associated with unsuccessful pregnancy especially in women with RPL of otherwise unknown etiology, while Th2-type immunity is associated with pregnancy success. Interleukin (IL)-1 may influence Th1/Th2 immune responsiveness and has been implicated in the establishment of successful pregnancy. In the present study, we investigated polymorphism of the IL-1beta gene (IL1B) in women with a history of RPL. Significant increases in the frequencies of IL1B promoter region variants IL1-511C and IL1B-31T were found in women with a history of RPL. Increased frequencies of these two variants and their homozygotes were found only in cases having evidence of Th1 immunity to trophoblast as determined by IFN-gamma production of peripheral blood mononuclear cells (PBMCs) stimulated with a trophoblast cell-line extract. Significantly higher IFN-gamma production by PBMCs in response to trophoblast correlated with variant IL1B-511C and its homozygocity in women with RPL. These results suggest that variants -511C and -31T in the IL1B promoter region confer risk for RPL associated with Th1 immunity to trophoblast antigens. PMID- 11857061 TI - Enhanced cytokine mRNA levels in attack-free patients with familial Mediterranean fever. AB - Familial Mediterranean fever (FMF) is a recessively inherited inflammatory disorder, characterized by recurrent attacks of fever and serositis. Screening of mutations in the causing gene (MEFV) now allows accurate diagnosis of FMF among other inflammatory conditions. It is well documented that secreted levels of some pro-inflammatory cytokines are elevated in FMF. Here, we investigated cytokine expression at the transcriptional level, in patients that could be genetically ascertained. We have measured the transcript abundance of tumor necrosis factor alpha, interleukin-1beta, interleukin-6 and interleukin-8, in circulating leukocytes and shown that these were more elevated in attack-free FMF patients than in controls (P=0.01, P=0.008, P=0.02, P=0.001 respectively). There was no significant difference according to MEFV genotype or colchicine treatment. Our results suggest that cytokine transcriptional pathways are misregulated in attack free FMF patients, and further supports the hypothesis that these patients have subclinical inflammation between attacks. PMID- 11857062 TI - CDS1 and promoter single nucleotide polymorphisms of the CTLA-4 gene in human myasthenia gravis. AB - The cytotoxic T lymphocyte associated protein 4 (CTLA-4) gene (Ctla-4) is a candidate gene for autoimmune disease. We here report results of two single nucleotide polymorphisms (SNPs) in the Ctla-4, a +49 A/G SNP in CDS1 and a C/T promoter SNP at position -318. There were no differences in these two SNPs between patients and healthy individuals. The frequency of allele G and genotype G/G at position +49 in CDS1 was increased in patients with thymoma when compared with patients with normal and hyperplastic thymic histopathology. Patients with the G/G genotype had signs of immune activation manifested as higher levels of serum IL-1beta and higher percentage of CD28(+) T lymphocytes. There was a strong linkage between the 86bp allele in the 3'-UTR and the A(+49) allele in CDS1. Our results suggest that the SNP at position +49 in CDS1 might be associated with the manifestations of MG. PMID- 11857063 TI - Pemphigus is not associated with allotypic markers of immunoglobulin kappa. AB - The kappa light chain constant region of immunoglobulins bears polymorphic markers involved in susceptibility to various autoimmune diseases. To determine whether it also contributes to the occurrence of pemphigus, a group of autoimmune blistering skin diseases owing to pathogenic autoantibodies, the genotypic frequencies of Km allotypes were evaluated in patients with pemphigus foliaceus or pemphigus vulgaris and ethnically-matched healthy controls in both Tunisia and France. No difference in the distribution of Km genotype or allele frequencies was observed between patients and controls in either countries. Therefore, Km allotypes do not appear to constitute a genetic factor contributing to pemphigus. PMID- 11857064 TI - A null mutation within the ciliary neurotrophic factor (CNTF)-gene: implications for susceptibility and disease severity in patients with multiple sclerosis. AB - Multiple sclerosis (MS) is an inflammatory demyelinating disease of the central nervous system. Impaired remyelination and axonal degeneration may account for progressive disability in MS patients. As ciliary neurotrophic factor (CNTF) takes part in myelogenesis, we examined the frequency of a CNTF-null mutation in 349 MS patients with respect to their clinical presentation and in comparison with 434 healthy controls. Similar genotype frequencies for the CNTF mutation were obtained in MS patients (genotype 0101=74.8%, 0102=22.3%, 0202=2.9%) and controls (genotype 0101=71.7%, 0102=26.5%, 0202=1.8%) even after stratification for the HLA-DRB1*15 allele. In addition, there was no significant correlation of CNTF genotypes to age at onset, course or severity of the disease. We therefore conclude, that the requirement for CNTF in myelogenesis or cell survival may be bypassed by a second ligand or redundancy of functional activity of other neurotrophic factors. PMID- 11857065 TI - Rheumatoid arthritis association in Colombian population is restricted to HLA DRB1*04 QRRAA alleles. AB - In most ethnic groups genetic susceptibility to rheumatoid arthritis (RA) is associated with certain HLA-DRB1 alleles encoding a similar sequence motif called the 'shared epitope' (SE) spanning amino acid positions 70 to 74 in the third diversity region of the outermost domain of the HLA-DRB1 molecule. We examined the association of the SE and RA in 83 Colombian women with established RA and 90 healthy controls. The group HLA-DRB1*04 was associated with RA with respect to controls (47% vs 18%, respectively. OR: 4.1, 95%CI: 2.1-8.2, P < 0.001). HLA-DRB1 alleles carrying the SE QRRAA, but not those carrying QKRAA or RRRAA, were associated with disease (OR: 3.7, 95%CI: 1.73-7.83, P = 0.0009). This association was stronger among HLA-DRB1*04 carriers (OR: 23, 95%CI: 1.3-414, P = 0.002). In our population, the SE QRRAA expressed in DRB1*04 alleles appears critical in identifying women with increased susceptibility to RA. PMID- 11857066 TI - Immunization strategy against cervical cancer involving an alphavirus vector expressing high levels of a stable fusion protein of human papillomavirus 16 E6 and E7. AB - We are developing immunization strategies against cervical carcinoma and premalignant disease, based on the use of recombinant Semliki Forest virus (SFV) encoding the oncoproteins E6 and E7 from high-risk human papilloma viruses (HPV). Thus far, protein-based, as well as genetic immunization studies have demonstrated low to moderate cellular immune responses against E6 and E7. To improve these responses, we modified the structure and expression level of the E6 and E7 proteins produced by the SFV vector. Specifically, a construct was generated encoding a fusion protein of E6 and E7, while furthermore a translational enhancer was included (enhE6,7). Infection of cells with recombinant SFV-enhE6,7 resulted in the production of large amounts of the E6,7 fusion protein. The fusion protein was more stable than either one of the separate proteins. Immunization of mice with SFV-enhE6,7 resulted in strong, long lasting HPV-specific cytotoxic T lymphocyte responses. Tumor challenge experiments in mice demonstrated that immunization with SFV-enhE6,7 resulted in prevention of tumor outgrowth and subsequent protection against tumor re challenge. PMID- 11857067 TI - Recombinant Semliki Forest virus as a vector system for fast and selective in vivo gene delivery into balloon-injured rat aorta. AB - Previously, we demonstrated that recombinant Semliki Forest virus (SFV) vector rapidly and selectively transfers genes into cultured vascular smooth muscle cells (VSMC), leaving endothelial cells (EC) unaffected. From this, we hypothesized that recombinant SFV in vivo only transfers genes into the media of balloon-injured but not intact vessel, that gene expression in VSMC is fast, and that the specificity of SFV for VSMC is caused by specific binding sites. To address these hypotheses, we studied the time course of in vivo SFV-LacZ and Ad LacZ expression in balloon-injured rat aorta. In addition, the fusion characteristics of fluorescent pyrene-labeled SFV were explored in cultured VSMC and EC. In intact aorta, no LacZ expression was found in the intima or media at 24 h. In contrast, in denuded aorta, LacZ expression was detected in as early as 12 h after incubation. LacZ expression was predominantly present in the media. Ad LacZ expression started after 12 h, but was predominantly present in the adventitia. Ad-LacZ expression in the media started after 72 h. In vitro transfection with SFV showed that fusion was higher and, moreover, saturable in VSMC as compared with EC, indicating the presence of specific SFV binding sites on VSMC, but not EC. From this we conclude that in vivo selectivity of SFV in balloon-injured vessels is based on the removal of the endothelium, which results in accessibility of VSMC in the media that carry specific binding sites for the SFV vector. PMID- 11857068 TI - Magnetofection: enhancing and targeting gene delivery by magnetic force in vitro and in vivo. AB - Low efficiencies of nonviral gene vectors, the receptor-dependent host tropism of adenoviral or low titers of retroviral vectors limit their utility in gene therapy. To overcome these deficiencies, we associated gene vectors with superparamagnetic nanoparticles and targeted gene delivery by application of a magnetic field. This potentiated the efficacy of any vector up to several hundred fold, allowed reduction of the duration of gene delivery to minutes, extended the host tropism of adenoviral vectors to nonpermissive cells and compensated for low retroviral titer. More importantly, the high transduction efficiency observed in vitro was reproduced in vivo with magnetic field-guided local transfection in the gastrointestinal tract and in blood vessels. Magnetofection provides a novel tool for high throughput gene screening in vitro and can help to overcome fundamental limitations to gene therapy in vivo. PMID- 11857069 TI - Adenovirus-mediated overexpression of extracellular superoxide dismutase improves endothelial dysfunction in a rat model of hypertension. AB - Gene transfer may be appropriate for therapeutic protocols targeted at the vascular endothelium. Endothelial dysfunction is the principal phenotype associated with atherosclerosis and hypertension. Oxidative stress has been implicated in the development of endothelial dysfunction. We have explored the ability of overexpressing anti-oxidant genes (superoxide dismutases; SODs) in vitro and in vivo to assess their potential for reversing endothelial dysfunction in a rat model, the stroke-prone spontaneously hypertensive rat (SHRSP). Western blotting and immunofluorescence assays in vitro showed efficient overexpression of MnSOD and ECSOD with respect to localisation to the mitochondria and extracellular surface, respectively. Transgene functional activity was quantified with SOD activity assays. MnSOD and ECSOD overexpression in intact SHRSP vessels in vivo led to endothelial and adventitial overexpression. Pharmacological assessment of transduced vessels following in vivo delivery by basal NO availability quantification demonstrated that the "null" adenovirus and MnSOD adenovirus did not significantly increase NO availability. However, AdECSOD treated carotid arteries showed a significant increase in NO availability (1.91 +/- 0.04 versus 0.75 +/- 0.08 g/g, n = 6, P = 0.029). In summary, efficient overexpression of ECSOD, but not MnSOD in vivo, results in improved endothelial function in a rat model of hypertension and has important implications for the development of endothelial-based vascular gene therapy. PMID- 11857071 TI - Cirrhotic rat livers with extensive fibrosis can be safely transduced with clinical-grade adenoviral vectors. Evidence of cirrhosis reversion. AB - Adenoviral vectors efficiently target normal liver cells; however, a clear-cut description of the safety boundaries for using adenovectors in hepatic cirrhosis has not been settled. With this in mind, we used a first-generation, replication deficient adenoviral vector carrying the E. coli lacZ gene (Ad5betaGal) to monitor therapeutic range, biodistribution, toxicity and transduction efficiency in Wistar rats made cirrhotic by two different experimental approaches resembling alcoholic cirrhosis and biliary cirrhosis in humans. Further, we show proof of concept on fibrosis reversion by a 'therapeutic' Ad-vector (AdMMP8) carrying a gene coding for a collagen-degrading enzyme. Dose-response experiments with Ad5betaGal ranging from 1 x 10(8)-3 x 10(12) viral particles (vp) per rat (250 g), demonstrated that adenovirus-mediated gene transfer via iliac vein at 3 x 10(11 )vp/rat, resulted in an approximately 40% transduction in livers of rats made cirrhotic by chronic intoxication with carbon tetrachloride, compared with approximately 80% in control non-cirrhotic livers. In rats made cirrhotic by bile duct obstruction only, 10% efficiency of transduction was observed. Biodistribution analyses showed that vector expression was detected primarily in liver and at a low level in spleen and kidney. Although there was an important increase in liver enzymes between the first 48 h after adenovirus injection in cirrhotic animals compared to non-transduced cirrhotic rats, this hepatic damage was resolved after 72-96 h. Then, the cDNA for neutrophil collagenase, also known as Matrix Metalloproteinase 8 (MMP8), was cloned in an Ad-vector and delivered to cirrhotic rat livers being able to reverse fibrosis in 44%. This study demonstrates the potential use of adenoviral vectors in safe transient gene therapy strategies for human liver cirrhosis. PMID- 11857070 TI - Targeted beta-globin gene conversion in human hematopoietic CD34(+ )and Lin( )CD38(-)cells. AB - Chimeric oligonucleotides have been used successfully to correct point and frameshift mutations in several cell types, as well as in animal and plant models. However, their application to primitive human blood cells has been limited. In this investigation, chimeric oligonucleotides designed to direct a site-specific nucleotide exchange in the human beta-globin gene (an A to T substitution within codon 6) were introduced into normal human CD34(+) and Lin( )CD38(-) cells via microinjection. This A to T nucleotide exchange introduces the single site mutation responsible for sickle cell anemia. In 23% of experimental samples, gene conversion was detected in the progeny of microinjected CD34(+) and Lin(-)CD38(-) cells that were cultured for at least 4 weeks. In addition, gene conversion was detected in the erythroid progeny of Lin(-)CD38(-) cells at the mRNA level. Conversion rates as high as 10-15% in 11% (five of 44) of experimental samples were confirmed by allele-specific PCR and sequence analysis of genomic DNA from the progeny of microinjected Lin(-)CD38(-) cells. Given that as few as 10% normal hematopoietic cells are sufficient to keep patients free of sickle cell disease, the level of conversion we have achieved in some samples may well be of therapeutic benefit in patients with sickle cell disease. PMID- 11857072 TI - Viral vector-mediated gene expression in olfactory ensheathing glia implants in the lesioned rat spinal cord. AB - Implantation of olfactory ensheathing glia (OEG) is a promising strategy to augment long-distance regeneration in the injured spinal cord. In this study, implantation of OEG following unilateral hemisection of the dorsal cervical spinal cord was combined with ex vivo gene transfer techniques. We report, to our knowledge for the first time, that purified cultures of primary OEG are capable of expressing a foreign gene following adenoviral (AdV) and lentiviral (LV) vector-mediated gene transfer. OEG implants subjected to AdV vector-mediated gene transfer expressed high levels of transgenic protein in both intact and lesioned spinal cord at 7 days after implantation. However, the levels of transgene expression gradually declined between 7 and 30 days after implantation in lesioned spinal cord. Infection with LV vectors resulted in stable transduction of primary OEG cultures and transgene expression persisted for at least 4 months after implantation. Genetic engineering of OEG opens the possibility of expressing additional neurotrophic genes and create optimal 'bridging' substrates to support spinal axon regeneration. Furthermore, stable transduction of OEG allows us to reliably study the behaviour of implanted cells and to obtain better understanding of their regeneration supporting properties. PMID- 11857073 TI - Efficacy of recombinant adenovirus as vector for allergen gene therapy in a mouse model of type I allergy. AB - DNA-based immunization represents an attractive alternative approach to the current treatment of allergic diseases by specific immunotherapy with allergen extracts. In this study, we used a replication-deficient adenovirus vector (AdCMV), to examine the in vivo efficacy of preventive and therapeutic genetic immunization in a mouse model of type I allergy. Primary immunization with a recombinant adenovirus expressing the model antigen beta-galactosidase (AdCMV (beta)gal) induced a Th1 immune response (predominance of IgG2a antibodies, high frequency of IFN-gamma producing T cells) and large numbers of cytotoxic T lymphocytes. Prophylactic vaccination with AdCMV-(beta)gal abolished the production of specific IgE following subsequent immunization with (beta)gal protein, and skewed the Th2-biased immune response to a Th1-orientated response. In contrast, therapeutic administration of AdCMV-(beta)gal after priming with (beta)gal-protein neither significantly inhibited ongoing IgE production nor modulated a manifest Th2 immune response. Thus, allergen gene transfer via recombinant adenovirus represents an effective method to establish protection against the development of allergic disorders, but does not qualify as a therapeutic tool to interfere with ongoing high IgE production. PMID- 11857075 TI - Concomitant inactivation of p53 and Chk2 in breast cancer. AB - The structure and expression of the human Rad53 homologue Chk2 was analysed in breast cancer. The previously described silent polymorphism at nucleotide 252 in codon 84 (GAA>GAG) was observed in 5/141 cases. Somatic Chk2 coding mutations were detected in 7/141 cases, these occurring in 4/18 BRCA1-associated breast cancers, 1/78 sporadic breast cancers and 2/25 typical medullary carcinomas. Each of the BRCA1-associated cancers with Chk2 mutations also contained mutations in p53, whereas the single sporadic cancer with Chk2 mutation was wild-type for p53. Expression of Chk2 was ubiquitously detected in normal ductal epithelium of the breast, but there was loss of expression in a significant proportion of breast carcinomas, and this occurred in cancers both with and without p53 mutation. A CpG island was identified 5' of the Chk2 transcriptional start site, but there was no evidence of cytosine methylation in any of the cancers with down-regulated Chk2 expression. Analysis of the germ-line of 45 individuals with hereditary or early onset breast cancer revealed wild-type Chk2 sequence in all cases. Thus, despite the rarity of somatic mutations in Chk2 in sporadic breast carcinomas, our results nevertheless reveal that concomitant loss of function in Chk2 (via down-regulation of expression) and p53 (via mutation) occurs in a proportion of sporadic cases. However, consistent with other studies, we show that germ-line mutations in Chk2 are unlikely to account for a significant proportion of non BRCA1-, non BRCA2-associated hereditary breast cancers. PMID- 11857074 TI - The LIM-domain protein Lmo2 is a key regulator of tumour angiogenesis: a new anti angiogenesis drug target. AB - The growth of solid tumours requires a blood supply provided by re-modeling of existing blood vessel endothelium (angiogenesis). Little is known about transcription regulators which are specific for the control of tumour angiogenesis. The proto-oncogene LMO2 encodes a LIM domain transcription regulator which controls angiogenesis during mouse embryogenesis where it regulates remodelling of the capillary network into mature vessels. We now show that Lmo2 expression is augmented in tumour endothelium such as mouse thymomas and human lung tumours. The functional significance of this Lmo2 expression was assessed in teratocarcinomas induced in nude mice by subcutaneous implantation of Lmo2-lacZ targeted ES cells. CD31-positive, sprouting endothelium of ES-cell origin occurred in teratocarcinomas from heterozygous Lmo2-lacZ ES cells but none occurred from null Lmo2-lacZ ES cells. Therefore, in this model Lmo2 is an obligatory regulator of neo-vascularization of tumours. These data suggest that LMO2 function may be a drug target in cancer and other conditions characterized by neo-vascularization. PMID- 11857076 TI - Astrocyte-specific expression of CDK4 is not sufficient for tumor formation, but cooperates with p53 heterozygosity to provide a growth advantage for astrocytes in vivo. AB - The development of malignant gliomas (astrocytomas) involves the accumulation of multiple genetic changes, including mutations in the p53 and retinoblastoma (Rb) cell cycle regulatory pathways. One Rb pathway alteration seen in high-grade astrocytomas is amplification of cyclin dependent kinase-4 (CDK4). To define the function of CDK4 amplification/overexpression in astrocytoma pathogenesis, we generated three transgenic mouse lines that overexpress human CDK4 (hCDK4) in astrocytes using the human glial fibrillary acidic protein (GFAP) promoter. GFAP hCDK4 mice do not develop brain tumors, but exhibit a small increase in astrocyte number. Cultured astrocytes from these mice do not demonstrate a cell-autonomous growth advantage in vitro and lack properties of transformed cells. To determine whether cdk4 overexpression provides a cooperative growth advantage in vitro, CDK4-overexpressing C6 glioma cell lines were generated and found to exhibit increased cell growth. In addition, GFAP-hCDK4; p53+/- as well as p53+/-; Rb+/- mice exhibited increased numbers of astrocytes compared to GFAP-hCDK4, p53+/-, or Rb+/- mice in vivo. No cooperative effect was observed with GFAP-hCDK4; Rb+/- mice. These results support the hypothesis that cdk4 overexpression alone is not sufficient for astrocytoma formation, but can provide a cooperative growth advantage in concert with genetic alterations in the p53 pathway. PMID- 11857077 TI - Synergy between truncated c-Met (cyto-Met) and c-Myc in liver oncogenesis: importance of TGF-beta signalling in the control of liver homeostasis and transformation. AB - The c-Met tyrosine kinase receptor and its ligand, Hepatocyte Growth Factor/ Scatter Factor, have been implicated in human cancer. We have previously described that the transgenic expression of a truncated form of human c-Met (cyto Met) in the liver confers resistance to several apoptotic stimuli. Here we show the impact of cyto-Met expression on liver proliferation and transformation. Despite a sixfold increase of hepatocyte proliferation, adult transgenic livers displayed normal size and architecture. We present evidence showing that activation of TGF-beta1 signalling controls the liver mass in cyto-Met mice. The oncogenic potential of cyto-Met was further assessed in the context of c-Myc induced hepatocarcinogenesis, using WHV/c-Myc transgenic mice. Co-expression of cyto-Met and c-Myc further enhanced hepatocyte proliferation and caused a dramatic acceleration of the Myc-induced tumorigenesis, leading to the emergence of hepatocarcinomas in 3-4-month-old animals. Importantly, the TGF-beta receptor type II expression was strongly downregulated in most tumours, indicating that impairment of TGF-beta1-mediated growth inhibition plays a major role in accelerated neoplastic development. The strong potential of cyto-Met for oncogenic cooperation without direct transforming activity designates cyto-Met mice as an ideal tool for studying the early steps of multistage hepatocarcinogenesis and for identification of prognostic markers of transformation. PMID- 11857078 TI - Identification of genes regulated by dexamethasone in multiple myeloma cells using oligonucleotide arrays. AB - Our previous studies have characterized Dexamethasone (Dex)-induced apoptotic signaling pathways in multiple myeloma (MM) cells; however, related transcriptional events are not fully defined. In the present study, gene expression profiles of Dex-treated MM cells were determined using oligonucleotide arrays. Dex triggers early transient induction of many genes involved in cell defense/repair-machinery. This is followed by induction of genes known to mediate cell death and repression of growth/survival-related genes. The molecular and genetic alterations associated with Dex resistance in MM cells are also unknown. We compared the gene expression profiles of Dex-sensitive and Dex-resistant MM cells and identified a number of genes which may confer Dex-resistance. Finally, gene profiling of freshly isolated MM patient cells validates our in vitro MM cell line data, confirming an in vivo relevance of these studies. Collectively, these findings provide insights into the basic mechanisms of Dex activity against MM, as well as mechanisms of Dex-resistance in MM cells. These studies may therefore allow improved therapeutic uses of Dex, based upon targeting genes that regulate MM cell growth and survival. PMID- 11857079 TI - Direct repression of the Mcl-1 promoter by E2F1. AB - E2F1 induces apoptosis via both p53-dependent and p53-independent mechanisms. The direct targets in the p53-independent pathway remain enigmatic; however, the induction of this pathway does not require the transactivation domain of E2F1. Using cells that are defective in p53 activation, we show that E2F1 potently represses the expression of Mcl-1--an anti-apoptotic Bcl-2 family member whose depletion results in apoptosis. We also show that this transcriptional repression is direct and dependent upon E2F1's DNA-binding domain, but does not require the transactivation domain of E2F1. Consistent with this DNA binding requirement of E2F1 to repress Mcl-1, we show that E2F1 binds to the Mcl-1 promoter both in vitro and in vivo, and have identified the DNA element (-143/-117) within this promoter that is required for E2F1 binding and repression. Additionally, cell lines constitutively expressing Mcl-1 are resistant to E2F1-mediated apoptosis- suggesting that Mcl-1 downregulation is a necessary event in the p53-independent apoptotic process. Thus, we identify a p53 family-independent mechanism of E2F1 induced apoptosis in which E2F1 directly represses Mcl-1 expression. PMID- 11857080 TI - Direct integrin alphavbeta6-ERK binding: implications for tumour growth. AB - Blockade of the mitogen-activated protein (MAP) kinase pathway suppresses growth of colon cancer in vivo. Here we demonstrate a direct link between the extracellular signal-regulated kinase ERK2 and the growth-promoting cell adhesion molecule, integrin alphavbeta6, in colon cancer cells. Down-regulation of beta6 integrin subunit expression inhibits tumour growth in vivo and MAP kinase activity in response to serum stimulation. In alphavbeta6-expressing cells ERK2 is bound only to the beta6 subunit. The increase in cytosolic MAP kinase activity upon epidermal growth factor stimulation is all accounted for by beta6-bound ERK. Deletion of the ERK2 binding site on the beta6 cytoplasmic domain inhibits tumour growth and leads to an association between ERK and the beta5 subunit. The physical interaction between integrin alphavbeta6 and ERK2 defines a novel paradigm of integrin-mediated signalling and provides a therapeutic target for cancer treatment. PMID- 11857081 TI - The putative tumor suppressor RASSF1A homodimerizes and heterodimerizes with the Ras-GTP binding protein Nore1. AB - Nore and RASSF1A are noncatalytic proteins that share 50% identity over their carboxyterminal 300 AA, a segment that encompasses a putative Ras-Rap association (RA) domain. RASSF1 is expressed as several splice variants, each of which contain an RA domain, however the 340 AA RASSF1A, but not the shorter RASSF1C variant, is a putative tumor suppressor. Nore binds to Ras and several Ras-like GTPases in a GTP dependent fashion however neither RASSF1 (A or C) or the C. elegans Nore/RASSF1 homolog, T24F1.3 exhibit any interaction with Ras or six other Ras-like GTPases in a yeast two-hybrid expression assay. A low recovery of RASSF1A (but not RASSF1C) in association with RasG12V is observed however on transient expression in COS cells. Nore and RASSF1A can each efficiently homodimerize and heterodimerize with each other through their nonhomologous aminoterminal segments. Recombinant RASSF1C exhibits a much weaker ability to homodimerize or heterodimerize; thus the binding of RASSF1C to Nore is very much less than the binding of RASSF1A to Nore. The association of RASSF1A with RasG12V in COS cells appears to reflect the heterodimerization of RASSF1A with Nore, inasmuch the recovery of RASSF1A with RasG12V is increased by concurrent expression of full length Nore, and abolished by expression of Nore deleted of its RA domain. The preferential ability of RASSF1A to heterodimerize with Nore and thereby associate with Ras-like GTPases may be relevant to its putative tumor suppressor function. PMID- 11857083 TI - A single mutated BRCA1 allele leads to impaired fidelity of double strand break end-joining. AB - Heterozygosity for mutations in the BRCA1 gene in humans confers high risk for developing breast cancer, but a biochemical basis for this phenotype has not yet been determined. Evidence has accumulated implicating BRCA1, in the maintenance of genomic integrity and the protection of cells against DNA double strand breaks (DSB). Here we present evidence that human cells heterozygous for BRCA1 mutations exhibit impaired DNA end-joining, which is the major DSB repair pathway in mammalian somatic cells. Using an in vivo host cell end-joining assay, we observed that the fidelity of DNA end-joining is strongly reduced in three BRCA1(+/-) cell lines in comparison to two control cell lines. Moreover, cell free BRCA1(+/-) extracts are unable to promote accurate DNA end-joining in an in vitro reaction. The steady-state level of the wild type BRCA1 protein was significantly lower than the 50% expected in BRCA1(+/-) cells and thus may underlie the observed end-joining defect. Together, these data strongly suggest that BRCA1 is necessary for faithful rejoining of broken DNA ends and that a single mutated BRCA1 allele is sufficient to impair this process. This defect will compromise genomic stability in BRCA1 germ-line mutation carriers, triggering the genetic changes necessary for the initiation of neoplastic transformation. PMID- 11857082 TI - Role of the AKT kinase in expansion of multiple myeloma clones: effects on cytokine-dependent proliferative and survival responses. AB - IL-6 is an established growth factor for multiple myeloma tumor cells, stimulating proliferative and survival responses. Recent work indicates that IL-6 can activate the AKT kinase in myeloma cells. Thus, to test a potential role for AKT in IL-6-induced cellular responses, we transfected myeloma cell lines with an active 'E40K' or dominant negative'PH AKT construct using an adenoviral vector. Transfection of the E40K into myeloma cells resulted in enhanced tumor cell growth and expression of the PH dominant negative AKT resulted in both inhibition of the IL-6-dependent proliferative response and a decrease in S phase distribution. While transfection of E40K protected myeloma cells from dexamethasone-induced apoptosis, the dominant negative PH had no effect on the ability of IL-6 to protect these cells from dexamethasone. These results clearly demonstrate that AKT activation is critical for the IL-6 proliferative response. In addition, although the level of AKT activation can regulate sensitivity to dexamethasone-induced apoptosis, additional cytokine-induced AKT-independent pathways can mediate IL-6 protection against dexamethasone. DOI: 10.1038/sj/onc/1205194 PMID- 11857084 TI - Adenovirus E1A requires the yeast SAGA histone acetyltransferase complex and associates with SAGA components Gcn5 and Tra1. AB - The budding yeast Saccharomyces cerevisiae was used as a model system to study the function of the adenovirus E1A oncoprotein. Previously we demonstrated that expression of the N-terminal 82 amino acids of E1A in yeast causes pronounced growth inhibition and specifically interferes with SWI/SNF-dependent transcriptional activation. Further genetic analysis identified the yeast transcription factor Adr1 as a high copy suppressor of E1A function. Transcriptional activation by Adr1 requires interaction with co-activator proteins Ada2 and Gcn5, components of histone acetyltransferase complexes including ADA and SAGA. Analysis of mutant alleles revealed that several components of the SAGA complex, including proteins from the Ada, Spt, and Taf classes were required for E1A-induced growth inhibition. Growth inhibition also depended on the Gcn5 histone acetyltransferase, and point mutations within the Gcn5 HAT domain rendered cells E1A-resistant. Also required was SAGA component Tra1, a homologue of the mammalian TRRAP protein which is required for c-myc and E1A induced cellular transformation. Additionally, Gcn5 protein could associate with E1A in vitro in a manner that depended on the N-terminal domain of E1A, and Tra1 protein was co-immunoprecipitated with E1A in vivo. These results indicate a strong requirement for intact SAGA complex for E1A to function in yeast, and suggest a role for SAGA-like complexes in mammalian cell transformation. PMID- 11857085 TI - Differential expression and signaling of CBL and CBL-B in BCR/ABL transformed cells. AB - CBL and the related CBL-B protein are two members of a family of RING finger type ubiquitin E3 ligases that are believed to function as negative regulators of signal transduction in hematopoietic and immune cells. In mice, expression of v Cbl causes lymphomas, and targeted disruption of either the CBL gene or the CBL-B gene can result in a lymphoproliferative disorder or hypersensitivity of lymphocytes. CBL is one of the most prominent targets of the BCR/ABL tyrosine kinase oncogene. We compared the role of CBL and CBL-B in signal transduction of BCR/ABL using pairs of cell lines before and after expression of BCR/ABL. In contrast to CBL, BCR/ABL was found to rapidly downregulate the expression of CBL B protein. The decrease in CBL-B protein induced by BCR/ABL was associated with downregulation of CBL-B mRNA. Downregulation and tyrosine phosphorylation of CBL B required BCR/ABL kinase activity. However, despite their known similarities in structure and function, we found CBL and CBL-B proteins to be involved in distinct signaling complexes. CBL was predominantly in a complex with phosphatidylinositol 3'-kinase and CRKL, while CBL-B was not associated with any significant phosphatidylinositol 3'-kinase activity. A major CBL-B associated protein was identified as mono-ubiquitinated Vav, a nucleotide exchange factor for Rac1. These results demonstrate that BCR/ABL signals differentially through CBL and CBL-B, with downregulation of the CBL-B protein potentially contributing to BCR/ABL-mediated transformation. PMID- 11857086 TI - SV40 infection induces telomerase activity in human mesothelial cells. AB - Mesotheliomas are malignant tumors of the pleural and peritoneal membranes which are often associated with asbestos exposure and with Simian virus 40 (SV40) infection. Telomerase activity is repressed in somatic cells and tissues but is activated in immortal and malignant cells. We evaluated telomerase activity in seven primary malignant mesothelioma biopsies and matched lung specimens and 20 mesothelioma cell lines and eight corresponding primary tumor cultures. All the tumor biopsies, and nearly all primary cell mesothelioma cultures and cell lines were telomerase positive. The findings in cell lines paralleled those observed in primary cultures in cases where paired samples were available. Next, we found that SV40, a DNA tumor virus present in approximately 50% of mesothelioma biopsies in the USA, induced telomerase activity in primary human mesothelial cells, but not in primary fibroblasts. Telomerase activity became detectable as early as 72 h following wild-type (strain 776) SV40 infection, and a clear DNA ladder was detectable 1 week after infection. The amount of telomerase activity increased during passage in cell culture and appeared to parallel increases in the cellular amounts of the SV40 large T-antigen. Thus, SV40 infection leads to telomerase activity before the infected mesothelial cells become transformed and immortalized. SV40 infection of human fibroblasts did not cause detectable telomerase activity. We also determined that the SV40 small t-antigen (tag) plays an important role in inducing telomerase activity because this activity was undetectable or minimal in mesothelial cells infected and/or transformed by SV40 tag mutants. Asbestos alone did not induce telomerase activity, and asbestos did not influence telomerase activity in mesothelial cells infected with SV40. Induction of telomerase activity by SV40 may be related to the very high rate of mesothelial cell immortalization that is characteristically associated with SV40 infection of mesothelial cells. PMID- 11857087 TI - Chromosomal autonomy of hMLH1 methylation in colon cancer. AB - Silencing of hMLH1 expression by aberrant hMLH1 promoter methylation accounts for the majority of sporadic colon cancers with microsatellite instability. We have previously shown hMLH1 silencing is biallelic and actively maintained. To study the mechanism of aberrant hMLH1 methylation, we assayed whether an hMLH1 methylated cell could transfer methylation and silencing to an exogenous hMLH1 promoter in somatic cell hybrids between hMLH1 methylated-silenced and hMLH1 unmethylated-expressing colon cancer cells. Conversely, we assayed whether these hybrids could reactivate expression of initially methylated and silenced hMLH1 alleles. Compellingly, within the hybrids each hMLH1 allele remained unchanged, retaining the expression status of its parental cell of origin. This chromosomal autonomy may not be simply determined by DNA methylation, as it is reasserted after experimentally forced demethylation of all hMLH1 alleles in the hybrids. Confirming findings included hMLH1 methylated cells being unable to methylate single transferred exogenous hMLH1 expressing chromosomes or transfected hMLH1 reporter constructs. hMLH1 silencing does not conform to either a dominant or recessive model, and is not determined by trans-acting factors differing between hMLH1 expressing or silenced genomes. We posit that hMLH1 methylation is dependent on and maintained by cis chromosomal marks, whose nature remains to be elucidated. PMID- 11857089 TI - Identification of a region of homozygous deletion on 8p22-23.1 in medulloblastoma. AB - To identify critical tumor suppressor loci that are associated with the development of medulloblastoma, we performed a comprehensive genome-wide allelotype analysis in a series of 12 medulloblastomas. Non-random allelic imbalances were identified on chromosomes 7q (58.3%), 8p (66.7%), 16q (58.3%), 17p (58.3%) and 17q (66.7%). Comparative genomic hybridization analysis confirmed that allelic imbalances on 8p, 16q and 17p were due to loss of genetic materials. Finer deletion mapping in an expanded series of 23 medulloblastomas localized the common deletion region on 8p to an interval of 18.14 cM on 8p22-23.2. We then searched within the region of loss on 8p for loci that might contain homozygous deletion using comparative duplex PCR. An overlapping homozygous deletion region was identified in a 1.8 cM interval on 8p22-23.1, between markers D8S520 and D8S1130, in two medulloblastomas. This region of homozygous deletion also encompasses the 1.4 cM minimal deletion region detected on 8p in ductal carcinoma in situ of breast. In conclusion, we reported for the first time a detailed deletion mapping on 8p in medulloblastoma and have identified a region of homozygous deletion on 8p22-23.1 that is likely to contain a critical tumor suppressor gene involved in the development of medulloblastoma. PMID- 11857088 TI - PTEN regulates tumor cell adhesion of colon carcinoma cells under dynamic conditions of fluid flow. AB - The regulation of integrin-mediated cell adhesion and its stabilization involves different phosphorylation and dephosphorylation events. Focal adhesion kinase (FAK) has been recently found to be a substrate of the dual-specific phosphatase PTEN in glioma cells, where it appears to be involved in regulation of cell spreading and migration as part of focal adhesions. We have investigated the role of PTEN in cell adhesion of HT-29 human colon carcinoma cells under static and hydrodynamic conditions of fluid flow. PTEN coprecipitated with FAK and paxillin dependent on the formation of adhesions to collagens. This corresponded with an adhesion-dependent increase in Tyr-phosphatase activity of PTEN. Using preparations of native FAK and PTEN from HT-29 cells in a specific Tyr phosphatase assay FAK was identified as substrate for this dephosphorylation. If expression of PTEN was reduced using antisense oligonucleotides cell adhesion under dynamic conditions of laminar flow, but not under static conditions was significantly increased. In addition, cell spreading was increased in cells with reduced PTEN expression. We conclude that PTEN appears to be involved in the regulation of integrin-mediated adhesion through dephosphorylation of FAK. This phosphatase might play a role as a negative regulator for the formation of stable HT-29 cell adhesion to extracellular matrix. PMID- 11857090 TI - Impact of advanced maternal age on pregnancy outcome. AB - The aim of this study was to compare the pregnancy outcome and delivery complications in women 40 years or older (cases) to that of women 20 to 30 years old (controls). Over a 5-year period, 319 cases had a singleton delivery in our institution. These women were compared with 326 controls. Parity was significantly higher in cases compared with controls (3.2 vs. 1.8). Advanced maternal age, compared with younger age, was associated with significantly higher rates of preterm delivery (16.0 vs. 8.0%), cesarean delivery (CS) (31.3 vs. 13.5%), and the occurrence of one or more antepartum complications (29.5 vs. 16.6%). When the two groups were subdivided according to parity, rates of preterm delivery, CS, preeclampsia, gestational diabetes, chronic hypertension, and labor induction were each significantly higher among older multiparas compared with control multiparas. However, only preterm delivery, CS rates, and uterine fibroids were found to be significantly higher in older nulliparous compared with young nulliparous women. We conclude that multiparous women at least 40 years old have a higher antepartum complication rate including intrauterine fetal death compared with younger women. PMID- 11857091 TI - Contemporary trends in the management of delivery at 23 weeks' gestation. AB - The objective of this study is to investigate the current understanding of neonatal survival and the willingness to provide aggressive obstetric intervention at the limit of fetal viability among practicing perinatologists in the United States. A pretested survey was mailed to members of the Society for Maternal Fetal Medicine. The survey identified the practitioner's opinion of the lowest limit of viability and gestational ages at which antenatal steroids and cesarean section would first be provided. We also attempted to identify practitioner knowledge of survival at 23 and 24 weeks' gestation and years of perinatal practice. Of the 1244 surveys mailed to members of the Society for Maternal-Fetal Medicine practicing in the United States, 462 practitioners replied for a 37% response rate. Fifty percent considered the lower limit of viability to be 24 weeks' and 44.3% believed the lower limit of viability to be 23 weeks' gestation. Among respondents, the majority estimated a 0 to 10% survival rate at 23 weeks' and 25 to 50% survival rate at 24 weeks. Only 13% of respondents correctly estimates survival at 23 weeks' gestation as >25%, and only 17% correctly estimated survival at 24 weeks' at >50%. Among practitioners, 43.6% would first give antenatal steroids after 24 weeks' gestation but 55.6%would administer steroids before the beginning of week 24. Twenty-eight percent would routinely monitor a fetus less than 24 weeks' gestation. Consistent with this observation, 21% would perform a cesarean section before 24 weeks' gestation. We find that respondents underestimate the consensus in the most recent literature on survival of infants born at 23 to 24 weeks. This underestimation may lead to a limitation in interventions offered. PMID- 11857092 TI - Neonatal clavicular fracture: clinical analysis of incidence, predisposing factors, diagnosis, and outcome. AB - The objective of this study is to identify maternal, perinatal, and fetal risk factors for clavicular fracture in a single institution. We performed a prospective study of all deliveries during a 14-month period to identify confirmed cases of neonatal clavicular fracture. The control group consisted of the deliveries immediately preceding and following the index cases. Fifty-three cases of clavicular fracture were identified among the 4789 deliveries from October 1995 through November 1996 for an incidence of 1.11%. Three neonates in the clavicular fracture group were delivered through cesarean section. Neonates with fracture were significantly heavier at birth than those without (3564 vs. 3283 g, p <0.001), and had a lower mean head-to-abdominal circumference ratio (0.93 vs. 1.08, p <0.001), history of giving birth to a macrosomia (21 vs. 4%, p <0.05). The anterior shoulder was the predominant site of fracture (30/53). Fracture was detected mostly during the first 3 days of neonatal life (46/53). The outcome was benign, with complete recovery in all cases and no associated neurological sequelae. Neonatal clavicular fracture tended to be associated with neonatal somatometric characteristics and difficult deliveries. Considering the benign nature of this birth trauma, more invasive intrapartum management to lower its incidence is not advised. PMID- 11857093 TI - Calcium and phosphorus balance of extremely preterm infants with estradiol and progesterone replacement. AB - Infants born extremely prematurely are deprived of the placental supply of estradiol (E2) and progesterone (Prog) at an earlier developmental stage compared to an infant born at term. We hypothesized that the retention of Ca (calcium) and P (phosphorus) would be improved by an E2 and Prog replacement. Twenty female infants with a mean gestational age of 26.6 weeks (+/-1.5 SD) and a mean birth weight of 744 g (+/-156) were enrolled in a randomized controlled pilot study. One group received an E2 and Prog replacement to maintain intrauterine plasma concentrations of E2 and Prog and the other group served as control. When intake of formula was at least 100 mL/kg/d, a 3-day Ca and P balance study was performed. Ca and P intake was increased individually until both elements were excreted in the urine. The mean Ca and P retention was 4.21 (+/-1.75) mMol/kg/d (58% of intake) and 2.66 (+/-1.01) mMol/kg/d (80%) in the replaced group and 3.39 (+/-1.69) mMol/kg/d (56%) and 2.03 (+/-0.79) mMol/kg/d (71%) in the control group, respectively. In this pilot study the retention of Ca and P was not improved by an E2 and Prog replacement. PMID- 11857094 TI - Diagnosis and management of primary aldosteronism in pregnancy: case report and review of the literature. AB - Primary aldosteronism rarely complicates pregnancy. We present a woman with primary aldosteronism in pregnancy associated with severe preeclampsia. A 33-year old Japanese woman with hypertension was referred to our hospital at 25 weeks of gestation. Her blood pressure was 180/100 mmHg, and laboratory tests identified a low serum potassium level and moderate proteinuria on urinalysis. The fetus was diagnosed with growth restriction. Plasma renin activity (PRA) value was 2.2 ng/mL/h and plasma aldosterone concentration (PAC) was elevated (260 pg/mL). The patient was treated medically. At 27 weeks of gestation, we noted persistent late fetal heart rate decelerations associated with uterine contractions. Therefore, elective caesarean section was performed and she was delivered of a 698-g female. After delivery, PRA declined and PAC remained elevated. Abdominal computerized tomography scan and I131-iodochoresterol scan revealed a tumor in the left adrenal gland. Laparoscopic adrenalectomy was performed and confirmed the clinical diagnosis. PMID- 11857095 TI - Obstetric implications of the factor V leiden mutation: a review. AB - Factor V Leiden (FVL) is a newly discovered genetic mutation that impairs one of the body's naturally occurring anticoagulation systems. The result is resistance to activated protein C and a predisposition to thrombosis. FVL is the most common cause of primary and recurrent venous thromboembolism in the pregnant and nonpregnant state. The FVL gene is common in the general population and transmitted in an autosomal dominant fashion. When FVL is combined with the prothrombotic state of pregnancy, the result is an increased propensity to manifest a number of pregnancy complications. These include recurrent pregnancy loss and stillbirth, severe and early-onset preeclampsia, placental abruption and possibly, intrauterine growth restriction. It remains unknown whether thromboprophylaxis is effective in ameliorating these pregnancy complications. The current literature and management recommendations are highlighted in this article. PMID- 11857096 TI - Effects of maternal and paternal age on Caucasian and Native American preterm births and birth weights. AB - The purpose of this study was to compare the effects of maternal and paternal age on Native American and Caucasian infants. Data were abstracted from birth records compiled in North Dakota from 1978 to 1992. Native Americans had a 45% higher risk for preterm births, a 140% higher risk for low birth weight (LBW) (<2500 g), and a 131% higher risk for very LBW (<1500 g) compared with Caucasians. Maternal and paternal age both had a "U-shaped" effect on preterm birth. However, while the risk for preterm birth was increased for both teenage mothers and fathers, the risk was also significantly increased only for mothers between the ages of 36 40 and 41-45. There was also a "U-shaped" relationship between LBW and parental age, and again the risk was statistically significant for different age groups. For mothers, the risk was decreased significantly for those 26 to 30 years of age, whereas for fathers, it was increased significantly for those <20 years of age. There was also a significant interaction between paternal age and race, with teenage Native American fathers having a slight (4%), but statistically significant, increased risk for having LBW children compared with teenage Caucasian fathers. The relationship between parental age and very LBW was also "U shaped," but while the odds ratios for each maternal age group were significant, none of the odds ratios for any of the paternal age groups was statistically significant. The influence of teenage fathers on preterm and LBWs has not been previously reported, and the finding that paternal age has a greater influence among Native Americans is especially intriguing and deserving of further investigation. PMID- 11857097 TI - Air emboli in the intracranial venous sinuses of neonates. AB - Air bubbles in the intracranial venous sinuses are known as a consequence to different causes including trauma, infection, and administration of intravenous contrast. Most of the previous reports demonstrated such cases in adults, with subsequent complications. We are presenting two premature babies who developed asymptomatic air bubbles in the right cavernous and left transverse sinuses, introduced accidentally upon cannulation of scalp veins. In both babies the air embolism disappeared in a few days without complications. Our cases suggest that these accidents could happen more frequently in neonates following scalp vein cannulation, which is a common procedure in sick babies, but they were overlooked as the outcome was uneventful. However, the precise nature and clinical significance of this lesion is not well understood in neonates. PMID- 11857098 TI - [Surgery and long-term follow-up of hepatic echinococcosis outside endemic regions]. AB - The aim of the study was to characterize the therapeutic pathways in patients with echinococcal hydatids in an institution outside the endemic areas but with a high frequency of oncological hepatic surgery. PATIENTS AND METHODS: 44 patients with echinococcosis from 1987-1997 were reviewed. To evaluate the long-term results, clinical examination, chest x-ray, serology and liver ultrasound were performed. According to the study protocol 39 patients were evaluable. 89.7 % of the patients suffered from a primary disease, whereas 7.7 had a local recurrence and another 2.5 of patients showed a second site of manifestation after former operation. E. cysticus occurred in 82.5 %, E. alveolaris in 17.5 %. RESULTS: In 66.7 % of all cases a resecting procedure (pericystectomy, atypic resection, regular and extended hemihepatectomy) was performed. One patient underwent an orthotopic liver transplantation and in 10 % the surgical procedure included only an open drainage of the cysts. A simple cystectomy was performed in 23 %. One patient died from multiple organ failure as a consequence of local bleeding complications. The overall complication rate was 38.5 %, including the postoperative death and 7.7 % reoperations. The complication rate following pericystectomy was much higher than after simple cystectomy. At the time of follow-up (median 66 months) no patient showed a recurrent disease. CONCLUSION: Compared to reports from endemic regions the rate of resective procedures was much higher. The therapeutic strategy lead to excellent long-term results. Simple cystectomy should be preferred as pericystectomy showed a higher morbidity. PMID- 11857099 TI - [5-Fluorouracil-induced colitis--a review based upon consideration of 6 cases]. AB - BACKGROUND: At increasing use of high-dose 5-fluorouracil-based chemotherapy for metastatic colorectal and gastric cancer complicated drug-induced colitis is observed more frequently. From May 1998 to November 2000 we observed 6 cases of 5 fluorouracil-induced colitis, in which we looked for involvement of small intestine. We report summing up on the 6 cases including both endoscopic and histological findings in both sites of the gut. CASE REPORTS: In 2 men and 4 women (age 49-78 years) with advanced colon (n = 2), gastric (n = 3 ) and gallbladder (n = 1) cancer a palliative weekly high-dose infusional 5 fluorouracil (2,6 g/m(2)/24 h) and folinic acid (500 mg/m(2)/2 h) chemotherapy was performed. Few days after 1-5 chemotherapy courses the patients were admitted to our hospital with abdominal pain and partly severe watery diarrhea (up to 20 times evacuations/per day). The stool cultures were negative and there were no proof both of clostridium difficile and his toxin A and B. In 4 patients colonoscopy showed different grades of colitis up to diffuse erythema and microlesions, 2 patients had no visible lesions. In 4 patients endoscopy of the upper GI-tract showed a severe inflammation (n = 1) and a fibrinopurulent exsudate, severe edema and isolated ulcerations (n = 3) of jejunum after gastrectomy or duodenum with intact stomach. In the histological assessment different grades of 5-FU-induced colitis without (n = 2) or with (n = 4) involvement of the upper small intestine destruction of the superficial mucosa and crypts (epitheliumapoptosis) were found. 5 patients were treated by antibiotics (vancomycin n = 2, metronidazole n = 3), glucocorticoids (n = 5) and Saccaromyces cerevisiae (n = 3). After 8-10 days the patients were complete free of symptoms. One patient died due to the enterocolitis. CONCLUSIONS: The present cases demonstrate that high-dose 5-fluorouracil-based chemotherapy not only induces a colitis but also may involve the upper small intestine tract. Consequently, it represents an increasing and serious adverse event of high-dose chemotherapy. The etiology of the enterocolitis (drug- or bacterial-induced) needs further investigations in order to find a causal therapy and/or prophylaxis. PMID- 11857100 TI - Transanal endoscopic microsurgery (TEM) for minimally invasive resection of rectal adenomas and "Low-risk" carcinomas (uT1, G1 - 2). AB - Transanal endoscopic microsurgery (TEM) presents a minimally invasive procedure for local removal of large rectal adenomas (>/= 2 cm) and early, so called "low risk" carcinomas (uT1, G1 - 2) in curative as well as of advanced tumors in palliative intent. Over a 6-year period 92 TEM excisions of rectal tumors were carried out including 91 patients with 56 adenomas, 35 carcinomas (9 pTis, 17 pT1, 5 pT2, 3 pT3, 1 Ca after snare diathermy) and one neurinoma. Two patients of the carcinoma group had to be reoperated by means of anterior resection due to false preoperative rectal ultrasound examination (2 x uT1--> pT2). 4 patients required palliative therapy on account of age or high morbidity. After a mean follow-up time of 23 months (adenomas 23 months, pT1 carcinoma 26 months and advanced tumors 38 months) we encountered a total of 7 complications, of which in 5 cases surgical reintervention was necessary (5,4 %). One 86-year-old patient with a pT2-carcinoma, who was unsuitable for low anterior resection due to a high morbidity risk, died from myocardial infarction after emergency reintervention caused by postoperative bleeding. To date, overall 9 recurrences occurred (9,8 %). In the specific target group of TEM (adenomas and pT1, G1-2 carcinomas) consisting of n = 83 cases, the overall recurrence rate was 7,2 %, of which 5,3 % were due to adenomas and 11,5 % due to carcinomas. After palliative excision 2 recurrences occurred. These results of transanal endoscopic microsurgery (TEM) indicate that this technique has a useful place in curative, as well as in palliative management of rectal tumors. PMID- 11857101 TI - [Fever and weight loss as leading symptoms of infection with giardia lamblia]. AB - Infections, malignancies and autoimmune diseases are the most important causes of fever of unknown origin.A case report of a 80-year old patient is described, who was admitted to our hospital because of fever lasting more than 2 weeks, weight loss and lack of appetite. Physical examination did not provide any relevant information, laboratory tests revealed an elevation of inflammation markers and anemia. Serological tests for infectious and autoimmune diseases and cultures of stool, blood and urine were all negative. Imaging did not show any pathological findings, colonoscopy and gastroscopy were macroscopically normal. Surprisingly, histology showed massive giardiasis of the duodenum. After initiation of therapy with metronidazol, fever and inflammation markers declined and the patient could be discharged from hospital without complaints on the 15(th) day after admission. Giardia lamblia is one of the most common intestinal pathogens worldwide. Infection can cause acute diarrhea, but may also be responsible for chronic abdominal complaints or may stay asymptomatic. To our knowledge, giardiasis has not been described as differential diagnosis of fever of unknown origin so far. In synopsis of clinical presentation and outcome after antibiotic therapy, we postulate that Giardia lamblia was the relevant cause of fever and weight loss in this case. In patients presenting with these symptoms, Giardia lamblia should be considered as differential diagnosis. PMID- 11857102 TI - [Obstructive jaundice and acute pancreatitis due to an obstruction of the afferent loop after billroth-II-resection]. AB - An obstruction of the afferent loop after Billroth-II-resection is an extremely rare late complication of this procedure. We report on a 76-year-old female patient with a history of Billroth-II-resection 11 years ago who was admitted due to acute pancreatitis and obstructive jaundice. Abdominal sonography lead to the suspicion of a dilated afferent loop, which could be proven by means of magnetic resonance imaging. A tumorous lesion as cause of the obstructive jaundice was not detectable. Intraoperatively a volvulus of the small intestine and strangling adhesions near the Braun's anastomosis were seen, causing the obstruction of the afferent loop. Following reposition of the small intestine and adhesiolysis the patient gained a quick relief of symptoms and the jaundice disappeared completely. PMID- 11857103 TI - Fatal Hemorrhage following perforation of the aorta by a barb of the Gianturco Rosch esophageal stent. AB - Self-expanding metal stents are an established option in the palliative treatment of malignant stenoses of the esophagus. Herein, we report on a 60-year-old man with a recurrent stenosis that developed 2 months after radiochemotherapy for a squamous cell carcinoma in the middle part of the esophagus. To relieve progressive dysphagia, a Gianturco-Rosch stent (Cook-Z stent, 10 cm, PE-covered, manufactured by William Cook Europe) was implanted. Six weeks later, precipitous massive hemoptysis leading to the collapse and death of the patient occurred. Autopsy showed that a barb in the middle of the stent had perforated the aortic arch, resulting in massive bleeding into the gastrointestinal tract, and aspiration. Although hemorrhage and esophageal perforation are known late complications of all types of metal stents, our case is the first description of a perforation involving a fixation barb. These barbs are a particular feature of the European version of the Cook-Z stent, and are intended to prevent stent migration. In future, any hemorrhage observed after stent implantation should prompt a search for perforation by a barb (autopsy!). If necessary, the European version of the Gianturco Z stent should be modified. PMID- 11857104 TI - [Chronic intestinal pseudo-obstruction: pathogenesis, diagnosis and therapy]. AB - Chronic intestinal pseudo-obstruction (CIPO) is a rare disease in which a severe intestinal motility disorder impairs transit of chyme so that patients suffer from symptoms of a mechanical ileus without mechanical obstruction. CIPO may be a primary or secondary disorder due to muscular, neurologic, metabolic or endocrine disorders, but may also occur postinfectiously, postoperatively, following abdominal radiation or be caused by drugs or noxae. In severe cases, the typical history of (repeated) symptoms of mechanical obstruction leading to unsuccessful laparotomies will give key clues for diagnosis. If CIPO is suspected, mechanical obstruction must be searched for carefully by radiologic and endoscopic examinations. Histologic diagnosis usually demands full thickness biopsies of the intestinal wall. Small intestinal manometry allows diagnosis of CIPO even during oligosymptomatic intervals as well as differentiation between neuropathic and myopathic forms of the disease. The main therapeutic goals consist in: 1. Maintenance of an adequate nutritional state by oral and/or enteral nutrition; in severe cases home-parenteral nutrition may be required and particularly in children intestinal transplantation may be the ultima ratio. 2. Reconstitution of intestinal propulsion by prokinetic drugs. 3. Therapy of complications such as bacterial overgrowth and severe pain by antibiotics and specific surgical procedures. Unnecessary laparotomies should be strictly avoided because they may lead to adhesions and markedly complicate the clinical course. PMID- 11857105 TI - [Pancreatic Enzymes and intestinal Ischemia]. PMID- 11857106 TI - [Early gastrectomy in young, asymptomatic carriers of E-cadherin germline mutations]. PMID- 11857107 TI - [Long-term prophylaxis with omeprazol is superior to eradication therapy in H. pylori-positive NSAIDs taking patients]. PMID- 11857224 TI - Erratum. PMID- 11857108 TI - Premature chromosome condensation in humans associated with microcephaly and mental retardation: a novel autosomal recessive condition. AB - We report a novel autosomal recessive disorder characterized by premature chromosome condensation in the early G2 phase. It was observed in two siblings, from consanguineous parents, affected with microcephaly, growth retardation, and severe mental retardation. Chromosome analysis showed a high frequency of prophase-like cells (>10%) in lymphocytes, fibroblasts, and lymphoblast cell lines with an otherwise normal karyotype. (3)H-thymidine-pulse labeling and autoradiography showed that, 2 h after the pulse, 28%-35% of the prophases were labeled, compared with 9%-11% in healthy control subjects, indicating that the phenomenon is due to premature chromosome condensation. Flow cytometry studies demonstrate that the entire cell cycle is not prolonged, compared with that in healthy control subjects, and compartment sizes did not differ from those in healthy control subjects. No increased reaction of the cells to X-irradiation or treatments with the clastogens bleomycin and mitomycin C was observed, in contrast to results in the cell-cycle mutants ataxia telangiectasia and Fanconi anemia. The rates of sister chromatid exchanges and the mitotic nondisjunction rates were inconspicuous. Premature entry of cells into mitosis suggests that a gene involved in cell-cycle regulation is mutated in these siblings. PMID- 11857109 TI - X-linked cone-rod dystrophy (locus COD1): identification of mutations in RPGR exon ORF15. AB - X-linked cone-rod dystrophy (COD1) is a retinal disease that primarily affects the cone photoreceptors; the disease was originally mapped to a limited region of Xp11.4. We evaluated the three families from our original study with new markers and clinically reassessed all key recombinants; we determined that the critical intervals in families 2 and 3 overlapped the RP3 locus and that a status change (from affected to probably unaffected) of a key recombinant individual in family 1 also reassigned the disease locus to include RP3 as well. Mutation analysis of the entire RPGR coding region identified two different 2-nucleotide (nt) deletions in ORF15, in family 2 (delAG) and in families 1 and 3 (delGG), both of which result in a frameshift leading to altered amino acid structure and early termination. In addition, an independent individual with X-linked cone-rod dystrophy demonstrated a 1-nt insertion (insA) in ORF15. The presence of three distinct mutations associated with the same disease phenotype provides strong evidence that mutations in RPGR exon ORF15 are responsible for COD1. Genetic heterogeneity was observed in three other families, including the identification of an in-frame 12-nt deletion polymorphism in ORF15 that did not segregate with the disease in one of these families. PMID- 11857237 TI - James Sydney Stillman, MD, 1908--1998. PMID- 11857229 TI - Errata. PMID- 11857238 TI - Erratum. PMID- 11857240 TI - Richard H. Freyberg, MD, 1904--1999. PMID- 11857243 TI - Errata. PMID- 11857245 TI - Sensitivity and specificity of tests for autoantibodies: Comment on the article by Tan et al. PMID- 11857251 TI - Winners of the 1999 American College of Rheumatology Slide Competition. PMID- 11857247 TI - Arthritis & Rheumatism in 2000. PMID- 11857254 TI - James R. Klinenberg, MD, 1934--1999. PMID- 11857255 TI - Errata. PMID- 11857257 TI - Introducing the new editor of Arthritis & Rheumatism, David S. Pisetsky, MD, PHD. PMID- 11857258 TI - Joseph Lee Hollander, MD, 1910--2000. PMID- 11857259 TI - Erratum. PMID- 11857268 TI - Characterization of lipids from human brain tissues by multinuclear magnetic resonance spectroscopy. AB - Multinuclear ((1)H, (13)C, and (31)P) magnetic resonance spectroscopy are applied to the biochemical characterization of the total lipid fraction of healthy and neoplastic human brain tissues. Lipid extracts from normal brains, glioblastomas, anaplastic oligodendrogliomas, oligodendrogliomas, and meningiomas are examined. Moreover, the unknown liquid content of a cyst adjacent to a meningioma is analyzed. Two biopsies from glioblastomas are directly studied by (1)H-NMR without any treatment (ex vivo NMR). The (1)H- and (13)C-NMR analysis allows full characterization of the lipid component of the cerebral tissues. In particular, the presence of cholesteryl esters and triglycerides in the extracts of high grade tumors is correlated to the vascular proliferation degree, which is different from normal brain tissue and low grade neoplasms. The (31)P spectra show that phosphatidylcholine is the prominent phospholipid and its relative amount, which is higher in gliomas, is correlated to the low grade of differentiation of tumor cells and an altered membrane turnover. The ex vivo (1)H NMR data on the glioblastoma samples show the presence of mobile lipids that are correlated to cell necrotic phenomena. Our data allow a direct correlation between biochemical results obtained by NMR and the histopathological factors (vascular and cell proliferations, differentiation, and necrosis) that are prominent in determining brain tumor grading. PMID- 11857269 TI - UV resonance Raman study of streptavidin binding of biotin and 2-iminobiotin: comparison with avidin. AB - UV resonance Raman (UVRR) spectroscopy is used to study the binding of biotin and 2-iminobiotin by streptavidin, and the results are compared to those previously obtained from the avidin-biotin complex and new data from the avidin-2 iminobiotin complex. UVRR difference spectroscopy using 244-nm excitation reveals changes to the tyrosine (Tyr) and tryptophan (Trp) residues of both proteins upon complex formation. Avidin has four Trp and only one Tyr residue, while streptavidin has eight Trp and six Tyr residues. The spectral changes observed in streptavidin upon the addition of biotin are similar to those observed for avidin. However, the intensity enhancements observed for the streptavidin Trp Raman bands are less than those observed with avidin. The changes observed in the streptavidin Tyr bands are similar to those observed for avidin and are assigned exclusively to the binding site Tyr 43 residue. The Trp and Tyr band changes are due to the exclusion of water and addition of biotin, resulting in a more hydrophobic environment for the binding site residues. The addition of 2 iminobiotin results in spectral changes to both the streptavidin and avidin Trp bands that are very similar to those observed upon the addition of biotin in each protein. The changes to the Tyr bands are very different than those observed with the addition of biotin, and similar spectral changes are observed in both streptavidin and avidin. This is attributable to hydrogen bond changes to the binding site Tyr residue in each protein, and the similar Tyr difference features in both proteins supports the exclusive assignment of the streptavidin Tyr difference features to the binding site Tyr 43. PMID- 11857270 TI - Spectroscopic studies of interaction of chlorobenzylidine with DNA. AB - Electronic absorbance and fluorescence titrations are used to probe the interaction of chlorobenzylidine with DNA. The binding of chlorobenzylidine to DNA results in hypochromism, a small shift to a longer wavelength in the absorption spectra, and emission quenching in the fluorescence spectra. These spectral characteristics suggest that chlorobenzylidine binds to DNA by an intercalative mode. This conclusion is reinforced by fluorescence polarization measurements. Scatchard plots constructed from fluorescence titration data give a binding constant of 1.3 x 10(5) M(-1) and a binding site size of 10 base pairs. This indicates that chlorobenzylidine has a high affinity with DNA. The intercalative interaction is exothermic with a Van't Hoff enthalpy of -143 kJ/mol. This result is obtained from the temperature dependence of the binding constant. The interaction of chlorobenzylidine with DNA is affected by the pH value of the solution. The binding constant has its maximum at pH 3.0. Upon binding to DNA, the fluorescence from chlorobenzylidine is quenched efficiently by the DNA bases and the fluorescence intensity tends to be constant at high concentrations of DNA when the binding is saturated. The Stern-Volmer quenching constant obtained from the linear quenching plot is 1.6 x 10(4) M(-1) at 25 degrees C. The measurements of the fluorescence lifetime and the dependence of the quenching constant on the temperature indicate that the fluorescence quenching process is static. The fluorescence lifetime of chlorobenzylidine is 1.9 +/- 0.4 ns. PMID- 11857271 TI - Homonuclear (1)H-NMR assignment and structural characterization of human immunodeficiency virus type 1 Tat Mal protein. AB - The transacting transcriptional activator (Tat) is a viral protein essential for activation of the human immunodeficiency virus (HIV) genes, and it plays an important role in HIV induced immunodeficiency. We report the NMR structural characterization of the active Tat Mal variant that belongs to a highly virulent D-subtype HIV type-1 (HIV-1) strain (Mal) found mainly in Africa. A full Tat Mal protein (87 residues) is synthesized. This synthetic protein is active in a transactivation assay with HeLa cells infected with the HIV long terminal repeated noncoding sequences of the HIV-1 provirus (LTR) lac Z gene. Homonuclear (1)H-NMR spectra allows the sequential assignment of the Tat Mal spin systems. Simulating annealing generates 20 conformers with similar folding. The geometry of the mean structure is optimized with energy minimization to obtain a final structure. As the European variant (Tat Bru) the N-terminal region of Tat Mal constitutes the core, and there is a hydrophobic pocket composed of the conserved Trp 11 interacting with several aromatic residues. The two functional regions of Tat (basic and the cysteine-rich regions) are well exposed to the solvent. A short alpha-helix is observed in region V adjacent to the basic region. This alpha helix induces local structural variations compared to the NMR structure of Tat Bru, and it brings the cysteine-rich and basic regions closer. This study suggests that similar folding exists among Tat variants. PMID- 11857272 TI - Vibrational circular dichroism of gramicidin D in vesicles and micelles. AB - The vibrational circular dichroism (VCD) and absorption spectra of gramicidin D in three model membranes (dioctadecyldimethylammonium chloride vesicles, dimyristoyl-phosphatidylcholine vesicles, and sodium dodecyl sulfate micelles) are presented. The absorption and VCD spectra suggest that the stable gramicidin D conformation in the model membranes is different from those in organic solvents. The presence of cations does not change the membrane-bound conformation of gramicidin D. PMID- 11857273 TI - Raman and surface enhanced Raman spectroscopy of 2,2,5,5-tetramethyl-3-pyrrolin-1 yloxy-3-carboxamide labeled proteins: bovine serum albumin and cytochrome c. AB - 2,2,5,5-Tetramethyl-3-pyrrolin-1-yloxy-3-carboxamide (tempyo) labeled bovine serum albumin and cytochrome c at different pH values were prepared and investigated using Raman-resonance Raman (RR) spectroscopy and surface enhanced Raman scattering (SERS) spectroscopy. The Raman spectra of tempyo labeled proteins in the pH 6.7-11 range were compared to those of the corresponding free species. The SERS spectra were interpreted in terms of the structural changes of the tempyo labeled proteins adsorbed on the silver colloidal surface. The tempyo spin label was found to be inactive in the Raman-RR and SERS spectra of the proteins. The alpha-helix conformation was concluded to be more favorable as the SERS binding site of bovine serum albumin. In the cytochrome c the enhancement of the bands assigned to the porphyrin macrocycle stretching mode allowed the supposition of the N-adsorption onto the colloidal surface. PMID- 11857274 TI - Surface immobilization of galactose onto aliphatic biodegradable polymers for hepatocyte culture. AB - A novel surface modification method of biodegradable polymers was investigated for inducing the attachment of specific cells onto the polymer surface via ligand receptor interactions. Galactose, a targeting ligand specific to asialoglycoprotein receptors present on cell membrane of hepatocytes, was introduced on the surface of poly(D,L-lactic-co-glycolic acid) (PLGA) films. A terminal end group of carboxylic acid in PLGA was activated by dicyclohexylcarbodiimide and N-hydroxysuccinimide for the direct conjugation of lactose by reductive amination reaction. Di-block copolymers of PLGA-b poly(ethylene glycol) (PEG) having a free terminal amine group were also synthesized and used for the conjugation of galactose for the introduction of a PEG spacer between PLGA and galactose. The presence of galactose moieties on the blend film surface was characterized by measuring water contact angle and X-ray photon spectroscopy, and the amount of galactose was indirectly determined by a specific lectin-binding assay. With increasing the galactose concentration on the blend film surface, the initial attachment as well as the cell viability of hepatocyates concomitantly increased. The introduction of PEG spacer reduced the cell attachment and viability. Albumin secretion rate from hepatocytes was enhanced for galactose modified surfaces, whereas it was reduced for the surfaces not having galactose moieties. PMID- 11857275 TI - The simultaneous biosynthesis and uptake of amino acids by Lactococcus lactis studied by (13)C-labeling experiments. AB - Uniformly (13)C labeled glucose was fed to a lactic acid bacterium growing on a defined medium supplemented with all proteinogenic amino acids except glutamate. Aspartate stemming from the protein pool and from the extracellular medium was enriched with (13)C disclosing a substantial de novo biosynthesis of this amino acid simultaneous to its uptake from the growth medium and a rapid exchange flux of aspartate over the cellular membrane. Phenylalanine, alanine, and threonine were also synthesized de novo in spite of their presence in the growth medium. PMID- 11857276 TI - Numerical simulation for electrochemical cultivation of iron oxidizing bacteria. AB - A numerical simulation model was constructed for electrochemical cultivation of iron oxidizing bacterium, Thiobacillus ferrooxidans, based on Monod's dual limitation equation. In this model, two limiting factors were examined, low supply of Fe(II) ion and dissolved oxygen, from empirical viewpoints. The simulation model was constructed taking into consideration the energy balance based on the amount of the electronic flow from the electrode to bacteria via an iron ion, and then to oxygen. The model consisted of a logarithmic bacterial growth phase during the first three days, followed by a plateau and growth limitation thereafter. The predicted results were in agreement with the actual growth under electrochemical cultivation. It was predicted the growth limiting factor would be changed from insufficient supply of Fe(II) ions to that of oxygen by decreasing the value of oxygen transfer constant K, which correlated with the aeration rate. The optimum aeration rate was determined for the ideal electrochemical cultivation. The algorithm described here can be used in any electrochemical cultivation by modifying the parameters for each system. PMID- 11857277 TI - Kinetic studies on elemental sulfur oxidation by Acidithiobacillus ferrooxidans: sulfur limitation and activity of free and adsorbed bacteria. AB - The kinetics of sulfur oxidation by Acidithiobacillus ferrooxidans in shaking flasks and a 10-L reactor was studied. The observed linearity of growth and sulfur oxidation was explained by sulfur limitation. Total cell yield was not significantly different for exponential growth as compared to growth during the sulfur-limiting phase. Kinetic studies of sulfur oxidation by growing and nongrowing bacteria indicated that both free and adsorbed bacteria oxidize sulfur. Changes in the number of free bacteria rather than cells adsorbed on sulfur were better predictors of the kinetics of sulfur oxidation, indicating that the free bacteria were performing sulfur oxidation. The active growth phase always followed adsorption of bacteria on sulfur; however, the special metabolic role of adsorbed bacteria was unclear. Their activity in sulfur solubilization was considered. PMID- 11857278 TI - Efficient water removal in lipase-catalyzed esterifications using a low-boiling point azeotrope. AB - High conversions in lipase-catalyzed syntheses of esters from free acyl donors and an alcohol requires efficient removal of water preferentially at temperatures compatible to enzyme activity. Using a lipase B from Candida antarctica (CAL-B) mediated synthesis of sugar fatty-acid esters, we show that a mixture of ethyl methylketone (EMK) and hexane (best ratio: 4:1, vo/vo) allows efficient removal of water generated during esterification. Azeotropic distillation of the solvent mixture (composition: 26% EMK, 55% hexane, 19% water) takes place at 59 degrees C, which closely matches the optimum temperature reported for CAL-B. Water is then removed from the azeotrope by membrane vapor permeation. In case of glucose stearate, 93% yield was achieved after 48 h using an equimolar ratio of glucose and stearic acid. CAL-B could be reused for seven reaction cycles, with 86% residual activity after 14 d total reaction time at 59 degrees C. A decrease in fatty-acid chain length as well as increasing temperatures (75 degrees C) resulted in lower conversions. In addition, immobilization of CAL-B on a magnetic polypropylene carrier (EP 100) facilitated separation of the biocatalyst. PMID- 11857279 TI - A new fluid distribution system for scale-flexible expanded bed adsorption. AB - A new fluid distribution system designed for expanded bed adsorption was introduced and studied in a 150-cm diameter column. Based on fluid application through a rotating distributor, it eradicates the need for perforated plates, meshes, or local mixers. The effect of rotation rate on column performance was examined by fluidizing a 30-cm high bed of supports with tap water and introducing pulses of dye or acetone tracer. Linear bed expansion was seen as the superficial fluid velocity was raised from 170 x h(-1) to 450 cm x h(-1) (3000 L x h(-1) to 8000 L x h(-1)), and there was little change in expansion characteristics as distributor rotation rate was increased from 2.5 to 10 rpm. The distributor was observed to generate a flow pattern suitable for expanded bed adsorption when the supports were fluidized at a superficial fluid velocity of 283 cm center dot h(-1) and dye pulses introduced. At a rotation rate of 2.5 rpm, no significant dead zones were observed, and a discrete band was formed that moved up through the bed. Furthermore, the pattern of dye movement could be used to calculate interstitial linear fluid velocities of 460 cm x h(-1) and 572 cm x h(-1) at the column wall and center, respectively, indicating a parabolic flow profile. The distributor rotation rate giving the best operating conditions was found to be 2.5 rpm when the bed was fluidized at a flow velocity of 283 cm x h( 1) and the residence time distribution of acetone tracer examined. Under these conditions, the coefficient of axial dispersion was 6.1 x 10(-6) m(2) x s(-1) and 29 theoretical plates were measured. When the rotation rate was raised to 10 rpm, the coefficient of axial dispersion increased to 8.08 x 10(-6) m(2) x s(-1) and the number of theoretical plates decreased to 22. PMID- 11857280 TI - Characterization of a hydrogen-producing granular sludge. AB - This study demonstrated that hydrogen-producing acidogenic sludge could agglutinate into granules in a well-mixed reactor treating a synthetic sucrose containing wastewater at 26 degrees C, pH 5.5, with 6 h of hydraulic retention. A typical matured granule is 1.6 mm in diameter, 1.038 g/mL in density, 11% in ash content, and over 50 m/h in settling velocity. Treating a solution containing 12.15 g/L of sucrose at a volumetric loading rate of 48.6 g/(L x d), the reactor containing 20 g/L of granular sludge degraded 97% of sucrose. Effluent comprised 46% acetate and 49% butyrate and the methane-free biogas comprised 63% hydrogen, 35% carbon dioxide, and 2% nitrogen. Hydrogen production rate was 13.0 L/(L x d), and the yield was 0.28 L/g-sucrose. The granule had multiple cracks on the surface and comprised two morphological types of bacteria: fusiform bacilli and a spore-forming bacterium. Phylogenetic analysis showed that 69.1% of the clones were affiliated with four Clostridium species in the family Clostridiaceae, and 13.5% with Sporolactobacillus racemicus in the Bacillus/Staphylococcus group. PMID- 11857281 TI - Enzyme activation in organic solvents: co-lyophilization of subtilisin Carlsberg with methyl-beta-cyclodextrin renders an enzyme catalyst more active than the cross-linked enzyme crystals. AB - In this study we explored the efficiency of the additive methyl-beta-cyclodextrin (M beta CD) to enhance the activity and enantioselectivity of the serine protease subtilisin Carlsberg in organic solvents. These two parameters, measured for different transesterification reactions and in several solvents, are compared with results obtained by using two additional preparations of the same enzyme: lyophilized powder and cross-linked enzyme crystals (CLEC). The results suggest that co-lyophilization of subtilisin with M beta CD preserves the enzyme's active site tertiary structure rendering a highly active and enantioselective catalyst. PMID- 11857282 TI - Time-dependent solubilization of IgG in AOT-brine-isooctane microemulsions: role of cluster formation. AB - The stability and structure of protein-containing water-in-oil (w/o) microemulsions were investigated by using the large protein immunoglobulin G (IgG, MW 155,000) in a mixture comprised of brine, sulfosuccinic acid bis [2 ethylhexyl]ester (sodium salt), and isooctane. We explored factors affecting the initial uptake of IgG into the w/o microemulsion and its subsequent release to a solid (precipitate) phase, and the kinetics of the latter process. Influences of such parameters as pH, ionic strength, and protein concentration on the solubilization and precipitation of bovine IgG in the organic phase are described. The structure and dynamics in microemulsions containing bovine IgG were probed by using dynamic light scattering, and it was found that the presence of IgG in the microemulsion induced strong attractive forces between the droplets. Based on results obtained by using these various experimental approaches, a model for protein solubilization and release is proposed. In this model, we propose the formation of clusters within which bovine IgG resides and which substantially slow the kinetics of protein release from the droplets to the precipitate phase. PMID- 11857283 TI - Inline characterization of cell concentration and cell volume in agitated bioreactors using in situ microscopy: application to volume variation induced by osmotic stress. AB - A new in situ microscope (ISM) was developed and tested to perform in-line monitoring of average cell volume and cell concentration in agitated cultures subjected to osmotic stress. The ISM is directly immersed into the agitated broth in a bioreactor and generates still images of cells by using pulsed luminescent diode illumination and a virtual probe volume defined by depth of focus. This technique allows the acquisition of microscopic still images without mechanical sampling techniques. The front end of the sensor fits into a standard 25-mm port and it can be steam sterilized together with the bioreactor. The automatic image evaluation generates signals of the cell concentration and the average cell volume with a time resolution of a few minutes per data point (if a 200 MHz PC is used). Without the need for evaluation, the images can be acquired and stored at a rate of one image per 0.6 s. Hansenula anomala was cultivated as batch fermentation and monitored inline with the ISM. The ISM signal of the cell concentration agreed well with referential growth curves that were obtained from counting with a hemocytometer. The ISM signal of the average cell volume shows a gradual volume reduction as a result of the aging of the culture, and it monitors an abrupt and strong cell contraction if osmotic shocks are generated in the bioreactor. Systematic in vitro studies of osmotic shocks were performed by applying the ISM to agitated culture samples of H. anomala. The volume signal of H. anomala during osmotic shocks showed a very fast cell contraction within less than a second. Within half an hour after the shocks, no signal drifts were observed, which would indicate volume restoration. These findings suggest that the ISM volume signal can be used as an inline indicator of osmotic stress in cell cultures. PMID- 11857284 TI - Enhancement of shikonin production in single- and two-phase suspension cultures of Lithospermum erythrorhizon cells using low-energy ultrasound. AB - This work demonstrates the use of low-energy ultrasound (US) to enhance secondary metabolite production in plant cell cultures. Suspension culture of Lithospermum erythrorhizon cells was exposed to low-power US (power density < or = 113.9 mW/cm(3)) for short periods (1-8 min). The US exposure significantly stimulated the shikonin biosynthesis of the cells, and at certain US doses, increased the volumetric shikonin yield by about 60%-70%. Meanwhile, the shikonin excreted from the cells was increased from 20% to 65%-70%, due partially to an increase in the cell membrane permeability by sonication. With combined use of US treatment and in situ product extraction by an organic solvent, or the two-phase culture, the volumetric shikonin yield was increased more than two- to threefold. Increasing in the number of US exposures during the culture process usually resulted in negative effects on shikonin yield but slight stimulation of shikonin excretion. US at relatively high energy levels caused slight cell growth depression (maximum 9% decrease in dry cell weight). Two key enzymes for the secondary metabolite biosynthesis of cells, phenylalanine ammonia lyase and p-hydroxybenzoic acid geranyltransferase, were found to be stimulated by the US. The US stimulation of secondary metabolite biosynthesis was attributed to the metabolic activity of cells activated by US, and more specifically, the defense responses of plant cells to the mechanical stress of US irradiation. PMID- 11857285 TI - Quantitative analyses of anaerobic wastewater treatment processes: identifiability and parameter estimation. AB - We investigated the problem of identifying the parameters of a nonlinear fifth order model describing the population dynamics of two main bacterial groups in an anaerobic wastewater treatment process. In addition to addressing problems concerning structural and practical identifiability, we also analyzed how mathematical descriptions of bacterial population dynamics can model real data. Using three data sets recorded under different experimental conditions, we estimated important biochemical parameters and demonstrated that our model could describe the data successfully. Parameters, which are simultaneously determined using information from all three experiments, have more reliable estimates. We conclude that, after appropriate estimation, this model can be used for optimization and the control of continuous processes. PMID- 11857286 TI - Asymmetric sulfoxidations mediated by alpha-chymotrypsin. AB - The oxidation of aryl alkyl sulfides with H(2)O(2) in aqueous solution is a reasonably facile reaction producing racemic sulfoxides. We show that in the presence of the hydrolytic enzyme alpha-chymotrypsin such a sulfoxidation is accelerated and, more importantly, becomes stereoselective. With phenyl isobutyl sulfide as a model, the chymotrypsin-mediated, highly asymmetric oxidation is shown to occur in the hydrophobic binding pocket of the enzyme active site. The stereoselectivity of the chymotrypsin-mediated sulfoxidations is correctly explained by means of structure-based molecular modeling of the enzyme-sulfide complexes. PMID- 11857287 TI - A biosensor for the detection of triazine and phenylurea herbicides designed using Photosystem II coupled to a screen-printed electrode. AB - A biosensor for the detection of triazine- and phenylurea-type herbicides was constructed using isolated Photosystem II (PS II) complexes as a biosensing element. PSII isolated from the thermophilic cyanobacterium Synechococcus elongatus was immobilized on the surface of a screen-printed sensor composed of a graphite working electrode and Ag/AgCl reference electrode deposited on a polymeric substrate. The biosensor was mounted in a flow microcell with illumination. The principle of the detection was based on the fact that herbicides selectively block PSII electron transport activity in a concentration dependent manner. Changes of the activity were registered amperometrically as the rate of photoreduction of an artificial electron acceptor. The setup resulted in a reusable herbicide biosensor with a good stability (half-life of 24 h) and limit of detection of approximately 10(-9) M for diuron, atrazine and simazine. PMID- 11857288 TI - Cisplatin, fludarabine, and cytarabine: a novel, pharmacologically designed salvage therapy for patients with refractory, histologically aggressive or mantle cell non-Hodgkin's lymphoma. AB - BACKGROUND: Based on in vitro synergism, the combination of cytarabine (ara-C) and cisplatin is the basis of many salvage regimens for patients with aggressive non-Hodgkin lymphoma (NHL). However, patients with previously refractory disease are significantly less likely to respond, stimulating the search for novel salvage regimens. In vitro, fludarabine enhances the cytotoxicity of both ara-C and cisplatin, increasing ara-C incorporation into DNA and inhibiting repair of platinum/DNA adducts, suggesting that the combination of cisplatin, fludarabine, and ara-C (PFA) may have clinical utility. METHODS: A Phase-II study of a 96 hour continuous infusion of cisplatin with two timed-sequential couplets of fludarabine and ara-C together with granulocyte colony stimulating factor was performed in 45 patients with previously refractory, histologically aggressive or mantle cell NHL. RESULTS: Patients had predominantly diffuse large cell and/or immunoblastic NHL or its variants (80%), or they had mantle cell lymphoma (18%). Overall, 93% of patients had previously refractory disease, with a median International Prognostic Index score of 3. A median of 2 cycles per patient were delivered (range, 1-4 cycles) with significant myelosuppression; there were medians of 2 days of neutropenia < 0.5 x 10(9)/L (range, 0-12 days) and 3 days of thrombocytopenia < 20 x 10(9)/L (range, 0-24 days). This was more severe in older patients and was cumulative with successive cycles. Thirty-five percent of cycles were complicated by infections, nausea and emesis were prominent, but other nonhematologic toxicity was mild. Peripheral blood progenitor cells were mobilized adequately after the first cycle, but collections were impaired after more prolonged therapy. The overall response rate was 48% (7% of patients had complete responses, and 41% of patients had partial responses), with one toxic death due to tumor-lysis syndrome. Patients with mantle cell lymphoma were more likely to respond than patients with other histologies (88% vs. 39%, respectively; P = 0.019), although three of eight patients had relapsed rather than refractory disease. The median remission duration was 4 months, with 28% of potentially eligible patients able to proceed to subsequent high dose therapy. The actuarial 2 year survival rates were 20% +/- 6% overall and 50 +/- 18% for patients with mantle cell lymphoma. CONCLUSIONS: Given the adverse outlook for these patients, the results are promising, particularly for patients with mantle cell lymphoma, and suggest that the addition of fludarabine as a potential biochemical modulator may enhance the activity of cisplatin and ara-C. This is associated with significant cumulative (but manageable) myelosuppression. This paradigm, in which a nucleoside analogue is used to inhibit the repair of platinum/DNA adducts, also may be applicable for the treatment of patients with other types of platinum-sensitive tumors. PMID- 11857289 TI - Irinotecan hydrochloride for the treatment of recurrent and refractory non Hodgkin lymphoma: a single institution experience. AB - BACKGROUND: Irinotecan hydrochloride (CPT-11) has a broad range of antitumor activity and has demonstrated little cross-resistance with doxorubicin or vincristine. In the current study, the authors investigated the efficacy and adverse effects of irinotecan in the treatment of recurrent and refractory non Hodgkin lymphoma, for which current therapies appear to be unsatisfactory. METHODS: Irinotecan was administered by intravenous infusion at a dose of 40 mg/m(2)/day for 3 days, and this regimen was repeated 2-3 times at weekly intervals, followed by 2 weeks off therapy. The subjects were 48 patients with recurrent or refractory non-Hodgkin lymphoma. The histologic classification (Working Formulation) was low grade in 8 patients, intermediate grade in 36 patients, high grade in 1 patient, and other (angiocentric lymphoma, Ki-1 lymphoma, and unidentified) in 3 patients. RESULTS: Forty-five patients were determined to be evaluable. Therapy resulted in a complete disease remission in 2 patients and a partial remission in 15 patients. The response rate was 37.8%. The median duration of response was 64 days and the median time to disease progression was 77 days. The median survival time was 422 days. Major adverse reactions included myelosuppression and gastrointestinal toxicity. Leukopenia, anemia, and thrombocytopenia of Grade 3 or 4 (according to the National Cancer Institute Common Toxicity Criteria) was observed in 63.0%, 30.4%, and 6.5% of the patients, respectively, and Grade 3 or 4 diarrhea occurred in 30.4% of patients. Treatment was withdrawn because of diarrhea in three patients. Because of myelosuppression and diarrhea, approximately 67% of the patients required changes to the regimen, including dose reduction, prolongation of the interval between treatments, and reducing the number of days of consecutive treatment. CONCLUSIONS: The results of the current study suggest the activity of irinotecan as salvage therapy for patients with recurrent and refractory non-Hodgkin lymphoma. However, the optimum dosing schedule remains to be determined. PMID- 11857290 TI - Results with chemotherapy comprised of cyclophosphamide, doxorubicin, vincristine, and prednisone followed by radiotherapy with or without prechemotherapy surgical debulking for patients with bulky, aggressive lymphoma. AB - BACKGROUND: The authors performed a case-control analysis of local control, progression free survival, and overall survival in patients with Stage I-II aggressive lymphomas measuring > or = 7 cm in greatest dimension who were treated initially with or without surgical debulking: All patients then received cyclophosphamide, doxorubicin, vincristine, and prednisone (CHOP) based chemotherapy followed by involved field radiotherapy. METHODS: From May 1975 through May 1996, 50 patients were treated with (n = 25 patients) or without (n = 25 patients) resection of > 80% of their bulky lymphomas. Chemotherapy consisted of 3-12 cycles of CHOP. In general, patients who underwent debulking received three cycles of chemotherapy, whereas patients who did not undergo debulking received at least six cycles of chemotherapy. The total radiotherapy dose was 40.8 grays (Gy) +/- 4.2 Gy (mean +/- standard deviation). RESULTS: The median follow-up was 62 months. Patients who underwent debulking were similar prognostically to patients who did not. There was a trend toward improved local control (5 year rates: 96% vs. 80%; P = 0.10) and overall survival (5 year rates: 83% vs. 71%; P = 0.18) in patients who underwent debulking compared with patients who did not, respectively. Progression free survival was significantly better for patients who underwent debulking compared with patients who did not (5 year rates: 88% vs. 62%, respectively; P = 0.04). CONCLUSIONS: Because this study was retrospective, selection bias may account for the observed difference in progression free survival. Because it is unlikely that a trial randomizing patients with bulky, aggressive lymphoma to surgery will be conducted, the authors' current efforts are focused on escalation of the total radiotherapy dose as a possibly less costly and less morbid approach toward improving progression free survival for these patients. PMID- 11857291 TI - Breast carcinoma in women age 25 years or less. AB - BACKGROUND: Although several studies have examined breast carcinoma in young women aged < or = 35 years at diagnosis, there are only occasional cases reported in very young women aged < or = 25 years, and, to the authors' knowledge, no series are available. The presentation, tumor biology, behavior, and outcome of breast carcinoma in very young women are not known, and the rarity of breast malignancy within this age group could lead to diagnostic delays. METHODS: The tumor characteristics and survival of 15 women aged < or = 25 years at the time of diagnosis, have been reviewed and compared with women aged 26-35 years under the care of Guy's Hospital's Breast Unit during the same period of time. Where appropriate, the two groups were individually matched for tumor size (clinical measurement) and histologic grade. RESULTS: Fifteen cases were examined, with a median follow-up of 108 months and a median age of 24 years. The median duration of symptoms was 4 weeks, and the median tumor size was 20 mm. Two patients had ductal carcinoma in situ (DCIS) only, while the other 13 patients had invasive carcinomas, none of which were Grade I. A mastectomy was performed on 8 out of 15 patients (53%). Axillary nodal metastases were present in 4 out of 12 patients (33%). Of the 13 cases of invasive disease, 9 out of 13 patients (69%) experienced recurrence and died of breast carcinoma. Median disease free survival for patients with invasive disease was 86 months. There was no difference in overall survival between the patients aged < or = 25 years and those aged 26-35, but taken together young women < or = 35 had a worse prognosis than women between 36 and 65, due to a higher incidence of high grade and estrogen receptor negative tumors. CONCLUSIONS: The current study suggests that among young women with breast carcinoma there is no difference in prognosis between the very young and the young. Despite two thirds of patients being node negative, the high mortality rate indicates a need for an optimal selection of adjuvant therapy among these cases. PMID- 11857292 TI - Functionally active estrogen receptor isoform profiles in the breast tumors of African American women are different from the profiles in breast tumors of Caucasian women. AB - BACKGROUND: Several cancer surveys have shown that African-American women (AAW) develop highly aggressive breast tumors and experience about three times higher mortality rates compared with other populations. Generally, breast tumors in AAW are poorly differentiated or undifferentiated and exhibit increased frequency of nuclear atypia, higher mitotic activity, higher S-phase fraction, and tumor necrosis. The molecular factors responsible for these tumor characteristics are mostly unknown. METHODS: To explore whether the aggressive tumor biology observed in AAW is related to distinct alterations in estrogen receptor (ER) isoforms, the relative expression levels of four functionally active ER isoform mRNAs, ERalpha wild type, ERbeta wild type, ERalpha exon 3delta, and ERalpha exon 5delta, were measured by reverse transcriptase-polymerase chain reaction analysis in 18 immunohistochemically ERalpha positive tumors and in 6 ERalpha negative tumors and their matched normal tissues. RESULTS: In the tumors of AAW, the protective ERbeta isoform was decreased significantly compared with matched normal tissues (paired t test; n = 24 patients; P = 0.0018). In addition, both the constitutively active ERalpha exon 5delta and the dominant negative ERalpha exon 3delta mRNA levels were elevated in tumor tissues compared with matched normal tissues (paired t tests; n = 24 patients; P = 0.0002 and P = 0.024, respectively). CONCLUSIONS: The data presented here show for the first time that functionally active ER isoform profiles in the breast tumors of AAW are different from those in Caucasian women. The tumors in AAW are characterized by decreased levels of the protective ERbeta isoform and elevated levels of the constitutively active ERalpha exon 5delta isoform. Variations in estrogen-mediated signaling because of the alterations in these two ER isoforms may account in part for differences in tumor biology between AAW and Caucasian women. PMID- 11857293 TI - The World Health Organization histologic classification system reflects the oncologic behavior of thymoma: a clinical study of 273 patients. AB - BACKGROUND: Although the histologic classification of thymic epithelial tumors has been confusing and controversial, an agreement on the universal classification system for thymic epithelial tumors was achieved by the World Health Organization (WHO) in 1999. The authors previously reported that the WHO histologic classification system reflects invasiveness and immunologic function of thymic epithelial tumors. In this subsequent study, they examined the prognostic significance of this classification system. METHODS: Clinical features as well as postoperative survival of patients with thymoma, but not thymic carcinoma, were examined with reference to WHO histologic classification based on an experience with 273 patients over a 44-year period. RESULTS: There were 18 type A tumors, 77 type AB tumors, 55 type B1 tumors, 97 type B2 tumors, and 26 type B3 tumors. In patients with type A, AB, B1, B2, and B3 tumors, the respective proportions of invasive tumor were 11.1%, 41.6%, 47.3%, 69.1%, and 84.6%; the respective proportions of tumors with involvement of the great vessels were 0%, 3.9%, 7.3%, 17.5%, and 19.2%; and the respective 20-year survival rates were 100%, 87%, 91%, 59%, and 36%. According to the Masaoka staging system, the 20-year survival rates were 89%, 91%, 49%, 0%, and 0% in patients with Stage I, II, III, IVa, and IVb disease, respectively. By multivariate analysis, the Masaoka staging system and the WHO histologic classification system were significant independent prognostic factors, whereas age, gender, association with myasthenia gravis, completeness of resection, or involvement of the great vessels were not significant independent prognostic factors. CONCLUSIONS: This study showed that histologic appearance reflects the oncologic behavior of thymoma when the WHO classification system is adopted. The WHO classification system may be helpful in clinical practice for the assessment and treatment of patients with thymoma. PMID- 11857294 TI - Functional imaging of Langerhans cell histiocytosis by (111)In-DTPA-D-Phe(1) octreotide scintigraphy. AB - BACKGROUND: Langerhans cell histiocytosis is a rare group of diseases of unknown pathogenesis, variable clinical presentation, and behavior. It encompasses highly aggressive, often fatal, and indolent diseases, with a variety of intermediate forms between the two extremes. Localization and monitoring of Langerhans cell histiocytosis deposits are based on the combined use of different imaging modalities, which include magnetic resonance, computed tomography, and nuclear medicine procedures. Somatostatin receptor scintigraphy recently has been used to image immune-mediated diseases with excellent results. METHODS: In this study, the authors have evaluated the potential role of somatostatin receptor scintigraphy by using (111)In-DTPA-D-Phe(1)-octreotide in a series of patients with Langerhans cell histiocytosis. Eight consecutive patients (2 males, 6 females; mean age, 43 years; age range, 29-60 years) with histologically proven Langerhans cell histiocytosis, in various phases of their disease, were imaged 24 hours after intravenous injection of 111-222 MBq of (111)In-DTPA-D-Phe(1) octreotide, by using total body and spot view images. The objective of the study was to preliminarily evaluate this technique in the evaluation of Langerhans cell histiocytosis. RESULTS: Most Langerhans cell histiocytosis deposits were clearly documented by somatostatin receptor scintigraphy, independently from the anatomic location (i.e., skeleton, mucosae, soft tissue, etc.). The correlation between conventional imaging modalities and somatostatin receptor scintigraphy results was fairly good, though this was not considered a primary objective of the study. CONCLUSIONS: Among the various imaging modalities for Langerhans cell histiocytosis, somatostatin receptor scintigraphy is unique, because it accurately depicts the active sites of disease and thus can help in monitoring the response to treatment, together with conventional imaging modalities. Further studies are necessary to confirm the diagnostic value of this technique and clarify the biologic significance of the expression of somatostatin receptors in Langerhans cell histiocytosis. PMID- 11857295 TI - CPT-11 plus cisplatin in patients with advanced, untreated gastric or gastroesophageal junction carcinoma: results of a phase II study. AB - BACKGROUND: This Phase II study assessed the response rate and toxicity profile of the combination CPT-11 and cisplatin administered weekly to patients with untreated, advanced adenocarcinoma of the stomach or the gastroesophageal junction. METHODS: Patients with histologic proof of adenocarcinoma of the stomach or the gastroesophageal junction with adequate liver, kidney, and bone marrow functions were treated with 65 mg/m(2) CPT-11 plus 30 mg/m(2) cisplatin, both administered intravenously 1 day per week for 4 consecutive weeks, followed by a recovery period of 2 consecutive weeks. The response rate, time to disease progression, survival, and toxic effects were analyzed. RESULTS: Thirty-six of 38 registered patients (95%) were assessable. The median number of 6-week cycles per patient was 2.5 (range, 1-7 6-week cycles). Four patients (11%) achieved a complete response, and 17 patients (47%) had a partial response for an overall response rate of 58%. The median time to progression of carcinoma was 24 weeks, and the median survival was 9 months (range, 1-23+ months). There was one treatment-related death. Major toxic effects included diarrhea, neutropenia, and fatigue. Ninety percent of all planned doses were delivered on time; however, 53 of 79 canceled or delayed weekly doses (66%) occurred in the third or fourth week of the therapy cycle. CONCLUSIONS: The combination of CPT-11 and cisplatin is active against gastric or gastroesophageal adenocarcinoma and needs to be studied further. A modification in doses and schedules may be warranted to make the regimen more tolerable to patients. The addition of other active drugs or radiation therapy to this regimen would be of interest. PMID- 11857296 TI - Relevance of biologic markers in colorectal carcinoma: a comparative study of a broad panel. AB - BACKGROUND: Although pathologic stage is currently the main prognostic indicator for patients with colorectal carcinoma (CRC), mounting evidence suggests that, in its current form, it is insufficient to predict clinical outcome. To assess biologic markers of primary CRC that may improve clinical staging and provide useful information for the application of novel therapeutic strategies, the authors investigated a panel of markers that included transforming growth factor alpha (TGF alpha), epithelial growth factor receptor (EGF-R; the protein product of the c-erb B2/HER-2 oncogene), matrix metalloproteinase 2 (MMP-2), insulin-like growth factor II (IGF-II), vascular endothelial growth factor (VEGF), and angiogenesis, as evaluated by microvessel density (MVD). METHODS: Two groups of CRC were studied: 1) surgical samples from patients who achieved a disease free survival of at least 6 years (CRC-M0) and 2) surgical specimens of both primary tumors and synchronous or metachronous liver metastases (CRC-M1). RESULTS: Chi square analysis revealed that expression levels of TGF-alpha, c-erb B2/HER-2, MMP 2, IGF-II, VEGF, and MVD (but not EGF-R) were significantly higher in CRC-M1 samples compared with CRC-M0 samples (P < 0.001, P < 0.05, P < 0.001, P < 0.001, P < 0.01, and P < 0.001, respectively). Logistic regression analysis showed that TGF-alpha, IGF-II, and MMP-2 had significantly greater expression in CRC-M1 samples independent of the other variables (including tumor classification, histologic grade, and patient age). If all three markers had > or = 25% expression, then the probability of developing liver metastasis was 99.5%. CONCLUSIONS: Based on the evidence of this study, TGF-alpha MMP-2, and IGF-II seem suitable candidates for a selective panel of markers designed to provide significant additional information with respect to the current pathologic staging system for patients with colorectal carcinoma. PMID- 11857297 TI - Prognostic importance of tumor size for localized conventional (clear cell) renal cell carcinoma: assessment of TNM T1 and T2 tumor categories and comparison with other prognostic parameters. AB - BACKGROUND: The T1 and T2 classifications of the International Union Against Cancer TNM classification system for renal cell carcinoma are based on primary tumor size, and in various editions of the classification, the cut points between T1 and T2 have been amended to provide clinical utility. In the current edition, the T1/T2 cut point is less than or equal to and greater than 7 cm. and more recently a subdivision of the T1 classification (less than or equal to and < 4 cm) has been proposed to identify patients suitable for partial nephrectomy. This study investigates the prognostic significance of tumor size in a series of organ confined clear cell renal cell carcinomas. METHODS: One hundred thirty cases of organ-confined clear cell renal cell carcinomas, with a minimum of 5 years' follow-up, were identified from the New South Wales Cancer Registry. Tumor size was compared with survival using the method of Kaplan and Meier for TNM size categories, and proportional hazards regression was used for assessing size as a continuous variable. Proportional hazards regression also was used for multivariable comparisons of size and other prognostic parameters (Fuhrman grade, AgNOR score, and Ki-67 index) against survival. RESULTS: Of 116 cases for which tumor dimension was recorded, 25 patients had died of cancer-related causes. Primary tumor size ranged from 12 to 140 mm (mean, 57.3 mm). The association between survival and size was significant irrespective of the TNM classification and was also significant when size was modeled continuously (P = 0.000125, hazard of death increased by 3.51 times for each doubling of tumor size). On univariate analysis, Fuhrman grade (P = 0.04) and AgNOR score (P = 0.015) were associated with survival; however, on multivariate analysis only tumor size retained significance. CONCLUSIONS: Although the cut point of T1 and T2 TNM categories and the proposed T1 subdivision cut point correlate with survival, our finding that size is a continuous variable indicates that as a prognostic parameter for clear cell renal cell carcinoma, primary tumor size is relative rather than indicative. PMID- 11857298 TI - Higher doses of mitoxantrone among men with hormone-refractory prostate carcinoma: a Cancer and Leukemia Group B study. AB - BACKGROUND: Mitoxantrone in combination with a low-dose glucocorticoid has been shown to produce more favorable outcomes among men with hormone-refractory prostate carcinoma than glucocorticoid alone. Therefore, the authors sought to determine the safety and activity of higher doses of mitoxantrone in combination with granulocyte-macrophage colony-stimulating factor (GM-CSF) and a glucocorticoid in preparation for a possible Phase III trial comparing standard to dose-escalated mitoxantrone. METHODS: This Phase II trial enrolled 45 patients from October 1996 to March 1998. Twenty-one patients without pelvic irradiation (Arm I) received 21 mg/m(2) of mitoxantrone every 3 weeks, and 24 patients who had received pelvic irradiation (Arm II) were given 17 mg/m(2) on the same schedule. All patients received 40 mg of hydrocortisone in divided doses daily and GM-CSF at 500 microg/daily for a minimum of 10 days per cycle beginning on the third day of the cycle. RESULTS: In Arm I, 33% of assessable patients achieved a partial response, 50% had a > or = 50% decline in their PSA, and 35% had a > or = 75% decline in PSA values. The comparable numbers in Arm II were 24%, 48%, and 35%, respectively. The median survival times were 12 months in Arm I and 14 months in Arm II. Treatment had to be discontinued in 13% of patients because of thrombocytopenia. No other significant toxicities were encountered. CONCLUSIONS: Higher doses of mitoxantrone (17 and 21 mg/m(2)) were associated with activity comparable to many estramustine combinations and generally were well tolerated. However, because the degree and frequency of thrombocytopenia were greater than that observed with standard dose mitoxantrone (12-14 mg/m(2)), and because the median survival is apparently comparable to standard dose mitoxantrone, this approach to HRPC cannot be recommended for Phase III testing. PMID- 11857299 TI - Sialosyl-Le(x) expression defines invasive and metastatic properties of bladder carcinoma. AB - BACKGROUND: Two types of transitional bladder carcinoma have been distinguished based on macroscopic morphology: type A papillary carcinomas, with papillomatous surface outgrowth without infiltration into muscular layer, and type B nodular carcinomas, with a nonpapillomatous surface appearance, most of which display infiltrative growth through muscular layer, and some of which display lymphatic or blood-borne metastasis. However, there is no specific predictor at early stages for later invasive and metastatic clinical outcome of patients with type B tumors. METHODS: The study included 1) glycosphingolipid (GSL) composition of type A and B tumors; 2) histologic and immunohistologic patterns of nodular (type B) bladder carcinoma from 44 patients based on a special sampling procedure termed whole-layer core biopsy (WLCB) using the antisialosyl-Le(x) (anti-SLe(x); SLe(x): NeuAcalpha3Galbeta4[Fucalpha3]GlcNAcbeta3Galbeta4GlcCer) SNH3 antibody or other antibodies; 3) comparison of the incidence of metastasis in patients with SNH3 positive versus SNH3 negative primary tumors and of 5-year survival curves; 4) comparison of bladder carcinoma cell lines from tumors with high versus low malignancy in terms of expression patterns of SLe(x), SLe(a), and other carbohydrates, E-selectin dependent adhesion, and transcript levels of five fucosyltransferases. RESULTS: Anti-SLe(x) monoclonal antibody (mAb) SNH3 staining of WLCB samples from 44 type B tumors showed that the majority of tumors (n = 31 patients) were SNH3 positive and the minority (n = 13 patients) were SNH3 negative. SNH3 positive patients had more lymph node or blood-borne metastasis and lower 5-year and 7-year survival rates, as indicated by Kaplan-Meier curves (P = 0.001). Staining of samples with other antibodies, including FH6 and CA19-9, was not correlated with long-term survival. Determination of GSL composition in extracts showed that SLe(x) ganglioside was present in all three patients with nodular tumors but absent in all three patients with papillary tumors tested. Bladder carcinoma cell lines from invasive tumors that maintained their metastatic properties were SNH3 positive, showed high levels of alpha1,3 fucosyltransferase VI (FT-VI) and FT-VII, and displayed E-selectin dependent adhesion. Cell lines from noninvasive tumors or normal bladder epithelia were negative for SNH3 reactivity, FT-VI, and FT-VII, and E-selectin dependent adhesion. CONCLUSIONS: SLe(x) expression in primary bladder carcinoma, defined by the mAb SNH3, is a predictor of invasive and metastatic outcome. No other carbohydrate epitope examined to date has equal prognostic value. PMID- 11857300 TI - Rapid rise of serum prostate specific antigen levels after discontinuation of the herbal therapy PC-SPES in patients with advanced prostate carcinoma: report of four cases. AB - BACKGROUND: PC-SPES is an herbal supplement whose mechanisms of action are poorly understood, but may be estrogenic. The objective of the current report is to describe the effects of discontinuing PC-SPES treatment in four patients with androgen-independent prostate carcinoma. METHODS: Patient charts were retrospectively reviewed. A MEDLINE search was performed to investigate whether these effects of PC-SPES had been previously reported. RESULTS: Four men whose metastatic prostate carcinoma progressed despite androgen ablation and subsequent PC-SPES treatment are described. All four patients developed a rapid increase in serum prostate specific antigen (PSA) within one month of stopping PC-SPES, ranging from 345% to 880%. Two patients increased their PSA levels to 1300% and 1400% after 7 weeks. Compared to the rate of rise of PSA levels prior to and during PC-SPES therapy, the rise after stopping this treatment was much higher than expected. Clinical symptoms remained relatively stable despite the serologic changes. CONCLUSIONS: Discontinuing PC-SPES therapy can be associated with a rapid rise in PSA. To the authors' knowledge, this effect has not been reported previously. This effect should be considered in the design of clinical trials as well as in the standard management of androgen-independent prostate carcinoma patients. PMID- 11857301 TI - Differential expression of the mismatch repair gene hMSH2 in malignant prostate tissue is associated with cancer recurrence. AB - BACKGROUND: Mismatch repair (MMR) genes are responsible for coordinated correction of misincorporated nucleotides formed during DNA replication. Inactivating mutations in MMR genes have been described in sporadic cancers and a hereditary cancer predisposition syndrome. Mismatch repair deficiency causes instability at microsatellites and increased mutation rates. Although microsatellite instability (MSI) has been described in high-grade and lymph node positive prostate carcinoma specimens, an analysis comparing hMSH2 expression, MSI, and outcome in clinically organ confined prostate carcinoma has not been reported. METHODS: Immunohistochemical analysis of benign and malignant prostate tissue from 101 patients was performed using a monoclonal antibody specific for the hMSH2 protein. Expression was correlated with MSI using dinucleotide repeat markers and laser-captured microdissected DNA from normal and tumor cells. hMSH2 protein expression and MSI were assessed with respect to pathologic stage, Gleason score, and time to detectable serum prostate specific antigen (PSA) after prostatectomy in patients with clinically localized prostate carcinoma. RESULTS: In normal glands, hMSH2 staining was minimal to low and confined to the basal cell layer. In 32% of benign prostatic hyperplasia cases, hMSH2 staining was increased in the basal and luminal cell layers whereas 71% of cancer specimens had uniform moderate to high staining. Microsatellite instability was detected in 60% of absent to low staining and 26% of moderate to high staining prostate carcinoma specimens. Differential staining in benign versus malignant prostate tissues was statistically significant (P < 0.001) as was the correlation between absent to low hMSH2 staining and presence of MSI (P = 0.028). Decreased risk for PSA recurrence after radical prostatectomy correlated with absent to low hMSH2 staining in malignant prostate tissue but was only marginally significant (P = 0.05 for 24 month recurrence and P = 0.08 for overall time to PSA recurrence). CONCLUSIONS: The results of the current study demonstrate differential hMSH2 expression in benign and malignant prostate tissue. Moreover, hMSH2 expression is altered in a subset of clinically localized prostate carcinoma specimens independent of pathologic stage and Gleason pattern. A statistically significant correlation between hMSH2 immunohistochemical staining intensity and MSI also was identified in prostate carcinoma specimens. Furthermore, the time to cancer recurrence as determined by detectable serum PSA after prostatectomy was associated with hMSH2 staining intensity. Taken together, our results suggest that hMSH2 gene expression in prostate carcinoma may be a useful prognostic marker for outcome in men with clinically organ confined prostate carcinoma. PMID- 11857302 TI - Clinical usefulness of color Doppler ultrasound in patients with endometrial hyperplasia and carcinoma. AB - BACKGROUND: The objective of this study was to examine the usefulness of transvaginal color Doppler ultrasound (TV-CDU) in differentiating between endometrial hyperplasia (EH) and endometrial carcinoma (EC) and in predicting tumor spread in patients with EC. METHODS: Seventy-one postmenopausal patients were enrolled with either EH or EC that had been diagnosed by endometrial biopsy. The presence or absence of intratumoral blood flow was assessed by TV-CDU. The intratumoral blood flow characteristics were analyzed using the resistance index (RI), pulsatility index (PI), and peak systolic velocity (PSV). The endometrial thickness also was measured in all patients by gray-scale sonography. The correlation of these sonographic findings with histologic type, tumor grade, surgical stage, myometrial invasion, or the presence or absence of pelvic lymph node metastasis was then evaluated in patients with EC. RESULTS: Although there were no patients with EC with endometrial thickness measuring < 5 mm, no significant difference was found in the mean value of endometrial thickness between patients with EH (n = 18 patients; 16.2 mm +/- 15.9 mm) and patients with EC (n = 53 patients; 18.7 mm +/- 17.1 mm). Intratumoral blood flow was detected in significant numbers of patients who had EC (71.7%; 38 of 53 patients) compared with patients who had EH (5.6%; 1 of 18 patients; P < 0.0001). Thus, no patients with EH showed any blood flow in the endometrial lesions, except for one patient who had EH complicated by pyometra. In patients with EC, the positive rate of intratumoral blood flow was correlated significantly with myometrial invasion, tumor grade, and pelvic lymph node metastasis (P < 0.05; Cochran-Armitage trend test). No associations were found between RI, PI, or PSV and the clinicopathologic parameters examined, including surgical stage. CONCLUSIONS: TV CDU may be more useful in differentiating between EH and EC than measuring endometrial thickness by transvaginal gray-scale sonography. For patients with EC, the detection of intratumoral blood flow may be helpful in distinguishing between low-grade and high-grade tumors and predicting myometrial invasion. However, intratumoral blood flow analysis using RI, PI, or PSV may not be useful for predicting tumor spread before surgery. PMID- 11857303 TI - Circulating tumor cells in the peripheral blood and bone marrow of patients with ovarian carcinoma do not predict prognosis. AB - BACKGROUND: Ovarian carcinoma is apparently restricted for a long time to the peritoneal cavity. However, about 50% of patients with a surgically documented complete intraabdominal response experience later recurrence. Occult hematogenous micrometastases are common to most epithelial malignancies and have recently been found in 30% of bone marrow samples of ovarian carcinoma patients, as examined by immunocytochemistry. Moreover, these findings were associated with poor progression-free and overall survival. The aim of the current study was to evaluate the possible prognostic significance of tumor cells detected in the peripheral blood and bone marrow of ovarian carcinoma patients by an immunomagnetic method. METHODS: In a total of 90 patients with histologically proven epithelial ovarian carcinoma, blood and (in 73 cases) bone marrow samples were taken. Tumor cells were identified by a microbead coated with the antibody MOC-31, which recognizes an epitope regularly expressed on ovarian carcinoma cells. RESULTS: The authors detected carcinoma cells in the bone marrow in 21% of ovarian carcinoma patients, and in the peripheral blood in 12% of patients. Mean overall survival was 25 and 28 months for patients with or without circulating tumor cells, respectively. CONCLUSIONS: Ovarian carcinoma cells seem to reach peripheral circulation more frequently than expected. However, in contrast to an earlier report, detection of tumor cells in the bone marrow and/or blood was not associated with poor prognosis in ovarian carcinoma patients. This discrepancy remains unexplained, but characterization of circulating ovarian carcinoma cells for their malignant and metastatic capacity is clearly warranted. PMID- 11857304 TI - p53 and Ki-67 as markers of radioresistance in head and neck carcinoma. AB - BACKGROUND: p53 and Ki-67 are regarded as potential interesting predictors of radioresistance, although their exact influence awaits confirmation on a large cohort of uniformly treated patients. METHODS: In a retrospective cohort of 304 patients with squamous cell carcinoma of the head and neck who were treated with radical radiotherapy, the expression levels of p53 and Ki-67 were assessed by immunohistochemistry. Local control and survival curves were generated for p53 and Ki-67 using the Kaplan-Meier method. The difference between curves was calculated in univariate and multivariate analyses. RESULTS: The overexpression of p53 was associated with local treatment failure (P = 0.01) but not with survival (P = 0.09). In a Cox analysis, p53 overexpression remained an independent predictor of local failure, with a relative risk of local failure of 1.5 (P = 0.05). Low proliferation (Ki-67 < 20%) was a significant factor in local failure for patients with tumors of the oral cavity only (P = 0.01). Patients with both unfavorable immunohistochemical markers (p53 overexpression and low proliferation) had a 45% rate of local control compared with a 67% rate for all other combinations (P = 0.002). This association was even more significant in patients with T1-T2 lesions (45% vs. 77%; P = 0.0002). CONCLUSIONS: The results support the role of p53 as an independent predictor of local failure in patients with squamous cell carcinoma of the head and neck who are treated by radical radiotherapy, suggesting that it may predict radioresistance. Combined with p53, Ki-67 may help in the better selection of patients for radiotherapy, especially for patients with early-stage tumors. Prospective studies are now needed to confirm these results and to define better the role of these markers in the management of patients with head and neck carcinoma. PMID- 11857305 TI - Detection of cell free Epstein-Barr virus DNA in sera from patients with nasopharyngeal carcinoma. AB - BACKGROUND: The detection of tumor-derived DNA within the circulation of patients with malignant disease using polymerase chain reaction (PCR)-based strategies has opened a new avenue for the diagnosis and monitoring of these patients. Because of the universal association of Epstein-Barr virus (EBV) with the nonsquamous type of nasopharyngeal carcinoma (NPC; World Health Organization types II and III), the detection of cell free EBV DNA in sera from patients with NPC may be a valuable tool for monitoring the progress of tumors or to provide advanced warning of tumor recurrence. METHODS: Serum samples were obtained from different patients, and cell free EBV DNA was detected with a conventional PCR approach. A total of 134 patients were sampled, including 36 patients with primary NPC, 28 control patients, 18 patients suffering from locoregional recurrence, 7 patients with distant metastasis, and 45 patients with NPC in clinical remission. A conventional PCR approach employing standard 35-cycle and 50-cycle reactions was used to detect cell free EBV genomes. Results from the two PCR cycles were compared to provide a semiquantitative picture of the relative quantity of EBV genome in each serum sample. RESULTS: The EBV DNA detection rates, i.e., the rates of positive detection, for 35-cycle and 50-cycle PCR analyses, respectively, were 38.9% and 75% for patients with primary NPC, 3.5% and 10.7% for control patients, 27.8% and 88.9% for patients with locoregional disease recurrence, 71.4% and 100% for patients with distant metastasis, and 7.1% and 36.5% for patients with disease in clinical remission. The rates of positive detection among patients with active disease all appeared to be significantly greater compared with the rates among patients with disease in clinical remission. Longitudinal data for six patients with recurrent tumors revealed a close correlation between the relative quantity of circulating cell free EBV genomes and the disease course of these patients. The sensitivity, specificity, positive predictive value, and negative predictive value of the 50-cycle PCR analysis for detecting recurrent disease were 92%, 63.5%, 42.6%, and 96.4%, respectively. CONCLUSIONS: This study demonstrated that, by using a 50-cycle PCR based approach, high sensitivity and high negative predictive value for detecting recurrent disease can be obtained from the detection of the cell free EBV genome in sera from patients with NPC. The 50-cycle PCR analysis, therefore, may provide a simple, clinically useful adjuvant method for monitoring patients with NPC. PMID- 11857306 TI - National Cancer Data Base report on malignant paragangliomas of the head and neck. AB - BACKGROUND: Malignant paragangliomas of the head and neck are rare, with previous reports limited to nine or fewer patients. The current review included 59 cases extracted from the National Cancer Data Base that were diagnosed between 1985 1996. METHODS: The primary criterion for inclusion in the current study was verified metastatic spread from a paraganglioma of the head and neck. Patterns of presentation and treatment as well as clinically relevant associations were demonstrated in contingency tables. Relative survival was used for analysis of outcome. RESULTS: The average patient age at presentation was 44 years, and gender distribution was equivalent. Metastases were confined to regional lymph nodes in the majority of cases (68.6%), with carotid body tumors found to have an even higher rate of regional confinement (93.8%). Surgery was the most common treatment (76.3%). The use of adjuvant irradiation for regionally confined disease increased across time, from 27% (1985-1990) to 46% (1991-1996). The 5 year relative survival rate was 59.5% (76.8% for regionally confined carcinoma and 11.8% for distant metastasis). Among patients who were followed until death, those treated with adjuvant irradiation had a longer median survival (45 months) compared with those patients who were treated with surgery alone (12 months). CONCLUSIONS: Malignant paraganglioma represents metastatic spread of a tumor type that, when restricted to the site of origin, is considered benign. Metastases from malignant paragangliomas of the head and neck usually are regionally confined. The primary management of a recognized malignancy should be directed toward complete surgical removal of the primary tumor and regional lymph nodes. Postoperative irradiation may be beneficial in slowing the progression of residual disease. PMID- 11857307 TI - Angiogenesis in patients with craniopharyngiomas: correlation with treatment and outcome. AB - BACKGROUND: Craniopharyngiomas are histologically benign epithelial neoplasms of the sellar region that often exhibit aggressive and invasive growth. The authors hypothesized that tumor proliferation, spread, and recurrence are angiogenesis dependent and investigated the significance of vascularization relative to biologic behavior. To the authors' knowledge, angiogenesis for patients with craniopharyngiomas has not been examined to date. METHODS: The authors measured microvessel densities in resected, histologically proven craniopharyngiomas using immunostains for CD-34, a monoclonal antibody that selectively recognizes endothelial cells. Both histologic types of craniopharyngiomas, adamantinomatous and papillary, were included in the study. In addition, the cellular distribution of vascular endothelial growth factor (VEGF), a strong stimulator of new vessel formation, was assessed by both immunohistochemistry and in situ hybridization for VEGF receptor 2 (VEGFR-2) mRNA expression. RESULTS: Histologically, small numbers of capillaries were identified in temporal stroma but not in their epithelial components. Immunohistochemistry revealed strong, conclusive cytoplasmic immunoreactivity for VEGF in the epithelial cells of both adamantinomatous craniopharyngiomas and papillary craniopharyngiomas. In situ hybridization showed that VEGFR-2 mRNA was expressed widely, not only in neoplastic epithelium but also in capillary endothelium. CONCLUSIONS: Tumors with greater microvessel density regrow more frequently compared with tumors that have lower microvessel density, suggesting that the extent of angiogenesis is of prognostic value in patients with craniopharyngioma. VEGFR-2 may act as a key modulator of VEGF activity in endothelial cells and nonendothelial cells, indicating that VEGF plays an important role in the behavior of craniopharyngiomas. PMID- 11857308 TI - Chromosomal imbalances detected by comparative genomic hybridization are associated with outcome of patients with hepatocellular carcinoma. AB - BACKGROUND: Biologic characteristics of tumors are greatly affected by genetic aberrations. However, to the authors' knowledge there is no study that shows that cytogenetic information is useful for estimating prognosis of patients with hepatocellular carcinoma (HCC). METHODS: Comparative genomic hybridization (CGH) analysis was performed in 41 HCCs to examine whether the analysis of cytogenetic aberrations allows us to estimate biologic behavior of HCC. RESULTS: Tumor recurrence was linked to the loss at 13q (P = 0.0027) and to the number of DNA copy number aberrations (DCNAs; P = 0.0003). The decrease in DNA copy number at 8p and 13q and amplification at 11q13 were significantly associated with unfavorable outcome of patients (P = 0.017, P = 0.012, and P = 0.00081, respectively). The number of DCNAs was significantly different between favorable and poor prognosis patients with HCC; 5.78 +/- 2.7 versus 11.13 +/- 4.8 (P = 0.004), and it was an independent prognostic marker in HCCs. CONCLUSIONS: The current study indicates that cytogenetic information provided by CGH is useful for estimating prognosis of patients with HCC. PMID- 11857309 TI - Increased expression of beta-catenin predicts better prognosis in nonsmall cell lung carcinomas. AB - BACKGROUND: beta-Catenin has been shown to function as a Wnt signaling molecule to stimulate cyclin D1 expression and cell growth in several kinds of tumors. METHODS: The authors immunohistochemically examined specimens of 217 surgically resected primary nonsmall cell lung carcinomas (NSCLCs) for beta-catenin expression and classified them semiquantitatively into three categories, including those with high, moderate, and low scores of expression. RESULTS: High, moderate, and low scores of expression were found in 37 (17.1%), 145 (66.8%), and 35 (16.1%) tumors, respectively. beta-Catenin expression was not correlated with cyclin D1 expression, but was positively correlated with the Ki-67 cell growth fraction (P = 0.04). The direct sequencing analysis for the beta-catenin gene mutation of 13 specimens of 217 tumors for the current study revealed no mutations. The relation between survival and beta-catenin expression was evaluated in 148 potentially curatively resected tumors with pathologic Stages I IIIA. A trend toward better survival was found in patients with tumors having higher scores. In multivariate analysis, high beta-catenin expression was a significant and independent favorable prognostic factor (hazards ratio, 0.31; P = 0.007) as was pathologic stage. Analyzed by cell type, in nonsquamous cell carcinomas, patients with tumors having high scores survived a significantly longer time than those with tumors having moderate or low scores (5-year survival rates, 84%, 55%, and 32%, respectively; P = 0.02), and high beta-catenin expression tended to be a favorable prognostic factor (hazards ratio, 0.32; P = 0.052). CONCLUSIONS: These results indicate that, in NSCLCs, increased expression of beta-catenin can predict favorable prognosis of patients with resected tumors, suggesting that accumulation of beta-catenin has no or little oncogenic effect via activation of the Wnt pathway, unlike in colon carcinomas or hepatomas. PMID- 11857310 TI - Brain metastases from esophageal carcinoma: natural history, prognostic factors, and outcome. AB - BACKGROUND: Brain metastases from esophageal carcinoma are extremely rare, and information regarding the natural history, results of treatment, and possible prognostic factors in these patients is limited. METHODS: The records of 36 patients with brain metastases from esophageal carcinoma who were treated between 1986 and 2000 were reviewed. For brain metastases, 12 patients (33%) were treated with surgical resection followed by radiation therapy (S+RT), and the remaining 24 patients were treated with radiation therapy alone. RESULTS: At the initial diagnosis of esophageal carcinoma, the median primary tumor length was 8 cm (range, 2-19 cm), and 26 of 32 available patients (81%) had clinical Stage III-IV tumors according to the International Union Against Cancer 1997 criteria. At time brain metastases appeared, lung metastases were not demonstrated in 25 of 36 patients (69%) who were assessed by chest computed tomography (CT) scans. The overall median survival for all patients was 3.9 months (range, 0.6-36.8 months), and the actuarial survival rates at 12 months and 24 months were 14% and 3%, respectively. In univariate analysis, treatment modality, Karnofsky performance status (KPS), and extracranial disease status each had a statistically significant impact on survival, and, in multivariate analysis, treatment modality and KPS were statistically significant prognostic factors for survival. Five patients (14%) survived more than 1 year, all of whom were treated with S+RT. These five patients had inactive extracranial disease and, four of five patients (80%) had a 90-100% KPS. CONCLUSIONS: Brain metastases from esophageal carcinoma tended to occur in patients with a large primary tumors and/or disease in advanced clinical stages. With the appearance of brain metastases, an absence of lung metastasis frequently was observed on chest CT scans. The prognoses for these patients were generally poor, although selected patients may survive longer with intensive brain tumor treatment. PMID- 11857311 TI - Stromelysin-3 is expressed by aggressive meningiomas. AB - BACKGROUND: Stromelysin-3 (ST3) that belongs to the metalloproteinase family is proposed to play a role in tumor invasion. The purpose of this work was to study the variation of ST3 expression in meningiomas. METHODS: Twenty atypical meningiomas were retrieved from the Pathology Department's files at Hopital de Bellevue, Saint-Etienne, France. They were compared with 20 benign meningiomas randomly selected from the same file. The tumors were classified using standard histologic criteria. Frozen sections of the tumors were immunostained for ST3 and MIB-1 to evaluate the proliferative activity of tumor cells. RESULTS: The study included 5 fibrous meningiomas, 10 transitional meningiomas, 20 syncitial meningiomas, 2 secretory meningiomas, 2 microcystic meningiomas, and 1 angiomatous meningioma. Stromelysin-3 was expressed within the stromal and neoplastic cells of only 1 benign meningioma and 13 atypical meningiomas. The MIB 1 proliferation index was significantly higher in the meningiomas expressing ST3 (Student t test: P < 0.001). The invasion of bone, muscle, and brain by meningiomas as well the recurrence were statistically correlated with their ST3 expression (Kruskal-Wallis nonparametric correlation test, P = 0.001 and P = 0.008, respectively). CONCLUSIONS: Stromelysin-3 might play an important role in the invasiveness of meningiomas. Therefore, considering, ST3 in association with evaluation of the MIB-1 proliferating index may be an useful tool to assess the behavior of meningiomas. PMID- 11857312 TI - Central nervous system relapse in adults with acute lymphoblastic leukemia. AB - BACKGROUND: Recurrence of acute lymphoblastic leukemia (ALL) in the central nervous system (CNS) in adults is considered a poor prognostic feature. Few reports have analyzed this issue. The objective of this study was to analyze the experience with CNS recurrence in adult patients with ALL. METHODS: Between December 1982 and April 2000, the records of 527 consecutive, newly diagnosed patients with ALL who were treated at the M. D. Anderson Cancer Center were reviewed. Patients were treated with one of two published regimens: vincristine, doxorubicin, and dexamethasone (VAD; n = 237 patients) or hyperfractionated cyclophosphamide plus VAD (hyper-CVAD) (n = 290 patients). Among 439 patients (83%) who achieved complete remission (CR), 32 patients (7%) had a CNS recurrence. All patients had received CNS prophylaxis tailored to the risk of CNS recurrence with systemic chemotherapy with or without intrathecal (IT) chemotherapy; none received cranial radiotherapy for prophylaxis. The records of these patients were reviewed. RESULTS: The median age at the time of diagnosis was 33 years. The median CR duration prior to CNS recurrence was 36 weeks (range, 2-185 weeks). Three groups were identified: 1) patients with an isolated CNS recurrence (n = 17 patients), 2) patients with CNS recurrences after bone marrow (BM) recurrence (n = 8 patients), and 3) patients with simultaneous CNS and BM recurrences (n = 7 patients). IT chemotherapy was effective in achieving a CNS CR in 30 patients (94%), but 10 patients (31%) had a second CNS recurrence. The median survival from CNS recurrence was 6 months; 28% of patients were alive at 1 year, and 6% of patients were alive at 4 years. CONCLUSIONS: Adults with ALL and CNS recurrences have a poor prognosis despite effective IT chemotherapy. Future studies should investigate better approaches in the treatment of these patients to improve their long-term survival. Effective CNS prophylaxis remains the single best approach for treating patients with CNS leukemia. PMID- 11857313 TI - Impaired platelet aggregation in melanoma patients treated with interferon-alpha 2b adjuvant therapy. AB - BACKGROUND: High-dose interferon (INF)- alpha-2b is the only Food and Drug Administration-approved adjuvant treatment for patients with melanoma who are at high risk of recurrence. Although circumstantial evidence points to a potentially harmful effect of INF-alpha-2b on platelet function, to the authors' knowledge this has never been studied in humans. METHODS: The study group was comprised of patients who had undergone surgery for melanoma and were free of disease but at a high risk of recurrence. All patients were candidates for adjuvant INF treatment (high-dose) and were undergoing routine evaluation to which platelet aggregation was added. Aggregation was triggered in standard fashion with adenosine diphosphate, epinephrine, collagen, thrombin, arachidonic acid, and ristocetin. Blood samples were drawn immediately before treatment, during the intravenous loading phase, during the subcutaneous maintenance phase, and 3-6 weeks after cessation of treatment. Patients receiving low-dose, long-standing INF-alpha-2b treatment also were tested. All results at each phase were compared with those of normal controls. RESULTS: In those patients receiving high-dose INF-alpha-2b, ristocetin-induced aggregation did not appear to be affected. However, the response to > or = 1 of the other agonists was impaired in 5 of 6 samples during loading, 14 of 15 samples during the maintenance phase, and 8 of 13 samples after treatment, compared with only 1 of 8 samples before treatment (P = 0.025, P = 0.002, and P = 0.067, respectively). During treatment with low-dose INF, platelet function was affected to a lesser extent. CONCLUSIONS: INF treatment in melanoma patients appears to be associated with severe impairment of platelet aggregation, which appears to be dose-dependent and cumulative-dose-dependent. This is not detectable by the standard coagulation profile. This effect has significant implications in the event of accidental injury or elective surgery. The antiaggregation activity may be the mechanism by which INF delays, reduces, or prevents the formation of melanoma metastases. PMID- 11857314 TI - Chemotherapy, irradiation, and surgery for function-preserving therapy of primary extremity soft tissue sarcomas: initial treatment with ifosfamide, mitomycin, doxorubicin, and cisplatin plus granulocyte macrophage-colony-stimulating factor. AB - BACKGROUND: Most institutional teams utilize multimodality therapy in their efforts to cure patients with primary high-grade extremity soft tissue sarcomas, although the value of adjuvant systemic chemotherapy is still disputed by some oncologists. This single-institution Phase II study describes an effort to control metastasis by the use of two cycles of chemotherapy as the initial preoperative treatment. METHODS: Between March 1994 and October 1997, 20 women and 19 men with primary extremity or limb girdle high-grade soft tissue sarcomas were registered to a study of preoperative ifosfamide, mitomycin, doxorubicin, cisplatin (IMAP) plus granulocyte macrophage-colony-stimulating factor (GM-CSF) followed by preoperative irradiation and subsequent limb-sparing surgery. The two sequential monthly cycles of IMAP involved intravenous ifosfamide, 2500 mg/m(2), and mesna, 2500 mg/m(2), on Day 0, followed by identical doses of these agents plus intravenous mitomycin, 4 mg/m(2), doxorubicin, 40 mg/m(2), and cisplatin, 60 mg/m(2), on Day 1. Sargramostim (GM-CSF) 250 microg/m(2) was given subcutaneously every 12 hours for 4 days beginning 6 days before the chemotherapy, and then for 14 more days beginning the day after chemotherapy was completed. At the beginning of the third month, external beam irradiation was administered daily, 5 days each week for 5 consecutive weeks to total preoperative doses of 4500 centigrays (cGy). This was accompanied by reduced doses of MAP chemotherapy (mitomycin, 6 mg/m(2), doxorubicin, 30 mg/m(2), and cisplatin, 45 mg/m(2)) intravenously on Days 0, 21, and 42 of the radiation therapy segment. Approximately 1 month after preoperative irradiation ended, each patient had complete surgical excision with curative intent, using limb-sparing techniques when possible. Radiation to total doses of 5500-6500 cGy was accomplished by delivery of an additional 1000-2000 cGy to the tumor bed via intraoperative electron beam, brachytherapy, or external beam irradiation at the completion of surgery. RESULTS: All except 5 patients had tumors at least 5 cm in diameter. Chemotherapy toxicity grade three or higher consisted primarily of vomiting (23%), leukopenia (54%), and thrombocytopenia (77%). Six patients have died of metastatic sarcoma, and one other died in a motorcycle accident. Kaplan-Meier curves indicate estimated 5-year survival of approximately 80% and freedom from metastasis at 2 years of approximately 85%. CONCLUSIONS: IMAP plus GM-CSF is satisfactory as initial treatment for primary extremity soft tissue sarcomas in two monthly cycles preceding irradiation. The prescribed irradiation was generally tolerable and effective in permitting limb sparing surgery. Although the outcome of patients treated on this regimen has been favorable, the metastasis problem has not been eliminated. PMID- 11857315 TI - Loss of heterozygosity on chromosome arms 3p and 6q in microdissected adenocarcinomas of the uterine cervix and adenocarcinoma in situ. AB - BACKGROUND: Despite the increasing frequency of adenocarcinomas of the uterine cervix, little is known regarding inactivation of tumor suppressor genes (TSGs) in this tumor type. The authors analyzed loss of heterozygosity (LOH) in 36 carcinomas of the cervix with glandular differentiation, and 5 adenocarcinoma in situ in 40 patients. METHODS: The authors analyzed samples using laser capture microdissection from archival material and DNA amplified with microsatellite markers on the following loci: 3p14.2 (D3S1234, D3S1300), 3p21.3 (D3S1029, D3S1447), 3p22-24 (D3S1537, D3S1351), 6q21-23.3 (D6S250), 6q25.1 (ESR), 6q25.2 (D6S255), 8p21 (D8S136, D8S1820), 13q12.3 (D13S220, D13S267), 17q21 (D17S579, D17S855). Eight additional markers spanning the short arm of chromosome 3 (3p12 p25) and six spanning the long arm of chromosome 6 (6q11-q27) were studied in the cases showing LOH to further define the deletion intervals. RESULTS: The frequency of allelic loss in cancers was chromosome 3p: 49% (p14.2: 35%, p21.3: 23%, p22-24: 41%), 6q: 48% (q21-23.1: 39%, q25.1: 45%, q25.2: 7%), 13q: 22%, 17q: 6%, and 8p: 18%. On chromosome arm 3p, the authors' data suggest at least two discrete areas of deletion: a proximal area between markers D3S1234 (p12) and D3S1766 (p14.2-14.3), and a second distal interval, telomeric from marker D3S4623 (p21.3). On chromosome 6q, the deletion area is between marker D6S300 (q22) and D6S255 (q25.2). Two of five preneoplastic lesions showed LOH on chromosome arm 3p, and two five showed allelic loss on chromosome arm on 6q, suggesting the genes might be inactivated early in cervical tumorigenesis. CONCLUSIONS: The authors have identified three chromosomal regions that may harbor TSGs involved in the development/progression of adenocarcinomas of the uterine cervix, 3p12 14.2, 3p21.3-pter, and 6q22-25.2. Deletions also were detected in adenocarcinoma in situ, suggesting the genes may be inactivated early in cervical tumorigenesis. PMID- 11857316 TI - Immunologic quantitation of the carcinoma specific human carcinoma antigen in clinical samples. AB - BACKGROUND: Based on the cross-reactivity of the human carcinoma antigen, HCA, with epiglycanin, a mouse mammary carcinoma cell surface glycoprotein, HCA has been detected in the tissue and blood of patients with every type of epithelium derived cancer tested. METHODS: Competitive binding assays utilized the following antiepiglycanin antibodies: a polyclonal rabbit antiserum (immunoglobulin [Ig] G and IgM) in a radioimmunoassay; mouse monoclonal antibodies (Ab-1, IgM) on immunoplates; anti-idiotypic (Ab-2) and anti-anti-idiotypic (Ab-3) monoclonal antibodies (both IgG) from spleen cells of C57BL mice immunized, respectively, with Ab-1 and Ab-2, and utilized on immunoplates. IgG and IgM antibodies were evaluated for their ability to detect HCA and to distinguish between the blood of patients with, or without, carcinomas. RESULTS: Assays with the rabbit antiserum distinguished plasmas of metastatic breast carcinoma patients from those of patients with benign breast disease with a sensitivity of approximately 93% (specificity 90%). Antiepiglycanin IgM monoclonal antibodies (i.e., AE3) showed high specificity and sensitivity (> 90%) with sera from advanced carcinoma patients when compared with normal sera. The IgG anti-anti-idiotypic (Ab-3) monoclonal antibodies (i.e., AF2), which bind the same epitope as Ab-1, appear to possess less nonspecific binding capacity, however, than the Ab-1 (IgM) antibodies. Anti-Ab-1 (i.e., C8F2) anti-idiotypic monoclonal antibodies, which bear an idiotope equivalent to the epitope present in epiglycanin and the HCA, demonstrated greater consistency as a standard calibrator and for coating wells than epiglycanin. CONCLUSIONS: The concentration of the HCA in the body fluids of patients with carcinomas may be accurately determined by competitive binding assays. It is suggested that the use of anti-idiotypic antibodies (IgG), rather than epiglycanin/HCA, and Ab-3 anti-anti-idiotypic antibodies (IgG), rather than Ab-1 (IgM), will improve the consistency, as well as the sensitivity and specificity, of the assay. PMID- 11857317 TI - Expression of CD95 (Fas) in sun-exposed human skin and cutaneous carcinomas. AB - BACKGROUND: Carcinomas of the skin are by far the most common human malignancies. Continuous exposure to ultraviolet (UV) light facilitates the development of precancerous lesions (actinic keratosis [AK]) that may progress to invasive squamous carcinomas. Apoptosis, triggered by the activation of CD95 (Fas), is one of the most important defense mechanisms against UV light-induced carcinogenesis in experimental models, but the dynamics of CD95 expression in patients with sun induced lesions are largely unknown. METHODS: The authors studied the expression of CD95 (Fas) in biopsy samples of normal skin (not exposed to sun) and compared it with chronically sun-exposed skin (as evidenced by solar elastosis), AK, squamous cell carcinoma (SCC) and basal cell carcinoma (BCC), and keratoacanthomas (KA). RESULTS: Normal skin keratinocytes expressed CD95 in cytoplasmic membranes and intercellular bridges in the basal layer. In chronically sun-exposed keratinocytes (solar elastosis, no evidence of dysplasia), CD95 expression was up-regulated and was observed throughout the entire thickness of the epidermis. However, in actinic keratosis there was a complete absence of Fas in approximately two-thirds of the cases (8 of 12). In invasive SCC, CD95 was expressed focally and weakly only at the sites of contact with stromal lymphocytes. Keratoacanthomas consistently expressed CD95 at the interface with the inflammatory cells. No staining was observed in BCC. CONCLUSIONS: CD95 (Fas) up-regulation in chronically sun-exposed keratinocytes indicates an important role in the control of sun-induced damage. Further sun exposure results, however, in significant down-regulation of this defense mechanism, proportional to the degree of dysplasia. PMID- 11857318 TI - Cytokeratin expression patterns in noncardia, intestinal metaplasia-associated gastric adenocarcinoma: implication for the evaluation of intestinal metaplasia and tumors at the esophagogastric junction. AB - BACKGROUND: Barrett esophagus (BE)/Barrett adenocarcinoma and distal gastric intestinal metaplasia (IM)/adenocarcinoma are similar histologically, but they differ in their clinical presentation, epidemiology, and pathogenesis. Differentiating BE from gastric IM and Barrett adenocarcinoma from gastric adenocarcinoma is difficult, especially when IM is short or tumors are large and involve both sides of the esophagogastric junction. Previously, the authors identified unique cytokeratin (CK) immunoreactivity patterns that were associated strongly with BE and Barrett adenocarcinoma. The specificity of CK7 and CK20 (CK7/20) expression patterns in patients with IM-associated gastric adenocarcinoma, which is distinct epidemiologically from BE/Barrett adenocarcinoma, has not been evaluated. The objective of the current study was to evaluate the CK7/20 expression patterns in noncardia, IM-associated gastric adenocarcinoma in a Chinese population with a low risk for BE and esophageal adenocarcinoma and a high risk for Helicobacter pylori infection and gastric carcinoma. METHODS: Endoscopic biopsy specimens of gastric IM and adjacent tumor from 50 consecutive patients with advanced noncardia gastric carcinoma were immunostained for CK7 and CK20. Clinical and endoscopic features and H. pylori status were documented. Two gastrointestinal pathologists, blinded to clinical and endoscopic data, independently assessed CK7/20 immunohistochemistry. RESULTS: H. pylori infection was present in 43 of 50 patients (86%). In the area of IM, patchy CK7 staining was seen in 9 patients (18%), and diffuse CK20 staining was seen in all 50 patients (100%). The BE CK7/20 pattern characterized by CK7 staining in superficial and deep glands and the CK20 staining in surface epithelium was not seen in any of the 50 patients. Only one patient (2%) demonstrated a CK7 positive/CK20 negative immunophenotype characteristic of Barrett adenocarcinoma. The remaining 49 patients (98%) showed non-Barrett adenocarcinoma patterns of CK7/20 staining, i.e., a CK7 positive/CK20 positive pattern was seen in 33 patients (66%), a CK7 negative/CK20 positive pattern was seen in 12 patients (24%), and a CK7 negative/CK20 negative pattern was seen in 4 patients (8%). CONCLUSIONS: In a patient population without risk factors for the development of BE/esophageal adenocarcinoma, the CK7/20 pattern characteristic of BE was not present in gastric IM adjacent to adenocarcinoma, and the CK7/20 pattern characteristic of Barrett adenocarcinoma also was extremely rare. These results support the hypothesis that, despite the presence of intestinalized mucosa in both disorders, BE/Barrett adenocarcinoma and gastric IM/adenocarcinoma are two distinct clinical entities with unique demographic, clinical, and CK immunophenotypic findings. These results may have application to the evaluation of patients with IM and adenocarcinoma arising at the esophagogastric junction. PMID- 11857320 TI - Alternative bimonthly cycles of doxorubicin, cyclophosphamide, and etoposide, cisplatin with hematopoietic growth factor support in patients with carcinoma of unknown primary site. AB - BACKGROUND: Because carcinomas of unknown primary origin are highly malignant tumors with a bad prognosis (median survival, 6-12 months) and no current optimal therapy, the authors designed a prospective dose-dense chemotherapy regimen with the objective of improving the results observed in patients who receive conventional treatment. METHODS: Eighty-two patients received alternative bimonthly cycles of doxorubicin 50 mg/m(2) with cyclophosphamide 1000 mg/m(2) (AC) and etoposide 300 mg/m(2) with cisplatin 100 mg/m(2) (EP). Cycles were given at 2-week intervals with granulocyte-macrophage-stimulating factor support (5 microg/kg per day) from Day 4 to Day 10. Patients without measurable lesions were included, because the major end point was survival. RESULTS: The median number of alternative cycles of AC and EP was 4 cycles (range, 1-12 cycles). An objective response was observed in 24 of 62 patients (39%; 95% confidence interval, 30-48%) with measurable lesions, including 6 patients who achieved a complete response. Among 20 patients with nonmeasurable disease, 9 patients (45%) had no evidence of progressive disease at the end of chemotherapy. The overall median survival of 82 patients was 10 months, with 5 patients surviving clinically disease free at 17 months, 29 months, 45 months, 64 months, and 70 months after the end of treatment. Myelosuppression was the most common toxicity. Two toxic deaths occurred. CONCLUSIONS: Using these doses and schedules, a dose-dense chemotherapy regimen did not appear to improve the outcome of patients with carcinoma of unknown primary site. Alternative studies dealing with news drugs will be required. PMID- 11857319 TI - Episodic (breakthrough) pain: consensus conference of an expert working group of the European Association for Palliative Care. AB - BACKGROUND: Breakthrough pain is transitory exacerbation of pain that occurs in addition to otherwise stable persistent pain. The wide differences in estimation of incidence reported in literature are probably because of different settings and meanings attributed to the definition of breakthrough pain. METHODS: A panel of experts met to establish the actual knowledge on breakthrough pain, according to the evidence in literature and experience. They agreed that episodic or transient pain could be a more simple and adequate term in most languages, including English, French, Italian, and Spanish. RESULTS: A specific assessment and precise pain characterization are essential to plan the most appropriate treatments. Despite the relevance of this temporal pain pattern for the influence on the outcome and quality of life, few controlled studies have been performed to give evidence of a specific approach. Several experiences have reported the possible efficacy of different drugs, route of administration, and modalities of administration in different circumstances. CONCLUSIONS: Prospective studies with previous treatments using similar terminologies are necessary to find the most convenient therapeutic intervention, according to the temporal pattern characteristics and the pain mechanism involved. PMID- 11857321 TI - Severe interstitial pneumonitis associated with docetaxel administration. AB - BACKGROUND: Interstitial pneumonitis has not been reported as a toxicity of docetaxel. The authors report the presentation and natural history of four patients who developed a severe interstitial pneumonitis after receiving docetaxel. METHODS: The hospital and outpatient records of patients treated with docetaxel were reviewed to identify whether any of these patients required an evaluation for respiratory problems. RESULTS: Four patients developed an interstitial pneumonitis that could be explained only as a toxicity of docetaxel. None had metastatic disease to the lung, and all had normal liver function before receiving chemotherapy. The patients presented with acute dyspnea and fever within 1-2 weeks of receiving docetaxel. All developed progressive interstitial infiltrates and respiratory failure that required mechanical ventilation. An exhaustive workup for other causes of pneumonitis was negative. Broad-spectrum antibiotics and corticosteroids were ineffective. Two patients died of complications related to the pulmonary process. The two survivors required ventilatory support for more than 21 days. The clinical and pathologic findings of these patients are presented. CONCLUSIONS: Interstitial pneumonitis is a rare and potentially fatal complication of docetaxel treatment. Prolonged ventilatory support is appropriate in patients with a favorable prognosis. PMID- 11857322 TI - Proliferation marker KI-S5 discriminates between favorable and adverse prognosis in advanced stages of neuroblastoma with and without MYCN amplification. AB - BACKGROUND: The biologic behavior of neuroblastoma is notoriously variable, and even carefully elaborated prognostic models fail to predict the clinical course in a portion of cases. Because the proliferative activity is determined by the sum of all molecular imbalances that influence cell cycling, the authors investigated the potential prognostic relevance of the tumor growth fraction in neuroblastoma. METHODS: A retrospective analysis was conducted on a cohort of 161 neuroblastoma patients with a median follow-up period of 72.8 months. Tumors were classified according to Hughes typing and grading criteria. The proliferative index (PI) was assessed immunohistochemically on archival biopsy specimens using monoclonal antibody Ki-S5 (Ki-67), and the MYCN status was determined by means of Southern blot analysis. RESULTS: The PI, MYCN status, International Neuroblastoma Staging System (INSS) stage, International Neuroblastoma Pathology Classification grade, Hughes grade, and the patients' age at diagnosis were all found to be significant predictors of event free survival by univariate Kaplan-Meier analysis. However, the PI identified prognostically distinct subsets in higher tumor stages and Grade 2 and 3 neuroblastomas as well as tumors with unfavorable histology, and enabled risk stratification in tumors with and without MYCN amplification (P = 0.034 and 0.002, respectively). Multivariate Cox regression analysis selected INSS stage (relative risk [RR], 4.05; P < 0.0001) and the PI (RR, 2.49; P = 0.007) as the sole independent prognostic indicators, whereas MYCN entered the selection only after exclusion of the PI. CONCLUSIONS: It emerges that the PI as a single factor has greater predictive power than the MYCN status. Proliferation measurements therefore might significantly improve the accuracy of current prognostic models for neuroblastoma. PMID- 11857323 TI - The association of treatment-related symptoms with quality-of-life outcomes for localized prostate carcinoma patients. AB - BACKGROUND: Most studies of treatment outcomes in men with localized prostate carcinoma have emphasized sexual, urinary, and bowel symptoms with the assumption that they have an impact on quality of life. However, very few studies have directly examined and compared the impact of these symptoms on overall and cancer specific quality of life. METHODS: The authors examined 783 incident cases of localized prostate carcinoma, diagnosed from 1993 to 1998, and 1928 age-matched healthy controls from the Health Professionals Follow-Up Study cohort. Information on frequency of ejaculation and urinary symptoms were collected before cancer diagnosis. After cancer diagnosis, the authors mailed a questionnaire including the Medical Outcomes Study Short Form-36 Health Status Survey (SF-36), the Cancer Rehabilitation Evaluation System-Short Form (CARES SF), and the University of California at Los Angeles Prostate Cancer Index in 1998. RESULTS: Cases had slightly lower scores on most of the SF-36 scales and reported much more bother from sexual, urinary, and bowel symptoms compared with healthy controls. Among prostate carcinoma patients, bowel symptoms had the greatest negative impact on quality of life, followed by sexual and urinary symptoms. As expected, treatment-related symptoms were associated with the physical domains of quality of life, but psychosocial domains were just as strongly affected. CONCLUSIONS: Patients and health care providers need to consider the potential mental quality-of-life impacts associated with prostate carcinoma treatment symptoms when making treatment decisions. Even after patients have completed cancer treatment, significant health impairments may remain. Health care providers should continue to address the mental and physical well being of prostate carcinoma patients in follow-up care. PMID- 11857324 TI - Development of rats' maternally directed orienting behaviors from birth to day 2. AB - We previously observed 2- to 3-day-old (P2-3) rats perform maternally directed orienting behaviors (MDOBs)-locomotion, torso flexion, rotation to the supine orientation, and snout probing, accompanied by audible vocalizations (barking) under surfaces having properties of the dam's ventrum, which result in close contact between the pup's ventrum and the surface. We now report on the development of MDOBs from birth through P2. Wistar rats (24 litters) were tested at one of four times: 1 hr after birth before the first nursing bout (P0pre), on the day of birth after the first nursing bout (P0post), on postnatal Day 1 (P1), or on postnatal Day 2 (P2). Pups were placed prone on a warmed platform inside a corral where, for 4 min, they encountered one of two motherlike surfaces at head height: (a) a soft cotton roof or (b) a patch of the pup's own anesthetized dam's ventrum. Control pups received no overhead stimulus. The behaviors we had identified as MDOBs and others, including limb behaviors, are easily recognized at P0pre, and, in aggregate, are more frequent then than at any later age. The differential responsiveness of most of the behaviors to the dam's ventrum compared to the cotton roof or open condition is relatively low at P0pre, but rises rapidly through P2. Behavioral structure develops quickly from a rudimentary principal component with two behaviors at P0pre to two principal components incorporating seven behavior classes by P2. MDOBs are discussed in relation to late-term fetal behaviors, early postnatal righting responses, and the oral nipple grasp, and as a model of the development of filial attachment in an altricial mammal. PMID- 11857325 TI - Classical conditioning in the rat fetus: involvement of mu and kappa opioid systems in the conditioned response. AB - When the Embryonic Day 20 (E20) rat fetus is given a conditioning trial involving a paired presentation of an artificial nipple (the conditioned stimulus; CS) with an intraoral infusion of milk (the unconditioned stimulus; US), it shows evidence of classical conditioning when again exposed to the CS during a test trial. Specifically, the fetus shows fewer oral grasp responses (the conditioned response; CR) when continuously presented with the artificial nipple. The present study further investigated this classically conditioned reduction in oral grasping. Separate experiments (a) examined the time course of the reduction in oral grasping (Experiment 1), (b) characterized the time course of mu opioid (Experiment 2) and kappa opioid (Experiment 3) involvement in the CR, and (c) described changes in fetal behavior (Experiment 4) associated with mu and kappa opioid effects on responding to the artificial nipple. The results are discussed in terms of opioid involvement in establishing and maintaining early suckling behavior. PMID- 11857327 TI - The interaction of age and unconditioned stimulus intensity on long-trace conditioned flavor aversion in rats. AB - To see if the neural representation of the conditioned stimulus (CS) is available to old-age rats beyond the time it is available to young adults, the intensity of the unconditioned stimulus (US) and the length of the CS-US interval were systematically varied in a trace conditioning experiment. Results indicated that increasing US intensity extends the interval over which trace conditioning is evident in old-age rats but not in young adults, suggesting that trace decay occurs more rapidly in young rats. Results were interpreted in terms of age differences in the workings of hypothesized biochemical timing mechanisms that may directly influence the ability to associate stimuli over trace intervals in conditioned taste-aversion procedures. PMID- 11857326 TI - Classical conditioning in the rat fetus: temporal characteristics and behavioral correlates of the conditioned response. AB - This study examined the temporal characteristics and behavioral correlates of the conditioned response (CR) following classical conditioning of the embryonic Day 20 (E20 rat fetus). The conditioning procedure involved pairing of an artificial nipple (the CS) with an infusion of milk (the US) to establish classical conditioning. The test for classical conditioning involved measurement of a stimulus-evoked facial wiping response in a classical conditioning test. Experiment 1 compared the effectiveness of one- and three-trial procedures to establish classical conditioning. Experiment 2, 3, and 4 described the time course for the CR following one- and three-trial conditioning procedures. Experiments 3b and 4b describe the behavioral responses to (a) presentation of the CS at the time of conditioning, (b) infusion of the milk US at the time of conditioning, and (c) reexposure to the CS before the test for classical conditioning. Experiments 5 and 6 exposed the fetus to manipulations that either increased or decreased stretching (a behavior found to be associated with the CR). The results are discussed in terms of the temporal characteristics and behavioral correlates of conditioned and unconditioned responses and their mediation by activity in endogenous mu and kappa opioid systems. PMID- 11857328 TI - Functional recovery and dendritic hypertrophy after posterior and complete cingulate lesions on postnatal day 10. AB - Rats received lesions of the posterior cingulate cortex or both the anterior and posterior cingulate cortex (total cingulate), or sham procedures, on postnatal Day 10. As adults, animals were trained in the Morris water task. Both the cingulate lesion groups showed substantial functional recovery relative to our previous studies of adult operates or animals with perinatal cingulate lesions. A Golgi analyis of layer III pyramidal cells in parietal cortex showed an increase in dendritic length in the lesion animals relative to sham controls, which is similar to previous findings for rats with anterior cingulate but not motor or parietal lesions. In addition, there was a partial regeneration of the anterior tissue in the total cingulates, which in some cases extended into the posterior region. This is consistent with earlier findings that anterior cingulate lesions around Day 10 stimulate neurogenesis. It appears that there is something special about the reparative processes and subsequent functional recovery that follow midline neocortical lesions. PMID- 11857329 TI - Developmental differences in temporal patterns and potentiation of isolation induced ultrasonic vocalizations: influence of temperature variables. AB - The present study examined the relationship between the thermal environment and core body temperature in producing age-related patterns of ultrasonic vocalizations (USVs). Implanted telemetry devices allowed on-line measurement of core body temperature during an extended period of isolation and after maternal contact, both as a function of age and thermal environment. At 12 or 17 days of age, rat pups were isolated for 30 min in either a cool or a warm environment, returned home for 5 min, and then re-isolated for 10 min. Number of USVs, body temperature, and behavioral activity were measured. During initial isolation in a cool environment, 12-day-olds displayed relatively stable patterns of ultrasounding and body temperature across time whereas older animals showed a time-dependent increase in USV calling and in core temperature. During re isolation, 12-day-olds potentiated their USV calling at both ambient temperatures while 17-day-olds did not. The overall results suggest a strong dependence between USV calling, core body temperature, and ambient temperature during initial isolation at both ages-a finding in agreement with interpretations of USVs as an acoustic by-product of thermal challenge. In contrast, during re isolation, USV calls, core body temperature, and ambient temperature functioned independently-a finding in agreement with interpretations of USVs as a representation of an emotional state. PMID- 11857330 TI - Gastric saline infusion reduces ultrasonic vocalizations and brown fat activity in suckling rat pups. AB - Under standard conditions involving isolation and cooling, it has been documented that intraoral infusion of milk and injection of the intestinal peptide cholecystokinin (CCK) result in an attenuation in ultrasonic vocalizations (USV) emitted by infant rat pups. One of the most effective stimuli in inhibiting ingestion in suckling rat pups is gastric distension, but the effect of gastric distension on USV production has not been reported. In this experiment, we subjected infant rats to intragastric infusion of isotonic saline (2% body weight) to produce a natural level of gastric distension and hydration. We found that this stimulus resulted in a powerful reduction in USV emissions in isolated 10-day-old rats. In a subsequent experiment, we found that gastric saline infusion also diminished brown adipose tissue (BAT) thermogenesis. There were different time courses of the gastric saline infusion effects on BAT thermogenesis and on USV emissions, however, suggesting that these processes may be independently regulated. We hypothesize that this stimulus induces a transient activation of the parasympathetic nervous system, which overrides the sympathetic control of BAT and USV production. PMID- 11857331 TI - Facilitating postural control: effects on the reaching behavior of 6-month-old infants. AB - In this study, 3-D kinematic as well as 2-D videorecordings were made of the reaching behavior of infants aged about 6 months who were not yet able to sit. Detailed analyses of these recordings were directed toward specifying the effects of providing additional postural support to the lower body on the spatial and temporal features of such behavior. To detect these effects, reaching and associated head movements in this modified condition were compared to those made while the infants sat in an age-appropriate and commercially available chair lacking the supplementation of support for the pelvic region and upper legs. Findings consistent with predictions included better head stabilization and smoother reaching movements when the infants were in the modified chair. In addition, these two achievements were negatively related to reaching experience. These, and other findings, underscore the infrequently investigated supposition that changes in postural control induce improvements in the control of reaching movements during infancy. Recommendations are made about how the procedure adopted in the present study could be used in subsequent research to give further insights into the codevelopment of posture and action. PMID- 11857332 TI - Measuring infant memory: does the ruler matter? AB - Although the mobile conjugate reinforcement, deferred imitation, and visual attention paradigms are commonly used to test memory processing by infants, there is often debate about the comparability of memory measured by these three procedures. In a series of experiments, we systematically compared 6-month-old infants' performance across all three procedures. Contrary to current claims, measures of memory obtained using the mobile conjugate reinforcement and deferred imitation paradigms were more similar than those obtained using the deferred imitation and visual attention paradigms. PMID- 11857333 TI - Ligands for murine NKG2D display heterogeneous binding behavior. AB - NKG2D transmits stimulatory signals to natural killer cells and other hematopoietic cells, leading to enhanced proliferation, cytokine secretion and target killing. Murine and human NKG2D each recognize five known class I-related molecules with distinct primary structures. Here, we used surface plasmon resonance to examine the binding of murine NKG2D to its cognate ligands: RAE-1B6 (a newly described C57BL/6J variant of RAE-1), RAE-1 delta (common to BALB and C57BL6/J), and H60 (expressed in BALB, but not C57BL/6J). While RAE-1B6 and H60 display relatively high affinities for NKG2D with K(D) in the 20-30 nM range and k(off )in the 0.03s(-1) to 0.06s(-1) range (t(1/2) approximately 10-20s); the RAE 1 delta variant binds with a lower affinity: K(D) of approximately 750 nM. Furthermore, RAE-1 delta displays biphasic kinetics with dominant k(off) of approximately 0.2s(-1) (t(1/2) approximately 3s), partially explaining the lower affinity. Thus, H60 and RAE-1B6 bind NKG2D with almost identical kinetics while sharing only 20% amino acid sequence identity; whereas other RAE-1 molecules demonstrate faster dissociation and lower affinities than RAE-1B6 despite sharing 90% sequence identity. C57BL/6J mice, although not expressing the H60 gene product, retain a high-affinity ligand for NKG2D in the form of RAE-1B6. PMID- 11857335 TI - Endo- and exocytic rate constants for spontaneous and protein kinase C-activated T cell receptor cycling. AB - To determine the rate constants of spontaneous and activated TCR cycling, we examined TCR endo- and exocytosis in the human T cell line Jurkat by three different methods. Using a simple kinetic model for TCR cycling and non-linear regression analyses, we found that the spontaneous endocytic rate constant of the TCR was low (approximately 0.012 min(-1)) whereas the spontaneous exocytic rate constant was similar to that of other cycling receptors (approximately 0.055 min( 1)). Following protein kinase C activation (PKC) the endocytic rate constant was increased tenfold (to approximately 0.128 min(-1)) whereas the exocytic rate constant was unaffected. Thus, the TCR becomes a rapidly cycling receptor with kinetics similar to classical cycling receptors subsequent to PKC activation. This results in a reduction of the half-life of cell surface expressed TCR from approximately 58 to 6 min and allows rapid redistribution of the TCR during T cell activation. PMID- 11857334 TI - Cellular sources and targets of IFN-gamma-mediated protection against viral demyelination and neurological deficits. AB - IFN-gamma is an anti-viral and immunomodulatory cytokine critical for resistance to multiple pathogens. Using mice with targeted disruption of the gene for IFN gamma, we previously demonstrated that this cytokine is critical for resistance to viral persistence and demyelination in the Theiler's virus model of multiple sclerosis. During viral infections, IFN-gamma is produced by natural killer (NK) cells, CD4(+) and CD8(+) T cells; however, the proportions of lymphocyte subsets responding to virus infection influences the contributions to IFN-gamma-mediated protection. To determine the lymphocyte subsets that produce IFN-gamma to maintain resistance, we used adoptive transfer strategies to generate mice with lymphocyte-specific deficiencies in IFN-gamma-production. We demonstrate that IFN gamma production by both CD4(+) and CD8(+) T cell subsets is critical for resistance to Theiler's murine encephalomyelitis virus (TMEV)-induced demyelination and neurological disease, and that CD4(+) T cells make a greater contribution to IFN-gamma-mediated protection. To determine the cellular targets of IFN-gamma-mediated responses, we used adoptive transfer studies and bone marrow chimerism to generate mice in which either hematopoietic or somatic cells lacked the ability to express IFN-gamma receptor. We demonstrate that IFN-gamma receptor must be present on central nervous system glia, but not bone marrow derived lymphocytes, in order to maintain resistance to TMEV-induced demyelination. PMID- 11857336 TI - Pathogenic autoantibody production requires loss of tolerance against desmoglein 3 in both T and B cells in experimental pemphigus vulgaris. AB - Mechanisms of tolerance break against desmoglein 3 (Dsg3) in patients with pemphigus vulgaris (PV) producing pathogenic anti-Dsg3 IgG autoantibodies are unclear. In this study, using a novel PV mouse model involving Dsg3 knockout mice, we investigated the mechanisms leading to production of autoantibodies against Dsg3. Adoptive transfer of Dsg3(-/-) splenocytes immunized with recombinant mouse Dsg3 to Rag2(-/-) recipient mice expressing Dsg3 resulted in the stable production of anti-Dsg3 IgG and development of PV phenotypes including oral erosions with suprabasilar acantholysis. When purified T and B cells from Dsg3(-/-), Dsg3(+/-) or Dsg3(+/+) mice were mixed with various combinations and transferred to Rag2(-/-) mice, pathogenic anti-Dsg3 IgG production was observed only with a combination of Dsg3(-/-) T and Dsg3(-/-) B cells but not with the other combinations. These results suggest that loss of tolerance against Dsg3 in both B and T cells is important for the development of autoimmune state of PV. PMID- 11857337 TI - PD-1:PD-L inhibitory pathway affects both CD4(+) and CD8(+) T cells and is overcome by IL-2. AB - Programmed death-1 (PD-1) is an immunoreceptor tyrosine-based inhibitory motif (ITIM)-containing receptor expressed upon T cell activation. PD-1(-/-) animals develop autoimmune diseases, suggesting an inhibitory role for PD-1 in immune responses. Members of the B7 family, PD-L1 and PD-L2, are ligands for PD-1. This study examines the functional consequences of PD-1:PD-L engagement on murine CD4 and CD8 T cells and shows that these interactions result in inhibition of proliferation and cytokine production. T cells stimulated with anti-CD3/PD-L1.Fc coated beads display dramatically decreased proliferation and IL-2 production, while CSFE analysis shows fewer cells cycling and a slower division rate. Costimulation with soluble anti-CD28 mAb can overcome PD-1-mediated inhibition by augmenting IL-2 production. However, PD-1:PD-L interactions inhibit IL-2 production even in the presence of costimulation and, thus, after prolonged activation, the PD-1:PD-L inhibitory pathway dominates. Exogenous IL-2 is able to overcome PD-L1-mediated inhibition at all times, indicating that cells maintain IL-2 responsiveness. Experiments using TCR transgenic CD4(+) or CD8(+) T cells stimulated with antigen-presenting cells expressing PD-L1 show that both T cell subsets are susceptible to this inhibitory pathway. However, CD8(+) T cells may be more sensitive to modulation by the PD-1:PD-L pathway because of their intrinsic inability to produce significant levels of IL-2. PMID- 11857338 TI - Transmission of antibody-induced arthritis is independent of complement component 4 (C4) and the complement receptors 1 and 2 (CD21/35). AB - The K/BxN murine model of rheumatoid arthritis (RA) is dependent on the specificity of the KRN alpha beta-TCR, to recognize glucose-6-phosphate-isomerase (GPI) on the NOD MHC class II A(g7) allele and production of GPI-specific autoantibodies. Transfer of K/BxN serum into MHC-unrelated and lymphocyte deficient mice induces RA. To investigate whether K/BxN serum-induced RA involves complement activation and/or the complement receptors (CR) 1 and 2, we analyzed the role of complement C4 and of CR1 and CR2. For this purpose we used C4(-/-) mice impaired in the classical and the lectin complement pathways; Cr2(-/-) mice lacking CR1 and CR2 and, as control strains, BALB/c, C57BL/6, KRN and NOD. RA was assessed by caliper measurement of ankle thickness, clinical index and joint histology. We found that all mouse strains except NOD developed RA. The lack of protection in C4(-/-) mice suggests that antibody-mediated RA is independent of the classical as well as the lectin complement pathways and the split complement product C4b. The lack of protection in Cr2(-/-) mice suggests that absence of CR1 had no significant affect, considering its role in immune complex clearance, inhibition of C3 and C5 convertase and as receptor for C3b/C4b. Also, CR2 lacks a role in disease as analyzed here, in its possible functions as receptor for C3dg, germinal center reaction and activation of alternative pathway on binding iC3. Hence we conclude that the transmission of K/BxN serum-induced RA is independent of the classical and the lectin complement pathways and CR1 and CR2. The crucial role of complement C5, while neither classical nor lectin pathway is necessary, indicates that the alternative complement pathway may have a role in the K/BxN serum-induced RA model. PMID- 11857341 TI - The neutrophil-activating protein (HP-NAP) of Helicobacter pylori is a potent stimulant of mast cells. AB - Infection by Helicobacter pylori causes an acute inflammatory response followed by a chronic infection of the human gastric mucosa. A neutrophil-activating protein (HP-NAP) has been identified in H.pylori, and its role in infection and immune response is currently under investigation. Here, we show that HP-NAP induces beta-hexosaminidase release and interleukin-6 production in peritoneal mast cells, two actions which are completely inhibited by pertussis toxin. We also show that in polarized epithelial cell monolayers HP-NAP translocates from the apical to the basolateral domain, where mast cells are located. These findings characterize HP-NAP as an inflammatory factor of H.pylori that is effective from the beginning of the inflammatory cascade. PMID- 11857339 TI - Cellular and molecular requirements for the recall of IL-4-producing memory CD4(+)CD45RO(+)CD27(-) T cells during protection induced by attenuated Plasmodium falciparum sporozoites. AB - The requirements for maintenance of antigen (Ag)-specific memory T cells in protection to malaria is poorly understood. We have previously demonstrated a recall of IL-4-producing memory CD4(+)CD45RO(+) T cells with parasitized red blood cells (pRBC) in persons protected by radiation-attenuated Plasmodium falciparum sporozoites (gamma-spz). Using the CD27 marker, we have now identified two subsets of CD4(+)CD45RO(+) T cells: CD4(+)CD45RO(+)CD27(+) T cells representing an early memory and CD4(+)CD45RO(+)CD27() T cells representing a terminally differentiated memory cells. A small subset of CD4(+)CD45RO(+)CD27(-) T cells also expressed CD70, the CD27 ligand. The addition of anti-CD70 monoclonal antibody (mAb) to pRBC-stimulated cultures significantly inhibited the conversion of CD27(+) to CD27(-) subset without profoundly affecting IL-4 production. In contrast, the inclusion of anti-CD27 mAb in parallel cultures abrogated IL-4 production without interfering with conscription of T cells into the CD27(-) T cell set. We propose that the persistence of memory CD4(+) T cells depends on Ag-driven conscription of a mature memory phenotype through co ligation of CD27 and CD70 expressed, respectively, on CD27(+) and CD27(-) T cells. Hence, protracted protection in malaria depends in part on memory CD4(+) T cells that require specific Ag presumably from the repositories of liver-and blood-stage antigens and the delivery of a second signal from the CD27:CD70 interaction. PMID- 11857342 TI - Hypo-active variant of IL-2 and associated decreased T cell activation contribute to impaired apoptosis in autoimmune prone MRL mice. AB - Apoptosis of activated lymphocytes is crucial to the maintenance of immune homeostasis and self-tolerance, as demonstrated by the well-known autoimmune MRL lpr mouse lacking the death receptor Fas. However, even MRL+/+ activated T cells have a resistance to Fas-mediated apoptosis as compared to T cells from the non autoimmune FVB/N strain. To understand the molecular mechanisms underlying these strain differences, we studied biochemical characteristics of T cells upon activation. Compared to FVB/N T cells, MRL T cells under-expressed procaspase-3 but over-expressed FLIP(L). In addition, up-regulation of Bcl-x(L), IL-2, and CD25 was diminished in MRL cells, suggesting inadequate T cell activation. Upon finding that MRL, like other autoimmune strains NOD and SJL, has a hypo-active variant of the IL-2 gene, we added wild-type murine recombinant (mr)IL-2 during activation. Exogenous mrIL-2 restored MRL apoptosis to the level of FVB/N; in addition, expression of procaspase-3, and FLIP(L), Bcl-x(L) and CD25 was normalized. These results suggest that defective MRL T cell activation, in part due to hypo-active IL-2, underlies the impaired apoptosis of this strain. In addition, the hypo-active variant of IL-2 shared among autoimmune strains may, by causing diminished cell activation and cell death, predispose these strains to develop autoimmune disease. PMID- 11857340 TI - Relationship between kinetic stability and immunogenicity of HLA-DR4/peptide complexes. AB - Immunodominant T cell epitopes from the autoantigen human cartilage glycoprotein 39 have previously been mapped in the context of HLA-DR*0401 and *0402, using mice expressing HLA-DR4 transgenes. We measured the dissociation rates of these epitopes from soluble recombinant DR*0401 and DR*0402 to assess the relationship between peptide/HLA-DR4 kinetic stability and immunogenicity. Experiments were performed at endosomal pH (5.5) and at cell surface pH (7), in the absence and presence of soluble recombinant HLA-DM (sDM). All (4/4) immunodominant peptide/HLA-DR complexes exhibit dissociation half-times of 1h to several days. In contrast, most (3/4) non-immunodominant complexes dissociate with half-times <30 min under at least one of these conditions. Interestingly, a complex which is stable except in the presence of HLA-DM at pH 5.5 is immunogenic only following peptide immunization, while a complex which is stable at acidic but not at neutral pH, is non-immunogenic following either whole protein or peptide immunization. These data indicate that kinetic stability of peptide/MHC complexes in vivo is a key determinant of immunogenicity. PMID- 11857343 TI - A subfraction of B220(+) cells in murine bone marrow and spleen does not belong to the B cell lineage but has dendritic cell characteristics. AB - Although CD45R/B220 is commonly used as a pan-B cell marker in the mouse, not all B220(+) cells belong to the B cell lineage. Here we report the characterization of a subpopulation of B220(+)CD19(-) cells in murine bone marrow, which failed to express markers that are present in early CD19(--) B cell precursors. Instead, these cells expressed low levels of MHC class II and CD11c, which are typically found on dendritic cells (DC). Moreover, these B220(+)CD19(-)CD11c(+) cells expressed Gr-1, indicating that they are related to the recently identified murine plasmacytoid DC or their progenitors. Therefore, we evaluated surface marker expression of the B220(+)CD19(-)CD11c(+) cells in lymphoid tissues of C57BL/6 mice, recombinase activating gene-1 deficient mice, lacking mature B and T lymphocytes, and mice with a targeted disruption of the Ig H chain mu membrane exon (mu MT), lacking mature B lymphocytes. When comparing bone marrow and spleen, we found that the surface profiles of B220(+)CD19(-)CD11c(+) cells were remarkably similar, indicating that they are in a comparable maturation or activation stage in the two lymphoid compartments. In addition, the almost complete absence of peripheral B220(+) B-lineage cells in mu MT mice allowed the anatomical localization of the B220(+)CD19(-)CD11c(+) cells to the red pulp and the T cell areas in the spleen. Taken together, our findings indicate that the mouse bone marrow contains a recirculating population of B220(+)CD19(-) CD11c(+) plasmacytoid DC, the development of which is largely independent of the presence of mature T and B cells. PMID- 11857344 TI - Requirement for both IL-12 and IFN-gamma signaling pathways in optimal IFN-gamma production by human T cells. AB - Phytohemagglutinin (PHA)-derived T lymphoblasts or T cell clones from patients genetically deficient in IL-12R beta 1 (IL-12R beta 1(-/-)) or IFN-gamma R1 (IFN gamma R1(-/-)) produced two- to threefold reduced IFN-gamma levels compared to the corresponding cells from healthy individuals after anti-CD3 and PMA stimulation. Moderate IFN-gamma production was observed in PHA-derived T lymphoblasts or T cell clones derived from healthy subjects in the presence of anti-IFN-gamma R1 or anti-IL-12 mAb, whereas it was negligible in the presence of both mAb. However, when anti-IFN-gamma R1 and/or anti-IL-12 mAb were added during restimulation, the cells produced normal levels of IFN-gamma, indicating that both IFN-gamma and IL-12 had an effect on the priming phase. Moderate production of IFN-gamma was partially enhanced only in IFN-gamma R1(-/-) T cell clones generated in the presence of IL-12, but was almost completely abolished when IL 12R beta 1(-/-) and IFN-gamma R1(-/-) T cell clones were generated in the presence of anti-IFN-gamma R1 or anti-IL-12 mAb, respectively. IL-4 production was enhanced in T cell clones from IL-12R beta 1(-/-),but not from IFN-gamma R1( /-) patients, whereas IL-10 and IL-2 production did not differ significantly in polyclonal T cells or clones from healthy and deficient individuals. These results indicate that IL-12R beta 1- and IFN-gamma R1-dependent signals co ordinately regulate IFN-gamma, but not IL-2 and IL-10 production, whereas only IL 12 negatively controls IL-4 production by in vitro-generated T cell clones. Thus, although IL-12 and IFN-gamma signals are each sufficient for moderate production of IFN-gamma by human T cells, both are needed for optimal IFN-gamma production, and in the absence of both IFN-gamma production is completely abrogated. PMID- 11857345 TI - Transformation of the microvascular system during multistage tumorigenesis. AB - Simian virus SV40 large T Antigen expression in the islets of Langerhans of transgenic mice results in beta-cell hyperproliferation, onset of new blood vessel formation and the development of highly vascularized solid tumors. Angiogenesis in the RIPTag mouse model, as well as in human cancer, is a hallmark of multistage tumorigenesis and precedes the development of solid tumors. In our study, intravital microscopy was used to monitor changes in the blood vessel phenotype, microcirculation and leukocyte adhesion during the progression from normal islets to angiogenic islets and solid tumors. In RIP1-Tag5 mice, an aberrant microangioarchitecture becomes apparent in early stages during spontaneous tumor development. Notably, the transition from normal to angiogenic islets is characterized by an increase in vessel diameter rather than vessel numbers. Thus, dilatation of existing vessels precedes vessel sprouting. Once initiated, neovascularization in angiogenic islets results in loss of vessel hierarchy and differentiation. Solid insulinomas display a higher vessel density and even more dramatic vessel heterogeneity as revealed by local "hot spots" of neovascularization and irregular vessel diameters. Strikingly, profound changes in the microangioarchitecture are already observed in early angiogenic islets suggesting that key features of the angiogenic vasculature are established prior to the expansion of tumor mass. Moreover, adhesion of leukocytes was found to be dramatically decreased in both angiogenic islets and solid tumors and correlates with morphological alterations of the vasculature. Thus, vessel transformation and reduced leukocyte-endothelium interactions are not exclusively features of solid tumors but represent early events during tumorigenesis. PMID- 11857346 TI - TCL-1, MTCP-1 and TML-1 gene expression profile in non-leukemic clonal proliferations associated with ataxia-telangiectasia. AB - We analyzed the role of 4 genes, TCL-1, MTCP-1, TML-1 and ATM, in the early pathogenesis of T cell leukemia, with particular interest in the characteristics of long-standing non-leukemic clonal proliferations in ataxia-telangiectasia (A T) patients. Five patients were studied: 4 patients had A-T (2 of whom had non leukemic clonal proliferations [ATCP]), 1 had B cell lymphoma and 1 had T-ALL; a fifth patient with T-PLL did not have A-T. We measured the levels of expression for TCL-1, MTCP-1 and TML-1. TCL-1, not expressed in unstimulated mature T cells, was upregulated in the peripheral blood leukocytes (PBL) of the 2 A-T patients with ATCP. It was also expressed in the malignant cells of the A-T patient with B cell lymphoma and the T-PLL cells of the patient without A-T. In the same cells, MTCP-1 type A was expressed equally in all 5 patients, as well as in the controls; MTCP-1 type B transcripts were not observed. TML-1, also not expressed in unstimulated T cells, was expressed in the PBL of one A-T patient with ATCP and in the leukemic cells of the non-A-T T-PLL patient. These expression patterns were compared to cellular immunophenotypes. The non-leukemic clonal T cell populations had the characteristics of immature T cells. We conclude that TCL-1 and TML-1 play a role in cell proliferation and survival but are not pivotal genes in the progression to malignancy, even when the ATM gene is mutated. Additional genetic alterations must occur to initiate tumorigenesis. PMID- 11857347 TI - Human papillomavirus type 16 E6-enhanced susceptibility to apoptosis induced by TNF in A2780 human ovarian cancer cell line. AB - In our study, we show that expression of HPV-16 E6 sensitizes TNF-induced cytotoxicity of human ovarian cancer cell line A2780. This effect is not related to a different number of TNF receptors present on cell membrane. The major induction of massive apoptosis induced by TNF is not p53- and p21(waf-1) dependent but it is principally related to NF-kappaB inhibition in A2780/E6 cells. Consistently to NF-kappaB inhibition a rapidly release of cytochrome c and severe induction of DNA fragmentation are seen in A2780/E6 cells. Also in human colon cancer cell line HCT-116/E6 the expression of HPV-16 E6 enhances TNF cytotoxicity. This effect is not present in the HCT-116/mu-p53 clone (transfected with a dominant-negative mutated p53 transgene). Thus, taken together all these observations suggest that HPV-16 E6 sensitizes A2780 and HCT-116 cells to TNF; this effect is not p53-dependent, but it is essentially mediated through an inhibition in activating NF-kappaB activities. PMID- 11857348 TI - Extra-cellular thiol metabolism in clones of human metastatic melanoma with different gamma-glutamyl transpeptidase expression: implications for cell response to platinum-based drugs. AB - Thiol redox status can affect important functions both intracellularly and extracellularly. The plasma membrane enzyme gamma-glutamyl transpeptidase (GGT), which plays a crucial role in cellular handling of thiols, is often expressed in malignant tumors, including melanoma, although its expression levels may vary widely among different tumors or cells of the same tumor. In an attempt to better understand the functional significance of GGT overexpression, we have examined the relationships between intra- and extra-cellular thiol metabolism and GGT expression. Intra- and extra-cellular distribution of glutathione and other low mol. wt. thiols and disulfides was investigated in two different Me665/2 human melanoma clones that originated from the same metastasis, but exhibiting high (2/60 clone) and low (2/21 clone) GGT activity. Intracellular content of glutathione was lower in GGT-rich 2/60 cells, in spite of high GGT expression. A lower utilization of extracellular cystine was also observed in these cells. In both clones, a direct secretion of cysteine in the extracellular medium was detected, which was independent of GGT-mediated catabolism of extracellular glutathione. Substantial amounts of glutathione, GSSG and glutathione-cysteine disulfide were accumulated extracellularly only in the case of GGT-poor 2/21 cells, while the same event was apparent in 2/60 cells only after the following inhibition of GGT activity. When exposed to the trinuclear platinum compound BBR 3464 or hydrogen peroxide, which are very reactive for sulfur-containing nucleophiles, the 2/60 clone showed higher sensitivity than the 2/21 clone to both agents. These results suggest that the clone-specific balance between transport of sulfur aminoacids and GGT activity results in profound differences in the capability of each clone to modify the thiol redox status of the extracellular milieu. The finding may have important implications in tumor cell behavior with particular reference to chemosensitivity, since thiols are recognized factors in modulation of cell sensitivity to platinum-based anticancer drugs. PMID- 11857349 TI - Potentiating antitumor effects of a combination therapy with lovastatin and butyrate in the Lewis lung carcinoma model in mice. AB - Lovastatin, the drug used for the treatment of hypercholesterolemia, has previously been reported to exert antitumor activity in experimental murine models. Butyrate and butyric acid derivatives are well known to induce differentiation and apoptosis of tumour cells and also have recently gained acceptance as potential anticancer agents. In this study, we examined the antitumor effects of the combination of lovastatin and butyrate or its prodrug tributyrin in vitro and in vivo against a murine Lewis lung carcinoma (3LL). This combination therapy showed synergistic antitumor activity against 3LL cells in vitro. These effects were at least in part due to apoptosis induction that occurred after 12 hr of incubation with lovastatin and butyrate and was preceded by changes in cell cycle distribution of treated cells and expression of p21, p53 and cyclin D1. Remarkably, a systemic treatment of syngeneic mice inoculated with 3LL cells with both drugs resulted in significant tumour growth retardation. PMID- 11857351 TI - Cellular effects of purvalanol A: a specific inhibitor of cyclin-dependent kinase activities. AB - We have studied the effects of purvalanol A on the cell cycle progression, proliferation and viability. In synchronized cells, purvalanol A induced a reversible arrest the progression in G1 and G2 phase of the cell cycle, but did not prevent the completion of DNA synthesis in S-phase cells. The specificity of action of the drug was supported by the selective inhibition of the phosphorylation of cyclin-dependent kinase (cdk) substrates such as Rb and cyclin E. The cell contents of cyclins D1 and E were lower in cells incubated with purvalanol A compared to controls, but the level of the cdk inhibitory protein p21(WAF1/CIP1) was increased, indicating that the drug did not cause a general inhibition of gene expression. Purvalanol A did not inhibit transcription under cell-free conditions. This compound, however, caused an inhibition of the estradiol-induced expression of an integrated luciferase gene, suggesting that cdk or related enzymes may participate in the regulation of the activity of certain promoters. When exponentially growing cells, both mouse fibroblasts and human cancer cell lines, were incubated with purvalanol A for prolonged periods of time (24 hr), a lasting inhibition of cell proliferation as well as cell death were observed. In contrast, a 24 hr incubation of quiescent (non-transformed) cells with purvalanol A did not prevent their resumption of cell cycle after removal of the drug. PMID- 11857350 TI - Impact of BCRP/MXR, MRP1 and MDR1/P-Glycoprotein on thermoresistant variants of atypical and classical multidrug resistant cancer cells. AB - The impact of the ABC transporters breast cancer resistance protein/mitoxantrone resistance associated transporter (BCRP/MXR), multidrug resistance-associated protein 1 (MRP1) and multidrug resistance gene-1/P-glycoprotein (MDR1/PGP) on the multidrug resistance (MDR) phenotype in chemoresistance and thermoresistance was investigated in the parental human gastric carcinoma cell line EPG85-257P, the atypical MDR subline EPG85-257RNOV, the classical MDR subline EPG85-257RDB and their thermoresistant counterparts EPG85-257P-TR, EPG85-257RNOV-TR and EPG85 257RDB-TR. Within the atypical MDR subline EPG85-257RNOV expression of BCRP/MXR and of MRP1 were clearly enhanced (vs. parental and classical MDR lines). MDR1/PGP expression was distinctly elevated in the classical MDR subline EPG85 257RDB (vs. parental and atypical MDR sublines). In all thermoresistant counterparts basal expression of BCRP/MXR, MRP1 and MDR1/PGP was increased relative to thermosensitive sublines. Although it could be shown that the overexpressed ABC transporters were functionally active, however, no decreased drug accumulations of doxorubicin, mitoxantrone and rhodamine 123 were observed. Thus, expression of BCRP/MXR, MRP1 and MDR1/PGP was found to be dependent on the appropriate type of chemoresistance; correlating with a classical or atypical MDR phenotype. Within the thermoresistant variants, however, the increase in ABC transporter expression did obviously not influence the MDR phenotype. PMID- 11857352 TI - Role of stromal collagen in immunomodulation and prognosis of advanced gastric carcinoma. AB - Although several hypotheses have been proposed explaining the mechanisms of the immune-privileged status of malignant tumors, the exact pathway is yet to be explored. Tumor stroma plays a vital role in the prognosis of cancer patients; however, the immunomodulatory impact of gastric cancer stroma has not been reported. We have evaluated the amount of stromal collagen and its impact on the infiltration of immune-competent cells into the tumor cell nest in gastric carcinoma. Tissue specimens from 84 advanced gastric carcinoma patients who had undergone a curative resection were evaluated for host immune status (CD8+ T cells), tumor stromal reaction (AZAN staining), tumor Fas ligand expression and incidence of tumor cell apoptosis (by TUNEL). The number of apoptotic tumor cells (apoptotic index [AI]) increased proportionally with an increase in the number of CD8+ T cells within the cancer cell nest (nest CD8) (p = 0.0001). Nest CD8 was inversely correlated with the amount of stromal collagen (p < 0.0001). Nest CD8 and AI became independent predictors of patient survival (p = 0.0023 and p = 0.044, respectively) in Cox's multivariate analysis. The amount of stromal collagen was found to be a significant predictor of disease relapse in univariate analysis (p = 0.0010) but not in multivariate analysis (p = 0.4729). In conclusion, increased nest CD8 produced a survival advantage by inducing tumor cell apoptosis in gastric carcinoma patients. Increased tumor stromal collagen worked as a barrier for CD8+ T-cell infiltration and might be one of the mechanisms of tumor escape from the host immune attack. PMID- 11857353 TI - Lysosomal proteases as potential targets for the induction of apoptotic cell death in human neuroblastomas. AB - Neuroblastoma is the most common type of cancer in infants. In children this tumor is particularly aggressive; despite various new therapeutic approaches, it is associated with poor prognosis. Given the importance of endosomal-lysosomal proteolysis in cellular metabolism, we hypothesized that inhibition of lysosomal protease would impact negatively on neuroblastoma cell survival. Treatment with E 64 or CA074Me (2 specific inhibitors of cathepsin B) or with pepstatin A (a specific inhibitor of cathepsin D) was cytotoxic for 2 neuroblastoma cell lines having different degrees of malignancy. Cell death was associated with condensation and fragmentation of chromatin and externalization of plasma membrane phosphatidylserine, 2 hallmarks of apoptosis. Concomitant inhibition of the caspase cascade protected neuroblastoma cells from cathepsin inhibitor induced cytotoxicity. These data indicate that prolonged inhibition of the lysosomal proteolytic pathway is incompatible with cell survival, leading to apoptosis of neuroblastoma cells, and that the cathepsin-mediated and caspase mediated proteolytic systems are connected and cooperate in the regulation of such an event. Since modern antitumor chemotherapy is aimed at restoring the normal rate of apoptosis in neoplastic tissues, the demonstration that endosomal lysosomal cathepsins are involved in this process may constitute a basis for novel strategies that include cathepsin inhibitors in the therapeutic regimen. PMID- 11857355 TI - No association between HER-2 gene polymorphism at codon 655 and a risk of bladder cancer. AB - The amplification and overexpression of the human epidermal growth factor receptor 2 gene HER-2 (also known as c-erb-B2 or neu) have been shown to be associated with bladder cancer and its progression. Recent studies indicated an association between the Ile to Val polymorphism at codon 655 of HER-2 and susceptibility to breast cancer. To investigate the correlation between the Ile/Val polymorphism and the susceptibility and progression of bladder cancer, we analyzed the polymorphism in 232 patients with transitional cell carcinoma of the bladder and 408 normal controls. The frequencies of the Ile/Ile, Ile/Val and Val/Val genotype were 75.9%, 21.6% and 2.6%, respectively, in patients with bladder cancer and 75.7%, 23.0% and 1.2%, respectively, in controls. Statistical analyses of the genotype prevalence showed no significant difference between bladder cancer patients and normal controls (p = 0.419). Moreover, no significant differences in the genotype prevalence were observed when the patients were stratified according to the tumor grade, stage and smoking habits. When the Ile/Ile genotype was compared to the Ile/Val and Val/Val genotypes, a significant difference was found only between the patients with tumor stage Ta and those with T1-4 (age, gender and smoking habits-adjusted odds ratio = 2.13, 95% confidence interval = 1.09-4.15, p = 0.027). When the Ile/Ile + Ile/Val genotypes compared to the Val/Val genotype, no significant findings were observed. These results suggested that the HER-2 polymorphism at codon 655 is unlikely to be associated with the onset of bladder cancer. Furthermore, the findings suggest no association between this polymorphism and the disease progression in bladder cancer, although the possibility remains that the Ile/Ile genotype may be related to an increased risk of disease progression. PMID- 11857354 TI - Candidate tumor suppressor, HCCS-1, is downregulated in human cancers and induces apoptosis in cervical cancer. AB - To identify the genes involved in cervical carcinogenesis, we applied the mRNA differential display method and identified a candidate tumor suppressor gene, HCCS-1, which was present in normal cervical tissue but absent in cervical cancer, metastatic lymph node and CUMC-6 cervical cancer cell line. HCCS-1 transcripts were expressed in many normal tissues including leukocyte, lung, spleen, liver, heart and uterine cervix. Its expression was absent in 8 human cancer cell lines. HCCS-1-transfected HeLa cells exhibited growth inhibition by about 50%. This inhibitory effect of HCCS-1 on cervical cancer cells was associated with apoptotic process including DNA fragmentation. HCCS-1-transfected HeLa cells were shown to release cytochrome c from mitochondria, which activates caspase-9 and -3 and finally results in cleavage of poly(ADP-ribose) polymerase. Apoptosis formation was detected by propidium-iodide/annexin V. HCCS-1 transfected HeLa cells were more sensitive to adriamycin or UVC ray triggered apoptosis. These results suggest that HCCS-1 is downregulated in multiple human tumor types and may serve as a candidate tumor suppressor gene through apoptotic pathway against human cervical cancer. PMID- 11857356 TI - HIV-1 infection of primary effusion lymphoma cell line triggers Kaposi's sarcoma associated herpesvirus (KSHV) reactivation. AB - Kaposi's sarcoma-associated herpesvirus (KSHV) is a gammaherpesvirus etiologically linked to primary effusion lymphoma (PEL), to a subset of multicentric Castleman's disease and to Kaposi's sarcoma (KS), the most common neoplasm associated with AIDS. Among KSHV-infected individuals, the risk of KS is much higher in those with human immunodeficiency-1 (HIV-1) infection than among those with other types of immunosuppression, suggesting a direct action of HIV-1 on KSHV replication. We show in our report that BC-3 cells, a chronically KSHV infected B-cell line of a PEL origin, are permissive to HIV-1, offering a new tool for studying the interactions between these 2 viruses. In these cells, HIV-1 infection leads to reactivation of latent KSHV genomes, as demonstrated by the expression of KSHV lytic viral mRNAs. Although recombinant HIV-1 gp120 fails to enhance herpesvirus expression, transient transfection of the HIV-1 trans activator Tat suffices to reactivate latent KSHV. By showing that HIV-1 infection directly reactivates latent KSHV, our results suggest a direct role of HIV-1 in the onset of KS in coinfected individuals. PMID- 11857358 TI - Impact of the American anti-smoking campaign on lung cancer mortality. AB - Customary statistics on smoking practices are limited because they do not correlate well with the frequency of smoking-related diseases. Our study developed outcome measures based on lung cancer mortality and used them to assess the anti-smoking campaign. Changes in mortality from lung cancer were used to assess significant smoking among 5-year birth cohorts of white men born from 1901 to 1942. We used each cohort's lung cancer mortality rate at ages 40-44 to indicate its earlier smoking. A lung cancer mortality ratio was developed to describe each cohort's continued smoking from ages 40-44 to 55-59. These ratios were then compared with the durations of the cohorts' exposure to the anti smoking campaign that began in 1965. Lung cancer mortality in white men ages 40 44 peaked in 1970 and declined continuously thereafter, indicating that the anti smoking campaign promptly reduced significant smoking among younger men. However, the lung cancer mortality ratio indicates that only half of smokers in the specified birth cohorts were able to quit by ages 55-59, despite receiving ever more intense anti-smoking messages. The anti-smoking campaign produced moderate benefits among younger white male smokers but fewer benefits among older smokers because of the existence of a large number of inveterate smokers. PMID- 11857357 TI - Serologic response to human oncogenic papillomavirus types 16, 18, 31, 33, 39, 58 and 59 virus-like particles in colombian women with invasive cervical cancer. AB - The serologic response against virus-like particles (VLP) from 7 high risk genital papillomaviruses was investigated by ELISA in 147 Colombian women with invasive cervical cancer and 147 age-matched cytologically normal and HPV-DNA negative women. Anti-VLP antibodies were detected in 82% of the invasive cervical cancer patients and in 56% of the controls. Detection of antibodies against multiple HPV types is the rule and the presence of high antibody titers was associated with higher survival of cancer patients. Higher anti-VLP seroprevalence was observed in younger cancer patients. In those followed serologically for 1 year, antibodies generally remained at the same level. However, in some patients an increase or decrease in antibody levels occurred simultaneously for multiple HPV types, suggesting cross-reactivity between the HPV types investigated. Investigation of seroreactivity between 8 high risk HPVs suggested that there is some cross-reactivity between phylogeneticaly-related types such as 16, 31, 33 and 58; and 18, 45 and 59. In conclusion, our results confirmed (i) the high rate of HPV infections in Colombia, both in patients with cervical cancer and in the general population, and the particularly high rate of infections due to HPV 31 and 58; and (ii) the validity of anti-VLPs as a marker of present or past HPV infection. The simultaneous appearance or disappearance of antibodies against multiple HPV VLPs suggests that the antibodies detected by ELISA are not always type specific. PMID- 11857359 TI - Risk of pancreatic cancer among individuals with a family history of cancer of the pancreas. AB - In a hospital based case-control study of pancreatic cancer in Ontario and Quebec, a total of 174 incident pancreatic cancer cases and 136 healthy controls were compared for their family history of cancer. Information regarding the ages and sites of cancer was taken for 966 first-degree relatives of the cancer cases and for 903 first-degree relatives of the controls. A total of 150 cancer cases were reported among the relatives of the cases, compared to 122 cases among the relatives of the controls (relative risk 1.15; p = 0.23). Pancreatic cancer was the only site statistically in excess in the case relatives, compared to the control relatives (relative risk = 5.0; p = 0.01). The lifetime risk of pancreatic cancer was 4.7% for the first-degree relatives of the pancreatic cancer cases. The risk was 7.2% for relatives of cases diagnosed before age 60, and was 12.3% for relatives of patients with multiple primary cancers (all ages). These individuals comprise a high-risk group for pancreatic cancer and might benefit from enhanced surveillance or chemoprevention. Familial site-specific pancreatic cancer appears to be a distinct genetic entity, but contributes only modestly to the total burden of pancreatic cancer. PMID- 11857360 TI - Incidence and mortality from stomach cancer in Japan, Slovenia and the USA. AB - The mortality and incidence from stomach cancer were compared in Japan (a country with a high incidence where there was full application of mass screening during this period) and 2 countries with no screening policy: the USA (with a very low incidence) and Slovenia (with an intermediate rate). The registered cases of stomach cancer were from the Osaka Cancer Registry, the Slovenian National Cancer Registry and the Surveillance, Epidemiology, and End Results (SEER) registries in the USA. In the period 1975-95, the age-adjusted incidence rate (/100,000) of stomach cancer declined in the 3 countries, as follows: Japan, from 76.0 to 53.0 in men and 38.4 to 21.3 in women; Slovenia, from 40.2 to 24.1 in men and 16.6 to 10.8 in women; and the USA, from 9.5 to 6.9 in men and 4.3 to 2.9 in women. During the same period, the age-adjusted mortality rate declined, as follows: Japan, from 60.2 to 34.2 in men and 30.5 to 14.1 in women; Slovenia, from 37.7 to 21.2 in men and 13.8 to 9.0 in women; the USA, from 5.6 to 4.7 in men and 2.5 to 2.3 in women. In the period studied, specific trends on incidence and mortality with a cohort effect occurred only in Japan: analysis by the age period-cohort model confirmed that the decline has occurred since the generations born in 1910. The trend therefore corresponds to unplanned prevention through changes in environmental factors occurring since the early 20th century. The study of stage specific incidence rates confirmed the declining trend for regional cancer, whereas there was an increase in the incidence of localized cancer, associated with a period effect in 1975-95. This is attributed to the policy of early detection of stomach cancer, with the inclusion of intramucosal lesions of favorable prognosis and explains why mortality decreased faster than incidence during the period. PMID- 11857361 TI - Cancer incidence in families with multiple glioma patients. AB - Twenty-four Finnish families with 2 or more glioma patients were identified through questionnaires sent to 369 consecutive glioma patients receiving surgery at Tampere University Hospital during 1983-94. To explore whether unusual cancer susceptibility is involved, the cancer risk of 2,664 family members was estimated using population-based data from the Finnish Cancer Registry. Among the total cohort of relatives, 88 cancers were observed during 1953-97. The overall cancer risk among 12 families with juvenile onset gliomas was significantly decreased (standardized incidence ratio [SIR] 0.6, 95% confidence interval [CI]: 0.4-0.9). Among 12 families with adult onset gliomas, the overall cancer risk was equal to that of the reference population (SIR 1.1, 95% CI: 0.8-1.4) whereas the risk of skin melanoma (SIR 4.0, 95% CI: 1.5-8.8) and meningioma (SIR 5.5, 95% CI: 1.1-16) were significantly increased. Several other tumors, including those associated with neurofibromatosis 1 and 2, tuberous sclerosis and Li-Fraumeni and Turcot syndromes were surveyed, but no elevated risks were observed. In conclusion, the presence of meningiomas and skin melanomas in glioma families may indicate a novel association as a cancer susceptibility trait. PMID- 11857362 TI - Associations between family history of colorectal cancer and genetic alterations in tumors. AB - A family history of colorectal cancer has been consistently associated with an increased risk of developing colon cancer. However, there is limited information on the association between family history of colorectal cancer and genetic alterations that occur in colon tumors. In this study, we evaluate the association among genetic alterations of Ki-ras and p53, microsatellite instability and having a family history of colorectal cancer in a study of incident colon cancer cases (n = 1993) and population-based controls (n = 2,410). Although there was a slight nonsignificant increase in risk of having an unstable tumor among those with a family history of colorectal cancer, this increase in risk disappeared after excluding those people with a known mutation in either of the mismatch repair genes hMLH1 or hMSH2. A family history of colorectal cancer was not associated with Ki-ras mutations overall, although those with a G to T mutation of the second base of codon 12 were more likely to have a family history of colorectal cancer than were those without this specific type of Ki-ras mutation. Cases with p53 mutations were less likely to have a family history of colorectal cancer than were cases without a p53 mutation. We believe that, given the general lack of association between having a family history of colorectal cancer and genetic alterations in tumors, these alterations are acquired through disease pathways that involve exposure from diet, lifestyle or other environmental factors. PMID- 11857363 TI - A breast cancer screening programme operating in a liberal health care system: the Luxembourg Mammography Programme, 1992-1997. AB - The national breast cancer screening programme in Luxembourg, the Mammography Programme (MP), was launched in 1992. Its primary goal was to set up an organised breast cancer screening with biennial invitation-reinvitation of women 50-64 years, 2-view mammography, technical quality assurance, double-reading procedures and close monitoring of performance. In 1994, the decision was taken by health authorities to only reimburse screening mammograms done within the MP. We evaluated the performance of 3 screening rounds that took place in 1992-1997. In 1992, 4,815 women had opportunistic screening and 4,790 attended the MP. In 1997, 861 women had opportunistic screening and 8,603 attended the MP. In 1997, the participation rate in the MP reached 50%. The referral rate in the MP was 10% for initial screening and 5% for subsequent screening. Echographic examinations accounted for 71% of referrals. Per 1,000 screened women, biopsy and cancer rates were 18.0 and 8.0 for initial screening and 10.3 and 5.8 for subsequent screening. Twenty-one percent of the breast cancers diagnosed in screened women were interval cancers. The age-adjusted proportion of tumours >20 mm was 56.1% before 1992, 45.1% for women not (yet) screened by the MP, 27.0% for interval cancers, 26.6% for initial screening and 12.0% for subsequent screenings. Tumour size distribution was similar to that observed in the Dutch Breast Cancer Screening Programme, 1990-1993, except that in the latter programme, no difference was observed between initial and subsequent screening. The Luxembourg experience shows that in a liberal health care system, a policy of organising the screening process by reimbursing only those mammograms done within the context of the organised screening programme can drastically reduce opportunistic screening. Further efforts in the Mammography Programme will aim at increasing participation and look at ways to decrease referrals to echography. PMID- 11857364 TI - Height, weight, menstrual and reproductive factors and risk of gastric cancer among Japanese postmenopausal women: analysis by subsite and histologic subtype. AB - To clarify whether reproductive factors have an impact on gastric cancer in Japanese females, a case-control study was conducted using data from the Hospital based Epidemiologic Research Program at Aichi Cancer Center (HERPACC), Japan. The study subjects included 365 postmenopausal women with gastric cancer and 1,825 age-class frequency-matched noncancer outpatients presenting at Aichi Cancer Center in 1988-1998. Cases were further divided with regard to the anatomic subsite (upper third, middle third, lower third) and histologic subtypes (differentiated, nondifferentiated) and the association was evaluated using odds ratios (ORs) estimated by the logistic regression model, adjusting for potential confounding factors. A high body weight and corresponding body mass index at age 20 moderately increased the risk of gastric cancer, especially for middle third and nondifferentiated cancers. Risk fluctuation with early or late age at menarche and menopause and total duration of fertility was not consistent. Individuals with a high age at first parity tended to show decreased risk of cancer, irrespective of their subsite or histologic subtype. The ORs were decreased with a short average period of breastfeeding, especially for upper third and nondifferentiated cancers. From these results, however, it appears that height, weight, menstrual and reproductive factors have less impact on gastric cancer than environmental factors such as smoking and dietary habits or family history of gastric cancer. PMID- 11857365 TI - Antiangiogenic activity of endostatin inhibits C6 glioma growth. AB - Angiogenesis is a vital component of the development and progression of many human solid tumors. Glioblastoma multiforme is one of the most highly vascularised class of solid tumors. Thus, we have investigated the potential antitumourigenic activity of endostatin, an angiogenic inhibitor, in the rat C6 glioma model. We have engineered C6 cells that endogenously express mouse endostatin in order to assess the growth of C6 tumors in vivo when endostatin is constitutively expressed. Endostatin secreted by stably transfected C6 cells is biologically active as shown by its inhibition (26%) of bFGF-stimulated proliferation of BAECs in culture. The subcutaneous implantation of endostatin-C6 cells in athymic (nu/nu) mice resulted in a reduced tumor growth rate (90% inhibition) compared to control cell lines throughout the duration of our experiments. Tumor inhibition was associated with a 50% reduction in the number of vessels, which were also smaller in morphology. However, endostatin-C6 tumors were no more necrotic than control tumors. The implantation of endostatin-C6 cells into immunocompetent Wistar rat brains also resulted in reduced tumor volumes (71% inhibition) compared to controls. Tumor cells were sparsely localised along the injection tract but had not formed discrete tumors. Despite the inhibitory response mediated by endostatin on C6 growth, complete tumor inhibition or dormancy was not observed in either the athymic or immunocompetent tumor models. These findings demonstrate that the endogenous expression of endostatin by C6 glioma cells results in a reduced tumor growth rate in vivo that is associated with an inhibition of tumor angiogenesis. Our data suggest that endostatin should be developed as an adjuvant gene therapy for the effective treatment of gliomas. PMID- 11857366 TI - Intratumoral immunotoxin treatment of human malignant brain tumors in immunodeficient animals. AB - Treatment of malignant brain tumors remains a clinical challenge. New treatment modalities are under investigation and among these are intratumoral infusion of immunotoxins that bind to specific cell surface molecules on the malignant cells. We have compared the efficacy of the 425.3-PE immunotoxin (which targets the epidermal growth factor [EGF] receptor) with the well-known immunotoxin Tfn CRM107 (which targets the transferrin receptor), for the treatment of subcutaneous and intracranial human gliomas in nude animals. Bolus intratumoral administration of 1 microg Tfn-CRM107 or 425.3-PE into sc U87Mg tumors in nude mice reduced the tumor volume to 29 and 79%, respectively, of that in the control group 18 days after start of treatment. Higher doses of Tfn-CRM107 were toxic to the animals, whereas 425.3-PE was tolerated, with a dose-response relationship of up to 8 microg, a dose that reduced the tumor volume to 2% of control. In nude rats, treatment of intracerebral U87Mg tumors with Tfn-CRM107 proved ineffective and doses above 10 ng/animal were toxic to tumor-bearing rats. In contrast, intratumoral administration of 4 microg 425.3-PE increased symptom-free survival from 23 days to 40 days, with 2/9 surviving more than 90 days. We have recently shown that immunodeficient rats inoculated intracerebrally with precultured glioblastoma biopsy specimens develop highly infiltrative brain tumors. Direct interstitial infusion of immunotoxins into such tumors reduced the number of animals with detectable tumors at autopsy after 3 months, from 8/9 in the control animals to 4/6 and 2/6 in animals treated with Tfn-CRM107 and 425.3-PE, respectively. In conclusion, the anti-EGF receptor immunotoxin 425.3-PE exhibited promising efficacy, comparable to or better than that of Tfn-CRM107, an immunotoxin that in early clinical trials has been found to give responses in patients with brain tumors. PMID- 11857367 TI - Paclitaxel combined with fractionated radiation in vitro: a study with vulvar squamous cell carcinoma cell lines. AB - Concurrent paclitaxel and radiation has given promising results in the treatment of a variety of solid tumors. We wanted to test the efficacy of this combination for vulvar carcinoma, which currently has a poor outcome in advanced stages. The radiation sensitivity, sublethal damage repair (SLDR) capacity and effect of paclitaxel during fractionated radiation were assessed in our study on 7 vulvar inherently radioresistant squamous cell carcinoma (SCC) cell lines. The 96-well plate clonogenic assay was used. Survival data were fitted to the linear quadratic model. The area under the curve (AUC), equivalent to mean inactivation dose (D), was obtained with numerical integration. AUC ratios between single-dose radiation and fractionated radiation with or without paclitaxel were used to determine the SLDR of the cell lines and the effect of paclitaxel on it. Seven currently tested vulvar SCC cell lines were found to have a limited capacity of repairing sublethal damage (SLD). Only 3 of them presented SLDR of significance. The effect of concurrent radiation and paclitaxel was clearly additive when the radiation dose was fractionated in most of the cell lines. In addition, 2 of the cell lines having SLDR exhibited a trend toward losing the repair capacity when paclitaxel was present during the irradiation. In addition, the survival curve of the UM-SCV-1A cell line gave the impression of a true paclitaxel effect on SLDR. Paclitaxel used concurrently with fractionated radiation showed effectiveness on vulvar carcinoma. The effect was at least additive and could even be expected to abrogate the SLDR during split-dose radiation. PMID- 11857368 TI - Overexpression of BclXL in a human ovarian carcinoma cell line: paradoxic effects on chemosensitivity in vitro versus in vivo. AB - The effect of overexpressing the antiapoptotic protein BclXL in a human ovarian carcinoma cell line has been investigated in terms of sensitivity to the 2 major drugs used to treat this disease, paclitaxel and cisplatin. Stable transfection of BclXL into CH1 cells, which are relatively sensitive to cisplatin, resulted in around 2.7-fold higher expression in comparison with empty vector controls. However, this level of overexpression did not result in significant resistance in vitro to paclitaxel or cisplatin at the 50% inhibition level, using either short term (4-day) growth inhibition or longer term colony-forming assays. By contrast, parallel subcutaneous xenograft models of these isogenic ovarian carcinoma cells in vivo, differing only in BclXL status, showed that this low-level BclXL overexpression conferred significant resistance to both paclitaxel and cisplatin in comparison with parent, nontransfected tumours. Whereas parent non-BclXL transfected tumours were highly responsive, with the disappearance of tumours for at least 50 days post treatment, tumours overexpressing BclXL grew back after 30 and 20 days after treatment with paclitaxel and cisplatin, respectively. These differences in responsiveness to paclitaxel in vivo were not attributable to any significant changes in the delivery of drug to the tumour. These data suggest that the responsiveness of ovarian cancer to paclitaxel and cisplatin in vivo, and therefore perhaps clinically, is influenced by levels of the antiapoptotic protein BclXL. Such effects may be missed in vitro when using short-term growth inhibition or clonogenic assays. PMID- 11857369 TI - Dietary intake of folic acid and colorectal cancer risk in a cohort of women. AB - Folate is crucial for normal DNA methylation, synthesis and repair, and deficiency of this nutrient is hypothesized to lead to cancer through disruption of these processes. There is some evidence to suggest that relatively high dietary folate intake might be associated with reduced colorectal cancer risk, especially among individuals with low methionine intake. A case-cohort analysis was undertaken within the cohort of 56,837 women who were enrolled in the Canadian National Breast Screening Study and who completed a self-administered dietary questionnaire. During follow-up to the end of 1993, a total of 389 women were diagnosed with colorectal cancer, identified by linkage to the Canadian Cancer Database. For comparative purposes, a subcohort of 5,681 women was randomly selected from the full dietary cohort at baseline. After exclusions for various reasons, the analyses were based on 295 cases and 5,334 non-cases. Folate intake was inversely associated with colorectal cancer risk (IRR = 0.6, 95% CI = 0.4-1.1, p for trend = 0.25). The inverse association was essentially similar among individuals with low and high methionine intake, and was similar for colon and rectal cancers when those endpoints were analyzed separately. Among individuals with low methionine intake, folate intake did not appear to lower the risk of rectal cancer, a finding that may be due, in part, to the low number of cases in the subgroup analysis. Overall, our data lend some support to the hypothesis that high folate intake is associated with a reduced risk of colorectal cancer. PMID- 11857370 TI - Sequence variation and physical state of human papillomavirus type 16 cervical cancer isolates from Australia and New Caledonia. AB - Sequence diversity over 2600 nucleotides of the upstream regulatory region (URR) and the E6 and E2/E4 genes of 34 human papillomavirus (HPV)16 cervical cancer isolates from Australia and New Caledonia was investigated. One 81 base duplication, 41 single base substitutions and 1 single base insertion were identified in the URRs. Some of these changes are reported here for the first time. Several of the 19 changes impacting transcription factor binding sites had the potential to alter promoter activity. Twenty-eight (82%) of the variants belonged to the European lineage, 4 (12%) were Asian and 2 (6%) were Asian American. Eighteen of 27 (67%) isolates where the E6 gene was examined contained amino acid substitutions. Of 13 isolates sequenced with intact E2 genes, 12 (92%) contained amino acid substitutions in the E2 protein and 3 (23%) amino acid substitutions in the overlapping E4 protein. Some of the changes in E6 and E2 may alter immunological epitopes or protein function. The physical state of HPV DNA was assessed by Southern hybridization and PCR for an intact E2 gene. Overall, 11 of 25 isolates contained only integrated HPV DNA, 10 only episomal HPV DNA and 4 both integrated and episomal DNA. No particular patterns of variation in the URR, E6 or E2/E4 genes predicted physical state. This investigation represents one of the most comprehensive studies of its kind and fills an important gap in global sequence data. PMID- 11857371 TI - Absence of SDHD mutations in primary nasopharyngeal carcinomas. PMID- 11857376 TI - Neurotrophin receptor p75(NTR) suppresses growth and nerve growth factor-mediated metastasis of human prostate cancer cells. AB - The loss of tumor- and/or metastasis-suppressor gene function contributes to the transformation of human prostate epithelial cells to a malignant pathology. Such a putative tumor-suppressor and metastasis-suppressor gene(s) has been mapped to the region of 17q21, which coincidentally is in the vicinity of the human gene locus for the neurotrophin receptor p75(NTR). The p75(NTR) is expressed in normal human prostate epithelial cells and exhibits an inverse association of p75(NTR) expression with the malignant progression of the prostate, consistent with a pathologic role of the p75(NTR) as a putative tumor and metastasis suppressor. Utilizing stable transfectants of the TSU-pr1 and PC-3 human prostate tumor cell lines that exhibit a rank order (dose-dependent) increase in p75(NTR) protein expression, we investigated the effects of the p75(NTR) in combination with its predominant ligand, nerve growth factor (NGF), on tumor cell growth. A rank order (dose-dependent) increase in p75(NTR) expression was found to suppress the growth of prostate tumors in severe combined immunodeficient (SCID) mice. Treatment of these tumors with NGF stimulated both proliferation as indicated by PCNA expression and apoptosis as indicated by TUNEL assay, the net result of which was no change in the overall growth of the tumors. However, NGF was found to increase the formation of satellite tumors, both contiguous and noncontiguous with respect to the primary tumor mass, indicating dose-dependent induction of metastasis. Significantly, the formation of satellite tumors was suppressed by the expression of p75(NTR). This suggests that p75(NTR) is a tumor suppressor of growth and a metastasis suppressor of NGF-stimulated migration of human prostate tumor cells. PMID- 11857377 TI - Concurrent overexpression of Ets-1 and c-Met correlates with a phenotype of high cellular motility in human esophageal cancer. AB - Hepatocyte growth factor (HGF) stimulates cell motility as well as mitotic activity of cells. High concentrations of HGF or overexpression of its cellular receptor c-Met in cancer have been reported. We analyzed the expression status of c-Met immunohistochemically in 76 cases of human esophageal cancer. Overexpression of c-Met was noted at a considerably high frequency. Intriguingly, c-Met overexpression was frequent in a specific type of cell nest formation in tumors, i.e., the small nest type, in which tumors form small, dispersed cell nests. Further immunohistochemical analyses using serial sections revealed a striking coincidence between overexpression of c-Met and its transcriptional factor, Ets-1. Overexpression of c-Met and Ets-1 was statistically more frequent in small nest type tumors. The close correlation in expression status between Ets 1 and c-Met was also confirmed using 6 established human esophageal cancer cell lines. In addition, cells that expressed high levels of Ets-1 and c-Met exhibited an extremely motile phenotype by HGF stimulation in vitro. The presence of HGF in tissue sections was confirmed using similar immunohistochemical approaches. These observations suggest that in human esophageal cancer cells the transcriptional factor Ets-1 upregulates the expression of c-Met and, consequently, confers on cells a highly motile phenotype leading to a specific form of tumor development. PMID- 11857378 TI - Significance of vascular endothelial growth factor (VEGF)/soluble VEGF receptor-1 relationship in breast cancer. AB - Angiogenesis, the formation of new blood vessels, is controlled by a balance between positive and negative endothelial regulatory factors. Soluble vascular endothelial growth factor receptor-1 (sVEGFR1), a naturally occurring soluble form of VEGFR1, is a negative counterpart of the vascular endothelial growth factor (VEGF) signaling pathway, which has been characterized as one of the most important endothelial regulators in human tumor angiogenesis. In our study, we examined the expression of sVEGFR1 in 110 primary breast carcinomas, and assessed its clinical significance. Ninety-four of 110 tumors showed > or = 0.1 ng/mg protein of sVEGFR1 (range:0. 1-6.9 ng/mg protein; median: 1.03 ng/mg protein) as determined by a specific enzyme-linked immunosorbent assay (ELISA). Immunoblot analysis confirmed the presence of sVEGFR1 in breast tumor tissues. The levels of sVEGFR1 were correlated significantly with the levels of VEGF. There was no significant correlation between the levels of sVEGFR1 and any clinico pathological factors including age, menopause, nodal involvement and hormone receptor status. A univariate prognosis analysis showed that the intratumoral VEGF status, as determined by ELISA, was a significant prognostic indicator, but sVEGFR1 status was not. In the combined analysis, however, the ratio of sVEGFR1 and VEGF levels provided more statistically significant prognostic value than VEGF status alone. Tumors in which the sVEGFR1 levels exceeded VEGF levels 10 fold had a markedly favorable prognosis. Multivariate analysis also demonstrated that the ratio of sVEGFR1 and VEGF was an independent prognostic indicator after nodal status. In conclusion, sVEGFR1, an intrinsic inhibitor of VEGF, frequently co-expressed with VEGF in primary breast cancer tissues. The intratumoral balance between sVEGFR1 and VEGF levels might be crucial for the progression of breast cancer. PMID- 11857379 TI - Overexpression of fatty acid synthase is an early and common event in the development of prostate cancer. AB - The expression of fatty acid synthase (FAS), a key lipogenic enzyme and potential target for antineoplastic therapy, was analyzed in 87 frozen needle biopsies of prostate cancer using a highly sensitive immunohistochemical detection technique (Envision). In comparison to normal or benign, hyperplastic glandular structures, which were all negative for FAS staining, immunohistochemical signal was evident in 24/25 low grade prostatic epithelial neoplasia (PIN) lesions, in 26/26 high grade PIN lesions and in 82/87 invasive carcinomas. Staining intensity tended to increase from low grade to high grade PIN to invasive carcinoma. Cancers with a high FAS expression had an overall high proliferative index. No correlation was found between FAS expression and lipid accumulation. These findings indicate that increased FAS expression is one of the earliest and most common events in the development of prostate cancer, suggesting that FAS may be used as a general prostate cancer marker and that antineoplastic therapy based on FAS inhibition may be an option for chemoprevention or curative treatment for nearly all prostate cancers. PMID- 11857380 TI - Expression of membrane type 1 matrix metalloproteinase (MT1-MMP) in A2058 melanoma cells is associated with MMP-2 activation and increased tumor growth and vascularization. AB - Membrane-type metalloproteinase-1 (MT1-MMP) is a transmembrane metalloproteinase overexpressed in tumors, which plays a major role in the first step of pro-MMP-2 activation, leading to the generation of an intermediate 62 kDa species. The second step of MMP-2 activation that yields to the mature form is less understood and could involve an autocatalytic process and/or the activity of the plasminogen/plasmin system. Human melanoma A2058 cells, which express MMP-2 only in its pro-form, were used to determine the role of MT1-MMP during pericellular proteolysis and tumor progression. The induction of MT1-MMP overexpression by MT1 MMP cDNA transfection initiated the first step of MMP-2 activation. We provide evidence that a cooperation between the plasminogen/plasmin system and MT1-MMP endowed the cells with the ability to fully activate MMP-2 and with enhanced invasive properties in vitro. When injected subcutaneously in nude mice, MT1-MMP expressing clones induced rapid tumor growth and high tumor vascularization, while the control clones were poorly or not tumorigenic. Our data provide the first demonstration, in an experimental model, that MT1-MMP expression by tumor cells promotes tumor vascularization. PMID- 11857381 TI - Identification of bradykinin receptors in clinical cancer specimens and murine tumor tissues. AB - Bradykinin (BK) has multiple pathophysiologic functions such as induction of vascular permeability and mitogenesis, and it triggers the release of other mediators such as nitric oxide in inflammatory and cancer tissues. To explore the pathophysiologic roles of BK in tumor, we examined the distribution of BK B2 receptors in human adenocarcinoma (lung, stomach), lymphoma (lymph node), hepatoma, squamous cell carcinoma (lung) and carcinoid (duodenum), and in mouse colon adenocarcinoma 38 (C-38) and sarcoma 180 (S-180) tumor tissues. Immunohistochemical staining of tumor tissues with an anti-BK B2 receptor antibody, or autoradiography with the B2 receptor antagonist [125I]HOE 140 (D-Arg [Hyp Thi D-Tic Oic8]-BK) and the B2 receptor agonist [3H]BK indicated the presence of B2 receptors in all human tumor cells and murine S-180 and C-38 cells. Specific binding of [3H]HOE 140 was observed in S-180 cells with a Kd of 2.1 nM. Binding of [125I]HOE 140 to S-180 cells was competed by an excess amount (20-100 times) of nonradiolabeled HOE 140 or BK, but not by BK B1 receptor agonist des-Arg9-BK. These results provide direct evidence that the BK B2 receptor is expressed in human cancer and experimental murine tumors, which suggests a potential role for BK in inducing pathologic signal transduction in cancer growth and progression, nitric oxide production and vascular permeability enhancement in tumors. BK antagonists may thus have applications in the modulation of cancer growth and in paraneoplastic syndromes. PMID- 11857382 TI - Topotecan inhibits VEGF- and bFGF-induced vascular endothelial cell migration via downregulation of the PI3K-Akt signaling pathway. AB - Angiogenesis plays a crucial role in tumor growth and metastases. The extent of angiogenesis correlates with the increased invasion and metastasis in a variety of human neoplasms. Vascular endothelial cell proliferation and migration are critical steps in angiogenesis and are regulated by various growth factors, such as vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF). The topoisomerase I inhibitor topotecan (10-hydroxy-9-dimethylaminomethyl (S)-camptothecin) is a water-soluble camptothecin analogue and possesses an indirect in vivo antitumor effect mediated through the inhibition of angiogenesis. We found that topotecan inhibited VEGF- and bFGF-induced migration of human umbilical vein endothelial cells (HUVECs) in vitro. The migration of HUVECs was also inhibited by a phosphatidylinositol 3-kinase (PI3K) inhibitor, LY294002. Thus, we investigated the possibility that topotecan's antiangiogenic property might be mediated by its inhibitory effect on VEGF- and bFGF-induced activation of the PI3K-Akt signaling pathway. We found that topotecan treatment decreased the amount of the phosphorylated (activated) form of Akt, but not the amount of Akt protein, in HUVECs. Moreover, transient transfection of constitutive active akt cDNA into HUVECs reversed the topotecan-mediated decrease in HUVEC migration. These results suggest that the antiangiogenic activity of topotecan is mediated in part by downregulating the PI3K-Akt signaling pathway. PMID- 11857383 TI - Prevalence of the BRCA2 6174 del T mutation in Israeli uveal melanoma patients. AB - Substantial differences exist in the incidence rates of uveal melanoma (UM) among Israeli Jewish subpopulations: high in immigrants from North America and Europe (Ashkenazic) and low in immigrants from Africa and Asia (Sepharadic). This trend persists in Israeli-born individuals when stratified by their ancestral place of birth. There have been several anecdotal reports of uveal melanoma occurring in breast cancer families with mutations in the BRCA2 gene as well as one systematic study reporting BRCA2 mutations in UM. A single BRCA2 mutation, 6174 del T, occurs in about 1% of the Ashkenazic population and rarely in non-Ashkenazic. To assess the contribution of this germline mutation to uveal melanoma in Jewish Israeli patients, we tested this relationship through analysis of blood samples from a series of UM patients. A total of 153 cases (84 female, 69 male) were available for study, which represents 30% of all cases of UM diagnosed in Israel during the period 1984-1999 (82% for the period 1992-1999). Of the 143 UM patients for which a result could be obtained (4 due to refusals, 6 due to damage to the blood sample), 4 (2.8%, 95% confidence interval [CI] 0-5.6) carried the 6174 del T mutation. Assuming a population frequency of the mutation of 1% as estimated among Ashkenazic Jews in the United States, the probability of observing 4 or more carriers with the 6174 del T mutation, assuming no relationship between uveal melanoma and BRCA2, is 0.057. Although our study confirms the relationship between uveal melanoma and BRCA2, it is clear that the 6174 del T mutation accounts for only a small fraction of all Israeli UM cases. Therefore, BRCA2 mutations are likely to account for an even smaller proportion in populations with low frequencies of BRCA2 alterations. PMID- 11857384 TI - Detection of peptide-specific cytotoxic T-lymphocyte precursors used for specific immunotherapy of pancreatic cancer. AB - The prognosis of pancreatic cancer is extremely poor with a 5-year survival of approximately 3%. Thus, the development of new treatment modalities, including a specific immunotherapy, is required. Our study investigated whether cytotoxic T lymphocyte (CTL) precursors reacting to peptides with vaccine candidates (13 peptides for HLA-A2+ or -A24+ patients, respectively) were detectable in the prevaccination peripheral blood mononuclear cells (PBMCs) of 15 pancreatic cancer patients. Peptide-specific CTL precursors were detectable in the majority (11 of 15, 73%) of patients, with a mean positive number of 1.5 peptides (ranging from 0 5 peptides) per patient. Positive peptide profiles varied among patients. These results may provide a scientific basis for a new kind of cancer immunotherapy, namely, a CTL precursor-oriented peptide vaccine, for pancreatic cancer patients. PMID- 11857385 TI - Impact of human bladder cancer cell architecture on autologous T-lymphocyte activation. AB - To investigate the influence of tumor cell architecture on T-cell activation, we used an autologous human model based on 2 bladder tumor cell lines as targets for cytotoxic tumor-infiltrating lymphocytes (TILs). These tumor cell lines were grown in vitro as either standard 2-dimensional (2D) monolayers or 3-dimensional (3D) spheroids. T-cell activation was determined by measuring the production of three major cytokines (tumor necrosis factor, granulocyte/macrophage colony stimulating factor and interferon-gamma), known to be secreted by most activated TILs. Changes in the architecture of target cells from 2D to 3D induced a dramatic decrease in their capacity for stimulating TILs. Interestingly, neither TIL infiltration nor MHC class I, B7.1 costimulatory or lymphocyte function associated factor-3 adhesion molecule downregulation played a major role in this decrease. These findings demonstrate that tumor architecture has a major impact on T-cell activation and might be implicated in the escape of tumor cells from the immune system. PMID- 11857386 TI - Tumor-specific CD8+ T lymphocytes derived from the peripheral blood of prostate cancer patients by in vitro stimulation with autologous tumor cell lines. AB - To identify tumor-associated antigens as putative targets for developing immunotherapies against prostate cancer, we investigated the ability of T cells derived from the peripheral blood lymphocytes of prostate cancer patients to recognize autologous tumor cells. The technical challenge of growing in vitro carcinoma cell lines from small prostate cancer samples was previously addressed by immortalization of early epithelial cell cultures with the HPV16 transforming proteins E6 and E7 and by genetic characterization of the carcinoma and normal prostate cell lines. In our study, peripheral blood lymphocytes were stimulated in vitro using autologous IFNgamma-treated prostate carcinoma cells transduced with the B7.1 molecule as a source of T-cell costimulation. Tumor-specific CD8+ T lymphocytes were obtained from 3 of 6 prostate cancer patients tested and included T cells restricted by classical and nonclassical HLA molecules. In 1 case, we demonstrated that the prostate cancer-reactive T cells were TCRalpha/beta+ and recognized autologous tumor cells but not autologous normal cells in the context of HLA-B or -C molecules. These results validate the approach of in vitro stimulation of peripheral blood lymphocytes from prostate cancer patients with autologous tumor cell lines to isolate prostate cancer specific T cells and demonstrate the existence of a functionally diverse immune response against prostate cancer. PMID- 11857387 TI - A novel approach for modifying tumor cell-derived plasma membrane vesicles to contain encapsulated IL-2 and engrafted costimulatory molecules for use in tumor immunotherapy. AB - The genetic modification of tumor cells and delivery of cytokines have been proposed as useful strategies in the development of anti-tumor vaccines; however, a number of factors limit their use in clinical settings. To facilitate vaccine development, we explored the possibility of modifying plasma membrane vesicles (PMV) by using a novel chelator lipid, nitrilotriacetic acid ditetradecylamine (NTA-DTDA). Our analyses by flow cytometry show that NTA-DTDA can be incorporated into PMV prepared from murine P815 mastocytoma and that the incorporated NTA-DTDA permits anchoring or "engraftment" onto the vesicle surface of hexahistidine tagged proteins such as recombinant forms of the costimulatory molecules B7.1 and CD40. The engrafted PMV also can incorporate and deliver the immunostimulatory cytokine Interleukin-2 (IL-2). Our results show that modified PMV derived from P815 cells bind the murine T cell clone D10 in a receptor-ligand dependent manner, inducing cell adhesion and promoting cell survival in vitro. The modified PMV can bind syngeneic T cells, stimulating T cell proliferation and cytotoxic T cell responses. Moreover, when used as vaccines in syngeneic animals, the modified vesicles induce significant protection against challenge with the native P815 tumor. The results indicate that PMV modified by engraftment of recombinant forms of B7.1 and CD40 and incorporation of IL-2 can be used to modulate immune responses, which provides a novel approach for the development of anti-tumor vaccines and cancer immunotherapies. PMID- 11857388 TI - Analysis of the antibody repertoire of astrocytoma patients against antigens expressed by gliomas. AB - The molecular characterization of antigens preferentially or exclusively expressed by astrocytomas and recognized by the autologous immune system are a prerequisite for the development of specific vaccines. To identify such antigens, we screened 5 cDNA expression libraries derived from astrocytomas and other gliomas for reactivity with high-titered IgG antibodies in the sera of astrocytoma patients using SEREX, the serologic identification of antigens by recombinant cDNA expression cloning. Autologous and allogeneic SEREX analysis of >5 x 10(6) clones with the sera of 18 astrocytoma patients revealed 10 antigens: the differentiation antigen glial fibrillary acidic protein (GFAP), Bax-inhibitor 1 (which was overexpressed in all glioma samples tested), 3 other molecules involved in the regulation of gene expression and proliferation (the nm23-H2 encoded nucleoside diphosphate kinase B, the Ran binding protein-2 and a DNA binding protein encoded by the son gene), SP40,40 (a complement inhibitory molecule), the chaperonin TCP-1, calnexin and 2 new gene products. No immune responses were detected against the "shared tumor" or "cancer testis antigens" that are regularly expressed in gliomas. Antibody responses in astrocytoma patients against antigens expressed by gliomas were rare and, with the exception of Bax-inhibitor 1 and the product of the son gene, were also found in apparently healthy controls. We conclude that although astrocytomas express a broad spectrum of antigens, they elicit antibody responses only rarely, most likely because of their intrinsic immunosuppressive effects. PMID- 11857389 TI - N-3 and N-6 fatty acids in breast adipose tissue and relative risk of breast cancer in a case-control study in Tours, France. AB - Experimental studies have indicated that n-3 fatty acids, including alpha linolenic acid (18:3 n-3) and long-chain n-3 polyunsaturated fatty acids inhibit mammary tumor growth and metastasis. Earlier epidemiological studies have given inconclusive results about a potential protective effect of dietary n-3 polyunsaturated fatty acids on breast cancer risk, possibly because of methodological issues inherent to nutritional epidemiology. To evaluate the hypothesis that n-3 fatty acids protect against breast cancer, we examined the fatty acid composition in adipose tissue from 241 patients with invasive, nonmetastatic breast carcinoma and from 88 patients with benign breast disease, in a case-control study in Tours, central France. Fatty acid composition in breast adipose tissue was used as a qualitative biomarker of past dietary intake of fatty acids. Biopsies of adipose tissue were obtained at the time of surgery. Individual fatty acids were measured as a percentage of total fatty acids, using capillary gas chromatography. Unconditional logistic regression modeling was used to obtain odds ratio estimates while adjusting for age, height, menopausal status and body mass index. We found inverse associations between breast cancer-risk and n-3 fatty acid levels in breast adipose tissue. Women in the highest tertile of alpha-linolenic acid (18:3 n-3) had an odds ratio of 0.39 (95% confidence intervals [CI] = 0.19-0.78) compared to women in the lowest tertile (trend p = 0.01). In a similar way, women in the highest tertile of docosahexaenoic acid (22:6 n-3) had an odds ratio of 0.31 (95% CI = 0.13-0.75) compared to women in the lowest tertile (trend p = 0.016). Women in the highest tertile of the long chain n-3/total n-6 ratio had an odds ratio of 0.33 (95% confidence interval = 0.17-0.66) compared to women in the lowest tertile (trend p = 0.0002). In conclusion, our data based on fatty acids levels in breast adipose tissue suggest a protective effect of n-3 fatty acids on breast cancer risk and support the hypothesis that the balance between n-3 and n-6 fatty acids plays a role in breast cancer. PMID- 11857390 TI - Quantitative evaluation of oxidative stress, chronic inflammatory indices and leptin in cancer patients: correlation with stage and performance status. AB - In advanced cancer patients, the oxidative stress could take place either at the onset of disease or as a function of disease progression. To test this hypothesis, the following parameters were investigated: the erythrocyte activity of the enzymes superoxide dismutase (SOD) and glutathione peroxidase (GPx), the serum activity of glutathione reductase (GR) and the serum total antioxidant status (TAS). The total antioxidant capacity of plasma LMWA was evaluated by the cyclic voltammetry methodology. We further determined the serum levels of proinflammatory cytokines (IL-6 and TNFalpha), IL-2, leptin and C-reactive protein (CRP). All of these parameters have been correlated with the most important clinical indices of patients such as Stage of disease, ECOG PS and clinical response. Eighty-two advanced stage cancer patients and 36 healthy individuals used as controls were included in the study. Our findings show that SOD activity was significantly higher in cancer patients than in controls and GPx activity was significantly lower in cancer patients than in controls. Serum values of IL-6, TNFalpha and CRP were significantly higher in patients than in controls. Serum leptin values of cancer patients were significantly lower than controls. SOD activity increased significantly from Stage II/ECOG 0-1 to Stage IV/ECOG 0-1, whereas it decreased significantly in Stage IV/ECOG 3. GPx activity decreased significantly in Stage IV/ECOG 2-3. An inverse correlation between ECOG PS and serum leptin levels was found. Serum levels of IL-2 decreased from Stage II/ECOG 0-1 to Stage IV/ECOG 2-3. A direct correlation between Stage/ECOG PS and serum levels of both IL-6 and CRP was observed. Cisplatin administration induced a significant increase of GPx after 24 hr. In conclusion, this is the first study that shows that several "biological" parameters of cancer patients such as antioxidant enzyme activity, cytokines, leptin and CRP strictly correlate with the most important clinical parameters of disease such as Stage and ECOG PS. PMID- 11857391 TI - Melanoma in adolescents: a case-control study of risk factors in Queensland, Australia. AB - The incidence of melanoma increases markedly in the second decade of life but almost nothing is known of the causes of melanoma in this age group. We report on the first population-based case-control study of risk factors for melanoma in adolescents (15-19 years). Data were collected through personal interviews with cases, controls and parents. A single examiner conducted full-body nevus counts and blood samples were collected from cases for analysis of the CDKN2A melanoma predisposition gene. A total of 201 (80%) of the 250 adolescents with melanoma diagnosed between 1987 and 1994 and registered with the Queensland Cancer Registry and 205 (79%) of 258 age-, gender- and location-matched controls who were contacted agreed to participate. The strongest risk factor associated with melanoma in adolescents in a multivariate model was the presence of more than 100 nevi 2 mm or more in diameter (odds ratio [OR] = 46.5, 95% confidence interval [CI] = 11.4-190.8). Other risk factors were red hair (OR = 5.4, 95%CI = 1.0 28.4); blue eyes (OR = 4.5, 95%CI = 1.5-13.6); inability to tan after prolonged sun exposure (OR = 4.7, 95%CI = 0.9-24.6); heavy facial freckling (OR = 3.2, 95% CI = 0.9-12.3); and family history of melanoma (OR = 4.0, 95%CI = 0.8-18.9). Only 2 of 147 cases tested had germline variants or mutations in CDKN2A. There was no association with sunscreen use overall, however, never/rare use of sunscreen at home under the age of 5 years was associated with increased risk (OR = 2.2, 95%CI = 0.7-7.1). There was no difference between cases and controls in cumulative sun exposure in this high-exposure environment. Factors indicating genetic susceptibility to melanoma, in particular, the propensity to develop nevi and freckles, red hair, blue eyes, inability to tan and a family history of the disease are the primary determinants of melanoma among adolescents in this high solar radiation environment. Lack of association with reported sun exposure is consistent with the high genetic susceptibility in this group. PMID- 11857392 TI - TP53 mutation pattern of esophageal squamous cell carcinomas in a high risk area (Southern Brazil): role of life style factors. AB - In an attempt to correlate the TP53 mutation pattern of squamous cell carcinomas of the esophagus (ESCC) and life style factors of patients from the high risk area Rio Grande do Sul, Brazil, 135 ESCC were analyzed, after prescreening by p53 immunohistochemistry, by SSCP and DNA sequencing of TP53, exon 5-9. Forty-nine somatic TP53 mutations (and 1 case with p53 polymorphism) were identified as missense (n = 39), frameshift (n = 6), silent (n = 1), amber (n = 1) or intron border mutations (n = 2) that cause splicing aberrations. They were preferentially found in exon 5 (36.7%) and exon 8 (32.7%). Several mutations were located in the mutation hot spot codons 248, 273 and 282, mainly at CpG sites. Transition mutations were observed in 53.1% (among them 50% G > A), transversion mutations in 34.7% (among them 47.1% G > T) and frameshifts in 12.2%, the latter 2 mainly in smokers and alcohol drinkers. Transitions were more prevalent in females than in males (p < 0.05). TP53 mutations, mainly transversions, were more frequently found in heavy smokers (p = 0.03), with the same tendency after chronic alcohol consumption. Comparison with the worldwide IARC database disclosed differences in the TP53 mutation pattern of the Brazilian tumors, with a higher accumulation of TP53 mutations in exon 8 and a higher prevalence of transition mutations. Mutations at the reported hot spot codon 176 were missing. Although difficult because of the documented coexposure to various life style risk factors in most patients of this series, the hypothesis is proposed that besides smoking and alcohol drinking the commonly consumed hot mate tea in this high risk area for ESCC is responsible for this different pattern of TP53 mutations because of chronic hyperthermic irritation and inflammation in the esophagus with an endogenous formation of radicals or carcinogenic factors that lead to a higher prevalence of transition mutations. PMID- 11857393 TI - Familial and second esophageal cancers: a nation-wide epidemiologic study from Sweden. AB - A few case-control studies have been published on familial risks in esophageal cancer. Reliable data on familial risks are needed for prevention and clinical decisions. We used the nation-wide Swedish Family-Cancer Database on 10.1 million individuals and close to 6000 esophageal cancers to calculate standardized incidence ratios (SIRs) and 95% confidence intervals (CIs) for esophageal cancer in 0-66-year-old offspring by cancers in family members. Additionally, SIRs for second esophageal cancers were analyzed. The SIR for esophageal cancer was 3.91 (95% CI 1.55-7.35) when a parent presented with esophageal cancer and 4.91 (95% CI 1.77-9.62) when a parent presented with squamous cell carcinoma. The sibling risk for esophageal cancer was increased but was based on 1 pair only. The population-attributable proportion of familial esophageal cancer was 0.70%. Risks for second esophageal cancers were increased after upper aerodigestive tract, esophageal, stomach, larynx, and lung cancers. The data on second cancers suggest that environmental factors are important for esophageal cancer and probably contribute to the familial clustering. However, the high familial risk of 3.91 is unlikely without the involvement of heritable factors. The population attributable proportion of familial esophageal cancer is small. PMID- 11857394 TI - Social class and risk of Hodgkin's disease in young-adult women in 1988-94. AB - Hodgkin's disease (HD) risk in young adults has been associated with higher childhood social class. Although recent decades have witnessed increases in both young-adult HD incidence rates and the socioeconomic affluence reported to influence risk, social class risk factors have not been reexamined. For 204 cases and 254 controls aged 19-44 years from a population-based case-control study of HD diagnosed in 1988-94 in San Francisco area females, we evaluated social class predictors of HD overall and for subgroups defined by age and by ethnicity. HD was associated weakly with a few childhood social class markers but more strongly with combinations of these variables. Risk was higher for women with family-owned than rented childhood homes; for US-born women with single vs. shared bedrooms at age 11; and for women with 2+ births who were from smaller than larger childhood households. These patterns differed by age, with risk appearing to increase over the young-adult years for some factors and to decrease for others. In whites, risk was additionally associated with having a single childhood bedroom in larger households, and with tall adult height in women from smaller childhood households. In nonwhites, risk was higher for single bedrooms at age 11 in smaller childhood households, taller height and higher maternal education. Most study findings support the hypothesis that HD development in young adults follows protection from early exposure to other children. Variation in risk by age suggests differing etiologies across young adulthood, or the importance of birth cohort-appropriate social-class measures. Negative findings for previously reported risk factors may reflect their insufficient heterogeneity of exposure or their failure to measure cohort-relevant exposures in this population. PMID- 11857395 TI - Hormone receptors and proliferation in breast carcinomas of equivalent histologic grades in pre- and postmenopausal women. AB - Breast cancers in younger, premenopausal women are more likely to exhibit an adverse prognostic profile (including negative steroid hormone receptors and a high rate of cellular proliferation) and poor outcome than breast cancers in postmenopausal women. It has been hypothesized that this adverse prognostic profile is a result of the higher histologic grade of breast cancers in pre- compared with post-menopausal women. To assess the association of expression of steroid hormone receptors and indicators of proliferation while controlling for histologic grade, we identified 100 infiltrating ductal carcinomas from premenopausal women 45 years of age or younger and 100 from postmenopausal women 65 years of age or older. The carcinomas were selected so that the histologic grades (low versus high) were distributed equally between the 2 groups. Estrogen receptors (ER), progesterone receptors (PR), p27(Kip1) and Ki-67 (to measure rate of proliferation) were assessed by immunohistochemistry and compared between groups. Clinical information and survival data were also analyzed. ER content was lower and proliferation was higher in carcinomas in premenopausal women (p = 0.048 and p = 0.005, respectively). By univariate analysis, p27(Kip1) and PR were not different between the groups; however, in multivariate analysis, p27(Kip1) was higher in postmenopausal women, but only in a subgroup with highly proliferative carcinomas. Overall survival was similar in the pre- and postmenopausal women. Furthermore, low p27(Kip1) and African-American ethnicity predicted a poorer overall survival in the premenopausal, but not in the postmenopausal, women in our study. After controlling for histologic grade, a lower expression of ER and a higher proliferative index were detected in breast carcinomas in premenopausal women. Therefore, some prognostic indicators, such as ER and proliferative rate, may be more closely associated with menopausal status than histologic grade. Our data also suggest that some prognostic factors are not equally effective as predictors of survival in pre- and postmenopausal women. PMID- 11857396 TI - In vitro and in vivo downregulation of MRP1 by antisense oligonucleotides: a potential role in neuroblastoma therapy. AB - The expression of MRP1 (multidrug resistance protein-1) is associated with chemoresistance and poor prognosis in neuroblastoma. MRP1 antisense oligonucleotides were used in an in vivo mouse-human xenograft model of neuroblastoma to downregulate MRP1. The MRP1 ASO reduced protein levels of MRP1 to an average of 40% of the nil treated controls (p = 0.007). There was significant chemosensitisation to single-agent chemotherapy, VP16 (etoposide), at 1 microg/mL (p = 0.035) and 10 microg/mL (p = 0.02) in comparison to tumours not receiving oligonucleotides. In contrast, MDR1-ASO produced significant chemosensitisation only at 10 microg/mL of VP16 (p = 0.029). No significant chemosensitisation was seen following nonsense oligonucleotides. The downregulation of MRP1 was also associated with an increase in tumour cell death (79% increase in apoptosis index p = 0.0313) and a reduction in cell turnover (42% reduction in mitotic index p = 0.0313), which was not seen with any other oligonucleotide. This new and novel perspective of MRP1 function, which is an apparent involvement in apoptosis and cell cycle progression in neuroblasts, presents a fresh avenue for investigation of the biologic consequences of MRP1 expression that occurs in many tumour cell types. Our work is the first to concurrently explore the effects of downregulation of MRP1 and MDR1 by antisense oligonucleotides in a neuroblastoma xenograft model. It provides rationale for the investigation of therapy adjuvants such as antisense oligonucleotides in the treatment of this malignancy. PMID- 11857397 TI - Therapeutic efficacy of IL-2-loaded hydrogels in a mouse tumor model. AB - Interleukin-2 (IL-2) is a highly effective anticancer drug if it is applied locally for 5 consecutive days. In most cases this requires 5 invasive treatments, which is not usually acceptable for either the patient or the clinician. For this reason we have developed dextran-based hydrogels from which the required amount of encapsulated IL-2 (1-4 x 10(6) IU of IL-2) is gradually released during 5-10 days. Initially IL-2-containing macroscopic cylinder-shaped gels (implants), and later IL-2-containing injectable microspheres, were developed. These preparations were characterized in vitro, and the therapeutic activity was tested in DBA/2 mice with SL2 lymphosarcoma. The therapy was given to mice with a large and extensively metastasized tumor load (at least 5% of the body weight). If 1-4 x 10(6) IU of IL-2 was slowly released from the hydrogels over a period of 5-10 days, the therapeutic effects were very good and comparable to the effects of free IL-2 injections for 5 consecutive days. In conclusion, dextran-based hydrogels are promising systems for the controlled release of IL-2. PMID- 11857398 TI - A phase I-II study of docetaxel-ifosfamide-cisplatin (DIP) combination chemotherapy regimen in advanced nonsmall cell lung cancer. AB - In an attempt to develop more effective chemotherapy regimens in advanced nonsmall cell lung cancer (NSCLC), we evaluated docetaxel-ifosfamide-cisplatin (DIP) based on our previous experience with paclitaxel-ifosfamide-cisplatin. Patients with advanced NSCLC (stages III-IV), WHO-PS< or =2, no prior chemotherapy and unimpaired hematopoietic and organ function were eligible. Chemotherapy was administered in successive dose levels (DLs) and included docetaxel (80-100 mg/m2 day 1), ifosfamide (4-5 g/m2) and cisplatin (80-100 mg/m2), both divided over days 1 and 2 every 21 days. G-CSF (lenograstin) was administered from days 4-13. Fifty-five patients were accrued (phase I: 15; phase II: 40) and all are evaluable for response and toxicity: median age = 58 (40-72); PS = 1 (0-2); gender = 48 males, 7 females; stages IIIA = 8, IIIB = 19, IV = 28; and histologies were adenocarcinoma (29), squamous (20), large cell (6). Metastatic sites at diagnosis included lymph nodes (33), bone (8), liver (6), brain (6), lung nodules (9), adrenals (7) and soft tissue (1). The dose-limiting toxicity (DLT) was reached at DL4 (Docetaxel: 100 mg/m2-Ifosfamide: 5 g/m2 Cisplatin: 100 mg/m2) consisting of 2 cases of febrile neutropenia (FN), and DL3 (Docetaxel: 100 mg/m2-Ifosfamide: 5 g/m2-Cisplatin: 80 mg/m2) was considered as the maximum tolerated dose (MTD) and recommended for further phase II testing. Among evaluable patients in phase II, 31/46 (67%; CI = 54-81%) responded; 4 were complete responses, 27 partial responses, 12 with stable disease and 3 with progressive disease. The median response duration was 7 months (2-21+), median time to progression (TTP) 8 months (1-23+) and median overall survival (OS) 13 months (2-23+). The 1-year survival was 57%. Grade (Gr) 3/4 toxicities included neutropenia 39/46 with 27 developing Gr4 (< or =7 days) and 20% FN managed successfully with broad-spectrum antibiotics, thrombocytopenia Gr3 3/46-Gr4 1/46, no Gr3 neuropathy, Gr1-2 CNS toxicity in 12, no renal toxicity, 15 Gr2 myalgias, 17 Gr2 diarrhea and 10 Gr3 vomiting. In the present phase I-II study, DIP appears highly active and tolerable in advanced NSCLC in the outpatient setting. Randomized comparisons to current standard 2-drug regimens will be warranted. PMID- 11857399 TI - Dysregulation of phosphatidylinositol 3-kinase and downstream effectors in human breast cancer. AB - Phosphatidylinositol 3-kinase (PI3-K) is a growth factor-activated transforming lipid (and protein) kinase, involved in cell motility and invasion, that has multiple effectors. Relatively little is known about its expression and enzymatic activity in human breast cancer. Since growth factor receptors are amplified in breast cancer, and the tumor suppressor PTEN may be mutated in human breast cancer, it was hypothesized that PI3-K and its downstream effectors would be activated in this disease. In 11 resected tumors analyzed for expression of this kinase, a mean 3-fold increase in protein expression was observed over the corresponding adjacent control tissue. Using an in vitro lipid kinase assay of the immunoprecipitated PI3-K protein, a greater than 2-fold increase in activation was observed. These changes were observed in the absence of an activation of either protein kinase B (PKB, akt1) or p70 S6 kinase (p70 S6K). However, p21-activated kinase (Pak), p38 mitogen-activated protein kinase (p38 MAPK) and mitogen-activated protein kinase-activated protein kinase 2 (MAPKAPK 2) were all overexpressed and demonstrated increased enzyme activity. It may be concluded that aberrant mitogenic signaling in human breast cancer in vivo involves Pak, p38 MAPK and MAPKAPK2 downstream of PI3-K, but neither of PKB or p70 S6K. It is proposed that this pathway may serve as a useful targeting nexus for investigation of small molecule inhibitors in human breast cancer. PMID- 11857400 TI - Prevalence of Kaposi's sarcoma-associated herpesvirus infection in sex workers and women from the general population in Spain. AB - Transmission routes of Kaposi's sarcoma-associated herpesvirus (KSHV) in the general population are poorly understood. Whereas sexual transmission appears to be common in homosexual men, the evidence for heterosexual transmission is less convincing. In our study, prevalence of KSHV infection was examined among women in the Spanish general population and among sex workers. Subjects consisted of 100 prostitutes and 100 women randomly sampled from the general population and age-matched to the prostitutes. Women had a personal interview and gynecologic examinations in which a blood sample, cervical cells and oral cells were obtained. Peripheral blood mononuclear cells (PBMC), oral and cervical samples were tested for KSHV DNA by quantitative real-time PCR. Sera were tested for antibodies against human immunodeficiency virus (HIV) by ELISA and against KSHV by latent IFA and K8.1 ELISA. Women who were positive in either serologic assay or PCR were considered infected by KSHV. Human papillomavirus (HPV) DNA in cervical scrapes were evaluated using the Hybrid Capture System. The study population had an average age of 30 years and were HIV-negative. Women from the general population were largely of Spanish nationality, and 61% reported lifetime monogamy. The majority of the prostitutes (76%) were immigrants, primarily from South America. Sex workers were twice as likely to be infected with KSHV than women in the general population (16% vs. 8%, prevalence odds ratio [OR] = 2.2). KSHV was more prevalent among HPV DNA-positive women (OR = 2.5) and among women with an early age at first sexual intercourse (OR = 2.7, p < 0.05). KSHV DNA was detected by PCR in 3% of the oral cavity samples, in 2% of the cervical samples of the prostitutes and in 1% of the cervical samples of women in the general population. All PBMC samples were negative. These results suggest that in low risk countries for KSHV, oral shedding and heterosexual contacts are potential pathways for KSHV transmission. PMID- 11857402 TI - P53: an ubiquitous target of anticancer drugs. AB - The p53 tumor suppressor can induce growth arrest, apoptosis and cell senescence. Not surprisingly, p53 is an appealing target for therapeutic intervention. Although current anticancer agents do not directly interact with p53, these agents (including DNA damaging drugs, antimetabolites, microtubule-active drugs and inhibitors of the proteasome) cause accumulation of wt p53. Depending on the p53 status of cancer cells, diverse therapeutic strategies are under development. These include pharmacological rescue of mutant p53 function and reactivation of wt p53 in E6-expressing cells. For protection of normal cells, strategies range from abrogation of wt p53 induction, thereby decreasing the toxicity of DNA damaging agents, to activation of wt p53-dependent checkpoints, thereby protecting cells against cell cycle-dependent therapeutics. PMID- 11857404 TI - Molecular and functional MDR1-Pgp and MRPs expression in human glioblastoma multiforme cell lines. AB - The aim of our study was to investigate the functional expression of P glycoprotein (Pgp) and multidrug resistance-associated proteins (MRPs) in 2 distinct glioma cells (GL15 and 8MG) from patients with glioblastoma multiforme. MDR1 gene and Pgp expression was not detected in either cell line by RT-PCR and Western blotting, respectively. In contrast, MRP1 was detected at both mRNA and protein level in both cell lines, with a higher expression in the 8MG cells that occur predominantly at the cell membrane. Three other MRPs (MRP3, MRP4 and MRP5) were detected by RT-PCR in both cell lines, whereas MRP2 was not expressed. In addition, MRP3 protein was also detected by immunocytochemistry in both GL15 and 8MG cell lines. Indomethacin and probenecid, 2 modulators of MRPs activity, increased the accumulation of vincristine and etoposide, 2 substrates of MRPs, by both cell lines. These modulators also decreased the efflux of vincristine from both cell lines with a more pronounced effect in 8MG cells. In conclusion, our results show functional expression of MRPs leading to a decrease in the intracellular vincristine and etoposide concentrations in human glioblastoma cell lines. Furthermore, our results that exhibit protein expression of MRP1 and MRP3 and gene expression of MRP4 and MRP5 in these 2 glioblastoma cell lines suggest new mechanisms that could lead to a MDR phenotype of tumour cells in patients with glioblastoma multiforme. PMID- 11857403 TI - Anticarcinogenic effect of FTY720 in human prostate carcinoma DU145 cells: modulation of mitogenic signaling, FAK, cell-cycle entry and apoptosis. AB - Despite the high frequency of prostate cancer, therapeutic options for advanced disease are limited to chemotherapy, radiation or hormonal therapy and eventually fail in all patients. Therefore, alternative approaches need to be developed. We previously reported that FTY720, a metabolite from Isaria sinclarii, is a unique antitumor agent for an androgen-independent prostate cancer cell line and requires caspase-3 activation in apoptosis. In our study, we have evaluated the effect of FTY720 on a family of mitogen-activated protein kinases (MAPKs), focal adhesion kinase (FAK), mitochondrial transmembrane potential, caspase-9 and caspase-8 and analyzed the expression of some cell-cycle regulator proteins in DU145 cells in order to understand the various antitumor effects of FTY720. Apoptosis was quantified by phosphatidylserine exposure. Activation of MAPKs, cleavage of caspase-9 and caspase-8, status of cyclin-dependent kinases (CDKs) and Cip1/p21, a cyclin-dependent kinase inhibitor, were evaluated by Western blot analysis, in addition to FAK and phospho-FAK immunoprecipitation and cell-cycle analysis by FACScan. We found that in DU145 cells, 40 microM FTY720 caused activation of p38 MAPK and the upstream kinase MKK3/MKK6 but not SAPK/JNK. Mitochondrial transmembrane potential, FAK and ERK1/2 were reduced while caspase 9 and caspase-8 were cleaved. The p38-specific inhibitor had no effect on apoptosis induced by FTY720, whereas z-VAD.FMK, a broad-spectrum caspase inhibitor, did not inhibit the p38 MAPK activation. An amount of 20 microM FTY720 resulted in G(1) arrest and a decrease of CDK2 as well as CDK4, whereas it induced Cip1/p21. FTY720 may exert anticarcinogenic effects against prostate cancer cells possibly involving modulation of mitogenic signaling, cell-cycle regulators, induction of G(1) arrest and apoptotic death in DU145 cells. PMID- 11857405 TI - Overexpression of the eukaryotic translation initiation factor 4G (eIF4G-1) in squamous cell lung carcinoma. AB - eIF4G-1 belongs to the family of translational initiation factors and is recognized as the central organizing protein in recruitment of mRNA during translational initiation. Previously published studies have provided some evidence that overexpression of translational factors is a general event in the process of carcinogenesis. We have characterized the expression of the eIF4G-1 protein in 33 squamous cell carcinoma (SCC) of the lung by Western blotting. Overexpression of the eIF4G-1 protein was detected in 61% of the tumors compared to the respective normal lung tissue. In addition, we analyzed the expression of this protein by immunohistochemistry in 138 SCC of the lung using a newly generated antibody that is specific for eIF4G-1 as determined by Western blotting. This anti-eIF4G-1 antibody was suitable for the immunohistochemistry of paraffin-embedded tissues. There is a strong cytoplasmic staining detected in the tumor areas that is consistent with the cytoplasmic localization of the translation factor eIF4G-1. In 72% of the examined tissue sections of SCCs of the lung, we detected an overexpression of the eIF4G-1 protein compared to the surrounding connective tissue. Two tumors that were analyzed by both methods showed an overexpression of eIF4G-1 both with Western blot analysis and immunohistochemical staining. Overexpression of eIF4G-1 may result in an increased amount of the translation initiation complex eIF4F, which in turn may activate the translation of the same target mRNAs as eIF4E. PMID- 11857406 TI - Collagen type I expression in experimental anaplastic thyroid carcinoma: regulation and relevance for tumorigenicity. AB - Fibrosis in solid malignancies plays a significant role in tumor pathophysiology. Potential mechanisms for collagen type I deposition in anaplastic thyroid carcinoma (ATC) were investigated using 6 characterized ATC cell lines. Three of these cell lines, which produced collagen type I, had, as a group, a poor tumorigenicity when inoculated in athymic mice. This group of cells generated tumors in 4 of 24 injected animals (17%). Pro-alpha 1(I) collagen mRNA-expressing carcinoma and stromal cells were interdispersed in the tumors generated by these ATC cells. By contrast, the 3 noncollagen-producing ATC cell lines were all tumorigenic with a tumor take of 60% in the whole group. In the latter tumors, pro-alpha 1(I) collagen mRNA-expressing cells were confined to the stromal compartment, well delineated from carcinoma cell islets. To study the influence of ATC cells on collagen type I synthesis by fibroblasts, we used AG 1518 diploid human fibroblasts cultured on poly-(2-hydroxyethyl methacrylate) (poly[HEMA]) coated plates. This culture condition allows the study of the effect of collagen mRNA translation in the regulation of collagen type I synthesis. Conditioned media from the 6 ATC cell lines did not influence collagen synthesis. The ATC cell line KAT-4 stimulated fibroblast synthesis of collagen type I when the two cell types were cocultured on poly[HEMA]-coated substrates. Specific inhibitors of PDGF and TGF-beta reduced the KAT 4 carcinoma cell-induced stimulation of collagen type I synthesis. Our data suggest that collagen type I production by carcinoma cells correlates negatively with tumorigenicity and that the formation of a well-defined stroma is of importance for tumor growth. Furthermore, our data suggest that tumor cells are able to stimulate collagen mRNA translation in stromal fibroblasts in direct cell-cell contact by, at least in part, transferring PDGF or TGF-beta. PMID- 11857407 TI - Inhibition of constitutive NF-kappa B activity suppresses tumorigenicity of Ewing sarcoma EW7 cells. AB - Ewing sarcoma is 1 of the most aggressive tumors that can affect children and young adults. Despite advances in therapy, the prognosis remains poor emphasizing the need for defining new targets for treatment. We investigated a possible role of nuclear factor-kappa B (NF-kappa B) activity of Ewing sarcoma-derived EW7 cells in their tumorigenicity. In these cells, expression of a degradation resistant form of the inhibitory factor I kappa B alpha inhibited NF-kappa B activity without affecting their in vitro proliferation rate. It causes, however, a remarkable loss of their ability to generate tumors in nude mice that correlates with both a decrease in extracellular matrix (ECM) protein secretion and an acquisition of sensitivity to murine tumor necrosis factor alpha (TNF alpha)-induced apoptosis. These data support the concept that NF-kappa B activity plays a role in the tumorigenicity of Ewing sarcoma cells, identifying NF-kappa B as a potential target for reducing Ewing tumor progression. PMID- 11857408 TI - CDH1 mutations are present in both ductal and lobular breast cancer, but promoter allelic variants show no detectable breast cancer risk. AB - Mutations and diminished expression of the E-cadherin gene (CDH1) have been identified in a number of epithelial malignancies. Although somatic CDH1 mutations were detected in lobular breast cancer with a frequency ranging from 10 56%, CDH1 alterations in more frequent ductal tumors appear to be rare. Here we have analyzed the coding region of CDH1 for mutations using denaturing high performance liquid chromatography and found 4 mutations in 83 ductal carcinomas (5%) and 3 mutations in 25 lobular carcinomas (12%). The germline of 13 patients with familial lobular tumors was also analyzed for mutations, but none were detected. In a case-control study, we also tested whether a variant adenine allele in the promoter polymorphism -161C-->A with a putative influence on the transcriptional activity of CDH1 in vitro confers any detectable risk of breast cancer. No significant difference in the allelic frequency between patients with breast cancer (326/1,152, 28.3%) and controls (190/696, 27.3%, p > 0.05; relative risk 1.05, 95% confidence interval 0.85-1.30) was found. A novel promoter polymorphism was identified at position -152, but the frequency of the variant cytosine allele was also similar in patients with breast cancer and controls (0.71% vs. 0.21%, p = 0.23). Transient transfection experiments using reporter constructs containing the nucleotide substitutions -161C/-152C and -161A/-152T showed only a slight decrease in the transcription activity compared to the wild type construct. These results do not support CDH1 as a prominent low-penetrance cancer susceptibility gene, but indicate that CDH1 mutations contribute to the progression of both lobular and ductal tumors. PMID- 11857409 TI - Sporadic breast cancer in young women: prevalence of loss of heterozygosity at p53, BRCA1 and BRCA2. AB - Previous studies have shown that breast cancers have more aggressive pathologic features in young women. In order to examine genetic alterations associated with early-onset breast cancer, 31 patients with no known family history, aged 26-35 years at diagnosis, were examined for loss of heterozygosity (LOH) at 3 key chromosomal intervals: 17p (p53), 17q 21 (BRCA1) and 13q12-13 (BRCA2) using polymerase chain reaction analysis of polymorphic microsatellite markers. These were compared with 31 patients aged 55-72 years that were matched for size, type and grade. All young breast cancer cases exhibited LOH for at least 1 marker and 20 cases (64.5%) exhibited LOH at 1 or more markers from each interval. The frequency of LOH detected for each of the markers was as follows 17p: p534N (33.3%), D17S796 (36.7%), D17S799 (63.3%) and D17S513 (59.3%); 17q: D17S855 (64.5%), THRA1 (46.7%) and D17S579 (33.3%); and 13q: D13S260 (74.2%), D13S171 (47.6%) and D13S267 (40.0%). These frequencies are higher than those observed at the 3 markers studied in the matched postmenopausal patients: D17S799 (41.4%), D17S855 (35.5%), D13S260 (30.0%). These differences in frequency of LOH were statistically significant for the D17S855 and D13S260 markers (p < 0.025 and p < 0.001 respectively). Although there were more grade III carcinomas (21 of 31 cases), there was no correlation between number of alterations and high grade in younger cases. These data suggest that LOH at these regions could be related to early-onset sporadic breast cancer. PMID- 11857410 TI - Inhibition of metalloproteinases enhances the internalization of anti-CD30 antibody Ki-3 and the cytotoxic activity of Ki-3 immunotoxin. AB - CD30 is selectively expressed on the tumor cells of a variety of malignant disorders of the immune system and can therefore be used as a target for an anti CD30 antibody-based immunotherapy. However, CD30 is cleaved at the cell surface by tumor necrosis factor-alpha converting enzyme (TACE). This metalloproteinase releases the soluble ectodomain of CD30 (sCD30), which is able to neutralize immunotherapeutic agents before these reach their target cells. Such constitutive CD30 cleavage is enhanced after binding of most anti-CD30 antibodies, leading to a downregulation of CD30 and an increase of sCD30 in the cell environment. Here, we demonstrate that CD30 shedding from the cell line Karpas 299 could effectively be blocked by the hydroxamic acid-based metalloproteinase inhibitors BB-3644 (IC50 = 180 nM), BB-2116 (IC50 = 230 nM), BB-94 (batimastat, IC50 = 230 nM) and BB-2516 (marimastat, IC50 = 1 microM). This inhibition reduced the concentration of sCD30 in the cell environment to the background level, prolonged the persistence of the anti-CD30 antibody Ki-3 on Karpas 299 cells and favored its internalization. Moreover, a nontoxic concentration of the inhibitor BB-3644 significantly increased the cytotoxic activity of the anti-CD30 ricin A-chain immunotoxin Ki-3.dgA towards the CD30(+) Hodgkin-derived cell line L540. Hence, the metalloproteinase inhibitor BB-3644 may be a promising compound to improve the immunotherapy of CD30(+) malignancies. PMID- 11857411 TI - Frequencies of HER-2/neu overexpression relating to HLA haplotype in patients with gastric cancer. AB - We have identified that HER-2/neu-derived peptides are naturally processed as tumor rejection antigens recognized by tumor-specific, HLA-A2-restricted cytotoxic T lymphocytes in gastric cancer. To evaluate candidates for immunotherapy using HER-2/neu-derived, HLA-A2-restricted peptides, we examined the frequency of HLA-A2 relating to HER-2/neu overexpression or the infiltrating grade of tumor-infiltrating lymphocytes (TILs) in Japanese patients with gastric cancer. HER-2/neu-overexpressing tumors detected by immunohistochemistry amounted to 19% of primary gastric cancers and HLA-A2-positive patients with gastric cancer were 31% of primary gastric-cancer cases. Finally, gastric-cancer patients with both HLA-A2-positive and HER-2/neu-overexpressing tumors amounted to 6.6% of these cases. There was no significant difference in the infiltrating grade of TILs between gastric cancers overexpressing HER-2/neu and those that did not. The candidate for HER-2/neu-based immunotherapy with HLA-A2-restricted peptides represent a very limited population of Japanese patients. PMID- 11857412 TI - Lipopeptide-based melanoma cancer vaccine induced a strong MART-27-35-cytotoxic T lymphocyte response in a preclinal study. AB - Identification of tumor antigens and their optimal antigenic peptides raised hopes for the development of peptide-based immunotherapeutic vaccine strategies for human melanoma, however. Synthetic peptides alone are not immunogenic enough, and adequate formulation is critical for elaboration of peptide vaccines. To improve formulation, we evaluated 2 lipopeptide constructs, both including HLA-A2 restricted MART 27-35-CD8+ T lymphocyte (CTL) epitope covalently linked to universal tetanus toxoid (TT) 830-843 helper T lymphocyte (HTL) epitope, in HLA A2 transgenic mouse models that mimic human CTL responses in vivo. These 2 constructs only differed in the formulation of their lipid tail. We showed that lipopeptide constructs were strongly recognized, in vitro, by human MART 27-35 cytotoxic T cells derived from tumor-infiltrating lymphocytes. The transgenic Mice immunized with these 2 MART lipopeptide formulations containing covalently linked HTL-CTL epitopes induced strong MART 27-35 cytotoxic T cells. This CTL induction was critically dependent on the presence of the helper T lymphocyte epitope. These results also showed that a single palmitoyl-lysine chain is enough to assure immunogenicity of a given peptide and that the presence of a lipid tail bypass the need for adjuvant. These results support the selection of MART lipopeptide melanoma vaccine for evaluation in a clinical trial. PMID- 11857413 TI - Vascular endothelial growth factor and prognosis in patients with node-negative breast cancer. AB - Vascular endothelial growth factor (VEGF) has emerged as one of the most important angiogenic growth factors from experimental in vitro and in vivo studies. In the present study, we investigated the relationship between VEGF expression and microvessel density (MVD) and defined their prognostic relevance on a series of 242 patients with node-negative breast cancer, using immunohistochemical methods. In parallel, estrogen and progesterone receptors were quantitatively assessed using the dextran-charcoal technique and cell proliferation was evaluated as S-phase cell fraction according to (3)H-thymidine labeling index (TLI). The percentage of VEGF-expressing cells varied from 0-95% in the different tumors and was unrelated to menopausal status, tumor size or steroid receptor status. Conversely, a significant inverse relation was observed with patient age or tumor cell proliferation, albeit with very poor correlation coefficients. A significant relation was observed between VEGF expression and MVD (r(s) = 0.55, p < 0.001). Clinical outcome analyzed as a function of high and low VEGF expression showed slight differences in terms of both disease-free survival (DFS) and overall survival (OS) that never reached statistical significance. Moreover, the trend was paradoxically in favor of patients with highly VEGF expressing tumors. Finally, DFS and OS curves, when analyzed as a function of VEGF expression or MVD, were superimposable. In conclusion, our study did not highlight a prognostic relevance of VEGF expression in patients with node negative breast cancer, as already observed for MVD. PMID- 11857414 TI - Curcumin exerts multiple suppressive effects on human breast carcinoma cells. AB - In our study, we present experimental evidence suggesting that curcumin exerts multiple different suppressive effects on human breast carcinoma cells in vitro. Our experiments demonstrate that curcumin's antiproliferative effects are estrogen dependent in ER (estrogen receptor)-positive MCF-7 cells, being more pronounced in estrogen-containing media and in the presence of exogenous 17-beta estradiol. Curcumin inhibits the expression of ER downstream genes including pS2 and TGF-beta (transforming growth factor) in ER-positive MCF-7 cells, and this inhibition is also dependent on the presence of estrogen. Curcumin also decreases ERE (estrogen responsive element)-CAT activities induced by 17-beta estradiol. In addition, we demonstrate that curcumin exerts strong anti-invasive effects in vitro that are not estrogen dependent in the ER-negative MDA-MB-231 breast cancer cells. These anti-invasive effects appear to be mediated through the downregulation of MMP-2 (matrix metalloproteinase) and the upregulation of TIMP-1 (tissue inhibitor of metalloproteinase), 2 common effector molecules that have been implicated in regulating tumor cell invasion. Our study also demonstrates that curcumin inhibits the transcript levels of 2 major angiogenesis factors VEGF (vascular endothelial growth factor) and b-FGF (basic fibroblast growth factor) mainly in ER-negative MDA-MB-231 cells. PMID- 11857415 TI - Meat consumption and colorectal cancer risk: dose-response meta-analysis of epidemiological studies. AB - The hypothesis that consumption of red and processed meat increases colorectal cancer risk is reassessed in a meta-analysis of articles published during 1973 99. The mean relative risk (RR) for the highest quantile of intake vs. the lowest was calculated and the RR per gram of intake was computed through log-linear models. Attributable fractions and preventable fractions for hypothetical reductions in red meat consumption in different geographical areas were derived using the RR log-linear estimates and prevalence of red meat consumption from FAO data and national dietary surveys. High intake of red meat, and particularly of processed meat, was associated with a moderate but significant increase in colorectal cancer risk. Average RRs and 95% confidence intervals (CI) for the highest quantile of consumption of red meat were 1.35 (CI: 1.21-1.51) and of processed meat, 1.31 (CI: 1.13-1.51). The RRs estimated by log-linear dose response analysis were 1.24 (CI: 1.08-1.41) for an increase of 120 g/day of red meat and 1.36 (CI: 1.15-1.61) for 30 g/day of processed meat. Total meat consumption was not significantly associated with colorectal cancer risk. The risk fraction attributable to current levels of red meat intake was in the range of 10-25% in regions where red meat intake is high. If average red meat intake is reduced to 70 g/week in these regions, colorectal cancer risk would hypothetically decrease by 7-24%. PMID- 11857416 TI - Ultrasound screening and risk factors for death from hepatocellular carcinoma in a high risk group in Taiwan. AB - Although previous studies have demonstrated the ability of ultrasonography (US) screening to detect small asymptomatic hepatocellular carcinoma (HCC), the efficacy of US screening in reducing deaths from HCC still remained unresolved. A 2-stage screening program was designed to identify a high risk group in 7 townships in Taiwan by 6 markers (of risk for HCC) and repeated US screening was further applied to those with at least 1 positive result for the 6 markers, with a range of 3- to 6-month inter-screening intervals to those with liver cirrhosis or other chronic liver diseases and an annual screening regime for the remaining subjects with normal findings according to US. The 4,843 subjects in this cohort were followed up for an average of 7 years. We compared 4,385 attenders with 458 non-attenders, in conjunction with baseline assessment for self-selection bias. In addition, we assessed baseline variables with respect to their effects on risk of incidence of and mortality from HCC and on risk of incidence of liver cirrhosis. The difference in mortality between attenders and non-attenders was then re-estimated adjusting for significant predictors of cirrhosis, HCC incidence and HCC death as a further guard against baseline differences between attenders and non-attenders in risk profiles. Results of US screening for this high risk group found the mortality was lower by 24% (95% CI: -52 to 62%) in the attenders compared to the non-attenders. After adjustment for sensitivity, the mean sojourn time (MST) were 1.57 (95% CI: 0.94-4.68) for subjects with liver cirrhosis and 2.66 (95% CI: 1.68-6.37) years for non-cirrhotic patient. Significant increases in risk of HCC incidence were associated with increasing age, male gender, hepatitis B surface antigen positive (HbsAg), hepatitis C antibody positive (Anti-HCV), high levels of alanine transaminase (ALT) and alpha fetoprotein (AFP) and a family history of HCC. Significantly increased risks of liver cirrhosis were associated with predictors of cirrhosis were increasing age, HbsAg, high levels of ALT and of AFP. Significant or borderline significant increases in risk of HCC death were associated with increasing age, male gender, HbsAg, high levels of AST and AFP. Adjusted for the significant variables, the mortality was lower by 41% (95% CI: -20 to 71%, p = 0.1446) in the attenders compared to the non-attenders. The present study provides suggestive evidence on the efficacy of US screening in a selective high risk group in an endemic area of hepatitis B. A randomized controlled trial would yield definitive evidence. Within the protocol of such a trial, a shorter interscreening interval for patients with liver cirrhosis is suggested. PMID- 11857417 TI - Incidence and mortality of testicular and prostatic cancers in relation to world dietary practices. AB - The incidence and mortality rates of testicular and prostatic cancers in 42 countries were correlated with the dietary practices in these countries using the cancer rates (1988-92) provided by the International Agency for Research on Cancer (IARC) and the food supply data (1961-90) provided by the Food and Agriculture Organization (FAO). Among the food items we examined, cheese was most closely correlated with the incidence of testicular cancer at ages 20-39, followed by animal fats and milk. The correlation coefficient (r) was highest (r = 0.804) when calculated for cheese consumed during the period 1961-65 (maternal or prepubertal consumption). Stepwise-multiple-regression analysis revealed that milk + cheese (1961-65) made a significant contribution to the incidence of testicular cancer (standardized regression coefficient [R] = 0.654). Concerning prostatic cancer, milk (1961-90) was most closely correlated (r = 0.711) with its incidence, followed by meat and coffee. Stepwise-multiple-regression analysis identified milk + cheese as a factor contributing to the incidence of prostatic cancer (R = 0.525). The food that was most closely correlated with the mortality rate of prostatic cancer was milk (r = 0.766), followed by coffee, cheese and animal fats. Stepwise-multiple-regression analysis revealed that milk + cheese was a factor contributing to mortality from prostatic cancer (R = 0.580). The results of our study suggest a role of milk and dairy products in the development and growth of testicular and prostatic cancers. The close correlation between cheese and testicular cancer and between milk and prostatic cancer suggests that further mechanistic studies should be undertaken concerning the development of male genital organ cancers. PMID- 11857418 TI - Prostate cancer mortality reduction by screening: power and time frame with complete enrollment in the European Randomised Screening for Prostate Cancer (ERSPC) trial. AB - From 1992-2001, 7 countries in Europe gradually recruited men for the European Randomised Screening for Prostate Cancer (ERSPC) trial. Centres recruit different age groups and have different designs for recruiting and countries have different underlying risks for prostate cancer. Recruitment has reached 163,126 men aged 55 69 at entry now. Our purpose was to calculate the power of the trial and at what point in time can statistically significant differences in prostate cancer mortality be expected. Recruitment data were collected from the screening centres. We calculated the expected number of prostate cancer deaths in each follow-up year, based on national statistics and expected rate in trial entrants. The power was calculated using different assumptions on intervention effect and contamination rate and also if the ERSPC trial would cooperate with other trials. With an assumed 25% intervention effect in men actually screened and a 20% contamination rate, the trial will reach a power of 0.86 in 2008. With an assumed intervention effect of 40%, the power reaches 0.90 in 2003-2004. Pooling data with those of the Prostate, Lung, Colorectal and Ovary (PLCO) trial early is expected to improve the power to 79% (20% intervention effect) to 92% (40% intervention effect PLCO). Adding more centres with compliance rates lower than 45% decreases the power of the trial. The ERSPC trial has sufficient power to detect a significant difference in prostate cancer mortality between the 2 arms if the true reduction in mortality by screening is 25% or more or if contamination remains limited to 10% if the true effect is 20% or more. If early detection and treatment turns out to have a stronger effect as may be suggested by observational data, the ERSPC trial is likely to conclusively show that within the next 5 years. PMID- 11857419 TI - Familial aggregation of urothelial cell carcinoma. AB - Urothelial cell carcinoma (UCC) is not considered to be a familial disease. Familial clustering of UCC was described in several case reports, however, some with an extremely early age at onset suggesting a genetic component. Epidemiological studies yielded inconsistent evidence of familial UCC, possibly because of low power and the inability to adjust for strong confounding. In our study the existence of a familial subtype of UCC was evaluated, as well as familial clustering of UCC with other types of cancer. A population-based family case-control study was performed including patients newly diagnosed with UCC of the bladder, ureter, renal pelvis or urethra, between January 1995 and December 1997, in the southeastern part of the Netherlands. Information on the patients' first-degree relatives was collected by postal questionnaire and subsequent telephone calls. The patients' partners filled out a similar questionnaire on their relatives. All reported occurrences of UCC were verified using medical records. Disease occurrence among case-relatives and control-relatives was compared to obtain the familial risk. Random effect proportional hazards regression analyses were used to calculate this familial risk while adjusting for age, gender and smoking behavior. In 95 families of the 1,193 patients and in 36 families of the 853 partners at least 1 relative was diagnosed with UCC. This yielded an adjusted hazard ratio (HR) of 1.8 (95% CI: 1.3-2.7). An increased risk was also found for cancer of the hematolymphopoietic system (hazard ration = 1.9, 95% CI: 1.2-3.1) among case-relatives. These results indicate that UCC has a familial component with an almost 2-fold increased risk among first-degree relatives of patients with UCC, which cannot be explained by smoking. Future segregation analyses may indicate whether this clustering can be attributed to genetic susceptibility. PMID- 11857420 TI - Body mass and stage of breast cancer at diagnosis. AB - Obesity is a well-known risk factor for postmenopausal breast cancer. In contrast, the relationship between obesity and stage of breast cancer at diagnosis is less clear. We hypothesized that increased breast size in obese women may delay discovery of breast tumors. Thus, the purpose of our study was to examine whether there is an association between body mass and stage of breast cancer at diagnosis using hospital medical records. Newly diagnosed breast cancer cases (n = 966) in the Baltimore metropolitan area from 1991 to 1997 were included in our study. Patient information including age, ethnicity, weight, height and pathology data were obtained from hospital medical records. High body mass was significantly associated with late stage of breast cancer at diagnosis. Women who were obese (body mass index [BMI] > or = 27.3) were more likely to be at an advanced stage at diagnosis compared with women with a BMI of < 27.3 (multivariate-adjusted odds ratio [OR] 1.57, 95% confidence interval [CI] 1.15 2.14). The association between body mass and stage at diagnosis was stronger among women younger than 50 years (OR 2.34, 95% CI 1.34-4.08) compared with women 50 years or older (OR 1.30, 95% CI 0.89-1.91). Our study suggests that higher body mass is associated with advanced stage of breast cancer at diagnosis. This finding may be of considerable concern, given the increasing prevalence of obesity in women in the United States and the poor prognosis associated with late stage tumors. PMID- 11857421 TI - Photodynamic therapy with hypericin induces vascular damage and apoptosis in the RIF-1 mouse tumor model. AB - Hypericin, a polycyclic quinone obtained from plants of the genus Hypericum, has been proven to be a potent photosensitizer. The mechanism of tumor eradication and mode of cell death induced by in vivo photodynamic therapy (PDT) with hypericin were investigated in the present study using 2 therapeutic protocols. RIF-1 tumors were exposed to laser light at either 0.5 hr or 6 hr after hypericin administration (5 mg/kg, i.v.). A significant reduction in tumor perfusion, as determined by the retention of fluorescein in the tumor tissue, was detected immediately after both PDT treatments. Further decrease in tumor perfusion was observed in the hours after treatment. The re-establishment of tumor perfusion, however, occurred 24 hr after 6 hr-interval PDT, but not after 0.5 hr-interval PDT. The kinetics of tumor cell survival estimated by the in vivo/in vitro clonogenic assay revealed no or limited cell death when tumors were explanted immediately after irradiation, whereas a delayed but progressive cell death was detected when tumors remained in situ after both PDT treatments. The detection of nucleosomal DNA fragmentation by agarose gel electrophoresis or TUNEL assay and the assessment of cell morphology by light microscopy indicated that apoptosis was the most prominent tumor response to hypericin-mediated PDT. Furthermore, immunohistochemical analysis of the tumor tissue showed an increased expression of both Fas and Fas ligand after irradiation, suggesting that this cell death pathway might contribute to the overall PDT-induced apoptotic response. In conclusion, our results demonstrate that apoptosis, likely occurring as a result of vascular damage, is responsible for the tumor eradication by PDT with hypericin in this tumor model. PMID- 11857422 TI - Use of the human MDR1 promoter for heat-inducible expression of therapeutic genes. AB - The promoter of the human multidrug resistance gene (mdr1) harbors stress responsive elements, which can be induced e.g., by heat or cytostatic drugs. In previous studies the drug-responsiveness of the mdr1 promoter was successfully used for the drug-inducible expression of the human TNF-alpha gene in vitro and in vivo. Beside the drug-responsive elements of the mdr1 promoter, heat-shock responsive elements have also been identified, which could be exploited for construction of heat-inducible expression vectors. To analyze the hyperthermia inducibility of the mdr1 promoter we used the pmdr-p-CAT and pM3mdr-p-hTNF vector constructs. Both constructs carry the mdr1 promoter fragment spanning from -207 to +153 to drive expression of the CAT-reporter or TNF-alpha gene. We tested the heat-induced CAT-reporter and TNF-alpha expression in vitro in transduced HCT15 and HCT116 human colon carcinoma cells. For the studies the transduced tumor cells were treated with hyperthermia at 41.5 degrees C or 43 degrees C for 2 hr to induce CAT or TNF-alpha expression. Cells and supernatants were harvested before hyperthermia and at certain time points (0-120 hr) after heat shock. The heat-induced CAT-reporter expression or TNF-alpha secretion was determined by specific ELISA. The experiments indicate that hyperthermia activates the mdr1 promoter in a temperature and time dependent manner. This induction leads to an 2 to 4-fold increase in CAT-reporter or 2- to 7-fold increase in TNF alpha expression in the tumor cell lines. These experiments reveal that the mdr1 promoter driven expression of therapeutic genes can be employed for combined cancer gene therapy approaches. PMID- 11857423 TI - Inhibition of cell proliferation, invasion, tumor growth and metastasis by an oral non-antimicrobial tetracycline analog (COL-3) in a metastatic prostate cancer model. AB - Antibiotic forms of tetracycline exhibit antitumor activity in some tumor models. However, their low in vivo efficacy and associated morbidity limit their long term application in cancer therapy. This report appraises the efficacy of doxycycline (DC) and non-antimicrobial, chemically modified tetracyclines (CMTs) against prostate cancer. Both DC and several CMTs inhibited prostate tumor cell proliferation in vitro. Some of the CMTs were significantly more potent than DC. One of the CMTs, 6-deoxy, 6-demethyl, 4-de-dimethylamino tetracycline (CMT-3, COL 3), was the most potent inhibitor (50% inhibition dose [GI(50)] < or = 5.0 ,microg/ml). Exposure of tumor cells to CMT-3 induced both apoptosis and necrosis. Mitochondrial depolarization and increased levels of reactive hydroxyl radicals were also observed in cells treated with CMT-3. Cell cycle arrest at the G(0)/G(1) compartment was observed in CMT-3- and DC-treated cells. DC and CMTs also inhibited the invasive potential of the tumor cells in vitro, from 10% (CMT 6) to >90% (CMT-3). CMT-3 and DC decreased matrix metalloproteinase (MMP)-2, tissue inhibitor of MMP (TIMP)-1 and TIMP-2 secretion in treated cultures and inhibited activity of secreted MMPs, CMT-3 was a stronger inhibitor. Daily oral gavage of DC and CMT-3 inhibited tumor growth and metastasis in the Dunning MAT LyLu rat prostate tumor. Decreases in tumor growth (27-35%) and lung metastases were observed (28.9 +/- 15.4 sites/animal [CMT-3-treated] versus 43.6 +/- 18.8 sites/animal [DC-treated] versus 59.5 +/- 13.9 [control]; p < 0.01]. A delay in tumor growth (27 +/- 9.3%, p < 0.05), reduction in metastases (58 +/- 8%) and decrease in tumor incidences (55 +/- 9%, CMT-3-treated) were also observed, when rats were predosed for 7 days. No significant drug-induced morbidity was observed in any of the animals. These results, along with a recently concluded clinical trial, suggest a potential use of CMT-3 as an oral, nontoxic drug to treat metastatic prostate and other cancers. PMID- 11857424 TI - Antitumor activity of the selective epidermal growth factor receptor-tyrosine kinase inhibitor (EGFR-TKI) Iressa (ZD1839) in an EGFR-expressing multidrug resistant cell line in vitro and in vivo. AB - Selective tyrosine kinase inhibitors are regarded as promising antitumor agents for cancer treatment. Iressa (ZD1839) is an orally active, selective EGFR-TKI (epidermal growth factor receptor-tyrosine kinase inhibitor) that blocks signal transduction pathways implicated in cancer cell proliferation, survival and other host-dependent processes promoting cancer growth. The cellular mechanisms of ZD1839 action against human malignant cells and drug-resistant cells were evaluated in vitro. Among the cell lines tested, ZD1839 showed a strong growth inhibitory effect in vitro on human leukemic cells resistant to phorbol ester. This cell line, K562/TPA, shows a non-P-glycoprotein-mediated multidrug-resistant phenotype. The IC50 value of ZD1839 on K562/TPA was approximately 400-fold lower than that on the parental K562 cell (K562 = 12 +/- 2 microM; K562/TPA = 0.025 +/- 0.002 microM) in vitro as determined by a dye formation assay. The expression of EGFR and EGFR mRNA was clearly present in K562/TPA but not in parental K562 cells as determined by Western blotting and RT-PCR. EGFR was autophosphorylated in K562/TPA detected by the antiphosphotyrosine antibody. The in vivo antitumor effects of ZD1839 on K562 and K562/TPA cells were also investigated in BALB/c nude mice. K562/TPA cells transplanted subcutaneously into mice disappeared completely with ZD1839 treatment (20 mg/kg/day, days 3-9). This was not the case in K562 cells. These results suggest that ZD1839 is highly active against tumor cells with non-P-glycoprotein-mediated multidrug resistance that express EGFR. Iressa is a trademark of AstraZeneca (Cheshire, UK). PMID- 11857425 TI - Life style and cancer: effect of divorce. PMID- 11857427 TI - Alginate type and RGD density control myoblast phenotype. AB - Alginates are being increasingly used for cell encapsulation and tissue engineering applications; however, these materials cannot specifically interact with mammalian cells. We have covalently modified alginates of varying monomeric ratio with RGD-containing cell adhesion ligands using carbodiimide chemistry to initiate cell adhesion to these polymers. We hypothesized that we could control the function of cells adherent to RGD-modified alginate hydrogels by varying alginate polymer type and cell adhesion ligand density, and we have addressed this possibility by studying the proliferation and differentiation of C2C12 skeletal myoblasts adherent to these materials. RGD density on alginates of varying monomeric ratio could be controlled over several orders of magnitude, creating a range of surface densities from 1-100 fmol/cm(2). Myoblast adhesion to these materials was specific to the RGD ligand, because adhesion could be competed away with soluble RGD in a dose-dependent manner. Myoblast proliferation and differentiation could be regulated by varying the alginate monomeric ratio and the density of RGD ligands at the substrate surface, and specific combinations of alginate type and RGD density were required to obtain efficient myoblast differentiation on these materials. PMID- 11857428 TI - Resorbability and solubility of zinc-containing tricalcium phosphate. AB - Using zinc-containing tricalcium phosphate (ZnTCP) as the zinc carrier for zinc releasing calcium phosphate ceramic implants promoted bone formation around the implants. Because no quantitative information was available on the equilibrium solubility and resorbability of ZnTCP, in vitro equilibrium solubility and in vivo resorbability of ZnTCP were determined and compared quantitatively in this study. The solubility of ZnTCP decreased with increasing zinc content. The negative logarithm of the solubility product (K(sp)) of ZnTCP was expressed as pK(sp) = 28.686 + 1.7414C - 0.42239C(2) + 0.063911C(3) - 0.0051037C(4) + 0.0001595C(5) in air, where C is the zinc content in ZnTCP (mol %). The solubility of ZnTCP containing a nontoxic level of zinc (<0.63 wt %) decreased to 52-92% of the solubility of pure tricalcium phosphate (TCP) in the pH range 5.0 7.4. However, the in vivo resorbed volume of ZnTCP containing the same amount of zinc was much lower than that expected from the in vitro solubility, becoming as low as 26-20% of that of TCP. Cellular resorption of TCP is substantially a process of dissolution in a fluid with an acidic pH that is maintained by the activities of cells. Therefore, the reduction of the resorbability of ZnTCP could be attributable principally to its lowered cellular activation property relative to that associated with pure TCP. PMID- 11857429 TI - Fabrication of porous hydroxyapatite bodies by a new direct consolidation method: starch consolidation. AB - A new direct consolidation method known as "starch consolidation" was developed especially for the fabrication of porous ceramics, as adopted to prepare hydroxyapatite (OHAp) bodies with different pore sizes and pore volume fractions. The method is based on the swelling ability of starch when it is heated to 80 degrees C in the presence of water. An OHAp powder prepared by a precipitation method and heat treated at 900 degrees C to obtain an appropriate specific surface area for colloidal processing was used in the present work. Wet ball milling was required to deagglomerate the powder and improve its processing ability. Different starch volume fractions and starch particle sizes were added to the OHAp slurries, which were then heat treated to promote consolidation by starch gelification. A linear relationship between the planned and measured porosity values was observed. SEM pictures reveal the presence of interconnected pores with shape and sizes corresponding to starch granules used. Flexural strength varied from about 2 to 15 MPa, for pore volume fractions of approximately 70 and 45%, respectively. A degradation study in an acellular synthetic body fluid (SBF) has shown that the prepared bodies keep their integrity under physiological conditions during the studied time. The method offers possibilities of manufacturing materials that can be used as scaffolds engineering and/or systems for controlled delivery of drugs. PMID- 11857431 TI - Chemistry and biocompatibility of alginate-PLL capsules for immunoprotection of mammalian cells. AB - Transplantation of encapsulated living cells is a promising approach for the treatment of a wide variety of diseases. Large-scale application of the technique, however, is hampered by insufficient biocompatibility of the capsules. In order to get means to study factors influencing the biocompatibility of capsule for encapsulation of living cells, we have correlated the chemical composition of the surface of commonly applied alginate-PLL capsules with the biological response in rats. Capsules prepared of alginates with an intermediate guluronic (G) acid content proved to be biocompatible, whereas capsules prepared of high-G alginates were overgrown by inflammatory cells. We applied X-ray photoelectron spectroscopy to correlate the biological responses with the chemical compositions of the capsule surfaces. High-G alginate capsules proved to have a higher PLL content but less surface binding sites for PLL than low-G alginates. This study, shows for the first time that biological responses against capsules can be successfully correlated to its chemical characteristics. PMID- 11857430 TI - Bone formation in transforming growth factor beta-1-coated porous poly(propylene fumarate) scaffolds. AB - This study determined the bone growth into pretreated poly(propylene fumarate) (PPF) scaffolds implanted into a subcritical size, rabbit cranial defect. PPF scaffolds were constructed by using a photocrosslinking-porogen leaching technique. These scaffolds were then either prewetted (PPF-Pw), treated with RF glow-discharge (PPF-Gd), coated with fibronectin (PPF-Fn), or coated with rhTGF beta1 (PPF-TGF-beta1). One of each scaffold type was then placed into the cranium of nine rabbits. The rabbits were sacrificed after 8 weeks, and the scaffolds were retrieved for histological analysis. The most bone formation was present in the PPF-TGF-beta1 implants; the newly formed bone had a trabecular appearance together with bone marrow-like tissue. Little or no bone formation was observed in implants without rhTGF-beta1. These histological findings were confirmed by image analysis. Bone surface area, bone area percentage, pore fill percentage, and pore area percentage were significantly higher in the rhTGF-beta1-coated implants than in the noncoated implants. No statistical difference was seen between the PPF-Fn, PPF-Pw, or PPF-Gd scaffolds for these parameters. Quadruple fluorochrome labeling showed that in PPF-TGF-beta1 implants bone formation mainly started in the interior of a pore and proceeded toward the scaffold. We conclude that (a) PPF-TGF-beta1 scaffolds can indeed adequately induce bone formation in porous PPF, and (b) PPF scaffolds prepared by the photocrosslinking-porogen leaching technique are good candidates for the creation of bone graft substitutes. PMID- 11857432 TI - In vitro and in vivo dissolution behavior of a dysprosium lithium borate glass designed for the radiation synovectomy treatment of rheumatoid arthritis. AB - Dysprosium lithium borate (DyLB) glass microspheres were investigated for use in the radiation synovectomy treatment of rheumatoid arthritis. In vitro testing focused on weight loss and cation dissolution from glass microspheres immersed in simulated synovial fluid (SSF) at 37 degrees C for up to 64 days. In vivo testing was performed by injecting glass microspheres into the stifle joints of Sprague Dawley rats and monitoring the biodegradability of the microspheres and the tissue response within the joints. The DyLB microspheres reacted nonuniformly in SSF with the majority of lithium and boron being dissolved, whereas nearly all of the dysprosium (>99.7%) remained in the reacted microspheres. Because the DyLB glasses released negligible amounts of dysprosium while reacting with SSF, they are considered safe for radiation synovectomy from the standpoint of unwanted radiation release from the joint capsule. Furthermore, the DyLB microspheres fragmented, degraded, and reacted with body fluids while in the joints of rats without histologic evidence of joint damage. PMID- 11857433 TI - Effects of enamel matrix derivative to titanium implantation in rat femurs. AB - The effects of enamel matrix derivative (EMD; Emdogain) on new trabecular bone induction after pure bioinert titanium (Ti) implantation in the rat femur were examined by means of routine light and transmission electron microscopy, immunohistochemistry, and backscattered electron image analysis. Newly designed mini-Ti implants (3.5 mm in length and 1.6 mm in diameter) were placed in the corticotrabecular area of the femur with either EMD or its carrier, propylene glycol alginate, as control. On post-implantation days 4, 7, 14, and 30, the dissected femur was examined in the transverse direction through Ti implants. In both control and EMD-applied femurs, trabecular bone formation was recognized over the implant surfaces and within medullary cavities even at 4 days post implantation. These newly formed bone trabeculae around the Ti implants were immunoreactive for bone sialoproteins as a bone matrix marker, and osteoclastic bone resorption became evident in these bone trabeculae after 7 days post implantation. Although trabecular bone area around the implants was markedly decreased at 30 days post-implantation compared with those at 14 days, the trabecular bone areas in EMD-applied femurs were significantly greater than those in propylene glycol alginate-applied femurs at both 14 and 30 days post implantation. Our results suggest that EMD is an effective biological matrix for enhancing new trabecular bone induction and resulting attachment of orthopedic prostheses to the recipient bone. PMID- 11857434 TI - Apatite-forming ability of a zirconia/alumina nano-composite induced by chemical treatment. AB - Induction of an apatite-forming ability on a nano-composite of a ceria-stabilized tetragonal zirconia polycrystals (Ce-TZP) and alumina (Al2O3) polycrystals via chemical treatment with aqueous solutions of H3PO4, H2SO4, HCl, or NaOH has been investigated. The Ce-TZP/Al2O3 composite is attractive as a load-bearing bone substitute because of its mechanical properties. The chemical treatments produced Zr-OH surface functional groups, which are known to be effective for apatite nucleation in a body environment. The composite, after chemical treatment, was shown to form a bonelike apatite layer when immersed in a simulated body fluid containing ion concentrations nearly equal to those in human blood plasma. This implies that it may form apatite in the living body and bond to living bone through the apatite layer. This type of bioactive Ce-TZP/Al2O3 composite is therefore expected to be useful as a bone substitute, even under load-bearing conditions. PMID- 11857435 TI - Hemocompatibility of materials used in microelectromechanical systems: platelet adhesion and morphology in vitro. AB - Microelectromechanical systems (MEMS) create an opportunity for the development of smaller, cheaper, and more precise biomedical instrumentation and devices. Little is known, however, about the hemocompatibility of the materials used to fabricate these devices. Because of the potentially harmful consequences of thrombus formation, a better understanding of blood interactions with bioMEMS materials is desirable. This study is an in vitro assessment of the hemocompatibility of silicon (Si), silicon dioxide (SiO2), silicon nitride (Si3N4), low-stress silicon nitride (Si(1.0)N(1.1)), SU-8 photoresist, and parylene thin films. A polycarbonate-based polyurethane, was used as a reference material. Experiments were carried out to detect differences in platelet adhesion or morphology after contact with these materials under static conditions. Platelet adhesion on Si, Si3N4, Si(1.0)N(1.1,) and SU-8 photoresist was significantly greater (P < 0.05) than platelet adhesion on polyurethane. Adhesion on parylene and SiO(2) was not significantly different from on polyurethane (P < 0.05). The median platelet area and circularity were higher on polyurethane than all other materials. Materials that showed higher levels of platelet adhesion tended to have platelets that showed less spreading, except for SiO2, where platelets exhibited relatively low adhesion and spreading. This data suggests that Si, Si3N4, Si(1.0)N(1.1), and SU-8 photoresist may be more reactive to platelets and therefore more thrombogenic than parylene, SiO2, and polyurethane. These results may be helpful in guiding the selection of materials for use in the development of blood-contacting microelectromechanical systems. PMID- 11857436 TI - Effects of pH on human bone marrow stromal cells in vitro: implications for tissue engineering of bone. AB - The objective of this study was to address the hypothesis that changes in extracellular pH alter collagen gene expression, collagen synthesis, and alkaline phosphatase activity in bone marrow stromal cells (BMSCs). Potential effects of pH on cell function are of particular importance for tissue engineering because considerable effort is being placed on engineering biodegradable polymers that may generate a local acidic microenvironment on degradation. Human and murine single-cell marrow suspensions were plated at a density of 2 x 10(4) cells/cm(2). After 7 days in culture, the pH of the culture medium was adjusted to one of six ranges: > or = 7.8, 7.5.-7.7, 7.2-7.4, 6.9-7.1, 6.6-6.8, or < or = 6.5. After 48 h of exposure to an altered pH, alkaline phosphatase activity and collagen synthesis decreased significantly with decreasing pH. This decrease was two-to threefold as pH decreased from 7.5 to 6.6. In contrast, alpha1(I) procollagen mRNA levels increased two- to threefold as pH was decreased. The trend in osteocalcin mRNA expression was opposite to that of collagen. Small shifts in extracellular pH led to significant changes in the ability of BMSCs to express markers of the osteoblast phenotype. These pH effects potentially relate to the microenvironment supplied by a tissue-engineering scaffold and suggest that degrading polymer scaffolds may influence the biologic activity of the cells in the immediate environment. PMID- 11857437 TI - Analysis of acid-treated dentin smear debris and smear layers using confocal Raman microspectroscopy. AB - Smear layers are generally present on any dentin surface prepared with cutting instruments and are often the only available substrate for bonding. It is commonly reported that acid removes these layers, but to date there has been no chemical evidence to support this observation. Confocal Raman microspectroscopy was used to investigate changes in the composition and molecular structure of acid-treated smear debris and in situ dentin smear layers. The exposed dentin in human molars was abraded with 600-grit silicon carbide sandpaper. Raman spectra were acquired on the smear debris and collected from the sandpaper before and after treatment with 10% citric acid, 35% H3PO4, or 0.5M ethylenediaminetetraacetic acid (EDTA). The resultant smeared dentin samples were treated with one of the aforementioned reagents, and spectra were acquired at 1.0 microm intervals across the interfaces of the smear layers/demineralized dentin/mineralized dentin. Corresponding specimens were morphologically analyzed with scanning electron microscopy (SEM). The results showed that the composition of the smear debris and the in situ smear layers was a mixture of disorganized collagen and mineral. Spectral changes in the smear debris suggested that the disorganized collagen was denatured by acid treatment. The denatured collagen formed a gelatinous matrix around the mineral in the smear layer, thereby shielding it from the acid. The smear layers were not apparent in the SEM micrographs of acid-etched dentin prepared and processed with conventional techniques. The micro-Raman spectroscopic results presented in this study provide the first direct evidence that partially denatured collagen within smear layers is not removed and that the mineral is only partially removed with acids that represent conventional dentin adhesive etchants. PMID- 11857438 TI - Adhesion between poly(ethylene-co-vinyl alcohol) (EVA) and titanium. AB - Lap shear adhesive strength between titanium and various kinds of commercial polymers was evaluated. Among them, poly(ethylene-co-vinyl alcohol) (EVA) showed the highest strength. The results of electron spectroscopy for chemical analysis and the contact angle to water indicated that the high adhesive strength of EVA might be due to its high hydrophilicity. Water resistance of adhesion by water immersing at 37 degrees C was investigated. In the case of polyurethane-titanium, the adhesive strength decreased immediately. In contrast, EVA-titanium kept its initial adhesive strength for at least up to 1 month. It was confirmed that surface modification of titanium by hydrogen peroxide enhanced the adhesive and peeling strength. It was based on not only an increase in surface adhesive area but also an increase in the hydrophilicity of titanium by the production of Ti OH. PMID- 11857439 TI - Effect of sodium hypochlorite on dentin bonding with a polyalkenoic acid containing adhesive system. AB - The purpose of this study was to evaluate the effect of sodium hypochlorite (NaOCl) treatment on dentin bonding by means of contact angle (CA), shear bond strength (SBS), and microleakage (ML) measurements. Ultrastructure and nanoleakage (NL) of the interfaces were examined by transmission electron microscopy (TEM). For CA, SBS, and TEM evaluation, human molars were sectioned to expose dentin surfaces and were either acid-etched (35% H3PO4) or further treated with 5% NaOCl for 2 min before the application of Single Bond adhesive. CAs were measured using the Axisymmetric Drop Shape Analysis technique. The Watanabe testing assembly was used for SBS evaluation. ML was assessed by a dye penetration method. NL was examined using a silver-staining technique. The results showed that CA values decreased after acid etching and even more after NaOCl treatment. NaOCl treatment produced lower SBS than acid-etched dentin. Both ML values and NL manifestations were similar for NaOCl-treated and acid-etched dentin. NaOCl did not completely remove the collagen matrix. NL was manifested along the base of hybrid layers and within the polyalkenoic acid copolymer in both groups. Adverse chemical interactions could have occurred between the remnant collagen matrix and/or mineralized dentin after NaOCl treatment. There is no additional advantage in using NaOCl treatment with this adhesive. PMID- 11857440 TI - Biomechanical comparison of the sandblasted and acid-etched and the machined and acid-etched titanium surface for dental implants. AB - To make a direct biomechanical comparison between the sandblasted and acid-etched surface (SLA) and the machined and acid-etched surface (MA), a well-established animal model for implant removal torque testing was employed, using a split-mouth experimental design. All implants had an identical cylindrical solid-screw shape with the standard ITI thread configuration, without any macroscopic retentive structures. After 4, 8, and 12 weeks of bone healing, removal torque testing was performed to evaluate the interfacial shear strength of each surface type. Results showed that the SLA surface was more powerful in enhancing the interfacial shear strength of implants in comparison with the MA surface. Removal torque values of the SLA-surfaced implants were about 30% higher than those of the MA-surfaced implants (p = 0.002) except at 4 weeks, when the difference was at the threshold of statistical significance (p = 0.0519). The mean removal torque values for the SLA implants were 1.5074 Nm at 4 weeks, 1.8022 Nm at 8 weeks, and 1.7130 Nm at 12 weeks; and correspondingly, 1.1924 Nm, 1.3092 Nm, and 1.3226 Nm for the MA implants. It can be concluded that the SLA surface achieves a better bone anchorage than the MA surface, and that sandblasting before acid etching has a beneficial effect on the interfacial shear strength. As regards the bone-implant interfacial stiffness calculated from the torque-rotation curve, the SLA implants showed an overall more than 5% higher stiffness compared with the MA implants, although the difference did not reach the statistical significance level. PMID- 11857441 TI - Surface characteristics and structure of anodic oxide films containing Ca and P on a titanium implant material. AB - An anodic oxide film that formed on titanium with a mixture of beta glycerophosphate sodium (beta-GP) and calcium acetate was investigated. The anodic oxide had interconnected pores (ca. 1-2 microm in diameter) and intermediate roughness (0.60-1.50 microm). In addition, it contained a mixture of amorphous, anatase, and rutile oxides. With an increase in the anodizing voltage and/or concentration of calcium incorporated into the oxide, the degree of oxide crystallinity increased. However, with an increase in the concentration of beta GP, the degree of oxide crystallinity decreased. It was concluded that the surface roughness, oxide crystallinity, and surface composition of the anodic oxide were dependent on the voltage, current density, and concentration of the electrolyte. It was also concluded that the anodized surface could be optimized for maximum osseointegration. PMID- 11857442 TI - Cripto: a tumor growth factor and more. AB - Cripto, a growth factor with an EGF-like domain, and the first member of the EGF CFC family of genes to be sequenced and characterized, contributes to deregulated growth of cancer cells. A role for Cripto in tumor development has been described in the human and the mouse. Members of the EGF-CFC family are found only in vertebrates: CFC proteins in zebrafish, Xenopus, chick, mouse and human have been characterized and indicate some common general functions in development. Cripto expression was first found in human and mouse embryonal carcinoma cells and male teratocarcinomas, and was demonstrated to be over-expressed in breast, cervical, ovarian, gastric, lung, colon, and pancreatic carcinomas in contrast to normal tissues where Cripto expression was invariably low or absent. Cripto may play a role in mammary tumorigenesis, since in vitro, Cripto induces mammary cell proliferation, reduces apoptosis, increases cell migration, and inhibits milk protein expression. This prediction is strengthened by observations of Cripto expression in 80% of human and mouse mammary tumors. At least three important roles for Cripto in development have created considerable interest, and each activity may be distinct in its mechanism of receptor signaling. One role is in the patterning of the anterior-posterior axis of the early embryo, a second is a crucial role in the development of the heart, and a third is in potentiating branching morphogenesis and modulating differentiation in the developing mammary gland. Whether these properties are functions of different forms of Cripto, different Cripto receptors or the distinct domains within this 15-38 kDa glycoprotein are examined here, but much remains to be revealed about this evolutionarily conserved gene product. Since all Cripto receptors have not yet been determined with certainty, future possible uses as therapeutic targets remain to be developed. Cripto is released or shed from expressing cells and may serve as an accessible marker gene in the early to mid-progressive stages of breast and other cancers. Meanwhile some speculations on possible receptor complexes for Cripto signaling in mammary cells are offered here as a spur to further discoveries. PMID- 11857443 TI - Many actions of cyclooxygenase-2 in cellular dynamics and in cancer. AB - Cyclooxygenase-2 (COX-2) is the inducible isoform of cyclooxygenase, the enzyme that catalyzes the rate-limiting step in prostaglandin synthesis from arachidonic acid. Various prostaglandins are produced in a cell type-specific manner, and they elicit cellular functions via signaling through G-protein coupled membrane receptors, and in some cases, through the nuclear receptor PPAR. COX-2 utilization of arachidonic acid also perturbs the level of intracellular free arachidonic acid and subsequently affects cellular functions. In a number of cell and animal models, induction of COX-2 has been shown to promote cell growth, inhibit apoptosis and enhance cell motility and adhesion. The mechanisms behind these multiple actions of COX-2 are largely unknown. Compelling evidence from genetic and clinical studies indicates that COX-2 upregulation is a key step in carcinogenesis. Overexpression of COX-2 is sufficient to cause tumorigenesis in animal models and inhibition of the COX-2 pathway results in reduction in tumor incidence and progression. Therefore, the potential for application of non steroidal anti-inflammatory drugs as well as the recently developed COX-2 specific inhibitors in cancer clinical practice has drawn tremendous attention in the past few years. Inhibition of COX-2 promises to be an effective approach in the prevention and treatment of cancer, especially colorectal cancer. PMID- 11857444 TI - Cdc7 kinase complex: a key regulator in the initiation of DNA replication. AB - DNA replication results from the action of a staged set of highly regulated processes. Among the stages of DNA replication, initiation is the key point at which all the G1 regulatory signals culminate. Cdc7 kinase is the critical regulator for the ultimate firing of the origins of initiation. Cdc7, originally identified in budding yeast and later in higher eukaryotes, forms a complex with a Dbf4-related regulatory subunit to generate an active kinase. Genetic evidence in mammals demonstrates essential roles for Cdc7 in mammalian DNA replication. Mini-chromosome maintenance protein (MCM) is the major physiological target of Cdc7. Genetic studies in yeasts indicate additional roles of Cdc7 in meiosis, checkpoint responses, maintenance of chromosome structures, and repair. The interplay between Cdc7 and Cdk, another kinase essential for the S phase, is also discussed. PMID- 11857445 TI - Role of N-cadherin in bone formation. AB - Cell-cell adhesion mediated by cadherins is essential for the function of bone forming cells during osteogenesis. Here, the evidence that N-cadherin is an important regulator of osteoblast differentiation and osteogenesis is reviewed. Osteoblasts express a limited number of cadherins, including the classic N cadherin. The expression profile of N-cadherin in osteoblasts during bone formation in vivo and in vitro suggests a role of this molecule in osteogenesis. Functional studies using neutralizing antibodies or antisense oligonucleotides indicate that N-cadherin is involved in the control the expression of osteoblast marker gene expression and differentiation. Cleavage of N-cadherin during osteoblast apoptosis also suggests a role of N-cadherin-mediated-cell-cell adhesion in osteoblast survival. Hormonal and local factors that regulate osteoblast function also regulate N-cadherin expression and subsequent cell-cell adhesion associated with osteoblast differentiation or survival. Signaling mechanisms involved in N-cadherin-mediated cell-cell adhesion and osteoblast gene expression have also been identified. Alterations of N-cadherin expression are associated with abnormal osteoblast differentiation and osteogenesis in pathological conditions. These findings indicate that N-cadherin plays a role in normal and pathological bone formation and provide some insight into the process involved in N-cadherin-mediated cell-cell adhesion and differentiation in osteoblasts. PMID- 11857446 TI - IGF-I mediates the stimulatory effect of high phosphate concentration on osteoblastic cell proliferation. AB - Although high concentrations of inorganic phosphate (Pi) are known to have a distinct anabolic effect on bone structure and metabolism, the precise mechanism by which phosphate possesses anabolic effect on bone formation has not been elucidated. The present study was performed to examine the effects of an increase in extracellular Pi concentration ([Pi](e)) on the proliferation of osteoblastic MC3T3-E1 cells. Increase in [Pi](e)(2-4 mM) dose-dependently stimulated DNA synthesis. Indomethacin, an inhibitor of prostaglandin synthesis, did not affect high [Pi](e)-induced DNA synthesis. DNA synthesis first increased affer a 3 h exposure to 4 mM [Pi](e) and its stimulatory effect was observed in a time dependent manner up to 24 h. On the other hand, DNA synthesis was significantly but partially blocked by cycloheximide, suggesting that this stimulatory effect of high [Pi](e) was at least in part dependent on new protein synthesis. There is recent evidence that MG3T3-E1 cells constitutively produce and secrete insulin like growth factor-I (IGF-I) and possess IGF-I receptors. IGF-I antiserum (1:10,000 to 1:100) significantly but partially blocked the stimulatory effect of [Pi](e) (4 mM) on DNA synthesis in a concentration-dependent manner. A neutralizing IGF-I antibody as well as IGF-I receptor antibody also significantly but partially blocked DNA synthesis stimulated by high [Pi](e) in a concentration dependent manner, indicating that IGF-I at least in part mediated the high [Pi](e)-induced effect. Actually, high [Pi](e) significantly increased the secretion of immunoreactive IGF-I into the medium as well as the expression of IGF-I mRNA. Present findings indicate that an increase in [Pi](e) stimulated DNA synthesis partly via an increase in IGF-I action. PMID- 11857447 TI - Effect of protein kinase inhibitors on the stretch-elicited c-Fos and c-Jun up regulation in human PDL osteoblast-like cells. AB - Osteoblastic cells transduce signals of mechanical loading that plays a key role in maintaining bone formation. In an attempt to elucidate the biochemical events associated with the conversion of mechanical stress to biological outcome, we examined cultured human periodontal ligament (hPDL) osteoblastic cells exposed to continuous stretch, in terms of cellular parameters correlating known signaling cascades to the initial phase of osteoblast-specific transcriptional control. Time-course experiments revealed that mechanical stretch-loaded hPDL cells exhibit a very rapid and relatively sustained increase in the abundance of the immediate-early gene products, c-Fos and c-Jun, components of the activator protein-1 (AP-1) transcription factor. Moreover, this increase in protein levels was accompanied by hyperphosphorylation and thereby potentiation of c-Jun, the principal modulator of AP-1 activity. Importantly, these inductive effects were partly or completely abolished by pre-incubating the cells with SB 203580, PD 098059, and the novel compound Y-27632, inhibitors of p38 mitogen-activated protein kinase (MAPK), MAPK kinase (MEK), and Rho-associated protein kinase (RhoK), respectively. These results consolidate AP-1 as the pivotal downstream effector in the early response of hPDL cells to continuous mechanical stretching, via the coordinate stimulation of de novo synthesis and post-translational regulation of AP-1 proteins. This "integrating" function of AP-1 is mediated through a mechanotransduction circuit that incorporates elements of well-defined upstream signaling protein kinase systems. PMID- 11857449 TI - Termination of lifespan of SV40-transformed human fibroblasts in crisis is due to apoptosis. AB - Normal human fibroblasts in culture have a limited lifespan, ending in replicative senescence. Introduction of SV40 sequences encoding large T antigen and small t antigen into pre-senescent cells results in an extension of lifespan for an additional 20-30 population doublings. Rare clones of SV40-transformed cells are capable of indefinite growth and are described as immortal; however, the great majority of SV40-transformed cells terminate this extended lifespan in cell death, termed "crisis." We have examined the properties of cells in crisis to obtain further insights into mechanism of cell death and immortalization. Populations at the terminal cell passage show a balance between cell replication and cell death over a period of several weeks, with a progressive increase in cells undergoing cell death. During this period, there is less than a 3-fold increase in attached cell number, with two stages being identifiable on the basis of the focal pattern of cell survival. We also demonstrate that cells in crisis are undergoing apoptosis based on TUNEL assay, subG1 DNA content, annexin V reactivity, and activation of caspases 3 and 8. We suggest a model whereby SV40 transformed cells acquire increased sensitivity to apoptosis based on changes in properties which activate caspase 8 in addition to changes previously described involving shortening of telomeric sequences. While only telomere stabilization could be clearly shown to be essential for survival of cells through crisis, the extended period of cell replication and altered gene expression observed in SV40 transformed cells during crisis are compatible with other genetic alterations in immortal cells. PMID- 11857448 TI - Osteogenic protein-1 and interleukin-6 with its soluble receptor synergistically stimulate rat osteoblastic cell differentiation. AB - Osteogenic Protein-1 (OP-1, BMP-7), a member of the bone morphogenetic protein family, stimulates synthesis of biochemical markers characteristic of the osteoblastic and chondrocytic phenotypes and induces new bone formation. Interleukin-6 (IL-6), a cytokine produced by a wide variety of cells, appears to interact with other factors producing different biological effects. In the present study, we showed that OP-1 action in fetal rat calvaria (FRC) cells was enhanced by the combination of IL-6 and the soluble receptor IL-6sR. OP-1 alone induced alkaline phosphatase (AP) activity by 4- to 5-fold above the control. Exogenous IL-6 soluble receptor (IL-6sR) synergistically stimulated the OP-1 induced AP activity and mineralized bone nodule formation by an additional 3 fold. The stimulation was IL-6sR concentration-dependent. The combination of IL-6 and IL-6sR synergistically stimulated OP-1 action by an additional 6- to 7-fold. BMPR-II receptor mRNA expression in FRC cells treated with OP-1 and IL-6 plus IL 6sR was stimulated further, while BMPR-IA, -IB, and ActR-I expressions were not affected. The intracellular signaling molecules Smad2 and Smad5 mRNA expressions were not changed under these conditions. The expression of selected BMP family members (BMP-3, -4, and -6) was altered in FRC cells treated with OP-1 in combination with IL-6 and IL-6sR. The combination of IL-6 and IL-6sR reduced the OP-1-stimulated BMP-3 mRNA levels and enhanced the suppressive effect of OP-1 on BMP-4 and -6 mRNA expressions. In conclusion, the present results demonstrate that exogenous IL-6 and IL-6sR synergistically stimulate OP-1 action in primary cultures of rat osteoblastic cells. One possible mechanism of synergy involves differential regulation of the effects of OP-1 on the expression of the type II BMP receptor and several other BMPs. PMID- 11857450 TI - Profiling of retinoid mediated gene expression in synchronized human SCC cells using Atlas human cDNA expression arrays. AB - While retinoids have been demonstrated to inhibit growth of many tumor cells, including SCC cells, the molecular mechanism by which retinoids suppress growth has not been elucidated. We previously found that the growth of SCC cells was significantly inhibited by all-trans-retinoic acid (all-trans-RA) treatment, and this inhibition was dependent on the binding and activation of RARs. These nuclear receptors bind retinoids and alter the rate of transcription of specific genes. To identify targets of the activated RARs which mediate growth inhibition, we growth arrested SCC-25 cells in G-0 and examined the effect of all-trans-RA on synchronized SCC-25 cells. All-trans-RA inhibited G-1 progression in quiescent SCC-25 cells stimulated by FBS. More specifically, we found that the all-trans-RA execution point maps to mid/late G-1, 6 to 10 h after stimulation. Using this synchronized cell system, we examined the expression of cell cycle regulatory genes in quiescent SCC-25 cells stimulated with FBS and treated with all-trans RA. We found few changes in expression of these genes which could account for all trans-RA inhibition of SCC-25 cell growth. In order to compare the patterns of expression of a wider selection of genes in all-trans-RA treated and non-treated SCC-25 cells, we have used expression array technology. We successfully performed expression profiling experiments on the Atlas Human cDNA arrays which contain 1176 human genes. We have identified several up-regulated and several down regulated gene expression changes mediated by all-trans-RA treatment in synchronized SCC-25 cells. This novel information will be useful in defining the mechanism by which retinoids suppress the growth of SCC cells. PMID- 11857451 TI - EGFR-independent activation of ERK1/2 mediates growth inhibition by a PTPase antagonizing K-vitamin analog. AB - The K-vitamin analog Cpd 5 or [2-(2-mercaptoethanol)-3-methyl-1,4-napthoquinone] is a potent cell growth inhibitor in vitro and in vivo, likely due to arylation of enzymes containing a catalytic cysteine. This results in inhibition of protein tyrosine phosphatase (PTPase) activity with resultant hyperphosphorylation of EGF receptors (EGFR) and ERK1/2 protein kinases, which are downstream to EGFR in the MAPK pathway. We used NR6 fibroblast cells, which lack endogenous EGFR and its variant cells transfected with different EGFR mutants to assess the contribution of the EGFR-mediated signaling pathway to Cpd 5-mediated ERK activation and cell growth inhibition. Cpd 5 treatment resulted in enhanced phosphorylation of EGFR at carboxyl-terminal tyrosines. This phosphorylation and activation of EGFR were found to be necessary neither for growth inhibition nor for the activation of the downstream kinases ERK1/2, since both occurred in EGFR-devoid mutant cells. U0126 and PD 098059, specific inhibitors of MEK1/2, the ERK1/2 kinases, antagonized both cell growth inhibition and ERK1/2 phosphorylation mediated by Cpd5. Cpd 5 was also found to inhibit ERK1/2 phosphatase(s) activity in lysates from all the cells tested, irrespective of their EGFR status. These results show that EGFR independent ERK1/2 phosphorylation was involved in the mechanism of Cpd5 mediated growth inhibition. This is likely due to the observed antagonism of ERK phosphatase activity. A candidate PTPase was found to be Cdc25A, a recently identified ERK phosphatase. PMID- 11857452 TI - Effects of hyperthermia on p53 protein expression and activity. AB - Although p53 responses after DNA damage have been investigated extensively, p53 responses after heat shock, which exerts cytotoxic action by mechanisms other than direct induction of DNA damage, are less well characterized. We investigated, therefore, the effect of hyperthermic exposures on the levels and DNA-binding activity of p53. Experiments were carried out with U2OS and ML-1 cells, known to express wild-type p53 protein. Although heating at 41 degrees C for up to 6 h had only a small effect on p53 levels or DNA binding activity, exposure to temperatures between 42.5 and 45.5 degrees C caused an immediate decrease in protein levels that was associated with a reduction in DNA binding activity. This observation is compatible with a high lability of p53 to heat shock, or heat sensitivity of the pathway regulating p53 levels in non-stressed cells. When cells were heated to 42.5 degrees C and returned to normal temperatures, a strong p53 response associated with an increase in protein levels and DNA binding activity was observed, suggesting the production of p53-inducing cellular damage. At higher temperatures, however, this response was compromised in an exposure-time-dependent manner. The increase in DNA binding activity was more heat sensitive than the increase in p53 levels and was inhibited at lower temperatures and shorter exposure times. Thus, the pathway of p53 activation is itself heat sensitive and compromised at high levels of exposure. Compared to p53 activation after exposure to ionizing radiation, heat-induced activation is rapid and short lived. When cells were exposed to combined heat and radiation, the response observed approximated that of cells exposed to heat alone. Wortmannin at 10 microM inhibited p53 activation for up to 2 h after heat shock suggesting the involvement of wortmannin-sensitive kinases, such as DNA-PK and ATM. Heat shock causes phosphorylation of p53 at Serine-15, but there is no correlation between phosphorylation at this site and activation of the protein. The results in aggregate indicate p53 activation in the absence of DNA damage by a heat sensitive mechanism operating with faster kinetics than radiation-induced p53 activation. The former response may induce pathways preventing other stimuli from activating p53, as heat-induced activation of p53 is dominant over activation of p53 by DNA damage in combined-treatment experiments. These observations suggest means for abrogating p53 induction after DNA damage with the purpose of potentiating response and enhancing cell killing. PMID- 11857453 TI - Transforming growth factor-beta isoforms differently stimulate proalpha2 (I) collagen gene expression during wound healing process in transgenic mice. AB - The role of many growth factors and cytokines in the process of wound healing has been intensively investigated in recent two decades. Among them, transforming growth factor-betas (TGF-betas) are well known to have a potent stimulatory effect on collagen synthesis as shown in various in vivo experimental systems. In the present study, we examined the effects of various growth factors on the promoter activity of the proalpha2 (I) collagen gene (COL1A2) during the wound healing process. For this purpose, we utilized transgenic mice harboring the -17 kb promoter sequence of the mouse COL1A2 linked to either a firefly luciferase or a bacterial beta-galactosidase gene. These mice exhibited normal phenotypic expression and the wound healing process was not impaired. Full thickness wounds were made by punch biopsy. We examined the effects of TGF-beta1, -beta2, -beta3, basic fibroblast growth factor, platelet-derived growth factor, and connective tissue growth factor by applying them locally to the open wound every 2 days. Among the growth factors examined, all of the three isoforms of TGF- exhibited a more potent stimulatory effect on COL1A2 promoter activity than did other factors. In addition, while TGF-beta1 and -beta2 significantly increased the number of fibroblasts which were positive for X-Gal staining, TGF-beta3 treatment did not change the number of beta-galactosidase expressing cells. Accumulation of collagen fibers was observed to the same extent in the mice treated with TGF beta1 and those with TGF-beta3. These findings suggest that TGF-beta1 and -beta3 have similar but not identical regulatory mechanisms of COL1A2 expression, and that their pathophysiological roles in wound healing might be different from each other. PMID- 11857455 TI - Generating new marine cell lines and transgenic species--conference summary. AB - Marine species offer a tremendous diversity of life histories, physiologies, genetics, behaviors, and biologies, reflecting myriad adaptations to the water environment. Historically, marine vertebrates, particularly fish, have played significant roles in a wide range of disciplines, including environmental toxicology, genetics, developmental biology, and physiology, among others. Much still remains to be learned from these animals, and there is a growing need for new marine models. Models for expression of marine animal genes have been limited to heterologous expression systems. While there is still a great deal to gain from heterologous expression systems, the interactions of genes with one another can best be determined in homologous expression systems where appropriate interactions are possible. This has become particularly important with the development of functional genomics in marine models. These homologous gene expression systems will be key to the use of functional genomics for marine animal molecular physiology and toxicology. PMID- 11857454 TI - Acute endotoxemia prolongs the survival of rat lung neutrophils in response to 12 O-tetradecanoyl-phorbol 13-acetate. AB - Acute endotoxemia is associated with prolonged survival of adherent neutrophils in the lung vasculature. In the present studies, the effects of inflammatory mediators on signaling pathways regulating neutrophil survival were examined. We found that the protein kinase C activator, 12-O-tetradecanoyl-phorbol 13-acetate (TPA), but not interferon-gamma (IFN-gamma), prolonged the survival of adherent vasculature lung neutrophils from endotoxemic rats, a response that was correlated with reduced apoptosis. Although endotoxin administration to rats induced the expression of the anti-apoptotic protein Mcl-1 in lung neutrophils, TPA had no effect on this response. Endotoxin administration also induced expression of total p38 and p44/42 mitogen activated protein kinases (MAPK) in neutrophils, as well as phosphatidyl inositol 3 kinase (PI3K) and its downstream target protein kinase B (PKB). Treatment of the cells with TPA increased p38 MAPK expression in cells from both control and endotoxin treated animals. Cells from endotoxin treated, but not control animals, were found to exhibit constitutive binding activity of nuclear factor kappa B (NF-kappaB) which was blocked by TPA. In contrast, constitutive CCAAT/enhancer binding protein (C/EBP) nuclear binding activity evident in neutrophils from control animals was reduced following endotoxin administration. Moreover, this response was independent of TPA. These data suggest that NF-kappaB plays a role in TPA-induced signaling leading to prolonged survival of adherent vascular neutrophils in the lung during acute endotoxemia. PMID- 11857456 TI - Effects of plasma from bivalve mollusk species on the in vitro proliferation of the protistan parasite Perkinsus marinus. AB - The in vitro culture of the Eastern oyster parasite Perkinsus marinus has provided a unique opportunity to examine its susceptibility to putative recognition and effector defense mechanisms operative in refractory bivalve species. In this study, we report the effect of supplementing the culture medium with plasma from: (1) uninfected to heavily infected Eastern oysters; (2) oyster species considered to be disease-resistant; and (3) bivalve mollusk species that are naturally exposed to the parasite but show no signs of disease. We also examined in vitro the interaction between hemocytes from Crassostrea virginica and C. gigas and P. marinus trophozoites. Our results revealed a significant decrease (32%) in proliferation of P. marinus in the presence of plasma from heavily infected C. virginica oysters. The inhibitory effects were less pronounced with plasma from moderately infected and uninfected oysters. In contrast, plasma from C. rivularis and C. gigas enhanced P. marinus proliferation. Proliferation was significantly reduced in media supplemented with plasma from Mytilus edulis, Mercenaria mercenaria, and Anadara ovalis. The highest inhibitory activity was apparent in M. edulis, for which 5% plasma supplemented medium reduced growth by 35% relative to the controls. M. edulis active component(s) was heat-stable, yet pronase-sensitive. The significantly higher uptake of live P. marinus trophozoites by hemocytes from C. virginica, relative to those from C. gigas, suggests a certain level of specificity in the recognition/endocytosis of the parasite by its natural bivalve host species. PMID- 11857457 TI - Cold acclimation induces proliferation of sarcoplasmic reticulum without increase in Ca2+-ATPase activity in white axial muscle of striped bass (Morone saxatilis). AB - The effects of acclimation of striped bass to cold (5 degrees C) and warm (25 degrees C) temperatures upon ultrastructural features of white axial skeletal muscle are quantified. Surface density of sarcoplasmic reticulum (SR) increased by almost 30%, and SR volume density increased by about 20% during cold acclimation. Proliferation of SR suggests an increase in available SR surface for re-sequestration of Ca2+ and a decrease in diffusion path length for Ca2+ during cold acclimation. Average cross-sectional areas and cross-sectional perimeters of myofibrils situated in the center of muscle fibers decreased during cold acclimation by approximately 20% and 11%, respectively. Additionally, average major and minor axes of ellipses fit to central myofibrillar cross-sections decreased by approximately 12% and 8%, respectively, during cold acclimation. These measurements define a decrease in average myofibrillar diameter and suggest a decrease in diffusion path length for Ca2+ to and from myofibrillar activation sites. Measurements of peripheral myofibrils that had elongated profiles in cross sections indicate that maximum profile length of these myofibrils decreases by about 17%. Peripheral myofibrils may break up into smaller myofibrils with more rounded cross-sectional profiles during cold acclimation. SR Ca2+-ATPase of white axial muscle was also measured in unfractionated homogenates and in crude SR enriched subcellular fractions from cold- and warm-acclimated striped bass. No difference in SR Ca2+-ATPase activity per g wet weight was observed between cold- and warm-acclimated animals. Lack of increase in SR Ca2+-ATPase per g wet weight, despite a significant proliferation of SR, probably results in a decrease in average Ca2+-ATPase pump density within the SR membrane during cold acclimation. Thus, compensation for decreased diffusion coefficient of Ca2+ during cold acclimation appears due to the combined effects of proliferation of SR surface density and a decrease in average myofibrillar diameter. PMID- 11857458 TI - Capillary plexus development in the day five to day six chick chorioallantoic membrane is inhibited by cytochalasin D and suramin. AB - The chick chorioallantoic membrane (CAM) is a valuable model for evaluating angiogenesis and vasculogenesis. Our purpose was to characterize the formation of the CAM vasculature, in particular the capillary plexus, between days five and six after fertilization and to examine the mode of action of cytochalasin D and suramin on vascular development during this interval. The CAM increased 20-fold in size between days five and six, during which time the capillary plexus forms by both migration of mesodermal blood vessels toward the ectoderm and by the formation of new vessels from angioblasts near the ectoderm. Between days five and six, the CAM becomes thinner, and the density of the mesodermal cells decreases. To determine the mode of action of anti-angiogenic drugs on the day five to day six CAM, various concentrations of cytochalasin D or suramin were added directly to day five CAMs, and their effects were evaluated on day six. Both drugs significantly inhibited CAM growth, altered branching patterns of the major vessels, decreased area of the major vessels, and inhibited the formation of the capillary plexus by inhibiting both vasculogenesis and the migration of mesodermal blood vessels to the ectoderm. Cytochalasin D also inhibited compartmentalization of the plexus. Cytochalasin D and suramin were inhibitory at similar doses. This study provides new information on early CAM development, establishes the mode of action and dose dependency of cytochalasin D and suramin on day five to day six CAMs, and demonstrates that the day five to day six CAM provides a useful assay to examine the effect of anti-angiogenic drugs on blood vessel development, including capillary plexus formation. PMID- 11857459 TI - Vasculature in pre-blastema and nerve-dependent blastema stages of regenerating forelimbs of the adult newt, Notophthalmus viridescens. AB - Immunocytochemistry utilizing a monoclonal antibody (BV1; blood vessel 1) highly reactive to the vasculature of the adult newt showed that a developing vasculature was present during early, pre-blastema, and early-bud blastema stages of forelimb regeneration in this species. Infusion of Prussian Blue and DiI into the brachial artery further delineated the intactness of this early vasculature. Finally, macroscopic observations of vascular flow underneath the apical epithelial cap (AEC) and microsurgical removal of the AEC and observation of subsequent bleeding buttressed the conclusion that an intact vasculature exists during early nerve-dependent stages of newt forelimb regeneration. The results suggest that this process of neovascular formation is angiogenesis, i.e., the formation of new vessels from pre-existing vessels in the stump. Furthermore, angiogenesis is an ongoing process initiated early after amputation. Blastema cells and the AEC are likely sourcesof factors that stimulate neovascularization. PMID- 11857460 TI - Presence of two neuropeptides in the fusiform ganglion and reproductive ducts of Octopus vulgaris: FMRFamide and gonadotropin-releasing hormone (GnRH). AB - We have found evidence of FMRFamide-like and cGnRH-I-like immunoreactivity in the central nervous system (CNS) and in the reproductive ducts of both female and male cephalopod Octopus vulgaris. Cell bodies and fibers were immunolocalized in the fusiform ganglion from which the nerves that reach the female and male reproductive ducts arise. FMRFamide-like and cGnRH-I-like immunoreactive nerve endings were present in the oviduct, and in the oviducal gland of the female and in the seminal vesicle of the male. The GnRH-like peptide from the reproductive ducts has been partially characterized by HPLC. The retention time of the Octopus vulgaris GnRH-like peptide was similar to the retention time of cGnRH-I. Based on these observations we suggest that FMRFamide-like and a novel GnRH-like peptide are involved in the control of reproductive ducts of Octopus vulgaris. One possibility is that the peptides affect gamete transport. Another possibility is that they regulate secretory products such as mucus and mucilaginous substances from the oviducal gland and the seminal vesicle. Our data provide further evidence to support the hypothesis of the existence of a central and peripheral peptidergic control of reproduction of Octopus vulgaris. PMID- 11857461 TI - Role of melatonin in photoperiodic time measurement in the migratory redheaded bunting (Emberiza bruniceps) and the nonmigratory Indian weaver bird (Ploceus philippinus). AB - In the present study, we asked the question whether physiological responses to day length of migratory redheaded bunting (Emberiza bruniceps) and nonmigratory Indian weaver bird (Ploceus philippinus) are mediated by the daily rhythm of melatonin. Melatonin was given either by injection at certain times of the day or as an implant. In series I experiments on the redheaded bunting, melatonin was administered by subcutaneous injections daily at zeitgeber time (ZT) 4 (morning) or ZT10 (evening) and by silastic capsules in photosensitive unstimulated buntings that were held in natural day lengths (NDL) at 27 degrees N beginning from mid February, and in artificial day lengths (ADL, 12L:12D and 14L:10D). Melatonin did not affect the photoperiod-induced cycles of gain and loss in body mass and testicular growth-involution, but there was an effect on temporal phasing of the growth-involution cycle of testes in some groups. For example, the rate of testicular growth and development was faster in birds that received melatonin injection at ZT4 in NDL, and was slower in birds that carried melatonin implants both in NDL and ADL. In series II experiments on Indian weaver birds, melatonin was given in silastic capsules in the first week of September when they still had large gonads. Birds were exposed for 12 weeks to short day length (8L:16D; group 1), to long day length (eight weeks of 16L:8D and four weeks of 18L:6D; group 2), or to both short and long day lengths (four weeks each of 8L:16D, 16L:8D, and 18L:6D; groups 3 and 4). Whereas groups 1 to 3 carried melatonin or empty implant from the beginning, group 4 received one after four weeks. All birds underwent testicular regression during the first four weeks irrespective of the photoperiod they were exposed to or the implant they carried in, and there was a slight re-initiation of testis growth in some birds during the next eight weeks of long day lengths. However, with the exception of group 2, there was no difference in mean testis volume during the period of experiment between the melatonin- and empty-implant birds. The data on androgen-dependent beak color also supported the observations on testes. Together, these results do not support the idea that the daily rhythm of melatonin is involved in the photoperiodic time measurement in birds. However, there may still be a role of melatonin in temporal phasing of the annual reproductive cycle in birds. PMID- 11857462 TI - Environmental antiecdysteroids alter embryo development in the crustacean Daphnia magna. AB - The role of ecdysteroids in crustacean embryo development and the susceptibility of the developing embryo to the antiecdysteroidal properties of an environmental chemical were evaluated. The agricultural fungicide fenarimol was shown to exhibit antiecdysteroidal activity to the crustacean Daphnia magna by lowering endogenous ecdysone levels and delaying molting in a concentration-dependent fashion that was mitigated by co-exposure to exogenous 20-hydroxyecdysone. Exposure of either gravid maternal organisms or isolated embryos to fenarimol resulted in embryo abnormalities ranging from early partial developmental arrest to incomplete development of antennae and shell spines. Developmental abnormalities were associated with suppressed ecdysone levels in the embryos and the abnormalities could be prevented by co-exposure to 20-hydroxyecdysone. Developmental abnormalities caused by the antiecdysteroid were associated with reduced fecundity of the parental organisms. These results demonstrate that ecdysteroids are critical to normal crustacean embryo development and environmental antiecdysteroids can disrupt normal embryo development and compromise the production of viable offspring. Antiecdysteroidal activity may provide a means by which environmental chemicals impact crustacean species while not affecting vertebrates. PMID- 11857463 TI - Catecholamines are present in larval Xenopus laevis: a potential source for cardiac control. AB - Changes in noradrenaline (NA), adrenaline (A), and dopamine (DA) levels in the heart, kidneys, and whole body (without heart and kidneys) during embryonic development were investigated in the frog, Xenopus laevis using high-performance liquid chromatography (HPLC). In addition, the presence of cells immunoreactive to tyrosine hydroxylase (TH), dopamine-beta-hydroxylase (DBH) and/or phenylethanolamine-N-methyltransferase (PNMT) in the heart of Xenopus larvae was investigated using immunohistochemical techniques. The presence of nerve fibers was visualized using antibodies against acetylated tubulin (AcT). NA and DA concentrations in the heart were low and steady in NF 40-56, showed an increased value at NF 57, and decreased again in froglets. A trend toward higher concentrations of A was observed at NF 43-49 and NF 57. Cells immunoreactive to TH, DBH, and PNMT were found in the heart from NF 40, and the TH immunoreactive cells became more abundant in the whole heart at later stages. The presence of catecholamines in the non-innervated larval heart together with the finding of TH/DBH/PNMT immunoreactive cells suggests that catecholamines are synthesized and stored in the heart and could therefore have a paracrine role in cardiac control in Xenopus larvae. Detectable concentrations of catecholamines were also found in kidneys and whole bodies (except heart and kidneys). Therefore, catecholamine producing cells outside the heart can be an important source of circulating catecholamines involved in adrenergic cardiac control in Xenopus larvae. PMID- 11857464 TI - Basic properties and annual changes of follicle-stimulating hormone receptors in the testis of horseshoe bats, Rhinolophus ferrumequinum. AB - The unique reproductive patterns, delayed fertilization in females, and asynchrony between spermatogenesis and mating behavior in males are well documented in bats living in temperate latitudes. The present study was undertaken to examine follicle-stimulating hormone (FSH) receptors in the testis of bats, Rhinolophus ferrumequinum, during the annual reproductive cycle. Male bats were captured at natural roosting sites and testicular preparations were subjected to a radioligand binding assay for FSH receptors. The weight of paired testes increased considerably in the spermatogenic period and decreased from the mating to hibernation periods. Meiotic division in the testis was observed in the spermatogenic period but not the mating period. Serum testosterone concentrations increased in the spermatogenic period and rapidly decreased in the mating period. The binding of FSH was specific for mammalian FSHs and detected primarily in the testis. Scatchard plot analyses of the binding of FSH to bat testicular preparations showed straight lines, suggesting the presence of a single class of binding sites. The affinities (equilibrium association constant) of FSH receptors were consistent throughout the annual reproductive cycle. The specific binding per unit weight of testis and total binding in the paired testes were highest in the mating period and in the spermatogenic period, respectively, among reproductive periods. The accumulation of cyclic adenosine 3', 5'-monophosphate to FSH stimulation was higher in the spermatogenic period than in the hibernation period. These findings suggest that testicular function of bats is associated with seasonal changes in the number of binding sites, while the number per target cell and the activation of adenylate cyclase led by FSH-receptor complex considerably decreases in the hibernation period. PMID- 11857465 TI - Egg-jelly structure promotes efficiency of internal fertilization in the newt, Cynops pyrrhogaster. AB - Egg-jelly is composed of a network of fibrous components and contains substances regulating the sperm-egg interaction. Many studies on the latter have been conducted, whereas the role of the egg-jelly structure in fertilization has not yet been fully assessed. In this study, we examined the fertilization efficiency in the presence and absence of the structure around the egg of the newt, Cynops pyrrhogaster, using a gelatin gel system. Although de-jellied eggs of C. pyrrhogaster can be fertilized with an adequate number of sperm, the fertilization rate was dramatically increased through the use of the gelatin gel. Sperm showed forward motility with straight morphology in the gel, whereas they swam in circles in solution. This result indicates that the gel structure is significant for sperm guidance to the egg surface, and its presence raises the fertilization efficiency in C. pyrrhogaster. When sperm were entangled in the gel structure, they were immediately folded and never showed any forward motility. Sperm with zigzag morphology were observed in the gelatin gel as well as in the egg-jelly, indicating the elimination of sperm by the gel structure. The effect of sperm elimination on successful fertilization was estimated using gelatin gels of different thickness. Though the variation did not affect the fertilization rate, the rate of normal development gradually increased in the thicker gels. This result indicates that sperm elimination in egg-jelly can function in the fertilization system. The roles of sperm guidance and sperm elimination under the physiological condition of internal fertilization of the newt are discussed. PMID- 11857466 TI - Synthesis, distribution, and levels of an egg lipoprotein from the apple snail Pomacea canaliculata (Mollusca: Gastropoda). AB - The site of synthesis of mollusc lipoproteins is hitherto unknown and was investigated for perivitellin 2 (PV2), an egg lipoprotein found in the freshwater snail Pomacea canaliculata. Tissues (albumen gland, gonad-digestive gland complex, and muscle) from vitellogenic females were incubated in vitro with 14C leucine at 25 degrees C for 12 hr. At the end of incubation, soluble proteins from tissue homogenates and medium were analyzed for de novo protein synthesis by electrophoresis and HPLC, and radiolabeled proteins were quantified by liquid scintillation. Two albumen gland radiolabeled proteins (67 and 31 kDa) co migrated with the subunits of PV2, and they represented 6.0% of the total labeled protein in that tissue. Western blot analysis confirmed the presence of PV2 only in the albumen gland. In vivo experiments where adult females were injected with 3H-leucine revealed that PV2 was not present in hemolymph. ELISA analysis in all tissues of the snail confirmed the PV2 presence only in the albumen gland and developing eggs with levels of 26 and 98 mg/g protein, respectively. Therefore, the albumen gland is the only site for PV2 synthesis, and no extra-gland synthesis, circulation, or accumulation could be found. PV2 subunits were further characterized analyzing N-terminal sequences which showed no homology with other proteins. PMID- 11857467 TI - Histidine uptake in the epidermis of lizards and snakes in relation to the formation of the shedding complex. AB - Mammalian epidermis utilizes histidine-rich proteins (filaggrins) to aggregate keratin filaments and form the stratum corneum. Little is known about the involvement of histidine-rich proteins during reptilian keratinization. The formation of the shedding complex in the epidermis of snakes and lizards, made of the clear and the oberhautchen layers, determines the cyclical epidermal sloughing. Differently from snakes, keratohyalin-like granules are present in the clear layer of lizards. The uptake of tritiated histidine into the epidermis of two lizards and one snake has been studied by autoradiography in sections at progressive post-injection periods. At 40 min and 1 hr post-injection keratohyalin-like granules were not or poorly labeled. At 3-22 hr post-injection most of the labeling was present over suprabasal cells destined to form the shedding complex, in keratohyalin-like granules of the clear layer, and in the forming a-layer but was low in the forming b-layer, and in superficial keratinized layers. The analysis of the shedding complex in the pad lamellae (a specialized scale used for climbing) of a gecko showed that the setae and the cytoplasm of clear cells among them are main sites of histidine uptake at 4 hr post-injection. In the snake most of the labeling at 4 hr post-injection was localized in the shedding complex along the boundary between the clear and oberhautchen layers. The present study suggests that, in the epidermis of lepidosaurian reptiles, the synthesis of a histidine-rich protein is involved in the formation of the shedding layer and, as in mammals, in a-keratinization. PMID- 11857468 TI - Energy and water conservation in frozen vs. supercooled larvae of the goldenrod gall fly, Eurosta solidaginis (fitch) (Diptera: Tephritidae). AB - Insects that tolerate severe cold during winter may either supercool or tolerate ice forming within the tissues of the body. To compare the relative advantages of freezing and supercooling, we measured rates of CO(2) production and water loss in frozen and supercooled goldenrod gall fly larvae (Eurosta solidaginis). As an important first step, we measured the time required for ice content and metabolic rate to stabilize upon freezing. Ice content stabilized after only three hours of freezing at -5 degrees C, whereas CO(2) production required 12 hours to stabilize. Subsequent experiments found that freezing greatly reduced both water loss and metabolic rate. Comparisons of supercooled and frozen larvae at -5 degrees C indicated that CO(2) production fell 47% with freezing and water loss decreased 35%. As temperature decreased to -10 and -15 degrees C, CO(2) production fell exponentially and was no longer detectable at -20 degrees C with our measurement system. Our results demonstrate that freezing significantly reduces energy consumption during the winter and may therefore improve winter survival and spring fecundity. The advantages of freezing over supercooling would drive selection toward insect freeze tolerance and also toward higher supercooling points to increase the duration of freezing each winter. PMID- 11857469 TI - Effects of genic substitution at the pink-eyed dilution locus on the proliferation and differentiation of mouse epidermal melanocytes in vivo and in vitro. AB - Cells positive to the dopa reaction (melanocytes) as well as to the combined dopa premelanin reaction (melanoblasts and melanocytes) in the epidermis of C57BL/10JHir-p/p (pink-eyed dilution) mice were fewer and less reactive than in C57BL/10JHir (black, P/P) mice, suggesting that the proliferation and differentiation of p/p melanocytes are inhibited. To confirm the inhibitory effects of p gene on the proliferation and differentiation of epidermal melanocytes, we cultured epidermal cell suspensions of neonatal skins from P/P and p/p in a serum-free medium. The proliferation and differentiation of p/p melanoblasts/melanocytes in primary culture were greatly inhibited as compared to P/P melanoblasts/melanocytes. The morphology of p/p melanoblasts/melanocytes cultured in melanocyte growth medium, though non-pigmented, was similar to P/P melanocytes; namely, dendritic, polygonal, or epithelioid. About 8% of p/p cells cultured in melanocyte growth medium were positive to the dopa reaction, and about 25% were reactive to the combined dopa-premelanin reaction. Eumelanin content in p/p was extremely reduced compared to P/P. The immunocytochemical staining of p/p melanoblasts/melanocytes revealed that they are negative to tyrosinase, but reactive to tyrosinase-related protein (TRP)-1, TRP-2, and c-kit. However, the reactivities in p/p were lower than in P/P. Although the differentiation of p/p melanoblasts was not induced by endothelin (ET)-1, ET-2, and ET-3, the proliferation of p/p melanoblasts was stimulated by them. These results suggest for the first time that p gene exerts its influence on the proliferative activities of mouse epidermal melanoblasts by affecting the regulatory mechanisms dependent on the function of ETs. PMID- 11857470 TI - Characterization and quantification of yolk proteins in the lobster, Homarus americanus. AB - Yolk protein (vitellin, Vn) and its precursor (vitellogenin, Vg) were isolated and characterized in the ovary and hemolymph, respectively, of the adult female lobster, Homarus americanus. Vn had a molecular mass of 360 kDa when analyzed by gel filtration. When analyzed by SDS-PAGE, Vn had six bands (110, 105, 94, 90, 81, and 78 kDa). An anti-Vn antiserum was developed using purified Vn, and the antiserum was used to detect Vn and Vg by ELISA and western blot techniques. ELISA analysis of hemolymph proteins separated by gel filtration indicated that Vg was similar in mass to Vn (360 kDa). However, western blots of hemolymph proteins separated by SDS-PAGE indicated that Vg contained a pair of protein subunits, 194 kDa and 179 kDa. Furthermore, the elution profiles of Vn and Vg from anion exchange chromatography indicated that Vg had a more negative charge. Thus, Vg appears to be processed after its uptake by the ovary to form Vn. Vg was undetectable in hemolymph from adult males by either ELISA or by western blot analysis. However, hemolymph levels of Vg in adult females increased 40-fold during the reproductive cycle, rising from 18 microg/mL in ovarian stage II to 789 microg/mL at stage V. This increase correlates well with oocyte growth during the cycle. Hence, this method may be useful for studying the regulation of lobster vitellogenesis. PMID- 11857471 TI - Selective enhanced phosphorylation of shrimp beta-tubulin by PKC-delta with PEP(taxol), a synthetic peptide encoding the taxol binding region. AB - Beta-tubulin cDNA from the shrimp Penaeus japonicus was isolated by homology cloning. Expression of cDNA in Escherichia coli yielded a 55 kDa polypeptide, positive for monoclonal antibodies against mammalian beta-tubulin. Autoradiography demonstrated the bacterially expressed hepatopancreas beta tubulin of P. japonicus is specifically phosphorylated by the delta isoenzyme of protein kinase C (PKC-delta) purified from the plasma membrane of the shrimp heart, in the presence of the receptor for activated PKC (RACK), but not in its absence. Purified shrimp heart PKC-delta is able to phosphorylate bacterially expressed shrimp beta-tubulin without the presence of Ca(++), but requires Mg(++). The kinase activity of purified PKC-delta on bacterially expressed beta tubulin was enhanced by incubation with PEP(taxol), a synthetic peptide encoding the taxol-binding region of beta-tubulin. In other words, PEP(taxol) modulates the kinase activity of PKC-delta through RACK. PMID- 11857472 TI - Seasonal changes in circulating steroid concentration and their correlation with the ovarian activity in the female Indian sheath-tailed bat, Taphozous longimanus. AB - The relationship between ovarian activity and circulating steroid concentration was studied in the female sheath-tailed bat, Taphozous longimanus. T. longimanus breeds twice in rapid succession during the year at Varanasi, India. Ovarian recrudescence was observed during September, and antral follicles were first observed during the month of October. Circulating androstenedione concentration showed an increase beginning in October, reaching a peak in December. This increase in androstenedione concentration correlated with the period of heavy accumulation of adipose tissue and increase in body mass. Antral follicles grow slowly during the period of high circulating androstenedione concentration from October to December. There was a sharp decline in androstenedione concentration during January. Simultaneously with the decline in androstenedione concentration, a sharp increase in size of the antral follicle and circulating estradiol concentration was noticed. Soon thereafter, one of the follicles ruptures, followed by fertilization and the commencement of the first pregnancy. During the second pregnancy, antral follicles first appeared during late pregnancy in March in the contralateral ovary that lacked the corpus luteum. One follicle developed rapidly and quickly attained a preovulatory stage in April. This is reflected in a sharp increase in estradiol concentration during this period. Ovulation was observed immediately following the first pregnancy in May. During this period, androstenedione concentration remained low. The results of the present study suggest that high androstenedione concentration during October to December (winter dormancy) may be responsible for slow follicular development and delays ovulation in T. longimanus. It is further hypothesized that the geographical variation in reproductive pattern of T. longimanus could be due to variation in the duration of fat storage and associated changes in the androstenedione concentration. PMID- 11857473 TI - Effect of antiandrogen cyproterone acetate on the development of the antler cycle in Southern pudu (Pudu puda). AB - The antler cycle of pudu is similar to other cervids, but unlike most boreal deer, male Southern pudu (Pudu puda) exhibits two seasonal peaks of LH and testosterone. In that respect, pudu is similar to roe deer. Whereas the antler cycle in some deer species, such as roe deer or white-tailed deer, is very sensitive to variation of testosterone, in other cervids, such as fallow deer or reindeer, a blockade of androgens with cyproterone acetate (CA) has little or no effect on the timing of the antler casting. In order to test the sensitivity of pudu antlers to variations of androgens, CA (administered 2x weekly at 50 mg/buck) was injected intramuscularly for 3 weeks in 5 adult male pudu, starting February 19 (late summer). Four other males of similar age served as controls. The experiment was performed at the University of Concepcion, Chile, latitude 36.6 degreeS. Blood samples were taken once a week between January 19 and April 3. In CA-treated bucks, the antlers were cast approximately 3 weeks after the initiation of CA treatment and a new antler growth began almost immediately. The antlers reached about 5 cm in length, before ceasing to grow at the end of April, when they became mineralized and were subsequently polished. CA had no effect on the already declining levels of LH. Plasma levels of testosterone in controls increased from February 15, whereas in CA-treated bucks remained depresses until March 21. It is concluded that similarly to white-tailed deer, the antler cycle of Southern pudu is very sensitive to manipulation of androgen levels. PMID- 11857474 TI - Isolation of the peri-acrosomal plasma membrane protein D40 enriched fraction from guinea pig spermatozoa using a monoclonal antibody. AB - Membrane fragments were obtained from guinea pig spermatozoa by mechanical shearing. A membrane-enriched fraction was separated from other cellular debris, mainly sperm nuclei and tails, by centrifugation on 20% Ficoll 70 solution. Peri acrosomal plasma membrane protein, D40, enriched fraction was separated from this membrane preparation using a mouse monoclonal antibody to D40 attached to magnetic beads. Enrichment of D40 antigen in this fraction was demonstrated by western blotting. The method provides a preparative route to a membrane, the constituents of which play an important role in sperm recognition of the zona pellucida and the acrosome reaction. Some constituents of the peri-acrosomal plasma membrane over the equatorial segment of the acrosome may also play a role in sperm docking with the oocyte plasma membrane and fusion of the two cells. PMID- 11857475 TI - Membrane currents in the oocyte of the toad Bufo arenarum. AB - The amphibian oocyte cell model is widely used for heterologous expression of ionic channels and receptors. Little is known, however, about the physiology of oocyte cell models other than Xenopus laevis. In this study, the two-electrode voltage clamp technique was used to assess the most common electrical patterns of oocytes of the South American toad Bufo arenarum. Basal membrane resistance, resting potential, and ionic currents were determined in this cell model. The oocyte transmembrane resistance was 0.35 M(Omega), and the resting potential in normal saline was about -33 mV with a range between -20 mV and -50 mV. This is, to our knowledge, the first attempt to begin an understanding of the ion transport mechanisms of Bufo arenarum oocytes. This cell model may provide a viable alternative to the expression of ion channels, in particular those endogenously observed in Xenopus laevis oocytes. PMID- 11857476 TI - Deduced primary structure of vitellogenin in the giant freshwater prawn, Macrobrachium rosenbergii, and yolk processing during ovarian maturation. AB - A cDNA encoding vitellogenin (Vg) in the giant freshwater prawn, Macrobrachium rosenbergii, was cloned based on the cDNA sequence of vitellin (Vn) fragments A-N and B-42 determined previously, and its amino acid sequence deduced. The open reading frame (ORF) encoded 2,537 amino acid residues and its deduced amino acid sequence possessed three consensus cleavage sites, R-X-R-R, similar to those reported in Vgs of insects. The deduced primary structure of Vg in M. rosenbergii was seen to be similar to that of Penaeus japonicus, especially in the N-terminal region. It is therefore likely that Vgs in crustacean species including prawns and other related decapods exhibit a similar structural pattern. Based on the deduced primary structure of Vg and analysis of the various Vg and Vn subunits found in the hemolymph and ovary during ovarian maturation, we demonstrated the post-translational processing of Vg in M. rosenbergii. This is the first time that Vg processing has been clearly demonstrated in a crustacean species. Vg, after being synthesized in the hepatopancreas, is considered to be cleaved by a subtilisin-like endoprotease to form two subunits, A and proB, which are then released into the hemolymph. In the hemolymph, proB is possibly cleaved by a processing enzyme of unknown identity to give rise to subunits B and C/D. The three processed subunits A, B, and C/D are sequestered by the ovary to give rise to three yolk proteins, Macr-VnA, VnB, and VnC/D. PMID- 11857477 TI - Expression of SR-BI (Scavenger Receptor Class B Type I) in turtle (Chrysemys picta) tissues and other nonmammalian vertebrates. AB - In this study, the tissue distribution of the expression of an HDL-receptor (SR BI; Scavenger Receptor Class B Type I) was investigated in the turtle using an antiserum to murine SR-BI. Several turtle tissues including liver, heart, small intestine, kidney, oviduct, ovary, and testis were shown to express an 82 kDa membrane protein. In addition, SR-BI expression in livers of other nonmammalian species such as the chicken, frog, goldfish, shark, and skate is also reported. Ovarian SR-BI expression varies seasonally in the turtle as do plasma levels of apoA-I and cholesterol ester. It is possible that changing levels of SR-BI, the receptor for apoA-I, is physiologically relevant to the demands of the turtle ovarian cycle and cholesterol distribution. PMID- 11857478 TI - Retinoid antagonists inhibit normal patterning during limb regeneration in the axolotl, Ambystoma mexicanum. AB - Retinoic acid (RA) has been detected in the regenerating limb of the axolotl, and exogenous RA can proximalize, posteriorize, and ventralize blastemal cells. Thus, RA may be an endogenous regulatory factor during limb regeneration. We have investigated whether endogenous retinoids are essential for patterning during axolotl (Ambystoma mexicanum) limb regeneration by using retinoid antagonists that bind to specific RAR (retinoic acid receptor) or RXR (retinoid X receptor) retinoid receptor subtypes. Retinoid antagonists (Ro41-5253, Ro61-8431, LE135, and LE540) were administered to regenerating limbs using implanted silastin blocks loaded with each antagonist. The skeletal pattern of regenerated limbs treated with Ro41-5253 or Ro61-8431 differed only slightly from control limbs. Treatment with LE135 inhibited limb regeneration, while treatment with LE540 allowed relatively normal limb regeneration. When LE135 and LE540 were implanted together, regeneration was not completely inhibited and a hand-like process regenerated. These results demonstrate that interfering with retinoid receptors can modify pattern in the regenerating limb indicating that endogenous retinoids are important during patterning of the regenerating limb. PMID- 11857479 TI - Intensity-dependent phase-adjustments in the locomotor activity rhythm of the nocturnal field mouse Mus booduga. AB - The locomotor activity rhythm of the nocturnal field mouse Mus booduga was monitored under constant darkness (DD) and free-running periods (tau) were estimated. Following a free-run of about 15 days in DD, the animals were exposed to periodic light pulses (LPs) of various intensities (1 lux, 10 lux, 50 lux, 100 lux, and 1,000 lux) and 15 minutes duration for 65 days at intervals of 24 hours to investigate the influence of intensity of light on the phase-angle-difference (psi) between the onset of locomotor activity and the time of LP administration. The experimentally observed values of psi and tau for a LP of 1,000 lux intensity used for 15 minutes every 24 hr, showed a sigmoid shaped relationship with tau. This relationship was similar to that predicted based on the nonparametric model of entrainment, which uses the tau and the LP phase response curve (PRC) constructed using LP of similar duration and intensity. The functional nature of the relationship between psi and tau was not found to change significantly with increasing intensities of LP used to entrain the locomotor activity rhythm. However, psi was significantly modulated by the intensity of LP. These results suggest that the periodic sensitivity of the circadian pacemaker underlying the locomotor activity rhythm in the nocturnal field mouse M. booduga to LPs plays an important role in maintaining a characteristic psi with the zeitgeber and the psi changes in a light intensity-dependent manner. PMID- 11857480 TI - Energy expenditure and testosterone in free-living male yellow-pine chipmunks. AB - The onset of mating in yellow-pine chipmunks (Tamias amoenus) follows emergence from a prolonged period of energy conservation during hibernation. Energy expenditures are greatly accelerated to meet the demands of the reproductive season. When emerging from hibernation, typical male chipmunks (breeders) have enlarged testes and a high level of plasma testosterone (T). However, certain males that do not participate in reproduction (nonbreeders) maintain small testes and low plasma T levels and emerge several weeks later than the breeders. The timing of the terminal arousal from hibernation and onset of mating are associated with increased plasma T levels. Experimental elevation of T levels in T. amoenus outside the mating season has been associated with a decrease in body mass, further suggesting an effect of T on energy balance. To test this hypothesis, we measured daily energy expenditure (DEE) in free-living, nonbreeding male chipmunks in the presence and absence of a T-implant. We also measured DEE in breeding males when endogenous T levels were high. DEE of the nonbreeders was not affected by our manipulation of plasma T, and the DEE of breeding males did not differ from that of nonbreeders. We conclude that energy expenditure on a daily basis in male yellow-pine chipmunks is not influenced by levels of T. However, on a seasonal basis, the earlier emergence from hibernation by breeding males, which appears to be influenced by T, represents an overall seasonal energy expenditure that exceeds that of nonbreeding males. PMID- 11857481 TI - Effects of chemical anoxia on adrenergic responses of goldfish hepatocytes and the contribution of alpha- and beta-adrenoceptors. AB - Adrenergic responses during normoxia and chemical anoxia were investigated in anoxia-tolerant hepatocytes from the goldfish, Carassius auratus. Epinephrine stimulated glucose release was unaltered after 1 hr of chemical anoxia, the concentration of epinephrine required for half maximal stimulation of glucose release (K0.5(GLU)) ranging from 0.62 x 10(-8) to 2.05 x 10(-8) M. Similarly, the maximum rate of glucose release caused by hormonal stimulation was not affected by chemical anoxia. In anoxic goldfish hepatocytes [Ca2+](i) remained constant in nonstimulated cells but could be elevated by addition of epinephrine. The magnitude of this [Ca(2+)](i)-increase was dependent on the concentration of the catecholamine and this dependency was similar under normoxia (K0.5(Ca2+) = 1.17 x 10(-8) M) and chemical anoxia (K0.5(Ca2+) = 1.15 x 10(-8) M), as was the percentage of cells responding (77%) and displaying oscillatory [Ca2+]i response patterns (60%) after epinephrine addition, although the frequency of [Ca2+]i oscillations was significantly lower in anoxic cells. To analyze a possible shift in the importance of alpha- and beta-adrenoceptors during chemical anoxia, the effect of phentolamine and propranolol, alpha- and beta-adrenergic antagonists respectively, on epinephrine-stimulated glucose release was studied. Application of the alpha-antagonist caused a dose-dependent reduction of glucose-release which was similar under both conditions, whereas the sensitivity to the beta antagonist was lowered after chemical anoxia. Taken together these results provide evidence that during chemical anoxia goldfish hepatocytes remain responsive to adrenergic stimulation and that there is a partial shift regarding the contribution of alpha- and beta-adrenergic pathways to the induction of cellular glucose release stimulated by epinephrine. PMID- 11857482 TI - Modulation of goldfish testicular testosterone production in vitro by tumor necrosis factor alpha, interleukin-1beta, and macrophage conditioned media. AB - The relevance of immune-endocrine interactions to the regulation of testicular steroidogenesis in teleosts is virtually unexplored. The objectives of the present study were: 1) to investigate the effects of murine cytokines, tumor necrosis factor-alpha (TNFalpha) and interleukin-1beta (IL-1beta), and trout (Oncorhynchus mykiss) macrophage conditioned media (MCM) on testosterone (T) production by goldfish (Carassius auratus) testis pieces in vitro; and 2) to identify the site(s) of the inhibitory action of TNFalpha on hCG-stimulated T formation. TNFalpha (0-100 ng/ml) affected basal T production differentially depending on the gonadosomatic index (GSI) value of the fish. TNFalpha stimulated basal T of fish with a relatively low GSI (average 1.99), but inhibited T production by testis of fish with a higher GSI (average 5.14). The remaining studies used fish with only high GSI values. IL-1beta (0-10 ng/ml) inhibited basal T production, while MCM (0-25% v/v) had no effect. The cytokines significantly inhibited hCG-stimulated T production at all doses tested, whereas MCM was inhibitory only at the lower doses of 2.5-5% v/v. TNFalpha did not affect basal or hCG-stimulated cAMP levels, but did inhibit forskolin (0.5 microM; adenylate cyclase activator) and 8-bromo-cAMP (0.15 mM; cAMP analog) stimulated T levels. The inhibitory actions of TNFalpha on T production were greatly reduced by treatment of testis with 25-hydroxycholesterol (1 and 10 microg/ml), pregnenolone (50 and 100 ng/ml), and 17 alpha-hydroxypregesterone (50 ng/ml). TNFalpha caused a moderate decrease in pregnenolone (100 ng/ml)-stimulated T production. Together, these data demonstrate that regulatory actions of TNFalpha may occur at multiple sites within the steroid biosynthetic pathway, but the major effect appears to be related to cholesterol availability in the mitochondria. In conclusion, the results of this study implicate macrophage derived factors in the regulation of teleost testicular androgen biosynthesis. PMID- 11857483 TI - Costs of anorexia during pregnancy in a viviparous snake (Vipera aspis). AB - Spontaneous anorexia has been documented in various animal species and is usually associated with activities competing with food intake. In natural conditions, most female aspic vipers (Vipera aspis) stop feeding during the two months of pregnancy. We carried out a simple experiment on 40 pregnant females to determine whether anorexia was obligatory or facultative, and to investigate the energetic consequence of fasting on post-partum body condition and litter traits. Three diet treatments were applied during gestation: no food, one feeding occasion, and two feeding occasions. Twelve nonpregnant, unfed females were used as a control group. Most gravid females accepted captive mice during gestation, suggesting that anorexia reported in the field was a side effect of the tremendous changes in activity pattern associated with pregnancy. Mass loss was high for unfed reproductive females, indicating high energy expenditure associated with embryo maintenance. Prey consumption allowed compensation for metabolic expenditure and enhanced post-partum female body condition, but had no effects on litter characteristics. The magnitude of the metabolic expenditure during gestation appeared to be independent of fecundity. PMID- 11857484 TI - Temperature effects and genotype-temperature interactions on sex determination in the European sea bass (Dicentrarchus labrax L.). AB - The effect of temperature on sex-ratios in 27 families of sea bass reared in the same tank from the fertilization stage onward was investigated. An excess of males (68%) was found in the groups that were reared at high temperature (mean +/ standard deviation: 20+/-1 degrees C) until they reached the mean size of 8.1 cm (Standard Length, 149 days post-fertilization [p.f.]). Masculinization was higher (89% of males) in the groups maintained at low temperature (13 degrees C), from fertilization to a mean length of 6.5 cm (346 days p.f.). Shifts from high to low temperature at 8.1cm and from low to high temperature at 6.5 cm had no consequence on the sex-ratio. The percentage of males showing intratesticular oocytes was higher at low temperature (63%) than at high temperature (36%), suggesting that these males may be sensitive fish that have been masculinized by environmental factors. Fish sampled in the groups reared at high (2,200 fish) and low (500 fish) temperature were genotyped on three microsatellite loci. This allowed them to be assigned to the breeders used in the crossing design, thus permitting an analysis of parental influence on sex-ratios. In groups reared at high temperature, both parents had a significant additive effect on the percentage of females, and the interaction between sire and dam was not significant. Genotype temperature interactions were also detected and their existence suggests the interesting possibility of selecting nonsensitive genotypes in breeding programs. PMID- 11857486 TI - HLA-G and its KIR ligands in cancer--another enigma yet to be solved? AB - Alterations of antigen-presenting human leukocyte antigen (HLA) molecules often occur in cancer and represent one of the mechanisms by which tumours evade host immunosurveillance. One such molecule, HLA-G, was recently implicated in the same context. HLA-G is thought to mediate the tolerance of the semi-allogeneic fetus during pregnancy, by inhibiting maternal immune response through interaction with different HLA-G-recognizing killer-cell inhibitory receptors (KIRs). A report in this issue of the journal demonstrates for the first time the expression of HLA-G and its KIR ligand, ILT-2 in human breast cancer. Apart from the up-regulation on tumour cells, the authors describe HLA-G induction on the macrophages and T cells infiltrating the breast cancer lesions. Moreover, a subset of infiltrating cells also expressed HLA-G-recognizing KIR, ILT-2. This editorial discusses the induction of HLA-G and related KIR molecules as another mechanism for tumours to escape immune recognition. PMID- 11857487 TI - The role of tumour-associated macrophages in tumour progression: implications for new anticancer therapies. AB - The role of macrophages in tumour growth and development is complex and multifaceted. Whilst there is limited evidence that tumour-associated macrophages (TAMs) can be directly tumouricidal and stimulate the anti-tumour activity of T cells, there is now contrasting evidence that tumour cells are able to block or evade the activity of TAMs at the tumour site. In some cases, tumour-derived molecules even redirect TAM activities to promote tumour survival and growth. Indeed, evidence has emerged for a symbiotic relationship between tumour cells and TAMs, in which tumour cells attract TAMs and sustain their survival, with TAMs then responding to micro-environmental factors in tumours such as hypoxia (low oxygen tension) by producing important mitogens as well as various growth factors and enzymes that stimulate tumour angiogenesis. This review presents evidence for the number and/or distribution of TAMs being linked to prognosis in different types of human malignancy. It also outlines the range of pro- and anti tumour functions performed by TAMs, and the novel therapies recently devised using TAMs to stimulate host immune responses or deliver therapeutic gene constructs to solid tumours. PMID- 11857488 TI - Specific activation of the non-classical class I histocompatibility HLA-G antigen and expression of the ILT2 inhibitory receptor in human breast cancer. AB - The HLA-G molecule is a non-classical HLA class I antigen selectively expressed by trophoblastic cells that invade the maternal decidua during human pregnancy. HLA-G is believed to contribute to tolerance of the semi-allogeneic fetus by inhibiting maternal immune responses. Similarly, HLA-G expression in tumour cells may favour their escape from host immune surveillance. This study investigated HLA-G expression in human mammary tumours. Immunohistochemical analysis of cryo preserved and paraffin-embedded breast tissue biopsies, using two HLA-G-specific antibodies, revealed that unlike non-cancerous breast tissue in the vicinity of the tumour, 14 out of 36 breast cancer lesions selectively expressed HLA-G. HLA-G expression was significantly more frequent in lesions that were highly infiltrated by host immune cells, thus correlating HLA-G activation with inflammation. Further histological and double-staining immunofluorescence analysis attributed HLA-G expression mainly to tumour epithelial cells and to subsets of infiltrating CD68+ and CD8+ cells. RT-PCR analysis suggested that HLA G was activated at the transcriptional level in breast tumours. The presence of ILT2 (Ig-like transcript 2) killing inhibitory receptors known to interact with HLA-G was also demonstrated in host immune cells that infiltrate breast cancer lesions. These results indicate that HLA-G is up-regulated at high frequencies in human breast cancer, where it may impair efficient anti-tumour immunity. PMID- 11857490 TI - Genetic pathways in the evolution of breast ductal carcinoma in situ. AB - The patterns of allelic loss in 28 cases of pure ductal carcinoma in situ (DCIS) and 25 cases of DCIS associated with invasive ductal carcinoma (IDC) were compared, in order to define whether pure DCIS represented an earlier stage than DCIS associated with IDC in the progression of breast carcinoma. To this aim, the polymerase chain reaction (PCR) was performed on microdissected normal and neoplastic breast tissue, formalin-fixed and paraffin-embedded. Fifteen microsatellite markers were examined, on chromosomes 1p, 3p, 7q, 11q, 12p, 13q, 16q and 17q, mostly focused on regions altered in breast cancer. Loss of heterozygosity (LOH) was greater in pure DCIS than in the DCIS component associated with IDC for 11 out of 15 markers. The difference was statistically significant for D13S260 and D17S800 (p=0.008 and p=0.01, respectively). DCIS associated with IDC showed a lesser degree of alteration than the synchronous IDC component for ten out of 15 markers. In contrast, LOH at D11S1816 and D16S318 was lower in pure DCIS than in DCIS associated with IDC and even greater in the IDC component. These results confirm that DCIS is a possible but not an obligate precursor of invasive breast cancer and suggest that pure DCIS and DCIS associated with IDC may be genetically distinct. The evolution from DCIS to IDC may follow multiple pathways and not a linear model. PMID- 11857489 TI - Mitotic counts provide additional prognostic information in grade II mammary carcinoma. AB - The ability to predict how long a patient diagnosed with breast cancer is likely to survive is still imprecise, despite numerous studies which have identified potential prognostic markers. The "established" markers such as nodal status, tumour size, and histological grade have been used for many years and certainly provide some degree of accuracy upon which treatment can be based. However, women with similar prognostic features can vary significantly in their outcome and very few of the newly identified markers provide information that is sufficiently useful to warrant the time and expense spent on their evaluation. In a cohort of 145 women, an assessment has been made of whether knowledge of the proliferative activity of grade II infiltrating ductal breast carcinomas can improve the accuracy of predicting clinical outcome for individual patients. Use of the mitotic count (MC), which was assessed as part of the grading system, enabled patients to be stratified into "good" and "bad" prognostic groups. The measurement of S-phase fraction using flow cytometry gave a similar result, but has the disadvantage that the technique requires specialized equipment. The evaluation of Ki-67 expression using immunohistochemistry was of no additional prognostic value in this defined group. It is proposed that MC, used once to establish grade, could be used again amongst the grade II tumours to improve the accuracy of prognosis and thus influence treatment strategies with minimal additional effort or expense. PMID- 11857491 TI - Loss of heterozygosity and allelic imbalance in apocrine metaplasia of the breast: microdissection microsatellite analysis. AB - Loss of heterozygosity (LOH) and allelic imbalance (AI) at loci reported to show allele loss and/or imbalance in preinvasive and invasive breast cancer were examined in 41 cases of apocrine metaplasia (APM) of the breast using a microdissection technique, polymorphic microsatellite markers, and the polymerase chain reaction (PCR). Occasional examples of LOH and/or AI were identified in 2/28 (7.1%) informative cases at 1p (MYCL1), 2/14 (14.3%) at 11q (INT2), 1/15 (6.7%) at 13q (D13S267), 3/22 (13.6%) at 16q (D16S539), 2/23 (8.7%) at 17p (TP53), and 1/11 (9.1%) at 17p (D17S513) and 3/16 (18.8%) at 17q (D17S250). The finding of LOH/AI in cases of APM indicates that a subset of APM appears clonal, but the significance of allelic loss or imbalance in the pathogenesis of APM or its possible subsequent progression to carcinoma is not yet clear and requires further investigation. Clinical follow-up of these particular cases of APM showing LOH/AI would be of further value. PMID- 11857492 TI - Expression of S100A4 protein is associated with metastasis and reduced survival in human bladder cancer. AB - The calcium-binding protein S100A4 induces the metastatic phenotype in rodent models of breast cancer and its expression correlates strongly with reduced survival in human breast cancer. The expression of S100A4 in normal bladders and 101 bladder tumours has been studied using immunocytochemistry. Moderate or strong expression of S100A4 was found in 28% of the tumours, whilst the remaining tumours and normal urothelium either failed to stain or showed weak staining. S100A4 staining was more frequently observed in invasive bladder tumours than in non-invasive tumours (p<0.05). In invasive tumours, S100A4 staining was usually strongest in invasive regions and single infiltrating cells. Statistically significant associations were found between S100A4 expression and metastasis (p=0.0003) and reduced survival (p<0.0001). It is concluded that S100A4 expression may play an important role in bladder cancer and may identify a subgroup of patients at increased risk of metastasis who should be considered for adjuvant systemic therapy. PMID- 11857493 TI - Multiple molecular alterations of FHIT in betel-associated oral carcinoma. AB - To determine the alterations of the FHIT (fragile histidine triad) gene in oral squamous cell carcinoma (OSCC), this study examined mutation, promoter methylation, mRNA transcription, and protein expression of FHIT in OSCC associated mostly with the use of betel and/or tobacco. Analyses of the coding exons (exons 5-9) identified a deletion of one base in intron 4 in one tumour and a deletion of exon 7 in two tumours. Using bisulphite genomic sequencing, 28% of the informative subjects exhibited promoter methylation. An aberrant FHIT transcript spanning from exon 3 to exon 10, which was verified by RT-PCR analysis, was identified in 36% of the OSCC subjects, 50% of the oral pre invasive lesions, and 5% of the non-cancerous match tissue. An abnormal immunohistochemical level of Fhit was detected in 41% of OSCC subjects. A statistically significant association was found between aberrant transcription of the FHIT gene and an abnormal level of Fhit immunoreactivity. The results indicated that alteration of FHIT is a frequent occurrence in OSCC and thus suggests that the aberrance in FHIT transcription could be an early event of oral carcinogenesis. PMID- 11857494 TI - Apoptosis in B-cell lymphomas and reactive lymphoid tissues always involves activation of caspase 3 as determined by a new in situ detection method. AB - In vitro studies indicate that in lymphomas, execution of apoptosis involves activation of effector caspases. To investigate activation of effector caspases in vivo in biopsy specimens of lymphomas, a new assay was developed using antibodies against active caspase 3 and p89, a protein fragment generated by caspase-specific cleavage of poly-ADP ribose polymerase (PARP). Using this assay, it was found that in B-cell lymphomas, levels of active caspase 3/p89-positive cells correlate strongly with morphologically recognizable apoptotic cells. The number of active caspase 3/p89-positive cells was low in follicular lymphomas and usually high in diffuse large cell lymphomas. Highest numbers were found in Burkitt lymphomas and in two biopsies of diffuse large B-cell lymphomas (DLCLs) obtained several days after initiation of therapy. It is concluded that apoptosis in reactive lymphoid tissues and in B-cell lymphomas always involves activation of effector caspase 3 and cleavage of one of the major effector caspase substrates, PARP-1. Moreover, levels of effector caspase activation are constantly low in low-grade follicular lymphomas and vary considerably in DLCL and Burkitt lymphoma. PMID- 11857496 TI - Lumican expression in alpha cells of islets in pancreas and pancreatic cancer cells. AB - Lumican is a member of a small leucine-rich proteoglycan family, members of which play an important role in cell migration and proliferation during embryonic development, tissue repair, and tumour growth. Lumican is reported to be overexpressed during the wound healing process in the cornea and in human breast cancer tissues, but its expression and localization in normal pancreas and pancreatic cancer tissues are not known. The present study aimed to clarify the expression of lumican protein and its mRNA in human pancreatic cancer cell lines and their localization in normal and cancerous human pancreatic tissues. Reverse transcription-polymerase chain reaction (RT-PCR) and western blot analysis revealed lumican mRNA and its protein expression in PK-8 and MIA-PaCa-2 human pancreatic cancer cells. The tumour lumican had non- or poorly sulphated polylactosamine side-chains rather than highly sulphated keratan sulphate chains. Immunoreactivity of the lumican protein was localized in alpha cells of islets and stromal tissues of a normal pancreas. In pancreatic cancer tissues, the lumican protein was strongly localized in cancer cells, and in acinar and islet cells in chronic pancreatitis-like lesions adjacent to the cancer cells. It was also localized in fibroblasts and collagen fibres close to cancer cells. Lumican mRNA was expressed in cancer cells, in acinar and islet cells in chronic pancreatitis-like lesions, and in stromal fibroblasts in the pancreatic cancer tissues. This is the first report that lumican is synthesized in endocrine and cancer cells. Lumican protein may play a role in the maintenance of islet cell function in normal pancreas and the lumican protein synthesized by cancer cells, acinar and islet cells, and stromal fibroblasts may play a role in the growth of human pancreatic cancer cells. PMID- 11857495 TI - Overexpression of peroxiredoxins I, II, III, V, and VI in malignant mesothelioma. AB - Peroxiredoxins (Prxs) are a recently characterized group of thiol-containing proteins with efficient antioxidant capacity, capable of consuming hydrogen peroxide in living cells. Altogether six distinct Prxs have been characterized in mammalian tissues. Their expression was investigated in histological samples of mesothelioma and in cell lines established from the tumours of mesothelioma patients. Four cases with histopathologically healthy pleura from non-smokers were used as controls. Healthy pleural mesothelium was negative or very weakly positive for all Prxs. In mesothelioma, the most prominent reactivity was observed with Prxs I, II, V, and VI. Prx I was highly or moderately expressed in 25/36 cases, the corresponding figures for Prxs II-VI being 27/36 (Prx II), 13/36 (Prx III), 2/36 (Prx IV), 24/36 (Prx V), and 30/36 (Prx VI). Positive staining was observed both in the cytosolic and the nuclear compartment, with the exception of Prx III, which showed no nuclear reactivity. The staining pattern of Prxs III and V was granular. Immunoelectron microscopic localization of Prxs was in accordance with the immunohistochemical findings, showing diffuse cytoplasmic localization for Prxs I, II, IV, and VI and distinct mitochondrial labelling for Prxs III and V. There was no significant association between the extent of staining and different Prxs. It appeared that Prxs may not have prognostic significance, but being prominently expressed in most mesotheliomas these proteins, at least in theory, may play a role in the primary drug resistance of this disease. PMID- 11857497 TI - Methylation-specific PCR for detection of neoplastic DNA in biopsy washings. AB - Epigenetic DNA alterations such as promoter hypermethylation of glutathione S transferase P1 (GSTP1) in prostatic adenocarcinoma frequently constitute tumour biomarkers. Neoplastic transformation was identified in washings of prostate biopsies by GSTP1 promoter hypermethylation, using methylation-specific PCR (MSP). Twenty-six patients undergoing sextant transrectal prostate biopsies for clinically suspected prostate cancer were enrolled. All ten patients diagnosed with adenocarcinoma (100%) and four of six patients with prostatic intraepithelial neoplasia (67%), but none of ten patients with benign hyperplasia (0%), exhibited GSTP1 promoter hypermethylation in at least one out of six biopsy washings. Since this approach is transferable to various cancer entities, a sensitive and specific DNA-based analysis of biopsy material seems generally feasible without impeding routine histopathological examination. PMID- 11857498 TI - Expression of Bcl-x, Bcl-2, Bax, and Bak in endarterectomy and atherectomy specimens. AB - The regulation of apoptosis in atherosclerosis is not completely defined. The aim of this study was to determine the expression of Bcl-2, Bcl-x, Bax, and Bak in relation to apoptosis in advanced atherosclerotic lesions. In atherectomy (15), endarterectomy (10), and control non-atherosclerotic segments of renal (2) and of coronary and carotid (5) arteries, the extent of apoptosis was determined using TdT dUTP nick end labelling (TUNEL) and nuclear morphology (karyorrhexis/pyknosis) and expression of apoptosis regulators by immunohistochemistry and western blot analysis on paraffin-embedded material. In all specimens, the atherosclerotic involvement was advanced: grade V (n=18) and grade VI (n=7). The apoptotic index was high (mean 30%) in advanced lesions compared with controls (<2%) and smooth muscle cells (SMCs) were the predominant cell type undergoing apoptosis. In all TUNEL-positive apoptotic cells, Bax and Bak were present, while Bcl-x was absent. Bcl-2 was absent in a majority of these cells, but occasional TUNEL-positive cells expressed Bcl-2. In non-apoptotic cells, Bcl-x was present and western blot detected only the long isoform, Bcl-xL, from the plaques. In conclusion, increased Bax and Bak coupled with lack/paucity of Bcl-2 and Bcl-xL are associated with SMC apoptosis in advanced lesions. Bcl-xL in non-apoptotic cells appears to contribute to prolonged cell survival. PMID- 11857499 TI - Macrophages expressing the scavenger receptor CD163: a link between immune alterations of the gut and synovial inflammation in spondyloarthropathy. AB - The objective of this study was to investigate CD163+ macrophages in the synovial membrane of patients with spondyloarthropathy (SpA). Immunohistochemistry was performed on synovium of 17 SpA and 18 rheumatoid arthritis (RA) patients, on colonic biopsies of 16 SpA patients and ten healthy controls, and on paired synovial biopsies of eight SpA patients, before and after anti-TNFalpha therapy. Phenotype and cytokine production were analysed by flow cytometry. CD163+ macrophages were increased in the synovial lining and sublining in SpA versus RA, as well as in colonic lamina propria in SpA versus controls. The number of CD163+ macrophages in the synovial sublining correlated with C-reactive protein levels and erythrocyte sedimentation rate. Paralleling the increase of CD163, HLA-DR was increased in the synovial lining and sublining of SpA. In contrast, the co stimulatory molecules CD80 and CD86 and the dendritic cell markers CD1a and CD83 were scarce in SpA synovium. Flow cytometry indicated that CD163+ macrophages expressed high levels of HLA-DR and could produce in vitro tumour necrosis factor alpha (TNFalpha) but not interleukin-10 (IL-10). Finally, anti-TNFalpha therapy in vivo induced a decrease of CD163+ macrophages in the synovial lining and sublining. In conclusion, macrophages expressing the scavenger receptor CD163 are increased in synovium and in colonic mucosa in SpA, highlighting the relationship between joint and gut in this disease. The correlation with inflammatory parameters, the expression of HLA-DR, the production of TNFalpha but not IL-10, and the reduction by anti-TNFalpha therapy support a role for CD163+ macrophages in the synovial inflammation in SpA. PMID- 11857500 TI - Telepathology in a routine clinical environment: implementation and accuracy of diagnosis by robotic microscopy in a one-stop breast clinic. AB - The concept of using telepathology as a way of increasing the efficiency of pathologists is not new. There have been many studies attempting to evaluate the role of telepathology diagnosis, predominantly using transmission of still digital images. This study evaluates the potential value of remote diagnosis using robotic microscopy in the setting of a one-stop breast clinic. A Zeiss Axiopath telepathology system was used. The aim was to identify deficiencies in software and the minimum specifications for the computer hardware and network capability needed for reproducible pathological diagnosis with a view to developing a system that can preclude the need for an on-site pathologist. Forty seven fine needle aspiration smears were diagnosed simultaneously by a pathologist in the breast clinic and by a different pathologist operating a robotic microscope situated in the clinic in a different wing of the hospital from the pathology department. The diagnoses, the time taken for clinic and remote diagnosis, and difficulties in using the system were recorded. Absolute correlation was achieved in 80.9% of cases. There was one false-positive diagnosis of cancer and no false negatives. The mean time taken for diagnosis per case was 2.39 min in clinic and 10.81 min by remote control robotic microscopy. However, as the pathologist did not have to leave the department, remote diagnosis was overall more economical of pathologists' time. Image quality was generally found to be good and not an obstacle to diagnosis. There were operational and technical problems that made remote diagnosis tedious and lengthy. Although at present the system is not capable of replacing an on-site pathologist, the results were encouraging and suggest that remote control remote diagnosis has the potential to increase the efficiency of pathologists. PMID- 11857501 TI - The boundaries of cellular pathology: how pathologists see their clinical role. AB - There has been much recent public attention on the effects of the practices of cellular pathology on patients and their relatives. It is important to establish clearly pathologists' views about their professional relationships to clinicians and patients. A national postal questionnaire survey was performed to investigate how cellular pathologists perceive their role in clinical practice and whether there are important differences between different groups of pathologists. Responses were received from 773 pathologists, of whom 167 were trainees. On the basis of responses to 25 attitude statements, it appears that the profession shares a core belief that pathologists are clinicians rather than scientists and sees diagnosis as its main clinical responsibility. However, the role that each pathologist feels clear about differs significantly between pathologists, with a stronger consensus over function, responsibility, and professional image than on the clinical practice of pathology, job satisfaction, and the impact of technological change. Academic activities are a minority interest, although academic pathologists express better job satisfaction and more radical views of professional practice. Young trainees are more likely to work in teaching hospitals, to be female, to have trained outside the UK, and to have had experience in other specialties. They have, however, a more restricted view of the clinical role of pathology than accredited pathologists. Most cellular pathologists see themselves as clinicians with a special role in diagnosis, but how this is interpreted and the extent to which pathologists involve themselves in clinical decision-making is inconsistent. This professional dilemma should be addressed by research to obtain more systematic knowledge of the clinical impact of cellular pathology and especially the ill-defined links between diagnosis and clinical decision-making. PMID- 11857502 TI - Re. article entitled "Expression of CD31 by cells of extensive ductal in situ and invasive carcinomas of the breast". PMID- 11857504 TI - The lymphatic system in cancer. PMID- 11857505 TI - Priming the search for HOX mutations. PMID- 11857506 TI - Complete mutation analysis panel of the 39 human HOX genes. AB - BACKGROUND: The HOX gene family consists of highly conserved transcription factors that specify the identity of the body segments along the anteroposterior axis of the embryo. Because the phenotypes of mice with targeted disruptions of Hox genes resemble some patterns of human malformations, mutations in HOX genes have been expected to be associated with a significant number of human malformations. Thus far, however, mutations have been documented in only three of the 39 human HOX genes (HOXD13, HOXA13, and HOXA11) partly because current knowledge on the complete coding sequence and genome structure is limited to only 20 of the 39 human HOX genes. METHODS: Taking advantage of the human and mouse draft genome sequences, we attempted to characterize the remaining 19 human HOX genes by bioinformatic analysis including phylogenetic footprinting, the probabilistic prediction method, and comparison of genomic sequences with the complete set of the human anonymous cDNA sequences. RESULTS: We were able to determine the full coding sequences of 19 HOX genes and their genome structure and successfully designed a complete set of PCR primers to amplify the entire coding region of each of the 39 HOX genes from genomic DNA. CONCLUSIONS: Our results indicate the usefulness of bioinformatic analysis of the draft genome sequences for clinically oriented research projects. It is hoped that the mutation panel provided here will serve as a launchpad for a new discourse on the genetic basis of human malformations. PMID- 11857507 TI - Effects of folate supplementation on the risk of spontaneous and induced neural tube defects in Splotch mice. AB - BACKGROUND: Neural tube defects (NTDs) are among the most common human congenital malformations. Although clinical investigations have reported that periconceptional folic acid supplementation can reduce the occurrence of these defects, its mechanism remains unknown. Therefore, the murine mutant Splotch, which has a high incidence of spontaneous NTDs, along with the inbred strains SWV and LM/Bc, were used to investigate the relationship between folate and NTDs. METHODS: To investigate whether folates could reduce spontaneous NTDs, heterozygous Splotch dams (+/Sp) were treated with either folate or folinic acid throughout neurulation, gestational day (GD) 6.5 to 10.5. On GD 18.5 the dams were sacrificed and the fetuses examined for any neural tube defects. Subsequently, Sp/+ dams were treated with arsenic while receiving either a folate or folinic acid supplementation. Similar experiments were performed in the LM/Bc and SWV strains. RESULTS: Neither folate nor folinic acid supplements reduced the frequency of spontaneous NTDs in the embryos from Splotch heterozygote crosses. Arsenic increased the frequency of NTDs and embryonic death in the Splotch, LM/Bc and SWV litters and folinic acid failed to ameliorate the teratogenic effect of this metal. A folate supplement given to arsenic-treated dams proved to be maternally lethal in all three strains. CONCLUSIONS: Splotch embryos were not protected from either spontaneous or arsenic-induced NTDs by folinic or folic acid supplementation. Furthermore, folinic acid supplements did not reduce the incidence of arsenic-induced NTDs in either the LM/Bc or SWV litters. PMID- 11857508 TI - Depletion of Mab21l1 and Mab21l2 messages in mouse embryo arrests axial turning, and impairs notochord and neural tube differentiation. AB - BACKGROUND: The nematode mab-21 gene specifies sensory ray cell identity and was first isolated because of its mutant sensory ray defects. Vertebrate Mab21 orthologs have since been identified in mammals and amphibians. In this report, we characterized in detail two Mab21 orthologs in mouse, Mab21l1 and Mab21l2. METHODS: We examined the genomic organizations of Mab21 genes and used northern blot and in situ hybridizations to assay their temporal-spatial expression pattern. Their embryonic functions were revealed by specific attenuation of Mab21 messages with antisense oligos in cultured embryos. RESULTS: Mab21l1 and Mab21l2 have very similar protein make-up and gene structures. Both genes were expressed in overlapping domains of actively differentiating embryonic tissues. In addition, Mab21l1 had unique expression in the lens vesicles and genital tubercle whereas Mab21l2 was expressed in the retinal epithelium and umbilical cord. Mab21l1 and Mab21l2 depleted embryos had severe defects in notochord, neural tube, organogenesis, vasculogenesis, and axial turning. CONCLUSIONS: The findings demonstrate that both Mab21 genes are required in developing embryos for embryonic turning, formation of the notochord, neural tube, and other organ tissues. PMID- 11857509 TI - Design and development of an Internet registry for congenital heart defects. AB - BACKGROUND: Congenital Heart Defects (CHD) are conditions that encompass more than 50 diagnoses and are due to developmental abnormalities early in fetal life. The King Faisal Specialist Hospital and Research Centre in the Kingdom of Saudi Arabia treats approximately 100 new cases per month. We recently developed a new CHD Registry that captures, stores and processes our data via the Internet. METHODS: The Registry was developed using Hypertext Markup Language (HTML), Microsoft Active Server Pages and Microsoft Structured Query Language (SQL). RESULTS: Details of CHD cases are captured in a World Wide Web (WWW) Registry, permitting any browser-enabled PC or Mac to participate fully in all registry functions, including data-entry, viewing, editing, searching, reporting, validating, charting, and exporting data subsets to statistics packages. It includes "administrative" features and an active security system. The paper forms have been designed to reflect the "look and feel" of the Web pages. Automatic validation procedures are also included. CONCLUSIONS: Our Registry has been in operation for 3 years. It serves 10 PCs and contains more than 3,000 registered cases of CHD. It is the first CHD Registry to be fully functional on the Internet. It is also the first dedicated CHD registry, and the first to routinely report on the full spectrum of CHD diagnoses. The WWW offers several logistical advantages to disease registries, especially those that represent large regions. It also offers the possibility of sharing resources between registries, facilitating the aggregation and analysis of disease data on a world-wide scale. This is useful for rare diseases such as CHD (see http://rc.kfshrc.edu.sa/chdr/demo/). PMID- 11857510 TI - Sonographic assessment of blunt abdominal trauma: a 4-year prospective study. AB - PURPOSE: Emergency abdominal sonography has become a common modality worldwide in the evaluation of injuries caused by blunt trauma. The sensitivity of sonography in the detection of hemoperitoneum varies, and little is known about the accuracy of sonography in the detection of injuries to specific organs. The purpose of this study was to determine the overall accuracy of sonography in the detection of hemoperitoneum and solid-organ injury caused by blunt trauma. METHODS: From January 1995 to October 1998, 3,264 patients underwent emergency sonography at our institution to evaluate for free fluid and parenchymal abnormalities of specific organs caused by blunt trauma. All patients with intra-abdominal injuries (IAIs) were identified, and their sonographic findings were compared with their CT and operative findings, as well as their clinical outcomes. RESULTS: Three hundred ninety-six (12%) of the 3,264 patients had IAIs. Sonography detected free fluid presumed to represent hemoperitoneum in 288 patients (9%). The sonographic detection of free fluid alone had a 60% sensitivity, 98% specificity, 82% positive predictive value, and 95% negative predictive value for diagnosing IAI. The accuracy was 94%. Seventy patients (2%) had parenchymal abnormalities identified with sonography that corresponded to actual organ injuries. The sensitivity of the sonographic detection of free fluid and/or parenchymal abnormalities in diagnosing IAI was 67%. CONCLUSIONS: Emergency sonography to evaluate patients for injury caused by blunt trauma is highly accurate and specific. The sonographic detection of free fluid is only moderately sensitive for diagnosing IAI, but the combination of free fluid and/or a parenchymal abnormality is more sensitive. PMID- 11857511 TI - Two-dimensional and three-dimensional sonography of conjoined twins. AB - PURPOSE: The aim of this study was to evaluate and compare the diagnostic capabilities of 2-dimensional (2D) and 3-dimensional (3D) sonography for the study of conjoined twins. METHODS: Four pregnant women with an initial 2D sonographic diagnosis of conjoined twins were examined with color Doppler sonography, 3D multiplanar sonography, and orthogonal plane imaging and 3D surface rendering. RESULTS: All 4 cases of conjoined twins were initially diagnosed with either transvaginal or transabdominal 2D sonography. 3D sonography afforded more realistic views and demonstrated more clearly the linking areas and surface anomalies, but 2D and color Doppler sonography provided more definitive and specific information about shared organs. CONCLUSIONS: Although 2D sonography is the primary modality for diagnosing and evaluating conjoined twins, color Doppler and 3D sonography can sometimes provide additional information that assists in the clinical management of these twins. 3D sonography also provides images that are easier for parents to understand, which can help them with decision making. PMID- 11857512 TI - Borderline cystic tumors of the ovary: gray-scale and color Doppler sonographic findings. AB - PURPOSE: The aim of the study was to determine the value of gray-scale and color Doppler sonography in distinguishing borderline cystic tumors (BCTs) from benign cysts and malignant tumors of the ovary. METHODS: The gray-scale and color Doppler sonographic features of 383 ovarian lesions in 374 nonpregnant women were retrospectively studied. Sonography was performed transvaginally for all but 7 lesions, which were imaged suprapubically. All of the lesions were surgically resected via laparoscopy or laparotomy. RESULTS: The histopathologic diagnoses were 27 BCTs, 35 ovarian carcinomas, and 321 benign cysts. Sonography diagnosed 24 (89%) of 27 BCTs as malignant lesions. Patients with BCTs, were younger than those with ovarian cancer (p < 0.001). BCTs showed intracystic papillae in 17 cases (63%), diffuse internal echoes in 11 (41%), intracystic septa in 8 (30%), a heterogeneous echo pattern in 7 (26%), and a solid pattern in 4 (15%). BCTs showed blood flow in 24 cases (89%) and lower pulsatility and resistance indices (RI) compared with benign lesions (p < 0.001 for both). Multivariate analysis revealed intracystic papillae as the only independent predictor of BCTs (p < 0.001). CONCLUSIONS: When a cystic mass has papillae, this is the only abnormal finding detected by gray-scale transvaginal sonography, and color Doppler imaging shows low RI values within the mass, a BCT should be suspected. PMID- 11857513 TI - Sonographic examination of the oral phase of swallowing: bolus image enhancement. AB - PURPOSE: The purpose of this study was to evaluate the ability of 4 liquid boluses to enhance pixel brightness and the ease with which the boluses could be identified during the sonographic evaluation of oral swallowing in healthy young adults. METHODS: Ten healthy adult volunteers (5 men and 5 women), ranging in age from 21 to 31 years, underwent sonographic evaluation of the oral phase of swallowing while sitting in their usual feeding position. We compared the ability of the 4 following liquids to improve sonographic visualization of swallowing with that of water: a carbonated cola beverage, 5.0 ml of Thick-It in 120 ml of water, 2.5 ml of Thick-It in 120 ml of water, and 7.5 ml of confectioners' sugar in 120 ml of water. Water was used as a control. In each case, 5 ml of the liquid was introduced into the subject's oral cavity using a syringe, and the subject was instructed to swallow. Digitized still images and recorded video sequences of sonographic examinations of the swallowing were analyzed. The brightness of the bolus image on selected digitized video frames was measured digitally using Image Analyst software. Pixel brightness within selected regions of interest for each of the test liquids was statistically compared with that for water. Seven clinicians rated the visualization of each test liquid and water on paired sonographic videotape sequences. These ratings and the level of agreement between them were statistically tested. RESULTS: Only the carbonated cola beverage demonstrated statistically greater pixel brightness than that of water on digitized video frames (p = 0.01), whereas both cola (with a moderate inter-rater agreement, kappa = 0.50) and 5.0 ml Thick-It mixed with 120 ml of water (with a fair inter-rater agreement, kappa = 0.24) were significantly better visualized on sonographic video sequences. CONCLUSIONS: The digital still-frame analysis confirmed the clinicians' ratings of bolus visualization on real-time sonography, but dynamic sonography is more important than still frames in assessing sonographic swallow media because the dynamic images more closely parallel what is seen in clinical practice. Future investigations of sonographic contrast agents for use in the examination of the oral phase of swallowing should use both static digital (still-frame) and dynamic (real-time) assessment methods, as well as expert reviewers. PMID- 11857514 TI - Small focal areas of acoustic shadowing in the breast. PMID- 11857515 TI - Intermittent drop in blood flow velocity in the internal carotid artery and its branches. AB - Extracranial Doppler sonography and transcranial Doppler sonography (TCD) allow the assessment and monitoring of hemodynamic and embolic events in cerebrovascular diseases. We describe an unusual hemodynamic phenomenon in a patient with intracranial carotid siphon stenosis and no clinical symptoms of stenosis. TCD examination suggested and angiography confirmed stenosis of the left internal carotid artery siphon. TCD examination revealed a sudden, intermittent drop in blood flow velocity in both the prestenotic and poststenotic segments of the internal carotid artery, whereas cardiac hemodynamic parameters were unaffected. Embolic signals were detected in the poststenotic vessels only. We speculate that such sonographic findings may be caused by intermittent vessel occlusions due to the reversible displacement of an intraluminal thrombus in relation to the cardiac cycle. PMID- 11857517 TI - Sonographic features of polyorchidism. AB - We describe a case of polyorchidism diagnosed sonographically. Palpation revealed a nontender oval mass, about 1 cm large, inferior to the left testis. Sonography identified the mass as an accessory testis that measured 1.0 x 0.8 x 0.6 cm and had its own epididymis. The echotexture in the accessory testis was similar to that in the normal testes, and color Doppler sonography demonstrated that it had normal blood supply. MRI confirmed these findings. High-resolution sonography is an effective, noninvasive means of accurately diagnosing polyorchidism without the need for exploratory surgery. PMID- 11857516 TI - Prenatal sonographic diagnosis of diastrophic dwarfism. AB - A healthy 27-year-old pregnant woman underwent sonographic examination because her uterine size was large for 20 weeks' menstrual age. Sonograms showed short fetal limbs with hitchhiker thumbs and toes, thoracic scoliosis, clubbed feet, and polyhydramnios. The ossification of all bony structures appeared normal, and there was no evidence of fractures. On the basis of these sonographic findings, we diagnosed skeletal dysplasia and short-limbed dwarfism, most likely diastrophic dwarfism. We counseled the parents, and the pregnancy was continued. At 37 weeks menstrual age, the patient vaginally delivered a male infant that weighed 2,560 g. The infant survived with respiratory support during his first few days of life. Postnatal physical and radiologic examinations confirmed the prenatal diagnosis of diastrophic dwarfism. Sonography is the modality of choice for prenatal detection of diastrophic dwarfism. PMID- 11857518 TI - Intrarenal arteriovenous fistula causing a "pseudonutcracker effect". AB - We report a case in which an intrarenal arteriovenous fistula caused the nutcracker effect. Color Doppler sonography of the left renal artery showed a peak systolic velocity of 150 cm/second and renoaortic ratio of 2.1. The left renal vein was significantly dilated, and the adjacent renal artery was kinked. A stenosis in the left renal vein at the aorto-mesenteric bifurcation was demonstrated, with a maximum velocity of 201 cm/second. Renal Doppler evaluation showed an area of high-velocity, low-resistance arterial flow consistent with an arteriovenous fistula in the inferior half of the left kidney. Angiography confirmed the fistula. The fistula was successfully occluded by coil embolization. Follow-up sonography showed almost complete obliteration of the fistula, regression of the renal vein dilatation, and a reduction in renal venous and arterial flow velocities. Given the reversibility of the fistula's effects, we suggest the term "pseudonutcracker effect" to describe this case. PMID- 11857519 TI - Hydatid cyst of the liver presenting as an inferior vena cava obstruction. AB - A 48-year-old woman presented with clinical features suggestive of an obstruction of the inferior vena cava (IVC). Abdominal sonography showed a 15- x 14-cm cystic mass involving the medial segment of the left hepatic lobe and the anterior segment of the right hepatic lobe. We visualized multiple intracystic floating membranes, a detached endocyst, and a daughter cyst within the main cyst. The cyst was compressing and effacing the intrahepatic portion of the IVC and the right atrium. Sonography of the right and left hepatic veins showed blood flow in the normal direction, but spectral analysis showed that the flow was continuous, possibly owing to extrinsic compression of the IVC at the site where the hepatic veins drained into the IVC. The patient responded well to percutaneous treatment of the hydatid cyst. This case represents a rare presentation of a hydatid cyst of the liver. PMID- 11857520 TI - Efficacy of lamivudine in HBeAg-negative chronic hepatitis B. AB - Hepatitis B e antigen (HBeAg)-negative chronic hepatitis B results from infection with hepatitis B virus mutants unable to produce HBeAg. It accounts for 7-30% of patients with chronic hepatitis B virus (HBV) worldwide, with the highest rates reported for Mediterranean Europe and Asia. Interferon (IFN) is currently the only approved therapy for these patients, but it has an unfavorable tolerance profile and limited efficacy. Studies show that responses to IFN are lower in HBeAg-negative than in HBeAg-positive patients; joint HBV DNA loss/ALT normalization is obtained in 38-59% of HBeAg-negative patients treated for 4-24 months with a high rate of virological relapse (54-87%), at 6-24 months posttreatment. Lamivudine is a nucleoside analogue with potent antiviral properties against HBV. Studies show that response rates in HBeAg-negative and HBeAg-positive patients are equivalent. After 12 months of treatment, 65-96% of HBeAg-negative patients have joint HBV DNA loss/ALT normalization, although 48 74% of patients relapse within 1 year posttreatment. 60% of patients have histological improvement after 12 months of treatment. Lamivudine is well tolerated with a safety profile equivalent to that of placebo. The incidence of YMDD variants increases with extended lamivudine treatment, present in up to 57 64% of patients after 2 years. Their clinical impact is unclear; some studies show breakthrough infection associated with their emergence, whereas other studies show maintained response to lamivudine. Lamivudine has benefits over IFN in its safety and efficacy profile in this patient group. Extended lamivudine treatment beyond 2 years is an option, but further investigation is required to define stopping criteria and the impact of YMDD variants. PMID- 11857521 TI - Clinical and immunological efficacy of intradermal vaccine plus lamivudine with or without interleukin-2 in patients with chronic hepatitis B. AB - To evaluate therapeutic immunostimulation nine chronic hepatitis B patients received six monthly intradermal vaccinations with HBsAg in combination with daily lamivudine. Another five patients received six doses of the vaccine and daily lamivudine together with daily Interleukin-2 (IL-2) s.c. within 3 months in an open-labeled trial. Clinical efficacy was assessed by alanine transaminase levels and HBV serology. The induction of specific T and B cell responses was analyzed serially by 3H-thymidine uptake, ELISA and ELISPOT assays. After the therapy was stopped, seven of nine vaccine/lamivudine and two of five vaccine/lamivudine/IL-2 recipients did not have detectable HBV DNA. Four complete responders cleared the virus and had normalized ALT levels, however, one of these patients showed transient disease reactivation followed by spontaneous viral clearance and normal ALT five months later. Low frequencies of anti-HBs producing B cells and HBV specific T helper cells secreting predominantly interferon-gamma were induced by i.d. vaccine therapy. The ELISPOT technique demonstrated transient induction of HBV peptide specific cytotoxic T cells in seven HLA-A2 positive chronic HBV carriers. The preliminary data from this study demonstrate that the HBV surface antigen vaccine in combination with antiviral or immunomodulating drugs induced antiviral immune responses and consequently viral elimination may be achieved in patients with unfavorable prognosis. PMID- 11857522 TI - Phylogenetic analysis indicates transmission of hepatitis C virus from an infected orthopedic surgeon to a patient. AB - During recent years, a controversial discussion has emerged in the medical community on the real number and possible public health implications of hepatitis C virus (HCV) transmissions from infected medical staff to susceptible patients. We report here on molecular virological and epidemiological analyses involving 229 patients who underwent exposure-prone operations by an HCV-infected orthopedic surgeon. Of the 229 individuals affected, 207 could be tested. Three were positive for HCV antibodies. Molecular and epidemiological investigation revealed that two of them were not infected by the surgeon. The third patient, a 50-year-old man, underwent complicated total hip arthroplasty with trochanteric osteotomy. He harbored an HCV 2b isolate that in phylogenetic analysis of the hypervariable region 1 (HVR 1) was closely related to the HCV strain recovered from the infected surgeon, indicating that HCV-provider-to-patient transmission occurred intraoperatively. To our knowledge, this is the first documented case of HCV transmission by an orthopedic surgeon. The recorded transmission rate of 0.48% (95% confidence interval: 0.09-2.68%) was within the same range reported previously for the spread of hepatitis B virus during orthopedic procedures. Since the result of our investigation sustains the notion that patients may contract HCV from infected health-care workers during exposure-prone procedures, a series of further retrospective exercises is needed to assess more precisely the risk of HCV provider-to-patient transmission and to delineate from these studies recommendations for the guidance and management of HCV-infected medical personnel. PMID- 11857523 TI - Lack of susceptibility of Chacma baboons (Papio ursinus orientalis) to hepatitis C virus infection. AB - The main reason to ascertain whether baboons are susceptible to infection with hepatitis C virus (HCV) is the need to replace chimpanzees, which are endangered, as an animal model for undertaking research into the biology and host-virus interactions of HCV, and for developing a vaccine against this virus. A second reason is that baboons are a possible source of xenografts for human liver transplantation. We inoculated serum containing HCV into four Chacma baboons and monitored them for 52 weeks for evidence of infection. Serum was tested for antibody to HCV, HCV RNA, and aminotransferase concentrations at 2-week intervals for 26 weeks and thereafter at 4-week intervals. Liver tissue was examined at 28 and 52 weeks for histopathological changes and viral RNA, and at 52 weeks for viral particles using electron microscopy. Reverse transcription-polymerase chain reaction assay was used to detect HCV RNA, and the results were confirmed by Southern hybridization. Serum aminotransferase concentrations remained within the normal range and liver histology was normal during the follow-up period. Passive transmission of anti-HCV to the baboons was observed during the first 4 weeks. HCV RNA was not detectable in any serum or liver sample and electron microscopy failed to reveal viral particles in liver tissue. In conclusion, we did not find Chacma baboons to be susceptible to infection with HCV, although we cannot deny that in an immunosuppressed liver transplant recipient, infection of a baboon xenograft might occur. Another animal model for HCV infection must be sought. PMID- 11857524 TI - Evolution of viral quasispecies in four dominant HlA-A2 restricted T cell epitopes is not a major reason for viral persistence in interferon-treated patients with chronic hepatitis C. AB - In most patients, chronic hepatitis C virus (HCV) infection persists despite antiviral treatment with interferon-alpha (IFN-alpha) and ribavirin. The aim of the study was to determine whether HCV could evade cellular immune responses through mutations within T cell epitopes. Viral sequences flanking four major CTL epitopes within the HCV core and envelope regions were analyzed by PCR amplification, cloning and sequencing in seven HLA-A2 positive HCV patients before, during and after antiviral therapy. In addition, cytotoxic T lymphocyte precursor (CTLp) frequencies specific to these epitopes were quantitated by ELISPOT. A total of 13 coding mutations were observed among 650 cloned and sequenced PCR products under or post IFN treatment but no clear selection of viral variants. In detail, the diversity of quasispecies in the two core epitopes remained fairly stable over time despite variable CTLp induction in some individuals. The overall mutation rate in the two envelope epitopes was higher but there was no correlation with specific CTLp frequencies. In conclusion, although evolution of the viral quasispecies during and after antiviral therapy was demonstrated, immune evasion by epitope specific mutations seemed to be not common in interferon nonresponders because the viral complexity did not increase. PMID- 11857525 TI - Factors predictive of response to interferon-alpha therapy in hepatitis C virus type 1b infection. AB - Interferon-alpha (IFN-alpha) has been used to treat hepatitis C Virus (HCV) induced infection but has been effective in only about half of all patients. It is suggested that the different responses to IFN-alpha treatment in HCV infection may be influenced by HCV genotypes, HCV RNA titer at the beginning of IFN-alpha therapy, and the sequences of the interferon sensitivity determining region (ISDR). However, there have also been reports showing that these have no relation to an IFN-alpha effect. In a previous study, it was found that the nucleotide sequence variation in the hypervariable region (HVR) 1 of the HCV could predict the effect of IFN-alpha. In the present investigation, an attempt was made to determine the predictive factors of IFN-alpha therapy. Twenty-six patients with HCV infection were treated with IFN-alpha. Among these, 13 patients recovered after 3 to 6 months of IFN-alpha treatment, although the other 13 patients showed no response after 6 months of treatment with IFN-alpha. In order to determine the predictive factors of IFN-alpha therapy, the ALT levels, HCV genotypes, HCV serum titer, and the quasispecies of HVR 1 were compared between responders and non responders. It is suggested that the variation in the HVR 1 and HCV serum titer can be used to predict the effect of IFN-alpha. PMID- 11857526 TI - Spread of hepatitis E virus among different-aged pigs: two-year survey in Taiwan. AB - Swine are reservoirs of hepatitis E virus (HEV). In this study, a 2-year survey of HEV in feces and sera of swine was conducted to determine if: 1) HEV has circulated among pigs for some time in Taiwan; 2) the spread of HEV among different-aged pigs; and 3) there exists HEV strains possibly imported through trading. From 1998-2000, 521 serum samples and 54 fecal specimens from pigs were examined by reverse transcription polymerase chain reaction. None of the 11 pigs in suckling stage (< 2 months) were serum HEV RNA positive. The highest viremia rate (4.5%) was in pigs of 2 months age, followed by 1.2% and 1.8% in pigs of growing (3-4 months) and finishing stages (5-6 months), and none in pigs older than 6 months. Viremia showed little variation in different years and areas. None of the 20 fecal samples from pigs in suckling stage were HEV RNA positive, whereas 9% of the 34 samples from pigs in growing or finishing stages were positive. Most swine HEV isolates in Taiwan clustered within the genotype 4, whereas the three HEV isolates cloned from pigs imported recently from the U.S. belonged to the genotype 3 HEV in the U.S. The results suggest that HEV may infect pigs at an early growing stage and spread unnoticed among pigs and possibly across countries through trading. PMID- 11857527 TI - Case report: human herpesvirus 7 associated fatal encephalitis in a peripheral blood stem cell transplant recipient. AB - Previous studies have suggested a neuroinvasive and neuropersistent potential of human herpesvirus 7 (HHV-7). In this report, a case of fatal encephalitis is described and its association with HHV-7 infection is discussed. An 8-year-old girl received a peripheral blood stem cell transplant for relapsed acute lymphoblastic leukaemia. The post-transplant period was uneventful and a course of intrathecal chemotherapy was given on Day-30. On Day-41, she developed acute encephalopathy with diplopia and nystagmus. She ran a rapid downhill course and succumbed despite antiviral treatment. The only positive pathological finding was the multiple microscopic foci of haemorrhage associated with neuronal degeneration detected in the brain stem. All microbiological investigations were negative, except for the presence of HHV-7 DNA in cerebrospinal fluid and brain stem tissue samples. PMID- 11857528 TI - Latent infection of human herpesvirus 6 in astrocytoma cell line and alteration of cytokine synthesis. AB - In order to study the pathogenesis of HHV-6 infection in central nervous system disorders, U251 cell line was infected with freshly isolated variant B HHV-6. Although IEA/ex 3 antigen (immediate early protein) was detected in infected cell nuclei, neither the presence of OHV-3 antigen (late antigen) nor production of infectious virus was demonstrated. These results indicate that abortive infection was established in the cells. After viral infection, the viral genome copy in the infected cells gradually decreased and became stable after 6 cell passages. At that point, HHV-6 gene expression was restricted to only 2 immediate early genes (U90 and U94). However, 12-O-tetra-decanoyl (TPA) treatment induced transcription of other genes (U31 and U39) by the 10th cell passage, indicating HHV-6 reactivation. Moreover, production of two proinflamatory cytokines (IL-6 and IL 1beta) was up-regulated by the presence of the HHV-6 genome and TPA-induced activation of the viral transcripts. PMID- 11857529 TI - Multiplex detection of herpesviruses in tear fluid using the "stair primers" PCR method: prospective study of 93 patients. AB - Human herpesviruses can infect the eye and be excreted subsequently in tears. The aim of the present study was to use a multiplex PCR to detect herpesviruses (HSV 1, -2, VZV, CMV, EBV, HHV-6) in tears from normal subjects and from patients with pathological conditions (acute herpes, zoster, papillary conjunctivitis, and dry eye). Schirmer test strips were used to collect tear fluid from 93 patients, sampling both eyes. DNA was then extracted from the 186 samples by chromatography, and viral DNA amplified using a commercialised multiplex "stair primer" method. Thirty-four samples (18.3%) contained Taq inhibitors. The multiplex test gave positive results for HSV and VZV in tear fluid from patients with acute dendritic keratitis (3 patients) and acute ocular zoster (4 patients) and was, therefore, considered effective in testing samples from patients with acute lesions. HSV-1 and HSV-2 were found in two samples from patients with metaherpetic corneal scarring. Among 28 cases of dry eye, two were positive for HHV-6, the latter being associated with EBV in one patient. HHV-6 was also found in 4 out of 54 cases of papillary conjunctivitis. This raised occurrence of HHV-6 in dry eye or papillary conjunctivitis, suggests new clinical patterns for HHV-6 latency or reactivation. Detection of EBV in 1 out of 80 healthy eyes confirms previous evidence that lacrimal glands constitute potentially a site for latent phase EBV. PMID- 11857530 TI - Detection and typing of respiratory adenoviruses in a single-tube multiplex polymerase chain reaction. AB - A multiplex polymerase chain reaction (PCR) assay that is capable of detecting and typing six serotypes of respiratory adenovirus (Ad) was developed, using multiple sets of type-specific primers. The detection of each different serotype depended on distinguishing different numbers and sizes of amplification products on agarose gels following PCR. The multiplex PCR was tested with 26 clinical Ad isolates and other respiratory viruses including influenza viruses, parainfluenza viruses, and respiratory syncytial viruses as well as respiratory bacterial pathogens such as Chlamydia pneumoniae, Streptococcus pneumoniae, Haemophilus influenzae, and Mycoplasma pneumoniae. The multiplex PCR for the detection and typing of Ads gave an excellent correlation with the results by conventional typing with type-specific antisera. This assay may serve as a rapid means of confirming Ad with simultaneous serotype identification of the isolates. It will also have relevance as an adjunctive tool to conventional serotyping for diagnostic and epidemiological purposes. PMID- 11857531 TI - Development and evaluation of an internally controlled semiautomated PCR assay for quantification of cell-free cytomegalovirus. AB - Quantification of circulating human cytomegalovirus (HCMV) is useful in clinical contexts such as virological surveillance of bone marrow transplant recipients and monitoring of antiviral therapy. This report describes an internally controlled, quantitative, semiautomated, HCMV genome assay that was developed primarily to measure HCMV DNA in the plasma of severely leucopaenic patients. It exhibits greater sensitivity, wider dynamic range and higher sample throughput than a number of previously described commercial and "in-house" assays. Viral DNA extraction from EDTA plasma samples was automated using a BioRobot 9604 (Qiagen). HCMV strain AD169 was used to prepare a calibration curve and murine cytomegalovirus (MCMV) strain Smith was added as internal control to all calibration standards and test samples. Amplification was performed using a set of primers based on the HCMV UL50 region, capable of amplifying both human and murine CMV. The yield of biotinylated polymerase chain reaction (PCR) products was estimated using HCMV-specific and MCMV-specific enzyme-labelled probes and automated chemiluminescence detection. Log-transformed HCMV-to-MCMV signal ratios were calculated and used for quantification of test samples against simultaneously extracted MCMV-spiked calibration standards. Evaluation of the assay sensitivity by Probit analysis demonstrated a 95% probability of detection at 100 HCMV genomes per ml of plasma; the dynamic range was shown to be > or = 4 log(10). A total of 315 samples from 61 bone marrow transplant patients were analysed by both the quantitative PCR (qPCR) and by a previously validated nested nonquantitative PCR (NQPCR). A high level of concordance (90%) was observed between the two assays, although the qPCR assay exhibited slightly greater sensitivity. PMID- 11857532 TI - Single rapid TaqMan fluorogenic probe based PCR assay that detects all four dengue serotypes. AB - Public health laboratories require rapid diagnosis of dengue outbreaks for application of measures such as vector control. We have developed a rapid single fluorogenic probe-based polymerase chain reaction assay for the detection of all four dengue serotypes (FUDRT-PCR). The method employs primers and probe that are complementary to the evolutionarily conserved 3' untranslated region of the dengue genome. The assay detected viral RNA of strains of all four dengue serotypes but not of the flaviviruses Japanese encephalitis virus, Murray Valley encephalitis virus, Kunjin, Stratford, West Nile, Alfuy or Yellow fever. When compared to an existing nested-PCR assay for the detection of dengue on clinical samples, FUDRT-PCR detected dengue 1 (100%, n=14), dengue 2 (85%, n=13), dengue 3 (64%, n=14) and dengue 4 (100%, n=3) with the indicated sensitivities. FUDRT-PCR enables diagnosis of acute dengue infection in four hours from sample receipt. In addition, a single-test procedure should result in a reduction in the number of tests performed with considerable cost savings for diagnostic laboratories. PMID- 11857533 TI - Detection and identification of dengue-1 virus in clinical samples by a nested PCR followed by restriction enzyme digestion of amplicons. AB - Dengue viruses cause a disease with clinical findings ranging from asymptomatic infections to severe manifestations, characterised by haemorrhage and shock and known as dengue haemorrhagic fever/dengue shock syndrome. Since this fever and syndrome usually results from sequential infections by distinct dengue serotypes, rapid detection and identification of dengue viruses circulating in endemic areas are important to implement control measures, and ultimately to avoid secondary infections that could result in dengue haemorrhagic fever/dengue shock syndrome. A nested-PCR was developed followed by restriction enzyme (Kpn I) digestion of the amplicons to differentiate dengue-1 from dengue-2. Seventy-five IgM containing samples collected from 2 to 17 days after the beginning of the symptoms were examined. These samples were submitted to nested-PCR amplification, the amplicons were digested with Kpn I, and the results compared to virus isolation in C6/36 cells and to results obtained by the standard PCR. Out of 75 tested samples, virus was isolated from 2 (2.6%), 17 (22.7%) were positive by the regular PCR protocol, and 58 (77.3%) were positive by nested-PCR. All of the amplicons digested by Kpn I identified dengue-1 virus as the infecting strain. These results indicate that the nested-PCR provided a high yield of dengue genome amplification even in the presence of IgM antibodies, and restriction enzyme digestion defined rapidly the circulating serotype. Therefore, the combination of these techniques may be useful to rapidly identify dengue viruses in countries where dengue-1 and dengue-2 circulates, and this approach can also be applied to the other two serotypes. PMID- 11857534 TI - Population-based type-specific prevalence of high-risk human papillomavirus infection in middle-aged Swedish women. AB - Human papillomavirus (HPV) DNA testing can be used to identify women at risk of the development of cervical cancer. The cost-effectiveness of HPV screening is dependent on the type-specific HPV prevalence in the general population. The present study describes the prevalence and spectrum of high-risk HPV types found in a large real-life population-based HPV screening trial undertaken entirely within the cervical screening program offered to middle-aged Swedish women. Cervical brush samples from 6,123 women aged 32-38 years were analyzed using a general HPV primer (GP5+/6+) polymerase chain reaction-enzyme immunoassay (PCR EIA) combined with reverse dot-blot hybridization for confirmation and HPV typing by a single assay. In this study, 6.8% (95% CI 6.2-7.5) (417/6,123) were confirmed as high-risk HPV positive. Infections with 13 different high-risk HPV types were detected, of which HPV 16 was the most prevalent type (2.1%; 128/6,123), followed by HPV 31 (1.1%; 67/6,123). Any one of the HPV types 18, 33, 35, 39, 45, 51, 52, 56, 58, 59, or 66 was detected in 3.6% (223/6,123) of the women. Infection with two, three, and five types simultaneously was identified in 32, 5, and 1 women, respectively. The combination of PCR-EIA as a screening test and reverse dot-blot hybridization as a confirmatory test, was found to be readily applicable to a real-life population-based cervical screening. The type specific HPV prevalence found support in previous modeling studies suggesting that HPV screening may be a favorable cervical screening strategy. PMID- 11857535 TI - Characteristics of Puumala and Dobrava infections in Croatia. AB - In this study, two different hantaviruses, Puumala virus (PUUV) and Dobrava virus (DOBV), were demonstrated for the first time to coexist and cause hemorrhagic fever with renal syndrome (HFRS) in Croatia. Phylogenetic analysis showed some differences among the nucleotide sequences of PUUV originating from Dinara mountain, which was more closely related to Austrian PUUV than other Croatian PUUV from Mala Kapela mountain. More consistency was found among the Croatian DOBV. HFRS was verified in 85 of 201 suspected cases recorded in 1995 during the largest HFRS outbreak in Croatia. Most of these cases were soldiers. With the exception of the coastal region and islands, all of Croatia was found to be an area endemic for HFRS. A statistically significantly higher proportion of DOBV infected patients had acute renal failure, visual disturbance, severe thrombocytopenia, and elevated levels of nonsegmented leukocytes, creatine, and total bilirubin. The prevalence of gastrointestinal and electrocardiography disorders also was greater in DOBV-infected patients. Interestingly, significantly more PUUV-infected patients had elevated systolic blood pressure on admission to the hospital. Further prospective studies are necessary to shed more light on differences in HFRS severity associated with PUU and DOB viruses. PMID- 11857536 TI - Effect of lack of Interleukin-4, Interleukin-12, Interleukin-18, or the Interferon-gamma receptor on virus replication, cytokine response, and lung pathology during respiratory syncytial virus infection in mice. AB - RSV is an important cause of bronchiolitis in infants. Immunopathology may play a role in RSV-induced bronchiolitis and severe RSV-induced disease has been associated with a Th2 type immune response. The aim of the study was to identify cytokine pathways that are crucial in influencing RSV-induced disease. For that purpose we inoculated IFNgammaR-/-, IL-12-/-, IL-18-/-, or IL-4-/- mice with RSV. We observed that an RSV infection resulted in a predominant Th1 cytokine response associated with slight bronchiolitis and alveolitis. Pulmonary histopathology was only aggravated in IFN R-/- mice, characterised by eosinophilic influx around the bronchioles. Despite subtle changes in cytokine expression, no differences in histopathology were observed in IL-12-/- and IL-18-/- mice. Deficiency of IL-4 has no effect on RSV-induced Th1 cytokines and pulmonary histopathology. IFNgamma receptor deficiency during primary RSV infection resulted in a disturbed Th1 response based on increased IL-4, IL-5, and IL-13 expression and the presence of eosinophils in the lungs. It is concluded that IFNgamma signalling is required for a pronounced Th1 response to RSV while IL-12 and IL-18 are not. A shift in the balance between Th1 and Th2 towards a Th2 response induced by missing IFNgamma signalling leads to aggravated pulmonary pathology. This is not caused by enhanced viral load. PMID- 11857537 TI - TTV infection and its relation to serum transaminases in apparently healthy blood donors and in patients with clotting disorders who have been investigated previously for hepatitis C virus and GBV-C/HGV infection in Belgium. AB - A novel DNA virus, TT virus (TTV), has been proposed as a possible etiologic agent for non A-E hepatitis. The aim of the present study was to determine the prevalence of TTV infection using PCR in healthy blood donors and in patients with clotting disorders who have been investigated previously for GBV-C/HGV and HCV infection in Belgium. In this study, PCR using primers proposed by Takahashi et al. [(1998) Hepatology Research 12:233-239] proved far more sensitive than those used by Okamoto et al. [(1998) Journal of Medical Virology 56:128-132]. The sequence of the PCR products showed 87% identity to the published sequence. TTV was present in 29.7% of healthy blood donors, a figure intermediate between the low rate of infection observed in Scotland and the high rates in the Far East. TTV was detected in 46.5% of 127 patients studied with clotting disorders as compared to 79.5% for HCV and 11.8% for GBV-C/HGV infection. However, there was no impact on the level of serum transaminases. Treatment with interferon for HCV infection co-infected with TTV suppressed temporarily serum TTV DNA. Therefore, it was concluded that TTV DNA is detected frequently in serum of healthy blood donors in Belgium and more often in patients with clotting disorders. TTV does not cause liver disease or contribute to the severity of liver disease. PMID- 11857538 TI - Detection of varicella-zoster virus DNA in throat swabs of patients with herpes zoster and on air purifier filters. AB - Zoster patients are considered to be less contagious than those with varicella because their infectious lesions are localized. However, it is not known when the spread of varicella-zoster virus (VZV) from zoster patients begins, how long it continues, and how far the virus spreads from the zoster patients. Twelve cases of hospitalized zoster patients were studied. The polymerase chain reaction (PCR) was used to detect VZV DNA in samples taken from the surface of their eruptions, throats, and the air purifier filters in their rooms. In all patients, VZV DNA was detected in the samples from eruptions. VZV DNA was detected in 8 of 12 patients from the throats. VZV DNA was detected for 9 of 12 patients from the filter samples. This study shows the possibility of a wide distribution of VZV DNA to the environment from infected patients. VZV may be excreted from cutaneous eruptions or from the throats of patients. PMID- 11857539 TI - Screening of blood from potential organ and cornea donors for viruses. AB - Prospective nucleic acid tests were carried out for human immunodeficiency virus (HIV) and hepatitis C virus (HCV) using the COBAS Amplicor HIV-1 and HCV tests (Roche Diagnostics, Meylan, France) on potential organ (n=113) and cornea (n=368) donors in France to evaluate their performance and suitability for use as a complement to routine serological tests. Blood samples were collected from organ donors with preserved cardiac function after verification of cerebral death. Blood samples were collected from cornea donors post-mortem within 48 hr after death. An internal control was added to the samples before extraction to monitor each individual polymerase chain reaction (PCR). The nucleic acid tests were always interpretable in organ donors and negative in all except in 2 anti-HCV positive patients. One had an indeterminate HIV p24 antigen but was negative for HIV RNA. HIV and HCV RNA were not found in cornea donors with a negative serology but indeterminate molecular results were frequent in this group (17.6%). Cornea donors also gave significantly more (14.4%) indeterminate serological results than organ donors (1.8%) (P<0.001). This was due to the poor quality of the blood samples collected post-mortem. However, there was no correlation between indeterminate results of serological and molecular tests. There were 16/19 (84%) indeterminate serological results for HIV and 4/4 (100%) for HCV that were negative by PCR. Thus, nucleic acid tests could be useful for qualifying a donor whose serological results are indeterminate. The extraction procedures on post mortem specimens and/or blood collection must be changed to improve the performance of nucleic acid tests. PMID- 11857540 TI - Inhalation anesthetic-induced neuroinvasion by an attenuated strain of West Nile virus in mice. AB - There are contradictory reports regarding the effects of inhalation anesthetics on the immune system. Measurable immune responses have been studied in vitro, but little is known about the in vivo effects in the intact organism. We used an attenuated, non-neuroinvasive, nonlethal strain of the encephalitic West Nile virus, termed WN-25, which can become lethal in combination with environmental stressors, to study possible modulatory immune effects of inhalation anesthetics in mice. Both single short-term exposure and repeated exposure to halothane and nitrous oxide were studied. Exposure to 30% CO2 served as a positive control. Mortality, brain invasion, spleen weight, and antiviral antibodies served as the experimental endpoints. Halothane and nitrous oxide led to viral brain invasion, increased mortality, and suppressed immune response in a concentration- and time dependent manner. Repeated exposures had a cumulative effect. Assessment of the stability of the viral attenuation did not demonstrate any alteration in the character of the virus, suggesting an increased access to the brain by inhalation anesthetics that led to the fatal encephalitis. These findings may be of special concern to populations at risk, such as operating room staff and patients undergoing general anesthesia in endemic areas of encephalitic virus species, in which subclinical infection may develop into an overt disease. PMID- 11857541 TI - Maternal periconceptional vitamin use, genetic variation of infant reduced folate carrier (A80G), and risk of spina bifida. AB - Women who consume folic acid in early pregnancy reduced their risks for delivering offspring with neural tube defects (NTDs). The underlying process by which folic acid facilitated this risk reduction is unknown. Investigating genetic variation that influences cellular absorption, transport, and metabolism of folate will help fill this data gap. We focused our studies on a candidate gene that is involved in folate transport, the reduced folate carrier 1 (RFC1). Using data from a California population-based case control interview study (1989 1991 birth cohorts), we investigated whether spina bifida risk was influenced by an interaction between a polymorphism of infant RFC1 at nucleotide 80 (A80G) and maternal periconceptional use of vitamins containing folic acid. Allelic variants of RFC1 were determined by genotyping 133 live-born spina bifida case infants and 188 control infants. The percentages of case infants with the A80/A80, G80/G80, and G80/A80 genotypes were 27.2%, 28.0%, and 44.7%, respectively. The percentages of control infants were similar: 26.1%, 29.3%, and 44.7%. Odds ratios of 1.0 (95% confidence interval 0.5-2.0) for the G80/G80 genotype and 1.1 (0.6-2.0) for the G80/A80 genotype were observed relative to the A80/A80 genotype. Among mothers who did not use vitamins, spina bifida risk was 2.4 (0.8-6.9) for infants with genotype G80/G80 compared to those with A80/A80 genotype. Among mothers who did use vitamins, the risk was 0.5 (0.1-3.1) for infants with the G80/G80 genotype. Although this study did not find an increased spina bifida risk for infants who were heterozygous or homozygous for RFC1 A80G, it did reveal modest evidence for a gene-nutrient interaction between infant homozygosity for the RFC1 G80/G80 genotype and maternal periconceptional intake of vitamins containing folic acid on the risk of spina bifida. PMID- 11857542 TI - Spectrum of the acrocallosal syndrome. AB - Acrocallosal syndrome (ACS) is an autosomal recessive condition, characterized by agenesis of the corpus callosum, pre- and postaxial polydactyly, minor craniofacial anomalies, and, in most patients, severe psychomotor retardation. We here report on three patients with ACS demonstrating a spectrum from mild to severe involvement. Two patients had only mild to moderate mental retardation at the age of 2(1/2) and 4 years, respectively, with surprisingly good speech development. The third patient was severely affected and died at age 7 days because of persistent apnea. All three patients had agenesis of the corpus callosum, and large intracranial cysts, which in the third case was confirmed as a large arachnoid cyst at autopsy. Cranial cysts were also seen in 10/34 published cases of ACS. Thus, intracerebral cysts are a common finding in ACS and may serve in differentiating ACS from Greig cephalopolysyndactyly syndrome. PMID- 11857543 TI - Mutational analysis of the Sonic Hedgehog gene in 220 newborns with oral clefts in a South American (ECLAMC) population. AB - Oral clefts generally have a multifactorial etiology. A number of genes contribute to the formation of the face and palate. Cleft lip and/or palate can occur in pedigrees with autosomal dominant holoprosencephaly due to mutations in Sonic Hedgehog (SHH). In addition, animal models have shown that SHH is involved in face development. We thus examined the human SHH gene in 220 newborn infants with nonsyndromic oral clefts registered by the Estudio Colaborativo Latinoamericano de Malformaciones Congenitas: ECLAMC (Latin American Collaborative Study of Congenital Malformations). We found 15 variant bands in 13 patients with oral clefts, representing five different base changes, all of which were found by sequencing to represent silent polymorphisms. Four occurred in introns. The alteration occurring in an exon, Ser190Ser, may create a consensus sequence for the 3'splice site 6 bp downstream of the original consensus sequence. Thus, we did not identify any clearly disease-causing mutation in SHH in these patients, and conclude that SHH mutations are not a frequent cause of isolated oral clefts in humans. PMID- 11857544 TI - Apparent genotype-phenotype correlation in childhood, adolescent, and adult Chediak-Higashi syndrome. AB - Chediak-Higashi syndrome (CHS) is a rare autosomal recessive disorder characterized by severe immunologic defects, reduced pigmentation, bleeding tendency, and progressive neurological dysfunction. Most patients present in early childhood and die unless treated by bone marrow transplantation. About 10 15% of patients exhibit a much milder clinical phenotype and survive to adulthood, but develop progressive and often fatal neurological dysfunction. Very rare patients exhibit an intermediate adolescent CHS phenotype, presenting with severe infections in early childhood, but a milder course by adolescence, with no accelerated phase. Here, we describe the organization and genomic DNA sequence of the CHS1 gene and mutation analysis of 21 unrelated patients with the childhood, adolescent, and adult forms of CHS. In patients with severe childhood CHS, we found only functionally null mutant CHS1 alleles, whereas in patients with the adolescent and adult forms of CHS we also found missense mutant alleles that likely encode CHS1 polypeptides with partial function. Together, these results suggest an allelic genotype-phenotype relationship among the various clinical forms of CHS. PMID- 11857546 TI - Serious genetic disorders: can or should they be defined? AB - The word "serious" appears in laws and policies regarding genetics services but is not defined. Genetics professionals would most likely be consulted if definitions are made. We surveyed all U.S. board-certified genetics services providers and all members of the European Society of Human Genetics (ESHG), Canadian College of Medical Geneticists (CCMG), and Ibero-American Society of Human Genetics (IASHG), using anonymous, mailed questionnaires. Respondents were asked to list three conditions they considered lethal, three that were serious but not lethal, and three that were not serious. Of 3,317 asked to respond, 1,481 (45%) returned questionnaires. Analysis was limited to responses of the 1,264 (85%) who saw patients. Respondents listed 537 conditions, with extensive overlap between categories; 46% of conditions listed as serious were also listed as not serious and 41% were listed as lethal. Respondents did not want professional societies, laws, or national ethics committees to define serious. They favored individual patients as decision makers, with the help of individual doctors. Their reasons were that genetic disorders vary in expression, individuals perceive disorders differently, professional or legal definitions could be unfair to minority groups, definitions tend to be inflexible when new treatments appear. In the United States, most would not use a definition of serious to limit reproductive services, carrier screening, or neonatal intensive care; outside the United States, more supported limitations. There is not sufficient consensus among experienced genetics professionals to define serious genetic conditions for purposes of law or policy. Responses point to social and economic influences on perceptions of what constitutes a serious genetic disorder. PMID- 11857545 TI - Common mutations of ATP7B in Wilson disease patients from Hungary. AB - Wilson disease (WD) is an autosomal recessive disorder of copper metabolism. The H1069Q mutation in exon 14 of ATP7B is far the most frequent in Wilson patients of European origin. Mutations in exon 8 and 15 are also common among the over 150 described mutations in the WD gene. The aim was to investigate the frequency of these common WD gene mutations in Hungarian patients. A total of 42 patients with WD from 39 Hungarian families were examined. The H1069Q mutation was assessed by a seminested polymerase chain reaction (PCR)-based restriction fragment length polymorphism (RFLP) assay, while mutations in exons 8, 13, 15, and 18 of WD gene were identified by sequencing. In addition, haplotype analysis was performed using three common microsatellite markers (D13S314, D13S301, D13S316). The H1069Q mutation was found in 27 patients (64.3%). Nine patients were H1069Q homozygous. Eighteen patients were H1069Q compound heterozygous, two of them had H1069Q/P969Q and one patient H1069Q/3400delC genotype. In two of the 15 H1069Q-negative patients a novel mutation in exon 13 (T977M) was detected. One H1069Q-negative patient had a mutation in exon 8 (G710S). None of the studied mutations was detected in 12 WD patients. H1069Q-positive patients from various European countries had the same haplotype pattern. The H1069Q point mutation is frequent in Hungarian patients with WD and appears to have originated from a single founder in Eastern Europe. In contrast, mutations in exons 8, 13, 15, and 18 are uncommon in Hungarian WD patients. PMID- 11857547 TI - Pure 6p22-pter trisomic patient: refined FISH characterization and genotype phenotype correlation. AB - First described in 1971, partial trisomy 6p is uncommon and generally secondary to a familial reciprocal translocation. The proximal breakpoint of the reported cases varies from p11 to p25. We here report on a patient with moderate mental retardation, craniofacial and pigmentary anomalies, proteinuria, and hyperglycemia who was found to have a mosaic karyotype 46,X,add(Y)(q12)/45,X. Fluorescence in situ hybridization (FISH) enabled us to identify that the additional material on Yqh derived from 6p and to define the rearrangement as der(Y)t(Y;6)(q12;p22). To the best of our knowledge, this is the first case of trisomy 6p22-pter without an associated deleted segment; the second breakpoint of the rearrangement is in Yqh. Precise mapping of the centromeric breakpoint of the trisomic 6p segment allowed a more convincing correlation between partial 6p trisomy and clinical phenotype to be addressed. In particular, the proteinuria often observed in 6p trisomic patients could be assigned to the 6p22-6pter region. PMID- 11857548 TI - Two siblings with early onset fetal akinesia deformation sequence and hydranencephaly: further evidence for autosomal recessive inheritance of hydranencephaly, fowler type. AB - We report a 13-week-old female fetus with early onset fetal akinesia deformation sequence (FADS) and hydranencephaly. In a previous pregnancy, the same ultrasonographic findings were noted at 13 weeks. Fetopathological examination of both female fetuses confirmed FADS with severe arthogryposis, multiple pterygia, and muscular hypoplasia. Neuropathological examination showed massive cystic dilatation of the cerebral ventricles (hydranencephaly) with calcification of the basal ganglion and brain stem and a proliferative vasculopathy throughout the central nervous system. The findings in the two female siblings document the earliest echographic diagnosis of hydranencephaly, Fowler type, and this observation further supports autosomal recessive inheritance of this distinct type of hydranencephaly. PMID- 11857549 TI - Combination of diaphragmatic eventration and microphthalmia/anophthalmia is probably nonrandom. AB - Two sporadic cases of eventration of the diaphragm are reported; one had bilateral colobomatous microphthalmia and the other had anophthalmia. Absence of polydactyly and presence of eventration rather than diaphragmatic hernia helped to exclude Fryns syndrome. These cases together with published cases with overlapping features support the thesis that this combination of defects is nonrandom and of heterogeneous cause. Some cases are due to a pleiotropic gene defect. In other cases, a polytypic developmental field involving an unknown developmental cascade common to the eye and diaphragm may provide a basis for the combination. PMID- 11857550 TI - Hydrocephalus and intestinal aganglionosis: is L1CAM a modifier gene in Hirschsprung disease? AB - Congenital hydrocephalus associated with aqueductal stenosis and/or agenesis of the corpus callosum has been described in newborn males with mutations in L1CAM, a gene that encodes a neural cell adhesion molecule. These males usually have severe mental retardation and may have spastic paraplegia and adducted thumbs. In contrast, Hirschsprung disease, or absence of ganglion cells in the distal gut, has rarely been described in such individuals. We report a male infant who had severe hydrocephalus identified in the prenatal period with evidence of aqueductal stenosis and adducted thumbs at birth. He developed chronic constipation, and rectal biopsy confirmed the diagnosis of Hirschsprung disease. Molecular testing of the L1CAM gene revealed a G2254A mutation, resulting in a V752M amino acid substitution. A common polymorphism in RET, but no mutation, was identified. Our patient represents the third example of coincident hydrocephalus and Hirschsprung disease in an individual with an identified L1CAM mutation. We hypothesize that L1CAM-mediated cell adhesion may be important for the ability of ganglion cell precursors to populate the gut, and that L1CAM may modify the effects of a Hirschsprung disease-associated gene to cause intestinal aganglionosis. PMID- 11857551 TI - Progressive cerebral edema associated with high methionine levels and betaine therapy in a patient with cystathionine beta-synthase (CBS) deficiency. AB - Cystathionine beta-synthase (CBS) deficiency, the most common form of homocystinuria, is an autosomal recessive inborn error of homocysteine metabolism. Treatment of B6-nonresponsive patients centers on lowering homocysteine and its disulfide derivatives (tHcy) by adherence to a methionine restricted diet. However, lifelong dietary control is difficult. Betaine supplementation is used extensively in CBS-deficient patients to lower plasma tHcy. With betaine therapy, methionine levels increase over baseline, but usually remain below 1,500 micromol/L, and these levels have not been associated with adverse affects. We report a child with B6-nonresponsive CBS deficiency and dietary noncompliance whose methionine levels reached 3,000 micromol/L on betaine, and who subsequently developed massive cerebral edema without evidence of thrombosis. We investigated the etiology by determining methionine and betaine metabolites in our patient, and several possible mechanisms for her unusual response to betaine are discussed. We conclude that the cerebral edema was most likely precipitated by the betaine therapy, although the exact mechanism is uncertain. This case cautions physicians to monitor methionine levels in CBS deficient patients on betaine and to consider betaine as an adjunct, not an alternative, to dietary control. PMID- 11857552 TI - Smith-Lemli-Opitz syndrome: new mutation with a mild phenotype. AB - Smith-Lemli-Opitz syndrome (SLOS) (Online Mendelian Inheritance in Man, OMIM, 2001, http://www.ncbi.nlm.nih.gov/omim/ for SLOS, MIM 270400) is an autosomal recessive disorder of cholesterol biosynthesis caused by mutations of the 3beta hydroxysterol Delta(7)-reductase gene, DHCR7. We report on a female infant with an exceptionally mild phenotype of SLOS, in whom molecular studies identified a new mutation in DHCR7. The proposita initially presented with feeding difficulties, failure to thrive, hypotonia, mild developmental delay, and oral tactile aversion. She had minor facial anomalies and 2-3 syndactyly of her toes in both feet. The plasma cholesterol was borderline low at 2.88 mmol/L (normal 2.97-4.40 mmol/L). Elevated plasma 7-dehydrocholesterol level of 200.0 micromol/L confirmed the clinical diagnosis of SLOS. Molecular analysis demonstrated compound heterozygosity for IVS8-1G -->C and Y280C, a new missense mutation in DHCR7. Since the other mutation in this patient is a known null mutation, this newly discovered mutation is presumably associated with significant residual enzyme activity and milder expression of clinical phenotype. PMID- 11857553 TI - Craniofacial anomalies, deafness, brachydactyly, short stature, and moderate mental retardation due to a cryptic 6p;11q translocation. AB - Monozygotic twin brothers are described who share clinical features which include: moderate mental retardation, short stature, macrocephaly, frontal bossing, ptosis, low-set ears, brachydactyly, 5th fingers clinodactyly, single palmar creases, cryptorchidism, and prelingual sensorineural deafness. One of the twins presented with mild cardiac dilatation and died at age 3(1/2) from cardiac arrest during an episode of acute respiratory infection. While chromosome analyses performed for both twins on peripheral blood showed apparently normal karyotypes, screening for all telomeric regions on the surviving propositus revealed a combination of partial 6p trisomy and partial 11q monosomy. A balanced reciprocal translocation was found in the father. The phenotype of the twins is most likely related to this cryptic chromosomal rearrangement. The fact that the phenotype in this family partially overlaps with some previously reported phenotypes is discussed. PMID- 11857554 TI - Apolipoprotein B and apolipoprotein E genotypes and sporadic holoprosencephaly. PMID- 11857555 TI - Evidence for genetic heterogeneity in familial isolated patella aplasia hypoplasia. PMID- 11857556 TI - Five additional Costello syndrome patients with rhabdomyosarcoma: proposal for a tumor screening protocol. AB - We report five new cases of rhabdomyosarcoma (RMS) in Costello syndrome. These cases, combined with those previously reported, increase the number of solid tumors to 17 (10 RMSs, 3 neuroblastomas, 2 bladder carcinomas, 1 vestibular schwannoma, 1 epithelioma), in at least 100 known Costello syndrome patients. Despite possible ascertainment bias, and the incomplete identification of all Costello syndrome patients, the tumor frequency could be as high as 17%. This is comparable to the 7-21% frequency of solid tumors in Beckwith-Wiedemann syndrome (BWS), and may justify tumor screening. Based on the recommendations for screening BWS patients, we propose a screening protocol consisting of ultrasound examination of the abdomen and pelvis every 3-6 months until age 8-10 years for RMS and abdominal neuroblastoma; urine catecholamine metabolite analysis every 6 12 months until age 5 years for neuroblastoma; and urinalysis for hematuria annually for bladder carcinoma after age 10 years. These recommendations may need to be modified, as new information becomes available. Potential criticism of the tumor screening protocol concerns the lack of evidence for improved outcome, and possible overestimation of the tumor risk. The ability of RMSs to occur at various sites complicates tumor screening, but 8 of the 10 RMSs in Costello syndrome patients originated from the abdomen, pelvis and urogenital area. Prior diagnosis of Costello syndrome is a prerequisite for the implementation of any screening protocol. The diagnosis of Costello syndrome should also be considered in individuals with RMS and physical findings suggestive of Costello syndrome. PMID- 11857557 TI - Screening for cancer in children with Costello syndrome. PMID- 11857558 TI - Ring chromosome 6 in three fetuses: case reports, literature review, and implications for prenatal diagnosis. AB - Prenatal and postnatal findings in three fetuses with a ring chromosome 6 are presented, and the literature of this rare cytogenetic disorder is reviewed. The described fetuses illustrate the broad spectrum of the clinical manifestation of ring chromosome 6. In one fetus, the disorder was diagnosed incidentally by a routine amniocentesis due to advanced maternal age. The other two fetuses were hydrocephalic and had other congenital anomalies. Remarkably, the ring chromosome 6 tends to disappear in cultured amniotic fluid cells; karyotyping revealed complete or nearly complete monosomy 6. In contrast, the ring was preserved in high proportions of fetal leukocytes. Postnatal growth retardation is the only consistent finding of this chromosomal disorder. Maternal age is not significantly above average. An additional review of 20 literature cases revealed a striking tendency to hydrocephalus, either due to deficient brain growth or secondary to an aqueductal stenosis. Children with hydrocephalus and ring chromosme 6 tend to display facial dysmorphism and may have additional malformations, growth failure, eye anomalies, and seizures. In contrast, there are two reports on children with a ring chromosome 6 who had short stature, normal appearance, and a normal or almost-normal psychomotor development. In such patients at the mild end of the clinical spectrum, the phenotype is basically restricted to what Kosztolanyi. [1987: Hum Genet 75:174-179] delineated as "ring syndrome," comprising "severe growth failure without major malformations, without a specific deletion syndrome, with only a few or no minor anomalies, and mild to moderate mental retardation." This "ring syndrome" is considered to occur independently of the autosome involved in the ring formation. The overall impression from our cases and from the literature review of cases with ring chromosome 6 is that the karyotype-genotype correlation is poor. This makes prognostic counseling of parents difficult and unsatisfactory. Serial targeted ultrasound examinations, especially of the brain, are decisive factors in elucidating the prognosis. PMID- 11857559 TI - Problem behavior in boys with fragile X syndrome. AB - This study examines problem behavior over time in 59 boys with fragile X syndrome (FXS), aged 4-12 years, using the Child Behavior Checklist (CBCL). Approximately 49% of the boys scored within the borderline or clinical range on total problem behavior, while 56-57% scored in the borderline or clinical range on the attention and thought problems subscales, and 26% scored in this range on the social problems subscale. With a mean of 2.5 assessments per child, behavior problems were stable during the 3-year period of study. Total problem behavior was higher for children who displayed autistic behavior, were rated as low in adaptability, had mothers with higher maternal education levels, and were on medication. Mothers with more education also rated their children as having more attention, thought, and total problems. Children taking medication differed from boys who were not taking medication on social problems, but not on attention and thought problems. Low adaptability and more autistic characteristics predicted thought problems. PMID- 11857560 TI - Frequencies of UDP-glucuronosyltransferase 1 (UGT1A1) gene promoter polymorphisms among distinct ethnic groups from Brazil. AB - A polymorphism in the promoter region of the UDP-glucuronosyltransferase 1 (UGT1A) gene is associated with Gilbert syndrome (GS), a benign form of mild unconjugated hyperbilirubinemia. We genotyped 157 individuals from Brazil, comprising 71 Caucasians, 54 African-derived subjects, and 32 Parakana Indians. Frequencies of the alelle (TA)(7) associated with GS found in this study were 0.324, 0.407, and 0.328, respectively. The genotype frequencies differed significantly between Caucasians and African-derived individuals. The high frequencies of (TA)(7) polymorphism among the three groups confirm previous data that this polymorphism is very ancient and appears to be distributed throughout the world. PMID- 11857561 TI - Phenotypic variation in Melnick-Needles syndrome is not reflected in X inactivation patterns from blood or buccal smear. AB - Melnick-Needles syndrome is a rare putative X-linked dominant bone dysplasia. The patients have short stature, characteristic facial features, and a normal intelligence. The skeletal dysplasia includes S-shaped curvature of tubular bones and sclerosis of the base of the skull. The phenotype of affected individuals varies, even within families. This could be related to X chromosome inactivation. We report here on a very mildly affected mother and her two severely affected daughters with characteristic features of Melnick-Needles syndrome. In addition, the two daughters had very similar pigmented nevi on their back. X chromosome inactivation analysis of blood DNA revealed a skewed X inactivation pattern in all three affected females, with the normal X chromosome as the predominating active X chromosome. The X inactivation pattern was similar in buccal smear and blood DNA in the mother and one of the daughters, whereas the other daughter had a skewed pattern in blood only. X chromosome inactivation in blood and buccal smear DNA therefore does not explain the phenotypic variation in this family. The skewed X chromosome inactivation is in agreement with X-linked inheritance of Melnick-Needles syndrome and suggests a critical role of the Melnick-Needles gene in hematopoietic cell proliferation. Clinical evidence indicates that Melnick Needles syndrome is allelic to the otopalatodigital syndromes, which have been assigned to Xq26-28. Haplotype analysis of the X chromosomes in this family was in agreement with the localization of the gene for Melnick-Needles syndrome to Xq25-qtel. PMID- 11857562 TI - Possible association of NTDs with a polyhistidine tract polymorphism in the ZIC2 gene. AB - Neural tube defects (NTDs) and brain malformations represent a common finding in chromosome 13q deletion patients. Hemizygosity for ZIC2, which is located in the 13q32 critical deletion region, results in holoprosencephaly (HPE) in humans, and diminished expression of ZIC2 results in HPE as well as lumbosacral NTDs in mice. Taken together, these observations led us to hypothesize that ZIC2 mutations may be a cause of isolated NTD. To test this, we screened 192 NTD patients for mutations in ZIC2. While we did not find ZIC2 mutations in these patients, we did find some evidence of a possible association between a histidine tract polymorphism in ZIC2 and NTDs. Our sample was too small to reach definitive conclusions, but the evidence is sufficiently intriguing to encourage further research. If this association is confirmed, subtle alterations in ZIC2 activity may confer a risk of NTD. PMID- 11857563 TI - Familial paraganglioma and gastric stromal sarcoma: a new syndrome distinct from the Carney triad. AB - Paragangliomas may be inherited in an autosomal dominant manner either alone (as in PGL1, PGL2, and PGL3 syndromes) or as a component of a multiple tumor syndrome (as in von Hippel-Lindau disease and neurofibromatosis type 1). In this article, we describe 12 patients (7 male and 5 female) with an average age of 23 years from five unrelated families that manifested paraganglioma and gastric stromal sarcoma; the tumors were inherited in an apparent autosomal dominant manner, with incomplete penetrance. Seven patients had paraganglioma, four had paraganglioma and gastric stromal sarcoma, and one had gastric stromal sarcoma. The paraganglioma was multicentric and the gastric stromal sarcoma multifocal. Because of the rarity of gastric stromal sarcoma and its multifocality, the young age of the patients, and the unlikelihood of coincidental co-occurrence of paragangliomas and gastric stromal sarcomas, we suggest that a new syndrome exists with these two main components, a condition that is familial and distinct from the Carney triad. PMID- 11857565 TI - Mutational analysis of the luteinizing hormone receptor gene in two individuals with Leydig cell tumors. AB - Inactivating mutations of the luteinizing hormone receptor (LHR) gene in males induce Leydig cell agenesis or hypoplasia, while activating mutations cause testotoxicosis. Recently, it was demonstrated that a somatic heterozygous activating mutation of the LHR gene (Asp578His), limited to the tumor, was the cause of Leydig cell adenomas in three unrelated patients. We describe the molecular study of two unrelated boys with gonadotropin-independent hypersecretion of testosterone due to Leydig cell adenomas. Genomic DNA was extracted from the tumor, the adjacent normal testis tissue, and blood leukocytes. Both individuals exhibited an heterozygous missense mutation, limited only to the tumor, consisting of a guanine (G) to cytosine (C) substitution at codon 578 (GAT to CAT), turning aspartic acid into histidine. The presence of the same mutation in different ethnic groups demonstrates the existence of a mutational hot spot in the LHR gene. Indeed, this mutation occurs at the conserved aspartic acid residue at amino acid 578, where a substitution by glycine is the most common mutation observed in testotoxicosis and where a substitution by tyrosine has been linked to a more severe clinical phenotype where diffuse Leydig cell hyperplasia is found. Our results confirm the fact that somatic activating mutations of gonadotropin receptors are involved in gonadal tumorigenesis. PMID- 11857566 TI - De novo interstitial direct duplication 1(q23.1q31.1) in a fetus with Pierre Robin sequence and camptodactyly. AB - Interstitial duplications of chromosomal region 1q are rarely seen. We report the first prenatal diagnosis of pure partial trisomy 1q. The fetus was karyotyped for polyhydramnios, micrognathia, and flexion of fingers of both hands. Conventional and molecular cytogenetics showed a de novo direct duplication of the chromosomal region 1q23.1q31.1 leading to a partial trisomy 1q. At autopsy, the fetus showed Pierre Robin sequence (PRS) and camptodactyly. The main histological finding was a decreased number of motoneurons with apoptotic features in the anterior horn of the spinal cord. A literature review and our observations suggest that genetic material mapping to chromosome 1q25 could be responsible for PRS with distal arthrogryposis when this is in triple dose. PMID- 11857564 TI - Mutations in the dopamine beta-hydroxylase gene are associated with human norepinephrine deficiency. AB - Norepinephrine (NE), a key neurotransmitter of the central and peripheral nervous systems, is synthesized by dopamine beta-hydroxylase (DBH) that catalyzes oxidation of dopamine (DA) to NE. NE deficiency is a congenital disorder of unknown etiology, in which affected patients suffer profound autonomic failure. Biochemical features of the syndrome include undetectable tissue and circulating levels of NE and epinephrine, elevated levels of DA, and undetectable levels of DBH. Here, we report identification of seven novel variants including four potentially pathogenic mutations in the human DBH gene (OMIM 223360) from analysis of two unrelated patients and their families. Both patients are compound heterozygotes for variants affecting expression of DBH protein. Each carries one copy of a T-->C transversion in the splice donor site of DBH intron 1, creating a premature stop codon. In patient 1, there is a missense mutation in DBH exon 2. Patient 2 carries missense mutations in exons 1 and 6 residing in cis. We propose that NE deficiency is an autosomal recessive disorder resulting from heterogeneous molecular lesions at DBH. PMID- 11857567 TI - Patellar dislocation in Kabuki syndrome. AB - We describe four individuals (two females and two males) with Kabuki syndrome and recurrent dislocation of the patella. The age of diagnosis of patellar dislocation ranged from 11 to 23 years. One individual underwent excision of the free fragment and transfer of the tibial tuberosity with good outcome. Two required patellar brace for instability. Characteristics of individuals with the syndrome at a high risk of patellar dislocation include female, adolescence or young adulthood, joint laxity, and obesity. PMID- 11857568 TI - Exclusion of the coding sequence of the doublecortin gene as a susceptibility locus in autistic disorder. PMID- 11857569 TI - International symposium: the human genome. PMID- 11857570 TI - Germline engineering: the moral challenges. AB - Not only is the use of germline genetic engineering likely in the long run to be inevitable, there are also no convincing secular moral grounds to forbid this technology in principle. Instead, the general secular moral constraints on the use of germline genetic engineering are either procedural or without predetermined content. Nor can one develop a coherent distinction between eliminating disease and enhancing human abilities. At best, in secular morality one can establish the principle to proceed with care, though the invocation of the precautionary principle argues as much in favor of the development of germline genetic engineering as against its use. Because germline genetic engineering, by eliminating certain genetic defects, offers the prospect of decreasing human suffering and decreasing the use of prenatal diagnosis and abortion, there is an obligation, all else being equal, to change the human genome. In a post-modern world, humans face the challenge of directing their own evolution, although they share no common understanding of human destiny and purpose. Such understandings, though available within religious contexts, are not available to secular bioethics. PMID- 11857571 TI - Analysis of ten candidate genes in autism by association and linkage. AB - We studied the possible involvement of ten candidate genes in autism: proenkephalin, prodynorphin, and proprotein convertase subtilisin/kexin type 2 (opioid metabolism); tyrosine hydroxylase, dopamine receptors D2 and D5, monoamine oxidases A and B (monoaminergic system); brain-derived neurotrophic factor, and neural cell adhesion molecule (involved in neurodevelopment). Thirty eight families with two affected siblings and one family with two affected half siblings, recruited by the Paris Autism Research International Sibpair Study (PARIS), were tested using the transmission disequilibrium test and two-point affected sib-pair linkage analysis. We found no evidence for association or linkage with intragenic or linked markers. Our family sample has good power for detecting a linkage disequilibrium of 0.80. Thus, these genes are unlikely to play a major role in the families studied, but further studies in a much larger sample would be needed to highlight weaker genetic effects. PMID- 11857572 TI - Behavioral phenotypic variation in autism multiplex families: evidence for a continuous severity gradient. AB - Recent genetic investigations of autism have studied multiplex families, typically including families with multiple siblings who meet criteria for a diagnosis of autism. However, little is known about the specific behavioral characteristics of siblings with autism in these multiplex families. We investigated the behavioral phenotypic variability and similarity of 351 siblings with autism in 171 multiplex families using cluster analysis and correlations. The results of cluster analyses showed that the individuals with autism could be characterized on a severity gradient: a continuum based on severity of symptoms and impairment as measured by Autism Diagnostic Interview-Revised (ADI-R) scores, verbal-nonverbal status, and nonverbal IQ scores. Clusters based on scores from the ADI-R for the autism diagnostic criteria of the DSM-IV and nonverbal IQ scores still represented a severity gradient when the effects of verbal-nonverbal status were removed. The severity gradient was shown to be heritable, with a sib correlation of 30% or a heritability of 60%. In summary, in a sample of 171 autism multiplex families, there was no evidence of discrete behaviorally defined subgroups of affected individuals or families characterized by distinct patterns of behavioral symptoms. Rather, the clusters could be characterized along a single, heritable, continuous severity dimension. PMID- 11857573 TI - Mutation screening and transmission disequilibrium study of ATP10C in autism. AB - Autism is a complex genetic disorder. Chromosome 15 is of particular interest in this disorder, because of previous reports of individuals with autism with chromosomal abnormalities in the 15q11-q13 region. Transmission disequilibrium between polymorphisms in this region and autism has been also been reported in some, but not all studies. Recently, a novel maternally expressed gene, ATP10C, was characterized and mapped to the chromosome 15q11-q13 region, 200 kb distal to UBE3A. It encodes a putative aminophospholipid translocase likely to be involved in the asymmetric distribution of proteins in the cell membrane. Preferential maternal expression has been demonstrated in fibroblasts and brain. Because of its physical location and imprinting pattern, ATP10C was considered to be a candidate gene for chromosome 15-associated autism. In an effort to find the genes responsible for autism in this chromosomal region, 1.5 kb of the 5' flanking region, as well as the coding and splicing regions of ATP10C, were screened for sequence variants. Several polymorphic markers including five nonsynonymous SNPs were identified. To investigate transmission disequilibrium between ATP10C and autism, a family-based association study was conducted for 14 markers in 115 autism trios. No significant transmission disequilibrium was found, suggesting ATP10C is unlikely to contribute strongly to susceptibility to autism in these families. However, due to limited power to detect genes of modest effect, the possible functional role of the nonsynonymous SNPs and the functional implications of the SNPs identified from 5' flanking region and intron 2 splicing region may be evaluated in further studies. PMID- 11857574 TI - Dopamine transporter polymorphism associated with externalizing behavior problems in children. AB - Early childhood externalizing behavior is a stable and heritable pattern of aggressive and delinquent behavior that often leads to the development of serious psychiatric disorders such as conduct disorder and attention deficit hyperactivity disorder. We examined the relationship between parent reported externalizing behavior (assessed at ages 4, 7, and 9 years) and the VNTR polymorphism of the 3' untranslated region of SLC6A3 (DAT1) in a community sample of 790 children ascertained as part of our longitudinal twin and adoption studies. We applied the sibling-based methodology developed by Fulker et al. [1999: Am J Hum Genet 64:259-267] for estimating allelic association with quantitative traits, while controlling for population stratification. An extension of these methods allowed for the inclusion of monozygotic twins, dizygotic twins, siblings, and singletons. We have demonstrated that the 9-repeat variant of the DAT1 is a significant risk allele for externalizing behavior at ages 4 (P=0.001) and 7 years (P=0.02). Although the effect size was negligible at age 9 (P=0.92), a formal test of the developmental decrease in effect across the three ages was non-significant (P=0.70). PMID- 11857575 TI - Association of DRD4 in children with ADHD and comorbid conduct problems. AB - Recent family and twin study findings suggest that ADHD when comorbid with conduct problems may represent a particularly familial and heritable form of ADHD. Although several independent groups have shown association between the DRD4 7 repeat allele and ADHD, others have failed to replicate this finding. Previous TDT analyses of UK and Eire samples had also been negative. We set out to further examine the role of DRD4 but selecting a subgroup of children with ADHD and comorbid conduct problems. Families were recruited from Manchester, Ireland, Birmingham and London clinics. From these, 67 children who fulfilled diagnostic criteria for ADHD and who displayed conduct disorder symptoms were selected. TDT analysis, which had previously yielded negative results for the total sample, showed evidence of association between DRD4 and "ADHD with conduct problems" (7 repeat allele-24 transmissions, 13 non-transmissions; one-tailed P=0.05). These results provide further support for the role of DRD4 in ADHD. Furthermore, these results when considered together with family and twin study findings, suggest that those children with ADHD and comorbid conduct problems may be particularly informative for molecular genetic studies of ADHD. Further work is needed to examine these phenotype issues. PMID- 11857576 TI - Further evidence for the association between attention-deficit/hyperactivity disorder and the dopamine-beta-hydroxylase gene. AB - Attention-deficit/hyperactivity disorder (ADHD) is a very common and heterogeneous psychiatric disorder of childhood with marked inattentive, hyperactive, and impulsive symptoms. The DBH gene, the locus that encodes the enzyme dopamine-beta-hydroxylase (DbetaH), seems to be an important candidate gene for association studies, since DbetaH catalyzes the conversion of dopamine to norepinephrine. The aim of this study was to test for association between the DBH gene and ADHD in a sample of 88 Brazilian nuclear families. Haplotype relative risk (HRR) analysis of the DBH TaqI restriction site polymorphism showed a preferential transmission of the TaqI A2 allele in our whole ADHD sample (chi(2)=3.61, one-tailed P=0.03). The significant effect of the A2 allele was stronger when only families with no ADHD parental diagnosis were considered (chi(2)=5.42, one-tailed P=0.01). Our results suggest a contribution of this gene to ADHD susceptibility, partially replicating previous findings that have demonstrated an association between the DBH TaqI A2 allele and ADHD. PMID- 11857577 TI - Sex differences in shared genetic and environmental influences between conduct disorder symptoms and marijuana use in adolescents. AB - The association between conduct disorder (CD) symptoms and marijuana use (MU) was assessed in 1,480 adolescent twins participating in the National Longitudinal Study on Adolescent Health. A strong association was found between CD and MU for both males (r=.38, P<.01) and females (r=.31, P<.01). Structural equation modeling on age-corrected rank normalized scores using the program Mx indicated significant gender differences in the genetic and environmental contributions to MU. Additive genetic influences were comparable in size for males (29%) and females (24%). However, for females the influence of common environmental influences was greater (58%) than for males (36%). There was also evidence of greater common environmental influences for females, as well as greater genetic influences for males for CD, but these differences did not reach significance. In addition, for CD common environmental influences did not differ significantly from zero. Additive genetic influences accounted for 61% of the variance in CD, with the remainder being of unique environmental origin. Bivariate genetic analyses suggested a moderate genetic correlation between CD and MU (r(g)=.28) and a low unique environmental correlation (r(e)=.14). These results suggest that CD and MU share genetic influences, while environmental influences tend to be more specific to each behavior. These findings may have implications for the prevention and treatment of CD and substance use among adolescents. PMID- 11857578 TI - Association of polymorphisms of the estrogen receptor gene with anxiety-related traits in children and adolescents: a longitudinal study. AB - Anxiety problems and associated temperamental traits are multifactorial, determined by the interaction of genetic and environmental factors. Genetic effects may involve both neurotransmitters and hormones. A good candidate gene for association with anxiety-related traits is the estrogen receptor (ESRalpha). Estrogen exerts an effect on mood and behavior in humans through gene regulation on binding to estrogen receptor protein. Association between ESRalpha polymorphism and anxiety-related traits was investigated in a cohort of 680 Australian adolescents studied from 4-8 months to 15-16 years of age. Genotype frequencies were estimated for polymorphic PvuII and XbaI restriction sites in intron 1 and a microsatellite [(TA)(n)] locus 5' of ESRalpha. There was strong linkage disequilibrium between the three loci and a significant sex difference was observed in allele (for (TA)(n), PvuII) and genotype (for XbaI) frequencies. There were no significant allelic or genotypic differences in anxiety-related traits for the three loci tested. However, some significant associations were found for PvuII/(TA)(n) and XbaI/(TA)(n) two-locus genotypes and anxiety, accounting for between 1.6% and 2.8% of the total variance for anxiety in this population. The discordance in Hardy-Weinberg proportions at the XbaI locus between the sexes is an important finding, perhaps indicating a sex-specific role for ESRalpha in fetal survival. PMID- 11857580 TI - Evidence for an N-glycosylation polymorphism of arylsulfatase a predisposing to alcoholism in Koreans. AB - This study investigated the possible effect of the pseudodeficient N glycosylation polymorphism of the arylsulfatase A (ASA) gene on alcohol dependence among Koreans. Alcoholic patients (N=123) were more likely than control subjects to be heterozygous or homozygous for the ASA pseudodeficient N glycosylation site (36% of alcoholics versus 20% of controls; P<0.01). Among these 123 alcoholic patients, 42 alcoholics were heterozygous and two were homozygous for the ASA pseudodeficient N-glycosylation polymorphism. This result provides evidence that the ASA pseudodeficient N-glycosylation site allele increases the risk of alcohol dependence within a Korean population. PMID- 11857579 TI - Positive association of dopamine D2 receptor polymorphism with bipolar affective disorder in a European Multicenter Association Study of affective disorders. AB - Convincing evidence for a genetic component in the etiology of affective disorders (AD), including bipolar affective disorder (BPAD) and unipolar affective disorder (UPAD), is supported by traditional and molecular genetic studies. Most arguments lead to the complex inheritance hypothesis, suggesting that the mode of inheritance is probably not Mendelian but most likely oligogenic (or polygenic) and that the contribution of genes could be moderate or weak. The purpose of the present European multicenter study (13 centers) was to test the potential role in BPAD and UPAD of two candidate dopaminergic markers, DRD2 and DRD3, using a case-control association design. The following samples were analyzed for DRD2: 358 BPAD/358 control (C) and 133 UPAD/ 133 C subjects, and for DRD3: 325 BPAD/ 325 C and 136 UPAD/136 C subjects. Patients and controls were individually matched for sex, age ( plus minus five years) and geographical origin. Evidence for significant association between BPAD and DRD2 emerged, with an over-representation of genotype 5-5 (P=0.004) and allele 5 (P=0.002) in BPAD cases compared to controls. No association was found for DRD2 in UPAD, and for DRD3 neither in BPAD or UPAD. Our results suggest that the DRD2 microsatellite may be in linkage disequilibrium with a nearby genetic variant involved in the susceptibility to BPAD. Our large European sample allowed for replicating of some previous reported positive findings obtained in other study populations. PMID- 11857581 TI - Psychopathology, GABA, and the Rubinstein-Taybi syndrome: a review and case study. AB - An adult female with congenital Rubinstein-Taybi syndrome (RTS) and severe mental retardation is described, who presented with symptoms of severe over-activity, short attention span, mood lability, and aggressive outbursts in a cyclical pattern, suggestive of recurrent manic-like episodes. These symptoms improved significantly with divalproex (Depakote) monotherapy. Review of the existing studies showed that 10-76% of persons with RTS may be identified with similar behavioral symptoms. We postulate other persons with RTS may respond to divalproex, and there may be some relationship between the chromosome 16p13.3 deletion and gamma-aminobutyric acid (GABA) receptor or neurotransmitter abnormalities. Recent molecular genetic studies suggest a linkage of this region to bipolar mood disorder and autism, both of which were diagnosed in this patient. Further prospective study is needed of RTS persons regarding behavioral problems, comorbid psychiatric diagnoses, and treatment responses, correlated with genetic abnormalities. PMID- 11857582 TI - Search for a shared segment on chromosome 10q26 in patients with bipolar affective disorder or schizophrenia from the Faroe Islands. AB - Previous linkage studies have suggested a new locus for bipolar affective disorder and possibly also for schizophrenia on chromosome 10q26. We searched for allelic association and chromosome segment and haplotype sharing on chromosome 10q26 among distantly related patients with bipolar affective disorder or schizophrenia and controls from the relatively isolated population of the Faroe Islands by investigating 22 microsatellite markers from a 35 cM region. We used a combined approach with both assumption free tests and tests based on genealogical relationships. The 6.5 cM region between D10S1230 and D10S2322, which has been implied in previous linkage analyses, received some support. A search for segment sharing yielded empirical P-values around 0.02 among patients with bipolar affective disorder and around 0.03 for patients with schizophrenia. For both disorders combined allelic association yielded empirical P-values around 0.003 at marker D10S1723. A haplotype data mining approach supported haplotype sharing in this region. In another, more distal, 11.5 cM region between markers D10S214 and D10S505, which has received support in previous linkage studies, increased haplotype sharing in patients with bipolar affective disorder was supported by Fisher's exact test, tests based on genealogy and by haplotype data mining. Our findings yield some support for a risk gene for bipolar affective disorder and possibly also for schizophrenia. PMID- 11857583 TI - Evaluation of linkage disequilibrium between chromosome 22q11 single nucleotide polymorphisms in a large outbred population. AB - To assess the utility of linkage disequilibrium (LD) as a tool for fine-mapping disease genes in non-isolated populations, we have assessed the linkage disequilibrium strength among a series of single nucleotide polymorphisms (SNPs) in an approximate 1 Mb region of human chromosome 22q11. Nineteen random SNPs were discovered and tested across this region with an average spacing of 57 kb (range=1.4-289 kb). These 19 SNPs were genotyped in a population consisting of 444 unrelated pedigrees that were largely collected in the U.S. and U.K. Haplotypes for all pedigrees were derived from pedigree data and over 1,400 haplotypes from unrelated individuals were evaluated for linkage disequilibrium between marker alleles. In addition, linkage disequilibrium between marker alleles was also evaluated using estimated haplotypes without genealogical information (i.e., without parental genotype information). Every marker pair combination was tested for a total of 171 tests and 2x2 contingency tables were constructed to measure LD strength. In general the haplotypes derived from pedigree data provided a more conservative estimate of LD strength. Using genealogical information for estimates of D', 59% (10/17) of marker pairs less than 50 kb apart had D' values >0.30. Finally, we observed a 60 kb region with non-significant LD, which could reflect increased recombination in this region. PMID- 11857584 TI - Genetic segregation analysis of recurrent, early-onset major depression: evidence for single major locus transmission. AB - Coordinated efforts are now underway to identify susceptibility genes for unipolar major depressive disorder (MDD) and related disorders. These studies have focused on recurrent, early-onset MDD (RE-MDD), thought to be the most familial form of this disorder. The goal of this study was to conduct a complex segregation analysis of recurrent MDD and other major mood disorders aggregating in families identified by probands with RE-MDD. Eighty-one families were identified through probands over the age of 18 who met criteria for recurrent (> or =2 episodes), early-onset (< or =25 years), nonpsychotic, unipolar MDD (RE MDD) and included 407 first-degree relatives and 835 extended relatives. Psychiatric diagnoses for probands and their family members who provided blood samples were formulated from structured personal interviews, structured family history assessments, and available medical records. The remaining family members who participated and those who were deceased were evaluated through the family history method augmented by available medical records. Best-estimate diagnoses were made during a consensus conference according to established diagnostic criteria. Segregation analyses were performed using the REGD routine in S.A.G.E. release 4.0. The segregation analysis of recurrent MDD supported a sex independent Mendelian codominant model. Analysis of major mood disorders supported a sex-independent Mendelian dominant model. Interestingly, inclusion of spousal residual correlations provided better fitting models for recurrent MDD but not the broader phenotype of major mood disorders. Unlike unipolar MDD, the lifetime prevalence of bipolar I disorder in this sample of families did not exceed the reported population prevalence [Zubenko et al., 2001]. Our results suggest that a major locus contributes to the expression of recurrent MDD and possibly other major mood disorders within families identified by probands with RE-MDD. Due to the limitations of the segregation analysis model, our results cannot address whether the same major locus is segregating across families in our sample or whether multiple major loci are involved (genetic heterogeneity). The absence of aggregation of bipolar I disorder in these families strongly suggests that while the genetic determinants of unipolar and bipolar disorders may overlap, they are not identical. Our findings illustrate the advantage of employing families identified by probands with RE-MDD in studies designed to detect susceptibility loci for unipolar MDD and related disorders. PMID- 11857585 TI - Gene-based SNP genetic association study of the corticotropin-releasing hormone receptor-2 (CRHR2) in major depression. AB - An increasing amount of data suggests that affective disorders are related to dysregulation of the hypothalamic-pituitary-adrenal (HPA) axis, the stress response system. Corticotropin-releasing hormone receptor-2 (CRHR2)-deficient mice display a stress-sensitive and anxiety-like phenotype suggesting that the CRHR2 is a plausible functional candidate gene influencing the reactivity of the HPA axis and therefore the liability to develop affective disorders. In this study, a gene-based single nucleotide polymorphism (SNP) map of the corticotropin releasing hormone receptor 2 (CRHR2) was constructed containing one synonymous cSNP in exon 10, two intronic SNPs, and two SNPs in the 5' upstream regulatory region. No significant difference in allele or genotype frequency was found for four out of the five SNPs between Belgian unipolar (UP) patients and age-, gender , and ethnicity-matched controls. The cSNP did show allelic and genotypic association with borderline significance (P=0.04). However, a replication study of this cSNP in a bipolar sample of Belgian origin and a Swedish UP sample did not show significant differences in allele and genotype frequencies. PMID- 11857586 TI - Tandem pore domain K(+)-channel TASK-3 (KCNK9) and idiopathic absence epilepsies. AB - Recently, the gene coding for the tandem pore domain K(+)-channel TASK-3 (KCNK9) has been localized to the chromosomal region 8q24. Because mutations in ion channel genes have been recognized as an important factor in the etiology of abnormal neuronal excitability, TASK-3 is an interesting candidate gene for epilepsies linked to 8q24. We therefore performed a mutation analysis of the TASK 3 gene in 65 patients with childhood and juvenile absence epilepsy. Only one silent nucleotide exchange (636C/T) was detected in exon 2 of the TASK-3 coding region. No evidence for an allelic association was found between the exon 2 polymorphism and absence epilepsy. Accordingly, genetic variation of the TASK-3 coding region does not play a major role in the etiology of idiopathic absence epilepsies. PMID- 11857587 TI - Serotonin transporter promoter polymorphism, peripheral indexes of serotonin function, and personality measures in families with alcoholism. AB - A functional polymorphism in the regulatory region of the serotonin transporter gene (5-HTTLPR) is considered to be a plausible candidate gene for anxiety related personality traits and for alcoholism. Empirical support for the association between 5-HTTLPR and psychological traits has been somewhat inconsistent; however, observations of the functional dominance of the low activity s-allele over the l-allele have been more consistent. When studying the influence of particular genes on psychological traits, it seems useful also to assess more biological intermediate traits that may mediate the effects of those genes on the traits of interest. The present study examined relationships between 5-HTTLPR genotype, whole blood serotonin (5-HT) level, and platelet 5-HT binding in 150 Caucasian subjects from 50 biological families. Individuals with the s allele had lower average platelet 5-HT binding availability than those with the l/l genotype (P<0.025). Whole blood 5-HT level was not associated with 5-HTTLPR genotype. In adult men, those with the s-allele had higher mean scores on the NEO FFI personality trait of openness than did those with the l/l genotype (P=0.002). The effect was not statistically significant in women (P=0.42), although it was in the same direction. Our findings do not support an association of 5-HTTLPR genotype with alcoholism diagnosis, alcoholism subtype, or the personality trait of neuroticism. The results of this pilot study suggest that further work should examine the mediation of the genetic effects on personality traits by biochemical measures and their moderation by gender. PMID- 11857588 TI - Full genome screen for Alzheimer disease: stage II analysis. AB - We performed a two-stage genome screen to search for novel risk factors for late onset Alzheimer disease (AD). The first stage involved genotyping 292 affected sibling pairs using 237 markers spaced at approximately 20 cM intervals throughout the genome. In the second stage, we genotyped 451 affected sibling pairs (ASPs) with an additional 91 markers, in the 16 regions where the multipoint LOD score was greater than 1 in stage I. Ten regions maintained LOD scores in excess of 1 in stage II, on chromosomes 1 (peak B), 5, 6, 9 (peaks A and B), 10, 12, 19, 21, and X. Our strongest evidence for linkage was on chromosome 10, where we obtained a peak multipoint LOD score (MLS) of 3.9. The linked region on chromosome 10 spans approximately 44 cM from D10S1426 (59 cM) to D10S2327 (103 cM). To narrow this region, we tested for linkage disequilibrium with several of the stage II microsatellite markers. Of the seven markers we tested in family-based and case control samples, the only nominally positive association we found was with the 167 bp allele of marker D10S1217 (chi square=7.11, P=0.045, df=1). PMID- 11857589 TI - Search for common haplotypes on chromosome 22q in patients with schizophrenia or bipolar disorder from the Faroe Islands. AB - Chromosome 22q may harbor risk genes for schizophrenia and bipolar affective disorder. This is evidenced through genetic mapping studies, investigations of cytogenetic abnormalities, and direct examination of candidate genes. Patients with schizophrenia and bipolar affective disorder from the Faroe Islands were typed for 35 evenly distributed polymorphic markers on 22q in a search for shared risk genes in the two disorders. No single marker was strongly associated with either disease, but five two-marker segments that cluster within two regions on the chromosome have haplotypes occurring with different frequencies in patients compared to controls. Two segments were of most interest when the results of the association tests were combined with the probabilities of identity by descent of single haplotypes. For bipolar patients, the strongest evidence for a candidate region harboring a risk gene was found at a segment of at least 1.1 cM including markers D22S1161 and D22S922 (P=0.0081 in the test for association). Our results also support the a priori evidence of a susceptibility gene to schizophrenia at a segment of at least 0.45 cM including markers D22S279 and D22S276 (P=0.0075). Patients were tested for the presence of a missense mutation in the WKL1 gene encoding a putative cation channel close to segment D22S1161--D22S922, which has been associated with schizophrenia. We did not find this mutation in schizophrenic or bipolar patients or the controls from the Faroe Islands. PMID- 11857590 TI - The real problem in association studies. PMID- 11857591 TI - Clinical utility of CD23 and FMC7 antigen coexistent expression in B-cell lymphoproliferative disorder subclassification. AB - BACKGROUND: CD23 and FMC7 are normal B-cell antigens utilized during diagnostic immunophenotyping of suspected lymphoproliferative disorders. However, the diagnostic utility of coexistent antigenic expression patterns with simultaneous two-color staining and flow cytometric analysis has not been studied extensively. METHODS: Using multiparameter flow cytometry, we evaluated the expression pattern of FMC7 and CD23 in 218 cases of B-cell lymphoma from blood and bone marrow specimens. RESULTS: The CD23(+)/FMC7(-) pattern was the most common pattern in patients with chronic lymphocytic leukemia and related variants. The widest variation of patterns was found in patients with follicular cell lymphoma, large cell lymphoma, and Waldenstrom's macroglobulinemia, a lymphoplasmacytoid disorder, although most cases expressed the CD23-/FMC7(+) pattern. The CD23 and FMC7 antigen, along with the CD5 coexpression pattern, provides critical adjunctive data. These data allow accurate classification of the majority of cases, thereby providing a key aspect of a reliable diagnostic algorithm. The CD23 and FMC7 antigen expression pattern, along with selected other antigens, was predictive of subtypes in >95% of lymphoproliferative cases and narrowed the differential diagnosis in the remaining cases. CONCLUSION: The flow cytometric CD23/FMC7 expression pattern achieved by multicolor immunophenotyping facilitates accurate and reproducible classification of B-cell lymphomas and has diagnostic utility. PMID- 11857592 TI - A novel cell surface antigen, 4C8, is expressed on human eosinophils. AB - BACKGROUND: A novel monoclonal antibody, anti-4C8, reacted with human peripheral lymphocytes and monocytes but not with neutrophils. In this study, we investigated whether the 4C8 antigen is expressed on human peripheral eosinophils. METHODS: Expression of the 4C8 antigen on eosinophils was analyzed by flow cytometry and molecular analysis of the antigen was performed with eosinophils by Western blotting. RESULTS: Among human peripheral granulocytes, the 4C8 antigen was expressed on CD16-negative cells but not on CD16-positive cells. The 4C8 antigen also appeared to be expressed on eosinophils. To confirm the latter finding, eosinophils were purified from peripheral blood. On flow cytometric analysis, anti-4C8 antibody reacted with purified eosinophils. On Western blotting analysis, anti-4C8 reacted with a single band of 80 kDa in lysates from purified eosinophils. The correlation between the percentage of eosinophils determined by May-Giemsa staining and the percentage of 4C8 positive/CD16-negative cells among granulocytes was good (r = 0.91, P < 0.0001). CONCLUSIONS: Only a few cell surface antigens are available to distinguish human peripheral eosinophils from neutrophils. The novel cell surface antigen, 4C8, is a useful new marker of human eosinophils. PMID- 11857593 TI - Influence of EDTA and heparin on lipopolysaccharide binding and cell activation, evaluated at single-cell level in whole blood. AB - BACKGROUND: The use of whole blood (WB) in studying lipopolysaccharide (LPS) induced cellular activation preserves the milieu in which LPS-cell interaction occurs in vivo. However, little information is available on using such a system at a single-cell level. We evaluated LPS binding and cell activation in WB by using flow cytometry. The influence of heparin or EDTA as anticoagulants was also addressed. METHODS: Blood was obtained from healthy donors in EDTA and/or heparin tubes. Biotinylated LPS (LPSb) was used to evaluate cell binding of LPS in WB. Cells were surface stained with appropriate antibodies and LPSb was detected by adding streptavidin-allophycocyanin (APC). LPS-induced activation was evaluated by the expression of surface activation markers and by the detection of intracellular tumor necrosis factor-alpha (TNF-alpha). RESULTS: LPSb bound promptly to monocytes in EDTA- and heparin-treated blood. In EDTA-treated blood, membrane-bound LPSb decreased after 60 min of incubation, whereas it remained detectable in heparinized blood during the 6 h of incubation. LPS induced TNF alpha and enhanced the expression of HLA-DR in monocytes, as well as the expression of CD69 in T and B lymphocytes. Induction of both TNF-alpha in monocytes and CD69 in lymphocytes was more efficient in heparinized blood. CONCLUSION: Detection of membrane-bound LPSb on monocytes differed in EDTA or heparin-treated blood, and cell activation was better obtained in heparinized blood. PMID- 11857594 TI - DNA ploidy in soft tissue sarcoma: comparison of flow and image cytometry with clinical follow-up in 93 patients. AB - In soft tissue sarcoma, the prognostic importance of DNA ploidy status is limited. One possible explanation may be technical; small non-diploid stemlines will be diluted in relation to the presence of normal diploid cells and may not be detected by flow cytometry (FCM). We assessed DNA ploidy status in 93 tumors with both FCM and image cytometry (ICM). ICM may permit the exclusion of non relevant cells. The ability of the two methods to detect non-diploid stemlines was compared, as were the prognostic consequences. The patients (54 males) had a median age of 69 years. Surgical procedures were performed on all patients. None of the patients had received preoperative radiotherapy or chemotherapy. FCM and ICM were performed with standard methods. The prognostic value was assessed with univariate and multivariate analysis. In 82 of the 93 tumors, a concordant ploidy status by FCM and ICM was found. In 5 FCM type 1-2 tumors (diploid), the identification of non-diploid stemlines by ICM did not influence the metastatic rates. Increasing tumor size, histotype other than liposarcoma, increasing malignancy grade, tumor necrosis, and ICM non-diploidy were univariate prognostic factors for metastasis. In a multivariate analysis, only tumor size larger than 9 cm was a prognostic factor. In about 10% of the tumors, a discrepancy between FCM and ICM ploidy status was found, but we could not find a consistent prognostic consequence of this. Neither FCM nor ICM ploidy status was an independent prognostic factor. PMID- 11857595 TI - Androgen receptor expression and DNA content of paraffin-embedded archival human prostate tumors. AB - BACKGROUND: Androgen receptors (AR) are expressed in human prostate cells and immunohistochemistry has been used for qualitative analysis of AR expression in prostate tumor cells. Quantitative and multiparametric analysis of receptor expression could be of diagnostic and prognostic value in the management of patients on antiandrogen therapy. Multiparametric flow cytometric methods have been developed for analysis of hormone receptor expression and DNA content in nuclei isolated from formalin-fixed/paraffin-embedded human solid tumors. The present study was undertaken for analysis of AR expression and DNA content in archival human prostate tumors. METHODS: AR expression and DNA content were measured in nuclei isolated by enzyme digestion from thick sections cut from 51 paraffin-embedded human prostate tumors. AR expression in different subpopulations was studied by gated analysis. The relationship among AR activity, DNA content, and histopathological grade was analyzed. RESULTS: Distinct aneuploid populations were observed in 23% of tumors examined. AR activity was observed in all the specimens and the percentage of AR- positive nuclei in the 48 samples analyzed was <10% (n = 4), 11-50% (n = 39), and >51% (n = 5). Tumor subpopulations with aneuploid DNA content had higher AR expression (percent AR positive cells and mean log fluorescence) than the diploid subpopulations. No strong correlation was seen between AR expression and histopathological grade of the tumors. CONCLUSIONS: Flow cytometric analysis of archival prostate tumor can be used for rapid determination of aneuploid DNA content and AR expression in subpopulations of nuclei isolated from formalin-fixed/paraffin-embedded prostate tumor blocks. PMID- 11857596 TI - Precise and rapid assessment of Escherichia coli adherence to vaginal epithelial cells by flow cytometry. AB - BACKGROUND: In the pathogenesis of Escherichia coli urinary tract infections (UTIs) in women, infecting bacteria adhere to vaginal and periurethral epithelial cells prior to ascending to the bladder and causing infection. Complex interactions among specific bacterial adhesins and various host factors appear to influence adherence of E. coli to mucosal surfaces such as the urogenital epithelium. To conduct population-based studies assessing host epithelial cell determinants that influence bacterial attachment, a method of measuring bacterial adherence utilizing clinically derived epithelial cell samples is needed. METHODS: We developed and standardized an efficient, accurate, high-throughput method for analyzing the adherence of uropathogenic E. coli to clinical samples containing a large number of exfoliated vaginal epithelial cells (VEC). Three wild-type E. coli strains isolated from women with UTI (IA2 expressing pap encoded, class II fimbriae only; F24 expressing pap-encoded, class II and type 1 fimbriae; and F20, without pap-encoded or type I fimbriae) were transformed with gfpmut3, encoding green fluorescent protein, incubated with VECs, and analyzed by flow cytometry. RESULTS: Enumeration of the binding of each E. coli strain to 10,000 VECs showed reproducible, highly significant strain-dependent differences in adherence to VECs. Differential analysis of the relative contributions of type 1 pili and P fimbrial-mediated binding to the adherence phenotype was performed. It demonstrated that IA2 binding was dependent entirely on P fimbriae, whereas F24 binding was dependent on both P and type 1 fimbriae. CONCLUSIONS: This method has great potential for use in high-throughput analyses of clinically derived epithelial cell samples and will be valuable in population-based investigations of host-parasite interactions in UTI utilizing VECs collected from specific patient groups. PMID- 11857597 TI - Photothermal properties of shape memory polymer micro-actuators for treating stroke. AB - BACKGROUND AND OBJECTIVES: In this paper the photothermal engineering issues of novel shape memory polymer (SMP) microactuators for treating stroke are presented. The engineering issues for using lasers to heat and subsequently actuate these SMP devices are presented in order to provide design criteria and guidelines for intravascular, laser activated SMP devices. MATERIALS AND METHODS: A total of three devices will be presented: two interventional ischemic stroke devices (coil and umbrella) and one device for releasing embolic coils (microgripper). The optical properties of SMP, methods for coupling laser light into SMP, heating distributions in the SMP devices, and the impact of operating the thermally activated material in a blood vessel are presented. RESULTS: Actuating the devices requires device temperatures in the range of 65-85 degrees C. Attaining these temperatures under flow conditions requires critical engineering of the SMP optical properties, optical coupling into the SMP, and device geometries. CONCLUSION: Laser-activated SMP devices are a unique combination of laser-tissue and biomaterial technologies. Successful deployment of the microactuator requires well-engineered coupling of the light from the diffusing fiber through the blood into the SMP. PMID- 11857598 TI - Comparison of 5-aminolaevulinic acid and porphyrin photosensitization for photodynamic therapy of malignant bronchial stenosis: a clinical pilot study. AB - BACKGROUND AND OBJECTIVES: Photosan, a mixture of porphyrin oligomers as sensitizer for photodynamic therapy (PDT), carry the risk of prolonged photosensitivity of the skin. New sensitizer such as 5-aminolaevulinic acid (ALA) with low rates of skin phototoxicity appear to be promising alternatives. The aim of this study was to evaluate the efficacy of ALA compared to Photosan for PDT in malignant tracheo-bronchial stenosis. Reduction of tumor stenosis, increase in quality of life, and phototoxicity were considered as primary objectives. Improvement in clinical symptoms due to reduction of tumor stenosis, for example hemotysis, dyspnea, and poststenotic pneumonia were considered as secondary objectives. PATIENTS AND METHODS: After diagnostic work-up, photosensitization was done in 16 patients with ALA (60 mg/kg BW, oral, 6-8 hours prior to PDT) and in 24 patients with Photosan (2 mg/kg BW, i.v., 48 hours before PDT). The light dose was calculated as 100 J/cm(2) tumor length. Light at 630 nm was applied using a pumped dye laser. In both groups, additional hyperbaric oxygenation was applied at a level of 2 absolute atmospheric pressure. RESULTS: Stenosis diameter and Karnofsky performance status showed a significant improvement in favor of the Photosan-group, P = 0.00073 and 0.00015, respectively. In both groups no sunburn occurred due to phototoxicity of the sensitizer. CONCLUSION: Despite the limitations of a non-randomized study, photosensitization with Photosan seems to be more effective in PDT of malignant tracheo-bronchial stenosis compared to ALA. PMID- 11857599 TI - Photo-induced cytomorphologic changes in an advanced cancer phase I clinical trial. AB - BACKGROUND AND OBJECTIVES: The aim of this study was to investigate whether the application of an Infrared Pulsed Laser Device (IPLD) photo-induced significant cytomorphologic changes during the monitoring of advanced cancer patients participating in a phase I clinical trial. MATERIALS AND METHODS: Patients were irradiated with an IPLD (904 nm pulsed at 3 MHz) under a one-dose, one-schedule, and one-procedure design. Total daily dose consisted of a Radiant Exposure of 4.5x10(5) J/m(2). Thirty-one tissue samples from eleven patients with progressive solid neoplastic diseases (TNM IV, UICC) were obtained at three intervals: Time 0 (15-90 days pre-treatment, n=11); Time I (2-5 months post-treatment; n=11); Time II (6-12 months post-treatment, n=09). Three blinded pathologists evaluated samples; scores were determined by consensus. Data were evaluated by using the Wilcoxon matched-pairs signed-rank test and Spearman rank correlation coefficient. The level of statistical significance was alpha=0.05. RESULTS: Increased apoptosis (Time I, P<0.003; Time II, P<0.007), necrosis (Time I, NS; Time II, P<0.01), cytoplasmic vacuoles (Time I, P<0.03; Time II, P<0.02), and nuclear vacuoles (Time I, NS; Time II, P<0.01), reduced cell size (Time I, P<0.007; Time II, P<0.01) and intercellular adhesion (Time I, P<0.01; Time II, P<0.02) were present in neoplastic cells after IPLD treatment. No apparent changes were noted in non-neoplastic cells. The Spearman rank correlation coefficient between apoptosis, necrosis, nuclear vacuoles, cytoplasmatic vacuoles, intercellular adhesion, and cell size was positive and highly significant (P<0.006). CONCLUSIONS: Although further research is necessary, our preliminary results support the novel possibility that the IPLD photo-induces chaotic dynamics that modulate complex physiologically reparative bioeffects. PMID- 11857600 TI - Photodynamic therapy using Verteporfin (benzoporphyrin derivative monoacid ring A, BPD-MA) and 630 nm laser light in canine esophagus. AB - BACKGROUND AND OBJECTIVE: Verteporfin is a new photosensitizer with short-term skin photosensitivity. The objective of this preclinical study was to find the light dose that effectively ablates canine esophageal mucosa when delivered 30 minutes after Verteporfin injection. STUDY DESIGN/MATERIALS AND METHODS: Verteporfin was administered intravenously (0.75 mg/kg). 630 nm light from KTP/Dye laser was delivered using an esophageal Photodynamic therapy (PDT) balloon. In Phase I study, animals were treated 30 minutes after drug injection using 40, 60, and 80 J/cm to find the desired light dose. Using results from phase I and application of reciprocity principle (light dose vs. plasma concentration of drug), additional light doses were calculated for delivery at other times. In phase II, animals were treated at 15, 60, and 120 minutes, using the calculated light doses of 60, 145, and 200 J/cm, respectively. Animals were followed for 2 days to 4 weeks. RESULTS: In Phase I, 80 J/cm at 30 minutes induced total mucosal ablation. In Phase II, light doses of 60, 145, and 200 J/cm induced similar mucosal injuries when delivered at 15, 60, and 120 minutes, respectively. CONCLUSIONS: Effective mucosal ablation in canine esophagus was achieved using Verteporfin and 630 nm light doses of 60, 80, 145, and 200 J/cm when delivered at 15, 30, 60, and 120 minutes after the drug injection, respectively. PMID- 11857601 TI - Comparison of laser induced effects on hyperplastic inferior nasal turbinates by means of scanning electron microscopy. AB - BACKGROUND AND OBJECTIVE: Various laser systems have been used for volume reduction of hyperplastic inferior nasal turbinates. Many studies have thus reported about clinical experiences concerning short and long term results. Although there are also some clinical investigations on histological changes after laser treatment, there is a lack of examinations by means of scanning electron microscopy (SEM) in order to especially investigate laser induced superficial tissue effects (ablation, carbonization, and coagulation). MATERIALS AND METHODS: Directly following operative excision, three hyperplastic nasal turbinates were treated by laser light of five common medical laser systems and their corresponding wavelengths, using identical laser parameters (irradiance, laser spot diameter) and a reproducible application procedure. After passing through an ethanol dehydration process, the thermal effects on the tissue (carbonization, coagulation, and ablation) were investigated and photographed via SEM. RESULTS: Depending on the laser wavelength used, tissue responses which could be observed by SEM greatly differ with respect to ablation, coagulation, and carbonization zones. While the CO(2) laser showed a precise cutting of tissue via ablation with only slight thermal effects on the adjoining tissue, Argon-ion , Nd:YAG-, and diode-lasers provided for an intense destruction of both the surrounding, superficial respiratory epithelium, and the tissue in the depth of the turbinates, resulting from the development of rather large carbonization and coagulation zones. Ho:YAG-laser light induced precise vaporization in addition to remarkable superficial coagulation of the tissue, which was of smaller extent than after treatment with Nd:YAG- and diode-laser, but comparable to the Argon ion-laser. CONCLUSIONS: In the course of this study SEM proved to be especially suitable for the examination of tissue changes on the surface of laser treated turbinates (i.g., destruction of respiratory epithelium). According to our results, Argon-ion-, Nd:YAG-, and diode-lasers seem to be useful for an effective and bloodless reduction of hyperplastic turbinates. However, broad coagulation zones may lead to consecutive complications and side effects. Due to its good ablation and superficial coagulation capabilities, Ho:YAG-laser light also seems to be usable for an effective turbinate reduction, but its low thermal effects in the depth of the tissue may lead to bleedings during treatment. Laser light of the CO(2) laser showed the lowest thermal tissue effects of the lasers investigated and therefore holds the risk of bleedings during treatment and the need of more than one therapeutic session to get a sufficient result. This investigation revealed that physicians, who are performing endonasal laser treatment, must be aware of the varying tissue effects of different medical laser systems to get a more aimed effect and to avoid intra- and post-operative complications (e.g., Rhinitis sicca, bone sequestration of the turbinate, bleedings, ...). PMID- 11857602 TI - Laser soft palate "stiffening": an alternative to uvulopalatopharyngoplasty. AB - BACKGROUND AND OBJECTIVE: Habitual snoring is best treated surgically, with uvulopalatophayngoplasty, but the standard "cutting" procedure poses potential morbidity. In this study, a new approach, stiffening of the soft palate with a low energy laser, was studied as a less invasive alternative. Study Design In an in vitro study, five fresh palates of canines were used to see acute thermal effect on the palatal tissue after laser treatment. The same laser irradiation was conducted in three living canines to observe a delayed response of the palate to the laser. MATERIALS AND METHODS: With the use of a non-contact 1.44 microm Nd:YAG laser at 0.1 J and 20 Hz, the mucosa on the marginal area of the soft palate was irradiated for 2-3 minutes. Following Outcome were Measured Acute shrinkage of the soft palate and thermal effect on the mucosa, and delayed stiffening and elevation of the palate after laser irradiation. RESULTS: There was immediate shrinkage of the palate of about 3.0 mm in the in vitro study. In the in vivo study, a delayed palatal stiffening with 6.0-7.0 mm elevated palatal arch was found at 5 weeks. There was no morbidity after the treatment. CONCLUSION: Laser stiffening of soft palate is simple, safe and effective for reduction of length and fluttering of the soft palate in the canine model. Clinical studies are warranted to evaluate its efficacy as an office treatment for snoring. PMID- 11857603 TI - Laser treatment of congenital facial port-wine stains: long-term efficacy and complication in Chinese patients. AB - BACKGROUND AND OBJECTIVE: Pulsed dye laser (585 nm, 450 micros) and variable pulse width frequency doubled Nd:YAG 532 nm laser have been proven to be effective in treating vascular lesions in Caucasians especially in facial port wine stain. Their roles in dark-skinned Asian have not been determined. The objective of this study was to assess the long-term outcome and complication of laser treatment of congenital facial port-wine stain in Chinese patients retrospectively. STUDY DESIGN/MATERIALS AND METHODS: Files of all patients with port-wine stains who had received laser treatment in a major regional laser center in a 5-year period were reviewed. Chinese patients with congenital facial port-wine stains were recruited for the study. They were called back for questionnaire assessment of their degree of clearance and clinical examination for treatment complication. Detail demographics, result of questionnaire and clinical examination were entered into a database and statistical analyses were conducted where applicable. RESULTS: One hundred and seven patients with congenital facial port-wine stain were recruited for the study. Thirty-six patients had received PDL treatment, 40 patients had VP532 treatment and 31 patients had been treated by both laser systems. A total of 665 treatment sessions had been performed with an average of 6.1 +/- 2.8 sessions (range 3-12). They had been followed-up from 2 to 5 years with a mean period of 3.4 +/- 1.1 years. Over 60% of patients had more than 25% of clearing and the majority of patients (41.1%) had 25-50% of clearing. Less than one-quarter of patients (23%) experienced a clearing more than 50% and no patient had complete clearing. There was no significant difference in the subjective assessment of clearing between patients that had different forms of laser treatments. No patient had recurrence of pigmentation after treatment. Fourteen percent of patients experienced complications and the majority of them had pigmentary changes. Sex, age, number of treatment session, and type of laser machine used were not related to a higher complication rate using the multiple regression analysis. CONCLUSIONS: Chinese patients were less responsive to laser treatment with higher complication rate. In addition, more treatment sessions were required to achieve a maximum clearing. However, most of our patients were satisfied with laser treatment (Patient Satisfaction Score=7.25). Dark-skinned patients should not be excluded from laser therapy provided that treatment expectations and risks are fully discussed. PMID- 11857604 TI - Ex vivo evaluation of human fetal membrane closure. AB - BACKGROUND AND OBJECTIVE: This study compares stress and leak point pressure of human fetal membranes following closure with suture, laser welding, or a new tissue sealant, SynthaSeal. STUDY DESIGN/MATERIALS AND METHODS: Samples of fetal membrane were bisected and repaired with suture, laser welding with albumin solders, or a tissue sealant. Representative tissue samples were assessed by histology. Anastomotic strength was evaluated by tensiometry. Additionally, leak pressure was measured after membrane samples were placed onto an infusion pump/pressure transducer, incised, and repaired as above. RESULTS: Membrane stress was strongest (P<0.001) for the tissue sealant group. Sutured samples were stronger than laser-welded samples. Membrane leak pressure was greatest for the tissue sealant group. The suture group demonstrated a higher leak pressure than the laser-welded group. CONCLUSIONS: Human fetal membranes were not suited for repair with laser welding under the conditions tested. A new tissue sealant, SynthaSeal, may provide a simple, effective method for membrane closure following fetoscopy, amniocentesis, and open fetal surgery. PMID- 11857605 TI - Measurement of the surface temperature of the cornea during ArF excimer laser ablation by thermal radiometry with a 15-nanosecond time response. AB - BACKGROUND AND OBJECTIVE: The purpose of this work was to develop a temperature measurement system with a nanosecond time response to monitor the transient temperature of the corneal surface during laser refractive surgery. MATERIALS AND METHODS: Thermal radiation from the surface of the porcine cornea during ArF excimer laser irradiation was measured using a photovoltaic HgCdTe detector with a response bandwidth of 150 MHz. RESULTS: Maximum thermal radiation occurred at 31 +/- 4 nanoseconds, which was longer than the time response of the measurement system. The temperature derived from the detected signal reached over 100 degrees C at a fluence of 80 mJ/cm(2), which was the ablation threshold, and reached 240 degrees C at a fluence of 180 mJ/cm(2). CONCLUSION: The present system of temperature measurement with a time response of 15.7 nanoseconds revealed that the transient surface temperature of the cornea during ablation is much higher than that previously reported. PMID- 11857606 TI - Effect of gallium arsenide diode laser on human periodontal disease: a microbiological and clinical study. AB - BACKGROUND AND OBJECTIVE: The present study is aimed to describe short-term results on selected microbiological and clinical parameters obtained by treatment with soft laser in conjunction with methylene blue and/or mechanical subgingival debridement in human periodontal disease. STUDY DESIGN/MATERIALS AND METHODS: Ten patients, in whom each dental quadrant was randomly designated to receive one of four types of treatment procedures, were included in the study. Groups of quadrants received: scaling/root planing (SRP); laser application (L); SRP combined with L (SRP/L); oral hygiene instructions (OHI). Four single rooted teeth (one in each quadrant), having an interproximal site with a probing depth of 4 mm mesio-buccally, were selected in each patient. The selected teeth were first assessed for microbiological (one site/tooth) and then for clinical variables (six sites/tooth). Supragingival irrigation with methylene blue was performed prior to laser application. The microbiological (proportions of obligate anaerobes) and clinical measurements (plaque and gingival indices, bleeding on probing, probing pocket depth) were evaluated over a period of 32 days. RESULTS: Only the SRP/L and SRP groups provided significant reductions in the proportions of obligate anaerobes before and after treatments with no significant differences in between. Parallel to the microbiological changes, both SRP/L and SRP resulted in similar clinical improvements, whereas L alone revealed a limited effect similar to OHI. CONCLUSION: Within the limits of this study, methylene blue/soft laser therapy provided no additional microbiological and clinical benefits over conventional mechanical debridement. PMID- 11857607 TI - A laser pumped Nd(3+)-doped YAG fiber-optic thermal tip for laser thermotherapy. AB - BACKGROUND AND OBJECTIVE: Photo-thermal converters have been successfully used in laser surgery; however, most of the currently used converters are likely to be damaged in the operations due to various problems, preventing them from being efficiently and repeatedly used in laser thermotherapy. Aiming to solve these problems, we report a new type of fiber-optic photo-thermal converter with higher stability. STUDY DESIGN/MATERIALS AND METHODS: A laser-pumped highly Nd(3+)-doped Y(3)Al(5)O(12) (YAG) crystal fiber tip had been fabricated for photo-thermal conversion by means of phonon relaxation. The 15at.% neodymium-doped near cylindrical YAG crystal fiber tip, with maximum diameter of 1.1 mm and length of 2.7 mm, was fabricated on a 1 mm-thick 42 mm-long pure YAG single crystal fiber by laser heated pedestal growth (LHPG) method. RESULTS AND CONCLUSION: Pumping by a Ti:Sapphire laser with central wavelength of 805 nm, the tip generated a temperature rise of 15 degrees C in egg white with pumping power of 710 mW, the tip temperature increased approximately linearly with the increased pumping power. The distribution of the temperature in two media had also been measured. Experimental results indicated that the tip was promising for laser thermotherapy applications. PMID- 11857608 TI - Congenital lymphedema presenting with increased nuchal translucency at 13 weeks of gestation. AB - Congenital lymphedema is an autosomal dominant condition characterized by chronic tissue swelling caused by deficient lymphatic drainage due to hypoplastic/aplastic lymphatic vessels and usually affecting the lower limbs. The locus of the gene has been identified in the long arm of chromosome 15. We report one case of congenital lymphedema presenting with increased nuchal translucency at 13 weeks of gestation. PMID- 11857609 TI - Congenital nephrotic syndrome presenting with increased nuchal translucency in the first trimester. AB - Congenital nephrotic syndrome of the Finnish type (CNF) and diffuse mesangial sclerosis (DMS) are rare causes of renal failure in infants. We report two cases, one of each condition, presenting with increased nuchal translucency at the 11-14 week scan, and review the literature. PMID- 11857610 TI - The management of fetal alloimmune thrombocytopenia. PMID- 11857611 TI - Thermocoagulation of fetal sacrococcygeal teratoma. AB - Fetal sacrococcygeal teratoma can lead to a high output cardiac failure resulting in hydrops fetalis. One of the prenatal therapeutic options is to occlude the feeding vessels by radiofrequency ablation. We present a case of fetal sacrococcygeal teratoma diagnosed at 13 weeks of gestation. The tumour increased in size more than 100 fold over 5 weeks causing polyhydramnios and cardiac and placental enlargement. Thermocoagulation was performed at 18 weeks' gestation by passing an insulated electric wire through an 18 gauge needle placed close to the feeding vessels of the tumour at its neck. Blood supply to the tumour was successfully reduced. However, fetal death was diagnosed 2 days after the procedure. We speculate that it may be safer to limit the extent of coagulation in one attempt but to repeat the procedure at a later stage when necessary. PMID- 11857612 TI - Prenatal diagnosis of a familial complex chromosomal rearrangement involving chromosomes 5, 10, 16 and 18. AB - We report one case of a familial complex chromosomal rearrangement (CCR) involving four different chromosomes 5, 10, 16 and 18. The CCR was detected prenatally at 20 weeks' gestation because of advanced maternal age and history of recurrent miscarriages. Cytogenetic analysis of cultured amniotic fluid cells with GTG banding showed a 46,XX,t(5;16;10;18)(q13;q22;q11.2;q21) karyotype. Parental cytogenetic study revealed that the mother has the same CCR. RBG banding, high resolution banding and fluorescence in situ hybridization (FISH) were used to characterize further and confirm the conventional banding data. No physical abnormalities were shown in the targeted fetal ultrasonography examination. The parents decided to continue the pregnancy. The child is now 2 years old and has neither congenital anomalies nor evidence of delayed psychomotor development. The fetal targeted ultrasound and FISH analysis helped us reassure fetal status. PMID- 11857613 TI - Prenatal diagnosis of 45,X and 45,X mosaicism: the need for thorough cytogenetic and clinical evaluations. AB - OBJECTIVES: Mosaicism involving a 45,X cell line is relatively common in prenatal diagnosis. In prenatally diagnosed cases, the prognosis of non-mosaic 45,X and 45,X/46,XY mosaicism are different. Therefore, accurate identification of a cell line containing Y chromosome is critical for genetic counseling and postnatal management. METHODS: We investigated the ultrasound findings and outcomes of pregnancies with a 45,X cell line identified during mid-trimester cytogenetic analysis. RESULTS: A total of 105 cases were found to have a 45,X cell line by standard cytogenetic analysis. Seventy-four cases were found to have non-mosaic 45,X at initial diagnosis. Of these 74 cases, 68 had abnormal ultrasound findings that were characteristic of Turner syndrome. Of the six cases with normal ultrasound findings, ultrasound examination was normal with male genitalia identified in three cases. Thorough cytogenetic and fluorescent in situ hybridization (FISH) analysis identified Y chromosome material in all three cases, one with a dicentric Y;14 chromosome and the other two cases with a marker chromosome containing Sex-determining Region (SRY) material in a small portion of the cells. In contrast, in 31 cases with a mosaic 45,X karyotype, ultrasound abnormality was identified only in one case. CONCLUSIONS: The present data suggest the need for follow-up ultrasound examination and thorough cytogenetic and molecular analysis for Y chromosome material in 45,X cases with normal ultrasound findings. PMID- 11857614 TI - Fetal, neonatal cord, and maternal plasma concentrations of angiotensin converting enzyme (ACE). AB - OBJECTIVES: Angiotensin converting enzyme (ACE), a component of the renin angiotensin system (RAS), catalyses the degradation of angiotensin I to angiotensin II. It was the aim of the present study to measure ACE activity in human fetal blood and to determine its changes with advancing gestational age. METHODS: Fetal blood was sampled by cordocentesis from six control fetuses and six fetuses with Rh isoimmunisation. Cord blood was sampled from six preterm neonates, 15 neonates after spontaneous delivery at term and six neonates at term after caesarean section. In addition, maternal ACE values were determined. ACE activity was measured using the miniaturised fluorimetric method. RESULTS: In normal fetuses (13.31+/-1.41 nmol HL/min/ml) and fetuses with Rh isoimmunisation (13.08+/-2.00 nmol HL/min/ml, p<0.05). Neonatal cord blood of preterm newborns (10.43+/-0.69 nmol HL/min/ml) and term newborns (8.99+/-0.49 nmol HL/min/ml) showed a significantly decreased ACE activity compared to the fetal controls. CONCLUSION: We conclude that the high fetal ACE activity and the stringent regulation with advancing gestational age indicate the physiological importance of the enzyme during prenatal development. PMID- 11857615 TI - The development of the fetal thymus: an in utero sonographic evaluation. AB - OBJECTIVES: To establish a nomogram for fetal thymus size during gestation. METHODS: The study is a prospective, cross sectional evaluation of 403 male and female fetuses between 14 and 38 weeks of normal singleton pregnancies. Measurements of fetal thymus size were performed by high resolution transvaginal ultrasonography between 14 and 17 weeks' gestation, and by transabdominal ultrasonography after 18 weeks' gestation. RESULTS: Adequate thymus size measurements were obtained in all 403 fetuses. Thymus size as a function of gestational age was expressed by the regression equation: (square root) thymus size (mm)= -39.39+4.41 x gestational age (weeks). The correlation coefficient, r=0.965, was found to be highly statistically significant (p<0.0001). The normal mean and the 90% prediction limits were defined. CONCLUSION: The present data offer the normal range of fetal thymus size from early stages of gestation that may allow intrauterine assessment of its development. It may be helpful in the prenatal diagnosis of thymic pathologies. PMID- 11857616 TI - Early second trimester prenatal diagnosis of Neu-Laxova syndrome. AB - Neu-Laxova is a rare, uniformly lethal, autosomal recessive condition with characteristic limb posturing, facial dysmorphic features, and central nervous system abnormalities. Forty-two cases of Neu-Laxova syndrome have been reported, with only four of these diagnosed prenatally. Three of the four cases were diagnosed at or after 32 weeks' gestation. The fourth case was diagnosed at 22 weeks' gestation in a patient who was followed with serial ultrasound studies due to having a prior affected child. At 19 weeks' gestation, we present the earliest reported prenatal diagnosis of Neu-Laxova syndrome in a primigravida with a non informative family history. PMID- 11857617 TI - Prenatal diagnosis of a lethal form of Netherton syndrome by SPINK5 mutation analysis. AB - Netherton syndrome (NS) is a severe autosomal recessive ichthyosis with no specific treatment or prenatal diagnosis available at present. The recent identification of SPINK5, which encodes a serine protease inhibitor, as the defective gene enables DNA based prenatal diagnosis to be carried out. Here we report the first direct molecular prenatal diagnosis of a lethal form due to a recurrent SPINK5 mutation in three consanguineous Turkish families. XmnI restriction enzyme digestion and DNA sequencing demonstrated that each deceased affected child was homozygous for mutation 153delT inherited from each parent. Analysis of fetal DNA from amniotic fluid cells in Family 1 and from a chorionic villus sampling in Family 3 showed that the fetus was heterozygous for 153delT in both cases. The pregnancies were carried to term and the newborns were unaffected. In Family 2, fetal DNA analysis from chorionic villus biopsy showed in a first pregnancy that the fetus was homozygous for 153delT. The pregnancy was terminated at 13 weeks and DNA analysis of fetal keratinocytes confirmed the prenatal prediction. In a second pregnancy in Family 2, fetal DNA analysis showed heterozygosity for 153delT, and the pregnancy was continued. Direct SPINK5 mutation analysis in families at risk for NS represents the first early, rapid and reliable method for prenatal diagnosis of this life threatening form of ichthyosis. PMID- 11857618 TI - Prenatal diagnosis of partial tetrasomy 14: a case study. AB - Prenatal specimens were received from a fetus with abnormalities noted on ultrasound. A supernumerary marker chromosome (SMC) was detected: 47,XY,+mar. Fluorescence in situ hybridisation (FISH) further classified this to be partial tetrasomy for chromosome 14. We compare this finding with other cases of SMC (14) and further classify phenotype with karyotype. PMID- 11857619 TI - Prenatal diagnosis of ventricular aneurysm: a report of two cases and a review. AB - Ventricular aneurysms have rarely been reported prenatally. Their prognosis is variable depending on factors such as early detection, their relative size in comparison to the ventricular cavity, growth on follow-up, and signs of cardiac failure. In view of the fact that it may be useful to report on additional cases in order to make available further information on aetiology, prognosis and neonatal management, we hereby report on two cases of ventricular aneurysm with good mid-term prognosis. One case of apical aneurysm of the left ventricle was associated with a muscular ventricular septal defect, the features of which are compatible with a fetal myocardial infarction and ventricular septal rupture in absence of coronary artery anomalies as demonstrated by catheterisation. Another case of sub-tricuspidal aneurysm of the right ventricle associated with mitral prolapse appears to be a component of diffuse connective dysplasia. Despite the early gestational age at diagnosis, the large size of the aneurysm and the associated defect, both the infants were asymptomatic in infancy prompting a conservative management. PMID- 11857620 TI - A second case of intrauterine growth retardation and primary hypospadias associated with a trisomy 22 placenta but with biparental inheritance of chromosome 22 in the fetus. AB - We report a case of severe intrauterine growth retardation (IUGR) and hypospadias in association with trisomy 22 diagnosed following chorionic villus sampling (CVS). Subsequent analysis of amniotic fluid cultures showed a normal male karyotype, 46,XY. As a previous case had been reported with similar abnormalities, in association with maternal uniparental disomy (UPD) 22, molecular studies were also performed. Microsatellite marker studies showed biparental inheritance. Follow-up studies after delivery showed a normal cell line in lymphocytes with the trisomy appearing to be confined to the placenta. The present case concurs with other earlier reports that maternal UPD for chromosome 22 has no impact on the phenotype. The features seen in the fetus are most likely the result of placental dysfunction due to trisomy, tissue-specific mosaicism and/or the effects of local growth restriction. PMID- 11857621 TI - The importance of investigating for uniparental disomy in prenatally identified balanced acrocentric rearrangements. AB - We report the finding of paternal isodisomy for chromosome 14 in a fetus found to have a der(14;14)(q10;q10) by amniocentesis. The pregnancy was complicated by severe polyhydramnios and elevated amniotic fluid alpha-fetoprotein (AFP). The infant showed features consistent with paternal uniparental disomy (UPD) including postnatal growth retardation, poor respiratory function, feeding difficulties, and evidence of hypertrophic cardiomyopathy. The present case, in addition to other reported cases of UPD involving balanced acrocentric rearrangements, supports testing for UPD in prenatally detected Robertsonian translocations and isochromosomes. PMID- 11857622 TI - Fetal diaphragmatic hernia and upper limb anomalies suggest Brachmann-de Lange syndrome. AB - We describe two independent cases of Brachmann-de Lange syndrome (BDLS) in which second trimester fetal sonographic studies showed the presence of a diaphragmatic hernia and upper limb anomalies. In both cases the karyotypes were normal. Intrauterine growth restriction (IUGR) developed in the third trimester. Postnatal and postmortem physical examinations demonstrated typical physical findings associated with BDLS. The prenatal diagnosis of diaphragmatic hernia with associated anomalies should prompt consideration of an underlying genetic etiology. PMID- 11857623 TI - Testing normality of fetal DNA concentration in maternal plasma at 10-12 completed weeks' gestation: a preliminary approach to a new marker for genetic screening. AB - OBJECTIVES: To test the distribution of fetal DNA in maternal plasma expressed as gen/eq in a population of normal pregnancies. METHODS: Peripheral blood samples were obtained from 63 women (85% > or =35 years of age at delivery) bearing a euploid male fetus. Each patient underwent chorionic villus sampling (CVS) for karyotype analysis and/or beta thalassemia screening. Ultrasound scanning was used to determine gestational age. At 10-12 weeks' gestation, a peripheral blood sample was collected followed by CVS. To detect the Y chromosome specific sequences (SRY) quantitative polymerase chain reaction (PCR) analysis was used. Normal distribution of the data was tested by means of the Kolmogorov-Smirnov (KS) test. A Symmetry test (reliability p>0.05) was used to evaluate the reliability of the median. RESULTS: Only after natural logarithmic transformation did the data display a normal distribution. The median value of fetal DNA was 23.3 gen/eq (range 2.08-195), interquartile range 18.57-45.4. A Pearson test showed a significant correlation between gestational age and fetal DNA concentration (r=0.25, p=0.045). CONCLUSION: The present finding is a preliminary step towards a possible integration of fetal DNA with other variables (biochemical and/or ultrasound). It may serve to improve the discrimination of the screening for genetic diseases in the first trimester. Because of the relatively high dispersion, adjustments for possible covariates would appear to be necessary in further studies. PMID- 11857624 TI - Usefulness of automated chromatography for rapid fetal blood analysis for second trimester prenatal diagnosis of beta-thalassemia. AB - Prenatal diagnosis of beta-thalassemia is now ideally done in the first trimester of pregnancy by chorionic villus tissue DNA analysis. Nevertheless, fetal blood analysis in the second trimester is required either when the mutation in both parents cannot be characterised or when the couple comes late for investigations. We evaluated the usefulness of analysis of fetal blood on the Biorad Variant Hemoglobin Testing System using the beta-thalassemia short programme in comparison with the conventional globin biosynthesis in 58 pregnancies. The beta/alpha biosynthesis ratios in 13 homozygous fetuses ranged from 0 to 0.03 and the adult hemoglobin (HbA) levels by automated chromatography varied from 0% to 0.4%. The normal or heterozygous fetuses had beta/alpha ratios of >0.04 and HbA levels ranging from 2.1% to 10.6%. In 17 fetuses we also correlated the beta gene mutations with the predicted genotypes using automated high-performance liquid chromatography (HPLC). Follow-up of 18 unaffected fetuses using the Variant System at birth showed a significant increase in HbA levels. PMID- 11857625 TI - Re: embryonic and fetal globins are expressed in adult erythroid progenitor cells and in erythroid cell cultures (Lau et al., Prenat Diagn 2001; 21: 529-539). PMID- 11857627 TI - Sonographic visualization of fetal fingers and toes at 10 weeks of gestation. PMID- 11857628 TI - Additional studies warranted to confirm monosomy 21. PMID- 11857629 TI - Correlation between maternal serum PAPP-A and inhibin. PMID- 11857630 TI - Isolated mild fetal cerebral ventriculomegaly. PMID- 11857631 TI - In utero diagnosis of fetal gastroschisis with eviscerated gastric perforation. PMID- 11857633 TI - Fetal cervico-mediastinal cystic hygroma associated with maternal serum screening positive for Down syndrome. PMID- 11857632 TI - Minimal volume of amniotic fluid for reliable prenatal cytogenetic diagnosis. PMID- 11857634 TI - Current awareness. PMID- 11857635 TI - The cryptophycins: their synthesis and anticancer activity. AB - The cryptophycins are a unique family of 16-membered macrolide antimitotic agents isolated from the cyanobacteria Nostoc sp. Their molecular target is tubulin protein wherein they are the most potent known stabilizers of microtubule dynamics and depolymerize microtubules at higher concentrations. They also deactivate the Bcl2 protein and produce apoptotic response much more quickly and at considerably lower concentrations than clinically utilized compounds. The presence of several amide and ester linkages within the cryptophycin core provides access to very convergent total synthetic approaches. Likewise, the modularity of the structure renders their synthesis amenable to structure activity studies in several regions of the molecule. The in vivo hydrolytic instability of the C5 ester was a key obstacle to the successful identification of a clinical candidate. This problem was ameliorated by increased substitution at C6 as in the presence of gem-dimethyl substitution in the clinical candidate, cryptophycin-52. PMID- 11857636 TI - Natural products as probes of cell biology: 20 years of didemnin research. AB - The discovery of the didemnin family of marine depsipeptides launched an exciting and intriguing chapter in natural product chemistry. The unusual structure of the didemnin congeners has led to several total syntheses by research groups from around the world. The impressive in vitro and in vivo biological activities of the didemnins resulted in the first human clinical trials in the U.S. of a marine natural product against cancer, and additional clinical trials of a second generation didemnin, dehydrodidemnin B (aplidine), are underway. As we mark the 20-year anniversary of the discovery of the didemnins, this class of natural products continues to stimulate active research in fields ranging from synthetic and medicinal chemistry to clinical oncology and cell biology. While some progress was made in dissecting the molecular mechanism of action and in establishing structure-activity relationships, there are still more questions than answers. This review covers the recent didemnin literature, highlighting the work directed towards understanding how this group of natural products interact with fundamental processes such as cell proliferation, protein biosynthesis, and apoptosis. The didemnin field illustrates how natural product chemistry may be used as a critical tool for the study of cell biology. PMID- 11857637 TI - Inhibition of LFA-1/ICAM-1 and VLA-4/VCAM-1 as a therapeutic approach to inflammation and autoimmune diseases. AB - This review focuses on providing insights into the structural basis and clinical relevance of LFA-1 and VLA-4 inhibition by peptides and small molecules as adhesion-based therapeutic strategies for inflammation and autoimmune diseases. Interactions of cell adhesion molecules (CAM) play central roles in mediating immune and inflammatory responses. Leukocyte function-associated antigen (LFA-1, alpha(L)beta(2), and CD11a/CD18) and very late antigen (VLA-4, alpha(4)beta(1), and CD49d/CD29) are members of integrin-type CAM that are predominantly involved in leukocyte trafficking and extravasation. LFA-1 is exclusively expressed on leukocytes and interacts with its ligands ICAM-1, -2, and -3 to promote a variety of homotypic and heterotypic cell adhesion events required for normal and pathologic functions of the immune systems. VLA-4 is expressed mainly on lymphocyte, monocytes, and eosinophils, but is not found on neutrophils. VLA-4 interacts with its ligands VCAM-1 and fibronectin (FN) CS1 during chronic inflammatory diseases, such as rheumatoid arthritis, asthma, psoriasis, transplant-rejection, and allergy. Blockade of LFA-1 and VLA-4 interactions with their ligands is a potential target for immunosuppression. LFA-1 and VLA-4 antagonists (antibodies, peptides, and small molecules) are being developed for controlling inflammation and autoimmune diseases. The therapeutic intervention of mostly mAb-based has been extensively studied. However, due to the challenging relative efficacy/safety ratio of mAb-based therapy application, especially in terms of systemic administration and immunogenic potential, strategic alternatives in the forms of peptide, peptide mimetic inhibitors, and small molecule non-peptide antagonists are being sought. Linear and cyclic peptides derived from the sequences of LFA-1, ICAM-1, ICAM-2, VCAM-1, and FN C1 have been shown to have inhibitory effects in vitro and in vivo. Finally, understanding the mechanism of LFA-1 and VLA-4 binding to their ligands has become a fundamental basis in developing therapeutic agents for inflammation and autoimmune diseases. PMID- 11857638 TI - Binding interactions between peptides and proteins of the class II major histocompatibility complex. AB - The activation of helper T cells by peptides bound to proteins of the class II Major Histocompatibility Complex (MHC II) is pivotal to the initiation of an immune response. The primary functional requirement imposed on MHC II proteins is the ability to efficiently bind thousands of different peptides. Structurally, this is reflected in a unique architecture of binding interactions. The peptide is bound in an extended conformation within a groove on the membrane distal surface of the protein that is lined with several pockets that can accommodate peptide side-chains. Conserved MHC II protein residues also form hydrogen bonds along the length of the peptide main-chain. Here we review recent advances in the study of peptide-MHC II protein reactions that have led to an enhanced understanding of binding energetics. These results demonstrate that peptide-MHC II protein complexes achieve high affinity binding from the array of hydrogen bonds that are energetically segregated from the pocket interactions, which can then add to an intrinsic hydrogen bond-mediated affinity. Thus, MHC II proteins are unlike antibodies, which utilize cooperativity among binding interactions to achieve high affinity and specificity. The significance of these observations is discussed within the context of possible mechanisms for the HLA-DM protein that regulates peptide presentation in vivo and the design of non-peptide molecules that can bind MHC II proteins and act as vaccines or immune modulators. PMID- 11857639 TI - Therapeutic ultrasound: its application in drug delivery. AB - Ultrasound is best known for its imaging capability in diagnostic medicine. However, there have been considerable efforts recently to develop therapeutic uses for it. The purpose of this review is to summarize some of the recent advances made in the area of therapeutic ultrasound as they relate to drug delivery. In particular, this review will focus on the applications of ultrasound to enhance the delivery and effect of three distinctive therapeutic drug classes: chemotherapeutic, thrombolytic, and gene-based drugs. In addition, ultrasound contrast agents have been recently developed for diagnostic ultrasound. New experimental evidence suggests that these contrast agents can be used as exogenous cavitation nuclei for enhancement of drug and gene delivery. Thus, brief review of this new class of agents and their roles in drug delivery will also be provided. By comparison to diagnostic ultrasound, progress in therapeutic use of ultrasound has been somewhat limited. The recent successes in ultrasound related drug delivery research positions ultrasound as therapeutic tool for drug delivery in the future. PMID- 11857641 TI - Capillary electrophoresis for the analysis of glycosaminoglycans and glycosaminoglycan-derived oligosaccharides. AB - Glycosaminoglycans are a family of polydisperse, highly sulfated complex mixtures of linear polysaccharides that are involved in many life processes. Defining the structure of glycosaminoglycans is an important factor in elucidating their structure-activity relationship. Capillary electrophoresis has emerged as a highly promising technique consuming an extremely small amount of sample and capable of rapid, high-resolution separation, characterization and quantitation of analytes. Numerous capillary electrophoresis methods for analysis of intact glycosaminoglycans and glycosaminoglycan-derived oligosaccharides have been developed. These methods allow for both qualitative and quantitative analysis with a high level of sensitivity. This review is concerned with separation methods of capillary electrophoresis, detection methods and applications to several aspects of research into glycosaminoglycans and glycosaminoglycan-derived oligosaccharides. The importance of capillary electrophoresis in biological and pharmaceutical samples in glycobiology and carbohydrate biochemistry and its possible applications in disease diagnosis and monitoring chemical synthesis are described. PMID- 11857642 TI - Analysis of glycosaminoglycan-derived disaccharides in biologic samples by capillary electrophoresis and protocol for sequencing glycosaminoglycans. AB - Glycosaminoglycans are biologically significant carbohydrates which either as free chains (hyaluronan) or constituents of proteoglycans (chondroitin/dermatan sulfates, heparin, heparan sulfate and keratan sulfate) participate and regulate several cellular events and (patho)physiological processes. Capillary electrophoresis, due to its high resolving power and sensitivity, has been successfully used for the analysis of glycosaminoglycans. Determination of compositional characteristics, such as disaccharide sulfation pattern, is a useful prerequisite for elucidating the interactions of glycosaminoglycans with matrix effective molecules and, therefore, essential in understanding the biological functions of proteoglycans. The interest in the field of characterization of such biologically important carbohydrates is soaring and advances in this field will signal a new revolution in the area of glycomics equivalent to that of genomics and proteomics. This review focuses on the capillary electrophoresis methods used to determine the disaccharide pattern of glycosaminoglycans in various biologic samples as well as advances in the sequence analysis of glycosaminoglycans using both chromatographic and electrophoretic techniques. PMID- 11857643 TI - Analysis of glycoproteins and the oligosaccharides thereof by high-performance capillary electrophoresis-significance in regulatory studies on biopharmaceutical products. AB - This review describes the recent development in the analysis of glycoproteins using capillary electrophoresis with various separation techniques, and focuses especially on the analysis of recombinant glycoprotein pharmaceuticals. We include the analysis of glycoprotein multiforms (ie glycoform) as well as glycan analysis. The relationship between glycoprotein multiforms and oligosaccharide distributions in a glycoprotein sample is also discussed. Further, recent development in capillary electrophoresis-mass spectrometry is described. PMID- 11857644 TI - Structure analysis of lipoglycans and lipoglycan-derived carbohydrates by capillary electrophoresis and mass spectrometry. AB - Lipoglycans (lipopolysaccharides, lipoarabinomannans and glycolipids) are unique components of the cell membrane of all cells and the envelope of many bacteria. They play important roles in determining cell-environment interactions, which, however, are only partly understood due to incomplete description of their structural components, lipids and glycans. Capillary electrophoresis is an analytical technique of high separation efficiency and minimum sample requirements and has successfully been used for the analysis of several molecules of biological importance: proteins, nucleic acids and glycoconjugates. In the last years, a few applications of capillary electrophoresis to the analysis of lipoglycans have been reported. Analysis of lipoglycans involves the study of two parameters: intact molecules and carbohydrate parts. The conjunction of capillary electrophoresis and mass spectroscopy not only enhances the detection sensitivity, but also provides structural information on these structurally complex molecules. The interest in the field is rising and the results from the exact determination on the lipoglycan structure are expected to improve our understanding of the molecular mechanism of lipoglycan binding to proteins and cells of host organisms as well as their relationship to the virulence and pathogenesis of bacteria. In this report, an overview of the capillary electrophoresis methods used to analyze and characterize the intact lipoglycans as well as their carbohydrate parts is presented. PMID- 11857645 TI - Analysis of intact heparin by capillary electrophoresis using short end injection configuration. AB - A capillary electrophoresis method for the analysis of intact heparin was developed using a phosphate buffer and a fused silica capillary. Operational parameters such as pH and concentration of the running buffer were investigated. The short end injection configuration permitted a gain on peak efficiency, on the analysis time and on the repeatability of both migration times and peak areas, through a reduction of the migration distance. Moreover, the beneficial effect of the presence of sodium chloride in the heparin sample on the peak efficiency was demonstrated and the influence of the salts on the conformation of the heparin was discussed. The optimized method (short end injection configuration, 50mM phosphate buffer pH 3, heparin sample prepared in 10 g/L NaCl solution) was validated in terms of linearity, reproducibility and specificity according to ICH requirements. PMID- 11857646 TI - Employment of detergent-tag/solute interactions in capillary electrophoresis of neutral polysaccharides. AB - Neutral and inherently immobile polysaccharides are induced to migrate in an electric field through interactions with a detergent added to the electrophoretic electrolyte buffer. Before analysis the polysaccharides are converted to fluorescent derivatives to enable detection, but choice of a tag can also be utilized for modulation of the electrophoretic mobility. Three cases are discussed and exemplified, namely detergent-solute, detergent-solute+tag, and detergent-tag interactions. Anionic as well as cationic surfactants were exploited along with different derivatization reagents. Depending on the approach chosen, different kinds of information about sample composition and distribution(s) can be obtained, including degree of substitution, distribution of molecular weight (obtained in free solution without sieving media) and polymer conformation. A shift in polymer conformation upon a change in solvent composition can be monitored. PMID- 11857647 TI - Determination of molecular mass of acidic polysaccharides by capillary electrophoresis. AB - We developed a method for the determination of molecular mass of acidic polysaccharides based on their high-resolution separation by capillary electrophoresis. Polymers of N-acetylneuraminic acid (NeuAc) and polysulfated hyaluronic acid were separated into their molecular species up to 100 mono- and 20 disaccharide units, respectively. The relationship between the molecular mass of NeuAc-polymer and their electrophoretic mobilities showed good linearity, and was applied to the determination of molecular masses of larger NeuAc species unresolved by capillary electrophoresis under the same conditions. In the first step, the standard curve for the determination of molecular mass was constructed from the relationship between electrophoretic mobility and molecular mass. Subsequently, the mobility was extrapolated to the standard curve, and the molecular mass was calculated. Five different preparations of NeuAc polymers having different molecular masses showed smaller values than those determined by conventional chromatographic techniques. Further, molecular mass determined by the present method correlated with number-average molecular mass. The methodology presented here was applied to the determination of molecular mass of polysulfated hyaluronic acid. The data indicated that native hyaluronic acid was extensively degraded during sulfonation reaction. PMID- 11857648 TI - A capillary zone electrophoresis method for determining N-acetylneuraminic acid in glycoproteins and blood sera. AB - A simple capillary zone electrophoresis (CZE) method for the determination of the content of the major sialic acid form N-acetylneuraminic acid (Neu5Ac) in glycoproteins was established. The present method utilizes a simplified hydrolysis-purification procedure consisting of mild acid hydrolysis (25 mM trifluoroacetic acid for 2h at 80 degrees C) to release Neu5Ac and ultrafiltration on Centricon-3 membrane to remove the obtained asialoglycoproteins and other macromolecules present in biologic samples. Derivatization with benzoic anhydride at 80 degrees C for 20 min resulted in complete conversion of Neu5Ac to per-O-benzoylated Neu5Ac. CZE analysis was performed using the operating buffer 25mM phosphate, pH 3.5, containing 50% (v/v) acetonitrile as organic modifier at 30 kV, and detection of the per-O-benzoylated Neu5Ac at 231 nm. The method showed excellent repeatability (RDS<1.98%) and a linearity range from 5 microg/mL to 5mg/mL with a detection limit of 2 microM. Application of the method to microanalysis of human alpha(1)-acid glycoprotein and blood serum samples showed excellent agreement with previously published values, suggesting a high precision for the developed CZE method. PMID- 11857649 TI - Nano-HPLC-mass spectrometry and MEKC for the analysis of oligosaccharides from human milk. AB - The separation of oligosaccharides derivatized with various esters of aminobenzoic acid by means of reversed-phase nano-HPLC (nHPLC) with on-line ESI mass spectrometry and off-line MALDI-TOF mass spectrometry as well as MEKC is described. For this purpose methyl, ethyl and butyl aminobenzoates and heptyloxyaniline were used as derivatization agents for homologous maltodextrins and oligosaccharides from human milk. Four different C(18) stationary phases were tested for this purpose because the type of stationary phase was shown to have a dramatic effect on the performance of the separation. Optimal results were obtained using n-butyl aminobenzoate as label and an encapsulated ODS stationary phase. The on-line coupling of nHPLC to ESI MS allowed to separate and identify various oligosaccharides from human milk. This technique enabled the exact attribution of the molecular structure to a signal in the chromatogram. In a second approach oligosaccharides were separated by nHPLC and subsequently fractionated. The fractions were analyzed by MALDI-TOF mass spectrometry. The results obtained by this approach confirmed the ESI MS data. An analogous separation profile was obtained by using sodium dodecyl sulfate in MEKC, which proves that the retention mechanisms of both techniques are identical. PMID- 11857650 TI - High-performance capillary electrophoretic analysis of hyaluronan and galactosaminoglycan-disaccharides in gastrointestinal carcinomas. Differential disaccharide composition as a possible tool-indicator for malignancies. AB - The glycosaminoglycans (GAGs) have documented implications for the growth and progression of malignant tumors. Gastrointestinal carcinomas (gastric, colon, rectum and pancreatic) are the most frequent malignancies occurring in human. GAGs, isolated from the tissues after digestion with papain, were analyzed by high-performance capillary electrophoresis (HPCE) following treatment with chondroitinase ABC. The composition of GAGs in disaccharides derived from the various gastrointestinal carcinomas was compared with those of normal tissues. We report that human gastrointestinal carcinomas are characterized by increased concentrations of GAGs, which have quite different disaccharide composition which, in turn, is associated with marked increase of non-sulfated (Delta(di) nonS) and 6-sulfated (Delta(di)-mono6S) Delta-disaccharides. Particularly, a 12 51-fold increase in Delta(di)-nonS and a 3-42-fold increase in Delta(di)-mono6S content characterize these carcinomas, while the 4-sulfated units (Delta(di) mono4S) showed a lower increase, about 0.5-1.5-fold. Moreover, the quantitation of hyaluronan (HA)-derived Delta-disaccharides (Delta(di)-nonS(HA)) also revealed a marked increase (1-12-fold) in the malignant tissues. On the other hand, the content of the chondroitinase ABC-resistant GAGs showed a low decrease, about 0.2 0.7-fold. The high amounts of hyaluronan (HA) produced by these carcinomas and the ectopic production of chondroitin sulphate (CS) proteoglycans, in which (Delta(di)-nonS) and (Delta(di)-mono6S) predominated, suggest a close relation between the content of these GAGs and the malignant phenotype, the metastatic ability and the survival time. PMID- 11857651 TI - Familiarity and dominance relations among female sooty mangabeys in the Tai National Park. AB - Dominance relationships of female sooty mangabeys have thus far been studied exclusively in captive groups. In captivity, adult females form a stable linear hierarchy as would be expected in species exhibiting strong contest competition. However, the same individuals do not exhibit other aspects of behavior that would be expected where contest competition occurs. For example, they show no kin-based alliances leading to hierarchies in which the members of each matriline occupy adjacent ranks. The goal of this study was to provide the first data on dominance relationships of sooty mangabey females in their natural environment in the Tai National Park, Ivory Coast. In our study group, adult females formed a linear dominance hierarchy. Aggression over food increased in food patches, as would be expected for species that experience contest competition. Moreover, females formed highly differentiated social relationships, showing particular affinities with females of adjacent rank. PMID- 11857652 TI - Mantled howler population of Hacienda La Pacifica, Costa Rica, between 1991 and 1998: effects of deforestation. AB - A survey of the mantled howler (Alouatta palliata) population on Hacienda La Pacifica, Guanacaste, Costa Rica, was done in July and August of 1998 to determine population parameters following deforestation due to major canal construction between 1990 and 1994. The survey was carried out in a manner identical to our 1991 survey and consisted of a single pass and two re-surveys of all forested areas of the farm. As canal construction effectively increased fragmentation of the habitat, we predicted decreased population and group size over this time. Results indicated that between the 1991 and 1998 survey, group size decreased but not significantly, and there were significantly fewer adult males and adult females per group. Population size, however, remained unchanged as there was an increase in animals in the immature age classes. An increase in the infant to adult female ratio suggests a stable or even expanding population, which could represent recovery from the initial disturbance of deforestation. Thus, despite changes in the forest and land use patterns, the area now appears to support the same number of howlers as found in previous surveys. PMID- 11857653 TI - Survival, growth, health, and reproduction following nursery rearing compared with mother rearing in pigtailed monkeys (Macaca nemestrina). AB - Nursery-reared primates do not experience psychological "maternal bonding" or immunological benefits of breast milk, so they are expected to be inferior to mother-raised monkeys in growth, health, survival, reproduction, and maternal abilities. Studies of nursery-reared monkeys support aspects of this prediction for infants deprived of social contact or raised in pairs. We present colony record data on 1,187 mother and 506 nursery-raised monkeys, 2-10 yr of age, living in mixed groups. We found no group differences in survival, growth, clinical treatments for disease or bite wounds, or pregnancy outcome and neonatal deaths. Nursery males given breeding opportunities produced an average of 24 offspring. In addition to 24-hr personnel present on every day of the year, we believe that three of our procedures account for differences between our results and other reports. Our infants received 1) intensive human handling, 2) daily social interaction in a playroom, and 3) success and failure experience during learning and cognitive testing. We do not advocate rearing primates without mothers, but we conclude that these procedures are sufficient for producing physical health and adaptive juvenile and adult social skills in nursery-raised monkeys. PMID- 11857656 TI - News and comment. PMID- 11857654 TI - Single-copy nuclear DNA sequences obtained from noninvasively collected primate feces. AB - Noninvasively collected primate feces have been shown to provide a useful source of mitochondrial DNA for sequencing and nuclear microsatellite DNA for size analysis. In this study, single-copy nuclear DNA sequences were obtained from noninvasively collected fecal samples of two species of wild tamarins, Saguinus fuscicollis and S. mystax, in the context of a project on the functional utility of color vision. Noninvasive genotyping of the X-linked opsin gene is important for future studies of selection and adaptation at this locus in a number of primate species. The wide range of techniques that can now be applied successfully to DNA extracted from feces introduces a broad spectrum of potential genetic studies that can be undertaken on primates, without the need for intrusive or invasive methods. PMID- 11857658 TI - Maurice Victor: 1920--2001. PMID- 11857659 TI - News and comment. PMID- 11857660 TI - Choosing words. PMID- 11857661 TI - Subjective and objective incontinence 5 to 10 years after Burch colposuspension. AB - The outcome of incontinence surgery was studied using a questionnaire, a 24-hour pad test (24hPT), and a stress test (ST). Five to 10 years after a Burch colposuspension, 111 patients were asked to complete the Bristol Female Urinary Tract Symptom Questionnaire (BFLUTS) and to perform a 24hPT and a ST. Eighty-two patients completed the questionnaire and 71 and 69 patients performed the stress and pad tests, respectively. Seventy-three percent of the patients did not leak during the ST and 75% of the patients were not leaking during the 24hPT. Seventy three percent of the patients stated that they were at least occasionally stress or urge incontinent and 62% stated that they were both stress and urge incontinent. However, only 24% of the stress incontinent and 28% of the urge incontinent patients found their incontinence to be "quite a problem" or "a serious problem." Patients leaking urine only "occasionally," "once a week," leaking "drops," and finding the leakage to be "a bit of a problem" had median leakage 0g during ST and 24hPT. Patients who reported the leakage to occur "sometimes" "most or all of the time" and who found the leakage to be "a bit, quite, or a serious problem" accounted for 20 to 30% of all patients, as did patients leaking during objective tests. Objective tests revealed leakage to occur less frequently compared with self-reported leakage. The BFLUTS questionnaire revealed leakage to occur with varying frequency, amount, and bother. Leakage occurring seldom, of small amount and bother may be of minor clinical importance. PMID- 11857662 TI - Data from frequency-volume charts versus filling cystometric estimated capacities and prevalence of instability in men with lower urinary tract symptoms suggestive of benign prostatic hyperplasia. AB - The aim was to examine associations of filling cystometric estimated compliance, capacities, and prevalence of bladder instability with data from frequency-volume charts in a well-defined group of men with lower urinary tract symptoms (LUTS) suggestive of benign prostatic hyperplasia (BPH). Men with LUTS suggestive of BPH were included if they met the criteria of the International Consensus Committee on BPH, i.e., they voided more than 150 mL during uroflowmetry, their residual volume and prostate size were estimated, and they completed frequency-volume charts correctly. From the frequency-volume charts, voiding habits, and fluid intake in the daytime and at night were evaluated. Filling cystometric studies were performed in these men as well. Decreased compliance was an exceptional finding. Cystometric capacity and especially effective capacity (cystometric capacity minus residual volume) corresponded significantly with the maximum voided volume on the frequency-volume charts. Effective capacity was almost twice as high as the average voided volume. Minimum voided volume on frequency-volume charts was not related to filling cystometric data. The presence of instability in the supine or sitting position or in both positions was not significantly associated with smaller voided volumes, higher nocturia, or diuria. Filling cystometric capacities were strongly associated with maximal and mean voided volumes derived from frequency-volume charts. The presence of detrusor instability during filling cystometry did not significantly affect voided volumes, diuria, or nocturia PMID- 11857663 TI - Comparison of long-term results of transurethral incision of the prostate with transurethral resection of the prostate, in patients with benign prostatic hypertrophy. AB - One hundred patients with benign prostatic hypertrophy (BPH) were randomized to transurethral incision (TUIP) or transurethral resection of the prostate (TURP). The average prostate weight before operation was not more than 30.0 g. Indications for the operations were based on the disease history, physical examination, digital rectal examination, laboratory values, and pressure-flow examination. All operations were performed with patients under spinal anesthesia. TUIP was performed with a Collins knife, and TURP was performed with a resectoscope. Follow-up was performed 24 months after the operations. After treatment there were statistically significant daytime and nocturnal reduction in voiding frequencies of 2.9 and 1.7, respectively, after TUIP, and 2.0 and 1.5 after TURP. In both groups, there occurred significantly better maximal flow rate from 7.6 mL/s to 16.9 mL/s in group I and from 6.9 mL/s to 17.6 mL/s in group II. The mean values of linearized passive urethral resistance relation in both groups significantly decreased from 3.6 +/- 0.6 to 1.0 +/- 0.5 after TUIP and from 3.9 +/- 04 to 1.4 +/- 0.5 after TURP. The TUIP procedure is effective and safe for patients with a small number of complications. PMID- 11857664 TI - Development of a non-invasive strategy to classify bladder outlet obstruction in male patients with LUTS. AB - To diagnose bladder outlet obstruction in male patients with lower urinary tract symptoms (LUTS), it is necessary to measure the bladder pressure via a transurethral (or suprapubic) catheter. This procedure incurs some risk of urinary tract infection and urethral trauma and is sometimes painful to the patient. We developed an external condom catheter to measure non-invasively the bladder pressure and developed a strategy to classify bladder outlet obstruction (BOO) based on this measurement. Seventy-five patients with a wide range of urological diagnoses underwent a pressure-flow study followed by a non-invasive study. We tested five different strategies to classify the patients using the provisional International Continence Society (ICS) method for definition of obstruction as the gold standard. Leakage of the external catheter occurred in eight (40%) of the first 20 tested patients. In the remaining 55 patients, only five (9%) of the measurements failed because of leakage. Of the 75 patients, 56 were successfully tested non-invasively. According to the ICS nomogram, the PFS showed that 22 of these patients were non-obstructed, 12 patients were equivocal, and 22 patients were obstructed. Ten of these 56 patients strained, and we found that the relatively high abdominal pressures in these patients were not reflected in the externally measured bladder pressure. Of the remaining 46 patients, 12 of 13 non-obstructed patients and 30 of 33 combined equivocal and obstructed patients could be correctly classified. We developed a simple, non-invasive classification strategy to identify BOO in those male patients who did not strain during voiding. PMID- 11857665 TI - Urge incontinence and impaired detrusor contractility in the elderly. AB - Among the elderly, both urge incontinence and elevated residual urine are common. When they occur together, they present a challenging clinical problem, called detrusor hyperactivity with impaired contractile function (DHIC). Impaired detrusor contractility has two aspects: elevated post-void residual urine volume and reduced detrusor contraction strength. Geriatric urge incontinence, especially in combination with reduced bladder sensation, is associated with specific cortical abnormalities: frontal and global cortical underperfusion and cognitive impairment. We have investigated, in 73 elderly incontinent patients, whether either aspect of impaired contractility is associated with urge incontinence, reduced sensation or these cortical abnormalities. For post-void residual urine, there are no significant associations. Detrusor contraction strength, however, is significantly increased (not impaired) if there is urge incontinence, reduced bladder sensation, or cortical underperfusion. Thus, DHIC appears to be a coincidental occurrence of two common conditions with different etiological factors. PMID- 11857666 TI - Does post-voiding residual volume get less as mobility improves in a rehabilitation ward for older adults? AB - Impaired bladder emptying is common in frail older adults. This study tests the hypothesis that more complete bladder emptying is associated with better mobility in a rehabilitation ward for older adults. Consecutive admissions to a rehabilitation ward for older adults were considered for inclusion in the study in the week after admission to the ward. Exclusion criteria were cognitive impairment such that consent could not be obtained, non-English speaking, or presence of an indwelling urinary catheter. A post-voiding residual (PVR) bladder volume and Rivermead Mobility Index (RMI) were completed for subjects who gave consent, on a weekly basis until discharge. The BladderScan BVI 3000, Diagnostic Ultrasound, instrument was used. Statistical analysis was by a general linear mixed model. In the study period, 114 people were admitted and 57 approached for consent. Twenty-four people gave consent. Fifty percent of participants had at least one PVR greater than 100 mL. One person was found to be in urinary retention, with a PVR of greater than 700 mL, and was excluded from further analysis. The PVR increased by 1.2 mL (95% CI-4.6 to 7) for each unit improvement in the RMI. This study suggests that PVR does not decline with improvement in mobility in older adults receiving inpatient rehabilitation. PMID- 11857667 TI - Similarities and differences in female and male rat voiding. AB - We measured in adult rats, under anaesthesia, bladder pressure by transvesical cystometry and flow rate by an ultrasound transducer in the distal urethra. The urinary flow was discontinuous in both sexes. No difference between the sexes in bladder pressure oscillations or in non-oscillatory voiding was found but during the oscillatory activity there was a difference in the relationship between bladder pressure and urinary flow. In the female, the bladder pressure decreased when the flow started and increased when the flow decreased resembling species whose urinary flow is continuous. Basically the flow was stable but it was divided into periods of variable duration by full or partial closure of urethral sphincter. In the male rat, the oscillatory flow consisted of short, fast spikes occurring just before the bladder pressure reached the maximum, after which the flow spike decreased slowly. Overall, no differences were seen in bladder pressure data between the genders. However, the maximal flow rate was lower and micturition time was shorter in female rats. When we recorded occasionally occurring micturitions without high-frequency oscillations of intraluminal pressure (IPHFOs) (non-oscillatory voiding), no differences between the genders were seen. The difference during oscillatory voiding between male and female rat can be understood against anatomical and hormonal backgrounds, and by the relative role of rhabdosphincter, which did not activate during non-oscillatory voidings when no differences were detected. PMID- 11857668 TI - Cholinergic and purinergic contribution to the micturition reflex in conscious rats with long-term bladder outlet obstruction. AB - The urethra of female Wistar rats was partially obstructed for 15 weeks. The effects of atropine (1 mg/kg i.v.), suramin (100 mg/kg i.v.), and a combination of atropine and suramin on the peak micturition pressure (MP) were compared during cystometry in conscious rats controls or subjected to outlet obstruction. On the isolated bladder dome, we studied the inhibitory effect of 1 micromol/L atropine, 1 mmol/L suramin, and the combination of the two drugs on contractions induced by electrical field stimulation (EFS). We studied also the contractile response to 80 mmol/L KCl and the concentration-response curves to noradrenaline, phenylephrine, and carbachol on the bladder dome and bladder neck and alpha, beta methylene adenosine triphosphate on the bladder dome. In conscious rats, the MP, bladder capacity, and micturition volume were significantly higher in obstructed rats than in controls. Suramin induced the same inhibition in the two groups of animals (-30.7 +/- 13.3% in controls and -29.2 +/- 8.5% in obstructed rats). Atropine decreased the MP, but this effect was twofold greater in obstructed animals (-28.1 +/- 3.1% and -65.1 +/- 6.9% in control and obstructed animals, respectively). However, the combined effect of atropine and suramin was additive in controls but not in obstructed (-56.7 +/- 5.4% and -55.9 +/- 9.4%, respectively). Similar results were obtained in vitro using 1 micromol/L atropine and 1 mmol/L suramin. In the obstructed bladder dome and bladder neck, we found a great reduction in KCl- and carbachol-induced contractility but no difference in the response to EFS. Responses to noradrenaline and phenylephrine were moderately reduced in the bladder neck only, whereas responses to alpha, beta-methylene adenosine triphosphate in the bladder dome were not reduced except at the concentration of 300 micromol/L. We conclude that long-term obstruction in rats could induce cholinergic nerve fiber proliferation as suggested by the decrease in M(3) muscarinic receptor contractility (desensitization) and by a greater sensitivity of the MP to atropine. PMID- 11857669 TI - Role of sorbitol in the up-regulation of urinary bladder M(2) muscarinic receptors in streptozotocin-induced diabetic rats. AB - This study investigated the role of sorbitol, a metabolic product of glucose, in the pathogenesis of rat diabetic cystopathy. Three-month-old male Wistar rats were divided into four groups: 1) normal controls; 2) rats rendered diabetic by streptozotocin; 3) rats fed with glucose; and 4) rats injected with sorbitol. The M(2) muscarinic receptor (M(2)-mAChR) protein and mRNA densities of the bladder tissue were measured by Western immunoblot and Northern blot, respectively. The streptozotocin-induced diabetic rats were then treated with ONO-2235, an aldose reductase inhibitor. The bladder M(2)-mAChR protein and mRNA were compared between the treated and untreated diabetic rats. The densities of M(2)-mAChR protein and mRNA in the bladder tissue were significantly increased in diabetic rats, and rats given either glucose or sorbitol (increases in receptor protein: 27.3 +/- 3.3, 19.8 +/- 2.3, and 18.0 +/- 2.1%; increases in mRNA: 39.6 +/- 3.7, 33.1 +/- 2.9, and 20.2 +/- 2.2%, respectively). When diabetic rats were treated with ONO-2235, the increases in bladder M(2)-mAChR protein and mRNA were significantly alleviated. The findings suggest that sorbitol plays a role in the pathogenesis of diabetic cystopathy in rats rendered diabetic by streptozotocin. Aldose reductase inhibitors may be useful in the treatment and prevention of diabetic cystopathy. PMID- 11857670 TI - Effect of doxazosin on rat urinary bladder function after partial outlet obstruction. AB - Hypoxia induced by partial outlet obstruction is believed to play a major role in both the hypertrophic and degenerative effects of partial outlet obstruction. Doxazosin (dox) is a clinically effective alpha-adrenergic antagonist used in the treatment of symptomatic benign prostatic hyperplasia (BPH). Although the major therapeutic effect of the agent is believed to occur on the smooth muscle components of the prostate by reducing prostatic urethral resistance and thus improving emptying, dox may have part of its clinical action via effects mediated by other actions, including via spinal alpha-adrenergic receptors or direct effects on the bladder, possibly via inhibition of vascular alpha receptors. The specific aim of the current study was to determine whether dox pretreatment on rats affects blood flow to the bladder and reduces the level of bladder dysfunction induced by partial outlet obstruction. In part 1, eight rats were separated into two groups of four rats each. Group 1 received oral administration of dox (30 mg/kg) for 4 weeks; group 2 received vehicle (5% dimethyl sulfoxide). After 4 weeks of treatment, blood flow studies were performed using fluorescent microspheres and the bladders excised, frozen, and submitted to Interactive Medical Technologies (IMT) for blood flow analysis. In part 2, 32 adult male rats were separated into four groups of eight rats each. Groups 1 and 2 received oral administration of dox (30 mg/kg) for 4 weeks, groups 3 and 4 received vehicle (5% dimethyl sulfoxide). At 4 weeks, the rats in groups 1 and 3 received partial outlet obstructions and treatment continued for an additional 2 weeks. After 6 weeks of treatment (total), each rat was anesthetized, the bladder excised, weighed, and isolated strips mounted and contractility studies performed. 1) Four weeks pretreatment of rats with dox increased blood flow to the bladder in both the control and obstructed groups. 2) Partial outlet obstruction induced a mild decrease in blood flow. 3) The magnitude of the increased bladder weight in the vehicle-treated obstructed group was significantly greater than in the dox treated obstructed group. 4) Partial outlet obstruction resulted in significant decreases in the contractile response to field stimulation in both treated and non-treated rats. The magnitude of the decreased response was significantly greater in the non-treated rats. 5) The response to potassium chloride was significantly reduced by partial outlet obstruction in the vehicle-treated group but not in the dox-treated group. 6) The time to maximal tension was significantly increased in response to carbachol, adenosine triphosphate, and potassium chloride. However, the magnitude of the increase was significantly greater for the vehicle-treated obstructed groups stimulated by potassium chloride than for the dox-treated groups. Dox treatment of rats increased blood flow to the bladder and reduced the severity of the response to partial outlet obstruction. These beneficial effects would be due to pharmacological effects on alpha-adrenergic systems outside those present in the prostate. These include effects on blood flow to the bladder, effects on the micturition centers of the central nervous system, spinal reflexes, and alpha-adrenergic receptors in the urethra and bladder. PMID- 11857671 TI - The standardisation of terminology of lower urinary tract function: report from the Standardisation Sub-committee of the International Continence Society. PMID- 11857672 TI - The standardisation of terminology in nocturia: report from the Standardisation Sub-committee of the International Continence Society. PMID- 11857673 TI - Growth and development of Encarsia formosa (Hymenoptera: Aphelinidae) in the greenhouse whitefly, Trialeurodes vaporariorum (Homoptera: Aleyrodidae): effect of host age. AB - The tiny parasitoid wasp, Encarsia formosa, has been used successfully to control greenhouse whiteflies (GHWFs) in greenhouses in many countries throughout the world. Therefore, there has been considerable interest in developing methods for artificially rearing this wasp. However, little information is available concerning the regulation of its development including the host-parasitoid interactions that are required for the parasitoid to complete its life cycle. Here we confirm that parasitoid developmental rates differ significantly based upon the host instar parasitized. Development was faster when 3rd and 4th instar GHWFs were offered for parasitization than when 1st or 2nd instars were used. Our results show that it is primarily the embryo and the first two parasitoid instars that exhibit prolonged developmental times when 1st and 2nd instar whiteflies are parasitized. Although percent emergence was not affected by host age at the time of parasitization, adult longevity as well as adult emergence pattern varied greatly depending upon the instar parasitized. When 3rd and 4th instar GHWFs were selected for oviposition, adult wasps lived significantly longer than when 1st or 2nd instars were used; also, there was a sharp emergence peak on the 2nd day after emergence was first observed (reduced or absent when 1st or 2nd instar GHWFs were parasitized) and the emergence period was reduced from between 8 and 11 days to 5 days. In general, the younger the host instar parasitized, the less synchronous was parasitoid development. Previous reports that E. formosa will not molt to the 2nd instar until the host has reached its 4th instar were not confirmed. When 1st instar host nymphs were parasitized, 2nd instar parasitoids were detected in 3rd instar hosts. Importantly, however, no matter which instar was parasitized, the parasitoid never molted to its last instar until the host had reached Stage 5 of its last instar, a stage in which host pharate adult formation has been initiated. It appears, then, that a condition(s) associated with host pharate adult formation is required for the parasitoid's final larval molt. Results reported here should facilitate the development of in vitro rearing systems for E. formosa. PMID- 11857674 TI - Stimulation of sex pheromone production by PBAN-like substance in the pine caterpillar moth, Dendrolimus punctatus (Lepidoptera: Lasiocampidae). AB - Sex pheromone production in the female pine caterpillar moth, Dendrolimus punctatus is controlled by a PBAN-like substance located in the head of female moth. Pheromone titer was significantly decreased by decapitation of female moth, and restored by injection of either Hez-PBAN or head extract prepared from male or female moth. Stimulation of pheromone production by head extract followed a dose-dependent pattern from 0.5 to at least 4 head equivalent. A gland in vitro assay was used to study the relationship between gland incubation time and pheromone production as well as calcium involvement in the stimulation of pheromone production by head extract. Maximum pheromone production was occurred at 60 min after pheromone gland was incubated with two equivalents of head extracts. In vitro experiments showed that the presence of calcium in the incubation medium was necessary for stimulation of pheromone production. The calcium ionophore, A 23187, alone stimulated pheromone production. The pheromone components (Z,E)-5,7-dodecadienol and its acetate and propionate were produced in these experiments but in addition to the aldehyde, (Z,E)-5,7-dodecadienal was also found. This indicates that females are capable of producing four oxygenated functional groups. The PBAN-like substance control of the pheromone biosynthetic pathway was investigated by monitoring the incorporation of the labeled precursor into both pheromone and pheromone intermediates. PMID- 11857675 TI - Characterization of cockroach (Periplaneta americana) fat body phospholipase A(2) activity. AB - A phospholipase has been identified in the fat body of the American cockroach, Periplaneta americana, which removes fatty acid from the sn-2 acyl position of an artificial substrate. The enzyme has been characterized using a crude preparation obtained by low-speed centrifugation of the homogenized tissue. With 1 hexadecanoyl-2-(1-pyrenedecanoyl)-sn-glycero-3-phosphocholine as the substrate, the K(m) has been estimated to be 1.17 microM and the v(max) 113.5 pmol/min/mg protein. The phospholipase has a pH optimum close to 7 and shows maximal activity at 50 degrees C. Activity of the phospholipase has been determined in cytosolic and plasma membrane fractions. The specific activity of the latter fraction is approximately twice that of the cytosol. The enzyme in both fractions is Ca(2+) independent. Arch. PMID- 11857676 TI - Glutathione S-transferase isoenzymes in the two-spot ladybird, Adalia bipunctata (Coleoptera: Coccinellidae). AB - Isoenzymes of glutathione S-transferase (GST) in adult Adalia bipunctata, an aphidophagous predator, were studied. Cytosolic GST activity was studied in each beetle developmental stage. The highest activities towards both 1-chloro-2,4 dinitrobenzene (CDNB) and 2,4-dinitro-1-iodobenzene (DNIB) occurred in adults. The enzyme distribution was investigated in adults. While most of the enzymatic activity was found in the abdomen (40-50 and 34-63% respectively) using several concentrations of both CDNB and DNIB, significant differences were observed for the head and the thorax depending on the substrate. Activities were more abundant in the thorax with DNIB (37-47%) compared to the 13-19% obtained with CDNB. Some GST activity was also detected in the elytra. GSTs were purified by epoxy activated Sepharose 6B affinity chromatography and applied to an HPLC column to determine the native molecular weight (69 kDa). Three isoenzymes were separated by chromatofocusing at pH ranges 7-4. Three bands with molecular mass from 23 to 26 kDa were visualised on SDS-PAGE. Their isoelectric points were 6.66, 6.36, and 6.21. The substrate specificities and the kinetic parameters (Vm and Km) of the isoenzymes showed large differences depending on the isoenzyme. Arch. PMID- 11857677 TI - Absorption and tissue distribution of cholesterol in Manduca sexta. AB - In Manduca sexta larvae, radioactive free cholesterol is absorbed directly from the midgut into mucosal cells where it is stored both in the free form (87% in males and 93% in females) and esterified form (13% in males and 7% in females). Subsequently, cholesterol is transported to fat body via lipophorin in the hemolymph exclusively in the free form. In fat body, the distribution of cholesterol between the free and esterified form varied significantly between genders and developmental stages. Except for the larval stage, males and females were able to store cholesterol in both free and esterified forms in the fat body and in the adult stage cholesterol ester accounted for more than 75% of the stored cholesterol. Placement of radioactive cholesterol at different locations in the gut-foregut, midgut, and hindgut-demonstrated that the midgut is the site where cholesterol is absorbed and released into the hemolymph. Cholesterol labeled lipophorin injected into larval hemolymph was cleared from the hemolymph with a half-life of 10.2 h. After 17 h, most of the cleared radioactivity was recovered in the fat body (38%). Arch. PMID- 11857678 TI - Reaction of mouse brain oligodendrocytes and their precursors, astrocytes and microglia, to proinflammatory mediators circulating in the cerebrospinal fluid. AB - The response of glial cells to the acute intracerebroventricular administration of interferon-gamma, and of this cytokine combined with the endotoxin lipopolysaccharide or with tumor necrosis factor-alpha, was investigated in the brain of adult mice over a time course of 1 week. Oligodendrocytes were identified by immunocytochemistry, using O4 to label their precursors and 2',3' cyclic nucleotide 3'-phosphohydrolase as marker of mature cells. Astrocytes were labeled by glial fibrillary acidic protein immunoreactivity and microglial cells by tomato lectin histochemistry. Compared with ovalbumin-injected control cases, all cytokine treatments caused a marked decrease of immunostained mature oligodendrocytes in the brain since 1 day postinjection. O4+ oligodendrocyte precursors increased instead progressively from 2 to 7 days. Astrocytes, markedly activated by cytokine treatments, also exhibited a progressive quantitative increase from 2 days onward. Activation and proliferation of microglial cells were instead most evident at 24 h postinjection. Such glial responses to interferon-gamma injections were especially marked in the periventricular brain parenchyma and were enhanced by coadministration of lipopolysaccharide or tumor necrosis factor-alpha. The findings show that a pulse of proinflammatory mediators in the cerebrospinal fluid affects mature oligodendrocytes, concomitantly with the early appearance of activated microglia, and that such reactions are rapidly followed by an increase of oligodendrocyte precursors paralleled by astrocytic activation. The data, which allowed dissecting the events elicited in glial cell populations by inflammatory mediators via the cerebrospinal fluid, indicate that these molecules elicit in vivo a toxic effect on mature oligodendrocytes and a stimulation of their precursors in the adult brain. PMID- 11857679 TI - Transforming growth factor-beta and forskolin attenuate the adverse effects of long-term Schwann cell denervation on peripheral nerve regeneration in vivo. AB - Transforming growth factor-beta (TGF-beta) plays a central role in the regulation of Schwann cell (SC) proliferation and differentiation and is essential for the neurotrophic effects of several neurotrophic factors (reviewed by Unsicker and Krieglstein, 2000; Unsicker and Strelau, 2000). However, its role in peripheral nerve regeneration in vivo is not yet understood. Our studies were carried out to characterize (1) the effects of duration of regeneration, and chronic SC denervation on the number of tibial (TIB) motor neurons that regenerated axons over a fixed distance (25 mm into distal common peroneal [CP] nerve stumps), and (2) the effect of in vitro incubation of 6-month chronically denervated sciatic nerve explants with TGF-beta and forskolin on their capacity to support axonal regeneration in vivo. TIB--CP cross-suture in Silastic tubing was used, and regeneration into 0-24-week chronically denervated CP stumps was allowed for either 1.5 or 3 months. Chronically denervated rat sciatic nerve explants (3 x 3 mm(2)) were incubated in vitro with either DMEM and 15% fetal calf serum (D-15) plus TGF-beta/forskolin or D-15 alone for 48 h and placed into a 10-mm Silastic tube that bridged the proximal and distal nerve stumps of a freshly cut TIB nerve. The number of tibial motor neurons that regenerated axons through the explants and 25 mm into the distal nerve stump after 6 months, and TIB regeneration into the CP nerve stumps, were assessed using retrograde tracers, fluorogold, or fluororuby. We found that all tibial motor neurons regenerate their axons 25 mm into 0-4-week denervated CP nerve stumps after a regeneration period of 3 months. Reducing regeneration time to 1.5 months and chronic denervation, reduced the number of motor neurons that regenerated axons over 25 mm. Exposure of 6-month denervated nerve explants to TGF-beta/forskolin increased the number of motor neurons that regenerated through them from 258 +/-13; mean +/ SE to 442 +/- 22. Hence, acute treatment of atrophic SC with TGF-beta can reactivate the growth-permissive SC phenotype to support axonal regeneration. PMID- 11857680 TI - Identification of Golli and myelin basic proteins in human brain during early development. AB - The myelin basic protein gene (Mbp) encodes for the major myelin structural proteins and it is included in the Golli-Mbp gene complex. Previously, we observed MBP-like proteins in the human central nervous system (CNS) at developmental stages preceding myelination. In an effort to distinguish between Golli (HOG5 and HOG7) and MBP mRNAs and to determine their spatiotemporal distribution, we performed in situ hybridization using two human Golli specific probes: one corresponding to exon 5a absent from all MBP transcripts, and the other corresponding to exon 5c specific for HOG5. HOG7 transcript was observed first, in 5 gestational week-old embryos, whereas both Golli transcripts were detected at 6-7 weeks gestation in the proliferative zones of the entire CNS. Golli proteins immunoreactivity was observed in microglia and early neurons of the developing telencephalon. During midgestation (17-22 weeks gestation), at the onset of myelination, MBP and Golli mRNAs were observed in the telencephalic subventricular zone and occasionally in the future cerebral cortex. Developmental expression of the human Golli-Mbp indicates that the two Golli proteins have different onset of expression, distribution and possibly function. These results support the hypothesis that at least one of them, HOG7, may be involved in the regulation of early neurogenesis, while both may have additional, still undefined function at the onset of myelination. PMID- 11857681 TI - Juxtavascular microglia migrate along brain microvessels following activation during early postnatal development. AB - Some parenchymal microglia in mammalian brain tissues, termed "juxtavascular microglia," directly contact the basal lamina of blood vessels; however, the functional consequences of this unique structural relationship are unknown. Here we used a rat brain slice model of traumatic brain injury to investigate the dynamic behavior of juxtavascular microglia following activation. Juxtavascular microglia were identified by confocal 3D reconstruction in tissue slices stained with a fluorescent lectin (FITC-IB4) that labels both microglia and blood vessel endothelial cells. Immunolabeling confirmed that juxtavascular cells were true parenchymal microglia (OX42+, ED2-) and not perivascular cells or pericytes. Time lapse imaging in live tissue slices revealed that activating juxtavascular microglia withdraw most extant branches but often maintain contact with blood vessels, usually moving to the surface of a vessel within 1-4 h. Subsequently, some microglia migrate along the parenchymal surface of vessels, moving at rates up to 40 microm/h. Activated juxtavascular microglia sometimes repeatedly extend veil-like protrusions into the surrounding tissue, consistent with a role in tissue surveillance. Juxtavascular cells were twice as likely as nonjuxtavascular cells to be locomotory by 10 h in vitro, suggesting an enhanced activation response. Moreover, 38% of all juxtavascular cells migrated along a vessel, whereas this was never observed for a nonjuxtavascular cell. These observations identify a mobile subpopulation (10%-30%) of parenchymal microglia that activate rapidly and are preferentially recruited to the surfaces of blood vessels following brain tissue injury. The dynamic and sustained interaction of microglia with brain microvessels may facilitate signaling between injured brain parenchyma and components of the blood-brain barrier or circulating immune cells of the blood in vivo. PMID- 11857682 TI - Human antibodies accelerate the rate of remyelination following lysolecithin induced demyelination in mice. AB - Immunoglobulin-based therapies are becoming increasingly common for the treatment of neurologic and autoimmune diseases in humans. In this study, we demonstrate that systemic administration of either polyclonal human immunoglobulins or specific human monoclonal antibodies can accelerate the rate of CNS remyelination following toxin-induced demyelination. Injection of lysolecithin directly into the spinal cord results in focal demyelinated lesions. In contrast to other murine models of demyelinating disease, the mechanism of demyelination following lysolecithin injection is independent of immune system activation, and chronic inflammation at the site of the lesion is minimal. Administration of polyclonal human IgM (pHIgM) or a serum-derived human monoclonal antibody (sHIgM22) resulted in approximately a twofold increase in remyelinating axons when compared to animals treated with saline or with antibodies that do not promote repair. Both pHIgM and sHIgM22 show strong binding to CNS white matter and oligodendrocytes, while antibodies that did not accelerate remyelination do not. This differential staining pattern suggests that enhanced remyelination may result from direct stimulation of oligodendrocyte remyelination by binding to surface receptors on oligodendrocytes or glial progenitor cells. We propose the use of human polyclonal IgM or specific human monoclonal IgM antibodies as potential therapies to enhance myelin repair following CNS injury and disease. PMID- 11857683 TI - Regulation and differential expression of tau mRNA isoforms as oligodendrocytes mature in vivo: implications for myelination. AB - Oligodendrocytes and neurons derive from the same cell type but develop distinct morphologic and functional properties as they mature in vivo. Both cells express tau protein, a developmentally regulated protein in the central nervous system. The regulation of tau has been investigated extensively in neurons but not in oligodendrocytes, so we studied regulation of tau in oligodendrocytes in vivo. The amino-derived tau isoforms consist of isoforms with zero (A0), one (A1), or two (A2) inserts. We examined the developmental regulation of tau mRNA isoforms at the amino domain by comparing tau expression in oligodendrocytes (OLGs) isolated from 1- and 20-day-old rat brain and in age-matched cortex, which abounds in neurons. In the rat brain, myelination peaks at 20 days. By using semiquantitative RT-PCR, we found that OLGs and cortex from 1-day-old rat brain largely had amino-derived tau isoforms with no insert, whereas OLGs from 20-day old rat brain had similar levels of amino-derived tau isoforms with no insert or with one insert. We also found that 20-day-old OLGs had twofold more tau mRNA levels than younger OLGs. In contrast to OLGs from 20-day-old rat brain, age matched cortex had comparable levels of A0, A1, and A2 tau amino-derived isoforms. Further, younger and older OLGs had a reciprocal pattern of expression of both carboxy-derived tau mRNA isoforms with either three (3R) or four (4R) repeats. In contrast, younger and older cortex expressed either 3R or 4R tau. This study showed an upregulation of tau mRNA and cell-specific tau mRNA isoform expression in OLGs forming myelin. PMID- 11857684 TI - Expression of functional NK-1 receptors in murine microglia. AB - Cells of myeloid origin such as microglia have the potential to contribute significantly to the development of inflammatory responses in the CNS. The ability of the neuropeptide substance P to augment proinflammatory responses by other myeloid cell types such as macrophages and dendritic cells is well recognized. In the present study, we demonstrate the presence of mRNA encoding NK 1 (substance P) receptors in murine microglia cell lines. Importantly, we have utilized specific antibodies developed by our laboratory to detect the expression of the NK-1 receptor protein in murine microglia cell lines by Western blot analysis and flow cytometry. Furthermore, we have investigated the presence of this receptor on primary murine microglia and report the presence of authentic NK 1 receptors as determined by Western blot analysis and flow cytometry. In addition, we demonstrate that NK-1 receptors expressed on microglia are functional as demonstrated by the ability of nanomolar concentrations of substance P to initiate activation of the transcriptional activator, NF-kappaB. Given the weight of evidence supporting the role of substance P--substance P receptor interactions in the initiation of optimal proinflammatory responses by myeloid cells, the demonstration of authentic and functional NK-1 receptors in microglia identifies this neuropeptide as a potentially important contributor to CNS inflammatory responses during disease states. PMID- 11857685 TI - Glucose deprivation decreases nitric oxide production via NADPH depletion in immunostimulated rat primary astrocytes. AB - We have previously reported that the production of nitric oxide (NO) in immunostimulated astrocytes was markedly decreased under glucose-deprived conditions. The present study was undertaken to find the contributing factor(s) for the decreased NO production in glucose-deprived immunostimulated astrocytes. NO production in rat primary astrocytes was stimulated for 24-48 h by cotreatment with lipopolysaccharides (1 microg/ml) and interferon-gamma (100 U/ml). Decreased NO production in immunostimulated astrocytes by glucose deprivation was mimicked by the glycolytic inhibitor 2-deoxyglucose and reversed by addition of pyruvate and lactate. Glucose deprivation did not alter the expression of inducible nitric oxide synthase (iNOS) in immunostimulated astrocytes. Addition of beta-NADPH, but not tetrahydrobiopterine, both of which are essential cofactors for NOS function, completely restored the NO production that was decreased in glucose-deprived immunostimulated astrocytes. Glucose deprivation and immunostimulation synergistically reduced intracellular NADPH level in astrocytes. The results indicate that glucose deprivation decreases NO production in immunostimulated astrocytes by depleting intracellular NADPH, a cofactor of iNOS. PMID- 11857690 TI - Synthetic applications of electrophilic fluorodesilylation. AB - Recently, with the appearance of electrophilic sources of fluorine including the commercially available N-F reagents, the concept of electrophilic fluorodesilylation has emerged as a new strategy to prepare a variety of fluorine containing compounds. This paper highlights how this concept has been applied to the preparation of a series of fluorinated molecules including fluoroaromatic compounds, fluoroalkenes, difluoroamides, difluoroalcohols, difluoroethers and alpha-fluorinated carbonyl derivatives. PMID- 11857691 TI - Syntheses, X-ray crystal structures, and magnetic properties of novel linear M2[II]U[IV] complexes (M=Co, Ni, Cu, Zn). AB - Attempts to prepare heterobimetallic complexes in which 3d and uranium magnetic ions are associated by means of the Schiff bases H(2)L(i) derived from 2 hydroxybenzaldehyde or 2-hydroxy-3-methoxybenzaldehyde were unsuccessful because of ligand transfer reactions between [ML(i)] (M=Co, Ni, Cu) and UCl(4) that led to the mononuclear Schiff base complexes of uranium [UL(i)Cl(2)]. The crystal structure of [UL(3)Cl(2)(py)(2)] [L(3)=N,N'-bis(3-methoxysalicylidene) ethylenediamine; py=pyridine] was determined. The hexadentate Schiff base ligand N,N'-bis(3-hydroxysalicylidene)-2,2-dimethyl-1,3-propanediamine (L) was useful for the synthesis of novel trinuclear complexes of the general formula [[ML(py)](2)U] (M=Co, Ni, Zn) or [[CuL(py)]M'[CuL]] (M'=U, Th, Zr) by reaction of [M(H(2)L)] with [M'(acac)(4)] (acac=MeCOCHCOMe). The crystal structures of the Co(2)U, Ni(2)U, Zn(2)U, Cu(2)U, and Cu(2)Th complexes show that the two ML fragments are orthogonal, being linked to the central actinide ion by the two pairs of oxygen atoms of the Schiff base ligand. In each compound, the UO(8) core exhibits the same dodecahedral geometry, and the three metals are linear. The magnetic study indicated that the two Cu(2+) ions are not coupled in the Cu(2)Zr and Cu(2)Th compounds. The magnetic behavior of the Co(2)U, Ni(2)U, and Cu(2)U complexes was compared with that of the Zn(2)U derivative, in which the paramagnetic 3d ion was replaced with the diamagnetic Zn(2+) ion. A weak antiferromagnetic coupling was observed between the Ni(2+) and the U(4+) ions, while a ferromagnetic interaction was revealed between the Cu(2+) and U(4+) ions. PMID- 11857686 TI - Caveolin-1 expression is maintained in rat and human astroglioma cell lines. AB - Caveolin-1 is the principal structural and functional component of caveolae, a plasmalemmal compartment that has been proposed to sequester lipid and protein components that participate in transmembrane signal transduction processes. Multiple studies reveal a reduction in the expression level of caveolin-1 mRNA and protein in many carcinomas as well as transformed cells. The human caveolin-1 gene is localized to a suspected tumor suppressor locus (7q31.1). Collectively, these data have been taken to imply that caveolin-1 may function in a tumor suppressor capacity. To determine if a reduction in the expression level of caveolin-1 mRNA and protein accompanied the transformation of astrocytes, we undertook studies of two transformed rat astroglial cell lines, C6 and DI TNC(1), as well as several cell lines derived from human glioblastoma tumors: T98G, U87MG, U118MG, U138MG, and U373MG. Ultrastructural, immunolocalization, immunoblot, and Northern blot analyses demonstrated that caveolin-1 message and protein were expressed in all rat and human glioma cells. The localization pattern, buoyant density, and detergent-insolubility property of caveolin-1 protein were indistinguishable from that determined for nontransformed type 1 astrocytes in culture. Nucleotide sequence analyses of caveolin-1 cDNAs indicate that mutations are not present in the caveolin-1 sequence in any of the glioma cell types. Taken together with previous analyses, these data indicate that, at least for astrocytes, the process of transformation in and of itself is not solely sufficient to reduce the level of caveolin-1 expression, and that caveolin 1 expression in and of itself is not solely sufficient to prevent the acquisition of a transformed phenotype. PMID- 11857692 TI - Effects of structural factors on the pi-dimerization and/or disproportionation of the cation radical of extended TTF containing thiophene-based pi-conjugated spacers. AB - The electrochemical and chemical oxidation of extended TTF 4 and 5 are analysed by cyclic voltammetry, Visible/NIR and ESR spectroscopies, and the X-ray structures of the new salts 5 x BF(4)(CH(2)Cl(2)) and 4 x ClO(4)(THF)(1/2) are presented. The effects of structural factors on the pi-dimerization or the disproportionation reaction of the cation radical are shown. The oxidation of compound 4 presents the successive formation of stable cation radical and dication species both in dichloromethane (DCM) and in a CH(3)CN/THF mixture. In contrast, for compound 5, the stability of the oxidation states strongly depends on the nature of the solvent. In DCM, the oxidation of 5 proceeds by two close one-electron transfers while in CH(3)CN/THF the dication is directly formed via a two-electron process. The X-ray structures of the two salts reveal the formation of pi-dimers of cation radical. While the dimer (5(2))(2+) is due mainly to pi-pi interactions between the conjugating spacer, the multiplication of the sulfur atoms in compound 4 contributes to stabilize the dimer by the combined effects of S-S and pi-pi interactions. Visible/NIR and ESR experiments confirm the higher tendency of 4(+)(.) to dimerize with the occurrence of dimer and monomer in solution, while for 5(+)(.) only the monomer is detected in DCM. On the other hand, by dissolution of 5 x BF(4)(CH(2)Cl(2)) in CH(3)CN, only the neutral and the dicationic states of compounds 5 are observed owing to the disproportionation reaction. PMID- 11857693 TI - How to synthesize macrocycles efficiently by using virtual combinatorial libraries. AB - The selection of different diimines 4 a-c by alkaline earth ions from a virtual combinatorial library (VCL) is described. The products were stabilized by reduction to the diamines 6 a-c; this allowed easy analysis. The library can be directed toward different target molecules 6 a-c upon addition of alkaline earth ions with different radii. Competition experiments show the possibility of synthesizing the macrocycles 6 a, 6 b, and 6 c simultaneously when using Mg(2+), Ca(2+), and Ba(2+) as template ions. The scope of this thermodynamically controlled, reversible approach for macrocycle syntheses is illustrated. PMID- 11857694 TI - The trifluoromethoxy group: a long-range electron-withdrawing substituent. AB - Judged by its capacity to promote a hydrogen/metal permutation at an ortho position, the trifluoromethoxy group is superior to both the methoxy and trifluoromethyl groups. Moreover, like CF(3) and unlike OCH(3), OCF(3) exerts a long-range effect that still considerably lowers the basicity of arylmetal compounds when located in a more remote meta or even para position. As a consequence, 4-(trifluoromethoxy)anisole is deprotonated by sec-butyllithium mainly, and by tert-butyllithium exclusively, at a position adjacent to the OCH(3) group rather than next to the strongly electron-withdrawing CF(3)O group. 1,3-Benzodioxole undergoes ortho lithiation only six times faster than anisole, whereas 2,2-difluoro-1,3-benzodioxole reacts about 5000 times faster, as evidenced by competition experiments. The structure and distance dependence of substituent effects can be rationalized by assuming superposing sigma- and pi polarizing interactions. PMID- 11857695 TI - Transfer hydrogenation processes to mu(3)-alkylidyne groups on the organotitanium oxide [Ti(3)Cp*O(3)]. AB - The photochemical treatment of mu(3)-alkylidyne complexes [[TiCp*(mu-O)](3)(mu(3) CR)] (R=H (1), Me (2), Cp*=eta(5)-C(5)Me(5)) with the amines (2,6 Me(2)C(6)H(3))NH(2), Et(2)NH, and Ph(2)NH and the imine Ph(2)C=NH leads to the partial hydrogenation of the alkylidyne moiety that is supported on the organometallic oxide, [Ti(3)Cp*O(3)], and the formation of new oxoderivatives [[TiCp*(3)(mu-CHR)(R'NR")] (R"=2,6-Me(2)C(6)H(3), R'=H, R=H (3), Me (4); R'=R"=Et, R=H (5), Me (6); R'=R"=Ph, R=H (7), Me (8)) and [[TiCp*(mu-O)](3)(mu CHR)(N=CPh(2))] (R=H (9), R=Me (10)), respectively. A sequential transfer hydrogenation process occurs when complex 1 is treated with tBuNH(2), which initially gives the mu-methylene [[TiCp*(mu-O)](3)(mu-CH(2))(HNtBu)] (11) complex and finally, the alkyl derivative [[TiCp*(mu-O)](3)(mu-NtBu)Me] (12). Furthermore, irradiation of solutions of the mu(3)-alkylidyne complexes 1 or 2 in the presence of diamines o-C(6)H(4)(NH(2))(2) and H(2)NCH(2)CH(2)NH(2) (en) affords [[TiCp*(mu-O)](3)(mu(3)-eta(2)-NC(6)H(4)NH)] (13) and [[TiCp*(mu O)](3)(mu(3)-eta(2)-NC(2)H(4)NH)] (14) by either methane or ethane elimination, respectively. In the reaction of 1 with en, an intermediate complex [[TiCp*(mu O)](3)(mu-CH(2))(NHCH(2)CH(2)NH(2))] (15) is detected by (1)H NMR spectroscopy. Thermal treatment of the complexes 4-10 quantitatively regenerates the starting mu(3)-alkylidyne compounds and the amine R'(2)NH or the imine Ph(2)C=NH; however, heating of solutions of 3 or 4 in [D(6)]benzene or a equimolecular mixture of both at 170 degrees C produces methane, ethane, or both, and the complex [[TiCp*(mu-O)](3)[mu(3)-eta(2)-NC(6)H(3)(Me)CH(2)]] (16). The molecular structure of 8 has been established by single-crystal X-ray analysis. PMID- 11857696 TI - Dendritic galactosides based on a beta-cyclodextrin core for the construction of site-specific molecular delivery systems: synthesis and molecular recognition studies. AB - In order to evaluate the ability of multivalent glycosides based on a beta cyclodextrin core as site-specific molecular carriers, a study on both the inclusion complexation behaviour and lectin binding affinity of branched and hyperbranched beta-cyclodextrins is presented. A series of cluster galactosides constructed on beta-cyclodextrin scaffolds containing seven 1-thio-beta-lactose or beta-lactosylamine bound to the macrocyclic core through different spacer arms were synthesised. In addition, the first synthesis of three first-order dendrimers based on a beta-cyclodextrin core containing fourteen 1-thio-beta-D galactose, 1-thio-beta-lactose and 1-thio-beta-melibiose residues was performed. Calorimetric titrations performed at 25 degrees C in buffered aqueous solution (pH 7.4) gave the affinity constants and the thermodynamic parameters for the complex formation of these beta-cyclodextrin derivatives with guests sodium 8 anilino-1-naphthalenesulfonate (ANS) and 2-naphthalenesulfonate, and lectin from peanut (Arachis hypogaea) (PNA). The persubstitution of the primary face of the beta-cyclodextrin with saccharides led to a slight increase of the binding constant values for the inclusion complexation with ANS relative to the native beta-cyclodextrin. However, the increase of the steric congestion due to the presence of the saccharide residues on the narrow rim of the beta-cyclodextrin may cause a decrease of the binding ability as shown for sodium 2 naphthalenesulfonate. The spacer arms are not passive elements and influence the host binding ability according to their chemical nature. PNA forms soluble cross linked complexes with cluster galactosides and lactosides scaffolded on beta cyclodextrin but not with cluster galactopyranosylamines or melibiose. Both, perbranched and hyperbranched beta-cyclodextrins, form stronger complexes with PNA than the monomeric analogues. However, the use of hyperbranched CDs does not contribute to the improvement of the complex stability relative to heptakis glycocyclodextrin derivatives. Finally, a titration experiment with PNA and a complex formed by a heptakis lactose beta-cyclodextrin derivative with sodium 2 naphthalenesulfonate showed the formation of a soluble cross-linked complex with stronger affinity constant and higher stoichiometry than those observed for the complex formation of PNA with the same heptakis-lactose beta-cyclodextrin derivative, suggesting the formation of a three component complex. PMID- 11857697 TI - The first enantiomerically pure [n]triangulanes and analogues: sigma-[n]helicenes with remarkable features. AB - (M)-(-)- and (P)-(+)-Trispiro[2.0.0.2.1.1]nonanes [(M)- and (P)-3] as well as (M) (-)- and (P)-(+)-tetraspiro[2.0.0.0.2.1.1.1]undecanes [(M)- and (P)-4] enantiomerically pure unbranched [4]- and [5]triangulanes-have been prepared starting from racemic bicyclopropylidenecarboxylic [(1RS)-12] and exo dispiro[2.0.2.1]heptane-1-carboxylic [(1RS,3SR)-13] acids. The optical resolutions of rac-12 and rac-13 furnished enantiomerically pure acids (S)-(+) 12, (R)-(-)-12, (1R,3S)-(-)-13, and (1S,3R)-(+)-13. The ethyl ester (R)-25 of the acid (R)-(-)-12 was cyclopropanated to give carboxylates (1R,3R)-26 and (1R,3S) 26. The ester (1R,3S)-26 and acids (1R,3S)-13 and (1S,3R)-13 were converted into enantiomerically pure methylene[3]triangulanes (S)-(-)- and (R)-(+)-28. An alternative approach consisted of an enzymatic deracemization of endo-[(1SR,3SR) dispiro[2.0.2.1]heptyl]methanol (rac-20) or anti-[(1SR,3RS)-4 methylenespiropentyl]methanol (rac-18). This afforded (S)-(-)- and (R)-(+)-28 (starting from rac-20), as well as enantiomerically pure (M)-(-)- and (P)-(+)-1,4 dimethylenespiropentanes [(M)- and (P)-23] starting from rac-18. The methylenetriangulanes (S)-(-)- and (R)-(+)-28 were cyclopropanated furnishing (M) and (P)-3. The rhodium-catalyzed cycloaddition of ethyl diazoacetate onto (S)-( )- and (R)-(+)-28 yielded four diastereomeric ethyl trispiro[2.0.0.2.1.1]nonane-1 carboxylates in approximately equal proportions. The enantiomerically pure esters (1R,3S,4S)- and (1S,3R,4R)-30 were isolated by careful distillation and then transformed into [5]triangulanes (M)- and (P)-4 using the same sequence of reactions as applied for (M)- and (P)-3. The structures of the key intermediates (R)-12 and rac-31 were confirmed by X-ray analyses. Although [4]- and [5]triangulanes have no chromophore which would lead to any significant absorption above 200 nm, they have remarkably high specific rotations even at 589 nm with [alpha](20)D=-192.7 [(M)-3, c=1.18, CHCl(3))] or +373.0 [(P)-4, c=1.18, CHCl(3))]. This remarkable optical rotatation is in line with their helical arrangement of sigma bonds, as confirmed by a full valence space single excitation configuration interaction treatment (SCI) in conjunction with DFT computations at the B3LYP/TZVP//B3LYP/6-31+G(d,p) level of theory which reproduce the ORD very well. Thus, it is appropriate to call the helically shaped unbranched [n]triangulanes the "sigma-[n]helicenes", representing the sigma-bond analogues of the aromatic [n]helicenes. PMID- 11857698 TI - Synthesis of a new class of chiral 1,5-diphosphanylferrocene ligands and their use in enantioselective hydrogenation. AB - A new family of ferrocenylphosphane ligands has been prepared. Their flexible synthesis allows many structural modifications. The asymmetric induction of these ligands was examined in the hydrogenation of functionalized C=C, C=O, and C=N bonds. The enantioselectivity of the reaction was strongly dependent on the substituent R at the position alpha to the ferrocene moiety. In many cases, both enantiomeric beta-hydroxyesters of the reduction product can be obtained by simply replacing a dimethylamino group in the ligand with a methyl group. PMID- 11857699 TI - Synthesis of (-)-Vulcanolide by enantioselective protonation. AB - Two efficient enantioselective syntheses of the more active (S,S)-enantiomer of the powerful musk odorant Vulcanolide are described. In both syntheses, the key step is an enantioselective protonation of a ketone enolate. A third enantioselective protonation, of a thiol ester enolate, was applied for the determination of the absolute configuration of Vulcanolide by comparison with a known compound. PMID- 11857700 TI - Water-assisted alkaline hydrolysis of monobactams: a theoretical study. AB - A theoretical study of the water-assisted alkaline hydrolysis of 2-azetidinone, 3 formylamino-2-azetidinone and 3-formylamino-2-azetidine-1-sulfonate ion is carried out at the B3LYP/6-31+G* level. The effect of bulk solvent is taken into account using the PCM solvation model while specific solvent effects are represented by the inclusion of an ancillary water molecule along the reaction profile. The calculated free energy barriers in solution are in reasonable agreement with experimental values. The observed substituent effects due to the presence of the 3-formylamino and the SO(3) groups attached to the beta-lactam ring are crucial factors determining the hydrolysis of monobactam antibiotics. PMID- 11857701 TI - The nature of the indenyl effect. AB - The eta(5)-to-eta(3) coordination shift of cyclopentadienyl (Cp=C(5)H(5)(-)) and indenyl (Ind=C(9)H(7)(-)) ligands in molybdenocene complexes, [(eta(5) Cp')(eta(5)-Cp)Mo(CO)(2)](2+) (Cp'=Cp or Ind), driven by a two-electron reduction of those species, was studied and compared by means of molecular orbital calculations (B3LYP HF/DFT hybrid functional, DZP basis sets). The results obtained, in terms of optimized geometries, relative energies, and bond analysis parameters, compare well with the experimental data, and verify the well-known indenyl effect, that is, a significantly more facile eta(5)-to-eta(3) rearrangement for the indenyl ligand when compared to cyclopentadienyl. However, the study of the folding of free Cp and Ind, combined with the (eta(5/3)-Cp')-M bond analysis, shows that the observed difference is not the result of an intrinsic characteristic of the indenyl ligand, such as the traditionally accepted aromaticity gain in the benzene ring formed in eta(3)-Ind complexes. Instead, it is directly related to the Cp'-M bond strength. While the difference in the energy required to fold the two free ligands is negligible (< or =1 kcal mol(-1) for folding angles up to 20 degrees), the (eta(5)-Cp)-M bond is stronger than that of (eta(5)-Ind)-M; however, the opposite situation is found for the eta(3) coordination mode. The net result, for Cp'=Ind, is a destabilization of the eta(5) complexes and a stabilization of the eta(3) intermediates or transition states yielding smaller activation energies and faster reaction rates for processes in which that is the rate-determining step. PMID- 11857702 TI - Self-assembly of molecular dumbbells into organized bundles with tunable size. AB - Dumbbell-shaped molecules consisting of three biphenyls connected through vinyl linkages as a conjugated rod segment and aliphatic polyether dendritic wedges with different cross-sections (i.e., dibranch (1), tetrabranch (2) and hexabranch (3)) were synthesized and characterized. The molecular dumbbells self-assemble into discrete bundles that organize into three-dimensional superlattices. Molecule 1, based on a dibranched dendritic wedge, organizes into primitive monoclinic-crystalline and body-centered, tetragonal liquid crystalline structures, while molecules 2 and 3, based on tetra- and hexabranched dendritic wedges, respectively, form only body-centered, tetragonal liquid crystalline structures. X-ray diffraction experiments and density measurements showed that the rod-bundle cross-sectional area decreases with increasing cross-section of the dendritic wedges. The influences of supramolecular structure on the bulk state optical properties were investigated by measuring the UV/Vis absorption and steady state fluorescence spectroscopies. As the cross-section of the dendritic wedge of the molecule increases, the absorption and emission maxima shift to higher energy. This can be attributed to a quantum size effect of the three dimensionally confined nanostructure. PMID- 11857703 TI - A practical route to enantiopure, highly functionalized seven-membered carbocycles and tetrahydrofurans: concise synthesis of (+)-nemorensic Acid. AB - Highly diastereoselective thermal [5C+2C] intramolecular pyrone-alkene cycloadditions can be achieved by introducing a homochiral p-tolylsulfinyl group at a suitable position of the alkene. The resulting adducts can be readily desulfinylated to give optically active 8-oxabicyclo[3.2.1]octane derivatives. Interestingly, switching from a sulfinyl to a sulfonimidoyl group allows one to reverse the direction of the diastereofacial selectivity and thereby produces oxa bridged carbocyclic systems enantiomeric to those obtained from the sulfinyl precursors. Cleavage of the oxa-bridge on the desulfurated adducts yields highly functionalized seven-membered carbocyclic derivatives in enantiopure form. Alternative cleavage of the seven-membered carbocycle provides enantiomerically enriched tetrahydrofurans. We have exploited this reaction pathway for the synthesis of the naturally occurring enantiomer of nemorensic acid. PMID- 11857704 TI - Anchor dependency for non-glycerol based cationic lipofectins: mixed bag of regular and anomalous transfection profiles. AB - Although detailed structure-activity, physicochemical and biophysical investigations in probing the anchor influence in liposomal gene delivery have been reported for glycerol-based transfection lipids, the corresponding investigation for non-glycerol based simple monocationic transfection lipids have not yet been undertaken. Towards this end, herein, we delineate our structure activity and physicochemical approach in deciphering the anchor dependency in liposomal gene delivery using fifteen new structural analogues (lipids 1-15) of recently reported non-glycerol based monocationic transfection lipids. The C(14) analogues in both series 1 (lipids 1-6) and series 2 (lipids 7-15) showed maximum efficiency in transfecting COS-1 and CHO cells. However, the C(12) analogue of the ether series (lipid 3) exhibited a seemingly anomalous behavior compared with its transfection efficient C(10) and C(14) analogues (lipids 2 and 4) in being completely inefficient to transfect both COS-1 and CHO cells. The present structure-activity investigation also convincingly demonstrates that enhancement of transfection efficiencies through incorporation of membrane reorganizing unsaturation elements in the hydrophobic anchor of cationic lipids is not universal but cell dependent. The strength of the interaction of lipids 1-15 with DNA was assessed by their ability to exclude ethidium bromide bound to the DNA. Cationic lipids with long hydrophobic tails were found, in general, to be efficient in excluding EtBr from DNA. Gel to liquid crystalline transition temperatures of the lipids was measured by fluorescence anisotropy measurement technique. In general (lipid 2 being an exception), transfection efficient lipids were found to have their mid transition temperatures at or below physiological temperatures (37 degrees C). PMID- 11857705 TI - Stereocontrolled synthesis of angularly fused tricyclic ring systems by means of 1-metalla-1,3,5-hexatrienes (M=Cr, W). AB - An efficient pathway for the stereocontrolled synthesis of functionalized, angularly fused tricyclic ring systems from readily available (1-alkynyl)carbene complexes [(OC)(5)M=C(OEt)C(triple bond)CR] (M=Cr, W; R=Ph, c-C(6)H(9)) is described. The synthesis involves the formation of a 1-metalla-1,3,5-hexatriene from the (1-alkynyl)carbene tungsten complex [(OC)(5)W=C(OEt)C(triple bond)C-c C(6)H(9)] and a secondary amine, and its thermally induced pi-cyclization to a tetrahydroindene, which undergoes a spontaneous isomerization to another tetrahydroindene. Condensation of these tetrahydroindenes with pyran-2-ylidene complexes derived from (1-alkynyl)carbene complexes [(OC)(5)M=C(OEt)C(triple bond)CPh] (M=Cr, W) proceeds smoothly giving angularly fused tricyclic ring systems, rearrangement of which may generate spiro(cyclopentane-1,1-indanes) as side products. The synthesis is highly versatile and can be applied to the formation of various ring systems, such as steroid-type ring skeletons. PMID- 11857707 TI - A cuboctahedral supramolecular capsule by 4:4 self-assembly of Tris(Zn(II) cyclen) and trianionic trithiocyanurate in aqueous solution at neutral pH (cyclen=1,4,7,10-tetraazacyclododecane). AB - A 1:1 mixture of a tris(Zn(II)-cyclen) (1: [Zn(3)L(1)], L(1)=1,3,5-tris(1,4,7,10 tetraazacyclododecan-1-ylmethyl)benzene) and trithiocyanuric acid (TCA) yielded a 4:4 self-assembly complex [(Zn(3)L(1))(4)-(TCA(3-))(4)] (6) through the formation of Zn(II)-S(-) coordination bonds and hydrogen bonds between 1,3,5-triazine N and cyclen NH (cyclen=1,4,7,10-tetraazacyclododecane); the supramolecular capsule structure was revealed by X-ray crystal structure analysis. The capsule exterior represents a twisted cuboctahedral framework containing a nanoscale truncated tetrahedral cavity. The crystal data: formula C(144)H(308)N(72)O(58)S(12)Zn(12) (6[NO(3)](12) x 22 H(2)O), M(r)=5145.75, cubic, space group F432 (No. 209), a=39.182(1) A, V=60153(3) A (3), Z=8, R=0.100, R(w)=0.259. Lipophilic organic molecules with the matching sizes, for example, ([D(4)]-2,2,3,3)-3 (trimethylsilyl)propionic acid (TSP), 1-adamantanecarboxylic acid, 2,4 dinitrophenol (2,4-DNP), adamantane (ADM), or the tetra-n-propylammonium (TPA) cation, are encapsulated in the inner cavity, as revealed by remarkable upfield shifts of the (1)H NMR signals of these guest molecules. The encapsulation of ADM was confirmed by X-ray crystal structure analysis. Crystal data of the ADM encapsulating complex: formula C(154)H(334)N(72)O(63)S(12)Zn(12) (6 ADM[NO(3)](12) x 27 H(2)O), M(r)=5372.06, cubic, space group F432 (No. 209), a=39.061(1) A, V=59599(3) A(3), Z=8, R=0.103, R(w)=0.263. The 4:4 self-assembly was stabilized by incorporation of one of these guest molecules. The apparent 4:4 self-assembly constants for 6 in the presence of an excess amount of a guest TPA, log K(app) (K(app)=[6-TPA]/[1](4)[TCA](4)) (M(-7))), were determined to be 34.0+/ 2.0 and 35.5+/-3.0 by potentiometric pH and UV spectrophotometric titrations, respectively. An apparent encapsulation constant for 2,4-DNP, log K(enc) (K(enc)=[6-2,4-DNP]/[6][2,4-DNP] (M(-1))), was 6.0+/-0.1 at pH 7.0 (50 mM HEPES with I=0.1 (NaNO(3))), as determined by UV titrations. The lipophilicity of the inner cavity was close to that of 2-propanol, as a quantum yield (phi) of 0.24+/ 0.1 for the fluorescent emission of 7-diethylaminocoumarin-1-carboxylic acid (20 microM) in the capsule was close to the phi of 0.22 found for 2-propanol. Encapsulation properties of the present Zn(II)-containing cage have been compared with those of cyclodextrins and Fujita's Pd(II)-containing supramolecular cage. The exterior chirality of the 4:4 complex was controlled from within by an encapsulated chiral guest molecule, 2,10-camphorsultam, as indicated by Cotton effects in the circular dichroism spectra. PMID- 11857708 TI - The interaction of water and dibromine in the gas phase: an investigation of the complex H(2)O...Br(2) by rotational spectroscopy and ab initio calculations. AB - The ground-state rotational spectra of eight isotopomers of a complex formed by water and dibromine in the gas phase were observed by pulsed-jet, Fourier transform microwave spectroscopy. The spectroscopic constants B(0), C(0), delta(J), delta(JK), chi(aa)(Br(x)) (x=i for inner, o for outer), [chi(bb)(Br(x)) chi(cc)(Br(x))] and M(bb)(Br(x)) were determined for H(2)O...(79)Br(79)Br, H(2)O...(81)Br(79)Br, H(2)O...(79)Br(81)Br, H(2)O...(81)Br(81)Br, D(2)O...(79)Br(81)Br and D(2)O...(81)Br(81)Br. For the isotopomers HDO...(79)Br(81)Br and HDO...(81)Br(81)Br, only (B(0) + C(0))/2, delta(J), the chi(aa)(Br(x)) and M(bb)(Br(x)) were determinable. The spectroscopic constants were interpreted on the basis of several models of the complex to give information about its geometry, binding strength and the extent of electronic rearrangement on complex formation. The molecule H(2)O...Br(2) has C(s) symmetry with a pyramidal configuration at O. The zero-point effective quantities r(O...Br(i))=2.8506(1) A and phi(0)=46.8(1), where phi is the angle between the C(2) axis of H(2)O and the O...Br-Br internuclear axis, were obtained under the assumption of monomer geometries unchanged by complexation. Ab initio calculations, carried out at the aug-cc-pVDZ/MP2 level of theory, gave the equilibrium values r(e)(O...Br(i))=2.7908 A and phi(e)=45.7 degrees and confirmed the collinearity of the O...Br-Br nuclei. The potential energy function V(phi), also determined ab initio, showed that the wavenumber required for inversion of the configuration at O in the zero-point state is only 9 cm(-1). By interpreting the Br nuclear quadrupole coupling constants, the fractions delta(O- >Br(i))=0.004(5) and delta (Br(i)-->Br(o))=0.050(2) of an electron were determined to be transferred from O to Br(i) and Br(i) to Br(o), respectively, when the complex is formed. The complex is relatively weak, as indicated by the small value k(sigma)=9.8(2) N m(-1) of the intermolecular stretching force constant obtained from delta(J). A comparison of the properties, similarly determined, of H(2)O...F(2), H(2)O...Cl(2), H(2)O...Br(2), H(2)O...BrCl, H(2)O...ClF and H(2)O...ICl is presented. PMID- 11857706 TI - Efficient and stereoselective synthesis of bicyclo[3.2.1]octan-8-ones: synthesis and palladium-catalyzed isomerization of functionalized 2-vinyl-2,3,3a,4,5,6 hexahydro-2,3-benzofurans. AB - A new C,O-cyclodialkylation of dilithiated cyclic beta-keto esters and beta-keto sulfones with 1,4-dibromo-2-butene is reported which results in regio- and diastereoselective formation of 2-vinyl-2,3,3a,4,5,6-hexahydro-2,3-benzofurans. The products could be efficiently transformed into functionalized bicyclo[3.2.1]octan-8-ones by a palladium-catalyzed rearrangement reaction. In case of sulfone derivatives, this rearrangement proceeds with high stereospecifity to give exclusively the endo-configured diastereomers. The bicyclo[3.2.1]octane skeleton is present in a large number of pharmacologically important natural products. PMID- 11857709 TI - Supramolecular assemblies of a bis(terpyridine) ligand and of its [2x2] grid-type Zn(II) and Co(II) complexes on highly ordered pyrolytic graphite. AB - The synthesis and characterisation of bis(terpyridine)-derived ligands which are capable of forming [2x2] grid-like complexes is presented. Additional pyridine substituents on these ligands do not interfere with the complexation process. Adsorbing one of the pure ligands on highly ordered pyrolytic graphite (HOPG) shows a highly ordered structure stabilised by additional weak intermolecular C H...N hydrogen bonds partially through the extra-pyridines as could be shown by scanning tunneling microscopy (STM) investigations. Similar adsorption experiments with one of the corresponding [2x2] Co(II) grid-type complexes on graphite, led also to a well-organised structure with interdigitation of the extra-pyridine moieties. PMID- 11857710 TI - (NEt(4))(2)[Fe(CN)(2)(CO)('S(3)')]: an iron thiolate complex modeling the [Fe(CN)(2)(CO)(S-Cys)(2)] site of [NiFe] hydrogenase centers. AB - In the search for complexes modeling the [Fe(CN)(2)(CO)(cysteinate)(2)] cores of the active centers of [NiFe] hydrogenases, the complex (NEt(4))(2)[Fe(CN)(2)(CO)('S(3)')] (4) was found ('S(3)'(2-)=bis(2 mercaptophenyl)sulfide(2-)). Starting complex for the synthesis of 4 was [Fe(CO)(2)('S(3)')](2) (1). Complex 1 formed from [Fe(CO)(3)(PhCH=CHCOMe)] and neutral 'S(3)'-H(2). Reactions of 1 with PCy(3) or DPPE (1,2 bis(diphenylphosphino)ethane) yielded diastereoselectively [Fe(CO)(2)(PCy(3))('S(3)')] (2) and [Fe(CO)(dppe)('S(3)')] (3). The diastereoselective formation of 2 and 3 is rationalized by the trans influence of the 'S(3)'(2-) thiolate and thioether S atoms which act as pi donors and pi acceptors, respectively. The trans influence of the 'S(3)'(2-) sulfur donors also rationalizes the diastereoselective formation of the C(1) symmetrical anion of 4, when 1 is treated with four equivalents of NEt(4)CN. The molecular structures of 1, 3 x 0.5 C(7)H(8), and (AsPh(4))(2)[Fe(CN)(2)(CO)('S(3)')] x acetone (4 a x C(3)H(6)O) were determined by X-ray structure analyses. Complex 4 is the first complex that models the unusual 2:1 cyano/carbonyl and dithiolate coordination of the [NiFe] hydrogenase iron site. Complex 4 can be reversibly oxidized electrochemically; chemical oxidation of 4 by [Fe(Cp)(2)PF(6)], however, led to loss of the CO ligand and yielded only products, which could not be characterized. When dissolved in solvents of increasing proton activity (from CH(3)CN to buffered H(2)O), complex 4 exhibits drastic nu(CO) blue shifts of up to 44 cm(-1), and relatively small nu(CN) red shifts of approximately 10 cm(-1). The nu(CO) frequency of 4 in H(2)O (1973 cm(-1)) is higher than that of any hydrogenase state (1952 cm(-1)). In addition, the nu(CO) frequency shift of 4 in various solvents is larger than that of [NiFe] hydrogenase in its most reduced or oxidized state. These results demonstrate that complexes modeling properly the nu(CO) frequencies of [NiFe] hydrogenase probably need a [Ni(thiolate)(2)] unit. The results also demonstrate that the nu(CO) frequency of [Fe(CN)(2)(CO)(thiolate)(2)] complexes is more significantly shifted by changing the solvent than the nu(CO) frequency of [NiFe] hydrogenases by coupled-proton and electron-transfer reactions. The "iron-wheel" complex [Fe(6)[Fe('S(3)')(2)](6)] (6) resulting as a minor by-product from the recrystallization of 2 in boiling toluene could be characterized by X-ray structure analysis. PMID- 11857711 TI - Strain and reactivity: electrophilic addition of bromine and tribromide salts to cyclic allenes. AB - The kinetics and the products of the bromination of several cyclic allenes, from C(9) to C(13) (1 a-e), with tetrabutylammonium tribromide (TBAT) and Br(2) have been investigated in 1,2-dichloroethane (DCE) and methanol. The first product of the interaction between the allene and Br(2) is a 1:1 pi complex. The stability constant of this complex, determined at 25 degrees C for allene 1 a, is 7.4 M( 1). The comparison of this value with those reported for several alkenes and alkynes further support the hypothesis of the existence of sizeable structural effects on the stability of these complexes. The negative values of the apparent activation energy for the reaction of allenes 1 a-e with Br(2) in DCE demonstrate the involvement of these complexes as essential intermediates along the reaction coordinate. Different stereochemical behavior was observed in the bromine addition on going from the strained 1,2-cyclononadiene to the larger compounds. Furthermore, a solvent-dependent stereochemistry has been observed for each compound. The kinetic and product distribution data have been interpreted in terms of the influence of the strain on the nature of the intermediate and by considering the competition between pre-association and ion-pair pathways on going from aprotic to nuclophilic solvents or when nucleophilic bromide ions are added. Ab initio (MP2/6-311+G**) and density functional (B3LYP/6-311+G**) computations of 1:1 Br(2) complexes showed that the association energies of allene x Br(2) and ethene x Br(2) complexes are nearly the same but are greater than that of acetylene x Br(2) complexes. Allene x 2 Br(2) complexes are more stable than their ethene x 2 Br(2) counterparts. Br(2) x allene x Br(2) structures, in which the bromine molecules interact either with a single allene double bond or individually with both double bonds, are not preferred significantly over alternatives with Br(2)...Br(2) interactions. As a result of the entropy, the association of bromine with unsaturated hydrocarbons is usually unfavorable in the gas phase (except at extremely low temperatures); complexes are observed in solution (under ambient conditions), since the entropy loss is reduced as a result of restricted translation and rotation and possible association to the solvent. The 1,2-cycloheptadiene x Br(2) > 1,2-cyclononadiene x Br(2) > 1,3-dimethylallene x Br(2) association energies increase with ring strain. PMID- 11857712 TI - T(h)-symmetrical hexakisadducts of C(60) with a densely packed pi-donor shell can act as energy- or electron-transducing systems. AB - For the first time several T(h)-symmetrical hexakisadducts of C(60) bearing up to six electro- and photoactive o-phenylene diamine or 9,10-dialkoxyanthracene moieties were synthesized and subjected to photoinduced electron/energy-transfer studies. Both donors form a densely packed pi-donor shell surrounding the fullerene core. In these novel core-shell ensembles (7 and 19), either an efficient energy transfer from the dialkoxyanthracene periphery, or an electron transfer from the o-phenylene diamine periphery transduces the flow of excited state energy or electrons, respectively, to the fullerene moiety, which resides in the central core. Due to the relatively high reduction potential of the fullerene core, which is anodically shifted by approximately equal to 0.7 V, compared with that of pristine C(60), the outcome of these intramolecular reactions depends mainly on the donor ability of the peripheral system. Interestingly, the charge-separated state in the o-phenylene diamine heptad (7; tau=2380 ns in benzonitrile) is stabilized by a factor of 20 relative to the corresponding o-phenylene diamine dyad (6; tau=120 ns in benzonitrile), an effect that points unequivocally to the optimized storage of charges in this highly functionalized fullerene ensemble. PMID- 11857714 TI - Arsenosugar identification in seaweed extracts using high-performance liquid chromatography/electrospray ion trap mass spectrometry. AB - The development of analytical techniques suitable for providing structural information on a wide range of elemental species is a growing necessity. For arsenic speciation a variety of mass spectrometric techniques, mainly inductively coupled plasma mass spectrometry (ICP-MS) and electrospray tandem mass spectrometry (ES-MS/MS) coupled on-line with high-performance liquid chromatography (HPLC), are in use. In this paper we report the identification of arsenic species present in samples of marine origin (seaweed extracts) using ES ion trap mass spectrometry (IT) multistage mass spectrometry (MS(n)). Both reversed-phase and anion-exchange HPLC have been coupled on-line to ES-ITMS. Product ion scans with multiple stages of tandem MS (MS(n); n=2-4) were used to acquire diagnostic data for each arsenosugar. The spectra contain structurally characteristic fragment ions for each of the arsenosugars examined. In addition it was observed that upon successive stages of collision-induced dissociation (CID) a common product ion (m/z 237) was formed from all four arsenosugars examined. This product ion has the potential to be used as an indicator for the presence of dimethylated arsenosugars (dimethylarsinoylribosides). The HPLC/ES ITMS(n) method developed allows the sensitive identification of arsenosugars present in crude seaweed extracts without the need for extended sample preparation. In fact, sample preparation requirements are identical to those typically employed for HPLC/ICP-MS analysis. Additionally, the resulting product ions are structurally diagnostic of the arsenosugars examined, and tandem mass spectra are reproducible and correspond well to those obtained using other low energy CID techniques. As a result, the HPLC/ES-ITMS(n) approach minimises the potential for arsenic species misidentification and has great potential as a means of overcoming the need for characterised standards. PMID- 11857715 TI - Enantiomeric separation and quantification of fluoxetine (Prozac) in human plasma by liquid chromatography/tandem mass spectrometry using liquid-liquid extraction in 96-well plate format. AB - Fluoxetine (Prozac) is currently one of the widely prescribed selective serotonin reuptake inhibitors (SSRIs) for the treatment of depression. A high-throughput sample preparation procedure using liquid-liquid extraction (LLE) in a 96-well plate format in conjunction with liquid chromatography/tandem mass spectrometry (LC/MS/MS) was developed and validated for quantification of fluoxetine enantiomers in human plasma. After addition of internal standard and ammonium hydroxide, samples were extracted with ethyl acetate. The organic extract was evaporated to dryness and reconstituted in methanol. Where possible, sample transfer and LLE steps were automated using a Tomtec Quadra 96 workstation. Adequate separation of fluoxetine enantiomeric pairs (resolution of 1.17) was achieved on a vancomycin column eluted with methanol containing 0.075% (by weight) ammonium trifluoroacetate. A triple quadrupole mass spectrometer, operated in the multiple reaction monitoring mode at m/z 310-->44 for fluoxetine enantiomeric pairs and m/z 287-->241 for oxazepam (internal standard), was used. Analysis was performed in the positive ion mode using atmospheric pressure chemical ionization (APCI). The standard curve range was 2.0-1000 ng/mL for each fluoxetine enantiomer. The intra- and inter-day precision and accuracy of the quality control (QC) samples were <12.5% (CV) and <13.6% (CV), respectively, for each fluoxetine enantiomer; the correlation coefficient was >0.990. Method ruggedness was demonstrated by the reproducible performance of the assay during a three-day validation period. PMID- 11857716 TI - Automated system for simultaneous analysis of delta(13)C, delta(18)O and CO(2) concentrations in small air samples. AB - In this paper we present an automated system for simultaneous measurement of CO(2) concentration, delta(13)C and delta(18)O from small (<1 mL) air samples in a short period of time (approximately 1 hour). This system combines continuous flow isotope ratio mass spectrometry (CF-IRMS) and gas chromatography (GC) with an inlet system similar to conventional dual-inlet methods permitting several measurement cycles of standard and sample air. Analogous to the dual-inlet method, the precision of this system increases with the number of replicate cycles measured. The standard error of the mean for a measurement with this system is 0.7 ppm for the CO(2) concentration and 0.05 per thousand for the delta(13)C and delta(18)O with four replicate cycles and 0.4 ppm and 0.03 per thousand respectively with nine replicate cycles. The mean offset of our measurements from NOAA/CMDL analyzed air samples was 0.08 ppm for the CO(2) concentration, 0.01 per thousand for delta(13)C and 0.00 per thousand for delta(18)O. A specific list of the parts and operation of the system is detailed as well as some of the applications for micrometeorological and ecophysiological applications. PMID- 11857717 TI - Characterisation and determination of indole alkaloids in frog-skin secretions by electrospray ionisation ion trap mass spectrometry. AB - The characterisation of selected indole alkaloids in a quadrupole ion trap mass spectrometer is presented. Fragmentation profiles for tryptamine, 5 hydroxytryptamine (5-HT), N'-methyl 5-hydroxytryptamine (N'-methyl 5-HT), N',N' dimethyl 5-hydroxytryptamine (bufotenine), N',N',N'-trimethyl 5-hydroxytryptamine (5-HTQ), and N',N'-dimethyl 5-methoxytryptamine (5-MeODMT) are presented with proposed structures given for each product ion observed. Such MS(n) experiments can be used to differentiate the isobaric molecular ions of the compounds 5-HTQ (M(+)) and 5-MeODMT (MH(+)). The quantitative determination of certain indole alkaloids in the skin secretions of the Australian Golden Bell frog, Litoria aurea, by LC/ESI-ion trap MS is also presented. The concentrations of 5-HT, N' methyl 5-HT and 5-HTQ were found to be 2.68, 0.26 and 0.54 microg per mg of skin secretion, respectively. PMID- 11857719 TI - Analysis of the saliva from patients with oral cancer by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. AB - Using matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS), this study analyzed the saliva obtained from patients with oral cancer and compared these mass spectra with those obtained from healthy controls. Saliva without pre-treatment was mixed directly with a sinapinic acid matrix. Alpha amylase (57 kDa) dominated the high mass range in the MALDI mass spectra of the saliva from healthy subjects, but the peak was suppressed for patients with oral cancer and was replaced by a peak at m/z 66 k in the spectra of patients' samples (15 out of 20). Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PAGE) with in-gel tryptic digestion combined with matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) was employed to characterize this 66-kDa protein, which was thus shown to be albumin. However, based on SDS PAGE results, concentrations of both alpha-amylase and albumin in patients' saliva were significantly higher than those in healthy subjects. This discrepancy was shown to be due to MALDI suppression effects due to the albumin. MALDI-MS thus has potential as a possible rapid diagnostic screening tool for oral cancer. PMID- 11857718 TI - Cluster ions derived from sodium and potassium tetrafluoroborate and their collision induced dissociation in an ion trap mass spectrometer. AB - Electrospray ionization was used to produce distributions of gas-phase cluster ions from solutions of sodium and potassium tetrafluoroborate. The majority of the cluster species followed the trend (MBF(4))(n)M(+), where M=Na and K. The values of n, for both salts, ranged from 1-15. Collision induced dissociation (MS/MS and MS(n)) in an ion trap mass spectrometer was used to determine the dissociation pathways for the cluster ions. The (NaBF(4))(n)Na(+) cluster ions fragmented via two pathways: (a) the loss of one or multiple neutral BF(3) molecules and (b) the loss of one or more NaBF(4) units. Of the two, the product ions corresponding to the loss of BF(3) units were more prominent. Unlike the Na salt, the (KBF(4))(n)K(+) cluster ions decomposed primarily by the loss of one or multiple KBF(4) units. Similar differences in dissociation behavior were observed when the heated transfer capillary, normally used to desolvate ions, was used to investigate cluster ion stability to thermal degradation and dissociation. The dissociation profiles (decrease in ion abundance with increasing activation amplitude) for several (NaF)(n)Na(+) and (KF)(n)K(+) cluster ions were measured and compared to probe the influence of the relative stability of the alkali fluorides (NaF and KF) on the dissociation behavior exhibited by the tetrafluoroborate cluster distributions. We found that the (NaF)(n)Na(+) cluster ions required higher activation amplitudes to induce fragmentation than the corresponding (KF)(n)K(+) species, indicative of stronger ionic bonding and higher gas-phase stability for the former. This in turn indicates that the reaction pathway involving only the loss of one or multiple units of BF(3), favored for the (NaBF(4))(n)Na(+) cluster series, but not for the analogous (KBF(4))(n)K(+) series, may be due to the high gas-phase stability of NaF, and relatively lower stability of KF, towards dissociation. PMID- 11857720 TI - Urinary 19-norandrosterone purification by immunoaffinity chromatography: application to gas chromatography/combustion/isotope ratio mass spectrometric analysis. AB - The detection of exogenous 19-norandrosterone (19-NA) in urines was investigated by using gas chromatography/combustion/isotope ratio mass spectrometry (GC/C/IRMS). 19-NA is, for the first time to our knowledge, isolated from urinary matrix by specific immunoaffinity chromatography (IAC) before analysis. The sample preparation consisted of a preliminary purification of urine by solid phase extraction after hydrolysis by beta-glucuronidase. Unconjugated 19-NA was thus isolated by IAC and directly analysed by GC/C/IRMS. Optimisation of IAC purification was achieved and the reliability of the technique for anti-doping control is discussed. PMID- 11857722 TI - Loss of isocyanic acid from the internal oxadiazole ring of protonated molecules of some 2,5-diaryl-1,3,4-oxadiazoles. AB - The fragmentation pattern of some protonated 2,5-diaryl-1,3,4-oxadiazoles is discussed. An unusual decomposition consisting of elimination of the isocyanic acid molecule from the internal oxadiazole ring was found. This fragmentation pathway was deduced on the basis of B/E linked scan mass spectra of metastable ions with liquid secondary ion mass spectrometry as the ionization method and also of low-energy CID mass spectra where electrospray was used as the ionization technique. High resolution measurements were also performed. PMID- 11857721 TI - Combined quantum chemical and RRKM modeling of the main fragmentation pathways of protonated GGG. II. Formation of b(2), y(1), and y(2) ions. AB - Quantum chemical and RRKM calculations were performed on protonated GGG in order to determine the atomic details of the main fragmentation pathways leading to formation of b(2),y(1), and y(2) ions. Formation of y(1) ions on the "diketopiperazine" pathway is initiated from relatively high-energy C-terminal amide nitrogen protonated species for which the N-terminal amide bond is in the cis isomerization state. The reaction goes through a transition structure which is only slightly less favored than the reactive configuration itself. RRKM calculations indicate that this reaction is extremely fast as soon as the fragmenting species have more internal energy than the reaction threshold. The calculated energetics suggests that y(1) ions are formed on the "diketopiperazine" pathway with a non-negligible (6-10 kcal/mol) reverse activation barrier. Investigation of species occurring during the formation of b(2) ions having an oxazolone structure indicates that y(1) ions can be formed also from intermediates previously thought to result in only b(2) ions. As the first step of the "b(x)-y(z)" pathway proposed here the extra proton must reach the nitrogen of the C-terminal amide bond. Attack of the N-terminal amide oxygen on the carbon center of the C-terminal amide bond results in formation of the oxazolone ring while the detaching G leaves the precursor ion. Under low-energy collision conditions the complex of protonated 2-aminomethyl-5-oxazolone and G can rearrange to form a proton-bonded dimer of these species. In such circumstances the extra proton is shared by the two monomers and dissociation of the dimer will be determined by the thermochemistry involved. Based on the "b(x) y(z)" pathway one can easily explain the linear relationship between the logarithm of the y(1)/b(2) ion abundance ratio and the proton affinity of the C terminal amino acid substituent for the series of H-Gly-Gly-Xxx-OH tripeptides where Xxx was varied (Morgan DG, Bursey MM. Org. Mass. Spectrom. 1994; 29: 354). The calculated energetics indicates that both y(1) and b(2) ions are formed with no reverse activation barrier on the "b(x)-y(z)" pathway. PMID- 11857723 TI - Determination of naphthodianthrones and phloroglucinols from Hypericum perforatum extracts by liquid chromatography/tandem mass spectrometry. AB - Hypericum perforatum L. (St. John's Wort) has long been known as a medicinal plant, and has been used for the treatment of depression and neuralgic disorders. Its main active constituents are believed to be a naphthodianthrone, hypericin, and a phloroglucinol, hyperforin. A sensitive high performance liquid chromatography (HPLC)/electrospray tandem mass spectrometric method for fast simultaneous determination of six major naphthodianthrones and phloroglucinols of Hypericum perforatum extract has been developed. The method, based on multiple dissociation reaction monitoring (MRM), allows the analysis of hypericin, protohypericin, pseudohypericin, protopseudo-hypericin, hyperforin and adhyperforin from the extract in less than 5 min. Good linearity over the range 0.1-1000 ng/mL for hyperforin and 2-500 ng/mL for hypericin was observed. Intra assay accuracy and precision varied from 2 to 19% within these ranges. Lower levels of quantitation for hyperforin were 0.5 ng/mL and 2 ng/mL for hypericin. PMID- 11857724 TI - Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and high-performance liquid chromatography study of quantitative and qualitative variation in tarantula spider venoms. AB - Animal venoms are important sources of novel pharmacological tools, useful in biochemical characterization of their receptors. Venom quality control, batch-to batch homogeneity and high reproducibility of venom fractionation and toxin purification are crucial issues for biochemical and pharmacological studies. To address these issues, a study of the variability of tarantula spider venom samples was undertaken. Venom profiles of samples collected from individuals of different age and sex, and from sibling spiders of the same species, were generated by high-performance liquid chromatography (HPLC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) and analyzed to assess venom variability and method accuracy. Sex-linked venom variation was studied on eight species. Clear qualitative differences were observed for six out of eight species, as well as quantitative differences. Age related variation studied in Poecilotheria rufilata showed essentially age related quantitative differences between adults of both sexes and immature juveniles. The venoms of nine siblings and three wild-collected Pterinochilus murinus were studied for individual variation, showing only very minor quantitative differences. On the same samples, the quality of MALDI-TOFMS venom fingerprinting was demonstrated to be highly reproducible. Our results show that tarantula venom peptide fingerprinting is a highly reliable identification method, that pooled batches of venom from several animals can be used for venom purification, that venom composition does not appear to be qualitatively related to ontogenesis in the spiders studied, and that qualitative sex-linked variation occurs across most species and may be important in activity studies. PMID- 11857725 TI - Fragmentation studies on monensin A and B by accurate-mass electrospray tandem mass spectrometry. AB - Monensin A and B were studied by electrospray ionisation tandem mass spectrometry (ESI-MS/MS) and the fragment ions were confirmed by accurate-mass measurements. Analyses were performed on both a quadrupole time-of-flight (QTOF) and a Fourier transform ion cyclotron resonance (FTICR) mass spectrometer. The analysis revealed that fragment ions were produced by Grob-Wharton fragmentations and pericyclic rearrangements in addition to various simple neutral losses. A study of the protonated and sodiated sodium salt revealed different fragmentation pathways for these species, thus complementary structural information could be gained. A complete fragmentation pathway of monensin A and B protonated sodium salt [(M-H+Na)+H])+) and sodiated sodium salt [(M-H+Na)+Na](+) is proposed. MS(3) analysis confirmed the separate fragmentation pathways. PMID- 11857726 TI - Generic method for on-line extraction of drug substances in the presence of biological matrices using turbulent flow chromatography. AB - The use of liquid chromatography/mass spectrometry (LC/MS) to quantify drugs in biological matrices has been well established over the last decade. Extremely fast LC/MS methods are commonplace in the pharmaceutical industry for high throughput Absorption, Distribution, Metabolism and Excretion (ADME) screening. However, to truly take full advantage of high-throughput ADME screening, a generic method is needed that eliminates the need to develop a new method for each new compound being screened. New developments in the stationary phase of turbulent flow columns has allowed us to develop an on-line biological sample cleanup method that is suitable for over 99% of the compounds in the Cephalon database. PMID- 11857728 TI - Derivatization for liquid chromatography/electrospray mass spectrometry: synthesis of tris(trimethoxyphenyl)phosphonium compounds and their derivatives of amine and carboxylic acids. AB - A simple method for the derivatization of primary amines and carboxylic acids with tris(trimethoxyphenyl)phosphonium (TMPP) reagents to enhance their detection by electrospray mass spectrometry (ESI-MS) has been developed. The synthesis of novel TMPP reagents and their stable isotopically labelled analogues is described. Through the use of stable isotopically labelled TMPP "tags", incorporation of a doublet (1:1, (1)H/(2)H or (12)C/(13)C) into the target molecule can be achieved, enabling the use of isotopic target analysis to detect compounds of unknown molecular weight but with a characteristic isotope pattern and accurate mass difference. PMID- 11857727 TI - In situ analysis of solvents on breath and blood: a selected ion flow tube mass spectrometric study. AB - We report measurements of residual vapour levels of xylenes and trimethylbenzenes, present following a floor re-surfacing procedure, using the technique of selected ion flow tube mass spectrometry (SIFT-MS). A subject exposed to controlled amounts of xylene and mesitylene was monitored by direct breath exhalation over a 4-hour period after exposure to the volatile organic compounds (VOCs) had stopped. The headspace gases above 5 mL blood samples taken over this period were also monitored. The decays of the solvent levels with time were fitted to a two-compartment model with residence times for xylene and mesitylene of 0.37 h and 0.38 h, respectively (compartment one) and 2.5 h and 2.8 h, respectively (compartment two). PMID- 11857729 TI - Short pulse laser mass spectrometry of nitrotoluenes: ionization and fragmentation behavior. AB - The mass spectra of all isomers of mononitrotoluene, four isomers of dinitrotoluene and of 2,4,6,-trinitrotoluene, obtained by multiphoton ionization utilizing ultrashort laser pulses with center wavelengths of either 206 nm or 412 nm, are presented and discussed. Under these ionization conditions all nitrotoluenes exhibit a high degree of fragmentation which increases with the degree of substitution. For the compounds having a nitro group in position 2 and/or 6 a pronounced ortho effect leading to the loss of OH is observed. The fragmentation patterns in the lower mass range are typical for alkylated aromatic substances. While no fundamental differences between the mass spectra obtained with the two wavelengths were observed, the visible light in all cases resulted in a broader variety of fragments and additional signals in the higher mass range. The latter can be used for isomer identification. PMID- 11857730 TI - Structural isomer differentiation of 1,6-diaza-3,8-dioxabicyclo[4.4.1]undecane and N,N'-methylenebis(oxazolidine) by mass spectrometry. AB - The title compound types are structural isomers. The absence of literature studies dealing with the mass spectrometric gas-phase behavior of such compounds prompted a search for the best conditions for differentiation between the two prototype examples (R,R)-5,10-diphenyl-1,6-diaza-3,8-dioxabi-cyclo[4.4.1]undecane and N,N'-methylenebis[(R)-4-phenyloxazolidine]. Attempts to differentiate between the isomers by NMR were inconclusive and X-ray crystallography had to be used. The best mass spectrometric results were obtained using gas-phase ionization techniques, particularly chemical ionization, whereby protonated molecules were observed. The fragmentation pathways were elucidated from MS/MS studies and from experiments performed on hexadeuterated samples of each compound. PMID- 11857731 TI - Atmospheric pressure chemical ionisation liquid chromatography/mass spectrometry of bacteriochlorophylls from Chlorobiaceae: characteristic fragmentations. AB - Atmospheric pressure chemical ionisation liquid chromatography/mass spectrometry/mass spectrometry (APCI-LC/MS/MS) has been applied to the study of bacteriochlorophylls c, d, and e of phototrophic prokaryotes. Cultures of Chlorobiaceae containing bacteriochlorophyll c, d or e were examined using a high resolution high-performance liquid chromatography (HPLC) method and APCI-LC/MS/MS employing post-column addition of formic acid. The results reveal complex distributions of bacteriochlorophyll homologues, with some closely eluting species giving isobaric protonated molecules. On-line LC/MS/MS studies reveal characteristic fragment ions for bacteriochlorophylls c, d, and e. Fragmentations involving loss of the extended alkyl substituents that are unique to bacteriochlorophylls c, d and e and their derivatives have been rationalised by studying the phaeophorbides and the results applied to the direct study of the bacteriochlorophylls. PMID- 11857732 TI - New algorithms for processing and peak detection in liquid chromatography/mass spectrometry data. AB - Two new algorithms for automated processing of liquid chromatography/mass spectrometry (LC/MS) data are presented. These algorithms were developed from an analysis of the noise and artifact distribution in such data. The noise distribution was analyzed by preparing histograms of the signal intensity in LC/MS data. These histograms are well fit by a sum of two normal distributions in the log scale. One new algorithm, median filtering, provides increased performance compared to averaging adjacent scans in removing noise that is not normally distributed in the linear scale. Another new algorithm, vectorized peak detection, provides increased robustness with respect to variation in the noise and artifact distribution compared to methods based on determining an intensity threshold for the entire dataset. Vectorized peak detection also permits the incorporation of existing algorithms for peak detection in ion chromatograms and/or mass spectra. The application of these methods to LC/MS spectra of complex biological samples is described. PMID- 11857733 TI - Measurements by gas chromatography/pyrolysis/mass spectrometry: fundamental conditions in (2)H/(1)H isotope ratio analysis. AB - Gas chromatography/pyrolysis/isotope ratio mass spectrometry (GC/P/IRMS) is a relatively new method for on-line determination of (2)H/(1)H isotope ratios. The influence of different parameters on the (2)H/(1)H isotope ratios obtained in GC/P/IRMS has been thoroughly studied using several flavor compounds, such as 5 nonanone, linalool, menthol, linalyl acetate and 4-decanolide. The requirement of "conditioning" the pyrolysis reactor to obtain reliable delta(2)H(V-SMOW) values is discussed. Furthermore, the influence of the carrier gas flow of the gas chromatograph on the completeness of pyrolysis and subsequently on the delta(2)H(V-SMOW) values is investigated in detail. The linear range of the compounds investigated is determined. The results show that calibrating the GC/P/IRMS system with secondary standard substances is absolutely necessary in order to obtain reliable delta(2)H(V-SMOW) values. In view of interlaboratory comparability, validation procedures are recommended. PMID- 11857734 TI - Atmospheric pressure chemical ionisation normal phase liquid chromatography mass spectrometry and tandem mass spectrometry of chlorophyll a allomers. AB - Four oxidation products formed during the allomerisation reaction of chlorophyll a in methanol have been separated and identified using normal phase high performance liquid chromatography (NP-HPLC) coupled with atmospheric pressure chemical ionisation (APCI) tandem mass spectrometry (MS/MS) and photodiode array detection. Fragmentation pathways of the main stereoisomers of the four major products, Mg-purpurin-7 dimethyl phytyl ester, 15(1)-MeO-lact-chl a, 13(2)-MeO chl a and 13(2)-HO-chl a, have been assigned using APCI-MS(n) aided by examination of deuteriated counterparts of the purpurin and the lactone. Diagnostic fragmentations have been determined, including those for Mg-purpurin-7 dimethyl phytyl ester and 15(1)-MeO-lact-chl a, which display very similar full MS and MS/MS spectra. The use of NP-HPLC offers a rapid and efficient method which, combined with MS/MS, reduces reliance on precursor ion mass for assignment of structure in the allomerisation reaction where major and minor components are often isobaric. PMID- 11857735 TI - Differential occurrence of mutations causative of eye diseases in the Chinese population. AB - Ethnic differences and geographic variations affect the frequencies and nature of human mutations. In the literature, descriptions of causative mutations of eye diseases in the Chinese population are few. In this paper we attempt to reveal molecular information on genetic eye diseases involving Chinese patients from published and unpublished works by us and other groups. Our studies on candidate genes of eye diseases in the Chinese population in Hong Kong include MYOC and TISR for primary open angle glaucoma, RHO and RP1 for retinitis pigmentosa, ABCA4 and APOE for age-related macular degeneration, RB1 for retinoblastoma, APC for familial adenomatous polyposis with congenital hypertrophy of retinal pigment epithelium, BIGH3/TGFBI for corneal dystrophies, PAX6 for aniridia and Reiger syndrome, CRYAA and CRYBB2 for cataracts, and mtDNA for Leber hereditary optic neuropathy. We have revealed novel mutations in most of these genes, and in RHO, RP1, RB1, BIGH3, and PAX6 we have reported mutations that contribute to better understanding of the functions and properties of the respective gene products. We showed absence of MYOC does not necessarily cause glaucoma. No disease causative mutations have been identified in MYOC or ABCA4. There are similarities in the patterns of sequence alterations and phenotype-genotype associations in comparison with other ethnic groups, while the MYOC, RB1, APC, and PAX6 genes have more Chinese-specific sequence alterations. Establishment of a mutation database specific for the Chinese is essential for identification of genetic markers with diagnostic, prognostic, or pharmacological values. PMID- 11857736 TI - Mutations in the RUNX2 gene in patients with cleidocranial dysplasia. AB - Cleidocranial dysplasia (CCD) is a autosomal dominant disorder characterized by skeletal anomalies such as patent fontanels, late closure of cranial sutures with Wormian bones, late erupting secondary dentition, rudimentary clavicles, and short stature. The locus for this disease was mapped to chromosome 6p21. RUNX2 is a member of the runt family of transcription factors and its expression is restricted to developing osteoblasts and a subset of chondrocytes. Mutations in the RUNX2 gene have been shown to cause CCD. Chromosomal translocations, deletions, insertions, nonsense and splice-site mutations, as well as missense mutations of the RUNX2 gene have been described in CCD patients. Although there is a wide spectrum in phenotypic variability ranging from primary dental anomalies to all CCD features plus osteoporosis, no clear phenotype-genotype correlation has been established. However analysis of the three-dimensional structure of the DNA binding runt domain of the RUNX proteins and its interaction with DNA, as well as the cofactor CBFB, start to provide an insight into how missense mutations affect RUNX2 function. PMID- 11857737 TI - G6PDdb, an integrated database of glucose-6-phosphate dehydrogenase (G6PD) mutations. AB - G6PDdb (http://www.rubic.rdg.ac.uk/g6pd/ or http://www.bioinf.org.uk/g6pd/) is a newly created web-accessible locus-specific mutation database for the human Glucose-6-phosphate dehydrogenase (G6PD) gene. The relational database integrates up-to-date mutational and structural data from various databanks (GenBank, Protein Data Bank, etc.) with biochemically characterized variants and their associated phenotypes obtained from published literature and the Favism website. An automated analysis of the mutations likely to have a significant impact on the structure of the protein has been performed using a recently developed procedure. The database may be queried online and the full results of the analysis of the structural impact of mutations are available. The web page provides a form for submitting additional mutation data and is linked to resources such as the Favism website, OMIM, HGMD, HGVBASE, and the PDB. This database provides insights into the molecular aspects and clinical significance of G6PD deficiency for researchers and clinicians and the web page functions as a knowledge base relevant to the understanding of G6PD deficiency and its management. PMID- 11857738 TI - HbVar: A relational database of human hemoglobin variants and thalassemia mutations at the globin gene server. AB - We have constructed a relational database of hemoglobin variants and thalassemia mutations, called HbVar, which can be accessed on the web at http://globin.cse.psu.edu. Extensive information is recorded for each variant and mutation, including a description of the variant and associated pathology, hematology, electrophoretic mobility, methods of isolation, stability information, ethnic occurrence, structure studies, functional studies, and references. The initial information was derived from books by Dr. Titus Huisman and colleagues [Huisman et al., 1996, 1997, 1998]. The current database is updated regularly with the addition of new data and corrections to previous data. Queries can be formulated based on fields in the database. Tables of common categories of variants, such as all those involving the alpha1-globin gene (HBA1) or all those that result in high oxygen affinity, are maintained by automated queries on the database. Users can formulate more precise queries, such as identifying "all beta-globin variants associated with instability and found in Scottish populations." This new database should be useful for clinical diagnosis as well as in fundamental studies of hemoglobin biochemistry, globin gene regulation, and human sequence variation at these loci. PMID- 11857739 TI - The A8296G mtDNA mutation associated with several mitochondrial diseases does not cause mitochondrial dysfunction in cybrid cell lines. AB - Transmitochondrial cybrid cell lines homoplasmic for the A8296G mtDNA transition, a mutation associated with several mitochondrial diseases, have a normal oxidative phosphorylation function, as shown by oxygen consumption, lactate production, respiratory enzyme activities, and growth using galactose as the only source of energy. The synthesis of mitochondrial proteins is also similar in mutant and wild-type cybrids. Our results suggest that the A8296G mutation is a polymorphism and reinforce the necessity of performing functional studies to assess the pathogenicity of mtDNA mutations. PMID- 11857740 TI - Mutation detection in the duplicated region of the polycystic kidney disease 1 (PKD1) gene in PKD1-linked Australian families. AB - Screening for disease-causing mutations in the duplicated region of the PKD1 gene was performed in 17 unrelated Australian individuals with PKD1-linked autosomal dominant polycystic kidney disease. Exons 2-21 and 23-34 were assayed using PKD1 specific PCR amplification and direct sequencing. We have identified 12 novel probably pathogenic DNA variants, including five truncating mutations (Q563X, c.5105delAT, c.5159delG, S2269X, c.9847delC), two in-frame deletions (c.7472del3, c.9292del39), and two splice-site mutations (IVS14+1G>C, IVS16+1G>T). Three of the mutations (G381C, Y2185D, G2785D) were predicted to lead to the replacement of conserved amino acid residues, with ensuing changes in protein conformation. Defects in the duplicated region of PKD1 thus account for 63% of our patients. Together with the previously detected mutations (Q4041X, R4227P) in the 3 region of the gene, the study has achieved an overall mutation detection rate of 74%. In addition, we have detected 31 variants (nine novel and 22 previously published) that did not segregate with the disease and were considered to be neutral polymorphisms. Three of the nine novel polymorphisms were missense mutations with a predicted effect on protein conformation, emphasizing the problems of interpretation in PKD1 mutation screening. PMID- 11857741 TI - Identification of 33 polymorphisms in the adipocyte-derived leucine aminopeptidase (ALAP) gene and possible association with hypertension. AB - Adipocyte-derived leucine aminopeptidase (ALAP) inactivates angiotensin II and/or generates bradykinin in the kidney, suggesting a possible role for ALAP in the regulation of blood pressure. We considered the hypothesis that genomic variants of the ALAP gene are associated with hypertension or individual variations in blood pressure. We screened for mutations in the ALAP gene in 48 unrelated Japanese individuals and identified 33 polymorphisms including 15 novel polymorphisms. We then performed a two-stage analysis. In the first stage, the eight missense polymorphisms were evaluated for associations with blood pressure in 96 apparently healthy individuals. In the second stage, only the most promising polymorphisms were evaluated for association with essential hypertension in 143 hypertensive and 348 normotensive subjects. Among the eight missense polymorphisms, the Ile276Met and Lys528Arg polymorphisms showed significant association with blood pressure. Subsequent analysis confirmed association between the Lys528Arg polymorphism and essential hypertension. The estimated odds ratio for essential hypertension was 2.3 for presence of the Arg allele at codon 528, in comparison with presence of the Lys/Lys genotype (P = 0.004). These findings support involvement of ALAP in the regulation of blood pressure. PMID- 11857743 TI - Identification and in vitro expression of novel CDH23 mutations of patients with Usher syndrome type 1D. AB - Usher syndrome (USH) is a group of autosomal recessive sensory disorders characterized by progressive retinitis pigmentosa (RP) and sensorineural hearing impairment. Usher syndrome type 1 (USH1), with additional vestibular dysfunction, represents the most severe form and shows extensive allelic and non-allelic heterogeneity. At least six USH1 loci exist (USH1A-F), and four of the underlying genes have been identified. Recently, a novel gene, cadherin 23 (CDH23), was shown to be mutated in USH1D. We performed mutation screening by single strand conformation polymorphism (SSCP) analysis and direct sequencing on 33 USH1 patients previously excluded for USH1B and USH1C. On eight disease alleles of four patients, four different mutations were identified, three of them novel (c.6933delT, c.5712G-->A, and IVS45-9G-->A). Exon trapping experiments were performed with two mutations. In the case of a c.5712G-->A transition of the last base of exon 42, that is an apparently synonymous mutation, skipping of exon 42 was observed. By the mutation IVS45-9G-->A, a novel splice acceptor site was created and the insertion of 7 intronic bp was observed. Two mutations, IVS45-9G- >A and the previously described IVS51+5G-->A, were each found in more than one patient. Haplotype analysis by SNPs within CDH23 suggests common ancestors for each of the mutations. Among the total of 52 USH1 cases studied by us, CDH23 mutations account for about 10% of all disease alleles. Our results further suggest that in patients with a typical USH1D phenotype, a significant portion of CDH23 mutations leads to premature termination of translation or loss of numerous amino acid residues, with a high frequency of changes causing aberrant splicing of CDH23 mRNA. PMID- 11857742 TI - Structure, genomic DNA typing, and kinetic characterization of the D allozyme of placental alkaline phosphatase (PLAP/ALPP). AB - The D allozyme of placental alkaline phosphatase (PLAP) displays enzymatic properties at variance with those of the common PLAP allozymes. We have deduced the amino acid sequence of the PLAP D allele by PCR cloning of its gene, ALPP. Two coding substitutions were found in comparison with the cDNA of the common PLAP F allele, i.e., 692C>G and 1352A>G, which translate into a P209R and E429G substitution. A single nucleotide primer extension (SNuPE) assay was developed using PCR primers that enable the amplification of a 1.9 kb PLAP fragment. Extension primers were then used on this PCR fragment to detect the 692C>G and 1352A>G substitution. The SNuPE assay on these two nucleotide substitutions enabled us to distinguish the PLAP F and D alleles from the PLAP S/I alleles. Functional studies on the D allozyme were made possible by constructing and expressing a PLAP D cDNA, i.e., [Arg209, Gly429]PLAP, into wild-type Chinese hamster ovary cells. We determined the k(cat) and K(m), of the PLAP S, F, and D allozymes using the non-physiological substrate p-nitrophenylphosphate at an optimal pH (9.8) as well as two physiological substrates, i.e., pyridoxal-5 phosphate and inorganic pyrophosphate at physiological pH (7.5). We found that the biochemical properties of the D allozyme of PLAP are significantly different from those of the common PLAP allozymes. These biochemical findings suggest that a suboptimal enzymatic function by the PLAP D allozyme may be the basis for the apparent negative selective pressure of the PLAP D allele. The development of the SNuPE assay will enable us to test the hypothesis that the PLAP D allele is subjected to intrauterine selection by examining genomic DNA from statistically informative population samples. PMID- 11857744 TI - The identification and classification of 41 novel mutations in the factor VIII gene (F8C). AB - Hemophilia A is a bleeding disorder caused by a quantitative or qualitative deficiency in the coagulation factor VIII. Causative mutations are heterogeneous in nature and are distributed throughout the FVIII gene. With the exception of mutations that result in prematurely truncated protein, it has proved difficult to correlate mutation type/amino acid substitution with severity of disease. We have identified 81 mutations in 96 unrelated patients, all of whom have typed negative for the common IVS-22 inversion mutation. Forty-one of these mutations are not recorded on F8C gene mutation databases. We have analyzed these 41 mutations with regard to location, whether or not each is a cross-species conserved region, and type of substitution and correlated this information with the clinical severity of the disease. Our findings support the view that the phenotypic result of a mutation in the FVIII gene correlates more with the position of the amino acid change within the 3D structure of the protein than with the actual nature of the alteration. PMID- 11857746 TI - Reliability of DHPLC in mutational screening of beta-globin (HBB) alleles. AB - The inherited disorders of hemoglobin represent the most common Mendelian disease worldwide, with a higher prevalence among Mediterraneans, Asians, Africans, and Indians. Altered beta-globin sequences, causing either hemoglobinopathies or beta thalassemia syndromes, are due to more than 200 different mutations in the beta globin gene. Prevention programs based on postnatal and prenatal molecular diagnosis of heterozygous carriers and/or patients require the use of reliable mutation scanning methods in at-risk populations. We have developed a rapid and highly specific mutation screening test based on the denaturing high-performance liquid chromatography (DHPLC) system. The sensitivity and specificity of the method were tested on the full genomic region of the beta-globin gene in 30 normal Italian subjects and 40 heterozygous carriers in which 25 different beta globin mutations had been previously characterized by multiplex-ARMS technique. The results showed DHPLC to be 100% sensitive and specific. All the 25 sequence alterations and two previously undetected polymorphisms were precisely identified with neither false positive nor false negative results. In addition, 12 compound heterozygous and four homozygous patients were successfully subjected to DHPLC. Overall, the method was able to rapidly identify the most common beta-globin mutations, accounting for more than 97% of beta-globin alleles in the Italian population. Compared to classical approaches of mutation screening, this method allows a rapid, highly sensitive, cost-effective, and semi-automated simultaneous mutational scanning of a large number of samples. PMID- 11857745 TI - A modified multiplex PCR assay for detection of large deletions in MSH2 and MLH1. AB - A method for detection of large genomic deletions in the MSH2 and MLH1 genes based on multiplex PCR and quantitative evaluation of PCR products is presented. All 35 exons of MSH2 and MLH1 were screened simultaneously in seven PCR reactions, each of them including primers for both genes. The method is reliable for uncovering large genomic deletions in patients suspected of HNPCC. With this method, six novel deletions were identified, two in MSH2: EX1_10del and EX1_16del (representing deletion of the entire MSH2 gene); and four in MLH1: EX1_10del in two unrelated patients, EX3_5del, and EX4del. The deletions were detected in 18 unrelated patients in whom no germline mutation had been identified by SSCP and DHPLC. These results indicate that our modified multiplex PCR assay is suited for the detection of large deletions both in the MSH2 and MLH1 gene and therefore represents an additional valuable tool for mutation screening in HNPCC families. PMID- 11857748 TI - Low frequency of recurrent BRCA1 and BRCA2 mutations in Spain. AB - BRCA1 and BRCA2 mutations underlie a substantial proportion of all hereditary breast cancer. The mutational spectrum in these genes is very broad, with hundreds of different BRCA mutations reported worldwide. However, high frequency founder mutations make up a substantial fraction of all mutations in some ethnic groups. We directly sequenced BRCA1 and BRCA2 in 35 Spanish breast/ovarian cancer families and found 13 mutations of which 3 had been reported previously in Spain. The ten novel mutations are: IVS5+1 G>A, 1491delA, Leu1086Ter, and Gln895Ter in BRCA1; Glu49Ter, 5373delGTAT, 5947delCTCT, 6672delTA, 8281insA, and Pro3039Leu (which also involves a splice site) in BRCA2. Our data, in combination with previous reports, indicate that 14 mutations have been seen recurrently in Spanish families. Analyzing these 14 mutations in 42 previously untested breast/ovarian cancer families revealed only two families testing positive, one for BRCA1 185delAG and one for BRCA2 9254delATCAT. While several mutations have been found recurrently in Spain, none appear to be high frequency founder mutations based on studies of breast and ovarian cancer families. PMID- 11857747 TI - A new strategy for mannose-binding lectin gene haplotyping. AB - The mannose-binding lectin 2 (MBL2) gene is polymorphic and codes for a protein with an important role in the innate immune response, whose variants have been associated with a great number of diseases. Point variations have been described in the 5' regulatory region at positions -550 (MBL2*H or *L) and -221 (*X or *Y), in the 5' untranslated sequence at position +4 (*P or *Q), and in the coding sequence of exon 1 at codons 52, 54, and 57 (MBL2*A or D, A or B, and A or C, respectively). These can be in cis or in trans configuration. The different haplotypes influence the immunological phenotype of the individual, which makes MBL2 haplotyping very important. Previously described MBL2-typing methods do not present adequate haplotype resolution or are too complex and costly. We have developed a new MBL2-typing strategy that is economical and renders rapid and reliable results without ambiguities. We typed 202 individuals of European, 32 of African, and 16 of Oriental descent. Only five to six reactions from 10 possible PCR-SSPs (sequence-specific polymerase chain reactions) were sufficient to genotype one individual unambiguously. The reactions were specific for amplification of the variants located upstream of the coding sequence. The results were associated to the results of hybridizations of the amplified products with eight sequence-specific oligonucleotide probes (SSOP). The strategy led to identification of eight alleles: MBL2*HYPA, HYPD, LYPA, LYPB, LYPD, LYQA, LYQC, and LXPA. Their frequencies in each of the groups were similar to those of other populations studied to date, with MBL2*LYPD (g.[-550G>C; -221C>G; 4T>C; 223C>T; 230A>G; 239A>G]) being novel. All samples were found to be in Hardy Weinberg equilibrium. PMID- 11857749 TI - Recurrent BRCA1 and BRCA2 germline mutations in ovarian cancer: a founder mutation of BRCA1 identified in the Chinese population. AB - Previous mutational analysis for BRCA gene mutations in sporadic ovarian cancer occurring in Chinese patients in Hong Kong identified six germline BRCA1 mutations and one germline BRCA2 mutation, six of which were novel (Khoo et al., 2000). Knowledge of BRCA gene mutations in the Chinese population is relatively scant. In this study, we focussed on whether any of these mutations could be recurrent in our Chinese population, making use of archival paraffin embedded tissue. A consecutive series of 214 ovarian cancer cases, half of Southern Chinese origin from Hong Kong whilst the other half of Northern Chinese origin from Beijing were used for the study. We identified one further novel mutation, 1081delG, in BRCA1. This was found to occur in two unrelated individuals with shared haplotype as revealed by allelotype analysis, thus demonstrating founder effect. Two other recurrent mutations were also identified, the 2371-2372delTG mutation in BRCA1 and the 3337C>T mutation in BRCA2 recurring in two and three unrelated individuals respectively, giving an overall prevalence 4.7% of recurrent BRCA mutations in ovarian cancer in the Southern Chinese population. Most importantly, all our recurrent mutation carriers were identified from Southern Chinese patients from Hong Kong whilst such mutations were absent in samples from the Northern Chinese. Our findings indicate possible heterogeneity in the BRCA genotype between Northern and Southern Chinese. The identification of a founder mutation and two recurrent mutations moreover, has important implications towards screening strategies for breast and ovarian cancer among Chinese of southern ancestral origin who are now dispersed throughout the world. PMID- 11857750 TI - Identification of a novel NOG gene mutation (P35S) in an Italian family with symphalangism. AB - Symphalangism (SYM or SYM1) is an autosomal dominant disorder characterized by multiple joint fusions. The disease is caused by mutations of the NOG gene, that maps to chromosome 17q22. So far, only six independent NOG mutations have been identified. We have analysed an Italian family in which father and son had bilateral symphalangism and detected a novel NOG mutation (P35S), originated in the father from a c.914C>T transition. A different mutation in the same codon (P35R) has been previously described. Comparison between different noggin gene hortologs shows that codon 35 is conserved. Therefore, this codon should play an important role in NOG gene function. This is the first mutation described for NOG after the initial report of NOG mutations being causative of SYM. PMID- 11857751 TI - Mutation A1555G in the 12S rRNA gene and its epidemiological importance in German, Hungarian, and Polish patients. AB - The A1555G mutation in the 12SrRNA gene has been associated with aminoglycoside induced and nonsyndromic sensorineural hearing impairment. In this study we analyzed Hungarian, Polish and German patients with nonsyndromic severe to profound hearing impairment of unknown origin for this mutation. The frequency of the A1555G mutation in the Hungarian hearing impaired population was below 1.8 %. Three out of 125 Polish patients carrying the A1555G mutation were identified. Among German patients one carrier was found (0.7 %) revealing a homoplastic A1555G mutation, whereas no mutation was detected in control individuals with normal hearing (frequency < 0.6%). In summary the frequencies of the A1555G mutation are low in the hearing impaired as well as in the normal population in Hungary, Poland and Germany. Since the importance of this mutation and its relationship with aminoglycoside exposure is not well understood yet, patients with nonsyndromic hearing impairment should be routinely screened for this mutation to avoid aminoglycoside induced hearing impairment due to increased sensitivity of maternal relatives. PMID- 11857752 TI - NF1 mutations in neurofibromatosis 1 patients with plexiform neurofibromas. AB - Neurofibromatosis 1 (NF1) is an autosomal dominant disorder caused by genetic alterations of the NF1 gene on 17q11.2. About 30% of NF1 patients develop plexiform neurofibromas (PNFs), which often cause severe clinical deficits. To determine whether there is a certain genotype underlying PNFs or subtypes of PNFs, we screened 42 NF1 patients from 41 families with PNFs for mutations in the NF1 gene. In 33 out of the 41 (80%) unrelated patients NF1 mutations were found, 24 are novel while the other 9 have been described in previous studies. The 33 mutations included 23 nonsense and frameshift, six splice and four missense mutations. The tumors in these patients had various sizes and features/growth characteristics. No correlation was found between the type or location of the NF1 mutations and size, location or feature of the PNFs, suggesting that many types of NF1 mutations can lead to development of PNFs. PMID- 11857753 TI - Co-existence of frataxin and cardiac troponin T gene mutations in a child with Friedreich Ataxia and familial hypertrophic cardiomyopathy. AB - Friedreich Ataxia (FA) is a neurodegenerative disorder characterised by progressive gait disturbance, dysarthria, dysmetria and other coordination disorders. The genetic defect is represented by an expansion of GAA repeats in the frataxin gene (FRDA or X25). Hypertrophic cardiomyopathy is a common finding in FA, and it is widely recognised as specific for the diagnosis of disease status. In this study, we report the co-existence, in a 5-year old boy with FA, of a double mutation in two distinct genes [X25 (A allele: 850 triplets; B allele: 1000 triplets), and cardiac troponin T (TNNT2) (287G>A)]. TNNT2 gene mutations have been previously identified in individuals with a familial form of hypertrophic cardiomyopathy (FHC), an autosomal dominant inherited disease characterised by unexplained cardiac hypertrophy and high incidence of sudden death. Although we cannot rule out the impact of each gene defect on cardiac morphology, it is of interest that these two mechanisms may be acting in a synergistic fashion to produce the extreme degree of cardiac hypertrophy detected in the child. This is, to our knowledge, the first description of a double gene defect in individuals with FA and FHC. PMID- 11857754 TI - Molecular study of the hydroxymethylbilane synthase gene (HMBS) among Polish patients with acute intermittent porphyria. AB - Acute intermittent porphyria (AIP), an autosomal dominant disorder of heme biosynthesis, is due to mutations in hydroxymethylbilane synthase (HMBS; or porphobilinogen deaminase, PBGD) gene. In this study, we analyzed 20 Polish patients affected by AIP and we were able to characterize seven novel mutations. A nonsense mutation (Y46X), two frameshift mutations (315delT and 552delT) and a 131bp deletion (nucleotides 992-1123) give rise to truncated proteins. A donor splice site mutation IVS12+2T>C predicts skipping of exon 12. A missense mutation (D61Y) was identified in two apparently unrelated patients with a clearly clinical indication of AIP. An inframe 3-bp deletion (278-280delTTG) results in the removal of V93 from the enzyme. In addition to the novel mutations, nine previously described HMBS gene mutations-R26H, G111R, IVS7+1G>A, R149X, R173Q, 730-731delCT, R225X, 982-983delCA and G335D-were identified in this cohort. Our results demonstrate that molecular analysis of the PBGD gene is a more reliable method comparing to enzymatic assay in the diagnosis of AIP. Although more than 170 different mutations are known to the HMBS gene so far, over 40% of all mutations identified among the Polish AIP patients of this study are novel mutations, indicating the heterogeneity of molecular defects causing AIP. PMID- 11857755 TI - Comparison of SSCP and DHPLC for the detection of LDLR mutations in a New Zealand cohort. AB - Familial hypercholesterolaemia (FH) is a common inherited disorder, associated with premature vascular disease. FH may be caused by many different mutations in the low density lipoprotein receptor (LDLR) gene, about 700 mutations have been described, most of which occur rarely and often only in single families. Although particular mutations are prevalent in certain ethnic groups, countries with heterogeneous population bases (such as NZ) may carry a wide variety of mutations; making a gene screening approach the appropriate first step for a mutation detection programme. We have compared SSCP with DHPLC to assess their effectiveness as methods for LDLR mutation detection. Although five novel LDLR mutations were detected by SSCP in patients with FH, DHPLC was more sensitive, with eight novel mutations detected. Six of these mutations (T392M, R419G, Y421N, 1206-1207delCT, 1872delC, and 1943delC) were clustered in exons 9 and 13 of the EGF precursor homology domain, one (679-680delAC) in the ligand binding domain (exon 4) and the eighth (P774H) in the membrane-spanning domain (exon 16). Twenty five mutations were identified in 35 patients in total. Of these, we were able to detect only 64% of mutations by SSCP even though all variants were detected by DHPLC. All patients are heterozygous for the mutations, which is consistent with the clinical phenotypes. PMID- 11857757 TI - Comparative proteomics based on stable isotope labeling and affinity selection. AB - Disease, external stimuli (such as drugs and toxins), and mutations cause changes in the rate of protein synthesis, post-translational modification, inter compartmental transport, and degradation of proteins in living systems. Recognizing and identifying the small number of proteins involved is complicated by the complexity of biological extracts and the fact that post-translational alterations of proteins can occur at many sites in multiple ways. It is shown here that a variety of new tools and methods based on internal standard technology are now being developed to code globally all peptides in control and experimental samples for quantification. The great advantage of these stable isotope-labeling strategies is that mass spectrometers can rapidly target those proteins that have changed in concentration for further analysis. When coupled to stable isotope quantification, targeting can be further focused through chromatographic selection of peptide classes on the basis of specific structural features. Targeting structural features is particularly useful when they are unique to types of regulation or disease. Differential displays of targeted peptides show that stimulus-specific markers are relatively easy to identify and will probably be diagnostically valuable tools. PMID- 11857758 TI - Mono and double polar [4 + 2(+)] Diels-Alder cycloaddition of acylium ions with O heterodienes. AB - Gas-phase reactions of acylium ions with alpha,beta-unsaturated carbonyl compounds were investigated using pentaquadrupole multiple-stage mass spectrometry. With acrolein and metacrolein, CH(3)-C(+)(double bond)O, CH(2)(double bond)CH-C(+)(double bond)O, C(6)H(5)-C(+)(double bond)O, and (CH(3))(2)N-C(+)(double bond)O react to variable extents by mono and double polar [4 + 2(+)] Diels-Alder cycloaddition. With ethyl vinyl ketone, CH(3)-C(+)(double bond)O reacts exclusively by proton transfer and C(6)H(5)-C(+)(double bond)O forms only the mono cycloadduct whereas CH(2)(double bond)CH-C(+)(double bond)O and (CH(3))(2)N-C(+)(double bond)O reacts to great extents by mono and double cycloaddition. The positively charged acylium ions are activated O heterodienophiles, and mono cycloaddition occurs readily across their C(+)(double bond)O bonds to form resonance-stabilized 1,3-dioxinylium ions which, upon collisional activation, dissociate predominantly by retro-addition. The mono cycloadducts are also dienophiles activated by resonance-stabilized and chemically inert 1,3-dioxonium ion groups, hence they undergo a second cycloaddition across their polarized C(double bond)C ring double bonds. (18)O labeling and characteristic dissociations displayed by the double cycloadducts indicate the site and regioselectivity of double cycloaddition, which are corroborated by Becke3LYP/6-311++G(d,p) calculations. Most double cycloadducts dissociate by the loss of a RCO(2)COR(1) molecule and by a pathway that reforms the acylium ion directly. The double cycloadduct of the thioacylium ion (CH(3))(2)N-C(+)(double bond)S with acrolein dissociates to (CH(3))(2)N C(+)(double bond)O in a sulfur-by-oxygen replacement process intermediated by the cyclic monoadduct. The double cycloaddition can be viewed as a charge-remote type of polar [4 + 2(+)] Diels-Alder cycloaddition reaction. PMID- 11857759 TI - Gas-phase ion chemistry in the ternary SiH(4)-C(3)H(6)-PH(3) system. AB - Propene-phosphine and the silane-propene-phosphine gaseous mixtures were studied by ion trap mass spectrometry. For the binary mixture the variation of ion abundances under different partial pressures and the mechanisms of ion-molecule reactions are reported. Moreover, the rate constants of the main processes were measured and compared with the collisional rate constants to determine the reaction efficiencies. In the ternary silane-propene-phosphine mixture the mechanisms of formation of Si(m)C(n)P(p)H(+)(s) ion clusters were elucidated, but the complexity of the system and the low abundances of the ions usually isolated by successive steps prevented the determination of rate constants. The hydrogenated ternary ions are mainly formed by reactions of Si(r)P(s)H(+)(t) ions with propene, whereas a minor contribution comes from reactions of Si(m)C(n)H(+)(p) ions with phosphine. The C(v)P(w)H(+)(z) ions show very low reactivity with silane. The formation processes of these species are discussed in relation to their possible role as precursors of amorphous silicon carbides doped with phosphorus obtained by deposition from properly activated silane-propene phosphine mixtures. PMID- 11857760 TI - Structurally diagnostic ion-molecule reactions: acylium ions with alpha-, beta- and gamma-hydroxy ketones. AB - Gas-phase reactions of four acylium ions and a thioacylium ion with three isomeric alpha-, beta- and gamma-hydroxy ketones are performed by pentaquadrupole mass spectrometric experiments. Novel structurally diagnostic reactions are observed, and found to correlate directly with interfunctional group separation. All five ions tested (CH(3)CO(+), CH(2)(double bond)CHCO(+), PhCO(+), (CH(3))(2)NCO(+) and (CH(3))(2)NCS(+)) react with the gamma-hydroxy ketone (5 hydroxy-2-pentanone) to form nearly exclusively a cyclic oxonium ion of m/z 85 that formally arises from hydroxy anion abstraction. With the beta-hydroxy ketone (4-hydroxy-2-pentanone), CH(2)(double bond)CHCO(+), PhCO(+) and (CH(3))(2)NCO(+) form adducts that undergo fast cyclization via intramolecular water displacement, yielding resonance-stabilized cyclic dioxinylium ions. With the alpha-hydroxy ketone (3-hydroxy-3-methyl-2-butanone), PhCO(+), (CH(3))(2)NCO(+) and (CH(3))(2)NCS(+) form stable adducts. Evidence that these adducts display cyclic structures is provided by the triple-stage mass spectra of the (CH(3))(2)NCS(+) adduct; it dissociates to (CH(3))(2)NCO(+) via a characteristic reaction dissociation pathway that promotes sulfur-by-oxygen replacement. If cyclizations are assumed to occur with intramolecular anchimeric assistance, relationships between structure and reactivity are easily recognized. PMID- 11857761 TI - Mass spectrometric studies of 2-aryl-5-acetylthiazole derivatives. AB - Electron ionisation mass spectrometry was usefully used to characterize structurally 2-aryl-5-acetylthiazole derivatives in the gas phase. The compounds show characteristic fragmentation pathways depending on the chemical nature of the substituent at position 2, consisting mainly in the cleavage of both the 1,2- and 3,4-bonds of the thiazole ring. Liquid secondary ion mass spectrometry was applied to study the effects of protonation on the gas-phase unimolecular reactions of this class of compound. Tandem mass spectrometric experiments, carried out on molecular and protonated molecular ions, and also on fragment ions produced in the source, allowed the elucidation of gas-phase decompositions of low-internal energy ions. PMID- 11857762 TI - Phosphopeptide detection and sequencing by matrix-assisted laser desorption/ionization quadrupole time-of-flight tandem mass spectrometry. AB - A prototype matrix-assisted laser desorption/ionization quadrupole time-of-flight (MALDI-TOF) tandem mass spectrometer was used to sequence a series of phosphotyrosine-, phosphothreonine- and phosphoserine-containing peptides. The high mass resolution and mass accuracy of the instrument allowed the localization of one, three or four phosphorylated amino acid residues in phosphopeptides up to 3.1 kDa. Tandem mass spectra of two different phosphotyrosine peptides permitted amino acid sequence determination and localization of one and three phosphorylation sites, respectively. The phosphotyrosine immonium ion at m/z 216.04 was observed in these MALDI low-energy CID tandem mass spectra. Elimination of phosphate groups was evident from the triphosphorylated peptide but not from the monophosphorylated species. The main fragmentation pathway for the synthetic phosphothreonine-containing peptide and for phosphoserine containing peptides derived from beta-casein and ovalbumin was the beta elimination of phosphoric acid with concomitant conversion of phosphoserine to dehydroalanine and phosphothreonine to 2-aminodehydrobutyric acid. Peptide fragment ions of the b- and y-type allowed, in all cases, the localization of phosphorylation sites. Ion signals corresponding to (b-17), (b-18) and (y-17) fragment ions were also observed. The abundant neutral loss of phosphoric acid ( 98 Da) is useful for femtomole level detection of phosphoserine-peptides in crude peptide mixtures generated by gel in situ digestion of phosphoproteins. PMID- 11857763 TI - Uncoupled analysis of secondary and tertiary protein structure by circular dichroism and electrospray ionization mass spectrometry. AB - Electrospray ionization mass spectrometry (ESI-MS) applied to protein conformational studies is a powerful new method that seems to provide specific information about protein tertiary structure. In this study, we analyzed the effect of trifluoroethanol (TFE) on a myoglobin peptide and cytochrome c (cyt c) at low pH by circular dichroism (CD) and ESI-MS. These experiments show that coil to-helix transition per se does not affect ESI mass spectra, confirming that this technique is insensitive to the local conformation of the polypeptidic chain and, rather, reports on the tertiary contacts characterizing different protein conformations. This property makes ESI-MS an excellent method, complementary to CD, for the characterization of protein conformational changes. Fluorinated alcohols have been suggested to induce molten globule formation in acid-unfolded cyt c. The experiments described here show that TFE does not induce major changes in the ESI mass spectrum of cyt c at pH 2.2, indicating that no stabilization of compact, globular structures is detectable under the conditions employed. On the other hand, even low concentrations of TFE (2-5%) are shown to destabilize the folded state of the protein around the mid-point of its acid-induced unfolding transition. PMID- 11857764 TI - Electrospray ionization mass spectrometry and tandem mass spectrometry of clodronate and related bisphosphonate and phosphonate compounds. AB - The bisphosphonate family with a P-C-P structure is a broad class of drugs, widely investigated as potential inhibitors in bone diseases and calcium metabolic disorders. In this study, the mass spectrometric (MS) behavior and fragmentation of clodronate and related bisphosphonate and phosphonate compounds was studied by using negative ion electrospray ionization (ESI) with triple quadrupole and ion trap instruments. The effect of pH on the degree of deprotonation of the polyprotic bisphosphonic and phosphonic acids in negative ion ESI-MS was investigated, and the degree of deprotonation in the ESI mass spectra and the dissociation in the liquid phase were compared. The results provide evidence that the measured ESI mass spectra do not correlate with the chemistry in the liquid phase owing to the decrease in the pH of the solvent droplets during the ion evaporation process and the charge state neutralization in the gas phase. Ion trap MS(n) provided useful information on the fragmentation study of clodronate and related bisphosphonate and phosphonate compounds, in which interesting fragmentation pathways including the direct elimination of carbon monoxide from deprotonated bisphosphonates and formation of a P-P bond were observed. Reactions between the product ions with a -PO(2) group and residual water in the ion trap or in the high-pressure region of the triple quadrupole instrument formed other unexpected fragmentation paths for all the bisphosphonates studied. PMID- 11857765 TI - Facile distinction of neutral and acidic tetraether lipids in archaea membrane by halogen atom adduct ions in electrospray ionization mass spectrometry. AB - Calditocaldarchaeol (neutral tetraether lipid) from Sulfolobus acidocaldarius (acidothermophilic archaea) and intact total lipid from the thermoacidophilic archaea Sulfolobus sp. was examined by electrospray ionization time-of-flight mass spectrometry in the negative-ion mode using high resolution. When the sample was injected as a solution in a 3:1 mixture of methanol (MeOH) and chloroform (CHCl(3)) using an infusion system, the total ether lipid afforded molecular related ions as [M - H](-) for acidic polar lipids containing a phosphoric or sulfuric group, and as [M + Cl](-) ion for neutral glycolipids. The attachment of chloride was confirmed by the observation of [M + Br](-) ion, instead of [M + Cl](-) ion, when a 3:1 mixture of MeOH and CHBr(3) was used in place of MeOH CHCl(3) as the solvent. The composition of tetraether neutral glycolipids that are different from each other only in the number of five-membered rings in the isoprenoid chain was determined on the basis of the isotope-resolved mass spectrum of [M + Cl](-) ions. As for acidic tetraether lipids, molecular-related ions [M - H](-)) were not observed when the 3:1 MeOH-CHBr(3) mixture was used as the solvent. These results together afforded a facile method of distinguishing neutral from acidic tetraether lipids in intact total lipids of acidothermophilic archaea. This method was applied to determine the difference of the number of five-membered rings in isoprenyl chains of neutral tetraether glycolipids yielded by the Sulfolobus sp. grown at different temperatures. Discrimination of neutral tetraether glycolipids from acidic tetraether lipids in the total lipids obtained from Thermoplasma sp. was also achieved by this method. PMID- 11857766 TI - Liquid chromatographic/mass spectrometric determination of biologically active alkaloids in extracts of Peschiera fuschiaefolia. AB - Four alkaloids were isolated from an alcoholic extract of the bark from the stem of Peschiera fuschiaefolia. Two of these compounds, voacamine and voacamidine, are dimeric alkaloids which are thought to be responsible for the antimalarial activity of these extracts. The mass spectra and the response factors of these four compounds were obtained by liquid chromatography/mass spectrometry in the electrospray positive ionization mode. The concentrations of these alkaloids were measured in two different P. fuschiaefolia extracts. The ion chromatograms of the two extracts were compared on the basis of their [M + H](+) and [M + 2H](+2) ions characteristic of various alkaloids previously isolated from P. fuschiaefolia bark. The two extracts were found to differ mostly in the relative concentrations of the dimeric alkaloids. PMID- 11857767 TI - Automated deconvolution and deisotoping of electrospray mass spectra. AB - Electrospray ionization (ESI) of peptides and proteins produces a series of multiply charged ions with a mass/charge (m/z) ratio between 500 and 2000. The resulting mass spectra are crowded by these multiple charge values for each molecular mass and an isotopic cluster for each nominal m/z value. Here, we report a new algorithm simultaneously to deconvolute and deisotope ESI mass spectra from complex peptide samples based on their mass-dependent isotopic mean pattern. All signals corresponding to one peptide in the sample were reduced to one singly charged monoisotopic peak, thereby significantly reducing the number of signals, increasing the signal intensity and improving the signal-to-noise ratio. The mass list produced could be used directly for database searching. The developed algorithm also simplified interpretation of fragment ion spectra of multiply charged parent ions. PMID- 11857768 TI - Application of a portable quadrupole mass spectrometer in Paks Nuclear Power Plant for control of steam generator chemical cleaning. PMID- 11857770 TI - Current literature in mass spectrometry. PMID- 11857771 TI - Depletion of mitochondrial DNA and enzyme in estrogen-induced hamster kidney tumors: a rodent model of hormonal carcinogenesis. AB - Mitochondrial DNA (mtDNA) encodes for 13 polypeptides critical for normal functioning of the electron transport chain and damage to mtDNA has been associated with aging, and implicated in several disease processes. Although damage to mtDNA is being implicated in mutagenesis and carcinogenesis, there are limited studies demonstrating the role and extent of mtDNA damage in human or rodent cancers. Using serial dilution and competitive polymerase chain reaction analysis, we have quantitated the amount of total mtDNA and analyzed the extent of mtDNA damage in estrogen-induced and estrogen-dependent hamster kidney tumors. The hamster kidney tumor model is a useful and widely investigated rodent model of hormonal carcinogenesis, which shares several characteristics with human breast and uterine cancers, and point to a common mechanistic pathway. Our data indicate a significant decrease in the copy number of total mtDNA and the activity of a nuclear-encoded mitochondrial enzyme citrate synthase in hamster kidney tumors compared to age-matched controls. Since there are several hundred mitochondria in a cell and each mitochondrion has multiple copies of mtDNA, a very small percentage of somatic deletion mutation may not be enough to result in a decreased capacity of the mitochondrial genome. However, a significant increase in deletion mutations or a decrease in the mtDNA copy number can result in a decreased oxidative phosphorylation capacity of the mitochondria and decreased energetics, and thus increased susceptibility to the disease process. Therefore, estrogen-induced hamster kidney tumor model can be a useful rodent model of carcinogenesis to understand the role of mtDNA damage in cancer progression and development. PMID- 11857772 TI - Activation of a stress-induced gene by insecticides in the midge, Chironomus yoshimatsui. AB - Stress proteins (heat shock proteins, HSPs) have been proposed as general biomarkers for environmental monitoring. In the present study, we evaluated the environmental stress-burden on the aquatic midge Chironomus yoshimatsui using hsp70 expression. Larvae collected from streams receiving polluted runoff (field strain) were resistant to the organophosphorus insecticide, fenitrothion (F), and the synthetic pyrethroid, ethofenprox (E), whereas a strain originally collected from an unpolluted area (susceptible strain) showed low resistance to insecticide exposure. To examine the expression of an HSP70 gene in C. yoshimatsui, an hsp70 cDNA probe was prepared using RNA obtained from the field strain larvae and used for Northern blot analyses. The expression of this HSP70 gene in larvae collected from two field sites in May about 1 week after insecticide spraying in the fields was 2.3 (p = 0.018) to 3.3 fold higher than that in the susceptible strain and was also 4.6 and 1.4 (p = 0.033) fold higher than those collected in November 3 months after the cessation of insecticide spraying. In order to identify potential inducers of the HSP70 gene of the field strain, larvae of the susceptible strain were exposed to F or E for 24 h and hsp70 mRNA levels determined. Exposures to F at 0.4 microg/L and E at 1.1 microg/L increased hsp70 mRNA levels 2.7 (p = 0.049) and 4.4 (p = 0.043) fold over controls, respectively. These results suggest that larvae collected from polluted areas are burdened by environmental stressors and the tested insecticides are potential inducers of HSP70. The results also support the suggestion that HSP70 gene expression is a sensitive indicator of low level (nonlethal) exposures to certain insecticides. PMID- 11857773 TI - Inhibition of human squalene monooxygenase by selenium compounds. AB - Selenosis in animals is characterized by a variety of neurological abnormalities, but the chemical species of selenium and the molecular targets that mediate this neurotoxicity are unknown. We have previously shown that selenite is a potent inhibitor of squalene monooxygenase, the second enzyme in the committed pathway for cholesterol biosynthesis; inhibition of this enzyme by dimethyltellurium leads to a peripheral demyelinating neuropathy similar to that seen in selenosis. To evaluate the role methylation plays in selenium toxicity, we examined the ability of three methylselenium compounds, methylselenol, dimethylselenide, and trimethylselenonium iodide, to inhibit purified recombinant human squalene monooxygenase. IC(50) values for methylselenol (95 microM) and dimethylselenide (680 microM) were greater than that previously obtained for selenite (37 microM), and inhibition by trimethylselenonium iodide was evident only at concentrations above 3 mM. Inhibition by methylselenol as well as by selenite was slow and irreversible, suggestive of covalent binding to the enzyme, and thiol-containing compounds could prevent and reverse this inhibition, indicating that these compounds were reacting with sulfhydryl groups on the protein. Monothiols such as glutathione and beta-mercaptoethanol provided better protection than did dithiols, suggesting that these selenium compounds bind to only one of the two proposed vicinal cysteines on squalene monooxygenase. Unexpectedly, the inhibition by selenite was significantly enhanced by dithiols, indicating that a more toxic species, possibly selenide, was formed in the presence of these dithiol reductants. PMID- 11857774 TI - A comparison of 60, 70, and 90 kDa stress protein expression in normal rat NRK-52 and human HK-2 kidney cell lines following in vitro exposure to arsenite and cadmium alone or in combination. AB - Arsenite and cadmium are two potent nephrotoxicants and common Superfund site elements. These elements are included among the stress protein inducers, but information regarding relationships between toxicity produced by combinations of these agents to the stress protein response is lacking. In this study, the immortalized cell lines normal rat kidney NRK-52E and human kidney HK-2 were exposed in vitro to arsenite (As(3+)), cadmium (Cd(2+)), or to equimolar As(3+) plus Cd(2+) mixture combinations for 3 and 5 h over a concentration range of 0.1 100 microM. After a 12-h recovery period, cultured cells were then evaluated for expression of the 60, 70, and 90 kDa major stress protein families. Results indicated that expression of stress proteins varied depending on the species of kidney cells exposed, the exposure concentrations, and the length of exposure to each element on an individual basis and for combined mixtures. For the HK-2 kidney cell line, increased levels of the 70 kDa stress protein was observed for single and combined element exposures whereas there was no change or a decrease of stress proteins 60 and 90 kDa. Increased 70 kDa expression was observed for 10 microM doses of single elements and for a lower dose of 1 microM of the As plus Cd mixture at 3- and 5-h exposures. NRK-52 kidney cells exposed to equivalent doses of As(3+) and Cd(2+) alone or in combination showed increased levels of all stress proteins 60, 70, and 90 kDa. This increase was seen for 10 microM of the As plus Cd mixture at 3 h whereas for single element exposures, increased stress protein levels were generally observed for the 100-microM doses. At 5 h- exposure, 60 and 90 kDa levels increased for 10 microM of Cd(2+) and 60 kDa levels increased for 1 microM of As(3+). However, exposures to 10 microM of the As plus Cd mixture decreased 60 kDa protein expression to control levels at 5 h. For both kidney cell lines, there was a decrease in the stress protein expression levels for all three stress protein families for 100-microM doses of the mixture combination for 3- and 5-h exposures. These data indicate a dose- and combination related correlation between depression of the stress protein response and the onset of overt cellular toxicity and/or cell death. The threshold for these changes was cell line specific. PMID- 11857775 TI - Selective induction of mitogen-activated protein kinases in human lens epithelial cells by ultraviolet radiation. AB - The present study investigates the effects of ultraviolet radiation (UVR) on mitogen-activated protein kinase (MAPK) activity in human lens epithelial (HLE) cells. Irradiation of HLE cells with ultraviolet B and ultraviolet C radiation activates the stress-response MAPK proteins, p38 and c-Jun NH(2)-terminal kinase (JNK), in a dose- and time-dependent manner, while the extracellular-regulated signal kinase (ERK 44/42) cascade was not altered by UVR exposure. Ultraviolet A radiation failed to elicit a MAPK response. UVR-induced MAPK activation does not require protein kinase C or phosphatidylinositol 3-kinase activity, suggesting that this is not a receptor-mediated event. Inhibition of ribosomal translation completely abolished UVR-induced MAPK activation, while treatment with the antioxidant, N-acetyl cysteine, and mild heat shock had no effect on this activation. These data demonstrate for the first time the selective activation of MAPK cascades in a lens epithelial cell line. PMID- 11857776 TI - Effect of peroxisome proliferators on the methylation and protein level of the c myc protooncogene in B6C3F1 mice liver. AB - Peroxisome proliferators in general are nongenotoxic mouse liver carcinogens for which DNA hypomethylation and altered gene expression are proposed mechanisms. Therefore, the peroxisome proliferators 2,4-dichlorophenoxyacetic acid (2,4-D), dibutyl phthalate (DBP), gemfibrozil, and Wy-14,643 were evaluated for the ability to alter the methylation and expression of the c-myc protooncogene. Male B6C3F1 mice were administered for 6 days in their diet Wy-14,643 (5-500 ppm), 2,4 D (1,680 ppm), DBP (20,000 ppm), or gemfibrozil (8,000 ppm). All four peroxisome proliferators caused hypomethylation of the c-myc gene in the liver. Wy-14,643 appeared to be the most efficacious with a threshold between 10 and 50 ppm. The level of the c-myc protein was increased by Wy-14,643, but not the other peroxisome proliferators. When female B6C3F1 mice received a two-thirds partially hepatectomy and 16 h later were administered 50 mg/kg Wy-14,643 by gavage, hypomethylation of the gene occurred 24 h later. Hypomethylation was not found in mice that received Wy-14,643 following a sham operation. Hypomethylation of the c myc gene within 24 h of administering Wy-14,643 after a partial hepatectomy but not after a sham operation supports the hypothesis that the peroxisome proliferators prevent methylation of hemimethylated sites formed by DNA replication. PMID- 11857777 TI - Conditional alleles in mice: practical considerations for tissue-specific knockouts. PMID- 11857778 TI - Corralling conditional mutations: a unified resource for mouse phenotypes. PMID- 11857779 TI - Generation of a loxP flanked bmp4loxP-lacZ allele marked by conditional lacZ expression. PMID- 11857780 TI - Generation of Bmpr/Alk3 conditional knockout mice. PMID- 11857781 TI - Conditional inactivation of the TGF-beta type II receptor using Cre:Lox. PMID- 11857782 TI - Conditional knockout of the Smad1 gene. PMID- 11857783 TI - Generation of Smad4/Dpc4 conditional knockout mice. PMID- 11857784 TI - Generation of the floxed allele of the SIP1 (Smad-interacting protein 1) gene for Cre-mediated conditional knockout in the mouse. PMID- 11857785 TI - Generation of Fgfr1 conditional knockout mice. PMID- 11857786 TI - A conditional floxed (loxP-flanked) allele for the retinoic acid receptor alpha (RARalpha) gene. PMID- 11857787 TI - A conditional floxed (loxP-flanked) allele for the retinoic acid receptor beta (RARbeta) gene. PMID- 11857789 TI - AF-2 knock-in mutation of estrogen receptor alpha: Cre-loxP excision of a PGK-neo cassette from the 3' UTR. PMID- 11857788 TI - A conditional floxed (loxP-flanked) allele for the retinoic acid receptor gamma (RARgamma) gene. PMID- 11857790 TI - Conditional inactivation of the dopamine receptor 5 gene: flanking the Drd5 gene with loxP sites. PMID- 11857791 TI - A Hoxa2 mutant conditional allele generated by Flp- and Cre-mediated recombination. PMID- 11857792 TI - A Hoxa2 knockin allele that expresses EGFP upon conditional Cre-mediated recombination. PMID- 11857793 TI - Conditional, floxed allele of the Krox20 gene. PMID- 11857794 TI - Generation of a conditional loxP allele of the Pax3 transcription factor that enables selective deletion of the homeodomain. PMID- 11857795 TI - Conditional inactivation of Lim1 function. PMID- 11857796 TI - Conditional inactivation of Sox9: a mouse model for campomelic dysplasia. PMID- 11857797 TI - Generation of mice carrying conditional knockout alleles for the transcription factor SRF. PMID- 11857798 TI - Conditional inactivation of transcription factor AP-2gamma by using the Cre/loxP recombination system. PMID- 11857799 TI - Generation of a conditionally null allele of hnf4alpha. PMID- 11857800 TI - Generation and functional confirmation of a conditional null PPARgamma allele in mice. PMID- 11857801 TI - Conditional inactivation of p63 by Cre-mediated excision. PMID- 11857802 TI - Generation of an Mdm2 conditional allele in mice. PMID- 11857803 TI - Conditional allele of mdm2 which encodes a p53 inhibitor. PMID- 11857804 TI - Cre/loxP-mediated inactivation of the murine Pten tumor suppressor gene. PMID- 11857805 TI - Targeting and conditional inactivation of the murine Men1 locus using the Cre recombinase: loxP system. PMID- 11857806 TI - A conditional null allele of the major histocompatibility IA-beta chain gene. PMID- 11857807 TI - Cre recombinase-controlled expression of the mb-1 allele. PMID- 11857808 TI - Construction of a conditional allele of RSK-B/MSK2 in the mouse. PMID- 11857809 TI - Conditional mutagenesis of CamKIV. PMID- 11857810 TI - Conditional inactivation of the calbindin D-28k (Calb1) gene by Cre/loxP-mediated recombination. PMID- 11857811 TI - Conditional gene targeting of the Scnn1a (alphaENaC) gene locus. PMID- 11857812 TI - A conditional allele at the mouse channel activating protease 1 (Prss8) gene locus. PMID- 11857813 TI - Generation of Pex5-loxP mice allowing the conditional elimination of peroxisomes. PMID- 11857814 TI - Conditional inactivation of the peroxisome biogenesis Pex13 gene by Cre-loxP excision. PMID- 11857816 TI - Flp recombinase regulated lacZ expression at the ROSA26 locus. PMID- 11857817 TI - A targeted X-linked CMV-Cre line. PMID- 11857815 TI - Mice with Cre recombinase activatable PDGF-C expression. PMID- 11857818 TI - K14-GLp65 transactivator induces transgene expression in embryonic epidermis. PMID- 11857819 TI - In utero activation of K5.CrePR1 induces gene deletion. PMID- 11857820 TI - The UTHSCSA Dental School in the new millennium. AB - The UTHSCSA Dental School, just short of 30 years in existence, has made great strides to be a leader in dental education. Although increased use of computers has the greatest potential for revolutionizing dental education, there are other components that must constantly be evaluated and improved. A major curriculum review is in progress. The process is a grass roots effort to allow input from faculty, students, alumni and outside consultants. The school's competencies are being reevaluated to assure they are contemporary and the methods to assess them are valid and reliable. The way we evaluate applicants is an ongoing evolution. Our appropriate role in the community (local, national, and international) continues to be a challenge. Success will be measured by how well we continuously evaluate our mission and goals, identify problems and find and implement solutions. PMID- 11857821 TI - Baylor College of Dentistry welcomes the new millennium. PMID- 11857822 TI - Oral and maxillofacial pathology case of the month. Recurrent aphthous stomatitis -herpetiform type. PMID- 11857823 TI - High noon on patient protection approaches in Washington. PMID- 11857824 TI - Oral and maxillofacial pathology case of the month. Pernicious anemia. PMID- 11857825 TI - Protecting your employees and patients against latex allergy. PMID- 11857826 TI - Infancy ideal time to begin prevention of dental disease. PMID- 11857827 TI - Address to the House of Delegates May 6, 1999. PMID- 11857828 TI - Oral and maxillofacial pathology case of the month. Focal cemento-osseous dysplasia. PMID- 11857829 TI - How do patients judge a good dentist? PMID- 11857830 TI - Offering additional payment options for increasing referrals. PMID- 11857831 TI - Direct reimbursement finds success on the south plains. PMID- 11857832 TI - Texas Dental Association 1998 financial report. PMID- 11857833 TI - Forensic dentistry. Law enforcement seeks TDA's help. PMID- 11857834 TI - Questions and answers about dental management service organizations [Interview by Arthur H. Jeske]. PMID- 11857835 TI - Oral and maxillofacial pathology case of the month. Chondrosarcoma grade II. PMID- 11857836 TI - Give me liberty? Give me direct reimbursement. PMID- 11857837 TI - Chairside glucose testing: an inexpensive, invaluable diagnostic tool for dental offices. PMID- 11857838 TI - Overview of American Heart Association Protocols. Part 1--B.L.S. PMID- 11857839 TI - Oral and maxillofacial pathology case of the month. Actinomycosis. PMID- 11857840 TI - This could never happen to me! White-collar crime in the dental office. PMID- 11857841 TI - TDA membership: what is it worth to you? PMID- 11857842 TI - Oral and maxillofacial pathology case of the month. Peripheral odontogenic (ossifying) fibroma. PMID- 11857843 TI - Membership: the association bedrock. PMID- 11857844 TI - Meeting challenges together. PMID- 11857845 TI - Dental fluorosis among third graders in Harris County, Texas--1998 study findings. PMID- 11857847 TI - It's in our hands--so, get involved! PMID- 11857848 TI - The Dunn light. PMID- 11857846 TI - New drugs and product approvals from 1999. PMID- 11857849 TI - Oral health care in Texas. PMID- 11857850 TI - Oral and maxillofacial pathology case of the month. Central giant cell granuloma. PMID- 11857851 TI - Clinical warning signs and symptoms of head and neck cancer. PMID- 11857852 TI - The oral brush biopsy: it's as easy as 1, 2, 3. PMID- 11857853 TI - Treatment of nicotine addiction. AB - The best available data show smoking to be by far the most important cause of disease and death in our society, contributing to an average of 1000 deaths every day. Although a large majority of current smokers express a desire to quit, the majority of "self-help" attempts to quit are not successful. Further, most smokers indicate never having received advice on cessation from healthcare providers. The combination of pharmacotherapy and behavioral interventions, even on a minimal level, have been shown to be effective in cessation. Such strategies are certainly highly cost-effective, given the enormous costs to society of smoking-related illnesses. Therefore, it is important that all healthcare providers provide at least some form of smoking cessation programs for their patients. PMID- 11857854 TI - The role of dentistry in palliative care of the head and neck cancer patient. AB - Although palliative care for the terminally ill is based on a multidimensional philosophy to provide whole-person comfort care while maintaining optimal function, it does not usually include dentistry in its team approach. Dentists can have a significant role in the care of these patients by providing total, active comfort care of the oral cavity. The function of the oral cavity is essential to the patient's ability to thrive. Therefore, alleviation of pain and prevention of infection in the oral cavity should be a priority in providing total, active comfort for the patient. The oral problems experienced by the hospice head and neck patient clearly affect the quality of his or her remaining life. Through routine assessments and interventions by a dentist on the palliative care team (Figure 4), comfort care for the patient may be improved by the maintenance of oral hygiene and procedures to hydrate the oral mucosa. In addition, routine dental assessments may identify dental disease and facilitate dental interventions for caries, periodontal disease, oral mucosal problems or prosthetic needs. Attention to such detail may reduce not only the microbial load of the mouth but the risk for pain and oral infection as well. This multidisciplinary approach to palliative care, including a dentist, may reduce the oral debilities that influence the patient's ability to speak, eat or swallow. Not only does maintenance of oral health have impact on the quality of life, which is already challenged by the disease, but it also aids in the ability of patients to thrive for whatever precious time is left to them. PMID- 11857855 TI - Reimbursement of cancer-related oral care. PMID- 11857857 TI - Trigeminal neuralgia in dentistry. PMID- 11857856 TI - Overview of American Heart Association protocols. Part 2--A.C.L.S. PMID- 11857858 TI - Nine non-clinical issues that increase liability; business practices do affect patient satisfaction. PMID- 11857859 TI - Strategy for implementing new technologies. PMID- 11857860 TI - Oral and maxillofacial pathology case of the month. Lymphangioma. PMID- 11857861 TI - Dentistry in Texas 2010--fact or fiction? PMID- 11857863 TI - Dentistry in the new millennium. PMID- 11857862 TI - The demographic outlook for orthodontists in Texas at the start of the next millennium. AB - Like all health care professionals, orthodontists are concerned about the need and demand for their services. Orthodontists should experience an increase in demand for their services as a result of an increasing population and an aging orthodontist population. How many additional orthodontic graduates do we need per year in Texas? Based on many factors, Texas dental schools could graduate additional students per year to be consistent with those factors. In conclusion, it appears that Texas can easily support three graduate orthodontic residency programs. As we enter the next millennium, the future of orthodontics has never looked better. PMID- 11857864 TI - Dental materials in the new millennium: research at Baylor College of Dentistry and a look at new trends. PMID- 11857865 TI - Pharmacology at the beginning of the 21st century: implications of several new developments for dentistry and current status of the major 20th century dental drugs. AB - This review has highlighted some of the developments in pharmacology likely to impact dentistry as we enter the 21st century. While dentistry will naturally lag behind medicine in the introduction of new drugs, we must constantly explore all information resources available to keep abreast of developments in pharmacology that will affect our patients and our use of drugs in dental practice. This century will abound with new therapies, some of which will hopefully lead us out of our tradition of using rather non-selective, widely distributed chemicals with multiple actions and side effects into an age of highly-selective therapies, including the repair and replacement of defective genetic determinants of disease with healthy ones. PMID- 11857866 TI - Dental office equipment: then, now, & to come. PMID- 11857867 TI - Bacteria in dental operatory water is still a problem. Dentists alerted to clinical, legal implications. PMID- 11857869 TI - State board of dental examiners complaint/investigation process. PMID- 11857868 TI - Oral and maxillofacial pathology case of the month. Primary herpetic gingivostomatitis. PMID- 11857870 TI - Fabrication of a custom grid using orthodontic materials to locate a pellet in a patient's hand. PMID- 11857871 TI - The Jordan family legacy: three generations of dentistry. PMID- 11857872 TI - Heartbreak with a smile. PMID- 11857873 TI - The principles of ethics and code of professional conduct: what does it mean to you? PMID- 11857874 TI - Hands-on volunteerism. PMID- 11857875 TI - So it's time to have your X-ray unit inspected! PMID- 11857877 TI - The time has come. PMID- 11857876 TI - Oral and maxillofacial pathology case of the month. Myospherulosis. PMID- 11857878 TI - New specialty recognized: oral and maxillofacial radiology. PMID- 11857879 TI - Oral and maxillofacial pathology case of the month. Condyloma acuminatum (venereal wart). PMID- 11857880 TI - Questions and answers about long-term care insurance. PMID- 11857881 TI - Child abuse: an overview. PMID- 11857882 TI - Recognizing and reporting the orofacial trauma of child abuse/neglect. PMID- 11857884 TI - Domestic violence: identifying the deadly silence. AB - In the past decade, domestic violence, or intimate partner violence (IPV), has become recognized as a serious public health issue in the United States (2, 13). This devastating abuse affects 3 to 4 million individuals every year in this country alone (6, 7, 14). Millions more are affected due to the children and family who witness the abuse. Each year 3.3 million children witness domestic partner abuse and 50 to 70 percent of the perpetrators also abuse their children (7, 1). Domestic violence is a leading cause of serious injury to women, killing as many as 4,000 women each year (14, 2). Twenty-five percent of the murders in this country are related to domestic violence (14). Domestic violence represents the largest segment of unreported crime in the United States (2). PMID- 11857883 TI - Reporting child maltreatment: dentistry's ethical responsibility. AB - The initial steps in the prevention of child maltreatment are early recognition and reporting. Reporting laws were not made to punish offenders, rather to help those children in need. Statistics indicate that among previously abused children who were returned home without intervention (reporting or counseling for the family), 5 percent were killed and 25 percent were seriously re-injured (22). While the nation's overall crime rate declined more than 21 percent from 1993 to 1997, the number of reports of child maltreatment increased by 8 percent with confirmed cases growing 4 percent during this same period (1). The dental profession must take a more active role on behalf of children in the prevention of this problem. When an incident arises that may result in a conflict between the dentist's personal interests and the well being of a patient, the dentist must always act in the best interest of the patient. Children have the right to grow up in a loving and nurturing environment. As dentists, we have the moral, ethical, and legally mandated responsibility to help protect this right through the recognition and reporting of child abuse and neglect. PMID- 11857885 TI - Elder mistreatment--what should dentists know? PMID- 11857886 TI - I don't understand it. PMID- 11857887 TI - Violence intervention programs: the Parkland Domestic Violence Project. PMID- 11857888 TI - Abuse. PMID- 11857889 TI - Dentistry's role in family violence prevention. PMID- 11857890 TI - Oral and maxillofacial pathology case of the month. Adenoid cystic carcinoma. PMID- 11857891 TI - A review of changes in the American Heart Association's "Guidelines 2000" for CPR (BLS). PMID- 11857892 TI - The spectrum of family violence. PMID- 11857893 TI - Long-term care planning: misconceptions and myths. PMID- 11857895 TI - Hears to your health. A sensor lets researchers listen for germs. PMID- 11857894 TI - Treat AIDS globally. PMID- 11857896 TI - Throw the switch? New vaccines may not be a reason to keep smallpox stocks. PMID- 11857897 TI - Defying gravity. A small Swiss firm develops an innovative G suit for fighter pilots. PMID- 11857898 TI - Who owns you? A mock trial explores the intersection of patents and genetic property rights. PMID- 11857899 TI - The worldwide computer. PMID- 11857901 TI - The cosmic reality check. PMID- 11857900 TI - Attacking anthrax. PMID- 11857903 TI - Repeated blows. PMID- 11857902 TI - Scars that won't heal: the neurobiology of child abuse. PMID- 11857904 TI - How should reading be taught? PMID- 11857905 TI - [Molecular structures and functional properties of cell-surface receptors]. PMID- 11857906 TI - [Signal transduction mechanisms of hormones through membrane receptors]. AB - Hormones exert their effect on cells either via membrane receptors or intracellular receptors. This paper aims to review membrane receptors and the intracellular signal transduction mechanisms. Membrane receptors could be classified according to their structural characteristics and the way they initiate the intracellular signal transduction. These include 1) Seven transmembrane(or G-protein coupled) receptors--heterotrimeric G-proteins- effector, system, 2) Receptor tyrosine kinases--protein-protein interaction through SH2, SH3, and PTB domain--MAP kinase cascades and PI3-kinase pathways, 3) Cytokine receptors--JAK--STAT pathways, 4) Receptors of the TGF- beta superfamily -SMAD pathways, 5) Apoptosis-related receptors--caspase pathways, and 6) ligand gated ion channels. There are growing knowledge of cross-talks between these pathways. It is being recognized that steroid hormones have distinct membrane receptors, which mediate rapid, nongenomic effect. PMID- 11857907 TI - [Molecular mechanism of hormone-dependent transcriptional regulation of nuclear receptor]. AB - Various actions of steroids, thyroid, vitamin D and retinoic acid are mediated by cognate nuclear hormone receptors. Hormone-liganded receptors bind to hormone response element in transcription regulatory region and homodimerize or heterodimerize with retinoid X receptor and regulate transcription of target genes. Conformational change of nuclear receptor after ligand-binding gives association with coactivators which possess histone acetyltransferase activity and are necessary for hormone-dependent transactivation. In the absence of hormone, corepressors, which associate with histone deacetyltransferase, interact to nuclear hormone receptors and repress the transcription. All these factors have important roles in the signaling of nuclear receptor and may be involved in endocrine dysfunction and disease. PMID- 11857908 TI - [Nuclear receptors in the pathophysiology of endocrine tumors]. AB - Nuclear orphan receptors COUP-TF and DAX-1 negatively regulate the transcriptional activity of SF-1, a steroidogenic cell-specific activator of various steroidogenic cytochrome P-450 genes. Expression of COUP-TF and DAX-1 are greatly decreased in cortisol-producing adrenocortical adenomas, while CYP17 is overexpressed. In contrast, DAX-1 expression is increased and SF-1 expression is decreased in deoxycorticosterone-producing adrenocortical adenomas, where CYP17 expression is low. These data suggest that DAX-1 and/or COUP-TF may repress CYP17 expression in adrenocortical tumors. In addition, SF-1 is expressed in the human pituitary gonadotroph and pituitary adenomas of gonadotrophic origin. SF-1 expression is associated with the in vitro secretion of LH in pituitary adenomas. Opposing effects of SF-1 and DAX-1 and/or COUP-TF may regulate the differential responses of various SF-1 target genes in different endocrine tumors. PMID- 11857909 TI - [Molecular defects in plasma membrane hormone receptors]. AB - The biological effects of hormones are mediated by plasma membrane receptors which transmit extracellular signals to the cytoplasm and nucleus. Mutations in plasma membrane receptors can affect normal signal transduction with loss-of function mutations leading to hormone resistance and gain-of-function mutations leading to constitutive activation of signaling pathways. The loss-of-function mutations leading to familial hormone resistance disorders are germline in origin whereas the gain-of-function mutations leading to constitutively active receptors are somatic. G-protein coupled receptors (GPCR) comprise a large superfamily of proteins characterized by seven transmembrane-spanning segments and interaction with GTP-binding(G) proteins. Mutations in GPCRs have been associated with dwarfism, congenital hyperthyroidism or hypothyroidism, nephrogenic diabetes insipidus, obesity, resistance to TSH, LH, FSH and ACTH, Jansen's metaphyseal and Blomstrand's chondrodysplasia, autosomal dominant hypoparathyroidism, and neonatal severe hyperparathyroidism. Mutations in other families of receptors which are characterized into one spanning-transmembrane receptor can result in resistance to insulin, GH, leptin and AMH. This review summarizes the molecular defects in plasma membrane hormone receptors in a large number of clinical disorders. PMID- 11857910 TI - [The etiology of steroid hormone insensitivity: nuclear cofactors and subnuclear architecture]. AB - Steroid hormone receptors interact with the basal transcription machinery(BTM) via intermediary cofactor complexes. Two complexes, a CBP/p300 complex with histone acetyltransferase(HAT) and a TRAP/DRIP/ARC/SMCC complex without HAT are identified, and a multistep model is proposed in which the two complexes interact sequentially with nuclear receptors. Recent advances in live-cell imaging techniques have revealed the interrelationship between subnuclear architecture and gene transcription, especially, nuclear receptors are known to form foci in transcriptionally activated state. These advances have developed a novel concept in a disorder of steroid hormone insensitivity. Disruption of the transmission from the AF-1 domain of the androgen receptor to BTM was revealed in a patient with androgen insensitivity who has no mutations in the androgen receptor gene. PMID- 11857911 TI - [Familial glucocorticoid deficiency due to the ACTH receptor gene mutations]. AB - Familial glucocorticoid deficiency(FGD) is a rare autosomal recessive disorder, characterized by resistance to ACTH leading to glucocorticoid deficiency, but not mineralocorticoid deficiency. Recently, mutations in the ACTH receptor gene were identified in several families with FGD. Thus far, twelve missense mutations, one nonsense mutation and three frameshift mutations causing FGD were described. Functional expression studies demonstrated that most of the missense mutations results in loss of specific binding to ACTH and impaired production of cAMP in response to ACTH. However, the genotype-phenotype correlation was poor. Interestingly and unexpectedly, the FGD patients with the ACTH receptor gene mutations were shown to be tall. PMID- 11857912 TI - [Inactivating and activating mutations of the human LH/hCG receptor leading to male pseudohermaphroditism and familial male-limited precocious puberty]. AB - The human luteinizing hormone/chorionic gonadotropin receptor(LHR) plays an important role in normal reproductive function in humans. Binding of LH to the LHR triggers production of intracellular c-AMP resulting in transcriptional activation of LH responsive genes. Mutations of the human LHR receptor lead to the development of three clinical conditions, namely, Leydig cell hypoplasia, hypergonadotropic hypogonadism and primary amenorrhea, and familial male-limited precocious puberty(FMPP). These mutations cause the receptor to lose its signal transduction activity in the first two conditions and to gain constitutive activity in the third condition. Leydig cell hypoplasia is an autosomal recessive disorder leading to male under-virilization. FMPP is an autosomal dominant disorder, characterized by excessive secretion of testosterone by Leydig cells and pubertal development at a very young age. Sporadic cases are also reported. PMID- 11857913 TI - [Mutations in the follicle-stimulating hormone receptor genes in patients with gonadal dysfunction]. AB - Follicle-stimulating hormone(FSH) plays important roles in gametogenesis and steroidogenesis in human gonads. Both activating and inactivating mutations have been detected only a few number in the gene for the FSH receptor. Inactivating mutations in the gene for the FSH receptor are involved in some female patients of hypergonadotropic hypogonadism with infertility. Only one activating mutation of FSH receptor was reported to have fertile functions in a hypophysectomized man. This article describes the reported genetic alterations of FSH receptor in humans and reviews how help us to understanding the molecular biology in the pathogenesis of gonadal dysfunction. PMID- 11857914 TI - [Congenital central hypothyroidism due to the mutations of the thyrotropin releasing hormone receptor gene]. AB - The TRH receptor gene is a single copy gene containing three exons and two introns and was assigned to the chromosome 8q23. A patient with congenital central hypothyroidism due to the mutations of the thyrotropin-releasing hormone receptor gene has been reported by Collu, et al: in 1997. His sole manifestation was short stature, and his plasma TSH and prolactin levels did not increase in TRH test. He was found to be a compound heterozygote of the mutations of the TRH receptor gene. Expression study using COS1 cells and HEK293 cells revealed that both mutations reduced or abolished the receptor activity. PMID- 11857915 TI - [Familial congenital hypothyroidism due to loss of function mutation of the thyrotropin receptor (resistance to thyrotropin)]. AB - The TSH receptor belongs to a subfamily of G protein-coupled receptors that mediates the effect of TSH in thyroid development and hormone synthesis. Similarly to many other G protein--coupled receptors, both gain and loss of function mutations have been found in the TSH receptor. Loss of function mutations in the TSH receptor gene were first described in three siblings with resistance to TSH in 1995. To date, 13 pedigrees with TSH receptor mutations of loss of function type have been reported. The phenotype of the patients varies from euthyroid hyperthyrotropinemia to severe congenital hypothyroidism with thyroid hypoplasia. PMID- 11857916 TI - [Non-autoimmune hyperthyroidism and hyperfunctioning thyroid adenomas caused by activating mutation of the thyrotropin receptor]. AB - Constitutively activating mutations in the TSH receptor gene have been found in various disorders. Somatic mutations were identified in autonomously functioning thyroid nodules(AFTNs) and toxic multinodular goiters(TMNGs). Germline mutations were identified in autosomal dominantly inherited non-autoimmune hyperthyroidism and sporadic congenital hyperthyroidism. Most of activating TSH receptor mutations were identified in western countries. Only one Japanese family has been reported to have an activating TSH receptor mutation. Previous very poor and insensitive study identified no somatic TSH receptor activating mutations in 45 AFTNs and TMNGs developed in Japanese. Our recent study completely reversed their observation; about half of AFTNs in Japanese had activating mutations. PMID- 11857917 TI - [Isolated GH deficiency due to inactivating mutation of GHRH receptor]. AB - Pituitary GH secretion is controlled by hypothalamic GHRH via its own receptor. Recently, inactivating mutants of GHRH receptor were found in subjects with isolated GH deficiency. These subjects show less typical facial appearance than those with GH gene deletion or GH resistance. They respond well to GH treatment. We found a novel mutation of GHRH receptor in a Japanese boy with IGHD. The boy was with four base pair deletion in exon 12 of GHRH receptor that caused frame shift and premature stop codon resulted in a C-terminally truncated receptor formation. This novel mutation indicated the critical role of intracellular C terminus of the GHRH receptor. PMID- 11857919 TI - [Truncated growth hormone receptor mutations function as dominant-negative inhibitors of the full-length receptor and cause genetic short stature]. AB - Truncated growth hormone receptor (GHR) mutations that lack the majority of the cytoplasmic domain have been identified in familial short stature and same truncated GHR isoforms generated by alternative splicing in a number of normal human tissues. The binding affinity of the truncated GHR was similar to that seen for the full-length receptor and the truncated GHR showed a much greater expression on the cell surface, which is consistent with lack of internalization. These truncated receptors function as dominant-negative inhibitors of the growth hormone (GH) signal and generate large amounts of GH binding protein (GHBP). Thus, truncated GHR isoform may be important as a physiological regulator of GH signaling in addition to providing a mechanism for the production of GHBP. PMID- 11857918 TI - [Laron syndrome by loss of functions of growth hormone receptor]. AB - Laron syndrome, characterized by a short stature with raised serum levels of growth hormone but extremely low levels of insulin-like growth factor I, is caused by genetic defects of growth hormone receptor, including loss of functions of growth hormone receptor and abnormal growth hormone receptor acting in a dominant negative manner. This chapter focused on loss of functions of growth hormone receptor causing disruption of growth hormone binding and intracellular signalling. Most mutations and deletions have been found in extracellular domains of growth hormone receptor. Serum levels of growth hormone binding protein, cleaved from the extracellular portion of the growth hormone receptor, are typically decreased in most patients but are normal or high in some patients. PMID- 11857920 TI - [Normosmic idiopathic hypogonadotropic hypogonadism with GnRH receptor mutation(review)]. AB - Mutations in gonadotropin releasing hormone receptor(GnRHR) gene(chromosome location, 4q21.2) cause autosomal recessive idiopathic hypogonadotropic hypogonadism (IHH) without anosmia. The spectrum of phenotypes is much broader, the mechanism of which still remains clarified yet. The prevalence may be 2.2% in all families with IHH, but 7.1% in families in which the affected female is present. The frequency may be common in familial case(40%) and infrequent in general sporadic cases(16.7%). Functional mutations of the GnRHR are distributed widely throughout the protein, although the hot spots are Gln(106) Arg and Arg (262) Gln. The same mutations cause different gonadotropin secretion capability, in which hyperstimulated pulsatile GnRH treatment could bring ovulation and pregnancy in some cases. PMID- 11857921 TI - [Familial hypocalciuric hypercalcemia and neonatal severe hyperparathyroidism caused by inactivating mutations of calcium-sensing receptor]. AB - Calcium-sensing receptor (CaSR) plays an essential role in regulating secretion of parathyroid hormone. After the identification of CaSR, some cases of familial hypocalciuric hypercalcemia (FHH) were shown to have heterozygous inactivating mutations of CaSR. However, linkage study showed that there are additional two genetic loci for FHH in addition to the chromosomal location of CaSR gene. Furthermore, one family with heterozygous inactivating mutation of CaSR was shown to exhibit hypercalcemia with hypercalciuria. Therefore, heterozygous inactivating mutation of CaSR is not synonymous with FHH. In addition, patients with neonatal severe hyperparathyroidism were shown to have homozygous or compound heterozygous inactivating mutations of CaSR. PMID- 11857922 TI - [Familial hypoparathyroidism due to activating mutations in the calcium-sensing receptor gene]. AB - The cloning of the extracellular calcium-sensing receptor(CaSR) has helped to define a key component in the control of the calcium homeostasis. Gain-of function mutations in the CaSR gene were identified as the cause of autosomal dominant hypocalcemia (ADH). This clinical condition is compatible with hypoparathyroidism because of low PTH levels compared with serum calcium levels. Until now, 21 missense mutations of the CaSR were identified as a cause of hypoparathyroidism (HP). Mutations present in transmembrane domain may produce more severe hypocalcemia than those present in other domains. We emphasize that it is important to differentiate ADH from PTH-deficient HP, because treatment with vitamin D to correct the hypocalcemia in the former may lead to more severe hypercalciuria, nephrocalcinosis, and renal impairment. PMID- 11857923 TI - [PTH/PTHrP receptor and pseudohypoparathyroidism]. AB - The PTH/PTHrP receptor, a G protein coupled receptor, mediates the actions of PTH and PTHrP. Homozygous inactivating mutations in this receptor cause a rare disorder called Blomstrand chondrodystrophy. Since this disease is lethal in utero or perinatal period, the existence of PTH resistance has not been verified. Inactivating mutations in PTH/PTHrP receptor have not been found in the patients with pseudohypoparathyroidism (PsH), a group of disorders characterized by biochemical hypoparathyroidism and unresponsiveness to PTH. In patients with PsH type Ia, numerous heterozygous inactivating mutations have been identified in the gene (GNAS1) encoding the alpha-subunit of Gs protein (Gs alpha) which couples multiple receptors to the stimulation of adenylate cyclase. It is suggested that some abnormality in GNAS1 outside the coding exons may lead to PsH type Ib. To explain the heredity and phenotypic variation of PsH, tissue- or cell-specific imprinting of Gs alpha has been implicated. PMID- 11857924 TI - [Leprechaunism caused by mutations in the insulin receptor gene]. AB - Leprechaunism represents the most severe form of insulin resistance syndrome, manifesting abnormal glucose metabolism and intrauterine, postnatal growth retardation. Mutations in both alleles of the insulin receptor gene have been identified. Recombinant human IGF-I treatment could prevent postnatal growth retardation and normalize glucose metabolism, however there are few reports of long-term treatment with IGF-I. We have a case of compound heterozygous mutations of the insulin receptor gene, who has been treated with IGF-I more than 11 years. Relatively higher dose of IGF-I is necessary for maintaining sufficient serum IGF I levels. After 10 years' treatment with IGF-I, polycystic ovary, kidney enlargement, albuminuria and retinopathy are complicated in this patient. In this review, we summarized basic actions of insulin and insulin receptor, classification of mutations in the insulin receptor gene, clinical feature and our results of long-term treatment with IGF-I in leprechaunism. PMID- 11857925 TI - [Nephrogenic diabetes insipidus associated with mutations of vasopressin V2 receptors and aquaporin-2]. AB - Nephrogenic diabetes insipidus has two types of X-linked and autosomal recessive inheritance. The former is the mutations of arginine vasopressin (AVP) V2 receptors that have had 155 mutations in 239 families in the literature. The latter is the mutations of aquaporin-2(AQP-2) water channel, which have had 11 mutations. The functional analysis of V2 receptor mutations has resulted in two abnormalities. The mutated receptors retain in cytoplasma and can not fold into plasma membrane in most of AVP V2 receptor mutations. The other is that the mutated receptors, localized in plasma membrane, can not either bind to its ligand AVP or transduce its signal to the post-receptor pathway. Also, the mutated AQP-2 is functionally divided into two types of abnormality. In 10 out of 11 mutations, the mutated AQP-2 is located in endoplasmic reticulum or Golgi apparatus, and can not be translocated into apical plasma membrane. The mutated AQP-2 should functionally produce water permeability, if it could be routed into plasma membrane. Only one mutation of AQP-2 (T125M and G175R) can be folded in apical membrane, but it does not produce water permeability. Recently, the experimental trials have been begun for rescuing the mutated AVP V2 receptors or AQP-2. PMID- 11857926 TI - [Disorders caused by abnormalities of glucocorticoid receptors]. AB - Abnormalities of glucocorticoid receptors were assumed to cause resistance or hypersensitivity to glucocorticoid, while only glucocorticoid resistance has been described clinically. Patients with familial cortisol resistance have continuously elevated serum cortisol without any clinical manifestations of Cushing's syndrome due to hyposensitivity to cortisol in all tissues including the hypothalamus and the pituitary. Clinical symptoms of the disease are characterized by hypertension with hypokalemia and hyporeninemia, virilism in women, isosexual precocity in a boy and mild general fatigue. As the cause of the disease, a defect in glucocorticoid receptor affinity or binding capacity due to mutations in the glucocorticoid receptor gene has been reported. Another cause of the disease is the presence of heat labile glucocorticoid receptor. In 5 of 6 families with cortisol resistance reported so far, mutations of the glucocorticoid receptor gene have been demonstrated. PMID- 11857927 TI - [Genetic disorders caused by gain or loss of function of the mineralocorticoid receptor]. AB - Aldosterone has crucial role for sodium conservation in the kidney, salivary glands, sweat glands and colon. It exerts its effects via the mineralocorticoid receptor (MR) which belongs to the member of the nuclear receptor superfamily. Recently, genetic disorders have been reported to be caused by gain or loss of function of the MR. The amino acid substitution of the ligand-binding domain(S810L) of the MR resulted in the early-onset hypertension exacerbated by pregnancy. This mutation results in constitutive MR activity and alters receptor specificity for progesterone. Pseudohypoaldosteronism type 1 (PHA1) is characterized by congenital aldosterone resistance of the kidney and/or other mineralocorticoid target tissues, resulting in excessive salt wasting. The heterozygous nonsense or missense mutations were identified in the patients with autosomal dominant PHA1 and a sporadic PHA1. This suggests that the full expression of the MR is necessary for salt conservation. PMID- 11857928 TI - [Male pseudohermaphroditism due to disorder of androgen receptor function]. AB - Disorder of androgen receptor function, generally due to mutations in androgen receptor gene, is one of the causes of male pseudohermaphroditism. The disorder results in a wide spectrum of abnormalities of male sexual development ranging from phenotypic female to male with infertility depending on residual androgen receptor function. Clinical features, endocrinological data and treatment of patients with the disorder were discussed. PMID- 11857929 TI - [Pathophysiology of disease caused by mutation in estrogen receptors]. AB - Study of the mutation in estrogen receptor alpha gene in a man has been reported. This observation provides the new insights in the critical role of estrogen in the male for skeletal maturation and maintenance of bone mass. This new concept is supported by the findings on the patients with aromatase deficiency. In addition, I would like to stress the district differences in the clinical profiles between estrogen receptor alpha-deficient male and estrogen receptor alpha-knockout mouse. PMID- 11857930 TI - [Thyroid hormone receptors and its disorders]. AB - Resistance to thyroid hormone (RTH) is a genetic disease caused by mutations of thyroid hormone receptor beta gene. RTH is manifested by inappropriately elevated levels of serum thyrotropin in the presence of elevated circulating thyroid hormones. Most of the patients with RTH do not manifest remarkable clinical features, but may represent partial hypothyroidism. Sometimes even tachycardia, sweating, body weight loss or attention-deficit hyperactivity disorder are found. TR beta mutants, which have reduced T3-binding activities and aberrant interactions with transcriptional cofactors, act in a dominant negative fashion against wild type TRs. Although no human disease has been found to be caused by TR alpha gene mutations, TR alpha knockout mice suggest that TR alpha plays a major role in the regulation of heart rate and basal metabolism and TR alpha gene deficiency does not express the RTH phenotype. PMID- 11857931 TI - [Bone disease with vitamin D receptor abnormality]. AB - In humans, the vitamin D receptor (VDR) gene has been localized to the chromosomal locus 12q13-14. The gene is composed of a minimum of nine exons. Hereditary 1,25-dihydroxyvitamin D resistant rickets (HVDRR) known as vitamin D dependent rickets type II is a rare autosomal recessive disease that arises as a result of mutations in the gene encoding the VDR. Genetic factors play a key role in determining bone mass, which is an important predictor of osteoporosis. Recently, polymorphism at the VDR locus has been implicated as a genetic marker for bone mineral density. Vitamin D receptor gene start codon polymorphisms, and 3'-end region polymorphisms may modulate bone density. PMID- 11857932 TI - [DAX-1 abnormality]. AB - DAX-1 is an orphan nuclear receptor that plays a key role in the development and function of the adrenal gland and hypothalamic-pituitary-gonadal axis. Absence of DAX-1 results in X-linked adrenal hypoplasia congenita, a human inherited disorder characterized by adrenal insufficiency and hypogonadotropic hypogonadism. The DAX-1 gene may be responsible for a male-to-female sex reversal syndrome, referred to as dosage sensitive sex reversal(DSS), due to the duplication of a small region of human chromosome Xp21. Dax-1 and Sry have been shown to act antagonistically in the mouse system, where over-expression of Dax-1 leads to female development and increasing activity of Sry to male development. Although these data strongly implicate Dax-1 in sex determination, there is no evidence that DAX-1 is equivalent to DSS in human. PMID- 11857933 TI - [Endocrine disruptors and nuclear receptors]. AB - Endocrine dirsuptors are supposed to modulate function of nuclear receptors by binding their ligand-binding domain. As nuclear receptors are ligand dependent transcription factor, endocrine disruptors function as agonist or antagonist. Although actual mechanism of endocrine disruption is not well understood, several nuclear receptors are reported as targets of endocrine disruptors. In this review, the functions of nuclear receptors suspected as targets of endocrine disruptors are described. PMID- 11857934 TI - [Obesity induced by abnormality in leptin receptor and melanocortin-4 receptor]. AB - Obesity is a multifactorial disease that arises from complex interactions between genetic predisposition and environmental factors. It increases a risk of cardiovascular and metabolic diseases such as diabetes, hypertension, and hyperlipidemia. Recent molecular genetic studies have disclosed some monogenic forms of obesity in humans. Leptin directly exerts its anorexigenic effects on hypothalamic arcuate nucleus. alpha-melanocyte stimulating hormone (alpha-MSH) derived from proopiomelanocortin (POMC) and melanocortin-4 receptor (MC4-R) have been reported to be involved in the downstream of leptin actions. In this paper, we summarize the clinical characteristics and the mechanisms of obesity caused by genetic abnormalities in leptin receptor and melanocortin-4 receptor. PMID- 11857935 TI - [Neural transplantation for cerebral ischemia/infarction--the present situation and prospects]. AB - Cerebral infarction results in multiple symptoms including hemiplegia and cognitive disturbances. The central nervous system has a limited capacity for self-repair, thus there is a great interest in the possibility of repairing the central nervous system by neural transplantation. Two different types of cerebral infarction model are well investigated for neural transplantation. Several kinds of donor cells have been used to try to restore the brain damage after ischemic insult. Reconstruction of neural circuits by transplantation is an ideal goal for the treatment of cerebral infarction, but trophic action of transplantation is also expected. Amelioration of ischemia-induced brain damage and recovery of neural dysfunction are documented. However, there are several factors and problems to be solved for clinical application. PMID- 11857936 TI - [Neurosurgery in the 21st century--recent progress and perspectives in neurovascular intervention]. PMID- 11857937 TI - [Neurosurgery in the 21st century--pediatric neurosurgery; present and future]. PMID- 11857938 TI - [Spinal arteriovenous malformation]. PMID- 11857939 TI - [Monobloc fronto-facial advancement for craniosynostosis syndrome: discussion and viewpoint of a plastic surgeon using an internal distraction device]. PMID- 11857940 TI - [Cranioplasty using the patient's autogenous bone preserved by freezing--an examination of post-operative infection rates]. AB - The current technique for cranioplasty using artificial bone requires further improvement with regard to infection, strength and comfort through good fitting. We have carried out cranioplasty using the patient's autogenous bone flap obtained during first surgery. It was immersed in 200 mg of Amikacin Sulphate, and frozen at -16 degrees C until its use in cranioplasty. From 1980 to 1998, cranioplasty has been carried out on 206 patients. They consisted of 118 males and 88 females, and their age ranged in our institute from 1 to 81; average age 51.1. Ruptured aneurysm (48%), head injury (14%), intracranial hemorrhage (23%) and cerebral infarction (12%) were the major causes requiring decompression surgery. We analyse the bone preservation period and the time between cranioplasty and the onset of infection. The infection rates per bone preservation periods, the causes of decompression and age groups are studied. Of the 208 case studies, infection necessitating bone removal or debridement was noted in 8 cases (3.88%). Average bone preservation period in the infected group was 31.1 days as compared with 54.9 days for the non-infected group (p < 0.05). Not patient age but the type of head injury is also a significant factor in post cranioplasty infection. PMID- 11857941 TI - [Preemptive analgesia for postoperative pain after frontotemporal craniotomy]. AB - Two thirds of patients suffer from moderate to severe pain after frontotemporal craniotomy. We think neurosurgeons must try to reduce the postoperative pain, which may induce postoperative hypertension, restlessness, and pathological pains. To investigate how preemptive analgesia effects postoperative pain, we assessed the pain in 20 consecutive patients who underwent neck clipping for non ruptured cerebral aneurysms of anterior circulation systems by frontotemporal craniotomies. Ten patients underwent preemptive analgesia with four procedures (preemptive group) as follows, 1) oral administration of long-acting non-steroid anti inflammatory drug (NSAID, ampiroxicam) two hours before the surgical operation, 2) nerve blockades of the supra-orbital nerve and the infra-orbital nerve by bupivacaine, 3) local anesthesia of the scalp along the marker of a skin incision by xylocaine, 4) local anesthesia by bupivacaine along a skin incision after the skin closure. Ten patients of the control group underwent only procedure No. 3. Visual analog pain score (VAS) for postoperative pain 6, 12, and 24 hours, and 3, 5, 7, and 14 days after operation and NSAID administration for the pain were evaluated. Patients of the preemptive group had significantly less postoperative pain during the whole post-surgery period and required less administration of NSAID than the control group. Preemptive analgesia procedures No. 1, 2 and 4 reduced the postoperative pain and the total administration of NSAID. Postoperative pain may be reduced after other types of brain surgery, with proper nerve blocks like procedure No. 2, procedures No. 1, 3 and 4. PMID- 11857942 TI - [A ruptured distal aneurysm, thought to be a mycotic aneurysm, associated with acute subdural hematoma: case report and review of the literature]. AB - A case is described of a ruptured intracranial mycotic aneurysm in the distal middle cerebral artery associated with an acute subdural hematoma. A 55-year-old woman, with a history of tuberculous meningitis at the age of 7, presented left hemiparesis. She was in a state of semi-coma. Computed tomography scan showed a right acute subdural hematoma. Magnetic resonance angiography revealed no cerebral aneurysm. Emergency external decompression was performed and a distal central artery aneurysm was found and was resected during the operation. Numerous areas of hemosiderin pigmentation were recognized around the aneurysm and in the subarachnoid space. Microscopic examination of the aneurysmal sections showed neither elastic lamina nor membranous structures and no infiltration of any inflammatory cells. We think this aneurysm is a pseudoaneurysm. The incidence of ruptured mycotic aneurysms presenting with acute subdural hematoma is extremely low. There have been only four previously reported cases as far as I can ascertain. This case and a review of the literature are discussed. PMID- 11857943 TI - [A case report of transverso-arthropediculectomy for thoracic spondylosis]. AB - We report a case of a 23-year-old male with thoracic spondylosis at the T2/3 level. He complained of lumbago and gait disturbance (spastic paraparesis and sensory disturbance below the T4 level). Preoperative CT myelography showed a posterolateral spur at the left paramedian site which was compressing the spinal cord. We performed drilling of the spur using the posterolateral approach, and decompressed the spinal cord. After surgery, the motor weakness and sensory disturbance gradually improved and spasticity was reduced. Postoperative CT showed complete decompression of the spinal cord and no spur. In cases of upper thoracic lesions, there are various kinds of posterolateral approaches for decompression of the ventral side of the spinal cord. We adopted transverso arthropediculectomy for this case, because this approach is minimally invasive of the costotransverse joint and the spinal cord. We discuss the other posterolateral approaches, in terms of merits and demerits for the vertebra and spinal cord. PMID- 11857944 TI - [Intramedullary spinal cord metastasis: two case reports and literature review]. AB - Intramedullary spinal cord metastasis was thought to be rare, but the advent of MR imaging and treatment for malignancy have revealed that it is more frequent than we previously thought. We report two cases of intramedullary spinal cord metastasis from lung and rectal colon carcinoma. Their symptoms were aggravated rapidly. We performed surgical treatment in both cases; one was grosstotal and the other was subtotal resection. After the operation, symptoms improved mildly in one patient and worsened in the other patient. Both died 3 months after surgery, because of respiratory disturbance due to brain stem compression and lung metastasis. In conclusion, MRI is highly useful for making a diagnosis of intramedullary spinal metastatic tumors. However, it is still difficult to treat such a patient by surgery alone. Further postoperative adjuvant therapy will be necessary to improve the prognosis of such patients. PMID- 11857945 TI - [A case of Rathke's cleft cyst in association with a ruptured aneurysm of the anterior cerebral artery mimicking pituitary apoplexy]. AB - We present a rare case of a Rathke's cleft cyst in association with a ruptured aneurysm of the anterior cerebral artery (ACA). A 44-year-old man suffered from sudden onset of headache. Initial computed tomographic (CT) scan revealed a high density mass lesion in the suprasellar region and a diffuse high-density area in the basal cistern. Using emergent magnetic resonance imaging (MRI), we found a cyst showed homogeneously high and iso-intensity on T1 and T2-weighted image, respectively. The cyst showed no enhancement of the cyst wall, but on Gd-DTPA, it was shown to compress the normal pituitary gland. Angiography showed an aneurysm at the A1 portion of the left ACA. Based on/these findings, we were able to diagnose Rathke's cleft cyst and a ruptured aneurysm. An operation was performed through the interhemispheric approach. The suprasellar cystic mass compressed the optic nerves and chiasm upward. Neck clipping of the aneurysm and opening of the cyst were performed. We confirmed the cause of the subarachnoid hemorrhage as being a ruptured aneurysm at the A1 portion of the left ACA. Histological diagnosis was Rathke's cleft cyst. Postoperative course was uneventful. There has been only one reported case of Rathke's cleft cyst in association with a ruptured aneurysm. When we encounter a case presenting subarachnoid hemorrhage with suprasellar mass and intracerebral aneurysm, we must discriminate between ruptured aneurysm and pituitary apoplexy in the acute stage as the cause of the subarachnoid hemorrhage. If the mass is Rathke's cleft cyst, we speculate that the cause of the subarachnoid hemorrhage is a ruptured aneurysm, because there are no reports of Rathke's cleft cyst with subarachnoid hemorrhage. PMID- 11857946 TI - [Traumatic vertebral arteriovenous fistula caused by an injury due to penetration by pieces of glass: case report]. AB - We reported a case of a 79-year-old male with traumatic vertebral arteriovenous fistula. The patient was admitted with hemorrhagic shock due to arterial bleeding caused by a penetrating injury of the neck. Cervical CT showed free air at the level of the lamina of C1. Injury of the vertebral artery was suspected. The right vertebral angiogram showed an arteriovenous fistula between the V3 portion of the right vertebral artery and the suboccipital venous plexus with retrograde intracranial venous drainage. The patient underwent endovascular treatment of the parent vertebral artery occlusion, which successfully obliterated the arteriovenous fistula. PMID- 11857947 TI - [A case report of a scalp dermoid cyst containing watery-clear fluid]. AB - We report a case of a 7-month-old female with a dermoid cyst on the anterior fontanelle. She was born with a vacuum extractor. Two weeks later, her scalp on the anterior fontanelle bulged. A doctor was consulted when she was 3 months old, because the tumor was enlarging. Magnetic resonance image (MRI) showed a cystic mass on the anterior fontanelle. She was brought to our institute. At the first medical examination, she was alert and had no neurological deficit. The tumor was suspected to be a sinus pericranii or a pseudo meningocele. She was observed as an out patient, but the tumoral size become more enlarged. When she was seven months old, we punctured the cystic tumor and the tumor collapsed. But, two weeks later, it enlarged again. The cystic fluid was watery clear and the composition differed from cerebrospinal fluid (CSF). The tumor was resected totally. The histological examination revealed a dermoid cyst which involved ducts of the eccrine glands and folliculi pili. The cystic fluid was thought to be secreted from the eccrine glands. PMID- 11857948 TI - [A case of traumatic sinus pericranii]. AB - Sinus pericranii is a rare vascular anomaly involving an abnormal communication between the extracranial and intracranial circulations. A 33-year-old woman presented with a soft tissue mass at the left frontal region. It was associated with head trauma when she was 12-year-old. The mass had gradually grown and become painful for 20 years. She underwent surgical resection of the mass successfully. This report discusses traumatic sinus pericranii. PMID- 11857949 TI - [Outcome of hepatic nodules less than 10 mm in size detected by ultrasonography in patients with chronic liver disease]. AB - The number of hepatic nodules detected by ultrasonography in patients with chronic liver disease has increased. Although it potentially is important to understand the outcome of small solitary hepatic nodules, they have not been fully investigated. In this study, we evaluated retrospectively the outcome of 35 solitary hepatic nodules smaller than 10 mm in 35 patients with chronic liver disease. The follow-up period ranged from 24 to 94 months, and, during follow-up, 8 of 35 nodules (23%) increased in size, 17 nodules (49%) showed no change, and 10 nodules (29%) disappeared. It took less than 12 months for 5 nodules to increase in size, and less than 24 months for 3 nodules. There was no significant difference between the ultrasonographic findings (echogenicity and size) of the nodules and their outcome. We conclude that follow-up observation of at least 24 months is essential to evaluate the outcome of solitary hepatic nodules. PMID- 11857950 TI - [Japanese radiological report creation with continuous speech recognition]. AB - Ten Japanese radiological reports consisting of 1381 characters (681 words) were created by two board-certified radiologists who used conventional typing and a continuous speech-recognition system called AmiVoice (Advanced Media, Inc., Tokyo, Japan). The two radiologists had not had any special training prior to their use of the continuous speech-recognition system. The model of speech-to text analysis was generated from 22,589 radiological reports (5.7 MB). Dedicated pronunciations for loan words (i.e., English words) were registered by a board certified radiologist in consideration of variations in Japanese pronunciation. Misrecognition occurred in 40 of 1362 words, corresponding to a 97.1% rate of accuracy of recognition. The average speech recognition time per report was 31.3 sec, and the additional time required for corrections was 25.0 sec. The total speech input time of 56.2 sec was much less than the conventional input time of 142.8 sec for typing. Continuous speech recognition is faster than typing, even considering the additional time required for corrections, and is acceptable in view of the overall reduction in report turn-around time. PMID- 11857951 TI - [Second malignancies following radiotherapy: an analysis of 54 cases accumulated by mail survey in Japan]. AB - The increasing incidence of second malignant neoplasms after radiotherapy, while due in part to increasing numbers of survivors, is also thought to be related to new modalities of radiotherapy and/or increasingly more intensive combined modality treatment. From a mail survey conducted in 2000 concerning secondary neoplasms following radiotherapy, 62 patients were collected from 22 hospitals in Japan. The following patients were excluded: benign (4 cases) or unknown (2) first primary diseases, unknown histology of a second malignancy (1), and short latent period (from initial radiotherapy to diagnosis of second neoplasm) (1). Fifty-four patients with second malignancies were analyzed. The most common histology of second malignancies was squamous cell carcinoma (24 cases), followed by sarcoma (16), glioma (5), adenocarcinoma (3), leukemia (3), and others (3). The mean latent period was 17.7 (2-38) years. We investigated the correlation of the latent period with patient's characteristics or initial therapeutic factors. Multivariate analysis revealed that the latent period was significantly shortened in patients with combined chemotherapy and radiotherapy. PMID- 11857952 TI - [Visionary choice of Japanese radiologists and radiological technologists in 21st century]. AB - Since X-ray CT appeared in radiology practice in the 1970s, there has been rapid progress in imaging modalities such as US, MRI, PET, and PACS. With these new modalities, radiology has strengthened its advantage in medical practice. In earlier times, it was said "ohne Roentgen, keine Medizin". This is still true today, and undoubtedly will be true even in the future. Medical practice can no longer be carried out in the absence of the new imaging technologies. On the other hand, sophisticated computer technology has engendered a new fight for turf among the medical specialties. To win this battle, I consider improved mutual understanding, close cooperation, and esprit de corps between radiologists and radiological technologists to be of utmost importance. This article describes the following: (1) what a medical team should be, (2) how a good medical team should be organized, (3) how important a four-year course in medical technology is, (4) how our survival can be ensured in the new turf battle with advanced medical technology, (5) how JMCP should be run in the future, (6) how important it is for Japanese radiology to show a strong presence in the Asian-Oceanian Society of Radiology, (7) how we can survive in a digital world, and (8) how we should relate to the recent social environment surrounding us. PMID- 11857953 TI - [Selective intraarterial infusion chemotherapy using a combined MRI-angiography system for head and neck cancers]. AB - We performed selective intraarterial infusion chemotherapy using a combined MRI angiography system for head and neck cancers. In three patients, three or five infusions of CDDP (100 mg/body) were administered to the feeding arteries selectively. For the evaluation of drug distribution, MRI during arteriography through the infusion pump was performed before CDDP administration. When a distribution mismatch was found, arterial selection was attempted again under a mechanically unstable C-arm system, and further evaluation under an MR system was carried out. Thus, more ideal treatment could be provided. We consider MRI during arteriography to be useful in assessing for distribution during intraarterial chemotherapy. PMID- 11857954 TI - [MR imaging in obstetrics and gynecology]. AB - Excellent tissue contrast in MRI permits noninvasive visualization of the normal anatomy, physiology, and disease processes of the female pelvis. Although MRI has various disadvantages, including its higher cost compared with sonography and CT and its long scanning time, many studies have reported a positive impact of MRI on treatment decisions and net cost analysis in the evaluation of gynecologic diseases. In addition to traditional static images, recent advances in ultrafast MR imaging have provided MR angiography, MR urography, and kinematic images of the pelvis. Kinematic evaluation of the pelvis by MR imaging is an epoch-making event. Kinematic images can be obtained by a cine-mode display of many images obtained by ultrafast imaging techniques such as FISP, HASTE, SSFSE, and others. Pelvic floor descent, an adhesion caused by endometriosis or surgery, and uterine peristalsis can be clearly shown on kinematic images. Since disorders in uterine peristalsis are hypothesized to be the causes of a variety of clinical symptoms such as fertility problems and dysmenorrhea, ultrafast MRI may be a powerful tool for elucidating the functions of the uterus in the future. PMID- 11857955 TI - [Creation of functional organic compounds and their applications]. AB - Our studies on creation of functional organic compounds and their applications, have focused on three areas, namely, (A) organic chemical studies on VD (vitamin D) analogues, (B) studies on solitary wasp venoms, and (C) studies on functional building blocks for organic synthesis. In the first area, several novel and important vitamin D analogues were synthesized and biologically evaluated, and their high VDR (vitamin D receptor) binding affinities were discussed on the basis of conformational analysis and docking study by Molecular Mechanics Calculation to the LBD (ligand binding domain) of VDR: These compounds include 24,24-difluoro-1 alpha,25-dihydroxy-VD3 (2) (an antimetabolism agent, the first VD analogue having higher potency than the natural hormone (1)), 2 alpha-methyl-1 alpha,25-dihydroxy-VD3 (42b) (the first A-ring-modified VD analogue exhibiting stronger VDR affinity than 1) and its 20-epimer (43b) (a VD analogue having a highest HL-60 cell differentiation inducing activity with a relatively low calcemic effect), and 2 alpha-(omega-hydroxypropyl)-1 alpha,25-dihydroxy-VD3 (exceptionally high calcemic effect). In the second area, we isolated and determined the structure of pompilidotoxins (76, 77), novel peptide neurotoxins in solitary wasp venoms. In the third area, we created furan-fused 3-sulfolene, 4H, 6H-thieno [3,4-c]-furan 5,5-dioxide and pyrrole-fused 3-sulfolene (96), 3,5 dihydro-1H-thieno[3,4-c] pyrrole 2,2-dioxide (125), and studied their inter- and intramolecular Diels-Alder reactions. PMID- 11857956 TI - [Preparation and evaluation of press-coated aminophylline tablet using crystalline cellulose and polyethylene glycol in the outer shell for timed release dosage forms]. AB - In an attempt to achieve chronopharmacotherapy for asthma, press-coated tablets (250 mg), which contained aminophylline in the core tablet in the form of low substituted hydroxypropylcellulose (L-HPC) and coated with crystalline cellulose (PH-102) and polyethylene glycol (PEG) at various molecular weights and mixing ratios in the amounts of PH-102 and PEG as the outer shell (press-coating material), were prepared (chronopharmaceutics). Their applicability as timed release (delayed-release) tablets with a lag time of disintegration and a subsequent rapid drug release phase was investigated. Various types of press coated tablets were prepared using a tableting machine, and their aminophylline dissolution profiles were evaluated by the JP paddle method. Tablets with the timed-release characteristics could be prepared, and the lag time of disintegration was prolonged as the molecular weight and the amount of PEG, for example PEG 500,000, in the outer shell were increased. The lag time of disintegration could be controlled by the above-mentioned method, however, the pH of the medium had no effect on disintegration of the tablet and dissolution behavior of theophylline. The press-coated tablet (core tablet:aminophylline 50 mg, L-HPC and PEG 6000; outer shell:PH-102:PEG = 8:2 200 mg) with the timed release characteristics was administered orally to rabbits for an in vivo test. Theophylline was first detected in plasma more than 2 h after administration; thus, this tablet showed a timed-release characteristics in the gastrointestinal tract. The time (tmax) required to reach the maximum plasma theophylline concentration (Cmax) observed after administration of the press-coated tablet was significantly (p < 0.05) delayed compared with that observed after administration of aminophylline solution in the control experiment. However, there was no difference in Cmax and area under the plasma theophylline concentration-time curve (AUC0-->24) between the press-coated tablet and aminophylline solution. These results suggest that the press-coated aminophylline tablet (with the timed release characteristic) offers a promising forms of theophylline chronotherapy for asthma. PMID- 11857957 TI - [Effects of Byakko-ka-ninjin-to, Bofu-tsusho-san and Gorei-san on blood glucose level, water intake and urine volume in KKAy mice]. AB - KKAy mouse, a genetic animal model of diabetes mellitus, is characterized by not only hyperglycemia, but also polydipsia and polyuria like human diabetes mellitus. In this study, we investigated the effects of Byakko-ka-ninjin-to (BKN), Bofu-tsusho-san (BOF) and Gorei-san (GRE) on blood glucose level, water intake and urine volume in KKAy mice. All these Kampo medicines were administered to the mice as the food admixture containing 1.5% or 4.5% of weight of food for 4 weeks. BKN at 1.5% and 4.5% decreased water intake without affecting blood glucose level. BOF at 1.5% showed the same effects as BKN did, but it decreased both water intake and blood glucose level at 4.5%. Furthermore, BOF at 4.5% obviously, but BKN at 4.5% slightly, decreased urine volume. However, GRE at 4.5% did not show any effects on blood glucose level, water intake and urine volume. From these results, BKN is confirmed to be effective against the polydipsia accompanied by diabetes mellitus, while BOF is suggested to be effective against not only polydipsia, but also polyuria and hyperglycemia in diabetes mellitus. PMID- 11857958 TI - [Studies on the risk factors for fenofibrate-induced elevation of liver function tests]. AB - We evaluated risk factors for elevation of liver function test values after administration of fenofibrate to 45 hyperlipidemic patients. The effects of 23 factors of physical examinations, clinical laboratory test values, and the background of the patients on the elevation of liver function test values were analyzed by the logistic regression method. The increase of the values of liver function tests was found to be correlated with BMI, serum levels of triglycerides, and ALP before therapy, especially sex, serum gamma-GTP level before therapy and reduced serum triglyceride levels. These results suggest that caution must be exercised to avoid liver dysfunction in patients treated with fenofibrate. PMID- 11857960 TI - [Development and application of an Internet-based educational system to share pharmaceutical case reports between pharmacists]. AB - For pharmacists to enhance skill of pharmaceutical cares, it is necessary to learn many instructive clinical cases, which include drug-related incidences and checks of medication errors by pharmacists. However, these cases tend to be kept in each facility and there is no access to the cases for outside pharmacists. Therefore, we aimed to develop an internet-based educational system to gather such cases, append educational commentaries by university staffs, and share them between pharmacists. An exclusive site was set up on the world wide web (WWW) homepage of the Section of Biopharmaceutics, Graduate School of Pharmaceutical Sciences, Kyushu University (http://seizai.phar.kyushu-u.ac.jp) on November 2000. Browsing was limited to registered pharmacists. Any pharmacist can be freely registered. Cases were collected via WWW-forms in the site. After educational commentaries and elucidation were appended, the cases were distributed to the registered pharmacists by e-mails and WWW pages. Many applications for membership have been sent from around Japan to have more than 50 members per month, and many cases have been contributed. These cases were distributed weekly by e-mail magazines. The developed system enabled us to share many educational cases of drug-related problems all over the country. This system could contribute to pharmacists to enhance their skills of pharmaceutical cares. PMID- 11857959 TI - [Cycloaddition of 1-methyl-2(1H)-quinolones having an electron-withdrawing group at the 3 or 4-position with 1,3-butadiene derivatives]. AB - Cycloaddition of 1-methyl-2(1H)-quinolones with electron-withdrawing groups such as methoxycarbonyl, cyano, and acetyl groups, at the 3 or 4-position with 2,3 dimethoxy- and 2-(trimethylsilyloxy)-1,3-butadienes afforded stereoselectively phenanthridone derivatives under atmospheric and high pressures. Furthermore, regioselectivities of the cycloaddition of 3- or 4-substituted 2 (1H)-quinolones with 2-(trimethylsilyloxy)-1,3-butadiene were examined using MO calculation. PMID- 11857961 TI - [Surveillance of reasonable period of antibiotic administration to compromised hosts]. AB - For the purpose of prevention of hospital-acquired infection caused by antibiotic resistant bacteria, we examined a method to establish an appropriate time period for the administration of antibiotics to compromised hosts. Using these antibiotics we monitored patients who received instruction about the drug regimen in the Blood and Respiratory Diseases Department ward. We monitored a) third generation cephalospolins, b) Imipenem/Cilastatin, and c) antibiotics used against methicillin-resistant Staphylococcus aureus. When the antibiotics were administered over 14 days, pharmacists notified physicians of the current duration of administration using a confirmation form, and confirmed their future administration schedule. We examined the antibiotic usage regimen of all the patients in this ward before and after the confirmation form was adopted. Patients given the same antibiotics within 14 days significantly increased in percentage from 82% to 91% after the confirmation form was adopted (p < 0.05). The median duration of antibiotic administration decreased from 7 days to 5 days. The case with antibiotic administration for the longest duration was a patient with leukemia who received vancomycin for 116 days after adoption of the confirmation form. This patient died 4 days after his antibiotic was changed. Only 16% of the patients administered antibiotics in this ward were monitored for the duration of antibiotic administration after adoption of the confirmation form. When the pharmacists positively provided physicians with information on some patients concerning the prolongation of antibiotics administration, the number of patients administered antibiotics for less than 14 days significantly increased throughout this ward without interfering with the treatment of patients who required long-term administration of antibiotics. PMID- 11857962 TI - [On psychology of the elderly--focusing on intelligence and senile dementia]. PMID- 11857963 TI - [Long-term care insurance system in Japan]. PMID- 11857964 TI - [Experience of long-term care insurance law for one year]. PMID- 11857965 TI - [Changes of care status before and after the introduction of the long-term care insurance in Japan--results of care status before the long-term care insurance]. PMID- 11857966 TI - [Change of comprehensive geriatric function and QOL of community-dwelling elderly in Japan by the implementation of long-term care insurance]. PMID- 11857967 TI - [Community care for housebound elderly after the long-term care insurance system]. PMID- 11857968 TI - [Proposal for measuring quality of care and perspective for practical use]. PMID- 11857969 TI - [Free radical theory of aging]. PMID- 11857970 TI - [Apoptosis and aging]. PMID- 11857971 TI - [Vascular regeneration]. PMID- 11857972 TI - [Neural stem cells: isolation and therapeutic application]. PMID- 11857973 TI - [Regeneration of cartilage/bone defects by growth/differentiation factors]. PMID- 11857974 TI - [Abdominal surgery in patients aged over 90 years old at a provincial general hospital]. AB - It has been commonly accepted that age itself is never an absolute contraindication for surgical treatment. Some many problems could be solved through surgical intervention, we studied perioperative status, postoperative morbidity and mortality, and short-and-long-term outcomes after abdominal surgery in a series of patients 90 year old or older, operated at a provincial General Hospital. Seven patients were treated using elective surgical procedures, while 6 had emergency surgery. The incidence of postoperative morbidity was fairly high after both elective and emergency operations. Major complications occurred in one patient who died on the 20th postoperative day, due to multiple organ failure. One case of hospital death occurred when a patient died on the 240th day after a gastrectomy. There was long-term survival of over five years occurred among the patients in the study who had reviewed elective operations for malignant lesions. No definite relation was revealed between the risk score and post-operative morbidity or mortality. No evident change in performance status was found even aged patient who underwent abdominal surgical procedures. These results indicate the need for more meticulous consideration, including more precise decisions regarding the indication for surgical intervention, and more intensive perioperative management, in order to secure more favorable therapeutic outcomes and quality of life for high-risk patients over 90 year of age. PMID- 11857975 TI - [Statement of initial therapy of elderly patients with hypertension in Wakayama prefecture]. AB - In order to evaluate the characteristics of initial therapy for elderly hypertensive patients in Wakayama prefecture, a case-card survey was performed in February 1997. The investigation consisted of blood pressure when starting therapy, initial physical examination and laboratory tests, and the initial drug therapy, and it's effects 3 months later. The initial physical examination and laboratory tests were the diagnostic procedures for determining the presence of target organ damage, they included electrocardiogram (ECG), chest X-rays, echocardiogram (UCG), funduscopy and urinalysis. Data of 7,647 cases from 156 facilities were obtained. These cases were divided into 2 groups, a non-elderly group (under 65 years, 3,396 cases) and an elderly group (65 years or over, 4,012 cases). The blood pressure at the start of pharmacological treatment was 174.9 +/ 17.4/93.7 +/- 11.0 mmHg (mean +/- SD) in the elderly group, and 170.6 +/- 17.8/98.6 +/- 11.2 mmHg in the non-elderly group. Systolic blood pressure in the elderly group was higher than in that the non-elderly group and diastolic blood pressure was lower in the elderly group than that in the non-elderly group. Cases of ECG (98.3% vs 71.3%; the non-elderly group vs the elderly group), chest X-ray (86.5% vs 65.2%), UCG (27.1% vs 23.7%), urinary test (96.0% vs 69.3%), examination of ophthalmic fundi (27.0% vs 24.3%), were much lower in the elderly group than in the non-elderly group. The rate of positive findings of left ventricular hypertrophy by ECG criteria (24.8% vs 20.3%), cardiomegaly by chest X p (35.3% vs 26.6%), proteinuria by urinary test (14.8% vs 12.9%) was lower in the elderly than non-elderly. Details of drug use in non-elderly vs elderly were as follows; calcium antagonists (47.3% vs 51.9%), angiotensin converting enzyme inhibitor (ACE-I) (14.2% vs 12.2%), diuretics (6.3% vs 10.2%), beta-blockers (11.2% vs 4.7%), alpha-blockers (1.4% vs 1.3%), others (1.5% vs 2.4%), multiple (13.7% vs 12.5%), undefined (4.2% vs 4.7%). Sufficient hypotensive effects were obtained in 61.5% of the non-elderly, and 68.5% of the elderly. Initial physical examination to assess target organ damage was lower in the elderly than the non elderly. It was thought necessary to be corrected. The incidence of main antihypertensive drugs in the elderly were Ca antagonists, ACE-I and diuretics. Treatment trends met the Japanese guideline on treatment of hypertension in the elderly. PMID- 11857977 TI - [Changes in nutritional status and nutritional intake among elderly residents of a community over a 10-year period: the Hisayama Study]. AB - The purpose of this study was to investigate the changes in nutritional status and nutrient intake among 387 elderly residents (> or = 60 y.o.) of Hisayama Town. A dietary survey was carried out at baseline in 1988 and repeated at the end of the follow-up period in 1998. During the 10-year follow-up period, mean values of body height and weight, and body mass index significantly decreased in both sexes. Likewise, blood hemoglobin and serum total cholesterol levels significantly decreased, while serum albumin and HDL-cholesterol levels significantly increased. Among dietary factors, mean energy intake significantly decreased due mainly to a decrease in the amount of carbohydrate intake, but no significant changes in the amount of protein and fat intakes were observed. The percentages of the Recommended Dietary Allowance for all nutrients eaten except for iron changed at the levels above 100%. The percentage of energy derived from carbohydrate significantly decreased, while those from protein and fat significantly increased. In conclusion, among community-dwelling healthy elderly people, mean energy intake significantly decreased due to a decrease in the amount of carbohydrate intake, and the fat intake relatively increased, with advancing age, but the blood lipid profile remained unchanged. PMID- 11857976 TI - [Effect of alfacalcidol on bone and calcium metabolism in elderly women]. AB - Oral administration of active vitamin D3 can reduce the loss of bone mass and the incidence of fractures in patients with osteoporosis in Japan. We conducted a prospective study to confirm the effects of 1 alpha(OH)D3 (Alfacalcidol, Alfarol Chugai Tokyo) on bone and calcium metabolism in elderly women with osteoporosis. Enrolled in the present study were 16 elderly osteoporosis women aged 72.6 +/- 4.5 years to whom 1 microgram of 1 alpha(OH)D3 was administered daily. Fasting blood and urine were obtained at baseline, 1 week, 4 weeks, 12 weeks and 24 weeks after the treatment. Monitored parameters were vitamin D metabolites, intact-PTH, bone alkaline phosphatase (BAP), osteocalcin (OC), deoxypyridinoline (DPD) and pyridiuium crosslinked type I collagen telopeptides (CTx). Serum 1 alpha, 25(OH)2D and PTH levels were significantly increased (p < 0.01) and decreased (p < 0.05), respectively at 1 week after commencing administration. There was a significant decrease of DPD (p < 0.05) at 12 weeks after commencing administration compared to the baseline levels. Serum levels of BAP and OC were found elevated at 1 week, and decreased at 12 weeks. In conclusion, the present study clinically confirmed that 1 alpha(OH)D3 stimulates bone formation in vitro. Long-term administration of 1 alpha(OH)D3 indirectly suppressed bone resorption through the suppression of parathyroid function in the elderly. PMID- 11857978 TI - [Relation between destination on discharge in the elderly patients and comprehensive geriatric assessment at admission in the ward of the Geriatric Department of University Hospital]. AB - To determine the factors related to the destination on discharge from the geriatric ward of Nagoya University Hospital, we analyzed the relationship between the scores of comprehensive geriatric assessment at admission and the destination in patients who had dwelled in home. The scores of basic activity of daily living (Barthel index), instrumental activity of daily living (Lowton scale), and cognitive function (Mini-Mental State Examination) were significantly lower in the patients who moved to institutions than those in the patients who returned home. The proportion of disabilities in all items, except eating, in the Barthel index, and all items but washing in the Lowton scale were significantly higher in patients who moved to institutions than in patients who returned home. Space orientation, calculation, and drawing in the Mini-Mental State Examination were related to the destination. In items for social life communication and group behavior were related to the destination. In multiple logistic regression models, it was suggested that activity of daily living, specifically independence of excretion, and ability in communication were significantly related to the destination on discharge. PMID- 11857979 TI - [Prospectives in proteomics]. PMID- 11857980 TI - [Structural quantification in osteoarthritis of the knee using power spectral analysis]. AB - An exact diagnosis of osteoarthritis is very important to prevent deterioration of ADL (activities of daily living) of the elderly. However, it is very difficult to diagnose the stage of osteoarthritis, and only a few indices for structural quantification of osteoarthritis have been reported. The purpose of this research is to investigate a new index for the bone structure in osteoarthritis. We examined right knees of 41 women aged from 34 to 85 years. Directivity index (DI) is a new index of bone structure calculated by directivity of the power spectrum from radiographs of tibial bone using fast Fourier transform (FFT). DI was obtained by subtracting the integral power value at 0 and 90 degree directions on the x-y plane of the two-dimensional power spectrum of tibial bone from the integral power value at a direction of 45 degrees. A significant relationship between the state of the knee joint by X-ray inspection and DI was indicated by evaluation of the correlation coefficient. However, no significant relationship was found between the state of the knee joint by X-ray inspection and the first moment of the Fourier power spectrum or the fractal dimension. There is a possibility that DI can estimate slight deformation of bone structure in osteoarthritis. In the future, we will apply DI to the prevention of osteoarthritis and a deterioration of ADL in the elderly. PMID- 11857981 TI - [A case of intracoronary stent implanted for acute myocardial infarction in an elderly patient with idiopathic thrombocytopenic purpura]. AB - A 68-year-old woman with idiopathic thrombocytopenic purpura (ITP) was admitted to our hospital with acute myocardial infarction on 7 February 1999. She had been treated since 1991 for mitral stenosis and regurgitation, atrial fibrillation due to mitralism, diabetes mellitus, hypertension, hyperlipidemia. Chest radiograph on admission showed cardiomegaly with congestion and cardiothoracic ratio was 63%. The platelet count on admission was 22,000/microliter, but she did not have petechia or purpura. Urgent coronary angiography revealed total occlusion in segment 7, and 13 and 75% stenosis in segment 4PD, 9 and 10. Subsequently, direct percutaneous transluminal coronary angioplasty (PTCA) was performed in segment 7. Dissection occurred during the intervention, and a coronary stent was implanted, we started heparin infusion and medication with ticlopidine hydrochloride as post stenting therapy after the intervention, and there was no bleeding tendency. PMID- 11857982 TI - [An elderly hypertensive patient with ocular bruits and angiographically confirmed stenosis of intracranical internal carotid arteries]. AB - The patient was 47 years old on his first visit in 1969 and has been treated at our outpatient clinic until now, at the age of 79. The initial blood pressure was 164/98 mmHg without abnormalities on electrocardiogram or urinalysis. He was followed-up for 8 years with non-pharmacologic treatment, which was eventually changed to antihypertensive drug treatment. At the age of 71 he was admitted because blood pressure increased to 210/110 mmHg, with marked fluctuation. At admission bilateral ocular (right < left) and carotid bruits (right < left) were detected. At the age of 75 he had cerebellar infarction with reversible neuro logic deficits. At the age of 76 he was again addmitted because the labile hypertension was difficult to control. Cerebral angiography which was performed to clarify the relationship between labile hypertension and cerebral ischemia revealed significant bilateral stenosis at the portion of the cavernous sinus in the internal carotid arteries. The stenosis was more remarkable on the left side than the right side. Based on the angiographic findings we considered the ocular bruits to be not a murmur of augmentation flow but a stenotic murmur. PMID- 11857983 TI - [An elderly case of cytomegalovirus enterocolitis associated with a malignant tumor]. AB - An 83-year-old man was given a diagnosis of left parotid cancer in our hospital in November 1997. He refused to undergo a surgical procedure because of his advanced age. Therefore chemotherapy and radiotherapy were used. Chemotherapy with CAP (Cyclophosphamide, Adriacin, and CDDP) was conducted on 6 occasions between December 1997 to July 2000. Prednisolone (15 mg) was administered daily from July 15, 2000. The patient started suffering from diarrhea on August 2, 2000. As the patient also began to suffer high grade fevers and stomachaches, he was admitted on a diagnosis of acute enterocolitis. He had bloody stool on August 11. On emergency colonoscopy, an ulceration with bleeding was located in the lower rectum. The biopsy specimen revealed intranuclear inclusion bodies and positively staining cells for monoclonal antibody to cytomegalovirus through the immunohistochemical technique, and it was diagnosed as cytomegalovirus enterocolitis. He was treated with ganciclovir. One month later, his clinical symptoms had improved. Cytomegalovirus enterocolitis is an opportunistic infection, so immunocompromised hosts (such as cancer patients, patients using immunosuppressants, old people) have a greater probability of contracting cytomegalovirus infection. A ganciclovir is an effective treatment. A cytomegalovirus enterocolitis should considered in the differential diagnosis of enterocolitis, when alimentary symptoms like diarrhea or bloody stool are found in immunocompromised hosts. PMID- 11857984 TI - [Clinical relevance of the Streptococcus milleri group in head and neck infections]. AB - Streptococcus constellatus, S. intermedius, and S. anginosus, the 3 species of the S. milleri group, form part of the normal flora commonly found in the mouth, throat, and gastrointestinal and genital tracts. This group has become known as an important pathogen in infections and abscesses, but data on the anatomical distribution of these species is lacking in relation to clinical significance. We obtained 275 strains of the S. milleri group from different departments at our hospital over the last 3 years, including 54 strains from dental surgery, 47 from internal medicine, 44 from otolaryngology (head and neck), 43 from surgery, 32 from gynecology, 17 from urology, 16 from dermatology, 11 from brain surgery, 6 from pediatrics, 3 from orthopedics, and 2 from opthalmology. The 44 strains from head and neck were found in 42 patients,--23 with primary infection and 19 with secondary infection induced by cancer treatments. The primary infection group included 4 deep neck abscesses, 1 peritonsillar abscess, 5 tonsillitis, 4 paranasal sinusitis, 3 congenital aural fistula infections, 2 dental infections, 2 paranasal sinus cysts, 1 supprative parotitis, and 1 postoperative wound infection. The secondary infection group included 7 postoperative wound infections, 3 postoperative pulmonary infections, 3 laryngitis and pharyngitis, 3 terminal pneumonias, and 3 infections of the local recurrence site. The S. milleri group was the only isolated organism in 13 cases (56.5%) of primary infection and in 5 (26.3%) of secondary infection. Among other organisms from the primary infection group, no so-called major pathogens were found. Antimicrobial susceptibility tests of the S. milleri group showed that 50% were resistant to CCL and 33% to CTM. ABPC, CPDX, and CFDN were also found to be less sensitive, although no resistant strains were detected. To adequately culture the S. milleri group, incubation in air containing carbon dioxide or in an anaerobic atmosphere is required, and differentiation of the 3 requires biochemical reactivity tests. Since not all facilities use identical techniques in routine bacteriological examination, a considerable number of the S. milleri group could be missed in unknown species of alpha-,beta-, and gamma-streptococci and culture-negative cases. With antibiotics now being used widely, normal flora such as the S. milleri group may have become an important pathogen in head and neck infections due to an imbalance between organisms and host defense. PMID- 11857985 TI - [Clinical investigation of acute sensorineural hearing impairment due to rifles]. AB - This study examined 53 patients (74 ears) with acute sensorineural hearing impairment due to rifles. After patients were admitted, 16 were treated by steroid alone and 37 by steroid and PGE1 through intravenous drip injection. The therapeutic ratio was 19% and the recovery ratio 66% with no significant difference seen between treatment types in hearing improvement. Forty-two patients were treated within 7 days of injury and 32 8 or more days after injury. The therapeutic ratio was 21% and the recovery ratio 78% among patients treated sooner and a significant difference was seen between in hearing improvement early and later (8 or more days) treatment. We studied average hearing improvement between pre- and post treatment across the frequency spectrum. The average hearing improvement of 500 Hz and 1 kHz was significantly high compared to that of 8 kHz. This suggests that impairment of middle-pitched tones is improved more easily than high-pitch tones in hearing injuries due to rifles, and that early treatment after injury is extremely important. PMID- 11857986 TI - [A case of 9p-deletion syndrome with congenital median nasal fistula]. AB - We report a case of 9p-syndrome with congenital median nasal fistula in a boy born to a 28-year-old mother as the second child by normal delivery. The fistula opened at the base of the bridge of the nose and ran between the nasal septum cartilage to the anterior cranial fossa. A frontal craniotomy and transcolumellar skin incision were conducted to extirpate the fistula. In the 10 months since, no fistula has recurred. PMID- 11857987 TI - [Swallowing function after near-total laryngectomy, cricohyoidoepiglottopexy (CHEP), and cricohyoidopexy (CHP)]. AB - We studied postoperative swallowing in 4 patients undergoing CHEP and 1 undergoing CHP. Swallowing was obtained by intense swallowing rehabilitation since only 1/4 of the larynx remained after near-total laryngectomy. Our swallowing rehabilitation program is detailed in this paper. The improvement of swallowing is classified into 3 stages. In stage I, volus directly intrudes into the trachea. In stage II, volus stagnates between laryngeal inlet and tracheal stoma. In stage III, volus directly flows through the esophageal inlet. Stage III indicates that rehabilitation is almost completed. Stage I is shortest at 2 to 14 days and Stage II longest at 7 to 80 days. The MTF (Method, Time, Food) score described by Fujimoto et al was used to analyze swallowing. Three cases following CHEP showed high scores shortly after the introduction of rehabilitation and reached the maximum score at discharge (15 points = normal swallowing). At present, these 3 patients are satisfied with swallowing and enjoy a good quality of life. In 2 other cases (1 CHEP and 1 CHP), both had a wide laryngeal inlet and still have some difficulty with liquids. Further modification of the surgical technique is needed especially for CHP. PMID- 11857988 TI - Hype? What hype? PMID- 11857989 TI - IMRT update. PMID- 11857990 TI - Preparing a business justification for going electronic. AB - Exponential advances in the technology sector and computer industry have benefited the science and practice of radiology. Modalities such as digital radiography, computed radiography, computed tomography, magnetic resonance imaging, ultrasound, digital angiography, and gamma cameras are all capable of producing DICOM compliant images. Text can likewise be acquired using voice recognition technology (VRT) and efficiently rendered into a digital format. All of these digital data sets can subsequently be transferred over a network between machines for display and further manipulation on workstations. Large capacity archiving units are required to store these voluminous data sets. The enterprise components of radiology departments and imaging centers--radiology information systems (RIS) and picture archiving and communications systems (PACS)--have thus undergone a transition from hardcopy to softcopy. When preparing to make transition to a digital environment, the first step is introspective. A detailed SWOT (strengths, weaknesses, opportunities and threats) analysis, with a focus on the status of "electronic preparedness," ensues. The next step in the strategic planning process is to formulate responses to the following questions: Will this technology acquisition provide sufficient value to my organization to justify the expense? Is there a true need for the new technology? What issues or problems does this technology address? What customer needs will this technology satisfy today and tomorrow? How will the organization's shareholders benefit from this technology? The answers to these questions and the questions that they in turn generate will stimulate the strategic planning process to define demands, investigate technology and investment options, identify resources and set goals. The mission of your radiology center will determine what you will demand from the electronic environment. All radiology practices must address the demand of clinical service. Additional demands based on your mission may include education and research. The investigation of options is probably the most time consuming portion of the analysis. It is in this stage where the system architecture is drafted. Important contributions must be solicited from your information technology division, radiologists and other physicians, hospital administration and any other service where the use of imaging technology information is required and beneficial. Vendors and consultants can be extremely valuable in generating workflow diagrams, which include imaging acquisition components and imaging display components. A request for proposal (RFP) may facilitate this step. A detailed inventory of imaging equipment, imaging equipment locations and use, imaging equipment DICOM compatibility, imaging equipment upgrade requirements, reading locations and user locations must be obtained and confirmed. It is a good idea to take a careful inventory of your resources during the process of investigating system architecture and financial options. An often-ignored issue is the human resource allocation that is required to implement, maintain and upgrade the system. These costs must be estimated and included in the financial analysis. Further, to predict the finances of your operation in the future, a solid understanding of your center's historical financial data is required. This will enable you to make legitimate and reasonable financial calculations using incremental volumes. The radiology center must formulate and articulate discrete clinical and business goals for the transition to a digital environment that are consistent with the institutional or enterprise mission. Once goals are set, it is possible to generate a strategic plan. It is necessary to establish individual accountability for all aspects of the planning and implementation process. A realistic timetable should be implemented. Keep in mind that this is a dynamic process; technology is rapidly changing, as are clinical service demands and regulatory initiatives. It is therefore prudent to monitor the process, make appropriate revisions when necessary and address contingencies as they arise. PMID- 11857991 TI - Learn from experience: insights of 200+ PACS customers. AB - This article is based on a PACS study published in June 2001 by KLAS Enterprises. The study offers unique insight into the performance, underlying technology, product depth and breadth, and "real world" issues facing 10 of the leading PACS vendors and their clients as rated at more than 200 of their client sites by CIOs, department directors and vendor executives. The driving force behind this report reflects the heightened interest in PACS systems in general and PACS' contributions to the benefits associated with an electronic medical record. This research was sponsored by 10 healthcare provider organizations (not vendors), ranging from a 150-bed acute-care hospital to a seven-hospital IDN. Most of the data in the report focuses on the use of PACS in the radiology department, since comparatively few sites are using PACS systems in other departments, such as cardiology or pathology. The participating vendors supplied their customer lists, which contributed to the majority of clients surveyed. Clients surveyed in the study vary widely in size, ranging from 50 to 2,000 beds, with about 46 percent of those surveyed having a hospital size of more than 400 beds. The customers represent a broad cross-section of PACS clients, ranging from 10 percent to 95 percent filmless, with large-scale users making up the majority of respondents. Each of the customers were asked to numerically rate their vendor on 28 aspects of performance, as well as to answer 12 questions relating to customer satisfaction. Based on study results, success with PACS can be distilled into some basic principles. There are other issues that will help with a successful PACS selection and implementation, but the following list should help anyone get started. If you are able to focus on and accomplish these few things, you stand a better chance of building and meeting your business case. Pre-sell your PACS concept to the radiologists, technicians, IS/IT staff supporting it and to the referring physicians. Make your system user-friendly. Select a vendor that has a history of partnering with their clients. Find a vendor and system that you can afford and that provides enterprise-wide distribution and access to images. Move quickly from printed film to digital images. Know yourself. Know your goals, Do your homework. Ask for help. PMID- 11857992 TI - Screening imaging procedures. PMID- 11857993 TI - Screening imaging procedures: a good idea or not? PMID- 11857995 TI - Web distribution of diagnostic images requires new security efforts. PMID- 11857994 TI - The gift of employee dissatisfaction. AB - Through an employee survey administered at Kaweah Delta Health Care District (KDHCD) in Visalia, Calif., several sources of dissatisfaction were noted, including communication, equipment, staffing and rapid growth. Perceiving no real movement toward resolving these issues, employees vented their frustrations to administration. As director of imaging services, I enlisted the help of two inside consultants, KDHCD's director of education and the director of the Employee Assistance Program (EAP). We initiated a process that is ongoing, to move the department toward working together as a team to solve problems within their control. We three directors decided to work with a leadership group to: assess the history of the department clarify the current reality create a vision of the future learn the Covey Habit 4, "Think Win-Win" capture agreements that lead staff and managers to work in self-motivated, self-directed work teams have the director of education present the work to the staff at large, and encourage the leadership team to continue to learn tools that would help the group to improve. The two inside consultants held a series of training meetings with the leadership group of 15, which included a staff member from each modality, site and support service. Participation was optional, and all who were asked agreed to participate. The meetings were held weekly for four weeks for two hours before regular work hours. At the conclusion of the training, the group agreed to continue to meet weekly. After the first four meetings, a summary of the training was presented at a meeting of the full imaging staff plus the vice president of professional services at KDHCD. Through this program, imaging services staff members at KDHCD have achieved an increased sense of cohesion in the group, learned that we have control over some things and not others, and are learning to hold each other accountable with kindness. We are giving each other the benefit of the doubt. We have not achieved perfection, however we have renewed hope for a brighter future. PMID- 11857996 TI - Strength from adversity. PMID- 11857997 TI - A radiography program can be part of the solution. PMID- 11857998 TI - Ask Sirenhead. Portable problems. PMID- 11857999 TI - Case of the month. Distracted by the facts. PMID- 11858000 TI - The new ABCs of AV block. A revised classification to remove the mental block from recognizing AV block. AB - This revised classification system is designed to etch the indicators of a block into your memory. You only have to remember that the degree is the number of atrial beats getting through to the ventricles (all, some or none) and the type is where the block occurs (AV node or septum). Once you learn this system, simply practice interpreting block rhythms (see "AV Block Case Studies," p. 33). This revised block classification brings bundle branch blocks into the fold, so to speak, allowing you to focus on the block's anatomical location (see Figure 14, left). It also reinforces the fact that whenever we use the word block, we're talking about AV block. So how do you put it all together to use this new classification system? Use your usual rhythm strip analysis of rate, rhythm, P, PR, QRS, ST, T, U and summary to do the following: 1. Note any PR or QRS abnormalities. 2. Count the number of atrial and ventricular beats in a six second ECG strip. If the ratio is 1-to-1 and the PR and QRS are normal, then there's no block present. If the ratio is 1-to-1, the PR is prolonged and the QRS is narrow, it's a first-degree, type 1 block. If the ratio is 1-to-1, the PR is normal and the QRS is wide, it's a first-degree, type 2 block. If the ratio is greater than 1-to-1 (such as 2-to-1, 3-to-1, etc.), but some of the beats are conducted, it's a some (second-degree) block. If the PR prolongs and the QRS is narrow on the conducted beats, it's considered a second-degree, type 1 block (Mobitz 1 or Wenckebach). If the PR is normal and the QRS is wide on the conducted beats, it's a second-degree, type 2 block (Mobitz 2). If none of the beats appear conducted (the PR interval is completely variable), it's a none (third-degree) block. If the QRS complex is at a normal width, then it's probably a third-degree, type 1 block. And if the QRS complex is wide, it's a third degree, type 2 block. As with anything in medicine, no system is perfect. However, through using this revised classification of AV block--based on the heart's anatomy--you can permanently retain the features of the various AV blocks in your memory. Now that you're feeling more comfortable with block rhythms, examine Figure 15. This is a patient in sinus tachycardia who has received adenosine. Notice how the patient goes from sinus tachycardia to first-degree, type 1 to second-degree, type 1 to second-degree, type 2 and back again. It's a very rare strip that you can use to test your ability--and the ability of your crews--to interpret block rhythms. PMID- 11858001 TI - Strategic deployment. Two decades of experience provide important lessons on how to deploy emergency resources. AB - When it comes to deployment, one size (approach) doesn't fit all. Progressive public safety and EMS systems will recognize that a successful plan can thoughtfully balance system performance and human needs. Inventive people who learned how to do things better, cheaper and faster built America. Providing optimum system coverage, improved response times and quality care for patients fits the same mold. Sophisticated and accountable EMS systems readily accept this as part of their mandate. PMID- 11858002 TI - 200 city survey. JEMS 2001 annual report on EMS operational & clinical trends in large, urban areas. AB - This year's survey offered examples of evolving partnerships between the public and EMS providers with a growing number of systems implementing PAD programs. The apparent influence of a communication center's managing agency on prioritization strategies is concerning. However, further study is needed. EMS managers must pay careful attention to comm center practices and technology to ensure their ability to support response prioritization and the efficient management of EMS resources. The small reduction in the use of hot response (lights and siren) to every request for service is disappointing in light of medical literature and position statements that condemn this practice. Resource response can be safely prioritized using today's EMD protocol systems. Prioritization and changing response [figure: see text] time requirements to address impending revenue and service demand changes will require additional standardization of methodologies and reporting of response times to relate this measure to other system performance indicators (e.g., patient morbidity/mortality, cost, customer satisfaction, etc.). The future presents a difficult road for system administrators. However, the adoption of a growing number of information management tools and changes in procedures and dispatch processes offer potential solutions. The increased use of hand-held computers or personal digital assistant (PDAs) to gather and provide information and the almost universal use of CAD will aid providers in performing the research necessary to change response time performance requirements, improving EMS system efficiency. Use of this technology will also likely improve patient care and reimbursement through more timely and accurate reporting and analysis. The medical director's role will be critical to ensuring potential changes don't compromise patient care. Obtaining a better understanding of how much time can safely elapse between the time of the 9-1-1 call and when patient-care activities commence will be an important component of future strategies. More sophisticated EMD, CAD and AVL technologies will also play an indispensable role in reforming system design and daily operations. In light of the events of Sept. 11 and events yet to occur, EMS managers and providers face significant operational challenges. Overcoming these challenges will require leadership, a willingness to question and change tradition and the ability to cope with the discomfort of changing demands and uncertainty. Maintaining the hard-fought successes of EMS will increasingly require more imagination and the willingness of current and future practitioners to study, develop and implement innovative approaches to addressing future requirements. PMID- 11858003 TI - Diagnosis: cancer. Understanding common cancer treatments, their side effects & appropriate EMS care. PMID- 11858004 TI - Terror or consensus? PMID- 11858005 TI - Rebirth of the university hospital: the Finnish strategy. AB - Like the academic hospitals worldwide, the Finnish university hospitals are under increasing economical pressures seeking ways to re-assess their traditional missions. It is strongly felt that new solutions within the field of evaluative sciences could bring added value to university hospitals. In this vein, university hospitals must take a leading position in assessing efficacy, (cost )effectiveness and, ultimately, cost-utility of both traditional and emerging clinical technologies. This will affect the research culture of university hospitals both profoundly and permanently. PMID- 11858006 TI - [The role of hospitals in the new century]. AB - The century could be described as an era of transformation for hospitals, from the refuge where the poor went to die to a high tech institution. Indeed, hospitals benefited not only from advances in diagnostic, therapeutic and preventive medicine, but also from the widest range of sciences and technologies. Most diseases can be eradicated and hospitals have largely contributed to their disappearance. Today, the picture could therefore be idyllic. But this would not take into account the wide inequalities between countries, and even within a country. The failure of the WHO message for 2000, "Health for all," is also the failure of the hospitals. I would simplify this presentation by making a distinction between three categories: OECD countries, less advanced countries and emerging countries. PMID- 11858007 TI - Implementing chronic disease management in the public healthcare sector in Singapore: the role of hospitals. AB - The public health care delivery system in Singapore faces the challenges of a rapidly ageing population, an increasing chronic disease burden, increasing healthcare cost, rising expectations and demand for better health services, and shortage of resources. It is also fragmented, resulting in duplication and lack of coordination between institutions. A disease management approach has been adopted by the National Healthcare Group (NHG) as a critical strategy to provide holistic, cost-effective, seamless and well-coordinated care across the continuum. The framework in the development of the disease management plan included identifying the diseases and defining the target population, organizing a multi-disciplinary team lead by a clinician champion, defining the core components, treatment protocols and evaluation methods, defining the goals, and measuring and managing the outcomes. As disease management and case management for chronic diseases are new approaches adopted in the healthcare delivery system, there is a lack of understanding by healthcare professionals. The leadership and participation of hospital physicians was sought in the planning, design and outcomes monitoring to ensure their 'buy-in' and the successful implementation and effectiveness of the program. The episodic diagnosis related group (DRG)-based framework of funding and subvention for healthcare, and the shortage of step-care care facilities, have been recognized by the Ministry of Health as an impediments to the implementation, and these are currently being addressed. PMID- 11858008 TI - Global development challenges and health care reform. AB - Changes in the role of the state and private sector are seen as central to success of many health care reforms. The article argues for a more focused "stewardship" function of governments in securing equity, efficiency, and quality objectives through more effective policy making (steering), regulating, contracting, and ensuring that adequate financing arrangements are available for the whole population. At the same time, the author argues a strong case for greater private participation in providing health services (rowing). The article reviews related reform trends in health care financing, generation of inputs and service providers. It concludes that reforms often fail, not because of a flawed technical design, but because of other factors. These include a lack of political commitment to change, resistance from vested stakeholders who fear loosing some of their existing benefits, and a failure by policymakers to translate successful aspects of the reforms into something visible that ordinary people and the public can see with their own eyes when next they use the reformed services. PMID- 11858009 TI - Healthcare architecture in metamorphosis--observations in Hong Kong's heuristic experience. AB - Healthcare Architecture in Hong Kong is in an on-going process of metamorphosis in response to the social, economical and technological developments in the territory. In the process of transformation, universal problems like obsolescence, growth and expansion, and advances in science and technology as well as problems unique to Hong Kong like population growth, scarcity in land supply and high density development all call for special solutions. With the turn of the century, new forces of change have also begun to take shape, and in anticipation of the hyper-turbulent changes ahead, we need to shift our paradigm to allow revolutionary new perspectives and innovate, shape and create the future healing space which is sustainable, adaptable, flexible and humane. PMID- 11858010 TI - Developing a holistic view and approach to health care. Rene Sand Lecture. International Hospital Federation Congress--May 2001. AB - One of the significant features of Hong Kong is its public healthcare system, which provides universal, easily accessible and heavily subsidized medical care for all. The governing policy has been to ensure that no one is denied adequate medical care because of insufficient means through the provision of a comprehensive range of healthcare services funded from the public purse at extremely low fees to users. Over the past decade, the public hospital sector has been able to provide efficient and quality health services comparable to those of the industrialised nations. However, the reality today is that the public hospitals are facing many challenges that must be overcome and ways must be sought to mitigate the growing strains on the healthcare system and rebalance the healthcare economics. The recently published Consultation Document on Healthcare Reform, by Dr E K Yeoh, Secretary for Health & Welfare, proposes the introduction of a comprehensive range of short-term and long-term initiatives to revitalize and transform the healthcare system into a community-focused, patient-centred and knowledge-based integrated healthcare service with improved interface between the public and private sectors. PMID- 11858011 TI - Reform of the health care service system in Israel 1995-2000. AB - In January, 1995, a national health insurance law was enacted which allowed for the registration of every resident with one of the four public health funds. These funds provide medical services directly to their insured members, or purchase them from public or private hospitals or other health service providers. A basket of medical services was defined which guaranteed readily available comprehensive health care throughout the country. As part of the reform, a number of personal participation charges were established: a nominal fee for medications, and a service charge for a limited number of medical services (visits to specialist doctors, periodic fee for mother and child clinics, participation in the cost of chronic nursing hospitalization). In spite of the Government's commitment to finance the basket of services, the budget is not sufficient to cover costs, and there is an inherent shortfall deliberately introduced into the system by the Ministry of Finance. Since the introduction of the Law the sources for financing the basket of services have been eroded by 16%. Complementary health insurance programs are being developed by the health funds as well as by private insurance companies. The national expenditure on health is some 8.4% of GDP, with an average annual expenditure of $1,384 per person. 72% of health expenditures are public and 28% private. Life expectancy is continually rising, and stood at 75.9 years for men and 80.1 years for women in 1999. Infant mortality is decreasing: in 1996 the rate was 6.3 per 1,000 live births. There is an ever increasing use of new technologies, even though at the same time there seems to be somewhat of a decrease in customer satisfaction. The achievements of the reform are considerable, but there is need for improvement in the areas of budgeting, return of the employer parallel tax, and in other areas if these achievements are not to be undone. PMID- 11858013 TI - [Transmeatal direct myringoplasty]. AB - A surgical method of transmeatal direct myringoplasty (TDM) was performed in 30 patients. Out of those, 21 patients were previously treated for chronic otitis media, 4 patients were with dry perforations of neomembrane following myringoplasty, and 5 patients were with blasttraumatic ruptures of the eardrum. Fascia of the temporal muscle was used in the defect reconstruction. Graft healing and closure of the eardrum defect were observed in all patients 3 months after the surgery. Postoperative improvement of hearing in an interval of 15-20 dB was found in 21 patients. Milder sensorineural hearing damage was found in 1 patient, while in the others the hearing was on the preoperative level. Simplicity of the method, minimal surgical procedure and favorable postoperative results were the reason why the authors recommended TDM as the method of choice in the reconstruction of the eardrum defects caused by an inflammation or blasttraumatic damages of the eardrum. PMID- 11858012 TI - [Complications in omentopexy and personal experience with 100 omentomyelopexies]. AB - Abdominal and general complications of omentomyelopexies are seldom seen, and were mostly presented as the case reports. The aim of this study was to present in a comprehensive analysis of the data acquired from literature the reported omentopexy complications, and to compare them with our own results of the application of the omental flap with left gastroepiploic blood vessels. Prospective clinical research included the first 100 operated patients with injuries on different spinal cord levels. In all patients the omental flap was prepared by our original surgical technique and led to the site of the spinal cord lesion. In 6 of our patients we noticed seven complications and those were: three iatrogenic pneumothoraxes, two lesions of the spleen (without splenectomy), one pneumonia and one mild infection of the laparotomy wound. Prevalence of the complications related to the abdominal act of omentomyelopexy of 7% was significantly lower than in those presented in the literature with other right gastroepiploic artery omentopexies (20-79%). In our patients complications occurred earlier, lasted shorter and had no permanent implications for the patients' health. By using the omental flap with left gastroepiploic blood vessels it was possible to perform effective, relatively simple and safe omentomyelopexy for the injuries on all levels of the spinal cord. Such a mode of omental flap preparation with left gastroepiploic blood vessels could be applied for other omentopexies as well. PMID- 11858014 TI - [Diagnosis of subarachnoid hemorrhage]. AB - OBJECTIVES AND METHODS: A total of 115 patients with subarachnoid hemorrhage were retrospectively analyzed with the aim to evaluate the timing and reliability of the applied diagnostic procedures. In the group of 63 patients the reliability of CT as the diagnostic procedure was investigated, and CT scan was correlated with the clinical status of the respective patient. In the group of 79 patients with operatively treated cerebral aneurysm the angiographic and intraoperative findings were mutually compared and discrepancies were analyzed. RESULTS: During the first 48 hours after the hemorrhage, only 35.2% of all the angiographies were performed, so the final diagnosis was late in 64.8% of patients. Hemorrhage was diagnosed by CT in 87.3% of the cases and pronounced correlation existed between CT and the clinical status of the patient. Discrepancies between angiographic and operative findings existed in 16.4% of patients. CONCLUSION: CT should be the initial procedure for the diagnosis of subarachnoid hemorrhage. In nonmoribund patients the early angiography should also be performed. The main reason for delaying in diagnosis is the overdue transfer of the patients to the referral hospitals. PMID- 11858015 TI - [Symptoms of locally advanced non-small-cell lung carcinoma]. AB - A group of 50 patients with locally disseminated non-small-cell lung cancer was analyzed. The diagnosis was confirmed upon the analysis of the samples obtained by bronchoscopy or percutaneous needle biopsy of the lungs. CT scan of the thorax, as well as transesophageal echosonography of the mediastinum and ultrasound of the abdomen were performed for the evaluation of the disease dissemination. The aim of the study was to establish the main characteristics of lung cancer in locally disseminated disease with particular reference to the analysis of the symptoms. The majority of patients were aged between 56 and 60 years (n = 13; 26%). The majority was with planocellular cancer of central localization (n = 38; 76%). In 20 (40%) patients surgery was performed, and in 30 (60%) patients the surgery was finished with exploration. The most frequent symptoms were cough in 29 (58%), hemoptysis in 27 (54%) and chest pain in 24 (48%) patients. Frequency of the symptoms was multiplied in the patients with the advanced disease. Frequency of hemoptysis was equal in the group of patients who were operated and those with exploratory thoracotomy. PMID- 11858016 TI - [Diagnostic validity of radionuclide phlebography in the detection of clinically occult deep venous thrombosis in patients with thromboembolism]. AB - Diagnostic reliability of radionuclide phlebography (RNP) compared to contrast phlebography in the detection of deep veins' thrombosis (DVT) in patients with confirmed thromboembolism of the lungs (TEL) was evaluated. RNP. These findings were compared to contrast phlebography (CP), performed in the group of 25 patients in whom TEL was confirmed clinically, radiologically, scintigraphically and biochemically, but without clinical signs and symptoms of DVT. In 15 patients where RNP revealed unilateral DVT, CP finding was confirmed in all: in 6 (40%) on the left, and in 9 (60%) on the right side. In 10 patients with bilateral signs of DVT observed by scintigraphy, DVT finding was confirmed in 7 (70%), while in 3 (30%) patients, scintigraphic signs of DVT were falsely positive. In the segments of deep venous system, specificity of RNP in the detection of DVT in the lower leg was 60%, sensitivity was 100%, accuracy was 64% with 62% falsely positive findings. In the upper legs specificity was 79%, sensitivity was 100%, and accuracy was 86% with 24% falsely positive findings, while in the pelvis specificity was 87%, sensitivity was 100%, accuracy was 83% with 14% falsely positive findings. PMID- 11858018 TI - [Causes of frustration in soldiers during the period of adaptation to the military environment]. AB - The aim of this study was to determine the frustrations that prevent soldier's successful adaptation to the military environment in the first three months of military duty. The sample consisted of 200 adapted (group A) and 400 maladapted (group NA) soldiers. The applied instrument was the Adjustment questionnaire. It was concluded that the most important cause of frustration was the problem of establishing the interpersonal relationship of soldier and his comrades in the unit. The presence of the actual factor of decompensation was also important, in the sense that it could disqualify for military duty, temporarily or permanently, even a well adapted soldier. Commanding officers and comrades could positively influence the socialization of a personality, by helping the less sociably adaptable soldiers in their adaption to the military environment. PMID- 11858017 TI - [Reliability of plasmid profile analysis in the identification of epidemic strains of bacteria causing an outbreak of intestinal infections]. AB - The aim of the study was to determine the plasmid profile of isolates originated from the outbreaks of gastrointestinal tract infection in the attempt to identify the outbreak strains. Plasmid profile analysis was performed on 163 isolates; 82 isolates originated from foodborne outbreakes and 81 isolates originated from waterborne outbreakes or outbreakes spread by contact. Outbreak strain was identified in 1 (10%) of 10 foodborne outbreakes and in 7 (70%) of 10 waterborne outbreakes or outbreakes spread by contact. According to the obtained results, the method of plasmid profile analysis was more reliable in the investigation of outbreaks of gastrointestinal tract infection spread by water or contact compared to the outbreakes spread by food. PMID- 11858019 TI - [Work capacity evaluation in workers with arterial hypertension]. AB - The aim of this study was to show the results of working ability estimation of the workers with arterial hypertension. The estimation included 313 workers with arterial hypertension estimated by Disability Commission during 1999. Arterial hypertension was a leading diagnosis in 30.9% cause of working ability estimation. The majority of workers belonged to the 41-60 years age group and were with arterial hypertension with complications on the fundus and on the heart. In the examined group the majority of workers was declared as first category invalid (35.8%), while 32.3% of hypertensive workers had to change the working place. The majority of the first category invalids were among the hypertensive workers from textile and chemical industry, as well as among the professional drivers. The changing of the working place was more frequently suggested to the workers in textile and metal industry and in printing firms. These results could be explained by the influence of professional noxious agents on the onset and development of arterial hypertension. PMID- 11858020 TI - [Formulation of lithium carbonate tablets with various binding substances]. AB - The test results of lithium carbonate tablets, made of various binding substances (maize starch, gelatin, sodium carboxymethylcellulose 7pp, kollidon 30) were presented. The content of lithium carbonate, tablets weight variation, hardness, friability, disintegration, as well as the lithium carbonate dissolution rate were determined. The best properties were observed in tablets made with maize starch paste with the addition of starch in the internal phase. PMID- 11858022 TI - [The bioartificial liver]. PMID- 11858021 TI - [Evaluation of cermet fillings in abutment teeth in removable partial prostheses]. AB - The aim of the study was to describe the clinical process of setting the purpose filling on abutment teeth, after finishing the removable partial dentures. The aim was also to investigate the use of cermet glass-ionomer cement for the purpose filling in the abutment teeth for removable partial dentures, as well as to investigate the surface of the purpose filling. For the clinical evaluation of purpose filling slightly modified criteria according to Ryg's were used in 20 patients with different type of edentulousness. Changes occurring on the surface of purpose filling have been experimentally established by the method of scanning electron microscopy on the half-grown third molars in seven patients. It could be concluded that cement glass-ionomer was not the appropriate material for the purpose fillings in abutment teeth for removable partial dentures. PMID- 11858023 TI - [The importance and role of surgical methods in the treatment of ischemic cerebral disease]. PMID- 11858024 TI - [Endoscopic methods of treatment of non-variceal bleeding lesions in the digestive tract]. PMID- 11858025 TI - [Tear gases as incapacitating agents]. PMID- 11858026 TI - [A severe form of falciparum malaria associated with staphylococcal endocarditis]. AB - A case is presented of a patient, aged 56 years, with severe form of imported malaria caused by Plasmodia falciparum. Hyperparasitemia of erythrocytes > 30% was registered, and during the course of the disease CNS dysfunction, severe anemia, acute renal failure, disseminated intravenous coagulation with manifest hemorrhagic syndrome, icterus, enterocolitis, pneumonia and staphylococcal endocarditis were developed Due to hyperparasitemia and numerous complications, antimalarial drugs such as quinidine (1,200 mg/day) and artemether (160 mg/day) were administered parenterally. Infected erythrocytes were exchanged with 2.5 litres of healthy erythrocytes suspension. Hemodialysis was also performed as well as nine-week antistaphylococcal therapy. During the treatment preparation of deplasmated blood, concentrated thrombocytes, fresh frozen plasma, cryoprecipitates, human albumins and immunoglobulins were applied, along with the correction of electrolytic dysbalance, administration of diuretic, cardiotonic, antiarrhythmic, anxiolytic, antipsychotic and antidepressive drugs. Two months after the admission the patient was released from the Clinic in good condition, with normal clinical-laboratory findings. PMID- 11858027 TI - [Laza K. Lazarevic (1851-1891)--part 1]. PMID- 11858028 TI - [Preoperative computed tomography of temporal bones and its use in selecting patients with sensorineural deafness for cochlear implantation]. AB - Cochlear implantation (CI) involves the insertion of electrode systems in the inner ear in order to restore hearing in patients with sensorineural deafness. Positive CI results are closely related with careful selection of candidates. Preoperative computed tomography (CT) and its results are decisive in this selection. Temporal bone CT was made in 40 patients aged 1 to 57 years who had sensorineural deafness. The following changes were detected: malformations of the inner ear, the inner auditory canal, meningititis-induced cochlear obliteration, post-traumatic temporal bone changes, local auditory ossicle malformation, inflammatory middle ear changes, the high position of the jugular vein bulb, and its diverticula. Temporal bone CT plays an important role in selecting candidates for CI and circumvents potential difficulties and complications at CI. PMID- 11858029 TI - [Clinical experience with positron emission tomography with 2-fluorine, 18F-2 deoxy-d-glucose used for the diagnosis of malignant breast neoplasms]. AB - The aim of the study was to evaluate sensitivity and specificity of positron emission tomography (PET) and 18F-fluorodesoxyglucose (18F-FDG) in breast cancer diagnosis and to assess tumor dissemination. Sixty two patients were examined: 10 without mammary disease, 10 with fibrous cystic mastopathy, and 42 with breast cancer, which was hystologically verified. PET scanning was recorded in the "Whole body" mode 70-90 min later administration of 370-420 MBq 18F-FDG. It was shown that PET has a high diagnostic accuracy in breast cancer detection. There were no False-positive cases in our investigations. PET scanning in the "Whole body" mode was allowed to assess dissemination of tumor process with high accuracy. Metastatic involvement of local lymph nodes was detected in 82.8% cases and bone metastases--86% in cases. PMID- 11858030 TI - [Ultrasound study in the diagnosis of colon obstruction]. AB - The authors present the results of their investigations into the use of routine transabdominal sonography and ultrasound irrigoscopy in the diagnosis of acute colon obstruction in 70 patients. In 55 patients, the cause of this pathology was tumor-induced obturation of colon lumen and in 15 cases, the pathology resulted from colon evacuation dysfunction due to congenital abnormalities, acquired stenosis due to inflammatory diseases. Ultrasound irrigoscopy was first applied to determine the presence and degree of colon obstruction and to establish its causes. The procedure and semiotics of ultrasound irrigoscopy and routine transabdominal sonography for this abnormality have been developed and are described in detail. Analysis of the findings has demonstrated that ultrasound irrigoscopy used in the diagnosis of acute colon obstruction is on a par with X ray irrigoscopy in its informative value and may be widely employed in the used arsenal of diagnostic techniques. PMID- 11858031 TI - [Wilm's tumor. Diagnostic capacities of magnetic resonance imaging. MRI pathomorphological comparison]. AB - The accuracy of magnetic resonance imaging (MRI) in the diagnosis of Wilms' tumor (WT) and in the evaluation of preoperative chemotherapy (PCH) efficiency was investigated and compared with histopathological data of 56 children and infants with proven retroperitoneum neoplasma (WT--49, neuroblastoma--6, congenital mesoblastic nephroma--1). The author described the WT MRI-semiotics in general and in particular for its changes during the preoperative chemotherapy. The formula for calculation of tumor reduction index is suggested. The MRI sensitivity (100%), specificity (77.8%) and accuracy (91.1%) are detected. The high positive correlation level between the MRI and pathologic findings, concerning WT dimensions, pseudocapsule presence and safety, tumor structure secondary alterations and tumor spreading was found. At the same time, the specific MRI criteria for the different histological types of WT were not found. MRI is confirmed to be an accurate tool for diagnostic monitoring of patients with WT and other retroperitoneum neoplasms. PMID- 11858033 TI - [X-ray endovascular embolization of the superior rectal artery: New potentialities in the surgical management of chronic hemorrhoids]. AB - Based on the concept of occurrence and development of hemorrhoids from pathologically altered groups of cavernous bodies of the anal submucosa, the author used dearterialization of hyperplastic cavernous tissue for the treatment of hemorrhoids. The treatment involves superselective catheterization and material embolization of the superior rectal artery. The characteristics of 86 remedial embolizations of the superior rectal artery and routine treatments (sclerotherapy, latex-ring ligation, cryotherapy, and surgical hemorrhoidectomy) for chronic hemorrhoids. Emphasis is placed on the advantages of embolization methods over conventional low-invasive and surgical treatment. PMID- 11858032 TI - [Direct coronary arterial stenting without predilatation in patients with coronary heart disease]. AB - The study was undertaken to evaluate the safety and efficiency of direct stenting versus routine stenting with predilation. It included 133 patients. By the decision of operators, direct stenting was conducted in 66 patients (71 stenoses) (Group 1). The remaining 67 patients (73 stenoses) underwent routine stenting with predilation (Group 2). The initial angiographic success of stenting was 100% in Group 1 and 98 in Group 2. Complications were absent. In the direct stenting group, technical problems occurred during a session in 9 (12%) cases. In this group the mean duration of fluoroscopy and the total duration of a session were much less than in the routine stenting group. The mean number of balloons used at dilation per stenosis and the number of dilation sessions per stenosis were much lower in Group 1 than in Group 2. The results of quantitative angiogram analysis before and after a session were similar in both patient groups. Six months following stenting, angiographic restenosis occurred in 7 (10%) patients in Group 1 and in 9 (12%) in Group 2. Direct stenting is a safe and effective treatment for non-occlusive coronary lesions without marked kinks and calcinosis. Direct stenting reduces the duration of fluoroscopy and the total duration of an operation by 50 and 22%, respectively, as compared to predilation stenting. PMID- 11858034 TI - [Diagnosis of left-sided superior cava by echocardiography and magnetic resonance imaging]. PMID- 11858035 TI - [Standardization heart X-ray study]. PMID- 11858036 TI - [Information on the 7th All-Russian Congress of Roentgenologists and Radiologists, Moscow, May 29-June 1, 2001]. PMID- 11858037 TI - [Worth of an animal: agriculture]. PMID- 11858038 TI - [Congenital white muscle disease in a Belgian blue calf]. AB - In this case report about white muscle disease (WMD) in a Belgian Blue herd, the disease is described both as an individual and as a herd problem. Aetiology, diagnosis, and therapy of WMD are discussed. WMD is a disease of animals with muscle damage due to the presence of free radicals. Unsaturated fatty acids in the cell membrane are transformed into a radical form in a chain reaction: a fatty acid next to the fatty acid radical can be transformed into another free radical. In healthy animal the chain reaction is stopped by anti-oxidants such as vitamin E and glutathione peroxidase. WMD can occur when more free radicals are produced than the available anti-oxidants can deal with. The disease occurs in calves, lambs, and foals. PMID- 11858039 TI - [Genetic modification of animals]. AB - Genetic modificated animals are an essential part of modern biotechnology since the years of the seventies of last century. In the past selective crossbreeding of plants and animals took place in order to obtain the desired characteristics but nowadays this is done with the aid of biotechnology i.e. through transgenese, knock-out animals and cloning. A number of fields of application of genetic modification of animals are listed. In order to limit the risks that genetic modificated animals may carry with them the necessary legislation is formulated. A number of risks and the main rules are mentioned. PMID- 11858040 TI - [Botulism in a horse: a concrete possibility in the Netherlands?]. PMID- 11858041 TI - [Foot and mouth disease in (meat)calves: clinical signs and viral shedding]. PMID- 11858043 TI - [Massive destruction of healthy animals is from the Middle Ages]. PMID- 11858042 TI - [Deworming schedule in mixed group of horses and donkeys]. PMID- 11858044 TI - [In a veterinary clinic, all noses should point in the same direction]. PMID- 11858045 TI - [Large breed health care symposium in Venice]. PMID- 11858047 TI - [Dogs on a leash?]. PMID- 11858048 TI - Leveraging change through evaluation and consultation. PMID- 11858050 TI - A report on legislation affecting Texas dentists. PMID- 11858049 TI - New foundation addresses lack of care issues for all Texans. PMID- 11858051 TI - 129th Annual Session report--May 6-9, 1999. PMID- 11858052 TI - Oral and maxillofacial pathology case of the month. Ameloblastoma. PMID- 11858053 TI - The view from Carabelli's Cusp. PMID- 11858054 TI - Nuts and bolts of dental electronic claims. Navigating through the hoopla. PMID- 11858056 TI - Importance of CPR and basic life support. PMID- 11858057 TI - Choice of entity: business organization fundamentals. PMID- 11858058 TI - Dentistry in the new millennium. PMID- 11858059 TI - Acute vs. chronic pain. AB - The differences between acute and chronic pain are many and varied. They are so different from one another that they must be considered separate entities. The chronic pain patient does not fit the traditional acute illness model as conceptualized by patients and healthcare providers. Because of the complex nature of the pain mechanism as a protective "reflex" and the fact that the pain response gets caught up in emotional expression, pain becomes a learned behavior pattern. When the patient who presents to the dental office suffering from pain is found not to respond to conventional methods of treatment, the dentist should first consider the nature of the pain response and the fact that the patient may not meet all the requirements for the acute illness model. The manner in which the patient describes his or her pain can be a major clue as to the temporal classification of the pain, thus allowing the dentist the advantage of better decision-making. Great discernment on the part of the dental practitioner must be exercised in order to provide the optimum care for the patient. It is important for the dentist to consider the fact that there may be no underlying cause for the pain and it may be necessary to make proper referrals for management of this type of patient. At a more practical and human level, patients want to know if their pain will ever completely go away. Patients are frightened that their pain is attributable to some unrecognized pathology (catastrophic thinking). This drives them to search for the ultimate cure. Going from practitioner to practitioner worsens the confusion as the patient hopes that someone will be able to illuminate the problem. By being able to classify the pain into a recognizable and explainable syndrome, the pain practitioner is often able to offer hope to the patient. Although treatment often does not yield a completely pain-free state, understanding the basis for the pain can provide significant relief through proper management. PMID- 11858060 TI - Broad support evident for the emerging specialty of orofacial pain. AB - The emerging field of orofacial pain is being considered by the American Dental Association for full status as a new dental specialty to improve the care for these patients. The broad support among dentists for this initiative stems from an awareness of the benefits the field can provide for dentists and their patients. PMID- 11858061 TI - Orofacial pain mechanism. PMID- 11858062 TI - The prevalence and etiology of temporomandibular disorders and orofacial pain. PMID- 11858063 TI - History and examination of the orofacial pain patient. AB - Taking a history and performing an examination on an orofacial pain patient differs significantly from that in the general dental patient. Orofacial pain dentists must be familiar with the disorders that cause chronic head and neck pain. In addition, they must know the pertinent aspects of history taking with regard to the chief complaint, history of the present illness, the relevance of the past medical and dental history and how the social history can act as an important etiologic and prognostic factor. The clinician must also be versed in the elements of the orofacial pain examination which include evaluation of the TMJ and cervical spine; head, neck and dental examination; and, often, neurological and otolaryngological screening. The clinician should also have a familiarity with blood testing, urinalysis and know the uses and limitations of diagnostic imaging. PMID- 11858064 TI - Migraine. PMID- 11858065 TI - Nonodontogenic toothache. AB - Toothache is a common complaint in the dental office. Most toothaches have their origin in the pulpal tissues or periodontal structures. These odontogenic pains are managed well and predictably by dental therapies. Nonodontogenic toothaches are often difficult to identify and can challenge the diagnostic ability of the clinician. The most important step towards proper management of toothache is to be suspicious that the pain may not be of dental origin. The cardinal warning symptoms of nonodontogenic toothache are as follows (28): a. spontaneous multiple toothaches; b. inadequate local dental cause for the pain; c. stimulating, burning, nonpulsatile toothaches; d. constant, unremitting, nonvariable toothaches; e. persistent, recurrent toothaches; f. local anesthetic blocking of the offending tooth does not eliminate the pain; and g. failure of the toothache to respond to reasonable dental therapy. PMID- 11858066 TI - Oral and maxillofacial pathology case of the month. Odontogenic keratocyst. PMID- 11858067 TI - Drug therapy for the orofacial and TMD pain patient. PMID- 11858068 TI - Bruxism and intraoral orthotics. PMID- 11858070 TI - Pain. PMID- 11858069 TI - Overview of American Heart Association protocols. Part III--P.A.L.S. PMID- 11858072 TI - Mary Kay's ADA annual session notes. PMID- 11858071 TI - Mandibular local anesthesia--a historical perspective. PMID- 11858073 TI - Spit Tobacco Prevention Network (STOPN). PMID- 11858074 TI - Oral and maxillofacial pathology case of the month. Lingual mandibular sequestration and ulceration. PMID- 11858076 TI - Battlefield: mercury--the element, not the planet. PMID- 11858077 TI - [Lung resection in drug resistant tuberculosis patients]. AB - Three and seventy-one case histories of patients operated on for different forms of pulmonary tuberculosis in whom their Mycobacteria tuberculosis (MT) were resistant to bactericidal drugs (BD) were analysed. Fibrocavenous or cavernous pulmonary tuberculosis was detected in most patients (73.3%). Various postoperative complications occurred in 20% of the patients operated on, in them empyema with bronchial fistula was stated in 6.5% of cases. The immediate outcomes of lung resections were satisfactory in 90% of patients and poor (progressive tuberculosis, uncorrected empyema) in 6%, mortality was 4%. Comparing these data with the outcomes of 6033 resections whose efficiency was 96.4% and mortality 1.4% leads to the conclusion that MT resistance to BD negatively affect the outcomes of resections. PMID- 11858078 TI - [Methods for overcoming drug resistance in destructive and progressive forms of pulmonary tuberculosis]. PMID- 11858079 TI - [Prospects for predicting the life expectancy of patients with chronic obstructive lung disease for refining indicators for lung transplantation]. AB - Success of lung transplantation is largely determined by the patients' timely referral for surgery. However, the determinants of survival in patients with chronic obstructive pulmonary disease (COPD), potential lung recipients are to be established. To reveal the determinants of survival in patients with terminal respiratory failure, the data from a follow-up of 59 patients with COPD who met the generally accepted requirements for the recipients of an isolated lung were analyzed. The analysis of some clinical, functional, hemodynamic parameters established that the level of hypoxemia and the value of pulmonary systolic pressure were of the greatest prognostic value. The values of external respiratory function did not affect the patients' survival. Thus, patients with COPD should undergo lung transplantation if they have a pulmonary systolic pressure of over 50 mm Hg, pO2 of under 60 mm Hg, and no benefits from their conservative therapy. PMID- 11858080 TI - [Results of local intervention in the cavern in patients with destructive forms of postoperative relapses of pulmonary tuberculosis]. PMID- 11858081 TI - [Results of surgical intervention depending on duration of preoperative treatment of pulmonary tuberculosis patients]. AB - The case histories of 428 patients operated on for tuberculosis were analyzed. Three groups were identified. They were as follows: 1) 121 patients untreated with bactericidal drugs before surgery; 2) 247 patients treated less than 6 months before it; 3) 160 patients treated more than 6 months before surgery. Various complications due to resection of the lung were observed in 30 (7%) patients undergone surgery. They were 6.6, 6.8, and 7.5% in Groups 1, 2, and 3, respectively. A clinical effect was achieved in 99.8% of cases. The late outcomes of surgical intervention were studied within 1 to 10 years in 354 patients, including 102, 119, and 133 patients in Groups 1, 2, and 3, respectively. Progressive and recurrent tuberculosis was revealed in 7 (6.8%), 11 (9.2%), and 18 (13.5%), respectively. Thus, immediate and late outcomes of surgical treatment were not worse in patients with tuberculomas untreated with bactericidal drugs before surgery than in those who receive long-term therapy that substantially reduces the duration of therapy, which is a most important task of modern phthisiology. So patients should be operated on when they are found to have pulmonary tuberculomas without signs of a progressive tuberculous process. PMID- 11858082 TI - [Use of hemostasis in repeat operations in phthisiopulmonology patients]. AB - Blood coagulative parameters were studied in 14 patients at resurgery for complicated forms of pulmonary tuberculosis. The hypocoagulative syndrome due to changes in all hemostatic links: (vascular thrombocytic, coagulative, and fibrinolytic), was detected in the early postoperative period. These changes compensated on postoperative day 3. On day 7 after surgery the hemostatic parameters returned to the preoperative value. Principles of correction of hemocoagulative disorders are given. PMID- 11858083 TI - [Pathogenetic therapy of asthenovegetative syndrome in pulmonary tuberculosis patients in the postoperative period]. PMID- 11858084 TI - [Immunotherapy of autologous focus of infiltrating lymphocytes in resection of pulmonary tuberculosis]. AB - The development of immunotherapy modalities using active immunologically "trained" cells obtained just from a focus of tuberculous inflammation is a promising direction of the prevention and treatment of recurrent tuberculosis. Thirty patients were examined. A procedure was developed for autoimmunotherapy. The study showed it possible to obtain living, sterile, tuberculous inflammation focus-trophic lymphocytes in the amount sufficient for an immunotherapeutical effect. Autoimmunotherapy was found to reduce the number of postoperative recurrences (6% in the experimental group versus 20% in the control one), promoted prompter recovery of tuberculous bronchitis, earlier normalization of the parameters of systemic and antituberculous immunity after surgery. Densitometric analysis of X-ray films revealed a significant focal consolidation (by 1.37 times) after using focus-infiltrating lymphocytes. Autoimmunotherapy using focus-infiltrating lymphocytes makes it possible to improve the outcomes of surgical treatment of patients with pulmonary tuberculosis by reducing the risk of postoperative recurrent tuberculosis, to apply resection interventions in disseminated and bilateral pulmonary tuberculosis more extensively, to use saving resections aimed at removing the basic focus of a tuberculous lesion if there are disseminated focal seedings, and to promote the solution of a problem of the surgical management of patients with drug-resistant mycobacteria. PMID- 11858085 TI - [Role of surgical methods in treating tuberculosis]. PMID- 11858086 TI - [Immunobiochemical methods in substantiating early operations in patients with cavernous pulmonary tuberculosis]. PMID- 11858087 TI - [Videothoracoscopy in diagnosing and treating exudative pleuritis]. AB - Telethoracoscopy (TT) was performed in 76 patients with exudative pleurisy of unclear etiology. A correct diagnosis was made in all (100%) cases. Tuberculosis was detected in 41 (54%) patients. In this group of patients, the results were analyzed by taking into account the duration of disease, the nature of an operation, and morphological findings. In tuberculous pleurisy, TT may be conducted at any time; however, it is most effective when an acute period subsides and an exudate begins forming. At this time (2-3 months after the onset of the disease), surgery is not diagnostic, but also remedial as sanitation of the pleural cavity with partial pleurectomy. PMID- 11858088 TI - [Pleurectomy and decortication in chronic tuberculous pleuritis and pleural empyema]. AB - The study covered 105 patients with chronic tuberculous pleurisy and empyema who had undergone surgery. Reparative pleurectomies with lung decortication were made in all the patients. Four groups of patients operated on in different periods were analyzed. The paper presents clinical, morphological, and instrumental evidence for indications for surgery. The best outcomes were achieved in patients with a 3-4-month history (100% efficiency with recovered ventilation function). The total efficiency of pleurectomy and decortication was 92.4% without fatal outcomes. PMID- 11858089 TI - [Clinical course of tuberculous pleuritis in extreme northern conditions]. PMID- 11858091 TI - [Surgery on osteoarticular tuberculosis in children]. PMID- 11858090 TI - [Atlantoaxial dislocation in spondylitis]. AB - The paper summarizes the outcomes of surgical treatment in 6 patients with nonspecific and tuberculous spondylitis of the craniovertebral area. These patients underwent decompressively stabilizing operations that consists of two steps: at first occipitospondylodesis was performed with a wire and protacryl then via transpharyngeal access with sanitation of the abscess cavity, by removing necrotic tissues--a saving resection of bony tissue sites within the healthy tissues, anterior stabilization of an affected part with an osseous autograft. The above procedure of surgical treatment along with bactericidal therapy yielded positive results by recovering spinal cord function, by forming a bony unit at the site of spinal inflammatory lesion. PMID- 11858092 TI - [Results of decompression operations in patients with tuberculous spondylitis]. AB - The paper summarizes the outcomes of decompression and decompression-reparative operations in patients with tuberculous spondylitis. Good surgical results with complete elimination or alleviation of neurological disorders have been achieved in 94.8% of cases. PMID- 11858093 TI - [Features of surgery on female genital tuberculosis]. AB - The paper gives the outcomes of treatment in 4983 patients treated at the Gynecology Clinic of the Saint Petersburg Research Institute of Phthisiopulmonology in 1977 to 1999. Surgical interventions different in their scope were made in 949 (19%) females. Analyzing 526 females of them indicated that organ-saving operations were performed in most cases (80.2%). Indications for surgery and the time of preoperative etiotropic therapy were defined. Emphasis is laid on that the introduction of laparoscopy into practical phthiosiologic gynecology will solve many problems associated not only with a surgical appliance, but also with the diagnosis of genital tuberculosis. PMID- 11858094 TI - [Genital tuberculosis and surgical treatment of it]. AB - The paper presents the specific features of diagnosis and clinical course of genital tuberculosis under the present conditions. It shows it expedient to make a differential diagnosis of genital tuberculosis in 2 steps: 1) the use of the routine examination and 2) the study of specific antituberculous immunity. Genital tuberculosis risk groups to be obligatorily examined at a specialized tuberculosis control facility are identified. The specific features of the clinical course of genital tuberculosis, such as aptness to exudative processes, a combination with extragenital tuberculosis and hormonally depended gynecological diseases are noted. Indications for and efficiency of endosurgical interventions in genital tuberculosis are shown. PMID- 11858096 TI - [Surgical treatment of patients with abdominal tuberculosis]. PMID- 11858095 TI - [Surgery on tuberculosis lymphatic system and abdominal organs]. AB - The paper presents data on surgery for tuberculosis of the lymphatics and abdominal organs. Removal of tuberculosis-affected peripheral lymph nodes during etiotropic therapy yields positive therapeutical results in 84.6% of patients. The efficiency of early operations has been first evidenced by a dynamic study of the systemic immunity. Complicated abdominal tuberculosis is encountered in 44.2% and forced emergency and planned operations on the abdomen to be performed; good late results should be consolidated by combined bactericidal therapy. PMID- 11858097 TI - [A modern look at the problem of spontaneous pneumothorax]. PMID- 11858098 TI - [Significance of mycobacterial drug resistance in pulmonary tuberculosis surgery]. AB - The outcomes of surgical treatment of 546 patients with pulmonary tuberculosis were analyzed in relation to the bacteriological characteristics. Seeding detected sputum bacterial isolation in 52.4% of cases. Mycobacterial drug resistance was detected in 83.6% of the tested cultures. It has been concluded that the resistance is a first-order infectious agent whose action is shown in lowering the efficiency of preoperative courses of chemotherapy by 2.5 times, in increasing the incidence and severity of postoperative pleural and pulmonary complications by 6-7 times, in deteriorating the immediate outcomes of surgical treatment to 82.4% with a 7.4% mortality rate. Intensive bacterial isolation unarrested by surgery is a second-order infectious agent which more clearly shows a relationship of the efficiency of surgical treatment to a reduction in the cure rate to 74.2% with a total mortality of 15.2%. There is evidence for that it is advisable to apply an active surgical policy in a group of patients having an infectious risk factor. PMID- 11858099 TI - [A natural course of untreated hyperprolactinemia]. PMID- 11858100 TI - [Levels of anti-inflammatory cytokines interleukin-2, interleukin-8, and soluble interleukin-2 receptor in blood of patients with various forms of ischemic heart disease]. AB - AIM: To quantify interleukin-8 (IL-8), interleukin-2 (IL-2) and soluble receptor of IL-2 (sIL-2r) in blood serum of patients with various forms of ischemic heart disease (IHD). MATERIAL AND METHODS: Levels of IL-8, IL-2 and sIL-2r were measured with enzyme immunoassay (EIA) in the serum of 75 patients with IHD: angina of effort (group 1), progressive angina (group 2) and acute myocardial infarction (group 3). The EIAs were performed at admission and 2 weeks later. RESULTS: Baseline levels of IL-2 in group 1 and 2 patients were close (9.1 +/- 1.6 and 10.1 +/- 3.8 pg/ml) being significantly lower in group 3 (0.81 +/- 0.57 pg/ml, p < 0.01). 14 days of therapy did not change the values noticeably. IL-8 level was the highest in group 1 (94.2 +/- 27.6, 20.03 +/- 7.4, 22.47 +/- 4.8 pg/ml, respectively). sIL-2r in the three groups did not vary greatly (73.95 +/- 12.23, 89.46 +/- 18.17, 89.2 +/- 14.17 pg/ml, respectively). SIL-2r levels rose in 2 weeks in group 3 (to 147.67 +/- 18.17 pg/ml). CONCLUSION: It is confirmed that IL-2, IL-8 and sIL-2r take part in pathogenesis of IHD. IL-2 and IL-8 levels are persistently high in anginal patients while in patients with acute myocardial infarction they are low. Low concentrations of IL-2 in the latter may be attributed to high levels of its soluble receptor. PMID- 11858101 TI - [Changes in lipid metabolism in platelet membranes in patients with ischemic heart disease following meals with animal and vegetable fats]. AB - AIM: To evaluate changes in oxidative lipid metabolism in platelet membranes by content of lipid peroxidation (LPO) products and antioxidant defense (AOD) activity in patients with stable chronic ischemic heart disease (IHD) after single intake of animal fat or vegetable oil. MATERIAL AND METHODS: 30 IHD patients with stable angina of effort functional class II-III and 30 healthy controls had a single meal with animal fat or vegetable oil. Before the meal and 6 hours after it tests were performed for LPO activity and AOD in platelet membranes. RESULTS: Patients with stable chronic IHD showed LPO activation and suppressed enzymatic and non-enzymatic AOD. Single animal fat or vegetable oil intake cause insignificant shifts in oxidative platelet metabolism in healthy controls while animal fat in IHD patients activates LPO and promotes accumulation of initial and end peroxidation products in platelet membranes. Vegetable fat in IHD patients suppresses enzyme AOD. CONCLUSION: Animal fat and vegetable oil as a single diet load have a prooxidant effect at the levels of platelet membranes in IHD patients but not in healthy persons. PMID- 11858102 TI - [Tolerance to nitrates in patients with angina and effect of plasmapheresis]. AB - AIM: To evaluate tolerance to isosorbide dinitrate (ID) in patients with angina pectoris functional class II-III and effect of plasmapheresis in this tolerance management. MATERIAL AND METHODS: 85 patients with angina pectoris functional class II-III taking long-acting ID. RESULTS: As shown by repeat transesophageal left atrial pacing, 24.7% patients have developed tolerance to nitrate while taking prolonged ID. Two methods of this tolerance elimination were compared: plasmapheresis + withdrawal of long-acting nitrate and withdrawal of the nitrates only. The former has shown some advantages: use of plasmapheresis reestablished ID efficiency in 4 days (two sessions) in 81.8% and in 7 days (3 sessions) in 100% patients without withdrawal syndrome. The latter method reestablished a marked antiischemic effect of ID in 7 days and 20% patients exhibited a variant of withdrawal syndrome--early negative aftereffect. CONCLUSION: When withdrawal of long-acting nitrates is combined with plasmapheresis in anginal patients to eliminate tolerance to nitrates, the effect is better. PMID- 11858103 TI - [Comparative efficacy of tiklid and aspirin in patients with unstable angina]. AB - AIM: To evaluate effects of tiklid and aspirin on hemostasis, antithrombogenic activity of vascular wall in patients with unstable angina (UA). MATERIAL AND METHODS: An open randomized trial enrolled 30 UA patients given tiklid (n = 16) or aspirin (n = 14). Hemostasis, platelet aggregation, antithrombogenic activity of the vascular wall were examined on the treatment day 1 and 20. The response was assessed by the number of anginal attacks, dose of nitroglycerine, data of Holter monitoring. RESULTS: Tiklid was more effective than aspirin in UA patients as it by the treatment day 20 lowered platelet aggregation more, higher elevated anticoagulation potential of the blood and vessels, and stronger activated fibrinolytic hemostasis. CONCLUSION: Compared to aspirin, tiklid is more potent antiaggregant, has greater effect on antithrombogenic properties of the vascular wall. PMID- 11858104 TI - [Melatonin production in hypertonic patients during magnetic storms]. AB - AIM: To study mechanisms of action of natural magnetic field of the Earth on arterial pressure (AP) and melatonin production in patients with essential hypertension (EH) stage II. MATERIAL AND METHODS: Clinical, laboratory and device investigations covered 52 men with EH stage II (mean age 42 +/- 0.92 years) and 11 healthy men (mean age 23 +/- 1.46 years). Mean 24-hour, mean daytime, mean night systolic and diastolic pressures, 24-h index, time hypertensive index, standard deviation were registered. Melatonin was measured in the urine by radioimmunoassay. Geomagnetic situation was assessed by K-index (quiet--under 15, disturbed--15-25, magnetic storm--above 25). RESULTS: In hypertensive patients AP grew with growth of geomagnetic activity. In normal subjects AP remained normal. The 24-h rhythm of AP variability in hypertensives was normal. Magnetic storm affected melatonin production in EH patients noticeably: night and daytime production of melatonin was low. In normal subjects night melatonin production was high. CONCLUSION: AH stage II patients respond to magnetic storm with maladaptation, i.e. a rise in AP and low melatonin production. PMID- 11858105 TI - [The novel method for selection of preparations for treatment of arterial hypertension based on spectral analysis of heart rhythm variability]. AB - AIM: To study spectral characteristics of heart rhythm in patients with arterial hypertension (AH) to select medication for hypertension and control over the treatment efficacy. MATERIAL AND METHODS: 35 AH patients were examined with general clinical methods and using spectral analysis of variability of heart rhythm (VHR). Five spectrograms were registered. Each of them contained three peaks: very low, low and high frequencies (VLF, LF and HF, respectively). Normal values of VLF, LF and HF were estimated on the basis of spectral values obtained in healthy subjects. After registration of spectral values all the patients were given at first placebo, than enalapril, one day after its discontinuation- mifedipine. At each stage VHR was recorded. Changes in the spectral indices were compared to changes in clinical symptoms. RESULTS: VLF proved most important for a rapid choice of adequate antihypertensive therapy. CONCLUSION: Normalization of VLF, LF and HF indicates adequacy of the choice of antihypertensive treatment. PMID- 11858106 TI - [Sinus node functions, sinoatrial conduction, atrial conductivity after incipient paroxysms of atrial fibrillation and flutter in patients with ischemic heart disease]. AB - AIM: Prediction of the rate of recurrent paroxysms of atrial fibrillation (AF) and flutter (AFl) after the first arrhythmia episode; determination of relevant antiarrhythmic treatment. MATERIAL AND METHODS: 157 patients with ischemic heart disease (IHD) complicated by new episodes of AF and AFl entered the study. After the initial episode and 1-2 arrhythmia recurrences all the patients have undergone assessment of hemodynamics, atrial conduction of excitation, sinus node function using transesophageal pacing. The patients were divided into two groups: group 1 consisted of 42 patients having no recurrent paroxysms of AF or AFl for at least 6 months; 115 patients of group 2 had at least one episode of recurrent arrhythmia for 6 months after the first paroxysm. RESULTS: Patients of group 2 vs those of group 1 had a significantly longer first episode, more frequent occurrence of calcinosis of mitral and/or aortic valve, more serious systolic and diastolic dysfunctions and most frequent retrograde atrial excitation conduction after the first paroxysm. CONCLUSION: In detection of only disturbed intraatrial conduction in IHD patients after the first paroxysm of AF and AFl predicted are clinical recurrences of arrhythmia with the recurrence-free period more than 6 months. In retrograde atrial conduction of excitation combined with systolic and diastolic left ventricular dysfunction, sinus node dysfunction prognosis was made of more frequent episodes of AF and AFl. PMID- 11858107 TI - [Characteristics of arrhythmia and late ventricular potentials in patients with cardiological syndrome X]. AB - AIM: A comparative study of late ventricular potentials (LVP) in patients with cardiological syndrome X (SX) and stenotic atherosclerosis of coronary arteries (ACA) as well as their relations with arrhythmia, cardiac contractility, lipid metabolism and morphological characteristics of the myocardium. MATERIAL AND METHODS: The examination of 52 SX and 77 ACA patients as well as 17 healthy subjects included coronaroventriculography, bicycle exercise, 24-h ECG monitoring, echocardiography, signal-averaged high-resolution (SAHR) ECG, investigation of blood lipoproteins. Endomyocardial biopsy was made in 5 ACA and 5 SX patients. RESULTS: No differences were registered between SX and ACA patients by frequency and severity of arrhythmic episodes, percentage of patients with registered LVP, quantitative parameters of SAHR ECG. Frequency of high gradation ventricular arrhythmia episodes was significantly higher in SX and ACA patients with LVP than such patients free of LVP. SX patients had correlation between parameters of SAHR ECG, myocardial contractility and lipid metabolism. Foci of diffuse cardiosclerosis are most probable anatomic substrate of LVP. CONCLUSION: The risk of prognostically unfavourable high-gradation ventricular arrhythmia episodes in SX and ACA patients is the same. LVP may predict severe ventricular arrhythmia episodes both in SX and ACA patients. PMID- 11858108 TI - [Autonomic nervous system balance, variability and arrhythmia in survivors of myocardial infarction]. AB - AIM: To investigate relations between heart rhythm variability (HRV), vegetative balance and electric myocardial activity in myocardial infarction (MI) survivors. MATERIAL AND METHODS: HRV was studied by short 5-min parts of ECG and data of ECG Holter monitoring were analysed for 98 patients who had macrofocal MI 1.5 months to 5 years before. RESULTS: Manifestations of electric heart instability were polymorphic. 69.4% examinees had hyperactivity of the parasympathetic nervous system (PSNS). The influence of the sympathetic nervous system (SNS) increased with growing severity of arrhythmia. Supraventricular arrhythmia occurred more frequently in high PSNS activity, while ventricular arrhythmia occurred more often in SNS prevalence combined with low HRV. CONCLUSION: HRV analysis for MI survivors, especially in combination with Holter ECG monitoring, gives an objective assessment of various manifestations of cardiac dysfunction and therefore enables timely adequate therapy. PMID- 11858109 TI - [Obesity and insulin resistance--risk factors and parts of metabolic syndrome]. PMID- 11858110 TI - [Structural and electrophysiological characteristics of the heart function during paroxysmal atrial fibrillation]. AB - AIM: To examine structural and electrophysiological predictors of the onset and establishment as chronic of cardiac fibrillation (CF). MATERIAL AND METHODS: Echocardiography, transesophageal electrophysiological examination of the heart were performed in 420 cases of paroxysmal cardiac fibrillation (PCF). They were diagnosed as having new-onset (n = 68), rare (n = 92) and frequent (n = 260) PCF. RESULTS: Compared to healthy subjects, PCE patients had larger size of the left atrium (LA), higher atrium/ventricle value, lower frequency threshold of arrhythmia induction, decreased left atrial effective refractory period, longer induced paroxysm and maximal atrial response. CF chronicity can be predicted with the frequency threshold of arrhythmia induction under 300 imp/min, left atrial effective refractory period under 220 ms, left atrial size over 42 mm, induction of persistent PCF. CONCLUSION: A significant correlation was found between left atrial size and electrophysiological parameters of the heart characterizing arrhythmogenic "readiness" of the atria. This suggests a provoking role of left atrial dilatation in PCF. PMID- 11858111 TI - [Respiratory sleep disorders in patients with congestive heart failure]. AB - AIM: To examine respiration in sleeping patients with congestive heart failure (CHF). MATERIAL AND METHODS: Night cardiorespiratory monitoring was made in 30 CHF patients with coronary heart disease (CHD), arterial hypertension and dilated cardiomyopathy. The control group consisted of 16 patients with obstructive sleep apnea and hypopnea (OSAH) free of CHF. RESULTS: Respiratory sleep disorders were registered in all the examinees. Moderate and severe OSAH occurred more often. CHF patients vs controls had shorter inhalation, expiration, overall duration of the respiratory cycle, higher rate of the respiratory movements. With aggravation of the respiratory disorders in sleeping CHF patients, minute ventilation, respiratory volume and mean inspiratory flow arose. In severe OSAH there was low functional residual lung capacity and high intraalveolar volume in end expiration positive pressure. The above changes may cause fatigue and weakness of the respiratory muscles and alter intracardiac hemodynamics. CONCLUSION: For CHF patients it is recommended to perform CPAP-therapy in sleep. PMID- 11858112 TI - [Fraxiparin in the treatment of thrombophilia in angiopathy]. PMID- 11858114 TI - [Post-respiratory infection reactive arthritis--etiology, clinical and laboratory characteristics and early prognosis]. AB - AIM: To study etiology, clinico-laboratory characteristics and early prognosis of reactive arthritis developing after respiratory infections (priReA). MATERIAL AND METHODS: Blood samples from 69 priReA patients aged 17-45 years were examined for specific antibodies to 28 bacterial and viral infectious agents, and antigens of group HLA-B7-CREG (B7, B13, B22, B27, B40, B73, Cw2). The test were made in hospital and 3 months after the patients' discharge. RESULTS: The priReA respiratory etiology was discovered in 31 patients, urogenital--in 14 cases and enterocolitic one--in 7 patients. Streptococcus and pneumoneal chlamydia cause priReA most frequently. HLA-B40 carriers are more predisposed to priReA. priReA runs with less frequent Reiter's syndrome and lesions of axial and extraarticular structures, better early prognosis. CONCLUSION: In young patients with early ReA respiratory etiology is observed more frequently than urogenital and enterocolitic ones. Compared to other kinds of ReA, priReA demonstrates some genetic, clinical and prognostic peculiarities. PMID- 11858113 TI - [Quality of life and characteristics of the external respiration function in patients with bronchial asthma]. AB - AIM: To investigate correlations between quality of life, bronchial obstruction, nonspecific bronchial hyperreactivity (NSBH) and response of these parameters to treatment with inhalation steroids in patients with bronchial asthma (BA). MATERIAL AND METHODS: The study included 67 BA patients (47 women and 20 men) aged 18 to 62 years (mean age 43 years). 59 of them had the diagnosis of atopic BA, 8 had bacterial BA. Quality of life was assessed with the questionnaire AQLQ, bronchial obstruction was judged by forced expiratory volume per a second (FEV1). Bronchial hyperreactivity was studied with acetylcholine and histamine tests. RESULTS: Quality of life strongly correlated with FEV1 and NSBH. Positive changes in the latter due to treatment with inhalation glucocorticoid budesonide (400 mcg/day) entailed improvement of life quality. CONCLUSION: Bronchial obstruction and NSBH have a significant effect on BA patients' life quality which should be considered in monitoring of BA patients' condition. PMID- 11858115 TI - [Bone mineral density in patients with rheumatoid arthritis]. AB - AIM: To examine the prevalence of osteoporosis (OPO) and osteopenia (OPE) in female patients with rheumatic arthritis (RA). MATERIAL AND METHODS: 60 female patients with proved diagnosis of RA aged 34-64 years: 30 premenopausal women (median age 41.5 years, disease duration 9.5 years) and 30 postmenopausal women (median age 56.2 years, disease duration 10.2 years). Both groups have not undergone any glucocorticoid or antiosteoporotic therapy. Bone mineral density (BMD) was measured with a Dual Energy X-Ray Absorptiometry (DEXA) in the lumbar spine, proximal segments of the femur, forearm. RESULTS: In the group of premenopausal patients with RA the rate of OPE was 63% for forearm, 60% for femoral neck, 33% for lumbar spine. In postmenopausal women 53, 50 and 50%, respectively. CONCLUSION: Osteoporosis in RA is of a generalized character and can be encountered in peripheral skeleton more often than in the axial one. It is possible to estimate the BMD of one section by means of the BMD indicator of another section. PMID- 11858116 TI - [Intake of non-steroid preparation and risk of developing cancer of the upper gastrointestinal tract]. AB - AIM: To compare occurrence of upper gastrointestinal tract (UGT) cancer in rheumatic patients taking long courses of nonsteroid antiinflammatory drugs (NSAD) and in non-rheumatic patients who have not received NSAD. MATERIAL AND METHODS: The study group of 1271 rheumatic patients on NSAD for at least 12 months. Control group of 654 patients free of rheumatic diseases and having no history of regular NSAD intake. The groups did not differ by the mean age, erosion and ulcer incidence. RESULTS: Esophagogastroduodenoscopy detected and histology confirmed adenocarcinoma in 2 patients of the study and 10 patients of the control group (0.16 and 1.5%, respectively, p < 0.001). CONCLUSION: Long-term treatment with NSAD lowers the risk of UGT tumors. PMID- 11858117 TI - [Various aspects of bone marrow cell proliferation in acute leukemia]. AB - AIM: To study proliferation of bone marrow cells in acute leukemia using flow DNA cytometry. MATERIAL AND METHODS: Parameters of the cell cycle (S, G2 + M, G1/0) and myelogram were studied in 55 patients with acute myeloid leukemia (AML) and 46 patients with acute lymphoblastic leukemia (ALL) in remission (135 cases) and acute period (66 cases). Only cases with cell clones containing normal (diploid) number of DNA were included. RESULTS: In acute period, distribution of AML cases by the percentage of cells in S-phase of the cellular cycle was normal, in ALL- log-normal. In remission it was lognormal in both leukemias. Variability of cell cycle parameters in leukemia in some patients was much less than a mean sample one. The reestablishment of proliferation after chemotherapy (7 + 3) in AML was followed up. CONCLUSION: If proliferative activity of blast (tumor) cells in AML is subnormal (10%), proliferation of myelokaryocytes can be used as an additional test for assessment of remission completeness. PMID- 11858118 TI - [Major aspects of pathogenesis in atherosclerosis: current status and perspectives]. PMID- 11858119 TI - [The role of cytokine system in pathogenesis of chronic cardiac insufficiency]. PMID- 11858120 TI - [The role of neutrophils in ischemic and reperfusion myocardial injury]. PMID- 11858121 TI - [Clinical and morphological characteristics of kidney pathology in antiphospholipid syndrome]. PMID- 11858122 TI - [Myocardial and vascular impairment in diabetes mellitus]. AB - AIM: To study relations between ischemic heart disease (IHD) and hormonal and metabolic disorders in patients with diabetes mellitus and IHD. MATERIAL AND METHODS: Cuff test was used to study functional activity of the vessels. Platelet aggregation, prostacyclin, antithrombin III were studied in 168 healthy subjects, diabetes mellitus (DM) patients (n = 98) and DM type 2 patients (n = 118). RESULTS: Microscopic examinations of the myocardium (endomyocardial biopsy) were made and the results were compared to antithrombogenic activity of vascular wall in DM. Myocardial vessels in the patients have undergone morphological alterations. Antithrombogenic activity of the vessels in DM patients was studied under compression. CONCLUSION: Microcirculation was found changed: vascular walls were thick and homogenized, endothelyocytes underwent edema and destruction with enlargement of the basal membrane. Platelet hypersensitivity was proved in DM as well as more active metabolism of arachidonic acid, low prostacyclin endothelial activity. PMID- 11858123 TI - [Apoptosis and inflammation in bronchial asthma]. PMID- 11858124 TI - [Chronic myeloid leukemia: current pathogenetic aspects and new therapeutic approaches (a foreign literature review)]. PMID- 11858125 TI - Keynote address at the opening ceremony for the centennial of the Pan American Health Organization. PMID- 11858126 TI - Obstetric complications: does training traditional birth attendants make a difference? AB - OBJECTIVE: To assess the effect that a training intervention for traditional birth attendants (TBAs) in Guatemala had on the detection of obstetric complications, the referral of patients with complications to the formal health care system, and, ultimately, those patients' utilization of essential obstetric care services. METHODS: Using a quasi-experimental design, a surveillance system of births was implemented to collect population-based information from 3,518 women between 1990 and 1993. All women were interviewed postpartum by physicians. There were three key independent variables in our study: 1) geographical area (intervention community and non-intervention community), 2) time in relation to the training intervention (before or after), and 3) presence or absence of a TBA at the time of the complication. The key dependent variables for women interviewed were 1) development of an obstetric complication, 2) detection of the problem by the TBA, 3) referral to a health facility, 4) compliance with referral, and 5) use of services. RESULTS: The incidence of postpartum complications decreased after the intervention, controlling for intervention community. On the other hand, after the intervention TBAs were less likely to recognize most maternal complications, and referral rates did not increase significantly. The likelihood of using health care services increased six-fold among women who were not attended by TBAs, and no increase was observed among those who were attended by TBAs. CONCLUSION: Training TBAs may have had a positive effect on the rate, detection, and referral of postpartum complications. However, the evidence is less convincing for overall increases in the detection of complications, in referral to the formal health care system, and in the utilization of essential obstetric services among women attended by TBAs. PMID- 11858127 TI - [Impact of the worker food program in Brazil]. AB - OBJECTIVE: The worker food program of the Brazilian Ministry of Labor and Employment attempts to ensure adequate nourishment for low-income workers. This study evaluates the nutritional impact of the program, which currently serves approximately 10 million workers. METHODS: This retrospective dynamic cohort study was carried out with 8,454 workers living in the state of Bahia, Brazil. Health-related data from January 1996 through April 2000 were obtained from computerized medical records from an organization that provides health surveillance services to a large number of companies in the state. The analysis utilized demographic data and nutritional data (weight and also preobesity, that is, a body mass index of 25.00-29.99). Additional information was obtained from the companies through telephone interviews. RESULTS: Being covered by the food program was positively associated with weight gain (age- and sex-adjusted incidence-density ratio = 2.21; 95% confidence interval: 1.78-2.75). No statistically significant association was found between the program and preobesity. The risk of weight gain was higher (P < 0.05) among workers who, at baseline, were of normal weight (body mass index of 18.50-24.99), were preobese, or were from a low socioeconomic background. CONCLUSIONS: These results suggest that the worker food program has an unintended negative impact on the nutritional status of workers from a low socioeconomic background. The program's approach, which is limited to dietary recommendations concerning the caloric content of meals, should be revised to better promote the health of the workers participating in the program. PMID- 11858128 TI - [Malaria in children: links between nutrition and immunity]. AB - OBJECTIVE: To explore some relationships among immunity, nutrition, and malaria in a group of children from a population with a high endemic incidence of malaria. METHODS: This pilot descriptive prospective cross-sectional study was carried out in 1998 in the municipality of El Bagre, Antioquia, Colombia. Both boys and girls were studied in two groups: one group of 51 children had malaria and another group of 49 did not have malaria. The children with malaria had symptoms or signs of malaria and a positive thick blood film. The comparison group of 49 children without malaria (no signs or symptoms of malaria and a negative thick blood film) were chosen at random at government schools or children's centers. For the study the following indices were calculated: weight for-age, height-for-age, and weight-for-height. Also measured were the blood serum concentrations of: albumin, prealbumin, apolipoprotein A1 (apoA1), transferrin, zinc, vitamin A, immunoglobulins G and M, interleukin-10 (IL-10), tumor necrosis factor-alpha, interferon-gamma, and lymphocyte populations. RESULTS: Of the children studied, 69% presented some risk of malnutrition, and 63% had some risk of chronic malnutrition. With regard to the immunity and biochemical variables, the children with malaria had less apoA1 and albumin and more IL-10 than did the children without malaria. All the biochemical variables showed lower averages in the group with malaria, risk of malnutrition, and immune or biochemical changes, while all the immunity variables had higher averages in that same group of children. CONCLUSIONS: 1) The high frequency of chronic malnutrition found clearly indicates the need for food supplementation measures. 2) The low values of prealbumin found could be due to the frequent bacterial or viral infections reported. 3) The serious vitamin A deficiency found calls for a supplementation program. 4) An association was found between low apoA1 values and the presence of malaria, but which one follows from the other is not known. 5) No relationship was observed between the anthropometric indicators of risk of malnutrition and the possible biochemical markers of malnutrition. 6) We found high levels of IL-10 in the children with malaria; this is the first time that this has been reported for Plasmodium vivax. PMID- 11858129 TI - [Emerging bacterial zoonoses]. AB - Different zoonoses whose etiologic agents were previously unknown have been identified in recent years as a result of new diagnostic techniques in molecular biology and improved epidemiological surveillance systems. Several factors have facilitated the emergence of new diseases and the reemergence of already known diseases, which were thought to be under control. They include the following: (1) environmental changes, such as deforestation and droughts, which allow etiologic agents and their vectors to thrive, (2) industrial food production for widespread consumption, which allows disease agents to spread quickly, (3) certain risky human habits, especially among immuno-compromised persons, such as having exotic pets at home, and (4) the adaptation of etiologic agents to new environmental conditions through the development of drug resistance. Other factors, such as the disruption of established health systems during the process of privatization, have also contributed to the situation. In most cases, bacterial zoonoses can be prevented through good personal and domestic hygiene and common sense. This report is not an exhaustive presentation of all emerging and reemerging bacterial zoonoses. Rather, it aims to illustrate, through numerous examples, the various factors that have contributed to the epidemiological changes that have taken place since the latter part of the twentieth century. PMID- 11858130 TI - Language barriers contribute to health care disparities for Latinos in the United States of America. PMID- 11858131 TI - The scoop on baby's poop. PMID- 11858132 TI - Introduction: linkage analyses in the Hutterites. PMID- 11858133 TI - Introduction: interactions in asthma.. PMID- 11858134 TI - Introduction: transmission/disequilibrium test/association group. . AB - A range of study designs, using unrelated or family controls, were used to investigate the pattern of association with disease of single nucleotide polymorphisms (SNPs) within candidate gene 1 (simulated data). Strong evidence of disease association at the functional locus was detected using all study designs, and in the "general" but not the "isolated" population the functional polymorphism displayed considerably higher association than surrounding SNPs. There was much variation in the strength of association of SNPs with disease, up to 70% of which was explained by SNP allele frequency and distance from the functional polymorphism. Some common polymorphisms very close to the functional locus however showed no association with disease. Analysis of short haplotypes of SNPs reduced but did not totally remove this feature. PMID- 11858135 TI - Introduction: linkage analysis of quantitative traits. PMID- 11858136 TI - Genome-wide linkage analysis in a general population sample using sigma 2A random effects (SSARs) fitted by Gibbs sampling. AB - We used variance components analysis to investigate the underlying determinants of the quantitative phenotypes (Q1-Q5) and their interrelationships in replicate 42 of the Genetic Analysis Workshop 12 simulated general population. Variance components models were fitted using Gibbs sampling in WinBUGS v1.3. Sigma-squared A-random-effects (SSARs) were estimated for each phenotype, and were used as derived phenotypes in subsequent linkage analyses. Whole-genome, multipoint linkage analyses were based upon a new Haseman-Elston identity-by descent sib pair method that takes a weighted combination of the trait-sum and trait difference. The five quantitative traits simulated were closely correlated with each other and with affection status. The whole-genome screen of quantitative traits associated with the simulated complex disease suggested that one or more major loci regulating Q1 localizes to chromosome 2p and that one or more major loci regulating Q5 may localize to chromosome 1p. PMID- 11858137 TI - Setting case management caseloads remains tricky business. AHC/CMSA caseload survey. PMID- 11858138 TI - Athens Declaration on Mental Health and Man-made Disasters, Stigma and Community Care. PMID- 11858139 TI - Healthy selfishness--me first, then you. PMID- 11858141 TI - Healthy Aging for Functional Longevity: Molecular and Cellular Interactions in Senescence. Proceedings of a conference. February 2000, Kyongju, South Korea. PMID- 11858140 TI - Embrace life: fulfill your life purpose. PMID- 11858142 TI - Catheter-based myocardial gene transfer for therapeutic angiogenesis in patient with chronic myocardial ischemia. PMID- 11858144 TI - Progressive non-infectious anterior vertebral fusion ('Copenhagen disease'). PMID- 11858145 TI - The National Task Force on Hepatitis B Immunization, Focus on Asians and Pacific Islanders: a chronicle of achievement. AB - PURPOSE: The goal of The National Task Force on Hepatitis B Immunization, Focus on Asians and Pacific Islanders (TF) is to achieve a 90% hepatitis B vaccination rate of children born before 1993 by the Year 2004. The purpose of this article is to provide a historical framework and a discussion of the achievements by the TF on the efforts taken to achieve its goal. METHODS: A review of all historical TF records, including TF minutes, quarterly reports, annual reports, newsletter articles, journal articles and other documents and Web sites was undertaken. PRINCIPAL FINDINGS: The TF has achieved and exceeded many of its objectives, and has been particularly effective in its public policy and public education endeavors. It has been a national catalyst for organizing hepatitis B catch-up efforts for those children who have not received a full 3 dose complement of hepatitis B vaccine. CONCLUSIONS: It takes the dedicated work of many individuals and organizations to achieve a goal as ambitious as that of the TF. However, the TF has garnered significant volunteer support. The efforts of these dedicated individuals must be recognized. It must also be recognized that without significant financial backing and technical support, the work of these individuals is limited. The TF demonstrates many of the hallmarks of a successful coalition, and through its leadership, will continue to strive toward its goal. RELEVANCE: This article is relevant in that it outlines the significant achievements of a dedicated group of individuals to solve a serious public health problem, that of eliminating hepatitis B as a risk for our children. It describes the TF achievements, but also points to the barriers that a coalition can face in the light of limited resources and the demands of time. This article describes the elements of an effective coalition and compares the work of the TF to those elements. PMID- 11858146 TI - [Superiority of interferon beta-1b in comparison with interferon beta-1a in multiple sclerosis. 2 years outcomes reveal that clinical advantages increase over time]. PMID- 11858147 TI - [How do immune cells react to pathogen invasion?]. PMID- 11858148 TI - Quadricoccus australiensis gen. nov., sp. nov., a beta-proteobacterium from activated sludge biomass. AB - A gram-negative coccus, designated strain Ben 117T, was obtained in axenic culture by micromanipulation from an Australian activated sludge biomass sample, which had been subjected to chlorination in order to alleviate problems associated with foaming and bulking. This isolate was a strict aerobe and grew in axenic culture, also appearing in biomass samples as cocci or clusters of cocci in tetrads, thus resembling the morphotype 'G-bacteria' seen commonly in activated sludge samples. Strain Ben 117T was non-motile, aerobic, oxidase negative and catalase-positive and grew between 15 and 30 degrees C, with an optimum of 25-30 degrees C. The pH range for growth was between 6.0 and 8.5, with an optimum of 7.5-8.5. The isolate stained positively for intracellular polyphosphate and poly-beta-hydroxybutyrate and its G+C content was 67 mol%. 16S rDNA sequence analysis suggests that strain Ben 117T is phylogenetically different from members of the genera Amaricoccus, gram-negative 'G-bacteria' isolated previously in this laboratory. Ben 117T is a member of the Rhodocyclus group in the beta-Proteobacteria and equidistantly placed (similarity value of 95%) between Ferribacterium limneticum and Dechloromonas agitata (mean similarity value of 92% with the genus Rhodocyclus). Based on phenotypic and phylogenetic evidence, it is proposed that strain Ben 117T be designated a novel species in a new genus, Quadricoccus australiensis gen. nov., sp. nov.; the type strain is Ben 117T (= NCIMB 13738T = CIP 107055T). PMID- 11858149 TI - Underwater measurements and modeling of a sonic boom. AB - During a sea trial on the Scotian Shelf, acoustic signals from a sonic boom were recorded on 11 hydrophones of a vertical array. The array spanned the lower 50 m of the water column above a sand bank at 76 m water depth. The source of the sonic boom was deduced to be a Concorde supersonic airliner traveling at about Mach 2. The waterborne waveform was observed to decay as an evanescent wave below the sea surface, as expected. The calm weather (sea state 1) resulted in low ambient noise and low self-noise at the hydrophones, and good signal-to-noise ratio on the upper hydrophones; however, the decreased signal amplitude is more difficult to detect towards the lower part of the water column. The period of the observed waveform is of the order 0.23 s, corresponding to a peak frequency of about 3 Hz. The shape of the measured waveform differs noticeably from the theoretical N-shape waveform predicted with Sawyers' theory [J. Acoust. Soc. Am. 44, 523-524 (1968)]. A simple shallow-ocean geoacoustic model suggests that this effect may be caused in part by seismo-acoustic interaction of the infrasonic waves with the elastic sediments that form the seabed. PMID- 11858150 TI - Anthelmintic resistance in sheep worms in New South Wales, Australia. PMID- 11858151 TI - Malfunction of endocardial defibrillator leads and lead extraction: where do they meet? AB - AIMS: The development of new extraction techniques has improved the success rate of intravascular extraction of pacemaker and defibrillator leads, and hence the practice of extraction is expanding. However, the indications for lead extraction of malfunctioning leads in patients with an implantable cardioverter defibrillator (ICD) are still not well established. METHODS AND RESULTS: We reviewed the literature concerning structural complications of ICD leads. The clinical presentation and detection of malfunction is discussed as well as the consequences for adequate defibrillation therapy. An overview of the current published experience of intravascular extraction of ICD leads is provided including a brief discussion of our own experience. CONCLUSION: From this overview we conclude that malfunctioning pace-sense or ICD leads can be left in situ if there are no uncovered insulation defects. Inserting a new pace-sense or ICD lead is preferable in this situation given the current known complication rate of lead extraction. Lead extraction should be reserved for damaged leads in which interference with proper detection or defibrillation of newly inserted leads cannot be excluded. PMID- 11858152 TI - Task Force on Sudden Cardiac Death, European Society of Cardiology. AB - The European Society of Cardiology has convened a Task Force on Sudden Cardiac Death in order to provide a comprehensive, educational document on this important topic. The main document has been published in the European Heart Journal in August 2001. The Task Force has now summarized the most important clinical issues on sudden cardiac death and provided tables with recommendations for risk stratification and for prophylaxis of sudden cardiac death. The present recommendations are specifically intended to encourage the development and revision of national guidelines on prevention of sudden cardiac death. The common challenge for cardiologists, physicians of other medical specialties and health professionals throughout Europe is to realize the potential for sudden cardiac death prevention and to contribute to public health efforts to reduce its burden. PMID- 11858153 TI - What is the optimal electrode configuration for atrial defibrillators in man? AB - AIMS: To compare the atrial defibrillation threshold (DFT) for two electrode configurations in patients with drug refractory persistent atrial fibrillation (AF). METHODS AND RESULTS: 11 patients, 73% male, mean age 60.9 (range 38 to 83), underwent implantation of a Medtronic Jewel AF dual chamber defibrillator (model 7250). A step-up atrial DFT was performed in a randomized sequence for two electrode configurations: (1) Right atrial to distal coronary sinus electrode (RA > CS) and (2) defibrillator can to right ventricular and right atrial electrodes (CAN > RV + RA). The RA > CS configuration restored SR in 10 patients (91%). The CAN > RA + RV configuration restored SR in four patients (36%). The mean atrial DFT was significantly lower for the RA > CS than CAN > RA + RV configuration (10 +/- 7 Joules vs 25 +/- 6 Joules), P < 0.01. At 3 months post implantation, AF was reinduced and the protocol was repeated for the optimal electrode configuration. There was no significant difference in the atrial DFT compared with that at implant. CONCLUSION: The right atrium to coronary sinus electrode configuration significantly reduces the atrial DFT. The atrial DFT also remains stable at 3 months post-implantation. Patients with persistent AF undergoing insertion of an atrial defibrillator should have a coronary sinus electrode implanted. PMID- 11858154 TI - Deterioration of left ventricular function following atrio-ventricular node ablation and right ventricular apical pacing in patients with permanent atrial fibrillation. AB - AIMS: Transcatheter radiofrequency ablation of the atrio-ventricular (AV) node followed by ventricular pacing has been shown to improve symptoms and quality of life of patients with atrial fibrillation (AF). It is assumed that function improves, but this has been less well demonstrated. The aim of this study was to assess the long-term effect of AV node ablation and ventricular pacing on left ventricular ejection fraction (LVEF) in patients with permanent AF. METHODS AND RESULTS: All 12 patients studied had permanent AF for at least 12 months (mean age 70 years, range 41 to 78). LVEF was determined 6 days and 3 months after AV node ablation by radionuclide ventriculography, at a paced rate of 80 beats . min (-1). Cardiac dimensions were measured by means of transthoracic echocardiography. No major changes in pharmacological therapy were made during 3 months follow-up period. LVEF showed a significant deterioration after 3 months follow-up period for the group (47.5 +/- 14.4%; 6 days after ablation vs 43.2 +/- 13.7%; 3 months after ablation, P < 0.05). There were no significant differences in left ventricular cavity dimensions directly after AV node ablation and 3 months later (LVEDD 51.2 +/- 10.7 mm vs 52.6 +/- 8.6 mm, P = NS: LVESD: 36.1 +/- 14.2 mm vs 36.6 +/- 9.7 mm, P = NS). Left atrial size did not show reduction 3 months after AV node ablation (50.8 +/- 13.6 mm vs 51.0 +/- 14.1 mm, P = NS). CONCLUSION: The restoration of a regular ventricular rhythm following AV node ablation for patients in permanent AF does not result in improvement in left ventricular function. PMID- 11858155 TI - Use of a laser sheath to obtain venous access in pacemaker lead-related obstruction without extraction of the lead. AB - AIMS: Occlusion of the subclavian vein resulting from pacemaker leads prohibits insertion of new leads. We describe the ipsilateral insertion of a new lead without extracting the old lead using a laser sheath in a pacemaker patient with an obstructed vein. METHODS AND RESULT: A laser sheath together with an outer sheath were advanced over the malfunctioning lead just beyond the occlusion. The laser sheath was pulled back and a guide wire inserted through the outer sheath kept in position distal of the occlusion. After removal of the outer sheath a peel-away sheath was introduced and a new lead implanted next to the malfunctioning lead that was abandoned and not extracted. CONCLUSION: By avoiding using the laser along the whole length of the lead we greatly reduced the risk of the procedure but were still able to recanalize the obstructed vein. A risk of bilateral occlusion is avoided and the contralateral site saved as an entry point for future needs. PMID- 11858156 TI - [Echinococcoses]. AB - Human echinococcoses, cystic echinococcosis and alveolar echinococcosis are due to infections with the cestodes Echinococcus granulosus and E. multilocularis, respectively. Both zoonoses share a prolonged latency period before clinical presentation. However their evolution is fairly different: that of a begin tumor of the liver or lung for cystic echinoccocosis, and that of a slowly developing malignant tumor of the liver for alveolar echinoccocosis, with subsequent invasion of liver vessels and bile ducts and metastatic dissemination. Ultrasonography, CT-scan and specific serology are the key-exams for diagnosis. In both forms, surgery is the treatment of choice when a complete resection is possible. Liver transplantation may be an ultimate treatment option in very advanced cases of alveolar echinoccocosis. However, alternative treatment procedures have been proposed in the past 15 years and, combined with an earlier diagnosis, they have markedly improved patients survival and quality of life. Interventional radiology (puncture, aspiration, injection, reaspiration) has become a fully validated treatment of cystic echinoccocosis, and may be used in alveolar echinoccocosis for alleviating some of the complications of the disease such as biliary obstruction or bacterial superinfection. Albendazole, at high dosage, is a necessary complementary treatment after any intervention procedure, and for life when radical resection is not possible. Prevention relies on personal measures of hygiene and heating of contaminated food, and on collective measures aimed at reducing cestode egg shedding by the feces of infected canivores. PMID- 11858157 TI - Genomic sequence and cardiac expression of atrial natriuretic peptide in cats. AB - OBJECTIVE: To determine the nucleotide and amino acid sequence of atrial natriuretic peptide (ANP) in cats and its typical regions of cardiac expression. ANIMALS: 5 healthy adult mixed-breed cats. PROCEDURE: Total RNA was extracted from samples obtained from the left and right atrium, left and right ventricle, and interventricular septum of each cat. The RNA was used to produce cDNA for sequencing and northern blot analysis. Genomic DNA was extracted from feline blood samples. Polymerase chain reaction primers designed from consensus sequences of other species were used to clone and sequence the feline ANP gene. RESULTS: The feline ANP gene consists of 1,072 nucleotides. It consists of 3 exons (123, 327, and 12 nucleotides) separated by 2 introns (101 and 509 nucleotides). It has several typical features of eukaryotic genes and a putative steroid-response element located within the second intron. Preprohormone ANP consists of 153 amino acids. The amino acid sequence of the active form of feline ANP (ANP-30) is identical to that of equine, bovine, and ovine ANP-30 and differs from that of human, canine, and porcine ANP-28 only by 2 carboxy-terminal arginine residues. The ANP mRNA was detected only in the left and right atria. CONCLUSIONS AND CLINICAL RELEVANCE: The genetic and protein structure and principal regions of cardiac expression of feline ANP are similar to those of other species. Results of this study should be helpful in future studies on the natriuretic response in cats to diseases that affect cardiovascular function. PMID- 11858158 TI - Do interleukin-1 polymorphisms predict the development of periodontitis or the success of dental implants? AB - Factors which increase the risk of severe adult periodontitis (AP) may also contribute to the success of dental implants. To determine which cytokines may be relevant, levels of interleukin-1alpha (IL-1alpha), interleukin-1beta (IL-1beta), interleukin-1 receptor antagonist (IL-1ra), interleukin-6 (IL-6) and interferon gamma (IFN-gamma) mRNA were quantitated in gingival tissue from periodontitis patients and healthy controls. Periodontitis significantly increased levels of IL 1alpha, IL-1beta, IL-6 and IFN-gamma mRNA relative to healthy tissues. IL-1 was selected for further study, as it has inflammatory and bone resorbing properties. We examined IL-1A(-889) and IL-1B(+3953) alleles in Caucasian patients with AP and early-onset periodontitis (EOP), patients with dental implants and healthy individuals. The IL-1B(+3953) polymorphism was associated with AP. This was evident from an increased homozygosity for allele 2 in patients with AP and a decreased heterozygosity in advanced AP patients. IL-1A(-889) and a composite genotype [IL-1A(-889)2 plus IL-1B(+3953)2] showed no association with the incidence of periodontitis, disease onset or disease severity. IL-1A(-889), IL 1B(+3953) and the composite genotype also showed no association with failure of dental implants. PMID- 11858159 TI - Validity of the Index of Complexity, Outcome, and Need (ICON) in determining orthodontic treatment need. AB - Occlusal indices are used to determine eligibility for orthodontic treatment in several publicly funded programs. The Index of Complexity, Outcome, and Need (ICON), based on the perception of 97 orthodontists from 9 countries, has been proposed as a multipurpose occlusal index. The aim of this study was to investigate the validity of the ICON as an index of orthodontic treatment need compared with the perception of need as determined by a panel of US orthodontists. One hundred seventy study casts, representing a full spectrum of malocclusion types and severity, were scored for orthodontic treatment need by an examiner calibrated in the ICON. The results were compared with the decisions of an expert panel of 15 orthodontic specialists from the central Ohio area. The simple kappa statistic (0.81) indicated very high agreement of the index with the decisions of the expert panel. The sensitivity (94%), specificity (85%), positive predictive value (92%), negative predictive value (90%), and overall accuracy of the ICON (91%) also confirmed good agreement with the orthodontic specialists. The panel found that 64% of the casts required orthodontic treatment; the ICON scores indicated that 65% of the cases needed treatment. There was agreement between the expert panel and the index in 155 of the 170 cases. These results support the use of the ICON as a validated index of orthodontic treatment need. PMID- 11858160 TI - AIDS herbal therapy. PMID- 11858161 TI - Historical highlights in bionics and related medicine. PMID- 11858162 TI - Editorial: Septolateral cinching. PMID- 11858163 TI - Outcome after mitral valve repair for functional ischemic mitral regurgitation. AB - BACKGROUND AND AIM OF THE STUDY: Functional ischemic mitral regurgitation (MR) can occur secondary to coronary artery disease. Controversy exists regarding management of these patients. Mitral valve annuloplasty in conjunction with coronary artery bypass grafting (CABG), accepted as the best treatment for severe MR, has been disputed for lesser degrees of regurgitation due to higher mortality. The results of a combined procedure approach were reviewed. METHODS: Between February 1992 and June 1999, 100 consecutive patients (mean age 67+/-11 years) with functional ischemic MR underwent mitral valve repair + CABG. The repair was limited to a Duran flexible annuloplasty ring. Among patients, 72% had a preoperative myocardial infarction and 51% required perioperative intra-aortic balloon pump. NYHA functional class was III-IV in 72%; preoperative MR by transesophageal echocardiography (TEE) was grade 3-4+ in 80% and grade 2+ in 20%. RESULTS: Intraoperative completion TEE indicated 0-1+ MR in 98%. Early mortality was 12% and late mortality 14%, for an overall survival of 74%. The mean follow up was 35.8 months. Follow up TEE on 82% of patients showed zero to trivial MR in 42% of patients, grade 1+ MR in 29%, 2+ MR in 24%, and 3-4+ MR in 5%. Follow up NYHA class was I-II in 81% of patients and III-IV in 19%. A significant correlation was found between recurrent MR and declining left ventricular function on follow up only, as well as the occurrence of preoperative myocardial infarction. CONCLUSION: Functional ischemic MR remains a difficult problem to treat, and has a poor long-term outcome. Ring annuloplasty for functional ischemic MR with coronary artery disease achieves immediate valve competence. However, a significant number of patients develop recurrent MR at intermediate follow up. PMID- 11858164 TI - Torsion dynamics in the evolution from acute to chronic mitral regurgitation. AB - BACKGROUND AND AIM OF STUDY: Left ventricular (LV) torsion reduces transmural fiber strain gradients during systole, and torsional recoil in early diastole is thought to assist LV filling. To test the hypothesis that deterioration of torsional dynamics accompanied LV dysfunction during the evolution of mitral regurgitation (MR), torsion was measured during the progression from acute to chronic MR in a canine model. METHODS: Seven dogs underwent cardiopulmonary bypass for LV marker placement and creation of MR by disrupting the posterior leaflet. After 7-10 days, three-dimensional marker coordinates were measured with biplane videofluoroscopy to study LV geometry, size and function, plus maximal torsional deformation, time of maximal torsion relative to end-ejection, and early diastolic torsional recoil during the first 5% of filling. After three months, the animals were re-studied. RESULTS: Progression from acute to chronic MR was associated with a significant decrease in maximum LV dP/dt (1,574+/-213 to 1,300+/-252 mmHg/s, p <0.01) and an increase in LVEDP from 11+/-5 to 15+/-5 mmHg (p <0.01). After three months of MR, maximum torsional deformation decreased from 6.3+/-1.9 to 4.7+/-2.0 degrees (p = 0.04), as did early diastolic recoil (-3.8+/ 1.0 to -1.5+/-1.7 degrees, p = 0.03). CONCLUSION: Progression from acute to chronic MR is accompanied by decreased and delayed systolic LV torsional deformation and a decline in early diastolic recoil, which may contribute to LV dysfunction by increasing transmural strain gradients during systole and impairing diastolic filling. As torsional deformation and recoil can be measured non-invasively with MRI in humans, such measurements might prove useful in patients with progressive MR as an adjunct to determine the timing of surgical repair. PMID- 11858165 TI - Dynamics of metal adaptation in riverine chironomids. AB - The ability of the non-biting midge Chironomus riparius to survive and reproduce in metal polluted lowland rivers facilitates the opportunity to study micro evolutionary processes in situ. However, due to larval drift, adapted midge populations are subject to regular immigration of non-adapted specimens from clean upstream river reaches. To examine the influence of non-adapted genes in adapted midge populations on the level of metal adaptation, an upstream and downstream chironomid population were crossbred on eight separate occasions in the laboratory to mimic gene flow. Several life-history characteristics, indicating adaptation to metals, were followed seasonally in the parental strains as well as in the reciprocal crossings. Such crossings were done over a 14-month period and maternal effects were found to be absent, indicating a major genetic component for the increased metal tolerance in the exposed midge populations. Furthermore, results confirmed the presence of adaptation to metals in exposed chironomids. However, a rapid loss of metal adaptation in the first generation hybrid offspring was clearly demonstrated. Consequently, the large temporal variation in metal adaptation in midge populations from the river can be explained by the earlier reported seasonal variations in selection pressure and immigration rates from non-adapted sub-populations. PMID- 11858166 TI - Matrix effects in applying mono- and polyclonal ELISA systems to the analysis of weathered oils in contaminated soil. AB - Commercial mono- and polyclonal enzyme-linked immunosorbent assay (ELISA) systems were applied to the on-site analysis of weathered hydrocarbon-contaminated soils at a former integrated steelworks. Comparisons were made between concentrations of solvent extractable matter (SEM) determined gravimetrically by Soxhlet (dichloromethane) extraction and those estimated immunologically by ELISA determination over a concentration range of 2000-330,000 mg SEM/kg soil dry weight. Both ELISA systems tinder-reported for the more weathered soil samples. Results suggest this is due to matrix effects in the sample rather than any inherent bias in the ELISA systems and it is concluded that, for weathered hydrocarbons typical of steelworks and coke production sites, the use of ELISA requires careful consideration as a field technique. Consideration of the target analyte relative to the composition of the hydrocarbon waste encountered appears critical. PMID- 11858167 TI - Bachmann's bundle pacing: a role for three-dimensional intracardiac echocardiography? PMID- 11858168 TI - Incidence of acute pancreatitis. PMID- 11858169 TI - Expression and immunolocalization of heat shock proteins in the healing of gastric ulcers in rats. AB - BACKGROUND: Heat shock proteins are induced when cells are subjected to noxious stimuli. They afford cytoprotection and increase the resistance of the tissue to damage. However, their roles in the healing of gastric ulcers have not been well established. In this study, the expression and immunolocalization of three heat shock proteins (HSPs); namely inducible HSP70 (iHSP70), HSP47, and HSP32 during ulcer healing were investigated in rats with gastric ulcer. METHODS: Gastric ulcers (kissing ulcers) were induced by luminal application of acetic acid solution. Gastric tissue samples were obtained from the ulcer base, ulcer margin, and non-ulcerated area around the ulcer margin at different time intervals after ulcer induction. The protein levels and distributions of HSPs were analyzed with Western blotting and immunohistochemical methods, respectively. RESULTS: It was found that all HSPs were expressed in normal, non-ulcerated, and gastric ulcer tissues. HSP32 was elevated during inflammation (1-8 days after ulcer induction), while HSP47 expression was exacerbated at the ulcer base and margin during ulcer healing (3-12 days). Decreased expression of iHSP70 was observed at the ulcer base immediately after ulcer induction, but returned to normal level by the end of the healing stage (8-12 days). Inducible HSP70 was found distributed in the gastric glands and injured tissues in the inflamed areas. Wide distribution of HSP47 was detected in granulation tissues and collagen producing cells, while HSP32 was localized in the gastric glands and inflammatory cells. CONCLUSIONS: The findings indicate that iHSP70, HSP47, and HSP32 play different roles during ulcer healing. HSP32 seems to act as an inflammatory defensive factor, and HSP47 as a collagen-specific molecular chaperon contributing significantly to gastric ulcer healing. However, the role of iHSP70 in the ulcer healing process is still undefined. PMID- 11858170 TI - N-acetyltransferase 2 polymorphism is not related to the risk of advanced alcoholic liver disease. AB - BACKGROUND: Ethanol abuse is the most prevalent cause of liver cirrhosis in Spain. Genetic polymorphisms affect the activity of the enzymes involved in ethanol metabolism and in processing the toxic by-products generated in the liver. N-acetyltransferase 2 (NAT2) is a polymorphic phase 2 enzyme not involved in these processes, but recent data suggest that the most prevalent slow acetylator genotype protects against the risk of advanced alcoholic liver disease (ALD). We have identified six single nucleotide polymorphisms (SNP) at the NAT2 gene locus in order to disclose whether such an association exists. METHODS: Genomic DNA from 95 ALD patients (15 with superimposed hepatocellular carcinoma (HCC)) and from 258 healthy individuals was analysed for SNPs at the coding region of the NAT2 gene by means of allele-specific polymerase chain reaction. RESULTS: There are no differences in the relative frequencies of the eight identified NAT2 alleles (including the wild-type allele) nor in the distribution of predicted phenotypes (54% of slow acetylators in each group). Twelve patients with HCC (80%) were slow acetylators (P < 0.05). CONCLUSIONS: There is no relationship between the NAT2 genotype and the risk of ALD. Slow acetylator genotype may predispose to the development of HCC in severe ALD patients not infected by the hepatitis C virus. PMID- 11858171 TI - Differential effectiveness of salicylate-dependent and jasmonate/ethylene dependent induced resistance in Arabidopsis. AB - Salicylic acid (SA), jasmonic acid (JA), and ethylene (ET) are each involved in the regulation of basal resistance against different pathogens. These three signals play important roles in induced resistance as well. SA is a key regulator of pathogen-induced systemic acquired resistance (SAR), whereas JA and ET are required for rhizobacteria-mediated induced systemic resistance (ISR). Both types of induced resistance are effective against a broad spectrum of pathogens. In this study, we compared the spectrum of effectiveness of SAR and ISR using an oomycete, a fungal, a bacterial, and a viral pathogen. In noninduced Arabidopsis plants, these pathogens are primarily resisted through either SA-dependent basal resistance (Peronospora parasitica and Turnip crinkle virus [TCV]), JA/ET dependent basal resistance responses (Alternaria brassicicola), or a combination of SA-, JA-, and ET-dependent defenses (Xanthomonas campestris pv. armoraciae). Activation of ISR resulted in a significant level of protection against A. brassicicola, whereas SAR was ineffective against this pathogen. Conversely, activation of SAR resulted in a high level of protection against P. parasitica and TCV, whereas ISR conferred only weak and no protection against P. parasitica and TCV, respectively. Induction of SAR and ISR was equally effective against X. campestris pv. armoraciae. These results indicate that SAR is effective against pathogens that in noninduced plants are resisted through SA-dependent defenses, whereas ISR is effective against pathogens that in noninduced plants are resisted through JA/ET-dependent defenses. This suggests that SAR and ISR constitute a reinforcement of extant SA- or JA/ET-dependent basal defense responses, respectively. PMID- 11858172 TI - Overexpression of the tomato Asc-1 gene mediates high insensitivity to AAL toxins and fumonisin B1 in tomato hairy roots and confers resistance to Alternaria alternata f. sp. lycopersici in Nicotiana umbratica plants. AB - The sphinganine-analog mycotoxins (SAMs) fumonisin B1 and AAL toxins are inhibitors of eukaryotic sphinganine N-acyltransferase in vitro. Treatment of eukaryotes with SAMs generally results in an accumulation of sphingoid base precursors and a depletion of complex sphingolipids. The asc,asc genotypes of tomato (Lycopersicon esculentum) and Nicotiana umbratica are sensitive to SAMs and host of the AAL toxin-producing fungus Alternaria alternata f. sp. lycopersici. Codominant insensitivity to SAMs in tomato is mediated by the Asc-1 gene, and sensitivity is associated with a frame-shift mutation present in asc-1. We investigated the function of Asc-1 in mediating insensitivity to SAMs and resistance to the fungus by overexpression of asc-1 and Asc-1. In this study, it is shown that overexpression of these genes did not lead to visual symptoms in tomato hairy roots and N. umbratica plants. Overexpression of asc-1 did not influence the (in)sensitivity to SAMs. Overexpression of Asc-1 in SAM-sensitive hairy roots and N. umbratica plants, however, mediated a high insensitivity to SAMs and resistance to plant infection by Alternaria alternata f. sp. lycopersici. PMID- 11858173 TI - dpp genes of Rhizobium leguminosarum specify uptake of delta-aminolevulinic acid. AB - An operon with homology to the dppABCDF genes required to transport dipeptides in bacteria was identified in the N2-fixing symbiont, Rhizobium leguminosarum. As in other bacteria, dpp mutants were severely affected in the import of delta aminolevulinic acid (ALA), a heme precursor. ALA uptake was antagonized by adding dipeptides, indicating that these two classes of molecule share the same transporter. Mutations in dppABCDF did not affect symbiotic N2 fixation on peas, suggesting that the ALA needed for heme synthesis is not supplied by the plant or that another uptake system functions in the bacteroids. The dppABCDF operon of R. leguminosarum resembles that in other bacteria, with a gap between dppA and dppB containing inverted repeats that may stabilize mRNA and may explain why transcription of dppA alone was higher than that of dppBCDF. The dppABCDF promoter was mapped and is most likely recognized by sigma70. PMID- 11858174 TI - Chemically induced virus resistance in Arabidopsis thaliana is independent of pathogenesis-related protein expression and the NPR1 gene. AB - Salicylic acid (SA) treatment triggers inhibition of replication or movement of several positive-sense RNA plant viruses in tobacco. This resistance can also be stimulated by nonlethal concentrations of cyanide and antimycin A (AA) without triggering induction of pathogenesis-related PR-1 protein genes. In two ecotypes of Arabidopsis thaliana (Columbia and Nossen), SA-induced resistance to a tobamovirus, Turnip vein clearing virus (TVCV), was also induced by nonlethal concentrations of cyanide and AA without concomitant induction of PR-1 gene expression. Furthermore, chemically induced resistance to TVCV, as well as the induction of the plant mitochondrial alternative oxidase (a potential target for the chemicals), was independent of NPR1, a gene that plays a key role downstream of SA in the induction of PR proteins. The chemically induced resistance to TVCV appeared to be due to inhibition of replication at the site of inoculation. Taken together, these results show that in Arabidopsis, as in tobacco, resistance to viruses can be induced via a distinct branch of the defensive signal transduction pathway. This suggests that the existence of this virus-specific branch may be widespread among plants. PMID- 11858175 TI - Self-assembly of tetrahedral and trigonal antiprismatic clusters [Fe4(L4)4] and [Fe6(L5)6] on the basis of trigonal tris-bidentate chelators. AB - In a one-pot reaction, the tetranuclear iron chelate complex [Fe4(L4)4] 6 was generated from benzene-1,3,5-tricarboxylic acid trichloride (4), bis-tert-butyl malonate (5a), methyllithium, and iron(II) dichloride under aerobic conditions. Alternatively, hexanuclear iron chelate complex [Fe(L5)6] 7 was formed starting from bis-para-tolyl malonate (5b) by employing identical reaction conditions to those applied for the synthesis of 6. The clusters 6 and 7 are present as racemic mixtures of homoconfigurational (delta,delta,delta,delta)/(lambda,lambda,lambda,lambda)-fac or (delta,delta,delta,delta,delta,delta)/(lambda,lambda,lambda,lambda,lambda,lambda) fac stereoisomers. The structures of 6 and 7 were unequivocally resolved by single-crystal X-ray analyses. The all-iron(III) character of 6 and 7 was determined by Mossbauer spectroscopy. PMID- 11858176 TI - Maternal pulmonary adenocarcinoma metastatic to the fetus: first recorded case report and literature review. AB - We report the first maternal pulmonary adenocarcinoma metastatic to the fetus as well as an updated literature review. Review of the literature revealed that there have been only 67 cases of maternal malignancy metastatic to the products of conception. These were mostly malignant melanoma and hematopoietic tumors. A 46-year old multiparous woman with metastatic pulmonary adenocarcinoma, diagnosed at 23 weeks gestation, delivered a male infant who appeared normal at birth. The mother died 2 days after delivery. The child developed multiple scalp tumors a 2 weeks of age. The tumors recurred rapidly after initial resection. Wide local excision of the involved scalp and skin graft coverage was performed at 14 weeks of age. Histopathology of these tumors was identical to that of the maternal tumor. The maternal origin of these tumors was confirmed by fluorescence in situ hybridization (FISH). The child is now 5 years old and free of disease. PMID- 11858177 TI - The effects of visual scenes on roll and pitch thresholds in pilots versus nonpilots. AB - BACKGROUND: Previous studies have indicated that, compared with nonpilots, pilots rely more on vision than "seat-of-the-pants" sensations when presented with visual-vestibular conflict. The objective of this study was to evaluate whether pilots and nonpilots differ in their thresholds for tilt perception while viewing visual scenes depicting simulated flight. METHODS: This study was conducted in the Advanced Spatial Disorientation Demonstrator (ASDD) at Brooks AFB, TX. There were 14 subjects (7 pilots and 7 nonpilots) who recorded tilt detection thresholds in pitch and roll while exposed to sub-threshold movement in each axis. During each test run, subjects were presented with computer-generated visual scenes depicting accelerating forward flight by day or night, and a blank (control) condition. RESULTS: The only significant effect detected by an analysis of variance (ANOVA) was that all subjects were more sensitive to tilt in roll than in pitch [F (2,24) = 18.96, p < 0.001]. Overall, pilots had marginally higher tilt detection thresholds compared with nonpilots (p = 0.055), but the type of visual scene had no significant effect on thresholds. CONCLUSION: In this study, pilots did not demonstrate greater visual dominance over vestibular and proprioceptive cues than nonpilots, but appeared to have higher pitch and roll thresholds overall. The finding of significantly lower detection thresholds in the roll axis vs. the pitch axis was an incidental finding for both subject groups. PMID- 11858178 TI - Response to bioterrorism. Terror weapons are regarded as weapons of mass destruction. PMID- 11858179 TI - Response to bioterrorism. US anthrax incidents led to scares in Scotland. PMID- 11858180 TI - Response to bioterrorism. Screening for agents of bioterrorism increases terror. PMID- 11858181 TI - Response to bioterrorism. Countermeasures against weapons of mass destruction must be assessed now. PMID- 11858182 TI - Response to bioterrorism. Anthrax issue underlines need for infection specialists trained at bedside. PMID- 11858184 TI - Enhanced PAI-1 levels from early second trimester and during labour and plasminogen activators in normal pregnancy. PMID- 11858183 TI - Mechanism of resveratrol-mediated suppression of tissue factor gene expression. AB - Tissue factor (TF) is a cell surface receptor for factor VII(a), and the binding of factor VII(a) to TF initiates the coagulation cascade. Inappropriate in vivo expression of TF in vascular cells has been shown to be responsible for thrombotic disorders associated with a variety of pathological conditions, including gram-negative sepsis, cancer and atherosclerosis. A number of epidemiological studies suggest that moderate consumption of red wine provides protective effects against coronary heart disease mortality. Recently, we have shown that resveratrol, a polyphenolic compound found in wine, inhibited the induction of TF expression in endothelial cells and mononuclear cells (Pendurthi UR, Williams JT, Rao LVM. Arterioscler Thromb Vasc Biol 1999: 19: 419-426). In the present study, we examined the mechanism by which resveratrol inhibits the expression of TF in monocytes by using a monocytic cell line, THP-1, as a model cell. Northern blot analysis, gel mobility shift assays and transfection studies with various TF promoter constructs, as well as other transcription regulatory constructs, were used to elucidate the inhibitory mechanism of resveratrol. The data show that resveratrol inhibited lipopolysaccharide (LPS)-induced expression of TF in human monocytes and monocytic cell line, THP-1 in a dose dependent manner. Resveratrol did not significantly alter the binding of various transcription factors involved in TF gene expression to DNA. However, resveratrol suppressed the transcription of cloned human TF promoter. Further experiments revealed that resveratrol reduced kappaB- but not AP-1-driven transcriptional activity. Additional experiments showed that resveratrol suppressed the phosphorylation of p65 and its transactivation. In summary, our results indicate that resveratrol does not inhibit the activation or translocation of NF kappaB/Rel proteins but inhibits NF-kappaB/Rel-dependent transcription by impairing the transactivation potential of p65. PMID- 11858185 TI - Neurohormonal activation does not explain elevated tissue factor expression in heart failure. PMID- 11858186 TI - The effect of fibrinogen genotype on fibrinogen levels after strenuous physical exercise. AB - We have examined the effect of two beta-fibrinogen gene promoter polymorphisms ( 455G>A and -854G>A) on the fibrinogen response to severe exercise in a group of male army recruits undergoing basic training. Fibrinogen was measured pre training and again serially after severe 48 h final military exercise (FME). Out of 884 subjects, 762 completed training of whom 250 were selected for post-FME study. Fibrinogen levels (g/l) were significantly elevated over baseline levels 2, 48 and 96 h after FME, representing increases of 15.7%, 3.4% and 7.6% (p <0.005; p = 0.05 and p <0.005 respectively), with higher levels in -455A allele carriers than genotype -455GG: 3.17+/-0.05 vs. 2.94+/-0.05 (p <0.001), 2.86+/ 0.05 vs. 2.60+/-0.05 (p <0.0005) and 2.98+/-0.06 vs. 2.69+/-0.06 (p <0.0005) at 2, 48 and 96 h respectively. There was no effect of the -854G>A polymorphism on fibrinogen, even after taking into account beta-fibrinogen -455 genotype. Thus the fibrinogen -455G>A polymorphism influences fibrinogen levels following exercise. The effect of genotype might be clinically relevant at times of hyperfibrinogenaemia such as following an acute inflammatory response. PMID- 11858187 TI - The role of cytokines in activation of coagulation and fibrinolysis in dengue shock syndrome. AB - In a prospective clinical study of 50 patients with Dengue Shock Syndrome (DSS), we investigated the association of tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta (IL-1beta), IL-1 receptor antagonist (IL-1Ra), and IL-6 with activation markers of coagulation (F1+2 and TATc) and fibrinolysis (t-PA, PAPc, and D-dimer). We found that TNF-alpha, IL-1beta and Il-1Ra, but not IL-6, concentrations were elevated in the circulation during the early stage of infection and at discharge from hospital. TNF-alpha was significantly associated with D-dimer, an activation marker of fibrinolysis (p < 0.003), but not with activation markers of coagulation. IL-1beta was significantly associated with t PA (p < 0.03). IL-1Ra was significantly associated with F1+2, TATc (p < 0.04 and p < 0.02, respectively), whereas IL-6 was significantly associated with both, activation markers of coagulation (F1+2; p < 0.03) and fibrinolysis (PAPc; p = 0.002). Our data are in line with studies in bacterial sepsis. In severe dengue virus infection the same cytokines are involved in the onset and regulation of hemostasis. PMID- 11858188 TI - D-dimer as a risk factor for deep vein thrombosis: the Leiden Thrombophilia Study. AB - We studied the association of D-dimer with the risk of deep vein thrombosis (DVT). D-dimer was measured in 474 patients more than 6 months after diagnosis of a first DVT and in 474 age- and sex-matched controls. For D-dimer above the 70th percentile (130.5 ng/ml), the odds ratio (OR) for DVT was 2.2 (95% CI, 1.6-2.9). The association was unchanged with adjustment for other risk factors. Excluding participants with Factor V Leiden, prothrombin 20210A, or factors VIIIc or IX above the 90th percentile, the OR was 1.6 (95% CI, 1.1-2.3). The risks of DVT with the joint presence of high D-dimer and either factor V Leiden or prothrombin 20210A were increased 12.4-fold (95% CI 5.6-27.7) and 7.2-fold (95% CI 2.1-25.1), respectively. Higher D-dimer concentration was associated with the risk of DVT, and was supra-additive to the risks associated with factor V Leiden and the prothrombin 20210A variant. Persistence of this association in the absence of other hemostatic risk factors for DVT suggests that high D-dimer may be related to other, as yet unknown, risk factors for venous thrombosis. Confirmation of these findings is desirable. PMID- 11858189 TI - Early antithymocyte globulin therapy improves survival in patients with steroid resistant acute graft-versus-host disease. AB - Second-line therapies for steroid-resistant acute GVHD have been used with limited success. We have reviewed the responses of 79 hematopoetic stem cell transplant (HSCT) patients uniformly treated from 1990-1998 with equine antithymocyte globulin (ATG) for steroid-resistant acute GVHD, defined as progression of acute GVHD after 4 days of treatment with prednisone or no improvement of acute GVHD after 7 days of treatment with prednisone. Patients received HSCT from 34 related (32 matched sibling/2 partially matched) and 45 unrelated (14 HLA-A, -B, -DRB1 matched/31 partially matched) donors. Prior to ATG therapy, severe (grade III-IV) GVHD was observed in 34 patients (43%). Organs involved included skin in 81% of patients, lower GI tract in 52%, upper GI tract in 28%, and liver in 11%. Treatment consisted of 1-5 courses (median, 2 courses) of ATG (15 mg/kg per dose bid x 5 days) given for a median of 16 days (range, 5 to 44 days) after the onset of GVHD. All patients continued to receive prednisone, 60 mg/m2 per day (or methylprednisolone IV equivalent), plus CSA (75%) or tacrolimus (4%). At day 28 of treatment, overall improvement was observed in 54% of patients; durable (> or = 28 days) complete response was observed in 20% of patients, and partial response was observed in 34% of patients. In multivariate analysis, patients with CML or a malignant disease other than acute leukemia had a greater likelihood of overall response than did those with nonmalignant diseases. Patients with acute skin GVHD (with or without other organ involvement) responded most frequently. Chronic GVHD developed in 51% of patients by 1 year after HSCT. One patient developed EBV lymphoproliferative disease. For the entire cohort, the probability of survival at 1 year was 32% (95% CI, 22%-42%). In multivariate analysis, factors associated with better survival included earlier onset of acute GVHD, shorter time from initial treatment for GVHD to treatment with ATG, and the use of non-T-cell-depleted stem cell grafts. These data suggest that treatment with ATG can be an active therapy, especially in patients with skin GVHD and early signs of steroid resistance. PMID- 11858190 TI - Morbidity and mortality of chronic GVHD after hematopoietic stem cell transplantation from HLA-identical siblings for patients with aplastic or refractory anemias. AB - We analyzed effects of successive changes in prevention and treatment of chronic GVHD in 405 patients with aplastic anemia and refractory anemia given HLA-matched hematopoietic stem cell transplantation (HSCT) from 1970-1997. For analysis, patients were divided into group I, transplantations from 1970-1976; group II, 1977-1983; group III, 1984-1990; and group IV, 1991-1997. The overall incidence of chronic GVHD was 28%. Incidences of chronic GVHD for groups I through IV were 20%, 46%, 41%, and 22%, respectively, reflecting added buffy coat infusions in groups II and III. Five-year survival rates of patients with chronic GVHD for groups I through IV were 58%, 74%, 82%, and 76%, respectively (NS). Among group I patients, 50% were alive off immunosuppression, none were alive on immunosuppression, and 50% died. These figures were 76%, 0%, and 24% in group II; 80%, 10%, and 10% in group III; and 64%, 21%, and 14% in group IV patients. More serious infections and skin contractures were seen in group I than in groups II, III, and IV (P = .0001, .02, .01 and P =.0003, .001, .05, respectively). Lung complications, aseptic necroses, depression, and Karnofsky scores were comparable among groups. Gastrointestinal complications seemed less frequent among groups II through IV. Diabetes mellitus was more frequent in group IV than in groups I through III (P = .008). Secondary malignancies occurred in 33%, 6%, 3%, and 0% of patients in the 4 groups, respectively. In conclusion, over 28 years, chronic GVHD has remained challenging, with only slight improvements in quality of life. Decisive improvements in therapy and survival will have to await both a better understanding of the immunological events underlying chronic GVHD and better infection prevention and control. PMID- 11858191 TI - Lifestyle factors as risk factors for fatigue and psychological distress in the working population: prospective results from the Maastricht Cohort Study. AB - We examined potentially modifiable lifestyle factors as possible risk factors for the onset of fatigue and psychological distress after 1-year follow-up among 8833 employees who participated in the prospective Maastricht Cohort Study of "Fatigue at Work." Results showed, even after adjustment for demographics, presence of disease, other lifestyle factors, psychosocial work characteristics, and psychological distress, that overweight (body mass index, 25 to 29.9) and being physically inactive during leisure time were strongly related to onset of fatigue in men, whereas underweight (body mass index, < 18.5) in women increased the risk for future fatigue. In addition, the study suggests some differential effects of lifestyle factors in the onset of psychological distress. Certainly, these modifiable factors can be targeted in interventions, either on an individual or group level, to prevent or at least reduce the risk of developing fatigue and psychological distress in the working population. PMID- 11858192 TI - Gallstones and ethnicity in the Americas. AB - Information on ethnicity as related to gallstones has been limited by insufficient or inaccurate characterization of ethnicity. Nevertheless, in recent years, ultrasonography has allowed limited examination of ethnic differences in the risk of gallbladder disease, defined by a history of cholecystectomy or ultrasonographic detection of gallstones. Among women, the risk of gallbladder disease is highest among American Indians, followed by Hispanics, non-Hispanic whites, and non-Hispanic blacks. Men differ from women by having lower risk in all ethnic groups and by having a similar prevalence between Hispanics and non Hispanic whites. It does not appear that the type of stone differs much according to ethnic group in the United States. Well-known risks for gallbladder disease, such as obesity, weight loss, pregnancy, and low alcohol use do not explain differences in ethnic risk. As yet, genetic markers have not been identified that would explain differences in risk among ethnic groups. Higher case fatality rates among non-Hispanic blacks than non-Hispanic whites suggest that blacks may have inadequate access to medical care for gallbladder disease. PMID- 11858193 TI - Viability of a five-strain mixture of Listeria monocytogenes in vacuum-sealed packages of frankfurters, commercially prepared with and without 2.0 or 3.0% added potassium lactate, during extended storage at 4 and 100 degrees C. AB - The viability of Listeria monocytogenes was monitored on frankfurters containing added potassium lactate that were obtained directly from a commercial manufacturer. Eight links (ca. 56 g each) were transferred aseptically from the original vacuum-sealed bulk packages into nylon-polyethylene bags. Each bag then received a 4-ml portion of a five-strain mixture of the pathogen. Frankfurters containing 2.0 or 3.0% potassium lactate were evaluated using 20 CFU per package, and frankfurters containing 3.0% potassium lactate were evaluated using 500 CFU per package. The packages were vacuum-sealed and stored at 4 or 10 degrees C for up to 90 or 60 days, respectively. During storage at 4 degrees C, pathogen numbers remained at about 1.6 log10 CFU per package over 90 days in packages containing frankfurters with 2.0% potassium lactate that were inoculated with about 20 CFU. In packages containing frankfurters with 3.0% potassium lactate that were inoculated with about 20 CFU and stored at 4 degrees C, pathogen numbers remained at about 1.4 log10 CFU per package over 90 days. In packages containing frankfurters with 3.0% potassium lactate that were inoculated with about 500 CFU and stored at 4 degrees C, pathogen numbers remained at about 2.4 log10 CFU per package over 90 days. However, in the absence of any added potassium lactate, pathogen numbers increased to 4.6 and 5.0 log10 CFU per package after 90 days of storage at 4 degrees C for starting levels of 20 and 500 CFU per package, respectively. During storage at 10 degrees C, pathogen numbers remained at about 1.4 log10 CFU per package over 60 days in packages containing frankfurters with 2.0% potassium lactate that were inoculated with about 20 CFU. In packages containing frankfurters with 3.0% potassium lactate that were inoculated with about 20 CFU and stored at 10 degrees C, pathogen numbers remained at about 1.1 log10 CFU per package over 60 days of storage. In the absence of any added potassium lactate, pathogen numbers increased to 6.5 log10 CFU per package after 28 days and then declined to 5.0 log10 CFU per package after 60 days of storage at 10 degrees C. In packages containing frankfurters with 3.0% potassium lactate that were inoculated with about 500 CFU per package, pathogen numbers remained at about 2.4 log10 CFU per package over 60 days of storage at 10 degrees C, whereas in the absence of any added potassium lactate, pathogen numbers increased to about 6.6 log10 CFU per package within 40 days and then declined to about 5.5 log10 CFU per package after 60 days of storage. The viability of L. monocytogenes in frankfurter packages stored at 4 and 10 degrees C was influenced by the pH and the presence or levels of lactate but not by the presence or levels of indigenous lactic acid bacteria or by the proximate composition of the product. These data establish that the addition of 2.0% (P < 0.0004) or 3.0% (P < 0.0001) potassium lactate as an ingredient in frankfurters can appreciably enhance safety by inhibiting or delaying the growth of L. monocytogenes during storage at refrigeration and abuse temperatures. PMID- 11858194 TI - A polymerase chain reaction-based method for the detection of hepatitis A virus in produce and shellfish. AB - Outbreaks of gastroenteritis that are suspected to be of viral origin are on the rise. Thus, there is a need for regulatory agencies entrusted with food safety to develop adequate techniques for the detection of viruses in foods. We have established a general procedure for the detection of hepatitis A virus (HAV) in shellfish that, with minor modifications, is also applicable to fresh produce such as cilantro. Total RNA was isolated from shellfish or cilantro, followed by isolation of poly(A)-containing RNA. Because HAV genomic RNA contains a poly(A) tail, the isolation of poly(A)-containing RNA also enriches HAV genomic RNA. Reverse transcription was used to convert the RNA to cDNA, and then amplification was carried out by polymerase chain reaction (PCR). Reamplification with internal primers was used to improve the quality and the quantity of amplified DNA, allowing for post-PCR analysis such as sequence identification of the viral strain. With this procedure, multiple samples could be analyzed in four working days by a single trained individual. The nominal sensitivity of detection of the procedure was 0.15 TCID50 (50% tissue culture infective dose) per 0.62 g of tissue with a test virus. The direct RNA isolation protocol avoided pitfalls associated with whole-virus purification procedures by replacing virus precipitation steps involving polyethylene glycol and Procipitate with phenol extraction. The method is straightforward and reliable. We successfully used this procedure to detect naturally occurring HAV in clams involved in a gastroenteritis outbreak, as well as in cilantro artificially contaminated with a test virus. PMID- 11858195 TI - Migration of a water pulse through fractured porous media. AB - Contaminant transport from waste-disposal sites is strongly affected by the presence of fractures and the degree of fracture matrix interaction. Characterization of potential contaminant plumes at such sites is difficult, both experimentally and numerically. Simulations of water flow through fractured rock were performed to examine the penetration depth of a large pulse of water entering such a system. Construction water traced with lithium bromide was released during the excavation of a tunnel at Yucca Mountain, Nevada, which is located in an unsaturated fractured tuff formation. Modeling of construction water migration is qualitatively compared with bromide-to-chloride ratio (Br/Cl) data for pore-water salts extracted from drillcores. The influences of local heterogeneities in the fracture network and variations in hydrogeologic parameters were examined by sensitivity analyses and Monte Carlo simulations. The simulation results are qualitatively consistent with the observed Br/Cl signals, although these data may only indicate a minimum penetration depth, and water may have migrated farther through the fracture network. PMID- 11858196 TI - Laboratory sand column study of encapsulated buffer release for potential in situ pH control. AB - Encapsulation technology is being investigated as a method for controlling pH in situ at contaminated groundwater sites where pH may limit remediation of organic contaminants. This study examined the effectiveness of using KH2PO4 buffer encapsulated in a pH-sensitive coating to neutralize pH in laboratory sand columns (1.5-1) under a simulated groundwater flow rate and characterized the pattern of capsule release in the flow-through system. Denitrification was used in the columns to increase the pH of the pore water. Each of three columns was equipped with three miniature mesh wells to allow contact of the buffer with column pore water, but capsules (15 g) were inserted into only one column (amended). The two other columns served as amendment (no buffer) and abiotic (no denitrification) controls. Oxidation-reduction potential, dissolved organic and inorganic carbon, NH4+, NO3- +NO2-, PO(4)3-, and pH were measured in the influent, two side ports, and effluent of the columns over time. Near complete conversion of 80 mg N/1 of nitrate and 152 mg/l of ethanol per day resulted in a mean pH increase from 6.2 to 8.2 in the amendment control column. The amended column maintained the target pH of 7.0 +/- 0.2 for 4 weeks until the capsules began to be depleted, after which time the pH slowly started to increase. The capsules exhibited pulses of buffer release, and were effectively dissolved after 7.5 weeks of operation. Base-neutralizing capacity contributed by the encapsulated buffer over the entire study period, calculated as cation equivalents, was 120 mM compared to 8 mM without buffer. This study demonstrates the potential for this technology to mediate pH changes and provides the framework for future studies in the laboratory and in the field, in which pH is controlled in order to enhance organic contaminant remediation by pH-sensitive systems. PMID- 11858197 TI - Honoring our heroes 2001. IAPAC's 2001 Jonathan Mann Health Human Rights Awards. PMID- 11858198 TI - The latest word on lipodystrophy and mitochondrial toxicity (and old words that still resound). PMID- 11858199 TI - 'Focus on our human bond'. PMID- 11858200 TI - Clinton's war on HIV/AIDS. PMID- 11858201 TI - Modeling coagulation kinetics incorporating fractal theories: comparison with observed data. AB - There are currently four possible approaches in modeling coagulation kinetics: the traditional Euclidean rectilinear; the Euclidean curvilinear; the fractal rectilinear; and the fractal curvilinear. The fractal model includes the Euclidean case as a subset. The primary purpose of this research is to investigate which of the rectilinear models among these best predicts the evolution of experimental observed particle size distribution (PSD). Using a fractal rectilinear model previously developed by the authors, model predictions were compared with a series of observed PSD data obtained from estuarine sediment particles in a 2m settling column, where the average velocity gradient (G) was 20 or 40s(-1). Nonlinear parameter estimation was performed to estimate two free parameters for the fractal model (the fractal dimension, DF, and the collision efficiency factor, a), and one free parameter (the collision efficiency factor, alpha) for the Euclidean model. Compared with the observed PSD, the simulation showed that the fractal rectilinear model was best, and that this model fit better for the larger size particles. The estimated DF was between 2.6 and 3.0. The research demonstrated that the alpha's have multiple values for the same observed data, depending on the coagulation model used. This finding is significant because a is currently used as a single value based on the conventional Euclidean rectilinear model. PMID- 11858202 TI - Childhood epilepsy in relation to mental handicap and behavioural disorders. AB - Epidemiological studies indicate that there is a high rate of mental retardation and behavioural problems in children with epilepsy. In some cases both the epilepsy and the mental retardation will have a common cause, such as a metabolic disorder or brain trauma. However, in other children, the epilepsy itself may cause either temporary or permanent learning problems. When permanent learning disability can be prevented it is important to treat the epilepsy early and effectively. Children with specific learning difficulties and memory problems can benefit greatly from appropriate management. There are many causes of behavioural disturbance in children with epilepsy. These causes include the epilepsy itself, treatment of the epilepsy, reactions to the epilepsy, associated brain damage/dysfunction and causes that are equally applicable to children who do not have epilepsy. Identifying the cause or causes in each child allows rational management to be provided. Antiepileptic treatment with medication or surgery can either improve the situation or make matters worse. The treatment should be tailored to the needs of the individual child. If surgery is required, there is a strong argument for performing this early in life, both to allow the greatest opportunity for brain plasticity and also to allow the child full benefit from the important developmental and educational years, without the problems that can be associated with the epilepsy. Skilled management of children with epilepsy who have mental retardation and/or behavioural problems can be very rewarding both for the family and for the professionals involved. PMID- 11858203 TI - Effects of instrumentation, irrigation and dressing with calcium hydroxide on infection in pulpless teeth with periapical bone lesions. AB - AIM: The aim of this study was to evaluate the fate of microorganisms in root canals of teeth with infected pulps and periapical bone lesions with and without the use of calcium hydroxide medication. METHODOLOGY: Endodontic samples were cultured and microorganisms were counted and identified in 43 teeth before (sample 1) and after (sample 2) treatment during the first visit and before (sample 3) and after (sample 4) treatment during the second visit. In the first visit teeth were instrumented and half of the teeth were filled with a thick slurry of calcium hydroxide in sterile saline. The other teeth were obturated with gutta-percha and AH-2 6 sealer. After 4 weeks the teeth with calcium hydroxide were accessed again and after microbiological sampling they were obturated with gutta-percha and AH-26 sealer. RESULTS: The mean total colony forming unit (CFU) counts of positive samples dropped significantly as a result of canal preparation during the first visit from 1.0 x 10(6) to 1.8 x 10(3) (between samples 1 and 2) but increased to 9.3 x 10(3) in the period between the two visits (sample 2 and 3). There was no difference in mean total CFU counts of positive samples between the end of the first (sample 2) and the end of the second visit (sample 4). The most frequently isolated species were Prevotella intermedia, Capnocytophaga spp.. Actinomyces odontolyticus. Propionibacterium acnes and Peptostreptococcus micros. CONCLUSIONS: Although a calcium hydroxide paste was placed in the prepared canals, the number of positive canals had increased in the period between visits. However, the number of microorganisms had only increased to 0.93% of the original number of CFU (sample 1). It is concluded that a calcium hydroxide and sterile saline slurry limits but does not totally prevent regrowth of endodontic bacteria. PMID- 11858204 TI - Influence of rotational speed, torque and operator proficiency on failure of Greater Taper files. AB - AIM: The purpose of this study was to evaluate the influence of rotational speed. torque, and operator experience on the incidence of locking, deformation, and separation of instruments when using a specific Ni-Ti rotary instrumentation technique in extracted human teeth. METHODOLOGY: Greater Taper Ni-Ti rotary instruments (GT) were used in a crown-down technique. In one group (rotational speed evaluation) of canals (n = 300) speeds of 150, 2 50 and 350 r.p.m. (subgroups 1, 2 and 3) were used. Each one of the subgroups included 100 canals. In a second group (evaluation of torque) (n = 300) torque was set at 20, 30 and 55 Ncm (subgroups 4, 5 and 6). In the third group (evaluation of operator proficiency) (n = 300 three operators with varying experience (subgroups 7, 8 and 9) were also compared. Each subgroup included the use of 10 sets of GT rotary instruments and 100 canals of extracted human molars. Each set of instruments was used in up to 10 canals and sterilized before each case. NaOCl 2.5% was used as an irrigant. The number of locked, deformed, and separated instruments was recorded for each group. Statistical analysis was carried out with pairwise comparisons using Fisher's exact tests for each of the failure type. RESULTS: When the influence of rotational speed was evaluated, instrument deformation and separation did not occur in subgroups 1 (150) r.p.m.), 2 (250 r.p.m.), and 3 (350) r.p.m.). Instrument locking occurred in subgroup 3 only. Statistical analysis demonstrated a significant difference between the 150 and 350 r.p.m. groups and between the 250 and 350 r.p.m. groups with respect to instrument locking. In torque evaluation, neither separation, deformation nor locking occurred during the use of the instruments, at 150 r.p.m., and at the different torque values. When the operators were compared, although two instruments were separated in canals prepared by the least experienced operator. Fisher's exact tests did not demonstrate a significant difference between the three subgroups. Instrument locking, deformation, and separation did not occur with the most experienced operator. None of the instruments separated with the trained operator. CONCLUSIONS: Preclinical training in the use of the GT rotary instruments when used with a crown-down technique at 150 r.p.m. was crucial in avoiding instrument separation and reducing the incidence of instrument locking and deformation. PMID- 11858205 TI - Growth rate of Ulva rigida in different Mediterranean eutrophicated sites. AB - The growth of juvenile populations of Ulva rigida C. Agardh was measured by means of immersion in in situ cages against environmental parameters (temperature, incident light, salinity, dissolved inorganic nitrogen and dissolved reactive phosphorus) in four different eutrophicated southern sites: Channel of the Thau lagoon (France), Lido, Sacca Sessola and Fusina stations (Venice lagoon, Italy). The growth curves as a function of temperature showed that, in all cases, the maximal temperature for Ulva growth was 17 degrees C (limitation in growth below 7 degrees C and above around 25 degrees C). The growth analysis of these four sites showed seasonal differences. In the least eutrophicated and calmest Lido station, grazing and dissolved reactive phosphorus (seven times lower at Lido than at Thau) played a key role. At Thau and Fusina, which are eutrophicated and turbid environments, the incident light had a strong impact on growth. Sacca Sessola, with an intermediate position between the above two mentioned situations, showed the highest growth rate. The values and relative growth rate (RGR) curves of the Mediterranean and open-sea northern sites are discussed. In particular, the temperature defines the type of growth curve (unimodal or bimodal) and the incident light is responsible for the low Mediterranean RGR values (<10% day(-1)). PMID- 11858207 TI - Effect of pH on hydrogen production from glucose by a mixed culture. AB - The effect of pH on the conversion of glucose to hydrogen by a mixed culture of fermentative bacteria was evaluated. At 36 degrees C, six hours hydraulic retention, over 90% of glucose was degraded at pH ranging 4.0-7.0, producing biogas and an effluent comprising mostly fatty acids. At the optimal pH of 5.5, the biogas comprised 64 +/- 2% of hydrogen with a yield of 2.1 +/- 0.1 mol-H2/mol glucose and a specific production rate of 4.6 +/- 0.4 l-H2/(g-VSS day). The effluent was composed of acetate (15.3-34.1%) and butyrate (31.2-45.6%), plus smaller quantities of other volatile fatty acids and alcohols. The diversity of microbial communities increased with pH, based on 16S rDNA analysis by denaturing gradient gel electrophoresis (DGGE). PMID- 11858206 TI - Invertase and urease activities in the carotenogenic yeast Xanthophyllomyces dendrorhous (formerly Phaffia rhodozyma). AB - Invertase and urease are enzyme entities highly associated with the cells of the astaxanthin-producer yeast Xanthophyllomyces dendrorhous (Phaffia rhodozyma) during any stage of its cell growth cycle. In this study cellobiose was a more efficient carbon source than sucrose or its hexose counterparts for invertase expression. Extensive ultrasonication or abrasion with glass pearls were required in order to promote enzyme release. In contrast to the yeast whose growth declines above 27 degrees C, the released enzymes displayed a higher optimum temperature range when assayed in vitro. Isoforms from both enzymes could be resolved either by FPLC on DEAE-Sepharose or by an affinity approach on immobilized Concanavalin. The zymogram for invertase showed a pI somewhat less acidic than that of the similar enzyme from S. cerevisiae. PMID- 11858208 TI - Responding to inpatient violence at a psychiatric hospital of special security: a pilot project. AB - Within secure psychiatric hospitals, staff have to manage many difficult and challenging situations. Crichton (1997) suggested that when staff perceive a greater degree of responsibility on patients for their actions there is a particularly morally censorious response. The aim of this pilot study was to discover if this association, discovered using a hypothetical scenario, was also present in how nursing staff respond to real violent patient behaviour. Over a five week period all episodes of inter-personal violence in a hospital with special security were identified and those involved interviewed. Thirty-one episodes of inter-personal violence were identified. A disproportionate number were caused by female patients and patients detained under civil sections of mental health legislation. A personality disorder diagnosis and the staff belief that mental disorder did not reduce the individual patient's blame for the incident were associated with the response of a restrictive sanction (both cases p < 0.01). PMID- 11858209 TI - The sexual profile of rapists in Singapore. AB - To our knowledge, this is the first paper that examines the sexual profile of rapists in Singapore. A semi-structured interview based on a questionnaire about sexual habits was conducted on 62 convicted rapists and a control group of 63 prisoners comprising 32 convicted for violent non-sexual offences and 31 for non violent non-sexual offences. Significantly more rapists masturbated at least once a month and were exposed to pornography within six months before the offence when compared to non-violent controls. Violent controls did not differ significantly from rapists in any aspect of the sexual history. The greater similarity between rapists and violent controls lends support to the concept of rape as a violent rather than sexual offence. A subgroup of 17 'hypersexual' rapists were identified who were either having sex or masturbating very frequently but had still resorted to rape. Compared to the other 46 rapists, the 'hypersexual' rapists were more likely to have fantasised predominantly about rape or bondage and to cite domination, aggression or hostility as reasons for committing rape, suggesting an even stronger element of power and aggression than in their less sexually active counterparts. PMID- 11858210 TI - Evaluation of factors to convert absorbed dose calibrations from graphite to water for the NPL high-energy photon calibration service. AB - The National Physical Laboratory (NPL) provides a high-energy photon calibration service using 4-19 MV x-rays and 60Co gamma-radiation for secondary standard dosemeters in terms of absorbed dose to water. The primary standard used for this service is a graphite calorimeter and so absorbed dose calibrations must be converted from graphite to water. The conversion factors currently in use were determined prior to the launch of this service in 1988. Since then, it has been found that the differences in inherent filtration between the NPL LINAC and typical clinical machines are large enough to affect absorbed dose calibrations and, since 1992, calibrations have been performed in heavily filtered qualities. The conversion factors for heavily filtered qualities were determined by interpolation and extrapolation of lightly filtered results as a function of tissue phantom ratio 20,10 (TPR20,10). This paper aims to evaluate these factors for all mega-voltage photon energies provided by the NPL LINAC for both lightly and heavily filtered qualities and for 60Co y-radiation in two ways. The first method involves the use of the photon fluence-scaling theorem. This states that if two blocks of different material are irradiated by the same photon beam, and if all dimensions are scaled in the inverse ratio of the electron densities of the two media, then, assuming that all photon interactions occur by Compton scatter the photon attenuation and scatter factors at corresponding scaled points of measurement in the phantom will be identical. The second method involves making in-phantom measurements of chamber response at a constant target-chamber distance. Monte Carlo techniques are then used to determine the corresponding dose to the medium in order to determine the chamber calibration factor directly. Values of the ratio of absorbed dose calibration factors in water and in graphite determined in these two ways agree with each other to within 0.2% (1sigma uncertainty). The best fit to both sets of results agrees with values determined in previous work to within 0.3% (1sigma uncertainty). It is found that the conversion factor is not sensitive to beam filtration. PMID- 11858211 TI - Underestimation of access flow by ultrasound dilution flow measurements. AB - For hemodialysis access surveillance, flow measurements are increasingly considered important because they identify accesses at risk of thrombosis. Usually these flow measurements are performed with the ultrasound dilution technique. In a previous patient study it was observed that the resulting flow values were systematically low as compared to magnetic resonance flow measurements, but a satisfactory explanation was lacking. In the present study, we will demonstrate by hemodynamic calculations and in vitro experiments that this discrepancy can be explained by a temporary reduction of the access flow rate, caused by the reversed needle configuration during ultrasound dilution flow measurements. In this configuration. blood is injected retrogressively at one needle and flow between the needles is increased, causing an increased dissipation of energy. The proposed explanation is subsequently confirmed in a patient with a loop graft, by measuring the blood velocity by Doppler ultrasound as a function of reversed dialyzer flow rate. Apart from the ultrasound dilution technique, these findings are applicable to other recently proposed methods for measuring access flow that employ the reversed needle configuration. PMID- 11858212 TI - Letting the Gini out of the bottle? Challenges facing the relative income hypothesis. AB - The relative income hypothesis interprets statistical associations between income inequality and average health status at the population level, as evidence that income inequality has a deleterious psychosocial effect on individual health. An alternative explanation is that these, population-level associations, are statistical artefacts of curvilinear, individual-level relationships between income and health. Indeed, provided the cost-benefit ratio of health-enhancing goods and services vary, the law of diminishing returns should produce curvilinear, asymptotic relationships between income and health at the individual level, which create ('artefactual') associations between income inequality and health at the population level. However, proponents of the relative income hypothesis have argued that these relationships are unlikely to be responsible for the associations observed between income inequality and average health status amongst high-income populations. In these populations, the individual-level relationships between income and health would be nearer their asymptotes, where a shallower slope should ensure that income inequality has little (if any) 'artefactual' effect on average health status. Yet this argument was based on analyses of population-level data which underestimated the slope and curvilinearity of underlying, individual-level relationships between income and health. It is therefore likely that (at least some part of) the population-level associations between income inequality and average health status (amongst low-, middle- and high-income populations) are 'artefacts' of curvilinear, individual level relationships between income and health. Nevertheless, it is also possible that income inequality is somehow (partly or wholly) responsible for the curvilinear nature of individual-level relationships between income and health. Likewise, it is possible that income inequality alters the height, slope and/or curvilinearity of these relationships in such a way that income inequality has an independent effect on individual health. In either instance, the 'artefactual' effect of curvilinear relationships between income and health at the individual level would simply reflect the mechanism underlying the relative income hypothesis. PMID- 11858213 TI - Bronchodilator activity of Mikania glomerata Sprengel on human bronchi and guinea pig trachea. AB - The effects of aqueous extracts and hydro-alcoholic extract (HAE), and of a dichloromethane fraction (MG1) obtained from the HAE of Mikania glomerata leaves on isolated respiratory and vascular smooth muscle have been investigated. Aqueousextracts and HAE induced a significant inhibition on the histamine contractions on the isolated guinea-pig trachea. HAE extract induced a concentration-dependent relaxation on guinea-pig trachea pre-contracted with histamine (IC50 0.34 (0.29-0.39) mg mL(-1)), acetylcholine (IC50 0.72 (0.67-0.77) mg mL(-1)) or K+ (IC50 1.41 (1.18-1.64) mg mL(-1)) and on isolated human bronchi precontracted with K+ (IC50 0.34 (0.26-0.42) mg mL(-1)). The dichloromethane fraction induced a concentration dependent relaxation in guinea-pig trachea precontracted with K+ (IC50 0.017 (0.012-0.022) mg mL(-1)). The dichloromethane fraction had also a small vasodilator effect on the isolated mesenteric vascular bed and on the isolated rat aorta, and a significant reduction of the oedema induced by subplantar injections of Bothropsjararaca venom in mice. When tested on plasmid DNA, MG1 did not damage the DNA. Chromatographic analysis showed the presence of 11.4% w/w coumarin in MG1. The results supported the indication of M. glomerata products for the treatment of respiratory diseases where bronchoconstriction is present. PMID- 11858214 TI - Effects of adenosine agonists on consumptive behaviour and body temperature. AB - This study was designed to determine the effects of the A1-receptor selective agonist N6-cyclopentyladenosine (CPA), and the A2-selective agonist, 2-p-(2 carboxyethyl)-phenethylamino-5'-N-ethylcarboxamidoadenosine-hydrochloride (CGS 21680) on consumptive behaviour and body temperature in rats in relation to the non-selective A1/A2 adenosine agonist, N-ethylcarboxamidoadenosine (NECA), and to morphine. It was shown that two subcutaneous injections of 0.1 and 0.3 mg kg(-1) CPA caused a similar decrease in food consumption to NECA (2 x 0.03 mg kg(-1)) and morphine (2 x 10 mg kg(-1)). However, two doses of 0.03 mg kg(-1) CPA and 0.1 and 0.3 mg kg(-1)CGS-21680 enhanced feeding. These effects were not directly correlated to faecal output at all doses of the selective agonists, as NECA and morphine induced constipation. The doses of CPA and 0.1 and 0.3 mg kg(-1) of CGS 21680 enhanced water consumption, as did NECA, but not morphine. The stimulation of drinking by CPA was not absolutely associated with diuresis. Instead, urine output was reduced by 0.03 and 0.1 mg kg(-1) and increased by 0.3 mg kg(-1). CGS 21680 at 0.1 and 0.3 mg kg(-1) and NECA also induced diuresis, which was opposite to the effect of morphine. CPA and CGS-21680 both caused significant dose dependent decreases in body temperature after the two-injection treatment, but their effects were significantly less after 36 h when four doses had been administered. The study indicates that highly selective A1 and A2A adenosine agonists might have the ability to interfere with consumptive behaviour, induce constipation, affect renal function and to lower body temperature. PMID- 11858215 TI - Identification of indolyl-3-acryloylglycine in the urine of people with autism. AB - HPLC analysis of the urine of autistic subjects indicated the presence of an unidentified component in greatly increased concentrations. We have reported the isolation of this component by HPLC and its identification. Mass spectrometry, NMR and UV spectroscopy identified the peak as corresponding to indolyl-3 acryloylglycine (IAG, 3), and this has been confirmed by an independent synthesis. PMID- 11858216 TI - Bilateral connation of permanent mandibular incisors: a case report. AB - Connated teeth are the consequences of developmental anomalies leading to the eruption of joined elements. According to current definitions, gemination occurs when one tooth bud tries to divide, while fusion occurs if two buds unite. Clinical experience shows, however, that diagnosis can be complicated due to superimposed anomalies. This report describes a unique case of bilateral fusion of permanent mandibular incisors and discusses the differential diagnosis and possible future treatment options anticipated for this particular case. PMID- 11858218 TI - Bone loss and inhaled glucocorticoids. PMID- 11858217 TI - Biological variation in sweat sodium chloride conductivity. AB - BACKGROUND: Sweat conductivity, which is equivalent to sweat NaCl concentration, is used as a screening test to identify possible cystic fibrosis (CF) patients. No data exist on the biological variation of this variable and the influence it may have on the interpretation of sweat testing. The aim of this study was to determine the components of biological variation for sweat sodium chloride conductivity and to apply biological variation parameters in the interpretation of sweat conductivity. METHODS: Sweat conductivity was determined once a week for 5 consecutive weeks on 15 healthy volunteers, 20 healthy infants and 20 known CF patients. RESULTS: The analytical coefficient of variation (CV(A)) was 1.15% for the high-level control material, with a value of 123 mmol/L, and 1.32% for the normal-level control material with a value of 40 mmoL/L. The within-subject (CV) and between-subject (CV(G)) biological variations were 12.0% and 30.0%, respectively, for healthy controls; 18% and 20% for healthy infants; and 7.3% and 6.5% for CF patients, respectively. Using the CV(A), CV(G) and CV(I), the 95% reference ranges were determined for the above-mentioned three groups. The calculated 95% ranges for the healthy babies and CF patients were 18-60 mmoL/L and 96-144 mmoL/L. CONCLUSIONS: Our data support a decision level of > 60 mmoL/L for confirmatory CF testing. A lower decision level will result in an unacceptable high rate of unnecessary confirmation testing. PMID- 11858219 TI - Bone loss and inhaled glucocorticoids. PMID- 11858220 TI - Urinary tract infections and a multidrug-resistant Escherichia coli clonal group. PMID- 11858221 TI - Urinary tract infections and a multidrug-resistant Escherichia coli clonal group. PMID- 11858222 TI - B-cell deficiency and type 1 diabetes. PMID- 11858223 TI - The acts of terrorism. PMID- 11858226 TI - Dichotic listening in patients with splenial and nonsplenial callosal lesions. AB - The authors found splenial lesions to be associated with left ear suppression in dichotic listening of consonant-vowel syllables. This was found in both a rapid presentation dichotic monitoring task and a standard dichotic listening task, ruling out attentional limitations in the processing of high stimulus loads as a confounding factor. Moreover, directed attention to the left ear did not improve left ear target detection in the patients, independent of callosal lesion location. The authors' data may indicate that auditory callosal fibers pass through the splenium more posterior than previously thought. However, further studies should investigate whether callosal fibers between primary and secondary auditory cortices, or between higher level multimodal cortices, are vital for the detection of left ear targets in dichotic listening. PMID- 11858225 TI - The role of the African-American physician in reducing traffic-related injury and death among African Americans: consensus report of the National Medical Association. AB - ISSUE: Traffic-related injuries and fatalities disproportionately affect the African American community. These high rates of traffic-related death and injury among African Americans manifest in multiple areas of traffic safety, including: Failure to use seat belts and child restraints. High incidence of alcohol impaired driving. Failure to follow child passenger and seat belt safety laws and recommendations. High rates of pedestrian accidents, ofen brought on by impairments of drivers and/or pedestrians. Research indicates that national public information campaigns, with general messages only slightly modified for African American audiences, have not been culturally appropriate or effective in changing traffic safety behavior. In addition, traditional distribution mechanisms for these messages have not effectively reached the target population. Evidence suggests that in the African American community, there is a pervasive lack of knowledge of the devastating impact of traffic-related accidents on the overall health status of the community. This lack of information has resulted in a tragic cycle, in which parents fail to model safe operation of motor vehicles, and generation after generation copy this behavior, increasing the community's vulnerability to serious injuries and untimely deaths. This trend toward improper traffic safety habits among African Americans persists despite federal, state and local laws to enforce and promote sound traffic safety practices. OBJECTIVE: To study the existence of disparities in traffic-related injury and death among African Americans and to determine what kinds of traffic safety messages and campaigns will be effective in encouraging African Americans to respond to safety laws in sufficient numbers to reduce the disproportionately high rate of injury and death. Traffic safety issues were examined to effectively recommend policy, address barriers, best practices, and intervention strategies for the National Medical Association, its physician members, their patients, and their communities. CONSENSUS PROCESS: A literature review, driven by research instruments from numerous organizations included reports and materials from the National Highway Traffic Safety Administration (NHTSA), American Academy of Pediatrics, National Committee for Injury Prevention and Control, U.S. Centers for Disease Control and Prevention (CDC), Mothers Against Drunk Driving (MADD), and the National SAFE KIDS Campaign. Both the Meharry Medical College report, Achieving a Credible Health and Safety Approach to Increasing Seat Belt Use Among African-Americans, and the U.S. Department of Transportation's Blue Ribbon Panel to Increase Seat Belt Use Among African Americans: A Report to the Nation, provided substantial background for the panel. More than 60 pieces of traffic safety literature have been examined to date. Based on the literature review, a short list of the most relevant issues affecting African Americans and traffic safety was devised. It includes: The disproportionately high rate of traffic related injury and death among African Americans. The cost in health, monetary costs and other associated costs of traffic safety accidents and injuries. The number of traffic-related injuries and deaths that could be prevented if more African Americans observed good traffic safety practices. Barriers to practicing good traffic safety habits among African Americans. Failure of laws and public information campaigns to influence improved traffic safety practices among African Americans sufficient to reduce disparities in traffic-related injury and death. In July 2001, NMA convened a consensus panel of experts in St. Thomas, U.S. Virgin Islands, to review a briefing document summarizing the most salient traffic safety issues among African Americans. The panel elaborated on key issues, including existing policy and standards for the use of child restraint devices to secure infants and toddlers, existing data regarding disparities in traffic-related injury and death among African Americans, and the cultural, age and developmental appropriateness of existing safety campaigns. SUMMARY: Public information campaigns have successfully improved traffic safety practices among the general public but in large part have been unsuccessful among minority populations-including African Americans. This may be due to: A failure to use techniques and messages that are culturally sensitive to African Americans. Campaigns that have targeted geographic and social centers where African Americans are not broadly present. Lack of awareness of the disproportionate effect motor vehicle crashes are having on African Americans. Scientifically based, culturally appropriate intervention strategies need to be devised and implemented by African American institutions and organizations to improve traffic safety practices and reduce the high rate of traffic-related injury and deaths among African Americans. PMID- 11858227 TI - Characterization of genes encoding multi-domain proteins in the genome of the filamentous nitrogen-fixing Cyanobacterium anabaena sp. strain PCC 7120. AB - Computational analysis of gene structures in the genome of Anabaena sp. PCC 7120 revealed the presence of a large number of genes encoding proteins with multiple functional domains. This was most evident in the genes for signal transduction pathway and the related systems. Comparison of the putative amino acid sequences of the gene products with those in the Pfam database indicated that and PAS domains which may be involved in signal recognition were extremely abundant in Anabaena: 87 GAF domains in 62 ORFs and 140 PAS domains in 59 ORFs. As for the two-component signal transduction system, 73, 53, and 77 genes for simple sensory His kinases, hybrid His kinases and simple response regulators, respectively, many of which contained additional domains of diverse functions, were presumptively assigned. A total of 52 ORFs encoding putative Hanks-type Ser/Thr protein kinases with various domains such as WD-repeat, GAF and His kinase domains, as well as genes for presumptive protein phosphatases, were also identified. In addition, genes for putative transcription factors and for proteins in the cAMP signal transduction system harbored complex gene structures with multiple domains. PMID- 11858228 TI - Application of pharmacoeconomics to formulary decision making in managed care organizations. AB - OBJECTIVE: To discuss the apparent value of incorporating pharmacoeconomic studies into pharmacy and therapeutic committee decision making; current internal and external barriers to the use of pharmacoeconomic studies; and possible solutions to the problems. STUDY DESIGN: Literature review. RESULTS: The formulary system assists healthcare providers in the evaluation, appraisal, and selection of drugs. Unfortunately, managed care organizations usually evaluate drugs exclusively on clinical efficacy, safety, and daily acquisition cost without considering overall cost effectiveness. Factors that have been impeding the use of pharmacoeconomic data include departmental budgetary constraints, tardy publications, limited reliability of available studies, and a lack of knowledge required to evaluate such studies. CONCLUSIONS: To remain competitive, managed care organizations need to incorporate pharmacoeconomic consideration into their formulary decision-making process. Performing an institutionwide economic evaluation; conducting pharmacoeconomic studies earlier, perhaps along with clinical trials; using decision analysis; developing standardized guidelines; and increasing education can help overcome current barriers. PMID- 11858229 TI - Does the school nurse have a role to play in school discipline? AB - Student discipline has become a subject of increasing concern at local, state, and national levels. This article is a discussion of current federal, state, and local school district legal requirements for student discipline in public schools with examples that illustrate several issues that could or should involve a school nurse. A brief history of the legal developments in school discipline includes key U.S. Supreme Court decisions and Acts of Congress. School district options in policy development and disciplinary procedures are discussed. Some of the discipline incidents include issues of nurse-patient confidentiality. PMID- 11858230 TI - Asian domains of four major genotypes of JC virus, Af2, B1-b, CY and SC. AB - JC virus (JCV) strains worldwide can be classified into various genotypes based on DNA sequence variations. To define the domains of the four major JCV genotypes in Asia, we collected urine samples at six unstudied sites: three in southeastern Asia, two in the central highlands and one in central Asia. DNA was extracted from urine samples, and used to amplify a 610-bp region of the viral genome. For each geographical site, we determined 16 to 31 sequences, from which a phylogenetic tree was constructed to unambiguously classify detected JCV isolates into distinct genotypes. From JCV genotype profiles at the sites studied here and elsewhere, the following conclusions were drawn. Although Af2 is the major genotype in Africa, this genotype also occurs in western and central Asia. B1-b mainly occurs in western and central Asia, including the central highlands. CY occurs in northeastern Asia with the southern boundary between China and southeast Asian countries. Although SC predominates in southeastern Asia, it also occurs in northern and central Asia at lower frequencies. In addition, a few minor JCV genotypes (B1-a, B2 and B3) occur at many sites. We discuss here the anthropological and medical significance of the present findings. PMID- 11858231 TI - Genetic diversity of Wheat dwarf virus. AB - The complete sequence of a new wheat-infecting isolate of Wheat dwarf virus from Sweden (WDV-[Enk1]) was determined, as well as a 726-nt region covering part of rep and the long intergenic region (LIR) of six other wheat-infecting Swedish isolates and a barley-infecting isolate from Hungary (WDV-Bar[HU]). Analyses including these and previously published sequences showed that the wheat infecting isolates of WDV displayed less than 3% of divergence. Most of the nucleotide changes were silent and the largest variation was detected in LIR. In contrast, the barley-infecting isolate was clearly different with 16% of sequence divergence compared to the wheat isolates. The WDV isolates infecting barley or wheat seem to represent two differentiated strains. PMID- 11858232 TI - Distal colonic motor activity in four subgroups of patients with irritable bowel syndrome. AB - With the aim of improving end organ treatment, we describe a new system of classifying irritable bowel syndrome (IBS) according to clinical features into four groups, spastic colon syndrome (SCS), functional diarrhea (FD), diarrhea predominant spastic colon syndrome (DPSCS), and midgut dysmotility (MGD). The aim of the study was to investigate fasting and postprandial distal colonic motility in the four groups of patients and to compare the results with normal controls. Distal colonic motility studies were performed in the unprepared colon. 2.5-hr recordings were made from four channels with a standard meal administered at 0.5 hr. The intubated colon was treated as a study segment and data analyzed for study segment activity index (SSAI) and number and mean amplitude of pressure peaks over 30-min epochs. Patients with SCS had significantly higher (P < 0.05) mean amplitude of pressure peaks (60 min, 120 min) and SSAI (120 min) than controls and patients with FD, DPSCS, and MGD. In contrast, patients with FD and DPSCS had significantly (P < 0.05) lower postprandial SSAI than controls and patients with SCS (60 min, 120 min). With the exception of raised postprandial mean amplitude of pressure peaks (120 min), MGD patients had normal distal colonic motility. Division of IBS patients into subgroups has highlighted significant differences in distal colonic motility that provide insights into etiopathogenesis and should assist targeting of current and newly developed therapies, particularly receptor active agents. PMID- 11858233 TI - Is there a one-to-one correlation between gastric emptying of liquids and gastric myoelectrical or motor activity in dogs? AB - The aim of this study was to investigate the correlation of gastric emptying with gastric myoelectrical activity recorded from internal serosal electrodes and with gastric motility measured from strain gauges. The study was performed in eight healthy dogs chronically implanted with four pairs of electrodes and two strain gauges on the gastric serosa and equipped with a duodenal fistula for the assessment of gastric emptying. Each dog was fed four liquid test meals on four separate days with identical calories (320 kcal) and volume (473 ml). A correlation was found between gastric emptying and the energy of contraction in the frequency band of 3.75-7.50 cycles/min during the first 30 min after the meal (r = 0.46, P < 0.05). While none of the parameters of gastric myoelectrical activity was correlated with gastric emptying, two major parameters, percentage of regular gastric slow waves and percentage of slow-wave coupling, were found to be associated with delayed gastric emptying. A significant correlation was found between the frequency of gastric slow waves and that of the contractions in both fasting (r = 0.83, P < 0.001) and fed state (r = 0.70, P < 0.001 at 30 min, r = 0.86, P < 0.001 at 60 min). It was concluded that gastric emptying is correlated with the strength of gastric contractions in a frequency range identical to that of the gastric slow waves, and there is no one-to-one correlation between gastric emptying of liquids and any parameters of gastric myoelectrical activity. However, major parameters of gastric myoelectrical activity are associated with gastric emptying. PMID- 11858235 TI - Abstracts of the American Society of Critical Care Anesthesiologists 14th annual meeting. New Orleans, Louisiana, USA. October 12, 2001. PMID- 11858234 TI - Online regulatory guide errors. PMID- 11858236 TI - Diabetes and coronary heart disease: a time for action. PMID- 11858237 TI - Use of mechanical ventilation challenged in hospice case. PMID- 11858239 TI - Promoting spiritual health is responsibility of nurses. PMID- 11858238 TI - Corporate systems threaten nursing. PMID- 11858240 TI - Debate over use of phenytoin. PMID- 11858243 TI - Brugada syndrome: a case study of aborted sudden cardiac death manifesting as seizures. PMID- 11858242 TI - Diagnosing and treating pacemaker syndrome. PMID- 11858244 TI - Managing delirium in adult intensive care patients. PMID- 11858245 TI - Understanding advanced concepts in atrioventricular block. AB - Not all atrioventricular blocks occur in the atrioventricular node. Understanding the physiological concept behind the various blocks makes recognizing the relevant ECG indications easier. Furthermore, the implications to patients are clear when healthcare providers understand the level of the block and the dangers the block may impose. Until a more accurately descriptive classification system of conduction blocks becomes universally accepted, the current labels will continue to be used. It is imperative that nurses, particularly those caring for cardiac patients, recognize and understand the ECG indications of various degrees of block. PMID- 11858246 TI - Caring for patients with HIV disease in the new millennium. PMID- 11858247 TI - S(VO)2 monitoring. PMID- 11858248 TI - Practice guidelines for prone positioning of acutely ill patients. PMID- 11858249 TI - Procedure boxes save time, save steps, save money. PMID- 11858250 TI - Abstracts of the 31st Annual Meeting of the German Society for Thoracic and Cardiovascular Surgery. February 17-20, 2002. Leipzig, Germany. PMID- 11858251 TI - Induced phenylpropanoid metabolism during suberization and lignification: a comparative analysis. AB - Induction of the biosynthesis of phenylpropanoids was monitored at the enzyme level through measurement of the temporal change in the activity of two marker enzymes of phenylpropanoid metabolism, phenylalanine ammonia-lyase, (PAL, E.C. 4.1.3.5) and 4-coumaryl-CoA ligase (4-CL, E.C. 6.2.1.12) and two marker enzymes for hydroxycinnamyl alcohol biosynthesis, cinnamoyl-CoA:NADP+ oxidoreductase (CCR, E.C. 1.2.1.44) and cinnamyl alcohol dehydrogenase (CAD, E.C. 1.1.1.195) in both suberizing potato (Solanum tuberosum) tubers and lignifying loblolly pine (Pinus taeda) cell cultures. While measurable activities of PAL, 4-CL and CAD increased upon initiation of suberization in potato tubers, that of CCR did not. By contrast, all four enzymes were induced upon initiation of lignification in pine cell cultures. The lack of CCR induction in potato by wound treatment is consistent with the channelling of hydroxycinnamoyl-CoA derivatives away from monolignol formation and toward other hydroxycinnamoyl derivatives such as those that accumulate during suberization. PMID- 11858252 TI - Hardware of the flight experiment "Quail-SK" transport incubator. AB - The transportation of quail eggs in various stages of incubation was used in an experiment at the orbital station MIR by the Slovak astronaut, Bella in February 1999. Device description, diagrams, and experimental results are presented in this paper. PMID- 11858253 TI - Formation of higher plant component microbial community in closed ecological system. AB - Closed ecological systems (CES) place at the disposal of a researcher unique possibilities to study the role of microbial communities in individual components and of the entire system. The microbial community of the higher plant component has been found to form depending on specific conditions of the closed ecosystem: length of time the solution is reused, introduction of intrasystem waste water into the nutrient medium, effect of other component of the system, and system closure in terms of gas exchange. The higher plant component formed its own microbial complex different from that formed prior to closure. The microbial complex of vegetable polyculture is more diverse and stable than the monoculture of wheat. The composition of the components' microflora changed, species diversity decreased, individual species of bacteria and fungi whose numbers were not so great before the closure prevailed. Special attention should be paid to phytopathogenic and conditionally pathogenic species of microorganisms potentially hazardous to man or plants and the least controlled in CES. This situation can endanger creation of CES and make conjectural existence of preplanned components, man, specifically, and consequently, of CES as it is. PMID- 11858254 TI - Interaction peculiarities between microbial cenosis and local immunity of periodontium of humans under extreme conditions. AB - The investigations were directed at finding out the peculiarities of both periodontal microflora and local immunity under extreme conditions (spaceflight, saturation diving, antiorthostatic hypokinesia). The obtained results revealed different kinds of interactions amongst which saturation diving resulted in the most severe decrease of commensale microflora level, decrease of local immunity and appearance of periodontal pathogens. PMID- 11858255 TI - Response of silicon-based linear energy transfer spectrometers: implication for radiation risk assessment in space flights. AB - There is considerable interest in developing silicon-based telescopes because of their compactness and low power requirements. Three such telescopes have been flown on board the Space Shuttle to measure the linear energy transfer spectra of trapped, galactic cosmic ray, and solar energetic particles. Dosimeters based on single silicon detectors have also been flown on the Mir orbital station. A comparison of the absorbed dose and radiation quality factors calculated from these telescopes with that estimated from measurements made with a tissue equivalent proportional counter show differences which need to be fully understood if these telescopes are to be used for astronaut radiation risk assessments. Instrument performance is complicated by a variety of factors. A Monte Carlo-based technique was developed to model the behavior of both single element detectors in a proton beam, and the performance of a two-element, wide angle telescope, in the trapped belt proton field inside the Space Shuttle. The technique is based on: (1) radiation transport intranuclear-evaporation model that takes into account the charge and angular distribution of target fragments, (2) Landau-Vavilov distribution of energy deposition allowing for electron escape, (3) true detector geometry of the telescope, (4) coincidence and discriminator settings, (5) spacecraft shielding geometry, and (6) the external space radiation environment, including albedo protons. The value of such detailed modeling and its implications in astronaut risk assessment is addressed. PMID- 11858256 TI - Runway width effects in the visual approach to landing. AB - The effect of changes in runway width on the perception of glide-slope has been the subject of extensive investigations. Despite considerable research, an explanation of this effect has been elusive. A mathematical model for glide-slope perception was published recently based on the premise that a desirable goal of perception is to form a perception with minimal uncertainty. One of the qualitative predictions of that model was that changes in the aspect ratio of the runway would affect the perceived glide-slope. In this article, the predictions of the model are quantified and are shown to be in close agreement with experimental results in the literature. PMID- 11858258 TI - Clinical hypnosis and anesthesia: an historical review and its clinical implications in today's practice. PMID- 11858257 TI - Greenhouse tomato limited cluster production systems: crop management practices affect yield. AB - Limited-cluster production systems may be a useful strategy to increase crop production and profitability for the greenhouse tomato (Lycopersicon esculentum Mill). In this study, using an ebb-and-flood hydroponics system, we modified plant architecture and spacing and determined the effects on fruit yield and harvest index at two light levels. Single-cluster plants pruned to allow two leaves above the cluster had 25% higher fruit yields than did plants pruned directly above the cluster; this was due to an increase in fruit weight, not fruit number. Both fruit yield and harvest index were greater for all single cluster plants at the higher light level because of increases in both fruit weight and fruit number. Fruit yield for two-cluster plants was 30% to 40% higher than for single-cluster plants, and there was little difference in the dates or length of the harvest period. Fruit yield for three-cluster plants was not significantly different from that of two-cluster plants; moreover, the harvest period was delayed by 5 days. Plant density (5.5, 7.4, 9.2 plants/m2) affected fruit yield/plant, but not fruit yield/unit area. Given the higher costs for materials and labor associated with higher plant densities, a two-cluster crop at 5.5 plants/m2 with two leaves above the cluster was the best of the production system strategies tested. PMID- 11858259 TI - Root elongation against a constant force: experiment with a computerized feedback controlled device. AB - Axial force was applied to the root tip of corn (Zea mays L. cv. Merit) seedlings using a computerized, feedback-controlled mechanical device. The system's feedback capability allowed continuous control of a constant tip load, and the attached displacement transducer provided the time course of root elongation. Loads up to 7.5 g decreased the root elongation rate by 0.13 mm h-1 g-1, but loads 7.5 to 17.5 g decreased the growth rate by only 0.04 mm h-1 g-1. Loads higher than 18 g stopped root elongation completely. Measurement of the cross sectional areas of the root tips indicated that the 18 g load had applied about 0.98 MPa of axial pressure to the root, thereby exceeding the root's ability to respond with increased turgor pressure. Recorded time-lapse images of loaded roots showed that radial thickening (swelling) occurred behind the root cap, whose cross-sectional area increased with tip load. PMID- 11858260 TI - The wounding, amputation and death of Thomas Jonathan "Stonewall" Jackson: some medical and historical insights. PMID- 11858261 TI - An enduring controversy: Henry K. Beecher and curare. PMID- 11858262 TI - Differential ethylene sensitivity of epidermal cells is involved in the establishment of cell pattern in the Arabidopsis root. AB - Root hairs of Arabidopsis roots develop on trichoblasts located over the anticlinal (radial) walls of underlying cortical cells. Non-hair cells, on the other hand, develop on atrichoblasts overlying the periclinal (tangential) walls of cortical cells. Dark-grown wild-type seedlings, which produce little ethylene, are largely root hairless. Exogenous treatment of dark-grown plants with either ethylene or 1-aminocyclopropane-1-carboxylic acid (ACC) restores the development of root hairs in cells overlying the anticlinal cortical cell walls, indicating that cells in this position are more sensitive to ethylene than atrichoblasts. We used mutations in genes that overproduce ethylene (eto1, eto2, eto3 and eto4) to illustrate the positive regulatory role of ethylene. The preferential development of root hairs on epidermal cells overlying the cortical anticlinal cell walls in these mutants also illustrates that trichoblasts are more sensitive to ethylene than atrichoblasts. CTR1 is a negative regulator of the ethylene response and might, therefore, be a candidate regulator of differential sensitivity. CTR1 mRNA is expressed in all cell types in the root, suggesting that its transcriptional pattern alone cannot account for the differential sensitivity of epidermal cells to ethylene. Cellular mapping of wild-type and mutant roots supports previous findings indicating that ethylene acts after, and perhaps independently, of TTG during the establishment of cell fate in the root epidermis. PMID- 11858263 TI - The Cerne 'trembling' hand and the tremulous hand of Worcester. PMID- 11858264 TI - Williams's "The Red wheelbarrow". PMID- 11858265 TI - Some aspects of mortality in three Shropshire parishes in the mid-seventeenth century. PMID- 11858266 TI - Chemotherapy in the elderly. PMID- 11858267 TI - Chemotherapy in the elderly. PMID- 11858268 TI - Profile of the Owen H. Wangensteen Historical Library of Biology and Medicine. PMID- 11858269 TI - Main problems of the Russian Orlan-M space suit utilization for EVAs on the ISS. AB - In the recent years the Russian Orlan-M space suits have been improved as applied to their operational requirements for the ISS. A special attention is paid to enhancement of EVA crew efficiency and safety. The paper considers the main problems regarding specific features of the Russian space suit operation in the ISS, and analyses measures on their solution. In particular, the problems associated with the following are considered: enhancement of the anthropometric range for the EVA crewmembers; use of some US EMU elements and unified NASA equipment elements; Orlan-M operation support in the wide range of the ISS thermal conditions; use of Simplified Aid For Extravehicular activity Rescue (SAFER) designed as a self-rescue device, which will be used for an EVA crewmember return in the event that he (she) breaks away inadvertently from the ISS surface. The paper states the main space suit differences with reference to solution of the above problems. The paper presents briefly the design of space suit arms developed for crewmembers with small anthropometric parameters, as well as peculiarities and test results for the gloves with enhanced thermal protection. Measures on further space suit development with the purpose to improve its performances are considered. PMID- 11858271 TI - Potential and benefits of closed loop ECLS systems on the ISS. AB - To close open loops for long manned missions in space is a big challenge for aeronautic engineers throughout the world. The paper's focus is on the oxygen reclamation from carbon dioxide within a space habitat. A brief description of the function principle of a fixed alkaline electrolyzer, a solid amine carbon dioxide concentrator and a Sabatier reactor is given. By combining these devices to an air revitalization system the technical and economical benefits are explained. Astrium's Air Revitalization System (ARES) as a potential future part of the International Space Station's Environmental Control and Life Support System would close the oxygen loop. The amount of oxygen, needed for an ISS crew of seven astronauts could be provided by ARES. The upload of almost 1500 kg of water annually for oxygen generation through the onboard electrolyzer would be reduced by more than 1000 kg, resulting in savings of more than 30M$ per year. Additionally, the payload capacity of supply flights would be increased by this amount of mass. Further possibilities are addressed to combine ECLS mass flows with those of the power, propulsion and attitude control systems. Such closed loop approaches will contribute to ease long time missions (e. g. Mars, Moon) from a cost and logistic point of view. The hardware realization of Astrium's space-sized operating ARES is shown and test results of continuous and intermittent closed chamber tests are presented. PMID- 11858270 TI - Aquatic modules for bioregenerative life support systems based on the C.E.B.A.S. biotechnology [correction of biotechnilogy]. AB - Most concepts for bioregenerative life support systems are based on edible higher land plants which create some problems with growth and seed generation under space conditions. Animal protein production is mostly neglected because of the tremendous waste management problems with tetrapods under reduced weightlessness. Therefore, the "Closed Equilibrated Biological Aquatic System" (C.E.B.A.S.) was developed which represents an artificial aquatic ecosystem containing aquatic organisms which are adapted at all to "near weightlessness conditions" (fishes Xiphophorus helleri, water snails Biomphalaria glabrata, ammonia oxidizing bacteria and the rootless non-gravitropic edible water plant Ceratophyllum demersum). Basically the C.E.B.A.S. consists of 4 subsystems: a ZOOLOGICAL (correction of ZOOLOGICASL) COMPONENT (animal aquarium), a BOTANICAL COMPONENT (aquatic plant bioreactor), a MICROBIAL COMPONENT (bacteria filter) and an ELECTRONICAL COMPONENT (data acquisition and control unit). Superficially, the function principle appears simple: the plants convert light energy into chemical energy via photosynthesis thus producing biomass and oxygen. The animals and microorganisms use the oxygen for respiration and produce the carbon dioxide which is essential for plant photosynthesis. The ammonia ions excreted by the animals are converted by the bacteria to nitrite and then to nitrate ions which serve as a nitrogen source for the plants. Other essential ions derive from biological degradation of animal waste products and dead organic matter. The C.E.B.A.S. exists in 2 basic versions: the original C.E.B.A.S. with a volume of 150 liters and a self-sustaining standing time of more than 13 month and the so called C.E.B.A.S. MINI MODULE with a volume of about 8.5 liters. In the latter there is no closed food loop by reasons of available space so that animal food has to be provided via an automated feeder. This device was flown already successfully on the STS-89 and STS-90 spaceshuttle missions and the working hypothesis was verified that aquatic organisms are nearly not affected at all by space conditions, i.e. that the plants exhibited biomass production rates identical to the sound controls and that as well the reproductive, and the immune system as the embryonic and ontogenic development of the animals remained undisturbed. Currently the C.E.B.A.S. MINI MODLULE is prepared for a third spaceshuttle flight (STS-107) in spring 2001. Based on the results of the space experiments a series of prototypes of aquatic food production modules for the implementation into BLSS were developed. This paper describes the scientific disposition of the STS-107 experiment and of open and closed aquaculture systems based on another aquatic plant species, the Lemnacean Wolffia arrhiza which is cultured as a vegetable in Southeastern Asia. This plant can be grown in suspension culture and several special bioreactors were developed for this purpose. W. arrhiza reproduces mainly vegetatively by buds but also sexually from time to time and is therefore especially suitable for genetic engineering, too. Therefore it was used, in addition, to optimize the C.E.B.A.S. MINI MODULE to allow experiments with a duration of 4 month in the International Space Station the basic principle of which will be explained. In the context of aquaculture systems for BLSS the continuous replacement of removed fish biomass is an essential demand. Although fish reproduction seems not to be affected in the shortterm space experiments with the C.E.B.A.S. MINI MODULE a functional and reliable hatchery for the production of siblings under reduced weightlessness is connected with some serious problems. Therefore an automated "reproduction module" for the herbivorous fish Tilapia rendalli was developed as a laboratory prototype. It is concluded that aquatic modules of different degrees of complexity can optimize the productivity of BLSS based on higher land plants and that they offer an unique opportunity for the production of animal protein in lunar or planetary bases. PMID- 11858272 TI - The role of cytoskeleton in cell changes under condition of simulated microgravity. AB - Single cells and cell culture are very good model for estimation of primary effects of gravitational changes. It is suggested that cell cytoskeleton plays a key role in mechanisms of adaptation to mechanical influences including gravitational ones. Our results demonstrated that cultured cells of human vascular endothelium (correction of endotheliun) are highly sensitive to hypogravity (clinorotation) and respond by significant decrease of cell proliferative activity. Simultaneously it was noted that the formation of confluent monolayer appeared early in cultures exposed to simulated microgravity due to accelerated cells spreading. Long-term hypogravity (several hours or days) leads to significant changes of cell cytoskeleton revealed as microfilament thinning and their redistribution within cell. Such changes were observed only in monolayer cells and not in cell suspensions. Gravitational forces as known to be modificators of cell adhesive ability and determine their mobility. Hypogravity environment stimulated endothelial cell migration in culture: 24-48 hrs pre exposition to hypogravity significantly increased endothelial cell migration resulting in 2-3-fold acceleration of mechanically injured monolayer repair. Obtained results suggest that the effects of hypogravity on cultured human endothelial cells are, possibly, associated with protein kinase C and/or adenylate cyclase activity and are accompanied by noticeable functional cell changes. PMID- 11858273 TI - Self-sustaining Mars colonies utilizing the North Polar Cap and the Martian atmosphere. AB - A revolutionary new concept for the early establishment of robust, self sustaining Martian colonies is described. The colonies would be located on the North Polar Cap of Mars and utilize readily available water ice and the CO2 Martian atmosphere as raw materials to produce all of the propellants, fuel, air, water, plastics, food, and other supplies needed by the colony. The colonists would live in thermally insulated large, comfortable habitats under the ice surface, fully shielded from cosmic rays. The habitats and supplies would be produced by a compact, lightweight (~4 metric tons) nuclear powered robotic unit termed ALPH (Atomic Liberation of Propellant and Habitat), which would land 2 years before the colonists arrived. Using a compact, lightweight 5 MW (th) nuclear reactor/steam turbine (1 MW(e)) power source and small process units (e.g., H2O electrolyzer, H2 and O2 liquefiers, methanator, plastic polymerizer, food producer, etc.) ALPH would stockpile many hundreds of tons of supplies in melt cavities under the ice, plus insulated habitats, to be in place and ready for use when the colonists landed. With the stockpiled supplies, the colonists would construct and operate rovers and flyers to explore the surface of Mars. ALPH greatly reduces the amount of Earth supplied material needed and enables large permanent colonies on Mars. It also greatly reduces human and mission risks and vastly increases the capability not only for exploration of the surrounding Martian surface, but also the ice cap itself. The North Polar Cap is at the center of the vast ancient ocean that covered much of the Martian Northern Hemisphere. Small, nuclear heated robotic probes would travel deep (1 km or more) inside the ice cap, collecting data on its internal structure, the composition and properties of the ancient Martian atmosphere, and possible evidence of ancient life forms (microfossils, traces of DNA, etc.) that were deposited either by wind or as remnants of the ancient ocean. Details of the ALPH system, which is based on existing technology, are presented. ALPH units could be developed and demonstrated on Earth ice sheets within a few years. An Earth-Mars space transport architecture is described, in which Mars produced propellant and supplies for return journeys to Earth would be lifted with relatively low DeltaV to Mars orbit, and from there transported back to Earth orbit, enabling faster and lower cost trips from Earth to Mars. The exploration capability and quality of life in a mature Martian colony of 500 persons located on the North Polar Cap is outlined. PMID- 11858274 TI - Human interactions during Shuttle/Mir space missions. AB - To improve the interpersonal climate of crewmembers involved with long-duration space missions, it is important to understand the factors affecting their interactions with each other and with members of mission control. This paper will present findings from a recently completed NASA-funded study during the Shuttle/Mir program which evaluated in-group/out-group displacement of negative emotions; changes in tension, cohesion, and leader support over time; and cultural differences. In-flight data were collected from 5 astronauts, 8 cosmonauts, and 42 American and 16 Russian mission control personnel who signed informed consent. Subjects completed a weekly questionnaire that assessed their mood and perception of their work group's interpersonal climate using questions from well-known, standardized measures (Profile of Mood States, Group and Work Environment Scales) and a critical incident log. There was strong evidence for the displacement of tension and dysphoric emotions from crewmembers to mission control personnel and from mission control personnel to management. There was a perceived decrease in commander support during the 2nd half of the missions, and for American crewmembers a novelty effect was found on several subscales during the first few months on-orbit. There were a number of differences between American and Russian responses which suggested that the former were less happy with their interpersonal environment than the latter. Mission control personnel reported more tension and dysphoria than crewmembers, although both groups scored better than other work groups on Earth. Nearly all reported critical incidents came from ground subjects, with Americans and Russians showing important differences in response frequencies. PMID- 11858275 TI - Engineering challenges to the long term operation of the International Space Station. AB - The U.S. Congress has maintained an intense interest in the ISS program since its inception. In the Appropriations Act of 1997, the Senate of the United States included language directing National Aeronautics and Space Administration (NASA) to have the National Research Council (NRC) under take a study that evaluates the engineering challenges posed by extravehicular activity (EVA) requirements, United States and non-United States space launch requirements, the potential need to upgrade or replace equipment and components after Assembly Complete, and the requirement to decommission and disassemble the facility. NASA and the NRC decided the focus should be on the anticipated challenges in the continuous operation and maintenance of the ISS after assembly of the on-orbit facility has been completed. This would encompass the operational years, from late 2004 (if the current schedule holds) to 2020-2025. This executive summary overviews the results of this NRC study. It focuses on the U.S. operation of the ISS after Assembly Complete, including cooperative efforts by the United States and Russia. The paper summarizes the primary findings and recommendations in each of the areas considered during this two-year NRC study. PMID- 11858276 TI - Phase 1 research program overview. AB - The Phase 1 research program was unprecedented in its scope and ambitious in its objectives. The National Aeronautics and Space Administration committed to conducting a multidisciplinary long-duration research program on a platform whose capabilities were not well known, not to mention belonging to another country. For the United States, it provided the first opportunity to conduct research in a long-duration space flight environment since the Skylab program in the 1970's. Multiple technical as well as cultural challenges were successfully overcome through the dedicated efforts of a relatively small cadre of individuals. The program developed processes to successfully plan, train for and execute research in a long-duration environment, with significant differences identified from short-duration space flight science operations. Between August 1994 and June 1998, thousands of kilograms of research hardware was prepared and launched to Mir, and thousands of kilograms of hardware and data products were returned to Earth. More than 150 Principal Investigators from eight countries were involved in the program in seven major research disciplines: Advanced Technology; Earth Sciences; Fundamental Biology; Human Life Sciences; International Space Station Risk Mitigation; Microgravity; and Space Sciences. Approximately 75 long-duration investigations were completed on Mir, with additional investigations performed on the Shuttle flights that docked with Mir. The flight phase included the participation of seven US astronauts and 20 Russian cosmonauts. The successful completion of the Phase 1 research program not only resulted in high quality science return but also in numerous lessons learned to make the ISS experience more productive. The cooperation developed during the program was instrumental in its success. PMID- 11858277 TI - NASA strategic planning for 2000 and beyond. AB - This short paper outlines the United States National Aeronautics and Space Administration's development of its 2000 Strategic Plan. PMID- 11858278 TI - Radical behaviorism and psychology's public: B. F. Skinner in the popular press, 1934-1990. AB - B. F. Skinner is perhaps 2nd only to Freud among the most publicly identifiable psychological figures of the last century. This article reviews the popular press coverage of Skinner between 1934 and 1990 to examine how radical behaviorism was interpreted, portrayed, and received by psychology's public. Reactions to Skinner were often skeptical or condemnatory. It is suggested that some members of the public had difficulty accepting his views because of the disparities between the philosophy of radical behaviorism and the phenomenology of everyday experience. Furthermore, Skinner's status as a psychological expert was inextricably linked to the public's perception of his credibility not only as a scientist but also as a human being. PMID- 11858279 TI - George D. Snell. PMID- 11858281 TI - Hospital charity care policies and the California health care safety net. AB - Estimates of charity care expenditures provide a quantitative measure of hospitals' commitment to expanding access to the medically uninsured in their communities but fail to provide information about the subset of uninsured who benefit or the gaps in public insurance programs' coverage that are filled by these charity care programs. This brief provides an overview of a study, conducted for the California HealthCare Foundation, that gathered information on California hospitals' charity care policies. The objective of the study was to identify the eligibility criteria hospitals use for free care and to understand the gaps in Medi-Cal, California's version of Medicaid, and county indigent program coverage that these providers are capable of filling. PMID- 11858280 TI - [Premature death and longevity of physicians]. AB - The author has divided his work into parts. The first part entitled "Premature Death of Physicians" is dedicated to those who started their research, scientific works and fruitful medical practice but the premature death has stopped their lives and activities. Death causes are presented in ten chapters (groups of causes), i.e. Tuberculosis - Other Lung Diseases - Heart and Vessel Diseases - Septicaemiae - Infectious Diseases in Subgroups: Typhus, Plague, Cholera, Yellow Fever, Diphtheria, Influenza, Malaria, Smallpox, etc. - Encephalopathies and Mental Diseases - Malignant Neoplasms - Noninfectious Unit Diseases - Accidents - Manslaughters - Death Sentences - Suicides - Not Settled Causes of Death. There are in total 283 biographies in the first part. The second part "Longevity of Physicians" is much longer than the first one and contains 509 biographies of doctor, scientists, research workers and practitioners, meritorious in the history of medicine who attained at least 80 years of age. The biographies are arranged in 22 chapters, one for every year from 80 years of age assumed as the beginning of longevity up to 104 years in one of the cases. In each chapter the biographies are arranged in the alphabetical order. In the Epilogue the author presents shortly his conclusions and observations related to the first part and wider commentaries for the second part. PMID- 11858282 TI - Competing visions of the state and social welfare: the Medici Dukes, the Bigallo magistrates, and local hospitals in sixteenth-century Tuscany. AB - In 1542, Florence's Duke Cosimo I established a magistracy to supervise territorial hospitals and consolidate poor relief. Tense relations between the magistracy and these hospitals demonstrate the barriers to bureaucratic centralization in the sixteenth-century state, and underscore the fact that the shift from traditional charity to 'new philanthropy' was as much geographical and cultural as temporal. Tensions between the magistracy and successive Medici Dukes also demonstrate how in negotiations between bureaucrats and local communities territorial rulers could play both sides to advance their personal authority, and could learn from the difficulties of one magistracy how better to design another. PMID- 11858283 TI - The failure of traditional arguments in the vivisection debate. PMID- 11858284 TI - An argument for limited human cloning. PMID- 11858285 TI - Defending abortion: should we treat the body as property? PMID- 11858286 TI - Matters of life and death: inheritance consequences of reproductive technologies. PMID- 11858287 TI - Defining questions: situating issues of power in the formation of a right to health under international law. PMID- 11858288 TI - Doctors' dirty little secrets: the dark side of medical privacy. PMID- 11858289 TI - A universal healthcare system: can it happen in the United States? PMID- 11858290 TI - Community-acquired pneumonia: a case study. AB - Community-acquired pneumonia (CAP) is an infectious disease commonly seen in the primary care environment. CAP is ranked as the sixth leading cause of death in the United States. It affects more than 10 million individuals per year and accounts for 500,000-1,000,000 hospital admissions. Precise diagnosis of CAP continues to be complex. Despite enormous advances in serologic analysis and antimicrobial development, at least 50% of causative pathogens are not identified. This article presents a case report of a white female in her seventies diagnosed with CAP and emphasizes the need for early detection, immunization, and prompt interventions to reduce morbidity and mortality rates. This case brings to light the dilemma practitioners can face when treating high risk populations on an outpatient basis. PMID- 11858291 TI - Back pain: a case study. AB - Musculoskeletal complaints, especially back pain without trauma, are frequent health problems seen by nurse practitioners in community emergency centers and office settings. Back pain can be a symptom of serious health problems. This article presents the case of a Caucasian male in his early sixties who reported sudden onset of back pain after pushing a heavy object. Careful clinical assessment led the nurse practitioner with the collaborating physician to pursue diagnostic tests, which revealed thoracic and abdominal aortic aneurysms. PMID- 11858292 TI - Prescribing patterns of advanced practice nurses: contrasting psychiatric mental health CNS and NP practice. AB - Advanced practice nurses (APNs) play a key role in delivering health care and increasingly include prescriptive authority in their practices. To date, few systematic studies have investigated APN prescribing patterns. The purpose of this investigation was to explore the prescriptive practice of APNs, assess differences in prescriptive patterns by specialty, and determine barriers to prescriptive practice. Fifty-one percent (n = 1352) of 2,651 nurse practitioners (NPs) and psychiatric clinical nurse specialists (CNSs) in Massachusetts responded to a self-administered survey; 59% had incorporated prescriptive authority. Identified barriers included fees, the process of obtaining prescriptive authority, and lack of physician supervision. Significantly more psychiatric CNSs than NPs had difficulty obtaining physician supervisors. Prescriptive authority is integral to advanced practice, but differences in ability to implement this in practice exist among specialties. PMID- 11858293 TI - Hormone therapy and breast cancer. AB - Use or nonuse of hormone therapy (HT) is a controversial decision for menopausal women that has taken on increasing significance as the large number of baby boomers enter this life stage. Studies suggest benefits of HT for prevention of osteoporosis and coronary artery disease, as well as a possible increased risk for cancer, particularly breast cancer. Because of this risk for breast cancer, women with a family history may feel differently about HT. However, differences have not been studied. The purpose of this study was to examine differences in attitudes toward HT of menopausal women with and without a family history of breast cancer. A nonexperimental, cross-sectional design was used. The setting included various sites located in a rural community in northwestern Pennsylvania. A convenience sample of 110 was obtained. A Health Belief Model attitudinal scale was completed by the participants. Although there were no significant differences in attitudes toward HT between the groups, ancillary analysis revealed a significant difference (P = .04) in frequency of reported fear of breast cancer regarding HT in those women with a family history of breast cancer and those without such a history. The findings of this study point to a need for further research on attitudes of women regarding HT and how they may affect postmenopausal healthcare management. PMID- 11858294 TI - Literacy for health information of adult patients and caregivers in a rural emergency department. AB - The nurse practitioner is responsible for providing the patient and family with understandable written information before discharge from the emergency department (ED) following an acute event. Patients are more likely to adhere to their treatment regimen and follow-up if they have a clear understanding of their instructions. Patients cannot correctly interpret information they cannot read. This study describes the ability of two age groups of rural ED patients or caregivers to read health information. The literacy level of 91 younger adults and 104 elderly adults from three rural hospitals was assessed at discharge with the Rapid Estimate of Adult Literacy in Medicine (REALM). While there was no difference in the REALM scores (t = 0.608) of the two age groups, there was a significant difference in the education levels. The REALM scores placed the reading level of the sample at between seventh and eighth grade. The preprinted discharge instruction forms used by the three rural hospitals had a Flesch Kincaid grade level of 9.0, a reading level higher than 62% of the sample. By ensuring discharge instructions are developed at a reading level of sixth grade or below, nurse practitioners can provide teaching materials that encompass the reading levels of the majority of the rural ED population. PMID- 11858295 TI - Care of the patient with chronic pain: part II. AB - Chronic nonmalignant pain frequently results in significant physical, behavioral, psychological, social, and spiritual issues for patients and their families. It is often misunderstood and unsuccessfully managed. Advanced practice nurses who are knowledgeable about chronic pain and the complex biopsychosocial-spiritual needs of this patient population serve an important role in recognizing these patients and intervening appropriately in their care. The purpose of this two part article is to provide that information. Part I [Clinical Excellence for Nurse Practitioners, 3 (4), 192-204] outlined the pathophysiology, assessment, biopsychosocial-spiritual aspects, and pharmacologic treatment of chronic pain. In Part II, a variety of nonpharmacologic and self-management interventions one can use in the primary care setting to treat these difficult health problems are introduced. PMID- 11858296 TI - Quality of life and locus of control of migraineurs. AB - Advanced practice nurses (APNs) are essential primary healthcare providers actively engaged in health promotion, illness prevention, and the management of acute and chronic diseases. The purpose of this study was to measure the quality of life and locus of control (LOC) among three different groups of migraineurs. A retrospective quantitative study was conducted using Ferrans and Powers' (1985) Quality of Life Index and Martin, Holroyd, and Penzien's (1990) Headache Specific Locus of Control questionnaire. The analysis of 79 adult migraineurs indicated that those who received headache education had a higher quality of life and higher external LOC scores than those without the intervention. This study's findings reinforce the notion that disease-specific education is an effective practice intervention. APNs can positively affect the quality of life for migraineurs by providing an educational intervention as part of high-quality, accessible, and cost-effective primary care. PMID- 11858297 TI - A glimpse of nurse practitioners in Wales. AB - Nurse practitioners (NPs) from six nations came together for the 7th International Nurse Practitioner Conference in Cardiff, Wales, in August 1999. The most pressing issues discussed and debated in the international arena continue to be focused on education, certification, and future directions for NPs. PMID- 11858298 TI - To see the world in a grain of sand. PMID- 11858299 TI - Massage as an orthodox medical treatment past and future. AB - Massage has had a long history within orthodox medicine and is not an unknown, untried therapy of dubious origin. Writers on massage usually refer to its long history in ancient cultures but stop with Ling, to whom they attribute the development of modern massage in the early 19th century. Little attention is given to the rich massage literature of the late 19th and early 20th centuries, which reveals massage to have been an orthodox medical therapy practised by doctors and nurses. Analysis of that literature shows massage to have been significantly different in its application to most of today's practice, despite similarity of definition and terminology. Lack of detailed specification of a massage is a limitation of much current research, and this paper suggests a framework, or template, for that specification based on the earlier literature, so that future massages used in research can be replicated and implemented more reliably than at present. Massage as an orthodox therapy almost disappeared after WWII, but has recently reemerged as a complementary therapy which requires a full re-evaluation and audit. The template for its specification, together with the utilization and reconsideration of earlier modes of delivery, offers significant research opportunities for nurses and midwives of today. PMID- 11858300 TI - Moxibustion for breech presentation. AB - Breech presentation at term is considered a possible obstetric complication, and the management before and during labour remains controversial. A technique called 'moxibustion' is used in traditional Chinese medicine to encourage version of the fetus in breech presentation. It has been used in the maternity unit in Plymouth for 11 years. The results would seem to suggest it may have a positive effect and play a part in reducing the number of breech presentations at term and therefore also a reduction in the number of caesarean sections which are so often advocated in breech presentation. This article describes the technique in greater detail and discusses the potential for the future. PMID- 11858301 TI - Aromatherapy for deaf and deafblind people living in residential accommodation. AB - This article looks at ways in which aromatherapy and therapeutic massage have been found to be beneficial for a group of deaf and deafblind adults with special needs, living in residential accommodation. Our basic aim is to promote confidence and communication as well as enhancing a sense of well-being through the judicial use of aromatic plant materials and therapeutic massage. Aromatherapy sessions have become an accepted enjoyable and therapeutic part of the residents' lifestyle. It is our belief that this gentle, non-invasive therapy can benefit deaf and deafblind people, especially as their intact senses can be heightened. This paper explores both professional and caring issues related to the use of aromatherapy in this environment. PMID- 11858302 TI - A concept analysis: the effect of reflexology on homeostasis to establish and maintain lactation. AB - The focus of this paper is to consider whether reflexology can have an effect on homeostasis to establish and maintain lactation. This complementary therapy encompasses the theory of holism to nurture the whole body, mind and spirit (Crane 1997). Parents experience many stressful emotions when their baby is admitted to a neonatal intensive care unit (NICU), which in turn can affect the body, mind and spirit, often contributing to the reduction of lactation for the mother. This paper will utilize a concept analysis approach (Chinn & Jacobs 1987) to provide a focus and sense of direction to the discussion and exploration of the topic. PMID- 11858303 TI - Working with Alice: a reflection. AB - Reflective practice has become established within educational programmes and in clinical supervision. Yet what does it mean to be a reflective practitioner? This paper seeks to give some meaning to this concept by drawing on a story I wrote in my reflective journal one evening. My reflection on this event draws out key issues of practice and reflection that enable me to gain insight and apply to future practice within a reflexive learning spiral. PMID- 11858304 TI - T'ai chi ch'uan. AB - The Chinese practice of t'ai chi seems to be receiving increased interest in the West. This article gives a brief overview of t'ai chi, including its origins, development, principles and potential health benefits. The function of the essential elements of t'ai chi, namely the Form and chi kung are described and their potential benefits for patients and nurses are discussed. Exponents of t'ai chi believe that it has health benefits on physical, psychological and spiritual levels, thus promoting a feeling of well-being. In addition, regular practitioners are empowered to be in greater control of themselves, their health, and situations in which they find themselves. PMID- 11858305 TI - Shamanism and complementary therapy. AB - Shamanism is an ancient tradition which may offer profound insights into the healing process and to our whole understanding of health. It has an extensive historical and geographical distribution, and may contain elements essential to our understanding of humanity. This paper outlines the origin and nature of shamanic practice, and considers its implications for a number of current healing issues. Several areas of potential relevance to complementary practitioners are explored. These include the shamanic concepts of illness, change and growth; illness and healing as rites of passage; death and dying; and the use of imagery in healing. This paper suggests that complementary practitioners may find some shamanic principles highly congruent with their own practice. PMID- 11858306 TI - Cleaning up. The development of the Alfred/Medseed Wound Imaging System. AB - This paper describes the rationale, development, functions and applications of a new digital wound imaging system developed at The Alfred Hospital in Melbourne. The system known as The Alfred/Medseed Wound Imaging System (AMWIS) comprises a software package that quantitatively analyses a digital wound photograph and tracks the healing of the wound as a function of the wound care treatment used. The system is designed to enable the wound images and assessment data to be securely transmitted via the internet for review or consultation to any site equipped with the wound imaging system. The AMWIS is capable of being operated at a single site such as a hospital, at remote locations or as a mobile system. We believe that this is the first system of this kind developed in Australia that incorporates sophisticated analytical functions yet retains the flexibility to be used in inpatient, ambulatory or remote settings. PMID- 11858307 TI - Academic maturity: qualifications to teach the nurse professionals of the future. AB - In a recent survey of Queensland academic managers and graduates of pre registration and nursing postgraduate programs, it was found that within levels of management there exists a disparity related to the minimal qualifications of educators of nurses. The preferred minimal level of academic preparation of heads of departments of nursing was generally ess than that proposed by recent consumers of tertiary education. Recent postgraduate and graduate students displayed their academic maturity through their preferences for highly qualified and academically qualified educators at all levels. The variety of educators involved in tertiary education included academics teaching within universities; clinical laboratory teachers teaching the skills of nursing practice; clinical faci itators who are responsible for groups of students in the clinical area; and clinical mentors, the clinicians who support and nurture students on a one-to-one basis. Ten managers of nursing programs (N=20), fourteen postgraduates (N=61) and 90 graduates (n=302) of Queensland tertiary nursing programs responded to the survey. While the final samples were small, the outcomes suggest increasing graduate expectations of thei educators and the need for a re-evaluation of measures of quality in academic teaching. PMID- 11858308 TI - The language used when reporting interfemale violence among nurses in the workplace. AB - Violence appears to be ignored among nurses, in part, because of the metaphors and euphemisms used to identify this significant phenomenon. Nurses rarely use the term 'violence' and may use metaphors and euphemisms or generally ignore the terms usually associated with violence, such as victim, perpetrator, horizontal violence, bullying and mobbing when discussing violent behaviour among, and with, their female colleagues. The reluctance to use unequivocal language when discussing workplace violence contributes to the issue remaining poorly recognised and poorly addressed; and increases the negative impact of workplace violence on the health and wellbeing of the individual. Moreover, the language that is used to describe violence among female nurses often carries with it different connotations from those that may be common in general society. This paper draws upon data from the author's doctoral research study to report the language used to describe interfemale violence in the workplace; and the consequences of using that language. PMID- 11858309 TI - The experiences of the registered nurse caring for the person dying of cancer in a nursing home. AB - This paper reports on research that set out to identify and describe the experiences of the registered nurse caring for the resident dying of cancer in a nursing home. The research method used was a qualitative single case study involving five registered nurses in one nursing home. Thematic analysis of data from unstructured interviews and reflective journals was used to explicate the nurses' experiences. Four major themes were extracted from the data. These themes were the exclusivity of the relationships, difficulties in the management of pain, the expectations of the registered nurse and the impact of caring. The nurses' experiences were loving, fascinating, compelling and rich. The overarching experience of the nurses came from the relationship they formed with the residents. A crucial finding in this study was that these registered nurses valued the elderly resident and developed meaningful relationships with them. All aspects of the relationships were unique and profound. However, the emotional involvement with the resident, multiple death losses, confrontation of personal losses, limited workplace support such as counselling, potentially places these registered nurses at emotional risk from burnout and complicated bereavement. The implications of these findings for nursing practice, education and research are discussed. PMID- 11858310 TI - The contribution of nurses' perceptions and actions in defining scope and stabilising professional boundaries of nursing practice. AB - This paper reports on the findings of a naturalistic inquiry study that explored the scope and boundaries of nursing practice. Findings from interview and observation data suggest that nurses negotiate and adjust professional boundaries on an individual, case-by-case basis, thereby managing the scope of their practice as they see it in that circumstance. The strategies they used are presented in four major categories: 1) maintaining a comfort zone, 2) expanding into safe territory, 3) moving into the grey zone and 4) stepping over the line. Findings show that nurses' efforts to maintain the comfort zone serve to perpetuate the status quo and may threaten holistic care. Expanding nursing actions to include functional roles such as coordinating care, sharing information, advocating (for patients), collaborating and innovating offers the profession critical building blocks for defining the scope of nursing practice. Clarifying the grey zone (or overlapping territory) is an essential task for the profession in determining the boundaries of nursing practice. The data revealed that, partly due to the ambiguity of the grey zone, nurses may step over the line into medical decision-making and outside the legal sanctions for the professional nursing role. The implications of this study highlight the need for nursing to define its scope of practice and in so doing stabilise professional boundaries. PMID- 11858311 TI - Urinary continence promotion: a role for all nurses. PMID- 11858312 TI - A clinical teaching project. Evaluation of the mentor-arranged clinical practice by RN mentors. AB - This paper explores the relationship between nursing students and mentors and its influence on nursing students' development of professional knowledge during their clinical practice. The paper is based on an evaluation of a new initiative supplementing other clinical experiences for students of nursing, Mentor-Arranged Clinical Practice (MACP), by registered nurse mentors. Sixty-five (95.6%) mentors returned their questionnaires in a self-addressed envelope. Results of the study indicated that there were benefits and rewards for both students and mentors. The mentors felt that students gained more practicum time, confidence and independence in their practice, time management skills, rapport between staff and student, and better integration into the hospital environment. Mentors indicated that the most compelling reasons for becoming mentors were the opportunity to assist mentees to integrate into the nursing unit, to teach, share knowledge, promoting and improving the professional image of nurses, and gain job satisfaction from mentoring. The findings in this study suggest that there is a need for development of mentor relationships in the clinical setting of undergraduate nursing courses. PMID- 11858313 TI - Domestic violence around the world: when will it end? PMID- 11858314 TI - Hosting a wellness day: promoting health in the old-old. AB - The purpose of health promotion and disease prevention is to increase the number of years of life and ensure better quality of remaining life. Those in the oldest age group (> or = 90 years) are generally less likely to monitor their cholesterol intake, exercise, have their stools checked for occult blood, or undergo a mammogram, Papanicolaou test, or prostate or skin evaluation as recommended. The most common reasons given for not engaging in these activities are advanced age, absence of direction by primary health care providers, and lack of interest in following up abnormal findings. To promote primary and secondary disease prevention and health promotion, a Wellness Day was hosted in a continuing-care retirement community by the in-house geriatric nurse practitioner and a group of nurse practitioner students. The program offered cancer screening, cardiovascular health assessments, and exercise prescreening and education, as well as screening for diabetes and risk for osteoporosis. An individualized approach to health promotion was used, reviewing with each resident the pros and cons of participating in each of these behaviors. PMID- 11858315 TI - Cultural perspectives on contraception: a literature review. AB - The incorporation of transcultural nursing concepts has become an integral component of holistic nursing practice. However, to deliver culturally appropriate nursing care, knowledge of cross-cultural health beliefs and practices is necessary. This article reviews cultural perspectives on contraception based on a literature review of qualitative and quantitative health care research studies related to the topics of culture and contraception. The purpose of this literature review is to provide a knowledge foundation for health care providers who provide contraceptive counseling. Implications for practice, as well as recommendations for future research, are developed. PMID- 11858316 TI - Polycystic ovary syndrome: a case presentation. AB - Polycystic ovary syndrome (PCOS) is a disorder of excessive adrenal hormone production that results in characteristic features. It is associated with hyperinsulinemia and, if left untreated, carries a risk for developing such complications as diabetes and heart disease. The clinical features are described here by means of a case presentation. Using the Ferriman-Gallwey score, the degree of hirsutism is identified. Differential diagnoses and recommended treatments are discussed. The role of the advanced practice nurse in the diagnosis and care of patients with PCOS is included. PMID- 11858317 TI - Pain in the elderly: polymyalgia rheumatica. AB - Diagnosis of rheumatologic disorders in the elderly is often complicated by the primary care clinician's inability to differentiate among similar manifestations of rheumatologic disorders, the presence of comorbid conditions, and symptoms attributed simply to aging. A major consequence of the aches and pains associated with rheumatologic disorders, including polymyalgia rheumatica (PMR), is the impedance of activities of daily living, potentially leading to a loss of independence. PMR is common in the elderly. Often coexisting with PMR, temporal arteritis can lead to complications, including blindness, stroke, or cardiac sequelae. Timely detection and appropriate treatment of PMR in the elderly may improve quality of life, as well as deter irreversible problems. Patient education also has an important role. PMID- 11858318 TI - Insights into genetic testing for colon cancer: the nurse practitioner role. AB - As new genetic discoveries continue to gain public awareness, patients will increasingly call on their nurse practitioners (NPs) to discuss their inherited susceptibility to disease. Genetic testing for colon cancer can presently identify gene mutations for 2 inherited forms of this disease, familial adenomatous polyposis and hereditary nonpolyposis colorectal cancer, accounting for approximately 6% of the cases. By identifying patients at high risk for colon cancer, NPs can discuss the benefits of early detection through screening procedures while helping patients gain insight into the meaning and impact genetic testing can have on their lives. This article discusses the basic genetics involved and screening recommendations for those with a hereditary disposition to colon cancer. Benefits, risks, and limitations are also considered, along with the importance of the NP in educating and supporting individuals in their decision making about genetic testing for colon cancer. PMID- 11858319 TI - Ovarian cancer: a case study. AB - Ovarian cancer is the leading cause of death from gynecologic malignancies in the United States, accounting for approximately 15,000 deaths each year. Patients commonly present to their primary care providers with vague abdominal symptoms. Health care providers need to become more aware of the importance of pursuing seemingly vague symptoms. A discerning knowledge of the risk factors and symptoms of this disease enables the clinician to institute appropriate assessment strategies, intervention, and referral, resulting in reduced mortality rates. This case report of a white woman aged in her 60s diagnosed with ovarian cancer stresses the need for early screening and detection, disease education, and prompt referral and management to displace the fear associated with the diagnosis and improve survival. PMID- 11858320 TI - Pertussis in the adolescent and adult: a primary care concern. AB - Pertussis, a highly communicable, infectious respiratory disease, is considered a disease of childhood. The introduction of the standardized whole-cell pertussis vaccine in 1949 nearly eradicated pertussis. In 1976 in the United States, the lowest number of pertussis cases ever was reported. However, in recent years, there has been a resurgence in the number of pertussis cases among all age groups. This increase is especially noted among adolescents and adults. In 1996, the U.S. Centers for Disease Control and Prevention reported 7,796 cases of pertussis, with almost half among individuals aged 10 years or older. Although pertussis in adolescents and adults is generally more mild than in children, it is a major source of disease transmission to younger children, who may have more serious complications. Consequently, early recognition and treatment of pertussis in adolescents and adults is imperative in preventing potentially life threatening illness in infants and young children. This article examines pertussis in the adolescent and adult population, describing the epidemiologic characteristics of the disease, its clinical manifestations, diagnosis, treatment, and the use of vaccinations. PMID- 11858321 TI - NPACE nurse practitioner practice characteristics, salary, and benefits survey: 1999. Nurse Practitioner Associates for Continuing Education. AB - The purpose of this article is to present the results of a practice characteristics, salary, and benefits survey of 1,557 nurse practitioners from the United States who attended national nurse practitioner conferences in Las Vegas, Nevada, Orlando, Florida, Chicago, Illinois, and Boston, Massachusetts, in 1999. Specific data are presented on the demographics of the population, practice characteristics and responsibilities, benefits for full- and part-time employees, and salary by region, years of practice, type of certification, and location of the practice. The salary data were compared with the 1995-1996 and 1996-1997 NPACE practice characteristics, salary, and benefits surveys (Pulcini & Fitzgerald, 1997; Pulcini, Vampola, & Fitzgerald, 1998). PMID- 11858322 TI - The complex challenge of HIV/AIDS care: a global perspective: an update from XIIIth International AIDS Conference. AB - This article provides an overview of important findings and discussion from the XIIIth International AIDS Conference held in Durban, South Africa, July 2000. It addresses global epidemiology, the sociopolitical impact of HIV disease, newer strategies to combat disease including medications and microbicides, advances laying the groundwork for vaccine development, and concerns about the economics of care. It highlights the need to recognize the complexity of the threat to the well-being of the entire global village posed by this epidemic. PMID- 11858323 TI - Wade, G. H. (1999). Professional nurse autonomy: concept analysis and application to nursing education. Journal of Advanced Nursing, 30 (2), 310-318. PMID- 11858324 TI - Bowen, K. (2000). Child abuse and domestic violence in families of children seen for suspected sexual abuse. Clinical Pediatrics, 29 (1), 33-40. PMID- 11858325 TI - Implementation of PET scan as a new service. AB - Positron emission tomography (PET) is a diagnostic test using injected radiopharmaceuticals (drugs or compounds tagged with radioactive isotopes) to obtain images of metabolic/physiologic processes. PET provides physiologic data rather than just anatomic data such as that provided by CT scan or MRI. Metabolic changes precede anatomic changes, hence the higher accuracy of PET. Scar tissue, necrosis and tumor mass may appear identical on CT and MRI, but markedly different on PET. PET may decrease unnecessary surgeries and improve treatment through greater specificity about the disease and may provide a better evaluation of recurrent disease. PMID- 11858326 TI - The revolution in radiation therapy. PMID- 11858327 TI - Coding update. I'm sure it's all a terrible mistake... PMID- 11858328 TI - The effects of tibial malrotation on the biomechanics of the tibiotalar joint. AB - The effects of tibial malrotation on the biomechanics of the tibiotalar joint were studied using a cadaveric model loaded in an Instron 8521 materials testing device and a TEKScan I-Scan thin-film resistive ink pressure measuring system. Testing of 23 legs was performed using rotational conditions of 10 and 20 degrees internal and external rotation as well as neutral rotation. All rotational conditions were found to decrease joint contact area. Peak pressures were significantly greater with 20 degrees internal rotation as well as 20 degrees external rotation. Total load across the joint was significantly lower for both 10 and 20 degrees of external rotation. In conclusion, rotational deformity across the tibiotalar joint results in significant alteration of overall joint biomechanics and should be minimized whenever possible. PMID- 11858330 TI - The Orthopaedic Research Institute-Ankle Strength Testing System: inter-rater and intra-rater reliability testing. AB - Achilles tendinosis is a degenerative overuse tendinopathy involving the primary ankle plantarflexors, namely the soleus and gastrocnemius muscles forming the tendo Achilles. The Orthopaedic Research Institute-Ankle Strength Testing System (ORI-ASTS) was designed to record objective measurements of force generated with a resisted ankle plantarflexion test. Testing normal subjects (n=6) was used to establish the reliability of the ORI-ASTS for measuring ankle plantarflexion force. Testing patients with Achilles tendonitis (n=5) over time and comparing Achilles tendon analogue pain scores to ankle plantarflexion force measurements was used to assess the validity of the ORI-ASTS for monitoring progression of Achilles tendinopathy. Inter-rater reliability of the ORI-ASTS with normal subjects was excellent, with single measure intraclass correlation coefficients (ICC) for right mean peak force of 0.92, left mean peak force of 0.96, right mean total force of 0.89, left mean total force of 0.91. The ORI-ASTS also had excellent intra-rater reliability for normal subjects with the following single measure ICC scores: right mean force 0.96, left mean force 0.92, right mean total force 0.97, left mean total force 0.92. The relative technical errors of measurement were calculated from these results, and ranged from 1.0% to 2.7%. Testing patients with Achilles tendinosis (n=5), and comparing analogue activity pain scores to ORI-ASTS testing demonstrated a strong negative relationship between the two parameters (Spearman Rho -0.87 to -1.0, Kendall tau b -0.82 to 1.0). The relationship was statistically significant at the p=0.01 level for two of the five patients. The ORI-ASTS shows excellent reliability for testing ankle plantarflexor force, and appears valid for objectively assessing and monitoring patients with Achilles tendinosis. PMID- 11858329 TI - The effects of rotation on radiographic evaluation of the tibiofibular syndesmosis. AB - Radiographs of 12 normal cadaveric lower extremities were prepared with each extremity in seven increments of axial rotation, ranging from 5 degrees of external rotation to 25 degrees of internal rotation. The tibiofibular clear space, the tibiofibular overlap, the width of the tibia and fibula, and the medial clear space were measured on each film. The width of the tibiofibular clear space (syndesmosis A) averaged 3.9+/-0.9 mm (range, 2 to 5.5 mm), but did not change significantly with rotation. Its size was independent of the size of tibia and fibula. All other measurements changed dramatically with rotation. In our specimens, a true mortise view of the ankle joint was obtained by internally rotating the extremity an average of 13.6+/-0.7 degrees (range, 12.0 degrees to 17.0 degrees). Based on our results the width of the tibiofibular clear space on the anterior-posterior view is the most reliable parameter for detecting widening of the syndesmosis on plain radiographs. However, due to its variability among different individuals, comparison views of the contralateral extremity are warranted for confirmation of clinical suspicion of syndesmosis disruption. PMID- 11858331 TI - The relationship of the anterior drawer sign to the shape of the tibial plafond in chronic lateral instability of the ankle. AB - To determine what factors correlate with values for tested anterior drawer (AD) sign, we investigated stress radiographs in 71 patients with severe chronic lateral instability of the ankle as well as 320 normal controls. We found no correlation between AD and talar tilt (TT), but measurement of the anterior tip ratio (ATR) (ATR=tanA x 100) demonstrated a link of ATR with AD (r=0.74, p<0.001). The posterior tip ratio (PTR) (PTR=tanP x 100) was not correlated with AD (r=0.34, p=0.16). The ankle mortise angle (AMA) (the anterior opening of the lateral ankle mortise) was also not correlated with AD (r=0.27, p=0.23). Investigation of 320 normal ankles revealed a mean ATR of 9.1%+/-1.4%. The ATR in patients was 7.6%+/-1.7%, significantly lower than in the control patients. The values in female patients with chronic lateral instabilities were significantly less than the values in the females in the control group. But values for male patients were not different from the control group. Thus the anterior tip of the lateral tibial plafond, as it affects the ATR, may influence instability in anterior drawer function, especially in female patients with chronic lateral ligamentous instability of the ankle. PMID- 11858332 TI - Proprioception of the ankle: a comparison between female teenaged gymnasts and controls. AB - Proprioceptive mechanisms appear to play a role in stabilizing the joints and may serve as a means for interplay between static stabilizers and dynamic muscular restraints. The purpose of our study was to investigate whether or not gymnastic training has any effect on the balance and on proprioception in an ankle, as seen in gymnasts and in nongymnasts. We evaluated the proprioceptive ability of the ankle using four different tests (a one-leg-standing test, a single-limb-hopping test, an active angle-reproduction test, and a passive angle-reproduction test). Proprioception of the ankle was measured in 40 subjects who were assigned to two experimental groups. Group 1 (n: 20) were healthy control subjects, and group 2 (n: 20) were teenaged female gymnasts. The sense of position of a joint was actively measured using a Cybex NORMTM isokinetic dynamometer and measured passively with a proprioception-testing device. A Mann-Whitney U test was used to compare mean values of the gymnasts to the controls. Results revealed statistically significant differences (p<0.05) between two groups. We found no statistically significant differences between the dominant and nondominant ankle in volunteers or in gymnasts, in all tests. The results of this study suggest that gymnastic training has a positive influence on sense of position of the ankle joint and on balance, in addition to increasing muscle tone. PMID- 11858333 TI - Single stage correction with external fixation of the ulcerated foot in individuals with Charcot neuroarthropathy. AB - The ulcerated foot in individuals with Charcot neuroarthropathy presents a complex problem when correction of the deformity is necessary but the presence of infection precludes the use of internal fixation. We reviewed 11 patients with midfoot Charcot neuroarthropathy, collapse, and ulceration who were at risk for amputation. These patients underwent operative debridement, corrective osteotomy, external skeletal fixation and culture-directed antibiotic therapy as a limb salvage procedure. Patients were transitioned from the external fixator (average 57 days) to total contact casting (average 131 days) and all subsequently progressed to therapeutic footwear in 12 to 49 months of follow-up (average 24 months), except one patient whose medical decline resulted in bedrest. We believe that when performed in properly selected patients, this procedure presents an alternative to amputation and, via corrective osteotomy, results in a shoe-able, functional foot that is potentially less prone to ulceration. PMID- 11858334 TI - Management of the neuromuscular foot deformities with the Ilizarov method. AB - Nine feet of eight patients with neurologic foot deformities caused from poliomyelitis, Charcot-Marie-Tooth disease, and traumatic nerve injury were treated with V-osteotomy and the Ilizarov method. A painless and plantigrade foot was obtained in all but one patient. A residual deformity occurred in this case because of the soft-tissue problems during the correction period. No major complication had been encountered by the latest follow-up at a mean of 42 months. The results of the current study indicate that the V-osteotomy and the Ilizarov method, while technically difficult, is an effective method for correcting neurologic foot deformities and obtaining a stable, plantigrade, painless foot, especially when complicated with other lower leg problems (i.e., leg length discrepancy, joint contractures). PMID- 11858335 TI - Heel pad thickness--a contributing factor associated with plantar heel pain in young adults. AB - Thirty-three runners were clinically diagnosed with plantar heel pain (PHP) and 107 runners were designated into a non-PHP group (NPHP). Cases were matched with a further control group of 64 subjects with regard to body mass index and age. A weight-bearing platform measured heel pad thickness by real-time ultrasound scanning. The results demonstrated a significant difference between the PHP and both the NPHP and control groups (P <0.05). The results also demonstrated that body mass index had a significant effect (P <0.05) on the heel pad thickness of all subjects, regardless of the heel pad group. The results from the current study, with some precaution, conclude that body mass index as such is not related to PHP in the athletic population. In the current study the observed differences of body mass index on heel pad thickness between the PHP and NPHP groups, and the similarity between the control group and NPHP subjects suggests other risk factors may play an important role in heel pad thickening and associated development of PHP. PMID- 11858336 TI - A comparison of two analytical techniques for detecting differences in regional vertical impulses due to plantar fasciitis. AB - The vertical impulse distribution beneath the foot has been shown to be a useful objective method for evaluating gait and the efficacy of treatment programs. However, recent studies employing similar methods, but different analytical techniques, have reached divergent conclusions regarding the effect of plantar fasciitis on hindfoot, midfoot and forefoot impulses. The aim of the current study was to determine whether the impulse distribution beneath the hindfoot, midfoot and forefoot in subjects with, and without, plantar fasciitis was dependent on the analytic technique employed. A pressure platform was used to collect impulse estimates from 16 subjects with plantar fasciitis, and 16 control subjects, while walking at their preferred speed. The findings indicate that an impulse calculation incorporating the position of the center of pressure is more effective in detecting alterations in gait than the conventional method of estimating the impulse distribution beneath the foot. This study also demonstrates that subjects with plantar fasciitis possess modified gait patterns that are primarily manifest by a reduced hindfoot and an increased midfoot impulse, as defined by the pathway of the center of pressure. PMID- 11858337 TI - An unusual case of bilaterally symmetrical neuropathic osteoarthropathy of the midfoot as a result of lyme disease-induced peripheral neuropathy: a case report. AB - The first known report of a patient with Lyme Disease-induced peripheral neuropathy resulting in bilaterally symmetrical "Charcot" changes of the midfoot is reported. An extensive review of the literature failed to produce any known cases of neuropathic osteoarthropathy that would be linked directly or indirectly with Lyme Disease-induced peripheral neuropathy. The relationship between Lyme Disease, the secondary neurologic manifestations caused by the infection, and neuropathic osteoarthropathy are discussed. PMID- 11858338 TI - Microscopic metallic wear and tissue response in failed titanium hallux metatarsophalangeal implants: two cases. AB - The membranes present at the implant-bone interface were retrieved from two patients with titanium single stem hallux implants that had failed. Both patients had pain and valgus deformity of the hallux, and radiographs showed a radiolucent shadow around the implant stem, with thinning of the dorsal cortex of the proximal phalanx in one patient. After removal of the implants, arthrodesis of the first metatarsophalangeal (MP) joint was performed. Histologic analysis of the membrane tissue at the implant-bone interface showed a synovial-like appearance. There was a fibrous tissue stroma adjacent to the bone surface, with multiple regions of scalloping covered by mononuclear cells. Fine metallic debris was seen throughout the fibrous tissue. Multinucleated foreign body giant cells were sparsely observed associated with fine particulate metallic wear debris similar to observations from failed total hip arthroplasties. The histologic appearance is evidence that foreign-body granulomatous infiltration associated with metallic wear debris may be a causative factor of peri-implant osteolysis leading to aseptic loosening and failure of titanium single stem hallux implants. PMID- 11858339 TI - Pericuboid fracture-dislocation with cuboid subluxation. AB - The case of a 44-year-old male with a traumatic pericuboid fracture-dislocation consisting of disruption of the midtarsal, tarsometatarsal and pericuboid articulations is presented. The cuboid was partially dislocated without cuboid fracture. Treatment consisted of open reduction with stable internal fixation followed by early range of motion and protected weight-bearing for 12 weeks. Functional outcome results were excellent at final follow-up. PMID- 11858340 TI - Report on the A.O.F.A.S. Outreach and Educational Fund fact-finding mission to Vietnam. American Orthopaedic Foot & Ankle Society. PMID- 11858341 TI - Wild man and a dozen red roses. PMID- 11858342 TI - Arthrodesis of the first metatarsophalangeal joint: a biomechanical study comparing memory compression staples, cannulated screws, and a dorsal plate. AB - Arthrodesis of the first metatarsophalangeal joint of 21 matched pairs of cadaver toes was performed in order to compare the strength of three methods of internal fixation: 1. two crossed cannulated screws, 2. a dorsal plate with an oblique 0.062 K-wire, and 3. two compression staples with an oblique 0.062 K-wire. Biomechanical testing with plantar force was carried out, and gapping across the fusion site was measured. Stiffness, load to 1-mm displacement, and force to failure was determined for each specimen. Both the plate and screw constructs were statistically stronger in force to failure and initial stiffness than the compression construct. Compression staples have an advantage in their ease of insertion and theoretical continuous compressive force across an arthrodesis site, but should be supplemented with a cast or other external immobilization until union is achieved. PMID- 11858343 TI - An irritant. PMID- 11858344 TI - Decreasing effects of iron toxicosis on selenium and glutathione peroxidase activity. AB - Heart failure due to chronic iron overload is a leading cause of cardiovascular mortality in the second and third decades of life worldwide, but its mechanism is not known. Deficiencies of selenium have been shown to result in damage to the myocardium and to the development of various cardiomyopathies. In the current investigation, the dose-dependent effects of chronic iron toxicosis on heart tissue concentrations of selenium and the protective antioxidant enzyme glutathione peroxidase (GPx) were investigated in a murine model of iron-overload cardiomyopathy (n = 20). Significant dose-dependent decreases in heart tissue selenium concentrations (r = -0.95, p < 0.001) and selenium-dependent GPx activity (r = -0.93, p < 0.001) were observed in chronically iron-loaded mice in comparison with placebo controls. These results suggest that dietary supplementation with selenium may be beneficial in the clinical management of disorders of iron metabolism. PMID- 11858345 TI - Monitoring hydration status in elderly veterans. AB - Dehydration is a common water and electrolyte disorder in long-term care residents. Practical methods to detect and monitor dehydration are needed. This study determined whether urine color measured by a urine color chart reflects hydration status. The study employed a repeated measures design with two observations during a 10-hour period. Urine color was compared to the criterion standard of urine specific gravity and osmolality. The sample included 89 participants from two Veterans' Affairs facilities. Urine color was graded on an eight-level color chart. Urine specific gravity and osmolality, serum sodium and osmolality, hematocrit, blood urea nitrogen (BUN), and creatinine were measured using standard laboratory procedures. Significant positive associations existed between urine color and both urine specific gravity and urine osmolality and between urine osmolality and serum sodium and the blood urea nitrogen to creatinine ratio. Although further testing is needed, the color chart has potential as a low-cost technology to monitor dehydration. PMID- 11858346 TI - Advance directives and community-dwelling older adults. AB - Advance directives (ADs) are documents that allow competent individuals to set forth their medical treatment wishes and/or to name a proxy in the event that they lose the capacity to communicate these decisions in the future. Despite the benefits of and support for such documents, very few people have completed an AD. This posttest-only experimental study examined whether an individualized intervention given to half of the older adults who attended an educational session increased the discussion and/or completion of ADs. Of the 74 participants, 25.7% (n = 19) completed an AD. There were no significant differences between control and intervention groups on the discussion and/or completion of ADs. Multivariate analysis indicated that perceived barriers were significantly associated with the discussion and completion of ADs. Content analysis revealed that major barriers to discussion and completion include procrastination and a reluctance to think about deteriorating health status and/or death. PMID- 11858347 TI - Mexican-origin mothers' experiences using children's health care services. AB - A focused ethnographic study in an urban Latino community in the western United States describes Mexican-origin mothers 'experiences obtaining and using health services for their children. Repeated interviews with mothers, participant observation, and children's medical records composed the data sources. Qualitative findings suggest access to health care begins in the household, where women negotiate a working diagnosis for the children's illness with family members and coalesce support for health care seeking. Immigrant mothers described more barriers to children's health care than more acculturated mothers. Quantitative analyses of medical records supported this finding, with children of the least acculturated mothers demonstrating fewer well-child visits, increased emergent visits, and lower levels of immunization completeness. The results suggest health care providers can better meet the needs of Latino families with children by offering better explanations about children's diagnoses and treatment plans and demonstrating personalismo, or a friendly, kind, and social approach to care. PMID- 11858348 TI - Spouses' experiences in heart transplantation. AB - In this phenomenological study, in-depth interviews were used to obtain a description of spouses' experiences in heart transplantation. Thematic analysis of the transcripts revealed four major, interrelated themes: death-life, vigilance, change, and gift. The experience was contextualized by the existential grounds of time and other people. Findings suggested that the changes inherent in the transplant experience have not been fully described in previous studies. The theme of death-life was dominant and pervasive in all interviews. As the threat of their husbands' deaths became less prominent, wives reported difficulty letting go of their vigilance. Although the most outstanding gift was that of the donor organ, spouses also gratefully remembered the donor and donor family. Time was not perceived as a boundary separating periods of the transplant experience; all aspects of time seemed to be woven seamlessly from the past through the present and into the future. Spouses were adamant that health care personnel should recognize them as coparticipants in the critical life event of cardiac transplantation. PMID- 11858349 TI - Caregiver singing and background music in dementia care. AB - Caregiver singing and background music were incorporated into the interaction between caregiver and patient, the aim being to illuminate the meaning of verbal communication between persons with severe dementia and their caregivers. In the absence of music, patients communicated with cognitive and behavioral symptoms associated with dementia. In these situations, caregivers devoted their verbal communication to narrating and explaining their caring activities to the patient. The patient and caregiver, however, had difficulties understanding one another. In the presence of background music, caregivers decreased their verbal instructing and narrating while the patient communicated with an increased understanding of the situation, both verbally and behaviorally. During caregiver singing, a paradoxical effect was observed such that despite at evident reduction in the amount of verbal narration and description by the caregiver; the patient implicitly understood what was happening. PMID- 11858350 TI - Potential impact of pharmacogenomics on the future of drug development and practice of rheumatology. A look from the trenches. PMID- 11858351 TI - Discovery of TNF-alpha as a therapeutic target in rheumatoid arthritis: preclinical and clinical studies. AB - The development of effective new treatments is greatly facilitated by the understanding of the mechanisms of disease. In rheumatoid arthritis, there has been progress in understanding its immunology, the HLA class II predisposition including the 'shared epitope' and more recently in understanding the importance of proinflammatory cytokines. Here we review our work in defining TNFalpha as a therapeutic target in rheumatoid arthritis, from an understanding of molecular pathogenesis in vitro, to formal proof in the clinic in vivo. There is now extensive clinical use of anti-TNFalpha biologicals for severe rheumatoid arthritis in the US and Europe. PMID- 11858352 TI - Bisphosphonates in bone diseases other than osteoporosis. AB - The range of bone diseases in which bisphosphonates are used has extended far beyond osteoporosis during the last few years. Bisphosphonate therapy is now so well validated as to be the reference standard in Paget's disease and in the prevention of bone complications related to malignant osteolysis. Promising preliminary findings warrant the use of bisphosphonates in conditions that are either rare (fibrous dysplasia) or severe (pediatric osteogenesis imperfecta). The third category of indications encompasses many conditions in which the limited available data do not warrant widespread use: examples include reflex sympathetic dystrophy syndrome, acute back pain after a vertebral crush fracture, and chronic inflammatory joint disease not treated by glucocorticoids. PMID- 11858353 TI - Parathyroid gland radionuclide scanning--methods and indications. AB - The usefulness of preoperative radionuclide scanning of the parathyroid glands in patients with primary or secondary hyperparathyroidism was long controversial because available techniques were of limited diagnostic efficacy. Technetium-99m labeled sestamibi (99Tc-sestamibi) is a new radiopharmaceutical agent easily detected by gamma cameras. The first parathyroid imaging studies done with 99Tc sestamibi about 10 years ago used a double-phase technique to separate thyroid and parathyroid tissue. Although promising, this method was less than ideal, particularly in multiple gland primary hyperparathyroidism and in secondary hyperparathyroidism. For several years, we have been using subtraction between two images acquired simultaneously, one with 99Tc-sestamibi, which binds to thyroid and parathyroid tissue, and the other with 123-iodine, which binds only to thyroid tissue. The remarkable efficacy of this technique in both primary and secondary hyperparathyroidism invites a reappraisal of the place of radionuclide imaging as a preoperative localization procedure done to reduce the need for repeat surgery. The usefulness of this technique in selecting candidates for unilateral surgery among patients with primary hyperparathyroidism is discussed. PMID- 11858354 TI - Second-line drugs used in recent-onset rheumatoid arthritis in Brittany (France). AB - OBJECTIVE: The management of recent-onset rheumatoid arthritis (RA) is not well standardized. We conducted a survey of drugs prescribed to RA patients in Brittany at presentation and during the first 1 to 3 years of follow-up. METHODS: A cohort of 270 patients with recent-onset inflammatory joint disease was recruited between 1995 and 1997. The evaluation at presentation included a medical history, a thorough physical examination, and a standard battery of investigations. Follow-up at 6-month intervals was offered. At the last visit, between June and December 1999, a panel of five rheumatologists established that 98 patients had RA. RESULTS: At presentation, hydroxychloroquine and injectable gold were the most widely used second-line drugs, and only two patients were offered a combination of second-line drugs. At the last visit, the most commonly used drugs were methotrexate, injectable gold, and hydroxychloroquine (23, 23, and 21 patients, respectively); only three patients were on more than one second line drug and 38 (38/98, 39%) patients were on glucocorticoid therapy. CONCLUSION: Rheumatologists in Brittany prefer monotherapy with hydroxychloroquine or injectable gold as the initial treatment. Later, they rely mainly on methotrexate, injectable gold, and hydroxychloroquine, often in combination with glucocorticoid therapy. PMID- 11858355 TI - Is there a place for non-selective NSAIDs in the treatment of arthritis? PMID- 11858356 TI - Anti-idiotype antibodies abrogate the tissue deposition of anti-RNP human autoantibodies injected into neonatal BALB/c mice. AB - OBJECTIVE: The goals of the current work are: 1) to examine the epidermal deposition of anti-RNP IgG human autoantibodies in neonatal BALB/c mice; 2) to look for immunoregulatory effects of anti-idiotypes allowing one to inhibit the epidermal deposition of anti-RNP antibodies; and 3) to elicit antinuclear antibodies in adult BALB/c mice by internal images of anti-idiotypes. MATERIALS AND METHODS: Anti-idiotype antibodies were produced with human anti-RNP IgG obtained by ion exchange chromatography; F(ab')2 fragments were recovered from pepsin digestion and were purified using Sephacryl S-300. F(ab')2 fragments were then used to immunize New Zealand rabbits. RESULTS: The anti-RNP IgG recognized the 70 kDa protein and the A (31 kDa) and C (19 kDa) proteins, while the anti idiotype antibody specifically recognized the light or heavy chain of the anti RNP (Fab')2 fragments. Additionally, anti-idiotypes recognized the anti-RNP IgG from some sera, but not the IgG from other specificities or from normal IgG. When anti-RNP IgG was injected intraperitoneally into BALB/c mice it induced immune complex deposition in the epidermis and at the dermal-epidermal junction. Previous injection of anti-idiotype antibodies abrogated the anti-RNP IgG deposits. Vaccination with anti-idiotypes elicit antinuclear antibodies in adult BALB/c mice. CONCLUSIONS: Anti-idiotype antibodies abrogate in vitro the antinuclear antibody deposition in neonatal BALB/c mice. Anti-idiotype antibodies elicit antinuclear antibodies in adult BALB/c mice. PMID- 11858357 TI - Fibromyalgia in Italian patients with primary Sjogren's syndrome. AB - OBJECTIVE: To assess the prevalence of fibromyalgia in primary Sjogren's syndrome and to evaluate the clinical differences between patients affected with both primary fibromyalgia and primary Sjogren's syndrome and those affected only with primary fibromyalgia. METHODS: Clinical features of fibromyalgia were evaluated in 100 consecutive outpatients with primary Sjogren's syndrome and, as controls, in 90 patients with non-insulin-dependent diabetes mellitus, in 75 patients with primary fibromyalgia and in 30 healthy subjects. RESULTS: Fibromyalgia was recorded in 22% of patients with primary Sjogren's syndrome, in 12.2% with diabetes and in 3.3% of healthy controls. In the primary Sjogren's syndrome group the prevalence was significantly higher than in healthy controls (P < 0.01), but not significantly different than in diabetes. Moreover, primary Sjogren's syndrome with fibromyalgia and primary fibromyalgia patients did not differ with respect to the number of tender points, while the mean pain threshold was lower in the latter (P = 0.05). Purpura, hypergammaglobulinemia, rheumatoid factor, and a focus score > or = 1 on lip biopsy were significantly more frequent in primary Sjogren's syndrome patients without than with fibromyalgia. CONCLUSIONS: As recently reported by other authors, our study confirms the moderate increase of fibromyalgia prevalence in primary Sjogren's syndrome. Typical fibromyalgic findings are quite similar to those of primary fibromyalgia, but surprisingly, primary Sjogren's syndrome patients with fibromyalgia show a less severe global involvement than those with primary Sjogren's syndrome alone. PMID- 11858358 TI - Contribution of digit joint aspiration to the diagnosis of rheumatic diseases. AB - OBJECTIVES: To assess the diagnostic usefulness of digit joint aspiration in patients with involvement of one or more digit joints. PATIENTS AND METHODS: All patients who underwent digit joint aspiration during a rheumatology outpatient visit between 1994 and 1998 were included. Fine needles (25 or 28 G) were used, and the aspirate was immediately examined under the microscope to determine the leukocyte count and to look for crystals under polarized light. RESULTS: Digit aspiration yielded a diagnosis of crystal deposition arthropathy in 20 (29%) of the 71 patients (gout, N = 14; calcium pyrophosphate dihydrate crystal deposition disease, N = 6). In 28 patients, the amount of fluid was too small to allow analysis. The leukocyte count indicated inflammatory disease in eight of the remaining 23 patients and mechanical disease in 15. CONCLUSION: Fine-needle aspiration of symptomatic digit joints provided the diagnosis in two-thirds of patients. PMID- 11858359 TI - Myeloma and monoclonal gammopathy of uncertain significance associated with acquired von Willebrand's syndrome. Seven new cases with a literature review. AB - OBJECTIVES: Acquired von Willebrand's syndrome (AvWS) is an uncommon complication of monoclonal gammopathy of uncertain significance (MGUS) or myeloma. We investigated clinical and laboratory test abnormalities, pathophysiological hypotheses, and treatment options in this poorly known condition. PATIENTS: Five patients with MGUS and two with myeloma who met classic criteria for acquired von Willebrand's syndrome were included in this retrospective study. RESULTS: Acquired von Willebrand's syndrome was diagnosed before the gammopathy in five of the seven patients. The severity of the bleeding events was chiefly dependent on the degree of von Willebrand's factor deficiency and on the presence or absence of gastrointestinal tract angiodysplasia. Bleeding event severity was similar in patients with nonmalignant and malignant disease. An antibody that inhibited von Willebrand's factor was detected in all seven patients. Clotting returned to normal after treatment of the malignancy in one of the two patients with myeloma. In patients with MGUS, treatment is warranted only when bleeding occurs or before surgery. Von Willebrand's factor concentrates were of limited efficacy because of their short half-life. Intravenous immunoglobulins had a longer-lasting effect (about 3 weeks); this treatment was used on a regular basis in two patients with recurrent bleeding. CONCLUSIONS: A diagnosis of von Willebrand's syndrome in adulthood should prompt a search for a monoclonal gammopathy. In patients with gammopathies, simple clotting tests ensure the diagnosis of acquired von Willebrand's syndrome. PMID- 11858360 TI - Pulmonary silicosis and systemic lupus erythematosus in men: a report of two cases. AB - We reporttwo cases of coexistence of pulmonary silicosis and systemic lupus erythematosus (SLE). The patients are two men with SLE exposed to silica for 20 years. The hypothesis that silica exposure is linked to a wide variety of known or suspected autoimmune diseases, including SLE, has been discussed in the last decade but few cases of pulmonary silicosis and SLE were reported. Our purpose was to bring attention to the increasing evidence that silica may also cause or stimulate SLE, and to suggest that the researchers look for occupational exposure, mainly in male SLE patients. PMID- 11858361 TI - Dermatomyositis sine myositis and antisynthetase syndrome. AB - We describe a 66-year-old woman with cutaneous lesions typical of classic dermatomyositis, clinical evidence of antisynthetase syndrome (arthritis, Raynaud's phenomenon, mechanic's hands and interstitial lung disease with anti-Jo 1 autoantibody) and lack of muscle disease after a full muscle evaluation that included clinical, enzymatic, electromyographic, magnetic resonance imaging and histological studies. The patient did not develop myositis after 9 years of clinical disease. The association of dermatomyositis sine myositis with antisynthetase antibodies suggests that the characteristic skin lesions are closely linked with dermatomyositis on the basis of the similar clinical and serological features of both dermatomyositis sine myositis and classic dermatomyositis. PMID- 11858362 TI - Can ischemic hip disease cause rapidly destructive hip osteoarthritis? A case report. AB - Avascular osteonecrosis of the femoral head (AONFH) usually goes through the four stages described by Arlet and Ficat: normal radiographs, heterogeneity and sclerosis of the femoral head, subchondral fracture with an individualized sequestrum, and secondary osteoarthritis. Arlet and Ficat individualized a specific pattern of AONFH which they called ischemic hip disease, in which cartilage damage seen as concentric joint space loss precedes the bony alterations. Although radiological and pathological studies of ischemic hip disease have been published, no clinical data are available. We report the case of a 65-year-old man admitted for a 1-month history of severe hip symptoms with concentric joint space loss but no osteophytes. Laboratory tests and examination of fluid aspirated from the hip ruled out septic arthritis and inflammatory hip disease. Two magnetic resonance imaging (MRI) studies done 1 month apart showed diffuse edema involving not only the femoral head but also the neck and trochanter, as well as major synovial hypertrophy. This atypical MRI appearance prompted synovial membrane and pertrochanteric core biopsies, which showed reactive synovitis and stage IV osteonecrosis, respectively. The pain, disability, and joint space loss worsened. Total hip arthroplasty was performed 1 month after the biopsy. Histological examination of the femoral head showed diffuse necrosis; no evidence of another condition was found on histological sections of the entire synovial membrane. This case corroborates the hypotheses put forward by Lequesne that some cases of rapidly destructive hip osteoarthritis may be ascribable to ischemia. PMID- 11858363 TI - Joint injections in patients on antiplatelet or anticoagulant therapy: risk minimization. PMID- 11858364 TI - Giant bone islands: a case with 31 years of follow-up. AB - A dense bone lesion raises immediate concern about the possibility of a primary or secondary tumor. Stability of the image over time is a strong argument against a malignancy. Benign solitary bone islands are usually believed to remain stable over time, with no tendency toward growth. We report a case in a patient with 31 years of follow-up, during which marked growth of the island was documented. The bone scan was normal, and histology established the diagnosis of benign bone island. In a patient with an enlarging sclerotic lesion, typical radiographic features and absence of radionuclide uptake are strong arguments in favor of a bone island. PMID- 11858365 TI - Three-year outcome in a patient with Staphylococcus lugdunensis discitis. AB - The few reported cases of bone and joint infection by Staphylococcus lugdunensis indicate that the clinical manifestations are severe, the diagnosis elusive, and the treatment difficult. We report a case of lumbar discitis caused by Staphylococcus lugdunensis in a 67-year-old man receiving chemotherapy for stage III IgA lambda multiple myeloma. Treatment was with ofloxacin and pristinamycin for 1 year. Although he started to improve only 5 months after treatment initiation, the outcome was favorable. Follow-up at the time of this writing is 3 years. PMID- 11858366 TI - Hypercalcemia in a patient with tuberculous adrenal insufficiency. AB - OBJECTIVES: To raise awareness of hypercalcemia as a rare and at times inaugural manifestation of adrenal insufficiency. CASE REPORT: Evaluation of hypercalcemia in a 43-year-old man showed adrenal insufficiency. Biopsies of the testes and adrenal glands revealed epithelioid and giant cell lesions indicating tuberculosis. Although tuberculosis can contribute to hypercalcemia, this possibility was ruled out in our patient by the low serum 1,25-dihydroxy-vitamin D3 levels and return to normal of serum calcium and renal function under hormone replacement therapy. It should be noted, however, that a course of pamidronate was given. CONCLUSION: The mechanism of hypercalcemia associated with adrenal insufficiency is controversial. Hyperparathyroidism was ruled out in our patient. Adrenal insufficiency should be considered in some patients with hypercalcemia. PMID- 11858367 TI - Amyotrophic lateral sclerosis in a patient with Wegener's granulomatosis. PMID- 11858368 TI - Pseudo-septic meningeal reaction after intradural glucocorticoid therapy for sciatica. PMID- 11858369 TI - Ankylosing spondylitis and Behcet's disease in combination. Two case reports. PMID- 11858370 TI - Concerning "The fuzzy nosology of early rheumatoid arthritis and early spondyloarthropathies: square classifications produced by circular reasoning?". PMID- 11858371 TI - The effect of short-course high-dose methylprednisolone on peripheral blood CD34+ progenitor cells of children with acute leukemia during remission induction therapy. AB - This study was undertaken to determine the effect of short-course high-dose methylprednisolone (HDMP) treatment on peripheral blood (PB) CD34+ progenitor cells during remission induction treatment in 11 children with newly diagnosed acute leukemia (7 with ALL, 4 with AML) whose bone marrow (BM) cells expressed fewer than 5% CD34 at the time of diagnosis. All children who had no infection were given HDMP as a single daily oral dose of 30 mg/kg for the first four days of induction therapy. The number of CD34+ progenitor cells were determined by flow cytometry before and after four days of HDMP treatment. While the number of PB blast cells significantly decreased after only a four-day course of HDMP treatment, the number of PB CD34+ progenitor cells increased in all patients. In addition, after four days of HDMP treatment polymorphonuclear leukocytes (PMN) and mononuclear cells (MNC) increased significantly (p < 0.05). We suggest that the potential beneficial effects of HDMP in the induction treatment of acute leukemia may occur partly by the stimulation of PB CD34+ hematopoietic progenitor cells in a short period of time. PMID- 11858372 TI - Biochemical and molecular analysis of mucopolysaccharidoses in Turkey. AB - The mucopolysaccharidoses (MPSs) are a family of heritable disorders caused by deficiency of lysosomal enzymes needed to degrade glycosaminoglycans (GAGs). The undegraded or partially degraded GAGs are stored in lysosomes and/or excreted in urine. In our study, 118 patients seen over the past 20 years and suspected to have lysosomal storage disorders (LSDs) were subjected to clinical and biochemical analysis at Hacettepe University Children's Hospital. We analyzed urine and blood samples from 42 patients given a clinical MPS diagnosis. Using urine screening technique, we were able to show that 34 of the 42 patients had MPS condition. Further analysis of eight patients with normal urine MPS patterns revealed four patients as likely to have alpha-mannosidosis, fucosidosis, sialidosis, and aspartylglucosaminuria (one each). Four patients had normal oligosaccharide patterns. We were able to clearly identify 4 MPS I, 2 MPS II, 5 MPS IIIA, 8 MPS IIIB, 11 MPS IVA, 3 MPS VI, and 1 MPS IIIC patients. These results provided biochemical diagnosis for these 34 patients, and clearly show that Turkey has a higher incidence of MPS IVA, IIIB, and IIIA than of previously suspected MPS types. Molecular analysis of four MPS I patients revealed three polymorphisms which have been previously reported (A314, T388, and A461T). In MPS II patients, mutation analysis identified one previously detected (R172X) and one novel mutation (W109C). PMID- 11858373 TI - The prevalence and molecular basis of beta-thalassemia in Isparta province and region. AB - The prevalence and molecular basis of beta-thalassemia in the district of Isparta were determined in a total of 6,054 healthy high school students who were recruited from 21 randomly selected high schools in the Isparta province and region. In 182 subjects, naked eye single tube red cell osmotic fragility test (NESTROFT test) was positive HbA2 was measured by high-performance liquid chromatography (HPLC) in these subjects and was found to be high in 149 subjects. The incidence of beta-thalassemia was 149 in 6,054 (25%). The beta-thalassemia frequency was lower in the city center than in neighboring towns, 1.7% vs. 2.2%, respectively The most prevalent mutation of beta-thalassemia in this region was IVS 1-110 (G-A), followed by Codon 39 (C-T) and IVSII-745 (C-G). PMID- 11858374 TI - Analysis of thalassemia syndromes and abnormal hemoglobins in patients from the Aegean region of Turkey. AB - Turkey is located in a geographic area of the world where thalassemia syndromes and abnormal hemoglobins are common. In this study we aimed to evaluate the thalassemia syndromes and abnormal hemoglobins in patients from the Aegean region of Turkey. Among the patients admitted to our Pediatric Hematology or Hematology Clinic between January 1997-September 1999, hemoglobin electrophoresis of 3,228 cases investigated for anemia was done using high performance liquid chromatography. Beta thalassemia trait was diagnosed in 21.1%, beta thalassemia major in 0.2%, S-beta thalassemia in 0.37%, Hb D in 0.37%, Hb S trait in 0.32%, Hb E in 0.18%, Hb O-Arab in 0.12%, Hb G-Copenhagen in 0.09%, Hb D-Iran in 0.06%, Hb Lepore in 0.06%, Hb Hasharon in 0.03%. Our results demonstrate that people in the Aegean region of Turkey have a wide spectrum of thalassemia syndromes and abnormal hemoglobins. PMID- 11858375 TI - The effect of hand splints on stereotypic hand behavior in Rett's syndrome. AB - The purpose of this study was to examine the effect of hand splints and one elbow restraint on persistent stereotypic hand movements of four girls with Rett's syndrome. Among the most characteristic features of Rett's syndrome are stereotypic hand wringing and loss of previously acquired functional hand skills. Hand splints and one elbow restraint were used in this study. The subject's stereotypic hand behavior and functional hand use were calculated from five minute segmental video tape recordings. The study consisted of three phases: baseline, intervention, and withdrawal. All subjects demonstrated a decrease in stereotypic hand behavior after the application of hand splints. Although splints showed a positive effect on hand movements in Rett's syndrome, they could also lead to other, undesirable, movements. Whether splints have a positive effect on the functional use of the hand should be investigated in more subjects. PMID- 11858376 TI - Factors influencing breastfeeding for working mothers. AB - In order to evaluate the relation between breastfeeding and working conditions a descriptive study was conducted on 301 working mothers. Seventy-seven percent of mothers breastfed their infants four months and longer, and the mean breastfeeding period was 6.2 +/- 3.4 months. Forty-one percent of mothers started weaning before four months of age. The multifactorial analysis of independent factors significantly influencing breastfeeding time were, in decreasing order of significance, breastfeeding conditions at work, maternal leave period, mother's smoking habit and the use of breastpump. For weaning period, these factors were the use of breastpump, breastfeeding leave at work and maternal leave period. In conclusion, in order to support breastfeeding at work, maternal leave period must be prolonged, and breastfeeding conditions at work must be improved. PMID- 11858377 TI - Nutritional status of children with cancer and its effects on survival. AB - In the present study we aimed to determine the nutritional status of our patients and to assess its relationship with survival. The nutritional status of 47 patients with cancer was evaluated at diagnosis, three months after initiation of the treatment and at the end of therapy. Weight for height, height for age, and weight for age of children were expressed as percent of standard. Values for each nutritional index were converted into standard deviation (Z) scores. Three-year overall survival (OS) and event-free survival (EFS) rates of patients were determined according to their nutritional status. The overall prevalence of malnutrition at diagnosis was 29.8%. Three months later the malnutrition ratio reached 38.3% and then decreased again to 18.5% at the end of the therapy. Although the prevalence of malnutrition at the third month of treatment was significantly higher from the prevalence at diagnosis (p: 0.001) and at the end of the therapy (p: 0.009), the mean Z scores of the nutritional indexes before and during the treatment were not significantly different. The survival rates of malnourished patients were not different from those of well nourished patients. In conclusion, malnutrition is one of the main problems in children with cancer; hovewer, nutritional status has no effect on survival. PMID- 11858378 TI - The effects of gemfibrozil on hyperlipidemia in children with persistent nephrotic syndrome. AB - Persistent nephrotic syndrome is frequently accompanied by severe hyperlipidemia, and this may pose a substantial risk for cardiovascular disease. Lipid-lowering drugs are prescribed by many nephrologists for adult patients but rarely for nephrotic children. The present investigation was designed to evaluate the safety and efficacy of gemfibrozil in nephrotic children. Eight girls and four boys aged from 5 to 17 years were enrolled in this study. They were all steroid and immunosuppressive resistant patients with nephrotic range proteinuria. Placebo was administered to five patients and gemfibrozil was administered to seven patients for four months. Blood samples were taken for the determination of cholesterol, triglyceride, low-density lipoprotein (LDL), high-density lipoprotein (HDL), BUN, serum creatinine (Scr), ALT, AST, CPK, apolipoprotein A (apo A), apoliporotein B (apo B), and serum albumin levels during the initial and subsequent examinations. At the end of the fourth month, gemfibrozil reduced total cholesterol by 34%, LDL by 30%, apo B by 21% and triglycerides by 53% (p < 0.05). HDL cholesterol and apo A levels were not significantly altered. Renal function and urine protein excretion were not affected by gemfibrozil. In this study gemfibrozil therapy had no side effects and had favorable effects on the lipoprotein profile of nephrotic patients. PMID- 11858379 TI - Meningitis due to Salmonella in preterm neonates. AB - Meningitis due to Salmonella is a rare condition. Here we report seven cases with neonatal Salmonella meningitis diagnosed and treated in the Pediatric Neonatology Unit of Ondokuz Mayis University Faculty of Medicine between January 1985 February 2001. Five cases were cured with appropriate treatment; in two of them no neurological sequelae were seen. Of the remaining three, reversible hydrocephalus occurred in two cases and subdural empyema was found in one. Two patients died on the 30th and 40th days of hospitalization: one of them had hydrocephalus and the other ventriculitis. In this report we point out the importance of neonatal meningitis due to Salmonella serotypes, though it rarely occurs. PMID- 11858380 TI - Complications and outcome in left-sided endocarditis in children. AB - We retrospectively assessed the clinical course and outcome of left-sided endocarditis in pediatric patients to find out the prognostic significance of the presence and size of echocardiographically detected vegetations. Among the children admitted to our institution with endocarditis between January 1987 and October 1999, 16 patients (mean age 9.03 +/- 4.95 years) who met the Duke criteria for the diagnosis of infective endocarditis (IE) were included in this study. Rheumatic valvular disease was the most frequent underlying heart disease (10 patients: 62.5%). Five patients were operated at a mean of 13.9 months before endocarditis, and all had residual defects. Vegetation was detected in 11 cases (69%). Ten patients had major complications (within 2 weeks in 6 patients). Three patients developed congestive heart failure (CHF), six had intracranial and one had lower extremity emboli. Among them four were operated because of complications (CHF: 3 cases, intracranial emboli: 1 case). All the operated cases are doing well. The association between intracranial embolic events and echocardiographically detected vegetations was determined by calculating specificity (40%), sensitivity (100%), positive predictive value (50%), and negative predictive value (100%). No intracranial embolism occurred in patients without vegetations. All vegetations were < or = 6 mm in patients with systemic embolism. There were four deaths, three of which were because of intracranial embolism. This study suggests that intracranial emboli have a major risk of mortality in left-sided endocarditis. The larger size of the vegetation is not a predictor of complications; furthermore, the absence of vegetations predicts that the patient is safe from embolic events. Therefore all patients with left-sided IE should be considered for earlier surgical intervention. PMID- 11858382 TI - Lamellar ichthyosis: a case report. AB - Ichthyoses are divided into four groups according to clinical, histopathologic and genetic findings. Lamellar ichthyosis is one of them. The incidence of lamellar ichthyosis is believed to be approximately 1 per 100,000 to 300,000 live births. It is characterized by large, polygonal, grayish brown, and tightly adherent scales. We report a four-year-old boy with desquamative lesions since birth who had six-year-old sister with similar lesions, suggesting an autosomal recessive inheritance. His skin biopsy revealed hyperkeratosis with lamellae. There were no associated hair or neurological abnormalities. His clinical and histopathological findings were typical for isolated lamellar ichthyosis. Because of its rare occurrence, we report this case with a review of the literature. PMID- 11858383 TI - Oral acitretin treatment in severe congenital ichthyosis of the neonate. AB - Two newborn infants with ichthyosis, one with lamellar ichthyosis and one with nonbullous ichthyosis form erythroderma, who presented at birth with a collodion baby appearance, were treated with acitretin (1 mg/kg/day). Clinical improvement was achieved shortly after treatment. The second case received oral retinoid for 3.5 months and was followed for nine months. The result was excellent. The treatment resulted in a satisfactory improvement in the skin condition of the first case. The tolerance to the drug was good. Side effects were not observed. It was concluded that early management of severe ichthyosis cases could prevent life-threatening events such as hyperthermia, disturbance in electrolyte and fluid balance, and infection. PMID- 11858381 TI - Acute isoniazid neurotoxicity in childhood. AB - Acute isoniazid (INH) poisoning is uncommon in children. Although most physicians are aware of INH hepatotoxicity, acute INH poisoning and its treatment are not well recognized. INH is increasingly being used to control the spread of tuberculosis, and physicians should know its potentially fatal effects. INH overdose is known to result in rapid onset of seizures, metabolic acidosis and prolonged obtundation. We report two cases of obtundation secondary to INH overdose that was immediately reversed by pyridoxine. Parenteral pyridoxine administration is an effective method in INH intoxication. The intravenous form of pyridoxine must be available in the emergency care units, and INH toxicity should be suspected in any patient with refractory seizures and metabolic acidosis. PMID- 11858384 TI - A huge gastric stromal tumor in a 13-year-old girl. AB - A 13-year-old girl presenting with severe anemia was diagnosed to have a large gastric tumor protruding toward the antrum with two central ulcerations. Partial gastrectomy including antrectomy and gastroduodenostomy were performed. Histologic and immunohistochemical studies revealed one of the most uncommon gastric tumors in children; a gastrointestinal stromal tumor. Close follow-up of the patient with endoscopy, abdominal ultrasonography and/or computed tomography in three to six month intervals revealed no recurrences or metastasis of the tumor following its complete excision. PMID- 11858385 TI - Familial secundum atrial septal defect with dysrhythmia associated with web neck. AB - Most cases of atrial septal defect occur sporadically, but a few families have the defect as a genetic abnormality. A family having familial type secundum atrial septal defect with dysrhythmia associated with web neck is reported. In this family, two female siblings aged 11 (Case 1) and 4 years (Case 2) and their father had secundum atrial septal defect. Case 1 presented with two year history of syncope attacks and Case 2 with easy fatigability since early childhood. Both sisters also had web neck as a solitary anomaly. Electrocardiograms revealed prolonged PR interval and right bundle-branch block in both cases. In Case 1 first-degree atrioventricular block and Mobitz type I and II block were observed in Holter monitoring. Echocardiographical examination showed secundum atrial septal defect in both sisters. A permanent pacemaker was implanted in Case 1, and then atrial septal defects in both patients were surgically repaired; no postoperative complaints were observed. The father had been diagnosed as having atrial septal defect when he was 35 years old, and first-degree atrioventricular block and atrial flutter developed after open heart surgery. In conclusion, the association of secundum atrial septal defect and prolongation of PR interval should be considered as familial occurrence of atrial septal defect. Identification of atrial septal defect in more than one family member should prompt clinical evaluation of all relatives. PMID- 11858386 TI - Peripheral facial paralysis as initial manifestation of hypertension in a child. AB - Hypertension is one of the rare causes of peripheral facial paralysis in children. The unawareness of this association at presentation may cause serious medical errors and result in delays in the diagnosis of hypertension, which may worsen with corticosteroid therapy given for Bell's palsy. We describe a severely hypertensive child who was first seen with peripheral facial paralysis and given corticosteroid therapy in another hospital. She presented to our clinic during the second facial paralysis attack with hypertensive pontine hemorrhage. PMID- 11858387 TI - Congenital mediastinal immature teratoma: a case report with autopsy findings. AB - A full-term newborn with karyotype 46, XX was delivered by cesarean section. She had severe respiratory distress and substernal retraction, and underwent emergency operation, but she died on the same day due to respiratory failure. The mother, 26-year-old prima gravida with no history of twinning, had been examined with ultrasonography at the 34th week of her pregnancy, which revealed a fetus with edema of head and neck region, a probable diaphragmatic hernia, polyhydramnios, and a large mediastinal mass with solid and multicystic parts with hypoplasia of the lungs. Autopsy revealed a 9 x 5 x 3 cm lobulated mediastinal mass with both solid and cystic areas, displacing the lungs and the heart postero-inferiorly and thymus anteriorly. The lungs were hypoplasic. Microscopically, the mass showed mature epithelial and mesenchymal tissues with primitive mesenchyme and immature neuroepithelium. All these findings led to the diagnosis of an immature teratoma. Mediastinal teratomas are rare and life threatening, but early diagnosis and surgical intervention in a newborn with sufficient lung maturation may provide a long survival. PMID- 11858388 TI - An unusual case of esophageal and laryngotracheal atresia. AB - Esophageal atresia with or without tracheoesophageal fistula is a relatively common congenital anomaly. However, esophageal atresia with associated laryngotracheal atresia, double tracheoesophageal fistula and cardiac malformations is an extremely rare condition. In this article we report a newborn infant with severe respiratory distress at birth who had both esophageal and laryngotracheal atresia with congenital cardiac malformations, in an attempt to bring attention to the clinical presentation, and emergent diagnostic and therapeutic approaches. PMID- 11858389 TI - Isolated noncompaction of the ventricular myocardium. AB - Noncompaction of the ventricular myocardium is a rare congenital disorder characterized by the presence of numerous prominent trabeculations and deep intertrabecular recessess which communicate with the left ventricular cavity. The disease uniformly affects the left ventricle, and sometimes also affects the right ventricle. Echocardiographic findings are important clues for the diagnosis. Clinical symptoms include signs of left ventricular systolic dysfunction even to the point of heart failure, ventricular arrhythmias, and embolic events. We describe an illustrative case of isolated noncompaction of the left ventricular myocardium in a two-year-old child with the typical clinical and echocardiographic features of the disease. The literature on the topic is reviewed. PMID- 11858390 TI - The second case with 47, XY, + 8 [38] / 45, X0. PMID- 11858391 TI - Femoral reconstruction. AB - Reconstruction of the femur is sometimes difficult in patients that may sustain long-term disabilities. Following advancements in modern technology, some femoral disorders which were considered to be recalcitrant in the past can now be treated with satisfactory outcomes. This article reviews the experience of femoral reconstruction at the Orthopedic Department of Chang Gung Memorial Hospital (Linkou). Reconstruction of the femur includes nonunion, malunion, and segment bony defects that require bone transfer. The prerequisite for successful treatment was to follow the biological and biomechanical principles. The author believes that the technique of femoral reconstruction can be developed and simplified so that success rates can be further improved. PMID- 11858392 TI - Lateralization and prognostic value of proton magnetic resonance spectroscopy in patients with intractable temporal lobe epilepsy. AB - BACKGROUND: The aims of this prospective study were to investigate the stability of hippocampal metabolite ratios obtained by proton magnetic resonance spectroscopy (MRS), to evaluate the ability of MRS to determine the pre-surgical lateralization of seizure focus, and to assess the relationship between MRS results and postoperative outcomes in patients with intractable temporal lobe epilepsy. METHODS: Within- and between-acquisition variations were evaluated in 30 control subjects, using the chemical-shift imaging technique. The most stable metabolite ratio was then applied for pre-surgical evaluation of 14 patients with intractable temporal lobe epilepsy. RESULTS: The ratio between N-acetylaspartate (NAA) and choline-containing compounds (Cho) plus creatine-phosphocreatine (Cr), i.e., NAA/(Cho+Cr), had an overall smaller percentage change between measurements (13%-28%) than did the other ratios: NAA/Cho (18%-37%), NAA/Cr (11%-60%), and Cho/Cr (19%-51%). With a mean follow-up period of 27 months (range, 12-55 months) after an anterior temporal lobectomy, 10 patients were in Engel s class I (71%), 2 were in class II (14%), and 2 were in class III (14%). Lateralization by MRS was consistent with the operation side in 11 (79%) patients, including class I in 8 patients, class II in 1 patient, and class III in 2 patients. Ranges of reduced NAA/(Cho+Cr) values were similar between patients with different post-surgical outcomes: 0.26-0.47 for class I, 0.40-0.45 for class II, and 0.34-0.40 for class III (p=0.651). CONCLUSIONS: Proton MRS may be helpful in pre-surgical lateralization of the seizure focus in patients with intractable temporal lobe epilepsy; however, it cannot provide prognostic information about postoperative seizure control. PMID- 11858393 TI - Implications of sonographic identification of duplex kidney in adults. AB - BACKGROUND: Duplication of the renal pelvis and ureter constitutes the most common anomalies of the upper urinary tract. Most patients are asymptomatic and don't need treatment. However, the associated anomaly and acquired renal disease may cause morbidity and mortality. The aim of this study was to investigate the prevalence of duplex kidney in sonographic examinations and the predictive capability of this method. An analysis of patients with urography-confirmed duplication anomaly was also performed. METHODS: During a period of 9 year (from 1988 to 1996), sonographic examination records of 19,287 patients were reviewed. Patients with a diagnosis of duplex kidney were enrolled. Their clinical characteristics and radiological images were recorded and evaluated. RESULTS: One hundred and sixty-three patients were found by sonography to have duplex kidney, for an prevalence of 0.85%. Urography studies were done on 64 (39.2%) patients, and 42 patients were proven to have duplex kidney. The positive predictive rate of sonography was 65.6%. Eighty percent of patients with duplex kidney had associated ureteral duplication. No associated anomalies were found. Four patients had urolithiasis, and 3 patients had recurrent urinary tract infection. One patient had coexisting uroepithelial malignancy, and 1 patient had refractory gross hematuria. CONCLUSIONS: Duplex kidney is an uncommon finding during adult sonographic examinations. The sensitivity and specificity of sonography in identifying this anomaly are controversial. An experienced hand with good knowledge of the anomaly and its sonographic signs is helpful in more accurately diagnosing this anomaly. PMID- 11858394 TI - Lens subluxation: 10-years' experience at Chang Gung Memorial Hospital, Kaohsiung. AB - BACKGROUND: This study aimed to describe the clinical manifestations and surgical outcomes of subluxated lens. METHODS: Reviewing charts from January 1991 to June 2000, we studied 110 patients (123 eyes) who had lens subluxation at Chang Gung Memorial Hospital, Kaohsiung. For 21 eyes, patients accepted conservative treatment, and 102 eyes underwent surgery (8 eyes had Marfan syndrome; 18 eyes had idiopathic dislocation; and 76 eyes had experienced trauma). Indications of surgery included reduction in visual acuity, uncorrectable refractive error, and secondary glaucoma. RESULTS: There were 81 male and 29 female patients. Their mean age was 47.6 years (range, 2-76 years). The mean post-operative follow-up period was 22.24 months (range, 2-118 months). In spite of the different lens statuses, similar visual results were found between the surgical treatment group and the nonsurgery group. In the surgery group, 47 eyes (46.1%) achieved best corrected visual acuity (BCVA) of 20/40 or better. A total of 80 eyes (78.4%) achieved an improvement (log MAR change >0.3) in BCVA. The major postoperative complications included glaucoma (11 eyes), retinal detachment (9 eyes) and intraocular lens dislocation into the vitreous cavity (3 eyes). CONCLUSIONS: Results indicate that the presence of a subluxated lens alone is not an absolute indication for surgical removal. The lens status had no significant influence on surgical outcome. Most cases can achieve an improvement in BCVA after appropriate management. PMID- 11858395 TI - Measurement of serum vitamin E isomers in fullterm and preterm infants. AB - BACKGROUND: This study was conducted with the following aims: (1) to construct a methodology by high-performance liquid chromatography (HPLC) to measure vitamin E levels in our neonatal units; (2) to establish a reference range of serum vitamin E levels in fullterm neonates and their mothers; and (3) to compare these to levels in the preterm group. METHODS: During a 6-month period from September 1996 to February 1997, cord blood of 34 uneventful fullterm neonates (gestational age 38-42 weeks, birth weight 2840-3530 g) and venous blood of their healthy mothers were collected. Venous blood of 35 preterm neonates (gestational age 28-34 weeks, birth weight 940-1980 g) was collected at the same time. Serum vitamin E levels were measured by HPLC. The serum vitamin E/total lipids ratio was also examined. RESULTS: Serum levels of alpha-tocopherol in 34 fullterm neonates and their mothers were 0.212+/-0.127 and 1.160+/-0.513 mg/dl, and of gamma-tocopherol were 0.029+/-0.019 and 0.214+/-0.122 mg/dl, respectively. All fullterm neonates except 2 had serum levels less than 0.5 mg/dl. There was a positive relationship between serum E levels and total lipids. In addition, the correlation between neonatal vitamin E/total lipids ratios and maternal vitamin E/total lipids ratio was statistically significant. The serum level of alpha-tocopherol in 35 preterm infants was 0.170+/-0.090 mg/dl and that of gamma-tocopherol was 0.020+/-0.015 mg/dl; both levels were much lower than those of their fullterm counterparts. But the vitamin E/total lipids ratios did not significantly differ. CONCLUSIONS: We conclude that (1) measurement of serum vitamin E levels by the HPLC method is practical, and is useful in evaluating deficiencies in preterm infants and possibly useful in monitoring the adequacy after supplementation in future study, and (2) preterm infants had a much lower level of serum tocopherol, but no difference was seen in vitamin E/total lipids ratio. PMID- 11858396 TI - Reduction of subretinal fluid after preoperative immobilization of the eyes with rhegmatogenous retinal detachment. AB - BACKGROUND: Bullous retinal detachment may be difficult to handle with higher operative complications. Preoperative immobilization of the eyes might reduce subretinal fluid and facilitate surgical procedures. Factors influencing subretinal fluid absorption were analyzed. METHODS: Twenty-eight eyes with primary rhegmatogenous retinal detachment with preoperative binocular patching and complete bed rest for at least 12 hours were enrolled. Ultrasonography was performed before and after immobilization to estimate the reduction in subretinal fluid. RESULTS: The amount of reduction diverged in cases with an average of a 17.1% decrease in the maximal height of detachment. Retinal detachments in patients with smaller breaks and without vitreal traction or chronicity showed more significant reduction of subretinal fluid than did those patients with larger breaks and with vitreal traction or chronicity. On the other hand, break shape, height of retinal detachment, age, and gender had no obvious effect on subretinal fluid change. CONCLUSIONS: Preoperative immobilization of the eyes reduces the subretinal fluid in selected cases, which can decrease operative complications. Break size, vitreal traction, and chronicity are important factors affecting the absorption of subretinal fluid. PMID- 11858397 TI - Torsade de pointes induced by metoclopramide in an elderly woman with preexisting complete left bundle branch block. AB - There is a growing list of drugs implicated in acquired long QT syndrome and torsade de pointes. However, the torsadogenic potential of metoclopramide, a commonly used antiemetic and prokinetic drug, has not been reported in the literature, despite its chemical similarity to procainamide. We report on a 92 year-old woman with preexisting complete left bundle branch block who developed torsade de pointes after intravenous and oral administration of metoclopramide. This patient also developed torsade de pointes when cisapride and erythromycin were given simultaneously. These two episodes were suppressed successfully after discontinuing the offending drugs and administering class IB drugs. This is the first documentation that metoclopramide provokes torsade de pointes clinically. Metoclopramide should be used cautiously in patients with a risk of torsade de pointes. PMID- 11858398 TI - Candida vertebral osteomyelitis: a case report and review of the literature. AB - Candida species are low virulence organisms which inhabit the skin and mucous membranes of most individuals. There has been increasing incidence of disseminated and deep-seated Candida infections owing to the increasing number of immune compromised hosts. However, the Candida species are still rarely suggested as causative pathogens of vertebral osteomyelitis. We present a 51-year-old man with neck pain and cervical radiculopathy. Three months prior to visiting our hospital, he had undergone a urological operation which was complicated by a urinary tract infection. Magnetic resonance imaging and X-rays showed erosion of the body of the 5th cervical spine and collapse of the C5-C6 disc. After open debridement, tissue pathology results revealed Candida infection. Clinical stability was achieved during the 6-month follow-up period with a combination of amphotericin B and posterior fixation method. We reviewed the literature and found a high rate of surgical intervention for patients suggested of having Candida vertebral osteomyelitis. However, the surgical intervention may not be necessary. Early diagnosis using noninvasive percutaneous needle biopsy may help reduce the incidence of delayed treatment. PMID- 11858399 TI - Isolated craniosynostosis: prenatal ultrasound of scaphocephaly with polyhydramnios. AB - Craniosynostosis is an abnormal shape or dimension of the skull caused by premature closure of one or more skull sutures. It includes scaphocephaly, brachycephaly, oxycephaly, plagiocephaly, trigonocephaly, turricephaly, and a cloverleaf-shaped head. The only cases of craniosynostosis that have been reported concern fetuses with complex and marked craniosynostosis syndromes such as acrocraniofacial dysostosis, Apert's syndrome, Beare-Stevenson cutis gyratum syndrome, Calabro's syndrome, etc. Isolated or simple craniosynostosis, however, is a developmental anomaly rarely found on routine ultrasound examination. We present a case that was diagnosed as scaphocephaly with moderate polyhydramnios by prenatal ultrasound. No other structural anomaly was detected, and the karyotyping was normal. So isolated sagittal craniosynostosis was diagnosed prenatally and was confirmed by postnatal skull radiography as well as 3 dimensional computed tomography. The infant underwent bilateral parietal craniectomy at the age of 3 months. He obtained a good result with no mental problems 4 years after the operation. To our knowledge, this is the second report on the prenatal detection of an isolated form of craniosynostosis. PMID- 11858400 TI - Primary spinal epidural non-Hodgkin's lymphoma in a child. AB - Non-Hodgkin's lymphoma usually involves the central nervous system by metastatic disease. Primary spinal epidural non-Hodgkin's lymphoma (PSENL) is a relatively rare cause of spinal cord compression. It mainly occurs in adults past the 4th or 5th decades. This entity is even rarer in children. The proper treatment modalities are controversial in adults with PSENL. Radiotherapy is the main strategy after surgery; the role of chemotherapy is uncertain. Therapeutic experience in childhood PSENL is extremely limited. We report a 10-year-old boy presenting with backache and bilateral lower leg weakness after minor trauma. Small non-cleaved cell non-Hodgkin's lymphoma of the epidural space was proven after subtotal tumor removal. Other investigations including computed tomography of the chest and abdomen, bone scan, gallium scan, bone marrow aspiration, and cerebrospinal fluid study were all negative for occult disease. The patient received combined therapy with irradiation and chemotherapy after surgery. Esophageal stricture resulting from radiotherapy developed during treatment and colon interposition was performed. He has remained disease free 42 months after the diagnosis with normal functional status. PMID- 11858401 TI - Myxoma of the gingiva: a case report and literature review. AB - Soft tissue myxomas of the oral and para-oral tissue are extremely rare. Only two cases of myxoma of gingiva have been reported in the literature. The histogenesis of these lesions remains obscure. We report a case of gingival myxoma in a 37 year-old man. A firm ovoid soft tissue mass measuring 1.5 x 1.2 x 1 cm in size was noted at the mesio-lingual gingiva of a partially impacted right mandibular third molar. Radiographic examination revealed an impacted mandibular third molar with normal radiographic bony consistency of the mandible. Excisional biopsy was performed with no evidence of recurrence after 8 months. Histological examination showed stromal mass composed of myxoid-like fluid in fibrovascular stroma and scanty inflammatory cell infiltration, suggesting myxomas of the oral soft tissues and jaws. Immunohistochemical stains with S-100 protein, desmin and smooth muscle specific actin yielded negative results, and those with alcian blue, periodic-acid-Schiff, mucicarmine, vimentin and reticulin were positive. Tumor cells of mesenchymal origin without neurogenic or muscular derivations and myxoid matrix composed of acid mucopolysaccharides in this lesion further confirmed the diagnosis of myxoma. Periodontal ligament origin cannot be completely excluded due to tumor location and its association with an impacted molar. However, further study is necessary to clarify the origin and histogenesis of these lesions. PMID- 11858402 TI - Bridging the gap. Community partnerships in nursing education: advancing primary health care in practice. AB - An innovative approach to nursing education was piloted by the School of Nursing at Flinders University and Noarlunga Health Services, a generic community health service, in Adelaide, South Australia. The approach, encapsulated in the four year Community Enrichment Project (CEP), focused on Primary Health Care (PHC) and nursing practice. The CEP developed curriculum and organised student placements, which promoted understanding of PHC in a way that integrated acute and community sectors. This paper considers the impact of splitting the students' third year final practicum, of seven weeks, between acute and community placements. Integral to the overall outcome of the project was the community agencies' cooperation in accepting students for longer clinical placements than had previously been the norm. This resulted in students being engaged in activities and projects in diverse areas. The students were aided in linking PHC theory to practice by the CEP team members who mentored them throughout these placements. Outcomes demonstrated the ability of students to integrate PHC theory to practice, across a variety of health settings. Students moved from a position of little knowledge or understanding of PHC, to an integrated knowledge of PHC principles and the relationship to enhanced nursing practice. PMID- 11858403 TI - The reliability of measuring nursing clinical performance using a competency based assessment tool: a pilot study. AB - The clinical performance of the novice nurse has been identified as problematic in the areas of medication administration, time management, and comfort and safety. In order to assess new graduate nurses' clinical competence in these areas, a forty four (44) item assessment tool was developed which was drawn from the Australian Nursing Council Inc. (ANCI) (1991) competency statements. A correlation descriptive design was used to determine the inter-rater reliability between assessors using this competency based tool. A pilot study utilising a convenience sample of 30 new graduate nurses were each assessed by two observers for a mean period of 127.5 minutes. Kappa coefficients were calculated to determine the level of agreement between the two observers. Very little rating agreement was found for individual items, however, there was no significant difference in the total competency scores between the two observers. The competency assessment tool has face validity and is internally consistent. However, inter-rater reliability for individual competency items was found to be poor. PMID- 11858404 TI - A collaborative approach to developing nursing skills to meet service needs. AB - Academics, educators and health service managers came together to explore, within their local area, the nature of the perceptions of beginning registered nurses (BRNs) and Nursing Unit Managers (NUMs) to the skills utilised by BRNs within the first six months of practice. A group of 71 BRNs and nine NUMs participated in a cross-sectional survey. BRNs were also asked to rate their preparedness in each particular skill, dependent on the method of teaching--theory/lectures, nursing laboratories and clinical experience. Some 21 skills, relating to medication/fluid administration and skin and wound care, were perceived by 95 per cent or more of the graduates as being used within the first six months. Similar perceptions existed for NUMs and BRNs for 52.4 per cent of the listed skills. Differences in perceptions have led to academics altering the scope of skills being taught and focusing assessment on critical skills for BRNs, and managers increasing awareness among NUMs as to the source of the differences and developing programs to improve clinical interpretation skills. Improvements in the experience of the BRN are likely to occur when all parties work together to bring about positive change. By Barbara Somerville, PMID- 11858405 TI - Oncology decision tree. AB - Financial cutbacks and budgetary constraints continue to be a part of government policy, and impact deeply on many aspects of daily life in Australia in the late 90s. Health care is not immune from these measures, and nurses and other allied health professionals are having to manage increasingly complex patient care with less funding and resources. Patients undergoing chemotherapy and radiotherapy can usually be managed on an outpatient basis, but need round the clock specialist nursing care at home following treatment which can have unpleasant side effects. Home nursing is an expensive option, but it has been shown in the US that a specialist nurse can triage by telephone, providing expert and timely advice and care to this vulnerable group, preventing admission to hospital and providing family support. The purpose of this paper is to demonstrate that, despite budget constraints, an improved cost effective service can offer clients expert care at a fraction of the cost of alternative services. By Sally Koodiaroff. PMID- 11858406 TI - The South Australian Nurse Practitioner Project: a midwife's perspective on a new initiative. AB - Midwifery education in Australia is currently the focus of intense debate both within and outside the field of midwifery. Proposed changes arsing from these debates centre principally around the issue of midwifery as a separate profession from nursing. This paper describes a case in point as to how changes in attitude and practice are occurring. As a midwife representing the Australian College of Midwives Inc.- SA Branch (ACMI-SA) on the Advisory Committee of the South Australian Nurse Practitioner Project, I provide an insight into some of the debates around midwifery and its relationship with this project. ACMI was a member of the Ministerial Advisory Committee established following the initiative of the Executive of the Department of Human Services (DHS) (formerly the South Australian Health Commission) in forming the Nurse Practitioner Project. The Terms of Reference for this committee included the development of an operational framework for the development and implementation of the Nurse Practitioner role in South Australia. A collaborative approach was seen as essential to enable nurses to best serve their communities by functioning at an advanced level of practice. The ACMI and midwives generally fully support the Nurse Practitioner Project for nurses. By Jennifer Pauline Byrne. PMID- 11858407 TI - Secondary intention wound healing--pathphysiology and management. PMID- 11858408 TI - The electronic health record. PMID- 11858410 TI - Assessing for depression in older people. AB - Perceptions about what constitutes depression contribute to poor know edge of the condition amongst non-psychiatric medica and nurs ng staff. Stereotypes about ageing and depression go some way to explaining why depression remains under recognised and under addressed in older people. This paper highlights some of the factors that contribute to the missed or misdiagnosis of depression. It also makes explicit some of the issues surrounding diagnosis and management of the disorder. The importance of a good health history and an appreciation of the adverse effects of drugs are identified as important in assisting to diagnose this treatable condition. PMID- 11858411 TI - A day in the life of a children's nurse. AB - Practice development posts have evolved in the past decade since the demise of the Clinical Nurse Manager role. The Practice Development Adviser supports and facilitates clinical practice innovations with the aim of improving the quality of patient-focused care. Clinicians are encouraged to utilise ward-based clinical developments as an integral part of a course leading to academic qualifications. This article shares examples of clinical practice developments facilitated for a group, an individual clinician and on-going innovations. PMID- 11858412 TI - Health implications of modern childhood. AB - Childhood has undergone many reconstructions since its 'recognition' in the 18th century. The medicalized child is rooted in eugenics, political economy and professional interests and has implication for practice. The modern child is now viewed as a stakeholder in a political sense. Childhood requires recognition in a 'health' sense if it is to survive as a category. Different medical expectations and models of childhood are likely to evolve. PMID- 11858413 TI - Higher education: making an informed choice. AB - Potential students need to make informed choices when undertaking further study. Potential students should access different sorts of information regarding the institution of their choice. Potential students need to be aware of how service needs influence the choices available. Potential students should have a clear idea of what they are aiming for. Potential students should be aware of the differences between similar courses. PMID- 11858414 TI - Children's competency to consent: an ethical dilemma. AB - The application of the best interests principle in current legislation creates an ethical dilemma in relation to children's consent to treatment. The guiding principle of the UN Convention on the Rights of the Child (1989) symbolises a formal expression of children's participation rights. Children's rights to consent to treatment are granted on socially determined ideals of competency. Children's participation in health care is increasingly advocated in legislation but many barriers remain. Nurses can facilitate children's participation through communicating information and creating partnerships with children. PMID- 11858415 TI - Making the invisible, visible. AB - Special needs children experience all the difficulties experienced by any child admitted to hospital although these are hugely magnified. Parents continue to carry the burden of care, even after hospital admission, when their child has complex needs. Nurses need to remember that these children have all the needs of any child in addition to needs specific to their disability. Nurses should focus what the child can do, rather than what they cannot do. PMID- 11858416 TI - Pain at home: children's experience of tonsillectomy. AB - Earlier discharge following tonsillectomy increases the need for good pain management advice and effective analgesia. An audit determined the nature of children's pain experiences at home following tonsillectomy and identified pain management strategies used. Combined analgesia and formal pain assessment significantly reduced the number of children in moderate or severe pain on discharge. Pain could worsen following discharge, persist for three to ten days and be at times moderate to severe. 50% of parents contacted their GP and 75% of children required paracetamol and ibuprofen concurrently. Audit data was utilised to develop comprehensive written pain management advice and a discharge protocol for combined analgesia. PMID- 11858417 TI - The role of the infection control professional in the intensive care unit. AB - By design, multiple invasive procedures are performed in the intensive care unit (ICU). Although great care is taken to control morbidity and forestall mortality, this invasive environment places ICU patients and staff at immense risk of nosocomial (hospital-acquired) infection. The role of the infection control professional (ICP) within the ICU involves data collection, dissemination of data with feedback, expertise in the investigation of outbreaks, product evaluation proficiency, and fluid consultation aptitude. This article provides an inside view of how specialty infection control staff and ICU staff can optimize infection control to decrease the incidence of nosocomial infections. A description of what the ICP does not do in the ICU is also given, providing a clear guideline for how these two disciplines can best provide a safe intensive care experience. PMID- 11858418 TI - Applying social and behavioral theory as a template in containing and confining VRE. AB - Infection control professionals play several important roles--surveyors, educators, and ultimately change agents--in the identification and prevention of nosocomial infections in hospitals. The medical and surgical intensive care units (ICUs) in a large inner-city teaching hospital experienced an increased patient colonization rate with vancomycin-resistant enterococcus (VRE). Intervening in this problem required a multifaceted approach to control the spread of VRE and to change behavior by shifting social norms at multiple levels throughout the ICU community. The success of the interventions could be best explained by applying the use of several behavioral science models. The Ecological Model of Behavior Change, the Health Belief Model, and Social Cognitive Theory can be applied and are consistent with the successful interventions. This multifaceted approach to intervening in this problem consists of five levels of influence: (1) intrapersonal or individual factors, (2) interpersonal factors, (3) institutional factors, (4) community factors, and (5) public factors. We implemented educational inservices and developed references, policies, and programs directed at each of the five levels of influence. The Health Belief Model and Social Cognitive Theory were employed for intervention, and behavior change was based on modeling, observational learning, and vicarious reinforcement. Within six months of initial implementation, the number of positive VRE surveillance cultures and positive clinical isolates decreased significantly in both the medical and surgical ICUs. Two years later, there continues to be a marked reduction of VRE. PMID- 11858419 TI - Emerging multiresistant organisms in the ICU: epidemiology, risk factors, surveillance, and prevention. AB - Recent cases of vancomycin-resistant enterococcus (VRE) at our facility and the emergence of new vancomycin intermediate resistant strains of bacteria provided the impetus for this discussion on superbugs. Each year approximately two million patients develop an infection after receiving health care in a United States hospital. These infections are often difficult to treat because the microorganisms most frequently involved are resistant to current antibiotic therapy. The result is increased morbidity, mortality, and health care costs. The recent development of intermediate resistant strains of bacteria is indeed an ominous indicator of what the future might hold for the health care profession should our current practices go on unchecked. This article will present an overview of the epidemiology of these multiresistant organisms, risk factors inherent to the intensive care environment, nursing responsibilities, and the rationale for surveillance protocols. PMID- 11858420 TI - Don the barriers. AB - Health care workers (HCWs) have many "barriers" at their disposal to protect them from risk of occupational exposures to bloodborne pathogens or communicable diseases. These barriers include the daily use of personal protective attire, such as gloves, gowns, masks, and eye protection; following appropriate isolation precautions; using work practice controls, such as safety needles; using preventive methods less frequently needed, such as vaccination against the hepatitis B virus, annual influenza vaccinations, and prompt reporting of occupational exposures. Proactive and consistent use of these barriers makes any health care environment a safer place to work. PMID- 11858421 TI - Protocols for the prevention of intravascular device-related infections. AB - The frequency of nosocomial infections in intensive care unit (ICU) patients is estimated at 20%. Many of these infections are related to the frequent and prolonged use of intravascular devices. If ICU personnel are familiar with the established protocols for the selection, insertion, and maintenance of intravascular devices and are strict in their adherence to these protocols, it is expected that a significant number of these infections can be avoided. PMID- 11858422 TI - Cleaning reusable equipment in the ICU. AB - Cleaning is the single-most-important step in making a medical device safe for handling. It is also extremely important as a preparatory step before any subsequent disinfection or sterilization process. This article addresses the rationale for cleaning of medical equipment or devices before any further processing and includes examples of infectious complications associated with inadequate cleaning. In addition, discussion is presented concerning the relationship between cleaning and any subsequent processing before patient use. Guidance relative to the use of various cleaning agents is also provided. Additional emphasis on the necessity of following manufacturer's recommendations on cleaning agents and procedures is noted. PMID- 11858423 TI - Nosocomial infection among immunosuppressed patients in the intensive care unit. AB - Advances in technology and development of multidrug-resistant antibiotics have increased the number of immunosuppressed patients in need of crisis care and the incidence of nosocomial infection in the nation's hospitals. Nosocomial infections are proving particularly difficult to prevent in intensive care units (ICUs), where special care must be taken to avoid transmission of any infection. The authors present an overview of this problem in the ICU, outlining basic measures to be taken in preventing nosocomial infections and ensuring the safety of the patients. PMID- 11858424 TI - Infected wound management: advanced technologies, moisture-retentive dressings, and die-hard methods. AB - Wound infection is a significant problem for the complicated, critically ill patient. A critical care patient's plan of care can be challenging enough without complicating it with the additional comorbidity of a wound infection. Wound infection delays wound closure, disrupts wound tensile strength; increases hospital length of stay and costs; and escalates the patient's risk of bacteremia, sepsis, multisystem organ failure, and death. The goal is to reduce and eliminate the wound infection before it leads to such drastic consequences, especially in the age of antibiotic-resistant organisms. It is paramount to identify classic and not-so-obvious signs and symptoms of wound infections, correctly collect a wound specimen, and assist in appropriate systemic and topical wound management. Techniques to prevent wound infection and reduce bioburden include nontoxic wound cleansing, debridement of necrotic tissue, proper antibiotic management, and appropriate use of moisture-retentive dressings. Advanced technologies in moisture-retentive dressings include sustained-release silver and cadexomer iodine antimicrobial dressings and negative-pressure wound therapy. Accurate wound assessment, knowledge of new technologies, and applying current wound care standards to clinical practice will assist the critical care nurse in treating and preventing wound infections. PMID- 11858425 TI - Parent-child patterns of coping. AB - Coping of parents and children in a children's surgical area, where members had the same or different coping styles, was investigated using self-report and observation. Differences in effectiveness of coping strategies were found between groups of children, mainly in relation to emotion-focused coping. Parent groups were mainly distinguished from each other by the reported effectiveness of their spiritual and distracting strategies. Observational data showed group differences in patterns of parent-child interaction relating to coping behaviours prior to surgery. The findings indicate reciprocal parent-child influences of coping strategies and behaviours. PMID- 11858426 TI - Long-gap oesophageal atresia: a nursing perspective. AB - Oesophageal atresia (OA) is a congenital anomaly in which the oesophagus ends in a blind upper pouch. Specialised family-centred nursing care is required for the neonate/infant with 'long-gap' OA. Goals of preoperative nursing care focus on prevention of aspiration, optimising nutrition and thus growth of the baby and supporting parents. Primary repair is delayed to enable further growth of the infant and additional oesophageal growth. Additional nursing and medical research needs to be undertaken in a number of areas relating to the management of infants with long-gap OA. PMID- 11858427 TI - Is partnership in A&E feasible? AB - Partnership in care is frequently discussed within the discipline of children's nursing. There have been few definitions as to its meaning and therefore how it should be applied within the different specialties of children's nursing. This literature review identified a lack of research regarding partnership in care within the Accident and Emergency environment. PMID- 11858428 TI - Process recording: a student's perspective. AB - The framework of process recording is a useful tool to facilitate self-assessment and evaluation of interactions. Heron's (1986) Six Category Intervention Analysis Model enables the health professional to focus on the intention of their intervention. Health interviews in school frequently provide occasion for opportunistic health promotion, a large part of the school nurse's role. Health rarely motivates children, they perceive their vulnerability as low, but age, social class, and gender influence this. Children are often egocentric; inexperience limits a child's moral and cognitive judgements. PMID- 11858429 TI - Partnership in nursing care (PINC): the Blackburn model. AB - Concerns that partnership in care was not really working brought about the need to establish new documentation. A written management plan of care which actively involves the child and family to identify the partnership in care component is essential. Integration of medical and nursing history forms the basic assessment for a jointly managed plan of care. A major culture change needed to be managed to ensure more purposeful and effective communication within the nursing team. The balance of power for nursing care delivery has shifted away from the children's nurse towards the child/parent/carer. The Blackburn PINC model has provided the structures to ensure partnership in care is a reality. Ongoing development and evaluation in response to nursing and parent comments ensures that the process is responsive to current needs. PMID- 11858430 TI - Harry Potter and...the Editor's daydreams? PMID- 11858431 TI - FSID: the Foundation for the Study of Infant Deaths. PMID- 11858432 TI - Temperature taking in children. AB - Tempa-Dot single-use thermometers have been compared with mercury and tympanic thermometers in a paediatric setting. The largest difference in mean temperature measured between devices was less than 0.2 degrees C. Differences between devices are less than the random scatter in any individual measurement. It is argued that differences in measured temperature between thermometers are not clinically significant. Tempa-Dot thermometers can be used with confidence to measure the temperatures of young children PMID- 11858433 TI - Where are we heading in healthcare? PMID- 11858434 TI - Quick reference helps new nurses survive after orientation. PMID- 11858435 TI - Clarifying use of antipsychotic medications. PMID- 11858436 TI - Use infusion pumps to calculate dosages. PMID- 11858437 TI - Intravenous lidocaine questioned. PMID- 11858438 TI - Ethanol-induced therapeutic myocardial infarction to treat hypertrophic obstructive cardiomyopathy. PMID- 11858439 TI - Managing heart failure: a case study approach. AB - Heart failure is a common reason for admission to the hospital and to critical care units. The care of patients with heart failure is changing almost daily as new research and therapies become available. Nurses caring for these patients must use available information and assessment findings to discern which type of heart failure exists in each patient. In this way, the care provided can be enhanced, and outcomes can be optimized. Critically thinking nurses can positively influence patients' quality of life and potentially reduce the devastating morbidity and mortality associated with heart failure. PMID- 11858440 TI - Using continuous quality improvement techniques to determine the causes of hospital readmission. AB - This 6-month retrospective study provided our medical center with a profile of the population of patients readmitted within 31 days of discharge. We found that chronic illness and age greater than 65 years were the high risk factors related to patient readmissions. These 2 attributes are common in critical care patients. In addition, the study indicated that data input and collection procedures must be correctly followed in order to have accurate information on admission and readmission. Accurate information is important to management and external agencies. Accuracy is also important so that future problems with continuous quality improvement can be recognized and resources properly assigned. Recommendations for improving the data collection and reporting procedures and for improving the readmission rates were made to management, to the quality improvement coordinator, and to the quality improvement/risk management director. PMID- 11858441 TI - Evaluating healthcare information on the Internet: guidelines for nurses. PMID- 11858443 TI - Revisiting hypothermia: a critical concept. PMID- 11858444 TI - Clinical inquiry. PMID- 11858442 TI - Treating severe anemia in a trauma patient who is a Jehovah's witness. PMID- 11858445 TI - Is it acceptable to stop an infusion to obtain a blood sample? PMID- 11858446 TI - Evidence-based practice: a new name for an old concept? PMID- 11858447 TI - Exploration of diagnostic techniques for malignant melanoma: an integrative review. AB - The aim of this study is to explore the various diagnostic techniques for melanoma and assess their usefulness in the clinical practice of nurse practitioners. After a systematic review of the literature, 55 articles were closely reviewed, and from these, 26 studies were selected. These were original studies in English that examined diagnostic techniques that would improve accuracy for melanoma detection with a formal methods and results section. Six general categories dealing with diagnostic techniques for melanoma were found in the literature. These were naked-eye clinical examination alone, clinical examination with the aid of total-body photographs, epiluminescence microscopy (ELM), digital ELM, computer-assisted techniques, and teledermatology. Because of the research citing the poor diagnostic accuracy (DA) of nondermatologists, increased DA with dermatologists experienced in ELM techniques, and the importance of early melanoma diagnosis, the recommendation is to refer patients with suspicious pigmented skin lesions to experienced dermatologists, preferably those who use ELM or digital ELM. PMID- 11858448 TI - Dietary fiber and type 2 diabetes. AB - This article addresses the current theory, research, and implications of dietary fiber in the management of type 2 diabetes mellitus (DM; non-insulin-dependent DM). Dietary fiber shows promise in the management of type 2 DM. The inclusion of sufficient dietary fiber in a meal flattens the postprandial glycemic and insulinemic excursions and favorably influences plasma lipid levels in patients with type 2 DM. Water-soluble fiber appears to have a greater potential to reduce postprandial blood glucose, insulin, and serum lipid levels than insoluble fiber. Viscosity of the dietary fiber is important; the greater the viscosity, the greater the effect. Possible mechanisms for metabolic improvements with dietary fiber include delay of glucose absorption, increase in hepatic extraction of insulin, increased insulin sensitivity at the cellular level, and binding of bile acids. Patients with type 2 DM should increase their dietary fiber intake to 20 to 35 g/d and be aware of the considerations when increasing fiber intake. The nurse practitioner is in an ideal position to promote dietary fiber intake in such patients. PMID- 11858449 TI - Postmenopausal hormone replacement: historical perspectives and current concerns. AB - Despite continuing evidence of its value, hormone replacement (HR) therapy is used by only a minority of postmenopausal women. Some of this reluctance may stem from concerns about the medicalization of menopause and the labeling of menopause as a state of failure or disease that needs to be treated. These concerns are understandable given the impact of previous efforts to apply erroneous biological models to women's physiology, often to their detriment. Some may assert that current attempts to describe menopause as a state of estrogen deficiency are as wrong as previous explanations that the backing up of menstrual blood required purging and bleeding. However, there is an abundance of research attesting to the value of HR in decreasing such menopausal symptoms as hot flashes and insomnia and in preventing chronic problems, including urogenital atrophy and osteoporosis. In addition, recent research suggests that estrogen may have positive effects on cognition. Questions about HR and breast cancer remain, and several studies have found a small increase in breast cancer among long-term estrogen users. The recent introduction of selective estrogen response modifiers may further increase the safety of HR therapy. Many women worried about breast cancer and other possible estrogenic effects of HR are seeking approaches they consider more natural to managing menopause, turning to such untested remedies as soy supplements and herbs. While awaiting rigorous trials, clinicians can help patients understand the consequences of relying on therapies that have unknown long-term safety and effectiveness. PMID- 11858450 TI - A multimethod study of advanced practice nurse postdischarge care. AB - In the United States, increasing emphasis is being placed on understanding the roles and cost-effectiveness of advanced practice nurses (APNs). This study used both quantitative and qualitative strategies to explore the impact of APN intervention for frail rural elders being discharged from the hospital. For the quantitative component, analyses of specific postdischarge elder (n = 140) and caregiver (n = 65) outcomes were conducted using four groups receiving the following types of care: no nursing care, registered nurse (RN) care only, APN care only, and both RN and APN care. Outcomes assessed included elder cognitive functioning, self-rated health, informal services provided, and use of health care resources. Caregiver outcomes included physical, emotional, and depressive symptoms, as well as stress scores, physician visits, and missed work days. Elders in the APN-only group experienced fewer emergency room visits and hospital readmissions, although the difference was not significant. Caregivers receiving APN-only support reported significantly fewer work days missed. For the qualitative component, a focus group with four APNs who delivered the intervention was conducted to explore unique aspects of their roles and specific interventions they provided. A theme of these results was more comprehensive and autonomous delivery and management of care to both elders and their caregivers. PMID- 11858451 TI - Nurse/parent role perceptions in care of neonatal intensive care unit infants: implications for the advanced practice nurse. AB - This study compared parent and nurse perceptions of the nurse's roles regarding responsibilities toward infants in the neonatal intensive care unit (NICU). It also examined the attitudes of nurses and parents regarding the extent to which parents should participate in the care of their infants in the NICU, as well as the role of the advanced practice nurse (APN). The convenience sample of 25 parents of infants in the NICU and 35 nurses who cared for the infants was surveyed regarding perceptions of nurses and parents about nurse responsibilities and parent roles in the NICU. Results suggest parents and nurses have different perceptions about role expectations and that nurses perceive themselves to lack comfort and knowledge in providing support to parents. The findings support a role of the APN as fostering a nursing NICU philosophy to facilitate role transition for parents of infants in the NICU. PMID- 11858452 TI - Smoking patterns, health behaviors, and health-risk behaviors of college women. AB - In 1993, 22% of college women smoked; in 1997, the rate increased to 29%. College age women (<24 years) showed the greatest increase in smoking. The purpose of this study is to describe smoking behaviors of college women. The sample included 21 college-age female smokers. Each woman was interviewed about smoking habits and completed a health survey, the Fagerstrom Test for Nicotine Dependence, a Self-Efficacy Scale for predicting smoking relapse, a Decisional Balance Scale for smoking, a readiness for change tool, and the Derogatis Stress Profile. The majority of the students began smoking at the age of 14 years or younger and smoked fewer than 10 cigarettes per day. The biggest obstacles to quitting were being around other smokers and social activities involving alcohol. These students did not smoke when ill and were interested in quitting smoking. Smoking frequency correlated significantly with dependency and stage of change. Advanced practice nurses have a unique opportunity to identify these young smokers, educate them about smoking-cessation options, and offer specific strategies to help these women stop smoking. PMID- 11858453 TI - Decisions behind career choice for nurse practitioners: independent versus collaborative practice and motivational-needs behavior. AB - Nurse practitioners (NPs) can take an active role in defining and establishing their careers. Prepared as advanced practice nurses with specific assessment skills, primary care NPs have the opportunity to become independent or collaborative practitioners. This report examines the published work in the area of practice choice and motivational-needs behavior. Interviews with collaborative and independent primary care NPs were conducted. Against the framework of well established personality testing methods, open-ended interview questions were developed to elicit specific motivational-needs-based behavior characteristics. The motivational needs examined included the need for achievement, power, and affiliation. The interview findings were then synthesized using needs-based behavior theory. This new platform for role decision ultimately can prepare NPs to make informed career choices. PMID- 11858454 TI - Health care on the Internet: creating a safe environment. PMID- 11858455 TI - Kahn, J. A., Chiou, V., Allen, J. D., Goodman, E., Perlman, S. E., & Emans, S. J. (1999). Beliefs about Papanicolaou smears and compliance with Papanicolaou smear follow-up in adolescents. Archives of Pediatric and Adolescent Medicine, 153, 1046-1054. PMID- 11858456 TI - Yusuf, H. R., Coronado, V. G., Averhoff, F. A., Maes, E. F., Rodewald, L. E., Battaglia, M. P., & Mahoney, F. J. (1999). Progress in coverage with hepatitis B vaccine among US children, 1994-1997. American Journal of Public Health, 89 (11), 1684-1689. PMID- 11858457 TI - New applications for an old lignified element staining reagent. AB - The use is reported of Mirande's reagent in epifluorescence microscopy which permits a clear distinction between cellulosic and lignified tissues. Homogeneous Prespermatophytae and gymnosperm xylem appeared entirely green with Mirande's reagent under ultraviolet excitation, whereas heteroxyled angiosperm wood showed a mixed pink and blue-green colour. This coloration was due to the fluorescence of cellulose, since certain elements in dicotyledonous wood (parenchyma, fibres, xylem rays) are not entirely lignified. Monocotyledonous (Poaceae) lignin showed an intense blue fluorescence due to hydroxycinnamic acids bound to the cell wall. The method showed that lignification occurs first in the middle lamella, and later in the secondary wall of xylem cells. In addition, this staining technique proved useful in the study of lignin and suberin deposition in response to various stress factors. PMID- 11858458 TI - Alkaline fixation-resistant acid phosphatases in human tissues: histochemical evidence for a new type of acid phosphatase in endothelial, endometrial and neuronal sites. AB - The effect of pH during formalin fixation on acid phosphatases in human tissues was studied. Lysosomal-type acid phosphatase was sensitive to alkaline fixation, being completely inactive after fixation at pH 9.0. Prostatic and tartrate resistant osteoclastic/macrophagic types were alkaline fixation-resistant, as was an acid phosphatase localized in endothelium, endometrial stromal cells and intestinal nerves. The latter activity was further separable into fluoride- and tartrate-sensitive beta-glycerophosphatase and fluoride-sensitive, tartrate resistant alpha-naphthyl phosphatase. The activities appeared to represent either different, tightly associated enzymes or separate activity centres of a single enzyme. Alkaline fixation-resistant alpha-naphthyl phosphatase at endothelial, endometrial and neuronal sites was also well demonstrated in unfixed or neutral formalin-fixed sections as tartrate-resistant activity similar to classical tartrate-resistant acid phosphatase, but these phosphatases appear to be antigenically different. Alkaline fixation-resistant acid phosphatase showed a restricted tissue distribution both in endothelium (mainly in vessels of abdominal organs) and at neuronal sites (only in intestinal nerves). Alkaline fixation-resistant acid phosphatase appears to represent a previously unknown or uncharacterized enzyme activity whose chemical properties could not be classified as any previously known type of acid or other phosphatases. PMID- 11858459 TI - Ultrastructural immunolocalization of basic fibroblast growth factor to lipid bodies and secretory granules in human mast cells. AB - Isolated human lung mast cells were used to identify subcellular sites of basic fibroblast growth factor using a postembedding immunogold method. The factor was present in quantity in secretory granules and cytoplasmic lipid bodies. Cisterns of smooth endoplasmic reticulum and ribosome clusters, closely associated with lipid bodies, contained the factor as did the nuclear matrix. Factor-positive lipid bodies were adjacent to nuclear pores and often indented perinuclear cisternae. Altered secretory granules with reduced density, characteristic of secretion by piecemeal degranulation in mast cells, showed reduced gold label for basic fibroblast growth factor; small, electron-lucent (80-100 nm) transport vesicles near altered granules were labelled for the factor. Since these mature mast cells do not display extensive arrays of classical secretory organelles, such as rough endoplasmic reticulum and Golgi structures, these new subcellular localizations for basic fibroblast growth factor suggest several possible alternative release routes for a cytokine devoid of a signal sequence characteristic of regulated secretory proteins. PMID- 11858460 TI - Immunocytochemical location of endogenous cytokinins in buds of kiwifruit (Actinidia deliciosa) during the first hours of in vitro culture. AB - Post-embedding immunocytochemical techniques using peroxidase-antiperoxidase as markers for light microscopy or immunoglobulin G-gold for electron microscopy respectively were used for the localization of cytokinins [9-beta-D-ribofuranosil N6-(delta2-isopentenil) adenina ([9R]iP), 9-beta-D-ribofuranosyl-zeatin ([9R]Z) and 9-beta-D-ribofuranosyl-dihydrozeatin ([9R](diH)Z)] in kiwifruit (Actinidia deliciosa) meristematic cells of the second nodal segment. Immunolocation at the cellular level was carried out in cells from explants grown during 16 and 72 h in liquid medium. Subcellular immunolocalization was performed in cells from explants grown for 35 d on agar solidified-medium and for 30 min, 4 and 16 h in liquid medium with cellulose plugs as explant support. Taken as a whole, the results obtained for Actinidia deliciosa show that the studied cytokinins change their location during the culture period, although they can always be found to a greater or lesser extent in the nucleus and the cytoplasm. For instance, [9R]Z appears in the cytoplasm and in the nucleus during the first hours of culture and later is the only one that appears located mainly in nucleus. On the other hand, [9R](diH)Z changes from being predominantly located in the nucleus to practically appearing only in the cytoplasm at the end of the culture period. [9R]iP is principally found up to 4 h of culture in the cytoplasm, and at 16 h is evenly distributed in all the subcellular compartments except in the chloroplast. The existence of a large amount of cytokinins in the nucleus during the first hours of culture compared with the immunolabelling density at 35 d is probably due to the activation of cell cycle mechanisms leading to organogenic development at the beginning of culture. PMID- 11858461 TI - Immunocytochemical localization of serotonin in embryos, larvae and adults of the lancelet, Branchiostoma floridae. AB - Serotonin (5-hydroxytryptamine) is a biogenic amine distributed throughout the metazoans and has an old evolutionary history. It is involved as a developmental signal in the early morphogenesis of both invertebrates and vertebrates, whereas in adults it acts mainly as a neurotransmitter and gastrointestinal hormone. In vertebrates, serotonin regulates the morphogenesis of the central nervous system and the specification of serotonergic as well as dopaminergic neurons. The present study uses, as an experimental model, an invertebrate chordate, the lancelet Branchiostomafloridae, characterized by its remarkable homologies with vertebrates that allows the 'bauplan' of the probable ancestor of vertebrates to be outlined. In particular, the involvement of serotonin as a developmental signal in embryos and larvae, as well as a neurotransmitter and gastrointestinal hormone in adult specimens of Branchiostoma floridae, gives further support to a common origin of cephalocordates and vertebrates. PMID- 11858463 TI - Subcellular localization of epidermal growth factor in human parotid gland. AB - The intracellular distribution of epidermal growth factor was investigated in human parotid gland by immunogold cytochemistry at the electron-microscopy level. Epidermal growth factor immunoreactivity was demonstrated in both acini and ducts. In acinar cells, secretory granules appeared moderately stained, clearly indicating that parotid gland contributes to salivary epidermal growth factor through granule exocytosis. In ductal cells, gold particles were found to decorate numerous cytoplasmic vesicles, particularly abundant in striated duct cells. Since epidermal growth factor reactive vesicles were seen not only at the cellular apex, but nearby lateral plasma membranes as well, it leads to the hypothesis that epidermal growth factor may be discharged both apically into the saliva, and basally into the interstitium. PMID- 11858462 TI - Alteration of the Bcl-2:Bax ratio in the placenta as pregnancy proceeds. AB - The placenta is the primary site of nutrient and gas exchange between mother and foetus. During human placental development, proliferation, differentiation and apoptosis occur at different stages. In order to clarify some of the molecular mechanisms underlying these events, we investigated the pattern of expression of two members of the Bcl-2 family in human placenta samples and compared them to the level of apoptosis detected by the TUNEL method. In particular, we evaluated the expression of Bcl-2 and Bax and their ratio during the first and third trimester. We found that Bcl-2 was generally expressed at low levels during the entire gestational period. On the other hand, Bax was low during the first trimester but increased towards the end of gestation. In accordance with the change of ratio of these two molecules, the increase of apoptotic cells was observable in the third trimester. These data indicate that Bcl-2 and Bax are spatio-temporally regulated during placental development and that the different expression of the above mentioned genes is at least in part responsible for the delicate balance between cell proliferation and programmed cell death in the human placenta during pregnancy. PMID- 11858464 TI - A cytochemical method for visualising blood vessels selectively in semithin LR white sections. AB - Traditional cytochemical techniques for illustrating blood vessels are usually unsuitable for use in high resolution, 0.35 microm-thick LR White sections due to the thinness of the tissue and, if collagenous, high background staining. We report that a modification of the periodic acid-thiocarbohydrazide-silver proteinate-silver enhancement method (PATCH-SP-SE) selectively visualises, with low background, even the smallest blood vessels in semithin LR White sections of highly collagenous tissues such as human parietal peritoneum. PMID- 11858465 TI - Complementary therapies and the M25. PMID- 11858466 TI - Tough places to be tender: contracting for happy or 'good enough' endings in therapeutic massage/bodywork? AB - Listening to others has inspired this paper, as they share their experience and analyze their practice, and in the process uncover what is quintessential to therapeutic work. Of course, stories and insights can illuminate our own reflections and in turn deepen our understanding of what it means to be a nurse and massage therapist. They can also challenge what we think to be truths and sacred cows. This paper is about how confusion can be our best teacher when we seek to understand notions of contact and engagement in therapeutic work. When a patient says 'yes' to massage/bodywork what do they expect? Perhaps the massage will ease backache or leave them feeling profoundly relaxed; or could it be that the human contact is the most important need, but that need may be harder to express or even acknowledge? In examining the issues raised by this paper the author proposes a 'structural, emotional and energetic' (SEE) model for massage/bodywork contracts, and recommends that therapists seek supervision and support for their work. PMID- 11858467 TI - Complementary therapy programme at St Luke's Hospice, Plymouth. PMID- 11858468 TI - Should nurses practise complementary therapies? AB - This paper is based on the review of the literature that was undertaken as part of a Research Dissertation, submitted as part of an MSc in Nursing at Coventry University. The subsequent study was a comparison of Nurse and Non-nurse Aromatherapists, exploring their beliefs about the therapy and their reasons for practice. The author questions whether nurses should practise complementary therapies and if so why? If the nursing curriculum is to be developed to incorporate complementary therapies then we must be able to justify their inclusion in terms of patient outcomes, rather than in terms of meeting the needs of the nurse. The paper raises many questions, not all of which can be answered at this stage, but which can be considered and reflected on in depth. PMID- 11858469 TI - An interview with Kate Neil: nutritional therapy explained. Interview by Denise Rankin-Box. AB - Kate Neil is a Visiting Lecturer at the University of Westminster for their BSc Health Sciences: Nutritional Therapy degree course. Kate was part of the team that established this course 2 years ago, which is the first of its kind in this country and supervised their clinical training programme. Previously, Kate was the Director of Education Studies and Principal of the Institute for Optimum Nutrition in London. She directs her own company NS3UK which offers nutrition services to health care professionals (including a 3-year nutrition course in Dorking), industry and the public. Kate frequently contributes to professional and public journals and is Editor of the professional journal The Nutrition Practitioner. She is also author of Balancing Hormones Naturally. She practises from Harley Street and from NS3UK in Berkshire. She can be contacted at NS3 UK. Tel.: 01344 360033; E-mail: kate@ns3.co.uk and www.ns3.co.uk. Kate is currently studying for an MSc in Nutritional Medicine at Surrey University. PMID- 11858470 TI - Meeting the needs of people with cancer for support and self-management. AB - Health professionals tend to consider that they are in the best position to decide treatment options for cancer patients despite research evidence that a feeling of powerlessness can affect health outcomes and the inclusion of patient empowerment within government policy. This article describes a focus group study, carried out in collaboration with the University of Warwick, aiming to elicit the needs of people with cancer and their carers/supporters, the reactions to these needs and the strategies adopted to obtain support and self-management skills. The methodology was II two-stage focus groups carried out in four English cities in 1998, involving 54 people with cancer and 14 supporters. The main study findings identified a parallel 'journey' to the previously described emotional and medical cancer journey in the form of an epistemological journey of 'identity shifts' as people progressed through their disease. People required additional emotional and psychological support when these shifts were taking place and used a variety of strategies to obtain this, including the use of complementary therapies and other strategies designed to increase their sense of control. The article concludes by describing a 'holistic' approach to cancer care which supports patient empowerment and recommendations for implementing the research findings into practice. PMID- 11858471 TI - Migraine and photic stimulation: report on a survey of migraineurs using flickering light therapy. AB - Users of a commercial light therapy device who were using it to treat migraine were surveyed prospectively to determine what results they obtained after 30 days of daily use. Out of a total of 55 migraineurs, 44% reported that the frequency of their migraine attacks after the treatment was 'Somewhat Less' or 'Much Less' (under a conservative interpretation of these categories). Considering only the 28 migraineurs who stated that their migraine attacks were normally preceded by warning signs, 53% reported that the frequency of their migraine attacks was 'Somewhat Less' or 'Much Less'. In view of the limited efficacy and undesirable side-effects of the available migraine preventive drugs, photic stimulation (flickering light therapy) must be considered a possible preventive treatment for migraine. PMID- 11858472 TI - Management of pain through autogenic training. AB - Physical and emotional pain are an inevitable part of human existence and are without natural antidotes. In view of this, and in the light of increasing professional reluctance to depend on analgesics, this paper proposes the widespread application of autogenic training, a relaxation technique which has been seen to confront pain very effectively, and also to reduce substantially drugs dependency. It analyses autogenic training in respect of some of the more common pain-allied disorders such as childbirth, headaches and migraines, back pain, cancer and palliative care, and cardiology. PMID- 11858473 TI - 'Naturalistic vs reductionistic approaches to health-related practice: opposing dichotomy or symbiotic partnership?'. AB - Complementary therapies, within Health Service arenas, have traditionally been associated with 'naturalistic' approaches to health care provision rather than with 'reductionist' practices. Evidence does exist, however, that certain approaches to complementary therapies can exist comfortably within both camps. Subsequent debates within nursing literature, surrounding the place and validity of reductionist approaches to health care provision and their relationship with the 'counter-part' naturalistic (i.e. empowerment) approaches to health care, have existed for some time now. Naturalistic (inductive and interpretive) and reductionistic (deductive and fixed) classifications of health care provision have continued to be viewed, by many health care professionals, as apposite, divided and allopathic. This appears to be even more so recently where elements of reductionist health care have been portrayed in terms that serve to undervalue and undermine its contribution. This is whilst naturalistic approaches, in far more favourable terms, have gone on to be 'championed' by many health professions. This account sets out to investigate how this situation impacts upon the discipline of complementary therapies. It seeks to do so by defining the nature and purpose of these differing approaches - particularly within the boundaries of health promotion activities. It goes on to suggest that our current practices/viewpoints, related to these particular approaches, could be considered in themselves to be flawed, limiting and reductionist with a potential to unwittingly create a counterproductive practice ethic. As an alternative to this situation, it is suggested that by identifying the strengths and weaknesses of both stances it is possible to find common ground which marries together the more favourable aspects of these approaches. This can subsequently provide a clearer and more productive consensus for complementary therapies and other naturalistic based practices to move forward. PMID- 11858474 TI - Pregnancy, heparin and osteoporosis. PMID- 11858475 TI - Postpartum bone mineral density in women treated for thromboprophylaxis with unfractionated heparin or LMW heparin. AB - Venous thromboembolism remains an important cause of maternal mortality. In a randomised open study, 44 pregnant women with confirmed previous or current thromboembolism were randomised to receive either low-molecular-weight heparin, dalteparin (N = 21) once daily subcutaneously or unfractionated sodium heparin (UF heparin, N = 23) twice daily subcutaneously for thromboprophylaxis during pregnancy and puerperium. Bone mineral density (BMD) in the lumbosacral spine was measured with dual X-ray absorptiometry (DEXA) 1, 6, 16, 52 weeks and, if possible, 3 years after delivery. BMD values were also compared with those of healthy, delivered women (N = 19). Mean BMD of the lumbar spine was significantly lower in the unfractionated heparin group compared with the dalteparin and with the control groups (repeated measures ANOVA p = 0.02). BMD in the dalteparin group did not differ from BMD of healthy delivered women. Multiple logistic regression analysis revealed that therapy was the only independent factor influencing BMD at weeks 16 and 52. Therefore we recommend use of dalteparin instead of UF heparin for long-term thromboprophylaxis during and after pregnancy. PMID- 11858476 TI - The role of alpha(v)beta3 integrins in vascular healing. AB - Alpha(v)beta3 integrins play an important role in vascular healing. Vascular injury is a stimulus for expression of alpha(v)beta3 by vascular cells and, among other effects, alpha(v)beta3 integrins function in the adhesion of activated platelets to endothelium. white cell/endothelium interactions, platelet-mediated thrombin generation, fibrin clot retraction by nucleated cells, smooth muscle cell (SMC) migration and proliferation, vascular cell apoptosis, and vascular remodeling. There are ten different animal models in which treatment with alpha(v)beta3 antagonists reduced the vascular response, including (neo)intima formation, after mechanical injury. These studies, along with mechanistic data derived from cell culture studies, provide compelling evidence that alpha(v)beta3 integrins are involved in vascular repair processes. The challenge is to develop a therapeutic agent that will prove effective in reducing restenosis in humans following percutaneous coronary intervention (PCI). PMID- 11858477 TI - Association of factor V Leiden mutation with delayed graft function, acute rejection episodes and long-term graft dysfunction in kidney transplant recipients. AB - We analysed whether the factor V Leiden mutation--the most common hereditary predisposing factor for venous thrombosis--is associated with early and long-term graft dysfunction after kidney transplantation in 394 Caucasian kidney transplant recipients. The presence of factor V Leiden mutation was identified by allele specific PCR. The prevalence of the factor V Leiden mutation was compared to 32216 unselected neonates. The prevalence of the factor V Leiden mutation (GA genotype) was similar in 394 kidney transplant recipients and 32216 neonates. The frequency of known factors predicting long-term graft function were similar in patients with the GA genotype and with the normal factor V gene (GG genotype). The GA genotype was associated with the occurrence of no primary graft function (risk: 2.87: 95% confidence interval: 1.01-8.26; p < 0.05), the number of dialysis after transplantation in patients with no primary graft function until graft function (7.5 +/- 2.06 dialysis in GA patients; 4.2 +/- 0.36 dialyses in GG patients; p < 0.05), and the risk for at least one acute rejection episode (risk: 3.83; 95% confidence interval: 1.38-10.59; p < 0.02). The slope of l/creatinine per year was significantly lower in patients with the GA genotype (GA patients: 0.0204 +/- 0.008 dl/mg per year; GG patients: 0.0104 +/- 0.004 dl/mg per year; p < 0.02). The annual enhancement of the daily protein excretion rate was elevated in patients with the GA genotype (GA patients: 38.5 +/- 16.6 mg/24 h per year; GG patients: 4.9 +/- 4.4 mg/24 h per year; p < 0.02). Our study showed that the factor V Leiden mutation is associated with the occurrence of delayed graft function, acute rejection episodes and chronic graft dysfunction after kidney transplantation. PMID- 11858478 TI - Factor VII activity is an independent predictor of cardiovascular mortality in elderly women of a Sicilian population: results of an 11-year follow-up. AB - The aim of the Epidemiological project "Ventimiglia di Sicilia" is to identify the cardiovascular risk factors in a Sicilian population with a low risk profile and healthy nutritional habits. The risk of cardiovascular mortality in older subjects (over 60 years of age) is presented for an 11 year follow-up. Females showed higher prevalence of diabetes mellitus, hypertension, obesity and higher levels of total, LDL and HDL cholesterol, factor VII activity and fibrinogen compared to males. Cardiovascular mortality was related to hypertension and obesity in males, to high factor VII activity, obesity and diabetes mellitus in females. In a Logistic Regression model the same variables were independently correlated to cardiovascular mortality with the exception of obesity. In conclusion, these findings suggest that in a population with a low risk profile, other factors, such as factor VII activity, may emerge as predictors of cardiovascular mortality. PMID- 11858479 TI - Prospective study on soluble thrombomodulin and von Willebrand factor and the risk of ischemic and hemorrhagic stroke. AB - The aim of the present study was to examine if soluble thrombomodulin (sTM) and von Willebrand factor (VWF) could predict a first-ever ischemic or hemorrhagic stroke. This study was an incident case-referent study from within a population based cohort in northern Sweden. Up to 1996 about 44,000 subjects had been screened and stroke cases were classified according to the WHO MONICA criteria. A first-ever stroke occurred in 108 cases. A total of 216 controls were selected from the same cohort. This prospective study found no association with sTM or VWF and the development of a first-ever ischemic stroke (n = 87) in the logistic regression model. For the hemorrhagic stroke cases (n = 18), the multivariate logistic regression model revealed a significant negative association with sTM. When dichotomized, the upper level (>17.3 microg/L) of sTM, as compared with the lower level (<17.3 microg/L), showed one fifth of the risk for hemorrhagic stroke (OR, 0.18; CI, 0.05 to 0.69). No significant association was found for VWF. We suggest that the novel finding of an inverse relation between sTM and hemorrhagic stroke should be investigated in a larger study. PMID- 11858481 TI - Efficacy and safety of the factor VIII/von Willebrand factor concentrate, haemate P/humate-P: ristocetin cofactor unit dosing in patients with von Willebrand disease. AB - The present study was initiated to evaluate the safety and efficacy of Haemate-P (Humate-P in North America) (anti-hemophilic FVIII/VWF complex [human] dried, pasteurized) dosed in ristocetin cofactor units (VWF:RCo) in the treatment of von Willebrand disease (VWD) patients in Canada. This retrospective data collection reviewed the medical records of VWD patients treated under the Canadian Emergency Drug Release Program from November 22, 1991, to April 30, 1996. Data collection was accomplished by on-site retrieval from source data for 97 patients. Dosing was based on the German package insert, which lists only Factor VIII:C (FVIII:C) units, which were converted in the study analysis to VWF:RCo units based on the analysis of the individual manufactured lots of product used in these patients (average ratio of 2.6 IU VWF:RCo per IU FVIII:C). Twenty five different lots of Haemate-P/Humate-P were used to treat 437 different events in the 97 study patients (344 hemorrhagic events, 73 surgical interventions and 20 prophylactic infusion cycles). Overall, the median dose of concentrate per infusion used to treat surgical events was 69.1 IU VWF:RCo/kg (range 11.9-222.8); bleeding events 55.3 IU VWF:RCo/kg (range 17.1-227.5) and prophylaxis 41.6 IU VWF:RCo/kg (range 34.6-81.0). Treatment periods varied, with the majority of events treated for < or = 10 days (91%). Fifty percent of events that were treated longer than 10 days were given for prophylactic reasons. Efficacy was determined in a standardized manner by the physician, based on dosing in VWF:RCo activity. An overall clinical result of "excellent" or "good" was reported in 97% (424/437) of treatment events. A pediatric sub-population analysis of the patient population reported "excellent/good" efficacy in 100% (17/17) of treatment events in infants, 95% (155/164) in children, and 94% (76/81) in adolescent patients. Related adverse events (AEs) were observed in only 4 (4%) patients and were not deemed to be serious. The findings in this study confirm the safety and efficacy of Haemate P/Humate-P using VWF:RCo dosing in pediatric and adult patients with various types of VWD. PMID- 11858480 TI - The tissue factor and plasminogen activator inhibitor type-1 response in pediatric sepsis-induced multiple organ failure. AB - STUDY OBJECTIVE: Cytokines increase endothelial tissue factor (TF) and tissue plasminogen activator inhibitor type-1 (PAI-1) expression in vitro. Tissue factor interacts with factor VII to facilitate thrombosis and PAI-1 inhibits fibrinolysis by endogenous plasminogen activators. Because cytokine release is increased in children with sepsis-induced multiple organ failure (MOF), we hypothesized a cytokine associated increase in circulating TF and PAI-1 antigen release, and systemic activity in these patients. STUDY DESIGN: One hundred and seven consecutive children, who met the criteria for sepsis, and 10 critically ill children without sepsis, were enrolled in the study. Plasma TF and PAI-1 antigen and activity levels, Interleukin-6 antigen levels (IL-6), nitrite + nitrate levels (marker of nitric oxide production) and number of organs failing were measured on days 1-3 of sepsis. RESULTS: Increased TF and PAI-1 antigen, and PAI-1 activity levels were associated with increasing IL-6 and nitrite + nitrate levels (p <0.05), the development of MOF (p <0.05), and mortality (p <0.05). Increased systemic PAI-1 activity was associated with cardiovascular, renal. and hepatic failure (p <0.05). Increased systemic TF activity was associated with the development of coagulopathy (p <0.05) and tended to be associated with mortality (p = 0.06, power .77) CONCLUSIONS: A shift to an anti-fibrinolytic endothelium phenotype characterizes children who develop sepsis-induced MOF and mortality. Children with coagulopathy have a shift to a pro-coagulant phenotype. These findings support potential therapeutic roles for PAI-1 and TF pathway inhibitors in reversal of this devastating pathophysiologic process. PMID- 11858482 TI - A dose-ranging study of the oral direct thrombin inhibitor, ximelagatran, and its subcutaneous form, melagatran, compared with dalteparin in the prophylaxis of thromboembolism after hip or knee replacement: METHRO I. MElagatran for THRombin inhibition in Orthopaedic surgery. AB - The novel, oral direct thrombin inhibitor, ximelagatran (formerly H 376/95), represents an advance in antithrombotic therapy through its oral availability. After oral administration, ximelagatran is converted to its active form, melagatran. Melagatran can also be administered subcutaneously (s.c.). The results from the first clinical study with ximelagatran are reported. In this randomized, parallel-group, controlled study, 103 patients scheduled for elective total hip or total knee replacement received s.c. melagatran (1, 2 or 4 mg bid) for 2 days commencing immediately before surgery, followed by oral ximelagatran (6, 12 or 24 mg bid) for 6-9 days. Another 33 patients received dalteparin 5000 IU s.c. once daily, started the evening before surgery, for 8-11 days. At bilateral venography, deep vein thrombosis was found in 20.5% (16/78) of patients who had received s.c. melagatran and oral ximelagatran and in 18.5% (5/27) of patients in the dalteparin group. The study did not evaluate a dose-response for efficacy, and no differences between the three dose levels of melagatran and ximelagatran were shown. No pulmonary embolism was diagnosed during treatment. Total bleeding in the s.c. melagatran plus oral ximelagatran groups showed no dose-response and was similar to that seen in the dalteparin group. The pharmacokinetic properties of melagatran in the surgery patients were consistent with those observed for healthy subjects, and the APTT ratio, which increased non linearly with plasma melagatran concentration, showed a consistent concentration effect relationship during the treatment period. Ximelagatran and melagatran were well tolerated. In conclusion, ximelagatran and its active form melagatran appear to be promising agents for the prevention of venous thromboembolism following orthopaedic surgery. PMID- 11858483 TI - Effect of new synthetic heparin mimetics on whole blood thrombin generation in vivo and in vitro in rats. AB - The effect of new heparin mimetics (synthetic oligosaccharides) was studied in vitro with regard to thrombin generation (TG) in rat platelet rich plasma (PRP) and whole blood (WB) and in vivo on stasis-induced venous thrombosis in the rat. TG in PRP and in WB was highly dependent on platelet count and strongly influenced by the haematocrit. The peak of TG appeared to be significantly higher in WB than in PRP whereas the endogenous thrombin potential (ETP) was not significantly different under either condition. The effect of hirudin, the synthetic pentasaccharide SR90107/Org31540 (SP) and heparin were measured on TG in PRP and WB. We then compared the effect of two new synthetic heparin mimetics (SR121903A and SanOrg123781) with potent and comparable antithrombin (AT) mediated activity against factor Xa and thrombin. These two compounds were made of a pentasaccharide with a high affinity to AT, prolonged at the non-reducing end by an oligosaccharide chain recognised by thrombin. In SR121903A, the charge density and charge distribution was analogous to that of heparin whereas in SanOrg123781 the charges were only located on the last 5 saccharides of the non reducing end of the molecule. In PRP and in WB, SR121903A acted on the lag time and on the AUC whereas SanOrg123781 inhibited thrombin formation with no effect on the lag time. SanOrg123781 was more potent in inhibiting TG than SR121903A. This difference was due to the structures of the compounds that differed in their ability to be neutralised by platelet factor 4. The antithrombotic effect of the two compounds was examined in a venous thrombosis model in rats. We observed that SanOrg123781 was more active than SR121903A and heparin. Taken together, these results indicate that the activity of oligosaccharides is greatly influenced by the global charge density of the molecule and show that SanOrg123781 is a potent and promising antithrombotic drug candidate. PMID- 11858484 TI - Topically administered macromolecular heparin proteoglycans inhibit thrombus growth in microvascular anastomoses. AB - Previously, during blood perfusion over collagen-coated surfaces; soluble or immobilized heparin proteoglycans (HEP-PG) have been shown to block thrombus growth. Our aim was to study the antithrombotic effect of locally applied unfractionated heparin (UFH, 1 mg/ml), or rat mast cell-derived HEP-PG (MW 750 kD, 10 microg/ml) compared with saline in early (10 min) and late (3 days) thrombus formation upon anastomosis of rat common femoral arteries. In both semiquantitative scanning electron microscopy (SEM) and quantitative platelet Indium 111-labeling HEP-PG inhibited thrombus growth in comparison with saline. At 10 min, the extent of thrombosis (scale 1-4) in SEM followed the order: saline (3.2+/-0.8) > UFH (2.8+/-1.0) > HEP-PG (1.8+/-0.8), and also Indium 111-positive platelets (10(6)) accumulated on the anastomosed vessel in the same order 14.2 +/ 7.2, 10.3 +/-5.0, and 7.7 +/-3.1 (saline vs. HEP-PG, p = 0.03 and 0.05, respectively). At 3 days all HEP-PG-treated vessels remained patent with only small mural thrombi, whereas 2/7 saline- and 1/7 UFH-treated anastomoses occluded and showed more thrombosis overall. We conclude that locally administered HEP-PG inhibit arterial thrombus growth in anastomosed small-sized arteries and could prevent thrombotic complications in (micro)vascular surgery and arteriovenous shunts. PMID- 11858485 TI - Genetic and phenotypic variability between families with hereditary protein S deficiency. AB - While many mutations thought to result in protein S (PS) deficiency are known, there have been few attempts to relate genotype expression with plasma phenotype. We have investigated the nature and consequence of PS gene (PROS1) mutations in 17 PS-deficient families who presented with mixed type I and type III phenotypes. Seven different mutations were found in nine families: delG-34 (STOP codon at 24), Val-24Glu, Arg49Cys, Asn217Ser, Gly295Val, +5 G to A intron j and His623Pro. PS wild type (PSWT) and the five missense mutants were transiently expressed in COS-1 cells. All mutants expressed lower (p<0.05) PS antigen compared to PSWT (100%). The mutants Val-24Glu, Gly295Val and His623Pro expressed very low/undetectable PS levels. The mutant Asn217Ser produced around 30% of PSWT, while the mutant Arg49Cys had the highest PS levels (around 50%). Metabolic labelling and pulse-chase experiments showed that all of the mutants had impaired secretion, but this was of variable severity. Also, enhanced intracellular degradation of unsecreted material was found for all mutants. There was a strong correspondence between plasma free PS levels in carriers of the mutations, secreted PS from transfected COS-1 cells and labelled PS from 24 h conditioned medium in pulse-chase experiment. We conclude that the magnitude of secretion defect depends on the nature of the PROS1 mutation and influences the level of free PS in plasma. It is likely that the severity of the secretion defect will determine the risk for venous thrombosis. PMID- 11858486 TI - Single cell analysis of factor VIII-specific T cells in hemophilic mice after treatment with human factor VIII. AB - A multi-parameter flow-cytometry assay was established suitable for analyzing T cell-specific cell surface markers (CD3, CD4) together with intracellular cytokines on a single cell level. This assay was used to identify the frequency and the kinetic of different populations of factor VIII (FVIII)-specific CD4+ T cells in hemophilic E-17 mice after treatment with human FVIII. A clear temporal correlation was found between the appearance of FVIII-specific CD4+ T cells in the spleen and the detection of anti-FVIII antibodies in plasma. These cells and antibodies were detectable in all experiments after two doses of FVIII and in a few even after a single dose. The IFN-gamma-producing T cells were the most prominent type of FVIII-specific T cells suggesting Th1-type T cells have an important role in regulating the anti-FVIII immune response in E-17 mice. IL-10 producing T cells were the second most dominant type. They were detectable after two doses of FVIII and increased in frequency after four. Cytokine co-expression studies analyzing IL-10 and IFN-gamma in the same cell indicated that there might be at least two types of IL-10 positive T cells, those cells that produce IL-10 only and in addition cells that produce IL-10 and IFN-gamma. Furthermore, FVIII specific T cells producing IL-2 were found in all experiments after two doses of FVIII. In a few experiments IL-4-producing T cells were seen but in most experiments they were not detectable. In contrast, IL-4 could be found in supernatants of in vitro restimulated CD8- spleen cells. PMID- 11858487 TI - Non-inversion factor VIII mutations in 80 hemophilia A families including 24 with alloimmune responses. AB - Heteroduplex screening identified 74 small mutations in the factor VIII genes of 72 families with hemophilia A. In addition, patients from 3 families with high titer inhibitors had partial gene deletions and 5 unrelated families that were negative for heteroduplex formation had a mutation on direct sequencing. The latter had mild hemophilia A with an inhibitor, and sequencing their exon 23 fragments found a transition predicting a recurrent Arg2150 to His. Of 69 distinct mutations (including the 3 partial gene deletions), 47 are novel. Of small mutations, 51 were missense (one possibly a normal variant and two that could also alter splicing) at 39 sites, 13 were small deletions or insertions (3 inframe and one a normal variant in an intron), 13 were nonsense at 12 sites and 2 altered intron splice junctions. In 24 families, at least one affected member had evidence for an alloimmune response to factor VIII: of these, 11 were associated with missense mutations. In 14 families, de novo origin was demonstrated. PMID- 11858488 TI - Substitution of Gly-548 to Ala in the substrate binding pocket of prothrombin Perija leads to the loss of thrombin proteolytic activity. AB - Prothrombin Perija is a dysprothrombin derived from a homozygous patient that manifests low thrombin activity upon activation in a one-stage assay. Purified prothrombin Perija showed normal appearance on SDS-PAGE. and was cleaved normally to form alpha-thrombin by the prothrombinase complex. The activated form, thrombin Perija, however, did not show any proteolytic activity towards native substrates, fibrinogen, protein C or various synthetic substrates for alpha thrombin, but it was able to bind to antithrombin III, although the binding capacity was markedly reduced even in the presence of heparin. Thrombin Perija showed full reactivity toward a small inhibitor, DFP, indicating that the molecular defect is in the substrate binding site in the thrombin molecule but not in the active site itself. By DNA sequence analysis of the patient prothrombin gene, we identified a G to C mutation at nucleotide 20016 in exon 14, which predicts a Gly-548 to Ala substitution in the prothrombin Perija molecule. The structural modeling of thrombin Perija suggests that Ala-548 is located close to the limb of the cavity wall of the substrate binding pocket, and that the methyl group blocks protrusion of the guanidino group of Arg into the cavity. This steric hindrance may well inhibit the access of Arg-containing substrates to the catalytic Ser-525 leading to the loss of proteolytic activity. PMID- 11858489 TI - Bothrojaracin, a proexosite I ligand, inhibits factor Va-accelerated prothrombin activation. AB - Bothrojaracin (BJC) is a 27 kDa snake venom protein from Bothrops jararaca that has been characterized as a potent ligand (KD = 75 nM) of human prothrombin (Monteiro RQ, Bock PE, Bianconi ML, Zingali RB, Protein Sci 2001; 10: 1897-904). BJC binds to the partially exposed anion-binding exosite I (proexosite I) forming a stable 1:1, non-covalent complex with the zymogen whereas no interaction with fragment 1 or 2 domains is observed. In addition, BJC interacts with thrombin through exosites I and II (KD = 0.7 nM), and influences but does not block the proteinase catalytic site. In the present work we studied the effect of BJC on human prothrombin activation by factor Xa in the absence or in the presence of its cofactors, factor Va and phospholipids. In the absence of phospholipids, BJC strongly inhibited (80%) the zymogen activation by factor Xa in the presence but not in the absence of factor Va, suggesting a specific interference in the cofactor activity. In the presence of phospholipid vesicles (75% phosphatidylcholine, 25% phosphatidylserine), BJC also inhibited (35%) prothrombin activation by factor Xa in the presence but not in the absence of factor Va. BJC showed a higher inhibitory effect (70%) towards thrombin formation by prothrombinase complex assembled on phospholipid vesicles composed by 95% phosphatidylcholine, 5% phosphatidylserine. Activation of prothrombin by platelet assembled prothrombinase complex (factor Xa, factor Va and thrombin-activated platelets) showed that hirudin (SO3-) and BJC efficiently inhibit the thrombin formation (43% and 84%, respectively). Taken together, our results suggest that proexosite I blockage decreases the productive recognition of prothrombin as substrate by factor Xa-factor Va complex and prothrombinase complex. Furthermore, data obtained with human platelets suggest that proexosite I may play an important role in the physiological activation of prothrombin. PMID- 11858490 TI - Novel factor V C2-domain mutation (R2074H) in two families with factor V deficiency and bleeding. AB - The molecular basis of Factor V deficiency has been defined in few patients only. We report a homozygous nucleotide change (G6395A) in two Tunisian probands with Factor V deficiency and bleeding episodes. This substitution results in the replacement of an arginine (R) by a histidine (H) in amino acid position 2074, located in the Factor V C2-domain. Mutations in this protein domain have not previously been described. Several lines of evidence support that this sequence variant is indeed disease causing: 1) Crystal structures of Factor V and molecular C2-domain modeling studies of H2074 suggest that the conserved R2074 is required for correct folding; 2) Structure-function studies of selective Factor V mutants (R2074A) demonstrate the importance of R2074 for structural stability of the Factor V C2-domain and for cofactor activity (1); 3) In Factor VIII, point mutations in codon 2209, which corresponds to position 2074 in Factor V, cause hemophilia A. PMID- 11858491 TI - Inhibition of thrombin generation by the oral direct thrombin inhibitor ximelagatran in shed blood from healthy male subjects. AB - Ximelagatran, an oral direct thrombin inhibitor, whose active form is melagatran, was studied using a model of thrombin generation in humans. Healthy male volunteers (18 per group) received ximelagatran (60 mg p.o.), dalteparin (120 IU/kg s.c.) or a control (water p.o.). Shed blood, collected after incision of the forearm with standardised bleeding time devices at pre-dose, and at 2, 4 and 10 h post-dosing, was analysed for markers of thrombin generation. Statistically significant reductions (p < 0.05) in levels of prothrombin fragment 1+2 (F1+2) and thrombin-antithrombin complex (TAT) in shed blood were detected at 2 and 4 h post-dosing in both the ximelagatran and dalteparin groups. Shed blood F1+2 and TAT levels had returned to pre-dose levels at 10 h post-dosing. Using a shed blood model, we demonstrate that the reversible thrombin inhibitor melagatran and, therefore, oral administration of ximelagatran, inhibits thrombin generation in humans after acute activation of coagulation. PMID- 11858492 TI - Influence of t-pA and u-PA on adipose tissue development in a murine model of diet-induced obesity. AB - To investigate the potential role of tissue-type plasminogen activator (t-PA) or urokinase-type plasminogen activator (u-PA) in development of adipose tissue, we have used a nutritionally induced obesity model in t-PA (t-PA-/-) and u-PA (u-PA /-) deficient mice. Five week old male wild-type (WT), t-PA-/- or u-PA-/- mice (n = 9 to 16) were fed a high fat diet (HFD, 42% fat). After 16 weeks of HFD, the body weight of t-PA-/- mice was significantly higher than that of WT mice (48 +/- 1.1 g vs. 39 +/- 2.2 g, p = 0.004). The total weight of the isolated subcutaneous (sc) fat deposit was higher in t-PA-/- than in WT mice (2.4 +/- 0.22 g vs. 1.2 +/ 0.29 g, p = 0.002). accompanied with higher adipocyte diameters (80 +/- 1.7 microm vs. 61 +/- 4.7 microm, p < 0.01). These differences were not observed in the intra-abdominal fat deposit. The number of stroma cells in both adipose tissue territories was increased in t-PA-/- as compared to WT mice (2.0 +/- 0.13 vs. 1.5 +/- 0.10, p = 0.2 and 3.0 +/- 0.17 vs 1.6 +/- 0.17, p = 0.0001, stroma cells/adipocytes in sc and intra-abdominal tissue, respectively), partly as a result of an increased number of endothelial cells (192 +/- 9 vs. 154 +/-18, p = 0.06 and 108 +/- 13 vs. 69 +/- 8, p = 0.04 CD31 stained/adipocyte area). In contrast the weight gain and adipose tissue development in u-PA-/- mice was not different from that in WT mice. These data suggest that t-PA but not u-PA plays a role in adipose tissue development. PMID- 11858493 TI - Paradoxical platelet activation was not observed on dissociation of abciximab from GpIIb-IIIa complexes. AB - The ability of abciximab to bind and dissociate from platelets raises the question of the conformational state of GPIIb-IIIa complexes losing abciximab and the risk of paradoxical drug-induced platelet activation. Platelets incubated with abciximab and mixed in vitro with c7E3 Fab-free platelets lost the drug to the new platelets giving a single platelet population with a unimodal abciximab distribution within 17 h. Prelabeling the receiving platelets with phycoerythrin labeled anti-GPIb monoclonal antibody (MoAb), permitted their identification by flow cytometry. Binding of PAC-1 and AP6, two MoAbs specific for activated GPIIb IIIa, was then assessed to both losing and receiving platelet populations during transfer of abciximab. The subpopulation losing c7E3 Fab failed to show increased binding of these MoAbs. However, PAC-1 binding increased in both subpopulations after addition of ADP. Thus GPIIb-IIIa complexes are not in an activated state after dissociation of abciximab unless there is an additional source of activation. PMID- 11858494 TI - Ticlopidine inhibits the prothrombotic effects of thrombopoietin and ameliorates survival after supralethal total body irradiation. AB - Thrombopoietin modulates the response of platelets to several agonists and, on the other hand, those agonists can be released following irradiation. Thus, we have determined the effects of thrombopoietin, on its own and in combination with ticlopidine, an anti-platelet drug, on platelet activation, thrombosis formation and survival of irradiated C57BL6/J mice. Administration of thrombopoietin 2 h after 9 Gy total body irradiation increased the 125I-fibrin deposition in mouse tissues and accelerated platelet consumption as revealed by an enhanced drop in platelet counts. Additionally, the number of activated platelets, i.e. those expressing P-selectin on their membrane, was higher in thrombopoietin-treated mice as compared to the placebo group, regardless ex vivo stimulation with agonists. These effects of thrombopoietin on platelet activation and consumption were reduced when mice were pretreated with ticlopidine. The combination of ticlopidine with thrombopoietin almost fully promoted 180-day survival, reaching the same efficacy as bone marrow transplantation, while only 30% of the mice treated with thrombopoietin alone survived. In summary, thrombopoietin induces long term-mortality of irradiated mice probably through platelet-mediated thrombosis and thus, ticlopidine efficiently counteracts these adverse effects of thrombopoietin. PMID- 11858495 TI - Molecular modeling of the seven tandem leucine-rich repeats within the ligand binding region of platelet glycoprotein Ib alpha. AB - Platelet glycoprotein (GP)Ib-IX-V mediates von Willebrand Factor (vWF)-dependent adhesion to vascular subendothelium at high shear in (patho)physiological thrombus formation. The ligand-binding domain of GPIb-IX-V is within the N terminal 282 residues of GPIb alpha, that contains seven tandem leucine-rich repeats (Leu36-Ala200). Repeats 2-4 are critical for vWF binding. In this study, we have built molecular models of the seven leucine-rich repeats of human, canine and mouse GPIb alpha, providing novel insights into the species-specific interaction between human vWF and its receptor. Interestingly, a major difference between the models was a large negatively charged patch on the concave face of human, but not canine, repeats 2-4. In addition, five individual mutations within the leucine-rich repeats of GPIb alpha associated with the bleeding disorder Bernard-Soulier syndrome, that result in dysfunctional vWF binding, were mapped to the model of human GPIb alpha. This provides the basis for relating these genetic lesions to abnormal function of the receptor. PMID- 11858496 TI - A defect in collagen receptor-Ca2+ signaling system in platelets from cattle with Chediak-Higashi syndrome. AB - Decreased platelet aggregation to collagen is a cause for bleeding diathesis of Chediak-Higashi syndrome (CHS). We investigated whether the collagen receptor Ca2+ signaling system was impaired in platelets from cattle affected with CHS. A collagen-induced increase in cytosolic Ca2+ ([Ca2+]i) was depressed in CHS platelets, which was accompanied by a decrease in the production of inositol 1,4,5-trisphosphate. When the influences of endogenous arachidonic acid metabolites and ADP were excluded, convulxin or collagen-related peptide, which are specific agonists for the collagen receptor GPVI, increased [Ca2+]i in both normal and CHS platelets. In contrast, rhodocytin, which was thought to activate another collagen receptor GPIa/IIa, increased [Ca2+]i in CHS platelets to a lesser extent than in normal ones. Cytochalasin D, an actin polymerization inhibitor, depressed the response to collagen or rhodocytin but not the response to convulxin. Adhesion of CHS platelets to acid soluble type I collagen, which was mediated by GPIa/IIa, was similar to that of normal platelets. These results suggest that a defect in the rhodocytin-sensitive pathway is responsible for decreasing the response to collagen in CHS platelets. It remains to be determined which receptor is associated with the mechanism. PMID- 11858497 TI - Fatal pulmonary embolism immediately after transatlantic air travel to the United States: less than one in a million. PMID- 11858498 TI - Short-term exposure to high altitude causes coagulation activation and inhibits fibrinolysis. PMID- 11858499 TI - Platelet glycoprotein Ib and reactivity of platelets in diabetic and non-diabetic hemodialysed patients. PMID- 11858500 TI - Successful use of recombinant activated factor VII (Novoseven) in controlling severe intra-abdominal bleeding after liver needle biopsy. PMID- 11858501 TI - Endothelial nitric oxide gene polymorphism (Glu298Asp) is not associated with coronary artery disease in Turkish population. PMID- 11858502 TI - Genetic variation in coagulation factors II, V, VII and fatal MI. PMID- 11858503 TI - Weak regulation of protein Z biosynthesis by inflammatory cytokines. PMID- 11858504 TI - Single-dose pharmacokinetics of porcine factor VIII (Hyate C). PMID- 11858505 TI - Four novel polymorphisms in the fibrinogen Aalpha gene. PMID- 11858506 TI - Activated recombinant factor VII (rFVIIa) in bleeding management after therapy with IIb/IIIa-inhibitor tirofiban. PMID- 11858507 TI - Thrombosis-associated gene variants in sickle cell anemia. PMID- 11858508 TI - Recurrent late fetal death in women with and without thrombophilia. PMID- 11858509 TI - A rebuttal: Counseling pregnant women with factor V Leiden homozygosity: use absolute instead of relative risks. PMID- 11858510 TI - The implications of hormesis to ecotoxicology and ecological risk assessment. AB - Changes required for the explicit recognition of hormesis are outlined for both ecotoxicology and ecological risk assessment (ERA). A major research need is the extension of hormesis beyond chemical stressors to abiotic (e.g., habitat) and biotic stressors (e.g., species introductions, organism interactions). An overreaching research need is to determine for all stressors with model organisms, populations, and communities whether hormesis has positive, neutral, or adverse effects. The latter are the least likely; however, neutral effects cannot be ruled out. Based on our present state of knowledge, hormesis is likely to have more of an impact on ecotoxicology than on ERA. In the case of the latter, it is most likely to make a difference only in a detailed-level ecological risk assessment (DLERA), the most complex form of ERA. Further, for hormesis to be accepted fully into ecotoxicology or ERA will require a paradigm shift. Three ongoing paradigm shifts to which hormesis could be linked are: recognition of the low utility of no-observed-effects concentrations (NOECs); recognition of the need for special treatment of essential element dose concentration responses, which are similar to hormesis; and the replacement of environmental toxicology with ecological toxicology (ecotoxicology). PMID- 11858511 TI - Experiments, analyses and decisions: hormesis in ecotoxicology. AB - We have been challenged by Peter Chapman to consider the implication of hormesis for both ecotoxicology and ecological risk assessment. A consideration of his proposal led to the following observations and comments. I echo his comments related to the need for experimental designs and statistical tools that accommodate and address hormesis. I expand on his comments related to the basic question of how adverse responses should be defined for hormetic concentration response patterns for both individual species and populations. Finally, I reflect on the notion of the implication of incorrectly detecting or not detecting hormesis in an ecosystem. PMID- 11858512 TI - Commentary: role of hormesis in ecological risk assessment. PMID- 11858513 TI - The implications of hormesis to ecotoxicology and ecological risk assessment (ERA). PMID- 11858514 TI - Hormesis--does it have relevance at the population, community or ecosystem levels of organization? PMID- 11858515 TI - Hormesis in ecological risk assessment: a useful concept, a confusing term, and/or a distraction? PMID- 11858516 TI - Commentary: hormesis and ecological risk assessment. PMID- 11858517 TI - Focus on exposure-response relationships, and complex forms will come naturally. PMID- 11858518 TI - Pharmacokinetic and toxicology assessment of INTERCEPT (S-59 and UVA treated) platelets. AB - The pathogen inactivation process developed by Cerus and Baxter Healthcare Corporations uses the psoralen, S-59 (amotosalen) in an ex vivo photochemical treatment (PCT) process to inactivate viruses, bacteria, protozoans, and leukocytes in platelet concentrates and plasma. Studies were performed by intravenous infusion of S-59 PCT formulations +/- compound adsorption device (CAD) treatment and with non-UVA illuminated S-59, using doses that were multiples of potential clinical exposures. The studies comprised full pharmacokinetic, single- and repeated-dose (up to 13 weeks duration) toxicity, safety pharmacology (CNS, renal, and cardiovascular), reproductive toxicity, genotoxicity, carcinogenicity testing in the p53(+/-) mouse, vein irritation, and phototoxicity. No specific target organ toxicity (clinical or histopathological), reproductive toxicity, or carcinogenicity was observed. S-59 and/or PCT formulations demonstrated CNS, ECG, and phototoxicity only at supraclinical doses. Based on the extremely large safety margins (>30,000-fold expected clinical exposures), the CNS and ECG observations are not considered to have any toxicological relevance. Additionally, after a complete assessment, mutagenicity and phototoxicity results are not considered relevant for the proposed use of INTERCEPT platelets. Thus, the results of an extensive series of in vitro and in vivo studies have not demonstrated any toxicologically relevant effects of platelet concentrates prepared by the INTERCEPT system. PMID- 11858519 TI - Nanoparticle technology for drug delivery across the blood-brain barrier. AB - Nanoparticles (NP) are solid colloidal particles ranging in size from 1 to 1000 nm that are utilized as drug delivery agents. The use of NPs to deliver drugs to the brain across the blood-brain barrier (BBB) may provide a significant advantage to current strategies. The primary advantage of NP carrier technology is that NPs mask the blood-brain barrier limiting characteristics of the therapeutic drug molecule. Furthermore, this system may slow drug release in the brain, decreasing peripheral toxicity. This review evaluates previous strategies of brain drug delivery, discusses NP transport across the BBB, and describes primary methods of NP preparation and characterization. Further, influencing manufacturing factors (type of polymers and surfactants, NP size, and the drug molecule) are detailed in relation to movement of the drug delivery agent across the BBB. Currently, reports evaluating NPs for brain delivery have studied anesthetic and chemotherapeutic agents. These studies are reviewed for efficacy and mechanisms of transport. Physiological factors such as phagocytic activity of the reticuloendothelial system and protein opsonization may limit the amount of brain delivered drug and methods to avoid these issues are also discussed. NP technology appears to have significant promise in delivering therapeutic molecules across the BBB. PMID- 11858520 TI - Ophthalmic vehicles containing polymer-solubilized tropicamide: "in vitro/in vivo" evaluation. AB - Commercial 1.0% aqueous tropicamide (TR) eyedrops are buffered to pH 4.4-5.0 to produce sufficiently stable solutions of the weakly basic, poorly soluble drug. These acidic solutions, however, are irritants and may induce copious lachrimation, thus reducing the drug bioavailability. The aim of the present study was to evaluate some solubilizing agents for the preparation of 1.0% TR ophthalmic solutions adjusted at physiologically compatible pH, potentially showing increased eye tolerance, activity, and stability when compared with standard commercial eyedrops. The tested solubilizers were two non-ionic surfactants-Tyloxapol (TY) and Cremophor EL (CR) and one polymer, Pluronic P85 (PL). Four stable 1% TR formulations, containing 3% TY, 7.5% CR, 15% PL, or 5% CR + 10% PL were submitted to mydriatic activity tests in rabbits. They improved to a small but statistically significant extent the AUC for mydriatic effect of TR in the test animals when compared with commercial 1.0% TR eyedrops. PMID- 11858521 TI - Investigation of flow properties of powders by means of a uniaxial tester, in relation to direct tablet compression. AB - Unexpected poor flowability during commercial production of a direct compression tablet formulation initiated an investigation of the flow properties of the powder mixture and its components by means of a uniaxial tester. The failure function--a curve describing the strength of the powder bed as a junction of the maximum main stress that has consolidated the bed--of the powder mixture and its components was determined. The drug was more cohesive than the filler, which was somewhat more cohesive than the powder mixture. Three excipients--a hinder, a glidant and a lubricant constituting 3.5 w/w% of the formulation improved the flowability of the mixture of active ingredient and filler. The failure function discriminated powder mixtures with poor flow from mixtures with medium or good flow. However, it was not possible to discriminate medium from good flow by means of the failure function. Attempts to correlate univariately the flow property parameters of the powder mixtures with particle size data or flow property data of included active ingredient and filler batches failed. Therefore a multivariate approach was tested. Principal component analysis (PCA) and projection to latent structures by means of partial least squares (PLS) were employed. An excellent PCA model was obtained with the flow properties of the powder mixture. A good PCA model of tableting performance--based on tablet weight variation and tablet machine speed-was obtained. PMID- 11858522 TI - Development of a topical suspension containing three active ingredients. AB - The objective of this study was to develop a topical suspension that contains sarafloxacin hydrochloride (1 mg/mL), triamcinolone acetonide (1 mg/mL), and clotrimazole (10 mg/mL), and is stable at room temperature (15-28 degrees C) for clinical usage. Due to the difference in the physicochemical properties and chemical stability profiles of these three active ingredients, it is a challenge to develop a stable suspension formulation containing these three drugs. In this study, the stability of these drugs in different buffer solutions was determined under different accelerated isothermal conditions. The Arrhenius equation was subsequently utilized to predict the room-temperature stability of these three drugs in these buffer solutions. By knowing the room-temperature solubility of the drugs in the buffer solution, the stability of the drugs in suspension was predicted. As a result, a 0.02 M phosphate buffer (pH 7.0) containing 0.02% (w/v)polysorbate 20, 1% (w/v) NaCl, and 0.1% (w/v) EDTA was determined to be an acceptable medium. In addition, 0.35% (w/v) high-viscosity carboxymethylcellulose (HV-CMC) was first selected as the suspending agent to enhance the redispersibility of the suspension. Stability data further supported that all three drugs were stable in the suspension containing HV-CMC with less than 5% potency loss for at least 6 months at 40 degrees C and 12 months at 25 degrees C. However, the viscosity drop of this HV-CMC formulation at 25 degrees C and 40 degrees C became a product stability concern. To improve the viscosity stability of the suspension, the medium-viscosity carboxymethylcellulose (MV-CMC) was selected to replace the HV-CMC as the suspending agent. The optimal combination of MV-CMC and sodium chloride in achieving the most desirable dispersion properties for the formulation was determined through the use of a 32 factorial design. The optimal formulation containing 1% MV-CMC and 1% sodium chloride has shown improved viscosity stability during storage and has been used for clinical studies. PMID- 11858523 TI - Polydisperse powder mixtures: effect of particle size and shape on mixture stability. AB - The effect of the shape and size of the components on the stability of mixtures was evaluated in binary mixtures of drug and carrier. Aspirin was used as model drug; spray-dried lactose and microcrystalline cellulose were used as carriers. The coefficient of variation (CV) of the drug in the mixture at various time intervals during mixing was used as a measure of homogeneity. The stability of mixtures was assessed under conditions that were conducive to segregation-in this case, prolonged mixing. The pattern of change in CV with time was analyzed in terms of convective, shear, and diffusive mixing stages. The variation resulting from a change in the shape of the carriers was smaller than that resulting from size differences. The segregation rate constant, calculated on the assumption of a first-order mixing process, was found to be larger in mixtures having components of different shape than in mixtures having components of similar shape. In mixtures of micronized drug and carrier, the pattern of change in the CV of drug with mixing time was attributed to the distribution of agglomerates of micronized drug during convective mixing, followed by shearing of agglomerates and, finally, the distribution of the primary particles during diffusive mixing. Mixtures of non-cohesive powders of similar size and shape behaved like random mixtures of non-interacting components. PMID- 11858524 TI - Preparation and characterization of coenzyme Q10-Eudragit solid dispersion. AB - A solid dispersion of Coenzyme Q10 and Eudragit L 100-55 was prepared using solvent evaporation method. Solid dispersion, physical mixture, and pure compound were then characterized using differential scanning calorimetry and powder x-ray diffraction. Solubility of CoQ10 in different surfactant media was measured, and a suitable dissolution medium was developed to compare the dissolution patterns of the solid dispersion, physical mixture, and the pure compound. Combining labrasol with different surfactants in dissolution media demonstrated an additive effect on CoQ10 solubility. The solubility of CoQ10 in a 4% Labrasol/2% Cremophor EL solution was 562 microg/ml, which was five times higher than the combined solubility in 5% Labrasol (91 microg/ml) and 5% Cremophor EL (7.8 microg/ml). Moderate change in the crystalline pattern of CoQ10 was observed, which was attributed to solvent displacement rather than the degree of crystallinity change. The dissolution test indicated that the in-vitro release of Coenzyme Q10 from its solid dispersion was much faster than its physical mixture, which in turn was faster than the pure drug. The amount of drug released in 12 hours from solid dispersion, physical mixture, and the pure drug was 100, 26.5 and 12.5% respectively. CoQ10 was photostable throughout the dissolution experiments. PMID- 11858525 TI - Elevation of plasma levels of L-dopa in transdermal administration of L-dopa butylester in rats. AB - To increase delivery of L-dopa in its transdermal absorption, a new lipophilic derivative of L-dopa, L-dopa-butylester, was synthesized. An in-vitro study employing two-chamber diffusion cells, in which the excised rat abdominal skin was mounted, revealed that, in the presence of L-menthol and ethanol, L-dopa butylester penetrated in its original form more effectively than L-dopa. L-Dopa butylester sheets were made by immersing wiper sheets in methanol containing the compound, and then evaporating the methanol. An extraction study of the compound from the sheets revealed that its stability was maintained for at least 12 weeks. In an in-vivo cutaneous absorption study, an L-dopa-butylester sheet was attached to the shaved rat abdominal skin. A hydrogel containing L-menthol and ethanol was spread on vinyl tape, and this sheet was placed over it. In plasma, the L-dopa level rose linearly between 30 and 180 min after the cutaneous application; L dopa-butylester was not detected. The L-dopa level was higher than that in which L-dopa was applied. These findings indicated that the lipophilic nature of L-dopa butylester further increased its penetration through the skin, and that L-dopa butylester that was taken up into the general circulation system was rapidly converted to L-dopa by hydrolysis in the body. PMID- 11858526 TI - Application of fluidized hot-melt granulation (FHMG) for the preparation of granules for tableting; properties of granules and tablets prepared by FHMG. AB - The objective of this study was to investigate the properties of granules and tablets prepared by a novel Fluidized Hot-Melt Granulation (FHMG) technique. Macrogol 6000 (polyethylene glycol 6000, PEG 6000), macrogol 20000 (polyethylene glycol 20000, PEG 20000), and glyceryl monostearate (GMS) were used as binders with different levels of viscosity and water solubility. The properties of both granules and tablets were compared with those obtained using the Standard Tablet Formulation (STF, lactose/corn starch/hydroxypropylcellulose/ magnesium stearate: 66/30/3.5/0.5) for fluidized-bed granulation, which is widely used for wet granulation. To obtain suitable flowability as granules for tabletting, the content of the melting material should be approximately 10 w/w%. The rate of increase in the mean diameter of the granules during FHMG was affected by both the melting temperature and the viscosity of the melting material used in the granules. The compression properties of granules prepared by FHMG were also investigated, demonstrating that these granules had a high pressure transmittance. The hardness and the disintegration time of tablets obtained from granules prepared by FHMG were influenced by the properties of the melting material, such as its compaction behavior, solubility, and wettability. No significant differences of hardness were observed when compared to STF tablets. Tablets prepared from FHMG granules disintegrated within 15 min, whereas the STF tablets showed faster disintegration. It was also demonstrated that the hardness and disintegration time of tablets prepared from FHMG granules were not affected by the tablet porosity. Therefore, tablets with a constant quality may be obtainable under a wide range of compression forces. The results of this study suggested that FHMG is a useful method of preparing granules for tableting without using any solvents or water. PMID- 11858527 TI - Novel use of similarity factors f2 and Sd for the development of diltiazem HCl modified-release tablets using a 3(2) factorial design. AB - The objective of this study was to develop modified-release tablets of diltiazem HCl using a direct compression technique. A 3(2) factorial design was employed using the amount of alkali-treated guar gum and cetyl alcohol as independent variables. This article proposes the use of a novel approach-f2 and Sd values as dependent variables-to evaluate the effect of selected independent variables along with other dependent variables (i.e., percentage drug released in x min, Yx; time required for z% drug release, tz; and mean dissolution time (MDT)). It is concluded that when a decision is to be made for the selection of a best batch, it is perhaps more realistic to use the f2 or Sd value which takes into account the dissolution profile as a whole, as opposed to Yx and tz values which use just one point from the dissolution plot. The batch showing the f2 value nearest to 100 or the Sd value nearest to zero is ranked as the best batch (diltiazem HCl 90 mg, alkali-treated guar gum 80 mg and cetyl alcohol 15 mg). The gel strength and matrix erosion of the formulated tablets were dependent on the type and amount of the adjuvants. The drug release rate is well correlated with matrix erosion. The kinetics of drug release fitted best to the Korsmeyer and Peppas model. It is concluded that by using a proper combination of the hydrophilic polymer and cetyl alcohol one can achieve a desirable drug release pattern. PMID- 11858528 TI - Physico-mechanical characterization of policosanol, a novel hypocholesterolemic drug. AB - As part of the formulation studies of policosanol, a new hypocholesterolemic drug, a physico-mechanical characterization was developed. Thermal analysis, using differential scanning calorimetry was used to evaluate the purity of policosanol from batch to batch and, also, the particle size distribution. The degree of wettability of policosanol was studied by measuring the contact angle and solubility in different solvents. The compressibility and cohesion of particles were evaluated using a profile of compression forces, ranging between 6.5 kN and 39.0 kN. Also, other properties such as flow properties, true density, and tapped and bulk density were measured. The industrial batches of policosanol that were studied show an adequate purity and a uniform distribution of the particle sizes. Policosanol shows good flow properties, compressibility, and cohesion as well as a low solubility in the majority of the solvents used in the pharmaceutical industry, and its solubility in water or in aqueous solutions was, mainly, null. The wettability of policosanol in the different solvents shows the following order: methylene chloride > ethanol > acetone >> water. PMID- 11858529 TI - Intramedullary spinal tuberculoma and syringomyelia. PMID- 11858530 TI - Education in the clinical neurosciences. PMID- 11858531 TI - Progress in clinical neurosciences: The neuropathogenesis of HIV infection: host virus interaction and the impact of therapy. AB - Despite the availability of highly active antiretroviral therapy (HAART), primary HIV-related neurological diseases remain major problems in HIV clinics. The present review examines the pathogenesis of HIV-related dementia and the less severe minor cognitive and motor deficit, together with distal sensory and drug induced toxic polyneuropathies. Abnormal host immune responses within the nervous system and the role of viral expression and diversity are emphasized in relation to neurovirulence. Induction of innate immune responses within the central and peripheral nervous systems, largely mediated by cells of macrophage lineage, appear to be common to the development of primary HIV-related neurological disease. Activation of these cell types results in the release of a cascade of inflammatory molecules including cytokines, chemokines, matrix metalloproteinases, and arachidonic acid metabolites that influence neuronal survival. Individual viral proteins encoded by envelope and tat genes and discrete sequences within these genes influence the extent to which these pro inflammatory molecules are induced. At the same time, systemic immune suppression may influence the occurrence and severity of HIV-related neurological diseases. Implementation of HAART and neuroprotective treatments improves neurological function although the evolution of drug-resistant viral strains limits the sustained benefits of HAART. PMID- 11858533 TI - Non-atherosclerotic fusiform cerebral aneurysms. AB - BACKGROUND: Fusiform cerebral aneurysms are dilatations of the entire circumference of a segment of cerebral artery, usually considered due to atherosclerosis in adults. They are relatively thick-walled and elongated, causing neural compression or ischemia when discovered. We have noted a subset of fusiform cerebral aneurysms that vary from this common description. PATIENTS: Out of a series of 472 intracranial aneurysms treated over 11 years, 11 patients between the ages 16 and 67 years (mean age 37) were identified who had discrete fusiform aneurysms unassociated with generalized cerebral atherosclerosis, connective tissue disorder or inflammation. Three presented with hemorrhage, six with neural compression by the aneurysm and two were discovered incidentally. RESULTS: Nine aneurysms were located in the posterior circulation, the other two in the intracranial carotid artery. Their mean length and width were 16.3 and 11 mm, respectively. Three aneurysms contained thrombus. The eight aneurysms that were exposed surgically were partly or substantially thin-walled with normal appearing parent arteries. Eight were treated with proximal occlusion and three were circumferentially "wrapped". Parent artery occlusion caused one death and one mild disability and the remaining patients made good recoveries (follow-up 0.5 - 10 years). CONCLUSIONS: There is a subset of cerebral aneurysms with discrete fusiform morphology, apparently unrelated to cerebral atherosclerosis or systemic connective tissue disease, thin-walled in part or whole, more common in the vertebrobasilar system, and possessing a risk of rupture. Treatments currently available include proximal occlusion or aneurysm wrapping", different approaches than neck-clipping or endovascular coiling of side-wall saccular cerebral aneurysms that leave the parent artery intact. PMID- 11858532 TI - Cluster headache: evidence for a disorder of circadian rhythm and hypothalamic function. AB - This article reviews the literature for evidence of a disorder of circadian rhythm and hypothalamic function in cluster headache. Cluster headache exhibits diurnal and seasonal rhythmicity. While cluster headache has traditionally been thought of as a vascular headache disorder, its periodicity suggests involvement of the suprachiasmatic nucleus of the hypothalamus, the biological clock. Normal circadian function and seasonal changes occurring in the suprachiasmatic nucleus and pineal gland are correlated to the clinical features and abnormalities of circadian rhythm seen in cluster headache. Abnormalities in the secretion of melatonin and cortisol in patients with cluster headache, neuroimaging of cluster headache attacks, and the use of melatonin as preventative therapy in cluster headache are discussed in this review. While the majority of studies exploring the relationship between circadian rhythms and cluster headache are not new, we have entered a new diagnostic and therapeutic era in primary headache disorders. The time has come to use the evidence for a disorder of circadian rhythm in cluster headache to further development of chronobiotics in the treatment of this disorder. PMID- 11858534 TI - Lactate stress testing in 155 patients with mitochondriopathy. AB - OBJECTIVE: Few data are available about the diagnostic yield of the lactate stress test (LST) in a large group of patients with mitochondriopathy (MCP). METHODS: Serum lactate was determined once before, three times during, and once after a 15-minute, constant 30W workload on a bicycle in 62 controls, aged 17 to 84 years, 155 patients with MCP, aged 17 to 87 years, and 31 patients with neurological disorders other than MCP. RESULTS: Lactate's upper reference limits at rest, 5, 10, 15 minutes after starting, and 15 minutes after finishing the exercise were 2.0, 2.1, 2.1, 2.1 and 1.8 mmol/l respectively. The test was regarded abnormal if more than two of the five lactate values exceeded the cut off levels. Among the 103 patients with abnormal LST, 64 (62 %) had normal resting lactate. The sensitivity of the test was 67% and the specificity 94%. CONCLUSION: The LST proved to have a high sensitivity and specificity in the detection of patients with MCP, being thus a simple but powerful tool to assess the impaired oxidative metabolism in MCP patients. PMID- 11858535 TI - Non-atherosclerotic fusiform aneurysms. PMID- 11858536 TI - Carotid dissection: technical factors affecting endovascular therapy. AB - BACKGROUND AND PURPOSE: To delineate factors associated with the successful endovascular treatment of extracranial carotid dissections, the authors review their management of 13 cases. METHODS: The records of 12 patients with 13 dissections were assessed with reference to mechanism of dissection, preoperative symptoms, presence of a pseudoaneurysm, treatment success, and etiology of treatment failure. Patients were followed prospectively and included six men and six women, ranging in age from 27 to 62 years. RESULTS: Angioplasty and stenting were performed successfully in 11 of 13 procedures (10 of 12 patients). Follow-up in these 10 patients demonstrated excellent patency through the stented segment in nine of the 11 treated vessels. Two patients, both of whom suffered their original dissection as a result of endarterectomy, required further angioplasty and stenting for stenosis outside the previously treated arterial segment. Regarding the treatment failures, a stent deployment device could not navigate a tortuous loop in one, while a microwire could not be advanced beyond a pseudoaneurysm in the second. Six patients had pseudoaneurysms, four of which were treated only with stenting across the dissected arterial segment. All pseudoaneurysms treated in this fashion resolved. No permanent complications occurred as a result of endovascular therapy. CONCLUSIONS: Angioplasty and stenting can be performed safely to manage carotid dissection. A pseudoaneurysm or tortuous anatomy can preclude therapy although the former typically resolves if angioplasty and stenting are feasible. Dissections secondary to endarterectomy may be associated with a higher rate of restenosis after stenting and may require further treatment. PMID- 11858537 TI - Invited review: clinical and basic neurophysiology of generalised epilepsies. AB - Electroencephalography (EEG) clarifies several aspects of generalised epileptic seizures and epilepsies. For the clinician, it assists in the diagnosis of the epileptic condition and helps assign the disorder to an appropriate syndrome. This assignation and the quantity of epileptic discharges estimate severity and prognosis. When combined with relevant basic science investigations, EEG studies may disclose significant pathophysiological mechanisms. Therefore, this paper first describes EEG characteristics of the several disorders included under the broad category of "generalised". The review then relates these phenomena to germane experimental data intending that this binocular survey will provide a more meaningful perspective of these disorders. PMID- 11858538 TI - The clinical profile of nonmotor fluctuations in Parkinson's disease patients. AB - OBJECTIVE: Recently described nonmotor fluctuations may cause disability in Parkinson's disease patients. These fluctuations are generally grouped as sensory, autonomic and psychiatric. The clinical spectrum and frequency of these fluctuating symptoms are not well-described. METHODS: We studied the relationship of nonmotor fluctuations with motor symptoms and determined the influence of age at disease onset, duration of disease, dosage and duration of levodopa treatment in the appearance of nonmotor fluctuations. RESULTS: Statistical analysis showed a relationship of disease-related parameters with sensory and autonomic fluctuations but psychiatric fluctuations were only found to be associated with the duration of levodopa usage. The nonmotor fluctuations included in the study were observed during "on" periods as well as "off' periods. CONCLUSION: Nonmotor fluctuations had variable presentations. Moreover, their co-appearance with different types of motor fluctuations may be linked to the effect of other neurotransmitter systems acting synchronously with dopamine. Risk factors for sensory and autonomic fluctuations in patients with Parkinson's disease were early age of disease onset, longer duration and higher dose of levodopa use. Psychiatric fluctuations were only associated with higher doses of levodopa. PMID- 11858540 TI - Tapping and peg insertion after levodopa intake in treated and de novo parkinsonian patients. AB - BACKGROUND: Investigators use instrumental tasks for objective assessment of parkinsonian motor disability and its drug response. To date, such studies on treated parkinsonian patients have not addressed acute and long-term effects of dopaminergic drugs. OBJECTIVES: To determine the impact of long-term dopaminergic therapy within a standardized levodopa challenge test design in combination with two repeatedly performed instrumental tasks, peg insertion and tapping, in previously treated and untreated parkinsonian patients. RESULTS: Tapping significantly deteriorated in previously untreated, but not in treated parkinsonian patients after levodopa intake. In contrast, motor symptoms and peg insertion significantly improved in both groups of parkinsonian patients. Results of both tests differed between parkinsonian patients and matched controls. CONCLUSION: Worsening of cognitively less demanding tapping may result from upregulated presynaptic inhibitory feedback regulation, sedative effects of levodopa or dopamine overflow in untreated parkinsonian patients, who are sensitive to these effects in contrast to treated parkinsonian patients. Tapping is a task with autonomic repetitive performance and programming of standardised movements with a low need for cognitive effort. This autonomic functioning of attentional control and selective processing is intact in Parkinson's disease. Peg insertion depends on more complex movements and thus hypothetically on dopamine-associated cognitive processes. Therefore, impairment of peg insertion responded to dopaminergic stimulation in both groups of parkinsonian patients. Future studies on the efficacy of antiparkinsonian drugs, using instrumental tasks for objective assessment, should consider long-term impact of antiparkinsonian drug therapy and associated cognitive efforts. PMID- 11858539 TI - Respiratory muscle performance and the Perception of dyspnea in Parkinson's disease. AB - BACKGROUND: Pulmonary and respiratory muscle function impairment are common in patients with Parkinson's disease (PD). However, dyspnea is not a frequent complaint among these patients, although it is well documented that the intensity of dyspnea is related to the activity and the strength of the respiratory muscles. PATIENTS AND METHODS: We studied pulmonary function, respiratory muscle strength and endurance and the perception of dyspnea (POD) in 20 patients with PD (stage II and III Hoehn and Yahr scale) before and after their first daily L-dopa dose. Respiratory muscle strength was assessed by measuring the maximal inspiratory and expiratory mouth pressures (PImax and PEmax), at residual volume (RV) and total lung capacity (TLC) respectively. The POD was measured while the subject breathed against progressive load and dyspnea was rated using a visual analog scale. RESULTS: Respiratory muscle strength and endurance were decreased and the POD was increased during the off medication period compared to normal subjects. There was a nonsignificant trend to an increase in Plmax, PEmax and endurance after L-dopa intake. The POD of PD patients decreased (p<0.05) following medication, although, it remained increased (p<0.01) as compared to the normal subjects. Even if patients had spirometry data showing a mild restrictive pattern, before medication, both forced vital capacity (FVC) and forced expiratory volume (FEV)1 remained almost identical after L-dopa intake. CONCLUSIONS: Patients with PD have higher POD, compared to normal subjects and this increased perception is attenuated when the patients are on dopaminergic medication. The change in the POD is not related to changes in respiratory muscle performance or pulmonary functions. A central effect or a correction of uncoordinated respiratory movements by L-dopa may contribute to the decrease in POD following L-dopa treatment. PMID- 11858541 TI - The communication of neurological bad news to parents. AB - Communicating disappointing or unexpected neurological news to parents is often both difficult and emotionally unwelcome. At the same time, it is important that transfer of such information is done well and, indeed, if done well, can be a very rewarding experience. Limited references are available for physicians regarding the proper communication of neurological bad news to parents. This paper attempts to provide general guidelines regarding this process. The review is based on the available medical literature, detailed discussions with many senior physicians from different medical systems and the authors personal experience. The manner in which neurological bad news is conveyed to parents can significantly influence their emotions, their beliefs and their attitudes towards the child, the medical staff, and the future. This review of the literature, combined with clinical experience, attests to the fact that most families describe emotional shock, upset, and subsequent depression after the breaking of news of a bad neurological disorder. However, the majority find the attitude of the news giver, combined with the clarity of the message and the news giver's knowledge to answer questions as the most important aspects of giving bad news. PMID- 11858542 TI - Nipah encephalitis outbreak in Malaysia, clinical features in patients from Seremban. AB - BACKGROUND: An outbreak of viral encephalitis occurred among pig industry workers in Malaysia in September 1998 to April 1999. The encephalitis was attributed to a new paramyxovirus, Nipah virus. This is a description of the clinical features of 103 patients treated in the Seremban Hospital with characterization of the prognostic factors. METHODS: Clinical case records and laboratory investigations were reviewed. The case definition was: patients from the outbreak area, direct contact or in close proximity with pigs, clinical or CSF features of encephalitis. RESULTS: The mean age was 38 years, 89% were male, 58% were ethnic Chinese, 78% were pig farm owners or hired workers. The mean incubation period was 10 days. The patients typically presented with nonspecific systemic symptoms of fever, headache, myalgia and sore throat. Seizures and focal neurological signs were seen in 16% and 5% respectively. In the more severe cases, this was followed by drowsiness and deteriorating consciousness requiring ventilation in 61%. Autonomic disturbances and myoclonic jerks were common features. The mortality was high at 41%. Systolic hypertension, tachycardia and high fever were associated with poor outcome. On the other hand, 40% recovered fully. As for the other 19%, the residual neurological signs were mostly mild. CONCLUSION: Nipah virus caused an encephalitis illness with short incubation period and high mortality. The prognosis for the survivors was good. PMID- 11858543 TI - Global cerebral anoxia. PMID- 11858545 TI - Idiopathic free-floating thrombus of the common carotid artery. AB - BACKGROUND: The observation of an intraluminal common carotid artery thrombus overlying a wall defect at ultrasonography or angiography is unusual. To our knowledge, there are no previous reports of a free-floating thrombus in the common carotid artery. CASE REPORT: A 45-year-old woman who was previously healthy and on no medications presented with acute hemiparesis and aphasia. Following testing that included carotid duplex and transcranial Doppler ultrasonography, diffusion-weighted magnetic resonance imaging, and digital subtraction angiography, the patient underwent emergency open embolectomy. No underlying wall defect was seen at the time of imaging or surgery. No obvious hypercoagulable state could be identified. Her NIH Stroke Scale score improved from 26 at admission to 2 at three months and 1 at one year. CONCLUSIONS: Multimodal imaging may have improved diagnosis and management in this patient with a unique finding. The source of the thrombus remains obscure. PMID- 11858544 TI - New insights into the neuropathogenesis of molybdenum cofactor deficiency. AB - BACKGROUND: Molybdenum cofactor deficiency (MOCOD) is a rare, progressive neurodegenerative disorder caused by sulphite oxidase enzyme deficiency. The neuropathological findings are consistent with a toxic insult to the brain that causes severe neuronal loss, reactive astrogliosis and spongiosis. The mechanisms responsible for these changes are unknown. METHODS: The case is a male infant with MOCOD who died at nine months of age from pneumonia. At autopsy, a complete neuropathological examination was performed including conventional immunohistochemical staining. In addition, brain sections were stained cytochemically with shikata and orcein which stain for disulphide bonds. The elemental composition of cortical cells was then analyzed in the scanning electron microscope using backscatter electron imaging and energy dispersive X ray spectrometry. RESULTS: Neurons demonstrated cytoplasmic staining with shikata and orcein cytochemically when compared to control sections. Energy dispersive X ray spectrometry analysis of these neurons confirmed the presence of excess sulphur and unexpectedly revealed excess magnesium accumulation. None of these findings was found in an age-matched control. CONCLUSIONS: In MOCOD we found abnormal accumulation of sulphur and magnesium in neurons. It is postulated that sulphur-containing compound(s) that are formed as a result of MOCOD cause excitotoxic neuronal injury in the presence of excess magnesium. PMID- 11858546 TI - Iso-N-formyl-5-en-chonemorphine, a steroidal alkaloid from Sarcococca zeylanica. AB - Chemical investigation of the non-quaternary alkaloidal fraction of the aerial parts of Sarcococca zeylanica of the family Buxaceae furnished a steroidal alkaloid iso-N-formyl-5-en-chonemorphine, which has not been previously reported as a natural product. The structure of this alkaloid was established on the basis of spectroscopic evidence. PMID- 11858547 TI - Two new triterpenoid saponins from Aralia subcapitata. AB - Two new oleanane-type saponins, subcapitatoside B and C were isolated from the roots of Aralia subcapitata. On the basis of chemical and spectral evidences, subcapitatoside B and C were established as oleanolic acid 3-O-beta-D glucopyranosyl-(l-->3)-[beta-D-galactopyranosyl- (1-->2)]-beta-D3- galactopyranoside, and 3-O-beta-D-glucopyranosyl- (1-->3)-[beta-D galactopyranosyl-(1-->2)]-beta-D-glucopyranosyl oleanolic acid 28-O-beta-D glucopyranosyl-(1-->6)-beta-D-glucopyranoside, respectively. PMID- 11858549 TI - Headspace constituents of Parkia speciosa seeds. AB - The headspace of Parkia speciosa seeds was analysed by means of GC and GC-MS and found to contain 21 volatile components. The main constituents were hydrogen sulphide (41.30%), ethanol (39.15%), 1,2,4-trithiolane (4.75%) and acetaldehyde (3.59%), of which 1,2,4-trithiolane was found as one of the main component for the characteristic odour. PMID- 11858548 TI - Asaricin, the main component of Ocotea opifera Mart. essential oil. AB - Investigation of the volatile fraction from the stem bark of Ocotea opifera Mart. led to the isolation and characterization of asaricin, a phenolic derivative with antifungal and insecticidal activity, as the main component, which is described for the first time for the genus Ocotea. The structure has been established by a study of its mono- and bidimensional NMR spectra and mass spectrometry. PMID- 11858550 TI - Triterpenes from Mimusops elengi. AB - Pentacyclic triterpenes (1) and (2) have been isolated from Mimusops elengi and assigned structures 3beta,6beta,19alpha,23-tetrahydroxy-urs-12-ene and 1beta hydroxy-3beta-hexanoyllup-20 (29)-ene-23, 28-dioic acid, respectively, on the basis of spectroscopic studies including 2D-NMR. The compound 1 showed moderate inhibiting activity against beta-glucuronidase enzyme PMID- 11858551 TI - MAP: microwave-assisted extraction of fatty acids and Py/GC/MS analysis of selected insects. AB - A Microwave-Assisted Process (MAP) solvent extraction procedure was used in conjunction with GC/MS analysis to investigate the chemical composition of dried silkworm, dried earthworm, silkworm droppings, and cicada nymph skin. Selected insect samples were also analyzed by pyrolysis-GC-MS. Silkworm and earthworm generated fatty acids, sterols and phenol derivatives. Cicada nymph skin was analyzed by pyrolysis/GC/MS and generated mainly heterocyclic compounds. The data indicated that insects can retain environmental contaminants such as 2,6-bis(1,1 dimethylethyl)-4-methyl phenol (BHT) and as such could be used as biological indicators. In addition, some of the therapeutic characteristics associated with insects could be attributed to the chemicals ingested from plant sources. PMID- 11858552 TI - Microwave-assisted extraction of phenolic compounds from grape seed. AB - A microwave-assisted extraction technique was developed to optimize the extraction of phenolic compounds from grape seeds. The microwave power (300-150W) and time of extraction (20-200s) were varied during the optimization process. The polyphenol content of the resulting extracts were measured as mg of tannic acid equivalent per gram of crude extract (mg TAE/g of crude extract), using a Folin Ciocalteau reagent. In general, neither the time nor the power had a significant effect on the overall % yield (average of 13.5%) and on the polyphenol content (392 mg TAE/g of crude extract) of the extracts. However, when the solvent polarity was changed by the addition of 10% water, the yield increased to 15.2% and the polyphenol content increased to 429 mg TAE/g of crude extract. PMID- 11858553 TI - Inhibition of acetylcholinesterase activity by essential oil from Citrus paradisi. AB - Inhibition of acetylcholinesterase (AChE) activity by essential oils of Citrus paradisi (grapefruit pink in USA) was studied. Inhibition of AChE was measured by the colorimetric method. Nootkatone and auraptene were isolated from C. paradisi oil and showed 17-24% inhibition of AChE activity at the concentration of 1.62 microg/mL. PMID- 11858554 TI - Creating healing environments in critical care units. AB - The concept of healing environments is not new. The idea of creating an environment that would facilitate healing was popular over 2000 years ago, but it has been only recently that this concept has been applied to the design of hospitals and specifically to intensive care units. Although it is necessary to address the needs of the professional staff who are caring for seriously ill patients, it is also critical to include specific elements that will enhance the patients' healing process. It has been demonstrated that views of nature, natural light, soothing colors, therapeutic sounds, and the interaction of one's family can enhance the healing process. These elements important to a patient's healing process must be considered and balanced with the needs of health care providers in the design of critical care environments. PMID- 11858555 TI - Designing humanistic critical care environments. AB - The critical care environment can be designed to become more humanistic. Consideration of the environmental challenges of noise, lights, color, views, temperature, and comfort is essential. This article identifies the issues and concerns in the design of more humanistic healing in critical care units. Strategies to improve the environment include improving the physical and emotional tone of the unit through creative design, family and pet visitation, sleep promotion, and aromatherapy among others. In a life-threatening illness, attention paid to these concerns may significantly improve quality of life for patients and family. PMID- 11858556 TI - Critical care unit design: a nursing perspective. AB - The task of designing a new critical care unit is best accomplished with the input of people representing multiple disciplines including architects, engineers, physicians, nurses, and equipment manufacturers. It is imperative that the critical care nursing staff and management take an active role in planning the layout of the unit and patient rooms, as the nurses will be the bedside providers 24 hours a day. The new unit should be designed to offer efficient patient care as well as a healing, comfortable environment for both the patients and their families. PMID- 11858557 TI - Infection control considerations in critical care unit design and construction: a systematic risk assessment. AB - When contemplating major renovation or new construction of a critical care unit (CCU), the use of systematic infection control risk assessment (ICRA) provides guidance to limit infectious perils for patients and to reduce occupational hazards for employees in this environment. The nursing representative and other members of the multidisciplinary planning and design team must routinely address infection control factors throughout the project and assist administration in understanding the rationale for the floor plan, equipment, and furnishings required to support sound infection control practices. Collaborative team skills, articulate communication techniques, and frequent rounds are integral throughout the construction. PMID- 11858558 TI - Streamlining patient care processes through flexible room and equipment design. AB - Hospitals are planning and designing new environments to accommodate a greater concentration of critically ill patients and to permit the installation of sophisticated technology required for the care of this high-acuity population. A serious nursing shortage, customer demands for privacy and personal amenities, and cost constraints pose numerous problems in planning and design processes. The flexible, acuity-adaptable room concept has been introduced as a platform for changing care processes to achieve a cost-effective care model that fosters nursing efficiency and increased customer satisfaction. PMID- 11858559 TI - Managing medication errors by design. AB - It is estimated that over 7,000 U.S. citizens perish each year from medication errors. The critically ill are among the patient group at highest risk for a medication error. Knowledge is the key to error prevention. Applying information such as risk factors for error commission, new regulatory standards, and approaches to error management will maximize your patients' and your own safety. PMID- 11858560 TI - The disease management approach to controlling asthma. AB - Asthma has become an important public health issue worldwide and certain groups, such as children, are at particular risk of the disease. Often asthma remains under-diagnosed and under-treated. Despite these worrying trends, the disease management approach to asthma control can help most asthma patients achieve a 'normal' way of life. The increased prevalence and greater diagnostic awareness of asthma have placed increased demands on healthcare resources, but effective asthma control can minimize the personal, social and economic burdens of asthma. Early diagnosis and immediate anti-inflammatory treatment is the first step in gaining control of symptoms. A stepwise approach is then used to classify asthma severity and treatment, with the number and frequency of medications increasing (step up) as asthma severity increases and decreasing (step down) when asthma is under control. This stepwise approach to asthma management necessitates regular review of treatment once asthma is under control. However, effective asthma management is dependent on successful patient education, adherence to prescribed medication and good doctor patient partnerships. Current treatment guidelines recommend the use of a written asthma management plan that should be agreed between the doctor and patient. These plans should cover all aspects of asthma treatment, including prevention steps for long-term control and action steps to stop attacks once a worsening in asthma has been recognized. This comprehensive approach to asthma management increases the likelihood of achieving asthma control, which in turn reduces the need for emergency visits to the hospital or clinic and reduces the limitations on physical activity previously imposed by the condition. PMID- 11858561 TI - Symbicort: controlling asthma in adults. AB - Inhaled corticosteroids reduce airway inflammation and bronchial hyper responsiveness in asthma. Their regular use is associated with a reduction in morbidity and mortality from asthma. International guidelines recommend their use as first-line treatment in asthma and the dose should be increased stepwise in accordance with asthma severity. Short-acting beta2-agonists are recommended for use as reliever medication for the acute relief of symptoms. Long-acting beta2 agonists have a sustained bronchodilator action but are not recommended for use as monotherapy in asthma, particularly because they lack clinically relevant anti inflammatory action. The seminal observation in a controlled trial that the addition of salmeterol to low-dose inhaled beclomethasone dipropionate was superior to doubling the dose ofthe beclomethasone dipropionate was followed by several well-controlled studies that confirmed this observation. These included multinational and multicentre trials in large numbers of patients and the use of different inhaled corticosteroids and the two long-acting beta2-agonists, formoterol and salmeterol. Therefore level A evidence (randomized controlled trials with a rich body of evidence) is available for this recommendation. Based on this, updated asthma guidelines recommend the addition of long-acting beta2 agonists for chronic asthma that is at least moderately severe or mild asthmathat is not well controlled with low-dose inhaled corticosteroids. Attention was therefore focused on developing combination inhalers that included inhaled corticosteroids and long-acting beta2-agonists in order to simplify asthma treatment and improve adherence to prescribed medication. Symbicort Turbuhaler (budesonide/formoterol in a single inhaler) is one such development. Studies to date in over 800 adult patients have confirmed that budesonide/formoterol is superior to double the dose of budesonide monotherapy and at least as effective and safe as budesonide and formoterol in separate inhalers, These studies also demonstrate that there are no physico-chemical interactions when the two medications are combined in a single inhaler. Symbicort in its easy to use formulation and inhaler device represents a valuable addition to the pharmacological management of asthma. PMID- 11858562 TI - Symbicort: controlling asthma in children. AB - Asthma is now widely recognized as a chronic inflammatory condition of the airways that requires early pharmacological treatment and long-term management. Anti-inflammatory agents, particularly inhaled corticosteroids, are currently the most effective long-term preventative medication. Moreover, early intervention with inhaled corticosteroids plays an important role in airway remodelling. Despite significant advances in the understanding of asthma and its pharmacological management, the prevalence of asthma in children, teenagers and young adults is on the increase. For patients whose asthma is not fully controlled with daily inhaled corticosteroid therapy, national guidelines advocate the addition of long-acting inhaled beta2-agonist therapy, rather than an increase in dose of inhaled corticosteroids, for the treatment of persistent childhood asthma. However, adherence to treatment with asthma medication declines as the regimen becomes more complicated. Adherence to therapy and therapy convenience are key to the successful pharmacological management of asthma, particularly in children. The administration of prescribed medication via a single inhaler offers a convenient treatment regimen that has the potential to improve adherence to treatment. This paper presents data to show that the combined administration of budesonide and formoterol via a single inhaler (Symbicort Turbuhaler) is effective and well tolerated in the treatment of asthma in children. PMID- 11858563 TI - The whole story: treatment outcomes with Symbicort. AB - Asthma is a chronic inflammatory disorder of the airways that has a considerable socioeconomic impact. Asthma management guidelines have been introduced to help provide better long-term control of asthma. Although recommended guidelines may increase the direct medication costs, the overall direct costs of asthma may be reduced due to fewer exacerbations. In addition, indirect costs due to lost productivity and mortality are reduced and patients have an improved quality of life. Inhaled corticosteroids are first-line therapy in the treatment of persistent asthma. Against this background, we have assessed the cost effectiveness of Symbicort (budesonide and formoterol in a single inhaler), a treatment that provides better control of asthma compared with budesonide alone. While the prescribing costs of Symbicort were found to be higher than for budesonide alone, these were partially offset by reduced costs due to fewer asthma exacerbations and a reduced need for other medications. Combined long-term therapy with budesonide and formoterol also improves patient quality of life compared with budesonide alone. Two other factors associated with asthma treatment success and cost-effectiveness are patient/physic an education and good patient adherence to prescribed therapy. The introduction of a single inhaler that is easy to use in simple treatment regimens may improve patient adherence to prescribed medication, thus resulting in improved asthma control and fewer exacerbations. Treatment with Symbicort is more cost-effective than treatment with budesonide alone. PMID- 11858564 TI - The role of inflammation and anti-inflammatory medication in asthma. AB - Asthma is a chronic inflammatory disease of the airways involving a wide range of cells and mediators. First-line therapy of persistent asthma involves the use of inhaled corticosteroids to control the underlying inflammation of the airways. Inhaled beta2-agonists are also widely used in asthma therapy and are the most effective bronchodilators currently available. The short-acting beta2-agonists are now used on an as-needed basis for rapid relief of symptoms, In recent years, long-acting inhaled beta2-agonists have had an increasing role in the management of asthma, particularly in patients with moderate to severe asthma. This class of drug has a long duration of action and is recommended as add-on treatment to inhaled corticosteroids in the long-term control of asthma. New therapies have been added to asthma therapy as our understand ng of the pathogenesis of asthma has increased. The use of multiple therapies necessitates a clear understanding of the mode of action of the drugs and any potential interaction or overlap of effect. For many people asthma is associated with complex therapy; thus treatment developments that simplify asthma treatment are an mportant step forward n asthma management. PMID- 11858565 TI - Lateralization of the inferior alveolar nerve with simultaneous implant placement: a modified technique. AB - PURPOSE: Several nerve repositioning techniques have ben presented in the literature, each with limitations. This article presents a new technique involving the use of 2 osteotomies, with minimizes particularly the potential duration of sensory disruption and the risk of nerve paresthesia and inadvertent nerve transection or compression. MATERIALS AND METHODS: Ten patients ranging in age from 47 to 67 years were selected for nerve lateralization utilizing the modified technique. A total of 23 cylindrical implants were placed. An average follow-up period was 29.8 months. RESULTS: Of the 10 patients, 4 experienced total return of sensation within 3 to 4 weeks. One patient experienced complete recovery at 6 weeks. DISCUSSION: Creating 2 osteotomies as described minimizes the chances for postoperative neuropraxia and nerve paresthesia or anesthesia. CONCLUSION: When there is moderate-to-severe bone resorption of the mandible posterior to the mental foramen, repositioning the inferior alveolar nerve using both an anterior and posterior osteotomy allows for more bone to accommodate ideal placement and greater length of implant. PMID- 11858566 TI - Cross-sectional imaging of the jaws for dental implant treatment: accuracy of linear tomography using a panoramic machine in comparison with reformatted computed tomography. AB - PURPOSE: Although various panoramic X-ray machines with linear tomographic functions are now frequently applied to diagnosis related to dental implant treatment, the angles of the tomographic objective planes are automatically determined and cannot be adjusted for individual patients. To resolve this problem, a direct laser positioning (DLP) system was developed. In this investigation, the measurement accuracy of images obtained by the DLP system in comparison with those from reformatted computed tomography (CT) was assessed. MATERIALS AND METHODS: A rectangular parallel piped phantom was scanned with the system and the height and width were measured on linear tomograms. Ten sites in 3 dried mandibles and 21 mandibular molar sites in 15 patients were examined both with the DLP system and the reformatted CT to compare the measured values on both images. RESULTS: The phantom experiment showed that the difference between the actual and measured heights and widths of the phantom were within 1 mm. DISCUSSION: The difference between the values obtained by the DLP system and CT was slightly larger in the patients than those in the dried mandibles. CONCLUSION: The DLP measurement accuracy was deemed sufficient for clinical use. PMID- 11858567 TI - Immediate implant placement following a modified trephine/osteotome approach: success rates of 116 implants to 4 years in function. AB - PURPOSE: A technique is presented which utilizes a trephine with a 3.0-mm external diameter followed by an osteotome to implode a core of maxillary posterior alveolar bone prior to immediate implant placement. MATERIALS AND METHODS: The technique and its indications and contraindications are described in detail. RESULTS: One hundred sixteen implants were placed and uncovered utilizing this technique. Two implants were mobile at the time of uncovering. DISCUSSION: One hundred fourteen implants were restored and have been functioning successfully for up to 4 years according to the Albrektsson criteria, yielding a success rate of 98.3%. CONCLUSION: No implants have been lost or are failing in function. PMID- 11858568 TI - Clinicoanatomic study on the craniofacial bones used for cranio- and maxillofacial implants. AB - PURPOSE: To clarify basic clinico-anatomic factors associated with cranio maxillofacial rehabilitation using implants. MATERIALS AND METHODS: Morphometrically evaluated were items such as the length and width of bone and the thickness of cortical bone at important sites for implant placement into cranial and maxillofacial bones in 30 cadavers at autopsy. RESULTS: At sites corresponding to potential placement sites for implants as the fixation source for ocular epitheses, the mean length of bone was 7.8 mm, and the mean width was 8.3 mm. CONCLUSION: Useful data have been obtained for the selection of the placements sites, direction, length, and external diameter of cranio- and maxillofacial implants. PMID- 11858569 TI - A pause to reflect. PMID- 11858570 TI - In vivo registration of force development with ceramic and acrylic resin occlusal materials on implant-supported prostheses. AB - PURPOSE: It has been hypothesized that the shock generation on implant-supported prostheses during chewing should generate higher implant loads if the veneering material is porcelain rather than acrylic resin. MATERIALS AND METHODS: The present study uses strain-gauged abutments to measure the force transferred to the implant after a shock has been applied. This was measured in vitro and in vivo in 5 patients. RESULTS: The different occlusal materials did not lead to different forces generated to the implants of the patients. DISCUSSION: From a practical point of view, the choice of occlusal material has no bearing per se on force generation to the implants. CONCLUSIONS: The present study demonstrated that there: (a) is a difference in resilience between acrylic resin and ceramic veneering materials, but (b) this difference is only measurable in in vitro where the force is generated by a shock only and the implant is rigidly anchored. PMID- 11858571 TI - Removal torques of conical, tapered implant abutments: the effects of anodization and reduction of surface area. AB - PURPOSE: To examine the effects of anodization (surface coating) and reduction of internal Morse taper surface area on the reversal torque values of Straumann ITI dental implants and abutments. MATERIALS AND METHODS: Eighty ITI solid screw implants were mated with corresponding 5.5-mm solid abutments. The assemblies were divided into 4 test groups of 20 specimens. All abutments were torque tightened into the implant to 35 Ncm. Half of the abutments were anodized and half were in their as-machined state. Each of these 2 groups included half of the implants with the standard internal Morse taper configuration and half with the synOcta (Straumann USA, Waltham, MA) internal positioning interface (indexed). Torque removal testing was then performed on the assemblies. The 4 groups were compared statistically to examine the effect of the 2 variables (anodization and reduction in surface area). Scheffe's test for multiple comparisons was used to compare groups at an adjusted significance level of < or = .05. RESULTS: Torque removal of all specimens revealed that the indexed implant with the non-anodized abutment demonstrated superior removal torque. DISCUSSION: The indexed and standard implants with anodized abutments, and the standard implant with the non anodized abutment had lower reversal torque values. CONCLUSION: The addition of the indexed internal surface to the ITI implant did not have deleterious effect on the resistance to loosening of standard solid abutments. PMID- 11858572 TI - Titanium alloy osseointegration in cancellous and cortical bone of ovariectomized animals: histomorphometric and bone hardness measurements. AB - PURPOSE: Histomorphometry and microhardness measurements were performed after Ti6AI4V implantation in cancellous and cortical bone of healthy and ovariectomized animals to determine characterization of the bone-biomaterial interface in osteopenic bone. MATERIALS AND METHODS: Nine ovariectomized and 9 sham-aged rats were used. Four months later, nails were implanted in the distal femurs, and the animals were sacrificed after 8 weeks. Moreover, 3 ovariectomized and 3 sham-aged sheep were used. Twenty-four months later, screws were implanted in the tibial diaphyses and the animals were sacrificed after 12 weeks. RESULTS: Histomorphometry showed the development of osteopenia in both trabecular and cortical bone, and revealed a significant decrease in the osseointegration rate in osteopenic versus sham-aged animals for both trabecular (Affinity Index: 18.6%, P < .001) and cortical bone (Affinity Index: -23.5%, P < .005; Bone Ingrowth: -9%, P <.05). At the interfaces of the sham-aged animals in both trabecular and cortical bone, a decrease of bone microhardness was observed in comparison with preexisting bone (trabecular: -9.8%, P < .0005; cortical: -19.3%, P < .0005). In case of osteopenia, this decrease was even more extensive (trabecular: -15.5%, P < .0005; cortical: -24.7%, P < .0005). DISCUSSION: The present data suggest that bone formation around Ti6AI4V was not associated with complete bone maturation, even in healthy animals. In case of osteopenia, both bone formation and maturation were delayed. CONCLUSION: These results apparently demonstrate the utility of investigating biomaterials in osteopenic bone and the importance of careful evaluation of the healing rate and bone maturation degree around implanted biomaterials. PMID- 11858573 TI - Association of calcitonin receptor gene polymorphism with early marginal bone loss around endosseous implants. AB - PURPOSE: This study investigated the relationship between calcitonin receptor (CTR) genotype and buccal marginal bone loss observed at stage II surgery for endosseous implants. MATERIALS AND METHODS: A total of 237 implants were placed in 35 patients; 89 implants were placed in maxillae and 148 implants in mandibles. The CTR genetic polymorphism was examined by the polymerase chain reaction restriction fragment length polymorphism method. RESULTS: Patients with the TC genotype were 20 times more likely to suffer buccal marginal bone loss in the mandible than patients with the CC genotype. Furthermore, there were no significant differences in the distribution of age, smoking status, postmenopausal women, and bone quality between patients with and without bone loss in either jaw. DISCUSSION: These results suggest that the known risk factor for bone loss cannot explain the early marginal bone loss around the implants. CONCLUSION: Although further genetic research should be conducted, it is suggested that the CTR genetic test could become a useful tool in the planning of treatment before implant surgery and lead to more predictable implant treatment. PMID- 11858574 TI - Histologic and histomorphometric evaluation of peri-implant bone subjected to immediate loading: an experimental study with Macaca fascicularis. AB - PURPOSE: Immediately loaded splinted implants can become osseointegrated when they are placed in the anterior part of the mandible. The concept of immediate loading has not been well examined in the posterior mandible. The aim of this study was to evaluate the hard tissue reactions around immediately loaded implants placed in the posterior mandible in the monkey model. MATERIALS AND METHODS: Six adult M. fascicularis monkeys were used in this study. Thirty-six Ankylos implants (Degussa Dental, Hanau-Wolfgang, Germany) were placed after extraction of the second premolar, first, and second molar teeth and complete healing of the sockets. Control (C) group implants were placed and, after osseointegration, were loaded for 1 month using temporary acrylic resin prostheses and later for 2 months using splinted metal crowns. In the contralateral region of the mandible, test (T) group implants were placed and loaded immediately with the same sequence as carried out for the C implants. After sacrifice of the animals, specimens were examined histologically and evaluated histomorphometrically. RESULTS: All implants were osseointegrated. Compact, cortical bone in contact with the implant surface without any gaps or connective tissue formation was demonstrated. DISCUSSION: Histomorphometric findings of the bone-implant-contacts showed no significant differences between the T and C group implants. Peri-implant mineralized bone areas presented statistically significant differences and showed a higher density of bone between the threads of immediately loaded implants (P < .05). CONCLUSIONS: Immediately loaded splinted implants in the posterior mandible can become osseointegrated with a hard tissue peri-implant response similar to that of delayed loaded implants. Moreover, immediate loading seems to increase the ossification of the alveolar bone around endosseous implants. PMID- 11858575 TI - Anterior maxillary alveolar distraction osteogenesis: a prospective 5-year clinical study. AB - PURPOSE: Anterior maxillary alveolar vertical distractions were followed for a 5 year period of time. MATERIALS AND METHODS: A total of 30 vertical distractions were done in 28 patients. Two patients had both anterior maxilla and anterior mandibular distractions for a total of 30 distractions. Two distraction techniques were used: an implant device (3i) and an orthodontic screw device (Osteomed) for orthodontic attachment. Both devices enabled some horizontal as well as vertical movement. The average net vertical distraction was 6.5 mm, but the average anterior horizontal movement was less than 2 mm. RESULTS: Eighty-four implants were placed, but 8 implants failed to integrate. DISCUSSION: All failed implants had been placed in poor quality bone that needed bone grafting. The most common restoration was a fixed prosthesis supported by implants; the longest follow-up post loading was 4.4 years. CONCLUSION: This clinical study gives additional evidence in favor of the stability and utility of vertical distraction procedures in the maxillary esthetic alveolar zone. PMID- 11858577 TI - Single-tooth replacement with the Frialit-2 system: a retrospective clinical analysis of 146 implants. AB - PURPOSE: This study was intended to provide a report of experience and results with Frialit-2 implants used for single-tooth replacement. MATERIALS AND METHODS: Over a 7-year period (1994-2000), 146 single-tooth implants (84 maxilla, 62 mandible) were placed in 112 patients (67 females, 45 males; 31.2 +/- 16.4 years). The sites included maxillary anterior teeth (n = 38) as well as the mandibular premolars and molars (n = 57). Ninety-three crowns were cemented and 53 crowns were screw mounted (22 with vertical, 31 with horizontal screws) on standard abutments. The follow-up time varied between 3 and 80 months (35.8 +/- 16.5 months). RESULTS: Two implants (1.4%) were lost, 1 during early loading and the other after 6 years. The most frequent prosthetic complication was isolated crown loosening of cemented crowns requiring recementation of 9 crowns (9.9%). Crowns with vertical screws showed no crown and/or screw loosening. Four crowns (2.8%) were replaced because of ceramic fracture. DISCUSSION: Peri-implant soft tissue condition, bone resorption, and Periotest values indicated satisfactory results. The cumulative implant survival rate during the follow-up period was 97.3%, and that of the crowns 96.4% (total cumulative survival rate 93.7%). CONCLUSIONS: With the low number of abutment screw loosenings (3.5%), the deep internal hexagonal retention compared favorably to external retention methods. The predominant use of long implants (98.4% > or = 13 mm) allowed a favorable implant/crown ratio with the potential for problem-free, long-term results. PMID- 11858576 TI - The influence of mandibular dentition on implant failures in bone-grafted edentulous maxillae. AB - PURPOSE: To evaluate the influence of mandibular dentition on the performance of maxillary implants prior to definitive prosthesis attachment in maxillae that have been reconstructed with autogenous bone grafts. MATERIALS AND METHODS: A retrospective review of 90 consecutive patients, 31 men and 59 women, with a mean age of 57.4 years, was conducted. All patients underwent treatment planning to receive endosseous implants in the edentulous maxilla in conjunction with autogenous bone grafting. During the time from implant and/or bone graft placement until placement of the definitive restorations in the maxillae, the mandibular dentitions were recorded and categorized into 6 groups based upon the presence and distribution of teeth. RESULTS: Of 643 implants placed, 118 (18.4%) were lost between implant placement and definitive prosthesis placement. The type of mandibular dentition was significantly associated with implant failure during this time interval (P < .001). In particular, the patients with implants opposing unilateral occlusal support showed the highest rate of implant failure (43.8%, or 28 of 64 implants). Implants that opposed a mandibular implant-supported fixed prosthesis demonstrated an implant failure rate of 14.3% (10 of 70), and in patients with a removable mandibular denture, the implant failure rate was 6.2% (4 of 65 implants failed). The overall mean patient follow-up was 64.2 months. At 60 months, the cumulative implant failure rate based on the Kaplan-Meier method was 20.2%. DISCUSSION: Unfavorable concentration of forces on the maxilla may contribute to increased risk of implant failure. CONCLUSION: Effort should be made to create a favorable occlusion in the mandible, with attention being paid to broad distribution of occlusal contacts. PMID- 11858578 TI - Use of particulate dentin-plaster of Paris combination with/without platelet-rich plasma in the treatment of bone defects around implants. AB - PURPOSE: To evaluate the effect of particulate dentin-plaster of Paris with and without platelet-rich plasma (PRP) on bone healing and new bone formation around titanium dental implants in a canine model. Histologic sections and histomorphometric analysis of the defects were obtained at 6 and 12 weeks after surgery. MATERIALS AND METHODS: Three circular bone defects were surgically prepared in iliac crest sites in each of 10 animals. A total of 30 Avana dental implants were placed in the animals. They were self-tapping, screw-type implants, 10 mm in length and 4 mm in diameter, all made of commercially pure titanium. A titanium implant was placed centrally in each defect. In each dog, the defects were treated with 1 of the following 3 treatment modalities: (1) no treatment (control); (2) grafting with particulate dentin-plaster of Paris; (3) grafting with particulate dentin-plaster of Paris and PRP. RESULTS: Histologic analysis showed that all of the bone defects surrounding the implants that were treated with particulate dentin-plaster of Paris, with and without PRP, were filled with new bone. The defects that were not treated (control) demonstrated new bone formation only in the inferior threaded portion of the implants. DISCUSSION: Histomorphometric results revealed a higher percentage of bone contact with particulate dentin-plaster of Paris and PRP compared to the control and particulate dentin-plaster of Paris. CONCLUSIONS: These results suggested that bone defects around titanium implants can be treated successfully with particulate dentin-plaster of Paris, and that the outcome can be improved if PRP is also used. PMID- 11858579 TI - Method for the placement of palatal implants. AB - PURPOSE: Palatal implants have been used in the last 2 decades to eliminate headgear wear and to establish stationary anchorage. The aim of this investigation was to establish a method and easy protocol for palatal implant placement. MATERIALS AND METHODS: The study comprised 8 male and 15 female patients each having a 4.5 x 8-mm stepped screw titanium implant placed in the palatal region for orthodontic purposes. A surgical template containing metal drill housing was prepared. Angulation of the drill housing was controlled according to the radiologic tracing of the maxilla transferred to a plaster cast section in the paramedian plane. Implants were placed using a noninvasive technique (incision, flap, and suture elimination) and left transmucosally to facilitate the surgical procedure and reduce operations. The paramedian region was selected so as to avoid connective tissues of the palatine suture and because it was considered to be a suitable host site for implant placement. RESULTS: After 3 months of healing, all implants were osseointegrated and no implant was lost throughout the orthodontic treatment. DISCUSSION: Palatal implants can be used effectively for anchorage maintenance and space-gaining procedures. CONCLUSION: Usage of a 3-dimensional surgical template eliminated faulty implant placement, reduced chair time, and minimized trauma to the tissues while enhancing osseointegration. PMID- 11858580 TI - Keeping nurses in their place? PMID- 11858581 TI - Spiritual assessments in primary care. PMID- 11858582 TI - Encountering cat-scratch disease. PMID- 11858583 TI - Drug and device approval highlights from 2001. AB - The U.S. Food and Drug Administration approved more than 100 drugs and devices in 2001. This article discusses several of the new approvals, including ophthalmic drugs, contraceptive therapies, mental health medications, and medical devices. PMID- 11858584 TI - How to diagnose malignant melanoma. AB - The incidence of malignant melanoma has increased dramatically in recent decades. Although curable if caught early, malignant melanoma often presents a diagnostic challenge. This article discusses modalities for identifying malignant melanoma, including four techniques used in skin examinations: (1) ABCD method, (2) total body photography, (3) skin surface microscopy, and (4) machine vision. PMID- 11858585 TI - Improving your cultural awareness with culture clues. AB - When the University of Washington Medical Center (UWMC) administered a survey to gauge patient education support needs among staff, the results indicated a need for cultural competency. Using action methodology and needs assessment, the committee charged with developing a tool to meet this need launched a new concept: Culture Clues. These brief documents provide culture-specific patient information, allowing staff to approach patients according to their culture. The committee created six Culture Clues and is developing more. PMID- 11858586 TI - Responsibility for your actions. PMID- 11858587 TI - Demystifying trichotillomania. PMID- 11858588 TI - Ring removal. PMID- 11858589 TI - Antibiotic approval. PMID- 11858590 TI - Lennox-Gastaut treatment. PMID- 11858591 TI - New, non-NSAID alternative. PMID- 11858592 TI - A job description for the oncology NP. PMID- 11858593 TI - Unruffling the mystique of therapeutic touch. TT: the voices for. PMID- 11858594 TI - Unruffling the mystique of therapeutic touch. TT: the voices for. PMID- 11858595 TI - Unruffling the mystique of therapeutic touch. TT: the voices against. PMID- 11858596 TI - Unruffling the mystique of therapeutic touch. TT: the voices against. PMID- 11858597 TI - Unruffling the mystique of therapeutic touch. TT: the voices against. PMID- 11858598 TI - Unruffling the mystique of therapeutic touch. TT: the voices against. PMID- 11858599 TI - Unruffling the mystique of therapeutic touch. TT: the voices against. PMID- 11858600 TI - Stop dietary supplement health fraud. PMID- 11858601 TI - Shock wave-enhanced neovascularization at the tendon-bone junction: an experiment in dogs. AB - The purpose of the research was to study the phenomenon of neovascularization at the Achilles tendon-bone junction after low-energy shock wave application. The study was performed on eight mongrel dogs. The control specimens were obtained from the medial one-third of the right Achilles tendon-bone unit before shock wave application. Low-energy shock waves of 1000 impulses at 14 kV (equivalent to 0.18 mJ/mm2 energy flux density) were applied to the right Achilles bone-tendon junction. Biopsies were taken from the middle one-third of the Achilles tendon bone junction at 4 weeks and from the lateral one-third at 8 weeks, respectively, after shock wave application. The features of microscopic examination included the number of new capillaries and muscularized vessels, the presence and arrangements of myofibroblasts, and the changes in bone. New capillary and muscularized vessels were seen in the study specimens which were obtained in 4 weeks and in 8 weeks after shock wave application, but none were seen in the control specimens before shock wave application. There was a considerable geographic variation in the number of new vessels within the same specimen. Myofibroblasts were not seen in the control specimens. Myofibroblasts with haphazard appearance and intermediate orientation fibers were seen in all study specimens obtained at 4 weeks and predominantly intermediate orientation myofibroblast fibers at 8 weeks. There were no changes in bone matrix, osteocyte activity, and vascularization within the bone. Two pathologists reviewed each specimen and concurrence was achieved in all cases. The results of the study suggested that low-energy shock wave enhanced the phenomenon of neovascularization at the bone-tendon junction in dogs. PMID- 11858602 TI - ACFAS Universal Foot and Ankle Scoring System: First Metatarsophalangeal Joint and First Ray (module 1). AB - The American College of Foot and Ankle Surgeons presents the Universal Evaluation Scoring System to evaluate parameters related to foot and ankle surgery. This instrument was developed primarily to allow investigators a means of consistency in clinical assessments. The project was developed in four sections or modules. The first of these modules, First Metatarsophalangeal Joint and First Ray, is presented. This project is unique in that it is the first clinical scoring system of the foot and ankle to become validated by statistical analysis. PMID- 11858603 TI - Cortical bone pin fixation: a preliminary report on fixation of digital arthrodeses and distal chevron first metatarsal osteotomies. AB - From 1998 to 1999, the senior author utilized Allofix cortical bone pins for primary fixation of 40 first metatarsal distal chevron osteotomies and 30 digital arthrodeses in 42 patients. Radiographs of each patient were evaluated for the presence of graft incorporation, union of the osteotomy or fusion site, and postoperative migration of the osseous segments. In 70 procedures, the average time for graft incorporation as evaluated by the absence of graft visualization on radiographs was 4 months. Complications included one case of graft displacement. The results indicate that Allofix cortical bone pins are a reliable, safe, and effective method for the fixation of first metatarsal distal chevron osteotomies and phalangeal arthrodeses of the foot. PMID- 11858605 TI - Occult osteochondral fractures of the subtalar joint: a review of 10 patients. AB - Ten patients were treated who had occult osteochondral fractures of the subtalar joint that were not associated with dislocation. Nine osteochondral fractures involved the posterior facet. Seven patients had a stiff and painful subtalar joint, and an arthrodesis was performed. Three patients presented with adequate subtalar joint motion and were treated with physiotherapy. Detection of osteochondral fractures of the subtalar joint is difficult. The "early warning" signs are massive swelling without definitive bone injury on radiographic examination, and a failure to regain subtalar motion after a period of immobilization. Confirmatory evidence is obtained from CT scans or MRI. Initially, aggressive physiotherapy should be considered. Arthrodesis should be used for patients who remain symptomatic. PMID- 11858604 TI - Correction of complex foot deformities using the Ilizarov external fixator. AB - There are many drawbacks to using conventional approaches to the treatment of complex foot deformities, like the increased risk of neurovascular injury, soft tissue injury, and the shortening of the foot. An alternative approach that can eliminate these problems is the Ilizarov method. In the current study, a total of 23 deformed feet in 22 patients were treated using the Ilizarov method. The etiologic factors were burn contracture, poliomyelitis, neglected and relapsed clubfoot, trauma, gun shot injury, meningitis, and leg-length discrepancy (LLD). The average age of the patients was 18.2 (5-50) years. The mean duration of fixator application was 5.1 (2-14) months. We performed corrections without an osteotomy in nine feet and with an osteotomy in 14 feet. Additional bony corrective procedures included three tibial and one femoral osteotomies for lengthening and deformity correction, and one tibiotalar arthrodesis in five separate extremities. At the time of fixator removal, a plantigrade foot was achieved in 21 of the 23 feet by pressure mat analysis. Compared to preoperative status, gait was subjectively improved in all patients. Follow-up time from surgery averaged 25 months (13-38). Pin-tract problems were observed in all cases. Other complications were toe contractures in two feet, metatarsophalangeal subluxation from flexor tendon contractures in one foot, incomplete osteotomy in one foot, residual deformity in two feet, and recurrence of deformity in one foot. Our results indicate that the Ilizarov method is an effective alternative means of correcting complex foot deformities, especially in feet that previously have undergone surgery. PMID- 11858606 TI - Tibiofibular diastasis without ankle fracture: a review and report of two cases. AB - Diastasis of the distal tibiofibular syndesmosis is commonly seen with ankle fractures, but is a rare finding in the absence of fracture. The anatomy, biomechanics, mechanism of injury, evaluation, and radiographic assessment of the tibiofibular syndesmosis are reviewed. The authors present a review and two cases of traumatic ankle diastasis without ankle fracture. PMID- 11858607 TI - The posteromedial process fracture of the talus: a case report. AB - Injuries to the ankle tend to be some of the most common injuries seen in practice. Of these ankle injuries, fractures of the posteromedial process of the talus are often misdiagnosed as ankle sprains because of poor visualization on routine ankle radiographs. The examining physician must have a heightened awareness of this injury to accurately diagnose and treat this fracture. Proper treatment could help to lessen the possibility of post-traumatic arthritis to the ankle and subtalar joints. The authors present a case of a fracture of the posteromedial process of the talus, after a medial subtalar dislocation, treated with open reduction and internal fixation with bioabsorbable pins. PMID- 11858608 TI - Endoscopic gastrocnemius tenotomy. PMID- 11858609 TI - Strength comparison of allogenic bone screws, bioabsorbable screws, and stainless steel screw fixation. AB - Allogenic bone screws are new to the fixation market and have yet to be tested against current fixation materials. An in vitro comparison of the same sizes of stainless steel, bioabsorbable, and allogenic bone screws was undertaken to assess screw resistance to the forces of bending, pullout, and shear. Using aluminum plates to support the screws, forces up to 1000 Newtons were applied to six to eight samples of each type of screw. During each test, stainless steel screws withstood the maximum force that could be exerted by the testing apparatus without failing (bending, 113.9 +/- 11.8 N mean +/- SE; pullout 999.1 +/- 33.7 N; and shear, 997.5 +/- 108.8 N). In each test, compared to bioabsorbable screws, allogenic bone screws failed faster (pullout, allogenic: 12.4 +/- 1.1 seconds vs. bioabsorbable, 120.6 +/- 13.8 seconds; p = .001; bending, allogenic: 53.4 +/- 4.8 seconds vs. bioabsorbable, 201.9 +/- 11.1 seconds; p = .001; shear, allogenic 13.5 +/- 1.4 seconds vs. bioabsorbable, 43.8 +/- 0.9 seconds; p = .001) under equivalent (pullout: bioabsorbable, 385.0 +/- 18.4 N vs. allogenic, 401.0 +/- 35.9 N; p = .001) or lower (bending, allogenic: 4.7 +/- 0.2 N vs. bioabsorbable, 11.0 +/- 0.9 N; p = .675; shear, allogenic: 312.1 +/- 15.5 N vs. bioabsorbable 680.9 +/- 8.5 N; p = .001) loads, and in a highly variable fashion. Overall, the bioabsorbable screws withstood the forces of bending, pullout, and shear better than the allogenic screws, and stainless steel screws outperformed both bioabsorbable and allogenic screws. Despite these results, allogenic screws could still be useful in compliant patients who would benefit from their osteoconductive properties. PMID- 11858610 TI - HLA-DR expression in conjunctival cells after latanoprost. AB - The purpose of this randomized, controlled, masked study was to evaluate the effects of latanoprost 0.005% (Xalatan, Pharmacia and Upjohn, Milan, Italy) on the ocular surface in patients with ocular hypertension or primary open-angle glaucoma. The study group included 14 patients who underwent latanoprost 0.005% eye drops once a day in both eyes for 4 months. The control group underwent unpreserved substitute tears eye drops (Aquasalina, Bruschettini, Genoa, Italy) with the same posology and the same concentration of benzalkonium chloride (0.02%). Patients were controlled at 1, 3 and 4 months for symptoms, intraocular pressure, corneal sensitivity (Cochet-Bonnet test) and fluorescein staining. At the same control times, they underwent impression cytology and conjunctival brush cytology. Immunohistochemical techniques were used to detect HLA-DR expression in conjunctival cells. At each control, no significant statistical differences were found between the study and the control group, except for a higher expression of HLA-DR in the latanoprost group (p < 0.001). Since HLA-DR is a marker of ocular surface inflammation, this study suggests that a subclinical inflammatory reaction is present in the ocular surface of patients under latanoprost therapy. The pathway by which the inflammation is stimulated is yet unknown. PMID- 11858611 TI - Comparison of nitric oxide donors in lowering intraocular pressure in rabbits: role of cyclic GMP. AB - L-arginine-nitric oxide (NO) pathway participates in the physiology and in many pathological processes in the eye, such as glaucoma. The aim of the present study was to compare the ocular hypotensive effect of different NO-donors, and to get more information on the role of cyclic guanosine 3',5'-monophosphate (cGMP) in this process. The test compounds were administered topically or intravitreally in the eye of a normotensive rabbit. Intraocular pressure (IOP) was measured with a pneumatonometer after topical anesthesia. The metabolites of NO (nitrite, nitrate, NOx) and cGMP were assayed from the aqueous humor and plasma. NO synthase (NOS) protein expression was assayed in the ciliary body by Western blotting. The maximal lowering of IOP was achieved as follows: atriopeptin III (concentration 78 (microM, decrease in IOP 50%), atriopeptin II (84 (microM 37%). 8-Br-cGMP (90 mM, 37%), zaprinast + 8-Br-cGMP (1 mM + 90 mM, 34%), L-arginine (1 mM, 29%), SNP (40 mM, 28%), nitrosocaptopril (100 mM, 28%), S-nitrosothiol (SNOG) (10 mM, 27%), YC-1 (10 (microM, 25%), zaprinast + SNP (1 mM + 40 mM, 22%), spermine NONOate (100 mM, 20%) [corrected]. The decrease in IOP lasted for 2-5 hr, except with atriopeptin II and III, when IOP values were first normalized in 6 hr and 2 days, respectively. In conclusion, the results of the present study indicate that by increasing the activity of L-arginine/NO/cGMP-pathway it is possible to lower IOP in rabbits equally to the currently used antiglaucomatous drugs. PMID- 11858612 TI - Mechanism of vascular relaxation by cholinomimetic drugs with special reference to pilocarpine and arecoline. AB - The muscarinic receptor-mediated and non-muscarinic vascular effects of cholinomimetic drugs used in glaucoma were quantified. On the isolated rat aorta, the vascular tone induced by phenylephrine is functionally antagonized by cholinomimetic drugs. Based on EC50, the relative order of potency for the endothelium-dependent vascular relaxation was acetylcholine (0.05 microM) 1 > (+/ )-methacholine (0.35 microM) 1/7 > carbachol (0.63 microM) 1/12 > (+/-) aceclidine (1.26 microM) 1/25. The maximal effects of the four agonists varied between 82-87%. The muscarinic vascular relaxation of 0.03 microM to 100 microM pilocarpine was less than 15%. At high concentrations, pilocarpine had 1/20.000 the vascular activity of acetylcholine. Physostigmine failed to potentiate the vascular relaxation of exogenous acetylcholine, indicating the absence of acetylcholine esterase in the tissue. Arecoline, with an EC50 of 7.76 microM, was partly sensitive to the removal of the endothelium. Atropine treatment did not block the vascular effect of high concentrations of pilocarpine. Atropine, as expected, blocked the vascular effects of carbachol with K(B) = 3.2 nM. Pilocarpine produces vascular relaxation by its competition with spasmogens like phenylephrine, oxymetazoline, vasopressin or latanoprost. Arecoline also shares these properties with pilocarpine in the blood vessel. The molecular mechanism of the vascular effects as well as ocular clinical implications of cholinomimetic drugs is discussed. PMID- 11858613 TI - 3Alpha,5beta-tetrahydrocortisol effect on outflow facility. AB - 3Alpha,5beta-Tetrahydrocortisol (THF) was administered topically and intracamerally to ocular normotensive cynomolgus monkeys to determine whether it affects outflow facility. Monkeys received THF either topically at a dose of 2 x 5 microl drops of 300 microg/10 microl twice daily for 4 days (n = 4) or 3 times daily for 10 days (n = 4) with 10% DMSO as vehicle to the control eye, or intracamerally via 2 ml anterior chamber (AC) exchange of 30 microg/ml THF with vehicle, 0.1% DMSO, to the control eye followed by a second AC exchange using 300 microg/ml THF with vehicle to the control eye. Outflow facility was measured by a two-level constant pressure AC perfusion after administration of eye drops or after baseline outflow facility measurement and AC exchange with THF solution. The results showed no effect on outflow facility in normotensive cynomolgus monkeys. PMID- 11858614 TI - Effects of topical natural ergot alkaloids on intraocular pressure and aqueous humor dynamics in ocular normotensive rabbits. AB - Although it has been suggested that ergot derivatives may play a role in antiglaucoma therapy, little attention has been paid to the ocular hypotensive action of these drugs. Having previously reported in our laboratory that topical dihydroergocristine decreases intraocular pressure both in ocular normotensive and alpha-chymotrypsin-induced ocular hypertensive rabbits, the aim of the present work was to assess the effect of natural ergot alkaloids, ergocristine, alpha-ergocryptine and ergocornine, on the intraocular pressure and aqueous humor dynamics in ocular normotensive rabbits in order to further explore the ocular actions of these compounds. Intraocular pressure was measured with a pneumatonometer manometrically calibrated for the rabbit eye. Changes in tonographic facility of aqueous humor outflow and rate of aqueous humor inflow were evaluated in anesthetized rabbits. Natural ergot alkaloids were found to reduce intraocular pressure in ocular normotensive eyes in a dose-related fashion. These compounds decreased both tonographic outflow facility and, to a greater extent, aqueous humor inflow, which explains their final hypotensive effect. PMID- 11858615 TI - Effects of multiple dosing of epinephrine on aqueous humor dynamics in human eyes. AB - Numerous studies have provided conflicting evidence to explain the ocular hypotensive mechanism of action of epinephrine. Although epinephrine has been shown consistently to increase outflow facility, its effects on aqueous flow and uveoscleral outflow are not as clear. The purpose of this study was to clarify the effects of multiple doses of topical epinephrine on aqueous humor dynamics in human eyes. This was done by evaluating the four main parameters that determine steady state intraocular pressure. These parameters were assessed at baseline and after a week of twice-daily treatment of epinephrine hydrochloride 2% to one eye. Twenty-six human volunteers were enrolled in the study. Intraocular pressure was measured by pneumatonometry, aqueous flow and trabecular outflow facility by fluorophotometry, episcleral venous pressure by venomanometry and uveoscleral outflow by mathematical calculation. In epinephrine-treated eyes compared to baseline, intraocular pressure and aqueous flow were reduced from 21.2 +/- 0.3 to 17.1 +/- 0.2 mmHg (19%, p = .01) and 3.3 +/- 0.2 to 2.9 +/- 0.2 microl/min (12%, p = .03), respectively. Trabecular outflow facility obtained by fluorophotometry was increased from 0.18 +/- 0.02 to 0.26 +/- 0.03 microl/min/mmHg (44%, p = .02). Topical epinephrine did not significantly affect uveoscleral outflow or episcleral venous pressure. In conclusion, multiple topical doses of epinephrine lowered intraocular pressure in human volunteers by reducing aqueous humor formation and increasing trabecular outflow facility. The increase in uveoscleral outflow suggested by other studies was not observed. PMID- 11858616 TI - Alginic acid effect on carteolol ocular pharmacokinetics in the pigmented rabbit. AB - The effect of alginic acid addition to 1% or 2% carteolol solutions on the ocular penetration of the drug has been evaluated in the pigmented rabbit. During single dose studies, an increase in bioavailability ranging from 40% to 60% was observed in the aqueous humor and in the iris-ciliary body. During repeated dose studies, this increased ocular bioavailability of carteolol in the presence of alginic acid led to an equivalent concentration in the target tissue, although the dosage was only once a day compared with twice a day for the usual carteolol eyedrops. 14C-carteolol distribution studies demonstrated the binding of carteolol in pigmented ocular tissues. Thus, the presence of alginic acid as a new excipient supports a possible decrease in dosage regimen, while retaining sufficient ocular bioavailability to lower intraocular pressure. PMID- 11858617 TI - Pharmacological profile of oxaprozin eye drops. AB - The effects of eye drops containing a propionic acid derivative (oxaprozin) at 0.1% concentration on ocular inflammation produced by sodium arachidonate in the rabbit's eye were evaluated. Furthermore, the aqueous bioavailability of the drug formulation in the uninflamed and inflamed eyes was evaluated. Oxaprozin eye drops significantly reduced the signs of ocular inflammation elicited by sodium arachidonate on conjunctiva and iris. Oxaprozin treatment significantly reduced the levels of polymorphonuclear leukocytes and protein concentration in aqueous samples obtained from the eyes treated with arachidonate. Present data suggest, for the first time, that oxaprozin may be employed topically to prevent ocular reactions where the arachidonic acid cascade is activated. PMID- 11858618 TI - Distribution of cyclosporin A in the cornea after topical or oral administration. AB - The purpose of this study was to determine levels of cyclosporin A (CsA) in ocular tissues and fluids after oral or topical administration of CsA in humans requiring immunosuppression for penetrating keratoplasty and to evaluate the sensitivity of a high performance liquid chromatography coupled to electrospray mass spectrometry (LC-MS). High-risk penetrating keratoplasty was performed on 11 patients requiring 5 mg/kg of oral cyclosporin A and 9 patients with topical 0.5% (suspension, equivalent to 5 mg/ml)) preoperative immune suppression. Venous blood samples, as well as aqueous samples, were obtained for some patients, and the central 6.0-8.5 mm corneal buttons were removed and dissected into cornea epithelium, stroma and endothelium and analyzed for CsA levels by LC-MS. The mean values (concentration) of the specimens analyzed (obtained from 11 patients on oral CsA) showed 2.45 +/- 2.12 and 0.35 +/- 0.49 ng CsA/mg tissue in the epithelium and endothelium, respectively. A much smaller concentration, 0.13 +/- 0.14 ng CsA/mg tissue was observed in the stroma. There was a fair amount of CsA in the serum at the time of surgery but no detectable CsA in all aqueous humor samples analyzed. In cases of topical application, the mean concentration of CsA was similarly highest in the epithelium, 64.29 +/- 39.92 ng CsA/mg tissue, followed by the endothelium, 9.98 +/- 6.55 ng CsA/mg tissue, with the lowest levels found in the stroma. Topical application of CsA is able to achieve high levels of CsA in the corneal epithelium, higher than that achieved from oral administration of CsA. PMID- 11858619 TI - Corneal toxicity of intraocular hyaluronidase. AB - The purpose of this study was to examine the corneal toxicity of different preparations of intraocular hyaluronidase. SDS-PAGE analysis of bovine testicular hyaluronidase (Wydase) and chromatographically purified hyaluronidase (Sigma) was performed. These two preparations were injected into the anterior chamber of rabbits in amounts ranging from 1.5-150 IU (Wydase) and 1.5-300 IU (Sigma). A third set of rabbit eyes received Wydase vehicle alone or in combination with Sigma hyaluronidase. Treated control eyes were injected with saline. Slit lamp examination and indirect ophthalmoscopy were performed preoperatively and on postoperative days 1 and 7. Light microscopy of the corneas was performed. SDS PAGE of Wydase revealed numerous protein impurities, while Sigma demonstrated one protein band consistent with mammalian hyaluronidase. Persistent corneal edema, severe anterior chamber fibrin, and endothelial necrosis, were seen in the majority of eyes injected with Wydase in amounts of 50 IU and greater (n = 11). Thirty percent (30%) of the eyes injected with the Sigma preparation (n = 11) had localized corneal opacity similar to 50% of eyes injected with saline (n = 2). Of the rabbit eyes injected with the Wydase vehicle (n = 19), 68% had toxic changes. Intracameral injection of Wydase is toxic to the rabbit cornea in amounts of 50 IU and greater. A chromatographically purified preparation showed only transient local toxicity. Toxicity of Wydase may be due to protein impurities and the thimerosal-containing vehicle. PMID- 11858620 TI - HHS awards $69 million in grants to ensure early HIV care. PMID- 11858622 TI - Removal of treble fishhooks. PMID- 11858621 TI - Use of mindfulness meditation for fibromyalgia. PMID- 11858623 TI - Subclinical hyperthyroidism detected by screening: look before you treat. PMID- 11858624 TI - Promoting and prescribing exercise for the elderly. AB - Regular exercise provides a myriad of health benefits in older adults, including improvements in blood pressure, diabetes, lipid profile, osteoarthritis, osteoporosis, and neurocognitive function. Regular physical activity is also associated with decreased mortality and age-related morbidity in older adults. Despite this, up to 75 percent of older Americans are insufficiently active to achieve these health benefits. Few contraindications to exercise exist and almost all older persons can benefit from additional physical activity. The exercise prescription consists of three components: aerobic exercise, strength training, and balance and flexibility. Physicians play a key role in motivating older patients and advising them regarding their physical limitations and/or comorbidities. Motivating patients to begin exercise is best achieved by focusing on individual patient goals, concerns, and barriers to exercise. Strategies include the "stages of change" model, individualized behavioral therapy, and an active lifestyle. To increase long-term compliance, the exercise prescription should be straightforward, fun, and geared toward a patient's individual health needs, beliefs, and goals. PMID- 11858625 TI - Information from your family doctor. Exercise for the elderly. PMID- 11858626 TI - Subclinical hyperthyroidism: controversies in management. AB - Subclinical hyperthyroidism is an increasingly recognized entity that is defined as a normal serum free thyroxine and free triiodothyronine levels with a thyroid stimulating hormone level suppressed below the normal range and usually undetectable. The thyroid-stimulating hormone value is typically measured in a third-generation assay capable of detecting approximately 0.01 microU per mL (0.01 mU per L). Subclinical hyperthyroidism may be a distinct clinical entity, related only in part to Graves' disease or multinodular goiter. Persons with subclinical hyperthyroidism usually do not present with the specific signs or symptoms associated with overt hyperthyroidism. A detailed clinical history should be obtained, a physical examination performed and thyroid function tests conducted as part of an assessment of patients for subclinical hyperthyroidism and to evaluate the possible deleterious effects of excess thyroid hormone on end organs (e.g., heart, bone). A reasonable treatment option for many patients is a therapeutic trial of low-dose antithyroid agents for approximately six to 12 months in an effort to induce a remission. Further research regarding the etiology, natural history, pathophysiology, and treatment of subclinical hyperthyroidism is warranted. PMID- 11858627 TI - Problem drinking and alcoholism: diagnosis and treatment. AB - Alcoholism is one of the most common psychiatric disorders with a prevalence of 8 to 14 percent. This heritable disease is frequently accompanied by other substance abuse disorders (particularly nicotine), anxiety and mood disorders, and antisocial personality disorder. Although associated with considerable morbidity and mortality, alcoholism often goes unrecognized in a clinical or primary health care setting. Several brief screening instruments are available to quickly identify problem drinking, often a pre-alcoholism condition. Problem drinking can be successfully treated with brief intervention by primary care physicians. Alcohol addiction is a lifelong disease with a relapsing, remitting course. Because of the potentially serious implications of the diagnosis, assessment for alcoholism should be detailed. Alcoholism is treated by a variety of psychosocial methods with or without newly developed pharmacotherapies that improve relapse rates. Screening for problem drinking and alcoholism needs to become an integral part of the routine health screening questionnaire for adolescents and all adults, particularly women of child-bearing age, because of the risk of fetal alcohol syndrome. PMID- 11858628 TI - Information from your family doctor. Problem drinking--how to recognize it. PMID- 11858629 TI - Quinolones: a comprehensive review. AB - With the recent introduction of agents such as gatifloxacin and moxifloxacin, the traditional gram-negative coverage of fluoroquinolones has been expanded to include specific gram-positive organisms. Clinical applications beyond genitourinary tract infections include upper and lower respiratory infections, gastrointestinal infections, gynecologic infections, sexually transmitted diseases, and some skin and soft tissue infections. Most quinolones have excellent oral bioavailability, with serum drug concentrations equivalent to intravenous administration. Quinolones have few adverse effects, most notably nausea, headache, dizziness, and confusion. Less common but more serious adverse events include prolongation of the corrected QT interval, phototoxicity, liver enzyme abnormalities, arthropathy, and cartilage and tendon abnormalities. The new fluoroquinolones are rarely first-line agents and should be employed judiciously. Inappropriate use of agents from this important class of antibiotics will likely worsen current problems with antibiotic resistance. Applications of fluoroquinolones in biologic warfare are also discussed. PMID- 11858630 TI - Photo quiz. Koebner streak. PMID- 11858631 TI - ACOG issues recommendations on assessment of risk factors for preterm birth. American College of Obstetricians and Gynecologists. PMID- 11858632 TI - Pocket guide for care of HIV/AIDS patients. PMID- 11858633 TI - QSAR in Vivo. Proceedings of an American Chemical Society symposium, August 2000. PMID- 11858634 TI - Enhancement of binary QSAR analysis by a GA-based variable selection method. AB - Binary quantitative structure-activity relationship (QSAR) is an approach for the analysis of high throughput screening (HTS) data by correlating structural properties of compounds with a "binary" expression of biological activity (1 = active and 0 = inactive) and calculating a probability distribution for active and inactive compounds in a training set. Successfully deriving a predictive binary or any QSAR model largely depends on the selection of a preferred set of molecular descriptors that can capture the chemico-biological interaction for a particular biological target. In this study, a genetic algorithm (GA) was applied as a variable selection method in binary QSAR analysis. This GA-based variable selection method was applied to the analysis of three diverse sets of compounds, estrogen receptor (ER) ligands, carbonic anhydrase II inhibitors, and monoamine oxidase (MAO) inhibitors. Out of a variable pool of 150 molecular descriptors, predictive binary QSAR models were obtained for all three sets of compounds within a reasonable number of GA generations. The results indicate that the GA is a very effective variable selection approach for binary QSAR analysis. PMID- 11858635 TI - Beware of q2! AB - Validation is a crucial aspect of any quantitative structure-activity relationship (QSAR) modeling. This paper examines one of the most popular validation criteria, leave-one-out cross-validated R2 (LOO q2). Often, a high value of this statistical characteristic (q2 > 0.5) is considered as a proof of the high predictive ability of the model. In this paper, we show that this assumption is generally incorrect. In the case of 3D QSAR, the lack of the correlation between the high LOO q2 and the high predictive ability of a QSAR model has been established earlier [Pharm. Acta Helv. 70 (1995) 149; J. Chemomet. 10(1996)95; J. Med. Chem. 41 (1998) 2553]. In this paper, we use two-dimensional (2D) molecular descriptors and k nearest neighbors (kNN) QSAR method for the analysis of several datasets. No correlation between the values of q2 for the training set and predictive ability for the test set was found for any of the datasets. Thus, the high value of LOO q2 appears to be the necessary but not the sufficient condition for the model to have a high predictive power. We argue that this is the general property of QSAR models developed using LOO cross-validation. We emphasize that the external validation is the only way to establish a reliable QSAR model. We formulate a set of criteria for evaluation of predictive ability of QSAR models. PMID- 11858636 TI - Using the theoretical linear energy solvation energy relationship to correlate and predict nasal pungency thresholds. AB - The theoretical linear solvation energy relationship (TLSER) has been used to correlate and characterize 44 nasal pungency threshold (NPT) values in man with parameters derived from semi-empirical molecular orbital theory. The resulting relationship provides good correlative (R2 > 0.92) and predictive (R2cy > 0.88) capability. In addition, the TLSER parameters are used as a molecular probe to attempt to understand the fundamental properties influencing nasal pungency. PMID- 11858637 TI - Consensus scoring for ligand/protein interactions. AB - Several different functions have been put forward for evaluating the energetics of ligand binding to proteins. Those employed in the DOCK, GOLD and FlexX docking programs have been especially widely used, particularly in connection with virtual high-throughput screening (vHTS) projects. Until recently, such evaluation functions were usually considered only in conjunction with the docking programs that relied on them. In such studies, the evaluation function in question actually fills two distinct roles: it serves as the objective function being optimized (fitness function), but is also the scoring function used to compare the candidate docking configurations generated by the program. We have used descriptions available in the open literature to create free-standing scoring functions based on those used in DOCK and GOLD, and have implemented the more recently formulated PMF [J. Med. Chem. 42 (1999) 791] scoring function as well. The performance of these functions was examined individually for each of several data sets for which both crystal structures and affinities are available, as was the performance of the FlexX scoring function. Various ways of combining individual scores into a consensus score (CScore) were also considered. The individual and consensus scores were also used to try to pick out configurations most similar to those found in crystal structures from among a set of candidate configurations produced by FlexX docking runs. We find that the reliability and interpretability of results can be improved by combining results from all four functions into a CScore. PMID- 11858638 TI - COMPARE: a web accessible tool for investigating mechanisms of cell growth inhibition. AB - For more than 10 years the National Cancer Institute (NCI) has tested compounds for their ability to inhibit the growth of human tumor cell lines in culture (NCI screen). Work of Ken Paull [J. Natl. Cancer Inst. 81 (1989) 1088] demonstrated that compounds with similar mechanism of cell growth inhibition show similar patterns of activity in the NCI screen. This observation was developed into an algorithm called COMPARE and has been successfully used to predict mechanisms for a wide variety of compounds. More recently, this method has been extended to associate patterns of cell growth inhibition by compounds with measurements of molecular entities (such as gene expression) in the cell lines in the NCI screen. The COMPARE method and associated data are freely available on the Developmental Therapeutics Program (DTP) web site (http://dtp.nci.nih.gov/). Examples of the use of COMPARE on these web pages will be explained and demonstrated. Published by Elsevier Science Inc. PMID- 11858639 TI - In silico ADME/Tox: the state of the art. AB - The field of computational (in silico) ADME/Tox (absorption, distribution, metabolism, excretion, and toxicity) is receiving increased attention due a better appreciation that these molecular properties should be considered earlier in the drug discovery process. This report briefly reviews selected papers presented at the 220th National Meeting of the American Chemical Society, Washington, DC, 20-24 August, 2000, and describes the types of ADME/Tox computational models presented, the results obtained, and relevant recent publications that coincide with the work reported. PMID- 11858640 TI - Protein-ligand recognition using spherical harmonic molecular surfaces: towards a fast and efficient filter for large virtual throughput screening. AB - Molecular surfaces are important because surface-shape complementarity is often a necessary condition in protein-ligand interactions and docking studies. We have previously described a fast and efficient method to obtain triangulated surface meshes by topologically mapping ellipsoids on molecular surfaces. In this paper, we present an extension of our work to spherical harmonic surfaces in order to approximate molecular surfaces of both ligands and receptor-cavities and to easily check the surface-shape complementarity. The method consists of (1) finding lobes and holes on both ligand and cavity surfaces using contour maps of radius functions with spherical harmonic expansions, (2) superposing the surfaces around a given binding site by minimizing the distance between their respective expansion coefficients. This docking procedure capabilities was demonstrated by application to 35 protein-ligand complexes of known crystal structures. The method can also be easily and efficiently used as a filter to detect in a large conformational sampling the possible conformations presenting good complementarity with the receptor site, and being, therefore, good candidates for further more elaborate docking studies. This "virtual screening" was demonstrated on the platelet thrombin receptor. PMID- 11858641 TI - Molecular modelling of the differential interaction between several non-steroidal anti-inflammatory drugs and human prostaglandin endoperoxide H synthase-2 (h-PGHS 2). AB - The prostaglandin endoperoxide H synthase-1 (PGHS-1) and prostaglandin endoperoxide H synthase-2 (PGHS-2) are the targets of non-steroidal anti inflammatory drugs (NSAIDs). The high degree of selectivity for inhibition of PGHS-2 shown by certain compounds appears to stem from two mechanisms (time dependent, time-independent inhibition) by which they interact with each isoform. Molecular models of the complexes between indomethacin, fenamates, 2 phenylpropionic acids and the selective cyclooxygenase-2 (COX-2) inhibitors, with the cyclooxygenase active site of human PGHS-2 have been built by combining homology modelling, conformational searching and automated docking techniques. The stability of the resulting complexes has been assessed by molecular dynamics simulations combined with extended linear response calculations. The results allow us to identify regions of biological significance consistent with both X ray crystallographic and kinetic results. The selective PGHS-2 inhibitors exploit the extra space of a side-pocket in the active site of PGHS-2 that is not found in PGHS-1. The results obtained point out a marked relationship between the experimental affinity and the electrostatic interaction energy alone for a series of NSAIDs. Analysis of the structural and the energetic data provides evidence supporting that network of hydrogen bonds between Tyr355, Glu524, Arg120 and Arg513 might be involved in mediating the binding of the time-dependent inhibitors of PGHS-2. PMID- 11858642 TI - Immunodiagnosis of experimental Parelaphostrongylus tenuis infection in elk. AB - Elk infected with the meningeal worm, Parelaphostrongylus tenuis (Protostrongylidae), do not consistently excrete larvae in feces, making the current method of diagnosing live animals using the Baermann fecal technique unreliable. Serological diagnosis could prove more useful in diagnosing field infected animals but depends on the identification and availability of good quality antigen. To mimic field infections, 2 elk were inoculated with 6 infective L3 larvae of P. tenuis, and another 2 with 20 L3 larvae. Fecal samples were examined for nematode larvae using the Baermann technique and serum samples taken were tested for anti-P. tenuis antibody with ELISAs by using the excretory secretory (ES) products of L3, and sonicated adult worms as antigens. One animal passed first-stage larvae in its feces 202 days postinoculation, but passed none thereafter. The remaining 3 inoculated animals did not pass larvae. In contrast to parasite detection, antibodies against larval ES products were detected in all animals starting from 14 to 28 days postinoculation and persisted until the termination of the experiment on day 243 in 2 animals that harbored adult worms. Antibodies against somatic antigens of the adult worm were not detected until day 56 but also persisted until the end of the experiment in the animals with adult worms. In 2 elk that had no adult worms at necropsy, anti-ES antibodies were detected transiently in both, while anti-adult worm antibodies were present transiently in one. These findings confirm the superiority of P. tenuis larval ES products over somatic adult worm antigens as serodiagnostic antigens, as previously observed in studies of infected white-tailed deer, and extend the application of the newly developed ELISA test in diagnosing and monitoring cervids experimentally infected with P. tenuis. PMID- 11858643 TI - Determination of serum organic acids in puppies with naturally acquired parvoviral enteritis. AB - The purpose of the present study was to investigate the acid-base status and the serum concentration of organic acids in puppies with naturally occurring canine parvoviral enteritis. Between July 1999 and July 2000, 25 client-owned puppies admitted to the St. Louis Animal Emergency Clinic South for treatment of enteritis caused by parvovirus infection were used in our study. Control blood samples were collected from 22 healthy puppies less than 9 months of age. Serum organic acid concentrations were quantitatively determined by HPLC. Puppies infected with parvovirus had significantly lower plasma concentrations of sodium, potassium, chloride, and bicarbonate than controls. Although serum L-lactate tended to increase in some puppies with canine parvoviral enteritis, our study demonstrated that most affected puppies developed only mild compensated metabolic acidosis. None of the affected puppies had an elevated serum D-lactate concentration at admission. PMID- 11858644 TI - Recombinant equine interleukin-1beta induces putative mediators of articular cartilage degradation in equine chondrocytes. AB - Interleukin-1 is considered a central mediator of cartilage loss in osteoarthritis in several species, however an equine recombinant form of this cytokine is not readily available for in vitro use in equine osteoarthritis research. Equine recombinant interleukin-1beta was cloned and expressed and its effects on the expression and activity of selected chondrocytic proteins implicated in cartilage matrix degradation were characterized. Reverse transcriptase polymerase chain reaction methods were used to amplify the entire coding region of the equine IL-1beta mRNA, which was cloned into an expression vector, expressed in E. coli, and purified using a Ni2+ chromatographic method. The effects of the recombinant peptide on chondrocyte gene expression were determined by Northern blotting using RNA from equine chondrocyte cultures hybridized to probes for matrix metalloproteinases (MMP 1, MMP 3, MMP 13), tissue inhibitor of matrix metalloproteinases 1 (TIMP 1) and cyclooxygenase 2 (COX 2). Effects on selected mediators of cartilage degradation (nitrite concentrations and MMP activity) were determined using conditioned medium from reIL-1beta treated equine cartilage explant cultures. A recombinant peptide of approximately 21 kd was obtained. Northern blotting analyses revealed a marked up-regulation of expression of all MMPs, TIMP 1, and COX 2 in mRNA from treated chondrocytes. Furthermore, cartilage explants exposed to reIL-1beta had augmented collagenase/gelatinase and stromelysin activities as well as increased concentration of nitrite in conditioned media. The development of a biologically active, species-specific IL-1beta provides a valuable tool in the study of osteoarthritis pathophysiology and its treatment in horses. PMID- 11858646 TI - Decreases in serum apolipoprotein B-100 and A-I concentrations in cows with milk fever and downer cows. AB - Milk fever occurring during the peripartum period has been suggested to be caused by fatty liver developed during the non-lactating stage because diseased cows have increased serum concentrations of non-esterified fatty acids (NEFA) and show hepatic lipidosis. In cows with fatty liver and related diseases such as ketosis, serum concentrations of apolipoprotein (apo) B-100 and apoA-I are decreased. The purpose of the present study was to examine whether apoB-100 and apoA-I concentrations are similarly decreased in cows with milk fever. Apolipoprotein concentrations were also measured in cows with downer syndrome, which has been suggested to be related, at least in part, to milk fever. Compared with healthy cows during early lactation, apoB-100 and apoA-I concentrations were decreased in cows with milk fever and also in downer cows. In cows with milk fever, the decreases in apoB-100 and apoA-I concentrations were associated with increased NEFA and decreased cholesterol and phospholipid concentrations. However, in downer cows, serum lipid concentration changes were not as distinct as in cows with milk fever. These results, coupled with previous findings on the decreases in apoB-100 and apoA-I concentrations of cows with fatty liver-related diseases, suggest that fatty liver is involved in the development of milk fever and partly in that of downer cow syndrome. PMID- 11858645 TI - Effect of substrate selection on indirect immunofluorescence testing of canine autoimmune subepidermal blistering diseases. AB - The detection by indirect immunofluorescence (IIF) of circulating antibodies in the serum of dogs with autoimmune subepidermal blistering diseases (AISBD) was regarded for a long time as an unrewarding tool. It was, however, demonstrated in humans that the sensitivity of IIF assays depended on the selection of the substrates used. The effects of substrate selection on IIF tests was thus studied by examining sera from 12 dogs with AISBD tested against 8 different substrates from 3 different normal dogs. Patients with AISBD suffered from bullous pemphigoid (n = 4 sera), mucous membrane pemphigoid (n = 4 sera), and epidermolysis bullosa acquisita (n = 4 sera). Substrates included canine tongue, canine lip, canine dorsal haired skin, and ventral haired skin. The same 4 substrates were also split with salt splitting technique (using 1 M sodium chloride), in order to cleave the basement membrane within the lamina lucida and to expose the targeted antigens. The strength of the specific fluorescence of each slide was scored after processing for IIF testing with anti-canine IgG polyclonal antibody. Other criteria, such as background fluorescence, easiness of the interpretation, and variations within a same substrate, were also assessed. Intact canine lip and canine salt-split lip demonstrated consistently stronger intensity of fluorescence and a better ease of interpretation. We concluded that the performance of IIF tests with such substrates was a reliable tool for the detection of circulating IgG autoantibodies of canine patients with AISBD. PMID- 11858647 TI - Cliniconeuropathologic findings of familial frontal lobe epilepsy in Shetland sheepdogs. AB - We examined an epileptic focus by electroencephalography (EEG) by using an international 10-20 electrode system in 11 Shetland sheep dogs affected with familial idiopathic epilepsy. We also performed an evaluation of the amino acids in the cerebrospinal fluid (CSF) and a pathologic examination of the brains of 8 dogs that died from status epilepticus. Continuous electroencephalography demonstrated that an epileptic focus was initially detected in the frontal lobe, particularly the internal area, and that paroxysmal foci developed diffusely in other lobes of affected dogs with recurrent convulsions. The EEG analyses indicated spike and sharp wave complexes, which were considered to be paroxysmal discharges. An increased value for glutamate or aspartate was found in the CSF of some epileptic dogs. Histologically, acute neuronal necrosis and astrocytosis were distributed predominantly in the cingulate cortex and internal area of frontal cortex, less frequently in other areas of the cerebrum. The results of this study suggest that, initially, the dogs have an epileptic focus in the frontal lobe, and that the focus extends gradually to other areas of the cerebrum. Based on the distribution of neuronal necrosis and astrocytosis, acute neuronal damage may be related to the superexcitation of neurons following epilepsy. PMID- 11858648 TI - Neurohormonal and metabolic effects of medetomidine compared with xylazine in beagle dogs. AB - This study was aimed to investigate and compare the effects of medetomidine and xylazine on the blood level of some stress-related neurohormonal and metabolic variables in clinically normal dogs, especially focusing on time and dose relations of the effects. A total of 9 beagle dogs were used for 9 groups, which were treated with physiological saline solution (control), 10, 20, 40, and 80 microg/kg medetomidine, and 1, 2, 4, and 8 mg/kg xylazine, intramuscularly. Blood samples were taken at 10 times during 24 h from a central venous catheter. Plasma norepinephrine, epinephrine, cortisol, glucose, insulin, glucagon, and nonesterified fatty acid concentrations were determined. Both medetomidine and xylazine similarly and dose-dependently inhibited norepinephrine release and lipolysis. Medetomidine suppressed epinephrine release dose-dependently with greater potency than xylazine. Xylazine also tended to decrease epinephrine levels dose-dependently. The cortisol and glucagon levels did not change significantly in any treatment group. Both drugs suppressed insulin secretion with similar potency. Both medetomidine and xylazine increased glucose levels. The hyperglycemic effect of medetomidine, in contrast with xylazine, was not dose dependent at the tested dosages. The results suggested that the effect of medetomidine on glucose metabolism may not be due only to alpha2-adrenoceptor mediated actions. PMID- 11858649 TI - Prostaglandin E2 and reactive oxygen metabolite damage in the cecum in a pony model of acute colitis. AB - The objective of this project was to determine early tissue biochemical events associated with increased colonic secretion during the acute stage of castor-oil induced colitis by measuring cecal mucosal and submucosal malondialdehyde (MDA) and prostaglandin E2 (PGE2), levels in ponies. Intestinal tissue (inflamed or healthy) samples were obtained from 4 age- and sex-matched Shetland ponies. Biochemical methods were used to determine MDA and PGE2 levels in intestinal tissue samples from inflamed and healthy equine intestine. Inflamed tissue MDA and PGE2 levels increased with time after castor oil challenge and correlated with granulocyte infiltration, as determined by myeloperoxidase levels in a companion study. Elevated intestinal tissue MDA levels suggest that lipid peroxidation could be attributed to reactive oxygen metabolites (ROM) released from stimulated, recruited, and resident granulocytes. Tissue levels of MDA and PGE2 suggest a role for granulocyte-derived mediators of intestinal inflammation in the massive secretory response in cases of acute equine colitis. Tissue MDA and PGE2 levels may be useful laboratory tools to quantify and characterize intestinal secretory inflammatory responses in acute inflammatory conditions in the equine colon. PMID- 11858650 TI - Benign esophageal stricture in the dog and cat: a retrospective study of 20 cases. AB - Twenty animals with benign esophageal strictures are presented. Most of the esophageal strictures were thought to be related to gastroesophageal reflux during ovariohysterectomy and were located at the distal portion of the thoracic esophagus (caudal to the base of the heart). For the dilation procedure, the endoscope tip or a balloon catheter was used and the outcome was generally considered to be good. The endoscope tip was an adequate instrument for dilation in some cases. PMID- 11858651 TI - Demonstration of bovine herpesvirus type 1 and Mannheimia haemolytica antigens in specimens stored for up to 22 months in buffered formalin. AB - Bovine herpesvirus type 1 (BHV-1) and Mannheimia haemolytica antigens were demonstrated in lung tissues that were stored in 10% neutral phosphate buffered formalin for 1 to 22 months using the immunoperoxidase method. There were no differences observed in terms of labelling intensity and distribution of M. haemolytica antigens between specimens stored for 1 and 22 months. The labeling intensity in sections from 2-cm thick specimens was comparable to those from 0.2 cm thick specimens. There was no difference observed between pronase-treated and untreated sections. However, for BHV-1, the labeling intensity in untreated sections was reduced in tissues that had been stored from 12 to 22 months. Sections from thin specimens stored in neutral buffered formalin for 22 months exhibited a stronger staining intensity than those from thick specimens. PMID- 11858652 TI - Antibody response to an autogenous vaccine and serologic profile for Streptococcus suis capsular type 1/2. AB - An autogenous vaccine was developed, using sonicated bacteria, with a strain of Streptococcus suis capsular type 1/2. The objectives of this study were to evaluate the antibody response following vaccination and to assess the changes in antibody levels in pigs from a herd showing clinical signs of S. suis capsular type 1/2 infection in 6- to 8-week-old pigs. An enzyme-linked immunosorbent assay using the vaccine antigen was standardized. Results from a preliminary study involving 2 control and 4 vaccinated 4-week-old pigs indicated that all vaccinated pigs produced antibodies against 2 proteins of 34 and 43 kDa, respectively, and, in 3 out of 4 vaccinated pigs, against the 117-kDa muramidase released protein. For the serologic profile, groups of 30 pigs from the infected herd were blood sampled at 2, 4, 6, 8, and 10 weeks of age. The lowest antibody level was observed between weeks 6 and 8, presumably corresponding to a decrease in maternal immunity. A marked increase was seen at 10 weeks of age, shortly after the onset of clinical signs in the herd. For the vaccination field trial, newly weaned, one-week-old piglets were divided into 2 groups of 200 piglets each (control and vaccinated); blood samples were collected from 36 piglets in each group at 2-week intervals for 12 weeks. A significant increase (P < 0.05) in antibody response was observed 4 weeks following vaccination and the level of antibodies stayed high until the end of the experiment. In the control group, the increase was only observed at 13 weeks of age, probably in response to a natural infection. The response to the vaccine varied considerably among pigs and was attributed, in part, to the levels of maternal antibodies at the time of vaccination. No outbreak of S. suis was observed in the control or vaccinated groups, so the protection conferred by the vaccine could not be evaluated. PMID- 11858653 TI - Tyropeptins A and B, new proteasome inhibitors produced by Kitasatospora sp. MK993-dF2. II. Structure determination and synthesis. AB - The structures of tyropeptins A and B, new proteasome inhibitors produced by Kitasatospora sp. MK993-dF2, were determined by analysis of various NMR experiments. The 1H and 13C NMR of tyropeptins were complicated due to the presence of an aldehyde group. Therefore, tyropeptins were converted to their alcohols by sodium borohydride. These alcohol derivatives gave assignable NMR spectra. The stereochemistry of tyropeptins were determined by analysis of acid hydrolysis products from tyropeptins, and further confirmed by the total synthesis. The structures of tyropeptins A and B were found to be isovaleryl-L tyrosyl-L-valyl-DL-tyrosinal and n-butyryl-L-tyrosyl-L-leucyl-DL-tyrosinal, respectively. PMID- 11858654 TI - Neuroprotectins A and B, bicyclohexapeptides protecting chick telencephalic neuronal cells from excitotoxicity. I. Fermentation, isolation, physico-chemical properties and biological activity. AB - Glutamate, an excitatory amino acid, is known to induce neurotoxicity in central nervous system under abnormal conditions such as ischemia, hypoglycemia, epilepsy, Huntington's chorea, Parkinson's disease and Alzheimer's disease. In our search for neuroprotective agents of microbial origin against excitatory neurotoxins, we have isolated two new bicyclohexapeptides, neuroprotectins A and B, together with a known compound complestatin, from the fermentation broth of Streptomyces sp. Q27107. Neuroprotectins protected primary cultured chick telencephalic neurons from glutamate- and kainate-induced excitotoxicities in a dose-dependant fashion. PMID- 11858655 TI - Neuroprotectins A and B, bicyclohexapeptides protecting chick telencephalic neurons from excitotoxicity. II. Structure determination. AB - In the course of our search for neuroprotective agents of microbial origin against kainate-induced neurotoxicity, we have succeeded in the isolation of two new bicyclohexapeptides, neuroprotectins A and B, together with a known compound, complestatin, from the fermentation broth of Streptomyces sp. Q27107. They are closely related in structure to complestatin and possess an oxindolylalanine moiety in place of the tryptophan residue present in complestatin. PMID- 11858656 TI - CJ-13,981 and CJ-13,982, new squalene synthase inhibitors. AB - Two new squalene synthase (SSase) inhibitors, CJ-13,981 (I) and CJ-13,982 (II), were isolated from the fermentation broth of an unidentified fungus CL 15036. They inhibited human liver microsomal SSase with IC50s of 2.8 and 1.1 microM, respectively, but showed no inhibitory activity against human brain protein farnesyltransferase (PFTase) at 100 microM. Based on FAB-MS and NMR analyses, the structures of I and II were determined to be 3-hydroxy-3,4-dicarboxy-15 hexadecenoic acid and 3-hydroxy-3,4-dicarboxyhexadecanoic acid, respectively. PMID- 11858657 TI - CJ-19,784, a new antifungal agent from a fungus, Acanthostigmella sp. AB - A new antifungal agent, CJ-19,784 (I), was isolated from the fermentation broth of a fungus, Acanthostigmella sp. CL12082. Based on spectroscopic analyses, structure of I was determined to be 3'-bromo-2',5-dihydroxy-3,7,8-trimethoxy flavone. Compound I inhibits the growth of pathogenic fungi, Candida albicans, Cryptococcus neoformans and Aspergillus fumigatus with IC50 values of 0.11, 20 and 0.54 microg/ml, respectively. PMID- 11858658 TI - Goadsporin, a chemical substance which promotes secondary metabolism and morphogenesis in streptomycetes. I. Purification and characterization. AB - Streptomycetes, which belong to the Gram-positive bacteria, produce secondary metabolites and sporulate. The timing of starting the secondary metabolite production and the sporulation depends on environmental conditions such as nitrogen and carbon sources. In order to obtain a tool for understanding the regulation mechanism, we carried out screening for chemical substances that induce secondary metabolism and sporulation in streptomycetes and found an active substance from the culture broth of Streptomyces sp. TP-A0584. This substance designated goadsporin promoted the formation of red pigment and sporulation at a concentration of 1 microM in Streptomyces lividans TK23 which does not produce the pigment under normal growth conditions. Goadsporin is an oligopeptide consisting of 19 amino acids with the molecular formula C72H97N19020S2. Sporulation and/or secondary metabolite production was induced in 36 streptomycetes strains among 42 strains tested. These results suggest that goadsporin acts on a common regulation pathway for sporulation and secondary metabolism in streptomycetes and can be a powerful tool to analyze the regulation mechanism. PMID- 11858659 TI - Goadsporin, a chemical substance which promotes secondary metabolism and Morphogenesis in streptomycetes. II. Structure determination. AB - The structure of goadsporin was determined by using spectroscopic techniques. NMR analysis revealed that goadsporin consists of 19 amino acids, two of which are dehydroalanines (Deala), and six of which are cyclized to oxazoles (Oxz) and thiazoles (Thz) by dehydrative cyclization and dehydrogenation from serine, threonine and cysteine. NMR analysis established seven partial structures, and their sequence was determined by CID-MS/MS. Negative mode FAB-MS/MS gave product ions arising from charge-remote fragmentation that allowed determination of the sequence of the amino acid components as AcNH-Ala-MeOxz-Val-Deala-MeOxz-Ile-Leu Thz-Ser-Gly-Gly-MeOxz-Leu-Deala-Oxz-Ala-Gly-Thz-Val-OH. The chiral amino acids were determined by the advanced Marfey's method to have L-configurations. PMID- 11858660 TI - Tripropeptins, novel antimicrobial agents produced by Lysobacter sp. I. Taxonomy, isolation and biological activities. AB - Peptide antibiotics tripropeptins A, B, C, D and Z were isolated from cultured cells and broth of Lysobacter sp. The differences among these components are in the lengths of the alkyl side chain. Tripropeptins are active against Gram positive bacteria including MRSA in vitro. Bactericidal activity of tripropeptin C disappeared in the simultaneous presence of chloramphenicol, a bacteriostatic agent. PMID- 11858661 TI - CJ-15,801, a novel antibiotic from a fungus, Seimatosporium sp. AB - A novel antibiotic, CJ-15,801 (I), was isolated from the fermentation broth of a fungus, Seimatosporium sp. CL28611. The structure was determined to be a pantothenic acid analog having an alpha,beta-unsaturated carboxylic acid moiety by spectroscopic analyses. The compound inhibits the growth of multi-drug resistant (MDR) Staphylococcus aureus strains with MIC ranging from 6.25 to 50 microg/ml. PMID- 11858662 TI - Biosynthesis of the thiazolylpeptide antibiotic GE2270. AB - The biosynthesis of the antibiotic GE2270 in the producing microorganism Planobispora rosea was investigated by adding labelled amino acid precursors. Efficient incorporation of glycine and serine was observed, leading to specific enrichments of selected positions of the thiazole, oxazoline and pyridine rings. Furthermore, efficient enrichment of the C-, N- and O-methyl groups was detected. These results indicate that GE2270 is made through a biosynthetic route similar to that determined for other thiazolylpeptides. At the same time, the result point to an efficient route for the conversion of glycine into serine and methyl equivalents in Planobispora rosea. PMID- 11858663 TI - Some biological and biochemical activities of resormycin, a novel herbicidal antibiotic. AB - Biological and biochemical activities of resormycin were studied using unicellular green algae, Selenastrum capricornutum (abbreviated as Selena.), as a test organism. Resormycin inhibited the growth in vitro of Selena. more strongly in the dark than in the light. A weaker but more photo-stable derivative, (+/-) 2,3-dihydro-resormycin, showed more long-lasting activity against Selena. in the light. Resormycin started killing Selena. only after exposure for 2 days and longer, even at high concentrations. Resormycin at concentrations near IC50 rapidly inhibited incorporation of 3H-leu, but not 3H-UR or 3H-TdR, into the TCA insoluble fraction of Selena. Herbicidal activity of resormycin was confirmed using some crops and weeds. PMID- 11858664 TI - Synthesis and antibacterial activity of novel 1beta-methyl carbapenems with cycloalkylamine moiety at the C-2 position. AB - Novel 1beta-methyl carbapenems with a cycloalkylamine moiety as a side chain were synthesized and their structure-activity relationships were studied. These carbapenems showed potent antibacterial activities against a wide range of Gram positive and Gram-negative bacteria, and moderate urinary recovery when administered intraperitoneally in mice. PMID- 11858665 TI - A new trichostatin derivative, trichostatin RK, from Streptomyces sp. RK98-A74. PMID- 11858666 TI - Structure of Sch 528647: a new antitumor antibiotic related to fumagillin. PMID- 11858667 TI - MF-EA-705alpha & MF-EA-705beta, new metabolites from microbial fermentation of a Streptomyces sp. PMID- 11858668 TI - Migrastatin, a novel 14-membered ring macrolide, inhibits anchorage-independent growth of human small cell lung carcinoma Ms-1 cells. PMID- 11858669 TI - Syntheses of 2-aryliminooxazolidine derivatives as trehalase inhibitors. PMID- 11858670 TI - NK30424A and B, novel inhibitors of lipopolysaccharide-induced tumour necrosis factor alpha production, produced by Streptomyces sp. NA30424. PMID- 11858671 TI - Tyropeptins A and B, new proteasome inhibitors produced by Kitasatospora sp. MK993-dF2. I. Taxonomy, isolation, physico-chemical properties and biological activities. AB - Tyropeptins A and B, new proteasome inhibitors, were isolated from the culture broth of Kitasatospora sp. MK993-dF2. They were purified using ethyl acetate extraction, silica gel column chromatography, Sephadex LH-20 column chromatography and HPLC. Tyropeptin A inhibited the chymotrypsin-like (ChT-L) and trypsin-like (T-L) activities of 20S proteasome with IC50 values of 0.1 microg/ml and 1.5 microg/ml respectively, but did not inhibit the peptidylglutamyl-peptide hydrolyzing (PGPH) activity of 20S proteasome at a concentration of 100 microg/ml. The inhibitory activities of tyropeptin A were about two times as strong as that of tyropeptin B. Taxonomy of the producing strain is also described. PMID- 11858672 TI - Intravenous haloperidol. PMID- 11858673 TI - Advances in pacemaker technology. PMID- 11858674 TI - Liberating patients from mechanical ventilation: a team approach. PMID- 11858675 TI - Caring for adults with cystic fibrosis. PMID- 11858676 TI - Use of nasal cannula versus face mask after extubation in patients after cardiothoracic surgery. AB - Continuous reevaluation of protocols for patients' care is necessary to maintain high quality and cost-effectiveness in today's healthcare environment. A study of patients who had cardiothoracic surgery proved that after early extubation, patients could safely be given oxygen via nasal cannula with maintenance of acceptable oxygen saturation. The positive outcomes of this study were shorter exposure to oxygen at higher concentrations, greater compliance by patients, greater comfort for patients, and cost savings for the institution. This project was the result of thoughtful consideration and a willingness to question a standard practice that had been in existence in this cardiothoracic program for 20 years. Both patients and the institution benefit when we question why we do things and thoroughly evaluate our daily practice. All practitioners should always look for ways to change and improve practice for the betterment of patients. PMID- 11858677 TI - Understanding chest radiographs of infants and children: the AIR systematic approach. AB - A plain radiograph of the chest is the most frequently requested radiological image in infants and children. Assessing chest radiographs of infants and children can be challenging because of the age-related differences in normal anatomic structures, the effect of inhalation and exhalation during the imaging, and the direction of rotation. An understanding of basic radiographic principles is necessary before beginning to interpret chest films of infants and children. Use of the AIR approach can help nurses begin to understand and assess chest radiographs of infants and children in an orderly and systematic manner. PMID- 11858678 TI - Continuous Airway pressure monitoring. PMID- 11858679 TI - Applying the synergy model: clinical strategies. PMID- 11858680 TI - Instilling medication into chest tubes. PMID- 11858681 TI - A pain in the act: musings on the meaning for critical care nurses of the pain management standards of the joint commission on accreditation of healthcare organizations. PMID- 11858682 TI - Precepting for skills competencies is ongoing. PMID- 11858683 TI - Controversy over administration of phenytoin. PMID- 11858684 TI - Low staffing levels leave little time for care. PMID- 11858685 TI - Debate over use of infusion pumps. PMID- 11858686 TI - Treating acute ischemic stroke with intravenous alteplase. PMID- 11858687 TI - Bloodless medicine and surgery for patients having cardiac surgery. AB - Changes in patients' care mandate that nursing care be amenable to change. With the advent of new technology, systems of care delivery are evolving. Blood transfusion practices that were once considered standards are being reexamined and redefined. Nurses, in their roles as patients' advocates and educators, must be cognizant of changes in practice and be prepared to deliver competent care. The effectiveness of techniques of bloodless medicine and surgery originally designed to serve the needs of Jehovah's Witnesses is accepted today, and other groups of patients are choosing this type of care. Furthermore, as blood conservation strategies are refined and perfected, they most likely will be used by a greater number of patients undergoing cardiac surgery. Specific guidelines have been developed for point-of-care service for patients undergoing bloodless cardiac surgery. Nursing care throughout the hospital stay, from preadmission to discharge, is tailored to meet these patients' emotional and physical needs. Quality comprehensive nursing care is required to make bloodless medicine and surgery programs successful. PMID- 11858688 TI - Compatibility of commonly used intravenous infusions in a pediatric intensive care unit. PMID- 11858689 TI - Self-scheduling and staff incentives: meeting patient care needs in a neonatal intensive care unit. AB - Nursing staff and leadership in a resource-intensive NICU identified an innovative process for covering the unit's scheduling needs. Early concerns about the feasibility of achieving self-scheduling with a large staff were unwarranted. The use of a unit-based committee and the support of the nurse manager allowed us to develop a process that met the needs of the staff members and maintained the staffing standards of the unit. Contributing to the success of the self scheduling is a mechanism for recognizing and rewarding staff members who adjust their work schedules to meet the needs of the unit. Satisfaction among staff members with self-scheduling is high, and new employees cite the opportunity for self-scheduling as a contributing factor in their decisions to work in the NICU. PMID- 11858690 TI - Feeding critically ill patients: current concepts. AB - Provision of nutritional support to critically ill patients can be challenging. Critical care nurses must be aware of which patients require specific nutritional support, when to initiate nutritional support, and by which route to provide nutritional support. Consultation with a dietitian or nutritional support service can help facilitate this process. The key points in addressing these questions are (1) the nutritional status of the patient or the length of time he or she has been without significant nutrient intake, (2) whether the patient has a hypermetabolic condition that warrants the early use of nutritional support, and (3) the function of the patient's gastrointestinal tract. What to feed depends on the physiological state of the patient. Adjusting the nutrient composition of the feeding solution may prevent metabolic complications and may improve the overall outcome for the patient. PMID- 11858691 TI - Safely caring for patients with a laryngeal mask airway. PMID- 11858693 TI - Nursing and informatics: implications for critical care practice. PMID- 11858692 TI - The mechanisms of the anticoagulant and antithrombotic medications used in the treatment of acute coronary syndrome (ACS). PMID- 11858694 TI - Drug cards are a quick and portable reference. PMID- 11858695 TI - Protein expression in Down syndrome brain. AB - Down syndrome (DS) is the most common chromosomal abnormality associated with early mental retardation and neurological abnormalities followed by precocious age dependent Alzheimer-type neurode generation later in life. Knowledge of the pathological mechanisms involved in DS is far from complete, but overexpression of genes residing in chromosome 21 was considered to be the central point for the DS phenotype. In this regard, beta amyloid precursor protein (APP), CuZn superoxide dismutase (SOD1) and S100beta have been implicated in causing apoptosis, a mechanism thought to be responsible for neuronal loss in DS, in one way or another. The gene dosage hypothesis has been challenged, however, and dysregulation of expression of genes located on other chromosomes has been described, which may well be secondary to chromosomal imbalance or a direct consequence of the disease process. The present review focuses on the protein expression profile in DS and we postulate that abnormalities in the coordinated expression, as well as interaction of proteins may be responsible for the neuropathology of DS. A series of candidate proteins are discussed that may be directly causing or reflecting the DS phenotype, in particular the brain abnormalities in DS. PMID- 11858696 TI - Proteomics: current technologies and applications in neurological disorders and toxicology. AB - Proteomics is the science that studies the proteins in general and in particular their changes, resulting from various disorders or the effect of external factors, such as toxic agents. It has as goal the detection of novel drug targets, diagnostic markers and the investigation of biological events. Proteomics has emerged the last few years and its major difference from the previously existing protein analytical techniques is that it does not analyze the proteins one by one, but in a possibly automated, large-scale mode. In this article, the state of the art of proteomics in our laboratory is presented, as well as selected applications of proteomics in the study of disorders of the central nervous system and of toxic events. PMID- 11858697 TI - Catabolism of methionine and threonine in vitro by mixed ruminal bacteria and protozoa. AB - In vitro studies were conducted to examine the metabolism of methionine (Met) and threonine (Thr) using mixed ruminal bacteria (B), mixed ruminal protozoa (P), and a combination of these two (BP). Rumen microorganisms were collected from fistulated goats fed with lucerne cubes (Medicago sativa) and a concentrate mixture twice a day. Microbial suspensions were anaerobically incubated with or without 1 mM each of the substrates at 39 degrees C for 12h. Met, Thr and their related amino compounds in both the supernatants and microbial hydrolyzates of the incubation were analyzed by HPLC. Met was degraded by 58.7, 22.1, and 67.3% as a whole in B, P, and BP suspensions, respectively, during 12h incubation. In the case of Thr, these values were 67.3, 33.4, and 76.2% in B, P, and BP, respectively. Met was catabolized by all of the three microbial suspensions to methionine sulfoxide and 2-aminobutyric acid. Catabolism of Thr by B and BP resulted in the production of glycine and 2-aminobutyric acid, while P produced only 2-aminobutyric acid. From these results, the existence of diverse catabolic routes of Met and Thr in rumen microorganisms was indicated. PMID- 11858698 TI - Conservation of the basic pattern of cellular amino acid composition of archaeobacteria during biological evolution and the putative amino acid composition of primitive life forms. AB - Previous studies showed that the cellular amino acid composition obtained by amino acid analysis of whole cells, differs such as eubacteria, protozoa, fungi and mammalian cells. These results suggest that the difference in the cellular amino acid composition reflects biological changes as the result of evolution. However, the basic pattern of cellular amino acid composition was relatively constant in all organisms examined. In the present study, we examined archaeobacteria, because they are considered important in understanding the relationship between biological evolution and cellular amino acid composition. The cellular amino acid compositions of Archaeoglobus fulgidus, Pyrococcus horikoshii, Methanobacterium thermoautotrophicum and Methanococcus jannaschii differed slightly from each other, but were similar to those determined from codon usage data, based on the complete genomes. Thus, the cellular amino acid composition reflects biological evolution. We suggest that primitive forms of life appearing on earth at the end of prebiotic evolution had a similar-cellular amino acid composition. PMID- 11858700 TI - Kinetics of taurine depletion and repletion in plasma, serum, whole blood and skeletal muscle in cats. AB - The relationship between taurine concentrations of plasma, whole blood, serum and skeletal muscle during taurine depletion and repletion was investigated in cats, to identify the most useful indicators of taurine status. Sixteen cats were fed a purified diet containing either 0 or 0.15 g/kg taurine for 5 months. Treatments were then reversed and the taurine concentration was measured during repletion and depletion phases. Plasma taurine exhibited the fastest rate (slow component) of depletion (t 1/2 = 4.8 wk), followed by serum (5.3 wk), whole blood (6.2 wk), and skeletal muscle (11.2 wk). Whole blood taurine was the first to replete at a rate of 0.74 wk to 1/2 maximal repletion, followed by serum (2.1 wk), skeletal muscle (3.5 wk), and plasma (3.5 wk). Whole blood more closely reflected skeletal muscle taurine concentrations than plasma during depletion, while plasma taurine concentrations appear to be the most valuable predictor of skeletal muscle taurine concentrations during repletion. This study suggests that the best clinical method to evaluate the taurine status of the cat is the determination and interpretation of both plasma and whole blood taurine concentrations. PMID- 11858701 TI - Saccharide induced racemization of amino acids in the course of the Maillard reaction. AB - The formation of D-amino acids on heating aqueous solutions of protein L-amino acids at pH 2.5 and pH 7.0 together with glucose, fructose or saccharose was investigated by enantioselective gas chromatography. The saccharide induced partial racemization (epimerisation) of L-amino acids is attributed to the Maillard reaction. PMID- 11858699 TI - The effect of high protein diet on urea and guanidino compound levels in renal insufficient mice. AB - Nephrectomy in mice provokes a decrease in creatinine clearance (CTN(Cl)) and an increase in urea and specific guanidino compound (GC) concentrations in blood and other tissues. Our purpose was to investigate the influence of high protein diet (HPD) on CTN(Cl), urea and GC levels in NX mice. Mice were nephrectomized or sham operated and subdivided in groups to study five diet conditions. At the end of each experiment, 10 days and 30 days postsurgery, urine and blood were collected for determination of urea and GCs, including creatinine. HPD resulted in significantly higher CTN(Cl) values in sham-operated mice than those observed in mice under normal protein diet, 10 days as well as 30 days postnephrectomy. HPD induced significant increases in plasma urea, guanidinosuccinic acid, argininic acid and a-keto-delta-guanidinovaleric acid concentration 10 days postsurgery but not 30 days postsurgery. HPD coincided with significantly higher excretion of urea, guanidinosuccinic acid, alpha-keto-delta-guanidinovaleric acid, creatine, argininic acid and gamma-guanidinobutyric acid in sham-operated and nephrectomized mice 10 days postsurgery. Our results show that HPD induces supplementary (to nephrectomy) increases of urea and GCs in the early postsurgery period but not in the later phase. PMID- 11858702 TI - Further ultrastructural characterization of the intranuclear ring-shaped bodies of the plant Lacandonia schismatica. AB - Ring-shaped bodies are found in the nucleus of Lacandonia schismatica, a rare plant with the sexual organs inverted. They are 0.5-microm-diameter structures that present an electron-dense external ring surrounding a central core. Ultrastructural studies indicate that these bodies contain RNA. The external ring is labeled with antibodies against small nuclear ribonucleoproteins, suggesting that they may be involved in pre-mRNA metabolism. In the present work we further characterized these intranuclear ring-shaped structures by serial-sectioning analysis. Moreover, we tested the presence of additional molecular elements related to pre-mRNA metabolism, such as SR proteins and poly(A)(+) RNA, using immunoelectron microscopy and ultrastructural in situ hybridization. Our results show that these nuclear bodies are spherical. They contain SR proteins involved in splicing and postsplicing events and little to no poly(A)(+) RNA. We also found similar nuclear bodies in other plant and animal species. Therefore, ring shaped bodies in L. schismatica are spherical, highly compartmentalized nuclear structures that may be involved in pre-mRNA metabolism. PMID- 11858703 TI - The formation of actin rods composed of actin tubules in Dictyostelium discoideum spores. AB - A new type of actin rod formed in both the nucleus and the cytoplasm, as well as tyrosine phosphorylation of actin, is implicated in the maintenance of dormancy and viability of Dictyostelium discoideum spores. Here the ultrastructure of the rods and their relationship to the phosphorylation of actin were examined. The rods first appeared in premature spores at the midculmination stage as bundles composed of actin tubules hexagonally cross-linked. The 13-nm-diameter bundles were composed of three actin filaments. Formation of the actin rods begins during the late culmination stage and proceeds until 2 days after completion of fruiting bodies. The physical events occur in the following order; association of several modules of bundles, close packing and decrease in diameter of actin tubules, elongation of rods across the nucleus or the cytoplasm. Actin phosphorylation levels increased at the late culmination stage and reached a maximum level 12 h later. Immediately following activation of spore germination, actin was rapidly dephosphorylated, followed shortly thereafter by the disappearance of rods. Shortened actin tubules once again became arranged in a hexagonal pattern. This hexagonal arrangement of actin tubules is possibly involved in rod formation and disappearance and does not depend upon actin phosphorylation. In contrast, rod maturation processes may correlate with actin phosphorylation. PMID- 11858704 TI - Correlation of topographic surface and volume data from three-dimensional electron microscopy. AB - Three-dimensional(3D) reconstructions from tilt series in an electron microscope show in general an anisotropic resolution due to an instrumentally limited tilt angle. As a consequence, the information in the z direction is blurred, thus making it difficult to detect the boundary of the reconstructed structures. In contrast, high-resolution topography data from microscopic surface techniques provide exactly complementary information. The combination of topographic surface and volume data leads to a better understanding of the 3D structure. The new correlation procedure presented determines both the height scaling of the topographic surface and the relative position of surface and volume data, thus allowing information to be combined. Experimental data for crystalline T4 bacteriophage polyheads were used to test the new method. Three-dimensional volume data were reconstructed from a negatively stained tilt series. Topographic data for both surfaces were obtained by surface relief reconstruction of electron micrographs of freeze-dried and unidirectionally metal-shadowed polyheads. The combined visualization of volume data with the scaled and aligned surface data shows that the correlation technique yields meaningful results. The reported correlation method may be applied to surface data obtained by any microscopic technique yielding topographic data. PMID- 11858705 TI - The effects of deimination of myelin basic protein on structures formed by its interaction with phosphoinositide-containing lipid monolayers. AB - The recombinant 18.5-kDa charge isoform of murine myelin basic protein (rmMBP) is unmodified posttranslationally and was used to study the effects of deimination, i.e., the conversion of arginyl to citrullinyl residues, on the protein's interactions with itself and with lipids. The unmodified species rmMBP-Cit(0) (i.e., containing no citrullinyl residues) interacted with binary monolayers containing acidic (phosphatidylinositol) and nickel-chelating lipids to form paracrystalline arrays with 4.8-nm spacing. A sample of protein was deiminated to an average of 9 citrullinyl residues per molecule of protein, yielding rmMBP Cit(9). Under both low- and high-salt conditions, this species formed better ordered domains than rmMBP-Cit(0), viz., planar crystalline assemblies. Thus, deimination of MBP resulted in a significant alteration of its lipid-organizing and self-interaction properties that might be operative in myelin in vivo, especially in progression of the autoimmune disease multiple sclerosis. Comparisons of amino acid sequences indicated significant similarities of MBP with filaggrin, a protein that is deiminated in another autoimmune disease, rheumatoid arthritis, suggesting that comparable epitopes could be targeted in both pathologies. In contrast, binary lipid monolayers consisting of phosphatidylinositol-4-phosphate (or phosphatidylinositol-4,5-bisphosphate) and a nickel-chelating lipid formed helical tubular vesicular structures, which appeared to be induced and/or stabilized by rmMBP, especially in its deiminated form. Sequence comparisons with other actin- and phosphoinositide-binding proteins (vinculin, ActA, MARCKS) suggested that the carboxyl-terminal segment of MBP could form an amphipathic alpha helix and was the phosphoinositide binding site. PMID- 11858706 TI - Regional structural and viscoelastic properties of fibrocartilage upon dynamic nanoindentation of the articular condyle. AB - Fibrocartilage,a tissue with macromaterial properties between dense fibrous tissue and hyaline cartilage, is not well understood in its ultrastructure and regional viscoelastic properties. Here nanoindentation with atomic force microscopy was performed on fresh fibrocartilage samples of rabbit jaw joint condyles. Each sample was divided into anteromedial, anterolateral, posteromedial, and posterolateral regions for probing and topographic imaging in 2 x 2 microm and 10 x 10 microm scan sizes. Young's moduli differed significantly among these regions in a descending gradient from the anteromedial (2.34 +/- 0.26 MPa) to the posterolateral (0.95 +/- 0.06 MPa). The Poisson ratio, defined as lateral strain over axial strain, had the same gradient distribution: highest for the anteromedial region (0.46 +/- 0.05) and lowest for the posterolateral region (0.31 +/- 0.05). The same four regions showed a descending gradient of surface roughness: highest for the anteromedial (321.6 +/- 13.8 nm) and lowest for the posterolateral (155.6 +/- 12.6 nm). Thus, the regional ultrastructural and viscoelastic properties of fibrocartilage appear to be coregulated. Based on these region-specific gradient distributions, fibrocartilage is constructed to withstand tissue-borne shear stresses, which likely propagate across its different regions. A model of shear gradient and concentric gradient is proposed to describe the region-specific capacity of fibrocartilage to sustain shear stresses in tendons, ligaments, joints, and the healing bone across species. PMID- 11858707 TI - Polymer-mediated compaction and internal dynamics of isolated Escherichia coli nucleoids. AB - Nucleoids of Escherichia coli were isolated by osmotic shock under conditions of low salt in the absence of added polyamines or Mg(2+). As determined by fluorescence microscopy, the isolated nucleoids in 0.2 M NaCl are expanded structures with an estimated volume of about 27 microm(3) according to a procedure based on a 50% threshold for the fluorescence intensity. The nucleoid volume is measured as a function of the concentration of added polyethylene glycol. The collapse is a continuous process, so that a coil-globule transition is not witnessed. The Helmholtz free energy of the nucleoids is determined via the depletion interaction between the DNA helix and the polyethylene glycol chains. The resulting compaction relation is discussed in terms of the current theory of branched DNA supercoils and it is concluded that the in vitro nucleoid is crosslinked in a physical sense. Despite the congested and crosslinked state of the nucleoid, the relaxation rate of its superhelical segments, as monitored by dynamic light scattering, turns out to be purely diffusional. At small scales, the nucleoid behaves as a fluid. PMID- 11858708 TI - The organization of mature Rous sarcoma virus as studied by cryoelectron microscopy. AB - We have studied the organization of mature infectious Rous sarcoma virus (RSV), suspended in vitreous ice, using transmission electron microscopy. The enveloped virions are spherical in shape, have a mean diameter of 127 nm, and vary significantly in size. Image processing reveals the presence of the viral matrix protein underlying the lipid bilayer and the viral envelope proteins external to the lipid bilayer. In the interior of the virus, the characteristic mature retroviral core is clearly imaged. In contrast to lentiviruses, such as human immunodeficiency virus, the core of RSV is essentially isometric. The capsid, or external shell of the core, has a faceted, almost polygonal appearance in electron micrographs, but many capsids also exhibit continuous surface curvature. Cores are not uniform in size or shape. Serrations observed along the projected faces of the core suggest a repetitive molecular structure. Some isolated cores were observed in the sample, confirming that cores are at least transiently stable in the absence of the viral envelope. Using an approach grounded in geometric probability, we estimate the size of the viral core from the projection data. We show that the size of the core is not tightly controlled and that core size and virion size are positively correlated. From estimates of RNA packing density we conclude that either the RNA within the core is loosely packed or, more probably, that it does not fill the core. PMID- 11858709 TI - Lipid-layer crystallization and preliminary three-dimensional structural analysis of AcrA, the periplasmic component of a bacterial multidrug efflux pump. AB - The multidrug efflux complex AcrAB-TolC confers intrinsic drug resistance in Escherichia coli by pumping antibiotics out of the cell. We determined a low resolution (20 A) structure of AcrA, the periplasmic component, by electron crystallography. Expressed with a His-tag at its carboxyl-terminus, the protein bound to lipid layers containing the nickel-chelating phospholipid DOGS-NTA. Under the lipid layers, AcrA crystallized in layer group P2(1)22, with a unit cell size of 157 by 95 A and a thickness of about 100 A. The four asymmetric units in the unit cell are organized into what appears to be two rings, each with a central opening of 30 A in diameter. Within each ring, the density can be interpreted as following a pseudo-helical path, approximately 210 A long. This length matches that of monomeric AcrA in solution, previously estimated by light scattering and hydrodynamic measurements. On one side the density has a tubular shape, with a thickness of about 25 A, while on the other side the densities of the upper and lower parts of the pseudo-helical path are fused into a shield. PMID- 11858710 TI - Methods for preparation of recombinant cytokine proteins V. mutant analogues of human interferon-gamma with higher stability and activity. AB - Mutant analogues of recombinant human immune interferon (IFN-gamma) with higher stability and biological activity were prepared. Depending on the analogue, protein structure modification might involve introduction of an intramonomer disulfide bond (through replacements of Glu7Cys and Ser69Cys), C-terminal shortening by 10 amino acid residues, as well as Gln133Leu substitution in truncated variant. Isolation, purification, and renaturation of the IFN-gamma analogues expressed in Escherichia coli as inclusion bodies were performed according to the scheme developed earlier for wild-type protein. The main idea of this scheme is to remove cellular impurities before recombinant protein renaturation. Folding kinetics of IFN-gamma was studied by reversed-phase HPLC. IFN-gamma and mutant proteins were characterized by their thermal stability and biological activity. Introduction of the intramolecular disulfide bond together with C-terminal shortening and replacement of C-terminal residue was shown to result in increasing the thermal stability by 19 degrees C and four times enhancement of biological activity compared with intact IFN-gamma molecule. PMID- 11858711 TI - Separation of an isoenzyme of polyphenol oxidase from Duranta plumieri by expanded bed chromatography. AB - Aqueous extracts of seeds of Duranta plumieri were found to be rich in polyphenol oxidase activity. The anion-exchange chromatography of the crude extract on Streamline DEAE resolved the activity into three fractions. The major fraction (77% of the total activity) was further purified by treating it with concanavalin A-agarose in the batch mode. The enzyme preparation eluting with alpha methylmannoside showed a single band on SDS-PAGE. The minimum molecular weight corresponded to 14,000 Da. The K(m) and V(max) of this isoenzyme were found to be 7.1 mM and 73.5 U ml(-1) min(-1) respectively. The k(cat) of this isoenzyme was calculated to be 8235 s(-1). The isoenzyme also showed the phenomenon of latency and the activity could be enhanced by 196% on heating it at 55 degrees C for 30 min. PMID- 11858712 TI - Purification and characterization of three immunodominant proteins (38, 30, and 16 kDa) of Mycobacterium tuberculosis. AB - Specific mycobacterial antigens are an important prerequisite in the serodiagnosis of tuberculosis. Many studies have reported the use of both native and recombinant proteins. Even though recombinant proteins can form standardized reagents with unlimited supply, their diagnostic test characteristics were not satisfactory in some cases. In this study we have purified the 38-, 30- (antigen 85B), and 16-kDa native antigens of Mycobacterium tuberculosis by procedures with limited number of steps. Starting with the secreted antigens of M. tuberculosis H37Rv, the 38-kDa form was purified by preparative isoelectric focusing, followed by preparative electrophoresis. Separation of antigen 85 components was achieved by anion-exchange chromatography, followed by hydrophobic interaction chromatography. Gel-permeation chromatography was employed for the isolation of the 16-kDa form, from the cytosol fraction of M. tuberculosis H37Rv. By using a minimal number of steps, considerable yields of these proteins were obtained without loss of immunological activity. The native proteins purified were characterized by analytical two-dimensional electrophoresis, HPLC, and circular dichroism studies. Conformation of the native 38-kDa form purified in our laboratory was different from that of the recombinant 38-kDa form from the WHO Bank. The identities of these native antigens were established by immunoblotting with known monoclonal antibodies from the WHO Bank. PMID- 11858713 TI - Characterization of a truncated soluble form of the baculovirus (AcMNPV) major envelope protein Gp64. AB - A truncated tagged form of the Autographica californica multiple nuclear polyhedrosis virus major surface glycoprotein, gp64, has been expressed using the baculovirus expression system and purified to homogeneity by immune-affinity chromatography. The protein, which is responsible for virus-cell fusion, was a trimer in solution and retained this oligomeric form at pH 5, the pH of fusion. Circular dichroism spectroscopy indicated a protein with mixed alpha-helix and beta-sheet content that did not undergo significant change at pH 5. The soluble protein showed no detectable binding to the insect cell surface. These data suggest a novel fusion mechanism for gp64 compared to models such as the influenza HA. In a crystal screen, deglycosylated, but not glycosylated, preparations of the protein were found to form small needle-shaped crystals that may form the basis of a dedicated structural study. PMID- 11858714 TI - Expression, characterization, and biochemical properties of recombinant human salivary amylase. AB - Human salivary amylase, a major component of human salivary secretions, possesses multiple functions in the oral cavity. It is the only enzyme in saliva capable of degrading oligosaccharides, which are used by the oral microflora for nutritional purposes. In order to understand its role in disease processes such as caries, we have undertaken the structure-function analyses of amylase. In this regard, the nonglycosylated human salivary amylase was expressed in a baculovirus expression system. The native and the recombinant amylases exhibit similar biochemical as well as biophysical properties. Unlike recombinant human pancreatic amylase, recombinant human salivary amylase is not glycosylated when expressed in a baculovirus system as determined from the crystal structure determination of the recombinant enzyme. Therefore, this system is suitable for further structure function work without resorting to enzymatic removal of the carbohydrate chain. Details of the expression, purification, and biophysical properties will be presented. PMID- 11858715 TI - Optimizing functional versus total expression of the human mu-opioid receptor in Pichia pastoris. AB - The expression of the EGFP-human mu-opioid receptor fusion protein in the methylotrophic yeast Pichia pastoris was optimized and monitored using both fluorescence and ligand-binding experiments. A set of parameters, including gene copy number, strain type, temperature, pH, and methanol inducer levels, was studied for its effect on the production of the recombinant protein. We show here that the expression level is optimal after 10 h of promoter induction and that the maximum is reached at a lower temperature and a higher pH than normally used. The optimized conditions have allowed a fourfold increase of the ligand-binding active form of the receptor, whereas the total expression level determined by EGFP fluorescence measurements was not modified. PMID- 11858716 TI - Characterization of a Brassica napus myrosinase expressed and secreted by Pichia pastoris. AB - In Brassica napus three different gene families with different temporal and tissue-specific expression and distribution patterns encode myrosinases (thioglucoside glucohydrolases, EC 3.2.3.1). Myrosinases encoded by the MA gene family are found as free and soluble dimers, while myrosinases encoded by the MB and MC gene families are mainly found in large insoluble complexes associated with myrosinase-binding proteins and myrosinase-associated proteins. These large complexes impede purification and characterization of MB and MC myrosinases from the plant. We used Pichia pastoris to express and secrete functional recombinant MYR1 myrosinase from B. napus to allow further characterization of myrosinase belonging to the MB gene family. The purified recombinant myrosinase hydrolyzes sinigrin with a K(m) of 1.0 mM; the specific activity and calculated k(cat)/K(m) were 175 U/mg and 1.9 x 10(5) s(-1) M(-1), respectively. A novel in-gel staining method for myrosinase activity is presented. PMID- 11858717 TI - Production, purification, and characterization of recombinant prohormone convertase 5 from baculovirus-infected insect cells. AB - The discovery of the prohormone convertase (PC) family of enzymes has provided several good candidates (PC1, PC2, and PC5) for the enzymes responsible for the endoproteolytic cleavage of procholecystokinin (pro-CCK). Determination of the role of individual pro-hormone convertases in the processing of pro-CCK is complicated because several of these enzymes are found in endocrine tumor cells expressing CCK mRNA and in identified neurons in the brain. Production of active recombinant PC5 permits the determination of its ability to cleave substrates related to pro-CCK. Active PC5, secreted from baculovirus-infected Sf9 cells, was partially purified by ion-exchange chromatography. Western blot analysis confirmed the presence of the active form of the enzyme in infected cell media and its absence from uninfected cell media. The enzyme is most active at acidic pH 6.5 and is maximally activated by 5 mM calcium. PC5 was able to cleave both monobasic and dibasic substrates without a requirement for a basic residue at P-4 and it displayed a K(m) in the micromolar range. The enzyme was inhibited by EDTA, 1,10-phenanthroline, and p-CMS, as well as by two specific PC inhibitors. This is the first reported preparation of active recombinant PC5. Like the other members of its family, it has the correct catalytic characteristics in vitro to play a role in the processing of neuropeptide precursor proteins into their final bioactive forms. PMID- 11858718 TI - Enhancement of poplar glutaredoxin expression by optimization of the cDNA sequence. AB - Glutaredoxins are low-molecular-weight oxidore ductases that play an important role in redox regulation in eukaryotic and prokaryotic cells. Because of their low abundance, these proteins are poorly characterized in plants. Furthermore, very poor yields have been obtained with the expression systems prepared so far, and in addition, the recombinant products contain a His-tag which can interfere with the biochemical characterization. In order to obtain more information about those important regulatory proteins in plants, a cDNA coding for an extended glutaredoxin has been introduced into the expression plasmid pET-3d and the resulting construction has been used to transform Escherichia coli strain BL21(DE3) in the presence of plasmid helper pSBET or not. Initially poor or ineffective protein expression has been improved by successively cloning a N terminus truncated form of the protein, introducing silent mutations both at the 5' and at the 3' ends of the nucleotide sequence, and finally altering the 3' end in order to change the C-terminus amino acid sequence of the protein. The first modifications have allowed us to produce the protein in large amounts but essentially in an insoluble form which could be resolubilized and purified. On the other hand, changing the C-terminus sequence resulted in protein preparations of high purity and in a soluble form. The recombinant proteins were biochemically active and the yield varied between 6 and 14 mg of homogeneous protein per liter of culture. PMID- 11858719 TI - Identification of in vitro folding conditions for procathepsin S and cathepsin S using fractional factorial screens. AB - Human procathepsin S and cathepsin S were expressed as inclusion bodies in Escherichia coli. Following solubilization of the inclusion body proteins, fractional factorial protein folding screens were used to identify folding conditions for procathepsin S and cathepsin S. A primary folding screen, including eight factors each at two levels, identified pH and arginine as the main factors affecting procathepsin S folding. In a second simple screen, the yields were further improved. The in vitro folding of mature cathepsin S has never been reported previously. In this study we used a series of fractional factorial screens to identify conditions that enabled the active enzyme to be generated without the prodomain although the yields were much lower than achieved with procathepsin S. Our data show the power of fractional factorial screens to rapidly identify folding conditions even for a protein that does not easily fold into its active conformation. PMID- 11858720 TI - Requirement of continuous transcription for the synthesis of sufficient amounts of protein by a cell-free rapid translation system. AB - To understand the key processes of cell-free protein synthesis, the synthesis of adipose-type fatty acid binding protein (A-FABP) by a rapid translation system was examined under various conditions. The synthesis of A-FABP was achieved by using an expression vector of A-FABP containing a T7 promoter. However, synthesis of A-FABP was not observed when an RNA fragment corresponding to the open reading frame of A-FABP was used in the reaction instead of the expression vector. Northern analysis revealed that the RNA that was added to the reaction mixture promptly underwent degradation. On the contrary, when the expression vector of A FABP was employed, a strong RNA signal was observed over the entire incubation period. Thus, a continuous supply of RNA is needed in order to account for its loss via degradation to achieve the synthesis of reasonable amounts of A-FABP. Furthermore, the effect of continuous exchange of reaction mixture was also evaluated by measurement of the amount of synthesized A-FABP. PMID- 11858721 TI - Cloning and expression of human phenylalanyl-tRNA synthetase in Escherichia coli: comparative study of purified recombinant enzymes. AB - Human phenylalanyl-tRNA synthetase (PheRS) was cloned and expressed in Escherichia coli. The cDNAs of the alpha and beta subunits were cloned into pET 21b(+) and pET-28b(+) vectors. The 6x histidine-tagged (HT) plasmids pET 21_HTbeta, pET-28_HTalpha, and pET-28_HTbeta were constructed. Three different types of (alphabeta)(2) heterodimers of human PheRS carrying HT at the N-terminus of either of two alpha or beta subunits or simultaneously on both of them were overproduced and purified. The heterodimeric protein with HT appended to the N terminus of the beta subunit revealed no activity in the aminoacylation reaction as opposed to those with HT on the alpha subunit. It is known from the structure of the Thermus thermophilus Phe system that the N-terminal coiled-coil domain of the alpha subunit is involved in the binding of cognate tRNA(Phe). Our data demonstrate that a histidine-tagged N-terminal extension appended to the alpha subunit does not affect the kinetic parameters of tRNA(Phe) aminoacylation. Elimination of the HT from the alpha subunit by thrombin cleavage leads to nonspecific splitting of the enzyme that occurs in parallel to the main reaction. In addition to the tagged proteins the properly assembled heterodimer containing intact alpha and beta subunits free of HT was overproduced and purified. Aminoacylation activity of the overproduced human PheRS in the crude bacterial extract is two orders of magnitude higher than the corresponding activity in human placenta and the yield of the recombinant enzyme overproduced in E. coli is five times higher. PMID- 11858722 TI - Two-step purification of mitochondrial Hsp70, Ssc1p, using Mge1(His)(6) immobilized on Ni-agarose. AB - The most abundant mitochondrial homolog of Hsp70, Ssc1p, is involved in the import and folding of mitochondrial proteins. We have developed an easy and efficient method for purifying Ssc1p. Following a first step of anion exchange at pH 6.6, a column of Mge1(His)(6) immobilized on Ni(2+)-agarose provides an efficient second dimension that results in highly purified protein. The strong and specific interaction between Ssc1p and its cofactor protein, Mge1, ensures that primarily functional protein is isolated. Ssc1p purified by this method hydrolyzed ATP with a turnover rate of 0.3/min. The ATP hydrolysis was enhanced slightly by Mge1, about 5 times by Mdj1, and 12 times by both cofactors together. The CD spectrum of Ssc1p had a pattern and temperature dependence similar to those shown for other hsp70 homologs, with a midpoint of the major transition at approximately 70 degrees C. PMID- 11858723 TI - Expression, purification, and kinetic characterization of full-length human fibroblast activation protein. AB - Human fibroblast activation protein (FAP), an integral membrane serine protease, was produced in insect cells as a hexa-His-tagged protein using a recombinant baculovirus expression system. Two isoforms of FAP, glycosylated and nonglycosylated, were identified by Western blotting using an anti-His-tag antibody and separated by lectin chromatography. The glycosylated FAP was purified to near homogeneity using immobilized metal affinity chromatography and was shown to have both postprolyl dipeptidyl peptidase and postgelatinase activities. In contrast, the nonglycosylated isoform demonstrated no detectable gelatinase activity by either zymography or a fluorescence-based gelatinase activity assay. The kinetic parameters of the dipeptidyl peptidase activity for glycosylated FAP were determined using dipeptide Ala-Pro-7-amino-trifluoromethyl coumarin as the substrate. The k(cat) is 2.0 s(-1) and k(cat)/K(m) is 1.0 x 10(4) M(-1) s(-1) at pH 8.5. The pH dependence of k(cat) reveals two ionization groups with pK(a1) of 7.0 and pK(a2) of 11.0. The pH profile of k(cat)/K(m) yields similar results with pK(a1) 6.2 and pK(a2) 11.0. The neutral pK(a1) is associated with His at the active site. The basic pK(a2) might be contributed from an ionization group that is not involved directly in catalysis, instead associated with the stability of the active site structure. PMID- 11858724 TI - Affinity purification and characterization of the Escherichia coli molecular chaperones. AB - The molecular chaperones are a group of proteins that are effective in vitro and in vivo folding aids and show a well-documented affinity for proteins lacking tertiary structure. The molecular chaperones were induced from lon(-) Escherichia coli mutants, affinity purified with an immobilized beta-casein column, and assayed for refolding activity with thermally and chemically denatured carbonic anhydrase B (CAB). Chaperones were induced with three treatments: heat shock at 39 degrees C, heat shock 42 degrees C, and alcohol shock with 3% ethanol (v/v). Lysates were applied to an immobilized beta-casein (30 mg/g beads) column. After removing nonspecifically bound proteins with 1 M NaCl, the molecular chaperones were eluted with cold water or 1 mM Mg-ATP. The cold water and Mg-ATP eluates were analyzed by SDS-PAGE. Western analysis identified five E. coli molecular chaperones including DnaK, DnaJ, GrpE, GroEL, and GroES. The purity of eluted chaperones was 58% with cold water and 100% with Mg-ATP. Refolding denatured CAB in the presence of Mg-ATP resulted in a 97% recovery of heat-denatured CAB and a 68% recovery of chemically denatured CAB. The use of affinity matrices for the chaperone purification which are effective as in vitro folding aids will be presented. PMID- 11858725 TI - Expression of ayu (Plecoglossus altivelis) Pit-1 in Escherichia coli: its purification and immunohistochemical detection using monoclonal antibody. AB - The pituitary-specific transcription factor Pit-1 belongs to the family of POU domain proteins and is known to play an important role in the differentiation of pituitary cells. Here we report the complete nucleotide sequence of cDNA encoding Pit-1 from the brackish water fish, ayu (Plecoglossus altivelis). Nucleotide sequence analysis of 1910 bp of ayu Pit-1 cDNA revealed an open reading frame of 1074 bp that encodes a protein of 358 amino acids containing a POU-specific domain, POU homeodomain, and an STA (Ser/Thr-rich activation) transactivation domain. We inserted the coding region of Pit-1 cDNA, obtained by PCR, into a pET 20b(+) plasmid to produce recombinant Pit-1 in Escherichia coli BL21 (DE3) pLysS cells. Upon induction with isopropyl beta-D-thiogalactopyranoside, Pit-1 was expressed and accumulated as inclusion bodies in E. coli. The protein was then purified in one step by affinity chromatography on a nickel-nitrilotriacetic acid agarose column under denaturing conditions. This method yielded 0.7 mg of highly pure and stable protein per 200 ml of bacterial culture. A band of 40 kDa, resolved as recombinant ayu Pit-1 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, agrees well with the molecular mass calculated from the translated cDNA sequence. The purified recombinant Pit-1 was confirmed in vitro through Western blot analysis, using its monoclonal antibody. This monoclonal antibody detected Pit-1 in the nuclei of ayu developing pituitary by immunohistochemical reaction. It serves as a good reagent for the detection of ayu Pit-1 in situ. PMID- 11858726 TI - Production and purification of a recombinant Staphylococcal enterotoxin B vaccine candidate expressed in Escherichia coli. AB - An attenuated, recombinant form of Staphylococcus enterotoxin B (rSEB) was overexpressed in Escherichia coli under transcriptional control of the T7 promoter. The 28-kDa rSEB was partially purified from soluble, intracellular protein by tangential flow filtration and differential ammonium sulfate precipitation. The intermediate product was then further purified using low pressure liquid chromatography including hydrophobic interaction, cation exchange, and size-exclusion matrices. The final vialed product was >95% pure as determined by Coomassie blue-stained sodium dodecyl sulfate-polyacrylamide gel electrophoresis, high-pressure size-exclusion chromatography, and capillary zonal electrophoresis. The endotoxin level was <0.6 EU/mg. Final estimated yield of purified rSEB was 147 mg/L of starting culture. Purified rSEB was stable, elicited an immune response in mice, and protected mice against a lethal challenge with the native toxin. PMID- 11858727 TI - Self-immobilizing recombinant antibody fragments for immunoaffinity chromatography: generic, parallel, and scalable protein purification. AB - We present the directed immobilization of recombinant antibody fragments as ligands for general immunoaffinity chromatography methods. It is based on fusion proteins of scFv fragments with several chitin-binding domains which can be immobilized directly from a crude bacterial lysate on inexpensive chitin beads for the purification of proteins without any gradient or detector. It has been used with a positive pressure manifold, allowing the parallel processing of 24 different samples on a milligram scale, as convenient as plasmid isolation. The method is demonstrated with several anti-protein antibodies. In addition, methods are presented of using an anti-His tag antibody either alone or directly coupled to IMAC to obtain very pure protein. As those methods are scalable, they should prove very useful in the parallel purification of natural and recombinant proteins on small scales (for proteomics), medium scales (for crystallography and NMR), and very large scales (for therapeutic proteins). PMID- 11858728 TI - In vitro effects of polychlorinated biphenyls and hydroxy metabolites on nitric oxide synthases in rat brain. AB - Nitric oxide synthases (NOS) play a key role in motor activity in the cerebellum, hormonal regulation in the hypothalamus, and long-term potentiation (LTP), learning, and memory processes in the hippocampus. Developmental exposure to polychlorinated biphenyls (PCBs) has been shown to affect psychomotor functions, learning and memory processes, and to inhibit LTP. We hypothesized that PCBs may disrupt the regulation of such neurological functions by altering NOSs. We have studied the in vitro effects of several PCB congeners and some hydroxy PCBs on NOS activity in cytosolic (presumably neuronal NOS [nNOS]) and membrane (presumably endothelial NOS [eNOS]) fractions in different brain regions of young and adult rats. Among the two selected dichloro PCBs, the ortho-PCB, 2,2' dichlorobiphenyl (DCB), inhibited both cytosolic and membrane NOS activity at low micromolar concentrations (3-10 microM) in the selected brain regions of all age groups while the non-ortho-PCB, 4,4'-DCB, did not. 2,2'-DCB inhibited cytosolic NOS to a greater extent than membrane NOS. Pentachloro-PCBs (PeCBs) and hexachloro-PCBs (HCBs) did not have a significant effect on adult cerebellar cytosolic or membrane NOS. However, mono- and dihydroxy derivatives of HCBs significantly decreased cytosolic NOS (IC50s: 16.33 +/- 0.47 and 33.65 +/- 4.33 microM, respectively) but resulted in a marginal effect on membrane NOS in the cerebellum. Among three adult rat brain regions, the hypothalamic cytosolic NOS was the most sensitive to 2,2'-DCB. Also, cytosolic NOS in the cerebellum and hypothalamus of young rats was less sensitive than in the older rats. In summary, these results indicate that only di-ortho-PCB inhibited both NOS and hydroxy substitution of one or more chlorine molecules significantly increased the potency of both ortho- and non-ortho-HCBs. The selective sensitivity of NOS to dichloro- ortho-PCB and hydroxy metabolites suggests that the inhibition of NOS could play a role in the neuroendocrine effects as well as learning and memory deficits caused by exposure to PCBs. PMID- 11858729 TI - Increase of CYP1B1 transcription in human keratinocytes and HaCaT cells after UV B exposure. AB - Nonmelanoma skin cancers represent the most common malignant neoplasms in humans. UV-B play a major role in the etiology of these tumors, but exposure to environmental procarcinogens is also involved. CYP catalyzes numerous chemical carcinogen bioactivations and effects of UV-B on their expression are poorly understood. The aim of this study was to explore the molecular events involved in the induction of CYP1B1, a major cutaneous CYP, by UV-B. Our results demonstrated that unique UV-B exposure (20 mJ/cm(2)) increases human CYP1B1 transcript in primary keratinocytes and HaCaT cell cultures. Among 20 human samples studied, we observed a large interindividual variability of CYP1B1 mRNA induction (1.1- to 4.5-fold). Pretreatment with an antioxidant, N-acetylcysteine, repressed CYP1B1 increase, suggesting the involvement of UV-B photoproducts. alpha-Amanitin inhibition studies and CAT assays demonstrated that CYP1B1 mRNA induction is associated with a transcriptional activation of its expression. alpha Naphthoflavone inhibition studies and CAT assays performed after directed mutagenesis of xenobiotic responsive element sites showed the involvement of Ah receptor. Taken together, these data demonstrated that UV-B induces CYP1B1 gene expression after an activation of its transcription, which involves Ah receptor. PMID- 11858730 TI - The surface charge of visible particulate matter predicts biological activation in human bronchial epithelial cells. AB - The physicochemical complexity of airborne particulate matter (PM) has hampered identifying a specific mechanism(s) for its toxicity. In this study, selected physicochemical characteristics (i.e., size, particle number, acidity, and surface charge) were measured on various field PM, derived from urban ambient (St. Louis, Ottawa, Canada), residential (Woodstove), volcanic dust from Mt. St. Helen (MSH), and industrial [oil fly ash (OFA) coal fly ash (CFA)] sources. Morphometric analysis of visible (< or = 2.0 to >10 microm) field particles indicated that the industrial PM (OFA, CFA) had the smallest diameter and lowest total number of particles per weight while Woodstove and Ottawa had the largest diameter and highest number of particles. All PM lowered the pH of an unbuffered 10 mM NaCl solution from pH 7.4 to pH 4.7-6.8 but did not change the neutral pH of the cell culture medium, keratinocyte growth media (KGM). The surface charge (i.e., zeta potential) of microscopically visible (> or = 2.0 microm) field particles, suspended in either a Hepes-buffered KCl solution or in KGM, was measured by microelectrophoresis. In KCl solution, the mean zeta potential of all tested PM ranged from -36 +/- 2 (Woodstove) to -27 +/- 4.3 mV (MSH). When measured in KGM medium, the mean zeta potential value of each PM was significantly less (p > 0.001) than those measured in KCl solution, with values ranging from -17 +/- 0.3 mV (St. Louis) to -9 +/- 0.6 mV (MSH). Suspensions of field PM, its soluble and washed particulate fractions, were next prepared from each PM. The biological effects (i.e., increases in intracellular calcium ([Ca2+]i), cytokine release) of their exposure were measured in human, immortalized, tracheal-bronchial epithelial cells (BEAS-2B). Exposure of BEAS-2B cells to each fraction produced an immediate, but differential increase in [Ca2+]i and the subsequent release of the inflammatory cytokine IL-6, 4 and 16 h later. Increases in [Ca2+]i by field PM significantly correlated with the IL-6 released by each fraction (r2 > or = 0.76) after both 4 and 16 h exposures. The biological effects of each PM were compared with their physicochemical characteristics. No correlation was found between increases in [Ca2+]i or cytokine release and a PM's acidity or the number or size of its visible (> or = 2.0 microm) particles. However, the surface charge of PM field particles, when measured in the KGM exposure medium, showed a high correlation (r2 > or = 0.94) with the IL-6 release by field PM after both 4 and 16 h exposure. Increases in [Ca2+]i also correlated (r2 = 0.85) with the surface charge of PM field particles when measured in KGM. These data indicate that the surface charge (i.e., zeta potential) carried on PM's visible field particles predicts their differential release of the inflammatory cytokine IL-6 in cultures of human respiratory epithelial cells. PMID- 11858731 TI - Methamphetamine induces apoptosis in seminiferous tubules in male mice testis. AB - We investigated whether methamphetamine (MAMP) induces apoptosis in seminiferous tubules in 10-week-old male ICR mice. Methamphetamine was dissolved in saline and injected ip at four doses (1, 5, 10, and 15 mg/kg). TUNEL-positive cells were detected in the seminiferous tubules in animals 24 h after a single treatment with 5, 10, or 15 mg/kg MAMP. The percentage of seminiferous tubules containing more than three TUNEL-positive cells (apoptotic tubules) was considered a reliable indicator for apoptotic changes. After a single treatment with 10 mg/kg MAMP, the percentage of apoptotic tubules increased with time, and it became significant at 24 h, at which time vacuolar changes in spermatogonia also peaked. Although the percentage of apoptotic tubules increased dose dependently from 5 to 15 mg/kg MAMP, the DNA ladder was detected in the testis of 15 mg/kg MAMP-treated mice. Thus, we have demonstrated that MAMP induces apoptosis in seminiferous tubules in male mice testis. PMID- 11858732 TI - Increased glycogen content and glucose transporter 3 mRNA level in the placenta of Holtzman rats after exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin. AB - Exposure to a low dose of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) produces a variety of toxic manifestations, including fetal death. In order to evaluate the effects of low-dose TCDD on placental function in this study, pregnant Holtzman rats were given a single oral dose of 800 or 1600 ng TCDD/kg body wt or an equivalent volume of vehicle (control) on gestation day (GD) 15 and the results were observed on GD16 and GD20. The number of fetal deaths increased in the animals exposed to TCDD. Although fetal and placental weight did not differ significantly between the control group and the TCDD groups, histological differences from the control rats were clearly observed in the junctional zone (JZ) of the placentas of the TCDD-exposed rats. In the control placenta, glycogen cells occupied the majority of the JZ on GD16, but then decreased in number and almost disappeared by GD20, whereas on GD20 the placenta of the TCDD-exposed rats exhibited a larger area occupied by the glycogen cells and cysts filled with eosinophilic material surrounded by glycogen cells in the JZ than that of the control group. Glycogen assay revealed that the glycogen content of the placentas from the TCDD-exposed rats was higher than in the control rats. Semiquantitative RT-PCR analysis was performed to assess the expression of glucose transporter 1 (GLUT1) and GLUT3, the two major placental glucose transporter isoforms. On GD20 the level of expression of GLUT1 mRNA in the placentas was not different between the control and TCDD groups, whereas the level of expression of GLUT3 mRNA approximately doubled in both the 800 and 1600 ng/kg TCDD groups. GLUT3 mRNA expression was restricted to the labyrinth zone of placenta, where zone-specific expression of mRNA arylhydrocarbon receptor and induction of cytochrome P450 1A1 mRNA by TCDD were observed, and none was detected in the JZ. These results, including the increase of glycogen content and GLUT3 mRNA level in TCDD-exposed placentas, provide the first evidence of alteration of glucose kinetics in the placenta by TCDD. PMID- 11858733 TI - Modified endotoxin responses in rats pretreated with 1-->3-beta-glucan (zymosan A). AB - The present study investigates whether 1-->3-beta-glucans (zymosan particles) modify the pulmonary response of rats to endotoxin (lipopolysaccharide, LPS). Initial experiments were conducted to establish appropriate doses of LPS and regimens for exposure to zymosan and LPS. Interaction between zymosan and LPS exposures was determined to be the deviation from the sum of the individual effects of these agents. Treatment with zymosan on Day 1 and LPS on Day 2 modified several indices of pulmonary responsiveness, including tumor necrosis factor-alpha, albumin, and lactate dehydrogenase activity (LDH) in first acellular lavage fluid as well as the levels of chemiluminescence (CL), NO dependent CL, and nitric oxide production in cultured lavaged alveolar macrophage cells determined 1 day after exposure. No significant deviation from additivity was found for breathing rate increase and polymorphonuclear leukocytes infiltration. Simultaneous administration of zymosan and LPS or administration of LPS before zymosan did not change these indices of pulmonary responsiveness. These data suggest that the inhibitory effect of 1-->3-beta-glucans on pulmonary responsiveness to endotoxin exposure was apparent only when rats were pretreated with 1-->3-beta-glucan. These results suggest that complex interaction of components may exist in exposure to organic dusts. Therefore, hazard may not be defined by measuring endotoxin or 1-->3-beta-glucans alone. PMID- 11858734 TI - Noncholinergic actions of atropine on GABAergic synaptic transmission in the subfornical organ of rat slice preparations. AB - Actions of atropine on GABAergic inhibitory postsynaptic currents to neurons in the subfornical organ, which is a circumventricular organ, were studied by using rat slice preparations with whole-cell clamp recordings. Atropine at 0.01-1 microM antagonized the decreased frequency of Inhibitory postsynaptic currents by carbachol (Xu et al., Am. J. Physiol. Integr. Comp. Physiol. 280, R1657-R1664, 2001). It acted as a muscarinic antagonist at relatively low concentrations. Although the low concentrations of atropine did not change frequency and amplitude of the inhibitory postsynaptic currents, atropine at 10 microM to 1 mM did decrease them in a dose-dependent manner. Glutamatergic excitatory postsynaptic currents were not influenced by atropine at 100 microM. Atropine at 100 microM suppressed GABA- and muscimol-induced outward currents, but not kainic acid-induced inward currents. In addition, decrease of membrane conductance and induction of inward currents by 100 microM of atropine at a holding membrane potential, -51 mV, were found in subfornical organ neurons. From voltage-current curves, a mean reversal potential was estimated to be -65.9 +/- 3.7 mV, near to an equilibrium potential of chloride channels. These imply that atropine at 100 microM suppresses openings of chloride channels. Taken together, it is suggested that, while atropine at low concentrations has an antagonistic action on muscarinic responses, atropine at high concentrations suppresses GABAergic synaptic transmission in subfornical organ neurons. These findings may be of considerable value in understanding the central mechanisms of extraordinary drinking behavior in atropine intoxication. PMID- 11858735 TI - Liarozole markedly increases all trans-retinoic acid toxicity in mouse limb bud cell cultures: a model to explain the potency of the aromatic retinoid (E)-4-[2 (5,6,7,8-tetrahydro-5,5,8,8-tetramethyl-2-naphthylenyl)-1-propenyl] benzoic acid. AB - The remarkable toxicity of (E)-4-[2-(5,6,7,8-tetrahydro-5,5,8,8-tetramethyl-2 naphthylenyl)-1-propenyl] benzoic acid (TTNPB) compared to all trans-retinoic acid (tRA) is due to multiple factors, including reduced affinities for cytosolic binding proteins (CRABPs), resistance to metabolism, and prolonged nuclear receptor activation. To further investigate the role of half-life in retinoid toxicity, experiments were performed to determine whether, and to what extent, inhibition of tRA metabolism by liarozole increased its toxicity comparable to that of TTNPB in the mouse limb bud system. Liarozole is a known inhibitor of tRA 4-hydroxylation (CYP26). In the absence of liarozole, the IC50 for inhibition of chondrogenesis by tRA was 140 nM compared to 0.3 nM for TTNPB, a 467-fold difference. Following the addition of liarozole (10(-6) M) to limb bud cultures, the potency of tRA to inhibit chondrogenesis was increased approximately 14-fold (IC50 of 9.8 nM). Although liarozole markedly increased toxicity of tRA in mouse limb bud micromass cultures, tRA metabolism was inhibited only about 10%. These results indicate that a relatively minor decrease in the metabolism of tRA in the mouse limb bud system is associated with a marked enhancement of toxicity that is likely related to the prolongation of tRA half-life during a critical period of development. Thus, the prolonged half-life of TTNPB is the most significant factor contributing to the remarkable teratogenicity of this synthetic aromatic retinoid. PMID- 11858737 TI - Novel bioactive clerodane diterpenoids from the leaves and twigs of Casearia sylvestris. AB - Fractionation of a methanol extract of the leaves and twigs of Casearia sylvestris, as directed by activity against KB cell cytotoxicity, led to the isolation of three novel clerodane diterpenoids, casearvestrins A-C (1-3). The structures of 1-3 were deduced from one- and two-dimensional NMR experiments, including relative stereochemical assignments based on ROESY correlations and COSY coupling constants. All three compounds displayed promising bioactivity, both in cytotoxicity assays against a panel of tumor cell lines and in antifungal assays via the growth inhibition of Aspergillus niger in a disk diffusion assay. PMID- 11858738 TI - Structure and stereochemistry of new cytotoxic clerodane diterpenoids from the bark of Casearia lucida from the Madagascar rainforest. AB - Bioassay-guided fractionation of a CH(2)Cl(2)/MeOH extract of the bark of Casearia lucida resulted in the isolation of 11 new clerodane diterpenes, namely, casearlucins A-K (1-11), and three known clerodane diterpenoids, rel (2S,5R,6R,8S,9S,10R,18S,19R)-diacetoxy-18,19-epoxy-6-hydroxy-2-(2xi methylbutanoyloxy)cleroda-3,13(16),14-triene (12), rel (2S,5R,6R,8S,9S,10R,18S,19R)-18,19-diacetoxy-18,19-epoxy-6-methoxy-2-(2xi methylbutanoyloxy)cleroda-3,13(16),14-triene (13), and rel (2S,5R,8S,9S,10R,18S,19R)-18,19-diacetoxy-18,19-epoxy-2-(2xi methylbutanoyloxy)cleroda-3,13(16),14-triene (14). The structures of compounds 1 11 were established on the basis of extensive 1D and 2D NMR spectroscopic data interpretation. All compounds exhibited cytotoxicity activity against the A2780 ovarian cancer cell line, but none of the six compounds selected for testing in multiple cell lines showed significant selectivity. PMID- 11858739 TI - Emmyguyacins A and B: unusual glycolipids from a sterile fungus species that inhibit the low-pH conformational change of hemagglutinin A during replication of influenza virus. AB - Two novel glycolipids, emmyguyacin A (1a) and emmyguyacin B (1b), were isolated at concentrations of 1.51 g/L from a potato dextrose agar fermentation of a sterile fungus species. The compounds inhibit replication of influenza A virus (A/X31) in MDCK cells by inhibiting the pH-dependent conformational change of hemagglutinin A (IC(50) 9 microM). The structures were deduced using one- and two dimensional NMR techniques and mass spectrometric analyses on both the parent compounds and a host of degradation products and derivatives. A novel and unusual oxalic acid ester of a monohydroxylated fatty acid (5, 17-oxalyloxydocosanoic acid) is reported. The first isolation and characterization of the fatty acid 17 hydroxydocosanoic acid (3) itself is also reported as a saponification product of 1. PMID- 11858740 TI - Two major saponins from seeds of Barringtonia asiatica: putative antifeedants toward Epilachna sp. larvae. AB - Two major saponins have been isolated from a methanol extract of the seeds of Barringtonia asiatica, and their structures elucidated (mainly by two-dimensional NMR spectroscopy) as 3-O-[[beta-D-galactopyranosyl(1-->3)-beta-D-glucopyranosyl(1 ->2)]-beta-D-glucuronopyranosyloxy]-22-O-(2-methylbutyroyloxy)-15,16,28 trihydroxy-(3beta,15alpha,16alpha,22alpha)-olean-12-ene (3) and 3-O-[[beta-D galactopyranosyl(1-->3)-beta-D-glucopyranosyl(1-->2)]-beta-D glucuronopyranosyloxy]-22-O-[2(E)-methyl-2-butenyloyloxy]-15,16,28-trihydroxy (3beta,15alpha,16alpha,22alpha)-olean-12-ene (4). The antifeedant properties of 3 and 4 toward Epilachna larvae are discussed. PMID- 11858741 TI - Dictyterpenoids A and B, two novel diterpenoids with feeding-deterrent activity from the brown alga Dilophus okamurae. AB - In our continuing efforts to study the presence of biologically active compounds that control seaweed-herbivore interactions, we examined the presence of such chemicals in freshly collected Dilophus okamurae Dawson. Our results demonstrated the presence of two diterpenoids, named dictyterpenoid A (1) and dictyterpenoid B (2), having a novel carbon skeleton, along with 10 known diterpenes, 3-12. The structures of these novel metabolites were elucidated by spectroscopic and chemical methods. The isolated compounds showed feeding-deterrent activity against the young abalone Haliotis discus hannai. PMID- 11858742 TI - Structural and stereochemical reassessment of sclerophytin-type diterpenes. AB - A complete reassessment of the structural assignments of the sclerophytin diterpenes is provided. PMID- 11858743 TI - New cyanogenic and alkyl glycoside constituents from Phyllagathis rotundifolia. AB - Methanolic extracts of the leaves, stems, and roots of Phyllagathis rotundifolia collected in Malaysia yielded seven galloylated cyanogenic glucosides based on prunasin, with six of these being new compounds, prunasin 2',6'-di-O-gallate (3), prunasin 3',6'-di-O-gallate (4), prunasin 4',6'-di-O-gallate (5), prunasin 2',3',6'-tri-O-gallate (6), prunasin 3',4',6'-tri-O-gallate (7), and prunasin 2',3',4',6'-tetra-O-gallate (8). Also obtained was a new alkyl glycoside, oct-1 en-3-yl alpha-arabinofuranosyl-(1-->6)-beta-glucopyranoside (9). For compounds 3 8, the galloyl groups were individually linked to the sugar moieties via ester bonds. All new structures were established on the basis of NMR and MS spectroscopic studies. In addition, prunasin (1), gallic acid and its methyl ester, beta-glucogallin, 3,6-di-O-galloyl-D-glucose, 1,2,3,6-tetra-O-galloyl-beta D-glucose, strictinin, 6-O-galloyl-2,3-O-(S)-hexahydroxydiphenoyl-D-glucose, praecoxin B, and pterocarinin C were isolated and identified. The isolation of 1 and its galloyl derivatives (3-8) from a Melastomataceous plant are described for the first time. PMID- 11858744 TI - Efficient formal total synthesis of physostigmine and physovenine: conformational analysis of key intermediates. AB - An efficient route for the formal total synthesis of physostigmine (1) and physovenine (2), alkaloids from 5-methoxyindole-3-acetonitrile, through a Grignard reagent 1,4-addition, is described. 2-Hydroxyindolenine 5, the key advanced intermediate for the synthetic targets, was converted either to esermethole (12) via a high-yielding (28%) seven-step sequence or to the C-ring oxygenated analogue 15 in a five-step sequence and 23% overall yield. (1)H NMR and molecular modeling analyses of esermethole (12) and the furoindolines 13 and 15 were used to deconvolute weighted time-average vicinal coupling constants to provide definite solution-state conformational preferences in CD(2)Cl(2) solvent. PMID- 11858745 TI - Neoclerodane diterpenoids from Teucrium montbretii subsp. libanoticum and their absolute configuration. AB - From the aerial parts of Teucrium montbretii subsp. libanoticum 10 neoclerodane diterpenoids were isolated. Three of them are new [3beta-hydroxyteubutilin A (1), 12-epi-montanin G (2), 20-epi-3,20-di-O-deacetylteupyreinidin (3)], whereas the other seven, namely, 6-ketoteuscordin (4), teuscordinon (5), 6beta hydroxyteuscordin (6), montanin D (7), 3,20-di-O-deacetylteupyreinidin (8), montanin G (9), and 3-O-deacetylteugracilin A (10), are previously known structures. The structures of 1-3 were determined by spectral and chemical methods. PMID- 11858746 TI - Beesiosides G, H, and J-N, seven new cycloartane triterpene glycosides from Beesia calthifolia. AB - Seven new cycloartane glycosides (1-7), beesiosides G, H, and J-N, together with beesioside I (8) and beesioside A, were isolated from the rhizomes of Beesia calthifolia, and their structures were established by spectroscopic and chemical methods. Beesiosides G, H, and J-N were assigned as 20xi(1),24xi(2)-epoxy-9,19 cyclolanostane-3beta,16beta,18,25-tetraol-3-O-beta-D-glucopyranoside (1), 20xi(1),24xi(2)-epoxy-9,19-cyclolanostane-3beta,16beta,18,25-tetraol-3-O-[beta-D glucopyranosyl-(1-->6)]-beta-D-glucopyranoside (2), (20S,24R)-15alpha,16beta diacetoxy-20,24-epoxy-9,19-cyclolanostane-3beta,18,25-triol-3-O-beta-D xylopyranoside (3), (20S,24S)-16beta-acetoxy-18,24;20,24-diepoxy-9,19 cyclanostane-3beta,15beta,25-triol-3-O-beta-D-xylopyranoside (4), (20S,24S) 16beta-acetoxy-18,24;20,24-diepoxy-9,19-cyclanostane-3beta,25-diol-3-O-beta-D xylopyranoside (5), 20xi(1),24xi(2)-epoxy-15alpha-acetoxy-9,19-cyclolanostane 3beta,16beta,25-triol-3-O-beta-D-xylopyranoside (6), and 20xi(1),24xi(2)-epoxy 9,19-cyclolanostane-3beta,12alpha,15alpha,16beta,25-pentaol-3-O-beta-D xylopyranoside (7), respectively. PMID- 11858747 TI - Antifungal steroidal glycosides from the patagonian starfish anasteriasminuta: structure-activity correlations. AB - Two new sulfated steroidal hexaglycosides, anasterosides A (2) and B (3), along with the known versicoside A (1) have been isolated from the Patagonian starfish Anasterias minuta. Their structures have been elucidated by spectroscopic analysis (NMR and FABMS) and chemical transformations. Compounds 1 and 2 and the synthetic pentaglycoside 1b derived from versicoside A showed antifungal activity against the plant pathogenic fungus Cladosporium cucumerinum. Desulfation of hexaglycoside 1 rendered a totally inactive saponin. PMID- 11858748 TI - Eupha-7,9(11),24-trien-3beta-ol ("antiquol C") and other triterpenes from Euphorbia antiquorum latex and their inhibitory effects on Epstein-Barr virus activation. AB - The structures of three triterpene alcohols isolated from the latex of Euphorbia antiquorum were established to be eupha-7,9(11),24-trien-3beta-ol (2; antiquol C), 19(10-->9)abeo-8alpha,9beta,10alpha-eupha-5,24-dien-3beta-ol (3; antiquol B), and 24-methyltirucalla-8,24(24(1))-dien-3beta-ol (4; euphorbol) on the basis of spectroscopic methods. Compounds 3 and 4 have previously been assigned the erroneous structures of 10alpha-cucurbita-5,24-dien-3alpha-ol and 24-methyleupha 8,24(24(1))-dien-3beta-ol, respectively. Compounds 2-4 and four other known compounds isolated from the latex, euphol (1), lemmaphylla-7,21-dien-3beta-ol (5), isohelianol (6), and camelliol C (7), showed potent inhibitory effects on Epstein-Barr virus early antigen (EBV-EA) activation induced by the tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA). PMID- 11858749 TI - Constituents of the bark and twigs of Artocarpus dadah with cyclooxygenase inhibitory activity. AB - Fractionation of an ethyl acetate-soluble extract of the bark of Artocarpus dadah has led to the isolation of three new prenylated stilbenoid derivatives, 3 (gamma,gamma-dimethylallyl)resveratrol (1), 5-(gamma,gamma dimethylallyl)oxyresveratrol (2), 3-(2,3-dihydroxy-3-methylbutyl)resveratrol (3), and a new benzofuran derivative, 3-(gamma,gamma-dimethylpropenyl)moracin M (4), along with six known compounds, oxyresveratrol, (+)-catechin, afzelechin-3-O alpha-L-rhamnopyranoside, (-)-epiafzelechin, dihydromorin, and epiafzelechin (4beta-->8)-epicatechin. From an ethyl acetate-soluble extract of the twigs of the same plant were isolated compound 4 and two new neolignan derivatives, dadahols A (5) and B (6), as well as 10 known compounds, oxyresveratrol, (+) catechin, afzelechin-3-O-alpha-L-rhamnopyranoside, resveratrol, steppogenin, moracin M, isogemichalcone B, gemichalcone B, norartocarpetin, and engeletin. The structures of compounds 1-6 were determined using spectroscopic and chemical methods. Isolates were evaluated for their inhibitory effects against both cyclooxygenase-1 (COX-1) and -2 (COX-2) and in a mouse mammary organ culture assay. PMID- 11858750 TI - Bioactive saponins from Acacia tenuifolia from the suriname rainforest. AB - Bioassay-guided fractionation of the MeOH extract of Acacia tenuifolia using the engineered yeast strains 1138, 1140, 1353, and Sc7 as the bioassay tool resulted in the isolation of the three new saponins 3, 5, and 6 and the three known saponins 1, 2, and 4. The structures of the new compounds were established on the basis of HRMS, 1D and 2D NMR spectral data on the intact saponins, and GC-MS analyses of the sugars. Compounds 1,2 and 5,6 showed cytotoxicity against mammalian cell lines. PMID- 11858751 TI - Flavonoids from Ulex airensis and Ulex europaeus ssp. europaeus. AB - From the dichloromethane extract of Ulex airensis three new isoflavonoids, ulexin C (1), ulexin D (2), and 7-O-methylisolupalbigenin (3), were isolated and characterized by spectroscopic methods. Ulexin D (2) was also identified from the dichloromethane extract of Ulex europaeus ssp. europaeus. Together with these new metabolites, 18 compounds of previously known structures were isolated and identified from both species. The antifungal activity of these compounds was tested against Cladosporium cucumerinum by a bioautographic TLC assay. PMID- 11858752 TI - Anti-inflammatory and anti-tumor-promoting effects of cucurbitane glycosides from the roots of Bryonia dioica. AB - Seven new triterpene glycosides, bryoniosides A-G (1-7), have been isolated along with two known triterpene glycosides, cabenoside D (8) and bryoamaride (9), from a methanol extract of the roots of Bryoniadioica. The structures of 1-7 were determined on the basis of spectroscopic and chemical methods. Six compounds, 2, 3, 5, and 7-9, and 11 compounds, 1-9, bryodulcosigenin (10), and bryosigenin (11), respectively, were evaluated for their inhibitory effects on 12-O tetradecanoylphorbol-13-acetate (TPA)-induced inflammation (1 microg/ear) in mice and on Epstein-Barr virus early antigen (EBV-EA) activation induced by TPA. All compounds tested showed marked anti-inflammatory effects, with 50% inhibitory doses (ID(50)) of 0.2-0.6 mg per ear. In addition, all of the compounds tested except for compound 5 showed potent inhibitory effects on EBV-EA induction (100% inhibition at 1 x 10(3) mol ratio/TPA). PMID- 11858753 TI - Deoxyartemisinin derivatives from photooxygenation of anhydrodeoxydihydroartemisinin and their cytotoxic evaluation. AB - Photooxygenation of anhydrodeoxydihydroartemisinin (4) followed by chromatographic separation of the reaction mixture yielded the new compounds alpha- (5) and beta-hydroperoxydeoxyartemisitene (8) and the formate ester 7, together with two previously reported compounds, 6 and 9. Reduction of 5 using polymer-bound triphenylphosphine afforded the new compound dihydrodeoxyartemisitene (10). Treatment of 10 with a catalytic amount of BF(3) OEt(2) yielded the C(2)-symmetrical dimer bis(dihydrodeoxyartemisitene) ether (11) and two new compounds, dihydrodeoxyartemisitene methyl ether (12) and the dimer 13, as minor products. Dehydroacetoxylation of 5 using acetic anhydride in pyridine afforded deoxyartemisitene (14). The identities of the new compounds (5, 7, 8, 10-14) were deduced from their spectral data and by chemical derivatization. The stereochemistry of dimer 11 was defined on the basis of X-ray crystallographic analysis. All compounds were evaluated in vitro in the National Cancer Institute drug-screening program consisting of 60 human cancer cell lines derived from nine different tissues. Of the compounds tested, deoxyartemisitene (14) demonstrated significant cytotoxicity against a number of human cancer cell lines. PMID- 11858754 TI - Cytotoxic effect (on tumor cells) and in vitro antiviral activity against herpes simplex virus of synthetic spongiane diterpenes. AB - A series of synthetic spongiane-type diterpenes have been tested in vitro for their potential antitumor and antiherpetic activity. Although the antiviral activity of these compounds against herpes simplex virus type 2 (HSV-2) was very weak, some compounds exhibited relevant cytotoxicity in the human tumor cell lines HeLa and HEp-2. The biological activity of formyl spongianes is reported for the first time. With the present study, some structure-activity trends are suggested for the cytotoxic activity of these sponge-derived natural products. PMID- 11858755 TI - Novel acylated triterpene glycosides from Muraltia heisteria. AB - Four new acylated triterpene glycosides (1-4) have been isolated as two inseparable mixtures of the trans- and cis-3,4,5-trimethoxycinnamoyl derivatives (1,2 and 3,4) from the roots of Muraltia heisteria. The structures of these compounds were elucidated by various 1D and 2D NMR techniques, including (1)H and (13)C, COSY, NOESY, HSQC, TOCSY, and HMBC experiments and FABMS. Compounds 3 and 4 were shown to be cytotoxic in a human colon cancer cell line but did not show any ability to potentiate in vitro cisplatin cytotoxicity. PMID- 11858756 TI - New sugar-mimic alkaloids from the pods of Angylocalyx pynaertii. AB - Chromatographic separation of the pod extract of Angylocalyx pynaertii resulted in the isolation of 13 sugar-mimic alkaloids (1-13). The structures of the new alkaloids were elucidated by spectroscopic methods as the 6-O-beta-D-glucoside (10) and N-hydroxyethyl derivative (11) of 1,4-dideoxy-1,4-imino-D-arabinitol (DAB) (1), 1,6-dideoxynojirimycin (12), and 1,3,4-trideoxynojirimycin (13). 2,5 Imino-1,2,5-trideoxy-L-glucitol (7), 2,5-dideoxy-2,5-imino-D-fucitol (8), and beta-homofuconojirimycin (9), isolated from the pods as well as the bark, were very specific inhibitors of alpha-L-fucosidase with no significant inhibitory activity toward other glycosidases. In this work, 1,4-dideoxy-1,4-imino-D-ribitol (6) was found to be a better inhibitor of lysosomal beta-mannosidase than 2,5 imino-1,2,5-trideoxy-D-mannitol (2). N-Hydroxyethyl-1-deoxynojirimycin (miglitol), which is commercially available for the treatment of diabetes, retained its inhibitory potential toward rat intestinal maltase and sucrase, whereas 11 and the synthetic N-hydroxyethyl derivative of 2,5-dideoxy-2,5-imino-D mannitol markedly lowered or abolished their inhibition toward all enzymes tested. PMID- 11858757 TI - Achyrofuran, a new antihyperglycemic dibenzofuran from the South American medicinal plant Achyrocline satureioides. AB - A new prenylated dibenzofuran, achyrofuran (2), was isolated from an extract of Achyrocline satureioides by bioassay-guided fractionation using the db/db mouse model for type 2 diabetes. Compound 2significantly lowered blood glucose levels in this model when administered orally at 20 mg/kg q.d. PMID- 11858758 TI - Lignanamides and nonalkaloidal components of Hyoscyamus niger seeds. AB - Four lignanamides, a tyramine derivative, and 10 other nonalkaloidal components were isolated from the seeds of Hyoscyamus niger. Among them, hyoscyamide (1), 1,24-tetracosanediol diferulate (6), and 1-O-(9Z,12Z-octadecadienoyl)-3-O nonadecanoyl glycerol (7) are new structures. The other compounds were identified as grossamide, cannabisin D, cannabisin G, N-trans-feruloyl tyramine, 1-O octadecanoyl glycerol, 1-O-(9Z,12Z-octadecadienoyl) glycerol, 1-O-(9Z,12Z octadecadienoyl)-2-O-(9Z,12Z-octadecadienoyl) glycerol, 1-O-(9Z,12Z octadecadienoyl)-3-O-(9Z-octadecenoyl) glycerol, rutin, vanillic acid, beta sitosterol, and daucosterol. Grossamide, and cannabisins D and G exhibited moderate cytotoxicity in cultured LNCaP human prostate cancer cells. PMID- 11858759 TI - Bioactive isomalabaricane triterpenes from the marine sponge Rhabdastrella globostellata. AB - Two new isomalabaricane triterpenes, stellettin H (1) and stellettin I (2), have been isolated from the marine sponge Rhabdastrella globostellata, collected from the Philippines. Stellettins A-D (3-6), (-)-stellettin E (7), and rhabdastrellic acid-A (8) were also isolated and characterized. Stellettin B (4) and (-) stellettin E (7) showed selective cytotoxicity toward p21(WAF1/Cip1)-deficient human colon tumor (HCT-116) cells with IC(50) values of 0.043 and 0.039 microM, respectively. PMID- 11858760 TI - Pseudoirroratin A, a new cytotoxic ent-kaurene diterpene from Isodon pseudo irrorata. AB - A new ent-kaurene diterpene, pseudoirroratin A (1), and a known diterpene, pseurata A, were isolated from Isodon pseudo-irrorata. Compound 1 showed significant cytotoxicity against the Lu1, SW626, LNCaP, KB, and HOS cancer cell lines with IC(50) values of 0.26 (0.75), 0.20 (0.57), 0.90 (2.59), 0.90 (2.59), and 0.50 (1.44) microg/mL (microM), respectively. The structure of 1 was elucidated by spectroscopic means including 1D and 2D NMR techniques. PMID- 11858761 TI - Three new homoisoflavanone glycosides from the bulbs of Ornithogalum caudatum. AB - Phytochemical examination of Ornithogalum caudatum led to the isolation of three new homoisoflavanone glycosides. Their structures were elucidated, on the basis of the spectroscopic data and chemical evidence and by comparing them with those of known compounds, as (-)-7-O-methyleucomol 5-O-beta-D-glucopyranoside (1), (-) 7-O-methyleucomol 5-O-beta-rutinoside (2), and (-)-7-O-methyleucomol 5-O-beta neohesperidoside (3), respectively. PMID- 11858762 TI - Choosing the best pulse sequences, acquisition parameters, postacquisition processing strategies, and probes for natural product structure elucidation by NMR spectroscopy. AB - The relative merits of different pairs of two-dimensional NMR pulse sequences (COSY-90 vs COSY-45, NOESY vs T-ROESY, HSQC vs HMQC, HMBC vs CIGAR, etc.) are compared and recommendations are made for the preferred choice of sequences for natural product structure elucidation. Similar comparisons are made between different selective 1D sequences and the corresponding 2D sequences. Many users of 2D NMR use longer than necessary relaxation delays and neglect to use forward linear prediction processing. It is shown that using shorter relaxation delays in combination with forward linear prediction allows one to get better resolved spectra in less time. The relative merits of different probes and likely future probe developments are also discussed. PMID- 11858764 TI - Delta-6 desaturase activity and gene expression, tissue fatty acid profile and glucose turnover rate in hereditary hypertriglyceridemic rats. AB - OBJECTIVE: We have shown previously that the impaired insulin action in hereditary hypertriglyceridemic (hHTg) rat is accompanied by a specific fatty acid (FA) profile in the insulin target tissues, possibly due to a desaturation defect. Thus, the aim of this study was to measure the enzymatic activity and gene expression of delta-6 desaturase in liver of hHTg rats and the tissue FA composition in relation to insulin action. METHODS: Glucose, triglycerides and insulin in plasma were measured using commercially available enzymatic sets. The hepatic delta-6 desaturase activity in hHTg rats was determined radiometrically in a microsomal fraction using the 1-14C-linoleic acid as substrate. delta-6 Desaturase gene expression was measured by the Northern blot technique using a specific cDNA probe. Tissue FA profile was determined by gas chromatography in the total lipid fraction extracted to chloroform. The glucose turnover rate was measured in conscious freely moving animals with the aid of euglycemic hyperinsulinic clamp method. RESULTS: Tissue triglycerides showed a high accumulation in skeletal muscle of hHTg rats. In the liver of these animals, a defect in delta-6 desaturase enzymatic activity was found, while the gene expression for delta-6 desaturase was not changed. Such decreased delta-6 desaturase activity in the liver was linked to a decrease of delta-6 desaturase index as calculated from the liver FA composition. Also the concentration of arachidonic acid (a final metabolite in the biosynthesis of polyunsaturated fatty acids of the n-6 series) was significantly decreased in hHTg rat liver. These changes in FA metabolism were accompanied by a decreased glucose infusion rate (a measure of in vivo insulin action) required to maintain euglycemia at hyperinsulinemia in hHTg rats, and correlated with the hepatic delta-6 desaturase activity. CONCLUSIONS: 1. hHTg rats showed a reduced activity of the delta-6 desaturase in liver without any changes in gene expression for this enzyme; 2. such impairment is accompanied by a lower delta-6 desaturase index (18:2n-6/18:3n 6) found in the liver of these animals and by specific FA profiles in the tissues, particularly regarding the amount of long-chain PUFAs and 18:2n-6 metabolites; and (4) these alterations seem to be related to the impaired insulin action of hHTg rats. PMID- 11858765 TI - Single dose of morphine influences plasma corticosterone and gene expression of main NMDA receptor subunit in the adrenal gland but not in the hippocampus. AB - OBJECTIVE: This study was aimed to evaluate the effects of morphine on hypothalamo-pituitary-adrenocortical (HPA) axis, namely proopiomelanocortin (POMC) mRNA and plasma corticosterone, in relation to its influence on glutamate receptor gene expression in central and peripheral sites related to HPA axis regulation. As previous data on morphine action were obtained mainly in male rats, these experiments were performed in females to see potential gender differences. METHODS: Adult female Sprague-Dawley rats were injected with a single dose of morphine (10 mg/kg s.c.) or vehicle. Blood and tissues were sampled 4 h and 24 h following the treatment. In situ hybridization was used to measure POMC mRNA concentrations, reverse transcription-polymerase chain reaction to quantify mRNA coding for N-methyl-D-aspartic acid (NMDA) receptor subunit 1 and radioimmunoassay to measure plasma corticosterone. RESULTS: Single dose of morphine was followed by a decrease in gene expression of glutamate receptor subunit NMDAR1 in the adrenal gland. Concentrations of mRNAs coding for NMDAR1 in the hippocampus and for POMC in the anterior pituitary remained unaffected. However, plasma corticosterone levels, which were measured at 4 and 24 h after the treatment with morphine, showed a disturbed daily variation in corticosterone release. The efficacy of morphine was confirmed by Straub tail response, one of the classical effects of this drug, in mice. CONCLUSIONS: Present data obtained in females allow to suggest that morphine exerts some of its effects on HPA axis by POMC unrelated mechanisms seemingly in a gender specific manner. Decrease in glutamate receptor gene expression in adrenals induced by a single dose of morphine may result in a modulation of adrenal function in response to subsequent exposure to opioids and contribute to some alterations occurring during opioid drug abuse. PMID- 11858766 TI - Gene expression of catecholamine synthesizing enzymes in A5 cell group and modulation of tyrosine hydroxylase mRNA by immobilization stress. AB - OBJECTIVE: The A5 group of noradrenergic neurons plays a key role in autonomic mechanisms like cardiovascular regulation, nociception and respiration. The aim of this work was to detect the gene expression of catecholamine synthesizing enzymes in A5 brain nuclei. METHODS: The gene expression of. tyrosine hydroxylase (TH), dopamine-beta-hydroxylase (DBH) and phenylethanolamine N-methyl-transferase (PNMT) in A5 brain nuclei was estimated. We also investigated various time intervals after the end of the single two-hour immobilization, as well as the effect of short-term repeated immobilization (120 min daily for 7 days) on tyrosine hydroxylase gene expression, the rate-limiting enzymes in catecholamines biosynthesis, in the A5 cell group. For all experiments, reverse transcription with subsequent polymerase chain reaction (RT-PCR) was used. RESULTS: As expected, we detected a clear signal for TH and DBH mRNA but no signal for PNMT mRNA. Both, single and repeated immobilization stress exposure increased significantly the gene expression of TH in A5 area. Maximal elevation in TH mRNA levels occurred after single immobilization for two hours and subsequent decapitation 24 hours later. CONCLUSIONS: In this study we detected for the first time the presence of DBH mRNA in micro dissected A5 cell group. We also showed how the gene expression of tyrosine hydroxylase changed with the function of time after the single immobilization exposure. Thus, TH mRNA in A5 cell group is modulated by immobilization stress in a time-dependent manner. PMID- 11858767 TI - Serum insulin/like growth factor I, bone mineral density and biochemical markers of bone metabolism in children with idiopathic osteoporosis. AB - OBJECTIVE: To determine whether the serum concentration of insulin-like growth factor I (IGF-I) correlates with the occurrence of idiopathic osteoporosis in children, and whether serum levels of IGF-I correlate with selected bone metabolism markers in patients with osteoporosis. METHODS: The study comprised 24 patients aged 7-18 years, including 12 with idiopathic osteoporosis and 12 control children. Bone mineralisation disorders were diagnosed on the basis of complex clinical, densitometric and biochemical evaluation. In all children serum concentration of IGF-I was estimated by radioimmunoassay and the third fraction of IGF binding proteins (IGFBP3) by immunoradiometry. In children with osteoporosis the indices of bone metabolism were also assessed, e.g. serum levels of osteocalcin and activity of bone isoenzyme of alkaline phosphatase (bone formation markers) and urine concentration of pyridinoline and deoxypyridinoline and collagen type I crosslinked C-telopeptyde (resorption markers). RESULTS: It was found that in children with osteoporosis IGF-I concentration was significantly lower than in the control group (mean values were 583 and 850 ng/ml, respectively; P<0.05). These differences were independent on biological age of the studied children and were present in all adolescence stages. Concentrations of IGFBP3 did not differ significantly between groups (3593 vs. 3955 ng/ml), while that of IFG-I correlated positively with total and spinal bone mineral density (R=0.85 and R=0.80, respectively; P<0.00001). In children with osteoporosis there was also a significant relationship between IGF-I concentration and elimination of pyridinoline and deoxypyridinoline with urine (R=0.64 and R=0.65, respectively; P<0.05). There was no significant correlation between the concentration of IGF-I and IGFBP3 and other studied bone metabolism markers. CONCLUSIONS: The conducted study revealed that lower IFG-I concentrations correlate with higher bone resorption markers values and decreased mineralisation. These results suggest the importance of insulin-like growth factor in the ethiopathogenesis of idiopathic osteoporosis, which needs to be confirmed in further studies. PMID- 11858769 TI - Circadian serum levels of dehydroepiandrosterone sulphate in postmenopausal asthmatic women before and after long-term hormone replacement. AB - OBJECTIVE: To assess mean 24-h serum concentrations of dehydroepianrosterone (DHEAS) in postmenopausal women with asthma before and after hormone replacement therapy (HRT). METHODS: Studies were performed in 55 asthmatic and 20 healthy postmenopausal women aged 48-60 before HRT and after 6 months of transdermal 17b estradiol (E2) and medroxyprogesterone acetate treatment (cyclical method). Serum DHEAS concentrations were assessed with the use of RIA method. RESULTS: In the group of postmenopausal asthmatic women treated with glucocorticoids the mean 24 h DHEAS serum levels were lower than in a similar group not treated with glucocorticoids and a control group of healthy postmenopausal women. However, in both groups of asthmatic women (e.g. glucocorticoid treated and untreated) a significant increase of mean daily DHEAS levels after 6 months of HRT was observed. The hormone concentrations did not change in control group. CONCLUSIONS: Postmenopausal asthmatic women show diminished circadian dehydroepiandrosterone sulphate serum concentrations irrespective whether they were treated with glucocorticoids or not. However, after 6 months of hormonal replacement therapy in these groups increased levels of DHEA were found. PMID- 11858768 TI - Degeneration and restoration of spermatogenesis in relation to the changes in Leydig cell population following ethane dimethanesulfonate treatment in adult rats. AB - OBJECTIVE: To investigate degeneration and restoration patterns of spermatogenesis in relation to the changes in Leydig cells (LCs) after treatment with ethane dimethanesulfonate (EDS). MATERIALS AND METHODS: Adult Wistar male rats were treated with EDS at a dose 75 mg/kg body weight and the testes were sampled at 7, 14, 21, 35 and 49 days after treatment for histological and ultrastructural studies. RESULTS: During the first two weeks after treatment stage dependent loss of germ cells was found within seminiferous tubules that led to a profound disturbance of spermatogenesis. The restoration of seminiferous epithelium followed also in stage specific manner and in relation to development of a new LC population (third week). The development of new LCs after EDS treatment repeats the normal dynamics of postnatal LC development within a similar time range. CONCLUSION: EDS treatment of rats causes a temporary germ cell degeneration in the testis. The kinetics of disappearance of germ cells and their regeneration broadly follows the changes in LC population. PMID- 11858770 TI - Estrous cycle dependent changes in steroid secretion by pig ovarian cells in vitro to polychlorinated biphenyl (PCB 153). AB - OBJECTIVE: To characterize PCB 153 action on the ovary, the direct effect of PCB153 was investigated in vitro using a co-culture of pig granulosa and theca cells collected during different stages of follicular development. METHODS: The cells were cultured in the absence or presence of 5, 10, 50 or 100 ng/ml of PCB 153. Media were changed after 48, 96 and 144 h and frozen until further estradiol (E2), progesterone (P4) and testosterone (T) analysis. RESULTS: 48 hrs exposure of follicular cells collected from small-size follicles to all the investigated doses of PCB 153 caused statistically significant decrease in progesterone (P4) secretion and at doses of 50 ng and 100 ng/ml in testosterone (T) secretion. No effect on estradiol (E2) secretion was observed. After 96h and 144h exposure to PCB an increase in P4 secretion with concomitant drastic decrease in T secretion and a tendency to decrease in E2 secretion was observed. Similarly as in the case of small follicles, the action of PCB on steroid secretion by cells collected from medium follicles depended on time of exposure. The increase in T secretion and no influence on P4 and E2 secretion was observed after 2 days of exposure to PCB. Antiestrogenic action of PCB was noted after 4 and 6 days of exposure to PCB. In large, preovulatory follicles 2 days exposure to PCB had no effect on steroids secretion while longer exposition to this congener caused statistically significant antiestrogenic action. CONCLUSION: The presented paper suggests various actions of PCB 153 as an endocrine disrupter on estradiol, progesterone and testosterone secretion from ovarian cells in vitro which were dependent on both exposure length and stage of the follicular development. PMID- 11858771 TI - Minireview: microdialysis of the blood outflowing from the brain. AB - In past several years the in vivo blood microdialysis technique has been widely used for a variety of pharmacological and physiological applications to study, monitor and analyze endogenous substances, such as neurohormones, and exogenous substances such as therapeutic drugs and their metabolites. The technique is being described in detail and discussed, in which microdialysis probes were implanted into the jugular vein, blood was flowing freely around the dialysis membrane. The probe was perfused at a very low flow rate (approximately 1-2 microL/min)with the solution resembling closely the composition of body fluid. In this laboratory (Department of Experimental and Clinical Physiology, Institute of Physiology and Biochemistry, Medical University of Lodz) the technique of in vivo blood minidialysis was worked out in small laboratory animals (rat, guinea-pig, hamster) and used to demonstrate that neurohypophysial hormones can be released into the blood outflowing from the region of the sella turcica and blood dialysate from the femoral vein. PMID- 11858772 TI - Prevention of Left Ventricular Remodeling After Myocardial Infarction. AB - Postinfarction left ventricular remodeling begins early after acute myocardial infarction and may continue for months to years afterward. Early re-establishment of flow in the occluded artery is associated with smaller left ventricular cavity volumes and reduced remodeling. Acute percutaneous coronary intervention (PCI) or thrombolytic therapy (for patients more than 1 hour away from a catheterization facility) as early as possible after symptoms is critical. Late reperfusion (PCI more than 12 hours after infarction) may prove useful, and this will be determined by the results of ongoing clinical trials. Recurrent MI is reduced by antiplatelet agents (aspirin in most patients) and by 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors. Intravenous nitroglycerin may limit early (initial 24 hours) dilatation following infarction, but long-term use in asymptomatic patients is not efficacious. Beta- adrenergic receptor antagonists and angiotensin-converting enzyme (ACE) inhibitors have independent efficacy in attenuating the early and late phases of remodeling. The combined use of a beta blocker and an ACE inhibitor has greater efficacy than either agent alone, provided they are tolerated hemodynamically. Although angiotensin II receptor antagonists have similar efficacy to ACE inhibitors and have fewer side effects, the angiotensin II receptor blockers should be reserved for patients intolerant to ACE inhibitors. In patients requiring diuretic therapy, spironolactone is preferred because of its salutary properties regarding extracellular matrix remodeling, specifically in reducing fibrosis. Surgical revascularization with or without associated mitral valve repair is useful in selected patients with severe ischemic mitral regurgitation or hibernating myocardium. New therapies directed at modulating the remodeling process may focus on manipulating the components of the extracellular matrix to reduce the deleterious impact of this process. PMID- 11858773 TI - Restenosis: Intracoronary Brachytherapy. AB - Though interventional strategies have revolutionized the management of patients with symptomatic coronary artery disease, in-stent restenosis has emerged as the single most important limitation of long-term success following percutaneous coronary intervention. Once present, in-stent restenosis is extraordinarily difficult to treat, with conventional revascularization techniques failing in 50% to 80% of patients. Intracoronary radiation, or brachytherapy, targets cellular proliferation within the culprit neointima. Clinical trials have demonstrated that brachytherapy is a highly effective treatment for in-stent restenosis, reducing angiographic restenosis by 50% to 60% and the need for target vessel revascularization by 40% to 50%. The benefits of intracoronary brachytherapy may be particularly pronounced in certain patient subgroups (eg, those with diabetes, long lesions, or lesions in saphenous vein bypass grafts), but comes at the cost of an increased rate of late stent thrombosis and the need for extended antiplatelet therapy. The role of brachytherapy in the arsenal of the interventional cardiologist will continue to evolve, particularly in light of the unprecedented recent advances with the use of drug-eluting stents for restenosis prevention. PMID- 11858774 TI - Familial Hypercholesterolemia. AB - Heterozygous familial hypercholesterolemia is a common, severe form of elevated plasma, low-density lipoprotein (LDL) cholesterol. Familial hypercholesterolemia is under-recognized and under-treated despite well-known benefits from available lipid-lowering medications. To help address these problems, the humanitarian MEDPED (Make Early Diagnoses, Prevent Early Deaths) organization was developed as a means to find and help persons with familial hypercholesterolemia all over the world. Although some patients with familial hypercholesterolemia can be treated adequately with a statin alone, many require aggressive treatment with a combination of drugs, including a statin, bile acid sequestrant, and niacin to reach LDL goals. Such treatment has resulted in regression of angiographically monitored coronary lesions in patients with familial hypercholesterolemia. PMID- 11858775 TI - Coronary Heart Disease in Women: Hormone Replacement Therapy. AB - For both primary and secondary prevention of coronary heart disease (CHD), it is prudent to use strategies that are of proven benefit and that do not harm patients. In all women, these strategies include lifestyle approaches such as smoking avoidance, proper nutrition, and regular exercise. Lipid-lowering and blood pressure control with pharmacotherapy are indicated in women who do not meet target lipid or blood pressure levels with lifestyle interventions. For women with CHD, aspirin, beta-blockers, and angiotensin-converting enzyme inhibitors should be considered. Widespread under-use of established preventive therapies has been documented in women. These interventions should be emphasized in clinical practice. For secondary prevention of CHD in women, the American Heart Association (AHA) recommends against initiating hormone replacement therapy (HRT) based on studies that have shown no benefit and early harm. For patients with CHD already on HRT, the decision to continue or stop HRT should be based on established noncoronary benefits and risks and patient preference. There are insufficient data to suggest that HRT should be initiated for the sole purpose of primary prevention of CHD. Because the new AHA guidelines recommend placing significant weight on the noncardiac benefits and risks of HRT, it is important to become familiar with these noncardiac effects. It is also important to understand the evidence supporting the AHA's decision to recommend placing relatively less weight on the cardiac effects of HRT. PMID- 11858776 TI - Clinical Acute Lung Injury and Acute Respiratory Distress Syndrome. AB - This article provides a description of the clinical disorders associated with the development of acute noncardiogenic pulmonary edema, better known as clinical acute lung injury (ALI) or the acute respiratory distress syndrome (ARDS). Much has been learned about the mechanisms by which the lung is injured in patients with sepsis, pneumonia, aspiration of gastric contents, and following major trauma. In the last 5 years, major progress has been made in the treatment of patients with ALI/ARDS. A lung protective ventilatory strategy with a low tidal volume (6 mL/kg/predicted body weight) in conjunction with a plateau pressure limit of 30 cm H(2)0 attenuated the severity of clinical lung injury and reduced mortality by 22%. Ironically, after years of searching for anti-inflammatory treatments for ALI/ARDS, it turns out that a lung protective ventilatory strategy has proven to be the most efficacious anti-inflammatory treatment ever discovered for ALI/ARDS. However, it is still possible that pharmacologic treatments also may enhance survival. For example, a recent report that activated protein C reduces mortality in patients with sepsis raises hope that the incidence and severity of sepsis-induced ALI/ARDS may be reduced by treatment with this agent that has both anti-inflammatory and anticoagulant properties. Also, therapy directed at hastening the resolution of lung injury by increasing the functional recovery of the alveolar epithelium may be of value, both in diminishing the fibroproliferative phase of ALI/ARDS as well as accelerating the resolution of alveolar edema. PMID- 11858778 TI - Respiratory and Hemodynamic Management After Cardiac Surgery. AB - In this new era of managed care, the emphasis has been on the reduction of intensive-care stay after coronary artery bypass surgery. "Fast-track" or rapid weaning protocols have become increasingly popular due to evidence that shows their cost-effectiveness and safety. With new advances in surgical and anesthetic techniques, the goal is often to have patients extubated within 4 to 6 hours upon arrival in the intensive-care unit. Patients who are not candidates for the fast track protocol are often those who either have poor respiratory function and a large A-a gradient or those who have hemodynamic instability from poor cardiac function after bypass. These patients need more intensive care and more traditional weaning from mechanical ventilation. Those that are not able to wean from the mechanical ventilator within a few weeks are candidates for tracheostomy in order to avoid complications from prolonged endotracheal intubation and to improve pulmonary toilet. The treatment of perioperative low cardiac output syndrome is another goal after bypass surgery. Poor cardiac function can be managed with a variety of vasopressor and inotropic agents based on what the suspected derangement is from clinical examination and hemodynamic measurements (eg, low preload, low cardiac index, high or low systemic vascular resistance). Another modality that has been shown to have benefit on reducing hospital stay and costs is prophylaxis for atrial fibrillation, which may occur in 40% of patients who undergo bypass surgery and in 60% of those who undergo valve replacement surgery. Beta-blockers and amiodarone have both been found to be effective as prophylaxis against postoperative atrial fibrillation. PMID- 11858777 TI - Cardiovascular Complications of Collagen Vascular Disease. AB - Collagen vascular diseases commonly affect the heart; cardiovascular events are the major cause of mortality in people with these diseases. A striking feature of the cardiac involvement in individuals with systemic lupus erythematosus (SLE) and rheumatoid arthritis is aggressive and accelerated atherosclerosis; women with SLE in the 35- to 44-year-old age group are more than 50 times more likely to suffer myocardial infarction than are matched controls. Traditional risk factors contribute to the accelerated atherosclerosis, but cannot explain the extent of risk. It is possible that the inflammatory process, which is similar to the inflammatory process in atherosclerosis, pays a critical pathophysiologic role. It is critically important to identify the presence of traditional cardiovascular risk factors (eg, tobacco usage, hypertension, hypercholesterolemia, diabetes, homocysteinemia), and to modify these to secondary prevention targets. Cardiac valvular disease is common in individuals with SLE and rheumatoid arthritis; its presence should be anticipated and subacute bacterial endocarditis prophylaxis precautions initiated. Cardiac autonomic neuropathy and conduction disturbances are common in people with heart disease related to systemic sclerosis and human leukocyte antigen B27; these patients should be monitored carefully for evidence of dysrhythmias. PMID- 11858779 TI - Cardiovascular Complications of Neuromuscular Disorders. AB - In the past decade, advances in molecular genetics have shown that many familial neuromuscular and cardiovascular diseases share a common pathophysiology. They are caused by inherited mutations in the cellular cytoskeleton of cardiac and skeletal muscle cells. The clinical manifestation of cardiac disease in neuromuscular disorders is common and their management should include both periodic cardiac assessment and appropriate symptomatic and definitive therapy. Dilated cardiomyopathy is a common complication of neuromuscular diseases. Cardiac function may decline progressively as part of the natural history of the disease, but current medical therapy, including angiotensin-converting enzyme inhibitors, beta-blockers, and diuretics, can be used to alleviate symptoms of left ventricular dysfunction. Conduction disturbances may be an important cause of mortality, especially in patients with Emery Dreifuss muscular dystrophy, Kearns-Sayre syndrome, and myotonic dystrophy, and thus pacemaker implantation can be life-saving. Rhythm disturbances, such as atrial fibrillation and ventricular tachyarrhythmias, have been reported in patients with neuromuscular diseases. Treatment is based on preventing sudden death and embolic phenomena and cardioverting or controlling atrial fibrillation. In these patients, problems may arise with anticoagulation and antiarrhythmic therapy due to the inherent locomotor instability associated with the disease, and the presence of concomitant atrioventricular disease. Although uncommon, hypertrophic cardiomyopathy may be a feature of some neuromuscular disorders. Patients should undergo regular risk stratification for sudden cardiac death and symptoms such as heart failure can be treated with medical therapy. PMID- 11858781 TI - [Bioterrorism and public health]. PMID- 11858780 TI - Endocrine Heart Disease. AB - With the increasing prevalence of obesity and diabetes mellitus in the United States, associated cardiovascular disease is reaching epidemic proportions with staggering economic and societal impact. Numerous studies have demonstrated the poorer prognosis associated with chronic coronary artery disease and acute coronary syndromes in patients with diabetes compared with nondiabetic patients. Although the therapeutic strategy is largely the same for the two populations, proper management of the diabetic patient with cardiovascular disease must account for the associated metabolic disturbances. Thyroid disease is the next most common endocrine disorder that affects proper function of cardiovascular patients; all patients presenting with coronary artery disease or cardiac arrhythmias should undergo screening with a sensitive thyroid-stimulating hormone assay and appropriate treatment when necessary. Though these areas are the most common points of intersection between the cardiologist and endocrinologist, a thorough understanding of the impacts of each endocrine system on cardiac function is essential to recognize disease entities that often present with a cardiovascular manifestation or affect patients with a primary cardiovascular disease. PMID- 11858782 TI - [Estimating the completeness of AIDS reporting in Spain]. AB - OBJECTIVE: To evaluate AIDS case reporting in the Spanish regions as compared with the AIDS deaths registered in mortality statistics, in order to identify possible deviations indicative of underreporting. METHODS: We carried out an ecological analysis taking each region as a unit. We compared incidence and mortality obtained from the AIDS reporting system with the HIV/AIDS deaths obtained from the mortality statistics of the Spanish Institute for Statistics. The 1986-1998 period was analysed globally, then the analysis was repeated for the 1995-1998 period. RESULTS: In the 1986-1998 period there was a good correlation (r = 0.93) between the AIDS incidence rates and HIV/AIDS mortality rates obtained from death statistics. Some regions presented an AIDS incidence lower than expected according to their mortality rate when it was compared with the national average, with Asturias (27%), Comunidad Valenciana (26%), Andalucia (20%), Ceuta (18%) and Cantabria (13%) standing out. Taking as a reference the five regions with the highest completeness, the underreporting in Spain was of 13%. In the 1995-1998 period these deviations increased moderately. In the 1986 1998 period, 18.9% less deaths were notified to the AIDS reporting system in comparison with death statistics, showing a great variability between regions. CONCLUSIONS: In Spain the reporting level of AIDS cases is acceptable for the aims of surveillance, although in some regions measures to improve it should be taken. The notification of deaths to the AIDS reporting system presents great deficiencies. PMID- 11858783 TI - [Validation of self-reported weight and height in an adolescent population]. AB - INTRODUCTION: The aim of this study was to examine the relationship between self reported population in order to evaluate the validity of self-reported measures for the purpose of estimating the prevalence of low (less-than-or-equal 15th percentile) and high (greater-than-or-equal 85th percentile) body mass index (BMI) in the study population. SUBJECTS AND METHOD: Information on self-reported and objective weight and height was obtained from a representative sample of 3,244 adolescents, aged 15-18 years, in secondary education schools in the Autonomous Community of Madrid. We calculated the mean relative error; the correlation between subjective and objective parameters, sensitivity, specificity and predictive value positive of low and high BMIs. RESULTS: The mean relative errors were as follows: weight: +0.07% for males versus and 0.79% for females; height, +0.51% for males versus +0.98% for females; BMI: 0.88% for males versus 2.63% for females. The correlation between self-reported and objective BMI was 0.87 for males and 0.90 for females. The prevalence of high BMI was underestimated by 34.1% and 34.4% of females white that of low BMI was overestimated by 10.7% of males and 14.8% of females. CONCLUSION: Analysis of BMI as a continuous variable, based on self-reported weight and height measurement data, entails a small margin of error. However, its use as a categorical variable involves a considerable underestimate of the prevalence of high BMI, and an smaller overestimate of the prevalence of low BMI. PMID- 11858784 TI - [Contribution of women and low-income households to the provision of informal home healthcare]. AB - OBJECTIVE: To determine gender and socioeconomic inequalities in the production of informal home health care. METHODS: Cross-sectional survey. The study population was formed by 17.907 individuals interviewed in 1994 in the Spanish sample of 1994 the European Community Household Panel. The number of caregivers in the sample is 1.305 individuals (7.3% of the total). Two outcome measures were analysed: the risk of being an informal caregiver and the risk of caring more than 28 h weekly. The independent variables were the household income level, occupation, gender, age and self-perceived health status. The odds ratio are obtained from logistic regression models. RESULTS: Women are more likely than men to provide care for someone (74.4%). Low-income individual without occupation bear the greatest probability of providing care more than 28 h weekly OR = 2.24 (1.45-3.47). Bad self-perceived health status in people with occupation is associated with being a full-time caregiver OR = 4.51 (1.48-13.68). Age increase the probability both of being an informal caregiver and to care more than 28 hours a week. CONCLUSION: Women, low-income social classes and aged people bear the greatest burden of giving care. PMID- 11858785 TI - [Risk factors associated to tuberculosis patients with positive sputum microscopy]. AB - OBJECTIVE: The main factor responsible for producing new cases of tuberculosis by exogen transmission is the existence of Mycobacterium tuberculosis in the sputum of tuberculosis patients. The objective of this study was to determine the risk factors associated to positive smear tuberculosis cases in Lleida (Spain). METHODS: We studied new cases of tuberculosis over the period 1992-1998. The variables considered were: age, gender, coinfection with HIV, injection drug use (IDU), consumption of alcohol, existence of caverns in thorax X-ray, and laboratory results of direct smear examination. We calculated the incidence rates for 100,000 persons-year. The association between the dependent variable case of tuberculosis with positive Mycobacterium tuberculosis smear and the remaining independent variables were assessed obtaining odds ratios (OR) and 95% confidence intervals (CI) (crude cOR and adjusted aOR by non conditional logistic regression). RESULTS: We detected 905 new cases of tuberculosis. The rates in the period 1992-1998 decreased from 38.8 to 30.8. 44.9% of cases (n = 406) presented a positive direct smear. There was a positive association between the risk of being a positive smear patient and having caverns in thorax x-ray (aOR = 6.8; 95% CI, 4.8-95), being male (aOR = 1.8; 95% CI, 1.3-2.6) and consuming alcohol (aOR = 1.6; 95% CI,1.1-2.3) and this risk was significant lower in those under 15 years old (aOR = 0.2; 95% CI, 0.1-0.5), and coinfection with HIV (aOR = 0.5; 95% CI, 0.3-0.9). CONCLUSIONS: HIV infection implies a reduced impact in endemecity of tuberculosis. Study of contacts and the implementation of directly observed treatment must be considered not only for coinfected HIV cases and IDU but also for adult males, especially those who consume alcohol, particularly when they have caverns. PMID- 11858786 TI - [Explanatory variables of avoidable cost produced by not prescribing a cheaper equivalent drug]. AB - OBJECTIVE: To identify the explanatory variables of avoidable cost produced by not prescribing a cheaper equivalent drug. METHODS: We conducted a cross sectional study of 1,500 primary care physicians in Galicia (northwest Spain). The sample consisted of 405 primary care physicians. The following independent variables were collected through a postal questionnaire physicians' training and specialty, physicians' perception of the quality of available drug information sources, type of practice, and number of patients. The environmental characteristics of the practice were obtained from secondary sources. Multiple regression models were constructed using as dependent variables two indicators based on unnecessary cost produced by not prescribing cheaper equivalents. RESULTS: The response rate was 77,4%. No association was found between prescription efficiency and the level of the physician's training, or with the variables of utilization or the quality of commercial and institutional information. The variable of health model was not significant. The only statistically significant variables were the number of identification cards and the percentage of pensioners' identification cards. CONCLUSIONS: The results of this study suggest that physicians attach little importance to prescription efficiency. Thus, to decrease avoidable cost produced by not prescribing a cheaper equivalent drug, the medical curriculum should be modified to ensure that is contents reflect national health priorities and current resources. PMID- 11858787 TI - Transformation and trends in preventive and social medicine education at the undergraduate level in a Brazilian medical school. AB - In the present study we discuss some transformations in undergraduate training in Preventive and Social Medicine in the Department of Social Medicine of the Faculty of Medicine of Ribeiro Preto, University of So Paulo, from 1993 to 1999. Aspects of the relationship between medical training and the reorganization of local services of the Brazilian national health system, and between graduate teaching in Preventive and Social Medicine and medical education as a whole are discussed. The crisis in Preventive and Social Medicine and its influence of medical training are evaluated. Trends for the application of a body of knowledge of the specialty and for the relationship between the department and the medical school are discussed. PMID- 11858789 TI - [The role of private insurance in public health care systems: conceptual framework and policies]. AB - The structure of the health care system ans specifically the type and amount of the public and private mix is not a closed issue. This article provides and update of the arguments that justify public intervention in health, and emphasizes the failures of the private insurance market that call for mandatory universal health insurance, although that does not necessarily mean that state has to be the insurer. The relationship between both sectors and the variables determining the relative level of expenditure in both are also analyzed. Following the literature on the public provision of private goods, the level of expenditure in a democracy is seen to depend on the preferences of the median voter, where private insurance usually tops up public insurance. The key variable determining the decision to buy additional private insurance is the difference in quality, defined broadly, between both sectors. Concerning policies, the appropriateness of fiscal incentives to promote the uptake of private insurance is discussed and it is concluded that there is no clear evidence of its suitability. Also, it is argued that models in which the public and private sectors appear totally segregated or totally integrated are preferable to intermediate models, in which both sectors appear combined. Medical coverage bought by an informed agent in exchange for a capitation payment seems a better way to integrate the private sector than through a system of vouchers. PMID- 11858788 TI - [Substitution indexes in ambulatory surgery: measure, count and compare]. AB - OBJECTIVE: Compare the substitution indexes of the ambulatory surgery activity done in the Vall d'Hebron Hospital with those indexes obtained from the all public hospitals of Catalonia which constitute the Public Hospital Net. METHODS: There have been calculated the substitution indexes of the 65% of DRGs which are treated with ambulatory surgery at the Vall d'Hebron Hospital and in the global of public hospital net in Catalonia. For this purpose, it has been used the MBDS of the years 1998 and 1999. The means of the substitution indexes have been compared with a Z test. RESULTS: The means of the substitution indexes are of a 48.88% for the Vall d'Hebron Hospital and of a 35.14% for the whole public hospitals. The difference is statically significant (p < 0.0001). Moreover, the highest substitution indexes are the same for both groups and are the follow: 039, 229,364, 262, 062, 362 y 342. CONCLUSIONS: Taking into account the present techniques of benchmarking in the health care field, this work shows that the substitution indexes are a valid tool that can be used to compare hospitals, specially in the case that it could be possible to work with robust data bases. PMID- 11858790 TI - [Access to therapeutic innovations in the least developed countries]. PMID- 11858791 TI - [Access to essential drugs in developing countries]. AB - Every year infectious diseases kill about 13 million people, about 30.000 deaths a day. Almost half of the victims are children younger than 5 years old, most of them belong to developing countries. Most of the premature deaths and the incapacity cases associated to infectious diseases could be avoided if the poor had access to medicines. In the developing world, the poverty of the families, the inappropried public expense and the lack of sanitary infrastructures get together to leave out of the reach of the poor the possibility of an adequate medical treatment. According to World Health Organization (WHO) in the developing countries about 2,000 million people lack access to essential medicines. The new patent rules of the World Trade Organization (WTO) will reduce even more access to essential medicines to the poor. Granting trade exclusivity rights to the owners of the patents for a period of at least 20 years limits the right of the governments to allow production, trade and importation of low cost copies (generics) of patented drugs. It is not a theoretical or far away menace. The application of these more strict rules has all ready caused serious problems to developing countries producers of generics as India or Brazil or to importers of these as South Africa or Kenya. They have been press, taken to court and threaten of sanctions by the pharmaceutical industry and some developed countries governments. The decision of some developing countries to fight for their most needed people's health added to the international campaign for the access to essential medicines defended by some non governmental organizations like Act Up, Treatment Action Campaign, Doctors Without Borders and Intermon Oxfam have implied small victories on behalf of the poor countries access to medicines. But deeper changes in the patent rules and the investigation of the diseases of the developing world are needed to improve health in the developing countries. PMID- 11858792 TI - [Intellectual property, patents and access to medication in developing countries]. AB - Pharmaceutical industry depends on continuing research and innovation to survive. This is a highly uncertain process due to the fact that only one in 20,000 screened molecules becomes a new and effective medicine. This process takes about 10 years to complete at a cost of e 480,000 per new compound. It is absolutely mandatory that intellectual property rights be protected in order to allow for investment of revenues in research and development of new medicines destined to cure or alleviate all types of disease, including those mainly afflicting the population in developing countries. The investigative drive of pharmaceutical companies has given the sector a leadership role in the search for global health. This is a responsibility the sector acknowledges and for that reason numerous laboratories are active partners of governments, private foundations, international organizations and N.G.O.s in a myriad of initiatives focused on alleviating most urgent health needs in developing countries. However, the true and final solution of this problem will only be accomplished when the social and political issues plagueing these societies are tackled. PMID- 11858793 TI - [Improving in the quality of mortality coding through press reports]. PMID- 11858794 TI - Ginseng and ginsenoside Rg3, a newly identified active ingredient of ginseng, modulate Ca2+ channel currents in rat sensory neurons. AB - There is increasing evidence that ginseng influences pain modulation. In spite of extensive behavior studies, the detailed mechanism of ginseng actions at the cellular level and the identity of the active substance have not been elucidated yet. Whole-cell patch-clamp recordings were used to examine the modulation of high-voltage-activated Ca2+ channel currents by ginseng total saponins and its various individual ginsenosides in rat dorsal root ganglion neurons. Application of ginseng total saponins suppressed Ca2+ channel currents in a dose-dependent manner. Occlusion experiments using selective blockers revealed that ginseng total saponins could modulate L-, N-, and P-type currents. The co-application of ginseng total saponins and the gamma-opioid receptor agonist, D-Ala(2), N MePhe(4), Gly(5)-ol-enkephalin (DAMGO), produced non-additive effects in most cells tested and each effect was significantly relieved by a depolarizing prepulse. Overnight treatment of cells with pertussis toxin profoundly reduced the inhibition. Furthermore, we now report that ginsenoside Rg3, among the major fractions of ginseng saponins, is a newly identified active component for the inhibition. These results suggest that the modulation of Ca2+ channels by ginseng total saponins, in particular by ginsenoside Rg3, could be part of the pharmacological basis of ginseng-mediated antinociception. PMID- 11858795 TI - Quinacrine increases endothelial nitric oxide release: role of superoxide anion. AB - The effect of acute quinacrine treatment on agonist-induced nitric oxide (NO) release was investigated in cultured human endothelial cells using electrochemical monitoring of the in situ NO concentration. Quinacrine dose dependently increased NO release with an apparent EC50 of 0.2 microM and a maximal effect at 1 microM. Quinacrine did not modify the dependence of NO release on extracellular L-arginine. Acceleration or deceleration of O2- dismutation, which altered NO release in control cells, did not modify it in quinacrine-treated cells. Quinacrine did not modify NO amperometric signal or reaction with O2- produced by xanthine oxidation. In the presence of quinacrine, agonist-induced NO release became Mg2+ -independent and could not be attributed to an inhibition of phospholipase A2 activity. Quinacrine made NO release insensitive to Cu2+ chelation. The present study demonstrates that acute treatment by low quinacrine concentrations increases endothelial NO release, possibly through an inhibition of O2- production. PMID- 11858796 TI - Increased expression of G protein-coupled receptor kinases in cystic fibrosis lung. AB - A reduction in airway beta-adrenoceptor density has been reported in cystic fibrosis lung but the mechanism underlying this defect remains unclear. In this study, we have investigated whether the decrease in beta2-adrenoceptor associates with altered G protein-coupled receptor kinase (GRK) levels. We assessed GRK activity by rhodopsin phosphorylation, and beta2-adrenoceptor and GRK at the mRNA and protein levels by Northern and Western blotting in peripheral lung samples from normal donors and patients with cystic fibrosis. GRK activity was significantly increased in peripheral cystic fibrosis lung with parallel increases in GRK2/5 mRNAs and protein expression. Functionally, isoproterenol stimulated adenylyl cyclase activity was also diminished by 65% in cystic fibrosis lung homogenates. These data suggest that the increase in GRK activity may be one of the mechanisms underlying alterations in the coupling between beta2 adrenoceptor and adenylyl cyclase via G-protein and may thus contribute to the downregulation of beta2-adrenoceptor in cystic fibrosis lung. PMID- 11858797 TI - Pharmacological characterisation of the goldfish somatostatin sst5 receptor. AB - Somatostatin (somatotropin release inhibiting factor, SRIF), exerts its effects via specific G protein coupled receptors of which five subtypes have been cloned (sst1-5). Recently, SRIF receptors have also been cloned from fish tissues. In this study, goldfish sst5 receptors (gfsst5) were expressed and characterised in the Chinese hamster lung fibroblast cell line, that harbours the luciferase reporter gene driven by the serum responsive element (CCL39-SRE-Luci). The agonist radioligands [125I]-LTT-SRIF-28 ([Leu8, DTrp22, 125I-Tyr25]SRIF-28) and [125I][Tyr10]cortistatin-14 labelled similar receptor densities with high affinity and in a saturable manner (pKd: 9.99-9.71; Bmax: 300-350 fmol mg-1). 5' Guanylyl-imidodiphosphate inhibited radioligand binding to some degree (38.5 57.9%). In competition binding studies, the pharmacological profile of SRIF binding sites defined with [125I]LTT-SRIF-28 and [125I][Tyr10]cortistatin-14 correlated significantly (r2=0.97, n=20). Pharmacological profiles of human and mouse sst5 receptors expressed in CCL39 cells correlated markedly less with those of the gfsst5 profile (r2=0.52-0.78, n > or = b16). Functional expression of the gfsst5 receptor was examined by measurement of agonist-induced luciferase expression and stimulation of [35S]GTPgammaS ([35S]guanosine 5'-O-(3 thiotriphosphate) binding. Profiles were similar to those achieved in radioligand binding studies (r2=0.81-0.93, n=20), although relative potency (pEC50) was reduced compared to pKd values. Relative efficacy profiles of luciferase expression and [35S]GTPgammaS binding, were rather divergent (r2=0.48, n=20) with peptides showing full agonism at one pathway and absence of agonism at the other. BIM 23056 (D-Phe-Phe-Tyr-D-Trp-Lys-Val-Phe-D-Nal-NH2) acted as an antagonist on the effects of SRIF-14 (pKB=6.74 +/- 0.23) on stimulation of [35S]GTPgammaS binding. Pertussis toxin abolished the effect of SRIF-14 on luciferase expression and [35S]GTPgammaS binding suggesting coupling of the receptor to G(i)/G(o) proteins. In summary, the present studies demonstrate that the gfsst5 receptor has a similar pharmacological profile and transductional properties to mammalian sst5 receptors. The difference in efficacy profiles defined using different functional assays suggests numerous, agonist specific, conformational receptor states, and/or ligand-dependent receptor trafficking. PMID- 11858798 TI - C- and N-terminal residue effect on peptide derivatives' antagonism toward the formyl-peptide receptor. AB - The biological action of several X-Phe-D-Leu-Phe-D-Leu-Z (X=3',5'-dimethylphenyl ureido; Z=Phe, Lys, Glu, Tyr) analogues was analysed on human neutrophils to evaluate their ability to antagonize formyl-peptide receptors. X-Phe-D-Leu-Phe-D Leu-Phe analogues obtained as C-terminal olo or amido derivatives and T-Phe-D-Leu Phe-D-Leu-Phe analogues (T=thiazolyl-ureido) were also analysed. The activities of pentapeptide derivatives were compared with those of X-Phe-D-Leu-Phe-D-Leu-Phe chosen as reference antagonist. Our results demonstrate that X-Phe-D-Leu-Phe-D Leu-Phe-olo, X-Phe-D-Leu-Phe-D-Leu-Glu and X-Phe-D-Leu-Phe-D-Leu-Tyr are more active antagonists than X-Phe-D-Leu-Phe-D-Leu-Phe. The presence of Lys (X-Phe-D Leu-Phe-D-Leu-Lys) seems, instead, to inhibit the formyl-peptide receptor antagonist properties. The presence of the N-terminal thiazolyl-ureido group seems to considerably contribute to the receptor antagonist properties of T-Phe-D Leu-Phe-D-Leu-Phe-OH. The introduction of the C-terminal methyl ester (T-Phe-D Leu-Phe-D-Leu-Phe-OMe) or amido group (X-Phe-D-Leu-Phe-D-Leu-Phe-NH2) appears detrimental for the affinity and formyl-peptide receptor antagonist properties of the Phe-D-Leu-Phe-D-Leu-Phe derivatives. The examined peptides inhibit superoxide anion production and lysozyme release more efficaciously than neutrophil chemotaxis. PMID- 11858799 TI - Noradrenergic lesions differentially alter the antidepressant-like effects of reboxetine in a modified forced swim test. AB - The novel antidepressant reboxetine is a selective norepinephrine reuptake inhibitor. In this study, the antidepressant-like effects of reboxetine were characterized in a modified rat forced swim test. Further, in order to investigate the role of the locus coeruleus and lateral tegmental noradrenergic systems in the mediation of reboxetine's effects, the impact of different chemical lesions of these two pathways was examined on the behavioral responses induced by reboxetine in the forced swim test. Reboxetine (5-20 mg/kg, s.c.) dose dependently decreased immobility and swimming behavior in the forced swim test while it simultaneously increased climbing behavior. These effects were similar to those previously demonstrated with tricyclic antidepressants and are indicative of reboxetine's effects on the noradrenergic system. Discrete local injections of the neurotoxin 6-hydroxydopamine were employed to lesion the ventral noradrenergic bundle arising from cells located in the lateral tegmentum. This resulting lesion completely prevented reboxetine (10 mg/kg, s.c.)-induced decreases in immobility and increases in climbing behavior, demonstrating that an intact ventral noradrenergic bundle is required for the manifestation of reboxetine-induced antidepressant-like behavior in the test. In contrast, lesions of the dorsal noradrenergic bundle which consists of neurons arising from the nucleus locus coereleus, were achieved by systemic pretreatment with the selective noradrenergic neurotoxin N-(2-chloroethyl)-N-2-bromobenzylamine (DSP-4; 50 mg/kg, i.p.). The ability of reboxetine (10 mg/kg, s.c.) to increase climbing and decrease immobility was augmented by DSP-4 pretreatment. Furthermore, neither lesions of the dorsal noradrenergic bundle nor the ventral noradrenergic bundle altered baseline immobility scores in the forced swim test. Taken together, these data suggest that forebrain regions innervated by these two distinct noradrenergic pathways exert opposing influences on the behavioral response to reboxetine in the rat forced swim test. PMID- 11858801 TI - Mesaconitine-induced relaxation in rat aorta: involvement of Ca2+ influx and nitric-oxide synthase in the endothelium. AB - Aconiti tuber, roots of aconite (Aconitum japonicum), is an oriental herbal medicine used for centuries in Japan and China to improve the health of persons with a weak constitution and poor metabolism. We investigated the effects of mesaconitine, one of the aconite alkaloids in Aconiti tuber, on the contraction and free intracellular Ca2+ concentration ([Ca2+]i) level in isolated rat thoracic aorta. Mesaconitine at 30 microM inhibited 3 microM phenylephrine induced contraction in the endothelium-intact, but not endothelium-denuded, aortic rings. The effect of mesaconitine was dependent on external Ca2+ concentrations. The relaxation induced by mesaconitine was abolished by N(omega) nitro-L-arginine methyl ester (0.1 mM, an inhibitor of nitric-oxide synthase), as well as the relaxation induced by acetylcholine. Acetylcholine induced relaxation in two phases in our conditions; the initial phase was transient and external Ca2+ -independent, and the second phase was sustained and external Ca2+ dependent. Treatment with 100 nM thapsigargin, which depleted intracellular Ca2+ stores, inhibited acetylcholine-induced, but not mesaconitine-induced, relaxation. Mesaconitine increased the [Ca2+]i level in endothelial cells by influx of Ca2+ from extracellular spaces. These findings suggest that mesaconitine-induced Ca2+ influx and activation of nitric-oxide synthase in endothelial cells and, thus, induced vasorelaxation in rat aorta. PMID- 11858800 TI - Effects of brain mineralocorticoid receptor blockade on blood pressure and renal functions in DOCA-salt hypertension. AB - In normotensive rats, we have previously demonstrated a role of brain mineralocorticoid receptors in blood pressure and renal function control. In the present study, the coordinate cardiovascular and renal effects of brain mineralocorticoid receptor blockade were examined by intracerebroventricular (i.c.v.) administration of a selective mineralocorticoid receptor antagonist (RU28318; 3,3-oxo-7 propyl-17-hydroxy-androstan-4-en-17yl-propionic acid lactone) in rats with hypertension induced by deoxycorticosterone acetate (DOCA) and salt. DOCA pellets were implanted s.c. in male Wistar rats given 0.9% NaCl as drinking solution 3 or 5 weeks before assessment of the effects of i.c.v. injection of RU28318 on cardiovascular and renal functions. Changes in expression of brain angiotensinogen, atrial natriuretic peptide (ANP) and mineralocorticoid receptor mRNA in specific brain areas in 3-week DOCA-salt rats were evaluated by in situ hybridization. The rise in systolic blood pressure induced by DOCA-salt treatment was most marked during the first 3 weeks. At 3 and 5 weeks after implantation of the DOCA-pellets a single i.c.v. injection of 10 ng of RU28318 significantly decreased systolic blood pressure during 24 h as assessed at 2, 8 and 24 h, while heart rate was not altered. Increased urinary excretion of water and electrolytes was observed in 3- and 5-week DOCA-salt rats during the period 0-8 h after i.c.v. injection of RU28318 while the suppressed plasma renin activity was not affected. The expression of brain angiotensinogen, ANP and mineralocorticoid receptor mRNA was not altered by 3-week DOCA-salt treatment, but 3 h after i.c.v. injection of RU28318, mineralocorticoid receptor mRNA expression in hippocampal cell fields responded with an increase of about 40%. In conclusion, these results demonstrate that in rats with hypertension induced by DOCA-salt, brain mineralocorticoid receptor blockade affects renal function and blood pressure regulation. PMID- 11858802 TI - Effects of SQ 22536, an adenylyl cyclase inhibitor, on isoproterenol-induced cyclic AMP elevation and relaxation in newborn ovine pulmonary veins. AB - The effects of inhibition of adenylyl cyclase on isoproterenol-induced relaxation were determined in isolated pulmonary veins of newborn lambs (7-12 days old). In veins constricted with endothelin-1, isoproterenol at concentrations < or = 3 x 10(-9) M had no effect on the cyclic AMP (cAMP) content but caused up to 56% relaxation. At higher concentrations (> or = 10(-8) M), isoproterenol elevated cAMP content and caused further relaxation. In veins constricted with endothelin 1 or U46619 (9,11-dideoxy-11, 9-epoxymethanoprostaglandin prostaglandin F2alpha), the cAMP elevation but not relaxation caused by isoproterenol was abolished by SQ 22536 [9-(tetrahydro-2-furanyl)-9H-purin-6-amine; an adenylyl cyclase inhibitor]. The effects of isoproterenol on vessel tension and cAMP content were inhibited by propranolol. Rp-8-CPT-cAMPS [8-(4-Chlorophenylthio)-adenosine-3',5'-cyclic monophosphorothioate, Rp-isomer] and Rp-8-Br-PET-cGMPS [beta-phenyl-1, N2-etheno 8-bromoguanosine-3',5'-cyclic monophosphorothioate, Rp-isomer], inhibitors of cAMP- and guanosine-3',5'-cyclic monophosphate (cGMP)-dependent protein kinases, respectively, attenuated relaxation caused by a cAMP analog but not that by isoproterenol. In the crude membrane preparations of pulmonary veins, an increase in the activity of adenylyl cyclase caused by isoproterenol was abolished by propranolol and SQ 22536. These results suggest that cAMP may not play a critical role in isoproterenol-induced relaxation of pulmonary veins of newborn lambs. PMID- 11858803 TI - Inhibitory effects of azelastine on substance P-induced itch-associated response in mice. AB - The anti-pruritic mechanisms of azelastine were studied in mice. Scratching induced by intradermal histamine was inhibited by azelastine (30 mg/kg) and chlorpheniramine (30 mg/kg). Substance P-induced scratching was dose dependently suppressed by azelastine (3-30 mg/kg), but not by chlorpheniramine (10 and 30 mg/kg). Azelastine (30 mg/kg) inhibited the substance P-induced production of leukotriene B4, but not prostaglandin E2, in the skin. Azelastine (3-30 mg/kg) suppressed scratching induced by intradermal injection of leukotriene B4. The results suggest that inhibition of the production and action of leukotriene B4, as well as an anti-histamine action, is involved in the anti-pruritic action of azelastine. PMID- 11858804 TI - Cytokine modulation of granulocyte macrophage-CSF and granulocyte-CSF release from stimulated vascular smooth muscle cells. AB - Cytokine-stimulated vascular smooth muscle cells release the colony-stimulating factors (CSFs) granulocyte macrophage-CSF and granulocyte-CSF. We have investigated the effects of a range of cytokines on the release of CSFs from human vascular smooth muscle cells stimulated with interleukin-1beta. Interleukin 4 suppressed granulocyte macrophage-CSF release but potentiated granulocyte-CSF release; interferon-gamma inhibited the release of both, whilst interleukin-5 had no effect. Both interleukin-10 and interleukin-13 inhibited granulocyte macrophage-CSF release but did not affect granulocyte-CSF release. The ability of individual cytokines to differentially modulate CSFs has profound consequences for the populations of leukocytes present at the site of inflammation. PMID- 11858805 TI - Rabbit tendon cells produce MMP-3 in response to fluid flow without significant calcium transients. AB - Forces applied to tendon during movement cause cellular deformation, as well as fluid movement. The goal of this study was to test the hypothesis that rabbit tendon fibroblasts detect and respond to fluid-induced shear stress. Cells were isolated from the paratenon of the rabbit Achilles tendon and then subjected to fluid flow at 1 dyn/cm(2) for 6h in a specially designed multi-slide flow device. The application of fluid flow led to an increased expression of the collagenase-1 (MMP-1), stromelysin-1 (MMP-3), cyclooxygenase II (COX-2) and interleukin-1beta (IL-1beta) genes. The release of proMMP-3 into the medium exhibited a dose response with the level of fluid shear stress. However, not all cells aligned in the direction of flow. In other experiments, the same cells were incubated with the calcium-reactive dye FURA-2 AM, then subjected to laminar fluid flow in a parallel plate flow chamber. The cells did not significantly increase intracellular calcium concentration when exposed to fluid shear stress levels of up to 25 dyn/cm(2). These results show that gene expression in rabbit tendon cells is sensitive to fluid flow, but that signal transduction is not dependent on intracellular calcium transients. The upregulation of the MMP-1, MMP-3 and COX 2 genes shows that fluid flow could be an important mechanical stimulus for tendon remodelling or injury. PMID- 11858806 TI - Residual stress due to curing can initiate damage in porous bone cement: experimental and theoretical evidence. AB - Residual stress due to shrinkage of polymethylmethacrylate bone cement after polymerisation is possibly one factor capable of initiating cracks in the mantle of cemented hip replacements. No relationship between residual stress and observed cracking of cement has yet been demonstrated. To investigate if any relationship exists, a physical model has been developed which allows direct observation of damage in the cement layer on the femoral side of total hip replacement. The model contains medial and lateral cement layers between a bony surface and a metal stem; the tubular nature of the cement mantle is ignored. Five specimens were prepared and examined for cracking using manual tracing of stained cracks, observed by transmission microscopy; cracks were located and measured using image analysis. A mathematical approach for the prediction of residual stress due to shrinkage was developed which uses the thermal history of the material to predict when stress-locking occurs, and estimates subsequent thermal stress. The residual stress distribution of the cement layer in the physical model was then calculated using finite element analysis. Results show maximum tensile stresses normal to the observed crack directions, suggesting a link between residual stress and pre-load cracking. The residual stress predicted depends strongly on the definition of the reference temperature for stress locking. The highest residual stresses (4-7 MPa) are predicted for shrinkage from maximum temperature; in this case, magnitudes are sufficiently high to initiate cracks when the influence of stress raisers such as pores or interdigitation at the bone/cement interface are taken into account (up to 24 MPa when calculating stress around a pore according to the method of Harrigan and Harris (J. Biomech. 24(11) (1991) 1047-1058). We conclude that the damage accumulation failure scenario begins before weight-bearing due to cracking induced by residual stress around pores or stress raisers. PMID- 11858807 TI - An elasto-plastic finite element model for polyethylene wear in total hip arthroplasty. AB - A new finite element model (FEM) based on an elasto-plastic behavior of ultra high molecular weight polyethylene (UHMWPE) was used to study the wear behavior of UHMWPE acetabular cup, which has a 32 mm diameter femoral head. The model imposed a plastic yield stress of 8 MPa on the UHMWPE so that any stresses beyond this would automatically be redistributed to its neighbor. The FEM model adopted a unique mesh design based on an open cube concept which eliminated the problems of singularities. Wear prediction combined the influences of contact stress, sliding distance and a surface wear coefficient. The new model predicted significantly higher volumetric wear rate (57 mm(3)/yr) well within the average reported clinical values. The model was also used to study the effect of friction and clearance between the acetabular cup and the femoral head. Increase in friction increased the volumetric wear rate but did not appear to affect the linear wear rate, which remained at 0.12 +/- 0.02 mm/yr. The predicted wear was sensitive to clearance. It was found that when the clearance was close to 0 and >0.5mm, severe wear occurred. The best clearance range was between 0.1 and 0.15 mm where the average linear wear rate was 0.1mm/yr and the volumetric wear was 55 mm(3)/yr. The present work indicates the importance of avoiding too tight or too loose a diametrical clearance. PMID- 11858808 TI - Interfragmentary surface area as an index of comminution energy: proof of concept in a bone fracture surrogate. AB - Fracture mechanics theory postulates a monotonic relationship between energy absorption and fracture surface generation. We hypothesized that this relationship was demonstrable to the point that, on a continuous scale, comminuted fractures created with disparate levels of energy delivery could be discriminated. Using a bone fracture surrogate in conjunction with digital image analysis of CT fracture data, we measured the surface area freed by controlled, discrete fracture simulations. Prior to these simulations, the reproducibility of the digital image analysis algorithm was validated with repeated measurements by two different operators. The parametric fracture series results showed a statistically significant difference in measured de novo surface area between four specimen groups, over a range of input energies from 1.4 x 10(10)-9.1 x 10(10)J/m(3) (or 12.5-80.2J/specimen). The results of this study provide confirmation that comminution severity can indeed be measured on a continuous scale, based on energy absorption (another clinically meaningful index). PMID- 11858809 TI - Mechanically simulated muscle forces strongly stabilize intact and injured upper cervical spine specimens. AB - Although muscles are assumed to be capable of stabilizing the spinal column in vivo, they have only rarely been simulated in vitro. Their effect might be of particular importance in unstable segments. The present study therefore tests the hypothesis that mechanically simulated muscle forces stabilize intact and injured cervical spine specimens. In the first step, six human occipito-cervical spine specimens were loaded intact in a spine tester with pure moments in lateral bending (+/- 1.5 N m), flexion-extension (+/- 1.5 N m) and axial rotation (+/- 0.5 N m). In the second step, identical flexibility tests were carried out during constant traction of three mechanically simulated muscle pairs: splenius capitits (5 N), semispinalis capitis (5 N) and longus colli (15 N). Both steps were repeated after unilateral and bilateral transection of the alar ligaments. The muscle forces strongly stabilized C0-C2 in all loading and injury states. This was most obvious in axial rotation, where a reduction of range of motion (ROM) and neutral zone to <50% (without muscles=100%) was observed. With increasing injury the normalized ROM (intact condition=100%) increased with and without muscles approximately to the same extend. With bilateral injury this increase was 125-132% in lateral bending, 112%-119% in flexion-extension and 103-116% in axial rotation. Mechanically simulated cervical spine muscles strongly stabilized intact and injured cervical spine specimens. Nevertheless, it could be shown that in vitro flexibility tests without muscle force simulation do not necessarily lead to an overestimation of spinal instability if the results are normalized to the intact state. PMID- 11858810 TI - Fatigue damage of human tendons. AB - The study was designed to examine the effects of partial fatigue on specific mechanical parameters which characterise human tendons in vitro. Specimens prepared from 12 intact Extensor digitorum longus tendons of the foot were subjected to partial fatigue, equivalent to 25% of the median fatigue life, by a cyclic square tension-tension stress waveform at the physiological frequency of 4 Hz. The maximum stress was set at a value corresponding to 20% of the calculated ultimate tensile strength (UTS) of 100 MPa. The minimum stress was set at 1% of the UTS. Dynamic characterisation was performed at stress levels of 10% and 20% of the UTS prior to and following partial mechanical fatigue. Subsequent quasi static tests were performed on some of the specimens. Comparative analysis of the damage ratios (DRs) of quasi-static and dynamic mechanical parameters suggested the use of the DR for dynamic tensile modulus as a good indicator of damage inflicted by mechanical fatigue. Such an approach might be used for an in vivo assessment of tendon damage. PMID- 11858811 TI - Changing the structurally effective mineral content of bone with in vitro fluoride treatment. AB - Bovine femur cortical bone specimens were tested in tension after being treated in vitro for 3 days with sodium fluoride solutions of different molarity (0.145, 0.5, and 2.0M). The treatments alter the mechanical properties of the bone samples with different degrees as compared to control samples (untreated). The mechanical properties of the treated samples have lower elastic modulus, yield and ultimate stress, acoustic impedance and hardness, and higher ultimate strain and toughness as compared to control samples. The observed effects were intensified with the increasing molarity of the treatment solutions. This study shows that the fluoride treatment can be used to investigate the composite behavior of bone tissue by altering the structurally important bone mineral content in a controlled manner. PMID- 11858812 TI - Quantification of a rat tail vertebra model for trabecular bone adaptation studies. AB - A feedback controlled loading apparatus for the rat tail vertebra was developed to deliver precise mechanical loads to the eighth caudal vertebra (C8) via pins inserted into adjacent vertebrae. Cortical bone strains were recorded using strain gages while subjecting the C8 in four cadaveric rats to mechanical loads ranging from 25 to 100 N at 1 Hz with a sinusoidal waveform. Finite element (FE) models, based on micro computed tomography, were constructed for all four C8 for calculations of cortical and trabecular bone tissue strains. The cortical bone strains predicted by FE models agreed with strain gage measurements, thus validating the FE models. The average measured cortical bone strain during 25-100 N loading was between 298 +/- 105 and 1210 +/- 297 microstrain (muepsilon). The models predicted average trabecular bone tissue strains ranging between 135 +/- 35 and 538 +/- 138 mu epsilon in the proximal region, 77 +/- 23-307 +/- 91 muepsilon in the central region, and 155 +/- 36-621 +/- 143 muepsilon in the distal region for 25-100 N loading range. Although these average strains were compressive, it is also interesting that the trabecular bone tissue strain can range from compressive to tensile strains (-1994 to 380 mu epsilon for a 100 N load). With this novel approach that combines an animal model with computational techniques, it could be possible to establish a quantitative relationship between the microscopic stress/strain environment in trabecular bone tissue, and the biosynthetic response and gene expression of bone cells, thereby study bone adaptation. PMID- 11858813 TI - Effect of normalization and phase angle calculations on continuous relative phase. AB - The purpose of this investigation was to determine if phase plot normalization and phase angle definitions would have an affect on continuous relative phase calculations. A subject ran on a treadmill while sagittal plane kinematic data were collected with a high-speed (180 Hz) camera. Segmental angular displacements and velocities were used to create phase plots, and examine the coordination between the leg and thigh. Continuous relative phase was calculated with a combination of two different amplitude normalization techniques, and two different phase angle definitions. Differences between the techniques were noted with a root mean square (RMS) calculation. RMS values indicated that there were differences in the configuration of the non-normalized and normalized continuous relative phase curves. Graphically and numerically, it was noted that normalization tended to modify the continuous relative phase curve configuration. Differences in continuous relative phase curves were due to a loss in the aspect ratio of the phase plot during normalization. Normalization tended to neglect the nonlinear forces acting on the system since it did not maintain the aspect ratio of the phase plot. Normalization is not necessary because the arc tangent function accounts for differences in amplitudes between the segments. RMS values indicated that there were profound differences in the continuous relative phase curve when the phase angle was normalized and a phase angle was calculated relative to the right horizontal axis. PMID- 11858814 TI - Simulation of bone adaptive remodeling using a stochastic process as loading history. AB - In this simulation study for bone adaptive remodeling, loading conditions are described as stochastic processes to catch the unpredictable characteristics of daily physical activities, which are observed to be closely related with bone adaptive remodeling. This will not only eliminate the necessity of arbitrary choices for loading conditions, but also generate greater flexibility for simulations of bone adaptive remodeling. The sensitivity of simulation outcomes to the parameters in the simulation algorithm was examined by applying stochastic loading conditions on finite element models of simplified spine structures. In this way, the limitations induced by simplifying loading conditions into constant or cyclic loads can be avoided and, potentially, more clinical observations could be accommodated when more comprehensive finite element models are available. PMID- 11858815 TI - Measurement of canal occlusion during the thoracolumbar burst fracture process. AB - Post-injury CT scans are often used following burst fracture trauma as an indication for decompressive surgery. Literature suggests, however, that there is little correlation between the observed fragment position and the level of neurological injury or recovery. Several studies have aimed to establish the processes that occur during the fracture using indirect methods such as pressure measurements and pre/post impact CT scans. The purpose of this study was to develop a direct method of measuring spinal canal occlusion during a simulated burst fracture by using a high-speed video technique. The fractures were produced by dropping a mass from a measured height onto three-vertebra bovine specimens in a custom-built rig. The specimens were constrained to deform only in the impact direction such that pure compression fractures were generated. The spinal cord was removed prior to testing and the video system set up to film the inside of the spinal canal during the impact. A second camera was used to film the outside of the specimen to observe possible buckling during impact. The video images were analysed to determine how the cross-sectional area of the spinal canal changed during the event. The images clearly showed a fragment of bone being projected from the vertebral body into the spinal canal and recoiling to the final resting position. To validate the results, CT scans were taken pre- and post-impact and the percentage canal occlusion was calculated. There was good agreement between the final canal occlusion measured from the video images and the CT scans. PMID- 11858816 TI - Comments on the letter "Accelerometer and mechanomyogram". PMID- 11858818 TI - Stepping back in sprinting. PMID- 11858820 TI - Structure and function of major histocompatibility complex (MHC) class I specific receptors expressed on human natural killer (NK) cells. AB - Natural killer (NK) cells express receptors that are specific for MHC class I molecules. These receptors play a crucial role in regulating the lytic and cytokine expression capabilities of NK cells. In humans, three distinct families of genes have been defined that encode for receptors of HLA class I molecules. The first family identified consists of type I transmembrane molecules belonging to the immunoglobulin (Ig) superfamily and are called killer cell Ig-like receptors (KIR). A second group of receptors belonging to the Ig superfamily, named ILT (for immunoglobulin like transcripts), has more recently been described. ILTs are expressed mainly on B, T and myeloid cells, but some members of this group are also expressed on NK cells. They are also referred to as LIRs (for leukocyte Ig-like receptor) and MIRs (for macrophage Ig-like receptor). The ligands for the KIR and some of the ILT receptors include classical (class Ia) HLA class I molecules, as well as the nonclassical (class Ib) HLA-G molecule. The third family of HLA class I receptors are C-type lectin family members and are composed of heterodimers of CD94 covalently associated with a member of the NKG2 family of molecules. The ligand for most members is the nonclassical class I molecule HLA-E. NKG2D, a member of the NKG2 family, is expressed as a homodimer, along with the adaptor molecule DAP10. The ligands of NKG2D include the human class I like molecules MICA and MICB, and the recently described ULBPs. Each of these three families of receptors has individual members that can recognize identical or similar ligands yet signal for activation or inhibition of cellular functions. This dichotomy correlates with particular structural features present in the transmembrane and intracytoplasmic portions of these molecules. In this review we will discuss the molecular structure, specificity, cellular expression patterns, and function of these HLA class I receptors, as well as the chromosomal location and genetic organization. PMID- 11858821 TI - Evidences of conformational changes in class II Major Histocompatibility Complex molecules that affect the immunogenicity. AB - The N-terminal part of class II-associated invariant chain peptide (CLIP) is assumed to interact with an accessory peptide-binding site on the class II Major Histocompatibility Complex (MHC) molecule, and promote a conformational modification. We have linked this immunoregulatory segment (residues 81-88) to the N-terminus of the influenza hemagglutinin (HA) 307-319 epitope in order to evaluate relationships between the MHC conformational changes and their implication in immune responses. Our chimeric peptide, named CLIP-HA, bind with the same affinity to class II HLA-DR1 molecules as the HA peptide, and is normally recognized by HA-specific T cells. Interestingly, the presence of the N terminal CLIP region enhances the rate of association to soluble DR1 molecules but prevents the formation of SDS-resistant complexes. These features suggest the existence of HLA-DR1 conformational changes induced by the chimeric peptide. Furthermore, while in vitro HA and CLIP-HA peptides associated to DR1 could not be differentiated based on T-cell recognition, in vivo the CLIP residues strongly impaired the immunogenicity of HA epitope as assessed in HLA-DR1 transgenic mice. Our study demonstrates for the first time that MHC conformational changes, revealed at molecular level, may influence the immunogenicity. PMID- 11858822 TI - Neuroinvasion by pathogens: a key role of the complement system. AB - Complement (C) is one of the most critical defence mechanisms of the innate immunity against cerebral infection by viruses, bacteria and fungi, with different molecular pathways contributing to the clearance of the invading pathogens. There is now compelling evidence that C proteins can be synthesized by brain cells in response to the infectious challenge and leading to cytotoxic and cytolytic activities against the harmful intruders. However, since there is also emerging evidence that uncontrolled C biosynthesis/activation can lead to brain inflammation with loss of neurons and oligodendrocytes, it is important to highlight that C may have adverse effects in infectious diseases of the CNS and induce profound tissue damage. The role of C in brain infection may even be more versatile. Many invading pathogens are not helpless against C attack and can use the membrane-bound C molecules to invade the host, either by binding directly or after decoration with C fragments. During budding viruses can acquire complement inhibitors from the host cell membrane and thus behave like 'Trojan horses' that are sheltered from the local innate immune response. Moreover, pathogens have evolved means of molecular mimicry with the expression of C inhibitor-like molecules to escape recognition and clearance by the C system. We herein provide a comprehensive and insightful review of the expression and the role of the C system in the brain. The three main focuses are: (i) C activation and lysis of pathogens in the brain; (ii) C-dependent neuroinvasion mechanisms (iii) uncontrolled C activation in inflamed CNS contributing to tissue damage. PMID- 11858823 TI - Modeling MHC class II molecules and their bound peptides as expressed at the cell surface. AB - A detailed insight to the structure of a given major histocompatibility complex (MHC)-peptide complex can strongly support and also improve the analysis of the peptide binding capabilities of the MHC molecule and the characterization of the developing T cell response. The number of MHC class II-peptide crystal structures is limited, therefore constructing and analyzing computer models can serve as efficient complementary tools when someone deals with experimentally determined binding and/or functional data. Commercial programs are available for modeling protein and protein-protein complexes, in general. However, more accurate results can be obtained if the parameters are directly optimized to a given complex, especially in the case of special proteins as MHC class II, an integral membrane protein, whose functional parts behave like regular globular proteins. Here, we present the optimization of an approach used for modeling MHC class II molecules complexed with various peptides fitting into the binding groove and several ways to analyze them with the help of experimental data. PMID- 11858825 TI - Defective proliferative responses in B lymphocytes and thymocytes that lack neurofibromin. AB - Nf1(-/-) fetal liver cells were used to reconstitute B and T cells in Rag-1(-/-) mice. Lymphocyte development was largely unimpaired in the absence of neurofibromin. However antigen-receptor induced proliferation was defective in neurofibromin deficient peripheral B cells and CD4(+) single positive thymocytes. In contrast to its role as a negative regulator of proliferation in many other cell types, neurofibromin may be a positive regulator of lymphocyte proliferation. Peripheral B cells exhibited circumscribed defects in anti-IgM induced protein tyrosine phosphorylation, which may contribute to the unexpected proliferative defect seen in these cells. PMID- 11858824 TI - Molecular assembly of CD46 with CD9, alpha3-beta1 integrin and protein tyrosine phosphatase SHP-1 in human macrophages through differentiation by GM-CSF. AB - Human CD46, formerly membrane cofactor protein (MCP), binds and inactivates complement C3b and serves as a receptor for measles virus (MV), thereby protecting cells from homologous complement and sustaining systemic viral infection. CD46 on activated macrophages (Mphi) but not intact monocytes is presumed to be the factor responsible for virus-mediated immune modulation including down-regulation of IL-12 production. As CD46 is expressed on both Mphi and monocytes, the molecular mechanisms responsible for these distinct immune responses remain largely unknown. Here, we found that peripheral blood monocytes treated for 5--8 days with GM-CSF (i.e. mature Mphi) acquired the capacity to assemble CD9, alpha3-beta1 integrin and the tyrosine phosphatase SHP-1 with their CD46. Prior to this maturation stage, Mphi expressed sufficient amounts of CD9 and CD46 but showed no such complex formation, and as in intact monocytes MV replication was markedly suppressed. By flow cytometry and confocal microscopy, the complex was found to assemble on the surface in cells treated with approximately 6 days with GM-CSF but not for approximately 2 days. Notably, an alternative MV receptor SLAM CDw150 was neither expressed nor recruited to this complex throughout GM-CSF-mediated Mphi differentiation. These responses and molecular links were not reproduced in the hamster cell line CHO expressing human CD46 although these cells acquired high susceptibility to MV. Based on these observations, MV susceptibility in human myeloid lineages appears not to be as simple as that observed in human CD46-transfected non-myeloid cells. The molecular complex involving CD46 may confer high MV permissiveness leading to immune modulation in Mphi. PMID- 11858827 TI - Receptor-ligand interaction demonstrated in Brassica self-incompatibility. PMID- 11858826 TI - Sry and sex determination: how lazy can it be? PMID- 11858828 TI - QTL for timing: a natural diversity of clock genes. PMID- 11858829 TI - Five years of vector service for gene therapy. PMID- 11858835 TI - Splicing regulation as a potential genetic modifier. AB - Inherited diseases are associated with profound phenotypic variability, which is affected strongly by genetic modifiers. The splicing machinery could be one such modifying system, through a mechanism involving splicing motifs and their interaction with a complex repertoire of splicing factors. Mutations in splicing motifs and changes in levels of splicing factors can result in different splicing patterns. Changes in the level of normal transcripts or in the relative pattern of different mRNA isoforms affect disease expression, leading to phenotypic variability. Here, we discuss the splicing machinery in terms of its significance in disease severity and its potential role as a genetic modifier. PMID- 11858836 TI - The evolution of morphological complexity in zebrafish stripes. AB - The zebrafish pigment stripe pattern is a complex tissue containing iridophores, xanthophores and multiple melanocyte types. Mutational analysis reveals that both ancient and recent gene duplications are involved in the generation or maintenance of the pattern complexity. Receptor tyrosine kinases kit and fms, products of an ancient gene duplication, are required in distinct types of melanocytes and xanthophores. Transcription factors mitfa and mitfb, results of a teleost-specific duplication, partition gene expression and function between different sets of melanocytes. Understanding the roles of these duplicated genes in zebrafish allows us to predict roles for their precursors in ancestral vertebrates. PMID- 11858837 TI - Establishment of polarity in angiosperm lateral organs. AB - In seed plants, lateral organs such as leaves and floral organs are formed from the flanks of apical meristems. Therefore, they have an inherent positional relationship: organ primordia have an adaxial side next to the meristem, and an abaxial one away from the meristem. Surgical and genetic studies suggest that a morphogenetic gradient, which originates in the meristem, converts the inherent polarity into a functional one. Once an adaxial-abaxial axis of polarity is established within organ primordia, it provides cues for proper lamina growth and asymmetrical development. Several key participants in this process have been identified, and analyses of these genes support and refine our views of axis formation in plants. The complex relationships between and within various members of these plant-specific gene families (class III HD-ZIPs, YABBYs and KANADIs) might account for a substantial part of the morphological variation in lateral organs of seed plants. PMID- 11858838 TI - Genetic analysis of the mammalian cell death machinery. AB - Programmed cell death is used by multicellular organisms to eliminate excess, damaged or harmful cells. This process of cell suicide, defined in morphological terms as apoptosis, is crucial for developmental morphogenesis, tissue homeostasis and defense against pathogens. Over the past decade, our understanding of the genetic basis of the cell death machinery has grown exponentially using genetically modified organisms. In particular, inactivation of genes involved in cell death using homologous recombination in mice has provided an invaluable tool to understand the mechanisms, as well as the structural and functional consequences, of programmed cell death in mammals. This review discusses recent insights into the cellular death program as revealed by these mutant animals. PMID- 11858839 TI - A PUF family portrait: 3'UTR regulation as a way of life. AB - In eukaryotic cells, mRNAs are exquisitely controlled, often through regulatory elements in their 3' untranslated regions (3'UTRs). Proteins that bind to those sites are key players in controlling mRNA stability, translation and localization. One family of regulatory proteins--the PUF proteins--are not only structurally related, but also bind to 3'UTRs and modulate mRNA expression in a wide variety of eukaryotic species. They do so either by enhancing turnover or repressing translation, and act combinatorially with other regulatory proteins. Here, we discuss the evolution, biological function and mechanisms of action of the PUF protein family, and suggest that a primordial function of PUF proteins is to sustain mitotic proliferation of stem cells. PMID- 11858840 TI - SHOT: a web server for the construction of genome phylogenies. AB - With the increasing availability of genome sequences, new methods are being proposed that exploit information from complete genomes to classify species in a phylogeny. Here we present SHOT, a web server for the classification of genomes on the basis of shared gene content or the conservation of gene order that reflects the dominant, phylogenetic signal in these genomic properties. In general, the genome trees are consistent with classical gene-based phylogenies, although some interesting exceptions indicate massive horizontal gene transfer. SHOT is a useful tool for analysing the tree of life from a genomic point of view. It is available at http://www.Bork.EMBL-Heidelberg.de/SHOT. PMID- 11858843 TI - Synthesis and cholinesterase activity of phenylcarbamates related to Rivastigmine, a therapeutic agent for Alzheimer's disease. AB - In order to develop new cholinesterase agents effective against Alzheimer's disease (AD) we synthesized some phenylcarbamates structurally related to Rivastigmine and evaluated their in vitro and in vivo biological activity. Among the compounds which displayed the most significant in vitro activity, 1-[1-(3 dimethylcarbamoyloxyphenyl)ethyl]piperidine (31b), in addition to a simple and cheaper synthesis, showed lower toxicity and very similar therapeutic index in comparison with Rivastigmine. PMID- 11858845 TI - Quantitative structure--activity relationships of antimutagenic benzalacetones and 1,1,1-trifluoro-4-phenyl-3-buten-2-ones. AB - The antimutagenic activities (IC(50)) of benzalacetones (BZ) and 1,1,1-trifluo-4 phenyl-3-buten-2-ones (TF) against UV-induced mutagenesis in Escherichia coli WP2s(uvrA trpE) were quantitatively analyzed in terms of physicochemical parameters by regression analyses. Structural requirements for maximal potency were derived from the results of quantitative structure--activity relationship (QSAR) analyses: (1) ring substituents should be electron-withdrawing; (2) 2-OH substituents incapable of intramolecular hydrogen-bonding notably increase the potency; and (3) replacement of CH(3) group by CF(3) in the side chain enhances the activity. Contrary to our expectations, the best correlation lacked hydrophobic effects. Antimutagenic activities against gamma-induced mutagenesis in Salmonella typhimurium TA2638 were also studied for some derivatives in the BZ series, where, in addition to electronic and hydrogen-bonding factors, a hydrophobic term was also significant. Physicochemical meanings of the derived correlations are discussed. PMID- 11858844 TI - CoMFA analysis of the human beta(1)-adrenoceptor binding affinity of a series of phenoxypropanolamines. AB - A series of 36 phenoxypropanolamines was examined to determine the structure- activity relationships of beta-adrenoceptor (beta-AR) antagonists for the human beta(1)-AR. The binding affinities of all the compounds were determined for human beta(1)-ARs expressed in Chinese hamster ovary cells and the antagonist potency for rat atrial beta(1)-ARs was determined for 32 of these compounds for comparative purposes. The compounds, based upon a phenoxypropanolamine core structure with various meta-, ortho-, para- and amine-substituents, displayed binding affinities (pK(i)) for the human beta(1)-AR ranging from 5.49 to 9.35. Antagonist potencies (pA(2)) in the rat ranged from 5.52 to 9.56 and correlated with the human binding affinities (r(2)=0.86). Twenty-six compounds were used as the training set for comparative molecular field analysis (CoMFA) of antagonist binding affinity at the human beta(1)-AR and also of antagonist potency for rat atrial beta(1)-ARs. The CoMFA models were derived using both the CoMFA electrostatic and steric field parameters. The initial human beta(1)-AR model (n=26, q(2)=0.59, ONC=6, SE(CV)=0.70, r(2)=0.98, SE(non-CV)=0.16, F(6,19)=148) predicted the binding affinities of seven out of ten test compounds, not included in the training set, with residual pK(i) values less-than-or-equal0.50. The final human beta(1)-AR model (n=36, q(2)=0.66, ONC=5, SE(CV)=0.61, r(2)=0.95, SE(non CV)=0.24, F(5,30)=107), consisting of the training set plus the test set of compounds, may prove useful in the design of new phenoxypropanolamine type beta(1)-AR antagonists. The initial rat beta(1)-AR model (n=26, q(2)=0.42, ONC=6, SE(CV)=0.76, r(2)=0.94, SE(non-CV)=0.25, F(6,19)=47) predicted the affinities of five out of six test compounds with residual pA(2) values less-than-or-equal0.50. The final rat beta(1)-AR model (i.e. training set plus test set of compounds) (n=32, q(2)=0.38, ONC=5, SE(CV)=0.69, r(2)=0.93, SE(non-CV)=0.24, F(5,26)=67) in particular has a low q(2) value, indicating that, at least for the rat, the biologically active phenoxypropanolamine conformation may be quite different to the low energy extended conformation chosen for this CoMFA study. PMID- 11858846 TI - Conformational diversity of T-kinin in DMSO, water and HFA. AB - T-kinin (Ile-Ser-BK) is related to BK in its biological profile, but unlike BK, is more resistant to the action of ACE. A detailed NMR and molecular modeling study of T-kinin has been carried out in three diverse media: water (pH 4.0), DMSO-d(6) and HFA solution. In DMSO-d(6), T-kinin adopts a structure with two tandem beta-turns: the first at the mid segment tetrad Pro(4)-Pro(5)-Gly(6) Phe(7) (type I) and the C-terminal end Ser(8)-Pro(9)-Phe(10)-Arg(11) harbors the second turn (also type I). While the first beta-turn is lost in presence of water, the second persists. In HFA, NMR reveals a alpha-helix like structure spanning residues Arg(3) to Arg(11). Structures with cis peptide bonds (XX-Pro) have been observed for T-kinin in DMSO-d(6) but not in water and HFA. Differences in the structures of BK and T-kinin in water may explain their susceptibility/resistance to the action of ACE. PMID- 11858847 TI - COX-1/COX-2 inhibitors based on the methanone moiety. AB - This paper focuses on the synthesis and the in vitro testing of dual COX-1/COX-2 inhibitors. Starting from structures of non-steroidal anti-inflammatory drugs (NSAIDs) the diaryl methanone element was chosen as a lead. Modifications were carried out on this scaffold to obtain potent inhibitors of the COX enzymes. The N-(2-aroylphenyl)sulphonamides and -amides were studied in detail, and to consolidate the data evaluated the corresponding 3- and 4-regioisomers were also investigated. The potency and the enzyme selectivity were varied by structural modifications of the lead. PMID- 11858848 TI - New isoxazole derivatives designed as nicotinic acetylcholine receptor ligand candidates. AB - In this work we report the synthesis and evaluation of the analgesic properties of new isosteric heterocyclic derivatives, presenting the isoxazole nucleus, designed as nicotinic acetylcholine receptor ligand candidates, analogues to alkaloid epibatidine. Compound 2-(3-methyl-5-isoxazolyl)pyridine (3) presented the best analgesic profile of this series in hot plate test, which was partially prevented by pretreatment with nicotinic receptor antagonist mecamylamine. PMID- 11858849 TI - Synthesis and dopamine transporter binding of 2beta-isopropyl ester analogs of cocaine. AB - A series of 2beta-isopropyl ester analogs of cocaine (7-11) was synthesised and evaluated in an in vitro dopamine transporter (DAT) binding assays. Ecgonine HCl (5) was obtained from (-)-cocaine (1) by hydrolysis using 1 N HCl. Acid catalysed esterification of 5 using 2-propanol and HCl gas afforded 2beta-isopropyl ecgonine (6). Compounds 7-9 were obtained via esterification of the 3beta hydroxyl group of 6 using the appropriate acid chloride. Compound 10 was obtained via selective hydrolysis and re-esterification of 7 using 2-propanol and HCl gas. Compound 11 was obtained by reduction of 9 using H(2)/Pd-C. Compounds 7, 10 and 11 showed high binding affinity to the DAT (as indicated from the inhibition of the binding of [(3)H]WIN 35,428 (3)) with IC(50) values (mean +/- S.E.M.) 208.5 +/- 9.5, 47.43 +/- 1.79 and 11.25 +/- 3.37 nM, respectively). Compound 7 is comparatively as active as cocaine, 10 is ca. fivefold more active than cocaine and 11 is ca. 20-fold more active than cocaine and even twice more active than the radioligand 3. Compound 11, like its methyl ester analog (2' aminococaine), exhibited the highest affinity to the DAT. These results, along with previous results, emphasise the importance of a hydrogen-bond donor group at the 2' position of cocaine and its isopropyl ester analogs to enhance binding affinity to the DAT. PMID- 11858850 TI - Cytotoxic-antineoplastic activity of hydroquinone derivatives. AB - Several myrcenylhydroquinone derivatives have been evaluated for their cytotoxic activity against three neoplastic cell line cultures and compared with the activity previously observed on other neoplastic systems. Also a new series of this type of compounds has been prepared and the compounds synthesised have been evaluated by their GI(50) values. PMID- 11858851 TI - Strengthening immunization systems and introduction of hepatitis B vaccine in central and eastern Europe and the newly independent states. PMID- 11858852 TI - The utility of serologic testing for varicella in an adolescent population. PMID- 11858853 TI - What is a safe vaccine? PMID- 11858854 TI - Protection against feline immunodeficiency virus using replication defective proviral DNA vaccines with feline interleukin-12 and -18. AB - A molecular clone of the Glasgow-8 isolate of FIV (FIVGL8) was rendered replication defective by an in-frame deletion in either reverse transcriptase (deltaRT) or integrase (deltaIN) genes for use as DNA vaccines. To test the ability of these multi-gene vaccines to protect against two feline immunodeficiency virus (FIV) isolates of differing virulence, cats were immunized using either DNA vaccine alone or co-administered with interleukin-12 (IL-12) and/or interleukin-18 (IL-18) cytokine DNA. Animals were challenged sequentially with FIV-Petaluma (FIVPET) an FIV isolate of relatively low virulence and subsequently with the more virulent FIVGL8. A proportion of vaccinates (5/18 deltaIN and 2/12 deltaRT) were protected against primary challenge with FIV(PET). Five of the vaccinated-protected cats were re-challenged with FIV(PET); four (all deltaIN) remained free of viraemia whilst all naive controls became viraemic. Following subsequent challenge with the more virulent FIVGL8 these four vaccinated-protected animals all became viraemic but showed lower proviral loads than naive cats. This study suggests that while our current DNA vaccines may not produce sterilizing immunity against more virulent isolates of FIV, they may nevertheless significantly reduce the impact of infection. PMID- 11858855 TI - Monoclonal antibody protects mice against infection and disease when given either before or up to 24 h after airborne challenge with virulent Venezuelan equine encephalitis virus. AB - Airborne infection with Venezuelan equine encephalitis virus (VEEV) is a significant hazard for laboratory workers, who may not be immunised against VEEV infection as there is no vaccine currently available suitable for human use. We describe a potential alternative strategy that could protect workers exposed to VEEV or similar viruses. VEEV-specific murine monoclonal antibodies (MAB), given by intraperitoneal (i.p.) injection to mice as a single dose of 100 microg, have a half-life of 6-10 days in serum and spread by transudation to respiratory secretions. Administration of MAB (approximately 4 mg/kg) to mice 24h before challenge with approximately 100LD50 of virulent VEEV protected up to 100% animals. The same dose of MAB delivered up to 24h after challenge protected approximately 50%. Two MAB that were synergistic in vitro in plaque reduction neutralisation tests were not synergistic in vivo in protection assays. An examination of virus multiplication, in the blood and internal organs (brain, spleen, lung) of MAB-treated mice infected by the airborne route with VEEV, suggested that therapeutic activity depended both upon the prevention of virus infection of the brain, and the rapid clearance of virus from the periphery. Antiviral therapy with VEEV-specific human or "humanised" MAB, providing that they are administered early, may offer an alternative means of specific medical intervention for those with a known exposure to VEEV. PMID- 11858856 TI - A review of emergency foot-and-mouth disease (FMD) vaccines. AB - The primary objectives of this paper are to describe emergency foot-and-mouth disease (FMD) vaccines and review literature on emergency vaccine efficacy to protect animals against (1) clinical signs and (2) infection (local virus replication). The reviewed experiments suggest that in cattle, sheep and pigs, the vaccine could be effective in preventing disease within 4-5 days post vaccination. These studies also suggest that the risk of spreading infection decreases as the interval between vaccine and challenge increases and that vaccination could reduce the amount of virus excreted compared to non-vaccinated animals. We suggest areas of future research to improve our knowledge of emergency vaccines. PMID- 11858857 TI - Analysis of recombinant mouse zona pellucida protein 2 (ZP2) constructs for immunocontraception. AB - In this study we have examined the potential of recombinant mouse zona pellucida glycoprotein 2 (ZP2) as a target for immunocontraception. Immunogenicity studies and fertility trials were performed in outbred Swiss-Webster mice using four ZP2 constructs: Val(35)-Gly(200) (ZP2(V35-G200)), Val(35)-Leu(331) (ZP2(V35-L331)), Pro(325)-Ala(637) (ZP2(P325-A637)), and Val(35)-Ala(637) (ZP2(V35-A637)). A significant antibody response occurred to three of the four immunogens, however antibodies capable of recognizing native ZP occurred only after immunization with ZP2(V35-A637) and ZP2(P325-A637). Only immunization with ZP2(V35-A637) correlated with a reduction in fertility. Examination of the physiological basis for infertility revealed that: (1) passive transfer of ZP2 antiserum induced infertility in non-immune mice; (2) ovaries of infertile mice appeared histologically normal; (3) infertile mice produced normal numbers of eggs and (4) ZP of ovulated eggs from infertile mice demonstrated a significant reduction in the number of sperm bound compared to eggs from adjuvant controls. Infertility can be caused entirely by ZP2 antibodies without the incidence of significant ovarian pathology. This study also demonstrated that immunization with the bioactive (sperm binding) region of ZP2, recombinant ZP2(V35-G200), did not result in a significant immune response that recognized native ZP or inhibited fertility. Consequently we designed a ZP2-sperm antigen construct, replacing the C-terminal region of ZP2 with Sp17. This construct proved to be immunogenic and reduce fertility while directing the immune response to the Val(35)-Gly(200) region of ZP2. PMID- 11858858 TI - Protective immune responses elicited in mice by immunization with formulations of poly(lactide-co-glycolide) microparticles. AB - Parenteral administration of microparticle encapsulated DNA elicits immune responses to the encoded antigens. Experiments were performed to test whether the addition of certain lipophilic agents to such formulations enhanced the activity of a beta-galactosidase (beta-gal) DNA vaccine. Addition of either taurocholic acid (TA) or monomethoxy polyethylene-glycol-distearoylphosphatidylehanolamine (PEG-DSPE) increased the efficiency of DNA encapsulation. Immunization of mice with encapsulated DNA formulations containing either compound significantly increased the number of antibody positive responders over that achieved with non lipid containing particles. Moreover, responding animals demonstrated trends towards higher antibody titers and increased T cell responses. Tumor protection against the CT26.CL25 tumor cell line was demonstrated with lipid and non-lipid containing formulations. These results are the first demonstration of protection obtained by parenteral administration of PLG encapsulated DNA vaccines. PMID- 11858859 TI - Application of heat killed Mycobacterium bovis-BCG into the lung inhibits the development of allergen-induced Th2 responses. AB - We have previously reported that an infection of the lung with BCG-inhibited ovalbumin (OVA)-induced airway eosinophilia. In the current study, we investigated if the intranasal application of heat killed (HK)-BCG or purified protein derivative (PPD) from Mycobacterium tuberculosis had the same effect. For this purpose we treated mice intranasally with either live BCG, HK-BCG or PPD and analyzed if the mice developed airway eosinophilia after immunization and intranasal challenge with OVA. Our results clearly showed that an intranasal vaccination with live and HK-BCG but not PPD, given 4 or 8 weeks prior to allergen airway challenge, resulted in a strong suppression of airway eosinophilia. The inhibition of airway eosinophilia correlated with reduced levels of IL-5 production by T cells from the lymph node of the lungs and a strong reduction in Th2 cell numbers present in the airways of OVA-challenged mice. Furthermore, HK-BCG-induced suppression of airway eosinophilia was strongly reduced in IFN-gamma deficient mice. HK-BCG in contrast to live BCG may also be a promising candidate for a prospective asthma vaccine in humans since negative side effects due to mycobacterial infection can be ruled out. PMID- 11858860 TI - Detection of measles vaccine in the throat of a vaccinated child. AB - Measles vaccine is widely used, most often in association with mumps and rubella vaccines. We report here the case of a child presenting with fever 8 days after vaccination with a measles-mumps-rubella vaccine. Measles virus was isolated in a throat swab taken 4 days after fever onset. This virus was then further genetically characterised as a vaccine-type virus. Fever occurring subsequent to measles vaccination is related to the replication of the live attenuated vaccine virus. In the case presented here, the vaccine virus was isolated in the throat, showing that subcutaneous injection of an attenuated measles strain can result in respiratory excretion of this virus. PMID- 11858861 TI - Experimental non-transmissible marker vaccines for classical swine fever (CSF) by trans-complementation of E(rns) or E2 of CSFV. AB - Three mutants with deletions in the E2 gene of the infectious DNA copy of the classical swine fever virus (CSFV) strain-C were constructed: one missing the B/C domain of CSFV-E2 between amino acids (aa) 693 and 746, one missing the A domain between aa 800 and 864, and one missing the complete E2 between aa 689 and 1062. All three CSFV-E2 deletion mutants were unable to generate viable virus, indicating that each of the antigenic domains of E2 is essential for viability of CSFV. To rescue the CSFV-E2 deletion mutants SK6 cell lines constitutively expressing glycoprotein E2 of CSFV were generated. The rescued viruses infected and replicated in SK6 cells as demonstrated by expression of viral proteins, but this primary infection did not result in reproduction of infectious virus. Thus, these E2 complemented viruses are considered non-transmissible. In previous experiments, we showed that simultaneous injection of E(rns) complemented virus (Flc23) via intradermal (ID), intramuscular (IM) or intranasal (IN) routes conferred protection to pigs against a lethal challenge with CSFV [J. Virol. 74 (2000) 2973]. Here, we evaluate different routes of application (ID, IM or IN) with E(rns) complemented virus Flc23 in order to find the best route for complemented CSFVs. Intradermal injection with Flc23 protected pigs against a lethal CSFV challenge, whereas intramuscular injection induced partial protection, and intranasal injection did not mediate a protective immune response in pigs at all. We used the intradermal route of vaccination to test the E2 complemented viruses. Vaccination of pigs via the intradermal route with the E2 complemented CSFVs also resulted in the induction of antibodies and in (partial) protection against CSFV challenge. Pigs vaccinated with E2 complemented virus Flc4 (deletion B/C domain) survived a lethal CSFV challenge, whereas partial protection was induced in pigs vaccinated with Flc47 (deletion E2) or Flc48 (deletion A domain) E2 complemented viruses. Serological data demonstrate that these E2 complemented mutant viruses are, in combination with well known diagnostic tests based on E2, potential marker vaccines for CSF. PMID- 11858862 TI - Immunogenicity of hepatitis B vaccine in preterm and full term infants vaccinated within the first week of life. AB - The immunogenicity of a Hepatitis B vaccine was evaluated in 110 neonates (57 full term and 53 preterm) born to Hepatitis B surface antigen (HBsAg) negative mothers. Three 10 microg doses of recombinant Hepatitis B vaccine were administered: the first dose within the first week of life; the second between 1 and 2 months; and the third at 5-7 months of age. Anti-HBs antibody titres were measured 3 months after the third dose. The seroconversion rate in preterm infants (77%; 95% CI=64.7-87.1) was significantly lower than in full term infants (98%; 95% CI=91.6-99.9) while the mean anti-HBs titres among those infants that did seroconvert was lower in preterm (186.6 mIU ml(-1)) than in full term infants (537.5 mIU ml(-1)). More full term than preterm infants showed titres greater than 100 mIU ml(-1) (71.9 and 41.5%, respectively). We conclude that the administration of a recombinant Hepatitis B vaccine shortly after birth is less immunogenic in preterm infants weighing <1800 g at birth than in full term infants. Currently accepted recommendations for post exposure perinatal prophylaxis may be inadequate to protect preterm infants. PMID- 11858863 TI - Protective efficacy of a plasmid DNA encoding Japanese encephalitis virus envelope protein fused to tissue plasminogen activator signal sequences: studies in a murine intracerebral virus challenge model. AB - We report the construction of chimeric DNA vaccine vectors in which secretory signal sequence derived from tissue plasminogen activator (TPA) was fused to the full length (pCMVTE) or 398 amino terminal amino acids (pCMVTdeltaE) of Japanese encephalitis virus (JEV) envelope (E) protein. Transfection studies indicate that E protein expressed from pCMVTdeltaE-transfected cells but not pCMVTE-transfected cells is secreted into the culture medium. Analysis of the potency of various DNA vaccine constructs in a murine intracerebral (i.c.) JEV challenge model indicates that pCMVTdeltaE confers the highest level (71%) of protection. Immunization with pCMVTdeltaE induces a mixed Th1 and Th2 T helper cell response while immunization with plasmids encoding nonsecretory forms of E protein induces a Th1 T helper response. Only low levels (<1:20) of virus neutralizing antibody titres were observed in DNA vaccinated mice which did not increase further after i.c. JEV challenge. Thus, immunization with a plasmid encoding secretory E protein results in an altered cytokine response and better protection against i.c. JEV challenge than that conferred by immunization with plasmids encoding nonsecretory forms of E protein. We also demonstrate that unlike peripheral JEV challenge, i.c. JEV challenge does not result in an increase in anamnestic antibody response suggesting that other components of immune system such as cytotoxic T cells and T helper cells contribute to protection against i.c. JEV challenge of DNA vaccinated mice. PMID- 11858864 TI - Immunization of T-cell deficient mice against polyomavirus infection using viral pseudocapsids or temperature sensitive mutants. AB - A murine experimental model system aimed at developing potential vaccines to papovavirus infection in immunosuppressed individuals was explored. A VP1 pseudocapsid based on the major capsid protein of the murine polyomavirus A2 strain and a mutant, M17-pseudocapsid as well as four temperature sensitive (ts) mutants were used as immunogens. T-cells deficient CD4-/-8-/- mice were immunized four times with each immunogen and then together with non-immunized control mice challenged with polyomavirus. In contrast to all control mice, only half of the immunized mice exhibited presence of polyoma DNA when assayed by PCR. The results indicate that pseudocapsids and ts-mutant immunization may potentially protect mice with an impaired T-cell function from polyomavirus infection. PMID- 11858865 TI - Intranasal inoculation of a recombinant influenza virus containing exogenous nucleotides in the NS segment induces mucosal immune response against the exogenous gene product in mice. AB - To establish a mucosal vaccine system, we evaluated the immunogenicity of a recombinant influenza virus, designated NS2Acat, in which the chloramphenicol acetyltransferase (CAT) reporter gene is fused in frame to the NS1 gene in the NS gene segment. The NS2Acat replicated slightly within the lungs in BALB/c mice after intranasal administration, accompanying expression of the CAT and the viral HA mRNA. NS2Acat induced delayed-type hypersensitivity (DTH) and sensitized the CAT-specific T lymphocytes in the regional lymph nodes, which proliferated and synthesized several cytokines in vitro upon antigenic challenge. These results suggest that NS2Acat is capable of eliciting a respiratory immune response against the fused gene product. PMID- 11858866 TI - Enhanced immune protection by a liposome-encapsulated recombinant respiratory syncytial virus (RSV) vaccine using immunogenic lipids from Deinococcus radiodurans. AB - The radiation-resistant bacterium, Deinococcus radiodurans contains a variety of phospho-, glyco- and phosphoglycolipids, the structures of which appear to be largely unique in nature. We show here that such lipids are immunogenic when administered as liposomes intranasally in mice, as evidenced by the induction of serum antibodies which recognize D. radiodurans lipids but not other lipids by thin layer chromatographic immunostaining. By modifying a liposomal vaccine against respiratory syncytial virus (RSV) we find that vaccine efficacy is markedly enhanced by the inclusion of lipids isolated from D. radiodurans. Using dioleoylphosphatidylcholine (DOPC) or D. radiodurans lipids, liposomes were prepared which encapsulated a soluble fragment of the RSV G protein (G(128-188)) fused with a portion of the bacterial thioredoxin (Trx) protein. Mice immunized intranasally with D. radiodurans liposomes showed markedly greater protection against RSV challenge over mice immunized with DOPC liposomes. Enhanced vaccine efficacy was achieved using liposomes prepared from either whole D. radiodurans lipids or from a single isolated phosphoglycolipid previously identified as alpha galactosylphosphatidylglyceroylalkylamine (lipid 7). Mice immunized and protected against RSV challenge were free of pulmonary eosinophilic infiltration, an undesirable consequence of many RSV vaccines. The results provide further support for liposome-based vaccines for RSV and underline the importance of lipid composition in liposome formulations. PMID- 11858868 TI - Adverse event reports following vaccination for Lyme disease: December 1998-July 2000. AB - CONTEXT: The vaccine adverse event reporting system (VAERS) monitors vaccine safety post-licensure. Although events reported to VAERS are not necessarily causally associated with vaccination, VAERS reports can be used to identify possible safety concerns that occur at too low a rate to have been identified prior to licensure. OBJECTIVE: To evaluate adverse events following Lyme disease vaccination reported to VAERS during the first 19 months of the vaccine's licensure. DESIGN, SETTING, AND PARTICIPANTS: Analysis of all VAERS reports of adverse events following vaccination for Lyme disease in the US from 28 December 1998 to 31 July 2000. MAIN OUTCOME MEASURE: We evaluated reported adverse events for unexpected patterns in age, gender, time to onset, dose number, and clinical characteristics and compared them to adverse events observed in clinical trials of this vaccine. RESULTS: Over 1,400,000 doses were distributed and 905 adverse events were reported to VAERS, 440 in men and 404 in women, with ages ranging from 10 to 82 years. The majority (56%) of adverse events occurred after administration of the first dose. The most frequently reported adverse events were arthralgia (250), myalgia (195), and pain (157). There were 59 reports coded as arthritis, 34 as arthrosis, 9 as rheumatoid arthritis, and 12 as facial paralysis. Sixty-six (7.4%) events were classified as serious, involving life threatening illness, hospitalization, prolongation of hospitalization, persistent or significant disability/incapacity, or death. Twenty-two hypersensitivity reactions were reported. CONCLUSION: Based on reporting to VAERS, we did not detect unexpected or unusual patterns of reported adverse events following Lyme disease vaccine administration, other than hypersensitivity reactions, compared with adverse events observed in clinical trials. PMID- 11858867 TI - Immunogenicity of a recombinant varicella-zoster virus gE-IE63 fusion protein, a putative vaccine candidate against primary infection and zoster reactivation. AB - The varicella-zoster virus (VZV) envelope glycoprotein E (gE) and immediate early protein 63 (IE63) are well known targets for specific humoral and cell-mediated immune responses during VZV infection and latency, respectively. The present study evaluated the immunogenicity of an engineered chimeric recombinant gE-IE63 (recgE-IE63) protein secreted from CHO cells, wherein a soluble form of gE, deleted of its anchor and cytoplasmic domains was fused to IE63. Guinea pig vaccinations with adjuvanted recgE-IE63 elicited a strong and specific humoral immune response directed to each counterpart. Sera from recgE-IE63-immunized animals neutralized cell-free VZV. This neutralizing capacity was dependent only on the recgE moiety as serum depletions on recgE-immobilized sepharose totally abolished VZV neutralization. The cell-mediated immune response induced by recgE IE63 was evaluated in lymphoproliferation assays. An antigen-specific proliferative response was demonstrated after lymphocyte stimulation with recIE63 but not with recgE. We conclude that recombinant chimeric recgE-IE63 induced both humoral and cell-mediated immune responses and thus could constitute a putative subunit vaccine candidate against VZV primary infection and zoster reactivation. PMID- 11858871 TI - Evidence based vaccinology. AB - Evidence based vaccinology (EBV) is the identification and use of the best evidence in making and implementing decisions during all of the stages of the life of a vaccine, including pre-licensure vaccine development and post-licensure manufacture and research, and utilization of the vaccine for disease control. Vaccines, unlike most pharmaceuticals, are in a continuous process of development both before and after licensure. Changes in biologics manufacturing technology and changes that vaccines induce in population and disease biology lead to periodic review of regimens (and sometimes dosage) based on changing immunologic data or public perceptions relevant to vaccine safety and effectiveness. EBV includes the use of evidence based medicine (EBM) both in clinical trials and in national disease containment programs. The rationale for EBV is that the highest evidentiary standards are required to maintain a rigorous scientific basis of vaccine quality control in manufacture and to ensure valid determination of vaccine efficacy, field effectiveness and safety profiles (including post licensure safety monitoring), cost-benefit analyses, and risk:benefit ratios. EBV is increasingly based on statistically validated, clearly defined laboratory, manufacturing, clinical and epidemiological research methods and procedures, codified as good laboratory practices (GLP), good manufacturing practices (GMP), good clinical research practices (GCRP) and in clinical and public health practice (good vaccination practices, GVP). Implementation demands many data driven decisions made by a spectrum of specialists pre- and post-licensure, and is essential to maintaining public confidence in vaccines. PMID- 11858870 TI - Immunogenicity of chi4127 phoP- Salmonella enterica serovar Typhimurium in dogs. AB - Salmonellae are commonly isolated from dogs. The number of dogs infected with Salmonella spp. is surprisingly high and greater than the incidence of clinical disease would suggest. Salmonellosis is common in greyhound kennels. Morbidity can approach 100% in puppies and the mortality ranges to nearly 40%. To date, there has been little effort to evaluate the feasibility of a vaccine for control of this disease in dogs. In the studies described here, an attenuated strain of Salmonella enterica serovar Typhimurium (Se Typhimurium), chi4127, was capable of establishing a limited infection in dogs. The chi4127-attenuated salmonellae efficiently stimulated protective immune responses in serotype homologous, direct, oral challenge experiments. Morbidity in the wild-type-challenged dogs was 8.3% in immunized dogs but 100% in the non-vaccinated controls. In (9/12) control dogs, the disease involved both gastrointestinal and respiratory tracts with high fever (>40.2 degrees C) that persisted through 5 days after challenge. Serum IgG response against S. typhimurium lipopolysaccharide (LPS) significantly increased (P<0.01) in vaccinated dogs and in non-vaccinated dogs after challenge. The non-vaccinated dogs had 3 to 4 logs higher numbers of Se Typhimurium in splenic and hepatic tissue than did the vaccinated dogs. This particular attenuated strain has potential for use as a vaccine for canine salmonellosis. PMID- 11858869 TI - Alphavirus replicon particles expressing the two major envelope proteins of equine arteritis virus induce high level protection against challenge with virulent virus in vaccinated horses. AB - Replicon particles derived from a vaccine strain of Venezuelan equine encephalitis (VEE) virus were used as vectors for expression in vivo of the major envelope proteins (G(L) and M) of equine arteritis virus (EAV), both individually and in heterodimer form (G(L)/M). The immunogenicity of the different replicons was evaluated in horses, as was their ability to protectively immunize horses against intranasal and intrauterine challenge with a virulent strain of EAV (EAV KY84). Horses immunized with replicons that express both the G(L) and M proteins in heterodimer form developed neutralizing antibodies to EAV, shed little or no virus, and developed only mild or inapparent signs of equine viral arteritis (EVA) after challenge with EAV KY84. In contrast, unvaccinated horses and those immunized with replicons expressing individual EAV envelope proteins (M or G(L)) shed virus for 6-10 days in their nasal secretions and developed severe signs of EVA after challenge. These data confirm that replicons that co-express the G(L) and M envelope proteins effectively, induce EAV neutralizing antibodies and protective immunity in horses. PMID- 11858872 TI - Early protection against homologous challenge after a single dose of replication defective human adenovirus type 5 expressing capsid proteins of foot-and-mouth disease virus (FMDV) strain A24. AB - Previously we demonstrated that two doses of a replication-defective human adenovirus serotype 5 (Ad5) carrying the capsid (P1) and 3C protease coding regions of a laboratory strain of FMDV (A12) completely protected five of six swine challenged with homologous virus. The objective of the current study was to evaluate the efficacy of one dose of an Ad5-vectored vaccine expressing the P1 coding region of an FMDV field strain. A replication-defective Ad5 containing the P1 coding region of FMDV A24 and the 3C coding region of A12 (Ad5A24) was constructed and evaluated for its ability to induce neutralizing antibodies and protect swine against homologous challenge after a single vaccination. Animals were challenged 7, 14 or 42 days after vaccination. Control groups included animals inoculated with commercial vaccine or phosphate-buffered saline. All vaccinated swine were completely protected against homologous challenge at 7, 14 or 42 days after vaccination. Based on these results, we conclude that a single inoculation of Ad5-vectored vaccines could be used as a tool to control FMD in outbreak situations. PMID- 11858873 TI - Genetic and immunological comparison of anti-botulinum type A antibodies from immune and non-immune human phage libraries. AB - Understanding the antibody response in botulinum intoxication is important for vaccine design and passive prophylaxis. To investigate this activity, we have studied the immune response to BoNT/A (botulinum neurotoxin serotype A) binding domain (HC) at the molecular level using phage display. The scFv antibodies were isolated from V-gene repertoires prepared from (a) human volunteer immunized with pentavalent botulinum toxoid and (b) non-immune human peripheral blood lymphocytes and spleenocytes. A large panel of serotype specific phage expressing botulinum binding scFv could be selected from both libraries. Epitope mapping of immune scFv binders towards BoNT/A HC revealed surprisingly a limited number of scFv recognizing conformational epitopes that corresponded to two distinct groups, clusters I and II. Only scFv from cluster I exhibited neutralizing activity in the mouse hemidiaphragm assay. Anti- BoNT/A HC clones derived from a non-immune library could be conveniently grouped into clusters III-XI and appeared to share no overlapping epitopes with cluster I or II. In addition they showed no neutralization of toxin at biologically significant concentrations. We therefore suggest that a vaccine based on the pentavalent botulinum toxoid directs the humoral immune response to a limited number of immunodominant epitopes exposed on the binding domain HC. PMID- 11858874 TI - Clinical trial of the intradermal administration of hepatitis B vaccine produced at the Department of Medical Research, Myanmar. AB - A total of 280 apparently healthy volunteers were screened for hepatitis B (HB) markers out of which 49 subjects (17.5%) were positive for HB surface antigen (HBsAg) and 82 (29.3%) were positive for antibody to HBsAg (anti-HBs). Three doses of DMR-HB vaccine, 0.15 ml per dose were administered to 95 subjects, who were serologically negative for both HB markers. The vaccination was given by the intradermal route on the flexor surface of the left forearm, at 1 month intervals according to the 0, 1 and 2 months schedule. The subjects were carefully monitored to record any adverse reaction of the vaccine. Blood specimen was collected from each subject, 1 month after the second and third vaccinations, to determine the anti-HBs antibody response to the vaccine. The study results showed that local pain was the only side effect noted and protective antibodies (anti HBs) were detected in 69 (72.6%) of the vaccinees after the second dose of the vaccine and 89 (93.6%) after the third dose of the vaccine. Thus the intradermal route, which would require approximately one-seventh of the standard dose, would be suitable for use in certain groups such as high risk adults, when the cost of the vaccine is the inhibiting factor for routine or mass vaccination. PMID- 11858875 TI - Nasal mucosal immunogenicity for the horse of a SeM peptide of Streptococcus equi genetically coupled to cholera toxin. AB - The intranasal immunogenicity of cholera toxin (CT) genetically coupled to peptide sequence aa236-334 (F3) of the SeM protein of Streptococcus equi was studied in five young adult Welsh ponies. All ponies made rapid CTB- and SeMF3 specific serum antibody responses following the first immunization. Specific nasal IgA responses were detected in two ponies 14 days after the first immunization, in another two 14 days after a second immunization on day 14, and in all ponies 28 days after a third immunization on day 42. SeMF3-specific antibody responses in sera and nasal washes were dominated by IgGb and IgA, respectively, and remained elevated for at least 140 days. Strong serum IgGa and IgG(T) responses were also observed. These antibody responses were qualitatively similar to those induced during recovery from equine strangles. Antibody responses in mucosal secretions were boosted in some ponies by immunizations subsequent to the first immunization, but antibodies in serum were never boosted. In vitro survival of S. equi was significantly reduced by SeMF3-specific antibodies in sera obtained 14 days after the second immunization but survival increased in sera collected following subsequent immunizations, possibly due to absence of synthesis of high affinity antibodies. Finally, the susceptibility of all immunized ponies to commingling challenge by S. equi indicated either that SeMF3 lacks protective epitopes or that the antibodies induced by the chimera were not at effective levels. PMID- 11858876 TI - Immunogenicity of the Brucella melitensis recombinant ribosome recycling factor homologous protein and its cDNA. AB - A study was conducted to evaluate the immunogenicity of the Brucella melitensis ribosome recycling factor (RRF)-homologous protein (CP24). The CP24 gene was cloned, expressed in Escherichia coli and purified. The resulting purified recombinant protein (rCP24) produced delayed-type hypersensitivity (DTH) reactions in B. melitensis-infected mice but not in naive controls. Thus, we decided to characterise the immune responses generated with DNA vaccination (pcDNACP24) or immunisation with the rCP24 in adjuvant. Animals injected with pcDNACP24 exhibited a dominance of IgG2a to IgG1 while mice injected with rCP24 developed a higher response of IgG1 than IgG2a. Both immunisation protocols were capable of eliciting CP24-specific gamma interferon (IFN-gamma) producing cells. Spleen cells from pcDNACP24-immunised mice did not produce interleukin (IL)-4, IL 10 or up-regulation of IL-2 mRNA. Cells from rCP24-immunised mice produced IL-10, up-regulated IL-2 mRNA but did not produce IL-4. Neither immunisation with purified CP24 nor injection of pcDNACP24 protected mice against challenge with live smooth B. melitensis. However, the potential of CP24 for a Brucella diagnostic test based on an in vitro antigen (Ag)-specific IFN-gamma production or DTH test would be worth testing. PMID- 11858877 TI - Supplementation of conventional trivalent influenza vaccine with purified viral N1 and N2 neuraminidases induces a balanced immune response without antigenic competition. AB - Influenza viruses neuraminidase (NA) were chromatographically extracted from influenza viruses A/Nanchang/933/95 H3(NC)N2(NC) [R] and A/Johannesburg/82/96 H1(JH)N1(JH) [R] and used to supplement conventional inactivated trivalent influenza vaccine. Immunization of mice with this preparation resulted in high titers of antibodies to both hemagglutinins (HA) and neuraminidases (NA); there were no significant differences in the anti-HA antibody titers between the conventional and the supplemented vaccine preparation. Likewise, there were no significant differences in anti-NA antibody titers between the supplemented vaccine and titers from mice immunized with a neuraminidase vaccine containing a mixture of N1-NA and N2-NA. There was no evidence of a diminution of the immune response to the HA components of the vaccine despite the presence of antigenically equivalent amounts of both N1-NA and N2-NAs. Homotypic and distantly related heterotypic infections for both H1, N1 and H3N2 subtypes were suppressed and greater reduction in pulmonary virus titers (PVT) were observed in the trivalent vaccine supplemented with purified neuraminidase from each subtype, N1 and N2. Effects on the influenza B viral components were not studied. Previous studies on supplementation of conventional influenza vaccine focused only on monovalent H3N2 vaccine preparations; this study demonstrates in a mouse model system that supplementation of trivalent influenza vaccine with both influenza A subtype neuraminidases, N1 and N2 is highly immunogenic for HA and NA of each subtype and efficacious in protecting against influenza from homotypic and heterotypic infectious challenges of either subtype. PMID- 11858878 TI - Absence of antigenic competition in Aotus monkeys immunized with Plasmodium falciparum DNA vaccines delivered as a mixture. AB - Aotus lemurinus lemurinus monkeys were immunized four times with one of three DNA plasmids expressing important Plasmodium falciparum blood stage vaccine candidate proteins or with a mixture containing all three vaccines. The three vaccines encoded sequences from apical merozoite antigen-1 (AMA-1), erythrocyte binding protein-175 (EBA-175) and merozoite surface protein-1 (MSP-1). Antigen-specific enzyme-linked immunosorbant assays (ELISAs) showed no significant differences in antibody titer induced to the three antigens by a single vaccine compared with the titer induced to that same antigen by the trivalent preparation. Results of immunofluorescent antibody assays against erythrocytes infected with asexual blood stage P. falciparum indicated that each of the three monovalent vaccines induced significant antibody responses to whole parasites. The trivalent vaccine mixture induced, after four immunizations, an antibody titer to whole parasites that was 3--12-fold higher than those induced by any of the single vaccines. The fourth immunization with the trivalent vaccine increased the mean antibody in IFAT by more than five-fold. PMID- 11858879 TI - Oral immunization of mice with ricin toxoid vaccine encapsulated in polymeric microspheres against aerosol challenge. AB - Mucosal (oral) immunization of mice with carrier-delivered ricin toxoid (RT) vaccine was accomplished by one long (7 weeks) or two short (4 weeks) immunization schedules. For the long and short immunization schedule two lots of vaccine were administered prepared with the same procedure but at different occasions. The long schedule consisted of a total of seven doses of 50 microg of vaccine in microencapsulated (lot #108) or aqueous form administered on days 1, 2, 3, 28, 29, 30 and 49. With the short schedule a total of seven or six doses of 25 microg (lot #111) were administered on days 1, 2, 3, 14, 15, 16 and 30, or on 1, 2, 14, 15, 30, 31 and 32, respectively. Mice immunized orally with the long schedule, 50 microg of RT vaccine incorporated into poly-DL-lactide-co-glycolyde (DL-PLG) microspheres (MS) produced serum IgG, IgG2a and IgA ELISA antibodies. All mice immunized with RT in DL-PLG MS (RT-MS) were protected against a lethal ricin aerosol challenge. In contrast, with the same schedule and with the same dose, the aqueous vaccine (RT) failed to stimulate IgG, IgG2a and IgA antibodies, and these mice were not protected against an aerosol ricin toxin challenge. With the shorter immunization scheme, seven doses of 25 microg RT-MS stimulated a significant, though reduced, protection with the microencapsulated, but not with the aqueous vaccine. When the first and second 3-day cycles of the short immunization schedule was reduced to two doses, and the 3-day cycle was administered at the end of the schedule, neither RT-MS nor RT stimulated protection against the challenge. These results indicated that successful oral immunization with RT-MS depended on both the dose and the schedule, consisting of three consecutive days of administration in two cycles, 4 weeks apart. Altering this schedule and the dose, resulted in a reduced protection or no protection at all. Furthermore, under the conditions of this study, the advantage of the microencapsulated RT vaccine over the aqueous vaccine for effective oral immunization was well demonstrated. PMID- 11858881 TI - Pinopode expression during human implantation. AB - Implantation failure is considered the major impediment to successful in vitro fertilization pregnancy rate. Despite recent advances in reproductive medicine, the role played by uterine receptivity in contributing to human infertility remains unclear. Several clinical studies suggest the importance of blastocyst implantation in a narrow window of uterine receptivity, between 6 and 8 days after ovulation. This brief and precise period, called "implantation or nidation window", lasts less than 48 h and coincides with the formation of large and smooth projections, called "pinopodes", on the apical membranes of the endometrial epithelial cells. Pinopode expression appears to advance or retard depending on the hormonal treatments and an individual's particular response. This displacement can be followed through endometrial biopsy and scanning electron microscopy (SEM). Therefore, pinopode expression could be considered a useful biological marker to assess endometrial receptivity and to locate the implantation window. PMID- 11858882 TI - Vulvar vestibulitis syndrome: an overview of non-surgical treatment. AB - Vulvar vestibulitis syndrome, which represents one of the major cause of dyspareunia, is a puzzling clinical entity. Although many treatment options have been employed, a rationale therapeutic strategy is still not stated. The present article reviews the most popular medical approaches of such entity (biofeedback, tricyclic antidepressants, interferon psychologic-behavioural therapy, diet modification), as well as those to avoid. Tricyclic antidepressants and biofeedback of the pelvic floor muscles represents the first line effective therapy. Moreover, psychological counselling must support any treatment options. PMID- 11858883 TI - Mifepristone: bioavailability, pharmacokinetics and use-effectiveness. AB - The potentiality of mifepristone as an abortifacient and contraceptive drug along with its pharmacokinetic parameters is reviewed. Mifepristone or RU486 acts as antagonist to progestational and glucocorticoid functions. It is an orally active compound with nearly 70% absorption rate but its bioavailability is reduced to around 40% because of the first-pass effect. Peak plasma concentrations of 1.9 +/ 0.8, 3.8 +/- 0.9 and 5.3 +/- 1.3 micromol/l are reached within 1-2 h after oral administration of 50, 200 and 600 mg mifepristone in women, respectively, and are maintained at relatively high level up to 48 or 72 h depending on the ingested dose. The plasma kinetics of mifepristone followed two-compartment open model with a mean alpha-half-life of 1.4h, volume of distribution 1.47 l/kg and beta half-life of 20-30 h in most of the subjects studied. Clearance from the body was mainly through feces (83%). Biologically active mono-demethylated, di demethylated and hydroxylated metabolites were found in plasma soon after oral administration of mifepristone. RU486 and its mono-demethylated metabolite bind to progesterone receptors with high affinity. Mifepristone-bound receptor dimers suppress transcription activation and thus, bring about anti-progestational activity that makes mifepristone a potential abortifacient and contraceptive agent. Clinical trials for termination of early pregnancy with 50-600 mg mifepristone plus a prostaglandin analogue achieved a success rate of 82-97%. However, abdominal pain, cramping, nausea, vomiting, bleeding and delay in onset of the next menstrual cycle were the side effects. Administration of 25 mg mifepristone twice 12h apart, as a post-coital contraceptive showed 100% contraceptive efficacy. A low dose of mifepristone which does not inhibit ovulation reduced fertility significantly by affecting endometrial milieu. These findings suggest that reduced dose(s) of mifepristone, 200 mg or less, may be used as a post-coital contraceptive and in combination with vaginal misoprostol for termination of early pregnancy with high efficacy and minimal or no side effects. PMID- 11858884 TI - Prediction of vaginal delivery following caesarean section for failure to progress based on the initial aberrant labour pattern. AB - OBJECTIVE: To determine whether the pattern of cervical dilatation prior to caesarean section for 'failure to progress' can predict the outcome of subsequent labour. STUDY DESIGN: Single hospital case note review of 171 women delivered by caesarean section for failure to progress and subsequently delivering at the same hospital. Cervicograms were categorized into one of the four patterns by an assessor blinded to the subsequent outcome. Statistical analysis was done by analysis of variance. RESULTS: The incidence of vaginal delivery did not significantly differ between the groups. CONCLUSION: Categorisation of failure to progress by partographic abnormality does not predict subsequent successful vaginal delivery. PMID- 11858885 TI - Increased nuchal translucency and congenital heart defects in euploid fetuses. The Szeged experience. AB - OBJECTIVE: To determine the utility of the first-trimester fetal nuchal translucency (NT) thickness in the prediction of fetal cardiac malformations. DESIGN: Retrospective study. SETTING: Department of Obstetrics and Gynecology and Medical Genetics, University of Szeged. METHODS: The pre- and postnatal course and outcome, and the relationship between the first-trimester fetal NT thickness and fetal congenital heart defects (CHDs) in 4309 pregnancies ended up with birth or therapeutic abortion between January 1998 and June 2000 were registered. Prenatal care included first- and second-trimester fetal sonography at weeks 10 13 and 18-20, respectively. RESULTS: 4251 births and 58 first- and second trimester therapeutic abortions due to lethal congenital malformations or chromosomal abnormalities were recorded. Altogether 209 (4.9%) congenital malformations were detected, 39 (18.7%) of which were heart defects with normal karyotype. At birth, 151 congenital malformations were diagnosed, 34 of them were known prenatally. The prevalence of CHDs was 9 per 1000 pregnancies. The measurement of fetal NT thickness was available in 35 of the 39 fetuses with heart defects: it was > or = 3 mm in 18 (51.4%) and <3 mm in 17 (48.6%). A sensitivity of 51.4% was found at a cutoff of 3mm. CONCLUSIONS: An increased NT thickness in chromosomally normal fetuses was found to be highly associated with CHDs and identified in more than half of the affected cases. Furthermore, an increased NT of > or = 3 mm can be regarded a selection criterion for early second-trimester targeted fetal echocardiography and for increased fetal and neonatal surveillance. PMID- 11858886 TI - Pregnancy in women with systemic lupus erythematosus. AB - BACKGROUND: Systemic lupus erythematosus (SLE) is an autoimmune disorder which may be affected by hormonal changes, such as those of pregnancy. Women with SLE have increased adverse pregnancy outcomes. STUDY DESIGN: A retrospective analysis of the gynecologic and immunologic case history of 140 women with SLE and the outcome of 263 pregnancies in 99 women with SLE. RESULTS: In patients diagnosed with SLE, the proportion of pregnancies ending with live birth at term decreased to one-third compared with three quarters in those without a diagnosis of SLE and the incidence of pre-term deliveries and spontaneous abortions increased by 6.8 and 4.7 times, respectively. When SLE was associated with secondary antiphospholipid (APL) syndrome, and lupus anticoagulant (LA) or beta2 glycoprotein antibodies were present, a further increase in the incidence of pregnancy loss was observed. Pregnancy did not cause a flare-up of SLE in all cases, the disease remained stable in about 30% of the patients. Lupus was mild in the majority of the women who carried out their pregnancy to term. We also observed mothers with active SLE who successfully carried out pregnancies to term. CONCLUSION: These findings accord with previous literature and should inform rheumatologists, obstetricians and neonatologists who guide patients in their reproductive decisions. PMID- 11858887 TI - Cyclic changes in the umbilical arterial flow in mono-chorionic, di-amniotic twin pregnancy. AB - OBJECTIVE: To determine the effect of arterio-arterial anastomosis on the umbilical arterial flow in mono-chorionic, di-amniotic twin pregnancy. SUBJECTS: Thirty-three women with mono-chorionic, di-amniotic twin pregnancy. METHODS: Pulsed Doppler ultrasonographic examination was performed every 4 weeks to obtain umbilical arterial flow velocity waveforms in both two fetuses. RESULTS: In five of the women, cyclic flow changes in the umbilical arterial flow were detected in the smaller fetus. All of these twins had arterio-arterial anastomosis on the surface of the placenta. CONCLUSION: The fetuses in mono-chorionic and di amniotic twin gestation who showed cyclic changes in umbilical arterial flow had arterio-arterial anastomosis. But, this characteristic flow was not observed every time. PMID- 11858888 TI - Vasoconstrictive activity of oxytocin in meconium impregnated human placentas. AB - OBJECTIVE: The aim of our study is to determine whether oxytocin acts differently on the fetal-placental vascular bed of normal and meconium impregnated placentas. STUDY DESIGN: Isolated placental cotyledons (n=10) were dually perfused with fetal perfusion pressure used as an index of vascular resistance. As perfusion medium we used lactated Ringer salt solution, containing polyvinylpyrolidone (25 g/l), bovine serum albumin (0.1 mg/ml), glucose (1.0 g/l), heparin (20 IU/ml) and gentamycin (48 microg/ml). The pH of the medium was adjusted to 7.4 with bicarbonate. The maternal site was gassed with 95% O(2):5% CO(2) and in the fetal site with 95% N(2):5% CO(2) at 37 degree C. Perfusion rates were 4-6 and 10-12 ml/min in the fetal and maternal circulation, respectively. TNF-alpha and IL beta1 levels in the fetal-placental perfusate were evaluated using specific commercial ELISA kits. RESULTS: No significant changes in the amount of TNF-alpha release were observed after injection of oxytocin into the fetal circulation (31+3pg/ml; P=0.5). No IL-beta1 activity was observed in the fetal perfusate of normal and meconium impregnated placentas during the experiments. No significant difference was seen in basal perfusion pressure in normal and meconium impregnated placentas, however, a bolus injection of oxytocin (10U/ml) resulted in a significant increase in perfusion pressure in meconium impregnated placentas from basal pressure of s45+5 to 88+4 mm Hg after injection of oxytocin, (P=0.004, ANOVA). CONCLUSION: Vasoconstrictive effect of oxytocin was observed only in meconium impregnated placentas and no vascular effect of oxytocin was documented in normal placentas. The clinical implication of our findings is that one should use oxytocin for stimulation of labor with caution in the presence of meconium stained amniotic fluid. PMID- 11858889 TI - Quantitative foetal fibronectin as a predictor of successful induction of labour in post-date pregnancies. AB - OBJECTIVE: To study the relationship between induced labour, quantitative levels of fibronectin and the Bishop score. STUDY DESIGN: Vaginal fibronectin/Bishop score were estimated in 33 nulliparous women undergoing induction of labour for post-dates at the Department of Obstetrics and Gynaecology, Guy's and St. Thomas' Hospital London. RESULTS: There was no significant relationship between either the fibronectin level or Bishop score and the duration of the latent phase (R(2)=0.001; P=0.86 and R(2)=0.12; P=0.08, respectively). There was no relationship between the total prostaglandin dose and fibronectin level (R(2)=0.03; P=0.39) nor any significant correlation between either the Bishop score or fibronectin level and the induction to delivery time (R(2)=0.13; P=0.11 and R(2)=0.0006; P=0.97, respectively). Significant relationships were observed inversely between the total prostin dose and Bishop score (R(2)=0.33; P=0.002), between the total prostin dose and latent phase (R(2)=0.54; P=0.000009) and between Bishop score and the fibronectin levels (R(2)=0.19; P<0.01). CONCLUSIONS: Quantitative foetal fibronectin is not a useful test for inducibility at term. PMID- 11858890 TI - A population-based case-control teratologic study of nitrazepam, medazepam, tofisopam, alprazolum and clonazepam treatment during pregnancy. AB - OBJECTIVE: To study the association between nitrazepam, medazepam, tofisopam, alprazolum and clonazepam treatments during pregnancy and prevalence of different congenital abnormalities (CAs). MATERIALS AND METHODS: A matched case-control study using cases with CAs and population controls from the dataset of the nationwide Hungarian Case-Control Surveillance of Congenital Abnormalities (HCCSCA), 1980-1996. RESULTS: Of 38,151 pregnant women who had babies without any defects (population control group), 75 (0.20%) were treated with these five benzodiazepines during pregnancy. Of 22,865 pregnant women who delivered offspring with CAs, 57 (0.25%) had benzodiazepine treatment. The occurrence of five benzodiazepine treatments during the second and third months of gestation, i.e. in the critical period for most major CAs did not show significant differences in matched case-control pairs. CONCLUSION: Treatment with five benzodiazepines studied during pregnancy did not present detectable teratogenic risk to the fetus in humans but the amount of information was limited for different CAs. PMID- 11858891 TI - Production of human mast cell chymase in human myometrium and placenta in cases of normal pregnancy and preeclampsia. AB - OBJECTIVE: To investigate whether the human mast cell chymase-endothelin-1(1-31) system was present in human myometrium, chorion and umbilical cord in normal pregnancy. METHODS: Myometrium, placenta and umbilical cord were obtained from five normal pregnant women and 10 with preeclampsia. Each tissue was stained with antibodies against hMC and ET-1(1-31). RESULTS: Routine cells were located mainly around vessels. The number of hMC-positive cells and production of ET-1(1-31) were significantly higher in myometrium from patients with severe preeclampsia compared to those from normal pregnant women. In contrast, their numbers were significantly lower in placenta and umbilical cord in patients with severe preeclampsia. CONCLUSIONS: These results suggest that the hMC-ET-1(1-31) system is active in normal pregnancy. Overproduction of hMC and ET-1(1-31) in the myometrium may be involved in the pathogenesis of severe preeclampsia, and in such cases some defense mechanism may operate in the fetus to cope with the pathological effect of the hMC-ET-1(1-31) system. PMID- 11858892 TI - Trisomy 13 or 18 (mosaicism) in first trimester cytotrophoblast cells: false positive results in 11 out of 51 cases. AB - OBJECTIVE: The finding of full or mosaic trisomy 13 or 18 in first trimester chorionic villus sampling (CVS) may be a false-positive result. This report provides incidence and outcome information that may be helpful in counselling individual patients and in choosing adequate follow-up. STUDY DESIGN: From a series of 6820 CVS cases, we retrospectively collected data on all patients (n=51) with full (n=30) or mosaic (n=5) trisomy 18, and full (n=13) or mosaic (n=3) trisomy 13 in cytotrophoblast cells. RESULTS: Five false-positives were seen in patients with full trisomy 18 and three in the mosaic cases. One false positive result was observed in full trisomy 13 and two false-positives in cases of mosaicism. No false-negative results were reported. CONCLUSION: The diagnosis of trisomy 13 or 18 in cytotrophoblasts should be confirmed in other tissues, unless fetal abnormalities are seen at ultrasound. In case of mosaicism, follow up amniocentesis is advised. PMID- 11858893 TI - Vascular endothelial growth factor, interleukin-6 and interleukin-2 in serum and follicular fluid of patients with ovarian hyperstimulation syndrome. AB - BACKGROUND: The pathogenesis of ovarian hyperstimulation syndrome (OHSS) is not completely understood. OBJECTIVE: To investigate the presence of VEGF, IL-6 and IL-2, in serum and follicular fluid, in patients developing severe OHSS. STUDY DESIGN: We enrolled 101 women undergoing in vitro fertilization. Eight patients developing severe OHSS were compared with 43 high risk patients and 50 controls. We analyzed VEGF and IL-6 in serum collected before hCG administration, and in both serum and follicular fluid on the day of oocyte retrieval. RESULTS: OHSS patients presented follicular fluid IL-6 levels higher than both the patients at risk and controls (P<0.05). On the day of the oocyte retrieval the patients developing OHSS showed serum and follicular VEGF values higher than the ones of the patients at risk (P<0.05). Serum and follicular fluid IL-2 levels showed no differences between the examined groups. IL-2, IL-6 and VEGF values were not correlated with each other. CONCLUSIONS: Angiogenesis and inflammation processes are both present in severe OHSS. PMID- 11858894 TI - Seasonal variations in human seminal parameters. AB - OBJECTIVES: To obtain data on seasonal variations of sperm parameters in an andrology laboratory. STUDY DESIGN: Semen parameter values and hormone values of 2454 patients attending our infertility clinic between 1990 and 1997 were analysed. Seasonal trends were calculated using the method of Edwards [Ann. Hum. Genet. 25 (1961) 83]. RESULTS: The total group of mean sperm count did not show a significant variation, but a significant circannual trend occurred of patients born in the groups 1950-1954, 1955-1959 and 1965-1969. A significant variation of acrosin activity occurred with a maximum in March. The other parameters did no show significant variations. CONCLUSIONS: The knowledge on circannual variation of semen parameters and hormone values may be of value in diagnostic and therapeutic decisions in reproductive medicine. PMID- 11858895 TI - Management of ureteric injuries during gynecological operations: 10 years experience. AB - OBJECTIVE(S): To present our 10 years experience in the management of ureteric injuries occurring during gynecological surgery. STUDY DESIGN: Seventy-six patients had a variety of injuries. In 29 cases, the ureteric damage was diagnosed intraoperatively. Management of early-diagnosed injuries included suturing, ligature removal, end-to-end anastomosis, and reimplantation of the ureter. In 47 cases, the injury was diagnosed postoperatively. Ureteric catheterization was attempted in all cases presenting with obstruction. Catheterization failures were managed with ureterolysis, and reimplantation. Small ureteric fistula were managed with catheterization, and large communications with reimplantation. Two cases with urinomas were treated with surgical evacuation and anastomoses. RESULTS: Management of early-diagnosed injuries was relatively easy in most cases. Postoperatively-diagnosed injuries were more difficult to treat. Catheterization failed in 28/44 (65.9%) ureters and surgical re-exploration was necessary. Long-term morbidity was minimal and no relapses occurred. CONCLUSION(S): Early recognition of a ureteric injury is the key to a complications-free repair. Unrecognized injuries cause prolonged morbidity, and their management can be difficult. Treatment of these injuries by experienced teams may minimize long-term consequences. PMID- 11858896 TI - Factors predicting disease outcome in early stage adenocarcinoma of the uterine cervix. AB - CONTEXT: Adenocarcinoma (AC) and adenosquamous carcinoma (ASC) comprise the second principal histological types of cervical carcinoma. As compared with the squamous cell cancer (SCC), these lesions are far less frequent, and their epidemiology, natural history and prognostic determinants are less well understood. OBJECTIVE: Patients with an early stage AC of the uterine cervix diagnosed in our clinic were subjected to detailed analysis for the prognostic determinants. STUDY SUBJECTS: A series of 94 women with early stage (adenocarcinoma in situ (AIS) to IIB) cervical ACs or ASCs diagnosed and treated in our department during 1995-1999 and subsequently followed-up for a mean of 43.1 +/- 16.2 (S.D.) months. MAIN OUTCOME MEASURES: Patients were examined by colposcopy, Papanicolaou (PAP) smear and biopsy. The stage of the disease (FIGO) and tumour histology in operative specimens were recorded, and univariate (Kaplan Meier) and multivariate survival analysis (Cox) were run to explore the factors predicting disease outcome. RESULTS: Mean age of the women was 44.2 +/- 2.5 (S.D.) years (range 24-81 years), which is significantly (P=0.000) lower than that (49.9 +/- 14.2) of 464 SCC patients in our material. Minority of the women (38.2%) reported any clinical symptoms, but these correlated with the stage (P=0.041). Screening history was acceptable (i.e. screening interval 3 to 4 years) in 56 women, whereas 28 (29.8%) had no previous PAP smear taken. Interpretation errors were established in 17 (23.6%) and sampling errors in 6 (8.3%) of the 72 smears available for re-screening. No colposcopic lesions were found in 29 (30.9%) women. Follow-up data were available from 72 patients, of whom the disease progressed in four (one died), whereas 68 patients are alive and well at the moment. Patient's age (P=0.000), screening history (P=0.0127), FIGO stage (P=0.001), mode of therapy (P=0.0187), and presence of co-existent squamous cell lesions (P=0.0184) were significant prognostic indicators in univariate survival analysis. Cox's multivariate survival analysis disclosed FIGO stage (P=0.001) and screening history (P=0.006) as the only significant independent predictors of the disease outcome. CONCLUSIONS: The present data emphasise the importance of early cervical AC as a disease of younger women, making early detection of its precursors (AIS) by regular PAP smear screening mandatory in prevention of disease progression. This can only be achieved by increasing the sensitivity of the PAP smear in detecting abnormal glandular cells in asymptomatic women. PMID- 11858897 TI - Optimal surgery and diagnostic approach of stage IA2 squamous cell carcinoma of the cervix. AB - BACKGROUND: Most patients with International Federation of Gynecology and Obstetrics (FIGO) stage IA2 squamous cell carcinoma of the cervix, opt for radical surgery at present. OBJECTIVE: To review surgical and diagnostic approaches in such patients. STUDY DESIGN: Our patient population consisted of 394 patients with a diagnosis of stage I squamous cell cervical carcinoma (with depth of stromal invasion 10mm or less) according to the 1995 FIGO definition. Biopsy and surgical specimen slides were reassessed retrospectively in all cases. The findings of T2-weighted MR imaging were available from the individual medical charts. RESULTS: None of the patients with stromal invasion of 5mm depth or less showed pelvic lymph node metastasis. However, metastasis to the parametrial connective tissue was found in one case with stage IA1 exhibiting marked lymph vascular space involvement. There were no deaths due to disease in cases with stromal invasion of 5mm depth or less. The lesions were detected in all 20 cases exhibiting stromal invasion of greater than 5mm in depth. In contrast, the lesions were not detected with T2 imaging in four of six cases (67%) with stage IA2. CONCLUSION: Simple or modified radical hysterectomy with pelvic lymph node dissection may be sufficient for cases of stage IA2 cervical squamous cell carcinoma where lymph-vascular space involvement is absent. T2-weighted MR imaging with no detectable tumor would prove beneficial in the selection of these patients. PMID- 11858898 TI - Ovarian cancer in BRCA-positive women: vigilance is mandatory despite screening programs. AB - A young BRCA-1 positive woman turned out to have advanced stage ovarian cancer despite an ensuring check-up 4 months earlier. Much remains unknown regarding the efficacy of screening programs, when applied to individuals predisposed for an inherited malignancy. Therefore, vigilance is mandatory when dealing with women with a positive family history of mammarian and/or ovarian cancer. PMID- 11858899 TI - Vesical calculus causing dystocia. AB - Vesical calculus is a rare cause of dystocia. We report a case in a 22-year-old primigravida. PMID- 11858900 TI - Recurrent impetigo herpetiformis in a pregnant adolescent: case report. AB - Impetigo Herpetiformis is a rare pustular dermatosis that typically occurs in pregnant women with unknown etiology. A 17 year old patient who developed Impetigo Herpetiformis for the second time in the 27th week of her 2nd pregnancy is presented. The patient improved with corticosteroids treatment but the lesions did not clear completely and had flare ups during stressful periods which brings us to conclusion that Impetigo Herpetiformis at least has a common pathway with Generalized Pustular Psoriasis in the pathogenesis as stress provoked exacerbations. PMID- 11858901 TI - Endometrial mixed Mullerian tumor with heterologous elements following tamoxifen therapy for breast cancer: a case report and literature review. AB - CASE: A 67-year-old multiparous woman received 20mg tamoxifen daily for four years after surgical treatment of breast cancer. She presented with vaginal bleeding. Uterine curettage revealed a uterine MMT with heterologous elements. She was treated surgically with adjuvant radiotherapy. Tumor cells were found to be estrogen receptor negative and progesterone receptor positive. Uterine MMT may be linked to long term use of tamoxifen. A mechanism in developing MMT other than estrogen receptor pathway may be possible. PMID- 11858902 TI - A real-time PCR assay for the detection of Neisseria gonorrhoeae by LightCycler. AB - The detection of Neisseria gonorrhoeae by the polymerase chain reaction (PCR) is now recognized as a sensitive and specific method of diagnosing infection by the organism. In this Study 152 urine specimens were examined for N. gonorrhoeae by a real-time PCR method using the LightCycler platform and results were compared to an "in-house" PCR assay using an ELISA-based detection method. N. gonorrhoeae DNA was detected in 29 (19%) specimens by LightCycler PCR (LC-PCR) and in 31 (20%) specimens by the "in house" PCR method. The LightCycler assay proved to be specific and 94% sensitive when compared to the "in house" PCR method. These features combined with the rapid turn-around time for results makes the LC-PCR particularly suitable for the detection of N. gonorrhoeae in a routine clinical laboratory. PMID- 11858903 TI - Training costs and investment payback of implementing molecular diagnostics for identification of vancomycin resistant enterococci in a clinical microbiology laboratory. AB - Molecular diagnostics may be a more efficient method to manage resources; but most Microbiology laboratories have not introduced them into routine use due to the specialized training required. Using vancomycin resistant enterococci (VRE) screening during a comparison of a multiplex PCR (MPCR) and conventional biochemical testing (CBT) we studied 3 objectives: 1) to develop a molecular diagnostics in-house training program, 2) to assess the training program outcomes for competency and confidence, and 3) to determine laboratory payback. A training program for 14 technologists using multiple adult learning methods was implemented. Methods to minimize technical errors were introduced and included: use of a calibrated loop to deliver sample; prealiquotting reagents; increasing volume of specimen; addition of gel loading dye directly into reaction tubes; and establishment of an equivocal zone. In our laboratory MPCR costs $7.06 less than CBT, therefore the payback period for training and implementation would be approximately 3 years. PMID- 11858904 TI - Rapid diagnosis of enterovirus infections by real-time PCR on the LightCycler using the TaqMan format. AB - A 2 h single-tube, reverse-transcription(RT)-PCR/hybridization assay using the TaqMan format for rapid diagnostic screening of enterovirus (EV) infections was optimized for the real-time LightCycler (LC) technology. For low EV load clinical samples an additional 30 min reamplification step using a novel primer-mix/probe design resulted in a 100% concordance with AMPLICOR EV PCR Test and in-house RT PCR. Combined with maximum specificity, the sensitivity of LC-PCR was 10- to 100 fold higher compared to AMPLICOR EV Test. PMID- 11858905 TI - In vitro activity of the new ketolide ABT-773 against community acquired respiratory tract isolates and Viridans Streptococci. AB - A total of 230 isolates were collected from clinical specimens of patients attending five health centers of the Buenos Aires, Argentina. ABT-773 was compared to erythromycin, azithromycin and clindamycin against bacterial isolates responsible for community-acquired respiratory tract infections and viridans streptococci showing different resistance patterns. Time-kill curves were also performed against selected resistant isolates. All but one of the 105 pneumococcal isolates were susceptible to ABT-773. Among the erythromycin resistant S. pyogenes isolates, all the M type and inducible isolates were susceptible to ABT-773. ABT-773 showed excellent activity against macrolide, azalide, lincosamide (MAL) inducible S. aureus producers but was inactive against constitutive producers. ABT-773 activity against viridans streptococci was also excellent.ABT-773 exerted bactericidal activity against selected isolates of S. pneumoniae, M. catarrhalis and H. influenzae, however, it was only bacteriostatic against methicillin-susceptible S. aureus. PMID- 11858906 TI - In vitro activity of gemifloxacin compared to seven other oral antimicrobial agents against aerobic and anaerobic pathogens isolated from antral sinus puncture specimens from patients with sinusitis. AB - Using a microbroth method for aerobes and agar dilution for anaerobes, we studied the comparative in vitro activity of gemifloxacin, three fluoroquinolones, two macrolides and two beta-lactams, against 207 aerobic and 162 anaerobic antral sinus puncture isolates. Gemifloxacin was active at < or = 0.25 ug/mL against 198/207 [96%] aerobes and 127/162 [78%] anaerobes or 325/369 [88%] of all isolates and was the most active quinolone on a weight basis against Gram positive organisms. All Haemophilus and Moraxella species were susceptible to < or =0.06 ug/mL of gemifloxacin. Thirty-five anaerobic isolates [35/162, 22%] required > or =0.5 ug/mL of gemifloxacin for inhibition, including all Prevotella species [all except one strain of P. bivia were susceptible to < or = 2 ug/mL] and occasional strains of Bacteroides uniformis, Bacteroides fragilis, Bilophila wadsworthia, Peptostreptococcus magnus, Peptostreptococcus micros, Propionibacterium acnes, and Veillonella species. All fusobacteria were susceptible to < or =0.25 ug/mL of gemifloxacin. Based on our in vitro study results, we conclude that gemifloxacin may offer a therapeutic alternative for sinus infections. PMID- 11858907 TI - In vitro evaluation of AZD2563, a new oxazolidinone, tested against unusual gram positive species. AB - The recently introduced oxazolidinone, linezolid, has a spectrum and potency directed against Gram-positive organisms, including antimicrobial-resistant isolates. The newest agent in this class, AZD2563 was tested against uncommonly isolated Gram-positive species to establish the breadth of its spectrum. By reference broth microdilution methods, 120 strains were tested (48 Corynebacterium spp., 10 species; 27 Listeria spp., 2 species; 11 Micrococcus spp., 2 species; 23 Bacillus spp., 3 species; 6 Stomatococcus mucilaginosus and one strain each of 5 other species) against AZD2563 and compared to eight other agents. The AZD2563/linezolid MIC(50;) % inhibited at < or =4 microg/mL were: for corynebacteria (0.25/0.25 microg/mL; 100/100%), Listeria spp. (2/2 microg/mL; 100/100%), Micrococcus spp. (1/1 microg/mL; 100/100%), Bacillus spp. (0.5/1 microg/mL; 100/100%), and S mucilaginosus (0.5/1 microg/mL; 100/100%). Using the MIC(90) values, AZD2563 was slightly more potent than linezolid (two-fold). Only four genus- species groups had AZD2563 MICs of strains at 2 microg/mL (Aerococcus, Leuconostoc, Listeria, Rhodococcus), all other isolates were inhibited by < or = 1 microg/mL. The AZD2563 potency and spectrum versus these rarer species was at least equal to linezolid, including some strains resistant to penicillins, macrolides-lincosamides, and fluoroquinolones. PMID- 11858908 TI - In vitro activity of clinafloxacin in comparison with other quinolones against Stenotrophomonas maltophilia clinical isolates in the presence and absence of reserpine. AB - A total of 33 Stenotrophomonas maltophilia clinical isolates were tested for their susceptibility to clinafloxacin in comparison with ciprofloxacin, levofloxacin, moxifloxacin, nalidixic acid, norfloxacin, sparfloxacin and trovafloxacin. The MIC(50) and MIC(90) were as follows: ciprofloxacin 4 and 64 microg/mL; clinafoxacin 0.5 and 4 microg/mL; levofloxacin 2 and 32 microg/mL; moxifloxacin 1 and 8 microg/mL; nalidixic acid 8 and 128 microg/mL; norfloxacin 64 and 256 microg/mL; sparfloxacin 1 and 16 microg/mL; and trovafloxacin 1 and 8 microg/mL. Clinafloxacin was the most active quinolone, with only a 15.1% of strains showing resistance. When the MICs were determined in the presence of 25 microg/ml of reserpine, the MIC(90) of trovafloxacin and moxifloxacin did not change, whereas decreased 2-fold for clinafloxacin, levofloxacin, sparfloxacin and nalidixic acid, and 4- and 8-fold for ciprofloxacin and norfloxacin respectively. No clinafloxacin-resistant strains were observed when the MIC was performed in the presence of reserpine. Therefore, clinafloxacin shows the better "in vitro"activity against these 33 strains of S.maltophilia. PMID- 11858909 TI - Antimicrobial susceptibilities of Streptococcus pyogenes isolated from respiratory and skin and soft tissue infections: United States LIBRA surveillance data from 1999. AB - This study evaluated current levels of antimicrobial resistance and associated demographic trends among clinical isolates of Streptococcus pyogenes in the United States as part of the LIBRA surveillance initiative. In 1999, 2,742 isolates of S. pyogenes (2,039 respiratory; 405 skin and soft tissue; 148 blood) were collected from 324 clinical laboratories and centrally tested for antimicrobial susceptibility by the broth microdilution method. All isolates were susceptible to penicillin (MIC(90,) < or = 0.06 microg/mL), ceftriaxone (MIC(90,) < or =0.03 microg/mL), vancomycin (MIC(90,) 0.5 microg/mL), levofloxacin (MIC(90,) 1 microg/mL), and moxifloxacin (MIC(90,) 0.25 microg/mL). Twenty-four (0.9%) azithromycin-intermediate (MIC, 1 microg/mL) and 170 (6.2%) azithromycin resistant (MIC, > or = 2 microg/mL) isolates were identified. Regionally, azithromycin resistance varied by < 5%, ranging from 3.0% in New England to 7.7% in the Pacific region. Azithromycin resistance was significantly higher (P < 0.001) among patients aged 15-64 years (8.3%) than patients < or =14 years (4.3%). This study found higher rates of macrolide resistance among S. pyogenes than previously reported in the United States and suggests that macrolide resistance is common among respiratory isolates from both younger and older patients. Fluoroquinolones (moxifloxacin > levofloxacin) demonstrated potent in vitro activity against all isolates of S. pyogenes tested, including those from skin and soft tissue infections. Given the higher rates of macrolide resistance reported in other countries and the seriousness of invasive infections, continued antimicrobial surveillance of S. pyogenes in the United States would be prudent. PMID- 11858910 TI - Linezolid-resistant Enterococcus faecium isolated from a patient without prior exposure to an oxazolidinone: report from the SENTRY Antimicrobial Surveillance Program. AB - A patient case report describes an Enterococcus faecium strain isolated from a blood culture that was resistant to linezolid (MIC, 8 microg/mL; G2576U mutation of 23S rRNA). Co-resistances were identified for vancomycin, ampicillin, macrolides, fluoroquinolones, chloramphenicol, rifampin, gentamicin (high-level), nitrofurantoin and trimethoprim/sulfamethoxazole. Etest (AB BIODISK, Solna, Sweden) and disk diffusion results also detected the oxazolidinone resistance pattern. Laboratories should be aware of the rare possibility of these strains occurring during linezolid therapy or spontaneously (this case) in contemporary practice, and have in vitro susceptibility methods available capable of detecting oxazolidinone resistance. PMID- 11858911 TI - Leg ulcer due to Pseudomanas luteola in a patient with sickle cell disease. AB - Pseudomonas luteola is rarely implicated in clinical infections, usually in association with indwelling catheters or prostheses. This report describes the first case of deteriorated leg ulcer caused by a multiply antibiotic resistant P. luteola strain in a patient with homozygous sickle cell disease. PMID- 11858912 TI - In vitro activity of 11 antimicrobial agents, including gatifloxacin and GAR936, tested against clinical isolates of Mycobacterium marinum. AB - In vitro activities of 11 antimicrobial agents were determined using Etest (AB BIODISK, Solna, Sweden) against 37 clinical isolates of Mycobacterium marinum. The most potent agent (MIC(50,) 0.047 microg/mL) was trimethoprim/sulfamethoxazole, but only 91.9% would be categorized as susceptible. Several agents had a complete spectrum of activity including gatifloxacin (MIC(90,) 0.38 microg/mL), minocycline (MIC(90,) 2 microg/mL), tetracycline (MIC(90,) 2 microg/mL) and amikacin (MIC(90,) 3 microg/mL). The new glycylcycline, GAR936, had a similar potency (MIC(90,) 3 microg/mL) to that of the parent minocycline compound (MIC(90,) 2 microg/mL), but the range of MIC values extended to 24 microg/mL. Numerous options appear to exist for the contemporary therapy of M. marinum infections including some newer fluoroquinolones and derivatives of tetracycline (doxycycline, minocycline, GAR936). PMID- 11858913 TI - Clindamycin resistant strains of Clostridium difficile isolated from cases of C. difficile associated diarrhea (CDAD) in a hospital in Sweden. AB - Fifty three strains of C. difficile recovered from the stools of 13 patients with clinical C. difficile associated diarrhea (CDAD) were analyzed for the presence of the ermB gene, for toxigenicity and fingerprinting profile by PCR based assays. Forty five percent of the isolates were resistant to clindamycin and positive for the ermB gene. All clindamycin resistant isolates were ermB positive and belonged to the same fingerprinting group, suggesting clonal spread. These preliminary results suggest that clindamycin resistant isolates may be common etiologic agents of CDAD in Sweden. PMID- 11858914 TI - Enhanced diagnosis of narrow complex tachycardias with increased electrocardiograph speed. AB - The objective of this study was to determine if the addition of rapid speed (50 mm/s) electrocardiograms (EKGs) improves the clinician's diagnostic accuracy of narrow complex tachycardias when compared to standard speed (25 mm/s) EKGs. We conducted a prospective, comparative trial. Forty-five difficult narrow complex tachycardias (heart rate range: 150-250 beats per minute) were printed at both 25 mm/s and 50 mm/s. Eight board certified emergency physicians initially interpreted the standard speed EKG (standard group) and chose a therapy for the hypothetical patient. These eight participants later interpreted the same 45 EKGs by using both the standard and rapid speed EKGs (rapid group) and again chose a therapy. The gold standard for each EKG was based on the patient's clinical diagnosis and was independently confirmed in all cases by a board-certified cardiologist. The rhythm distribution was as follows: atrial flutter (17), atrial fibrillation (11), paroxysmal supraventricular tachycardia (15), and sinus tachycardia (2). Participants were masked to all clinical information and EKG ratios. Diagnostic accuracy was compared by using McNemar's chi(2) test. Correct diagnosis improved from 226/360 (63%), in the standard group to 257/360 (71%) in the rapid group (difference in means 8.6%, p = 0.002). The incorrect use of adenosine was decreased from 43/240 (18%) in the standard group to 32/240 (13%) in the rapid group (difference in means 4.5%, p = 0.06). In conclusion, correct diagnosis of difficult narrow complex tachycardias was improved when EKGs at both 25 mm/s and 50 mm/s were used for interpretation. It appears that the simple technique of increasing the EKG paper speed, and thus effectively spacing out the rhythm, enhances the diagnostic ability of the observer. PMID- 11858915 TI - EMS activation in a cohort of critically ill patients. AB - The objective of this study was to examine Emergency Medical Services (EMS) activation in a cohort of critically ill patients and to evaluate whether factors such as perceived difficulty of access to emergency care, insurance status, and educational level are associated with EMS use. Patients admitted from the emergency department to the intensive care unit (ICU) of an urban county hospital for diagnoses other than acute cardiac conditions or stroke were interviewed about EMS use. Overall EMS use was found to be 52%. Primary reasons for not activating EMS were a belief that symptoms were not serious (36%) or that self transport was faster (25%). The only factors significantly associated with bypassing EMS were car ownership, lack of health insurance, and self-decision to go to an emergency department. Sixty-eight percent of the sample reported little difficulty in accessing emergency care. In conclusion, because of a belief that symptoms were not serious or self-transport would be faster, nearly half of the critically ill patient sample did not activate EMS. Patient education about warning sign recognition for serious illnesses and about the potential benefits derived from EMS should be improved. PMID- 11858916 TI - Prospective randomized trial of heliox-driven continuous nebulizers in the treatment of asthma in the emergency department. AB - The purpose of this study was to evaluate the efficacy of heliox-driven continuous nebulizers in the management of moderate to severe asthma exacerbations in the Emergency Department (ED). The trial was a prospective, randomized, double-blind, controlled trial in a university Emergency Department (volume 65,000 patients/year) of patients 18-55 years of age with acute asthma exacerbations. Patients were placed on continuous nebulizers driven by 70:30 heliox or air with 30% O(2). Respiratory rate, peak expiratory flow rate (PEFR), forced expiratory volume at 1 s (FEV1), and Borg dyspnea scale were measured at 0 and 2 h. Disposition and patient satisfaction were also assessed. A total of 36 patients completed the study (18 heliox, 18 air). There was no difference between groups in baseline variables. There was significant interval improvement within both groups at 2 h in PEFR, FEV1, Borg, and respiratory rate. There was no significant difference between heliox 70:30 and air/oxygen in PEFR [mean between group improvement difference 17 liters/min, 95% confidence interval (CI) -20-51], FEV1 (0.03 liters/sec, 95% CI -0.22-0.30), or respiratory rate (mean between group improvement difference 0.5, 95% CI -2.7-3.8). There was a significant improvement in the patients' perceived dyspnea as measured by the Borg dyspnea scale at 2 h in the heliox group (1.6, 95% CI 0.3-3.0). In this trial of patients with moderate-severe asthma exacerbation, heliox-driven continuous nebulizers failed to demonstrate an improvement in respiratory rate, oxygen saturation, PEFR, or FEV1 at 2 h. However, there was a significant improvement in the patients' perceived dyspnea on heliox over air/oxygen measured by the Borg dyspnea scale. PMID- 11858917 TI - Road traffic accident mortality in Singapore. AB - The aim of this study was to identify factors that contribute to road traffic accident mortality and the patterns of injuries sustained by these victims, with a view to identifying areas for future intervention. All road traffic accident deaths that occurred in Singapore over a period of 1 year were reviewed. A total of 226 deaths occurred: 82.3% of the victims were male. The median age was 31 years. Blood alcohol was detected in 42 (18.7%) victims. In general, head (86.7%), followed by thoracic (67.7%) and abdominal (31.4%) injuries, were the most common injuries. Severe lower extremity trauma was most common among pedestrians and pedal cyclists (20.6% and 11.0%, respectively). The mean Injury Severity Score was 38.7. The relative risk of mortality between motorcyclists and motorcar drivers was 18.8:1. Suggestions for future prevention and intervention include stricter enforcement of speed limits, more severe penalties for drunk driving, helmet use among pedal cyclists, and the introduction of pre-hospital advanced airway management. PMID- 11858918 TI - Improved identification of acute coronary syndromes with second generation cardiac troponin I assay: utility of 2-hour delta cTnI > or = +0.02 ng/mL. AB - It has been shown that the 2-h Stratus II delta creatine kinase-MB (CK-MB) is more sensitive and is equally specific compared to a 2-h Stratus II CK-MB and to a 2-h Stratus II delta cardiac troponin-I (DeltacTnI) for identification of acute myocardial infarction and adverse outcome (AO). Because the newest generation of Stratus (Stratus CS) cTnI assay has an analytical sensitivity of 0.03 ng/mL, compared to 0.35 ng/mL for the first generation assay, we undertook a small pilot study of 120 chest pain patients to compare sensitivities and specificities for 30-day AO of the Stratus CS DeltacTnI immunoassay to the DeltaCK-MB and DeltacTnI, as measured by the Abbott Axsym immunoassay, and to the DeltaCK-MB, as measured by the Stratus CS. A Stratus CS DeltacTnI > or = +0.02 ng/mL in 2 h was more sensitive (61.9%) than an Axsym DeltaCK-MB > or = +1.3 ng/mL (38.1%; p = 0.03), a Stratus CS DeltaCK-MB > or = +0.4 ng/mL (38.1%; p = 0.03), and an Axsym DeltacTnI > or = +0.3 ng/mL (33.3%; p = 0.03) for 30-day AO. There were no differences in specificities. Our data support enhanced identification of ACS with a second generation cTnI assay. Pending larger studies, patients with a rise in DeltacTnI of > or = +0.02 ng/mL in 2 h, as measured by the Stratus CS immunoassay, should receive consideration for aggressive anti-ischemic therapy and further diagnostic testing prior to making an exclusionary diagnosis of non ischemic chest pain. PMID- 11858919 TI - A comparison of oral, tympanic, and rectal temperature measurement in the elderly. AB - This cross-sectional study enrolled 95 elderly Emergency Department patients aged 60 years or more to determine if rectal temperatures identify fevers more often than oral or tympanic temperatures when the chief complaint suggests an infection. A fever was defined as a temperature greater than 38 degrees C (100.4 degrees F). Discordance was defined as any patient with an oral or tympanic temperature of 38 degrees C or less but manifesting a rectal temperature greater than 38 degrees C and 0.5 degrees C (1 degrees F) greater than the oral or tympanic temperature. Rectal thermometry identified a fever in 14 of 95 (14.7%) patients who were afebrile orally and in 11 of 90 (12.2%) patients who were afebrile tympanically. Five of 90 (5.6%) patients were febrile rectally but were afebrile by both oral and tympanic thermometry. Thus, rectal thermometry identified fevers missed orally and tympanically in elderly patients whose presentation suggested infection. To identify these febrile patients with possible infection, clinicians must be attentive to elderly patients' vague clinical presentation. PMID- 11858920 TI - Potential drug-drug interactions in elderly patients presenting with syncope. AB - Adverse drug reactions and interactions are a common cause of iatrogenic disease. Although there are resources available that have been developed for assessing potential drug-drug interactions (DDIs), one must first consider the possibility in order to apply these tools. This cross-sectional analysis of elderly patients evaluates Emergency Physician documentation of potential DDIs and assesses the utility of computer software recognition of interactions that can lead to syncope. In this study, lack of documentation of a patient's past medical history and medications suggests that Emergency Physicians are not evaluating DDIs as a possible source of their patient's symptoms. The computer resources evaluated were found to be easy to use and reliable. Rectifying and preventing potential DDIs can be an indispensible service provided by the Emergency Physician. PMID- 11858921 TI - Digital foot trauma: emergency diagnosis and treatment. AB - Trauma to the foot is commonly seen in the emergency setting. An accurate diagnosis and step-wise management is appropriate. Although most injuries respond to aggressive non-surgical treatment, surgical intervention may be required for an unstable or a displaced fracture, particularly of the first metatarsophalangeal joint. PMID- 11858922 TI - Painless acute aortic dissection and rupture presenting as syncope. AB - Acute aortic dissection is an emergency that may not only cause significant morbidity but often results in death. A timely diagnosis can prove difficult in the event of an atypical presentation. Classically, aortic dissection presents as sudden, severe chest, back, or abdominal pain that is characterized as ripping or tearing in nature. This article reports on a case of a completely painless acute dissection and rupture in a middle-aged man who presented with syncope. The diagnosis was made by a combination of clinical suspicion, emergent bedside echocardiography, and computed tomography scan. This patient underwent immediate surgery, and ultimately had a successful outcome. PMID- 11858923 TI - Chronic meningitis presenting with acute obstructive hydrocephalus. AB - A previously healthy 24-year-old woman presented to the Emergency Department unresponsive with a Glasgow Coma Scale score of 4 and evidence of brainstem herniation. She was intubated and hyperventilated. Computed axial tomography scan of the brain demonstrated four-chamber hydrocephalus. Continued hyperventilation and mannitol diuresis were sufficient to arrest the impending herniation while emergent ventriculostomy was arranged. The patient recovered without sequelae and ultimately received a diagnosis of chronic idiopathic meningitis. This case highlights a rarely diagnosed disorder that presented with an acutely life threatening condition. PMID- 11858924 TI - Subarachnoid hemorrhage and isolated atresia of the aortic arch. AB - The case of a 14 year old boy with subarachnoid hemorrhage and atresia of the aorta without patent ductus arteriosus or intracardiac shunt is described. This case calls attention to the possibility of aortic obstruction in adolescents or young adults with hypertensive stroke. The clinical symptoms, radiographic findings, and surgical repair of isolated aortic interruption, including atresia, are discussed. PMID- 11858925 TI - Hypermagnesemia-induced fatality following epsom salt gargles(1). AB - Hypermagnesemia is a rare cause of coma in a patient with normal renal function. When present, it is often because of iatrogenic medication overdose. We report a fatal case of chronic Epsom salt gargles for halitosis that produced a serum magnesium of 23.6 mg/dL (9.8 mmol/L) and resulted in coma. We review the wide presentation of hypermagnesemia from subtle neurologic and cardiovascular signs to the major life-threatening clinical manifestations of shock, dysrhythmias, coma, and cardiopulmonary arrest despite emergency dialysis. PMID- 11858926 TI - Electrocardiographic manifestations: right ventricular infarction. AB - The 12-lead electrocardiogram (EKG) is an essential tool when evaluating the Emergency Department (ED) patient with suspected cardiac ischemia. The standard EKG has limitations when evaluating "remote" areas of the heart such as the left posterior wall or right ventricular wall. Diagnosis of right ventricular infarction (RVI) in the presence of acute inferior wall myocardial infarction (MI) is made utilizing right-sided chest leads with high sensitivities and specificities. RVI is a serious ED problem because morbidity and mortality is higher in acute MIs associated with RVI. PMID- 11858927 TI - Adenosine dose should be less when administered through a central line. AB - A patient with a port-a-cath was given 12 mg of adenosine for paroxysmal supraventricular tachycardia (PSVT), resulting in prolonged (13 s) bradycardia and severe side effects. When the same patient presented 2 weeks later for recurrent PSVT, only 3 mg of adenosine was needed to terminate the episode, without the patient experiencing prolonged bradycardia or severe side effects. The literature suggests that for patients with central venous catheters, a lower dose of adenosine should be used to terminate PSVT. PMID- 11858928 TI - A snake bite associated with trauma. PMID- 11858929 TI - Late rebound digoxin toxicity after digoxin-specific antibody Fab fragments therapy in anuric patient. PMID- 11858930 TI - Blebs associated with rattlesnake envenomation followed by serum sickness. PMID- 11858931 TI - Illusive gunshot wound to the chest. PMID- 11858932 TI - Longus colli tendinitis causing acute neck pain. PMID- 11858933 TI - What is cherry red, and who cares? PMID- 11858936 TI - PTEN: The down side of PI 3-kinase signalling. AB - The PTEN tumour suppressor protein is a phosphoinositide 3-phosphatase that, by metabolising phosphatidylinositol 3,4,5-trisphosphate (PtdIns(3,4,5)P(3)), acts in direct antagonism to growth factor stimulated PI 3-kinases. A wealth of data has now illuminated pathways that can be controlled by PTEN through PtdIns(3,4,5)P(3), some of which, when deregulated, give a selective advantage to tumour cells. Early studies of PTEN showed that its activity was able to promote cell cycle arrest and apoptosis and inhibit cell motility, but more recent data have identified other functional consequences of PTEN action, such as effects on the regulation of angiogenesis. The structure of PTEN includes several features not seen in related protein phosphatases, which adapt the enzyme to act efficiently as a lipid phosphatase, including a C2 domain tightly associated with the phosphatase domain, and a broader and deeper active site pocket. Several pieces of data indicate that PTEN is a principal regulator of the cellular levels of PtdIns(3,4,5)P(3), but work is only just beginning to uncover mechanisms by which the cellular activity of PTEN can be controlled. There also remains the vexing question of whether any of PTEN's cellular functions reflect its evolutionary roots as a member of the protein tyrosine phosphatase superfamily. PMID- 11858937 TI - G protein-coupled receptor interacting proteins: emerging roles in localization and signal transduction. AB - The mechanism by which G protein-coupled receptors (GPCRs) translate extracellular signals into cellular changes initially was envisioned as a simple linear model: activation of the receptor by agonist binding leads to dissociation of the heterotrimeric GTP-binding G protein into its alpha and betagamma subunits, both of which can activate or inhibit various downstream effector molecules. The plethora of recently described multidomain scaffolding proteins and accessory/chaperone molecules that interact with GPCR, including GPCR themselves as homo- or heterodimers, provides for diverse molecular mechanisms for ligand recognition, signalling specificity, and receptor trafficking. This review will summarize the recently described GPCR-interacting proteins and their individual functional roles, as understood. Implicit in the search for the functional relevance of these interactions is the expectation that enhancement or disruption of target cell-specific events could serve as highly selective therapeutic opportunities. PMID- 11858938 TI - Leukemia inhibitory factor enhances bFGF-induced IL-6 synthesis in osteoblasts: involvement of JAK2/STAT3. AB - We previously showed that basic fibroblast growth factor (bFGF) stimulates release of vascular endothelial growth factor (VEGF) and synthesis of interleukin 6 (IL-6) in osteoblast-like MC3T3-E1 cells. In the present study, we investigated the effects of leukemia inhibitory factor (LIF) on the release of VEGF and IL-6 in these cells. LIF did not affect the bFGF-stimulated VEGF release. On the contrary, LIF, which alone had little effect on IL-6 release, significantly enhanced the bFGF-stimulated IL-6 release. The amplifying effect of LIF on the IL 6 release was dose dependent in the range between 0.01 and 10 ng/ml. AG490, an inhibitor of JAK2, suppressed the amplifying effect of LIF. LIF induced the phosphorylation of STAT3. AG490 inhibited the LIF-induced STAT3 phosphorylation. Taken together, our results strongly suggest that LIF enhances bFGF-stimulated IL 6 synthesis via JAK2/STAT3 pathway in osteoblasts. PMID- 11858939 TI - Interleukin-7 receptor expression and activation in nonhaematopoietic neoplastic cell lines. AB - The high-affinity human interleukin-7 (IL-7)R is a heterodimeric complex consisting of the IL-7Ralpha and common interleukin-2 receptor gamma (IL 2Rgamma(c)) chains. Activation of the IL-7R complex is associated with tyrosine and serine residue phosphorylation of a number of intracellular substrates leading to proliferation and induction of various cellular differentiation processes. In this study, we demonstrate, by S1 nuclease protection assay, immunoprecipitation and in vitro kinase assay that functional human (h) IL-7R is expressed in haematopoietic and nonhaematopoietic cell lines. The National Cancer Institute (NCI) tumour panel of 60 cell lines (NCI60) was screened for the expression of IL-7R mRNA by S1 nuclease protection assay, and IL-7R mRNA was detected in 9 of 12 leukemia, 3 of 7 lung, 4 of 6 CNS, 2 of 7 melanoma, 2 of 7 renal, 1 of 6 colon and 1 of 6 breast cancer cell lines. Immunoblot analysis of haematopoietic, lung cancer and brain tumour cell lines demonstrated expression of IL-7R, IL-2Rgamma(c) and p59 fyn, suggesting that the components of an IL-7R signalling network are present in nonhaematopoietic neoplastic cells. Immunoprecipitation of IL-7Ralpha followed by an in vitro kinase assay demonstrated functional receptor phosphorylation events in the lung cancer cells but not in the brain tumour cell lines. The expression of functional IL-7R on epithelial tumour cells may represent a potential target for receptor-directed therapy. PMID- 11858940 TI - Cadmium-induced DNA synthesis and cell proliferation in macrophages: the role of intracellular calcium and signal transduction mechanisms. AB - Cd(2+) exposure increases the risk of cancer in humans and animals. In this report, we have studied the effect of Cd(2+) on signal transduction and Ca(2+) mobilization in murine macrophages. At micromolar concentrations, Cd(2+) significantly increased cell division as judged by [3H]thymidine uptake and cell counts. Cd(2+)-treated cells continued to proliferate even after more than 4 weeks in culture. Cd(2+) (1 microM) treatment induced a 1.5- to 2-fold increase in cytosolic free Ca(2+), [Ca(2+)](i), which was transitory and/or oscillatory. The sources of this Ca(2+) included both inositol 1,4,5-trisphosphate (IP(3)) sensitive and -insensitive stores. Macrophage treatment with 1-(6-((17beta-3 methoxyestra-1,2,5(10)-triene-17-yl)amino)hexyl)-1H-pyrrole-2,5-dione (U73122), an inhibitor of phosphatidylinositol-specific phospholipase C (PLC), decreased Cd(2+)-induced formation of IP(3) in a concentration-dependent manner (K(d) about 2 microM). This caused a concomitant, partial decrease in the effect of Cd(2+) on [Ca(2+)](i). Cd(2+) itself crosses the macrophage membrane in part via L-type Ca(2+) channels, but it also interacts with a cell surface membrane protein(s) coupled to a pertussis toxin-sensitive G protein. Use of selective inhibitors of signal transduction and the quantitation of the levels of phosphorylated MAPK/ERK activating kinase-1 (MEK1), extracellular signal-regulated kinase-1 (ERK1), and p38 mitogen-activated protein kinase (MAPK) suggests that the effects of Cd(2+) are mediated by the p21(ras)-dependent MAPK, but not the phosphoinositide 3 (PI 3)-kinase signalling pathway. The effect of activating these pathways includes increased availability of the transcription factor NFkappaB as well as activation of the early genes c-fos and c-myc. PMID- 11858941 TI - Differential involvement of Src family kinases in pervanadate-mediated responses in rat myometrial cells. AB - We previously described that pervanadate, a potent tyrosine phosphatase inhibitor, induced contraction of rat myometrium via phospholipase (PL) C-gamma1 activation [Biol Reprod 54 (1996) 1383]. In this study, we found that pervanadate induced tyrosine phosphorylation of the platelet-derived growth factor (PDGF) beta receptor, interaction of the phosphorylated PDGF receptor with the phosphorylated PLC-gamma1, production of inositol phosphates (InsPs), extracellular signal-regulated kinase (ERK) activation and DNA synthesis. All these responses were insensitive to PDGF receptor kinase inhibition or PDGF receptor down-regulation. We showed that Src family kinases were activated by pervanadate, and that InsPs production and phosphorylation of both PLC-gamma1 and the PDGF receptor were blocked by PP1, an Src inhibitor. In contrast, the stimulation of ERK by pervanadate was totally refractory to PP1. These results demonstrated that the activation of Src by pervanadate is involved in PLC gamma1/InsPs signalling but does not play a major role in ERK activation. PMID- 11858942 TI - Peroxisome proliferators activate growth regulatory pathways largely via peroxisome proliferator-activated receptor alpha-independent mechanisms. AB - Peroxisome proliferators (PPs) induce liver tumors in rodents through an unknown mechanism requiring PP-activated receptor (PPAR) alpha. Since PPs possess growth modulatory activities that may be important to their hepatocarcinogenicity, we aimed at dissociating the activation of growth signaling pathways from the PPARalpha-mediated response induced by PPs in cultured rat primary hepatocytes. Pretreatment with the differentiation-promoting agent dimethylsulfoxide (DMSO) increased PPARalpha mRNA/protein and enhanced the expression of PPARalpha regulated genes [fatty acyl Co-A oxidase (FACO), cytochrome P450 4A1 (CYP4A1)] induced by PPs. In contrast, DMSO reduced the expression of immediate early genes (IEG) expression (c-myc, c-jun, c-fos, junB, egr-1) and inhibited mitogen activated protein kinase (MEK) kinase/extracellular signal-regulated kinases (ERKs) and p38 phosphorylation. Furthermore, the inhibitors Tyrphostin and PD98059 dowregulated IEG/ERKs induction and slightly enhanced the FACO/CYP4A1 response induced by the PP WY-14,643. The stimulation of signal transduction pathways by PPs can be dissociated from PPARalpha activation, thus suggesting that PPs could activate growth regulatory pathways largely via PPARalpha independent mechanisms. PMID- 11858943 TI - Nerve growth factor activates kinases that phosphorylate atypical protein kinase C. AB - Activation of atypical protein kinase C by nerve growth factor (NGF) involves phosphorylation. In order to identify kinases that regulate atypical PKC (aPKC), we surveyed PC12 cell lysates for protein kinases that are activated by NGF and which could phosphorylate aPKC. Employing an in-gel kinase assay where aPKC-zeta was copolymerized within the gel matrix as a substrate, three kinases, pp175, pp87 and pp60, were identified as enzymes that phosphorylated aPKC. Phosphorylation of aPKC by these three kinases coincided with NGF-induced activation of the enzyme. Each kinase possessed a unique subcellular distribution pattern and could be activated by either ceramide or H(2)0(2), second messengers that mimic NGF signaling events. Upstream, pp175 and pp60 lie in a ras pathway, whereas pp87 lies in a pathway dependent upon src. Altogether, these findings reveal that the aPKCs are subject to regulation by a novel group of kinases. PMID- 11858944 TI - Signal transduction mechanism via adenosine A1 receptor in the cat esophageal smooth muscle cells. AB - We investigated what adenosine receptor type exists and the signaling pathways on the contraction of circular muscle cells isolated by enzymatic digestion from the cat esophagus. Adenosine or the selective A1 receptor agonist R-PIA causes a concentration-dependent contraction. After pretreatment with A1 receptor antagonist, DPCPX, adenosine-mediated contraction was abolished. Adenosine induced contraction was significantly increased when A1 receptors were preserved by pretreatment with DPCPX followed by inactivation of all unprotected receptors with N-ethylmaleimide. Adenosine- or R-PIA-induced contraction was significantly augmented in the preserved cells and the increase was abolished in the presence of the A1 receptor antagonist DPCPX. PTX abolished contraction induced by adenosine or R-PIA, implying that contraction activated by A1 receptor was coupled to a pertussis toxin (PTX)-sensitive G(i) protein. After permeabilization, contraction was inhibited by G(i2), but not by G(i1) and G(i3), antibodies. These data suggest that adenosine-induced contraction of esophagus depends on PTX-sensitive G(i2.) Adenosine- or R-PIA-induced contraction of esophageal smooth muscle cells was not affected by the phospholipase D (PLD) inhibitor rho-chloromercuribenzoic acid (rhoCMB), phospholipase A(2) (PLA(2)) inhibitor DEDA or PKC antagonist chelerythrine, but was significantly abolished by phospholipase C (PLC) inhibitor, neomycin. PLC-beta3 antibody inhibited R-PIA induced contraction. R-PIA-induced contraction of esophageal muscle cells was inhibited by IP(3) receptor antagonist heparin, which suggests that the contraction of esophageal smooth muscle cells is dependent on phosphatidylinositol-specific phospholipase (PI-PLC) and IP(3). In conclusion, adenosine- and R-PIA-induced contraction in cat esophageal smooth muscle cell was mediated by A1 receptor. A1 receptor is coupled to PTX-sensitive G protein G(i2), which results in the activation of PI-PLC-beta3. PI hydrolysis by PI-PLC forms IP(3), which binds to IP(3) receptor on endoplasmic reticulum, resulting in the release of intracellular Ca(2+). PMID- 11858945 TI - Choline phosphate potentiates sphingosine-1-phosphate-induced Raf-1 kinase activation dependent of Ras--phosphatidylinositol-3-kinase pathway. AB - In NIH3T3 cells, sphingosine-1-phosphate (S1P) caused a significant increase of Raf-1 kinase activity as early as 2 min. Interestingly, choline phosphate (ChoP) produced synergistic increase of S1P-stimulated Raf-1 kinase activation in the presence of ATP while showing additive effect in the absence of ATP. However, Raf 1 kinase activation induced by S1P decreased in the presence of ATP when applied alone. The overexpression of N-terminal fragment of Raf-1 (RfI) to inhibit Raf- Ras interaction caused the inhibition of S1P-induced Raf-1 kinase activation. Also, wortmannin, phosphatidylinositol-3-kinase (PI3K) inhibitor, exhibited inhibitory effects on S1P-induced activation of Raf-1 kinase. In addition, we demonstrated that the chemical antioxidant, N-acetylcysteine attenuated Raf-1 activation induced by S1P, suggesting that H(2)O(2) may be required for the signalling pathway leading to Raf-1 activation. This H(2)O(2)-induced Raf-1 kinase activation was also blocked by inhibition of Ras--PI3K signalling pathway using alpha-hydroxyfarnesylphosphonic acid and wortmannin. Taken together, these results indicate that S1P-induced Raf-1 kinase activation is mediated by H(2)O(2) stimulation of Ras--PI3K pathway, and is enhanced by ChoP in the presence of ATP. PMID- 11858946 TI - Itch--mediators and mechanisms. AB - A specialized subpopulation of unmyelinated chemonociceptors and dedicated spinal neurons which are responsible for the itch sensation have been identified recently. Under physiological conditions, painful stimuli such as activation of conventional mechano-heat-sensitive ('polymodal') nociceptors (scratching) inhibit the itch sensation via central mechanisms. Conversely, centrally acting pain-inhibiting opioids enhance itch by disinhibition. These mechanisms might well explain the itch in diseases characterized by histamine release like urticaria, and might provide evidence for the role of endogenous opioids as central itch promotors in cholestasis or nephropathy. After the discovery of itch specific neurons has dramatically improved our understanding of itch mechanisms under experimental conditions, the present task is to correlate these new findings to the clinical situation of itch patients. PMID- 11858947 TI - Effect of tap-water iontophoresis on sweat gland recruitment, skin temperature and skin blood flow. AB - Our interest was to quantify the role of sweat gland activation on the maintenance of skin temperature during mild exercise in the heat. Seven days of tap-water iontophoresis decreased the number of active sweat glands by 72% which significantly increased forearm skin temperature and blood flow during mild exercise (70 W) in the heat (32 degrees C). Skin temperature of the treated forearm was 0.5 degrees C warmer (P=0.049); skin blood flow in the treated forearm was 13% higher than the control arm (P=0.021). These results illustrate the importance of sweat evaporation on skin temperature and blood flow during exercise. PMID- 11858948 TI - A novel mutation of the tyrosinase gene causing oculocutaneous albinism type 1 (OCA1). AB - Tyrosinase is a rate-limiting enzyme in the melanin biosynthetic pathway and a complete defect of the enzyme activity caused by homozygous mutations of the tyrosinase gene is well known to result in tyrosinase-negative oculocutaneous albinism (OCA1A) patients who never develop any melanin pigment in the skin, hair and eyes throughout life. In this paper, we report a novel missense substitution, R239W(CGG --> TGG) of the tyrosinase gene in a patient with tyrosinase-negative OCA. PMID- 11858949 TI - Cell death and proliferation in Nd-YAG laser, electrocautery, and scalpel wounds on mice skin. AB - The purpose of this study is to compare cell death and proliferation in laser, electrocautery and scalpel wounds on the mice epidermis. Wounds were examined by transmission electron microscopy, the detection of free 3'-OH DNA ends and immunohistochemistry of proliferating cell nuclear antigen (PCNA), inducible nitric oxide synthase (iNOS), keratinocyte growth factor (KGF) and keratinocyte growth factor receptor (KGFR). Reepithelization was first observed 5 days after scalpel and laser incisions and 7 days after electrocautery incision. Ultrastructurally, keratinocytes in both electrocautery and laser wounds showed similar post-apoptotic necrotic changes. Interestingly, dividing cells were often observed 3 days after laser incision. Apoptotic index in electrocautery wounds was higher than in laser wounds, although there was no significant difference in the PCNA expression level between them. The expression of iNOS, KGF and KGFR in laser wounds was more intense than in electrocautery wounds. In scalpel wounds, keratinocytes did not show significant changes in morphology or of markers of cell death and proliferation during the observation period. Therefore, the increase in the number of dividing cells and in the expression level of iNOS, KGF and KGFR may induce earlier and thicker reepithelization in laser wounds than in electrocautery and scalpel wounds. PMID- 11858950 TI - Immunohistochemical study of membrane type-matrix metalloproteinases (MT-MMPs) and matrix metalloproteinase-2 (MMP-2) in dermatofibroma and malignant fibrous histiocytoma. AB - Matrix metalloproteinases (MMPs) play an important role in tumor invasion and metastasis. Enhanced expression of matrix metalloproteinase-2 (MMP-2) has been demonstrated in dermatofibroma (DF) and malignant fibrous histiocytoma (MFH). MMP 2 has been shown to be activated by membrane-type MMPs (MT-MMPs). To study the role of MT-MMP in the activation of MMP-2, skin specimens of DF (five cases) and MFH (three cases) were immunohistochemically studied using in situ zymography and the antibodies against matrix metalloproteinase-2 (MMP-2) and membrane type 1-3 MMPs (MT1-3-MMPs). Both MMP-2 activity and its expression were significantly activated in the tumor cells in DF and MFH. Anti-MT2-MMP strongly reacted with tumor cells of all cases of DF and MFH, whereas anti-MT1 or 3-MMP antibody showed a weak reaction in some cases of DF and MFH. Double immunofluorescence labeling demonstrated that the immunoreactive cells with anti-MMP-2 antibody in DF and MFH consistently reacted with anti-MT2-MMP antibody. The results suggest that the activation of MMP-2 in the benign and malignant fibrous tumors is related to the activation of MT-MMPs. PMID- 11858951 TI - Expression of cellular prion-related protein by murine Langerhans cells and keratinocytes. AB - Transmissible spongiform encephalopathies are characterized by the accumulation of a proteinase-resistant isoform of the cellular prion-related protein (PrP(c)) within the central nervous system (CNS). The accumulation of scrapie-associated PrP (PrP(Sc)) within cells of the lymphoreticular system prior to its accumulation in the CNS is regarded as important for the development of neurological diseases after peripheral inoculation. Little, however, is known as to which cells are the targets for peripheral inoculation. Here, the presence of PrP(c) on murine Langerhans cells (LC), dendritic cells in the skin and mucosa, and keratinocytes (KC) is demonstrated by immunohistochemical staining, Western blotting and FACS analysis. The expression of PrP(c) mRNA in freshly purified LC and KC was also detected by reverse transcriptase-polymerase chain reaction. The expression of PrP(c) on LC was slightly increased during culture. These data suggest that LC and KC may be the targets for peripheral infection with prions. PMID- 11858952 TI - Increased synthesis of calcitonin gene-related peptide stimulates keratinocyte proliferation in murine UVB-irradiated skin. AB - Repeated ultraviolet (UV) irradiations have been shown to induce keratinocyte proliferation with acanthosis, stimulate the cutaneous nerve proliferation, and increase the synthesis of calcitonin gene-related peptide (CGRP). In the current study, we examined the role of CGRP in the UVB-induced proliferation of murine keratinocytes. UVB irradiation increased the number of bromodeoxyuridine (BrdU) labeled basal keratinocytes and caused acanthosis. In addition, CGRP expression was up-regulated in the peripheral nerves of the upper dermis and lower epidermis. Repeated intradermal injections of CGRP increased the number of BrdU labeled basal cells and caused acanthosis. Intradermal injections of capsaicin prior to UVB-irradiation inhibited the UVB-induced CGRP expression, BrdU labeling in basal keratinocytes and epidermal thickening. Intradermal injections of anti CGRP antibody inhibited the UVB-induced BrdU labeling in basal keratinocytes, but epidermal thickening was not significantly inhibited. These results indicate that CGRP is one of the stimulators to UVB-induced keratinocyte proliferation. On the other hand, expression of substance P, another neuropeptide in the peripheral nerve, was not up-regulated by UVB irradiation. PMID- 11858953 TI - Impaired repair ability of hsp70.1 KO mouse after UVB irradiation. AB - UV light is absorbed in the epidermis and induces sunburn cell formation. It has been reported that HSP70 increases the UVB resistance of cell lines by in vitro experiments using various cell lines. In this study, hsp70.1(-/-) KO mouse was used in order to study the role of HSP70 after UVB irradiation. Western blotting showed a decreased level of HSP70 in hsp70.1(-/-) KO mouse compared with wild type FVB mouse. Six h after UVB irradiation, there were no significant histologic differences between the hsp70.1(-/-) KO mouse and the wild type FVB mouse. A similar degree of nuclear swelling was observed. However, there were significant differences at 12 and 24 h after UVB irradiation. After 12 h, a few apoptotic cells were observed in the wild type mouse, but a large number of cells were apoptotic in the hsp70.1(-/-) KO mouse. After 24 h, the epidermis of the wild type FVB mouse was relatively intact, but almost the entire epidermis was necrotic in the hsp70.1(-/-) KO mouse. These results showed that epidermal injury of hsp70.1(-/-) KO mouse was much more severe than that of wild type mouse although initial changes are similar in both species of mice. These results suggest that susceptibility of hsp70.1(-/-) KO mouse to UVB irradiation may originate from a defect in the repair mechanism. This HSP deficient model may be useful in studies of the effects of tissue injury that relate to the impaired tissue repair mechanisms. PMID- 11858954 TI - Sustained ability for fibroblast outgrowth from stored neonatal foreskin: a model for studying mechanisms of fibroblast outgrowth. AB - Fibroblast cultures derived from neonatal foreskin explants have been an important model for understanding the basic mechanisms of fibroblast function and activity. Neonatal foreskin has been routinely used for establishing such fibroblast cultures in vitro. In general, tissue explants and fibroblast cultures are established from freshly harvested neonatal foreskin tissue. It is unknown whether prolonged storage of tissue, even in optimal growth media and conditions would still result in successful explant outgrowth. Specifically, no studies have properly examined the maximal duration of foreskin storage that can produce viable fibroblasts with normal growth and synthetic characteristics. We, therefore, set out to isolate and propagate fibroblast cultures from neonatal foreskin tissue that had been kept in complete culture media at 4 degrees C for various periods of time. We found that outgrowth and propagation of viable fibroblasts in vitro occurred with foreskin samples obtained within 24 days after surgical harvesting. The morphology, cellular proliferative capacity (measured by [3H]-thymidine uptake), steady state levels of alpha1(I) procollagen mRNA, and collagenous protein synthesis were comparable among fibroblast cultures derived from foreskin explants established 3 days (control) and up to 24 days (but not 38 days) after tissue harvesting. These studies demonstrate that viable and functional fibroblasts, with apparently similar in vitro characteristics, can be isolated and propagated from explants of neonatal foreskin tissue that have undergone prolonged storage. Moreover, these findings may be useful in understanding the lack of fibroblast growth in such conditions as found in delayed wound healing and aging. PMID- 11858955 TI - Nuclear factor kappaB (NF-kappaB) activation by hydrogen peroxide in human epidermal keratinocytes and the restorative effect of interleukin-10. AB - The heterodimeric form of nuclear factor kappaB (NF-kappaB), NF-kappaB1/RelA, is one of the pluripotential transcription factors that activates various genes encoding cytokines and cell adhesion molecules. To clarify the involvement of radical oxygen species in the NF-kappaB activation pathway in keratinocytes, we examined the effect of hydrogen peroxide (H(2)O(2)) on the activation of NF kappaB in cultured normal human epidermal keratinocytes. After the treatment of keratinocytes with 300 microM H(2)O(2), a translocation of NF-kappaB from the cytoplasm to nucleus was observed in an immunofluorescence study using anti-human NF-kappaB1 and anti-human RelA antibodies. Specific DNA binding was observed with the nuclear extract prepared from the H(2)O(2)-treated keratinocytes by the electrophoretic mobility shift assay. The presence of N-acetyl-L-cysteine or pyrrolidine dithiocarbamate during H(2)O(2) treatment prevented the nuclear localization of NF-kappaB. The involvement of radical oxygen species in the NF kappaB activation pathway was suggested. Pretreatment of keratinocytes with 10 ng/ml of recombinant human interleukin-10 (IL-10) for 24 h suppressed the nuclear translocation of NF-kappaB induced by H(2)O(2). IL-10, which increases in ultraviolet-irradiated skin and suppresses delayed type hypersensitivity in vivo, may play an inhibitory role in cutaneous inflammation by inhibiting the NF-kappaB activation pathway in keratinocytes. PMID- 11858956 TI - Abstracts of the 3rd European College of Neuropsychopharmacology Workshop. Nice, France, March 15-17, 2002. PMID- 11858957 TI - 40 years of the EORTC: the evolution towards a unique network to develop new standards of cancer care. European Organisation for Research and Treatment of Cancer. PMID- 11858958 TI - The EORTC Laboratory Research Division. European Organisation for Research and Treatment of Cancer. AB - The Laboratory Research Division (LRD) of the EORTC currently consists of five Groups with expertise that includes pre-clinical drug development, all aspects of cancer pharmacology, clinically-relevant receptor and biomarker studies, functional imaging and contemporary pathology. The LRD provides a Europewide resource for cancer clinical trials with particular expertise in the evolving field of translational research. In the development of therapies designed to exploit the molecular and cellular pathology of cancer, it is essential that translational research is included at all stages and the EORTC, through the LRD, has access to such expertise. In addition to providing support for drug development and clinical trials, the LRD represents a unique forum for the development of contemporary translational research expertise, the establishment of quality standards and the education of young laboratory and clinical scientists embarking on careers in oncology. PMID- 11858959 TI - The EORTC and drug development. European Organisation for Research and Treatment of Cancer. AB - Early drug development at EORTC has always been subject to structural changes to adapt to the rapid changes that occur in oncological drug development. The expertise of early drug developers has always been cross-fertilised with disease /tumour-oriented groups and also backwards to the laboratory research groups. This results in the establishment of a solid and dedicated network of medical oncologists with focused expertise in cancer drug development. The EORTC Data Center is fully equipped with all expertise to support clinical research activities and includes regulatory, safety, and quality assurance desks. The EORTC New Drug development Programme (NDDP) provides methodological expertise to early clinical trials and coordinates phase I and phase II studies addressing various approaches. Through NDDP, the early clinical groups and the disease /tumour-oriented groups have created specific networks to address early drug development in specific tumour types. This results in very efficient networks which have the resources and the patients to address and conduct challenging clinical trials in a standardised fashion ensuring the highest standards in cancer treatment. PMID- 11858960 TI - Scientific review of EORTC trials: the functioning of the New Treatment Committee and Protocol Review Committee. AB - New anticancer treatments (new therapeutic strategies or new compounds) require careful development in which cancer clinical trials are an essential element. Two scientific committees, namely the New Treatment Committee and the Protocol Review Committee, ensure the review of all EORTC protocols with respect to the interest and originality, methodology, feasibility and relevance within the EORTC framework. Both Committees are involved early in the evaluation of the new concept proposal and follow all aspects (methodology, administrative, regulatory) of the protocol development process. Throughout its 25 years of existence, the Protocol Review Committee has streamlined drug and protocol evaluations and has developed standard operating procedures to handle those reviews in a very efficient and fast manner. Since 1997, the New Treatment Committee has contributed to strengthening the EORTC during the development process with the aim of ensuring an optimal flow of information on new drugs between laboratory and clinical research divisions. PMID- 11858961 TI - The EORTC Boron Neutron Capture Therapy (BNCT) Group: achievements and future projects. AB - Boron Neutron Capture Therapy (BNCT) is an experimental treatment modality that takes place in a nuclear research reactor. To progress from preclinical studies to patient treatment is a challenge requiring strict quality management and special solutions to licensing, liability, insurance, responsibility and logistics. The European Organisation for the Research and Treatment of Cancer (EORTC) BNCT group has started the first European clinical trial of BNCT for glioblastoma patients at the European High Flux Reactor (HFR) in Petten, The Netherlands, conducted by the Department of Radiotherapy of the University of Essen, Germany. A very strict quality management had to be installed following the European rules on safety and quality assurance for nuclear research reactors, for radioprotection, for radiotherapy and for clinical trials. The EORTC BNCT Group has created a virtual European-wide hospital to handle the complex management of patients treated with BNCT. New clinical trials are currently under development. PMID- 11858962 TI - EORTC Brain Tumor Group: achievements and perspectives. AB - The EORTC Brain Tumor Group has a long standing history of achievements. The activities of the Brain Tumor Group have recently been re-structured in order to face the challenge of large intergroup/intercontinental trials and be a reference address for early drug development needed for gliomas. Constant adaptation to higher quality assurance criteria and implementation of translational research studies are now priorities for the Brain Tumor Group. Due to such activities, the EORTC Brain Group has become a major player in clinical research. The number of centres and patients joining its trials have greatly increased over the past 2 years. Achievements and strategies are detailed in this article. PMID- 11858963 TI - The EORTC Breast Cancer Group: 40 years of research contributing to improve breast cancer management. AB - The EORTC Breast Cancer Group (EBCG) is a multidisciplinary international group created in 1962 in conjunction with the EORTC. This group has always focused its activities on the development of new cancer treatments and strategies for all categories of breast cancer. It has been active both in drug development, as well as in the development of new radiotherapy and surgical techniques to attempt to improve cure rates and loco-regional control. Over 40 years, the EBCG has performed dozens of clinical studies including several thousands of patients. Many of these studies have contributed to the clinical knowledge on the treatment of breast cancer and have influenced the standard management of this tumour. Beside its clinical research activities, the Group has also been very active in developing guidelines for breast cancer research, promoting high standards of care in conferences, as well as in fellowships and quality assurance programmes. EBCG is a founding member of the Breast InterGroup (BIG). PMID- 11858964 TI - The Children Leukemia Group: 30 years of research and achievements. AB - The EORTC Children Leukemia Group (CLG) is part of the offspring of the EORTC Hemopathies Working Party which in 1978 split into a paediatric and to an 'adult' branch. At that time, the Berlin-Frankfurt-Munster (BFM) designed by H. Riehm for acute lymphoblastic leukaemia (ALL) appeared much more efficacious than all others and the CLG decided to adapt that treatment strategy for its own clinical trials. The main results of these may be summarised as follows:for standard risk patients, the deletion of cyclophosphamide from consolidation and reconsolidation courses does not jeopardise the patient's outcomefor medium- and high-risk patients receiving high-dose methotrexate (MTX), cranial radiotherapy is superfluouswith the dose scheduling of the BFM regimen, E-Coli L-Asparaginase is more efficacious than Erwinia L-Asparaginasethe addition of monthly intravenous (i.v.) 6-mercaptopurine to conventional maintenance chemotherapy is detrimentalthe assessment by quantitative polymerase chain reaction (PCR) of minimal residual disease at completion of induction is feasible in a cooperative setting and can be used as a powerful and independent prognostic factor. The CLG also conducted clinical studies of acute myeloblastic leukaemia. Since 1989, lymphoblastic non-Hodgkin's lymphomas have been treated within the ALL trials. The CLG collaborates with other Groups within the I-BFM Study Group and participants in the meta-analytic studies conducted by the Oxford team by the Oxford Children ALL Collaborative Group. PMID- 11858965 TI - It's time for chronotherapy! AB - The EORTC Chronotherapy Group (CTG) stemmed from the International Organisation for Cancer Chronotherapy (IOCC) in 1996. The IOCC was first to initiate large scale multicentre international chronotherapy trials, for the purpose of investigating the relevance of chronomodulated or timed administration of cancer therapy based on biological rhythms. Programmable pumps for cytotoxic chronodelivery and actigraph devices to monitor circadian rhythm alterations linked to cancer were also developed. The unique expertise of the IOCC with regard to cancer chronotherapy furthered its development within the EORTC. The EORTC offers broad expertise in clinical cancer research and opportunities for scientific recognition, intergroup collaborations and translational research. Over the past 5 years, the EORTC CTG has grown from 16 to 48 centres in 12 different countries. It is currently conducting seven multicentre chronotherapy trials which test the relevance of adapting cancer treatment delivery to circadian rhythms. The group aims at developing multiple collaborations to establish a chronotherapy network involving institutions with expertise ranging from experimental chronobiology to new drug testing, disease-specific management and quality of life or survival issues. PMID- 11858966 TI - The EORTC clinical research coordinators group. European Organisation for Research and Treatment of Cancer. AB - The Clinical Research Coordinators Group (CRCG) is an umbrella organisation, compiled from four existing groups, namely the Oncology Nurses Group, the Data Management Group, the Radiation Technologists Group and the Early Clinical Studies Group Research Nurses. From the existing steering committees, a new board was formed and consists of two members per group. The CRCG will function as an independent group within the EORTC. The CRCG will create conditions and standards for implementing and conducting clinical protocols according to Good Clinical Practice. PMID- 11858967 TI - EORTC cutaneous lymphoma task force. European Organisation for Research and Treatment of Cancer. AB - The Cutaneous Lymphoma Task Force has represented the European Organisation for Research and Treatment of Cancer (EORTC) over the last two decades and has received worldwide acceptance and the highest respect. The group has been able to bring together the world's experts in this field to try to solve the basic problems associated with primary lymphomas of the skin and to create a productive scientific research basis. The definition and the classification of the disease per se has been a major controversial problem and the development of an EORTC classification for primary cutaneous lymphoma has been one of the main goals of the group. The purpose of this paper is to highlight and to provide a historical perspective regarding the contribution of the EORTC Cutaneous Lymphoma Group to the development of consensus guidelines for securing uniform diagnosis, classification and management of the heterogeneous group of primary cutaneous lymphomas. Some future perspectives and strategies of the group are also presented. PMID- 11858968 TI - The EORTC Gastrointestinal Tract Cancer Group: 40 years of research contributing to improved GI cancer management. AB - The Gastrointestinal (GI) Tract Cancer Group's aims are to develop protocols concerning the different aspects of gastrointestinal tract malignancies, diagnosis, biology and mainly treatment. Prior to approval, new project proposals are discussed within different committees of the group and in the presence of specialists concerned, according to the type of trial. There are three main committees in the GI Group, chemotherapy, surgery and research. All projects of GI tract cancer are discussed first in the relevant committee before being discussed in the multidisciplinary plenary scientific session of the Group meeting. Multidisciplinarity has always been one of the major principles of the Group. This is well illustrated by the fact that the Chairman of the Group has been alternately a medical oncologist and a surgeon and is presently a surgeon. Radiation therapists also participate in the activity of the group. Like other EORTC groups, the GI Group has developed high standards of quality. Officers and members work in close co-operation with the staff of the Data Center in Brussels and in particular medical advisors, statisticians and data managers. Members from 32 different countries participate in the activities of the Group, mostly from European countries, but also from Russia, Egypt, Hong Kong, Israel, Peru, Russia, South Africa and many others through intergroup activities such as Australia, Canada and the USA. PMID- 11858969 TI - The EORTC Genito-Urinary Tract Cancer Group: 25 years of achievements and future strategies. AB - The European Organisation for Research and Treatment of Cancer (EORTC) Genito Urinary (GU) Tract Cancer Group celebrates 25 years of activity in 2001. The Group has developed an intense research activity carrying out phase II and phase III clinical trials in prostate, bladder, renal, penile and testicular cancers. It is one of the most active groups within the EORTC, entering more than 1200 new patients in its trials in 2001. In its trials, the EORTC GU Group also focuses on quality control, quality of life and uro-pathology. Besides collaboration with other EORTC groups, the GU Group is very actively collaborating with international organisations. Currently, several large phase III studies are conducted in collaboration with European and North American organisations. For the next few years, the Group is committed to develop projects aimed at testing new drugs and therapeutic strategies and increasing the collaboration between basic science and clinical practice. PMID- 11858970 TI - From chemoprevention and organ preservation programmes to postoperative management: major achievements and strategies of the EORTC Head and Neck Cancer Group. AB - The purpose of this article is to review the most significant results of the clinical studies conducted in the past two decades by the EORTC Head and Neck Cancer Group (HNCG). As for phase III trials, the HNCG investigated, besides the efficacy of chemopreventive drugs, the impact of cytostatic agents on various therapeutic targets, in combination or not with surgery and chemotherapy. These targets were: (a) chemo-prevention in curatively treated early stage disease; (b) organ preservation programmes in patients with operable tumour, comparing immediate surgery versus first-line chemotherapy; (c) postoperative management of locally advanced tumours, comparing radio-chemotherapy versus radiotherapy alone. Other phase II and phase III studies were also completed to investigate drug activity in advanced and/or recurrent head and neck squamous cell and adenoid cystic carcinomas. The present article will also analyse the strategies developed within the Group in the field of translational research. PMID- 11858971 TI - Management of infection in cancer patients. studies of the EORTC International Antimicrobial Therapy Group (IATG). AB - Infection remains an important cause of morbidity and mortality in cancer patients, especially those undergoing chemotherapy for haematological malignancies. The practice of instituting an empirical broad-spectrum antibiotic therapy as soon as possible after the onset of fever has substantially reduced the clinical impact of this complication. In the last 25 years, the International Antimicrobial Therapy Group of the European Organisation for Research and Treatment of Cancer (EORTC-IATG) have published nearly 30 articles and a number of abstracts on several facets of the epidemiology and management of infection in cancer patients. With a progressive methodological refinement, the EORTC-IATG trials have established the standard for the management of febrile neutropenia, both by setting methodologies and definitions and by testing several antibiotic regimens that are active and effective for this indication. With the aim of supporting a more rational use of antibiotics in cancer patients, the most recent trials have dealt with the management of low risk patients, showing the safety and feasibility of oral therapy. PMID- 11858973 TI - Highlights and strategies of the EORTC Leukemia Group. European Organisation for Research and Treatment of Cancer. AB - The EORTC Leukemia Group comprises more than 45 qualified haematology centres in Europe. The group promotes cooperation with new centres from Central and Eastern Europe with the aim to improve the standard and quality to the level in Western Europe. Subcommittees on cytogenetics, molecular biology, and immunology, shared by experts in the field, are active and have meetings on a twice-yearly basis. The aim of our group is to organise phase II and phase III trials for patients with acute and chronic myeloid or lymphoid leukaemia, myelodysplastic syndromes and myeloma. In 2000, 330 patients have been included in nine trials of the group. Presently, more than 2600 patients included in previous and current studies are alive and under continuous follow-up allowing studies on the long term results to be planned. PMID- 11858972 TI - Achievements and goals of the EORTC Invasive Fungal Infections Group. European Organisation for Research and Treatment of Cancer. AB - Invasive fungal infections are an increasing complication for patients with cancer. These infections still are difficult to diagnose and to treat and thus still have a high fatality rate. New strategies should include evaluation of new diagnosis tools and large-scale assessment of these new methods will need multidisciplinary collaboration. High-quality clinical trials dedicated to establish 'state-of-the-art' prevention and treatment are also directly needed. Created in 1991, the EORTC Invasive Fungal Infection Group has faced several of these challenges and significantly improved the knowledge and management of these infections in Europe. PMID- 11858974 TI - Overview of past, present and future of the EORTC Lung Cancer Group. European Organisation for Research and Treatment of Cancer. AB - The EORTC Lung Cancer Group (LCG) is a multidisciplinary international group of experts performing clinical research in lung cancer since 1962. Originally, the group consisted mainly out of French and Belgian investigators and expanded gradually into a wide range of investigators from all European Union countries, as well as some investigators from Switzerland, Poland, Czech Republic, Egypt, Slovenia, South Africa, Peru, Brazil and Cyprus. Despite the wide collaboration, it remains a difficult task to perform high quality large clinical research trials to answer important scientific questions in the treatment of lung cancer. For this reason, the EORTC Lung Cancer Group has invested a lot of efforts in promoting worldwide, randomised phase III studies in collaboration with other Groups. Furthermore, the LCG promotes small phase II trials of new drugs or treatments for lung cancer and stimulates the investigation of new strategies and treatments for rare intrathoracic malignancies. PMID- 11858975 TI - The achievements of the EORTC Lymphoma Group. European Organisation for Research and Treatment of Cancer. AB - From 1964 onwards, the EORTC Lymphoma Group has conducted seven consecutive randomised phase 3 trials on early stage Hodgkin's lymphoma aiming at increasing efficacy, while decreasing short- and long-term toxicity. Staging laparotomy is definitely abandoned and replaced by identification of prognostic subgroups based on pretreatment clinical characteristics. Event-free and overall survival significantly improved from about 50 and then 70%, in the early years, to over 80 and then 90% more recently. Radiotherapy fields have become more restricted, whereas chemotherapy has become standard. Longitudinal quality-of-life assessment has become an integral part of our studies. In advanced stages, overall outcome has improved as well with 6-year survival rates of over 80%. In aggressive types of NHL, the second generation chemotherapy schedule CHVmP-BV was superior to CHVmP. We could not show any advantage for intensification of upfront treatment with autologous stem cell transplantation. PMID- 11858976 TI - The EORTC Melanoma Group: a comprehensive melanoma research programme by clinicians and scientists. European Organisation for Research and Treatment of Cancer. AB - The EORTC Melanoma Group (MG) was founded in 1969 by both clinicians and scientists from various disciplines and fields of research with a common interest in malignant melanoma. This collaborative approach has always been the foundation of the groups strength. With an interest in tumour biology and especially the immunological aspects of the disease, the group has always pursued a scientific approach to treatment development in malignant melanoma. Over the years, the group has performed many clinical trials, epidemiological studies, histopathological studies defining standards and guidelines, translational research regarding prognostic factors and various metastatic and immunological aspects of melanoma, and developed quality assurance programmes for immunological and molecular biological assays in laboratory networks. At present, the EORTC MG runs the worldwide largest clinical trial programme in stages II, III and IV melanoma involving some 140 cancer centres in and outside Europe. Each trial is associated with the appropriate translational research programmes. PMID- 11858977 TI - The role of the EORTC pathologist in clinical trials: achievements and perspectives. European Organisation for Research and Treatment of Cancer. AB - The role of the pathologist in clinical trials (CT) is focused on three activities: pathology review, translational research, and participation in scientific committees. The primary goal of pathology review in CT is the quality control (QC) of the diagnosis and prognostic parameters. Important contributions have been achieved in the context of QC for CT such as new classifications of diseases or identification of new prognostic markers that are now widely used. Telematics implemented in some EORTC groups markedly facilitate the pathology review. The pathologist has a key-role in translational research for the identification of new targets in tissue specimens that may eventually lead to new therapeutics and for the understanding of the mechanisms involved in tumour progression. The gap between individualised prognosis and therapeutical possibilities has been considerably reduced by the development of drugs targeted on specific molecular defects. The paradigm of this is the treatment of stromal tumours by STI-571. For proper selection of patients to be treated, information on the expression of the molecules involved is needed, which is well suited for pathologists. The access to tissue resources from patients included in CT is a major goal to enhance translational research, both for brand institution and CT organisations. Active involvement of pathologists in scientific committees and interactions with the pharmaceutical industry is mandatory for an optimal design of CT protocols. In addition, translational research is a resource-consuming activity that necessitates an adequate financial flow to create a proper infrastructure at least for sponsored trials to the participating pathology departments and committees. PMID- 11858978 TI - Quality of life research within the EORTC-the EORTC QLQ-C30. European Organisation for Research and Treatment of Cancer. AB - In forming its Quality of Life Group, the EORTC created one of the earliest and largest of such groups in Europe. The EORTC QLQ-C30 which this group developed has become the most widely used questionnaire in Europe for cancer patients, and is extensively used around the world. The Quality of Life Group continues to build upon this success, both by refining the QLQ-C30, whilst developing a range of additional modules, and by initiating research projects that explore aspects of quality of life assessment, evaluation and interpretation. We review the progress to date and indicate directions of further research and development. PMID- 11858979 TI - EORTC Radiotherapy Group: achievements and future projects. European Organisation for Research and Treatment of Cancer. AB - The European Organisation for Research and Treatment of Cancer (EORTC) Radiotherapy (RT) Group will celebrate 27 years of activity in 2002. During its long history, the Radiotherapy Group has conducted a large number of studies which have provided valuable information on the radiation treatment of several disease sites. Group efforts have been concentrated on dose-effect studies, optimal fractionation schemes, combinations with other treatment modalities, and new radiotherapy techniques. The EORTC RT Group was the first in Europe to develop and introduce methodologies of Quality Assurance in radiotherapy. The RT Group actively collaborates with other EORTC Groups and international organisations. Currently, several phase III studies are being conducted in collaboration with European, North American and Australian organisations. The collaboration with RTOG led to the setting up of common systems for scoring late normal tissue effects. In the years to come, the Group will keep pioneering pivotal trials in radiotherapy and radio-chemotherapy. It will also explore combinations with novel therapies in phase I trials and implement innovative translational research programmes. PMID- 11858980 TI - The EORTC soft tissue and bone sarcoma group. European Organisation for Research and Treatment of Cancer. AB - The EORTC Soft Tissue Sarcoma Group was founded 25 years ago and has since developed into one of the leading cooperative groups in the research of sarcomas and has members from 40 institutions from 13 countries. The activities of the group have primarily been within the areas of standards for local as well as systemic treatment strategies, new drug development and quality control procedures. The group has a extensive quality control programme involving a strict membership policy, central review of the responses, central review of pathology, use a systemic therapy check-list and on-site monitoring of studies. A large database with over 2500 patients included in EORTC STBSG chemotherapy trials has been developed. So far, the STBSG has conducted more than 40 clinical trials accruing more than 250 patients per year, some of which has been performed in collaboration with other prestigious groups. PMID- 11858981 TI - Regulatory issues for clinical trials at EORTC: the way forward. European Organisation for Research and Treatment of Cancer. AB - Since the Declaration of Helsinki, the performance of clinical trials is subject to ethical and gradually also legal requirements. As the EORTC is performing clinical trials in more then 30 countries, it gained expertise in the field of Regulatory Affairs of all those countries. This paper intends to address the general approach with regard to European Regulatory Affairs. Furthermore, it is focusing on the role of the EORTC at the level of the competent regulatory authorities. As recently things are moving and changing on a European level, it describes the perception of the current and future European regulatory framework, and last, but not least, it is explained in what respect the achievements of the EORTC could be of benefit to society when defining their cancer treatment policies. PMID- 11858982 TI - Health Economics Unit achievements. AB - This article describes the activities of the EORTC Health Economics Unit since its inception in early 1994. The aim of the unit is to carry out economic evaluations of competing treatment options in common cancers in order to provide health care decision makers with useful information about the relative benefits and costs of the therapies they have to choose between. These assessments are mainly carried out by integrating collection of economic data in selected phase III randomized controlled clinical trials conducted by the EORTC collaborative groups. Studies with an economic evaluation integrated are currently becoming mature enough for analysis and several publications have resulted or are in press. Some studies with data from other sources than EORTC trials have also been performed and published. In addition, the unit has actively followed and contributed to the continuous methodological development in the field of economic evaluation of health care interventions. PMID- 11858983 TI - Quality assurance within EORTC: past, present and future. Introduction. European Organisation for Research and Treatment of Cancer. PMID- 11858984 TI - Quality assurance in medical oncology within the EORTC. European Organisation for Research and Treatment of Cancer. AB - Quality assurance (QA) lately arrived in the medical arena in comparison to other fields. EORTC focused initially its attention on the aspects related to clinical trial data handling. In the late 1980s, the EORTC appointed a Quality Control Committee (QCC) with the remit to expand the QA activities to the main disciplines involved in cancer treatment. From 1990 to 1996, two projects supported by the European Commission enabled the QCC to address, amongst others, specific questions related to medical oncology. Both projects focused on the practices of chemotherapy delivery and the quality of data reporting. Following these projects, the QCC developed standard guidelines to advise on chemotherapy delivery and also a systemic chemotherapy checklist to enable an easy collection of essential data onto the patient's files. More recently, the EORTC QA Committee proposed a minimal set of quality control procedures to be implemented by all EORTC groups. PMID- 11858985 TI - Quality assurance in radiotherapy: from radiation physics to patient- and trial oriented control procedures. AB - The stepwise process of the EORTC Quality Assurance Programme in Radiotherapy is described in function of two main criteria: the targets of the quality control procedures implemented, in Radiation Physics and clinical research, by the EORTC Radiotherapy Group and the development of both trial- and patient-oriented quality systems. This exhaustive program, which started in 1982, is characterised by three main periods. The first one was fully dedicated to pioneer steps in Radiation Physics measurements, on-site audits and inventories of human resources, staff workload and department infrastructure in institutions participating to EORTC trials. During the second period, which started in the late 1980s, a series of quality systems were implemented to test the compliance of the investigators to follow protocol guidelines, through the use of standard and uniform control procedures like the dummy runs, in order to tackle systematic errors in the participating institutions. Finally, the third period, which took place in the 1990s, was essentially patient-oriented, thanks to large scale individual case reviews, to check the validity of data recording and reporting processes and trace random errors throughout the radiotherapy treatments. Most of the results collected during these two decades allowed the implementation of well codified quality control procedures which, nowadays, can be used outside the field of clinical research, by national societies or bodies willing to improve treatment standards on a large scale. PMID- 11858986 TI - Quality assurance in EORTC clinical trials. European Organisation for Research and Treatment of Cancer. AB - Quality assurance (QA) has become a major factor over recent years in the management and analysis of clinical trials. The EORTC recognised very early the importance of QA in clinical data handling and started in 1992 with the development of the first SOP in the format of a 'Procedures manual' which, from the beginning, had incorporated the EU GCP guidelines. In 1995, a Quality Assurance Unit (QAU) was created to coordinate the various QA activities and to guarantee that all clinical trials do comply at all levels with a minimum of QA requirements. The QAU coordinates internal and external audits and is a mandatory partner in the audits performed by national/international authorities and pharmaceutical industries. This process has been prolonged at the Data Center with the development of a full set of standard operating procedures (SOPs), the implementation of training programmes for each category of staff and an ongoing interval monitoring process. PMID- 11858987 TI - Statistical methodology of phase III cancer clinical trials: advances and future perspectives. AB - The methodology for conducting cancer clinical trials has undergone enormous changes over the past 25-30 years since the EORTC Data Center was created. The purpose of this paper is to highlight and to provide a historical perspective for the main methodological concepts, both practical and theoretical, which form the basis for the design and analysis of phase III cancer clinical trials within the EORTC Data Center. Some statistical aspects of other associated topics such as quality of life, health economics, meta-analysis and treatment outcome will also be briefly discussed. Finally, some future perspectives and topics for further statistical methodological research will be presented in order to spur statisticians to meet the challenge of efficiently designing and analysing the clinical trials of tomorrow. PMID- 11858988 TI - Addressing the challenge of intergroup studies in oncology: the EORTC experience. European Organisation for Research and Treatment of Cancer. AB - Intergroup studies are conducted by more than one clinical research group. There are several difficulties that hamper in practice the possibility of conducting such trials, as all interested parties will have to address unusual and complex issues. These are mainly related to differences in size, interests, motivations and means among different research organisations. The EORTC recognises the importance to promote intergroup collaboration providing to all interested groups the necessary expertise and organisational support to conduct intergroup studies. The role of the EORTC evolved from the spontaneous organisations of intergroup trials to the definition of a basic set of principles and criteria that groups have to fulfil to participate in intergroup trials. Recently, a specific EORTC Intergroup Office started its activity devoted to solve the issues related to the intergroup co-operation. This office will have an increasing role to promote and help in conducting intergroup studies. PMID- 11858989 TI - New 2-(1-adamantylcarbonyl)pyridine and 1-acetyladamantane thiosemicarbazones thiocarbonohydrazones: cell growth inhibitory, antiviral and antimicrobial activity evaluation. AB - The new thiosemicarbazones and thiocarbonohydrazones derived from 2-(1 adamantylcarbonyl)pyridine and 1-acetyladamantane were synthesized and evaluated for their inhibitory effect on tumor cell proliferation and their antiviral and antimicrobial activity. Thiosemicarbazone inhibited tumor cell proliferation (GI50's range: 2.4-100 microM and mean GI50 43.9 microM against various human leukemic cell lines) while thiosemicarbazone and thiocarbonohydrazone 5d exhibited significant inhibition of tumor cell proliferation (GI50's range 2.3 23.6 microM and mean GI50 7.2 microM for and GI50's range 2.4-32.4 microM and mean GI50 12.8microM for ). These GI50 values are comparable to that of 2 acetylpyridine thiosemicarbazone an important lead in TSC's family. The compounds did not afford specific activity against any of the viruses tested when examined at non-toxic concentrations. A weak activity was found for thiocarbonohydrazones against Gram-(+) bacteria (MIC(50) 117.3 and 133 microM, respectively). Using a combination of molecular mechanics calculations and NOE spectroscopy it was shown that the parent compounds and have opposite configuration around C=N bond. Whether this difference in structure can be correlated with the biological activity will be investigated in future studies. PMID- 11858990 TI - N-aryl 2,6-dimethoxybiphenylalanine analogues as VLA-4 antagonists. AB - A series of N-arylated phenylalanine derivatives has been synthesized and has been shown to be potent inhibitors of the integrin VLA-4. N-phenyl and N heteroaryl derivatives with hydrogen bond acceptors in the meta position demonstrated low nanomolar activity against VLA-4. PMID- 11858991 TI - Structure-based design, synthesis, and biological evaluation of irreversible human rhinovirus 3C protease inhibitors. Part 7: structure-activity studies of bicyclic 2-pyridone-containing peptidomimetics. AB - The structure-based design, chemical synthesis, and biological evaluation of bicyclic 2-pyridone-containing human rhinovirus (HRV) 3C protease (3CP) inhibitors are described. An optimized compound is shown to exhibit antiviral activity when tested against a variety of HRV serotypes (EC(50)'s ranging from 0.037 to 0.162 microM). PMID- 11858992 TI - Anti-HIV-1 activities and pharmacokinetics of new arylpiperazinyl fluoroquinolones. AB - Anti-HIV-1 activities and pharmacokinetics of a series of novel arylpiperazinyl fluoroquinolones are reported. Modification at the C-8 position with a trifluoromethyl group was superior to that with a difluoromethoxy group to achieve higher anti-HIV-1 activity. Two compounds studied exhibited quite high anti-HIV-1 activities (IC(50)<50 nM) in vitro and high bioavailabilities (BA>90%) in monkeys. PMID- 11858994 TI - Synthesis of an alpha-fucosidase inhibitor, 5a-carba-beta-L-fucopyranosylamine, and fucose-type alpha- and beta-DL-valienamine unsaturated derivatives. AB - Discovery of a very potent alpha-fucosidase inhibitor 5a-carba-alpha-L fucopyranosylamine led to preparation of its beta-anomer and the respective unsaturated derivatives, fucose-type alpha- and beta-valienamines, in order to elucidate the structure-activity relationship of carba-aminosugar inhibitors of this kind. Compound was demonstrated to be a potent inhibitor (K(i)=2.0 x 10(-7) M, bovine kidney), possessing ca. one-tenth of the activity of the parent. Interestingly, and were found to be rather weak inhibitors, contrary to the expectations based on the activity relationships between the alpha-glucosidase inhibitors, alpha-glucose-type validamine and valienamine. PMID- 11858993 TI - Novel, potent non-covalent thrombin inhibitors incorporating p(3)-lactam scaffolds. AB - Evolution of P(1)-argininal inhibitor prototypes led to a series of non-covalent P(3)-7-membered lactam inhibitors 1a-w, featuring novel peptidomimetic units that probe each of the S(1), S(2), and S(3) specificity pockets of thrombin. Rigid P(1)-arginine surrogates possessing a wide range of basicity (calcd pK(a)'s approximately neutral-14) were surveyed. The design, synthesis, and biological activity of these targets are presented. PMID- 11858995 TI - Anionic cyclophanes as potential reversal agents of muscle relaxants by chemical chelation. AB - A series of carboxyl-containing cyclophanes have been designed and synthesised as chemical chelators (or host molecules) of cationic muscle relaxant drugs (or guest molecules). Three of these cyclophane derivatives, 1-3, have been shown by NMR to form 1:1 complexes with the muscle relaxants pancuronium, and gallamine, in D(2)O, with association constants up to 10(4) M(-1). When tested in an in vitro chick biventer muscle preparation, the cyclophanes reversed the neuromuscular block induced by pancuronium and gallamine, with having the most effective reversal against pancuronium (EC(50) 40 microM. PMID- 11858996 TI - Novel potent antagonists of human neuropeptide Y Y5 receptors. Part 2: substituted benzo[a]cycloheptene derivatives. AB - Novel benzo[a]cycloheptene derivatives were prepared for the purpose of searching new neuropeptide Y-Y5 (NPY-Y5) receptor antagonists. The structure-activity relationships are described and compound 2o (FR226928) showed the most potent affinity for Y5 receptor of all we prepared and was found to have higher potency and better selectivity for Y5 over Y1 receptor affinities when compared with the known lead compound 1. PMID- 11858997 TI - Peptidomimetics of efflux pump inhibitors potentiate the activity of levofloxacin in Pseudomonas aeruginosa. AB - Several classes of peptidomimetics of the efflux pump inhibitor D-ornithine-D homophenylalanine-3-aminoquinoline (MC-02,595) have been prepared and evaluated for their ability to potentiate the activity of the fluoroquinolone levofloxacin in Pseudomonas aeruginosa. A number of the new analogues were as active or more active than the lead, demonstrating that a peptide backbone is not essential for activity. PMID- 11858998 TI - Benzamide-based thiolcarbamates: a new class of HIV-1 NCp7 inhibitors. AB - The HIV-1 nucleocapsid protein NCp7, which contains two highly conserved zinc fingers, is being used as a novel target for AIDS therapy due to its pivotal role in viral replication and its mutationally intolerant nature. Herein we report a new class of NCp7 inhibitors that possess good antiviral activity with low cellular toxicity. PMID- 11859000 TI - The synthesis and selective IL-2 inhibitory activity of bis piperazine-phenol Mannich adducts. AB - Novel phenol bis-Mannich adducts were identified as IL-2 expression inhibitors in a T cell proliferation screening assay. Analogues of the lead compound were prepared through parallel synthesis and a highly selective IL-2 inhibitor was discovered that provided a suitable compound for further optimization. PMID- 11858999 TI - Novel agents combining platelet activating factor (PAF) receptor antagonist with thromboxane synthase inhibitor (TxSI). AB - Compounds 1 or 2 which possess dual-acting PAF antagonist/TxSI in a previous paper were modified and evaluated for the dual-acting activity. It was found that several compounds were potent dual-acting PAF antagonist/TxSI in and ex vivo. 6 (2-Chlorophenyl)-3-[4-[(E/Z)-6-ethoxycarbonyl-1-(3-pyridyl)-1 hexenyl]phenylmethyl]-8,11-dimethyl-2,3,4,5-tetrahydro-8H-pyrido[4',3': 4,5]thieno[3,2-f]triazolo[4,3-a]diazepine (12) is excellent orally dual-acting PAF antagonist/TxSI. PMID- 11859001 TI - Synthesis and activity of a new methoxytetrahydropyran derivative as dual cyclooxygenase-2/5-lipoxygenase inhibitor. AB - Dual COX-2/5-LO inhibitors are described as potential new therapeutic agents for inflammatory diseases. A surprisingly potent effect of a 5-LO pharmacophoric group on the COX-2 inhibition is presented as well as pharmacological in vitro and in vivo results. PMID- 11859003 TI - Potent nonsteroidal progesterone receptor agonists: synthesis and SAR study of 6 aryl benzoxazines. AB - Novel 6-aryl benzoxazines were prepared and examined as progesterone receptor (PR) modulators. In contrast to the structurally related 6-aryl dihydroquinoline PR antagonists, the 6-aryl benzoxazines were potent PR agonists. Compounds 4e, 5b, and 6a with the 2,4,4-trimethyl-1,4-dihydro-2H-benzo[d][1,3]oxazine core were the most potent PR agonists in the series with sub-nanomolar activities (EC(50) 0.20-0.35nM). Compound 6a was more potent than progesterone (P4) in the in vivo decidualization assay in an ovariectomized female rat model by subcutaneous administration with an ED(50) of 1.5mg/kg (vs 5.62mg/kg for P4). PMID- 11859002 TI - Synthesis and vasorelaxant activity of new coumarin and furocoumarin derivatives. AB - We have synthesized a new series of coumarins and furocoumarins and evaluated their vasorelaxant activity in rat aorta rings pre-contracted with noradrenaline or by depolarisation with high KCl. The new furocoumarins relax smooth vascular muscle with a profile similar to that of khellin (a furochromone that directly relaxes smooth muscle) and at a greater potency, suggesting that these compounds have a potential interest for the development of new and more efficient vasodilator drugs. PMID- 11859004 TI - Substituted 2-(R)-methyl piperazines as muscarinic M(2) selective ligands. AB - A novel series of 2-(R)-methyl-substituted piperazines (e.g., 2) is described. They are potent M(2) selective ligands that have >100-fold selectivity versus the M(1) receptor. In the rat microdialysis assay, compound 14 showed significantly enchanced levels of acetylcholine after oral administration. PMID- 11859005 TI - Synthesis and structure-activity relationships of M(2)-selective muscarinic receptor ligands in the 1-[4-(4-arylsulfonyl)-phenylmethyl]-4-(4-piperidinyl) piperazine family. AB - The synthesis and muscarinic binding properties of compounds based on the 1-[4-(4 arylsulfonyl)phenylmethyl]-4-(1-aroyl-4-piperidinyl)-piperazine skeleton are described. For compounds, substituted with appropriately configured methyl groups at the benzylic center and at the piperazine 2-position, high levels of selective, M(2) subtype affinity could be obtained, particularly when the terminal N-aroyl residue was ortho-substituted. PMID- 11859006 TI - Novel potent antagonists of human neuropeptide Y Y5 receptors. Part 3: 7-methoxy 1-hydroxy-1-substituted tetraline derivatives. AB - As a part of our continuing research on NPY-Y5 receptor antagonists in the series of novel 6-methoxybenzo[a]cycloheptene derivatives, we discovered a novel skeleton, 7-methoxy-1-hydroxytetraline 7 which had been used as an intermediate, to be more suitable for increasing potencies leading to compound 3 (FR230481). Additionally, we discovered that the naphthalenesulfonamide moiety which was thought to be an essential pharmacophore could be replaced by the 5 chlorobenzothiazolin-3-acetic acid moiety to lead to potent compound 4 (FR233118). The structure-activity relationships on compounds 3,4 and their related derivatives are described. Unfortunately, although compounds 3 and 4 had very high affinities for Y5 receptors, their poor permeabilities to brain were shown by exo-vivo binding assays when orally administered. PMID- 11859007 TI - Synthesis of Sordaricin analogues as potent antifungal agents against Candida albicans. AB - Sordaricin derivatives possessing a cyclohexane ring appendage attached via an ether, thioether, amine, oxime, ester or amide linkage were synthesized and their antifungal activity was evaluated in vitro. Compounds containing a thioether bond or an oxime bond as a linkage exhibited potent MICs (< or = 0.125 microg/mL) against four Candida albicans strains including azole-low-susceptible strains. They were also active (MIC < or = 0.125 microg/mL) against Candida glabrata. Their in vivo efficacy was confirmed in a murine intravenous infection model with Candida albicans. PMID- 11859008 TI - Assamicin I and II, novel triterpenoid saponins with insulin-like activity from Aesculus assamica Griff. AB - Two novel triterpenoid saponins with insulin-like activity, termed assamicin I and II, were isolated from the roots of Aesculus assamica Griff. and their structures were characterized as 1 and 2, respectively. They inhibited release of free fatty acids from epinephrine-treated rat adipocytes and enhanced glucose uptake into 3T3-L1 adipocytes. PMID- 11859009 TI - Syntheses and binding affinities of 6-nitroquipazine analogues for serotonin transporter. Part 2: 4-substituted 6-nitroquipazines. AB - Eleven 4-substituted derivatives of 6-nitroquipazine were synthesized and evaluated for their abilities to displace [3H]citalopram binding to the rat cortical synaptic membranes. Among them, 4-chloro-6-nitroquipazine was shown to possess the highest binding affinity (K(i=)0.03 nM) which was approximately 6 times higher than that of 6-nitroquipazine (K(i)=0.17 nM) itself. In this paper, we describe the syntheses of 4-substituted 6-nitroquipazine derivatives, the results of corresponding biological evaluation and the SAR study. PMID- 11859010 TI - Synthesis and antiplatelet activity of gemfibrozil chiral analogues. AB - The chiral analogues of gemfibrozil 5-(2,5-dimethylphenoxy)-2-methylpentanoic acid and 5-(2,5-dimethylphenoxy)-2-ethylpentanoic acid were synthesized in optically active form using (S)-4-(1-methylethyl)-2-oxazolidinone as chiral auxiliary. All compounds inhibit human platelet aggregation. From these data, one can surmise that all tested compounds and gemfibrozil act at the platelet level with different mechanism than that of ASA, even if with a different potency. PMID- 11859011 TI - Cytotoxic, antifouling bromotyramines: a synthetic study on simple marine natural products and Their analogues. AB - Synthesis and biological evaluation of two naturally-occurring bromotyramines, moloka'iamine 1 and 3,5-dibromo-4-methoxy-beta-phenethylamine 2, together with several analogues, have been completed. Bromotyramine 2 is cytotoxic, and was found to be a potent antifoulant. Analogues 15 and 16 also displayed significant cytotoxic and antifouling activities. PMID- 11859012 TI - 2-Arylindoles as gonadotropin releasing hormone (GnRH) antagonists: optimization of the tryptamine side chain. AB - A series of 2-arylindoles containing novel heteroaromatic substituents on the tryptamine tether, based on compound 1, was prepared and evaluated for their ability to act as gonadotropin releasing hormone (GnRH) antagonists. Successful modifications of 1 included chain length variation (reduction) and replacement of the pyridine with heteroaromatic groups. These alterations culminated in the discovery of compound 27kk which had excellent in vitro potency and oral efficacy in rodents. PMID- 11859015 TI - The Golgi apparatus: going round in circles? AB - Recent studies have questioned the idea that the Golgi complex is a stable organelle with a unique identity through which secretory cargo is transported by vesicles. Instead, it is proposed that Golgi apparatus proteins continuously recycle via the endoplasmic reticulum by vesicle transport, whereas cargo molecules remain in maturing cisternal structures. Rather than forming a rigid matrix, structural Golgi proteins might be highly dynamic and recycle via the cytoplasm. I will discuss the evidence for these claims and consider whether or not they really disprove older ideas on how the Golgi apparatus is structured and performs its function. PMID- 11859013 TI - Structure-activity relationships of oxime neurokinin antagonists: oxime modifications. AB - A thorough SAR study of the oxime region of the dual NK(1)/NK(2) antagonist 1 revealed several modifications that result in potent dual antagonists. Follow up SAR studies on a second-generation scaffold demonstrate that certain polar groups on the oxime can improve the dual binding affinity to the subnanomolar range. PMID- 11859016 TI - A phosphorylation-driven ubiquitination switch for cell-cycle control. AB - Cellular changes in state can be dictated by complex all-or-nothing switches built from ultrasensitive protein kinase cascades, positive-feedback loops and other mechanisms. Recent work has established that phosphorylation-driven protein destruction through the SCF ubiquitin-ligase pathway can also occur in a switch like manner. In this context, multiple phosphorylation events are used to set a threshold for substrate targeting, thereby providing a framework for understanding the inter-relationship between protein phosphorylation and ubiquitin-mediated proteolysis. PMID- 11859022 TI - Crick's DNA papers available for all. PMID- 11859023 TI - The lamellipodium: where motility begins. AB - Lamellipodia, filopodia and membrane ruffles are essential for cell motility, the organization of membrane domains, phagocytosis and the development of substrate adhesions. Their formation relies on the regulated recruitment of molecular scaffolds to their tips (to harness and localize actin polymerization), coupled to the coordinated organization of actin filaments into lamella networks and bundled arrays. Their turnover requires further molecular complexes for the disassembly and recycling of lamellipodium components. Here, we give a spatial inventory of the many molecular players in this dynamic domain of the actin cytoskeleton in order to highlight the open questions and the challenges ahead. PMID- 11859024 TI - Partners-in-infection: host proteins involved in the transformation of plant cells by Agrobacterium. AB - Genetic modification of plant cells by Agrobacterium is the only known natural example of DNA transport between kingdoms. While the bacterial factors involved in Agrobacterium infection have been relatively well characterized, studies of their host cellular partners are just beginning. Here, we describe the plant cell factors that might participate in Agrobacterium-mediated genetic transformation and discuss their possible roles in this process. Because Agrobacterium probably adapts existing cellular processes for its life cycle, identifying the host factors participating in Agrobacterium infection might contribute to a better understanding of such basic biological processes as cell communication, intracellular transport and DNA repair and recombination as well as help expand the host range of Agrobacterium as a genetic engineering tool. PMID- 11859025 TI - Arresting developments in heptahelical receptor signaling and regulation. AB - It is well established that the function of most heptahelical receptors (seven transmembrane-span receptors; 7TMRs) is tightly regulated by the desensitizing actions of arrestins. Desensitization is the waning of 7TMR-mediated signals after prolonged exposure to agonist and occurs when arrestins bind to agonist occupied and phosphorylated receptors, uncoupling the receptors from G proteins and preventing further signaling. Recently, there has been a marked shift in the focus of research into arrestin function because it has become clear that they not only prevent signaling from 7TMRs but also initiate and direct new signals from the very 7TMRs that they desensitize. PMID- 11859026 TI - Signaling by protein phosphatases in the nucleus. AB - The nucleus contains a large variety of protein phosphatases, which function in key processes such as cell-cycle progression, replication, transcription and RNA processing. Here, we review the pleiotropic action of nuclear protein phosphatases and focus in particular on the underlying signaling strategies. It appears that nuclear protein phosphatases can both mediate and antagonize signaling by protein kinases, sometimes as part of feedback loops. Some protein phosphatases shuttle between the cytoplasm and the nucleus, which enables them to act as signal transducers between both compartments. An emerging theme is the contribution of protein phosphatases to cycles of protein phosphorylation and dephosphorylation that steer the assembly and firing of molecular machines in the nucleus. PMID- 11859027 TI - Functional diversity of protein C-termini: more than zipcoding? AB - The carboxylated (C)-terminus of proteins, which includes the single terminal alpha-carboxyl group and preceding residues, is uniquely positioned to serve as a recognition signature for a variety of cell-biological processes, including protein targeting, subcellular anchoring and the static and dynamic formation of macromolecular complexes. The terminal sequence motifs can be processed by posttranslational modifications, thereby providing a means to increase sequence diversity and to regulate interactions. Several classes of protein domains have been identified that are either designed for or are capable of interacting with protein C-termini - these include PDZ and TPR domains. The interactions between these protein domains and various terminal epitopes play an important role in specifying cell-biological functions. The combination of diversity and the plasticity of the chemistry of C-termini provides mechanisms for spatial and temporal specificity that are exploited by a variety of biological processes, ranging from specifying prokaryotic protein degradation to nucleating mammalian neuronal signaling complexes. Understanding the diverse functions of protein C termini might also provide an important indexing criterion for functional proteomics. PMID- 11859028 TI - Susan L. Lindquist. PMID- 11859030 TI - Recombinant human parathyroid hormone. PMID- 11859031 TI - Guidelines for managing community acquired pneumonia in adults. PMID- 11859032 TI - Academic medicine: a faltering engine. PMID- 11859033 TI - Yellow fever vaccine. PMID- 11859034 TI - Doctors working in prisons. PMID- 11859037 TI - Smoking increases among teenagers in eastern Europe. PMID- 11859038 TI - Monitoring body criticises hospital for mixed sex wards. PMID- 11859040 TI - DNA needs to be protected by law. PMID- 11859041 TI - Proposed genetic database on Tongans opposed. PMID- 11859042 TI - Doctors' chambers--pipe dream or blueprint for the future? PMID- 11859043 TI - Survey shows serious shortage of medical academics in the UK. PMID- 11859044 TI - Low consumption of seafood in early pregnancy as a risk factor for preterm delivery: prospective cohort study. AB - OBJECTIVE: To determine the relation between intake of seafood in pregnancy and risk of preterm delivery and low birth weight. DESIGN: Prospective cohort study. SETTING: Aarhus, Denmark. PARTICIPANTS: 8729 pregnant women. MAIN OUTCOME MEASURES: Preterm delivery and low birth weight. RESULTS: The occurrence of preterm delivery differed significantly across four groups of seafood intake, falling progressively from 7.1% in the group never consuming fish to 1.9% in the group consuming fish as a hot meal and an open sandwich with fish at least once a week. Adjusted odds for preterm delivery were increased by a factor of 3.6 (95% confidence interval 1.2 to 11.2) in the zero consumption group compared with the highest consumption group. Analyses based on quantified intakes indicated that the working range of the dose-response relation is mainly from zero intake up to a daily intake of 15 g fish or 0.15 g n-3 fatty acids. Estimates of risk for low birth weight were similar to those for preterm delivery. CONCLUSIONS: Low consumption of fish was a strong risk factor for preterm delivery and low birth weight. In women with zero or low intake of fish, small amounts of n-3 fatty acids--provided as fish or fish oil--may confer protection against preterm delivery and low birth weight. PMID- 11859045 TI - Clinical course of hepatitis C virus during the first decade of infection: cohort study. AB - OBJECTIVE: To determine the clinical course of hepatitis C virus in the first decade of infection in a group of patients who acquired their infections on a known date. DESIGN: Cohort study. SETTING: Clinical centres throughout the United Kingdom. PARTICIPANTS: 924 transfusion recipients infected with the hepatitis C virus (HCV) traced during the HCV lookback programme and 475 transfusion recipients who tested negative for antibodies to HCV (controls). MAIN OUTCOME MEASURES: Clinical evidence of liver disease and survival after 10 years of infection. RESULTS: All cause mortality was not significantly different between patients and controls (Cox's hazards ratio 1.41, 95% confidence interval 0.95 to 2.08). Patients were more likely to be certified with a death related to liver disease than were controls (12.84, 1.73 to 95.44), but although the risk of death directly from liver disease was higher in patients than controls this difference was not significant (5.78, 0.72 to 46.70). Forty per cent of the patients who died directly from liver disease were known to have consumed excess alcohol. Clinical follow up of 826 patients showed that liver function was abnormal in 307 (37.2%), and 115 (13.9%) reported physical signs or symptoms of liver disease. Factors associated with developing liver disease were testing positive for HCV ribonucleic acid (odds ratio 6.44, 2.67 to 15.48), having acquired infection when older (at age > or = 40 years; 1.80, 1.14 to 2.85), and years since transfusion (odds ratio 1.096 per year, 1.00 to 1.20). For patients with severe disease, sex was also significant (odds ratio for women 0.38, 0.17 to 0.88). Of the 362 patients who had undergone liver biopsy, 328 (91%) had abnormal histological results and 35 (10%) of these were cirrhotic. CONCLUSIONS: Hepatitis C virus infection did not have a great impact on all cause mortality in the first decade of infection. Infected patients were at increased risk of dying directly from liver disease, particularly if they consumed excess alcohol, but this difference was not statistically significant. PMID- 11859046 TI - Ultrasonography and abdominal radiography versus intravenous urography in investigation of urinary tract infection in men: prospective incident cohort study. AB - OBJECTIVES: To compare ultrasonography and abdominal radiography with intravenous urography in the investigation of urinary tract infection in men. DESIGN: Prospective study in two hospital departments. Radiological procedures and urological assessments performed on different days by different clinicians SETTING: District general hospital. PARTICIPANTS: Consecutive series of men (n=114) referred to the department of urology for investigation of proved urinary tract infection. INTERVENTIONS: Ultrasonography and intravenous urography of renal tract and assessment of urinary flow rate. Clinical assessment, cystoscopy, urodynamic studies, and transrectal ultrasonography with biopsy. MAIN OUTCOME MEASURES: Sensitivity and specificity of ultrasonography and abdominal radiography compared with intravenous urography. RESULTS: Important abnormalities were seen in 53 of 100 fully evaluated patients, the most common being a poorly emptying bladder (34). The combination of plain radiographs of kidneys, ureter, and bladder and ultrasonography detected more abnormalities than intravenous urography alone. No important abnormality was missed by this combination (sensitivity 100% and specificity 93%). CONCLUSIONS: Ultrasonography with abdominal radiography is as accurate as intravenous urography in detecting important urological abnormalities in men presenting with urinary tract infection. This combination is safer than intravenous urography and should be the initial investigation for such patients. Additional determination of urinary flow rate is useful for the assessment of an incompletely emptying bladder. PMID- 11859047 TI - Association between childhood fractures and poverty: population based study. PMID- 11859048 TI - Denial of pregnancy: population based study. PMID- 11859050 TI - Experiences and career intentions of general practice registrars in Thames deaneries: postal survey. PMID- 11859049 TI - Comparison of estimates and calculations of risk of coronary heart disease by doctors and nurses using different calculation tools in general practice: cross sectional study. AB - OBJECTIVE: To assess the effect of using different risk calculation tools on how general practitioners and practice nurses evaluate the risk of coronary heart disease with clinical data routinely available in patients' records. DESIGN: Subjective estimates of the risk of coronary heart disease and results of four different methods of calculation of risk were compared with each other and a reference standard that had been calculated with the Framingham equation; calculations were based on a sample of patients' records, randomly selected from groups at risk of coronary heart disease. SETTING: General practices in central England. PARTICIPANTS: 18 general practitioners and 18 practice nurses. MAIN OUTCOME MEASURES: Agreement of results of risk estimation and risk calculation with reference calculation; agreement of general practitioners with practice nurses; sensitivity and specificity of the different methods of risk calculation to detect patients at high or low risk of coronary heart disease. RESULTS: Only a minority of patients' records contained all of the risk factors required for the formal calculation of the risk of coronary heart disease (concentrations of high density lipoprotein (HDL) cholesterol were present in only 21%). Agreement of risk calculations with the reference standard was moderate (kappa=0.33-0.65 for practice nurses and 0.33 to 0.65 for general practitioners, depending on calculation tool), showing a trend for underestimation of risk. Moderate agreement was seen between the risks calculated by general practitioners and practice nurses for the same patients (kappa=0.47 to 0.58). The British charts gave the most sensitive results for risk of coronary heart disease (practice nurses 79%, general practitioners 80%), and it also gave the most specific results for practice nurses (100%), whereas the Sheffield table was the most specific method for general practitioners (89%). CONCLUSIONS: Routine calculation of the risk of coronary heart disease in primary care is hampered by poor availability of data on risk factors. General practitioners and practice nurses are able to evaluate the risk of coronary heart disease with only moderate accuracy. Data about risk factors need to be collected systematically, to allow the use of the most appropriate calculation tools. PMID- 11859051 TI - Peripheral neuropathy. PMID- 11859052 TI - Introduction. II--basic terminology. PMID- 11859053 TI - Does animal experimentation inform human healthcare? Observations from a systematic review of international animal experiments on fluid resuscitation. PMID- 11859054 TI - Evaluation of diagnostic procedures. PMID- 11859055 TI - Postoperative starvation after gastrointestinal surgery. Type of intravenous nutrition given in control groups is not indicated. PMID- 11859056 TI - Cardiotocography v Doppler auscultation. Guidelines highlight gaps in research evidence. PMID- 11859057 TI - Gastro-oesophageal reflux disease in general practice. Last year's guidelines are probably biased. PMID- 11859058 TI - Caesarean section rate in England and Wales. Maybe midwives were too busy. PMID- 11859059 TI - Self management is the future. PMID- 11859060 TI - Management of chronic disease. System needs managing better. PMID- 11859067 TI - p53 binds telomeric single strand overhangs and t-loop junctions in vitro. AB - The interaction of p53 with a human model telomere in vitro was examined by electron microscopy. p53 demonstrated a sequence-independent affinity for telomeric DNA in vitro, localizing to the 3' single strand overhang and the t loop junction both in the presence and absence of associated TRF2. Binding was not observed above background along the duplex telomeric repeats. However, the efficiency of TRF2-catalyzed t-loop formation on the model DNA was increased 2 fold in the presence of p53 although a variety of single strand or Holliday junction-binding proteins did not facilitate t-loop formation. These results suggest that p53 has an active role in telomere maintenance and structure through association with the t-loop junction. PMID- 11859068 TI - Histone acetylation status and DNA sequence modulate ATP-dependent nucleosome repositioning. AB - A cell-free system derived from Drosophila embryos was used to investigate positioning of nucleosomes on specific DNA sequences. This system can be used to reconstitute differently acetylated nucleosome arrays possessing ATP-dependent dynamic properties that are not observed with chromatin assembled from pure components. Nucleosome positioning on different DNA sequences was studied by restriction endonuclease assay. The sequence of DNA and the acetylation status of histones had profound effects on the distribution of nucleosomes, suggesting their cooperative effect on nucleosome repositioning. PMID- 11859069 TI - Plasmodium falciparum histidine-rich protein-2 (PfHRP2) modulates the redox activity of ferri-protoporphyrin IX (FePPIX): peroxidase-like activity of the PfHRP2-FePPIX complex. AB - Histidine-rich protein-2 from Plasmodium falciparum (PfHRP2) binds up to 50 molecules of ferri-protoporphyrin IX (FePPIX) (Choi, C. Y., Cerda, J. F., Chu, H. A., Babcock, G. T., and Marletta, M. A. (1999) Biochemistry 38, 16916-16924). We reasoned that the PfHRP2-FePPIX complex has antioxidant properties that could be beneficial to the parasite. Therefore, we examined whether binding to PfHRP2 modulated the redox properties of FePPIX. We observed that PfHRP2 completely inhibited the auto-oxidation of ascorbate mediated by free FePPIX. We also investigated the peroxidase activity of PfHRP2-FePPIX using 13-hydroperoxy-9,11 octadienoate (18:2-OOH) as substrate. Reaction of PfHRP2-FePPIX with 18:2-OOH in the presence of added reducing agents gave 13-hydroxy-9,11-octadienoate (18:2-OH) as a major product and 13-keto-9,11-octadienoate (18:2=O) and 9,12,13-trihydroxy 10-octadecaenoate as minor products. Binding of FePPIX to PfHRP2 lowered the rate of decomposition of 18:2-OOH and increased the 18:2-OH to 18:2=O ratio. Similar to other authentic peroxidases, phenols, amines, and biological reductants like ascorbate promoted 18:2-OH production, and NaCN inhibited 18:2-OH production. Thioanisole also acted as a reductant and was converted to thioanisole sulfoxide, suggesting formation of compound I during the reaction. These data show that PfHRP2 modulates the redox activity of FePPIX and that the PfHRP2-FePPIX complex may have previously unrecognized antioxidant properties. PMID- 11859070 TI - Characteristics of brain Kv1 channels tailored to mimic native counterparts by tandem linkage of alpha subunits: implications for K+ channelopathies. AB - Most neuronal Kv1 channels contain Kv1.1, Kv1.2 alpha, and Kvbeta2.1 subunits, yet the influences of their stoichiometries on properties of the (alpha)(4)(beta)(4) variants remain undefined. cDNAs were engineered to contain 0, 1, 2, or 4 copies of Kv1.1 with the requisite number of Kv1.2 and co-expressed in mammalian cells with Kvbeta2.1 to achieve "native-like" hetero-oligomers. The monomeric (Kv1.1 or 1.2), dimeric (Kv1.1-1.2 or 1.2-1.2), and tetrameric (Kv1.1 (1.2)(3)) constructs produced proteins of M(r) approximately 62,000, 120,000, and 240,000, which assembled into (alpha)(4)(beta)(4) complexes. Each alpha cRNA yielded a distinct K(+) current in oocytes, with voltage dependence of activation being shifted negatively as the Kv1.1 content in tetramers was increased. Channels containing 1, 2, or 4 copies of Kv1.1 were blocked by dendrotoxin k (DTX)(k) with similarly high potencies, whereas Kv(1.2)(4) proved nonsusceptible. Accordingly, Kv1.2/beta2.1 expressed in baby hamster kidney cells failed to bind DTX(k); in contrast, oligomers containing only one Kv1.1 subunit in a tetramer exhibited high affinity, with additional copies causing modest increases. Thus, one Kv1.1 subunit largely confers high affinity for DTX(k), whereas channel electrophysiological properties are tailored by the content of Kv1.1 relative to Kv1.2. This notable advance could explain the diversity of symptoms of human episodic ataxia I, which is often accompanied by myokymia, due to mutated Kv1.1 being assembled in different combinations with wild-type and Kv1.2. PMID- 11859071 TI - Uncoupling of the cholera toxin-G(M1) ganglioside receptor complex from endocytosis, retrograde Golgi trafficking, and downstream signal transduction by depletion of membrane cholesterol. AB - To induce toxicity, cholera toxin (CT) must first bind ganglioside G(M1) at the plasma membrane, enter the cell by endocytosis, and then traffic retrograde into the endoplasmic reticulum. We recently proposed that G(M1) provides the sorting motif necessary for retrograde trafficking into the biosynthetic/secretory pathway of host cells, and that such trafficking depends on association with lipid rafts and lipid raft function. To test this idea, we examined whether CT action in human intestinal T84 cells depends on membrane cholesterol. Chelation of cholesterol with 2-hydroxypropyl beta-cyclodextrin or methyl beta-cyclodextrin reversibly inhibited CT-induced chloride secretion and prolonged the time required for CT to enter the cell and induce toxicity. These effects were specific to CT, as identical conditions did not alter the potency or toxicity of anthrax edema toxin that enters the cell by another mechanism. We found that endocytosis and trafficking of CT into the Golgi apparatus depended on membrane cholesterol. Cholesterol depletion also changed the density and specific protein content of CT-associated lipid raft fractions but did not entirely displace the CT-G(M1) complex from these lipid raft microdomains. Taken together these data imply that cholesterol may function to couple the CT-G(M1) complex with raft domains and with other membrane components of the lipid raft required for CT entry into the cell. PMID- 11859072 TI - Autocrine-mediated activation of STAT3 correlates with cell proliferation in breast carcinoma lines. AB - The intracellular signals driving the proliferation of breast carcinoma (BC) cells have been widely studied. Both the mitotic and metastatic potential of BC cells have been linked to the frequent overexpression of ErbB family members. Other signaling molecules, including the estrogen receptor, the tyrosine kinases c-Src and Syk, and STAT proteins, especially STAT3, have also been implicated in BC tumor growth. Here we have examined ErbB and STAT protein expression and activation in six BC-derived cell lines. ErbB expression and tyrosine phosphorylation varied considerably among the six cell lines. However, STAT protein expression and activation were more consistent. Two levels of STAT3 activation were distinguished in DNA-binding assays: an epidermal growth factor inducible, high level that requires both ErbB1 and Janus kinase (JAK) activity and an elevated serum-dependent level that is maintained by autocrine/paracrine signaling and requires JAK activity but is independent of ErbB1 kinase activity. BC cell growth could be inhibited by dominant-negative versions of STAT3 and the JAK inhibitor AG490 but not by PD153035 or PD168393, inhibitors of ErbB1 kinase activity. This indicates that BC cell proliferation may be a consequence of STAT3 activation by autocrine/paracrine signals. PMID- 11859073 TI - Analysis of a multicomponent thermostable DNA polymerase III replicase from an extreme thermophile. AB - This report takes a proteomic/genomic approach to characterize the DNA polymerase III replication apparatus of the extreme thermophile, Aquifex aeolicus. Genes (dnaX, holA, and holB) encoding the subunits required for clamp loading activity (tau, delta, and delta') were identified. The dnaX gene produces only the full length product, tau, and therefore differs from Escherichia coli dnaX that produces two proteins (gamma and tau). Nonetheless, the A. aeolicus proteins form a taudeltadelta' complex. The dnaN gene encoding the beta clamp was identified, and the taudeltadelta' complex is active in loading beta onto DNA. A. aeolicus contains one dnaE homologue, encoding the alpha subunit of DNA polymerase III. Like E. coli, A. aeolicus alpha and tau interact, although the interaction is not as tight as the alpha-tau contact in E. coli. In addition, the A. aeolicus homologue to dnaQ, encoding the epsilon proofreading 3'-5'-exonuclease, interacts with alpha but does not form a stable alpha.epsilon complex, suggesting a need for a brace or bridging protein to tightly couple the polymerase and exonuclease in this system. Despite these differences to the E. coli system, the A. aeolicus proteins function to yield a robust replicase that retains significant activity at 90 degrees C. Similarities and differences between the A. aeolicus and E. coli pol III systems are discussed, as is application of thermostable pol III to biotechnology. PMID- 11859074 TI - Phosphatidylinositol 3-kinase/Akt signaling is neither required for hypoxic stabilization of HIF-1 alpha nor sufficient for HIF-1-dependent target gene transcription. AB - The serine/threonine kinase Akt/PKB and the oxygen-responsive transcription factor HIF-1 share the ability to induce such processes as angiogenesis, glucose uptake, and glycolysis. Akt activity and HIF-1 are both essential for development and implicated in tumor growth. Upon activation by products of phosphatidylinositol 3-kinase (PI3K), Akt phosphorylates downstream targets that stimulate growth and inhibit apoptosis. Previous reports suggest that Akt may achieve its effects on angiogenesis and glucose metabolism by stimulating HIF-1 activity. We report here that, whereas serum stimulation can induce a slight accumulation of HIF-1 alpha protein in a PI3K/Akt pathway-dependent fashion, hypoxia induces much higher levels of HIF-1 alpha protein and HIF-1 DNA binding activity independently of PI3K and mTOR activity. In addition, we find the effects of constitutively active Akt on HIF-1 activity are cell-type specific. High levels of Akt signaling can modestly increase HIF-1 alpha protein, but this increase does not affect HIF-1 target gene expression. Therefore, the PI3K/Akt pathway is not necessary for hypoxic induction of HIF-1 subunits or activity, and constitutively active Akt is not itself sufficient to induce HIF-1 activity. PMID- 11859075 TI - The cyclin-ubiquitin ligase activity of cyclosome/APC is jointly activated by protein kinases Cdk1-cyclin B and Plk. AB - The cyclosome/anaphase-promoting complex is a multisubunit ubiquitin ligase that targets for degradation mitotic cyclins and some other cell cycle regulators in exit from mitosis. It becomes enzymatically active at the end of mitosis. The activation of the cyclosome is initiated by its phosphorylation, a process necessary for its conversion to an active form by the ancillary protein Cdc20/Fizzy. Previous reports have implicated either cyclin-dependent kinase 1 cyclin B or polo-like kinase as the major protein kinase that directly phosphorylates and activates the cyclosome. These conflicting results could be due to the use of partially purified cyclosome preparations or of immunoprecipitated cyclosome, whose interactions with protein kinases or ancillary factors may be hampered by binding to immobilized antibody. To examine this problem, we have purified cyclosome from HeLa cells by a combination of affinity chromatography and ion exchange procedures. With the use of purified preparations, we found that both cyclin-dependent kinase 1-cyclin B and polo-like kinase directly phosphorylated the cyclosome, but the pattern of the phosphorylation of the different cyclosome subunits by the two protein kinases was not similar. Each protein kinase could restore only partially the cyclin ubiquitin ligase activity of dephosphorylated cyclosome. However, following phosphorylation by both protein kinases, an additive and nearly complete restoration of cyclin-ubiquitin ligase activity was observed. It is suggested that this joint activation may be due to the complementary phosphorylation of different cyclosome subunits by the two protein kinases. PMID- 11859076 TI - Vav1 couples T cell receptor to serum response factor-dependent transcription via a MEK-dependent pathway. AB - The Vav family of guanine nucleotide exchange factors for Rho family GTPases plays a critical role in lymphocyte proliferation, gene transcription, and cytoskeleton reorganization following immunoreceptor stimulation. However, its role in immediate early gene activation is unclear. In this study, we have investigated the mechanisms by which Vav1 can regulate c-fos serum response element transcriptional activity. We show that T cell antigen receptor (TCR) stimulation induces the phosphorylation of serum response factor (SRF) on serine 103 and increases the binding of SRF complexes on serum response element in a MEK and p38-dependent pathway. The physiological relevance of our findings is supported by the inhibition of the interleukin-2 gene transcriptional activity by a dominant negative SRF mutant. Overexpression of Vav1, which partially mimics TCR stimulation, promotes SRF-dependent transcription, and dominant negative Vav1 mutants block SRF activation by TCR. SRF activation by Vav1 occurs through a signaling cascade consisting of Rac1/Cdc42 and the serine/threonine kinases Pak1 and MEK, but independently of the phosphatidylinositol 3-kinase pathway. Interestingly, Vav2 also enhances SRF through Rho GTPases, suggesting that Vav proteins are general regulators of SRF activation in lymphocytes. This report establishes Vav proteins as a direct link between antigen receptors and SRF dependent early gene expression. PMID- 11859077 TI - Peroxisomal membrane protein import does not require Pex17p. AB - Of the approximately 20 proteins required for peroxisome biogenesis, only four have been implicated in the process of peroxisomal membrane protein (PMP) import: Pex3p, Pex16p, Pex17p, and Pex19p. To improve our understanding of the role that Pex17p plays in PMP import, we examined the behavior of PMPs in a Pichia pastoris pex17 mutant. Relative to wild-type cells, pex17 cells appeared to have a mild reduction in PMP stability and slightly aberrant PMP behavior in subcellular fractionation experiments. However, we also found that the behavior of PMPs in the pex17 mutant was indistinguishable from PMP behavior in a pex5 mutant, which has no defect in PMP import, and was far different from PMP behavior in a pex3 mutant, which has a bona fide defect in PMP import. Furthermore, we found that a pex14 mutant, which has no defect in PMP import, lacks detectable levels of Pex17p. Based on these and other results, we propose that Pex17p acts primarily in the matrix protein import pathway and does not play an important role in PMP import. PMID- 11859078 TI - Crystal structure of the N-terminal segment of human eukaryotic translation initiation factor 2alpha. AB - Eukaryotic translation initiation factor 2alpha (eIF2alpha) is a member of the eIF2 heterotrimeric complex that binds and delivers Met-tRNA(i)(Met) to the 40 S ribosomal subunit in a GTP-dependent manner. Phosphorylation/dephosphorylation of eIF2alpha at Ser-51 is the major regulator of protein synthesis in eukaryotic cells. Here, we report the first structural analysis on eIF2, the three dimensional structure of a 22-kDa N-terminal portion of human eIF2alpha by x-ray diffraction at 1.9 A resolution. This structure contains two major domains. The N terminus is a beta-barrel with five antiparallel beta-strands in an oligonucleotide binding domain (OB domain) fold. The phosphorylation site (Ser 51) is on the loop connecting beta3 and beta4 in the OB domain. A helical domain follows the OB domain, and the first helix has extensive interactions, including a disulfide bridge, to fix its orientation with respect to the OB domain. The two domains meet along a negatively charged groove with highly conserved residues, indicating a likely site for protein-protein interaction. PMID- 11859079 TI - Two defective heterozygous luteinizing hormone receptors can rescue hormone action. AB - Luteinizing hormone receptor is a G protein-coupled receptor and consists of two halves: the N-terminal extracellular half (exodomain) and C-terminal membrane associated half (endodomain). Hormone binds to the exodomain, and the resulting hormone-exodomain complex modulates the endodomain to generate signals. There are mutations that impair either hormone binding or signal generation. We report that the coexpression of a binding defective mutant and a signal-defective mutant rescues signal generation to produce cAMP. This rescue requires both types of mutant receptors and is dependent on the human chorionic gonadotropin dose, the surface concentration of mutant receptors, and the amino acid position of mutations. Furthermore, random collisions among mutant receptors are not involved in the rescue. Our observations provide new insights into the mechanisms of the functional and structural relationship of the exo- and endodomain, signal transduction, and receptor genetics, in particular for defective heterozygotes. PMID- 11859080 TI - The serotonin binding site of human and murine 5-HT2B receptors: molecular modeling and site-directed mutagenesis. AB - Bacteriorhodopsin and rhodopsin crystal structures were used as templates to build structural models of the mouse and human serotonin (5-HT)-2B receptors (5 HT(2B)Rs). Serotonin was docked to the receptors, and the amino acids predicted to participate to its binding were subjected to mutagenesis. 5-HT binding affinity and 5-HT-induced inositol triphosphate production were measured in LMTK( ) cells transfected with either wild-type or mutated receptor genes. According to these measurements, the bacteriorhodopsin-based models of the 5-HT(2B)Rs appear more confident than the rhodopsin-based ones. Residues belonging to the transmembrane domains 3 and 6, i.e. Asp(3.32), Ser(3.36), Phe(6.52), and Asn(6.55), make direct contacts with 5-HT. In addition, Trp(3.28), Phe(3.35), Phe(6.52), and Phe(7.38) form an aromatic box surrounding 5-HT. The specificity of human and mouse 5-HT(2B)Rs may be reflected by different rearrangements of the aromatic network upon 5-HT binding. Two amino acids close to Pro(5.50) in the human transmembrane domain 5 sequence were permuted to introduce a "mouse-like" sequence. This change was enough to confer the human 5-HT(2B)R properties similar to those of the mouse. Taken together, the computed models and the site-directed mutagenesis experiments give a structural explanation to (i) the different 5-HT pK(D) values measured with the human and mouse 5-HT(2B)Rs (7.9 and 5.8, respectively) and (ii) the specificity of 5-HT binding to 5-HT(2B)Rs as compared with other serotonergic G-protein coupled receptors. PMID- 11859082 TI - Probing the limits of electrostatic catalysis by uracil DNA glycosylase using transition state mimicry and mutagenesis. AB - The DNA repair enzyme uracil DNA glycosylase (UDG) hydrolyzes the glycosidic bond of deoxyuridine in DNA by a remarkable mechanism involving formation of a positively charged oxacarbenium ion-uracil anion intermediate. We have proposed that the positively charged intermediate is stabilized by being sandwiched between the combined negative charges of the anionic uracil leaving group and a conserved aspartate residue that are located on opposite faces of the sugar ring. Here we establish that a duplex DNA oligonucleotide containing a cationic 1-aza deoxyribose (I) oxacarbenium ion mimic is a potent inhibitor of UDG that binds tightly to the enzyme-uracil anion (EU(-)) product complex (K(D) of EU(-) = 110 pm). The tight binding of I to the EU(-) complex results from its extremely slow off rate (k(off) = 0.0008 s(-1)), which is 25,000-fold slower than substrate analogue DNA. Removal of Asp(64) and His(187), which are involved in stabilization of the cationic sugar and the anionic uracil leaving group, respectively, specifically weakens binding of I to the UDG-uracil complex by 154,000-fold, without significantly affecting substrate or product binding. These results suggest that electrostatic effects can effectively stabilize such an intermediate by at least -7 kcal/mol, without leading to anticatalytic stabilization of the substrate and products. PMID- 11859081 TI - Topography of helices 5-7 in membrane-inserted diphtheria toxin T domain: identification and insertion boundaries of two hydrophobic sequences that do not form a stable transmembrane hairpin. AB - The T domain of diphtheria toxin undergoes a low pH-induced conformational change that allows it to penetrate cell membranes. T domain hydrophobic helices 8 and 9 can adopt two conformations, one close to the membrane surface (P state) and a second in which they apparently form a transmembrane hairpin (TM state). We have now studied T domain helices 5-7, a second cluster of hydrophobic helices, using Cys-scanning mutagenesis. After fluorescently labeling a series of Cys residues, penetration into a non-polar environment, accessibility to externally added antibodies, and relative depth in the bilayer were monitored. It was found that helices 5-7 insert shallowly in the P state and deeply in the TM state. Thus, the conformational changes in helices 5-7 are both similar and somehow linked to those in helices 8 and 9. The boundaries of deeply inserting sequences were also identified. One deeply inserted segment was found to span residues 270 to 290, which overlaps helix 5, and a second spanned residues 300 to 320, which includes most of helix 6 and all of helix 7. This indicates that helices 6 and 7 form a continuous hydrophobic segment despite their separation by a Pro-containing kink. Additionally, it is found that in the TM state some residues in the hydrophilic loop between helices 5 and 6 become more highly exposed than they are in the P state. Their exposure to external solution in the TM state indicates that helices 5-7 do not form a stable transmembrane hairpin. However, helix 5 and/or helices 6 plus 7 could form transmembrane structures that are in equilibrium with non transmembrane states, or be kinetically prevented from forming a transmembrane structure. How helices 5-7 might influence the mechanism by which the T domain aids translocation of the diphtheria toxin A chain across membranes is discussed. PMID- 11859083 TI - Novel zinc-binding center and a temperature switch in the Bacillus stearothermophilus L1 lipase. AB - The bacterial thermoalkalophilic lipases optimally hydrolyze saturated fatty acids at elevated temperatures. They also have significant sequence homology with staphylococcal lipases, and both the thermoalkalophilic and staphylococcal lipases are grouped as the lipase family I.5. We report here the first crystal structure of the lipase family I.5, the structure of a thermoalkalophilic lipase from Bacillus stearothermophilus L1 (L1 lipase) determined at 2.0-A resolution. The structure is in a closed conformation, and the active site is buried under a long lid helix. Unexpectedly, the structure exhibits a zinc-binding site in an extra domain that accounts for the larger molecular size of the family I.5 enzymes in comparison to other microbial lipases. The zinc-coordinated extra domain makes tight interactions with the loop extended from the C terminus of the lid helix, suggesting that the activation of the family I.5 lipases may be regulated by the strength of the interactions. The unusually long lid helix makes strong hydrophobic interactions with its neighbors. The structural information together with previous biochemical observations indicate that the temperature mediated lid opening is triggered by the thermal dissociation of the hydrophobic interactions. PMID- 11859084 TI - A trypanosome mitochondrial RNA polymerase is required for transcription and replication. AB - Understanding mitochondrial transcription is a requisite first step toward understanding the regulation of mitochondrial gene expression in kinetoplastids. Here we report the identification and functional characterization of a mitochondrial RNA polymerase (mtRNAP) from Trypanosoma brucei, the first trans acting factor involved in kinetoplast mitochondrial transcription to be identified. Using sequences conserved among the catalytic domains of the single subunit mtRNAPs, we were able to obtain a full-length sequence for a candidate mtRNAP from T. brucei. Sequence comparison indicates that it shares homology in its catalytic domain with other single-subunit mtRNAPs, including functionally conserved residues that are identical in all single-subunit RNAPs. We used RNA interference to functionally knock out the gene product to determine whether the candidate gene represents an mtRNAP. As predicted for a mitochondrial specific RNA polymerase, reduction of the gene product resulted in a specific decrease of mitochondrial versus nuclear transcripts. Additionally, similar to the mtRNAP of other organisms, the mtRNAP characterized here is involved in replication of the mitochondrial genome. Thus, based on sequence comparison and functional studies, we have cloned an mtRNAP from trypanosomes. PMID- 11859085 TI - The effects of modifying the surface charge on the catalytic activity of a thermolysin-like protease. AB - The impact of long range electrostatic interactions on catalysis in the thermolysin-like protease from Bacillus stearothermophilus was studied by analyzing the effects of inserting or removing charges on the protein surface. Various mutations were introduced at six different positions, and double-mutant cycle analysis was used to study the extent to which mutational effects were interdependent. The effects of single point mutations on the k(cat)/K(m) were non additive, even in cases where the point mutations were located 10 A or more from the active site Zn(2+) and separated from each other by up to 25 A. This shows that catalysis is affected by large electrostatic networks that involve major parts of the enzyme. The interdependence of mutations at positions as much as 25 A apart in space also indicates that other effects, such as active site dynamics, play an important role in determining active site electrostatics. Several mutations yielded a significant increase in the activity, the most active (quadruple) mutant being almost four times as active as the wild type. In some cases the shape of the pH-activity profile was changed significantly. Remarkably, large changes in the pH-optimum were not observed. PMID- 11859086 TI - Xeroderma pigmentosum complementation group A protein (XPA) modulates RPA-DNA interactions via enhanced complex stability and inhibition of strand separation activity. AB - Replication protein A (RPA) participates in many cellular functions including DNA replication and nucleotide excision repair. A direct interaction between RPA and the xeroderma pigmentosum group A protein (XPA) facilitates the assembly of a preincision complex during the processing of DNA damage by the nucleotide excision repair pathway. We demonstrate here the formation of a ternary RPA, XPA, and duplex cisplatin-damaged DNA complex as is evident by electrophoretic supershift analysis. The RPA-XPA complex displays modest specificity for damaged versus undamaged duplex DNA, and the RPA-XPA complex displays a greater affinity for binding duplex cisplatin-damaged DNA when compared with the RPA or XPA proteins alone, consistent with previous results. Using DNA denaturation assays, we demonstrate that the role of XPA is in the stabilization of the duplex DNA structure via inhibition of the strand separation activity of RPA. Rapid kinetic analysis indicates that the bimolecular k(on) of the RPA-XPA complex is 2.5-fold faster than RPA alone for binding a duplex cisplatin-damaged DNA. The dissociation rate, k(off), of the RPA-XPA complex is slower than that of the RPA protein alone, suggesting that the XPA protein stabilizes the initial binding of RPA to duplex DNA as well as maintaining the integrity of the duplex DNA. Interestingly, XPA has no effect on the k(on) of RPA for a single-stranded 40-mer DNA. PMID- 11859087 TI - Tyrosine 537 within the Na+,K+-ATPase alpha-subunit is essential for AP-2 binding and clathrin-dependent endocytosis. AB - In renal epithelial cells endocytosis of Na(+),K(+)-ATPase molecules is initiated by phosphorylation of its alpha(1)-subunit, leading to activation of phosphoinositide 3-kinase and adaptor protein-2 (AP-2)/clathrin recruitment. The present study was performed to establish the identity of the AP-2 recognition domain(s) within the Na(+),K(+)-ATPase alpha(1)-subunit. We identified a conserved sequence (Y(537)LEL) within the alpha(1)-subunit that represents an AP 2 binding site. Binding of AP-2 to the Na(+),K(+)-ATPase alpha(1)-subunit in response to dopamine (DA) was increased in OK cells stably expressing the wild type rodent alpha-subunit (OK-WT), but not in cells expressing the Y537A mutant (OK-Y537A). DA treatment was associated with increased alpha(1)-subunit abundance in clathrin vesicles from OK-WT but not from OK-Y537A cells. In addition, this mutation also impaired the ability of DA to inhibit Na(+),K(+)-ATPase activity. Because phorbol esters increase Na(+),K(+)-ATPase activity in OK cells, and this effect was not affected by the Y537A mutation, the present results suggest that the identified motif is specifically required for DA-induced AP-2 binding and Na(+),K(+)-ATPase endocytosis. PMID- 11859088 TI - The FabR (YijC) transcription factor regulates unsaturated fatty acid biosynthesis in Escherichia coli. AB - Unsaturated fatty acid biosynthesis is a vital facet of Escherichia coli physiology and requires the expression of two genes, fabA and fabB, in the type II fatty acid synthase system. This study links the FabR (YijC) transcription factor to the regulation of unsaturated fatty acid content through the regulation of fabB gene expression. The yijC (fabR) gene was deleted by replacement with a selectable cassette, and the resulting strains (fabR::kan) possessed significantly elevated levels of unsaturated fatty acids, particularly cis vaccenate, in their membrane phospholipids. The altered fatty acid composition was observed in the fabR::kan fabF1 double mutant pinpointing fabB as the condensing enzyme responsible for the increased cis-vaccenate production. The fabR::kan strains had 4- to 8-fold higher levels of fabB and a 2- to 3-fold increase in fabA transcripts as judged by Northern blotting, Affymetrix array analysis, and real-time PCR. FabR did not regulate the enzymes of fatty acid beta oxidation. The elevated level of fabB mRNA was reflected by higher condensing enzyme activity in fabR::kan fabF1 double mutants. Thus, FabR functions as a repressor that potently controls the expression of the fabB gene, which in turn, modulates the physical properties of the membrane by altering the level of unsaturated fatty acid production. PMID- 11859089 TI - Design of a novel peptide inhibitor of HIV fusion that disrupts the internal trimeric coiled-coil of gp41. AB - The pre-hairpin intermediate of gp41 from the human immunodeficiency virus (HIV) is the target for two classes of fusion inhibitors that bind to the C-terminal region or the trimeric coiled-coil of N-terminal helices, thereby preventing formation of the fusogenic trimer of hairpins. Using rational design, two 36 residue peptides, N36(Mut(e,g)) and N36(Mut(a,d)), were derived from the parent N36 peptide comprising the N-terminal helix of the gp41 ectodomain (residues 546 581 of HIV-1 envelope), characterized by analytical ultracentrifugation and CD, and assessed for their ability to inhibit HIV fusion using a quantitative vaccinia virus-based fusion assay. N36(Mut(e,g)) contains nine amino acid substitutions designed to disrupt interactions with the C-terminal region of gp41 while preserving contacts governing the formation of the trimeric coiled-coil. N36(Mut(a,d)) contains nine substitutions designed to block formation of the trimeric coiled-coil but retains residues that interact with the C-terminal region of gp41. N36(Mut(a,d)) is monomeric, is largely random coil, does not interact with the C34 peptide derived from the C-terminal region of gp41 (residues 628-661), and does not inhibit fusion. The trimeric coiled-coil structure is therefore a prerequisite for interaction with the C-terminal region of gp41. N36(Mut(e,g)) forms a monodisperse, helical trimer in solution, does not interact with C34, and yet inhibits fusion about 50-fold more effectively than the parent N36 peptide (IC(50) approximately 308 nm versus approximately 16 microm). These results indicate that N36(Mut(e,g)) acts by disrupting the homotrimeric coiled-coil of N-terminal helices in the pre-hairpin intermediate to form heterotrimers. Thus N36(Mut(e,g)) represents a novel third class of gp41 targeted HIV fusion inhibitor. A quantitative model describing the interaction of N36(Mut(e,g)) with the pre-hairpin intermediate is presented. PMID- 11859090 TI - Effect of point mutations in the N terminus of the lentivirus lytic peptide-1 sequence of human immunodeficiency virus type 1 transmembrane protein gp41 on Env stability. AB - To understand the role of the lentivirus lytic peptide-1 region of the human immunodeficiency virus type 1 transmembrane glycoprotein (gp) 41 in viral infection, we examined the effects on virus replication of single amino acid deletions spanning this region in an infectious provirus of the HXB2 strain. Among the mutants analyzed, only the deletion of one of the two adjacent valine residues located at positions 832 and 833 (termed the Delta 833 mutant for simplicity) greatly reduced the steady-state, cell-associated levels of the Env precursor and gp120, as opposed to the wild-type virus. The altered Env phenotype resulted in severely impaired virus infectivity and gp120 incorporation into this mutant virion. Analyses of additional mutants with deletions at Ile-830, Ala-836, and Ile-840 demonstrated that the Delta 830 mutant exhibited the most significant inhibitory effect on Env steady-state expression. These results indicate that the N terminus of the lentivirus lytic peptide-1 region is critical for Env steady state expression. Among the mutant viruses encoding Env proteins in which residues Val-832 and Val-833 were individually substituted by nonconserved amino acids Ala, Ser, or Pro, which were expected to disrupt the alpha-helical structure in the increasingly severe manner of Pro > Ser > Ala, only the 833P mutant exhibited significantly reduced steady-state Env expression. Pulse labeling and pulse-chase studies demonstrated that the Delta 830, Delta 833, and 833P mutants of Env proteins degraded more rapidly in a time-dependent manner after biosynthesis than did the wild-type Env. The results indicate that residue 830 and 833 mutations are likely to induce a conformational change in Env that targets the mutant protein for cellular degradation. Our study has implications about the structural determinants located at the N terminus of the lentivirus lytic peptide-1 sequence of gp41 that affect the fate of Env in virus-infected cells. PMID- 11859091 TI - Reference: CD Antigens 2002. PMID- 11859092 TI - Cutting Edge: Transmembrane phosphoprotein Csk-binding protein/phosphoprotein associated with glycosphingolipid-enriched microdomains as a negative feedback regulator of mast cell signaling through the FcepsilonRI. AB - Tyrosine phosphorylation in the cytoplasmic domains of FcepsilonRI by the Src family kinase Lyn initiates a signaling cascade leading to mast cell activation. In this study, we show that a recently identified transmembrane protein, Csk binding protein (Cbp), also known as phospoprotein associated with glycosphingolipid-enriched microdomains (PAG), negatively regulates FcepsilonRI signaling. In rat basophilic leukemia (RBL)-2H3 cells, the levels of tyrosine phosphorylation of Cbp/PAG and its association with Csk, a negative regulator for Lyn, significantly elevate immediately after aggregation of FcepsilonRI. An overexpression of Cbp/PAG in RBL-2H3 cells inhibits FcepsilonRI-mediated cell activation. This is accompanied with decreased levels of tyrosine phosphorylation of FcepsilonRI, association of FcepsilonRI with Lyn, and FcepsilonRI-associated tyrosine kinase activity. These findings combined with the fact that Cbp/PAG, Lyn, and aggregated FcepsilonRI are localized to lipid rafts, suggest that upon FcepsilonRI aggregation Cbp/PAG down-regulates the receptor-associated Lyn activity through relocating Csk to rafts, thereby efficiently mediating feedback inhibition of FcepsilonRI signaling. PMID- 11859093 TI - Cutting Edge: Internalization of transduced E-selectin by cultured human endothelial cells: comparison of dermal microvascular and umbilical vein cells and identification of a phosphoserine-type di-leucine motif. AB - Persistent E-selectin expression on human dermal microvascular endothelial cells (HDMEC), believed to mediate skin-specific T cell homing, results from a slow rate of surface protein internalization after cytokine induction. Following transduction of unactivated HDMEC with E-selectin cDNA, the rate of internalization was largely independent of increasing levels of surface protein expression, leading to prolonged t(1/2) values of over 4 h, comparable to that observed following cytokine induction. In HUVEC, the rate of internalization increased with surface expression level, leading to an essentially constant t(1/2) of under 2 h. Thus, the internalization process rather than cytokine responsiveness or E-selectin structure underlies the difference in endothelial cell behavior. Mutational analysis of the cytoplasmic region demonstrated a role for a di-leucine-type motif involving I588 and L589 but not for a putative tyrosine-type motif. Control of E-selectin surface expression appears to be phosphoserine dependent, since alanine but not aspartic acid substitution for S581 slows E-selectin internalization. PMID- 11859094 TI - Cutting edge: Attenuated experimental autoimmune encephalomyelitis in eta 1/osteopontin-deficient mice. AB - Recent studies indicate that early T lymphocyte activation 1 (Eta-1), also known as osteopontin, is a cytokine contributing to the development of Th1 immunity. In the present report, the role of Eta-1 in experimental autoimmune encephalomyelitis (EAE), a disease associated with Th1 immunity, was examined by analysis of disease progression in Eta-1-deficient (Eta-1-/-) mice. Although incidence and onset of peptide-induced EAE were found to be similar in Eta-1-/- and Eta-1+/+ mice, Eta-1-/- mice displayed significantly lower mean maximal clinical score and faster recovery without spontaneous relapses. Accordingly, decreased inflammatory infiltration and demyelination were observed in the spinal cords of Eta-1-/- mice. Furthermore, in comparison to Eta-1+/+, Eta-1-/- CD4+ T cells had reduced expression of IFN-gamma and TNF-alpha upon ex vivo restimulation. Taken together, these results suggest that Eta-1 may sustain autoimmune responses by assisting in maintenance of Th1 immunity during EAE. PMID- 11859095 TI - Transitional B lymphocyte subsets operate as distinct checkpoints in murine splenic B cell development. AB - Signaling through the Ag receptor is required for peripheral B lymphocyte maturation and maintenance. Defects in components of the B cell receptor (BCR) signalosome result in developmental blocks at the transition from immature (heat stable Ag (HSA)(high)) to mature (HSA(low)) B cells. Recent studies have subdivided the immature, or transitional, splenic B cells into two subsets, transitional 1 (T1) and transitional 2 (T2) cells. T1 and T2 cells express distinct surface markers and are located in distinct anatomic locations. In this report, we evaluated the BCR signaling capacity of T1 and T2 B cell subsets. In response to BCR engagement, T2 cells rapidly entered cell cycle and resisted cell death. In contrast, T1 cells did not proliferate and instead died after BCR stimulation. Correlating with these results, T2 cells robustly induced expression of the cell cycle regulator cyclin D2 and the antiapoptotic factors A1/Bfl-1 and Bcl-x(L) and exhibited activation of Akt. In contrast, T1 cells failed to up regulate these markers. BCR stimulation of T2 cells also led to down-regulation of CD21 and CD24 (HSA) expression, resulting in a mature B cell phenotype. In addition, T2 cells from Bruton's tyrosine kinase-deficient Xid mice failed to generate these proliferative and survival responses, suggesting a requirement for the BCR signalosome specifically at the T2 stage. Taken together, these data clearly demonstrate that T2 immature B cells comprise a discrete developmental subset that mediates BCR-dependent proliferative, prosurvival, and differentiation signals. Their distinct BCR-dependent responses suggest unique roles for T1 vs T2 cells in peripheral B cell selection. PMID- 11859096 TI - The kinetics of in vivo priming of CD4 and CD8 T cells by dendritic/tumor fusion cells in MUC1-transgenic mice. AB - Previous work has demonstrated that dendritic/tumor fusion cells induce potent antitumor immune responses in vivo and in vitro. However, little is known about the migration and homing of fusion cells after s.c. injection or the kinetics of CD4+ and CD8+ T cell activation. In the present study, fluorescence-labeled dendritic/MUC1-positive tumor fusion cells (FC/MUC1) were injected s.c. into MUC1 transgenic mice. The FC/MUC1 migrated to draining lymph nodes and were closely associated with T cells in a pattern comparable with that of unfused dendritic cells. Immunization of MUC1-transgenic mice with FC/MUC1 resulted in proliferation of T cells and induced MUC1-specific CD8+ CTL. Moreover, CD4+ T cells activated by FC/MUC1 were multifunctional effectors that produced IL-2, IFN gamma, IL-4, and IL-10. These findings indicate that both CD4+ and CD8+ T cells can be primed in vivo by FC/MUC1 immunization. PMID- 11859097 TI - The dendritic cell-specific adhesion receptor DC-SIGN internalizes antigen for presentation to T cells. AB - Dendritic cells (DCs) capture Ags or viruses in peripheral tissue to transport them to lymphoid organs to induce cellular T cell responses. Recently, a DC specific C-type lectin was identified, DC-specific ICAM-grabbing non-integrin (DC SIGN), that functions as cell adhesion receptor mediating both DC migration and T cell activation. DC-SIGN also functions as an HIV-1R that captures HIVgp120 and facilitates DC-induced HIV transmission of T cells. Internalization motifs in the cytoplasmic tail of DC-SIGN hint to a function of DC-SIGN as endocytic receptor. In this study we demonstrate that on DCs DC-SIGN is rapidly internalized upon binding of soluble ligand. Mutating a putative internalization motif in the cytoplasmic tail reduces ligand-induced internalization. Detailed analysis using ratio fluorescence imaging and electron microscopy showed that DC-SIGN-ligand complexes are targeted to late endosomes/lysosomes. Moreover, ligands internalized by DC-SIGN are efficiently processed and presented to CD4+ T cells. The distinct pattern of expression of C-type lectins on DCs in situ and their nonoverlapping Ag recognition profile hint to selective functions of these receptors to allow a DC to recognize a wide variety of Ags and to process these to induce T cell activation. These data point to a novel function of the adhesion receptor DC-SIGN as an efficient DC-specific Ag receptor that can be used as a target to induce viral and antitumor immunity. PMID- 11859098 TI - Receptor-facilitated antigen presentation requires the recruitment of B cell linker protein to Igalpha. AB - Ags that cross-link the B cell Ag receptor are preferentially and rapidly delivered to the MHC class II-enriched compartment for processing into peptides and subsequent loading onto MHC class II. Proper sorting of Ag/receptor complexes requires the recruitment of Syk to the phosphorylated immunoreceptor tyrosine based activation motif tyrosines of the B cell Ag receptor constituent Igalpha. We postulated that the Igalpha nonimmunoreceptor tyrosine-based activation motif tyrosines, Y(176) and Y(204), contributed to receptor trafficking. Igalpha(YDeltaF(176,204))/Igbeta receptors were targeted to late endosomes, but were excluded from the vesicle lumen and could not facilitate the presentation of Ag to T cells. Subsequent analysis demonstrated that phosphorylation of Y(176)/Y(204) recruited the B cell linker protein, Vav, and Grb2. Reconstitution of Igalpha(YDeltaF(176,204))/Igbeta with the B cell linker protein rescued both receptor-facilitated Ag presentation and entry into the MHC class II-enriched compartment. Thus, aggregation accelerates receptor trafficking by recruiting two separate signaling modules required for transit through sequential checkpoints. PMID- 11859099 TI - Fucosyltransferase VII-deficient mice with defective E-, P-, and L-selectin ligands show impaired CD4+ and CD8+ T cell migration into the skin, but normal extravasation into visceral organs. AB - The first step of leukocyte extravasation, leukocyte rolling, is mediated by E-, P-, and L-selectins. Mice deficient for alpha-1,3-fucosyltransferase VII (FucTVII)(-/-) are characterized by deficiency of E-, P-, and L-selectin ligand activity. This model system was used to evaluate the role of the interactions of selectins with their ligands in T and B cell responses. In the present study, FucTVII(-/-) mice showed reduced CD4+ T cell-mediated contact hypersensitivity reactions of the ears to FITC as well as reduced CD8+ T cell-mediated delayed type hypersensitivity reactions of the footpads against lymphocytic choriomeningitis virus infection. As Langerhans cell migration to local lymph nodes as well as CD4+ and CD8+ T cell induction were found to be normal, the afferent arm of these reactions was not impaired. The reduced inflammatory reactions of the skin were due to inefficient lymphocyte extravasation into the skin. In contrast, extravasation of CD4+ and CD8+ T cells into visceral organs, such as the ovaries or the brain, was not impaired in FucTVII(-/-) mice. Elimination of vaccinia virus and of lymphocytic choriomeningitis virus from ovaries and brain, as well as elimination of tumor cells from several visceral organs was normal. Thus, interactions of selectins with their ligands are important for lymphocyte homing into the skin, but not for lymphocyte extravasation into visceral organs. PMID- 11859100 TI - Hyperproliferative response of a monoclonal memory CD8 T cell population is characterized by an increased frequency of clonogenic precursors. AB - Strong memory T cell responses result partly from the selection of Ag-specific clones during immunization. In this study, we show that a monoclonal CD8 T cell population expressing a unique TCR is heterogeneous in terms of clonogenic potential following activation under optimal conditions. More importantly, the frequency of clonogenic cells is strongly increased among Ag-experienced cells, indicating that these cells were either generated or selected during the in vivo primary response. Moreover, strong proliferative responses of primed cells result from this enhanced frequency, as proliferating naive and primed cells display the same cycling parameters, i.e., lag time and intermitotic interval. Hence, these results suggest that the clonogenic potential of individual cells is imprinted before Ag encounter and that clonogenic precursors are selected or generated following in vivo activation. PMID- 11859101 TI - Efficient delivery of T cell epitopes to APC by use of MHC class II-specific Troybodies. AB - A major objective in vaccine development is the design of reagents that give strong, specific T cell responses. We have constructed a series of rAb with specificity for MHC class II (I-E). Each has one of four different class II restricted T cell epitopes genetically introduced into the first C domain of the H chain. These four epitopes are: 91-101 lambda2(315), which is presented by I E(d); 110-120 hemagglutinin (I-E(d)); 323-339 OVA (I-A(d)); and 46-61 hen egg lysozyme (I-A(k)). We denote such APC-specific, epitope-containing Ab "Troybodies." When mixed with APC, all four class II-specific Troybodies were approximately 1,000 times more efficient at inducing specific T cell activation in vitro compared with nontargeting peptide Ab. Furthermore, they were 1,000 10,000 times more efficient than synthetic peptide or native protein. Conventional intracellular processing of the Troybodies was required to load the epitopes onto MHC class II. Different types of professional APC, such as purified B cells, dendritic cells, and macrophages, were equally efficient at processing and presenting the Troybodies. In vivo, class II-specific Troybodies were at least 100 times more efficient at targeting APC and activating TCR-transgenic T cells than were the nontargeting peptide Ab. Furthermore, they were 100-100,000 times more efficient than synthetic peptide or native protein. The study shows that class II-specific Troybodies can deliver a variety of T cell epitopes to professional APC for efficient presentation, in vitro as well as in vivo. Thus, Troybodies may be useful as tools in vaccine development. PMID- 11859102 TI - Defective Fas ligand expression and activation-induced cell death in the absence of IL-2-inducible T cell kinase. AB - The Tec family tyrosine kinase, IL-2-inducible T cell kinase (Itk), plays an important role in TCR signaling. Studies of T cells from Itk-deficient mice have demonstrated that Itk is critical for the activation of phospholipase-Cgamma1, leading to calcium mobilization in response to TCR stimulation. This biochemical defect results in reduced IL-2 production by Itk-deficient T cells. To further characterize the downstream effects of the Itk deficiency, we crossed Itk-/- mice to a TCR-transgenic line and examined T cell responses to stimulation by peptide plus APC. These studies show that Itk is required for maximal activation of early growth responses 2 and 3 and Fas ligand transcription after TCR stimulation. These transcriptional defects lead to reduced activation-induced cell death of stimulated Itk-/- T cells, both in vitro and in vivo. Together these studies define an important role for Itk in TCR signaling, leading to cytokine gene expression and activation-induced cell death. PMID- 11859103 TI - Correlation of tissue distribution, developmental phenotype, and intestinal homing receptor expression of antigen-specific B cells during the murine anti rotavirus immune response. AB - The intestinal homing receptor, alpha(4)beta(7), helps target lymphocytes to Peyer's patches (PP) and intestinal lamina propria (ILP). We have previously shown that protective immunity to rotavirus (RV), an intestinal pathogen, resides in memory B cells expressing alpha(4)beta(7). In this study, using a novel FACS assay, we have directly studied the phenotype of B cells that express surface RV specific Ig during the in vivo RV immune response. During primary infection, RV specific B cells first appear as large IgD(-)B220(low)alpha(4)beta(7)(-)and alpha(4)beta(7)(+) cells (presumptive extrafollicular, Ab-secreting B cells), and then as large and small IgD(-)B220(high)alpha(4)beta(7)(-)cells (presumptive germinal center B cells). The appearance of B cells with the phenotype of large IgD(-)B220(low)alpha(4)beta(7)(+) cells in PP and most notably in mesenteric lymph nodes coincides with the emergence of RV-specific Ab-secreting cells (ASC) in the ILP. Thus, these B lymphocytes are good candidates for the migratory population giving rise to the RV-specific ASC in the ILP. RV-specific long-term memory B cells preferentially accumulate in PP and express alpha(4)beta(7). Nine months after infection most RV-specific IgA ASC are found in PP and ILP and at lower frequency in bone marrow and spleen. This study is the first to follow changes in tissue-specific homing receptor expression during Ag-specific B cell development in response to a natural host, tissue-specific pathogen. These results show that alpha(4)beta(7) is tightly regulated during the Ag-specific B cell response to RV and is expressed concurrently with the specific migration of memory and effector B cells to intestinal tissues. PMID- 11859104 TI - Evidence for the extrathymic origin of intestinal TCRgammadelta(+) T cells in normal rats and for an impairment of this differentiation pathway in BB rats. AB - The BB rat lyp mutation, one of its diabetes susceptibility genes, is responsible for a 5-fold decrease in the number of peripheral TCRalphabeta(+) T cells. In this study we show that TCRgammadelta(+) T cells are virtually undetectable among splenic T cells and intestinal intraepithelial T lymphocytes (IEL) of BB rats, while they account for 3 and 30% of these two T cell populations, respectively, in normal animals. It has been shown that murine IEL expressing TCRgammadelta develop extrathymically. We determined whether this is the case in rats. Athymic radiation chimeras reconstituted with normal hemopoietic precursors were devoid of donor-derived TCRalphabeta(+) T cells and TCRgammadelta(+) splenocytes but contained a normal number of TCRgammadelta(+) IEL, suggesting that in unmanipulated rats some of the TCRgammadelta(+) IEL may have an extrathymic origin. This was further supported by the observation that RAG1 transcripts are present in IEL of unmanipulated animals. No T cells developed in chimeras reconstituted with BB hemopoietic precursors, demonstrating that the BB rat lyp mutation inhibits both intrathymic and extrathymic development of TCRgammadelta(+) T cells. PMID- 11859105 TI - Administration of an antigen at a high dose generates regulatory CD4+ T cells expressing CD95 ligand and secreting IL-4 in the liver. AB - Ags administered orally at a high dose are absorbed in immunogenic forms and perfuse the liver, which raises a question regarding the relevance of hepatic lymphocyte activation to the systemic hyporesponsiveness against the ingested Ag. Oral administration of 100 mg of OVA to the mice led to massive cell death of OVA specific (KJ1-26+)CD4+ T cells by Fas-Fas ligand (FasL)-mediated apoptosis in the liver, which was associated with the emergence of hepatic KJ1-26+CD4+ T cells expressing FasL. Hepatic CD4+ T cells in OVA-fed mice secreted large amounts of IL-4, IL-10, and TGF-beta(1) upon restimulation in vitro and inhibited T cell proliferation. Adoptive transfer of these hepatic CD4+ T cells to naive mice and subsequent antigenic challenge led to suppression of T cell proliferation as well as IgG Ab responses to OVA; this effect was mostly abrogated by a blocking Ab to FasL. i.p. administration of an Ag at a high dose also generated hepatic CD4+FasL+ T cells with similar cytokine profile as T cells activated by oral administration of Ags at a high dose. Finally, we did not see an increase in FasL+ cells in the hepatic CD4+Vbeta8+ T cell subset of MRL/lpr/lpr mice given staphylococcal enterotoxin B, indicating the requirement for Fas-mediated signals. These hepatic CD4+FasL+ regulatory cells may explain the tolerogenic property of the liver and play roles in systemic hyporesponsiveness induced by an Ag administered at a high dose. PMID- 11859106 TI - The nonclassical major histocompatibility complex molecule Qa-2 protects tumor cells from NK cell- and lymphokine-activated killer cell-mediated cytolysis. AB - The cytotoxic activity of NK cells is regulated by class I MHC proteins. Although much has been learned about NK recognition of class I autologous targets, the mechanisms of NK self-tolerance are poorly understood. To examine the role of a nonpolymorphic, ubiquitously expressed class Ib Ag, Q9, we expressed it on class I-deficient and NK-sensitive B78H1 melanoma. Presence of this Qa-2 family member on tumor cells partially protected targets from lysis by bulk lymphokine activated killer (LAK) cells. H-2K(b)-expressing B78H1 targets also reduced LAK cell activity, while H-2D(b) offered no protection. Importantly, blocking with F(ab')(2) specific for Q9 or removal of this GPI-attached molecule by phospholipase C cleavage restored killing to the level of vector-transfected cells. Experiments with LAK cells derived from H2(b) SCID and B6 mice established that NK1.1(+)TCR(-) NK and NK1.1(+)TCR(+) LAK cells were the prevalent cytolytic populations inhibitable by Q9. Treatment of mice with poly(I:C) also resulted in generation of Q9-regulated splenic cytotoxicity. LAK cells from different mouse strains responded to Q9, suggesting that the protective effect of this molecule is not detectably influenced by Ly49 polymorphisms or the presence/absence of Q9 in NK-harboring hosts. We propose that Q9 expressed on melanoma cells serves as a ligand for yet unidentified NK inhibitory receptor(s) expressed on NK1.1(+) NK/T cells. PMID- 11859107 TI - Positive effects of glucocorticoids on T cell function by up-regulation of IL-7 receptor alpha. AB - Despite the effects of glucocorticoids on immune function, relatively little is known about glucocorticoid-inducible genes and how their products may regulate lymphocyte function. Using DNA microarray technology to analyze gene expression in PBMC from healthy donors, we identified IL-7Ralpha as a glucocorticoid inducible gene. This observation was confirmed at the mRNA and protein levels. Conversely, TCR signaling decreased IL-7Ralpha expression, and the relative strength of signaling between these two receptors determined the final IL-7Ralpha levels. The up-regulation of IL-7Ralpha by glucocorticoids was associated with enhanced IL-7-mediated signaling and function. Moreover, IL-7-mediated inhibition of apoptosis at increasing concentrations of glucocorticoids is consistent with enhanced cell sensitivity to IL-7 following glucocorticoid exposure. These observations provide a mechanism by which glucocorticoids may have a positive influence on T cell survival and function. PMID- 11859108 TI - Efficient in vivo priming of specific cytotoxic T cell responses by neonatal dendritic cells. AB - In early life, a high susceptibility to infectious diseases as well as a poor capacity to respond to vaccines are generally observed as compared with observations in adults. The mechanisms underlying immune immaturity have not been fully elucidated and could be due to the immaturity of the T/B cell responses and/or to a defect in the nature and quality of Ag presentation by the APC. This prompted us to phenotypically and functionally characterize early life murine dendritic cells (DC) purified from spleens of 7-day-old mice. We showed that neonatal CD11c(+) DC express levels of costimulatory molecules and MHC molecules similar to those of adult DC and are able to fully maturate after LPS activation. Furthermore, we demonstrated that neonatal DC can efficiently take up, process, and present Ag to T cells in vitro and induce specific CTL responses in vivo. Although a reduced number of these cells was observed in the spleen of neonatal mice as compared with adults, this study clearly shows that neonatal DC have full functional capacity and may well prime Ag-specific naive T cells in vivo. PMID- 11859109 TI - Functional expression of the chemokine receptor CCR5 on virus epitope-specific memory and effector CD8+ T cells. AB - Because the chemokine receptor CCR5 is expressed on Th1 CD4(+) cells, it is important to investigate the expression and function of this receptor on other T cells involved in Th1 immune responses, such as Ag-specific CD8(+) T cells, which to date have been only partially characterized. Therefore, we analyzed the expression and function of CCR5 on virus-specific CD8+ T cells identified by HLA class I tetramers. Multicolor flow cytometry analysis demonstrated that CCR5 is expressed on memory (CD28+CD45RA-) and effector (CD28-CD45RA- and CD28-CD45RA+) CD8+ T cells but not on naive (CD28+CD45RA+) CD8+ T cells. CCR5 expression was much lower on two effector CD8+ T cells than on memory CD8+ T cells. Analysis of CCR7 and CCR5 expression on the different types of CD8+ T cells showed that memory CD8+ T cells have three phenotypic subsets, CCR5+CCR7-, CCR5+CCR7+, and CCR5-CCR7+, while naive and effector CD8+ T cells have CCR5-CCR7+ and CCR5+CCR7- phenotypes, respectively. These results suggest the following sequence for differentiation of memory CD8+ T cells: CCR5-CCR7+-->CCR5+CCR7+-->CCR5+CCR7-. CCR5+CD8+ T cells effectively migrated in response to RANTES, suggesting that CCR5 plays a critical role in the migration of Ag-specific effector and differentiated memory CD8+ T cells to inflammatory tissues and secondary lymphoid tissues. This is in contrast to CCR7, which functions as a homing receptor in migration of naive and memory CD8+ T cells to secondary lymphoid tissues. PMID- 11859110 TI - Lipid raft heterogeneity in human peripheral blood T lymphoblasts: a mechanism for regulating the initiation of TCR signal transduction. AB - Lateral mobility and spatial organization of proteins within the plasma membrane are likely to mediate the initial events coordinating T cell activation. Lipid rafts, distinct cholesterol/sphingolipid-rich membrane microdomains, provide a mechanism for this regulation by concentrating or excluding signaling proteins. We demonstrate in peripheral blood T cell lymphoblasts that immediate early phosphotyrosine signal transduction through the TCR complex is functionally dependent on a distinct population of lipid rafts. Specifically, cholesterol extraction destabilizes the membrane microdomains containing Lck, while the rafts containing the adapter protein linker for activation of T cells remain intact. Heterogeneity in the partitioning of these proteins in resting cells was confirmed by immunoelectron microscopy. After T cell activation, both Lck and the linker for activation of T cells colocalize to 50-100 nm microdomains in the plasma membrane, indicating that sequestration of these proteins into distinct lipid rafts may function to regulate the initiation of T cell signal transduction. PMID- 11859111 TI - Antigen-antibody immune complexes empower dendritic cells to efficiently prime specific CD8+ CTL responses in vivo. AB - Dendritic cells (DCs) require a maturation signal to acquire efficient CTL priming capacity. In vitro FcgammaR-mediated internalization of Ag-Ab immune complexes (ICs) can induce maturation of DCs. In this study, we show that IC induced DC maturation in vitro enables DCs to prime peptide-specific CD8+ CTLs in vivo, independently of CD4+ Th cells. Importantly, OVA/anti-OVA IC-treated DCs not only primed CD8+ CTLs to an exogenously loaded peptide nonrelated to OVA, but also efficiently primed CTLs against the dominant CTL epitope derived from the OVA Ag present in the ICs. Our studies show that ICs fulfill a dual role in priming of CD8+ CTL responses to exogenous Ags: enhancement of Ag uptake by DCs and activation of DCs, resulting in "license to kill." These findings indicate that the presence of specific Abs can crucially affect the induction of cytotoxic cellular responses. PMID- 11859112 TI - HIV antigens can induce TGF-beta(1)-producing immunoregulatory CD8+ T cells. AB - HIV-infected individuals may progressively lose both HIV-specific and unrelated CTL responses despite the high number of circulating CD8+ T cells. In this study, we report that approximately 25% of HIV+ donors produced TGF-beta(1) in response to stimulation with HIV proteins or peptides. The production of TGF-beta(1) was sufficient to significantly reduce the IFN-gamma response of CD8+ cells to both HIV and vaccinia virus proteins. Ab to TGF-beta reversed the suppression. We found the source of the TGF-beta(1) to be predominantly CD8+ cells. Different peptide pools stimulated TGF-beta(1) and IFN-gamma in the same individual. The TGF-beta(1) secreting cells have distinct peptide specificity from the IFN-gamma producing cells. This represents an important mechanism by which an HIV-specific response can nonspecifically suppress both HIV-specific and unrelated immune responses. PMID- 11859113 TI - Cyclooxygenase-2-issued prostaglandin e(2) enhances the production of endogenous IL-10, which down-regulates dendritic cell functions. AB - PGE(2) is a well-known immunomodulator produced in the immune response by APCs, such as dendritic cells (DCs), the most potent APC of the immune system. We investigated the PGE(2) biosynthetic capacity of bone marrow-derived DC (BM-DC) and the effects of PG on the APC. We observed that BM-DC produce PGE(2) and other proinflammatory mediators, such as leukotriene B(4) and NO, after LPS exposure. Constitutively present in BM-DC, cyclooxygenase (COX)-1 did not contribute significantly to the total pool of PGE(2) compared with the LPS-induced COX-2 produced PGE(2). Treatment of BM-DC with exogenous PGE(2) induced the production of large amounts of IL-10 and less IL-12p70. In addition, selective inhibition of COX-2, but not COX-1, was followed by significant decrements in PGE(2) and IL-10, a concomitant restoration of IL-12 production, and an enhancement of DC stimulatory potential. In contrast, we found no demonstrable role for leukotriene B(4) or NO. In view of the potential of PGE(2) to stimulate IL-10, we examined the possibility that the suppressive effect of PGE(2) is mediated via IL-10. We found that exogenous IL-10 inhibits IL-12p70 production in the presence of NS 398, a COX-2 selective inhibitor, while the inhibitory effects of PGE(2) were totally reversed by anti-IL-10. We conclude that COX-2-mediated PGE(2) up regulates IL-10, which down-regulates IL-12 production and the APC function of BM DC. PMID- 11859114 TI - Expression of thrombospondin in TGFbeta-treated APCs and its relevance to their immune deviation-promoting properties. AB - APCs deployed within iris/ciliary body are responsible for promoting anterior chamber-associated immune deviation following injection of Ag into the eye. TGFbeta-2, a constituent of the ocular microenvironment, converts conventional APCs that are pulsed with Ag into cells that induce immune deviation when injected into naive mice. TGFbeta-2-treated APCs under-express IL-12 and CD40, and over-express active TGFbeta. We have examined transcriptional changes within macrophage hybridoma no. 59, which promotes Th1 cell differentiation, and TGFbeta 2-treated no. 59 as well as macrophage hybridoma no. 63, both of which induce immune deviation similar to anterior chamber-associated immune deviation. Immune deviation-inducing hybridomas up-regulated expression of thrombospondin, TGFbeta, IFN-alpha and beta, murine macrophage elastase, and macrophage-inflammatory protein-2 genes, while down-regulating expression of the genes for NF-kappaB and CD40. Based on the known properties of these gene products, a model is proposed in which these gene products, alone and through interacting signaling pathways, confer upon conventional APCs the capacity to create and surround themselves with an immunomodulatory microenvironment. The model proposes that the pleiotropic effects of thrombospondin are primarily responsible for creating this microenvironment that is stabile, rich in active TGFbeta and IFN-alpha and beta, deficient in IL-12, and chemoattractant via macrophage-inflammatory protein-2 for NK T cells. It is further proposed that presentation of Ag to T cells in this microenvironment leads to their differentiation into regulatory cells that suppress Th1 cell-dependent immunogenic inflammation. PMID- 11859115 TI - Tracking the immunoregulatory mechanisms active during allograft tolerance. AB - Immunoregulatory mechanisms dependent on regulatory CD4+ T cells are believed to be critical in the maintenance of peripheral tolerance to allografts. However, a detailed characterization of the effects of these regulatory T cells has been hampered by the absence of a simple means to track and study them. In this work we provide evidence that in a murine model of islet transplantation the interactions between alloaggressive and regulatory T cells can be studied in vitro and in vivo at the single-cell level. The observations made in both an in vitro coculture system and an in vivo CFSE-based adoptive transfer model indicate that lymphocytes from tolerant allograft recipients 1) proliferate weakly to donor strain allogeneic cells but vigorously to third-party strain cells; and 2) suppress the proliferation of naive syngeneic CD4+ and CD8+ T cells to donor tissue in a cell dose- and Ag-specific manner. These effects depend on the presence of CD4+CD25+ T cells and are neutralized by anti-CTLA4 mAb or rIL-2. The principal effect of anti-CTLA4 is directed against the naive, not regulatory, T cell population. These results can be replicated in vivo by transferring lymphocyte populations into transplant recipients, proving that the graft protecting actions of regulatory T cells are blunted by a rise in the number of allodestructive T cells (pool size model) and depend on the presence of CD4+CD25+ T cells and the integrity of the CTLA4/B7 pathway. PMID- 11859116 TI - IL-12 induces monocyte IL-18 binding protein expression via IFN-gamma. AB - IL-18 is a Th1 cytokine that synergizes with IL-12 and IL-2 in the stimulation of lymphocyte IFN-gamma production. IL-18 binding protein (IL-18BP) is a recently discovered inhibitor of IL-18 that is distinct from the IL-1 and IL-18 receptor families. In this report we show that IL-18BPa, the IL-18BP isoform with the highest affinity for IL-18, was strongly induced by IL-12 in human PBMC. Other Th1 cytokines, including IFN-gamma, IL-2, IL-15, and IL-18, were also capable of augmenting IL-18BPa expression. In contrast, IL-1alpha, IL-1beta, TNF-alpha, IFN gamma-inducible protein-10, and Th2 cytokines such as IL-4 and IL-10 did not induce IL-18BPa. Although monocytes were found to be the primary source of IL 18BPa, the induction of IL-18BPa by IL-12 was mediated through IFN-gamma derived predominantly from NK cells. IL-18BPa production was observed in cancer patients receiving recombinant human IL-12 and correlated with the magnitude of IFN-gamma production. The IFN-gamma/IL-18BPa negative feedback loop identified in this study may be capable of broadly controlling immune activation by cytokines that synergize with IL-18 to induce IFN-gamma and probably plays a key role in the modulation of both innate and adaptive immunity. PMID- 11859117 TI - The apoptotic protease-activating factor 1-mediated pathway of apoptosis is dispensable for negative selection of thymocytes. AB - Negative selection is a process to delete potentially autoreactive clones in developing thymocytes. Programmed cell death or apoptosis is thought to play an important role in this selection process. In this study, we investigated the role of apoptotic protease-activating factor 1 (Apaf1), a mammalian homologue of CED 4, in programmed cell death during the negative selection in thymus. There was no developmental abnormality in thymocytes from newborn Apaf1(-/-) mice in terms of CD4 and CD8 expression pattern and thymocyte number. Clonal deletion by endogenous male H-Y Ag of Apaf1-deficient thymocytes with transgenic expression of H-Y Ag-specific TCRs (H-Y Tg/Apaf1(-/-) thymocytes) was normally observed in lethally irradiated wild-type mice reconstituted with fetal liver-derived hemopoietic stem cells. Clonal deletion induced in vitro by a bacterial superantigen was also normal in fetal thymic organ culture. Thus, Apaf1-mediated pathway of apoptosis is dispensable for the negative selection of thymocytes. However, H-Y Tg/Apaf1(-/-) thymocytes showed partial resistance to H-Y peptide induced deletion in vitro as compared with H-Y Tg/Apaf1(+/-) thymocytes, implicating the Apaf1-mediated apoptotic pathway in the negative selection in a certain situation. In addition, the peptide-induced deletion was still observed in H-Y Tg/Apaf1(-/-) thymocytes in the presence of a broad spectrum caspase inhibitor, z-VAD-fmk, suggesting the presence of caspase-independent cell death pathway playing roles during the negative selection. We assume that mechanisms for the negative selection are composed of several cell death pathways to avoid failure of elimination of autoreactive clones. PMID- 11859118 TI - The role of the CPNKEKEC sequence in the beta(2) subunit I domain in regulation of integrin alpha(L)beta(2) (LFA-1). AB - The alpha(L) I (inserted or interactive) domain of integrin alpha(L)beta(2) undergoes conformational changes upon activation. Recent studies show that the isolated, activated alpha(L) I domain is sufficient for strong ligand binding, suggesting the beta(2) subunit to be only indirectly involved. It has been unclear whether the activity of the alpha(L) I domain is regulated by the beta(2) subunit. In this study, we demonstrate that swapping the disulfide-linked CPNKEKEC sequence (residues 169-176) in the beta(2) I domain with a corresponding beta(3) sequence, or mutating Lys(174) to Thr, constitutively activates alpha(L)beta(2) binding to ICAM-1. These mutants do not require Mn(2+) for ICAM-1 binding and are insensitive to the inhibitory effect of Ca(2+). We have also localized a component of the mAb 24 epitope (a reporter of beta(2) integrin activation) in the CPNKEKEC sequence. Glu(173) and Glu(175) of the beta(2) I domain are identified as critical for mAb 24 binding. Because the epitope is highly expressed upon beta(2) integrin activation, it is likely that the CPNKEKEC sequence is exposed or undergoes conformational changes upon activation. Deletion of the alpha(L) I domain did not eliminate the mAb 24 epitope. This confirms that the alpha(L) I domain is not critical for mAb 24 binding, and indicates that mAb 24 detects a change expressed in part in the beta(2) subunit I domain. These results suggest that the CPNKEKEC sequence of the beta(2) I domain is involved in regulating the alpha(L) I domain. PMID- 11859119 TI - Evolution of Ig DNA sequence to target specific base positions within codons for somatic hypermutation. AB - Ig variable (V) region genes are subjected to a somatic hypermutation process as B lymphocytes participate in immune reactions to protein Ags. Although little is known regarding the mechanism of mutagenesis, a consistent hierarchy of trinucleotide target preferences is evident. Analysis of trinucleotide regional distributions predicted and we now empirically confirm the surprising finding that the framework 2 region of kappa V region genes is highly mutable despite its importance to the structural integrity and function of the Ab molecule. Interestingly, much of this mutability appears to be focused on the third codon position where synonymous substitutions are most likely to occur. We also observed a trend for high predicted mutability for codon positions 1 and 2 in complementarity-determining regions. Consequently, amino acid replacements should occur at a higher rate in complementarity-determining regions than in framework regions due to the distribution and subsequent targeting of microsequences by the mutation mechanism. Our results reveal a subtle tier of V region gene evolution in which DNA sequence has been molded to direct mutations to specific base positions within codons in a manner that minimizes damage and maximizes the benefits of the somatic hypermutation process. PMID- 11859120 TI - Allelic polymorphism synergizes with variable gene content to individualize human KIR genotype. AB - Killer Ig-like receptor (KIR) genes are a multigene family on human chromosome 19. KIR genes occur in various combinations on different haplotypes. Additionally, KIR genes are polymorphic. To examine how allelic polymorphism diversifies KIR haplotypes with similar or identical combinations of KIR genes, we devised methods for discriminating alleles of KIR2DL1, -2DL3, -3DL1, and 3DL2. These methods were applied to 143 individuals from 34 families to define 98 independent KIR haplotypes at the allele level. Three novel 3DL2 alleles and a chimeric 3DL1/3DL2 sequence were also identified. Among the A group haplotypes were 22 different combinations of 2DL1, 2DL3, 3DL1, and 3DL2 alleles. Among the B group haplotypes that were unambiguously determined were 15 distinct haplotypes involving 9 different combinations of KIR genes. A and B haplotypes both exhibit strong linkage disequilibrium (LD) between 2DL1 and 2DL3 alleles, and between 3DL1 and 3DL2 alleles. In contrast, there was little LD between the 2DL1/2DL3 and 3DL1/3DL2 pairs that define the two halves of the KIR gene complex. The synergistic combination of allelic polymorphism and variable gene content individualize KIR genotype to an extent where unrelated individuals almost always have different KIR types. This level of diversity likely reflects strong pressure from pathogens on the human NK cell response. PMID- 11859121 TI - A change in the structure of Vbeta chromatin associated with TCR beta allelic exclusion. AB - To investigate chromatin control of TCR beta rearrangement and allelic exclusion, we analyzed TCR beta chromatin structure in double negative (DN) thymocytes, which are permissive for TCR beta recombination, and in double positive (DP) thymocytes, which are postallelic exclusion and nonpermissive for Vbeta to DbetaJbeta recombination. Histone acetylation mapping and DNase I sensitivity studies indicate Vbeta and DbetaJbeta segments to be hyperacetylated and accessible in DN thymocytes. However, they are separated from each other by hypoacetylated and inaccessible trypsinogen chromatin. The transition from DN to DP is accompanied by selective down-regulation of Vbeta acetylation and accessibility. The level of DP acetylation and accessibility is minimal for five of six Vbeta segments studied but remains substantial for one. Hence, the observed changes in Vbeta chromatin structure appear sufficient to account for allelic exclusion of many Vbeta segments. They may contribute to, but not by themselves fully account for, allelic exclusion of others. PMID- 11859122 TI - Transgenic expression of the p16(INK4a) cyclin-dependent kinase inhibitor leads to enhanced apoptosis and differentiation arrest of CD4-CD8- immature thymocytes. AB - In the thymus, T cell development proceeds by successive steps of differentiation, expansion, and selection. Control of thymocyte proliferation is critical to insure the full function of the immune system and to prevent T cells from transformation. Deletion of the cell cycle inhibitor p16(INK4a) is frequently observed in human T cell neoplasias and, in mice, gene targeted inactivation of the Ink4a locus enhances thymocyte expansion and predisposes mutant animal to tumorigenesis. Here, we investigate the mechanism by which p16(Ink4a) controls thymocyte development by analyzing transgenic mice expressing the human p16(INK4a) into the T cell lineage. We show that forced expression of p16(INK4a) in thymocytes blocked T cell differentiation at the early CD4-CD8-CD3 CD25+ stage without significantly affecting the development of gammadelta T cells. Pre-TCR function was mimicked by the induction of CD3 signaling in thymocytes of recombinase activating gene (RAG)-2-deficient mice (RAG-2(-/-)). Upon anti-CD3epsilon treatment in vivo, p16(INK4a)-expressing RAG-2(-/-) thymocytes were not rescued from apoptosis, nor could they differentiate. Our data demonstrate that expression of p16(INK4a) prevents the pre-TCR-mediated expansion and/or survival of differentiating thymocytes. PMID- 11859123 TI - Evidence for the murine IgH mu locus acting as a hot spot for intrachromosomal homologous recombination. AB - Homologous recombination accomplishes the exchange of genetic information between two similar or identical DNA duplexes. It can occur either by gene conversion, a process of unidirectional genetic exchange, or by reciprocal crossing over. Homologous recombination is well known for its role in generating genetic diversity in meiosis and, in mitosis, as a DNA repair mechanism. In the immune system, the evidence suggests a role for homologous recombination in Ig gene evolution and in the diversification of Ab function. Previously, we reported the occurrence of homologous recombination between repeated, donor and recipient alleles of the Ig H chain mu gene C (Cmu) region residing at the Ig mu locus in mouse hybridoma cells. In this study, we constructed mouse hybridoma cell lines bearing Cmu region heteroalleles to learn more about the intrachromosomal homologous recombination process. A high frequency of homologous recombination (gene conversion) was observed for markers spanning the entire recipient Cmu region, suggesting that recombination might initiate at random sites within the Cmu region. The Cmu region heteroalleles were equally proficient as either conversion donors or recipients. Remarkably, when the same Cmu heteroalleles were tested for recombination in ectopic genomic positions, the mean frequency of gene conversion was reduced by at least 65-fold. These results are consistent with the murine IgH mu locus behaving as a hot spot for intrachromosomal homologous recombination. PMID- 11859124 TI - Xenopus laevis Stromal cell-derived factor 1: conservation of structure and function during vertebrate development. AB - Transmembrane signaling of the CXC chemokine stromal cell-derived factor-1 (SDF 1) is mediated by CXCR4, a G protein-coupled receptor initially identified in leukocytes and shown to serve as a coreceptor for the entry of HIV into lymphocytes. Characterization of SDF-1- and CXCR4-deficient mice has revealed that SDF-1 and CXCR4 are of vital developmental importance. To study the role of the SDF-1/CXCR4-chemokine/receptor system as a regulator of vertebrate development, we isolated and characterized a cDNA encoding SDF-1 of the lower vertebrate Xenopus laevis (xSDF-1). Recombinant xSDF-1 was produced in insect cells, purified, and functionally characterized. Although xSDF-1 is only 64-66% identical with its mammalian counterparts, it is indistinguishable from human (h)SDF-1alpha in terms of activating both X. laevis CXCR4 and hCXCR4. Thus, both xSDF-1 and hSDF-1alpha promoted CXCR4-mediated activation of heterotrimeric G(i2) in a cell-free system and induced release of intracellular calcium ions in and chemotaxis of intact lymphoblastic cells. Analysis of the time course of xSDF-1 mRNA expression during Xenopus embryogenesis revealed a tightly coordinated regulation of xSDF-1 and X. laevis CXCR4. xSDF-1 mRNA was specifically detected in the developing CNS, incipient sensory organs, and the embryonic heart. In Xenopus, CXCR4 mRNA appears to be absent from the heart anlage, but present in neural crest cells. This observation suggests that xSDF-1 expressed in the heart anlage may attract cardiac neural crest cells expressing CXCR4 to migrate to the primordial heart to regulate both septation of the cardiac outflow tract and differentiation of the myocardium during early heart development. PMID- 11859126 TI - Endotoxin-neutralizing antimicrobial proteins of the human placenta. AB - Microbial colonization and infection of placental tissues often lead to adverse pregnancy outcomes such as preterm birth, a leading cause of neonatal morbidity and mortality. The fetal membranes of the placenta, a physical and active barrier to microbial invasion, encapsulate the fetus and secure its intrauterine environment. To examine the innate defense system of the human placenta, antimicrobial peptides were isolated from the fetal membranes of human placenta and characterized biochemically. Two salt-resistant antimicrobial host proteins were purified to homogeneity using heparin-affinity and reversed-phase HPLC. Characterization of these proteins revealed that they are identical to histones H2A and H2B. Histones H2A and H2B showed dose-dependent inhibition of the endotoxin activity of LPS and inhibited this activity by binding to and therefore blocking both the core and lipid A moieties of LPS. Consistent with a role for histones in the establishment of placental innate defense, histones H2A and H2B were highly expressed in the cytoplasm of syncytiotrophoblasts and amnion cells, where the histone proteins were localized mainly to the epithelial surface. Furthermore, culturing of amnion-derived WISH cells led to the constitutive release of histone H2B, and histones H2A and H2B contribute to bactericidal activity of amniotic fluid. Our studies suggest that histones H2A and H2B may endow the epithelium of the placenta with an antimicrobial and endotoxin neutralizing barrier against microorganisms that invade this immune-privileged site. PMID- 11859125 TI - Transient association of Ku with nuclear substrates characterized using fluorescence photobleaching. AB - The autoantigen Ku, composed of subunits Ku70 and Ku86, is necessary for repair of DNA double-strand breaks by nonhomologous end joining. Similarly, Ku participates in repair of DNA double-strand breaks that occur during V(D)J recombination, and it is therefore required for the development of B and T lymphocytes. Although previous studies have identified the DNA-binding activities of Ku, little is known concerning its dynamics, such as the mobility of Ku in the nucleus and its rate of association with substrates. To address this question, fluorescence photobleaching experiments were performed using HeLa cells and B cells expressing a green fluorescent protein (GFP) fusion construct of either Ku70 or Ku86. The results show that Ku moves rapidly throughout the nucleus even following irradiation of the cells. However, the rate of diffusion of Ku was approximately 100-fold slower than that predicted from its size. Association of Ku-GFP with a filamentous nuclear structure was also evident, and nuclear extraction experiments suggest that this represents nuclear matrix. A central domain of Ku70 containing its DNA-binding and heterodimerization regions and its nuclear localization signal shows that this alone is sufficient for the observed mobility of Ku70-GFP and its association with nuclear matrix. These data suggest the mobility of Ku is characterized by a transient, high flux association with nuclear substrates that includes both DNA and the nuclear matrix and may represent a mechanism for repair of double-strand breaks using the nuclear matrix as a scaffold. PMID- 11859127 TI - Txk, a member of nonreceptor tyrosine kinase of Tec family, acts as a Th1 cell specific transcription factor and regulates IFN-gamma gene transcription. AB - Precise mechanisms responsible for Th1 cell activation and differentiation are not fully elucidated. We have recently reported that Txk, a member of Tec family nonreceptor tyrosine kinase, is expressed on Th1/Th0 cells, and Txk regulates specifically IFN-gamma gene expression. In this study, we found that Txk bound to IFN-gamma promoter region. Txk transfection increased transcriptional activity of IFN-gamma promoter plus luciferase constructs severalfold, including IFN-gamma promoter -538, -208, and -53. IFN-gamma promoter -39 was refractory to the Txk transfection. The actual site to which Txk bound was the element consisting of 53 and -39 bp from the transcription start site of human IFN-gamma gene, a site distinct from several previously characterized binding sites. We found that the entire -53/-39 region was necessary for the binding to and function of Txk, because mutant promoter oligoDNA that contained contiguous five base substitutions dispersed throughout the -53/-39 inhibited the binding, and the mutant promoters did not respond to the Txk transfection. Similar sequences of this element are found within the 5' flanking regions of several Th1 cell associated protein genes. Thus, Txk is expressed on Th1/Th0 cells with the IFN gamma production and acts as a Th1 cell-specific transcription factor. PMID- 11859128 TI - Epitope recognition by diverse antibodies suggests conformational convergence in an antibody response. AB - Crystal structures of distinct mAbs that recognize a common epitope of a peptide Ag have been determined and analyzed in the unbound and bound forms. These Abs display dissimilar binding site structures in the absence of the Ag. The dissimilarity is primarily expressed in the conformations of complementarity determining region H3, which is responsible for defining the epitope specificity. Interestingly, however, the three Abs exhibit similar complementarity-determining region conformations in the Ag binding site while recognizing the common epitope, indicating that different pathways of binding are used for Ag recognition. The epitope also exhibits conformational similarity when bound to each of these Abs, although the peptide Ag was otherwise flexible. The observed conformational convergence in the epitope and the Ag binding site was facilitated by the plasticity in the nature of interactions. PMID- 11859129 TI - IL-10 is critical for host resistance and survival during gastrointestinal helminth infection. AB - Resistance to many intestinal nematodes is dependent on the induction of polarized type 2 cytokine responses, whereas type 1 responses can exacerbate these infections. The contributions of IL-4 and IL-13 to the development of resistance have been well described for a variety of intestinal parasites; however, the role of IL-10 has not been previously investigated. In this study we infected IL-10-, IL-10/IL-4-, IL-10/IL-12-, IL-4-, and IL-12-deficient mice with Trichuris muris to determine whether IL-10 contributes to the development of immunity. Interestingly, T. muris-infected IL-10-, IL-4-, and IL-10/IL-4 deficient mice failed to expel the parasite, and animals deficient in IL-10 displayed marked morbidity and mortality. In contrast, double IL-10/IL-12 deficient mice were completely resistant and mounted a highly polarized type 2 cytokine response, demonstrating that the increased susceptibility of IL-10 deficient mice was dependent on IL-12. Further study suggested that the susceptibility of IL-10- and IL-10/IL-4-deficient mice was probably attributable to a marked increase in type 1 cytokine production in those animals. The mortality observed in T. muris-infected IL-10- and IL-10/IL-4-deficient mice correlated with increased inflammation, loss of Paneth cells, and absence of mucus in the cecum. Interestingly, survival was enhanced in T. muris-infected IL 10/IL-4-deficient mice if a broad spectrum antibiotic was administered, suggesting that an outgrowth of opportunistic bacteria was contributing to the high degree of morbidity and mortality. Overall, these studies reveal a critical role for IL-10 in the polarization of Th2 responses, development of resistance during T. muris infection, and maintenance of barrier function in the colon. PMID- 11859130 TI - Dendritic cells transduced with protein antigens induce cytotoxic lymphocytes and elicit antitumor immunity. AB - Dendritic cell (DC)-based vaccines are being developed for treatment of patients with cancer, in part because DC are potent inducers of CD8(+) CTL. DC MHC class I:antigenic peptide complexes that are required for CTL elicitation are most often generated by incubating DC with peptides or by transfecting (or transducing) DC with cDNAs (or viral vectors) that encode protein Ags. The former approach is feasible when MHC class I Ags and relevant peptides are known. The latter approach may be hampered by inefficient DC transfection (transduction) and/or difficulties associated with preparation and use of viral vectors. Herein we demonstrate that a bacterial recombinant model tumor-associated Ag (OVA) that contains the HIV TAT protein transduction domain (PTD) was readily engineered and purified, efficiently transduced murine lymphocytes and DC, and was processed by proteasomes for MHC class I-restricted presentation to CTL. In addition, PTD containing rOVA was processed and presented by DC to CD4 T cells as efficiently as native OVA or rOVA lacking the PTD. PTD-OVA-transduced DC induced CTL in vivo in a Th cell-independent fashion and vaccinated against OVA-expressing tumors. In contrast, rOVA lacking the PTD did not enter the DC MHC class I presentation pathway and DC treated with this protein did not prime OVA-specific CTL in vivo. Treatment of mice harboring clinically apparent OVA-expressing tumors with PTD OVA-transduced DC resulted in tumor regression in some animals. This straightforward vaccination strategy may translate into DC-based treatments for patients with cancer and other serious diseases. PMID- 11859131 TI - Identification of BING-4 cancer antigen translated from an alternative open reading frame of a gene in the extended MHC class II region using lymphocytes from a patient with a durable complete regression following immunotherapy. AB - Multiple human cancer Ags have been identified, although little is known concerning which would be most effectively used in cancer immunotherapy. To gain insight into the selection of appropriate Ags, the immunologic reactivity of a patient who had a durable complete regression of melanoma metastases was measured. PBMCs were directly cloned using the monoclonal anti-CD3 Ab OKT3 and IL 2 without any bias introduced by previous culture. A lymphocyte clone recognized a previously unknown shared melanoma Ag that was identified as the BING-4 protein encoded in a gene-rich region of the extended class II MHC. The HLA-A2-restricted BING-4 immunodominant peptide was translated from a 10-aa-long alternative open reading frame. In vitro sensitization against this peptide generated lymphocytes reactive against HLA-A2(+) melanomas. Real-time semiquantitative RT-PCR analysis revealed that 8 of 15 melanoma cell lines overexpressed BING-4, and this correlated with recognition by lymphocytes. Overexpression was not found in normal tissues or other tumor types. Thus, BING-4 represents another candidate Ag for possible use in the immunotherapy of patients with melanoma. PMID- 11859132 TI - Follicular dendritic cells and the persistence of HIV infectivity: the role of antibodies and Fcgamma receptors. AB - Large quantities of HIV are found trapped on the surface of follicular dendritic cells (FDCs), and virus persists on these cells until they ultimately die. We recently found that FDCs maintain HIV infectivity for long periods in vivo and in vitro. Because FDCs trap Ags (and virus) in the form of immune complexes and are rich in FcgammaRs, we reasoned that Ab and FcgammaRs may be required for FDC mediated maintenance of HIV infectivity. To investigate this hypothesis, HIV immune complexes were formed in vitro and incubated for increasing times with or without FDCs, after which the remaining infectious virus was determined by HIV p24 production in rescue cultures. FDCs maintained HIV infectivity in vitro in a dose-dependent manner but required the presence of specific Ab for this activity regardless of whether laboratory-adapted or primary X4 and R5 isolates were tested. In addition, Abs against either virally or host-encoded proteins on the virion permitted FDC-mediated maintenance of HIV infectivity. We found that the addition of FDCs to HIV immune complexes at the onset of culture gave optimal maintenance of infectivity. Moreover, blocking FDC-FcgammaRs or killing the FDCs dramatically reduced their ability to preserve virus infectivity. Finally, FDCs appeared to decrease the spontaneous release of HIV-1 gp120, suggesting that FDC virus interactions stabilize the virus particle, thus contributing to the maintenance of infectivity. Therefore, optimal maintenance of HIV infectivity requires both Ab against particle-associated determinants and FDC-FcgammaRs. PMID- 11859133 TI - Serpin 2a is induced in activated macrophages and conjugates to a ubiquitin homolog. AB - After i.p. infection of mice with the intracellular bacterium Mycobacterium bovis bacillus Calmette-Guerin, macrophages recovered from the peritoneal cavity display classical signs of immune activation. We have identified a member of the serine protease inhibitor (serpin) family which is highly induced in macrophages during bacillus Calmette-Guerin infection. Serpin 2a (spi2a) expression is also induced in macrophages in vivo during infection with Salmonella typhimurium and Listeria monocytogenes, and in vitro by a variety of bacteria and bacterial products. The cytokine IFN-gamma also induces spi2a expression in macrophages, and this induction is synergistic with bacterial products. We also demonstrate here that a ubiquitin homolog, IFN-stimulated gene of 15-kDa (ISG15), is strongly induced during in vitro and in vivo activation of macrophages and that it conjugates to spi2a in activated macrophages. The ISG15-spi2a conjugates were identified by tandem mass spectrometry and contained spi2a conjugated to either one or two molecules of ISG15. Whereas spi2a was induced by either bacterial products or IFN-gamma, ISG15 was induced only by bacterial products. Although many protein targets have been described for ubiquitin conjugation, spi2a is the first ISG15-modified protein to be reported. Macrophage activation is accompanied by the activation of a variety of proteases. It is of interest that a member of the serine protease inhibitor family is concomitantly induced and modified by a ubiquitin-like protein. PMID- 11859134 TI - Response to Neisseria gonorrhoeae by cervicovaginal epithelial cells occurs in the absence of toll-like receptor 4-mediated signaling. AB - Toll-like receptors (TLRs) have recently been identified as fundamental components of the innate immune response to bacterial pathogens. We investigated the role of TLR signaling in immune defense of the mucosal epithelial cells of the lower female genital tract. This site provides first line defense against microbial pathogens while remaining tolerant to a complex biosystem of resident microbiota. Epithelial cells derived from normal human vagina, ectocervix, and endocervix expressed mRNA for TLR1, -2, -3, -5, and -6. However, they failed to express TLR4 as well as MD2, two essential components of the receptor complex for LPS in phagocytes and endothelial cells. Consistent with this, endocervical epithelial cells were unresponsive to protein-free preparations of lipooligosaccharide from Neisseria gonorrhoeae and LPS from Escherichia coli. However, they were capable of responding to whole Gram-negative bacteria and bacterial lysates, as demonstrated by NF-kappaB activation and proinflammatory cytokine production. The presence of soluble CD14, a high-affinity receptor for LPS and other bacterial ligands, enhanced the sensitivity of genital tract epithelial cells to both low and high concentrations of bacteria, suggesting that soluble CD14 can act as a coreceptor for non-TLR4 ligands. These data demonstrate that the response to N. gonorrhoeae and other Gram-negative bacteria at the mucosal surface of the female genital tract occurs in the absence of endotoxin recognition and TLR4-mediated signaling. PMID- 11859135 TI - Susceptibility of IFN regulatory factor-1 and IFN consensus sequence binding protein-deficient mice to brucellosis. AB - IFN-gamma is a key cytokine controlling Brucella infection, and the diverse functions of this cytokine are mediated by IFN regulatory factors (IRFs) such as IRF-1, IRF-2, and IFN consensus sequence binding protein (ICSBP). However, the roles of these three IRFs in Brucella infection have not been investigated. The infection of each IRF-deficient mouse strain provides an opportunity to determine not only the significance of each IRF molecule but also the crucial immune components necessary for host defense during in vivo infection, because respective IRF-deficient mouse strains contain unique immunodeficient phenotypes. Brucella abortus S2308-infected IRF-1-/- mice were dead within 2 wk postinfection, while IRF-2-/- mice contained less splenic Brucella CFU than wild type mice at the early stage of infection. Infected ICSBP-/- mice maintained a plateau of splenic Brucella CFU throughout the infection. Additional infection of IL-12p40-, NO synthase 2-, and gp91(phox)-deficient mice indicates that these immune components are crucial for Brucella immunity and may contribute to the susceptibility of IRF-1-/- and ICSBP-/- mice. Immunologic and histopathological analyses of infected IRF-1-/- mice indicate that the absence of IL-12p40 induction and serious hepatic damage are involved in the death of IRF-1-/- mice. These results indicate that 1) IRF-1 and ICSBP are essential transcriptional factors for IFN-gamma-mediated protection against Brucella; 2) IL-12, reactive nitrogen intermediates, and reactive oxygen intermediates are crucial immune components against Brucella, and their absence may contribute to the susceptibility of IRF-1-/- and ICSBP-/- mice; and 3) hepatic damage caused by Brucella virulence contributes to the death of IRF-1-/- mice. PMID- 11859136 TI - Cooperation of C1q receptors and integrins in C1q-mediated endothelial cell adhesion and spreading. AB - The interaction of C1q with endothelial cells elicits a multiplicity of biologic responses. Although these responses are presumed to be mediated by the interaction of C1q with endothelial cell surface proteins, the identity of the participants is not known. In this study we examined the roles of two C1q binding proteins, cC1q-R/calreticulin and gC1q-R/p33, in C1q-mediated adhesion and spreading of human dermal microvascular endothelial cells (HDMVEC). When HDMVEC were cultured in microtiter plate wells coated with concentrations of C1q ranging from 0 to 50 microg/ml, a specific and dose-dependent adhesion and spreading was observed. The extent of adhesion and spreading was similar to the adhesion seen on collagen-coated wells. Spreading (68 +/- 12%) and to a moderate extent adhesion (47 +/- 9%) were inhibited by anti-gC1q-R mAb 60.11. Similar effects were noted with polyclonal anti-cC1q-R but not with control nonimmune IgG. The two Abs had a slight additive effect (75 +/- 13% inhibition) when mixed together in the proportion of 100 microg/ml anti-gC1q-R and 30 microg/ml anti-cC1q-R. More importantly, a 100% inhibition of spreading, but not adhesion, to C1q-coated wells was observed when HDMVEC were cultured in the presence of 30 microM of the peptide GRRGDSP but not GRRGESP. Furthermore, while anti-beta(1) integrin Ab blocked both adhesion and spreading, anti-alpha(5) integrin blocked only spreading and not adhesion. Ag capture ELISA of endothelial cell membrane proteins using polyclonal anti-gC1q-R showed the presence of not only beta(1) and alpha(5) integrins but also CD44. Taken together these results suggest that endothelial cell adhesion and spreading require the cooperation of both C1qRs and beta(1) integrins and possibly other membrane-spanning molecules. PMID- 11859137 TI - Activation of macrophage promatrix metalloproteinase-9 by lipopolysaccharide associated proteinases. AB - LPS induces an up-regulation of promatrix metalloproteinase-9 (proMMP9) gene expression in cells of the monocyte/macrophage lineage. We demonstrate here that LPS preparations are also able to activate proMMP9 made by human macrophages or THP-1 cells via LPS-associated proteinases, which cleave the N-terminal propeptide at a site or sites close to the one cleaved upon activation with organomercurial compounds. LPS-associated proteinases are serine proteinases that are able to cleave denatured collagens (gelatin) and the mammalian serine proteinase inhibitor, alpha(1)-proteinase inhibitor, thereby pushing the balance of extracellular matrix turnover even further toward degradation. A low molecular mass, low affinity inhibitor of MMP9, possibly derived from the propeptide, is generated during proMMP9 activation. However, inhibition of the LPS-associated proteinases had no effect on proMMP9 synthesis, indicating that their proteolytic activity was not required for signaling the up-regulation of the proMMP9 gene. PMID- 11859139 TI - IL-5 promotes eosinophil trafficking to the esophagus. AB - Eosinophil infiltration into the esophagus occurs in a wide range of diseases; however, the underlying pathophysiological mechanisms involved are largely unknown. We now report that the Th2 cytokine, IL-5, is necessary and sufficient for the induction of eosinophil trafficking to the esophagus. We show that transgenic mice overexpressing IL-5 under the control of a T cell (CD2) or a small intestinal enterocyte (fatty acid-binding protein) promoter have markedly increased eosinophil numbers in the esophagus. For example, esophageal eosinophil levels are 1.9 +/- 0.9 and 121 +/- 14 eosinophils/mm(2) in wild-type and CD2-IL-5 transgenic mice, respectively. Consistent with this effect being mediated by a systemic mechanism, pharmacological administration of IL-5 via a miniosmotic pump in the peritoneal cavity resulted in blood and esophageal eosinophilia. To examine the role of IL-5 in oral Ag-induced esophageal eosinophilia, eosinophilic esophagitis was induced by allergen exposure in IL-5-deficient and wild-type mice. Importantly, IL-5-deficient mice were resistant to eosinophilic esophagitis. Finally, we examined the role of eotaxin when IL-5 was overproduced in vivo. Esophageal eosinophil levels in CD2-IL-5-transgenic mice were found to decrease 15-fold in the absence of the eotaxin gene; however, esophageal eosinophil numbers in eotaxin-deficient IL-5-transgenic mice still remained higher than wild-type mice. In conclusion, these studies demonstrate a central role for IL-5 in inducing eosinophil trafficking to the esophagus. PMID- 11859138 TI - IL-4 pretreatment selectively enhances cytokine and chemokine production in lipopolysaccharide-stimulated mouse peritoneal macrophages. AB - Although well recognized for its anti-inflammatory effect on gene expression in stimulated monocytes and macrophages, IL-4 is a pleiotropic cytokine that has also been shown to enhance TNF-alpha and IL-12 production in response to stimulation with LPS. In the present study we expand these prior studies in three areas. First, the potentiating effect of IL-4 pretreatment is both stimulus and gene selective. Pretreatment of mouse macrophages with IL-4 for a minimum of 6 h produces a 2- to 4-fold enhancement of LPS-induced expression of several cytokines and chemokines, including TNF-alpha, IL-1alpha, macrophage-inflammatory protein-2, and KC, but inhibits the production of IL-12p40. In addition, the production of TNF-alpha by macrophages stimulated with IFN-gamma and IL-2 is inhibited by IL-4 pretreatment, while responses to both LPS and dsRNA are enhanced. Second, the ability of IL-4 to potentiate LPS-stimulated cytokine production appears to require new IL-4-stimulated gene expression, because it is time dependent, requires the activation of STAT6, and is blocked by the reversible protein synthesis inhibitor cycloheximide during the IL-4 pretreatment period. Finally, IL-4-mediated potentiation of TNF-alpha production involves specific enhancement of mRNA translation. Although TNF-alpha protein is increased in IL-4-pretreated cells, the level of mRNA remains unchanged. Furthermore, LPS stimulated TNF-alpha mRNA is selectively enriched in actively translating large polyribosomes in IL-4-pretreated cells compared with cells stimulated with LPS alone. PMID- 11859140 TI - A unique combination of inflammatory cytokines enhances apoptosis of thyroid follicular cells and transforms nondestructive to destructive thyroiditis in experimental autoimmune thyroiditis. AB - Treatment of cultured primary human thyroid cells with IFN-gamma and TNF-alpha uniquely allows the induction of Fas-mediated apoptosis. To investigate the role of this cytokine combination in vivo, CBA/J mice were immunized with thyroglobulin and then injected with IFN-gamma and TNF-alpha. Compared with control animals, mice treated with IFN-gamma and TNF-alpha showed significantly sustained lymphocytic infiltration in the thyroid, which was associated with the destruction of portions of the follicular architecture at wk 6 after initial immunization. Furthermore, the number of apoptotic thyroid follicular cells was increased only in the thyroids from mice treated with the IFN-gamma and TNF alpha. We also analyzed the function of the Fas pathway in vivo in cytokine treated mice by using an agonist anti-Fas Ab injected directly into the thyroid. Minimal apoptosis of thyroid epithelial cells was observed unless the mice were pretreated with IFN-gamma and TNF-alpha. These data demonstrate that this unique combination of inflammatory cytokines facilitates the apoptotic destruction of thyroid follicular cells in experimental autoimmune thyroiditis, in a manner similar to what is observed in Hashimoto's thyroiditis in humans. PMID- 11859141 TI - Nonhematopoietic expression of Janus kinase 3 is required for efficient recruitment of Th2 lymphocytes and eosinophils in OVA-induced airway inflammation. AB - Tyrosine kinases of the Janus kinase (Jak) family transduce signals from the type I and type II cytokine receptors. Jak3 is unique in this family because its expression must be induced and is predominantly limited to cells of the lymphoid and myeloid lineages. Deficient expression of Jak3 interferes with normal development and function of T, B, and NK cells. Using irradiated Jak3-deficient (Jak3-/-) mice reconstituted with normal bone marrow (Jak3-/-chimeric mice), we have investigated possible actions of Jak3 outside of the hematopoietic system. We show that efficient recruitment of inflammatory cells to the airways of OVA sensitized mice challenged with aerosolized OVA requires the expression of Jak3 in radioresistant nonhematopoietic cells. Failure to develop eosinophil predominant airway inflammation in Jak3-/- chimeric mice is not due to failure of T cell sensitization, because Jak3-/- chimeric mice showed delayed-type hypersensitivity responses indistinguishable from wild-type chimeric mice. Jak3-/ chimeric mice, however, express less endothelial-associated VCAM-1 after airway Ag challenge. Given the key role of VCAM-1 in recruitment of Th2 cells and eosinophils, our data suggest that Jak3 in airway-associated endothelial cells is required for the expression of eosinophilic airway inflammation. This requirement for nonhematopoietic expression of Jak3 represents the first demonstration of a physiological function of Jak3 outside of the lymphoid lineages. PMID- 11859142 TI - Inflammatory mediators in uveitis: differential induction of cytokines and chemokines in Th1- versus Th2-mediated ocular inflammation. AB - Ocular inflammation leads to vision loss through the destruction and scarring of delicate tissues along the visual axis. To identify inflammatory mediators involved in this process, we used real time RT-PCR to quantify the expression of mRNA transcripts of 34 cytokines, 26 chemokines, and 14 chemokine receptors at certain time points during T cell-mediated ocular inflammation. We induced disease by adoptive transfer of Ag-specific Th1 or Th2 cells into recipients expressing the target Ag in their eyes. We also compared the mediator expression patterns seen in adoptive transfer-induced inflammation with that seen in mouse eyes developing experimental autoimmune uveoretinitis. In addition, we used laser capture microdissection to examine chemokine mRNA production by both retinal pigment epithelium cells and infiltrating leukocytes in inflamed eyes. Major findings included the following: 1) Three patterns of expression of the inflammation-related molecules were seen in recipients of adoptively transferred Th cells: preferential expression in Th1 recipients, or in Th2 recipients, or similar expression in both recipient groups. 2) In experimental autoimmune uveoretinitis, the inflammatory mediator expression pattern largely paralleled that seen in Th1-induced disease. 3) Both retinal pigment epithelium and infiltrating leukocytes expressed chemokine transcripts in distinct, but overlapping patterns in inflamed eyes. 4) Interestingly, transcripts of multiple cytokines, chemokines, and chemokine receptors were constitutively expressed in high levels in mouse eyes. Seven of these molecules have not been previously associated with the eye. These data underscore the multiplicity of mediators that participate in the pathogenesis of eye inflammation and point to upstream cytokines as potential therapeutic targets. PMID- 11859143 TI - Depletion of dendritic cells, but not macrophages, in patients with sepsis. AB - Dendritic cells (DCs) are a group of APCs that have an extraordinary capacity to interact with T and B cells and modulate their responses to invading pathogens. Although a number of defects in the immune system have been identified in sepsis, few studies have examined the effect of sepsis on DCs, which is the purpose of this study. In addition, this study investigated the effect of sepsis on macrophages, which are reported to undergo apoptosis, and MHC II expression, which has been noted to be decreased in sepsis. Spleens from 26 septic patients and 20 trauma patients were evaluated by immunohistochemical staining. Although sepsis did not decrease the number of macrophages, sepsis did cause a dramatic reduction in the percentage area of spleen occupied by FDCs, i.e., 2.9 +/- 0.4 vs 0.7 +/- 0.2% in trauma and septic patients, respectively. The number of MHC II expressing cells, including interdigitating DCs, was decreased in septic, compared with trauma, patients. However, sepsis did not appear to induce a loss of MHC II expression in those B cells, macrophages, or DCs that were still present. The dramatic loss of DCs in sepsis may significantly impair B and T cell function and contribute to the immune suppression that is a hallmark of the disorder. PMID- 11859144 TI - The role of IL-12 in the induction of intravenous tolerance in experimental autoimmune encephalomyelitis. AB - Intravenous administration of autoantigen is an effective method to induce Ag specific tolerance against experimental autoimmune encephalomyelitis (EAE). IL-12 is a potent Th1 stimulator and an essential cytokine in the induction of EAE. The role of IL-12 in the induction of i.v. tolerance is not clear. In this study, we induced tolerance by i.v. administering myelin basic protein (MBP) peptide Ac1-11 (MBP1-11) in EAE. We observed significant suppression of IL-12 production by the lymph node cells of MBP1-11-injected mice. To see whether the low level of IL-12 is the cause or effect of tolerance, we administered IL-12 to the EAE mice at the time of i.v. MBP1-11 injection. Exogenous IL-12 abrogated the suppression of clinical and pathological EAE by i.v. tolerance. IL-12 blocked the suppressive effect of i.v. tolerance on the proliferative response to MBP1-11 and MBP1-11 induced production of IL-12 and IFN-gamma. Furthermore, IL-12 completely blocked the i.v. tolerance-induced type 1 T regulatory cell response. These data suggest that i.v. administration of autoantigen results in the suppression of endogenous IL-12 and the consequent switching of the immune response from an immunogenic to a tolerogenic form. PMID- 11859145 TI - Peroxisome proliferator-activated receptor-gamma agonist 15-deoxy-Delta(12,14) prostaglandin J(2) ameliorates experimental autoimmune encephalomyelitis. AB - Peroxisome proliferator-activated receptors (PPAR) are members of a nuclear hormone receptor superfamily that includes receptors for steroids, retinoids, and thyroid hormone, all of which are known to affect the immune response. Previous studies dealing with PPAR-gamma expression in the immune system have been limited. Recently, PPAR-gamma was identified in monocyte/macrophage cells. In this study we examined the role of PPAR-gamma in experimental autoimmune encephalomyelitis (EAE), an animal model for the human disease multiple sclerosis. The hypothesis we are testing is whether PPAR-gamma plays an important role in EAE pathogenesis and whether PPAR-gamma ligands can inhibit the clinical expression of EAE. Initial studies have shown that the presence of the PPAR-gamma ligand 15-deoxy-Delta(12,14)-PGJ(2) (15d-PGJ2) inhibits the proliferation of Ag specific T cells from the spleen of myelin basic protein Ac(1-11) TCR-transgenic mice. 15d-PGJ2 suppressed IFN-gamma, IL-10, and IL-4 production by both Con A- and myelin basic protein Ac(1-11) peptide-stimulated lymphocytes as determined by ELISA and ELISPOT assay. Culture of encephalitogenic T cells with 15d-PGJ2 in the presence of Ag reduced the ability of these cells to adoptively transfer EAE. Examination of the target organ, the CNS, during the course of EAE revealed expression of PPAR-gamma in the spinal cord inflammatory infiltrate. Administration of 15d-PGJ2 before and at the onset of clinical signs of EAE significantly reduced the severity of disease. These results suggest that PPAR gamma ligands may be a novel therapeutic agent for diseases such as multiple sclerosis. PMID- 11859146 TI - Airway hyperresponsiveness, late allergic response, and eosinophilia are reversed with mycobacterial antigens in ovalbumin-presensitized mice. AB - Pretreatment with mycobacterial Ags has been shown to be effective in preventing allergic airway inflammation from occurring in a mouse model. Because most asthmatics are treated after the development of asthma, it is crucial to determine whether mycobacterial Ags can reverse established allergic airway inflammation in the presensitized state. Our hypothesis, based upon our previous findings, is that mycobacteria treatment in presensitized mice will suppress the allergic airway inflammation with associated clinical correlates of established asthma, with the noted exception of factors associated with the early allergic response (EAR). BALB/c mice sensitized and challenged with OVA were evaluated for pulmonary functions during both the EAR and late allergic response, and airway hyperresponsiveness to methacholine. Following this, sensitized mice were randomized and treated with placebo or a single dose (1 x 10(5) CFUs) of bacillus Calmette-Guerin (BCG) or Mycobacterium vaccae via nasal or peritoneal injection. One week later, the mice were rechallenged with OVA and methacholine, followed by bronchoalveolar lavage (BAL) and tissue collection. Mice treated with intranasal BCG were most significantly protected from the late allergic response (p < 0.02), airway hypersensitivity (p < 0.001) and hyperreactivity (p < 0.05) to methacholine, BAL (p < 0.05) and peribronchial (p < 0.01) eosinophilia, and BAL fluid IL-5 levels (p < 0.01) as compared with vehicle-treated, sensitized controls. Intranasal M. vaccae treatment was less effective, suppressing airway hypersensitivity (p < 0.01) and BAL eosinophilia (p < 0.05). No changes were observed in the EAR, BAL fluid IL-4 levels, or serum total and Ag-specific IgE. These data suggest that mycobacterial Ags (BCG>>M. vaccae) are effective in attenuating allergic airway inflammation and associated changes in pulmonary functions in an allergen-presensitized state. PMID- 11859147 TI - An anti-idiotype vaccine elicits a specific response to N-glycolyl sialic acid residues of glycoconjugates in melanoma patients. AB - We generated the 1E10 gamma-type anti-idiotype mAb (Ab2) specific to an Ab1 mAb able to react specifically with N-glycolyl-containing gangliosides and with Ags expressed on human melanoma and breast carcinoma cells. This Ab2 mAb induced an Ab response in animal models sharing immunochemically defined idiotopes with the Ab1. The treatment of tumor-bearing mice with 1E10 mAb induced a strong antitumor activity. A clinical trial was conducted in 20 patients with advanced malignant melanoma. Patients were treated with six intradermal injections of aluminum hydroxide-precipitated 1E10 anti-Id mAb given at 2-wk intervals. Sixteen of the 17 patients who received at least four doses of the anti-Id vaccine develop Ab3 Abs capable of inhibiting Ab2 binding to Ab1 (Ab3Id+). In contrast to the incapacity of 1E10 mAb to generate Ab3 Abs with the same antigenic specificity as the Ab1 mAb in mice, a very specific and strong Ab3 response against N-glycolyl containing gangliosides was induced in 16 patients (Ab3Ag+). No evidence of serious or unexpected adverse effects has been observed in this clinical trial. 1E10 anti-Id vaccine was safe, well tolerated, and immunologically effective, with most patients being able to generate a specific immune response against 1E10 and Neu-glycolyl-GM(3) ganglioside. PMID- 11859148 TI - Naturally processed chromatin peptides reveal a major autoepitope that primes pathogenic T and B cells of lupus. AB - Major autoepitopes for pathogenic Th cells of lupus were previously found in core histones of nucleosomes by testing overlapping synthetic peptides. To detect other dominant epitopes, we eluted peptides from MHC class II molecules of a murine lupus APC line that was fed with crude chromatin. The eluted peptides were purified by reverse-phase HPLC and tested for their ability to stimulate autoimmune Th clones, and then analyzed by mass spectrometry. Amino acid sequences of stimulatory fractions revealed three new autoepitopes. Two of the epitopes were homologous to brain transcription factor BRN-3, whereas the third sequence was homologous to histone H1'(22-42). H1'(22-42) stimulated autoimmune Th cells to augment the production of pathogenic antinuclear Abs, and was much more potent than other nucleosomal epitopes in accelerating glomerulonephritis in lupus-prone (SWR x NZB)F(1) (SNF(1)) mice. Remarkably, a marked expansion of Th1 cells recognizing the H1'(22-42) epitope occurred spontaneously in SNF(1) mice very early in life. A significant proportion of H1'(22-42)-specific T cell clones cross-reacted with one or more core histone epitopes, but not with epitopes in other lupus autoantigens. The H1'(22-42) epitope was also recognized by autoimmune B cells, and with the onset of lupus nephritis, serum autoantibodies to the H1'(22-42) epitope become increasingly cross-reactive with nuclear autoantigens. Convergence of T and B cell epitopes in H1'(22-42) and its ability to elicit a cross-reactive response make it a highly dominant epitope that could be targeted for therapy and for tracking autoimmune T and B cells. PMID- 11859150 TI - Adhesion-mediated intracellular redistribution of c-Fas-associated death domain like IL-1-converting enzyme-like inhibitory protein-long confers resistance to CD95-induced apoptosis in hematopoietic cancer cell lines. AB - Evasion of immune surveillance is a key step in malignant progression. Interactions between transformed hematopoietic cells and their environment may initiate events that confer resistance to apoptosis and facilitate immune evasion. In this report, we demonstrate that beta(1) integrin-mediated adhesion to fibronectin inhibits CD95-induced caspase-8 activation and apoptosis in hematologic tumor cell lines. This adhesion-dependent inhibition of CD95-mediated apoptosis correlated with enhanced c-Fas-associated death domain-like IL-1 converting enzyme-like inhibitory protein-long (c-FLIP(L)) cytosolic solubility compared with nonadhered cells. Cytosolic c-FLIP(L) protein preferentially associated with cytosolic Fas-associated death domain protein (FADD) and localized to the death-inducing signal complex after CD95 ligation in adherent cells. The incorporation of c-FLIP(L) in the death-inducing signal complex prevented procaspase-8 processing and activation of the effector phase of apoptosis. Adhesion to fibronectin increased c-FLIP(L) cytosolic solubility and availability for FADD binding by redistributing c-FLIP(L) from a preexisting membrane-associated fraction. Increased cytosolic availability of c-FLIP(L) for FADD binding was not related to increased levels of RNA or protein synthesis. These data show that adhesion of anchorage-independent cells to fibronectin provides a novel mechanism of resistance to CD95-mediated programmed cell death by regulating the cellular localization and availability of c-FLIP(L). PMID- 11859149 TI - C1q deficiency and autoimmunity: the effects of genetic background on disease expression. AB - Gene-targeted C1q-deficient mice have been shown to develop a syndrome reminiscent of human systemic lupus erythematosus with antinuclear Abs and proliferative glomerulonephritis. Initial phenotypic analysis conducted in (129 x C57BL/6) hybrid mice showed that background genes were a significant factor for the full expression of the autoimmune disease. To assess the contribution of background genes in the expression of the autoimmune phenotype, the disrupted C1qa gene was backcrossed for seven generations onto C57BL/6 and MRL/Mp(+/+) strains. These were intercrossed with C57BL/6.lpr/lpr and MRL/Mp-lpr/lpr strains to generate C1q-deficient substrains. In C1q-deficient C57BL/6 mice, no evidence of an autoimmune phenotype was found, and C1q deficiency in both the C57BL/6.lpr/lpr and MRL/Mp-lpr/lpr strains did not modify the autoimmune phenotype observed in wild-type controls. However, in C1q-deficient MRL/Mp(+/+) animals an acceleration of both the onset and the severity of antinuclear Abs and glomerulonephritis was seen. Disease was particularly pronounced in females, which developed severe crescentic glomerulonephritis accompanied by heavy proteinuria. In addition, the C1q-deficient MRL/Mp(+/+) mice had an impairment in the phagocytic clearance of apoptotic cells in vivo. These data demonstrate that the expression of autoimmunity in C1q-deficient mice is strongly influenced by other background genes. The work also highlights the potential value of the C1q deficient MRL/Mp(+/+) strain as a tool with which to dissect further the underlying mechanisms of the autoimmune syndrome associated with C1q deficiency. PMID- 11859151 TI - A single-chain class II MHC-IgG3 fusion protein inhibits autoimmune arthritis by induction of antigen-specific hyporesponsiveness. AB - T cells play a central role in many autoimmune diseases. A method to specifically target the function of autoreactive T cell clones would avoid the global immunosuppression associated with current therapies. To develop a molecule capable of inhibiting autoreactive T cell responses in vivo, single-chain peptide I-A-IgG3 fusion proteins were constructed and expressed in both mammalian and insect cells. The fusion proteins were designed with an IgG3 Fc moiety to make them divalent, allowing TCR cross-linking, while lacking FcR binding and costimulation. The fusion proteins stimulated T cell hybridomas in vitro in a peptide-specific, MHC-restricted manner but failed to do so in soluble form. In vivo administration of an I-A(q) fusion protein, containing an immunodominant collagen II peptide, significantly delayed the onset and reduced the severity of collagen-induced arthritis in DBA/1 mice by induction of Ag-specific hyporesponsiveness. Such fusion proteins may be useful to study novel therapeutic approaches for T cell-mediated autoimmune diseases. PMID- 11859153 TI - Detecting energy emissions from a rotating black hole. AB - The rotational energy of a black hole surrounded by a torus is released through several channels. We have determined that a minor fraction of the energy is released in baryon-poor outflows from a differentially rotating open magnetic flux tube, and a major fraction of about eta/2 is released in gravitational radiation by the torus with angular velocity eta similar 0.2 to 0.5 relative to that of the black hole. We associate the energy emitted in baryon-poor outflows with gamma-ray bursts. The remaining fraction is released in torus winds, thermal emissions, and (conceivably) megaelectron-volt neutrino emissions. The emitted gravitational radiation can be detected by gravitational wave experiments and provides a method for identifying Kerr black holes in the Universe. PMID- 11859152 TI - Thiol antioxidants inhibit the adjuvant effects of aerosolized diesel exhaust particles in a murine model for ovalbumin sensitization. AB - Although several epidemiological studies indicate a correlation between exposure to ambient particulate matter and adverse health effects in humans, there is still a fundamental lack of understanding of the mechanisms involved. We set out to test the hypothesis that reactive oxygen species are involved in the adjuvant effects of diesel exhaust particles (DEP) in a murine OVA sensitization model. First, we tested six different antioxidants, N-acetylcysteine (NAC), bucillamine (BUC), silibinin, luteolin, trolox (vitamin E), and ascorbic acid, for their ability to interfere in DEP-mediated oxidative stress in vitro. Of the six agents tested, only the thiol antioxidants, BUC and NAC, were effective at preventing a decrease in intracellular reduced glutathione:glutathione disulfide ratios, protecting cells from protein and lipid oxidation, and preventing heme oxygenase 1 expression. Therefore, we selected the thiol antioxidants for testing in the murine OVA inhalation sensitization model. Our data demonstrate that NAC and BUC effectively inhibited the adjuvant effects of DEP in the induction of OVA specific IgE and IgG1 production. Furthermore, NAC and BUC prevented the generation of lipid peroxidation and protein oxidation in the lungs of OVA- plus DEP-exposed animals. These findings indicate that NAC and BUC are capable of preventing the adjuvant effects of inhaled DEP and suggest that oxidative stress is a key mechanistic component in the adjuvant effect of DEP. Antioxidant treatment strategies may therefore serve to alleviate allergic inflammation and may provide a rational basis for treating the contribution of particulate matter to asthmatic disease. PMID- 11859154 TI - Structural insights into group II intron catalysis and branch-site selection. AB - Group II self-splicing introns catalyze autoexcision from precursor RNA transcripts by a mechanism strikingly similar to that of the spliceosome, an RNA protein assembly responsible for splicing together the protein-coding parts of most eukaryotic pre-mRNAs. Splicing in both cases initiates via nucleophilic attack at the 5' splice site by the 2' OH of a conserved intron adenosine residue, creating a branched (lariat) intermediate. Here, we describe the crystal structure at 3.0 A resolution of a 70-nucleotide RNA containing the catalytically essential domains 5 and 6 of the yeast ai5gamma group II self-splicing intron, revealing an unexpected two-nucleotide bulged structure around the branch-point adenosine in domain 6. PMID- 11859155 TI - Structure of HP1 chromodomain bound to a lysine 9-methylated histone H3 tail. AB - The chromodomain of the HP1 family of proteins recognizes histone tails with specifically methylated lysines. Here, we present structural, energetic, and mutational analyses of the complex between the Drosophila HP1 chromodomain and the histone H3 tail with a methyllysine at residue 9, a modification associated with epigenetic silencing. The histone tail inserts as a beta strand, completing the beta-sandwich architecture of the chromodomain. The methylammonium group is caged by three aromatic side chains, whereas adjacent residues form discerning contacts with one face of the chromodomain. Comparison of dimethyl- and trimethyllysine-containing complexes suggests a role for cation-pi and van der Waals interactions, with trimethylation slightly improving the binding affinity. PMID- 11859156 TI - The problematic rise of Archean oxygen. PMID- 11859157 TI - Public health preparedness. PMID- 11859158 TI - Scientific community. Universities review policies for onsite classified research. PMID- 11859159 TI - Climate change. More science and a carrot, not a stick. PMID- 11859160 TI - Animal welfare. Senate says no to new rodent rules. PMID- 11859161 TI - AIDS research. Longtime rivalry ends in collaboration. PMID- 11859162 TI - Microbial genomics. TIGR begins assault on the anthrax genome. PMID- 11859163 TI - Cloning. Carbon-copy clone is the real thing. PMID- 11859164 TI - Cancer research. Obstacle for promising cancer therapy. PMID- 11859165 TI - Microbiology. Weight of the world on microbes' shoulders. PMID- 11859166 TI - Mass extinctions. No 'darkness at noon' to do in the dinosaurs? PMID- 11859167 TI - Analytical chemistry. New test could speed bioweapon detection. PMID- 11859168 TI - Astronomy. Glimpsing the post-Hubble universe. PMID- 11859170 TI - Astronomy. A hot new camera--and a chilly revival. PMID- 11859169 TI - Astronomy. NGST aims for big advance by going back in time. PMID- 11859172 TI - Astronomy. The tumultuous teens of Supernova 1987A. PMID- 11859171 TI - Neuroscience. How neurons know that it's c-c-c-c-cold outside. PMID- 11859174 TI - Seismology. Working up to the next big one. PMID- 11859173 TI - Ecology. Disciplines team up to take the pulse of Tampa Bay. PMID- 11859175 TI - American Association for the Advancement of Science annual meeting. Human gene count on the rise. PMID- 11859176 TI - American Association for the Advancement of Science annual meeting. A whale of a chain reaction. PMID- 11859177 TI - Retrospective. Scientist, mentor, colleague, and friend--Eleanore Butz (1909 2002). PMID- 11859178 TI - Public health. Is safe mail worth the price? PMID- 11859180 TI - Paleoclimate. Cycles, cycles everywhere. PMID- 11859179 TI - Neurobiology. TRK makes the retrograde. PMID- 11859181 TI - Ecology. Sizing up the shape of life. PMID- 11859182 TI - Cosmology. Tales of singularities. PMID- 11859183 TI - Immunology. The immunological synapse--a multitasking system. PMID- 11859184 TI - Enzymology. A moving story. PMID- 11859185 TI - Plants compared to animals: the broadest comparative study of development. AB - If the last common ancestor of plants and animals was unicellular, comparison of the developmental mechanisms of plants and animals would show that development was independently invented in each lineage. And if this is the case, comparison of plant and animal developmental processes would give us a truly comparative study of development, which comparisons merely among animals, or merely among plants, do not-because in each of these lineages, the fundamental mechanisms are similar by descent. Evidence from studies of developmental mechanisms in both kingdoms, and data from genome-sequencing projects, indicate that development evolved independently in the lineages leading to plants and to animals. PMID- 11859186 TI - Extracellular DNA required for bacterial biofilm formation. PMID- 11859187 TI - A rational chemical synthesis of C60. AB - Isolable quantities of C60, the smallest stable fullerene, have been synthesized in 12 steps from commercially available starting materials by rational chemical methods. A molecular polycyclic aromatic precursor bearing chlorine substituents at key positions forms C60 when subjected to flash vacuum pyrolysis at 1100 degrees C. No other fullerenes are formed as by-products. The methods we have developed for the target-specific synthesis of fullerenes, applied here to a synthesis of C60, should make possible the directed laboratory preparation of other fullerenes as well, including those not accessible by graphite vaporization. PMID- 11859188 TI - Array-based electrical detection of DNA with nanoparticle probes. AB - A DNA array detection method is reported in which the binding of oligonucleotides functionalized with gold nanoparticles leads to conductivity changes associated with target-probe binding events. The binding events localize gold nanoparticles in an electrode gap; silver deposition facilitated by these nanoparticles bridges the gap and leads to readily measurable conductivity changes. An unusual salt concentration-dependent hybridization behavior associated with these nanoparticle probes was exploited to achieve selectivity without a thermal-stringency wash. Using this method, we have detected target DNA at concentrations as low as 500 femtomolar with a point mutation selectivity factor of approximately 100,000:1. PMID- 11859189 TI - Efficient near-infrared polymer nanocrystal light-emitting diodes. AB - Conjugated polymers and indium arsenide-based nanocrystals were used to create near-infrared plastic light-emitting diodes. Emission was tunable from 1 to 1.3 micrometers--a range that effectively covers the short-wavelength telecommunications band--by means of the quantum confinement effects in the nanocrystals. The external efficiency value (photons out divided by electrons in) is approximately 0.5% (that is, >1% internal) and is mainly limited by device architecture. The near-infrared emission did not overlap the charge-induced absorption bands of the polymer. PMID- 11859190 TI - Otolith delta 18O record of mid-Holocene sea surface temperatures in Peru. AB - Peruvian sea catfish (Galeichthys peruvianus) sagittal otoliths preserve a record of modern and mid-Holocene sea surface temperatures (SSTs). Oxygen isotope profiles in otoliths excavated from Ostra [6010 +/- 90 years before the present (yr B.P.); 8 degrees 55'S] indicate that summer SSTs were approximately 3 degrees C warmer than those of the present. Siches otoliths (6450 +/- 110 yr B.P.; 4 degrees 40'S) recorded mean annual temperatures approximately 3 degrees to 4 degrees C warmer than were measured under modern conditions. Trophic level and population diversity and equitability data from these faunal assemblages and other Peruvian archaeological sites support the isotope interpretations and suggest that upwelling of the Peru-Chile current intensified after approximately 5000 yr B.P. PMID- 11859191 TI - Abrupt decrease in tropical Pacific sea surface salinity at end of Little Ice Age. AB - A 420-year history of strontium/calcium, uranium/calcium, and oxygen isotope ratios in eight coral cores from the Great Barrier Reef, Australia, indicates that sea surface temperature and salinity were higher in the 18th century than in the 20th century. An abrupt freshening after 1870 occurred simultaneously throughout the southwestern Pacific, coinciding with cooling tropical temperatures. Higher salinities between 1565 and 1870 are best explained by a combination of advection and wind-induced evaporation resulting from a strong latitudinal temperature gradient and intensified circulation. The global Little Ice Age glacial expansion may have been driven, in part, by greater poleward transport of water vapor from the tropical Pacific. PMID- 11859192 TI - Microbial activity at gigapascal pressures. AB - We observed physiological and metabolic activity of Shewanella oneidensis strain MR1 and Escherichia coli strain MG1655 at pressures of 68 to 1680 megapascals (MPa) in diamond anvil cells. We measured biological formate oxidation at high pressures (68 to 1060 MPa). At pressures of 1200 to 1600 MPa, living bacteria resided in fluid inclusions in ice-VI crystals and continued to be viable upon subsequent release to ambient pressures (0.1 MPa). Evidence of microbial viability and activity at these extreme pressures expands by an order of magnitude the range of conditions representing the habitable zone in the solar system. PMID- 11859193 TI - Global allocation rules for patterns of biomass partitioning in seed plants. AB - A general allometric model has been derived to predict intraspecific and interspecific scaling relationships among seed plant leaf, stem, and root biomass. Analysis of a large compendium of standing organ biomass sampled across a broad sampling of taxa inhabiting diverse ecological habitats supports the relations predicted by the model and defines the boundary conditions for above- and below-ground biomass partitioning. These canonical biomass relations are insensitive to phyletic affiliation (conifers versus angiosperms) and variation in averaged local environmental conditions. The model thus identifies and defines the limits that have guided the diversification of seed plant biomass allocation strategies. PMID- 11859194 TI - Enzyme dynamics during catalysis. AB - Internal protein dynamics are intimately connected to enzymatic catalysis. However, enzyme motions linked to substrate turnover remain largely unknown. We have studied dynamics of an enzyme during catalysis at atomic resolution using nuclear magnetic resonance relaxation methods. During catalytic action of the enzyme cyclophilin A, we detect conformational fluctuations of the active site that occur on a time scale of hundreds of microseconds. The rates of conformational dynamics of the enzyme strongly correlate with the microscopic rates of substrate turnover. The present results, together with available structural data, allow a prediction of the reaction trajectory. PMID- 11859195 TI - Effect of p53 status on tumor response to antiangiogenic therapy. AB - The p53 tumor suppressor gene is inactivated in the majority of human cancers. Tumor cells deficient in p53 display a diminished rate of apoptosis under hypoxic conditions, a circumstance that might reduce their reliance on vascular supply, and hence their responsiveness to antiangiogenic therapy. Here, we report that mice bearing tumors derived from p53(-/-) HCT116 human colorectal cancer cells were less responsive to antiangiogenic combination therapy than mice bearing isogenic p53(+/+) tumors. Thus, although antiangiogenic therapy targets genetically stable endothelial cells in the tumor vasculature, genetic alterations that decrease the vascular dependence of tumor cells can influence the therapeutic response of tumors to this therapy. PMID- 11859196 TI - Transcriptional regulation of cortical neuron migration by POU domain factors. AB - The identification of pathways mediated by the kinase Cdk5 and the ligand reelin has provided a conceptual framework for exploring the molecular mechanisms underlying proper lamination of the developing mammalian cerebral cortex. In this report, we identify a component of the regulation of Cdk5-mediated cortical lamination by genetic analysis of the roles of the class III POU domain transcription factors, Brn-1 and Brn-2, expressed during the development of the forebrain and coexpressed in most layer II-V cortical neurons. Brn-1 and Brn-2 appear to critically control the initiation of radial migration, redundantly regulating the cell-autonomous expression of the p35 and p39 regulatory subunits of Cdk5 in migrating cortical neurons, with Brn-1(-/-)/Brn-2(-/-) mice exhibiting cortical inversion. PMID- 11859197 TI - Functional MRI of macaque monkeys performing a cognitive set-shifting task. AB - Functional brain organization of macaque monkeys and humans was directly compared by functional magnetic resonance imaging. Subjects of both species performed a modified Wisconsin Card Sorting Test that required behavioral flexibility in the form of cognitive set shifting. Equivalent visual stimuli and task sequence were used for the two species. We found transient activation related to cognitive set shifting in focal regions of prefrontal cortex in both monkeys and humans. These functional homologs were located in cytoarchitectonically equivalent regions in the posterior part of ventrolateral prefrontal cortex. This comparative imaging provides insights into the evolution of cognition in primates. PMID- 11859198 TI - T cell receptor signaling precedes immunological synapse formation. AB - The area of contact between a T cell and an antigen-presenting cell (APC) is known as the immunological synapse. Although its exact function is unknown, one model suggests that it allows for T cell receptor (TCR) clustering and for sustained signaling in T cells for many hours. Here we demonstrate that TCR mediated tyrosine kinase signaling in naive T cells occurred primarily at the periphery of the synapse and was largely abated before mature immunological synapses had formed. These data suggest that many hours of TCR signaling are not required for T cell activation. These observations challenge current ideas about the role of immunological synapses in T cell activation. PMID- 11859199 TI - Prognostic impact of histologic subtyping of adult renal epithelial neoplasms: an experience of 405 cases. AB - Just two and a half decades ago adult renal cell neoplasms, i.e., those arising from the renal tubules or collecting duct epithelium, were subdivided into two major subtypes: "clear cell carcinoma" and "granular cell carcinoma." Subsequent detailed morphologic and/or cytogenetic studies have resulted in the recognition of several distinctive subtypes of adult renal epithelial neoplasms, which has led to the promulgation of a refined contemporary histologic classification of these tumors. This study examines the prognostic significance of histologic subtyping in accordance with the new classification in a consecutive series of 405 cases treated at a single institution. Cases were histologically classified into 28 (7%) benign tumors [27 (6.7%) renal oncocytomas, 1 (0.2%) metanephric adenoma] and 377 (93%) malignant tumors [255 (63%) conventional (clear cell) renal cell carcinoma, 75 (18.5%) papillary renal cell carcinoma, 24 (5.9%) chromophobe renal cell carcinoma, and 23 (5.7%) renal cell carcinoma, unclassified]. A total of 25 (6.6%) malignant tumors showed evidence of sarcomatoid change. Kaplan-Meier survival analysis with log-rank test showed histologic type (p = 0.002), Fuhrman's nuclear grade (p = 0.001), TNM stage (p = 0.001), vascular invasion (p = 0.001), and necrosis (p = 0.001) to be significantly associated with disease-specific survival and progression-free survival, based on follow-up of 368 patients (mean 64.5 months, median 56 months). The 5-year disease-specific survival for chromophobe renal cell carcinoma, papillary renal cell carcinoma, conventional (clear cell) renal cell carcinoma, and renal cell carcinoma, unclassified was 100%, 86%, 76%, and 24%, respectively; no patient with a benign tumor diagnosis progressed or died of disease. The 5-year progression-free survival for chromophobe renal cell carcinoma, papillary renal cell carcinoma, conventional (clear cell) renal cell carcinoma, and renal cell carcinoma, unclassified was 94%, 88%, 70%, and 18%, respectively. Malignant tumors with sarcomatoid change had a 35% and 27%, 5-year disease-specific and progression-free survival, respectively. Cox proportional hazards regression analysis showed TNM stage (p = 0.001), nuclear grade (p = 0.01), and necrosis (p = 0.05) to be significant predictors of disease-specific survival. In conclusion, our study shows that the histologic categorization of adult renal epithelial neoplasms performed by routine light microscopic hematoxylin and eosin-based examination in accordance with the contemporary classification scheme has prognostic utility. PMID- 11859200 TI - Smooth muscle neoplasms of the urinary bladder: a clinicopathologic comparison of leiomyoma and leiomyosarcoma. AB - We report the clinicopathologic, immunohistochemical, and DNA ploidy findings of 18 leiomyosarcomas of the urinary bladder. In addition, we compare these malignant smooth muscle tumors with 10 cases of urinary bladder leiomyoma. The 14 male and four female patients with leiomyosarcoma ranged in age from 25 to 88 years (mean 64 years). The tumors ranged from 3.0 to 15.0 cm (mean 7.1 cm) in greatest dimension and were moderately to highly cellular, consisting of interlacing fascicles of spindled cells with mild to marked nuclear atypia. Coagulative tumor necrosis was identified in 14 cases (78%), and mitotic activity ranged from 1 to 42 mitotic figures (MF) per 10 high power fields (HPF) (mean 12 MF/10 HPF). Tumors were classified as either high-grade (12 cases) or low-grade (six cases) based on nuclear atypia, mitotic activity, and tumor necrosis. Actin positivity was present in 15 tumors (83%), and desmin immunoreactivity was present in seven tumors (39%). All cases were negative for epithelial markers and S-100. Proliferative activity, as assessed by MIB-1 staining, ranged from 0.1% to 51.4% (median 9.1%). Seven (39%) of the leiomyosarcomas were DNA aneuploid, eight (44%) were tetraploid, and three (17%) were diploid. Five patients underwent radical cystoprostatectomy, one radical cystectomy, seven had partial cystectomy, two underwent pelvic exenteration, and three patients had transurethral resection only. Follow-up information was available on all 18 cases and ranged from 2 to 68 months (mean 22 months). Of the 12 patients with high-grade tumors, six (50%) died of disease from 2 to 20 months (mean 7 months) after diagnosis and three patients (25%) are alive with metastatic tumor. Two of the six patients with low grade leiomyosarcoma died of tumor, 61 and 68 months after diagnosis. There were five male and five female patients with leiomyoma ranging in age from 22 to 78 years (mean 61 years). The tumors ranged from 0.5 to 4.5 cm (mean 1.6 cm) in greatest dimension, were well circumscribed, and had low cellularity. Mitotic activity, necrosis, and cellular atypia were absent, and the tumors were strongly positive for both actin and desmin. MIB-1 staining ranged from 0% to 3.8% (median 0.8%). Seven (87.5%) of the leiomyomas were DNA diploid or near-diploid and one (12.5%) was DNA aneuploid. Six patients were treated with transurethral resection and four with partial cystectomy. All 10 patients were alive at the last follow up (mean follow-up 75 months), and no tumor recurred or metastasized. Our study shows that low-grade leiomyosarcomas are capable of malignant behavior, and high grade leiomyosarcomas appear to behave more aggressively than low-grade tumors. In addition, the diagnosis of urinary bladder leiomyoma should be reserved for noninfiltrative smooth muscle tumors lacking mitotic activity, cytologic atypia, and necrosis. PMID- 11859201 TI - Gastrointestinal glomus tumors: a clinicopathologic, immunohistochemical, and molecular genetic study of 32 cases. AB - Glomus tumors usually occur in the peripheral soft tissues, but similar tumors have also been reported in the stomach and occasionally in the intestines. However, the relationship of these tumors to peripheral glomus tumors and gastrointestinal stromal tumors has not been fully clarified because previous series of gastrointestinal glomus tumors predate availability of immunohistochemistry. This clinicopathologic study examined 32 gastrointestinal glomus tumors. All but one of the tumors were located in the stomach and the remaining tumor was from the cecum. The tumors occurred with a strong female predominance (23 females and 9 males) and a median age of 55 years (range 19-90 years). The gastric tumors typically presented with gastrointestinal bleeding or ulcer-like symptoms, and 14 tumors had mucosal ulceration. Five tumors were incidental findings. The tumor sizes varied from 1.1 to 7 cm (median 2 cm), and most were located in the antrum. Histologically, the tumors typically had a solid pattern of sharply demarcated, round glomus cells with prominent, mildly dilated pericytoma-like vessels. Vascular invasion and focal atypia were relatively common (seen in 11 and 13 cases, respectively), and low mitotic activity (1-4 per 50 high power fields), was seen in 10 cases. Immunohistochemically, all tumors were positive for alpha-smooth muscle actin and calponin, and nearly all had a net-like pericellular laminin and collagen type IV positivity. All tumors were negative for desmin and S-100 protein. Three tumors had focal synaptophysin positivity, but none was positive for chromogranin. All tumors lacked KIT expression and the GIST-specific mutations in the c-kit gene. Follow-up revealed one patient death of metastatic disease to liver at 50 months; this tumor had 1 mitosis per 50 high power fields, but had spindle cell foci, mild atypia, and vascular invasion. Thirteen patients were well and alive after long-term follow up. Gastrointestinal glomus tumors occur almost exclusively in the stomach, and they have a good overall prognosis, but a small, unpredictable potential for malignant behavior exists. These tumors are phenotypically similar to peripheral glomus tumors and differ from epithelioid GISTs. PMID- 11859202 TI - Tissue microarray sampling strategy for prostate cancer biomarker analysis. AB - High-density tissue microarrays (TMA) are useful for profiling protein expression in a large number of samples but their use for clinical biomarker studies may be limited in heterogeneous tumors like prostate cancer. In this study, the optimization and validation of a tumor sampling strategy for a prostate cancer outcomes TMA is performed. Prostate cancer proliferation determined by Ki-67 immunohistochemistry was tested. Ten replicate measurements of proliferation using digital image analysis (CAS200, Bacus Labs, Lombard, IL, USA) were made on 10 regions of prostate cancer from a standard glass slide. Five matching tissue microarray sample cores (0.6 mm diameter) were sampled from each of the 10 regions in the parallel study. A bootstrap resampling analysis was used to statistically simulate all possible permutations of TMA sample number per region or sample. Statistical analysis compared TMA samples with Ki-67 expression in standard pathology immunohistochemistry slides. The optimal sampling for TMA cores was reached at 3 as fewer TMA samples significantly increased Ki-67 variability and a larger number did not significantly improve accuracy. To validate these results, a prostate cancer outcomes tissue microarray containing 10 replicate tumor samples from 88 cases was constructed. Similar to the initial study, 1 to 10 randomly selected cores were used to evaluate the Ki-67 expression for each case, computing the 90th percentile of the expression from all samples used in each model. Using this value, a Cox proportional hazards analysis was performed to determine predictors of time until prostate-specific antigen (PSA) recurrence after radical prostatectomy for clinically localized prostate cancer. Examination of multiple models demonstrated that 4 cores was optimal. Using a model with 4 cores, a Cox regression model demonstrated that Ki-67 expression, preoperative PSA, and surgical margin status predicted time to PSA recurrence with hazard ratios of 1.49 (95% confidence interval [CI] 1.01-2.20, p = 0.047), 2.36 (95% CI 1.15-4.85, p = 0.020), and 9.04 (95% CI 2.42-33.81, p = 0.001), respectively. Models with 3 cores to determine Ki-67 expression were also found to predict outcome. In summary, 3 cores were required to optimally represent Ki 67 expression with respect to the standard tumor slide. Three to 4 cores gave the optimal predictive value in a prostate cancer outcomes array. Sampling strategies with fewer than 3 cores may not accurately represent tumor protein expression. Conversely, more than 4 cores will not add significant information. This prostate cancer outcomes array should be useful in evaluating other putative prostate cancer biomarkers. PMID- 11859203 TI - Primary Ewing's sarcoma/primitive neuroectodermal tumor of the kidney: a clinicopathologic and immunohistochemical analysis of 11 cases. AB - Ewing's sarcoma/primitive neuroectodermal tumor (ES/PNET) is an extraordinarily rare primary tumor in the kidney and can be mistaken for a variety of other round cell tumors, including blastema-predominant Wilms' tumor (WT). Approximately 90% of ES/PNET have a specific t(11;22), which results in a chimeric EWS-FLI-1 protein. Immunohistochemistry for the carboxy-terminus of FLI-1 is sensitive and highly specific for the diagnosis of ES/PNET. WT-1, the WT-associated tumor suppressor gene, is overexpressed in WT but not in ES/PNET. No study has examined FLI-1 or WT-1 expression in renal ES/PNET. The clinicopathologic features of 11 renal ES/PNET were studied along with immunohistochemistry for cytokeratin, desmin, CD99, FLI-1, and WT-1. WT were also immunostained for CD99 (5 cases), FLI 1 (10 cases), and WT-1 (9 cases). The patients (6 men, 5 women) ranged from 18 to 49 years of age (mean, 34 yr). The mean tumor size was 11.8 +/- 3.8 cm (mean +/- standard deviation). Presenting symptoms included abdominal/flank pain and/or hematuria. Grossly, all tumors showed necrosis and hemorrhage, and 4 had cystic change. Microscopically, all tumors showed vaguely lobular growth, primitive round cells, and variable rosette formation. Epithelial, myogenous, or cartilaginous differentiation was not seen. Immunohistochemical results on the renal ES/PNET were cytokeratin (2/8 focal), desmin (0/9), CD99 (8/8), FLI-1 (5/8), and WT-1 (0/8). In comparison, the WT only rarely expressed CD99 (1/5) and did not express FLI-1 (0/10), but were usually WT-1-positive (7/9). Follow-up on 8 cases (mean, 28 mo; range, 6-64 mo) showed 4 lung and pleural metastases, 1 bone metastasis, liver metastasis, 2 local recurrences, and 5 deaths from disease (median time to death, 16.8 mo). No case had distant metastatic disease at presentation. Adjuvant therapy included chemotherapy (8 cases), radiation (3 cases), and bone marrow transplantation (1 case). Our study affirms a unique proclivity of renal ES/PNET for young adults and that it is a highly aggressive neoplasm, with rapid death in many cases, usually after the development of treatment-resistant lung metastases. These tumors must be distinguished from blastema-predominant WT and other primitive renal tumors that require different therapy. FLI-1 and WT-1 immunohistochemistry may be valuable in this differential diagnosis, given the known immunophenotypic overlap between ES/PNET and blastema predominant WT with regard to CD99, cytokeratin, and desmin. The accurate distinction between these two entities has clear prognostic and therapeutic implications. PMID- 11859204 TI - Benign vascular proliferations in irradiated skin. AB - Several types of cutaneous vascular proliferations have been described in areas of irradiated skin, including both benign lesions, such as benign lymphangiomatous papules, atypical vascular lesions, or benign lymphangioendothelioma, and malignant neoplasms such as high-grade angiosarcomas. This report describes the clinicopathologic features of 15 cases of different types of benign cutaneous vascular proliferations arisen within irradiated skin. All patients were female ranging in age from 33 to 72 years, and they had received postoperative external radiotherapy for treatment of breast carcinoma (14 cases) or ovarian carcinoma (one case). In those cases in which the latency interval period between radiotherapy and the development of the vascular lesion was known from the clinical records, the latency interval period elapsed between radiotherapy and diagnosis of the vascular lesion ranged from 3 to 20 years. The most common clinical presentation of the cutaneous lesions consisted of papules, small vesicles, or erythematous plaques on the irradiated field. Histopathologically, most lesions consisted of irregular dilated vascular spaces, with a branching and anastomosing pattern, thin walls, and lymphatic appearance involving the superficial dermis. A discontinuous single layer of endothelial cells with flattened nuclei lined these vascular channels, and numerous small stromal papillary formations also lined by endothelial cells projected into the lumina of the dilated lymphatic vessels. These cases were classified as benign lymphangiomatous papules or plaques. Two cases showed different histopathologic findings because they consisted of poorly circumscribed and focally infiltrating irregular jagged vascular spaces involving the entire dermis and lined by inconspicuous endothelial cells. In some areas these irregular slit-like vascular spaces dissected collagen bundles of the dermis. These cases were classified as atypical vascular proliferations mimicking benign lymphangioendothelioma or patch stage Kaposi's sarcoma. All cases showed similar immunohistochemical findings and the endothelial cells lining the vascular spaces expressed immunoreactivity for CD31, but they stained only focally positive for CD34 or were negative for this marker. Immunohistochemical investigations for alpha-smooth muscle actin failed to demonstrate a complete peripheral ring of actin-positive pericytes in most of the neoformed vascular structures. This immunohistochemical profile also supported the lymphatic nature of these vascular proliferations developed in irradiated skin. Although some of these lesions may mimic histopathologically patch-stage Kaposi's sarcoma or well-differentiated angiosarcoma, the follow-up of the patients of this series demonstrated that the vascular proliferations arisen in irradiated skin invariably showed a benign biologic behavior. PMID- 11859205 TI - Loss of DNA mismatch repair proteins in skin tumors from patients with Muir-Torre syndrome and MSH2 or MLH1 germline mutations: establishment of immunohistochemical analysis as a screening test. AB - Muir-Torre syndrome (MTS) is a rare autosomal-dominant disorder characterized by the predisposition to both sebaceous skin tumors (or multiple keratoacanthomas) and internal malignancies. A subtype of MTS is allelic to hereditary nonpolyposis colorectal cancer and is caused by germline mutations in the DNA mismatch repair genes MSH2 or MLH1. In these cases both internal and skin tumors show characteristic microsatellite instability (MSI). The aim of the present study was to determine whether immunohistochemical examination of MSH2 or MLH1 protein expression in MTS-associated skin tumors can be used as a diagnostic screening tool to identify patients with germline mutations in MSH2 or MLH1. In the present study 28 skin lesions from 17 patients (20 sebaceous gland tumors, 4 sebaceous hyperplasias, 3 keratoacanthomas, and 1 squamous cell carcinoma) were tested immunohistochemically with antibodies against MSH2 and MLH1. Eighteen of these tumors were from eight patients with known MSH2 germline mutations, two tumors were from a patient with a germline mutation in MLH1, and eight microsatellite stable sporadic skin tumors served as controls. One sample had to be excluded because of a lack of immunoreactivity. All eight microsatellite stable tumors expressed both DNA repair proteins. In 15 of the tumors from MSH2 germline mutation carriers, loss of MSH2 expression was observed, one tumor showed reduced MSH2 expression, and one tumor displayed positive immunoreactivity to MSH2. Both tumors of the MLH1 germline mutation carrier showed loss of the MLH1 protein. In conclusion, our findings demonstrate that immunohistochemical testing of MTS related skin tumors is a reliable screening method with high predictive value for the diagnosis of the DNA mismatch repair-deficient MTS. PMID- 11859206 TI - Oxyphil parathyroid carcinomas: a clinicopathologic and immunohistochemical study of 10 cases. AB - Oxyphil parathyroid carcinomas are uncommon neoplasms, and the clinicopathologic features of these tumors are largely unknown. We evaluated the clinicopathologic features of oxyphil parathyroid carcinomas and the expression of cytokeratin 14 (CK14), the high-affinity glucose transporter-4 (Glut-4), as well as the cell cycle proteins p27 and Ki67 and compared these with oxyphil parathyroid adenomas and chief cell parathyroid adenomas and carcinomas. Formalin-fixed, paraffin embedded archival tissues from primary (n = 6) and recurrent (n = 4) oxyphil carcinomas were analyzed and compared with chief cell parathyroid carcinomas (n = 12), oxyphil parathyroid adenomas (n = 38), and chief cell parathyroid adenomas (n = 17) by immunohistochemistry for CK14, Glut-4, p27, and Ki67 using the avidin biotin peroxidase system. Patients with primary oxyphil and chief cell carcinoma presented with high levels of serum calcium (n = 15.5 and 13.7 mg/dL, respectively). Approximately half the patients in each group died of disease. The Ki67 labeling index was higher (4.9 vs 1.9) and the p27 index lower (23 vs 66) in primary oxyphil carcinoma compared with primary oxyphil adenomas. CK14 was expressed in most oxyphil adenomas (35 of 38 cases) but not in oxyphil carcinomas (0 of 10 cases). Glut-4 was more commonly expressed in both groups of adenomas compared with carcinomas. These results show that oxyphil parathyroid carcinomas are functional malignancies similar to chief cell carcinomas and are associated with hypercalcemia, recurrence, and death. Expression of CK14 is very different in oxyphil adenomas compared with carcinomas. Although distinction between parathyroid adenomas and carcinomas can only be made by histopathologic and clinical findings, these results suggest that immunostaining for CK14, p27, and Ki67 may provide additional information to help distinguish between difficult cases of parathyroid adenomas and carcinomas. These findings also indicate that the same histopathologic features should be used to diagnose oxyphil and chief cell parathyroid carcinomas. PMID- 11859207 TI - Circumferential margin involvement is still an important predictor of local recurrence in rectal carcinoma: not one millimeter but two millimeters is the limit. AB - Despite improved surgical treatment strategies for rectal cancer, 5-15% of all patients will develop local recurrences. After conservative surgery, circumferential resection margin (CRM) involvement is a strong predictor of local recurrence. The consequences of a positive CRM after total mesorectal excision (TME) have not been evaluated in a large patient population. In a nationwide randomized multicenter trial comparing preoperative radiotherapy and TME versus TME alone for rectal cancer, CRM involvement was determined according to trial protocol. In this study we analyze the criteria by which the CRM needs to be assessed to predict local recurrence for nonirradiated patients (n = 656, median follow-up 35 months). CRM involvement is a strong predictor for local recurrence after TME. A margin of < or = 2 mm is associated with a local recurrence risk of 16% compared with 5.8% in patients with more mesorectal tissue surrounding the tumor (p <0.0001). In addition, patients with margins < or = 1 mm have an increased risk for distant metastases (37.6% vs 12.7%, p <0.0001) as well as shorter survival. The prognostic value of CRM involvement is independent of TNM classification. Accurate determination of CRM in rectal cancer is important for determination of local recurrence risk, which might subsequently be prevented by additional therapy. In contrast to earlier studies, we show that an increased risk is present when margins are < or = 2 mm. PMID- 11859208 TI - Micropapillary component in lung adenocarcinoma: a distinctive histologic feature with possible prognostic significance. AB - Micropapillary carcinoma or a micropapillary carcinoma component has been reported in the ovary, breast, and urinary bladder and is generally thought to have prognostic significance. However, little has been written on micropapillary differentiation in lung carcinoma. We studied 35 cases of primary lung adenocarcinoma with a micropapillary component seen at the M.D. Anderson Cancer Center. The micropapillary component in these tumors ranged from focal to prominent and was seen at both primary and metastatic sites. This component was not associated with any particular histologic subtype of lung adenocarcinoma. Of the 15 cases with available material, 14 (93%) stained positive for cytokeratin 7, whereas only two of the 15 cases (13%) stained positive for cytokeratin 20. Thyroid transcription factor-1 immunostaining of tumor nuclei was seen in 12 of the 15 cases (80%). Immunostaining was seen in areas both with and without micropapillary differentiation. Thirty-three of 35 patients (94%) developed metastases, which occurred most commonly in the lymph nodes (n = 26), and also in the lung (n = 17), brain (n = 9 cases), bone (n = 9 cases), and other sites. Most metastases had a prominent micropapillary component, irrespective of the extent of the micropapillary carcinoma component in the primary lung tumor. Adequate clinical follow-up information was available for 29 patients. The mean follow-up was 25 months. At their last follow-up, 16 of 29 patients (55%) were still alive with disease, 5 (17%) were dead of disease, and 8 (28%) were alive with no evidence of disease. We believe that a micropapillary component occurring in lung adenocarcinoma should be reported, as this component may be more likely to metastasize. The presence of this component should alert the clinician to search more carefully for metastases and have a closer follow-up on these patients. It is also important to recognize this component in evaluating a metastasis from an unknown primary site, as it should alert the pathologist to a possible primary in the lung in addition to breast, urinary bladder, and ovary. PMID- 11859209 TI - Expression of telomerase reverse transcriptase (TERT) in malignant mesotheliomas. AB - To evaluate the usefulness of determinations of telomerase activity for distinguishing malignant from benign mesothelial lesions, immunohistochemical (using a rabbit polyclonal antibody and the peroxidase method; n = 68) and in situ hybridization (using sense and antisense oligonucleotide probes; n = 46) studies were made on malignant mesotheliomas (epithelioid, 39; sarcomatoid, 18, including 2 of the desmoplastic type; and biphasic, 11) and 19 benign mesothelial lesions (benign mesothelial hyperplasia, 3; and reactive pleuritis, 16). In addition, biochemical studies of telomerase activity were made in 9 of the malignant mesotheliomas. Telomerase activity was detected histochemically in all but one of the malignant mesotheliomas, but only in one (pleuritis) of the benign lesions, in which it was present only in activated lymphocytes. Antisense hybridization signals indicated the presence of telomerase mRNA mainly in the cytoplasm of the malignant cells. Sense probes gave negative results. Biochemical determinations revealed a strong telomerase activity in the 9 malignant mesotheliomas examined. This study demonstrates the usefulness of immunohistochemical staining for the evaluation of mesotheliomas. The required immunostaining can be performed using paraffin sections of formalin-fixed tissues. PMID- 11859210 TI - Sinonasal undifferentiated carcinoma and nasopharyngeal-type undifferentiated carcinoma: two clinically, biologically, and histopathologically distinct entities. AB - Sinonasal undifferentiated carcinoma (SNUC) is a rare aggressive neoplasm arising in the nasal cavity and paranasal sinuses. Primary sinonasal nasopharyngeal-type undifferentiated carcinoma (PSNPC) is an even rarer tumor that has not been adequately reported. Both tumors have been reported to be associated with Epstein Barr virus (EBV). We studied the clinicopathologic features and EBV status of 36 SNUC and 13 PSNPC patients from Taiwan, an EBV endemic area. The median age of SNUC patients was 53 years (range 20-76 years), with a male/female ratio of approximately 2:1. Five patients had histories of previous nasopharyngeal carcinoma treated with irradiation 6-26 years earlier. The most common locations were nasal cavity and ethmoid sinus. Orbital and intracranial invasion and distant metastasis were frequent findings. The median survival was 10 months. All 36 tumors were negative for EBER-1 by in situ hybridization. The median age of PSNPC patients was 58 years (range 36-75 years), with a male/female ratio of approximately 2:1. The most common location is nasal cavity. Eight patients achieved disease-free survival. Eight tumors had the morphology of lymphoepithelioma, whereas significant inflammatory infiltrate was not detected in the other five tumors. All 13 tumors were positive for EBER-1 by in situ hybridization. Because of the difference in the relation with EBV, prognosis, and response to radiotherapy, SNUC and PSNPC should be considered as two entirely different entities. The most important criteria for PSNPC are vesicular nuclei, syncytial pattern, spindle cells, and absence of necrosis. PMID- 11859211 TI - Recommendations for sentinel lymph node processing in breast cancer. AB - The status of the sentinel lymph node (SLN) has been shown to accurately reflect the presence or absence of metastases in the axilla in patients with breast cancer. This study was designed to determine the optimal protocol for SLN processing. A total of 173 SLNs from 96 breast cancer patients who had successful SLN localization and underwent completion axillary node dissection were identified. All SLNs were negative for metastases by initial routine histologic evaluation. The nodes were submitted in a total of 300 blocks. Each block was serially sectioned to produce 10 levels. Pan-cytokeratin stain was performed on levels 3 and 8. All other levels were stained with hematoxylin and eosin. Metastases were identified in 22 SLNs from 19 patients by examining all 10 levels. The first two hematoxylin and eosin- or the first cytokeratin-stained levels were positive for metastases in 21 (95.5%) of the 22 positive SLNs. Two additional hematoxylin and eosin-stained and one cytokeratin-stained levels of each SLN correctly identified the status of the node in 94 (97.9%) of 96 patients. Therefore, we recommend that after an initial hematoxylin and eosin stained section, two additional hematoxylin and eosin-stained sections and one cytokeratin-stained section should be evaluated. PMID- 11859212 TI - Sentinel node processing: recommendations for pathologists. PMID- 11859213 TI - Clear cell chondrosarcoma of the larynx: a case report of a rare histologic variant in an uncommon localization. AB - The authors describe a clear cell chondrosarcoma of the larynx. The clear cell type is a rare variant of chondrosarcoma that only twice has been reported in this localization. The light-microscopic diagnosis of the actual case was confirmed by immunohistochemical results, in particular by positive staining for S-100 protein and collagen type II, and ultrastructural findings. Loss of heterozygosity analysis demonstrated allelic loss at 9p22 and 18q21, but neither in the region of the Rb gene on chromosome 13q nor at the p53 locus on chromosome 17p where allelic loss has already been reported in chondrosarcomas. Furthermore, our molecular genetic investigations revealed a methylation of the cell cycle control gene p16, which is localized on chromosome 9p. This characteristic has been recorded previously only in high-grade chondrosarcomas. Mutations in the exons of p16, alterations of the putative tumor suppressor gene MMAC1/PTEN on chromosome 10q, or an amplification of the cyclin D1 gene (CCND1) on 11q13, which were found to be changed in other studies of chondrosarcomas, could not be demonstrated here. PMID- 11859214 TI - Carney complex. PMID- 11859215 TI - Editorial comment. Volunteerism. PMID- 11859216 TI - The reconstruction of the hand in leprosy. 1952. PMID- 11859218 TI - Orthopaedics in the Buddhist kingdom of Bhutan. AB - Volunteering as the only orthopaedic surgeon in the small Asian country of Bhutan provided a fascinating introduction to international work. The orthopaedic work predominantly was trauma and osteomyelitis. Treatment of most problems was complicated by patient delays in obtaining medical care, usually related to transportation difficulties in a steep mountainous land with few roads. The complexities of enhancing orthopaedic care in developing countries include patient access to care, availability of adequate sterility for implantation of fixation devices, and determining in concert with the local government what is a sustainable level of care that can be taught to interested physicians in the country. Among the developing nations, Bhutan is one with a strong interest in the welfare of its people and has an economy that can afford to invest in healthcare. Despite these strengths, there remain interesting challenges for the orthopaedic volunteer. Exposure of the volunteer to the people and their culture enhanced the experience and provides a lure to continue volunteering. PMID- 11859217 TI - The University of California at San Francisco international orthopaedic elective. AB - International volunteerism helps remedy global inequities in orthopaedic care and provides relief for increasing professional disillusionment experienced by many orthopaedic surgeons in the United States. From 1992 to 1998, 41% of residents from the Department of Orthopaedic Surgery at the University of California, San Francisco volunteered overseas. Approximately one half of those have continued volunteering internationally after residency, including many who led later trips with residents. Based on the success of these trips, the University of California, San Francisco Department of Orthopaedic Surgery established a 1-month elective rotation in Umtata, South Africa in conjunction with Orthopaedics Overseas. Seventy-six percent of residents have chosen this opportunity since the program's inception in 1998. The University of California, San Francisco experience suggests that early exposure to international volunteerism during residency promotes continued participation in volunteer activities after graduation. By providing residents with the opportunity to volunteer overseas, the University of California, San Francisco hopes to enhance resident education, foster a lifelong spirit of volunteerism, and serve as a model for other orthopaedic training programs. PMID- 11859219 TI - The history of orthopaedics overseas. AB - Orthopaedics Overseas is a volunteer organization whose roots began approximately 40 years ago. It started with the Orthopaedic Letters Club with its leader Dr. Allan McElvie and blossomed with the help of Medical International Cooperative Organization (MEDICO), Cooperative for American Relief Everywhere (CARE), and World Orthopaedic Concern (WOC). The organization's goal was to teach orthopaedics in third world countries and has grown to include programs in approximately 16 countries today. This history explains how the organization began and describes some of its great leaders. It also discusses some of the experiences of the volunteers. PMID- 11859220 TI - Volunteerism on the Navajo reservation: the Chinle hand service. AB - In 1994, a pro bono hand clinic was started on the Navajo Nation in northeastern Arizona. The Navajo Nation, larger than the state of West Virginia, holds the greatest concentration of Native Americans in the United States. There has been no specialty hand care on the reservation, home to a population approximately 160,000 people. The hand clinic has operated monthly for the past 7 years with essentially no loss of continuity. More than 2500 patients have been treated. This is an outreach clinic about which the participants can be very proud. PMID- 11859221 TI - Volunteering in Africa: an orthopaedic surgeon-pilot flies to Africa to volunteer. AB - I lived a dream. After 45 years service as an orthopaedic surgeon, a career military officer, private practice, and an appointment to the faculty at Duke University, I retired to begin another career--volunteerism. Specifically, I volunteered to serve at Tenwek Hospital in Africa. Tenwek, a World Medical Mission Hospital, is a 4-hour drive west of Nairobi, Kenya. The hospital serves 400,000 Kenyans. Tenwek is a wonderful hospital, modern by African standards, well-equipped and staffed. The average daily orthopaedic case load is five to nine cases. Fractures, sepsis, and deformity are the usual surgical challenges. An essential part of the dream was to commute to serve-to fly myself across the Atlantic, through Europe, and on to Africa in a manner reminiscent of aviation pioneers long past. This portion of the dream was not realized completely, but was sufficiently achieved to appreciate the accomplishments and difficulties of those who went before. For me, my dream was realized. Volunteer service brings heartache and joy but adds a special dimension to our profession. The motto "We make a living by what we earn, we make a life by what we give" was reinforced each day. PMID- 11859222 TI - Experiences as sports coordinator for the Philadelphia County Special Olympics. AB - Special Olympics is a year-round training and competition program for mentally challenged athletes older than 8 years. Opportunities for involvement in this program are available to members of the orthopaedic community. Participation requires a physical examination every 3 years. The experience with coordinating a sports screening program for 5000 athletes is described. PMID- 11859223 TI - A hand surgeon and his family in Phnom Penh, Cambodia. AB - Those of us who have trained and practice our profession in developed countries, frequently overlook the orthopaedic, and general medical needs, of the developing world. After brief periods in India and more than 4 years practicing in Cambodia, the opportunities for the orthopaedic surgeon to make an impact on patient care and medical education are clear. The challenge of treating patients with untreated congenital and traumatic deformity, advanced tumors, and land mine injuries can be met with dedication to medical education and skills transfer to local personnel. I have experienced many challenges, balanced by the satisfaction of teaching a generation of surgeons and directly helping so many who would have no other opportunity for care, while providing a worthwhile experience personally and for my family. Many will find such work rewarding, knowing that they will leave their mark for good on a hurting world. PMID- 11859224 TI - Physicians and surgeons volunteering in developing countries: a personal perspective. AB - After a physician volunteers in the developing world, his or her eyes open to vast problems. There are many opportunities for physicians to volunteer in developing countries. Medical personnel should know the major agencies that send volunteers and understand how their actions help in disaster relief, direct patient care, or teaching. Volunteers should work with major established organizations to ensure they are not more of a burden than a help. Individuals assisting in training, development, or human rights investigations can change the world by their efforts. PMID- 11859225 TI - Personal perception on mentoring. AB - The current authors discuss mentoring as an important volunteer activity. The roles and responsibilities of the mentor and the protege are described. The mentor's role consists of teaching, coaching, supporting, counseling, and sharing information with the protege. The protege responsibilities are to understand the value of the mentor and seek his or her advice. The importance of mentoring underrepresented minorities, potential and present medical students in orthopaedic surgery is specifically described. The personal experience of one author is given as an example of what can be accomplished and why the current authors think mentoring is important. PMID- 11859226 TI - Volunteerism and Shriners Hospitals for Children. AB - Shriners Hospitals for Children exemplifies the spirit of volunteerism and the remarkable effectiveness of this spirit in providing orthopaedic and burn care to thousands of children around the world. The voluntary efforts of Shriners have included fund raising, hospital planning and development, governance, identification of patients in need, and patient transportation. Until the 1980s, orthopaedic surgeons received little or no remuneration for their work in caring for children in Shriners hospitals. Subsequently, numerous factors have caused a conversion of a part-time voluntary medical staff to full-time and contracted part-time staff. Even so, many Shriners hospitals' medical staff members continue to volunteer their own time in caring for children in less fortunate countries, and as a corporation, Shriners Hospitals for Children has provided volunteer medical teams during times of major international disasters. PMID- 11859227 TI - Volunteer experience overseas. AB - Volunteer experience gained in separate 1-month rotations in three African countries is outlined. Each rotation was unique yet each shared much in common. The latter two rotations were sponsored by Orthopaedics Overseas and as such were more structured than the Liberia experience. Various patients were seen including those with congenital, tumor, traumatic, and developmental conditions. The opportunity to have a cultural exchange at the patient and peer level and to provide a needed service made volunteering a professionally rewarding experience. PMID- 11859228 TI - Evolving volunteerism. AB - The process of volunteering to give orthopaedic care and teaching in developing countries often is evolutionary. Initial expectations of the type of surgery and the response of the patients and the local medical community often are different from what is anticipated. Because of many local factors, such as poorly-funded medical facilities, turnover in personnel, and limited time to accomplish the objectives, achieving long-term results often is frustrating. In an attempt to address the needs of disabled children in developing countries, a network of teaching hospitals for the care of disabled children was begun. A nonprofit organization was formed to raise funds for construction and ongoing expenses to support these hospitals in Kenya, Uganda, Malawi, and the Dominican Republic. Data are being accumulated in an attempt to define the appropriate and most fiscally responsible forms of care for common orthopaedic procedures. A blend of full-time orthopaedic surgeons who are supported by additional short-term volunteers enables a comprehensive teaching program, which ultimately can create major positive changes in the quality of care given to children in the developing world. PMID- 11859229 TI - Personal experiences with overseas volunteerism. AB - Orthopaedic surgeons often are unaware of the many opportunities and rewards of practicing and teaching as an overseas volunteer in a developing country. Opportunities include participating as a member of an American team under the auspices of groups such as Operation Rainbow; or, one can go alone and practice just with host country personnel through organizations such as Orthopaedics Overseas. Typically, the group missions are short-term assignments of 1 to 2 weeks and are more oriented toward doing surgery. The solo visits typically are 1 month or more and aimed more at teaching. The visits are as rewarding and educational to the visiting volunteer as they are to the host country. The volunteer must be open minded and willing to adapt frequently; he or she will be impressed by the ingenuity and resourcefulness of the host orthopaedists who generally work hard with limited equipment and basic supplies. In general, patients are appreciative and rather stoic. Surgeons from the host country also are grateful and eager to learn and share knowledge. PMID- 11859230 TI - Orthopaedics in Indian country. AB - The practice of medicine in rural Arizona has changed remarkably in 25 years. Although the delivery of care to the people of the San Carlos Apache Tribe has many challenges including geographic isolation and cultural differences, there are many rewards, such as knowing that some people would not receive care if the orthopaedic surgeon was not there to provide it. PMID- 11859231 TI - Biomechanical evaluation of the elbow using roentgen stereophotogrammetric analysis. AB - The medial collateral ligament complex is the primary constraint of the elbow to valgus forces and is composed of the anterior bundle, the posterior bundle, and a transverse part. Total and partial ruptures have been described. Clinical and radiologic examinations of medial or valgus instability of the elbow are difficult. The effect of different stages of medial collateral ligament ruptures on ulnohumeral movement in cadavers was determined to rationalize the use of physical and radiologic examinations in different stages of valgus instability in vivo. Using roentgen stereophotogrammetric analysis, motion is determined between the humerus and ulna under valgus load and between the humerus and radius during maximal pronation of the forearm after various dimensions of medial collateral ligament lesions. The increase in distance between the humerus and ulna under a 15 N valgus load varied from 2.7 mm to 9.8 mm. The increase in distance between the humerus and proximal radius with the forearm in pronation in an intact specimen and after transsection of the anterior medial collateral ligament and posterior medial collateral ligament in the anterior direction was 9.7 mm. These results suggest that detection of partial ruptures with clinical and radiologic examinations is difficult. Anterior movement of the radial head can be used as an additional parameter of valgus instability. PMID- 11859232 TI - Perioperative ulnar neuropathy in orthopaedics: association with tilting the patient. AB - The incidence and causes of perioperative ulnar neuropathy in a prospective series of 203 consecutive patients were examined. Three percent of patients had ulnar neuropathy develop in the entire prospective series. The incidence was 6% in patients having total hip arthroplasty. There was a highly significant association between a tilted body position on the operating table and development of ulnar neuropathy on the contralateral side. This position rotates the arm internally and places the ulnar nerve at risk for direct compression. PMID- 11859233 TI - Cubonavicular coalition: a case report and literature review. AB - Tarsal coalitions are uncommon entities. Talocalcaneal and calcaneonavicular synostosis are present in most of the reported cases. Cubonavicular coalition represents only 1% of all the cases in the literature reviewed. The current authors report on a new case of bilateral cubonavicular coalition and present a literature review. The authors also report on the etiology, clinical and radiologic findings, and the treatment to be used. Tarsal coalitions are uncommon entities that must be suspected in an adolescent with painful feet who also has decreased mobility and deformity of the feet. PMID- 11859234 TI - Surface-active phospholipid in total hip arthroplasty. AB - This is the first report of surface-active phospholipid as the boundary lubricant in total hip arthroplasty. Aspirate and rinsings from the bearing surfaces of 25 revision operations and one primary surgery were analyzed from 3 weeks to 26 years postoperatively. All samples contained substantial amounts of surface active phospholipids ranging from 14 to 4186 microg. These findings indicate that synoviocytes continue producing the lubricant in significant quantities after arthroplasty surgery independent of the type of joint replacement and its fixation. Surface-active phospholipid was found on all bearing surfaces analyzed including polyethylene, stainless steel, chrome cobalt, alumina, zirconia, and titanium. PMID- 11859235 TI - Intraoperative autotransfusion in hip arthroplasty. AB - Two groups of patients having primary or revision total hip replacement were studied during a period of 11 months. The first group of 40 consecutive patients consented to have whole blood collected coincident with acute normovolemic hemodilution. An inventory of autologous red blood cells, plasma, and platelets was prepared in the operating room as a preliminary to surgery. Subsequently, the same supplies and equipment were used for autotransfusion intraoperatively. In a case-control study, another 40 patients having total hip replacements were matched for age, gender, and weight. The second group of patients refused hemodilution and received autotransfusion alone. The perioperative transfusion requirements of the two groups were compared. The total blood product donor exposure rate of the first group was 1/4 of the controls (0.6 and 2.4 donor units per patient, respectively). The average length of stay in the hospital after surgery for the first group was shortened significantly (6.2 versus 8.4 days), possibly from less immunogenic insult associated with increased transfusions of autologous blood products. Hemodilution, followed by autotransfusion, was cost effective in primary and revision total hip replacements, autotransfusion alone was cost effective only in revision arthroplasty. PMID- 11859236 TI - Incorporation of morselized bone grafts: a study of 24 acetabular biopsy specimens. AB - Animal studies have shown almost complete incorporation of impacted morselized bone grafts. To determine whether this also is true in humans, 24 acetabular bone biopsy specimens from 21 hips of 20 patients were examined. Biopsy specimens were obtained 3 months to 15 years after acetabular reconstruction in primary and revision total hip arthroplasties in combination with a cemented cup. Histologic examination showed rapid revascularization of the graft, directly followed by osteoclastic resorption and woven bone formation on the graft remnants. New bone also was formed on fibrin accumulations or without any scaffold in the fibrous stroma tissue that had invaded the graft. Thereafter the mixture of graft, new bone, and fibrin was remodeled completely into a new trabecular structure, with normal lamellar bone and only scarce remnants of graft material. Localized areas of nonincorporated bone graft surrounded by fibrous tissue remained, irrespective of the followup period. Large nonincorporated fragments of cartilage also were found, particularly in cases in which femoral head bone chips were produced by a bone mill. In general, impacted trabecular bone chips incorporate by a mechanism that is similar to that previously observed in animal studies. In some patients, however, areas of nonincorporated bone graft remained and long-term alterations were found, probably related to the loosening process. PMID- 11859237 TI - Intraoperative monitoring for safety of bilateral total knee replacement. AB - Cardiopulmonary hemodynamics in 79 consecutive patients who had one-stage bilateral total knee replacements were monitored prospectively with a pulmonary artery catheter. The pulmonary vascular resistance, wedge pressure, pulmonary artery pressure, and systemic vascular resistance were measured before skin incision, 5 to 10 minutes after implanting the first total knee replacement, and after the second knee replacement. The second knee replacement was cancelled in five patients because the pulmonary vascular resistance after the first knee replacement was more than double the baseline, or above 200 dyne/second/cm(5). No patient had clinical symptoms of fat embolism during the postoperative course. Patient predictive factors, or the use of pulse oximetry readings instead of a pulmonary artery catheter, were not predictive of intraoperative elevation of pulmonary vascular resistance. For this reason, the safety of this operation for the patient requires that intraoperative measurement of hemodynamic parameters of embolism be done. PMID- 11859238 TI - Functional outcome after tibial tubercle transfer for the painful patella alta. AB - Twenty-five patients with painful patella alta without symptomatic subluxation were identified in a prospective database. All patients had a distal tibial tubercle transfer and preoperative knee arthroscopy. The mean postoperative followup was 2.4 years. These patients were matched with healthy volunteers. Patellofemoral scores using the scoring systems of Kujala et al and Lysholm and Gillquist were collected prospectively. The Short Form-36 health survey and the Western Ontario and McMaster Universities Osteoarthritis Index were used postoperatively. Significant improvement in the patellofemoral scores was documented postoperatively; however, the healthy volunteers had significantly higher patellofemoral scores when compared with the patients who were treated surgically. For the three Short Form-36 survey parameters based on physical health (physical functioning, role physical, and bodily pain), there were no statistically significant differences between the patients and the United States age-matched norms; data are available in the Short Form-36 survey manual. Patients with Grade 2 chondromalacia (fissuring and fragmentation less than 1.25 cm) had significantly better scores in pain and function domains of the Western Ontario and McMaster Universities Osteoarthritis Index compared with patients with Grade 3 (fissuring and fragmentation greater than 1.25 cm) and Grade 4 (erosion down to bone) changes. Distal tibial tubercle transfer is a beneficial procedure for treating patients with painful patella alta. PMID- 11859239 TI - Interfragmentary motion in tibial osteotomies stabilized with ring fixators. AB - Relative movement of bone fragments affects healing processes. In vivo data exist for patients with reduced transverse fractures only. The gap movements that occur under more complex conditions such as in tibial osteotomies, however, are unknown. The goal of this study was to determine the initial gap movements in tibial correction osteotomies, to monitor movements during the early healing period, and to determine the suitability of initial fixation stability in relation to daily activities. The interfragmentary movements were measured in six patients with correction osteotomies stabilized by Ilizarov ring fixator constructs until union or until conversion to internal fixation. Consolidation was determined by clinical evaluation and standard radiographic techniques. Co contraction led to gap movements comparable with level walking or standing. Shear generally exceeded axial compression. Although ground reactions and fixation stiffness were comparable with those reported for reduced fractures, movement magnitudes generally were larger than 2 mm. The shear movement component reflected the nature of the mechanical conditions at the bone gap. In a direct comparison with data from animal experiments, the local mechanical environment at the bone gap seemed unstable rather than overly stable. The method introduced in this study opens the perspective of adjusting osteosynthesis stability to the specific needs of each patient. PMID- 11859240 TI - Proximal dome-shaped osteotomy for symptomatic bunionette. AB - The results of a dome-shaped osteotomy of the proximal third of the fifth metatarsal in patients with symptomatic bunionette deformity were reviewed. The series was comprised of eight patients (10 feet; mean age of patients, 21 years). The average followup was 30 months. All patients were free from pain at the fifth metatarsophalangeal joint and were satisfied with the results of this procedure. The mean angle between the longitudinal axes of the fifth metatarsal and the proximal phalanx was 18.9 degrees before surgery and 2.6 degrees after surgery. The mean angle between the longitudinal axes of the fourth and fifth metatarsals was 12.2 degrees before surgery and 4.8 degrees after surgery. The overall results were good in all 10 feet. Three feet had delayed union at the osteotomy site, but union was obtained in all feet. The osteotomy site of the fifth metatarsal in feet with delayed union was more proximal than that of the other feet. Therefore, proximal osteotomy of the fifth metatarsal should be done not at the base, but at the proximal site of the diaphysis to prevent delayed union. A proximal dome-shaped osteotomy corrects the deformity and relieves the symptoms, but careful attention should be paid to the osteotomy site. PMID- 11859241 TI - Adult hallux valgus with metatarsus adductus: a case report. AB - It is difficult to surgically treat a hallux valgus deformity with significant metatarsus adductus, because the space between the first and second metatarsals is too narrow to correct the metatarsus primus varus with a first metatarsal osteotomy. A 55-year-old woman had severe hallux valgus with significant adduction of the second and third metatarsals. A distal soft tissue procedure and a proximal crescent-shaped osteotomy of the first metatarsal combined with corrective osteotomies of the second and third metatarsals were done. The patient's symptoms disappeared, and hallux valgus and adduction of the second and third metatarsals were corrected. To the authors' knowledge, there is no previous description of surgical treatment including correction of metatarsus adductus and hallux valgus for adult hallux valgus with metatarsus adductus. PMID- 11859242 TI - Expression of CD44 isoforms correlates with the metastatic potential of osteosarcoma. AB - CD44 is a family of cell surface glycoproteins that not only function as receptors to hyaluronan and other extracellular matrix ligands but also is implicated in the invasiveness and metastasis of malignancies. The expression of CD44 and its variant isoforms was evaluated by immunohistochemistry in primary osteosarcoma, and the metastatic significance and prognostic relevance were assessed. Fifty formalin-fixed, paraffin-embedded conventional osteosarcoma samples were immunostained with monoclonal antibodies to CD44s, CD44v5, and CD44v6. When more than 50% of the tumor cells were stained, the result was regarded as ++ (overexpression). Staining patterns of CD44 mainly were membranous with a minor cytoplasmic pattern contribution. CD44s was overexpressed in 16% (eight of 50), CD44v6 in 20% (10 of 50), and CD44v5 in 46% (23 of 50) of the osteosarcomas. Of 26 patients with metastasis, CD44s was overexpressed in six (23%), CD44v6 in four (15%), and CD44v5 in 17 (65%). Overexpression of CD44v5 correlated significantly with metastasis. The survival rate of the patients in the CD44v5-positive (overexpressed) group was significantly lower than that of the patients in the CD44v5-negative group. Overexpression of CD44v5 may play a role in the metastatic behavior of osteosarcoma and may be a prognostic parameter of osteosarcoma. PMID- 11859243 TI - Surgical treatment of bone metastases in patients with breast cancer. AB - In this retrospective study, the effect of surgical therapy on a series of 70 patients with breast cancer who were surgically treated for metastasis of the bone was evaluated. At presentation, 19 patients had one osseous lesion, 19 patients had multiple bone lesions, and 32 patients had additional visceral involvement. The surgical procedures included 60 palliative procedures, six radical resections, and four biopsies. In 14 surviving patients, the mean observation period was 35.6 +/- 40.1 months. Of the six patients with radically resected solitary bone lesions, five patients had systemic progression of the disease develop. Of the 19 patients with presumably solitary bone lesions, five currently are free of tumor. Of the 19 patients with multiple bone lesions and initially no visceral tumor spread, only two are alive. Of the 32 patients with additional visceral metastases at surgery, four are alive with the disease. For the entire group, the survival rate was 59% after 1 year, 36% after 2 years, 13% after 5 years, and 7% after 10 years. The only two independent factors that were associated with survival were the extent of the disease and the duration of symptoms from bone metastasis. These findings suggest that in orthopaedic surgery in patients with bone metastases secondary to breast cancer, wide resection is not likely to be necessary. Patients with solitary bone lesions have a 39% chance of living 5 years. PMID- 11859244 TI - Implantation of hemipelvic prosthesis after resection of sarcoma. AB - Twelve adult patients with pelvic sarcoma had implantation of a hemipelvic prosthesis. Eight patients had hemipelvic resection, and four patients had acetabulopubic resection. The implanted prosthesis was a special Vitallium prosthesis, which was specially designed for each patient with the aid of a computer. At a median followup of 57 months, eight patients were free of disease. In four patients with local relapse, two had additional resection, one had hindquarter amputation, and one was observed. In three patients with deep infection, the prosthesis was removed; however, one patient had hindquarter amputation. One patient had dislocation of the hip and prosthesis loosening. Overall survival of patients was 70%, and the survival of prostheses was 42%. In eight patients, the functional evaluation showed that the average functional score with the prosthesis was 11.6 (39%) and without the prosthesis the functional score was 7.0 (23%). Implantation of a megaprosthesis seems to indicate a high complication rate and a poor functional result after hemipelvic resection. PMID- 11859245 TI - Metal-associated angiosarcoma of bone: report of two cases and review of the literature. AB - Angiosarcoma is an extremely rare bone tumor. The authors report two cases of patients with angiosarcoma that developed adjacent to a stainless steel plate used for fixation of a femur fracture. In both patients, the interval between fracture treatment and the development of the neoplasm was more than 40 years. A review of the literature found 36 previously reported cases of malignancy arising adjacent to an orthopaedic implant. Despite any evidence directly linking these implants to the development of cancer, continued vigilance is warranted. PMID- 11859246 TI - Bone grafting of cryosurgically treated bone defects: experiments in goats. AB - It is hypothesized that cryosurgically treated bone defects are inappropriate host sites for cancellous bone grafting. The influence of autologous cancellous bone grafting on the healing of cryosurgically treated gap defects of long bones was investigated. A unilateral in vivo experiment was done to study bone strength and graft incorporation in the goat. The lining of a cylindrical defect of the femoral diaphysis was treated with a closed liquid nitrogen cryoprobe in 62 goats. Thirty-one animals received an impacted, morselized, cancellous bone graft harvested from the sternum. The other 31 animals served as controls. At 0, 4, 7, 10, 13, 16, and 26 weeks animals were euthanized and the femurs were evaluated for torsional strength, computed tomography, and histologic assessment. Specimens with a bone graft showed no significant increase in torsional strength with time compared to the controls. In all goats euthanized at 10 weeks or later, the graft was resorbed. The amount of bone apposition at the site of the cryosurgical lesion and the time at which the defect was bridged were similar in both groups. Autologous cancellous bone grafting does not accelerate healing of cryosurgically treated, stable, diaphyseal defects in the goat. PMID- 11859247 TI - Effect of synergistic motion on flexor digitorum profundus tendon excursion. AB - Clinical and experimental studies have shown that postoperative rehabilitation is an important factor that improves digital function after flexor tendon repair. In the current study, the effect of synergistic wrist and finger motion therapy and fixed flexed wrist motion therapy on the in vivo gliding excursion was evaluated after repair of partial laceration of the canine flexor digitorum profundus tendon. The gliding excursion of the repaired tendons treated with synergistic wrist and finger motion therapy was significantly greater than that of tendons rehabilitated with the wrist fixation therapy, suggesting that wrist extension generates force that can pull the repair site through the pulley, thereby increasing passive excursion of the tendon. As a result of increased tendon excursion, synergistic therapy may improve the clinical outcome after repair of partial tendon lacerations. PMID- 11859248 TI - Biomechanical analysis of humeral diaphyseal segmental defect fixation. AB - With disseminated malignancies, segmental defects of the humeral diaphysis may occur from surgical resection or as a complication of failed prior internal fixation. This study directly compared the biomechanical properties of a second generation titanium modular intercalary humeral spacer (segmental defect replacement prosthesis) with those of a modern locked humeral nail combined with methylmethacrylate (intramedullary nail) or with an intercalary allograft spacer (allograft nail composite) for fixation of segmental defects of the humeral diaphysis. Eighteen matched pairs (36 specimens) of fresh-frozen humeri were prepared in a standard fashion to create a 5-cm middiaphyseal defect and were divided randomly into three groups of 12 specimens each, using three different reconstructive methods. Specimens were tested in external torsion to failure on a Materials Testing System machine. The segmental defect replacement specimens had statistically greater peak torque (mean, 41.4 N-m) and stiffness (mean, 2.1 N-m/ degrees) than the intramedullary nail specimens (mean peak torque, 23.1 N-m) (mean stiffness, 1.6 N-m/ degrees) or the allograft nail composite specimens (mean peak torque, 12.4 N-m) (mean stiffness, 0.6 N-m/ degrees). The intramedullary nail specimens also had a statistically greater peak torque at failure and stiffness than the allograft nail composite specimens. For segmental defects of the humeral diaphysis, reconstruction with a cemented metallic intercalary spacer provides significantly greater immediate stability than interlocked intramedullary nail fixation supplemented with segmental methylmethacrylate or intercalary allograft reconstruction. PMID- 11859249 TI - Perforations of cortical bone allografts improve their incorporation. AB - The incorporation of perforated cortical bone allografts was compared with non perforated allografts. A 5-cm circumferential defect in the middiaphysis at the tibia was created in adult sheep. A frozen tibial allograft was implanted and fixed with a locked nail for 6 months. There was no postoperative immobilization. Group I consisted of eight sheep with non-perforated allografts, whereas Group II was comprised of 10 sheep with perforated allografts. Union was evaluated radiographically, whereas the central part of the allograft had a densitometric evaluation. Creeping substitution was assessed on microradiographs from cross sections of the central 3 cm of graft by measurement of porosity and percentage of new and old bone area within the confines of the graft. The width of periosteal and endosteal callus also was determined. There was no statistical difference between both groups for the union score and bone density. However, the cortical bone graft porosity and the amount of new bone within the cortical bone differed significantly between the perforated allografts and the non-perforated ones. Periosteal callus was similar in both groups, whereas endosteal callus was significantly more wide and extended in the perforated allografts. Perforation of a cortical bone substantially improved the amount of newly formed bone by the host when compared with a non-perforated bone. The creation of channels seemed to increase the interface between living soft tissues of the host and the allografted bone with a resulting enhanced incorporation. PMID- 11859250 TI - Decreases in bone blood flow and bone material properties in aging Fischer-344 rats. AB - The purpose of this study was to quantify precisely aging-induced changes in skeletal perfusion and bone mechanical properties in a small rodent model. Blood flow was measured in conscious juvenile (2 months old), adult (6 months old), and aged (24 months old) male Fischer-344 rats using radiolabeled microspheres. There were no significant differences in bone perfusion rate or vascular resistance between juvenile and adult rats. However, blood flow was lower in aged versus adult rats in the forelimb bones, scapulas, and femurs. To test for functional effects of this decline in blood flow, bone mineral density and mechanical properties were measured in rats from these two age groups. Bone mineral density and cross-sectional moment of inertia in femoral and tibial shafts and the femoral neck were significantly larger in the aged versus adult rats, resulting in increased (+14%-53%) breaking strength and stiffness. However, intrinsic material properties at midshaft of the long bones were 12% to 25% lower in the aged rats. Although these data are consistent with a potential link between decreased perfusion and focal alterations in bone remodeling activity related to clinically relevant bone loss, additional studies are required to establish the mechanisms for this putative relationship. PMID- 11859251 TI - Knee pain and swelling in a 5-year-old girl. PMID- 11859252 TI - Renal medicine and renal transplantation. PMID- 11859253 TI - New insights in chronic allograft rejection. AB - Chronic allograft nephropathy remains the main cause of renal graft failure. Immunologic mechanisms seem mostly responsible for the injury and subsequent fibrogenic tissue response while nonimmune mechanisms act mostly as progression factors. In this article, these factors are reviewed along with the changes that take place in the graft and new insights into possible therapeutic strategies. PMID- 11859254 TI - Acute humoral renal allograft rejection. AB - In kidney transplantation, it is well established that donor-specific antibodies can cause substantial graft injury. Hyperacute rejection, now virtually eliminated by routine pretransplant cytotoxic crossmatch testing, represents the prototype of humoral rejection. However, there is now increasing evidence that alloantibody-mediated immune reactions may also cause acute rejection. Acute humoral rejection, which is frequently associated with severe graft dysfunction and immunologic graft loss, represents a particular diagnostic and therapeutic challenge. Reliable detection of antibody-mediated graft injury is required to govern the application of antihumoral therapeutic strategies. This review focuses on new approaches in the diagnosis and treatment of acute humoral rejection. Special attention is given to a novel diagnostic marker, the complement split product C4d. PMID- 11859255 TI - The marginal kidney donor. AB - In parallel with increased clinical demand, less than optimal or so-called marginal kidney grafts are being used for transplantation. In addition to donor age, it appears that several factors may impact negatively on the quality of the graft. Most importantly, a more precise definition of the term 'marginal graft' is needed. The present review analyzes potential risk factors, suggests scoring systems for a more precise definition, and discusses potential treatment options to improve the quality of marginal grafts. PMID- 11859256 TI - The future role of target of rapamycin inhibitors in renal transplantation. AB - Immunosuppressant nephrotoxicity is among the major contributors to chronic renal allograft failure, which is the primary cause of graft loss. Because of a lack of alternatives to the inherently nephrotoxic calcineurin inhibitors for maintenance immunosuppression, long-term survival rates for renal allografts have not increased in proportion to the rise in short-term graft survival. Clinical studies have shown that mammalian target of rapamycin-based immunosuppression in combination with calcineurin inhibitors, mycophenolate mofetil, or azathioprine is safe and efficacious. These data suggest that a target of rapamycin antagonist (sirolimus/everolimus) should be used initially in combination with calcineurin antagonists in order to prevent early acute rejection. After 3-6 months, a maintenance immunosuppressive regimen can then be individually tailored to each patient on the basis of their clinical and histological status. Those patients at high immunological risk should remain on full-dose triple therapy. All other patients should receive either a calcineurin inhibitor or corticosteroid-sparing regimen, with a maintenance dose of a target of rapamycin inhibitor. This regimen should result in less immunosuppressant nephrotoxicity and a reduction in the serious side effects of steroids, such as diabetes and osteoporosis. Whether the proposed individually designed immunosuppressive regimen, based on protocol biopsies and mammalian target of rapamycin inhibition, will result in prolonged graft and patient survival remains to be determined. PMID- 11859259 TI - Imaging: new diagnostic techniques. PMID- 11859257 TI - Opportunistic infections after renal transplantation. AB - Opportunistic infection is a serious clinical complication in patients receiving immunosuppressive therapy after kidney transplantation. This article deals with some of the possible infectious agents that were recently encountered at our transplantation centre in Dusseldorf, Germany. Opportunistic organsims such as human herpesviruses 6-8, polyomavirus, parvovirus B19, varicella zoster virus, Nocardia and Listeria monocytogenes are rare but severe complications that are presented in this overview. As a result of the use of new immunosuppresive drugs like tacrolimus and mycophenolate mofetil these infections are now seen more frequently, so they should always be included in differential diagnostic considerations. New diagnostic procedures and new treatment strategies should allow early detection and successful treatment of opportunistic infections in the majority of kidney transplant recipients. PMID- 11859258 TI - Urinary tract infections after renal transplantation. AB - Up to now one of the major problems for successful organ transplantation has been the reaction of the immune system of the recipient against the donor organ. This could lead to acute and chronic rejection, and in cases of unsuccessful treatment to the loss of the transplant. In organ graft recipients, immunosuppressive agents are used to prevent or treat rejection episodes and to maintain graft function. Although there is an increasing number of immunosuppressive substances, the immunosuppressive therapy currently in use is relatively unspecific and targets many immunological functions. The net state of immunosuppression is a complex function determined by the interaction of a number of factors, the most important of these are the dose, duration and temporal sequence in which immunosuppressive drugs are employed. Any kind of immunosuppressive protocol is thus associated with an increased infection rate. This has an important socioecological impact, because frequent hospitalizations resulting from infectious complications are necessary, having an overall mortality rate of 3.5% within 2 weeks of admission. The most common cause of septicaemia is urinary tract infection. Frequent urinary tract infections are associated with the early onset of chronic rejection, suggesting a pathogenetic relationship between these two features. The occurrence of chronic rejection has led to reduced transplant survival. The prevention of urinary tract infections, or the early diagnosis and accurate treatment of urinary tract infections is important in renal transplant recipients. PMID- 11859260 TI - Magnetic resonance imaging for urinary incontinence. AB - Magnetic resonance imaging provides the most accurate, versatile and safe imaging of the pelvic floor. Images can be produced to show sections in any plane and even in three dimensions. The resolution is such that detail as good as that seen in histological sections is possible. Once standardization of data acquisition and patient positioning is agreed we look forward to a new era of increasingly accurate diagnoses of incontinence, allowing tailored management, both surgical and nonsurgical. PMID- 11859261 TI - Computed tomography urography, three-dimensional computed tomography and virtual endoscopy. AB - Spiral computed tomography technology allows an entire body region to be imaged as a continuous volume of computed tomography data. The acquisition of genitourinary images can be performed at different intervals after intravenous contrast injection in order to characterize the renal vasculature, the renal parenchyma or the collecting system. Computed tomography scanning as contrast is excreted into the collecting system is termed a 'computed tomography urogram'. Volumetric data from spiral computed tomography can be rendered into conventional two-dimensional images or even reformatted into three-dimensional views of organ systems or hollow structures, as in 'fly-through' virtual endoscopy. Although virtual endoscopy of the urinary tract remains in its infancy, three-dimensional imaging is currently a useful adjunct in the evaluation of renal transplant and donor patients and partial nephrectomy candidates. The role of computed tomography urography compared with intravenous urography in the evaluation of hematuria is discussed. PMID- 11859262 TI - Imaging for erectile dysfunction. AB - Radiological imaging in the field of erectile dysfunction has diminished in importance over the past 5 years with the advent of new therapies. However, in selected cases, Doppler scanning continues to have a role, and current research is aimed at optimizing the results of the tests. In addition, the new techniques of functional magnetic resonance imaging and positron emission tomography scanning are beginning to offer new insights into the central mechanisms which control erections. PMID- 11859263 TI - Scrotal imaging. AB - Color Doppler ultrasound has been the mainstay for the evaluation of the scrotum in a variety of clinical settings. However, ultrasonography results are not always accurate or conclusive. Despite the high cost and limited availability, magnetic resonance imaging with the dynamic contrast-enhanced subtraction technique provides accurate information on morphology as well as blood flow. Infrared scrotal thermography increases accuracy in the diagnosis of varicocele. This article attempts to summarize recent advances in scrotal imaging with regard to testicular and extratesticular disorders. PMID- 11859264 TI - Imaging for microscopic haematuria. AB - For evaluation of the upper urinary tract, intravenous urography has for many years been the method of choice. In recent years, different imaging protocols have been suggested. In this article different strategies for the evaluation of microscopic haematuria, with a focus on the advantages and disadvantages of different imaging modalities, will be discussed. PMID- 11859265 TI - Endoscopic data acquisition and storage. AB - Recent developments in the field of digital image acquisition and storage provide new opportunities for recording endoscopic procedures for use in the medical record and remote access for education and consultation. This article reviews the recent literature in the field of endoscopy. PMID- 11859267 TI - Depot-specific differences in the lipolytic effect of leptin on isolated white adipocytes. AB - BACKGROUND: In the present study we hypothesized that leptin-induced lipolytic effects show depot-specific differences. MATERIAL/METHODS: Three-month-old male lean (+/+) and obese fa/fa rats were used. Lipolysis was determined in the absence or presence of leptin (0.63, 6.25 and 62.5 nM) together with agents acting at the A1 adenosine receptor of fat cells isolated from both subcutaneous and omental depots. The glycerol released into the incubation medium was taken as the lipolytic rate index. RESULTS: The highest dose of leptin produced a 57.5 +/- 7.3% and 70.3 +/- 3.8% increase in omental and subcutaneous glycerol release, respectively, compared to the basal lipolytic rate (P<0.001). The addition of the three leptin concentrations in the ligand-free state produced a significantly greater stimulation of lipolysis in subcutaneous fat cells (P=0.0331; P=0.0003 and P=0.0015, respectively) compared to omental adipocytes. Under adenosine A1 receptor agonism and antagonism the same pattern of response was observed between visceral and subcutaneous adipocytes of lean rats. Although adenosine deaminase produced near maximum lipolysis in the adipocytes of lean animals, only half of the maximum lipolytic rate (50.9 +/- 3.2%) was achieved in fat cells from fa/fa rats (P=0.0034). Leptin had no effect on the lipolytic activity of adipocytes from either localization in fa/fa rats. CONCLUSIONS: It can be concluded that decreased sensitivity to the lipolytic effect of leptin in omental adipocytes compared to subcutaneous fat cells may underlie, at least in part, the association of visceral fat accumulation with increased co-morbidities. PMID- 11859268 TI - A prolonged time interval between blood sample collection and centrifugation causes an increase in serum carbohydrate-deficient transferrin. AB - BACKGROUND: Carbohydrate-deficient transferrin (CDT) is used for the laboratory diagnosis of chronic alcohol abuse. Non-optimal preanalysis can cause an increase in CDT and false positive results. The aim of our study was to determine whether CDT results change over time between collection of the blood sample and centrifugation, and whether shipment of whole blood samples is a potential source of false positive CDT reports. MATERIAL/METHODS: 152 blood samples were drawn from 38 persons (4 tubes per person, one venipuncture) and randomly assigned to 4 groups with different time intervals between blood sample collection and centrifugation (1h, 24h, 48h, 144h). CDT analysis was done using the ChronAlcoI.D. assay. The statistical analysis was based on box-plots, ANOVA and Kruskal-Wallis ANOVA. RESULTS: The means and medians of CDT increased with the time of whole blood storage. ANOVA analysis of between-group differences was significant for mean CDT concentrations between 1 and 144 hours of whole blood storage. There was no correlation between CDT and free hemoglobin as a measure of hemolysis. An interference of hemolysis with CDT measurement can be excluded as the main cause of increased CDT results with whole blood storage time. Whether an in vitro degradation of the transferrin N-glycan chains causes the CDT increase should be evaluated by isoelectric focusing of the transferrin isoforms in a further study. CONCLUSIONS: Storage or shipment of whole blood samples can shift initially normal CDT values to borderline and borderline to pathological CDT results. PMID- 11859269 TI - Antiapoptotic effect of (-)-deprenyl in rat kidney after ischemia-reperfusion. AB - BACKGROUND: Since apoptosis of renal tubular cells is the basis of the damage caused by ischaemia-reperfusion, the antiapoptotic effect on kidney tubular epithelial cells of the monoamino oxidase-B (MAO-B) inhibitor (-) deprenyl (selegiline), known neuroprotective agent, with antiapoptotic properties, was studied in a rat model. MATERIAL/METHODS: Warm renal ischaemia was caused by clamping the left renal artery of rats for 30 minutes. With the start of reperfusion 0.015 mg/kg, 0.15 mg/kg and 1.5 mg/kg of (-)-deprenyl was injected simultaneously into the carotid artery of the animals, respectively. Five rats served as control, in which renal artery clamping was performed, but the rats were only treated with the solvent (physiological saline). After 6 hours of reperfusion the rats were exsanguinated and the kidneys were histologically examined. RESULTS: Severe tubular damage characterised by apoptosis was found in the kidneys of the untreated rats. Apoptosis was verified on the basis of morphological features, methylgreen-pyronin staining and TUNEL reaction. (-) Deprenyl diminished dose-dependently the apoptotic damage, 0.15 mg/kg being the most effective dose. The same dose of (-)-Deprenyl is used in the therapy of human Parkinson's disease. CONCLUSIONS: Our findings suggest, that (-)-deprenyl might have an impact on decreasing renal injury also in case of human cadaveric renal transplantation. PMID- 11859270 TI - Influence of losartan and enalapril on urinary excretion of 8-isoprostane in experimental nephrotic syndrome. AB - BACKGROUND: The increased permeability of the glomerular capillary wall in adriamycin nephropathy may be mediated by increased generation of free radicals, possibly also by the non-enzymatic production of isoprostanes induced by oxidative stress. ACE inhibitors and angiotensin II antagonists may reduce proteinuria, perhaps by decreasing intraglomerular pressure and increasing the selective permeabiity of the glomerular capillary wall. MATERIAL/METHODS: We compared the effect of an ACE inhibitor, enalapril, and an angiotensin II antagonist, losartan, on total malodialdehyde in blood and the urinary excretion of certain eicosanoids and their metabolites (TxB(2), 6-keto-PGF(1alpha), bicyclo PGE(2) and 8-isoprostane) in experimental adriamycin-induced nephrotic syndrome in rats. RESULTS: Increased proteinuria in adriamycin-treated rats was not prevented by losartan, but tended to be partly mitigated by enalapril. However, both losartan and enalapril prevented the adriamycin-induced increase of total MDA in serum, but urinary excretion of 8-isoprostane was increased in nephrotic rats treated by losartan compared to controls. The enalapril-induced increase in urinary excretion of bicyclo-PGE(2) was possibly mediated by kinins. Proteinuria positively correlated with urinary excretion of 8-isoprostane, and proteinuric rats also had a significantly higher urinary excretion of 8-isoprostane than non proteinuric rats. CONCLUSIONS: Proteinuria in the acute phase of adriamycine nephropathy may be dependent on free radical generation and the formation of 8 isoprostane. The mild antiproteinuric effect of enalapril, but not losartan, may suggest the contributory role of the inhibition of kinin degradation in the antiproteinuric action of enalapril in this model of nephrotic syndrome. PMID- 11859271 TI - High affinity glutamate uptake in rat brain slices at experimental crush syndrome. AB - BACKGROUND: The pathogenesis of crush syndrome is severe traumatic damage to the organism accompanied by shock and stress, acute toxemia, etc. Glutamate has been shown to be implicated in excitotoxic neural death in various clinical settings. The high affinity glutamate uptake process plays a key role in normal synaptic transmission. The goal of our research was to determine the level of glutamate uptake in brain tissue slices after incorporation of [(14)C-U]glutamate. MATERIAL/METHODS: An experimental model of crush syndrome in Wistar male rats was caused by 2 hours of compression of femoral soft tissues using a special press with a force of 100 kg/1 kg of animal weight. Slices were prepared and glutamate uptake was assayed immediately after compression, and again at 2, 24, and 48 hours of decompression. RESULTS: The level of glutamate uptake in cerebral cortex and hypothalamus slices immediately after compression was reduced by 24% (p<0.05) and 31% (p<0.005) respectively. Glutamate uptake in cerebral cortex and hypothalamus was nearly at the same level as the intact group of animals (without compression). In cerebellum slices, glutamate uptake also increased, and in fact exceeded the level in the intact group by 17% (p<0.025). The level of glutamate uptake decreased in cerebral cortex, hypothalamus, and cerebellum slices by 45% (p<0.001), 28% (p<0.001,) and 25% (p<0.001) respectively, compared to the intact group of animals. CONCLUSIONS: Experimental crush syndrome promotes the reduction of the glutamate uptake level in brain tissue slices, such that cerebral cortex > hypothalamus > cerebellum. PMID- 11859272 TI - Minimal distances between temporal bone structures and their mutual correlations. AB - BACKGROUND: The minimal distances between basic temporal bone structures limit the operating field in ear surgery. The aim of the present study was to perform measurements on the minimal distances between key structures of the temporal bone and to investigate possible correlations between given structures. MATERIAL/METHODS: The author removed 100 temporal bones from subjects of both sexes, across a range of age groups and from both sides of the body. Minimal distances were measured between the following structures: the lateral semicircular canal (SC), the tympanic ring (TR), the hard meninges of the middle cranial fossa (HM), the facial nerve (FN), the upper bulb of the internal jugular vein (JB) and the internal carotid artery (CA), collected in the appropriate pairs. The samples were measured using an operating microscope equipped with a graduated eyepiece. RESULTS: The results are presented in various categories according to age, sex, and side of the body. Five pairs of distances were found to be significantly shorter in younger children (group I) than in older children and adults (group II). CONCLUSIONS: Ear surgery in younger children demands much more care than in older children and adults, especially regarding situations when the jugular bulb is in the operating field. PMID- 11859273 TI - mRNA expression of EGF receptor ligands in atrophic gastritis before and after Helicobacter pylori eradication. AB - BACKGROUND: Epidermal growth factor (EGF) receptor ligands (EGFRL) including transforming growth factor alpha (TGF-alpha), amphiregulin, and heparin binding EGF (HB-EGF) are involved in gastric mucosal repair in chronic gastritis. Their mRNA expression has been shown to be upregulated after Helicobacter pylori (H.p.) eradication but little is known about this gene expression in atrophic gastritis. The purpose of our study was to investigate EGFRL mRNA expression in gastric mucosa of patients with atrophic gastritis before and after H.p.-eradication. MATERIAL/METHODS: Antral mucosal biopsies were obtained during endoscopy in 10 H.p. positive patients with atrophic gastritis and in 10 H.p. negative controls with intact mucosa. Total RNA of antral biopsies was extracted and RT-PCR was performed, the PCR-products being measured densitometrically. Values were compared with mRNA expressions in H.p. negative antral mucosa (n=10). RESULTS: Gastric biopsies revealed mRNA expression for TGF-alpha, amphiregulin and HB-EGF, both in H.p. positive atrophic antritis and in H.p. negative healthy mucosa. The mRNA expression of TGF-alpha in atrophic gastritis was significantly upregulated after H.p.-eradication, whereas that of amphiregulin did not change after this eradication. Expression of HB-EGF mRNA was higher in H.p.-infection than after H.p.-eradication or in H.p. negative healthy subjects. CONCLUSIONS: H.p. positive atrophic gastritis is associated with differential mRNA expression of EGF receptor ligands. H.p.-eradication in this entity leads to unequal changes of these growth factor expressions compared to chronic active gastritis without atrophy. PMID- 11859274 TI - Impact of moderate aerobic exercise training on insulin sensitivity in type 2 diabetic men treated with oral hypoglycemic agents: is insulin sensitivity enhanced only in nonobese subjects? AB - BACKGROUND: Few studies have evaluated the effect of aerobic physical training on insulin sensitivity in subjects with type 2 diabetes treated with oral hypoglycemic agents. Most studies that involved nonobese subjects showed enhanced insulin sensitivity whereas studies involving obese subjects with type 2 diabetes are inconsistent because of concomitant fat loss. MATERIAL/METHODS: Thirteen men with type 2 diabetes (6 nonobese and 7 obese) treated with oral hypoglycemic agents carried out a 12-week ergocycle program at 60% VO2 peak, one hour thrice a week. Diet and medications were maintained throughout the study. Insulin sensitivity was assessed with the euglycemic hyperinsulinemic clamp technique. RESULTS: VO2 peak (ml/kg/min) increased with exercise training. There were no significant changes in fat mass, percent body fat, fasting plasma glucose and insulin levels, glycated hemoglobin levels, or insulin sensitivity. However, when the subjects were separated into two groups, they showed different insulin sensitivity adaptation to training. In fact, nonobese subjects with type 2 diabetes showed an increase in insulin sensitivity, whereas no change was observed in the obese subjects. CONCLUSIONS: These findings suggest that moderate aerobic exercise training of 3 months duration enhanced insulin sensitivity only in nonobese men with type 2 diabetes. Obese men with type 2 diabetes showed no benefit. Thus, adiposity per se may be a determining factor in the exercise induced metabolic benefit of exercise training in subjects with type 2 diabetes. PMID- 11859275 TI - Application of perfluorocarbon liquid in the removal of metallic intraretinal foreign bodies. AB - BACKGROUND: Virectomy is recognized as one of the best effective methods for removing interetinal foreign bodies, but many authors showed complications during or after removing IOFB. The purpose of our study was to show that the use of perfluorocarbon liquid minimizes complications during pars plana vitrectomy for the removal of metallic intraretinal foreign bodies (IRFBs). MATERIAL/METHODS: Fourteen consecutive eyes presenting with penetrating eye injury and intraretinal foreign bodies had previously undergone vitrectomy with injection of perfluorocarbon liquids and removal of IRFBs. In all cases a silicon oil or gas C3F8 tamponade was performed. In one case the removal of the foreign body was combined with repair of retinal detachment using internal tamponade with silicon oil. RESULTS: After a mean follow-up of 16.4 months, 78.5% of the eyes had recovered better than 20/70 vision, and 92.8% had attached retina. One patient had endophthalmitis and phthisis, and perioperative complications were encountered in two cases. In one case there was a limited hemorrhage, intraretinal, while the other complication was a retinal detachment, which was immediately corrected. In the follow-up period endophthalmitis and lens opacity was found in one patient and epiretinal membrane in another case. These postoperative complications were repaired by additional surgery. CONCLUSIONS: The usage of perfluorocarbon liquid is effective for removal of intraretinal foreign bodies, and allows to minimize intraoperative complications. PMID- 11859276 TI - Does schistosomiasis interfere with application of the Knodell score for assessment of chronic hepatitis C? AB - BACKGROUND: Schistosoma mansoni (SM) is a significant cause of liver disease in many countries. Chronic hepatitis C (HCV) is a worldwide health problem. The association of SM and chronic HCV is not uncommon, especially in areas where both diseases occur. The possible synergistic relationship between them is controversial. Also, the degree of necroinflammatory injury and the stage of fibrosis in patients with mixed schistosomiasis and chronic HCV remains unclear. MATERIAL/METHODS: 185 individuals were studied: 25 with pure SM, 100 with pure HCV, and 60 mixed HCV and SM (HCV+S), selected from 222 biopsied patients with chronic liver disease treated at the liver unit of the Internal Medicine Department, Mansoura University (July 1999-May 2000). They were subjected to rectal snip and serological test for schistosomiasis, liver functions, HBV, HCV serological markers, serum qualitative PCR, and liver biopsy. Masson trichrome stain was performed to assess fibrosis. Immunohistochemical staining was performed for HBsAg and HBcAg. The Modified Knodell score was applied to assess the biopsy. RESULTS: Five of the 25 pure SM and 2 of the HCV+S group revealed schistosomal granuloma. 30% of the pure HCV and 33% of the HCV+S patients were found to be cirrhotic. No statistically significant difference was identified between the groups as regards necroinflammatory injury or Knodell score, or the stage of fibrosis. CONCLUSIONS: Schistosomal hepatic infection does not lead to more severe or progressive disease in patients with chronic hepatitis C. PMID- 11859277 TI - Early cardiovascular involvement in patients with systemic sclerosis (SSc). AB - BACKGROUND: Systemic sclerosis (SSc) is a connective tissue disease characterized by progressive fibrosis of the skin, and by distinctive forms of internal organ involvement. Symptoms usually occur relatively late and are nonspecific, thus often unrecognized. The purpose of our study was to evaluate cardiac function in patients with SSc by means of non-invasive methods in order to detect early dysfunction of the cardiovascular system. MATERIAL/METHODS: A group of 22 patients with SSc was compared with a group of 22 healthy volunteers. Standard EKG, 24-hour Holter monitoring (HM), and echocardiography were performed in all subjects. RESULTS: In patients with SSc, HM revealed a tendency to tachycardia. Conduction disturbances were observed in 3 patients. In 6 patients significant ventricular arrhythmia was found. Silent ischemia episodes were detected in 6 patients. In HRV analysis, significantly lower values were detected in patients with SSc than in controls. LPs were present in one patient with SSc, and none in the control group. The mean values for QT interval did not exceed the range of normal values. No signs of systolic cardiac dysfunction were detected, while in 6 patients left ventricle diastolic dysfunction was recognized. Valvular lesions were observed in 8 patients, but only in 2 patients were they hemodynamically important. CONCLUSIONS: 24-hour Holter monitoring and echocardiography enable the early detection of cardiovascular dysfunction in patients with systemic sclerosis presenting without apparent cardiac impairment symptoms. PMID- 11859278 TI - Clinical signs pointing to the source of hemorrhage in multiple intracranial aneurysms. AB - BACKGROUND: In multiple aneurysms of cerebral vessels it is essential to correctly identify the aneurysm that has caused the hemorrhage, since it must be operated first. On the basis of our own material, we have evaluated the usefulness of various clinical signs and examinations in diagnosing which of several aneurysms was the cause of an intracranial hemorrhage. MATERIAL/METHODS: The material consisted of 163 patients with a total of 391 aneurysms. Diagnosis was based on panangiography and neurological examinations in all cases and CT in the majority. RESULTS: All the diagnostic examinations in question (neurological, angiography, CT and intraoperative evaluation) enabled a correct diagnosis in 72.6% of cases. In CT scanning, diagnostic value diminishes with time. Intracerebral hematoma is the most persistent sign. The highest rupture index involved aneurysms localized on the anterior communicating artery, while those on the internal carotid artery ranked second. Four patients in whom intraoperative evaluation showed that the order in which the aneurysms were treated surgically was erroneous died as the result of rebleeding that occurred prior to planned follow-up surgery. CONCLUSIONS: It is vitally important for outcome to determine which of several aneurysms has ruptured, since errors in the sequencing of surgical repair significantly increase mortality due to rebleeding preceding the next stage of surgery. When diagnosing the source of a hemorrhage one should take into account neurological findings, angiography, EEG, CT, MRI, and the location of the aneurysms. PMID- 11859279 TI - Is there a relationship between left ventricular systolic function and serum cytokines level in patients with coronary artery disease? AB - BACKGROUND: There is growing evidence that cytokines play a pathogenic role in both heart failure and atherosclerosis. The aim of our study was to assess serum levels of selected cytokines and soluble forms of receptors in patients with coronary artery disease (CAD), and the relation between these concentrations and left ventricular (LV) systolic function. MATERIAL/METHODS: We measured serum levels of TNFalpha, sTNFR 1 and 2, IL-2, and IL-10 in 100 patients with CAD: 70 patients with LV regional wall motion disturbances (group A); and 30 patients with normal LV function (group B). The control group (group C) consisted of 20 healthy volunteers. RESULTS: Mean serum TNFalpha and IL-10 concentrations were significantly higher in groups A (18.3 +/- 3.5; 55.4 +/- 118.5 pg/ml) and B (17.9 +/- 4.9; 45.3 +/- 76.8 pg/ml) than in controls (8.3 +/- 1.4, p<0.001; 14.3 +/- 28.5, p<0.05; respectively). Moreover, in group A serum levels of sTNFR 1 were higher (1367.4 +/- 531.1 pg/ml) than in group C (1093.9 +/- 456.9 pg/ml; p<0.05). No differences were found in the study groups between serum sTNFR 2 and IL-2 levels. In group A, both the LV ejection fraction and motion score index correlated with TNFalpha (r=-0.277; r=0.282; p<0.05), and sTNFR 1 levels (r= 0.258; r=0.280; p < 0.05). CONCLUSIONS: The serum concentrations of TNFalpha and IL-10 are increased in patients with CAD. Additionally, patients with impaired LV contractility have higher sTNFR 1 concentrations. Serum cytokine activation may play a role in the development of heart failure in patients with CAD. PMID- 11859280 TI - Modified neck dissections--efficiency of surgical treatment and clinical observations. AB - BACKGROUND: The evaluation of affected lymph node number, the site of metastatic lymph nodes, and spread of neoplastic infiltration beyond the lymph capsule is considered useful in prognosis. The aim of the study was to estimate the frequency of neck nodal metastases occurrence depending on the site of origin, the grade of larynx cancer progression, and clinical condition of neck lymph nodes. MATERIAL/METHODS: The study comprised 315 patients with larynx cancer who underwent surgery in 1994-1999. Laryngectomy was the most frequent procedure, performed in 254 patients, while partial laryngectomy was performed in 61 patients. There were 630 bilateral neck dissections; type I of modified radical surgery in 27 cases, type II in 45 cases, and type III in 558 cases. RESULTS: Neck nodal metastases were detected most rarely in cases of glottic cancer (16%), and most frequently in transglottic cancer (56%). Most frequently, the nodal metastases affected the II level of the lymph node groups regardless of the site. Recurrences to the neck lymph nodes were observed in 3.3% of the cases where the removed lymph nodes did not reveal any metastatic changes in histopathological examinations. CONCLUSIONS: Postoperative results of the neck lymph node histopathological examination point to the necessity of lymphadenectomy in the II V levels. Selective lymphadenectomy is not advisable due to the possibility of metastases to all levels of the neck lymph nodes regardless of the primary tumor site. PMID- 11859281 TI - Vitamin C concentration in gastric juice in patients with precancerous lesions of the stomach and gastric cancer. AB - BACKGROUND: The aim of the study was to elucidate what are the concentrations of vitamin C in gastric juice in patients with gastric cancer in comparison to patients with metaplasia. MATERIAL/METHODS: In patients aged 20 to 75 years with H. pylori infection and chronic gastritis, metaplasia and gastric cancer the concentration of vitamin C was determined by spectrofotometry of gastric juice during gastroscopy. On the basis of the results of histological examination the following four groups were isolated. Group I (control)--12 patients with normal gastric mucosa, group II--15 patients with chronic gastritis, group III--17 patients with metaplasia and group IV--16 patients with gastric cancer H. pylori infection was confirmed by urease test and histological examination (Giemsa stainning) in all patients. RESULTS: In controls the mean concentration of vitamin C in gastric juice was 18.2 mg/ml (5.7-31.2 mg/ml), in group II--6.3 mg/ml (2.9-13.1 mg/ml) in group III--3.9 mg/ml (2.6-10.1 mg/ml) and group IV--3.2 mg/ml (1.7-9.2 mg/ml). Statistically significant differences of vitamin C concentration were found among group I and group II, III and IV (p<0.001) and among groups II and III and IV (p < 0.01). CONCLUSIONS: There are not differences of vitamin C concentration in gastric juice between patients with metaplasia and patients with gastric cancer. This points out that low levels of this vitamin in gastric juice might play the role in the earlier stages of carcinogenesis. PMID- 11859282 TI - Cytomegalovirus infection and atherogenesis. AB - BACKGROUND: Experimental studies and clinical observations indicate the importance of active CMV infection in the development of atherosclerosis. Our research constituted an attempt to evaluate the components of lipid metabolism (total cholesterol and its fractions, triglycerides) in children with active CMV infection. MATERIAL/METHODS: The research involved 45 children with active primary or secondary CMV infection. The diagnosis was established by measuring anti-CMV serum antibodies (IgM and IgG) using the Enzyme-Linked Fluorescent Assay (ELFA) technique, and by detecting CMV-DNA using the PCR technique. On the basis of diagnostic tests, the children were allocated to one of two groups. The first group of 15 children manifested primary active infection with CMV. The other group consisted of 30 children diagnosed with secondary active CMV infection. RESULTS: In the first group, the mean values of the serum lipid profile were as follows: total cholesterol--142 +/- 32.9 mg/dl; HDL-cholesterol:-- 23.4 +/- 8.6 mg/dl; LDL-cholesterol--97.8 +/- 30.9 mg/dl; triglycerides--83 +/- 15.5 mg/dl. In the second group of children, the mean values of serum lipid profile were as follows: total cholesterol-- 142.5 +/- 44.2 mg/dl; HDL-cholesterol--21.4 +/- 10.8 mg/dl, LDL-cholesterol--87.1 +/- 35.3 mg/dl; triglycerides--101.7 +/- 31.2 mg/dl. In all these children, the values of total cholesterol, LDL-cholesterol, and triglycerides conformed to published norms, while HDL-cholesterol was significantly below normal. CONCLUSIONS: The results we obtained point to dyslipidemia in the course of active CMV infection, which may be of pathogenetic significance at any stage of atherogenesis. PMID- 11859283 TI - Serum levels of tissue inhibitor of metalloproteinases 2 in systemic sclerosis: a preliminary study. AB - BACKGROUND: The etiopathogenesis of systemic sclerosis (SSc)--an autoimmunological disease characterized by excess collagen and other connective tissue components in skin and internal organs--remains unclear. Given the potential role of matrix enzymes, metalloproteinases, and their tissue inhibitors in pulmonary fibrosis, we assayed the serum concentration of tissue inhibitor of metalloproteinases 2 (TIMP-2) in patients with SSc and explored its possible correlation with the duration of Raynaud's phenomenon, the intensity of sclerosis of the skin, and pulmonary lesions. MATERIAL/METHODS: We studied 18 SSc patients, 8 with limited SSc and ten with diffuse. The degree of sclerosis of the skin was measured using the Total Skin Scale. Chest x-rays and spirometric examinations were also performed. The presence and type of antinuclear antibodies (ANA) were determined by indirect immunofluorescence and double immunodiffusion in agar gelatin. Serum TIMP-2 concentration was measured by ELISA. RESULTS: The average serum TIMP-2 concentration was not significantly higher in SSc patients than in controls. No statistically significant difference was found in this respect between limited and diffuse SSc. Despite the lack of correlation between the duration of Raynaud's phenomenon, the degree of sclerosis of the skin, and the serum concentration of TIMP-2, the latter concentration was higher in patients with restrictive pulmonary dysfunction in spirometric testing. CONCLUSIONS: Our research, though involving a small group of patients, points to the probable role of TIMP-2 in the development of pulmonary fibrosis. PMID- 11859284 TI - Total IgE levels in children under three years of age. AB - BACKGROUND: The course of type 1 allergic reaction is a result of complex immune processes in which immunoglobulin E plays a crucial role. The serum level of this immunoglobulin (IgE(total)) is closely related to the age of the patient. The presented paper analyses IgE(total) in children below 3. MATERIAL/METHODS: The study was carried out in group of 291 patients aged 5-39 months. IgE(total) were determined using the Fluoro-FAST 3M. The analysis took into consideration the age, sex and living place of the examined children. RESULTS: The artmetic mean of IgE(total) was 38.8 +/- 4.0 IU/ml, the median--12 IU/ml, whereas the 5th and 95th percentile values amounted to 1IU/ml and 185 IU/ml, respectively. According to the standards recommended by the manufacturer of reagents used for determination of IgE(total) (Bio Whittaker), 103 children (35.4%) had elevated serum levels of IgE(total), whereas 196 (67.4%) were found to have elevated IgE(total) according to population standards. The obtained percentage distribution of elevated IgE(total) values in particular age groups is characterized by stepwise changes, observed in the age intervals for which the range of values considered as normal by the manufacturer is changed. CONCLUSIONS: On the basis of analyzed IgE(total) determinations it has been established that the manufacturer's standards for children aged from 2 to 3 years are overestimated, whereas the population standards seem to be underestimated. Therefore, the determination of IgE(total) normal value range requires further studies. PMID- 11859285 TI - Evaluation of lymphocyte subsets and NK cells in septic children. AB - BACKGROUND: Disorders of lymphocyte subpopulations counts and NK cells may not only contribute to impaired elimination of pathogens, but also be an important factor in the development of generalized inflammatory reaction. The aim of the paper was the analysis of subpopulations of lymphocytes and NK cells in children with sepsis and after recovery. MATERIAL/METHODS: The study comprised 25 septic children (17 infants and 8 children aged from 1 to 3 years). The control group consisted of 25 healthy children. The study was conducted in the initial phase of sepsis and then repeated 2 to 3 months after recovery. In all the children, the percentage and absolute numbers of basic lymphocyte subsets and NK cells in blood serum were evaluated. RESULTS: In the course of sepsis, the absolute number of activated lymphocytes T in the youngest children was higher in comparison with the control group. After recovery, the percentage and absolute counts of CD4 lymphocytes in all the children and CD3/CD4 lymphocytes ratio in the youngest children were considerably lower than in the control group. The percentage and absolute number of NK cells in the youngest children after recovery were higher in comparison with the control group. CONCLUSIONS: In children who underwent sepsis, depression traits of cellular immunity are observed. The occurrence of defensive system disorders after recovery may be an important factor in recurrent infections and creates the necessity of immune system monitoring in children with the history of sepsis for the purpose of prevention. PMID- 11859286 TI - Distribution of hepatitis C virus RNA in whole blood of patients with HCV infection and HBV-HCV coinfection. AB - BACKGROUND: Although HCV and HBV are essentially hepatoptropic, several lines of evidence suggest that these viruses can infect other cells, also PBMC in most patients with chronic HCV. MATERIAL/METHODS: The presence of HBV DNA and HCV-RNA was determined by a polymerase chain reaction (multiplex PCR and RT-PCR - nested PCR) in a group of patients with chronic liver disease. HCV-RNA was investigated in serum, plasma and peripheral blood mononuclear cells (PBMC) while HBV-DNA only in serum or plasma. RESULTS: Among 374 patients tested, HCV-RNA was detected in the venous blood of 208 patients; HCV RNA alone was detected in 154 patients and 54 patients were co-infected by HCV and HBV. HBV-DNA was found in 128 of 374 patients, while infection by HBV only was found in 74 patients. It was also shown that in the presence of HBV the replication ability of HCV is lower (p=0.085, Goodman-Kruskal Gamma = 0.561 and YuleQ = 0.5610). CONCLUSIONS: Since coexistence of HBV and HCV is not a rare case, diagnostics of hepatitis cannot be limited to detection of one type of the virus only. Misinterpretation of the virus type that caused the infection may lead to serious complications, especially in those cases when interferone is used for treatment. PMID- 11859287 TI - The course of vision disturbances in a patient with the MELAS syndrome. AB - BACKGROUND: This article describes the neuropsychological and ophthalmic symptoms presented by a patient with MELAS, a mitochondrial cytopathy. This rare disease is characterized by a remitting-relapsing course against the background of a slowly progressive degenerative process. CASE REPORT: The patient is a 22-year old Polish female, with initial onset of symptoms in 1994; the clinical diagnosis of MELAS was established in 1998, and confirmed in 2000 by the discovery of a novel mtDNA mutation. Her visual acuity fluctuates from near-normal to near blindness, often changing dramatically within a matter of weeks; the visual field has more or less steadily narrowed to lunate. Visual evoked potentials show sporadic disturbances, while the nerve fiber layer shows significant attenuation. The evidence points to a complex etiology, involving both cortical damage and attenuation of the optic nerves and neural pathways. A similar two-phase pattern- episodic disturbances with rapid spontaneous recovery against a background of progressive deterioration--occurs in neuropsychological testing, which reveals progressive dementia and episodic aphasia. CONCLUSIONS: The peculiar pathomechanism of MELAS results in simultaneous insults to various parts of the central and peripheral nervous systems, creating the complex and highly variable pattern seen in this patient. In clinical practice care should be taken not to overlook the possible significance of such a pattern appearing in various systems and on varying levels. PMID- 11859288 TI - Tumor markers in metastatic disease from cancer of unknown primary origin. AB - BACKGROUND: About 5-10% of cancer patients suffer from cancer of unknown primary (CUP). Sophisticated diagnostic techniques have not improved diagnostic efficacy for these patients. The aim of our analysis was to evaluate serum levels of four tumor markers and to relate them to the histology, number, and sites of metastases, the response to chemotherapy, and survival. MATERIAL/METHODS: Blood samples from 46 patients with CUP were assayed for CEA, CA 19-9, CA 15-3 and CA 125. Specific IMx tumor marker assays (Abbott) were used to measure tumor marker quantities. RESULTS: No significant differences were detected between the mean levels of all markers for the two histological types (adenocarcinoma vs. undifferentiated carcinoma). The comparison of tumor marker values in patients with one metastatic location to those with two or more showed no significant differences. We found no differences in marker values between patients with disease sites in the thorax vs. those with abdominal metastases. The comparison of marker negative, CEA positive and non-CEA positive patients failed to reveal significant differences in response rate. The comparison of CEA positive and marker negative patients showed significant differences in survival (log-rank, p=0.02). CONCLUSIONS: Our results suggest that patients with CUP have a nonspecific overexpression of serum tumor markers and that routine use of these markers does not offer diagnostic assistance. Nevertheless, the pattern of expression of some biological markers, such as CEA, could have prognostic significance for survival. PMID- 11859289 TI - Dynamic MR imaging of soft tissue tumors with assessment of the rate and character of lesion enhancement. AB - BACKGROUND: The aim of this study was to analyze the diagnostic usefulness of dynamic MRI with determination of the coefficient of enhancement rate and the character of tumor enhancement, and to assess both parameters in the differentiation of malignant lesions. MATERIAL/METHODS: The material consisted of 45 patients (30 sarcomas, 15 non-malignant lesions), age 16-64 years. MRI was done using an Elscint 2T unit, gradient echo techniques, apex angle 80 degrees. The repetition time (TR) was 80-200 ms, the echo time (TE) was 2-6 ms, 1 excitation; the acquisition time (TA) was 70-80 ms. The coefficient of tissue enhancement rate was calculated in the region of interest, and expressed as percent per second (erc%/s). The limit value of erc%/s was determined. The sensitivity and specificity of MRI were calculated in the differentiation of malignant tumors. The method of contrast filling of the tumors was assessed in successive phases after administration of gadolinium Gd-DTPA. RESULTS: Dynamic MRI with determination of the index of tumor enhancement rate is highly sensitive (93%) and specific (73%) in the differentiation of malignant and benign lesions. The usefulness of the assessment of tumor enhancement character was not confirmed, since the sensitivity and specificity were 73% and 33%. Dynamic MRI with determination of erc%/s and tumor enhancement character is highly sensitive (93%) and specific (87%). CONCLUSIONS: Dynamic MRI with determination of erc%/s and tumor enhancement character is the best method for differential diagnosis. PMID- 11859290 TI - MR proton spectroscopy in liver examinations of healthy individuals in vivo. AB - BACKGROUND: Magnetic resonance proton spectroscopy ((1)H MRS) is a non-invasive method that provides in vivo measurement of metabolic concentrations in body tissue. However, relatively little is known about the potential of 1H MR spectroscopy for the quantification of liver metabolites. The aim of this study was to determine the usefulness of (1)H MRS in establishing the metabolic pattern of normal human liver. MATERIAL/METHODS: Proton spectroscopy of the liver, using the Picker system, Edge Eclipse 1.5T, with whole-body coil and PRESS 35 sequence, was conducted in a group of 24 healthy volunteers. In all subjects we also evaluated the thickness of the subcutaneous fat tissue in MR images and body mass index. RESULTS: The presence of lipid resonances, phosphoesters (Pe), glycogen/glucose (Glc), and glutamine/glutamate (Gln) were observed in the spectra. Total lipid concentration, CH3 and (CH2)n of the lipid resonances were higher in the men than in the women, while the metabolite contents in the CH2=CH CH2 groups of lipids, Pe, Glc and Gln peaks were similar in both genders. We observed statistically significant positive correlation, more apparent in the group of men, between TL concentration and BMI. There was no statistically significant correlation between lipid total concentration and age. CONCLUSIONS: Proton spectroscopy enables the quantitative evaluation of lipid contents in the liver. The correlation between liver triglyceride contents and body mass index shows that the tendency to obesity is also connected with lipid accumulation in the liver. PMID- 11859292 TI - Clinical outcomes of silicon carbide coated stents in patients with coronary artery disease. AB - BACKGROUND: According to the literature, the stent thrombosis and restenosis rate after coronary angioplasty (PTCA) can be reduced by using stents covered with biologically active substances. The aim of our study was to investigate clinical outcomes of PTCA alone, conventional stainless steel stents, and special Tenax stents covered by amorphous silicon carbide (a-SiC: H). MATERIAL/METHODS: The study involved a retrospective cohort of 300 patients with coronary artery disease who underwent interventional treatment from 1998 to 2000. One hundred patients received a Tenax stent (BIOTRONIK, Germany) covered by a-SiC: H. For every patient who received a Tenax stent, one patient with non-Tenax stenting and one patient with PTCA without stenting were randomly assigned. RESULTS: In the Tenax group, 1% of the patients had an early coronary event due to subacute stent thrombosis, as compared to 2% in the non-Tenax group and 3% in the PTCA group. At 6-month follow-up, the difference in the frequency of major adverse cardiac events was more express between the PTCA group and both groups of patients with stents (32% vs. 20%; p=0.053; 32% vs. 14%; p=0.002) in comparison with the 30-day and 3-month results. In the non-Tenax stenting group more events were detected than in the Tenax group (20% vs. 14%; p=0.259), but the difference was not statistically significant. CONCLUSIONS: The results strongly suggest the protective qualities of a-SiC: H stents in reducing early and late coronary events. It can be speculated that Tenax stents attenuate the progression of endothelial growth at the site of intervention. PMID- 11859291 TI - An open-label study to evaluate the safety, tolerability and efficacy of rivastigmine in patients with mild to moderate probable Alzheimer's disease in the community setting. AB - BACKGROUND: Long-term safety and efficacy of Exelon (rivastigmine) was evaluated in a multi-center open-label study of 62 patients with probable mild to moderate Alzheimer's disease living in community setting. MATERIAL/METHODS: The patients started treatment with 1.5 mg bid (3 mg/day) Exelon and were scheduled to receive doses of 1.5 mg bid Exelon escalating on a biweekly basis. The patients were maintained on the highest tolerated dose within the assigned dose range 1.5-6.0 mg bid (3-12 mg/day) for the rest of the study. Evaluations were scheduled at biweekly intervals for the first 8 weeks and subsequently at study weeks 12, 18 and 26. Effects of Exelon on cognition were evaluated using the mini-mental state examination (MMSE) and selected items of Alzheimer's disease assessment scale (ADAS-cog) and the staging of the disease was measured using the global deterioration scale (GDS). Safety was monitored by physical examinations, vital signs, laboratory tests, ECG recording and by the assessment of adverse events. RESULTS: 55 patients completed the study (89%). Patients treated for 26 weeks showed the mean MMSE, ADAS-cog and GDS scores close to baseline values (p=NS) with no improvement and no deterioration. Exelon was generally well tolerated with 11% of patients withdrawing due to adverse events. The most frequently reported adverse events related to the gastrointestinal tract. CONCLUSIONS: In conclusion, the study data indicate that treatment with Exelon is safe, generally well tolerated and inhibits the progression of cognitive decline in patients with mild to moderate Alzheimer's disease over 26 weeks of treatment. PMID- 11859293 TI - Tonal nitric oxide and health: anti-bacterial and -viral actions and implications for HIV. AB - Nitric oxide has been shown to have important physiological regulatory roles, i. e, vasodilation, neurotransmitter release, etc. Now, we review its role as an antibacterial and antiviral agent. Nitric oxide has also been identified as an important factor in the development of non-specific immunity. And accordingly, nitric oxide synthase (NOS), the catalytic enzyme producing nitric oxide, is a key element in the protective activities of nitric oxide. The expression of inducible (i) NOS is regulated by cytokines. iNOS-derived nitric oxide was found to contribute to both early and late phases of antibacterial activity. Enzymes, such as proteases (reverse transciptases, and ribonucleotide reductase, etc.) containing cysteine residues, appear to be targets for nitric oxide nitrosylation, as well as viral-encoded transcription factors that are involved in viral replication. It would appear that this multifunctional signaling molecule is not only involved with signaling between cells, it also appears to maintain the immediate environment free of microbial agents. PMID- 11859294 TI - Hepatocyte proliferation in health and in liver failure. AB - Mammalian liver possesses an extraordinary capacity for compensatory growth in response to conditions that induce cell loss by physical, infectious, or toxic injury. In normal animals and humans, it is a tightly regulated process of both hypertrophy and hyperplasia involving different liver cell populations and a finely tuned interplay between growth factors, cytokines, extracellular matrix components and other regulators. The regeneration response is maximal when two thirds of the liver is resected. When a lesser amount of parenchyma is removed, residual liver grows more slowly. Resections exceeding two-thirds of the liver mass also retard and diminish both DNA synthesis and mitotic activity and subtotal (90%) hepatectomy invariably results in the death of rats without regeneration. The underlying mechanisms of liver growth inhibition are poorly understood. In particular, only a few studies exist that provide insight into the mechanisms that control regeneration after extensive hepatocyte loss. In this regard, the role of growth-regulatory factors and other compounds that accumulate in the blood circulation as a result of hepatic insufficiency and liver cell death remains unclear. We have initiated studies of these mechanisms and demonstrated that in rats with low (10%) hepatocyte mass, a marked and sustained elevation of blood IL-6, HGF and TGF-b1 levels was associated with lack of hepatocyte proliferation and suppression of Stat3 DNA binding. While searching for the possible cause of inhibited IL-6/Stat3 signaling, we found that IL-6 receptor (IL-6R & gp130) was preserved, that nuclear Stat3 protein content was lowered, and that IL-6/Stat3 pathway inhibitors (SOCS-1, PIAS3) were induced during the pre-replicative Go-G1 period. PMID- 11859295 TI - Regulation of renal tubular sodium transport by cardiac natriuretic peptides: two decades of research. AB - This review presents the current state of our knowledge regarding the regulation of renal tubular sodium transport by natriuretic peptides, with special emphasis on recent findings in this field. Natriuretic peptides constitute a complex system involved in the regulation of sodium balance and blood pressure. The natriuretic peptide family consists of atrial peptides, such as atrial natriuretic factor (ANF, ANP(99-126)), long-acting natriuretic peptide (ANP(1 30)), vessel dilator (ANP(31-67)) and kaliuretic peptide (ANP(79-98)), as well as brain or B-type natriuretic peptide (BNP), C-type natriuretic peptide (CNP) and urodilatin. Natriuretic peptides act on target cells through A-type and B-type receptors and stimulate cyclic GMP synthesis. ANF stimulates natriuresis mainly by inhibiting sodium reabsorption in the inner medullary collecting duct. The effect results from coordinate inhibition of apical sodium channels and basolateral Na+, K+-ATPase. Additional effects on sodium transport occur in more proximal nephron segments and on glomerular filtration when hormone concentration is elevated. BNP and urodilatin share the same mechanism of action. CNP synthetized in several nephron segments acts through specific B-type natriuretic peptide receptors, which are also expressed in renal tubule, but have a different distribution than A-type receptors. ANP(1-30), ANP(31-67) and ANP(79-98) decrease Na+, K+-ATPase activity in tubular cells through a prostaglandin E2-dependent mechanism. PMID- 11859296 TI - Managing resources in radiology: a necessary philosophy of work. PMID- 11859297 TI - Guidelines in diagnostics: formulation methodology proposal. AB - In healthcare the term "guideline" is used in numerous circumstances and for different purposes. The most commonly accepted definition is provided by the American Agency for Healthcare Research and Quality, which defines guidelines as "recommendations developed in order to assist doctors and patients in the decision-making process, while choosing an appropriate assistance in specific clinical circumstances". At present, in Italy and foreign countries, guidelines are considered an effective tool. Guidelines contribute with a qualitative improvement in performances, standardizing healthcare criteria, enabling also rationalisation and optimisation in both human and economical resources. The achievement of these aims depends on a thorough examination of both meaning and role played by guidelines in clinical practice, and of the essential methodological criteria of their processing. Unfortunately, these objectives can be attained only after the overcoming of obstacles, due to both limits in the formulation of such documents (use of inadequate methodology, choice of the paper based collection of documents rather than computer-based data-processing support), and to the opposition of the medical personnel. The latter problem being caused by a poor knowledge of the role played by guidelines and by the difficulty of a critical evaluation of their validity. In fact, the success of a guideline is also determined by its development, implementation and introduction strategies, and by the consence of the medical community. Once formulated by an interdisciplinary group of experts, a guideline should be necessarily revised according to already coded procedures. A reliable contribution to the diffusion of such guidelines derives from the use of computer-based data-processing supports, which contribute to their interactivity, user friendliness, user adaptivity and updatebility. The above illustrated procedures represent an unquestionable contribution to the clinical decision-making process. In fact, these represent the basic development tools for the harmonisation of cultural qualification and clinical practice, aiming at quality improvement and rationalisation in health-related activities. PMID- 11859298 TI - Acute spontaneous aortic pathologies associated with intramural hematoma: can a differential diagnosis be made by imaging techniques? AB - Acute intramural aortic hematoma may result from ruptured vasa vasorum without intimal disruption (IMH hereafter) but can also be found in association with processes in which an intimal lesion is present, such as aortic dissections (AD), where the hematoma represents the acute thrombosis of the false lumen, or in penetrating atherosclerotic ulcers (PAU), where blood collects beyond the disrupted internal elastic lamina. As the clinical presentation of these conditions is very similar, imaging techniques should aim both to diagnose the hematoma and to characterize the underlying pathology. This review critically analyzes the diagnostic potential of several imaging methods based on personal experience and literature reports. Although intramural hematoma can be easily diagnosed unrespective of the tomographic imaging modality, it is usually difficult to distinguish between IMH and hematomas associated either to AD or to PAU. Indeed, only PAU related hematomas can be consistently diagnosed due to the high probability of unveiling the causative wall ulceration; conversely, the differential diagnosis between IMH and AD - associated hematomas is still matter of controversy. According to the literature, the two conditions cannot be differentiated by transesophageal echocardiography, though some reports state that a distinction might be made by MR imaging. As regards CT, only a small percentage of AD - associated hematomas exhibit clear evidence of a typical intimal flap, allowing for a safe diagnosis to be made: often, unfortunately, the intimal lesion subsequently found at surgery cannot be detected, yielding a picture of IMH. Therefore, the radiological report should give the same diagnostic probability to both conditions. PMID- 11859299 TI - Cardiac MRI: comparison between single-shot fast spin echo and conventional spin echo sequences in the morphological evaluation of the ventricles. AB - PURPOSE: Black blood single shot FSE sequences (Nffse) employ 180 degrees RF refocalisation pulses preceded by an inversion RF double pulse associated to presaturation pulses. The latter produce signal void of the external volume, and possible reduction of the field of view without wrap-around artifacts along the phase coding direction. The aim of our study was to compare the diagnostic possibilities of the Nffse sequences with those of conventional SE study of cardiac morphology. MATERIAL AND METHODS: Twenty-five patients (19 males and 9 females with age ranging from 20 to 54 years) presented findings suggesting right ventricular arrhythmogenic dysplasia. MR examinations were performed with a 1,5 T unit (GE Signa Horizon Echospeed 8.3, Milwaukee, USA) and Torso Phased Array coil positioned at thoracic level. The morphologic study was performed with SE multiphase-multislice ECG-gated sequences (TR: R-R, TE: 30 ms, FOV 320X250, matrix 160X256, slice thickness 10 mm, acquisition time about 5 minutes) and Single-Shot FSE Half Fourier sequences (TR: R-R, TE: 30 ms, flip angle 120 degrees, ETL 30-40, FOV 360X180, Phase FOV 0,5, VBW 64 MHz, slice tickness 10 mm, acquisition time about 10-12 seconds), by imaging along the long and short axis. The study was completed with Fast Gradient Echo sequences (TR: 9ms, TE: 8,2ms, flip angle 25 degrees, VBW 15,63 MHz, FOV 320X250, 10 mm slice thickness, matrix 128X256), subsequently assessed by cine-MR. In order to compare both sequences, two experienced radiologists performed an analysis of quantitative parameters (signal intensity ratio between fat and muscular interventricular septum) and qualitative parameters (double blind evaluation for the presence of cardiac and respiratory artifacts). RESULTS: The signal intensity ratio for the Nffse sequence images was 4.63 +/- 1.56 on the long axis and 7.69 +/- 2.46 on the short axis, whereas it was 3.17 +/- 0.64 on the long axis and 3,50 +/- 0,75 on the axis one for SE images, with a statistically significant difference (p<0,001 and p<0.002 for the long and short axis, respectively). The two radiologists evaluation of the magnitude of artifacts on the SE and Nffse images was similar only as regards the images with significant artefacts alone. Nffse images consistently afforded a detailed evaluation of the right ventricular wall, although blurring artifacts were more common than with good quality SE images. Presence of fatty infiltration of the right ventricle wall was observed in 5 out of 25 patients. In the remaining 20 patients no fatty substitution of the muscular wall of the right ventricle was observed. DISCUSSION AND CONCLUSIONS: The Nffse sequences provide a number of gated multiphase-multislice images, similar to that obtained by conventional SE sequences, in one breath-hold time interval. Due to high intrinsic contrast and reduction of motion artifacts, the Nffse sequences allow a good evaluation of the ventricular morphology and subepicardial and paracardiac adipose tissue. Image quality can be suboptimal due to blurring artifacts. Therefore Nffse sequences can be advantageously employed to image patients with suspected right ventricular arrhythmogenic dysplasia, whenever conventional SE images exhibit substandard quality. PMID- 11859300 TI - Perfusion MRI of the lung: preliminary results in twenty healthy volunteers. AB - PURPOSE: This study attempted to assess the feasibility and the accuracy of lung contrast enhanced magnetic resonance (MR) perfusion imaging in the evaluation of 20 healthy volunteers and to correlate the intensity/time (I/T) curves with the pulmonary physiological criteria. MATERIAL AND METHODS: After an informed consensus, twenty no-smoker subjects, without any present or past pulmonary pathologies, were studied by means of an ultra-fast gradient echo sequence after a Gd-DTPA intravenous administration. The acquisitions were performed in coronal and sagittal planes, in supine and prone position, in breath holding. An ulterior acquisition was performed in only five cases immediately after 2-3 minutes of physical exercise. RESULTS: All the examinations, reviewed according to a visual score by three independent observers, resulted fairly interpretable. The I/T curves, obtained in post-processing evaluation, have the same trend in all cases and are correlated to the lung physiological criteria: the position changes of the examined subjects confirm the gravity dependence of perfusion; the study performed immediately after physical exercise showed a reduction of perfusion in the whole lung due to a transitory massive shift of blood mass from visceral to muscular districts. CONCLUSIONS: The extremely short acquisition and post processing time coupled with a simple feasibility, the ability to detect physiological perfusional changes, the minimal invasivity indicate a potential clinical use of MR in the pulmonary perfusion defects evaluation. PMID- 11859301 TI - Pancreas divisum and santorinicele: assessment by dynamic magnetic resonance cholangiopancreatography during secretin stimulation. AB - PURPOSE: To evaluate the usefulness of MRCP, before and after secretin administration, in diagnosing Santoriniceles in patients with pancreas divisum. MATERIAL AND METHODS: One hundred and eight patients with suspected pancreatic disease, underwent dynamic magnetic resonance cholangiopancreatography (MRCP) before and after secretin administration (S-MRCP). S-MRCP images were evaluated for presence/absence of pancreas divisum, Santorinicele; size of the main pancreatic duct and of the Santorinicele. The onset of duodenal filling was calculated on dynamic S-MRCP images. S-MRCP findings were compared to endoscopic retrograde cholangiopancreatography (ERCP) ones (39/108). RESULTS: Pancreas divisum (PD) was detected in 6/108 patients (6%) at MRCP, and in 16/108 patients (14%) at S-MRCP. ERCP confirmed the diagnosis in 12/16 patients, with 1 false positive. 3 patients did not undergo ERCP. Santorinicele was detected in 4/108 (3%) patients at MRCP and in additional 4/108 (3%) patients at S-RMCP, only in patients with PD. Santoriniceles were confirmed in 7/8 patients at ERCP; in 1/8 patient CPRE was unsuccessful. The duct of Santorini was significantly larger (p< 0.05), in the pancreatic head, in patients with PD and Santorinicele (3.6 mm) compared to those with PD only (2.2 mm). A significant reduction in size of the pancreatic duct (26%) and of the Santorinicele (63%) was observed after sphincterotomy. The onset of duodenal filling was significantly delayed in patients with Santorinicele (2.1 vs 1.3 minutes)(p<0.05). CONCLUSIONS: S-MRCP helps to identify patients with pancreas divisum and Santorinicele, a known cause of impeded pancreatic outflow which benefits from endoscopic treatment. PMID- 11859302 TI - Endoscopic ultrasound and Computed Tomography in gastric stromal tumours. AB - PURPOSE: Gastric stromal tumors (GIST) have no well defined biological characteristics, from either the pathological or immunohistochemical point of view, which can make their definition by imaging techniques difficult. We evaluated CT and EUS morphologic characteristics and signs of malignancy of eleven cases of GIST and compared the findings to the pathological classification. MATERIAL AND METHODS: EUS was performed with a 100 degrees, 5-7.5 MHz convex probe on a Pentax endoscope and CT using the byphasic spiral technique. RESULTS: The 11 GIST cases were ranked according to the criteria of malignancy defined by each of the two imaging techniques. There was a close correlation between the imaging-based and pathological classifications. CONCLUSIONS: Size and homogeneous pattern prove to be the most reliable criteria to differentiate between benign and malignant lesions. EUS is more accurate than CT in diagnosing the nature of GIST, but CT allows a more complete and comprehensive evaluation. The data provided by the imaging techniques are fundamental to plan the therapeutic approach. PMID- 11859303 TI - Endorectal coil MRI in local staging of rectal cancer. AB - PURPOSE: The choice of the therapeutic strategies in patients affected with rectal cancer is strictly dependent by the tumor stage. So, in order to obtain an improvement in preoperative staging accuracy, new imaging modalities are now under investigation. The aim of this work is the evaluation of endorectal-coil MRI in the local staging of rectal cancer. MATERIAL AND METHODS: Fourty-three patients affected with histologically proven rectal cancer, have been evaluated by an high-field strength magnet (1.5 T). In 14/43 patients neoadjuvant pre operative chemotherapy had been previously performed. In all cases axial SE T1w and FSE T2w sequences and coronal or sagittal FSE T2w sequences, with and without fat suppression, were performed. Basing upon the TNM staging system and the previously reported MRI signs the local extent of the tumor was evaluated, focusing about the rectal wall infiltration and the perirectal lymph nodes involvement. All the patients underwent surgery and a comparative evaluation of MRI and pathological staging was done. RESULTS: At MRI the tumor was detected in 38/43 patients. In evaluating wall infiltration the MRI results agreed with pathological results in 89% of patients and showed 92% accuracy in T1-T2 stage and 94% in T3. In evaluating perirectal lymph nodes metastases MRI showed 69% accuracy, 82% sensitivity and 55%specificity. DISCUSSION AND CONCLUSIONS: The poor accuracy of CT and body-coil MRI in evaluating wall involvement in patients with rectal cancer is mainly related to their inability to demonstrate the single layers of the rectal wall. So transrectal ultrasound is now the first choice modalitiy in local staging of rectal cancer. However transrectal ultrasound showed low sensitivity in detecting perirectal lymph nodes metastases and low accuracy in evaluating the patients previously undergone to neoadjuvant chemotherapy or radiotherapy. On the other hand the improvement of MRI sequences and the availability of the endorectal coils allowed to visualize the single layers of the rectal wall so making the endorectal-coil MRI a reliable imaging technique to stage rectal cancer. The results of our work demonstrate a good diagnostic accuracy of endorectal-coil MRI in local staging of rectal cancer, in particular the degree of rectal wall infiltration was well demonstrated, while the perirectal lymph nodes metastases were demonstrated with less accuracy. The long examination time, the costs and the movement-related artefacts are the main limits of MRI. In particular the movement-related artifacts sometime do not allow the visualization of the wall layers so lowering the diagnostic accuracy in demonstrating the tumor wall infiltration. In conclusion, even though endorectal coil MRI proved to be a reliable imaging technique in local staging of rectal cancer, at present we are not able to state what may be its real role in diagnostic evaluation of the patients with rectal cancer, in particular if compared to endorectal ultrasound. Further, comparative studies, based upon larger patients series are probably needed to draw a definitive conclusion. PMID- 11859304 TI - Noncontrast CT assessment of early cerebral ischaemia. AB - PURPOSE: To assess the diagnostic value of early signs of ischaemic cerebral infarction detected by unenhanced CT in the first 6-10 hours. MATERIAL AND METHODS: We reviewed the CT examinations of 42 patients (mean age: 61 years, range: 35-79) with suspected ischaemic stroke. We assessed CT findings at 6-10 hours of the onset of stroke for hemilateral evidence of main cerebral artery hyperdensity, sulcal effacement, liquoral space asymmetry, hypodensity of grey matter. The CT scans were performed without contrast medium. RESULTS: The topographic pattern of cerebral infarctions was: middle cerebral artery territory in 25 patients, anterior cerebral in 9, striatal lacunar infarction in 2, posterior junctional infarction in 5, anterior junctional infarction in 1. Early signs of infarction were present in 24 patients (57%). CT scans showed early signs in 20 cases (80%) of middle cerebral infarctions; 8 (32%) had middle cerebral artery hyperdensity; 3 (12%) had middle cerebral artery hyperdensity and sulcal effacement; 4 (16%) had sulcal effacement; 2 (8%) had liquoral space asymmetry; 3 (12%) had hypodensity of grey matter and liquoral space asymmetry. CT scans showed early signs in 4/9 (44%) of anterior cerebral infarctions. Sensitivity and specificity of early CT to cerebral infarction was 57% and 100%. The three cases with both hyperdense middle cerebral artery and sulcal effacement died of transtentorial herniation within the 10th day. The seven other deaths occurred in patients without early signs or particular patterns appearing in subsequent CT. DISCUSSION: In the management of ischaemic stroke the aim of neuroradiologic methods is to provide exact direction to immediate therapy by early diagnosis. In such cases the use of CT scanning aims at detecting signs of two main alterations of infarction: vascular occlusion and brain oedema. Middle cerebral artery hyperdensity, showing steady correspondence to infarction site and frequent disappearance on the follow-up CT, is indicative of embolic occlusion. Signs of "mass effect" are evident from the early stages in relation to the substantial concomitance of various types of brain oedema. The semeiology discussed in this study is more clearly detectable in middle cerebral artery infarction because this territory is the main site of embolic occlusion, and its larger size increases the "mass effect" due to oedema. The sensitivity obtained in this study is among the lowest values reported, which likely relates to our choice to use short scan times. PMID- 11859305 TI - Cost analysis of equipment failure of a radiology department and possible choices about maintenance. AB - PURPOSE: Our aim was to evaluate the economic impact of equipment failures in a radiology department with a view to guiding maintenance policy decisions. MATERIAL AND METHODS: We assessed the negative economic impact caused by the interruption of activity of a radiodiagnostics section due to equipment failure, taking into account: the effects occurring during the first day of equipment down time (assuming that the equipment failure occurs in the middle of the shift) and the effects during the following days until the repair of the failure; the effects occurring in the short- and long-term. To exemplify the negative impact of inactivity due to equipment failure, we chose three radiology sections with different levels of technological and operational complexity (chest radiology, gastrointestinal radiology and remote-controlled diagnostics). For each, we evaluated the loss of contribution margin and the idle capacity costs (short- and long-term impact). RESULTS: The negative economic effects were: for thoracic radiology, 496,77 Euro in the first day, and 30,99 Euro from the second day onwards; for gastrointestinal radiology, 526,40 Euro for the first day, and 730,39 Euro from the second day onwards; for remote-controlled diagnostics, 786,25 Euro for the first day, and 927,67 Euro from the second days onwards. DISCUSSION: Our results indicate that the level of idle capacity costs (mainly equipment and staff) increases with the complexity of the equipment, whereas the contribution margin appears to fluctuate, because the charges are state-imposed and do not vary with the complexity of equipment. Moreover, our analysis shows that if the workload of a broken machine can easily be assigned to an additional shift using another machine, losses are considerably reduced from the second day onwards. Once the negative economic impact of equipment failures has been evaluated, the second step is to choose the best kind of maintenance. CONCLUSIONS: A sound calculation of the economic impact of equipment failures is very useful for guiding the head of department and the hospital manager in deciding whether to purchase maintenance services (or a long-term guarantee) from the equipment manufacturer, to set up an auxiliary centre for maintenance and repair, or to purchase a third-party maintenance contract. PMID- 11859306 TI - Total skin electron beam therapy in mycosis fungoides. Our experience from 1985 to 1999. AB - PURPOSE: The specific goal of this retrospective study is to evaluate the role of total skin electron beam therapy (TSEBT) in the treatment of Mycosis Fungoides (MF) and to assess the most significant prognostic factors in univariate and multivariate analyses. MATERIAL AND METHODS: From January 1985 to December 1999, 92 TSEBT (Stanford Standing technique) were performed on a total of 86 patients (63 with Mycosis Fungoides, 6 with Sezary Syndrome and 17 with Cutaneous Lymphomas). This study considers only the Mycosis Fungoides group, which consisted of 60 cases evaluable for response, survival and toxicity. The distribution of patients by stage (MFCG Staging Classification, 1991) was 21, 5, 12, 22 for stages I, II, III and IV, respectively. RESULTS: The overall response rate was 96.6% (58/60) with complete response (CR) in 50/60 patients (83.3%) and partial response (PR) in 8 cases (13.3%). The five-year and ten-year actuarial overall survival (OS) was 50% and 45%, respectively. Local control, intended as control of the disease in the skin, was 35% at five years and 20% at ten years, and was correlated with skin involvement. The prognostic factors confirmed by the multivariate analysis for both overall survival and local control were: T (p<0.001) and response after TSEBT (p<0.001). The treatment was very well tolerated. CONCLUSIONS: Our study showed good results in terms of response and survival with a long follow-up time (mean value 40 months). We confirm that TSEBT yields very good results in early-stage MF; additional trials of combined modality and investigational therapies are needed to improve the outcome for advanced-stage disease. PMID- 11859307 TI - Double array cochlear implant: CT findings. PMID- 11859309 TI - Giant sigmoid diverticulum. A rare manifestation of a common event. A case report. PMID- 11859308 TI - Adult pancreatoblastoma. A case report. PMID- 11859310 TI - Perirenal urinary extravasation: imaging and therapy. Report of two cases. PMID- 11859312 TI - Relax! No surgery. PMID- 11859311 TI - Extranodal Hodgkin lymphoma with renal, splenic and uterine involvement. A case report. PMID- 11859320 TI - Awards for endeavour. PMID- 11859313 TI - Imaging diagnosis of middle ear lesions. PMID- 11859321 TI - Putting politics back into science. PMID- 11859322 TI - Thoughts on (dis)credits. PMID- 11859324 TI - Biologists apprehensive over US moves to censor information flow. PMID- 11859325 TI - Protein chemists favour automatic answers. PMID- 11859328 TI - Live lung tissue enlisted in fight against tuberculosis. PMID- 11859329 TI - Earth-science centre targets core questions. PMID- 11859331 TI - Blast-off approaches for eagle-eyed orbiter. PMID- 11859334 TI - Life in the deep freeze. PMID- 11859335 TI - Canada: Northern poles of excellence. PMID- 11859337 TI - Sklyarov: big business vs academic freedom. PMID- 11859336 TI - Excitement over X-ray lasers is excessive. PMID- 11859338 TI - More light on pioneers of electrochemistry. PMID- 11859344 TI - Mindless mastery. PMID- 11859339 TI - Rank injustice. PMID- 11859346 TI - Extra dimension with X-rays. PMID- 11859345 TI - No more free lunch. PMID- 11859347 TI - Brouhaha over the other yeast. PMID- 11859349 TI - How insects lose their limbs. PMID- 11859351 TI - Max Perutz (1914-2002). PMID- 11859352 TI - Science of nuclear warheads. PMID- 11859353 TI - A cat cloned by nuclear transplantation. AB - Sheep, mice, cattle, goats and pigs have all been cloned by transfer of a donor cell nucleus into an enucleated ovum, and now we add the successful cloning of a cat (Felis domesticus) to this list. However, this cloning technology may not be readily extendable to other mammalian species if our understanding of their reproductive processes is limited or if there are species-specific obstacles. PMID- 11859359 TI - The role of the thermohaline circulation in abrupt climate change. AB - The possibility of a reduced Atlantic thermohaline circulation in response to increases in greenhouse-gas concentrations has been demonstrated in a number of simulations with general circulation models of the coupled ocean-atmosphere system. But it remains difficult to assess the likelihood of future changes in the thermohaline circulation, mainly owing to poorly constrained model parameterizations and uncertainties in the response of the climate system to greenhouse warming. Analyses of past abrupt climate changes help to solve these problems. Data and models both suggest that abrupt climate change during the last glaciation originated through changes in the Atlantic thermohaline circulation in response to small changes in the hydrological cycle. Atmospheric and oceanic responses to these changes were then transmitted globally through a number of feedbacks. The palaeoclimate data and the model results also indicate that the stability of the thermohaline circulation depends on the mean climate state. PMID- 11859361 TI - Diverse supernova sources of pre-solar material inferred from molybdenum isotopes in meteorites. AB - Variations in the isotopic composition of some components in primitive meteorites demonstrate that the pre-solar material was not completely homogenized, nor was it processed at sufficiently high temperatures to erase the signatures of the diverse stellar sources. This is in accord with the observation that accretion disks of young stellar objects are at relatively low temperatures. Carbonaceous chondrites are considered to represent the 'average' Solar System composition; the rare pre-solar grains in the matrixes of carbonaceous chondrites have been used to identify some sources of the pre-solar material. Here we report that the molybdenum isotopic composition of bulk carbonaceous chondrites is distinctly different from the accepted average solar value. We show that the Mo data require the presence of material produced in at least two different r-processes, and that the contribution from the p-process material is decoupled from the r-process, all occurring in supernova explosions. This is consistent with the emerging picture of diverse sources inferred from short-lived isotopes in the early Solar System and elemental analyses of metal-poor stars. PMID- 11859362 TI - Ultra-broadband semiconductor laser. AB - The fundamental mechanism behind laser action leads in general only to narrowband, single-wavelength emission. Several approaches for achieving spectrally broadband laser action have been put forward, such as enhancing the optical feedback in the wings of the gain spectrum, multi-peaked gain spectra, and the most favoured technique at present, ultrashort pulse excitation. Each of these approaches has drawbacks, such as a complex external laser cavity configuration, a non-flat optical gain envelope function, or an inability to operate in continuous mode, respectively. Here we present a monolithic, mid infrared 'supercontinuum' semiconductor laser that has none of these drawbacks. We adopt a quantum cascade configuration, where a number of dissimilar intersubband optical transitions are made to cooperate in order to provide broadband optical gain from 5 to 8 microm wavelength. Laser action with a Fabry Perot spectrum covering all wavelengths from 6 to 8 microm simultaneously is demonstrated with this approach. Lasers that emit light over such an extremely wide wavelength range are of interest for applications as varied as terabit optical data communications or ultra-precision metrology and spectroscopy. PMID- 11859363 TI - Three-dimensional X-ray structural microscopy with submicrometre resolution. AB - Advanced materials and processing techniques are based largely on the generation and control of non-homogeneous microstructures, such as precipitates and grain boundaries. X-ray tomography can provide three-dimensional density and chemical distributions of such structures with submicrometre resolution; structural methods exist that give submicrometre resolution in two dimensions; and techniques are available for obtaining grain-centroid positions and grain-average strains in three dimensions. But non-destructive point-to-point three-dimensional structural probes have not hitherto been available for investigations at the critical mesoscopic length scales (tenths to hundreds of micrometres). As a result, investigations of three-dimensional mesoscale phenomena--such as grain growth, deformation, crumpling and strain-gradient effects--rely increasingly on computation and modelling without direct experimental input. Here we describe a three-dimensional X-ray microscopy technique that uses polychromatic synchrotron X-ray microbeams to probe local crystal structure, orientation and strain tensors with submicrometre spatial resolution. We demonstrate the utility of this approach with micrometre-resolution three-dimensional measurements of grain orientations and sizes in polycrystalline aluminium, and with micrometre depth resolved measurements of elastic strain tensors in cylindrically bent silicon. This technique is applicable to single-crystal, polycrystalline, composite and functionally graded materials. PMID- 11859360 TI - The genome sequence of Schizosaccharomyces pombe. AB - We have sequenced and annotated the genome of fission yeast (Schizosaccharomyces pombe), which contains the smallest number of protein-coding genes yet recorded for a eukaryote: 4,824. The centromeres are between 35 and 110 kilobases (kb) and contain related repeats including a highly conserved 1.8-kb element. Regions upstream of genes are longer than in budding yeast (Saccharomyces cerevisiae), possibly reflecting more-extended control regions. Some 43% of the genes contain introns, of which there are 4,730. Fifty genes have significant similarity with human disease genes; half of these are cancer related. We identify highly conserved genes important for eukaryotic cell organization including those required for the cytoskeleton, compartmentation, cell-cycle control, proteolysis, protein phosphorylation and RNA splicing. These genes may have originated with the appearance of eukaryotic life. Few similarly conserved genes that are important for multicellular organization were identified, suggesting that the transition from prokaryotes to eukaryotes required more new genes than did the transition from unicellular to multicellular organization. PMID- 11859364 TI - Chiral recognition in dimerization of adsorbed cysteine observed by scanning tunnelling microscopy. AB - Stereochemistry plays a central role in controlling molecular recognition and interaction: the chemical and biological properties of molecules depend not only on the nature of their constituent atoms but also on how these atoms are positioned in space. Chiral specificity is consequently fundamental in chemical biology and pharmacology and has accordingly been widely studied. Advances in scanning probe microscopies now make it possible to probe chiral phenomena at surfaces at the molecular level. These methods have been used to determine the chirality of adsorbed molecules, and to provide direct evidence for chiral discrimination in molecular interactions and the spontaneous resolution of adsorbates into extended enantiomerically pure overlayers. Here we report scanning tunnelling microscopy studies of cysteine adsorbed to a (110) gold surface, which show that molecular pairs formed from a racemic mixture of this naturally occurring amino acid are exclusively homochiral, and that their binding to the gold surface is associated with local surface restructuring. Density functional theory calculations indicate that the chiral specificity of the dimer formation process is driven by the optimization of three bonds on each cysteine molecule. These findings thus provide a clear molecular-level illustration of the well known three-point contact model for chiral recognition in a simple bimolecular system. PMID- 11859365 TI - Deterioration of the seventeenth-century warship Vasa by internal formation of sulphuric acid. AB - The seventeenth-century Swedish warship, Vasa, was recovered in good condition after 333 years in the cold brackish water of Stockholm harbour. After extensive treatment to stabilize and dry the ship's timbers, the ship has been on display in the Vasa Museum since 1990. However, high acidity and a rapid spread of sulphate salts were recently observed on many wooden surfaces, which threaten the continued preservation of the Vasa. Here we show that, in addition to concentrations of sulphate mostly on the surface of oak beams, elemental sulphur has accumulated within the beams (0.2-4 per cent by mass), and also sulphur compounds of intermediate oxidation states exist. The overall quantity of elemental sulphur could produce up to 5,000 kg of sulphuric acid when fully oxidized. We suggest that the oxidation of the reduced sulphur--which probably originated from the penetration of hydrogen sulphide into the timbers as they were exposed to the anoxic water--is being catalysed by iron species released from the completely corroded original iron bolts, as well as from those inserted after salvage. Treatments to arrest acid wood hydrolysis of the Vasa and other wooden marine-archaeological artefacts should therefore focus on the removal of sulphur and iron compounds. PMID- 11859366 TI - Transient dynamics of vulcanian explosions and column collapse. AB - Several analytical and numerical eruption models have provided insight into volcanic eruption behaviour, but most address plinian-type eruptions where vent conditions are quasi-steady. Only a few studies have explored the physics of short-duration vulcanian explosions with unsteady vent conditions and blast events. Here we present a technique that links unsteady vent flux of vulcanian explosions to the resulting dispersal of volcanic ejecta, using a numerical, axisymmetric model with multiple particle sizes. We use observational data from well documented explosions in 1997 at the Soufriere Hills volcano in Montserrat, West Indies, to constrain pre-eruptive subsurface initial conditions and to compare with our simulation results. The resulting simulations duplicate many features of the observed explosions, showing transitional behaviour where mass is divided between a buoyant plume and hazardous radial pyroclastic currents fed by a collapsing fountain. We find that leakage of volcanic gas from the conduit through surrounding rocks over a short period (of the order of 10 hours) or retarded exsolution can dictate the style of explosion. Our simulations also reveal the internal plume dynamics and particle-size segregation mechanisms that may occur in such eruptions. PMID- 11859367 TI - Global environmental controls of diversity in large herbivores. AB - Large mammalian herbivores occupy half of the earth's land surface and are important both ecologically and economically, but their diversity is threatened by human activities. We investigated how the diversity of large herbivores changes across gradients of global precipitation and soil fertility. Here we show that more plant-available moisture reduces the nutrient content of plants but increases productivity, whereas more plant-available nutrients increase both of these factors. Because larger herbivore species tolerate lower plant nutrient content but require greater plant abundance, the highest potential herbivore diversity should occur in locations with intermediate moisture and high nutrients. These areas are dry enough to yield high quality plants and support smaller herbivores, but productive enough to support larger herbivores. These predictions fit with observed patterns of body size and diversity for large mammalian herbivores in North America, Africa and Australia, and yield a global map of regions with potentially high herbivore diversity. Thus, gradients of precipitation, temperature and soil fertility might explain the global distribution of large herbivore diversity and help to identify crucial areas for conservation and restoration. PMID- 11859368 TI - Climate change and the resurgence of malaria in the East African highlands. AB - The public health and economic consequences of Plasmodium falciparum malaria are once again regarded as priorities for global development. There has been much speculation on whether anthropogenic climate change is exacerbating the malaria problem, especially in areas of high altitude where P. falciparum transmission is limited by low temperature. The International Panel on Climate Change has concluded that there is likely to be a net extension in the distribution of malaria and an increase in incidence within this range. We investigated long-term meteorological trends in four high-altitude sites in East Africa, where increases in malaria have been reported in the past two decades. Here we show that temperature, rainfall, vapour pressure and the number of months suitable for P. falciparum transmission have not changed significantly during the past century or during the period of reported malaria resurgence. A high degree of temporal and spatial variation in the climate of East Africa suggests further that claimed associations between local malaria resurgences and regional changes in climate are overly simplistic. PMID- 11859369 TI - Evolution of a transcriptional repression domain in an insect Hox protein. AB - Homeotic (Hox) genes code for principal transcriptional regulators of animal body regionalization. The duplication and divergence of Hox genes, changes in their regulation, and changes in the regulation of Hox target genes have all been implicated in the evolution of animal diversity. It is not known whether Hox proteins have also acquired new activities during the evolution of specific lineages. Amino-acid sequences outside the DNA-binding homeodomains of Hox orthologues diverge significantly. These sequence differences may be neutral with respect to protein function, or they could be involved in the functional divergence of Hox proteins and the evolutionary diversification of animals. Here, we identify a transcriptional repression domain in the carboxy-terminal region of the Drosophila Ultrabithorax (Ubx) protein. This domain is highly conserved among Ubx orthologues in other insects, but is absent from Ubx in other arthropods and onychophorans. The evolution of this domain may have facilitated the greater morphological diversification of posterior thoracic and anterior abdominal segments characteristic of modern insects. PMID- 11859370 TI - Hox protein mutation and macroevolution of the insect body plan. AB - A fascinating question in biology is how molecular changes in developmental pathways lead to macroevolutionary changes in morphology. Mutations in homeotic (Hox) genes have long been suggested as potential causes of morphological evolution, and there is abundant evidence that some changes in Hox expression patterns correlate with transitions in animal axial pattern. A major morphological transition in metazoans occurred about 400 million years ago, when six-legged insects diverged from crustacean-like arthropod ancestors with multiple limbs. In Drosophila melanogaster and other insects, the Ultrabithorax (Ubx) and abdominal A (AbdA, also abd-A) Hox proteins are expressed largely in the abdominal segments, where they can suppress thoracic leg development during embryogenesis. In a branchiopod crustacean, Ubx/AbdA proteins are expressed in both thorax and abdomen, including the limb primordia, but do not repress limbs. Previous studies led us to propose that gain and loss of transcriptional activation and repression functions in Hox proteins was a plausible mechanism to diversify morphology during animal evolution. Here we show that naturally selected alteration of the Ubx protein is linked to the evolutionary transition to hexapod limb pattern. PMID- 11859371 TI - Numerical representation for action in the parietal cortex of the monkey. AB - The anterior part of the parietal association area in the cerebral cortex of primates has been implicated in the integration of somatosensory signals, which generate neural images of body parts and apposed objects and provide signals for sensorial guidance of movements. Here we show that this area is active in primates performing numerically based behavioural tasks. We required monkeys to select and perform movement A five times, switch to movement B for five repetitions, and return to movement A, in a cyclical fashion. Cellular activity in the superior parietal lobule reflected the number of self-movement executions. For the most part, the number-selective activity was also specific for the type of movement. This type of numerical representation of self-action was seen less often in the inferior parietal lobule, and rarely in the primary somatosensory cortex. Such activity in the superior parietal lobule is useful for processing numerical information, which is necessary to provide a foundation for the forthcoming motor selection. PMID- 11859372 TI - BH3-only Bcl-2 family member Bim is required for apoptosis of autoreactive thymocytes. AB - During lymphocyte development, the assembly of genes coding for antigen receptors occurs by the combinatorial linking of gene segments. The stochastic nature of this process gives rise to lymphocytes that can recognize self-antigens, thereby having the potential to induce autoimmune disease. Such autoreactive lymphocytes can be silenced by developmental arrest or unresponsiveness (anergy), or can be deleted from the repertoire by cell death. In the thymus, developing T lymphocytes (thymocytes) bearing a T-cell receptor (TCR)-CD3 complex that engages self-antigens are induced to undergo programmed cell death (apoptosis), but the mechanisms ensuring this 'negative selection' are unclear. We now report that thymocytes lacking the pro-apoptotic Bcl-2 family member Bim (also known as Bcl2l11) are refractory to apoptosis induced by TCR-CD3 stimulation. Moreover, in transgenic mice expressing autoreactive TCRs that provoke widespread deletion, Bim deficiency severely impaired thymocyte killing. TCR ligation upregulated Bim expression and promoted interaction of Bim with Bcl-XL, inhibiting its survival function. These findings identify Bim as an essential initiator of apoptosis in thymocyte-negative selection. PMID- 11859373 TI - De novo pyrimidine biosynthesis is required for virulence of Toxoplasma gondii. AB - Toxoplasma gondii is a ubiquitous protozoan parasite that is responsible for severe congenital birth defects and fatal toxoplasmic encephalitis in immunocompromized people. Fundamental aspects of obligate intracellular replication and pathogenesis are only now beginning to emerge for protozoan parasites. T. gondii has a fragmented pathway for salvaging pyrimidine nucleobases derived from the parasite or host cell, and this limited pyrimidine salvage capacity is funnelled exclusively through uracil phosphoribosyltransferase. Disrupting the function of this enzyme does not affect the growth of T. gondii tachyzoites, which suggests that the de novo pyrimidine biosynthesis pathway may be necessary for growth. We have examined the virulence of T. gondii mutants that lack carbamoyl phosphate synthetase II (uracil auxotrophs) to determine whether de novo pyrimidine biosynthesis is required in vivo. Here we show that T. gondii uracil auxotrophs are completely avirulent not only in immune-competent BALB/c mice but also in mice that lack interferon-gamma. A single injection of the uracil auxotroph into BALB/c mice induces long-term protective immunity to toxoplasmosis. Our findings indicate the significance of the de novo pyrimidine biosynthesis pathway for the virulence of parasitic protozoa, and suggest routes for developing vaccines and chemotherapy. PMID- 11859374 TI - A Rad26-Def1 complex coordinates repair and RNA pol II proteolysis in response to DNA damage. AB - Eukaryotic cells use multiple, highly conserved mechanisms to contend with ultraviolet-light-induced DNA damage. One important response mechanism is transcription-coupled repair (TCR), during which DNA lesions in the transcribed strand of an active gene are repaired much faster than in the genome overall. In mammalian cells, defective TCR gives rise to the severe human disorder Cockayne's syndrome (CS). The best-studied CS gene, CSB, codes for a Swi/Snf-like DNA dependent ATPase, whose yeast homologue is called Rad26 (ref. 4). Here we identify a yeast protein, termed Def1, which forms a complex with Rad26 in chromatin. The phenotypes of cells lacking DEF1 are consistent with a role for this factor in the DNA damage response, but Def1 is not required for TCR. Rather, def1 cells are compromised for transcript elongation, and are unable to degrade RNA polymerase II (RNAPII) in response to DNA damage. Our data suggest that RNAPII stalled at a DNA lesion triggers a coordinated rescue mechanism that requires the Rad26-Def1 complex, and that Def1 enables ubiquitination and proteolysis of RNAPII when the lesion cannot be rapidly removed by Rad26-promoted DNA repair. PMID- 11859375 TI - Structural basis for acidic-cluster-dileucine sorting-signal recognition by VHS domains. AB - Specific sorting signals direct transmembrane proteins to the compartments of the endosomal-lysosomal system. Acidic-cluster-dileucine signals present within the cytoplasmic tails of sorting receptors, such as the cation-independent and cation dependent mannose-6-phosphate receptors, are recognized by the GGA (Golgi localized, gamma-ear-containing, ADP-ribosylation-factor-binding) proteins. The VHS (Vps27p, Hrs and STAM) domains of the GGA proteins are responsible for the highly specific recognition of these acidic-cluster-dileucine signals. Here we report the structures of the VHS domain of human GGA3 complexed with signals from both mannose-6-phosphate receptors. The signals bind in an extended conformation to helices 6 and 8 of the VHS domain. The structures highlight an Asp residue separated by two residues from a dileucine sequence as critical recognition elements. The side chains of the Asp-X-X-Leu-Leu sequence interact with subsites consisting of one electropositive and two shallow hydrophobic pockets, respectively. The rigid spatial alignment of the three binding subsites leads to high specificity. PMID- 11859378 TI - Revisiting Adverse Drug Reactions. PMID- 11859376 TI - Structural basis for recognition of acidic-cluster dileucine sequence by GGA1. AB - GGAs (Golgi-localizing, gamma-adaptin ear homology domain, ARF-interacting proteins) are critical for the transport of soluble proteins from the trans-Golgi network (TGN) to endosomes/lysosomes by means of interactions with TGN-sorting receptors, ADP-ribosylation factor (ARF), and clathrin. The amino-terminal VHS domains of GGAs form complexes with the cytoplasmic domains of sorting receptors by recognizing acidic-cluster dileucine (ACLL) sequences. Here we report the X ray structure of the GGA1 VHS domain alone, and in complex with the carboxy terminal peptide of cation-independent mannose 6-phosphate receptor containing an ACLL sequence. The VHS domain forms a super helix with eight alpha-helices, similar to the VHS domains of TOM1 and Hrs. Unidirectional movements of helices alpha6 and alpha8, and some of their side chains, create a set of electrostatic and hydrophobic interactions for correct recognition of the ACLL peptide. This recognition mechanism provides the basis for regulation of protein transport from the TGN to endosomes/lysosomes, which is shared by sortilin and low-density lipoprotein receptor-related protein. PMID- 11859379 TI - Evidence of an Eicosanoid Contribution to IL-1 Induction of IL-6 in Human Articular Chondrocytes. AB - It has been demonstrated previously that interleukin-1 (IL-1) induces articular cartilage explants and chondrocytes in culture to produce elevated levels of inflammatory mediators such as interleukin-6 (IL-6) and prostaglandins. Previous studies have also demonstrated a relationship between IL-6 secretion and the ability of IL-1 to modulate proteoglycan synthesis by chondrocytes. In this study we have utilized an alginate culture system in an effort to investigate a role for eicosanoids in IL-1 induction of IL-6 expression in human articular chondrocytes. IL-1 treatment of chondrocytes cultured in alginate resulted in increased synthesis of IL-6 and prostaglandins, but not leukotrienes. Cyclo oxygenase inhibitor, indomethacin (5 &mgr;g ml(minus sign1)), was able to inhibit prostaglandin synthesis to below basal levels with no significant effect on the levels of IL-6 released by chondrocytes in response to IL-1. When chondrocytes were treated with 5 &mgr;g ml(minus sign1) indomethacin and 10 &mgr;M of the general lipoxygenase inhibitor, nordihydroguiaretic acid (NDGA), an approximate 50% decrease in IL-1-induced IL-6 expression was observed. Alone, levels of NDGA specific for lipoxygenase inhibition (10 &mgr;M) did not affect IL-1-induced IL-6 expression, but higher levels of NDGA (50 &mgr;M) which inhibited both prostaglandin and leukotriene biosynthesis reduced IL-1-induced IL-6 expression to the same extent as that observed with 5 &mgr;g ml(minus sign1) indomethacin and 10 &mgr;M NDGA. This inhibition of IL-6 expression by NDGA and indomethacin was dose responsive and also reversible with the addition of exogenous prostaglandin E(2) (PGE(2)) or leukotriene B(4) (LTB(4)). Although IL-1-induced IL-6 expression was only affected when both prostaglandin and leukotriene biosynthesis were inhibited, elevated levels of PGE(2) but not leukotriene B(4), C(4), D(4), or E(4) were observed in the culture medium of IL-1-treated chondrocytes. These findings may indicate that cyclo-oxygenase products such as PGE(2) normally contribute to IL-1 induction of IL-6 expression in chondrocytes, and under conditions when cyclo-oxygenase is inhibited, lipoxygenase products alternatively contribute to this response. PMID- 11859380 TI - T-Cell Derived Lymphokines as Regulators of Chronic Inflammation: Potential Targets for Immunomodulation? AB - In recent years, compelling evidence has accumulated that inflammation results from a cascade of events that are orchestrated by cytokines. Cytokines are products of nonimmune and immune cells which regulate the recruitment, differentiation, and proliferation of inflammatory cells. Although there is redundancy in cytokine production and function, major pathways of cytokines have been identified. Besides macrophages, T cells represent important sources of pro- and anti-inflammatory cytokines. In support of a critical regulatory role of T cells in inflammation, two T cell subtypes characterized by different cytokine profiles have been described. TH1 cells produce interleukin-2 (IL-2) and interferon-gamma (IFN-gamma) and are prone to interact with macrophages. TH2 cells are producers of IL-4, IL-5, and IL-10 and are capable of supporting B cells and eosinophils. Here, we are providing evidence that different human inflammatory diseases are characterized by distinct in situ cytokine patterns. Giant cell vasculitis represents a typical TH1-like disease, whereas TH2 helper cells appear to be more important in rheumatoid arthritis. Given the association of different diseases with cytokine profiles, the question arises how the microenvironment influences cytokine pattern and thus disease and whether mediators produced at the site of inflammation could serve as therapeutic targets to modulate the cytokine cascade. Prostaglandin E(2) (PGE(2)) is an important inflammatory mediator. We have examined the influence of PGE(2) and the PGE analog misoprostol on T-cell function and T-cell cytokine production. Here, we describe that IL-2 and IFN-gamma production are sensitive to PGE(2) and misoprostol, whereas IL-4 and IL-5 are unaffected. Thus, in the presence of PGE(2), TH0 cells acquire a TH2-like function, whereas TH1-like aspects are suppressed. We suggest that PGE(2) and misoprostol might represent useful modulators of the cytokine network. PMID- 11859381 TI - Modulation of the Expression of Glucocorticoid Receptors in Synovial Fibroblasts and Chondrocytes by Prostaglandins and NSAIDs. AB - Endogenous glucocorticoids are of prime importance in the maintenance of cellular homeostasis through their binding to specific cell receptors. Under normal conditions, physiological concentrations of cortisol are potentially capable of suppressing metalloprotease synthesis by chondrocytes and synoviocytes. This hormone action is like to represent one of the major pathways through which the catabolism of cartilage under physiological conditions is kept in a homeostatic state. In osteoarthritis (OA), a severe reduction (about 50%) in the glucocorticoid receptor (GR) level in chondrocytes was found to be sufficient to alter the cellular steroid resistance in terms of the synthesis of destructive enzymes. Prostaglandins of the E series and dibutyryl cyclic AMP increased the GR level in normal and OA human articular chondrocytes. Synthetic PGE(1) (misoprostol) also induced upregulation of GR expression in chondrocytes. Naproxen and indomethacin, but not tiaprofenic acid, at therapeutic concentrations, significantly reduced the level of GR in synovial cells. This effect could be reversed by the additionl of misoprostol. These findings bring insight into the differential effects of nonsteroidal anti-inflammatory drugs (NSAIDs) on the GR system and may provide an explanation for the reduced level of GR found in OA chondrocytes. The addition of synthetic prostaglandins may prove to be of therapeutic importance in the treatment of arthritic diseases by potentiating the effects of therapeutically administered glucocorticoid on the reducation of the catabolism of articular cartilage. PMID- 11859382 TI - Forskolin Stimulates Aggrecan Gene Expression in Cultured Bovine Chondrocytes. AB - Prostaglandins are autacoids that elevate intracellular 3prime prime or minute:5prime prime or minute-cyclic adenosine monophosphate (cAMP) levels in chondrocytes and other cells in culture. To facilitate intracellular cAMP accumulation, bovine chondrocytes were incubated with forskolin alone or forskolin and isobutylmethylxanthine. Both significantly increased proteoglycan synthesis, which was inhibited by the cAMP-dependent protein kinase inhibitor H89. Sodium dodecyl sulfate/polyacrylamide gel electrophoresis (SDS/PAGE) on 3- 16% gels revealed the presence of two large proteoglycan core proteins which migrated more slowly than the 200-kDa marker protein and two small proteoglycan core proteins which migrated slightly slower than the 46-kDa marker. Northern blot hybridization, employing (32)P-labeled cDNA probes, showed that aggrecan steady-state mRNA levels were increased by forskolin and isobutylmethylxanthine after 1 h and 5 h incubation. Decorin and type II collagen mRNA levels were not altered under these conditions. Link protein mRNA levels were slightly elevated, but only at the 5-h time point. These results indicated that stimulation of intracellular cAMP accumulation by forskolin or forskolin and isobutylmethylxanthine resulted in augmented proteoglycan synthesis via increased steady-state aggrecan mRNA levels. Suppression of proteoglycan synthesis by the cAMP-dependent protein kinase inhibitor H89 suggested that cAMP-dependent protein kinase may also play a role in regulating the synthesis and completion of newly synthesized proteoglycans. PMID- 11859383 TI - Molecular Mechanisms Used in the Regulation of Aggrecan and Link Protein Synthesis by Chondrocytes. PMID- 11859384 TI - Effects of Misoprostol on Bone Resorption and Formation in Organ Culture. AB - Prostaglandins are potent stimulators of bone resorption and formation. Because misoprostol is an analog of prostaglandin E(1) (PGE(1)), we have examined its effects on resorption and formation in organ culture. The results were compared with PGE(2) which can stimulate resorption and both stimulate and inhibit bone formation. Resorption, measured as the release of previously incorporated (45)Ca from 5-day cultures of 19-day fetal-rat long bones, was increased by misoprostol 1.5-fold at 10(minus sign6) M and twofold at 10(minus sign5) M. The effect at 10(minus sign6) M was abolished by addition of indomethacin (10(minus sign6) M). PGE(2) was approximately 100 times more potent and was not affected by indomethacin in this system. In 21-day fetal-rat calvariae, cultured for 24 h in the presence of cortisol (10(minus sign7) M), misoprostol produced a dose-related increase in TdR incorporation between 10(minus sign5) and 10(minus sign7) M. PGE(2) appeared to be only 10-fold more potent in this response. The effects of misoprostol on incorporation of [(3)H]proline into collagenase digestible protein (CDP) and noncollagen protein (NCP) were measured in 72-h cultures, either with continuous treatment or 24-h treatment followed by 48 h in control medium. With continuous treatment at 10(minus sign6) M, misoprostol increased labeling of CDP twofold. A similar effect was observed with 24 h of treatment at 10(minus sign5) M, followed by 48 h in control medium. Again, PGE(2) was approximately 10-fold more potent than misoprostol. When calvariae were treated with insulin-like growth factor I, which increases CDP labeling by 2.5-fold, the effects of misoprostol and PGE(2) were inhibitory. We conclude that misoprostol resembles PGE(2) in its effects on bone but is less potent. Moreover, misoprostol may be relatively less effective in stimulating resorption than in stimulating formation. Therefore, an increase in bone turnover, possibly with a net anabolic effect, might occur in vivo with long-term misoprostol treatment. Misoprostol effects on bone turnover in humans could be evaluated using the sensitive biochemical markers for bone resorption and formation which are currently available. PMID- 11859385 TI - Prostaglandins and the Zone of Calcified Cartilage in Osteoarthritis. AB - The zone of calcified cartilage (ZCC) which provides the critical interface between cartilage and bone acts as the growth plate in the developing joint. In osteoarthritis, it has been hypothesized that the ZCC may again function in joint remodeling. This could result in thinning of the cartilage. This report is the first experimental confirmation of this hypothesis. Osteoarthritis was induced using the Hulth procedure in 2.3--2.7-kg rabbits. Approximately 3 weeks after surgery, half of the menisectomy rabbits and half of the nonmenisectomy rabbits were given 20 &mgr;g of misoprostol interarticularly for 5 days per week for 2.5 weeks. In the patellae and tibial plateau, the rate of movement of the tidemark of the ZCC was measured. Medial aspect femoral condyle cartilage was incubated in the presence of [(35)S]sulfate and [(3)H]proline. After menisectomy, rate of movement of the ZCC was dramatically increased and was unchanged by misoprostol. Proteoglycan synthesis was highly elevated in the osteoarthritis-induced knees, and misoprostol suppressed the rate of [(35)S]sulfate but not [(3)H]proline incorporation. PMID- 11859386 TI - The Cytochrome P450 Enzymes of Hepatic Drug Metabolism: How are their Activities Assessed In Vivo, and what is their Clinical Relevance? AB - The cytochromes P450 comprise a superfamily of mixed function oxidases responsible for the oxidation of numerous endobiotics and thousands of xenobiotics. Though they are distributed in many organ systems, the hepatic cytochromes P450 are most prominent in both the detoxification and bioactivation of xenobiotics. They have been investigated intensively for over 30 years; however a systematic nomenclature did not evolve until it became clear that numerous isoforms existed. Based on similarities of amino acid sequences, the cytochrome P450 (CYP) enzymes have now been classified by family, subfamily, and isoform. As expression systems have been developed to assist in the determination of which isoforms mediate the oxidations of which substrates, it has become possible to model binding sites for some of the isoforms. This enterprise has been assisted by the determination of the amino acide sequence and the X-ray crystal structure of the bacterial cytochrome P450, P450(CAM). As the CYP enzymes play such a vital role in drug metabolism, and as the activities of various CYP isoforms have been implicated in cancer risk, there is a compelling need to develop methods for genotyping or phenotyping subpopulations with regard to CYP activity. It should be especially important to phenotype CYP activities in subpopulations anytime CYP enzyme activity can be regulated by environmental factors. A priori knowledge about CYP activity in various subpopulations (e.g., the aged, those with hepatic dysfunction, women, those occupationally exposed to certain environmental chemicals) will be helpful in predicting therapeutic outcomes for drugs whose elimination is CYP dependent. Additionally, cancer risk has been shown to correlate with various CYP activities, suggesting that a priori knowledge about select CYP activities may be useful in cancer risk assessment. Several noninvasive and minimally invasive strategies for phenotyping CYP activities have emerged in recent years. These procedures hold the promise of permitting investigators to phenotype select CYP isoform or subfamily activities within defined subpopulations through the use of endobiotic probes of CYP activity, xenobiotic probes of CYP activity, and noninvasive and minimally invasive measurements of those probes. This article reviews the CYP enzymes, the minimally invasive and noninvasive methods for characterizing their hepatic activities in humans in vivo, and the significance of being able to phenotype their activities in vivo. PMID- 11859387 TI - Topical metronidazole for rosacea. AB - Rosacea is relatively common, typically occurring in individuals of Northern European and Celtic origin between 30 and 50 years of age. It is more common in women, but may be more severe in men. Currently there is no cure available for rosacea, but it can be controlled with topical and oral drug therapy. Topical metronidazole 1% cream is approved by the US FDA for the treatment of inflammatory lesions (papules and pustules) and erythema associated with rosacea. This treatment option is effective, safe and well tolerated. PMID- 11859388 TI - Treatments of choice for bullous pemphigoid. AB - Bullous pemphigoid (BP) is the most frequently occurring autoimmune blistering disease in Europe and North America. Although it is primarily a disease of the elderly, children and young adults can also develop it. The aim of treatment is to suppress the clinical signs and symptoms of BP without over-treating the patient, because BP tends to spontaneously remit in most patients within approximately 5 years. Mild or localized disease may respond to super-potent topical corticosteroids alone or in combination with tetracyclines with or without niacinamide. More severe or generalized disease usually requires systemic treatment with prednisolone (dose range from 20-70 mg/d). Additional immunosuppressant therapy is necessary for more refractory disease. PMID- 11859389 TI - Should involved-field radiation therapy be used as an adjunct to lymphoma autotransplantation? AB - Relapse at sites of prior disease involvement accounts for the majority of treatment failures following high-dose therapy and autologous transplantation for both Hodgkin's disease and non-Hodgkin's lymphoma. Several studies have demonstrated the utility of 'involved-field' radiation as a treatment modality in this setting to minimize disease bulk prior to transplants, to reduce relapse rates at sites of prior disease involvement and to improve local control for disease resistant to high-dose therapy. Other studies recommend caution due to potential toxicities including radiation-induced pneumonitis and secondary myelodysplasia. Further investigations are needed to better define the optimal extent, dose and timing of radiation in the setting of transplantation, as well as to identify those subsets of patients likely to be at a higher risk of radiation-induced morbidity. PMID- 11859390 TI - Second EBMT Workshop on reduced intensity allogeneic hemopoietic stem cell transplants (RI-HSCT). AB - A second meeting on reduced intensity allogeneic stem cell transplants (RI-HSCT) was convened in Zurich in February 2001 and focused on transplant-related mortality (TRM) and graft-versus-host disease (GVHD). Retrospective and prospective studies from the EBMT, national groups and single institutions included over 900 patients: the incidence of acute GVHD grade III-IV was 12% (1 17%), extensive chronic GVHD 42% (25-51%) and TRM 20% (14-38%). Conditioning regimens could be classified into four major groups based on (1) total body irradiation (TBI) 200 cGy, (2) busulfan 8 mg/kg, (3) thiotepa 10 mg/kg, and (4) melphalan 140 mg/m(2): most of these regimens are given in association with fludarabine in different doses and use mobilized peripheral blood as a source of stem cells. The incidence of TRM is similar if not identical for all four regimens, whereas the risk of acute GVHD and chronic GVHD may vary with different protocols. Reduced intensity transplant programs are being explored in patients above the age of 60 and in patients with solid tumors: encouraging results are being recorded in individual patients. Overall these data confirm that allogeneic HSCT can be performed in elderly patients, although a TRM of approximately 15% must be expected and is age dependent. A high rate of extensive chronic GVHD is seen and should be followed carefully. The term mini- or micro-transplant is probably misleading and one should refer to this procedure as an allogeneic HSCT and further classify the intensity of the conditioning regimen. PMID- 11859391 TI - Transient hematopoietic stem cell rescue using umbilical cord blood for a lethally irradiated nuclear accident victim. AB - We performed stem cell rescue and allogeneic skin transplantation on a lethally neutron-irradiated nuclear accident victim. HLA-DRB1 mismatched unrelated umbilical cord blood cells (2.08 x 10(7)/kg recipient body weight) were transplanted to an 8-10 Gy equivalent neutron-irradiated patient because of a lack of a suitable bone marrow or peripheral blood donor. Pre-transplant conditioning consisted of anti-thymocyte gamma-globulin alone, and GVHD prophylaxis was a combination of cyclosporine (CYA) and methylprednisolone (mPSL). Granulocyte colony-stimulating factor (G-CSF), erythropoietin (EPO), and thrombopoietin (TPO) were concurrently administered after transplantation. The absolute neutrophil count reached 0.5 x 10(9)/l on day 15, the reticulocyte count rose above 1% on day 23, and the platelet count was over 50 x 10(9)/l on day 27, respectively. Cytogenetic studies of blood and marrow showed donor/recipient mixed chimerism. Rapid autologous hematopoietic recovery was recognized after withdrawal of CYA and mPSL. Repeated pathological examinations of the skin revealed no evidence of acute GVHD. Eighty-two days after the irradiation, skin transplantation was performed to treat radiation burns. Almost 90% of the transplanted skin engrafted. Immunological examination after autologous hematopoietic recovery revealed an almost normal T cell count. However, immune functions were severely impaired. The patient died from infectious complication 210 days after the accident. PMID- 11859392 TI - CsA-based post-graft immunosuppression: the main factor for improving outcome of allografted patients with acquired aplastic anemia. A retrospective survey by the Spanish Group of Hematopoietic Transplantation. AB - A retrospective multicenter study was performed to assess the clinical results in patients with acquired aplastic anemia (AA) allografted over a 19 year period and to identify prognostic factors influencing survival. From April 1978 to December 1997, 176 patients were transplanted. Records from 160 receiving related matched bone marrow transplantation (BMT) were reviewed. Fifty-two percent of the patients were older than 20 years, 5% older than 40; 6.3% were untransfused at BMT and 56.2% had received prior treatments. Conditioning regimens were with chemotherapy in 43.7% of the procedures and with additional irradiation in 56.3%. Graft-versus-host disease (GVHD) prophylaxis was based on cyclosporin A (CsA) in 58.1% of the patients while methotrexate (MTX) was administered to 41.9%. Transplantation earlier on, a longer interval from diagnosis to BMT, GVHD prophylaxis with MTX, graft failure/rejection and acute severe GVHD were adverse factors for survival. The use of CsA emerged as the main factor for the improvement, inducing a significant decrease in graft failure/rejection rate and severe acute GVHD when compared with MTX alone. Radiation-containing regimens decreased the graft failure/rejection rate without improving survival due to the increased risk of acute GVHD. Age and number of transfusions pretransplant did not influence outcome. Survival achieved since 1991 is 79.79%, and graft failure and acute severe GVHD rates are 6.0% and 11.8%, respectively. In conclusion, CsA based post-graft immunosuppression has been crucial in achieving improved survival in patients with acquired AA up to 40 years of age. Regardless of CsA use, further improvement in survival was apparent with time, probably due to better skills in patient care. PMID- 11859393 TI - Idiotype protein-pulsed dendritic cells produce strong anti-myeloma effects after syngeneic stem cell transplantation in mice. AB - Dendritic cell (DC) vaccination represents an interesting immunotherapeutic option in the treatment of several malignancies. In multiple myeloma (MM) patients, vaccination with autologous idiotype (Id) protein-pulsed DC is feasible, but their antitumoral effectiveness was rather limited. To improve the therapeutic potential of DC therapy, we studied the immunological effects of syngeneic peripheral blood stem cell transplantation (PBSCT) given in conjunction with Id-loaded DC. Balb/c mice were inoculated i.p. with 5 x 10(5) of HOPC myeloma cells (Balb/c origin). Animals were immunized with three injections of 5 x 10(5) DC pulsed with the IgG2a(HOPC) or with a control immunoglobulin (Ig(control)). Some experimental groups of myeloma-bearing animals received total body irradiation (7.5 Gy) and a subsequent transplant of 2 x 10(7)syngeneic peripheral blood progenitor cells (PBPC) followed by DC therapy beginning at day 10 post transplant. Animals receiving DC therapy or syngeneic PBPCT alone neither induce long-term survival nor tumor-specific CTL reactivity in vitro. In marked contrast, combination of syngeneic PBPC transplantation and subsequent DC therapy resulted in 78% survival after a follow-up of 180 days. In addition, this treatment modality conferred a generation of Id peptide-specific CD8-mediated T cell reactivity. These data provide a rationale for DC-based vaccination in multiple myeloma patients administered post syngeneic transplantation. PMID- 11859394 TI - Chronic intestinal graft-versus-host disease: clinical, histological and immunohistochemical analysis of 17 children. AB - Graft-versus-host disease (GVHD) can be acute or chronic. The pathogenesis of chronic GVHD is unclear. Chronic GVHD affects mainly skin, liver and digestive tract. Intestinal involvement is uncommon and histological features are poorly described. We report here the clinical, histological and immunohistochemical features of chronic GVHD with intestinal involvement. Intestinal biopsies from children with chronic GVHD (n=17) were compared to control children (n=21: 10 non transplant cases, four non-GVHD transplant cases, seven acute GVHD). We evaluated clinical outcome, histological features and characterized immunohistochemically the immune cells involved locally. Chronic GVHD with intestinal involvement was usually multisystemic (88.2%) and preceded by acute GVHD in 88.2% of cases. The outcome was severe with complete recovery in only 58.8% of cases, and death related to chronic GVHD in 17.6% of cases. Histological features were characterized by (1) villous atrophy and (2) glandular lesions, mainly apoptotic with variable intensity and (3) lamina propria infiltrate with cytotoxic T lymphocytes (CD3+, CD8+, TiA1+, granzyme B-) which were significantly (P<0.001) increased compared to non-GVHD transplant and non-transplant controls. Therefore in chronic intestinal GVHD, the apoptotic process could be related to cytotoxic T lymphocytes. PMID- 11859395 TI - Weight loss and malnutrition in patients with chronic graft-versus-host disease. AB - It is well known that weight loss occurs in patients with chronic graft-versus host disease. However, the severity and frequency of weight loss in this population have not been adequately described. Recent data suggest that a body mass index (BMI) below 21.9 is an independent risk factor for mortality. In our analysis we have shown that out of 93 patients with cGVHD, 43% are malnourished as evidenced by a BMI less than 21.9 and 14% are severely malnourished (BMI less than 18.5). In addition, there is a clear trend showing that patients with active, ongoing cGVHD have lower BMIs (P = 0.02). Furthermore, we show that many symptoms thought to contribute to weight loss in patients with cGVHD, such as odynophagia and oral sensitivity, are not related to weight loss in our population. We conclude that, in all likelihood, unknown causes still exist that are responsible for weight loss in this group of patients. Elevated resting energy expenditure and elevated serum tumor necrosis factor-alpha are potential contributors to weight loss that will be tested in future studies. We also conclude that treating cGVHD aggressively may help reverse weight loss and malnutrition, which may be independent risk factors for mortality in this population. PMID- 11859397 TI - Comparison of short tandem repeat and variable number tandem repeat genetic markers for quantitative determination of allogeneic bone marrow transplant engraftment. AB - Variable number tandem repeats (VNTRs) were among the first genetic markers used to quantitate bone marrow transplant engraftment. The limitations of PCR-based VNTR markers in distinguishing some donor/recipient pairs has shown the need for additional genetic markers to analyze engraftment. Short tandem repeats (STRs) provide an excellent tool for this purpose because of their high degree of polymorphism and relatively short length. We compared STR analysis results with previous VNTR results for 16 post-transplantation samples from four allogeneic bone marrow transplant patients. Previously analyzed patient samples were chosen to cover the full range of engraftment. DNA samples from each patient were analyzed in a blinded fashion. Good quantitative correlation was found between STR and VNTR results in samples from all four patients. STR markers were informative in one patient for whom PCR-based VNTR markers were not available. Correlation of VNTR and STR methods helps to validate the use of STRs for the quantitative analysis of bone marrow transplant engraftment. This study demonstrates that STR-based human identity testing kits are well suited for engraftment analysis. PMID- 11859396 TI - Suspected delayed immune recovery against cytomegalovirus after reduced-intensity stem cell transplantation using anti-thymocyte globulin. AB - A reduced-intensity hematopoietic stem cell transplantation (RIST) regimen was developed to induce immunosuppression to facilitate the engraftment of donor cells. However, there have been concerns that the incidence of opportunistic infection may increase after this procedure. To address this problem, we retrospectively analyzed the medical records of 24 RIST recipients who were treated over a recent 16-month period for comparison with 31 recipients of conventional allogeneic transplantation (CST). The RIST regimen consisted of cladribine (0.66 mg/kg), busulfan (8 mg/kg), and rabbit anti-thymocyte globulin (ATG; 5-10 mg/kg). All of the patients received allogeneic peripheral blood stem cells from an HLA-identical or one-locus mismatched related donor. Although the incidence of positive CMV antigenemia was comparable between the two groups (58% vs 68%), RIST patients developed positive antigenemia significantly sooner than did CST patients (P = 0.01) and showed higher initial and maximum antigenemia values (P = 0.026 and P = 0.003, respectively). These findings may suggest that immune recovery against CMV was delayed after our RIST procedure, but this did not directly translate into an increase in clinically significant CMV disease. Early therapeutic intervention with ganciclovir might play a role in preventing the progression of early CMV infection to CMV disease. PMID- 11859398 TI - Extensive early apoptosis in frozen-thawed CD34-positive stem cells decreases threshold doses for haematological recovery after autologous peripheral blood progenitor cell transplantation. AB - Stem cell doses necessary for engraftment after myelo-ablative therapy as defined for fresh transplants vary largely. Loss of CD34+ cell quality after cryopreservation might contribute to this variation. With a new early apoptosis assay including the vital stain Syto16, together with the permeability marker 7 AAD, CD34+ cell viability in leucapheresis samples of 49 lymphoma patients receiving a BEAM regimen was analysed. After freeze-thawing large numbers of non viable, early apoptotic cells appeared, leading to only 42% viability compared to 72% using 7-AAD only. Based on this Syto16 staining in the frozen-thawed grafts, threshold numbers for adequate haematological recovery of 2.8-3.0 x 10(6) CD34+ cells/kg body weight determined for fresh grafts, now decreased to 1.2-1.3 x 10(6) CD34+ cells/kg. In whole blood transplantation of lymphoma patients (n = 45) receiving a BEAM-like regimen, low doses of CD34+ cells were sufficient for recovery (0.3-0.4 x 10(6)CD34+ cells/kg). In contrast to freeze-thawing of leucapheresis material, a high viability of CD34+ cells was preserved during storage for 3 days at 4 degrees C, leaving threshold doses for recovery unchanged. In conclusion, the Syto16 assay reveals the presence of many more non functional stem cells in frozen-thawed transplants than presumed thus far. This led to a factor 2.3-fold adjustment downward of viable CD34+ threshold doses for haematological recovery. PMID- 11859399 TI - Risk factors for post-stem cell transplant sinusitis. AB - An understanding of the factors that place the post-transplant patient at increased risk for sinusitis would help identify patients likely to develop disease and possibly allow for interventions that would decrease the incidence or severity of sinus disease. This retrospective study investigates the ability of screening paranasal sinus computed tomographic scans (CTs), clinical history, and potential risk factors for sinusitis, including history of tobacco use, history of allergies or asthma, IgG level, history of sinusitis, remission status and acute graft-versus-host disease (GVHD) to predict post-transplant sinusitis. Medical records and sinus CTs of 100 allogeneic bone marrow recipients were reviewed. There was no increased risk of developing sinusitis post SCT for patients with significant disease on screening CT, symptoms at time of transplant, a history of tobacco use, asthma or allergies, low IgG level, history of sinusitis or for patients at high risk of relapse. Patients with GVHD were 4.3 times more likely than patients without GVHD to develop sinusitis post transplant (95% CI: 1.7-11.0, P = 0.002). Acute GVHD places patients at greater risk of developing sinus infections. PMID- 11859400 TI - Valacyclovir prophylaxis for the prevention of Herpes simplex virus reactivation in recipients of progenitor cells transplantation. AB - HSV can cause oral lesions that exacerbate chemotherapy-related mucositis. Intravenous acyclovir is effective in preventing HSV reactivations, but expensive. Valacyclovir has good bioavailability and has not been studied for prophylaxis of HSV among PCT patients. We compared the efficacy and costs of valacyclovir in preventing HSV reactivation among HSV seropositive autologous progenitor cell transplantation (APCT) patients with historical controls in whom intravenous acyclovir or no HSV prophylaxis were used. Valacyclovir group: From October 1997 to April 1999 108 adult patients received valacyclovir 500 mg twice daily from day -3 of APCT until neutropenia recovery or day +30. Valacyclovir was switched to intravenous acyclovir in cases of oral intolerance (17 patients) or suspected HSV reactivation (five patients). Intravenous acyclovir group: From January 1996 to October 1997 43 patients received 5 mg/kg twice-daily intravenous acyclovir from day -3 until recovery from neutropenia. No prophylaxis group: 38 patients from January 1996 to October 1997 did not receive HSV prophylaxis. HSV reactivations were seen in 2.7%, 2% and 45% of patients in the valacyclovir, intravenous acyclovir, and no prophylaxis groups, respectively. Valacyclovir was well tolerated and was the least expensive strategy. Oral valacyclovir was as effective as intravenous acyclovir for the prophylaxis of HSV reactivation in APCT patients. PMID- 11859401 TI - Successful allogeneic bone marrow transplantation from an HBV-positive donor into an HBV-positive recipient using lamivudine. AB - A 21-year-old woman with severe aplastic anemia underwent allogeneic bone marrow transplantation from an HLA-identical sibling donor. The patient also had chronic hepatitis B and the donor was an HBV carrier. To decrease HBV and improve hepatic dysfunction before BMT, the patient had received lamivudine for 6 months. After marrow transfusion, administration of lamivudine was continued to inhibit replication of donor-derived HBV. The patient showed hematological engraftment on day 13 without any serious liver dysfunction. Eight months after BMT, she is now alive and well without chronic liver GVHD or reactivation of hepatitis B. HBV-DNA was not detected in the patient's serum. Administration of lamivudine to a BMT recipient with chronic hepatitis B may be a safe and promising way to prevent fatal liver dysfunction in the setting of allogeneic BMT, even in the event of BMT from an HBV-positive donor. PMID- 11859402 TI - High-dose chemotherapy and autologous CD34-positive blood stem cell transplantation for multiple myeloma in an HIV carrier. AB - The epidemiology and clinical outcome of multiple myeloma in human immunodeficiency virus (HIV)-positive patients is poorly documented. There are uncertainties concerning the optimal management of this rare disorder. We report on the use of myeloablative chemotherapy with autologous stem cell transplantation in an HIV-positive patient with multiple myeloma. PMID- 11859404 TI - Functional DNA array' in the fly: implication for neuronal degeneration. PMID- 11859403 TI - The two faces of p73. PMID- 11859405 TI - Lipids, cardiolipin and apoptosis: a greasy licence to kill. PMID- 11859406 TI - Role of p73 in malignancy: tumor suppressor or oncogene? AB - The recently identified p53 family member, p73, shows substantial structural and functional homology with p53. However, despite the established role of p53 as a proto-type tumor suppressor, a similar function of p73 in malignancy is questionable. Overexpression of p73 can activate typical p53-responsive genes, and activation of p73 has been implicated in apoptotic cell death induced by aberrant cell proliferation and some forms of DNA-damage. These data together with the localization of TP73 on chromosome 1p36, a region frequently deleted in a variety of human tumors, led to the hypothesis that p73 has tumor suppressor activity just like p53. However, unlike p53-/- mice, p73 knockout mice do not develop tumors. Extensive studies on primary tumor tissues have revealed overexpression of wild-type p73 in the absence of p73 mutations instead, suggesting that p73 may augment, rather than inhibit tumor development. In contrast to p53, differential splicing of the TP73 gene locus gives rise to a complex pattern of interacting p73 isoforms with antagonistic functions. In fact, induction of apoptosis by increased levels of p73 can be blocked by both p53 mutants and the N-terminally truncated p73 isoforms, which were recently shown to possess oncogenic potential. In the light of these new findings the contradictory role of p73 in malignancy will be discussed. PMID- 11859407 TI - Expression of DeltaNp73 is a molecular marker for adverse outcome in neuroblastoma patients. AB - The p73 gene is a p53 homologue which induces apoptosis and inhibits cell proliferation. Although p73 maps at 1p36.3 and is frequently deleted in neuroblastoma (NB), it does not act as a classic oncosuppressor gene. In developing sympathetic neurons of mice, p73 is predominantly expressed as a truncated anti-apoptotic isoform (DeltaNp73), which antagonizes both p53 and the full-length p73 protein (TAp73). This suggests that p73 may be part of a complex tumor-control mechanism. To determine the role of DeltaNp73 in NB we analyzed the pattern of expression of this gene in vivo and evaluated the prognostic significance of its expression. Our results indicate that DeltaNp73 expression is associated with reduced apoptosis in a NB tumor tissue. Expression of this variant in NB patients significantly correlates with age at diagnosis and VMA urinary excretion. Moreover it is strongly associated with reduced survival (HR=7.93; P<0.001) and progression-free survival (HR=5.3; P<0.001) and its role in predicting a poorer outcome is independent from age, primary tumor site, stage and MYCN amplification (OS: HR=5.24, P=0.012; PFS: HR=4.36, P=0.005). In conclusion our data seem to indicate that DeltaNp73 is a crucial gene in neuroblastoma pathogenesis. PMID- 11859409 TI - Identification of ter94, Drosophila VCP, as a modulator of polyglutamine-induced neurodegeneration. AB - We have successfully generated a Drosophila model of human polyglutamine (polyQ) diseases by the targeted expression of expanded-polyQ (ex-polyQ) in the Drosophila compound eye. The resulting eye degeneration is progressive and ex polyQ dosage- and ex-polyQ length-dependent. Furthermore, intergenerational changes in repeat length were observed in homozygotes, with concomitant changes in the levels of degeneration. Through genetic screening, using this fly model, we identified loss-of-function mutants of the ter94 gene that encodes the Drosophila homolog of VCP/CDC48, a member of the AAA+ class of the ATPase protein family, as dominant suppressors. The suppressive effects of the ter94 mutants on ex-polyQ-induced neurodegeneration correlated well with the degrees of loss-of function, but appeared not to result from the inhibition of ex-polyQ aggregate formation. In the ex-polyQ-expressing cells of the late pupa, an upregulation of ter94 expression was observed prior to cell death. Co-expression of ter94 with ex polyQ severely enhanced eye degeneration. Interestingly, when ter94 was overexpressed in the eye by increasing the transgene copies, severe eye degeneration was induced. Furthermore, genetical studies revealed that ter94 was not involved in grim-, reaper-, hid-, ced4-, or p53-induced cell death pathways. From these observations, we propose that VCP is a novel cell death effector molecule in ex-polyQ-induced neurodegeneration, where the amount of VCP is critical. Control of VCP expression may thus be a potential therapeutic target in ex-polyQ-induced neurodegeneration. PMID- 11859408 TI - Role of glutathione depletion and reactive oxygen species generation in apoptotic signaling in a human B lymphoma cell line. AB - The primary objective of this study was to determine the sequence of biochemical signaling events that occur after modulation of the cellular redox state in the B cell lymphoma line, PW, with emphasis on the role of mitochondrial signaling. L Buthionine sulphoximine (BSO), which inhibits gamma glutamyl cysteine synthetase (gammaGCS), was used to modulate the cellular redox status. The sequence and role of mitochondrial events and downstream apoptotic signals and mediators was studied. After BSO treatment, there was an early decline in cellular glutathione (GSH), followed by an increase in reactive oxygen species (ROS) production, which induced a variety of apoptotic signals (detectable at different time points) in the absence of any external apoptotic stimuli. The sequence of biochemical events accompanying apoptosis included a 95% decrease in total GSH and a partial (25%) preservation of mitochondrial GSH, without a significant increase in ROS production at 24h. Early activation and nuclear translocation of the nuclear factor kappa B subunit Rel A was observed at approximately 3h after BSO treatment. Cytochrome c release into the cytosol was also seen after 24h of BSO treatment. p53 protein expression was unchanged after redox modulation for up to 72 h, and p21waf1 independent loss of cellular proliferation was observed. Surprisingly, a truncated form of p53 was expressed in a time-dependent manner, beginning at 24h after BSO incubation. Irreversible commitment to apoptosis occurred between 48 and 72 h after BSO treatment when mitochondrial GSH was depleted, and there was an increase in ROS production. Procaspase 3 protein levels showed a time-dependent reduction following incubation with BSO, notably after 48 h, that corresponded with increasing ROS levels. At 96 h, caspase 3 cleavage products were detectable. The pan-caspase inhibitor zVADfmk, partially blocked the induction of apoptosis at 48 h, and was ineffective after 72 h. PW cells could be rescued from apoptosis by removing them from BSO after up to 48, but not 72 h incubation with BSO. Mitochondrial transmembrane potential (DeltaPsi(m)) remained intact in most of the cells during the 72 h observation period, indicating that DeltaPsi(m) dissipation is not an early signal for the induction of redox dependent apoptosis in PW cells. These data suggest that a decrease in GSH alone can act as a potent early activator of apoptotic signaling. Increased ROS production following mitochondrial GSH depletion, represents a crucial event, which irreversibly commits PW cells to apoptosis. PMID- 11859410 TI - Inflammatory cytokine regulation of TRAIL-mediated apoptosis in thyroid epithelial cells. AB - Death receptor-mediated apoptosis has been implicated in target organ destruction in chronic autoimmune thyroiditis. Depending on the circumstances, inflammatory cytokines such as IL-1, TNF and IFNgamma have been shown to contribute to either the induction, progression or inhibition of this disease. Here we demonstrate that the death ligand TRAIL can induce apoptosis in primary, normal, thyroid epithelial cells under physiologically relevant conditions, specifically, treatment with the combination of inflammatory cytokines IL-1beta and TNFalpha. In contrast, IFNgamma is capable of blocking TRAIL-induced apoptosis in these cells. This regulation of TRAIL-mediated apoptosis by inflammatory cytokines appears to be due to alterations of cell surface expression of TRAIL receptor DR5 and not DR4. We also show the in vivo presence of TRAIL and TRAIL receptors DR5 and DcR1 in both normal and inflamed thyroids. Our data suggests TRAIL-mediated apoptosis may contribute to target organ destruction in chronic autoimmune thyroiditis. PMID- 11859411 TI - Induction of apoptosis by chemotherapeutic drugs: the role of FADD in activation of caspase-8 and synergy with death receptor ligands in ovarian carcinoma cells. AB - Although ovarian tumours initially respond to chemotherapy, they gradually acquire drug resistance. The aims of this study were to identify how chemotherapeutic drugs with diverse cellular targets activate apoptotic pathways and to investigate the mechanism by which exposure to a combination of drugs plus death receptor ligands can increase tumour cell kill. The results show that drugs with distinct cellular targets differentially up-regulate TRAIL and TNF as well CD95L, but do not require interaction of these ligands with their receptor partners to induce cell death. Factors that were critical in drug-induced apoptosis were activation of caspases, with caspase-8 being activated by diverse drugs in a FADD-independent manner. Certain drugs also demonstrated some dependence on FADD in the induction of cell death. Caspase-9 was activated more selectively by chemotherapeutic agents. Combining ligation of death receptors with exposure to drugs increased tumour cell kill in both drug resistant cell lines and primary ovarian carcinoma cells, even though these cells were not sensitive to death receptor ligation alone. CD95L was more consistent at combining with drugs than TRAIL or TNF. Investigation of the mechanism by which a combination of drugs plus CD95 ligation can increase cell death showed that caspase-8 was activated in cells exposed to a combination of cisplatin and anti CD95, but not in cells exposed to either agent alone. PMID- 11859412 TI - A matrix-assisted laser desorption ionization post-source decay (MALDI-PSD) analysis of proteins released from isolated liver mitochondria treated with recombinant truncated Bid. AB - A crucial event in the process of apoptosis is caspase-dependent generation of truncated Bid (tBid), inducing release of cytochrome c. In an in vitro reconstitution system we combined purified recombinant tBid with isolated liver mitochondria and identified the released proteins using a proteomic matrix assisted laser desorption ionization post-source decay (MALDI-PSD) approach. In order to meet physiological conditions, the concentration of tBid was chosen such that it was unable to induce cytochrome c release in mitochondria derived from liver-specific Bcl-2-transgenic mice. Several mitochondrial proteins were identified to be released in a tBid-dependent way, among which cytochrome c, DIABLO/Smac, adenylate kinase 2, acyl-CoA-binding protein, endonuclease G, polypyrimidine tract-binding protein, a type-I RNA helicase, a WD-40 repeat containing protein and the serine protease Omi. Western blotting confirmed the absence of adenylate kinase 3, a matrix mitochondrial protein. These results demonstrate that a physiologically relevant concentration of tBid is sufficient to induce release of particular intermembrane mitochondrial proteins belonging to a broad molecular-mass range. PMID- 11859413 TI - Transglutaminase activity during senescence and programmed cell death in the corolla of tobacco (Nicotiana tabacum) flowers. AB - Corolla life span of undetached flowers of Nicotiana tabacum was divided into stages from the closed corolla (stage 1) through anthesis (stage 5) to death (stage 9). Senescence began around stage 6 in the proximal part, concomitantly with DNA laddering. Nuclear blebbing, DNA laddering, cell wall modification, decline in protein, water, pigment content and membrane integrity were observed during senescence and PCD. Transglutaminase activity was measured as mono- and bis-derivatives of putrescine (mono-PU; bis-PU) and bis-derivatives of spermidine (bis-SD). Bis-derivatives decreased with the progression of senescence, while mono-PU increased during early senescence; derivatives were present in different amounts in the proximal and distal parts of the corolla. In excised flowers, exogenous spermine delayed senescence and PCD, and caused an increase in free and acid-soluble conjugated PA levels. Bis-PU was the most abundant PA-derivative before DNA laddering stage; thereafter, bis-PU generally decreased and mono-PU became the most abundant derivative. PMID- 11859414 TI - Yeast two-hybrid screening using constitutive-active caspase-7 as bait in the identification of PA28gamma as an effector caspase substrate. AB - Caspase-3 and -7 represent executioner/effector caspases that directly cause apoptotic morphological changes by cleaving various death substrates. The substrates for caspases generally interact with active caspases, but not with inactive zymogens of caspase or procaspases. Here, to isolate proteins that interact with caspase-7, we established a yeast two-hybrid screening system using reversed-caspase-7, a constitutive active mutant of caspase-7 as a bait plasmid. Screening of an adult brain cDNA library led to isolation of proteasome activator 28 subunit, PA28gamma. In vitro translates of PA28gamma were cleaved by both recombinant caspase-3 and -7. Mutagenesis of potential cleavage site DGLD80 to EGLE80 completely abolished caspase-mediated cleavage. Moreover, endogenous PA28gamma was cleaved during not only Fas-induced apoptosis of HeLa cells, but also cisplatin-induced cell death of MCF7 cells, which are devoid of caspase-3. These findings indicate that PA28gamma is an endogenous substrate for caspase-3 and -7 and that yeast two-hybrid screening using reversed-caspase is a novel and useful approach to clone substrates for effector caspases. PMID- 11859415 TI - Potentiation of Fas-mediated apoptosis by an engineered glycosylphosphatidylinositol-linked Fas. AB - FasL and TRAIL are apoptotic ligands of the TNF-like cytokines family, acting via activation of the transmembrane death domain containing receptors Fas for FasL, and DR4 or DR5 for TRAIL. A glycosylphosphatidylinositol-linked TRAIL receptor called DcR1 behaves as a decoy receptor inhibiting TRAIL-mediated cell death in several cellular systems. We engineered and stably expressed a chimeric GPI linked Fas receptor (Fas-GPI) in T-lymphocyte cell lines constitutively expressing functional transmembrane Fas. Surprisingly, despite lacking the death domain region of functional Fas, Fas-GPI was able to significantly increase Fas mediated cell death triggered by membrane bound or soluble FasL, whereas engagement of Fas-GPI alone did not trigger apoptosis. This potentiating effect, but not transmembrane Fas activation, was selectively inhibited by protein kinase C activation with phorbol esters, demonstrating that Fas-GPI activated a specific synergistic signal transduction pathway. Fas-GPI and transmembrane Fas were localized in distinct membrane compartments, since Fas-GPI, but not transmembrane Fas, was found in the glycolipid-rich membrane microdomains. These results suggest that apoptosis induced by members of this ligand/receptors family may be differentially modulated through other and parallel signalling pathways. PMID- 11859417 TI - Role of methylation in the control of DeltaNp73 expression in neuroblastoma. PMID- 11859416 TI - Apaf1 reduced expression levels generate a mutant phenotype in adult brain and skeleton. PMID- 11859418 TI - Utilization of synthetic peptides containing nuclear localization signals for nonviral gene transfer systems. AB - The ability of nonviral gene delivery systems to overcome extracellular and intracellular barriers is a critical issue for future clinical applications. In recent years, several efforts were focused on the elucidation of the gene transfer mechanisms and on the development of multicomponent systems in order to improve both targeted gene delivery and transfection efficiency. The transport of the therapeutic DNA from the cytoplasm into the nucleus is an inefficient process and is considered as the major limiting step in nondividing cells. One of the strategies to improve nuclear uptake of DNA is taking advantage of the cellular nuclear import machinery. Synthetic peptides containing a nuclear localization signal (NLS) are bound to the DNA so that the resulting DNA-NLS complex can be recognized as a nuclear import substrate by specific intracellular receptor proteins. In this review, we critically summarize recent studies applying this approach with a particular focus on NLS-sequence specificity. Implications of the observed results are also discussed in regards to future developments of this technology. PMID- 11859419 TI - Tumor-specific transcriptional targeting of suicide gene therapy. AB - Transcriptional targeting of gene expression has been plagued by the weakness of tissue-specific promoters. Thus, to increase promoter strength while maintaining tissue specificity, we constructed a recombinant adenovirus containing a binary promoter system with a tumor-specific promoter (CEA; carcinoembryonic antigen) driving a transcription transactivator, which then activates a minimal promoter to express a suicide gene (HSV-tk; herpes simplex virus thymidine kinase). This ADV/binary-tk induced equal or greater cell killing in a CEA-specific manner in vitro compared with the CEA-independent killing of a vector with a constitutive viral promoter driving HSV-tk (ADV/RSV-tk). To monitor adenovirus-mediated HSV-tk gene expression in vivo, we employed noninvasive nuclear imaging using a radioiodinated nucleoside analog ([((1)31)I]-FIAU) serving as a substrate for HSV tk. [((1)31)I]-FIAU-derived radioactivity accumulated after intratumoral injection of ADV/binary-tk only in the area of CEA-positive tumors with significantly less spread to the adjacent liver tissue than after administration of the universally expressed ADV/RSV-tk. Both viruses exhibited similar antitumor efficacy upon injection of liver metastases. Importantly, in vivo dose escalation studies demonstrated significantly reduced toxicity after intravenous administration of ADV/binary-tk versus ADV/RSV-tk. In summary, the increased therapeutic index of this novel, amplified CEA-driven suicide gene therapy vector is a proof of principle for the powerful enhancement of a weak tissue-specific promoter for effective tumor restricted gene expression. PMID- 11859420 TI - Redirecting adenovirus to pulmonary endothelium by cationic liposomes. AB - Somatic gene transfer to the pulmonary endothelium may be a useful strategy for modifying the phenotype of endothelium and/or vascular smooth muscle in disorders such as primary pulmonary hypertension, ARDS or pulmonary metastatic disease. Adenoviral (Ad) vectors, although highly efficient in liver gene transfer, have proven to be limited for pulmonary gene transfer with respect to efficiency, in part because of difficulty in assuring significant residence time in the lung and/or paucity of receptors for adenovirus on the endothelium. A recent study has shown that the use of a bispecific antibody to endothelial cells and Ad vectors efficiently redirects Ad vectors to pulmonary endothelium and improves gene expression in the lung. In this study, we report that pulmonary gene transfer by Ad vectors can also be improved significantly via the use of cationic liposomes. Preinjection of cationic liposomes followed by adenovirus led to a significant increase in the level of gene expression in the lung. The improvement in pulmonary gene transfer was associated with a decrease in the level of gene expression in the liver. Gene expression in the lung lasted for up to 2 weeks. This protocol, together with genetic modification of adenovirus, may prove to be useful for pulmonary gene transfer for the treatment of pulmonary diseases. This method may also be extended to pulmonary gene transfer using other types of viral vectors via vascular route. PMID- 11859421 TI - Viral gene delivery of superoxide dismutase attenuates experimental cholestasis induced liver fibrosis in the rat. AB - Hydrophobic bile acids lead to generation of oxygen free radicals in mitochondria. Accordingly, this study investigated if gene delivery of superoxide dismutase (SOD) would reduce hepatic injury caused by experimental cholestasis. Rats were given adenovirus (Ad; 3 x 10(9) p.f.u., i.v.) carrying the bacterial control gene lacZ, mitochondrial Mn-SOD or cytosolic Cu/Zn-SOD genes 3 days before bile duct ligation. Both Mn- and Cu/Zn-SOD activity was increased in the liver about four-fold 3 days after viral infection. Serum alanine transaminase increased to about 710 U/l after bile duct ligation, which was blunted by about 70% in rats receiving Ad-Mn-SOD, but by only 30% in rats receiving Ad-Cu/Zn-SOD. Bile duct ligation caused focal necrosis, apoptosis and fibrosis in the liver and increased collagen alpha1 mRNA about 20-fold. These effects were reduced significantly by Ad-Mn-SOD, but not by Ad-Cu/Zn-SOD. In addition, bile duct ligation increased 4-hydroxynonenal, a product of lipid peroxidation, activated NF-kappaB and increased synthesis of TNF(alpha) and TGF-beta. These effects were also blunted significantly by Ad-Mn-SOD, but not by Ad-Cu/Zn-SOD. Taken together, it is concluded that cholestasis causes liver injury by mechanisms involving mitochondrial oxidative stress. Gene delivery of mitochondrial Mn-SOD blocks formation of oxygen radicals and production of toxic cytokines thereby minimizing liver injury caused by cholestasis. PMID- 11859422 TI - Adenovirus-mediated gene transfer of urokinase plasminogen inhibitor inhibits angiogenesis in experimental arthritis. AB - Plasmin is essential for metalloproteases activation, endothelial cell migration and degradation of the extracellular matrix. The process is common to neoangiogenesis pannus formation and cartilage degradation within arthritic joints. Since 80% of synovial cells express urokinase plasminogen activator receptor (uPAR), we investigated the inhibition of plasmin activation in a collagen-induced arthritis (CIA) mice model, by expressing a uPA/uPAR antagonist molecule (ATF) fused to human serum albumin (HSA) to extend its serum half-life. Overexpression was obtained with an adenoviral vector expressing the chimeric murine ATF-HSA. We showed that the genetic coupling did not significantly reduce the ability of the ATF moiety to interact with its receptor uPAR. The chimeric protein was detectable in the sera of injected mice 7 days following Ad-mATF-HSA injection, then decreased in parallel with the anti-HSA titer increase. Systemic Ad-mATF-HSA injection performed on day 25 following CIA induction decreased the incidence of arthritis and the severity of the disease. Moreover, synovial angiogenesis in arthritic paws was decreased after Ad-mATF-HSA gene transfer, as assessed by smooth muscle actin immunostaining. The preventive effect observed on arthritis was related to the decrease in angiogenesis, rather than inhibition of extracellular matrix degradation. PMID- 11859423 TI - Effect of combined cytostatic cyclosporin A and cytolytic suicide gene therapy on the prevention of experimental graft-versus-host disease. AB - The immunosuppressive drug cyclosporin A (CsA) represents the standard preventive treatment of graft-versus-host disease (GVHD), the main complication of allogeneic hematopoietic stem cell transplantation (HSCT). However, its efficacy is only partial and many patients develop lethal GVHD despite CsA. A strategy of genetic immunosuppression based on conditional elimination of donor T cells expressing the Herpes simplex type 1 thymidine kinase (TK) suicide gene was recently developed. In this system, ganciclovir (GCV) selectively kills dividing but not quiescent TK T cells. Since CsA is known to have a cytostatic effect on T cells, it could negatively interfere with the division-dependent TK gene therapy. We thus tested whether administration of CsA would antagonize elimination of alloreactive donor TK T cells mediated by GCV in a murine model of GVHD. In vivo experiments revealed that, contrary to GCV, CsA only transiently controlled alloactivation-induced T cell proliferation, and likewise could not prevent lethal GVHD. When T cells resumed proliferation under CsA, they were however still sensitive to GCV. Survival, as well as immune reconstitution, was excellent in mice treated with GCV alone or in combination with CsA. These observations should help to design improved suicide gene therapy trials in the field of allogeneic HSCT. PMID- 11859424 TI - Genetic immunization with a melanocytic self-antigen linked to foreign helper sequences breaks tolerance and induces autoimmunity and tumor immunity. AB - Mechanisms maintaining peripheral tolerance to self-antigens present a major obstacle for the development of antigen-specific melanoma vaccines, presumably because self-antigens are not able to stimulate a CD4 T-helper response. Using the melanosomal enzyme tyrosinase-related protein 2 (TRP2) expressed by melanocytes and most melanoma cells as a model self-antigen, we investigated whether linkage with a foreign immunogenic protein providing strong CD4 helper sequences would be able to circumvent tolerance and enhance the induction of antigen-specific tumor immunity. We found that genetic immunization of mice with cDNA encoding a fusion protein between enhanced green fluorescent protein (EGFP) from jellyfish and autologous murine TRP2 (EGFP.mTRP2) resulted in the stimulation of TRP2-reactive T cells in vivo. Importantly, immunization with EGFP.mTRP2 effectively protected mice against metastatic growth of B16 melanoma in the lungs and was associated with fur depigmentation as a sign of autoimmune mediated destruction of melanocytes. Our results show that tumor vaccines consisting of self-antigens linked to immunogenic helper sequences can be successfully applied to the immunotherapy of melanoma and provide a scientific basis for the translation of this strategy in future clinical investigations. PMID- 11859426 TI - Adenoviral transfer of a single donor-specific MHC class I gene to recipient bone marrow cells can induce specific immunological unresponsiveness in vivo. AB - We investigated the delivery of a donor-specific MHC class I gene, H-2K(b), using a newly constructed replication-defective recombinant adenovirus (AdSV40K(b)) to recipient tissue before transplantation as a means of inducing donor-specific immunological unresponsiveness. AdSV40K(b) was able to transduce both a fibroblast cell line and freshly isolated bone marrow cells (BMCs) resulting in cell surface expression of H2-K(b) protein. Intravenous infusion of AdSV40K(b) transduced syngeneic CBA/Ca (H-2(k)) BMCs into CBA recipient mice treated with an anti-CD4 monoclonal antibody 27 days before transplantation of a fully MHC mismatched, C57BL/10 (H-2K(b+)), cardiac allograft resulted in significant long term graft survival when compared with mice receiving the same dose of syngeneic BMCs transduced with a control adenovirus, AdRSVbetagal. Despite the induction of H-2K(b)-specific hyporesponsiveness following pretreatment with AdSV40K(b) transduced CBA BMCs, persistence of H-2K(b) mRNA in central or peripheral tissues could not be demonstrated by RT-PCR. This result was in contrast to the observed persistence of K(b) mRNA both in the periphery and thymus following the infusion of transgenic CBK (H-2(k) + K(b)) BMCs. We conclude that ex vivo adenoviral gene transfer of a single donor MHC class I gene to recipient BMCs in combination with transient depletion of CD4(+) cells is sufficient to induce long-term graft survival of a fully allogeneic cardiac graft. In addition, detectable microchimerism is not a prerequisite for graft survival. PMID- 11859425 TI - HSV-mediated delivery of virally derived anti-apoptotic genes protects the rat hippocampus from damage following excitotoxicity, but not metabolic disruption. AB - Studies utilizing gene delivery to the nervous system indicate that various strategies are protective following acute neurological insults such as seizure and stroke. We have found that inhibitors of apoptosis are protective against excitotoxicity and heat stress but not energetic impairment in vitro. Here we studied the neuroprotective efficacy in vivo of these mediators: viral genes (crmA, p35, gamma34.5 KsBcl-2) that have evolved to suppress suicidal host responses to infection, by inhibiting apoptosis. We investigated these effects by utilizing modified herpes vectors to deliver the anti-apoptotic agents intracerebrally and examined them in the face of excitotoxic and metabolic insults. We found that p35 and gamma34.5 reduced by 45% a hippocampal CA3 lesion caused by kainic acid, while crmA and KsBcl-2 did not. None of the inhibitors protected the dentate gyrus of the hippocampus following 3-acetylpyridine, a hypoglycemia model, but we found crmA to worsen the damage. These data are similar to our results in neuronal cultures where the inhibitors protected against the excitotoxin domoic acid, but not against the metabolic poison, cyanide. Together, the results suggest that inhibitors of various apoptotic elements are capable of protecting under acute insult conditions both in vitro and in vivo, suggesting possible future therapeutic applications. PMID- 11859427 TI - Potential limitations of transcription terminators used as transgene insulators in adenoviral vectors. AB - The presence of adenoviral cis-elements interfering with the activity of tissue specific promoters has seriously impaired the use of transcriptional targeting adenoviruses for gene therapy purposes. As an approach to overcome this limitation, transcription terminators were previously employed in cultured cells to insulate a transgene promoter from viral activation. To extend these studies in vivo, we have injected into heart and skeletal muscle, adenoviruses containing the human growth hormone terminator and the cardiac-specific alpha-myosin heavy chain promoter (alphaMyHC) driving the chloramphenicol acetyltransferase (CAT) reporter gene. Promoterless CAT constructs were also tested to study interfering viral transcription and terminator activity. Here we demonstrate that the presence of a terminator can produce undesirable effects on the activity of heterologous promoters. Our analysis shows that in particular conditions, a terminator can reduce the tissue specificity of the transgene promoter. By RNAse protection assay performed on cardiac myocytes, we also show that adenoviral elements can direct high levels of autonomous transcription within the E1A enhancer region. This finding supports the model that passive readthrough of the transgene promoter is responsible for loss of selective expression. PMID- 11859428 TI - Comparison of extremely low frequency (ELF) magnetic field personal exposure monitors. AB - The MultiWave System III (MW III), a recently developed personal monitor for extremely low frequency (ELF) magnetic fields, was compared with the standard EMDEX Lite (Electric and Magnetic Field Digital Exposure System), the type of monitor widely used in epidemiology and other exposure assessments. The MW III captures three-axis magnetic field waveforms for the calculation of many exposure metrics, while the EMDEX monitors measure only the root-mean-squared (RMS) vector magnitude (or resultant). Thirty-eight partial period personal samples were monitored in six different job classifications. The sampling time for each personal sample ranged from 90 to 133 min, with a mean sample time of 110 min. The EMDEX Lite and MW III were evaluated by comparing the maximum and partial period time-weighted average (TWA) of the ELF magnitude. TWA exposures measured for the 38 partial period samples by the EMDEX Lite ranged from 1.2 to 65.3 mG, with a mean of 18.1 mG, while corresponding values for the MW III ranged from 1.1 to 65.8 mG, with a mean of 17.7 mG. The maximum magnetic field exposures measured for the 38 partial period personal samples by the EMDEX Lite ranged from 27.0 to 420.2 mG, with a mean of 216.3 mG, while corresponding values for the MW III ranged from 40.2 to 1311.8 mG, with a mean of 368.4 mG. The maximum and TWA ELF magnetic field exposures measured by the EMDEX Lite and MW III were compared using a two-tailed, paired t-test. Analyses indicate that there was no significant difference in the TWA magnetic field magnitude measured by the EMDEX Lite and MW III. On the other hand, the EMDEX Lite reported significantly lower (P=0.002) maximum magnetic field measurements compared to the MW III. From a detailed analysis of the time traces, the EMDEX Lite appears to measure the ELF magnitude inaccurately when the field changes rapidly over a 4-s sampling interval. The results of this comparison suggest that the standard EMDEX Lite and MW III provide similar measure of the TWA magnetic field in a variety of occupational settings and ELF magnetic field magnitudes. However, the EMDEX Lite underestimates maximum exposures when compared to the MW III. PMID- 11859429 TI - Results of a multisite study of U.S. residential magnetic fields. AB - This paper describes the study design, measurement protocols, and results of a project examining residential magnetic-field exposures at eight sites across the contiguous United States. The goal of the project was to investigate surrogates that have been used in epidemiologic studies to characterize residential magnetic field exposure. These surrogates include: personal-exposure (PE), fixed-location long-term (LT), and outside and inside point-in-time (PIT) magnetic-field measurements; net-service (or ground current) measurements; and the "wire-code category" of the residence. (The latter is a surrogate for magnetic-field exposure based on the nature and proximity of electric power lines outside the house.) Measurements were conducted on four visits to each of eight sites between January 1994 and June 1997 for a study population of 218 single-unit detached dwellings. Information on the residence, residents, and neighborhood was collected. A simple random sample of 392 single-unit detached dwellings at the sites was used to create a weighted sample of houses representative of the population of single-unit residences. The correlations among the various types of 60-Hz magnetic-field measures were relatively strong (Pearson r>0.74, Spearman rho>0.78). Variability of PE and LT measurements, as measured by the standard deviations during a visit, was independent of wire-code category. Visit means for PE, LT, and outside and inside PIT were well correlated over periods between visits of from 1.5 to 20 months (r>0.62, rho>0.76). These results support the use of survey measurements (less demanding than personal monitoring) to represent exposure that occurred up to 20 months in the past. The principal component of the total variance in PE measurements was the between-house variance; between visit and between-site variances were generally less important. This supports the sampling of many houses with relatively few visits in residential exposure characterization studies. There was a trend for presumably higher wire-code categories to be associated with higher field summary measures for all summary measures related to magnetic-field magnitude, including PE and LT resultant, and inside and outside resultant (60 Hz) and harmonics. However, because of the overlap in field levels between categories, wire code was not a good predictor of magnetic-field levels, accounting for less than 21% of the variance in magnetic field measurements. PMID- 11859430 TI - Assessment of organophosphorous pesticide exposures in the diets of preschool children in Washington State. AB - Twenty-four hour duplicate diet sampling was employed to investigate dietary pesticide exposures of children aged 2 to 5 years. Duplicate diets were collected from seven children living in the Seattle metropolitan area and six children living in Chelan and Douglas counties in Central Washington. Diet samples were collected from each child in the summer and again in the fall, and total daily diets were divided into four food categories: fresh fruits and vegetables, beverages, processed foods, and dairy products. A total of 88 individual food category samples were collected and analyzed for 15 organophosphorous (OP) pesticides. Three of the 13 children had no detectable OP pesticides in either of their diet samples, and 14 of the 26 duplicate diets did not contain detectable levels of OP pesticides. Sixteen individual food category samples contained detectable levels of at least one OP pesticide and two of these samples contained detectable levels of two OP pesticides. Of the 15 targeted pesticides, 6 were detected: azinphosmethyl, chlorpyrifos, malathion, methidathion, methyl parathion, and phosmet. Azinphosmethyl was detected most frequently (10% of all samples), particularly in samples containing apples or apple juice. The fresh fruits and vegetable category had the most frequent pesticide determinations, followed by beverages. OP pesticides were not present at detectable levels in any of the dairy samples. Malathion was the only OP pesticide detected in processed food samples, appearing in 4 of the 26 samples (15%). No detections were above the legal tolerances for residues on produce, however the acute population adjusted reference dose (aPAD) for chlorpyrifos exposure of 1.7 microg/kg/day was exceeded by one subject during one sampling event. This subject's cumulative daily dose of chlorpyrifos equivalents was estimated to be 2.5 microg/kg/day. PMID- 11859431 TI - National Human Exposure Assessment Survey: analysis of exposure pathways and routes for arsenic and lead in EPA Region 5. AB - The National Human Exposure Assessment Survey (NHEXAS) Phase I field study conducted in EPA Region 5 (Great Lakes Area) provides extensive exposure data on a representative sample of approximately 250 residents of the region. Associated environmental media and biomarker (blood, urine) concentration data were also obtained for the study participants to aid in understanding of the relationships of exposures to both contaminant pathways and doses. Besides fulfilling the primary NHEXAS objectives, the NHEXAS data provided an opportunity to explore secondary usages, such as examining pathway to route of exposure relationships. A generic type of structural equation model was used to define the anticipated relationships among the various data types for both arsenic (As) and lead (Pb). Since, by design, only a few participants provided data for all sample types, implementing this model required that some media concentrations (outdoor air and soil) be imputed for subjects with missing information by using measurements collected in the same geographic area and time period. The model, and associated pairwise correlations, generally revealed significant but weak associations among the concentrations, exposures, and doses; the strongest associations occurred for the various air measurements (indoor versus outdoor and personal). The generally weak associations were thought to be partly due to the absence of complete coverage of nonresidential environmental media and to nonsynchronization of relevant measurement times and integration periods of collection across the various sample types. In general, relationships between the NHEXAS questionnaire data and the various concentration, exposure, and body-burden measures were also weak. The model results and the modeling exercise suggest several ways for optimizing the design of future exposure assessment studies that are aimed at supporting structural modeling activities. PMID- 11859432 TI - Pharmacokinetics of 2,3,7,8-tetrachlorodibenzo-p-dioxin in Seveso adults and veterans of operation Ranch Hand. AB - A combined analysis of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) elimination in Seveso adults and Ranch Hand veterans found a period of fast elimination within the first 0.27 years after exposure in Seveso, followed by a period of slower elimination between 3 and 16.35 years from exposure. The mean TCDD elimination rate within the first 0.27 years after exposure among six adult males in the Seveso cohort was 2.0646 year(-1) (half-life=0.34 years). The mean rate from 3 to 16.35 years was 0.1011 year(-1) (half-life=6.9 years). The mean Ranch Hand elimination rate, 00924 year(-1) (half-life=6.9 years), measured between 9 and 33 years after exposure, was significantly less than the Seveso mean in the first 0.27 years after exposure, but not significantly different from the Seveso mean between 3 and 16.35 years after exposure. The fast elimination within the first 0.27 years followed by a slower rate after 3 years is consistent with the expected pattern in a two-compartment open model, with a distribution phase of rapid elimination followed by a slower elimination phase. PMID- 11859433 TI - Exposure methodology and findings for dietary nitrate exposures in children of Transylvania, Romania. AB - AIMS: The primary focus of this exposure assessment work involved developing an exposure model and determining a numerical point estimate of the amount of biologically relevant nitrate/nitrite exposure that occurred for each child in the study. This assessment was done in support of two epidemiological studies. The first study was an epidemiological cohort study (three cohorts based on nitrate/nitrite exposure) that explored the relationship between high nitrate/nitrite exposure and neuropsychological development. The second study was a nested case/control study (cases of methemoglobinemia versus disease-free controls) that sought to explore the relationship between MHG and various risk factors for the disease. METHODS: This work uses both dietary survey and environmental sampling and modeling in order to develop two point estimates of nitrate exposure in milligrams per kilogram per day of nitrite (the biologically active form of the hemoglobin-oxidizing agent) for the first 6 months of the child's life (2-months-of-age and 6-months-of-age point estimates). Methodologies included proxy interviews of primary caregivers, review of existing medical and environmental sampling and analysis. RESULTS: Exposure to nitrate--nitrogen (with final calculations converted to the biologically active form of the toxin, nitrite) was categorized as high, medium, and low as determined from the distribution of the data derived from final exposure calculations at both the 2 months-of-age point estimate and at the 6-months-of-age point estimate. These tertiles correspond to greater-than-or-equal1.5 mg/kg/day nitrite-nitrogen for high-exposure individuals, <1.5-> or = 0.1 mg/kg/day for medium-exposure individuals, and <0.1 mg/kg/day for low-exposure individuals. Analyses illustrate that over 90% of the nitrate exposure occurred through the consumption of liquids (water) at the 2-months-of-age point estimate while at the 6-months-of-age point estimate, a 10-fold change in the amount of solid consumables occurred. CONCLUSIONS: Final exposure calculations were well differentiated into three tertiles based on a point estimate of average daily intake of nitrite in milligrams per kilogram body weight per day at roughly 2 and 6 months of age. These categories of exposure, based on the exposure model point estimate, correspond well with the exposure estimates as estimated only on the basis of cohort status and their corresponding nitrate/nitrite well water levels. Comparisons of these two sets of data illustrate that following the MHG incident, Cohort II shifted places with Cohort I to become the high-exposure cohort. Further, the predictive ability of the exposure assessment in regard to the outcome of MHG was estimated using a Likelihood Ratio and Pearson's Crosstab analysis. This was performed on the 2-month-of-age point estimate. Likelihood Ratio and Pearson's chi-square were 39.40 and 33.74, respectively, with a probability of achieving these fits by chance alone of <0.0001. This indicates clearly that the children who experienced MHG were also the children at the 2 month-of-age point estimate who had received the highest exposure to nitrate/nitrite through their diet. PMID- 11859434 TI - Using GIS and historical records to reconstruct residential exposure to large scale pesticide application. AB - Investigation of pesticide impacts on human health depends on good measures of exposure. Historical exposure data are needed to study health outcomes, such as cancer, that involve long latency periods, and other outcomes that are a function of the timing of exposure. Environmental or biological samples collected at the time of epidemiologic study may not represent historical exposure levels. To study the relationship between residential exposure to pesticides and breast cancer on Cape Cod, Massachusetts, historical records of pesticide use were integrated into a geographic information system (GIS) to estimate exposures from large-scale pesticide applications between 1948 and 1995. Information on pesticide use for gypsy moth and other tree/vegetative pest control, cranberry bog cultivation, other agriculture, mosquito control, recreational turf management, and rights-of-way maintenance is included in the database. Residents living within or near pesticide use areas may be exposed through inhalation due to drift and volatilization and through dermal contact and ingestion at the time of application or in later years from pesticides that deposit on soil, accumulate in crops, or migrate to groundwater. Procedures were developed to use the GIS to estimate the relative intensity of past exposures at each study subject's Cape Cod addresses over the past 40 years, taking into account local meteorological data, distance and direction from a residence to a pesticide use source area, size of the source area, application by ground-based or aerial methods, and persistent or nonpersistent character of the pesticide applied. The resulting individual-level estimates of relative exposure intensity can be used in conjunction with interview data to obtain more complete exposure assessment in an epidemiologic study. While the database can improve environmental epidemiological studies involving pesticides, it simultaneously illustrates important data gaps that cannot be filled. Studies such as this one have the potential to identify preventable causes of disease and guide public policies. PMID- 11859435 TI - The effect of relevant genotypes on PAH exposure-related biomarkers. AB - Polycyclic aromatic hydrocarbons (PAHs) in coke oven emissions cause a cancer risk to humans. In a comprehensive biomonitoring study among Estonian coke oven workers, we looked at the effect of genetic polymorphisms in metabolic enzymes on urinary mutagenicity, 1-hydroxypyrene (1-OHP) concentration in urine, and aromatic DNA adducts in white blood cells (WBCs). Coke oven workers were sampled twice (samplings I and II), and controls only once at the time of sampling I. Urinary mutagenicity was measured using the Ames test. CYP1A1, microsomal epoxide hydrolase (mEH), and glutathione S-transferase (GST) genotypes were analyzed by polymerase chain reaction (PCR). Urinary mutagenicity did not differ between exposed and controls, but those coke oven workers who were smokers had significantly higher (P=0.0002) mutagenic activity in urine than nonsmokers. Urinary mutagenicity was moderately correlated to levels of 1-OHP and aromatic DNA adducts, the P values ranging from 0.0005 to 0.002. Carriers of a variant allele in exon 4 of mEH (Arg139) had elevated urinary mutagenicity (sampling I). In addition, urine mutagenicity of persons with predicted high mEH activity was significantly higher. Smoking habit did not explain the differences observed in urinary mutagenicity between mEH phenotype or genotype subgroups. Variation in exon 3 of mEH (His113) was related to a significantly (P=0.01) higher 1-OHP concentration in exposed workers (sampling II). Workers from sampling I who had an Arg139 variation in mEH had lower levels of adducts in lymphocytes (P=0.01) than others, while airborne benzo[a]pyrene (B[a]P) and His113 variation affected interactively on adduct levels. Our study shows that a comprehensive assessment of exposure is essential for elucidation of PAH exposure at a workplace. Even at high exposures metabolic polymorphisms seem to have some effect on biomarker levels, and should be assessed in biomonitoring studies. PMID- 11859436 TI - Experimental modelling of human spinal cord injury: a model that crosses the species barrier and mimics the spectrum of human cytopathology. AB - STUDY DESIGN: Literature review and presentation of an experimental model of human spinal cord injury, (SCI). OBJECTIVES: Experimental designs seek to mimic and model the physical processes by which human SCI occurs and replicate the variety of chronic pathologies that characterize its long term effects. The variations in biological processes that are present between species have contributed to recent difficulties in generalizing experimental findings to the human condition. In this review, one finds: (1) a discourse on the pathological nature of the chronic human lesion, (2) a consideration of how the physical properties of soft tissue injury result in acute and chronic changes in the spinal substance, (3) a description of a device (ESCID) that is able to replicate and dynamically monitor physical indices of SCI as they take place in experimental models, and (4) a summary of how use of this device in different species has allowed the biomechanical descriptors of such injuries to be easily compared even in murine models. SETTING: Ohio State University, Ohio, USA. RESULTS: Careful attention to the details of injury device design has finally allowed a direct comparison of contusion-type injury models in the rat and mouse. Biomechanical outcomes with predictive capabilities have evolved that allow the investigator to create the range of pathologies seen in the human lesion even in these small vertebrates. The predictive cytopathology and our ability to manipulate the mouse genome will allow the testing of specific hypotheses related to cause and effect in experimental spinal cord injuries. Since the biomechanics, pathology, and chronic outcomes appear to be similar to those seen in the human, these animal models should facilitate rapid progress in the design of human therapeutics. CONCLUSIONS: Biomechanics of certain elements of experimental spinal injury are surprisingly accurate descriptors of acute and chronic pathologies in the spinal cord. This tenet applies across species and has often allowed more accurate design of clinical trials in the past few decades. As molecular approaches to this problem evolve, the use of species with known genomes appear warranted. Models that take advantage of these approaches are likely to produce innovations that quicken the pace of human trial strategies. PMID- 11859437 TI - Improved glucose tolerance and insulin sensitivity after electrical stimulation assisted cycling in people with spinal cord injury. AB - DESIGN: Longitudinal training. OBJECTIVES: The purpose was to determine the effect of electrical stimulation (ES)-assisted cycling (30 min/day, 3 days/week for 8 weeks) on glucose tolerance and insulin sensitivity in people with spinal cord injury (SCI). SETTING: The Steadward Centre, Alberta, Canada. METHODS: Seven participants with motor complete SCI (five males and two females aged 30 to 53 years, injured 3-40 years, C5-T10) underwent 2-h oral glucose tolerance tests (OGTT, n=7) and hyperglycaemic clamp tests (n=3) before and after 8 weeks of training with ES-assisted cycling. RESULTS: Results indicated that subjects' glucose level were significantly lower at 2 h OGTT following 8 weeks of training (122.4+/-10 vs 139.9+/-16, P=0.014). Two-hour hyperglycaemic clamps tests showed improvement in all three people for glucose utilisation and in two of three people for insulin sensitivity. CONCLUSIONS: These results suggested that exercise with ES-assisted cycling is beneficial for the prevention and treatment of Type 2 diabetes mellitus in people with SCI. SPONSORSHIP: Supported by Alberta Paraplegic Foundation, Therapeutic Alliance. PMID- 11859438 TI - Pain classification following spinal cord injury: the utility of verbal descriptors. AB - OBJECTIVES: To determine the predictive utility of verbal descriptors to distinguish between pain types following spinal cord injury (SCI). DESIGN: Cross sectional. SETTING: USA. METHODS: Participants (n=29) completed the Short Form - McGill Pain Questionnaire (SF-MPQ) for each pain site reported. A total of 64 pain sites were reported with 80% of the sample reporting multiple pain sites. Each pain site was categorized using three different SCI pain classification schemes. The predictive utility of verbal descriptors to distinguish between pain types was examined statistically using (1) each word separately, (2) a combination of words (ie, the SF-MPQ total subscales, the number of words chosen on each scale), and (3) discriminant function analysis. RESULTS: There was a substantial overlap in the use of verbal descriptors across pain types. Few differences across pain types were found for endorsement of individual words, and differences across pain types were not found for any of the word combination scores. The majority of the verbal descriptors did not enter the step-wise discriminant functions for each SCI pain classification scheme, however, 'tingling' and 'aching' showed modest predictive utility for neuropathic and musculoskeletal pain, respectively. Correct classification was in the low range (ie, 39% to 82%, average=60%, with a 33% chance level). All three pain classification schemes showed the same general pattern of results. CONCLUSION: In general, verbal descriptors alone offered marginal utility with regard to identifying specific pain types following SCI. Future directions alone and implications are discussed. PMID- 11859439 TI - International differences in ageing and spinal cord injury. AB - DESIGN: The present study is part of a programme of longitudinal research on ageing and spinal cord injury involving three populations - American, British and Canadian. The design was multivariate. OBJECTIVE: To identify international differences in outcomes associated with ageing and spinal cord injury. SETTING: A sample of 352 participants was assembled from five large, well-established databases. The Canadian sample was derived from the member database of the Canadian Paraplegic Association (Ontario and Manitoba divisions). The British sample was recruited from Southport Hospital's Northwest Regional Spinal Injuries Centre and Stoke-Mandeville Hospital's National Spinal Injuries Centre. The American sample has been recruited through Craig Hospital in Denver, Colorado. METHODS: The sample included individuals who had incurred a spinal cord injury at least 20 years previously; were admitted to rehabilitation within 1 year of injury; were between age 15 and 55 at the time of injury. Data were collected using a combination of self-completed questionnaires and interviews. Data included medical information, general health, hospitalisations, and changes in bladder and bowel management, equipment, pain, spasticity, the need for assistance, and other health issues. RESULTS: Clear international differences existed between the three samples in the three different countries. After controlling for sampling differences (ie, differences in age, level of lesion, duration of disability, etc.), the following differences were seen: (1) American participants had a better psychological profile and fewer health and disability related problems; (2) British participants had less joint pain and less likelihood of perceiving they were ageing more quickly; (3) Canadians had more health and disability-related complications (particularly bowel, pain and fatigue problems). CONCLUSION: These differences are discussed in terms of socio political, health care system and cultural factors that might be used to explain them, and to generate hypotheses for future research. PMID- 11859440 TI - Re-aggravation of myelopathy due to intramedullary lesion with spinal cord enlargement after posterior decompression for cervical spondylotic myelopathy: serial magnetic resonance evaluation. AB - STUDY DESIGN: A case report. OBJECTIVES: To report a case of swelling of the spinal cord and an intramedullary lesion occurring after expansive laminoplasty for cervical spondylotic myelopathy. SETTING: A university hospital in Japan. METHODS: Clinical evaluation, radiography, MR imaging. RESULTS: A 65-year-old man with a cervical spondylotic myelopathy in whom symptoms were improved immediately after expansive laminoplasty, but became aggravated 2 weeks later. Magnetic resonance images demonstrated swelling of the spinal cord and an intramedullary lesion that extended from the medulla oblongata to C7. Nine months after surgery, the lesion was reduced to C2-6, but neurological deterioration had not improved. Six years after surgery, the patient remains confined to bed. CONCLUSION: Patients with such disease conditions are rare, and it is difficult to predict postoperative swelling of the spinal cord before surgery. Spine surgeons should be aware of such rare disease conditions involving the spinal cord. PMID- 11859441 TI - Autonomic dysreflexia associated with transient aphasia. AB - STUDY DESIGN: Case report of autonomic dysreflexia presenting with transient aphasia in a subject with C4 tetraplegia. OBJECTIVES: To report a rare case of autonomic dysreflexia. SETTING: Rehabilitation Service, The Ohio State University, USA. CASE REPORT: A 21-year-old man with a C4 spinal cord injury (ASIA B) developed aphasia associated with autonomic dysreflexia. He was treated with an adrenergic blocking agent. CONCLUSION: Autonomic dysreflexia manifested by a transient aphasia and seizures is uncommon. PMID- 11859442 TI - Cytomegalovirus-associated transverse myelitis in a non-immunocompromised patient. AB - DESIGN: Single case report. OBJECTIVE: To report a rare case of cytomegalovirus (CMV)-associated transverse myelitis (TM) in the immunocompetent host. SETTINGS: Collaboration between a Neurology and Radiology University Department in Greece and a Molecular Virology Department in Cyprus. PATIENT: A 16-year-old male student developed an acute febrile illness followed shortly by TM, that resulted in paraplegia over 24 h. Rapid clinical improvement was followed by complete recovery in 2 months. Extensive laboratory work-up excluded other possible causes of TM and showed no evidence of an immunocompromised state. Antiviral serological data, identification of the viral genome by polymerase chain reaction and serial spinal cord magnetic resonance imaging findings, supported the diagnosis of CMV associated TM in a non-immunocompromised patient. CONCLUSIONS: Our case further indicates that CMV infection should be included in the differential diagnosis of TM of uncertain etiology, in the immunocompetent patient. Clinical, immunological and neuroimaging findings indicate that post-infectious immune mediated inflammation, seems the most probable pathogenetic mechanism in this case. PMID- 11859443 TI - Vesicoureteral reflux and bladder management in spinal cord injury patients. PMID- 11859444 TI - [Atherosclerosis: immunological reply to the wrong address?]. PMID- 11859445 TI - [Long-term results after carotid reconstruction in patients with completed stroke caused by cerebrovascular occlusive disease]. AB - Long-term results after carotid reconstruction in patients with completed stroke caused by cerebrovascular occlusive disease. BACKGROUND AND OBJECTIVE: In patients with completed stroke caused by cerebrovascular occlusive disease the reconstruction of high grade carotid artery stenosis is discussed as prophylaxis for prevention of recurrent stroke. The operative complications of all patients who underwent surgery for carotid occlusive disease in the Heinrich Heine University between 1990 and 1999 were evaluated in this study. PATIENTS AND METHODS: We present the results of 596 reconstructions in 537 patients (404 men, 133 women; mean age 64,5 plus minus 9,4 years). The operative technique of choice was thromboendarterectomy of the carotid bifurcation with vein-patch closure. RESULTS: A postoperative ipsilateral neurological deficit was observed in 41 patients (6,9 %): 4,7 % permanent, 2,2 % transitory. Thirtyeight percent of the permanent deficits appeared after reconstructions performed within six weeks after stroke and only 9 % after surgery between six and twelve weeks. The in hospital mortality was 0,7 % for neurological and 1,0 % for cardiopulmonary complications. During long-term follow-up (mean follow-up time 50 plus minus 34 months) only 17 (3,7 %) of 462 patients, which could be evaluated, presented with a new neurological event for the reconstructed side. Fifty-eight percent had none or minimal neurological deficits. 115 Patients (24,0 %) died during follow-up. CONCLUSION: Even after completed stroke carotid reconstruction can be carried out successfully in selected patients with an acceptable complication rate in prophylaxis of recurrent stroke. In our retrospective study the optimal time for surgery seems to be between six and twelve weeks after stroke, but new imaging techniques may alter in the future the time window for operation in patients with an unstable neurological situation early after stroke. The reduction of social mobility and the high mortality in follow-up in correlation to patients without stroke are mostly related to accompanying severe cardiopulmonal risk factors. PMID- 11859446 TI - [Differences in insulin secretion facilitate the differential diagnosis of insulinoma and factitious hypoglycaemia]. AB - HISTORY: A 33-year-old female nurse (married; two children; BMI 30.9 kg/m2) had recurrent episodes of symptomatic hypoglycaemia over some months. INVESTIGATIONS: Two fasting tests were terminated after 26 hours because the patient became unconscious. Improved insulin/glucose ratio was infinity and 6.1 [mU/l]/[mg/dl] (normal value < 0.5). An hyperinsulinaemic-hypoglycaemic angle "clamp test" produced a C-peptide suppression to minimally 0.26 - 0.38 nmol/l (normal value 0.06 +/- 0.01 nmol). There was no spontaneous or paradoxical burst in insulin or C-peptide concentration after either the fasting or the "clamp test". Serum analysis of sulphonylurea on several occasions documented an increase of glibenclamide above therapeutic range. TREATMENT AND COURSE: The patient denied any intake of oral antidiabetic preparations, but there were no further hypoglycaemia attacks in subsequent months. DIAGNOSIS: The demonstration of sulphonylurea in serum confirmed the diagnosis of factitious hypoglycaemia. CONCLUSION: With regard to insulin or C-peptide suppression, the results of the fasting and clamp tests are the same in factitious hypoglycaemia and insulinoma. However, under the influence of sulphonylurea drugs there are no insulin or C peptide bursts so typical of insulinoma. In case of doubt, detection of sulphonylurea preparations in serum or urine is the only reliable way of diagnosing factitious hypoglaema due to the ingestion of sulphonylurea. PMID- 11859447 TI - [Coronary artery surgery in patients with diabetes mellitus]. PMID- 11859448 TI - [Sudeck's atrophy: pathophysiology and treatment of a complex pain syndrome]. AB - Sudeck's atrophy: pathophysiology and treatment of a complex pain syndrome. SUMMARY: The "Morbus Sudeck" or Complex Regional Pain Syndrome (CRPS) forms a typical triad of motor, sensory and autonomic symptoms. It is clinically characterized by spontaneous pain and hyperalgesia not limited to a single nerve territory and disproportionate to the inciting event. An underlying pathophysiology which could explain the whole symptomatology of CRPS is still unknown. Therefore, nowadays therapy is still symptomatic. However, recent research led to a better understanding of the disease and to the beginning of a pathophysiologically orientated therapy. PMID- 11859449 TI - [Immunopathogenesis of atherosclerosis: the Mainz hypothesis]. PMID- 11859450 TI - [Diagnosis-related groups as a medico-ethical challenge]. PMID- 11859451 TI - [Breast cancer in the man]. PMID- 11859452 TI - [Glitazones--a new class of drugs in treatment of type 2 diabetes]. PMID- 11859455 TI - Effects of naturally occurring isoflavones on prostaglandin E2 production. AB - Previously, we reported that the isoflavones tectorigenin and tectoridin, a glycosylated tectorigenin, isolated from the rhizomes of Belamcanda chinensis have an activity to inhibit prostaglandin (PG) E2 production in 12-O tetradecanoylphorbol 13-acetate (TPA)-stimulated rat peritoneal macrophages. The inhibitory effect of tectorigenin is more potent than that of tectoridin. In this study, we investigated the structure-activity relationship of various isoflavones in the inhibition of PGE2 production in TPA-stimulated rat peritoneal macrophages. The isoflavones examined were isolated from the rhizomes of B. chinensis, and the flowers and the rhizomes of Pueraria thunbergiana. The order of potency to inhibit PGE2 production was as follows; irisolidone, tectorigenin > genistein > tectoridin (tectorigenin 7-glucoside), glycitein > daidzein. Kakkalide (irisolidone 7-xylosylglucoside), glycitin (glycitein 7-glucoside), daidzin (daidzein 7-glucoside), puerarin (daizein 8-glucoside), and genistin (genistein 7-glucoside) showed no significant inhibition. These findings indicated that 6-methoxylation and 5-hydroxylation increase the potency to inhibit PGE2 production and 7-O-glycosylation decreases the inhibitory activity. PMID- 11859456 TI - Furanoligularenone, an eremophilane from Ligularia fischeri, inhibits the LPS induced production of nitric oxide and prostaglandin E2 in macrophage RAW264.7 cells. AB - Furanoligularenone (1), a known eremophilane, was identified from Ligularia fischeri (Compositae) together with 3-oxo-8alpha-hydroxy-10alphaH-eremophila 1,7(11)-dien-12,8beta-olide (2) and 3-oxo-8alpha-methoxy-10alphaH-eremophila 1,7(11)-dien-12,8beta-olide (3), by its potent inhibition of LPS-induced nitric oxide (NO) and prostaglandin E2 (PGE2) production in RAW 264.7 cells. Compound 1 also suppressed the expression of iNOS and COX-2 mRNA and protein in a dose dependent manner. Taken together, compound 1 inhibits the production of NO and PGE2 at the transcription of iNOS and COX-2 genes, and would be responsible for the anti-inflammatory activities of the Ligularia fischeri. PMID- 11859457 TI - Antiviral activity of seven iridoids, three saikosaponins and one phenylpropanoid glycoside extracted from Bupleurum rigidum and Scrophularia scorodonia. AB - As part of our screening of antiviral agents from medicinal plants, 11 compounds from plant origin (Bupleurum rigidum and Scrophularia scorodonia), three saikosaponins, seven iridoids and one phenylpropanoid glycoside were tested in vitro against herpes simplex type I (HSV-1), vesicular stomatitis virus (VSV) and poliovirus type 1. Five of these compounds showed antiviral activity against VSV. The percentages of cellular viability at the non-toxic limit concentrations of the active compounds were: verbascoside 53.6 % at 500 microg/ml, 8 acetylharpagide 32.1 % at 500 microg/ml, harpagoside 43.3 % at 450 microg/ml, scorodioside 47.8 % at 500 microg/ml and buddlejasaponin IV 56.9 % at 25 microg/ml. Although none of the saikosaponins were active against HSV-1, the iridoid scorodioside showed moderate in vitro anti-HSV-1 activity (30.6 % at 500 microg/ml). However, none of the compounds tested in this survey had any effect against poliovirus. PMID- 11859458 TI - In vitro anti-HIV activity of oleanolic acid on infected human mononuclear cells. AB - Oleanolic acid is a triterpenoid which is quite common in nature in the form either of free acid or in triterpenoid saponin glycosides. This study describes the effect of oleanolic acid on the growth of human immunodeficiency virus-1 (HIV 1) in cultures of human peripheral mononuclear cells (PBMC) and of monocyte/macrophages (M/M). Its inhibitory activity was also evaluated on PBMC obtained from HIV-1 infected patients. Results obtained show that oleanolic acid inhibits the HIV-1 replication in all the cellular systems used (EC50 values: 22.7 microM, 24.6 microM and 57.4 microM for in vitro infected PBMC, naturally infected PBMC and M/M, respectively). As regards the mechanism of action, oleanolic acid inhibits in vitro the HIV-1 protease activity. PMID- 11859459 TI - In vitro modulation of telomerase activity, telomere length and cell cycle in MKN45 cells by verbascoside. AB - Screening of natural products with anti-tumor activity as telomerase inhibitor is a new subject in the field of tumor therapy. Using telomerase PCR ELISA, telomere DNA hybridization and flow cytometry analysis, the effects of verbascoside, a phenylpropanoid glucoside extracted from Pedicularis striata Pall, on telomerase activity, telomere length and cell cycle of human gastric carcinoma cells MKN45 was examined in vitro. After being treated with a 50 % inhibition concentration of verbascoside (17.8 microg/ml), telomerase activity in the cells was significantly inhibited but not in the cellular supernatant, the average telomere length became remarkably short, and the sub-G0 /G1 peak and G2/M arrest were also displayed when compared to the control cells. These results suggest that verbascoside mediated-cell differentiation and apoptosis may be affected by telomere-telomerase-cell cycle dependent modulation. Thus, the antitumor mechanism of verbascoside is demonstrated once more by its inhibiting effect on telomerase activity in tumor cells, and the telomerase assay may provide a valuable screening method for antitumor activity of natural products. PMID- 11859460 TI - Inhibitory effect of tumor cell proliferation and induction of G2/M cell cycle arrest by panaxytriol. AB - Panaxytriol, a polyacetylenic compound, isolated from red ginseng (Panax ginseng C.A. Meyer), was studied to determine its effects on the growth and cell cycle of tumor cell lines. The compound showed both significant cytotoxicity and inhibition of DNA syntheses in various tumor cells tested. For P388D1, a mouse lymphoma cell line, IC50 values for cytotoxicity and inhibition of DNA synthesis were 3.1 and 0.7 microg/ml, respectively. The cytotoxic effect of panaxytriol was both time- and dose-dependent. It also induced the cell cycle arrest of P388D1 at the G2/M phase, which was measured through flow cytometry. Particularly, the proportion of cells in the G2/M phase of the cell cycle increased from 9 % to 26 and 48 %, respectively, after 24 and 36 h exposure to panaxytriol at 5 microg/ml. There were corresponding decreases in the proportion of cells at the G0/G1 phase. The S phase also decreased during the 36-h treatment. PMID- 11859461 TI - Induction of apoptosis by 3,4'-dimethoxy-5-hydroxystilbene in human promyeloid leukemic HL-60 cells. AB - 3, 4'-Dimethoxy-5-hydroxystilbene (DMHS) is a hydroxystilbene compound obtained by methylation and acid hydrolysis of piceid (resveratrol-3-O-glucoside) from Polygonum cuspidatum. Herein, we report that DMHS induces programmed cell death or apoptosis in human promyelocytic leukemic HL-60 cells. We found that treatment of HL-60 cells with DMHS suppressed the cell growth in a concentration-dependent manner with an IC50 value of 25 microM. DMHS increased internucleosomal DNA fragmentation in a time-dependent manner. The cell death by DMHS was partially prevented by the caspase inhibitor, zVAD-fmk. DMHS caused activation of caspases such as caspase-3, -8, and -9. Immunoblot experiments revealed that DMHS-induced apoptosis was associated with the induction of Bax expression. The release of cytochrome c from mitochondria into the cytosol was increased in response to DMHS. Taken together, our present results indicated that DMHS leads to apoptotic cell death in HL-60 cells through increased Bax expression and release of cytochrome c into cytosol and may be considered as a good candidate for a cancer chemopreventive agent in humans. PMID- 11859462 TI - Different effects of baicalein, baicalin and wogonin on mitochondrial function, glutathione content and cell cycle progression in human hepatoma cell lines. AB - The effects of the flavonoids from Scutellaria baicalensis Georgi (baicalein, baicalin and wogonin) in cultured human hepatoma cells (Hep G2, Hep 3B and SK Hep1) were compared by MTT assay and flow cytometry. All three flavonoids dose dependently decreased the cell viabilities accompanying the collapse of mitochondrial membrane potential and the depletion of glutathione content. However, the influence of baicalein, baicalin or wogonin on cell cycle progression was different. All three flavonoids resulted in prominent increase of G2/M population in Hep G2 cells, whereas an accumulation of sub G1 (hypoploid) peak in Hep 3B cells was observed. In SK-Hep1 cells, baicalein and baicalin resulted in dramatic boost in hypoploid peak, but wogonin made mainly in G1 phase accumulation. These data, together with the previous findings in other hepatoma cell lines, suggest that baicalein, baicalin and wogonin might be effective candidates for inducing apoptosis or inhibiting proliferation in various human hepatoma cell lines. PMID- 11859463 TI - Involvement of protein kinase C and Na+/K+-ATPase in the contractile response induced by myricetin in rat isolated aorta. AB - The role of PKC and Na+/K+-ATPase in the vascular smooth muscle responses induced by the bioflavonoid myricetin was investigated. KCl induced a concentration dependent relaxation in arteries exposed to K+-free solution that was mainly mediated by an activation of Na+/K+-ATPase. Myricetin (50 microM) partially inhibited this vasorelaxant effect induced by KCl in intact rings, being unaffected in the endothelium-denuded rings. This inhibitory effect induced by myricetin was suppressed by the PGH2-TXA2 receptor antagonist, SQ 29,548, and the PKC inhibitor, staurosporine. Myricetin also induced an endothelium-dependent contractile response which was increased in the presence of PMA and reduced by staurosporine. In conclusion, myricetin both modulates Na+/K+-ATPase-induced vasodilatation acting as a functional inhibitor of Na+/K+-ATPase activity and activates protein kinases, including PKC, to induce contraction. These effects appear to be related to the activation of PGH2-TXA2 receptors on vascular smooth muscle by the TXA2 released from endothelium.NA:noradrenalineNA+/K+-ATPase pump:sodium-potassium-activated ATPasePKC:protein kinase CPMA:phorbol 12 myristate 13-acetateTXA2:thromboxane A2The role of PKC and Na+/K+-ATPase in the vascular smooth muscle responses induced by the bioflavonoid myricetin was investigated. KCl induced a concentration-dependent relaxation in arteries exposed to K+-free solution that was mainly mediated by an activation of Na+/K+ ATPase. Myricetin (50 microM) partially inhibited this vasorelaxant effect induced by KCl in intact rings, being unaffected in the endothelium-denuded rings. This inhibitory effect induced by myricetin was suppressed by the PGH2 TXA2 receptor antagonist, SQ 29,548, and the PKC inhibitor, staurosporine. Myricetin also induced an endothelium-dependent contractile response which was increased in the presence of PMA and reduced by staurosporine. In conclusion, myricetin both modulates Na+/K+-ATPase-induced vasodilatation acting as a functional inhibitor of Na+/K+-ATPase activity and activates protein kinases, including PKC, to induce contraction. These effects appear to be related to the activation of PGH2-TXA2 receptors on vascular smooth muscle by the TXA2 released from endothelium. PMID- 11859464 TI - Marked decrease of cyclosporin absorption caused by phellamurin in rats. AB - Phellamurin is a flavonoid glycoside that is abundant in the leaves of Phellodendron wilsonii Hayata et Kanehira (Rutaceae). In vitro everted rat intestine study indicated that phellamurin inhibited intestinal P-glycoprotein in a dose-dependent manner. In order to investigate the effect of phellamurin on cyclosporin absorption and disposition, rats were given cyclosporin (5 mg/kg) with or without phellamurin in a parallel design. Fluorescence polarization immunoassay was used to determine the blood concentration of cyclosporin. Unanticipatedly, our results indicated that the coadministration of phellamurin significantly decreased the Cmax of cyclosporin by 77 % and reduced the AUC(0 infinity) of cyclosporin by 56 %. This indicated that a serious interaction occurred between phellamurin with cyclosporin. To ensure the efficacy of cyclosporin, we suggest that the coadministration of phellamurin or Phellodendron wilsonii with cyclosporin should be avoided. PMID- 11859465 TI - Cytotoxic butanolides and secobutanolides from the stem wood of Formosan Lindera communis. AB - Bioassay-guided fractionation of the chloroform-soluble fraction from the stem wood of Formosan Lindera communis led to the isolation of two new cytotoxic butanolides, lincomolide C (1), lincomolide D (2), as well as two new secobutanolides, secolincomolide A (3), secolincomolide B (4), together with four known compounds, including one butanolide, isoobtusilactone A (5), and one secobutanolide, secoisolancifolide (6), one sesquiterpene, epi-cubenol (7), one steroid: beta-sitosterol (8). Compound 4 existed in enol [(2E)-2-[(1R)-1-hydroxy 2-oxo-propyl]-6,9-dimethyl-8-hydroxy-hexadec-2,7-dienoic acid methyl ester] (4a) and keto [(2E)-2-[(1R)-1-hydroxy-2-oxo-propyl]-6,9-dimethyl-8-oxo-hexadec-2-enoic acid methyl ester] (4b) tautomers. Compound 2 showed significant cytotoxicity against P-388 and HT-29 cancer cell lines. Also, 2 exhibited marginal activity against A549, 5 against P-388, and 1 against P-388, A549 and HT-29 cancer cell lines. Their structures were determined by spectroscopic analyses. PMID- 11859466 TI - Cytotoxic coumaronochromones from the roots of Euchresta formosana. AB - Five new coumaronochromones, euchretins J-N (1 - 5), along with twelve known compounds, euchretins A (6), C, D, F, H, I, (+)-matrine, (-)-cytisine, quercetin, trifolirhizin, retusin 8-methyl ether, and genistein, were isolated from the methanolic extracts of Euchresta formosana. The structures of compounds 1 - 5 were established by spectroscopic analyses. Among them, compounds 1, 4 and 6 were demonstrated to have cytotoxicity against 59T cell line, and compound 1 was also found to be active against SCM-1 cell line. PMID- 11859467 TI - Quantitative determination of the dual COX-2/5-LOX inhibitor tryptanthrin in Isatis tinctoria by ESI-LC-MS. AB - Isatis tinctoria L. is an old European and Chinese dye plant and anti inflammatory herb from which the potent cyclooxygenase-2 and 5-lipoxygenase inhibitor tryptanthrin (1) (indolo-[2,1-b]-quinazoline-6,12-dione) was recently isolated as one of the active principles. An HPLC method for the quantitative analysis of the compound in plant material was developed. Reproducible extraction was achieved by accelerated solvent extraction (ASE). Detection by UV at 254 and 387 nm and by electrospray-MS were compared. The low tryptanthrin content in the herb and possible interferences required isocratic high-performance liquid chromatography coupled with electrospray-MS in single ion mode. More than 70 Isatis samples of different origin were analyzed. The tryptanthrin content in leaf samples varied from 0.56 to 16.74 x 10(-3) %. PMID- 11859468 TI - Determination of ginsenosides from Panax ginseng using micellar electrokinetic chromatography. AB - A method for determination of ginsenosides in herbal medicinal products (HMPs) was developed using micellar electrokinetic chromatography (MEKC). Within 22 minutes 7 major ginsenosides were well separated. In order to demonstrate the accuracy, precision and robustness for the main target analyte the method was exemplarily validated for the determination of Rb1 according to ICH guidelines. Compared to chromatographic analysis, several benefits of capillary electrophoresis (CE) could be demonstrated such as high separation efficiency in an aqueous buffer without any organic solvent and shorter run time per assay. PMID- 11859469 TI - Inhibitors of 5alpha -reductase type I in LNCaP cells from the roots of Angelica koreana. AB - A prenylated coumarin, osthenol (1) and a sesquiterpene, bisabolangelone (2) have been isolated as active principles with 5alpha-reductase type I inhibitory effects in LNCaP cells from the roots of Angelica koreana Max. by bioassay-guided chromatographic fractionation. Osthenol exhibited a highly potent inhibitory activity on 5alpha-reductase type I in LNCaP cells with an IC50 value of 0.1 microg/ml, which is about 200 times more potent than the positive control, finasteride (IC50 = 19.8 microg/ml). Bisabolangelone also inhibited the activity of 5alpha-reductase type I in LNCaP cells (IC50 = 11.6 microg/ml), indicating that these compounds are possible candidates for the development of new drugs to treat human endocrine disorders associated with overproduction of DHT by 5 alpha reductase type I. In addition, four compounds isooxypeucedanin, oxypeucedanin hydrate, oxypeucedanin and isoimperatorin were also isolated and found to be inactive in the 5alpha-reductase assay systems used in the present study. PMID- 11859470 TI - Composition and antifungal activity of the essential oil of Solidago chilensis. AB - Volatile constituents of the essential oils from leaves and inflorescences of Solidago chilensis Meyen were analyzed by GC-FID, GC-MS and 13C-NMR and thirty six different compounds were identified. Pumiloxide, an unusual labdane diterpene, was found to be one of the major components in both oils (15.3 % and 12.3 %, respectively). Other important constituents were limonene and several sesquiterpenes, mainly gamma-cadinene. The antifungal activity of the leaf oil was assayed against five different strains of filamentous fungi and one yeast. Paper disk agar diffusion test showed human pathogenic dermatophytes to be the most sensitive. PMID- 11859471 TI - Antiplasmodial activity of Uvaria klaineana. AB - Crude extracts of Uvaria klaineana Engler and Diels (Annonaceae) stems showed in vitro activity against chloroquine-resistant K1 strain of Plasmodium falciparum. The most active extract was the basic dichloromethane extract containing crude alkaloids (IC50 = 3.55 microg/mL). The bioassay-guided fractionation of this extract led to the isolation of the major alkaloid crotsparine (1) which showed an antiplasmodial activity against the chloroquine-sensitive Thai strain of P. falciparum and the chloroquine-resistant K1 and FcB1 strains of P. falciparum. Two minor alkaloids were also identified as crotonosine (2) and zenkerine (3). Their structures were elucidated using 2D-NMR techniques. PMID- 11859472 TI - Nematocidal activity of isoquinoline alkaloids against a species of diplogastridae. AB - We examined the nematocidal activity of various isoquinoline alkaloids. The method of evaluating nematocidal activity involved the microscopic examination of the state of larvae after the compound was administered to determine the minimum lethal concentration (MLC). In addition, cytotoxic activity (IC50) was examined using HL-60 culture cells. The compound dehydrocorydaline was found to have considerable nematocidal activity and weak cytotoxicity; its cytotoxicity value was equal to that of thiabendazole. We conclude that dehydrocorydaline, with its attractive balance between nematocidal activity and weak cytotoxicity, is the most promising of the tested compounds as a possible remedy for parasitic diseases. PMID- 11859473 TI - Phototoxic and apoptosis-inducing capacity of pseudohypericin. AB - Pseudohypericin (PH) and hypericin (H) are photosensitizing plant pigments from Hypericum perforatum. H displays cytotoxic and apoptosis-inducing effects in neoplastic cell lines. Here, we have assessed the phototoxic and apoptosis inducing capacity of PH compared to H in a cell culture model with human leukemic lymphoma cells (Jurkat). Treatment with both photoactivated H and PH resulted in a dose-dependent inhibition of cell proliferation, whereas not photoactivated H and PH had no effect at the concentrations tested. The half-maximal inhibitory concentration (IC50) of H was lower (100 ng/mL) as compared to PH (200 ng/mL) (p < 0.05). In an apoptosis assay we found a dose-dependent increase of DNA fragmentation after treatment with both photoactivated H and PH. The cytotoxic potential of PH should be taken into account during systemic therapy with Hypericum extracts, since PH is about two times more abundant than H in Hypericum perforatum. PMID- 11859474 TI - Ellagitannins from Lagerstroemia speciosa as activators of glucose transport in fat cells. AB - Glucose transport enhancers were searched for in Lagerstroemia speciosa, a Philippine local herbal medicine used for diabetes mellitus. Bioassay-guided fractionation of the aqueous acetone extract of the leaves afforded three active ellagitannins, lagerstroemin, flosin B and reginin A, identified by NMR and optical rotation. These compounds increased glucose uptake of rat adipocytes, and could be responsible for lowering the blood glucose level. PMID- 11859475 TI - Aesculin possesses potent hypouricemic action in rodents but is devoid of xanthine oxidase/dehydrogenase inhibitory activity. AB - The natural product aesculin was demonstrated to possess potent hypouricemic effects in in vivo models of hyperuricemia in both mice and rats pretreated with oxonate. Aesculin, when administered intraperitoneally to the oxonate-induced hyperuricemic rodents, was able to elicit dose-dependent hypouricemic effects. At doses of 150 mg/kg of aesculin or above, the serum urate levels of the oxonate pretreated mice were not different from normal mice. Such an effect in mice was observed as quick as 1.5 h after aesculin administration and was persistent for at least 5 h after aesculin administration. In rats, similar hypouricemic effects of intraperitoneally administered aesculin could also be demonstrated at doses of 100 mg/kg of aesculin or above, the serum urate levels of the oxonate-pretreated rats were not different from normal rats. Again, the effect persisted for at least 5 h after aesculin administration. In both rodents, however, oral administration at the same doses did not produce any observable hypouricemic effects. In addition, aesculin, when tested in vitro on rat and mouse liver homogenates, did not elicit any measurable inhibitory actions on the xanthine oxidase/xanthine dehydrogenase activities. PMID- 11859476 TI - The influence of the harvest cut height on the quality of the herbal drugs Melissae folium and Melissae herba. AB - Variability in both the content and quality of essential oil was observed in herb and leaf drugs in dependence on the harvest cut height of lemon balm (Melissa officinalis L. cv. Citra). Three different cuts were carried out on the respective plants. The oil content in the herb was highest in the top third (0.13 % V/m), satisfactory in the herb including both the top and middle thirds (0.08 % V/m) and lowest in the whole herb (0.06 % V/m). The oil content in the leaves of the respective herbs was in the range 0.39 % - 0.14 % V/m (top third part - whole aerial part). The percentage of citrals, linalool and beta-caryophyllene in essential oil decreased in the basipetal direction, whilst the amounts of beta caryophyllene oxide and citronellal increased in the same direction. Citrals (A and B): 55.79 % in the top third part of the herb, 48.46 % in the whole herb, 59.74 % and 56.87 % in the leaves from that parts, respectively. Similarly, beta caryophyllene: 5.01 %, 3.87 %, 6.97 %, 5.13 %; beta-caryophyllene oxide: 17.19 %, 24.07 %, 15.64 %, 17.82 %; citronellal: 2.73 %, 5.51 %, 2.82 %, 6.44 %. PMID- 11859477 TI - Three new prenylated isoflavonoids from the root bark of Erythrina vogelii. AB - Phytochemical investigation of the CH2Cl2 extract of Erythrina vogelii led to the isolation of five isoflavonoids. Three prenylated isoflavonoids are new natural compounds. The isolation of the antifungal compounds was monitored by inhibition of the growth of Cladosporium cucumerinum in a direct TLC bioautographic assay. The structures of these compounds were elucidated by spectroscopic techniques. PMID- 11859478 TI - New lupane glycosides from Pulsatilla chinensis. AB - Two new lupane glycosides along with five known triterpenoids were isolated from the roots of Pulsatilla chinensis (Ranunculaceae). The structures of the new glycosides were determined to be 3-O-beta-D-glucopyranosyl(1-->3)-alpha-L arabinopyranosyl-23-hydroxybetulinic acid 28-O-alpha-L-rhamnopyranosyl(1-->4) beta-D-glucopyranosyl(1-->6)-beta-D-glucopyranosyl ester (pulsatilloside D, 6) and 3-O-[beta-D-glucopyranosyl(1-->4)][alpha-L-rhamnopyranosyl(1-->2)]-alpha-L arabinopyranosyl-23-hydroxybetulinic acid 28-O-alpha-L-rhamnopyranosyl(1-->4) beta-D-glucopyranosyl(1-->6)-beta-D-glucopyranosyl ester (pulsatilloside E, 7) by spectroscopic analysis and chemical methods. The compounds were evaluated for cytotoxic activities against K-562 human leukemia and HeLa cells. PMID- 11859479 TI - Constituents of the leaves and twigs of Ficus hispida. AB - A new norisoprenoid, ficustriol (1), and the known phenanthroindolizidine alkaloid O-methyltylophorinidine (2), were isolated from a CHCl3 extract of the leaves and twigs of Ficus hispida. O-Methyltylophorinidine showed potent cytotoxic activity when tested against a small panel of human cancer cells, while ficustriol was inactive. The structure and stereochemistry of 1 were determined using chemical and spectral methods. PMID- 11859480 TI - New lycopodium alkaloids from Huperzia serrata. AB - Three new lycopodium alkaloids, 11alpha-hydroxyphlegmariurine B (2), 7alpha hydroxyphlegmariurine B (3) and 7alpha,11alpha-dihydroxyphlegmariurine B (4) along with a known compound, phlegmariurine B (1), were isolated from the herb Huperzia serrata. The structures of the above compounds were elucidated based on spectroscopic studies. PMID- 11859481 TI - Authentication of stems of Dendrobium officinale by rDNA ITS region sequences. AB - The rDNA ITS regions of five Dendrobium species were sequenced. Each Dendrobium species was found to have a unique sequence in the ITS region, so that they could be easily distinguished at the DNA level. The aligned 644 bp of the ITS region includes 235 bp ITS1, 163 bp 5.8S, and 246 bp ITS2. One hundred and eighty-nine sites are variable. The sequences of D. officinale could be easily distinguished from the other four adulterant species according to the sequence variation at 11 sites, 7 in ITS1, 1 in 5.8S, and 3 in ITS2. These could be used as molecular characters to distinguish the stems of D. officinale from the adulterants. PMID- 11859483 TI - Top 10 reasons a manuscript is rejected. PMID- 11859482 TI - Evolutionary fate of an unstable human minisatellite deduced from sperm-mutation spectra of individual alleles. AB - Although mutation processes at some human minisatellites have been extensively characterized, the evolutionary fate of these unstable loci is unknown. Minisatellite instability is largely germline specific, with mutation rates up to several percent and with expansion events predominating over contractions. Using allele-specific small-pool polymerase chain reaction, we have determined sperm mutation spectra of individual alleles of the highly unstable human minisatellite CEB1 (i.e., D2S90). We show that, as allele size increases, the proportion of contractions rises from <5% to 50%, with the average size of deletion increasing and eventually exceeding the average size of expansion. The expected net effect of these trends after many generations is an equilibrium distribution of allele sizes, and allele-frequency data suggest that this equilibrium state has been reached in some contemporary human populations. PMID- 11859484 TI - Nursing nomenclatures and eye-roll anxiety control. PMID- 11859485 TI - Preserving professional values. PMID- 11859486 TI - Time for a concerted effort to strengthen nursing and midwifery services globally. PMID- 11859487 TI - Continuing the case for the baccalaureate. PMID- 11859488 TI - Improving faculty publication output: the role of a writing coach. AB - Publishing academic papers is recognized by faculty as vital not only to their careers, but also to the standing of their school within the university and the discipline. Although writing is perceived as a critical, high-priority task, it often has low follow-through behavior. To facilitate the publication output of our faculty, a part-time writing coach was hired. Blanchard's situational leadership II model (1985), which tailors leadership style to the needs of the group, indicated a framework of coaching and support would best meet faculty writing needs. The literature further suggested that an ongoing coaching relationship in the form of a committed partnership would empower faculty to achieve beyond prior performance. We present a model to show how coaching and support facilitate perseverance in writing for publication. Sixteen of 26 faculty sought the coach's help in the first 2.5 years, generating 21 submissions to peer reviewed journals. Fifteen of these were published or accepted (71 per cent), and one was still under review when this article was written. Five rejected papers had not yet been resubmitted. Faculty with recent doctorates appeared to benefit most from coaching. PMID- 11859489 TI - Research support by doctoral-granting colleges/schools of nursing. AB - Colleges and schools of nursing with doctoral programs focus on developing quality research programs. One effective way of managing and nurturing a research program is through the implementation of a nursing research office or center. The purpose of this study is to describe the resources provided by the colleges/schools of nursing with doctoral programs for research development. A self-report questionnaire, developed by the research team, was mailed to all schools of nursing offering doctoral programs. The response rate was 79 per cent (65/82 schools). Results indicated that 56 schools (86.2 per cent) have designated research support offices. The main goals of nursing research offices are to increase the amount of extramural funding and to promote dissemination of scholarly work via publications and presentations. The majority of research offices provide assistance with grants and the research process and offer educational programs. Most doctoral-granting schools are providing some support for research activities. However, the degree of investment in research support varied widely among the responding schools. This study suggests that it takes both time and institutional commitment to build a successful research environment. Although necessary for research development, support services are not sufficient by themselves. Instead, they need to be considered in the light of individual (e.g., faculty interest and motivation) and group (e.g., culture of scholarship) factors within each school. PMID- 11859490 TI - Evaluating nurse competency: evidence of validity for a skills recredentialing program. AB - A multitrait, multimethod approach was used to evaluate the validity of a hospital-based nurse competency assessment program as a mechanism for assuring compliance with the three standards for competence established by the National Council of State Boards of Nursing, Inc. For selected competencies, both knowledge and skill performance were evaluated under simulated conditions and under routine patient care conditions. Under simulated conditions, pass rates on first attempts among the 368 participating nurses were high as expected (82 per cent to 94 per cent on knowledge and 75 per cent to 99 per cent on performance). Offering immediate educational feedback followed by second attempts yielded a 97 per cent overall pass rate. Competencies assessed under both simulated and patient care conditions (35 sets of observations) revealed no differences in assessed competence. The equivalence of assessments made under simulated and patient care conditions supports the validity of competency assessment programs currently in use in many large health care settings. PMID- 11859491 TI - Using standardized patients in advanced practice nursing education. AB - Standardized patient encounters, a pedagogic approach to helping students develop appropriate clinical skills, are widely used in medical education and are being adopted by advanced practice nursing programs. Two types, simulated clinical encounters, in which students complete an episodic or comprehensive visit, and objective structured clinical experiences, multiple stations each presenting a different clinical problem, are used for formative and summative evaluation. Graduating adult and pediatric nurse practitioner students (N = 26) completed a simulated clinical encounter for an episodic visit. Students' performance on the simulated clinical encounter did not reflect their performance on other clinical evaluation measures or their performance on national certifying examinations. This one-time simulated clinical encounter was shown to lack the necessary reliability and validity to appropriately evaluate the clinical skills of nurse practitioner students. Videotaped, simulated patient encounters are useful, however as formative learning experiences. This reinforced our position that multiple site visits by faculty to students and their preceptors at all their clinical sites are necessary to accurately evaluate students' clinical performance. PMID- 11859492 TI - Nurse practitioners and physician assistants revisited: do their practice patterns differ in ambulatory care? AB - The education and regulation of nurse practitioners and physician assistants would suggest unique role differentiations and practice functions between the professions. This study explored to what extent their practice patterns in primary care actually differ. It was hypothesized that the primary care services provided by nurse practitioners would tend to be women and family health services, health prevention and promotion oriented, provided to minority and socioeconomic disadvantaged patients, and less dependent on physician supervision. In contrast, the services provided by physician assistants would more likely be medical/surgical oriented; diagnostic, procedural, and technical in nature; likely to be in rural areas; and more dependent on physician supervision. The study used patient data from the National Ambulatory Medical Care Survey and National Hospital Ambulatory Medical Care Survey. Although some differences emerged, the argument is not compelling to suggest strong, unique, practice differences across all ambulatory care settings between the two types of nonphysician providers. It is the specific type of ambulatory setting that influences the practice pattern for both provider groups. If practice patterns are less distinctive than previously believed, more opportunities for interdisciplinary education need to be explored, and health policies that promote a discipline-specific primary care workforce may need to be reexamined. PMID- 11859493 TI - Making connections: linking generalist and specialist essentials in baccalaureate community/public health nursing education and practice. AB - Nurse educators and potential employers of baccalaureate nursing graduates are challenged by the constant change in health care today. This change mandates competencies for professional nursing practice that preserves the relevant and useful skills and knowledge of the past, yet incorporates skills and knowledge for practice today and in the future. As in previous eras of health system change, identifying essential knowledge and competencies for baccalaureate graduates must begin by collaboration between nursing education and nursing service. This article reports the work of a task force of nurse educators whose charge was to revise "The Essentials of Baccalaureate Nursing Education for Entry Level Community/Public Health Nursing". An overview of their experience is presented with the aim of contributing to current best thinking between one nursing specialty organization and the American Association of Colleges of Nursing. PMID- 11859494 TI - [Cardiac cachexia]. PMID- 11859495 TI - [Surgical treatment of breast cancer being renewed]. PMID- 11859496 TI - [Positional vertigo caused by horizontal semicircular canalolithiasis--an underdiagnosed disorder]. PMID- 11859497 TI - [New knowledge about the role of mast cells in the immune defense]. PMID- 11859498 TI - [Aseptic necrosis of both femurs in a man with panniculitis]. PMID- 11859499 TI - [How useful is it to determine antibodies against Helicobacter pylori?]. PMID- 11859500 TI - [The use of cyclosporin A in the treatment of autoimmune disorders]. PMID- 11859501 TI - [What can the physician do in order to improve the traffic safety?]. PMID- 11859502 TI - [Sick elderly in the traffic--an increasing risk?]. PMID- 11859503 TI - [Alcohol and drugs in the traffic]. PMID- 11859504 TI - [Regulations for controlling fitness to drive]. PMID- 11859505 TI - [Problems with alertness and perception as causes for traffic accidents]. PMID- 11859506 TI - [Reducing injuries by improving safety structures in cars]. PMID- 11859507 TI - [How does fetus cheat mother's T-cells?]. PMID- 11859508 TI - [Is cancer spreading during laparoscopy?]. PMID- 11859509 TI - [Prevention of osteoporotic fractures]. PMID- 11859510 TI - [Views of the pathogenesis in Alzheimer's disease are changing]. PMID- 11859511 TI - [Dry cloning]. PMID- 11859512 TI - [Cancer immunotherapy]. PMID- 11859513 TI - [Eclampsia in Finland 1990-94]. PMID- 11859514 TI - [Bilateral subclavian steal syndrome--a rare cause of fainting]. PMID- 11859516 TI - [Treatment of pulmonary hypertension]. PMID- 11859515 TI - [Patients ask for sildenafil--how to treat?]. PMID- 11859517 TI - [Acetylsalicylic acid and the prevention of cancer]. PMID- 11859518 TI - [Cyclo-oxygenase 2 in cancer]. PMID- 11859519 TI - [Genes, HDL cholesterol and risk of coronary artery disease]. PMID- 11859520 TI - [Treatment of spasticity]. PMID- 11859521 TI - [Physicians' home visits--hopes and reality]. PMID- 11859522 TI - [When the abdomen is bloating]. PMID- 11859523 TI - [Involuntary weight loss]. PMID- 11859524 TI - [Essential hypertension--do we still not know its cause?]. PMID- 11859525 TI - [Should elevated systolic blood pressure be treated?]. PMID- 11859526 TI - [Therapy of hypertension--have we reached the goals?]. PMID- 11859527 TI - [Measuring the blood pressure: in the doctor's office, at home or during 24 hours with the machine?]. PMID- 11859528 TI - [When is blood pressure lowering needed?]. PMID- 11859529 TI - [Non-pharmacological treatment of hypertension]. PMID- 11859530 TI - [Pharmacological treatment of hypertension]. PMID- 11859531 TI - [Treatment and follow-up of a hypertensive patient]. PMID- 11859532 TI - Feeling your pain. Hospitals have to treat agony. But now they must find the best measure of it. PMID- 11859533 TI - On the edge on campus. The state of college students' mental health continues to decline. What's the solution? PMID- 11859534 TI - An unfair ticket to college. PMID- 11859535 TI - Body of evidence. PMID- 11859536 TI - Control of prescription costs--is it possible? AB - The author considers two choices for controlling the cost of prescription drugs: Return the prescription drug program to the TPA, or implement plan design changes. Making changes to the buying habits of the public is key. PMID- 11859537 TI - Depression's surprising toll on worker productivity. AB - Employers need to adopt a broad set of measures to combat depression, which has a serious impact on health care costs and productivity. They should start by conducting an assessment of the various ways depression surfaces and prepare carefully designed responses, involving HR, the EAP, line managers/supervisors, health plan vendors, managed behavioral firms, disability program managers and PBMs, among others. With such an approach, ultimately employers should realize improved medical plan costs, reduced disability expenditures, lower absence rates and enhanced productivity. PMID- 11859538 TI - How to achieve an ROI on your health care dollars. AB - Offering employees self-care information, which encourages them to decide what to do for themselves with and without provider assistance, can be a low-cost, effective approach to health care cost management. Components of such a program may include a printed self-care guide, workshops, software and a nurse advice line. Self-care is a process, not an event and, to be effective, a self-care program needs to be reinforced by ongoing communication. PMID- 11859539 TI - Managing overlapping federal FMLA and state leave regulations. AB - The overlap between the Family and Medical Leave Act and state leave laws can create complications. Employers, particularly those with multistate operations, may wish to consider an outsourced absence management system, which can remove the burden of day-to-day administration and the need to stay abreast of changing state regulations. However, employers cannot outsource the responsibility to set broad policy toward absence management. PMID- 11859540 TI - Affordable defined contribution health plans can help employers get through challenging times. AB - Defined contribution health plans, modeled on defined contribution retirement plans, seem to be the wave of the future. At a time when employers and employees alike are trying to contain their health care costs, and employers fight to retain their workers, defined contribution health plans can provide solutions for both employers and employees. The health purchasing alliance model has met with success in both the small and mid-size market. PMID- 11859541 TI - Sanctions with bite could affect hospitals. Final IRS rule might mean more aggressive enforcement, but it includes protections, too. PMID- 11859542 TI - Blues probes cost of care. PMID- 11859543 TI - Shelter for whistleblowers. Appeals court reinstates False Claims Act lawsuit. PMID- 11859544 TI - AHA to take lead on IT. Catalyst role may speed pace of standardization. PMID- 11859545 TI - Boston's CareGroup may dismantle system. PMID- 11859546 TI - Critical-care push intensive. PMID- 11859547 TI - NSLIJ (North-Shore Long Island Jewish Health System) fosters education. PMID- 11859548 TI - Post-acute firms see slow recovery. PMID- 11859549 TI - New lease on life: device re-use gets legitimized. Illinois state senator led push to regulate 'secret' industry of reconditioned devices. PMID- 11859550 TI - The privatization of quality. Washington seems to be hoping that the medical errors issue will just go away. PMID- 11859551 TI - The hard lessons of September. PMID- 11859552 TI - High on tech, low on budget. Annual survey spotlights growing gap between what execs want from information technology and what they'll spend to achieve it. PMID- 11859553 TI - Straight talk: new approaches in healthcare. Clinical information systems: where are we today, where do we need to be, and how do we get there? PMID- 11859554 TI - Absolute minimum. California announces its decision on nurse-staffing ratios, and other states watch and wait for the ripples to come their way. AB - After two years of wrangling, California hospitals learned how many patients the state government believes one nurse should care for safely. The announcement last week could well reverberate nationally. The ratio of one nurse to six patients in medical-surgical units is twice the number of patients supported by the state's nurse association and four fewer than that supported by the California Healthcare Association. PMID- 11859555 TI - Cutting through borders and barriers. Northern Indiana alliance of providers overcomes political, technological obstacles in effort to develop to shared medical record. PMID- 11859557 TI - Therapeutic play and the impact on anxiety in hospitalized children. AB - Zahr's (1998) study supported the use of puppet shows, as therapeutic play, to decrease anxiety in hospitalized preschoolers. The results of this study support our group's research utilization project to educate nurses on the effects of therapeutic play on anxiety levels in hospitalized children. Nurses could use this information to implement therapeutic play in hospitals throughout the world. Feasibility issues would include the cost of materials needed and the time involved to educate nurses. More research should be done on therapeutic play as a method to decrease anxiety in children. Future research could be conducted using other cultures or age groups. PMID- 11859556 TI - Awareness of choices regarding prostate cancer screening. PMID- 11859558 TI - Intercessory prayer and its effect on patients with rheumatoid arthritis. AB - Matthews et al's. (2000) study suggested that in person intercessory prayer was useful in the medical treatment of patients with RA, improving overall health. However, distant prayer showed no overall improvement. This study could be used to support a research utilization project to educate nurses on the potential benefits of prayer on the healthcare and health outcomes of patients with certain illnesses. An in-service could be made available for nurses to educate them on potential benefits of prayer. A feasibility issue would be cost. Religion and spiritual beliefs can influence a patient's level of health and self-care behaviors. It is essential for nurses to meet the spiritual needs of their patients. A nurse should also recognize that there are many different religions and not all patients have the same religion and beliefs. PMID- 11859559 TI - Attention all nurses and nursing students. PMID- 11859560 TI - Human genome project: past, present, future. PMID- 11859561 TI - In search of new disease models in the mouse using ENU mutagenesis. PMID- 11859563 TI - Lessons from the sequence of human chromosome 22. PMID- 11859562 TI - Genome-wide comparisons between human and tetraodon. PMID- 11859564 TI - ATM: from phenotype to functional genomics--and back. PMID- 11859565 TI - The genetics of deafness: a model for genomic and biological complexity. PMID- 11859566 TI - Incontinentia pigmenti: the first single gene disorder due to disrupted NF-kappa B function. PMID- 11859567 TI - The human genome. Biology and medicine. Introduction. PMID- 11859568 TI - [Long-time results of ITI full-screw implants. Analysis of 10-year effectiveness of 468 implants]. PMID- 11859570 TI - [What's new in pediatric urology in 1999-2000?]. PMID- 11859569 TI - [MRI diagnosis of the temporomandibular joint. Dynamic MR study of the temporomandibular joint in disk displacement: a retrospective analysis]. PMID- 11859571 TI - [Pathologic horseshoe kidney. Report of 13 cases]. AB - The authors report a series of 13 horseshoe kidneys observed over a 22-year period. All patients are men with a mean age of 39 years (20-65 years). The clinical features were dominated by abdomino-lumbar pain (nine cases). The horseshoe kidney was associated with renal stones in nine cases, uretero-pelvic junction syndrome in five cases and pyonephrosis in one case. The specific anatomic and surgical features of this rare malformation are emphasized and the therapeutic features of each uropathy associated with horseshoe kidney are discussed. PMID- 11859572 TI - [Meningeal tuberculosis and renal lithiasis. A case report]. AB - Urinary tuberculosis is frequent in Algeria. The discovery of the disease become difficult when one of the three criterium of the diagnostic does not allow a diagnosis of certitude. The authors reported the case of a 44 years-old patient admitted to hospital for tuberculous meningitis recovery from left nephrectomy for urinary lithiasis. The histology does not find specific lesions. Then, no antituberculous treatment is prescribed. The patient has developed renal and meningitis tuberculosis associated with urinary lithiasis. Koch's bacillus is found in the urine. The evolution under medical treatment was excellent. The urinary lithiasis has hided tuberculosis and the discovery of the disease was late. PMID- 11859573 TI - [Spontaneous subcapsular renal hematoma: diagnosis and treatment. Two case reports]. AB - Spontaneous perirenal haematomas essentially raise the problem of the aetiological diagnosis, but can sometimes be inadequate. Arteriography is useful when CT scan is negative or in the case of vascular disease. The therapeutic attitude, nephrectomy or conservative treatment remains controversial, but in view of the frequency of renal neoplastic lesions, the authors recommend nephrectomy at the slightest doubt concerning renal integrity. When no case be found, the assessment must be completed postoperatively with long-term, close surveillance, due to the risk of an undiagnosed neoplastic lesion. The authors report two recent cases and try to propose a diagnostic and therapeutic strategy based on the aetiology. PMID- 11859574 TI - Clear cell sarcoma of the kidney in an adult. A case report. AB - DEFINITION: Clear cell sarcoma of the kidney was initially thought to be a variant of Wilms tumour with an unfavourable prognosis. METHODS: A 23 years-old woman presented with a right abdominal mass. CT scan revealed a solid 23 cm tumour of the right kidney. A right radical nephrectomy with retroperitoneal lymph node dissection was accomplished. The histological diagnosis was clear cell sarcoma. A combination chemotherapy regimen (cisplatin and doxorubicin) was performed on six cycles. RESULTS: After two years, there was no evidence of tumour in the abdomen and thorax on CT Scan. The patient is being well. CONCLUSION: Optimal treatment is unknown, and surgery, radiotherapy and chemotherapy are used alone but mostly in combination. PMID- 11859575 TI - Follicular carcinoma of the thyroid metastasis to the kidney nine years after resection of the primary tumor. AB - The authors report a case of renal metastasis from a follicular carcinoma of the thyroid in 62 years-old man, occurring nine years after isthmolobectomy for thyroid carcinoma. Clinical symptoms radiographic results and treatment are discuss after reviewing literature. PMID- 11859576 TI - [Ureterocele in adults. Five case reports]. AB - The authors report five cases of ureterocele during six years. Mean age of the patients was 34.2 +/- 11.1 years. There were four female and one male. Pain (N = 3) and dysuria (N = 3) were the most common symptoms at examination. Diagnosis was made by ultrasound (N = 3), IVU (N = 3) or cystoscopy (N = 1). Ureteroceles were bilateral in four cases, unilateral in one case and were always intravesical with single ureter. Two patients presented lithiasis enclosed in the ureterocele. Only four patients had been operated. Surgical treatment was ureterocelectomy with ureteral reimplantation according to Cohen procedure. With mean follow-up of 10.6 +/- 4.7 months results were stable without complications. PMID- 11859577 TI - [Two rare case reports of ureteral triplication]. AB - Triplication of the upper urinary tract is rare. It is classified according to its extent and four types have been documented. Since the first description of ureteral triplication by Wrany only about a hundred cases have been reported in the literature. Paediatric cases are very few and female patients seem more concerned. The embryological event resulting in ureteral triplication derives from multiple ureteral buds arising from the Wolffian duct with fissuring of one or more of them. We report on two cases of ureteral triplication. The first one associated with an extravesical ectopic ureter and a dysplasic kidney. Diagnosis was particularly difficult and treatment consisted of total nephrectomy. The second case was associated with an upper pole hydronephrosis and a partial nephrectomy was realized. PMID- 11859578 TI - [Flow cytometric DNA analysis and cytology in diagnosis and prognosis of bladder tumors: preliminary results of a comparative study of bladder lavage]. AB - OBJECTIVE: To compare flow cytometric data (ploidy and proliferative activity or percentage SG2M-phase cells) to cytologic and histologic data of the bladder carcinomas. MATERIALS AND METHODS: Cytologic and flow cytometric analysis of DNA content were performed on 48 bladder washings: 28 bladder washings from patients being followed for urothelial carcinomas and 20 control washings from individuals undergoing cytoscopy for other reasons. RESULTS: Cytological sensitivity and specificity of bladder washing were 75% and 91% respectively. Specificity was increased to 94% using flow cytometric DNA analysis whereas sensibility was moderately decreased to 68%. Combination of flow cytometry and cytology increased the diagnostic yield to 100%. The study of the patient group showed an increased abnormalities (aneuploidy and/or proliferate activity SG2M > 10%) according to the tumor grading and tumor staging. A cytometric test was positive in 80% for G3 tumours and in 68% for G2 tumours. The staging tumor was positive in 46%, 89% and 100% of the pTa-pT1, pT2 and pT4 tumours respectively. Otherwise the comparison of control group with patients showed a statistical correlation between cytometric test, staging tumour and tumoral grading as showed in the following groups: control/G1-G2 (p < 0.05), control/G3 (p < 0.001), control/pTa-pT (p < 0.05), control/pT2-pT4 (p < 0.001). CONCLUSION: We confirmed through this study the interest of the flow cytometric DNA analysis in the diagnosis and prognosis of bladder carcinomas, and we showed the importance of the histogram classification in order to facilitate their interpretation and to avoid the trap of false aneuploidy. PMID- 11859579 TI - [Intradiverticular bladder tumors. Three case reports]. AB - The bladder tumours in vesical diverticula is rare, and the poor prognosis, because it was often with early invasion. We reported three cases of bladder tumours in vesical diverticula, with delay of diagnosis two, eight and twelve months respectively. The radiology exploration suspected the diagnosis and the histology biopsy confirmed a diagnosis of primary transitional cell carcinoma in two cases: PTa GI and T2 GII, and in an other case it was a invasive epidermoid carcinoma. The first patient was dead by urethral resection of the bladder tumour. The second required a cytoprototectomy and the last patient. The treatment consisted of radiotherapy and chemotherapy. We insisted of the particularity diagnosis, histology and therapeutic for bladder tumour in vesical diverticula and the early diagnosis in order to have a good prognosis. PMID- 11859580 TI - [Neuroendocrine tumor of the prostate. A case report]. AB - We report one case of neuroendocrine tumour of the prostate in a 74 years old patient. The clinical symptomatology was dominated by pollakuria, dysuria. Digital rectal examination showed a bulky prostate, hard and irregular. The rate of PSA was high (23 ng/mL). The endorectal ultrasonography revealed a pelvic heterogeneous mass depending of prostate. The biopsy showed an adenocarcinoma of prostate. The patient underwent a transuretral resection of the prostate (TURP) and pulpectomy. Histology and immunohistochemistry through antibody anti NES and antibody antisynaptophysin of shavings of resection revealed a neuroendocrine tumour of the prostate. Twelve months after, the lung X-ray, abdominal ultrasonography and bone scintigram was normal. The rate of PSA was 2.1 ng/mL. PMID- 11859581 TI - [Treatment of urinary incontinence in epispadias at the beginning of the third millenium]. AB - The treatment of urinary incontinence in epispadias is delicate. Urinary continence is generally obtained after onerous and repetitive surgery. Different publications show a variable results. After the study of the results of our series (nine cases) and the results published in the literature, we propose a clarification on the therapeutic modalities of urinary incontinence associated to this malformation. PMID- 11859582 TI - [When and why to use penile prostheses: seven years of experience]. AB - During the last years important changes in the management of the patients with erectile dysfunction have been verified. Psychologist, neuro-physiologist and bioengineers have given a valuable help in this field. Implant surgery represents, to our notice, the curative more acceptable choice for patient and the partner, since the results are excellent and the materials used extremely manageable. In each case the degree of satisfaction of the patient remains the only legitimate indicator currently for establish the effectiveness of the therapy. PMID- 11859583 TI - [Hemospermia: diagnosis and therapeutic aspects. Seven case reports]. AB - Hemospermia or hematospermia is a common benign condition, but its prevalence remains unknown and can result from several causes. The aetiology is idiopathic in about 30-70% of the cases. The hemospermia is first of inflammatory origin, in the young patients, where it is due to uretroprostatitis or orchi-epididymitis, but in the older, it is due to a benign of malignant prostatic tumors. Transrectal ultrasonography and magnetic resonance imaging can afforded the opportunity to best investigate the patients with hemospermia. Hemospermia is not an uncommon problem and in most instances has little clinical significance other than evoking much anxiety for the patient. In view of the literature, the authors discus the diagnostic and therapeutic approach of hemospermia through a series of seven cases. PMID- 11859585 TI - Laser treatment of congenital and acquired vascular lesions. A review. AB - Several quasi-continuous-wave and pulsed lasers can effectively treat a variety of vascular lesions. The pulsed dye laser and its newer variants were specifically designed to target hemoglobin and, by increasing their wavelengths slightly, have successfully achieved greater depths of penetration. When used in to compliance with the theory of selective photothermolysis, these systems have been shown to be safe and to have a low incidence of adverse sequelae. With the concomitant use of epidermal cooling systems, side-effect profiles have been further reduced. Although great progress has been made in the laser treatment of leg telangiectasias, even the newest laser systems have failed to meet patient expectations. Continuing advances in laser technology, however, will probably lead to enhanced clinical results, decreased side-effect profiles, improved hardware, and reduced costs. PMID- 11859584 TI - [Role of scintigraphic tests in upper urinary tract dilatation in children]. AB - The diuresis renography has an important place in the management of upper urinary tract dilatation in children. This non-aggressive and low irradiant exploration is very helpful to determine if hydronephrosis is obstructive or not and to evaluate the renal function impairment. The protocol of exam must be particularly rigorous and interpretation should take into account physiopathological factors that influence provocated diuresis. A well patient hydration and if necessary a bladder catheter are required. The MAG-3 is the radiopharmaceutical of choice especially in newborn and infants. The furosemide administration at 20 minutes after radiotracer injection is the most commonly used protocol and is generally sufficient to classify the urinary dilatation. Equivocal cases and symptomatic forms can benefit from a modified protocol in which furosemide is administered 15 minutes before radiotracer injection (F-15). False positive tests may result from a marked dilatation or renal function reduction. The place of diuresis renography in management of neonatal hydronephrosis is controversial, particularly because of the dynamic and evolving nature of this pathology. PMID- 11859586 TI - Laser resurfacing of darkly pigmented patients. AB - Hyperpigmentation following laser resurfacing in darker-skinned individuals is common but treatable. This pigmentation is usually heralded by persistent erythema and can be prevented in most patients if aggressive laser resurfacing is curtailed and conservative or single-pass techniques are applied. Avoiding excessive intraoperative frictional trauma when removing the desiccated tissue Avoiding excessive intraoperative frictional trauma when removing the desiccated tissue and potentially irritating agents or infections in the pre- or postoperative period is also instrumental in avoiding long-term complications. PMID- 11859587 TI - Regional anesthesia with monitored anesthesia care for dermatologic laser surgery. AB - This article describes an anesthetic technique that the authors have found useful for cutaneous laser surgery in keeping with the standards for office-based anesthesia practice. Although still in its infancy, office-based anesthesia for dermatologic laser procedures has become one of the most challenging yet rewarding fields of anesthesia today. As laser procedures continue to flourish, with seemingly endless technologic advances, surgeons, anesthesia providers, and other medical personnel must work collaboratively in developing appropriate office-based practice. The authors' experience using the anesthetic technique described here has demonstrated that cutaneous laser resurfacing can be conducted safely and efficiently using a combination of facial nerve blocks with intravenous sedation. PMID- 11859588 TI - Hair removal by lasers and intense pulsed light sources. AB - Laser and flash-lamp technology now offers the potential for rapid, safe, and effective treatment of unwanted hair. An ever-increasing number of published studies have confirmed the long-term efficacy of laser and flash-lamp treatment. The benefits of this technology have largely been limited to individuals with dark hair and relatively fair skin. The first studies using devices with a combination of longer wavelengths, longer pulse durations, and adequate epidermal cooling have shown that it is possible to safely and effectively treat individuals with darker skin types. The remaining challenge is to develop the means to eliminate light-colored hair as well. The increasing consumer demand for low-cost hair removal has driven the development of low-cost hair removal devices, such as small, pulsed flash lamps. The rapid pace of technologic advances and continued studies of hair follicle biology promise to improve this field over the years to come. In the future, small, low-cost laser-razors may replace all other means of hair removal. PMID- 11859589 TI - Laser treatment of epidermal and dermal lesions. AB - The CO2 laser is a versatile and effective tool for the treatment of warts and various other epidermal and dermal lesions where there is no easily targeted chromophore other than water. The development of high peak-power short-pulsed or rapidly scanned resurfacing CO2 lasers has significantly improved the safety and efficacy of this treatment modality. Many lesions amenable to CO2 laser vaporization, however, can be treated by other, far less expensive methods, and it is the laser surgeon's responsibility to use the CO2 laser only when it is demonstrably the best treatment option. The pulsed dye laser has some utility for selected recalcitrant warts. Because of their greater safety margin and more precise ablation, resurfacing CO2 and Er:YAG lasers are replacing the continuous wave CO2 laser for the ablation of many superficial lesions. PMID- 11859590 TI - Side effects and complications of laser therapy. AB - Laser skin surgery is not without risk and some degree of downtime in most situations. Patient satisfaction is key to the perceived success of the procedure. Therefore, the physician must fully explain all risks, potential complications, and expected morbidity associated with any laser treatment to be performed. Although many side effects can be avoided by use of appropriate intraoperative technique and adequate postoperative management, untoward effects may still occur and must be promptly identified and addressed. An informed patient is an important factor in the healing process and contributes to the success of the procedure. PMID- 11859591 TI - Nonpharmacologic treatments in psychodermatology. AB - The author believes that psychocutaneous medicine has indeed come of age and is being incorporated into mainstream medical practice. Patients presenting to dermatologists today are more sophisticated and are frequently dissatisfied with traditional medical therapies. They actively seek alternative approaches and adjuncts to standard treatments. In contrast to many other "alternative" (or) "holistic" treatments offered through non-medical venues, dermatologists can assure their patients that controlled studies support the efficacy of psychocutaneous techniques in improving many dermatologic conditions. Psoriasis, rosacea, herpes simplex, body dysmorphic disorder, acne, eczema, urticaria, neurotic excoriations, excoriated acne, trichotillomania, dysesthetic syndromes, and delusions parasitosis are included in this incomplete list. The author believes it is helpful for both the patient and therapist to define concrete and realistic goals for psychocutaneous intervention. Concrete observable or measurable goals can help the patient and clinician gauge therapeutic progress and success. Specifically, goals can include reduction in pruritus (rating severity from 1-10), decreased scratching activity, decreased plaque extent or thickness, decreased number of urticarial plaques, decreased flushing, decreased anxiety, decreased anger, decreased social embarrassment, decreased social withdrawal, and improved sleep. More global goals can include an improved sense of well-being, increased sense of control, and enhanced acceptance of some of the inevitable aspects of a given skin disease. Cure should never be a goal, because most disorders amenable to psychocutaneous techniques are chronic in nature; thus, cure as an endpoint would only lead to disappointment. The author encourages dermatologists to align themselves with what he euphemistically calls "a skin-emotion specialist." The skin-emotion specialist may be a psychiatrist, psychologist, social worker, biofeedback therapist, or other mental health or behavioral specialist. Patients are more likely to accept a referral to a "skin emotion specialist," because this term destigmatizes psychologic interventions. Incorporating these techniques and specialists into a clinical practice will expand therapeutic horizons and improve the quality of life of many of the patients afflicted with chronic skin disease. A final caveat must be offered about attempting to make prognostic statements regarding the likelihood of therapeutic success. Although all patients can potentially benefit from psychocutaneous interventions, those with severe psychopathology and poor pretreatment functional status are likely to be more difficult to treat and to achieve less optimal outcomes. Patients with personality disorders such as borderline, narcissistic, and schizotypal disorders, and patients with any active psychotic process certainly constitute a more resistant and difficult population with whom therapeutic success is less likely. These patients, however, are often the ones in the greatest subjective distress and certainly can profit from any of the described interventions. Quoting W. Mitchell Sams, Jr., "although the physician is a scientist and clinician, he or she is and must be something more. A doctor is a caretaker of the patient's person--a professional advisor, guiding the patient through some of life's most difficult journeys. Only the clergy share this responsibility with us." This commitment is and must always be the guiding force in the provision of comprehensive and compatient patient care. PMID- 11859592 TI - Laser surgery of leg veins. AB - The use of lasers and light sources in treating lower-extremity blood vessels has not been as successful as they are for treatment of facial telangiectasia. Several reasons for this lack of success include increased hydrostatic pressure on the lower extremities and the anatomy of lower-extremity blood vessels. These vessels are in a deeper location, have thick surrounding adventitial tissue, and have increased basal lamina when compared with facial telangiectasia. Improvements in matching pulse duration to vessel size, use of more penetrating, longer wavelengths, and the addition of sophisticated cooling methods have improved the results with leg veins during the past 2 years. Further refinements in laser technology over the next few years will probably enhance treatment efficacy of lower-extremity telangiectasias. PMID- 11859593 TI - Laser eradication of pigmented lesions and tattoos. AB - Laser treatment of pigmented lesions can be a rewarding experience when appropriate lesions are treated. Accurate diagnosis of pigmented lesions is needed before treatment. In some lesions, adjuvant topical therapy is greatly beneficial, and for others it may be the only option. The treatment of melanocytic nevi is controversial but worth pursuing. All tattoos respond well to Q-switched lasers; the appropriate wavelength depends on the color of ink. Amateur and traumatic tattoos clear readily with laser treatment. Cosmetic tattoos should be approached with caution. In addition, the use of laser responsive ink and higher-powered or shorter-pulsed (picosecond) lasers may further enhance the ability to treat tattoos. PMID- 11859594 TI - Laser scar revision. AB - A variety of lasers can be used to treat scars and striae effectively. It is of paramount importance that the type of scar be properly classified on initial examination so that the most appropriate method of treatment can be chosen. Classification also allows the laser surgeon to discuss with the patient the anticipated response to treatment. The 585-nm pulsed dye laser (PDL) is the most appropriate system for treating hypertrophic scars, keloids, erythematous scars, and striae. The PDL carries a low risk of side effects and complications when operated at appropriate treatment parameters and time intervals. Atrophic scars are best treated with ablative CO2 and Er:YAG lasers; however, proliferative keloids and hypertrophic scars should not be vaporized because of the high risk of scar recurrence or progression. The appropriate choice and use of lasers can significantly improve most scars. As research in laser-skin interaction continues, further refinements in laser technology coupled with the addition of alternate treatment procedures will allow improved clinical efficacy and predictability. PMID- 11859595 TI - Laser therapy of stretch marks. AB - Striae distensae, better known as stretch marks, are a common disfiguring skin disorder of significant cosmetic concern. Many sources have reported the use of lasers to diminish the appearance of striae. Controlled clinical studies of the various treatment modalities available for striae are relatively uncommon, and much of the clinical data are anecdotal. The use of lasers alone or in combination with other therapeutic modalities can provide a safe and effective reduction in the appearance of both red and white striae distensae. Many of these therapies require special measures for darker skin phototypes. This article reviews the historical use of laser therapy for this disorder and discusses current therapeutic options. PMID- 11859596 TI - Maximizing benefits and minimizing risk with CO2 laser resurfacing. AB - Laser resurfacing using the high-energy, pulsed and scanned CO2 laser produces the most dramatic improvement in severe photo-induced facial rhytides and deeply atrophic scars. More recently, the use of short- and long-pulsed erbium lasers in conjunction with CO2 laser resurfacing has been shown to speed healing by removing the upper layer of thermally induced necrotic tissue. The Er:YAG lasers are more superficial ablative tools that inflict less residual thermal damage on the tissue, thus allowing healing to occur without the requisite internal clearance of excessive debris. With continued research and advances in laser surgery, cutaneous resurfacing will be further enhanced with improved outcomes and lessened morbidity. PMID- 11859597 TI - Er:YAG laser skin resurfacing. AB - Recent advances in Er:YAG laser technology have dramatically enhanced the ability to resurface the skin safely. Selection of precise ablation and coagulation levels allows laser surgeons to modify these variables to fit the esthetic concerns of individual patients. These systems truly represent a viable alternative to conventional CO2 laser resurfacing. PMID- 11859598 TI - Nonablative laser skin remodeling. AB - Nonablative skin remodeling has become an attractive option for patients whose lifestyles demand a noninvasive approach to skin rejuvenation. This review addresses the rationales for therapy, specific devices, and study outcomes. PMID- 11859599 TI - What did she want with Xanax? PMID- 11859600 TI - Postmenopausal hormone replacement therapy: does the progestin make a difference? PMID- 11859601 TI - Somatization: a disorder about "nothing". PMID- 11859602 TI - Somatization: a disorder about "something". PMID- 11859604 TI - [Recovery of function and appearance--a reality question must be faced]. PMID- 11859603 TI - Factors associated with attitudes toward medication use by pregnant women, with implications for prenatal care providers. AB - Women continue to have concerns about use of a prescription medication for the treatment of infection during pregnancy. Although we surveyed adults of all ages to determine if parents of reproductive age women were harboring concerns about medication and advising their pregnant daughters to be cautious, we found it was the young women who were most concerned about the balance between medication use and impact of infection. Clinicians should be clear about the risk of infection for both the mother and the infant when medication is prescribed for infectious illness during pregnancy so women can make an informed decision about adherence to the treatment regime. PMID- 11859605 TI - [Management of deep facial burn with early postburn debridement and delayed skin grafting]. AB - OBJECTIVE: To explore optimal methods of the management of deep facial burn, so as to prevent the development of facial scar hypertrophy and deformity. METHODS: Partial tangential excision, delayed skin grafting, early relief of the eyelid eschar and immediate skin grafting were carried out in burn patients with deep facial burn during the early postburn stage. RESULTS: The wounds in 12 cases had healed completely within 3 postburn weeks(PBW). Scar hypertrophy and facial deformity were not observed during the follow-up. All the patients could express natural facial expressions. CONCLUSION: Facial deep burn wound could be managed as soon as possible with early partial tangential excision and delayed skin grafting. As a result, facial scar hypertrophy and deformity could be avoided. PMID- 11859606 TI - [Treatment of the ocular burn with hormone]. AB - OBJECTIVE: To explore new ways of the management of ocular burn with the use of hormones (human growth hormone and corticosteroid). METHODS: Twenty-one patients (30 eyes) with ocular burn were enrolled in the study. Nine cases (12 eyes) were treated with routine methods, while in 12 cases (18 eyes) corticosteroid and recombinant human growth hormone (rhGH) were used. The corticosteroid was applied immediately postburn both systemically and topically and continued for 2-4 weeks. But the rhGH was applied after 1 week postburn and continued for 2 weeks. The results in the above two groups were compared. RESULTS: Compared with those in control group, all the 18 eyes in the 12 patients with ocular burn were saved and cured with the treatments of hormones. There was significantly less incidence of corneal ulceration and perforation and better visual recovery in those treated with the hormones. CONCLUSION: Combined application of corticosteroid and rhGH might be beneficial to the management of patients with ocular burn. PMID- 11859607 TI - [Repair of the postburn scar of face, neck and chest with extensive tissue expanding technique]. AB - OBJECTIVE: To explore the treatment effects of extensive tissue expanding technique on the postburn scar of face, neck and chest. METHODS: The technique was applied in 13 patients with post-burn scar of face, neck and chest. Different number of expanders (usually 5-8) with volume of 30 ml to 450 ml each were implanted under the adjacent normal skin. The principle of plastic surgery was observed during the operation with attention to locating sear lines along leavage lines and in obscure areas. The criteria for cure were correct restoration of the position of the five organs on the face, restoration of the correct angle of jaw and neck and avoidance of skin grafting on the chest. RESULTS: All the postburn scar on face, neck and chest was removed en masse with all the flaps survived with satisfactory color and texture with this technique. In addition, the appearance and the function of facial organs and neck and chest were restored perfectly. CONCLUSION: Extensive tissue expanding technique is satisfactory in the repair of postburn scars of face, neck and chest. PMID- 11859608 TI - [Reconstruction of both eyebrows with scalp flaps with the anastomotic branch in occipital region of superficial temporal artery as the pedicle]. AB - OBJECTIVE: To explore the effects of the reconstruction of both eyebrows with scalp flaps with pedicles supplied by superficial temporal arteries. METHODS: The parietal branch of the superficial temporal and its anastomotic branch of occipital artery at temporoparietal region on the same side were designed as the pedicles, with which two scalp flaps on temporoparietal and temporo-occipital regions with reverse directions were employed to restore the eyebrows on both sides. RESULTS: Four patients were treated with this method with satisfactory appearance and survival rate (100%). The reconstructed eyebrows appeared symmetrical and heavy. CONCLUSION: Scalp flaps with the anastomotic branch at occipital region of superficial temporal artery as the pedicle might be a good choice for the reconstruction of lost eyebrows. PMID- 11859609 TI - [An experimental study of the effects of thrombin receptor activating peptide (TP508) on healing of ischemic wound and flap survival in rats]. AB - OBJECTIVE: To investigate the effects of the thrombin receptor activating peptide (TP508) on healing of ischemic wound and flap survival in rats. METHODS: Sixty six Sprague-Dawley rats were employed as the model. On the back of the rats, three kinds of wound and flap were made to establish four groups as follows: partial ischemic wound in 16, full ischemic wound in 16, routine wound in 18 and flap in 16 rats. Each group was further divided into TP508 treating group and isotonic saline control group. The total and necrotic areas of the wounds and flaps were daplicated on acetate papers and calculated with a computer on the 3rd, 7th, 10th and 14th post-operation days (PODs). RESULTS: In routine wounds, the ischemic wound area treated by TP508 was 73.7% and 45.4% in saline control groups on 7 and 14 (PODs), respectively. While in the flap model, the necrotic flap area treated by TP508 was 80.4% and 56.8% in control groups on 7 and 14 (PODs), respectively. CONCLUSION: TP508 could accelerate healing of ischemic wound and improve flap survival in rats. PMID- 11859610 TI - [A study on the myocardial contractile function and intracellular free calcium in scalded rats]. AB - OBJECTIVE: To explore the relationship between postburn myocardial function and intracellular free calcium concentration. METHODS: Sprague-Dawley (SD) rats were employed as the model in which 7 were inflicted with 43% TBSA degree scalding (S group) and 7 as sham scalding control group (C group). The hearts of the injured rats were isolated and perfused in vitro by Langendorff method after 24 postburn hours (PBHs). The hearts were connected to a cardiac function monitor, and the dynamic changes in left ventricular develop pressure (LVDP) were continuously monitored. The cardiac 19F and 31P NMR spectroscopy was determined before and after and TF-BAPTA was added to the perfusate. RESULTS: When compared with those in C group, the LVDP of scalded rat heart decreased by 40% (P < 0.01), and the myocardial cytoplasmic free calcium concentration was four times above that in C group (P < 0.01). After TF-BAPTA was taken by myocytes, cardiac LVDP only decreased by 15% to 20%, and the PCr/Pi ratio decreased, while there was no change in ATP. CONCLUSION: Myocardial contractile function could be inhibited after burn injury, which might be related to the increase of myocardial cytoplasmic free calcium concentration. PMID- 11859611 TI - [The influence of transforming growth factor beta 1 (TGF beta 1) on fibroblast proliferation and collagen synthesis]. AB - OBJECTIVE: To investigate the significance and the influence of transforming growth factor beta 1 (TGF beta 1) on fibroblast proliferation and collagen synthesis. METHODS: The fibroblasts derived from healthy human dermis and pathologic scar were harvested and cultured in vitro with TGF beta 1. The effects of TGF beta 1 on the proliferation and collagen synthesis of the fibroblasts were investigated during the early phase (24 h) of cell confluence by means of nonradioactive cellular proliferation detection, 3H thymidine and 3H proline incorporation and DNA quantification analysis. RESULTS: After the co-culture with TGF beta 1, the incorporation of 3H thymidine and 3H proline and the proliferation were enhanced significantly in the fibroblasts from pathologic scar (P < 0.01) but remained at normal level in those from healthy human dermis(P > 0.05). CONCLUSION: The cell proliferation and collagen synthesis of scar-derived fibroblasts could be positively modulated and controlled by TGF beta 1, which might be very important in the scar formation. PMID- 11859612 TI - [The influence of HOXB2 anti-sense oligodeoxynucleotides on the proliferation and expression of human umbilical vein endothelial cells]. AB - OBJECTIVE: To explore the effects of HOXB2 anti-sense oligodeoxynucleotides (asodn) on the proliferation and the expression of human umbilical vein endothelial cells (HUVECs). METHODS: Various concentrations of HOXB2 ASODN modified by thiophosphate were transfected into HUVECs by liposome mediation. MTT and RT-PCR methods were employed to determine the influence of different concentrations of ASODN on endothelial proliferation and the expression level of HOXB2 mRNA. RESULTS: After the transfection of HOXB2 ASODN, the endothelial proliferation was inhibited in dose-dependent manner. Simultaneously, the expression level of HOXB2 mRNA decreased significantly. CONCLUSION: HOXB2 might play important roles in the proliferation of endothelial cells. PMID- 11859613 TI - [Clinical application of narcotic analgesics in patients with burn and plastic surgery]. AB - OBJECTIVE: To investigate the effects of narcotic analgesics (opioids) on the patients with burn and plastic surgery. METHODS: Retrospective statistical analysis was carried out in 452 patients with burn and plastic surgery in terms of the clinical signs of pain, the number of the patients who used the drug, the kinds of the drugs used, the doses and the number of use each day. RESULTS: Inappropriate and insufficient application of narcotic analgesics was common in the past due to our incomplete knowledge of the importance of postburn and post operative pain in patients with burn and plastic surgery, and also confused concepts of the pain and misuse of opioids, often leading to insufficient analgesia. CONCLUSION: Much attention should be paid to navcotic analgesia in patients with burn plastic surgery. Some measures were suggested. PMID- 11859614 TI - [The influence of the change in postburn erythrocyte membrane viscoelasticity on the regional myocardial blood flow]. AB - OBJECTIVE: To explore the characteristics of the dynamic change in postburn erythrocyte membrane viscoelasticity and its influence on the regional myocardial blood flow. METHODS: Wistar rats inflicted with 30% TBSA III degree scalding on the back were employed as the model. The mechanical indices of erythrocytic membrane, such as elastic modulus and the coefficient of viscosity, were determined with micro-pipe sucking methods at preburn and 1, 3, 6, 12, 24 and 48 postburn hours (PBHs). The regional myocardial blood flow was simultaneously monitored with hydrogen clearance methods. RESULTS: The erythrocyte membrane viscoelasticity increased sharply at 3 PBH, reached the peak level at 6 PBH and decreased gradually thereafter to near normal level at 24 PBH. The regional myocardial blood flow decreased obviously at 1 PBH, it reached the lowest level at 6 PBH and bounced back to near normal level at 48 PBHs. The change in the viscoelasticity was significantly and negatively correlated to that of the regional myocardial blood flow. CONCLUSION: The deformability of postburn erythrocyte membrane decreased, leading to the decrease in the regional myocardial blood flow. This might be one of the important reasons of postburn myocardial injury. PMID- 11859615 TI - [An experimental study of the sciatic nerve injury by high voltage electricity in rabbits]. AB - OBJECTIVE: To explore the pathological and myoelectric-physiological changes and the mechanism of the early postburn sciatic nerve injury by high voltage electricity in rabbits. METHODS: Apparatus producing high electricity was established with a special-designed microrecorder of digital reserve. The electrical current values were determined by computer responsive analysis system with the concomitant determination of myoelectric-physiology and pathomorphology. RESULTS: When the current was set to be 50 HZ, the average indentical resistance value of the nerve tissue was (5.99 +/- 0.88) k omega and the average current value was 83.5 mA/cm2. There exhibited different degrees of sciatic nerve injury when examined by myoelectric-physiological and pathophysiological measures. CONCLUSION: Peripheral nerve could be injured by current of high voltage electricity. PMID- 11859617 TI - [Malignant low grade intestinal adenocarcinoma of the nasal cavity]. AB - Adenocarcinomata of the nose are scanty growths. They included primary adenocarcinomata not derived from salivary glands and these in its turn not share histological, pathogenical and prognostic characteristic, being the latter those of high degree or intestinal type and the other the lower degree. We report an unusual clinical case which should be classified among those of high degree, but because of its behavior showed totally benign. PMID- 11859616 TI - [The role of released cytochrome C from mitochondria in the apoptosis of HUVECs induced by hydrogen peroxide]. AB - OBJECTIVE: To explore the role of mitochondria and from which released cytochrome C in the apoptosis of HUVECs induced by H2O2. METHODS: HUVECs were cultured with H2O2 of 100 mol/L for 24 hrs. H2O2 was added after that HUVECs were precultured with CsA (cyclosporin A) for 30 mins. The cell samples were collected at different time points for DAPI staining and counting of apoptotic cell number. Simultaneously, the changes of mitochondrial permeability was observed by Rhoadmine 123 accumulation. The changes of the cytochrome C concentrations in plasma and mitochondria were determined by western blot. RESULTS: HUVECs exhibited obvious apoptosis after being processed by H2O2. The apoptotic cell number increased since 4 hrs of culturation of HUVECs with H2O2, and reached peak level at 12 hrs. And the HUVEC apoptosis induced by H2O2 could be inhibited significantly by CsA. H2O2 could lead to the decrease of mitochondrial cytochrome C concentration, which in turn lead to the increase of cytoplasmic cytochrome C concentration. Similarly, mitochondrial Rhoadmine 123 concentration decreased and the mitochondrial permeability increased. But CsA could obviously inhibit the changes of both mitochondrial Rhoadmine 123 and permeability. CONCLUSION: H2O2 could induce cytochrome C releasing to cytoplasma which lead to endothelial apoptosis. And CsA could inhibit the apoptosis by maintaining the normal function of mitochondrial membrane. PMID- 11859618 TI - [Tonsillectomy in adults. Our experience]. AB - Tosillectomy or palatine tonsils removal was by far as the very middle of the twenty century the commonest surgery in childhood, but owing to the abstinence behavior in past decades this operation has dropped considerabily and in consequence numerous grown up presents frequently tonsillar pathology requiring its removal in middle adult life. PMID- 11859619 TI - [Cryopreservation of neural allografts for the nerve reconstruction in experimental surgery]. AB - The possibility of clinical employment of nervous allografts depends on the likehood of its storage and also of the need of decreasing its antigenecity. We report our methodology for cryo-preservation and storage of nervous grafts, in view of animal experimentation as well as a re-examination of other methods in order to remember grafts storages done in previous epochs. PMID- 11859620 TI - [Deep neck infections: complications of acute pharyngitis]. AB - Deep neck infections develop as aftermath of spreading infections from an initial focus to the neighbourhood and soft tissues of the neck, through routes offering lesser resistance as fascial sheets. These cervical pictures are not numerous and generally after infective focus bad treated, linked with concomitant factors easing the rapid spread. The presence of complications as the involvement of great blood vessels, mediastin or jeopardizing the air-way may darken the prognosis, for which reason its precocious diagnosis and correct behaviour are very decisive. The paper deals with one case of neck infect, following an acute banal pharyngitis without predisposing circumstances. Follows a review of the published bibliography. PMID- 11859621 TI - [Biopsy cannula as diagnostic alternative to puncture-aspiration biopsy with fine needle in head and neck tumors]. AB - Biopsy cannula is a diagnostic tool allowing anatomopathological examination of tissue samples, achieved through a percutaneous perforation. Unlike puncture aspiration with fine needle method that only permits a cytologic study, this technique eases the removal of pieces big enough to provide the knowledge of the whole architecture of the lesion. At length in other specialities in our has been forgotten perhaps for the great efficiency of PAFN on neck masses or as complement of traditional surgery. We present one case in which the biopsy cannula showed its diagnostic usefulness. We also discuss on the technique and made a bibliographic perusal about the published literature. PMID- 11859622 TI - [Subcutaneous cervico-thoracic emphysema and pneumo-mediastinum following a tracheostomy]. AB - Reporting the case of a man, 70-year-old, suffering a chronic obstructive lung disease of moderate degree linked to important Reinke's oedema, who underwent a tracheotomy, being troubled the immediate postoperative period with subcutaneous emphysema and pneumomediastinitis, complications resolved favourably in a ten days term, and the patient could be decannulated. PMID- 11859623 TI - [Hypoglossal nerve palsy and infectious mononucleosis]. AB - Neurologic manifestations are infrequent in infectious mononucleosis and specially hypoglossal nerve involvement is very rare. To the best of our knowledge only 6 cases have been previously reported. We describe herein the case of a 20-years-old man who presented an unilateral hypoglossal nerve palsy secondary to infectious mononucleosis. He was treated i.v. steroids without recovery, but the palsy resolved spontaneously 2 months after the onset. PMID- 11859624 TI - [Head and neck abscesses]. AB - OBJECTIVE: To review the clinical, microbiological and therapeutic characteristics of all head and neck abscess seen in a Community Hospital. MATERIALS AND METHODS: Medical records of all patients admitted to Sierrallana Hospital (Torrelavega, Cantabria) from January 1995 till June 1998, because of head and neck abscess were reviewed. RESULTS: Fyfty-seven head and neck abscess of 54 patients were considered. Age of patients ranged from 14 to 78-year-old. Periamygdaline region was the most frequent location, where as tonsillar and dental infections were the origins of the abscess in the large majority of cases. Most of the abscess cultures yield mixed flora. Intravenous penicillin was used in 77.3 percent of patients and surgical drainage was required in almost 80 percent of cases. Treatment response was good with no serious complications in any case. CONCLUSIONS: Head and neck abscess are a relative common cause of hospital admissions. Among them periamygdaline abscess are frequently found. The treatment is surgical drainage with local or general anesthesia (depending of location) and simultaneous i.v. course of antibiotic (Penicillin G sodium salt) efficacious in the great number of patients. Serious complications are very rare. PMID- 11859625 TI - [Granular cell tumor. A lingual case report]. AB - Although the origin of granular cell tumor (GCT) is no totally clarified, a neurogenic source is generally accepted. Some think its forerunners are the Schwann cells. Despite the tongue's localisation is frequently seen, few are the papers dealing with this topic. We report the case of a patient suffering a granular cell tumor of the tongue and profit the opportunity to reconsider the followings items: clinical findings, examination, diagnosis, histology and management and course of these growths. Literature's review of the theme. PMID- 11859626 TI - [Metastatic carcinoma of the parotid gland. Review of the literature. Report of two cases]. AB - The great majority of parotid gland tumours are benign. Metastatic malignant ones are scarce in number. The source of these metastasis are growths localized either in head and neck or distant tumours spreading on intra- or periparotid lymph nodes. Two cases of metastatic parotid growths diagnosed and treated last year are reported. Bibliographic review of the topic. PMID- 11859627 TI - [50 years professional practice in child psychology and child psychiatry]. PMID- 11859628 TI - [Psychoanalytically based psychotherapy]. AB - Depth psychologically founded psychotherapy is a treatment based an psychoanalysis. This article describes the treatment method. The following aspects are especially emphasized: Terminology definitions, historical development, specific differences between depth psychologically founded and analytic psychotherapy, and the treatment practice with the focus an current psychosocial conflicts and the implications connected with these. Then indication, counter-indications and differential indication are discussed, and the status of empirical evaluation is described. Finally the therapist's qualification requirements necessary for this treatment method are explained. PMID- 11859629 TI - [Psychoanalytically based psychotherapy with children and adolescents]. PMID- 11859630 TI - [Psychoanalytically based psychotherapy with children and adolescents]. AB - The author discussed the term "depth psychologically founded psychotherapy" for children and adolescents in its current meaning within analytic psychotherapy for children and adolescents, and also the possible changes due to the Act for Psychotherapists ("Psychotherapeutengesetz"). A differentiated description what depth psychologically founded psychotherapy comprises is based an a paper which the author worked out together with a group of psychotherapists for children and adolescents. The term "analytic oriented psychotherapy" is brought up for discussion which the author views as being more adequate for the presented form of psychotherapy. The peculiarities of the technique used in depth psychologically founded psychotherapy with children, adolescents and persons the children/adolescents relate to are described in three examples. PMID- 11859631 TI - [Psychoanalytically based and psychoanalysis in children and adolescents from the viewpoint of a child and adolescent psychotherapist]. AB - The author, practitioner of psychoanalytic psychotherapy with children and adolescents, explains her point of view regarding psychotherapy versus psychoanalysis. Verbatim process notes illustrate the work and are used to plan the therapeutic method. PMID- 11859632 TI - [Psychoanalytically based psychotherapy in childhood and adolescents from the viewpoint of expert assessment]. AB - Depth psychologically founded or analytically psychotherapy with children and adolescents is not only a question of the respective "Fachkunde", a variable referring to the therapist and his training. Aspects of differential indication and effective factors pertaining to psychoanalytic techniques must also be taken into consideration. There are different techniques which should be conceptulized as a continuum. The effective factors vary in their impact. It seems problematic to distinguish between single discrete techniques. Depth psychologically founded psychotherapy with children and adolescents is a technique characterized by a limited goal which can be formulated in a "focus". Special emphasis has to be laid on the environment and the present state of development. First findings from a workshop on focal psychotherapy with children and adolescents at the Psychoanalytischen Institut Bremen, are discussed by case illustration. PMID- 11859633 TI - [Thirty years later....]. PMID- 11859634 TI - The ongoing assessment of the supply of medical doctors in Denmark. PMID- 11859635 TI - Methods for assessment of future demand of medical doctors in Denmark. AB - BACKGROUND: Whereas supply prognoses is mainly a matter of establishing mathematical scenarios with ongoing adjustments of variables, reliable demand prognoses are more difficult and complex to establish. However, optimum use of human and educational resources and--on the other hand--sufficient supply of medical doctors calls for reliable supply as well as demand prognoses. METHODS: Based on a number of technical and political considerations the first demand prognosis was based on three different methods; 1. Changes in demand due to demographic changes. 2. An evaluation made by the owners of the hospitals. 3. An evaluation made by the scientific medical societies. RESULTS: The demand prognosis was found to be hampered by a number of limitations. Demographic changes beyond the next 10 years was found to be unreliable. Moreover, the demand observed during the last 20 years was only to a very limited extent explained by demographic changes. The owners of the hospitals could only prognosticate for an election period, i.e. for about 5 years,--and only in very general terms that were difficult to interpretate. From some counties the answers did only seem to be based on rather superficial analysis and gave very little information. Each of the Scientific Medical Societies claimed increasing demands for the next 25 years, despite marked expectations for diagnostic and therapeutic advantageous technological developments. The overall conclusion from the combined weighed analysis was an expected increase in the demand of medical doctors of about 1% per year in the short as well as in the long term (25 years). This should be compared to the observed development during the last 10 years, i.e. an annual increase of 1.7-1.8%. PMID- 11859637 TI - A model for the long-term planning of physician workforce in Hungary. AB - A computer programme for the long term (25 years) projection of the physician workforce was developed. The input variables were the admittance rate of the medical schools and the unemployment rate of the active physicians, the output variables were the number of active physicians. The age and sex structure, the retirement age were taken into account. Different scenarios were made to compare the impact of the output of medical faculties and the unemployment rate of doctors on the number of active of the physicians in five years intervals for a twenty-five years period. The input of the young graduates varied between 800 600/year, the unemployment rate between 4.5% and nil. At end of the 25 years projection period the reduction of physicians was between 9.6 and 25.0%. The aim was to find a professionally acceptable, socially tolerable and politically eligible optimal relationship among the three variables. PMID- 11859636 TI - Dimensioning of the postgraduate educational training in Denmark. AB - BACKGROUND: Postgraduate medical training in Denmark consists of basic training, offered to all medical doctors, followed by specialist training. The National Board of Health is responsible for the overall frame of medical training in Denmark and determines the number of trainee positions for each of the 42 specialities available. The total number of positions and their distribution between specialities are based on demand, supply and demographic considerations. Approximately 85% of medical doctors finalise specialist training. METHODS: Denmark is divided into three educational regions: North, South and East. Each region consists of counties, with their own administration. The National Board of Health approves each position for postgraduate training. All new positions and changes of existing positions are evaluated before approval by The National Board of Health. An updated version of the list of educational positions is available on the internet. CONCLUSION: The Danish governmental authorities have an efficient tool to control the dimensioning of the postgraduate medical education and thereby the production of specialists. Medical doctors can easily get information about where to obtain trainee positions. The majority of hospital departments improve their recruitment potential by participating in medical training. A number of structural changes, for instance establishing of medical centres and corporations within larger entities across an extended geographic area, specialisation between hospitals and reduced number of hospitals, calls for decentralisation of the administration in order to improve flexibility in the organisation of postgraduate medical training. However, the National Board of Health will still co-ordinate at the national level. PMID- 11859638 TI - Health human resource reform in Tajikistan: part of a masterplan for change. AB - Like many countries of the former Soviet Union, the Republic of Tajikistan inherited a poorly paid physician workforce dominated by specialists. This Central Asian republic has been forced to move slowly to change the physician workforce and to implement primary health care. Several years of civil war following independence in 1991 made reform of the struggling health system politically and economically difficult. The civil war also resulted in a loss of health personnel, with significant numbers of physicians leaving the country. The low pay of health professionals caused others to move to higher paying jobs in non-health related professions. A comprehensive masterplan for the reform of the health care system that has been developed through a participatory process is in the process of formal approval. The human resources component of the health care reform masterplan calls for a shift to emphasize the role of primary health care and the introduction of family physicians (FPs) as the cornerstone of the primary health care. With only 90 family practice physicians trained in 2000, the country faces a massive task in retraining existing physicians and training new FPs. The first 40 medical students to enter training as FPs are scheduled for 2001. Retraining at the Post Graduate Institute will be supplemented in 2002 by programs in the three oblasts. To overcome the shortage of FPs a comprehensive job analysis and workload assessment will be conducted to redefine the role of health professionals and involve others in the provision of care. Historically nurses have not been allowed to perform to their full capability and physicians have performed tasks more suitable for mid-level personnel. A strategy to solve maldistribution problems and to develop incentives to stem the loss of physicians will be also implemented. While circumstances have forced the Republic of Tajikistan to move slower than other countries to reform the inefficient health system inherited from the Soviet Union, the current masterplan for the entire system shows a commitment to change. The Masterplan, along with lessons from what has occurred in other countries, provide an opportunity for a well-ordered reform of health human resources within the overall context of health system reform. PMID- 11859639 TI - Physician workforce reform in Lithuania: an inevitable transition. AB - The health system in Lithuania before independence was the same centralized system, the Semashko model that was prevalent throughout the former Soviet Union. The Soviet system focused more on quantity than quality, which resulted in a greater number of physicians per 100,000 than found in Western Europe. As in other Central and Eastern European countries, the health system in Lithuania has undergone substantial change since 1990. A major part of the reform of physician workforce in the former communist countries has been development of primary health care as a centerpiece of the health systems. This move in Lithuania has been hindered to some degree by the shortage of general or family practitioners. The focus on specialists in the old Soviet system led to a severe shortage of physicians in general practice. Only about 10% of physicians were general practitioners (GPs) in 1993 compared to a European Union average of 35%. That percentage had increased to 16% by 1998. Programs have been established to both train new GPs and to retrain specialists to become GPs, although a shortage of teachers and finances are problems facing these programs. Although the physician workforce in Lithuania has undergone substantial reform since 1990, real change has been slow. The physician to population ratio in Lithuania has failed to decline, moving from 387 per 100,000 population in 1991 to 398 in 1997, before dropping to 395 in 1998. The ratio remains substantially higher than current EU averages, reflecting a difficulty in reducing the number of physicians. Here we review what has happened to the physician workforce in Lithuania since independence and evaluate policies implemented. Efforts to reduce the number of physicians are also assessed. The results in Lithuania are compared to those in other CEE countries and to EU averages. PMID- 11859641 TI - Specialization of doctors, general practice and the training system. PMID- 11859640 TI - Access to health services, geographic distribution of medical doctors and their orientating programs. PMID- 11859642 TI - Local health care markets and provider mobility. PMID- 11859643 TI - Doctor substitutes and doctor extenders: the other medical workforce. PMID- 11859644 TI - Intracranial complications of sinusitis. AB - OBJECTIVE: The aim of this study was to discuss the presenting signs and symptoms important for the early diagnosis and to review the medical and surgical management of patients presenting with intracranial complications of sinusitis. METHODS: Retrospective review of the medical records of all patients admitted with intracranial infections between 1990 and 1999 at the Department of Otolaryngology, Department of Neurosurgery and Department of Infectious Diseases, University of Cluj, Romania. RESULTS: Sixteen patients had 23 intracranial complications of sinusitis. These were meningitis (6 cases), frontal lobe abscess (6 cases), epidural abscess (5 cases), subdural abscess (4 cases), and cavernous sinus thrombophlebitis (2 cases). In almost all patients, early symptoms included headache and fever. Clinical diagnosis was confirmed by imaging. Management of these infections included intravenous broad-spectrum antibiotics, appropriate neurosurgical and sinus drainage. Among these patients, there was one death and 4 patients developed significant neurologic sequelae. Poor prognosis was associated with the presence of subdural abscess and a prolonged time interval between the onset of symptoms and the beginning of treatment. CONCLUSION: Early diagnosis of intracranial complication of sinusitis can reduce morbidity and mortality associated with these complications. PMID- 11859645 TI - Tracheoesophageal fistula: case report and review of literature. AB - The case of a 44 year old woman with progressive postoperative dysphagia and food inhalations complicated by recurrent pulmonary infections, due to a tracheoesophageal fistula (TEF) is reported. Some months earlier, this woman had been operated for a cerebral aneurysm with hemiplegia and aphasia. For several months, pulmonary and feeding difficulties had been attributed to neurological status. Wide TEF was diagnosed by bronchoscopy, confirmed with fistulography. Surgical closure was performed: the oesophagus was sutured, and covered with fascia and a segment of the cervical trachea was resected with end-to-end anastomosis. Acquired nonmalignant TEF is an uncommon disorder with a high degree of morbidity and mortality. The etiology of those TEF is still unclear: traumatic intubation, elevated endotracheal tube cuff pressure, nasogastric tube, inflammation, poor general conditions,.... A better knowledge of the predisposing factors and physiopathology could decrease the number of acquired TEF. PMID- 11859646 TI - Spontaneous onset of CSF otorrhea from a facial canal fistula in an adult: case report. AB - The purpose of this report is to describe a 64-year-old woman who presented an 8 month history of recurrent spontaneous cerebrospinal fluid leakage into the left middle ear. High resolution computed tomography and computed tomography cisternography showed only a mild protrusion of the tympanic segment of the left facial canal into the ipsilateral middle ear but no definitive fistula. At surgical exploration, cerebrospinal fluid leakage resulted from a fistula in the tympanic segment of the left facial canal. In conclusion, in the adult patient reported herein, the fistula leading to spontaneous CSF otorrhea was identified definitely only intraoperatively. Furthermore, although there were no signs or symptoms of facial nerve dysfunction, it was located in a rare site such as the second segment of the facial canal. When the clinical history, physical examination, and laboratory analysis are strongly suggestive for spontaneous CSF otorrhea, surgical exploration is mandatory even if neuroimaging evaluation is negative or questionable. PMID- 11859647 TI - Bilateral chylothorax after left radical neck dissection. AB - Bilateral chylothorax, as a complication of neck dissection, is very rare as evidenced by the 11 cases reported in the literature up to date. We present an additional case of bilateral chylothorax following a left radical neck dissection and concomitant chylous neck fistula. This case was successfully treated by chest drainage and total parenteral nutrition. Early diagnosis of chylothorax is urged due to the consequences on metabolism and respiratory conditions. Management by aspiration drainage is usually sufficient to control pleural effusions. PMID- 11859648 TI - Fibromyxoma of the middle ear (a case report). AB - Fibromyxoma of the middle ear (a case report). We report an extremely rare case of fibromyxoma of the middle ear which was first suspected to be a cholesteatoma. Surgery was performed through a postauricular incision and pathologic study of the tumor gave us the final diagnosis. Fibromyxoma is an uncommon benign tumour of the head and neck region which usually arises in the jaw (1). Its local aggressivity and gelatinous consistency explain the difficulties to remove it radically and its frequent recurrences after treatment (2). Surgical resection with a wide excision seems to be the treatment of choice. Some authors advocate the use of Argon or CO2 laser to achieve the treatment. A review of the poor literature about this subject is discussed in this article. PMID- 11859649 TI - Ectopic cervical thymus: case report and review of the literature. AB - The authors present a case of ectopic cervical thymus in a 4-month-old child. Ectopic cervical thymus is an uncommon etiology of a neck mass in a child. It is often misdiagnosed preoperatively as a possible malignancy. It is usually located along the embryologic course of thymus migration from the angle of the mandible to the superior mediastinum. Its treatment is surgical since malignant transformation and respiratory infection have been described. No case of immune deficit after total thymectomy has been described in the literature. PMID- 11859650 TI - Is septal deviation a risk factor for chronic sinusitis? Review of literature. AB - INTRODUCTION: The purpose of this work is to examine the role of septal deviation in adults in the pathogenesis of chronic sinusitis. This evaluation would allow a better understanding of the contributing factors to this pathology and would improve results in their management. MATERIAL AND METHODS: This review was based on a Medline research over English and French literature published between 1980 and May 2001. Twenty-five papers were scrutinized. RESULTS: Few investigators examined the role of the deviated septum on sinus disease. Methodologies and results are often contradictory. Furthermore, these studies are mainly based on radiological imaging. In most studies, prevalence of septal deviation looks the same when having compared among patients with radiological rhinosinusitis and among general population. An increased incidence and severity of sinus disease correlated to an increasing angle of septal deviation in the ostiomeatal complex area is reported by Calhoun (7), Youssem (4), Matschke (18) et Elahi (20). One study (1) demonstrates a significant association between the shape of the nasal septum and the location of the sinus disease: septal crest is associated with homolateral sinusitis, "watch-glass shaped diformity" with controlateral sinusitis. Stammberger (21, 25) suggests a pathophysiological role of the septum through a mechanical obstruction on the ostiomeatal complex; Danese (1) and Blaugrund (19) by an alteration of the ciliary activity secondary to a modified air flow. Bachert (24) demonstrates a connection between septal deviation and antral ventilation. CONCLUSION: This literature review cannot establish a definite role to the nasal septum neither as the pathogenesis of chronic sinusitis nor as a contributing factor. No relationship between septal surgery combined with sinus surgery, and the postoperative prognosis nor on the subjective comfort of patients can be demonstrated. Subsequently, it appears that performing septoplasty only aims at relieving nasal obstruction complaint or at improving surgical access to the ethmoid sinus. PMID- 11859651 TI - Efficacy and safety of levocetirizine in seasonal allergic rhinitis. AB - OBJECTIVE: Pharmacodynamic studies have demonstrated that levocetirizine is the active enantiomer of cetirizine. This first therapeutic trial of levocetirizine aimed at determining the dosage with the best benefit/risk ratio in patients with seasonal allergic rhinitis (SAR). METHODS: Patients with seasonal allergic rhinitis were randomised in a placebo-controlled, double-blind, parallel-group multicentre study 2.5, 5, 10 mg levocetirizine or placebo once daily during 2 weeks. Patients filled in a diary evaluation card every evening before taking study medication using the classical (0-3) scale for assessment of severity of sneezing, rhinorrhea, nasal congestion, nasal pruritus and ocular pruritus over the preceding 24 hours. The Total Four-Symptom Score (T4SS) was calculated by adding the individual symptom scores, excluding nasal congestion. RESULTS: 470 patients were included and constituted the intent-to-treat population. All 3 doses of levocetirizine were significantly superior to placebo in reducing the mean T4SS over the 2 weeks (all P (0.001). Additionally, individual symptom severity scores for sneezing, rhinorrhea, itchy nose, and itchy eyes were also significantly decreased for all doses of levocetirizine. Levocetirizine was significantly superior to placebo in reducing symptom severity with an important global treatment effect (P = 0.0001), except for nasal congestion. Furthermore, there was simple linear relationship between levocetirizine dosages and reduction of T4SS (P = 0.001). All doses were well tolerated, somnolence was higher with 10 mg (10.2%) than 5 mg (1.7%) and other adverse events were more frequent with the highest dose. CONCLUSION: Levocetirizine 5 mg once daily has an optimal benefit/risk ratio in the treatment of SAR. PMID- 11859652 TI - [Competitive morbidity ant its impact on life expectancy: evaluation and inclusion in the therapeutic decision regarding localized prostatic cancer]. AB - OBJECTIVES: The treatment decision taken by a multidisciplinary meeting for patients with localized prostate cancer must take into account the clinical stage of the cancer and its histological characteristics, but also the patient's age, general state and any concomitant diseases, as treatment is only beneficial when it induces a reduction of morbidity and specific mortality. The specific survival with or without recurrence after treatment for localized prostate cancer is long, at least more than 10 years. Curative treatment is generally not proposed to men with localized prostate cancer when his probability of survival related to a competitive morbidity (intercurrent medical disease) is estimated to be less than 10 years. The objective of this study was to measure the increase or reduction of the survival probability of a patient with localized prostate cancer according to his competitive morbidity, based on the mean life expectancy of the general population. METHODS: Review of the literature. RESULTS: Studies describing the natural history of prostate cancer show that the impact of treatment on morbidity of the cancer (local and/or metastatic) requires a life expectancy of about 8 to 10 years. The impact of a treatment on specific survival requires a life expectancy of about 13 to 15 years. The exact prevalence of diseases coexisting with prostate cancer is unknown. In the USA, The Index of Coexisting Disease (ICD), which takes into account 14 diseases, appears to be the most reliable tool to measure the competitive morbidity in patients with localized prostate cancer. Each disease is classified into 4 levels of severity (score 0 to 3). A table indicates estimated life expectancies by age-group and by ICD score. All men with a high score (2 to 3) die within 10 years after diagnosis, men with a score of 0 have a better estimated life expectancy according to age than that of the general population. CONCLUSION: The upper age limit, theoretically set at 70 years, in order to propose curative treatment for localized prostate cancer needs to be reviewed (the mean life expectancy for a 70-year-old man is 12.9 years in France). According to the ICD, the life expectancy at 70 years is 14.8 years in the case of a score of 0 and 8.4 years in the case of a score of 2. In the case of a score of 2, the impact of curative treatment on localized prostate cancer would be real on morbidity, but not on specific mortality. PMID- 11859653 TI - [Biphosphonates in the treatment of bone metastasis of prostatic cancer]. AB - Bone metastases are common in patients with prostate carcinoma and they are the most frequent cause of skeletal morbidity. Most of these metastases ar osteocloritic, but it has been shown that the abnormal osteoblastic bone formation is preceded by osteoclastic activity which appears to be associated with bone pain providing the rationale for using biphosphonates as palliative therapy. In a recent study, soledronate has been shown to be effective in patients with osteoblastic bone destruction. This is the first time any bisphosphonate reduces skeletal morbidity. PMID- 11859654 TI - [Evaluation of clinical reasoning in urology: contribution of the Script Concordance Test]. AB - OBJECTIVE: The Script Concordance test is designed to measure knowledge organization, to allow interpretation of data in clinical decision making. The objective of this study was to assess the value of this new written evaluation instrument to evaluate clinical reasoning in urology. MATERIAL AND METHOD: An 80 item examination was completed by a group of medical students (n = 15), a group of urology interns (n = 11), a group of registrars-assistants (n = 7), and a group of experienced urologists (n = 10). The scores obtained were compared by analysis of variance. The reliability of the test was studied by calculating Cronbach's coefficient alpha. RESULTS: The mean score was 46.95 +/- 6.80 for students, 56.18 +/- 1.73 for interns, 66.27 +/- 4.92 for registrars and 63.38 +/- 2.19 for urologists. The differences observed between the scores for students, interns and urologists were significant. The reliability coefficient was 0.79 for the entire examination. CONCLUSIONS: This test is able to discriminate various levels of experience in urology. It proposes a simple and direct approach to evaluation of knowledge organization. Further studies are necessary to confirm the contribution of this test to the strategy of evaluation of the clinical skills in urology. PMID- 11859655 TI - [Esthetic and functional results of the transperitoneal laparoscopic approach in nephrectomy]. AB - OBJECTIVE: To evaluate the aesthetic and functional sequelae of laparoscopic transperitoneal nephrectomy. MATERIAL AND METHODS: Twenty-one laparoscopic transperitoneal nephrectomies were performed between 1996 and 1999. Four trocars were used in 9 patients and 3 trocars were used in 12 patients. In all patients of the series, nephrectomy was performed without manual assistance and the kidney was extracted from the iliac fossa after enlarging a trocar orifice. A questionnaire evaluating the aesthetic sequelae of the operation, resumption of everyday activities and the patient's general satisfaction was sent to each patient by mail. RESULTS: 17 patients completed the questionnaire after a mean follow-up of 12.2 months (range: 2 to 33 months). Scars were cosmetically satisfactory in 100% of cases, painless in 100% of cases and were considered to be invisible in 58.8% of cases. All patients were satisfied with the operation, but only 70.6% would have recommended this procedure to a friend or relative. 57.1% of the patients hospitalised for less than 5 days considered the hospital stay to be too brief and 42.9% considered it to be barely sufficient. Time to resumption of everyday activity varied considerably (7 to 70 days) with a mean of 32 days and was not correlated with operative complications. CONCLUSION: Laparoscopic transperitoneal nephrectomy achieves good aesthetic and functional results, but patients are not satisfied with the short hospital stay. Resumption of everyday activity does not appear to depend on the postoperative course. PMID- 11859656 TI - [Treatment of uretero-intestinal and uretero-vesical stenoses with the Acucise balloon catheter]. AB - OBJECTIVE: Acucise balloon catheter has been proposed as an alternative to open surgery for the treatment of strictures of the ureteropelvic junction because of its low morbidity and the short hospital stay following the endoscopic procedure. The objective of this study was to evaluate the results of this technique applied to patients developing strictures after surgical reimplantation of the ureterovesical (UV) or uretero-intestinal (UI) junction. MATERIAL AND METHODS: Between March 1997 and January 2000, 12 strictures (11 patients) were treated by Acucise balloon catheter via an antegrade and/or retrograde approach with double J stenting for an average of 6 weeks (range: 4 to 12 weeks): 6 uretero-ileal strictures (3 Bricker, 1 uretero-ileoplasty, 1 enterocystoplasty and 1 Kock pouch) and 6 ureterovesical strictures (Lich-Gregoir or Faquin UV reimplantations after gynaecological, vascular or endoscopic surgery). The median postoperative follow-up was 16 months (range: 10 months-36 months). A good result was defined by the absence of recurrence of the stricture evaluated both clinically and radiologically (regression of stasis measured by IVU and/or ultrasonography). RESULTS: The mean operating time was 70 min and the mean hospital stay was 4.8 days (range: 3 and 14 days). Only one intraoperative complication was observed (migration of the double J stent to the kidney). The operation was successful in 8 patients (75%). The success rate was 83% for ureterovesical strictures and 50% for uretero-ileal strictures. A history of previous irradiation appeared to be a factor of failure. CONCLUSION: The Acucise procedure is a minimally invasive and effective (75% success rate) treatment option for the treatment of postoperative stricture after ureteric reimplantations. In our department, this option is considered to be first-line treatment, as surgical reimplantation is reserved for failures of the endoscopic technique. PMID- 11859657 TI - [Tumors of the upper urinary tract: results of conservative surgery]. AB - OBJECTIVE: To determine the results of conservative surgery for upper urinary tract urothelial tumours. PATIENTS AND METHODS: From 1986 to 1997, 352 patients were treated in the Belgrade urology clinic for upper urinary tract urothelial tumour. 54 patients (15.3%) were treated by conservative surgery. The sex ratio was 1.3 men for 1 woman. The mean age was 63 years. In most cases, the tumour was situated in the ureter. Conservative surgery was performed on principle in 60% of patients for a small isolated lesion (solitary low-stage, low-grade tumour). In contrast, in about 40% of cases, conservative surgery was performed by necessity due to the presence of bilateral tumours, a solitary kidney or renal failure related to Balkan nephropathy. The median follow-up was 67.3 months (range: 6 months-14 years). RESULTS: 15.8% of patients developed a local recurrence during the follow-up period. The risk of recurrence was higher when conservative surgery was performed for indications of necessity than when it was performed on principle (21.7% versus 11.8%), but the difference was not statistically significant (c2 test, t test). The stage and grade of differentiation were identified as the most significant predictive factors for the risk of local recurrence. The overall 5-year survival rate was 67% with more favourable results in the case of conservative surgery performed on principle compared conservative surgery by necessity (72% versus 60%). The difference between these results was not statistically significant, but a statistically significant difference was observed for tumour stage and grade (grade III versus grade I and II, pT3 versus pT1, pT2). The 5-year survival probability was 68.5%. Recurrence was most likely to occur during the early postoperative course, as 81.56% occurred during the first 18 months. CONCLUSION: Urothelial tumours can be managed conservatively. However, the risk of recurrence is directly correlated with the tumour stage and grade, with a high level of statistical significance, and with the type of indication for conservative surgery performed, but with no statistically significant difference. PMID- 11859658 TI - [Role of antibiotic prophylaxis in ambulatory cystoscopy]. AB - BACKGROUND: There has been a great deal of discussion regarding the necessity of antibiotic prophylaxis in transurethral cystoscopy. In order to clarify this complicated issue, a randomized prospective study was performed on 126 patients planned for cystoscopy. PATIENTS AND METHODS: 126 patients who underwent urethrocystoscopy and did not have pyuria and bacteriuria were included and divided randomly into 2 groups: group 1 received 400 mg of norfloxacine and group 2 nothing. Urinalysis were performed on all patients 3 days after the examination. Statistical analyses were performed using Chi 2 test and the level of significance was set at 5%. RESULTS: The global rate of infection was 5%. In the group 1 the incidence of infection was 3% (2/67) vs 5.1% (5/59) in group 2. There were no significant differences in the incidence in the background factor between the 2 groups of patients (p > 0.05). CONCLUSION: Prophylactic administration of antibiotic before cystoscopy does not decrease the incidence of urinary tract incidence and it is not necessarily in patients with sterile urine. PMID- 11859660 TI - [Prevalence of nocturia among people in Auxerre: the French part of the UrEpik study]. AB - OBJECTIVES: To estimate the prevalence of nocturia among men and women aged 40 to 79 years old. MATERIAL AND METHODS: A postal survey was carried out in a community-dwelling random sample in Auxerre, France. Nocturia was assessed using the item 7 of the International Prostate Symptom Score (IPSS). RESULTS: 1216 men and 591 women participated in the survey. Average nocturnal urinary frequency was 0.98 and 0.95 among men and women, respectively. Between the age ranges 40-49 years and 70-79 years, the frequency increased from 0.70 to 1.61 among men, and from 0.72 to 1.31 among women. In the age range 70-79 years, 48.1% of men and 31% of women awakened from sleep to urinate at least twice per night. CONCLUSION: Patients and primary care physicians should be more informed about screening and management options, and impact of nocturia on quality of life and related morbidity. PMID- 11859659 TI - [Urinary leukocytes as a new prognostic marker of therapeutic response and of adverse effects associated with the maintenance treatment with endovesical BCG, for the prophylaxis of superficial bladder tumors]. AB - The use of Bacillus Calmette-Guerin (BCG) vaccine as immunotherapy for superficial bladder cancer constitutes a major progress, although it remains limited by the development of adverse effects and problems related to safety. No individual tool is currently available in clinical practice to predict the efficacy or adverse effects of BCG therapy. These problems are accentuated in the case of maintenance therapy according to the protocol of D.L. Lamm. OBJECTIVES: To define whether the urinary leukocyte count observed after intravesical BCG instillation could be associated with adverse effects and efficacy of treatment. MATERIAL AND METHODS: A cohort of 72 consecutive patients (518 instillations) was studied prospectively. We defined four classes of adverse effects associated with BCG, according to their type, severity and duration. In combination with this classification, we performed systematic optical urinary leukocyte count on the third day after each instillation (KOVA-Slide 10). RESULTS: A high urinary leukocyte count (cut-off value: 165,000/ml) was correlated with absence of recurrence (p = 0.009). The adverse effect classification also demonstrated that urinary leukocyte count was related to the severity and duration of adverse effects (p < 0.0001). Median values observed for class I, II and III adverse effects were 40,000 leukocytes/ml, 150,000 leukocytes/ml and 350,000 leukocytes/ml, respectively. No class IV adverse effects were observed. The risk of developing class III adverse effects was increased when the urinary leukocyte count was greater than 86,000 leukocytes per ml. CONCLUSION: These results suggest a probable relationship between efficacy and safety of BCG, during maintenance therapy. Randomized prospective studies are necessary to evaluate urinary leukocyte count as a tool for adaptation and optimization of BCG therapy. PMID- 11859661 TI - [Can a precise vesiculectomy be performed during radical prostatectomy?]. AB - OBJECTIVE: To define the anatomical landmarks allowing precise vesiculectomy to be performed during radical prostatectomy for cancer. MATERIAL AND METHODS: 12 non-formalin-preserved anatomical subjects were dissected as during retropubic and perineal prostatectomy, with and without coloured latex vascular injection. RESULTS: Three anatomical landmarks were defined: 1) Denonvilliers' fascia; 2) vas deferens; 3) arteries supplying the seminal vesicles. CONCLUSIONS: These three anatomical landmarks ensure: complete resection, preservation of adjacent anatomical structures and elective haemostasis. PMID- 11859662 TI - [Pharmacologic stimulation of ejaculation with midodrine hydrochloride (Gutron) for medically assisted reproduction in spinal injury]. AB - INTRODUCTION: Midodrine hydrochloride (Gutron) is proposed to induce ejaculation in spinal injury patients desiring paternity as an alternative to vibromassage, electrostimulated ejaculation and surgical collection of spermatozoa. The authors report their experience in 10 spinal injury patients. PATIENTS AND METHODS: 14 trials of pharmacologically-induced ejaculation were performed in a context of medically assisted reproduction (MAR) in 10 spinal injury patients (7 with paraplegia > T11; 1 with paraplegia < or = T11; 2 with quadriplegia) an average of 4.5 years after the injury. Patients had a mean age of 28.5 years (range: 18 to 36 years). Nine had persistent reflex erections. After IC injection of prostglandin E, 10 to 30 mg of Gutron was administered by slow i.v. infusion. Spermatozoa were collected during antegrade ejaculation and/or in previously alkalinised urine. RESULT: Ejaculation was obtained in 10 cases (71.4%), either antegrade (7 cases), or retrograde (3 cases). The 4 failures corresponded to ejaculation failure in 3 cases and adverse effects of Gutron (hypertension) in 1 case. However, storage of spermatozoa could be performed in only 4 cases (40%), as pyospermia or severe necrospermia were observed in 6 cases. CONCLUSIONS: Midodrine hydrochloride gives good results in terms of ejaculation in spinal injury patients. However, the quality of semen collected is often poor due to the long interval since the initial trauma. Midodrine hydrochloride, ideally used after antibiotic treatment, can nevertheless constitute an alternative to other techniques. PMID- 11859663 TI - [Kidney cancer and pregnancy]. AB - Renal cancer is rare in pregnant women. Through a study of two cases, the authors underline diagnostic, therapeutic and pathogenic aspects of this association. Two female patients, 26 and 34 years old, were admitted for renal tumor, discovered respectively at the first and the second trimester of pregnancy. Investigation's exams for tumor extension were negative. They were based on ultrasound exam (2 cases), CT scan (1 case) and chest X ray (2 cases). The histological examination concluded to a bright cell carcinoma. In the former case, evolution was marked by spontaneous abortion at the fifth post-operative day, and the absence of metastasis after 16 months. In the second case, the evolution was worse because of metastasis occurrence four months after normal delivery. Pregnancy represents a privileged period for the discovery of renal tumor. Exams of tumor extension must consider pregnancy age. The treatment is similar to that of usual renal cancer and is associated to a simultaneous supervision of the pregnancy. PMID- 11859664 TI - [Pseudotumoral xanthogranulomatous pyelonephritis: diagnosis with percutaneous biopsy and success of conservative treatment]. AB - Focal xanthogranulomatous pyelonephritis is an unusual form of chronic renal infection that is difficult to diagnose prior to surgery. We report on a 19-year old woman who presented with a renal mass that mimicked malignancy. The diagnosis of focal xanthogranulomatous pyelonephritis was first suspected by radiological findings and further confirmed by histopathologic examination of percutaneous biopsy specimens of the lesion. Successful treatment of the patient was achieved with antibiotic therapy alone. Maximal efforts, including percutaneous renal biopsy, should be made to establish the diagnosis of focal xanthogranulomatous pyelonephritis before a therapeutic decision is reached. We recommend the use of antibiotics as a first-line treatment for patients with focal xanthogranulomatous pyelonephritis. PMID- 11859665 TI - [Arterio-ureteral fistula: diagnostic and therapeutic approach]. AB - The diagnosis of arterio-ureteric fistula must be considered in the case of sudden onset of abundant or intermittent haematuria occurring in a particular context (history of aorto-iliac vascular surgery, prolonged ureteric stenting, ilio-pelvic radiotherapy). Emergency treatment must control bleeding. Endovascular stenting may be a useful technique, followed by reconstructive surgery with vascular bypass graft and treatment of the ureteric lesion. PMID- 11859666 TI - [Klippel-Trenaunay syndrome and multiple vesical hemangiomas: treatment with Neodymium:YAG laser]. AB - The authors report the case of a patient presenting with bladder haemangiomas in the context of Klippel-Trenaunay syndrome treated by Neodymium:YAG laser. Klippel Trenaunay syndrome consists of a combination of hypertrophy of a limb, cutaneous angiomas and varicose veins. Bladder haemangioma is a benign congenital vascular tumour associated with Klippel-Trenaunay syndrome in 3 to 6% of cases, especially affecting children and young adults. Its most frequent clinical manifestation is haematuria. The diagnosis is based on endoscopy. Endoscopic treatment by Neodymium:YAG (Nd:YAG) laser photocoagulation appears to be a satisfactory treatment option. PMID- 11859667 TI - [Three cases of spontaneous intraperitoneal rupture of the bladder]. AB - Spontaneous rupture of the bladder is a rare disease which raises real diagnostic and therapeutic problems. Based on three new cases of spontaneous rupture of the bladder in the absence of any trauma and a review of the literature, the authors discuss the various diagnostic, prognostic and therapeutic aspects of this unusual lesion, secondary to multiple aetiologies, predominantly inflammatory causes. Idiopathic rupture of the bladder is particularly serious compared to other forms of traumatic or iatrogenic rupture. Late diagnosis caused by the absence of pathognomonic signs and the generally insidious clinical course at least partly contribute to this severity. Neoplastic forms have a very poor prognosis. PMID- 11859668 TI - [Intrauterine device migrating to the bladder]. AB - Transuterine migration of an intrauterine contraceptive device (IUCD) is a rare complication. The authors report a case of IUCD which migrated into the bladder and subsequently became calcified. This 34-year-old woman had been fitted with an IUCD for 4 years. She presented an unwanted pregnancy and signs of disabling cystitis at the end of pregnancy. The diagnosis of migration of the IUCD into the bladder with secondary calcification was suggested on the plain abdominal x-ray and on ultrasound and was confirmed by cystoscopy. Ballistic lithotripsy of the bladder stone with endoscopic extraction of the IUCD was then performed. PMID- 11859669 TI - [Wolfram syndrome. Report of 3 cases]. AB - Wolfram's syndrome is an autosomal recessive neurodegenerative disease. The major manifestations consist of diabetes mellitus and bilateral optic atrophy and are sufficient to establish the diagnosis. Urinary manifestations constitute an integral part of the syndrome, as they are present in up to 62% of cases and represent one of the major causes of morbidity. Three cases allow emphasize the difficulty of treatment, which is essentially symptomatic, and largely guided by the results of the various complementary investigations, especially urodynamic assessment. Although the causes of this syndrome are multifactorial and complex, it appears that the urinary lesions are at least temporarily improved by correction of excessive bladder pressure. PMID- 11859670 TI - [Brain metastasis of prostatic cancer: regression under hormonal treatment]. AB - Central nervous system metastases from prostate cancer are exceptional secondary sites, reported in less than 4% of postmortem cases. The authors describe an unusual case of cerebral metastasis from prostate cancer in a 44-year-old man that temporarily regressed during endocrine therapy. PMID- 11859671 TI - [Myxoid liposarcoma of the spermatic cord]. AB - A case of myxoid paratesticular liposarcoma in a 68-year old patient is reported. The evolution was normal during the follow-up period (22 months). Scrotal liposarcoma is a rare tumour: less than 100 cases were reported in literature. The diagnosis is difficult, clinical and radiological signs are the same as those of any intrascrotal tumour. Orchidectomy is the usual treatment, prognosis depends on the histological features, it is usually better than that of other paratesticular sarcomas. It did not need any additional treatment. PMID- 11859672 TI - [Transobturator urethral suspension: mini-invasive procedure in the treatment of stress urinary incontinence in women]. AB - Transobturator tape is an artificial tape designed for urethral suspension to treat female stress urinary incontinence. This tape has two original features: its non-woven polypropylene structure is coated with silicone on the urethral surface in order to limit retraction of polypropylene and to establish a barrier to extension of periurethral fibrosis. transmuscular insertion, through the obturator and puborectalis muscles, reproduces the natural suspension fascia of the urethra while preserving the retropubic space. A preliminary study (40 implantations) confirmed the feasibility of this operation, the low morbidity (one complication: sepsis) and the encouraging results between 3 and 12 months; in the treatment of isolated incontinence (16 patients), no postoperative dysuria has been observed; 15 patients are totally continent and 1 patient is improved; in the treatment of prolapse associated with frank or potential incontinence (24 patients), transient postoperative dysuria was observed in 4 cases, with no postoperative incontinence. PMID- 11859673 TI - [Deferred adjustment of the tension of tension-free vaginal tape (TVT) after surgical repair of stress urinary incontinence in women]. AB - INTRODUCTION: Urinary retention is a frequent complication of repair of female urinary incontinence treated by tension-free vaginal tape (TVT). The authors report a modification of the original technique designed to decrease the risk of postoperative dysuria and retention. MATERIAL AND METHODS: One hundred and twenty four patients with pure stress urinary incontinence were treated by TVT. In 74 patients (group 1), the TVT was placed according to the original Swedish technique. In 50 patients (group 2), no tension was applied to the suprapubic extremities of the TVT during the operation, but they were preserved and buried in an antiseptic dressing. On the day after the operation, traction was applied to the suprapubic TVT in the case of persistent incontinence. The immediate postoperative results and the results at 3 months, in terms of continence and urethral obstruction, were compared. RESULTS: Perfect continence was obtained in 84 +/- 6.5% and 94 +/- 4.2% of patients in groups 1 and 2, respectively (p = 0.08). Acute urinary retention (15% vs 2%, p < 0.03) and post-voiding residual urine > 50 cc (38% vs 10% on D1 and 14% vs 2% at 3 months) were more frequent in group 1. At 3 months, the reduction in maximum and mean flow rate was lower in group 2 (p < 0.03). In group 2, deferred traction of the TVT was necessary in 20% of cases, without causing any major infectious complications or pain. CONCLUSION: Deferred traction of TVT appears to decrease the rate of dysuria and urinary retention following repair of female urinary incontinence by tension-free vaginal tape (TVT). This technical modification does not affect the results on continence, which appears to be at least as satisfactory. This technical variant appears to be associated with a low morbidity. PMID- 11859674 TI - [Laparoscopic sacral colpopexy: an attractive approach for prolapse repair]. AB - OBJECTIVE: Evaluate laparoscopic sacral colpopexy. MATERIAL AND METHODS: 76 patients operated by laparoscopy between 1993 and 1999. The mean age of the patients was 59 years. 35 patients suffered from stress incontinence. A history of abdominal surgery was never a contraindication to laparoscopy. Hysterectomy was not systematically performed and the type of correction of urinary incontinence varied between suburethral sling, Burch procedure and TVT. The operating time ranged from 100 to 345 minutes with a mean of 199 minutes. Patients were discharged from hospital between the 2nd and the 10th postoperative day (mean: 4.9 days). Nine out of 10 patients operated (89.40%) were discharged before the 6th postoperative day. The results were satisfactory with correction of prolapse in 74 patients (96%). Only one intraoperative complication occurred in our series: a minimal injury of the colon complicated dissection of the posterior floor during resection of the pouch of Douglas. This wound was sutured during the same operating time. One conversion was required following sudden onset of oxygen desaturation during carbon monoxide insufflation. Laparoscopic prolapse surgery is used increasingly frequently and this study confirms that laparoscopic sacral colpopexy is effective, reliable and reproducible. PMID- 11859675 TI - [Koff's urethral mobilization: report of 26 hypospadias presenting a distal division of the corpus spongiosum]. AB - AIM: To evaluate the Koff urethral mobilization in 26 patients with distal division of the corpus spongiosum. MATERIAL AND METHOD: Twenty six hypospadias underwent Koff's urethral mobilization between January 1st 1999 and January 31st 2001. All cases were performed by the senior author. The mean age at surgery was 36.3 months (14 to 117 months). All cases had a distal division of the corpus spongiosum. RESULTS: The mean follow-up was 4.64 months (1-24 months): Four cases (15%) presented with a penile bleeding when the dressing was removed 4 days after the procedure. One of them went back to theatre to stop the bleeding. Five (19.2%) presented with a late meatal stenosis requiring a secondary meatotomy. One had a urethral fistula which disappeared spontaneously a few months later. All had a satisfactory cosmetic result. DISCUSSION AND CONCLUSION: These results are compared with other series published and it seems that a full penile urethral mobilization is a significant alternative procedure whose main advantage is to avoid the use of any non-urethral tissues to reconstruct the hypospadiac urethra. The selection of cases must be cautious to avoid secondary meatal stenosis which seems to be related to ischaemia of the distal hypoplastic urethra. PMID- 11859677 TI - [The York Manson approach in the treatment of prostato-rectal fistulae]. AB - The York Mason approach appears to be one of the most suitable techniques for the treatment of prostato-rectal fistulas, as it provides a maximum chance of success with no morbidity, as no cases of anal incontinence have been reported to date. The authors report a case of prostato-rectal fistula secondary to transurethral resection of the prostate for benign prostatic hyperplasia treated by the YORK MASON posterior trans-anosphincteric approach. PMID- 11859676 TI - [Tendinopathy associated with fluoroquinolones: individuals at risk, incriminated physiopathologic mechanisms, therapeutic management]. AB - The use of fluoroquinolones in urology has grown considerably over recent years. Unfortunately, although these molecules are not associated with severe life threatening complications, they have nevertheless been associated with tendon lesions responsible for functional disability. The frequency of these complications is probably underestimated. There is a variable lag-time (3 to 5 days) between introduction of the antibacterial and onset of pain. The symptom most frequently reported is pain over the tendon affected and the tendons most frequently affected are those submitted to high constraints. Bilateral lesions are present in 66% of cases. Although Pefloxacin is associated with the highest frequency of tendon complications (2.7% versus 0.2-0.3% for other fluoroquinolones), the duration of treatment appears to be important in every case, with a peak frequency after a fortnight of treatment. Although these complications were considered for a long time to be associated with patients presenting certain risk factors (age, steroid therapy, renal failure), they can also occur suddenly, in young adult sportsmen or non-sportsmen, with no known tendon disease. Several hypotheses have been proposed to explain the development of these cases of tendinopathy: immuno-allergic mechanisms, direct toxicity of the molecule on collagen fibres, cell-mediated oxidative aggression, or tendon necrosis due to vascular mechanisms. The outcome remains favourable in 75% of cases of tendinitis and in 49% of cases for tendon rupture. Contraindications must therefore be identified and the duration of treatment must be adapted, as the functional handicap can be long and particularly severe. PMID- 11859678 TI - [Balloon stent: simple and efficient drainage in hypospadias repair]. AB - The authors describe an original technique of hypospadias drainage with the combined advantages of balloon catheter and simple stent without the disadvantages of these modes of drainage. PMID- 11859679 TI - [The urologic work of Thomas Jonnesco (1860-1926)]. AB - Thomas Ionescu, eminent Rumanian surgeon and anatomist, was a great ambassador of french medicine. Politician and patriot, he was founder of modern rumanian experimental surgery and initiator of general rachi-anaesthesia. Among his innumerable scientific works we underline the urologic part of them and especially his original method of nephropexy. PMID- 11859680 TI - Male menopause may be real. PMID- 11859681 TI - Undertreating pain in women: a risky practice. PMID- 11859682 TI - Anticoagulant and thrombolytic therapies in women. PMID- 11859683 TI - Body image and eating behaviors in Orthodox and Secular Jewish women. AB - OBJECTIVE: To explore the impact of religion on the development of disturbances in body image and eating behaviors. PARTICIPANTS: 78 Orthodox Jewish women were compared with 48 secular Jewish women. DESIGN: Participants completed the Body Shape Questionnaire (BSQ), the Eating Disorder Examination-Questionnaire Version (EDE-Q), and the Figure Rating Scale (FRS). RESULTS: Despite a similar body mass index of 22.2 +/- 2.8 SDs, the secular women scored significantly higher on the BSQ (P = .005) and the EDE-Q (P = .004) than the Orthodox women. The secular women also had greater eating disorder symptomatology: more laxative use (P = .02) and a trend toward more vomiting (P = .06) and diuretic use (P = .06), although not more binge eating. They were twice as likely to have a fear of becoming fat (P = .05) and were four times as likely to be influenced by their shape and weight (P = .001). Also, despite increased media exposure, the secular group chose an ideal body size on the FRS similar to that of the Orthodox group, suggesting that their greater body dissatisfaction on the BSQ was related, instead, to greater cultural pressure for thinness (P = .007) and more shame about appearance (P = .04). CONCLUSION: Our findings show that membership in a strict, insulated religious group such as Orthodox Judaism may protect women, to some extent, from developing body dissatisfaction and eating pathology. PMID- 11859684 TI - Sex differences in drug metabolism: cytochrome P-450 and uridine diphosphate glucuronosyltransferase. AB - The two most common families of enzymes involved in drug metabolism are the cytochrome P-450 (CYP) and the uridine diphosphate glucuronosyltransferase (UGT) enzymes. The activity of these enzymes depends on genetic, physiologic, and environmental effects. Because the genes involved in the CYP and UGT proteins are not X-linked, the incidence of poor metabolizers would not be expected to be sex dependent. However, genetics also controls the amount (or activity) of the enzymes. Sex-dependent differences have been demonstrated for several CYP isoenzymes and for UGT. Ethnicity also appears to play a role in the activity of these enzymes. PMID- 11859685 TI - Gender differences in trauma and posttraumatic stress disorder. AB - BACKGROUND: Research on posttraumatic stress disorder (PTSD) has focused primarily on Vietnam War veterans. A handful of recent studies have been conducted on samples of the general population. OBJECTIVE: We sought to examine gender differences in exposure to traumatic events and in the emergence of PTSD following exposure in the general population. SUBJECTS: A representative sample of 2181 persons in the Detroit metropolitan area, ages 18-45 years. METHODS: Subjects were interviewed to assess history of traumatic events and PTSD using Diagnostic and Statistical Manual of Mental Disorders (Fourth Edition) criteria. The risk of PTSD was assessed in relation to a representative sample of traumas experienced in the population. RESULTS: Lifetime prevalence of traumatic events was slightly higher in men than in women. The risk for PTSD following traumatic experiences was twofold higher in women than in men. This gender difference was primarily due to women's greater risk of PTSD following events that involved assaultive violence. The probability of PTSD in women versus men exposed to assaultive violence was 36% versus 6%. Prior exposure to assaultive violence was associated with an increased risk of PTSD from a subsequent trauma; the gender difference in the vulnerability for PTSD is not explained by prior exposure. Duration of PTSD was longer in women than in men. CONCLUSION: The burden of PTSD in U.S. communities is greater in women than in men, chiefly due to the greater effect of assaultive violence on women. PMID- 11859686 TI - Gender differences in alcoholic cardiomyopathy. AB - The incidence, presentation, clinical features, and evolution of several cardiomyopathies have clear gender-related differences. In general, women show a different response to noxious cardiac agents than men, and they differ in myocardial adaptation to a variety of cardiac insults. Specifically in alcohol induced heart disease, women have shown different alcohol metabolism features and distinct pathophysiologic mechanisms leading to a higher sensitivity to alcohol induced heart damage. In preclinical alcohol-induced ventricular dysfunction, women were more sensitive to the toxic effects of ethanol than men. In overt alcoholic cardiomyopathy, women showed about the same prevalence of cardiomyopathy as men, despite having consumed far less ethanol. This supports a greater female propensity to alcohol-induced cardiac damage. PMID- 11859687 TI - Pregnant women may safely fly and exercise. PMID- 11859688 TI - The pharmaceutical company and the academic medical center: win-win or mutual exploitation? PMID- 11859689 TI - [Unexpected consequences of an expected adverse effect of cerivastatine]. PMID- 11859690 TI - [Differential diagnosis of systemic vasculitis: malignant rickettsiosis]. PMID- 11859691 TI - [Systemic lupus erythematosus in Martinique: an epidemiologic study]. AB - PURPOSE: To review epidemiological and clinical aspects of systemic lupus erythematosus (SLE) in Martinique, French West Indies. METHODS: Cases of SLE were identified by attending physicians. Patients who presented with at least four of the criteria defined by the American College of Rheumatology were included. Determination of incidence and prevalence included the new cases arising during the 1990-1999 period and 1999 population census results. Probability of survival was based on the use of the Kaplan-Meier estimator. RESULTS: Two hundred and eighty-six patients were studied, including 265 females (92.7%). The average annual incidence was 4.7 cases per 100,000 inhabitants (95% confidence interval [CI]: 2.5-6.9). The prevalence for 1999 was 64.2 cases per 100,000 inhabitants (CI: 56.2-72.2). The mean age at onset was 30 years. Eleven percent of all patients had at least one parent with SLE. Renal disease was present in 139 patients (48.6%), and neurological disorders were diagnosed in 70 patients (24.5%). Patients tested positive for the following antibodies: anti-Sm (37.1%), anti-RNP (58.7%), anti-SSA (47.2%). Mean survival time was: 96.4% (CI: 94.1-98.7) at 5 years, 91.8% (CI: 87.9-95.7) at 10 years. Survival was significantly reduced in patients with end-stage renal disease (n = 40, chi 2 = 6.96, P < 0.01). CONCLUSION: The high incidence of SLE in Martinique and the immunological characteristics of patients were found to be similar to those described in other populations of African descent. The frequency of renal disease and survival rates were identical to those reported in Caucasians. PMID- 11859692 TI - [Candida endocarditis: retrospective study in 12 patients]. AB - PURPOSE: Candida endocarditis are rare, with a poor prognosis. Actually, the principal problem concerns the growing incidence of nosocomial fungal infections. The objective of the present investigation is to assess a disease which risks becoming more pronounced in the future. METHODS: We have collected observations of Candida sp. endocarditis between 1985 to 1997 from three French university hospitals. RESULTS: Twelve of the observations fit the Duke criteria of acute endocarditis. Patients were eight men and four women, with a mean age of 46 years. An immunodepression was found in seven cases, and four patients were active drug addicts. Six had an underlying heart disease at risk to acute endocarditis. Candidemia risk factors were found in nine cases, with an average of 2.7 risk factors per patient. The fungal agents detected were Candida albicans (eight cases), C. tropicalis (one case), C. parapsilosis (two cases), and C. glabrata (one case). These vegetations were on aortic (seven cases), mitral (three cases), tricuspid valves (two cases) or in other areas (three cases), with multiple localizations (two cases). In three observations, vegetations were associated with myocardium abscesses. Eight patients had embolic complications, two had a cardiac insufficiency leading to death. The treatment was medical in all of the cases and combined with a surgical treatment in ten cases. The surgery was performed, on an average, 17 days after diagnosis, allowing seven surviving patients. Among them, five received a secondary prophylaxis and no recurrence was recorded. CONCLUSIONS: Prognosis remains severe because of the voluminous, friable and necrotic vegetations, which favor embolic migrations and which are not easily accessible to antifungals, which penetrate poorly into these vegetations. Therapy is based on a medical treatment combined with a valve replacement which needs to be done early on, and is followed by a relapse prevention which can occur several years after the initial episode. PMID- 11859693 TI - [Oropharyngeal colonization by Gram-negative bacteria in elderly hospitalized patients: incidence and risk factors]. AB - PURPOSE: In a prospective study, we determined the predisposing conditions and the course of aerobic Gram-negative bacilli oropharyngeal colonization during the hospital stay in patients older than 70 years. METHODS: We studied 116 patients admitted in our internal medicine unit. Medical characteristics were documented for each patient. Oropharyngeal swabs were collected on the first, third, and seventh days after admission, and then once a week until discharge. Positive culture was defined as the presence of at least one colony of Gram-negative bacilli. RESULTS: We collected 306 samples from the 116 patients. The mean hospital stay was 10.6 days. Fifty patients (43%) had at least one positive oropharyngeal culture. Thirty-three patients (30%) were colonized on admission, but this prevalence decreased during hospital stay: 20% on the third day and 14% on the seventh day. In an univariate analysis, prior administration of antibiotics (P < 0.01) and use of nonsteroid anti-inflammatory drugs (P < 0.05) were associated with colonization. CONCLUSION: Aerobic Gram-negative bacilli oropharyngeal colonization was transient during the hospital stay in the elderly. The use of nonsteroidal anti-inflammatory drugs and the previous treatment with antibiotics are possible risk factors for colonization. PMID- 11859694 TI - [Pulmonary hypertension associated with connective tissue diseases]. AB - PURPOSE: Pulmonary hypertension is a rare but well-known life-threatening complication of connective tissue diseases. The aim of this article is to analyse the available literature and to report the experience of a pulmonary vascular diseases centre about this complication. CURRENT KNOWLEDGE AND KEY POINTS: Scleroderma and its limited variant, the CREST syndrome (calcification, Raynaud phenomenon, esophageal dysmotility, sclerodactily, telangiectasia), is the most common connective tissue disease affected by pulmonary hypertension. Dyspnea is the main symptom and is frequently severe. Echocardiography is an excellent exam to detect pulmonary hypertension. However, right heart catheterization is necessary to confirm the diagnosis of pulmonary hypertension and to test vasoreactivity with a potent vasodilator such as nitric oxide. Pulmonary hypertension is less severe in patients with connective tissue diseases perhaps because of an earlier diagnosis. A significantly lower proportion of patients presents an acute vasodilator response, suggesting an early constitution of irreversible pulmonary vascular lesions. Continuous intravenous epoprostenol therapy seems to be less effective as compared with patients with primitive pulmonary hypertension and does not improve survival. So, we observed dramatic improvement in rare cases after immunosuppressive therapy. FUTURE PROSPECTS AND PROJECTS: New treatments with oral, subcutaneous or inhaled stable prostacyclin analogs or with an endothelin receptor antagonist are currently being evaluated. The role of immunosuppressive therapy has to be defined. PMID- 11859695 TI - [New Level III opioids of the World Health Organization]. AB - PURPOSE: The new opioids and the new galenic forms, now available in France, require an update in practitioners' knowledge. The purpose of the present study is to help those prescribing select the appropriate opioid and its galenic form for pain relief. CURRENT KNOWLEDGE AND KEY POINTS: Presentation of pharmacological properties of opioids (mechanisms, pharmacokinetics and pharmacovigilance). Presentation of indications, modes of prescription and use of main opioids for pain (especially cancer pain). Examples for calculating required drug dosage depending on the clinical situation and the route of administration. Symptomatic treatments of the main undesirable side effects of the opioids, and actions to be taken in the event of accidental overdose. FUTURE PROSPECTS AND PROJECTS: Oral morphine is the treatment first recommended for nociceptive pain insufficiently relieved by WHO level I and II analgesics. The new immediate release galenic forms allow morphine titration and the treatment of breakthrough pain. Transmucosal fentanyl, soon available in France, is recommended for breakthrough pain in patients already under opioid treatment: it gives more rapid relied starting after only 5 minutes and it only acts for a short time. Transdermal fentanyl is indicated for stable cancer pain. It is particularly suitable when oral and injectable morphine routes are not available, or for patients with severe constipation. Hydromorphone is the first opioid recommended in France for severe cancer pain when morphine resistance exists or uncontrolled side effects are present (opioid rotation). The new opioids and the new galenic forms widen the range of therapeutic possibilities. Their use is well codified for cancer pain and must still undergo clinical trials for chronic non-cancer pain. When correctly indicated, opioid selection provides a considerable advance in pain management. PMID- 11859696 TI - [Difficulties in diagnosis of human dirofilariasis outside known enzootic areas]. AB - INTRODUCTION: Human non-visceral dirofilariasis, a mosquito-borne disease of carnivores (dogs), is chiefly due to Dirofilaria repens in France and is well known in the Mediterranean basin. This zoonosis can be misdiagnosed in northern areas of France. We present and discuss two human cases diagnosed in Abbeville and Amiens (Picardy) by histological examination. EXEGESIS: The former case appeared as an axillary tumefaction occurring in a 41-year-old women 6 months after holidays in Montauban (southwestern France), the other as an intraorbital 'tumor' in a 53-year-old man who travelled for professional purposes in Central Europe and North America. Morphological, clinical, and epidemiological data of these human infections are discussed and the diagnostic features in tissue sections for species identification are reviewed. CONCLUSION: Increasing travel customs during the last decades favour the emergence of zoonotic parasites unusually in human hosts. Outside known enzootic areas, diagnosis is often delayed until pathological examination. In France, the incidence of human dirofilariasis has steadily increased and must be considered in the workup of cutaneous or intraorbital nodules. PMID- 11859697 TI - [Rheumatoid purpura in adults and parvovirus B19 infection: fortuitous association or parvovirus B19-induced vasculitis?]. AB - INTRODUCTION: Henoch-Schoenlein purpura has been reported to be associated with parvovirus B19 infection, particularly in children and rarely in adults. We report the case of a 42-year-old patient presenting with this association. EXEGESIS: A 42-year-old patient was admitted to our medical center because of lower limb purpura. Henoch-Schoenlein purpura diagnosis was confirmed on histological findings (kidney biopsy) and concomitantly parvovirus B19 infection was proved by serological test (IgM+). Association of Henoch-Schoenlein purpura and parvovirus B19 infection has already been described. However, none of the reported studies demonstrated clearly the link between these two diseases. With regard to this observation, we wonder about the systematic use of the parvovirus B19 serological test in patients presenting first Henoch-Schoenlein purpura. Indeed, parvovirus B19-induced vasculitis is habitually controlled with intravenous immunoglobulins. CONCLUSION: A prospective study should explore the link between Henoch-Schoenlein purpura and primary parvovirus B19 infection. Moreover, we should evaluate intravenous immunoglobulins' efficacy in Henoch Schoenlein purpura associated with active parvovirus B19 infection in order to improve the prognosis of this disease. PMID- 11859698 TI - [Febrile form of ankylosing spondylitis]. AB - INTRODUCTION: Ankylosing spondylitis is a chronic inflammatory disorder that is rarely associated with severe systemic manifestations. EXEGESIS: We report the case of a 29-year-old man presenting with an atypical ankylosing spondylitis. The patient presented with a high spiking fever, a marked weight loss, and an unusual increase of acute phase response markers. Such features are rarely associated with ankylosing spondylitis and the review of the few similar reported cases in the literature showed that this presentation is usually associated with a severe disease course, and an asymmetrical joint involvement that is unresponsive to nonsteroidal anti-inflammatory agents. CONCLUSION: Clinicians should be aware of this unusual presentation of ankylosing spondylitis, which should be included in the list of aetiologies of fever of unknown origin, to avoid useless diagnostic procedures. The severity of the disease course may lead to the consideration of a more aggressive treatment strategy. PMID- 11859699 TI - [Splenic rupture in infectious disease: splenectomy or conservative treatment? Report of three cases]. AB - INTRODUCTION: Spontaneous splenic ruptures are rare but life-threatening complications of infectious diseases. Splenectomy is still the treatment of choice but numerous recent reports have documented favorable outcome with conservative treatment. EXEGESIS: We report three cases of splenic rupture occurring respectively with infectious mononucleosis, P. vivax infection and dengue fever. Diagnosis, treatment and indications are reviewed, an approach to management is suggested. The study included three military men respectively aged 23, 24 and 35 years, admitted for acute abdominal pain in the left upper quadrant. The abdominal computed tomography confirmed partial rupture (splenic hematoma) in the first two cases, and an hemoperitoneum in the dengue case. Splenic ruptures can reveal or complicate an evolving infection. Rupture can happen spontaneously or as a result of trauma, which may be minor and unnoticed. The typical presentation is acute, but progressive forms are described. The diagnosis is made by ultrasound and CT scan. Splenectomy dogma tends to be supplanted by conservative treatment. Non-operative management can be successful if appropriate criteria and a long period of follow-up are applied in carefully selected cases. When an operative approach is selected, conservative surgical treatment is attractive. Splenectomy should be reserved for patients with uncontrollable rupture or with recurrent splenic bleeding. CONCLUSION: Spontaneous splenic rupture are uncommon in infectious diseases. A multidisciplinary management is necessary. A conservative treatment should be considered in selected, closely monitored patients. PMID- 11859700 TI - [Takayasu's disease in Morocco. Report of 47 cases]. AB - PURPOSE: Takayasu's disease is a chronic inflammatory arteritis involving large vessels in young women. We studied Moroccan patients to evaluate clinical, biological, radiological and evolution features of this disease in our country. METHODS: Forty-seven patients with Takayasu's arteritis were studied retrospectively between 1988 et 1999. RESULTS: In our series involvement of the aortic arch and its branches was more frequent than the abdominal aorta and its branches. Stenotic lesions of renal arteries were rare. Ultrasound was useful in the diagnosis and the monitoring of the disease. Treatment with glucocorticoids gave good results, with improvement in half of the patients and remission with stabilisation in 40% of cases. Tuberculosis occurred in 8.5% of patients. CONCLUSION: The use of ultrasound and computed tomography angiography is helpful for the diagnosis and monitoring of the disease progression. Glucocorticoids help to induce long remission in about 80% of treated patients. We don't find any relationship between Takayasu's arteritis and tuberculosis. PMID- 11859702 TI - [Suspicious adrenal glands]. PMID- 11859701 TI - [Efficacy of the sparfloxacin-ethambutol combination in a case of cerebral tuberculosis]. AB - INTRODUCTION: Treatment of cerebral tuberculomas demands the combination of rifampicin, isoniazid, pyrazinamide and ethambutol. The place of fluoroquinolones remains to be defined. EXEGESIS: A 36-year-old woman presented with a cerebral tuberculoma. Treatment with an antituberculous quadritherapy regimen was begun, but because of serious side effects, isoniazid and rifampicin weren't withdrawn during the fourth month. Sparfloxacin and ethambutol biotherapy was then started and continued for 12 months, which led to the disappearance of the cerebral lesion. CONCLUSION: Our observation proves that sparfloxacin is an alternative for cerebral tuberculoma treatment in case of intolerance or resistance to first line antituberculosis drugs. PMID- 11859703 TI - [Post-pregnancy osteoporosis. A case report]. PMID- 11859704 TI - [Mechanism of multidrug resistance caused by retinoic acid]. AB - OBJECTIVE: To investigate the regulatory mechanism of multidrug resistance(MDR) caused by all-trans retinoic acid (ATRA). METHODS: ATRA and IL-4 were used to treat human hepatoblastoma cell line (HepG2) cells. The proliferative activity, synthesis of alpha fetal protein (AFP), and cell cycle distribution of tumor cells were observed to evaluate the degree of cell differentiation. Flow cytometry and in situ hybridization were used to determine the expressing levels of p53, bcl-2, P-glycoprotein (P-gp) and c-jun and c-myc mRNA. MTT assay was used to evaluate the sensitivity of the tumor cells to chemotherapeutic agents. RESULTS: Both ATRA and IL-4 could induce the differentiation of HepG2 cells. ATRA treatment of the tumor cells led to drug resistance in chemotherapy (resistant factors: 1.6-3.1), and IL-4 increased the sensitivity of the tumor cells to antineoplasic drugs (reversal index: 4-17). The level of P-gp expression in ATRA treated cells was increased from 54.2% +/- 8.6% up to 98.5% +/- 1.4% (P < 0.01), but IL-4 markedly inhibited expression of P-gp down to 25.4% +/- 7.3% (P < 0.01). Both ATRA and IL-4 treatment could down-regulate c-jun and c-myc mRAN expressions. The level of p53 and bcl-2 expression could be up- or down-regulated by IL-4 treatment but they were unaffected by ATRA treatment. CONCLUSION: Degree of cell differentiation and level of c-jun and c-myc mRNA expression might not be related to change in drug sensitivity by inducing differentiation of HepG2 cells with ATRA and IL-4. Increased expression of P-gp caused by ATRA might be one of the factors up-regulating MDR. p53 (or bcl-2) might be involved in regulating the sensitivity of antineoplastic drugs by inducing differentiation. PMID- 11859705 TI - [Preparation and characteristics of human adherent natural killer cells induced by rhIL-15]. AB - OBJECTIVE: This work was to do preliminary study on the characteristics and preparation of rhIL-15 induced adherent human natural killer cells (A-NK). METHODS: Natural killer cells (NK cells) were first separated by centrifugation on Ficoll-Hypaque gradients, plastic adherence and nylon wool column adherence. Then, they were further purified by Percoll discontinuous density gradient centrifugation and T cell panning. Fluorescence-activated cell scan (FACScan) was applied to evaluate the natural killer cells and assess their degree of purification. Then, the purified NKs were incubated in the presence of rhIL-2 (6,000 U/ml) or rhIL-15 (6,000 U/ml) and changed into adherent NK cells. In the next step, the adherent kinetics, proliferation and cytotoxicity of A-NKs obtained by two different cytokines were analyzed by cell counting, MTT and reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: The adherent percentage, cytotoxity and proliferation of A-NKs generated in rhIL-15 culture were higher than those in rhIL-2. CONCLUSION: IL-15 is a better stimulator than IL-2 to induce adherent natural killer cells. PMID- 11859706 TI - [cDNA expression array in the differential expression profiles of p53 regulated genes in nasopharyngeal carcinoma and the human normal nasopharynx]. AB - OBJECTIVE: To compare the gene expression map of nasopharyngeal carcinoma (NPC) tissue with that of the control tissue by cDNA array and to discuss possible reasons of TP53 accumulation in NPC tissue. METHODS: After the hybridization of Atlas human cancer cDNA expression array 7742-1, analysis of Atlas arrays by means of AtlasImage 1.01a was carried out. Then, the results of array were verified by reverse transcription-polymerase chain reaction (RT-PCR). Gene expression alteration on the protein level was verified by immunohistochemistry. RESULTS: 134 of 588 tumor-related genes were upregulated and 88 downregulated. Thirteen of 32 p53-regulated genes showed differential expression with 11 upregulated and 2 downregulated. CONCLUSION: (1) p53 dysfunction exists in NPC tissues, (2) MDM2, p21 and Bax may be involved in the regulation of nasopharyngeal carcinoma cell growth. PMID- 11859707 TI - [Positive immunoglobulin A expression in human epithelial carcinoma cell lines]. AB - OBJECTIVE: In the present study, we try to further elucidate which class of immunoglobulin is expressed in epithelial carcinoma cell lines. METHODS: Three assays, immunohistochemistry, Western blot and enzyme-linked immunosorbent assay (ELISA) were used to detect the cellular protein extracts and the cultivated supernatant. RESULTS: Seven human epithelial carcinoma cell lines, MCF-7 (human breast carcinoma cell line), SW480 (human colon carcinoma cell line), MGC (human gastric carcinoma cell line), HeLa (human cervical carcinoma cell line), HNE2, CNE1-LMP1 and Tet-on-LMP1-HNE2 (three human nasopharyngeal carcinoma cells) expressed IgA protein in their cellular extracts and cultivated supernatants. CONCLUSION: Immunoglobulin is expressed in epithelium-derived carcinoma cell lines, which is in conflict with the traditional immunoglobulin theory. Our findings imply that the immunoglobulin expressed in epithelial carcinoma cells might play a role in the carcinogenesis of epithelium-derived carcinoma. Understanding the mechanism of immunoglobulin expression by cancer cells may possibly further facilitate the improvement techniques in cancer diagnosis, therapy or the mechanism of carcinogenesis. PMID- 11859708 TI - [Localization and analysis of 1A6 gene by dual-color fluorescence in situ hybridization on gastric carcinoma tissue and tumor cell lines]. AB - OBJECTIVE: To locate 1A6 gene on the chromosome and study its copy number by dual color fluorescence in situ hybridization (FISH) on three tumor cell lines and 22 gastric tumors patients. METHODS: The single-color FISH of small size biotin labeled cDNA cloned in plasmid was used to locate the 1A6 gene on one of the chromosomes in C group. Genomic DNA of 1A6 gene was screened by polymerase chain reaction (PCR) in PAC library according to the sequence. The florenscence of green and orange was incorporated into the target gene-1A6 cloned in PAC and chromosome-12-specific alpha-satellite repeat DNA by nick translation, followed by dual-color FISH. 1A6 gene was located in the metaphase chromosome of the normal peripheral lymphocytes. 1A6 gene and chromosome 12 copy numbers were analyzed on touch slide of gastric cancer tissue and cell lines. RESULTS: 1A6 gene was located in 12q23.2-23.3. The chromosome 12/1A6 signal ratio was 0.96 1.01 in breast, ovarian and gastric cancer cell lines. The ratio was 0.93-1.11 in gastric cancer tissue touch slides. The number of chromosome 12 is disomic (87.7%), triploid (7.4%) in BMI cell line; multisomic(100%), including pentasomic (67.6%) in SKOV3; the multisomic (83.1%), including trisomic (71%) in SGC823. There are highly disomic rate in 86. 4% (19/22) of patients. CONCLUSION: 1A6 gene is in 12q23.2-23.3. Neither 1A6 amplification nor deletion in gastric cancer tissue and three cell lines was found. Further study is needed for the understanding of chromosome 12 copy number variation in different cancer cell lines. PMID- 11859709 TI - [Gene marking of human colon cancer LoVo cells with enhanced green fluorescent protein]. AB - OBJECTIVE: The aim of this study is to obtain a human colon cancer cell line marked with enhanced green fluorescent protein (EGFP) gene. METHODS: The G1FP retroviral vector, carrying EGFP gene, was constructed. When combined with liposome-mediated transfection with cross infection, EGFP virus producer cells were generated. The human colon cancer LoVo cells were transduced with EGFP retrovirus. The integration and expression of EGFP gene were analyzed with polymerase chain reaction (PCR) and flow cytometry. RESULTS: The ecotropic EGFP virus was obtained by cocultivation of packaging cells for two weeks, while the amphotropic virus producer cells Am12/G1FP, expressing high-level EGFP, were generated through cross infection with a titer of 1.1 x 10(5) infectious particles/ml. The genetically modified LoVo/GFP cells gave off strikingly bright green fluorescence, in which the integration of EGFP provirus was confirmed by PCR analysis. CONCLUSION: The LoVo/GFP cells, which express high-level green fluorescence, may provide an useful tool for the study of micrometastasis of colon cancer in vivo. PMID- 11859710 TI - [In vivo and in vitro effects of selenium-enriched garlic on growth of human gastric carcinoma cells]. AB - OBJECTIVE: To compare the effects of selenium-enriched garlic (Se-garlic), garlic, Na2SeO3, and garlic + Na2SeO3 on growth of human gastric carcinoma cells. METHODS: In vivo and in vitro assays were carried out in the cultured human gastric carcinoma cell line MGC803 transplanted in the nude mice by cell count, flow cytometry, Western blot and neoplasm volume measurement. RESULTS: The inhibition effect of garlic was similar to that of Se-garlic, but Na2SeO3 was weaker than Se-garlic. Combination of garlic and Na2SeO3 was stronger than Se garlic. In flow cytometry assay, the proportion of G1 phase was increased after 24 hour treatment of Se-garlic, garlic and Na2SeO3 in the non-synchronized cells. However, the proportion of S phase was increased in the synchronized cells. The proportion of G2 + M phase was increased both in non-synchronized and synchronized cells on treatment with the combination of garlic and Na2SeO3. The amount of Cdk2-CyclinE complex and Cdk4-CyclinD1 complex was decreased in all treated synchronized cells in immunoprecipitation and immunoblot assays. The growth of MGC803 tumor in male Balb/c nude mice was inhibited by administration of 1.67% of Se-garlic (Se 2 micrograms/g) in diet with an inhibition rate of 29.92% in tumor weight. Treatment of either 0.83% of Se-garlic or 1.67% of garlic or 4.38 micrograms/g of Na2SeO3 (Se 2 micrograms/g) could not appreciably inhibit the growth of tumor. Wrapping of a number of monocytes around the tumor was induced in 62.50% of tumors in 0.83% of the Se-garlic group. CONCLUSION: In vitro, Se-garlic is able to inhibit the growth of MGC803 cell through action of garlic. In vivo, Segarlic is able to inhibit growth of MGC803 tumor in nude mice by being better than garlic and selenite. PMID- 11859711 TI - [Effect of transfected Cx43 gene on the gap junction intercellular communication and the human glioma cells proliferation]. AB - OBJECTIVE: To investigate the role of connexin 43 (Cx43) gene in the gap junction intercellular communication (GJIC) and the development of glioma so as to demonstrate the feasibility of using Cx43cDNA as the target of gene therapy for gliomas. METHODS: TJ905 human glioblastoma cell line without Cx43 gene expression was transfected with Cx43cDNA mediated by lipofectamine. Northern blot, in situ hybridization and immunocytochemistry were used for examination of Cx43mRNA and its protein examination; MTT assay and silver colloid stain were used to detect cell proliferation. TUNEL method was used to determine the cell apoptosis. Scrape loading dye transfer (SLDT) was used to monitor GJIC. RESULTS: The TJ905 transfectants resulted in dramatic upregulation of Cx43mRNA and its protein, with the GJIC of transfected glioma cells restored. Clones with high Cx43 expression also showed reduced proliferation in vitro. However, the cell apoptosis did not increase following the transfection with Cx43 gene. CONCLUSION: These results indicate that Cx gene and gap junction intercellular communication are of crucial importance in malignant glioma cell growth and development. Cx43 gene may be a target for gene therapy of gliomas. PMID- 11859712 TI - [Comparison of tumor suppressor gene deletion between squamous cell carcinoma and adenocarcinoma lung cancer in Chinese]. AB - OBJECTIVE: To prove the hypothesis that there may be different tumor suppressor genes (TSGs) involved in genesis of squamous cell carcinoma (SCC) and adenocarcinoma (ADC) in non-small cell lung cancer (NSCLC) in the Chinese. METHODS: Twenty-two microsatellite markers flanking to or within 13 identified or candidate TSGs were used to detect loss of heterozygosity (LOH) in these regions in the specimens of 28 SCC patients and 13 ADC patients. RESULTS: Our data demonstrated frequent allelic loss of FHIT, p53, IFNA, VHL and p16 both in SCC and ADC. Three genes, PRLTS, PTEN and p57, showed statistically significant differences in the LOH frequency between SCC and ADC. CONCLUSION: The comparison of LOH frequencies between SCC and ADC indicates possible correlation between specific tumor suppressor genes and each type of lung cancer. The results also suggest that more attention should be paid to the PRLTS gene, which may play an important role in the development of ADC. PMID- 11859713 TI - [Detection of malondialdehyde-DNA adduct level by 32P-postlabeling assay in normal human esophageal epithelium and esophageal squamous cell carcinoma]. AB - OBJECTIVE: To study whether the main malondialdehyde-DNA adduct (M1-dG) produced by lipid peroxidation is involved in the carcinogenesis of esophagus. METHODS: DNA samples were isolated from normal esophageal epithelium (n = 32) obtained by biopsy and esophageal squamous cell carcinoma specimens (n = 30) obtained by surgery. All tissue samples came from individuals living in Linxian, Henan, a high-risk area of esophageal cancer. Contents of M1-dG adducts were detected by 32P-postlabeling method. RESULTS: M1-dG adducts were detectable both in the normal and cancerous tissue samples. However, normal esophageal epithelial tissues exhibited significantly lower levels of M1-dG adducts (median 3.4, range 1.7/10(8)-55.4/10(8) nucleotides) than those found in esophageal cancer tissues (median 14.1, range 1.4/10(8)-59.0/10(8) nucleotides, P < 0.0001). The adduct levels were neither associated with gender, age, tobacco smoking status or genetic polymorphism in the CYP2E1, an enzyme participating in the oxidation of ethanol to form reactive free radicals. CONCLUSIONS: Our findings provide evidence that DNA damage, resulted from lipid peroxidation, can accumulate in the normal human esophageal tissue and reach relatively high level in cancer tissue which suggests that M1-dG adducts may be involved in the initiation and progression of cancer with its mutagenic and carcinogenic effects. PMID- 11859714 TI - [Glutathione-S-transferase M1 genotype in patients with hepatocellular carcinoma]. AB - OBJECTIVE: To study the glutathione-S-tranterase M1 (GSTM1) genotype in patients with hepatocellular carcinoma (HCC) with aflatoxin B1(AFB1) in a high risk region in Guangxi. METHODS: Specific GSTM1 primers and PCR technique were used for the detection of GSTM1 genotype using the peripheral leukocytes. A total of 379 samples was examined including 162 HCC patients whose lesions had been confirmed by pathology, 177 adults of local residents without cancer and 40 young men coming from other places with incidence of HCC. RESULTS: The frequency of GSTM1 null genotype in HCC patients was 102/162(63%), in local residents it was 92/177(52%) and in young men from outside it was 13/40(33%), which were significantly different. CONCLUSION: GSTM1 is one of the pivotal phase II detoxicated enzymes for AFB1. GSTM1 genetic deletion predisposes the individuals to HCC. However, the development of a HCC not only requires a genetic susceptibility, but also a AFB1 highly contaminated environment. The synergism of carcinogenic viruses such as HBV and HCV is also needed. These, when happen to be present together in an area in Guangxi, would lead to a high incidence of HCC. PMID- 11859716 TI - [Mutation BRCA1 gene in 186 breast cancer patients]. AB - OBJECTIVE: To analysis the mutation of BRCA1 gene in 186 breast cancer patients. METHODS: Polymerase chain reaction-single stranded conformational polymorphism (PCR-SSCP) and sequential were used in this analysis. RESULTS: 13/186 of patients showed mutation which comprised 7% of the total number of patients. In 8 patients, the mutation was observed in the intron splicing region and in 5 patients, the mutation was on the exon. CONCLUSION: Mutation analysis of BRCA1 gene is helpful in the determination of developmental potential, early diagnosis and gene therapy for breast cancer. PMID- 11859715 TI - [Colon cancer risk factors in Jiashan county, Zhejiang province, the highest incidence area in China]. AB - OBJECTIVE: To investigate the risk factors of colon cancer in the highest incidence area, Jiashan county, providing scientific basis for monitoring and prevention. METHODS: With a population-based case-control study, 109 colon cancer patients with 1:7 matched controls were interviewed with uniform questionnaires. Conditional logistic regression was used for univariate and multivariate analysis of colon cancer. RESULTS: Ten years before, high monthly pork consumption (OR = 1.608), high monthly animal oil consumption (OR = 1.391), psychological depression (OR = 9.887), negative emotion (OR = 1.723), lower digestive tract diseases (OR = 4.163), and history of colorectal cancer in the first degree relatives (OR = 3.421) were the main risk factors of colon cancer. At that time, high monthly vegetable consumption was a protective factor for colon cancer (OR = 0.422). CONCLUSION: Colon cancer in Jiashan county was related to dietary factors. The research results support the colon cancer etiological hypothesis of "fat-bile acid" and "deficiency of dietary fibres". Psychological depression, inheritance and lower digestive tract diseases were also correlated with colon cancer. PMID- 11859717 TI - [Arsenic trioxide in the treatment of advanced primary liver and gallbladder cancer]. AB - OBJECTIVE: To evaluate the effect and toxicity of arsenic trioxide (As2O3) in treating primary liver and gallbladder cancer. METHODS: Twenty-nine advanced primary liver cancer and 4 gallbladder cancer patients were treated with As2O3 injection only, 15 mg i.v. qd for 14-21 days and was repeated after 2 weeks. RESULTS: The overall response rate was 15.2%, 13.8% in primary liver cancer (PR 4, NC 21 and PD 4). It was 25.0% in gallbladder cancer (CR 1, NC 2, PD 1). The major side reactions were mild bone marrow suppression and hepatic functional damage. CONCLUSION: As2O3 injection is effective in treating primary liver and gallbladder cancer with mild side reactions. It is worth studying in the future. PMID- 11859718 TI - [Combined percutaneous ethanol injection through liver puncture and transcatheter hepatic arterial chemoembolization for hepatocellular carcinoma]. AB - OBJECTIVE: To study the therapeutic effects of transcatheter arterial chemoembolization (TACE) combined with beta-ultrasound guided percutaneous ethanol injection (PEI) at multiple points for hepatocellular carcinoma. METHODS: Eighty-seven patients with hepatocellular carcinoma were divided into two groups: Group A, 45 patients were treated with TACE only, group B, 42 patients were treated with TACE plus PEI. RESULTS: In group A patients, the 1-, 2-, and 3-year survival rates were 66.7%, 41.4%, and 21.4%, respectively. Only 26.1% of cancer specimens showed complete necrosis by pathologic examination in group A. In group B, the 1-, 2-, and 3-year survival rates were 97.1%, 85.71% and 65.71%, respectively, with complete necrosis in 81.8%. CONCLUSION: The therapeutic effects of transcatheter arterial chemoembolization combined with percutaneous multiple point ethanol injection through liver puncture is much better than that of TACE alone. PMID- 11859719 TI - [Implanted percutaneous continuous hepatic arterial infusion system for nonresectable liver metastasis]. AB - OBJECTIVE: To evaluate the efficacy of continuous hepatic arterial infusion of 5 fluorouracil (5-Fu)/calcium folinate (CF) through an implanted percutaneous arterial infusion system for nonresectable liver metastasis. METHODS: Sixty-eight patients with nonresectable liver metastasis received continuous hepatic arterial infusion of 5-Fu 500 mg/m2.d, CF 100 mg/m2.d and dexamethasone 10 mg/d1-5 via a drug delivery system (DDS) implanted percutaneously with the catheter introduced via the femoral artery or left subclavian artery for 5 days, once every month in combination with cisplatin and/or mitomycin-C, epirubicin and cytoxan as a multi drug protocol. RESULTS: The overall response rate (CR + PR) was 47.1% (32/68), with 2 patients giving CR and 30 PR. The response seemed to correlate with the intrahepatic tumor burden as 60.6% response rate was observed in intrahepatic tumor burden of less than 25% versus 34.3% response rate in those with more than 25% (P < 0.05). The median survival period was 15.2 months. The 1-, 2-year survival rates were 61.1% and 35.2% respectively in 54 patients who were followed. The therapeutic result was better in patients with liver metastasis originating from the gastrointestinal tract carcinoma than those from non gastrointestinal tract carcinoma (P < 0.05). In patients with liver metastasis originating from the gastrointestinal tract carcinoma, the median survival was 17.6 months, with 1-, 2-year survival rates of 68.4% and 39.5%. There was no severe toxic side effects or complications. The toxic reaction in liver, biliary and gastrointestinal tract was mild. CONCLUSION: Continuous hepatic arterial infusion of 5-Fu/CF and dexamethasone via a subcutaneous drug delivery system is able to prolong the survival and improve the quality of life in patients with nonresectable liver metastasis. PMID- 11859720 TI - [Relation between transcatheter arterial chemoembolization and time of death in patients with hepatic carcinoma]. AB - OBJECTIVE: To evaluate the relationship between transcatheter arterial chemoembolization (TACE) and time of death in hepatic carcinoma patients. METHODS: One hundred and ninety-five patients (male 188, female 7) with liver cancer underwent TACE with patients dying at different intervals after treatment. Dose of antitumor drugs, amount of iodized oil, liver function, size, recurrence of surgical resection, pattern of tumor, metastasis, presence of portal vein thrombosis, post TACE relapse, post TACE liver function and AFP, etc, altogether 15 variables were subjected to statistical analysis with the Cox's hazard proportional model. RESULTS: According to univariate and forward stepwise regression analysis, factors associated with significantly worse survival were diffused type, multi-nodular growth of tumor, and tumor embolus in the portal vein (P < 0.0001). Before death, most patients had symptoms of chest oppression, shortness of breath, ascites, abdominal distension, jaundice, hepatic failure and hematemesis. Causes of death were hepatic coma (27.2%, n = 53), hepatic failure (23.1%, n = 45), hemorrhage from the digestive tract (36.9%, n = 72), multiple organ failure (5.1%, n = 10) and others (7.2%, n = 14). CONCLUSION: Several factors of TACE may lead to failure in liver function and death. TACE may hasten death in patients with severe liver functional embarrassment. Before instituting TACE, careful weighing the pros and cons of the general condition and liver function is important for patients with portal vein embolism, multiple tumors or diffused lesions. Tolerance of liver to the drug, time, dose and method of treatment should be meticulously and carefully planned. Post TACE protection of liver is also very important. PMID- 11859721 TI - [Cox multivariate analysis of prognosis and proposal on a modified staging system of thymoma]. AB - OBJECTIVE: To investigate the clinicopathologic features, prognostic factors and propose a new modified staging criteria for thymoma. METHODS: The data of 159 patients operated for thymoma collected were retrospectively analyzed as to their prognostic factors and criteria of clinical staging through comparison of survival rates by the actuarial method, Log-rank and Cox multivariable model. RESULTS: Thorough resection (OR = 2.10), and extent of tumor invasion (OR = 1.73) were the most important prognostic factors. Tumor with peritumoral adhesion and absence of a complete capsule but without external invasion belonged to the criteria of stage II (II a). According to the prognosis and state of resection, lesions with obvious peritumoral invasion into the nearby organs but were thoroughly resected belonged to stage II (II b) and those incompletely resected belonged to III a. The unresectable lesions belonged to III b. CONCLUSION: According to the Cox prognostic multivariable model, the criteria of clinical staging of thymoma are defined as: Stage I-completely encapsulated non-invasive tumor without tight fibrous adhesions to the surroundings. Stage II a-only capsular invasion or capsule incomplete or with tight fibrous adhesions to the surroundings. II b-invasion into the surrounding fatty tissue, pleura, partial pericardium or lung, Stage III a-invasion into the great vessels, heart, trachea or hilus, which are resected with difficulty, or only palliatively. III b extensive invasion into the nearby organs, and resection is impossible. Stage IV a-pleural or pericardial dissemination, IV b-lymphatic or hematogenous metastasis. PMID- 11859722 TI - [Thymoma-report of 166 patients]. AB - OBJECTIVE: To discuss the characteristics of operation and the prognosis of 166 patients with thymoma. METHODS: 166 thymoma patients were treated from February 1985 to February 2000. By Masaoka staging system, there were 102 (61.4%) stage I, 28 (16.9%) stage II, 24 (14.5%) stage III, 12 (7.2%) stage IV a and 0 stage IV b lesions. The relation between stage and survival rate was analyzed. RESULTS: One (0.6%) patient died of the operation. 137 (82.5%) patients underwent radical operation. Thirty patients were lost to follow-up. With the life table method, the 10-year survival rate was 56.8%, with 79.8% for stage I, 51.6% for stage II, 33.5% for stage III and 0% for stage IV patients. CONCLUSION: Diagnosis of thymoma still depends on both clinical and pathological findings, which are correlated with stage. The principal treatment is to resect the tumor as completely as possible so as to relieve the symptoms and prolong the life of the patient. PMID- 11859723 TI - [Palliative irradiation combined with chemotherapy for lung cancer with brain metastasis]. AB - OBJECTIVE: To evaluate to effects and complications of palliative irradiation combined with chemotherapy for lung cancer with brain metastasis. METHODS: Eighty two non-small cell lung cancer with brain metastasis were treated, of whom 48 had extracranial metastasis simultaneously. All patients received chemotherapy (Teniposide and cis-platin) prior to and also after radiotherapy. The multiple intracranial metastasis patients received whole brain radiation therapy to a dose of 33-39 Gy/11-13 fx. The others who had single intracranial lesion received whole brain irradiation of 30 Gy/10 fx and a boost of 15 Gy/5 fx. RESULTS: The overall 1- and 2-year actuarial survival rates were 46.7% and 6.7%. For patients with brain metastasis and extracranial lesions, the 1- and 2-year survival rates were 32.4% and 4.4%. For patients with intracranial metastasis only, they were 61.2% and 8.8%, respectively (P = 0.003). The median survival of these two groups were 8 and 12 months. There was no significant correlation between the survivals rate and the histological type or the number of intracranial metastasis. No severe complications were observed during the treatment and follow-up. CONCLUSION: The regimen of combined chemotherapy with radiotherapy is effective for lung cancer with brain metastasis. The survival rate of patients with intracranial metastasis only is obviously higher than those with multiple extracranial lesions in addition to brain metastasis. PMID- 11859724 TI - [Internal strontium-89 radiotherapy for malignant bony metastasis]. AB - OBJECTIVE: This work was done to evaluate the indication, effectiveness, and side effects of internal radiotherapy with radioactive nuclide strontium-89 (89Sr) in patients with malignant metastasis in the bone. METHODS: Fifty-six patients with skeletal metastasis received this internal radiotherapy. The patients were observed and followed up with respect to pain control, lesion improvement and side effects. RESULTS: The overall effective rate of pain control was 76.8% with the effective rate of prostatic cancer and breast cancer higher than 80%. The lesions in 81.8% patients as assessed by SPECT imaging, were improved. The mild lowering of white cells, platelets and red cells was the main side effect. CONCLUSION: Internal radiotherapy with 89Sr is very useful for patients with malignant cancer metastasis in the bone. PMID- 11859725 TI - [Diagnosis and treatment of 31 patients with splenic tumors]. AB - OBJECTIVE: Splenic tumor, a rare disease, is scarcely mentioned in the literature. Here, we wish to present a clinical study on 31 such patients treated in the PLA General Hospital with emphasis on diagnosis and treatment. METHODS: Thirty-one splenic tumor patients were treated from January 1990 to December 2000. The clinical manifestations and image characteristics of splenic tumor, diagnosis and treatment were analysed. RESULTS: Male 19, female 12. Their ages ranged from 13 to 80 years with an average of 56. The symptoms and signs were non specific. All patients were first diagnosed by beta-ultrasonography to have single or multiple space-occupying lesions. Then, 19 patients received CT and 7 MRI scans. The diagnosis of seven patients suffering from splenic cysts were initially made definite by beta-ultrasonography among whom the lesions of 6 were confirmed by operation and pathology. The diagnosis of 9 patients with metastatic tumor was ascertained by combined imaging and clinical history. Finally, the lesions were confirmed by operation and pathology. Fifteen patients with primary splenic tumor were found to have primary space-occupying lesion by imaging: Except one patient who was diagnosed to have splenic lymphoma and not operated, all the other 14 patients underwent splenectomy with their lesions confirmed by pathology. Altogether, 25 of these 31 patients were treated by splenectomy with uneventful recovery without any operative mortality. Final pathologic examination disclosed that 20(65%) of 31 lesions were benign. CONCLUSION: Clinical history and imaging scans are important in the diagnosis of splenic tumors. The differentiation of benignity and malignancy is difficult before operation. Surgery is satisfactory in treating splenic tumors. PMID- 11859726 TI - [Single topotecan or in combination with other chemotherapeutic agents for 18 recurrent advanced ovarian cancer patients]. AB - OBJECTIVE: This study was designed to evaluate the efficacy and toxicity of topotecan hydrochloride, used singly or combined with other drugs in patients with recurrent advanced ovarian cancer. METHODS: Eighteen ovarian cancer patients who had received cytoreductive operation 1 to 3 times and 8 to 18 courses of chemotherapy but still failed in DDP-based(86.3%) and Taxol-based(32.7%) as well as combined DDP and Taxol-based(30.7%) chemotherapy, were divided into 3 groups: 1. Topotecan 1.2 mg/m2 q.d.i.v. for 5 consecutive days every 3 weeks for a total of 6 patients and 14 courses. 2. Topotecan 0.7 mg/m2 q.d.i.v. for 5 consecutive days and cis-platin 20 mg/m2 q.d.i.v. for 3 consecutive days over 4 weeks and 3. Topotecan 0.7 mg/m2 q.d.i.v. for 4 consecutive days and taxol 100 mg/m2 q.d.i.v. for one day as well as cis-platin 20 mg/m2 q.d.i.v. for 3 consecutive days totalling 12 patients and 27 courses. All together 18 courses, median 2.7 courses, were given. RESULTS: All 18 patients were evaluable. PR was achieved in 3 patients with a response rate of 16.7%. The main adverse effects were hematological toxicity, lasting for 3-27 days. Ten (55.6%) patients developed leucocytopenia Grade III-IV which occurred in 16(33.3%) courses. Six(33.3%) patients developed thrombocytopenia Grade III-IV which occurred in 8(16.7%) courses. Six(33.3%) patients developed RBC reduction Grade III which occurred in 8(16.7%) courses. Peripheral nerve inflammation was most serious in the non hematologic toxicity. CONCLUSION: Topotecan is effective in recurrent advanced ovarian cancer patients who have failed in DDP or/and taxol-based regimes. Topotecan combined with cisplatin and taxol would be effective even if single topotecan has failed. Careful evaluation before treatment is indicated. PMID- 11859727 TI - Cloning raises issues of feminism, role of science. PMID- 11859728 TI - Perspectives. Bush budget built on money-saving assumptions. PMID- 11859729 TI - The recruitment challenge facing the nursing profession. PMID- 11859730 TI - Unholy crisis. PMID- 11859731 TI - Portrait: Leon Kass. The president's philosopher. PMID- 11859732 TI - The munchies crackdown. The feds go after hemp in power bars and chips. PMID- 11859733 TI - [Different methods for assessing quality in health sector]. AB - AN OBLIGATION: The "management" of quality is an integral part of "medical" management today. The health centers and professionals have to prove their value and know-how through distinct signs of recognition. A METHOD FOR ASSESSING QUALITY: This paper defines the method of assessment of quality and specifies the typology of the persons, organisms, supervisors and clients of the French health system who have a personal or collective interest in judging the quality of health services. DIFFERENT POSSIBILITIES: Numerous existing methods for assessing quality are presented together with their use, implementation and the use made by the actors and consumers of the health system. Some of these assessment methods are formal, others informal, some are mandatory, others optional; some concern the general population, others the professionals. Although know-how is essential in health, making it known is strategic today. PMID- 11859735 TI - [Epidemiologic study of urinary calculi in Western France]. AB - OBJECTIVES: An epidemiologic study of urinary calculi (N = 1843) was conducted in Western France: distribution according to the main chemical compounds, age and sex. Comparison with the results of a study with national recruitment (N = 10,617) and a study with regional recruitment (N = 1774). METHOD: The study involved 1843 stones characterized beforehand by morphological analysis associated with infra-red spectrophotometry (FTIR). If analysis of the composition of the stones was carried out on the totality of calculi, studies related to age and sex included only 1583 cases. Comparison of percentages was made using chi 2 test. RESULTS: The composition in main compounds of calculi was comparable with the results of other studies; minor significant compounds presented great differences, raising the problem of interpretation of the infra red spectra of the latter. Hence, our work was directed towards the analysis of the major compounds and we showed, like most authors, that monohydrate calcium oxalate is predominant in male (46%) as well as in females (37%). Calculi average sex-ratio was 2.19 but dehydrated calcium oxalate sex-ratio was 4.42, suggesting that this compound is found mainly in men. Conversely, for the majority of phosphate stones, the sex-ratio was lower or equal to one, indicating that they predominate in women. Infectious calculi (particularly struvite calculi) appeared slightly more frequent in our population than in other studies, whereas the number of uric acid calculi was lower. This, however, remains to be confirmed. CONCLUSION: The population studied was not significantly different from the national population regarding lithiasis, except perhaps for uric acid and struvite calculi, despite specific regional differences in diet and the role of nutritional factors in lithogenesis. PMID- 11859734 TI - [Elevation of serum lactate dehydrogenase. Diagnostic, prognostic and evolutive values]. AB - OBJECTIVE: Determination of serum lactate dehydrogenase (LDH) levels is an usual practice. However, its place in the diagnosis process is not clear. We have collected serum LDH levels superior to 2-fold the normal rate and we tried to determine their diagnostics interest and, predictive and progressive values. METHODS: Retrospective study during 3 months in hospitalized adults. LDH levels were measured by spectrophotometry (Normal rate: 313-618 UI/L). RESULTS: 196 cases with LDH level elevations higher than 1236 UI/L were analyzed. The etiology of LDH level elevations were was benign in 60% of cases, malignant in 36% and, undetermined in 5%. There was no difference in between average values of LDH level average values of benign and malignant etiology (2708 vs 2842 UI/l). LDH rates and high LDH level elevations were not helpful for in the diagnosis process (a variety of 43 etiology was able to elevate increased LDH rates). In 45% cases, LDH level was 2 to 3-fold the normal rate; in 47.5% cases, 3 to 10-fold normal rate, and in 7.5% cases, superior to 10-fold normal rate. LDH elevations superior to 10-fold the normal rate were caused by benign etiology in 11 cases and malignant disease in 4 cases. A level superior to 10-fold the normal rate was not helpful in determining to determine the benign or malignant characteristics of the initial disease. However, LDH rate superior to 10 normal rate was a pejorative predictive criteria (hospitalization in intensive careunity in 73% of cases and mortality rate of 53%). During follow up of a neoplasia or malignant hemopathy follow up, several LDH measurements LDH level determinations were determined in for a small number of patients. LDH level normalizations is are attributable to efficientan effective treatment; LDH level elevations are associated with a therapeutic failure echappment. Evolution in LDH levels evolution was influenced by progression in neoplasia and malignant hemopathy evolution and also by various several treatments such as like blood transfusions, growth factors, radiotherapy and chemotherapy. CONCLUSION: LDH level elevation, however whatever its rate, don't seem to do not help in differentiating have interest to differentiate benign from malignant diseases. However, an elevation LDH elevation higher than 10-fold the normal rate is a pejorative predictive criteria, since because the mortality rate is superior toupper than 50%. During follow-up of in the neoplasia and malignant hemopathy follow up, so long as they are measured at distance from treatment, variations in LDH levels are a good marker of evolution, rate variations represent an evolutive marker conditionally the level determination would be realize remote several treatment. PMID- 11859736 TI - [The Munchausen syndrome: a diagnosis not to be forgotten in urology]. AB - INTRODUCTION: Among the manifestations of Munchausen's syndrome, "neurological" forms may exist. OBSERVATION: We present the case of a patient presenting with urinary retention following treatment for urinary incontinence. The patient had injected himself with infected urine collected from his catheter, in order to create septicemia. COMMENTS: Nephritic colitis, false gall stones, addition of fecal matter or of food and saliva in the urine or the bladder, neurogenic bladder and urinary infections have all been reported as possible manifestations of Munchausen's syndrome. Diagnosis of this syndrome is often delayed and laborious, after repeated hospitalisations, examinations and often aggressive treatment. Its therapeutic management is difficult. Patients, despite proof, deny their deceit and refuse psychiatric care. PMID- 11859737 TI - [Chemoprevention of colorectal cancer]. AB - A NEW CONCEPT: Chemoprevention of cancer consists in the administration of chemical agents to prevent or inhibit carcinogenesis. This strategy can be applied at any stage of carcinogenesis. ASSESSMENT: The development of such agents relies on classical bases: phases I, II and III. The approach consists in assessing the effect of the substance tested in patients with history of resected adenomas of the colon and at high risk of relapse and/or family risk of colon cancer. THE PRINCIPLE AGENTS UNDER ASSESSMENT: Are aspirin, type 2 cyclo oxygenase inhibitors, calcium, folic acid, certain vitamins, hormone replacement therapy for menopausal women and difluoromethylornithine (DFMO). PMID- 11859738 TI - [Endoscopic monitoring after polypectomy. What efficacy? What rhythm?]. AB - IN THE ABSENCE OF ENDOSCOPIC MONITORING: The risk of colorectal cancer is higher than in the general population in patients presenting an adenoma with a diameter of more than 1 cm, and/or comprising villous structures and/or severe dysplasia. The risk is not increased in the presence of one or two tubular adenomas measuring less than 1 cm and without severe dysplasia. It has been demonstrated that endoscopic monitoring decreases the incidence and mortality of colorectal cancers in these patients. MONITORING MODALITIES: A control colonoscopy, 3 years after polypectomy is sufficient in most cases. Colonoscopy is recommended within the 3 months following exeresis of a transformed sessile adenoma, in order to check that the polypectomy was complete. A control colonoscopy can be performed 1 year later in the rare cases presenting more than 3 adenomas, with one measuring more than 1 cm. When the control colonoscopy is normal, one can wait at least five years before conducting a further control. PMID- 11859739 TI - [The new chemotherapy of colorectal cancers]. AB - THE SITUATION: With around 35.000 new cases per year and a 40% mortality rate, colorectal cancer is a real problem of health care in France. Chemotherapy for metastatic disease has completely changed during the ten last years with the emergence of promising new drugs such as oxaliplatin and irinotecan. Oral 5-FU prodrugs have also shown an interesting efficacy in this setting. RESULTS: Showed increased response rate, progression free survival with both drugs, and overall survival, only with the irinotecan-based regimen. Moreover, after aggressive first-line chemotherapy, some patients can undergo surgical resection of the metastases, initially considered as unresectable. We still do not know if these aggressive first-line therapies should be proposed to all metastatic patients or only to a selected subgroup. The best strategies remain to be evaluated. IN THE SETTING OF ADJUVANT CHEMOTHERAPY: Of colorectal cancer, a 6-month 5-FU/folinic acid postoperative chemotherapy is actually the standard for the management of patients with positive lymph nodes (stage III). In stage II patients the benefit of such treatment remains controversial. The benefit of using oxaliplatin or irinotecan in association with 5-FU/folinic acid as adjuvant therapy is under investigation. CONCLUSION: In this 21st century, chemotherapy has a major place in the therapeutic management of colorectal cancer patients. PMID- 11859741 TI - Medicare carriers in S.C. restricting coverage of vitamin D. PMID- 11859740 TI - Medicare rate for Aranesp delayed. PMID- 11859742 TI - RPA/ASN position on the use of disease management in ESRD care. AB - The care of patients with chronic renal insufficiency and ESRD is at a major crossroad, and there is considerable uncertainty about the future. There are substantial risks for both the patients and the caregivers as well as opportunities to improve patient care and the milieu in which the care is provided. The treatment of ESRD and chronic renal insufficiency is moving into an era of collaborative care with RCMs, physician assistants, and advanced practice nurses predicated in large part on the anticipated shortage of nephrologists as the ESRD patient population continues to increase. The future of reimbursement is uncertain and payers may well have difficulty responding to new models of providing care. It is imperative that nephrologists accept the challenge and assume the leadership role as disease management and other collaborative methodologies of caring for our patients develop. PMID- 11859744 TI - Small improvements seen in patient, staff vaccinations. PMID- 11859743 TI - Guidelines on disposing of medical waste in the dialysis clinic. AB - The term "medical waste" varies from state to state as to its name, definition, and scope of coverage. In this article, we will focus on the process of how a dialysis clinic ensures proper classification, labeling, packaging, tracking, and disposal of medical waste. In addition, we will reference: OSHA regulations (29CFR1910), state specific regulations, DOT regulations (49CFR) and FDA regulations that impact the disposal of medical waste. PMID- 11859746 TI - NIDDK accepting requests for applications for a clinical trial in daily dialysis. PMID- 11859745 TI - The cost of home nocturnal hemodialysis in Ontario, Canada. PMID- 11859747 TI - Nutritional advantages of nightly home hemodialysis. PMID- 11859748 TI - The role of the renal social worker in the 21st century. PMID- 11859749 TI - Certification in advanced practice nephrology nursing: is it necessary or even desirable? PMID- 11859750 TI - Losing ground. A look at the recent decline in PD therapy. PMID- 11859752 TI - MedPAC to recommend a 2.4% composite rate increase. PMID- 11859751 TI - Technician training in Georgia: an educational approach. PMID- 11859753 TI - Surface plasmon resonance of calcium-binding proteins. PMID- 11859754 TI - Differential scanning calorimetry. PMID- 11859755 TI - Isothermal titration calorimetry. PMID- 11859756 TI - Multiangle laser light scattering and sedimentation equilibrium. PMID- 11859757 TI - Small-angle solution scattering reveals information on conformational dynamics in calcium-binding proteins and in their interactions with regulatory targets. PMID- 11859758 TI - Calcium binding to proteins studied via competition with chromophoric chelators. PMID- 11859759 TI - Investigation of calcium-binding proteins using electrospray ionization mass spectrometry. PMID- 11859760 TI - Synthetic calcium-binding peptides. PMID- 11859761 TI - Proteolytic fragments of calcium-binding proteins. PMID- 11859763 TI - Cadmium-113 and lead-207 NMR spectroscopic studies of calcium-binding proteins. PMID- 11859762 TI - Electron magnetic resonance studies of calcium-binding proteins. PMID- 11859764 TI - Calcium-43 NMR of calcium-binding proteins. PMID- 11859765 TI - Exploring familial relationships using multiple sequence alignment. PMID- 11859766 TI - Deconvolution of calcium-binding curves. Facts and fantasies. PMID- 11859767 TI - Structure determination by NMR. Isotope labeling. PMID- 11859768 TI - Protein structure calculation from NMR data. PMID- 11859769 TI - Shape and dynamics of a calcium-binding protein investigated by nitrogen-15 NMR relaxation. PMID- 11859770 TI - Quantitative analysis of Ca(2+)-binding by flow dialysis. PMID- 11859771 TI - The use of dipolar couplings for the structure refinement of a pair of calcium binding EGF domains. PMID- 11859772 TI - Vector geometry mapping. A method to characterize the conformation of helix-loop helix calcium-binding proteins. PMID- 11859773 TI - Use of calmodulin antagonists and S-100 protein interacting drugs for affinity chromatography. PMID- 11859775 TI - Gene expression in transfected cells. PMID- 11859774 TI - Enzymatic assays to compare calmodulin isoforms, mutants, and chimeras. PMID- 11859776 TI - Monitoring the intracellular free Ca(2+)-calmodulin concentration with genetically-encoded fluorescent indicator proteins. PMID- 11859777 TI - Studying the spatial distribution of Ca(2+)-binding proteins. How does it work for calmodulin? PMID- 11859778 TI - Absorption and circular dichroism spectroscopy. PMID- 11859779 TI - Fourier transform infrared spectroscopy of calcium-binding proteins. PMID- 11859780 TI - Steady-state fluorescence spectroscopy. PMID- 11859781 TI - Fluorescence methods for measuring calcium affinity and calcium exchange with proteins. PMID- 11859782 TI - How to keep your job in a tight market. PMID- 11859783 TI - Liability risks in clinical research. PMID- 11859784 TI - E&M: out with the new, in with the old. PMID- 11859785 TI - What's on the horizon for primary care? PMID- 11859786 TI - To keep your practice humming, get the right help. PMID- 11859788 TI - "I'm not a doctor. I'm an ophthalmologist". PMID- 11859787 TI - Doctors who go the extra mile. Fighting hunger and death half a world away. Interview by Robert Lowes. PMID- 11859789 TI - At Kaiser, a vision of the connected future. PMID- 11859790 TI - I'm a primary care physician--and proud of it! PMID- 11859791 TI - When your patient lies. PMID- 11859792 TI - [The influence of the vaccine prophylaxis and chemo-prophylaxis on the incidence of tuberculosis among children during the epidemic outbreak]. AB - The activities of a mass tuberculin diagnosis team improved with a phthisiopediatrician being included as its member. Team members' work quality standards have been developed: age-specific tuberculin sensitivity, the rates of infection with Mycobacteria tuberculosis in children aged 12-14 years were 29.3 +/- 2.9%; the annual risk of primary infection was 2.5 +/- 0.21%; the rate of hyperergy to tuberculin with qualitative drug prevention was 0.09%. This enhances the efficiency of BCG revaccination and drug treatment in Dispensary Registration Group 6 and stabilizes the incidence of tuberculosis in children with its rise in adults. PMID- 11859793 TI - [Microbiological blood studies in the diagnosis of tuberculosis in adolescents]. AB - Luminescence bacterioscopy of capillary blood has been found to be an additional rapid laboratory method that verifies a clinical diagnosis in cases of negative sputum bacterioscopic tests in 48.8% of the adolescent patients. The Boy luminescence microscopy shows that in adolescent tuberculosis patients, Mycobacteria are significantly more frequently present in the capillary blood (72.4%) than in the sputum (30.9%) and bronchial washes (34.8%). The detection of Mycobacteria in the capillary blood has been evidenced by cultural tests of some venous blood samples in the BACTEC 460 TB system. PMID- 11859794 TI - [Principles in the diagnosis of primary and occult manifestation of tuberculous infection in children]. AB - An algorithm of complex examination is proposed for early diagnosis of tuberculosis in children with different manifestations of tuberculous infection. The results of clinical and laboratory studies of 209 children by using the most informative values in its diagnosis may establish it in 81.8% of cases in the outpatient setting. While revealing high specific sensitization, some signs of functional disorders or 1-2 direct X-ray signs of an intrathoracic lymph nodal response were detected in 53.1% of the children with suspected tuberculosis, this condition may be interpreted as a borderline between Mycobacterium tuberculosis infection and disease. PMID- 11859795 TI - [Clinical and x-ray characteristics of respiratory diseases in children]. AB - Clinical and X-ray characteristics were studied in 102 children with first detected active respiratory tuberculosis. The patients with intrathoracic lymph nodal tuberculosis were 39.2%, those with a primary complex, tuberculosis pleurisy, disseminated tuberculosis, infiltrative tuberculosis, and focal tuberculosis, 33.3, 14.7, 4.9, 5.9, and 1.9%, respectively. Under the present conditions, respiratory tuberculosis is encountered more commonly (72.5%) in infants and pre-school children, which is characterized by its complicated course (30.4%), clinical symptomatology with signs of tuberculous intoxication (66.6%), hyperergic (29.4%) and pronounced (46.1%) tuberculin sensitivity. Surveys have revealed the disease in 58.8% of patients, by tuberculin diagnosis in 36.6 of them. PMID- 11859796 TI - [Specific features of tuberculosis in children having its different contacts with patients with active tuberculosis]. AB - The specific features of tuberculosis and its causes on different contacts were examined. Severe and complicated forms of tuberculosis were found to be diagnosed on all types of contact, which suggests that there is an epidemiological hazard of contact with not only bacteria-isolating person (47.1%), but also with a patient who has been found to have no Mycobacteria tuberculosis (15.6%), as well as with an unknown source of infection (37.3%). The causes of local tuberculosis irrespective of the type of contact were ascertained to be inadequate use of preventive measures for its detection in risk groups, untimely identification of infection sources, poor implementation of preventive measures in risk groups followed by a phthisiologist, disease-burdened premorbidity in children, as well as social troubles in their families. The paper shows it necessary to enhance preventive tuberculosis controlling measures in risk groups. PMID- 11859797 TI - [Comparative analysis of the results of tuberculosis questionnaire surveys among young persons]. AB - Sociosanitary monitoring evaluates the detected changes and predicts the health status of the population and its environment, establishes and eliminates its adverse human impact. Long-term sociological surveys for tuberculosis among the population of the Primorye Territory have revealed lower awareness in a young age group. Systemic antituberculosis propaganda, lectures with video films, discussions, questionnaire surveys in groups of young people will provide a more profound insight into the causes of tuberculosis, into the methods of its early detection and prevention. The results of questionnaire survey have been used in developing guidelines and differential programmes. PMID- 11859798 TI - [The role of specialized sanatoria in the prevention of tuberculosis in children]. AB - The paper presents the results of comparison of the rates of tuberculosis morbidity in children as a whole, in the foci of tuberculosis, in follow-up Group VI and the frequency of using sanatory care for preventive purposes in Russia in 1999. A role of available sanatoria in preventing tuberculosis is found to be in sufficient. By using the best areas as an example, the volume of required work with children at risk for tuberculosis and the rational use of sanatory care are substantiated. PMID- 11859799 TI - [Federal Law on "The prevention of the Spread of Tuberculosis in the Russian Federation]. PMID- 11859800 TI - [Time course of changes in tuberculin responses in children with monotonous tuberculin sensitivity on anti-tuberculous drugs]. AB - A group of 64 children with monotonous tuberculin sensitivity was analyzed. Intrathoracic lymph nodal tuberculosis (ITLNT) was revealed in 26 (41%) children. The remaining 38 (59% children with tuberculosis infection received a course of preventive treatment by taking into account the presence of risk factors. After tuberculosis treatment in children with ITLNT and tuberculous infection, papules due to Mantoux test significantly reduced in size. PMID- 11859801 TI - [Carbohydrate metabolism in long treated children and adolescents with respiratory tuberculosis]. AB - Carbohydrate metabolism was studied in 53 children and adolescents long treated for respiratory tuberculosis with preserving signs of its progression. The blood levels of cortisole, thyroxine, triiodothyronine were concurrently measured. Glucose tolerance impairments by the hypoglycemic type was found in 32.3 of the children and in 27.3% of the adolescents after long-term treatment for respiratory tuberculosis. Moreover, there was an increase (or an increasing trend) in the fasting plasma concentration of immunoreactive insulin. Latent carbohydrate metabolic disturbances are attended by high and low blood levels of cortisole and triiodothyronine, respectively. PMID- 11859802 TI - [Respiratory tuberculosis in old age children of the Republic of Sakha (Yakutia)]. AB - Analyzing epidemiological data and the authors' own findings has revealed the specific features of tuberculosis manifestations in old-age children. In this connection, the authors recommend that the scheme of tuberculosis controlling measures should be changed in children depending on their age. A differential tuberculin diagnosis scheme and a fluorographic examination of children above 13 years are proposed. PMID- 11859803 TI - [Post-tuberculosis changes in the lung and tuberculin sensitivity in risk group adults]. AB - There was a consecutive reduction in the incidence of residual changes in the lung and tuberculin hypersensitivity, which coincides with the periods of using different BCG vaccination methods in adults at risk for tuberculosis. In persons who were born in 1961-1962 or later when intracutaneous administration was used, these figures were 3.4 and 3.9 times lower than in those born before 1961-1962. The incidence of residual changes, tuberculin sensitivity in adult examinees were greater than the similar parameters in the general population. To detect and to follow up persons of these groups are pressing for stabilization of the epidemiology of tuberculosis. PMID- 11859804 TI - [Psychological peculiarities in adolescent patients with tuberculosis]. AB - The paper presents the results of psychological examination in 110 adolescent patients with respiratory tuberculosis and 89 healthy adolescents aged 13 to 17 years. Most adolescent patients were found to be highly anxious personalities, they were all diagnosed as having accentuation of character. The adolescent patients differed from healthy individuals by inadequate communicability, sensitive, liability to accumulation of negative emotions, by bad need for support and feelings to be taken hard, by marked internal strain and yearning for showing his/her individuality. This also had led to the lower behavioral range, made social adaptation difficult, promoted the susceptibility to stress exposure, and increased a risk for a disease. The psychological peculiarities found may be factors that predispose to tuberculosis. PMID- 11859805 TI - [Platelet function in patients with different stages of pulmonary tuberculosis]. PMID- 11859806 TI - [Informative value of polymerase chain reaction in the diagnosis of a tuberculous process]. AB - The informative value of the polymerase chain reaction (PCR) in the diagnosis of a tuberculous process in patients with pulmonary and extrapulmonary tuberculosis was studied. Sputum, urine, pleural and spinal fluids, and other specimens from 111 patients and 14 healthy individuals were tested. An analysis of sputum from 58 persons clinically diagnosed as having tuberculosis showed the following comparative sensitivity of the used methods: 82.8%, PCR; 65.5%, bacterioscopy; and 67.2%, inoculation. The detection rate of specific DNA from different samples was the following: 82.8% from the sputum, 66.6% from urine, and 54.5% from pleural fluid. The informative value of PCR in 14 patients with osteoarticular tuberculosis and in 39 with urinary tuberculosis was 42.8 and 40.6%, respectively. Thus, the application of PCR to a comprehensive examination of patients with tuberculosis of different sites is justifiable. PMID- 11859807 TI - [Dissolved ozone treatment-induced change in the resistance of multi-resistant mycobacterial strain to isoniazid and rifampicin]. AB - For 60 minutes, a mycobacterial (MBT) clinical strain resistant to streptomycin (S), rifampicin (R), isoniazid (I) was treated with dissolved ozone (PO3) at the concentration used for intravenous injection in the clinic. Then the strain was added to the Lowenstein-Jesen solid medium containing different concentrations of antituberculous agents. Following 3 weeks, drug sensitivity was determined by the number of grown colonies. Then MBT were retreated with PO3 in the same fashion, by repeating the cycle three times. At week 3, a growth of over 100 colonies was recorded in all control cultures. After each PO3 treatment of the strain, there was a reduction in its resistance to I and R. After triple treatment, MBT sensitivity to I completely recovered. In the R-containing media, there was also decrease in drug resistance, but the latter remained high (640 mu/ml). S resistance substantially lowered after the second PO3 treatment, but it restored after the third one. A mechanism responsible for lower MBT resistance to I and R under the action of "therapeutical" concentrations of PO3 is analyzed. The paper discusses whether MBT resistance can be changes at the phenotypic level rather that at the genetic one. PMID- 11859808 TI - [Polymerase chain reaction and ligase chain reaction in the diagnosis of gynecological tuberculosis]. AB - The polymerase chain reaction (PCR) and the ligase chain reaction (LCR) were evaluated and compared on 55 gynecological samples collected from women with active gynecological tuberculosis (Group 1), gynecological diseases other than tuberculosis (Group 2), and active tuberculosis elsewhere in the body without evidence for gynecological tuberculosis (Group 3). Acid fast staining and culture of Mycobacteria appeared to be ineffective in all specimens. The sensitivity of both amplification methods was 94.7% (one of the 19 patients with non tuberculosis). The LCx method showed a very high inhibition by specimens (64.1% of negative samples versus 16.7% for PCR and LCx could be useful tools in the diagnosis of gynecological tuberculosis. PMID- 11859809 TI - [A case of difficult differential diagnosis of disseminated pulmonary tuberculosis]. PMID- 11859810 TI - [Pulmonary actinomycosis]. PMID- 11859811 TI - [Chronic cough: pathogenesis and differential diagnosis]. PMID- 11859812 TI - [Accelerated culture diagnosis of tuberculosis by using automatic Bactec MGIT 960 and MB/BACT systems]. PMID- 11859813 TI - [Tuberculosis infection and incidence rates in children as an index of total tuberculosis epidemiological situation in Russia]. AB - The paper summarizes the results of the multidisciplinary epidemiological survey made in the past decade in the Russian Federation. Reasons for the higher incidence of tuberculosis among children and the influence of different factors on epidemiological indices were established. The paper clearly shows how the incidence and infection rates of tuberculosis in children are indicative of its total epidemiological situation in the general population in the country. Against this background, the pattern of new-onset tuberculosis mainly confirms the quality of work of general therapeutical and tuberculosis controlling services on active detection and prevention of the disease. PMID- 11859814 TI - [Tuberculosis in children: diagnosis of and methods for the evaluation of chemotherapy]. AB - The authors propose to determine altered forms of Mycobacteria, including sputum and blood granular forms, and the level of tuberculosis antibodies and serum Mycobacterium antigens by enzyme immunoassay as additional criteria for diagnosis of active primary tuberculosis in children. The diagnostic value of these indices is accounted for by the fact that they are found significantly more frequently and their level is significantly higher than the similar indices in healthy, tuberculosis-infected children and tuberculin-negative, prior BCG-vaccinated ones. By taking into account an decrease in the detection rate of sputum and blood granular mycobacteria and a change in the level of tuberculosis antibodies during chemotherapy, which are observed along with positive clinical and X-ray changes of a process, these tests should be recommended for evaluation of the efficiency of primary a bacillary tuberculosis in children. PMID- 11859815 TI - A grim outlook. Studies show shortage of healthcare workers grows. PMID- 11859816 TI - 'Inappropriate' gifts waning? Drug companies, and docs, too, rethink a practice. PMID- 11859817 TI - A new level of care. Providers that offer discounts make Blue Shield of California's A list. PMID- 11859818 TI - ViaHealth deal under fire. Court rules against system's plan for debt relief. PMID- 11859819 TI - Caring for a city of immigrants. PMID- 11859820 TI - Shades of AHERF. Debate since collapse of Enron revisits many issues raised in Pittsburgh healthcare system's high-profile bankruptcy. PMID- 11859821 TI - Capital spigot dripping faster. PMID- 11859822 TI - Too little or just right? Bush lays his healthcare budget on the table, but Democrats--and some Republicans--say $190 billion falls short. AB - In his State of the Union address last week, President Bush barely mentioned healthcare. But he tried to make up for it when he released his healthcare budget for 2003, which calls for $190 billion over 10 years to reform Medicare. The plan got a lukewarm reception from those in the industry, along with most Democrats and even some Republicans, who were hoping for a stronger stand. PMID- 11859823 TI - Murky water. IRS probes firm doing loan pools for hospital groups. PMID- 11859824 TI - [Declaration of the ethical principles for medical research in humans]. PMID- 11859825 TI - The slow, slow death of clinical engineering. PMID- 11859826 TI - How to create and maintain a Web site for your customers. Lessons learned from 3 experts in the field. PMID- 11859827 TI - How to stay motivated in your job. PMID- 11859828 TI - Endovascular management of a basilar artery false aneurysm secondary to endoscopic third ventriculostomy: case report. AB - OBJECTIVE AND IMPORTANCE: Third ventriculostomy for the management of noncommunicating hydrocephalus is a commonly performed procedure with a 5% complication rate. One of the known complications is basilar artery injury. CLINICAL PRESENTATION: We report a case of basilar artery injury, intraventricular hemorrhage, and false aneurysm formation in a 30-month-old boy after third ventricle floor fenestration. INTERVENTION: The false aneurysm was managed with endovascular trapping by use of Guglielmi detachable coils without morbidity. CONCLUSION: Endovascular therapy can be used successfully to manage vascular injury after third ventriculostomy. PMID- 11859829 TI - Anterior decompression of the atlantoaxial vertebral artery to treat bow hunter's stroke: technical case report. AB - OBJECTIVE AND IMPORTANCE: Bow hunter's stroke is a symptomatic vertebrobasilar insufficiency caused by stenosis or occlusion of the vertebral artery at the C1C2 level with head rotation. No case of anterior decompression of the vertebral artery for surgical treatment of bow hunter's stroke has been reported. CLINICAL PRESENTATION: A 47-year-old male patient presented with repeated episodes of unconsciousness caused by turning his head approximately 40 degrees to the right; he recovered consciousness within approximately 10 seconds after his head was returned to the neutral position. Angiography revealed an occluded right vertebral artery and temporary occlusion of the left vertebral artery, at the level of the C2 transverse foramen, when the patient's head was turned approximately 40 degrees to the right. INTERVENTION: Anterior decompression of the left vertebral artery at the transverse foramen of the axis was performed. Postoperative angiography demonstrated sufficient flow in the left vertebral artery even when the neck was rotated. CONCLUSION: The patient was discharged without neurological deficits. We demonstrate that simple surgical untethering of the vertebral artery at the transverse foramen of the axis is an effective method of treatment that avoids the limitation of head rotation. The advantage of this procedure is that it does not result in postoperative restriction of the patient's neck movements. The anterior approach, with decompression of the transverse foramen of the axis, in the present case provided adequate exposure of the vertebral artery and resulted in a satisfactory outcome. PMID- 11859830 TI - Anterior communicating artery aneurysms. PMID- 11859831 TI - Transcranial Doppler identification of changing autoregulatory thresholds after autoregulatory impairment. PMID- 11859832 TI - Image-guided transorbital roof craniotomy via a suprabrow approach: a surgical series of 72 patients. PMID- 11859833 TI - Image-guided transorbital roof craniotomy via a suprabrow approach: a surgical series of 72 patients. PMID- 11859834 TI - Intracranial metastatic parathyroid carcinoma: case report. PMID- 11859835 TI - Transcranial electrical motor evoked potential monitoring for brain tumor resection. PMID- 11859836 TI - Transluminal stent-assisted angioplasty of the intracranial vertebrobasilar system for medically refractory, posterior circulation ischemia: early results. PMID- 11859837 TI - High serum S100B levels for trauma patients without head injuries. PMID- 11859838 TI - High serum S100B levels for trauma patients without head injuries. PMID- 11859839 TI - High serum S100B levels for trauma patients without head injuries. PMID- 11859840 TI - Thrombocytopenia associated with clozapine and fluphenazine. PMID- 11859843 TI - [Sore tongue in a farmer]. PMID- 11859842 TI - [Problematic antihypertensive medication]. PMID- 11859841 TI - A tuned-trace theory of interval-timing dynamics. AB - Animals on interval schedules of reinforcement can rapidly adjust a temporal dependent variable, such as wait time, to changes in the prevailing interreinforcement interval. We describe data on the effects of impulse, step, sine-cyclic, and variable-interval schedules and show that they can be explained by a tuned-trace timing model with a one-back threshold-setting rule. The model can also explain steady-state timing properties such as proportional and Weber law timing and the effects of reinforcement magnitude. The model assumes that food reinforcers and other time markers have a decaying effect (trace) with properties that can be derived from the rate-sensitive property of habituation (the multiple-time-scale model). In timing experiments, response threshold is determined by the trace value at the time of the most recent reinforcement. The model provides a partial account for the learning of multiple intervals, but does not account for scalloping and other postpause features of responding on interval schedules and has some problems with square-wave schedules. PMID- 11859844 TI - Environmental radiological impact associated with non-uranium mining industries: a proposal for screening criteria. AB - Niobium, phosphate, coal and gold mining facilities have been selected as case studies with the aim of identifying possible sources of radiological impact during and after cessation of industrial operations. The results have shown that acid drainage, as well as chemical processing of mineral ores, constitute relevant impact indicators for present-day and future scenarios. The possible use of solid wastes abandoned at the end of the operations represents a long-term radiological concern. Therefore, it is necessary that Brazilian legislation for environmental licensing be revised taking into account the potential environmental and radiological impacts caused by these industries and establishing remedial procedures for waste storage areas in the existing sites. The indicators developed and procedures carried out can be used in screening for decisions on the adoption of regulatory requirements for practices at such types of installations. PMID- 11859845 TI - Molecular characterisation of a xyloglucan oligosaccharide-acting alpha-D xylosidase from nasturtium (Tropaeolum majus L.) cotyledons that resembles plant 'apoplastic' alpha-D-glucosidases. AB - We report the isolation, sequencing and analysis of the cDNA corresponding to an alpha-D-xylosidase involved in the mobilisation of xyloglucan from the cotyledons of germinated nasturtium (Tropaeolum majus L.) seeds. The translated open reading frame (2,808 bp including the stop codon), gave a polypeptide of 935 amino acids. It included the sequences of eleven peptides obtained by endo-proteinase digestion of the protein, and a putative hydrophobic signal sequence characteristic of a protein that is directed through the plasma membrane. The deduced molecular weight of the translated protein was appreciably higher than the molecular weight determined by sodium dodecyl sulphate-polyacrylamide gel electrophoresis, suggesting post-translational modification. The protein sequence showed high homology (76.0% identity over 896 amino acids) with a putative alpha xylosidase sequence from Arabidopsis thaliana and there was homology with several alpha-glucosidases, notably those associated with the plant cell apoplast. The enzyme is a member of Family 31 of the glycosyl hydrolases and it fits into Clade 1 of the phylogenic analysis of alpha-glucosidases. Although in vivo the nasturtium enzyme catalyses mobilisation of cell wall xyloglucan, the homology of its primary sequence with alpha-glucosidases prompted study of its action on a range of alpha-glucosides. It was active against several alpha-(1-->4)-and alpha (1-->6)-linked substrates, the former being hydrolysed faster. The functional and evolutionary relationships between this alpha-D-xylosidase and plant "apoplastic" alpha-D-glucosidases are discussed. PMID- 11859846 TI - The Antarctic psychrophile, Chlamydomonas subcaudata, is deficient in state I state II transitions. AB - State I-State II transitions were monitored in vivo and in vitro in the Antarctic, psychrophillic, green alga, Chlamydomonas subcaudata, as changes in the low-temperature (77 K) chlorophyll fluorescence emission maxima at 722 nm (F722) relative to 699 nm (F699). As expected, the control mesophillic species, Chlamydomonas reinhardtii, was able to modulate the light energy distribution between photosystem II and photosystem I in response to exposure to four different conditions: (i) dark/anaerobic conditions, (ii) a change in Mg2+ concentration, (iii) red light, and (iv) increased incubation temperature. This was correlated with the ability to phosphorylate both of its major light harvesting polypeptides. In contrast, exposure of C. subcaudata to the same four conditions induced minimum alterations in the 77 K fluorescence emission spectra, which was correlated with the ability to phosphorylate only one of its major light-harvesting polypeptides. Thus, C. subcaudata appears to be deficient in the ability to undergo a State I-State II transition. Functionally, this is associated with alterations in the apparent redox status of the intersystem electron transport chain and with higher rates of photosystem I cyclic electron transport in the psychrophile than in the mesophile, based on in vivo P700 measurements. Structurally, this deficiency is associated with reduced levels of Psa A/B relative to D1, the absence of specific photosystem I light-harvesting polypeptides [R.M. Morgan et al. (1998) Photosynth Res 56:303-314] and a cytochrome b6/f complex that exhibits a form of cytochrome f that is approximately 7 kDa smaller than that observed in C. reinhardtii. We conclude that the Antarctic psychrophile, C. subcaudata, is an example of a natural variant deficient in State I-State II transitions. PMID- 11859847 TI - Characterization of a mutant lacking carboxysomal carbonic anhydrase from the cyanobacterium Synechocystis PCC6803. AB - A fully-segregated mutant (ccaA::kanR) defective in the ccaA gene, encoding a carboxysome-associated beta-carbonic anhydrase (CA), was generated in the cyanobacterium Synechocystis sp. PCC6803 by insertional mutagenesis. Immunoblot analysis indicated that the CcaA polypeptide was absent from the carboxysome enriched fraction obtained from ccaA::kanR, but was present in wild-type (WT) cells. The carboxysome-enriched fraction isolated from WT cells catalyzed 18O exchange between 13C18O2 and H2O, indicative of CA activity, while ccaA::kanR carboxysomes did not. Transmission and immunogold electron microscopy revealed that carboxysomes of WT and ccaA::kanR were of similar size, shape and cellular distribution, and contained most of the cellular complement of ribulose-1,5 bisphosphate carboxylase/oxygenase (Rubisco). The ccaA::kanR cells were substantially smaller than WT and were unable to grow autotrophically at air levels of CO2. However, cell division occurred at near-WT rates when ccaA::kanR was supplied with 5% CO2 (v/v) in air. The apparent photosynthetic affinity of the mutant for inorganic carbon (Ci) was 500-fold lower than that of WT cells although intracellular Ci accumulation was comparable to WT measurements. Mass spectrometric analysis revealed that the CA-like activity associated with the active CO2 transport system was retained by ccaA::kanR cells and was inhibited by H2S, indicating that CO2 transport was distinct from the CcaA-mediated dehydration of intracellular HCO3-. The data suggest that the ccaA mutant was unable to efficiently utilize the internal Ci pool for carbon fixation and that the high-CO2-requiring phenotype of ccaA::kanR was due primarily to an inability to generate enough CO2 in the carboxysomes to sustain normal rates of photosynthesis. PMID- 11859848 TI - Endocrine aspects of healthy ageing in men. AB - Frailty is characterized by generalized weakness, impaired mobility and balance and poor endurance. Loss of muscle strength is an important factor in the process of frailty, and is the limiting factor for an individual's chances of living an independent life until death. In men, several hormonal systems show a decline in activity during ageing. Serum bioavailable testosterone and oestradiol, dehydroepiandrosterone and its sulfate, and growth hormone and insulin-like growth factor 1 concentrations all decrease during ageing in men. Physical changes during ageing have been considered physiologic, but there is evidence that some of these changes are related to this decline in hormonal activity. Studies on hormone administration in the elderly appear to be promising. However, until now, hormone replacement has not yet been proven to be beneficial and safe. PMID- 11859849 TI - Biochemical and sensory evaluation of wheat bran supplemented sorghum bread. AB - Addition of wheat bran to sorghum flour (Dabar cultivar) at two extraction rates 72% and 80% resulted in lowering reducing sugars. The percent decrease was 75.6% compared with the control at the end of fermentation period. There was a highly significant (p < or = 0.05) increase in crude fiber content as a result of addition of wheat bran. The increase was from 0.8 to 5.2 and from 0.5 to 5.3% for the 80% S/WB and 72% S/WB blends, respectively. Sorghum bread containing wheat bran was lower in reducing sugars and showed a significant increase (p < 0.05) in starch content. Sorghum bread containing wheat bran resulted in a lower in vitro protein and starch digestibilities. PMID- 11859851 TI - The factor VIII/von Willebrand factor ratio discriminates between reduced synthesis and increased clearance of von Willebrand factor. AB - It is often stated that a decrease in Von Willebrand factor (VWF), the carrier protein of factor VIII, results in a concordant change in factor VIII. Clinical data suggest that this is not always the case and we hypothesized that the ratio between factor VIII and VWF depends on the genetic defect that causes the VWF deficiency. We have analyzed the ratio between plasma factor VIII and VWF in several subtypes of Von Willebrand Disease and we show that the ratio is increased when VWF synthesis is reduced, but that the ratio remains one when VWF clearance is increased. These observations could be of clinical importance as an increased factor VIII/VWF ratio in combination with a borderline VWF level may indicate the presence of a true genetic defect, possibly a VWF null allele. PMID- 11859850 TI - Effects of second and third generation oral contraceptives and their respective progestagens on the coagulation system in the absence or presence of the factor V Leiden mutation. AB - Compared to second generation, the use of third generation oral contraceptives has been associated with an increased risk of venous thrombosis especially in women with the factor V Leiden mutation. To find an explanation for these risk differences we investigated the effects of desogestrel- and levonorgestrel containing oral contraceptives as well as their progestagens separately on the coagulation system in the absence or presence of the factor V Leiden mutation. In a single center, double blind trial, 51 women without and 35 women with the factor V Leiden mutation were randomized to either a second generation (30 microg ethinylestradiol/150 microg levonorgestrel) or a third generation (30 microg ethinylestradiol/150 microg desogestrel) oral contraceptive. After two cycles of use and a wash-out period of 2 menstrual cycles, the participants received the corresponding progestagen-only preparation containing 150 microg levonorgestrel or 150 microg desogestrel. In plasmas of the participating women fragment 1+2, factor V, VII, VIII, IX, X and XI were determined. Both combined oral contraceptives induced a decrease in factor V, whereas the levels of all other coagulant parameters increased. However, in women without the factor V Leiden mutation the effects of desogestrel-containing preparations were significantly different compared to levonorgestrel-containing oral contraceptives for factor V (-8.0; 95% CI -13.4 to -2.6), factor VII (26.8; 95% CI 15.5 to 38.0) and factor IX (-9.6; 95% CI -16.2 to -3.2). When these women used progestagen-only pills, a differential effect between desogestrel and levonorgestrel was only found for factor IX (-6.5; 95% CI -11.4 to -1.5). In carriers of the factor V Leiden mutation desogestrel-containing oral contraceptives induced more pronounced changes in factor V (-14.2; 95% CI -22.4 to -6.0) and factor VII (36.1; 95% CI 19.7 to 52.6) compared to levonorgestrel-containing oral contraceptives. Comparing desogestrel- and levonorgestrel-only, only for factor V a differential effect was found in these women (-9.5; 95% CI -18.3 to -0.6). It appears that desogestrel-containing oral contraceptives have a more pronounced effect on the coagulation system than levonorgestrel-containing oral contraceptives which may be explained by a less effective compensation of the thrombotic effect of ethinylestradiol by desogestrel. PMID- 11859852 TI - Field study of lyophilized calibrant plasmas for fresh plasma INR determination. AB - An alternative approach to INR estimation is for laboratories to calibrate their own local system using calibrant plasmas supplied by manufacturers or reference laboratories. The purpose of the present study was to investigate the within laboratory variability of a calibrant plasma procedure using various sets of lyophilized plasmas. INR had been assigned to 13 calibrant plasmas in a previous multi-center study. Each of 10 other ("field") laboratories measured PTs in the 13 calibrant plasmas and in 15 local fresh coumarin plasmas, using three different thromboplastin reagents. Each fresh coumarin PT was converted to INR using a calibration procedure with a set of 4 calibrants (1 normal + 3 abnormals). The abnormals of each set were either coumarin or artificial and were used with different assigned INR. When the INR had been assigned with a thromboplastin brand identical to the thromboplastin in the field laboratory, the procedure was named "reagent-specific" calibration. Otherwise the procedure was named "dissimilar" calibration. Using "reagent-specific" calibration procedures, relatively homogeneous INR were obtained for the fresh coumarin plasmas, whatever type of calibrant was used. In contrast, discrepant INR were obtained when "dissimilar" cross-species calibration procedures were used. The study was limited to 9 laboratories using the same type of coagulometer and one using a different type. PMID- 11859853 TI - Improved fibrinolysis after one year of treatment with enalapril in men and women with uncomplicated myocardial infarction. AB - AIMS: To study long-term effects of enalapril on mass concentrations of tissue plasminogen activator (tPA), plasminogen activator inhibitor (PAI-1), tPA/PAI-1 complex and von Willebrand factor (vWF) in both genders with uncomplicated myocardial infarction. METHODS AND RESULTS: More than three months after an uncomplicated myocardial infarction 82 survivors (46 males, 36 females) were randomised to enalapril/placebo. PAI-1, tPA, tPA/PAI-1-complex and vWF were measured after two weeks, six and 12 months following randomisation. PAI-1 decreased significantly in both genders in the enalapril-treated group after two weeks, with a maximum decrease at six months (mean reduction: 31% equal to 9.8 microg x L(-1), CI: 5.2 to 14.5 microg x L(-1), p = 0.0001) and remained significantly lower at 12 months. Mass concentration of tPA decreased significantly (mean reduction: 1.81 microg x L(-1), CI: 0.903 to 2.708 microg x L(-1), p < 0.001) after two weeks treatment in both genders but returned to baseline values at 12 months. The tPA/PAI-1-complex decreased and was significantly lower (mean reduction 0.96 microg x L(-1), CI: 0.36 to 1.56 microg x L(-1), p = 0.003) in the enalapril group after two weeks and six months (p = 0.037). No decrease of vWF was seen in the enalapril group. CONCLUSIONS: Enalapril treatment up to one year depressed mass concentrations of PAI-1 and transiently tPA and tPA/PAI-1 complex indicating an improvement of the fibrinolytic balance in both genders with uncomplicated myocardial infarction. PMID- 11859854 TI - The impact of the Kozak sequence polymorphism of the glycoprotein Ib alpha gene on the risk and extent of coronary heart disease. PMID- 11859855 TI - Sympathetic modulation of nitrergic neurogenic vasodilation in cerebral arteries. AB - The presence of close apposition between the adrenergic and the non-adrenergic or nitrergic nerve terminals in large cerebral arteries in several species is well documented. The axo-axonal distance between these different types of nerve terminals is substantially closer than the synaptic distance between the adventitial nerve terminals and the outermost layer of smooth muscle in the media. This feature suggests that a functional axo-axonal interaction between nerve terminals is more likely to occur than that between the nerve and muscle. Thus, transmitters released from one nerve terminal may modulate release of transmitters from the neighboring nerve terminals, resulting in a neurogenic response. We have reported that nicotine-induced nitric oxide (NO)-mediated neurogenic vasodilation is dependent on intact sympathetic innervation in porcine and cat cerebral arteries. Evidence also has been presented to indicate that nicotine acts on alpha7-nicotinic receptors located on sympathetic nerve terminals, resulting in release of norepinephrine which then diffuses to act on beta2-adrenoceptos located on the neighboring nitrergic nerve terminals to release NO and therefore vasodilation. The predominant facilitatory effect of beta2-adrenoceptors in releasing NO is compromised by presynaptic alpha2 adrenoceptors located on the same nerves. Activation of cerebral sympathetic nerves may cause NO-mediated dilation in large cerebral arteries at the base of the brain. PMID- 11859856 TI - Effect of protease-activated receptor-2 deficiency on allergic dermatitis in the mouse ear. AB - To investigate the involvement of protease-activated receptor-2 (PAR-2) in allergic dermatitis, we generated PAR-2-deficient (PAR-2(-/-)) mice. Ear thickness, contact hypersensitivity (CH) induced by topical application of picryl chloride (PC) or oxazolone (Ox) after sensitization, and vascular permeability after ear passive cutaneous anaphylaxis (PCA) were compared between wild-type (WT) and PAR-2(-/-) mice. Ear thickness was almost the same in untreated WT and PAR-2(-/-) mice. Topical application of PC or Ox thickened the ears at 6, 24 and 48 h after challenge with a peak at 24 h in WT mice. In PAR-2(-/-) mice, the ear swelling induced by both PC and Ox was suppressed at every time point, and significant inhibition was found at 24 h in PC-induced CH and at 24 and 48 h in Ox-induced CH. Histopathological observation of the ears at 24 h after challenge revealed that PC- or Ox-induced ear edema and infiltration of inflammatory cells in WT mice were greatly attenuated in PAR-2(-/-) mice. The vascular permeability in the ears after PCA was not different between WT and PAR-2(-/-) mice. These results strongly suggest that PAR-2 plays a crucial role in type IV allergic dermatitis but not in type I allergic dermatitis. PMID- 11859857 TI - Highly active palladium/activated carbon catalysts for Heck reactions: correlation of activity, catalyst properties, and Pd leaching. AB - A variety of palladium on activated carbon catalysts differing in Pd dispersion, Pd distribution, Pd oxidation state, and water content were tested in Heck reactions of aryl bromides with olefins. The optimization of the catalyst (structure-activity relationship) and reaction conditions (temperature, solvent, base, and Pd loading) allowed Pd/C catalysts with very high activity for Heck reactions of unactivated bromobenzene (turnover number (TON) approximately 18000, turnover frequency (TOF) up to 9000, Pd concentrations down to 0.005 mol %) to be developed. High Pd dispersion, low degree of reduction, sufficient content of water, and uniform Pd impregnation are criteria for the most active system. The catalysts combine high activity and selectivity under ambient conditions (air and moisture), easy separation (filtration), and quantitative recovery of palladium. Determination of Pd in solution after and during the reaction, and catalyst characterization before and after the reaction (transmission electron microscopy (TEM), X-ray diffraction (XRD)), indicate dissolution/reprecipitation of palladium during the reaction. The Pd concentration in solution is highest at the beginning of the reaction and is a minimum (< 1 ppm) at the end of the reaction. Palladium leaching correlates significantly with the reaction parameters. PMID- 11859858 TI - Optimising stereoselectivity in intramolecular Diels-Alder reactions of pentadienyl acrylates: synthetic and computational investigations into the "steric directing group" approach. AB - Experimental conditions for the intramolecular cycloaddition of four related pentadienyl acrylates 3, 4, 5 and 6 are reported. In contrast with several previous reports, pentadienyl acrylates do undergo synthetically useful intramolecular Diels-Alder reactions: 3, 4, 5 and 6 cyclise at reasonable rates at temperatures of 132-180 degrees C at atmospheric pressure in moderate to good yields. The stereochemical outcome of each of these reactions was accurately measured and the results are in good agreement with transition structure populations predicted using B3LYP/6-31+G(d) theory. The parent system 3 cyclises with moderate endo selectivity; the presence of either a C5-methyl substituent or a C3-bromine atom results in a slight shift towards the trans-fused exo stereoisomer but--overall--a less selective reaction. The presence of both C3-Br and C5-CH3 substituents results in a marked improvement in stereoselectivity with the exo,lk-product predominating. Interpretation of B3LYP/6-31+G(d) transition structures allows insights into the improvement in stereoselectivity obtained by incorporating a removable "steric directing group" into a 5-methyl-1,3,8 nonatriene precursor. PMID- 11859859 TI - Complementary and alternative medicine: how safe is it? PMID- 11859860 TI - Cure of testicular germ cell cancer: an index of access to healthcare. PMID- 11859861 TI - Puzzling epidemiological situations in Africa. PMID- 11859863 TI - UCLA PET in Alzheimer's diagnosis gets nation's attention. PMID- 11859862 TI - Nuclear medicine, molecular imaging, and molecular medicine. PMID- 11859864 TI - Children in accident and emergency: seen but not heard? AB - This study examined the ideas and beliefs about the care of children in Accident and Emergency (A & E) among a group of experienced nurses. The study was set in a single A & E department, during a time when a new paediatric area was being developed. Semi-structured interviews were used to generate qualitative data. Seven main themes emerged from the data, namely: priority of children in A & E; the role of the paediatric trained nurse in A & E; fear of dealing with children; staffing issues; a separate area for children; isolation; and motivation of staff. Further discussion of these issues, in the context of existing literature, confirmed the inherent difficulties in dealing with children in a predominantly adult environment. PMID- 11859865 TI - Convulsive status epilepticus in children with intractable epilepsy is frequently focal in origin. AB - BACKGROUND: Convulsive status epilepticus (CSE) is a common neurological emergency. Our objectives were to study children with recurrent nonfebrile CSE to assess the evidence for focal origin. METHODS: Series of 18 children with recurrent CSE and intractable epilepsy were identified by chart review. Clinical, radiological, and EEG data were reviewed. Focal structural abnormalities were identified on MRI and CT images by one neuroradiologist who was unaware of the clinical details. RESULTS: The patient's ages ranged between 6-22 years (mean 15.3, SD 4), and 67% were males. Most children (89%) had a severe cognitive and / or behavioural disorder. Most patients (89%) had multiple seizure types and 95% of these were partial seizures. Twelve (67%) children had at least one episode of CSE with focal features identified clinically. Focal brain abnormalities were detected on 18% and 55% of CT and MRI films respectively. Overall, 53% had a focal abnormality on structural neuroimaging. Interictal EEG revealed focal or multifocal abnormalities on at least one occasion in 94% and 22% of patients respectively. Overall, 17 patients had focal features on at least one EEG. Thirteen ictal EEGs were recorded on 11 (61%) patients. Ten (91%) of these recordings revealed a focal onset. CONCLUSIONS: Many handicapped children with recurrent CSE have focal clinical, radiological, or electrographic features. This supports a focal origin for CSE in most children with intractable epilepsy. PMID- 11859867 TI - In-vitro study of bacterial adherence to different types of intraocular lenses. AB - The aim of this study was to determine the adherence of Staphylococcus epidermidis to intraocular lenses made of five different biomaterials: polymethylmethacrylate (PMMA), heparinized PMMA, silicone, hydrophilic acrylic, and hydrogel. The extent of bacterial binding was measured by counting. The results were compared using a one-factor variance analysis. Adherence was weakest on hydrogel and strongest on the silicone polymer. Bacterial adherence to the implant surface must therefore depend on the hydrophobicity or hydrophilicity of the biomaterial. PMID- 11859866 TI - Structure-activity relationships (SAR) of contraceptive progestogens studied with four different methods using calculated physicochemical parameters. AB - Structure-activity relationships (SAR) of the contraceptive progestogens for (I) oral contraceptive activity (OCA), (II) androgenic effect, and (III) binding affinity for sex hormone binding globulin (SHBG) were studied using four different methods: principal component analysis (PCA), hierarchical cluster analysis (HCA), neural networks (NN), and electronic indices method (EIM) employing descriptors calculated by the semi-empirical Austin Model I (AM1) method. An additional set of molecules was used to check the reliability of the results obtained for OCA by PCA. Using PCA, three different sets of descriptors were found to correlate with the three different biological activities, I-III, indicating that the interaction between the receptor and the progestogen must depend on the type of biological activity. The descriptors selected by PCA were also employed for SAR analysis of the contraceptive progestogens using two other methods, HCA and NN. Both HCA and NN correctly classified high activity molecules as different from low activity ones. Thus, those descriptors selected by PCA work well in the other two methods of classification. Using the sign of p, a difference of electron densities of selected molecular orbitals in a specified region in a molecule, it was possible to discriminate high activity molecules from low activity molecules in the three different types of activities studied, I III, with one exception. PMID- 11859868 TI - Articular manifestations after the administration of intravesical BCG. PMID- 11859869 TI - Phenylketonuria and glycogen storage disease type III in sibs of one family. AB - Hyperphenylalaninemia result from a block in the conversion of phenylalanine into tyrosine due to a defect in either the enzyme phenylalanine hydroxylase (98% of subjects) or in the metabolism of the cofactor tetrahydrobiopterin. Phenylalanine hydroxylase deficiency is the most common form of inherited hyperphenylalaninemia disorders, with a prevalence between 1/4,000-1/40,000. Glycogen storage disease (GSD) type III is caused by debranching enzyme deficiency of glycogen degradation. The clinical features vary in relation to the localization of the enzyme defect. Two clinical entities exist: a combined hepatic myogenic form (GSD IIIa) and a purely hepatic form (GSD IIIb). The inheritance is autosomal recessive. We describe a Turkish family in which two girls were found to have phenylketonuria, while in two other sisters glycogen storage disease type III was diagnosed. The parents of these children are cousins and they have had 12 children. PMID- 11859870 TI - Loss of BRCA1 expression in sporadic male breast carcinoma. AB - BRCA1 protein is normally expressed in the nuclei of breast epithelial cells and functions as a negative regulator of the cell cycle. Mutations of BRCA1 gene have been associated with familial breast carcinomas. Although somatic mutations have not been demonstrated in sporadic breast carcinomas, loss of nuclear expression of BRCA1 protein has been correlated with high-grade, infiltrating breast carcinomas in females. We have investigated the expression of BRCA1 protein in a series of 26 infiltrating carcinomas of male breast and correlated the results with a variety of clinicopathological parameters. Immunohistochemical analysis demonstrated decreased expression of BRCA1 in all carcinomas compared to normal breast epithelium. The median survival was 8 years in patients with minimal loss of BRCA1 expression while it was 1.5 years in patients with marked loss of this protein. Marked loss of BRCA1 protein was also associated with increased cell proliferation. These results suggest that BRCA1 plays a similar role in both male and female breast carcinoma and Joss of this protein is associated with poor prognosis. PMID- 11859871 TI - Involvement of breast epithelial-stromal interactions in the regulation of protein tyrosine phosphatase-gamma (PTPgamma) mRNA expression by estrogenically active agents. AB - BACKGROUND: Protein tyrosine phosphatase gamma (PTPgamma) has been implicated as a tumor suppressor gene in kidney and lung cancers. Our previous results indicate that estradiol-17beta (E2)-induced suppression of PTPgamma may play a role in mammary tumorigenesis. Zeranol (Z), a nonsteroidal growth promoter with estrogenic activity that is used by the US meat industry, induces estrogenic responses in primary cultured breast cells and breast cancer cell lines. METHODS: PTPgamma mRNA expression in human breast tissues and cells isolated from surgical specimens of mammoplasty and breast cancer patients were detected and quantified by RT-PCR. Immunohistochemical staining was used to localize PTPgamma in human breast tissues. Breast epithelial and stromal cells were isolated and co-cultured to determine the involvement of cell-cell interaction in the regulation of PTPgamma mRNA expression by E2 and Z. RESULTS: PTPgamma mRNA expression was lower in cancerous than in normal breast tissues. Both E2 and Z suppressed PTPgamma mRNA levels in cultured normal breast tissues by approximately 80%, but had a lesser effect in cultured epithelial cells isolated from normal breast tissues. In the co-culture system, both E2 and Z suppressed PTPgamma mRNA to a greater degree in epithelial cells than in stromal cells. In whole breast tissues, PTPgamma was immunolocalized to the epithelium. Treatment with E2 or Z diminished PTPgamma staining indicating reductions in PTPgamma at the protein level. CONCLUSIONS: The results indicate that both E2 and Z regulate PTPgamma expression in human breast and that epithelial-stromal cells interaction is important in the regulation of PTPgamma expression by estrogenically active agents. PMID- 11859872 TI - Melatonin inhibits estrogen receptor transactivation and cAMP levels in breast cancer cells. AB - We have previously demonstrated that the pineal hormone, melatonin, can inhibit the growth of estrogen receptor-alpha (ERalpha)-positive breast cancer cells and suppress ERalpha gene transcription. To investigate the relationship between the estrogen response pathway and melatonin's growth inhibition, ERalpha-positive MCF 7 human breast cancer cells were transiently transfected with an estrogen response element (ERE) luciferase reporter construct and then treated with melatonin (10(-9)-10(-6) M) for 30 min followed by 10(-9) M 17-beta-estradiol (E2) or treated with each compound alone. Melatonin pre-treatment significantly reduced E2-induced ERalpha transactivation and ERalpha-ERE binding activity. We also conducted experiments to determine if melatonin modulates cAMP levels in MCF 7 cells. Melatonin inhibited the forskolin-induced and E2-induced elevation of cAMP levels by 57 and 45%, respectively. These data indicate that melatonin can act as a biological modifier to affect ERalpha transcriptional activity by regulating signal transduction pathways which impinge on the ERalpha and by altering E2-mediated ERalpha transactivation and ERalpha DNA binding activity. PMID- 11859873 TI - Dominant negative interference of transcription factor AP-2 causes inhibition of ErbB-3 expression and suppresses malignant cell growth. AB - ErbB-3 (HER3) is a member of the epidermal growth factor receptor family. Increasing evidence suggests that elevated expression of ErbB-3 is important for malignancy. In this study, we found that elevated levels of ErbB-3 expression did not occur in the absence of AP-2gamma in a panel of human mammary epithelial and fibroblasts cell lines. In contrast, there was no association between the expression of AP-2alpha or AP-2beta and the level of ErbB-3, or between AP-2alpha and AP-2gamma double positivity and ErbB-3 expression. In co-transfection experiments, exogenous expression of AP-2gamma robustly activated ErbB-3 promoter activity. Moreover, expression of a dominant negative AP-2 protein, AP-2delta (deleted residues 31-117), not only repressed the ErbB-3 promoter activity but also suppressed endogenous ErbB-3 transcription in the ErbB-3 overexpressing cell line MRC-5VA. Overexpression of AP-2A resulted in a decreased proliferation rate and inhibitin of colony formation. Taken together, these data strongly support a role for the AP-2 gene family, in particular, AP-2gamma, in the control of ErbB-3 expression. Interference with the function of transcription factor AP-2 might provide a potential strategy for modulation of the malignant phenotype. PMID- 11859874 TI - Organochlorines, p53 mutations in relation to breast cancer risk and survival. A Danish cohort-nested case-controls study. AB - Epidemiological studies integrating genetic susceptibility with biological measurements of organochlorine exposure may provide new clues regarding these substances influence on breast cancer etiology. Initial attempts pursuing this avenue has dealt with polymorphisms in the carcinoge metabolizing enzymes cytochrome P450 (CYP1A1). This study examined if mutations in the tumor suppressor gene p53 affected organochlorine exposure related breast cancer risk and survival. The material consisted of 162 breast cancer cases and 316 matched controls, who had participated, in the Copenhagen City Heart Study (CCHS) between 1976 and 1978. Cases diagnosed between study initiation and 1993 were identified by linkage to the Danish Cancer Registry. The case group served as a cohort in the survival analyses. Information on known and suspected breast cancer risk factors was obtained from CCHS, and the Danish Breast Cancer Cooperative Group provided information on tumor characteristics. Lipid adjusted serum concentrations of selected organochlorines were compared between cases and controls while stratifing by p53 mutation status. A non-significant increased risk of breast cancer was observed in the highest exposure level of dieldrin and polychlorinated biphenyls among women who developed a tumor with mutant p53 (odds ratio (OR) = 3.53, 95% confidence interval (CI) = 0.79-15.79 and OR = 3.00, 95% CI = 0.66-13.62). There was no clear difference in overall survival between breast cancer cases with 'wild-type' and mutant p53, although a significant dose response relationship appeared for dieldrin exposure in tumors with 'wild-type' p53. These preliminary results suggest that p53 mutations may have a modifying effect on at least the breast cancer risk associated with exposures to organochlorines. PMID- 11859875 TI - Prognostic value of epidermal growth factor receptor (EGFR) and its relationship to the estrogen receptor status in 1029 patients with breast cancer. AB - An epidermal growth factor receptor (EGFR) has been reported to be associated with a poor clinical outcome in breast cancer, while its prognostic value remains controversial. Immunohistochemical staining for EGFR was performed on frozen sections of primary breast cancer from 1029 patients with a mean follow-up duration of 46 months. EGFR was positive in 277 (26.9%) of 1029 cases which inversely correlated with the estrogen receptor (ER) status. A univariated analysis indicated that EGFR had a significant prognostic value in both the disease free survival (DFS) and the overall survival (OS), while the same effect was also found in node negative as well as node positive breast cancer. A multivariate analysis indicated that EGFR was an independently significant prognostic factor for DFS (p = 0.0174) and OS (p = 0.0105) in all patients, but that EGFR demonstrated a prognostic significance only for DFS (p = 0.0241) in node negative and only for OS (p = 0.0333) in node positive breast cancer. When all patients were stratified for EGFR and ER, a multivariate analysis indicated that the combination of EGFR(+)/ER(-) was an independently significant factor for both DFS and OS in node negative as well as node positive breast cancer. In conclusion, the prognostic value of EGFR was demonstrated by a multivariate analysis in a large series of breast cancer patients, but the value of EGFR was somewhat insufficient to achieve statistical significance for both DFS and OS in the subgroups divided by nodal status. On the other hand, the prognostic value of combination of EGFR and ER was sufficient to achieve statistical significance based on a multivariate analysis for both DFS and OS in the subgroups of node negative as well as node positive breast cancer patients. PMID- 11859876 TI - Overexpression of MDM2 oncoprotein correlates with possession of estrogen receptor alpha and lack of MDM2 mRNA splice variants in human breast cancer. AB - To evaluate the significance of murine double minute 2 (MDM2) oncoprotein in human breast cancer as a nuclear-cytoplasmic shuttling protein, an estrogen receptor (ER) alpha regulator, and a prognostic marker and to study how MDM2 is overexpressed, we investigated its status in tissue samples and examined the correlation between overexpression and MDM2 gene abnormalities, status, and clinicopathological parameters. We detected MDM2 oncoprotein in both nucleus and cytoplasm by frozen-section immunohistochemistry. There was a significant correlation between MDM2 overexpression and low-grade nuclear atypia, absence of lymph node involvement, and increased levels of ER alpha protein. Our molecular assays found no point mutations in Ser17, but there was a correlation between MDM2 overexpression and the lack of splice variant mRNAs. These results suggest that the distribution of MDM2 reflects its nuclear-cytoplasmic shuttling ability; that interaction between p53 and MDM2 for tumor progression is not enhanced by point mutations at codon 17; and that the expression of MDM2 splice variants is a reason for the lack of its overexpression. MDM2 overexpression correlates with favorable prognostic parameters. A decreased level of MDM2 will lead to a deviation from the ER alpha signaling pathway. PMID- 11859877 TI - Preservation of histopathologic assessment by use of a special technique for tissue inactivation by radiofrequency. AB - Thermoinactivation is under investigation as a new method of cancer treatment. Prior histopathologic verification has been indispensable. Our experiments show that thermoinactivation is in principle possible without destroying histopathologic morphology, if the temperature stays within certain limits. It could thus become applicable already before diagnostic biopsy, which might be used to avoid any potential spread during biopsy and before definitive surgery. PMID- 11859878 TI - Inhibition of myc-dependent breast tumor formation in transgenic mice. AB - One of the most promising approaches for cancer gene therapy is the use of the so called suicide genes, which encode prodrug-activating enzymes and render transduced cells more sensitive to prodrugs. The enzyme nitroreductase (NTR) converts prodrug CB1954 into a cytotoxic DNA interstrand cross-linking agent. We have established transgenic mice in which the pro-oncogene c-myc and NTR were fused to the internal ribosome entry site and coexpressed in luminal cells of the mammary gland under the control of mouse whey acidic protein (WAP) promoter to evaluate NTR mediated ablation of mammary tumors. More than 78% of transgenic females developed in situ or infiltrating carcinomas after three to four pregnancies. By contrast, if the transgenic female mice were given the prodrug CB1954 during their third lactation, the incidence of tumors decreased to less than 40% (P < 0.05). The total number of carcinomas was even more striking with 117 carcinomas identified in 14 non-ablated transgenics compared with only five in 15 treated animals (p < 0.05, student t test). C-myc induced pleomorphic nuclei and mitotic figures were seen as a field change in over 70% of the untreated transgenics compared to 20% in the treated group. Our results suggest that the enzyme pro-drug system NTR-CB1954 efficiently inhibit myc-dependent tumor formation and malignant progression in the mammary gland. PMID- 11859879 TI - Tuberculosis and treatment with infliximab. PMID- 11859880 TI - Tuberculosis and treatment with infliximab. PMID- 11859881 TI - Tuberculosis and treatment with infliximab. PMID- 11859882 TI - Tuberculosis and treatment with infliximab. PMID- 11859883 TI - Research in developing countries. PMID- 11859884 TI - Dreams, brick, and mortar: John Sharpe Chambers and the origins of the University of Kentucky Medical Center. PMID- 11859885 TI - On cloning: advocating history of biology in the public interest. AB - Cloning - the process of creating a cell, tissue line or even a complete organism from a single cell - or the strands that led to the cloning of a mammal, Dolly, are not new. Yet the media coverage of Dolly's inception raised a range of reactions from fear or moral repulsion, to cautious optimism. The implications for controlling human reproduction were clearly in the forefront, though many issues about animals emerged as well. On topics of public interest such as cloning, historians of biology have the opportunity to make a unique contribution. Such debates are often aired as if they have no precedents, either in biology or in the ethical, moral, and social concerns arising in the public arena. The technology leading to Dolly draws on strands of research going back to the 1890s, and the cycle of the public response has been repeated often in the past century. What can we learn from examining these events historically, and how can we - or should we even try - to inform public opinion? I think we should try and will outline briefly some of the ways that can work. PMID- 11859886 TI - The philosophical origins of Mitchell's chemiosmotic concepts: the personal factor in scientific theory formulation. AB - Mitchell's formulation of the chemiosmotic theory of oxidative phosphorylation in 1961 lacked any experimental support for its three central postulates. The path by which Mitchell reached this theory is explored. A major factor was the role of Mitchell's philosophical system conceived in his student days at Cambridge. This system appears to have become a tacit influence on his work in the sense that Polanyi understood all knowledge to be generated by an interaction between tacit and explicit knowing. Early in his life Mitchell had evolved a simple philosophy based on fluctoids, fluctids and statids which was developed in a thesis submitted for the Ph.D. at the University of Cambridge, England. This aspect of his work was rejected by the examiners and became a tacit element in his intellectual development. It is argued from his various publications that this philosophy can be traced as an underlying theme behind much of Mitchell's theoretical writing in the 50's leading, through his notion of vectorial metabolism, to the formulation and amplification of the chemiosmotic theory in the sixties. This philosophy formed the basis for Mitchell of his understanding of biological systems and gave him his unique approach to cell biology. PMID- 11859887 TI - August Weismann on germ-plasm variation. AB - August Weismann is famous for having argued against the inheritance of acquired characters. However, an analysis of his work indicates that Weismann always held that changes in external conditions, acting during development, were the necessary causes of variation in the hereditary material. For much of his career he held that acquired germ-plasm variation was inherited. An irony, which is in tension with much of the standard twentieth-century history of biology, thus exists - Weismann was not a Weismannian. I distinguish three claims regarding the germ-plasm: (1) its continuity, (2) its morphological sequestration, and (3) its variational sequestration. With respect to changes in Weismann's views on the cause of variation, I divide his career into four stages. For each stage I analyze his beliefs on the relative importance of changes in external conditions and sexual reproduction as causes of variation in the hereditary material. Weissmann believed, and Weismannism denies, that variation, heredity, and development were deeply intertwined processes. This article is part of a larger project comparing commitments regarding variation during the latter half of the nineteenth century. PMID- 11859888 TI - Essay review: improving Americans. PMID- 11859889 TI - Essay review: when evolution became conversation: Vestiges of Creation, its readers, and its respondents in Victorian Britain. PMID- 11859890 TI - Essay review: botanists sow, historians reap. PMID- 11859891 TI - Health care choices: sharing the quality message. AB - Information on health-care quality (in the form of report cards) is playing an increasingly important role in consumers' health-care decision making. In this brief we discuss how you can help your clients sort through available quality information and how you can prepare a report card using the approach developed by the Foundation for Accountability (FACCT), our guest authors this month. PMID- 11859892 TI - Allaying the threat of biological weapons. PMID- 11859893 TI - Studies of dietary fat and heart disease. PMID- 11859894 TI - The role of the PROs. AB - Peer review organizations (PROs) play a critical part in the Medicare program, yet their efforts on behalf of beneficiaries are not well-known. In this brief we explore the role of the PROs, focusing specifically on quality improvement, appeals, and beneficiary education. PMID- 11859895 TI - Building coalitions. AB - Educating people on Medicare about their benefits and rights is a task often undertaken by multiple organizations in a given community. In some areas these groups have chosen to work together as a coalition in order to pool their capabilities and resources. In this brief, we explore the topic of coalition building, focusing on specific "how-to's" and using real-life examples from the experiences of the Coalition to Monitor Medicare Managed Care (CMMMC) in Cleveland, OH. PMID- 11859896 TI - Developing culturally appropriate Medicare education materials. AB - Cultural differences are some of the most subtle, yet critical, factors affecting how people with Medicare understand their benefits, rights and choices. This brief explores the lessons we have learned from an effort to develop and test Medicare education materials suitable for audiences of varying cultural backgrounds. PMID- 11859897 TI - Clinical policy for the management and risk stratification of community-acquired pneumonia in adults in the emergency department. AB - This clinical policy represents an approach that emphasizes key clinical information to determine the severity of CAP. By using this approach, a determination of whether the patient can be treated as an outpatient or inpatient may be made. Recommendations about the utility of ancillary studies and the use of antibiotics are also given. As more of the questions are answered through controlled studies, an evidence-based approach to this problem will become increasingly important in improving the outcome of patients with CAP. PMID- 11859898 TI - Medicare preventive services. AB - Preventive services, such as mammograms and colorectal cancer screenings, can save lives. Since 1997, Medicare has had a renewed emphasis on wellness through its expanded preventive health benefits. In this brief we discuss the importance of screenings and prevention, focusing specifically on Medicare coverage of these benefits. Finally we spotlight the National Diabetes Education Program, an awareness campaign that you and your organization can be part of. PMID- 11859899 TI - Medicare ambulance service. AB - Medicare coverage of ambulance services is a topic that receives little attention in educational materials but is a major source of confusion for people with Medicare, their families and professionals. In this brief we discuss and clarify coverage issues and payment policies for ambulance services, including the potential costs for consumers. We conclude with a look at situations where appeals of denial of payment for these services may be appropriate. PMID- 11859900 TI - Increasing enrollment for the Medicare Savings Programs. AB - Financial assistance for people with Medicare with low or moderate incomes is available through the Medicare Savings Programs. Yet many people who could use these benefits are unaware that they exist or are reluctant to apply for them. This brief describes efforts that you can undertake to increase awareness about the Medicare Savings Programs and to help your clients apply for these programs. PMID- 11859901 TI - The evolution of the psyche and society. PMID- 11859902 TI - Making good better: approaches to evaluating the quality of programs and services. AB - While most agencies know that they should be evaluating the quality of the programs and services they provide, including Medicare education, often they don't know how or why they should do that. In this brief we explore the topics of quality assurance and program evaluation, including their similarities and differences, why they're important, why staff often react unfavorably to them, and how you can go about achieving quality in your programs and services. PMID- 11859903 TI - The Medicare hospice benefit. AB - November is national hospice month. Since 1983, Medicare has covered hospice services for beneficiaries with terminal illnesses. However, relatively few people know that this benefit exists. In this brief we discuss how the Medicare hospice benefit works--who is eligible, what it covers (and doesn't cover) and how you can help your clients access this benefit. PMID- 11859904 TI - The PACE model. AB - Programs of All-inclusive Care for the Elderly (PACE) have been in existence for nearly three decades but have only recently become an established part of the Medicare program. In this brief we explore the PACE model: what it is, how it works, who is enrolled and the challenges it faces in the future. PMID- 11859905 TI - Cost savings in migraine associated with less chest pain on new triptan therapy. AB - OBJECTIVES: This article constructs an economic model to estimate cost of chest pain-related care in migraine patients receiving almotriptan 12.5 mg compared with those receiving sumatriptan 50 mg. STUDY DESIGN: This population-based, retrospective cohort study used data from the MEDSTAT Marketscan database (Ann Arbor, Michigan) to quantify incidence and costs of chest-pain-related diagnoses and procedures. After a 6-month exclusion period, the study used a pre-post design, with baseline and treatment periods defined, respectively, as 5 months before and after receiving sumatriptan therapy. An economic model was constructed to estimate annual cost savings per 1,000 patients receiving almotriptan instead of sumatriptan as a function of differing rates of chest pain. Annual direct medical cost avoided was calculated for a hypothetical health plan covering 1 million lives. RESULTS: Among a cohort of 1,390 patients, the incidence of chest pain-related diagnoses increased significantly (43.6%) with sumatriptan, from 110 during the baseline period to 158 during the treatment period (P= .003). Aggregate costs for chest-pain-related diagnoses and procedures increased 33.1%, from $22,713 to $30,234. Payments for inpatient hospital services rose 10-fold; costs for primary care visits and outpatient hospital visits rose 53.1% and 14.4%, respectively. Payments for angiography increased from $0 to $462, and costs for chest radiographs and electrocardiograms increased 58.7% and 31.2%, respectively. Sumatriptan treatment was associated with a 3-fold increase in payments for services for painful respiration and other chest pain. The model predicted $11,215 in direct medical cost savings annually per 1000 patients treated with almotriptan instead of sumatriptan. Annual direct medical costs avoided for the health plan totaled $195,913. CONCLUSION: Using almotriptan instead of sumatriptan will likely reduce the cost of chest-pain-related care for patients with migraine headaches. PMID- 11859906 TI - Migraine: diagnosis, management, and new treatment options. AB - OBJECTIVE: The safety and tolerability of medications used to treat acute migraine attacks are summarized, the classification of headaches and the causes of and diagnostic criteria for migraine are reviewed, and the clinical tolerability profiles and therapeutic benefits of second-generation triptans are presented. BACKGROUND: Migraine is a paroxysmal disorder characterized by attacks of headache, nausea, vomiting, photophobia, and phonophobia. Drugs used to prevent migraine and those that effectively treat acute migraine attacks are readily available. METHODS: Mild or moderate migraines are often treated with aspirin, acetaminophen, nonsteroidal anti-inflammatory drugs, antiemetic drugs, or isometheptene. Triptans (5-HT1 receptor agonists) are used to treat moderate or severe migraine and when nonspecific medications have been ineffective. Because sumatriptan, the first triptan used, is effective but can induce adverse events, second-generation triptans (zolmitriptan, naratriptan, rizatriptan, and almotriptan) were developed to increase the benefit-to-risk ratio in migraine management. RESULTS: Important pharmacologic, pharmacokinetic, and clinical differences exist among those drugs, but the tolerability profile of the newer triptans is very good, and they provide rapid relief from headache and sustained duration of effect. CONCLUSION: Primary care physicians must manage migraine patients with treatments that demonstrate a balance between efficacy and tolerability. PMID- 11859907 TI - Almotriptan: a review of pharmacology, clinical efficacy, and tolerability. AB - OBJECTIVE: This article summarizes preclinical and clinical data for almotriptan. METHODS: Almotriptan has been evaluated in more than 3,500 acute migraine patients in phase 2 and 3 double-blind, randomized trials and in 1,500 patients in long-term open-label trials. RESULTS: Controlled clinical trials show that almotriptan 12.5 mg is significantly more effective than placebo. These results are observed across different endpoints examined, including pain relief at 2 hours, pain-free outcome at 2 hours, recurrence rate, use of escape medication, and sustained pain-free outcome. The onset of pain relief is observed as early as 30 minutes after administration. Results from a multiple-attack study show that almotriptan maintains a consistency of response across 3 attacks, and results from long-term studies confirm that patients continue to respond to almotriptan for up to 1 year. Results from 2 comparative studies show that almotriptan 12.5 mg has comparable efficacy to sumatriptan 50 or 100 mg, but almotriptan has a superior tolerability profile. Early use of almotriptan results in a higher proportion of patients achieving pain relief or complete freedom from pain. CONCLUSIONS: Because almotriptan has a tolerability profile comparable to that of placebo, it may be more acceptable for early administration. The incidence of treatment-related adverse events with almotriptan is comparable to that of placebo and significantly lower than recorded with sumatriptan. In addition, almotriptan has a low incidence of chest symptoms, an adverse event associated with triptan use. Because of its comparable efficacy and superior tolerability profile, almotriptan offers a potential improvement over existing triptans for the treatment of acute migraine. PMID- 11859908 TI - Economic comparison of oral triptans for management of acute migraine: implications for managed care. AB - Sound, informed decision making regarding which drugs to include on a formulary should be based on the best available evidence of their clinical efficacy and incidence of adverse events. Comparative drug costs and clinical effectiveness should also be considered during the formulary development process. Clinical trials traditionally evaluate efficacy and adverse events independently, whereas effectiveness in real-life conditions is defined as some combination of efficacy and side effects. When evaluating similar medications, head-to-head efficacy and effectiveness studies are preferred. For oral triptans (serotonin 5-HT(1B,1D) receptor agonists), there are many placebo-controlled trials and several active trials that compare newer oral triptans with sumatriptan; however, there have been few comparisons of triptans in head-to-head trials. Meta-analysis is an appropriate method to evaluate multiple clinical trials critically and combine the results. A recently published meta-analysis used patient-level data to assess efficacy and adverse events across multiple triptan clinical trials. In this analysis, we combined those results with medication costs to assess the overall value among oral triptans. Using this combined approach, almotriptan was found to have the greatest economic value. It delivers comparable efficacy, placebo-like tolerability, and the highest value when compared with other triptans currently marketed in the United States. PMID- 11859909 TI - Health outcomes evaluations: estimating the impact of almotriptan in managed care settings. AB - OBJECTIVE: Almotriptan, a new treatment for acute migraine attacks, may improve health outcomes while reducing expenditures on drug costs and the costs associated with unwanted adverse events. This article describes the clinical, humanistic, and economic data available for almotriptan and compares these data with published data for sumatriptan, which is considered the gold standard comparator for triptans. PATIENTS AND METHODS: Clinical trial data and published materials describing the pharmacoeconomics of sumatriptan were reviewed. RESULTS: Formulary and prescribing decision makers consider clinical, humanistic, and economic dimensions when evaluating migraine treatments. Within those dimensions, critical elements to consider include efficacy, tolerability, health-related quality of life, patient satisfaction, productivity, medication cost, and healthcare resource use. In direct comparisons, almotriptan has demonstrated efficacy, short-term health-related quality of life, and productivity results similar to sumatriptan 50 mg. Almotriptan also has an improved tolerability profile compared with sumatriptan, including a lower incidence of drug-related adverse events, a lower incidence of chest pain, and better patient satisfaction in terms of adverse events. CONCLUSIONS: Almotriptan and sumatriptan exhibit comparable efficacy for the treatment of acute migraine. The acquisition cost of almotriptan is lower than sumatriptan, and given its excellent tolerability profile, almotriptan may be preferred by patients for the treatment of migraine attacks. PMID- 11859911 TI - Characteristics of glucose metabolism in the visual cortex of amblyopes using positron-emission tomography and statistical parametric mapping. AB - BACKGROUND: The effects of amblyopia on the glucose metabolism in the visual cortex in the resting state are evaluated, the asymmetry of glucose metabolism in the ipsilateral and the contralateral occipital lobes was examined by comparing the number of hypometabolic pixels in both occipital lobes, and the correlation between this asymmetry and the results of the ophthalmologic tests was evaluated. METHODS: Eleven amblyopes (7 anisometropic and 4 strabismic) and 12 normal subjects were studied with their eyes open, but without any further visual stimulus using F-18-fluorodeoxyglucose positron-emission tomography (PET) and statistical parametric mapping. Ophthalmologic tests including stereoacuity, contrast sensitivity function, monocular optokinetic nystagmus, and visual evoked potential (VEP) were measured. RESULTS: Compared to normal subjects, glucose metabolism decreased in Brodmann area (BA) 17, BAs 18/19, both inferior temporal lobes (BAs 37 and 20), and the superior parietal lobe (BA 7) in amblyopic patients, regardless of strabismic or anisometropic amblyopia. The laterality index of the hypometabolic pixels in the occipital lobe closely correlated with the asymmetry in the latency time of VEP (r = 0.82, P <0.05). CONCLUSION: These results suggest that PET imaging of glucose metabolism can provide a functional mapping of the visual cortex in human amblyopia. PMID- 11859910 TI - Almotriptan versus sumatriptan in migraine treatment: direct medical costs of managing adverse chest symptoms. AB - OBJECTIVE: Triptans, a popular class of drugs for treatment of migraine headaches, can cause adverse events including chest symptoms. This study estimated the direct medical costs of managing chest symptoms in patients treated for acute migraine with almotriptan or sumatriptan using an economic model. METHODS: The economic model of this study combined data from a randomized clinical trial that compared almotriptan with sumatriptan and data from a practice pattern survey of physicians. The pertinent clinical evaluation period was the time immediately after administration of the first medication dose. The model was developed from the perspective of a managed care payer. RESULTS: The average direct medical cost of managing chest symptoms that appeared after the first dose of an oral triptan was $0.22 for patients treated with almotriptan and $1.64 for patients treated with sumatriptan, a difference of $1.42 per patient. CONCLUSION: Relative to sumatriptan, treatment with almotriptan is likely to reduce patient care costs associated with chest symptom adverse events. PMID- 11859912 TI - Surgical outcomes of Epibulbar dermoids. AB - PURPOSE: To evaluate results of lamellar keratoplasty in limbal dermoid. METHODS: The ocular records were reviewed of 155 consecutive eyes with solid epibulbar dermoids that underwent lamellar keratoplasty at Dr. Rajendra Prasad Centre for Ophthalmic Sciences, All India Institute of Medical Sciences from 1977 to 1998. According to the size and location, the dermoids were managed surgically either by sectoral, annular, or central lamellar keratoplasty. RESULTS: All but 16 eyes improved cosmetically; while all the patients showed reduction in astigmatism, 116 eyes improved functionally. CONCLUSION: Excision with lamellar keratoplasty appears to be an effective means of management for extensive limbal dermoid. To avoid development of amblyopia, surgery at an early age is preferred. PMID- 11859913 TI - Optic neuritis in Asian children. AB - PURPOSE: To study the clinical spectrum of pediatric optic neuritis in Korean children. METHODS: A retrospective analysis was performed on 23 patients with onset of the disease before the age of 16 years from 1995 to 2000. The degree of initial visual loss, subsequent visual recovery, magnetic resonance images, and associated diseases was reviewed. RESULTS: There was no female predilection in the study group (43%) with a mean age of 8.9 years at presentation. Involvement was bilateral in 87% of patients. A preceding febrile illness within 2 weeks of visual symptoms was reported in 39% of patients. Of the 43 involved eyes of 23 patients, disc swelling was present in 51%, and central/cecocentral scotoma in 58% of the involved eyes. With a mean follow-up of 14 months, 79% of the patients had final vision of > or =20/40. Twenty-four (96%) of 25 eyes with intravenous steroid treatment and 10 (56%) of 18 eyes without intravenous steroids showed final visual acuity > or =20/40. Only 1 patient (4%) with multifocal signal abnormalities around the periventricular white matter developed multiple sclerosis. CONCLUSION: Korean children with optic neuritis showed bilateral involvement with papillitis. The visual prognosis was relatively good and especially better in the patients receiving intravenous steroid treatment than in those without any treatment; however, this study is not controlled. Although a longer follow-up period is required, the development of multiple sclerosis is rare in Korean children with optic neuritis. PMID- 11859914 TI - Tendon surgery in Brown's syndrome. AB - PURPOSE: Longitudinal tenectomy of the reflected tendon of the superior oblique muscle resulted in full recovery in a series of cases of Brown's syndrome after surgery by one of the authors. METHODS: A survey of the records of cases of Brown's syndrome was done to assess the criteria for patients requiring surgery and to evaluate the methods used. Included in the study were 9 eyes of patients who underwent surgery over the course of 1 year. Criteria identified in cases in this study include: preoperative compulsory head tilt in primary position of gaze, severe restriction of Hess chart in the affected eye, and an enlarged field in the healthy eye; intraoperative thickening of the tendon preventing free slippage through the trochlea in the forced duction test. The surgical method consisted of trimming a swelling that was consistently found on the reflected tendon of the superior oblique muscle in all patients undergoing surgery. Enough of the swelling was removed to allow free slippage of the tendon through the trochlea, as judged by the forced duction test. Sulphamide powder soaked in delayed action steroid compound (methylprednisolone acetate) was applied to prevent postoperative adhesions. RESULTS: A normal head posture and a satisfactory upward vergence were achieved in all patients from the first days after surgery, and were maintained throughout the follow up, as shown in the pre- and postoperative photographs and Hess charts. CONCLUSIONS: A preliminary exploration of the reflected tendon of the superior oblique muscle is warranted in cases of Brown's syndrome; when a thickening of the tendon is found, its surgical trimming is advised. Our findings suggest that delayed development of the trochlea may be a possible cause of Brown's syndrome. PMID- 11859915 TI - Galactokinase deficiency: a case report. PMID- 11859916 TI - Ophthalmic diagnosis of chronic granulomatous disease. PMID- 11859917 TI - Teddy bear granuloma of the conjunctiva. PMID- 11859918 TI - Bilateral retinal involvement as a presenting sign of neurofibromatosis 2. PMID- 11859919 TI - Mechanical eye (globe) injuries in children. AB - PURPOSE: The epidemiology of mechanical eye injuries in children was studied to identify the agents of injury and their contribution to the severity of visual loss and to suggest preventive measures. METHODS: The mode, type, and severity of injury were correlated with the visual acuity 6 months after the injury in all children with mechanical eye injuries between January 1994 to January 1999. RESULTS: Of the 68 children with mechanical eye injuries, the mode of injury was host (child) related in 12 (17.65%) patients, agent related in 40 (58.82%) patients, and environment related in 16 (23.53%) patients. Mild injuries were seen in 22 (32.35%) patients, 31 (45.59%) patients had moderate injuries, and 15 (22.06%) patients had severe injuries. None of the patients with host-related injuries had a severe injury. Six (66.67%) patients with host-related injuries had a good visual outcome and none had a poor outcome. Among patients with agent related injuries, 11 (25%) had a good outcome, 14 (40%) patients had a fair outcome, and 10 (22.5%) patients had a poor outcome. Of the patients with environment-related injuries 3 (33.33%) each had good, fair, and poor visual outcomes. CONCLUSIONS: Agent and environment-related injuries had a far worse outcome than host-related injuries. This epidemiological classification directly suggests practical preventive measures that can be adopted at home or at school to reduce the incidence and severity of ocular injuries. The other predictors of the final visual outcome were the severity of the injury at presentation and the necessity for a secondary surgical procedure. PMID- 11859920 TI - Ocular myasthenia presenting as progressive external ophthalmoplegia. PMID- 11859921 TI - Management of intermittent exotropia. PMID- 11859922 TI - Nocturnal light pulses selectively induce Egr-1/NGFI-A protein in periventricular hypophysiotrophic somatostatinergic neurons. AB - Recent reports showing that circadian light cues may induce gene-specific patterns of expression in periventricular hypothalamic neurons has extended the functional correlates of light-pulse stimuli, which are conventionally restricted to circadian phase-setting actions within the suprachiasmatic nucleus (ScN). The aims of the present study were, first, to broaden these observations to the protein level using an Egr-1/NGFI-A antibody, and second to investigate the cell type specificity of induction with respect to associated hypophysiotrophic neurons. In order to co-localize Egr-1 with neuroendocrine peptides, a same species, double immunofluorescence protocol has been used. The Egr-1 antibody used for immunohistochemistry was first characterized by Western analysis, and we have shown that the C19 antisera detects a single 75 kDa protein species, which has a primarily nuclear subcellular localization. Immunohistochemical analysis has shown that Egr-1 protein is markedly induced by a 1 h nocturnal light pulse both in the body of the suprachiasmatic nucleus and in a dorsal ScN zone, which extends up into the periventricular nucleus (PeN). In contrast, induction of Egr 1 was not observed within the arcuate region of the hypothalamus. Double immunofluorescence histochemistry has shown that Egr-1 is extensively, although not exclusively, co-localized with somatostatin in PeN neurons. The selective induction of the 75 kDa protein product of the egr-1 gene in PeN somatostatinergic neurons, as contrasted with hypophysiotrophic (somatostatin and growth hormone-releasing hormone [GH-RH]) neurons of the arcuate nucleus is indicative of a cell- and stimulus-specific neuroendocrine paradigm, which may be used to address temporal characteristics of somatostatin function that determine pulsatile growth-hormone secretion. PMID- 11859923 TI - Polymorphism of the angiotensin-converting enzyme gene in patients with cerebral infarction in Koreans. AB - The relationship between cerebrovascular disease and an insertion/deletion (I/D) polymorphism in the angiotensin-converting enzyme (ACE) gene is still being debated. The frequency of the DD genotype of the ACE gene was significantly higher in subjects with than those without cerebral infarction in Japan. The aim of the present study was to assess the relationship between ACE gene polymorphism and the development of cerebral infarction in a population from Korea. We examined its possible role as a risk factor in patients with cerebral infarction. The association between ACE gene polymorphism and cerebral infarction was examined in 106 patients with cerebral infarction and 498 controls without cerebral infarction. Frequencies of the genotypes and alleles of the ACE gene were investigated. The ACE genotype was analyzed by the polymerase chain reaction (PCR). The frequency of D allele was 37.7% in patients and 39.1% in controls (chi2 = 0.128, p = 0.720). The frequencies of the genotypes of the ACE gene were II: 39.6%, ID: 45.3%, and DD: 15.1% in patients, and II: 37.1%, ID: 47.6%, and DD: 15.3% in controls (chi2 = 0.127, p = 0.721). There was no significant difference in the frequency of the DD genotype of the ACE gene, and we did not find any association between ACE polymorphism and cerebral infarction. These results indicate that ACE polymorphism is not a risk factor for the development of cerebral infarction in a Korean population. PMID- 11859924 TI - Roles of BCL-2 and caspase 3 in the adenosine A3 receptor-induced apoptosis. AB - Selective A3 adenosine receptor agonists have been shown to induce apoptosis in a variety of cell types. In this study we examined the effects of adenosine receptor agonists selective for A1, A2A, or A3 receptors on the induction of apoptosis in primary cultures of rat astrocytes and in C6 glial cells. Treatment of the cells with the A3 receptor agonist Cl-IB-MECA (10 microM) induced apoptosis in both cell types. The effects of Cl-IB-MECA were partially antagonized by the A3 receptor-selective antagonist MRS 1191. In contrast, the A1 and A2A receptor agonists, CPA and CGS 21680, respectively, did not have significant effects on apoptosis in these cells. Cl-IB-MECA reduced the expression of endogenous Bcl-2, whereas it did not affect the expression of Bax. Overexpression of Bcl-2 in C6 cells abrogated the induction of apoptosis induced by the A3 agonist. Cl-IB-MECA also induced an increase in caspase 3 activity and caspase inhibitors decreased the apoptosis induced by the A3 agonist. These findings suggest that intense activation of the A3 receptor is pro-apoptotic in glial cells via bcl2 and caspase-3 dependent pathways. PMID- 11859925 TI - A double cysteine trkA mutant exhibiting reduced NGF binding and delayed Erk signaling. AB - The NGF receptor trkA is a tyrosine kinase receptor comprising an extracellular domain with a ligand-binding site, a transmembrane-spanning domain (TMD), and an intracellular domain composed of a juxtamembrane region (JMR), a tyrosine kinase domain, and a short carboxy-terminal tail. Nerve growth factor (NGF) binds and activates this receptor, leading to phosphorylation of signaling substrates involved in neuronal proliferation, differentiation, and survival. Human trkA contains one cysteine residue in the TMD (C423) and another, separated by 12 residues, in the JMR (C436). We hypothesized that the removal of one or both of the cysteines would affect NGF-induced signaling of the trkA receptor. Here we show that NGF induces rapid receptor autophosphorylation in a wild-type, trkA expressing clone (WT11), in a single cysteine trkA mutants (C423T or C436A), but lower autophosphorylation activity in a double-cysteine trkA mutant (C423T/C436A). WT11 and SM cells had similar binding affinity, but that of DM cells was lower, according to the NGF radioreceptor assay. NGF-induced Erk phosphorylation was rapid in WT11 and C423T cells, but delayed in C436A and C423T/C436A cells. NGF induced [3H]thymidine incorporation into WT11 and SM cells, but had no effect on DM cells. However, basic fibroblast growth factor (bFGF) induced rapid phosphorylation of Erk1/2, and [3H]thymidine incorporation in NIH3T3, WT11, single mutant (SM), and double mutant (DM) cells, suggesting that the impaired NGF-induced Erk phosphorylation and thymidine incorporation observed in DM cells are due to the double-cysteine mutations in the trkA receptor. Cumulatively, our findings support a model in which Cys436 of the trkA is responsible for the rapid transfer of the transmembrane occupancy signal to the SHC adaptor protein for activation of the Ras-Erk pathway and DNA synthesis. PMID- 11859926 TI - Tetanus toxin modulates serotonin transport in rat-brain neuronal cultures. AB - As has been previously described, tetanus toxin (TeTx) and its H(C) fragment inhibit the sodium-dependent 5-hydroxytryptamine (5-HT) uptake in rat-brain synaptosomes, probably through a kinase mechanism affecting the 5-HT transporter. Now, the inhibition of 5-HT uptake in neurons in primary culture by TeTx in a dose-dependent manner is described in this work. This effect is also produced by the nontoxic C-terminal fragment of the TeTx heavy chain (H(C)-fragment), indicating that 5-HT uptake inhibition is a consequence of the toxin binding to the plasmatic membrane and not to its catalytic activity. This conclusion is supported by the fact that the 5-HT accumulation was not inhibited by the light chain of TeTx or the toxoid, and was even potentiated by botulinum neurotoxin A. These results correlate with the activation of phosphoinositide-phospholipase C activity in the cultures used in this study, this activity only being enhanced by TeTx and by its Hc-fragment. On the other hand, the use of tyrosine phosphorylation modulators indicates that both Na3VO4 and basic fibroblast growth factor (bFGF) produce an enhancement of 5-HT uptake in this system, which is also sensitive to TeTx inhibition. On the other hand, genistein alone is able to reduce the 5-HT transport in cultured neurons, and this effect did not appear to be additive to that elicited by TeTx. This result suggests that TeTx and genistein may share some events in their respective mechanisms of action. Furthermore, the incubation at different concentrations of 12-O tetradecanoylphorbol 13-acetate (TPA) confirms the involvement of protein kinase C (PKC) in 5-HT transport modulation in rat-brain neuronal primary cultures. In summary, we shall demonstrate in this work that TeTx induces, through its Hc fragment, an inhibition of both basal and stimulated serotonin uptakes in primary neuronal cultures, in parallel to the activation of phosphoinositide phospholipase C activity and PKC activation. PMID- 11859928 TI - Characterization of a G protein coupling "YL" motif of the human VPAC1 receptor, equivalent to the first two amino acids in the "DRY" motif of the rhodopsin family. AB - The conserved residues Y239 and L240 of human VPAC1 receptor are predicted to be at the same location as the asparagine and arginine in the "DRY" motif in the Rhodopsin family of G protein-coupled receptors. By comparing vasoactive intestinal peptide (VIP) binding with or without the presence of GTP-gamma-S, it was found that the deltadelta G(o) for the endogenous G-protein coupling was 1.5 kJ/mol, 0.95 kJ/mol, and 3.4 kJ/mol for theY239A, L240A, and wild-type receptor, respectively. VIP-induced cAMP production in whole cells support the results of the binding studies, as Y239A had a moderate and L240A a pronounced impaired ability to produce cAMP. The mutants had a minor influence on the intrinsic "low affinity to high affinity equilibrium," suggesting that the dominating effect of these mutants is a perturbation of the G protein-binding site. Thus, the highly diverged chemical properties of the hydrophobic "YL" motif and charged "DR(Y)" motif could be a crucial difference between the Secretin Receptor Family and the Rhodopsin Family with respect to receptor activation and G-protein coupling. PMID- 11859927 TI - Expression and fine mapping of murine vasoactive intestinal peptide receptor 1. AB - Vasoactive intestinal peptide (VIP) plays multiple roles in the nervous, endocrine, and immune systems as a neurotransmitter, a hormone, and a cytokine. VIP is widely distributed in neurons of the central and peripheral nervous systems (CNS/PNS), and recently has been found to be an important neuroprotective agent. VIP actions are mediated through specific G protein-coupled receptors. We have cloned the cDNA of VIP receptor subtype 1 (VIPR1 or VPAC1) and have demonstrated the quantitative expression profile in mice. Fluorometric real-time reverse transcription-polymerase chain reaction (RT-PCR) analysis demonstrated that VPAC1 is expressed in all tissues examined. Expression was highest in the small intestine and colon followed by the liver and brain. The high level of VPAC1 expression in forebrain and cerebellum suggests that VPAC1 may mediate the neuroprotective effect of VIP. We have refined the chromosomal localization of the mouse, rat, and human VPAC1 genes. This fine mapping of the VPAC1 gene extends the respective regions of synteny between the distal region of mouse chromosome 9, rat chromosome 8q32, and human chromosome 3p21.33-p21.31. Thus, VPAC, constitutes a functional-positional candidate for the tumor-suppressor function mapped to human 3p22-p21 where loss-of-heterozygosity is observed in small-cell lung carcinoma (SCLC) cell lines and primary tumors. Availability of the cDNA sequences for mouse VPAC1 will facilitate the generation of VPAC1 null mutant animals. Such studies will ultimately enhance our understanding of the role of VIP in the nervous system. PMID- 11859929 TI - A vasoactive intestinal peptide antagonist inhibits the growth of glioblastoma cells. AB - The effects of a vasoactive intestinal peptide (VIP) receptor antagonist (VIPhyb) on human glioblastoma cells were characterized. Pituitary adenylate cyclase activating polypeptide (125I-PACAP-27) bound with high affinity to U87, U118, and U373 cells. Specific 125I-PACAP-27 binding to U87 cells was inhibited, with high affinity, by PACAP but not VIP or VIPhyb (IC50 = 10, 1500, and 500 nM, respectively). By reverse transcriptase-polymerase chain reaction (RT-PCR), a major 305 bp band was observed indicative of PAC1 receptors. PACAP-27 caused cAMP elevation and the increase in cAMP caused by PACAP-27, was inhibited by the VIPhyb. Also, PACAP-27 caused cytosolic Ca2+ elevation in Fura-2AM loaded U87 cells and the VIPhyb inhibited this increase. Using the MTT growth assay, the VIPhyb was shown to inhibit glioblastoma growth in a concentration-dependent manner. Using a clonogenic assay in vitro, 10 microM VIPhyb significantly inhibited proliferation of U87, U118, and U373 cells. In vivo, 0.4 microg/kg VIPhyb inhibited U87 xenograft proliferation in nude mice. These results suggest that the VIPhyb antagonizes PAC1 receptors on glioblastoma cells and inhibits their proliferation. PMID- 11859930 TI - Increased gene expression of interleukin-1alpha and interleukin-6 in rat primary glial cells induced by beta-amyloid fragment. AB - One of the pathological hallmarks of Alzheimer's disease (AD) is the presence of amyloid plaques. The main constituent of the amyloid plaques is the amyloid beta peptide (A beta) shown to activate glial cells in vitro. A growing body of evidence suggests that these cells contribute to neurotoxicity through production of inflammatory cytokines, chemokines, and neurotoxic substances, such as reactive oxygen species (ROS). In this study, mRNA levels of the inflammatory cytokines interleukin (IL)-1alpha and beta, and IL-6 were analysed by reverse transcriptase-polymerase chain reaction (RT-PCR) in rat primary mixed glial cells after treatment with A beta(25-35), a biologically active fragment of A beta peptide with neurotoxic properties. Clear morphological changes of the astrocytes, as well as proliferation and clustering of microglial cells were observed by light and immunofluorescence microscopy after 24 h treatment. Significant increases in IL-1alpha and IL-6 mRNA levels were detected after 24 and 72 h, whereas significantly increased levels of IL-1beta mRNA could only be detected after 4 h treatment. The most pronounced effect was seen on IL-6 mRNA expression, which increased approx two- to threefold after treatment. In addition, increased secretion of IL-6 was detected after 96 h exposure. Recently, association of IL-1alpha and IL-6 gene polymorphism with AD was reported, suggesting that these cytokines may play an important role in the development of the disease. The increased mRNA levels of IL-1alpha and IL-6 in parallel with the morphological changes in the mixed glial-cell cultures support that these cytokines may be involved in A beta-induced gliosis and in the pathogenesis of AD. PMID- 11859931 TI - Differential expression of carnosine, homocarnosine and N-acetyl-L-histidine hydrolytic activities in cultured rat macroglial cells. AB - N-acetyl-L-histidine (NAH) and N-acetyl-L-aspartate (NAA) are representatives of two series of substances that are synthesized by neurons and other cells in the vertebrate central nervous system (CNS). Histidine containing homologs of NAH are beta-alanyl-L-histidine or carnosine (Carn) and gamma-aminobutyrl-L-histidine or homocarnosine (Hcarn). A homolog of NAA is N-acetylaspartylglutamate (NAAG). These substances belong to a unique group of osmolytes in that they are synthesized in cells that may not to be able to hydrolyze them, and are released in a regulated fashion to a second compartment where they can be rapidly hydrolyzed. In this investigation, the catabolic activities for NAH, Carn, and Hcarn in cultured macroglial cells and neurons have been measured, and the second compartment for NAH and Hcarn has been identified only with astrocytes. In addition, oligodendrocytes can only hydrolyze Carn, although Carn can also be hydrolyzed by astrocytes. Thus, astrocytes express hydrolytic activity against all three substrates, but oligodendrocytes can only act on Carn. The cellular separation of these hydrolytic enzyme activities, and the possible nature of the enzymes involved are discussed. PMID- 11859932 TI - Selective loss of KGDHC-enriched neurons in Alzheimer temporal cortex: does mitochondrial variation contribute to selective vulnerability? AB - THE RESEARCH OBJECTIVE: of this study was to test whether variation in mitochondrial composition is associated with "selective vulnerability" in Alzheimer brain. The term "selective vulnerability" refers to the loss of relatively vulnerable brain cells and the sparing of relatively resistant brain cells in disorders in which a genetic defect or environmental agent acts on both types of cells. The mechanisms underlying selective vulnerability are largely unknown, but mitochondria may be involved; the composition of mitochondria varies among different types of neurons, and mitochondria have an important role in cell death. Alzheimer's Disease (AD) is one of a number of neurodegenerative disorders in which both selective vulnerability and abnormalities of mitochondria occur. METHODS: We examined by immunohistochemistry the cellular distribution of a mitochondrial constituent (the alpha-ketoglutarate dehydrogenase complex, KGDHC) known to be deficient in AD, in relation to the known selective vulnerability of neurons in areas 21 and 22 of the temporal lobe in this neurodegenerative disorder. RESULTS: In normal human brain, cortical layers III and V contain neurons intensely immunoreactive for KGDHC, compared to other cells in these areas. The KGDHC-enriched cells are lost in AD (p < 0.001). In layer III, the loss of KGDHC-enriched cells is proportional to total loss of neurons, as determined by immunoreactivity to neuron specific enolase (NSE). In layer V, a higher proportion of the KGDHC-enriched neurons are lost than of other (NSE positive) neurons (p < 0.001). SIGNIFICANCE: Variations in mitochondrial composition may be one of the factors determining which cells die first when different types of cells are exposed to the same stress. PMID- 11859934 TI - Towards an understanding of biological role of colostrinin peptides. AB - The aim of this study was to elucidate the structure and possible function of colostrinin, also known as a proline rich polypeptide (PRP). The molecular weight of colostrinin was originally determined by gel filtration to be 17,200 daltons. In the presence of guanidinum chloride, however, the molecular weight was found to be about 6,000 daltons. Further studies utilizing high-performance liquid chromatography (HPLC) and mass spectroscopy revealed that colostrinin is a complex consisting of many low molecular-weight polypeptides. A total of 32 peptides were isolated from the original colostrinin preparation by HPLC and subjected to the N-terminal sequence analysis. The results of sequence analysis revealed significant homology of the peptides to three protein precursors: annexin, beta-casein, and a hypothetical beta-casein homolog. In addition, the sequence of several peptides showed no significant homology to any specific protein in the current GenBank database. The synthetic peptides of various lengths representing the N-terminal sequence of the colostrinin peptides were made to study some biological effects. Here we report that colostrinin and some of its component peptides are potent inducers of leukocyte proliferation and of certain cytokines. Also, a series of monospecific antibodies were produced in rabbits against the synthetic peptides. The antibodies were used to study the kinetic of antigen reduction in colostrum and mature milk following lambing. A threefold decrease was common for most antigens studied over the period of 72 h. Based on the results of these studies we postulate that colostrinin represents a diverse group of peptides produced in the mammary gland of mammals for the development of the optimal physiologic responses in offspring. Also, it is hoped that the beneficial use of colostrinin in Alzheimer's Disease (AD), recently reported elsewhere, will revive interest in its clinical application for treatment and/or prophylaxis of many age-related disorders. PMID- 11859933 TI - Gangliosides inhibit the release of interleukin-1beta in amyloid beta-protein treated human monocytic cells. AB - Amyloid-beta protein (A beta) is known to induce microglial activation with concomitant release of cytokines. Gangliosides have documented neuritogenic and neurotrophic properties. We determined the effects of A beta on the release of interleukin-1beta (IL-1beta) from the human monocytic cell line, THP-1 cells. A beta 1-42 significantly induced the release of IL-1beta from the cells. A beta 1 40, A beta 40-1, A beta 1-38, and A beta precursor protein (beta-APP) analogs also released a small amount of IL-1beta. A beta 1-42-activated cells demonstrated approx an 18-fold higher IL-1beta release than that for control cells or A beta 1-40 (soluble; S) treated cells. The release of IL-1beta from A beta 1-42-activated cells was significantly inhibited (33-48% of activated cells; p < 0.05 for the control value) by addition of gangliosides, suggesting that gangliosides inhibit the continuous cycle of the IL-1beta production in THP-1 cells. PMID- 11859935 TI - Inhibition of heat shock-induced apoptosis by peppermint oil in astrocytes. AB - Exposure to environmental stresses and toxins is linked to the pathogenesis of neuropsychiatric disorders. Astrocytes, the most abundant glial-cell type in the brain, are considered to have physiological and pathological roles in neuronal activities. We have investigated whether peppermint oil inhibits heat shock induced apoptosis of astrocytes. We found that peppermint oil inhibits the heat shock-induced apoptosis in both human astrocyte CCF-STTG1 cells and rat astrocytes. Pretreatment of the cells with peppermint oil inhibited the heat shock-induced DNA fragmentation and condensation of nuclear chromatin. Peppermint oil also inhibited the caspase-3 activation and poly-ADP-ribose polymerase fragmentation in CCF-STTG1 cells. These results suggest that peppermint oil may modulate the apoptosis of astrocytes via the activation of the caspase-3. PMID- 11859937 TI - Does antisense make sense in dermatology? AB - The concept of antisense technology is elegant but misleadingly simple. Short oligodeoxynucleotides (ODNs) complementary to a target messenger RNA form DNA-RNA hybrids by Watson-Crick base pairing rules, and interfere with expression of the encoded protein. The potential sequence specificity of antisense ODNs makes them attractive as molecular drugs in the treatment of human diseases. The skin is readily accessible and, in theory, is therefore suitable for application of antisense ODNs. Targeted and selective inhibition of keratinocyte gene expression in human epidermis could be an efficient and safe pharmacological approach in a number of skin diseases. Based on recent studies from our group and others, in this review we present our view on the usefulness of antisense ODN technology in skin for the modulation of gene expression related to skin diseases. It has become clear from these studies that practising antisense technology requires careful experimentation and critical data interpretation. Although the antisense technique was applied with success in some skin model systems, we feel that the technology is still in its infancy. The basic questions have been answered, but there are still many more that need to be addressed. PMID- 11859936 TI - Identification of maze learning-associated genes in rat hippocampus by cDNA microarray. AB - Long-term memory formation requires de novo RNA and protein synthesis. To assess gene-expression changes associated with learning and memory processes, we used cDNA microarray to analyze hippocampal gene expression in male Fischer-344 rats following training in a multiunit T-maze. Here, we report the identification of 28 clones (18 known genes and 10 ESTs) for which expression increased after the maze learning. Some of the known genes appear to be involved in Ca2+ signaling, Ras activation, kinase cascades, and extracellular matrix (ECM) function, which may regulate neural transmission, synaptic plasticity, and neurogenesis. The gene expression profile presented here provides the groundwork for future, more focused research to elucidate the contribution of these genes in learning and memory processes. PMID- 11859938 TI - Efficacy of a new class of perfluoropolyethers in the prevention of irritant contact dermatitis. AB - Perfluoropolyethers (Fomblin HC products) are chemical non-reactive polymers with special physico-chemical properties that recently showed promise as protective preparations in the prevention of irritant contact dermatitis. We evaluated the efficacy of a new class of perfluoropolyethers (perfluoropolyether phosphate, Fomblin HC/P2) in the prevention of experimentally induced cumulative irritant contact dermatitis if applied prior to irritation. A panel of 20 healthy volunteers was tested with a repetitive irritation test using 4 standard irritants (sodium lauryl sulphate of highest purity, sodium hydroxide, lactic acid and toluene) in a randomized double-blind study. Application sites were assessed clinically and by the use of bioengineering techniques (transepidermal water loss and chromametry). Three gel preparations each containing 5% perfluoropolyether phosphate showed significant efficacy against irritation due to sodium lauryl sulphate and sodium hydroxide, while one test preparation containing 2% showed inferior benefit, indicating a dose-related effect. Preparations containing perfluoropolyether phosphates can be recommended for workplaces with water-soluble irritants. Further studies under real workplace conditions are indicated. PMID- 11859939 TI - Response to thermal stimuli in skin pretreated with sodium lauryl sulfate. AB - Skin irritation is mostly a multifactorial process. Competitive effects of different chemical irritants are well known. This study investigates the influence of a thermal stimulus on skin pre-irritated with sodium lauryl sulfate (SLS). Seventy-seven volunteers were patch-tested with SLS 0.25% and 0.5% for 48 h. Water served as control. Skin reaction was evaluated by measurement of transepidermal water loss, skin blood flow and skin color. After measurement, a thermal stimulus was applied on the test area. The increase in skin blood flow was measured. There was a significant correlation between the degree of irritation and the increase in skin blood flow after thermal stimulus. Pre irritated skin reacted to thermal stimulus with a shorter and sharper increase in skin blood flow. This increase was dependent on the SLS concentration. Hence, the thermally stimulated blood flow may be a model of non-chemical irritation and seems to be a relevant co-factor in the pathogenesis of irritant dermatitis. PMID- 11859940 TI - Structural analogues inhibit the sensitizing capacity of carvone. AB - The aim of the study was to investigate the effect of non-allergenic structural analogues on the sensitizing potential of carvone, a fragrance allergen. The possibility that one molecule might inhibit the allergenic activity of another molecule has been debated for 25 years. The Research Institute for Fragrance Materials states that the sensitizing activity from certain fragrance aldehydes is "quenched" by the addition of other specific chemicals. However, other studies do not confirm the results, although several attempts have been made. We used a guinea pig method designed to study the sensitizing capacity of fragrance allergens. Induction was performed with either carvone alone or with a mixture of carvone and one of two analogues. A significant difference in the response rates (p < 0.001) was observed between the animals induced with carvone alone and those induced with any of the mixtures. Our investigation shows that by using selected molecules it is possible to significantly reduce the sensitizing effect of a fragrance allergen. PMID- 11859942 TI - Differential proteinase expression by Pseudomonas aeruginosa derived from chronic leg ulcers. AB - Pseudomonas aeruginosa colonizes 20-30% of all venous leg ulcers. Hypothetically, P. aeraginosa could release proteases and cytotoxic substances in the environment of chronic ulcers, thus negatively affecting the wound-healing activity in this patient group. Here we show that P. aeruginosa isolates from leg ulcers exhibit a highly variable expression of the proteinases elastase and alkaline proteinase. We propose that bacterial phenotype should be taken into account in future studies on the clinical outcome of leg ulcers colonized by P. aeruginosa. PMID- 11859941 TI - Tandem application of sodium lauryl sulfate and n-propanol does not lead to enhancement of cumulative skin irritation. AB - Irritant contact dermatitis has a broad spectrum of clinical features and is a leading cause of occupational disease worldwide. It has been shown previously that a combination of chemically different irritants may cause an additive effect compared to single application of these substances. In this study, tandem application of sodium lauryl sulfate and n-propanol was investigated in 20 human volunteers using non-invasive bioengineering methods, such as measurement of transepidermal water loss and chromametry. N-propanol did not enhance cumulative skin irritation when used with sodium lauryl sulfate, as has been reported for toluene. As n-propanol is the active ingredient in many disinfectants, this is of particular interest regarding occupational skin irritation in health care workers. PMID- 11859943 TI - Increased psychiatric morbidity in female outpatients with skin lesions on visible parts of the body. AB - Psychiatric disorders are frequent among patients with skin diseases. We aimed at identifying factors associated with psychiatric morbidity in dermatological outpatients. All adults attending the outpatient clinics of a dermatological hospital on predetermined days were given the 12-item General Health Questionnaire. The dermatologists indicated the diagnosis and location of skin lesions and rated the disease severity. A total of 1389 patients were asked to take part in the study. Of the 722 who accepted, 389 had a complete set of data and were included in the analysis. To verify the representativeness of our sample, we used the administrative registries to compare participants with the total population of patients who attended the clinics during the same period, and we examined the distribution of missing data. There was a tendency towards a younger age in the sample studied, but no evidence of substantial selection bias. The prevalence of psychiatric morbidity was 20.6%. We found higher probability of psychiatric disorders in women, controlling for age, clinical severity and localization of lesions. In women, but not in men, the prevalence of psychiatric morbidity was higher in patients with lesions on the face or hands. Given that the identification and appropriate management of psychiatric morbidity are important, it seems that the dermatologist should be particularly alert to the possibility of a concurrent psychiatric disorder in women with lesions on the face or hands. PMID- 11859944 TI - Occupational non-melanoma skin cancer. AB - Non-melanoma skin cancer is historically known to be associated with certain professions. Reporting is mandatory in Denmark when occupational exposure is suspected. In a retrospective register-based study of all cases of suspected occupational non-melanoma skin cancer reported to the Directorate of National Labour Inspection and the National Board of Industrial Injuries in Denmark in the period January 1, 1984 to December 31, 1994, we assessed the extent to which occupational exposures today are of importance in the occurrence of non-melanoma skin cancer. A total of 74 individuals (11 women and 63 men) aged 32-82 years (median 58 years) had been reported. Of these, 15 cases (20%) were approved as being occupational, 37 (50%) were rejected and 22 (30%) were either shelved or could not be further clarified. Most commonly approved were exposures such as asphalt, tar, and the like, and ionizing radiation, and localization on the arms or multiple tumours. Unexpected occupational exposure could not be identified but continued reporting is recommended in order to follow this in the future. PMID- 11859945 TI - Serum IgE reactivity to Malassezia furfur extract and recombinant M. furfur allergens in patients with atopic dermatitis. AB - IgE reactivity to the opportunistic yeast Malassezia furfur can be found in patients with atopic dermatitis (AD). We have previously cloned and expressed 6 recombinant allergens (rMal f 1, rMal f 5-9) from M. furfur. In the present study, we used ImmunoCAP to investigate whether these rMal f allergens can be useful in the diagnosis of M. furfur-associated AD compared with the M. furfur extract. A total of 156 adult patients with a clinical diagnosis of AD participated in the study. Sixty-four percent had increased total serum IgE levels, 79% had specific IgE antibodies to common inhalant allergens and 47% had IgE antibodies to M. furfur extract. IgE antibodies to any of the rMal f allergens were detected among 86 (55%) of the patients, 14 (16%) of whom did not react to the M. furfur extract. Any individual rMal f allergen detected between 32% and 89% of the patients ImmunoCAP-positive to the M. furfur extract, with the highest sensitivity for rMal f 9. Therefore, a couple of individual rMal f allergens can improve the diagnosis of M. furfur-associated IgE allergies in patients with AD. PMID- 11859946 TI - Effects of short-term oral acitretin therapy on peripheral nerve function: a prospective neurological and neurophysiological study. AB - The aim of the present prospective study was to substantiate possible side effects of short-term oral acitretin therapy (1 mg/kg/day) on peripheral nerve function of 13 patients with severe keratinization disorders. Clinical neurological examination before and 1 and 3 months after onset of treatment was unremarkable in all patients; however, a significant alteration of one or more neurophysiological, predominantly sensory, parameters was demonstrated in 3 out of 13 patients (23%) after 1 month and in 9 out of 13 (69%) after 3 months of oral acitretin therapy. These findings indicate that oral acitretin might be capable of causing a dysfunction of predominantly sensory nerve fibres in some individuals. Although in the investigated patients this dysfunction remained subclinical, it seems reasonable to suggest that neurological and neurophysiological evaluation of peripheral nerves should be added to the list of investigations that are routinely performed in patients receiving oral acitretin. PMID- 11859947 TI - Gemcitabine-induced acute lipodermatosclerosis-like reaction. AB - Gemcitabine is considered to be a well-tolerated and safe cytostatic drug because of the relative lack of side effects. Cutaneous reactions due to gemcitabine treatment, including alopecia and maculopapular eruption, are well known. We report one patient with squamous cell carcinoma of the lung who developed a gemcitabine-induced cutaneous reaction mimicking acute lipodermatosclerosis. This case has never been reported in the literature before. PMID- 11859948 TI - Epidermolysis bullosa acquisita: correlation of IgE levels with disease activity under successful betamethasone/dapsone combination therapy. PMID- 11859949 TI - Addition of pentoxifylline could reduce the side effects of fumaric acid esters in the treatment of psoriasis. PMID- 11859950 TI - Videodermatoscopic approach to porokeratosis of Mibelli: a useful tool for the diagnosis. PMID- 11859951 TI - Ultraviolet A sunbed used for the treatment of scleroderma. PMID- 11859952 TI - Sensitivity to rubber chemicals and latex among hemodialysis patients. PMID- 11859954 TI - Constitutional pompholyx eczema complicated by secondary lymphoedema. PMID- 11859953 TI - Granulocyte colony-stimulating factor (G-CSF) induced disseminated erythema with CD68 positive histiocytes. PMID- 11859955 TI - Narrow-band UVB treatment in atopic dermatitis. PMID- 11859956 TI - Acrokeratosis paraneoplastica (Bazex's syndrome): unusual association with a peripheral T-cell lymphoma. PMID- 11859957 TI - Photosensitivity from colophony in a case of chronic actinic dermatitis associated with contact allergy from colophony. PMID- 11859958 TI - Scabies on the Web? PMID- 11859959 TI - Kerion due to Trichophyton mentagrophytes: responsiveness to fluconazole versus terbinafine in a child. PMID- 11859960 TI - Extensive calcinosis cutis in association with systemic lupus erythematosus. PMID- 11859961 TI - A long-lasting verrucous plaque on the hand revealed as tuberculosis cutis. PMID- 11859962 TI - Generalized bullous fixed drug eruption after influenza vaccination, simulating bullous pemphigoid. PMID- 11859963 TI - Guest editor's introduction to a special issue: barriers to providing effective mental health services to racial and ethnic minorities in the United States. PMID- 11859964 TI - Barriers to effective mental health services for African Americans. AB - Many African Americans--especially the most marginal--suffer from mental health problems and would benefit from timely access to appropriate forms of care. However, few seek treatment from outpatient providers in the specialty mental health sector and those who do are at risk of dropping out. African Americans visit providers in the general medical sector, although they use another hypothesized alternative to specialty care, voluntary support networks, less than other groups. These help-seeking tendencies may reflect characteristic coping styles and stigma, as well as a lack of resources and opportunities for treatment. More should be learned about differences in need according to location, social standing, and cultural orientation so as to identify treatments and programs that are especially beneficial to African Americans. PMID- 11859965 TI - Priority issues in Latino mental health services research. AB - This paper identifies issues and trends affecting the quality and comprehensiveness of Latino mental health research and services. These issues include current patterns of need and services use, rapid expansion of the Latino population, extraordinary rates of uninsured, social and language barriers to care, transformation in treatment science and technology, and the sheer complexity and rapid changes in the delivery system. Progress in the field requires coordination and investments from both public and private sectors. Scientific journals should provide assistance for creating a high quality knowledge base and rapidly disseminating this information to students, practitioners, and policy makers. Vigorous activity is needed to (1) augment the supply of people entering the "pipeline" for researcher and practitioner training, and (2) support research in priority areas such as outcome studies for diverse treatments and different sectors of care, cultural competence, treatment models for youth and aging populations, quality of care, and barriers to mental health care. PMID- 11859966 TI - Barriers to providing effective mental health services to Asian Americans. AB - Using the research framework recommended by L. Rogler, R. Malgady, and D. Rodriguez (1989), the current paper examines the barriers to providing effective mental health services to Asian Americans. Beginning with the recognition that Asian Americans consists of numerous heterogeneous subgroups, the issue of the stereotype of Asian Americans as the "model minority" was also discussed. The primary focus of the paper is on Stages 2, 3, and 4 within the Rogler et al. (1989) model and the identification and discussion of cultural factors that hinder the delivery of mental health services to Asian Americans. The paper is therefore organized into these three sections: (a) help-seeking or mental health service utilization, (b) evaluation of mental health problems, and (c) psychotherapeutic services. In each of the sections, not only are the barriers to delivery of effective mental health services discussed but so are the research and methodological problems as well as some directions for future research. This critical review of the literature has been prepared with the goal of serving as a "blueprint" for us to pursue rigorous but relevant research to identify and reduce these cultural barriers to providing effective mental health services to Asian Americans. PMID- 11859967 TI - Barriers to providing effective mental health services to American Indians. AB - Like most indigenous populations throughout the world who have undergone innumerable cultural changes, the mental health care needs of American Indians are great. Some surveys conducted by the Indian Health Service show high rates of suicide, mortality, depression and substance abuse. Little is known about effective mental health care among American Indians due, in part, to the lack of culturally appropriate models of mental health in American Indians. This article presents a cultural framework in order to understand the mental health care needs of American Indians and discusses barriers to providing effective mental health services to American Indians. PMID- 11859968 TI - Macrophage/microglial cell subpopulations in glioblastoma multiforme relapses are differentially altered by radiochemotherapy. AB - Following surgical removal of glioblastoma multiforme (GBM), radiochemotherapy impedes neoplastic outgrowth and relapse formation. Macrophages/microglial cells are believed to be potent mediators of the host defense system in GBM. However, little is known about their alteration by postsurgical therapies. We have now analyzed expression of LCA (leucocyte common antigen), CD68 (phagocytic cells), HLA-DR, -DP, -DQ (MHC class II), MRP-8 (myeloid-related protein, S100A8), MRP-14 (S100A9), LCF (lymphocyte chemoattractant factor, IL-16) and NOS II (inducible nitric oxide synthase) in macrophages/microglial cells in 39 GBM relapses and their matched primary tumors. Following surgery of the primary tumors, 15 patients received irradiation and chemotherapy, 17 irradiation and 7 no treatment. In irradiated relapses, we observed significantly more macrophages/microglial cells expressing MRP-14 compared to untreated GBM relapses. Furthermore, we observed a significant increase of CD68 expressing macrophages/microglial cells in patients without postsurgical treatment, but not in those with radiochemotherapy. In conclusion, our findings suggest that radiochemotherapy alters the number of MRP-14 expressing cells. The lacking increase of CD68 expressing cells in patients with radiochemotherapy suggests depletion of this cell type by postsurgical therapy. PMID- 11859970 TI - Analysis of p53 tumor suppressor gene in families with multiple glioma patients. AB - The high incidence of gliomas in Li-Fraumeni families and the high frequency of somatic p53 mutations in sporadic glial tumors have raised the possibility that germline p53 mutations could play an important role in familial aggregation of gliomas. In the present study, 18 families with two or more gliomas were screened for germline p53 mutation. The families were identified through questionnaires sent to 369 consecutive glioma patients operated at Tampere University Hospital during 1983-1994. In these families, a family history of cancer was verified through the Finnish Cancer Registry. Interestingly, the questionnaires reveled only 15 of 57 cancers (index gliomas excluded) retrieved through the Cancer Registry. None of the 18 families fufilled the criteria for classic Li-Fraumeni syndrome. Immunostaining analysis of p53 protein accumulation suggested that alterations of the p53 gene are as common in familial as in sporadic gliomas. Sequencing analysis of exons 4-10 of the p53 gene revealed no germline mutations in any of the 18 families. Thus, although occasional glioma families carrying germline p53 mutations have been identified in earlier studies, systematic evaluation of familial glioma patients suggests that the p53 gene is not a common susceptibility gene in case of familial gliomas. The p53 tumor suppressor gene seems to have a similar role in the tumorigenesis of most familial and sporadic gliomas. PMID- 11859971 TI - Metastatic seeding of the stereotactic biopsy tract in glioblastoma multiforme: case report and review of the literature. AB - OBJECTIVE AND IMPORTANCE: The first case was recently reported of tumor seeding by glioblastoma multiforme (GBM) after stereotactic biopsy. This occurred despite radiosurgical treatment of the lesion post-biopsy. We report the first case of metastatic seeding along the needle biopsy tract of a GBM in which the tract was within the treatment field of subsequent fractionated radiation therapy. CLINICAL PRESENTATION: A 56-year-old man presented with left-sided focal motor seizures. An MRI showed an enhancing right cingulate gyrus lesion. INTERVENTION: A stereotactic biopsy of the lesion disclosed GBM. Radiation therapy was begun 25 days after biopsy and was completed 39 days thereafter. The biopsy tract received a minimum of 60 Gy. Subsequent magnetic resonance scanning showed the lesion to have doubled in size and evidence of enhancement along the biopsy tract. At surgery, specimens obtained from the biopsy tract, as determined using surgical navigation, revealed GBM. CONCLUSION: Seeding of the biopsy tract, radioresistance and the time interval until radiotherapy are the most likely explanations for tumor growth along the biopsy tract in this case. Consideration should be given for an early start to radiotherapy among those who undergo stereotactic biopsy for GBM. Further research will allow one to determine the radiosensitivity of these tumors and determine which patients may benefit from a radiosurgical or fractionated radiotherapy boost to the biopsy tract. PMID- 11859969 TI - Alterations of INK4a(p16-p14ARF)/INK4b(p15) expression and telomerase activation in meningioma progression. AB - Dysregulation of cell cycle progression and telomerase activation have been implicated in malignant tumor progression as well as in the evasion of senescence and immortalization. We have investigated expression of the cell cycle control and tumor suppressor genes INK4a(p16-p14ARF), INK4b(p15-p10) and RB, and their relation to telomerase activation during malignant meningioma progression. 7/26 (27%) benign, 3/12 (25%) atypical but 4/7 (57%) anaplastic tumors lacked both, p16 and p15 protein expression. 14/39 (36%) benign and atypical but 5/7 (71%) anaplastic meningiomas contained no p14ARF mRNA. 2/46 (4%) tumors failed to express pRB. We observed frequent differential loss of expression of the alternatively spliced INK4a tumor suppressors p16 and p14ARF. Exclusive expression of the alternative INK4b transcript p10 possibly at the expense of p15 and therefore resulting in loss of p15 tumor suppressor activity was noted in two meningiomas. We have previously described telomerase activity or expression of the telomerase catalytic subunit hTERT in this meningioma series. Telomerase activation was detected in 10/27 (37%) benign, but 18/19 (95%) non-benign meningiomas. We observed no significant overall correlation between loss of INK4a/INK4b expression and telomerase activation. In conclusion, our results suggest a greater role for losses of INK4a/INK4b gene products in meningioma formation and malignant progression than previously thought. Inactivation of p16/p15- and pl4ARF-dependent pathways possibly in conjunction with telomerase activation might be critical steps for a meningioma cell towards escape from senescence, that is, immortalization. PMID- 11859972 TI - Horizontal maxillotomy for exposure of the central skull base: the Yale experience. AB - Le Fort I osteotomy allows the surgeon to safely down-fracture the maxilla for wide exposure of the central skull base. This surgical approach is easily extended inferiorly to include the arch of C1, providing 8 cm of horizontal anterior exposure and 5 cm of posterior. Wide operative exposure and a low rate of complications afford superior functional and cosmetic preservation in removing tumors of the central cranial base. PMID- 11859973 TI - Brain metastases from fallopian tube carcinoma responsive to intra-arterial carboplatin and intravenous etoposide: a case report. AB - Fallopian tube carcinoma is the least common neoplasm of the female genital tract. Although rare, neurological complications such as brain metastases can develop. It remains unclear, however, what role chemotherapy has in the treatment of these patients and what route of administration is most effective. Intra arterial (IA) regional administration of chemotherapy may increase intra-tumoral drug concentrations and improve efficacy. We report the case of a 47-year-old woman who developed bilateral fallopian tube cancer and multifocal brain metastases. After progression through radiation therapy and oral chemotherapy, she was placed on IA carboplatin (200 mg/m2/d x 2 days every 4 weeks) and intravenous etoposide (100 mg/m2/d x 2 days every 4 weeks). During treatment she had objective tumor shrinkage that has remained stable for more than 12 months. For patients with fallopian tube carcinoma that develop brain metastases and respond poorly to surgery and/or irradiation, multi-agent chemotherapy containing carboplatin should be considered. The effectiveness of carboplatin may be improved if administered by the IA route. PMID- 11859976 TI - Decreasing incidence and improved survival of laryngeal cancer in Finland. AB - The decreasing incidence rate and improvement in survival of laryngeal cancer patients in Finland are exceptions among western countries. A descriptive study of these trends was conducted including both nationwide population-based cancer registry data with 5 766 patients diagnosed in 1956-1995 and regional hospital based data from Northern Finland, allowing classification into supraglottic and glottic cancers, with 353 patients diagnosed in 1976-1995. In Finland, the age adjusted incidence rate among males decreased from 6.5 per 100 000 in 1956-1965 to 3.5 in 1986-1995, while in females the rate remained around 0.3 per 100 000. The rates in Northern Finland were slightly higher and the supraglottic to glottic incidence ratio diminished from 1.4:1 in 1976-1985 to 0.5:1 in 1986-1995. The 5-year relative survival rate improved in both Northern Finland and the whole country, most noticeably among males and the elderly. In the data from Northern Finland, the survival rate was more favourable in glottic (80%) than in supraglottic cancer (64%). Considering the marked decrease in the incidence of the less favourable supraglottic disease, the observed improvement in survival was small. PMID- 11859975 TI - The prognostic value of tumor markers in patients with glioblastoma multiforme: analysis of 32 patients and review of the literature. AB - Although several studies have examined brain tumor markers for prognostic value, few investigations have stratified analysis based on specific histologic grade. The objective of this study was to evaluate a single histologic grade of glioma, the grade IV glioma or glioblastoma (World Health Organization Classification), with a comprehensive panel of tumor markers in an attempt to identify those with prognostic significance. Tumor samples from a cohort of patients with glioblastoma multiforme (n = 32) were examined for tumor markers, DNA analysis, and clinical variables in an attempt to determine a 'profile' for this tumor. We used univariate and multivariate statistical analysis to determine the prognostic value of tumor cell ploidy, percent S-phase, DNA index, p53, and Ki-67 labeling index, as well as the variables of gender, race, age, location of tumor, history of chemotherapy, and primary versus recurrent tumor. Two additional tumor markers, multidrug resistance gene 1 and glutathione-S-transferase subtype pi, were included in the sample testing, but were not analyzed statistically. Univariate analysis indicated that increasing age had a strong association with decreased survival. Female gender, increasing Ki-67, no chemotherapy before sample collection, and primary glioblastoma showed some association with decreased survival in the univariate model. The univariate results indicated that race, side of tumor, ploidy, S-phase, DNA index, and p53 had no prognostic value. Multivariate modeling demonstrated that age, gender, and Ki-67 were the strongest factors associated with survival. The relevant literature is reviewed. PMID- 11859974 TI - Role of transcription factors in the pathogenesis of pituitary adenomas: a review. AB - The diversity inherent in every organ has its roots in gene-expression variation and is revealed through distinctions in the molecular profile and hence the identity of individual cell type. Study into the molecular mechanisms of the development of individual cell type within the pituitary, which is under the control of transcription factors, has provided a basis for a deeper insight into the molecular mechanisms underlying the pathogenesis of a variety of hormone producing pituitary tumors. Identification of some of these transcription factors in pituitary adenomas further supports their role in the pathogenesis of pituitary adenomas. Understanding the molecular mechanisms of regulation of proliferation of pituitary cell types by transcription factors offers a basis for hope that rational genetic or pharmacologic therapies for pituitary tumors can be designed in the future. PMID- 11859977 TI - Treatment outcome after laser excision of early glottic squamous cell carcinoma- a literature survey. AB - Two treatment options are widely used for the cure of T1 glottic squamous cell carcinoma: radiotherapy and surgical removal. There is ongoing controversy about whether laser excision should be offered to patients with T1 glottic carcinoma. The purpose of this study is to present a review of studies dealing with treatment outcome after laser excision of T1 glottic carcinoma. Eighteen original papers on outcomes were identified. Recurrence rates ranged from 4% to 35%. The disease-specific survival rate at 5 years was found to be from 89% to 100% and crude survival from 74% to 100%. Of the six studies dealing with voice quality, radiation therapy was found to be more effective in preservation of the voice in three, while in the other three studies, no significant difference could be detected. With respect to costs of treatment, in three out of four studies laser surgery was found to be the more economical treatment option. Laser surgery seems to provide comparably low recurrence rates and high disease-specific survival as compared with radiotherapy. In T1 cancer, laser resection leaves the patient with a poorer voice quality than is the case with radiation therapy, but laser treatment seems to be the cheaper option. PMID- 11859978 TI - Squamous cell carcinoma of the nasopharynx--an analysis of treatment results in 149 consecutive patients. AB - The purpose of this study was to evaluate the outcome of primary treatment and treatment of recurrences in patients with nasopharyngeal carcinoma. The material included 149 consecutive patients seen at the Aarhus University Hospital from 1963 to 1991 (49 females and 100 males). The stage distribution was: Stage I-9%, II-3%, III-28%, and IV-60%. Primary treatment was delivered with curative intent in 145 patients (97%). Persistent or recurrent disease after primary radical treatment was observed in 82 of the patients; 54% at the T-level, 40% at the N level, and 33% at the M-level. A curative salvage attempt was carried out in 14 patients only, all with nodal recurrence: surgery in 8 patients (4 controlled) and radiotherapy in 6 patients (2 controlled). The 5-year local tumour control, locoregional tumour control, disease-specific survival rate and the overall survival rate for the patients treated with curative intent were 66%, 53%, 50% and 43%, respectively. Most of the patients (88%) had poorly differentiated tumours and these patients had the best prognosis. A major complication in three patients was radiation-induced myelopathy due to high-dose radiation delivered to the brain stem. Significant positive prognostic factors for treatment outcome in univariate analyses were early T-classification, small clinical stage, poor differentiation and low age. The Cox multivariate analysis showed that early T categories, low N-categories and poor differentiation were independent, positive prognostic factors. Nasopharyngeal carcinoma is curable with primary radiotherapy; patients with poorly differentiated tumours have the best prognosis. Only a few patients were salvaged after recurrence. The factor most essential for success is primary control of the disease at the T- and N-levels. PMID- 11859980 TI - Health-related quality of life of patients with endometrial cancer who are disease-free following external irradiation. AB - Health-related quality of life (HQoL) is assessed through the patients' own evaluation of the impact that a disease and its treatment may have on some of the physical, psychological and social aspects of their lives. The purpose of this study is to describe the HQoL of patients with endometrial cancer who are free of disease after undergoing external irradiation. An HQoL questionnaire was designed and validated, and consisted of the EORTC QLQ-C30 and 80 additional questions. The patients provided self-reported assessments at the end of radiotherapy, and 1, 3, 6, 12, 18 and 24 months later. Forty-nine out of 66 potential subjects participated in the study, which was confined to the period during which the, women were disease free. Most patients experience physical side effects at the end of treatment and up to 6 months thereafter; 10% of the patients have chronic local symptoms and a large number of the patients think about their treatment even two years later. The patients' overall evaluation of their quality of life is lower than that of a matched population of healthy women. PMID- 11859979 TI - Conformal radiotherapy for prostate cancer--longer duration of acute genitourinary toxicity in patients with prior history of invasive urological procedure. AB - The incidence and predictors of acute toxicity were evaluated in patients treated with three-dimensional conformal radiotherapy (3D-CRT) for localized prostate cancer. Between December 1997 and November 1999, 116 patients with T1-T3 prostatic carcinoma were enrolled in the study. Ninety patients were treated with 70 Gy and 26 patients with T3 tumors received 74 Gy. Of the 116 patients 42 (36.2%) had a prior history of invasive urological procedure (IUP) (transurethral resection of the prostate or transvesical prostatectomy for benign prostatic hyperplasia). Acute gastrointestinal (GI) and genitourinary (GU) symptoms were graded according to the EORTC/RTOG scoring system. Toxicity duration after the completion of 3D-CRT was recorded. The majority of patients experienced only mild or no (Grade 1) acute toxicities. Medications for GI and GU symptoms (Grade 2) were required by 28.4% and 12.9% of patients, respectively. Only one case of Grade 3 GI toxicity (0.9%) was observed. Seven patients (6.1%) experienced severe GU toxicity (Grade 3 or 4). No correlation was found between acute toxicity and age, stage, dose (70 Gy vs. 74 Gy), IUP and pelvic lymphadenectomy. A significant relationship was observed between the duration of acute GU toxicity and prior IUP. Symptoms persisted for more than 4 weeks in 51.9% and 26.0% of patients with and without a prior history of IUP, respectively (p = 0.02). The incidence of acute complications, associated with 3D-CRT for prostate cancer, was acceptable in our cohort of patients. A prior history of IUP resulted in a significantly longer duration of acute GU toxicity. PMID- 11859981 TI - Health-related quality of life, anxiety and depression in patients with midgut carcinoid tumours. AB - In earlier studies it has been reported that patients with carcinoid tumours have a relatively good health-related quality of life (HRQoL) and low levels of anxiety and depression. The aims of this study were (a) to investigate the extent to which psychosocial function changes in patients with carcinoid tumours with time from diagnosis and its possible relation to tumour markers, and (b) to compare the HRQoL of patients with carcinoid tumours with that of healthy Swedish adults. Twenty-four patients reported on HRQoL (the EORTC QLQ-C30), anxiety and depression (the Hospital Anxiety and Depression Scale) five times during their first year of treatment. After one year, improvement in nausea/vomiting, flush and anxiety was reported, but there was deterioration of physical function, an increase in muscular pain and problems with dry skin. Levels of tumour markers were not associated with psychosocial function. Patients reported a lower HRQoL compared with healthy Swedish adults. Thus, deterioration of physical function was not accompanied by a deterioration of emotional function, and levels of tumour markers were not related to patients' HRQoL. PMID- 11859982 TI - Body weight loss as an indicator of breast cancer recurrence. AB - Body weight loss (BWL), a major prognostic factor in breast cancer, was included as a parameter to be monitored in the recent breast cancer surveillance guidelines of the American Society of Clinical Oncology. The aim of this work was prospectively to evaluate BWL as an indicator of breast cancer recurrence. Body weight was measured every 2 months for 10.4+/-3.7 (SD) months in 109 disease-free breast cancer patients in stage II node-positive and stage III disease. The correlation between unexplained BWL and recurrence was studied. Attempts were made to define the limits in weight variations among disease-free patients beyond which recurrence could be suspected. Unexplained BWL was observed in 16/19 (84%) patients developing recurrence, versus 9/90 (10%) patients remaining disease free. There was a significant (p < 0.001) correlation between BWL and recurrence. BWL anticipated the diagnosis of recurrence by 6 (range 4-12) months. Based on the average percentage weight variation +/- 2 SD (95% confidence interval) of the disease-free group, the limits for BWL beyond which recurrence could be suspected were a 5.8% decrease in the last 6 months, 3.6% in the last 2 months or 3.0% of the patient's mean weight. However, because of the large variation in the amplitude of individual weight oscillations among disease-free patients (from < 0.5% to > 5.9% of the mean weight), individual limits derived from the patient's own body weight curve seemed more reliable. The results suggest that unexplained BWL is a valuable indicator of incipient breast cancer recurrence. Careful monitoring of body weight in breast cancer patients during follow-up is encouraged. PMID- 11859983 TI - A modified Nottingham prognostic index for breast cancer patients diagnosed in Denmark 1978-1994. AB - Stage of disease at diagnosis is a predictor of breast cancer survival. We used data from the Danish Cancer Register and the Danish Breast Cancer Cooperative Group to study stage distribution in 0-69-year-old Danish breast cancer patients diagnosed in 1978-1994. We constructed a modified Nottingham Prognostic Index (NPI) calculated from the number of excised and positive lymph nodes, malignancy grade and tumour diameter. This NPI could be calculated for 63% of the patients, and among these the stage distribution improved during the study period. The proportion of patients with a poor prognostic score decreased from 27% to 20%. Based on a comparison of the crude 3-year survival of patients with an NPI score and those without, it seems probable that the stage of disease at diagnosis on average improved in Danish breast cancer patients below age 70 during the 1980s and the early 1990s. PMID- 11859984 TI - Breast cancer knowledge, attitudes and practice among nurses in Lagos, Nigeria. AB - A cross-sectional survey was conducted among nurses working in a general hospital in Lagos to determine their knowledge, attitude and practice regarding breast cancer. A self-administered questionnaire was used to investigate knowledge of symptoms, methods of diagnosis and use of cancer screening methods. Two hundred and four nurses out of 280 participated in the study (73% response rate). Knowledge about symptoms, methods of diagnosis and self-breast examination was generally very good. However, only 30% had had a clinical breast examination and 8% a mammogram within the past three years. Use of cancer screening methods was significantly associated with knowledge of the subject (p = 0.03). Twenty-eight percent did not know how to estimate the risk of cancer and 61% believed they were not at risk. Nurses possess adequate knowledge about breast cancer but they need more information on cancer risk estimation. PMID- 11859985 TI - Long-term impact of reproductive factors on the risk of cervical, endometrial, ovarian and breast cancer. AB - The influence of maternal age, parity, low or high birthweight, multiple births, and pre-eclampsia on the risk of cervical, endometrial, ovarian and breast cancers was studied. Data on 40951 women and the outcomes of their deliveries between 1955 and 1995 were obtained from birth registers. For the mothers, data from the Swedish Cancer Registry and the Cause of Death Register were added. The sample was evaluated using Cox's regression in univariate and bivariate analyses where the relative risk and its 95% confidence interval were calculated. Increasing maternal age at first birth was associated with an increasing relative risk of endometrial, ovarian, and breast cancers, and with a decreased risk of cervical cancer. Multiparity was a protective factor for all gynaecological cancers, including cervical and breast cancers. Multiple births were associated with an increased risk of endometrial cancer. PMID- 11859986 TI - Non-infusional 5-fluorouracil, doxorubicin and cisplatin in the treatment of locally advanced or metastatic gastro-oesophageal adenocarcinoma. AB - To reduce the Hickman line-associated morbidity of continuous infusion 5 fluorouracil combined with epirubicin and cisplatin (ECF) and to investigate the need for infusional regimens, we conducted a retrospective study in patients with advanced gastro-oesophageal adenocarcinoma. Thirty-six patients, with histologically proven irresectable gastro-oesophageal adenocarcinoma were given: 60 mg/m2 cisplatin on day 1, 35 mg/m2 doxorubicin on day 1 and 500 mg/m2 5 fluorouracil on days 1 and 8 (NIACF) every 3-weeks. A median of 3 cycles was administered. The principal toxicity was myelosuppression with grade III/IV neutropenia in 47% of cycles. Neutropenic fever occurred in 5% of the cycles: non haematological toxicity was mild and there were no treatment-related deaths. Administered dose intensity was 96.1% for doxorubicin, 93.6% for cisplatin and 90.5% for 5-fluorouracil. There were 16 partial responses and 1 complete response (overall response rate 47%, 95% confidence interval CI 31-63%); 8 patients had stable disease. Median progression-free and overall survival rates were 5 months (95% CI 4-6) and 8 months (95% CI 6-10), respectively. NIACF is a well-tolerated regimen in advanced gastro-oesophageal adenocarcinoma that precludes the need for central venous access, with activity similar to that observed with ECF. PMID- 11859987 TI - Non-constant tumour blood flow--implications for therapy. AB - In the past few years, 'perfusion-limited' hypoxia caused by intermittent decreases in tumour blood flow has received increasing attention. Little effort, however, has gone into characterizing the nature, magnitude or duration of these changes, or their functional significance other than as modifiers of radiotherapy. We have therefore undertaken multiple, quantitative analyses of tumour blood flow in human tumour xenograft systems, and rigorously examined the ramifications of transient blood flow changes. Tumour blood flow in these experimental tumours is much less constant than has previously been assumed, and not only impacts on response to radiotherapy and chemotherapy, but also on the more fundamental processes of tumour growth and repopulation. Notably, responses entirely consistent with the laboratory results have been seen in our initial studies of human tumours sequentially biopsied during treatment. PMID- 11859988 TI - Radiosensitizing effect of carbogen breathing during pulsed irradiation of the rat R1H tumor. AB - The objective of the present study was to investigate the radiosensitizing effect of carbogen breathing during pulsed x-ray irradiation in an experimental tumor model. Rat R1H rhabdomyosarcoma tumors were irradiated with 36 Gy total dose in 1 Gy high dose rate pulses, either hourly repeated, or in an 'office hours' protocol with irradiation-free overnight intervals. With the hourly, pulsed irradiation scheme, tumor growth delay (TGD) was significantly increased from 24.4+/-0.7 days in air-breathing animals to 29.0+/-0.9 days in animals breathing carbogen during irradiation. With irradiation during office hours, the TGD was shortened, and carbogen was less effective. The data show that carbogen acts as a radiosensitizer when applied during pulsed irradiation. Translation of the experimental data to clinical practice indicates that hyperoxygenation of the tumor during pulsed dose rate (PDR) or high dose rate (HDR) brachytherapy might enhance the tumor response of patients. PMID- 11859989 TI - Comments to the article 'Immunohistochemical study of ceruloplasmin, lactoferrin and secretory component expression in neoplastic and nonneoplastic thyroid gland diseases' by A. Kondi-Pafiti et al. in Acta Oncologica 2000; 39 (6) 753-6. PMID- 11859990 TI - Measurement of serum CA 19-9 in biliary diseases requires great caution. PMID- 11859991 TI - Serum elevation of CEA and CA 19-9 in an aggressive variant of prostatic carcinoma. PMID- 11859992 TI - Oxaliplatin-induced haematological emergency with an immediate severe thrombocytopenia and haemolysis. PMID- 11859993 TI - Avoiding the "any models trap". PMID- 11859994 TI - Fathers with serious mental illnesses: a neglected group. AB - In recent years, policy makers, researchers, and practitioners alike have come to recognize how the traditional emphasis on mothers has overshadowed the equally critical contribution of fathers to child development and family life. In spite of this growing recognition of the importance of fathers, the need to better understand and support fathers with serious mental illnesses (SMI) has received little attention within mental health, public welfare, child protection, or criminal justice systems. In an effort to counter the continuing neglect of fathers with SMI, this paper provides an overview of the scant existing research focusing on fathers with SMI and sets forth a series of recommendations highlighting the need for greater inclusion of fathers in future research and ways to support men with SMI in their role as parents. PMID- 11859996 TI - Landlords as partners for promoting success in supported housing: "it takes more than a lease and a key". AB - This paper examines the potential value of working with landlords and property managers to promote success in supported housing for people with psychiatric and addictive disabilities. The authors argue that relationships with landlords can promote tenants' housing stability, rehabilitation, social integration, and success in community living. A case history of the program is used to describe efforts to develop working relationships with landlords. It concludes presenting a model for how tenants, housing programs, and service providers can collaborate with landlords. PMID- 11859995 TI - Measuring practitioners' beliefs, goals, and practices in psychiatric rehabilitation. AB - This article reports on a scale to measure the psychiatric rehabilitation beliefs, goals, and practices of staff who provide services to consumers. The scale's reliability, validity, and factor structure are presented based upon 469 staff members and 191 people in rehabilitation. The scale appears to be a stable measure of staff members' knowledge of modern psychiatric rehabilitation beliefs, goals, and practices as elaborated by the field's leadership. It also appears to provide a valid measure of staff members' actual practice patterns as they relate to the consumer outcomes of empowerment, quality of life, independent living, and competitive employment. Consumers, program administrators, educators, researchers, and practitioners may find the scale useful as a measure of some of the beliefs, goals, and practices that currently define modern psychiatric rehabilitation. PMID- 11859997 TI - The recovery vision for mental health services and research: a British perspective. AB - It may seem logical that if people can break down or become ill, then they can also overcome their problems and recover. Yet providers of mental health services often fail to emphasise such positive possibilities for people with mental health problems-particularly those with the most severe diagnoses. Psychiatric services often emphasize maintenance rather than recovery, and many survivors report that receiving a psychiatric label has been severely detrimental to their efforts to lead a worthwhile and enjoyable life and contribute to others. PMID- 11859999 TI - Painting with a broad brush: professional careers for artists with psychiatric disabilities. AB - Being a professional artist is a possible career for people with psychiatric disabilities; however, this vocation traditionally has been associated with mystique that has not always been complimentary. One program that is working to mitiigate the negative impact of this mystique while promoting professional and as a vocational option is the Anchor House of Artists. The Anchor House, located in Northampton, MA, supports and guides professional artists with Psychiatric disabilities from Massachusetts and the surrounding area. This article includes personal accounts of consumers who have benefited from Anchor House. PMID- 11859998 TI - Aging with a serious mental disability in rural Northern Ontario: family members' experiences. AB - As people with serious mental disabilities grow older, their primary caregivers continue to be family members. This qualitative study explored the life experiences of people aging with a preexisting serious mental disability living in rural Northern Ontario, from the perspective of family members, and it identified the factors which facilitated or impeded their relative's current and future participation in community life. Results included the challenges of rural life, lack of support by health care providers, rural service needs, caregiver responsibility, the impact of aging on the individual and the family, and future concerns. New models for effective rural programs, services, community education, and effective partnerships with family members must be investigated. PMID- 11860000 TI - Culture, control, and family involvement: a comparison of psychosocial rehabilitation in India and the United States. AB - Research has shown better prognosis rates for people with psychiatric disabilities in developing countries than in Western countries. By comparing psychosocial rehabilitation in India and the United States, this paper explores how cultual beliefs and practices impact the recovery of people with psychiatric disabilities. In India, an emphasis on interdependence, externalized locus of control, and family involvement, are significantfactors in caring for people with psychiatric disabilities. In contrast, rehabilitation goals in the United States focus upon independence and individual productivity. The paper examines the implications of these findings and the extent to which they can be applied across cultures. PMID- 11860001 TI - Identifying consumer-provider benefits in evaluations of consumer-delivered services. AB - Consumer-delivered services are different in many ways from traditional mental health services and require unique approaches to how they are studied. This includes attending to benefits to both consumer-providers as well as to program participants. A qualitative study was conducted to systematically examine consumer-provider benefits. A thematic analysis of interviews with 14 peer providers from Friends Connections, a peer-support program for persons with recurring mental health and substance use disorders, was conducted. Responses indicate that peer providers benefit from their roles as helpers, a finding consistent with the helper-therapy principle. Implications for research and policy are discussed. PMID- 11860002 TI - Differences in job retention in a supported employment program Chinook Clubhouse. AB - This is a retrospective study comparing two groups of adults with severe mental illnesses in a clubhouse program, who were employed from October 1995 to June 1999. Long-Term Workers, (n = 22) were those who have held a job for over one year duration, while Short-Term Workers, (n = 20) were those who were not able to hold a job for more than a year. Data was gathered through focus groups, and individual interviews. Statistical data found the LTW group overall less disabled, possessing a positive attitude and setting more realistic vocational goals. PMID- 11860003 TI - Outcome measures across program sites for postsecondary supported education programs. AB - This article briefly describes three supported education programs and examines outcomes for each. The program settings were a mental health center, a clubhouse, and a community college. Students (n = 124) were followed for five semesters to assess program outcomes. Although this study did not statistically control for variations in services among sites, each site adhered to the principles and practice of supported education as a specialized intervention. Differences among sites in student demographics, education and employment outcomes, satisfaction with school, job/education fit, satisfaction with life, and self-esteem are reported. Although there were variations in outcomes among sites, few signifcant differences were found. PMID- 11860004 TI - Mental health experiences and deafness: personal and legal perspectives. AB - The following study describes the experiences of Ms. Jan DeVinney, a woman with deafness, who voluntarily admitted herself to the psychiatric unit of a general hospital seeking help for depression. Presented are her self-reported, detailed experiences that reveal many of the accessibility barriers that face deaf people within mental health settings. Also included are the provisions of a nationally significant consent decree emanating from DeVinney and the United States of America vs. Maine Medical Center. Within the landmark consent decree are contained specific hospital reforms illustrating what is required to achieve inclusive mental health services for deaf and partially hearing people. The study also reveals broader issues surrounding mental health treatment that are relevant to services recipients, whether deaf or hearing. PMID- 11860005 TI - The goals of psychiatric rehabilitation: an ethical analysis. AB - This study addressed the question under what conditions ought client-centered psychiatric rehabilitation work towards goals set by its clients. This was done by means of an ethical analysis of goal setting in psychiatric rehabilitation, using standard ethical principles of health care and a case study for illustration. The analysis demonstrated that client-centered psychiatric rehabilitation ought to work towards goals set by its clients under the conditions that they not involve serious danger and are not induced by severe mental impairment. When these conditions are not fulfilled, it may be most helpful to engage the clients in critical dialogue so as to reach an agreement regarding the setting of goals for their psychiatric rehabilitation. Special ways of engaging clients with severe mental impairments in such dialogue may have to be explored. PMID- 11860006 TI - The story of my recovery. AB - Chuan-Lin Alice Tsai is the Consumer Representative at Stepping Stones of Rockford, Inc., a community mental health agency in northern Illinois that provides rehabilitation and housing for adults with mental illness. "Consumers" are people with a diagnosis of mental illness or people who are receiving services for mental illness. She served as Chair of the Consumer Family Forum Spring Conference in 2001 and Vice-Chair of the Consumer/Family Forum in 2000 and 2001. The Consumer/Family Forum of the Northwest Network is the consumer, family, and provider organization for the nine northern counties of Illinois. PMID- 11860007 TI - I have a life plan. PMID- 11860008 TI - An auxotrophic pigmented Cryptococcus neoformans strain causing infection of the bone marrow. AB - Cryptococcosis, caused by an encapsulated fungus, Cryptococcus neoformans, has emerged as a life-threatening infection in HIV-positive individuals and other immunocompromised hosts. This report describes an unusual strain of C. neoformans isolated from an AIDS patient that developed pigment on Sabouraud's medium. The yeast was auxotrophic for adenine due to a deletion in the coding region of ADE2, and was complemented by introduction of a functional copy of the ADE2 gene from C. neoformans. The yeast had an unusual myelotropism that was clinically evident as a pancytopenia with displacement of bone marrow precursors by yeast cells, and it had an unusual spectrum of infection in the human host. This is the first description of a nutritional auxotroph of C. neoformans isolated from a patient. PMID- 11860009 TI - Chemiluminescence of superoxide generated by Candida albicans: differential effects of the superoxide generator paraquat on a wild-type strain and a respiratory mutant. AB - We previously reported that a respiration-competent parent strain (K) of Candida albicans was more susceptible to the intracellular superoxide radical (O2-) generator paraquat (PQ) than was a respiration-deficient mutant (KRD-19), although both showed a similar sensitivity to extracellularly generated O2-. To clarify the cause of the differential PQ lethality, we developed a chemiluminescence method for measuring O2- generated by C. albicans cells by using the probe methyl-Cypridina-luciferin analogue (MCLA), and examined the effects of PQ on O2- generation in both parent and mutant strains. Endogenous O2- generation without stimulation by PQ was unexpectedly low in both strains. PQ induced O2- generation in the parent strain was maximal in logarithmic phase cells and lowered in stationary phase cells. In contrast, O2- generation in the mutant remained low throughout the growth phase, even when stimulated by PQ. The extent of PQ-induced O2- generation in the parent strain depended on the carbon source added to the assay mixture; in decreasing order, glucose, glycerol, no carbon source. The inhibitor of the cytochrome respiratory chain, antimycin A, suppressed almost completely the PQ-induced O2- generation in the parent strain. It has been established that PQ is converted to its radical form (PQ+) by receiving a single electron in cells. PQ+ then reduces molecular oxygen to O2- by redox cycling. Thus, the high tolerance to PQ of the respiration-deficient mutant can be explained by minimal PQ+/O2- production due to the limited supply of electrons from the impaired respiratory system. PMID- 11860010 TI - Effect of granulocyte colony-stimulating factor and granulocyte-macrophage colony stimulating factor on polymorphonuclear neutrophils, monocytes or monocyte derived macrophages combined with voriconazole against Cryptococcus neoformans. AB - The antifungal activity of voriconazole (VCZ) was tested against Cryptococcus neoformans (Cn) with and without the addition of polymorphonuclear neutrophils (PMN), monocytes or monocyte-derived macrophages (MDM) in vitro. Human effector cells with and without the addition of VCZ were incubated with Cn for 24 h. PMN, mono and MDM alone resulted in 61%, 34% and 23% inhibition of Cn, respectively (n = 3, P<0.01). VCZ at 0.01 and 0.05 microg ml(-1) alone resulted in 48% inhibition and 19% killing (n = 6). The addition of VCZ at 0.01 and 0.05 microg ml(-1) to human effector cells enhanced killing of Cn by 51% and 71% for the PMN, 41% and 58% for the mono, and 14% and 34% for the MDM, respectively. The addition of either granulocyte colony-stimulating factor (G-CSF) or granulocyte-macrophage colony stimulating factor (GM-CSF) significantly enhanced the ability of human effector cells to kill Cn. G-CSF and GM-CSF plus PMN resulted in 47% and 46% killing, respectively; GM-CSF plus monocytes or MDM resulted in 31% or 22% killing, respectively. G-CSF and GM-CSF further enhanced the collaborative killing effect of human effector cells and VCZ. At 0.01 and 0.05 microg ml(-1) of VCZ, G-CSF or GM-CSF enhanced PMN killing to 92% and 93% or 87% and 94%, respectively. GM-CSF enhanced both mono and MDM with VCZ at 0.01 and 0.05 microg ml(-1) in killing Cn to 62% and 86%, and 61% and 84%, respectively. These results suggest that VCZ would have good efficacy in the treatment of Cn infection in humans. Furthermore, VCZ would have enhanced efficacy in clinical settings where either G-CSF or GM-CSF was being used. PMID- 11860011 TI - Immobilization of Aspergillus fumigatus colonies in a soft agar matrix allows visualization of A549 cell detachment and death. AB - A model to examine the effects of proliferating Aspergillus fumigatus ATCC 26933 colonies embedded in a thin layer of soft agar on a monolayer of A549 cells was developed and enabled an investigation of the response of cultured cells to fungal growth. This model simulates the conditions on the respiratory surface in patients with invasive aspergillosis and also in the mucus secretions of cystic fibrosis patients with allergic bronchopulmonary aspergillosis. Conidia of A. fumigatus adhering to A549 cells were immobilized in a thin layer of soft agar (0.6% (w/v)) and allowed to germinate at 37 degrees C. Fungal colonies greater than 5 mm in diameter caused rounding-up and detachment of A549 cells underneath the colony and towards the hyphal tips. As the fungal colony diameter increased, cell detachment occurred ahead of the hyphal tips. Cells detached for short periods (less than 6 h) showed no annexin-V (AV) or propidium iodide (PI) staining, suggesting no externalization of phosphatidylserine and an intact plasma membrane. Cells that had detached for periods greater than 6 h were positive for AV and PI indicating the rupture of the plasma membrane and cell death by necrosis. Chemical extraction and separation by thin layer chromatography of agar from zones of cell detachment around fungal colonies revealed the presence of three compounds that may play a role in inducing cell death. PMID- 11860012 TI - Purification and characterization of urease isolated from the pathogenic fungus Coccidioides immitis. AB - Coccidioides immitis, the causative agent of San Joaquin Valley fever (coccidioidomycosis), produces a urease which has been suggested to contribute to the virulence of this fungal pathogen. Urease catalyzes the hydrolysis of urea and has been proposed to at least partly account for alkalinity of the microenvironment in which C. immitis grows due to the release of ammonia and ammonium ions. The C. immitis urease was purified to homogeneity (1048-fold) from the mycelial cytosol by chromatographic fractionation. The sequence of 12 N terminal amino-acid residues of the purified, native polypeptide was identical to that predicted by the translated urease gene sequence which has been reported. The isolated enzyme exhibited a specific activity in the presence of urea of 1750 micromol min(-1) mg(-1) protein, has a native molecular mass of 450 kDa, revealed a Km for urea of 4.1 mM, had a pH optimum of 8.0 and is heat stable. Hydroxyurea, acetohydroxamic acid (AHA) and boric acid each inhibited activity of the purified enzyme. Urease activity was enhanced by the presence of 5-10 mM concentrations of Mg2+ or Mn2+, but inhibited by Li+, Ni2+, Cu2+ or Zn2+. The reversible urease inhibitor, AHA, blocked enzyme activity in the crude mycelial cytosolic fraction when added at a concentration of 10 mM. On the other hand, 10 mM AHA added to 4 day-old mycelial cultures only partially decreased the amount of ammonium detected in the culture medium. It is evident, therefore, that C. immitis urease activity does not account for the total amount of ammonia secreted during in vitro growth of the pathogen. Other metabolic sources of ammonia, which may also contribute to the virulence of C. immitis, are under investigation. PMID- 11860013 TI - Environmental sampling for Cryptococcus neoformans var. gattii from the Blue Mountains National Park, Sydney, Australia. AB - The environmental association of Cryptococcus neoformans var. gattii with decaying wood in tropical and subtropical regions of the world is well documented. In Australia, the yeast appears confined to certain species of Eucalyptus or very closely related tree species. In this study, we attempted to isolate C. n. var. gattii from different gum tree species in the Blue Mountains National Park. Out of 99 samples from 9 different tree species, only 3 yielded viable yeast colonies; 2 were from turpentine gums (Syncarpia glomulifera) and 1 was from a decayed stump of an unknown species. All of the colonized trees occurred in close proximity in urbanized areas of the Park, and all isolates shared identical DNA fingerprinting profiles. We suggest that domestic animal vectors may be responsible for the introduction and transmission of the yeast in this region, but that propagation and dispersal are very limited. This study indicates that C. n. var. gattii may occur on trees and in areas that were not previously expected to host it. However, the low incidence in the Blue Mountains National Park means this yeast is unlikely to pose any hazards to humans and animals living in or visiting this area. PMID- 11860014 TI - Molecular cloning of an extracellular aspartic proteinase from Rhizopus microsporus and evidence for its expression during infection. AB - An extracellular aspartic proteinase (Rmap) from Rhizopus microsporus var. rhizopodiformis was detected in the culture supernatant of a fungal isolate from a case of rhinocerebral mucormycosis (case HA). The proteinase was purified to near homogeneity by ion exchange and affinity chromatography on pepstatin agarose. Based on its N-terminus the RMAP gene was cloned and found to code for 388 amino acids. The preproenzyme has an aminoterminal leader sequence of 65 amino acids, whereas the mature enzyme consists of 323 amino acids. The deduced amino-acid sequence of the preproenzyme was 82% homologous to an extracellular aspartic proteinase of Rhizopus niveus. Low stringency Southern blot analysis of R. microsporus DNA suggested the presence of other homologous genes. Expression of Rmap in Pichia pastoris was achieved, and the recombinant enzyme was active in the yeast culture supernatant. Both enzyme preparations exhibited a similar optimum of activity in the pH 2.5 region. Furthermore, Rmap was shown to activate bovine blood coagulation factor X at slightly acidic pH in vitro. Expression of the proteinase during mycosis was proven by a specific immune response of patient HA. PMID- 11860015 TI - Cryptococcal meningitis caused by Cryptococcus neoformans var. gattii, serotype C, in AIDS patients in Soweto, South Africa. AB - We present four patients from South Africa with meningitis caused by Cryptococcus neoformans var. gattii, serotype C. These are the first patients with human immunodeficiency virus (HIV) infection to be reported with serotype C meningitis. PMID- 11860016 TI - Transcription initiation of genes associated with azole resistance in Candida albicans. AB - Oral infections with the opportunistic pathogenic yeast Candida albicans are one of the earliest and most frequent infections in immunosuppressed individuals. In these populations, drug-resistant isolates have emerged with the widespread use of antifungal azole drugs. Many molecular mechanisms of resistance have been identified, including overexpression of two types of efflux pumps, the major facilitator MDR1 and the ABC-transporters (CDR1 and CDR2), and the overexpression or mutation of the target enzyme, ERG11. With overexpression of these four genes implicated in multidrug resistance, identification of regulatory regions of the promoters is important. 5' rapid amplification of cDNA ends (RACE) was used to identify transcription initiation sites for genes associated with multidrug resistance (CDR1, CDR2, MDR1 and ERG11). These results were confirmed by cloning and sequencing of 5' RACE products and by primer extension. This research will allow further analysis of the regulation of transcription for these genes. PMID- 11860017 TI - Utilization of the internal transcribed spacer regions as molecular targets to detect and identify human fungal pathogens. AB - Advances in molecular technology show great potential for the rapid detection and identification of fungi for medical, scientific and commercial purposes. Numerous targets within the fungal genome have been evaluated, with much of the current work using sequence areas within the ribosomal DNA (rDNA) gene complex. This section of the genome includes the 18S, 5.8S and 28S genes which code for ribosomal RNA (rRNA) and which have a relatively conserved nucleotide sequence among fungi. It also includes the variable DNA sequence areas of the intervening internal transcribed spacer (ITS) regions called ITS1 and ITS2. Although not translated into proteins, the ITS coding regions have a critical role in the development of functional rRNA, with sequence variations among species showing promise as signature regions for molecular assays. This review of the current literature was conducted to evaluate clinical approaches for using the fungal ITS regions as molecular targets. Multiple applications using the fungal ITS sequences are summarized here including those for culture identification, phylogenetic research, direct detection from clinical specimens or the environment, and molecular typing for epidemiological investigations. The breadth of applications shows that ITS regions have great potential as targets in molecular-based assays for the characterization and identification of fungi. Development of rapid and accurate amplification-based ITS assays to diagnose invasive fungal infections could potentially impact care and improve outcome for affected patients. PMID- 11860018 TI - Unusual traits of the pyoverdin-mediated iron acquisition system in Pseudomonas putida strain BTP1. AB - Fluorescent Pseudomonas species are characterized by the production of pyoverdin type siderophores for Fe3+ acquisition in iron-limited environments. Since it produces a structurally specific pyoverdin, Pseudomonas putida strain BTP1 could represent a valuable tool in an attempt to correlate the structural features of these compounds with some specificity in their two main properties i.e. affinity for iron and recognition rate by other Pseudomonas strains. An uncommonly high affinity for iron of the pyoverdin synthetized by P. putida BTP1 was observed by comparing both the apparent stability constant and the decomplexation kinetic of its ferric complex with those of ferripyoverdins from other strains. On another hand, results from growth stimulation experiments and labeled ferripyoverdin uptake assays highlighted the very low recognition rate of BTP1 isopyoverdins by membrane receptors of foreign strains. By contrast, P. putida BTP1 was able to utilize a broad spectrum of structurally unrelated exogenous pyoverdins by means of multiple receptors that are likely constitutively expressed in its outer membrane. The unusual traits of its pyoverdin-mediated iron acquisition system should contribute to enlarge the ecological competence of Pseudomonas putida BTP1 in terms of colonization and persistence in the rhizosphere. PMID- 11860019 TI - Cadmium-induced apoptosis in C6 glioma cells: mediation by caspase 9-activation. AB - The induction of apoptotic cell death by cadmium was investigated in eight mammalian cell lines. Great differences in the cytotoxicity of cadmium were found with different cell lines: Rat C6 glioma cells turned out to be most sensitive with an IC50-value of 0.7 microM, while human A549 adenocarcinoma cells were relatively resistant with an IC50-value of 164 microM CdCl2. The mode of cadmium induced cellular death was identified to involve apoptotic DNA fragmentation in three cell lines, i.e., in C6 glioma cells, E367 neuroblastoma cells and NIH3T3 fibroblasts. In C6 glioma cells, this process was investigated in detail. Internucleosomal DNA-fragmentation occurred 40 h after application of CdCl2 and was concentration-dependent between 1-100 microM CdCl2, followed by a decrease at higher concentrations due to necrotic processes. Apoptotic chromatin-condensation and nuclear fragmentation was observed 48 h after application of 2.5 microM CdCl2. Furthermore, cadmium (1 microM, 48 h) caused a breakdown of the mitochondrial membrane potential as shown by the decline in mitochondrial uptake of rhodamine 123. Also, we found an activation of caspase 9, a protease known to be activated in apoptotic processes following mitochondrial damage. Besides Cd2+, other toxic heavy metal ions (Hg2+, Pb2+, Ni2+, Fe2+, CrO4(2-), Cu2+ or Co2+) did not induce apoptotic DNA fragmentation in C6 cells. The only exception was Zn2+ which caused apotosis at high concentrations (>150 microM) whereas it protected against cadmium-induced apoptosis at low concentrations (10-50 microM). PMID- 11860020 TI - pH dependent conformational changes within the iron responsive element. AB - The expression of several mRNAs related to iron importation, storage and utilization within mammalian cells are regulated through interactions of iron regulatory proteins with an iron responsive element, an RNA hairpin with a bulged C. A dimethylsulfate modification interference assay was used to demonstrate that the iron responsive element undergoes significant pH dependent conformational changes. Specifically, it was demonstrated that the phylogenetically conserved A within the hairpin loop and an intra-loop C-G base pair are highly sensitive to changes in pH. The conserved C of the bulged loop does not significantly affect the pH dependent conformational changes of the hairpin loop. These studies have structural implications for an RNA-protein interaction that is critical to mammalian iron regulation. PMID- 11860021 TI - Abnormal iron delivery to the bone marrow in neonatal hypotransferrinemic mice. AB - Hypotransferrinemic (HP) mice have a splicing defect in the transferrin gene, resulting in < 1% of the normal plasma levels of transferrin. They have severe anemia, suggesting that transferrin is essential for iron uptake by erythroid cells in the bone barrow. To clarify the significance of transferrin on iron delivery to the bone marrow, iron concentration and 59Fe distribution were determined in 7-day-old HP mice. Iron concentration in the femur, bone containing the bone marrow, of HP mice was approximately twice higher than in wild type mice. Twenty-four h after injection of 59FeCl3, 59Fe concentration in the bone and bone marrow of HP mice was also twice higher than in wild type mice. The present findings indicate that iron is abnormally delivered to the bone marrow of HP mice. However, the iron seems to be unavailable for the production of hemoglobin. These results suggest that transferrin-dependent iron uptake by erythroid cells in the bone marrow is essential for the development of erythrocytes. PMID- 11860022 TI - In vitro binding of inorganic mercury to the plasma membrane of rat platelet affects Na+-K+-Atpase activity and platelet aggregation. AB - Hg2+ binding to ouabain-sensitive Na+-K+-ATPase of rat platelet membrane was specific with a Ka of 1.3 x 10(9) moles and Bmax of 3.8 nmoles/mg protein. The binding of mercury to Na+-K+-ATPase also inhibits the enzyme significantly (P < 0.001), which is greater than its ouabain sensitivity. Further in the cytosol of washed platelets conjugation of reduced glutathione (GSH) to Hg2+ is correlated dose dependently (25, 50 and 100 pmoles) to enhanced GSH-S-transferase (GST) activity. It may be concluded from the present in vitro experiments that mercury binds specifically to thiol groups present in the platelet membrane Na+-K+ ATPase, inhibits the enzyme and induces changes in platelet function, namely, platelet aggregation by interfering with the sodium pump. PMID- 11860023 TI - Two pathways of iron uptake in bovine spleen apoferritin dependent on iron concentration. AB - Iron incorporation by bovine spleen apoferritin either with ferrous ammonium sulfate in different buffers or with ferrous ammonium sulfate and phosphate was studied. Iron uptake and iron autoxidation were recorded spectrophotomerically. The buffers [4-(2-hydroxyethyl)-1-piperazinyl]ethanesulphonic acid (Hepes) and tris(hydroxymethyl)aminoethane (Tris) exhibited pH-dependent iron autoxidation, with Tris showing less iron autoxidation than Hepes. An Eadie-Scatchard plot (v/[s] versus v) of the iron uptake rate in Hepes was a curved rather than a straight line, suggesting that there are two iron uptake pathways. On the other hand, the Eadie-Scatchard plots of Tris and of Hepes after the addition of phosphate showed a straight line. Phosphate accelerated the iron uptake rate. The iron loading kinetics of apoferritin in Hepes was dependent on apoferritin concentration. The Km value obtained from iron uptake kinetics was 4.5 microM, corresponding to the physiological iron concentration. These results demonstrate that iron loading of apoferritin was accomplished at physiological iron concentrations, which is essential for iron uptake, via two uptake pathways of dependent on iron concentration. PMID- 11860024 TI - Effects of molybdenum on fertility of male rats. AB - Sodium molybdate was administered orally to adult male rat at dose level of 10, 30, and 50 mg kg body weight (5 days per week) for 60 days. At higher dose levels significant decrease in absolute and organ-to-body weight ratios of testes, epididymides, seminal vesicles and ventral prostate was observed. The sperm abnormality, associated with decrease in sperm motility and sperm count was also observed. Significant alterations in the activities of marker testicular enzymes, viz. sorbitol dehydrogenase (decreases), lactate dehydrogenase (increases) and gamma-glutamyl transpeptidase (increases) associated with histopathological changes in testes was also observed. Accumulation of molybdenum in testes, epididymides and seminal vesicles was also observed. The study reveals that the oral ingestion of molybdenum may affect the histoarchitecture of testes and sperm morphology. The testicular and spermatotoxic changes may be responsible for observed male mediated developmental toxic effects. PMID- 11860025 TI - Ferricrocin--an ectomycorrhizal siderophore of Cenococcum geophilum. AB - The ectomycorrhizal fungus Cenococcum geophilum was grown in low-iron medium and the excreted siderophores were extracted, purified and analyzed by HPLC. The principal hydroxamate siderophore produced, was identified as ferricrocin as confirmed by analytical HPLC, FAB-mass spectrometry and 1H- and 13C-NMR spectra. Although the occurrence of ferricrocin has been shown earlier to occur in the ericoid mycorrhizal fungi, this is the first report of ferricrocin in a true ectomycorrhizal fungus which is taxonomically related to the ascomycetes. PMID- 11860026 TI - Copper(II) complexes of the beta-blocker pindolol: properties, structure, biological activity. AB - The complex formation between copper(II) and the antihypertensive drug pindolol (HPin) was studied both in aqueous and methanolic media. Two complexes are formed at different metal-to-ligand molar ratios. The mononuclear complex Cu(Pin)2(HPin)2 contains two ligands in an anionic bidentate form and two--in a neutral form bound monodentately. The second complex Cu2Pin2Cl2 is dinuclear and its structure was determined by X-ray diffraction. The compound crystallizes in the monoclinic group C2/c with cell components a = 14.4998(13)A, b = 18.511(2)A, c = 14.2982(13)A, alpha = 90 degrees, beta = 109.556(2) degrees, gamma = 90 degrees and Z = 12 at 293K. A pharmacological study on the influence of pindolol and its mononuclear complex on the heart rate of rats was performed. The complex is more active and has a longer effect in comparison with the pure non coordinated pindolol in equitoxic doses. PMID- 11860027 TI - Correlations of trace element levels within and between different normal autopsy tissues analyzed by inductively coupled plasma atomic emission spectrometry (ICP AES). AB - Imbalance in trace metal metabolism may lead to metal interactions that may be of patho-physiological importance. Knowledge of the relation between trace metals in normal tissues is needed to assess abnormal deviations associated with disease. In this study correlations between Cu, Co, Cr, Fe, Mn, Ni, Se, Zn, Al, Ba, Cd, Pb and Sr within the same and between 6 different, normal autopsy tissues were determined using Spearman rank correlation analysis based on analytical data obtained by inductively coupled plasma atomic emission spectrometry (ICP-AES). Fe Co were correlated in most tissues. Cu-Mn, Zn-Cu, Zn-Mn and Zn-Cd were highly correlated in the kidney medulla. Ni-Ni, Sr-Sr and Cd-Cd were correlated between several tissues, while Fe-Fe, Zn-Zn and Cu-Cu were correlated between kidney cortex and medulla. Mn-Mn was highly correlated between the liver and brain front lobe, cerebellum and heart. High correlations were found for Ni-Co and for Se-Mn between the kidney cortex and brain front lobe and pancreas respectively. Inverse correlations were found for Se-Cd between kidney cortex and cerebellum, for Se-Cd and Cd-Zn between kidney medulla and heart, for Co-Sr and Fe-Sr between the liver and kidney cortex and heart respectively, and for Sr-Mn between kidney medulla and pancreas. A large number of trace elements are statistically correlated within and between different, normal tissues. Knowledge of these correlations may contribute to increase the understanding of kinetic interactions of trace metals in the body and the role of such interactions in normal and disturbed trace metal metabolism. PMID- 11860028 TI - Characterization of polypeptide antibiotics of the polymyxin series by liquid chromatography electrospray ionization ion trap tandem mass spectrometry. AB - A selective reversed phase liquid chromatography/mass spectrometry (LC/MSn) method is described for the identification of related compounds in commercial polymyxin B samples. Mass spectral data for these polypeptide antibiotics were acquired on a LCQ ion trap mass spectrometer equipped with an electrospray ionization probe operated in the positive ion mode. The LCQ ion trap is ideally suited for the identification of the related substances because it provides on line LC/MSn capability. The main advantage of this hyphenated LC/MSn technique is the characterization of novel related substances without time-consuming isolation and purifications procedures. Using this method six novel related substances were partially identified in a polymyxin B bulk sample. PMID- 11860029 TI - Probing structural requirements of fMLP receptor: on the size of the hydrophobic pocket corresponding to residue 2 of the tripeptide. AB - The conformationally constrained f-L-Met-Ac(n)c-L-Phe-OMe (n = 4,9-12) tripeptides, analogues of the chemoattractant f-L-Met-L-Leu-L-Phe-OH, were synthesized in solution by classical methods and fully characterized. These compounds and the published f-L-Met-Xxx-L-Phe-OMe (Xxx = Aib and Ac(n)c where n = 3, 5-8) analogues were compared to determine the combined effect of backbone preferred conformation and side-chain bulkiness at position 2 on the relation of 3D-structure to biological activity. A conformational study of all the analogues was performed in solution by FT-IR absorption and 1H-NMR techniques. In parallel, each peptide was tested for its ability to induce chemotaxis, superoxide anion production and lysozyme secretion from human neutrophils. The biological and conformational data are discussed in relation to the proposed model of the chemotactic receptor on neutrophils, in particular of the hydrophobic pocket accommodating residue 2 of the tripeptide. PMID- 11860030 TI - Structural motifs in the maturation process of peptide hormones. The somatostatin precursor. I. A CD conformational study. AB - Synthetic peptides reproducing both the native domain around the dibasic cleavage site of prosomatostatin, and mutated sequences there of, previously assayed in site-directed mutagenesis experiments, have been studied by CD in different solvent systems, such as water, TFE/H2O, MeCN/H2O and aqueous SDS, in order to ascertain the ability of each solvent to stabilize secondary structural motifs. A combination of deconvolution methods and empirical calculations, that allow subtraction of the contributions due to unordered structures from the spectra, suggests that mainly two distinct families of ordered conformers containing alpha helix and/or structurally different beta-turns are present in solution, the relative stability of the different conformers depending on the nature of the solvent. The presence of beta-turns is in line with a previous NMR study in DMSO and DMSO/H2O. Comparison of the CD spectra in aqueous SDS of peptides undergoing processing with a sequence not processed in vivo shows that only the latter possesses a stable and detectable alpha-helix population. This observation suggests that the structuration involving beta-turns but no alpha-helix, which was observed by CD both in SDS and organic solvent/H2O mixtures at high water contents, might be of biological significance. The similarity of this structuration to molecular models obtained from NMR data in DMSO and DMSO/H2O is discussed. PMID- 11860031 TI - Conformational investigations on glycosylated asparagine-oligopeptides of increasing chain length. AB - Stepwise solution syntheses of the homo-oligomers Boc-(Asn)n-NHCH3 (n = 1-5; I1 5), Boc-[[GlcNAc(Ac)3beta]Asn]n-NHCH3 (n = 1-8; II1-8), and Boc [(GlcNAcbeta)Asn]n-NHCH3 (n = 1-8; III1-8) are described. Members of the series III were obtained by deacetylation of the corresponding members of the series II. The conformational preferences of the N-protected homo-peptides of the three series were investigated by spectroscopic techniques. 1H-NMR measurements were carried out in various solvents; the CD spectra were recorded in water, aqueous SDS and TFE. The poor solubility of the oligomers of the three series prevented FT-IR measurements in solution. NMR and IR measurements indicate the existence of unordered structures containing some gamma-turns in the carbohydrate-free oligomers and the presence of beta-turns in the glycosylated oligopeptides, whether acetylated or not. The CD spectra do not indicate the presence of organized structures. The sugar moieties apparently do not have a structure inducing effect on the asparagine homo-oligomer main chain. PMID- 11860032 TI - PCR detection of Tetramicra brevifilum (Microspora) infection in turbot (Scophthalmus maximus L.) musculature. AB - This study investigated the spatial distribution of Tetramicra brevifilum spores in the musculature of infected turbot Scophthalmus maximus, with the aim of identifying the most appropriate body locations for diagnostic assays. A PCR protocol optimized for the detection of T. brevifilum spores in turbot muscle is also described. In fish showing low- and moderate-intensity infection, the spatial distribution of spores was best fitted by a negative binomial distribution, indicating a clumped spatial pattern; the negative binomial coefficient k was lower for fish with low-intensity infection, indicating a more markedly clumped pattern in these fish. In fish with high-intensity infection, the spatial distribution of spores was best fitted by the Poisson distribution, indicating a random pattern. In both low- and moderate-intensity infection, spores were present at highest density in the musculature adjoining the dorsal fins. Samples for PCR were therefore obtained from this location. PCR amplification was of the small subunit ribosomal DNA (SSUrDNA), using a pair of species-specific primers that amplify the 1250 bp product. The PCR protocol developed showed better sensitivity than microscopical techniques (detection rate by microscopy 25%, versus 42% by PCR), suggesting that it may be useful for routine screening for Tetramicra brevifilum infection in cultured turbot. PMID- 11860033 TI - Recognition and invasion of human skin by Schistosoma mansoni cercariae: the key role of L-arginine. AB - The attachment of Schistosoma mansoni cercariae to mammalian skin is specifically stimulated by L-arginine. As L-arginine is an unsuitable signal for a specific identification of mammalian skin we examined the following 5 hypotheses to explain the advantage of the cercarial sensitivity to L-arginine. (1) A Schistosoma infection lowered the arginine concentration in the serum of mice, and this could enable the cercariae to avoid attachments to already infected mice. However, the infection did not reduce the arginine concentration in the skin and the cercarial attachment responses to it. (2) Creeping cercariae showed chemotactic orientation specifically along increasing L-arginine gradients. L arginine could act as a pheromone which could guide cercariae towards common penetration sites. However, the cercarial acetabular gland contents were not attractive and they did not (in contrast to previous reports) contain much arginine. (3) Schistosomula (transformed cercariae) could use L-arginine to produce nitric oxide (NO) for blood vessel dilation during their migration in the host. However, in vitro the transformed cercariae did not convert L-arginine into citrulline and NO. (4) Schistosomula could bind L-arginine from the surrounding tissues and so escape the cellular immune attack (which needs L-arginine as the precursor of NO). However, transformed cercariae bound no more L-arginine than L serine and L-lysine. (5) Schistosomula, migrating parallel to the surface in the mammalian epidermis, are dependent on information on their position between the inner and the surface layers of the skin. In the mouse skin, they adjusted their body axis with the ventral side toward the deeper (arginine-residue rich) epidermis layers. When migrating in agar, they showed chemo-orientation toward serum, and D-glucose and L-arginine were the stimulating compounds therein. The burrowing schistosomula adjusted their body axis (as in the epidermis) with the ventral side toward the higher concentration of L-arginine and not of glucose. We argue that the sensitivity for L-arginine has its primary function in orientation within mammalian skin and in location of blood vessels. PMID- 11860034 TI - Host specificity of Trypanosoma (Herpetosoma) species: evidence that bank voles (Clethrionomys glareolus) carry only one T. (H.) evotomys 18S rRNA genotype but wood mice (Apodemus sylvaticus) carry at least two polyphyletic parasites. AB - The strongest evidence for host specificity of mammalian trypanosomes comes from parasites of the subgenus Trypanosoma (Herpetosoma). Laboratory studies have shown that T. (Herpetosoma) species will not infect an alternative host. However, this has not been demonstrated in wild populations. We screened 560 bank voles (Clethrionomys glareolus) and 148 wood mice (Apodemus sylvaticus) for trypanosomes by PCR amplification of the 18S rRNA gene. In total, 109 (19%) bank voles and 12 (8%) wood mice were infected. A HaeIII restriction site was discovered that could be used to discriminate between T. (H.) evotomys of the bank vole and T. (H.) grosi of the wood mouse. All the parasites in the bank voles were identified as T. (Herpetosoma) evotomys by RFLP-PCR. Out of the 12 wood mouse infections 10 were due to T. grosi. Two of the wood mice were infected with parasites with a novel genotype that was most similar to those of T. evotomys and T. microti of voles. Fifty-six fleas collected from the rodents were also screened for trypanosomes; 9 were infected with T. evotomys and 1 with T. grosi. One of the fleas infected with T. evotomys was collected from a wood mouse. PMID- 11860035 TI - The use of host specificity, pathogenicity, and molecular markers to differentiate between Gyrodactylus salaris Malmberg, 1957 and G. thymalli Zitnan, 1960 (Monogenea: Gyrodactylidae). AB - The validity of Gyrodactylus thymalli has been questioned, based on its morphological and genetic resemblance to G. salaris. This taxonomic problem has practical implications regarding correct diagnosis of G. salaris, which has proved to be highly pathogenic to stocks of wild Norwegian Atlantic salmon. The host specificity and pathogenicity of G. salaris and G. thymalli were experimentally tested on salmon and grayling. Both parasite species were able to infect, live and reproduce on both fish species. G. salaris was highly pathogenic for the experimental stock of salmon, while grayling mounted an effective response against this parasite. Both fish species responded to an infection with G. thymalli. The results did not support the hypothesis of conspecificity between G. thymalli and G. salaris. The ribosomal RNA gene intergenic spacer of both species was sequenced. Variation in sequence was lower than expected for different species. Variation in the sequences of tandemly repeated elements was found and may prove useful in distinguishing G. salaris and G. thymalli. PMID- 11860036 TI - Ultrastructure of Buddenbrockia identifies it as a myxozoan and verifies the bilaterian origin of the Myxozoa. AB - The phylogenetic affinities of Buddenbrockia, a nematode-like parasite of freshwater bryozoans, have remained unknown since it was first reported in the nineteenth century. The discovery of Buddenbrockia parasitic in Hyalinella punctata in Ohio and Plumatella repens in France has provided material for the first ultrastructural study of this animal. This has revealed the presence of polar capsules, diagnostic myxozoan features, in the body wall. Other features, which place Buddenbrockia firmly among tetracapsulid myxozoans in the Class Malacosporea, are the unusual morphology of the polar capsules, the absence of the external tube in capsulogenesis, the body wall with its unusual cell junctions and utilization of freshwater bryozoans as hosts. The ultrastructural study has established the triploblastic organization of Buddenbrockia by confirmation of the presence of an inner layer of cells and 4 sets of longitudinal muscles. Our studies have, thus, simultaneously revealed that Buddenbrockia is a myxozoan and that the myxozoans are derived from bilaterians. The latter conclusion resolves the ongoing controversy over the triploblastic versus diploblastic nature of the Myxozoa. Our studies also provide evidence that bryozoans are ancestral hosts for the myxozoans and that loss of triploblast features has characterized the major radiation of the better known endoparasites of fish and worms in the Class Myxosporea. PMID- 11860037 TI - Risk and promotive effects in the explanation of persistent serious delinquency in boys. AB - Risk and promotive effects were investigated as predictors of persistent serious delinquency in male participants of the Pittsburgh Youth Study (R. Loeber, D. P. Farrington, M. Stouthamer-Loeber, & W. B. van Kammen, 1998), living in different neighborhoods. Participants were studied over ages 13-19 years for the oldest sample and 7-13 years for the youngest sample. Risk and promotive effects were studied in 6 domains: child behavior, child attitudes, school and leisure activities, peer behaviors, family functioning, and demographics. Regression models improved when promotive effects were included with risk effects in predicting persistent serious delinquency. Disadvantaged neighborhoods, compared with better neighborhoods, had a higher prevalence of risk effects and a lower prevalence of promotive effects. However, predictive relations between risk and promotive effects and persistent serious delinquency were linear and similar across neighborhood socioeconomic status. PMID- 11860038 TI - Clinical adolescent psychology: what it is, and what it needs to be. AB - This commentary on the special section on clinical adolescent psychology (G. Holmbeck & P. Kendall. 2002) reviews and critiques the conceptual and empirical articles that this compilation comprises. As articulated in the conceptual contributions to this collection, two fundamental principles should guide research on the etiology, prevention, and treatment of psychological disorder and dysfunction during adolescence: First, drawing on the fiel of developmental psychopathology, the study of clinical adolescent psychology should focus on the trajectories of disorder that precede, characterize, and follow adolescence. Second, drawing on the literature on normative adolescent development, the study of clinical adolescent psychology must proceed with an explicit recognition of the unique biological, cognitive, psychosocial, and contextual features that define adolescence as a developmental period. The empirical contributions to this compilation are evaluated with respect to the extent to which they reflect these tenets. Although the study of clinical adolescent psychology, as evidenced by this collection of articles, is appropriately grounded in the broader enterprise of developmental psychopathology, less progress has been made with respect to the integration of the study of clinical phenomena in adolescence with the study of normative adolescent development. PMID- 11860039 TI - Depressive symptoms and close relationships during the transition to adulthood: perspectives from dysphoric women, their best friends, and their romantic partners. AB - Late adolescent women's depressive symptoms and interpersonal functioning were assessed using reports from participants, their best friends, and their romantic partners. As predicted, the associations between relationship dysfunction and dysphoria were stronger in romantic relationships than in friendships. Unlike friends, romantic partners perceived dysphoric women as having poorer social skills. Romantic partners also reported providing less emotional support to dysphoric women, whereas friends reported providing more. Finally, romantic partners of dysphoric women had more Cluster A (odd-eccentric) personality disorder symptoms; these symptoms mediated the relation between women's depression and partners' nonsupportiveness. The findings suggest that dysphoric women may find themselves in emotionally nonsupportive romantic relationships because they have paired (through assortative pairing or mutual influence) with symptomatic partners. PMID- 11860040 TI - Mood disturbance fails to resolve across 31 days of cigarette abstinence in women. AB - Smoking abstinence responses were characterized in 96 female smokers. Participants completed subjective state measures twice per week for 5 weeks and were then randomly assigned to a group required to abstain for 31 days or a control group that continued to smoke. Financial incentives for biochemically verified abstinence resulted in an 81% completion rate. Abstinence-related increases in depression, tension, anger, irritability, and appetite showed little tendency to return to prequit levels and remained significantly elevated above smoke-group levels. In contrast to psychological components of anxiety, physical components decreased to smoke group levels by the 2nd week of abstinence. Trait depression and neuroticism predicted larger increased abstinence-associated negative affect. The Big Five personality dimensions predicted variance not associated with depressive traits. PMID- 11860041 TI - Inhibitory processes in adults with persistent childhood onset ADHD. AB - The theory that attention-deficit/hyperactivity disorder (ADHD) stems from a deficit in an executive behavioral inhibition process has been little studied in adults, where the validity of ADHD is in debate. This study examined, in high functioning young adults with persistent ADHD and a control group, 2 leading measures of inhibitory control: the antisaccad task and the negative priming task. ADHD adults showed weakened ability to effortfully stop a refle ve or anticipated oculomotor response but had normal ability to automatically suppress irrelevant information. Results suggest that an inhibitory deficit in ADHD is confined to effortful inhibition of motor response, that antisaccade and negative priming tasks index distinct inhibition systems, and that persistence of ADHD symptoms into adulthood is associated with persistence of executive motor inhibition deficits. PMID- 11860042 TI - Assessing match and mismatch between practitioner-generated and standardized interview-generated diagnoses for clinic-referred children and adolescents. AB - Although the Diagnostic and Statistical Manual of Mental Disorders (DSM) is widely used in both clinical and research settings, little is known about agreement between clinician and standardized research diagnoses. Diagnoses generated by the Diagnostic Interview Schedule for Children (DISC-P-2.3) were compared with clinician-generated diagnoses for 245 referred youths. Agreement was poor for all individual disorders and broader diagnostic clusters. Agreement was higher for externalizing categories than for internalizing, but no association was found between agreement and child, family, or clinician characteristics. Clinicians were more likely than the DISC to assign 1 diagnosis and less likely to assign 0 diagnoses, suggesting that clinic policies may play a role. Implications for the use of the DSM across different settings are discussed. PMID- 11860043 TI - Does disclosure of emotions facilitate recovery from bereavement? Evidence from two prospective studies. AB - Two longitudinal studies assessed whether disclosure of emotions facilitates recovery from bereavement. Study 1 tested prospectively over a 2-year period whether the extent to which bereaved persons talked about their loss to others and disclosed their emotions was associated with better adjustment to the loss of a marital partner. There was no evidence that disclosure facilitated adjustment. Study 2 randomly assigned recently bereaved individuals either to the Pennebaker writing task (J. W. Pennebaker & S. K. Beall, 1986) or to no-essay control conditions. The writing task did not result in a reduction of distress or of doctors visits either immediately after the bereavement or at a 6-month follow up. Beneficial effects were not demonstrated for bereaved persons who had suffered an unexpected loss or who at the time of the study still expressed a high need for emotional disclosure. PMID- 11860044 TI - A cognitive-ecological approach to preventing aggression in urban settings: initial outcomes for high-risk children. AB - A multiyear, multicontext aggression prevention intervention was provided during the early or late elementary school years in an inner-city and an urban poor community. Sixteen schools were randomly assigned to 1 of 4 conditions: (a) no treatment control, (b) general enhancement classroom program, (c) general enhancement plus small-group peer-skills training, or (d) general enhancement plus small-group peer-skills training plus family intervention. This article reports on results for the high-risk subsample of 1,500 children. Results from hierarchical linear modeling indicate that comprehensive interventions, if provided in early grades, can be effective for children in schools in settings with resources adequate to support learning and development, but some unintended effects can occur in schools in the most distressed communities when delivered too late in development. PMID- 11860045 TI - Expectant parents' representations of early attachment relationships: associations with mental health and family history. AB - The association between adult representations of early attachment relationships and history of individual and family mental health was examined in a sample of 233 expectant mothers and fathers. As predicted, security of attachment was linked to mental health. Parents classified as Preoccupied were more likely than other parents to report suicidal ideation. Whereas parents classified as Unresolved more often reported suicidal ideation, emotional distress, and substance abuse. With respect to family history. Unresolved and Preoccupied attachment classifications were significantly related to child abuse involving a relative and parental separation or divorce. These findings support theoretical conceptualizations regarding the link between adult attachment and mental health in middle-class American adults. PMID- 11860046 TI - Posttraumatic stress symptoms and victimization among Japanese American women. AB - This study of Japanese American women and immigrant women from Japan investigated the relationship between posttraumatic stress (PTS) symptoms and the perceived abusiveness of partners' emotional and physical violence, with a community-based randon sample. Women who experienced injuries and/or fear for their lives, in addition to partners' emotional and physical violence, had significantly higher PTS symptom counts than those with no lifetime experience of partners' violence. Victimization by nonintimates also increased PTS symptom counts. Satisfaction with social support significantly mitigated the negative effect of childhood abuse for reexperiencing and avoidance symptoms. PMID- 11860047 TI - Developmental factors in the treatment of adolescents. AB - Most empirically supported interventions for adolescent mental health problems are either downward adaptations of adult treatments or upward adaptations of child treatments. Although these treatments show respectable effects with teens, a review of the outcome research reveals significant gaps. both in coverage of adolescent conditions and problems (e.g., eating disorders, suicidality) and in attention to the biological, psychological, and social dimensions of adolescent development. The authors critique the field, propose a biopsychosocial framework for the development of dysfunction and intervention, and discuss ways the developmental literature can and cannot inform intervention and research. A long term goal is an array of developmentally tailored treatments that are effective with clinically referred teens and an enriched understanding of when, how, and why the treatments work. PMID- 11860048 TI - Predicting relapse back to smoking: contrasting affective and physical models of dependence. AB - Traditional models of physical dependence suggest that nicotine dependence should be reflected by the extent of drug exposure (e.g., smoking rate) and by evidence of physiological adaptation (e.g., withdrawal severity). An affective model suggests that nicotine dependence should be related to an individual's tendency to experience negative affect and expectations that nicotine use would ameliorate such affect. This research investigated the ability of these 2 models to predict relapse back to smoking at 6 months postquit. Logistic regression models were developed and tested in 505 heavy smokers participating in nicotine patch clinical trials. Results supported both models, but the most potent predictor of outcome was postquit negative affect, which accounted for much of the predictive validity of traditional measures of nicotine dependence. Affective reactivity appears to be a core constituent of dependence. PMID- 11860049 TI - Magnitude and duration of cardiovascular responses to anger in Vietnam veterans with and without posttraumatic stress disorder. AB - This study investigated the cardiovascular responses to a relived anger task in 118 male Vietnam combat veterans (62 with posttraumatic stress disorder [PTSD] and 56 without PTSD). Participants completed standardized diagnostic measures, hostility measures, and a laboratory session in which they relived a self-chosen anger memory while heart rate (HR), systolic blood pressure, and diastolic blood pressure (DBP) were measured continuously using an Ohmeda Finapres monitor. Compared with veterans without PTSD, PTSD veterans took less time to feel anger, had greater mean HR and DBP response during relived anger, and reported greater anger and anxiety during the task. There was a significant relationship between covert hostility and anger response, during and after the anger task only in participants with PTSD. PMID- 11860050 TI - Predictors of posttraumatic stress disorder symptomatology following bone marrow transplantation for cancer. AB - This study examined the prevalence and predictors of posttraumatic stress disorder (PTSD) symptoms in 70 men and women treated with bone marrow transplantation for cancer. Findings indicated that the number of symptoms present ranged from 0 to a possible high of 17 (M = 3.0, SD = 3.9). As predicted. lower social support and higher avoidance coping I month pretransplant predicted greater PTSD symptom severity an average of 7 months posttransplant. These variables remained significant predictors of symptom severity even after accounting for pretransplant levels of psychological distress. Additional analyses indicated the presence of a significant interaction between social support and avoidance coping, with patients high in avoidance coping and low in social support reporting the most severe symptoms. These findings identify patients at risk for psychological disturbance posttransplant and can serve to guide future intervention efforts. PMID- 11860051 TI - Trauma exposure and posttraumatic stress disorder in psychosis: findings from a first-admission cohort. AB - This study examined the lifetime prevalence of trauma exposure and posttraumatic stress disorder (PTSD) and their demographic, diagnostic, and trauma-related correlates in a clinical cohort of 426 patients with a first psychiatric admission for psychosis. The prevalence of trauma exposure was 68.5%. Female gender and substance abuse were risk factors for trauma exposure. The prevalence of PTSD was 14.3% in the full sample and 26.5% in those with trauma exposure. PTSD was less prevalent in patients with bipolar disorder and schizophrenia and was twice as common in women. Other significant risk factors were younger age and trauma exposure that was repeated and ongoing or that involved childhood victimization. The findings highlight the importance of systematically ascertaining trauma histories in patients with psychotic disorders. PMID- 11860052 TI - The impact of perceived child physical and sexual abuse history on Native American women's psychological well-being and AIDS risk. AB - The impact of perceived child abuse history on 160 adult, Native American women's emotional well-being (i.e., depressive mood and anger) and AIDS risk was examined. How sense of mastery and social support might lead to women's greater resiliency was also investigated. Child physical-emotional abuse was found to have greater impact on depressive mood and anger and AIDS risk than did child sexual abuse. This finding was independent of current stress in women's lives. Women who were physically-emotionally abused as children had 5.14 times greater odds of having a sexually transmitted disease in their lifetimes than did women who experienced only marginal or no physical-emotional abuse. Moreover, consistent with the communal culture of Native Americans, social support was found to contribute more to resilience than sense mastery did. Reasons for the greater predictive power of child physical-emotional abuse compared with child sexual abuse in a growing number of studies are discussed. PMID- 11860053 TI - Why not to screen high-risk women anticipating BRCA1/BRCA2 testing for psychological distress. PMID- 11860054 TI - Introduction to the special section on clinical adolescent psychology: developmental psychopathology and treatment. AB - This article addresses implications of the interface between developmental psychology and clinical psychology for research on adolescence and describes the importance of considering developmental level when designing treatments for adolescent patients. In addition, the articles that constitute the special section, "Clinical Adolescent Psychology: Developmental Psychopathology and Treatment," are introduced. PMID- 11860055 TI - Cultural and contextual influences in mental health help seeking: a focus on ethnic minority youth. AB - In this article, a mental health help-seeking model is offered as a framework for understanding cultural and contextual factors that affect ethnic minority adolescents' pathways into mental health services. The effects of culture and context are profound across the entire help-seeking pathway, from problem identification to choice of treatment providers. The authors argue that an understanding of these help-seeking pathways provides insights into ethnic group differences in mental health care utilization and that further research in this area is needed. PMID- 11860056 TI - Attachment and autonomy as predictors of the development of social skills and delinquency during midadolescence. AB - This study examined adolescent attachment organization as a predictor of the development of social skills and delinquent behavior during midadolescence. Delinquent activity and skill levels were assessed for 117 moderately at-risk adolescents at ages 16 and 18, and maternal and adolescent attachment organization and autonomy in interactions were assessed at age 16. Adolescent attachment security predicted relative increases in social skills from age 16 to 18, whereas an insecure-preoccupied attachment organization predicted increasing delinquency during this period. In addition, preoccupied teens interacting with highly autonomous mothers showed greater relative decreases in skill levels and increases in delinquent activity over time, suggesting a heightened risk for deviance among preoccupied teens who may be threatened by growing autonomy in adolescent-parent interactions. PMID- 11860057 TI - A developmental psychopathology perspective on adolescence. AB - Developmental psychopathology offers an integrative framework for conceptualizing the course of development during adolescence, with particular relevance for understanding continuity and the emergence of psychopathology during this and subsequent developmental periods. In this article, the utility of a developmental psychopathology perspective for informing the design of research, prevention, and intervention is highlighted. Interdisciplinary, organizational models of development, emphasizing the dynamic relations between the developing individual and internal and external contexts, are discussed. Examination of boundaries between abnormal and normal development during adolescence offers important vantage points for articulating diversity in the developmental course during this period. Conceptualizing divergence and convergence in developmental pathways, continuity and discontinuity in development, and the transactions of risk and protective processes leading to maladaptation, psychopathology, and resilience are highlighted. PMID- 11860058 TI - Binge drinking trajectories from adolescence to emerging adulthood in a high-risk sample: predictors and substance abuse outcomes. AB - This study describes binge drinking trajectories from adolescence to emerging adulthood in 238 children of alcoholics and 208 controls. Mixture modeling identified three trajectory groups: early-heavy (early onset, high frequency), late-moderate (later onset, moderate frequency), and infrequent (early onset, low frequency). Nonbingers were defined a priori. The early-heavy group was characterized by parental alcoholism and antisociality, peer drinking, drug use, and (for boys) high levels of externalizing behavior, but low depression. The infrequent group was elevated in parent alcoholism and (for girls) adolescent depression, whereas the nonbinger and late-moderate groups showed the most favorable adolescent psychosocial variables. All 3 drinking trajectory groups raised risk for later substance abuse or dependence compared with the nonbingers, with the early-heavy group at highest risk. PMID- 11860059 TI - Developmental trajectories of adolescents' depressive symptoms: predictors of change. AB - In a sample of 240 adolescents assessed annually in Grades 6 through 11, the developmental trajectories of their depressive symptoms were examined using latent factor growth modeling. Growth in mother-reported adolescent depressive symptoms was quadratic; growth in adolescent-reported symptoms was linear. In the model with gender and maternal depression, girls reported a greater increase in depressive symptoms over time than boys, and adolescents of mothers with histories of mood disorders had higher initial levels of depressive symptoms than offspring of never-depressed mothers. After gender and maternal depression were controlled, initial levels of negative attributions and stressors significantly predicted initial levels of adolescent- and mother-reported depressive symptoms. Attributional styles that were increasingly negative across time were associated with significantly higher initial levels (mother reported) and increasing growth (adolescent reported) of depressive symptoms. Reciprocal models in which development of depressive symptoms predicted the development of attributions and stress also were examined. PMID- 11860060 TI - Observed and perceived parental overprotection in relation to psychosocial adjustment in preadolescents with a physical disability: the mediational role of behavioral autonomy. AB - The purpose of this study was to tes a mediational model of associations between parental overprotectiveness (OP), behavioral autonomy. and psychosocial adjustment in 68 families with 8- and 9-year-old preadolescents with spipa bifida and a demographically matched sample of 68 families with able-bodied children. Measures included questionnaire and observational assessments of parental OP; parent and child reports of behavioral autonomy; and parent, child, and teacher reports of preadolescent adjustment. On the basis of both questionnaire and observational measures of OP, mothers and fathers of children with spina bifida were significantly more overprotective than their counterparts in the able-bodied sample, although this group difference was partially mediated by children's cognitive ability. Across samples, mothers were more likely to be overprotective than fathers. Both questionnaire and observational measures of parental OP were associated with lower levels of preadolescent decision-making autonomy as well as with parents being less willing to grant autonomy to their offspring in the future. For the questionnaire measure of OP, and only for the spina bifida sample. the mediational model was supported such that parental OP was associated with less behavioral autonomy, which was, in turn, associated with more externalizing problems. Findings are discussed in relation to the literature on parenting, autonomy development, and pediatric psychology. PMID- 11860061 TI - The three faces of vestibular ganglionitis. AB - We present temporal bone and clinical evidence that common syndromes of recurrent vertigo are caused by a viral infection of the vestibular ganglion. In the present series, histopathologic and radiologic changes in the vestibular ganglion and meatal ganglion were consistent with a viral inflammation of ganglion cells in cases of Meniere's disease, benign paroxysmal positional vertigo, and vestibular neuronitis. Clinical observations of multiple neuropathies involving cranial nerves V, VII, and VIII on the same side in patients with recurrent vertigo are best explained by a cranial polyganglionitis caused by a neurotrophic virus, which is reactivated by a stressful event later in life. The reactivation of the latent virus may manifest as one of the above vertigo syndromes, depending on the part of the vestibular ganglion that is inflamed, the type and strain of the virus, and host resistance. PMID- 11860062 TI - Sudden death after neck dissection for cancer. AB - The goal of this study was to analyze the mortality data following neck dissection and determine the risk factors of early death. The hospital mortality records were analyzed from 3,015 consecutive patients who underwent neck dissection. A case control study analyzed risk factors of death during the first 3 postoperative days. The mortality incidences were 0.50% and 1.33%, respectively, during the first 3 and the first 30 postoperative days. Eleven of the 12 unexplained deaths occurred during the first 3 postoperative days, and most of these patients died suddenly. They were more likely to be alcoholic and to have undergone nerve section. In most of the patients who died after the third postoperative day, death was related to a postoperative complication. Although the mechanisms of sudden death remain unclear, careful follow-up of these patients during the early postoperative days should be performed to reduce the mortality risk by shortening the delay of care. PMID- 11860064 TI - Waveform changes in antidromic facial nerve responses in patients with Bell's palsy. AB - We repeatedly tested the antidromic facial nerve response within 7 days after onset of paralysis in patients with Bell's palsy. None of 109 patients showed the triphasic waveform that reflects normal conduction of the facial nerve action potential. The waves recorded from patients showed biphasic, monophasic, or flat waveforms. Eighty-two of 88 patients with complete recovery showed biphasic waves, whereas half of the patients with nerve degeneration had monophasic or flat waves. Most patients with complete recovery maintained biphasic waves, but in patients with incomplete recovery, the waveforms changed to monophasic or flat, except in 1 case. The presence of monophasic or flat waves with a low facial score strongly suggests nerve degeneration. The antidromic facial nerve response is recommended as a method of diagnosing paralysis and monitoring the progression of intratemporal facial nerve damage during its early stages. PMID- 11860065 TI - Antimicrobial defensin peptides of the human nasal mucosa. AB - Defensins, a prominent group of antimicrobial peptides, are an important component of the innate immune response, particularly at mucosal surfaces that are vulnerable to colonization by potential pathogens. The present study was undertaken to investigate the expression of defensins in inferior turbinate mucosa of normal subjects and inferior turbinate mucosa and nasal polyps of patients with chronic sinusitis. Expression of beta-defensin 1 and 2 and alpha defensin 5 and 6 messenger RNAs (mRNAs) was investigated by reverse transcriptase polymerase chain reaction, and their expression level was semiquantitatively evaluated by dot blot hybridization. Immunohistochemical analysis was used for detection of alpha-defensins 1, 2, and 3 in tissue sections. Beta-defensin 1 mRNA was expressed in all tissue samples, at levels that did not differ significantly. Beta-defensin 2 mRNA was detected in the turbinate mucosa and nasal polyps of patients with chronic sinusitis, but not in normal mucosa. Its expression level was significantly higher in nasal polyps than in turbinate mucosa. Alpha-defensin 5 and 6 mRNAs were not expressed in any tissues, but alpha-defensins 1, 2, and 3 were detected in all tissue samples obtained from patients with chronic sinusitis. These results suggest that beta-defensin 1 may play a constitutive role in nasal defenses, whereas alpha-defensins 1, 2, and 3 and beta-defensin 2 may be induced in response to local infection or inflammation. PMID- 11860063 TI - Unloaded shortening velocity and myosin heavy chain variations in human laryngeal muscle fibers. AB - Myosin description in human laryngeal muscles is incomplete, but evidence suggests the presence of type I, IIA, IIX, and tonic myosin heavy chain (MHC) fibers. This study describes the unloaded shortening velocity (V0) of chemically skinned laryngeal muscle fibers measured by the slack test method in relation to MHC content. Skeletal fibers from human laryngeal and limb muscle biopsy specimens were obtained for determination of V0, and subsequently, glycerol sodium dodecyl sulfate-polyacrylamide gel electrophoresis was used to determine the MHC isoform content. The fibers from human limb muscle had shortening speeds similar to those in previous reports on human skeletal fibers. Type I, IIA, and IIX fibers of laryngeal muscle had shortening speeds similar to those of fibers from limb muscle, but laryngeal fibers with heterogeneous MHC expression had a wide range of shortening speeds, some being nearly twice as fast as limb fibers. In addition, MHC isoform bands from human extraocular muscle comigrated with some bands from laryngeal muscle--a finding suggesting that extraocular myosin may also be expressed. PMID- 11860066 TI - In vivo measurements of an improved tracheostoma valve based on inhalation. AB - An improved, inhalation-based tracheostoma valve (TSV) was designed to facilitate finger-free tracheoesophageal speech. In contrast to existing models, the TSV closes by means of strong inhalation (instead of exhalation) to reach the "speaking position." Air is inhaled through a small 1-way valve that allows unlimited phonation time. The device can be deliberately changed to the "breathing position" by a fast expiration. Experimental setups were used to measure in vitro and in vivo the performance of the inhalation TSV. In patients, the inhalation TSV was compared with existing TSVs on the following parameters: air volume used to close the TSVs, speaking time of the TSVs, and total air volume during exhalation through the TSVs. The inhalation TSV functions well in physiological ranges, is optimally adjustable, and is an improvement over existing devices. It makes continuous speech possible and saves as much as 22% of the total exhalation volume for speaking, in contrast to existing TSVs. PMID- 11860067 TI - Detection of cytokeratins in normal and malignant laryngeal epithelia by means of reverse transcriptase-polymerase chain reaction. AB - Cytokeratins (CKs) are a subgroup of intermediate filament proteins that take part in forming the cytoskeleton. The epithelial cells in different organs express distinct CKs, and this expression may be modified during malignant transformation. Here we describe the use of reverse transcriptase-polymerase chain reaction followed by Southern blotting to determine the profile of expression of CKs in both normal and malignant laryngeal samples. Thirty-six samples were subjected to histologic examination and classified as 17 squamous cell carcinomas, 3 dysplastic lesions, and 16 normal samples. CK8 and CK19 were expressed in almost all samples, both cancerous and normal, and were therefore used to verify the integrity of RNA. Expression of CK2, CK9, and CK20 was not detected in any of the samples, normal or cancerous. CK15 and CK18 showed low sensitivity for detection of cancer (36.4% and 45.5%, respectively). CK10 showed relatively high sensitivity (91%), but only moderate specificity (69.2%). Only CK17 showed both high sensitivity and specificity (91% and 92.3%, respectively; positive predictive value, 91%). We propose that CK17 may be considered a promising candidate to use as a molecular marker for malignant transformation in laryngeal squamous cell carcinoma. PMID- 11860068 TI - Revision functional endoscopic sinus surgery. AB - Functional endoscopic sinus surgery (FESS) has recently become a popular procedure for treating chronic sinusitis. However, there is a 2% to 24% rate of primary FESS failure. Revision functional endoscopic sinus surgery (RESS) is indicated for patients who remain symptomatic after primary FESS and optimal medical therapy have failed. A retrospective study was conducted on all patients who underwent RESS in our department between April 1988 and March 1998. During this period, FESS was performed 1,227 times to treat chronic sinusitis; among those procedures, 142 were RESS. Complications occurred in 14 RESS procedures (9.9%). After RESS, 65% patients had improved. In this study, we concluded that although RESS is more difficult than primary FESS because of distorted or missing anatomic landmarks, complications did not increase with RESS if meticulous surgical procedures were performed. However, the 65% improvement rate with RESS was lower than that of primary FESS. PMID- 11860069 TI - Clinical study of acute low-tone sensorineural hearing loss: survey and analysis of glycerol test and orthostatic test. AB - Fifty patients (14 men and 36 women ranging in age from 16 to 66 years) with acute low-tone sensorineural hearing loss (ALHL) were retrospectively analyzed in this study. The glycerol test and the orthostatic test were performed. On the glycerol test, 43.8% of 16 ears of the 14 men and 33.3% of 39 ears of the 36 women with ALHL had a positive result. On the orthostatic test, 42.9% of the 14 men and 52.7% of the 36 women with ALHL had a positive result. In addition, 28.6% of the 14 men and 33.3% of the 36 women had hypotension with a systolic blood pressure of 100 mm Hg or less. Our results suggest that endolymphatic hydrops as shown by the glycerol test and an underlying autonomic imbalance and/or insufficient blood circulation as shown by the orthostatic test may be important factors in the causation of ALHL. PMID- 11860070 TI - Acute tonsillitis as the first manifestation of post-transplant lymphoproliferative disorder. AB - Post-transplant lymphoproliferative disorder (PTLD) is a life-threatening complication that may follow orthotopic liver transplantation (OLT) in children. The first symptoms are often in the ear, nose, or throat (ENT) area. This abnormal proliferation of lymphoid cells is related to Epstein-Barr virus (EBV) infection in immunocompromised children. The incidence of PTLD, EBV status before OLT and at the diagnosis of PTLD, delay between OLT and PTLD, localization, pathological investigations, and the treatment and evolution of PTLD were prospectively evaluated in 77 pediatric liver transplant recipients. Eight patients (10%) developed PTLD, all with an ENT presentation. Seven had acute nonbacterial tonsillitis (with a negative throat swab), and 1 had a pharyngolaryngeal localization at the time of the diagnosis. Four patients had associated involvement outside the ENT area. All patients were EBV-seronegative at the time of OLT; 6 underwent seroconversion at the time of diagnosis, and 2 within 9 and 20 months of diagnosis. All patients presented with low-grade PTLD. All patients with acute tonsillitis associated with EBV seroconversion underwent immediate tonsillectomy, and immunosuppression was decreased as much as tolerated. This therapeutic protocol led to complete recovery in all patients. After OLT in children, nonbacterial tonsillar inflammation or hypertrophy associated with an EBV infection is often the first manifestation of PTLD. Tonsillectomy combined with tapering of immunosuppression offers the best chance for a complete recovery. PMID- 11860071 TI - Functional neck dissection for the clinically negative neck: effectiveness and controversies. AB - This study was performed to evaluate the effectiveness of functional neck dissection in controlling metastasis to the clinically negative (cN0) neck, focusing on recurrences in the pathologically negative (pN0) neck and the role of extracapsular spread in the cN0 neck. A series of 172 patients (253 dissected fields) treated for cN0 laryngeal or hypopharyngeal cancer with a 5-year minimum follow-up is presented. Occult metastasis was observed in 30% of the patients. Extracapsular spread was present in 39% of the positive nodes. The neck recurrence rate was 5.2%. Surgical specimens from cases of neck recurrence in pN0 necks were reevaluated for micrometastasis by immunostaining with antibody for cytokeratins. The immunohistochemical findings were positive in 1 of 4 cases. Functional neck dissection provides good neck control and survival rates for the cN0 neck. The accurate prognostic significance of extracapsular spread in cN0 necks is still unknown. Micrometastasis alone may be insufficient to explain recurrences in pN0 necks. PMID- 11860072 TI - Complications of intranasal prescription narcotic abuse. AB - The abuse of drugs via an intranasal route is an increasingly prevalent pattern of behavior. In the past year, a number of patients received care at our institution for complications resulting from the previously unreported phenomenon of intranasal prescription narcotic abuse. This report describes the clinical manifestations of this form of drug abuse in 5 patients. Their symptoms consisted of nasal and/or facial pain, nasal obstruction, and chronic foul-smelling drainage. Common physical findings were nasal septal perforation; erosion of the lateral nasal walls, nasopharynx, and soft palate; and mucopurulent exudate on affected nasal surfaces. In addition, 2 of the 5 patients had invasive fungal rhinosinusitis, which appears to be a complication unique to intranasal narcotic abuse. PMID- 11860073 TI - Gastroesophagopharyngeal reflux in patients with contact granuloma: a prospective controlled study. AB - The aim of this study was to compare the incidences of gastroesophagopharyngeal reflux in patients with contact granuloma and healthy controls. A 24-hour ambulatory esophagopharyngeal pH monitoring technique was used to measure reflux parameters in the pharynx and distal esophagus. Pharyngeal acid reflux events occurred in 17 of 26 granuloma patients (1 to 20 episodes per patient) and 5 of 19 controls (1 to 8 episodes per patient). The reflux episodes were typically short and occurred predominantly in an upright position. A comparison between the groups showed a significant difference in the number of pharyngeal reflux episodes (p = .009) and in the total time of pH below 4 (p = .006). On the other hand, we found no significant differences in any esophageal reflux parameters, except for the percentage of distal esophageal reflux episodes that reached the pharynx (p = .006). In this study, pharyngeal acid exposure was significantly more prevalent in patients with contact granuloma than in healthy controls. PMID- 11860074 TI - Similar COL1A1 expression in fibroblasts from some patients with clinical otosclerosis and those with type I osteogenesis imperfecta. AB - Because of the clinical and histopathologic similarities between otosclerosis and type I osteogenesis imperfecta, we examined COL1A1 messenger RNA (mRNA) expression in cultured fibroblasts from patients with clinical otosclerosis to determine whether abnormalities of expression of COL1A1 were present, as has been reported in type I osteogenesis imperfecta. Type I osteogenesis imperfecta has been found to result from mutations in the COL1A1 gene that result in null expression of the mutant allele. Patients with clinical otosclerosis were genotyped for the presence of an expressed 4 base-pair insertion polymorphism in the 3' region of the COL1A1 gene. Skin biopsies were performed, and cultured fibroblast cell lines were established from patients who were heterozygous for the polymorphism. Allelic expression was examined by reverse transcription polymerase chain reaction and silver-stained polyacrylamide gel electrophoresis. Two of 9 patients with clinical otosclerosis demonstrated null or reduced expression of one COL1A1 allele. The differential expression of the two COL1A1 alleles in all subjects was also examined by a semiquantitative method using an ABI Prism 7700 Sequence Detection System (Taqman). We did this examination to determine whether milder abnormalities in COL1A1 expression might account for the development of otosclerosis in the 7 clinical cases that did not reveal evidence of null expression by the gel technique. Of the same 2 cases of otosclerosis that demonstrated evidence of null expression by gel electrophoresis, both were found to have significant differences in COL1A1 mRNA expression by the Taqman analysis. The remaining 7 cases revealed equal expression of the two COL1A1 alleles similar to that seen in controls. These results suggest that mutations in COL1A1 that are similar to those that occur in type I osteogenesis imperfecta may account for a small percentage of cases of otosclerosis, and that the majority of cases of clinical otosclerosis are related to other genetic abnormalities that have yet to be identified. PMID- 11860075 TI - Simple on-line endotracheal cuff pressure relief valve. AB - Ischemic injury of the tracheal mucosa in the endotracheally intubated patient is directly proportional to the tracheal tube cuff pressure. At a cuff pressure of 30 cm H2O, the tracheal mucosal blood flow becomes partially obstructed, and at a pressure of 45 cm H2O, the obstruction to the tracheal mucosal blood becomes total, leading to tracheal mucosal damage and subsequent complications. In our institute, we have developed a simple and very inexpensive method to gauge the cuff pressure. We use a regular 20-mL syringe attached in line with the connector of the endotracheal tube cuff. In this manner, we monitored the intracuff pressure in 120 patients who underwent ear or neck surgery. The syringe was connected to the tube cuff and inflated with 15 mL of air. The syringe was left constantly connected to the cuff. In addition, the cuff pressure was measured with the Mallinckrodt Hi-Lo aneroid pressure gauge at the beginning of surgery and hourly thereafter for the duration of surgery. At the same time, a check for leakage around the cuff was made by auscultation with a stethoscope above the sternal notch. Multiple comparisons between the repetitive intracuff pressure measurements revealed that there were no significant differences in the intracuff pressure values measured at the different times of surgery. These results indicate that there was an adequate venting of the excess intracuff pressure and also that there was no leakage around the cuff. PMID- 11860076 TI - Glossopharyngeal neuralgia associated with aberrant internal carotid artery in the oropharynx. PMID- 11860077 TI - Cost shifting revisited: the case of service intensity. AB - This paper examines whether a health care provider's choice of service intensity for any patient group affects its cost shifting behavior. Our theoretical models indicate that firms may respond to lower prospective payment by decreasing service intensity to all of its patient groups, thereby giving firms an alternative to cost shifting. Additionally, the conditions under which cost shifting and lower service intensity occur are identical, regardless of profit status. Using a panel of California hospitals, we found that nonprofit hospitals do cost shift, while profit-maximizing hospitals do not. However, both firms respond to lower prospective payment by decreasing service intensity, thus supporting our theoretical conclusion that lower service intensity can be used as an alternative to cost shifting. PMID- 11860078 TI - Buying healthcare quality with grants and donations. AB - We find that grants and contracts increase California healthcare clinics' average costs, but donations do not. We interpret these results as meaning that grants and contracts change the nature of the product while donations are used to increase output without changing the product. One possible scenario is that grants and contracts are targeted money used to enhance quality. A theoretical analysis shows that quality is enhanced more if grants and contracts are used to reward quality rather than as seed money to create quality. The empirical evidence, however, is that grantors are motivated by the second reason. PMID- 11860079 TI - An econometric analysis of screening and treatment of patients with suspected Chlamydia. AB - Chlamydia trachomatis is probably the most common sexually transmitted disease in the Western industrialised countries with devastating consequences. However, it is an infection that can be so easily treated. There are over 50 million new cases occurring each year. In the United States chlamydia is seen as the most common and costly of the bacterial sexually transmitted diseases (STD), with approximately 4 million new cases occurring each year at an estimated total cost of $2.4 billion. The characteristic of this infection is its difficulty of detection that promotes its spread and making its prediction rather complex. Chlamydial infections are commonly asymptomatic or cause mild or non-specific symptoms and signs, which are not easily detected. Approximately 70% of women with endocervical infections and up to 50% of men with urethral infections are asymptomatic and thus not likely to seek medical care. Chlamydia has become known as the "silent epidemic". It is the more frequently identifiable single cause of pelvic inflammatory disease (PID), occurring in an estimated 15-40% of women. The primary objective of the study was to identify factors and quantify their contribution to the risk of being infected with Chlamydia and to construct an easy to use friendly method for early detection. The importance of developing some means of early detection is vital and previous studies suggest that selective screening might be one solution. A logit model was fitted to three broad variables: behavioural, patients' characteristics, and signs/symptoms noted by patient. The age of the women, the number of sexual partners over the past year, previous history of sexually transmitted disease, the use of barrier contraception and patients' and their partners' signs and symptoms were found to be among the most important variables. Such a model should allow patients who are in a high-risk category, allowing appropriate treatment. PMID- 11860080 TI - Improving the sensitivity of the time trade-off method: results of an experiment using chained TTO questions. AB - We test a chained time trade-off (TTO) approach to estimating health gains from interventions by asking respondents to directly compare the "before" and "after" intervention health states in the TTO framework. Respondents with experience of both health states were used, thus minimising biases stemming from confusion surrounding health descriptions. We found that responses to these direct comparisons were much more likely to capture a perceived change in health status than the conventional approach to TTO estimation. This is an important finding because the TTO method is preferred by practitioners to many other direct generic methods of health status valuation both on empirical grounds and because it is based on the notion of opportunity cost, which is central to consumer theory, requiring respondents to express their preferences in terms of foregoing some of one good in exchange for more of another (unlike a rating scale). PMID- 11860081 TI - Measuring hospital efficiency in Austria--a DEA approach. AB - We investigate the evolution of efficiency and productivity in the hospital sector of an Austrian province for the time period 1994-1996. We use panel data to design non-parametric frontier models (Data Envelopment Analysis) and compare efficiency scores and time patterns of efficiency across medical fields. As health outcomes hardly can be measured in a direct way we make use of two different approaches for output measurement: In a first approach, we employ the number of case mix-adjusted discharges and of inpatient days, in a second we use credit points, which are calculated in course of the newly introduced diagnosis related group-type financing system. We calculate and compare individual efficiency scores for hospital wards as decision making units (DMU) in specified medical fields. To our knowledge the calculation of ward-specific efficiency scores has not up till now been the unit of non-parametric efficiency analysis. Our two models find different results: Model 1 with conservative output measurement calculates an average efficiency level of 96%, while model 2 with credit points for output measurement puts average efficiency at 70%. Whereas average efficiency in model 1 hardly changes and in model 2 increases modestly in the period 1994-1996, a closer look at single hospitals displays a variety of different efficiency developments over time. PMID- 11860082 TI - Subspecies characterization of porcine Campylobacter coli and Campylobacter jejuni by multilocus enzyme electrophoresis typing. AB - Enteropathogenic Campylobacterjejuni, C. coli and C. lari are currently the most common causes of acute infectious diarrhoeal illness in the UK. Many domestic animals, including pigs, act as natural reservoirs of these organisms and infection may occur through the ingestion of contaminated foodstuffs. C jejuni and C. coli, isolated from the livers of bacon pigs, were examined at subspecies level by multilocus enzyme electrophoresis (MEE) typing with seven enzymic loci. Polymorphological variation was highest with indophenol oxidase, isocitrate dehydrogenase and L-phenylalanyl-L-leucine peptidase giving 5. 5 and 4 alleles at these loci, respectively. The 35 Campylobacter isolates examined in this study (12 C. jejuni and 23 C coli) represented 30 unique electrophoretic types (ETs). Of these ETs, 8 unique types were detected for the 12 C jejuni isolates and 19 unique ETs were detected for the 23 C coli isolates. In addition, 3 types (ETs 2, 5, 10) were shared in common among C. jejuni and C coli. The average number of alleles per enzyme locus was 3.28. The mean genetic diversity, i.e. arithmetic average over all loci assayed, including monomorphic values, was 0.5573 and 0.5350 for C jejuni and C coli. respectively. Alleles were shared by C jejuni and C coli, suggesting an exchange of genetic material between the species. MEE analyses of isolates showed that there was a wide range of subspecies types within both C. jejuni and C coli in porcine livers. In certain cases, up to four phenotypically different strains of C coli were isolated from one liver, indicating multiple infections. PMID- 11860083 TI - Comparative in vitro activity of 16 antimicrobial agents against Actinobacillus pleuropneumoniae. AB - Sixteen antimicrobial agents were tested for their activity against 68 isolates of Actinobacillus pleuropneumoniae by determining the minimum inhibitory concentrations (MICs). Ceftiofur and the fluoroquinolones danofloxacin and enrofloxacin were the most active compounds, with a MIC for 90% of the isolates (MIC90) of (0.05 microg/ml. The MIC90 values of benzylpenicillin, amoxicillin and aspoxicillin were 0.78 units/ml, 0.39 microg/ml and < or = 0.05 microg/ml, respectively. Three isolates (4.4%) were resistant to penicillins, but aspoxicillin was as active as ceftiofur against the susceptible isolates, with MICs of < or = 0.05 microg/ml for all isolates. Resistance to oxytetracycline, chloramphenicol and thiamphenicol occurred in 22 (32.4%), 14 (20.6%) and 15 (22.1%) of the isolates, respectively. Doxycycline was more active than oxytetracycline, with a MIC90 of 1.56 microg/ml as against 25 microg/ml. Florfenicol was not only as active as thiamphenicol, with a MIC for 50% of the isolates (MIC50) of 0.39 microg/ml, but also active against thiamphenicol resistant isolates. All the isolates were susceptible to florfenicol. All the isolates were also susceptible to gentamicin, spectinomycin, tilmicosin, colistin and tiamulin. Of these, spectinomycin was the least active, with a MIC50 of 25 microg/ml, followed by tiamulin, with a MIC50 of 6.25 microg/ml. Of the 68 isolates tested, 49 (72.0%) were of serotype 2; 14 (20.5%) were of serotype 1; 2 each (3.0%) were of serotypes 5 and 6; and one was of serotype 7. Of the isolates, 23 (33.8%) were resistant to one or more of the major antibiotics. Antibiotic resistance was found only infrequently among serotype 2, with 5 (10.2%) of 49 isolates being resistant to chloramphenicol and/or oxytetracycline, while it occurred in 18 (94.7%) of the 19 isolates of other serotypes. PMID- 11860084 TI - Evaluation of the Abbott LCx Mycobacterium tuberculosis assay for direct detection of Mycobacterium bovis in bovine tissue samples. AB - The commercial LCx amplification assay, usually employed to detect the Mycobacterium tuberculosis complex in respiratory specimens, was evaluated by comparing the results it gave with those obtained using Lowenstein-Jensen solid medium and pathological findings in 55 lymph nodes from cattle with positive and 10 lymph nodes from cattle with negative skin tests for tuberculosis. Fifty-three cultures (51 and 2, respectively) were positive for M. bovis, while the results for the LCx assay and the histological method were positive in 48 (45, 3) and 24 (20, 4) samples, respectively. None of the samples from cattle from certified tuberculosis-free herds were positive by any of the procedures. The results obtained with the LCx assay, compared with the culture procedure, regarded as the gold standard among the diagnostic techniques, gave a specificity of 91.6% and sensitivity of 90.5%. Although the sensitivity of LCx was suboptimal, DNA of M. bovis was detected in 81.8% of the skin test-positive animals. Amplification techniques could provide a rapid and reasonably reliable tool for detecting bovine tuberculosis. PMID- 11860085 TI - Chiral inversion of (R)-ketoprofen: influence of age and differing physiological status in dairy cattle. AB - The chiral inversion of ketoprofen has been previously demonstrated in cattle, but no studies have been performed on different ages and metabolic situations in the animals. The aim of this work was to study any modifications of the stereoconversion of ketoprofen that occur by reason of age, lactation or gestation in dairy cows. Holando Argentino cattle were divided into three groups: 8 cows in early lactation, 8 pregnant cows and 8 newborn calves. Four animals from each group received the enantiomer R-(-)-ketoprofen by intravenous administration; the other four animals received the S-(+) enantiomer, all at doses of 0.5 mg/kg. Blood samples were collected at standardized times after dosing and assayed for ketoprofen by high-performance reversed-phase liquid chromatography (HPLC). The percentage inversion of R-(-)-ketoprofen to S-(+) ketoprofen was 50.5% (SD +/- 2.4) in the preruminants, 33.3% (SD +/- 1.7) in cows in early lactation and 26.0% (SD +/- 5.1) in cows in gestation. These results indicate a differing enantioselective metabolic behaviour for one compound in one species under different physiological situations. PMID- 11860086 TI - An experimental study on the treatment and prevention of shimao zheng (fleece eating) in sheep and goats in the Haizi area of Akesai county in China. AB - A series of experimental trials on methods of treatment and prevention of shimao zheng (fleece-eating) were carried out using various sulphates. The results confirmed the hypothesis that this is a local nutritional and metabolic disease, caused mainly by a sulphur deficiency, that could be prevented or cured with supplementary sulphur in the diet. In field studies, the clinical signs virtually disappeared within 14 days of the animals receiving medicated pellets. Over the same period, the concentrations of sulphur in the blood, liver and spleen returned to normal and the pathological changes in the skin improved. The condition of the controls became worse in each of these respects over this time. However, the pathological changes in the musculature were not allevated by the treatment. PMID- 11860087 TI - Some comparative aspects of the pharmacokinetics of tylosin in buffaloes and cattle. AB - The pharmacokinetics of tylosin were compared in cattle (Bos taurus) and buffaloes (Bubalus bubalis). Six animals received each a single dose of 10 mg/kg of tylosin tartrate by the intramuscular route. The serum concentration (Cmax) and the volume of distribution (Vd) presented significant differences between the two species. Cmax was 0.40 +/- 0.046 microg/ml for buffaloes and 0.64 +/- 0.068 microg/ml for cattle. Vd was 1.91 +/- 0.12 L/kg and 1.33 +/- 0.09 L/kg for buffaloes and cattle, respectively. However, as the present study did not show considerable differences in the pharmacokinetics of tylosin in buffaloes and cattle, similar dosage regimes of this drug can be recommended for both species. PMID- 11860089 TI - ISPO consensus conference on appropriate orthopaedic technology for low-income countries: conclusions and recommendations. International Society for Prosthetics and Orthotics. PMID- 11860090 TI - ISPO consensus conference on poliomyelitis: consensus statements. International Society for Prosthetics and Orthotics. PMID- 11860088 TI - Pharmacokinetics of oleandomycin in dogs after intravenous or oral administration alone and after pretreatment with metamizole or dexamethasone. AB - The pharmacokinetics of oleandomycin (OLD) after intravenous and oral administration, both alone and after intramuscular pretreatment with metamizole or dexamethasone, were studied in healthy dogs. After intravenous injection of OLD alone (10 mg/kg as bolus), the elimination half-life (t 1/2 beta, volume of distribution (Vd,area), body clearance (ClB) and area under the concentration time curve (AUC) were 1.60 h, 1.11 L/kg. 7.36 (ml/kg)/min and 21.66 microg h/ml, respectively. There were no statistically significant differences following pretreatment with metamizole or dexamethasone. After oral administration of OLD alone, the t 1/2 beta, maximum plasma concentrations (Cmax), time of Cmax (tmax), mean absorption time (MAT) and absolute bioavailability (Fabs) were 1.6 h, 5.34 microg/ml, 1.5 h, 1.34 h and 84.29%, respectively. Pretreatment with metamizole caused a significantly decreased value for Cmax (2.93 microg/ml) but the MAT value (2.23 h) was significantly increased. Statistically significant changes in the pharmacokinetic parameters of OLD following oral administration were also observed as a result of pretreatment with dexamethasone. The Cmax was increased (8.24 microg/ml) and the tmax (0.5 h) and MAT (0.45 h) were lower. PMID- 11860091 TI - Lower limb amputations: registration of all lower limb amputations performed at the University Hospital of Trondheim, Norway, 1994-1997. AB - The authors consecutively recorded all lower limb amputations performed at the University Hospital of Trondheim from January 1st, 1994 to January 1st, 1997. A total of 215 primary lower limb amputations were carried out in the study period: 40 partial foot amputations, 2 ankle disarticulations, 51 trans-tibial amputations, 68 knee disarticulations, 50 transfemoral amputations and 4 hip disarticulations. Seventy-four (74) (34%) of the amputees had diabetes mellitus, 113 (53%) had peripheral vascular disease and 28 (13%) of the amputees had various diagnoses. In those who were amputated due to diabetic and peripheral vascular disease the overall reamputation rate was 19% and 20%, respectively. The rates of reamputation at the trans-tibial and knee level were similar. In the city of Trondheim the annual incidence of primary amputations was 34 per 100,000 and 4.4 per 1,000 diabetic subjects. The incidence of lower limb amputations was 25 times higher in diabetic subjects compared to non-diabetic subjects. PMID- 11860092 TI - Consequences of non-vascular trans-femoral amputation: a survey of quality of life, prosthetic use and problems. AB - Individuals with unilateral trans-femoral amputations due to non-vascular causes were studied in a mailed survey designed to investigate health-related quality of life (HRQL), prosthetic use and problems. The Swedish SF-36 Health Survey and a structured questionnaire designed for trans-femoral amputees were used. The series consisted of 97 subjects (60 men, 37 women), aged 20 to 69 years with a mean of 22 years since the amputation. Trauma was the cause of amputation in 55%, tumour in 35% and other causes in 10%. Ninety-two (92) subjects (95%) had a prosthesis and 80 (82%) used it daily. General HRQL was significantly lower than Swedish age- and gender-matched norms in all dimensions as measured by SF-36. Most frequently reported problems that had led to reduction in quality of life were heat/sweating in the prosthetic socket (72%), sores/skin irritation from the socket (62%), inability to walk in woods and fields (61%) and inability to walk quickly (59%). Close to half were troubled by stump pain (51%), phantom limb pain (48%), back pain (47%) and pain in the other leg (46%). One fourth considered themselves to have a poor or extremely poor overall situation. Transfemoral amputation, due to non-vascular causes, has an evident impact on quality of life and there are considerable problems related to the amputation and the prosthesis. Efforts to improve the physical and the psychological well-being for this group, with a long life expectancy, are needed. PMID- 11860093 TI - Effect of trans-tibial prosthesis pylon flexibility on ground reaction forces during gait. AB - This study explored the effects of trans-tibial prosthesis pylon flexibility on ground reaction forces (GRFs) associated with walking and step-down. Four (4) active subjects with unilateral trans-tibial amputation and pylon lengths ranging from 4.9 cm to 25.9 cm were studied wearing an aluminium (rigid) pylon and a nylon (more flexible) pylon. Ground reaction forces were collected for the amputated limb during walking at pre-measured self-selected velocity and when stepping down from a 20 cm box. Pylon material significantly affected the magnitudes and patterns of GRFs in both tests. During walking, the most notable differences were seen in the anteroposterior (AP) direction. With the flexible pylon, the AP propulsive peak was greater (p=0.031), and the irregularities in the AP force curve were reduced. Additionally, when walking with the flexible pylon, the vertical peak associated with weight acceptance occurred earlier (p=0.010), the vertical terminal stance peak occurred later (p=0.012), and stance time was longer (p=0.010). During step-down, the vertical loading rate (p=0.010) and the peak vertical force (p=0.010) were greater with the more flexible pylon. Subjective feedback indicated that subjects could distinguish between the two pylons and felt that the nylon component was more comfortable, more flexible, and would enable them to walk more quickly. These results suggest that the pylon may be an influential component of the prosthesis with respect to gait and comfort, and that some degree of flexibility is desirable. PMID- 11860094 TI - A comparison of trans-tibial amputee suction and vacuum socket conditions. AB - Daily volume loss of the stump leads to a poor fit of the prosthetic socket. A method of preventing this volume loss and maintaining a good fit was developed. A vacuum (-78 kPa) was drawn on the expulsion port of a total surface-bearing suction socket to hold the liner tightly against the socket. Stump volume of 10 trans-tibial amputees was measured prior to and immediately after a 30 minute walk with normal and vacuum socket conditions. Under the normal condition, the limb lost an average of 6.5% of its volume during the walk. In contrast, with the liner held tightly by vacuum, the limb gained an average of 3.7% in volume. It is believed that the difference observed between conditions resulted from a greater negative pressure developed during the swing phase of gait with the vacuum condition. X-rays revealed that the limb and tibia pistoned 4 mm and 7 mm less, respectively, under the vacuum condition. The combination of reduced pistoning and maintenance of volume is thought to account for the more symmetrical gait observed with the vacuum. PMID- 11860095 TI - Optimising the trans-femoral prosthetic alignment for running, by lowering the knee joint. AB - For amputees to perform an everyday task, or to participate in physical exercise, it is crucial that they have an appropriately designed functional prosthesis. The aim of this study was to investigate the optimal trans-femoral prosthetic alignment configuration for running. A case study design was implemented as the method to collect data on four male Paralympic level trans-femoral amputee runners. In total 28 synchronised 3D kinematic, kinetic, and temporal biomechanical measures were analysed. A new prosthetic alignment, in which the prosthetic knee axis was lowered longitudinally (moved distally), was evaluated for running. The performance of the symmetry indices and running time for a total of three new modified alignments were compared to the standard prosthetic alignment. The interlimb asymmetry was found to improve when the subjects ran on the modified prosthetic alignment, and most importantly a one way ANOVA found a statistically significant increase in running velocity. This study identified that for all four subjects, who used the same prosthetic components, lowering the prosthetic knee joint centre improved their interlimb symmetry, and subsequently their running velocity by an average of 26%. PMID- 11860096 TI - Ultrasound study of the motion of the residual femur within a trans-femoral socket during daily living activities other than gait. AB - This study analyses the residual femur motion of a single amputee within a transfemoral socket during a series of daily living activities. Two simultaneously transmitting, socket mounted transducers were connected to two ultrasound scanners. Displacement measurements of the ultrasound image of the femur were video recorded and measured on "paused" playback. Abduction/adduction and flexion/extension of the residual femur within the socket at any instant during these activities were estimated, knowing the relative positions of the two transducers and the position of the residual femur on the ultrasound image. Consistent motion patterns of the residual femur within the trans-femoral socket were noted throughout each monitored daily living activity of the single amputee studied. Convery and Murray (2000) reported that during level walking, relative to the socket, the residual femur extends 6 degrees and abducts 9 degrees by mid stance while flexing 6 degrees and adducting 2 degrees by toe-off. Uphill/downhill, turning to the right and stepping up/down altered this reported pattern of femoral motion by approximately 1 degree. During the standing activity from a seated position the femur initially flexed 4 degrees before moving to 7 degrees extension, while simultaneously adducting 6 degrees. During the sitting activity from a standing position the femur moved from 7 degrees extension and 6 degrees adduction to 3 degrees flexion and 1 degree abduction. The activity of single prosthetic support to double support introduced only minor femoral motion whereas during the activity of prosthetic suspension the femur flexed 8 degrees while simultaneously adducting 9 degrees. Additional studies of more amputees are required to validate the motion patterns presented in this investigation. PMID- 11860097 TI - Prosthetic management of children in The Netherlands with upper limb deficiencies. AB - The aim of the study was to assess which children with congenital and acquired upper limb deficiencies were fitted with prostheses, what types of prostheses were prescribed as first, second and third prostheses, at what age prostheses were first prescribed and how long the children wore their prostheses. The design was a retrospective chart review at De Hoogstraat Rehabilitation Centre, Utrecht (The Netherlands). Medical files of all patients with congenital or acquired upper limb deficiencies who visited the outpatient clinic between 1972 and 1996 were reviewed, collecting data on patient characteristics, prosthesis prescription and use of prostheses. The group included 224 children, of whom 206 (92%) had congenital deficiencies. Of all children with unilateral congenital deficiencies, 54% had been fitted with prostheses, against 3% of all children with bilateral congenital deficiencies and 67% of all children with acquired deficiencies. In the congenital group, it was children with transverse defects of one-third or two-thirds of the forearm who had most frequently had prostheses fitted (85% of the children). Most of the children with unilateral congenital deficiencies had received passive prostheses as their first prostheses (80 of the 90 prescribed prostheses); children with acquired defects usually had active prostheses (8 of the 12 prescribed prostheses). Body-powered prostheses were most commonly prescribed as the second type of prosthesis. In the group of 119 children who had been seen before the age of 4 years and had been followed for at least three years, 63 had been fitted with one or more prostheses at a mean age of 2.6 (SD 2.5) years. Of the 46 children with congenital defects, 30 had been fitted with prostheses, and at the age of 12, two-thirds of them still used their prostheses (63%, 19/30). "De Hoogstraat" rehabilitation centre uses a restrained prosthesis prescription policy, depending on the type of deficiency and the expected functional benefits. Data on prosthesis use are encouraging, although a follow-up study is required to determine the functional outcome for prosthesis users and non-users. PMID- 11860098 TI - Characterising phantom limb phenomena in upper limb amputees. AB - A systematic descriptive survey was carried out on a clinic population with acquired upper limb amputation in which clear distinctions were made between phantom experiences, pre-amputation pain and stump pain. It was found that of 76 participants 96% reported phantom experiences and 84% were currently experiencing phantom limb phenomena (PLP). Sixty-nine percent (69%) of those currently experiencing phantom phenomena stated that the phantoms were painful. Significant relationships were found between frequency of phantom experiences and side and level of amputation. In addition, the presence of 'trigger' points reported by a sub group of participants was found to be associated with the frequency of phantoms and painful phantoms. This finding could provide support for cortical reorganisation post amputation. It is suggested that detailed individual 'profiling' of phantom experiences is important and would have the following implications: it would contribute to existing knowledge and provide new insights into the central nervous system's reorganisation post-amputation; it would provide an accurate way of representing amputees' anomalous perceptions; it would alert health professionals to the presence, variety and extent of PLP following amputation. PMID- 11860099 TI - An unusual complication of a myoelectric prosthesis. AB - This clinical note describes an unusual and previously unreported complication of wearing a myoelectric prosthesis in a child with congenital upper limb deficiency. After an initial period of wearing a cosmetic prosthesis, he was provided with a child's trans-radial myoelectric prosthesis with a Steeper Scamp Electric hand at the age of 21 months. After successful and uncomplicated use of this prosthesis for 2 years--a sudden onset of burns of the skin of the stump underlying the single electrode site was noted. The cause of the burns was thought to be due to heat generated from electrical failure possibly from ingress of moisture. This complication has not before been experienced in the authors' centre, nor has it been reported in the literature. While constant improvements are being made in the field of electrically powered prosthesis, the rehabilitation team should be aware of this unusual complication. PMID- 11860101 TI - Effect of transglutaminase on the heat stability of milk: a possible mechanism. AB - Treatment of milk with transglutaminase (TGase) affects its heat stability, but the manner in which it does so depends on whether or not the milk had been preheated before incubation and on the temperature of preheating. In raw milk, it appears that cross-link formation between the individual caseins is responsible for preventing the dissociation of kappa-casein from the micelles at pH values in the region of minimum stability. In milks preheated before incubation with TGase, denaturation of whey protein may have allowed the formation of cross-links by TGase between denatured whey proteins and the individual caseins which, in combination with cross-linking of the caseins, contributed to greatly improved heat stability at pH > 6.5. It appears from the results of this study that TGase has potential commercial applications as a food-grade additive capable of improving the heat stability of milk. PMID- 11860102 TI - Reproduction, mastitis, and body condition of seasonally calved Holstein and Jersey cows in confinement or pasture systems. AB - Dairy cows in confinement and pasture-based feeding systems were compared across four spring-calving and three fall-calving replicates for differences in reproduction, mastitis, body weights, and body condition scores. Feeding systems and replicates included both Jersey and Holstein cows. Cows in confinement were fed a total mixed ration, and cows on pasture were supplemented with concentrates and provided baled hay or haylage when pasture supply was limiting. Breeding periods were for 75 d in spring or fall. Reproductive performance did not differ significantly due to feeding system or season. Jerseys had higher conception rates (59.6 vs. 49.5 +/- 3.3%) and higher percentages of cows pregnant in 75 d (78.1 vs. 57.9 +/- 3.9%) than Holsteins. Cows in confinement had 1.8 times more clinical mastitis and eight times the rate of culling for mastitis than did cows on pasture. Jerseys had half as many clinical cases of mastitis per cow as Holsteins. Only 41 +/- 5% of confinement Holsteins remained for a subsequent lactation, starting within the defined calving season compared with 51 +/- 5% of pastured Holsteins and 71 and 72 +/- 5% of Jerseys, respectively. Body weights and condition scores were generally higher for confinement cows than pastured cows, and Jerseys had higher condition scores and lower body weights than Holsteins. In summary, pastured cows had fewer clinical cases of mastitis, lower body condition scores, and lower body weights than confinement cows. Holsteins were less likely to rebreed, had more mastitis, higher culling rates, and lower body condition scores than Jerseys. PMID- 11860100 TI - A series of case studies on the effect of a midfoot control ankle foot orthosis in the prevention of unresolved pressure areas in children with cerebral palsy. AB - This paper reports on a series of case studies where improvements were sought in muscle tone and gait in children with cerebral palsy. A Midfoot Control Ankle Foot Orthosis (AFO) was developed to control foot position in a cohort of patients with cerebral palsy (CP). The concept of controlling midfoot and hindfoot with an encapsulated internal Supra-Malleolar AFO that fitted into an external AFO was shown to be effective in ambulant children with CP. Some initial problems of compliance were noted and postulated to be due to difficulties associated with previous orthotic devices. Evidence from the case studies suggest that the developed Supra-Malleolar AFO orthoses enables children with CP to maintain mobility without skin tissue damage, delays the need for surgery and at the same time maintains the length of the Triceps Surae (Gastrocnemius and Soleus) complex. Plans for further research are discussed which will contribute to the evidence base for this particular orthotic device. PMID- 11860103 TI - The effect of selective dry cow treatment on new intramammary infections. AB - Dry cow therapy, or antibiotic treatment at end of lactation, is used to eliminate intramammary infections and prevent new infections during the dry period. It is one part of a total management system recommended in controlling intramammary infections in the dairy cow. Public health concerns advise prudent use of antibiotics, as their use may promote bacterial antibiotic resistance and leave antibiotic residues in the food chain. The effects of dry cow treatment and no treatment were compared, on new intramammary infections and clinical mastitis within two low cell count herds and two herds undergoing conversion to organic farming. The results will inform those restricting their use of dry cow therapy on the additional risk of new intramammary infection and aid in development of alternative management strategies. No cases of clinical mastitis in the dry period were observed in treated cows, whereas in the untreated groups a significant number were observed. Significantly more new infections at calving were found in the untreated group in all herds. In those quarters where infections were first detected at calving, the incidence of clinical mastitis was significantly greater in the untreated group in all herds. Clinical mastitis detection was significantly lower in organic herds. Untreated quarters infected at drying with Corynebacterium spp. or coagulase-negative staphylococci were found to have an increased risk of new infection by Streptococcus uberis or coliform bacteria. It can be concluded that dry cow therapy continues to lower significantly the rate of new dry period intramammary infection in herds with elevated somatic cell counts and a high prevalence of infection. PMID- 11860105 TI - Antibiotic susceptibility patterns for environmental streptococci isolated from bovine mastitis in central California dairies. AB - Environmental streptococci are frequently isolated from bovine mastitis in dairy cows with only limited information available on the antimicrobial susceptibility of these organisms. A total of 362 environmental streptococci isolated from cases of bovine mastitis from the central San Joaquin Valley of California over a 3-yr period were used in the study. Overall, 39.9% of the strains tested were Streptococcus uberis, 42.2% were Streptococcus dysgalactiae, and 11.1% were Enterococcus spp. The antimicrobial susceptibility for these organisms was determined for the following antimicrobial agents: penicillin, ampicillin, cephalothin, ceftiofur, penicillin + novobiocin, erythromycin, pirlimycin, tetracycline, and sulfadimethoxine. Results demonstrate substantial differences in the susceptibility patterns for the various organisms collectively referred to as the environmental streptococci. The MIC90 for penicillin was 0.06 microg/ml for 152 strains of S. dysgalactiae compared with 0.25 microg/ml for 133 strains of S. uberis. However, the Enterococcus spp. were the most resistant organisms tested. These data also indicate that the use of interpretive criteria based on human data may provide misleading results. In conclusion, these data confirm that the environmental streptococci are a diverse group of organisms comprised of several different genera and species and that identification of environmental streptococci to the species level is needed to appropriately modify control methods. Moreover, the use of the agar disk diffusion (Kirby-Bauer) susceptibility test for agents with human-based interpretive criteria is contraindicated, and these tests should only be performed with agents with mastitis specific interpretive criteria. PMID- 11860104 TI - Use of estradiol cypionate in a presynchronized timed artificial insemination program for lactating dairy cattle. AB - Experiment 1 evaluated pregnancy rates when estradiol cypionate (ECP) was used to induce ovulation as part of a timed artificial insemination (TAI) protocol in comparison to Ovsynch for lactating dairy cows in Florida (n = 371) and Texas (n = 321). Cows were presynchronized with two injections of PGF2, (25 mg, im) given 14 d apart with TAI protocols beginning 14 d after the second injection of PGF20. The TAI protocols consisted of an injection of GnRH (100 microg, im) followed by PGF2alpha 7 d later. Then, cows either received an injection of GnRH (Treatment I, Ovsynch) at 48 h after PGF2alpha and inseminated 16 to 24 h later or received an injection of ECP (1 mg, i.m.) at 24 h after PGF2alpha, (Treatment II; Heatsynch) and inseminated 48 h later. In Florida, pregnancy rates after TAI were 37.1 +/- 5.8% for Ovsynch compared with 35.1 +/- 5.0% for Heatsynch. In Texas, pregnancy rates were 28.2 +/- 3.6% for Ovsynch and 29.0 +/- 3.5% for Heatsynch. Overall pregnancy rates did not differ between Ovsynch and Heatsynch treatments. In Experiment 2, estrus and ovulation times were determined in lactating dairy cows submitted to the Heatsynch protocol. Frequencies of detected estrus and ovulation after ECP were 75.7% (28/37) and 86.5% (32/37), respectively. Mean intervals to ovulation were 55.4 +/- 2.7 h (n = 32) after ECP and 27.5 +/- 1.1 h (n = 27) after onset of estrus. Estrus occurred at 29.0 +/- 1.8 h (n = 28) after ECP. It is recommended that any cow detected in estrus by 24 h after ECP injection be inseminated at 24 h and all remaining cows be inseminated at 48 h because 75% (n = 24/32) of the ovulations occurred between > or = 48 h to < or = 72 h after ECP. Synchronization of ovulation and subsequent fertility indicated that estradiol cypionate could be used to induce ovulation for successful timed insemination. PMID- 11860106 TI - Apoptosis of bovine neutrophils following diapedesis through a monolayer of endothelial and mammary epithelial cells. AB - In a two-chamber system, isolated blood polymorphonuclear neutrophil leukocytes (PMN) were allowed to migrate (5 h, 37degrees C) in response to bovine complement component C5a across calfskin and rat-tail type I collagen-coated micropore membranes, arterial endothelial, or mammary epithelial cell monolayer on calfskin and rat-tail collagen-coated membranes, respectively. Migration through calfskin collagen-coated membranes resulted in 14.5% +/- 3.4% apoptotic PMN, which was significantly higher than 6.6% +/- 1.2% apoptotic nonmigrated C5a-treated PMN. The addition of an endothelial or epithelial cell monolayer to collagen-coated membranes prevented apoptosis of migrated PMN. After removing the membranes, nonmigrated (untreated and C5a treated) and migrated PMN were incubated for an additional 20 h. At this time point, 69.1% +/- 4.5% and 47% +/- 4.5% of PMN that have migrated through a calfskin-coated membrane and an endothelial monolayer, respectively, were apoptotic, compared with 28.2% +/- 3.0% and 21.1% +/- 4.5% apoptotic untreated and C5a-treated PMN, respectively; 46.9% +/- 4.8% of PMN that have migrated through rat-tail-coated membranes were apoptotic compared with 14.7% +/- 2.3% and 9.3% +/- 1.2% apoptotic untreated and C5a-treated PMN, respectively. Migration across rat-tail collagen-coated membranes with a monolayer of epithelial cells did not affect apoptosis of migrated PMN, even after 20 h of incubation. In conclusion, migration of PMN across collagen-coated membranes (either calfskin or rat-tail collagen) induced an apoptotic response, which was downregulated by a monolayer of endothelial cells and was negated by an epithelial cell monolayer. PMID- 11860107 TI - Effect of carbon dioxide on the growth of Bacillus cereus spores in milk during storage. AB - The effects of the addition of 11.9 mM CO2 on the growth of Bacillus cereus spores inoculated into sterile homogenized whole milk at 101 and 106 spores/ml and stored at 6.1 degrees C, was examined weekly for 35 d. Colony-forming units from CO2 treated inoculated milks decreased over 35 d at a rate similar to that of untreated inoculated milk, as defined by linear regression. Plate counts for treated and control milks inoculated at 10(1) cfu/ml were not significantly different on sampling d 0, 14, 21, and 28. Plate counts at d 7 were significantly different and counts at d 35 were at undetectable levels for both treated and control milks. Plate counts for milk inoculated at 10(6) cfu/ml were not significantly different on d 0, 28, and 35; they were significantly different on d 7, 14, and 21. There was no consistency as to whether the control or test milks were higher in counts on days when the differences were significant. Added CO2 reduced the pH of the milk from an average value of 6.61 to an average value of 6.31; however, this drop did not correlate with changes in any other parameter measured. These data suggest that moderate levels of CO2 do not enhance the outgrowth of B. cereus spores over long-term storage and do not increase the risk of foodborne illness due to the organism. PMID- 11860108 TI - Effects of level of concentrate feeding during the second gestation of Holstein Friesian dairy cows. 1. Feed intake and milk production. AB - The objective of this experiment was to examine the residual effects of the level of concentrate feeding during the second gestation on performance in the second lactation. Forty-three Holstein-Friesian heifers that calved for the first time at 2 or 3 yr of age were offered diets based on ad libitum consumption of ryegrass silage and either 2 or 7 kg/d of concentrates over the second half of the first lactation. All cows received a low quality diet, based on grass silage and barley straw (60:40 DM basis) for a short (6-wk) dry period. Feeding in the second lactation was based on ad libitum access to grass silage and a flat rate of concentrates (8 kg/ d for 120 d; 5 kg/d thereafter). Three-year-old heifers produced more milk than 2-yr old heifers in the first lactation, equivalent to 93 kg of fat-corrected milk for each additional month of rearing. Cows given the low level of concentrates produced less milk and gained less weight and body condition, though there was a partial compensation when silage quality improved in late lactation. Forage intake declined as cows approached calving, and cows gained little weight or body condition over the dry period. There was no effect on calf weights. Cows that had received the low level of concentrates in the previous lactation consumed more forage dry matter during the second lactation. Parallel curve analysis showed that 2-yr-old heifers that had received a low level of concentrates produced significantly less milk in the second lactation. The numerically much greater gains of weight and body condition for this group were not statistically significant, owing to large between-cow variation. PMID- 11860109 TI - Effects of level of concentrate feeding during the second gestation of Holstein Friesian dairy cows. 2. Nitrogen balance and plasma metabolites. AB - We offered 43 Holstein-Freisian dairy heifers that calved for the first time at either 2 or 3 yr of age ad libitum access to ryegrass silage and a standard concentrate allowance of either 2 or 7 kg/d from the middle to the end of their first lactation. All animals were given the same relatively poor quality dry period diet of a mixture of ryegrass silage and barley straw (63:37 dry matter basis) from 6 wk before their predicted second-calving date. Following their second calving, all animals received access to the same ration of ad libitum grass silage and concentrates at a rate of 8 kg/d to 120 d of lactation and 5 kg/d thereafter until the end of the experimental recording at about 150 d of lactation. Nitrogen balance was significantly higher at the end of the first lactation for animals that were given the higher concentrate allowance and tended to be higher for older animals. There was no effect of age or residual effect of concentrate allowance on N balance during the dry period or during the second lactation. Labile body protein reserves, as estimated by the depth of the muscle Longissimus dorsi (which was significantly correlated with body condition score), were similar for all animals during the dry period, but younger animals previously offered the lower concentrate allowance did not lose L. dorsi depth early in the second lactation as did other animals. Arterial plasma concentrations of amino acids Phe, Trp, and Leu were significantly higher in younger animals at wk 8 of the second lactation, and Gly was significantly lower, although mammary blood flow, arteriovenous differences, and rates of uptake of the AA measured were unaffected by treatment. It is concluded that differences in second-lactation milk yield were not mediated through the availability of labile body protein or the supply of nutrients to the mammary gland. PMID- 11860110 TI - Effects of high intensity pulsed electric field and thermal treatments on a lipase from Pseudomonas fluorescens. AB - Milk and dairy products may contain microorganisms capable of secreting lipases that cause sensory defects and technological problems in the dairy industry. In this study, the effects of thermal and high-intensity pulsed electric field (HIPEF) treatments on an extracellular lipase from Pseudomonas fluorescens, suspended in a simulated skim milk ultrafiltrate (SMUF) have been evaluated. Heat treatments applied were up to 30 min from 50 to 90 degrees C. HIPEF treatments were carried out using pilot plant facilities in a batch or continuous flow mode, where treatment chambers consisted of parallel and coaxial configuration, respectively. Samples were subjected to up to 80 pulses at electric field intensities ranging from 16.4 to 37.3 kV/cm. This resulted in a lipase that was quite resistant to heat and also to HIPEF. High (75 degrees C-15 s) and low pasteurization treatments (63 degrees C-30 min) led to inactivations of 5 and 20%, respectively. Using the batch-mode HIPEF equipment, a 62.1% maximum activity depletion was achieved after 80 pulses at 27.4 kV/cm. However, when HIPEF treatments were applied in the continuous flow mode, an inactivation rate of just 13% was achieved, after applying 80 pulses at 37.3 kV/cm and 3.5 Hz. The results of both heat and HIPEF treatments on enzyme inactivation were adjusted with good agreement to a first-order kinetic model (R2 > 62.3%). PMID- 11860111 TI - Occurrence of trans-C18:1 fatty acid isomers in goat milk: effect of two dietary regimens. AB - Trans-octadecenoic acid composition of goat milk fat was studied by using silver ion thin layer chromatography combined with gas-liquid chromatography. This analytical procedure also was used to investigate the effect of diet on trans C18:1 fatty acid content in goat milk. Thirty-two goats were used in a 2 x 2 factorial arrangement and treatments. Two groups of goats received alfalfa hay at either a high or a low level of forage and two other groups received Rumiluz (dehydrated alfalfa from France Luzerne, Chalon en Champagne, France) at either a high or a low level of forage. Trans-C18:1 isomer proportions (relative to total fatty acids) were, respectively, 2.02% for the Rumiluz low-level group and 1.75% for the Rumiluz high-level group versus 1.71% for the alfalfa low-level group and 1.21% for the alfalfa high-level group. Goats fed on Rumiluz thus produced relatively higher levels of trans-C18:1 fatty acids than animals fed alfalfa hay. The results also showed that production of trans-C18:1 fatty acids increased when the level of forage in the diet decreased. Moreover, goat milk trans-C18:1 composition appeared similar to the cow milk profile. Vaccenic acid, trans-11 C18:1, was the major component and represented about 36.2% of total trans-C18:1 isomers. PMID- 11860112 TI - Effects of spray-dried whole egg and biotin in calf milk replacer. AB - Holstein bull calves (n = 120) were fed milk replacers containing 0, 10, or 20% of the formulation (0, 22, or 44% of crude protein) as spray-dried whole egg powder in a 56-d feeding trial. Milk replacer was medicated with oxytetracycline and neomycin and was fed from d 1 to 42 of the study in a phase-fed program. All experimental milk replacers were supplemented with B vitamins, except biotin. One half of all calves were supplemented with 1 mg/kg of supplemental biotin to determine whether avidin in the egg protein product inhibited growth. Increasing spray-dried whole egg caused a linear reduction in body weight, body weight gain at 28 and 56 d of the study, calf starter intake, and feed efficiency. Calves fed milk replacers containing 0, 10, and 20% spray-dried whole egg gained an average of 486, 369, and 302 g/d, respectively, during the 56-d trial. Efficiency of feed utilization was 446, 318, and 231 g of body weight gain per kilogram of dry matter intake. Improvement in body weight and feed efficiency occurred when calves began consuming calf starter on d 29. Digestibility of protein or fat from egg may have been reduced during the trial; however, the addition of biotin to the milk replacer did not influence animal performance, suggesting that avidin in spray-dried whole egg was not responsible for impaired performance. The spray dried whole egg product used in this study did not provide nutrients to support adequate growth of milk-fed calves. PMID- 11860113 TI - Effects of abomasal infusions of histidine, glucose, and leucine on milk production and plasma metabolites of dairy cows fed grass silage diets. AB - Our previous study showed that His was the first-limiting amino acid (AA) for milk protein production in cows fed grass silage and cereal-based supplement. The aim of this study was to identify the second-limiting AA and determine whether glucose was limiting responses to His. Abomasal infusion of His (6.5 g/d), glucose (250 g/d), His (6.5 g/d) + glucose (250 g/d), His 6.5 g/d) + Leu (12 g/d) and His (6.5 g/d) + Leu (12 g/d) + glucose (250 g/d) on milk production and utilization of amino acids by mammary gland was in an incomplete 5 x 6 Latin square design with 14-d periods. The diet was based on restrictively fermented grass silage fed ad libitum and 8 kg/d of concentrate comprised of barley, oats, unmolassed sugar beet pulp, urea, and minerals. The infusions did not affect feed intake, diet digestibility, or rumen fermentation pattern. The molar proportion of propionate in rumen VFA was low (15.5%), suggesting that glucose supply from the basal diet could be limiting. Milk and milk protein yields were increased by His infusion. Infusion of His increased plasma His concentration from 19 to 52 microM but decreased extraction efficiency of His. Infusion of glucose increased plasma glucose concentration, milk lactose concentration, and yield and tended to increase milk protein yield. Responses in milk protein yield to combined infusions of His and glucose were additive, suggesting that the utilization of the first-limiting AA His was limited by glucose supply. Infusion of Leu increased plasma Leu concentration but did not produce any further milk protein yield response compared with the infusions without Leu. It was concluded that the efficiency of utilization of the first-limiting AA His could be improved by increasing the supply of glucose, when the basal diet produces a rumen fermentation pattern low in propionate. Leu was not the second-limiting AA in cows fed grass silage-based diets. PMID- 11860114 TI - Influence of corn processing and frequency of feeding on cow performance. AB - Twenty cows, including five fitted with rumen cannulae, were used to study the influence of corn processing and frequency of feeding on milk yield and ruminal fermentation characteristics. Cows were assigned to five treatments in a 5 x 5 Latin square experiment. Each period was 3 wk. Cows were fed 45% forage and 55% grain in a total mixed ration. Diets contained 35% corn either coarsely ground and fed once a day (1x), finely ground (FGC) fed 1x, steam-flaked (SFC) fed lx, FGC fed four times a day (4x), or SFC fed 4x. Processing of corn and frequency of feeding had no influence on dry matter intake. Digestibility of starch was increased 6 and 3 percentage units by feeding SFC corn compared with coarsely and finely ground corn, respectively. Cows fed SFC or FGC produced 4% more milk with lower fat content compared with coarsely ground corn. Increasing the feeding frequency did not improve milk fat content. The fat-corrected milk yield was not different among treatments. Feeding SFC resulted in a low acetate-to-propionate ratio in the rumen fluid than FGC. Cows fed SFC produced 45 and 115 g more milk protein per cow/d than cows fed FGC or coarse, respectively. With the value of increased milk protein observed in this study, it would be more economical to feed SFC or finely ground corn to dairy cows compared with coarse ground. The breakeven price of flaking corn in this study was $32 and $12/metric tonne compared with coarse and FGC, respectively. Based on a survey conducted by the authors, the price of flaking corn in the United States ranged between $7 to $22/metric tonne during year 2000. PMID- 11860115 TI - Evaluation of models to estimate urinary nitrogen and expected milk urea nitrogen. AB - Milk urea nitrogen (MUN) has been introduced as a means to estimate urinary nitrogen (N) excretion and protein status of dairy cattle. For Holstein cows, the amount of urinary N excreted (g/d) was originally reported to be 12.54 x MUN (mg/dl), but recently urinary N (g/d) was reported to equal 17.64 x MUN (mg/dl). The objectives of the present study were to evaluate models to predict urinary N and expected MUN, by using older and newer data sets, and to quantify changes that may have occurred in MUN measurements over time. Two data sets were used for model evaluation. Data set 1 was from the spring of 1998 and data set 2 was from the spring of 1999. Similar cows and diets were used in both studies. By using data set 1, the newer model underestimated MUN by an average of 3.8 mg/dl, whereas the older model was accurate. By using data set 2, the older model overestimated MUN by 4.8 mg/ dl, but the newer model was accurate. In the period between the two studies, the MUN measured appeared to decrease by an average of 4.0 mg/dl. By using current wet chemistry methods to analyze for MUN, urinary N (mg/dl) can be predicted as 0.026 x MUN (mg/dl) x body weight (kg). Because of changes in methodology that occurred in the fall of 1998, target MUN concentrations have decreased to 8.5 to 11.5 mg/dl for most dairy herds compared with previous target concentrations of 12 to 16 mg/dl. PMID- 11860116 TI - Fish oil and extruded soybeans fed in combination increase conjugated linoleic acids in milk of dairy cows more than when fed separately. AB - Eight multiparous Holstein and four multiparous Brown Swiss (78 +/- 43 DIM) cows were used in a 4 x 4 Latin square with 28-d periods to evaluate if feeding fish oil with a source of linoleic acid (extruded soybeans) would stimulate additional amounts of conjugated linoleic acid in milk. Four treatments consisted of a control diet with a 50:50 ratio of forage to concentrate (DM basis), a control diet with 2% added fat from either menhaden fish oil or extruded soybeans, or a combination of fish oil and extruded soybeans each adding 1% fat. DM intake (24.3, 21.6, 24.5, and 22.5 kg/d, for control, fish oil, extruded soybeans, and combination diets, respectively), milk production (32.1, 29.1,34.6, and 31.1 kg/d), and milk fat content (3.51, 2.79, 3.27, and 3.14%) were lower for cows that consumed either fish oil-containing diet, especially the 2% fish oil diet. The proportion of n-3 fatty acids in milk fat increased similarly among all three fat-supplemented diets. Concentrations of transvaccenic acid (1.00, 4.16, 2.17, and 3.51 g/100 g of fatty acids) and cis-9, trans-11 conjugated linoleic acid (0.60, 2.03, 1.16, and 1.82 g/100 g of fatty acids) in milk fat increased more with fish oil than with extruded soybeans. When fed the combination diet, these fatty acids were approximately 50% higher than expected for Holsteins, whereas concentrations were similar for Brown Swiss compared with feeding each fat source separately. These data indicated that fish oil modifies ruminal or systemic functions, stimulating increased conversion of linoleic acid into transvaccenic and conjugated linoleic acids. PMID- 11860117 TI - Trends in reproductive performance in Southeastern Holstein and Jersey DHI herds. AB - Trends in average days open and services per conception from 1976 to 1999 were examined in 532 Holstein and 29 Jersey herds from 10 Southeastern states. Three year averages for eight intervals (time) were calculated (first: 1976 to 1978; eighth: 1997 to 1999). Milk, fat, fat-corrected milk, and number of cows increased across time. Herds of both breeds had linear, quadratic, and cubic effects of time on days open and services per conception. For 1976 to 1978, respective averages of days open and services per conception were 122 +/- 2.8 d and 1.91 +/- 0.08 for Jerseys, 124 +/- 0.7 d and 1.91 +/- 0.02 for Holsteins. Days open increased nonlinearly to 152 +/- 2.8 d for Jerseys and 168 +/- 0.7 d for Holsteins by 1997 to 1999, resulting in a breed x time interaction. Services per conception also increased nonlinearly, reaching 2.94 +/- 0.04 services for both breeds in 1994 to 1996, changing only slightly after 1996. Fat-corrected milk and number of cows had small but significant effects. Five subregions (one to three states) differed in mean days open and services per conception, but changes in those measures across time among subregions were similar. Days to first service increased by 16 (Holsteins) and 18 d (Jerseys) during the last five 3-yr periods, associated with increasing days open. Estrus detection rates generally declined from 1985 to 1999, associated inversely with services per conception. Reduced reproductive performance in Southeastern dairy herds is of concern. Multiple strategies are needed to attenuate further declines. PMID- 11860118 TI - Efficacy of two acidified chlorite postmilking teat disinfectants with sodium dodecylbenzene sulfonic acid on prevention of contagious mastitis using an experimental challenge protocol. AB - Two acidified sodium chlorite postmilking teat disinfectants were evaluated for efficacy against Staphylococcus aureus and Streptococcus agalactiae by using National Mastitis Council experimental challenge procedures. The effect of these teat dips on teat skin and teat end condition was also determined. Both dips contained 0.32% sodium chlorite, 1.32% lactic, and 2.5% glycerin. Dips differed in the amount of sodium dodecylbenzene sulfonic acid (0.53 or 0.27%) added as a surfactant. Both dips significantly reduced new intramammary infection (IMI) rates compared with undipped controls. The dip containing 0.53% dodecylbenzene sulfonic acid reduced new IMI by Staph. aureus by 72% and Strep. agalactiae by 75%. The dip containing 0.27% dodecylbenzene sulfonic acid reduced new IMI by Staph. aureus by 100% and by Strep. agalactiae by 88%. Changes in teat skin and teat end condition for treatment and control groups varied in parallel over time. Teats treated with either teat dip had higher mean teat skin and teat end scores than control teats at some weeks. However, teat skin and teat end condition did not tend to change from the start to the completion of the trial. Application of the two new postmilking teat dips was effective in reducing new IMI from contagious mastitis pathogens. (Key words: teat dip, contagious mastitis, chlorous acid) PMID- 11860119 TI - Reduction of mastitis caused by experimental challenge with Staphylococcus aureus and Streptococcus agalactiae by use of a quaternary ammonium and halogen-mixture teat dip. AB - A teat-dip formulation containing sodium dichloro isocyanuric acid, bronopol, and quaternary ammonium was tested for efficacy against Staphylococcus aureus and Streptococcus agalactiae intramammary infections (IMI) using an experimental challenge model. Sixty-two Jersey cows from the Hill Farm Research Station (Homer, LA) were used in an 8-wk controlled infection trial to evaluate the teat dip. During the afternoon milking, Monday through Friday for 8 wk, all teats of each cow were immersed to a depth of approximately 25 mm in a challenge suspension containing approximately 5 x 10(7) cfu of Staphylococcus aureus and approximately 5 x 10(7) cfu of Streptococcus agalactiae immediately after milking machines were removed. Immediately after challenge, the distal 25 mm of two contralateral teats were dipped with the experimental teat dip; the remaining two teats served as undipped controls. The experimental teat dip reduced the number of new Staph. aureus IMI by 70.9% and reduced the number of new Strep. agalactiae IMI by 60.0%. Teat end and teat skin condition were characterized as normal and without irritation at the completion of the study. The combination of the three germicides in this experimental teat dip is unique and an effective formulation without adverse effects on condition of teat ends or teat skin. PMID- 11860120 TI - Cloning and localization of the bovine and ovine lysophosphatidic acid acyltransferase (LPAAT) genes that codes for an enzyme involved in triglyceride biosynthesis. AB - Lysophosphatidic acid acyltransferase (LPAAT) catalyzes the addition of fatty acyl moieties to the sn-2 position of the glycerol backbone of lysophosphatidic acid in triglyceride biosynthesis. In this study, we have cloned, sequenced, and characterized the bovine and ovine LPAAT cDNA. Both encode proteins of 287 amino acids with molecular masses of 32 and 31.9 kDa, respectively, differing only by a single amino acid residue. The bovine and ovine LPAAT are predicted to be transmembrane enzymes localized to the endoplasmic reticulum. We also characterized the sequence and genomic organization of the bovine LPAAT gene. The gene consists of seven exons and six introns, spanning a 7.5-kb distance. With the use of a whole genome radiation hybrid panel, we localized the bovine LPAAT to the central region of chromosome 23. PMID- 11860121 TI - Influence of ovine milk in mixture with bovine milk on the quality of reduced fat Muenster-type cheese. AB - Reduced fat Muenster-type cheeses were manufactured from a mixture of bovine skim milk and ovine whole milk and from bovine milk only (control). Cheeses were evaluated at 15, 30, 60, 90, 120, and 180 d of age for numbers and type of microflora, casein hydrolysis, and amounts of free fatty acids. alpha(s1)-Casein degradation was similar for both cheeses during the aging period, but beta-casein degradation proceeded at a faster rate in the control cheese. The total amounts of free fatty acids remained constant throughout the ripening time; however, the cheeses produced with bovine/ovine milk yielded a significantly larger amount of caprylic (C8:0) and capric (C10:0) acids compared with the bovine milk cheeses. Lactobacilli increased during the aging period, while the populations of lactic acid bacteria, yeast and molds, and lipolytic organisms did not increase. Both cheeses had comparable cheese flavor intensity, but the bovine/ovine milk cheese had a greater occurrence of off flavors. The bovine/ovine milk cheeses were firmer than the bovine cheeses throughout the aging period. PMID- 11860122 TI - The effect of ascorbic acid and L-histidine therapy on acute mammary inflammation in dairy cattle. AB - Ascorbic acid and L-histidine were investigated as antioxidant therapies for acute mammary inflammation. Mastitis was induced in eight nonpregnant Holstein cows by intramammary infusion of endotoxin. Treatments were administered in a 4 x 4 Latin square crossover design with 1-wk periods between challenges with endotoxin. Four individual treatments, control, ascorbic acid only, L-histidine only, and ascorbic acid plus L-histidine, were applied. Two doses of 25 g of ascorbic acid administered intravenously at 3- and 5-h postendotoxin challenge increased milk production recovery (9% higher, P < 0.02) and tended to reduce the extent of rumen stasis. Two doses of 25 g of L-histidine similarly administered decreased plasma antioxidant activities 5.5% (P < 0.05). However, ascorbic acid and L-histidine had no effects on rectal temperature, heart rate, respiratory rate, and dry matter intake. The data suggested that ascorbic acid provided some potential benefit for recovery from acute mammary inflammation in dairy cattle. PMID- 11860123 TI - Development and validation of a short-term, serum-free culture system for bovine granulosa cells: evaluation of the effects of somatotropin and growth hormone releasing factor on estradiol production. AB - The objective of this study was to develop and validate a short-term, serum-free culture system to determine whether recombinant bovine somatotropin (rbST) or recombinant bovine growth hormone-releasing factor (rbGRF) altered the estradiol producing capacity of bovine granulosa cells isolated from dominant or subordinate follicles of the first follicular wave. Thus, ovaries were obtained at an abattoir from cows that were between d 2 to 5 or 6 to 10 of the estrous cycle. Three size classes of follicles were isolated from each cow's ovaries: small (2 to 5 mm in diameter), medium (6 to 14 mm), or the largest (6 to 19 mm). In vivo steroid-producing capacity of follicles was assessed by measuring concentration of estradiol, progesterone, androstenedione and 5alpha dihydrotestosterone in each follicle. In vitro steroid-producing capacity was assessed by culturing granulosa cells from the different follicle sizes for 48 h in serum-free media with 19-OH androstenedione and measuring the estradiol and progesterone concentrations in media at the end of culture. The effect of different doses of FSH, rbST, or rbGRF on estradiol and progesterone production by granulosa cells from each follicle size class during d 2 to 5 or 6 to 10 was also evaluated. A high percentage (91.7%) of the largest follicles obtained on d 2 to 5 was estrogen-active (estradiol > progesterone) compared with other follicle classifications (d 2 to 5, small = 0%, medium = 13.8%; d 6 to 10, small = 0%, medium = 3.3%, largest = 33.3%). Estradiol was highest (P < 0.05) in the largest follicle on d 2 to 5 and correlated positively with follicle diameter. The pattern of in vitro production of estradiol by granulosa cells from the different follicle size classes reflected the original in vivo capacity of follicles to produce estradiol. However, only granulosa cells from the largest estrogen-active follicle on d 2 to 5 produced more estradiol than progesterone in vitro. Progesterone production by granulosa cells from all follicle classifications was increased by FSH, but FSH only enhanced estradiol production by granulosa cells from the largest estrogen-active follicles on d 2 to 5. Recombinant bST blocked the FSH-induced increase in estradiol by granulosa cells from the largest estrogen-active follicles on d 2 to 5, whereas rbGRF had no effect on steroid production. Based on these results, we concluded that short term, serum-free culture of bovine granulosal cells obtained from first-wave follicles at an abattoir could be used to reflect reliably the original in vivo estradiol-producing capacity of granulosal cells, and that neither rbST nor rbGRF enhance basal or FSH-induced estradiol production by bovine granulosa cells from first-wave follicles. PMID- 11860124 TI - Growth kinetics and fractal dimensions of casein particles during acidification. AB - Small angle static light scattering was used to study the effect of milk dilution in permeate on the mechanism of acid-induced aggregation of casein particles. Growth kinetics of casein aggregates during acidification was characterized by the succession of four populations of particles. The first one corresponded to casein particles ranging from 0.1 to 1 microm, with a mean value of 0.3 microm. The second population, from 1 to 10 microm, was quickly replaced by a third population, from 10 to 100 microm, which gave rise to the last population measurable, from 100 to 1000 microm. The angular dependence of static light scattering from about 0.01 to 50 degrees was used to determine the fractal dimension (D) of pH-induced casein aggregates. With the formation of about 10 microm aggregates, fractal structures appeared. The D values, determined from double logarithmic plots of intensity versus scatteringvector resulted in values between 1.85 and 2.03. PMID- 11860126 TI - Milk production and economic measures in confinement or pasture systems using seasonally calved Holstein and Jersey cows. AB - This 4-yr study examined total lactation performance of dairy cows in two feeding systems: pasture-based and confinement. Spring and fall calving herds were used and each seasonal herd had 36 cows on pasture and 36 cows in confinement with 282 Holstein and 222 Jersey cows included over seven seasonal replicates. Pasture-fed cows received variable amounts of grain and baled haylage depending upon pasture availability. Confinement cows received a total mixed ration with corn silage as the primary forage. Data were collected on milk production, feed costs, and other costs. Pasture-fed cows produced 11.1% less milk than confinement cows. Across treatments, Jerseys produced 23.3% less milk than Holsteins, but calving season and various interactions were not significant. Feed costs averaged $0.95/cow per day lower for pastured cows than confinement cows. Feed costs were lower for Jerseys than Holsteins and for cows calving in spring. Income over feed costs averaged $7.05 +/- 0.34 for confinement Holsteins, $6.89 +/- 0.34 for pastured Holsteins, $5.68 +/- 0.34 for confinement Jerseys, and $5.36 +/- 0.34 for pastured Jerseys; effects of breed were significant but treatment, season, and interactions were not. Economic factors such as labor for animal care, manure handling, forage management, and cow culling rates favored pastured cows. Higher fertility and lower mastitis among Jerseys partially offsets lower income over feed cost compared with Holsteins. Milk production was lower in this study for pasture-based systems but lower feed costs, lower culling costs, and other economic factors indicate that pasture-based systems can be competitive with confinement systems. PMID- 11860125 TI - Digestibility by dairy cows of monosaccharide constituents in total mixed rations containing citrus pulp. AB - Ten lactating cows were divided into two groups and individually fed ad libitum one of two experimental total mixed rations (TMR) as follows: 1) a TMR containing 20% corn grain and 10% dry citrus pulp (high corn); and 2) a TMR containing 21% citrus pulp and 9% corn grain (high citrus pulp). Both TMR also contained corn silage (28%), legume haylage (19.5%), extruded full fat soybeans (13.5 to 14%), soybean meal (6.5%), and minerals and vitamins. Voluntary dry matter intake of cows consuming the high corn ration tended to be higher than for cows in the high citrus pulp group (22.2 vs. 20.7 +/- 1.0 kg/cow per day, respectively). Digestibility of NDF was higher for the TMR with high citrus pulp compared with the TMR with higher corn. Across treatments, neutral detergent fiber (NDF) glucose was more digestible than NDF xylose and uronic acids and less digestible than NDF arabinose, galactose, and mannose. Digestibility of total neutral detergent soluble carbohydrates was also higher in the high citrus pulp group (86.8, high citrus pulp vs. 80.3 +/- 1.7%, high corn); with glucose and uronic acids accounting for most of that difference. Consequently, higher total carbohydrate digestibility was obtained in the TMR containing higher citrus pulp compared with the TMR with a higher percentage of corn (77.1 and 72.5 +/- 1.0%, respectively). Slightly lower carbohydrate intake in cows fed higher citrus pulp was compensated by higher digestibility of carbohydrates and protein. Thereby, partial replacement of corn by citrus pulp in TMR of high producing dairy cows improved feed efficiency. PMID- 11860127 TI - Analysis of quinolone residues in edible animal products. AB - A comprehensive review on the analysis of quinolone antibacterials is presented. The review covers most of the methods described for the determination of quinolone residues in edible animal products. Sample handling, chromatographic conditions and detection methods have been discussed. A summary of the most relevant information about the analytical procedures has been included. PMID- 11860128 TI - Pore network modelling of the behaviour of a solute in chromatography media: transient and steady-state diffusion properties. AB - A pore network model is presented, that is a geometrical simplification of a porous medium. The network consists of pore chambers interconnected by pore throats. A recursive algorithm for the simulation of mercury intrusion porosimetry in the network is presented. Calculations indicate that it is possible to fit simulated mercury intrusion data to experimental data, and thereby obtain parameters of the pore size distribution and pore topology (pore connectivity). A time-dependent material balance equation for diffusion on the pore level is set up and solved for the pore network. By calculating the concentration evolution in the network, the transient diffusivity and the steady state diffusivity are found. When the network is well connected, those two diffusivities are equal, but for poorly-connected networks they differ. For migrating solutes that are non-negligibly small compared to the pore throats, considerable differences between the transient and steady-state diffusivities were found. PMID- 11860129 TI - Study of feed temperature control of chromatography using computional fluid dynamics simulation. AB - Recently preparative high-performance liquid chromatography (HPLC) has been used more and more frequently to separate drugs and natural substances. However, large scale HPLC easily tends to reduce the yield and purity of the product. Hydrodynamic and heat factors play an important roles. Generally, in a large scale HPLC column, the tracer profile inside column will take on a parabolic shape because of the distributor, which will impact the separation performance of the column. With the inlet temperature suitably lower than the wall temperature, this situation could be improved to some extent. In this work, some experiments were conducted using HPLC, with a column 10 cm in diameter to determine the optimal temperature difference between wall and inlet temperatures. The wall temperature was fixed at about 30 degrees C and the inlet temperature varied from 15 to 30 degrees C. The flow-rate of the eluent, methanol, was 300 ml/min. The experimental result was simulated using CFD software FLUENT 4.4.4. The simulated temperature field fitted the experimental one very well and the simulated flow, temperature and tracer distribution inside column could provide good explanation of separation performance under different conditions. In addition, the simulation could at least approximately predict the optimal temperature difference. PMID- 11860130 TI - Determination of the xenoestrogens 4-nonylphenol and bisphenol A by high performance liquid chromatography and fluorescence detection after derivatisation with dansyl chloride. AB - An easily performable and highly selective method for the determination of the xenoestrogens bisphenol A (BPA) and technical 4-nonylphenol (mixture of isomers) from environmental samples was developed. The method consists of fluorigenic labelling of the substances by dansylation followed by HPLC separation of the derivatives. Specific wavelengths (lembda(ex)=354 nm, lambda(em)=545 nm) for detection of the dansylated phenols were determined in order to reduce the signals of interfering compounds. The applicablility of the method for environmental samples was demonstrated by using sewage sludge spiked with BPA and 4-n-nonylphenol (as internal standard). PMID- 11860131 TI - Memory effect of mobile phase additives in chiral separations on a Chiralpak AD column. AB - Using chiral probes shown to be sensitive to the presence of mobile phase additives, a memory effect for these additives by an amylosic column was demonstrated. Exposure to these additives gave prolonged chromatographic performance changes even after their removal from the mobile phase. This finding is consistent with strong binding of the additives to the stationary phase. A procedure to remove bound additives was developed. PMID- 11860132 TI - Enantioseparation of amino acids on a polysaccharide-based chiral stationary phase. AB - Sulfonic acids have been shown to be more effective than the commonly used trifluoroacetic acid (TFA) in the chiral resolution of underivatized aromatic amino acids on an amylosic column. Sulfonic acid additives give a more UV transparent mobile phase, possibly allowing the detection of non-aromatic analytes. Work presented demonstrates that through the combination of sulfonic acid mobile phase additives, amine mobile phase additives and solvent modifier variations, the enantiomers of 20 of 25 probe amino acids are fully resolved, four are partially resolved with only one failing to be separated on a common amylosic column. PMID- 11860133 TI - Study of the composition of tars produced from blends of coal and polyethylene wastes using high-performance liquid chromatography. AB - Tars produced at semi-industrial scale in a coke oven of 6 x 10(3) kg capacity were used to investigate the effect of using polyethylene waste as an additive in the carbonization process with coal. The polyethylene wastes used were low density polyethylene from the agriculture greenhouses and high-density polyethylene from domestic sources. The high-performance liquid chromatography analysis of the soluble fractions in toluene and carbon disulfide, using two polystyrene-divinylbenzene columns and a mixture of dichloromethane-methanol as a mobile phase, provides useful information on the composition of tars and their derived pitches in terms of the substitution and molecular topology of polynuclear aromatic compounds (PACs). Differences in composition of tars produced with polyethylene waste at 1% (w/w) have been found to be negligible, while a higher amount of the waste (3%, w/w) promoted the formation of peri condensed PACs at the expense of the substituted cata-condensed PACs. This behaviour is due to more extensive secondary reactions of tar precursors via dealkylation and aromatic condensation taking place during the carbonization process as a consequence of a more viscous co-carbonizing system. Changes in tar composition caused by this amount of polyethylene waste addition were comparable to those promoted by an increase in the carbonization temperature at semi industrial and industrial ovens and by the coal preheating before the carbonization process. The characteristic features in tar composition were also found for the derived pitches from tars obtained with the polyethylene waste addition. PMID- 11860134 TI - Use of self-training artificial neural networks in modeling of gas chromatographic relative retention times of a variety of organic compounds. AB - A quantitative structure-activity relationship study based on multiple linear regression (MLR), artificial neural network (ANN), and self-training artificial neural network (STANN) techniques was carried out for the prediction of gas chromatographic relative retention times of 13 different classes of organic compounds. The five descriptors appearing in the selected MLR model are molecular density, Winer number, boiling point, polarizability and square of polarizability. A 5-6-1 ANN and a 5-4-1 STANN were generated using the five descriptors appearing in the MLR model as inputs. Comparison of the standard errors and correlation coefficients shows the superiority of ANN and STANN over the MLR model. This is due to the fact that the retention behaviors of molecules show non-linear characteristics. Inspection of the results of STANN and ANN shows there are few differences between these methods. However, optimization of STANN is much faster and the number of adjustable parameters for this technique is much less compared with those of the conventional ANN. PMID- 11860135 TI - Improving the accuracy of Kovats' retention indices in isothermal gas chromatography. AB - Isothermal Kovats' retention indices are currently reported as whole numbers, and are frequently deduced from a linear least mean squares fitting of the logarithms of adjusted retention times of a number of n-alkanes versus carbon number, following an iterative method that minimises errors. The currently accepted accuracy is about one retention index unit for apolar stationary phases, and lower for polar stationary phases. This paper presents results that show how the accuracy of the retention index may be safely reported to one-tenth of a retention index unit by the use of a non-linear equation, with present day gas chromatographs without electronic flow controllers. Results are presented that prove the correctness of the retention indices found for several substances on one particular capillary column. Hints on the minimum retention times needed to achieve the 0.1 retention index accuracy are mentioned, for retention times recorded in minutes and in seconds. According to results of this paper, two chromatograms, run under the appropriate conditions, are sufficient to obtain the desired accuracy. The method proposed in this paper does not require knowledge of the hold-up time of the chromatogram. PMID- 11860136 TI - Development of a static headspace gas chromatographic procedure for the routine analysis of volatile fatty acids in wastewaters. AB - An optimised procedure has been developed for the routine analysis of volatile fatty acids in wastewater matrices, using static headspace gas chromatography with flame ionisation detection. Factors such as sample volume, sample pre treatment and the time and temperature of sample equilibration have been included in an optimisation model designed to provide maximum detector response for acetic, propionic, iso- and n-butyric and iso- and n-valeric acids in the concentration range 0-1000 mg/l. Optimal headspace conditions were observed when equilibrating at 85 degrees C for 30 min, using a 2.0 ml sample volume with the addition of 1.0 ml of NaHSO4 (62%, w/v) into standard 22.3 ml vials. 2 Ethylbutyric acid was used as an internal standard. The suitability of ordinary least squares regression and weighted least squares regression models for the purposes of calibration and quantification were investigated. A weighted least squares linear regression model applied to the heteroscedastic data provided lower detection limits, e.g. 3.7 and 3.3 mg/l for acetic and propionic acids. PMID- 11860137 TI - Application of headspace solid-phase microextraction to the determination of sulphur compounds with low volatility in wines. AB - Headspace solid-phase microextraction (HS-SPME) has been used for determining sulphur compounds with low volatility in wines. With this technique, handling of samples is minimal so undesirable loses and reactions between compounds are prevented. Furthermore, this kind of extraction is fast and does not require any organic solvent. Under optimal conditions, the HS-SPME, using a new fibre coated with Stable Flex divinylbenzene-Carboxen-polydimethylsiloxane, makes possible the quantification of sixteen sulphur compounds with low volatility which may be present in wines. The limits of detection for the analytes studied ranged between 0.05 and 10 microg/l, and the recovery and repeatability found were acceptable. The method developed was successfully applied to determine the concentration of the target analytes in varietal wines from the Catalonian region (Spain) with some aromatic defects such as an odour of rubber, onion, rotten, unpleasant herbaceous, etc. The results show that the contents of the sulphur compounds studied in these wines are higher than in those without defects. This shows a relationship exists between the presence of sulphur compounds and the quality of the wine aroma. PMID- 11860138 TI - Analysis of phenolic acids by micellar electrokinetic chromatography: application to Echinacea purpurea plant extracts. AB - A micellar electrokinetic chromatographic (MEKC) method was developed for the separation of ten phenolic acids including cichoric acid and caftaric acids, specific marker phytochemicals of Echinacea purpurea. The MEKC method involved the use of 70 mM sodium deoxycholate (SDC) in 40 mM borate (pH 9.2) buffer and UV detection at 300 nm. The bile acid was used as biosurfactant able to provided a micellar system with different and more selective properties than sodium dodecyl sulfate. The effects of SDC and borate concentration and buffer pH on the analyte resolution were evaluated. The validated method was applied to the determination of cichoric acid and related compounds in E. purpurea root extracts, and in commercial E. purpurea based dried extracts and tablets. PMID- 11860139 TI - Chiral separation of underivatized amino acids by ligand-exchange capillary electrophoresis using a copper(II)-L-lysine complex as selector. AB - A ligand-exchange capillary electrophoretic method was explored, with L-lysine as the ligand and copper(II) as the central ion. Its applicability was demonstrated with underivatized aromatic amino acids, namely D,L-phenylalanine, D,L tryptophan, D,L-tyrosine and D,L-beta-phenylserine. Optical resolutions of a single pair of amino acid enantiomers, and of mixed amino acids were obtained with a running buffer of 10 mM NH4Ac, 6.67 mM Cu(II) and 13.33 mM L-lysine, pH 7.0. Sodium dodecylsulfate (SDS) was shown to be necessary for simultaneous separation of the mixed amino acids. The resolution was found to increase with the concentration of the copper(II) complex at a copper(II)-to-lysine ratio of 1:2. If the total concentration of copper(II) and lysine was kept at 20 mM, decreasing the ratio of copper(II) to lysine caused a resolution loss of tryptophan, but a slight resolution improvement of the other three amino acids. The pH of buffer is another important factor controlling the separations. For all the studied amino acids, the optimum pH was 6.0. An interesting phenomenon was observed in this study. SDS induces precipitation at a concentration below 32 mM at room temperature (16+/-2 degrees C), possibly due to the formation of neutral substance from the SDS monomer and the copper(II)-lysine complex. PMID- 11860140 TI - Capillary electrophoresis of platinum-group elements. Analytical, speciation and biochemical studies. AB - A great deal of research has been carried out in recent years on developing high efficiency capillary electrophoresis (CE) techniques that are able to separate rapidly and selectively ionic platinum metal species in a wide variety of their complexed forms. Using a range of illustrative examples, this review examines the potential and utilization of various CE separation approaches and detection modes in this expanding area. Also covered are CE procedures suitable for solving practical analytical problems and for platinum metal speciation purposes. Presenting a comprehensive treatise on the evolving practices of CE concerning platinum anticancer drugs--in particular, the examination of the stability of intact drugs, the separation and identification of products of their metabolism and interactions with biomolecules (including kinetic studies of the binding behavior)--this paper witnesses a welcome shift of the main research activities to those performed under physiological conditions. PMID- 11860141 TI - Analysis of plant hormones in tobacco flowers by micellar electrokinetic capillary chromatography coupled with on-line large volume sample stacking. AB - Micellar electrokinetic capillary chromatography was developed to analyze plant hormones including gibberellic acid, abscisic acid, indole-3-acetic acid, alpha naphthaleneacetic acid, 2,4-dichlorophenoxyacetic acid, kinetin-6 furfurylaminopurine and N6-benzyladenine. The influences of some crucial parameters including buffer concentration, pH value, micelle concentration and applied voltage on electrophoretic separation were investigated. Under optimum conditions (50 mM borate as the running buffer containing 50 mM sodium dodecylsulfate, pH 8.0; separation voltage: -15 kV; injection: hydrodynamic injection, 5 s at 50 mbar; temperature: 25 degrees C), a complete separation of seven plant hormones was accomplished within 30 min. Emphasis was placed on improving detection sensitivity in order to detect small amounts of hormones in plant tissue. Multiple wavelength detection and expanded bubble cell capillary were used with enrichment factors of 2 and 3, respectively. In addition, an on line concentration method of large volume sample stacking was designed. Enrichment factors of up to approximately 10-600 were achieved for these hormones with detection limits down to 0.306 ng/ml. The method was successfully applied to analyzing abscisic acid in flowers of transgenic tobacco. PMID- 11860142 TI - On-column trace enrichment by sequential frontal and elution electrochromatography II. Enhancement of sensitivity by segmented capillaries with z-cell configuration--application to the detection of dilute samples of moderately polar and nonpolar pesticides. AB - An on-column trace enrichment method for capillary electrochromatography of dilute samples is described. It involves the sequential use of frontal and elution electrochromatography on a segmented capillary column comprising of two contiguous segments each packed with a different sorbent. While the entering segment is for preconcentration by frontal electrochromatography the second segment is much longer and is meant for separation of the enriched analytes in the subsequent elution electrochromatography step. The preconcentration segment is usually packed with a sorbent that affords the highest affinity towards the solutes of interest while the separation segment is packed with a stationary phase that exhibits the highest selectivity and separation efficiency for the analytes. The detection is performed in the UV using a z-cell configuration for achieving an increased path length for detection. The effectiveness of this on column trace enrichment is demonstrated on dilute samples of moderately polar solutes (e.g., carbamate insecticides) and nonpolar solutes (e.g., pyrethroid insecticides). Under optimal frontal and elution electrochromatography conditions. 817- and 1100-fold sensitivity increase are achieved for permethrin (a pyrethroid insecticide) and methiocarb (a carbamate insecticide), respectively, with a UV detector. The method is demonstrated with real water samples (e.g., tap and lake water samples) spiked with carbamate and pyrethroid insecticides. The limits of detection for the pesticides achieved in tap and lake waters reached 10(-8) to 10(-9) M. PMID- 11860143 TI - Separation of tanshinones from Salvia miltiorrhiza bunge by multidimensional counter-current chromatography. AB - Analytical and preparative high-speed counter-current chromatography (HSCCC) was successfully used for the isolation and purification of tanshinones from the roots of Salvia miltiorrhiza Bunge. Using multidimensional HSCCC, four major components including tanshinone IIA (16 mg), tanshinone I (10 mg), dihydrotanshinone I (7 mg) and cryptotanshinone (11 mg) were isolated each at high purity of over 95%. PMID- 11860145 TI - Towards ochratoxin A selective molecularly imprinted polymers for solid-phase extraction. AB - Molecularly imprinted polymers (MIPs) displaying selective binding properties for the mycotoxin ochratoxin A (OTA) in polar/protic media were prepared. Crucial to the success of these efforts was the implementation of rationally designed OTA mimics as templates and a set of novel basic and neutral functional monomers, allowing the maximization of the template-functional monomer association via ion pairing, hydrophobic and steric interactions. MIPs prepared with a 20:1:1:3 molar ratio of cross-linking agent, template mimic, basic functional monomer and hydrophobic auxiliary monomer produced polymers with superior recognition properties compared to materials generated with other stoichiometries. Chromatographic evaluation using the OTA mimics, OTA and a set of structurally closely related compounds as analytes revealed pronounced substrate selectivity of these MIPs in polar/protic media, the templates and OTA being bound with significantly higher affinities. Complementary substrate selectivities/affinities were observed in aprotic and apolar solvents. The possibility of solvent dependent tuning of substrate selectivity/affinity and the high binding capacity recommend the developed MIPs as promising solid-phase extraction adsorbents for clean-up and pre-concentration of OTA from various biologically relevant matrices. PMID- 11860146 TI - Axial development and radial non-uniformity of flow in packed columns. AB - Flow inhomogeneity and axial development in low-pressure chromatographic columns have been studied by magnetic resonance imaging velocimetry. The columns studied included (a) an 11.7-mm I.D. column packed with either 50 microm diameter porous polyacrylamide, or 99 or 780 microm diameter impermeable polystyrene beads, and (b) a 5-mm I.D. column commercially packed with 10 microm polymeric beads. The packing methods included gravity settling, slurry packing, ultrasonication, and dry packing with vibration. The magnetic resonance method used averaged apparent fluid velocity over both column cross-sections and fluid displacements greater than one particle diameter and hence permits assessment of macroscopic flow non uniformities. The results confirm that now non-uniformities induced by the conical distributor of the 11.7-mm I.D. column or the presence of voids at the column entrance relax on a length scale of the column radius. All of the 11.7-mm I.D. columns examined exhibit near wall channeling within a few particle diameters of the wall. The origins of this behavior are demonstrated by imaging of the radial dependence of the local porosity for a column packed with 780 microm beads. Columns packed with the 99-microm beads exhibit reduced flow in a region extending from ten to three-to-five particle diameters from the wall. This velocity reduction is consistent with a reduced porosity of 0.35 in this region as compared to approximately 0.43 in the bulk of the column. Ultrasonicated and dry-packed columns exhibit enhanced flow in a region located between approximately eight and 20 particle diameters from the wall. This enhancement maybe caused by packing density inhomogeneity and/or particle size segregation caused by vibration during the packing process. No significant non-uniformities on length scales of 20 microm or greater were observed in the commercially packed column packed with 10 microm particles. PMID- 11860144 TI - Simultaneous reversed-phase high-performance liquid chromatographic separation of mono-, di-and trichloroanilines through a gradient elution optimised by experimental design. AB - A new RP-HPLC method for the simultaneous determination of the 13 mono-, di- and trichloroanilines has been developed. In order to obtain the analyte resolution within an acceptable analysis time, a gradient elution program has been optimised through the use of an experimental design and a grid search algorithm. The optimized conditions provided the resolution of all the analytes in less than 80 min. The primary validation of the analytical method gave limit of detection values ranging between 0.02 and 0.06 mg/l and very good linearity of the calibration curves. PMID- 11860147 TI - Retention of ionizable compounds on high-performance liquid chromatography XI. Global linear solvation energy relationships for neutral and ionizable compounds. AB - A global linear solvation energy relationship (LSER) that simultaneously models retention in reversed-phase liquid chromatography as a function of solute LSER descriptors and mobile phase pH and composition has been derived from both the local LSER model and the linear solvent strength theory. At most only 13 mobile phase parameters and seven solute parameters are required to establish the global LSER model for neutral and ionizable solutes. This model implies only one mobile phase and two solute parameters more than the model previously set for neutral solutes. The additional mobile phase and solute parameters account for the ionization of the solute. The model has been successfully tested for 30 solutes of different type (acids, bases and non ionizable compounds) at 10 different pH values in three different acetonitrile-water mobile phases. PMID- 11860148 TI - Nonlinear function retention model in weak acid anion chromatography. AB - A nonlinear function retention model for anion chromatography is developed, which is able to predict the retention behavior of polyvalent weak acid anions using sodium hydroxide as eluent. The relationship between the retention factors of phosphate and resorcinol and eluent concentration was studied with sodium hydroxide as the eluent. Both retention factors of the analyte anions first increase then decrease with the increase of sodium hydroxide concentration. The estimated values agree with the measured values with correlation coefficients for phosphate and resorcinol of 0.9920 and 0.9916, respectively, which shows the nonlinear function model is a useful addition to the theory of anion chromatography. The model is able to optimize the separation of polyvalent weak acid anions. PMID- 11860149 TI - An analysis of the mechanism by which cetiedil inhibits sickling. AB - A study has been carried out into the effects of cetiedil on the activities of Na+, K+ and Ca2+, Mg2+-ATPases of the normal human erythrocyte membrane. In general, cetiedil inhibits both ATPases activities but with characteristic inhibition profiles and varying degrees of efficacy. The activities were inhibited non-competitively at the cetiedil concentration which caused 50% inhibition of each enzyme. In addition, the effects of cetiedil on the transport of K+ and phosphate ions across the membrane were monitored and compared. Cetiedil was found to stimulate K+ release and to inhibit phosphate uptake. At low concentrations, both processes were concentration dependent. Stimulation of K+ efflux reached a plateau at a concentration of 1.2 mM. The antisickling effect of cetiedil is explained mainly in the light of the changes it induces in the activities of membrane-bound ATPases and the permeability properties of the erythrocyte membrane to cations and anions. PMID- 11860150 TI - Corticotropin-releasing hormone affects cytokine production in human HaCaT keratinocytes. AB - CRH cutaneous expression is significantly enhanced after exposure to various stimuli (Physiol Rev 2000, 80;979-1020). We evaluated the effect of CRH on cytokine production in HaCaT keratinocytes, a cell line shown to express CRH receptors coupled to cAMP activation and calcium-dependent transmission pathways. It is demonstrated for the first time that exogenously added CRH stimulates production of IL-6 and IL-11. It also inhibits production of IL-1beta and does not affect TNF-alpha production. Our results indicate that CRH function(s) during cutaneous stress may be mediated by differential effects on cytokine production. PMID- 11860151 TI - Antioxidant activity of anti-inflammatory plant extracts. AB - The antioxidant properties of twenty medical herbs used in the traditional Mediterranean and Chinese medicine were studied. Extracts from Forsythia suspensa, Helichrysum italicum, Scrophularia auriculata, Inula viscosa, Coptis chinensis, Poria cocos and Scutellaria baicalensis had previously shown anti inflammatory activity in different experimental models. Using free radical generating systems H. italicum. I. viscosa and F. suspensa protected against enzymatic and non-enzymatic lipid peroxidation in model membranes and also showed scavenging property on the superoxide radical. All extracts were assayed at a concentration of 100 microg/ml. Most of the extracts were weak scavengers of the hydroxyl radical and C. chinensis and P. cocos exhibited the highest scavenging activity. Although S. baicalensis inhibited the lipid peroxidation in rat liver microsomes and red blood cells, the extract showed inhibitory actions on aminopyrine N-demethylase and xanthine oxidase activities as well as an pro oxidant effect observed in the Fe3+-EDTA-H2O2 system. The results of the present work suggest that the anti-inflammatory activities of the same extracts could be explained, at least in part, by their antioxidant properties. PMID- 11860152 TI - Comparison of the total antioxidant content of 30 widely used medicinal plants of New Mexico. AB - Teas made from medicinal plants are commonly used by a majority of the inhabitants of New Mexico and Mexico to treat various ailments including infections, arthritis, heart disorders, headaches, fever, asthma and menstrual pain. However, little is known about the identity or chemical nature of the bioactive substances and compounds responsible for the therapeutic effects of the teas made from the leaves, seeds, flowers stems, and roots of these medicinal plants. Some of the beneficial effects of these teas may be attributable to antioxidants contained in the medicinal plants from which they are brewed. In the present study we collected 30 medicinal plants that are widely used in the Rio Grande Valley and, using a two-stage Trolox based assay, analyzed the total antioxidant capacity of aqueous extracts prepared from these plants. The antioxidant content of the aqueous extracts was substantial, ranging from 27 to 972 micromol Trolox equivalent per gram dry weight. An extract of the leaves of the plant Ilex paraguensis (Mate leaf) contained the highest amount of antioxidant, followed by the flowers of the Rosa sp. (Rosa de Castillo, 804 micromol/g), the bark of Chinchona sp. (Copalquin, 692 micromol/g), Rumex hymenosepalus stems (Cana Agria, 672 micromol/g) and the leaves of Marrubium vulgare (Mastranzo, 560 micromol/g). The plants that had the lowest antioxidant capacity were the seeds of Linum lewissii (Linasa, 29 micromol/g) and Yucca sp. plant root (Amole, 27 micromol/g). It will be useful to further analyze those plants that contain the most antioxidant activity in order to identify the active principles. PMID- 11860153 TI - Assessment of the relationship between hyperalgesia and peripheral inflammation in magnesium-deficient rats. AB - Magnesium-deficient rats develop simultaneously a significant lowering of nociceptive threshold and a generalized inflammation. We investigated the relationship between these two phenomena by testing drugs that are able to suppress the inflammation in this model. In weaning rats fed a magnesium-depleted diet for ten days, the nociceptive threshold was assessed by the paw pressure test and the inflammation by a clinical score. A non-steroidal anti-inflammatory drug (piroxicam); antagonists of H1 and H2 receptors (astemizole and cimetidine. respectively); a glucocorticoid (dexamethasone); an inhibitor of mastocyte degranulation (cromoglycate); and estradiol benzoate were used to block the inflammatory response. Dexamethasone and estradiol significantly suppressed the inflammation (p < 0.001 vs control group). Cromoglycate showed a delayed anti inflammatory effect (p < 0.01 vs control group on D10). The combination of astemizole and cimetidine partially blocked the inflammation process, whereas astemizole and piroxicam were without effect. Regardless of the effect of the test drugs on inflammation, no change in the time course of hyperalgesia was observed. These data support the view that hyperalgesia induced by the magnesium depleted diet is not a consequence of the inflammatory process. PMID- 11860154 TI - Nociceptin/orphanin FQ increases ANP secretion in neonatal cardiac myocytes. AB - Nociceptin (N/OFQ) is a novel heptadecapeptide with an amino acid sequence similar to that of endogenous opioid peptide dynorphin A. Dynorphin have been reported to increase the secretion of atrial natriuretic peptide (ANP) via selective activation of kappa-opioid receptor in cultured atrial cardiocytes. The present study was designed to investigate the direct effect of N/OFQ on the ANP secretion in cultured neonatal rat cardiac myocytes via N/OFQ receptor (NOP) activation. The secretion of ANP from cultured neonatal cardiac myocytes was increased in terms of incubation time. N/OFQ, at a dose of 0.3, 1, 3, and 10 microM, caused increases in ANP secretion in a dose-dependent manner. The N/OFQ induced ANP secretion was completely antagonized by antagonists of NOP, 1 microM each of [Phe1 (CH2-NH) Gly2] nociceptin (1-13)-NH2 ([FG]N/OFQ(1-13)NH2) or naloxone benzoylhydrazone. In contrast, naloxone (1 microM), the non-selective opioid receptor antagonist, did not alter ANP response to N/OFQ. N/OFQ at 3 microM inhibited basal and forskolin-stimulated cAMP production, which was partially antagonized with the pretreatment of [FG]N/OFQ(1-13)NH2. An increase in ANP secretion by N/OFQ was also partially blocked by the pretreatment of forskolin. Homologous competition studies in neonatal cardiomyocyte membranes revealed the presence of two distinct sites. The high affinity site (10.9 +/- 1.6 nM) was far less abundant than the low affinity site. Therefore, these results suggest that N/OFQ causes an increase in ANP secretion in cultured neonatal cardiac myocytes by decreasing cAMP through its binding sites. PMID- 11860155 TI - Isolation of lilin, a novel arginine- and glutamate-rich protein with potent antifungal and mitogenic activities from lily bulbs. AB - From the dried bulbs of the lily (Lilium brownii), a protein with strong antifungal and mitogenic activities was isolated. It also exhibited an inhibitory action on the activity of HIV-1 reverse transcriptase. The protein was single chained and possessed a molecular weight of 14.4 kDa and an N-terminal sequence distinct from chitinases and antimicrobial proteins of garlic, leek and onion which belong to a family closely related to lily. However, there was a small degree of resemblance to cyclophilins and a considerable extent of identity to the 6.5 kDa arginine/glutamate-rich polypeptide from Luffa cylindrica seeds. A nearly homogeneous preparation was obtained after the extract was fractionated on DEAE-cellulose and Affi-gel Blue gel since subsequent chromatography on Mono S and Superdex 75 both yielded a single peak. PMID- 11860156 TI - Nociceptin and dynorphin A(1-17) produce antianalgesia through independent systems in mice. AB - The administration of dynorphin A(1-17), Dyn, intrathecally (i.t.) or of nociceptin, intracerebroventricularly (i.c.v.) produces antianalgesic actions against i.t. morphine in the tail flick test in mice. The antianalgesic action of nociceptin is mediated by spinal PGE2 and attenuated by i.t. PGD2 or indomethacin. The Dyn response is mediated by release of IL1beta in the spinal cord to activate an ascending pathway to the brain and in turn releases IL1beta in the brain which activates a descending pathway to the spinal cord. The present work investigated the possibility that the action of IL1beta in the Dyn system might release prostaglandins so that the Dyn and nociceptin antianalgesic systems would overlap at these points. The results indicated that in the Dyn system neither the IL1beta in the spinal cord or brain implicated prostaglandin release because i.t. and i.c.v. PGD2 and indomethacin did not affect Dyn-induced antianalgesia. In addition, nociceptin-induced antianalgesia did not involve components in the Dyn system. Thus, the Dyn and nociceptin antianalgesic systems did not overlap and each were independent systems. PMID- 11860157 TI - Effects of short- and long-acting dopamine agonists on sensitized dopaminergic neurotransmission in rats with unilateral 6-OHDA lesions. AB - The effects of short and long-acting dopamine agonists on sensitized dopaminergic transmission in an animal model of Parkinson's disease were investigated. Rats with 6-hydroxydopamine (6-OHDA) lesions of the left nigrostriatal dopaminergic pathway were pre-exposed i.p. to 50 mg/kg methyl levodopa for 10 days. After a 7 day withdrawal period, these animals were treated with saline i.p., 0.05 mg/kg apomorphine s.c., or 0.5 mg/kg cabergoline i.p., once daily for 7 days. On the 8th day, rats in each treatment group received a challenge dose of 0.05 mg/kg apomorphine or saline s.c. The temporal changes in the number of rotations away from the 6-OHDA lesion side were evaluated after the challenge. The apomorphine challenge increased the number of rotations more markedly in the apomorphine pretreated rats than in the other pretreatment groups. In cabergoline pretreated rats, the number of rotations was significantly lower than that of saline pretreated animals. Pretreatment with saline did not alter the apomorphine sensitivity of rotational behavior. These findings suggest that the repeated administration of long-acting dopamine agonists may reduce sensitized dopaminergic transmission in dopamine-depleted rats, whereas short-acting ones may further enhance sensitization of the transmission process. PMID- 11860158 TI - Differences between D- and L-aspartate binding to the Na+-dependent binding sites on glutamate transporters in frozen sections of rat brain. AB - The Na+-dependent, "high-affinity" transport of L-glutamate (GluT) in brain tissue has become a significant focus of interest, particularly since it has been revealed that abnormalities of GluT may be associated with serious neurological disorders. Using quantitative autoradiography on 3H-sensitive films, we have studied, in thaw-mounted sections of rat brain, the distribution and pharmacology of radioligand binding to sites with characteristics of the substrate recognition/binding locus on GluT. The technique makes it possible to determine not only the intensity of binding in brain regions but, with a high level of precision, pharmacological constants such as IC50 or nH. [3H]L-aspartate and [3H]D-aspartate are two classical radioligands used in studies of GluT. We have determined IC50 values for the inhibition of [3H]L- and [3H]D-aspartate binding by their non-radioactive counterparts in the cerebral neocortex. hippocampus, striatum, septal nuclei and the cerebellar cortex. The two radioligands did not appreciably differ from each other in their interactions with the binding sites in the forebrain, consistent with all Na+-dependent GluT binding sites in that region having no stereoselectivity for aspartate enantiomers. In the cerebellar cortex, however, the data indicated the presence of a GluT binding site that preferred L- over D-aspartate. These findings contrast with many previous observations and suggest that the pharmacological characteristics of the ligand binding sites on GluT in the mammalian cerebellar cortex may have to be re assessed and/or a possibility of an existence of (a) hitherto unknown molecule(s) with properties of a glutamate transporter be considered. PMID- 11860159 TI - The relationship between parental psychiatric disorder and child physical and sexual abuse: findings from the Ontario Health Supplement. AB - OBJECTIVE: The study's goal was to examine the relationship between a history of parental psychiatric disorder and a history of child abuse in a general population sample of Ontario residents. METHOD: A representative community sample of 8548 respondents who participated in the Ontario Mental Health Supplement (OHSUP) were interviewed about parental psychiatric history and completed a self report measure of childhood physical and sexual abuse. RESULTS: The lifetime prevalence of either parent with a psychiatric disorder was as follows: 14.1% for depression, 3.7% for manic depression, 2.4% for schizophrenia, 2.4% for antisocial behavior, and 17.3% for any parental psychiatric disorder. Respondents reporting a parental history of depression, mania, or schizophrenia had a two to threefold increase in the rates of physical, sexual, or any abuse. Parental history of antisocial disorder increased the risk of exposure to physical abuse (adjusted odds ratios [OR 6.1] and any abuse [OR 7.5]). There was no statistically significant difference between parental psychiatric disorder and childhood physical or sexual abuse by gender of the respondent. There was a trend for increasing risk associated with father only, mother only, and both parents having any psychiatric disorder. CONCLUSIONS: The elevated risk for physical and sexual abuse among respondents reporting a parental history of psychiatric illness highlights the need to examine the mechanism for this association. Such information is important in developing approaches to assist families where the risk of child maltreatment is increased. PMID- 11860160 TI - Childhood experience of sexual abuse among Hong Kong Chinese college students. AB - OBJECTIVES: This study aimed to explore the prevalence rate and profiles of abusers and victims of child sexual abuse in a Chinese society. METHOD: A convenience sample of 2,147 Hong Kong Chinese college students completed questionnaires which included items on definitions of child sexual abuse, recall of sexual abuse experience before age 17, information about the sexual abuse incidents, and personal demographics. RESULTS: The overall prevalence rate for various forms of child sexual abuse was 6%, with the rates being higher for females than for males. The average age of the victims at the time of the sexual abuse was 11 years old, with a majority of them experiencing sexual abuse during their teenage years. The average age of the abusers as estimated by the victims was 24 years old, and only 28% of the abusers were strangers to the victims. Males were about three times more likely than females to be sexually abused by their friends, whereas females were about twice more likely than males to be sexually abused by strangers. Only 39% of the victims reported their sexual victimization to others, and 56% of the reported incidents were not followed up. CONCLUSIONS: As compared to Western studies, the present study showed lower rates for more intimate or severe forms of child sexual abuse. The Chinese victims were more reluctant to disclose their sexual victimization than their Western counterparts. Limitations and practical implications of the study are also discussed. PMID- 11860161 TI - Undefined use of the terms "child sexual touching" and "child sexual contact". PMID- 11860162 TI - Differential symptom pattern of post-traumatic stress disorder (PTSD) in maltreated children with and without concurrent depression. AB - OBJECTIVE: The present study attempted to examine specific differences in the Post-Traumatic Stress Disorder (PTSD) symptomatology among abused children with and without concurrent depression. METHOD: PTSD and depressive symptoms were identified that discriminate between 98 children divided into three groups: (1) abused children with PTSD, (2) nonabused children who meet criteria for Major Depressive Disorder (MDD), and (3) abused children with both PTSD and MDD. RESULTS: Analyses revealed that nine items reflecting depressive symptomatology, primarily vegetative symptoms, differentiated the diagnostic groups (PTSD-only, MDD-only, and the combined group). A discriminant analysis revealed that the sum of responses to the nine significant items adequately predicted diagnostic classification for those with PTSD and depression, but did not correctly diagnose any in the combined group. Analyses also revealed that three post-trauma symptoms, including psychological amnesia, flashbacks/reenactments, and sleep difficulties, discriminated between the groups. The PTSD-only group reported more episodes of psychological amnesia while the PTSD and MDD group experienced more flashbacks. CONCLUSIONS: For the sample of abused children examined, these results illuminate differences with respect to PTSD symptom presentation for those children with PTSD who have a concurrent depressive disorder and their nondepressed counterparts. Children with PTSD who have a concurrent depression report greater levels of intrusive PTSD-related symptoms. PMID- 11860163 TI - The perpetrator's strategy as a crucial variable: a representative study of sexual abuse of girls and its sequelae in Switzerland. AB - OBJECTIVE: Little is known about the long-term consequences of "soft" perpetrator strategies. The purpose of the present study is to assess the consequences of love-bargaining strategies compared to those involving force. METHOD: A representative sample of 980 women, aged 20 to 40 years, in the German-speaking part of Switzerland was asked questions regarding psychosomatic well-being, sexual actions, and perpetrator strategies, as well as motivations for tolerating long periods of abuse. The answers regarding strategies were factor analyzed. The effect on psychosomatic well-being was determined by means of stepwise regression analysis and correlation analysis. RESULTS: The prevalence of child sexual abuse (CSA) was 39.8% overall and 14.7% for severe abuse. Using the three variables love-bargaining strategy, self-blaming, and need for affection, an adjusted R2 of .44 was reached in the 3rd step as a dependent variable with a General Symptomatic Index (GSI) calculated from an abbreviated version of the SCL-90-R. The first of the three variables mentioned was by far the most influential. Force was not included in the equation but was closely correlated on a bivariate level with the SCL-90 partial scale "anxiety." CONCLUSIONS: The consequences of CSA produced by the use of soft strategies have been underestimated in the past. PMID- 11860164 TI - Parents with substance use problems and their infants. PMID- 11860165 TI - The risk of subsequent maltreatment allegations in families with substance exposed infants. AB - OBJECTIVE: This study seeks to: (1) assess the relationship between identified prenatal substance use and the risk of subsequent maltreatment allegations among families involved with child protective services; and (2) compare the types of safety threats encountered by children whose parents had substance-exposed infant (SEI) allegations to the types of safety threats faced by children whose parents had other types of allegations. METHOD: Survey data from a probability sample of parents were linked to state administrative data over a 33-month time frame. Cox regression models were conducted to assess the relative risk of subsequent allegations associated with parents whose child welfare case opened following an SEI allegation (the SEI group) compared to parents whose case opened following other types of allegations. RESULTS: The likelihood of subsequent allegations is greater among parents in the SEI group. However, the increased risk stems almost entirely from subsequent SEI-related allegations. Parents in the SEI group are not more likely to incur other types of allegations such as physical abuse or lack of supervision. CONCLUSIONS: An increased risk of subsequent maltreatment has been used to justify opening child protective cases on the basis of an SEI allegation alone. By looking closely at the types of subsequent allegations as well as the incidence of subsequent allegations, this research helps to clarify the maltreatment risks associated with SEI cases. PMID- 11860166 TI - Symptomatic asthma: attendance and prescribing in general practice. AB - Under-prescribing and low attendance continue to be cited as reasons for ongoing asthma symptoms in primary care despite marked increases in prescribing and structured care for asthma over the past 10 years. The objective of this study was to determine the relationship between continuing asthma morbidity and the attendance of and prescribing for symptomatic asthmatic patients in primary care. A random sample of 402 subjects from 801 who reported at least one of six symptoms in the previous month on most or every day were identified from responses to a validated morbidity questionnaire. An analysis of their care over a 2-year period (1 year before and 1 year after the questionnaire) was carried out from their general practice case-notes. Data on 308 patients was available for analysis. Ninety-four per cent of these symptomatic asthma patients attended over the 2-year period, with 77% attending for an asthma related consultation. Most patients were managed exclusively in primary care. Inhaled steroids were prescribed for 78% of patients and high dose inhaled steroids (> or = 800 mcg of beclomethasone or equivalent per day) were prescribed for 38%. Patients with most symptoms were more likely to be prescribed inhaled steroids. Rescue courses of oral steroids were prescribed for 29% of patients. Changes in asthma medications were recorded for 31% during the study period. Metered dose inhalers (MDI) were prescribed for 86% with more than half prescribed MDIs combined with some other delivery device. Elements of structured care were more frequently recorded in patients who reported most symptoms. In conclusion the asthma management of the majority of patients in this study was active with high levels of steroid prescribing. There appeared to be room to increase prescribing and to improve the structure of care. While patients who were 'symptomatic on steroids' should have had their medications, delivery devices and structured care reviewed regularly many were already on maximal treatment and were therefore likely to remain symptomatic. It is unclear how practitioners could improve morbidity in many of these patients as under-treatment and low attendance seem unlikely to be the principal causes of continuing symptoms. PMID- 11860167 TI - Urinary desmosine excretion in acute exacerbations of COPD: a preliminary report. AB - Desmosine (DES) is an elastin-derived, cross-link amino acid, which is not metabolized; hence, its urinary levels reflect elastin breakdown. We hypothesized that elastin degradation should increase as a result of increased lung inflammation during an acute exacerbation of COPD and should decrease after recovery. To test this hypothesis we measured DES in three urine samples from nine COPD subjects during the first 5 days of an acute exacerbation and at 2 months after recovery. We also measured forced expiratory volume in 1 sec (FEV1) to monitor the effects ofthe exacerbation on ventilatory function. The mean (SD) FEV1 was 45 (15)% predicted during the exacerbation and 57.8 (16)% predicted 2 months later (P=0.00001). The mean (SD) DES excretion was 25.3 (9) microg g(-1) creatinine at day 1;23.5 (9) at day 3 and 24 (9) at day 5 of the exacerbation. The mean (SD) urinary DES excretion 60 days after discharge was 20.9 (7) microg g(-1) creatinine (P=0.049) in comparison with the mean of the three acute-phase values. The size of the increase in desmosine excretion during exacerbation is small, 3.2 microg g(-1) creatinine or 16% of the recovery desmosine value. We conclude that there is a small but statistically significant increase in lung elastin breakdown in the body during an acute exacerbation of COPD. PMID- 11860168 TI - Exhaled breath condensate and serum levels of hepatocyte growth factor in pneumonia. AB - Hepatocyte growth factor (HGF) is a protein produced by mesenchymal cells in many organs, which can stimulate epithelial growth. An enhanced production and concentration of HGF is observed after injuries. The lung is one of the major sources of HGF. By cooling exhaled air, a condensate is formed containing molecules from bronchi and alveoli. In order to investigate HGF-concentration and time course in pneumonia, paired serum and exhaled breath condensate was collected from 10 patients with pneumonia, 10 patients with non-respiratory infections and 11 healthy controls. The concentration of HGF was measured by an immunoassay kit. In the acute phase HGF-levels in breath condensate and serum were significantly higher in the patients with pneumonia compared to the control groups. Similar concentrations in breath condensate were seen in healthy controls and in patients with non-respiratory infections. In the patients with pneumonia a decrease in serum HGF was seen already after 4-7 days while HGF values in breath condensate remained elevated even after 4-6 weeks. These results might imply local product on of HGF in the lungs and a long repair and healing process after pneumonia. PMID- 11860169 TI - Blood eosinophil counts for the prediction of the severity of exercise-induced bronchospasm in asthma. AB - It has been suggested that airway eosinophilic inflammation is associated with the severity of exercise-induced bronchospasm (EIB). Blood eosinophils are known to be an indirect marker of airway inflammation in asthma. The aim of this study is to investigate that a simple and easy blood test for blood eosinphil counts may predict the severity of EIB in asthma. Seventy-seven men with perennial asthma (age range 18-23 years) were included. Lung function test, skin prick test, and blood tests for eosinophils counts and total IgE levels were performed. Methacholine bronchial provocation test and, 24 h later, free running test were carried out. EIB was defined as a 15% reduction or more in post-exercise FEV1 compared with pre-exercise FEV1 value. Atopy score was defined as a sum of mean wheal diameters to allergens. EIB was observed in 60 (78%) of 77 subjects. Asthmatics with EIB showed significantly increased percentages of eosinophils (P<0.01), log eosinophil counts (P<0.001), and atopy scores (P<0.05) and decreased log PC20 values (P < 0.05) compared with asthmatics without EIB. Asthmatics with eosinophils of > 700 microl(-1) (36.9 +/- 12.7%) had significantly greater maximal % fall in FEV1 after exercise than asthmatics with eosinophils of < 350 microl(-1) (24.7 +/- 16.6%, P <0.05). Blood eosinophil counts > 350 microl(-1) yielded the specificity of 88% and positive predictive value of 93% for the presence of EIB. When a multiple regression analysis of maximal % fall in FEV1 according to log eosinophil counts, log PC20, log IgE and atopy score was performed, only blood eosinophil counts were significant factor contributing to the maximal % fall in FEV1 after exercise. These findings not only suggest that a simple blood test for eosinophils may be useful in the prediction of the severity of EIB, but also reinforce the view that airway eosinophilic inflammation may play a major role in EIB in asthma. PMID- 11860170 TI - Re: Elevated level of circulating matrix metalloproteinase-9 in patients with lung cancer (Respir Med 2001; 95: 1-4). PMID- 11860171 TI - Universally low spirometry values in respiratory outpatients. PMID- 11860172 TI - Re: Evaluation of an inspiratory muscle trainer in healthy humans (Respir Med 2001; 95: 526-531). AB - The main outcome of this study is that twitch Pdi is unsuitable for assessing outcmes in small studies of IMT. PMID- 11860173 TI - Expression of MUC5AC and MUC5B mucins in normal and cystic fibrosis lung. AB - Hypersecretion of airway mucus is a characteristic feature of chronic airway diseases like cystic fibrosis (CF) and leads via impairment of the muco-ciliary clearance and bacterial superinfection to respiratory failure. The major components of the mucus matrix forming family of mucins in the airways are MUC5AC and MUC5B. To investigate the expression of these glycoproteins in CF, immunohistochemistry was carried out on trachea, bronchi and peripheral lung obtained from CF patients and compared to normal lung tissues. MUC5AC immunohistochemistry demonstrated signals in goblet cells of the epithelial lining. Also, goblet cells inside glandular secretory ducts revealed MUC5AC positive staining. In comparison to those from normal subjects, CF sections were characterized by inflammatory changes and goblet cell hyperplasia, resulting in increased numbers of MUC5AC-positive cells. Immunohistochemical staining for MUC5B showed abundant staining of submucosal glands and epithelial goblet cells. Inside the glands, the immunoreactivity was restricted to glandular mucous cells. MUC5AC and MUC5B are expressed in the same histological pattern in CF compared to normal tissues with an increase of MUC5AC-positive cells due to goblet cell hyper and metaplasia. PMID- 11860174 TI - Peripheral decrease and pulmonary homing of CD4+CD45RO+ helper memory T cells in cystic fibrosis. AB - Interstitial lung disease, although of prognostic impact for patients with cystic fibrosis (CF), remains difficult to assess without histopathologic investigations. As changes of peripheral blood lymphocyte subsets (LS) may accompany severe systemic lymphocyte immune responses, we compared peripheral LS of 44 patients with CF, 23 non-CF patients with recurrent pulmonary infections and 83 healthy controls (flow cytometry; CD3, CD19, CD16, CD56, CD4, CD8, CD11b, CD45RA, CD45RO, HLA-DR and CD25 antigens). Additional immunohistochemistry was performed on lung tissue of four CF patients aged 0.5, 12, 17 and 20 years, respectively. Patients with CF showed low absolute counts of CD4+CD45RO+ memory helperT cells, CD16+CD56+ NK cells, CD8+ and interleukin-2 receptor-positive T cells in peripheral blood (P < 0.001). Similar changes were registered in the non CF patients with pulmonary infections, indicating that those were not specific for CF. Immunohistochemistry showed activation of bronchus-associated lymphoid tissue with interstitial accumulation of CD4+CD45 RO+ T cells in the three older patients. Patients with CF show marked changes of peripheral blood LS which are presumably not CF-specific and may mirror homing to lung tissue in the course of interstitial lung disease. Further research should evaluate its usefulness in monitoring progression of lung disease in CF. PMID- 11860175 TI - The NADH: ubiquinone oxidoreductase (complex I) of the mammalian respiratory chain and the cAMP cascade. AB - Recent work has revealed cAMP-dependent phosphorylation of the 18-kDa IP subunit of the mammalian complex I of the respiratory chain, encoded by the nuclear NDUFS4 gene (chromosome 5). Phosphorylation of this protein has been shown to take place in fibroblast cultures in vivo, as well as in isolated mitochondria, which in addition to the cytosol also contain, in the inner-membrane matrix fraction, a cAMP-dependent protein kinase. Mitochondria appear to have a Ca2+ inhibited phosphatase, which dephosphorylates the 18-kDa phosphoprotein. In fibroblast and myoblast cultures cAMP-dependent phosphorylation of the 18-kDa protein is associated with potent stimulation of complex I and overall respiratory activity with NAD-linked substrates. Mutations in the human NDUFS4 gene have been found, which in the homozygous state are associated with deficiency of complex I and fatal neurological syndrome. In one case consisting of a 5 bp duplication, which destroyed the phosphorylation site, cAMP-dependent activation of complex I was abolished in the patient's fibroblast cultures. In another case consisting of a nonsense mutation, leading to termination of the protein after only 14 residues of the putative mitochondria targeting peptide, a defect in the assembly of complex I was found in fibroblast cultures. PMID- 11860176 TI - Steady-state kinetics of NADH:coenzyme Q oxidoreductase isolated from bovine heart mitochondria. AB - Steady-state kinetics of the bovine heart NADH:coenzyme Q oxidoreductase reaction were analyzed in the presence of various concentrations of NADH and coenzyme Q with one isoprenoid unit (Q1). Product inhibitions by NAD+ and reduced coenzyme Q1 were also determined. These results show an ordered sequential mechanism in which the order of substrate binding and product release is Q1-NADH-NAD+-Q1H2. It has been widely accepted that the NADH binding site is likely to be on the top of a large extramembrane portion protruding to the matrix space while the Q1 binding site is near the transmembrane moiety. The rigorous controls for substrate binding and product release are indicative of a strong, long range interaction between NADH and Q1 binding sites. PMID- 11860178 TI - Interactions between the cytochrome pathway and the alternative oxidase in isolated Acanthamoeba castellanii mitochondria. AB - The steady-state activity of the two quinol-oxidizing pathways of Acanthamoeba castellanii mitochondria, the phosphorylating cytochrome pathway (i.e. the benzohydroxamate(BHAM)-resistant respiration in state 3) and the alternative oxidase (i.e. the KCN-resistant respiration), is shown to be fixed by ubiquinone (Q) pool redox state independently of the reducing substrate (succinate or exogenous reduced nicotinamide adenine dinucleotide (NADH)), indicating that the active Q pool is homogenous. For both pathways, activity increases with the Q reduction level (up to 80%). However, the cytochrome pathway respiration partially inhibited (about 50%) by myxothiazol decreases when the Q reduction level increases above 80%. The decrease can be explained by the Q cycle mechanism of complex III. It is also shown that BHAM has an influence on the relationship between the rate of ADP phosphorylation and the Q reduction level when alternative oxidase is active, and that KCN has an influence on the relationship between the alternative oxidase activity and the Q reduction level. These unexpected effects of BHAM and KCN observed at a given Q reduction level are likely due to functional connections between the two pathways activities or to protein-protein interaction. PMID- 11860177 TI - The quinol:fumarate oxidoreductase from the sulphate reducing bacterium Desulfovibrio gigas: spectroscopic and redox studies. AB - The membrane bound fumarate reductase (FRD) from the sulphate-reducer Desulfovibrio gigas was purified from cells grown on a fumarate/sulphate medium and extensively characterized. The FRD is isolated with three subunits of apparent molecular masses of 71, 31, and 22 kDa. The enzyme is capable of both fumarate reduction and succinate oxidation, exhibiting a higher specificity toward fumarate (Km for fumarate is 0.42 and for succinate 2 mM) and a reduction rate 30 times faster than that for oxidation. Studies by Visible and EPR spectroscopies allowed the identification of two B-type haems and the three iron sulplur clusters usually found in FRDs and succinate dehydrogenases: [2Fe 2S]2+/1+ (S1), [4Fe-4S]2+/1+ (S2), and [3Fe-4S]1+/0 (S3). The apparent macroscopic reduction potentials for the metal centers, at pH 7.6, were determined by redox titrations: -45 and -175 mV for the two haems, and +20 and 140 mV for the S3 and SI clusters, respectively. The reduction potentials of the haem groups are pH dependent, supporting the proposal that fumarate reduction is associated with formation of the membrane proton gradient. Furthermore, co reconstitution in liposomes of D. gigas FRD, duroquinone, and D. gigas cytochrome bd shows that this system is capable of coupling succinate oxidation with oxygen reduction to water. PMID- 11860180 TI - Diazoxide and pinacidil uncouple pyruvate-malate-induced mitochondrial respiration. AB - We investigated the effects of K(ATP) channel openers diazoxide and pinacidil on the respiration rate and membrane potential (deltapsi) of rat heart mitochondria, oxidizing pyruvate and malate. Diazoxide and pinacidil (58.8-1348.3 microM) increased the V2 (-ADP) respiration rate accordingly by 13-208% and 30-273% and decreased the deltapsi by 2-17% and 6-55%. These effects were also similar in the respiration medium without K+. Moreover, carboxyatractyloside completely abolished diazoxide- and pinacidil-induced uncoupling, indicating a role for the mitochondrial adenine nucleotide translocase in this process. PMID- 11860179 TI - Opposite effects of Mn(III) and Fe(III) forms of meso-tetrakis(4-N-methyl pyridiniumyl) porphyrins on isolated rat liver mitochondria. AB - The relevance of porphyrins as therapeutic drugs targeted to mitochondria has been widely recognized. In this work, we studied the action of meso-tetrakis porphyrins (TMPyP) on respiring rat liver mitochondria. Mn(III)TMPyP exerted a protective effect against lipid peroxidation induced by Fe(II) or the azo initiator 4,4'-azobis(4-cyanopentanoic acid) (ABCPA), which partition in the hydrophobic phospholipid moiety, and 2,2'-azobis(2-amidinepropane)dihydrochloride (ABAP), which partitions in the aqueous phase. In contrast, Fe(III)TMPyP itself induced an intense lipid peroxidation, accompanied by mitochondrial permeability transition. Both mesoporphyrins studied promoted a release of mitochondrial state 4 respiration, in the concentration range of 1.0-20 microM. Based on the relative effects of Mn(III)TMPyP against ABAP and ABCPA-induced lipid peroxidation, we believe that meso-tetrakis porphyrins must concentrate preferably at membrane water interfaces. PMID- 11860183 TI - Calcium-dependent regulation of interactions of caldesmon with calcium-binding proteins found in growth cones of chick forebrain neurons. AB - 1. This study was undertaken to determine if caldesmon, calmodulin, S100beta, and neurocalcin delta were present in chick forebrain neurons, and if so, to investigate the interactions of these proteins in the presence of different concentrations of calcium. 2. Immunocytochemistry was used to determine the presence and localization of these proteins in cultured forebrain neurons. Western blotting, gel electrophoresis in the presence of different concentrations of calcium, chemical cross-linking, and affinity chromatography were used to investigate the interactions of these proteins with each other. 3. Our data show that caldesmon and three calcium-binding proteins (S100beta, calmodulin, and neurocalcin 3) are localized in growth cones and neurites of chick forebrain neurons in culture. In the presence of different concentration of calcium, these calcium-binding proteins have different affinities to caldesmon and to each other. S100beta binds with greater affinity than calmodulin to caldesmon, and its ability to bind to caldesmon is regulated by neurocalcin delta. 4. These findings suggest a specific calcium-dependent regulatory pathway for modulating actomyosin during growth cone motility. PMID- 11860181 TI - Thyroid status is a key regulator of both flux and efficiency of oxidative phosphorylation in rat hepatocytes. AB - Thyroid status is crucial in energy homeostasis, but despite extensive studies the actual mechanism by which it regulates mitochondrial respiration and ATP synthesis is still unclear. We studied oxidative phosphorylation in both intact liver cells and isolated mitochondria from in vivo models of severe not life threatening hyper- and hypothyroidism. Thyroid status correlated with cellular and mitochondrial oxygen consumption rates as well as with maximal mitochondrial ATP production. Addition of a protonophoric uncoupler, 2,4-dinitrophenol, to hepatocytes did not mimic the cellular energetic change linked to hyperthyroidism. Mitochondrial content of cytochrome oxidase, ATP synthase, phosphate and adenine nucleotide carriers were increased in hyperthyroidism and decreased in hypothyroidism as compared to controls. As a result of these complex changes, the maximal rate of ATP synthesis increased in hyperthyroidism despite a decrease in ATP/O ratio, while in hypothyroidism ATP/O ratio increased but did not compensate for the flux limitation of oxidative phosphorylation. We conclude that energy homeostasis depends on a compromise between rate and efficiency, which is mainly regulated by thyroid hormones. PMID- 11860182 TI - Iron-induced oxidant stress in nonparenchymal liver cells: mitochondrial derangement and fibrosis in acutely iron-dosed gerbils and its prevention by silybin. AB - Hepatic fibrosis due to iron overload is mediated by oxidant stress. The basic mechanisms underlying this process in vivo are still little understood. Acutely iron-dosed gerbils were assayed for lobular accumulation of hepatic lipid peroxidation by-products, oxidant-stress gene response, mitochondrial energy dependent functions, and fibrogenesis. Iron overload in nonparenchymal cells caused an activation of hepatic stellate cells and fibrogenesis. Oxidant-stress gene response and accumulation of malondialdehyde-protein adducts were restricted to iron-filled nonparenchymal cells, sparing nearby hepatocytes. Concomitantly, a significant rise in the mitochondrial desferrioxamine-chelatable iron pool associated with the impairment of mitochondrial oxidative metabolism and the hepatic ATP decrease, was detected. Ultrastructural mitochondrial alterations were observed only in nonparenchymal cells. All biochemical and functional derangements were hindered by in vivo silybin administration which blocked completely fibrogenesis. Iron-induced oxidant stress in nonparenchymal cells appeared to bring about irreversible mitochondrial derangement associated with the onset of hepatic fibrosis. PMID- 11860185 TI - Regulation of corticotropin-releasing hormone (CRH) transcription and CRH mRNA stability by glucocorticoids. AB - 1. The increases in corticotropin-releasing hormone (CRH) mRNA following long term adrenalectomy are associated with low levels of CRH gene transcription, suggesting that glucocorticoids regulate CRH mRNA at the posttranscriptional level. In this study we determined the time course of transcriptional activation after early adrenalectomy by intronic in situ hybridization, and evaluated the effects of glucocorticoids on CRH mRNA stability. 2. Plasma corticosterone was undetectable 3 h after adrenalectomy, but CRH hnRNA increased only by 12 h, and remained elevated for the next 72 h. CRH mRNA increased 18 h after adrenalectomy and reached a plateau lasting from 2 to 6 days, despite very low CRH hnRNA levels. 3. Assessment of CRH mRNA stability, by incubation of slide-mounted hypothalamic sections in an intracellular-like medium at 37 degrees C, prior to measuring CRH mRNA levels by in situ hybridization, revealed a half-life (t1/2) of 11.5 min in sham-operated rats, and a slower decrease adrenalectomized rats (t1/2--26.3 min). Corticosterone administration for 3 days markedly decreased CRH mRNA t1/2 in both sham-operated and adrenalectomized rats (6.5 and 5.0 min, respectively). 4. The data show that adrenalectomy causes transient increases in CRH mRNA transcription, followed by decreases in the rate of CRH mRNA degradation. This suggests that glucocorticoids regulate CRH mRNA at two sites, by inhibiting transcription and by decreasing mRNA stability. PMID- 11860184 TI - Modulation of antagonist binding to serotonin1A receptors from bovine hippocampus by metal ions. AB - 1. The serotonin1A (5-HT1A) receptors are members of a superfamily of seven transmembrane domain receptors that couple to G-proteins. They appear to be involved in various behavioral and cognitive functions. Although specific 5-HT1A agonists have been discovered more than a decade back, the development of selective 5-HT1A antagonists has been achieved only recently. 2. We have examined the modulation of the specific antagonist [3H]p-MPPF binding to 5-HT1A receptors from bovine hippocampal membranes by monovalent and divalent metal ions. Our results show that the antagonist binding to 5-HT1A receptors is inhibited by both monovalent and divalent cations in a concentration-dependent manner. This is accompanied by a concomitant reduction in binding affinity. 3. Our results also show that the specific antagonist p-MPPF binds to all available receptors in the bovine hippocampal membrane irrespective of their state of G-protein coupling and other serotonergic ligands such as 5-HT and OH-DPAT effectively compete with the specific antagonist [3H]p-MPPF. 4. These results are relevant to ongoing analyses of the overall modulation of ligand binding in G-protein-coupled seven transmembrane domain receptors. PMID- 11860186 TI - H9724, a monoclonal antibody to Borrelia burgdorferi's flagellin, binds to heat shock protein 60 (HSP60) within live neuroblastoma cells: a potential role for HSP60 in peptide hormone signaling and in an autoimmune pathogenesis of the neuropathy of Lyme disease. AB - Although Borrelia burgdorferi, the causative agent of Lyme disease, is found at the site of many disease manifestations, local infection may not explain all its features. B. burgdorferi's flagellin cross-reacts with a component of human peripheral nerve axon, previously identified as heat shock protein 60 (HSP60). The cross-reacting epitopes are bound by a monoclonal antibody to B. burgdorferi's flagellin, H9724. Addition of H9724 to neuroblastoma cell cultures blocks in vitro spontaneous and peptide growth-factor-stimulated neuritogenesis. Withdrawal of H9724 allows return to normal growth and differentiation. Using electron microscopy, immunoprecipitation and immunoblotting, and FACS analysis we sought to identify the site of binding of H9724, with the starting hypotheses that the binding was intracellular and not identical to the binding site of II 13, a monoclonal anti-HSP60 antibody. The current studies show that H9724 binds to an intracellular target in cultured cells with negligible, if any, surface binding. We previously showed that sera from patients with neurological manifestations of Lyme disease bound to human axons in a pattern identical to H9724's binding; these same sera also bind to an intracellular neuroblastoma cell target. II-13 binds to a different HSP60 epitope than H9724: II-13 does not modify cellular function in vitro. As predicted, II-13 bound to mitochondria, in a pattern of cellular binding very different from H9724, which bound in a scattered cytoplasmic, nonorganelle-related pattern. H9724's effect is the first evidence that HSP60 may play a role in peptide-hormone-receptor function and demonstrates the modulatory potential of a monoclonal antibody on living cells. PMID- 11860187 TI - Analysis of gene expression following sciatic nerve crush and spinal cord hemisection in the mouse by microarray expression profiling. AB - 1. The responses of periphery (PNS) and central nervous systems (CNS) towards nerve injury are different: while injured mammalian periphery nerons can successfully undergo regeneration, axons in the central nervous system are usually not able to regenerate. 2. In the present study, the genes which were differentially expressed in the PNS and CNS following nerve injury were identified and compared by microarray profiling techniques. 3. Sciatic nerve crush and hemisection of the spinal cord of adult mice were used as the models for nerve injury in PNS and CNS respectivey. 4. It was found that of all the genes examined, 14% (80/588) showed changes in expression following either PNS or CNS injury, and only 3% (18/588) showed changes in both types of injuries. 5. Among all the differentially expressed genes, only 8% (6/80) exhibited similar changes in gene expression (either up- or down-regulation) following injury in both PNS and CNS nerve injuries. 6. Our results indicated that microarray expression profiling is an efficient and useful method to identify genes that are involved in the regeneration process following nerve injuries, and several genes which are differentially expressed in the PNS and/or CNS following nerve injuries were identified in the present study. PMID- 11860189 TI - Correlation of increased grooming behavior and motor activity with alterations in nigrostriatal and mesolimbic catecholamines after alpha-melanotropin and neuropeptide glutamine-isoleucine injection in the rat ventral tegmental area. AB - 1. We wished to further study the behavioral effects of alpha-melanotropin (alpha MSH), melanin-concentrating hormone (MCH), and neuropeptide glutamine-isoleucine (NEI). 2. To this effect we administered alpha-MSH, MCH, and NEI in the ventral tegmental area of the rat, a structure where these neuropeptides are highly concentrated. To further elucidate the biochemical mechanisms of the behavioral effect of these neuropeptides, we determined the degree of grooming behavior and the levels of catecholamines. after neuropeptide administration. 3. We preselected those animals responding to the central injection of alpha-MSH with excessive grooming behavior. We administered the neuropeptides at the dose of 1 microg/0.5 microL, in each side of the ventral tegmental area, bilaterally. We studied grooming behavior, locomotor activity, and total behavior scores, 30 and 65 min after administration of the peptides. 4. Three groups of animals were decapitated immediately after the injection of the neuropeptides, and 30 or 65 min after injection. We measured dopamine (DA), noradrenaline (NA), and the dopac/dopamine ratio (DOPAC/DA) to determine steady state levels of catecholamines and an indirect measure of DA release and metabolism, respectively. 5. Injections of alpha-MSH produced significant elevations in grooming behavior, locomotor activity, and total behavior scores, both 30 and 65 min after peptide administration. This was correlated with significant decreases in DA content, increases in DOPAC content, and increases in the DOPAC/DA ratio. In the caudate putamen, changes in catecholamines occurred both at 30 and 65 min after injection. In the nucleus accumbens, changes were present at 65 min after injection. Conversely, there were no alterations in NA content, either in the caudate putamen or in the nucleus accumbens, at any time after the injection. 6. Injections of NEI resulted in significant elevations in grooming behavior, locomotor activity, and total behavior scores, both 30 and 65 min after peptide administration. This was correlated with increased DOPAC/DA ratio in the nucleus caudatus but not in the nucleus accumbens. Conversely, NEI produced increased NA concentrations in the nucleus accumbens, but not in the nucleus caudatus. 7. Injections of MCH did not produce significant changes in behavior or significant changes in nucleus caudatus or nucleus accumbens catecholamines. 8. Our results indicate (a) There is a correlation with alterations in behavior as induced for the neuropeptides injected here, and changes in extrapyramidal catecholamines. (b) There is a correlation between alterations in behavior and increases in DOPAC/DA ratio in the nucleus caudatus. (c) There is a correlation between alterations in behavior and alterations in catecholamines in the nucleus accumbens. In the nucleus accumbens, DOPAC/DA ratio is changed after alpha-MSH, and NA ratio is changed after NEI injection. (d) Absence of alterations in extrapyramidal catecholamines, and in particular in catecholamines in the nucleus accumbens, correlates with absence of behavioral alterations after neuropeptide administration to the ventral tegmental area. 9. In conclusion, the behavioral effect of exogenous administration of neuropeptides in the ventral tegmental area is peptide-specific, and is probably associated with alterations in catecholamine metabolism and release in the nucleus caudatus and the nucleus accumbens. Both alpha-MSH and NEI seem to stimulate the nigrostriatal DA system. While alpha-MSH appears to stimulate the mesolimbic DA system as well, NEI may exert its actions not through the DA, but through the NA mesolimbic system. The precise contribution of DA and NA, and the relative role of the nucleus caudatus and nucleus accumbens in these behaviors remain to be elucidated. PMID- 11860188 TI - Proteasome activity is critical for the cAMP-induced differentiation of neuroblastoma cells. AB - 1. The ubiquitin-proteasome pathway is involved in a variety of cellular functions in mammalian cells. The role of proteasome, however, in the course of cell differentiation is not well characterized. We hypothesized that proteasome activity might be essential during neuronal cell differentiation. 2. To investigate the role of proteasome during neuronal differentiation, we made use of a murine neuroblastoma cell line (NBP2) that terminally differentiates into mature neurons upon elevation of the intracellular level of adenosine 3',5' cyclic monophosphate (cAMP). To monitor proteasome activity in NBP2 cells, we integrated an expression cassette for a short-lived green fluorescent protein (d2EGFP) into these cells, which were designated as NBP2-PN25. When NBP2-PN25 cells were treated with a proteasome inhibitor, lactacystin or MG132, a dose dependent increase in the constitutive levels of d2EGFP expression was detected. 3. We also found that proteasome inhibition by lactacystin during the cAMP induced differentiation of NBP2-PN25 cells triggered cell death. Both lactacystin and cAMP induction reduced the expression of mRNA for the differentiation associated genes, such as N-myc and cyclin B1. While cAMP-inducing agents decreased the level of N-myc and cyclin B1 proteins, lactacystin increased the level of these proteins. 4. Our data suggest that a reduced level of N-myc and cyclin B1 proteins is critical to commence differentiation, and this can be blocked by a proteasome inhibitor, leading to cell death. Concomitant induction of differentiation and proteasome inhibition, may, therefore, be potentially useful for the treatment of human neuroblastomas. PMID- 11860190 TI - The rate of Tau synthesis is differentially regulated during postnatal development in mouse cerebellum. AB - 1. Tau, which is a microtubule-associated protein, with mRNA targeted to the axon and growth cone, is involved in axonal elongation. During postnatal development in mouse, Tau expression in cerebellar granule cells is reduced afte the second postnatal week. The aim of this work was to study the regulation of the rate of the synthesis of Tau protein during the period of granule cell axonal growth in mouse cerebellum. 2. We found four [35S]methionine-labeled isoforms of Tau synthesized postnataly. Their levels remain constant from postnatal day 9 to 12 (P9-P12), and decreased by P20. 3. The rate of Tau synthesis showed differences with the rate of synthesis of total proteins. They also differ from proteins phosphatases 2A and 2B, both associated with the regulation of Tau function. In addition, the turnover of newly synthesized Tau increased at P20, compared with P9 and P12. 4. These results imply a specific developmental regulation of mRNA translation of Tau, and indicate that, after the period of synapse formation is complete, and therefore axonal growth has finished (P20), only a limited number of new Tau molecules are synthesized. This might reflect that, after synapse formation is complete, newly synthesized Tau molecules are not longer needed. PMID- 11860192 TI - A response to danger. PMID- 11860191 TI - Expression of TrkB receptors in developing visual cortex is not regulated by light. AB - 1. Neurotrophins are very good candidates which relate electrical activity to molecular changes in activity-dependent phenomena. They exert their action through binding to specific tyrosine-kinase receptors: Trk receptors. It is important to consider Trk distribution in order to understand better the role of neurotrophins in the Central Nervous System (CNS). We focused our attention on brain-derived neurotrophic factor (BDNF) Trk receptors (TrkB) during development of the rat visual cortex, since this neurotrophin has been shown to play an important role in visual system development and plasticity. 2. We investigated the full length form of TrkB receptors considering both its total amount and its cellular distribution. To address this issue we used an antibody that recognizes the full length form of TrkB and we used it both in Western blot and immunohistochemistry. 3. We found that the expression of TrkB receptor increases during development, but that there is no effect on visual experience, since dark reared animals show the same protein level and pattern of TrkB expression compared to age-matched, normally reared controls. PMID- 11860193 TI - Poor hand hygiene leads to new guidelines. PMID- 11860194 TI - Access to care: identifying need, mobilizing resources, and finding solutions. PMID- 11860195 TI - State programs face challenges in widening access. PMID- 11860196 TI - Building a new renaissance. PMID- 11860197 TI - An exciting, challenging and frustrating year. PMID- 11860198 TI - Scent of a dental office. PMID- 11860199 TI - Differential esterase expression in developmental mutants of Aspergillus nidulans. AB - Esterase isozymes were used to detect substrate-preference polymorphism in five strains of Aspergillus nidulans, and to show differential gene expression in developmental mutants in response to 5-azacytidine treatment. The medusa mutants B116, SM23, and M25 were selected in the presence of 5-azacytidine (5AC); also the G839 bristle mutant obtained in the absence of 5AC as well as the UT196 master strain and the normal segregant SM24 were used for the esterase studies. The esterase isozyme patterns of the A. nidulans strains observed with 4 methylumbelliferyl esters and alpha- and beta-naphthyl esters indicated a total of 18 isoesterases. Substrate preference for either 4-methylumbelliferyl esters and alpha- or beta-naphthyl esters was observed. Similarity between the different A. nidulans genotypes was 84.4-100%. The genomic similarity of the B116, SM23, and M25 mutant strains (100%) supports previous observations that specific DNA sequences might be targets for 5AC action in this filamentous fungus, and the differential expression of the Est-4 isozyme in the medusa developmental mutant and the Est-2 isozyme specifically detected in the bristle mutant G839 seems to indicate esterase isozymes as possible markers of biochemical differences among different developmental mutants of A. nidulans. PMID- 11860201 TI - Analysis of genetic factors associated with nitric oxide production in mice. AB - Different mouse strains possessing the Nramp1r allele, which were theoretically expected to have relatively high nitric oxide (NO) production after cytokine stimulation, were used to analyze the genetic factors associated with NO production. After gamma interferon and lipopolysaccharide stimulation, the strains NZB/N, DBA/2N, AKR/N, and A/J showed significantly low NO production; NJL, 129/J, MOG, SJL/J, CBA/N, and NOD/Shi had moderate amounts; and C3H/He and SPR had the highest levels as compared to the other mice. The F1 progeny of A/J x C3H/He and AKR/N x C3H/He showed significantly higher NO production, whereas the F1 progeny of DBA/2N x C3H/He produced a relatively low amount. Furthermore, the backcross progeny from their F1 showed variations in NO production, and therefore it was speculated that the regulation of NO production is polygenic. Genetic typing experiments related to the NO production in the backcross progeny demonstrated significant deviations to some genetic microsatellite markers. Sequencing of the iNOS promoter regions of the Nramp1r strains to examine the relationship with NO production revealed that MOG and SPR strains had substitutions within the NF-kappaB and the gamma-IRE transcription binding factor, respectively. PMID- 11860200 TI - Cloning, tissue expression pattern, and chromosome location of a novel human gene BRI3BP. AB - A novel cDNA fragment was identified from a human fetal brain cDNA library by using the coding sequence of human BRI3 gene (Accession No. NM015379) as bait in a yeast two-hybrid screening. Then by 5'-RACE (rapid amplification of cDNA end) and electronic hybridization, we obtained a 1.9 kb contig which consists of a novel gene. It was designated as BRI3BP by the HUGO Nomenclature Committee. It contains an open reading frame encoding 251 amino acids. The calculated molecular weight of the deduced protein is 27.8 kU. The predicted isoelectric point is 9.48. Northern hybridization showed its mRNA was highly expressed in brain, kidney, and liver. By RH mapping, the BRI3BP gene was mapped to human chromosome 12q24.2-qter PMID- 11860202 TI - Phylogenetics of freshwater black basses (Centrarchidae: Micropterus) inferred from restriction endonuclease analysis of mitochondrial DNA. AB - Geographic isolation and habitat specialization has aided in the evolution and genetic integrity of the micropterid bass species of North America. Members of the genus Micropterus form a close natural unit with little morphologic and meristic variation. Our goals were to measure the genetic characteristics of and distances between six black bass species by using mitochondrial DNA analysis. Mitochondrial DNA restriction fragment length polymorphisms were examined in Guadalupe bass (M. treculi), largemouth bass (M. salmoides), shoal bass (M. cataractae), smallmouth bass (M. dolomieu), spotted bass (M. punctulatus), and Suwannee bass (M. notius), using 15 restriction endonucleases. The bluegill (Lepomis macrochirus) was used as an outgroup. The phylogeny inferred from Dollo parsimony cladistic analysis concurred with published results from allozyme analyses, yet it was inconsistent with published meristic analyses. Genetic distances between species ranged from 0.0659 to 0.2145, with the largemouth and Suwannee basses showing the greatest divergence from the other black basses. The Guadalupe, smallmouth, and spotted basses were most diverged from the bluegill. The black basses diverged over a broad time frame, with estimated black bass speciation occurring during late Miocene-early Pliocene (3.30-10.73 MYA). PMID- 11860203 TI - Specific frequency distribution of erythrocytic X-protein alleles in indigenous sheep populations in East Asia. AB - The nonhemoglobin erythrocytic X-protein polymorphism consisting of two phenotypes called X-positive [X(+)] and X-negative [X(-)] was determined in 576 unrelated healthy native sheep of East Asia, using one-dimensional and horizontal starch gel electrophoresis. A striking difference in the frequency of the X allele coding dominantly for the X(+) type between the northern and southern populations of native East Asian sheep divided by the boundary of the Himalaya Mountains was seen (P < 0.0001). The X allele frequency ranged from 0 to 0.0438 with an average of 0.0323 in the northern population examined, consisting of the Bhyanglung, Baruwal, Yunnan, and Khalkhas sheep belonging to the Tibetan and Mongolian sheep groups. In contrast, the frequency of the same allele was in the range of 0.2037-0.4655 and the mean frequency was 0.2998 in the southern population tested, consisting of the Bengal, Kagi, Lampuchhre, Vietnamese, and Myanmar sheep, which belong to the Indian sheep group. This finding suggests that the X allele appears to be an Indian sheep marker and is potentially important in phylogenetic studies on native sheep populations, especially in East Asia. PMID- 11860204 TI - Mitochondrial cytochrome b gene sequence diversity in the Korean hare, Lepus coreanus Thomas (Mammalia, Lagomorpha). AB - Partial sequences of the mitochondrial cytochrome b gene of the Korean hare (Lepus coreanus) were analyzed to determine the degree of genetic diversity. Nine haplotypes were observed, and the maximum Tamura-Nei nucleotide distance among them was 2.8%, indicating that genetic diversity of L. coreanus is moderate. In order to clarify the Korean hare's taxonomic status and relationship with the Manchurian hare (L. mandshuricus) and the Chinese hare (L. sinensis), these nine haplotypes of the Korean hare were compared with 13 haplotypes from five other species of eastern Asian Lepus including L. mandshuricus and L. sinensis. The Korean hare was distinct in its cytochrome b gene, and it is confirmed that L. coreanus is a valid species, as noted by Jones and Johnson (1965, Univ. Kansas Publ. (Mus. Nat. Hist.) 16:357). Further analyses of mtDNA cytochrome b gene with additional specimens of L. coreanus from North Korea and other species of Lepus from eastern Asia are needed to clarify the taxonomic status of the divergent mtDNA clades of L. mandshuricus and L. sinensis. PMID- 11860205 TI - Microarray data quality analysis: lessons from the AFGC project. Arabidopsis Functional Genomics Consortium. AB - Genome-wide expression profiling with DNA microarrays has and will provide a great deal of data to the plant scientific community. However, reliability concerns have required the development data quality tests for common systematic biases. Fortunately, most large-scale systematic biases are detectable and some are correctable by normalization. Technical replication experiments and statistical surveys indicate that these biases vary widely in severity and appearance. As a result, no single normalization or correction method currently available is able to address all the issues. However, careful sequence selection, array design, experimental design and experimental annotation can substantially improve the quality and biological of microarray data. In this review, we discuss these issues with reference to examples from the Arabidopsis Functional Genomics Consortium (AFGC) microarray project. PMID- 11860206 TI - Large-scale plant proteomics. AB - Large-scale and high throughput approaches increasingly play an essential role in the study of biological systems, which are per se highly complex. Therefore, they need to be examined by these extensive methods to receive information about the large genomic and proteomic networks. In plant biology, this purpose has a strong support through the accessability of the complete genome sequence of the model plant Arabidopsis thaliana. This brief review intends to focus on the basics and the state-of-the-art of these high-throughput technologies and their application to plant proteomics. It describes protein microarrays, the use of antibodies, 2 DE and MS methods and the yeast two hybrid system, which are emerging as the major technologies for plant proteomics. PMID- 11860208 TI - Gene replacement by homologous recombination in plants. AB - After the elucidation of the sequence of the yeast genome a major effort was started to elucidate the biological function of all open reading frames of this organisms by targeted gene replacement via homologous recombination. The establishment of the complete sequence of the genome of Arabidopsis thaliana would principally allow a similar approach. However, over the past dozen years all attempts to establish an efficient gene targeting technique in flowering plants were in the end not successful. In contrast, in Physcomitrella patens an efficient gene targeting procedure has been set up, making the moss a valuable model system for plant molecular biologists. But also for flowering plants recently several new approaches--some of them based on the availability of the genomic sequence of Arabidopsis--were initiated that might finally result on the set up of a general applicable technique. Beside the production of hyper recombinogenic plants either via expression or suppression of specific gene functions or via undirected mutagenesis, the application of chimeric oligonucleotides might result in major progress. PMID- 11860207 TI - Metabolomics--the link between genotypes and phenotypes. AB - Metabolites are the end products of cellular regulatory processes, and their levels can be regarded as the ultimate response of biological systems to genetic or environmental changes. In parallel to the terms 'transcriptome' and proteome', the set of metabolites synthesized by a biological system constitute its 'metabolome'. Yet, unlike other functional genomics approaches, the unbiased simultaneous identification and quantification of plant metabolomes has been largely neglected. Until recently, most analyses were restricted to profiling selected classes of compounds, or to fingerprinting metabolic changes without sufficient analytical resolution to determine metabolite levels and identities individually. As a prerequisite for metabolomic analysis, careful consideration of the methods employed for tissue extraction, sample preparation, data acquisition, and data mining must be taken. In this review, the differences among metabolite target analysis, metabolite profiling, and metabolic fingerprinting are clarified, and terms are defined. Current approaches are examined, and potential applications are summarized with a special emphasis on data mining and mathematical modelling of metabolism. PMID- 11860209 TI - Recombinase-directed plant transformation for the post-genomic era. AB - Plant genomics promises to accelerate genetic discoveries for plant improvements. Machine-driven technologies are ushering in gene structural and expressional data at an unprecedented rate. Potential bottlenecks in this crop improvement process are steps involving plant transformation. With few exceptions, genetic transformation is an obligatory final step by which useful traits are engineered into plants. In addition, transgenesis is most often needed to confirm gene function, after deductions made through comparative genomics, expression profiles, and mutation analysis. This article reviews the use of recombinase systems to deliver DNA more efficiently into the plant genome. PMID- 11860211 TI - Computational gene finding in plants. AB - Automated methods for identifying protein coding regions in genomic DNA have progressed significantly in recent years, but there is still a strong need for more accurate computational solutions to the gene finding problem. Large-scale genome sequencing projects depend greatly on gene finding to generate accurate and complete gene annotation. Improvements in gene finding software are being driven by the development of better computational algorithms, a better understanding of the cell's mechanisms for transcription and translation, and the enormous increases in genomic sequence data. This paper reviews some of the most widely used algorithms for gene finding in plants, including technical descriptions of how they work and recent measurements of their success on the genomes of Arabidopsis thaliana and rice. PMID- 11860210 TI - Plant genome evolution: lessons from comparative genomics at the DNA level. AB - Angiosperm genomes show tremendous variability in genome size and chromosome number. Nevertheless, comparative genetic mapping has revealed genome collinearity of closely related species. Sequence-based comparisons were used to assess the conservation of gene arrangements. Numerous small rearrangements, insertions/deletions, duplications, inversions and translocations have been detected. Importantly, comparative sequence analyses have unambiguously shown micro-collinearity of distantly related plant species. Duplications and subsequent gene loss have been identified as a particular important factor in the evolution of plant genomes. PMID- 11860212 TI - Computational modeling of gene structure in Arabidopsis thaliana. AB - Computational gene identification by sequence inspection remains a challenging problem. For a typical Arabidopsis thaliana gene with five exons, at least one of the exons is expected to have at least one of its borders predicted incorrectly by ab initio gene finding programs. More detailed analysis for individual genomic loci can often resolve the uncertainty on the basis of EST evidence or similarity to potential protein homologues. Such methods are part of the routine annotation process. However, because the EST and protein databases are constantly growing, in many cases original annotation must be re-evaluated, extended, and corrected on the basis of the latest evidence. The Arabidopsis Genome Initiative is undertaking this task on the whole-genome scale via its participating genome centers. The current Arabidopsis genome annotation provides an excellent starting point for assessing the protein repertoire of a flowering plant. More accurate whole-genome annotation will require the combination of high-throughput and individual gene experimental approaches and computational methods. The purpose of this article is to discuss tools available to an individual researcher to evaluate gene structure prediction for a particular locus. PMID- 11860214 TI - Surviving in a sea of data: a survey of plant genome data resources and issues in building data management systems. AB - Exponential growth of data, largely from whole-genome analyses, has changed the way biologists think about and handle data. Optimal use of these data requires effective methods to analyze and manage these data sets. Computers, software and the World Wide Web are now integral components of biological discovery. Understanding how information is obtained, processed and annotated in public databases allows researchers to effectively organize, analyze and export their own data into these databases. In this review we focus largely on two areas related to management of genomic data. We cite examples of resources available in the public domain and describe some of the software for data management systems currently available for plant research. In addition, we discuss a few concepts of data management from the perspective of an individual or group that wishes to provide data to the public databases, to use the information in the public databases more efficiently, or to develop a database to manage large data sets internally or for public access. These concepts include data descriptions, exchange format, curation, attribution, and database implementation. PMID- 11860213 TI - Through a genome, darkly: comparative analysis of plant chromosomal DNA. AB - Plant nuclear genomes encompass a wide range of variation in size and nucleotide composition with diverse arrangements of chromosomal segments, repetitive sequences and distribution of genes. Comparative genomic analysis may be undertaken at different levels of organisation, which are reflected in this review, together with a focus on the genetic and functional significance of the observed variation. Patterns of genome organisation have been revealed which reflect the different underlying mechanisms and constraints driving change. Thus comparative issues of genome size, nucleotide sequence composition and genome heterogeneity are provided as a background to understanding the different levels of segmental and repetitive sequence duplication and distribution of genes. The extent of synteny and collinearity revealed by recent genetic and sequence comparisons is discussed, together with a consideration of problems associated with such analyses. The possible origins and mechanisms of variation in genome size and organisation are covered, including the prevalence of duplication at different levels of organisation. The likely genetic, functional and adaptive consequences of replicated loci are discussed with evidence from comparative studies. The scope for comparative analysis of epigenetic plant genome variation is considered. Finally, opportunities for applying comparative genomics to isolating genes and understanding complex crop genomes are addressed. PMID- 11860217 TI - Grapefruit juice effects on the bioavailability of cyclosporin-A in rats. AB - Previous studies indicate that blood levels of cyclosporin-A are increased by concomittant administration of grapefruit juice in healthy subjects and patients. It was suggested that grapefruit juice could inhibit the metabolism of cyclosporin-A by CYP3A4, the predominant cytochrome P450 enzyme in the gut wall and liver. However, up to date, the mechanism of action of grapefruit juice has not been conclusively identified and no work has been conducted in animals to quantify its effect on cyclosporin-A metabolism. This study compared the disposition of cyclosporin-A (5 mg/kg) coadministered with grapefruit juice, orange juice or water (10 ml/kg) in male Sprague-Dawley rats. Time to peak concentration was about 5 h for each group. Area under the blood concentration time curve and peak concentration of cyclosporin-A were increased by 31% and 20%, respectively, with grapefruit juice (P < 0.05). The effects of grapefruit juice were not duplicated by orange juice which did not differ significantly from water for any of the parameters tested. These results confirm that grapefruit juice may act as an inhibitor of drug metabolism altering the disposition of concomittantly administered cyclosporin-A in rats. Nonetheless, it was demonstrated that, under appropriate experimental conditions, rats may be suitable models for in vivo investigation of the interaction mechanism between grapefruit juice and cyclosporin-A. PMID- 11860218 TI - Beclomethasone dipropionate versus budesonide inhalation suspension in children with mild to moderate persistent asthma. AB - Inhaled steroids are the most effective long-term treatment of persistent asthma but many children are unable to use correctly the available inhalers. Administration of nebulized corticosteroids has some advantages over the administration with pressurised metered-dose inhalers (pMDls). The objective of this multicenter randomised study was to compare the efficacy and tolerability of nebulized corticosteroids in paediatric patients with asthma. 127 patients aged > or = 6 and < or = 14 years with a diagnosis of mild to moderate persistent asthma (PEFR % predicted > 50% and < 85%) and positive response to the reversibility test were randomized. The patients were assigned by randomisation to one of the two treatment groups (4 weeks): beclomethasone dipropionate (BDP) 800 microg/daily b.i.d. (n = 66) or budesonide (BUD) 1000 microg/daily b.i.d. (n = 61) both administered by nebulizer. The primary efficacy end point was the final mean of PEFR measured at clinical visit (clinic PEFR). In the BDP group clinic PEFR increased from 177.5 +/- 80 L/min to 246.6 +/- 84.2 L/min (p < 0.001 vs baseline), while in the BUD group the increase was from 180.4 +/- 77.8/min to 260.9 +/- 84.1 L/min (p < 0.001 vs baseline) (NS between treatments). FEV1 (% predicted) increased from 77.8% to 92.7% (p < 0.001 vs baseline) and from 74.1% to 95.9% (p < 0.001 vs baseline) in BDP and BUD group respectively (NS between treatments). Patients reduced the use of salbutamol rescue medication by 76% and 81% in BDP and BUD group respectively (p < 0.001 vs baseline, NS between treatments). 4 patients in the BDP group and 2 in the BUD group reported adverse events (AEs). AEs were mild to moderate and never there was the need to discontinue the treatments. In conclusion the results of this study demonstrate that both BDP (800 microg/daily) and BUD (1000 microg/daily) administered by nebulization are effective and with a acceptable safety and tolerability profile. PMID- 11860216 TI - DNA microarrays for functional plant genomics. AB - DNA microarray technology is a key element in today's functional genomics toolbox. The power of the method lies in miniaturization, automation and parallelism permitting large-scale and genome-wide acquisition of quantitative biological information from multiple samples. DNA microarrays are currently fabricated and assayed by two main approaches involving either in situ synthesis of oligonucleotides ('oligonucleotide microarrays') or deposition of pre synthesized DNA fragments ('cDNA microarrays') on solid surfaces. To date, the main applications of microarrays are in comprehensive, simultaneous gene expression monitoring and in DNA variation analyses for the identification and genotyping of mutations and polymorphisms. Already at a relatively early stage of its application in plant science, microarrays are being utilized to examine a range of biological issues including the circadian clock, plant defence, environmental stress responses, fruit ripening, phytochrome A signalling, seed development and nitrate assimilation. Novel insights are obtained into the molecular mechanisms co-ordinating metabolic pathways, regulatory and signalling networks. Exciting new information will be gained in the years to come not only from genome-wide expression analyses on a few model plant species, but also from extensive studies of less thoroughly studied species on a more limited scale. The value of microarray technology to our understanding of living processes will depend both on the amount of data to be generated and on its clever exploration and integration with other biological knowledge arising from complementary functional genomics tools for 'profiling' the genome, proteome, metabolome and phenome. PMID- 11860215 TI - Comprehensive gene expression analysis by transcript profiling. AB - After the completion of the genomic sequence of Arabidopsis thaliana, it is now a priority to identify all the genes, their patterns of expression and functions. Transcript profiling is playing a substantial role in annotating and determining gene functions, having advanced from one-gene-at-a-time methods to technologies that provide a holistic view of the genome. In this review, comprehensive transcript profiling methodologies are described, including two that are used extensively by the authors, cDNA-AFLP and cDNA microarraying. Both these technologies illustrate the requirement to integrate molecular biology, automation, LIMS and data analysis. With so much uncharted territory in the Arabidopsis genome, and the desire to tackle complex biological traits, such integrated systems will provide a rich source of data for the correlative, functional annotation of genes. PMID- 11860219 TI - Genetically modified foods and children potential health risks. AB - AIM: Professor Pusztai was publicly humiliated over claims that genetically modified (GM) Frankenstein food may be harmful. He was stripped of his post and described as 'muddled' by his superiors after he referred to experiments in which rats had been damaged when fed genetically-altered potatoes. Who is in an unsound scenario, supported by verbal expressions ("substantially"), should even more expend further effort in conducting scientific investigation into the safety of GM varieties of plants. OBSERVATIONS: Of particular concern is the exposure of infants and children to GM foods (GMFs) because of their possible increased susceptibility for untoward effects. Several examples stress that the ascertainment of human disease emerged after certain materials were widely used. Studies show that some compounds were not adequately tested for toxicity before their commercial introduction, whereas proper premarked testing would have prevented a prolonged exposure. CONCLUSIONS: Too often the toxicity of these substances is untested and the potential hazards that they may pose to children have not been examined. Nobody has evaluated whether intrauterine and infant exposure to GMFs may have profound permanent and irreversible consequences even in adult life. In this paper we analyse issues pertaining to children's health that have been largely ignored. PMID- 11860220 TI - Eudragit FS 30 D: a new pH-sensitive polymer covering for mesalamine. PMID- 11860222 TI - Inhaled corticosteroid delivery systems: clinical role of a breath-actuated device. AB - Several devices have been developed to overcome the need to co-ordinate actuation with inhalation required during use of a pressurised metered dose inhaler (MDI) and to improve drug delivery to the lung. These include spacer attachments for MDIs, dry powder inhalers and breath-actuated MDIs. The breath-actuated Autohaler (3M Pharmaceuticals) is a compact, multidose inhaler device that, unlike dry powder inhalers, does not rely on the patient's inspiratory effort to aerosolise the dose of medication. Due to its simple operation, the Autohaler is suitable for patients unable to operate a conventional MDI efficiently, including the elderly, children, patients with arthritis and patients with low inspiratory flow rates. The mandatory replacement of chlorofluorocarbon propellants with non-ozone depleting propellants has given the opportunity to improve drug delivery characteristics of MDIs. Recently, a formulation of beclomethasone dipropionate in hydrofluoroalkane-134a (HFA-BDP), has been developed in a conventional MDI that delivers most of the emitted dose to the lung. Drug deposition studies show that the HFA-BDP formulation in the Autohaler device has a similar lung deposition pattern to drug delivered from the MDI, when used correctly, and dose delivery is consistent across a wide range of inspiratory flow rates. Furthermore, HFA-BDP Autohaler has similar clinical benefits to CFC-BDP Autohaler but at less than half the dose. HFA-BDP Autohaler offers a useful CFC-free delivery option for patients challenged by the conventional MDI device. PMID- 11860221 TI - Microalbuminuria as a marker of cardiac damage in essential hypertension. AB - A subclinical elevation in urinary albumin excretion (UAE) microalbuminuria is frequently seen in essential hypertension. The level of blood pressure appears to be an important factor in the development of microalbuminuria. Moreover there is some evidence to indicate that microalbuminuria may be an early marker of increased cardiovascular risk. Aim of this study was to evaluate the prevalence of UAE in hypertensives with normal left ventricular mass and to study any association with blood pressure level and with possible modification in left ventricular function. A group of 112 subjects diagnosed as having stage 1-2 essential hypertension were included in the study. Patients underwent urinary collection to evaluate UAE and to 24/hours arterial blood pressure monitoring. Moreover a complete echocardiography was performed. According with UAE levels patients were divided into three groups: A: UAE 0-15 mg/24 h, B: UAE 16-29 mg/24 h, C: UAE 30-300 mg/24 h. We found a significant correlation between 24/h SBP, 24/h DBP and UAE. We observed a significant correlation between impaired diastolic function and UAE. UAE is influenced by BP levels with better correlation with 24/h systolic values. UAE is associated with subclinical decrease of left ventricular function and may be an early marker of cardiac involvement. PMID- 11860223 TI - Prevention by delay: nonspecific immunity elicited by IL-12 hinders Her-2/neu mammary carcinogenesis in transgenic mice. AB - As a natural consequence of the expression of the activated transforming rat Her 2/neu oncogene all mammary glands of female transgenic BALB/c (BALB-neuT) mice develop atypical epithelial hyperplasia which progresses to invasive carcinoma. A lobular carcinoma is palpable in all mammary glands of 33-week-old BALB-neuT mice. This progression is markedly delayed by systemic administration of IL-12. In a series of studies the best administration schedule, the lowest dose and the most effective administration time have been defined. The cellular and molecular mechanisms resulting in the delay of carcinogenesis have been established. By means of a series of downstream mediators IL-12 inhibits the angiogenic burst that goes along with the passage from preneoplastic to neoplastic and invasive lesions; it also recruits lymphoid cells in the mammary pad and activates their cytotoxicity towards neoplastic cells and newly formed vessels; and furthermore, it induces lymphoid cells to trigger antiangiogenic activities in neoplastic epithelial cells. Effective, low-dose and non-toxic IL-12 treatments may thus be envisaged as a possible option in the management of preneoplastic mammary lesions and in mammary cancer prevention. PMID- 11860224 TI - Simultaneous determination of apoptosis and surface antigen expression in tumor adherent cells. AB - Apoptosis is a physiological, gene-directed form of cell death aimed at controlling cell proliferation in several biological conditions. It plays a crucial role in modulating tissue growth during embryonic development, cell turnover in adult life, and it seems to be the most frequent mechanism of tumor cell deletion by chemotherapy. Flow cytometry is a widely-used technique for checking apoptosis, permitting a multiparametric analysis. It is possible to follow the alterations occurring in the nucleus, mitochondria and plasmatic membrane during the different apoptotic stages using probes such as LDS-751, JC-1 or Annexin V. The potential of these probes to identify the early or late stages of apoptosis has been widely investigated in cells growing in suspension. In order to assess apoptosis in adherent cells, we tested a combination of fluorescein diacetate (FDA), a substrate for non specific esterase whose activity decreases during the early phase of apoptosis, and trypan blue in MCF-7 human breast cancer cells. Apoptotic cells showed a decrease in the green fluorescence emitted by fluorescein, the product of FDA hydrolysis, whereas necrotic cells emitted a red fluorescence due to the trypan blue staining. FDA-trypan blue double-staining was used to investigate the different kinetics of apoptosis induced by taxol, camptothecin and UV-B irradiation in MCF-7 cells. This method is rapid and simple, and can be used for monitoring the process of apoptosis from early stages in adherent cells, for the physical separation of apoptotic and live cells, and for immunophenotyping, including Fas expression. PMID- 11860226 TI - Abrogation of surgery-induced IL-6 hypersecretion by presurgical immunotherapy with IL-2 and its importance in the prevention of postoperative complications. AB - Because of its immunosuppressive effect, surgery-induced immunosuppression may depend at least in part on the postoperative hypersecretion of IL-6, which is also responsible for surgical complications. Most of the immunosuppressive events induced by surgery, including lymphocytopenia, NK and T lymphocyte decline, and dendritic cell deficiency have been proven to be abrogated by a preoperative injection of IL-2 for few days. However, the cytokine mechanisms responsible for IL-2-induced abrogation of surgery-related immunosuppression need to be better investigated and understood. This study was performed to analyze the influence of IL-2 presurgical immunotherapy on IL-6 secretion in the postoperative period. The study was performed in 12 operable colorectal cancer patients, who were preoperatively pretreated with IL-2 (12 million lU/day subcutaneously for 3 consecutive days before surgery). The control group consisted of 21 age-and disease-matched colorectal cancer patients who underwent surgery without a preoperative immunotherapy with IL-2. Serum levels of IL-6 were measured by an enzyme immunoassay before surgery, and at days 3 and 7 of the postoperative period. A significant increase in mean serum levels of IL-6 occurred in the postoperative period only in the control patients, whereas in the IL-2 pretreated group no significant difference was seen between presurgical and postoperative IL 6 mean concentrations. The difference between controls and IL-2 group was particularly evident for patients with abnormally elevated presurgical values of IL-6. This study, by showing that a presurgical injection of IL-2 may prevent surgery-induced IL-6 enhanced secretion, would suggest that the previously described neutralization of surgery-induced immunosuppression by IL-2 preoperative immunotherapy may depend at least in part on the inhibition of postoperative production of IL-6, whose immunosuppressive effects have been well demonstrated at least on anticancer immunity. PMID- 11860225 TI - Decrease of erythropoietin level by human recombinant tumour necrosis factor alpha (hrec TNFalpha) in patients with advanced cancer. AB - Anaemia is a frequent complication of chronic inflammation, infectious diseases and cancer. Inappropriate erythropoietin production is regarded as one of the main causative factors responsible for the occurrence of anaemia. The pathogenesis of TNFalpha induced-anaemia has not been fully clarified yet and its influence on hematopoiesis has been suggested. We performed a clinical study to access the influence of hrec TNFalpha administration on plasma EPO concentration and the degree of anaemia in patients with advanced solid tumours for whom no other kind of therapy but palliative treatment was available. All these patients exposed mild anaemia (HT 36.1 +/- 1.0%). Plasma EPO was estimated at 8 a.m. before and after 5 days of TNFalpha therapy with a dose of 75 pg/day iv (cycle I). Two weeks later plasma EPO was estimated again before and after 5 days of TNFalpha administration of a double dose (150 microg/day) (cycle II). The control group comprised 8 non-cancer patients (5M/3F, age 48.5 +/- 6yr) with the same degree of anaemia (HT 36 +/- 1.1%) due to haemorrhage. In the control group the plasma EPO level was significantly higher (54.2 +/- 8 mU/ml) than in cancer patients before cycle I (17.1 +/- 2.5 mU/ml) and II (14.6 +/- 3.8 mU/ml) respectively.TNF administration was followed by a significant decline of plasma EPO both after the first (17.1 +/- 2.5 vs 9.0 +/- 1.5 mU/ml) and second cycle (14.6 +/- 3.8 vs 8.4 +/- 2.0 mU/ml) of TNF treatment. CONCLUSIONS: Patients with solid cancer and mild anaemia are characterised by inappropriate low plasma EPO concentration. Therapy with TNFalpha exerts a suppressive effect on EPO secretion in these patients. PMID- 11860228 TI - Transdermal transport by hydroelectrophoresis: a novel method for delivering molecules. PMID- 11860227 TI - Altered expression of integrins and basement membrane proteins in malignant and pre-malignant lesions of oral mucosa. AB - Integrins are transmembrane receptors that regulate cell-cell and cell extracellular matrix (ECM) contact. In epithelial tissues, they interact with ECM components of the basement membrane (BM) to maintain the homeostasis and the architecture of the tissue. This interaction controls several cell functions such as adhesion, migration, proliferation, differentiation, and therefore has a key role in cancer development and metastasis. We studied the expression of integrins and ECM components of the BM by immunohistochemistry in frozen specimens of malignant squamous cell carcinoma (SCC), pre-malignant lesions of the oral mucosa (leucoplakia) and oral lichen planus. In invasive SCC, we observed altered polarity and distribution of alpha2beta1, alpha6beta4 and alpha3beta1 integrins, whereas in the in situ carcinoma alpha6beta4 and alpha3beta1 patterns only were altered. Immunostaining for ECM components such as Laminin-1 (Ln-1), Ln-5, and Collagen IV (Coll IV) was discontinuous and interrupted in invasive SCC, whereas it was normal in the in situ carcinoma. In both pre-malignant lesions and lichen planus specimens, integrins were expressed in a polarized manner in the presence of a normal BM, whereas were abnormally distributed in those tissues with altered staining patterns of the ECM components. In conclusion, we suggest that abnormal re-distribution of alpha3beta1 and alpha6beta4 integrins and expression of ECM components such as Ln-5 could play an important role in SCC invasion and metastasis. PMID- 11860229 TI - Human leucocyte differentiation antigens and the human genome project. PMID- 11860230 TI - CD52 (CAMPATH1). PMID- 11860231 TI - CD90. PMID- 11860232 TI - Review finds U.S. animal disease control system adequate, but needing improvement. PMID- 11860233 TI - Global coalition unites to aid animals of Afghanistan zoo. PMID- 11860234 TI - A little bird told me, you'd better listen to the law. PMID- 11860235 TI - Is your state emergency management agency doing all it can to get you the help you need in a disaster? PMID- 11860236 TI - Thinks oral interview should be replaced. PMID- 11860237 TI - Addressing the moral anguish of euthanasia. PMID- 11860238 TI - What is your diagnosis? A sharply marginated soft-tissue mass occluding the lumen of the cervical region of the trachea with possible involvement of the esophagus. PMID- 11860239 TI - Detection of vesicular exanthema of swine-like calicivirus in tissues from a naturally infected spontaneously aborted bovine fetus. PMID- 11860240 TI - Prevalence of serum thyroid hormone autoantibodies in dogs with clinical signs of hypothyroidism. AB - OBJECTIVE: To determine prevalence of thyroid hormone autoantibodies (THAA) in serum of dogs with clinical signs of hypothyroidism. DESIGN: Cohort study. SAMPLE POPULATION: 287,948 serum samples from dogs with clinical signs consistent with hypothyroidism. PROCEDURE: Serum THAA were detected by use of a radiometric assay. Correlation and chi2 analyses were used to determine whether prevalence varied with breed, age, sex, or body weight. Only breeds for which > or = 50 samples had been submitted were used for analysis of breed prevalence. RESULTS: Thyroid hormone autoantibodies were detected in 18,135 (6.3%) samples. The 10 breeds with the highest prevalence of THAA were the Pointer, English Setter, English Pointer, Skye Terrier, German Wirehaired Pointer, Old English Sheepdog, Boxer, Maltese, Kuvasz, and Petit Basset Griffon Vendeen. Prevalence was significantly correlated with body weight and was highest in dogs between 2 and 4 years old. Females were significantly more likely to have THAA than were males. CONCLUSIONS AND CLINICAL RELEVANCE: Thyroid hormone autoantibodies may falsely increase measured triiodothyronine (T3) and thyroxine (T4) concentrations in dogs; results suggest that T3 concentration may be falsely increased in approximately 57 of 1,000 dogs with hypothyroidism and that T4 concentration may be falsely increased in approximately 17 of 1,000 dogs with hypothyroidism. Results also suggested that dogs of certain breeds were significantly more or less likely to have THAA than were dogs in general. PMID- 11860241 TI - Evaluation of a radiographic caudolateral curvilinear osteophyte on the femoral neck and its relationship to degenerative joint disease and distraction index in dogs. AB - OBJECTIVE: To determine prevalence of a radiographic caudolateral curvilinear osteophyte (CCO) on the femoral neck in various breeds and age groups of dogs and to evaluate its contemporaneous relationship with degenerative joint disease (DJD) and distraction index (DI). DESIGN: Cross-sectional prevalence study. ANIMALS: 25,968 dogs, including 3,729 German Shepherd Dogs, 4,545 Golden Retrievers, 6,277 Labrador Retrievers, and 1,191 Rottweilers. PROCEDURE: Data from the University of Pennsylvania Hip Improvement Program database were analyzed, including ventrodorsal hip-extended, compression, and distraction radiographs. The CCO and radiographic signs of DJD were considered independent events and were interpreted as either present or absent. Statistical methods were used to evaluate the CCO as a possible risk factor for DJD and assess its association with DI, as measured by use of distraction radiography. RESULTS: When all breeds were pooled, DJD was detected in 8.6% of dogs, and the CCO was detected in 21.6% of dogs. Among dogs with a CCO, 25.1% had radiographic evidence of DJD. Among dogs without a CCO, only 4% had DJD. Dogs with a CCO were 7.9 times as likely to have DJD as were those without a CCO. Additionally, DI, weight, and age were significant risk factors for the CCO. CONCLUSION AND CLINICAL RELEVANCE: Results confirm the contemporaneous association between the CCO and DJD and that passive hip laxity, as measured by use of the DI, is associated with both the CCO and DJD. PMID- 11860242 TI - Comparison of platelet count recovery with use of vincristine and prednisone or prednisone alone for treatment for severe immune-mediated thrombocytopenia in dogs. AB - OBJECTIVE: To evaluate the effect of prednisone alone, compared with a combination of prednisone and vincristine, on platelet counts in bleeding dogs with severe primary immune-mediated thrombocytopenia (IMT). DESIGN: Prospective case study. ANIMALS: 24 dogs with severe primary IMT PROCEDURE: All dogs received immunosuppressive doses of prednisone (1.5 to 2 mg/kg [0.7 to 0.9 mg/lb] of body weight, PO, q 12 h). In addition, 12 dogs received a single dose of vincristine (0.02 mg/kg [0.01 mg/lb], IV). Platelet count, transfusion requirement, and outcome were monitored. A response was defined as an increase in platelet count to > or = 40,000/microl. Dogs in the prednisone group that failed to respond received 1 dose of vincristine on day 7. RESULTS: Dogs that received prednisone and vincristine had a significantly faster increase in platelet count to > or = 40,000 platelets/microl than dogs that received prednisone alone (mean +/- SD, 4.9 +/- 1.1 vs 6.8 +/- 4.5 days, respectively). A similarly rapid response was observed in dogs that received vincristine on day 7 after treatment with prednisone alone failed. Furthermore, duration of hospitalization was reduced in the vincristine group, compared with the prednisone group (5.4 +/- 0.3 vs 7.3 +/- 0.5 days, respectively). No adverse effects attributable to vincristine were observed in any dog. CONCLUSIONS AND CLINICAL RELEVANCE: Administration of combined vincristine and prednisone is associated with more rapid increase in platelet numbers and shortened duration of hospitalization in dogs with IMT, compared with use of prednisone alone. Early use of vincristine seems warranted in dogs with severe primary IMT. PMID- 11860243 TI - Comparison of hematologic and biochemical values for blood samples obtained via jugular venipuncture and via vascular access ports in cats. AB - OBJECTIVE: To determine whether hematologic and serum biochemical values for blood samples obtained from cats via vascular access ports (VAP) are comparable to those for samples obtained by direct venipuncture. DESIGN: Prospective study. ANIMALS: 14 healthy cats. PROCEDURE: A VAP was surgically implanted in a jugular vein in each cat. Blood samples were obtained from the VAP and by direct venipuncture of the contralateral jugular vein 10 weeks after VAP placement. Results of hematologic and serum biochemical analyses were compared by use of a paired t-test. The Pvalue to reject the null hypothesis was adjusted to account for multiple comparisons by using the Bonferroni procedure in which the nominal P to-reject value is divided by the number of comparisons (0.05/24 = 0.002). RESULTS: Paired samples (VAP and venipuncture) obtained 10 weeks after VAP placement were evaluated for each cat. Of the 24 measured analytes, only potassium, total protein, and albumin concentrations differed significantly (P< 0.001 for all 3) between VAP and venipuncture samples. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that samples obtained from VAP are suitable for routine hematologic monitoring of feline cancer patients. Sample hemolysis may account for a slight increase in potassium, total protein, and albumin concentrations obtained from VAP samples. However, the values of variables most critical for monitoring of patients receiving chemotherapy (ie, mature neutrophil and platelet counts) are comparable. If proper techniques are used, VAP may be used for administration of chemotherapy as well as for blood collection in cats undergoing cancer treatment. PMID- 11860244 TI - Secondary erythrocytosis associated with high plasma erythropoietin concentrations in a dog with cecal leiomyosarcoma. AB - A 14-year-old mixed-breed dog was examined because of severe absolute erythrocytosis (PCV, 70%). Plasma erythropoietin (EPO) concentration was consistently high, even though results of arterial blood gas analyses were normal. Radiography, ultrasonography, urinalysis, and serum biochemical analyses did not reveal any cardiac, pulmonary, or renal abnormalities that could cause the erythrocytosis, and erythrocytosis secondary to inappropriate EPO secretion was diagnosed. The PCV was maintained at approximately 60% by means of periodic phlebotomy, and the dog died of acute renal failure 2 years later. At necropsy, a cecal leiomyosarcoma was identified. Immunohistochemical staining of sections of the tumor revealed intracellular vacuoles containing EPO, and EPO mRNA was detected in the tumor by use of a reverse transcriptase-polymerase chain reaction assay These results suggested that ectopic production of EPO by a cecal leiomyosarcoma was the cause of erythrocytosis in this dog. PMID- 11860246 TI - Proceedings of the 12th International Symposium on Chiral Discrimination. France, 24-28 September 2000. PMID- 11860245 TI - Balloon valvuloplasty for palliative treatment of tricuspid stenosis with right to-left atrial-level shunting in a dog. AB - A 3.75-year-old castrated male Chesapeake Bay Retriever was referred for evaluation of tachypnea, exercise intolerance, and cyanosis. Echocardiographically, there was severe tricuspid stenosis and right-to-left atrial-level shunting of blood. Marked compensatory polycythemia had developed; the PCV was 75%. Balloon dilation of the tricuspid stenosis was performed. Subsequent echocardiographic examinations demonstrated a reduction in the pressure gradient across the tricuspid valve. The PCV returned to the reference range, and the dog's clinical status improved during the 12 months after the procedure. Tricuspid stenosis is an uncommon lesion in dogs and, in the dog of this report, was assumed to have resulted from tricuspid dysplasia. Cyanosis was a result of right-to-left shunting of blood. Limited treatment is available for dogs with cyanotic heart disease. In this dog, balloon dilation of the stenotic tricuspid valve was palliative. PMID- 11860247 TI - Efforts keep "pipeline" from drying up. PMID- 11860248 TI - "Pipeline of veterinarians drying up": ways to prime the pump. PMID- 11860249 TI - Correction--not opposed to USDA inspections. PMID- 11860251 TI - The rats, mice and birds issue has science advocacy "won" or "lost"? PMID- 11860252 TI - Alcohol as a disinfectant for aseptic surgery of rodents: crossing the thin blue line? AB - Recently, the use of alcohol (i.e., ethanol or isopropanol) has been discouraged as disinfectants for survival surgical procedures for rats and mice because of perceptions of inadequacy of killing of bacterial spores and lack of efficacy in the presence of organic debris. However, spore-forming bacteria are a minor (essentially nonexistent) threat for causing postoperative infections in rats and mice (but not necessarily hamsters, guinea pigs, or other rodents), and organic debris, with appropriate effort, can be removed from soiled instruments by using physical scrubbing. Although the metal-corrosive properties of alcohol and the lengthy times needed for adequate disinfection are drawbacks, there are essentially no microbiologically relevant reasons to discourage disinfection by ethanol or isopropanol in rat and mouse surgery. PMID- 11860253 TI - Individually ventilated cages: beneficial for mice and men? AB - Housing systems are an important element in the well-being of laboratory animals and, consequently, influence the outcomes of animal experiments. Individually ventilated cage (IVC) systems were developed to maintain low ammonia and CO(2) concentrations, to support a low relative humidity, and to reduce spread of infective agents and allergenic contaminants. However, the increased intracage ventilation rates (25 to 100 air changes/h) in these systems have animal welfare implications. In four preference studies involving a total of 36 pairs of female BALB/c mice and three different types of IVC racks, we tested the preference/avoidance of mice for the intracage ventilation rate, cage size, location of air supply, and presence of nesting material in a two-cage system. In this system, the two cages were connected by a passage with a swing-door to allow mice to move freely between the cages. We found that the mice avoided high intracage ventilation rates but that providing nesting material could counteract this avoidance. In addition, the mice preferred larger cages and an air supply in the cover. We conclude that the location of the air supply in the cage, cage size, ventilation rate and the presence of nesting material in IVC systems influence the well-being of the animals. PMID- 11860254 TI - Use of microisolator caging in a risk-based mouse import and quarantine program: a retrospective study. AB - The expanding use of genetically engineered mice has placed increasing demands upon research facilities to import mutant strains from noncommercial sources. Conventional quarantine strategies use relatively large amounts of space per animal and/or require specialized equipment (e.g., cubicles, isolators, and ventilated racks). We have retrospectively assessed a quarantine program instituted 4 years ago that used a small amount of space and minimized the need for special equipment. Shipments presumed, in light of health reports, to be free of agents excluded from our colonies were housed in static microisolator caging in a shared quarantine room. Rather than functioning as an "all-in-all-out" area, the room continually received new shipments, which were released intermittently as multi-shipment groups after testing was performed. Noninvasive testing of the imported mice was combined with nonsurvival sampling of sentinels that had been exposed to shipped animals via direct contact and/or exposure to soiled bedding. During the 4-year period examined, the vast majority of shipments presumed to be free of excluded agents showed no evidence of contamination when screened. When active infection was detected in the shared room, the procedures in place proved sufficient to prevent cross-contamination of other shipments. The use of sentinel animals to detect the shedding of infectious agents during quarantine was found to be an effective strategy that minimized the potential impact of invasive testing on quarantined animals. Although the program we describe is not appropriate for all situations, this type of approach may be considered by institutions wishing to explore alternatives to conventional quarantine strategies during periods of restricted space availability. PMID- 11860255 TI - Plasma homocysteine concentrations in a population of rhesus monkeys (Macaca mulatta): reference ranges and accompanying plasma concentrations of folate and vitamin B12. AB - The circulating level of homocysteine (Hcy) is recognized as a major independent risk factor for cardiovascular disease in humans. Nonhuman primates are being investigated to see whether they will be accurate models for Hcy indicators of cardiovascular dysfunction. Normal reference values are available in humans for Hcy as classified by age, gender, ethnic origin, and biological factors, however similar information in nonhuman primates had not been published previously. The purpose of this report is to provide normal Hcy values in a large group of nonhuman primates in light of age, gender, and physiologic state (pregnancy and lactation) and to compare these values to the same parameters in humans to highlight similar and dissimilar trends. In addition, plasma levels of folic acid and vitamin B(12), which are determinants of Hcy status in humans, are presented. Samples obtained from a troop of 149 rhesus monkeys (Macaca mulatta) fed a high protein commercial diet were analyzed for Hcy by using high-performance liquid chromatography. Folate and vitamin B(12) levels were determined by using an autoanalyzer. Results (mean [95% confidence interval]) for the entire troop were: Hcy, 4.5 (4.2-4.9) micromol/L; folic acid, 8.6 (8.0-9.1) micromol/L; and vitamin B(12), 673 (611-741) pmol/L. Quantitative values are similar to published values for another species of wild-caught macaques. Similar to trends noted for humans, male monkeys had higher Hcy values than did female animals, pregnant animals had lower values than did nonpregnant ones, and Hcy levels were inversely proportional to plasma folate and vitamin B(12) concentrations. However, homocysteine levels in rhesus monkeys did not vary consistently with age, whereas they increase with age in humans. PMID- 11860256 TI - A non-terminal surgical procedure for chronic collection of exocrine pancreatic secretions from unrestrained dogs (Canis familiaris). AB - The ability of dogs to adaptively modulate secretion by the exocrine pancreas to match changes in the amounts and sources of macronutrients is poorly understood. We evaluated the use of re-entrant pancreatic catheters as a non-terminal, temporary approach for the chronic collection of exocrine pancreatic secretion using unrestrained dogs fed diets differing in composition. Re-entrant catheters were surgically placed in the accessory pancreatic duct of two adult mongrel dogs. Secretions were collected for 40 days, during which the dogs were fed three diets with different amounts and sources of macronutrients. The volume of secretion was recorded, protein content was measured, and the activities of trypsin, chymotrypsin, amylase, and lipase were assayed. Inter-dog variation was detected for the volume of secretion (ml/h) but not for protein content (mg/ml) or activities (U/ml) of the enzymes. The volume and composition of the secretion differed among diets. The responses were delayed about 4 days, were transient, and did not coincide with the changes in diet composition. We found that the re entrant catheters were suitable for studying the exocrine pancreatic secretion of dogs. Our findings were inconclusive about the influence of diet but suggested that adult dogs have a limited and nonspecific response of pancreatic secretion. PMID- 11860257 TI - A minimally invasive percutaneous technique for jugular vein catheterization in pigs. AB - The objective of this study was to develop a simple, safe, reproducible, and efficient technique that used readily available commercial materials for percutaneous catheterization of an external jugular vein in the pig. Guidewire assisted vascular cannulation was introduced in the early 1950s and is often called the Seldinger technique, after its inventor. With few modifications, this technique has become widely used for all types of vascular cannulation in human and veterinary patients. The technique has the advantage of minimizing damage to soft tissues and blood vessels. We adapted this procedure by using five anatomical landmarks to target and catheterize the external jugular vein in pigs. Percutaneous catheterization of the external jugular vein can be accomplished easily in most pigs by using this technique. Novice technicians were able to quickly and easily learn the procedure. PMID- 11860258 TI - Otitis interna as a result of Pasteurella multocida infection in a laboratory woodchuck. AB - A laboratory woodchuck presented clinically with left-sided torticollis and a purulent exudate within the external auditory meatus of the left ear. Bacterial culture of the exudate resulted in a heavy growth of Pasteurella multocida. Treatment was initiated with topical and systemic antimicrobial compounds. There was no clinical improvement after 72 h of treatment, and euthanasia was elected. Radiographs correlated well with necropsy findings, confirming a diagnosis of otitis media; otitis interna was not confirmed but was suggested by the clinical presentation. To the authors knowledge, this is the first description of otitis media/interna as a result of P. multocida infection in a laboratory woodchuck. PMID- 11860259 TI - Chronic restraint via tail immobilization of mice: effects on corticosterone levels and other physiologic indices of stress. AB - Chronic venous cannulation in mice is an acceptable and useful technique for repeated blood sampling or continuous intravenous administration of substances, for which mild restraint of the animal may be necessary. Because chronic restraint has drawn considerable attention in the animal welfare community, the purpose of this study was to evaluate physiologic indices of stress in mice restrained by using an established tail restraint method. Serum corticosterone levels and body, thymus, adrenal, and spleen weights on days 2, 5, 8, and 12 were compared between tail-restrained and unrestrained mice. There were no significant differences between the two groups at the time points evaluated. Corticosterone levels were highest on day 8 for both groups and were significantly different from those on days 2 (P<0.009) and 5 (P<0.04) for restrained mice and on day 2 (P<0.02) for unrestrained mice. Levels in both groups declined by day 12, suggesting habitation. Weight loss was observed in all mice whether restrained or unrestrained. Significant differences in body, thymus, adrenal, and spleen weights were not evident between restrained and unrestrained mice. This study provides important information for balancing issues of prolonged restraint, animal well-being, and research goals. PMID- 11860260 TI - Perineal swellings in two strains of mice. AB - Seven adult male mice of two different strains and from two different animal housing facilities were submitted for necropsy to evaluate unilateral or bilateral swellings in the ventral perineal area. On dissection, the swellings were oval to spherical cystic structures located near the base of the penis. Striated muscle and a delicate stroma encapsulated all cystic structures. Cysts were firm, tan, and filled with clear, yellow fluid, or were softer and filled with white, amorphous, marbled material and a smaller amount of clear, yellow fluid. Evaluation of the cysts led us to a diagnosis of cystic bulbourethral (or Cowper s) glands. Possible causes and the incidence of this condition in other species are discussed. PMID- 11860261 TI - The Ustilaginales as plant pests and model systems. AB - The Ustilaginales are a vast and diverse group of fungi, which includes the plant pathogenic smuts that cause significant losses to crops worldwide. Members of the Ustilaginales are also valuable models for the unraveling of fundamental mechanisms controlling important biological processes. Ustilago maydis is an important fungal model system and has been well studied with regard to mating, morphogenesis, pathogenicity, signal transduction, mycoviruses, DNA recombination, and, recently, genomics. In this review we discuss the life cycles of members of the Ustilaginales and provide background on their economic impact as agricultural pests. We then focus on providing a summary of the literature with special attention to topics not well covered in recent reviews such as the use of U. maydis in mycovirus research and as a model for understanding the molecular mechanisms of fungicide resistance and DNA recombination and repair. PMID- 11860262 TI - Cloning of glyceraldehyde-3-phosphate dehydrogenase-encoding genes in Mucor circinelloides (Syn. racemosus) and use of the gpd1 promoter for recombinant protein production. AB - Three genes (gpd1, gpd2, and gpd3) encoding glyceraldehyde-3-phosphate dehydrogenase were isolated from the dimorphic zygomycete Mucor circinelloides by PCR using degenerated primers. Transcription of gpd1 could be detected during vegetative growth under both aerobic and anaerobic conditions, whereas neither gpd2 nor gpd3 transcription was detected, indicating that gpd1 is the major transcribed gpd gene. The transcription of gpd1 was regulated by carbon source. The gpd1 promoter was successfully used for recombinant expression of genes of both homologous (crgA encoding a regulator of carotene biosynthesis) and heterologous (gox1 from Aspergillus niger encoding glucose oxidase; GOX) nature. Growth of a gox1 transformant strain resulted in the secretion of enzymatically active GOX. The potential advantages of using a dimorphic fungus for heterologous protein production are discussed. PMID- 11860263 TI - An extended physical map of the TOX2 locus of Cochliobolus carbonum required for biosynthesis of HC-toxin. AB - In genetic crosses, HC-toxin production in the filamentous fungus Cochliobolus carbonum appears to be controlled by a single locus, TOX2. At the molecular level, TOX2 is composed of at least seven duplicated and coregulated genes involved in HC-toxin biosynthesis, export, and regulation. All copies of four of the TOX2 genes were previously mapped within a 540-kb stretch of DNA in strain SB111. Subsequently, an additional three TOX2 genes, TOXE, TOXF, and TOXG, have been discovered. In this paper we have mapped all copies of the new genes, a total of seven, and show that except for one of the two copies of TOXE, which was previously shown to be on a chromosome of 0.7 Mb in strain SB111, they are all linked to the previously known TOX2 genes within approximately 600 kb of each other on a chromosome of 3.5 Mb. We show here that this chromosome also contains at least one non-TOX2 gene, EXG2, which encodes an exo-beta1,3-glucanase. EXG2 is still present in strains that have undergone spontaneous deletion of up to approximately 1.4 Mb of the 3.5-Mb chromosome. The results contribute to our understanding of the complex organization of the genes involved in HC-toxin biosynthesis and are consistent with the hypothesis that a reciprocal chromosomal translocation accounts for the pattern of distribution of the TOX2 genes in different C. carbonum isolates. PMID- 11860264 TI - Concerted evolution in the ribosomal RNA genes of an Epichloe endophyte hybrid: comparison between tandemly arranged rDNA and dispersed 5S rrn genes. AB - We examined ribosomal RNA concerted evolution in an Epichloe endophyte interspecific hybrid (Lp1) and its progenitors (Lp5 and E8). We show that the 5S rrn genes are organized as dispersed copies. Cloned 5S gene sequences revealed two subfamilies exhibiting 12% sequence divergence, with substitutions forming coevolving pairs that maintain secondary structure and presumably function. Observed sequence patterns are not fully consistent with either concerted or classical evolution. The 5S rrn genes are syntenic with the tandemly arranged rDNA genes, despite residing outside the rDNA arrays. We also examined rDNA concerted evolution. Lp1 has rDNA sequence from only one progenitor and contains multiple rDNA arrays. Using 5S rrn genes as chromosomal markers, we propose that interlocus homogenization has replaced all Lp5 rDNA sequence with E8 sequence in the hybrid. This interlocus homogenization appears to have been rapid and efficient and is the first demonstration of hybrid interlocus homogenization in the Fungi. PMID- 11860265 TI - Cloning and characterization of a cDNA of cro rI from the white pine blister rust fungus Cronartium ribicola. AB - White pine blister rust (WPBR) is caused by the fungus Cronartium ribicola which has five spore stages on two unrelated hosts, the five-needle pines and Ribes spp. Recently, during the molecular analysis of the proteins and genes involved in host-pathogen interaction, the WPBR fungal protein Cro rI was identified in infected white pine tissues. To further characterize Cro rI, an expression cDNA library from poly(A)(+) mRNA of C. ribicola axenic mycelial culture was constructed and immunoscreened and the cDNA was cloned. Sequence analysis indicated an open reading frame of 462 bases, which encodes a protein of 153 amino acid residues with a molecular mass of 16.7 kDa and a predicted isoelectric point (pI) of 8.93. Based on the N-terminal amino acid sequences of Cro rI, the secreted portion of Cro rI protein should be 136 amino acids long with several putative posttranslational modification sites and a molecular mass of 14.8 kDa. The predicted pI for the secreted portion was 9.34. The predicted N-terminal signal peptide was 17 amino acids long. The N-terminal 42-amino acid sequence of the predicted mature protein (secreted portion) was identical to the amino terminal sequence of Cro rI that was previously determined. Southern blot hybridizations indicated that the C. ribicola genome contained at least two copies of the cro rI gene. Isolation of the genomic PCR fragment, which was approximately 400 bp longer than the cDNA, and subsequent cloning and sequencing analyses confirmed that there were three introns within the coding regions. Western immunoblot analyses revealed that Cro rI protein accumulated in large amounts only in the infected white pine tissues while no trace was detectable in the alternate Ribes stage or the five different spores, suggesting a critical role of Cro rI in the haploid stage of the fungus (in pine). The translocation of Cro rI was only found to occur in cankered trees, and not in the young infected seedlings. The implications of Cro rI in pathogenesis are discussed. PMID- 11860266 TI - Purification and characterization of chitinases from the nematophagous fungi Verticillium chlamydosporium and V. suchlasporium. AB - Culture filtrates of the nematophagous fungi Verticillium chlamydosporium and V. suchlasporium growing on colloidal chitin showed increasing chitinolytic activity and production of two (32- and 43-kDa) main proteins. Maximum activity was found 18-20 days after inoculation, but V. suchlasporium always displayed higher activity. Zymography of such filtrates on carboxymethyl-chitin-Remazol brilliant violet 5R/acrylamide gels showed five bands of substrate degradation for V. suchlasporium and three for V. chlamydosporium. Filtrates with maximum activity were chromatographed on macroporous cross-linked chitin affinity matrix, showing a peak of main (50-60%) activity, which only contained a 43-kDa protein for both fungi. Zymography and colloidal chitin degradation showed that it was a single endochitinase (CHI43) with optimum pH range of 5.2-5.7. The main isoforms had pIs of 7.6 for V. suchlasporium and 7.9 for V. chlamydosporium. Eggs of the nematode Globodera pallida treated with CHI43 and the serine protease P32 from V. suchlasporium alone or in combination showed surface damage in comparison with controls when examined by scanning electron microscopy. PMID- 11860267 TI - Electrical muscle activity pattern and transcriptional and posttranscriptional mechanisms regulate PKA subunit expression in rat skeletal muscle. AB - We have examined protein kinase A (PKA) subunit expression in adult rat skeletal muscles. Northern blots identified PKA catalytic alpha and regulatory (R) I alpha and RII alpha subunits as the major subunits expressed in slowly contracting soleus (SOL) and rapidly contracting extensor digitorum longus (EDL) muscles. In addition, the steady-state RNA levels of PKA subunit mRNAs and activities of RI alpha and RII alpha promoters are similar in SOL and EDL. These data indicate that posttranscriptional mechanisms account for the twofold differences in PKA subunit protein levels reported earlier. Electrical stimulation of denervated SOL with an EDL-like activity pattern (fast pattern) transformed SOL into an EDL-like muscle with regard to PKA protein levels. These experiments suggest that the posttranscriptional regulation is activity pattern-dependent. Denervation specifically increased RI alpha promoter activity and RI alpha mRNA levels in SOL and EDL. Further experiments indicated that the RI alpha 1a upstream sequences were activated following denervation. Direct electrical stimulation prevented the rise in RI alpha mRNA levels following denervation, demonstrating that electrical muscle activity regulates transcription. PMID- 11860268 TI - Reduced cortical synaptic plasticity and GluR1 expression associated with fragile X mental retardation protein deficiency. AB - Lack of expression of the fragile X mental retardation protein (FMRP), due to silencing of the FMR1 gene, causes the Fragile X syndrome. Although FMRP was characterized previously to be an RNA binding protein, little is known about its function or the mechanisms underlying the Fragile X syndrome. Here we report that the alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate receptor subunit, GluR1, was decreased in the cortical synapses, but not in the hippocampus or cerebellum, of FMR1 gene knockout mice. Reduced long-term potentiation (LTP) was also found in the cortex but not in the hippocampus. Another RNA binding protein, FXR; the N-methyl-D-aspartate receptor subunit, NR2; and other learning-related proteins including c-fos, synapsin, myelin proteolipid protein, and cAMP response element binding protein were not different between FMR1 gene knockout and wild type mice. These findings suggest that the depressed cortical GluR1 expression and LTP associated with FMRP deficiency could contribute to the Fragile X phenotype. PMID- 11860269 TI - The amino terminus of the glial glutamate transporter GLT-1 interacts with the LIM protein Ajuba. AB - We have identified a cytoplasmic LIM protein, Ajuba, which interacts with the amino terminus of GLT-1, the most abundant plasma membrane glutamate transporter in the brain. Ajuba has a cytoplasmic location when expressed alone in COS cells, but translocates to colocalize with GLT-1 at the plasma membrane when GLT-1 is coexpressed. Ajuba is expressed in cerebellum, cortex, hippocampus, and retina and also in organs outside the CNS. Ajuba is found with GLT-1 in astrocytes, cerebellar Bergmann glia and retinal neurons, and antibodies to Ajuba coimmunoprecipitate GLT-1 from brain. For GLT-1 expressed in COS cells, coexpression of Ajuba did not affect the transporter's K(m) or V(max) for glutamate. Since Ajuba is known to activate MAP kinase enzymes, and its homologue Zyxin binds to cytoskeletal proteins, we propose that Ajuba is a scaffolding protein allowing GLT-1 to regulate intracellular signaling or interact with the cytoskeleton. PMID- 11860270 TI - Preferential transfection of adult mouse neural stem cells and their immediate progeny in vivo with polyethylenimine. AB - The subventricular zone of the adult mammalian brain harbors the neural stem cell population with potential neural regeneration and repair capacity. We describe a nonviral technique to preferentially transfect in vivo the adult neural stem cell population and its immediate progeny based on intraventricular injection of PEI/DNA complexes. The transfected population was identified by cellular and ultra-structural evidence showing their proliferating status and expression of the specific markers GFAP and nestin. Stable activation of the lacZ reporter by cre-recombinase transfection in R26R mice demonstrated survival and migration of stem cell derivatives three months after injection. Apoptosis is thought to be the most common fate of the stem cell progeny. Overexpression of Bcl-X(L) increased number and survival time of transduced progenitors and decreased the frequency of cells immunopositive for activated Caspase-3. This method thus provides selective targeting of the stem cell population and should allow an in depth understanding of their biology. PMID- 11860271 TI - The carboxyl-terminus of BACE contains a sorting signal that regulates BACE trafficking but not the formation of total A(beta). AB - BACE (beta-site APP cleaving enzyme) has been recently proposed as the major aspartyl protease displaying beta secretase activity in neurons. The C-terminal domain of BACE contains a dileucine motif (LL499/500) that can potentially regulate its trafficking and endocytosis, and an adjacent serine, which is a potential phosphorylation site (S498) that could modulate the activity of the LL motif. In this paper we show that S498 is phosphorylated by casein kinase 1 (CKI). Mutating the LL to dialanine (AA) caused an increase in the levels of mature BACE. The LL to AA mutation increased levels of BACE on the cell surface and decreased the internalization of BACE. Mutating the S498 to alanine did not alter levels of cell surface BACE. Mutating either the leucines or the serine did not alter the secretion of A(beta). Our data are consistent with a role for the cytoplasmic domain in regulating BACE trafficking and localization. PMID- 11860272 TI - Upregulation of bradykinin B2 receptor expression by neurotrophic factors and nerve injury in mouse sensory neurons. AB - Bradykinin B2 receptor mRNA was detected at low levels, both by RT-PCR and by in situ hybridization, in freshly isolated dorsal root ganglia (DRG) and in ganglia cultured in the absence of neurotrophic factors, but was strongly upregulated by culture in the presence of nerve growth factor (NGF). The effect of NGF is mediated via TrkA receptors. The related neurotrophins, brain-derived neurotrophic factor, neurotrophin-3, and neurotrophin-4, were ineffective in upregulating B2 mRNA, but a small upregulation was seen with the unrelated neurotrophin glial cell line-derived neurotrophic factor (GDNF). Surface membrane B2 receptor expression, detected by immunofluorescence using a B2-specific antibody, was low in outgrowing axons cultured in the absence of neurotrophic factors, but was elevated by addition of NGF or GDNF. Conditioned media prepared by incubating injured nerve, skin, or muscle had a similar effect to NGF in upregulating B2 mRNA and protein expression, and the activity was largely removed by neutralization of NGF in the conditioned medium with an anti-NGF antibody. After nerve crush injury in vivo an enhancement in B2 mRNA expression was seen, peaking after 7 days and returning to precrush levels after 14 days. In all conditions tested, the proportion of neurons expressing B2 mRNA remained the same at around 23% of small neurons, suggesting that upregulation only occurs in the B2-positive neurons. These experiments show that NGF, and to a lesser extent GDNF, upregulates the expression of bradykinin B2 mRNA and B2 receptor protein in the surface membrane of DRG neurons and that NGF is an important factor responsible for upregulating bradykinin B2 receptor expression after nerve crush injury in vivo. PMID- 11860273 TI - Strychnine-blocked glycine receptor is removed from synapses by a shift in insertion/degradation equilibrium. AB - The long-term inhibition by strychnine of glycine receptor activity in neurons provokes the receptor's selective intracellular accumulation and disappearance from synapses. This could result either from a disruption of the postsynaptic anchoring of the receptor or from an arrest of its exocytic transport. In this study we combined biochemical and fluorescence microscopy analyses to determine on a short time scale the fate of the strychnine-inactivated glycine receptor. Quantification of the cellular content of receptor showed that the rapid accumulation depends on protein synthesis. Cell surface biotinylation of neurons demonstrated that strychnine did not accelerate the turnover rate of the receptor. Labeling of endosomes indicated that, in strychnine-treated cells, the accumulated receptor is not blocked in the endosomal transport pathway. Taken together, these results indicate that strychnine does not destabilize the postsynaptic receptor but triggers its disappearance from synapses by a nondegradative sequestration of newly synthesized molecules in a nonendocytic compartment. PMID- 11860274 TI - Loss of prion protein in a transgenic model of amyotrophic lateral sclerosis. AB - Amyotrophic lateral sclerosis (ALS) is a motor neuron degenerative disorder caused in a proportion of cases by missense mutations in the gene encoding Cu/Zn superoxide dismutase (Cu/Zn-SOD) which result in unknown, lethal enzymatic activity. Based on a differential screening approach, we show here that the gene encoding the cellular prion protein (PrP(C)) was specifically repressed in a transgenic model of ALS overexpressing the mutant G86R Cu/Zn-SOD. Analysis by Northern blot, semiquantitative RT-PCR, and Western blot revealed that PrP(C) down-regulation, which appeared early in the asymptomatic phase of the pathology, occurred preferentially in those tissues primarily affected by the disease (spinal cord, sciatic nerve, and gastrocnemius muscle). This down-regulation was not accompanied by refolding of the aberrant PrP(Sc) isoform, the agent which causes transmissible spongiform encephalopathies. Furthermore, modification of PrP(C) expression was specifically linked to the presence of the G86R mutant since no changes were observed in transgenic mice overexpressing wild-type Cu/Zn SOD. PrP(C) has been shown to play a role in the protection against oxidative stress, and we therefore propose that its down-regulation may contribute at least in part to ALS pathogenesis. PMID- 11860275 TI - Loss of polysialic residues accelerates CNS neural precursor differentiation in pathological conditions. AB - Using the model of lysolecithin-induced demyelination of the corpus callosum in wild-type, NCAM-deficient, and endoneuraminidase-injected mice, we have analyzed the consequences of the loss of expression of NCAM or PSA residues on the migration and proliferation capacities of neural precursors of the subventricular zone (SVZ). We showed that the absence of PSA or NCAM delayed migration of neural precursors to the olfactory bulb and consequently enhanced their recruitment at the lesion site. Moreover, after demyelination, the lack of NCAM but not PSA promoted proliferation in the SVZ and the lesion while the lack of PSA favored the differentiation of the traced cells into the oligodendroglial fate both in the SVZ and in the lesion. As previously demonstrated in vitro (L. Decker et al., 2000, Mol. Cell. Neurosci. 16, 422-439), these data illustrate the involvement of PSA and NCAM in neural precursor motility and differentiation in the normal and injured central nervous system, suggesting distinct roles for these two molecules under pathophysiological conditions. PMID- 11860276 TI - Neurotrophin-3-mediated regeneration and recovery of proprioception following dorsal rhizotomy. AB - Injured dorsal root axons fail to regenerate into the adult spinal cord, leading to permanent sensory loss. We investigated the ability of intrathecal neurotrophin-3 (NT3) to promote axonal regeneration across the dorsal root entry zone (DREZ) and functional recovery in adult rats. Quantitative electron microscopy showed robust penetration of CNS tissue by regenerating sensory axons treated with NT3 at 1 and 2 weeks postrhizotomy. Light and electron microscopical anterograde tracing experiments showed that these axons reentered appropriate and ectopic laminae of the dorsal horn, where they formed vesicle-filled synaptic buttons. Cord dorsum potential recordings confirmed that these were functional. In behavioral studies, NT3-treated (but not untreated or vehicle-treated) rats regained proprioception. Recovery depended on NT3-mediated sensory regeneration: preventing regeneration by root excision prevented recovery. NT3 treatment allows sensory axons to overcome inhibition present at the DREZ and may thus serve to promote functional recovery following dorsal root avulsions in humans. PMID- 11860277 TI - Down-regulation of the retinoblastoma protein (rb) is associated with rat oligodendrocyte differentiation. AB - Terminal differentiation of oligodendrocytes is associated with permanent withdrawal from the cell cycle. We studied the expression of the retinoblastoma protein, expression and activity of G1 cyclins and kinases in oligodendrocyte progenitor cells cultured in vitro. We found that Rb stopped to be expressed concomitantly with the activation of CNPase in oligodendrocytes differentiated with thyroid hormone. In contrast, Rb continued to be expressed at reduced levels in oligodendrocytes that were arrested in G1 by removal of mitogens. Cyclin D1, cdk2, and cdk4 kinase activities were decreased in G1-arrested and differentiated oligodendrocytes. Cyclin E, however, continued to be expressed in G1-arrested oligodendrocytes. Inhibition of differentiation induced by mitogens in oligodendrocytes arrested in G1 by Ad-p27 was accompanied by continued expression of Rb, D1, and E cyclins. After removal of mitogens and addition of thyroid hormone, Rb stopped being expressed and CNPase expression was activated with a temporal course similar to that of oligodendrocytes infected with a control adenovirus. Our results indicate that Rb may play an important function in differentiation of oligodendrocytes in response to external mitogens and differentiation factors. PMID- 11860278 TI - Dopamine D2 receptor signaling controls neuronal cell death induced by muscarinic and glutamatergic drugs. AB - Dopamine (DA), through D1/D2 receptor-mediated signaling, plays a major role in the control of epileptic seizures arising in the limbic system. Excitotoxicity leading to neuronal cell death in the affected areas is a major consequence of seizures at the cellular level. In this respect, little is known about the role of DA receptors in the occurrence of epilepsy-induced neuronal cell death. Here we analyze the occurrence of seizures and neurotoxicity in D2R -/- mice treated with the cholinergic agonist pilocarpine. We compared these results with those previously obtained with kainic acid (KA), a potent glutamate agonist. Importantly, D2R -/- mice develop seizures at doses of both drugs that are not epileptogenic for WT littermates and show greater neurotoxicity. However, pilocarpine-induced seizures result in a more widespread neuronal death in both WT and D2R -/- brains in comparison to KA. Thus, the absence of D2R lowers the threshold for seizures induced by both glutamate and acetylcholine. Moreover, the dopaminergic control of epilepsy-induced neurodegeneration seems to be mediated by distinct interactions of D2R signaling with these two neurotransmitters. PMID- 11860279 TI - Purinergic (P2X7) receptor activation of microglia induces cell death via an interleukin-1-independent mechanism. AB - Activation of purinergic P2X7 receptors, principally by extracellular ATP, promotes the processing and release of the cytokine interleukin-1beta (IL-1beta) and induces cell death in activated microglia and macrophages. The objective of this study was to determine if IL-1beta release contributes directly to this cell death in microglia. Exposure of microglia to bacterial lipopolysaccharide (LPS) and ATP induced release of IL-1beta and IL-1alpha, as well as cell death. Neither cell death nor IL-1 release was observed in microglia lacking the P2X7 receptor. Microglia from mice lacking the IL-1beta gene demonstrated a profile of death identical to that of wild-type microglia in response to LPS and ATP. Thus, IL 1beta is not required for P2X7 receptor-stimulated microglial death. PMID- 11860280 TI - High extracellular potassium protects against the toxicity of cytosine arabinoside but is not required for the survival of cerebellar granule cells in vitro. AB - Depolarization of cerebellar granule cells with elevated potassium has been described as essential to maintain their survival in culture. There are several reports that this is only specific for rat cerebellar granule cells and not those of mouse. We reinvestigated this issue and found that although high potassium enhanced the survival of cerebellar granule cells from both rat and mouse it was not essential for the survival of those cultures. Further analysis of the culture system indicated that high potassium offered protection against the toxicity of glutamate and cytosine arabinose (Ara C), a standard antimitotic additive to cultures of granule cells. Ara C was found to be toxic to cerebellar cells after potassium withdrawal at concentrations standardly used in culturing these cells (10 microM). High potassium was found to diminish the expression of p53. Ara C toxicity is known to utilize the p53-dependent signaling pathway to initiate apoptosis. Another depolarizing agent, veratridine, offers no protection against Ara C but we provide evidence that the protective effect of high potassium against Ara C is mediated through calcium balance within the cells. We suggest that there is no requirement for high potassium in terms of cerebellar granule cell survival. The previously proposed role for high potassium in the survival cerebellar granule cells is rather a protective effect against toxic substances in serum such as glutamate or against agents such as Ara C. PMID- 11860281 TI - Protein kinase C delta (PKCdelta) is required for protein tyrosine phosphatase mu (PTPmu)-dependent neurite outgrowth. AB - Protein tyrosine phosphatase mu (PTPmu) is an adhesion molecule in the immunoglobulin superfamily and is expressed in the developing nervous system. We have shown that PTPmu can promote neurite outgrowth of retinal ganglion cells and it regulates neurite outgrowth mediated by N-cadherin (S. M. Burden-Gulley and S. M. Brady-Kalnay, 1999, J. Cell Biol. 144, 1323-1336). We previously demonstrated that PTPmu binds to the scaffolding protein RACK1 in yeast and mammalian cells (T. Mourton et al., 2001, J. Biol. Chem. 276, 14896-14901). RACK1 is a receptor for activated protein kinase C (PKC). In this article, we demonstrate that PKC is involved in PTPmu-dependent signaling. PTPmu, RACK1, and PKCdelta exist in a complex in cultured retinal cells and retinal tissue. Using pharmacologic inhibition of PKC, we demonstrate that PKCdelta is required for neurite outgrowth of retinal ganglion cells on a PTPmu substrate. These results suggest that PTPmu signaling via RACK1 requires PKCdelta activity to promote neurite outgrowth. PMID- 11860283 TI - Ribosomal crystallography: from poorly diffracting microcrystals to high resolution structures. AB - The cellular organelles translating the genetic code into proteins, the ribosomes, are large, asymmetric, flexible, and unstable ribonucleoprotein assemblies, hence they are difficult to crystallize. Despite two decades of intensive effort and thorough searches for suitable sources, so far only three crystal types have yielded high-resolution structures: two large subunits (from an archaean and from a mesophilic eubacterium) and one thermophilic small subunit. These structures have added to our understanding of decoding, have revealed dynamic aspects of the biosynthetic process, and have indicated the strategies adopted by ribosomes for interacting between themselves as well as with inhibitors, factors and substrates. PMID- 11860284 TI - Construction of low-resolution x-ray crystallographic electron density maps of the ribosome. AB - Advances in X-ray crystallography now allow biological macromolecules of almost any size to be imaged at atomic resolution. Here, I outline the strategy that allowed for the solution of the 70S ribosome structure to 7.8-A resolution. The most important factors involve the effective use of synchrotron radiation and the application of existing crystallographic software to very large structures. PMID- 11860285 TI - Ribosomal dynamics explored by cryo-electron microscopy. AB - Cryo-electron microscopy has furnished direct evidence for conformational changes of the ribosome as it proceeds, in a cyclic manner, through different functional states. Strategies to explore the ribosome dynamics include trapping of particular functionally meaningful states by chemical, genetic, or physical means. The new atomic information obtained by X-ray crystallography should make it possible to track conformational changes observed by cryo-electron microscopy to changes of the underlying structural framework of the ribosome. PMID- 11860286 TI - RNAPack: an integrated NMR approach to RNA structure determination. AB - Over the last decade, a vast number of useful nuclear magnetic resonance (NMR) experiments have been developed and successfully employed to determine the structure and dynamics of RNA oligonucleotides. Despite this progress, high resolution RNA structure determination by NMR spectroscopy still remains a lengthy process and requires programming and extensive calibrations to perform NMR experiments successfully. To accelerate RNA structure determination by NMR spectroscopy, we have designed and programmed a package of RNA NMR experiments, called RNAPack. The user-friendly package contains a set of semiautomated single, double, and triple resonance NMR experiments, which are fully optimized for high resolution RNA solution structure determination on Varian NMR spectrometers. RNAPack provides an autocalibration feature that allows rapid calibration of all NMR experiments in a single step and thereby speeds up the NMR data collection and eliminates user errors. In our laboratory, we have successfully employed this technology to solve RNA solution structures of domains of the internal ribosome entry site of the genomic hepatitis C viral RNA in less than 3 months. RNAPack therefore makes NMR spectroscopy an attractive and rapid structural tool and allows integration of atomic resolution structural information into biochemical studies of large RNA systems. PMID- 11860287 TI - Short-range RNA-RNA crosslinking methods to determine rRNA structure and interactions. AB - We describe details of procedures to analyze RNA-RNA crosslinks made by far-UV irradiation (< 300 nm) or made by irradiation with near-UV light (320-365 nm) on RNA containing photosensitive nucleotides, in the present case containing 4 thiouridine. Zero-length crosslinks of these types must occur because of the close proximity of the participants through either specific interactions or transient contacts in the folded RNA structure, so they are valuable monitors of the conformation of the RNA. Procedures to produce crosslinks in the 16S ribosomal RNA and between the 16S rRNA and mRNA or tRNA are described. Gel electrophoresis conditions are described that separate the products according to their structure to allow the determination of the number and frequency of the crosslinking products. Gel electrophoresis together with an ultracentrifugation procedure for the efficient recovery of RNA from the polyacrylamide gels allows the purification of molecules containing different crosslinks. These separation techniques allow the analysis of the sites of crosslinking by primer extension and RNA sequencing techniques. The procedures are applicable to other types of RNA molecules with some differences to control levels of crosslinking and separation conditions. PMID- 11860288 TI - Comparison of rRNA cleavage by complementary 1,10-phenanthroline-Cu(II)- and EDTA Fe(II)-derivatized oligonucleotides. AB - The chemical nucleases 1,10-phenanthroline-Cu(II) and EDTA-Fe(II), have proven to be valuable tools for structural analysis of nucleic acids. Both have found applications in footprinting and directed proximity studies of DNA and RNA. Derivatives of each that provide for tethering to nucleic acid or protein are commercially available, allowing their widespread use for structural analysis of macromolecules. Although their applications are somewhat overlapping, differences in their cleavage mechanisms and chemical properties allow them to provide distinct and complementary structural information. The purpose of this study is to compare directly the cleavage patterns of tethered 1,10-phenanthroline-Cu(II) and EDTA-Fe(II) complexes within a similar experimental system. Here, the region surrounding nucleotide 1400 of 16S rRNA from Escherichia coli serves as a substrate for chemical cleavage directed by a derivatized complementary oligonucleotide. This region of rRNA is known to be involved in the decoding of mRNA during translation. The results of this study provide evidence in support of the mechanistic differences previously established for EDTA-Fe(II) and 1,10 phenathroline-Cu(II). The delocalized cleavage envelope produced by EDTA-Fe(II) cleavage suggests the involvement of a diffusible reactive species. On the other hand, rRNA cleavage induced by the tethered 1,10-phenanthroline-Cu(II) complex appears localized to the proximity of the chemical nuclease under normal conditions, although the production of an unknown diffusible species appears to occur during long reaction times. PMID- 11860289 TI - SERF: in vitro election of random RNA fragments to identify protein binding sites within large RNAs. AB - In vitro selection experiments have various goals depending on the composition of the initial pool and the selection method applied. We developed an in vitro selection variant (SERF, selection of random RNA fragments) that is useful for the identification of short RNA fragments originating from large RNAs that bind specifically to a protein. A pool of randomly fragmented RNA is constructed from a large RNA, which is the natural binding partner for a protein. Such a pool contains all the potential binding sites and is therefore used as starting material for affinity selection with the purified protein to find its natural target. Here we provide a detailed experimental protocol of the method. SERF has been developed for ribosomal systems and is a general approach providing a basis for functional and structural characterization of RNA-protein interactions in large ribonucleoprotein particles. PMID- 11860290 TI - In vivo selection of functional variations in essential sites of ribosomal RNA. AB - The technique of "in vivo selection of functional ribosomes" is a genetic approach to dissecting the link between the structure and function of critical sites of rRNA. This method proceeds through selection of functional variants among cells that express ribosomes from a pool of rRNA-containing randomized sites. The selection of bacterial clones with functional ribosomes is based on the use of a plasmid carrying a rRNA operon in which a site of interest has been randomized and a point mutation conferring an antibiotic resistance has been introduced. Cells expressing functional ribosomes are then selected on medium containing the antibiotic. With this approach one can isolate at once all the possible variations at a given rRNA site that are able to sustain normal ribosome function. The identification of covariations in between several nucleotides that maintain wild-type ribosome activity can thus help demonstrate the function of specific interactions in rRNA. PMID- 11860291 TI - Mapping pseudouridines in RNA molecules. AB - Pseudouridine is present in ribosomal RNA, transfer RNA, tmRNA, and small nuclear and nucleolar RNAs. All are structured molecules. Pseudouridine is made by enzyme catalyzed isomerization of specifically selected U residues after the polynucleotide chain is made. No energy input is required. Pseudouridine formation creates a hydrogen bond donor at the equivalent of uridine C-5. Therefore, a major role of pseudouridine may be to strengthen particular RNA conformations and/or RNA-RNA interactions because of this extra H-bond capability. Understanding the role of pseudouridine critically depends on knowledge of their location and number in RNA. The mapping method described here has greatly simplified this task and made it possible to survey many organisms. Procedures are described for mapping pseudouridines in large RNAs like ribosomal RNA and in small RNAs like tRNA. The method involves carbodiimide adduct formation with U, G, and pseudouridine followed by mild alkali to remove the adduct from U and G but not from the N-3 of pseudouridine. This results in attenuation of primed reverse transcription resulting in a stop band one residue 3' to the pseudouridine on sequencing gels. Use of primers means that purified RNAs are not needed, only knowledge of its primary sequence, but limit the sequence scanned to 30-40 residues from the 3' end. A poly(A) tailing procedure is described that allows extension of the method to within a few nucleotides of the 3' terminus. PMID- 11860293 TI - Staining of sentinel node reticulum cells with cytokeratin antibody AE3. PMID- 11860292 TI - Mapping 2'-O-methyl groups in ribosomal RNA. AB - Ribosomal RNAs (rRNAs) from all sources contain modified nucleosides, whose numbers range from a few in mitochondrial rRNA to more than 200 in the complete rRNAs of some higher eukaryotes. In eukaryotic rRNA the great majority of modified nucleosides are 2'-O-methylated nucleosides or pseudouridines. The locations of most of the 2'-O-methylated nucleosides in rRNA from some representative eukaryotes are known from studies whose aim was full characterization of rRNA methylation. More recently, and particularly in connection with the discovery of methylation guide RNAs, it is often required to check for the presence or absence of 2'-O-methyl nucleosides at specified locations within rRNA. Three methods that can be applied for such "local" objectives are reviewed. Two of the methods are based on primer extension by reverse transcriptase. They exploit, respectively, a tendency of 2'-O-methyl groups to impede reverse transcriptase at low dNTP concentrations, or the resistance of phosphodiester bonds adjacent to 2'-O-methyl groups to alkaline hydrolysis. Examples of these methods are summarized. Although the two methods are relatively straightforward, they suffer from various experimental limitations, as discussed. The third method is technically more sophisticated but is capable of overcoming the limitations of the first two methods. It is based on the resistance of a target 2'-O-methylated site to cleavage by RNase H when the site is hybridized to an appropriate chimeric oligonucleotide. An overview of the approaches and methods now available for the complete mapping of 2'-O-methyl groups in rRNA is presented. PMID- 11860295 TI - Pierre Paul Broca. PMID- 11860296 TI - The postgenomic era: implications for the clinical laboratory. AB - OBJECTIVES: To review the advances in clinically useful molecular biological techniques and to identify their applications in clinical practice, as presented at the Tenth Annual William Beaumont Hospital DNA Symposium. DATA SOURCES: The 11 manuscripts submitted were reviewed and their major findings were compared with literature on the same topic. STUDY SELECTION: Manuscripts address creative thinking techniques applied to DNA discovery, extraction of DNA from clotted blood, the relationship of mitochondrial dysfunction in neurodegenerative disorders, and molecular methods to identify human lymphocyte antigen class I and class II loci. Two other manuscripts review current issues in molecular microbiology, including detection of hepatitis C virus and biological warfare. The last 5 manuscripts describe current issues in molecular cardiovascular disease, including assessing thrombotic risk, genomic analysis, gene therapy, and a device for aiding in cardiac angiogenesis. DATA SYNTHESIS: Novel problem solving techniques have been used in the past and will be required in the future in DNA discovery. The extraction of DNA from clotted blood demonstrates a potential cost-effective strategy. Cybrids created from mitochondrial DNA depleted cells and mitochondrial DNA from a platelet donor have been useful in defining the role mitochondria play in neurodegeneration. Mitochondrial depletion has been reported as a genetically inherited disorder or after human immunodeficiency virus therapy. Hepatitis C viral detection by qualitative, quantitative, or genotyping techniques is useful clinically. Preparedness for potential biological warfare is a responsibility of all clinical laboratorians. Thrombotic risk in cardiovascular disorders may be assessed by coagulation screening assays and further defined by mutation analysis for specific genes for prothrombin and factor V Leiden. Gene therapy for reducing arteriosclerotic risk has been hindered primarily by complications introduced by the vectors used to introduce the therapeutic genes. Neovascularization in cardiac muscle with occluded vessels represents a promising method for recovery of viable tissue following ischemia. CONCLUSIONS: The sequence of the human genome was reported by 2 groups in February 2001. The postgenomic era will emphasize the use of microarrays and database software for genomic and proteomic screening in the search for useful clinical assays. The number of molecular pathologic techniques and assays will expand as additional disease-associated mutations are defined. Gene therapy and tissue engineering will represent successful therapeutic adjuncts. PMID- 11860297 TI - Mental tools for thinking about DNA technologies in new ways. AB - OBJECTIVE: To investigate the nature of creative thinking in biomedical science with specific applications to molecular pathologies and DNA technologies. DATA SOURCES: Accounts of breakthroughs and inventions contained in autobiographies, biographies, interviews, and archival sources. STUDY SELECTION: Discoveries that have altered, or may yet alter, basic textbook accounts of biomedical sciences for which appropriate data sources exist. DATA EXTRACTION: Approximately 1000 data sources were analyzed, both within appropriate sciences and in other creative fields, such as the arts. DATA SYNTHESIS: The current analysis is based on a framework described in our previous book, Sparks of Genius, which outlines a general approach to understanding creative thinking. CONCLUSIONS: Creative thinking in all disciplines depends on a common mental "toolkit" that consists of 13 fundamental tools: observing, imaging, abstracting, pattern recognition, pattern forming, analogizing, body thinking, empathizing, dimensional thinking, modeling, playing, transforming, and synthesizing. Scientists recognize and solve problems by observing data that break the patterns established by theories; exploring a system by creating an abstract model with which they can play; and transforming data into feelings, sounds, and other forms that create surprising analogies to already-understood principles. The result of such personal thinking is knowledge combined with sensation and emotion--feeling and understanding synthesized into complete awareness. We illustrate some of these modes of thinking with reference to recent breakthroughs in DNA-related areas and suggest ways in which the use of "tools for thinking" can increase the probability of making further discoveries in the biomedical sciences. PMID- 11860298 TI - Utilizing genomic DNA purified from clotted blood samples for single nucleotide polymorphism genotyping. AB - CONTEXT: Linking single nucleotide polymorphisms to disease etiology is expected to result in a substantial increase in the number of genetic tests available and performed at clinical laboratories. Whole blood serves as the most common DNA source for these tests. Because the number of blood samples rises with the number of genetic tests performed, alternative DNA sources will become important. One such alternative source is clotted blood, a by-product of serum extraction. Efficiently using an already procured blood sample would limit the overall number of samples processed by clinical laboratories. OBJECTIVE: To determine if DNA purified from clotted blood can be effectively used for single nucleotide polymorphism genotyping. DESIGN: DNA was purified from the clotted blood of 15 donors. Single nucleotide polymorphism genotyping for the methylenetetrahydrofolate reductase and factor V Leiden mutations was performed with each DNA sample by 2 independent methods. RESULTS: High-quality DNA was obtained from each of the 15 individual clotted blood samples as demonstrated by UV spectrophotometric analysis, gel electrophoresis, and polymerase chain reaction amplification. The DNA was used successfully to obtain genotype data from both the methylenetetrahydrofolate reductase and factor V single nucleotide polymorphism assays for all samples tested. CONCLUSIONS: Clotted blood is a clinically abundant sample type that can be used as a source of high-quality DNA for single nucleotide polymorphism genotyping. PMID- 11860299 TI - Mitochondrial DNA--related mitochondrial dysfunction in neurodegenerative diseases. AB - Mitochondrial dysfunction occurs in several late-onset neurodegenerative diseases. Determining its origin and significance may provide insight into the pathogeneses of these disorders. Regarding origin, one hypothesis proposes mitochondrial dysfunction is driven by mitochondrial DNA (mtDNA) aberration. This hypothesis is primarily supported by data from studies of cytoplasmic hybrid (cybrid) cell lines, which facilitate the study of mitochondrial genotype phenotype relationships. In cybrid cell lines in which mtDNA from persons with certain neurodegenerative diseases is assessed, mitochondrial physiology is altered in ways that are potentially relevant to programmed cell death pathways. Connecting mtDNA-related mitochondrial dysfunction with programmed cell death underscores the crucial if not central role for these organelles in neurodegenerative pathophysiology. This review discusses the cybrid technique and summarizes cybrid data implicating mtDNA-related mitochondrial dysfunction in certain neurodegenerative diseases. PMID- 11860300 TI - Human lymphocyte antigen molecular typing: how to identify the 1250+ alleles out there. AB - The human lymphocyte antigen (HLA) typing community was one of the early groups to adopt molecular testing. This action was borne out of the need to identify the many alleles of the highly polymorphic HLA system. Early paradigms used restriction fragment length polymorphism regimes, but the polymerase chain reaction method of amplification quickly replaced that less-than-discriminating choice. Methods currently in use for HLA typing, with commercial kits available, are sequence-specific oligonucleotide probe (both dot blot and the reverse blot dot), sequence-specific primer amplification, restriction fragment length polymorphism of amplified products, double-stranded sequence conformation polymorphism (with and without reference strand), sequence-based typing, and microarray technologies. More than 1250 alleles are recognized by the World Health Organization and meet their criteria for assignment. These alleles can be identified by molecular methods and represent alleles present at class I and class II loci of the HLA complex. On occasion, ambiguous results still persist, even with the best molecular typing methods. Therefore, it is clear to the HLA typing community that a combination of the above methods may be needed to allow true discrimination of the possible alleles an individual carries in their genetic makeup. It is also clear that a typing laboratory may need to resort to nonmolecular serology to understand the significance and impact of the type generated by the HLA molecular typing laboratory. PMID- 11860301 TI - Molecular testing in the diagnosis and management of hepatitis C virus infection. AB - OBJECTIVES: To review hepatitis C virus (HCV), describe the types of molecular based tests available for the diagnosis and management of HCV infection, and discuss the appropriate utilization of these tests. DATA SOURCES: Current information is presented from the published literature, as well as new information where available. STUDY SELECTION: A major cause of posttransfusion and community-acquired non-A, non-B hepatitis worldwide is HCV. Approximately 4 million people in the United States are infected with HCV, resulting in 8000 to 10,000 deaths annually. Because HCV is not readily cultured, in vitro molecular based tests have been developed for use in the diagnosis and treatment of HCV infected patients. Molecular tests include qualitative and quantitative nucleic acid amplification tests, branched DNA tests, and HCV genotyping assays. Qualitative HCV nucleic acid amplification tests are used routinely in association with serologic tests to help make a diagnosis of infection with HCV. Quantitative HCV testing and genotyping methods have been found to be valuable tools in the treatment of infected patients. A patient's pretreatment HCV viral load and the rate of virus decline during therapy have been shown to correlate with the likelihood of long-term response to antiviral therapy. Information pertaining to the genotype of HCV infecting patients has been shown to be helpful in making recommendations regarding treatment. Certain genotypes of HCV are much more responsive to therapy, allowing a shorter course of treatment. CONCLUSIONS: Molecular tests are valuable tools for use in the diagnosis and treatment of patients infected with HCV. PMID- 11860302 TI - The microbiology laboratory's role in response to bioterrorism. AB - CONTEXT: Bioterrorism has existed since before the 14th century; however, the specter of such an attack is much greater today than ever before. Technical expertise in microbiology and molecular testing, combined with the rapidity of worldwide air travel, has ensured that no geographic area would be untouched in a widespread attack. Clinical microbiology laboratories will play a pivotal role in the detection of attacks involving weapons of mass destruction. OBJECTIVE: To identify and discuss the microorganisms most likely to be used as agents of bioterrorism. DATA SOURCES: Data were obtained from literature searches from 1997 through June 2001 using the subject headings of bioterrorism, biological weapons, biological warfare, anthrax, brucellosis, tularemia, smallpox, plague, and botulism. In addition, information was obtained from publications of the Center for Civilian Studies, Johns Hopkins University, the Centers for Disease Control and Prevention, American Society for Microbiology, and the United States Army Medical Research Institute of Infectious Diseases. DATA EXTRACTION AND SYNTHESIS: Findings obtained from these studies and publications were analyzed for the most likely microorganisms that would be involved in a bioterrorist attack and the most efficient means by which they could be identified. In all instances, the guidelines from the Centers for Disease Control and Prevention for Level A laboratories were observed. CONCLUSIONS: The most likely microorganisms to be utilized as biological weapons include Bacillus anthracis (anthrax), Brucella species (brucellosis), Clostridium botulinum (botulism), Francisella tularensis (tularemia), Yersinia pestis (plague), and variola major (smallpox). While knowledge of the potential of these microorganisms is critical, clinical microbiologists and medical technologists possess the basic tools to rule out the suspected pathogens or to refer these isolates to public health laboratories for identification and susceptibility testing. PMID- 11860303 TI - Genetic polymorphisms associated with venous and arterial thrombosis: an overview. AB - OBJECTIVE: To provide an overview of genetic polymorphisms associated with thrombotic cardiovascular disease. DATA SOURCES: A literature search using the National Library of Medicine database. STUDY SELECTION: The literature on genetic polymorphisms associated with venous and arterial thrombosis was reviewed. DATA EXTRACTION: Based on the literature review, the clinical significance of polymorphisms in various coagulation proteins was assessed and a summary was developed. CONCLUSIONS: Thrombosis is a multifactorial disorder, with both congenital and acquired risk factors. It is now clear that there are many genetic abnormalities that impart an increased risk for thrombophilia, and the presence of more than 1 abnormality results in a further increased risk of thrombosis. In hemostasis, there is a balance between procoagulant factors and natural anticoagulant proteins. The first genetic thrombotic disorders described were deficiencies of the natural anticoagulants, such as antithrombin, protein C, and protein S, but these abnormalities are rare and are caused by many different mutations. More recently, single polymorphisms that are relatively common in the general population have been described in procoagulant factors, such as factor V and prothrombin, which impart an increased risk for venous thrombosis. As more scrutiny is placed on the hemostatic system, further polymorphisms have come to light. The current challenge is to elucidate the relationship between these new polymorphisms and either venous or arterial thrombotic cardiovascular disease. PMID- 11860304 TI - Influence of platelet collagen receptor polymorphisms on risk for arterial thrombosis. AB - CONTEXT: Collagens are major components of the vascular subendothelium, and the interaction of platelets with collagens initiates normal hemostasis or pathologic arteriothrombosis. Genetic factors that affect the interaction of platelets with collagens could represent risk factors for either arteriothrombosis or excessive hemorrhage. In this regard, we first found that platelet levels of one of the major platelet collagen receptors, integrin alpha(2)beta(1), vary up to 10-fold in normal healthy individuals and that the higher-level phenotype is associated with allele 1 (807T) of the integrin alpha(2) gene. More recently, we found that there is roughly a fivefold range in platelet glycoprotein VI content among normal individuals, which may also influence risk for thromboembolism. OBJECTIVE: To determine if genetic polymorphisms of platelet glycoproteins involved in collagen-related function are associated with higher risk for thrombotic disorders, such as coronary heart disease, myocardial infarction, or stroke. METHODS: We examined the genetic mechanisms responsible for variation in expression levels of the collagen receptor integrin alpha(2)beta(1) and the potential influence of this variation on risk for thrombotic diseases. RESULTS: We found that patients with arteriothrombotic diseases have a higher frequency of alpha(2) allele 1 (associated with higher levels of platelet integrin alpha(2)beta(1)). We further found that platelet glycoprotein VI content directly correlates with platelet prothrombinase activity, suggesting that a higher phenotype of platelet glycoprotein VI also may contribute to increased risk of arteriothrombotic diseases. CONCLUSION: Genetic polymorphisms that influence the level or function of platelet collagen receptors need to be seriously considered as genetic risk factors for arteriothrombotic diseases. PMID- 11860305 TI - Trends in genomic analysis of the cardiovascular system. AB - OBJECTIVE: To evaluate the opportunities afforded cardiovascular medicine by the comprehensive and integrative approaches of genomics in cellular physiology. We present a meta-analysis of recently reported results obtained by means of high throughput technologies (complementary DNA and oligonucleotide arrays, serial analysis of gene expression [SAGE]), as well as more traditional molecular biology approaches (real-time polymerase chain reaction, differential display, and others). DATA SOURCES: Newly published articles identified on PubMed and additional data provided by authors on-line (where available). CONCLUSIONS: The impact of genomic analysis on cardiovascular research is already visible. New genes of cardiovascular interest have been discovered, while a number of known genes have been found to be changed in unexpected contexts. The patterns in the variation of expression of many genes correlate well with the models currently used to explain the pathogenesis of cardiovascular diseases. Much more work has yet to be done, however, for the full exploitation of the immense informative potential still dormant in the genomic technologies. PMID- 11860306 TI - Clinical trials in vascular gene therapy: the missing piece of the puzzle. AB - OBJECTIVE: Vascular gene transfer provides unique opportunities for novel therapies for cardiovascular disease. To define the limitations and expectations of translating preclinical strategies to clinical therapy, the kinetics and extent of vascular transgene expression must be determined. DATA SOURCES: Published clinical and preclinical studies of vascular gene transfer. DATA EXTRACTION: Kinetics and extent of vascular transgene expression. DATA SYNTHESIS: In spite of ongoing clinical studies in vascular gene transfer, data regarding the kinetics and extent of transgene expression are very limited. CONCLUSIONS: For the efficient translation of preclinical to clinical vascular gene transfer studies, the kinetics and extent of transgene expression should be determined. Novel imaging technologies and unique clinical scenarios for obtaining tissue should be considered. PMID- 11860307 TI - Prospects for microtechnology and nanotechnology in bioengineering of replacement microvessels. AB - CONTEXT: Due to its anticipated curative potential, therapeutic angiogenesis recently became a major preoccupation for the biomedical research community. Most of the related work reported to date employs either biochemical or genetic tools. OBJECTIVE: To identify opportunities for application of the current developments in microtechnology and nanotechnology to the field of therapeutic angiogenesis. DATA SOURCES: Survey of recent English-language literature on microvascular tissue engineering in the context of therapeutic angiogenesis. We include our results regarding the role played by microtopographical cues in the progression of angiogenesis, such as those produced during processing of the extracellular matrix by chronic inflammatory cells. CONCLUSION: While notable accomplishments have been identified in the field of tissue engineering of larger vessels, reports on purposeful assembly of microvascular structures with the ability to be transferred in vivo by implantation are still scarce. Under these circumstances, we suggest the development of a new class of implantable biomedical microdevices, that is, "angiogenesis assist devices" (or "angiochips"), and we indicate some of their conceivable applications. PMID- 11860308 TI - Morphologic spectrum of estrogen receptor-negative breast carcinoma. AB - CONTEXT: Estrogen receptor (ER)-negative breast carcinomas are a heterogeneous group of breast cancers that are generally thought to be aggressive. OBJECTIVE: To determine the morphologic and immunohistochemical spectrum of a consecutive series of ER-negative breast carcinomas, in an attempt to understand the pathogenesis and behavior of these lesions. DESIGN: Seventy-four consecutive cases of ER-negative invasive carcinomas were studied. Hematoxylin-eosin-stained sections were reviewed, and new sections were stained for c-erbB-2, p53, vimentin, and androgen and prolactin receptors. The findings were correlated with the axillary lymph node status as a measure of tumor aggressiveness. SETTING: The histopathology department of a tertiary referral teaching hospital. RESULTS: The tumors included 50 (68%) invasive ductal carcinomas, 21 (28%) medullary/atypical medullary carcinomas, and 1 each of invasive lobular, apocrine, and papillary carcinoma. Some of the invasive ductal cases had distinctive features that are described in this report. Maximum tumor diameter varied between 5 and 100 mm. Sixty tumors (81%) were grade 3, 13 (18%) were grade 2, and 1 (1%) was grade 1. Of the 60 cases in which the axillary node status was known, 34 (57%) had metastases, and 26 did not. Tumors associated with positive nodes were significantly larger than those associated with negative nodes (37.2 vs 17.8 mm, P <.001). A higher percentage of node-negative tumors were c-erbB-2 positive (42% vs 21%, P <.05). There were no differences between the 2 groups with regard to histologic type, tumor grade, or the expression of p53, vimentin, or androgen or prolactin receptors. CONCLUSIONS: Many ER-negative breast carcinomas have distinctive microscopic features. Not all ER-negative tumors are aggressive, as judged by the absence of lymph node metastases in 43% of cases in this series. Tumor size is the most important indicator for the likelihood of the presence of lymph node metastases. The wide range of tumor sizes encountered in this series suggests that the ER status of a tumor is determined early in its natural history and supports the existence of 2 separate pathways for the development of ER negative and ER-positive breast carcinomas. PMID- 11860309 TI - Assessment of Epstein-Barr virus association with pediatric non-hodgkin lymphoma in immunocompetent and in immunocompromised patients in Argentina. AB - CONTEXT: Epstein-Barr virus (EBV) has been classically associated with 3 malignancies, Burkitt lymphoma, B-cell lymphoproliferative syndromes, and nasopharyngeal carcinoma, and more recently with Hodgkin disease, T-cell lymphomas, and gastric and breast carcinomas, as well as with leiomyosarcoma and leiomyoma associated with immunosuppression. OBJECTIVE: To compare EBV expression in Argentine tumor samples with those reported elsewhere, we analyzed EBV expression in an Argentine pediatric population with non-Hodgkin lymphoma and correlated these results with clinical course and outcome. METHODS: We studied EBV presence by latent membrane protein-1 protein labeling by immunohistochemistry, by in situ hybridization, and by polymerase chain reaction for Epstein-Barr-encoded RNAs (EBERs) in formalin-fixed and paraffin-embedded non Hodgkin lymphoma tissue samples (collected retrospectively) from 32 pediatric patients at Ricardo Gutierrez Children's Hospital from 1993 to 2000. RESULTS: Eight out of the 32 (25%) non-Hodgkin lymphoma cases showed latent membrane protein-1 and EBERs by in situ hybridization positive staining in tumor cells. Among EBERs and latent membrane protein-1-positive cases, there were 5 immunocompromised patients, with either human immunodeficiency virus infection or primary immunodeficiency. The EBERs in situ hybridization results were confirmed by EBERs polymerase chain reaction in good-quality DNA from 11 samples, with 3 proving positive and 8 negative. CONCLUSIONS: The association of EBV with non Hodgkin lymphoma in the Argentine pediatric population was low (25%), and this figure rose to 100% when only the immunocompromised patients subgroup was considered, confirming that the virus is probably a cofactor in the lymphomagenesis of some but not all pediatric non-Hodgkin lymphoma. So far, no differences in clinical outcome are discernible between EBV-positive and EBV negative non-Hodgkin lymphoma patients. PMID- 11860310 TI - Changes in automated complete blood cell count and differential leukocyte count results induced by storage of blood at room temperature. AB - OBJECTIVE: To delineate changes that occur in various parameters of automated complete blood cell count (CBC) and differential leukocyte count (differential) on prolonged storage of blood at room temperature. DESIGN: A CBC and an automated differential were performed on the Coulter Gen.S on 40 K(3) (tripotassium ethylenediamine-tetraacetate) EDTA-anticoagulated blood specimens once daily, specimen volume permitting, for 3 to 7 days. Specimens were kept at room temperature throughout the study. The results were tabulated using a personal computer with Excel software. Percent change or absolute difference from the initial value for each parameter for each subsequent day of the study period was calculated. RESULTS: Among the CBC parameters, hemoglobin, red blood cell count, and mean corpuscular hemoglobin were stable for the duration of the study (7 days), white blood cell count was stable for at least 3 days (up to 7 days, if the count was within or above the normal range), and platelet count was stable for at least 4 days (up to 7 days, if the count was within or above the normal range). The mean corpuscular volume, mean platelet volume, hematocrit, and red blood cell distribution width each increased, and the mean corpuscular hemoglobin concentration decreased from day 2 onward. Among the differential parameters, the relative percentages and absolute numbers of neutrophils, lymphocytes, and eosinophils tended to increase, whereas those of monocytes trended downward over time. Limited data on basophils did not reveal an appreciable change. CONCLUSION: Blood specimens stored at room temperature for more than 1 day (up to 3 days or possibly longer) were found to be acceptable with some limitations for CBC but not for the differential. PMID- 11860311 TI - Cosleeping and sudden unexpected death in infancy. AB - CONTEXT: The practice of infants cosleeping with adults has long been the subject of controversy. Autopsy findings in cases of sudden infant death syndrome (SIDS) are usually indistinguishable from those found with unintentional or intentional suffocation, and the determination of the cause of death in cases of sudden unexpected death in infancy is often based on investigative findings and the exclusion of natural or traumatic causes. OBJECTIVE: To further elucidate the risk of cosleeping. METHODS: We reviewed 58 cases of sudden unexpected infant deaths. Cases were excluded if there was any significant medical history or evidence of trauma or abuse. RESULTS: Twenty-seven of the infants were cosleeping. Eleven of these cases had been previously diagnosed as SIDS, and in 7 cases parental intoxication was documented. CONCLUSION: Our findings support recent studies that suggest that cosleeping or placing an infant in an adult bed is a potentially dangerous practice. The frequency of cosleeping among cases diagnosed as SIDS in our study suggests that some of these deaths may actually be caused by mechanical asphyxia due to unintentional suffocation by the cosleeping adult and/or compressible bedding materials. PMID- 11860312 TI - Group for research in pathology education online resources to facilitate pathology instruction. AB - BACKGROUND: The Group for Research in Pathology Education (GRIPE) is an organization of pathology educators whose purpose is to promote and facilitate excellence in pathology education. One important function of GRIPE is the maintenance of image and multiple-choice test question data banks. These resources have recently been made available online via the GRIPE Digital Library Web site. The purpose of the GRIPE Digital Library project was to develop an online searchable database that would facilitate access to the GRIPE resources for pathology education. DESIGN: The GRIPE image bank--containing approximately 3000 peer-reviewed gross and microscopic pathologic images along with textual descriptions--was linked with the GRIPE test question bank using Gossamer Thread's DBMan Web database management program. The search and display templates create a functional user interface that integrates images, image descriptions, and test questions into a single online digital library. Using any Web browser, faculty can access the GRIPE Digital Library and search for images and/or test items that can be used in teaching. RESULTS: In the first 18 months (February 2000 through July 2001), users at 40 GRIPE member institutions signed up and used the GRIPE Digital Library to perform more than 6000 individual searches and view more than 37500 images. These digital images were used to produce lectures and laboratory modules that were posted on Web pages and made available to students remotely. CONCLUSIONS: The GRIPE Digital Library provides a unique resource that can facilitate development of educational materials for pathology instruction and helps to fulfill the educational mission of GRIPE. PMID- 11860313 TI - Extranodal posttransplant plasmacytic hyperplasia with subsequent posttransplant plasmacytic malignancy: six-year interval case report and review of the literature. AB - Posttransplant lymphoproliferative disorders (PTLDs) represent a morphologic, immunophenotypic, and genotypic spectrum of disease. Most recently, Knowles et al divided PTLDs into 3 distinct categories: (1) plasmacytic hyperplasia, (2) polymorphic B-cell hyperplasia and polymorphic B-cell lymphoma, and (3) immunoblastic lymphoma and multiple myeloma. Although one form of PTLD may progress to another form, only 1 previous case has been reported in which multiple myeloma developed 14 months after an original diagnosis of plasmacytic hyperplasia. The type of solid organ transplant was not specified in that case. We report a post--cardiac transplant plasmacytic hyperplasia developing 7 years posttransplant. Six years subsequent to the plasmacytic hyperplasia, the patient developed a posttransplant plasmacytic malignancy, supported by morphology, flow cytometric immunophenotyping, and genotypic studies. Since we have no data to support disseminated bony disease or an abnormal serum protein, we have not used the term "multiple myeloma" for this case. PMID- 11860315 TI - Postmortem diagnosis of "occult" Klinefelter syndrome in a patient with chronic renal disease and liver cirrhosis. AB - This report describes a patient not suspected of having Klinefelter syndrome during life but diagnosed with it following postmortem examination using fluorescent in situ hybridization (FISH) for sex chromosomes and hormone serum analysis. A 49-year-old Japanese man had a history of nephrosis, heavy alcohol consumption, diabetes mellitus, and liver cirrhosis and had been undergoing dialysis for 10 years. He died of ruptured esophageal varices. Autopsy revealed hypogonadism, suggesting Klinefelter syndrome. This was confirmed by FISH, which showed a mosaic 46XY, 47XXY karyotype, and by serum analysis, which revealed high luteinizing hormone and follicle-stimulating hormone and low testosterone levels. Autopsy also revealed a nodular, bilateral, testicular Leydig cell hyperplasia. This report illustrates the value of postmortem laboratory investigations, particularly FISH for sex chromosomes and serum hormone analysis, for the demonstration of clinically uncertain or "occult" Klinefelter syndrome. PMID- 11860314 TI - Cavernous hemangioma of spermatic cord: report of a case with immunohistochemical study. AB - We present a case of spermatic cord cavernous hemangioma. A 32-year-old man presented with a circumscribed, painless mass in the left side of the spermatic cord. An orchiectomy of the left testicle was performed. A 3 x 3 x 2.5-cm mass was present in the spermatic cord area. Histologic examination and immunohistochemical study showed a benign vascular tumor composed of vascular spaces of varying size. Although cavernous hemangioma can occur in any location, the spermatic cord is an extremely rare site, and, to our knowledge, only a few cases have been previously reported. PMID- 11860316 TI - Mesenteric paraganglioma: a case report and review of the literature. AB - Approximately 5% to 10% of paragangliomas occur in extra-adrenal sites, which can extend from the upper cervical region to the pelvis, parallel to the autonomic nervous system. This distribution corresponds to the embryologic development of the paraganglia from neural crest cells. Rarely, extra-adrenal paragangliomas can also occur aberrantly outside this distribution. We report such a case of extra adrenal paraganglioma occurring in the anterior mesentery in a 76-year-old man. Two case reports exist in the literature describing extra-adrenal paragangliomas in the posterior mesentery. Normal paraganglionic tissue has been described at the roots of the superior and inferior mesenteric arteries, theoretically explaining the origin of the posterior mesenteric paragangliomas. Our case can best be attributed to the ventral migration of paraganglionic tissue through these vessels to reach the anterior mesentery, where they could potentially give rise to paragangliomas in this site. PMID- 11860317 TI - Multiple myeloma in association with sarcoidosis. AB - The association of sarcoidosis with Hodgkin disease and non-Hodgkin lymphoma is well known. However, multiple myeloma also can occur rarely in association with sarcoidosis. We describe a patient with sarcoidosis who subsequently developed multiple myeloma. The patient was a 49-year-old woman with a 4-year history of severe, chronic, active sarcoidosis involving her lungs, lymph nodes, eyes, and bone marrow. During the initial clinical workup, a serum monoclonal paraprotein was detected and bone marrow examination revealed a slight increase in plasma cells (4%), in addition to noncaseating granulomas. Thus, the diagnoses of monoclonal gammopathy of undetermined significance and sarcoidosis were established simultaneously. She sought medical attention for her current illness when she developed low back pain and weakness of her lower extremities. Serum protein electrophoresis and immunofixation revealed a monoclonal paraprotein, immunoglobulin (Ig) G kappa type, and quantification revealed an IgG level of 46.67 g/L (normal, 5.88--15.73 g/L). Bone marrow aspiration and biopsy revealed multiple myeloma and sarcoidosis. Including this patient, 11 cases of sarcoidosis and multiple myeloma have been reported to date, including 3 patients with monoclonal gammopathy of undetermined significance preceding the onset of multiple myeloma. In this case, as in most of the cases reported previously, sarcoidosis preceded the development of multiple myeloma. PMID- 11860318 TI - Malignant intrahepatic biliary papillomatosis associated with viral C cirrhosis. AB - Biliary papillomatosis is a rare entity characterized by multiple papillary adenomas involving extensive areas of the biliary tract with a great potential for recurrence and malignant transformation. It has been reported in association with Caroli disease and a choledochal cyst. We report herein a case of malignant intrahepatic biliary papillomatosis associated with cirrhosis secondary to hepatitis C. To the best of our knowledge, this is the first report of this association. PMID- 11860319 TI - Malakoplakia of liver diagnosed by a needle core biopsy: a case report and review of the literature. AB - Although malakoplakia of the genitourinary tract and colon is reported frequently in the literature, malakoplakia that occurs primarily in the liver is rare, and only 4 cases have been described thus far. To our knowledge, this is the first case of malakoplakia of the liver diagnosed by a needle core biopsy. This case occurred in a 19-year-old man with small bowel ileus following Klebsiella pneumonia. PMID- 11860320 TI - Pathologic quiz case: patient with systematic sclerosis and anemia. PMID- 11860321 TI - Pathologic quiz case: a papillary ovarian tumor in a 4-year-old-girl. PMID- 11860322 TI - Pathologic quiz case: a 3-year-old boy with new-onset partial seizures. PMID- 11860323 TI - Enterocolonic Mycobacterium avium-intracellulare. PMID- 11860324 TI - Kala azar associated with malaria. PMID- 11860325 TI - Attention and driving in traumatic brain injury: a question of coping with time pressure. AB - BACKGROUND: Diffuse and focal traumatic brain injury (TBI) can result in perceptual, cognitive, and motor dysfunction possibly leading to activity limitations in driving. Characteristic dysfunctions for severe diffuse TBI are confronted with function requirements derived from the hierarchical task analysis of driving skill. OBJECTIVE: Specifically, we focus on slow information processing, divided attention, and the development of procedural knowledge. Also the effects of a combination of diffuse and focal dysfunctions, specifically homonymous hemianopia and the dysexecutive syndrome, are discussed. Finally, we turn to problems and challenges with regard to assessment and rehabilitation methods in the areas of driving and fitness to drive. PMID- 11860326 TI - Useful field of view after traumatic brain injury. AB - BACKGROUND: Traumatic brain injury (TBI) survivors often have sensory and cognitive impairments that may interfere with driving ability. The Useful Field of View (UFOV) is a measure of visual information processing that is a good predictor of vehicle crash risk in older adults. OBJECTIVE: The objective of this study was to explore the possibility that UFOV is compromised after TBI. DESIGN: UFOV performance of 23 TBI survivors and 18 young adults without neurological impairment were compared. CONCLUSION: TBI survivors had higher UFOV scores than young adults, which indicated a greater functional loss of peripheral vision. The results suggest that the UFOV may be a valuable instrument for assessing driving readiness in TBI survivors. PMID- 11860327 TI - Divided attention and driving: a pilot study using virtual reality technology. AB - BACKGROUND: Virtual reality (VR) was used to investigate the influence of divided attention (simple versus complex) on driving performance (speed control). DESIGN: Three individuals with traumatic brain injury (TBI) and three healthy controls (HC), matched for age, education, and gender, were examined. RESULTS: Preliminary results revealed no differences on driving speed between TBI and HC. In contrast, TBI subjects demonstrated a greater number of errors on a secondary task performed while driving. CONCLUSION: The findings suggest that VR may provide an innovative medium for direct evaluation of basic cognitive functions (ie, divided attention) and its impact on everyday tasks (ie, driving) not previously available through traditional neuropsychological measures. PMID- 11860328 TI - Driving behaviors following brain injury: self-report and motor vehicle records. AB - BACKGROUND: This study examined both objective and subjective measures of driving behaviors occurring in the past 5 years for 47 individuals with traumatic brain injury (TBI) and 22 healthy controls (HC), matched for age, gender, education, and years of driving experience. RESULTS: Overall, subtle descriptive differences in driving characteristics were observed between the two groups. However, comparison of self-reported and documented reports of aberrant driving behaviors did not reveal a significantly greater number of accidents or violations among TBI participants compared with HC drivers. The results suggest that individuals with TBI, who successfully complete a driving evaluation program, are able to reintegrate into the driving community with minimal difficulty. PMID- 11860329 TI - Driving assessment issues for practicing clinicians. AB - BACKGROUND: Practical issues facing rehabilitation clinicians working with survivors of brain injuries who wish to return to motor vehicle operations are considerable, and this challenge will likely only increase in the future. Although unpleasant, clinicians cannot avoid the legal and ethical issues associated with this aspect of rehabilitation. Clinicians also face numerous problems in terms of a variety of assessment and treatment strategies, most with limited demonstrated ecological validity, and few, if any, clear guidelines for rehabilitation of the driver. Clinicians may not have access to "driving tests," simulators and computer programs, specialized personnel, or adaptive driving programs. CONCLUSION: Practical clinical issues such as these are reviewed, along with suggestions for a number of effective clinical approaches. PMID- 11860330 TI - Funding brain injury programs: the federal appropriations process. PMID- 11860331 TI - Assessment in traumatic brain injury: update on recent developments. PMID- 11860332 TI - The minimally conscious patient: when can life support be terminated? PMID- 11860335 TI - Random sequence libraries displayed on phage: identification of biologically important molecules. AB - Phage display has become a widely used tool for the identification of proteins or peptides with affinity for a variety of biomolecules. The versatility, simplicity and cost effectiveness of this application has pervaded a wide variety of research areas. Although not without its limitations, phage display has provided a convenient methodology for obtaining ligands to study the function, structure and diagnostic or therapeutic potential of various macromolecules. This review highlights some recent research employing this technology that serves to illustrate its utility in various research and clinical applications. PMID- 11860336 TI - Mimotopes of viral antigens and biologically important molecules as candidate vaccines and potential immunotherapeutics. AB - Antigen recognition by antibodies or ligand-receptor interactions involve small areas of the molecule named epitopes that are normally conformational in nature. The availability of combinatorial peptide libraries has provided a powerful tool for selecting novel sequences which mimic conformational epitopes (mimotopes) either structurally and/or immunologically. These mimotopes can be particularly useful in a number of situations, including: the development of vaccines against tumors, infectious diseases or allergic conditions; the design of molecules which can act as agonists or antagonists of various biologically-important molecules; and for the development of diagnostic assays. This article reviews the authors work on the application of combinatorial peptide libraries to identify mimotopes of protective B-cell epitopes from various pathogens, and the search for molecules able to block the biological activities of TNF-alpha, a cytokine which plays a key role in inflammation. PMID- 11860337 TI - Rapid determination of pharmacokinetic properties of new chemical entities: in vivo approaches. AB - There is a continuing need for increased throughput in the evaluation of new chemical entities in terms of their pharmacokinetic (PK) parameters as part of new drug discovery. This review summarizes various approaches that have been used to increase throughput in this area. The article divides the approaches into two areas: assay enhancement and sample reduction. PMID- 11860338 TI - A systematic SAR study of C10 modified paclitaxel analogues using a combinatorial approach. AB - A library with 63 paclitaxel analogues modified at the C10 position of paclitaxel has been prepared using parallel solution phase synthesis. Most of the C10 analogues were slightly less active than paclitaxel in the tubulin assembly assay and had reduced potency in the B16 melanoma and MCF-7 cell line cytotoxicity assays. These modifications at C10, however, did not lead to the total loss of activity, indicating that the C10 moiety of paclitaxel may not be directly involved in the drug-microtubule interactions, but could influence its binding affinity to P-glycoprotein. Approximately 50% of the analogues demonstrated better activity against the drug resistant cell line MCF7-ADR. However, the increase in activity was 10-fold at most. This result demonstrates that the cytotoxicity against this drug resistant cancer cell line is sensitive to structural changes at the C10 position of paclitaxel. It was also found that the presence of a nitrogen atom in the C10 substituent might play a role in the interaction of analogues with microtubules. PMID- 11860339 TI - Molecular diversity sample generation on the basis of quantum-mechanical computations and principal component analysis. AB - The present study introduces a new strategy of selection of a maximum diversity sample of n compounds from N available in a molecular database. This strategy can be useful in pharmacological screening, combinatorial chemistry or parallel synthesis planning. It consists of first describing the compounds by means of parameters derived from quantum mechanical computations (water solvation deltaG, benzene solvation deltaG, octanol solvation deltaG, dipolar moment), as well as standard molecular parameters such as solvent-accessible surface area and molecular weight. Solvation parameters are used because of the importance of this phenomenon in the pharmacological behaviour. Redundant information in the description of the compounds is eliminated by using principal components (PC) instead of the original descriptors. Based on the similarity between the N compounds in the PC space, they are classified into n groups by k-means cluster analysis. The compounds that are nearest to the centroid of each cluster constituted the maximum diversity sample. When practical difficulties exist for the use of one of the proposed compounds, another also close to the cluster centroid can substitute for it. This strategy has been tested in the selection of a sample of 50 amines from the 923 available in the Aldrich catalogue. The results have been contrasted with those obtained from an optimal, distance-based experimental design, resulting in an 86% of agreement between both approaches. An R(2)-like diversity coefficient has been used to assess the quality of the proposed solutions. PMID- 11860340 TI - Screening drugs for metabolic stability using pulsed ultrafiltration mass spectrometry. AB - A pulsed ultrafiltration-mass spectrometric screening method has been developed to evaluate the metabolic stability of drugs. Pooled human liver microsomes containing cytochrome P450 enzymes were trapped by an ultrafiltration membrane in a stirred flow-through chamber, and eight beta-blocker drugs including acebutolol, alprenolol, atenolol, metoprolol, oxprenolol, pindolol, propranolol, and timolol were flow-injected through the chamber along with the cofactor NADPH. The ultrafiltrate was collected, concentrated and analyzed by using liquid chromatography-tandem mass spectrometry (LC-MS-MS) in order to quantitate the unmetabolized fraction of each drug. The metabolic stability of each beta-blocker was determined based on the difference between the corresponding LC-MS-MS peak areas of an experimental incubation and a control without NADPH. A flow-through incubation method, pulsed ultrafiltration metabolic screening minimizes the potential for product feed back inhibition of cytochrome P450 enzymes. The importance of this phenomenon was illustrated by the observation that the metabolic stability of the set of beta-blocker drugs measured using pulsed ultrafiltration more closely resembled the in vivo stability than that determined using a conventional batch incubation with microsomes or an incubation with human hepatocytes. Since a mixture of compounds was analyzed, the relative metabolic stability of each compound could be assessed by comparison to the other compounds in the incubation. This approach might be particularly useful for the ranking of a directed library of drug leads with respect to metabolic stability and then the selection of lead compounds for further drug development. PMID- 11860341 TI - Evaluation of a micro volume pulsed ultrafiltration cell for screening ligands in non-covalent complexes. AB - A pulsed ultrafiltration cell with a 35 microL binding chamber was evaluated for its ability to screen ligands that formed non-covalent complexes with protein targets. The cell was tested with ligands to the targets of carbonic anhydrase and serum albumin. Non-covalent ligand binding to both of these targets was observed and bound ligands were eluted from the cell in less than five min. The cell was also demonstrated to effectively screen a methanolic fermentation broth extract spiked with a known inhibitor to carbonic anhydrase. In addition to detecting specific binding events, the pulsed ultrafiltration method was investigated for its ability to distinguish non-specific binding events. Using carbonic anhydrase with the zinc-binding site removed, it was found that non specific complexes observed when using electrospray ionization alone were not detected when using the pulsed ultrafiltration mass spectrometry method. PMID- 11860342 TI - Solid-phase and solution-phase parallel synthesis of tetrahydro-isoquinolines via Pictet-Spengler reaction. AB - An efficient parallel synthesis of 6,7-dimethoxytetrahydroisoquinolines is reported. The key reaction step is 3,4-dimethoxyphenethylimines reacting with acid chlorides to form an N-acyliminium ion intermediate, which undergoes Pictet Spengler condensation to give the desired products in >80% yield. Both solution phase and solid-phase synthesis of 6,7-dimethoxytetrahydroisoquinolines are described. PMID- 11860343 TI - Recognition of multiple substrate motifs by the c-ABL protein tyrosine kinase. AB - Using a combinatorial peptide library that is based on the one-bead one-peptide approach we identified 14 peptide substrates for the c-ABL protein tyrosine kinase, which define three distinct consensus sequence groups. This is distinct from many serine/threonine kinases, which often phosphorylate only one major consensus sequence. The three consensus sequences accurately predict phosphorylation sites in cellular ABL substrates proven to play a role in cell signaling. Our data suggest that protein tyrosine kinases have evolved to recognize multiple substrate motifs. PMID- 11860344 TI - Solid phase synthesis of structurally diverse tetra substituted pyrimidines for potential use in combinatorial chemistry. AB - A new pyrimidine based scaffold has been identified for generation of combinatorial libraries using solid phase technique. The utility of the scaffolds was demonstrated by synthesizing small libraries of 12 substituted pyrimidines (4a-4l). PMID- 11860345 TI - [Dmt(1)]DALDA is highly selective and potent at mu opioid receptors, but is not cross-tolerant with systemic morphine. AB - The clinical effectiveness of morphine is limited by several side effects, including the development of tolerance and dependence. Most of these side effects are believed to be mediated by central opioid receptors; therefore, hydrophilic opioids, which don't cross the blood-brain barrier, may have advantages over morphine in some clinical applications. We recently synthesized several analogues of DALDA (Tyr-D-Arg-Phe-Lys-NH2), a highly hydrophilic peptide derived from the endogenous opioid peptide dermorphin; all of them, particularly [Dmt(1)] DALDA (Dmt - 2',6'-dimethyl tyrosine), had high potency and selectivity at mu receptors, the target of morphine, in activity assays. Here we report the pharmacological characterization of [Dmt(1)] DALDA in the whole animal. [Dmt(1)]DALDA was 40 times more potent than morphine in inducing antinociception in mice when both drugs were given s.c., and 6-14 times more potent than DAMGO, a selective m agonist, when both drugs were given it. However, [Dmt(1)]DALDA showed poor cross-tolerance to morphine; thus chronic morphine treatment of animals increased the antinociceptive AD(50) of systemic [Dmt(1)]DALDA two fold or less, as compared to an 8-9-fold increase for morphine and a 4-5-fold increase for DAMGO. The antinociceptive activity of [Dmt(1)]DALDA (i.t) was blocked by CTAP, a selective mu antagonist, but not by TIPP psi, a selective delta antagonist, nor by nor-BNI, a selective kappa antagonist. [Dmt(1)]DALDA-induced antinociception was also blocked by naloxone methiodide, an antagonist that does not cross the blood-brain barrier, when agonist and antagonist were given i.t. or i.c.v., but not when they were given s.c. We conclude that [Dmt(1)] DALDA is a highly potent analgesic acting at mu receptors. Though it appears to penetrate the blood-brain barrier, it exhibits low cross-tolerance to morphine, suggesting that it may have advantages over the latter in certain clinical applications. PMID- 11860346 TI - The cellular electrophysiologic effect of a new amiodarone like antiarrhythmic drug GYKI 16638 in undiseased human ventricular muscle: comparison with sotalol and mexiletine. AB - The cellular electrophysiologic effect of GYKI 16638, a new antiarrhythmic compound was studied and compared with that of sotalol and mexiletine in undiseased human right ventricular muscle preparation by applying the conventional microelectrode technique. GYKI 16638 (5 microM), at stimulation cycle length of 1000 ms, lengthened action potential duration (APD(90)) from 338.9 +/- 28.6 ms to 385.4 +/- 24 ms (n = 9, p > 0.05). This APD lengthening effect, unlike that of sotalol (30 microM), was rate-independent. GYKI 16638, contrary to sotalol and like mexiletine (10 microM), exerted a use-dependent depression of the maximal rate of depolarization (V(max)) which amounted to 36.4 +/- 11.7% at cycle length of 400 ms (n = 5, p < 0.05) and was characterised with an offset kinetical time constant of 298.6 +/- 70.2 ms. It was concluded that GYKI 16638 in human ventricular muscle shows combined Class IB and Class III antiarrhythmic properties, resembling the electrophysiological manifestation seen after chronic amiodarone treatment. PMID- 11860347 TI - The influence of metabolism on the MAO-B inhibitory potency of selegiline. AB - (-)-Deprenyl (selegiline), a propargylamine derivative of methylamphetamine, is a potent, irreversible inhibitor of monoamine-oxidase type B (MAO-B). The MAO-B inhibitory effects of various doses (0.1-0.25-0.5 mg/kg) of (-)-deprenyl in rat brain and liver were compared, using either oral or subcutaneous drug administration. The intensity of the first pass metabolism of (-)-deprenyl was also estimated. The effect of pre-treatment with phenobarbitone (80 mg/kg i.p., daily for three days) or proadifen (SKF-525A, 50 mg/kg i.p., single dose) on the MAO-B inhibitory potency of (-)-deprenyl was also studied. The oral and subcutaneous administration of selegiline induced a significantly different degree of MAO-B enzyme inhibition in the rat brain, but not in the liver. The inhibitory potency of (-)-deprenyl on MAO-B activity was markedly influenced by pre-treatment of rats with an inducer (phenobarbitone), or an inhibitor (SKF 525A) of cytochrome P-450 mono-oxygenases in the liver. Our results suggest, that (-)-deprenyl is metabolised mainly in the liver by microsomal cytochrome P-450 dependent mono-oxygenases, and it has an intensive first-pass metabolism. The parent compound is responsible for the inhibition of MAO-B enzyme activity. PMID- 11860348 TI - New trends in the development of oral antidiabetic drugs. AB - A large number of oral antidiabetic agents are available today. This article provides a short review of the pharmacology and some clinical aspects of various oral antidiabetic drugs. It focuses mainly on the newest developing drugs (therapy of the near future) and on the most commonly used older groups for the common approach of every-day practice (sulphonylureas). The primary goal of this review is to compare the electrophysiological effects of glibenclamide in isolated normal and streptozotocin induced diabetic rats and alloxan induced rabbits ventricular preparations, while on the other hand to differentiate the hypoglycaemic sulphonylureas (0.1-1000 mmol/kg) according to their cardiovascular activity in healthy and diabetic animals. In vitro (1-100 micromol/l) as well as chronically treated (5 mg/kg for 10 weeks) glibenclamide prolonged the action potential duration in normal but failed to affect it in diabetic ventricular preparations. Our results suggest that the sensitivity to glibenclamide of K(ATP) channels in diabetic ventricular fibers is drastically decreased. The effects of different sulphonylureas (tolbutamide, glibenclamide, gliclazide, glimepiride) on ventricular ectopic beats as well as the duration of ventricular fibrillation induced by 10 min ischemia/50 min reperfusion in healthy and diabetic rats were compared. Tolbutamide and gliclazide dose-dependently enhanced both parameters both in healthy and diabetic groups. Glibenclamide in healthy rats increased, while in diabetic rats it decreased the arrhythmogenicity. Glimepiride depressed the arrhythmogenicity in both healthy and diabetic animals. Glimepiride proved to dose-dependently enhance the myocardial tissue flow in dog in contrast to glibenclamide. These results confirm that glimepiride has less cardiovascular actions than other sulphonylureas. From the newest oral antidiabetics this review tries to emphasize the most important basic pharmacological properties, mechanism of action, therapeutic use. PMID- 11860349 TI - Advances in nuclear oncology: microspheres for internal radionuclide therapy of liver tumours. AB - Liver metastases cause the majority of deaths from colorectal cancer, and response to chemotherapy and external radiotherapy is poor. An alternative is internal radionuclide therapy using (90)Y labeled microspheres. These microspheres are very stable and have a proven efficacy in the field of treatment of primary or metastatic hepatic cancer. Whilst these glass spheres showed encouraging results in patients, their high density is a serious drawback. Currently, other materials with lower densities and other radioisotopes are being investigated in order to optimize this promising new therapy. Three major radiolabeled microsphere materials, viz. glass, resin-based and polymer-based, are now available for therapy or are being tested in animals. In this review the preparation, stability and degradation of these spheres are discussed. PMID- 11860350 TI - Pathways of inflammatory activation in Alzheimer's disease: potential targets for disease modifying drugs. AB - In the human brain several cell types are capable of initiating and amplifying a brain specific inflammatory response involving the synthesis of cytokines, prostaglandins and oxygen free radicals. In Alzheimer's disease (AD), signs of an inflammatory activation of microglia and astroglia are present inside and outside amyloid deposits. Cell culture and animal models suggest an interactive relationship between inflammatory activation, reduced neuronal functioning and deposition of amyloid. The activation of inflammation-associated enzymes such as p38 mitogen-activated protein kinase (p38 MAPK) and cycloxygenase-2 (COX-2) is not restricted to glial cells but also found in neurons and may contribute to intraneuronal damage. Epidemiological studies have shown a reduced risk of AD among users of anti-inflammatory drugs. Therefore, anti-inflammatory drugs have become the focus of several new treatment strategies. Small clinical trials with non-steroidal anti-inflammatory drugs (NSAIDs) such as indomethacin and diclofenac showed a trend for a disease modifying effect, while clinical trials with steroids did not show a beneficial effect. NSAIDs may not only act on COX-2 but also inhibit COX-1 activity or activate peroxisome proliferator-activated receptor gamma (PPAR gamma). Among promising new strategies to reduce the inflammatory activation in the CNS interfering with intracellular pro inflammatory pathways has been shown to be effective in various cell culture and animal models. Inhibitors of p38MAPK and PPAR gamma agonists may be suitable agents to suppress inflammatory activation in AD. PMID- 11860351 TI - Non steroidal anti-inflammatory and anti-allergy agents. AB - Non steroidal anti-inflammatory drugs (NSAIDs) are among the most commonly used for inflammation therapy. The major drawback in using the NSAIDs is in their tendency to cause gastrointestinal toxicity. Since the roles of arachidonic acid (A.A) metabolites, as leukotrienes (Lts), prostaglandins (PGs) and thromboxanes (TXA(2)) as mediators of the inflammatory reaction were clarified, much effort has been made to develop inhibitors of the production of these chemical mediators as anti-inflammatory agents. These mediators also play important roles in some inflammatory or allergic diseases, acting either alone or in combination and inhibitors of 5-lipoxygenase (5-LOX) and/or cyclooxygenase isoforms 1,2 (COX-1,2) may be useful for the treatment of asthma, psoriasis and rheumatoid arthritis. Leukotrienes, the products of 5-LOX metabolism have been associated with immediate hypersensitivity reactions, anaphylaxis and asthma. In addition, active oxygen species (AOS) including superoxide anion (O(2)(-)), hydrogen peroxide, hydroxyl radical and ferric radical, mediate cell damage in a variety of pathophysiological conditions and are responsible for oxidative injury of enzymes, lipid membranes and DNA in living cells and tissues. Prostaglandins and leukotrienes in the arachidonate pathway linked with lipid peroxidation may amplify the oxidative damage. Nitric oxide (NO) plays also a role as an effector in inflammation, since PG and NO thought to be important in maintaining mucosal integrity. Dual or selective inhibitors, specific receptor antagonists, AOS scavengers, and NO donors have been under development for therapeutic application. Several classes of inhibitors have been identified and at least 12 major chemical series are known to affect PGs production directly. In this review, we account on our research work concerning NSAIDs combined with a reference of the recent literature. PMID- 11860352 TI - The 5-HT(3) and nACh ionotropic receptors: a perspective from the computational chemistry point of view. AB - Recent contributions applying Computational Chemistry to serotonin-3 and nicotinic acetylcholine ionotropic receptors are reviewed. These two receptors constitute a good example for the examination of the computational protocols that have been used to understand how they work. On the one hand, (5-HT(3)R) receptor mapping techniques have been mostly employed in its study and very few examples of receptor fitting have been appeared. On the other hand, (nAChR) has been studied mainly from the receptor fitting point of view, although many contributions using receptor mapping exist. In the first case, antagonists seems to be more important that agonists, so more works are devoted to them. In the second case, agonist development is the main issue. Although far for being complete, in either of the cases we have working pharmacophores as well as 3D models for their binding sites that are ready to be used as a starting guess to design potential drugs. It is noteworthy that the absence of crystallographic structure for these receptors has motivated the interest in their study, constituting an interesting and challenging field. Mutagenesis experiments have allowed the establishment of main amino acids that are essential in the receptor functioning and then, interaction models have been postulated. Although most of the models are speculative in nature, some of them have been proved to be valuable tools for drug design. This scientific field is already open and many areas are still unexplored. Computational tools for treating these issues exist in a wide variety and their rational application would produce the answers to the structure and functioning of these receptors. PMID- 11860353 TI - Pathogen inactivation in blood products. AB - A combination of the economic importance of blood as a resource and the advent of the AIDS epidemic has led to a requirement for improved pathogen screening techniques for donated blood. However, due to window periods where infective agents cannot be detected, the ability to disinfect of blood and its derivatives plasma, platelets and red blood cell concentrates has assumed great importance. Whereas conventional disinfection techniques such as solvent-detergent treatment or ultra-violet irradiation may be employed in plasma or protein concentrates, the collateral damage associated with such treatments disallows their use with cellular fractions. In many ways the pathogen selectivity required here is akin to standard antimicrobial chemotherapy but is complicated by the requirement for activity against the full range of microbes--viruses, bacteria, yeasts and protozoa--rather than simply antibacterial or antiviral etc. The recent problems due to prion agents in the blood supply mean that such agents should also be included in any proposed disinfecting regimen. Several new approaches to microbial disinfection have been proposed by academia and the "blood industrials" : targeted chemotherapy, photochemotherapy and photodynamic antimicrobial chemotherapy. PMID- 11860354 TI - Inhibitors of multidrug resistance to antitumor agents (MDR). AB - Multidrug resistance is one of the main obstacles in the chemotherapy of cancer. Its inhibition by combination of chemosensitizers with antitumor compounds is a very active field of research, since safe and potent reversal agents would be beneficial for clinical use. Most modulators act by binding to membrane transport proteins (specially P-gp and MRP) and inhibiting their drug-effluxing activity, or by indirect mechanisms related to phosphorylation of the transport proteins or expression of the mdr1 and mrp1 genes. The main body of the review focuses on the study of the known MDR modulators, which are classified according to their chemical structures. General structure-activity studies of this therapeutic group are hampered by the very heterogeneous chemical structure of the compounds, although some conclusions have been drawn from the study of homogeneous series of molecules. PMID- 11860355 TI - Progress towards the discovery of xanthine oxidase inhibitors. AB - Xanthine oxidase (XO) is a highly versatile flavoprotein enzyme, ubiquitous among species (from bacteria to human) and within the various tissues of mammals. The enzyme catalyses the oxidative hydroxylation of purine substrates at the molybdenum centre (the reductive half-reaction) and subsequent reduction of O(2) at the flavin centre with generation of reactive oxygen species (ROS), either superoxide anion radical or hydrogen peroxide (the oxidative half-reaction). Many diseases, or at least symptoms of diseases, arise from a deficiency or excess of a specific metabolite in the body. For an example of an excess of a particular metabolite that produces a disease state is the excess of uric acid which can led to gout. Inhibition of XO decreases the uric acid levels, and results in an antihyperuricemic effect. Allopurinol, first synthesised as a potential anticancer agent, is nowadays a clinically useful xanthine oxidase inhibitor used in the treatment of gout. There is overwhelming acceptance that xanthine oxidase serum levels are significantly increased in various pathological states like hepatitis, inflammation, ischemia-reperfusion, carcinogenesis and aging and that ROS generated in the enzymatic process are involved in oxidative damage. Thus, it may be possible that the inhibition of this enzymatic pathway would be beneficial. In this review the State of the Art will be presented, which includes a summary of the progress made over the past years in the knowledge of the structure and mechanism of the enzyme, associated pathological states, and in the efforts made towards the development of new xanthine oxidase inhibitors. PMID- 11860356 TI - Inhibitors of AP-1 and NF-kappa B mediated transcriptional activation: therapeutic potential in autoimmune diseases and structural diversity. AB - Cytokines and chemokines play a very important role in a number of inflammatory diseases. In activated T cells, transcription factors such as the activator protein-1 (AP-1) regulate IL-2 production and production of matrix metalloproteinases, the nuclear factor kappa B (NF-kappa B) is essential for the transcriptional regulation of the proinflammatory cytokines IL-1, IL-6, IL-8 and TNF alpha, and nuclear factor of activated T-cells (NFAT) is required for the transcriptional regulation of IL-2, IL-3, IL-4, IL-5, IL-8, IL-13, TNF alpha, and GM-CSF. During the last few years, several groups have developed inhibitors of AP 1, NF-kappa B or both, and NFAT. This review article presents the recent progress in the development of inhibitors for AP-1, NF-kappa B, and NFAT mediated transcriptional activation. PMID- 11860357 TI - Okadaic acid, useful tool for studying cellular processes. AB - One of the most interesting groups of substances of marine origin, from structural and pharmacological points of view are polyether toxins, which generally present a great diversity in size and potent biological activities. The subject of this review is limited to okadaic acid (OA). It was the first example of a group of polyether toxins produced by marine microalgae, which is responsible for the natural phenomena known as Diarrhetic Shellfish Poisoning, DSP red tides. These toxins are accumulated in the digestive glands of the shellfish with a disastrous effect upon the shellfish industry in many parts of the world. Thus, it has been demonstrated that OA is a highly selective inhibitor of protein phosphatases type 1 (PP1) and 2A (PP2A), subsequently that it causes dramatic increases in phosphorylation of numerous proteins as well as being a potent tumour promoter. For that reason, OA is an extremely useful tool for studying the cellular processes that are regulated by reversible phosphorylation of proteins as signal transduction, cell division and memory. PMID- 11860358 TI - Review of estrone sulfatase and its inhibitors--an important new target against hormone dependent breast cancer. AB - A high proportion (approximately 40%) of breast cancers are hormone dependent. The female hormones estradiol and androstenediol are believed to play a key role in the initiation and promotion of this disease. In the fight against hormone dependent breast cancers, extensive research has been undertaken to produce compounds which are potent inhibitors against the cytochrome P-450 enzyme aromatase (AR), which converts the C19 androgens to the C18 estrogens. However, the administration of AR inhibitors alone has failed to produce the expected decrease in plasma levels of estrone. The major impetus to the development of steroid sulfatase inhibitors has therefore been the realisation that in order to improve therapeutic response for women with hormone-dependent breast cancer, not only must the AR enzyme be inhibited, but also the synthesis of estrogens via alternative routes. The steroid sulfatase enzyme regulates the formation of estrone (which can subsequently be converted to the potent estrogen estradiol) from estrone sulfate, a steroid conjugate present in high concentrations in tissue and blood in women with breast cancer. The sulfatase enzyme system also controls the formation of dehydroepiandrosterone (DHEA) from the DHEA-sulfate. This is important since DHEA can be converted to 5-androstene-3 beta,17 beta diol, which possesses estrogenic properties capable of stimulating the growth of breast cancer cells in vitro and in vivo. Considerable progress has been made in recent years in the development of a number of potent steroid/estrone sulfatase inhibitors, as such both steroidal and non-steroidal compounds have been considered and a number of highly potent inhibitors have been produced and evaluated against what is now considered a crucial enzyme in the fight against hormone dependent breast cancer. The review therefore considers the work that has been undertaken to date, as well as possible future development with respect to dual inhibitors of both estrone sulfatase and AR. PMID- 11860359 TI - Polycarboxylate fluorescent indicators as ion concentration probes in biological systems. AB - A large number of techniques have been applied to monitor the function of free metal ions in biological systems. Fluorescent ion probes have evolved into an extremely useful tool for contemporary experimentalists. Polycarboxylate indicators are widely used in the determination of metal ion concentrations, especially due to their cell membrane permeability. The design of these probes required detailed knowledge in related fields of medicine, biology, and chemistry, and their preparation demanded the expertise of organic synthesis. In this review, the basic rationale for the selection of particular chemical structures are analyzed, synthetic pathways leading to the desired structures are presented, often via a retrosynthetic approach, and properties and relative advantages of the use of these probes are described. References to specific applications are limited, given the large number of reviews on related subjects. Topics such as those related to dextran conjugates that are broad enough to be the subject of a different review are not included, and leakage resistance and near-membrane probes are mentioned briefly in separate sections, although these are chemically similar to typical polycarboxylate dyes. While reference to topics mentioned in related reviews is unavoidable, presentation of material in this review is from the point of view of a medicinal chemist rather than that of the many experts using these pioneering techniques. PMID- 11860360 TI - Identification of enzyme inhibitors using combinatorial libraries. AB - Potent enzyme inhibitors have long been recognized as powerful tools for assessing the physiological roles of enzymes and have led to the therapeutic drugs able to modulate their activities in vivo. However, to be valuable tools such inhibitors should be selective so that they do not interfere with other members of the particular enzyme family. Combinatorial chemistry has proven to be a novel approach for the identification of molecules with a desired selectivity profile from the libraries of several million compounds. In recent years it has been extensively used in conjunction with computational methods for the development of potent inhibitors of therapeutically interesting targets. This review describes the various structurally diverse enzyme inhibitors identified by screening combinatorial libraries of peptides and small organic molecules. PMID- 11860361 TI - DNA-binding of drugs used in medicinal therapies. AB - The interactions of various low-molecular weight substances with DNA are naturally relevant mechanisms in the cellular cycle and so also used in medicinal treatment. Depending on the particular drug structure, DNA-binding modes like groove-binding, intercalating and/or stacking, give rise to supramolecular assemblies of the polynucleotides, as well as influence the DNA-protein binding. In this review, we compare the underlying molecular structures, including general aspects of DNA sequences, with the benefit in medicinal treatment. While so far interest in this field had mainly been devoted to isolated nucleic acid/drug interactions, the present paper will focus on drug efficiencies generating and influencing supramolecular organizations and their complex sequence-dependent structure-activity codes. In particular, the attention will be directed to stereoelectronic relationships. Spatial enantioselective properties are discussed in details. As examples, the drug self-assemblies, as well as the influence of drugs on supramolecular DNA formations are described. A hypothetical connection between drug-influenced DNA-toroids and the formation of micronuclei in tissues will be interpreted. PMID- 11860362 TI - Endothelin receptor antagonists: structures, synthesis, selectivity and therapeutic applications. AB - Endothelin (ET) was discovered in 1988 and is the most potent vasoconstrictive peptide known to date. It exists in three isoforms (ET-1 to ET-3) and acts on two endothelin receptor subtypes, the endothelin-A (ET(A))-receptor and the endothelin-B (ET(B))-receptor. Endothelin receptor antagonists are novel therapeutics in clinical development for different cardiovascular, cerebrovascular, and renal diseases. Several different structural classes of endothelin receptor antagonists have been discovered within the last decade, starting from peptidic- and peptidomimetic structures to small organic molecules suitable as therapeutics for oral administration. Focussing on the small organic molecules, the different structural classes of ET-receptor antagonists are described with respect to synthesis, structure-activity-relationships, receptor subtype-selectivity profile, and where possible, intended therapeutic indications. PMID- 11860363 TI - Methionine in and out of proteins: targets for drug design. AB - The increasing need for new antibiotics to overcome rapidly developing resistance mechanisms observed in clinical isolates of Gram-positive and Gram-negative eubacteria has placed critical emphasis on the search for new antibacterial enzyme targets and the structural and mechanistic investigation of such targets. Among these potential targets, the enzymes responsible for integrating the amino acid methionine into proteins, along with its subsequent post-translational modification and repair, have emerged as promising candidates for the development of novel antibiotics. As well, there is increasing evidence for the importance of several of these enzymes in the development of anti-cancer, anti-parasitic, and anti-atherosclerotic drugs. Within the last three years, the crystal structures of all of these enzymes have been determined, which offers an unprecedented source of structural information for inhibitor design. The development of combinatorial chemistry and high throughput screening procedures has quickly provided several potent, specific inhibitors for a number of these enzymes, particularly the peptide deformylase, methionine aminopeptidase, and methionyl tRNA synthetase enzymes. This review critically analyzes the future potential for inhibition of enzymes in this pathway, allowing for a pragmatic view of the success of inhibitor developments and highlighting areas in which further investigations are warranted. PMID- 11860364 TI - The cell biology of parasitism in Trypanosoma brucei: insights and drug targets from genomic approaches? AB - The African trypanosome, Trypanosoma brucei exhibits a complex, digenetic life cycle that alternates between the tsetse fly vector and the mammalian host. The life cycle is characterised by a complex series of cell type differentiations and variations in metabolism. In addition the trypanosome exhibits a particular cell biology that has become adapted for its role as a parasite. This article places some of these areas in a frame-work that considers the role of cellular processes in parasitism. I rehearse some conclusions from recent studies and provide hyphotheses and suggestions for future work. Areas debated include: cell surface protein expression, cell differentiation, endomembrane trafficking and protein targeting, the cytoskeleton,flagellum functions in motility, attachment and plasma membrane differentiation, organelle specialisations, control of cell cycle, parasite/host, parasite/parasite and parasite/vector interactions. The review also focuses on the likely impact of the genome project and reverse genetics in providing greater insight to these cellular processes and how, if coordinated with some alan by scientists and funding agencies, this may provide novel targets for future drug development. PMID- 11860365 TI - Chemotherapy of human African trypanosomiasis. AB - Human African trypanosomiasis or sleeping sickness is resurgent [1,2]. The disease is caused by subspecies of the parasitic haemoflagellate, Trypanosoma brucei. Infection starts with the bite of an infected tsetse fly (Glossina spp.). Parasites move from the site of infection to the draining lymphatic vessels and blood stream. The parasites proliferate within the bloodstream and later invade other tissues including the central nervous system. Once they have established themselves within the CNS, a progressive breakdown of neurological function accompanies the disease. Coma precedes death during this late phase. Two forms of the disease are recognised, one caused by Trypanosoma brucei rhodesiense, endemic in Eastern and Southern Africa, in which parasites rapidly invade the CNS causing death within weeks if untreated. T. b. gambiense, originally described in West Africa, but also widespread in Central Africa, proliferates more slowly and can take several years before establishing a CNS-involved infection. Many countries are in the midst of epidemics caused by gambiense-type parasites. Four drugs have been licensed to treat the disease [3]; two of them, pentamidine and suramin, are used prior to CNS involvement. The arsenic-based drug, melarsoprol is used once parasites are established in the CNS. The fourth, eflornithine, is effective against late stage disease caused by T. b. gambiense, but is ineffective against T. b. rhodesiense. Another drug, nifurtimox is licensed for South American trypanosomiasis but also been used in trials against melarsoprol-refractory late sage disease. This review focuses on what is known about modes of action of current drugs and discusses targets for future drug development. PMID- 11860366 TI - Basic cell biology of Trypanosoma cruzi. AB - The paper reviews basic aspects of the biology of Trypanosoma cruzi emphasizing the following topics: (a) developmental stages of the life cycle in the vertebrate and invertebrate hosts; (b) the cytoskeleton of the protozoan, especially the sub-pellicular microtubules; (c) the flagellum, emphasizing its attachment to the protozoan body through specialized junctions; (d) the kinetoplast-mitochondrion complex, describing its structural organization and the replication of the kinetoplast-DNA; (e) the peroxisome (glycosome) and its role in the metabolism of the cell; (f) the acidocalcisome, describing its morphology, biochemistry and functional role; (g) the cytostome and the endocytic pathway; (h) the organization of the endoplasmic reticulum-Golgi complex; (i) the nucleus, describing its structural organization during interphase and division, and (j) basic aspects of the process of interaction of the parasite with host cells. PMID- 11860367 TI - Chemotherapy of Chagas disease. AB - In this article we review the current status of chemotherapeutic approaches for the specific treatment of Chagas disease or American Trypanosomiasis, as well as new rational approaches being developed as a consequence on the increased understanding of the biochemistry and physiology of its causative agent, the protozoan parasite Trypanosoma cruzi. Currently available drugs (nitrofurans and nitroimidazoles), developed empirically over three decades ago, are unsatisfactory due to frequent toxic side effects and limited efficacy, particularly in the prevalent chronic form of the disease. Furthermore, studies of their mechanism of action have shown that their antiparasitic activity is inextricably linked to mammalian host toxicity. Recent advances in this field include the demonstration that new triazole derivatives, with selective inhibitory activity on the parasite's de novo sterol biosynthesis and special pharmacokinetic properties, can induce radical parasitological cure of both acute and chronic Chagas experimental disease. These compounds are active against nitrofuran- and nitroimidazole-resistant T.cruzi strains and maintain their activity even if the hosts are immunosuppressed and are thus logical candidates for clinical trials with Chagas disease patients. Inhibitors of cruzipain, a cathepsin L-like protease responsible for the major proteolytic activity in all stages of the life cycle of the parasite, can selectively block the proliferation of T.cruzi, both in vitro and in vivo and have curative activity in murine models of acute Chagas disease; a significant effort is being devoted to their development as antiparasitic drugs. Alkyl-lysophospholipids, which selectively block phosphatidyl-choline biosynthesis in T.cruzi, are promising antiparasitic agents with good oral activity and low toxicity. Other biochemical pathways have been identified as potential chemotherapeutic targets, including hypoxanthine guanine phosphoribosyl transferase and the enzymes involved in the synthesis and metabolism of trypanothione and inorganic pyrophosphate. PMID- 11860368 TI - Cell biology of Leishmania spp.: invading and evading. AB - Parasitic protozoa of the genus Leishmania infect mammalian mononuclear phagocytic cells causing a potentially fatal disease with a broad spectrum of clinical manifestations. The drugs of choice used in the leishmaniasis therapy are significantly toxic, expensive and faced with a growing frequency of refractory infections. Thus the search for new leishmanicidal compounds is urgently required. In order to perform a proper drug design and to understand the modes of action of such compounds it is necessary to elucidate the intricate cellular and molecular events that orchestrate the parasite biology. In order to invade the host cell Leishmania are able to recruit different surface receptors which may assist engaging the microbicidal responses. In the intracellular milieu these pathogens can deactivate and/or subvert the phagocyte signal transduction machinery rendering these cells permissive to infection. In the present review we attempted to approach some of the most relevant cellular and biochemical invasion and evasion strategies employed by Leishmania parasites. PMID- 11860369 TI - Chemotherapy of leishmaniasis. AB - Leishmaniasis, in its variety of visceral (VL), cutaneous (CL) and mucocutaneous (MCL) forms, directly affects about 2 million people per annum, with approximately 350 million individuals at risk worldwide. During the last 10 years there have been extensive epidemics of the visceral form of the disease, which is also emerging as an important opportunistic infection in immunocompromised patients, especially those co-infected with HIV. The control of leishmaniasis remains a problem principally a zoonotic infection, except in epidemics where it is anthroponotic, interruption of transmission is difficult, though not impossible. No vaccines exist for either VL, CL or MCL and chemotherapy is inadequate and expensive. Current regimes use pentavalent antimony as primary therapy, which must be administered parenterally. Should this fail, a number of other drugs may be employed, depending upon the species of Leishmania concerned and the resources available to the health professionals involved. Recommended secondary treatment employs a variety of drugs, again depending on the nature of the infection. The most widely used of these is amphotericin B, which is highly active but has extensive toxicity complications. The newer formulations of this drug are too expensive to use for the majority of endemic countries. Pentamidine and paromomycin are used in some instances, and a new anti-leishmanial, miltefosine, may be used in the future. In short, there remains a pressing need for new anti-leishmanials and this chapter reviews the current status of chemotherapy, the various avenues being investigated by researchers and their potential application in the future. PMID- 11860370 TI - Atrial distension, haemodilution, and acute control of renin release during water immersion in humans. AB - We tested the hypothesis that atrial distension (stimulation of cardiopulmonary baroreceptors) is not the single pivotal stimulus for the acute suppression of renin release during water immersion in humans and that immersion-induced haemodilution constitutes an important additional stimulus. In nine healthy male subjects, identical increases in atrial distension were induced by two immersion procedures (of 30 min each); one without (WI) and one with attenuation (WI + cuff) of the concomitant haemodilution (estimated from changes in plasma protein concentration) by inflating thigh cuffs during immersion. During WI, central venous pressure (CVP) and left atrial diameter (LAD) increased (P < 0.05) by 5.5 +/- 0.4 mmHg and 4.6 +/- 0.5 mm, respectively, and plasma protein concentration and plasma renin activity (PRA) progressively decreased (P < 0.05) by 4.8 +/- 0.5 g L(-1) and 1.6 +/- 0.2 ng mL(-1) h(-1) (to 49 +/- 4% of baseline values), respectively. The WI + cuff caused similar atrial distension as WI (CVP and LAD increased by 6.9 +/- 0.5 mmHg and 5.5 +/- 0.5 mm, respectively), attenuated haemodilution (plasma protein concentration decreased by 1.9 +/- 0.4 g L(-1), P < 0.05 vs. WI), and markedly inhibited suppression of PRA, which decreased by 0.4 +/- 0.1 ng mL(-1) h(-1) (to 87 +/- 4% of baseline values, P < 0.05 vs. WI). Differences in renin release could not be accounted for by differences in mean arterial pressure. In conclusion, baroreceptor stimulation induced by atrial distension is not the single pivotal stimulus for the acute suppression of renin release in response to intravascular volume expansion by water immersion in humans. Haemodilution constitutes a significant and conceivably the principal stimulus for the acute immersion-induced suppression of renin-angiotensin system activity. PMID- 11860371 TI - Circulatory responses to a meal in patients with a newly transplanted heart. AB - It is well established that consumption of a meal releases a gradually developing and quite marked increase in blood flow to the gastrointestinal organs and a similar and simultaneous increase in cardiac output (CO). It is not known through which mechanism the pumping of the heart adjusts so accurately to the gastrointestinal flow increase. We have approached this problem by serving a standardized, mixed meal to five patients with recently transplanted and thus denervated hearts and to five sex- and age-matched controls. Pre- and postprandial levels of CO and blood flow in the superior mesenteric artery (SMA) were recorded with Doppler ultrasound technique. The patients with transplanted hearts had significantly higher preprandial levels of heart rate (HR) and CO than the controls. With a timing similar to that seen in the controls did all five patients develop considerable and synchronous postprandial increases in superior mesenteric arterial flow and in CO. Increases in superior mesenteric arterial flow were significantly greater than the controls. Also, COs, high even before meals were given, increased further and to the same relative extent as in the control persons. The marked postprandial increase in CO, probably secondary to the increase in intestinal blood flow, could hardly come about through any sort of nervous reflex to the recently transplanted and denervated hearts. It appears more likely that a humoral connection of some sort exists between the two circulatory events. PMID- 11860372 TI - Nitric oxide-induced signalling in rat lacrimal acinar cells. AB - The aim of the present study was to investigate the physiological role of nitric oxide (NO) in mediating secretory processes in rat lacrimal acinar cells. In addition, we wanted to determine whether the acinar cells possess endogenous nitric oxide synthase (NOS) activity by measuring NO production using the fluorescent NO indicator 4,5-diaminofluorescein (DAF-2). We initiated investigations by adding NO from an external source by means of the NO-donor, S nitroso-N-acetyl-penicillamine (SNAP). Cellular concentrations of cyclic guanosine 5'-phosphate (cGMP) ([cGMP]) were measured by radioimmunoassay (RIA), and we found that SNAP induced a fast increase in the [cGMP], amounting to 350% of the [cGMP] in resting cells. Moreover, addition of SNAP and elevating [cGMP] in fura-2 loaded lacrimal acinar cells, resulted in a cGMP-dependent protein kinase-mediated release of Ca2+ from intracellular stores, leading to a rise in the intracellular free Ca2+ concentration ([Ca2+]i). The Mn2+ quenching studies revealed that the Ca2+ release was not accompanied by Ca2+ influx. Finally, we demonstrate that lacrimal acinar cells possess endogenous NOS activity, which is activated by beta-adrenergic stimulation and not by a rise in [Ca2+]i alone. We show that in rat lacrimal acinar cells, NO and cGMP induce Ca2+ release from intracellular stores via G kinase activation. However, the changes in [Ca2+]i are relatively small, suggesting that this pathway plays a modulatory role in Ca2+ signalling, thus not by itself causing fast transient increases in [Ca2+]i. In addition, we suggest that endogenously produced NO activated by beta-adrenergic receptor stimulation, plays an important role in signalling to the surrounding tissue. PMID- 11860373 TI - Acetazolamide inhibits stimulated feline liver and gallbladder bicarbonate secretion. AB - Bile acidification is a key factor in preventing calcium carbonate precipitation and gallstone formation. Carbonic anhydrase II (CA II), that is inhibited by acetazolamide, plays a role in regulation of the acid-base balance in many tissues. This study examines the effect of acetazolamide on secretin- and vasoactive intestinal peptide (VIP)-stimulated gallbladder mucosal bicarbonate and acid secretion. Gallbladders in anaesthetized cats were perfused with a bicarbonate buffer bubbled with CO2 in air. In 20 experiments VIP (10 microg kg( 1) h(-1)) and in 10 experiments secretin (4 microg kg(-1) h(-1)) were infused continuously intravenous (i.v.). Hepatic bile and samples from the buffer before and after perfusion of the gallbladder were collected for calculation of ion and fluid transport. During basal conditions a continuous secretion of H+ by the gallbladder mucosa was seen. Intravenous infusion of vasoactive intestinal peptide (VIP) and secretin caused a secretion of bicarbonate from the gallbladder mucosa (P < 0.01). This secretion was reduced by intraluminal (i.l.) acetazolamide (P < 0.01). Bile flow was enhanced by infusion of VIP and secretin (P < 0.01) but this stimulated outflow was not affected by i.v. acetazolamide. The presence of CA II in the gallbladder was demonstrated by immunoblotting. Biliary CA activity has an important function in the regulation of VIP- and secretin-stimulated bicarbonate secretion across the gallbladder mucosa. PMID- 11860374 TI - Glycine-extended gastrin-17 stimulates acid secretion only via CCK-2 receptor induced histamine release in the totally isolated vascularly perfused rat stomach. AB - The effects of gastrin precursors have been discussed during recent years. However, the mechanism for their action, whether through a novel receptor on the parietal cell or a cholecystokinin-2 (CCK-2) receptor on the enterochromaffin like (ECL) cells, is still not settled. This study examines the effect of glycine extended gastrin-17 (Gly-G-17), the main non-amidated gastrin precursor, on gastric acid secretion and histamine release in the totally isolated vascularly perfused rat stomach. Glycine-extended gastrin-17 at the concentrations from 0.52 to 520 nmol L(-1) was administered to the totally isolated vascularly perfused rat stomach. Glycine-extended gastrin-17 at 52 or 520 nmol L(-1), and gastrin-17 at 0.52 nmol L(-1)were co-administered to examine whether glycine-extended gastrin augmented maximal gastrin stimulated acid secretion and histamine release. Both Gly-G-17 at 52 nmol L(-1) and gastrin-17 (G-17) at 0.52 nmol L(-1) were administered together with the histamine-2 receptor antagonist ranitidine at 10 micromol L(-1). Gastric acid and venous histamine output were measured. Glycine-extended gastrin-17 at lower concentrations from 0.52 to 5.2 nmol L(-1) did not stimulate gastric acid output or histamine release, whereas higher concentrations from 52 to 520 nmol L(-1) elicited a concentration-dependent increase in acid secretion and histamine release. The outputs of acid and histamine at 520 nmol L(-1) Gly-G-17 were at the same level as those found for G 17 at its maximally effective concentration of 0.52 nmol L(-1). Glycine-extended gastrin-17 at maximally effective concentration of 520 nmol L(-1) did not augment maximal gastrin stimulated acid secretion or histamine release. Ranitidine inhibited G-17 and Gly-G-17 stimulated acid secretion to a similar degree. This study confirms that the stimulatory effect of Gly-G-17 on gastric acid secretion is via a CCK-2 receptor on the ECL cell. PMID- 11860375 TI - Differential effect of oestrogen on post-exercise cardiac muscle myeloperoxidase and calpain activities in female rats. AB - The effects of oestrogen administration on 1 h post-exercise cardiac muscle myeloperoxidase (MPO) and calpain activities were determined in female rats. Rats were ovariectomized and implanted for 2 weeks with either oestrogen (25 mg 17 oestradiol) or placebo pellets or left with ovaries intact. Rats were then run for 1 h at 21 m min-1, 12% grade, killed 1 h post-exercise and cardiac muscle and blood samples were removed. Control animals from each group were killed without prior exercise. Serum oestrogen levels in the order of the highest to lowest were; ovariectomized oestrogen replaced rats > intact ovaries rats > ovariectomized placebo rats. Oestrogen induced significant (P < 0.05) elevations in cardiac MPO activity at rest and at 1 h post-exercise in ovariectomized rats. No significant elevations in cardiac MPO activity were evident in placebo ovariectomized or normal ovary rats at rest or post-exercise. Cardiac calpain activities were similar in all unexercised groups. Ovariectomized placebo and intact ovary rats had significantly (P < 0.05) elevated cardiac calpain activities 1 h post-exercise while calpain activity was not significantly elevated in hearts from ovariectomized oestrogen rats. These results demonstrate that oestrogen supplementation in ovariectomized rats induces elevations in cardiac muscle MPO activities at rest and at 1 h post-exercise. This is opposite to the effect of oestrogen in post-exercise skeletal muscle and implies a greater neutrophil infiltration into cardiac muscle caused by oestrogen. This effect cannot be explained by changes in 1 h post-exercise cardiac muscle calpain activity, the elevation of which was suppressed by oestrogen administration. Oestrogen influences cardiac calpain activity similarly to its effect in skeletal muscle. Thus, oestrogen administration to ovariectomized rats induces elevations in cardiac MPO activity while suppressing cardiac calpain activity. PMID- 11860376 TI - Responses of human elbow flexor muscles to electrically stimulated forced lengthening exercise. AB - This study developed an electrical stimulation model for human elbow flexors to examine eccentric exercise-induced muscle damage and adaptation. Male students (n=17) were randomly placed into one of two groups; isometric (ES-ISO, n=8) and eccentric (ES-ECC, n=9). The elbow joint was fixed at 90 degrees (1.57 rad) and the elbow flexors stimulated percutaneously by an electronic muscle stimulator for 5 s through two electrodes placed over the muscles for ES-ISO. In ES-ECC, the muscles were stimulated similarly to the ES-ISO, but the elbow joint was forcibly extended from an elbow flexed (90 degrees 1.57 rad) to a full-extended position (180 degrees, 3.14 rad) in 3 s. Maximal voluntary isometric force, range of motion, upper arm circumference, muscle thickness by ultrasonography, muscle soreness, plasma creatine kinase and aspartate aminotransferase activities were assessed before and for 4 days after exercise. ES-ECC produced significantly larger changes in all criterion measures compared with ES-ISO (P < 0.01). These findings confirmed that eccentric muscle actions induced muscle damage, but isometric contractions resulted in little or no damage. Six subjects from the ES ECC group repeated the same eccentric exercise (ECC2) 2 weeks after the first bout (ECC1), and changes in the criterion measures were compared between the bouts. Changes in all criterion measures after ECC2 were significantly smaller than ECC1 (P < 0.01). These results suggest that the first eccentric exercise produced a protective effect against muscle damage in the subsequent eccentric exercise bout, which does not involve adaptations in the central nervous system. PMID- 11860379 TI - Responsive interventions need responsible funding. PMID- 11860378 TI - Toward more responsive and effective intervention systems for alcohol-related problems. PMID- 11860380 TI - Treatment services for drinking problems: hidden questions. PMID- 11860381 TI - Ask the right questions and make good use of the answers: a response to Humphreys & Tucker. PMID- 11860383 TI - Scribbling in the margins: comments on Humphreys & Tucker. PMID- 11860382 TI - Striking the balance between science and common sense. PMID- 11860385 TI - Conversation with Enoch Gordis. PMID- 11860386 TI - Drug stimulus generalization and Gossop's 'Web of dependence'. PMID- 11860387 TI - Applying extinction research and theory to cue-exposure addiction treatments. AB - AIMS: To evaluate the efficacy of cue-exposure addiction treatment and review modern animal learning research to generate recommendations for substantially enhancing the effectiveness of this treatment. DESIGN: Meta-analysis of cue exposure addiction treatment outcome studies (N=9), review of animal extinction research and theory, and evaluation of whether major principles from this literature are addressed adequately in cue-exposure treatments. FINDINGS: The meta-analytical review showed that there is no consistent evidence for the efficacy of cue-exposure treatment as currently implemented. Moreover, procedures derived from the animal learning literature that should maximize the potential of extinction training are rarely used in cue-exposure treatments. CONCLUSIONS: Given what is known from animal extinction theory and research about extinguishing learned behavior, it is not surprising that cue-exposure treatments so often fail. This paper reviews current animal research regarding the most salient threats to the development and maintenance of extinction, and suggests several major procedures for increasing the efficacy of cue-exposure addiction treatment. PMID- 11860388 TI - Dual dependence: assessment of dependence upon alcohol and illicit drugs, and the relationship of alcohol dependence among drug misusers to patterns of drinking, illicit drug use and health problems. AB - AIMS: The study investigates severity of alcohol dependence among drug misusers. Specifically, it investigates the inter-relationship of alcohol and drug dependence and associations with alcohol consumption, drug consumption and substance-related problems. DESIGN, SETTING, PARTICIPANTS: The sample comprised 735 people seeking treatment for drug misuse problems, who were current (last 90 days) drinkers. MEASUREMENTS: Data were collected by structured face-to-face interviews. Dependence upon illicit drugs and upon alcohol was measured by the Severity of Dependence Scale (SDS). FINDINGS: Three groups of drinkers were identified: non-alcohol-dependent drug misusers (63%); low-dependence (19%); and high-dependence (18%). Many drug misusers were drinking excessively and alcohol dependence was related to patterns of alcohol and drug consumption. High dependence drinkers were more likely to drink extra-strength beer; they were less frequent users of heroin and crack cocaine but more frequent users of benzodiazepines, amphetamines and cocaine powder; they reported more psychological and physical health problems. The SDS was found to have good reliability and validity as a measure of alcohol dependence. SDS scores for alcohol and drug dependence were unrelated. CONCLUSIONS: Alcohol use is an important and under-rated problem in the treatment of drug misusers. A comprehensive assessment of alcohol use among drug misusers should include separate assessments of alcohol consumption, alcohol-related problems and severity of alcohol dependence. PMID- 11860389 TI - Heroin use in New South Wales, Australia, 1996-2000: 5 year monitoring of trends in price, purity, availability and use from the Illicit Drug Reporting System (IDRS). AB - AIMS: To document trends in the price, purity, availability and use of heroin in New South Wales detected by the Illicit Drug Reporting System (IDRS) between 1996 and 2000, and to demonstrate the utility of the IDRS in identifying such trends. DESIGN: The IDRS compares information derived from interviews with injecting drug users, key informants who work in the illicit drugs field, and key indicator data on illicit drug trends. SETTING: New South Wales, Australia. FINDINGS: The price of heroin approximately halved over this period, from a median of A$400 per gram in 1996 to A$220 per gram in 2000. While the price of heroin fell dramatically over the study period, the purity of police seizures of the drug was high across all years, ranging between 62% and 71%. In all years heroin was considered easy to obtain by both heroin users who purchased the drug, and by key informants from the law enforcement and health fields. Concurrent with the large fall in heroin prices, there appeared to have been an increase in the number of heroin users. Between 1997 and 1998 there was a sharp increase in the injecting use of cocaine by heroin users in NSW, a pattern that has persisted. CONCLUSIONS: Regular and formal monitoring of illicit drug trends provides timely data in a systematic way to inform health and law enforcement policies towards current and emerging illicit drug problems. PMID- 11860390 TI - Cannabis dependence in young adults: an Australian population study. AB - OBJECTIVES: The symptomatology of cannabis dependence remains ill-defined and its importance controversial. Compared with alcohol dependence, the symptom profile of cannabis dependence has received little attention. We aimed to (a) report cannabis use in a representative population of young adults, (b) examine cannabis dependence symptoms according to frequency in the dependence syndrome and (c) contrast the symptomatology of cannabis and alcohol syndromes. METHODS: 1601 young adults (mean age 20.7 years) from an Australian longitudinal cohort study (N=2032) were surveyed in 1998. Regular substance users were assessed for DSM-IV cannabis and alcohol dependence. Prevalence estimates allowed for sampling variation and attrition. RESULTS: Fifty-nine per cent reported life-time use of cannabis, 17% used at least weekly and 7% (11% males, 4% females) met criteria for cannabis dependence. Symptom prevalence in dependent cannabis users was: persistent desire 91%; unintentional use 84%; withdrawal 74%; excessive time obtaining/using 74%; continued use despite health problems 63%; tolerance 21%; and social consequences 18%. The combination of withdrawal, persistent desire and unintentional use was reported by 57%. Dependent cannabis users reported compulsive and out-of-control use more frequently than dependent alcohol users, withdrawal similarly and tolerance considerably less often. CONCLUSIONS: Cannabis use appears to be normative behaviour in young Australians. Progression beyond weekly use of cannabis carries a significant risk of dependence that should be considered in the public health response. The differing profiles of cannabis and alcohol dependence, particularly with regard to craving, draws attention to the need for further study of cannabis dependence as an important and distinct disorder in young adults. PMID- 11860391 TI - Use of anabolic steroids and associated health risks among gay men attending London gyms. AB - AIM: To examine, by HIV status, the use of anabolic steroids among London gay men and their effect on physical and mental health. DESIGN: Cross-sectional survey using self-administered questionnaire. SETTING: Six gyms in central London. PARTICIPANTS: 772 gay men using the gyms in January-February 2000. MEASUREMENTS: Proportion of gay men who reported (i) using steroids, (ii) sharing injecting equipment, (iii) experiencing side-effects and (iv) having suicidal thoughts or feeling depressed. FINDINGS: Of 772 gay men, 117 (15.2%) had used and 90 (11.7%) had injected steroids in the previous 12 months: HIV positive men (steroid use) 31.7% (40/126), HIV negative men 14.5% (69/477), never-tested for HIV 4.7% (8/169) (p < 0.001). No one reported sharing needles or syringes. HIV positive men were more likely to have used steroids for medical reasons than other men (24.3% versus 5.9%, p=0.01). Nearly all steroid users (96.4%) reported side effects including testicular atrophy (51.0%), insomnia (47.7%), depression between cycles (25.2%) and hypertension (19.0%). Steroid users were more likely to have had suicidal thoughts in the previous 6 months than non-users (22.6% versus 11.2%, adjusted odds ratio after controlling for HIV status 1.84; 95% CI 1.10-3.12, p=0.02) or to have felt depressed (49.1% versus 38.5%, adjusted OR 1.52, 95% CI 1.01-2.30, p=0.047) CONCLUSION: One in seven gay men surveyed in central London gyms in 2000 said they had used steroids in the previous 12 months. HIV positive men were more likely to have used steroids than other men, some therapeutically. Side effects were reported widely and steroid use was associated with having had suicidal thoughts and feeling depressed, although cause and effect could not be established. Our findings suggest that steroid use among gay men may have serious consequences for both physical and mental health. PMID- 11860392 TI - Smoking cessation among daily smokers, aged 45-69 years: a longitudinal study in Malmo, Sweden. AB - OBJECTIVE: To investigate differences in snuff consumption, socio-demographic and psychosocial characteristics between baseline daily smokers who had remained daily smokers, become intermittent smokers or stopped smoking at the 1-year follow-up. DESIGN, SETTING, PARTICIPANTS AND MEASUREMENTS: A population of 12 507 individuals aged 45-69 years, interviewed at baseline in 1992-94 and at a 1-year follow-up, was investigated in this longitudinal study. The three groups of baseline daily smokers were compared to the total population according to socio demographic, psychosocial and snuff consumption characteristics. A multivariate logistic regression model was used to assess differences in psychosocial conditions, adjusting for age, sex, country of origin, marital status, education and snuff consumption. FINDINGS: Eighty-six per cent of all baseline daily smokers remained daily smokers, 6.5% had become intermittent smokers and 7.3% had stopped smoking at the 1-year follow-up. The daily smokers who remained daily smokers were more likely to be born in other countries than Sweden, not married, have a lower educational level and poorer psychosocial conditions than the total population, while the socio-demographic characteristics and psychosocial resources of those daily smokers who had become intermittent smokers or had stopped smoking were much more similar to the general population, with the exception of a higher snuff consumption, especially for intermittent smokers. CONCLUSIONS: Daily smokers who remained daily smokers at the 1-year follow-up had poorer psychosocial assets, especially social participation, than baseline daily smokers who had become intermittent smokers or had stopped smoking, and the general population. The results suggest that low levels of social participation are a potent barrier against smoking cessation. Snuff consumption may explain a part of the increase in smoking cessation among men as opposed to women in Sweden. PMID- 11860393 TI - Effects of paternal drinking, conduct disorder and childhood home environment on the development of alcohol use disorders in a Thai population. AB - AIMS: To identify influences on the development of alcohol use disorders in a Thai population, particularly parental drinking and childhood environment. DESIGN: Case-control study. SETTING: A university hospital, a regional hospital and a community hospital in southern Thailand. PARTICIPANTS: Ninety-one alcohol dependents and 77 hazardous/harmful drinkers were recruited as cases and 144 non- or infrequent drinkers as controls. MEASUREMENTS: Data on parental drinking, family demographic characteristics, family activities, parental disciplinary practice, early religious life and conduct disorder were obtained using a structured interview questionnaire. The main outcome measure was the subject's classification as alcohol-dependent, hazardous/harmful drinker or non-/infrequent drinker. FINDINGS: A significant relationship was found between having a drinking father and the occurrence of hazardous/harmful drinking or alcohol dependence in the subjects. Childhood factors (conduct disorder and having been a temple boy, relative probability ratios, RPRs and 95% CI: 6.39, 2.81-14.55 and 2.21, 1.19 4.08, respectively) also significantly predicted alcohol dependence, while perceived poverty and ethnic alienation was reported less frequently by hazardous/harmful drinkers and alcohol-dependents (RPRs and 95% CIs=0.34, 0.19 0.62 and 0.59, 0.38-0.93, respectively) than the controls. The relative probability ratio for the effect of the father's infrequent drinking on the son's alcohol dependence was 2.92 (95% CI=1.42-6.02) and for the father's heavy or dependent drinking 2.84 (95% CI=1.31-6.15). CONCLUSIONS: Being exposed to a light drinking father increases the risk of a son's alcohol use disorders exhibited either as hazardous-harmful or dependent drinking. However, exposure to a heavy- or dependent-drinking father is associated more uniquely with an increased risk of his son being alcohol-dependent. The extent to which this is seen in other cultures is worthy of exploration. PMID- 11860394 TI - A fair hearing or academic kangaroo court? PMID- 11860395 TI - Raising the profile of human research ethics in addictions research: a key role for addictions journals. PMID- 11860396 TI - Alcohol consumption and stroke: difficulties in assessing the relationships. PMID- 11860397 TI - Ecstasy tablet testing: a case of guilty until proven innocent? PMID- 11860400 TI - Review article: Immunosuppressants in distal ulcerative colitis. AB - BACKGROUND: Distal ulcerative colitis may prove to be resistant to steroids and aminosalicylates, but total colectomy is more difficult to justify than in severe extensive colitis. Immunosuppression is of established benefit in generalized colitis, but there are no data available specific to distal disease. AIM: To determine whether the protocol-driven use of immunosuppressants in resistant distal ulcerative colitis is of similar efficacy and safety to that in extensive disease. METHODS: Two hundred and twenty-eight patients with distal ulcerative colitis seen in a 5-year period were identified from a prospective database. Details of 52 who had received immunosuppression were analysed. RESULTS: The 52 patients received 68 courses of therapy (53 azathioprine, five mercaptopurine, 10 ciclosporin). The thiopurines yielded clinically valuable responses in only 43% of courses, with failure of response in 16% and toxicity in 34%. Ciclosporin was helpful on only two of 10 occasions. Eight patients required total colectomy. Adverse events were typical of those normally associated with immunosuppressants, with potential risk to life in seven patients; treatment was discontinued because of toxicity on a total of 31% of occasions. CONCLUSIONS: Immunosuppression appears to be of lower efficacy and higher toxicity in resistant distal colitis than when used in more extensive colitis. PMID- 11860401 TI - Review article: Single subject trials as a research instrument in gastrointestinal pharmacology. AB - The single subject trial is a randomized controlled trial carried out in the individual patient, and the result obtained is specific to the individual patient and the drug being investigated. This type of trial offers a supplement to traditional parallel group trials, especially in patients with heterogeneous disorders, often characterized by varying treatment responses and/or high placebo response rates. Pooled results from several individual single subject trials could extend the conclusions beyond the individual patient, and help to characterize a subset of responders to a specific treatment or clarify the heterogeneity of the disease. The basic principles of the different single subject trial designs are described. Advantages and limitations are reviewed with a special focus on published trials in functional gastrointestinal disorders. The single subject trial may be a valuable supplement to traditional drug treatment trials, either used as isolated trials in individual patients to determine optimal therapy, or in groups of patients to identify those with a uniform response to treatment. However, the lack of validation and reliability studies limits the value of the single subject trials presented so far. PMID- 11860399 TI - Current concepts in the management of Helicobacter pylori infection--the Maastricht 2-2000 Consensus Report. AB - Significant progress and new insights have been gained in the 4 years since the first Maastricht Consensus Report, necessitating an update of the original guidelines. To achieve this, the European Helicobacter Pylori Study Group organized a meeting of specialists and experts from around the world, representatives from National Gastroenterology Societies and general practitioners from Europe to establish updated guidelines on the current management of Helicobacter pylori infection. The meeting took place on 21-22 September 2000. A "test and treat" approach is recommended in adult patients under the age of 45 years (the age cut-off may vary locally) presenting in primary care with persistent dyspepsia, having excluded those with predominantly gastro-oesophageal reflux disease symptoms, non-steroidal anti-inflammatory drug users and those with alarm symptoms. Diagnosis of infection should be by urea breath test or stool antigen test. As in the previous guidelines, the eradication of H. pylori is strongly recommended in all patients with peptic ulcer, including those with complications, in those with low-grade gastric mucosa-associated lymphoid tissue lymphoma, in those with atrophic gastritis and following gastric cancer resection. It is also strongly recommended in patients who are first degree relatives of gastric cancer patients and according to patients' wishes after full consultation. It is advised that H. pylori eradication is considered to be an appropriate option in infected patients with functional dyspepsia, as it leads to long-term symptom improvement in a subset of patients. There was consensus that the eradication of H. pylori is not associated with the development of gastro-oesophageal reflux disease in most cases, and does not exacerbate existing gastro-oesophageal reflux disease. It was agreed that the eradication of H. pylori prior to the use of non-steroidal anti-inflammatory drugs reduces the incidence of peptic ulcer, but does not enhance the healing of gastric or duodenal ulcer in patients receiving antisecretory therapy who continue to take non-steroidal anti-inflammatory drugs. Treatment should be thought of as a package which considers first- and second-line eradication therapies together. First-line therapy should be with triple therapy using a proton pump inhibitor or ranitidine bismuth citrate, combined with clarithromycin and amoxicillin or metronidazole. Second-line therapy should use quadruple therapy with a proton pump inhibitor, bismuth, metronidazole and tetracycline. Where bismuth is not available, second-line therapy should be with proton pump inhibitor-based triple therapy. If second-line quadruple therapy fails in primary care, patients should be referred to a specialist. Subsequent failures should be handled on a case-by-case basis by the specialist. In patients with uncomplicated duodenal ulcer, eradication therapy does not need to be followed by further antisecretory treatment. Successful eradication should always be confirmed by urea breath test or an endoscopy-based test if endoscopy is clinically indicated. Stool antigen test is the alternative if urea breath test is not available. PMID- 11860402 TI - Antioxidant effects of herbal therapies used by patients with inflammatory bowel disease: an in vitro study. AB - BACKGROUND: Herbal remedies used by patients for treatment of inflammatory bowel disease include slippery elm, fenugreek, devil's claw, Mexican yam, tormentil and wei tong ning, a traditional Chinese medicine. Reactive oxygen metabolites produced by inflamed colonic mucosa may be pathogenic. Aminosalicylates (5-ASA) are antioxidant and other such agents could be therapeutic. AIMS: To assess the antioxidant effects of herbal remedies in cell-free oxidant-generating systems and inflamed human colorectal biopsies. METHODS: Luminol-enhanced chemiluminescence in a xanthine/xanthine oxidase cell-free system was used to detect superoxide scavenging by herbs and 5-ASA, and fluorimetry to define peroxyl radical scavenging using a phycoerythrin degradation assay. Chemiluminescence was used to detect herbal effects on generation of oxygen radicals by mucosal biopsies from patients with active ulcerative colitis. RESULTS: Like 5-ASA, all herbs, except fenugreek, scavenged superoxide dose dependently. All materials tested scavenged peroxyl dose-dependently. Oxygen radical release from biopsies was reduced after incubation in all herbs except Mexican yam, and by 5-ASA. CONCLUSIONS: All six herbal remedies have antioxidant effects. Fenugreek is not a superoxide scavenger, while Mexican yam did not inhibit radical generation by inflamed biopsies. Slippery elm, fenugreek, devil's claw, tormentil and wei tong ning merit formal evaluation as novel therapies in inflammatory bowel disease. PMID- 11860403 TI - Evaluation of renal function following treatment with 5-aminosalicylic acid derivatives in patients with ulcerative colitis. AB - BACKGROUND: A number of cases of nephrotoxicity have been reported in patients with inflammatory bowel disease taking oral 5-aminosalicylic acid (5-ASA). AIM: To evaluate the effects of 9 months of therapy with mesalazine or olsalazine on renal function in patients with ulcerative colitis in remission. METHODS: Forty patients with ulcerative colitis in complete remission for 6 months were randomized to either olsalazine (n=20) or mesalazine (n=20 for nine months). Thirty-six of the 40 patients were on prior salicylate therapy. Disease activity was the measure ofclinical efficacy and was assessed by the Harvey-Bradshaw Index (HBI). Laboratory efficacy measurements included glomerular filtration rate (GFR), microalbuminuria, urinary gluthathione S-transferase (GST) and serum C reactive protein (CRP). Safety analysis consisted of documentation of adverse events and laboratory values. RESULTS: There was no significant reduction in the GFR overall on therapy. The levels of GFR adjusted for baseline were similar in the two treatment groups after 3, 6 and 9 months. A significantly higher percentage of mesalazine-treated patients experienced drug related adverse events, all of a minor nature. The incidence of adverse events causing early withdrawal was similar in the two treatment groups. CONCLUSION: Treatment with mesalazine or olsalazine for 9 months had no significant impact on GFR. PMID- 11860404 TI - Effects of meal consistency and ingested fluid volume on the intragastric distribution of a drug model in humans--a magnetic resonance imaging study. AB - BACKGROUND: Controlled delivery of drugs to the small intestine in relation to emptying of an ingested meal is important in various pathophysiological conditions. We investigated the effects of different food consistencies and the amount of co-ingested liquid on the intragastric distribution of a contrast marker. METHODS: Five healthy subjects received four meals (each 650 kcal: A, mashed potato with 100 mL water; B, rice with 100 mL water; C, hamburger meal with 100 mL water; D, hamburger meal with 300 mL water). A capsule filled with gadolinium tetra-azacyclododecane tetra-acetic acid solution (as contrast marker) was ingested following meal termination, and its intragastric distribution was assessed by magnetic resonance imaging. RESULTS: Initially, marker distribution was confined to the fundus, and subsequently extended along the inner curvature of the stomach. The maximum distribution volume of the marker was lower in meal A than in meal B (P < 0.05). No differences in marker distribution were observed when the hamburger meal was given with 100 or 300 mL water. CONCLUSIONS: The intragastric distribution kinetics of the marker gadolinium tetra azacyclododecane tetra-acetic acid appeared to depend on meal consistency, but not on the amount of water co-ingested. Three-dimensional magnetic resonance imaging allows detailed analysis of the intragastric distribution of a drug model in relation to meal emptying and intragastric meal distribution. PMID- 11860405 TI - Effects of 5-HT(3) antagonism on postprandial gastric volume and symptoms in humans. AB - BACKGROUND: Alosetron reduces symptoms of dyspepsia, but the physiological basis for the symptomatic benefit is unclear. AIM: To assess 5-HT3 antagonism on postprandial gastric volume and symptoms after ingestion of maximum tolerable volume of a liquid meal. METHODS: In 36 healthy volunteers, we assessed effects of placebo, 0.5 and 1 mg b.d. alosetron on fasting and postprandial gastric volumes (using single photon emission computed tomography) and symptoms based on 100 mm VAS, 30 min after maximum volume ingested. RESULTS: The 5-HT3 antagonist reduced postprandial symptoms (aggregate score: P < 0.05), nausea (P < 0.001), and tended to reduce bloating (P=0.08). Both 0.5 and 1 mg alosetron reduced nausea (P < 0.025); 1 mg alosetron reduced aggregate symptoms (P < 0.05) and bloating (P < 0.05). Effects on pain (P=0.19) and fullness (P=0.14) were not statistically significant. There were no significant effects of the 5-HT3 antagonist on volume of meal tolerated or on SPECT-measured fasting or postprandial gastric volumes. CONCLUSION: 5-HT3 antagonism reduces aggregate symptoms, nausea and bloating after a liquid meal without increase in gastric volumes, suggesting a role for 5-HT3 in afferent functions in healthy humans during the postprandial period. PMID- 11860406 TI - Pharmacokinetics and biochemical effects of hepapoietin in patients with chronic liver disease. AB - BACKGROUND: Hepapoietin is a naturally occurring cytokine that promotes hepatocyte growth. Animal studies have suggested that hepapoietin and hepatocyte growth factor have a potential role in the prevention and management of liver diseases. However, human studies have been lacking. AIM: To evaluate the safety and pharmacokinetics of single escalating doses of hepapoietin in patients with chronic liver disease. METHODS: An open-label, single escalating dose trial with five different doses of hepapoietin (1, 3, 10, 30 and 100 mg) was performed. Adults with chronic, compensated, non-viral liver disease were included. Liver function tests were obtained before dosing, 24 h after hepapoietin administration and on days 4, 7, 30 and 45. All patients were followed for 45 days. RESULTS: Twenty-five subjects received hepapoietin, with five subjects each at 1, 3, 10, 30 and 100 mg of hepapoietin. Significant decreases occurred in total bilirubin, ammonia, partial thromboplastin time and cholesterol levels overall, and both high-density and low-density lipoprotein cholesterol showed a downward trend. An increase in albumin was observed at the 30 mg dose level. Slight decreases in haemoglobin and red blood cell levels were observed at day 4, but returned to normal levels immediately thereafter. Child-Pugh scores from day 0 to day 7 were improved in 24%, stable in 64% and worse in 12% of patients. Hepatic encephalopathy displayed changes from day 0 to day 45 with improvement in 16%, no change in 80% and worsening in 4%. CONCLUSIONS: Hepapoietin in doses up to 100 mg is safe for use in humans. Potential benefits are suggested by significant decreases in bilirubin, ammonia, partial thromboplastin time and cholesterol levels and an increase in albumin. Further studies with multiple dosing regimens are needed to identify the clinical utility of hepapoietin in the management of chronic liver disease. PMID- 11860407 TI - Interferon-alpha plus ribavirin for 12 months increases the sustained response rates in chronic hepatitis C relapsers. AB - BACKGROUND: The effectiveness and tolerability of combination therapy for 12 months have not been evaluated sufficiently in chronic hepatitis C relapsers to interferon. AIMS: To evaluate the sustained response to interferon plus ribavirin for 12 months in chronic hepatitis C relapsers. METHODS: We included 55 chronic hepatitis C relapsers in a 12-month treatment protocol with interferon (3 MU thrice weekly) plus ribavirin (1-1.2 g/day). The effectiveness was evaluated using serum aminotransferase and hepatitis C virus RNA levels, alanine aminotransferase normalization and viraemia clearance after 12 months, defining the end-of-treatment response, and 6 months after completion of therapy, defining the sustained response. Adverse effects were recorded. RESULTS: End-of-treatment response and sustained response were achieved in 47 (85%) and 37 (67%) patients, respectively; there were 10 (21%) relapsers after combination therapy. Predictive factors of sustained response included the genotype (non-1 95% vs. 1 48%; P < 0.001), lower viraemia (503 917 +/- 553 230 vs. 901 393 +/- 548 267 copies/mL; P < 0.005), higher alanine aminotransferase levels (137 +/- 75 vs. 103 +/- 41 IU/L; P < 0.05) and a lower gamma-glutamyl transpeptidase/alanine aminotransferase ratio (0.30 +/- 0.23 vs. 0.49 +/- 0.39; P < 0.05). Tolerance to therapy was good, with no withdrawals. CONCLUSIONS: Interferon plus ribavirin treatment for 12 months in chronic hepatitis C relapsers yields high sustained response rates and is well tolerated. The sustained response is related to a non-1 genotype, lower baseline viraemia, higher alanine aminotransferase level and a lower gamma glutamyl transpeptidase/alanine aminotransferase ratio. PMID- 11860408 TI - Expression of cyclooxygenase-2 in chronic hepatitis B and the effects of anti viral therapy. AB - BACKGROUND: Cyclooxygenase-2 may play a role in the development of hepatocellular carcinoma, but the relationship between cyclooxygenase-2 and chronic hepatitis B is unknown. AIM: To investigate the expression and cellular localization of cyclooxygenase-2 in chronic hepatitis B patients and the effects of anti-viral therapy. METHODS: Using immunohistochemistry, in situ hybridization, Western blot and reverse transcription polymerase chain reaction, protein and messenger RNA expression and cellular localization of cyclooxygenase-2 in 35 chronic hepatitis B patients were assessed. Fourteen histologically normal and non-viral-infected livers were used as controls. The cyclooxygenase-2 immunoreactivities of paired liver biopsies from 12 patients receiving anti-viral therapy were compared. RESULTS: Immunohistochemistry and in situ hybridization revealed that cyclooxygenase-2 expression was confined to hepatocytes. Patients with chronic hepatitis B had significantly higher cyclooxygenase-2 expression compared with controls. The cyclooxygenase-2 expression of hepatitis B e antigen-positive and negative chronic hepatitis B patients was not significantly different, although the necro-inflammatory activity of the latter group was significantly lower. Over expression of cyclooxygenase-2 in patients with chronic hepatitis B was further confirmed by Western blot and reverse transcription polymerase chain reaction. Twelve hepatitis B e antigen-positive chronic hepatitis B patients received anti viral therapy: lamivudine in seven and interferon in five. Despite hepatitis B e antigen seroconversion, disappearance of hepatitis B virus DNA in serum, normalization of liver enzymes and a significant reduction in necro-inflammatory activity in all 12 patients, no significant change in cyclooxygenase-2 expression was found. CONCLUSIONS: Chronic hepatitis B is associated with elevated cyclooxygenase-2 levels in hepatocytes, and the over-expression of this enzyme does not reflect inflammatory activity. Up-regulation of cyclooxygenase-2 persists after successful anti-viral therapy. PMID- 11860409 TI - The clinical and economic impact of competing management strategies for gastro oesophageal reflux disease. AB - BACKGROUND: Gastro-oesophageal reflux disease (GERD) is a common disorder in the primary care setting. Traditional management strategies consist of sequentially intensive therapeutic trials followed by invasive diagnostic testing for nonresponders. A high dose proton pump inhibitor trial (the "proton pump inhibitor test") has been shown to be an accurate diagnostic alternative, and may be an efficient initial approach to patients with GERD symptoms. AIM: To examine the clinical, economic and policy implications of alternative management strategies for GERD. METHODS: Decision analysis was used to calculate the clinical and economic outcomes of competing management strategies. The traditional strategy incorporates sequential therapeutic trials with more intensive therapy ("step-up" approach) followed by sequential invasive diagnostic testing of nonresponders. The "proton pump inhibitor test" strategy includes an initial "proton pump inhibitor test" (7 days of omeprazole; 40 mg AM + 20 mg PM daily) followed by less intensive therapeutic trials in those testing positive ("step-down" approach) with sequential invasive diagnostic testing as needed. Cost estimates were based on Medicare reimbursement and average wholesale drug prices. Probability estimates were derived from a systematic review of the published medical literature. Model results are reported as the average and incremental cost-per-symptom free patient and cost-per-quality-adjusted life years (QALYs) gained. RESULTS: The average cost per patient was 1045 US dollars for the traditional step-up management strategy, compared to 1172 US dollars for the "proton pump inhibitor test" and step-down strategy. The percentage of patients who were symptom-free at 1 year was 50% for the traditional management strategy compared to 75% for the "proton pump inhibitor test" strategy. The "proton pump inhibitor test" strategy results in QALY gains of 0.01-0.05 depending on the utility estimate employed. The incremental cost-effectiveness ratio for the "proton pump inhibitor test" strategy is 510 US dollars per additional symptomatic cure over 1 year, and between 2822-10,160 US dollars per QALY gained. The traditional management strategy resulted in a greater than 5 fold increase in the utilization of upper endoscopy, which was partially offset by a 47% reduction in the use of ambulatory 24-h oesophageal pH monitoring. The reduced effectiveness of the traditional management strategy may be attributed in part to a 118% increase in the use of "high-dose" H2RAs while reducing the use of standard dose proton pump inhibitors by only 42% and "high-dose" proton pump inhibitors by 57%. CONCLUSIONS: Based on the results of this analysis, strategies utilizing the initial PPI test followed by a "step-down" approach may result in improved symptom relief and quality of life over 1 year, and more appropriate utilization of invasive diagnostic testing at a small marginal increase in total costs. These findings warrant a prospective trial comparing these competing management strategies. PMID- 11860410 TI - Effect of Helicobacter pylori eradication on duodenal ulcer scar in patients with no clinical history of duodenal ulcer. AB - BACKGROUND: Helicobacter pylori eradication has become the standard treatment for duodenal ulcer. However, there is no relevant evidence for antibacterial treatment of the white scar stage of duodenal ulcer (duodenal ulcer scar) in patients with no past history of duodenal ulcer. AIM: To investigate whether H. pylori eradication could decrease duodenal ulcer recurrence in patients with duodenal ulcer scar and no past history of duodenal ulcer. PATIENTS AND METHODS: We prospectively enrolled 66 patients with duodenal ulcer scar: 53 were H. pylori positive and 13 were H. pylori-negative. H. pylori-positive patients were randomly assigned into two groups (two-to-one allocation): 36 patients were assigned to the treatment group and 17 to the follow-up group. Thirteen H. pylori negative patients were followed up according to the study protocol. Follow-up endoscopy was performed to evaluate ulcer scar changes and H. pylori status 6 weeks after anti-H. pylori treatment and then every 6 months for up to 30 months. RESULTS: Active duodenal ulcer recurrence was identified in seven of 23 H. pylori positive/non-cured patients (30%). There was no duodenal ulcer recurrence in 43 H. pylori-negative/cured patients (0%), which was significantly different in terms of duodenal ulcer recurrence (P=0.001). CONCLUSIONS: H. pylori eradication is effective at preventing active duodenal ulcer recurrence in patients with duodenal ulcer scar and no past history of duodenal ulcer. PMID- 11860411 TI - Helicobacter pylori, parietal cell antibodies and autoimmune gastropathy in type 1 diabetes mellitus. AB - BACKGROUND: Fifteen to 20% of type 1 diabetic patients exhibit parietal cell antibodies (PCA), which are associated with autoimmune gastritis, hypochlorhydria, iron deficiency and pernicious anaemia. AIM: To examine whether Helicobacter pylori infection could explain the high prevalence of PCA and autoimmune gastropathy in diabetes. If so, H. pylori eradication could prevent autoimmune gastritis. METHODS: In 229 type 1 diabetics (M/F: 135/94; age: 41 +/- 12 years) PCA were measured. H. pylori infection was assessed by serology, urea breath test in all and by histology (updated Sydney system) in 88 subjects. Pentagastrin tests were performed in 42 patients. RESULTS: Sixty-nine patients were PCA-positive. H. pylori infection was present in 72 patients and was negatively associated with HLA-DQA1*0103-B1*0603 (OR=0.12, P=0.015) and positively with DQA1*0501-B1*0201 (OR=1.9, P=0.032). PCA-positivity was linked to HLA-DQA1*0501-B1*0301 (OR=3.9, P=0.017). A link between H. pylori and PCA was observed when PCA-positivity was defined as a titre > or = 1/20 (OR=2.0, P=0.03), but not if > or =1/40 was the cut-off point. PCA-positivity, but not H. pylori infection, was associated with iron deficiency anaemia (OR=2.7, P=0.008), pernicious anaemia (OR= 33.5, P < 0.0001), hypochlorhydria (OR=12.1, P=0.0008) and autoimmune gastritis (OR=12.5, P < 0.0001). CONCLUSIONS: The HLA-bound susceptibility of H. pylori and PCA differed. PCA-positivity but not ongoing H. pylori infection is associated with autoimmune gastritis. Low titres of PCA might reflect H. pylori infection rather than autoimmune gastropathy. PMID- 11860412 TI - Primary resistance to antibiotics and its clinical impact on the efficacy of Helicobacter pylori lansoprazole-based triple therapies. AB - AIM: To evaluate Helicobacter pylori primary resistance and its clinical impact on the efficacy of two lansoprazole-based eradication triple therapies. METHODS: H. pylori-positive patients (n=228) were randomized to receive one of the 1-week regimens: lansoprazole 30 mg, clarithromycin 500 mg and amoxicillin 1 g (LAC), or lansoprazole 30 mg, clarithromycin 500 mg and metronidazole 500 mg (LMC), each given twice daily. H. pylori status was assessed by 13C-urea breath test and culture at diagnosis and by 13C-urea breath test 6 weeks after therapy. Antibiotic susceptibility was determined by E-test (n=98). RESULTS: The eradication rates with per protocol/ intention-to-treat analyses were: LAC (n=95/114) 83%/69% and LMC (n=96/114) 85%/72%. Primary resistance was 11% for clarithromycin, 41% for metronidazole and 0% for amoxicillin. Eradication in metronidazole-susceptible/-resistant strains was 85%/82% in LAC and 83%/63% in LMC. Significantly lower cure rates were observed in clarithromycin-resistant patients treated with LAC (95% vs. 0%, P < 0.001) and LMC (86% vs. 0%, P < 0.001). CONCLUSIONS: One-week LAC and LMC are similarly effective therapies. Clarithromycin resistance significantly affected H. pylori eradication in both regimens. PMID- 11860413 TI - Nizatidine and cisapride enhance salivary secretion in humans. AB - BACKGROUND: Salivation plays an important role in the defence of the oesophageal mucosa against gastric acidic reflux and can be evoked by cholinergic stimulation. Both nizatidine and cisapride have been reported to increase acetylcholine concentrations in the cholinergic system. AIM: To investigate the effect of nizatidine and cisapride on salivary secretion, salivary epidermal growth factor and bicarbonate output. METHODS: The salivary volume and concentration of salivary epidermal growth factor and bicarbonate were measured after the administration of nizatidine (150 mg), famotidine (20 mg) and cisapride (5 mg) in 30 male healthy volunteers. RESULTS: Basal and stimulated salivary secretions were found to be increased after the administration of nizatidine and cisapride. In contrast, salivary secretion was not increased by famotidine. Although epidermal growth factor content was not augmented, nizatidine and cisapride administration also increased the bicarbonate output in mastication stimulated saliva. CONCLUSIONS: Increased salivary secretion and bicarbonate output induced by nizatidine may be useful for the treatment of patients with gastro-oesophageal reflux disease. PMID- 11860414 TI - Minor effects of Helicobacter pylori on gastric secretion and dose of lansoprazole during long-term treatment in ZE and non-ZE acid hypersecretors. AB - BACKGROUND: Helicobacter pylori infection may increase or decrease acid secretion and may augment proton pump inhibitor efficacy. Pepsin effects have not been reported. In Zollinger-Ellison syndrome (ZE) specifically, H. pylori has been reported to decrease acid. AIM: To examine H. pylori effects on secretion and dose of medication in hypersecretors (basal acid output > 15 mmol/h) undergoing long-term treatment with individually optimized lansoprazole doses. METHODS: Sixty-five patients (47 ZE and 18 non-ZE), treated for > 3 months to 10 years, were tested every 6 months with endoscopy, gastric analysis and serum gastrin. RESULTS: Forty-three per cent were H. pylori-positive. Acid, pepsin and gastrin were not different between H. pylori-positive and H. pylori-negative patients before or during long-term lansoprazole treatment. Initially, H. pylori-positive patients required less lansoprazole than H. pylori-negative patients (68 +/- 6 vs. 96 +/- 8 mg/day), but after 3 years the doses converged (83 vs. 86 mg/day). The disappearance of H. pylori in 15 patients caused no significant changes in acid, pepsin, gastrin or lansoprazole dose in the following 4 years. CONCLUSIONS: H. pylori had no significant initial or long-term physiological or potential clinical effects on acid or pepsin secretion or gastrin in these acid hypersecretors. PMID- 11860415 TI - High rate of post-therapeutic resistance after failure of macrolide nitroimidazole triple therapy to cure Helicobacter pylori infection: impact of two second-line therapies in a randomized study. AB - BACKGROUND: The optimal second-line treatment after failed Helicobacter pylori therapy has not been established. AIMS: To ascertain whether quadruple therapy or triple therapy with omeprazole, clarithromycin and amoxicillin is the superior re treatment after triple therapy containing a macrolide and a nitroimidazole, and to determine the impact of microbial in vitro resistance. METHODS: Patients after failed triple therapy were randomly allocated to one of two 1-week second-line treatments: omeprazole, 40 mg, clarithromycin, 500 mg, and amoxicillin, 1 g, all b.d.; or omeprazole, 20 mg b.d., bismuth subsalicylate, 600 mg q.d.s., metronidazole, 400 mg t.d.s., and tetracycline, 500 mg q.d.s. Post-therapeutic Helicobacter pylori status was assessed by 13C-urea breath test at least 4 weeks after treatment. RESULTS: The study was terminated after including 84 patients. H. pylori cure rates differed significantly: omeprazole-clarithromycin amoxicillin: intention-to-treat, 43%; per protocol, 50%; omeprazole-bismuth subsalicylate-metronidazole-tetracycline: intention-to-treat, 68%; per protocol, 69%. The frequencies of resistance after first-line therapy were: metronidazole, 90%; clarithromycin, 71%; both combined, 68%. For clarithromycin resistance, H. pylori cure with omeprazole-clarithromycin-amoxicillin was 30% vs. 83% for clarithromycin susceptibility. CONCLUSIONS: Omeprazole-bismuth subsalicylate metron- idazole-tetracycline was superior to omeprazole-clarithromycin amoxicillin, but both therapies yielded unsatisfactory results. The high rate of post-therapeutic dual resistance has a negative impact on omepraz- ole clarithromycin-amoxicillin, and probably also on omeprazole-bismuth subsalicylate metronidazole-tetracycline, and limits the choice for second-line treatment. PMID- 11860416 TI - Sulphide-induced energy deficiency in colonic cells is prevented by glucose but not by butyrate. AB - BACKGROUND: In ulcerative colitis, hydrogen sulphide is postulated to impair colonocyte butyrate metabolism, leading to cellular energy deficiency and dysfunction. AIMS: To determine the effects of sulphide exposure on butyrate metabolism and adenosine triphosphate levels of HT29 colonic epithelial cancer cells, and to establish whether energy deficiency can be prevented by increased butyrate concentrations or the presence of glucose. METHODS: HT29 cells were maintained in medium containing 3 mM butyrate, 5 mM glucose, or both substrates. Oxidation rates were measured by 14CO2 release from 14C-labelled substrates. Cellular adenosine triphosphate was assayed using the luciferin/luciferase chemiluminescent method. The effects of sulphide (0-5 mM) on substrate oxidation and adenosine triphosphate levels and of increasing butyrate concentration (0-30 mM) with sulphide were observed. RESULTS: HT29 cells showed similar energy substrate usage to primary colonocyte cultures. Sulphide exposure inhibited butyrate oxidation and led to a reduction in cellular adenosine triphosphate. This fall was prevented by co-incubation with glucose, but not by increasing concentrations of butyrate. CONCLUSIONS: HT29 cells utilize butyrate as an energy substrate and represent a useful in vitro model of the effects of sulphide on colonocytes. Sulphide inhibits butyrate oxidation and leads to demonstrable energy deficiency, prevented by the presence of glucose but not by increased butyrate concentrations. PMID- 11860417 TI - The importance of core surgical training for dermatologists and dermatologic surgeons. PMID- 11860418 TI - A risk scale for predicting extensive subclinical spread of nonmelanoma skin cancer. AB - BACKGROUND: The clinical appearance of nonmelanoma skin cancer may represent only a portion of microscopic tumor invasion. OBJECTIVE: To develop a scale based on high-risk characteristics for predicting the probability of extensive subclinical spread of nonmelanoma skin cancer. METHODS: Retrospective analysis of 1095 Mohs micrographic surgical cases (MMS) yielded high-risk factors for extensive tumor spread, defined as requirement of greater-than-or-equal3 MMS layers. Predictive characteristics included: any BCC on the nose, morpheaform BCC on the cheek, neck tumors and recurrent BCC in men, location on the eyelid, temple, or ear helix, and size>10 mm. Multivariate logistic regression was applied to develop a risk index. RESULTS: Tumor characteristics were assigned point values calculated from the respective odds of extension and categorized into six risk classes with probabilities of extensive subclinical spread ranging from 10% to 56%. CONCLUSION: A risk scale simplifies and enhances prediction of extensive tumors. The associated probabilities can help to guide patient preparation and appropriate therapy. PMID- 11860419 TI - A meta-analysis of the prognostic significance of sentinel lymph node status in Merkel cell carcinoma. AB - BACKGROUND: Merkel cell carcinoma is an aggressive cutaneous neoplasm with a high propensity to metastasize to lymph nodes. OBJECTIVE: The objective of this study was to determine the prognostic significance of sentinel lymph node status in patients with Merkel cell carcinoma. METHODS: A meta-analysis of case series of patients with Merkel cell carcinoma managed with sentinel lymph node biopsy was performed. RESULTS: Forty of 60 patients (67%) had a biopsy-negative sentinel lymph node; 97% of this group had no recurrence at 7.3 months median follow-up. Twenty patients (33%) had a biopsy-positive sentinel lymph node; 33% of this group experienced local, regional, or systemic recurrence at 12 months median follow-up. Risk of recurrence or metastasis was 19-fold greater in biopsy positive patients (odds ratio, 18.9; p = 0.005). None of 15 biopsy-positive patients who underwent therapeutic lymph node dissection experienced a regional recurrence; 3 of 4 who did not receive therapeutic lymphadenectomy experienced regional recurrence. CONCLUSION: Sentinel lymph node positivity is strongly predictive of a high short-term risk of recurrence or metastasis in patients with Merkel cell carcinoma. Therapeutic lymph node dissection appears effective in preventing short-term regional nodal recurrence. Aggressive adjuvant treatment should be considered for patients with positive sentinel lymph nodes. PMID- 11860420 TI - The effect of different spot sizes on the efficacy of hair removal using a long pulsed diode laser. AB - BACKGROUND: In the last years several lasers have proven their efficacy for hair removal. However, little is known about the efficacy of varying the spot size with those lasers. OBJECTIVE: To evaluate the long-term efficacy of hair removal using a diode laser with different spot sizes. METHODS: A long-pulsed diode laser (2 x 60 msec) was used. The spot size was 8 mm, 12 mm, or 14 mm. Twenty consenting volunteers were treated three times at regular intervals of 3 weeks. The ratio of the number of hairs in the treated area to an adjacent area left untreated (control) was referred to as regrowth. RESULTS: One month after laser treatment, regrowth was 23% (8 mm), 12% (12 mm), and 13% (14 mm). After 3 months regrowth was 67% (8 mm), 54% (12 mm), and 55% (14 mm). Fifteen months after treatment 4 of 16 volunteers had a regrowth rate of less than 25%. CONCLUSION: The results provide evidence for an effective and long-lasting growth delay of hairs using the long-pulsed diode laser. The use of large spot sizes improved the growth delay of hairs measured 1 month after treatment. PMID- 11860421 TI - Meshed reversed dermal graft in patients with surgical defects of sole and scalp: technique and long-term results. AB - BACKGROUND: Defects of the sole and the scalp require a reconstruction technique that combines a high survival rate in poorly vascularized areas with maximal mechanical strain. OBJECTIVE: To describe the technique of the meshed reversed dermal graft for reconstruction and to summarize the postoperative and long-term results. METHODS: Twenty-one patients have undergone reconstruction of defects following excision of acral lentiginous melanoma of the sole or squamous cell carcinoma of the sole or scalp. One patient had undergone covering of a perforating ulcer of the foot. RESULTS: In 19 of the 21 patients the meshed reversed dermal graft survived without complications. Of the patients who had undergone successful reconstruction, 14 patients were reinterviewed: 13 reported on excellent functional and satisfying cosmetic results. CONCLUSION: The meshed reversed dermal graft is an appropriate technique for reconstruction of large defects of the sole or scalp because of its high survival rate and excellent long term functional results. PMID- 11860422 TI - Laser blepharoplasty: diamond laser scalpel compared to the free beam CO2 laser. AB - BACKGROUND: Blepharoplasty has long been employed for functional and cosmetic purposes. Different modalities have been utilized to incise and remove excess eyelid tissue in an attempt to improve surgical technique. OBJECTIVE: This study compares a free beam CO2 laser to a diamond surgical scalpel combined with a CO2 laser for performance of blepharoplasty. METHODS: Ten patients underwent bilateral upper lid blepharoplasty. One side was randomly assigned to the free beam CO2 laser technique, while the other side was performed with the diamond laser scalpel. Intraoperative surgical times and hemostasis were measured. Masked observers assessed postoperative parameters at day 1, week 1, week 2, week 4, and month 4. These assessments were analyzed clinically and statistically. RESULTS: The two modalities produced similar results for the intraoperative and postoperative parameters studied. CONCLUSION: The diamond laser scalpel has the advantage of tactile feedback and the option of use without coagulation. PMID- 11860424 TI - Hair transplantation: management of donor area. AB - BACKGROUND: Hair transplantation entails the removing of a strip of permanent hair from the occipital scalp and reimplanting it piecemeal into the recipient area. Methodologies for effecting this are reasonably complex and have undergone many changes and improvements over the years. The simplest part of hair transplantation, at least in theory, is the removal of the donor strip and closure of the resultant defect. Unfortunately, however, the improvements that have taken place in hair transplantation in general have not spilled over into improved donor area outcomes. Unsightly donor scars and fibrosis are still not only with us, but are possibly even more pervasive than ever. One explanation given for deteriorated donor sites is the harvesting of relatively greater numbers of grafts. But is this the whole story? OBJECTIVE: To describe some technical aspects of donor area management consistent with harvesting the best possible donor strip, while leaving an inconspicuous scar and preserving the viability of the residual tissues for subsequent harvesting. METHODS: In 1994 a clinical research project designed to determine what technical surgical modalities are optimal for excising and reconstructing scalp tissues was commenced. This was done with particular reference to the donor area in hair transplantation. The study involved more than 1000 scalp operations using various techniques and instrumentation, and comparing and contrasting results. RESULTS: The best results were obtained when tissues were least traumatized. Tension generated at wound closure was found to be the main culprit in determining less than optimal residual donor sites. CONCLUSION: Notably improved postharvesting donor sites are most likely to result when measures are taken to ensure minimal trauma by taking definitive steps to combat tension in the tissues. Modest undermining combined with deep plane fixation facilitates channeling of tension vector forces from at-risk superficial tissues into nonundermined tissues and deep tissues, each of which is optimally equipped to withstand the adverse consequences of tension. PMID- 11860423 TI - Reactive oxygen species activate the human elastin promoter in a transgenic model of cutaneous photoaging. AB - BACKGROUND: Generation of free radicals has been shown to play a role in cutaneous alterations resulting from ultraviolet radiation. OBJECTIVE: Cells from a previously described in vitro transgenic model of cutaneous photoaging were exposed to reactive oxygen species to determine if this results in elastin promoter activation. METHODS: Reactive oxygen species were generated using a hypoxanthine and xanthine oxidase system, and elastin promoter activation was measured using cells derived from transgenic mice containing the human elastin promoter. RESULTS: Free radical generation resulted in a greater than sixfold increase in elastin promoter activity, and this increase was blocked with the addition of catalase. CONCLUSION: Elastin promoter activation may play a role in the generation of solar elastosis in photoaged skin. Utilizing hypoxanthine and xanthine oxidase with the in vitro transgenic photoaging model results in a sensitive system for evaluating agents that may prevent oxidative damage. PMID- 11860425 TI - Application of local anesthetics in dermatologic surgery. AB - BACKGROUND: Local anesthesia is an integral aspect of cutaneous surgery. Its effects provide a reversible loss of sensation in a limited area of skin, allowing dermatologists to perform diagnostic and therapeutic procedures safely, with minimal discomfort and risk to the patient. OBJECTIVE: This article discusses the application of local anesthetics in dermatologic surgery through a brief review of the mechanism of local anesthetic activity, classification of local anesthetics, and adverse effects. The discussion will focus on the different methods of administration and techniques that may make the administration less painful to patients. METHODS: An evaluation of the literature, as well as clinical experience in the administration of local anesthetics provide the basis for this review. CONCLUSION: Local anesthesia provides a safe and effective means of decreasing patient's sensitivity to pain. Because of its critical importance to the field of dermatology, a working knowledge of local anesthetics is necessary for optimal use of these agents. PMID- 11860426 TI - Decreased tear expression with an abnormal Schirmer's test following botulinum toxin type A for the treatment of lateral canthal rhytides. AB - BACKGROUND: Inactivation of muscles of facial expression by chemodenervation with botulinum toxin remains an off-label indication. Nevertheless, it continues to be a safe and effective technique to improve dynamic rhytides and is the treatment of choice for the hypertrophic lateral fibers of the orbicularis oculi muscle that can cause the superimposed crow's feet. OBJECTIVE: Although infrequent and self-limiting, the complication of unexpected muscle weakness from toxin diffusion or erroneous placement is documented. METHODS: However, injection into the pretarsal portion of the orbicularis oculi muscle resulting in unilateral ocular irritation and diminished tear expression as evidenced by a dry eye and an abnormal Schirmer's test has rarely been reported. Direct injection into the pretarsal fibers of the muscle as opposed to diffusion of the toxin into the muscle fibers or the lacrimal gland was consistent with the onset of action of the toxin and the prolonged duration of the ocular symptoms. RESULTS: Treatment consisted of ocular lubrication until the effects of the toxin dissipated and muscle tone returned. Subsequent treatment did not result in a result in a recurrence of adverse sequelae. CONCLUSIONS: Facial muscles are small, not isolated, and often have fibers that interdigitate. An important factor in the administration of botulinum toxin is the identification of the muscles responsible for the corresponding rhytide. Precise knowledge of muscular anatomy and function will aid in minimizing this and other potential complications. PMID- 11860428 TI - Professional errors caused by lasers and intense pulsed light technology in dermatology and aesthetic medicine: preventive strategies and case studies. AB - BACKGROUND: The use of lasers and intense pulsed light (IPL) technology has become established practice in dermatology and aesthetic medicine. The increase in popularity and widespread use of such equipment has been accompanied by a sharp increase in the number of case reports about professional errors. OBJECTIVE: We present 14 case studies of professional errors. METHODS: Selected representative case reports are used to illustrate and discuss typical professional errors and serve as the basis for creating preventive strategies. RESULTS: Recommendations have been developed for the following areas: physician training, patient information, documentation, diagnosis, indication, test treatment, and performing treatment. CONCLUSION: The use of lasers and IPL technology in dermatology and aesthetic medicine requires practitioners not only to have high levels of training and experience, but also to exercise professional judgment. In spite of all of the precautions taken, the risk of complications and side effects can be reduced but not completely eliminated. PMID- 11860427 TI - Reversible cardiac arrest after polidocanol sclerotherapy of peripheral venous malformation. AB - BACKGROUND: Polidocanol sclerotherapy is a well-established therapeutic modality for the treatment of venous malformations. Systemic complications are extremely rare. OBJECTIVE: To report a case of cardiac complication after polidocanol injection of peripheral venous malformation. METHODS: A case report and a review of the English language literature using a published MEDLINE search strategy. RESULTS: A patient undergoing polidocanol sclerotherapy for a symptomatic venous malformation of the right inferior limb developed cardiac arrest shortly after injection of the sclerosing agent which was promptly reversed. CONCLUSION: Systemic complications following sclerotherapy may occur even when the sclerosant is injected in peripheral veins or venous malformations. Clinicians should be alerted to the possibility of uncommon but life-threatening adverse effects. PMID- 11860430 TI - Botulinum toxin A: successful decollete rejuvenation. AB - BACKGROUND: Decollete wrinkles develop with age. In women more than 35 years of age these wrinkles are still transient, but become permanent at about age 50. In a substantial number of women decollete wrinkles, seem to be associated with an insertion of platysma exceeding the second intercostal space. We report botulinum A toxin therapy of these wrinkles in five patients. OBJECTIVE: To show the effect of botulinum A toxin on decollete wrinkles. METHODS: All five women with decollete lines treated with botulinum A had different varieties of the platysma muscle inserting deep down beneath the fourth intercostal space. During platysma contraction injection points were marked and a certain dosage of toxin was applied. RESULTS: Two weeks after therapy a significant improvement of the treated decollete region was observed. CONCLUSION: These observations indicated that botulinum A toxin can be an effective and safe method in the temporary management of decollete wrinkles. It should therefore be considered as a new adjuvant treatment in cosmetic decollete rejuvenation. PMID- 11860429 TI - Alternative greater saphenous vein-sparing surgery: valvuloplasty combined with axial transposition of a competent tributary vein for the treatment of primary valvular incompetence---18-month follow-up. AB - BACKGROUND: The greater saphenous vein (GSV) is one of the best grafts for vascular reconstruction, and a variety of sparing methods in patients with primary varicose veins have been performed. Of these, valvuloplasty of the subterminal valve is useful, but reflux in the proximal GSV via a competent tributary vein still remains. To minimize the subsequent reflux, we propose a new method, "valvuloplasty combined with axial transposition of a competent tributary vein" for the treatment of GSV incompetence. OBJECTIVE: To compare this new method with single valvuloplasty. METHODS: Seventy-eight limbs in 65 patients with GSV incompetence were included. Of these limbs, 38 underwent angioscopic valvuloplasty of the subterminal valve alone (V group). The remaining 40 were treated by angioscopic valvuloplasty combined with axial transposition of a competent tributary vein (V + T group). A competent tributary vein was identified by duplex scan in the thigh before surgery. After angioscopic valvuloplasty had been done, a competent tributary vein was exposed, and was cut 1.5 cm distal to its insertion. The distal cut end of the vein was then transposed to the GSV and end-to-side anastomosis was performed using 8-0 nylon under magnification. The incompetent GSV was ligated between the insertion and the anastomosed region. Venous hemodynamic changes were analyzed using air plethysmography (APG). Values obtained by9 APG included venous volume (VV), venous filling index (VFI), ejection fraction (EF), and residual venous function (RVF). The follow-up period was 18 months. Intergroup differences were analyzed with the Wilcoxon ranked sum test for nonparametric distribution. RESULTS: There were no statistical differences in age, gender, or clinical presentation between the two groups. In the V group, 27 limbs had reflux in the proximal GSV (67.4%). On the other hand, only 6 limbs (13.3%) showed minor reflux in the V + T group. A significant difference was seen in VFI at 1 year, and a continuous increase was observed in the V group during the follow-up examinations (P =.0035, VFI = 2.50 plus minus 1.21, 1.14 plus minus 0.42 at 18 months, respectively). CONCLUSION: Valvuloplasty combined with axial transposition of a competent tributary vein gives a better result than valvuloplasty alone at the 18-month follow-up. A competent valve in this location can be expected to improve VFI to a normal range. PMID- 11860431 TI - Short-term histologic effects of nonablative resurfacing: results with a dynamically cooled millisecond-domain 1320 nm Nd:YAG laser. AB - BACKGROUND: It is widely believed that nonablative laser techniques can lead to dermal collagen remodeling without the obvious epidermal injury and the wound created with ablative approaches. This occurs when dermal collagen injury is induced without visible injury to the overlying epidermis. OBJECTIVE: To examine the acute histologic effects both 1 hour and several days after standardized treatment protocols of dynamically cooled millisecond domain Nd:YAG 1320 nm laser to provide further insight into the mechanism of action of nonablative resurfacing. METHODS: Multiple adjacent sites on the preauricular area of the cheek of 10 patients were biopsied following one to three laser passes of dynamically cooled millisecond-domain Nd:YAG 1320 nm laser. Biopsies were performed at 1 hour and at 3 days following a single treatment. The number of passes was varied from one to three and Tmax (peak temperature measured by integrated radiometer) during treatment was targeted for 45-48 degree C. RESULTS: At 1 hour after treatment, epidermal spongiosis and edema of the basal cell layer were present in all the specimens treated with three passes. At 3 days the three pass samples also showed microthrombosis, widened vessels, sclerosis of the vessel walls, and infiltration of neurophilic granulocytes. The occurrence of these histologic findings correlated well with the presence of clinical improvement (judged by photographs) at 8 weeks after treatment. Acute histologic changes and clinical improvement were not observed below treatment temperatures of Tmax 45 degree C or after one pass alone. Repeated temperatures above a Tmax of 48 degree C incurred risk of epidermal injury. CONCLUSION: Even though longer term histologic findings have confirmed the collagen synthesis component of 1320 nm Nd:YAG laser, our data indicate that there may be some additional factors other than dermal collagen heating with subsequent collagen repair. The concept of true "nonablative resurfacing" may involve some form of subclinical epidermal injury that improves the clinical outcome. Acute changes involving superficial blood vessel injury with cytokine release may also be implicated. Our histologic findings suggest that three passes with fluence and cooling adjusted to a Tmax of 45 degree C-48 degree C yields improved clinical results. PMID- 11860432 TI - Multiple color changes following laser therapy of cosmetic tattoos. AB - OBJECTIVE: To emphasize the wisdom of small test areas when treating cosmetic tattoos and the need for multiple laser systems. METHODS: A 48-year-old woman requested removal of permanent makeup (cosmetic tattoos) of her eyebrows and around her lips. Physical examination revealed a brown tattoo of both eyebrows and dark red lip liner around both lips. A test area was performed on the red tattoo of the lips. A frequency-doubled Nd:YAG laser (532 nm, 2.0 J/cm2, 2 mm spot size) was used for the lip area, while the same laser at 1064 nm, 3.9 J cm2, 2 mm spot size was utilized for the eyebrows. The lip area immediately turned black. The patient returned for follow-up 1 month later; the black ink on the lip was treated with the same laser at 1064 nm, 3 mm spot size, 4.2 J/cm2, with satisfactory resolution in two monthly treatments. Both brown eyebrow turned bright orange and were treated with 532 nm, 3 mm, 3.0 J/cm2. One month later the eyebrows were a mixture of yellow ink and dark green. The yellow area was treated with 532 nm, 3 mm, 2.3 J/cm2, while the dark green was treated with the 1064 nm, 3 mm spot size, 4.2 J/cm2. One month later little improvement was noted, so Q switched ruby laser at 694 nm, 6 mm spot size, 16 J/cm2 was utilized. An additional four monthly treatments were given utilizing a combination of both ruby and 532 nm ND:YAG lasers for green and yellow pigment, respectively. RESULTS: Significant but incomplete resolution of the tattoo ink was achieved. CONCLUSION: Multiple laser systems are needed to remove cosmetic tattoos. Test areas must be done before treatment. PMID- 11860433 TI - Eleven pearls for cosmetic earlobe repair. AB - Earlobe repair is a common request in cosmetic facial surgery. Earlobe tears result from a variety of traumatic situations. A resurgence in body piercing and multiple facial and ear piercings is occurring in college-aged individuals. This increase in piercing will bring more piercing-related complications to the cosmetic surgeon. The author has outlined 11 surgical pearls to provide improved cosmetic results in the repair of torn earlobes. Also described is immediate repiercing of the earlobe at the time of the surgical repair. Preventive practices to decrease earring and earlobe trauma are also reviewed. PMID- 11860434 TI - A case of editorial mohs surgery: we just need to cut a little more. PMID- 11860435 TI - Cyclopamine as a potential therapeutic agent for treatment of tumors related to hedgehog pathway mutations. PMID- 11860436 TI - The criss-cross tie-over tacking suture. PMID- 11860437 TI - Nasalis myocutaneous island pedicle flap with bilevel undermining for repair of lateral nasal defects. AB - BACKGROUND: The superiorly based nasalis myocutaneous island pedicle flap with bilevel undermining is a recently described bipedicled flap used for repair of nasal tip and supratip defects. This flap was used for the repair of small to medium-size defects of the nasal tip and supratip regions. OBJECTIVE: To describe a single-pedicled modification of this flap for the reconstruction of lateral nasal defects at or above the alar groove. METHODS: With the use of diagrams and fresh cadaver dissection, we demonstrate the method of preparing, raising, and placement of this new flap. We further elucidate in an illustrative way the efficacy of bilevel undermining as it is used for achieving greater flap mobility. Finally, we show representative cases with pre- and postoperative results. RESULTS: We describe the senior author's 4-year experience in the development of this flap. CONCLUSION: The nasalis myocutaneous island pedicle flap with bilevel undermining, a new method for repair of lateral nasal defects at or above the alar groove, yields excellent functional and cosmetic results. Bilevel undermining is a new and valuable method of achieving greater tissue mobility in muscle-based flaps. PMID- 11860438 TI - Repair of a defect of the lower lip. PMID- 11860439 TI - Inexpensive liquid nitrogen container. PMID- 11860440 TI - Reducing pain during hair transplant lidocaine infiltration. PMID- 11860441 TI - Not the presence but the amount of clonal DNA detectable in remission of acute myeloid leukemia is predictive for relapse. AB - OBJECTIVES: The persistence of clonal cells after chemotherapy, or a re-emerging of clonal cells in remission (CR) or at relapse in patients with acute myeloid leukemia (AML) was studied to assess the prognostic significance of the amount of clonal DNA in predicting the clinical outcome. METHODS: Clonal rearrangements in the gene sequences of retinoic acid receptor (RAR) alpha, major breakpoint cluster region (M-bcr), immunoglobulin (Ig)-JH, T-cell receptor (TcR) beta, myeloid lymphoid leukemia or cytokines (GM-CSF, G-CSF, IL-3) detected in bone marrow samples from 37 patients with primary AML (pAML) or secondary AML (sAML) were investigated. A relative increase or decrease of clonal DNA in the course of AML was evaluated by comparing the optical densities of DNA bands of the rearranged genes and the total amount of DNA. RESULTS: High amounts of clonal DNA were detectable at diagnosis, during persisting disease and at relapse (O 39%, 35%, or 38% of total DNA, respectively), compared to 20% in complete remission (CR). Amounts of clonal DNA (except for Ig-JH gene rearrangements) were of prognostic significance at diagnosis, patients with less than 33% clonal DNA were characterized by significantly longer relapse-free survival times (all cases: p = 0.01; pAML: p = 0.002). Patients in CR exhibiting less than 5% (all cases) or 15% (pAML) clonal DNA showed longer relapse-free survival times (p = 0.08 or p = 0.03, respectively). Vice versa, significantly higher amounts of clonal DNA (all cases 51% vs. pAML 54%) could be detected in cases studied at diagnosis who relapsed in the following 5 months (all cases p = 0.01) or 14 months (pAML p = 0.007). Significantly higher amounts of clonal DNA (33%) could be detected in cases studied in CR who relapsed in the following 4 months (all cases p = 0.002 or pAML p = 0.006, respectively). Moreover, we could prove disease progression on a cellular level months before the clinical onset of sAML after a period of MDS. CONCLUSIONS: Clonal, gene-rearranged DNA is regularly detectable at diagnosis and during persisting AML, in CR and at relapse. However, the presence, rather than the amount of clonal DNA detectable in CR is predictive for relapse. These data might indicate the significance of gene rearrangement analyses in the course of AML to identify cases with a high risk of relapse, independently from the karyotype. PMID- 11860443 TI - Long-term administration of high-dose deferoxamine 2 days per week in thalassemic patients. AB - OBJECTIVE: Compliance with parenteral administration of deferoxamine is often poor in thalassemic patients with iron overload. We tested the efficacy and tolerance of the drug at high dosage 2 d per week for 24 months in two adult thalassemic patients with permanently high serum ferritin using a portable pump and an implanted chamber. METHODS: Deferoxamine was administered using a pressure operated portable pump through an implanted chamber. The patients were infused over 48 h every week with 198 mg/kg/d (patient 1) and 170 mg/kg/d (patient 2). Serum ferritin levels were measured at regular intervals. RESULTS: Serum ferritin decreased progressively from 2967 to 457 microg/L in patient 1 and from 6476 to 1951 microg/L in patient 2. Compliance and tolerance to treatment were excellent in the two patients. CONCLUSION: Intravenous administration of high-dose deferoxamine over 48 h per week using a portable pump and implanted chamber improved compliance in two thalassemic adult patients, resulting in a significant decrease in iron overload. We suggest that high-dose chelation therapy should be assessed in selected groups of iron-overload thalassemic patients receiving regular blood transfusions. PMID- 11860442 TI - FAB M4 and high CD14 surface expression is associated with high cellular resistance to Ara-C and daunorubicin: implications for clinical outcome in acute myeloid leukaemia. AB - In 145 adult patients diagnosed with non-M3 acute myeloid leukaemia (AML) the relevance of FAB-subtype and immunophenotype to in vitro cellular drug resistance towards the anthracyclines aclarubicin (Acla) and daunorubicin (Dau), and the nucleoside analogue cytarabine (Ara-C), as well as other antileukaemic drugs, was investigated using a 4-d MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide) assay. We demonstrate that high CD14 expression is highly significantly associated with high cellular Ara-C and Dau resistance in univariate as well as multivariate analyses. FAB subtypes with highest and lowest cellular Ara-C resistance were M4 and M5, respectively (P < 0.01, one-way anova), whereas FAB subtypes with highest and lowest cellular Dau resistance were M4 and M1, respectively (P < 0.01, one-way anova). By contrast, no significant differences in cellular drug resistance towards Acla could be demonstrated among FAB subtypes. Furthermore, in two cohorts of AML patients treated by two different regimens for remission induction over a period of 15 yr (1985-94, n = 159 and 1995-99, n = 76, respectively) we demonstrate in univariate analyses a significance of CD14 expression with respect to clinical outcome. With the exception of significance to probability of obtaining complete remission in the first cohort (P = 0.03, logistic regression), this significance was, however, lost in multivariate analyses. It was demonstrated that FAB-M4 patients were older than M5 patients and that high CD14 expression was associated with the presence of secondary AML and older age. We conclude that although cases with high blast cell CD14 expression (and FAB-M4 cases) were more resistant to Ara-C as well as Dau in vitro, the clinical and biological significance of this may be debatable because of interactions with major prognostic factors in AML. PMID- 11860444 TI - Lack of pathogenic mutations in the 5'-untranslated region of the thrombopoietin gene in patients with non-familial essential thrombocythaemia. AB - Thrombopoietin (TPO) is thought to be the major physiological regulator of thrombopoiesis, and, in general, circulating levels are inversely proportional to megakaryocyte and platelet mass. However, normal or elevated TPO levels are found in patients with essential thrombocythaemia (ET) and the reason for this is not fully understood. Recent studies have shown that four kindreds with hereditary thrombocythaemia (HT) have point mutations in the 5'-untranslated region (UTR) of the TPO gene which lead to increased TPO translation. In order to determine whether similar mutations are present in apparently acquired ET, in particular in those patients with polyclonal myelopoiesis, we have studied this region in 50 ET patients using neutrophil DNA. The known HT mutations were investigated using polymerase chain reaction with mismatch primers and restriction enzyme digestion; only wild-type alleles were detected. Single-stranded conformation polymorphism (SSCP) analysis of exons 1-4 identified a C-->T substitution at nucleotide 3767. However, this appears to be a common polymorphism, as it was present at the same frequency in haematologically normal controls and is unlikely to be of pathological significance. These results demonstrate that mutations in the 5' UTR of the TPO gene are not the cause of the normal or elevated TPO levels in acquired ET. PMID- 11860445 TI - Prevalence of retinopathy in patients with anemia or thrombocytopenia. AB - BACKGROUND AND OBJECTIVES: Fundus lesions can be the accompanying symptom in many hematological diseases. In cases of anemia or thrombocytopenia, the exact mechanism leading to fundus abnormalities is not completely understood. The aim of this study was to calculate the prevalence of fundus lesions in anemic and thrombocytopenic patients, and define the risk factors for retinopathy. DESIGN AND METHODS: This cross-sectional study involved 226 patients with anemia and/or thrombocytopenia and a control group of 47 age-matched subjects. The studied variables were age, gender, hemoglobin levels, mean cell volume (MCV), red cell distribution width coefficient variation (RDW-CV), the type of onset of anemia, platelet counts, mean platelet volume (MPV), platelet distribution width (PDW), the platelet-large-cell ratio (P-LCR), prothrombin time (PT) and partial thromboplastin time (PTT). RESULTS: Retinopathy was observed in 28.3% of the patients as a whole, the presence of fundus lesions being closely associated with severe anemia (Hb < 8 g/dL) and severe thrombocytopenia (PLT < 50 x 109/L). Among the patients with concomitant anemia and thrombocytopenia, the incidence of retinopathy was 38%. Age, low hemoglobin levels, platelet counts, RDW-CV, and increased MCV, MPV and P-LCR were all significantly associated with the presence of fundus lesions at univariate analysis. INTERPRETATION AND CONCLUSIONS: Retinopathy is a frequent finding in anemic and thrombocytopenic patients, although it is often not significant clinically. As retinal hemorrhages were found in all of the patients with concomitant severe anemia and thrombocytopenia, it is recommended that all such patients undergo routine fundus examinations. PMID- 11860447 TI - Benign thymic hyperplasia after chemotherapy for acute myeloid leukemia. AB - Thymic hyperplasia can occur after cytotoxic therapy for various malignancies. The possible cause could be rebound enlargement after initial atrophy caused by these drugs. During the treatment of hematological malignancies this could be a cause of great concern. We report here a case of thymic hyperplasia after chemotherapy for acute myeloid leukemia. Awareness of this unusual side-effect may prevent needless investigation and therapy. PMID- 11860446 TI - Assessment of bone marrow stem cell reserve and function and stromal cell function in patients with severe congenital neutropenia. AB - OBJECTIVE: To investigate further the cellular defect responsible for impaired granulopoiesis in severe congenital neutropenia (SCN), we have evaluated bone marrow (BM) stem cell reserve and function and BM stromal cell myelopoiesis supporting capacity in two patients with SCN. METHODS: BM primitive stem cells and myeloid progenitor cells were assessed using flow cytometry, limiting dilution assay, clonogenic assays, and long-term BM cultures (LTBMC). BM stroma function was assessed by evaluating the ability of irradiated stromal layers from the patients to induce granulocyte-macrophage colony formation (CFU-GM) by normal CD34+ cells. RESULTS: Compared to the normal controls (n = 37), SCN patients displayed a low percentage of CD34+/CD38+ cells (P < 0.05), low CFU-GM colony formation by highly purified CD34+ cells (P < 0.05), low CFU-GM recovery in LTBMC (P < 0.05), and normal primitive stem cells as indicated by the frequency of CD34+/CD38- cells and the number of long-term culture initiating cells. Patient BM stromal layers exhibited normal myelopoiesis supporting capacity as shown by the CFU-GM content of irradiated LTBMC recharged with normal CD34+ cells. In addition, patient LTBMC supernatants displayed 20-fold normal granulocyte colony stimulating factor and 2-fold normal granulocyte-macrophage colony stimulating factor levels. CONCLUSION: These data show that primitive BM stem cells and stromal cells are not affected in SCN patients, while they support further the concept of a primary defect at the myeloid progenitor cell level. To know the differentiation stage at which the underlying defect causes the malfunction will be relevant for further elucidation of its nature at the molecular level. PMID- 11860448 TI - Successful treatment with low-dose splenic irradiation for massive splenomegaly in an elderly patient with hairy-cell leukemia. AB - The therapeutic approach to hairy-cell leukemia (HCL) is in some instances still debated. Although management with alpha-interferon and purine analogues is well established, there is an alternative role for therapeutic splenectomy in patients with massive splenomegaly who have failed to respond to systemic therapy. Most patients with HCL will not be suitable for treatment with splenectomy as their ages at diagnosis are high. Here, we report an elderly Japanese HCL patient whose refractory massive splenomegaly responded well to low-dose splenic irradiation. PMID- 11860449 TI - Molecular characterization of (deltabeta)(0)/beta(0)-thalassemia and (deltabeta)(0)-thalassemia/hemoglobin E in Thai patients. AB - Two cases of the Thai thalassemia patients with compound heterozygosities for (deltabeta)(0)/beta(0)-thalassemia and (deltabeta)(0)-thalassemia/hemoglobin E have been reported. The first case was a 8-yr-old boy who had the following hematologic data: Hb 6.5 g/dL, Hct 20.5%, MCV 70.4 fL, MCH 22.3 pg and MCHC 31.7 g/dL. Hemoglobin analysis revealed 1.9% hemoglobin A2 and 91.7% hemoglobin F. The second case, with Hb 13.9 g/dL, Hct 41.5%, MCV 69.5 fL, MCH 22.5 pg and MCHC 32.2 g/dL, was a 16-yr-old male who had 46.1% hemoglobin E and 49.8% hemoglobin F. Globin gene analyses showed that both probands carried the same deletional type (deltabeta)(0)-thalassemia trans to the 4 bp deletions in codons 41/42 beta(0) thalassemia and to the betaE-globin gene, respectively. Polymerase chain reaction and DNA sequence analyses demonstrated that the 5' breakpoint of the (deltabeta)(0)-thalassemia deletion was located in the second intron of the delta globin gene and that the 3' breakpoint lay within a cluster of LI repetitive sequences at 4.7 kb 3' to the beta-globin gene. PMID- 11860450 TI - Hemochromatosis in a Lithuanian with HFE C282Y homozygosity and C282Y allele frequencies in the Baltic Sea region. PMID- 11860451 TI - Plasma thrombopoietin levels in Sebastian platelet syndrome. PMID- 11860452 TI - Platelet-derived growth factor (PDGF) in human acute myelogenous leukemia: PDGF receptor expression, endogenous PDGF release and responsiveness to exogenous PDGF isoforms by in vitro cultured acute myelogenous leukemia blasts. AB - We investigated effects of Platelet-derived growth factor (PDGF) and Platelet factor 4 (PF-4) on the functional characteristics of native, human acute myelogenous leukemia (AML) blasts. AML blast expression of the PDGF-receptor alpha-chain was detected for a subset of patients (45%), whereas PDGF-receptor beta-chain expression was detected for most patients (90%). Constitutive AML blast release of the PDGF-AB isoform (the major form also derived from normal platelets) was detected for 43% of patients, whereas PDGF-BB release was not detected for any patient. The PDGF isoforms AA, AB and BB had dose-dependent and divergent effects on spontaneous and cytokine-dependent AML blast proliferation, whereas for constitutive cytokine secretion (IL-1beta, IL-6, TNF-alpha) inhibitory effects were rare and all three isoforms usually had no effect or enhanced the constitutive secretion. The PDGF effects were caused by a direct effect on the AML blasts and were not dependent on the presence of serum. The PDGF effects could also be detected after in vitro culture of AML cells in the presence of IL-4+ granulocyte-macrophage colony stimulating factor. PF-4 had divergent effects on proliferation and cytokine secretion by native AML blasts. Our results suggest that exogenous (e.g. platelet-secreted) PDGF and PF-4 can function as regulators of leukemic hematopoiesis and possibly also modulate the function of residual AML cells in peripheral blood stem cell grafts. On the other hand, endogenous release of PDGF-AB by native blasts may modulate the function of normal cells in the bone marrow microenvironment (e.g. bone marrow stromal cells). PMID- 11860454 TI - Characterization of the single-channel properties of NMDA receptors in laminae I and II of the dorsal horn of neonatal rat spinal cord. AB - The single-channel properties of native NMDA receptors in laminae I and II of the dorsal horn of the neonatal rat spinal cord were studied using outside-out patch clamp techniques. These receptors were found to have several features that distinguish them from native NMDA receptors elsewhere in the CNS. Single-channel currents activated by NMDA (100 nm) and glycine (10 microm) exhibited five distinct amplitude components with slope-conductance values of 19.9 +/- 0.8, 32.9 +/- 0.6, 42.2 +/- 1.1, 53.0 +/- 1.0 and 68.7 +/- 1.5 pS. Direct transitions were observed between all conductance levels but transitions between 69-pS openings and 20-, 33- and 42-pS openings were rare. There was no significant difference in the frequency of direct transitions from 42- to 20-pS compared to 20- to 42-pS transitions. The Kb (0 mV) for Mg2+ was 89 microm. The Mg2+ unblocking rate constant was similar to other reported values. However, the Mg2+ blocking rate constant was larger than other reported values, suggesting an unusually high sensitivity to Mg2+. The NR2B subunit-selective antagonist, ifenprodil, had no significant effect on overall channel activity but significantly decreased the mean open time of 53-pS openings. These results suggest neonatal laminae I and II NMDA receptors are not simply composed of NR1 and NR2B subunits or NR1 and NR2D subunits. It is possible that these properties are due to an as yet uninvestigated combination of two NR2 subunits with the NR1 subunit or a combination of NR3A, NR2 and NR1 subunits. PMID- 11860453 TI - Differential sensitivity of medium- and large-sized striatal neurons to NMDA but not kainate receptor activation in the rat. AB - Infrared videomicroscopy and differential interference contrast optics were used to identify medium- and large-sized neurons in striatal slices from young rats. Whole-cell patch-clamp recordings were obtained to compare membrane currents evoked by application of N-methyl-d-aspartate (NMDA) and kainate. Inward currents and current densities induced by NMDA were significantly smaller in large- than in medium-sized striatal neurons. The negative slope conductance for NMDA currents was greater in medium- than in large-sized neurons and more depolarization was required to remove the Mg2+ blockade. In contrast, currents induced by kainate were significantly greater in large-sized neurons whilst current densities were approximately equal in both cell types. Spontaneous excitatory postsynaptic currents occurred frequently in medium-sized neurons but were relatively infrequent in large-sized neurons. Excitatory postsynaptic currents evoked by electrical stimulation were smaller in large- than in medium sized neurons. A final set of experiments assessed a functional consequence of the differential sensitivity of medium- and large-sized neurons to NMDA. Cell swelling was used to examine changes in somatic area in both neuronal types after prolonged application of NMDA or kainate. NMDA produced a time-dependent increase in somatic area in medium-sized neurons whilst it produced only minimal changes in large interneurons. In contrast, application of kainate produced significant swelling in both medium- and large-sized cells. We hypothesize that reduced sensitivity to NMDA may be due to variations in receptor subunit composition and/or the relative density of receptors in the two cell types. These findings help define the conditions that put neurons at risk for excitotoxic damage in neurological disorders. PMID- 11860455 TI - Neither major depression nor glucocorticoid treatment affects the cellular integrity of the human hippocampus. AB - In major depression, decreased hippocampal volume has been attributed to hypercortisolemia, a frequent sign of the disorder, because in animals an excess of corticosteroids has led to dendritic atrophy, astrogliosis and loss of neurons in this brain region. The present study is the first to investigate the structural integrity of the human hippocampus in major depression and following glucocorticoid treatment. Post-mortem hippocampal tissue from 15 patients who had had major depression or bipolar affective disorder, 10 patients who had been treated with glucocorticoids and 16 controls was assessed using haematoxylin eosin, Nissl and Bodian staining. The patterns of reactive astrogliosis (glial fibrillary acidic protein, GFAP), synaptic density (synaptophysin), synaptic reorganization (growth-associated protein B-50) and early signs of Alzheimer's disease (Alz-50) were examined immunocytochemically. Multivariate analysis, with the patients' age, tissue fixation time and postmortem delay as covariates, was performed. There was no evidence of neuronal cell loss or other major morphological alterations in any of the groups, nor was there a significant change in the distribution pattern of synaptophysin or Alz-50. Changes in B-50 and GFAP staining were observed in the steroid-treated and depressed patients in areas CA1 and CA2 only. The human hippocampus in major depression and after glucocorticoid treatment does not reveal any major morphological changes or signs of neuronal cell death, but does show subtle alterations in B-50 and GFAP expression in selected parts of the pyramidal cell layer. PMID- 11860456 TI - Developmental expression of the calcium release channels during early neurogenesis of the mouse cerebral cortex. AB - The developmental changes of intracellular calcium release channels of mouse neocortex were studied at the onset of neurogenesis, which occurs between embryonic days E11 and E17. The three main isoforms of the two families of intracellular calcium release channels, namely the inositol trisphosphate receptors (IP3R) and the ryanodine receptors (RyR), were detected by their transcripts in the cerebral hemispheres, as early as stage E11. The major isoforms of each family, IP3R-1 and RyR-2, were found at the protein level by Western blot analysis. Expression of these proteins increases progressively throughout brain development. Their localization in coronal sections of cortex has been observed by immunodetection from E12, and compared to the TuJ1 (anti class III beta-tubulin antibody) neuronal specific labelling. The expression of both channels is greatly enhanced after E12, and both were seen to be present in most of the proliferative and neuronal cells of the slice. Between E12 and E13, there is a striking transition in the pattern of calcium release elicited by specific agonists of these channels, thimerosal for IP3R and caffeine for RyR. The signals induced by thimerosal were not zone-specific, while the observed calcium release signals induced by caffeine were predominantly restricted out of the ventricular zone. This zone-specific caffeine sensitivity is consistent with the main RyR localization immunodetected at E13. Our results indicate that there is a time lag of several days between the molecular detection of calcium release channels and their functional expression, around the time of neuronal differentiation. Altogether, they provide a molecular basis for analyzing the developmental modulation of calcium signals useful for neurogenesis progression. PMID- 11860457 TI - Brain targeting and glomerulus formation of two olfactory neuron populations expressing related receptor types. AB - Olfactory sensory neurons expressing different members of the mOR37 odourant receptor subfamily send their axons to distinct glomeruli located in the immediate vicinity in the olfactory bulb [Strotmann, J., Conzelmann, S., Beck, A., Feinstein, P., Breer, H. & Mombaerts, P. (2000) J. Neurosci., 20, 6927-6938]. In this study, the potential of transgenic mouse lines was used to explore the onset of receptor expression, the outgrowth of axons as well as the glomerulus formation for two neuron populations expressing different mOR37 subtypes. The data indicate a synchronous time course of these features for both neuron populations. From E15 until the day of birth, the axons of the two mOR37 populations terminate in a common, small area of the presumptive olfactory bulb. During a short postnatal phase, the two axon populations segregate into distinct, protoglomerular structures; some aberrant fibers can still be observed during this period. PMID- 11860458 TI - Organization of inputs from cingulate motor areas to basal ganglia in macaque monkey. AB - The cingulate motor areas reside within regions lining the cingulate sulcus and are divided into rostral and caudal parts. Recent studies suggest that the rostral and caudal cingulate motor areas participate in distinct aspects of motor function: the former plays a role in higher-order cognitive control of movements, whereas the latter is more directly involved in their execution. Here, we investigated the organization of cingulate motor areas inputs to the basal ganglia in the macaque monkey. Identified forelimb representations of the rostral and caudal cingulate motor areas were injected with different anterograde tracers and the distribution patterns of labelled terminals were analysed in the striatum and the subthalamic nucleus. Corticostriatal inputs from the rostral and caudal cingulate motor areas were located within the rostral striatum, with the highest density in the striatal cell bridges and the ventrolateral portions of the putamen, respectively. There was no substantial overlap between these input zones. Similarly, a certain segregation of input zones from the rostral and caudal cingulate motor areas occurred along the mediolateral axis of the subthalamic nucleus. It has also been revealed that corticostriatal and corticosubthalamic input zones from the rostral cingulate motor area considerably overlapped those from the presupplementary motor area, while the input zones from the caudal cingulate motor area displayed a large overlap with those from the primary motor cortex. The present results indicate that a parallel design underlies motor information processing in the cortico-basal ganglia loop derived from the rostral and caudal cingulate motor areas. PMID- 11860460 TI - Putative glycine receptors in Hydra: a biochemical and behavioural study. AB - Glycine acts as an inhibitory transmitter in the lower brain stem and spinal cord of vertebrate species, while very few data are yet available to support a similar role in invertebrate nervous systems. Here we report the identification and characterization of glycine receptors in the freshwater polyp Hydra vulgaris (Cnidaria, Hydrozoa) by biochemical and behavioural studies. Saturation experiments revealed the occurrence of one population of binding sites of nanomolar affinity (KD = 33 nm) and low capacity (Bmax = 79 fmol/mg protein) for [(3)H]strychnine. The addition of glycine or taurine (0.1 microm-1 mm) produced a dose-dependent inhibition of [(3)H]strychnine binding. Beta-alanine (0.1-1 mm) did not significantly affect [(3)H]strychnine binding. The pharmacological properties of these receptors compare with those of vertebrate glycine receptors. Stimulation of Hydra polyps by reduced glutathione resulted in a significant increase in the duration of mouth opening in the presence of glycine, taurine or beta-alanine. The enhancement of the response was related both to amino acid (10 100 microm) and to glutathione concentration (1-10 microm). The effects of glycine or its agonists were suppressed by strychnine (1-10 microm). D-serine, a glycine agonist at the vertebrate NMDA receptor, produced opposite effects to those of glycine. The effects of d-serine were suppressed by 5,7 dichlorokynurenic acid but not by strychnine. In vitro, [(3)H]strychnine binding was not displaced by d-serine. These results indicate a dual action of glycine in Hydra tissues. The hypothesis that NMDA receptors may also be present in this elementary nervous system is proposed. PMID- 11860459 TI - Immune surveillance of mouse brain perivascular spaces by blood-borne macrophages. AB - Virchow-Robin's perivascular spaces lie between the basement membrane around pericytes and the basement membrane at the surface of the glia limitans of the brain vessels. They are directly connected to the subpial space and harbour a population of cells distinct from pericytes, perivascular microglia and other cells within perivascular spaces (e.g. T cells and mast cells) in their ability to quickly phagocytose particles from the cerebrospinal fluid (CSF). Morphology, function, and cell surface proteins of these perivascular cells suggest an origin from the monocyte/macrophage lineage. It is currently unclear to what extent these brain perivascular cells represent a resident population of histiocytes or undergo continuous supplementation from blood monocytes. Using transplants of green-fluorescent-protein (GFP)-transfected bone marrow cells, we therefore investigated the replacement of perivascular cells by blood-borne macrophages in adult mice. GFP-positive cells in the perivascular spaces were found as early as 2 weeks post transplantation. The substitution of host perivascular cells by donor-derived macrophages was then evaluated using immunocytochemistry and intraventricular injection of hydrophilic rhodamine-fluorescent tracers. Such tracers diffuse along perivascular spaces and are subsequently phagocytosed by perivascular cells leading to stable phagocytosis-dependent labelling. Thus, the population of newly immigrated macrophages could be related to the total number of perivascular macrophages. This approach revealed a continuous increase of donor-derived perivascular cells. At 14 weeks post transplantation, all perivascular cells were donor-derived. These data show that brain perivascular cells are a population of migratory macrophages and not resident histiocytes. PMID- 11860461 TI - Reduction in CNS scar formation without concomitant increase in axon regeneration following treatment of adult rat brain with a combination of antibodies to TGFbeta1 and beta2. AB - In this study we investigated whether CNS axons regenerate following attenuation of scar formation using a combination of antibodies against two isoforms of transforming growth factor beta (TGFbeta). Anaesthetized adult rats were given unilateral mechanical lesions of the nigrostriatal tract. Implantation of transcranial cannulae allowed wounds to be treated with a combination of antibodies against TGFbeta1 and TGFbeta2 once daily for 10 days postaxotomy. Eleven days post-transection brains from animals under terminal anaesthesia were recovered for histological evaluation. Gliosis, inflammation and the response of dopaminergic nigral axons were assessed by immunolabelling. Treatment with antibodies against TGFbeta1 and TGFbeta2 attenuated (but did not abolish) the response of glial fibrillary acidic protein (GFAP)-immunoreactive astrocytes and of NG2-immunoreactive glia but did not attenuate the response of CR3 immunoreactive microglia and macrophages. However, this reduction in scar formation was not accompanied by growth of cut dopaminergic nigral axons. We conclude that treatment of injured adult rat brain with a combination of antibodies against TGFbeta1 and TGFbeta2 results in a reduction of scar formation but that this is not sufficient to enhance spontaneous long distance CNS axon regeneration. PMID- 11860463 TI - Motion processing in the auditory cortex of the rufous horseshoe bat: role of GABAergic inhibition. AB - This study examined the influence of inhibition on motion-direction-sensitive responses of neurons in the dorsal fields of auditory cortex of the rufous horseshoe bat. Responses to auditory apparent motion stimuli were recorded extracellularly from neurons while microiontophoretically applying gamma aminobutyric acid (GABA) and the GABAA receptor antagonist bicuculline methiodide (BMI). Neurons could respond with a directional preference exhibiting stronger responses to one direction of motion or a shift of receptive field (RF) borders depending on direction of motion. BMI influenced the motion direction sensitivity of 53% of neurons. In 21% of neurons the motion-direction sensitivity was decreased by BMI by decreasing either directional preference or RF shift. In neurons with a directional preference, BMI increased the spike number for the preferred direction by a similar amount as for the nonpreferred direction. Thus, inhibition was not direction specific. BMI increased motion-direction sensitivity by either increasing directional preference or magnitude of RF shifts in 22% of neurons. Ten percent of neurons changed their response from a RF shift to a directional preference under BMI. In these neurons, the observed effects could often be better explained by adaptation of excitation rather than inhibition. The results suggest, that adaptation of excitation, as well as cortex specific GABAergic inhibition, contribute to motion-direction sensitivity in the auditory cortex of the rufous horseshoe bat. PMID- 11860462 TI - The alpha1I T-type calcium channel exhibits faster gating properties when overexpressed in neuroblastoma/glioma NG 108-15 cells. AB - The recently cloned T-type calcium channel alpha1I (Cav3.3) displays atypically slow kinetics when compared to native T-channels. Possible explanations might involve alternative splicing of the alpha1I subunit, or the use of expression systems that do not provide a suitable environment (auxiliary subunit, phosphorylation, glycosylation...). In this study, two human alpha1I splice variants, the alpha1I-a and alpha1I-b isoforms that harbour distinct carboxy terminal regions were studied using various expression systems. As the localization of the alpha1I subunit is primarily restricted to neuronal tissues, its functional expression was conducted in the neuroblastoma/glioma cell line NG 108-15, and the results compared to those obtained in HEK-293 cells and Xenopus oocytes. In Xenopus oocytes, both isoforms exhibited very slow current kinetics compared to those obtained in HEK-293 cells, but the alpha1I-b isoform generated faster currents than the alpha1I-a isoform. Both activation and inactivation kinetics of alpha1I currents were significantly faster in NG 108-15 cells, while deactivating tail currents were two times slower, compared to those obtained in HEK-293 cells. Moreover, the alpha1-b isoform showed significantly slower deactivation kinetics both in NG 1080-15 and in HEK-293 cells. Altogether, these data emphasize the advantage of combining several expression systems to reveal subtle differences in channel properties and further indicate that the major functional differences between both human alpha1I isoforms are related to current kinetics. More importantly, these data suggest that the expression of the alpha1I subunit in neuronal cells contributes to the "normalization" of current kinetics to the more classical, fast-gated T-type Ca2+ current. PMID- 11860464 TI - C-type natriuretic peptide (CNP) regulates cocaine-induced dopamine increase and immediate early gene expression in rat brain. AB - The neuropeptide C-type natriuretic peptide (CNP) is the primary biologically active natriuretic peptide in brain. Using in situ hybridization, the present report demonstrates that CNP regulates egr-1, c-fos and junB immediate early gene expression in rat brain. In the frontal cortex, CNP induced immediate early gene expression whereas it inhibited dose-dependently the cocaine-induced early gene expression in the dopaminergic projection fields nucleus accumbens and caudate putamen. CNP may produce its effect directly on dopaminergic neurons because we found that its receptor, guanylyl cyclase GC-B, was expressed in the mesencephalon where dopaminergic neurons originate, as well as in their projection fields. The inhibition by CNP of the early gene expression elicited by cocaine in the caudate-putamen is correlated with a CNP-evoked decrease in cocaine-induced rise in extracellular dopamine, measured by in vivo microdialysis experiments. The significance of the inhibition of cocaine-induced dopamine release and early gene induction by the endogenous peptide CNP is demonstrated by data indicating that CNP reduced the cocaine-induced spontaneous locomotor activation. By inhibiting dopaminergic neuronal activity, CNP represents a potential negative regulator of related behavioural effects of cocaine. PMID- 11860465 TI - Discharge patterns of neurons in the medial pontobulbar reticular formation during fictive mastication in the rabbit. AB - In this study, we describe functional characteristics of neurons forming networks generating oral ingestive motor behaviours. Neurons in medial reticular nuclei on the right side of the brainstem between the trigeminal and hypoglossal motor nuclei were recorded in anaesthetized and paralysed rabbits during two types of masticatory-like motor patterns induced by electrical stimulation of the left (contralateral) or right (ipsilateral) cortical masticatory areas. Sixty-seven neurons in nucleus reticularis pontis caudalis (nPontc), nucleus reticularis parvocellularis (nParv), and nucleus reticularis gigantocellularis (Rgc) were studied. These were classified as phasic or tonic depending on their firing pattern during the fictive jaw movement cycle. Phasic neurons located in the dorsal part of nPontc were active during the jaw opening phase, whilst those in dorsal nParv tended to fire during the closing phase. In most neurons, burst duration and firing frequency changed between the two motor patterns, but there was little change in phase of firing. Tonic units were mainly recorded in the ventral half of nPontc, and at the junction between Rgc and caudal nParv. Cortical inputs with short latency from the contralateral masticatory area were more frequent in phasic (82%) than tonic (44%) neurons, whilst inputs from the ipsilateral cortex were equal in the two subgroups (57% and 56%). Phasic neurons had significantly shorter mean contralateral than ipsilateral cortical latencies, whilst there was no difference among tonic neurons. Intra- and perioral primary afferent inputs activated both types of neurons at oligo-synaptic latencies. Our results show that subpopulations of neurons in medial reticular nuclei extending from the caudal part of the trigeminal motor nucleus to the rostral third of the hypoglossal motor nucleus are active during the fictive masticatory motor behaviour. Unlike masticatory neurons in the lateral tegmentum, the medial subpopulations are spatially organized according to discharge pattern. PMID- 11860466 TI - Change blindness and time to consciousness. AB - Detection of changes in a visual scene can be substantially delayed when the original and the modified image are separated by a brief screen flicker. We used this phenomenon of "change blindness" to find when the brain detects the mismatch in relation to when the observer reports it, and whether changes in identity and position are processed similarly. Event-related brain potentials (ERPs) recorded while the subjects searched for the change in alternating series of images showed that the epoch during which they indicated detection was characterized by a marked positivity from 300 to 700 ms. Analysis of data from image presentations preceding the subjects' response revealed a similar but smaller ERP positivity one (identity) or even two (position) epochs before detection. As each epoch lasted 1500 ms, the brain may register a change as early as 3000 ms before the observer. PMID- 11860467 TI - Forelimb locomotor generators and quadrupedal locomotion in the neonatal rat. AB - The spinal localization of the forelimb locomotor generators and their interactions with other spinal segments were investigated on in vitro brainstem spinal cord preparations of new-born rats. Superfusion of the cervicothoracic cord (C1-T4) with high K+/low Mg2+ artificial cerebrospinal fluid (aCSF) evoked rhythmic motor root activity that was limited to low cervical (C7, C8) and high thoracic (T1) spinal levels. This activity consisted of synchronous, homolateral bursts and a typical alternating bilateral pattern. Rhythmic activity with similar locomotor-like characteristics could be induced with either serotonin (5 HT, 5 microm), N-methyl-d-aspartate (NMDA, 5 microm), kainate (10 microm) or a "cocktail" of 5-HT (5 microm) and NMDA (5 microm). During 5-HT/NMDA perfusion of the cervicothoracic cord, induced bursting was no longer restricted to C7-T1 levels, but also occurred at cervical C3-C5 levels and with C5-C8 homolateral alternation. Spinal transections between C6 and C7 cervical segments did not abolish rhythmic activity in C7-T1, but suppressed locomotor-like rhythmicity at C3-C5 levels. Reduced regions comprising the C7-C8 or C8-T1 segments maintained rhythmicity. Superfusion of the whole cord with 5-HT/NMDA induced ventral root bursting with similar frequencies at all recorded segments (cervical, thoracic and lumbar). After isolation, the T3-T10 cord was unable to sustain any rhythmic activity while cervical and lumbar segmental levels continued to burst, albeit at different frequencies. We also found that the faster caudal and the slower rostral locomotor generators interact to produce coordinated locomotor-like activity in all segments of the intact spinal cord. In conclusion, C7-T1 spinal levels display a strong motor rhythmogenic ability; with the lumbar generators, they contribute to coordinated rhythmic activity along the entire spinal cord of a quadrupedal locomoting mammal. PMID- 11860468 TI - Hypothalamic-pituitary-adrenal axis function and corticosterone receptor expression in behaviourally characterized young and aged Long-Evans rats. AB - In the current investigation, hypothalamic-pituitary-adrenal (HPA) axis function was examined in young and aged male Long-Evans rats that were initially assessed on a version of the Morris water maze sensitive to cognitive impairment during ageing. In behaviourally characterized rats, a 1-h restraint stress paradigm revealed that plasma corticosterone concentrations in aged cognitively impaired rats took significantly longer to return to baseline following the stressor than did those in young or aged cognitively unimpaired rats. No differences in basal or peak plasma corticosterone concentrations, however, were observed between young or aged rats, irrespective of cognitive status. Using ribonuclease protection assays and in situ hybridization, we evaluated mineralocorticoid receptor (MR) and glucocorticoid receptor (GR) mRNA abundance in young and aged rats characterized on the spatial task. Abundance of MR mRNA was decreased as a function of age in stratum granulosum but not hippocampus proper, and the decrease in MR mRNA was largely unrelated to cognitive status. However, GR mRNA was significantly reduced in several hippocampal subfields (i.e. stratum granulosum and temporal hippocampus proper) and other related cortical structures (medial prefrontal and olfactory regions) of aged cognitively impaired rats compared to either young or aged cognitively unimpaired cohorts, and was significantly correlated with spatial learning ability among the aged rats in each of these brain regions. In agreement with previous stereological data from this ageing model, no changes were detected in neuron density in the hippocampus of the rats used in the in situ hybridization analysis. These data are the first to describe a coordinated decrease in GR mRNA in a functional brain system including hippocampus and related cortical areas that occurs in tandem with impairments of the HPA response to stress and cognitive decline in ageing. PMID- 11860469 TI - Neuroprotective effect of interleukin-6 and IL6/IL6R chimera in the quinolinic acid rat model of Huntington's syndrome. AB - Ciliary neurotrophic factor prevents behavioural deficits and striatal degeneration in rat and primate models of Huntington's disease. Interleukin-6, another member of the cytokine family, and the chimeric molecule (IL6/IL6R) in which interleukin-6 and its soluble receptor are fused, have been shown to exert trophic action on various neuronal populations in the central nervous system. Therefore, we investigated the neuroprotective effect of these two molecules in the quinolinic acid model of Huntington's disease. LacZ-, interleukin-6- and IL6/IL6R-expressing lentiviral vectors were stereotaxically injected into the striatum of Wistar rats. Three weeks later the animals were lesioned through the intrastriatal injection of 180 nmol of quinolinic acid. The extent of the striatal damage was significantly diminished in the rats that had been treated with interleukin-6 or IL6/IL6R. The neuroprotective effect was, however, more pronounced with the IL6/IL6R chimera than with interleukin-6 as indicated by the volume of the lesions (38.6 +/- 10% in the IL6/IL6R group, 63.3 +/- 3.6% in the IL-6 group and 84.3 +/-2.9% in the control group). Quantitative analysis of striatal interneurons further demonstrated that the IL6/IL6R chimera is more neuroprotective than IL-6 on ChAT- and NADPH-d-immunoreactive neurons. These results suggest that the IL6/IL6R chimera is a potential treatment for Huntington's disease. PMID- 11860470 TI - Glial-derived arginine, the nitric oxide precursor, protects neurons from NMDA induced excitotoxicity. AB - Excitotoxic neuronal cell death is characterized by an overactivation of glutamate receptors, in particular of the NMDA subtype, and the stimulation of the neuronal nitric oxide synthase (nNOS), which catalyses the formation of nitric oxide (NO) from l-arginine (L-Arg). At low L-Arg concentrations, nNOS generates NO and superoxide (O2(.)(-)), favouring the production of the toxin peroxynitrite (ONOO-). Here we report that NMDA application for five minutes in the absence of added L-Arg induces neuronal cell death, and that the presence of L-Arg during NMDA application prevents cell loss by blocking O2(.)(-) and ONOO- formation and by inhibiting mitochondrial depolarization. Because L-Arg is transferred from glial cells to neurons upon activation of glial glutamate receptors, we hypothesized that glial cells play an important modulator role in excitotoxicity by releasing L-Arg. Indeed, as we further show, glial-derived L Arg inhibits NMDA-induced toxic radical formation, mitochondrial dysfunction and cell death. Glial cells thus may protect neurons from excitotoxicity by supplying L-Arg. This potential neuroprotective mechanism may lead to an alternative approach for the treatment of neurodegenerative diseases involving excitotoxic processes, such as ischemia. PMID- 11860471 TI - Sciatic nerve regeneration in rats induced by transplantation of in vitro differentiated bone-marrow stromal cells. AB - Bone marrow stromal cells (MSCs) are multipotent stem cells that have the potential to differentiate into bone, cartilage, fat and muscle. We now demonstrate that MSCs can be induced to differentiate into cells with Schwann cell characteristics, capable of eliciting peripheral nervous system regeneration in adult rats. MSCs treated with beta-mercaptoethanol followed by retinoic acid and cultured in the presence of forskolin, basic-FGF, PDGF and heregulin, changed morphologically into cells resembling primary cultured Schwann cells and expressing p75, S-100, GFAP and O4. The MSCs were genetically engineered by transduction with retrovirus encoding green fluorescent protein (GFP), and then differentiated by treatment with factors described above. They were transplanted into the cut ends of sciatic nerves, which then responded with vigorous nerve fibre regeneration within 3 weeks of the operation. Myelination of regenerated fibers by GFP-expressing MSCs was recognized using confocal and immunoelectron microscopy. The results suggest that MSCs are able to differentiate into myelinating cells, capable of supporting nerve fibre re-growth, and they can therefore be applied to induce nerve regeneration. PMID- 11860472 TI - Temporally controlled targeted somatic mutagenesis in the mouse brain. AB - To develop spatio-temporally controlled somatic mutagenesis in the adult mouse nervous system, we established transgenic mice expressing the tamoxifen-inducible Cre-ERT recombinase under the control of the mouse prion protein (PrP) promoter. Cre-ERT was expressed in most regions of the brain and in the retina of one transgenic line, whereas its expression was mostly restricted to the hippocampus and the cerebellum in another line. As tamoxifen efficiently induced Cre-mediated recombination in the various neuronal cell types expressing Cre-ERT in the brain of adult mice, the PrP-Cre-ERT lines should be valuable tools to study the functions of genes involved in neurodegenerative diseases or regeneration, and in complex processes such as behaviour, learning and memory. Some limitations of presently available reporter lines for Cre-mediated recombination in adult mouse CNS are discussed. PMID- 11860473 TI - Changes in P2X receptor responses of sensory neurons from P2X3-deficient mice. AB - Dorsal root ganglion (DRG) neurons respond to ATP with transient, persistent or biphasic inward currents. In contrast, the ATP responses in nodose neurons are persistent. These sustained currents are also heterogeneous, with one component being accounted for by P2X2/3 receptors, and the residual response probably mediated by P2X2 receptors, although the direct evidence for this has been lacking. In the present study, we examined the P2X receptors on DRG and nodose neurons from P2X3-deficient (P2X3-/-) mice, using whole cell voltage-clamp recording and immunohistochemistry. We found that all P2X3-/- DRG neurons lacked rapidly desensitizing response to ATP, and both DRG and nodose neurons from P2X3 null mutant mice no longer responded to alpha,beta-methylene ATP (alphabetameATP). In contrast, ATP evoked persistent inward current in 12% of DRG neurons and 84% of nodose neurons from P2X3-/- mice. This retained persistent response to ATP on nodose neurons had an EC50 for ATP of 77 microm, was antagonized by Cibacron blue and pyridoxal-5-phosphate-6-azophenyl-2',4' disulphonic acid, potentiated by Zn2+ and acidification, but not enhanced by ivermectin or diinosine pentaphosphate. 2',3'-O-Trinitrophenyl-ATP antagonized this response with an IC50 of 8 microm. All these properties are consistent with those of recombinant P2X2 homomeric receptors. Furthermore, specific P2X2 receptor immunoreactivity detected in wild-type sensory neurons was unaltered in null mutant mice. Therefore, the alphabetameATP-insensitive persistent responses on nodose neurons are likely to be mediated by P2X2 homomers, which contribute to 60% of currents evoked by 100 microm ATP in the wild type. PMID- 11860474 TI - HIV-1 protein Tat reduces the glutamate-induced intracellular Ca2+ increase in cultured cortical astrocytes. AB - The trans-activator protein Tat of the human immunodeficiency virus type 1 (HIV 1) is regarded as an injurious molecule in the pathogenesis of HIV-1 associated encephalopathy (HIVE). We investigated the effects of Tat on neuroligand-induced intracellular Ca2+ increase in cultured astroglial cells. Rat cortical astrocytes, human glioblastoma cells and glial restricted precursor cells, from a human embryonic teratocarcinoma cell line, were incubated with recombinant Tat (100 ng/mL for 60 min) which induced a significant reduction of glutamate or ATP induced intracellular Ca2+ increase ("glutamate response", "ATP response"). The reduction of the glutamate response was also observed following cell incubation with cell extracts of HeLa-T4+ cells transiently transfected with an expression plasmid coding for Tat. However, inactivation of the transcriptional trans activity of Tat, by using a mutant form of Tat, as well as inhibition of de novo protein synthesis by cycloheximide abolished the effect on the glutamate response. This suggests that Tat acts upon induction of a so far unknown cellular gene whose gene product causes the reduction of glutamate responses. As the effect of Tat resembles the effect of TNFalpha on glutamate responses [Koller et al. (2001) Brain Res., 893, 237-243] which is locally released within the brains of HIVE patients, we also tested for synergistic effects of Tat and TNFalpha on the glutamate response. Low concentrations of Tat in combination with subthreshold concentrations of TNFalpha also elicited a marked reduction of astroglial glutamate responses. Our data suggest that Tat and TNFalpha, both by itself and synergistically, induce astroglial dysfunction. PMID- 11860475 TI - Acute modulation of synaptic transmission to motoneurons by BDNF in the neonatal rat spinal cord. AB - We investigated the acute effects of bath applied BDNF on synaptic input to motoneurons in the hemisected spinal cord of the neonatal rat. Motoneurons were recorded intracellularly, and BDNF-induced modulation of the synaptic response to stimulation of the homologous dorsal root (DR) and the ventrolateral funiculus (VLF) was examined. All motoneurons exhibited long-lasting (up to several hours) depression of the DR-activated monosynaptic AMPA/kainate-receptor mediated EPSP in response to BDNF but in about half of the motoneurons this was preceded by facilitation. VLF-evoked AMPA/kainate EPSPs in the same motoneurons were unaffected. BDNF effects were blocked by K252a and were not observed in neonates older than 1 week. Bath applied NMDA antagonists APV and MK-801 abolished both facilitatory and inhibitory actions of BDNF on the AMPA/kainate responses indicating the requirement for functional NMDA receptors. The pharmacologically isolated, DR-evoked, NMDA receptor-mediated response exhibited the same pattern of changes after BDNF superfusion. When introduced into the motoneuron through the recording microelectrode, MK-801 selectively blocked the facilitatory action of BDNF. Furthermore, BDNF enhanced NMDA-induced depolarization of the motoneuron in the presence of tetrodotoxin (TTX), thus, confirming its facilitatory effect on motoneuron NMDA receptors. Bath application of either BDNF or NMDA depressed the monosynaptic EPSP after selective blockade of postsynaptic NMDA receptors indicating a role for presynaptic NMDA receptors in BDNF-induced inhibitory action. Thus, BDNF-induced facilitation of monosynaptic EPSPs in neonatal rats is mediated by direct effects on postsynaptic NMDA receptors, while its inhibitory action occurs presynaptically. PMID- 11860476 TI - Evidence that interleukin-1beta and reactive oxygen species production play a pivotal role in stress-induced impairment of LTP in the rat dentate gyrus. AB - Long-term potentiation (LTP) in both area CA1 and the dentate gyrus is attenuated by stress and the evidence is consistent with the view that this is a consequence of increased activation of glucocorticoid receptors, in the hippocampus, following the stress-induced increase in circulating corticosterone. It has been shown that expression of the pro-inflammatory cytokine, interleukin-1beta (IL 1beta), is increased in hippocampus in response to stress; this finding together with the observation that IL-1beta exerts an inhibitory effect on LTP, suggests that IL-1beta may play a key role in mediating this inhibitory effect of stress on LTP. In this study, we explore this possibility and report that stress is also associated with increased reactive oxygen species production. The evidence presented supports the view that this is secondary to the stress-induced increase in IL-1beta concentration, as IL-1beta increased activity of superoxide dismutase and increased reactive oxygen species accumulation in hippocampus in vitro. We report that the inhibitory effect of stress on LTP is mimicked by H2O2, which increases reactive oxygen species accumulation, and by IL-1beta, the effect of which is overcome by the antioxidant, phenylarsine oxide. The hypothesis that the stress-induced increase in reactive oxygen species production may underlie the suppression of LTP is further supported by the finding that the effect of stress is abrogated by dietary manipulation with antioxidant vitamins E and C. PMID- 11860478 TI - Increased cannabinoid CB1 receptor binding and activation of GTP-binding proteins in the basal ganglia of patients with Parkinson's syndrome and of MPTP-treated marmosets. AB - Recent evidence obtained in rat models of Parkinson's disease showed that the density of cannabinoid CB1 receptors and their endogenous ligands increase in basal ganglia. However, no data exists from post-mortem brain of humans affected by Parkinson's disease or from primate models of the disorder. In the present study, we examined CB1 receptor binding and the magnitude of the stimulation by WIN55,212-2, a specific CB1 receptor agonist, of [35S]GTPgammaS binding to membrane fractions from the basal ganglia of patients affected by Parkinson's disease. In Parkinson's disease, WIN55,212-2-stimulated [35S]GTPgammaS binding in the caudate nucleus, putamen, lateral globus pallidus and substantia nigra was increased, thus indicating a more effective activation of GTP-binding protein coupled signalling mechanisms via CB1 receptors. This was accompanied by an increase in CB1 receptor binding in the caudate nucleus and the putamen, although no changes were observed in the lateral globus pallidus and the substantia nigra. Because Parkinson's disease patients had been chronically treated with l-DOPA, brains were studied from normal common marmosets and 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP)-treated animals with and without chronic L-DOPA treatment. MPTP-lesioned marmosets had increased CB1 receptor binding in the caudate nucleus and the putamen compared to control marmosets, as well as increased stimulation of [35S]GTPgammaS binding by WIN55,212-2. However, following l-DOPA treatment these parameters returned towards control values. The results indicate that a nigro-striatal lesion is associated with an increase in CB1 receptors in the basal ganglia in humans and nonhuman primates and that this increase could be reversed by chronic l-DOPA therapy. The data suggest that CB1 receptor blockade might be useful as an adjuvant for the treatment of parkinsonian motor symptoms. PMID- 11860477 TI - The ubiquitin-proteasome cascade is required for mammalian long-term memory formation. AB - It has been recently demonstrated that ubiquitin-proteasome-mediated proteolysis is required for long-term synaptic facilitation in Aplysia. Here we show that the hippocampal blockade of this proteolytic pathway is also required for the formation of long-term memory in the rat. Bilateral infusion of lactacystin, a specific proteasome inhibitor, to the CA1 region caused full retrograde amnesia for a one-trial inhibitory avoidance learning when given 1, 4 or 7h, but not 10 h, after training. Proteasome inhibitor I produced similar effects. In addition, inhibitory avoidance training resulted in an increased ubiquitination and 26S proteasome proteolytic activity and a decrease in the levels of IkappaB, a substrate of the ubiquitin-proteasome cascade, in hippocampus 4 h after training. Together, these findings indicate that the ubiquitin-proteasome cascade is crucial for the establishment of LTM in the behaving animal. PMID- 11860479 TI - Unilateral lesion of the nigrostriatal pathway induces an increase of neuronal activity of the pedunculopontine nucleus, which is reversed by the lesion of the subthalamic nucleus in the rat. AB - The role of the pedunculopontine nucleus (PPN) in the pathophysiology of Parkinson's disease is still unclear. Using microrecordings, we investigated the changes occurring in PPN neurons after lesions of the substantia nigra compacta (SNc) and the role of the subthalamic nucleus (STN) in these changes. In normal rats the firing rate of PPN neurons was 10.6 +/- 1.4 spikes/s, the majority of neurons (91%) having a regular firing pattern, 6% irregular and 3% in bursts. In rats with 6-hydroxydopamine lesions of the SNc, the firing rate increased significantly to 18.3 +/- 3.0 spikes/s compared with normal rats. In addition, the firing pattern changed significantly: 70% of the neurons discharged regularly, 27% irregularly and 3% in bursts. In rats with ibotenic acid lesions of the STN, the firing rate decreased significantly to 7.2 +/- 0.9 spikes/s and the firing pattern changed significantly: 50% of the neurons discharged regularly, 43% irregularly and 7% in bursts. The rats with combined SNc and STN lesions showed no change in the firing rate (8.5 +/- 1.0 spikes/s) compared to normal rats. The firing pattern changed significantly: 69% of the cells discharged regularly, 26% irregularly and 5% in bursts. These findings demonstrate that PPN neurons are overactive and more irregular in the 6 hydroxydopamine-lesioned rats, suggesting the implication of this nucleus in the pathophysiology of parkinsonism. Moreover, the fact that STN lesions induced a reduction in the firing rate of the PPN in normal rats and a normalization of the firing rate in rats with 6-hydroxydopamine lesions suggests that this nucleus is under major control of the STN. PMID- 11860480 TI - Alterations in behaviour and glutamate transmission following presentation of stimuli previously associated with cocaine exposure. AB - To study the role of glutamate in cocaine-conditioned responses, we developed a rat model in which conditioned locomotion is produced by repeated pairing of cocaine with discrete stimuli (flashing light and metronome). "Paired" subjects received cocaine (15 mg/kg) prior to six exposures to stimuli for 30 min in the test environment. "Unpaired" subjects received equivalent presentations of the stimuli yet received cocaine in home cages. Tests with the stimuli alone demonstrated that the conditioned locomotion displayed by Paired subjects was evident at 3 or 10 days post-training and resistant to two sessions of testing. The degree of conditioned locomotion was not correlated with the subjects' response to novelty or cocaine. Administration of the noncompetitive AMPA receptor antagonist GYKI 52466 (2.5 mg/kg, a dose without effect on spontaneous activity) attenuated the expression of conditioned activity. In vivo microdialysis revealed that Paired subjects had significantly lower basal glutamate levels in the nucleus accumbens (NAc) than did Unpaired subjects when no stimuli were presented. Presentation of the conditioned stimuli resulted in significant increases in glutamate levels in the NAc in the Paired group whilst glutamate levels in the Unpaired group remained unchanged. The associative control of glutamate levels in the NAc by stimuli formerly paired with a drug of abuse is an unprecedented finding. It is likely to reflect the convergence of excitatory inputs that the NAc receives from limbic structures. PMID- 11860481 TI - Cholinergic and noncholinergic septal neurons modulate strategy selection in spatial learning. AB - Rats solving a simple spatial discrimination task in a plus maze initially employ a place-learning strategy, then switch to a motor response strategy. The hippocampus is required for the use of a place-learning strategy in this task. Rats with 192 IgG-saporin lesions of the medial septum/vertical limb of the diagonal band (MS/VDB), that selectively removed cholinergic neurons projecting to the hippocampus, were significantly facilitated in acquisition of the spatial discrimination, and switched from place to response strategies just as control rats did. Rats with ibotenic acid lesions of the MS/VDB, that produced cell loss in the MS/VDB but little damage to cholinergic neurons, were significantly impaired in acquiring the spatial discrimination and did not reliably employ either a place or response strategy at any point in training. This suggests that the MS/VDB modulates hippocampal involvement in place learning, but that cholinergic MS/VDB neurons are neither necessary nor sufficient for using a place strategy to solve a spatial discrimination. PMID- 11860482 TI - Diversity of receptive field changes in auditory cortex during natural sleep. AB - Twenty years ago, the study by Livingstone and Hubel [(1981) Nature, 291, 554] was viewed as a first step toward understanding how changes in state of vigilance affect sensory processing. Since then, however, very few attempts have been made to progress in this direction. In the present study, 56 cells were recorded in the auditory cortex of adult, undrugged guinea pigs, and the frequency tuning curves were tested during continuous and stable periods of wakefulness and of slow-wave sleep (SWS). Twelve cells were also tested during paradoxical sleep. Over the whole cell population, the response latency, the frequency selectivity and the size of the suprathreshold receptive field were not significantly modified during SWS compared with waking. However, this lack of global effects resulted from the heterogeneity of response changes displayed by cortical cells. During SWS, the receptive field size varied as a function of the changes in evoked responses: it was unchanged for the cells whose evoked responses were not modified (38% of the cells), reduced for the cells whose responses were decreased (48%) and enlarged for the cells whose responses were increased (14%). This profile of changes differs from the prevalent receptive field shrinkage that was observed in the auditory thalamus during SWS [Edeline et al. (2000), J. Neurophysiol., 84, 934]. It also contrasts with the receptive field enlargement that was described under anaesthesia when the EEG spontaneously shifted from a desynchronized to a synchronized pattern [Worgotter et al. (1998), Nature, 396, 165]. Reasons for these differences are discussed. PMID- 11860483 TI - Neural interactions between motor cortical hemispheres during bimanual and unimanual arm movements. AB - Cortico-cortical connections through the corpus callosum are a major candidate for mediating bimanual coordination. However, aside from the deficits observed after lesioning this connection, little positive evidence indicates its function in bimanual tasks. In order to address this issue, we simultaneously recorded neuronal activity at multiple sites within the arm area of motor cortex in both hemispheres of awake primates performing different bimanual and unimanual movements. By employing an adapted form of the joint peri-stimulus time histogram technique, we discovered rapid movement-related correlation changes between the local field potentials (LFPs) of the two hemispheres that escaped detection by time-averaged cross-correlation methods. The frequency and amplitude of dynamic modifications in correlations between the hemispheres were similar to those within the same hemisphere. As in previous EEG studies, we found that, on average, correlation decreased during movements. However, a subset of recording site pairs did show transiently increased correlations around movement onset (57% of all pairs and conditions in monkey G, 39% in monkey P). In interhemispheric pairs, these increases were consistently related to the mode of coupling between the two arms. Both the correlations between the movements themselves and the interhemispheric LFP correlation increases were strongest during bimanual symmetric movements, and weaker during bimanual asymmetric and unimanual movements. Increased correlations occurred mainly around movement onset, whilst decreases in correlation dominated during movement execution. The task-specific way in which interhemispheric correlations are modulated is compatible with the notion that interactions between the hemispheres contribute to behavioural coupling between the arms. PMID- 11860484 TI - Role of the corpus callosum in bimanual coordination: a comparison of patients with congenital and acquired callosal damage. AB - The objective of the study was to investigate temporal control in patients with congenital as compared to acquired pathology of the corpus callosum during two different bimanual paradigms: (i) a drawer-opening task during which one hand opened a drawer while the other hand reached and grasped a small object, and (ii) rhythmical circling movements that were executed according to the in-phase or antiphase mode. Synchronization values revealed that patients with acquired callosal dysfunction generally showed optimal behaviour during the goal-directed and familiar drawer-opening task but demonstrated strong tendencies towards desynchronization during circling movements, which became most apparent for antiphase coordination. Whereas one patient with callosal agenesis showed a similar performance, the other acallosal patients performed both activities successfully. These observations indicate that patients with congenital absence of the corpus callosum can make use of compensatory mechanisms for allowing temporal synchronization during bimanual movements whereas patients with acquired callosal dysfunction are severely hampered when the task places significant demands on the control processes. The data also underline that the ability of callosal patients to precisely time events in coordinated actions depend on the task constraints. PMID- 11860485 TI - Neuronal coordination of arm and leg movements during human locomotion. AB - We aimed to study the neuronal coordination of lower and upper limb muscles. We therefore evaluated the effect of small leg displacements during gait on leg and arm muscle electromyographic (EMG) activity in walking humans. During walking on a split-belt treadmill (velocity 3.5 km/h), short accelerations or decelerations were randomly applied to the right belt during the mid or end stance phase. Alternatively, trains of electrical stimuli were delivered to the right distal tibial nerve. The EMG activity of the tibialis anterior (TA), gastrocnemius medialis (GM), deltoideus (Delt), triceps (Tric) and biceps brachii (Bic) of both sides was analysed. For comparison, impulses were also applied during standing and sitting. The displacements were followed by specific patterns of right leg and bilateral arm muscle EMG responses. Most arm muscle responses appeared with a short latency (65-80 ms) and were larger in Delt and Tric than in Bic. They were strongest when deceleration impulses were released during mid stance, associated with a right compensatory TA response. A similar response pattern in arm muscles was obtained following tibial nerve stimulation. The arm muscle responses were small or absent when stimuli were applied during standing or sitting. The arm muscle responses correlated more closely with the compensatory TA than with the compensatory GM responses. The amplitude of the responses in most arm muscles correlated closely with the background EMG activity of the respective arm muscle. The observations suggest the existence of a task-dependent, flexible neuronal coupling between lower and upper limb muscles. The stronger impact of leg flexors in this interlimb coordination indicates that the neuronal control of leg flexor and extensor muscles is differentially interconnected during locomotion. The results are compatible with the assumption that the proximal arm muscle responses are associated with the swinging of the arms during gait, as a residual function of quadrupedal locomotion. PMID- 11860486 TI - Associative effects of Pavlovian differential inhibition of behaviour. AB - The associative inhibitory control of behaviour is a major component of Pavlovian learning theory, but little is known about its functional neuroanatomy. The associative effects of differential inhibition of conditioned behaviour were investigated by mapping learning-related changes in brain activity of the rat with fluorodeoxyglucose autoradiography. Of interest was how a tone is processed in auditory and extra-auditory systems of the rat brain under similar behavioural, but different associative conditions. Conditioned emotional suppression to drink was used to assess training, and summation tests were used to verify that the tone became an inhibitor of conditioned behaviour. In the Inhibitor group, presentations of a tone stimulus alone were intermixed with presentations of a light stimulus followed by footshock. In the Pseudorandom group, the same numbers of tone, light and footshock presentations were used, but they were presented in a pseudorandom fashion. After training, fluorodeoxyglucose uptake was measured during tone presentations. Behavioural responding to the tone was similar during fluorodeoxyglucose uptake in the two groups, yet associative effects were found in brain activity. In the auditory system, the tone produced reduced fluorodeoxyglucose uptake in major relay nuclei (cochlear nucleus and inferior colliculus) in the Inhibitor group relative to the Pseudorandom group. The tone inhibitor produced similar decreases in the septohippocampal system and the retrosplenial cortex. In contrast, the tone inhibitor produced activity increases in somatosensory and reticulocerebellar systems. The findings provide the first detailed map of neural regions involved in the learned associations controlling differential inhibition of conditioned behaviour. PMID- 11860487 TI - Expression of ephrin-A2 in the superior colliculus and EphA5 in the retina following optic nerve section in adult rat. AB - The vertebrate retina projects topographically to visual brain centres. In the developing visual system, gradients of ephrins and Eph receptors play a role in defining topography. At maturity, ephrins but not Ephs are downregulated. Here we show that optic nerve section in adult rat differentially regulates the expression of ephrin-A2 in the superior colliculus (SC) and of EphA5 in the retina. Expression was quantified immunohistochemically; ephrin-A2 levels were also estimated by semiquantitative reverse transcriptase polymerase chain reaction. In the normal SC, ephrin-A2 was expressed at low levels. At 1 month, levels of protein and of mRNA were upregulated across the contralateral SC giving rise to an increasing rostro-caudal gradient. At 6 months, levels had fallen but a gradient remained. In the retina of normal animals, EphA5 was expressed as an increasing naso-temporal gradient. By 1 month, expression was decreased in far temporal retina, resulting in a uniform expression across the naso-temporal axis. We suggest that denervation-induced plastic changes within the SC modify expression of these molecules. PMID- 11860488 TI - Activity- and age-dependent GABAergic synaptic plasticity in the developing rat hippocampus. AB - Activity-dependent plasticity of GABAergic synaptic transmission was investigated in rat hippocampal slices obtained between postnatal day (P) 0-15 using the whole cell patch-clamp recording technique. Spontaneous GABA(A) receptor-mediated postsynaptic currents (sGABA(A)-PSCs) were isolated in the presence of ionotropic glutamate receptor antagonists. A conditioning protocol relevant to the physiological condition, consisting of repetitive depolarizing pulses (DPs) at 0.1 Hz, was able to induce long-lasting changes in both frequency and amplitude of sGABA(A)-PSCs between P0 and P8. Starting from P12, DPs were unable to induce any form of synaptic plasticity. The effects of DPs were tightly keyed to the frequency at which they were delivered. When delivered at a lower (0.05 Hz) or higher (1 Hz) frequency, DPs failed to induce any long-lasting change in the frequency or amplitude of sGABA(A)-PSCs. In two cases, DPs were able to activate sGABA(A)-PSCs in previously synaptically silent cells at P0-1. These results show that long-term changes in GABAergic synaptic activity can be induced during a restricted period of development by a conditioning protocol relevant to the physiological condition. It is suggested that such activity-induced modifications may represent a physiological mechanism for the functional maturation of GABAergic synaptic transmission. PMID- 11860489 TI - The voltage-sensitive motor protein and the Ca2+-sensitive cytoskeleton in developing rat cochlear outer hair cells. AB - Cochlear outer hair cells (OHCs) possess a unique fast voltage-driven motility associated with a voltage-sensitive motor protein embedded in the basolateral membrane. This mechanism is believed to underlie the cochlear amplification in mammals. OHCs also have a Ca2+/calmodulin-dependent mechanical pathway which involves a submembranous circumferential cytoskeleton. The purpose of this study was to compare the functional appearance of the voltage-sensitive motor proteins with that involving the Ca2+-sensitive cytoskeleton during postnatal development of rat OHCs. We demonstrate that whole-cell electromotility and Ca2+-voked mechanical responses, by ionomycin, develop concomitantly after postnatal day 5 (P5). These two mechanical properties also develop simultaneously in OHCs isolated from two-week-old cultures of P0-P1 organs of Corti. This excludes the participation of neural innervation in the postnatal maturation of the OHCs' motile properties. In addition, we show that the expression of the membranous voltage-sensitive motor protein precedes, by several days, the appearance of whole-cell electromotility. The concomitant development of whole-cell electromotility and Ca2+-sensitive motility, both in vivo and in vitro, underlines the cytoskeleton as an important factor in the functional organization of the voltage-sensitive motor proteins within the plasma membrane. PMID- 11860490 TI - AMPA receptor activation induces association of G-beta protein with the alpha subunit of the sodium channel in neurons. AB - Glutamatergic transmission is mediated by ionotropic receptors that directly gate cationic channels and metabotropic receptors that are coupled to second messenger generating systems and to ionic channels via heterotrimeric guanine-nucleotide binding- (G) proteins. This distinction cannot be made for the ionotropic receptor subclass activated by alpha-amino-3-hydroxy-5-methylisoxazole-4 propionic acid (AMPA), which has been shown to be physically associated with the alpha-subunit of Gi1 protein and activates this G-protein. Here, we report that, in addition to a Ca2+ influx, AMPA induces the mobilization of Ca2+ from the mitochondrial pool by reversing the mitochondrial Na+/Ca2+ exchanger in mouse neurons in primary culture. Both processes required the activation of tetrodotoxin-sensitive Na+ channels. AMPA receptor activation modified the gating properties of the Na+ channel, independently of the AMPA current, suggesting a G protein-mediated process. Indeed, co-immunoprecipitation experiments indicated that AMPA receptor activation induced the association of Gbeta with the alpha subunit of the Na+ channel. These results suggest that, in addition to its ionic channel function, the AMPA receptor is coupled to Na+ channels through G-proteins and that this novel metabotropic function is involved in the control of neuronal excitability. PMID- 11860491 TI - Anti-inflammatory antioxidants attenuate the expression of inducible nitric oxide synthase mediated by advanced glycation endproducts in murine microglia. AB - Advanced glycation endproducts (AGEs) accumulate on long-lived protein deposits including beta-amyloid plaques in Alzheimer's disease (AD). AGE-modified amyloid deposits contain oxidized and nitrated proteins as markers of a chronic neuroinflammatory condition and are surrounded by activated microglial and astroglial cells. We show in this study that AGEs increase nitric oxide production by induction of the inducible nitric oxide synthase (iNOS) on the mRNA and protein level in the murine microglial cell line N-11. Membrane permeable antioxidants including oestrogen derivatives (e.g. 17beta-oestradiol) thiol antioxidants (e.g. (R+)-alpha-lipoic acid) and Gingko biloba extract EGb 761, but not phosphodiesterase inhibitors such as propentophylline, prevent the up regulation of AGE-induced iNOS expression and NO production. These results indicate that oxygen free radicals serve as second messengers in AGE-induced pro inflammatory signal transduction pathways. As this pharmacological mechanism is not only relevant for Alzheimer's disease, but also for many chronic inflammatory conditions, such membrane-permeable antioxidants could be regarded not only as antioxidant, but also as potent therapeutic anti-inflammatory drugs. PMID- 11860492 TI - Protracted expression of serotonin transporter and altered thalamocortical projections in the barrelfield of hypothyroid rats. AB - In humans, thyroid hormone deficiency during development causes severe neurological diseases but the underlying mechanisms are unclear. We have examined the effects of thyroid hormones on the development of somatosensory thalamocortical projections, by inducing hypothyroidism in rats by methimazole treatment at embryonic day 13 and subsequent thyroidectomy at postnatal day 6 (P6). Initial development of the thalamocortical projections and their tangential and laminar patterning were similar in normal and hypothyroid rats from birth to P4. The tangential spread of the thalamocortical arbors is reduced in hypothyroid rats after P4, paralleling the overall cortical atrophy. Anterograde tracing and single axon reconstructions indicate that thalamic afferents reached layer IV but that they had fewer and shorter branches, with a 42% reduction in the number of boutons. The transient serotonin (5-HT) immunostaining and 5-HT transporter (5 HTT) expression were both prolonged by 5 days in hypothyroid rats. This does not reflect a delayed maturation of the thalamus because other transiently expressed genes such as the vesicular monoamine transporter and the 5-HT1B receptor are not modified. Protracted 5-HTT expression also occurred in other areas with transient expression, but no changes were observed in the raphe nuclei where the 5-HTT is expressed permanently. Thus, thyroid hormones appear to be important in regulating the extinction of the 5-HTT in nonserotoninergic neurons. The transient stabilization of 5-HT reuptake in hypothyroid rats could affect the growth of thalamic axons. Our data stress the importance of maternal and foetal thyroid hormones for the normal development of sensory systems. PMID- 11860493 TI - Selective cardiorespiratory and catecholaminergic areas express the hypoxia inducible factor-1alpha (HIF-1alpha) under in vivo hypoxia in rat brainstem. AB - Under severe oxygen deprivation, all cells are able to express the transcription factor HIF-1, which activates a wide range of genes. Under tolerable hypoxia, chemosensory inputs are integrated in brainstem areas, which control cardiorespiratory responses. However, the molecular mechanisms of this functional acclimatization are unknown. We investigated when and where the inducible HIF 1alpha subunit is expressed in the rat brainstem in vivo, under physiological hypoxia. The regional localization of HIF-1alpha mRNA and protein was determined by in situ hybridization and immunocytochemistry in adult male rats exposed to moderate hypoxia (10% O2) for 1-6 h. HIF-1alpha protein was found in cell types identified by immunocytochemistry as catecholaminergic neurons. Hypoxia induced HIF-1alpha mRNA and protein in only some parts of the brainstem located dorsomedially and ventrolaterally, which are those involved in the cardiorespiratory control. No labelling was detected under normoxia. The protein was detected in glia and neurons after 1 and 6 h of hypoxia, respectively. A subset of A2C2 and A1C1 catecholaminergic neurons colocalized tyrosine hydroxylase and HIF-1alpha proteins under hypoxia, but no HIF-1alpha was detected in more rostral catecholaminergic areas. In contrast to cardiorespiratory areas, HIF-1alpha protein was already present under normoxia in glial cells of brainstem tracts but was not overexpressed under hypoxia, although HIF-1alpha mRNA was up regulated. In conclusion, there appear to be two regulatory mechanisms for HIF 1alpha expression in the brainstem: hypoxic induction of HIF-1alpha protein in cardiorespiratory-related areas and constitutive protein expression unaffected by hypoxia in brainstem tracts. PMID- 11860494 TI - Estrogen and exercise interact to regulate brain-derived neurotrophic factor mRNA and protein expression in the hippocampus. AB - We investigated the possibility that estrogen and exercise interact in the hippocampus and regulate brain-derived neurotrophic factor (BDNF), a molecule increasingly recognized for its role in plasticity and neuron function. An important aspect of this study is to examine the effect of different time intervals between estrogen loss and estrogen replacement intervention. We demonstrate that in the intact female rat, physical activity increases hippocampal BDNF mRNA and protein levels. However, the exercise effect on BDNF up regulation is reduced in the absence of estrogen, in a time-dependent manner. In addition, voluntary activity itself is stimulated by the presence of estrogen. In exercising animals, estrogen deprivation reduced voluntary activity levels, while estrogen replacement restored activity to normal levels. In sedentary animals, estrogen deprivation (ovariectomy) decreased baseline BDNF mRNA and protein, which were restored by estrogen replacement. Despite reduced activity levels in the ovariectomized condition, exercise increased BDNF mRNA levels in the hippocampus after short-term (3 weeks) estrogen deprivation. However, long-term estrogen-deprivation blunted the exercise effect. After 7 weeks of estrogen deprivation, exercise alone no longer affected either BDNF mRNA or protein levels. However, exercise in combination with long-term estrogen replacement increased BDNF protein above the effects of estrogen replacement alone. Interestingly, protein levels across all conditions correlated most closely with mRNA levels in the dentate gyrus, suggesting that expression of mRNA in this hippocampal region may be the major contributor to the hippocampal BDNF protein pool. The interaction of estrogen, physical activity and hippocampal BDNF is likely to be an important issue for maintenance of brain health, plasticity and general well-being, particularly in women. PMID- 11860495 TI - Steroid structure and pharmacological properties determine the anti-amnesic effects of pregnenolone sulphate in the passive avoidance task in rats. AB - Pregnenolone sulphate (PREGS) has generated interest as one of the most potent memory-enhancing neurosteroids to be examined in rodent learning studies, with particular importance in the ageing process. The mechanism by which this endogenous steroid enhances memory formation is hypothesized to involve actions on glutamatergic and GABAergic systems. This hypothesis stems from findings that PREGS is a potent positive modulator of N-methyl-d-aspartate receptors (NMDARs) and a negative modulator of gamma-aminobutyric acid(A) receptors (GABA(A)Rs). Moreover, PREGS is able to reverse the amnesic-like effects of NMDAR and GABA(A)R ligands. To investigate this hypothesis, the present study in rats examined the memory-altering abilities of structural analogs of PREGS, which differ in their modulation of NMDAR and/or GABA(A)R function. The analogs tested were: 11 ketopregnenolone sulphate (an agent that is inactive at GABA(A)Rs and NMDARs), epipregnanolone ([3beta-hydroxy-5beta-pregnan-20-one] sulphate, an inhibitor of both GABA(A)Rs and NMDARs), and a newly synthesized (-) PREGS enantiomer (which is identical to PREGS in effects on GABA(A)Rs and NMDARs). The memory-enhancing effects of PREGS and its analogs were tested in the passive avoidance task using the model of scopolamine-induced amnesia. Both PREGS and its (-) enantiomer blocked the effects of scopolamine. The results show that, unlike PREGS, 11 ketopregnenolone sulphate and epipregnanolone sulphate failed to block the effect of scopolamine, suggesting that altering the modulation of NMDA receptors diminishes the memory-enhancing effects of PREGS. Moreover, enantioselectivity was demonstrated by the ability of natural PREGS to be an order of magnitude more effective than its synthetic enantiomer in reversing scopolamine-induced amnesia. These results identify a novel neuropharmacological site for the modulation of memory processes by neuroactive steroids. PMID- 11860496 TI - Cerebellar contribution to spatial event processing: involvement in procedural and working memory components. AB - Spatial function is one of the cognitive functions altered in the presence of cerebellar lesions. We investigated the cerebellar contribution to the acquisition of spatial procedural and working memory components by means of a radial maze. To establish whether a cerebellar lesion would cause a deficit in solving the radial maze, a first experiment was carried out by using a full baited maze procedure in different experimental groups, with or without cerebellar lesion and with or without pretraining. Non-pretrained hemicerebellectomized (HCbed) animals exhibited impaired performances in all (motor, spatial and procedural) task aspects. Pre-trained HCbed animals performed similarly to control animals in the task aspects linked to the processing of spatial and procedural factors. To distinguish procedural from working memory components, a forced-choice paradigm of the radial maze was used in the second experiment. Non-pretrained HCbed rats continued to make a lot of errors and show severe perseverative tendencies, already observed in the first experiment, supporting a specific cerebellar role in acquiring new behaviours and in modifying them in relation to the context. Interestingly, hindered from putting the acquired explorative patterns into action and compelled to use only working memory abilities, the pretrained HCbed group exhibited a dramatic worsening of performance. In conclusion, the present findings demonstrate that cerebellar damage induces a specific behaviour in radial maze tasks, characterized by an inflexible use of the procedures (if indeed any procedure was acquired before the lesion) and by a severe impairment in working memory processes. PMID- 11860497 TI - Impaired fear conditioning but enhanced seizure sensitivity in rats given repeated experience of withdrawal from alcohol. AB - Repeated experience of withdrawal from chronic alcohol treatment increases sensitivity to seizures. It has been argued by analogy that negative affective consequences of withdrawal also sensitize, but repeated experience of withdrawal from another sedative-hypnotic drug, diazepam, results in amelioration of withdrawal anxiety and aversiveness. We tested whether giving rats repeated experience of withdrawal from alcohol altered their ability to acquire a conditioned emotional response (CER). Male Hooded Lister rats were fed a nutritionally complete liquid diet as their only food source. Different groups received control diet, or diet containing 7% ethanol. Rats receiving ethanol diet were fed for either 24 days (Single withdrawal, SWD), or 30 days, with two periods of 3 days, starting at day 11, and 21, in which they received control diet (Repeated withdrawal, RWD). All rats were fed lab chow at the end of their liquid diet feeding period. Starting 12 days after the final withdrawal, groups of Control, SWD and RWD rats were given pentylenetetrazole (PTZ; 30 mg/kg, i.p.) three times a week, and scored for seizures. The occurrence of two successive Stage 5 seizures was taken as the criterion for full PTZ kindling. Other groups of control, SWD and RWD rats were trained to operate levers to obtain food, and were then exposed, in a fully counterbalanced design, to light and tone stimuli which predicted unavoidable footshock (CS+), or which had no consequences (CS-). Rats consumed approximately 17.5 g/kg/day of ethanol, resulting in blood alcohol levels of approximately 100 mg/dL. Repeated administration of PTZ resulted in increasing seizure scores. RWD rats achieved kindling criterion faster than either Control or SWD rats. No differences were seen in the groups in flinch threshold to footshock (0.3 mA). At a shock intensity of 0.35 mA, Control, but not RWD or SWD rats showed significant suppression to the CS+ CS- presentation did not affect response rates. The three groups differed in their response to pairing the CS+ with increasing shock levels, the Controls remaining more sensitive to the CS+. SWD rats showed significant suppression of lever pressing during CS+ presentations only at 0.45 and 0.5 mA, and RWD rats only at 0.5 mA. Giving rats repeated experience of withdrawal from chronic ethanol results in increased sensitivity to PTZ kindling, but reduces their ability to acquire a CER. Withdrawal kindling of sensitivity to anxiogenic events does not seem to occur under circumstances which give rise to kindling of seizure sensitivity. PMID- 11860498 TI - Stabilization of mutant Cu/Zn superoxide dismutase (SOD1) protein by coexpressed wild SOD1 protein accelerates the disease progression in familial amyotrophic lateral sclerosis mice. AB - Transgenic mice carrying familial amyotrophic lateral sclerosis (FALS)-linked mutant Cu/Zn superoxide dismutase (SOD1) genes such as G93A (G93A-mice) and G85R (G85R-mice) genes develop limb paresis. Introduction of human wild type SOD1 (hWT SOD1) gene, which does not cause motor impairment by itself, into different FALS mice resulted in different effects on their clinical courses, from no effect in G85R-mice to acceleration of disease progression in G93A-mice. However, the molecular mechanism which causes the observed difference, has not been clarified. We hypothesized that the difference might be caused by the stability of mutant SOD1 proteins. Using a combination of mass spectrometry and enzyme-linked immunosorbent assay, we found that the concentration of G93A-SOD1 protein was markedly elevated in tissues of transgenic mice carrying both G93A- and hWT-SOD1 genes (G93A/hWT-mice) compared to that in G93A-mice, and also found that the concentration of G93A-SOD1 protein had a close relation to the disease duration. The concentration of metallothionein-I/II in the spinal cord, reflecting the degree of copper-mediated oxidative stress, was highest in G93A/hWT-mice, second in G93A-mice, and normal in the mice carrying hWT-SOD1 gene. These results indicated that the increase of G93A-SOD1 protein was responsible for the increase of oxidative stress and disease acceleration in G93A/hWT-mice. We speculate that coexpression of hWT-SOD1 protein is deleterious to transgenic mice carrying a stable mutant such as G93A-SOD1, because this mutant protein is stabilized by hWT SOD1 protein, but not to transgenic mice carrying an unstable mutant such as G85R SOD1, because this mutant protein is not stabilized by hWT-SOD1. PMID- 11860499 TI - Seizure-induced gene expression in area CA1 of the mouse hippocampus. AB - Synaptic plasticity in the hippocampus requires activity-dependent gene expression. We have therefore profiled gene expression in area CA1 following the induction of an electroshock-evoked maximal seizure. Using cDNA microarrays, the differential expression of approximately 9000 cDNAs was examined. In situ hybridization on 14 transcripts that showed strongest modulation in the microarray screen (1.8-2-fold) confirmed the differential expression of a single gene that encodes for the nuclear hormone receptor NGFI-B (Nur77, N10). Although this gene is only modestly up-regulated (approximately 2-fold) in area CA1, in situ hybridization revealed that maximal seizures induce a marked (approximately 12-fold) up-regulation of NGFI-B in the dentate gyrus. These data support the notion [French et al. (2001) Eur. J. Neurosci., 13, 968-976] that CA1 pyramidal neurons are more refractory than granule cells of the dentate gyrus with respect to activity-dependent gene transcription. Furthermore, our results argue against a large cohort of activity-dependent genes in area CA1. PMID- 11860501 TI - Hippocampal granule neuron production and population size are regulated by levels of bFGF. AB - Numerous studies of the proliferative effects of basic fibroblast growth factor (bFGF) in culture, including neonatal and adult hippocampal precursors, suggest that the factor plays a ubiquitous and life-long role in neurogenesis. In contrast, in vivo, bFGF is devoid of effects on neurons in mature hippocampus, raising the possibility that bFGF exhibits developmental stage-specific activity in the complex animal environment. To define neurogenetic effects in the newborn, a single subcutaneous injection of bFGF (20 ng/gm) was administered to postnatal day 1 (P1) rats, and hippocampal DNA content was quantified: bFGF elicited an increase in total DNA throughout adulthood, by 48% at P4, 25% at P22, and 17% at P180, suggesting that bFGF increases hippocampal cell number. To define mechanisms, bromodeoxyuridine (BrdU) was injected at P1 and mitotically labelled cells were assessed at P22: there was a twofold increase in BrdU-positive cells in the dentate granule cell layer (GCL), indicating that bFGF enhanced the generation of neurons, or neuronogenesis, from a cohort of precursors. Moreover, enhanced mitosis and survival led to a 33% increase in absolute GCL neuron number, suggesting that neuron production depends on environmental levels of bFGF. To evaluate this possibility, bFGF-knockout mice were analyzed: hippocampal DNA content was decreased at all ages examined (P3, -42%; P21, -28%; P360, -18%), and total GCL neuron and glial fibrillary acidic protein (GFAP)-positive cell number were decreased by 30%, indicating that bFGF is necessary for normal hippocampal neurogenesis. We conclude that environmental levels of bFGF regulate neonatal hippocampal neurogenesis. As adult hippocampal neuronogenesis was unresponsive to bFGF manipulation in our previous study [Wagner, J.P., Black, I.B. & DiCicco-Bloom, E. (1999) J. Neurosci., 19, 6006], these observations suggest distinct, stage-specific roles of bFGF in the dentate gyrus granule cell lineage. PMID- 11860502 TI - Neurotransmitter release from tottering mice nerve terminals with reduced expression of mutated P- and Q-type Ca2+-channels. AB - Neurotransmitter release is triggered by Ca2+-influx through multiple sub-types of high voltage-activated Ca2+-channels. Tottering mice have a mutation in the alpha1A pore-forming subunit of P- and Q-type Ca2+-channels, two prominent sub types that regulate transmitter release from central nerve terminals. Immunoblotting analysis of purified forebrain terminals from tottering mice revealed an 85% reduction in the protein expression level of the mutated alpha1A subunit compared to expression of the alpha1A subunit in wild-type terminals. In contrast, the expression of the alpha1B subunit of the N-type Ca2+-channels was unchanged. Release of the amino acids glutamate and GABA and of the neuropeptide cholecystokinin (CCK) induced by a short (100 ms) depolarization pulse was unchanged in the terminals of tottering mice. Studies using specific blockers of Ca2+-channels however, revealed a reduced contribution of P- and Q-type Ca2+ channels to glutamate and cholecystokinin release, whereas a greater reliance on N-type Ca2+-channels for release of these transmitters was observed. In contrast, the contribution of the P-, Q- and N-type Ca2+-channels to the release of GABA was not altered in tottering mice. These results indicate that the expression of the alpha1A subunit was decreased in terminals from tottering mice, and that a decreased contribution of P- and Q-type Ca2+-channels to the release of glutamate and cholecystokinin was functionally compensated by an increased contribution of N-type Ca2+-channels. PMID- 11860503 TI - Gelatinase B and TIMP-1 are regulated in a cell- and time-dependent manner in association with neuronal death and glial reactivity after global forebrain ischemia. AB - Matrix metalloproteinases (MMPs) belong to a large family of endopeptidases that regulate the pericellular environment through the cleavage of protein components of the extracellular matrix, membrane receptors and cytokines. MMP activity is controlled by the multifunctional tissue inhibitors of metalloproteinases (TIMPs). Proteases and their inhibitors are critically involved in developmental and pathological processes in numerous organs, including the brain. Global transient cerebral ischemia induces selective delayed neuronal death and neuroinflammation. We compared, in discrete vulnerable and resistant areas of the ischemic rat hippocampus, the kinetics and cellular distribution of gelatinase B and its principal inhibitor TIMP-1 and we assessed by in situ zymography, the net gelatinolytic activity at the cellular level. We show that gelatinases are expressed and active in neurons, suggesting that MMPs play a role in maintaining neural homeostasis. In the ischemic rat brain, expression and activity of gelatinase B, and expression of TIMP-1 are altered in a time-, region- and cell dependent manner. Gelatinase B is induced first in reactive microglia and subsequently in reactive astrocytes. In situ, increases in gelatinase activity accompanied the progression of neuronal death and glial reactivity. Our results suggest that MMPs and TIMPs are involved in cell viability and tissue remodelling in the ischemic brain, and reinforces the idea that the MMP/TIMP system contributes both to neuronal demise and tissue repair in the context of glial reactivity. PMID- 11860504 TI - Insulin-like growth factor-II/Mannose-6-phosphate receptor in the spinal cord and dorsal root ganglia of the adult rat. AB - The insulin-like growth factor-II/mannose-6-phosphate (IGF-II/M6P) receptor is a multifunctional transmembrane glycoprotein, which interacts with a number of molecules, including IGF-II and M6P-containing lysosomal enzymes. The receptor is widely distributed throughout the brain and is known to be involved in lysosomal enzyme trafficking, cell growth, internalization and degradation of IGF-II. In the present study, using autoradiographic, Western blotting and immunocytochemical methods, we provide the first report that IGF-II/M6P receptors are discretely distributed at all major segmental levels of the spinal cord and dorsal root ganglia of the adult rat. In the spinal cord, a high density of [(125)I]IGF-II binding sites was evident in the ventral horn (lamina IX) and in areas around the central canal (lamina X), whereas intermediate grey matter and dorsal horn were associated with moderate receptor levels. The dorsal root ganglia exhibited rather high density of [(125)I]IGF-II binding sites. Interestingly, meninges present around the spinal cord displayed highest density of [(125)I]IGF-II binding compared to any given region of the spinal grey matter or the dorsal root ganglia. Western blot results indicated the presence of the IGF-II/M6P receptor at all major levels of spinal cord and dorsal root ganglia, with little segmental variation. At the cellular level, spinal motorneurons demonstrated the most intense IGF-II/M6P receptor immunoreactivity, followed by interneurons in the intermediate region and deeper dorsal horn. Some scattered IGF-II/M6P immunoreactive fibers were found in the superficial laminae of the dorsal horn and dorsolateral funiculus. The meninges of the spinal cord also seemed to express IGF-II receptor immunoreactivity. In the dorsal root ganglia, receptor immunoreactivity was evident primarily in a subset of neurons of all diameters. These results, taken together, provide anatomical evidence of a role for the IGF-II/M6P receptor in general cellular functions such as transport of lysosomal enzymes and/or internalization followed by clearance of IGF-II in the spinal cord and dorsal root ganglia. PMID- 11860505 TI - Transgenic mice expressing a pH and Cl- sensing yellow-fluorescent protein under the control of a potassium channel promoter. AB - During the last few years a variety of genetically encodable optical probes that monitor physiological parameters such as local pH, Ca2+, Cl-, or transmembrane voltage have been developed. These sensors are based on variants of green fluorescent protein (GFP) and can be synthesized by mammalian cells after transfection with cDNA. To use these sensor proteins in intact brain tissue, specific promoters are needed that drive protein expression at a sufficiently high expression level in distinct neuronal subpopulations. Here we investigated whether the promoter sequence of a particular potassium channel may be useful for this purpose. We produced transgenic mouse lines carrying the gene for enhanced yellow-fluorescent protein (EYFP), a yellow-green pH- and Cl- sensitive variant of GFP, under control of the Kv3.1 K+ channel promoter (pKv3.1). Transgenic mouse lines displayed high levels of EYFP expression, identified by confocal microscopy, in adult cerebellar granule cells, interneurons of the cerebral cortex, and in neurons of hippocampus and thalamus. Furthermore, using living cerebellar slices we demonstrate that expression levels of EYFP are sufficient to report intracellular pH and Cl- concentration using imaging techniques and conditions analogous to those used with conventional ion-sensitive dyes. We conclude that transgenic mice expressing GFP-derived sensors under the control of cell-type specific promoters, provide a unique opportunity for functional characterization of defined subsets of neurons. PMID- 11860506 TI - Control of kinetic properties of GluR2 flop AMPA-type channels: impact of R/G nuclear editing. AB - The GluR2 flop subunit of alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA)-type glutamate receptors greatly determines calcium permeability and kinetic properties of heteromeric AMPA subunit assemblies. Post-transcriptional editing of this subunit at the Q/R/N site controls calcium permeability whereas editing at the R/G site is involved in the regulation of biophysical properties. We used patch-clamp techniques with ultrafast solution exchange to examine the kinetics of recombinant human homomeric GluR2 flop channels transiently expressed in HEK293 cells [edited at the R/G site and Q/R/N site (GR), and unedited (RN) and edited (GN) at the R/G site both with asparagine (N) at the Q/R/N site]. The time constant of desensitization after application of 10 mm glutamate was 1.38 +/ 0.05 ms (n = 10), 5.53 +/- 0.57 ms (n = 7) and 1.33 +/- 0.06 ms (n = 12) for the GluR2 flop GR, RN and GN channels, respectively. The time constant of resensitization was 75 ms for the GluR2 flop RN and 30 ms for the GN channels. The dose-dependence of the peak current amplitude, kinetics of activation and deactivation, and peak open probability did not differ between RN and GN channels. The study shows that desensitization and resensitization kinetics of homomeric GluR2 flop channels are controlled by a single amino acid exchange (glycine by arginine) at the R/G site. Quantitative analysis by computer simulation using a circular kinetic scheme allows the prediction of the main experimental results. PMID- 11860508 TI - Characterizing CGI-94 (comparative gene identification-94) which is down regulated in the hippocampus of early stage Alzheimer's disease brain. AB - The treatment of Alzheimer's disease (AD) remains a major challenge because of the incomplete understanding of the triggering events that lead to the selective neurodegeneration characteristic of AD brains. Here we describe a new protein, CGI-94, that is down-regulated at the mRNA level in the hippocampus of early stage AD brain. Transfection experiments with CGI-94 as a green fluorescent protein (GFP)-fusion-protein show that this protein is translocated into the nucleus of the cell. The finding that this protein, which has a bipartite nuclear localization signal, is also observed in the cytoplasm and extracellular space points to a multifunctional protein. Immunohistochemical analyses reveal that CGI 94 is mainly expressed in neurons of the hippocampal formation and the cortex but not in the cerebellar nucleus. In conclusion, the expression of the nucleolar phosphoprotein CGI-94 appears to be disturbed in early processes of neuronal degeneration. PMID- 11860509 TI - Neuronal expression and regulation of rat inhibitor of apoptosis protein-2 by kainic acid in the rat brain. AB - Inhibitors of apoptosis proteins (IAPs) define a protein family with the ability to counteract cell death by the inhibition of different caspases activated during apoptosis. These proteins are present in different cells, however, the function and roles of IAPs in brain tissue are not fully understood. We report here that RIAP-2, the rat homologue of human cIAP-1/HIAP-2, is expressed in different areas of rat brain as shown by in situ hybridization and immunohistochemistry. Brain regions with relatively high expression of RIAP-2 mRNA included cortex, cerebellum and different subregions of rat hippocampus. Double labelling using a specific anti-RIAP antibody and markers for neurons and glial cells, showed that RIAP-2 is predominantly expressed by nerve cells. Kainic acid treatment, which induces seizures, transiently up-regulated RIAP-2 mRNA levels in cerebral cortex, in the CA1 and dentate gyrus regions of hippocampus, which returned to normal levels at 24 h. However in the CA3 region, RIAP-2 mRNA was decreased at 6 h following an early up-regulation. This region contains neurons particularly vulnerable to kainic acid induced cell degeneration. The decrease in RIAP-2 following kainic acid was also observed using immunohistochemistry. RIAP-2 protein did not colocalize with TUNEL labelling present in cells undergoing cell death. The results show that in the adult rat brain RIAP-2 is expressed mainly by neurons, and that the levels are regulated by kainic acid, which activates glutamate receptors. The decrease in RIAP-2 in specific neuronal populations may contribute to cell degeneration in vulnerable brain regions observed after kainic acid treatment. PMID- 11860510 TI - Macrophage-stimulating protein is a neurotrophic factor for embryonic chicken hypoglossal motoneurons. AB - Macrophage-stimulating protein (MSP) exerts a variety of biological actions on many cell types, but has no known functions in the brain. MSP is structurally related to hepatocyte growth factor (HGF), another pleiotropic factor whose many functions include promoting neuronal survival and growth. To investigate whether MSP is also capable of acting as a neurotrophic factor, we purified hypoglossal motoneurons from the embryonic chicken hindbrain because these neurons are known to express the MSP receptor tyrosine kinase RON. MSP promoted the in vitro survival of these neurons during the period of naturally occurring neuronal death and enhanced the growth of neurites from these neurons. MSP mRNA was detected in the developing tongue whose musculature is innervated by hypoglossal neurons. Our study demonstrates that MSP is a neurotrophic factor for a population of developing motoneurons. PMID- 11860507 TI - Characterization of retinal guanylate cyclase-activating protein 3 (GCAP3) from zebrafish to man. AB - Calmodulin-like neuronal Ca2+-binding proteins (NCBPs) are expressed primarily in neurons and contain a combination of four functional and nonfunctional EF-hand Ca2+-binding motifs. The guanylate cyclase-activating proteins 1-3 (GCAP1-3), the best characterized subgroup of NCBPs, function in the regulation of transmembrane guanylate cyclases 1-2 (GC1-2). The pairing of GCAPs and GCs in vivo depends on cell expression. Therefore, we investigated the expression of these genes in retina using in situ hybridization and immunocytochemistry. Our results demonstrate that GCAP1, GCAP2, GC1 and GC2 are expressed in human rod and cone photoreceptors, while GCAP3 is expressed exclusively in cones. As a consequence of extensive modification, the GCAP3 gene is not expressed in mouse retina. However, this lack of evolutionary conservation appears to be restricted to only some species as we cloned all three GCAPs from teleost (zebrafish) retina and localized them to rod cells, short single cones (GCAP1-2), and all subtypes of cones (GCAP3). Furthermore, sequence comparisons and evolutionary trace analysis coupled with functional testing of the different GCAPs allowed us to identify the key conserved residues that are critical for GCAP structure and function, and to define class-specific residues for the NCBP subfamilies. PMID- 11860511 TI - Voltage-gated ionic currents in an identified modulatory cell type controlling molluscan feeding. AB - An important modulatory cell type, found in all molluscan feeding networks, was investigated using two-electrode voltage- and current-clamp methods. In the cerebral giant cells of Lymnaea, a transient inward Na+ current was identified with activation at -58 +/- 2 mV. It was sensitive to tetrodotoxin only in high concentrations (approximately 50% block at 100 microm), a characteristic of Na+ channels in many molluscan neurons. A much smaller low-threshold persistent Na+ current (activation at < -90 mV) was also identified. Two purely voltage sensitive outward K+ currents were also found: (i) a transient A-current type which was activated at -59 +/- 4 mV and blocked by 4-aminopyridine; (ii) a sustained tetraethylammonium-sensitive delayed rectifier current which was activated at -47 +/- 2 mV. There was also evidence that a third, Ca2+-activated, K+ channel made a contribution to the total outward current. No inwardly rectifying currents were found. Two Ca2+ currents were characterized: (i) a transient low-voltage (-65 +/- 2 mV) activated T-type current, which was blocked in NiCl2 (2 mm) and was completely inactivated at approximately -50 mV; (ii) A sustained high voltage (-40 +/- 1 mV) activated current, which was blocked in CdCl2 (100 microm) but not in omega-conotoxin GVIA (10 microm), omega-agatoxin IVA (500 nm) or nifedipine (10 microm). This current was enhanced in Ba2+ saline. Current-clamp experiments revealed how these different current types could define the membrane potential and firing properties of the cerebral giant cells, which are important in shaping the wide-acting modulatory influence of this neuron on the rest of the feeding network. PMID- 11860513 TI - Dopamine D1/5 receptor stimulation induces c-fos expression in the subthalamic nucleus: possible involvement of local D5 receptors. AB - The activity of neurons in the subthalamic nucleus controls various aspects of movement. The present study examined the action of dopamine receptor agonists on c-fos gene expression in the subthalamic nucleus in normal rats. We found that systemic administration of the dopamine D1/5 receptor agonist, SKF 82958 (1 mg/kg), induces c-fos expression in the subthalamic nucleus. In contrast, systemic administration of the dopamine D2/3 receptor agonist, quinelorane (2 mg/kg) had no effect. When combined, SKF 82958 and quinelorane induced c-fos expression in subthalamic neurons that was similar to that found following administration of SKF 82958 alone. We also examined c-fos expression in the substantia nigra pars reticulata, the major projection area for subthalamic neurons, and found that SKF 82958, but not quinelorane, caused an induction of c fos expression in this area. In order to clarify the mechanisms underlying the SKF 82958-mediated induction of c-fos expression in the subthalamic nucleus and substantia nigra pars reticulata, in situ hybridization for the dopamine D1, D2, D3 and D5 receptor mRNAs was performed. The only significant observation was that D5 receptor mRNA is expressed in subthalamic neurons. The present data show that dopamine, via D1/D5 receptors, upregulates c-fos expression in subthalamic neurons, and that the high expression of D5 receptors in this area might be involved. Taken together, these data suggest that dopamine D1/5 receptors are more important for the action of dopamine in the so-called indirect pathway of the basal ganglia circuitry than what is recognized in current models of basal ganglia organization. PMID- 11860512 TI - Pharmacological validation of behavioural measures of akinesia and dyskinesia in a rat model of Parkinson's disease. AB - In an attempt to define clinically relevant models of akinesia and dyskinesia in 6-hydroxydopamine (6-OHDA)-lesioned rats, we have examined the effects of drugs with high (L-DOPA) vs. low (bromocriptine) dyskinesiogenic potential in Parkinson's disease on three types of motor performance, namely: (i) abnormal involuntary movements (AIMs) (ii) rotational behaviour, and (iii) spontaneous forelimb use (cylinder test). Rats with unilateral 6-OHDA lesions received single daily i.p. injections of L-DOPA or bromocriptine at therapeutic doses. During 3 weeks of treatment, L-DOPA but not bromocriptine induced increasingly severe AIMs affecting the limb, trunk and orofacial region. Rotational behaviour was induced to a much higher extent by bromocriptine than L-DOPA. In the cylinder test, the two drugs initially improved the performance of the parkinsonian limb to a similar extent. However, L-DOPA-treated animals showed declining levels of performance in this test because the drug-induced AIMs interfered with physiological limb use, and gradually replaced all normal motor activities. L DOPA-induced axial, limb and orolingual AIM scores were significantly reduced by the acute administration of compounds that have antidyskinetic efficacy in parkinsonian patients and/or nonhuman primates (-91%, yohimbine 10 mg/kg; -19%, naloxone 4-8 mg/kg; -37%, 5-methoxy 5-N,N-dimethyl-tryptamine 2 mg/kg; -30%, clozapine 8 mg/kg; -50%, amantadine 40 mg/kg). L-DOPA-induced rotation was, however, not affected. The present results demonstrate that 6-OHDA-lesioned rats do exhibit motor deficits that share essential functional similarities with parkinsonian akinesia or dyskinesia. Such deficits can be quantified using novel and relatively simple testing procedures, whereas rotometry cannot discriminate between dyskinetic and antiakinetic effects of antiparkinsonian treatments. PMID- 11860514 TI - Time-dependent inhibition of hippocampal LTP in vitro following contextual fear conditioning in the rat. AB - The effects of contextual fear-learning on hippocampal synaptic excitability were investigated by means of high frequency tetanic stimulation (HFS) in rat brain slices (hippocampal CA1 region), prepared at different intervals (immediately, 24 h or 7 days) after a one-trial contextual fear conditioning paradigm session. In the latter, rats that had previously received aversive electrical footshocks in the experimental apparatus exhibited freezing (the conditioned response) when placed again in the same apparatus (retrieval test). It was shown that contextual fear-learning affects the hippocampal synaptic response. In fact, the HFS produced a decrease in the amplitude of short-term (STP) and long-term potentiation (LTP) when compared to control "naive" subject values. This decrease in STP amplitude could be observed only in slices prepared immediately after the training session. A decrease in the amplitude of long-term but not short-term potentiation was also observed at 24 h. At 7 days, no decreases in amplitude were observed. These modifications may be thought of as specifically associated with the learning process as they were not recorded in brain preparations from "shock only" rats (i.e. those that received the same number of aversive stimuli of equal intensity as the conditioned group but with the shocks compressed temporally so that the shocked subjects could not associate nociceptive stimulation and surroundings - no conditioned freezing during retention testing). In "exploration" preparations (brain slices from rats having only freely explored the experimental apparatus without receiving any adverse stimulation) a decrease in LTP amplitude was recorded only immediately after the training session, and STP was never modified. The synaptic response modifications do not appear to be due to presynaptic events, as they are not associated with paired-pulse facilitation curve (PPF) modifications. The present results show that contextual fear conditioning and exploration of a novel environment (i) reduce the ability to induce synaptic plasticity; (ii) differentially influence STP and LTP and that (iii) the persistence of synaptic modifications depends on an animal's prior experience. PMID- 11860515 TI - Clonidine injections into the lateral nucleus of the amygdala block acquisition and expression of fear-potentiated startle. AB - Numerous studies of aversive learning with different animal models have shown that the noradrenergic system has an important role in the acquisition, consolidation and expression of aversive learning. We used intracerebral clonidine injections to investigate the role of the noradrenergic amygdaloid system in the fear-potentiated startle paradigm. Clonidine is a noradrenergic alpha2-receptor agonist which can decrease noradrenergic transmission by stimulating presynaptic alpha2-receptors. Rats received injections of 0, 2.5, 5 and 10 nmol clonidine into the lateral amygdala (i) before fear-conditioning, (ii) immediately after fear-conditioning, (iii) before testing and (iv) before both fear-conditioning and the testing of conditioned fear. Clonidine injections blocked the acquisition and expression of conditioned fear. The effect on acquisition was not caused by state-dependency or possible side-effects of clonidine on consolidation. Given that clonidine decreases amygdaloid noradrenaline release, these results show a crucial role of noradrenergic transmission within the amygdala in classical fear-conditioning. Surprisingly, both the acquisition and the expression of conditioned fear were blocked after amygdaloid injections of clonidine, suggesting that amygdaloid noradrenaline is necessary to induce both unconditioned and conditioned fear. PMID- 11860516 TI - Absence of dopamine D4 receptors results in enhanced reactivity to unconditioned, but not conditioned, fear. AB - The prefrontal cortex receives a major dopaminergic input from the ventral tegmental area, which plays an important role in the integration of neuronal signals influencing behavioural responses to stressful environmental stimuli. The dopamine D4 receptor (D4R) is expressed at highest levels in the prefrontal cortex and is the predominant D2-like receptor localized in this brain area. To investigate the functional significance of D4Rs in dopamine-mediated responses we have analysed a strain of mice lacking this receptor subtype (Drd4-/-). Wild-type and Drd4-/- mice were challenged in two different approach/avoidance conflict paradigms: the elevated plus maze and the light/dark preference exploration test. By these behavioural measures Drd4-/- mice showed heightened avoidance to the more fear-provoking areas of each maze as demonstrated by reduced exploration of the open arms of the plus maze and longer latencies to explore the illuminated compartment of the light/dark shuttle box. These exaggerated avoidance behaviours were further enhanced by an additional handling stress but completely prevented by anxiolytic agents such as the benzodiazepine midazolam and ethanol. Although Drd4-/- mice displayed heightened anxiety, they exhibited normal ethanol preference and consumption in a two-bottle choice test. Learned fear responses evaluated by contextual, cued and instrumental fear-conditioning tests showed no difference between wild-type and Drd4-/- mice. Taken together these results indicate that the absence of D4Rs increases avoidance behaviour to unconditioned stimuli and does not impair behavioural reactions to Pavlovian fear-conditioning, suggesting that the D4R could play a key role in the dopaminergic modulation of cortical signals triggered by environmental stimuli. PMID- 11860517 TI - The neural substrate of orientation short-term memory and resistance to distractor items. AB - We used Positron Emission Tomography to map the neural substrate of human short term memory for orientation, defined as retaining a single orientation in memory over a long delay, by comparing a successive discrimination task with a 6-s delay to the same task with a brief 0.3 s delay and to an identification control task. Short-term memory engaged the superior parietal lobe bilaterally, the middle occipital gyrus bilaterally and the left dorsolateral prefrontal cortex. In addition, we studied the resistance to a distractor item by comparing the successive discrimination task with long delay, with and without an intervening distractor stimulus. This manipulative process engaged left ventral premotor cortex and left dorsolateral prefrontal cortex. The activation of left dorsolateral prefrontal cortex is interpreted as reflecting co-ordination between task components. These results, combined with those of two previous studies using an identical reduction strategy, underscore the functional heterogeneity in the prefrontal cortex during short-term and working memory. PMID- 11860518 TI - Substance P in the descending cholinergic projection to REM sleep-induction regions of the rat pontine reticular formation: anatomical and electrophysiological analyses. AB - Release of acetylcholine within the pontine reticular formation (PRF) from the axon terminals of mesopontine cholinergic neurons has long been hypothesized to play an important role in rapid eye movement (REM) sleep generation. As some of these cholinergic neurons are known to contain substance P (SP), we used anatomical, electrophysiological and pharmacological techniques to characterize this projection in the rat. Double immunofluorescence demonstrated that 16% of all cholinergic neurons within the mesopontine tegmentum contained SP; this percentage increased to 27% in its caudal regions. When double immunofluorescence was combined with retrograde tracing techniques, it was observed that up to 11% of all SP-containing cholinergic neurons project to the PRF. Whole-cell patch clamp recordings from in vitro brainstem slices revealed that SP administration depolarized or evoked an inward current in a dose-dependent manner in all PRF neurons examined, and that these effects were antagonized by a SP antagonist. The amplitude of the SP-induced inward current varied with changes in the Na+ concentration, did not reverse at the calculated K+ or Cl- equilibrium potentials, and was not attenuated in the presence of tetrodotoxin, low Ca2+ concentration or caesium ions. These data suggest that activation of a tetrodotoxin-insensitive cation channel(s) permeable to Na+ is responsible for a SP-induced inward current at resting membrane potentials. The depolarizing actions of SP appeared to be primarily due to activation of the adenylate cyclase pathway, and were additive with cholinergic receptor activation even at maximal concentrations. These data indicate that SP is colocalized in a subpopulation of mesopontine tegmental cholinergic neurons projecting to REM sleep-induction regions of the PRF, and that actions of these two neuroactive substances on PRF neurons are additive. If SP is coreleased with acetylcholine, the additive actions of the two neurotransmitters might heighten the excitability of postsynaptic PRF neurons and ensure the initiation and maintenance of REM sleep. PMID- 11860519 TI - Participation of the homing pigeon thalamofugal visual pathway in sun-compass associative learning. AB - The ascending thalamofugal visual pathway in pigeons (Columba livia) terminates in the telencephalic wulst. Characterizing the role of this pathway in visually guided behaviour has remained a challenge. To determine whether this pathway, and in particular the wulst, may participate in sun-compass-guided behaviour in homing pigeons, intact, ectostriatum-lesioned or wulst-lesioned pigeons were trained to use their sun compass to locate the direction of a food reward in an outdoor, octagonal arena. Control and ectostriatum-lesioned pigeons learned the task well, and orientated appropriately during the first trial of the last three training sessions and after a phase-shift manipulation. In contrast, the wulst lesioned pigeons learned the task but they took more sessions to learn, and their directional choices were more scattered during the first trial of the last three training sessions and after the phase-shift manipulation. A subsequent regression analysis indicated that deeper layers of the wulst might have made more of a contribution to the observed behavioural impairments. The data indicate that the homing pigeon wulst participates in visually guided behaviour when the sun compass is used to learn the directional location of a goal. PMID- 11860520 TI - Expression of CRABP I mRNA in fastigial cells of the developing cerebellum. AB - The expression of the cellular retinoic acid binding protein type I (CRABP I) was examined in the early phase of cerebellar development in the mouse. The CRABP I was expressed from embryonic day (E) 10.5 to E15.5 in the cerebellar plate. The expression was diffused at E10.5-E11.5 and thereafter localized in a small rostrodorsal area of the cerebellar territory of both sides. By using in situ hybridization and both immunohistochemistry and carbocyanine tracing procedures, we identified the fastigial cells as the population that expresses CRABP I in the cerebellum. The results suggest that these cells play a critical role in the early development of the cerebellum. PMID- 11860521 TI - Opposing actions of neuropeptide Y and light on the expression of circadian clock genes in the mouse suprachiasmatic nuclei. AB - The circadian clockwork of the hypothalamic suprachiasmatic nuclei (SCN) is synchronized by light and by nonphotic cues. The core timing mechanism is cell autonomous, based on an autoregulatory transcriptional/translational feedback loop of circadian genes and their products. This study investigated the effects of neuropeptide Y (NPY), a potent nonphotic resetting cue, and its interaction with light in regulating clock gene expression in the SCN in vivo. Injection of NPY adjacent to the SCN and transfer to darkness 7 h before scheduled lights out, shifted the circadian activity-rest cycle. Exposure to light for 1 h immediately after NPY infusion blocked this behavioural response. NPY-induced shifts were accompanied by suppression of both mPer1 and mPer2 mRNA in the SCN, assessed 3 h after infusion. mPer mRNAs were not altered 1 h after infusion. Levels of mClock mRNA or mCLOCK immunoreactivity in the SCN were not affected by NPY at either time point. In parallel to the behavioural response, the NPY-induced suppression of mPer genes in the SCN was attenuated when a light pulse was delivered immediately after the infusion. These results identify mPer1 and mPer2 as molecular targets for both photic and nonphotic (NPY-induced) resetting of the clockwork, and support a synthetic model of circadian entrainment based upon convergent up- and downregulation of mPer expression. PMID- 11860523 TI - Teaching pharmacology together in Europe. PMID- 11860524 TI - Metabolism of methoxymorpholino-doxorubicin in rat, dog and monkey liver microsomes: comparison with human microsomes. AB - The morpholino anthracycline, methoxymorpholino-doxorubicin (MMDx) is a novel anticancer agent. The metabolism of this highly lipophilic doxorubicin analogue is not fully elucidated. MMDx is metabolically activated in vivo, resulting in an 80-fold increase in potency over the parent drug. In this study, MMDx in vitro metabolism was compared in rat, dog, monkey and human liver microsomes. When microsomal fractions were incubated with MMDx, 6-8 metabolites were formed depending on the species and on the substrate concentrations. Among these eight metabolites, three comigrated with authentic standards, namely MMDx-ol, PNU156686 and PNU159682, and the five others are in the process of being characterized. Quantitatively, monkey and human metabolize MMDx with a higher rate than rat and dog. Qualitatively, MMDx metabolic profile in dog microsomes was different from the three other species. MMDx-ol was predominant in dog and only minor in other species. In conclusion, MMDx metabolism was species-different. Rat and monkey liver microsomes may be used as models to study MMDx metabolism in humans. Dog liver microsomes may be a good model for studying the formation of MMDx-ol. PMID- 11860525 TI - Effects of endothelin-A receptor antagonism on bilateral renal function in renovascular hypertensive rats. AB - Recent studies indicated an enhanced expression of Endothelin (ET) in the kidney contralateral to the vascular clip in two-kidney, one-clip (2K-1C) Goldblatt hypertension. We proposed that the enhanced intrarenal ET production might be responsible for altered haemodynamic and excretory capability of the unclipped kidney (UK) of 2K-1C renovascular hypertensive rats. Therefore, we examined the changes in arterial pressure and split renal function in the clipped (CK) and UK simultaneously, in response to chronic administration of the selective ETA receptor blocker (A-127722), given orally at a dose of 30 mg/kg/day for 3 weeks starting from the beginning of the 4th week of clipping. Systolic pressure averaged 177 +/- 7 mmHg in control rats (n=15) and 164 +/- 9 mmHg in treated rats (n=16) and the difference was not statistically different. Glomerular filtration rate (GFR), renal plasma flow (RPF) and renal vascular resistance (RVR) in the UK were not different between control and treated groups. Data were then analyzed by classifying rats as moderate hypertensives (MAP < 180 mmHg), and severe hypertensives (MAP > 180 mmHg). In the moderately hypertensive group, average MAP was 143 +/- 5 mmHg and 138 +/- 4 mmHg in control (n=9) and treated (n=10) groups, respectively. In the severely hypertensive group, average MAP was 192 +/- 5 mmHg and 188 +/- 5 mmHg in control (n=6) and treated (n=6) groups, respectively. GFR and RPF were significantly improved in the UK of only the severely hypertensives who received the antagonist. However, the ET(A) antagonist blunted the sodium loss in both CK and UK of severely hypertensive rats. We conclude that ET(A) receptors do not play a role in the progression of hypertension in 2K-1C renovascular hypertensive rats. Yet, ET(A) receptors play an important role in altering renal hemodynamics of the unclipped kidneys in severe degrees of renovascular hypertension. PMID- 11860526 TI - Deleterious effects of nifedipine on smooth muscle cells implies alterations of intracellular calcium signaling. AB - Nifedipine (NIF), a calcium channel blocker (CCB) from the first generation of dihydropyridines, induces detrimental effects on patients with cardiovascular diseases. We designed experiments to study, at cellular and molecular level, the mechanisms involved in the induction of deleterious effects by this drug. To this purpose, cultured human smooth muscle cells (HSMC) were used. The effect of NIF and two other CCB (FEL, AML) and inhibitors of intracellular signaling pathways (RR, TG, CAF and GEN) on intracellular calcium [Ca(2+)]I was determined by spectrofluorimetry using Fura 2 AM assay. The results showed that: (i) 10 microM NIF induced the increase of [Ca(2+)]I above the basal values (202.77 +/- 23.98 nM vs. 48.68 +/- 6.45 nM), an effect that was prevented by RR (50.45 +/- 13.9 nM) and was not induced by the two other CCB; (ii) NIF had a thapsigargin-like effect, because it induced the same release of intracellular calcium as TG (212.1 +/- 25.62 nM); (iii) The response to NIF was reduced by 40% after the inhibition of IP3 receptor (121.21 +/- 26.01 nM) and by 50% after the inhibition of tyrosine kinase (101.91 +/- 7.76 nM). Together, these data demonstrate that NIF produces a deregulation of intracellular calcium homeostasis. The abnormal increase of [Ca(2+)]I is due to the activation of store operated channels from the plasma membrane responsible for capacitative calcium entry, a process modulated by the activity of tyrosine kinase and the Ca(2+)-ATPase pump from the sarcoplasmic reticulum. PMID- 11860527 TI - Modulation of dopamine release in the guinea-pig retina by G(i)- but not by G(s)- or G(q)-protein-coupled receptors. AB - The modulation of dopamine release from the guinea-pig retina was studied using maximally effective concentrations of 10 agonists acting on G(i)-, G(s)- or G(q) protein-coupled receptors (PCRs). Retinal discs were preincubated with [(3)H]noradrenaline and superfused; tritium overflow was evoked electrically. The following compounds acting on G(i)-PCRs reduced the tritium overflow, which represents quasi-physiological dopamine release under the experimental conditions of our study: the dopamine and alpha(2)-adrenoceptor agonist B-HT 920 by 95%, the muscarinic agonist oxotremorine by 96%, melatonin by 94%, the cannabinoid agonist WIN 55,212-2 by 71% and histamine by 66%. Tritium overflow was not affected by serotonin or by agonists acting on G(s)-PCRs (ACTH1-24 and the beta-adrenoceptor agonist procaterol) and G(q)-PCRs (angiotensin II and bradykinin). The effects of B-HT 920, oxotremorine and melatonin were studied in more detail using appropriate antagonists. The inhibitory effect of a submaximally active concentration of B-HT 920 was counteracted by the dopamine D(2/3) antagonist haloperidol but not affected by the alpha(2)-adrenoceptor antagonist phentolamine. The muscarinic antagonist atropine shifted to the right the concentration-response curve of oxotremorine (pA(2) 8.7) and the melatonin MT(2) antagonist 4-P-PDOT produced a rightward shift of the concentration-response curve of melatonin (pA(2) 10.6). Melatonin was also studied in superfused brain slices (from the guinea-pig) preincubated with [(3)H]noradrenaline. The electrically evoked tritium overflow in cerebrocortical, hippocampal and hypothalamic slices (representing quasi-physiological noradrenaline release) and in striatal slices (representing quasi-physiological dopamine release) was not affected by melatonin at a concentration that causes the maximum effect in retinal discs. In conclusion, dopamine release in the guinea-pig retina is inhibited via G(i)-PCRs including dopamine (D(2/3)), muscarinic and melatonin (MT(2)) receptors but not affected via any of the G(s)- or G(q)-PCRs under study. Unlike in the retina, melatonin fails to inhibit monoamine release in four brain regions of the guinea-pig. PMID- 11860528 TI - Serum uric acid levels: a useful but not absolute marker of compliance with fenofibrate treatment. AB - The purpose of our study was to investigate whether measurement of serum uric acid levels is a reliable method to assess adherence to fenofibrate treatment. This was a 21 week, open-label study conducted in our lipid clinic. After an 8 week dietary baseline phase, we implemented a treatment phase, during which patients received 200 mg/day micronized fenofibrate for 3 months. Serum lipid profiles, including levels of lipoprotein(a) and fibrinogen, serum uric acid, as well as muscle and liver enzymes, were measured after the dietary phase, and at the end of the 3 month treatment period. Compliance was assessed using a clinical interview. A significant decrease in serum uric acid levels was observed in the compliant, while no significant change was noticed in the non-compliant. The compliant had lower serum uric acid concentrations compared to the non-compliant (P < 0.01) after fenofibrate treatment. Furthermore, they demonstrated lower levels of triglycerides and higher high-density lipoprotein (HDL) cholesterol concentrations (P < 0.05 for both parameters). The results show that serum uric acid concentrations may be used as a useful tool to assess compliance with fenofibrate therapy. PMID- 11860529 TI - Clinical pharmacology and therapeutic use of the new antiepileptic drugs. AB - Although older generation antiepileptic drugs (AEDs) such as carbamazepine, phenytoin and valproic acid continue to be widely used in the treatment of epilepsy, these drugs have important shortcomings such as a highly variable and nonlinear pharmacokinetics, a narrow therapeutic index, suboptimal response rates, and a propensity to cause significant adverse effects and drug interactions. In an attempt to overcome these problems, a new generation of AEDs has been introduced in the last decade. Compared with older agents, some of these drugs offer appreciable advantages in terms of less variable kinetics and, particularly in the case of gabapentin, levetiracetam and vigabatrin, a lower interaction potential. Lamotrigine, topiramate, zonisamide and felbamate protect against partial seizures and a variety of generalized seizure types, vigabatrin is effective against partial seizures (with or without secondary generalization) and infantile spasms, while the use of oxcarbazepine, tiagabine and gabapentin is mainly restricted to patients with partial epilepsy (and, in the case of oxcarbazepine, also primarily generalized tonic-clonic seizures). Levetiracetam, the latest AED to be introduced, has been found to be effective in partial seizures, but its potentially broader efficacy spectrum remains to be determined in clinical studies. Currently, the main use of new generation AEDs is in the adjunctive therapy of patients refractory to older agents. However, due to advantages in terms of tolerability and ease of use, some of these drugs are increasingly used for first-line management in certain subgroups of patients. Due to serious toxicity risks, felbamate and vigabatrin should be prescribed only in patients refractory to other drugs. In the case of vigabatrin, however, first line use may be justified in infants with spasms. PMID- 11860530 TI - Pharmacokinetic-pharmacodynamic modeling of the inhibitory effect of erythromycin on tumour necrosis factor-alpha and interleukin-6 production. AB - Erythromycin inhibits the production of tumour necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL6) induced by heat-killed Streptococcus pneumoniae in human whole blood ex-vivo. The objective of the present study was to determine and characterize the concentration-effect relationship of this phenomenon in order to predict its possible clinical relevance. Six healthy volunteers received a single intravenous dose of 1000 mg erythromycin. Blood samples were obtained up to 4 h after drug administration. Samples were assayed for erythromycin concentrations and (after heat-killed Streptococcus pneumoniae stimulation) for TNF-alpha and IL6 concentrations. Effect vs. time data from individual subjects were fitted to the indirect response model with an Emax concentration-effect relationship. Simulations of these effects were performed for therapeutic intravenous and oral erythromycin dosage regimens. The geometric means of the values of Kin, Kout and EC50 were 15.4 microg/h, 0.82/h, 9.4 mg/L for TNF-alpha and 321 microg/h, 2.02/h, 18.3 mg/L for IL6. Simulations revealed a maximal inhibition of TNF-alpha concentrations of 35%, 50%, 16% and 27% at erythromycin dosages of 500 mg i.v., 1000 mg i.v., 500 mg p.o and 1000 mg p.o. q 6 h, respectively, whereas a maximal inhibition of IL6 of 29%, 44%, 13% and 22% are predicted for the respective regimens. The inhibitory effect of erythromycin on TNF-alpha and IL6 production can be adequately described by the indirect response model with an Emax concentration-effect relationship. Simulations predicted a substantial decrease of production of these cytokines at intravenous and to a much lesser extent at oral erythromycin dosage regimens. PMID- 11860531 TI - Immunology of the peritoneal cavity: relevance for host-tumor relation. AB - The peritoneal membrane, formed by a single layer of mesothelial cells, lines the largest cavity of the human body. Anatomic structures of the peritoneal cavity, along with resident leukocyte populations, play an important role in the defense against microorganisms invading by breaching the gut integrity or ascending through the female genital tract. Local immune mechanisms in the peritoneal cavity are also important in patients undergoing peritoneal dialysis and in women with endometriosis. There is now extensive evidence demonstrating the significance of peritoneal immune mechanisms in the control of metastatic spread. Leukocytes belonging to both the innate and adaptive immune systems are present in the peritoneal cavity of normal subjects as well as in patients with intra abdominal cancer. There is now increased understanding of the mechanisms that not only allow the tumor cells to escape the detection and destruction by the host immune system, but also to use the inflammatory mechanisms to promote tumor growth and spread inside the peritoneal cavity. Malignant ascites represents a model for the study of the interaction between tumor cells and the host immune system as well for the analysis of the tumor microenviroment. The peritoneal immune system may be stimulated by intraperitoneal administration of biologic agents. This peritoneal immunotherapy may be used for palliation of malignant ascites, or as a consolidation strategy in patients with minimal residual disease. PMID- 11860532 TI - Laparoscopic lower para-aortic staging lymphadenectomy in stage IB2, II, and III cervical cancer. AB - The presence of metastases in the para-aortic lymph nodes has important implications in the management of cervical carcinoma in many centers. In this study we investigated the role of laparoscopic lower para-aortic lymphadenectomy in patients with cervical carcinoma. In 42 consecutive patients with stage IB2 IIIB cervical carcinoma without suspicious para-aortic lymph nodes on CT scanning, a laparoscopic lower para-aortic lymphadenectomy was performed between January 1998 and April 2001. The transperitoneal route was used in the first 21 patients, and in the remaining 21 patients the procedure was started with the left retroperitoneal approach. The number of patients with stage IB2, II, and III was 7, 30, and 5, respectively. The median age was 51 years (range 30-81). The median weight and length were 62 kg (range 45-83) and 162 cm (range 150-175), respectively. In four patients-two operated via the transperitoneal route and the other two operated via the retroperitoneal route-the procedure was abandoned, in three patients because of adhesions following prior surgery and in one because of a camera failure. Conversion from the retropertitoneal to the transperitoneal approach was necessary because of a peritoneal tear in five of the 21 cases (all of them in the first 10 cases). Median estimated blood loss during the operation was 78 ml (range 10-300). The median hemoglobin (Hb) decrease was 1.3 g/dL (range 0-3.7). The median duration of the procedure was 64 min (range 20-115). Lymphadenectomy was stopped when one of the nodes was positive on frozen section. The number of para-aortic lymph nodes removed ranged from one to 15 (median 6). In seven (18%) of the 38 patients with successful lymphadenectomy, para-aortic metastases were observed (stage IB2, 1/6; stage II, 5/28 and stage III, 1/4). The only major complication was the development of a retroperitoneal hematoma on the first postoperative day in one patient (Hb fall of 3.7 g/dL). Patients with para aortic lymph node metastases were treated with extended field para-aortic radiotherapy. None of these developed bowel obstruction. The median follow-up was 15 months (range 1-40 months). Actuarial 1-year crude survival was 88% and 33%, respectively, for patients with negative and positive para-aortic nodes. Laparoscopic para-aortic staging in cervical carcinoma is feasible with low morbidity. Eighteen percent of the patients, without suspicion of para-aortic metastases on CT, proved to have metastases in the lower para-aortic region. PMID- 11860533 TI - High-dose-rate brachytherapy for vaginal cancer: learning from treatment complications. AB - Historically, early stage vaginal cancer has been treated with low-dose-rate (LDR) brachytherapy with or without external beam radiation therapy (EBRT). Complication rates have been low and treatment efficacious. Although high-dose rate (HDR) brachytherapy has been used for cervical cancer in many countries for over a decade, only more recently has it been integrated into treatment plans for vaginal cancer. This paper describes three patients treated with HDR brachytherapy who experienced significant late effects. Given the very limited amount of literature regarding the use of HDR brachytherapy in vaginal cancer, this analysis potentially contributes to an understanding of treatment-related risk factors for complications among patients treated with this modality.A focused review of hospital and departmental treatment records was done on three patients treated with HDR brachytherapy. Abstracted information included clinical data, treatment parameters (technique, doses, volume, combinations with other treatments) and outcomes (local control, survival, early and late effects). A review of the available literature was also undertaken. All patients had significant complications. Although statistical correlations between treatment parameters and complications are impossible given the limited number of patients, this descriptive analysis suggests that vaginal length treated with HDR brachytherapy is a risk factor for early and late effects, that the distal vagina has a lower radiation tolerance than the upper vagina with HDR as in LDR, and that combining HDR with LDR as done in our experience carries a high risk of late toxicity. Integration of HDR brachytherapy techniques into treatment plans for early stage vaginal cancers must be done cautiously. The etiology of the significant side effects seen here is likely to be multifactorial. For users of HDR brachytherapy in vaginal cancer, there is a need to further refine and standardize treatment concepts and treatment delivery. Ideally this will be based on continued careful observation and reporting of both favorable and unfavorable outcomes and experiences. PMID- 11860534 TI - The prognostic value of histopathologic grading parameters and microvessel density in patients with early squamous cell carcinoma of the uterine cervix. AB - The purpose of this study was to investigate the prognostic importance of clinical and histopathologic factors, including malignancy grading systems (MGS), partial index (PI), invasive front grading (IFG), and microvessel density. A complete geographic series of 172 early stage (FIGO I-II) cervical carcinomas treated by Wertheim-Meigs surgery during the period 1965-1990 was studied. The patients were followed up for at least 10 years. Significant prognostic factors for disease-free survival were lymph node status (P < 0.0000001), radical surgical margins (P = 0.00003), and tumor size (P = 0.008). In a multivariate Cox analysis it was shown that lymph node status was the single most important prognostic factor with regard to disease-free survival. The total MGS and the PI scores were highly significantly (P = 0.0001) associated with pelvic lymph node metastases and disease-free survival rate in squamous cell carcinomas. The MGS and the PI systems were superior to the IFG system in predicting lymph node metastases. The total IFG score was also a statistically highly significant (P = 0.003) prognostic factor with regard to disease-free survival in both univariate and multivariate analyses. Microvessel density was a nonsignificant prognostic factor. There was a highly significant (P = 0.002) association between vascular space invasion of tumor cells and the presence of lymph node metastases. In conclusion, histopathologic malignancy grading systems provide valuable prognostic information in patients with early stage squamous cell carcinomas of the uterine cervix. PMID- 11860535 TI - Clinicopathological study of the pattern and significance of cervical involvement in cases of endometrial adenocarcinoma. AB - The pattern of cervical involvement in 107 endometrial adenocarcinomas was assessed. The cervix was involved in 29%, higher than noted in previous studies. In 40.6%, the lesion was confined to surface endocervical epithelium only; the remainder had cervical stromal involvement. In the majority only small areas within the circumference of the cervix were affected, indicating a need for adequate tissue sampling. In some cases, malignant epithelium was found as a "migrant" within the endocervical canal, entrapped within cervical mucus or applied to surface epithelium, supporting the concept that endometrial cancer spreads by surface contiguity or implantation rather than by deep tissue planes or via lymphatic channels. Our findings reinforce the view that high-grade lesions and histological subtypes such as uterine serous papillary carcinoma are associated with a later presentation, higher stage and poorer prognosis. We have identified atypical changes in endocervical epithelium that may be misinterpreted as cervical involvement, particularly in the form of atypical reserve cell hyperplasia with a micropapillary pattern that may reflect a reaction to the presence of tumor. It is our assertion that the presence of tumor "migrants", and not endocervical surface atypia, is an indicator of increased probability of cervical involvement by endometrial adenocarcinoma (P = 0.015). PMID- 11860536 TI - Laparoscopically assisted vaginal hysterectomy versus abdominal hysterectomy in stage I endometrial cancer. AB - The purpose of this study was to evaluate and compare laparoscopic treatment for stage I endometrial cancer with the traditional transabdominal approach. From July 1996 to July 1998, 61 patients with clinical stage I endometrial cancer were treated at the Gynaecology Oncology Unit at the Royal North Shore of Sydney, Australia. Twenty-nine patients were treated with laparoscopic assisted vaginal hysterectomy (LAVH) and bilateral salpingo-oophrectomy (BSO) plus minus laparoscopic pelvic lymphadenectomy (LPLA), while 32 patients were treated with the traditional laparotomy and underwent total abdominal hysterectomy (TAH) and BSO plus minus pelvic lymphadenectomy (PLA). The main outcomes studied were operative time, blood loss, blood transfusion, intraoperative complications, postoperative complications, duration of hospital stay, and number of lymph nodes obtained. In conclusion, laparoscopic treatment of endometrial cancer is safe in the hands of experienced operators with minimal intraoperative and postoperative complications. This procedure is associated with significantly less blood loss and shorter hospitalization; however, it is associated with significantly longer operating time. Proper selection of patients for the laparoscopic procedure is the vital step in achieving the major goals of this approach. PMID- 11860537 TI - Immunohistochemical localization of metallothionein in benign and malignant epithelial ovarian tumors. AB - Metallothioneins (MTs) are a group of low-molecular-weight proteins that are overexpressed in a variety of human neoplasms and are related to differentiation and prognosis in some tumor types. This study investigated immunohistochemically detectable metallothionein expression in benign and malignant ovarian surface epithelial tumors of serous, mucinous, and endometrioid types. MT expression was observed in 56% of carcinomas (n = 139) and in 2% of benign neoplasms (n = 81). Of the malignant tumors, MT expression was found in 68% of endometrioid, 56% of mucinous, and 52% of serous neoplasms. There was increased MT expression in grade 3 carcinomas (64%) as compared with grade 2 (60%) and grade 1 (23%). The overexpression of MT in malignant as opposed to benign ovarian surface epithelial tumors may suggest a role in tumorigenesis. Analogous to the situation in endometrial carcinomas, there is a tendency toward higher expression in poorly differentiated tumors. Whether high MT expression is an independent prognostic factor and increased expression indicates chemotherapy resistance in ovarian cancer, as has been previously suggested, should be determined by further studies. PMID- 11860539 TI - Immunohistochemical study of p185 HER2 and DF3 in primary breast cancer and correlation with CA-15-3 serum tumor marker. AB - Human epidermal growth factor receptor 2 (p185 HER2) oncoprotein immunohistochemical expression and DF3 antigen distribution were evaluated in 129 patients with primary breast cancer. p185 HER2 overexpession was positively correlated with the degree of differentiation, metastatic disease, progesterone receptors, and cytoplasmic distribution of DF3 antigen. p185 HER2 overexpression had prognostic significance for the disease-free interval. PMID- 11860538 TI - Histologic and immunohistochemical analysis of tissue response to adenovirus mediated herpes simplex thymidine kinase gene therapy of ovarian cancer. AB - Herpes simplex virus (HSV) thymidine kinase (tk) gene incorporated into adenovirus was delivered intraperitoneally (ip) followed by an antiherpetic prodrug and topotecan in patients with recurrent epithelial ovarian cancer. Tissue response was evaluated. Ten patients underwent secondary debulking with subsequent delivery of ADV-HSV-tk therapy. Two patients each were treated at dose level 1 (2 x 10(10) vector particles = VP), 2 (2 x 10(11) VP), and 3 (2 x 10(12) VP); four patients were treated at dose level 4 (2 x 10(13) VP). Five patients underwent second-look surgery about one month after gene therapy (GT). Treatment response, presence of vector DNA, protein expression of steroid hormone receptors, p53, c-erbB2 and Ki67 protein were analyzed. At second-look, two out of five patients were tumor-free and none of their peritoneal biopsies showed vector DNA. After GT, the vital tumor mass was smaller, desmoplastic reaction had increased, and tumors were less differentiated with an increase of Ki67 expression. There was no change in expression of hormone receptors, p53, or c erbB2. ADV-HSV-tk GT appears to eliminate cells with higher differentiation first and might induce fibrosis. Dedifferentiation might render residual cells more sensitive to chemotherapy secondary to their subsequent higher mitotic activity. PMID- 11860540 TI - The experimental study of ovarian carcinoma vaccine modified by human B7-1 and IFN-gamma genes. AB - With the advance of immunology, immunogene therapy is becoming a possible therapeutic alternative to advanced cancer. The aim of tumor immunogene therapy is to enhance the immune response to tumors. Evidence suggests that CD80 (B7-1) and Interferon-gamma (IFN-gamma) play important roles in antitumor immunity induced by T lymphocyte. To study the antitumor immune effects of ovarian carcinoma vaccine modified with human B7-1 and IFN-gamma genes, we constructed the bicistronic retroviral vector pLXSN, encoding human B7-1 and IFN-gamma. In vitro, the primary ovarian carcinoma cells were transduced with the retroviral vector and prepared as a vaccine. Then autologous lymphocytes were irritated with the ovarian carcinoma cells for competition inhibitory cytotoxic testing. The tumor-specific cytotoxic activity was greatly enhanced by the double gene modified vaccine. In vivo, the tumorigenicity of the double gene-modified ovarian cell line 3AO/B7-1.IFN-gamma was evaluated in a human immune reconstituted SCID mice (hu-PBL-SCID) model. The double-gene modification markedly decreases tumor genecity. Together, the results suggest that combining B7-1 and IFN-gamma could be a useful therapeutic approach in ovarian cancer. PMID- 11860541 TI - Use of CA-125 and ultrasound in high-risk women. AB - Our objective was a retrospective study reporting on ovarian cancer screening in a high-risk female population using both CA-125 and ultrasound over a 7-year period. We used risk estimates of carrying a BRCA mutation that were based on family history. Subjects were screened with CA-125 and ultrasound every 6 months. Each of 311 high-risk subjects had between 1 and 17 screening visits. Overall, 33 of 1209 (2.7%) CA-125 results were abnormal (>35 U/ml); 226 of 1342 (17%) ultrasounds were abnormal, with abnormalities ranging from benign appearing cystic changes to more ominous patterns. Since entry into the program, 29 subjects (9%) have undergone surgery. In 20 of these, the preoperative screening was normal; in six, only the ultrasound was abnormal, and in two, only the CA-125 was abnormal (46-91 U/ml). In only one subject undergoing surgery were both serial CA-125 levels (52-91 U/ml) and ultrasound abnormal. In 7 years of screening, one patient (0.3%) has been diagnosed with ovarian cancer (stage IA, grade 1 endometrioid adenocarcinoma). Overall, 31 (10%) subjects have completed BRCA testing. We conclude that despite screening results comparable to other studies, the detection of only one ovarian cancer over 7 years is lower than expected. Explanations for this observation are discussed. Despite the limitations of CA-125 and ultrasound, we continue to recommend these screening modalities for high-risk women. At the present time, they offer the best opportunity to detect ovarian cancers early. With increasing knowledge of BRCA testing, more women may benefit from this testing in assessing their personal risk. PMID- 11860542 TI - Characterization of mast cells according to their content of tryptase and chymase in normal and neoplastic human uterine cervix. AB - Mast cells (MC) have been associated with diverse human cancers. The primary function of these cells is to store and release a number of biologically active mediators, including the serine proteases tryptase and chymase. These proteases have been closely related with angiogenesis and tumor invasion, two critical steps during tumor progression. In the present work we analyzed the presence of MC in human uterine cervix from both normal and neoplastic tissues by using metachromatic, immunohistochemical, and enzymohistochemical staining. Tryptase positive (MCT)- and tryptase/chymase-positive (MCTC)-mast cells were found in both normal and neoplastic tissues. The phenotype predominantly expressed in normal tissues as well as in benign and malignant lesions of the uterine cervix was the MCT. The total number of MC remained constant through the different stages of malignant transformation (cervical intraepithelial neoplasia grade 1-3) but a significant increase in the invasive carcinoma (IC) group was observed, this increase being mainly due to the MCT phenotype. Furthermore, we detected abundant MCT but not MCTC infiltrating tumors in sections of IC. Regarding the potent angiogenic properties described for tryptase, these findings suggest that in advanced stages of malignancy the significant number of MCT distributed within the cervical tissues could provide an effective mechanism to create the abundantly vascularized microenvironment required for tumor cells to proliferate and disseminate. PMID- 11860543 TI - Sequence variations of the late upstream region of HPV16 in cervical intraepithelial neoplasm and invasive carcinoma. AB - HPV16 is the most common type of human papillomavirus (HPV) seen in cervical squamous cell carcinoma. A 78-bp promotor element at nt 4118-4196 called late upstream region (LUR), critical for the expression of late genes, has been identified recently. Late genes encode viral capsid proteins that coat viral DNA to form particles and serve as antigen. To elucidate whether there are any sequence variations within LUR of HPV16 and any difference of these sequence variations between cervical invasive squamous cell carcinoma (CIC) and cervical intraepithelial neoplasia (CIN), we sequenced HPV16 LUR from 50 cases of HPV16 positive CIC and CIN. We found that variation frequency in the late upstream region ranged from 0 to 4.2 except for two cases in which variation frequency was as high as 22.8%. Eight of 24 CINs and 17 of 26 CICs contained two or more variations (33% vs. 65%, P < 0.025). The results suggested that the sequence variations occurred more often in LUR of HPV16 than in other regions of HPV16 and the variations in HPV16 LUR might play a role in the process of carcinogenesis of cervical carcinoma. PMID- 11860544 TI - Laminin-5 gamma 2 chain as an invasivity marker for uni- and multifocal lesions in the lower anogenital tract. AB - During recent decades it has become apparent that there are two types of vulvar disease: the classic type found in elderly women with unicentric and unifocal lesions, and the type found in younger women, in which precancerous and invasive changes develop in the anogenital lower tract in a multicentric and multifocal fashion, often over a long period of observation. The laminin-5 gamma 2 chain is an extracellular protein that is a component of the basement membrane. Recently its expression has been recognized as a marker in cervical cancer that permits identification of invasive capacity. The aim of our study was to determine if laminin-5 gamma 2 chain antibody can act as a sensitivity marker of invasive capacity in precancerous and invasive carcinoma in women with uni- and multifocal changes in the anogenital tract. The result showed that all patients in the older group of women with invasive carcinoma of the vulva had moderate to high positive expression of the laminin-5 gamma 2 chain. In the group of younger patients with multifocal precancerous changes observed over long periods, most of the patients with vulva intraepithelial neoplasia (VIN) 3 showed laminin-5 gamma 2 chain positivity already in the precancerous changes, and all of them developed invasivity during the period of observation. Normal epithelium without atypia was mostly negative or of low immunoreactivity of laminin-5. In conclusion, positive laminin-5 gamma 2 chain expression seems to indicate the invasiveness potential of precancerous lesions and is also expressed in all investigated invasive carcinomas of the anogenital tract. PMID- 11860545 TI - Nuclear DNA content, proliferative activity, and p53 expression related to clinical and histopathologic features in endometrial carcinoma. AB - The purpose of this study was to evaluate the prognostic impact of image cytometry DNA ploidy, MIB-1, and p53 in relation to clinicopathologic variables in 376 consecutive patients with endometrial carcinoma stages I-IV. Following primary treatment 358 patients were considered tumor-free. Relapses and tumor specific deaths of these patients were noted. Image cytometry DNA ploidy (n = 340) and expression of MIB-1 (n = 318) and p53 (n = 323) were studied. In univariate analysis, stage (P < 0.001), histopathologic subtype (P < 0.001), degree of differentiation (P < 0.001), HRT (P = 0.034), DNA ploidy (P < 0.001), and p53 (P < 0.001) were significant predictors of relapse. Patient age showed that the estimated mean risk of relapse increases with nearly 64% per decade in life (P 0.003), and the MIB-1 expression with 21% per 10-unit increment (P 0.004). In multivariate analysis, degree of differentiation, MIB-1, and p53 lost their prognostic capability. However, after stage and histopathologic subtype, image cytometry DNA ploidy was the strongest predictor of outcome and was of value in predicting the risk for relapse. The combination of DNA ploidy, MIB-1, and p53 expression was an even stronger predictor of relapse-free survival than the individual prognostic factors. PMID- 11860546 TI - Postoperative chemotherapy in advanced ovarian granulosa cell tumors. AB - The objective of this research is to assess the use of first-line postoperative chemotherapy in patients with advanced ovarian granulosa cell tumor (GCT). A retrospective population-based case series identified 60 women with stage IC or greater ovarian GCT over a 25-year period. Five patients were excluded because of incomplete information. None of the patients had received chemotherapy or radiotherapy prior to the diagnosis of advanced GCT. All patients had, at a minimum, a total abdominal hysterectomy and bilateral salpingo-oophorectomy. Pathology was centrally reviewed and the diagnosis confirmed. Of the 55 eligible patients, the 21 women with stage III and IV disease were the main focus of the study. Clinical outcomes and survival were compared between 13 women who received combination chemotherapy and eight who did not. Univariate analysis was conducted to assess the impact of age at diagnosis, size of residual disease, and adjuvant use of radiation therapy on prognosis. For the 55 patients, median age at diagnosis was 54 years (range 22-79). Median length of follow-up was 4.4 years (range 0.3-23.3). Median time to progression was 2.3 years (range 0.3-5.3). Sixty percent of those with no macroscopic disease after primary surgery recurred within 4.5 years of diagnosis. All patients with gross residual disease (>2 cm) were dead within 4 years of diagnosis. Overall 5 years survival rate was 61.6% (95% CI (49.3-76.9)). Among stage III and IV patients, there were no differences with respect to age at diagnosis and use of radiation therapy between those who did and did not receive chemotherapy. The only statistically significant difference was the presence of macroscopic residual disease (82% vs. 22%). Although there was no statistical significant difference in overall survival, there was a trend toward a poorer outcome in the group that received chemotherapy. Survival of patients with macroscopic residual disease was not influenced by use of chemotherapy (P = 0.976). We conclude that the presence of macroscopic residual disease after primary surgery was the most important prognostic factor. Although these patients were more likely to receive postoperative chemotherapy, there was no evidence to document a beneficial effect of systemic therapy in this group of women. PMID- 11860547 TI - Clear cell sarcoma arising in the retroperitoneum. AB - Clear cell sarcoma is a rare soft-tissue neoplasm, arising most commonly in the tendons and aponeuroses of young adults. We report here the first female case of clear cell sarcoma arising in the retroperitoneum with clinical features similar to those of malignant ovarian tumors. Aspects of clinical presentation, histopathologic evaluation, and treatment are described. PMID- 11860548 TI - Rhabdomyosarcoma of the uterus: report of two cases, including one of the spindle cell variant. AB - Most uterine sarcomas fall into the category of leiomyosarcoma, endometrial stromal sarcoma, or undifferentiated sarcoma. Pure rhabdomyosarcomas are extremely rare, although a rhabdomyosarcomatous element may be present as a component of an adenosarcoma or carcinosarcoma (malignant mixed mullerian tumor). This report describes two uterine rhabdomyosarcomas in 28- and 67-year-old women. These were of spindle cell and pleomorphic types, respectively. At presentation the pleomorphic rhabdomyosaroma was stage IV, exhibiting massive pelvic and abdominal dissemination that mimicked an ovarian neoplasm. The spindle cell rhabdomyosarcoma was stage I, being confined to the uterus. Grossly, both uterine tumors had a polypoid appearance. Immunohistochemically, tumor cells were positive with the skeletal muscle markers sarcomeric actin, myoglobin, and myoD1. The patient with stage IV disease died within a short time of diagnosis and the other patient is alive and well at 2 years' follow-up. This report adds to the published literature on uterine rhabdomyosarcomas. This is the first reported uterine case of the spindle cell variant of embryonal rhabdomyosarcoma. Based on these cases and the published literature, rhabdomyosarcoma, especially the pleomorphic variant, appears to be a very aggressive neoplasm with an extremely poor prognosis. Immunohistochemical demonstration of skeletal muscle differentiation is necessary for a definitive diagnosis. PMID- 11860549 TI - The transcriptional regulation of the Streptococcus mutans bgl regulon. AB - A beta-glucoside utilization regulon recently isolated from Streptococcus mutans has been shown to contain genes involved in beta-glucoside hydrolysis and a putative regulator. The bglP gene encodes a beta-glucoside-specific enzyme II (EII) component of the phosphoenolpyruvate-dependent phosphotransferase system, the bglC gene encodes a putative transcriptional regulator, and the bglA gene encodes a putative phospho-beta-glucosidase. To investigate the transcriptional activity of these genes, the putative promoter regions of the bglP, bglC and bglA genes were fused with the E. coli lacZ reporter gene. The resultant reporter plasmids were used to monitor the transcriptional activity of these loci in S. mutans. The results illustrate that these genes are not repressed by glucose in the presence of an inducing beta-glucoside, esculin, to the levels of expression observed in the absence of esculin. Therefore, these loci are not subject to catabolite repression by glucose to noninduced levels of expression. The bglC gene product was determined to be a positive transcriptional regulator of the bglA gene but does not regulate the expression of the bglP gene. Thus, regulation of these loci requires different and multiple control mechanisms. PMID- 11860550 TI - Stability and activity of specific antibodies against Streptococcus mutans and Streptococcus sobrinus in bovine milk fermented with Lactobacillus rhamnosus strain GG or treated at ultra-high temperature. AB - Passive local immunization against dental caries is a promising approach to its prevention, as clinical evidence of active oral or nasal immunization is still limited and controversial. By means of systemic immunization of pregnant cows with a multivalent vaccine, high titres of IgG antibodies against human cariogenic bacteria, Streptococcus mutans and Streptococcus sobrinus, were produced in bovine colostrum. The purified immune product (IP) of this preparation has a number of anticariogenic properties, such as inhibition of streptococcal adherence to saliva-coated hydroxyapatite and inhibition of glucosyltransferase enzymes. This study investigated whether IP antibodies remained active and functional when added to ultra-high temperature (UHT)-treated milk or to Lactobacillus rhamnosus GG (LGG)-fermented milk stored for an extended time. LGG was chosen because of its widely known health benefits in humans and animals. A commercial UHT toddler's milk was supplemented with IP and stored for 2 months at 5, 21 and 30 degrees C. The antistreptococcal titres in UHT milk did not decline at any temperature during storage, and UHT-IP inhibited the adherence of S. mutans for up to 2 months. This was not the case with UHT toddler's milk without IgG antibodies. Milk was fermented with live LGG cells in the presence or absence of 5% IP. The antistreptococcal titres declined to about 30% of the original titres after storage. Fresh milk alone slightly enhanced streptococcal adhesion but fresh milk with IP inhibited the adherence of S. mutans by over 50%. LGG-positive fermented milk without antibodies also inhibited (P < 0.05) the adhesion by about 40%. In both LGG-fermented and UHT immune milk, the activity of antibodies against cariogenic streptococci was maintained during the expected shelf-life of these products. From the anticariogenic point of view it may be beneficial to add bovine-specific antibodies against mutans streptococci to probiotic LGG-containing milk products. PMID- 11860551 TI - Antigenic properties of the GroEL-like protein of Campylobacter rectus. AB - The purpose of this study was to clarify the antigenic properties of the GroEL like protein of Campylobacter rectus using a specific polyclonal antibody directed to the purified 64-kDa GroEL-like protein (pAb-CrGroEL), a polyclonal antibody directed to the Actinobacillus actinomycetemcomitans GroEL-like protein (pAb-AaGroEL) and a monoclonal antibody against the recombinant human HSP60 (mAb HuHSP60). In SDS-PAGE/Western immunoblotting analysis, mAb-HuHSP60, pAb-CrGroEL and pAb-AaGroEL were found to react with the GroEL-like protein (64-kDa) present in all C. rectus strains. A 150-kDa protein in C. rectus ATCC 33238 also reacted strongly with pAb-CrGroEL. This 150-kDa protein was found to be present on the surface-associated material of bacterial cells, as determined by transmission electron microscopy and immunogold labelling of cells with pAb-CrGroEL. Analysis of the first 20 N-terminal amino acids of the sequence of the 150-kDa protein revealed a strong homology (80%) with the C. rectus surface layer (S-layer) protein. Investigation of the biochemical nature of antigenic determinants using periodic acid and proteolytic enzymes showed that the C. rectus GroEL-like protein possessed immunodominant epitopes in both peptide and carbohydrate chains, and that the immunoreactive determinants of the 150-kDa protein belonged to carbohydrate. These results suggest that the GroEL-like protein and the S layer protein of C. rectus may share the same carbohydrate epitopes. PMID- 11860552 TI - Initial characterization of the Streptococcus gordonii htpX gene. AB - Examination of the Streptococcus gordonii chromosomal region, which lies immediately upstream of the glucosyltransferase positive regulatory determinant rgg, revealed two open reading frames. Based on nucleotide sequences, these genes were similar to the Listeria monocytogenes lemA gene, which is involved in antigen presentation, and the Escherichia coli htpX heat shock gene, which has an unknown function. Northern hybridization analysis indicated that S. gordonii lemA and htpX genes were associated with a ca. 1.7-kb polycistronic transcript. Although levels of the lemA/htpX transcript did not increase in response to heat to levels seen with dnaK controls, insertional inactivation of htpX resulted in changes in adhesiveness, cellular morphology and detergent-extractable surface antigens in cells grown at 41 degrees C, implying that htpX may be involved in surface protein expression. Insertional inactivation of lemA and htpX indicated that, despite their proximity to rgg and the structural gene, gtfG, these upstream genes do not affect S. gordonii glucosyltransferase activity. PMID- 11860553 TI - The effect of tissue molecules on bacterial invasion of dentine. AB - Bacterial invasion of dentinal tubules is a critical step in the pathogenesis of dental caries and pulp and periapical disease. The purpose of this study was to determine the effect some molecules commonly found in saliva and dentinal tubule fluid may have on the bacterial invasion of dentine. The results showed that invasion of Streptococcus gordonii or Enterococcus faecalis cells was inhibited when the bacterial cells were in solution with mucin, immunoglobulin G (IgG) and serum, and this was related to bacterial cell aggregation, as a result of interaction with agglutinins, and/or inhibition of collagen binding. When dentine was soaked in growth media containing fibrinogen, IgG, albumin or serum prior to inoculation, bacterial invasion was inhibited. It is suggested that this may be due to reduced dentine permeability as a consequence of the deposition of the compounds within dentinal tubules. PMID- 11860554 TI - Th1/Th2 cytokine profiles in saliva of HIV-positive smokers with oropharyngeal candidiasis. AB - Oropharyngeal candidiasis (OPC) is a common opportunistic infection among HIV positive individuals and often correlates with a CD4 cell number < 200 cells/microl. This study further examined the association of smoking and OPC in HIV-positive persons. A strong association between smoking and OPC was seen in HIV-positive individuals with > or =200 CD4 cells/microl. In HIV-positive persons with > or =200 CD4 cells/microl, OPC+ smokers had lower gamma-interferon (IFN gamma) concentrations and a trend toward higher interleukin (IL)-4 concentrations in whole saliva compared to OPC- persons with > or =200 CD4 cells/microl, a cytokine profile consistent with that observed in HIV+OPC+ persons with < 200 CD4 cells/microl. These results suggest that premature OPC in HIV-positive smokers is associated with altered oral host defence mechanisms that cannot be overcome by levels of systemic CD4 cells that are otherwise sufficient to protect against OPC. PMID- 11860555 TI - Molecular characterization of Candida spp. isolated from the oral cavities of patients from diverse clinical settings. AB - Infections by Candida spp. have increased in medical importance over the past few decades. Our understanding of species identification, commensalisms, pathogenicity, person-to-person spread, and the development of antifungal resistance within specific strains has been greatly enhanced by the utilization of molecular epidemiological methodology. The aim of the current research was to assess the quantity, species and molecular characterization of oral yeast isolates from well-defined cohorts of immunocompetent patients from a diverse range of clinical settings. Oral rinse samples were assessed for the growth of yeast and degree of colonization. Isolates were defined to the species level by both phenotypic and molecular methods and strains were further genotypically subtyped. Significant variation was shown to exist in the number, species and genotypic subgroups of yeast isolated from the oral cavity in different patient groups. This variation could be attributed to the local oral conditions unique to these patient groups. PMID- 11860556 TI - Dipeptide utilization by the periodontal pathogens Porphyromonas gingivalis, Prevotella intermedia, Prevotella nigrescens and Fusobacterium nucleatum. AB - Porphyromonas gingivalis, Prevotella intermedia, Prevotella nigrescens and Fusobacterium nucleatum, which can frequently be isolated from periodontal pockets, preferentially utilize proteins and peptides as growth substrates. In this study, we determined the size of peptide that is preferentially utilized as a source of energy and material for cell growth by P. gingivalis, P. intermedia, P. nigrescens and F. nucleatum using various sizes of poly amino acids consisting of two to approximately 100 molecules of aspartate or glutamate. Resting cells of P. gingivalis, P. intermedia and P. nigrescens utilized aspartylaspartate, while cells of P. gingivalis and F. nucleatum utilized glutamylglutamate. The addition of aspartylaspartate to the culture medium increased the growth of P. gingivalis, P. intermedia and P. nigrescens, while the addition of glutamylglutamate promoted the growth of P. gingivalis and F. nucleatum. These results clearly indicate that dipeptides such as aspartylaspartate and glutamylglutamate can be utilized as growth substrates for P. gingivalis, P. intermedia, P. nigrescens and F. nucleatum. PMID- 11860557 TI - Periodontal pathogen detection in gingiva/tooth and tongue flora samples from 18- to 48-month-old children and periodontal status of their mothers. AB - Few studies have detected periodontal pathogens in young children, and when detected the prevalence has been relatively low. In this epidemiological study, we determined the prevalence of periodontal pathogen colonization in young children and examined the relationship between periodontitis in mothers and detection of periodontal pathogens in their children aged 18-48 months. Children were selected and enrolled randomly into the study; tongue and gingival/tooth plaque samples were harvested and analyzed by DNA probe checkerboard assay for Porphyromonas gingivalis and Bacteroides forsythus. Clinical measurements included a gingival bleeding score in the children and a periodontal screening and recording (PSR) score in the mothers. Mothers having one or more periodontal sites with probing depths > 5.5 mm were classified as having periodontitis. In this population, 71% (66/93) of the 18- to 48-month-old children were infected with at least one periodontal pathogen. Detection rates for children were 68.8% for P. gingivalis and 29.0% for B. forsythus. About 13.8% (11/80) of children had gingival bleeding in response to a toothpick inserted interproximally. Children in whom B. forsythus was detected were about 6 times more likely to have gingival bleeding than other children. There was no relationship between bleeding and detection of P. gingivalis. 17.0% (16/94) of the mothers had periodontitis. When all mother-child pairs were considered, the periodontal status of the mother was found not to be a determinant for detection of periodontal pathogens in the floral samples from the children. However, the odds ratio that a daughter of a mother with periodontitis would be colonized was 5.2 for B. forsythus. A much higher proportion of children in this population were colonized by P. gingivalis and/or B. forsythus than has been previously reported for other populations. A modest level of association between manifestations of periodontitis in mothers and detection of B. forsythus in their daughters was observed. PMID- 11860558 TI - Candida attachment to oral epithelium. AB - Inflamed oral mucosa biopsies from patients with thrush and high candidal density were observed in a transmission electron microscope (TEM) using ultra histochemical staining with ruthenium red for glycocalyx visualization. Fimbriae comprising the glycocalyx and enabling yeast adhesion to epithelial cells were clearly visualized by ruthenium red. All internalized portions of the yeast walls were devoid of glycocalyx, indicating that the growing tips of the hyphae mechanically penetrated the host cells. The attachment of Candida occurred in two ways: by fimbria-mediated adhesion enabling colonization of the epithelial surface, and by invasion of the superficial epithelial cells via hyphae. As the interaction between adhesin receptors and adhesins stimulates the production of proinflammatory cytokines, Candida adhesion itself is assumed to induce mucosal inflammation. PMID- 11860561 TI - Sonographic findings in a series of rapidly involuting congenital hemangiomas (RICH). AB - We report 10 patients with lesions consistent with the clinical entity of rapidly involuting congenital hemangioma (RICH). These are congenital vascular tumors bearing some resemblance to infantile hemangiomas, but with important clinical differences. The lesions in our patients were firm, red or purple plaques or tumors, sometimes with surface telangiectasia and almost all with a pale or blanched halo. They were fully developed at birth and underwent no further expansion. All promptly began to resolve and this progressed rapidly over the early months of life, with complete resolution, sometimes with residual atrophy, occurring at less than 1 year of age in most of the patients. Sonography was performed in all of our cases. A remarkably consistent picture was demonstrated. The lesions were uniformly hypoechoic and mostly confined to the subcutaneous fat. They were diffusely vascular, being traversed by multiple tubular vascular channels. Some of the channels were compressible, with a venous flow signal, while others demonstrated low resistant arterial flow. These features are consistent with recently described histopathologic findings in these rapidly involuting lesions. We feel that this entity can be reliably diagnosed in most cases with a consideration of the unique clinical features and with the use of noninvasive imaging studies. PMID- 11860562 TI - Juvenile acanthosis nigricans and insulin resistance. AB - Acanthosis nigricans in obese adolescents is frequently associated with hyperinsulinemia and insulin resistance. We report three cases of this condition. In the early stage the skin lesions appeared to be erythematous and pruritic, mimicking an inflammatory dermatitis. Dietary control and oral metformin hydrochloride markedly improved the lesions. Topical calcipotriol may also help to control the skin condition. PMID- 11860563 TI - Infantile perianal protrusion: 13 new cases. AB - We report 13 girls, ages 4 months-5 years, affected by an infantile perianal protrusion located on the midline anterior to the anus in 8 patients and posteriorly in 3. A past personal history of constipation existed in six patients and anal fissures in three patients. In one of the five patients with recurrent anogenital erythema, a punch biopsy specimen revealed the histopathologic signs of lichen sclerosus et atrophicus (LS&A). In another two patients, infantile perianal protrusion was associated with clinical signs of genital LS&A. None of our patients had spontaneous reduction of infantile perianal protrusion during the follow-up period. PMID- 11860564 TI - Annular pustular psoriasis--most common form of pustular psoriasis in children: report of three cases and review of the literature. AB - Annular pustular psoriasis (APP) is a rare form of pustular psoriasis with a chronic recurrent course and good prognosis. We report three cases of APP in children, two of whom were siblings. Review of the medical literature reveals that a disproportionately high percentage of cases of APP occur in children. In some cases topical therapy can clear the condition, although in severe or recalcitrant disease, systemic therapy may be necessary. PMID- 11860565 TI - Eruptive vellus hair cysts presenting as multiple periorbital papules in a 13 year-old boy. AB - Eruptive vellus hair cysts (EVHCs) are small, cystic papules that usually occur on the chest and proximal extremities of children and adults. The etiology is unknown. EVHCs of the eyelids have been reported infrequently. We describe a 13 year-old boy who presented with symptomatic EVHCs in a periorbital distribution. PMID- 11860566 TI - Giant cell fibroblastoma in a child misdiagnosed as a dermatofibroma. AB - We report a 9-year-old African-American boy with a giant cell fibroblastoma of the shoulder that was incorrectly diagnosed as a keloid and dermatofibroma. Initial misdiagnosis led to a delay of 4 years in the correct diagnosis, with the tumor producing significant local destruction. We review herein the clinical manifestations, histologic findings, histogenesis, relationship to dermatofibrosarcoma protuberans (DFSP), treatment, and differential diagnosis of giant cell fibroblastoma (GCF). This information is important in correctly diagnosing this uncommon, benign, but locally aggressive and recurrent tumor of childhood. The clinician should consider GCF and DFSP when the pathologic diagnosis of dermatofibroma is made in lesions more than 2 cm in diameter, or when this diagnosis is made in a prepubertal child. PMID- 11860567 TI - Cutaneous focal mucinosis: a case report. AB - Cutaneous or dermal mucinoses are a heterogeneous group of connective tissue disorders in which there is an accumulation of mucin in the skin or within the hair follicle. They have a number of different morphologic presentations and can often be confused with other diseases. We report a 12-year-old Chinese girl with a 1-year history of a well-defined, hypopigmented plaque on her chin, consistent with cutaneous focal mucinosis, a rare condition in children. PMID- 11860568 TI - Traditions, anthrax, and children. AB - Anthrax is sporadically seen in Turkey, especially among people who live in rural areas and who come in contact with animals. Two siblings with cutaneous anthrax are described in this report. A week before their admission to the hospital, contaminated cow's blood was smeared on their foreheads as part of a traditional ritual. Both children were successfully treated with crystalline penicillin. In developing countries, traditions such as blood smearing may be an important factor in the transmission of anthrax to children. PMID- 11860569 TI - Pyogenic granuloma of the penile shaft following circumcision. AB - Pyogenic granuloma is a benign vascular, inflammatory proliferation that appears following minor trauma in children. The exact pathogenesis of pyogenic granuloma is unknown, but its formation is closely related to minor trauma, chronic irritation, and hormonal influences. We present a unique case of a pyogenic granuloma that appeared in an infant following circumcision. PMID- 11860570 TI - Ulerythema ophryogenes in Cornelia de Lange syndrome. AB - A 17-year-old woman with Cornelia de Lange syndrome had asymptomatic skin lesions since the age of 4 years. These were multiple, follicular, horny papules, present on both cheeks, and surrounded by erythematous skin. Similar lesions were present on the external aspect of the arms, but amidst skin of normal coloration. Keratosis pilaris atrophicans faciei (ulerythema ophryogenes) has been described as a cutaneous marker for several congenital syndromes. To our knowledge, its possible association with Cornelia de Lange syndrome has never been reported. PMID- 11860571 TI - Atypical pityriasis rubra pilaris associated with arthropathy and osteoporosis: a case report with 15-year follow-up. AB - Pityriasis rubra pilaris (PRP) is an idiopathic papulosquamous disease that clinically presents with palmoplantar keratoderma and follicular hyperkeratotic papules that coalesce into scaly erythematous plaques. We report a unique case of atypical PRP beginning at 1 year of age with associated severe arthropathy and osteoporosis. We further discuss the clinical and histopathologic aspects of PRP, its possible etiology, and other associated conditions. PMID- 11860572 TI - Stevens-Johnson syndrome developing in a girl with systemic lupus erythematosus on high-dose corticosteroid therapy. AB - Stevens-Johnson syndrome (SJS) is a vesiculobullous disease of the skin and mucosa. This generalized hypersensitivity reaction is well known to occur in association with certain drugs, particularly sulfonamides, nonsteroidal anti inflammatory agents (NSAIDs), and anticonvulsants. The disease is usually seen in children and young adults and is often treated with corticosteroids. We present a 9-year-old girl with lupus nephritis who developed cutaneous bullae and mucositis while being treated with intravenous methylprednisolone. The initial differential diagnosis included bullous lupus, but skin biopsy specimen findings supported a diagnosis of SJS. She was treated with intravenous immunoglobulin (IVIg). PMID- 11860573 TI - Pityriasis lichenoides in a girl with the granulomatous form of common variable immunodeficiency. AB - Pityriasis lichenoides (PL) is a cutaneous disease of unknown origin. In our 8 year-old female patient with the granulomatous form of common variable immunodeficiency (CVID), PL occurred together with massive splenomegaly and intra abdominal lymphadenopathy. Prednisone was efficient for treatment of her splenomegaly and autoimmune cytopenias. However, PL was resistant to both topical and systemic steroid treatment. Healing of PL was achieved with the use of a super-potent topical steroid, clobetasol propionate. A defect of T-cell function in CVID may contribute to development of PL. In the granulomatous form of CVID, sarcoid-like granulomas are the most commonly reported cutaneous lesions. PL has not been previously reported. PMID- 11860574 TI - Relapsing polychondritis: a pediatric case. AB - A pediatric case of relapsing polychondritis is reported. Diagnostic features were recurrent episodes of painful inflammation of ear cartilage, episcleritis, and atopic disease. A good therapeutic response was obtained with systemic corticosteroids, dapsone, and methotrexate. PMID- 11860575 TI - Giant nevoid hypertrichosis in an Iranian girl. AB - A 12-year-old Iranian girl and her father had an extensive congenital nevus consisting of long, black terminal hair involving the entire back and the posterior surface of the arms without pigmentary changes of the underlying skin. A physical examination revealed no other abnormalities. In this study the possibility of a genetically determined etiology is considered. PMID- 11860576 TI - Restrictive dermopathy: a case report and a critical review of all hypotheses of its origin. AB - Restrictive dermopathy (RD) is a rare, fatal, autosomal recessive genodermatosis in which tautness of a translucent thin skin is the major clinical observation. This causes an intrauterine fetal akinesia deformation sequence (FADS) resulting in polyhydramnios, reduced fetal movements at around 31 weeks gestation, dysmorphic facies, arthrogryposis, and early neonatal death because of respiratory insufficiency. The characteristic histologic abnormalities of the skin are located in a thin dermis, consisting of compactly arranged collagen fibers, scant elastic fibers, and poorly developed skin appendages. The epidermal rete ridges are flattened and the dermal-hypodermal border is remarkably straight. The etiology of these changes remains unclear. We tested several existing hypotheses and could not confirm them. These included fibroblast dysfunction, abnormal keratin composition, desmosomal changes, and increased proinflammatory cytokines [tumor necrosis factor (TNF)-alpha and interleukin (IL) 6]. We conclude that RD is a relatively easy clinical and pathologic diagnosis, but that the pathogenesis of the disease is not clarified. PMID- 11860577 TI - Acquired leukoderma in congenital pigmented nevus associated with vitiligo-like depigmentation. AB - We report a 6-year-old boy who developed depigmentation within a congenital melanocytic nevus at the age of 3 years. During the following months a halo phenomenon and vitiligo-like lesions distant from the nevus appeared. A thorough search for malignant melanoma was negative. A second patient, a 45-year-old woman, had a large congenital nevus on the trunk with marked satellitosis. At the age of 20 years, partial regression of the large nevus occurred and, in addition, halos developed around almost all smaller nevi. Repeated searches for associated malignant melanoma were negative. We review the rare cases of halo congenital nevi and emphasize that depigmentation is not necessarily associated with malignant degeneration. PMID- 11860578 TI - Eruptive pseudoangiomatosis. AB - We describe two children with acute onset and spontaneous resolution of angioma like papules during a viral illness. The biopsy specimens from both patients showed a unique histologic appearance consisting of dilated dermal blood vessels with plump, hobnail-shaped endothelial cells. On the basis of the natural history and the histopathologic features we suggest the diagnosis of eruptive pseudoangiomatosis. PMID- 11860579 TI - Clotrimazole/betamethasone diproprionate: a review of costs and complications in the treatment of common cutaneous fungal infections. AB - The use of antifungal/corticosteroid combinations as topical therapy for dermatophytoses has been criticized as being less effective, more expensive, and the cause of more adverse cutaneous reactions than antifungal monotherapy. The combination of clotrimazole and betamethasone diproprionate (Lotrisone) is a mix of an azole antifungal and a high-potency corticosteroid, and is one of the most widely prescribed of these combinations. Our objective was to describe the beneficial and deleterious effects of Lotrisone in the treatment of common cutaneous fungal infections and its relative cost-effectiveness. We did a literature review documenting clinical trial data and adverse reactions to Lotrisone and collected a cost analysis of topical antifungal prescribing data over a 2-month period from a large midwestern staff-model health maintenance organization (HMO). Lotrisone is approved by the U.S. Food and Drug Administration (FDA) for the treatment of tinea pedis, tinea cruris, and tinea corporis in adults and children more than 12 years of age. Treatment is limited to 2 weeks in the groin area and 4 weeks on the feet. The most concerning adverse effects of Lotrisone were reported in children and included treatment failure, striae distensae, hirsuitism, and growth retardation. This combination was also reported to have decreased efficacy in clearing candidal and Trichophyton infections as compared to single-agent antifungals. Lotrisone was considerably more expensive than clotrimazole alone and was found to account for more than 50% of topical antifungal expenditures as prescribed by primary care physicians, but only 7% of topical antifungals prescribed by dermatologists. We found that Lotrisone was shown to have the potential to induce many steroid-related side effects and to be less cost effective than antifungal monotherapy. This combination should be used judiciously in the treatment of cutaneous fungal infections and may not be appropriate for use in children. PMID- 11860580 TI - What syndrome is this? Fabry disease in a heterozygous female. PMID- 11860583 TI - Perforation of the nasal septum and palatum durum in a severely malnourished infant. PMID- 11860584 TI - Multiple metastatic tuberculous abscesses in a severely malnourished infant. PMID- 11860585 TI - Pachyonychia congenita affecting only the nails. PMID- 11860587 TI - Pellagrous dermatitis induced by phenytoin. PMID- 11860586 TI - Acute infantile hemorrhagic edema and Henoch-Schonlein purpura overlap in a child. PMID- 11860588 TI - Keratomalacia and scurvy in a severely malnourished infant. PMID- 11860589 TI - A case of chickenpox associated with facial nerve palsy. PMID- 11860590 TI - Erythema multiforme: a practical approach to recent advances. PMID- 11860591 TI - Presenteeism...or when being there is not being there. PMID- 11860592 TI - A win-win model for an academic nursing center: community partnership faculty practice. AB - A number of schools of nursing have established community nursing centers to provide faculty practice sites, student learning experiences, and a service to the community, most often to a poor underserved population. The current literature concludes that these centers provide a quality clinical service and improve access to health care, and they also provide an avenue for research, training, and faculty practice. Acquiring necessary financial support and the ability to achieve financial independence appear to be the most common difficulties for these centers. Most of the current literature includes an examination of issues relating to funding. The model presented in this article focuses on organizational variables that include both the center and its placement in relationship to other functions and programs in the school and a broadening of the meaning of fiscal responsibility to include an awareness of the broad spectrum of benefits that the community nursing center brings to the entire school. Efforts to coordinate and integrate the needs and functions of several groups are described. Establishing goals and priorities that simultaneously meet the needs of all or most of these groups has been an important outcome. The activities of the center have become an integral part of the everyday life of the school. Achieving financial independence and being fiscally aware and responsible is not the same thing. PMID- 11860593 TI - Perception of risk by administrators and home health aides. AB - Increasing numbers of persons over 65, decreased length of hospital stay, and need for chronic (custodial) health care have placed a strain on home health care agencies. The second largest group of persons providing care is home health aides (HHAs), who perform in-home, nonskilled, technical procedures with little or no on-site supervision. They are generally high school graduates or hold GEDs. The purpose of this study was to compare home health care administrators' (HHCAs) and HHAs' perceptions of risk involved in home visiting. Given HHAs' educational preparation and limited supervision, they are basically on their own for work performed. Although agencies provide orientation sessions for new workers, periodic in-services often relate to tasks and competency testing and little attention directed toward protecting the self-specifically, strategies to decrease personal risk. In order to determine to what extent HHCAs and HHAs perceive risk, the Home Health Care Perception of Risk Questionnaire, a self report measure, was administered to a national random sample of 93 HHCAs and 227 HHAs. Findings suggest that these groups differ in perception of risk and level of agency support in making home visits. Suggestions for meeting the needs of this HHA provider group are offered. PMID- 11860594 TI - Home care nursing in Japan: a challenge for providing good care at home. AB - Home care in Japan has developed over the past 30 years. Nurses have taken leadership in promoting home care and at the same time have expanded their roles. The roles of Japanese nurses in the field of home care are presented in the context of the historical perspective and view for the future. Home care nurses have performed care management for their community as well as for individual older clients living in their community. Currently, nurses work as high-tech and personal care providers and are developing a new role in health care enterprises. The number of nurses working as clinical nurse specialists will increase consistently with the rapid increase of master's programs. In the future, nurses should take a lead in developing health policy. The purpose of the article is to describe the current situation of home care clients and home care nursing in Japan. In addition, four issues of home care in Japan are described related to home care for older persons, high-tech home care, terminal care, and home care for psychiatric patients. PMID- 11860595 TI - Maternal resources, parenting, and dietary patterns among rural African American children in single-parent families. AB - A correlation study was designed to test the hypothesis that maternal education, perceived family resources, and the importance of family routines would be related to children's dietary patterns. Additionally, the study examined the hypothesis that dietary patterns would be associated with children's cognitive and physical abilities. The sample for this study included 159 African American single-mother families with a 6- to 9-year-old child living in rural areas, most of whom lived in poverty. Children's eating patterns were assessed using a self report questionnaire administered to the mother in an interview format. Children's cognitive ability was measured by several subscales from the Woodcock Johnson Psycho-Educational Instrument and the Harter Perceived Competence Scale for children. For male children, the mother's higher education was related to more adequate eating patterns at home, and more perceived family resources were related to the likelihood of taking vitamin supplements. For female children, greater milk intake was positively related to cognitive outcomes, including applied problem, passage comprehension, calculation, synonym identification, antonym identification, and quantitative concept scores. Milk intake was clearly related to more optimal cognitive development. The results of this study support the literature related to the importance of nutrition for cognitive and physical abilities in children. PMID- 11860596 TI - Developing a school asthma policy. AB - Schools are faced with the challenging mandate of addressing the learning needs of students while simultaneously managing a gamut of behavior and health problems. School health policies have been successfully used for many health related issues. Although asthma is the most common chronic disease among children, schools receive only a small amount of support to manage asthma issues. This article describes our experiences in developing an asthma policy in schools. The problem of asthma in schools was assessed as part of a comprehensive community-based asthma intervention, and a plan was established to develop a school asthma policy. The goal of the policy was to facilitate a physical and social environment that enabled students with asthma to control optimally their condition, have a good quality of life, and learn effectively. Specific objectives included enhancing environmental control, educating staff, clarifying medication protocols, and responding appropriately to symptoms. Strategies in developing the policy included strengthening collaboration and networks among health, school, and community sectors; defining the role of the school and health sectors; designing the policy; refining the policy based on stakeholder feedback; and developing an evaluation plan. PMID- 11860597 TI - A community-based approach to sexual assault. AB - The community-based Athens-Clarke County Sexual Assault Nurse Examiners (ACC SANE) program was developed to respond to the needs of sexual assault victims. Although this type of SANE model allows for a strong nurse-managed program, there are frustrations and pitfalls that differ from most hospital-based programs. At this time, the majority (71%) of SANE programs are based in hospitals, and few (15%) function in outpatient/community facilities. In the ACC-SANE program, many community agencies interact to provide a seamless approach to postassault health care, counseling, and criminal prosecution. SANE nurses are well equipped to provide forensic examinations in a caring, out-of-hospital environment. The program is housed in a remodeled cottage that rests on property owned by the Athens-Clarke County government and that is adjacent to the local police department. ACC-SANE is governed by a nonprofit board of directors. This management model has allowed the original organizing groups to remain active in the development of the program. As these agencies continue in the governance of the program, they remain committed to the success of ACC-SANE. The impact of program growth and published program guidelines is examined. PMID- 11860598 TI - Client perceptions of parish nursing. AB - The purpose of this study was to examine parish nursing from a client's perspective. Parish nursing is a relatively new health delivery model that has rarely been investigated. In order to describe the client perception, an ethnographic approach was used. The convenience sample included clients from two congregations in a southeastern Appalachian area served by parish nurses. Face-to face client interviews were conducted, and the Spradley's ethnographic approach to data analysis of transcripts was used. Each interview was analyzed separately by the research group for patterns and meanings reflecting the emic perspective. Five themes of client perception of parish nursing emerged from the data: (1) being available, (2) integrating spirituality and health, (3) helping us help ourselves, (4) exploring parish nursing, and (5) evaluating parish nursing.Clients perceived having a parish nurse as positive and beneficial for individuals, the congregation, the church, and community. Parish nursing was viewed as a useful, meaningful, and effective health intervention and setting, and parish nurses were viewed as effective and meaningful health providers. Further exploration of the effectiveness of this nursing delivery model is warranted. PMID- 11860599 TI - Living arrangements and health status in later life: a review of recent literature. AB - The living arrangements of older persons play a key role in their use of formal and informal care, as well as in their health and well-being. Nurses engaged in primary care, discharge planning, and home care are strategically positioned to contribute to an optimal fit between older persons and their home environment. This article describes the demographic significance of late-life living arrangements and proposes a model for organizing the complex web of factors associated with household composition and late-life migration. The article then summarizes qualitative and quantitative evidence in support of the proposed model. Key areas for nursing research and strategies for applying available research are identified. PMID- 11860601 TI - Cloning and characterization of the rat homologues of the Inhibitor of Apoptosis protein 1, 2, and 3 genes. AB - BACKGROUND: Inhibitor of Apoptosis (IAP) proteins are key intrinsic regulators of apoptosis induced by a variety of triggers. We isolated the rat Inhibitor of Apoptosis genes 1, 2 and 3 and characterized their tissue distribution and expression. RESULTS: Rat iap-1 encodes a protein of 67.1 kDa with 73 % and 89.2 % homology to human and mouse iap-1 respectively. Rat iap-2 encodes a protein of 66.7 kDa with 81.6 % and 89.3 % homology to human and mouse iap-2 respectively. Rat iap-3 encodes a protein of 56.1 kDa with 89.5 % and 93.1 % homology to human and mouse iap-3 respectively. We have generated rabbit polyclonal antibodies against all three rat IAP genes. Northern and Western blot analysis detected rat IAP transcripts and proteins in majority of the tissues examined. In addition, a shorter, alternatively spliced transcript corresponding to iap-2 was found in testes. CONCLUSIONS: We have identified three rat homologues of the IAP genes. The elevated expression of rat iap-1 and iap2 in testes suggests that these two genes play an important antiapoptotic role in spermatogenesis. PMID- 11860600 TI - Quantitation of the distribution and flux of myosin-II during cytokinesis. AB - BACKGROUND: During cytokinesis, the cell's equator contracts against the cell's global stiffness. Identifying the biochemical basis for these mechanical parameters is essential for understanding how cells divide. To achieve this goal, the distribution and flux of the cell division machinery must be quantified. Here we report the first quantitative analysis of the distribution and flux of myosin II, an essential element of the contractile ring. RESULTS: The fluxes of myosin II in the furrow cortex, the polar cortex, and the cytoplasm were examined using ratio imaging of GFP fusion proteins expressed in Dictyostelium. The peak concentration of GFP-myosin-II in the furrow cortex is 1.8-fold higher than in the polar cortex and 2.0-fold higher than in the cytoplasm. The myosin-II in the furrow cortex, however, represents only 10% of the total cellular myosin-II. An estimate of the minimal amount of this motor needed to produce the required force for cell cleavage fits well with this 10% value. The cell may, therefore, regulate the amount of myosin-II sent to the furrow cortex in accordance with the amount needed there. Quantitation of the distribution and flux of a mutant myosin II that is defective in phosphorylation-dependent thick filament disassembly confirms that heavy chain phosphorylation regulates normal recruitment to the furrow cortex. CONCLUSION: The analysis indicates that myosin-II flux through the cleavage furrow cortex is regulated by thick filament phosphorylation. Further, the amount of myosin-II observed in the furrow cortex is in close agreement with the amount predicted to be required from a simple theoretical analysis. PMID- 11860602 TI - Killing of Mycobacterium avium subspecies paratuberculosis within macrophages. AB - BACKGROUND: Mycobacterium avium subspecies paratuberculosis (M. paratuberculosis) is a facultative intracellular pathogen that resides within host macrophages during infection of ruminant animals. We examined survival of M. paratuberculosis infections within cultured macrophages to better understand the interplay between bacterium and host. RESULTS: Serial plating of M. paratuberculosis infected macrophage lysates on Herold's egg yolk medium showed that mycobacterial replication takes place between 0 and 24 hours post-infection. This initial growth phase was followed by a steady decline in viability over the next six days. Antibodies against M. paratuberculosis were affinity purified and used in conjunction with transmission electron microscopy to track the development of intracellular bacilli. Immunogold labeling of infected macrophages with antibody against M. paratuberculosis showed degraded intracellular mycobacteria that were unrecognizable by morphology alone. Conversely, when macrophages were heavily infected with M. paratuberculosis, no degraded forms were observed and macrophages were killed. CONCLUSIONS: We present a general description of M. paratuberculosis survival within cultured macrophages using transmission electron microscopy and viability counts. The results of this study provides further insight surrounding M. paratuberculosis-macrophage infections and have implications in the pathogenesis of M. paratuberculosis, a pathogen known to persist inside cattle for many years. PMID- 11860603 TI - Quality control and data-handling in multicentre studies: the case of the Multicentre Project for Tuberculosis Research. AB - BACKGROUND: The Multicentre Project for Tuberculosis Research (MPTR) was a clinical-epidemiological study on tuberculosis carried out in Spain from 1996 to 1998. In total, 96 centres scattered all over the country participated in the project, 19935 "possible cases" of tuberculosis were examined and 10053 finally included. Data-handling and quality control procedures implemented in the MPTR are described. METHODS: The study was divided in three phases: 1) preliminary phase, 2) field work 3) final phase. Quality control procedures during the three phases are described. RESULTS: Preliminary phase: a) organisation of the research team; b) design of epidemiological tools; training of researchers. Field work: a) data collection; b) data computerisation; c) data transmission; d) data cleaning; e) quality control audits; f) confidentiality. Final phase: a) final data cleaning; b) final analysis. CONCLUSION: The undertaking of a multicentre project implies the need to work with a heterogeneous research team and yet at the same time attain a common goal by following a homogeneous methodology. This demands an additional effort on quality control. PMID- 11860604 TI - Simpson's paradox and calculation of number needed to treat from meta-analysis. AB - BACKGROUND: Calculation of numbers needed to treat (NNT) is more complex from meta-analysis than from single trials. Treating the data as if it all came from one trial may lead to misleading results when the trial arms are imbalanced. DISCUSSION: An example is shown from a published Cochrane review in which the benefit of nursing intervention for smoking cessation is shown by formal meta analysis of the individual trial results. However if these patients were added together as if they all came from one trial the direction of the effect appears to be reversed (due to Simpson's paradox). Whilst NNT from meta-analysis can be calculated from pooled Risk Differences, this is unlikely to be a stable method unless the event rates in the control groups are very similar. Since in practice event rates vary considerably, the use a relative measure, such as Odds Ratio or Relative Risk is advocated. These can be applied to different levels of baseline risk to generate a risk specific NNT for the treatment. SUMMARY: The method used to calculate NNT from meta-analysis should be clearly stated, and adding the patients from separate trials as if they all came from one trial should be avoided. PMID- 11860605 TI - Pooling data for number needed to treat: no problems for apples. AB - OBJECTIVE: To consider the problem of the calculation of number needed to treat (NNT) derived from risk difference, odds ratio, and raw pooled events shown to give different results using data from a review of nursing interventions for smoking cessation. DISCUSSION: A review of nursing interventions for smoking cessation from the Cochrane Library provided different values for NNT depending on how NNTs were calculated. The Cochrane review was evaluated for clinical heterogeneity using L'Abbe plot and subsequent analysis by secondary and primary care settings.Three studies in primary care had low (4%) baseline quit rates, and nursing interventions were without effect. Seven trials in hospital settings with patients after cardiac surgery, or heart attack, or even with cancer, had high baseline quit rates (25%). Nursing intervention to stop smoking in the hospital setting was effective, with an NNT of 14 (95% confidence interval 9 to 26). The assumptions involved in using risk difference and odds ratio scales for calculating NNTs are discussed. SUMMARY: Clinical common sense and concentration on raw data helps to detect clinical heterogeneity. Once robust statistical tests have told us that an intervention works, we then need to know how well it works. The number needed to treat or harm is just one way of showing that, and when used sensibly can be a useful tool. PMID- 11860606 TI - Meta-analysis, Simpson's paradox, and the number needed to treat. AB - BACKGROUND: There is debate concerning methods for calculating numbers needed to treat (NNT) from results of systematic reviews. METHODS: We investigate the susceptibility to bias for alternative methods for calculating NNTs through illustrative examples and mathematical theory. RESULTS: Two competing methods have been recommended: one method involves calculating the NNT from meta analytical estimates, the other by treating the data as if it all arose from a single trial. The 'treat-as-one-trial' method was found to be susceptible to bias when there were imbalances between groups within one or more trials in the meta analysis (Simpson's paradox). Calculation of NNTs from meta-analytical estimates is not prone to the same bias. The method of calculating the NNT from a meta analysis depends on the treatment effect used. When relative measures of treatment effect are used the estimates of NNTs can be tailored to the level of baseline risk. CONCLUSIONS: The treat-as-one-trial method of calculating numbers needed to treat should not be used as it is prone to bias. Analysts should always report the method they use to compute estimates to enable readers to judge whether it is appropriate. PMID- 11860607 TI - A survey of physicians knowledge regarding awareness of maternal alcohol use and the diagnosis of FAS. AB - BACKGROUND: Alcohol is the most widely used drug in the world that is a human teratogen whose use among women of childbearing age has been steadily increasing. It is also probable that Fetal Alcohol Syndrome is under diagnosed by physicians. The objectives of this study were twofold: 1) to evaluate the experience, knowledge and confidence of family physicians with respect to the diagnosis of FAS and 2) to evaluate physicians awareness of maternal drinking patterns. METHODS AND PARTICIPANTS: A multiple choice anonymous questionnaire was sent to a randomly selected group of family physicians in the Metropolitan Toronto area. RESULTS: There was a 73% (75/103) total response rate; Overall, 6/75 (8%) of family physicians reported that they had actually diagnosed a child with FAS. 17.9% had suspicions but did not make a diagnosis and 12.7% reported making a referral to confirm the diagnosis. Physician rated confidence in the ability to diagnosis FAS was low, with 49% feeling they had very little confidence. 75% reported counselling pregnant women and 60.8% reported counselling childbearing women in general on the use of alcohol. When asked what screening test they used to detect the use of alcohol, 75% described frequency/quantity. Not a single respondent identified using the current accepted screening method for alcohol use (TWEAK) which is recommended by The Centre for Addiction and Mental Health. CONCLUSIONS: Family physicians do not feel confident about diagnosing FAS. None of the physicians were aware of the current screening methods to accurately gage alcohol use in pregnant and childbearing women. PMID- 11860608 TI - Raltitrexed (Tomudex) administration in patients with relapsed metastatic colorectal cancer after weekly irinotecan/5-Fluorouracil/Leucovorin chemotherapy. AB - PURPOSE: The present study aimed at evaluating the efficacy of Raltitrexed, a specific thymidilate synthase inhibitor, in patients with advanced colorectal cancer (ACC) in relapse (>8 weeks) after a prior response or disease stabilization to first-line chemotherapy combination with lrinotecan+5 Fluorouracil (5-FU)+Leucovorin (LV). METHODS: Twenty-five patients with metastatic ACC entered; 17 males/8 females, median age 61 (range: 47-70), median Karnovsky PS: 80 (70-90), and sites of metastases; liver: 21, lung: 4, lymph nodes: 7, peritoneal: 5 and a life expectancy of at least 3 months, were entered in the present pilot study. All patients had progressed after prior chemotherapy with lrinotecan+5-FU+LV. Raltitrexed was administered at a dose of 3 mg/m2 i.v. every 21 days. RESULTS: Three patients (12%) achieved a partial response (PR), 8 (32%) had stable disease (SD), and the remaining 14 (56%) developed progressive disease (PD). Median time-to-progression (TTP) was 5.5 months (range, 2-8.5), and median overall survival (OS) 8 months (range, 4.0-12.5). Toxicity was generally mild; it consisted mainly of myelosuppression; neutropenia grade 1-2: 52%-grade 3: 28%, and anemia grade 1-2 only: 36%. Mild mucositis grade 1-2 occurred in 13.5% of patients and was the principal non-hematologic toxicity. CONCLUSION: Response to treatment with Raltitrexed is limited in patients with ACC failing after an initial response or non-progression to the weekly lrinotecan+5-FU+LV combination. However, it appears that a limited number of patients with PR/SD may derive clinical benefit, but final proof would require a randomized study. PMID- 11860609 TI - The influence of age on the female/male ratio of treated incidence rates in depression. AB - BACKGROUND: Poor data exist on the influence of psychosocial variables on the female/male ratio of depression because of the small number of cases and the resulting limited numbers of variables available for investigation. For this investigation a large number of first admitted depressed patients (N = 2599) was available which offered the unique opportunity to calculate age specific sex ratios for different marital and employment status categories. METHODS: Age and sex specific population based depression rates were calculated for first ever admissions for single year intervals. Moving averages with interpolated corrections for marginal values in the age distribution were employed. RESULTS: For the total group the female/male ratio of depression showed an inverted U shape over the life-cycle. This pattern was influenced by the group of married persons, which showed a sex-ratio of 3:1 between the age of 30-50, but ratios of around 1:1 at younger and older ages. For not married persons the female/male ratio was already around 2:1 at the age of 18 and rose to 2.5:1 in mid-life and declined to 1 at around 55. The almost parallel decline of depression rates in employed men and women resulted in a female/male ratio of about 2:1 from age 18 to age 50 and became 1 after the age of 60. The female/male ratio among the not employed was about 1, in mid-life it became negative. CONCLUSIONS: Our analyses show that the gender-gap in first admitted depressed patients is age dependent and that psychosocial factors modify the sex ratio. PMID- 11860610 TI - Current practices in managing acutely disturbed patients at three hospitals in Rio de Janeiro-Brazil: a prevalence study. AB - BACKGROUND: The medical management of aggressive and violent behaviour is a critical situation for which there is little evidence. In order to prepare for a randomised trial, due to start in the psychiatric emergency rooms of Rio de Janeiro in 2001, a survey of current practice was necessary. METHODS: A seven day survey of pharmacological management of aggressive people with psychosis in the emergency rooms of all four public psychiatric hospitals in Rio de Janeiro, Brazil. RESULTS: In one hospital data were not available. Of the 764 people with psychosis attending these ERs, 74 were given IM medication for rapid tranquillisation (9.7%, 2.1/week/100,000). A haloperidol-promethazine mix (with or without other drugs) was used for the majority of patients (83%). CONCLUSION: The haloperidol-promethazine mix, given intramuscularly for rapid tranquilization, is prevalent in Rio, where it is considered both safe and efficient. However, scientific evaluation of all pharmacological approaches to rapid tranquilization of psychotic people is inadequate or incomplete and a randomized trial of IM haloperidol-promethazine is overdue. PMID- 11860611 TI - Reliability of race assessment based on the race of the ascendants: a cross sectional study. AB - BACKGROUND: Race is commonly described in epidemiological surveys based on phenotypic characteristics. Training of interviewers to identify race is time consuming and self identification of race might be difficult to interpret. The aim of this study was to determine the agreement between race definition based on the number of ascendants with black skin colour, with the self-assessment and observer's assessment of the skin colour. METHODS: In a cross-sectional study of 50 women aged 14 years or older, from an outpatient clinic of an University affiliated hospital, race was assessed through observation and the self assignment of the colour of skin and by the number of black ascendants including parents and grandparents. Reliability was measured through Kappa coefficient. RESULTS: Agreement beyond chance between self-assigned and observed skin colour was excellent for white (0.75 95% CI 0.72-0.78) and black women (0.89 95% CI 0.71 0.79), but only good for participants with mixed colour (0.61 95% CI 0.58-0.64), resulting in a global kappa of 0.75 (95% CI 0.71-0.79). However, only a good agreement for mixed women was obtained. The presence of 3 or more black ascendants was highly associated with observed and self-assessed black skin colour. Most women self-assigned or observed as white had no black ascendants. CONCLUSIONS: The assessment of race based on the race of ascendants showed reasonable agreement with the ascertainment done by trained interviewers and with the self-report of race. This method may be considered for evaluation of race in epidemiological surveys, since it is less time-consuming than the evaluation by interviewers. PMID- 11860612 TI - Norepinephrine-evoked pain in fibromyalgia. A randomized pilot study [ISRCTN70707830]. AB - BACKGROUND: Fibromyalgia syndrome displays sympathetically maintained pain features such as frequent post-traumatic onset and stimuli-independent pain accompanied by allodynia and paresthesias. Heart rate variability studies showed that fibromyalgia patients have changes consistent with ongoing sympathetic hyperactivity. Norepinephrine-evoked pain test is used to assess sympathetically maintained pain syndromes. Our objective was to define if fibromyalgia patients have norepinephrine-evoked pain. METHODS: Prospective double blind controlled study. PARTICIPANTS: Twenty FM patients, and two age/sex matched control groups; 20 rheumatoid arthritis patients and 20 healthy controls. Ten micrograms of norepinephrine diluted in 0.1 ml of saline solution were injected in a forearm. The contrasting substance, 0.1 ml of saline solution alone, was injected in the opposite forearm. Maximum local pain elicited during the 5 minutes post-injection was graded on a visual analog scale (VAS). Norepinephrine-evoked pain was diagnosed when norepinephrine injection induced greater pain than placebo injection. Intensity of norepinephrine-evoked pain was calculated as the difference between norepinephrine minus placebo-induced VAS scores. RESULTS: Norepinephrine-evoked pain was seen in 80 % of FM patients (95% confidence intervals 56.3 - 94.3%), in 30 % of rheumatoid arthritis patients and in 30 % of healthy controls (95% confidence intervals 11.9 - 54.3) (p < 0.05). Intensity of norepinephrine-evoked pain was greater in FM patients (mean +/- SD 2.5 +/- 2.5) when compared to rheumatoid arthritis patients (0.3 +/- 0.7), and healthy controls (0.3 +/- 0.8) p < 0.0001. CONCLUSIONS: Fibromyalgia patients have norepinephrine-evoked pain. This finding supports the hypothesis that fibromyalgia may be a sympathetically maintained pain syndrome. PMID- 11860613 TI - Modeling of failure mode in knee ligaments depending on the strain rate. AB - BACKGROUND: The failure mechanism of the knee ligament (bone-ligament-bone complex) at different strain rates is an important subject in the biomechanics of the knee. This study reviews and summarizes the literature describing ligament injury as a function of stain rate, which has been published during the last 30 years. METHODS: Three modes of injury are presented as a function of strain rate, and they are used to analyze the published cases. The number of avulsions is larger than that of ligament tearing in mode I. There is no significant difference between the number of avulsions and ligament tearing in mode II. Ligament tearing happens more frequently than avulsion in mode III. RESULTS: When the strain rate increases, the order of mode is mode I, II, III, I, and II. Analytical models of ligament behavior as a function of strain rate are also presented and used to provide an integrated framework for describing all of the failure regimes. In addition, this study showed the failure mechanisms with different specimens, ages, and strain rates. CONCLUSION: There have been several a numbers of studies of ligament failure under various conditions including widely varying strain rates. One issue in these studies is whether ligament failure occurs mid-ligament or at the bone attachment point, with assertions that this is a function of the strain rate. However, over the range of strain rates and other conditions reported, there has appeared to be discrepancies in the conclusions on the effect of strain rate. The analysis and model presented here provides a unifying assessment of the previous disparities, emphasizing the differential effect of strain rate on the relative strengths of the ligament and the attachment. PMID- 11860614 TI - Effect of vitamin D on bone mineral density of elderly patients with osteoporosis responding poorly to bisphosphonates. AB - BACKGROUND: Bisphosphonates are indicated in the prevention and treatment of osteoporosis. However, bone mineral density (BMD) continues to decline in up to 15% of bisphosphonate users. While randomized trials have evaluated the efficacy of concurrent bisphosphonates and vitamin D, the incremental benefit of vitamin D remains uncertain. METHODS: Using data from the Canadian Database of Osteoporosis and Osteopenia (CANDOO), we performed a 2-year observational cohort study. At baseline, all patients were prescribed a bisphosphonate and counseled on vitamin D supplementation. After one year, patients were divided into two groups based on their response to bisphosphonate treatment. Non-responders were prescribed vitamin D 1000 IU daily. Responders continued to receive counseling on vitamin D. RESULTS: Of 449 patients identified, 159 were non-responders to bisphosphonates. 94% of patients were women. The mean age of the entire cohort was 74.6 years (standard deviation = 5.6 years). In the cohort of non-responders, BMD at the lumbar spine increased 2.19% (p < 0.001) the year after vitamin D was prescribed compared to a decrease of 0.55% (p = 0.36) the year before. In the cohort of responders, lumbar spine BMD improved 1.45% (p = 0.014) the first year and 1.11% (p = 0.60) the second year. The difference between the two groups was statistically significant the first year (p < 0.001) but not the second (p = 0.60). Similar results were observed at the femoral neck but were not statistically significant. CONCLUSION: In elderly patients with osteoporosis not responding to bisphosphonates, vitamin D 1000 IU daily may improve BMD at the lumbar spine. PMID- 11860615 TI - Rheumatoid arthritis, gold therapy, contact allergy and blood cytokines. AB - OBJECTIVE: To study the clinical and biochemical effects of a low starting dose for gold therapy in rheumatoid arthritis patients with a contact allergy to gold. METHODS: Serum cytokines were assayed before and 24 h after the first injection of gold sodium thiomalate (GSTM). RESULTS: Contact allergy to gold was found in 4 of 19 patients. Compared to gold-negative patients (starting dose: 10 mg GSTM), there was a larger increase in serum TNFalpha (p < 0.05), sTNF-R1 (NS), and IL-1 ra (p < 0.05) in gold-allergic patients. CONCLUSIONS: Cytokines are released in blood by GSTM in RA patients with gold allergy. To minimize the risk of acute adverse reactions the starting dose of GSTM should be lowered to 5 mg. Alternatively, patients should be patch-tested before gold therapy; in test positive cases, 5 mg is recommended as the first dose. PMID- 11860616 TI - Plant growth regulatory effect and insecticidal activity of the extracts of the Tree of Heaven (Ailanthus altissima L.). AB - BACKGROUND: There is an urgent need to explore and utilize naturally occurring products for combating harmful agricultural and public health pests. Secondary metabolites in the leaves of the Tree of Heaven, Ailanthus altissima L. have been reported to be herbicidal and insecticidal. The mode of action, however, of the active compounds in A. altissima are not understood. In this paper, we report the chemical characteristics of the herbicidal and insecticidal components in this tree, and will discuss the effect of light on the bioactivity of the active components. RESULTS: Extracts from the fresh leaves of A. altissima showed a strong plant germination/growth inhibitory effect in laboratory bioassays against alfalfa (Medicago sativa). The effect was dose-dependent. The growth inhibitory components were in the methylene chloride soluble fraction of the extract. The effect was greater in the light than in the dark. Other fractions had plant growth enhancing effect at lower concentrations. The extract was slightly insecticidal against yellow fever mosquito larvae (Aedes aegypti). CONCLUSIONS: The extract or its semi-purified fractions of A. altissima were strong plant growth inhibitors, therefore good candidates as potential environmentally safe and effective agricultural pest management agents. The finding that light affects the activity will be useful in the application of such natural products. PMID- 11860617 TI - Structural model of nicotinic acetylcholine receptor isotypes bound to acetylcholine and nicotine. AB - BACKGROUND: Nicotine is a psychoactive drug presenting a diverse array of biological activities, some positive, such as enhancement of cognitive performances, others negative, such as addiction liability. Ligands that discriminate between the different isotypes of nicotinic acetylcholine receptors (nAChRs) could present improved pharmacology and toxicity profile. RESULTS: Based on the recent crystal structure of a soluble acetylcholine binding protein from snails, we have built atomic models of acetylcholine and nicotine bound to the pocket of four different human nAChR subtypes. The structures of the docked ligands correlate with available biochemical data, and reveal that the determinants for isotype selectivity are relying essentially on four residues, providing diversity of the ligand binding pocket both in terms of Van der Waals boundary, and electrostatic potential. We used our models to screen in silico a large compound database and identify a new ligand candidate that could display subtype selectivity. CONCLUSION: The nAChR-agonist models should be useful for the design of nAChR agonists with diverse specificity profiles. PMID- 11860642 TI - Identification of the auditory hazards of extracorporeal shock wave lithotripsy. AB - The aim of this work was to determine the effect of extracorporeal shock wave lithotripsy (ESWL) on the hearing of both patients and staff members exposed to such treatment. It used different hearing screening instruments, and compared the sensitivity of these instruments for the detection of the earliest change in hearing induced by this procedure. The results of this study show that ESWL has a potentially hazardous effect on hearing. This effect is subtle, could only be detected by transient evoked otoacoustic emissions (TEOAE), and is manifested in some of the subjects under study as temporary subjective hearing loss and tinnitus, reflecting a state of temporary biomechanical derangement of the outer hair cells. This effect seems to be related to the frequency of exposure to ESWL. PMID- 11860643 TI - Endoscopic orbital decompression for Graves' ophthalmopathy. AB - Graves' disease may occasionally result in significant proptosis that is either cosmetically unacceptable or causes visual loss. This has traditionally been managed surgically by external decompression of the orbital bony skeleton. Trans nasal endoscopic orbital decompression is emerging as a new minimally-invasive technique, that avoids the need for cutaneous or gingival incisions. Decompression of the medial orbital wall can be performed up to the anterior wall of the sphenoid sinus. This can be combined with resection of the medial and posterior portion of the orbital floor (preserving the infra-orbital nerve). This technique produces decompression which is comparable to external techniques. We present a series of 10 endoscopic orbital decompressions with an average improvement of 4.4 mm in orbital proptosis. There was an improvement in visual acuity in all patients with visual impairment. Endoscopic orbital decompression is recommended as an alternative to traditional decompression techniques. PMID- 11860644 TI - A quantitative analysis of the intranasal delivery of topical nasal drugs to the middle meatus: spray versus drop administration. AB - The delivery of nasal drugs specifically to the middle meatus is of critical importance in the medical treatment of rhinosinusitis. In this respect, topical nasal drug administration by drops has generally been perceived to be superior to nasal sprays, although there is a lack of evidence to support this notion. This study aims to compare the intranasal delivery of nasal sprays and drops to the middle meatus in vivo, using a novel quantitative method. A surgical patty was placed in the middle meatus. Radio-labelled topical nasal drops and aqueous sprays were administered in a standardized fashion in normal volunteers (10 nasal cavities). The subsequent absorption of administered radio-labelled saline on the patty was measured using a gamma counter. A randomized prospective crossover design was used for the study. The mean percentage (range) of absorbed administered saline on the swab was 8.7 (0.3-39.5) and 9.7 (0.03-20.4) for the spray and drop administration techniques respectively (p = 0.8). Thus, there is wide variation in the delivery of topical nasal drugs and the perceived superiority of nasal drop administration, in terms of delivery to the middle meatus, may be incorrect. PMID- 11860645 TI - Growth of laryngeal glands in infants: a comparative study of the sub- and supraglottic larynx. AB - It is well established that laryngeal secretions, produced in the glandular tissue, contribute to the local defence system of the larynx. In this study, the distribution of glandular tissues in the subglottic and supraglottic larynx was examined in 55 infants, using an image analysing system. The average area of the mucosa or glandular acini at the level of the false vocal fold was almost three times larger than that of the subglottis. At both levels, the area occupied by the glands increased according to the age at death and became more evident according to the total development score (TDS) which was estimated by adding the age at death and the gestational period. The results indicate that the glandular structure of the level of the sub- or supraglottic larynx develops constantly and equally after birth. PMID- 11860646 TI - Experience with vertical partial laryngectomy with special reference to laryngeal reconstruction with cervical fascia. AB - In this paper we report our experience of vertical partial laryngectomy using the superficial cervical fascia; we describe the technique and present the functional and oncological results of this method of treatment. A total of 42 patients with squamous cell carcinoma of the true vocal folds, in stage T(1) (n = 28) or T(2) (n = 14), were treated in our department using vertical partial laryngectomy during the decade 1987-1997. Nine patients had post-operative radiotherapy. The shortest follow-up time was three years. There were six recurrences in all, four in the larynx and two in the neck. All four of the laryngeal recurrences were treated with total laryngectomy and are doing well. Both the patients with neck metastases, who were treated with neck dissection, died. Permanent tracheotomy was necessary in one patient. There were no problems with aspiration. The recurrence rate was 14 per cent, the three-year survival index was 95.2 per cent and the three-year larynx preservation index was 90 per cent. According to our experience, vertical partial laryngectomy, using the method we describe, has a good functional and oncological result for stage T(1) and T(2) tumours. PMID- 11860647 TI - Squamous cell carcinoma of the hypopharynx treated with surgery and radiotherapy. AB - A series of squamous cell carcinomas (SCC) of the hypopharynx treated with combined surgery and radiotherapy is presented to highlight the results of treatment at an early stage of disease. A retrospective mono-institutional analysis was performed on 153 previously untreated patients with SCC of the hypopharynx, seen between 1980 and 1995 at our institution. Univariate and multivariate analyses were performed using the Cox proportional hazard model. The overall five-year specific, and non-specific, disease survival rates were 68 per cent (95 per cent confidence interval, CI: 60-77) and 47 per cent (95 per cent CI: 39-56), respectively. Compared with other series, this study is characterized by treatment at an earlier stage, better prognosis, and a higher number of multiple malignancies. Twenty-two per cent of hypopharyngeal SCCs were diagnosed during the staging procedures for a different head and neck SCC and 14 per cent during the follow-up for a previous tumour. Multivariate survival analysis of clinical and pathological factors confirmed the clinical class of tumour (T) and node (N) and the nodal capsular rupture as prognosticators of disease. PMID- 11860648 TI - Results of radiotherapy with, or without, salvage surgery versus combined surgery and radiotherapy in advanced carcinoma of the hypopharynx. AB - There is considerable controversy surrounding the optimum treatment of advanced hypopharyngeal cancers. Curative radiotherapy with surgical salvage in reserve is an accepted protocol as is also a combined treatment of surgery and radiotherapy. The present study is a retrospective analysis of the survival results of 195 cases treated in a single centre. The combined surgery and radiotherapy group comprised a greater number of pyriform fossa and post-cricoid tumours whereas, the curative radiotherapy group had a higher proportion of posterior pharyngeal wall tumours. Actuarial two-year disease-free survival rates were significantly better with combined treatment when results of stage III and IV lesions (164 patients) of all sites are taken together, as compared to those obtained with curative radiotherapy without salvage (p = 0.000) or radiotherapy with surgical salvage for residual/recurrent tumours (p = 0.0021). PMID- 11860649 TI - Topical bupivacaine in paediatric day-case tonsillectomy: a prospective randomized controlled trial. AB - Post-operative pain and delayed oral intake are common reasons for failure of discharge in day-case tonsillectomy. A double blind prospective randomized study was devised to investigate the effectiveness of topical bupivacaine in reducing post-operative pain in paediatric day-case tonsillectomy. Ninety-nine patients aged between three and 16 years were recruited into the study. One group received bupivacaine soaked swabs tightly packed in their tonsillar fossae while the control group received saline-soaked swabs. The bupivacaine group was found to drink (p < 0.001) and eat (p = 0.006) earlier than the control group. The pain scores at one (p < 0.001), three (p < 0.001) and six (p < 0.001) hours post operatively were also found to be lower in the bupivacaine group than the control group. We conclude that topical bupivacaine has a role in facilitating recovery in day-case tonsillectomy in children. PMID- 11860650 TI - A method for providing a unilateral pressure dressing in the neck. AB - The application of a reliable adequate pressure dressing to one side of the neck has always proved difficult, because of the risk of occluding the airway and the vasculature. We describe a method of maintaining well-localized pressure to one side of the neck without undue pressure on the vasculature and the airway by using a stiff cervical collar. This method was used successfully in the conservative management of neck haematoma in two patients. PMID- 11860651 TI - B-cell lymphoma of the external auditory meatus. AB - A 53-year-old female presented with a painful swelling within her external auditory meatus. Biopsies revealed this to be a B-cell lymphoma and she underwent surgical treatment followed by chemotherapy. This is the first reported case of non-Hodgkin's lymphoma of the external auditory meatus in an human immunodeficiency virus (HIV)-negative patient. PMID- 11860652 TI - Pre-operative embolization of auricular arteriovenous fistula. AB - Treatment of auricular arteriovenous fistula (AVF) is a challenge with surgery being the preferred option until now. We present three cases of auricular AVFs who underwent pre-operative embolization and its outcome on surgery. Three patients were diagnosed to have auricular AVF by angiography. All three patients underwent pre-operative embolization with n-butyl 2-cyanoacrylate after which they underwent surgical resection of the AVF. Pre-operative embolization resulted in significant devascularization of the AVF thus leading to near bloodless and clean surgery. Pre-operative embolization of auricular AVFs is a good treatment option, leading to significantly reduced blood loss during surgical excision. PMID- 11860653 TI - Bilateral simultaneous facial nerve palsy. AB - Bilateral simultaneous facial nerve palsy is an extremely rare clinical entity with Bell's palsy responsible for a mere 20 per cent of cases. It is, therefore, important that clinicians are aware of the differential diagnosis when evaluating a case. PMID- 11860654 TI - Plexiform neurofibroma of the larynx in a child. AB - A case of a female child of six years of age with a plexiform neurofibroma of the larynx caused by von Recklinghausen's disease is presented. Laryngeal involvement in neurofibromatosis type 1 (NF1) is rare and only 19 paediatric cases have been reported. The tumour was biopsied and lateral pharyngotomy with supraglottic hemilaryngectomy was performed in order to relieve obstructive symptoms. Problems related to this unusual tumour localization are discussed and a review of the literature is presented. PMID- 11860655 TI - Basaloid squamous cell carcinoma of the larynx. AB - Most of the malignancies arising from the upper respiratory system are squamous cell carcinomas (SCC). Basaloid squamous cell carcinoma (BSCC) is a rarely seen high grade variant of SCC that carries a poor prognosis. In this article, we present a case of BSCC of the larynx. In addition, we provide a review of the relevant literature along with a discussion of the differential diagnosis. PMID- 11860656 TI - A rare case of dysphagia: hypopharyngeal amyloidosis masquerading as a post cricoid tumour. AB - Amyloidoses are a group of disorders in which deposition of abnormal amounts of protein complexes (amyloid) occurs in a variety of tissues. The upper aerodigestive tract may be affected, particularly the larynx, but hypopharyngeal involvement is rarely reported. We present a unique case of amyloidosis of the post-cricoid region causing dysphagia. This case report highlights the need for otolaryngologists to consider the possibility of submucosal amyloid deposition, in the absence of mucosal lesions, in patients who present with dysphagia secondary to an obstructive lesion of the post cricoid region. PMID- 11860657 TI - Staged endoscopic operation for large pituitary adenomas. AB - The advantages of endoscopic transnasal pituitary surgery include a panoramic view of the area and easy reoperation. Based on this, we performed a planned staged operation in two cases of large pituitary adenomas, extending to the suprasellar lesion, using the endoscopic transnasal trans-sphenoidal approach via both nostrils. In the initial operation, a tumour mass was from the intrasellar region, however, that part of it in contact with the tumour capsule was left intact for prevention of peri-operative injury. The residual mass descended into the sellar floor at the second operation, and was easily resected with no complications. A staged operation by the endoscopic transnasal approach, although it requires two operations, is thought to be a safe and efficient procedure for the removal of large pituitary tumours. PMID- 11860658 TI - Shock wave lithotripsy of a parotid duct calculus. AB - Extracorporeal shock wave lithotripsy, a relatively new non-invasive method of treating salivary gland calculi, is rarely used in UK hospitals. We present a case of parotid duct calculus treated successfully by lithotripsy in a United Kingdom (UK) district General Hospital. PMID- 11860659 TI - Hypoglossal schwannoma in the submandibular space. AB - Most schwannomas of the hypoglossal nerve originate from the intracranial portion, but they may extend extracranially. Solitary and extracranial schwannomas are extremely rare. We report a case of submandibular hypoglossal schwannoma along with its clinical course and management. PMID- 11860660 TI - Unusual solitary osteochondroma of the mandibular ramus. AB - Mandibular osteochondromas, although rare, predominantly involve the condyloid and coronoid processes. An extremely rare case arising from the inner aspect of the gonion is reported here. Only one such case has been reported in the literature so far. Despite being very slow growing and having a benign course, histopathological examination showed reactive changes that may be confused with sarcomatous transformation. We advocate surgical excision using a combined external and internal approach followed by regular post-operative review. The relevant literature has been reviewed but various surgical approaches as advocated in the past have not been found suitable. PMID- 11860661 TI - Thrombosed varicose vein in the sternocleidomastoid muscle. AB - A thrombosed varicose vein arising in the neck is quite rare. A 68-year-old Japanese male was referred to our hospital because of a mass in his left upper neck. The mass did not change in size in response to strain resulting from Valsalva's manouvre. Magnetic resonance imaging (MRI) showed iso-signal intensity of the mass on T1-weighted images and a target-like signal arrangement (concentric hyper-, hypo-, hyper-signal intensity from outside to inside) on T2 weighted images. Surgical excision revealed that the tumour arose from the intramuscular small vein in the sternocleidomastoid muscle. The pathological examination revealed the mass to be a thrombosed varicose vein with capillarization in the dilated vein wall. The de-oxygenation and degradation of haemoglobin were thought to be responsible for these characteristic MRI findings. The concentric signal distribution on MRI strongly suggested this pathology. PMID- 11860662 TI - The use of maggots in head and neck necrotizing fasciitis. AB - Historically maggots have been used to clean necrotic war wounds. With the ready availability of sterile maggots, they are being used increasingly in surgical practice. Although maggots have most frequently been employed in the management of necrotic wounds and ulcers involving the lower limb, we have identified a particularly useful application in the head and neck and describe a case in which maggots played a significant part in the successful treatment of a florid necrotizing cervical fasciitis in a patient, who was unfit for repeated surgical debridement. PMID- 11860663 TI - An unusual case of surgical emphysema in the neck following sport injury. AB - Surgical emphysema of the neck following sport injury without direct trauma to the neck is uncommon. We report a case of a flexion-hyperextension injury causing an air leak to the soft tissues of the neck. Diagnosis, management and potential mechanisms are discussed. Patients may initially present with minimal symptoms, but their condition may deteriorate rapidly or insidiously. In the absence of respiratory compromise, conservative management is appropriate. PMID- 11860664 TI - Pneumomediastinum following Ecstasy (methylenedioxymetamphetamine, MDMA) ingestion in two people at the same 'rave'. AB - Ecstasy is a class A controlled drug often consumed by the young population for recreational purposes. Documented complications of its use include hyperpyrexia, disseminated intravascular coagulation (DIC), renal failure and rhabdomyolysis. We report on two patients who developed pneumomediastinum after Ecstasy abuse. Both patients obtained and consumed the drug at the same establishment and presented to the same hospital within half an hour. The possible pathogenesis of this complication are discussed and the literature reviewed. Pneumomediastinum should be recognized as a possible complication of Ecstasy use. Conservative management is appropriate. PMID- 11860666 TI - Patient-rated versus clinician-rated side effects of drug treatment in schizophrenia. Clinical validation of a self-rating version of the UKU Side Effect Rating Scale (UKU-SERS-Pat). AB - A self-rating version of the UKU Side Effect Rating Scale has been developed. The present study examines the agreement between patients' self-assessment of side effects and the attending clinicians' ratings. The patient sample consisted of 63 patients with schizophrenia under maintenance treatment with risperidone, clozapine or classical antipsychotics. Approximately two thirds of the patients used concomitant medication with e.g. benzodiazepines, SSRIs, anticholinergics. Most inter-correlations between scores for single, corresponding items, subscores of Psychic, Neurological, Autonomic and Other side effects, as well as the Total Score from the patient version of the UKU Side Effect Self Rating Scale (UKU-SERS Pat) and the clinician version (UKU-SERS-Clin) were found to be statistically significant. Patients reported side effects more frequently and or rated symptoms more severe than the clinicians. The results support the validity of the SERS-Pat and suggest that patient rated side effects may provide important clinical information not detected by clinician rated interviews. Such information can be utilised both in clinical investigations, in development of treatment programs and for individual patients in clinical practice. PMID- 11860667 TI - Assessment of patient satisfaction with psychiatric care. Development and clinical evaluation of a brief consumer satisfaction rating scale (UKU-ConSat). AB - Quality management and quality control of health services has become increasingly important. Central to the concept of quality of care is the patient's (the health care consumer's) own view of the care provided and the treatment outcome. UKU (Udvalg for Kliniske Undersogelser; that is, Committee for Clinical Trials), a working group within the Scandinavian Society for Psychopharmacology (SSP), has designed a brief consumer satisfaction rating scale, the UKU-ConSat. Based on an extensive literature review, a number of principles related to content, assessment, interview techniques, documentation and standardisation steered the design of the rating scale. UKU-ConSat consists of six items related to the structure and process of treatment care, and two items related to outcome and well-being. A manual accompanies the rating scale with guidelines for how to solicit information from the patient and how to rate each item. A first field trial of the rating scale in 135 inpatients at multiple clinical sites in Finland and Sweden showed that it could be applied to several relevant patient categories (psychotic, affective, neurotic, organic and alcohol and substance abuse disorders). According to both patients and staff the rating scale promises to become useful both for research and for improvement of routine psychiatric services. PMID- 11860668 TI - Consumer satisfaction in schizophrenia. A 2-year randomized controlled study of two community-based treatment programs. AB - Quality management and quality control of health services have become increasingly important. Central to the concept of quality of care is the consumer's (the patient's) own view of the care provided. UKU (Udvalg for Kliniske Undersogelser), a task force within the Scandinavian Society for Psychopharmacology, has designed a brief consumer satisfaction rating scale, the UKU-ConSat. The UKU-ConSat rating scale was applied in a randomised comparative two-year study of two community-based treatment programs. The study included 84 patients with schizophrenic disorders according to DSM-IV. The major finding was a significantly increased consumer satisfaction with the new community-based program "Integrated Care" in comparison with best-established practice, called "Rational Rehabilitation." Thus, the new UKU-ConSat rating scale can detect differences, not only for in-patients as has previously been shown, but also between programs in community care settings. By per item analysis it was possible to discriminate between various elements of consumer satisfaction. There were significant correlations between the UKU-ConSat total scores and other clinical outcome domains, viz. symptoms, social function, and patient as well as carer's distress. To conclude, the UKU-ConSat rating scale is suitable for quality management and monitoring of treatment programs in mental health services. PMID- 11860669 TI - AIDS-related systemic non-Hodgkin's lymphoma at a large community program. AB - The introduction of triple antiretroviral therapy has led to reductions in opportunistic diseases in HIV-infected patients. However, little is known of the effect of this therapy on the clinical and pathological features and the outcome of patients with AIDS-related systemic non-Hodgkin's lymphoma (NHL). We examined the incidence and clinical manifestations of HIV-infected patients with systemic NHL at the Harris County Hospital District and Veterans Affairs Medical Center (Houston, TX). Between January 1996 and December 2000, 76 cases of systemic NHL were diagnosed in 3655 HIV-infected patients. Three groups of patients diagnosed with systemic NHL were identified according to their history of antiretroviral therapy: treatment naive (n = 20), dual nucleoside (n = 22), and triple antiretroviral drug-treated patients (n = 34). The median duration of antiretroviral therapy before the diagnosis of systemic NHL in the triple antiretroviral and dual nucleoside treatment groups was 12 versus 8 months (p < 0.0004). Thirty-five percent of patients (12 of 34) in the triple treatment group had an HIV RNA viral load of <400 copies/ml and their median CD4+ cell count was 301 cells/mm(3) (range, 46 to 667 cells/mm(3)) at the time of diagnosis of systemic NHL. More patients treated with triple antiretroviral therapy received complete courses of chemotherapy as compared with the other two groups (p = 0.013). However, the overall survival did not differ significantly among the three groups of patients. These data suggest that AIDS-related systemic NHL continues to occur even in patients treated with triple antiretroviral therapy. In addition, this opportunistic malignancy is associated with significant mortality. Therefore, it is necessary to develop a better understanding of the pathogenesis of this disease. PMID- 11860670 TI - Underestimation of HIV type 1 drug resistance mutations: results from the ENVA-2 genotyping proficiency program. AB - Current recommendations indicate the use of HIV-1 drug resistance genotyping in the treatment of HIV-1 infection, primarily on treatment failure, and in specific instances also before the initiation of therapy. As such, HIV-1 genotyping is becoming a standard of care parameter in HIV-1 treatment monitoring and a rapidly increasing number of laboratories now use this technology routinely. A study of proficiency, using the ENVA-2 panel, was performed to evaluate the current HIV-1 resistance genotyping quality in 34 laboratories from different parts of the world. The results demonstrated extensive interlaboratory variation in the quality of genotyping and a significant underestimation of resistance mutations, even in samples expressing pure mutant genotype. The level of variation could not be attributed to the sequencing technology used and was therefore considered to be laboratory associated. The direct clinical consequences of this may be inadequate treatment of HIV-1-infected individuals and a more rapid exhaustion of therapeutic options for the patients. Drug resistance mutations are frequently missed. Therefore, quality control programs are urgently needed. Until these are widely implemented, clinicians must consider this issue and interpret the reported genotyping results with caution. PMID- 11860671 TI - Telomerase activity of human peripheral blood mononuclear cells in the course of HTLV type 1 infection in vitro. PMID- 11860672 TI - Monoclonally integrated HTLV type 1 in epithelial cancers from rabbits infected with an HTLV type 1 molecular clone. AB - In addition to T cell leukemias and lymphomas, human T cell leukemia virus type 1 (HTLV-1) infection has been associated with nonhematologic malignancies and described as the cause of one case of small-cell lung carcinoma. Infected primary epithelial cells have been isolated from sweat gland and oral mucosae of HTLV-1 infected human patients. In the present study, epithelial neoplasms developed in two rabbits experimentally infected with a molecular clone of HTLV-1 (strain K30p). Serologic detection of anti-HTLV-1 and isolation of virus from blood lymphocytes at multiple time points postinjection established a course of chronic asymptomatic infection in both. One rabbit, infected for 5.5 years after intramuscular injection of HTLV-1 DNA, developed a thymoma having features of medullary differentiation. HTLV-1 provirus was detected in both thymocytes and neoplastic epithelium isolated discretely from the thymoma by laser capture microdissection. These findings provide the first experimental evidence of HTLV-1 disease after infection by HTLV-1 DNA injection. Endometrial adenocarcinoma occurred in a second rabbit 2.5 years after its inoculation with cell-associated virus. In this second case, an epithelial cell line derived ex vivo from a metastatic lesion produced virus in culture. In tumors from each of the two rabbits, the neoplastic epithelium was infected and harbored monoclonally integrated HTLV-1 provirus. Although monoclonal provirus integration alone does not establish retroviral cause of carcinogenesis unequivocally, these and other accumulating data indicate that there may be a role for HTLV-1 in diseases associated with infection of epithelia, including some epithelial cancers. PMID- 11860673 TI - Cellular factors influence the binding of HIV type 1 to cells. AB - The goal of this study was to determine the importance of cellular factors for binding of HIV to cells. HIV primary isolates (PIs) produced in peripheral blood mononuclear cells (PBMCs) bound at relatively high levels to PBMCs but at low levels to cell lines, whereas T cell line-adapted (TCLA) virus produced in the H9 T cell line bound at high levels to both cell lines and PBMCs. Expression of CD4 in CD4-negative cells or blocking CD4 with antibody on CD4-positive cells did not affect virus binding. Blocking of gp120/gp41 with antibodies or a lack of expression of gp120/gp41 in virus particles also did not affect virus binding. However, the cell type from which virus was produced did affect virus binding. Thus, the binding pattern of TCLA virus shifted to that of a PI virus when produced in PBMCs. A PI binding pattern also occurred when a cloned TCLA virus (NL4-3) was produced in PBMCs, indicating that the virus-producing cell type has more of an effect on virus binding than the virus strain. These experiments show that both the virus-producing cell and the target cell have a major influence on HIV binding and suggest that host cell factors incorporated into virions are important for virus binding. PMID- 11860674 TI - Evaluation of HIV type 1 group O isolates: identification of five phylogenetic clusters. AB - HIV-1 group O strains have a level of genetic diversity similar to that of strains in group M; however, group O has not been readily classified into genetic subtypes. Phylogenetic classification of group O has been hindered by the limited sequence information available. To facilitate phylogenetic analysis, we sequenced the gag p24 (693 nt), pol p32 (864 nt), and env gp160 (approximately 2700 nt) genes from 39 group O-infected specimens. These specimens include 32 plasma samples collected in Cameroon between 1996 and 1999, 2 specimens collected in the United States, and 5 infections previously isolated in Equatorial Guinea. Phylogenetic analysis of HIV-1 group O sequences resulted in the identification of five clusters that are maintained across gag, pol, and env, generally supported by high bootstrap values, and approximately equidistant from each other. In addition to the group O clusters, several isolates branch independently and are equidistant from the other group O isolates. Cluster I comprises greater than 50% of the group O isolates and is a diverse set of isolates that is subdivided into subclusters. The average intra-, sub-, and intercluster distances for group O are similar to the corresponding distances for group M subtypes. The five group O clusters have characteristics similar to those of group M subtypes. Thus the data presented may form the basis for classification of group O into subtypes. However, full-length genomes representing each group O cluster will be required to formalize a group O subtype classification. PMID- 11860675 TI - Phosphorylation of Vpr regulates HIV type 1 nuclear import and macrophage infection. AB - Viral protein R (Vpr) of human immunodeficiency virus type 1 (HIV-1) is a small accessory protein that regulates nuclear import of the viral preintegration complex and facilitates infection of nondividing cells, such as macrophages. Studies demonstrated that a fraction of Vpr molecules is phosphorylated in the virions and in HIV-1-infected cells, but the role of phosphorylation in nuclear import activity of Vpr has not been established. We found that Vpr is phosphorylated predominantly on the serine residue in position 79, and mutations affecting Vpr phosphorylation significantly attenuated viral replication in macrophages, but not in activated T lymphocytes or cell lines. The replication defect was mapped by polymerase chain reaction analysis to the step of nuclear import. These results suggest that phosphorylation of Vpr regulates its activity in the nuclear import of the HIV-1 preintegration complex. PMID- 11860676 TI - Methods of using interleukin 2 to enhance HIV-specific immune responses. AB - Interleukin 2 (IL-2) in combination with highly active antiretroviral therapy (HAART) can significantly increase CD4+ T cell counts but does not improve HIV specific T cell-mediated immune responses that are associated with the control of viral replication. To characterize the immunomodulatory activity of IL-2 in HIV infected individuals we studied the virus-specific immune response (VIR) by intracellular cytokine expression (interferon gamma, IFN-gamma) after mimicking HIV rebound in peripheral blood mononuclear cells (PBMCs). We found that the whole virus used as HIV antigen was able to activate HIV-specific T cells in the presence of a low concentration (50 IU/ml) of IL-2. Interestingly, increasing concentrations of IL-2 (400 or 1000 IU/ml) in combination with the same amount of HIV doubled the number of HIV-specific T cells. These cells were functionally intact because all of the IFN-gamma-producing CD8+ T cells contained perforin, a marker for cytotoxic T lymphocytes (CTLs). Induction of HIV-specific immune responses by IL-2 was not detected in the absence of HIV antigen both in vitro and in patients treated with HAART, indicating that IL-2 can amplify HIV-specific T cells in the presence of HIV antigen. Therefore, a combination of IL-2 with either structured treatment interruption, which results in a controlled viral load rebound, or with therapeutic vaccination is expected to improve HIV-specific T cell-mediated immune responses. PMID- 11860678 TI - High similarity between reverse-oriented sequences from HIV and foamy virus envelope glycoproteins. PMID- 11860677 TI - Quantitative detection of increasing HIV type 1 antibodies after seroconversion: a simple assay for detecting recent HIV infection and estimating incidence. AB - We have devised a simple enzyme immunoassay (EIA) that detects increasing levels of anti-HIV IgG after seroconversion and can be used for detecting recent HIV-1 infection. Use of a branched peptide that included gp41 immunodominant sequences from HIV-1 subtypes B, E, and D allowed similar detection of HIV-specific antibodies among various subtypes. Because of the competitive nature of the capture EIA, a gradual increase in the proportion of HIV-1-specific IgG in total IgG was observed for 2 years after seroconversion. This was in contrast to results obtained with the conventional EIA using the same antigen in solid phase, which plateaus soon after seroconversion. The assay was used to test 622 longitudinal specimens from 139 incident infections in the United States (subtype B) and in Thailand (subtypes B and E). The assay was also performed with an additional 8 M urea incubation step to assess the contribution of high-avidity antibodies. Normalized optical density (OD-n) was calculated (ODspecimen/ODcalibrator), using a calibrator specimen. An incremental analysis indicated that a cutoff of 1.0 OD-n and a seroconversion period of 160 days offered the best combination of sensitivity and specificity for classifying incident or long-term infections. The urea step increased the seroconversion period to 180 days with similar sensitivity and specificity. Separate analysis of B and E subtype specimens yielded the same optimal OD-n threshold and similar seroconversion periods. The assay was further validated in African specimens (subtypes A, C, and D) where the observed incidence was within 10% of the expected incidence. This assay should be useful for detecting recent HIV-1 infection and for estimating incidence among diverse HIV-1 subtypes worldwide. PMID- 11860679 TI - What geometric visual hallucinations tell us about the visual cortex. AB - Many observers see geometric visual hallucinations after taking hallucinogens such as LSD, cannabis, mescaline or psilocybin; on viewing bright flickering lights; on waking up or falling asleep; in "near-death" experiences; and in many other syndromes. Kluver organized the images into four groups called form constants: (I) tunnels and funnels, (II) spirals, (III) lattices, including honeycombs and triangles, and (IV) cobwebs. In most cases, the images are seen in both eyes and move with them. We interpret this to mean that they are generated in the brain. Here, we summarize a theory of their origin in visual cortex (area V1), based on the assumption that the form of the retino-cortical map and the architecture of V1 determine their geometry. (A much longer and more detailed mathematical version has been published in Philosophical Transactions of the Royal Society B, 356 [2001].) We model V1 as the continuum limit of a lattice of interconnected hypercolumns, each comprising a number of interconnected iso orientation columns. Based on anatomical evidence, we assume that the lateral connectivity between hypercolumns exhibits symmetries, rendering it invariant under the action of the Euclidean group E(2), composed of reflections and translations in the plane, and a (novel) shift-twist action. Using this symmetry, we show that the various patterns of activity that spontaneously emerge when V1's spatially uniform resting state becomes unstable correspond to the form constants when transformed to the visual field using the retino-cortical map. The results are sensitive to the detailed specification of the lateral connectivity and suggest that the cortical mechanisms that generate geometric visual hallucinations are closely related to those used to process edges, contours, surfaces, and textures. PMID- 11860680 TI - An amplitude equation approach to contextual effects in visual cortex. AB - A mathematical theory of interacting hypercolumns in primary visual cortex (V1) is presented that incorporates details concerning the anisotropic nature of long range lateral connections. Each hypercolumn is modeled as a ring of interacting excitatory and inhibitory neural populations with orientation preferences over the range 0 to 180 degrees. Analytical methods from bifurcation theory are used to derive nonlinear equations for the amplitude and phase of the population tuning curves in which the effective lateral interactions are linear in the amplitudes. These amplitude equations describe how mutual interactions between hypercolumns via lateral connections modify the response of each hypercolumn to modulated inputs from the lateral geniculate nucleus; such interactions form the basis of contextual effects. The coupled ring model is shown to reproduce a number of orientation-dependent and contrast-dependent features observed in center-surround experiments. A major prediction of the model is that the anisotropy in lateral connections results in a nonuniform modulatory effect of the surround that is correlated with the orientation of the center. PMID- 11860681 TI - Derivation of the visual contrast response function by maximizing information rate. AB - A graph of neural output as a function of the logarithm of stimulus intensity often produces an S-shaped function, which is frequently modeled by the hyperbolic ratio equation. The response of neurons in early vision to stimuli of varying contrast is an important example of this. Here, the hyperbolic ratio equation with a response exponent of two is derived exactly by considering the balance between information rate and the neural costs of making that information available, where neural costs are a function of synaptic strength and spike rate. The maximal response and semisaturation constant of the neuron can be related to the stimulus ensemble and therefore shift accordingly to exhibit contrast gain control and normalization. PMID- 11860682 TI - A Bayesian framework for sensory adaptation. AB - Adaptation allows biological sensory systems to adjust to variations in the environment and thus to deal better with them. In this article, we propose a general framework of sensory adaptation. The underlying principle of this framework is the setting of internal parameters of the system such that certain prespecified tasks can be performed optimally. Because sensorial inputs vary probabilistically with time and biological mechanisms have noise, the tasks could be performed incorrectly. We postulate that the goal of adaptation is to minimize the number of task errors. This minimization requires prior knowledge of the environment and of the limitations of the mechanisms processing the information. Because these processes are probabilistic, we formulate the minimization with a Bayesian approach. Application of this Bayesian framework to the retina is successful in accounting for a host of experimental findings. PMID- 11860684 TI - Activity-dependent development of axonal and dendritic delays, or, why synaptic transmission should be unreliable. AB - Systematic temporal relations between single neuronal activities or population activities are ubiquitous in the brain. No experimental evidence, however, exists for a direct modification of neuronal delays during Hebbian-type stimulation protocols. We show that in fact an explicit delay adaptation is not needed if one assumes that the synaptic strengths are modified according to the recently observed temporally asymmetric learning rule with the downregulating branch dominating the upregulating branch. During development, slow, unbiased fluctuations in the transmission time, together with temporally correlated network activity, may control neural growth and implicitly induce drifts in the axonal delays and dendritic latencies. These delays and latencies become optimally tuned in the sense that the synaptic response tends to peak in the soma of the postsynaptic cell if this is most likely to fire. The nature of the selection process requires unreliable synapses in order to give successful synapses an evolutionary advantage over the others. The width of the learning function also determines the preferred dendritic delay and the preferred width of the postsynaptic response. Hence, it may implicitly determine whether a synaptic connection provides a precisely timed or a broadly tuned "contextual" signal. PMID- 11860683 TI - Analysis of oscillations in a reciprocally inhibitory network with synaptic depression. AB - We present and analyze a model of a two-cell reciprocally inhibitory network that oscillates. The principal mechanism of oscillation is short-term synaptic depression. Using a simple model of depression and analyzing the system in certain limits, we can derive analytical expressions for various features of the oscillation, including the parameter regime in which stable oscillations occur, as well as the period and amplitude of these oscillations. These expressions are functions of three parameters: the time constant of depression, the synaptic strengths, and the amount of tonic excitation the cells receive. We compare our analytical results with the output of numerical simulations and obtain good agreement between the two. Based on our analysis, we conclude that the oscillations in our network are qualitatively different from those in networks that oscillate due to postinhibitory rebound, spike-frequency adaptation, or other intrinsic (rather than synaptic) adaptational mechanisms. In particular, our network can oscillate only via the synaptic escape mode of Skinner, Kopell, and Marder (1994). PMID- 11860686 TI - Sparse on-line gaussian processes. AB - We develop an approach for sparse representations of gaussian process (GP) models (which are Bayesian types of kernel machines) in order to overcome their limitations for large data sets. The method is based on a combination of a Bayesian on-line algorithm, together with a sequential construction of a relevant subsample of the data that fully specifies the prediction of the GP model. By using an appealing parameterization and projection techniques in a reproducing kernel Hilbert space, recursions for the effective parameters and a sparse gaussian approximation of the posterior process are obtained. This allows for both a propagation of predictions and Bayesian error measures. The significance and robustness of our approach are demonstrated on a variety of experiments. PMID- 11860685 TI - Impact of geometrical structures on the output of neuronal models: a theoretical and numerical analysis. AB - What is the difference between the efferent spike train of a neuron with a large soma versus that of a neuron with a small soma? We propose an analytical method called the decoupling approach to tackle the problem. Two limiting cases-the soma is much smaller than the dendrite or vica versa-are theoretically investigated. For both the two-compartment integrate-and-fire model and Pinsky-Rinzel model, we show, both theoretically and numerically, that the smaller the soma is, the faster and the more irregularly the neuron fires. We further conclude, in terms of numerical simulations, that cells falling in between the two limiting cases form a continuum with respect to their firing properties (mean firing time and coefficient of variation of inter-spike intervals). PMID- 11860687 TI - Orthogonal series density estimation and the kernel eigenvalue problem. AB - Kernel principal component analysis has been introduced as a method of extracting a set of orthonormal nonlinear features from multivariate data, and many impressive applications are being reported within the literature. This article presents the view that the eigenvalue decomposition of a kernel matrix can also provide the discrete expansion coefficients required for a nonparametric orthogonal series density estimator. In addition to providing novel insights into nonparametric density estimation, this article provides an intuitively appealing interpretation for the nonlinear features extracted from data using kernel principal component analysis. PMID- 11860688 TI - Natural discriminant analysis using interactive Potts models. AB - Natural discriminant analysis based on interactive Potts models is developed in this work. A generative model composed of piece-wise multivariate gaussian distributions is used to characterize the input space, exploring the embedded clustering and mixing structures and developing proper internal representations of input parameters. The maximization of a log-likelihood function measuring the fitness of all input parameters to the generative model, and the minimization of a design cost summing up square errors between posterior outputs and desired outputs constitutes a mathematical framework for discriminant analysis. We apply a hybrid of the mean-field annealing and the gradient-descent methods to the optimization of this framework and obtain multiple sets of interactive dynamics, which realize coupled Potts models for discriminant analysis. The new learning process is a whole process of component analysis, clustering analysis, and labeling analysis. Its major improvement compared to the radial basis function and the support vector machine is described by using some artificial examples and a real-world application to breast cancer diagnosis. PMID- 11860689 TI - Mycoplasma genitalium -- an up-date. AB - Mycoplasma genitalium was first isolated from men with non-gonococcal urethritis (NGU) more than 20 years ago. Use of polymerase chain reaction technology has shown it to be a cause of acute NGU and probably chronic NGU, almost independently of Chlamydia trachomatis, but there is no substantial evidence that it causes acute or chronic prostatitis. In women, M. genitalium is not associated with bacterial vaginosis, but it is strongly associated with cervicitis and endometritis and serologically with salpingitis and tubal factor infertility. Further studies may show M. genitalium to be associated, perhaps causally, with epididymoorchitis, neonatal disease and reactive arthritis. Furthermore, its potential for enhancing HIV transmission needs to be explored. M. genitalium is susceptible to various broad-spectrum antibiotics, but M. genitalium-associated diseases are probably best treated with azithromycin. PMID- 11860690 TI - Estimating HIV-1 transmission efficiency through unsafe medical injections. AB - The transmission efficiency of HIV-1 through unsafe medical injections can be estimated from seroconversions among health care workers (HCWs) after percutaneous exposures and documented iatrogenic outbreaks. Data from a case control study of seroconversion after percutaneous exposures among US and European HCWs shows an average rate of seroconversion after deep injuries - arguably comparable to unsafe injections - of 2.3%. Information from an iatrogenic HIV outbreak in a Romanian orphanage suggests a transmission efficiency of 2-7%. Even these estimates may be too low to explain the rapid spread of HIV among Russian and Libyan children in iatrogenic outbreaks discovered in 1988 and 1998, respectively. In countries in sub-Saharan Africa and elsewhere with high HIV prevalence and large numbers of unsafe injections, personal risk as well as the share of the HIV epidemic associated with unsafe injections may be an order of magnitude higher that many experts have supposed. PMID- 11860691 TI - Sexual behaviour in a low-income high school setting in Stockholm. AB - Teenage abortions and sexually transmitted infections (STIs) are increasing in Sweden, and risky adolescent sexual behaviour is an issue of major concern. The aim of the present cross-sectional questionnaire-based study was to investigate adolescent sexual experience in a low-income high school setting in Stockholm. Among 340 students, 258 participated, response rate=75.9%, mean age=17 years. Homo- or bisexual preference was reported by 11.6% (29/250). Experience of oral sex was reported by 66.3% (163/246), vaginal intercourse by 55.5% (141/254), and anal sex by 10.0% (25/250), with no gender-specific differences. Five or more coital partners were reported by 29.8% (34/114). Condom use at first and most recent intercourse was reported by 61.7% (87/141) and 42.6% (60/141), respectively, and condom breakage by 39.5% (34/86). Truancy, smoking, alcohol and drug use were factors associated with coital experience. Sexual abuse was reported by 2.2% (3/134) of the boys and 13.0% (16/123) of the girls. PMID- 11860692 TI - Are sexually transmitted infections associated with scabies? AB - Sexually transmitted infections (STIs) have been previously reported to be associated with scabies in sexually active people. The UK national guidelines on STIs recommend screening for other STIs in patients with scabies attending a genitourinary medicine (GUM) clinic. However, there is a paucity of literature on this association, which we reviewed. We looked at the sexual behaviour and the coincidence of STIs in patients with scabies attending a GUM clinic and in inmates from a young offender institution (YOI) attending between 1 January 1998 and 31 December 1999. A total of 47 patients with scabies were identified in the GUM clinic population during this period and 15 patients in the YOI group. Consecutive new and rebooked sex-matched patients served as a control group. Among the GUM clinic attendees 36% of patients with scabies had multiple sexual partners in the preceding 3 months compared with 21% in the control group (P=0.11). Thirty-four per cent of patients with scabies had a previous history of STI vs 17% in the control group (P=0.058). The number of patients with coexistent STIs was not significantly different in both groups (49% vs 44%, P=0.68). The number of patients in the YOI group was too small for a statistical analysis. This study showed that patients with scabies had comparable rates of STIs as with the GUM clinic population. It therefore supports the national guidelines in recommending screening for other STIs in patients with scabies. PMID- 11860693 TI - Evaluation of a rapid test for the detection of antibodies to human immunodeficiency virus type 1 and 2 in the setting of multiple transmitted viral subtypes. AB - Rapid HIV testing is an appealing strategy in the approach to HIV diagnosis in developing countries. Concern has been raised about the use of these tests in the setting of multiple transmitted HIV subtypes. We sought to compare the OraQuick(TM) HIV-1/2 Test, a qualitative immunochromatographic test for the detection of antibodies to HIV-1 and HIV-2 using stored sera, with a conventional enzyme immunoassay (EIA)/Western blot (WB) algorithm. The study design used was a blinded retrospective study. Samples were collected on patients attending sexually transmitted disease clinics and tuberculosis clinics in Kinshasa, the Democratic Republic of Congo and included 72 known HIV seropositive and 131 known HIV seronegative subjects. All 72 known HIV seropositive samples were positive by OraQuick and all 131 known HIV seronegative samples were negative by OraQuick resulting in 100% sensitivity and specificity. We conclude that the OraQuick rapid HIV-1/2 test performs well in the setting of diverse HIV viral subtypes. PMID- 11860694 TI - The sac-4 gene of Neisseria gonorrhoeae correlates with gonococcal subtype not co existing chlamydial infection. AB - We examined the hypothesis that the sac-4 gene of Neisseria gonorrhoeae varies with gonococcal subtype and that this could account for an earlier report that sac-4 increased the likelihood of co-infection with Chlamydia trachomatis. A polymerase chain reaction (PCR) assay was used to determine the prevalence of sac 4 in 435 gonococcal isolates. The prevalence of sac-4 was analysed in relation to chlamydial co-infection, serovar, auxotype, gender and sexual orientation. Although the prevalence of sac-4 was higher in isolates from patients with chlamydial co-infection (55%) than in those without chlamydial co-infection (42%) the difference was not significant (P<0.05). Statistically significant differences in association with sac-4 were, however, shown between various serovars and auxotypes. Dual classification based on auxotype/serovar (A/S) classes showed highly significant differences in sac-4 prevalence between groups: 95% in NR/1B18 and 8% in P/1B2 (P<0.001). Sac-4 was also significantly less common (P<0.05) in isolates from homosexual men (35%) than from heterosexual men (49%) or women (49.5%). Sac-4 appears to have an epidemiological association with gonococcal auxotype and serovar rather than a direct association with chlamydial co-infection. PMID- 11860695 TI - A retrospective study of the clinical effectiveness of the treatment of genital co-infection with N. gonorrhoeae and C. trachomatis in Coventry. AB - Concomitant infection of Neisseria gonorrhoeae and Chlamydia trachomatis is frequent and it is common practice to prescribe ancillary treatment for chlamydial infection when infection with N. gonorrhoeae is suspected or confirmed. In Coventry cases are treated as they are diagnosed. Our objective was to determine the clinical effectiveness of treating gonorrhoea and chlamydial infection separately in cases of co-infection. Case notes of co-infection with N. gonorrhoeae and C. trachomatis diagnosed in Coventry GU clinic from March 1989 to February 2000 were reviewed retrospectively. There were 1250 episodes of gonorrhoea, 4127 of chlamydial infections, and both infections were found in 332 cases. The two infections were treated in 322 cases and in 235 cases were treated separately. Ten cases did not come back for treatment of chlamydial infection, which is less than one case per year and 0.2% of total chlamydial infection in 11 years. On the other hand, 918 (73%) of total number of gonorrhoea patients did not have to take unnecessary treatment for chlamydial infection. In some clinical settings co-infection with N. gonorrhoeae and C. trachomatis could be treated separately with significant success and in the long run this might prevent development of antibiotic resistance of C. trachomatis infection. PMID- 11860696 TI - HIV treatments optimism is associated with unprotected anal intercourse with regular and with casual partners among Australian gay and homosexually active men. AB - We examined HIV optimism and unprotected anal intercourse (UAI) in a nationwide sample of gay and homosexually active men. Questionnaires were distributed throughout Australia through pornographic catalogues. 1832 men responded, 1181 (64.5%) classified as gay community attached (GCA) and 651 (35.5%) as non-GCA (NGCA). Mean HIV optimism scores tended toward scepticism rather than optimism, with no significant difference between GCA and NGCA men. Men who had UAI with regular partners were significantly more optimistic than men who did not (P<0.001). There was a significant UAI-casual/GCA interaction (P<0.001). GCA men who engaged in UAI-casual were significantly more optimistic than GCA men who did not (P<0.001). Likewise, NGCA men who had UAI-casual were significantly more optimistic than NGCA men who did not (P<0.001). These findings corroborate earlier evidence that gay men's UAI is associated with - not necessarily caused by - HIV optimism. Importantly, these data come from a broad national sample rather than one drawn from within a 'gay precinct'. PMID- 11860697 TI - High levels of sexual HIV/STD risk behaviour among Roma (Gypsy) men in Bulgaria: patterns and predictors of risk in a representative community sample. AB - Studies on HIV and STD risk factors among vulnerable minority groups in Eastern Europe are underrepresented in the literature. The rapid increase in HIV and STD rates observed throughout the region may quickly affect impoverished, stigmatized, and underprivileged communities. Roma (or Gypsies) constitute such a vulnerable group. A total of 324 men aged 14-37 years were recruited during June July 2001 in a Roma community neighbourhood in Sofia, Bulgaria. HIV/STD risk behaviours were widespread. Men reported a mean of 2.4 female partners in the past 3 months and 77% did not use a condom during their most recent vaginal intercourse. 72% of Roma men said they had engaged in anal intercourse with women in the past 3 months and almost 75% of these heterosexual anal intercourse occurrences were unprotected. 27% reported having sex with other men during their lifetimes, 10% had same-sex anal intercourse partners in the past 3 months, and 58% of the most recent anal intercourse acts by these men were not condom protected. 16% of men reported selling sex, and 32% paid someone for sex. Positive condom-use attitudes, intentions, norms and self-efficacy, as well as younger age and condom availability, were factors associated with lower sexual risk. These factors should be targeted in rapid, comprehensive, and culturally sensitive prevention interventions for Roma communities. PMID- 11860698 TI - HIV risk behaviour among gay and bisexual men in Budapest, Hungary. AB - HIV is a growing public health threat in Central and Eastern Europe. In Hungary and a number of other countries, men who have sex with men (MSM) account for a high proportion of HIV infections. However, there has been very little systematic study of the sexual risk practices and characteristics of MSM in this region. This study surveyed 469 MSM recruited in Budapest gay community venues in June 2001. Half the men (50%) engaged in unprotected anal intercourse (UAI) in the past 3 months. Of these, 40% of men's insertive and 50% of their receptive acts were unprotected, and 25% had multiple AI partners in the past 3 months. 17% of MSM exchanged sex for money, 26% had female partners in the past year, and condoms were used in only 23% of their vaginal intercourse occasions. Multivariate analyses showed that high-risk behaviour was predicted by not having condoms available when needed, weak risk reduction intentions, negative attitudes toward safer sex, being in a steady relationship, and having a bisexual orientation. Community-based HIV prevention programmes focused on the needs of gay or bisexual men in Central and Eastern Europe are urgently needed. PMID- 11860699 TI - STD treatment for men in rural and urban Zimbabwe: choice of practitioner, perceptions of access and quality of care. AB - STD treatment choices and perceptions of treatment services (access, quality of care) by Zimbabwean men are examined in 2 settings: Mbare, a district within the capital city Harare, and Gutu, a rural town. Data collection included a survey of 457 men 18 years of age or older (from a stratified systematic sample), focus groups and key informant interviews. Of 220 cases of self-reported genital symptoms, 81.4% were treated by allopathic practitioners, 9% by traditional/faith healers, 8.6% by the subject, a friend or another person; 1.4% were not treated. Traditional/faith healers were consulted primarily for symptoms involving pain or discomfort rather than ulcers or exudation. Disrespect by the health care provider and consultations that were not private were cited as problems by a small minority of subjects. Significantly more respondents in Mbare than in Gutu had been prevented from obtaining the STD treatment they desired at some point in their life because of cost of treatment (chi(2)=5.23, P=0.02). Given the current deteriorating economic situation in Zimbabwe, cost of treatment may become an even more important impediment in the future. PMID- 11860700 TI - Vulvodynia--new and more effective approaches to therapy. AB - Two cases are described of treatment-resistant vulvodynia that responded well to gabapentin. Gabapentin, an anti-epileptic drug, has been used in the treatment of neuropathic pain such as diabetic neuropathy and post-herpetic neuralgia. However, there has been little experience of its use in the relief of symptoms in vulvodynia and we add our observations to the one report of its use in these circumstances that has been published so far. PMID- 11860701 TI - Recurrent hypersensitivity to Combivir. AB - The non-nucleoside reverse transcriptase inhibitors (NNRTIs) are commonly used in combination antiretroviral therapy and are associated with hypersensitivity reactions on induction therapy. We report a case of recurrent hypersensitivity associated with Combivir, when there was a delay in determining the cause as the NNRTIs were considered to be the more likely cause. PMID- 11860702 TI - Behavioral recovery in a primate model of Parkinson's disease by triple transduction of striatal cells with adeno-associated viral vectors expressing dopamine-synthesizing enzymes. AB - One potential strategy for gene therapy of Parkinson's disease (PD) is the local production of dopamine (DA) in the striatum induced by restoring DA-synthesizing enzymes. In addition to tyrosine hydroxylase (TH) and aromatic-L-amino-acid decarboxylase (AADC), GTP cyclohydrolase I (GCH) is necessary for efficient DA production. Using adeno-associated virus (AAV) vectors, we previously demonstrated that expression of these three enzymes in the striatum resulted in long-term behavioral recovery in rat models of PD. We here extend the preclinical exploration to primate models of PD. Mixtures of three separate AAV vectors expressing TH, AADC, and GCH, respectively, were stereotaxically injected into the unilateral putamen of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-treated monkeys. Coexpression of the enzymes in the unilateral putamen resulted in remarkable improvement in manual dexterity on the contralateral to the AAV-TH/ AADC/-GCH-injected side. Behavioral recovery persisted during the observation period (four monkeys: 48 days, 65 days, 50 days, and >10 months, each). TH immunoreactive (TH-IR), AADC-IR, and GCH-IR cells were present in a large region of the putamen. Microdialysis demonstrated that concentrations of DA in the AAV TH/-AADC/-GCH-injected putamen were increased compared with the control side. Our results show that AAV vectors efficiently introduce DA-synthesizing enzyme genes into the striatum of primates with restoration of motor functions. This triple transduction method may offer a potential therapeutic strategy for PD. PMID- 11860703 TI - Hyaluronidase increases electrogene transfer efficiency in skeletal muscle. AB - Electrogene transfer (EGT) of plasmid DNA into skeletal muscle is a promising strategy for the treatment of muscle disorders and for the systemic secretion of therapeutic proteins. We report here that preinjecting hyaluronidase (HYAse) significantly increases the gene transfer efficiency of muscle EGT. Three constructs encoding mouse erythropoietin (pCMV/mEPO), secreted alkaline phosphatase (pCMV/SeAP), and luciferase (pGGluc) were electroinjected intramuscularly in BALB/c mice and rabbits with and without HYAse pretreatment. Preinjection 1 or 4 hr before EGT increased EPO gene expression by about 5-fold in mice and maintained higher gene expression than plasmid EGT alone. A similar increment in gene expression was observed on pretreatment with HYAse and electroinjection of pCMV/mEPO into rabbit tibialis muscle. The increment of gene expression in rabbits reached 17-fold on injection of plasmid pCMV/SeAP and 24 fold with plasmid pGGluc. Injection of a plasmid encoding beta-galactosidase (pCMV/beta gal/NLS) and subsequent staining with 5-bromo-4-chloro-3-indolyl-beta D-galactopyranoside indicated that HYAse increased the tissue area involved in gene expression. No irreversible tissue damage was observed on histological analysis of treated muscles. HYAse is used in a variety of clinical applications, and thus the combination of HYAse pretreatment and muscle EGT may constitute an efficient gene transfer method to achieve therapeutic levels of gene expression. PMID- 11860705 TI - Interleukin-13 gene therapy reduces inflammation, vascularization, and bony destruction in rat adjuvant-induced arthritis. AB - Rheumatoid arthritis (RA) is a chronic autoimmune inflammatory disease characterized by synovial pannus formation, leukocyte infiltration, and angiogenesis. Adenoviral production of interleukin-13 (IL-13) reduces levels of proinflammatory mediators in an explant model of RA synovial tissue in vitro. To assess this approach in an animal model of arthritis, we compared intra-articular injections of an adenovirus producing rat IL-13 (AxCArIL-13), a control virus, and rat ankles receiving phosphate-buffered saline (PBS) in rat adjuvant-induced arthritis (AIA). We demonstrate that IL-13 levels are normally low in ankles throughout the course of rat AIA. We show that administration of AxCArIL-13 before arthritis onset significantly reduces ankle circumference, paw volume, bony destruction, the number of polymorphonuclear cells (PMNs), the quantity of blood vessels, and levels of monocyte chemoattractant protein (MCP)-1 in ankles. When administered as a treatment to inflamed ankles, AxCArIL-13 decreases articular index scores, paw volumes, bony destruction, vascularization, tumor necrosis factor-alpha (TNF-alpha) levels, and the quantity of monocytes, lymphocytes, and PMNs. Thus, increasing IL-13 levels significantly ameliorates the course of rat AIA, suggesting that similar strategies for the treatment of human RA are worthy of further study. PMID- 11860704 TI - Adenovirus vector-induced inflammation: capsid-dependent induction of the C-C chemokine RANTES requires NF-kappa B. AB - Adenovirus vectors for gene therapy activate responses in the host that result in acute inflammation of transduced tissues. Our previous studies in vivo demonstrate that chemokines, including the C-C chemokine RANTES (regulated on activation, normal T cell expressed and secreted), contribute to the acute inflammation induced by adenovirus vectors. Various first-generation adenovirus vectors, including adCMV beta gal, were equally capable of inducing the expression of RANTES 3 hr after transduction in epithelial HeLa and REC cells. Deletional analysis of the human RANTES promoter revealed that induction by adCMV beta gal required the elements spanning base pairs -90 to -25 of the gene. Electrophoretic mobility shift assays demonstrated that nuclear extracts from adCMV beta gal-transduced HeLa cells bound to an NF-kappa B site at position -54. Overexpression of I-kappa B alpha suppressed adCMV beta gal induction of RANTES, confirming that this process was dependent on the nuclear translocation of NF kappa B. The coxsackievirus-adenovirus receptor (CAR)-independent, serotype 3 adenovirus was equally capable of inducing the expression of RANTES in HeLa cells. This observation suggested that binding to CAR was not specifically required in adenovirus vector-induced RANTES expression. The use of RGD peptides to block adCMV beta gal interactions with alpha(v)-integrins reduced RANTES expression but also transduction efficiency. In CAR-deficient P815 cells, binding of adCMV beta gal to alpha(v)-integrins without efficient cell transduction did not result in increased RANTES expression. Expression of human CAR in P815 cells increased the binding and transduction efficiency of adCMV beta gal and resulted in RANTES expression in these cells. These results suggest that the induction of RANTES by adenovirus vectors is dependent on efficient interaction with its cell surface receptors and vector internalization. Understanding the biology of the host response to adenovirus vectors will impact the design of future generations of these agents aimed at reducing their immunogenicity and improving their safety. PMID- 11860706 TI - Immunoinhibitory DNA vaccine protects against autoimmune diabetes through cDNA encoding a selective CTLA-4 (CD152) ligand. AB - Cytotoxic T lymphocyte antigen 4 (CTLA-4 or CD152) is a strong negative regulator of T cell activity. Like CD28 (a positive regulator) it binds to B7-1 and B7-2, and there is no known natural selective ligand. Monoclonal antibodies to CTLA-4 generally have a masking effect, enhancing rather than suppressing responses. However, a single amino acid substitution in B7-1 (W88 > A; denoted B7-1wa) abrogates binding to CD28 but not to CTLA-4. We constructed plasmids encoding B7 1 or B7-1wa, as cell-surface or Ig fusion proteins. In a bound state, B7-1-Ig enhanced CD3-mediated T cell activation, but B7-1wa-Ig was inhibitory, as expected of a CTLA-4 ligand. To alter immunity in vivo, we inoculated mice intramuscularly (i.m.) with a carcinoembryonic antigen (CEA) plasmid. Gene transfer was amplified by electroporation. Co-injection of a B7-1wa (membrane bound form) plasmid blocked induction of anti-CEA immunity, whereas a B7-1 plasmid was stimulatory. We studied this DNA covaccination method in nonobese diabetic (NOD) mice with autoimmune diabetes. Delivery of either preproinsulin I (PPIns) or B7-1wa cDNA alone did not suppress the autoimmune anti-insulin response of spleen cells. However, co-delivery of B7-1wa and PPIns cDNA abrogated reactivity to insulin and ameliorated disease. Interferon-gamma and interleukin-4 were both depressed, arguing against a Th2 bias. Reactivity to glutamic acid decarboxylase 65, another major islet autoantigen, was not altered and suppressor cells were not identified, suggesting induction of tolerance to insulin by either T cell anergy or deletion. Selective engagement of CTLA-4 through gene transfer represents a novel and powerful way to block autoimmunity specifically. PMID- 11860707 TI - Lentiviral gene transduction of kidney. AB - Gene transfer into kidney holds great potential as a novel therapeutic approach. We have studied the transduction of kidney in vivo after delivery of lentiviral vectors by various routes of administration. A lentiviral vector expressing the bacterial lacZ gene from the cytomegalovirus early promoter was used. The lentiviral vector was delivered into the kidneys of BALB/c mice by retrograde infusion into the ureter, by injection into the renal vein or artery, or by direct injection into the renal parenchyma. Expression of the reporter gene was achieved independently of the route of administration, although it appeared more efficient after parenchymal or ureteral administration. After parenchymal or ureteral infusion, expression of the transgene was localized to the outer medulla and corticomedullary junction. In the case of parenchymal injection, expression of the reporter gene extended to the cortex. Detection of the transgene in the renal proximal tubules was confirmed by in situ polymerase chain reaction after parenchymal or ureteral infusion. On delivery of the lentiviral vector through the renal artery or vein, expression of the reporter gene was markedly lower than was observed with parenchymal or ureteral infusion and was limited to the inner medullary collecting ducts. No apparent histological abnormality was observed after virus administration and transgene expression was stable for at least 3 months. These results provide the first evidence that lentiviral vectors can stably transduce renal cells in vivo and may be effective vehicles for gene delivery to the kidney. PMID- 11860708 TI - Contribution of a combination of ponicidin and acyclovir/ganciclovir to the antitumor efficacy of the herpes simplex virus thymidine kinase gene therapy system. AB - We have previously reported that ponicidin (PND), isolated from Rabdosia ternifolia, potentiates the cell-killing activity of antiherpes prodrugs acyclovir (ACV) and ganciclovir (GCV) in human cancer cells expressing herpes simplex virus thymidine kinase (HSV-TK). To extend these in vitro results to in vivo situations, HSV-TK-expressing HeLa cells were injected into nude mice. The in vivo growth of TK(+) HeLa cells was significantly inhibited by coadministration of PND and ACV, or of PND and GCV, compared with single use of ACV or GCV in spite of lower doses of 1 or 0.25 mg/mouse, respectively. These results indicate that there is a good correlation between this in vivo efficacy and previously reported in vitro efficacy. Because of the insufficiency of incorporation of genes into tumors, bystander cell killing has attracted special interest. In the present study, we determined the ability of PND to potentiate the bystander effects of ACV and GCV in both in vitro and in vivo systems. In vitro combined use of PND with ACV or GCV rendered tumor cells more sensitive to the prodrugs, demonstrating a 1.8- to 97-fold or 2.8- to 26-fold reduction in IC(50) compared with ACV or GCV only, respectively, in 1 to 20% of HSV-TK(+) cells. In the in vivo experiments using nude mice injected with 3 or 10% HSV TK(+) cells, tumor volume was lower in mice treated with a combination of PND and ACV/GCV than in those treated with ACV or GCV only. No toxicity of PND was seen in mice even at a dose 10-fold higher than that used in the in vivo experiments. These novel strategies could provide benefit to ablative cancer gene therapy by making it feasible to use toxic GCV at lower doses and relatively nontoxic ACV. PMID- 11860709 TI - Flow cytometry analysis of adenosine deaminase (ADA) expression: a simple and reliable tool for the assessment of ADA-deficient patients before and after gene therapy. AB - Clinical gene therapy trials for adenosine deaminase (ADA) deficiency have shown limited success of corrective gene transfer into autologous T lymphocytes and CD34(+) cells. In these trials, the levels of gene transduction and expression in hematopoietic cells have been assessed by DNA- or RNA-based assays and measurement of ADA enzyme activity. Although informative, these methods are rarely applied to clonal analysis. The results of these assays therefore provide best estimates of transduction efficiency and gene expression in bulk populations based on the assumption that gene transfer and expression are uniformly distributed among transduced cells. As a useful additional tool for evaluation of ADA gene expression, we have developed a flow cytometry (fluorescence-activated cell sorting, FACS) assay capable of estimating the levels of intracellular ADA on a single-cell basis. We validated this technique with T cell lines and peripheral blood mononuclear cells (PBMCs) from ADA-deficient patients that showed severely reduced levels of ADA expression (ADA-dull) by FACS and Western blot analyses. After retrovirus-mediated ADA gene transfer, these cells showed clearly distinguishable populations exhibiting ADA expression (ADA-bright), thus allowing estimation of transduction efficiency. By mixing ADA-deficient and normal cells and using enzymatic amplification, we determined that our staining procedure could detect as little as 5% ADA-bright cells. This technique, therefore, will be useful to quickly assess the expression of ADA in hematopoietic cells of severe combined immunodeficient patients and represents an important tool for the follow-up of patients treated in clinical gene transfer protocols. PMID- 11860710 TI - A tissue-engineered stent for cell-based vascular gene transfer. AB - Cell-based gene transfer using a stent platform would provide a significant advantage in terms of site-specific gene expression in the vasculature. The current study presents a novel stent design that allows stable in vivo transgene expression over a 4-week period in the vasculature. A mesh-stent coated with fibronectin provided an excellent platform for adherent porcine smooth muscle cells (SMC). Autologous porcine SMC were stably transduced with a plasmid encoding green fluorescence protein (GFP), seeded at high density in the mesh stent, and deployed in the porcine coronary artery. Stable in vivo GFP expression within the mesh-stent (5.2 x 10(5) GFP-positive cells/cm(2) mesh) was demonstrated 1 month after implantation in the porcine coronary artery by fluorescence microscopy and flow cytometry. No significant change in GFP positive cell number within the stent occurred over a 1-month period in vivo when compared to preinsertion. Angiographic and histologic analysis revealed mild neointimal proliferation and no inflammatory infiltrate in the stented segment. This study has implications for treatment of cardiovascular and other diseases where long term cell-based delivery of transgene is a desirable therapeutic option. PMID- 11860711 TI - Gene delivery to pig coronary arteries from stents carrying antibody-tethered adenovirus. AB - Deployment of coronary stents to relieve atherosclerotic obstruction has benefitted millions of patients. However, gene therapy to prevent in-stent restenosis, while promising in experimental studies, remains a challenge. Conventional strategies for viral vector administration utilize catheters that deliver infusions of viral suspensions, which result in suboptimal localization and potentially dangerous distal spread of vector. Stent-based gene delivery may circumvent this problem. We hypothesized that site-specific delivery of adenoviral gene vectors from a stent could be achieved through a mechanism involving anti-viral antibody tethering. Stents were formulated with a collagen coating. Anti-adenoviral monoclonal antibodies were covalently bound to the collagen surface. These antibodies enabled tethering of replication defective adenoviruses through highly specific antigen-antibody affinity. We report for the first time successful stent-based gene delivery using antibody-tethered adenovirus encoding green fluorescent protein (GFP), demonstrating efficient and highly localized gene delivery to arterial smooth muscle cells in both cell culture and pig coronary arteries. Overall arterial wall transduction efficiency in pigs was 5.9 +/- 1.1% of total cells. However, neointimal transduction was more than 17% of total cells in this region. Importantly, when specific antibody was used to tether adenovirus, no distal spread of vector was detectable by PCR, in either distal organs, or in the downstream segments of the stented arteries. Control adenovirus stents, with nonspecific antibody plus adenovirus, demonstrated only a few isolated foci of transduction, and poor site-specific transduction with distal spread of vector. We conclude that a vascular stent is a suitable platform for a localizable viral vector delivery system that also prevents systemic spread of vector. Gene delivery using stent-based anti-viral antibody tethering of vectors should be suitable for a wide array of single or multiple therapeutic gene strategies. PMID- 11860712 TI - Kidney-targeted naked DNA transfer by retrograde renal vein injection in rats. AB - Kidney-targeted gene transfer is expected to revolutionize the treatment of renal diseases. Previous gene transfer methods using nonviral vectors administered via renal arterial, pelvic, or ureteric routes into the glomerulus, tubules, or interstitial fibroblasts have resulted in low-level expression for <1 month. The peritubular capillaries (PTC) network is one of the main targets of kidney transplant rejection and of progressive tubulointerstitial fibrosis, which typifies all progressive renal diseases. To access the PTC, we retrogradely injected a lacZ expression plasmid in Ringer's solution into the renal vein of rats. We detected lacZ expression exclusively in the interstitial fibroblasts near the PTC of the injected kidney by immunoelectron microscopic analysis. Nephrotoxicity attributable to gene transfer was not apparent. We then used a rat erythropoietin (Epo) expression plasmid vector, pCAGGS-Epo, in a reporter assay. We obtained maximal Epo expression when the DNA solution was injected within 5 sec, and with a volume of 1.0 ml. We observed a dose-response relationship between serum Epo levels and the amount of injected DNA up to 100 microg. We detected the transgene-derived Epo mRNA by reverse transcription polymerase chain reaction only in the kidneys injected with pCAGGS-Epo. After an injection of 100 microg of pCAGGS-Epo, the serum Epo levels peaked at 208.3 +/- 71.8 mU/ml at week 5, and gradually decreased to 116.2 +/- 38.7 mU/ml at week 24. A similar pattern was obtained using smaller doses of plasmid, 2 microg or 30 microg of pCAGGS-Epo. Transgene-derived Epo secretion resulted in significant erythropoiesis. This novel technique is simple and safe, allowing high-level and long-term stable gene expression specific to the fibroblasts near the PTC, and should have therapeutic value for future applications in humans. PMID- 11860716 TI - Times change: the public and government. PMID- 11860713 TI - Self-assembly of a virus-mimicking nanostructure system for efficient tumor targeted gene delivery. AB - Molecular therapy, including gene therapy, is a promising strategy for the treatment of human disease. However, delivery of molecular therapeutics efficiently and specifically to the target tissue remains a significant challenge. A human transferrin (Tf)-targeted cationic liposome-DNA complex, Tf lipoplex, has shown high gene transfer efficiency and efficacy with human head and neck cancer in vitro and in vivo (Xu, L., Pirollo, K.F., Tang, W.H., Rait, A., and Chang, E.H. Hum. Gene Ther. 1999;10:2941-2952). Here we explore the structure, size, formation process, and structure-function relationships of Tf lipoplex. We have observed Tf-lipoplex to have a highly compact structure, with a relatively uniform size of 50-90 nm. This nanostructure is novel in that it resembles a virus particle with a dense core enveloped by a membrane coated with Tf molecules spiking the surface. More importantly, compared with unliganded lipoplex, Tf-lipoplex shows enhanced stability, improved in vivo gene transfer efficiency, and long-term efficacy for systemic p53 gene therapy of human prostate cancer when used in combination with conventional radiotherapy. On the basis of our observations, we propose a multistep self-assembly process and Tf facilitated DNA cocondensation model that may provide an explanation for the resultant small size and effectiveness of our nanostructural Tf-lipoplex system. PMID- 11860718 TI - News from the Society for Women's Health Research: subgroup analysis in clinical trials: detecting sex differences. PMID- 11860719 TI - Toward optimal health: the experts discuss physical fitness. Interview by Jodi Godfrey Meisler. PMID- 11860720 TI - Are mammography and palpation sufficient for breast cancer screening? A dissenting opinion. AB - Breast cancer is an equal opportunity killer in that as many as 60%-70% of breast cancer patients have no obvious risk factor(s). Thus, the continued reliance on the importance of risk factors to initiate screening programs may inhibit further inquiry into better diagnostic and prognostic indicators. An extensive review of past and recent literature reveals that mammography is not an objective examination. Its use as a screening tool is facilitated among women 40 years old and older whose breast tissue is primarily fatty and provides better visualization. Younger women are not generally advised to use mammography because of its potentially hazardous effects associated with repeated use of radiation. More importantly, regardless of patient age, radiologists interpret mammograms, and different degrees of interpretation error exist for different radiologists as well as for the same radiologist performing the analysis after a period of time. Thus, the use of mammography as the sole screening tool does not provide patients or physicians with a sense of confidence about sensitivity and specificity. Further, recent enthusiasm to promote mammography screening may give women unrealistic expectations, leading them to falsely believe that a negative examination is assurance that cancer is not present in its earliest detectable stage. We propose to supplement the physical examination and mammography with a third screening modality based on thermal detection monitors. This is a noninvasive and nonradiogenic tool and might enable clinicians to provide patients with every opportunity for early diagnosis. PMID- 11860722 TI - Douching beliefs and practices among black and white women. AB - To ascertain beliefs about douching, douching practices, and their motivational antecedents among adult women living in the southeastern United States, we conducted a telephone survey of a random sample of 535 adult women. Douching was deemed a good hygienic practice by 65% of women, half of whom believed that douching was necessary for good hygiene. These beliefs were more common among black than white women. Older women and less educated women were more likely to believe that douching prevented infections and pregnancies. Physicians were the only discouraging influence regarding douching reported by a substantial proportion of the women. Healthcare providers' advice not to douche is correlated with not douching. Encouragement by mother (OR = 4.7, 95% CI 1.9-11.4), being black (OR = 2.8, 95% CI 1.1-6.9), and having no more than a high school education (OR = 2.2, 95% CI 1.2-4.2) were independently associated with ever (vs. never) douching. A substantial proportion of adult women living in the southeastern United States believe that douching is necessary for good hygiene. Our findings suggest that advice from healthcare providers to discourage the practice may have a salutary effect. PMID- 11860723 TI - A survey of gynecologists concerning menorrhagia: perceptions of bleeding disorders as a possible cause. AB - We sought to determine perceptions and practices of American gynecologists when treating with a woman complaining of menorrhagia, specifically with regard to an underlying bleeding disorder as a potential cause. A mail survey of Georgia members of the American College of Obstetricians and Gynecologists was conducted. The survey response was 52%, and the analysis includes 376 physicians who reported seeing at least one gynecological patient per week. On average, respondents were in practice 20 years and reported that 8% of their patient population complain of menorrhagia. Virtually all physicians reported employing a menstrual history as a starting point for the workup for menorrhagia, and 95% order a hemoglobin/hematocrit determination. About 50% of physicians considered saturating three tampons/pads per 4 hours as excessive, although the criterion varied widely (range 0-24 per 4 hours, SD = 3). The diagnoses considered most likely among reproductive age women were anovulatory bleeding or benign lesions or that the heavy bleeding was within normal limits. Only 4% of physicians would consider von Willebrand disease (VWD) for this age group (women of reproductive age). Among girls near menarche, physicians overwhelmingly consider anovulatory bleeding or bleeding within normal limits the likely diagnoses, and 16% would consider VWD in this age group. Only rarely (3%) do surveyed physicians refer menorrhagia patients to other specialists. Most respondents believe that most menorrhagia is caused by anovulation or is within normal limits. Bleeding disorders are believed to be a rare cause of menorrhagia. PMID- 11860724 TI - Risk factors for exposure to environmental tobacco smoke among ethnically diverse women in the United States. AB - The likelihood of exposure to environmental tobacco smoke (ETS) has been shown to vary across sociodemographic characteristics, health behaviors, and the type of smoking restrictions at work. Women may be particularly at risk. The purpose of our study was to assess differences in the likelihood of exposure to ETS at home and at work among an ethnically diverse sample of women age 40 and older in the United States. We used data from the U.S. Women's Determinants Study and restricted the sample to include only nonsmoking women (n = 2326). Unadjusted and adjusted odds ratios (aOR) for exposure to ETS by sociodemographic characteristics, health risk behaviors, and the type of workplace smoking policy were calculated using logistic regression. Exposure to ETS at home was associated with being American Indian/Alaska Native (aOR 1.5, 95% CI 1.0, 2.6), age 40-44 (aOR 1.6, 95% CI 1.0, 2.6) and 45-54 (aOR 1.8, 95% CI 1.2, 2.6), having eighth grade (aOR 2.1, 95% CI 1.3, 3.6) or high school education (aOR 2.2, 95% CI 1.4, 3.3), inadequate fruit and vegetable consumption (aOR 1.5, 95% CI 1.0, 2.1), and not getting screened for breast cancer (aOR 1.5, 95% CI 1.1, 2.0). Women who did not have regular breast (aOR 1.3, 95% CI 1.9, 1.9) and cervical (aOR 2.0, 95% CI 1.5, 5.3) cancer screening were more likely to be exposed to ETS at work. Exposure to ETS at work was higher among women with some high school education (aOR 2.8, 95% CI 1.5, 5.3) and high school graduates (aOR 3.1, 95% CI 1.9, 5.1) and substantially higher for women who worked where smoking was allowed in some (aOR 15.1, 95% CI 10.2, 22.4) or all (aOR 44.8, 95% CI 19.6, 102.4) work areas. Larger effect sizes were observed for the relationship between selected risk factors and ETS exposure at work than for ETS exposure at home. Among individual risk factors, lower education level was most strongly related to ETS exposure at work. The likelihood of being exposed to ETS at work was highest for women whose workplace smoking policies allowed smoking in some or all work areas. PMID- 11860725 TI - Phytoestrogen and multiple vitamin/mineral effects on bone mineral density in early postmenopausal women: a pilot study. AB - The purpose of the study was to assess the effect of a combination regimen of herbs, vitamins, and minerals on bone mineral density (BMD) in early postmenopausal women via a 2-year, single-blind, uncontrolled, prospective trial. BMD was measured by dual energy x-ray absorptiometry (DEXA) at baseline and at 6, 12, and 24 months. Results of lumbar spine, hip, and forearm densities did not differ significantly from historical controls derived from other recent trials using a similar patient population. Bone mineral losses are reported on an annualized basis over the 2 years for the 12 women who completed the trial: spine (-1.42% per year), hip (-0.43% per year), forearm (-1.42% per year). Six women were withdrawn from the trial by the investigators because of excessive losses of bone mineral, and 1 of these women was diagnosed with hyperparathyroidism. There were no metabolic diseases to explain the losses in the remaining 5 withdrawn subjects. Four of 21 subjects experienced adverse side effects, necessitating dropping out by 3 of these women. In conclusion, the combined treatment regimen of a menopause symptom-oriented herbal blend plus a high potency vitamin/mineral was unsuccessful in protecting women against the predictable acceleration of bone mineral losses associated with early postmenopause. PMID- 11860726 TI - Dried plums improve indices of bone formation in postmenopausal women. AB - Menopause drastically increases the risk of osteoporosis. Aside from drug therapy, lifestyle and nutritional factors play an important role in the maintenance of skeletal health. Our recent findings suggest that dried plums, a rich source of phenolic and flavonoid compounds, are highly effective in modulating bone mass in an ovarian hormone-deficient rat model of osteoporosis. The objective of this study was to examine whether the addition of dried plums to the diets of postmenopausal women positively influences markers of bone turnover. Fifty-eight postmenopausal women not on hormone replacement therapy (HRT) were randomly assigned to consume either 100 g dried plums or 75 g dried apples daily for 3 months. Both dried fruit regimens provided similar amount of calories, fat, carbohydrate, and fiber. Serum and urinary biochemical markers of bone status were assessed before and after treatment. In comparison with corresponding baseline values, only dried plums significantly increased serum levels of insulin like growth factor-I (IGF-I) and bone-specific alkaline phosphatase (BSAP) activity. Higher levels of both serum IGF-I and BSAP are associated with greater rates of bone formation. Serum and urinary markers of bone resorption, however, were not affected by either dietary regimen. The results of this study suggest that dried plums may exert positive effects on bone in postmenopausal women. Longer duration studies are needed to confirm the beneficial effects of dried plum on bone mineral density (BMD) and the skeletal health of postmenopausal women. PMID- 11860727 TI - Usual dietary isoflavone intake, bone mineral density, and bone metabolism in postmenopausal women. AB - Clinical trials of isoflavone supplementation and bone density have been of relatively short duration and yielded inconsistent results. Few studies examined the effects of usual dietary isoflavone intake on bone density, and none examined the effects on markers of bone turnover. This cross-sectional study examines the association of usual, unsupplemented dietary soy intake with bone density at the lumbar spine and hip and markers of bone turnover in postmenopausal women. Participants were 208 postmenopausal Southern California women aged 45-74 years. Information on behavioral and lifestyle factors was obtained, and dietary intake of isoflavones over the past year was assessed with a standardized questionnaire. Bone density was measured at the spine and hip with dual energy x-ray absorptiometry (DEXA). Urinary type I collagen cross-linked N-telopeptides (N-Tx) and pyridinium cross-links (PYR), both markers of bone resorption, and bone alkaline phosphatase (BAP), a marker of bone formation, were assayed. After adjustment for age and obesity, women with the highest daily intake of dietary genistein had N-Tx concentrations 18% lower than those of women who reported no daily genistein consumption (mean 37.29 vs. 45.44, respectively, p = 0.01). After adjustment for all covariates, there were trends toward significant differences in N-Tx (p = 0.09) and spine bone density (p = 0.07), whereby women with the highest level of isoflavone consumption had greater bone density at the spine. These results suggest that usual, unsupplemented dietary isoflavone consumption may be protective against bone loss in postmenopausal women through a reduction in bone resorption. PMID- 11860728 TI - Delay in obtaining conventional healthcare by female internal medicine patients who use herbal therapies. AB - The use of herbal products has been studied in the general population, but few studies have focused on the prevalence of herbal therapy use for treatment of symptoms or disease among female internal medicine patients or on predictors for delaying obtaining conventional care while using herbal therapy. Cross-sectional 34-item self-report surveys were mailed to female patients in two private practice internal medicine sites and interviewer administered to patients in a resident ambulatory clinic. The survey included sociodemographics, medical problems, use of herbal therapies, and whether conventional care was delayed while using herbal therapy. Of 354 patients, 220 (62%) participated. Their mean age was 51 years, and most were Caucasian (77%) and had more than a high school education (60%). Of these, 81 (37%) women used herbal therapies for treatment of symptoms or disease, and use did not differ by study site. Twenty-six (32%) delayed obtaining conventional care while waiting for an herbal product to work, although most eventually obtained conventional care. In multivariate analysis, predictors for delay of care included negative experience with prescription medicines, history of failed treatments, and desire for increased control over personal healthcare. Among female patients of general internists, there was a high prevalence of herbal therapy use for treatment of illness, and some women delay obtaining conventional care while using an herbal product. Predictors for delay may alert physicians to educate their patients before delaying care. PMID- 11860729 TI - Women's health literaturewatch. PMID- 11860730 TI - Viral and immunologic follow up of 4 to 9 years of AIDS treatments by quadruple combinations of virostatics including integrase inhibitors applied in short sequences differing by drug rotation. AB - AIM: To present the 4 to 9 years (median: 6 years) treatment follow up of 10 HIV1 AIDS patients, 9 at AIDS and 1 at A3 stages. METHODS: We have applied from 1992 to 1994, AZT combined with 2 integrase inhibitors, acriflavine and hydroxy-methyl ellipticine. We could shift, in 1994, to combinations of 3 drugs including two more retrotranscriptase inhibitors (RTI), ddI and ddC, and, after 1995, to combinations of 4 drugs including also two other RTI, d4T and 3TC, and 3 protease inhibitors (PI), indinavir, ritonavir, and saquinavir. In 1998, as cobalamine was shown by an in vitro test, to act as integrase inhibitor, vitamin B12 was added in cycles of various lengths. Every three weeks, not only the investigations were repeated, but the virostatics were changed. RESULTS: No grade 2 virostatics toxicity has been registered. The viral loads (VL) decreased according to exponential curves. Their initial parts obeyed first order kinetics. The second parts were and still are asymptotic. The first parts could be rectilinear or sinuous. The sinuosities were associated to cofactors present before treatment (chimerism, UV irradiation, hepatitis C or B and C, brain toxoplasmosis). The asymptotic parts, whose VL were below PCR detectable levels, presented discrete, reversible HIV1 rebounds, associated to other cofactors (such as herpes zoster, herpes 6, CMV, flat condyloma, and influenza). Among immunologic parameters, the monocyte and CTL numbers increased and presented, during the rapidly decreasing part of VL curve, a significant inverse correlation with it. Neither CD4+ nor suppressor T-cell (STC) numbers presented such correlation. Near 100 % of CTL were CD28+. Later, vitamin B12 applications increased monocyte and CD28+ CTL numbers, and appeared to reinforce VL stabilization. CONCLUSION: The combinations of inhibitors affecting 3 retrovirus targets, retrotranscriptase, integrase, and protease have given to 10 out of 10 AIDS patients survivals varying today between 4 to 9 years, in excellent conditions. The UVA-pretreated patient is the only one presenting a not maximally reduced asymptotic VL, while his CD4+ and STC have been absent for 8 years. Other patient VL regressed exponentially to become asymptotic, below PCR detectable levels. PMID- 11860731 TI - Mechanism of growth hormone insensitivity induced by endotoxin. AB - AIM: To investigate the mechanism of growth hormone (GH) insensitivity induced by endotoxin at receptor and post-receptor levels. METHODS: Sprague-Dawley rats were injected endotoxin along with or without GH administration. The liver expression of insulin-like growth factor I (IGF-I), GH receptor (GHR), and suppresor of cytokine signaling (SOCS)-3 mRNA were detected by reverse transcriptase polymerase cha in reaction, the GH levels were measured by radioimmunoassay, the levels of tumor necrosis factor alpha (TNF-alpha) and interleukin-6 (IL-6) were detected by enzyme-linked immunosorbent assay. RESULTS: Serum GH levels had no significant difference compared with control rats after endotoxin injection, however, liver IGF-I mRNA expression was obviously down-regulated in endotoxemic rats. Liver GHR mRNA expression was predominantly down-regulated after LPS injection; although SOCS-3 mRNA was weakly expressed in control rats, it was strongly up-regulated in endotoxemic rats. Endotoxine stimulated the production of TNF-alpha and IL-6, and the elevated IL-6 levels showed a positive correlation with increased SOCS-3 mRNA expression. Exogenous GH could enhance IGF-I mRNA expression in control rats, but it did fail to prevent the decline in IGF-I mRNA expression in endotoxemic rats. Two different LPS dosages (7.5 mg/kg and 5.0 mg/kg) produced the same down-regulation of IGF-I mRNA expression, however, the higher LPS dosage induced more GHR mRNA down-regulation and more SOCS-3 mRNA up regulation. CONCLUSION: The mechanism of growth hormone insensitivity induced by endotoxin was associated with down-regulated GHR mRNA expression at receptor level and up-regulated SOCS-3 mRNA expression at post-receptor level. The inhibition at post-receptor level had close relationship with the increased IL-6 secretion. PMID- 11860732 TI - Opioid mediated anti-nociceptive effect of domperidone and cisapride in mice. AB - AIM: To study the anti-nociceptive effect of domperidone and cisapride in mice. METHODS: Initially, the effect of these drugs on motor activity was tested using rotarod. The anti-nociception was tested using chemical and mechanical assay. In the chemical assay, the number of abdominal constrictions either in the saline treated animals or in the domperidone/cisapride (1, 5, or 10 mg/kg either po or ip) treated mice, were recorded for a period of 30 min after acetic acid challenge (10 mL/kg, of 0.6 % acetic acid ip). In the tail clip assay, the time taken by the mouse to make attempts to dislodge the bulldog clamp placed at the tail (reaction time) was recorded with a cut off time of 30 s. The role of opioid pathways was examined by pretreating the animals with naloxone (1 mg/kg, ip) 30 min prior to domperidone and cisapride. RESULTS: Domperidone and cisapride, both reduced the number of abdominal constrictions when given orally or intraperitoneally. Domperidone (5 mg/kg) inhibited it to the extent of 57.0 % after po and 54.6 % after ip. The inhibition after cisapride (5 mg/kg) was 65.1 % (po) and 71.6 % (ip). Naloxone pretreatment reduced this inhibition (57.0 % vs 10.3 % for domperidone and induced hyperalgesia by antagonizing the inhibition and enhanced analgesia to the extent of 28.4 % for cisapride). The reaction time was increased by domperidone (10 mg/kg, ip) from 1.6 s +/- 1.0 s to 14.8 s +/- 0.5 s and cisapride (10 mg/kg, ip) from 3.3 s +/- 1.0 s to 14.8 s +/- 0.5 s. CONCLUSION: Domperidone and cisapride exhibited a significant anti-nociceptive activity after oral as well as intraperitoneal administration. A role for opioid pathways is indicated. Since domperidone is likely to exert less extrapyramidal effects, it can be substituted for metoclopramide, which is now widely used as an analgesic either alone or as an adjuvant. PMID- 11860733 TI - Assessing physiological concentrations of endogenous substances in situ by inducing calcium oscillations in vitro. Case of liver. AB - AIM: To identify the physiological concentration ranges of norepinephrine (NE), vasopressin (VP), and ATP in the rat liver. METHODS: Rat hepatocytes were isolated by collagenase perfusion. Isolated cells were loaded with the fluorescent calcium indicator Fura-2 acetoxymethyl ester (Fura-2 AM). The effects of different concentrations of norepinephrine, vasopressin, and ATP on intracellular calcium concentration ([Ca2+]i) increases in the freshly isolated rat hepatocytes were investigated. [Ca2+]i was measured by microfluorometry and recorded as fluorescence ratios (F340/F380). RESULTS: NE, VP, and ATP induced increases in [Ca2+]i in a concentration-dependent manner. At lower concentrations, [Ca2+]i tended to show an oscillatory increase; with increasing concentrations, [Ca2+]i in more cells tended to show phasic or plateau increases. NE, VP, and ATP concentrations likely to induce an oscillatory [Ca2+]i response were 100 - 500 nmol/L, 50 - 100 pmol/L, and < 1 micromol/L respectively. CONCLUSION: Physiological concentrations of NE, VP, and ATP are 100 - 500 nmol/L, 50 - 100 pmol/L, and < 1 micromol/L respectively in the rat liver. PMID- 11860734 TI - Involvement of heme oxygenase-1 in delayed cardioprotection induced by monophosphoryl lipid A in rats. AB - AIM: To explore whether the heme oxygenase-1 (HO-1) pathway is involved in the delayed cardioprotection induced by monophosphoryl lipid A (MLA). METHODS: Sprague-Dawley rats were pretreated with MLA 24 h before the experiment. Ischemia reperfusion injury was induced by 60 min coronary artery occlusion followed by 3 h reperfusion. Infarct size, the serum creatine kinase (CK) activity, the serum content of nitric oxide (NO), and expression of HO-1 mRNA and protein in the heart were measured. RESULTS: Pretreatment with MLA (500 microg/kg, ip) markedly reduced infarct size and CK release and increased the serum content of NO (P < 0.01). The effects of MLA were completely abolished by pretreatment with L nitroarginine methyl ester (L-NAME 10 mg/kg, ip), an inhibitor of NO synthase (P < 0.01), or Zinc protoporphyrin IX (45 micromol/kg, ip), an inhibitor of HO (P < 0.01). MLA caused a significant increase in the expression of HO-1 mRNA and protein, an effect which was not affected by L-NAME (P > 0.05). CONCLUSION: The results suggest that the HO-1/NO pathway is involved in the delayed cardioprotection induced by MLA. PMID- 11860735 TI - Endomorphins inhibit contractile responses of rat thoracic aorta rings induced by phenylephrine and angiotensin II in vitro. AB - AIM: To study the effects of opioid receptor agonists endomorphin-1 and -2 on contractile responses of rat thoracic aorta rings to phenylephrine (PE) and angiotensin II (Ang II), and their possible mechanism in vitro. METHODS: Isometric tension recording was progressed in thoracic aorta rings from Wistar rats. RESULTS: Pretreatment of morphine, endomorphin-1 and -2 (0.1, 1, and 10 micromol/L) could inhibit the contractile responses of the endothelium-intact aorta rings to PE (0.1 micromol/L) and Ang II ( 1 micromol/L) in a concentration dependent manner (P < 0.01), but could not inhibit the contraction of rings without endothelium (P > 0.05). Naloxone (1 micromol/L) could partially antagonize the effects of endomorphine-1 and -2 (P < 0.01). N(omega)-nitro-L arginine (L-NNA, 10 micromol/L) or endothelial rubbing could completely blocked the effects of morphine, endomorphine-1 and -2 (P < 0.01). CONCLUSION: Endomorphin-1 and -2 could inhibit PE- and Ang II-induced contractions of rat aorta rings, which was partially by naloxone-sensitive mechanism and related to the release of nitric oxide from vascular endothelium. PMID- 11860736 TI - VEGF protects bovine aortic endothelial cells from TNF-alpha- and H2O2-induced apoptosis via co-modulatory effects on p38-and p42/p44-CCDPK signaling. AB - AIM: To investigate the effect of VEGF on TNF-alpha- or H2O2-induced apoptosis in cultured bovine aortic endothelial cells (BAEC) and the underlied signal transduction mechanisms related to Ca2+-calmodulin dependent protein kinase (CCDPK). METHODS: BAEC were cultured and passaged in DMEM. Morphologic changes and quantification of apoptotic cells were determined under fluorescence microscope with Hoechst 33258 staining. Cell viability was detected with MTT method. DNA fragmentation was visualized by agarose gel electrophoresis. The expression of phospho-p38 and phospho-p42/p44 CCDPK was measured by Western blotting. RESULTS: TNF-alpha 5000 kU/L and H2O2 300 micromol/L elicited DNA fragmentation in BAEC. Vascular endothelial growth factor (VEGF) 100 microg/L significantly protected BAEC from apoptosis induced by TNF-alpha or H2O2, as shown in cell viability assay and apoptotic cell counting. DNA fragmentation induced by TNF-alpha or H2O2 was also reduced by VEGF 100 microg/L. VEGF enhanced TNF-alpha and H2O2 stimulated expression of phospho-p42/p44 CCDPK, simultaneously inhibited TNF-alpha- and H2O2-induced activation of phospho-p38 CCDPK. Both the VEGF-induced up-regulation of phospho-p42/p44 CCDPK and its anti-apoptotic action were prevented by the specific p42/p44 CCDPK inhibitor U0126. CONCLUSION: VEGF protects BAEC from apoptosis induced by TNF-alpha and H2O2, and its co-modulatory effects by activation of p42/p44 CCDPK signaling together with inhibition of p38 CCDPK signaling appear to be an important mechanism for its survival effect on endothelial cells. PMID- 11860737 TI - Effects of ropivacaine on sodium, calcium, and potassium currents in guinea pig ventricular myocytes. AB - AIM: To study the effects of ropivacaine (Rop) on sodium current (INa), L-type calcium current (ICa-L), inward rectifier potassium current (IK1), and delayed rectifier potassium current (IK) in isolated guinea pig ventricular myocytes. METHODS: Whole cell patch-clamp techniques were used in our experiment. RESULTS: At potential of -40 mV, Rop 10, 50, and 100 micromol/L decreased sodium current by 8.3 %, 33.3 %, and 62.5 %, respectively and prolonged the time constant of INa inactivation by 8.2 %, 24.7 %, and 64.4 %, respectively (n = 5 cells from 3 animals, P < 0.05). At potential of +10 mV, Rop 50 and 100 micromol/L decreased L type calcium current by 7.6 % and 22.5 %, and prolonged the slow time constant of ICa-L inactivation by 15.5 % and 33.0 %, respectively (n = 5 cells from 4 animals, P < 0.05). Rop 50 and 100 micromol/L did not markedly change the peak current of delayed rectifier potassium current and inward rectifier potassium current (n = 5 cells from 3 animals, P > 0.05), respectively. CONCLUSION: Rop depressed INa and ICa-L, which may be related to its cardiotoxic effect PMID- 11860738 TI - Adeno-associated virus mediated expression of human erythropoietin in vitro. AB - AIM: To investigate the expression of human erythropoietin (hEpo) in 293 cell line using adeno-associated virus (AAV) vector, and provide information for its potential application in human gene therapy. METHODS: The human Epo cDNA was inserted into the AAV cassettes, and recombinant adeno-associated virus (rAAV EPO) vectors were generated by cotransfection methods without helper virus. The expression of hEpo was detected by RT-PCR and ELISA methods. RESULTS: A recombinant adeno-associated virus expressing hEpo was constructed and generated successfully. Positive signal of mRNA was detected from 293 cells transduced with rAAV-EPO, and the hEpo level in supernatant reached 500 U/L. CONCLUSION: rAAV-EPO expressed biologically active hEpo effectively, suggesting that it is a suitable gene expression tool for further study in vivo experiments. PMID- 11860739 TI - Norepinephrine metabolism in neuron: dissociation between 3,4 dihydroxyphenylglycol and 3,4-dihydroxymandelic acid pathways. AB - AIM: To investigate the pre-synaptic metabolism of norepinephrine (NE), judged by variations in plasma concentration of 3,4-dihydroxyphenylglycol (DHPG) and 3,4 dihydroxymandelic acid (DOMA). METHODS: Pithed and electrically stimulated (2.5 Hz) rats were given intravenous infusion of exogenous NE (6 nmol . kg-1 . min-1). Plasma NE, DHPG, DOMA, and the activities of mono- amine oxidases (MAO) were measured with the radio-enzymatic assay. RESULTS: Exogenous NE induces an about 100-fold increase in plasma NE concentration while blood pressure remained within normal limits. A 12-fold increase in plasma DHPG and 1.2-fold increase for DOMA were observed. When NE transportation across the pre-synaptic membrane was inhibited by desipramine (2 mg/kg, iv), a great reduction in plasma DHPG concentration (about 25 % of control) was observed while DOMA remained unchanged. When MAO-A activity was inhibited to 25 % of control by clorgyline (2 mg/kg, iv) and MAO-B to 30 % by deprenyl, the plasma DHPG and DOMA concentrations were reduced to 15 % and 70 % of controls, and to 26 % and 76 % of controls, respectively. When clorgyline and deprenyl were combined, plasma DHPG was vanished (less than 2 % of control) while plasma DOMA remained in the same range (72 % of control). CONCLUSION: The metabolizing system of NE in pre-synapse, associating with the pre-synaptic reuptake plus oxidative deamination on the external membrane of mitochondria, is predominant for the reduction to DHPG. PMID- 11860740 TI - Effects of brain histamine on memory deficit induced by nucleus basalis-lesion in rats. AB - AIM: To investigate whether or not brain histamine was involved in memory deficits induced by lesions of nucleus basalis magnocellularis (NBM) in rats. METHODS: Passive avoidance response was used to measure memory process in rats, and NBM was bilaterally lesioned by injection of ibotenic acid (6 microg/site). RESULTS: Icv injection of histamine (500 ng), and ip injection of histidine (1500 mg/kg), metoprine (10 mg/kg) or tacrine (3, 5 mg/kg) ameliorated memory impairment induced by NBM lesion regarding passive avoidance response. The ameliorating effect of histidine was antagonized by pyrilamine (2-5 mg/kg), a H1 antagonist, but not by zolantidine, a H2-antagonist. CONCLUSION: Histaminergic neurons play an important role in learning and memory via H1-receptor, and its action may be due to cholinergic neurons. PMID- 11860741 TI - Protective effects of melatonin on cortico-hippocampal neurotoxicity induced by amyloid beta-peptide 25-35. AB - AIM: To study the effects of melatonin on primary rat cortico-hippocampal neurotoxicity induced by amyloid beta-peptide 25-35. METHODS: The neuronal morphology was observed by phase-contrast microscopy. The neurotoxicity was quantitatively estimated by measuring lactate dehydrogenase (LDH) released into the culture medium from the damaged neurons. The neuronal metabolic state was quantified by the reduction of 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyltetrazolium bromide (MTT). RESULTS: Treatment of primary rat cortico hippocampal neurons with amyloid beta-peptide 25-35 (20 micromol/L) for 24 h caused a significant decrease in neurocyte viability (P < 0.01, compared with control). Melatonin (1 or 10 micromol/L) reduced the neurotoxicity induced by amyloid beta-peptide 25-35. CONCLUSION: Amyloid beta-peptide 25-35 could exert direct cytotoxicity on rat cortico-hippocampal neurocytes and melatonin concentration-dependently rescued cultured neurons from exposure to amyloid beta peptide 25-35 induced injury. PMID- 11860742 TI - Identification of three sulfate-conjugated metabolites of berberine chloride in healthy volunteers' urine after oral administration. AB - AIM: To identify the structure of unknown metabolites of berberine (Ber) in human urine after oral administration. METHODS: Urine samples were obtained from 5 volunteers after they orally took Ber chloride 0.9 g per day for three days. Metabolites in urine samples were isolated and purified by polyporous resin column chromatography. The individual metabolites were identified mainly using electrospray ionization mass spectroscopy (ESI-MS) and proton nuclear magnetic resonance (1H NMR) spectroscopy. RESULTS: Three unknown metabolites (M1, M2, and M3) were isolated. They were susceptible to arylsufatase. ESI-MS measurements of M1, M2, and M3 produced quasimolecular ions [M+H]+, m/z 17.9, 404.0, and 402.0 respectively. Especially, each of them produced a characteristic protonated ion [M-80+H]+, which can be ascribed as quasimolecular ions lost a SO3 fragment. 1H NMR spectra of the metabolites were also obtained and each of 1H signals was assigned. CONCLUSION: Structures of M1, M2, and M3 were firmly identified as jatrorrhizine-3-sulfate, demethyleneberberine-2-sulfate, and thalifendine-10 sulfate, and the major metabolite was M2. PMID- 11860743 TI - Stereoselectivity in renal clearance of trans-tramadol and its active metabolite, trans-O-demethyltramadol. AB - AIM: To study the stereoselectivity in renal clearance of trans-tramadol and its active metabolite, trans-O-demethyltramadol. METHODS: The right kidneys were isolated from male SD rats and perfused with 100 mL of perfusate medium containing trans-tramadol 300 microg/L or trans-O-demethyltramadol 50 microg/L. After perfusion, the concentrations of the enantiomers of trans-tramadol and trans-O-demethyltramadol in the perfusate and urine were determined by high performance capillary electrophoresis. The enantiomeric ratios were calculated. RESULTS: After the kidneys being perfused with trans-tramadol hydrochloride, the concentration of (+)-trans-tramadol was higher than that of (-)-trans-tramadol, and the concentration of (+)-trans-O-demethyltramadol was lower than that of (-) trans-O-demethyltramadol in the perfusate; meanwhile, (+)-trans-tramadol was more than (-)-trans-tramadol, and (+)-trans-O-demethyltramadol was less than (-)-trans O-demethyltramadol in the urine. After the kidneys being perfused with trans-O demethyltramadol, the concentration of (+)-trans- O-demethyltramadol was lower than that of (-)-trans-O-demethyltramadol in the perfusate, and (+)-trans-O demethyltramadol was more than (-)-trans-O-demethyltramadol in the urine. CONCLUSION: The renal clearance of trans-tramadol was stereoselective. The O demethylation of trans-tramadol was stereoselective in the kidneys, (-)-trans tramadol being preferentially metabolized. The renal clearance of trans-O demethyltramadol was also stereoselective, the (+)-enantiomer being preferentially cleared into the urine. PMID- 11860744 TI - Restricted-access media high pressure liquid chromatography vs fluorescence polarization immunoassay for analysis of carbamazepine in human plasma. AB - AIM: To compare restricted-access media high performance liquid chromatographic (RAM-HPLC) method with fluorescence polarization immunoassay (FPIA) for analysis of carbamazepine (CBZ) in human blood. METHODS: An RAM-HPLC method was established for the determination of CBZ in plasma. RESULTS: The two methods do not need sample clean-up prior to analysis and they have almost 100 % recovery and good reproducibility. There is a good correlation between the CBZ concentration in venous plasma samples determined by FPIA and that in both venous and fingertip plasma samples obtained by RAM-HPLC, the correlation coefficients being 0.989 and 0.995, respectively. It is shown by t-test that the data sets of venous and fingertip plasma samples given by RAM-HPLC are consistent with each other but significantly different from the results obtained by FPIA. CONCLUSION: Both direct injection RAM-HPLC and FPIA may be applied in determining CBZ in therapeutic drug monitoring (TDM). FPIA is well-suited to the routine TDM. RAM HPLC is more useful in TDM related research and especial cases. PMID- 11860745 TI - Inhibition of 11beta-hydroxysteroid dehydrogenase in guinea pig kidney by three bioflavonoids and their interactions with gossypol. AB - AIM: To study the effects of some bioflavonoids on the gossypol-induced hypokalemia. METHODS: The 11beta-hydroxysteroid dehydrogenase (11beta-OHSD) protein was prepared from guinea pig kidney. The activity of 11beta-OHSD with NAD as the coenzyme was measured by HPLC. The drug interaction was analysed by isobolographic method. RESULTS: The 11beta-OHSD can be inhibited by some bioflavonoids. The IC50 (95 % confidence limits) values were: quercetin 164 (79 - 341) micromol/L, morin 913 (385 - 2173) micromol/L, and naringenin 2193 (1114 - 4315) micromol/L. When the 11beta-OHSD was treated with quercetin, tangeretin, morin, naringenin plus gossypol, the combination index (CI) values were 0.92, 0.85, 0.98, and 1.01 respectively. CONCLUSION: The interaction of some bioflavonoids with gossypol might be one of the factors for gossypol-induced hypokalemia. PMID- 11860746 TI - [National sentinel surveillance of HIV infection in China from 1995 to 1998]. AB - OBJECTIVE: To monitor the trend of HIV infection in China in order to provide accurate information for AIDS prevention policymakers and planners. METHODS: Since 1995, cross - sectional sentinel surveys have repeatedly been conducted in China. The target populations in these surveys were STD clinic attenders, prostitutes, drug users, long distance truck drivers, pregnant women, and blood donors. RESULTS: In 1995, survey results found no HIV infection among drug users in 8 sentinel surveillance sites. In 1998 however, HIV infection had been found in 17 of 19 sentinel surveillance sites. HIV infection prevalence rates among drug users in Urumqi had increased from 0% in 1995 to 28.8% in 1998 and in Guangxi and Guangdong, from 1% in 1997 to 12.8% and 10.4% respectively. The highest HIV prevalence rate was 82.2% among drug users in Yining city. The HIV prevalence rate remains low in other risk groups but has increased among prostitutes, whose rate of insistent condom use remains low, and among STD clinic attenders. Needle and syringe sharing among drug users have also increased in many sites. CONCLUSION: HIV infection is spreading rapidly among drug users. There are many risks involved in the spread of HIV among prostitutes and their clients, and thus more efficient interventions are urgently needed. PMID- 11860747 TI - [The survey of prevalence of HIV infection among paid blood donors in one county in China]. AB - OBJECTIVE: To estimate the prevalence rate of HIV infection among paid blood donors and to analyse high risk factors. METHOD: All residents between 18 to 60 years old were selected from on county in March 1999. RESULTS: There were 210 paid blood donors among 730 interviewed subjects. The HIV prevalence rate among paid blood donors was higher (9.1% or 19/210) than that of others (0.6% or 3/520). The HIV prevalence rate was higher in plasma donors (25.9%) than that (2.6%) among 210 blood donors. Donors who donated blood/plasma more than 10 times each year had a higher prevalence rate (13.5%) than those who donated less than 10 times each year (2.8%). Donors who had donated blood but stopped donation before 1995 had a higher prevalence rate (18.3%) than those who started donation after 1996 (1.4%). The results of KAP survey showed that blood donors had a low knowledge about AIDS. CONCLUSION: There was a higher prevalence rate of HIV infection among paid blood donors in some regions. It should take measures to prevent further transmission HIV through paid blood donors. PMID- 11860748 TI - [Residual risk research of HIV infection after blood screening in one county in China]. AB - OBJECTIVE: To estimate the risk of HIV infections from blood transfusions, to evaluate existing blood screening measures, and to make recommendations to improve current blood screening practices. METHODS: Research was conducted in a county blood station. A statistical model and retrospective method were used to analyse the data. RESULTS: Between June 1995 and March 1998, 19 blood donors tested positive for HIV antibodies and among them, 7 were repeat blood donors. Twenty - two blood recipients were identified from these 7 donors during six months, 9 are still alive today, while one patient was infected with HIV from a blood transfusion while undergoing an operation. The residual risk of the blood station using a statistical model and retrospective methods were 1/1,773 - 1/28,867 and 1/2,008 - 1/2,730 donations respectively. CONCLUSION: The residual risk of HIV after screening in the county blood station was high. We recommend further improvements to current blood screening practices. PMID- 11860749 TI - [The survey of HIV prevalence among drug users in Guangxi, China]. AB - OBJECTIVE: To understand the distribution and related risk factors of HIV infection among drug users in Guangxi. METHOD: Three hundred and twenty five drug users were tested for serum HIV antibody, with enzyme linked immunosorbent assay. RESULTS: One hundred and sixteen drug users (35.7%) were found to be HIV positive. The main risk factors associated with HIV infection were intravenous drug users and needle sharing among drug users. The HIV prevalence was 40.9% (114/279) among IDUs, and only 4.3% (2/46) among oral drug users. The phenomenon of sharing injecting equipment (76.7%) among intravenous drug users in Guangxi represents a very serious problem. CONCLUSION: There is a high risk of HIV infection among drug users in Guangxi. Prevention programs urgently need to be implemented. PMID- 11860750 TI - [The natural history of HIV infection among IDUs in Ruili, Yunnan province, China]. AB - OBJECTIVE: To study the natural history of HIV infections among intravenous drug users. METHODS: Six hundred and twenty five HIV infected people in Ruili, Yunnan province were observed. Retrospective data was analysed combined with a cross - sectional survey. RESULTS: The number of death was 190 among 625 HIV - positive people. The cumulative death rate among HIV infected persons was 76.9 per 1,000 person - year, and the AIDS related death rate was 19.8 per 1,000 person - year. The median survival period of HIV infected persons was 9 years, and the median incubation period was about 8 years. CONCLUSION: Of HIV infected people in Ruili, the median survival and incubation period were shorter compared with those in the United States and Western Europe. PMID- 11860751 TI - [A study of simian virus 40 infection and its origin in human brain tumors]. AB - OBJECTIVE: To study the status of simian virus 40 (SV40) infection and its origin in human brain tumors. METHODS: Polymerase chain reaction (PCR) and Dot blot hybridization were used to detect SV40 DNA sequences in 516 human brain tumor tissues, 80 peripheral blood cells and 50 sperm fluids from healthy individuals, 100 human embryo tissues from artificial abortion, 30 normal brain tissues and two human glioma cell lines: SHG44 and BT325. RESULTS: SV40 DNA sequences were found in 36.4% of human brain tumors (188/516), 16.3% of healthy peripheral blood cells (13/80), 22.0% of healthy semen (11/50), 8.0% of human embryo tissues (8/100) and 6.7% of normal brain tissues (2/30). SV40 DNA sequences were also detected in SHG44 and BT325 cell lines. The positive rates of SV40 DNA in human brain tumors, peripheral blood cells and semen from healthy donors were significantly higher than those in human normal brain and embryo tissues (P < 0.05). CONCLUSIONS: (1) SV40 has a higher infection rate in human brain tumors, (2) SV40 is closely related to the etiopathogenesis of human brain tumors, (3) the ways of SV40 spread may due to both horizontal and diaplacental infections in human population. PMID- 11860752 TI - [A case control study on the distribution of apolipoprotein AI gene polymorphisms in the survivors of atherosclerosis cerebral infarction]. AB - OBJECTIVE: To investigate the frequency of the restrict fragment length polimorphisms (RFLP) at 2 MspI sites in the 5' - end of apolipoprotein AI (ApoAI) gene and its potential interaction to atherosclerotic cerebral infarction (ACI) in Chinese Han population. METHOD: Polymerase chain reaction (PCR) technique was used in a sample of 199 cases with documented ACI and 204 healthy matched by age and sex individuals selected from Chinese Han nationality in Beijing. The studied loci include promoter region (-75 bp) and the intron - 1 (+83 bp). RESULTS: (1) Both ACI and control groups M1 + + and M1 + - were major frequent genotypes in Beijng area, M2 + + were major frequent genotypes in intron - 1; (2) M1 + - genotype appeared more common among patients than in controls (0.487 vs 0.368, P < 0.05, OR = 1.64); (3) Individuals with M1 + -/M2 + + haplotype were significantly increased in patients compared with controls (0.437 vs 0.314, P < 0.05, OR = 1.70). CONCLUSION: It is suggested that both M1 + - genotype and M1 + /M2 + + haplotype of ApoAI gene might associate with ACI in our study. PMID- 11860754 TI - [A study on the epidemiology and preventive measure on epidemic haemorrhage fever in army barrack area in Shandong]. AB - OBJECTIVE: In order to guide the army and civilians to carry out the prevention on epidemic haemorrhage fever (EHF), it's necessary to elucidate the epidemic feature of natural epidemic foci and the risk factors of EHF in the army - stationed areas. METHODS: Samples were tested by pathogenic and serologic means before data was collected and analyzed by case - control or family - control studies. RESULTS: Cases belong to house - mouse type (91.48%), field - mouse type (7.21%) and unclassified (1.31%) coexist in the stationed area. In the two types of noticed cases, no significant difference in distributions of sex, profession and ages. Most house - mouse type cases occurred in the peak time from March to June and field - mouse type in November to next February. The type of EHF virus antigen of mice and the type of serum were both accordant with their host types. Dwelling in border area, living rooms with mice, dogs raising, injury of mouth tissue, eating food contaminated by mice, and household with mice living around were more frequently seen in cases themselves and in families of the cases than in controls of both groups (P < 0.05). The mice intensity could be under control to 1.0%, if comprehensive measure on mice control can be implemented. CONCLUSIONS: The stationed area is an epidemic area mixed with both house - mouse and field - mouse types, with majority of house - mouse type. It was house - mouse type virus infection that caused the epidemic area expanded and the number of cases increased radically these years. EHF was epidemic in spring and summer season in the stationed area mainly because the infections were among family members. Food contaminated by mice or close contact with mice played an important role. Mice eradication and prevention were the leading measures to prevent EHF, from our more than ten years of experiences. PMID- 11860753 TI - [An epidemiologic study of irritable bowel syndrome in Beijing - a stratified randomized study by clustering sampling]. AB - OBJECTIVE: To explore the prevalence of irritable bowel syndrome (IBS) and its risk factors in Beijing. METHODS: Phase I: a screening program for IBS was carried out in Beijing area according to symptoms using both Manning (modified including constipation) and Rome criteria. Random clustered sampling involving 2 486 subjects of the inhabitants was carried out under stratification of city, suburban and rural areas. Sample size of each studied area was in proportion to the population of the area. The range of age was 18 - 70. All subjects studied were requested to fill in a questionnaire assisted by trained doctors or medical students during the visit to their families. Phase II: a small number of patients who fulfilled the Manning criteria were further selected to undergo detail clinical examination in the hospital including laboratory examination, abdominal ultrasonography, colonoscopy or/and barium enema to exclude organic disease of the colon. Prevalence of IBS of the population was then adjusted by the rate of correct diagnosis during phase II study. Minnesota Multi - Personality Indices was used in some cases. Potential risk factors were explored by comparing frequencies among IBS group and non - IBS group using chi(2) and logistic analysis of multifactors. RESULTS: The adjusted point prevalence of IBS in Beijing according to Manning criteria was 7.26%, in contrast to Rome criteria - 0.82%. Higher prevalence rate of IBS in the city (10.50%) than in rural areas (6.14%) (P < 0.001) was noticed. Male to female ratio was 1:1.15 with majority of IBS fell in age 18 - 40 (51.6%), and among intellectuals. Our study indicated that history of dysentery (OR = 3.00), exposure to coldness (OR = 1.55), ingestion of cold food and raw materials (OR = 1.24) might serve as the most important risk factors (P < 0.001). Patients with IBS seemed to have a higher tendency of abnormal personalities. CONCLUSION: IBS is a common disorder in Beijing which deserves greater care. PMID- 11860755 TI - [Studies on three types of natural foci of tsutsugamushi disease in eastern part of China]. AB - OBJECTIVE: In order to identify the types of natural foci of tsutsugamushi disease to provide basis for prevention, a series of studies were carried out in Fujian, Zhejiang and Jiangsu Provinces. METHODS: (1) Representative points for investigation in three Provinces were selected. (2) Data on geographical landscape and epidemiology were collected. (3) Field survey and experimental studies on rats and chigger mites in relation to the disease. RESULTS: Three representative types of natural foci of tsutsugamushi disease were demonstrated. They were (1) Coastal island type, mainly in Fujian with major reservoir host as Rattus losea and major vector mite as L. deliense; prevalent season: summer; type of Rt: Gilliam: (2) Inland mountain - forest type in Zhejiang with major reservoir host as R. confucianus and major vector miteas L. gaohuense; prevalent season: summer; type of Rt: Gilliam. (3) Inland plain - hills type in Jiangsu with major reservoir hosts: Apodemus agrarius, R. confucianus, R. norvegicus and Crocidura lasiura; major vector mite: L. scutellare; prevalent seasons: autumn and winter; type of Rt: Kawasaki. CONCLUSION: The types of Rt, reservoir hosts, vector chigger mites and epidemiological features of tsutsugamushi disease in Fujian, Zhejiang and Jiangsu Province were found different. Thus, the preventive measures should also be distinct from one another. PMID- 11860756 TI - [A case - control study on risk factors for hip fracture in the middle - aged and elderly in Beijing]. AB - OBJECTIVE: To investigate the risk factors for hip fracture in women and men aged 50 years and older. METHODS: A matched case - control study was carried out in the urban area of Beijing from Jan. 1994 to May 1996. A total number of 201 cases having had hip fracture were identified by X - ray films in four hospitals of Beijing. Four hundred and two neighbourhood controls were individually matched with these cases by age and sex. Data were analyzed by statistical technique of Chi-square test and conditional logistic regression. RESULTS: In a multiple conditional logistic regression model, low dietary calcium intake, previous history of more sedantary position, lack of calcium supplement, low body mass index, current low physical exercise, never living in rural, sedentary position at work, previous fractures of any type after the age of 50, history of family hip fracture and diabetes mellitus were significantly associated with increased risk of hip fracture. In addition, hip fracture was related to the age of menopause and duration of breastfeeding in women, as well as body height in men. CONCLUSION: Hip fracture is a disease with multiple risk factors which are different with sex. PMID- 11860758 TI - [A case - control study of gallstone disease in female population in Taicang]. AB - OBJECTIVE: To investigate the risk factors of gallstone disease in female population in Taicang. METHODS: A case - control study conducted on 144 cases and 371 controls aged 35 - 79 years in five villages. Diagnosis of gallstone disease was assessed by real - time ultrasonography. The case was considered as presence of gallstones at echograph and previous cholecytectomy. The controls were selected from a random sample of female participates. RESULTS: Through univariate and multivariate analysis, gallstone disease was significantly associated with age (OR = 1.80), family history of gallstone disease (OR = 3.66), hypertension (OR = 1.93), fat intake (OR = 1.77), number of pregnancies (OR = 1.44) and menopause (OR = 1.67). Drinking water from deep well (OR = 0.54) was inversely associated with gallstone disease. CONCLUSION: The main risk factors of female gallstone disease were age, family history of gallstone disease, fat intake, number of pregnancy and menopause. Drinking water from deep well was a protective factor of gallstone disease. PMID- 11860757 TI - [A study on general status and affecting factors on infant intellectual development in Changsha city]. AB - OBJECTIVE: To evaluate the intellectual level of infants in Changsha city and analyze the main factors that affect infant intellectual development. METHODS: Two hundred and sixty infants were measured by Fagan Test of Infant Intelligence, who were randomly selected from Changsha city. RESULTS: The general intellectual level of infants in Changsha city was good; stepwise regression analysis revealed the main possible risk factors affecting intellectual development of infants in Changsha city were newborn asphyxiated at birth (B = -17.162), perinatal fever in the first trimester (B = -8.084), infant with abnormal temper (B = -6.295) and poor appetite (B = -3.103). Parental care (B = 4.622) possibly benefit the infant intellectual development. CONCLUSION: These data suggested that the main measures to promote infant intellectual development were more attention need to be paid to health care for women and children, prevention of perinatal disease and infant rearing in a scientific way. PMID- 11860759 TI - [The trend of changes in the rate of admission and mortality among rural patients with acute myocardial infarction]. AB - OBJECTIVE: To evaluate the trend of changes in the rates of incidence and mortality of acute myocardial infarction (AMI) in rural population. METHODS: Analysis was made on the clinical data of hospital admission and mortality rate of patients with AMI in rural areas during 1976 and 1995 under every 5 - year interval: 1976 - 1980, 1981 - 1985, 1986 - 1990 and 1991 - 1995. RESULTS: The number of admission of patients with AMI in rural areas in these four periods were 82, 118, 177 and 299 cases respectively. The number of deaths were 17, 23, 30 and 39 cases and the fatality rates among inpatients were 20.7%, 19.5%, 16.9% and 13.0% respectively during these periods. The number of admission in females increased faster than that in males, the increase in percentage was 375.0% versus 207.4% (P < 0.05). In - patient fatality rate in females was significantly higher than that of males (19.8% vs 13.6%, P < 0.05). CONCLUSION: The rates of incidence and mortality of AMI have increased while inpatient fatality has decreased in rural population in the last 20 years. PMID- 11860760 TI - [Analysis on the epidemiology of syphilis from 1994 to 1998 in Beijing]. AB - OBJECTIVE: To explore associated factors related to the considerable increase of syphilis based on the epidemiologic analysis of new cases in the recent five years in Beijing, and to provide reference for further prevention and treatment of syphilis. METHODS: New cases of syphilis reported in the city were collected consecutively. RESULTS: It was indicated that the annual specific incidence rate had been greatly increasing, which was 3.81 per 100 thousand residents with an average annual increase as high as 387.68%. Most of the patients were cases in early phases. More second - stage cases were reported than first - stage cases. Among cases at first - stage, men were dominant seen while women were dominant among cases of second - stage. Patients aged 20 - 49 years counted for large percentage of the newly identified cases and the self - employed and the unemployed persons were seen in terms of job category. Married persons had a yearly increasing proportion in incident cases. Most popular way of transmission was through unmarried sexual contact. CONCLUSION: On the basis of the epidemiology feature of syphilis in Beijing, with prevention measures taken, the incidence of syphilis was controlled to some extent. Since syphilis is prone to the transmission of AIDS, the control of syphilis transmission has an important implication to the control of AIDS epidemic. PMID- 11860761 TI - [An epidemiological study on acute diarrhea during a filed exercise in PLA unit in an area in southern China]. AB - OBJECTIVE: To explore the epidemiological characteristics and risk factors of acute diarrheal disease during a filed exercise. METHODS: Epidemiological study of a certain artillery acute diarrheal disease during a filed exercise and multiple linear stepwise regression analysis, multiple stepwise discriminate analysis were used. RESULTS: Result showed that the incidence rate of training troops was significantly higher than that of the local troops at the corresponding period (7.32% vs 0.98% P<0.05). The incidence rate among officers was significantly lower than that of the soldiers (0.99% vs 8.20% P<0.05). The incidence rate was significantly higher at the beginning of assemble for training but smooth and steady afterwards with several outbreaks. There was highly significant difference in the incidence rate of 39 companies. Regression and discrimination analysis suggested that the incidence rate was correlated with the density of flies and average boiled water volume provided everyday. The correct classifying rate of the discrimination was 91.9% in total. CONCLUSION: These results provided a basis for prevention of acute diarrheal disease during filed exercise of military action. PMID- 11860762 TI - [A molecular epidemiological investigation on Leptospira interrogans serovar hebdomadis and australia in China]. AB - OBJECTIVE: In order to confirm whether cattle serve the source of infection for patients with Leptospira interrogans serovar hebdomadis and australia. METHODS: 12 field strains of leptospira interrogans serovar hebdomadis and australia were isolated from blood samples of patients with leptospirosis and farm cattle urine. An analysis of chromosome DNA restriction endonuclease patterns (REP) and restriction fragments length polymorphism (RFLP) of 16S + 23S rRNA gene were processed by digestion of chromosome DNA using EcoR I. RESULTS: The same serovar field strains of Leptospira interrogans from blood samples of patients with leptospirosis and farm cattle urine resulted in unique restriction endonuclease patterns (REP) and ribosomal types (RT). The chromosome DNAs from field strains that belonged to different serovars of Leptospira interrogans caused different restriction endonuclease patterns and ribosomal types. CONCLUSION: In accordance with this results, we recognize that farm cattle serve the infectious source of leptospirosis serovar hebdomadis and australia thus may have realistic significance in the control of leptospirosis serovar epidemic of hebdomadis and australia in China. PMID- 11860763 TI - [Study on the epidemiological features of Kawasaki disease in Jiangsu]. AB - OBJECTIVE: To investigate the incidence of Kawasaki disease and epidemiological characteristics in Jiangsu province. METHODS: Questionnaire and diagnostic guidelines for Kawasaki disease with color - printed photographs of typical clinical symptoms were sent to all county hospitals with pediatric beds. All the in - patients with Kawasaki disease hospitalized from January, 1993 to December, 1997 were studied regarding age, sex, season of illness, cardiac sequelae, outcome etc. RESULTS: There were 509 cases (329 males and 180 females) diagnosed with 489 cases (96%) clinically typical. The incidence of the 5 - year period was averaged 2.53 per 100,000 children under 5. The number of male patients with Kawasaki disease was higher than that in female. The age peak of incidence was between 6 month and 2 - year olds. 13.6% of all cases developed cardiac sequelae which mostly occurred in males and in those younger than 2 years of age. There was one patient recurred among total patients reported (recurrence rate 0.2%) and 2 patients died (fatality rate 0.4%). CONCLUSION: The epidemiological characteristics of Kawasaki disease in Jiangsu province were similar to those in Japan in sex, season distribution and cardiac sequelae. The number of patients in male was higher than in female. Patients were more commonly seen in spring than in winter. However, the incidence rate in Jiangsu was significantly lower than that in Japan and the age peak was between 6 month and 2 years old. PMID- 11860764 TI - [Epidemiological and clinical characteristics of Kawasaki disease in Shanxi province]. AB - OBJECTIVE: The objective of this paper is to describe the epidemiological and clinical characteristics of Kawasaki disease (KD) in Shanxi province, China during a 5 year - period from January 1993 to December 1997. METHODS: A province wide epidemiological survey on Kawasaki disease (KD) was carried ont by the China - Japan Kawasaki disease study group. The questionnaire form and the diagnostic criteria of KD which was prepared by the Japan Kawasaki disease research committee and translated into Chinese were sent of the departments of pediatrics of all the hospitals with 100 or more beds in Shanxi province. All the KD patients who were diagnosed during the observation period from 1993 to 1997 were asked to report to this survey. The database of reported KD in this survey was analyzed at the Department of pediatrics of the Shanxi provincial people's hospital, Xian, China. All the patients that satisfied the diagnostic criteria were included. RESULTS: A total of 105 (70%) hospitals responded and 376 cases of KD were confirmed. More cases were reported in 1993 and 1994. Of the total patients reported, 69% were children under 3 years old with male to female ratio of 1.6:1. The proportion of patients with cardiac sequelae was 19% with male to female ratio: 3:1. There were 4 fatal cases with a fatality rate of 1%. CONCLUSIONS: Since KD is common in China continuous surveillance is necessary to maintain high awareness of KD so as to identify possible risk factors and its association with other disease. PMID- 11860765 TI - [Analysis on Helicobacter pylori infection and its related diseases in 1,442 children in Tianjin city]. AB - OBJECTIVE: The aim of this study was to investigate the prevalence of Helicobacter pylori (Hp) infection in children in Tianjin and to study the relationship between the Hp infection and its related diseases. METHODS: (1) Serologic examination for Hp - IgG; (2) Rapid urease test for Hp (Hp - Rut); (3) Giemsa staining for Hp; (4) Gastric fluid examination for Hp by PCR. Hp infection were diagnosed by at least two criteria. RESULTS: (1) Of the 1 442 children, 651 were Hp - IgG positive. The total positive rate was 45.15%, Positive rates of Hp IgG among children under the age of 1 year, 3 years, 6 years, 10 years and to 14 years were 11.19%, 19.23%, 44.21%, 54.20% and 61.46%, respectively. (2) Positive rates of Hp - IgG were 31.18% (164/526) and 53.17% (48/916) in asymptomatic children and in patients with gastrointestinal symptoms. The rate was significantly higher in the later group (P < 0.005). (3) Hp infection rate in patients with duodenal ulcer (DU) was 90.76%, higher than that in other gastrointestinal diseases. (4) Histological examination showed that Hp infection rates were 33.20% and 83.97% in mild and moderate or severe gastritis, respectively. CONCLUSIONS: (1) The prevalence of Hp infection in children in Tianjin was high. (2) Rate of Hp infection increased with age and Hp infection rate was higher in children with gastrointestinal symptoms than with asymptomatie. (3) There was a high correlation in children with gastroduodenal diseases and Hp infection especially with DU. (4) Hp positive rate correlated with the pathohistologic severity of gastritis. PMID- 11860766 TI - [Study on the hepatitis virus infection among medical professionals]. AB - OBJECTIVE: In order to find out the present situation of hepatitis virus infection among medical professionals. METHODS: 140 medical professionals were studied on their serum HBsAg, anti - HBs, anti - HCV using EIA, and HEV - IgG, HGV - IgG with ELISA. RESULTS: The infection rates of HBsAg, HCV, HEV and HGV of medical professionals were 7.8%, 1.4%, 7.1% and 7.1% respectively with an overall infection rate of 23.5%. The infection rates for the clinical group and the non - clinical group were 33.3% and 12.3% respectively [with significant differences (P < 0.01)]. The infection rates of HEV and HGV for the clinical group were 8.0% and 12.0% respectively. It was found no superimposing infection in non - clinical group but was in clinical group. CONCLUSION: The result suggested that the infection rate of hepatitis virus in the medical professionals in the clinical group was higher than in the non - clinical group. PMID- 11860767 TI - [Study on the epidemiology of diabetes mellitus in peasants with different income in Binzhou prefecture Shandong province]. AB - OBJECTIVE: In order to study the morbidity of diabetes mellitus (DM), impaired glucose tolerance (IGT), and main effective factors of peasants with different income in poor area and to provide scientific data for strategy development. METHODS: The regions of investigation were divided into five parts: east, south, west, north and centre with two and three villages randomly selected for study from each region. Glucose - oxidase method was used to measure blood sugar and urine sugar. The classification and diagnosis standard were referred from the Beijing conference in 1980. RESULTS: The morbidity rates of non - insulin - dependent diabetes mellitus (NIDDM) and IGT were 1.21% and 2.35% in peasants who earned 800 and 2,500 yuan every year respectively. The standardized morbidity of NIDDM and IGT were 0.96% and 1.98%. We found that morbidity of NIDDM and IGT was very low among those younger than 20 years old but increased with age until reaching the top at the age of 70 to 80 years. DM patients were prominently seen at 30 to 40 year olds and IGT at 20 to 30 year olds. There was no difference of morbidity between male and female seen (chi(2) = 0.19 and 0.12; P > 0.05 and 0.05). The morbidity rates of DM and IGT in poverty type peasants who earned less than 1,200 yuan every year were 2.23% and 3.55% respectively, higher than those with adequate food and clothing type peasants who earned 2,000 to 2,500 yuan every year (morbidity rates were 0.39% and 1.18%; chi(2) = 18.11 and 18.10, P < 0.05 and 0.005) and those with common type of peasants who earned 1 200 to 1,900 yuan every year (morbidity rates were 0.80% and 1.92%; chi(2) = 25.85 and 18.20, P < 0.005 and 0.005). CONCLUSION: These results showed that sex did not obviously relate to morbidity of DM and IGT while age was one of the risk factors. The differences of income and unhealthy food intake played an role in the difference of morbidity to DM and IGT. It is important to increase the income of peasants and to change the unhealthy life style. PMID- 11860768 TI - [Epidemiological studies on the relationship between body weight and the prevalence rate of impaired glucose tolerance (IGT) in the population of urban areas of Tianjin city]. AB - OBJECTIVE: In order to explore the relationship between body weight and the prevalence rate of IGT in a community - based population and to provide scientific informations of IGT for prevention and control. METHODS: Cluster sampling was carried out in prevalence survey, together with clinical examination and test of blood plasma glucose. RESULTS: A sample of 62,577 people was performed by cluster random sampling. Results showed that the crude prevalence rate of IGT in the whole population of three communities was 13.13 per thousand in males, 19.18 per thousand in females, and 16.23 per thousand in both sexes respectively. The IGT prevalence rate of different body weight group in community population had a marked difference. There was a trand that the IGT prevalence rate increased markedly with the increase of body weight. CONCLUSION: The incidence of IGT is related to body weight and obesity. PMID- 11860769 TI - [Epidemiological feature on diabetes mellitus among permanent inhabitant in the Special Economic Zone of Shenzhen]. AB - OBJECTIVE: To investigate prevalence and relevant factors of diabetes mellitus among permanent inhabitant in Shenzhen. METHODS: The prevalence of 8 200 inhabitants aged over 20 in Shenzhen was studied. Blood glucose was determined by glucose - oxidase method. RESULTS: According to the WHO diagnostic standard, the prevalence rates of DM and IGT were 4.23% and 11.94% respectively. The prevalence increased apparently with age. The history of clan and overweight were related to the prevalence rates of DM and IGT. The people of DM and IGT were mostly accompanied with high levels of triglyceride, total cholesterol, uric acid, creatinine and low level of high dense lipoprotein (HDL). CONCLUSION: The history of clan, old age, overweight and higher fattiness were found risk factors of DM and IGT. PMID- 11860770 TI - [An observation of the immuno-persistence after inoculating with the domestic BRD II strain rubella vaccine among infants and young children]. AB - OBJECTIVE: To study the immuno-persistence after inoculating with the domestic BRD II strain rubella vaccine among infants and young children. METHODS: Hemagglutination inhibition (HI) antibody detection method was used to test on children at age 6 to 18 months without rubella infection or rubella vaccine immunization in Yantai city of Shandong province and were selected for the observation of the immuno-persistence against the domestic BRD II strain rubella vaccine from 1995 to 1998. RESULTS: Positive rates of HI antibody among children of 6, 7, 8, 9, 12 and 18 month-old of inoculation were 94.44%, 97.22%, 96.67%, 100.00%, 100.00%, 100.00% and 93.10%, 93.55%, 96.77%, 96.97%, 100.00%, 100.00% (P > 0.05), in one or two years respectively. The geometric mean reverse titers (GMRTs) were about 50% lower than that after 1 month after 1 year. Similar results were found that after 2 years GMRTs was about 50% lower than that after 1 year of inoculation. There was highly significant difference on GMRTs of HI antibody between infants with 6 to 7 month-olds (29.89) and young children with 8 to 18 month-olds (53.00) after 2 years of inoculation (t = 3.58, P < 0.001). CONCLUSION: The immunization schedule at the first dose for BRD II strain rubella vaccine should be started when the child is 8 month old. For the second dose, the immunization schedule used in other developed countries should be referred adopted in China. PMID- 11860771 TI - [Cost-effectiveness analysis on two different measles immunization strategies]. AB - OBJECTIVE: To seek the most economic and effective strategy in measles elimination in China, data of cost - effectiveness was analyzed to compare two different strategies of measles immunization (mass campaign and 2 - dose schedule) based on the data of population, economy and epidemiological features of measles in Henan province. METHODS: DALY was used as the measurement of effectiveness. Taking period from 1997 to 2020, the cost - effectiveness ratio (CER) was defined as the cost difference between mass campaign and 2 - dose schedule divided by the decline of DALYs if 2 - dose schedule had been substituted by mass campaign. RESULTS: Along with the increment of time, CER showed a descending trend then the mass campaign will have positive benefits after year 2013. CONCLUSION: Mass campaign is more cost - effective than 2 - dose schedule. PMID- 11860772 TI - [Primary study on immunologic effect of live attenuated hepatitis A vaccine (H2 strain) after booster dose]. AB - OBJECTIVE: To study the immunologic effect of live attenuated hepatitis A vaccine (H(2) Strain, 10(7.0)TCID(50)) after booster and to compare with the results of 1 dose live attenuated hepatitis A vaccine (H(2) Strain, 10(7.0)TCID(50)). METHODS: 42 susceptibles with negative anti - HAV were selected in Zhengding, Hebei province. Each subject received 3 doses live attenuated hepatitis A vaccine at 0, 2, 6 months and was bled at 1, 2, 6, 7, 9, 12 months after vaccination. RESULTS: The seroconversion rate at 1 month after the first dose was 81.4% and reached 100% after the second dose. GMT arrived the peak 2,739 mIU/ml at one month after the third dose, before stared declining. The seroconversion rate kept 100% at 12 months after the first dose, but GMT decreased to 979 mIU/ml. CONCLUSION: The first dose worked as the base of boostering. A booster dose of live attenuated hepatitis A vaccine could induce secondary immune response well. The immunologic effect after booster dose could match the effect with inactivated vaccine and was better than the results of 1 dose live attenuated hepatitis A vaccine. The program seemed to be useful to the protective effect and the immuno - persistence. PMID- 11860773 TI - [Immunohistochemical findings on liver tissue in patients with hepatitis C]. AB - OBJECTIVE: To investigate the distributions and infer relations of HCV antigens in the liver tissues. METHODS: Immunohistochemical techniques as polyclonal antibody to HCV, monoclonal antibody to HCV - NS3 and monoclonal antibody to HCV NS5 were used to detect HCV antigens in 107 patients with hepatitis C. RESULTS: HCV antigens in liver tissues were identified on these three kinds of antibodies. The positive rates were 38.3%, 28.0% and 52.3% respectively. The positive granules were located in the cytoplasma of liver cells. The distributions of positive cells were scattered, diffusive and clustered. No obvious anatomic association was observed between the expression of HCV - NS3 and inflammatory reaction. HCV - NS5 positive cells were more commonly seen around or in the flammatory focci. CONCLUSION: The expressions of various components of HCV were demonstrated in different concentrations. The expressions of HCV - NS5 might be related to the pathogenesis of hepatitis C. PMID- 11860774 TI - [Detection of Escherichia coli strains harboring pathogenicity island of Yersinia enterolitica in diarrheal patients and animals in China]. AB - OBJECTIVE: To investigate the prevalence of Escherichia coil strains harboring high pathogenicity island of Yersinia enterocolitica in diarrhea patients, animals, food samples and resulted clinical symptoms. METHODS: Colony hybridization, DNA dot hybridization and polymerase chain reaction (PCR) were used. RESULTS: HPI - harboring E. coli were found in the strains isolated in various provinces of China. The isolation rates were 27.05% (436/1,612) in the strains isolated from diarrhea patients, 10.23% (9/88) in the strains isolated from food samples and 5.71% (16/280) in the strains isolated from feces of cattle and pigs. Typical clinical symptoms of diarrhea caused by HPI - harboring E, coli were inappetency, abdominal pain, shiver and vapidity. Patient's temperature was usually normal or low. Over six episodes of diarrhea was frequently observed, of which most were unformed stools with mucous. CONCLUSION: E. coli strains harboring high - pathogenicity - island of Yersinia species were frequently isolated from diarrhea patients, animals and food samples as well, which might be one of the major causes of diarrhea in China. PMID- 11860775 TI - [Study on smoking pattern and related factors among residents aged over 15 years in Guangdong province]. AB - OBJECTIVE: To investigate the prevalence of smoking and related factors among residents aged over 15 years in Guangdong province. METHODS: Sampling survey, descriptive analysis and nonconditional logistic regression. RESULTS: Rates on general smoking, current smoking, regular smoking and heavy smoking among male residents aged over 15 years were 70.03%, 66.57%, 65.16% and 35.47% respectively and those among females were 3.04%, 2.67%, 2.54% and 1.12% respectively. There was significant difference between sex distribution. 86.71% of the male smokers started smoking at 15 to 29 years old, while the age distribution of noset of smoking among females was more scattered. Main reasons of starting smoking were to experience with social pressure and getting rid of feelings of fatigue. Tobacco products they often smoked included filter tip cigarette, handmade cigarette or dry cigarette. 52.42% of the smokers claimed that they often smoked in public places. 66.7% of the residents believed that smoking was either no harm or just minor while, 35.5% of them believed that smoking could get rid of fatigue and enhance the efficiency of work. Sex, age, occupation, marriage status and knowledge about the harm of smoking to health were factors related to smoking. CONCLUSION: Smoking has laid great harm to the health of residents in Guangdong province. Health education, stopping smoking in public places and no advertisement of cigarette under the management of laws are the best way to control the harm of smoking. PMID- 11860776 TI - [Establishment and initial utilization of an arithmetic model of chronic hepatic diseases]. AB - OBJECTIVE: An arithmetic model was established to reflect the natural development of chronic hepatitis B, and to help decision making in public health. METHODS: The frame of this model was composed of 6 states - chronic hepatitis B (CHB), natural recovery (NR), compensated cirrhosis (CIR), decompensated cirrhosis (DEC), hepatocellular carcinoma (HCC) and death (DEA). It was supposed that each state would develop into the other possible states at different rates after different years. According to many related reports, 15 values or functions of transfer rates were calculated, and on the ground of them the present model - the Foxpro program was established. Hypothetical CHB and other patients had been introduced to simulate the natural course over 55 years. RESULTS: It was shown that the observation period when 50% of NR, CIR, DEC and HCC were developed from CHB were 3, 11, 20 and 24 years respectively. The CHB, CIR, DEC and HCC patients could last 30, 12, 4 and less than 1 year respectively before half of them died. At the 30(th) year, proportions of death were 41.3% from HCC, 33.0% from DEC and 25.7% from the other causes. CONCLUSION: After the comparison between the main results of the model and real reports from the past years, it could be concluded that the model was reliable. PMID- 11860777 TI - [The serologic study on the blood transmission origin of HIV]. AB - OBJECTIVE: To study the origin of HIV blood spread in China. METHODS: Markers of HBV and HCV in 62 HIV - infections sera (27 blood donors and 35 i.v. drug users) were detected by EIA. RESULTS: The results show that the total HBV infection was 53.2% (33/62) and anti - HCV positive 95.2% (59/62). Super - infection of HIV, HBV, HCV was 51.6% (32/60) and super - infection of HIV, HCV and HIV, HBV were 27.4% (17/62) and 1.6% (1/62). Comparing the markers of HBV and anti - HCV between blood donors and i.v. drug users, there was no statistical significant difference (P > 0.05). The super - infection of HIV, HBV, HCV and HIV, HCV and HIV, HBV were also no statistical differences (P > 0.05) between two groups. CONCLUSION: Results indicated that a common infection mechanism might exist between Chinese HIV - infection blood donors and i.v. drug users. PMID- 11860778 TI - [Meta-analysis on the effectiveness of community-based multiple risk factor interventions for hypertension prevention]. AB - OBJECTIVE: To evaluate the effectiveness of multiple risk factor interventions for hypertension prevention in the community. METHOD: Twelve trials of multiple risk factor interventions for community-wide hypertension prevention according to the selective criteria were assessed by Meta-analysis, from 1970 to 1997. According to the heterogeneity of the studies, two different models, namely, Fixed Effect Model and Random Effect Model, were applied to analyze the results. RESULTS: The intervention effects of community-based hypertension control trials were estimated to be: a 1.75 mmHg (s(x) 0.57 mmHg) net decrease in mean systolic blood pressure, a 1.45 mmHg (s(x) 0.43 mmHg) net decrease in mean diastolic blood pressure in general population, 0.24 kg/m(2) (s(x) 0.08 kg/m(2)) and 0.32 kg/m(2) (s(x) 0.15 kg/m(2)) net decreases in body mass index among the total population and female population respectively, a 2.97% reduction in hypertension prevalence, and a 133/100,000 reduction in stroke prevalence in the Chinese population. The intervention effects for the treatment and controlled of hypertensive were significant to increase 25.07% and 40.45% in the rates and controlled of treatment respectively. No significant changes were noticed in the rates of smoking and hypertensive awareness in the general population. CONCLUSION: The results showed that the community-based multiple risk factor interventions for hypertension had positive impaction the reduction of risk factors to hypertension, thus to increase the effects on treatment and control of hypertension, decrease prevalences of hypertension and stroke. PMID- 11860779 TI - [Community -based comprehensive prevention and control of hypertension in China (CCPACH Study)-prevalence and epidemiological characteristics in urban and rural area]. AB - OBJECTIVE: To understand the epidemiological features and prevalence of essential hypertension in the rural communities. METHODS: A community-based survey on essential hypertension was carried out in urban and rural areas involving 280,000 population. RESULTS: The prevalence rate of hypertension in age 35 years and over was 31.7% and 32.9% in urban and rural respectively; the average age of the patients' in urbans was significantly higher than in rural areas. Among non hypertensive cases, more than 50% of the population having risk factors of hypertension in both urban and rural areas. Rates of treatment and control awareness of hypertension were 56.3%, 26.8% and 4.4% respectively in urban; comparing to 40.3%, 17.5% and 2.6% respectively, in the rural areas. More than half of the diagnosed patients before survey did not take antihypertensive medicine for treatment in both areas. The proportions the classification of hypertension were similar in both areas with majority of them, mild and isolated systolic hypertension. CONCLUSIONS: It was demonstrated that there is an urgent need to carry out comprehensive prevention and control for hypertension in urban and rural area. PMID- 11860780 TI - [The epidemiological characteristics of multiple metabolic disorders in hypertensive patients in the communities]. AB - OBJECTIVES: To describe the epidemiological characteristics of multiple metabolic disorders on hypertensive patients living in the communities. METHODS: The characteristics of metabolic disorders in the hypertensives from one community in Shanghai urban area were analyzed. Metabolic disorders would include dyslipidemia (high cholesterol and/or high triglycerides and/or low HDL cholesterol), diabetes and obesity. RESULTS: 59.2% of the hypertensives at least had one metabolic disorder. Proportionally, 83.9%, 15.5%, 0.6% of the patients would combine one, two, or all of the disorders. The percentages of patients combined with dyslipidemia, diabetes and obesity were 53.0%, 9.7%, 6.3%, respectively. 74.0% of patients with metabolic disorders were combined with dyslipidemia while 9.1% were combined with both dyslipidemia and diabetes. The standardized prevalence rate of male patients with metabolic disorders was 58.1%, higher than the rate of women (54.5%). The prevalence rate of postmenopausal female patients with metabolic disorders was 67.6%, significantly higher than that of the premenopausal women (44.4%) (chi(2) = 37.49, P < 0.01). CONCLUSION: Nearly 60% of the total patients combined with metabolic disorders. Dyslipidemia occurred in more than half of the patients in total. The proportion of senile patients with metabolic disorders was higher than the younger patients. Results suggested that attempts to maintain the coronary heart disease at low levels in the population should include treatment of dyslipidemia to keep lipid and diabetes prevalence at low levels other than regular antihypertensive therapy. PMID- 11860781 TI - [Evaluation on the effects of community-based comprehensive prevention and control of hypertension in the rural areas in China]. AB - OBJECTIVE: To explore the strategies and measures of community-based comprehensive prevention and control of hypertension in the rural areas. METHODS: Community-based comprehensive prevention and control of hypertension was put in practice in 120,000 people in Fangshan District of Beijing during the period of "The Ninth Five-Year Plan (1996 - 2000)". RESULTS: After intervention, blood pressure of the whole subjects were under control and kept the level of baseline data; the rate of increase of prevalence in the intervention areas was below the rate of that from 1979 to 1991 in China (the former was 14.7%, while the latter was 53.7%); the incidence rate of the subjects was 1.4%, close to the average rate of age 15 and above in China; the rates of awareness, treatment and control were above the levels of baseline data; SBP and DBP of the patients decreased by 2.9% and 5.6%, respectively; while the rate of having high risk factors reduced by 13.4% on average. CONCLUSION: The community-based comprehensive strategies and measures played an important and effective role in prevention and control of hypertension in rural areas. PMID- 11860782 TI - [Evaluation on the effect of Benazepril for hypertension through postmarketing surveillance]. AB - OBJECTIVE: To evaluate the efficacy and safety of Benazepril used among the essential hypertensives. METHODS: 1 831 essential hypertensive patients aged 35 to 75 were randomly selected from a community and followed for 18 months. The level of blood pressure, status of taking Benazepril and side effects were sequencially collected. RESULTS: 1/3 of the patients had taken antihypertensive drug before the study and the rate of compliance was over 96%. The effective rate of Benazepril was 73.6% at three months and increased to 84.7% at 18 months. Comparing with the baseline data, SBP and DBP declined 10.8 mmHg and 6.7 mmHg respectively. The rate of side effect was 22.7%. Cough was most commonly seen among side effects. The peak of first recording on side effect occurred at three months including 60% of them mild. CONCLUSION: Results showed that Benazepril had good efficacy and safety for the essential hypertension patients in a long-term observation. PMID- 11860783 TI - [A case-control study on the relationship between abnormal homocysteine metabolism and essential hypertension]. AB - OBJECTIVE: To examine the relationship between abnormal homocysteine metabolism and essential hypertension in a Chinese population, a community-based case control study was conducted. METHODS: 127 essential hypertensive patients aged 35 to 75 were randomly selected from a community. Another 170 control subjects with blood pressure < 140/90 mmHg were selected from the same community. Serum homocysteine was determined using HPLC. Folate and vitamin B(12) were measured by radioimmunoassay. MTHFR genotypes were identified by PCR and restriction fragment length polymorphism analysis with Hinf I digestion. RESULTS: After adjusting for age and sex, the mean homocysteine level was 10.56 micromol/L for hypertensive patients and 10.34 micromol/L for controls (F = 0.234, P = 0.63). No association between either SBP or DBP and Hcy concentration was found in subjects without anti-hypertensive medications. The prevalence of homozygousity for thermolabile MTHFR variant for this population was 13.1%, and the mutant allele frequency was 38.7%. There was no significant difference on genotype distributions and the mutant allele frequency for the two studied groups. However, the concentrations of folate and B(12) for the hypertensive subjects were generally higher than the controls. CONCLUSIONS: The present study did not discover that the elevation of Hcy levels or MTHFR mutation were independent risk factors for essential hypertension. The higher folate and B(12) in the hypertensive subjects might contribute to a lower risk. PMID- 11860784 TI - [An genetic epidemiology study of MTHFR gene and blood level (sibpair linkage analysis)]. AB - OBJECTIVE: To explore the relationship between MTHFR gene and blood pressure. METHODS: A total of 142 sibpairs were collected from rural communities of CCPACH. MTHFR gene genotypes were determined by an assay based on the polymerase chain reaction. Phenotype of this study was the level of blood pressure. Covariates were measured by questionnaire, physical examination and clinical indices. Sibpair linkage analysis was used to analyse data by SAGE software. RESULTS: In this study, prevalence of TT genotype of MTHFR gene was 9.3%, while CT was 41%. There was no evidence for linkage of the MTHFR gene with either systolic or diastolic blood pressure identified. CONCLUSION: In our study population, MTHFR gene did not seem to be not related to blood pressure. PMID- 11860785 TI - [Isolation and identification of HN-98 strain of spotted fever group rickettsiae]. AB - OBJECTIVE: One strain of rickettsiae was isolated from a patient with unknown fever from Qiongzhong county in Hainan Province in 1998, using embryonated hen eggs and proved to be the member of rickettsiae by the methods of morphology. It was named HS-98 strain after the name of the place and the year it was isolated. METHODS: The isolate was identified by methods of Microcomplement fixation assay and OCR/RFLP and compared with known species and strains of SFGR by DNA polymerase chain reaction and DNA polymerase chainreaction and was followed by restriction endonuclease fragment length polymorphism analysis. RESULTS: Results demonstrated that HN-98 strain was antigenically and genotypically identical to R. sibirica 246. CONCLUSION: The results suggested that there existed natural foci of spotted fever in Qiongzhong Area of Hainan Province. PMID- 11860786 TI - [A cohort study on the relationship between vegetable intake and risks of lung cancer in the Tin Corporation (YTC) miners in Yunnan]. AB - OBJECTIVE: To understand the relationship between vegetable intake and risk of lung cancer in the Tin Corporation (YTC) miners in Yunnan. METHODS: The reproducibility and validity of frequency of vegetable intake in Food Frequency Questionnaire (FFQ) were studied and the correlations between different vegetable intake and frequency and lung cancer relative risk among high risk population of lung cancer by Chi-square, single- and multi-Cox regression were analyzed. RESULTS: Results showed that the reproducibility and validity of wax gourd, cucumber, spinach, wild celery, fresh bush pepper, fresh soybean, sweet potatoes, asparagus, fresh bamboo shoot were good (r = 0.20 - 0.33, P < 0.05); There were significantly different incidences of lung cancer in different frequencies of wax gourd, cucumber, wild celery, fresh bush pepper out of 11 vegetables intake, and also in different total vegetable intakes (chi(2) = 8.83 - 30.64, P < 0.05). 11 vegetables and total vegetable intake were significantly negatively correlated with risks of lung cancer (RR < 1, P < 0.05). Only 2 vegetable frequencies were negatively correlated with risk of lung cancer by adjusting confounders such as age, occupational history, and smoking (P < 0.05). CONCLUSION: This suggested that vegetables intake may have a role in decreasing the incidence of lung cancer. PMID- 11860787 TI - [The relationship between angiotensin converting enzyme gene polymorphism and risk factors for coronary heart disease]. AB - OBJECTIVE: To explore the relationship between angiotensin converting enzyme (ACE) gene polymorphism and risk factors for coronary heart disease (CHD). METHODS: 148 patients with CHD and 159 unrelated healthy individuals were studied using polymerase chain reaction (PCR) and genetic methods. Risk factors of CHD were assessed by standard questionnaire, physical examination and blood tests. RESULTS: (1) There were no significant differences among genotypes in terms of years, sex, body mass index (BMI), systolic blood pressure, diastolic blood pressure, total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL), high-density lipoprotein-cholesterol (HDL), HDL(2), HDL(3), apolipoprotein (Apo) AI, ApoB, and lipoprotein (a) [Lp (a)]. The prevalence rates of hyperlipidemia, hypertension (HT), cigarette smoking, diabetes mellitus (DM) and obesity were similar in the ACE genotypes (P > 0.05). (2) The DD genotype frequencies in low CHD risk groups, as defined by ApoB < 125 mg/dl and BMI or = 500 mg/day, n = 106). The two groups were matched by age 8 month olds among the two types of residents. Using data originated from the hospital documents of 208 measles cases in 1998. RESULTS: Among the total 208 cases: (1) Of 107 cases of temporary residents in age > 8 mon., 79.4% were unvaccinated while 41.7% were unvaccinated among 72 cases of permanent inhabitants in age > 8 mon.; (2) The two peaks fell on 9 month to 5 year olds (32.7%), in which predominant cases were permanent inhabitant, while among the 14 - 36 year group (33.7%) temporary resident accounted for 71.4%; (3) Among 208 cases, there were 29 in 3 - 7 mon. olds, taking up 14.0% of the whole, who all did not receive the vaccinations. Seventeen of the 29 cases (58.6%) of the said children had mothers with positive histories of measles while the infants manifested atypical mild measles. The rest 12 of the 29 cases (41.4%) whose mothers had an negative histories of contracting measles, 4 infants manifested typical measles and 8 atypical mild measles. CONCLUSIONS: (1) One of the main causes of this epidemic was the unsound history of vaccination in the population; (2) In Shenzhen, preventive measures should focus on infants and young children among permanent inhabitants while focusing on adolescents and youths on temporary residents; (3) It was suggested that in Shenzhen, infants born to mothers without measles history or infants at the age borderline for measles immunization but without vaccination should receive it; Women in reproductive ages without measles history or vaccination should be considered as vaccination receivers. PMID- 11860816 TI - [A serological investigation of Legionella infection in eight-species of poultry and domestic animals in Sichuan province]. AB - OBJECTIVE: To understand the prevalence rates of Legionella infection in poultries and domestic animals in Sichuan province. METHODS: Serological investigation of antibodies against Legionella pneumophila serogroups 1 - 14 and Legionella micdadei was carried out, using microagglutination test (MAT) in eight species of poultries and domestic animals. RESULTS: Infections with multiple serogroups were identified. However, each species had its own major serogroup with positive rates of different serogroups ranging from 0.83% to 59.18%. Positive rates of the poultry and domestic animals were accounted for 7.73% and 4.25% respectively. CONCLUSION: It was suggested that poultries and domestic animals were widely infected with Legionella. It is important to carry out epidemiological surveillance in these domestic animals, poultries and environment for a better control program of this disease. PMID- 11860817 TI - [A case control study on the levels of lipid, fasting glucose, fasting insulin and sex hormones of hypertension in among elderly Uighur people in Hetian, Xinjiang]. AB - OBJECTIVE: To detect the relationship between the levels of lipid, fasting glucose, fasting insulin, sex hormones and hypertension in the elderly Uighur people in Hetian, Xinjiang. METHODS: A case control study on 105 cases of hypertension was performed, including 58 males and 47 females, aged 60 - 106. RESULTS: Among the lipid analysis items, the decrease of high density lipoprotein cholesterol (HDL-C) was the important factor related to the occurrence of the complications of hypertension (OR = 5.6214). The increases of both fasting glucose and fasting insulin levels were also risk factors of hypertension in the elderly Uighur people (OR = 2.4474 and OR = 2.5318). CONCLUSION: The risk factors of hypertension in the elderly Uighur people included decrease of HDL-C, high level fasting glucose and fasting insulin. PMID- 11860818 TI - [Analysis of clinical characteristic and tendency on inpatients with colorectal cancer in Shanxi Tumor hospital from 1989 - 1999]. AB - OBJECTIVE: To understand the clinical characteristics and variable tendency on inpatients with colorectal cancer admitted to Shanxi Tumor Hospital from 1989 - 1999. METHODS: Statistical analysis on all cases from 1989 - 1999. RESULTS: Results showed that 3,050 cases with colorectal cancer were admitted during 11 years including 2,246 (73.64%) cases of surgical patients. The 1,168 cases males and 1,078 females were with ratio being 1.08/1.0 (M:F). There was no obvious change on the numbers of inpatient from 1991 to 1994, but the numbers was higher than that in 1989 and 1990. The numbers of inpatient rose from 1995 to 1999 year by year. In all the admitted inpatients during the 11 years, 1,348 cases were from countryside and 898 cases from city with the ratio being 1.5/1.0. When adjusted by the general population in 1992, the frequency of the admitted patients was higher in city (14.66/10(5)) than that in countryside (5.96/10(5)). Regarding the distribution of inpatients, it was found higher in Tiayuan city and Jinzhong area than that in else areas. The age of inpatients rose from 40 years and reached high peak in 51 - 70 years. Dukes stages showed that most of inpatients were middle or advanced stage and chief ill location was in rectal (82.36%). Most histological classification referred to adenocarcinoma (56.23%) and papilla adenocarcinoma (20.88%). CONCLUSION: Data suggested that it is very important to strenghen the knowledge of prevention and self-protection of colorectal cancer. Screening and early diagnosis and early treatment should be paid great attention to improve survival rate among high risk population. PMID- 11860820 TI - [A comprehensive evaluation on psychological quality among automobile drivers]. AB - OBJECTIVE: To study the comprehensive evaluation on the indexes of psychological quality on automobile drivers. METHODS: A index system, consisted of seven psychological indexes (including relevant physiological indexes), was developed through Delphi method and mathematical statistics. Basic data of the seven psychological and physiological indexes including mean, standard deviation and percentile, were taken from more than 8,000 occupational licensed automobile drivers and more than 6,000 trainees who were randomly sampled from 5 cities across China. The weight coefficients of indexes were determined by analytical hierarchy process (AHP). RESULTS: Comprehensive evaluation equations of the occupational automobile drivers and the trainees were established respectively. The critical values of evaluation were divided into grades by the percentiles of evaluated values. Verified results showed that there was significant difference of the values under evaluation between drivers who had accidents and who did not (t = 14.68, P < 0.01; H = 73.61, P < 0.001). The test of consistence showed that the consistence was quite good (rate of consistence = 92.99%; kappa = 0.85). CONCLUSION: Results showed that the comprehensive equations for evaluation were reasonable and feasible for detecting the potential accident-proneness drivers. PMID- 11860819 TI - [A 1:2 matched case-control study on the risk factors of uterine myomas]. AB - OBJECTIVE: To investigate the risk factors and their association with Uterine myomas. METHODS: A 1:2 matched case-control study was carried out in Guangzhou including 211 cases and 422 controls. Data were analyzed with conditional logistic regression. RESULTS: Results showed that Pelvic inflammatory, Erosion of Cervix, Menopathy, Adnexitis and family history of Uterine myomas were identified as risk factors, with odds ratios of 3.101, 2.904, 3.942, 7.742 and 2.426 respectively while delayed menstruation was proved to be a preventive factor, with odds ratios of 0.841. CONCLUSION: Uterine myomas seemed to be related to Pelvic inflammatory, Erosion of Cervix, Adnexitis, Menopathy and family history of Uterine myomas. PMID- 11860821 TI - [The relationship between polymorphism of angiotensinogen gene and essential hypertension]. AB - OBJECTIVE: To study the renin-angiotensin system (RAS) involvement in the hormonal regulation of BP and blood volume and the relationship between angiotensinogen (AGT) was the sole preliminary acting materials in the RAS. METHOD: Population-based case-control study was conducted using PCR-RFLP techniques to explore the relationship between the M235T polymorphism of AGT and EH in Wangxin township, Jiading District of Shanghai. RESULTS: The frequency of TT genotype in EH cases was significantly higher than in controls (42.8% and 33.0%). The odds ratio (OR) for those exposed to TT genotype was 3.61, comparing with MM genotype. It was suggested that TT genotype be the susceptible genotype of EH in Wangxin. The frequency of allele T in EH cases was 73.89%, significantly higher than that of 60.31% in controls. With the increase of allele T, the risk of EH also increased (P < 0.05). In EH cases, the BMI increased in the order of MM, MT and TT, and significant differences between the groups were observed. CONCLUSION: TT genotype is the susceptible genotype of EH in Wangxin. PMID- 11860825 TI - [Relationship between CYP1A1, GSTM1 genetic polymorphisms and susceptibility to esophageal squamous cell carcinoma]. AB - OBJECTIVE: To investigate the association between susceptibility of esophageal squamous cell carcinoma and the genetic polymorphisms of CYP1A1 and GSTM1. METHODS: Subjects were comprised of 107 esophageal cancer patients and 111 healthy controls. Genotyping of both CYP1A1 and GSTM1 were performed in cancer tissues of all 107 patients and peripheral blood leukocytes taken from the controls by polymerase chain reaction. RESULTS: There were no significant differences in the frequency distribution of CYP1A1 polymorphisms between esophageal cancer patients and healthy controls although the frequency of CYP1A1 with at least one allele of Val showed slightly higher in individuals with esophageal cancer. However, significant difference was observed in the frequency of GSTM1-nulled individuals with esophageal cancer comparing with the controls (P < 0.05). When subjects were categorized by both CYP1A1 genotype and GSTM1 genotype, GSTM1 (-) became markedly expressed in patients with CYP1A1 (I/I) than in the corresponding controls (67% versus 40%, P < 0.01). The frequency of CYP1A1 genotype with at least one allele of Val (I/V and V/V) was also statistically higher in patients with GSTM1 (+), comparing to the corresponding controls (64% versus 41%, P < 0.05). CONCLUSIONS: It was suggested that: genetic polymorphisms of CYP1A1 and GSTM1 were susceptible to esophageal cancer; individuals who are GSTM1-null have an increased risk of developing esophageal cancer; individuals with combined CYP1A1 (I/I) and GSTM1 (-) or with combined CYP1A1 (I/V, V/V) and GSTM1 (+) were more susceptible, when comparing to those with combined CYP1A1 (I/I) and GSTM1 (+). PMID- 11860824 TI - [Study on the relations between HLA-DRB 1 alleles and Helicobacter pylori infection]. AB - OBJECTIVE: In order to study the relation between human leukocyte antigen (HLA) DRB1 alleles and Helicobacter pylori (Hp) infection. METHODS: Hp-IgG antibody from 46 gastric cancer (GC), 75 esophageal cancer and 100 population-based controls were identified by Hp-IgG quantitative enzyme immunoassay. Biotest HLA DRB enzyme linked probe hybridization assay kit (low resolution) was used to identify DRB1 alleles. RESULTS: (1) Frequency of DRB1 * 08 was significantly higher in Hp-IgG positive group than in Hp-IgG negative group (13.1% vs 4.4%, chi(2) = 11.14, P < 0.001). Frequency of DRB1 * 12 was significantly lower in Hp IgG positive group than in Hp-IgG negatives (5.4% vs 11.3%, chi(2) = 4.49, P < 0.05). (2) Frequency of DRB1 * 02 in GC was significantly higher than that of controls. Frequency of DRB1 * 07 in GC was significantly lower than that of controls. However, neither the frequency of DRB1 * 02 between Hp-IgG positive and Hp-IgG negative groups nor the frequency of DRB1 * 07 between Hp-IgG positive and Hp-IgG negative groups showed significant differences in GC and controls. CONCLUSIONS: (1) HLA-DRB1 * 08 might serve a genetic risk factor for Hp infection while DRB1 * 12 might play a role of protecting effect against Hp infection. (2) DRB1 * 02 might be a genetic risk factor for GC while DRB1 * 07 might play a role of protecting effect against GC. However, the relations between DRB1 * 02, DRB1 * 07 and GC were not associated with Hp infection. PMID- 11860823 TI - [Polymorphisms of chemokine receptor alleles influencing genetic susceptibility to HIV-1 infection in Mongolia population in China]. AB - OBJECTIVE: Mutant frequency and polymorphism of HIV-1 resistance CCR5-Delta32, CCR2b-64I and SDF1-3'A alleles were investigated in Chinese population from Mongolian ethnic origin. METHODS: Whole blood samples from 134 Mongolian subjects were collected randomly and their genomic DNA were extracted using Qiagen Blood Kit. Allelic frequency was identified by means of PCR or PCR-RFLP analysis. Allelic polymorphism in population and between sex in the sample as well as correlation of the three genes were analyzed by chi(2) test. RESULTS: The frequencies of the three alleles were as following: CCR5-Delta32 1.1%, CCR2b-64I 24.8% and SDF1-3'A 22.0% respectively. Distribution of the three mutant alleles among the Mongolian population was in accordance with Hardy-Weinberg equilibrium. Statistical analysis showed there was a higher frequency of CCR2b-64I in female than in male subjects (29.2% vs 19.7%). No Statistical difference was found in the allelic frequencies of both CCR5-Delta32 and SDF1-3'A between male and female individuals. CONCLUSION: Compared with the Caucasian American, there were higher frequencies of CCR2b-64I and SDF1-3'A alleles and lower frequency of CCR5-Delta32 allele found in Mongolian population while the factors responsible for the variation of genetic polymorphisms in different ethnic populations need to be clarified. PMID- 11860822 TI - [Using multiplex PCR for the detection of virulence genes in Escherichia coli O157:H7]. AB - OBJECTIVE: To detect and characterize the virulence genes in E. coli O157:H7 isolated from various reservoir in six areas of Jiangsu province. METHOD: The virulence genes of Shiga-like toxin (SLT(1) and SLT(2)), intimin (eaeA) and hemolysin (hlyA) were chosen as the target genes and amplified in multiplex PCR assays. RESULTS: Of the eighty-five E. coli O157:H7 strains, the overall virulence gene prevalence was found to be 56.5% (48/85). The prevalence rates virulence genes of isolates from various areas were different from 0% up to 90.5%. It seemed to exist a relationship between the virulence gene prevalence and the level of incidence. In the areas where rates of incidence were divided into high, low, sporadic or zero, the prevalence rates were 85.7% (36/42), 52.6% (10/19) and 8.3% (2/24), respectively. The prevalence rates of isolates were also different from various reservoirs, decreasing by sheep, cattle, pig and poultry. One isolate from a rabbit was positive for SLT(2), eaeA and hly genes. Of forty eight isolates carrying virulence genes, 38 (79.2%) had SLT(2), eaeA and hly genes, taking the dominate virulence gene pattern, 8 (16.6%) had all of the four virulence genes 2 (4.2%) had both SLT(2) and hly genes respectively. In addition, SLT(1) gene showed a lower prevalence, which was different from some findings abroad. CONCLUSION: Since virulence gene pattern of E. coli O157:H7 is an important molecular epidemiological marker, it can provide an useful information for epidemiologic studies, and helpful to the design of prevention and control strategies. For virulence gene detection, multiplex PCR seems to be a simple, rapid, specific and sensitive method. PMID- 11860826 TI - [Study on the application of random amplified polymorphic DNA in Yersinia pestis genotyping]. AB - OBJECTIVE: To study the type of 103 Y. pestis strains that isolated from different plague foci and different ecological types. METHOD: Random amplified polymorphic DNA (RAPD) was used. RESULTS: Y. pestis were divided into two RAPD types: most strains that isolated from elsewhere in the country belonged to RAPD 1 type, while most strains that isolated from Qinghai province were RAPD-2 type. CONCLUSION: The genome structures of different ecotype Y. pestis were different providing the foundation of further research and prevention of plague. PMID- 11860827 TI - [Homogeneity study on the Streptococcus suis isolated from human and swine]. AB - OBJECTIVE: To identify S. suis and to evaluate the homogeneity of isolates of S. suis from human and swine. METHODS: Culture, morphology, API biochemical tests and serum coagulate tests were used. All the 7 strains of streptococcus were confirmed to be S. suis serotype 2. Two strains and 1 strain were isolated from blood and cerebrospinal fluid (CSF) of patients, respectively, while the other 4 strains were obtained from the infected swine or their corpses. Seven strains of S. suis serotype 2 and reference strain SS2 were analyzed by randomly amplified polymorphic DNA analysis with six primers and Rep PCR with four primers and thalli fatty acid profile analysis. Cluster and principal compound analysis of results were performed with RAPD, Phylip and Treeview software. RESULTS: By analysis on RAPD patterns, a close relationship among 7 strains of S. suis serotype 2 and reference strain SS2 was discovered. Human-born strains and swine born strains exhibit similar RAPD patterns. There was a same clonal relationship between strains originated from the blood and CFS of the patients. These results were confirmed by thalli fatty acid profile to have genotypic and phenotypic identity. CONCLUSION: Homogeneity exists among the 7 strains and reference strain SS2. PMID- 11860828 TI - [Randomized controlled trial of sequence mass screening program for colorectal cancer]. AB - OBJECTIVE: In order to assess the effectiveness of mass screening program for colorectal cancer, a sequence mass screening program based on RPHA-FOBT and individual quantitative risk assessment model (attributive degree value, AD) was used and evaluated on its effectiveness in a randomized controlled trial. METHODS: The residents of Jiashan county aged 30 years and over were randomized to either screening or control groups in 1989. Participants in screening group were asked to fill in a questionnaire and to submit one paper slide with stool. Participants who tested positive underwent diagnostic evaluations including flexible sigmoidoscopy and colonoscopy. RESULTS: According to the cancer registry of Jiashan, after initial mass screening in 1989, the 8-year cumulative incidence per 1,000 of colorectal cancer in screening and control groups appeared to be 3.95 (95% CI 3.81 - 4.10) and 4.01 (95% CI 3.86 - 4.16) respectively. There was no significant statistical difference between two groups (P > 0.05). Nevertheless, the 8-year cumulative mortality for colorectal cancer in screening group (2.08 per 1,000; 95% CI 1.96 - 2.18) was reduced 14.7% comparing with the control group (2.44 per 1,000; 95% CI 2.33 - 2.55). In particular, the cumulative mortality of rectal cancer was significantly (31.7%) lower than that in control group. Log-rank test showed that survival rate of rectal cancer in screening group was higher than that in controls (log-rank = 9.01, P = 0.0027). CONCLUSIONS: The sequential mass-screening program which based on RPHA-FOBT and ADV might reduce the mortality for colorectal cancer in the Chinese population. PMID- 11860829 TI - [A case-control study on the risk factors of esophageal cancer in Linzhou]. AB - OBJECTIVE: To explore the characteristics of prevalence and influencing factors on the genesis of esophageal cancer. METHODS: A population-based 1:1 matched case control study was conducted in Linzhou. A total number of 352 pairs of cases and controls matched on sex, age and neighborhoods. Data was analysed by SAS software to calculate the odds ratio of and to evaluate the relative risks. RESULTS: It was found that lower socio-economic status, environmental pollution around the residential areas, lampblack in room, lower body mass index (BMI), more pickled food intake, cigarette smoking, alcoholic drinking, vigor mental-trauma and depression were risk factors of esophageal cancer. It also showed that the subjects having had history of upper digestive tract operation, dysplasia of esophagus and family history of carcinoma markedly increased the risks of developing esophageal cancer. CONCLUSION: Esophageal cancer seemed to be resulted from the combination of genetic and environmental factor, hence called for of medical surveillance and comprehensive prevention. PMID- 11860830 TI - [A cross-sectional study on the effects of microcystin in drinking water to the health of human beings]. AB - OBJECTIVE: To understand the effects of microcystin (MC) in drinking water on health of human beings. METHODS: In Taixing, a cross-sectional study was conducted in two middle schools with different types of drinking water, from which 111 and 92 students were randomly selected and investigated. Blood samples from all individuals were collected and tested for serum HBsAg, HBeAg, anti-HBe, anti-HBs, anti-HBc with enzyme linked immunoabsorbent assay and for serum alanine aminotransferase (ALT), gamma-glutamyltransferase (gamma-GT) and alkaline phosphatase (ALP) contents by colorimetry. These levels of enzymes in sera were compared after the students were stratified according to the status of HBV infection. RESULT: With the exclusion of the effect of HBV infection, three enzymes levels in sera between two groups of students showed significant differences (P < 0.01), much higher in exposure group than those in control group. CONCLUSION: MC was possibly harmful to human beings in drinking water and induced the contents of some enzymes from the liver of human beings to increase in sera. PMID- 11860831 TI - [Hostility behavior as risk factor of coronary heart disease]. AB - OBJECTIVE: To explore the relationships between hostility, and the risks of coronary-prone behavior and the clusters of unhealthy behaviors in individuals with different levels of hostility. METHODS: A cross-sectional study was carried out among 1 010 male employees from two factories in Beijing Capital Steel and Iron Company in 1997. Data on major risks of cardiovascular, other social demographic factors and levels of hostility by Cook-Medley Scales were collected. RESULTS: Body mass index, amounts of cigarette smoking and alcohol drinking were significantly increased (P < 0.01) with a characteristic of reduced social support from one's colleagues, when individual's level of hostility increased. In addition, less years of education, higher prevalence of other chronic diseases (P < 0.05) and clustering of various unhealthy behaviors were found related to the increase of individual's hostility. The odds ratio in the group with highest hostility was 2.37 times higher than in the lowest one, after confounding effects from age, blood pressure, education level, status of chronic disease and social support being adjusted. No significant differences on both systolic and diastolic blood pressures among groups with different levels of hostility were discovered. CONCLUSION: The association between hostility, coronary-prone risk behavior and cluster of unhealthy behaviors suggested that hostility might increase the risk of coronary heart diseases through affecting individual's behavior. PMID- 11860832 TI - [Safety and immunogenicity of inactivated bivalent EHF vaccine in humans]. AB - OBJECTIVE: Safety and immunogenicity of inactivated bivalent EHF vaccine in humans were evaluated in the epidemic area of Zhejiang province, China. METHODS: Susceptible persons with negative anti-EHF were selected in Jiande county, Zhejiang province to receive 3 doses of inactivated bivalent EHF vaccine at 0, 7, 28 days. A booster injection was given one year after the primary immunization. Antibody responses were measured in human volunteers by IFA and MCPENT. Local and general reactions were recorded within 72 hours after each vaccination by physicians. RESULTS: Two weeks after the primary vaccination, 99.04% of the subjects developed significant hantavirus antibody titre measured by IFA which had a 37.34% drop one year after the primary vaccination. Seroconversion rate increased to 100% two weeks after the booster dose. Neutralising antibody titres paralleled this trend with 100% of vaccine recipients producing neutralising antibody two weeks after the primary doses. However, it dropped to 80% one year after the primary vaccination. One hundred percent of the vaccine recipients started to respond two weeks after boosting. The geometric mean titre (GMT) of neutralising antibody against 76 - 118 and UR were 18.27 +/- 2.21 and 12.47 +/- 2.16 respectively after the primary injections, but it increased to 37.09 +/- 2.24 and 32.61 +/- 2.05 respectively after the secondary immunization. General and local reaction rates were 0.46% and 1.98%, with no severe side effects observed in the vaccinees. CONCLUSION: The vaccine was well tolerated and could induce good humoral immune response. PMID- 11860834 TI - [Analysis on the present situation of human rabies epidemic in China]. AB - OBJECTIVE: To summarize and analyze human rabies epidemic situation in the last five years in China and to explore the possible factors causing the rising of epidemics to provide reference for the prevention and control of rabies in the future. METHODS: Collecting and analyzing data of rabies epidemics in the last five years in whole nation. RESULTS: Human rabies is continuing going up in the last five years in China with an increase of 44.65%, 1.74% and 45.93% in 1997, 1998, 1999. Human rabies incidence was also seen an increase from 0.0134/100,000 in 1996 to 0.02708/100,000 in 1999. The provinces of rabies increase were mainly located in south China. Among them, Jiangsu province reported 40 human cases in 1999, which increased 207.69% than the corresponding period in 1998. The cases in these high epidemic provinces took up 75% of the whole nation's cases. Number of reported rabies case was also rising. CONCLUSION: Facts as increasing number of dog raising, having not been injected with rabies vaccine and antirabies serum timely or full course after exposure, unstrictly controlling the quality of rabies vaccine as well as lacking good cooperation in different official department on rabies control might serve as factors responsible for the recurrent increase of human rabies. PMID- 11860833 TI - [Report on epidemic hemorrhagic fever (EHF) surveillance in 1999]. AB - OBJECTIVE: To understand the epidemic tread of epidemic hemorrhagic fever (EHF) in 1999 and to develop programs for its control and prevention. METHODS: Epidemiological, zoo-epidemiologic, serologic and viral diagnostics were used to analyse the information of monthly and annual reports from surveillance spots. RESULTS: Twelve thousand nine hundred and thirty cases of EHF in all surveillance spots were reported with 135 cases of death in 1999. The morbidity and mortality were 5.37/100,000, and 1.04% respectively, which had a 1.47% decrease and 9.47% increase, comparing to that of 1998. The accordance of serological and clinical diagnosis was 67.25%, with rate of misreporting 1.28%. In animal surveillance, the density of mouse was 7.46% with predominant species Apodemus agrarius. In residential areas, the density of mouse was 6.21%, with predominant species Rattus norvegicus. For Apodemus agrarius in fields and Rattus norvegicus in residential areas, the indexes of HV carrier were 0.032 and 0.031 respectively in 1999. In spring and autumn, the indexes of HV carrier in Rattus norvegicus were apparently higher than that of Apodemus agrarius as the indexes of HV carrier in Rattus norvegicus were 0.028 and 0.036 comparing with 0.026 and 0.020 in Apodemus agrarius. CONCLUSION: It is suggested that more attention should be paid to the surveillance in low and medium-sized epidemic areas because northern China is dry with short rainy season while the epidemic situation in the serious epidemic areas had been under control to some degree. PMID- 11860836 TI - [Study on the epidemiological feature and control measure for rabies from 1990 to 2000 in Jiangsu province]. AB - OBJECTIVE: To analyze the epidemiological feature of rabies and explore the risk factors in Jiangsu for the control of rabies. METHODS: Data on rabies in Jiangsu from Jan. 1990 to Sep. 2000, and on case studies as well as surveillance from 1998 to 2000, were collected. Circular distribution statistics and Chi-square test were used. RESULTS: The number of rabies had been decreasing since 1990 yearly, but increased in the last two years. The new cases of rabies tended to focus in the area of Nantong, Yanchen and Taizhou city. There were about 80% of cases in the above areas. Dogs were the main hosts (93.22%). Ninety-four point nine two percent of the persons were affected through dog bites with major wounded part as thoracic limbs (46.27%). Rates of severe bites, being wounded close to the central nervous system and more than 1 wound per cases were significantly higher than that in the population under surveillance. The antibody conversion rate was 86.28% after immunization in the surveillance population. The immunization rate was 58.20% among dogs. CONCLUSION: Severe deep bites, wounded part close to the central nervous system and more than 1 wound per case were the risk factors in patients with rabies. Maltreatment and low immunization rate were important risk factors as well. PMID- 11860835 TI - [A study on the epidemiological characteristics and the preventive measures of rabies in Shanghai]. AB - OBJECTIVE: To analyze the epidemiological characteristics and the preventive measures of rabies in Shanghai from 1950 to 2000. METHODS: Suspected cases of rabies were investigated epidemiologically and collection of samples to isolate the virus. RESULTS: The most highest morbidity of rabies was 1.98/100,000 in 1950. There had been neither case of rabies, nor the rabies virus positive identified in dogs between 1958 and 1988. The morbidity of rabies was 0 - 0.03/100,000 from 1989 to 2000. According to the data of 20 reported cases from 1985 to Sept. 2000, the mean incubation was 66.37 days, except one case of 14 years. The mean course of diseases was 4.55 days. Seventy percent of the cases did not receive any preventive measure. A hundred and ninety-four persons who were bitten by rabies virus positive dogs were timely treated and all of them survived. In recent years, there has been more than 50,000 persons bitten by dogs every year in Shanghai. The rate of rabies vaccine immunization was over 95%. The rate of anti-rabies serum immunization was over 20%. CONCLUSION: Rabies is a preventable disease even when persons were bitten by dogs with rabies virus positive. The important measure is to treat the wound timely, giving vaccine and anti-rabies serum injection. According to the regulation of dog control in Shanghai, the number of dogs be kept at the lowest level, immunizing all the registration dogs, and destroying the none-registration dogs are the most essential measures to be taken. PMID- 11860837 TI - [Studies on the manufacture and immunogenicity of purified rabies vaccines on humans Vero cell]. AB - OBJECTIVE: Using Vero cell as basic cultural material to improve the quality of rabies vaccines and to produce rabies vaccines for humans. METHODS: CTN-1V10 strain were used for production. Vero cell of 150th generation were used for cultivation. Rotating cultivatal method with rotating bottle was used. Fluids with virus were collected at different time. Puritied rabies vaccine was produced on Vero cell after clarification, condensation, purification and extermination. A batch of vaccines made by this techniques were used for immunological observation. Sixty-three people were injected with this rabies vaccine according to the procedure of time of exposure. Thirty of them were injected with vaccines made in France (Verorab) while the others were injected with vaccines to be tested. Side effect and neutralizing antibody were recorded. RESULTS: The quality of this newly developed rabies vaccines has met the quality set by WHO. After all dosages of injection, the rates of positive antibody were both 100% in two groups. The neutralizing antibody among testing group was 11.94 IU/ml comparing with control as 11.69 IU/ml. CONCLUSION: Purified rabies vaccines on Vero cell for humans had reasonable manufacture technique and less little effect with good technological imnunogenicity. PMID- 11860839 TI - [The relationship of low density lipoprotein receptor gene polymorphism and hyperlipidemia]. AB - OBJECTIVE: To study the relationship of low density lipoprotein receptor gene polymorphism and hyperlipidemia in the population with essential hypertension. METHODS: People with different lipid levels including 107 hyperlipidemia, 104 at margin level and 108 normal were recruited in the study. Their polymorphisms of LDL-R gene were analyzed using PCR-RFLP. RESULTS: There were three kinds of genotype: (+/+), (+/-), (-/-). In male, the frequencies of the (+/-) in three study groups were shown as follows: 41.18% in hyperlipidemia, 46.15% in margin level, 19.05% in normal lipid. The frequency of (+) allele was significantly higher in hyperlipidemia than that in normal lipid (24.51%, 25.00% and 11.11%, respectively). In women, the differences were not statistically significant. The nonconditional univariate and multivariate logistic regression analysis demonstrated that (+) allele of Ava II polymorphism of LDL-R was a genetic marker of male's hypercholesterolemia. CONCLUSIONS: The frequency of (+/-) hyperlipidemia in males was higher than that in normal lipid group and the (+) allele in male hyperlipidemia was significantly more frequent seen than that in normal lipid group. These results suggested that polymorphisms of LDL-Rgene might play an independent role of risk factor for hyperlipidemia. PMID- 11860838 TI - [Study of triglyceride distribution aged 35 - 64, and their association with other cardiovascular disease risk factors in 11 provinces]. AB - OBJECTIVE: Study of triglyceride distribution and their association with other major cardiovascular disease risk factors in 11 provinces. METHODS: Twenty-seven thousand and sixteen persons in 11 provinces, aged 35 - 64, were screened for cardiovascular disease risk factors. RESULTS: (1) Results of plasma triglyceride level distribution showed that there was significant difference among 11 province, the highest/lowest ratio was 2.3. (2) Plasma triglyceride level was higher in men than in women (P < 0.001). (3) The prevelance of hypertriglyceridemia was the most frequent type of the dislipidemia. (4) Multiple stepwise regression analysis showed that total cholesterol, high-density lipoprotein cholesterol, body mass index, blood pressure, blood glucose and physical activity, significantly and independently correlated with hypertriglyceridemia prevelance. (5) There existed aggregation of risk factors in hypertriglyceridemia. Seventy-one point zero percent of hypertriglyceridemia had at least one other risk factors in men and 74.4% in women, while it was 53.8% men and 52.2% women of those with non hypertriglyceridemia (P < 0.01). CONCLUSION: Our date indicated that fasting triglycerides were associated with more arteriosclerosis factors and could serve as a marker for several cardiovascular disease risk factors. More patients with hypertriglyceridemia combined with other situation of metabolic syndromes thus might have higher risks. PMID- 11860840 TI - [Current status and distribution of deafness in the elderly in several cities in China]. AB - OBJECTIVE: To survey the current status and distribution of deafness and its effect on daily life activities in the old population. METHODS: Eight thousand two hundred and fifty-two elderly aged 60 years and above in the urban and rural areas of Beijing, Shanghai, Guangzhou, Chengdu, Xi'an and Shenyang were investigated, using a clustered random sampling methods. RESULTS: Overall crude prevalence of deafness in the elderly was found to be 33.7%, but the self reported crude prevalence was low, only 47.1% when shown by medical examination. Crude prevalence rates were increasing with ageing, with 21.6%, 30.0%, 35.6%, 42.6%, 55.5% and 61.6% respectively (P < 0.01) in the age groups of 60-, 65-, 70 , 75-, 80-, 85- years old. Rate of deafness was highest in Beijing (58.5%), among the in-house workers (48.9%) and the lowest in scientists, teacher and health workers (28.5%). Logistic regression analysis showed that the crude prevalence was related to ageing, profession and area (P respective < 0.01). CONCLUSION: The prevalence of deafness was high in the elderly which increased with ageing with different areas, professions and the level of education. It was lower when self reported than shown by medical examination. Prevention and treatment of deafness in the elderly should be strengthened. PMID- 11860841 TI - [The survey of HIV prevalence among children between 0 to 7 years old in one county in China]. AB - OBJECTIVE: To estimate the HIV prevalence among children between 0 to 7 years old in one county with HIV epidemic. METHODS: In September 2000, finger tip blood samples were collected from 619 children between 0 to 7 years old in three districts of one county with high HIV prevalence and tested for HIV antibody and HBV antigen. RESULTS: Among those 619 children, 11 were found HIV positive, with the prevalence of 1.8% (11/169). The HIV prevalence of three districts were 5.9% (10/169), 0.5% (1/206), and 0% (0/244), respectively. The possible channel associated with the HIV infection was mother-to-child transmission since none of the infected children had the history of blood transfusion. Twenty-four tested children were found HBV positive and the prevalence was 4.3 (24/619). CONCLUSION: There was higher HIV prevalence in children between 0 to 7 years old in certain areas. More intervenes are needed in the future to prevent the HIV spreading through mother-to-child transmission. PMID- 11860843 TI - [Logistic regression analysis of wedge-shaped defects in the middle-aged and elderly]. AB - OBJECTIVE: To investigate and identify the possible clinical risk factors associated with the formation of wedge-shaped defects. METHODS: Four hundred and ninety-three subjects were examined. The possible risk factors associated with wedge-shaped defects were identified by logistic regression analysis. RESULTS: The prevalence of wedge-shaped defects was observed 90.5% in this population. The prevalence and severity of wedge-shaped defects were found to be increasing with age. The correlation between age, sex, preferred side for tooth chewing, brushing twice a day, malocclusion, the formation and severity of wedge-shaped defects were noted. CONCLUSION: Occlusal factors and tooth brushing have found related to the formation of wedge-shaped defects. PMID- 11860842 TI - [Observation on the effect of disinfection to HBsAg by electrolyzed oxidizing water]. AB - OBJECTIVE: Observation on the effect of disinfection on gastroscope, contaminated by hepatitis B surface antigen (HBsAg) in the electrolyzed oxidizing water (EOW). METHODS: Contaminated gastric juice and serum was added to EOW for 1 minute. Positive control samples were treated with PBS instead of EOW in the same way. Gastroscopes used for hepatitis patients were immersed in the EOW for 1 minute after cleaning. Samples were collected before and after treatment. ELISA was used to test HBsAg. RESULTS: With mixed samples (average S/N = 42.16) of EOW, HBsAg became negative when diluted in 100 times. However, the HBsAg of positive control samples remained positive. After cleaning the gastroscope (average S/N = 5.99) immersed in EOW, HBsAg became negative. CONCLUSION: EOW was effective in destroying HBsAg which could be used for gastroscope disinfection. PMID- 11860844 TI - [Study on diarrhea disease and Escherichia coli strains harboring HPI pathogenicity island of Yersinia enterolitica in Shandong province]. AB - OBJECTIVE: It was found that some E. coli strains previously identified as entero SLTs-producing and invasive E. coli (ESIEC), harboring HPI pathogenicity island of Yersinia enterolitica. This research was designed to reveal prevalence, susceptible group and clinical features it caused. METHODS: All of the diarrhea patients were from the out-patient units of four hospitals in Shandong province from June to November of 1997. Stool specimen were collected before administration of antibiotics for isolation of enteric bacterial pathogen. Clinical symptoms were recorded at the same time. RESULTS: A total number of 449 enteric pathogenic bacteria strains were detected among stool samples from 671 patients, with isolation rates of Shigella species and diarrheagenic E. coli 25.48% and 15.05% respectively. When irp-2 and ipaB gene fragments were used as DNA probes, 42 of 176 un-identifiable strains were found irp-2(+) and ipaB(-), which were identified as HPI-harboring E. coli. Typical symptoms of the diarrhea caused by HPI-harboring E. coli were described as mild, including abdominal pain, shiver and vapidity. Temperature of the patients was usually normal. Over 6 times of bowel movements per day was frequently observed, of which most were unformed stools with mucous. CONCLUSION: HIP-harboring E. coli was an important diarrheagenic pathogen identified from 671 patients with diarrhea. Shigella was found to be the majority strain. PMID- 11860845 TI - [The study of countermeasures on measles control in infants]. AB - OBJECTIVE: To understand: a) measles antibody levels in purpura and their newborns, and the relation between them, b) maternal transferred measles antibody level of different age group infants, c) immunization rates of success in 6 and 8 month old infants, d) to study measles control countermeasures on infants. METHOD: To test measles IgG antibody levels of infant's blood using ELISA method. RESULTS: Antibody levels of measles in 58 pairs of mother and their newborns, 51 pairs were the same, while 7 pairs were different. Measles antibody levels in newborns was not high and reduced to 50% in 3 months, 20% in 6 months, and 15% in 8 months. The antibody levels in 6 and 8 month olds were low, nearly no protection to children. It was found that rates of immunization success and the antibody distribution were not statistically different between 6 and 8 months olds after measles vaccination. CONCLUSION: It was found that the measles antibody level in most matured women was low. Since mother's measles antibody level has close relation to their newborns, it seems that the newborn's antibody level can be improved by vaccination to matured women. In order to reduce morbidity of infant measles, the age of first vaccination of measles vaccine should be changed from 8 months old to 6 months old in some regions. PMID- 11860846 TI - [Analysis of causes of death and YPLL on residents in the industrial pollution area in Chongqing from 1991 to 1998]. AB - OBJECTIVE: To investigate the causes of death in residents living in the area of industrial pollution in Chongqing. METHODS: Mortality rate, sequence of causes of death, years of potential life lost (YPLL) and the valued years of potential life lost (VYPLL) were used to analyze causes of death in 1991 - 1998. Community not polluted by industry was chosen to serve as control. RESULTS: The annual average mortality rate of the residents was 7.34 per thousand (standard mortality rate 4.61 per thousand). The sequence of major causes of death was shown as below: malignant tumors (mortality rate 198.07/10(5), standard mortality rate 126.35/10(5)), cerebrovascular diseases (mortality rate 159.13/10(5), standard mortality rate 92.66/10(5)), respiratory system diseases (mortality rate 107.33/10(5), standard mortality rate 84.85/10(5)), cardiac diseases (mortality rate 95.36/10(5), standard mortality rate 59.37/10(5)) and accidental deaths (mortality rate 47.08/10(5), standard mortality rate 43.28/10(5)). Among malignant tumors, lung cancer took the lead with a mortality rate of 65.49/10(5) (standard mortality rate 45.27/10(5)). In both sequences of standard rates of YPLL and VYPLL for major causes of deaths, accidental death was always took the first place. CONCLUSION: In order to reduce mortality rate of the residents in the area, it is necessary to strengthen the administration of natural and social environment of the area. PMID- 11860848 TI - [Evaluation the pathological diagnostic values of koilocyte in condyloma acuminatum]. AB - OBJECTIVE: To evaluate the histological diagnostic values of koilocyte in histopathologic diagnosis of condyloma acuminatum. METHODS: Koilocyte was served as diagnostic criteria to scan 2 063 biopsies and compared the results with "goldan standard" of histopathologic diagnosis of condyloma acuminatum. RESULTS: Using koilocyte as diagnostic standard to diagnose condyloma acuminatum, the sensitivity and specificity were 77.93% and 95.26%, with positive predictive value 97.25%, negative predictive value 66.70%, positive likelihood rate 81.81, negative likelihood rate 23.17, under diagnostic rate 22.07%, misdiagnostic rate 4.74%. CONCLUSION: The koilocyte was proved to be one of classical histopathological morphologic features of human papillomavirus infection and other virus infection which served one of the most important histopathological markers for diagnosis of condyloma acuminatum, but not the only one. PMID- 11860847 TI - [Evaluation of p53, p21waf1, and PCNA in the diagnosis on lung cancer]. AB - OBJECTIVE: To evaluate the diagnostic value of p53, p21waf1 and PCNA on diagnosis of lung cancer. METHODS: One hundred and fourteen lung cancer patients and 89 cases with benign pulmonary diseases were studied on the expression of p53, p21waf1 and PCNA by immunohistochemical staining. The differences of p53, p21waf1 and PCNA expression status between these two groups were compared and the diagnostic value of these three markers was evaluated. RESULTS: (1) The positive rates of p53, p21waf1 and PCNA expression in lung cancer were 47.37%, 75.44% and 80.70%, all significantly higher than that in benign pulmonary diseases (P < 0.001), with odds ratio 39.15, 5.75, 6.76 respectively. (2) The positive likelihood ratio of p53 was 21.08, higher than that of p21waf1 and PCNA. Specificity and Youden index were also higher than those of p21waf1 and PCNA, but the sensitivity of p53 was low. (3) The sensitivity and specificity of p53 and PCNA parallel test were 89.47% and 60.67% respectively. CONCLUSIONS: (1) The expression of p53, p21waf1 and PCNA in lung cancer cases were all upregulated. These three markers all seemed to have strong links with lung cancer. (2) p53 was shown a good marker for lung cancer diagnosis. (3) The diagnostic value of p53 and PCNA parallel test was rather high. PMID- 11860849 TI - [Application of the interaction models between the polymorphism(s) of metabolic gene(s) and environmental exposure]. AB - OBJECTIVE: Taking GST M1 as an example to introduce analytic method of interaction models between the polymorphism(s) of metabolic gene(s) and environmental exposure in stomach cancer susceptibility. METHODS: Using community based case-control design, combined with molecular biological techniques (PCR) and multiple variables logistic regression models, we analyzed 112 intestinal types of stomach cancer cases with endoscopy and pathology diagnosis in the Yangzhong City Hospital during January 1997 and December 1998. A total of 675 controls were selected from persons who had no history of digestive system cancers. RESULTS: After adjustment of confounding variables with both GST M1 null genotype and history of ever tobacco smoking, the results showed a significant types of 4 gene-environment interaction. Interaction index (gamma) value was 3.38 and OR(eg) value was 8.40, suggesting that a super multiplicative interaction occurred. The results also showed that GST M1 null genotype had a high exposure gene effect interaction with tobacco smoking (pack year), while gamma values were 0.995, 2.085 and 2.157 respectively. A low exposure-gene effect interaction was found in GST M1 null genotype with the amount of (kg x year) alcohol consumption while gamma values were 1.01 and 0.97 respectively. CONCLUSION: Logistic regression model can be used to evaluate gene-environment interaction and dose response of exposure-gene effect. PMID- 11860850 TI - [A study on the epidemiology of Helicobacter pyroli infection among immigrants in Three Gorges area]. AB - OBJECTIVE: To investigate the epidemiologic features of Helicobacter pyroli (Hp) sero-prevalence rates among immigrants in Three Gorges area. METHODS: Enzyme labelled immuno-sorbant-assay was used to determine the Hp prevalence rate. A case-control study including 1 600 immigrants as cases and 1 600 non-immigrants as controls was carried out to explore the features of Hp infection in Three Gorges area. RESULTS: The prevalence rate of Hp in immigrant area was 67.50% higher than 53.19% that of Hp in non-immigrant (chi(2) = 13.40, P < 0.01). Rate of Hp infection was highest in infants than in children, increasing with age to clearly show a positive correlation between age and Hp prevalence rate (r = 0.62, P < 0.01). However, there was no marked sex difference in Hp infection rates in the immigrant area. Same results were also presented in the non-immigrant area. Hp prevalence rates were different which related to the stages of flooding. It is 89.17% in the area of first stage, while in second and third stages of flooding, the rates were 76.40% and 62.20% respectively. CONCLUSION: Preliminary result indicated that Hp infection rate of immigrant area was higher than that of non immigrant area. It appeared that living condition, environment, education condition might possibly have played important role in Hp prevalence. It is more important to prevent from infected children. Further study with respect to Hp prevalence rate was necessary when immigration is completed and the standard of living improved. PMID- 11860851 TI - [Analyses on the risk factors of Measles in Anhui province during 1998 and 1999]. AB - OBJECTIVE: To evaluate the measures targeting measles control. METHODS: Measles cases were studied serologically under AFP surveillance system. RESULTS: The incidence was 1.36/100 000 in 1999. The incidence of children under 5 was 3.65/100 000, with 84.00% and 87.49% reduction, as compared with the incidence in 1991 and 1997 before intensified vaccination campaigns in children aged 1 - 6. The indicators of surveillance showed an improvement in 1999 than that in 1998. CONCLUSION: Training for health staff at county, township, vallage levels played an important role in improving the sensibility and timeliness of measles surveillance system. Routine immunization is a key element in curbing measles outbreaks and reducing measles incidence. PMID- 11860852 TI - [Studies on epidemic feature of enteric adenoviruses infection in feces of infants with diarrhea in Shenzhen]. AB - OBJECTIVE: To estimate the enteric adenoviruses infection in feces of infants with diarrhea in Shenzhen. METHODS: All 195 fecal samples of infants were tested by PCR. Amplimers were analysed by Rsa I endonucleases and the type of enteric adenovirus identified. RESULTS: The positive rate of adenovirus DNA in fecal samples was 13.33% (26/195) while the prevalence rates of enteric adenovirus type 40, adenovirus type 41 and the other type of adenovirus were 2.56% (5/195), 8.21% (16/195) and 2.56% (5/195) respectively. Fever was the characteristic symptom of adenovirus infection. CONCLUSION: Enteric adenovirus type 40 and 41 were important pathogens which caused infant diarrhea in Shenzhen, with latter the majorty. PMID- 11860853 TI - [Study on hepatitis C virus genotyping in some parts of China]. AB - OBJECTIVE: To study the genotypes of hepatitis C virus in some parts of China. METHODS: Twenty-seven sera of plasma donors from Guan county of Hebei province, Zhoukou district of Henan province, and 36 patients with chronic hepatitis C from Beijing and Qingdao cities were tested for HCV RNA by Amplicor PCR kit. Thirty six of 38 HCV RNA positive sera were further genotyped, using INNO-LiP A HCV II kit. RESULTS: The HCV RNA positive rates of plasma donors and patients with chronic hepatitis C were 48.15% (13/27) and 69.44% (25/36) respectively. Genotyping data from 13 plasma donors and 23 patients with chronic hepatitis C showed that 29 (80.55%) were 1b, 5 (13.89%) were genotype 2, and 2 (5.56%) remained untyped. CONCLUSION: 1b seemed to be the prevalent HCV genotype in some parts of China. PMID- 11860854 TI - [Social economical status, behaviors and environment as the risk factors of tuberculosis in Chengdu China]. AB - OBJECTIVE: To study the risk factors of pulmonary tuberculosis in Chengdu, Southwest China. METHODS: A population-based case-control study was used which included 174 cases selected from 12 communities in Chengdu and 174 controls selected from registered population with normal chest radiograph. Cases were active TB patients which were matched for age, sex with controls, then interviewed by trained interviewers using a standardized questionnaire. RESULTS: Social economic status measured by education, occupation and income did not show obvions influence to TB; It found that the active smoking, passive smoking, type of cigarettes and alcohol consumption had no significanct effects on TB during logistic regression analysis. The study showed that a person who was smoking and also alcohol intake had a higher risk to get TB (OR = 6.12, 95% CI = 1.15 - 32.49). Significant association was showed in the Dose-Response Analysis (OR = 1.37, 95% CI = 1.30 - 2.30). Crowded living space (OR = 1.14, 95% CI = 1.05 - 1.25), degree of darkness (OR = 2.18, 95% CI = 1.11 - 4.27) and moisture (OR = 4.06, 95% CI = 2.25 - 7.33), poor sanitary (OR = 3.03, 95% CI = 1.22 - 4.44), airpollution of working environment, which were filled with dusts (OR = 2.35, 95% CI = 1.18 - 4.70) and chemical fumes (OR = 5.15, 95% CI = 1.44 - 18.40) were strongly associated with pulmonary tuberculosis. BMI also had strong relationship with TB (OR = 4.72, 95% CI = 2.68 - 8.33). CONCLUSION: Poor environment and exposure to dust and chemical fume under working condition, low BMI, smoking combining alcohol consumption were the risk factors of adult pulmonary tuberculosis in Chengdu, China. PMID- 11860855 TI - [Immunogenicity and safety of recombinant yeast-derived hepatitis B vaccine (YDV) on adults]. AB - OBJECTIVE: To study the immunogenicity and safety of recombinant yeast-derived hepatitis B vaccine (YDV) on adults. METHODS: One hundred twenty-four healthy teachers of 22 - 58 years old, were randomly selected in Beipiao city, Liaoning province. Their HBsAg, anti-HBs and anti-HBc were all negative and temperature was normal. They were vaccinated with 5 microgram/0.5 ml of YDV, which was made in Beijing Institute of Biologic Products, at 0, 1, 6 months. RESULTS: The positive rates of anti-HBs were 35.0%, 83.3%, 67.2% (P < 0.01) with the mean geometric titre (GMT) of anti-HBs 12.63 mIU/ml, 402.04 mIU/ml and 70.28 mIU/ml (P < 0.01) at 3th, 7th, 12th months respectively. The positive rates of anti-HBs and GMT of anti-HBs became the highest at 7th month, then began to decrease sharply. At 3th, 7th, 12th months the positive rates of anti-HBs and GMT of anti-HBs were higher on female compared to male, but only GMT of anti-HBs was significantly different at 3th month; the positive rates of anti-HBs in below 35 years old group was higher than 35 and over years old group, but it was significantly different only at 12th month. GMT of anti-HBs between the two groups was no obvious difference. Local or systematic addictive reactions were not found in all the subjects in 3 days after injection. CONCLUSION: YDV is effective and safety to adults. The duration of anti-HBs should be further observed. PMID- 11860856 TI - [Epidemiological observation on effect of Leptospiral outer membrane vaccine]. AB - OBJECTIVE: To study the safety and effect of Leptospiral outer membrane vaccine. METHODS: Eighty thousands dosages of Leptospiral outer membrane vaccine were vaccinated in Jingzhou and Shishou city Hubei prevince. Temperaure, side-effects such as Local edema with in 48 hours as well as the incidence of Leptospirsis within a year among those were vaccinated and unvaccinated were observed. RESULTS: (1) No any severe side-effect and abnormal reaction, was found, only 2 case suffered from slight fever and local edema which receded in 48 hrs. (2) Effects of Leptospiral outer membrane vaccine were as follows: 2 cases were attacked by lacterohaemorrhagiae in vaccination group and 47 cases in control group, so the protection rate 95.57% and confidence interval (CI) was 85.43% - 98.20%. Fifteen hebdomadis cases were found in control group. The protection rate of this vaccine reached 100.00%, CI 77.08% - 100.00%. CONCLUSION: Safty and protective effect were well showed when type of bacteria was concordant with that in vaccination district. PMID- 11860857 TI - [Immunogenicity and long-term persistence of anti-HAV in groups with different attenuated and inactived hepatitis A vaccine dosage]. AB - OBJECTIVE: This study was carried out to assess the long-term persistence of antibodies against hepatitis A in different vaccine dosage groups. METHODS: One hundred and sixty-six susceptibles aged form 6 to 8 years old were selected in Zhengding county, and were divided into 4 different dose groups. Each group received 10(6.17)TCID(50), 10(6.52)TCID(50), 10(6.83)TCID(50) of attenuated hepatitis A vaccine or 10(6.83)TCID(50) of the same vaccine but treated with 60 degrees C for 3 hours. All subjects were bled at month 3, 12, 24, 36 and 48. All serum specimens were tested for antibodies to HAV using Abbott-IMx mEIA reagent. Part of the tested negative serum specimens were retested by neutralization assay for 10(6.52)TCID(50) and 10(6.83)TCID(50) dosage groups at 4 years after vaccination. RESULTS: The seroconversion rate and GMT 3 months after vaccination in each dosage group showed a significant dose-response relationship. Antibody levels reached the peak at month 3 in each group but declined dramatically within one year and the decline slowed down thereafter. The serocoversion rate and GMT in 10(6.83)TCID(50) dose group 4 years after vaccination was significant higher than those of other dosage groups. Positive rates of neutralizing antibodies in B and C dosage group were 60.0% and 83.3% respectively. CONCLUSION: The immunogenicity and long-term persistence were related to the attenuated hepatitis A vaccine titer. The vaccine, whose titer was 10(6.83)TCID(50), had good immunogenicity and long-term persistence. Booster dose was not needed within short time after initial vaccination. PMID- 11860858 TI - [Cluster analysis study on the marshland of Schistosoma Japonicum using satellite TM image data in Peng Lake, Jiangxi province]. AB - OBJECTIVE: To create a category land cover map of the marshland region of Schistosoma Japonicum using satellite TM data. METHODS: TM satellite images from Peng Lake of Jiangxi province were applied to the Cluster analysis. Then the resulting clusters were identified and reclassified by undertaking site visits. RESULTS: Eight land cover classes were generated, including Carexspp Zone that was the snail habitat place. CONCLUSION: Cluster analysis, which is a technique for the interpretation of remotely sensed imagery, could contribute to the study on the distribution of snail habitats and become a new tool for the epidemiological ecology study. PMID- 11860859 TI - [Epidemiological study on the influence of pig-derived Ascaris to the transmission of human ascariasis]. AB - OBJECTIVE: An experimental epidemiological study was designed to investigate possible relations of pig-derived Ascaris to the transmission of human ascariasis which might be caused by cross-infection between the pig and humans. METHODS: In Xinjian County, Jiangxi province, two adjacent villages, Laozhi and Panzhi, with similar baseline of ascariasis in pig and human populations were selected as the study sites with a one-year longitudinal epidemiological follow-up study. People in the two villages received mass chemotherapy. Pigs received mass chemotherapy only in Panzhi village and was twice repeated with two months interval, while pigs in Laozhi village were not treated at all. In the following year after treatment on humans, ascariasis among villagers was studied cross-sectionally five times with Kato-katz technique. RESULTS: No significant difference of the re infection patterns of human ascariasis was found between the two villages in terms of prevalence and intensity during the following year in all the five surveys (chi(2) < 0.658, F < 1.658 and P > 0.1). CONCLUSION: The results suggested that pig-derived Ascaris were mainly transmitted among pigs but had no important role on the transmission of human ascariasis. The results were in good accordance with the findings through a recent molecular genetic research on the local human-and pig-derived Ascaris worms. Possible confounding factors were discussed. PMID- 11860860 TI - [Meta-analysis of risk factors on lung cancer in non-smoking Chinese female]. AB - OBJECTIVE: To evaluate the risk factors of lung cancer in non-smoking Chinese women. METHODS: The results of 7 case-control studies from 1990 to 1999 were analyzed by Meta-analysis method in this study. The cumulative cases and controls were 1 115 and 1 520, respectively. Der Simonian and Nan Laid model were applied to process data. RESULTS: The pools OR values of family history of lung cancer, personal history of non-neoplastic lung disease, cooking oil fume pollution, coal pollution and exposure to environmental tobacco smoking (ETS) were 2.87, 2.79, 2.52, 1.42 and 1.64 respectively. CONCLUSION: Family history of lung cancer and personal history of non-neoplastic lung disease might serve as the most important risk factors of female lung cancer in China. There appeared also significantly positive relationship between female lung cancer and the degree of cooking oil fume pollution or pollution of coal burning. The effect on lung cancer of exposure to ETS was uncertain in this study which calls for further investigation. Bias and confounding factors were also discussed in this study. PMID- 11860861 TI - [Study on the epidemiology and factors influencing the behavior of school children in Taiyuan]. AB - OBJECTIVE: To study the epidemiological features and factors influencing behavior problems of school children in Taiyuan. METHODS: One thousand five hundred and ninety-seven school children aged 6 - 11 years selected from three schools differently located in three city proper were measured by Achenbach Child Behavior Checklist. RESULTS: Pervaleuce rate of children having behavior problems averaged 12.34%, with 12.40% in boys and 12.27% in girls. If was found higher in Hexi than in other two schools. The total scores of CBCL hyperkinesia and discipline violation among boys were higher than that of girls. The score including depression, somatic problem, and social withdraw of girls was found higher than that of boys. The result of multiple regression indicated that factors as interest in study, easy rearing, and good living condition when mother was in pregnancy were possibly benefit to behavior development in children, while beating and scolding children, indifference toward children, and particular preference of diet seemed to be the main risk factors affecting children's behavior. CONCLUSION: There appeared different rates of childrens behavior problems among pupils. The scores of CBCL were different between boys and girls. It is essential to adopt comprehensive measures for the prevention of behavior problems in children. PMID- 11860862 TI - [Study on the prevalence rate of primary osteopenia in the middle-aged and the elderly population in some parts of China]. AB - OBJECTIVE: To understand the distribution of prevalence rate of primary osteopenia (POPA) in the middle-aged and the elderly in some parts of China. METHODS: Bone mineral density (BMD) was measured and questionnaire was administered to 5 600 people aged above 40 in five administrative areas in China, through a stritified-multi-steps-cluster sampling method. RESULTS: The total prevalence rate of primary osteopenia in some parts of China was 15.8%, according to the most significant parts of the rate of loss. Of them, the prevalence rate of males was 20.0% compared to 12.4% in females (P < 0.01); There were also differences on POPA prevalence among provinces, age groups and areas between urban and rural (P < 0.01). CONCLUSION: Although prevention and treatment of primary osteopenia were important strategy in males however it should not be ignored in females as well. PMID- 11860863 TI - [A study on the epidemiological characteristics of obesity in Chinese Adults]. AB - OBJECTIVE: To elucidate the epidemiological characteristics of obesity in Chinese adults and to provide scientific basis for public health policy-making, as well as for prevention and control of obesity among Chinese. METHODS: Using the data from diabetes mellitus, epidemiological studies carried out in 11 provinces/autonomous regions/municipalities of China from July 1995 to June 1997, including 42 751 subjects aged 20 - 74 years who had lived in the communities for no less than 5 years. Diagnostic criteria was based on the recommendation from WHO. Epidemiological characteristics of overweight and obesity were analysed. RESULTS: The prevalence rate of overweight and obesity among Chinese adults were 21.51% and 2.92% respectively. The age-standardized prevalence rates of overweight and obesity standardized by Chinese population aged 20 - 74 years were 18.28% and 2.48% respectively in 1990, when standardized by the standard population of world in 1992, the prevalence rates were 18.61% and 2.49% respectively. The prevalence of overweight and obesity in female (21.71% and 3.73% respectively) were significantly higher in male (21.25% and 2.11% respectively), the prevalence rates of obesity in male and in female increased with the increase of age, with the lowest prevalence in 20 year old group (1.23% and 0.96% respectively), the highest were 60 year old group (3.30% and 6.91% respectively). Among 11 provinces, the highest prevalence rates of overweight and obesity were in Beijing (34.75% and 5.14% respectively) of males, and in Shandong (34.56% and 9.36% respectively) of females. The prevalence rates of obesity were significantly higher in northern part of the country (2.64% in males and 4.85% in females) then in the south (1.36% in males and 2.15% in females), as well as significantly higher in urban (2.65% in male and 4.21% in female) than in rural (0.88% in male and 2.80% in female). CONCLUSION: Overweight and obesity had become prevalent in Chinese adults, more seen in the north than in the south, more in females than in males, and in older age than in younger, suggesting an earlier research and practice of prevention and control for overweight and obesity need to be implemented in China. PMID- 11860864 TI - [A matched case-control study on direct economic costs of four kinds of nosocomial infections]. AB - OBJECTIVE: To evaluate the direct economic costs of four major kinds of nosocomial infections such as nosocomial pneumonia, urinary tract infection, deep surgical wound infection and intracranial infection. METHODS: A matched case control study was conducted among 95 pairs of hospitalized patients to investigate the direct economic costs of nosocomial infections. RESULTS: The average medical cost for patients in the case group was 38 741 RMB Yuan, 20 365 Yuan more than that in the control group patients (P < 0.0001). The largest part of extra costs was found in drug expenses, taking up 62.07%, while other extra costs such as non-drug therapies, blood transmission and laboratory examinations, accounted for 10.32%, 7.40% and 5.72% respectively. Extra economic burden caused by nosocomial infections varied in different degrees with different departments, with the heaviest found in the department of tumourology and hematology at an average extra cost of 34 944 Yuan. The average extra charges in case group caused by nosocomial pneumonia, urinary tract infection, deep surgical wound infection and intracranial infection were 31 940 Yuan, 7 436 Yuan, 17 332 Yuan and 17 349 Yuan respectively. The average length of hospital stay was 50.57 days in case group and 25.22 days in control group, showing a 25.53 extra days (P < 0.0001) of hospitalization due to nosocomial infections. CONCLUSION: Nosocomial infections had significantly added to the economic burden of managing the underlying diseases, led to prolonged hospitalization of the patients and lowered the turnover rate of hospital bed. There is a need to call for better infection control program, which would bring tremendous social and economic profits. PMID- 11860865 TI - [Investigations on Human ehrlichia infectious people in Daxingan Mountains]. AB - OBJECTIVE: In order to investigate the prevalence of Human ehrlichia infectious people in Daxingan Mountains. METHODS: Using PCR detection, followed by sequence analysis to detect EC and Human granulocytic ehrlichia from people in Daxingan Mountains applying the 16S rRNA gene primers of the two agents. RESULTS: Specific 16S rRNA gene sequences of EC and HGE agent were amplified from human blood specimens of tick-bitted patients and woodsmen. The detected HGE agent 16S rRNA gene (1 433 bp) amplified from a blood specimen of a tick-bitted patient differed in only one base compared with that of an American HGE agent strain. CONCLUSION: It was considered that Human ehrlichia infectious people did exist in Daxingan Mountains. PMID- 11860866 TI - [Cost-effectiveness analysis of hepatitis B vaccination in People's Liberation Army]. AB - OBJECTIVE: To select an optimal vaccination approach and provide basis for decision-making on the control of hepatitis B infection in PLA. METHODS: Decision trees were constructed. The yearly new infection rate in susceptible cohort, using a catalytic model, was estimated. Disability-adjusted life year (DALY) was used to assess the effectiveness. Incremental cost-effectiveness ratio (CER) was then calculated. RESULTS: Compared with no vaccination, DALY of immediate vaccination and vaccination after screening were 61.89 and 57.18 with CER 392.70 and 251.90 respectively. After weighting with actual age construction of PLA population, the CERs of vaccination after screening and immediate vaccination were 251.90 and 392.70 respectively. CONCLUSION: The results indicated that screening followed by vaccination was superior to immediate vaccination approach. The younger the vaccination was carried out, the more cost-effective was shown by vaccination. PMID- 11860868 TI - [Study on the optimal perinatal health care and child health care surveillance systems]. AB - OBJECTIVES: To study the establish an optimal perinatal health care (PHC) surveillance system and an child health care (CHC) surveillance system in China, that can provide better health care services for women and children. METHODS: All women receiving marital check-ups are issued PHC booklets and then their menstrual dates are recorded regularly. If a woman is confirmed to be pregnant, she will receive her first prenatal exam. Subsequent prenatal exams and visits will be followed and recorded accordingly. CHC data will be collected regularly during the provision of routine child health care services. RESULTS: Surveillance systems have been set up in 32 project counties/cities in China, comprising more than 20 million people. The surveillance systems have smoothly been conducted for eight years since 1993. CONCLUSION: The surveillance systems have been proved to be adaptable under China's context. Both PHC and CHC systems seemed to be helpful in improving the health care services as well as the health condition of mothers and children in the areas. PMID- 11860867 TI - [The establishment of an electronic reproductive health surveillance system (ERHSS)]. AB - OBJECTIVE: To establish an electronic reproductive health surveillance system (ERHSS) and a population-based, computerized monitoring and management information system. METHODS: The ERHSS was designed based on the existing perinatal health care (PHC) surveillance, birth defects (BD) surveillance and child health care (CHC) surveillance systems. Advanced computing and network techniques were adopted in designing and developing this system. The use of information technology (IT) and its products has become popular in public health surveillance. RESULTS: The ERHSS was developed by National Center for Maternal and Infant Health (NCMIH) and Beijing InfoUnit Technological Corporation. It consists of four main components: (1) The establishment of electronic data collection system, including the electronic forms and the screening system for high risk factors; (2) The established electronic data transmission system; (3) The established electronic information reporting system; (4) The established back up system, including card making system, ID search engine and maintenance, etc. CONCLUSION: It is the first time this population-based electronic surveillance system being developed in China. It has many features regarding accuracy, security, smooth operation, optimal performance of real-time data processing, etc. The ERHSS has been set up in 22 counties/cities in China since 1999. PMID- 11860869 TI - [A population-based surveillance system on birth defects and its application]. AB - OBJECTIVE: To describe a unique surveillance system on birth defects in the People's Republic of China. METHODS: The system was instituted in 1992 as a component for the evaluation on the effectiveness of a community intervention program using periconceptional folic acid supplementation to prevent neural tube defects and currently carries out surveys on a birth cohort of approximately 150 000 infants per year. Information was collected in the form of detailed written descriptions by local health care providers and photographs of affected infants. The system allowed for detection of birth defects at the local level with later definitive classification and coding; however, information was limited to those structural anomalies which were visible on physical examination. RESULTS: This surveillance system on birth defects provides an extensive database of infants with major and minor external structural anomalies. From 1993 to 1996, more than one million births were monitored, with more than 10 000 birth defects and 30 000 photos submitted. Among them 5 000 were identified with major birth defects. Neural tube defects, oral clefts and extremity abnormalities were the first few defects with most cases. Data were used in birth defect surveillance reports and scientific papers. CONCLUSIONS: This system can be utilized for studies on etiology, descriptive epidemiology, and identification of unusual trends which is unique with multiple advantages in the People's Republic of China. PMID- 11860870 TI - [A study on the risk factors of congenital urogenital malformations]. AB - OBJECTIVE: To explore risk factors for congenital urogenital malformations. METHODS: A hospital-based category matching case-control study was performed to investigate the characteristics of 95 cases with congenital urogenital malformations and 160 controls. Parents were interviewed for their characteristics before and during conception. Unconditional logistic regression analysis was used for single factor and multivariate analysis to estimate odds ratios and their 95% confidence intervals. RESULTS: There were 71 male infants in the overall congenital urogenital malformations (Ratio = 74.4%, Chi(2) = 74.863, P = 0.001) identified. Statistically significant associations were calculated for the pregnant women suffering from cold during first and/or second trimester (OR = 7.034, 95% CI = 3.488 - 14.187), for a low birthweight delivery (OR = 4.075, 95% CI = 1.774 - 9.362), for maternal times of pregnancy >/= 2 (OR = 3.133, 95% CI = 1.514 - 6.483) and exposure to occupational detrimental chemicals before pregnancy (OR = 10.496, 95% CI = 1.053 - 104.651). CONCLUSIONS: The risk factors of congenital urogenital malformations were suffering from cold during first and/or second trimester, male infants, low birthweight delivery, times of pregnancy >/= 2 and exposure to occupational detrimental chemicals before pregnancy. PMID- 11860871 TI - [Case-control study on sexual coercion and related risk factors in China]. AB - OBJECTIVE: To find out the determinants of sexual coercion among adolescent abortion seekers in China. METHODS: Case-control study was adopted. Women seeking for abortion and ever having experienced sexual coercion were taken as case group and those who had never experienced sexual coercion as control group. Case control study was carried out in 11 large hospitals in Beijing city from January 2000 to April 2000. RESULTS: The number of subjects in case group was 512, comparing to 517 in the control group. The mean age of subjects was 20.25 years with monthly income 881.2 RMB Yuan. The difference between case group and control group in income, occupation and age showed no statistical significance. The results of the study indicated that the factors which were more likely to be related to sexual coercion include lower educational level (chi(2) = 15.27, P < 0.01, OR: 1.71), not living with parents (chi(2) = 10.18, P < 0.01, OR: 1.50), mobile nature (chi(2) = 20.60, P < 0.01, OR: 1.78), experienced battery by partner (chi(2) = 6.79, P < 0.01, OR: 2.72), abused by her partner (chi(2) = 4.22, P < 0.05, OR: 1.71) multi-partners (chi(2) = 19.08, P < 0.01, OR: 2.17), sex after being drunk (chi(2) = 34.22, P < 0.01, OR: 5.04), and larger gaps of in age between male and female partners (chi(2) = 11.04, P < 0.01, OR: 2.17). CONCLUSION: Factors as, poor-educated, not living with parents, floating population, multi-partners, the inequality between male and female were more likely to increase the risk of sexual coercion. PMID- 11860872 TI - [Investigation on an outbreak of Legionnaires' disease caused by Lboz in a suburb of Beijing]. AB - OBJECTIVE: To demonstrate the infectious rates and incidence rates through epidemiological investigation. METHODS: Serum samples from 203 new soldiers were detected from Lboz by TAT. RESULTS: Infectious rates was 33.00% (67/203), and the incidence rates was 8.87% (18/203) with 2 cases of pneumonic type and 16 cases of pontiac fever type. CONCLUSION: The results suggested that an outbreak of Legionnaires' desease caused by Legionella bozemanii (Lboz) took place in a suburb of Beijing. However the report of the outbreak caused by Lboz had not been found in related references. PMID- 11860873 TI - [Seroepidemiological analysis of characteristic of hepatitis B virus in children after vaccination in Ningbo]. AB - OBJECTIVE: To understand the immunity of children in Ningbo after hepatitis B vaccination for seven years and to explore the best time for booster. METHODS: One thousand nine hundred and twenty serum samples from children born in 1982 - 1995 were tested for anti-HBs and HBsAg, using RIA. RESULTS: (1) HBsAg positive rate (2.66%) was lower in vaccinated children born in 1992 - 1995 than those unvaccinated (10.92%) born in 1982 - 1995. HBsAg positive rate had a drop of 75.71% (chi(2) = 22.91, P < 0.0001). (2) Anti-HBs positive rates (65.95%) in vaccinated children was much higher than that in unvaccinated children (41.03%) (chi(2) = 113.67, P < 0.0001). (3) It also showed that children's positive rate of HBsAg increased and anti-HBs dropped after 5 - 7 years of vaccination. CONCLUSION: After immunization of hepatitis B vaccine for 7 years, high immunity and low infection rate of HB in children it appeared in Ningbo. However it is necessary to further study the reason causing the increase of HBsAg positive rate and/or deerease of anti-HBs positive rate, and the suitable time frame of boostering, after 4 or 5 years of primary vaccination. PMID- 11860874 TI - [Epidemiological effects of live attenuated hepatitis A vaccine (H(2)-strain): results of A 10-year observation]. AB - OBJECTIVE: To assess the long-term epidemiological effects after a mass vaccination program, using live attenuated hepatitis A vaccine (H(2)-strain). METHODS: Shengsi county and Jiaojiang city in Zhejiang province were chosen where high incidence rate of hepatitis A was observed. Mass vaccination program of H(2) strain vaccine was carried out on 1 - 15 year olds. Cohort studies included two groups were carried out in Shengsi. Cases reported program on hepatitis A was under the national standard on clinical diagnosis and partly through laboratory monitor of anti-HAV IgM and IgG, ELISA and Abbott EIA. Population size was estimated by the yearly reports of census bureau of the county and the city. Serum tests including neutralization test, immune-recall responses, and anti-HAV IgG were performed 10 years after the initial immunization. RESULTS: The morbidity of hepatitis A was significantly different between vaccinated groups and non-vaccinated groups (P < 0.01) in the cohort study. The protective efficacy in the site was 100 percent. There was a significantly negative correlation of regression between vaccine coverage and hepatitis A morbidity. In both sites, hepatitis A incidence rate showed decrease of 94.5% and 90.3% in whole population, with 96.9% and 97.9% in 1 - 15 year olds respectively. Neutralization test showed that 63.8% of the serum negative to ELISA after vaccination could still neutralize certain HAV. Immune-recall response after the booster was seen in vaccine group with 80.2% positive rate of anti HAV-IgG, 10 years after immunization among vaccinees in both sites. Not a single hepatitis A case was identified. CONCLUSION: Live attenuated hepatitis A vaccine seemed to have obviously long-term epidemiological effects. PMID- 11860875 TI - [Analysis on the epidemiology of syphilis in Guangdong province from 1995 - 1999 and the strategy for syphilis control]. AB - OBJECTIVE: To explore the epidemiologic characteristics of syphilis in Guangdong province based on the data from 1995 - 1999 in the province and to provide reference for further control of the disease. METHODS: Registered cases of syphilis were collected in every city/county and the data was analyzed epidemiologically. RESULTS: The incidence rate of syphilis in Guangdong province increased rapidly the past five years, went up from 1.81 per 100 thousand residents in 1995 to 15.17 per 100 thousand residents in 1999 with an average increase as high as 133.37%. The proportion of syphilis in the eight-reportable STD also increased annually with an average annual increase rate of 125.40%. Among the cases, primary cases were dominant accounting for 91.02%. Most patients were males with 20 - 29 year olds accounted for the majority. As for education level, the majority of the patients were middle school graduates. The most popular way of transmission was through extra-marrital sexual contact which accounted for 77.31%. Syphilis patients were mainly seen in coastal cities such as Guangzhou and Shenzhen. CONCLUSION: Based on the epidemiologic feature of syphilis in Guangdong, following measures should be taken in order to control the disease: to enhance the STD knowledge through education, to promote the widely use of condoms and to standardize the diagnosis and treatment of syphilis. PMID- 11860876 TI - [Trend and characteristics of HIV/AIDS epidemic in Guangdong province, 1986 - 1999]. AB - OBJECTIVE: To analyze the epidemic characteristics and risk factors of HIV/AIDS in Guangdong province and to search for measure of control. METHODS: Data from routine, sentinel, behavior and laboratory surveillance programs were collected and evaluated. RESULTS: Since the first HIV case in IDUs was found by the end of 1996, the number of HIV/AIDS has increased by 86.2% yearly and the proportion of cases infected through IDU increased rapidly from 1.4% before 1996 to 91.0% in 1999. HIV-1 C subtype in IDUs accounted for 61.3%. CONCLUSIONS: Number of HIV/AIDS cases has increased rapidly in recent years in Guangdong. The major transmission route was through IDU. It was important to prevent HIV/AIDS spreading from IDUs to sex workers then to the general population through behavioral surveillance and intervention. PMID- 11860877 TI - [Dynamic analysis of incidence and mortality of stroke and the risk factors in the communities in Shanghai, in the 1990's]. AB - OBJECTIVE: To understand the trend of incidence, mortality of stroke and the risk factors at the community level in the 1990's. METHODS: To establish a network of disease surveillance. Both incidence and mortality of stroke were surveyed in the 35 - 74 age group during 1991 - 1999 using the method of WHO MONICA. By cluster sampling in 1992 and 1998, 500 individuals aged 35 - 59 from each female and male group were investigated respectively on the risk factor of stroke. RESULTS: The incidence of stroke was averaged 168.2 per 100 000 in the group aged 35 - 79 during 1991 to 1999 with 194.5 and 144.9 pre 100 000 respectively for the male and female group of this age. The incidence of stroke in the male group was higher than that in the female group, also with a faster speed of increase in the male group. The overall mortality of stroke was averaged 79.2 per 100 000 with 87.2 and 71.4 per 100 000 in male and female groups respectively of this age group. The incidence of stroke increased with age in both male and female groups. The average age of onset had been 66.1 year olds during 1996 - 1999, 3.5 years older than that age groups during 1991 - 1995, but did not show much difference between male group and female group. CHO, Blood Pressure, BMI, Prevalence of Hypertension, TC/HDLC, WHR, Prevalence of High-CHO, Prevalence of Low-HDL all had a tendency of increase in both male and female groups. The rates of smoking cigarettes and drinking alcohol both decreased, especially in the male group. CONCLUSION: The incidence of stroke and some relative risk factors seemed to increase in the 1990's, but the mortality of stroke had a decrease in the same period in the communities in Shanghai. PMID- 11860878 TI - [Association between apolipoprotein E polymorphism and Alzheimer's disease: A population-based study in Guangzhou, China]. AB - OBJECTIVE: To investigate the distribution of apolipoprotein E (apoE) genotype of patients with Alzheimer's disease (AD) in the population aged 75 and older in Guangzhou and to determine whether there was an association between AD and apoE epsilon4 allele. METHODS: A systematic cluster sampling design, two-stage screening and field evaluations were used in a cross-sectional population-based study. Three thousand eight handred and twenty-five subjects aged 75 and above in Liwan district of Guangzhou were interviewed. Blood samples were collected from 110 cases and 518 normal controls. The criteria of DSM-III-R and NINCDS-ADRDA were used for clinical diagnosis of probable AD. Polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) were used to determine apoE genotypes. RESULTS: There were 190 cases diagnosed as sporadic AD from this sample. The frequencies of the three apoE alleles were detected as follows: epsilon2 = 0.100, epsilon3 = 0.832, epsilon4 = 0.068 in AD patients; epsilon2 = 0.092, epsilon3 = 0.843, epsilon4 = 0.065 in normal controls. The frequencies of apoE genotype and allele in the AD patients were not significantly different from that in normal subjects (P > 0.05). CONCLUSION: No association between sporadic AD and apoE epsilon4 allele was found among the population aged 75 and above in Liwan district of Guangzhou. PMID- 11860879 TI - [Study on the quality of life among the elderly in the rural and pastoral districts in Baotou city]. AB - OBJECTIVE: In order to explore the situation of QOL of the elderly in the rural and pastoral districts in Baotou city, to compare the differences of QOL between the two districts, and to find out the major factors influencing the QOL of the elderly. METHODS: SF-36, recommended by WHO was adopted to study the QOL of 1 421 old people in the rural and pastoral districts in Baotou city. RESULTS: (1) The average weighted score based on the SF-36 was 64.16 +/- 18.76 in the rural area district and 73.94 +/- 17.48 in the pastoral district (F = 137.8, P < 0.01). The level of QOL of the elderly in the pastoral district was significantly higher than that in the rural district. (2) Results from the multiple liner regression analysis showed that the main factors influencing the QOL of the elderly were chronic disease, age, hospitalization, education and gender in the rural district, but chronic disease, age, hospitalization, income and marital status in the pastoral district. CONCLUSION: Population with older age, low education, no spouse and low income should be viewed as the priority of attention and their special difficulties fully considered when setting up the policy of health care and social security. PMID- 11860880 TI - [A study on the risk factors of cerebral infarction as complication of type 2 diabetes mellitu patients]. AB - OBJECTIVE: Cerebral infarction, a common acute cereovascular disease, is one of the main complications of type 2 diabetes mellitus (DM). This paper is to explore the risk factors of cerebral infarction among type 2 DM patients. METHODS: A pair matched case-control study was carried out. Data was analyzed with conditional logistic regression. RESULTS: One hundred and ten type 2 DM patients with cerebral infarction and controls were selected respectively. Results showed that higher educational level and physical exercises lowered the risk of cerebral infarction among type 2 DM patients. Higher HDL-C and blood sugar under control could also lower the risk of developing cerebral infarction among type 2 DM patients. The above factors seemed to have protective functions. Excessive salt intake and snoring increased the risks of cerebral infarction development among type 2 DM patients. Hypertension served as an independent risk factor while atrial fibrillation and hyperinsulinemia were important risk factors. High LDL-C, Apo-A, Lp(a), Fg increased the risk of cerebral infarction among type 2 DM patients. The study did not discover the relationship of smoking, drinking, oral contraception, menstrual and obstetrical history, TC, TG, Apo-A and cerebral infarction among type 2 DM patients. CONCLUSION: To prevent cerebral infarction among type 2 DM, measures need to be taken as control of salt intake; eating more vegetables; regular physical exercises; control of blood sugar, blood pressure and Hyperinsulinemia. PMID- 11860882 TI - [Study on the risk factors of psoriasis]. AB - OBJECTIVE: To investigate psoriasis patients with family history and non-family history on their age and season of onset, disease severity and predisposing factors. METHODS: Two hundred and twenty psoriasis patients with family history and 547 psoriasis patients without, together with 647 normal controls have been investigated through questionnaires. EPI info 6.0 method was applied for data analysis. RESULTS: (1) In 220 psoriasis patients with family histories, morbidity rates of the first and second-degree relatives were 16.98% and 2.7% respectively. (2) Both mean age and age of onset were younger in females than that in males with family histories (t = 1.715 0, P < 0.05). The mean age of onset was younger in males with family histories than those without. However, there was no significant difference found in females (t = 0.739 2, P > 0.05). The mean age of onset seemed to be younger in females with family histories than that without, but with no significant statistical difference. (3) There were no significant differences in the age and season of onset, disease severity, and predisposing factors between groups with family histories and without. However, environment factors seemed to influence not only psoriasis patients with family histories but also those without, which showed significant difference (chi(2) = 0.23, P < 0.05). CONCLUSIONS: There was no significant difference in the following aspects as season of onset, disease severity and predisposing factors between the ones with family history and those without, except the age of onset and age of average. The results showed that genetic factor might play an important role in psoriasis patients with family history. It also demonstrated that psoriasis is a multifactor hereditary disease. PMID- 11860883 TI - [A survey on contraceptive knowledge, attitude and behavior among never-married young women who are seeking pregnancy termination in Beijing]. AB - OBJECTIVE: To identify contraceptive knowledge, attitude, behavior and its determinants among never-married young women who have unwanted pregnancy in Beijing. METHODS: A cross-sectional study under adoption of Lawrence "Procede Proceed" model, was conducted in China in 1999. Three hundred and six unmarried young women aged 18 to 24, requesting for pregnancy termination were interviewed in person. RESULTS: Findings of this study indicated that only one-eighth (13%) of the young women insisted on contraceptive use every time, and almost an equal proportion (26.4%, 26.8% respectively) occasionally or never using contraceptives. Among 224 women who ever used contraceptives during the past 12 months, condom (49%) ranked the first place followed by withdrawal (27.7%) and the rhythm method (15.6%). One of the most important reasons, cited by 73 percent of women who had never used contraceptives, was that they were not aware of the risk of pregnancy when engaging in sexual activities. The results of logistic regression analysis revealed that knowledge about contraception, boyfriend's approval of contraceptive use, perceived the risk of getting pregnancy, perceived availability of contraceptive service and discussion of contraception with boyfriend were important indicators of young women's behavior on contraceptive use. CONCLUSION: These results indicated an urgent need to develop sex education about contraception among young women and men, through enhancing perception about risk of unwanted pregnancy and complications of induced abortion among young women, and to promote men's co-operation and participation in contraceptive use as well as strengthening communication on contraception between young women and their partners. PMID- 11860885 TI - [A simplified and quantitative method for culturing and isolating Legionella from air condition cooling waters]. AB - OBJECTIVE: To establish simplified technique to investigate the distribution of Legionella from air condition cooling waters in a district in Shanghai. METHODS: Using simplified method to make GVPC agar plates; using candle jar to replace CO(2) incubator; initial identification was only relied on colony appearance but no growth on the blood agar plate; and a simple formula was used to quantify the Legionella in waters. RESULTS: From July 1997 to June 1999 in Luwan District Shanghai, the simplified methods were used to culture and isolate 185 Legionella strains from 371 water samples (47 fixed spots of hotels, markets, resident buildings, hospitals, subway stations, cinemas and stadiums), with a detection rate of 49.87% with highest at the subway stations (69.36%), which were significantly higher than repeated in the literature in this country and abroad. CONCLUSIONS: Simplified methods were established for culturing, isolating and quantifying Legionella. These methods had been used for 3 years to have got a high Legionella detection rate from the cooling waters. The high detection rate and quantity of Legionella in cooling waters of subway stations call for more attention to the fact that Legionellosis might exist in these waters. PMID- 11860881 TI - [A study on nosocomial infection among inpatients in Beijing Hospital for elderly]. AB - OBJECTIVE: To investigate the nosocomial infection among inpatients. METHODS: A retrospective study on 12 418 inpatients was undertaken from January to December in Beijing Hospital in 2000. RESULTS: Three hundred and seventy-eight cases due to nosocomial infection were identified with a rate of 3.04%. There existed significant differences with rates of different sex and ages. The infection rate in males was higher than in females. The older the age, the higher the rates of infection was noticed. Nosocomial infections occurred mainly in the lower respiratory tract (35.99%), upper respiratory tract (22.46%), urinary tract (17.63%) and gastrointestinal tract (10.39%). The main pathogenic organisms of the nosocomial infection were Candida albicans (15.78%), Pandomonos (13.15%) and Enterococcus (11.84%) with gram negative bacilli (46.03%), gram positive coccus (23.68%) and fungus (15.78%). The overall death rate of nosocomial infection was 14.29%. CONCLUSIONS: Nosocomial infections were closely correlated with sex, age, department of wards and invasive operation etc. Prevention and proper treatment of nosocomial infection should be strengthened. PMID- 11860884 TI - [Stratified 'capture-recaptured' methodology in the estimation on the number of cancer incidence cases]. AB - OBJECTIVE: To develop a new method in estimating the number of cancer incidence in Yuexiu district in Guangzhou. METHODS: Data of cancer incidence was collected through hospital records and death certificates to develop a stratified capture recaptured model. Related procedures for the parameter estimation were given and implemented on SAS language. RESULTS: Estimated numbers of both male and female cancer incidence in Yuexiu district were 610 and 520 respectively. The estimated miss-reporting rates on both male and female cancer incidence from hospital records and death certificates were 8.5% and 5.4% respectively. CONCLUSION: The result of simulation studies showed that stratified "capture-recaptured" model proposed in this paper was reliable for the estimation of the number of cancer incidence cases. PMID- 11860888 TI - [Preliminary study on relationship between antibody against heat stress protein 70 and hypertension]. AB - OBJECTIVE: To preliminarily explore the relationship between main antibody against heat stress protein 70 (anti-HSP70) and hypertension. METHODS: Serum anti HSP70 was determined with Western blotting technique in hypertensive and normotensive groups. Logistic regression model was used to analyze the relationship between anti-HSP70 and hypertension. RESULTS: It was found that level of antibody against the main heat shock protein 70 was significantly higher in patients with hypertension (based on criteria of 160/95 mm Hg, 1 mm Hg = 0.133 kPa) than in controls, after adjustment for the generally recognized risk factors for hypertension (such as age, sex, length of employment, body mass index, job, et al.). There still was stronger association between anti-HSP70 and hypertension (based on criteria of 140/90 mm Hg), but its strength of association was weaker than the former. CONCLUSIONS: Anti-HSP70 may potentially serve as a serum biomarker to assess hypertension, or anti-HSP70 itself may be involved in the development of hypertension. PMID- 11860886 TI - [Analysis for the titer of plasma antibody to heat stress protein 70 in workers exposed to benzene]. AB - OBJECTIVE: To explore the titer of plasma antibody against heat stress protein 70 (HSP70) in workers exposed to benzene and its possible relationship to their health. METHODS: Forty-two workers exposed to more than 40 mg/m(3) of benzene were selected as a high exposure group, 50 of workers exposed to less than 40 mg/m(3) as a low exposure group, and 42 unexposed as a control group. Their plasma antibody against HSP70 was determined with enzyme-linked immunosorbent assay (ELISA). Analysis was conducted to study the relationship between it and other factors. RESULTS: Proportion of plasma positive antibody to HSP70 with a titer of 1:20 was significantly higher in high exposure group (26.2%) than that in controls (9.5%). Moreover, prevalence of adverse symptoms and positive signs, and level of lipid peroxide were all significantly higher in high exposure group with positive antibody to HSP70 than those with negative antibody. CONCLUSION: It was concluded that the titer of plasma antibody to HSP70 had close relation with the health of workers exposed to benzene. PMID- 11860887 TI - [A study on relationship between plasma heat stress protein 70 and cytokine in patients with heat apoplexy]. AB - OBJECTIVE: To study the change of levels of plasma heat stress protein 70 (HSP70) and cytokine and their interrelationship in pathogenicity of heat apoplexy. METHODS: Plasma HSP70 was determined with Western blot, and plasma interleukin-2 (IL-2), soluble interleukin-2 receptor (sIL-2R) and interleukin-6 (IL-6) were determined with enzyme-linked immunosorbent assay (ELISA). RESULTS: Plasma level of HSP70 was 4 211.2 +/- 1,286.2 (integral optical density) in average in the patients of heat apoplexy group, 4,137.8 +/- 1, 207.5 in those of severe heat apoplexy group, and 6,043.5 +/- 1,354.8 in control group, with very significant difference (P < 0.01). Plasma IL-2 level was (64.3 +/- 32.3) pg/ml and (200.0 +/- 50.0) pg/ml and sIL-2R (54.9 +/- 33.3) U/ml and (167.5 +/- 89.0) U/ml in the patients with heat apoplexy and in controls, respectively, with very significant difference. But, there was no significant difference in IL-6 between varied groups. CONCLUSIONS: The above-mentioned results suggest that establishment of heat acclimatization correlates to the interaction of network of neuroendocrine cytokine-HSP, in which decrease in plasma IL-2 was one of the important factors causing reduction of HSP70 expression. Balance of network regulation was broken by pathological factors, finally resulting in heat apoplexy. PMID- 11860889 TI - [Detection of numerical chromosome aberrations in sperm of workers exposed to benzene series by two-color fluorescence in situ hybridization]. AB - OBJECTIVE: To study the numerical aberrations of sex chromosomes in sperm of workers exposed to benzene series. METHODS: The interphase sperm from workers of exposed and control groups were hybridized using two color fluorescence in situ hybridization with X and Y(alpha)-satellite chromosome specific DNA probes, and the aneuploidy frequencies of X and Y chromosome in X and Y sperms were detected simultaneously. RESULTS: Geometric mean concentration of benzene in workplace air was 83.95 mg/m(3), one fold higher than national maximum allowable concentration. A total of 74,517 sperm nuclei in 13 benzene-exposed workers and 89,662 sperm nuclei in 13 control workers were counted. Frequencies of disomic sperm for X, Y and XY were (0.15 +/- 0.05)%, (0.13 +/- 0.05)% and (0.20 +/- 0.11)% in workers exposed to benzene series, and (0.10 +/- 0.04)%, (0.10 +/- 0.04)% and (0.13 +/- 0.06)% in control workers. There was statistical difference in disomic frequency of X sperm and overall disomic frequency between the benzene series-exposed workers (0.15% and 0.48%, respectively) and the control workers (0.10% and 0.33%, respectively). CONCLUSION: Exposed to benzene at higher concentration may induce increase in aneuploidy frequency of sperm sex chromosome in exposed workers. PMID- 11860890 TI - [Activity of serum paraoxonase and its polymorphism in healthy Chinese of Shanghai]. AB - OBJECTIVE: To study activity of serum paraoxonase and its polymorphic distribution in healthy people of Shanghai and to set a reference value for it. METHODS: Serum activities of paraoxonase, salt-stimulated paraoxonase and arylesterase were measured in 198 subjects (105 males and 93 female, aged 21 - 61 years) in Shanghai. Three phenotypes of serum paraoxonase could be distinguished based on the ratio of salt-stimulated paraoxonase activity and paraoxonase activity and the ratio of salt-stimulated paraoxonase activity and arylesterase. Its gene frequency was calculated according to Hardy-Weinberg equation. RESULTS: Activity of serum paraoxonase was 97.6 - 472.1 unit. There was no difference in serum paraoxonase activity between male and female, nor among different ages. It was found that 27 persons in low, 71 in medium and 100 in high activity. Gene frequency of low activity (PON * A) and high activity (PON * B) was 0.369 and 0.631, respectively. CONCLUSIONS: Distribution of serum paraoxonase in healthy people of Shanghai was different from that in Caucasians. PMID- 11860891 TI - [Relationship between folate, vitamin B12, total plasma homocysteine and mutation of reductase]. AB - OBJECTIVE: To study relationship between serum and red blood cell (RBC) folate, serum vitamin B(12) (VB(12)), total plasma homocysteine (tHcy), thermolabile genotypes, and activity of methylene-tetrahydrofolate reductase (MTHFR). METHODS: Thirty-four non-pregnant and healthy mothers who gave birth to babies with neural tube defects (NTDs) and 42 mothers to normal babies previously were chosen. Plasma tHcy, RBC and serum folate, serum VB(12), activity of MTHFR and thermolabile genotypes were determined for all of them. RESULTS: There was no significant difference in plasma tHcy, RBC and serun folate between case and control groups. Serum VB(12) was significantly lower in mothers of babies with NTDs than in those with normal ones. Plasma tHcy correlated with RBC folate inversely (P < 0.05). Mean plasma tHcy for thermolabile homozygous mutation of MTHFR was significantly higher than that for either heterozygous mutation or wild homozygous mutation (P < 0.001). There was no significant difference in serum folate between groups with varied MTHFR genotypes. Plasma tHcy was slightly greater in those with lower serum folate than in those with higher one, and correlated inversely with activity of MTHFR. CONCLUSIONS: Increase in plasma tHcy correlates with homozygous mutation of thermolabile MTHFR and decrease in activity of MTHFR. Higher level of folate in the body can interfere the relationship between plasma tHcy and activity of MTHFR. PMID- 11860892 TI - [Effect of chrysotile and cigarette smoking solution on DNA damage and repair in human alveolar epithelial cells]. AB - OBJECTIVE: To study the effect of chrysotile and solution of cigarette smoke separately or combined on DNA damage and repair. METHODS: Single-cells gel electrophoresis assay was used to detect break in DNA strand and to analyze its repair function in human alveolar epithelial cell line A549, which were exposed to chrysotile or cigarette smoke solution separately or combined for one hour and then immediately cultured for different time. RESULTS: Break in DNA strands was induced obviously in A549 cells after exposure to 40 microg/ml of chrysotile or 2.5 x 10(-4) cig/ml of cigarettes smoke solution for one hour. Repair for DNA break could be seen clearly in chrysotile-treated cells after continuing culture for three hours, and 30.6% had been repaired after culture for four hours. Repair could also be seen clearly in cigarette smoke solution-treated cells after continuing culture for two hours, and 65.2% had been repaired after culture for four hours. Repair of DNA in cells exposed to both chrysotile and cigarette smoke solution combined was similar to that exposed to cigarette smoke solution only. CONCLUSION: Damage to DNA caused by chrysotile is more difficult to repair than that induced by cigarette smoking solution. PMID- 11860893 TI - [Studies on gene mutation and micronucleus formation induced by ethylnitrosourea (ENU) in transgenic mice in vivo]. AB - OBJECTIVE: To determine the feasibility of using pUC 118NX plasmid-based transgenic mice in the study of induced mutation in vivo with known mutagen, ethylnitrosourea (ENU). METHODS: pUC 118NX plasmid was recovered from the spleen genomic DNA of ENU-treated and untreated xylE, C57BL/6J transgenic mice with enzyme digestion and circularizing. The recovered pUC 118NX plasmid was electroporated into DH10B host cells, which were incubated in LB solid medium containing proper ampicillin overnight and then sprayed with catechol solution. Mutant could be detected by difference in yellow and white color of the stains and its xylE target gene could be sequenced for further identifying its mutation type. In addition, peripheral blood and bone marrow cells were isolated from xylE, C57BL/6J transgenic mice and micronucleus frequency (MNF) induced by ENU was observed. RESULTS: The spontaneous mutant frequency for xylE gene in the spleen of xylE, C57BL/6J transgenic mice was less than 4.79 x 10(-5), significantly different from that treated with ENU (50 mg/kg), 19.83 x 10(-5). Types of gene mutation induced by ENU in the spleen of xylE, C57BL/6J transgenic mice included transversion (50%), one or two-base insertion (37.5%) and transition (12.5%). MNF in peripheral blood normochromatic erythrocyte (NCE) and in bone marrow polychromatic erythrocyte (PCE) of ENU-treated (50 mg/kg x 5) mice were 7.6 per thousand and 8.8 per thousand, respectively, both significantly different from the controls treated with solvent (P < 0.01). It indicated that chromosome aberration could be induced by ENU. CONCLUSION: The pUC118NX plasmid based xylE, C57BL/6J transgenic mice, which could be used in detecting gene mutation and chromosome aberration in vivo simultaneously, provided a novel detection system for overall assessment of genetic toxicity of chemicals. PMID- 11860894 TI - [Studies on chrysotile-induced cell transformation and its intervention effect]. AB - OBJECTIVE: To explore the effects of several kinds of chemicals on chrysotile induced cell transformation in human embryo lung (HEL) cells. METHODS: After being soaked in aluminum citrate, mixed rare earth or sodium selenite solutions for one hour, chrysotile was incubated with HEL cells. Then, the number of focus of cell transformation was observed. RESULTS: Chrysotile could cause morphological transformation of HEL cells in a dose-dependent pattern, with (0.60 +/- 0.55), (1.20 +/- 1.31), (2.60 +/- 1.14) and (3.60 +/- 1.14) foci of HEL cell transformation per plate after treatment with 0.625, 1.25, 2.5 and 5.0 microg/ml of chrysotile, respectively, and there was significant difference between the groups treated with 2.5 and 5.0 microg/ml of chrysotile and control group. It indicated that chrysotile could cause morphological transformation of HEL cells. Number of focus of HEL cell transformation induced by chrysotile that was pretreated with the above-mentioned three kinds of chemicals reduced significantly, as compared with that in the group without pretreatment of it. Its inhibitory rate was 50% in the group treated with mixed rare earth, with significant difference. CONCLUSION: Pretreatment for chrysotile with these three kinds of chemicals could reduce its carcinogenicity. PMID- 11860895 TI - [Effects of chronic exposure to lead on long-term potentiation in hippocampal CA1 area of young rats in vivo]. AB - OBJECTIVE: To study the effects of chronic exposure to lead on learning and memory function in young rats, based on the plasticity of their synapse function. METHODS: With method of extracellular glass microelectrode recording, effects of exposure to lead at different concentrations on long term potentiation (LTP) induction in hippocampal CA(1) area of young rats in vivo were observed. RESULTS: Incidence of LTP and mean amplitude increase after application of high frequency stimulus decreased, as blood lead in rats reached (3.28 +/- 0.88) micromol/L, even causing long term depression. To the exposed rats, their mean increase ratio in their population spike amplitudes after high-frequency stimulation correlated inversely with their brain lead load. CONCLUSION: Chronic lead exposure could damage in vivo LTP induction and maintenance in the CA(1) area in hippocampus of rats, and the severity of damage increased with the extent of exposure to lead. PMID- 11860896 TI - [Detection of C gene mutation strain in vertical transmission of hepatitis B virus and its significance]. AB - OBJECTIVE: This study aims at detecting C gene mutation strain in vertical transmission of hepatitis B virus (HBV). METHODS: HBV C gene nt 2,022 - 2,321 was directly sequenced. RESULTS: Homology of HBV nucleotide sequence and amino acid sequence in mothers, fathers and their fetus was 100%. Mutation of 2,189 and 2,288 nucleotide of C gene resulted in substitution at the 97th and 130th amino acids. CONCLUSION: C gene mutation strain is present in vertical transmission of HBV. Presence of mutation strain may be the cause of failure in vaccination. PMID- 11860897 TI - [Effects of excessive iodine on activity of enolase in hippocampus of developing mice]. AB - OBJECTIVE: To study the effects of excessive intake of iodine for a long term on activity of enolase in hippocampal tissues of young mice. METHODS: A hyperiodine goiter animal model was replicated with drinking water with high level of iodine. Activity of neuron specific enolase (NSE) in hippocampal tissues was determined with enzyme-linked immunosorbent assay (ELISA) for young offspring of the first and the second generations of mice with hyperiodine goiter on the 7th, 14th and 21st day of age, respectively. RESULTS: Activity of enolase in young offspring of the first and second generation of mice with hyperiodine goiter was all lower than that in controls from the day of their birth to the 30th day after birth, and that at hippocampal tissues in young mice of the second generation was significantly lower than that of the first generation on the 14th day of age. CONCLUSION: Hyperiodine can decrease activity of NSE in the brain of mice. It suggests that hyperiodine can affect NSE, and thus interfere energy supply in the brain, resulting in disturbance of neuron development and brain function. PMID- 11860899 TI - [Characteristics of producing-fumonisin and dimorphic fungus of fusarium moniliforme]. AB - OBJECTIVE: To study the carcinogenesis of fusarium moniliforme and its fumonisins in animals. METHODS: The producing-fumonisin and dimorphic fungus pathogenicity of F. moniliforme from Zhejiang, China were identified and characterized. RESULTS: The F. moniliforme isolated from foodstuffs was shown to produces fumonisin B(1) > 400 microg/g in medium by IC-ELISA. Ten foodstuffs samples showed (2% - 100%) detection rate of F. moniliforme and contained fumonisin B(1) between (3.7 - 143.1) microg/g. The strains of F. moniliforme formed yeast-like colony in Sabouraud's agar plates contained 9% NaCl at 37degrees C incubator and budding multiplication mostly. In blood agar plates the strains of F. moniliforme showed grass green haemolytic reactions. CONCLUSION: It is important to carry out (or to perform) oulepidemiologicol survey of F. moniliforme as a primary pathogenic fungus, as well as the mycotoxins of F. moniliforme in vitro. PMID- 11860898 TI - [Studies on schedules for human rabies immunization]. AB - OBJECTIVES: To study the feasibility of interferon (INF) plus an immunization schedule of two dose-two dose-one dose (2 - 2 - 1) of rabies vaccine and a simple 2 - 2 - 1 schedule, and to explore immune regulation mechanism of INF. METHODS: For an IFN plus 2 - 2 - 1 schedule, 5.0, 5.0 and 2.5 I.U. of primary hamster kidney cell rabies vaccine (PHKCV) were injected on the 1st, 7th and 14th day, respectively, and 0.2 million I.U. of IFN-alpha on the 1st day simultaneously. For a simple 2 - 2 - 1 schedule, only 5.0, 5.0 and 2.5 I.U. PHKCV were injected on the 1st, 7th and 14th day, respectively. And, for controls, a routine schedule recommended by WHO was used. RESULTS: On the 7th day after immunization, their geometric mean titer (GMT) of antibody was 1.71, 1.57 and 1.21 for the above three schedule groups, respectively; on the 14th day, 62.77, 58.79 and 28.96; on the 45th day, 76.64, 72.90 and 62.22. Conversion rate of antibody was 85.00%, 61.11% and 65.00%, respectively on the 7th day, and was 100.00% for all on the 14th day. Proportion of those with protective antibody level was 10.00%, 0.00% and 0.00%, respectively for the three schedule groups, on the 7th day; 100.00%, 100.00% and 95.00%, on the 14th day; and was 100.00% for all on the 45th day. Proportion of those with antibody titer more than 1:1 was 55.00%, 33.33% and 20.00%, respectively, on the 7th day. Adverse reaction rate was 0.00% - 55.00%, without significant difference between three schedule groups. CONCLUSIONS: Lower dose of IFN could enhance the effect of immune response to rabies vaccine. IFN plus 2 - 2 - 1 rabies vaccine schedule was better than that of simple 2 - 2 - 1 schedule, while the later was better than the routine schedule. PMID- 11860900 TI - [Cytochrome P450IA1 and the genetic susceptibility to esophageal carcinoma]. AB - OBJECTIVES: To explore the susceptibility to esophageal carcinoma (EC) in different population with different cytochrome P450 IA1 (CYPIA1) genotype, and study the synergistic effects between CYPIA1 and smoking. METHODS: A case-control study (case = 111, control = 114) was used to compare the frequency of CYPIA1 genotype between EC and controls with a PCR method. RESULTS: The distribution of three genotypes of CYPIA1 in EC and controls was significantly different (chi(2) = 19.35, P < 0.01). There was a synergistic effect between the mutation of CYPIA1 4,889 site and smoking (SIA = 1.56), and a significant dose-response effect was observed. The risk of individual whose CYPIA1 4,889 site is mutated was 3 times higher than those not mutated. CONCLUSIONS: The mutation of CYPIA1 ,89 site maybe one of important host susceptible factors of EC, and the risk would increase significantly in smokers. PMID- 11860901 TI - [Effects of methylmercury on intracellular free calcium content in cerebral cortical neurons]. AB - OBJECTIVE: This study was conducted to explore effects and potential mechanism of methylmercury on concentration of intracellular free calcium (Ca(2+)) in the acutely isolated cerebral cortical neurons. METHODS: Calcium ion indicator Fura-2 two wave-length fluorophotometry was used to measure concentration of Ca(2+). RESULTS: (0.10 - 5.00) x 10(-6) mol/L of methylmercury could increase level of free intracellular Ca(2+) in cerebral cortical neurons significantly (P < 0.01), in a dose-response manner. Increase in intracellular free Ca(2+) of neurons caused by methylmercury associated with large influx of Ca(2+) outside cells and release of stored Ca(2+) inside cells, especially by influx of Ca(2+) outside cells. Influx of Ca(2+) outside cells associated with Ca(2+) channels regulated by receptor gate and potential gate, but not associated with sodium-dependent channel. CONCLUSION: Methylmercury lead to abnormal increase of intracellular free Ca(2+) in neurons, which is closely related to Ca(2+) channel. PMID- 11860902 TI - [A preliminary meta-analysis of 36 studies on impairment of intelligence development induced by iodine deficiency]. AB - OBJECTIVES: Since 1980 s', numerous studies on intelligence quotient (IQ) have documented, in the areas prevalent with severe, moderate and even mild iodine deficiency, which is a risk factor for retardation in mental development there. It was purposed for this paper to quantify the relationship between iodine deficiency and mental development in children and to explore the etiological role of iodine deficiency in mental retardation and the protective effects on children's intelligence of iodine supplementation. METHODS: A total of 59 independent investigations published during 1980 and 1998 were selected for meta analysis, including 20 studies on intelligence determined by Raven's Test and 39 by China Benit Scale. RESULTS: Homogeneity test showed that there was no significant difference in baseline features between two groups (P > 0.05). The hypothesis testing showed that the homogeneity of each study group was obviously statistically significant (P > 0.05). The results demonstrated that IQ in children at risk for iodine deficiency showed a marked drop by 8.94 points with Raven's Test and by 10.80 points with China Binet Scale, respectively, with an average drop of 10 points. Substantial evidence now available has showed that mental retardation can be prevented by effective correction of iodine deficiency through iodine supplement either iodized salt or iodized oil, which is confirmed by an obvious increase in 11.5 points of IQ in average, and in 11.85 by Raven's Test and 11.64 by China Binet Scale, respectively. CONCLUSIONS: Iodine nutrition plays an important and positive role in brain development. Iodine deficiency leads to loss of 10 points of IQ and 11.5 points can be gained for children in the iodine deficiency areas after significant iodine supplement. PMID- 11860903 TI - [Cost-effectiveness analysis of Beijing Fangshan cardiovascular prevention program in 1992 - 1997]. AB - OBJECTIVE: To determine whether a community-based cardiovascular disease (CVD) intervention program, undertaken over six years, was cost-effective. METHODS: Based on Beijing Fangshan Cardiovascular Disease Comprehensive Prevention Program, the cost for intervention and expenditure saved from caring for CVD in the communities with intervention from 1992 to 1997 were calculated, and cost effectiveness analysis was performed using disability-adjusted life years (DALYs) gained as an indicator of effectiveness. RESULTS: The cost for one DALY gained reduced gradually from 1992 to 1997, with an average ratio of cost to effectiveness of four to one (4:1). It cost annually 1,586.00, 1,380.20, 2,350.80, -905.30, -1,495.60 and -1,766.70 RMB yuan for one life-year saved, from 1992 to 1997 respectively, in a gradually decreasing trend with the increase in length of intervention. After intervention for two years, ratio of cost to effectiveness has become negative since 1994, which meant a positive benefit from intervention. Sensitivity analysis showed that ratio of cost to effectiveness was little sensitive to the changes in discount rate, weight of age and increase in cost of hospitalization for stroke and coronary heart disease, which reflected its reliability. CONCLUSION: Community-based comprehensive intervention for CVD in rural population is cost-effective. PMID- 11860904 TI - [Genetic epidemiological study on non-insulin dependent diabetes mellitus]. AB - OBJECTIVE: To study the general genetic pattern of non-insulin dependent diabetes mellitus patients. METHODS: 1,608 children were investigated for NIDDM family history, and 280 nuclear families were collected. RESULTS: The prevalence rates of NIDDM among first-degree relatives (2.38%), the parents (26.00%), the siblings (2.44%) and the offsprings (1.24%) were higher than that in general population respectively. The s/q was 10.17 by Penrose method, which was close to 1/q(1/2). The p(0) was 0.0244 by simple segregation analysis, which was lower than 0.10. The heritability of NIDDM was 0.54, using the Falconer Threshold Model. CONCLUSION: NIDDM has a familial aggregation, but not fit to the mono-genetic models. NIDDM has the feature of multifactorial inheritance. PMID- 11860905 TI - [Mutation of hepatitis B virus DNA pre-C region in patients with primary hepatocellular carcinoma in Guangxi]. AB - OBJECTIVE: To study the association between mutation of hepatitis B virus (HBV) pre-C gene and HBV infection in patients with primary hepatocellular carcinoma (PHC) but negative hepatitis B e antigen (HBeAg) in Guangxi Province, China. METHODS: Nested polymerase chain reaction (nPCR) was used for amplification of HBV DNA Pre C region in sera collected from 16 patients with PHC in Guangxi, and then their HBV DNA nPCR products were sequenced by Sanger method. RESULTS: Sera in fourteen of 16 patients showed positive HBV DNA, with a positive proportion of 87.5% (14/16). One (C23) of them was positive for HBeAg; one (C24) was negative HBeAg, but with normal sequence in his Pre C region; two (C7, C14) were co infected with HBV wild and mutant strains; and the remainder eight cases (C3, C4, C5, C8, C10, C11, C12, C13) were infected with mutant virus strains, but stop codon at codon 28 was only found in C4, C5 and C12. One case (C8) was co-infected with both HBV stop codon mutant and non-stop codon strains. CONCLUSION: It is uncommon for patients with PHC in Guangxi infected with HBV mutant strain with classical mutation at nt 1 896, which suggests that maybe there exist other types of mutation other than that in pre-C region causing HBV infection without HBeAg. PMID- 11860906 TI - [Effects of B-ultrasonic radiation during pregnancy on reproductive outcome]. AB - OBJECTIVE: To study the effects of B-ultrasonic scanning during pregnancy on reproductive outcome, including pregnant outcome and neonatal status. METHODS: Data of reproductive outcome and B-ultrasonic examination were extracted from the questionnaires and obstetric records of 12,224 primigravidas in eight hospitals of Beijing during May 1992 to April 1993. Their reproductive outcomes were analyzed according to frequency and gestational weeks they had B-ultrasonic examination taken, adjusted for potential confounding factors, such as age at pregnancy, smoking and drinking during pregnant period and history of spontaneous abortion, by logistic regression. RESULTS: There was no statistically significant difference in reproductive outcomes between groups, who had B-ultrasonic examination taken for two, three and four times or more and who had only once, respectively, who had more than four times and who had less than three times, and who had their first examination before and after the eighth week of gestation, or before and after the 12th week of gestation. CONCLUSION: No adverse reproductive outcome due to exposure to diagnostic dose of B-ultrasound radiation during pregnancy has been found. PMID- 11860907 TI - [Effect of green and yellow vegetables on serum carotenoid in children]. AB - OBJECTIVE: To assay the effect of green and yellow vegetables on serum carotenoid concentration and its relation to serum retinol level in children. METHODS: A ten week green and yellow vegetable intervention study was conducted in children aged 5.3 to 6.4 years of two classes in a kindergarten in Tai'an City of Shandong Province in early September 1996, one class of light-colored vegetable supplemented with daily 193 grams of light colored vegetables, such as cabbage, Chinese cabbage, potato, cucumber, cauliflower and turnip, and 56 grams of yellow green vegetables, such as spinach, Chinese chive, carrots and red yams; and the other one of yellow-green vegetables with daily 238 grams of yellow-green vegetables and 34 grams of light colored vegetables, both without change in regular diet. Venous blood specimens were collected before and after the experiment from each of the subjects. Serum retinol and carotenoid were determined with high performance liquid chromatography. RESULTS: Serum level of carotenoid, including all-trans-beta-carotene, cis-beta-carotene, cryptoxanthin and lutein were significantly higher after intervention in yellow-green vegetable group, as compared to their baseline levels. However, in the light-colored vegetable group, all those components, including all-trans-beta-carotene, 13-cis beta-carotene and lutein, decreased significantly after intervention, as compared with those at their baseline levels. Serum carotene level correlated significantly with that of retinol, and their coefficient of correlation was greater in the winter than in the autumn. CONCLUSION: Carotenoid nutrition status can be improved by supplementation of green and yellow vegetables. PMID- 11860908 TI - [Studies on algae and microcystin pollution in source water of Yellow River in Zhengzhou city]. AB - OBJECTIVE: To study the law of development of pollution with algae and microcystin (MC) in source water of Yellow River in Zhengzhou city. METHODS: Algae and MC were detected continuously for source water samples collected from B and S Water Works of Zhengzhou in 1998. Microcystin was determined with high sensitive ELISA. RESULTS: Monthly average density of algae was 1,439 x 10(4)/L in source water of B Water Works of Zhengzhou, with peak algae density of 4,762 x 10(4)/L in August, and that of S Water Works was 1,071 x 10(4)/L. Blue-algae of higher toxicity has become dominant species in source water from S water Works. Detection rate of microcystin was 70.83% in 24 samples collected from the two water works. CONCLUSION: According to Carlson's trophic state index (TSI), as well as other indices, such as total nitrogen, total phosphorus, chemical oxygen demands (COD) and algae density in water, source water in Zhengzhou has presented characteristics of eutrophication. PMID- 11860909 TI - [Effects of excessive iodine on apoptosis and expression of bcl-2, bax genes in the cells of brain and thyroid gland in guinea pigs]. AB - OBJECTIVE: To investigate the effects of excessive iodine intake on apoptosis and the expression of bcl-2, bax genes in the cells of brain and thyroid gland. METHODS: Guinea pigs, divided into two groups at random, were fed with distilled water containing iodine 50 microg/L (a control group with appropriate iodine) and 10,000 microg/L (a trial group with excessive iodine), respectively for six months. Apoptosis rate and expression of bcl-2, bax gene in the cells of cerebral cortex, hippocampus and thyroid gland in guinea pigs were quantitatively determined by flow cytometry and immunofluorescent techniques. RESULTS: The proportion of apoptosis in the cells of cerebral cortex, hippocampus and thyroid gland was significantly higher in the group with excessive iodine intake than that in the control one (P < 0.01). The level of expression of bcl-2 gene was notably decreased (P < 0.05), but that of bax was notably increased (P < 0.01), in the cerebral cortex, hippocampus and thyroid gland in the group with excessive iodine than in the control one. The ratio of bcl-2 to bax in the cerebral cortex, hippocampus and thyroid gland of the group with excessive iodine was decreased significantly (P < 0.01) than that of the control one. CONCLUSION: Excessive iodine intake may induce apoptosis in the cells of cerebral cortex, hippocampus and thyroid gland. The over-expression of bax gene and the down-regulation of bcl 2, which can then cause the decrease in a ratio of bcl-2 to bax, appear to play an important role in the course of apoptosis induced by excessive iodine intake. PMID- 11860910 TI - [Toxicity to neural cell development of lead and its relation to glutathione]. AB - OBJECTIVE: To investigate the toxicity of lead to embryonic neural cell development and its relation to glutathione level. METHODS: Rat midbrain micromass culture method was used to observe effects of lead on viability, differentiation and glutathione (GSH) content of embryonic neural cells. RESULTS: Lead concentrations at 3.69 micromol/L and 1.03 micromol/L could inhibit neural cell variability and differentiation by 50%, respectively, both in a significant dose-response pattern. Lead also could cause increase level of glutathione. N acetylcysteine (NAC), an antioxidant, could not only reduce adverse effect of lead on GSH, but also antagonize its toxicity to cell survival and differentiation. CONCLUSION: Lead can specifically inhibit neural cell differentiation and its neurotoxic effects on brain cell development correlated to imbalance in redox status which is mainly mediated by decrease of GSH content. PMID- 11860911 TI - [Studies on micronuclei induced by colchicine and cyclophosphamide using multicolor fluorescence in situ hybridization]. AB - OBJECTIVE: To study chromosomal composition of micronuclei (MN) induced by colchicine (COL) and cyclophosphamide (CP) in mouse bone marrow erythrocytes. METHODS: Multicolor fluorescence in situ hybridization (FISH) with centromeric and telomeric DNA probes was applied to analyze chromosomal composition of micronuclei induced by COL and CP in mouse bone marrow erythrocytes. RESULTS: About 83.5% of COL-induced MN revealed both centromeric and telomeric signals. Of the CP-induced MN, 74.5% showed telomeric signals only. CONCLUSION: Majority of COL-induced MN contain whole chromosome and that of CP-induced MN mainly contain acentric fragments. Multicolor FISH with centromeric and telomeric DNA probes was a precise technique for analyzing chromosomal composition of MN. PMID- 11860912 TI - [Lung injury caused by passive smoking and its effects on cytokines in rats]. AB - OBJECTIVE: A rat model with chronic bronchitis was replicated by passive inhalation of cigarette smoking fume to study its long-term effects on lung injury and nitric oxide (NO), interluekin-6 (IL-6), interleukin-8 (IL-8). METHODS: Levels of nitrogen dioxide (NO(2)) and nitrogen trioxide (NO(3)) were measured with spectrophotometry in rats indicating their level of nitric oxide (NO). Levels of IL-6 and IL-8 were determined by enzyme-linked immunosorbent assay (ELISA). RESULTS: Levels of NO in serum, bronchial alveolar lavage fluid (BALF) and lung tissue in the smoking group were significantly lower than those in the normal controls (P < 0.01). But, levels of IL-6 and IL-8 were higher in the smoking group than those in the controls. CONCLUSION: Long-term passive smoking could cause injury of lung tissue to certain extent, reduction in secretion of NO in endothelial cells and damage to pulmonary vessels. PMID- 11860913 TI - [Toxic effects of aluminum on human embryonic cerebral neurocytes in vitro studies]. AB - OBJECTIVE: To study the toxic effect of aluminum on human embryonic cerebral neurocytes in vitro studies. METHODS: Human embryonic cerebral neurocytes were divided into three groups (control, low Al and high Al groups). The growth, development and morphology of neurocytes were observed by light and electron microscopy. The protein, DNA, lipid peroxide (LPO) contents and the activity of neuron-specific enolase (NSE) and cholinesterase (CHE) of neurocytes were detected. RESULTS: The number of viable cells in high Al group was significantly lower than that in the control group (P < 0.05). The number of swelling, degenerated and necrotic cells was significantly increased in high Al group. The DNA content and the activity of NSE and CHE of neurocytes were significantly decreased with the increase of Al concentrations (P < 0.05). The LPO content of neurocytes in the high Al group was significantly higher than that in the control group (P < 0.05). There was a positive correlation between the Al dose and LPO content of neurocytes (P < 0.05). The electron microscopy study showed that the membrane structure of cell was damaged in the high Al group and the arrangement of microtubules in the processes was disordered. CONCLUSIONS: The growth, development and function of human embryonic cerebral neurocytes was inhibited in the high Al group. The neurotoxicity of Al may be caused by lipid peroxidation and the damage of cell membrane. PMID- 11860914 TI - [Effects of oral calcium supplementation on blood pressure in population]. AB - OBJECTIVE: To assess the effects of oral intake of calcium lactate tablet on blood pressure and serum levels of sodium, potassium, calcium and phosphors. METHODS: In a rural area with natural population with higher average blood pressure and prevalence of hypertension, and with high salt and low calcium intake in diet in Yu County, Shanxi Province, 112 volunteers aged 30 to 64 years were selected based on a mass screening for hypertension, and randomized into trial and control groups with double blinding, taking calcium lactate tablets (800 mg of calcium daily) and placebo respectively for five weeks. Ninety-eight of them complied with the medical order completely, 39 males and 59 females, and were followed-up for systolic (SBP) and diastolic blood pressure (DBP), and their fasting venous blood specimen and 8-hour nocturnal urine specimens were collected. Twenty-four-hour diet recall was obtained for three consecutive days from all of them. RESULTS: Reduction of 4.7 mm Hg (0.62 kPa) and 2.7 mm Hg (0.36 kPa) in systolic and diastolic blood pressure, respectively in the trial group was observed, as compared with that in control one, with P-values of 0.027 and 0.074, respectively, after 35 days of treatment. The higher their baseline BP level, the greater reduction of their BP after treatment. Urinary excretion of calcium increased and serum level of phosphorus reduced after oral administration of calcium. CONCLUSION: In an area with higher prevalence of hypertension and with high salt and low calcium intake in diet, change in diet composition or supplementation with calcium could reduce their average blood pressure and prevent them from hypertension for those with mild and moderate hypertension and in high risk with higher blood pressure. PMID- 11860915 TI - [Efficacy and immune memory of plasma-derived hepatitis B vaccine 11 years after primary immunization]. AB - OBJECTIVE: To evaluate the long-term efficacy of hepatitis B vaccine 10 years after primary immunization to provide scientific basis for the time of revaccination. METHODS: The study was strictly designed with randomization, double-blinding, and placebo-controlled method to observe the efficacy and immune memory 11 years following hepatitis B vaccination. RESULTS: Immunogenicity and protective rate of vaccine were still kept well 11 years after immunization with a protective rate against HBV infection of 73.5%. But, there was no significant difference in HBV infection rates between vaccine group and placebo-controlled group (7.89% vs. 13.25%, P > 0.1) nine to 11 years following immunization. There still existed immune memory 11 years after immunization, but it was significantly weaker than that within the first 10 years after immunization. CONCLUSION: The efficacy of the vaccine had begun to drop 11 years after immunization, which should be followed up further to reach a clear conclusion. PMID- 11860916 TI - [Study on the protective effect of grape procyanidins on DNA damage induced by irradiation]. AB - OBJECTIVE: To study the protective effect of grape procyanidins (GPC) on the irradiated DNA damage. METHOD: In vivo study: After orally administration of GPC, mice in different groups were exposed to whole-body (60)Co gamma-rays at a dose of 2.0 Gy and the fluorospectrometry method was used to rapidly detect the DNA strand breaks of mouse spleen cells. In vitro study: Human venous blood with different concentrations of GPC was exposed to (60)Co gamma-rays at a dose of 1.0 Gy, then WBC DNA content was measured after 1, 30 and 60 min exposure, respectively. RESULTS: In the three treated groups (500, 250 and 50 mg/kg of procyanidins), DNA content of mouse spleen cells was 18.76%, 15.74% and 5.06% more than that of the positive controls respectively. DNA content of human WBC was higher than the positive controls by 17.6%, 11.1% and 3.9% increase respectively one minute after irradiation and 21.3%, 13.8% and 5.3% increase respectively 60 min after irradiation. These differences were all statistically significant. CONCLUSION: GPC has a protective effect on DNA damage caused by irradiation, and is helpful to DNA repair. PMID- 11860917 TI - [The effect of fluoride-arsenic exposure on the lipid peroxidation and antioxidation of the offspring of rats]. AB - OBJECTIVE: To provide information on the effects of the offspring of rats exposed to fluoride-arsenic. METHODS: The levels of lipid peroxidation and the abilities of antioxidation were determined in the blood of the rats and their offsprings under two generations-one nest reproductive test. RESULTS: The activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in the blood decreased with the increase of exposure dose. For example, the activity of SOD, was 14.56, 13.74, 11.89 and 11.21 micromol x min(-1) x mg Hb(-1) in different dose groups of F(2), respectively. In contrast, the concentration of lipid peroxides (LPO) increased. Eight weeks after exposure, the activities of SOD and GSH-Px increased, the activity of SOD was 13.97, 13.55, 13.47 and 12.76 micromol x min(-1) x mg Hb(-1), respectively, and the concentration of LPO returned to normal level. CONCLUSION: Fluoride-arsenic exposure can cause oxidative damage of the rat's offspring. PMID- 11860918 TI - [Effects of phoxim and fenvalerate on TTX-S and TTX-R sodium channels in the DRG neurons of adult rat]. AB - OBJECTIVE: To study the joint neurotoxic effects of phoxim (Pho) and fenvalerate (Fen) on tetrodotoxin-sensitive (TTX-S) and tetrodotoxin-resistant (TTX-R) Na(+) currents in dorsal root ganglion (DRG) neurons of adult rat. METHODS: Whole cell patch clamp technique was used to test the effects of Pho and Fen on TTX-S and TTX-R sodium currents in DRG neurons. RESULTS: The inactivation of TTX-R sodium channel was obviously slowed down by Fen. The tau(Na) of peak currents at doses of 10, 50 and 100 micromol/L Fen and control groups were (8.10 +/- 2.41) ms, (11.78 +/- 2.76) ms, P < 0.01, (8.76 +/-1.94) ms, P < 0.05 and (6.41 +/- 1.32) ms respectively. The inactivation of TTX-R sodium channel tail currents was also significantly delayed by Fen. The tau(Na) of the tail currents at doses of 10, 50, 100 micromol/L Fen and control groups were 6.11 +/- 0.52 (P < 0.05), 7.82 +/- 0.82 (P < 0.05), 7.23 +/- 1.09 (P < 0.05) and (4.91 +/- 0.97) ms separately. As compared with TTX-R sodium channel, the TTX-S sodium channel was less responsive to Fen exposure, which only led to slowly decay TTX-S sodium tail currents. There was no any effect of Pho on the TTX-S and TTX-R sodium channels. The mixed treatment of a Pho and Fen did not show joint effect on the sodium currents. CONCLUSION: Both the peak and tail currents are changed by Fen, however, Fen has more remarkable effects on TTX-R than on TTX-S sodium channel. The combined exposure to Pho and Fen shows no joint effect on the sodium channel. PMID- 11860919 TI - [Study on beta-adrenoceptors mechanism of cadmium immunotoxicity]. AB - OBJECTIVE: To study if the beta-adrenoceptors mechanism plays important role in the immunotoxicity of cadmium. METHODS: The beta-adrenoceptor density, cellular cyclic adenosine 3',5' monophosphate (cAMP) levels, and DNA damage of rat spleen lymphocytes were studied. Beta-antagonist propranolol was used to block the beta adrenoceptor. RESULTS: In the single Cd administration group: beta-adrenoceptor density of spleen lymphocytes in the middle and high Cd concentration groups was higher than that in the non-Cd treated control group (P < 0.05), the levels of cAMP increased with the increase of Cd doses, and the result of the high Cd group was higher than that of the control group (P < 0.05). The proliferation function of T lymphocytes was lower in each Cd group than that in the control group (P < 0.05). Single cell gel electrophoresis (SCGE) detected that DNA damage of spleen lymphocytes in each Cd group were more severe than the control (P < 0.05). Pretreated with Pro group: After blocked with propranolol, beta-adrenoceptor density increased in the Pro control group and the density in all Cd treated groups was higher than the control group (P < 0.05), but all Cd treated groups showed no statistical significance than the Pro control group (P > 0.05). The cAMP levels, MTT and the subsets of T lymphocytes showed no statistical significance among all the groups (P > 0.05). The results of SCGE showed that DNA damage was most severe only in the high Cd group than other groups (P < 0.05). CONCLUSION: The results suggest that the nerve-internal secretion system plays an important role in the immunotoxicity of cadmium, and the beta-adrenoceptor mechanism is important in the immunotoxic mechanism of cadmium. PMID- 11860920 TI - [Protective efficacy of H2 strain live attenuated hepatitis A vaccines in an outbreak of hepatitis A]. AB - OBJECTIVE: To investigate the protective efficacy of H(2) strain live attenuated hepatitis A vaccine (H(2)-strain vaccine) locally produced in an outbreak of hepatitis A (HA). METHODS: A nonrandomized controlled design was used in a trial for HA vaccination. Five thousand five hundred and fifty-one children at preschool classes and at grades 1 - 3 of primary schools were selected in 14 villages. Epidemiological and serological methods were used to evaluate its protective efficacy of the vaccines and serum immunoglobulin M against hepatitis A virus (IgM anti-HAV) was detected with ELISA for 271 subjects in vaccinated group and 341 in the control one. RESULTS: An outbreak of HA occurred one year after the vaccination with 26 cases in the vaccinated group, including one vaccine-related case with an attack rate of 0.28% (1/356), and 25 cases in the control group with an attack rate of 5.92% (25/422). The protective efficacy of vaccines was 95.27% (95% confidence interval of 85.83% to 104.72%). Positivity of IgM anti-HAV, i.e., newly-infected cases, was 4.43% (12/271) in the vaccinated group and 13.38% (42/314) in the control group, respectively. Clinically apparent cases accounted for 8.33% (1/12) and 50.00% (21/42) in newly-infected cases in vaccinated and control groups, respectively. CONCLUSION: Vaccination with H(2) strain vaccines locally produced was effective in reducing occurrence of HA during its outbreak, but not so effective in preventing its infection. A booster dose of HA vaccine should be vaccinated for children in adequate time. PMID- 11860921 TI - [A cost-benefit evaluation of neonatal screening for phenylketonuria and congenital hypothyroidism]. AB - OBJECTIVE: In order to make the best use of health care resources, to achieve the maximal social and economic benefits and to lay a foundation for popularizing neonatal screening for phenylketonuria (PKU) and congenital hypothyroidism (CH), a cost-benefit analysis of the screening program was conducted. METHODS: Cost for and benefit gained from screening were calculated according to the average incidence of two diseases recommended by the Ministry of Health, mean charge for neonatal screening, sampling extraction of medical history and data published in national economic statistics. RESULTS: The cost of neonatal screening, treatment for PKU with low phenylalanine milk powder and follow up, the total add up to 128 793 Yuan However the direct and indirect financial benefits is 481 263 Yuan, ratio of cost to benefit was 1:3.7. The cost of neonatal screening for CH is 129 175 Yuan. However the financial benefits including the money saved in treatment, nursing care, special education and the loss of income avoided is 468 470 Yuan, ratio of cost to benefit was 1:3.6. CONCLUSION: Neonatal screening for PKU and CH in this country reflects a better economic and social benefit and merits further popularization. PMID- 11860922 TI - [Studies on human streptococcal infectious syndrome caused by infected pigs]. AB - OBJECTIVES: To demonstrate the clinical manifestations of S. suis infectious syndrome; to study the characteristics of causative organism and its source. METHODS: 25 cases of unknown causes of food poisoning were identified in the central area of Jiangsu province, China in July 1998. Biological specimens was collected in some patients, and the causative organism was isolated using a number of different culture medium. Isolated organisms were identified by serum antibody tests, API biochemical tests, drug sensitivity tests and animal model experiments. The human born strains and the pig born strains were compared by RAPD technique. Epidemiological methods were applied to trace the source of causative organisms. RESULTS: Cases were clinically categorized into two types: Streptococcal toxic shock syndrome (STSS) and streptococcal meningitis syndrome (SMS). Three strains of streptococci were isolated respectively from the blood and cerebrospinal fluid of 6 cases. All these 6 strains were confirmed to be S. suis type 2 by culture characteristics, morphology, biochemical characteristics, serum antibody tests and fatty acid profile analysis. Animal model experiment showed that these strains were sensitive to rabbits and pigs, but not to rats. RAPD fingerprint test revealed an identity between human born and pig born strains. CONCLUSIONS: The aetiological agent of these human cases was S. suis type 2. Human cases were contracted by direct contact with the infected pigs or the corpse of infected pigs. PMID- 11860923 TI - [Genotyping of hepatitis C virus and homology analysis of its core gene sequences isolated from different geological regions]. AB - OBJECTIVE: To study the structure of its genotypes and gene variation of hepatitis C virus (HCV) isolated from different geological regions. METHODS: Serum specimens were collected from patients with HCV infection of Henan and Hebei Provinces of China, as well as of Sweden and Germany. HCV RNA in their serum was detected by retro-transcription polymerase chain reaction (RT-PCR) and genotyped. The core gene of 11 HCV isolates were sequenced and their homology was analyzed for different geographic regions with software. RESULTS: Structure of HCV genotypes, isolated from two provinces of China, and from Sweden and Germany, was not identical. The homology of their core gene sequences of the same HCV genotype isolated from the same region was higher than that from different regions, and subtypes 1b and 2a were found in serum collected from China and subtypes 1a, 2c and 3a in serum from Sweden and Germany. CONCLUSION: Structure of HCV genotypes associated with their geographic distribution and difference in sequences of the same HCV genotype also associated with their geographic regions. PMID- 11860925 TI - [Relationship between blood lipid level and apolipoprotein E gene polymorphism in offspring from longevity and non-longevity families]. AB - OBJECTIVE: To study relationship between genetic factor and level of blood lipid. METHODS: Levels of blood lipid and difference in apolipoprotein E (apo-E) allele frequencies were compared in 86 offspring in seven Uigur longevity families and 62 offspring in nine Uigur non-longevity families and their relationship was explored. RESULTS: After adjustment for age and occupation, blood indicators related to atherosclerosis were all lower in the offspring of longevity families than in non-longevity families, and those correlated inversely with it were higher in the former. Blood levels of triglyceride and apo-B were significant lower (P < 0.05) and the ratio of HDL-ch to LDL-ch was significantly higher (P < 0.05) in male offspring of longevity families than of non-longevity families. The ratio of apo-A to apo-B was significantly higher in female offspring of longevity families than of non-longevity families. Frequency of epsilon4 allele also was significantly fewer in the offspring of longevity families than of non-longevity ones (P < 0.01). Also, analysis showed that individuals with epsilon4 allele had higher levels of serum total cholesterol, lower density lipoprotein cholesterol and apo-B than those with other alleles. CONCLUSION: Average level of blood lipid was better in the offspring of longevity families than that of non-longevity families. apo-E allele polymorphism can in part explain its difference. PMID- 11860924 TI - [Investigation on E. coli O157 in Fujian, China]. AB - OBJECTIVES: To understand the distribution of Escherichia coli O157 (E. coli O157) in human beings, livestock and fowls in Fujian Province, and biological features of the isolated strains. METHODS: Fecal specimens from diarrheal patients, livestock and fowls and meat specimens in Putian and Fuzhou were collected for detection of E. coli O157 during 1997 - 1998. RESULTS: A total of 2,725 specimens were detected, and 76 strains of E. coli O157 were isolated from pigs, pigeons, cattle, chickens, ducks, et al., as well as from the diarrheal patients. These strains could be classified into three types according to their serological tests, including 33 strains of O157: H7, 21 strains of O157: NM, and 22 strains of O157: H?. Detecting rate of E. coli O157 from pigs was the highest. The isolated strains could cause diarrhea in human and obvious pathological changes in the intestines of mice, but could not produce cytotoxin Vero, or heat stable or heat-liable enterotoxin LT or ST. CONCLUSIONS: There were three types of E. coli O157 in Fujian. But, pathogenic factors of O157: H7 and O157: NM were different from those reported abroad. It suggested that there possibly existed some new pathogenic factors. PMID- 11860927 TI - [Selection of population-based controls in pair-matched case-control study analyzing risk factors for lung cancer]. AB - OBJECTIVE: To explore the methods to select population-based controls in a pair matched case-control study of risk factors for lung cancer and to lay foundation for development of their monitoring in urban areas. METHODS: Data of cases of lung cancer and their three different groups of pair-matched control based on population from the "Study on surveillance for incidence and mortality of common malignant tumor and their risk factors" in Beijing, Tianjin, Shanghai and Chongqing were personally-interviewed and analyzed with conditional multivariate logistic regression using STATA software involving 1 312 pairs aged 35 - 74. RESULTS: Odds ratios for main risk factors were consistent with the three groups of controls, and there was certain difference in ORs for dietary factor, etc. between the studies with three difference control groups. There was little differences in frequencies of exposure to risk factors between studies with neighbor-controls and normal-controls. Cigarette smoking, lower body mass index, long-time depressed mood, lower capability of emotion regulation, previous respiratory diseases, family history of cancer in the first-degree relatives significantly increased the risk for lung cancer after adjusted for age, education level, family average income and other risk factors. CONCLUSION: It was feasible to use population-based controls instead of neighbor controls in surveillance for risk factors of lung cancer. PMID- 11860926 TI - [Effect of carbon disulfide on pyrogallol- luminol auto-oxidation chemiluminescence system and superoxide anion]. AB - OBJECTIVE: To study the effect of carbon disulfide (CS(2)) on pyrogallol-luminol auto-oxidation chemiluminescence (PA-L) system and production of superoxide anion. METHODS: Luminescence dynamic curve of CS(2) and inhibition effect of superoxide dismutase (SOD) on intensity of chemiluminescence induced by CS(2) were studied with PA-L chemiluminescence system. RESULTS: Different concentrations of CS(2) (10, 40, 80 mg/ml) could enhance the emission intensity of chemiluminescence of PA-L system and delayed its peak time, with a significant dose-effect relationship, as compared with that of solvent ethanol. Peak of chemiluminescence induced by 80 mg/ml of CS(2) was higher than its background value without solvent ethanol. Chemiluminescence induced by CS(2) could be inhibited by SOD. CONCLUSION: CS(2) can generate superoxide anion in PA-L system and delay the peak time of chemiluminescence emission dynamic curve. PMID- 11860928 TI - [Trend and causes of injury deaths among children in Macheng city of Hubei province during 1969 through 1998]. AB - OBJECTIVES: To describe time trend of mortality and proportional mortality of injury and to determine the characteristics of distribution of injury deaths in children aged 0 - 14 in Macheng city during 1969 to 1998. METHODS: Data were collected from Notification System for Deaths of Macheng City during 1969 to 1998. Injury claimed 14,510 deaths caused by injury in children in total during these years. Changes in mortality and proportional mortality for injury were fitted with log-linear and simple linear regression models to compare their difference in children with different gender and ages. RESULTS: Both injury mortality and proportional mortality in infants declined during the past 30 years, and injury mortality dropped but proportional mortality due to injury increased gradually in children aged 1 - 14 years old. From 1984 to 1998, the average injury mortality was 81.36 per 100,000 children of 0 - 14 of age, with a potential years of life lost rate was 57.92 per 1,000 children. Boys had higher injury mortality than girls. Injury mortality in infants reached 560.15 per 100,000, and mechanical suffocation was the leading cause of injury deaths in infants. Injury mortality was 95.48 per 100,000 in children aged 1 - 4 years. Injury deaths in children aged 0 - 4 accounted for 73.2 percent of total injury deaths. And, drowning was the leading cause of injury deaths in children aged 1 - 14 years. CONCLUSION: Children under five years old were the vulnerable population for injury death needed for special care and strategy and measures should be taken to control injury in children in the rural areas. PMID- 11860929 TI - [A study on incidence of injury and its socio-economic loss in children and young adults]. AB - OBJECTIVES: To study the current status of incidence of injury among children and young adults and the causes of common injuries, and to estimate its socio economic loss and extent of harmfulness. METHODS: Pupils in 19 primary and middle schools aged 7 to 18 years, totaling 14,533, were recruited with stratified cluster sampling during 1998 to 1999 in Guangzhou, Maoming, Jiangmen and Shantou. Judgement for injury was based on the following criteria: (1) diagnosed and treated in hospitals or school clinics, (2) a half-day off or more due to injury, and (3) emergency management by pupils' parents or teachers. RESULTS: There were 6 941 pupils suffered from varied injures during the year, with an incidence rate of 47.76% (50.08% for boys and 45.02% for girls). Incidence rate of injury was higher in the middle school pupils (13 - 18 years old, 58.49%) than that in the primary school pupils (7 - 12 years old, 40.08%). The incidence increased significantly with age, with a peak at ages of 13 - 15. Major causes of injuries resulted from falls, injury by sharp articles, collision, traffic injuries and burn/scalds, etc, which usually occurred due to carelessness in sports, playing, walking, bike-riding and working. Frequency of multiple injuries related to the educational level of parents and depended on whether or not an only-child in family. There were 2,173 injured pupils (accounting for 31.3% of the total) visited clinics or emergency department in hospitals and 627 (9.0%) hospitalized for treatment. Twenty-eight percent of the injured pupils were absent from school, with an average absenteeism of 5.6 days. There were 154 injured pupils with transient disability and 53 with permanent disability, with a disability rate of 410.47 per 100,000. Cost for their medical care averaged 81.5 yuan RMB per injured pupil. CONCLUSIONS: Injury was a common and frequently-happened incident among children and young adults and could seriously affect their health, development and growth, studying and their future of children and young adults. Health education, safety-promotion and removing hidden danger for injury are the major measures to prevent from injury, and the government and the whole society should pay great attention to injury issues in children and young adults. PMID- 11860931 TI - [A comprehensive evaluation on intervention measures for injuries in primary and middle school students]. AB - OBJECTIVE: To explore the intervention measures for prevention and control of injuries and evaluation for their effectiveness in students of middle and primary schools. METHODS: Effectiveness of intervention was evaluated by comparison of incidence of injury before and after intervention in 3,896 students of nine middle and primary schools and comparison with a non-intervention group in 651 students of four primary schools in Jiangmen City, Guangdong Province during January to December, 1999. RESULTS: Incidence of injury dropped to 11.78% after intervention from 50.55% before intervention in the intervention group, with 76.70% of reduction. Proportion of severe injury dropped to 0.65% after intervention from 7.84% before intervention. Incidence of multiple injuries dropped to 3.15% from 19.16%, with 83.56% of reduction. But, in the control group, incidence of injury dropped to 49.00% from 52.67%, with 6.97% of reduction only, with a ratio of reduction of intervention to control of 11.02% and a ratio of cost to benefit of 1:13.9. CONCLUSION: Intervention measures focusing on health promotion can prevent and control the occurrence of injury in students of middle and primary schools cost-effectively. PMID- 11860930 TI - [A cross-sectional study on injuries in the residents of Shijiazhuang city]. AB - OBJECTIVE: To comprehend current status of injury in the residents of Shijiazhuang city and to lay a foundation for prevention from injury. METHODS: A retrospective study on injury in the 21,544 residents of two and six rural villages was conducted by multi-stage stratified sampling. RESULTS: Standardized incidence rate of injury was 4.21% in Shijiazhuang city, higher in the rural than in the urban areas and higher in men than in women. The top three causes of injuries were traffic accidents, falls and poisoning. Traffic accidents and poisoning ranked the first place in the urban areas and in the rural areas, respectively. Falls occurred mostly in those aged less than 14 years and more than 60 years. Gas poisoning and traffic accidents occurred more often in those aged 15 - 34 years and traffic accidents ranked the first in the age group of 35 59 years old. Standardized mortality rate of injury was 48.78 per 100,000, higher in the rural than in the urban areas. Standardized mortality rate of suicide was the highest with 23.21 per 100,000 in the countryside and that of traffic accidents ranked the next. In average, 22 years of potential life lost, 17 work years of potential life lost and nine valued years of potential life lost for each death were caused by injury. Disability rate caused by injury was 1.1 percent. Direct economic loss for each case with injury cost 679.98 RMB yuan. Incidence rate for traffic accidents ranked the highest and the most severe in all injuries. CONCLUSION: Injury has become an important public health problem harmful to the people's health. PMID- 11860932 TI - [Study on the measurement of skinfold thickness and estimation of body composition in Chinese primary and secondary school students]. AB - OBJECTIVE: To determine the patterns and characteristics of skinfold thickness and body composition of Chinese primary and secondary school students. METHODS: 82,921 Han youths aged 7 through 18 were randomly selected from 13 provinces. Their biceps and subscapular skinfolds were measured and the body compositions were estimated by using Changling's and Brozek's models. RESULTS: In boys, the biceps skinfolds reached the top of 10.9 mm, and then gradually decreased. The subscapular skinfolds increased rapidly from 5.7 mm in aged 7 to 8.7 mm in aged 12, and then slowly increased to 10.8 mm in aged 18. The biceps and subscapular skinfolds of girls gradually increased from 8.9 mm and 6.0 mm in aged 7, to 17.0 mm and 16.5 mm in aged 18, respectively. Their increments were especially large during the post-puberty period. The percentage of fat in boys increased from 14.0% in aged 7 to the relatively higher levels (16.1% - 17.6%) during aged 10 - 14, and then decreased significantly to about 13% in the 15 - 18 age groups. The percentage of fat in girls significantly increased from 19.6% in aged 7 to 23.0% in aged 12, and reached 25.6% in aged 18. The percentage of fat of the girls was significantly higher than that of the boys in all age groups. The lean body mass, both of the boys and girls, gradually increased from age 7 to age 18, but the absolute increments of boys were 12.1% higher in aged 7, 16.8% higher in aged 14, and 34% higher in aged 18 than that of girls, Significant differences of skinfold thickness were found not only among different areas and socioeconomic groups, but also between urban and rural populations. The percentile norms of the sum of skinfold thickness were set up, both for boys and girls, respectively. It was found that the P(50) of the norms were evidently lower than that of the Japanese, which showed the generally low level of Chinese students' skinfolds. But the differences of skinfolds between Chinese and Japanese in the high percentiles, such as the P(90), P(95) and P(97), were small, which suggested the subcutaneous body fat of many Chinese adolescents had already reached a quite high level. CONCLUSIONS: It will be a big challenge to prevent obesity among Chinese students in the near future. The percentile norms set up in this study cover various age and sex groups of Chinese students. These norms will be useful for further screening obesity in Chinese students. PMID- 11860933 TI - [Direct effects of fluoride on activities of bone morphogenetic protein]. AB - OBJECTIVE: To demonstrate the direct effects of fluoride on activities of bone morphogenetic protein (BMP). METHOD: A complex sample containing BMP and sodium fluoride was implanted under the abdominal skin of young male Wistar rats. The histological characteristics, and activities of alkaline phosphatase and acid phosphatase of the implanted were examined to study the effects of fluoride on biological activities of BMP. RESULTS: More fibroblasts cell were transformed to osteoblasts cell in animals treated by BMP with fluoride than those treated by BMP only. During bone formation induced by BMP, fluoride increased the activities of osteoblasts and accelerate mineralization, while inhibited the activities of osteoclast. CONCLUSION: Fluoride can affect biological activities of BMP directly. PMID- 11860936 TI - [Drinking water types, microcystins and colorectal cancer]. AB - OBJECTIVE: To assess the relationship between microcystins (a blue-green algal toxin) in drinking water and colorectal cancer. METHODS: Eight townships were randomly selected as study sites in Haining city of Zhejiang province, China. Four hundred and eight colon and rectum carcinoma cases diagnosed from 1977 to 1996 were identified from cancer registry in the study sites. A retrospective survey on types of drinking water of all 408 cases was conducted. Population data and drinking water sources data were provided by local household registration and local health institution, respectively. Water samples from different sources (well, tap-water, river and pond) were collected and microcystins concentrations were measured by an indirect competitive ELISA method. RESULTS: The incidence rates of colorectal cancer were significantly higher among people drinking river or pond water than those who drank well water or tap-water in both males and females. And comparing with other sources to well water, the relative risks (RR) were much higher for people using river water (7.94) and pond water (7.70). The positive detection rates (> 50 pg/ml) of microcystin in well, tap-water, river and pond water were 0.00%, 0.00%, 36.23% and 17.14% respectively. The highest concentrations of microcystins were 1 083.43 pg/ml (river) and 1 937.94 pg/ml (pond) in the positive samples. Microcystins concentrations in river and pond were significantly higher than the concentrations in well and tap water (P < 0.01). The Spearman rank correlation analysis showed that in the study sites, the microcystins concentrations of river and pond water were positively correlated with the incidence of colorectal cancer (r = 0.881, P < 0.01). CONCLUSION: Drinking surface water (river or pond) is one of the risk factors for colorectal cancer. Microcystins may be associated with incidence of colorectal cancer. It is suggested that further study should be carried out to clarify the relationship between colorectal cancer and microcystin in drinking water. PMID- 11860935 TI - [Effects of methyl tertiary butyl ether on cell cycle and cell apoptosis]. AB - OBJECTIVE: To explore the effects of the new gasoline additive, methyl tertiary butyl ether (MTBE) on cell cycle and cell apoptosis. METHODS: Flow cytometry was used to evaluate the effect of MTBE (1, 2, 4 microl/ml, 24 h) on NIH/3T3 cell cycles; and the effect of MTBE on Hela cell apoptosis was evaluated by detecting cell survival using crystal violet staining. RESULTS: Flow cytometry showed that MTBE could change NIH/3T3 cell cycles, decrease the number of cells in S stage, and arrest cells at G(2) + M stage. The results suggested that MTBE could affect NIH/3T3 cell cycles and induce cell proliferation. This situation existed 48 hours after the treatment, and cell cycles came back normal 96 hours after the treatment. By detecting cell survival using crystal violet staining, we found that MTBE could inhibit the apoptosis of Hela cells which was induced by tumor necrosis factor (TNF)alpha and cycloheximide. CONCLUSION: MTBE's carcinogenicity to animals may relate to induction of cell proliferation and inhibition of cell apoptosis. PMID- 11860937 TI - [A case-control study of the risk factors of lung cancer in Beijing, Tianjin, Shanghai, Chongqing metropolitan areas]. AB - OBJECTIVE: To analysis the risk factors of lung cancer and determine the surveillance index in urban areas in four metropolises in China. METHODS: The paper used the data from the in-person interviews of National Research on Malignant Tumor Morbidity, Mortality and Surveillance Methods in 1996. We carried out a 1:1 matched case-control study based on population involving 1 312 cases and 1 312 neighborhood controls aged 35 - 74 from Beijing, Tianjin, Shanghai, Chongqing urban areas. The relationships between risk factors and lung cancer were analyzed by conditional univariant and multivariate logistic regression on the computer package, STATA. RESULTS: Cigarette smoking, lower body mass index, longtime depressed mood, lower capability of emotion regulation, previous respiratory diseases, family history of cancer in first-degree relatives significantly increased the risk of lung cancer after adjusted by age, education, family average income and other risk factors. The odds ratios of these risk factors were 3.46 (95% CI 2.70 - 4.45), 1.18 (95% CI 1.22 - 1.40), 2.64 (95% CI 1.95 - 3.57), 2.71 (95% CI 2.02 - 3.65), 2.28 (95% CI 1.81 - 2.88), 1.79 (95% CI 1.38 - 2.32), respectively. The odds ratio associated with the highest quartile of fresh vegetables consumption compared with the lowest was 0.75 (95% CI 0.65 - 0.87), showed an inverse relationship. Exercising more often had a significant inverse association between cases and controls, and its odds ratio was 0.81 (95% CI 0.74 - 0.89). CONCLUSION: Cigarette smoking, psychological factor, previous respiratory diseases and family history of cancer were the main risk factors of lung cancer in four cities. They were the important surveillance index of lung cancer. The relation between lung cancer and environmental tobacco smoke, dietary factors and the factors related with female etc. should conduct further studies or special studies which defined the exposures carefully and exactly. PMID- 11860934 TI - [A study on methods to measure the social burden of disease and their significance]. AB - OBJECTIVE: To explore the effects of social burden of disease (BOD) on the total BOD. METHODS: With stroke as an example, BOD was measured using the index, disability adjusted life years (DALY), and partial social BOD was measured using another index, time lost for nursing care of hospitalized patients. RESULTS: The burden of stroke was 11.70 DALYs per thousand in men and 9.38 in women, respectively, and the burden for caring of patients with stroke during hospitalization accounted for 2.2% and 2.8% of the total BOD for men and women, respectively. CONCLUSION: Effects of social BOD should be taken into consideration in measurement of total BOD. PMID- 11860938 TI - [Risk factors of type 2 diabetes in Shenzhen]. AB - OBJECTIVE: The study was designed to investigate the risk factors of Type 2 diabetes in the new city Shenzhen. METHODS: A representative sample of 8,182 adults aged over 20 to 75 years old were randomly selected in the residents of Shenzhen, China in 1997. Type 2 diabetes was diagnosed according to the WHO criteria. Questionnaire survey, anthropometric measurement and blood analysis were carried out, and logistic regression analysis was used to study the risk factors. RESULTS: The results showed that the prevalence of type 2 diabetes in subjects with high total cholesterol were 4.9 times of those with lower TC value. Subjects with high triglyceride had a prevalence 5.7 times of those who had lower TG. Those HDL-ch had a prevalence 68.7% higher than those with normal HDL-ch value; on the other hand, those HDL-ch > 1.91 mmol/L were 44.6% lower in the prevalence than those with normal HDL-ch. The prevalence of type 2 diabetes in subjects with BMI >or= 25 were 2.6 times of those with BMI < 25. Subject with hypertension (>or= 140/90 mm Hg) had a prevalence 3.8 times of those with normal blood pressure. Subjects with family history of type II diabetes had a prevalence 2.0 times of no family history, and women who had delivered babies over 4 kg had a prevalence 1.9 times of those who had babies less than 4 kg. Subjects with waist/hip ratio (WHR) >or= 1.0 had a prevalence 5.1 times of those with WHR < 1.0. The prevalence of type 2 Diabetes was higher with increasing amount of cigarette smoking and the duration (in years) of smoking or increasing drinking (alcohol) and the duration (in years) of drinking. CONCLUSIONS: The blood glucose level was found to be positively correlated with TC or TG, and negatively correlated with HDL-ch. This study demonstrated again that TC, TG, HDL-ch, obesity, hypertension, family history of type 2 diabetes and in women, delivery of babies over 4 kg and age were risk factors of type 2 diabetes. PMID- 11860939 TI - [The expression of P16, P21(WAF1/CIP1) and Cyclin D1 proteins in skin of patients with arseniasis caused by coal-burning and their significance]. AB - OBJECTIVE: To study the relationships between arseniasis and skin carcinoma. METHODS: The expressions of p16, p21(WAF1/CIP1) and Cyclin D1 proteins in skin of arsenic intoxication patients caused by coal-burning was measured by immunohistochemistry methods. RESULTS: The intensity and density, and the positive percentage of p16 protein were gradually reduced as the skin pathological changes progressed. Expressions of p16 protein was lower in the carcinoma group (A group), pre-carcinoma group (B group) and overkeratosis group (C group) than the general cell proliferation group (D group) and normal group (P < 0.05, P < 0.01). On the other hand, the cell density and the positive percentage of p21(WAF1/CIP1) and Cyclin D1 proteins were gradually increased as the skin pathological changes progressed. The expression of p21(WAF1/CIP1) protein in all pathological groups were higher than the normal group (P < 0.05, P < 0.01), and expression of CyclinD1 protein in A, B and C groups were higher than the D group and normal group (P < 0.05, P < 0.01). CONCLUSIONS: Skin overkeratosis may be the earlier stage of skin carcinoma, lower expression of p16 protein and overexpression of Cyclin D1 protein may be considered as prognostic biomarkers to skin carcinogenesis; p16, p21(WAF1/CIP1) and Cyclin D1 may play important role in the cooperative way in the development of arseniasis caused by coal-burning and further progression to skin carcinoma. Monitoring of the expression of p16, p21(WAF1/CIP1) and Cyclin D1 proteins may be valuable for early detection and early treatment and prognostic assessment of skin carcinoma caused by coal-burning. PMID- 11860940 TI - [Effects of fluoride on the expression of c-fos and c-jun genes and cell proliferation of rat osteoblasts]. AB - OBJECTIVE: To investigate the effects of sodium fluoride (NaF) on the expression of c-Fos and c-Jun genes and osteoblast cell proliferation of rat osteoblasts. METHODS: Osteoblastic cells were isolated from baby rat calvaria, and cultured in the presence of different doses of NaF (10(-5) mol/L, 10(-4) mol/L, and 10(-3) mol/L). Cell proliferation was measured by the MTT method, and c-Fos and c-Jun expression was detected by the immunohistochemistry combined with image analysis compute system. RESULTS: RESULTS: from the MTT assay showed that NaF increased the proliferation of rat osteoblast and induced the expression of c-Fos and c-Jun genes. The increases of c-Fos and c-Jun expression by the 3 different doses of NaF were 5.4%, 15.4% and 42.3% for the c-Fos gene and 12.1%, 14.4% and 38.6% for c-Jun gene respectively (P < 0.05 and P < 0.01). CONCLUSION: NaF increased rat osteoblastic cell proliferation and the expression of genes involved in cell proliferation. PMID- 11860941 TI - [Influence of free radical inducer on the level of oxidative stress in brain of rats with fluorosis]. AB - OBJECTIVE: To study changes in content of lipid peroxide and composition of fatty acids in the brain of rats affiliated with chronic fluorosis after treatment with free radical inducer (ferric ion). METHODS: Thirty-six Wistar rats were divided into three groups, fed with similar fodder and varied concentrations of fluoride in drinking water, and were killed five months after treatment. Lipid peroxidation was induced by ferric ions. Malondialdehyde content in brain was analysed by high-performance liquid chromatography; oxygen consumption was determined with an oxygen electrode and fatty acid composition was measured by gas chromatography in brain tissues of the rats. RESULTS: In the brain tissues, content of malondialdehyde and oxygen consumption increased, composition of polyunsaturated fatty acids decreased and that of saturated fatty acids decreased after treatment with free radical inducer in the treated group, as compared with those in control group. CONCLUSION: Over uptake of fluoride for a long term could cause potential increase in the level of oxidative stress in the brain tissue. PMID- 11860942 TI - [Effects of different catechins on cell cycle arrest and induction of apoptosis in LoVo cells]. AB - OBJECTIVE: To investigate the effects of green tea (-)-epigallocatechin-3-gallate (EGCG), (-)-epigallocatechin (EGC), (-)-epicatechin-3-gallate (ECG)and (-) epicatechin (EC) on the cell cycle arrest and induction of apoptosis in human colon carcinoma LoVo cell line. METHODS: The cytotoxicity, cell cycle arrest and apoptosis-associated alterations in morphology and biochemistry were examined using the tetrazolium salt reduction (MTT) assay, gel electrophoresis, transmission electron microscope and flow cytometry after treatment of LoVo cells with the four catechins. RESULTS: Treatment of LoVo cells with EGCG and EGC resulted in the growth suppression and induction of apoptosis in a time- and concentration-dependent manner. ECG and EC, however, did not have the same effects. In addition, the four catechins had different effects on the cell cycle of LoVo cells. Treatment with EGCG, EGC and ECG caused LoVo cells arrest at G1 phase in the cell cycle progression, whereas EC resulted in an arrest at S phase. CONCLUSIONS: The cytotoxicity of the four catechins to LoVo cells is significantly different. The difference is associated with their effects on cell cycle and induction of apoptosis in LoVo cells. PMID- 11860943 TI - [The prevention of primary liver cancer by selenium in high risk populations]. AB - OBJECTIVE: To study the preventive effects of selenium on primary liver cancer. METHODS: After screening of blood samples in 18,000 males from 20 to 65 years-old in Qidong, Jiangsu province (a high risk area for liver cancer), 2,065 cases of HBsAg positive, AFP negative and normal liver function (normal ALT values) were found. The subjects were randomly divided into two groups, based on their residence areas; 1,112 subjects (experimental group) received one tablet of sodium selenite (0.5 mg Se) every day and 953 subjects (control group) received one placebo tablet every day. RESULTS: During three years of intervention and follow up, the blood selenium concentration and glutathione peroxidase activity of the subjects in the experimental group were increased and had significant difference as compared with those of the control group (P < 0.01). At the same time, the prevalence rate of micronucleus cells in peripheral blood lymphocytes in the experimental group was significantly lower than that of the control group (P < 0.01), and the incidence of new liver cancer in the experimental group (3 057.55/10(6), 34 cases out of 1,112 subjects) was significantly lower than the control group (5 981.11/10(6); 57 cases out of 953 subjects) (P < 0.01). CONCLUSION: The results confirms that selenium supplementation in general populations lived in high risk is effective in the prevention of liver cancer and the using of selenium tablets is simple and feasible. PMID- 11860944 TI - [Studies on cell transformation and cell cycle in human embryo lungs induced by nickel compounds]. AB - OBJECTIVE: To elucidate the association between nickel compounds and occupational lung cancer. METHODS: Human embryo lung cells (MRC-9 and IMR-90) were transformed by water-insoluble nickel oxide and nickel subsulfide, and changes in cell cycle of these transformed cells were analyzed by flow cytometric technique. RESULTS: Morphological transformation in MRC-9 and IMR-90 cells were induced after exposure to nickel compounds for 3 - 5 months, including formation of transforming foci, overlaying growth and epithelium-like changes in cells, etc. Some transformed cells could form colonies in soft agar media. Changes in cell cycle of the transformed cells were observed with flow cytometry. Proportions of cells at G(1) phase and G(2)/M phase were 62.61% and 16.37%, respectively, in the untreated MRC-9 cells and hyperploid cells were hardly seen in cell cycle plots. But, in the transformed MRC-9 cells, proportion of cells at G(1) phase reduced to about 50%, and that at G(2)/M phase increased to over 20% and that of hyperploid cells to over 10%. The same phenomenon was observed in the IMR-90 cells. CONCLUSIONS: Water-insoluble nickel compounds can induce morphological transformation of human embryo lung cells and those transformed cells appeared certain biological characteristics of tumor cells. PMID- 11860945 TI - [A study on apoptosis and activity of SAPK/JNK in adrenocortical cells induced by cadmium chloride]. AB - OBJECTIVE: To study apoptosis induced by cadmium chloride and the alteration in activity of stress activated protein kinase (SAPK) in adrenocortical cells. METHODS: Fasciculata-glomerulosa (FG) cells of male guinea pigs were dispersed and primarily cultured in vitro. Features of apoptotic cells were observed using combined labeling with annexin-V and propidium iodide (PI) and flow cytometry, and activity of SAPK was determined with immunoprecipitation and chemiluminescence assay. RESULTS: Apoptosis rate of FG cells increased with dose of cadmium chloride (CdCl(2)) two hours after treatment with 6.25 - 200.00 micromol/L of it. The average percentages of apoptotic cells ranged from 9.90% to 82.35%, with significant difference in the groups treated with 25, 50, 100 and 200 micromol/L of CdCl(2), as compared with the control group (P < 0.01). Regression analysis showed that occurrence of apoptosis correlated with the dose of cadmium chloride in a dose-response pattern. In the meanwhile, there were obviously elevated percentages of apoptotic cells as the increase in duration of incubation, ranging from 5.58% to 73.08% for incubating cells with 50.00 micromol/L of CdCl(2), from 15 minutes to four hours. Duration of incubating cells with cadmium chloride correlated to occurrence of apoptosis in a time effect manner. There were no remarkable changes in the activity of SAPK in adrenocortical cells two hours after incubation with 25.00 - 100.00 micromol/L of CdCl(2). CONCLUSIONS: Apoptosis can be induced by cadmium chloride in adrenocortical cells and its mechanism should be studied further. PMID- 11860946 TI - [Effects of ob genotype on early development in mice]. AB - OBJECTIVE: To study the effects and the extent of dominance of ob gene on early development in mice. METHODS: ob genotypes, wild type (+/+), hybrid type (ob/+) and mutant type (ob/ob), were detected by polymerase chain reaction and restrict endonuclease technique. Body weight of mice was recorded daily for the first seventh weeks of life. Their wet weight of white adipose tissue (WAT) and brown adipose tissue (BAT) was compared at the 7th, 14th and 24th days after birth. RESULTS: Body weight of ob/ob mice increased rapidly two weeks after birth and reached two times of that of +/+ and ob/+ mice seven weeks after birth. WAT of ob/ob mice was significantly higher than that of +/+ and ob/+ mice 24 days after birth. Effect of ob genotype on BAT was earlier than that on WAT. Body weight and WAT of ob/+ mice ranged between those of +/+ and ob/ob mice. Dominance index, reflecting the effect of ob gene on body weight, ranged from 1.0 - 0.5 within the first two weeks after birth, appearing co-dominance. And, dominance index approximated to 0.0 six weeks after birth, appearing almost complete recession. CONCLUSION: Effect of ob gene on body weight and WAT initiated two weeks after birth and increased with age, developing gradually from co-dominance to recession in their early life. PMID- 11860947 TI - [A preliminary study on body level of nitric oxide in workers exposed to carbon disulfide]. AB - OBJECTIVE: To study changes in body plasma level of nitric oxide (NO) in workers exposed to carbon disulfide (CS(2)) and its possible mechanism. METHODS: Plasma levels of NO and lipid peroxide (LPO) and activity of erythrocyte superoxide dismutase (Ery-SOD) were determined in the workers exposed and unexposed to CS(2) in a chemical synthetic fiber works. RESULTS: Level of NO was (43.28 +/- 19.83) and (50.07 +/- 21.01) micromol/L in high- and low-dose exposed groups, respectively, significantly lower than that in the control group, which was (70.66 +/- 26.83) micromol/L (P < 0.05). Activity of Ery-SOD was 4 832.21 u/g Hb and 3,520.80 u/g Hb in high- and low-dose exposed groups, respectively, significantly higher than that in the control group, which was 2,425.34 u/g Hb (P < 0.05). Level of LPO was 19.38 and 17.09 micromol/L in high- and low-dose exposed groups, respectively, significantly higher than that in the control group, which was 4.37 micromol/L (P < 0.05). CONCLUSION: Occupational long-term exposure to CS(2) could induce reduction of NO level in the body with its mechanism related to increase of super-oxygen cation O(2)(*-) induced by CS(2). PMID- 11860948 TI - [The changes in serum antibody level after immunization with HFRS vaccine]. AB - OBJECTIVES: To observe the changes in serum antibody level after mass immunization with vaccine against hemorrhagic fever with renal syndrome (HFRS) and to evaluate its efficacy and effectiveness in the prevalent areas. METHODS: Healthy people aged 16 to 60 years in the villages were recruited as study subjects, excluding those suffered from HFRS previously, going out for more than nine months and those with contraindications, and were randomly allocated into immunization and control groups with 10,460 and 16,159 persons, respectively. Specific IgG antibody was determined with indirect immunofluorescent assay (IFA) and neutralizing antibody (NA) was determined with micro CPE method. RESULTS: Two weeks after the full-course immunization, sero-conversion rate for IFA reached 100% in those sero-negative before immunization, with a 95% confidence interval of 96.3 - 100.0%, and that for NA 44.4%, with a 95% CI of 22.0% - 69.0%. Geometric mean titer (GMT) were 72.1 and 4.6 for IFA and NA, respectively. Booster immunization was provided for them one year later. Positivity of IFA and NA was 28.6%, 83.3%, 75.0%, 53.1%, 22.6% and 14.8%, 55.6%, 35.0%, 31.3%, 26.0%, before booster immunization, two weeks, one year, one and a half years, two years after booster immunization, respectively. CONCLUSION: HFRS vaccine had good immunogenicity, but its duration of serum antibody sustenance was relatively short. PMID- 11860949 TI - [Analysis of hepatitis G virus genotypes in some areas of China]. AB - OBJECTIVE: To study the genotype distribution of hepatitis G virus (HGV) isolated from different population in some areas of China. METHODS: A total of 12 strains of HGV were isolated from different areas of China (three from Guangxi Zhuang Autonomous Region, three from Hebei Province, one from Heilongjiang, Jiangxi, Guangdong, Jiangsu, Anhui, Jilin Provinces and Xinjiang Autonomous Region each). The HGV 5' untranslated region (UTR) of the 12 isolates was sequenced and compared with other 26 isolates previously reported (17 from China and nine from other countries). RESULTS: The nucleotide sequences of five HGV clones isolated from the same patient with HGV infection were identical, with the homology of 99.1% - 100%, indicating that variation of HGV in the same human body was very limited. Genotyping could be based on the sequence of one positive clone isolated from a patient with HGV infection. The homology of the nucleotide sequences in the 12 isolates was 96.8% - 99.4%, suggesting that they belonged to the same genotype. The homology of the 12 isolates was 88.0% - 89.7%, 87.2% - 90.3% and 94.7% - 96.3%, respectively, as compared with HGV genotypes 1 (U36380), 2 (U44402) and 3 (D90601), which indicated that all the 12 HGV isolates belonged to genotype 3. Phylogenetic distance analysis of the 38 isolates suggested that HGV strains isolated from China could be further divided into different subgroups. CONCLUSION: The majority of Chinese HGV isolates belonged to genotype 3 and they could be further divided into different subgroups. PMID- 11860950 TI - [Pathogenecity and replication of transfusion transmitted virus in liver tissue by in situ hybridization]. AB - OBJECTIVE: To study its pathogenecity based on infection status and replication of transfusion transmitted virus (TTV) in the liver tissues. METHODS: Using recombinant plasmid as template and symmetrical and asymmetrical polymerase chain reaction (PCR), double-strand and single-strand TTV probes were labeled with digoxigenin. Fifty-six cases of liver biopsy specimens were detected by in situ hybridization, and six positive cases of them were tested further with single strand probes. RESULTS: TTV DNA was found in 14 of 51 patients with non A and non G hepatitis (27.5%), and five cases in the control group were negative. TTV DNA was mainly observed in the nuclei of hepatocytes and pathologic changes and necrosis in the infected liver cells were not so apparent, but they could be seen in the liver tissues of acute, chronic and severe hepatitis, as well as of liver cirrhosis. TTV genome strand hybridization detected by single-strand probe was consistent with that by double-strand probe. Both TTV genome strand and its complementary strand were present in the two cases of liver specimens, but hybridizational signals were weaker in the latter than those in the former. CONCLUSION: TTV could cause injury and replicate itself in the liver, but no obvious and direct injury to hepatocytes. PMID- 11860951 TI - [An epidemiological study on pesticide poisoning with mixed preparation]. AB - OBJECTIVE: To investigate the incidence of pesticide poisoning with mixed preparation and risk factors contributing to it. METHODS: Twenty-five administrative villages were studied in three townships of Jiangsu and Shandong Provinces with cluster sampling. The sampled villages were divided into study group (12 villages) and control group (13 villages) with approximately equal number of people each. Pesticide sprayers in the study group applied mixed preparation containing organophosphorous and dimethrin and those in the control group applied single organophosphorous preparation. All the sprayers in sampled villages were interviewed with a same questionnaire about their situation of pesticide spraying during the observation period. Cases of pesticide poisoning were ascertained by seniors physicians in occupational diseases after group discussion. RESULTS: Incidence of pesticide poisoning was higher in 2 179 sprayers of the study group (10.10 %) than that in 2 615 of the control group (2.29%), with a highly statistically difference (chi(2) = 12.46, P < 0.01) and an risk ratio (RR) of 4.45 (95% CI = 1.94 approximately 10.05) adjusted for other risk factors by logistic regression. Other risk factors for pesticide poisoning with mixed preparation were smoking or taking food during spraying, leaking or breakdown of sprayers, without washing their whole body as soon as possible after spraying, poor personal protection, spraying for a long time, spraying by women and spraying pesticide on cotton. CONCLUSIONS: If toxicity of mixed preparation of single pesticide showed a synergistic or additional effect, risk of poisoning by spraying mixed preparation increased. Health education and technical training should be strengthened for pesticide sprayers. PMID- 11860952 TI - [The gene polymorphism of delta-aminolevulinate dehydratase (ALAD) in 530 cases of Chinese Han population]. AB - OBJECTIVE: To explore the distribution of allele and genotype frequencies of delta-aminolevulinate dehydratase (ALAD) gene in Chinese Han population. METHODS: Blood specimens were collected from 530 unrelated Han individuals in the Central Plain of China. The ALAD genotype was determined using polymerase chain reaction restriction fragment length polyporphism (PCR-RFLP) method. RESULTS: The frequencies of genotype in Chinese Han ethnic were 93.58% (496) for the ALAD(1 - 1) homozygote, 6.23% (33) for the ALAD(1 - 2) heterozygotes and 0.19% (1) for the ALAD(2 - 2) homozygote, respectively. The allele frequencies of the ALAD(1) and ALAD(2) were 0.967 and 0.033, respectively. CONCLUSION: The frequency of the ALAD(2) allele in Chinese Han population was similar to that in Asian populations and lower than that in Caucasian population. PMID- 11860954 TI - [Study on expression of P53mt protein in skin of patient with arseniasis caused by coal-burning]. AB - OBJECTIVE: To study expression of P53mt protein in skin of patient with arseniasis caused by coal-burning. METHODS: According to pathological diagnosis, the patients were divided into carcinoma group (A group, 18 cases), pre-carcinoma group (B group 11 cases) and general pathological change group (C group, 39 cases). Immuno-histochemical technique of two steps was used to detect expression of P53 mt protein. RESULTS: Among A, B and C groups, the positive cell densities of P53mt protein were 38.07 +/- 29.00, 17.16 +/- 15.00 and 4.05 +/- 8.24 respectively, but no positive cell in normal group; there was statistical significance among A, B and C groups (A and C: P < 0.01, A and B: P < 0.01, B and C: P < 0.05); positive percentages of A, B and C groups were 88.89%, 72.73% and 25.64% (A and C: P < 0.01; B and C: P < 0.05), respectively, for difference of grade constitution, there was statistical significance (P < 0.01) among three groups; grade II and III made up 38.89% in A group, but 9.09% in B group and 2.56% in C group. CONCLUSIONS: P53mt gene play a certain role in the development of skin cancer from arseniasis caused by coal-burning; the detection of P53mt protein can show whether pathological changes of skin due to the disease is malignant or benign. PMID- 11860953 TI - [The inhibitory effect of CLA on mice forestomach neoplasia induced by B(a)P]. AB - OBJECTIVE: To study the inhibitory effect of CLA on the postinitiation phase of forestomach neoplasia formation induced by B(a)P and explore its possible mechanism of anticarcinogenesis. METHODS: Kunming mice were divided into five groups, i.e. salad oil control group, CLA control group, B(a)P group, B(a)P + high dose CLA group and B(a)P + low dose CLA group. The experimental period was 23 weeks, and the endpoints included cell proliferation, expression of Pan-ras P21 and some enzymes. RESULTS: Short term CLA treated significantly inhibited the forestomach neoplasia formation induced by B(a)P at postinitiation phase. The incidence of tumor in the groups of B(a)P, B(a)P + high dose CLA and B(a)P + low dose CLA was 100%, 60% and 69% respectively (P < 0.05). The possible mechanism of this anticarcinogenic effect may be related to the inhibition of cell proliferation and the induction of the activities of GSH-Px, GST and SOD, irrespective of the regulation of expression of Pan-ras P21. CONCLUSION: CLA is a promising chemopreventive agent, and its anticarcinogenic effect may involve its effects on the redox system. PMID- 11860955 TI - [Assessment of quality of life in patients with diabetes mellitus and impaired glucose tolerance]. AB - OBJECTIVES: To understand the quality of life (QOL) in patients with diabetes mellitus and impaired glucose tolerance, and investigate factors affecting their QOL. METHODS: Using the SF-36 instrument to assess QOL among 108 patients with diabetes mellitus, 109 patients with impaired glucose tolerance and 116 normal glucose tolerance subjects. RESULTS: In subjects with diabetes mellitus, the proportion of general perceived health assessed to be excellent or good was 12.04%; as compared with the same-age people, the proportion assessed to be good or fairly good was 62.04%. In subjects with impaired glucose tolerance, the two proportions was 13.76% and 69.72% respectively. The total score attained excellent or good level was 72.23% and 83.49% respectively in patients with diabetes mellitus and impaired glucose tolerance. The mean score of multi-item dimensions assessment (ranged from 58.33 to 87.38) decreased in diabetes mellitus; the lowest score being emotional role functioning and the highest mean score being physical functioning. Compared with normal glucose tolerance subjects, the mean score of physical functioning, physical role functioning, general health perception, vitality and the total score in diabetes mellitus was significantly decreased; and compared with in impaired glucose tolerance patients, the mean score of vitality, mental health and the total score was significantly decreased in diabetes mellitus. The result of correlation analysis demonstrated that age, occupational, duration of disease, number of symptoms and complications, level of fasting blood glucose and 2-hour past oral blood glucose affected the QOL of diabetes mellitus patients. CONCLUSIONS: The results suggested that to enhance the QOL in patients with diabetes mellitus, control of plasma glucose, pay attention to psychological treatment and measures related to diabetes mellitus with different characteristics must be early adopted. PMID- 11860956 TI - [Attitude to voluntary HIV testing and result disclosure among rural residents in China]. AB - OBJECTIVE: To identify the determinants of individual acceptance of voluntary HIV testing and the scope of test results disclosure. METHODS: A cross-sectional study was performed to interview 1 057 subjects aged 15 - 49. RESULTS: Among 653 subjects who had heard of HIV/AIDS, 575 (88.1%) reported willingness to be tested for HIV infection. Of subjects who would accept the offer, 90.2% agreed to disclose testing results to their spouse, 77.1% to other family member, 54.8% to their friends and 50.8% to their neighbors. Logistic regression analysis indicated that the acceptability was associated with being a man, and having better knowledge in HIV/AIDS. CONCLUSIONS: The level of acceptability of voluntary HIV testing is high among people who know about HIV/AIDS. Given the large number of people in rural China who never heard of HIV/AIDS, it is important to carry out information-oriented education activities to increase AIDS awareness among rural residents. PMID- 11860957 TI - [Studies on the spatial and familial aggregation of double Y nuclear family in gastric cancer pedigree]. AB - OBJECTIVE: To explore whether there existed spatial and familial aggregation in gastric cancer in Yangzhong City of Jiangsu Province. METHODS: Using the data collected from cancer incidence registry and report system in Yangzhong City and newly designed investigation for double Y nuclear family, a genetic epidemiological study was conducted in 448 nuclear pedigrees with gastric cancer and 437 control pedigrees, involving 5 242 family members. RESULTS: Chi square test of goodness-of-fit for Poisson distribution showed there was difference between real incidence rate of gastric cancer in all townships and towns in Yangzhong and expected distribution, with chi(2) = 191.07, P < 0.001. The highest rate was in Sanyao, Changhong and Youfang townships, which was 2.2 times higher as that in the areas with low-incidence rate. Prevalence rate of gastric cancer was significantly higher in the siblings (5.42%) and parents (8.65%) of the probands, than that in siblings and parents of their spouses (4.94% and 3.20%, respectively). Prevalence rate of gastric cancer in the offspring with both parents suffered from gastric cancer was the highest, 22.58%. Test for goodness of-fit of binominal distribution showed that there was significant difference between observed number and expected number of pedigrees with gastric cancer, with P < 0.0001. CONCLUSION: There was not only significantly spatial aggregation of gastric cancer in Yangzhong City, but also familial aggregation, which coincided with a mode of multi-genetic inheritance based on preliminary results and laid a foundation for exploring the difference in gentic susceptibilities and environmental factors for gastric cancer in Yangzhong City. PMID- 11860958 TI - [A survey on physical development in children of 7 nationality minorities in China]. AB - OBJECTIVE: To obtain the data on physical development in minority children from 0 to 14 years of age in China. METHODS: Totally 41 350 children of Korean, Mongolian, Li, Miao, Puyi, Tung and Tibetan were investigated with stratified random sampling. RESULTS: With height and weight as indicators, physical development in Korean and Mongolian children was better, that in Tibetan and Miao medium, and that in Buyi and Tung lower. Height in Li children was medium, but their weight was lower. Mongolian children ranked the highest in Kaup index and Ververck index with a sturdy physical status and Li children was the lowest in that with a leaner body. CONCLUSIONS: There was a very significant difference in physical growth and development among children of seven nationality minorities investigated. But, difference in their resident areas should be considered in analysis for such a difference between nationalities. PMID- 11860959 TI - [A study on relationship between blood lead level in children and fetal umbilical cord]. AB - OBJECTIVE: To study the relationship between blood lead level in children and fetus. METHODS: Blood specimens were collected from the fingers of children aged one to five years and from umbilical cord when the babies were born, to determine their lead level with atomic absorption spectrophotometry. RESULTS: The average blood lead level was (0.42 +/- 0.15) micromol/L in 1 122 children. Blood lead level in 27.54% of the children was above >/= 0.483 micromol/L. The average umbilical blood lead level was (0.16 +/- 0.12) micromol/L in 127 neonates. Blood lead level in 0.79% of the new-born babies were greater than >/= 0.483 micromol/L. CONCLUSION: Higher blood lead level in children was mainly caused by environmental pollution and unhealthy life behaviors. Background blood lead in fetuses has already reached a higher level, so prevention from lead poisoning in children should begin during pregnancy. PMID- 11860960 TI - [The relationship between cadmium-induced inhibition of splenic lymphocyte function and cell apoptosis]. AB - OBJECTIVE: To explore the relationship between cadmium-induced inhibition of splenic lymphocyte function and cell apoptosis in vitro. METHODS: The splenic lymphocytes of mice were exposed to 3.10, 6.25, 12.50, 25.00, 50.00 micromol/L of cadmiun chloride (CdCl(2)) for various time period, to measure the lymphocyte transformation by MTT colorimetric assay and to detect the apoptosis in splenocytes by DNA agarose electrophoresis and flow cytometry (FACS). RESULTS: CdCl(2) could significantly inhibit the function of lymphocyte transformation in a dose-response pattern at concentrations of 25.00 and 50.00 micromol/L, with inhibition rates of 50% and 78% in ConA-stimulating group and 39% and 55% in LPS stimulating group, respectively. CdCl(2) could also induce apoptosis of splenic cells at concentrations of 12.50 to 50.00 micromol/L. RESULTS: of FACS analysis showed 30% - 60% of cell apoptosis. Higher concentration of CdCl(2) could also cause reduction of cell survival. Effect of Cd-induced apoptosis was earlier and at a lower concentration of CdCl(2) than that of Cd-induced inhibition of lymphocyte transformation. CONCLUSION: CdCl(2) could induce cell apoptosis in vitro, which was one of the mechanisms of its suppression of lymphocyte function. PMID- 11860961 TI - [A study on mechanism for cytotoxicity of nitrobenzene to hepatocarcinoma cell line]. AB - OBJECTIVE: To study the mechanism for cytotoxicity of nitrobenzene to hepatocarcinoma cell line. METHODS: Cytotoxicity was determined with MTT assay in SMMC-7721 cell strain, human hepatocarcinoma cell, incubated with nitrobenzene at varied concentrations in vitro for 24 hours. The effect of an agent to clear reactive oxygen on protection from cell death was observed. Cell cycle of hepatocytes was also observed using flow cytometry and their DNA damage was detected with single-cell gel electrophoresis assay. RESULTS: Nitrobenzene at concentration of 8 mmol/L or over could induce death of hepatocytes obviously with a survival rate of less than 82%, and addition of superoxide dismutase, hydrogen peroxidase or mannitol to the incubation media could inhibit its cytotoxicity partly, with survival rates of 93.7%, 96.6% and 93.2%, respectively. Nitrobenzene could affect cell cycle of hepatocytes, showing a decreased fraction of G(1) phase and increased fraction of S and G(2)/M phase, but could not induce DNA strand breaks in them. CONCLUSIONS: Nitrobenzene had cytotoxicity to hepatocytes cultured in vitro, in which reactive oxygen species may play a certain role. Nitrobenzene was a non-genotoxicant and could cause cancer by promotion of cell proliferation. PMID- 11860962 TI - [Effect of inorganic arsenic on gap junctional intercellular communication between human skin fibroblasts]. AB - OBJECTIVE: The aim of this study is to investigate the effect of inorganic arsenic on gap junctional intercellular communication (GJIC) between human skin fibroblasts. METHODS: GJIC between human skin fibroblasts was detected by the scrape loading/dye transfer assay. RESULTS: Arsenite at concentrations from 0.005 micromol/L to 5.0 micromol/L significantly inhibited the dye transfer between human skin fibroblasts in a dose dependent manner. The dye transfer between human skin fibroblasts was also inhibited by arsenate, while the dose-dependent effects were not observed. Further study found that arsenite at concentrations of 0.5 micromol/L and 5.0 micromol/L significantly increased protein kinase C (PKC) activities in cells, especially in the membrane fraction. CONCLUSIONS: Arsenite and arsenate significantly inhibited GJIC between human skin fibroblasts, suggesting that inorganic arsenic may play an important role in the process of tumor promotion. Furthermore, PKC activation may be involved in the mechanisms of GJIC inhibition by inorganic arsenic. PMID- 11860963 TI - [The effect of alcohol on c-fos gene expression in rat embryo neuroglial]. AB - OBJECTIVE: This paper is aimed to explore the mechanisms of brain development abnormality induced by alcohol. METHODS: Astrocytes and oligodendrocytes of 19 day rat embryo were exploited and cultured in vitro, and alcohol and its metabolite product (acetaldehyde) were added to DMEMF(12) medium. After different exposure times, c-fos expression of astrocytes and oligodendrocytes was measured by the immunocytochemistry technique. RESULTS: Changes in c-fos gene expression induced by alcohol and acetaldehyde was time and dose dependent. After 1 hr exposure, alcohol and acetaldehyde affected c-fos gene expression in two kinds of neuralglia. C-fos positive expression reached peak value after 2 hr, but recovered after 72 hr and showed special time phase expression. CONCLUSIONS: Alcohol and acetaldehyde cause abnormal increase of c-fos gene expression in astrocytes and oligodendrocytes. This abnormal expression may play an important role in abnormal brain development induced by alcohol. PMID- 11860964 TI - [Change of NOS activity in hypoxia and cold-induced blood vessels damage and its biological significance]. AB - OBJECTIVES: To explore the therapeutic methods for hypoxia and cold induced blood vessel damage by observing the changes of NO synthase (NOS) activity in hypoxia and development of coldm induced blood vessels damage, and studying the effects of SOD and vitamin C on the changes of NOS activity. METHODS: The main artery was isolated from Wistar rats and exposed to hypoxia and/or cold environment in vitro. LDH activity was measured using an automatic biochemical analyzer and NOS and SOD activities in blood vessels were analyzed respectively by Griess assay and adrenaline autoxidantion method. RESULTS: NOS activity in hypoxia, cold and hypoxia-cold damaged vessels decreased by 18.2%, 19.1% and 25.9% respectively after exposure as compared with the control vessels. The changes of NOS activity were time dependent. The NOS activity in damaged vessels was negatively correlated with LDH activity in the culture medium of the respective vessels. The SOD activity in hypoxia and/or cold damaged vessels also decreased significantly. Giving SOD (200 u/ml) or vitamin C (50 mg/ml) immediately after hypoxia and/or cold exposure increased the NOS activity and SOD activity in the damaged vessels significantly, as compared with the non-treatment controls. CONCLUSIONS: The decrease of NOS activity is closely related to the development of damage induced by hypoxia and/or cold injury. SOD and vitamin C could protect the blood vessels exposed to hypoxia and/or cold from damage, possibly by increasing NOS activity. PMID- 11860965 TI - Keratocyte density of central human cornea after laser in situ keratomileusis. AB - PURPOSE: To determine changes in keratocyte density in the first year after laser in situ keratomileusis (LASIK). DESIGN: Prospective interventional cohort study. METHODS: Seventeen eyes of 11 patients received LASIK with a planned 180-microm flap to correct refractive errors between -2.0 diopters and -11.0 diopters. Images of the full-thickness cornea were obtained by using confocal microscopy in vivo before LASIK and at 1 week, 1, 3, 6, and 12 months after LASIK. Bright objects (that resembled keratocytes) in images without motion blur were manually counted by one observer. Cell densities were determined in anterior and posterior halves of the stromal flap, anterior and posterior halves of the layer 100 microm thick immediately deep to the ablation (retroablation layer), and in the posterior third of the stroma. The region of stroma that was ablated (as measured 1 month after LASIK) was omitted from the preoperative analysis. RESULTS: Keratocyte density in the anterior flap was 28,978 +/- 5849 cells/mm(3) (mean +/- SD) pre-LASIK, and was decreased at all postoperative examinations, but the difference was not significant until 12 months after LASIK (22% decrease). Keratocyte densities in the posterior flap were 20,397 +/- 4215 cells/mm(3) pre LASIK and were decreased by 20%-40% at all postoperative examinations 1 week to 1 year after LASIK. Keratocyte densities in the anterior half of the retroablation layer were 16,605 +/- 3595 cells/mm(3) pre-LASIK and decreased by 16%-30% between 3 and 12 months after LASIK. Keratocyte densities in the posterior half of the retroablation layer and posterior stroma did not change. CONCLUSIONS: Keratocyte densities in the posterior flap and anterior retroablation layer (regions adjacent to the lamellar cut) decrease at 1 week and 3 months, respectively, after LASIK and remain decreased in these regions at 12 months after LASIK. In the anterior flap, keratocyte density decreases 1 year after LASIK. The long-term effects of these cellular deficits, if any, require further study. PMID- 11860966 TI - The measurement of corneal epithelial thickness in response to hypoxia using optical coherence tomography. AB - PURPOSE: To determine if corneal epithelial thickness increases in association with corneal edema induced by wearing soft contact lenses during eye closure. DESIGN: Experimental study. METHODS: One eye (randomly selected) of twenty noncontact lens wearers (10 males and 10 females, age 35.6 +/- 9.6 years) was patched during 3 hours of soft contact lens (SCL) wear and the contralateral eye acted as a control. Corneal and epithelial thickness of both eyes was measured before and after SCL wear using optical coherence tomography (OCT). RESULTS: Immediately after contact lens removal, total corneal thickness was increased significantly by 13.8 +/- 2.3% (mean +/- SD) compared with baseline (P <.0001, paired t test) and after 100 minutes was still 4.5 +/- 2.3% thicker than baseline (P <.0001, paired t test). The control eyes showed no change in total corneal thickness (P >.05, paired t test). Immediately after contact lens removal, corneal epithelial thickness was increased by 1.7 +/- 4.8%, but this change was not statistically significant (P >.05, paired t test). Following contact lens removal, epithelial thickness changed significantly (Repeated measure analysis of variance [Re-ANOVA]: F((7,133)) = 4.91, p(H-F) < 0.001) over the next 100 minutes with thinning recorded at 60, 80, and 100 minutes (P <.05, paired t test). There was no significant change over time in epithelial thickness of the control eyes (Re-ANOVA: F(4, 76) = 0.91, p(H-F) = 0.464). CONCLUSION: OCT demonstrated that corneal epithelial thickness does not increase in response to hypoxia from SCL wear and eye closure, in contrast to a significant increase in total corneal thickness. PMID- 11860967 TI - Analysis of elements of interlenticular opacification. AB - PURPOSE: To report the histopathologic and ultrastructural features of three cases of interlenticular opacification (ILO) between piggyback intraocular lenses. DESIGN: Interventional case series with clinicopathologic correlation. METHODS: Three pairs of acrylic piggyback lenses were explanted due to decrease in visual acuity associated with ILO. Lenses were evaluated with gross and light microscopic examinations in all cases. The anterior lens in one case was examined with scanning electron microscopy and energy dispersive x-ray spectroscopy. RESULTS: The material opacifying the interlenticular space was composed mostly of retained/regenerative cortical material in all cases. From the peripheral interface towards the central interface, the opacifying material changed as the interlenticular space was progressively narrower. The material attached to the peripheral interface, where the interlenticular space was wider, was very thick. At the midperipheral interface, the thick cortical material was broken into multiple globules due to liquefactive degeneration. At the paracentral zone, compression of the globules formed a flat, compact layer of an amorphous material. At the central interface (contact zone), almost no material could be found between the piggyback lenses. CONCLUSIONS: Analyses of ILO cases where all the components of the opacifying material were in situ allowed us to confirm that the pathogenesis of this complication is similar to that of posterior capsule opacification; thus, careful removal of lens epithelial cells and cortical material is mandatory in piggyback implantation. PMID- 11860968 TI - Sensitivity and specificity of frequency-doubling technology, tendency-oriented perimetry, and Humphrey Swedish interactive threshold algorithm-fast perimetry in a glaucoma practice. AB - PURPOSE: To evaluate the sensitivity and specificity of the screening mode of the Humphrey-Welch Allyn frequency-doubling technology (FDT), Octopus tendency oriented perimetry (TOP), and the Humphrey Swedish Interactive Threshold Algorithm (SITA)-fast (HSF) in patients with glaucoma. DESIGN: A comparative consecutive case series. METHODS: This was a prospective study which took place in the glaucoma unit of an academic department of ophthalmology. One eye of 70 consecutive glaucoma patients and 28 age-matched normal subjects was studied. Eyes were examined with the program C-20 of FDT, G1-TOP, and 24-2 HSF in one visit and in random order. The gold standard for glaucoma was presence of a typical glaucomatous optic disk appearance on stereoscopic examination, which was judged by a glaucoma expert. The sensitivity and specificity, positive and negative predictive value, and receiver operating characteristic (ROC) curves of two algorithms for the FDT screening test, two algorithms for TOP, and three algorithms for HSF, as defined before the start of this study, were evaluated. The time required for each test was also analyzed. RESULTS: Values for area under the ROC curve ranged from 82.5%-93.9%. The largest area (93.9%) under the ROC curve was obtained with the FDT criteria, defining abnormality as presence of at least one abnormal location. Mean test time was 1.08 +/- 0.28 minutes, 2.31 +/- 0.28 minutes, and 4.14 +/- 0.57 minutes for the FDT, TOP, and HSF, respectively. The difference in testing time was statistically significant (P <.0001). CONCLUSIONS: The C-20 FDT, G1-TOP, and 24-2 HSF appear to be useful tools to diagnose glaucoma. The test C-20 FDT and G1-TOP take approximately 1/4 and 1/2 of the time taken by 24 to 2 HSF. PMID- 11860969 TI - Pupillary response to four concentrations of pilocarpine in normal subjects: application to testing for Adie tonic pupil. AB - PURPOSE: To determine the amount of pupillary constriction to four different concentrations of pilocarpine in normal human subjects and to determine if pupillary constriction correlates with bioavailability of the instilled concentrations. The amount of pupillary constriction to dilute pilocarpine is utilized as a diagnostic test for Adie tonic pupil as distinguished from a normal pupil response. DESIGN: Twenty healthy volunteers had automated binocular infrared pupillography in the dark after instillation of four different concentrations of dilute pilocarpine. Ocular penetration of eye drops was also assessed using 2% fluorescein sodium as a tracer. METHODS: Prospective institutional double-masked study of both eyes of twenty healthy volunteers, ten with brown irides, ten with blue irides, between the ages of 20-40 years. RESULTS: A pilocarpine dose-dependent curve showed decreased pupil size within 15 minutes, peaking at 30-60 minutes. No difference was noted between right and left eyes, iris color, or corneal permeability. CONCLUSIONS: Normal pupils constrict to dilute concentrations of pilocarpine (0.25% or 0.125%), but constrict insignificantly to concentrations of 0.0313% or 0.0625%. Pupil constriction with 0.0625% pilocarpine should distinguish an Adie pupil from normal. This confirms the utility of this simple office diagnostic procedure. PMID- 11860970 TI - Balloon catheter dilation for treatment of persistent nasolacrimal duct obstruction. AB - PURPOSE: To report the outcome of balloon catheter dilation of the lacrimal duct for treatment of children with persistent nasolacrimal duct obstruction after previous surgery. DESIGN: Interventional case series. METHODS: Thirty-two consecutive children with a history of persistent nasolacrimal duct obstruction following previous surgery were treated with nasolacrimal duct probing with balloon catheter dilation of the distal nasolacrimal duct. Patients were excluded from this study if they had a history of facial trauma, systemic disorders that involved the lacrimal system, or nasolacrimal duct cysts. Outcomes were considered excellent if the patient had complete symptomatic resolution of epiphora and dacryocystitis and normal tear drainage on examination, good if the patient had only minimal residual symptoms or a minimally delayed dye disappearance test, fair if the patient had moderate residual symptoms or delayed tear drainage, and poor if there was no improvement. RESULTS: Thirty-two children (20 with bilateral nasolacrimal duct obstruction and 12 with unilateral nasolacrimal duct obstruction) underwent balloon catheter dilation. Overall results were excellent in 9 (28%) patients, good in 15 (47%) patients, fair in 7 (22%) patients, and poor in 1 (3%) patient. Seven patients had undergone more than one previous procedure. Of these patients, 3 had excellent outcomes, 2 had good outcomes, 1 had a fair outcome, and 1 had a poor outcome following balloon catheter dilation. CONCLUSIONS: Balloon catheter dilation is a safe and generally effective treatment for children with persistent symptoms of nasolacrimal duct obstruction following previous surgery. PMID- 11860972 TI - Impact of visual function on computer task accuracy and reaction time in a cohort of patients with age-related macular degeneration. AB - PURPOSE: To investigate the impact of visual function parameters on computer task performance in patients with age-related macular degeneration (AMD). DESIGN: Interventional case series. METHODS: Eighteen patients with visual impairment due to age-related macular degeneration underwent evaluation of visual acuity using the Early Treatment Diabetic Retinopathy Study protocol, contrast sensitivity using a Pelli-Robson chart, binocular simultaneous visual field using the Esterman program on an automated perimeter, and color vision using Farnsworth D 15. Each subject then completed 125 computer icon identification tasks. Relationships between computer task performance (accuracy and speed) and visual function parameters (visual acuity, contrast sensitivity, visual field, and color vision) were analyzed. RESULTS: Visual acuity and contrast sensitivity in the better eye, weighted average contrast sensitivity, and color vision defects are significantly associated with computer task accuracy. Visual acuity in the better eye, weighted average visual acuity, and color vision defects are significantly associated with performance speed. Visual function parameters and clinical features significantly associated with computer task accuracy in a multiple regression model include weighted average contrast sensitivity (P = 0.001), protan color vision defect (P = 0.002), cataract severity in the better-seeing eye (P = 0.036), and geographic atrophy outside the central macula (P = 0.046). Visual function parameters and clinical features significantly associated with computer task speed in a multiple regression model include color vision defects (deutan, P < 0.001; protan, P < 0.001) and gender (P = 0.05). CONCLUSIONS: Among this cohort of patients with AMD, visual acuity, contrast sensitivity, and color vision defects are significant predictors of computer task performance. Visual function parameters of the better eye played a more significant role than parameters of the worse eye, and contrast sensitivity is the most significant predictor of computer task accuracy. PMID- 11860971 TI - Fundus autofluorescence in patients with age-related macular degeneration and high risk of visual loss. AB - PURPOSE: To describe fundus autofluorescence (AF) patterns and their change over time in patients with age-related macular degeneration (AMD) and high risk of visual loss participating in the drusen laser study (DLS). DESIGN: Randomized clinical trial. METHODS: The study population consisted of 29 patients (35 eyes) participating in the DLS, which is a prospective, randomized, controlled clinical trial of prophylactic laser therapy in patients with AMD and high risk of neovascular complications. The intervention consisted of 16 eyes having prophylactic laser and 19 receiving no treatment. The main outcome measures were changes in the distribution of drusen and AF. Patients were reviewed for a median follow-up or 24 months (range 12-36 months). RESULTS: At baseline, four patterns of fundus AF were recognized: focal increased AF (n = 18), reticular AF (n = 3), combined focal and reticular AF (n = 2), and homogeneous AF (n = 12). At last follow-up, fundus AF remained unchanged in 15 untreated (78%) and in seven treated (43%) eyes. In only one untreated eye, focal areas of increased AF returned to background levels and were no longer detectable at last follow-up, compared with six treated eyes. This difference was statistically significant (P =.03). Only large foveal soft drusen (drusenoid pigment epithelium detachments) consistently corresponded with focal changes in AF, whereas no obvious correspondence was found between small soft drusen located elsewhere and changes in AF. CONCLUSION: The lack of obvious correspondence between the distribution of drusen and of AF found in this study appears to indicate that drusen and AF represent independent measures of aging in the posterior pole. PMID- 11860973 TI - Surgical management of epiretinal membrane in young patients. AB - PURPOSE: To report the outcome of epiretinal membrane (ERM) surgery in young patients. DESIGN: Interventional case series study. METHOD: A retrospective review of 20 young patients who had undergone vitrectomy and epiretinal membrane removal. All patients had a preoperative examination, including fundus biomicroscopy, red free photographs, and fluorescein angiography. RESULTS: Nine patients were males and 11 females (age: 7-26 years; mean: 16.3 years). Follow up ranged from 4-96 months (mean: 21.2 months). The 20 patients comprised 13 cases of idiopathic ERM, six cases in which ERM was associated with ocular inflammation and one case of combined hamartoma of the retina and retinal pigment epithelium (RPE). In 13 cases, the ERM was especially white, thick, and opaque, with localized constriction and severe retinal distortion. Final best-corrected visual acuity (VA) was significantly better than preoperatively (20/50 vs. 20/112), (P = 0.0002). Mean improvement in VA was 4.25 lines and 17 patients gained two or more lines. This improvement was better in secondary than idiopathic ERM (6 lines vs. 3.3). During follow up, five cases of recurrence were observed (25%). In another case, postoperative persistent ocular hypertension required filtering surgery. CONCLUSION: The characteristics of ERM in young patients are quite different in many cases from those in adults in terms of thickness and adherence. Removal of ERM in young patients is feasible and safe although the membrane may focally adhere strongly to retinal vessels. VA usually improves significantly after surgery, but recurrences are more frequent than in adults. PMID- 11860974 TI - A cost-utility analysis of interventions for severe proliferative vitreoretinopathy. AB - PURPOSE: The purpose of this study was to report a reference case, patient preference-based, incremental, cost-utility analysis for treatments of retinal detachment associated with severe proliferative vitreoretinopathy (PVR). DESIGN: Computer-based economic model utilizing data from the Medicare health insurance system in the United States. METHODS: A cost-utility analysis compared vitreoretinal surgery using expanding gases and silicone oil therapy to the natural course of retinal detachment associated with severe PVR. The model applies long-term published visual data from the Silicone Study Group, time tradeoff utility analysis, decision analysis with Markov modeling, and discounting of costs and health benefits as per the Panel on Cost-Effectiveness in Health and Medicine. The major outcome measure was in year 2000 United States dollars per quality-adjusted life-year (dollars/QALY) gained. RESULTS: Vitreoretinal surgery for retinal detachment complicated by severe PVR, as compared with no treatment, resulted in a mean gain of 0.128-0.200 discounted (3% annual rate) quality-adjusted life-years per treated patient. Silicone oil (dollars/QALY gained of 40,252 dollars) was slightly more cost-effective than perfluoropropane (C(3)F(8)) gas (dollars/QALY gained of 46,926 dollars) in eyes with PVR without previous vitrectomy, whereas C(3)F(8) gas (dollars/QALY gained of 46,162 dollars) was more cost-effective than silicone oil (dollars/QALY gained of 62,383 dollars) with previous vitrectomy and PVR. Sensitivity analysis resulted in a dollars/QALY gained of 13,347 dollars when 10% of opposite eyes had a severe visual loss to 202,128 dollars when a discount rate of 10% was utilized and opposite eyes initially had good vision. CONCLUSIONS: The incremental expense of interventions for retinal detachment associated with PVR is cost-effective when compared with other widely accepted interventional therapies across diverse medical specialties. PMID- 11860975 TI - Pathologic features of vascular endothelial growth factor-induced retinopathy in the nonhuman primate. AB - PURPOSE: Vascular endothelial growth factor (VEGF) is a potent ischemia upregulated angiogenic protein that has been implicated in diabetic retinopathy. Intravitreal VEGF injections have not previously been shown to produce preretinal neovascularization. The purpose of this study was to further characterize the angiopathic changes that occur after intravitreal injections in a nonhuman primate and determine if preretinal neovascularization develops. DESIGN: Experimental animal study. METHODS: Vascular endothelial growth factor 165 was injected into the eyes of normal cynomolgus monkeys at regular intervals. As a control, normal eyes were injected with phosphate buffered saline. Color photography and fluorescein angiography were performed at regular intervals. The retinas were incubated for adenosine diphosphatase (ADPase) activity to visualize retinal vessels. The retinas were flat-embedded and areas of potential preretinal neovascularization were identified en bloc and serially sectioned. RESULTS: Areas of capillary nonperfusion and vessel dilation and tortuousity were seen by angiography. In serial sections, the nonperfused areas were found to be associated with endothelial cell hyperplasia in vessel lumens. Preretinal neovascularization originating only from superficial veins and venules was observed throughout peripheral retina, but was not seen in the posterior pole. Lacunae-like veins were subdivided by the process of intussusception and endothelial cell bridging. Arterioles demonstrated endothelial cell hyperplasia and microaneurysms. CONCLUSION: Intraocular injections of VEGF were sufficient to produce preretinal neovascularization in the nonhuman primate. Most vasculopathic structures were associated with endothelial cell hyperplasia. These results demonstrate that VEGF alone can produce many features of both nonproliferative and proliferative diabetic retinopathy including the previously undescribed development of preretinal neovascularization. This well-characterized VEGF induced primate model of retinal neovascularization may be useful as a means of testing new treatments for retinal neovascularization. PMID- 11860976 TI - Community screening for eye disease by laypersons: the Hoffberger program. AB - PURPOSE: To describe the results of a community-based eye screening program in Baltimore. DESIGN: Cross-sectional study. METHODS: This was a retrospective study of the results of screening both eyes of 5352 persons who presented at multiple community sites. The screening examination had eight risk factor questions, visual acuity measurement, and a screening field test and was carried out by technicians and lay volunteers. Screened persons (screenees) received a definitive eye examination at no out-of-pocket cost, transportation was offered, and inexpensive eyeglasses were provided if needed. The main outcome measures were the rate of appointment keeping and the eye diseases identified. Telephone interviews were used to assess reasons for missing appointments and satisfaction with visits. RESULTS: Screenees had a median age of 45 years, were 71% black, 59% female, and had estimated median annual family income of 24,000 dollars. Among 1331 screenees who scheduled a definitive examination appointment, 552 (41%) completed the visit. Data on definitive diagnosis was available in 480 out of 552 persons (87%). Reasons given for failing to come for definitive examination were: no appointment given (26%), forgot (20%), lack of transportation (9%), and lack of insurance coverage (6%). Of those who accepted a second visit date after defaulting, only 25% (41/167) appeared. Of 17 persons identified with glaucoma at screening, 4 had previously been diagnosed, but had ceased active care. CONCLUSION: After community screening for eye disease, efforts to provide definitive ophthalmic examination were only modestly effective. Failure of screenees to come for examination and loss to follow up were identified as serious problems. PMID- 11860977 TI - Polymerase chain reaction diagnosis in fungal keratitis caused by Alternaria alternata. AB - PURPOSE: To contribute toward assessing the effectiveness of polymerase chain reaction as a rapid method in diagnosis of torpid keratitis caused by opportunistic fungi. METHODS: Interventional case report. A 50-year-old man with a corneal abscess in the right eye treated for a period of 6 months with different combinations of broad-spectrum antibiotics and steroids was referred to our center. Corneal scraping was taken for microbiological study, including classic cultures and polymerase chain reaction. Amplified DNA was sequenced to identify the pathogen. RESULTS: Polymerase chain reaction amplification was negative for Acanthamoeba species and positive for fungi. The sequence analysis showed Alternaria alternata as the causal agent in 24 hours. Cultures confirmed the identification in 10 days. CONCLUSION: Polymerase chain reaction amplification with subsequent DNA-typing was revealed to be a useful method for detection of ocular pathogens such as A. alternata involved in cases of torpid keratitis, even in the presence of broad-spectrum antimicrobial therapy. PMID- 11860978 TI - Corneal edema with dislocated anterior polar cataract. AB - PURPOSE: To report an unusual manifestation of anterior polar cataract. METHODS: Observational case report. A 35-year-old woman presented with localized inferior corneal edema associated with a white nodular lesion on the iris. The anterior lens capsule was deficient centrally with scrolled edges and an underlying subcapsular opacity. The nodule was removed surgically and subjected for histopathologic examination. RESULTS: The histopathologic examination of the white nodular lesion showed parallel collagen fibers that were positive for collagen on the Masson trichrome stain. Correlating with clinical features, these findings were suggestive of dislocated anterior polar cataract. CONCLUSION: Anterior pyramidal cataract may dislocate spontaneously and present as an anterior chamber foreign body and may even lead to corneal endothelial cell loss. PMID- 11860979 TI - Reactivation of herpes simplex virus keratitis after initiating bimatoprost treatment for glaucoma. AB - PURPOSE: To report a case of herpes simplex virus reactivation after starting bimatoprost treatment for glaucoma. DESIGN: Interventional case report. METHODS: A 66-year-old woman had a herpes simplex keratouveitis reactivation that occurred within 1 month after starting bimatoprost. The herpes simplex had been inactive for more than 10 years. RESULTS: Bimatoprost and prednisolone acetate 0.12% were discontinued; oral acyclovir, ofloxacin, and betaxolol 0.25% were initiated. Two weeks later, prednisolone acetate 1% was added. The reactivation resolved, and 1 month later, the best corrected visual acuity improved to 20/40. CONCLUSION: Caution should be used in prescribing bimatoprost for patients with a history of herpes simplex virus keratitis. PMID- 11860980 TI - Cystoid macular edema associated with ocular hypotensive lipids. AB - PURPOSE: To report angiographically documented cystoid macula edema associated with the use of each of the three newly available ocular hypotensive lipids: unoprostone, travaprost, and bimatoprost. DESIGN: Observational case series. METHODS: Retrospective review of three patients in a clinical practice who had uncontrolled glaucoma on maximal tolerable therapy except for an ocular hypotensive lipids. All three patients also had previous cataract and filtration surgery, and all had an absent or open posterior lens capsule. The patients were informed of the potential risks of cystoid macula edema associated with the use of an ocular hypotensive lipids versus the risks of repeat filtration surgery. RESULTS: An ocular hypotensive lipids was started in the affected eye in each patient, and the patient was instructed to check visual acuity everyday and report back any change in vision occurred. CONCLUSION: Decreased vision of at least two lines caused by angiographically confirmed cystoid macula edema was noted in each of three patients started, respectively, on unoprostone, travaprost, and bimatoprost. The visual acuity returned to baseline, and the cystoid macula edema was angiographically resolved after discontinuation of the ocular hypotensive lipids and the initiation of a topical steroid and non steroidal anti-inflammatory eyedrops. Until a causal relationship between cystoid macula edema and ocular hypotensive lipids is proved or disproved, caution in their use in high-risk eyes would be prudent. PMID- 11860981 TI - Endoscopic photocoagulation of the ciliary body after repeated failure of trans scleral diode-laser cyclophotocoagulation. AB - PURPOSE: To report the successful control of intraocular pressure with endoscopic cyclophotocoagulation after repeated failure of trans-scleral diode-laser cyclophotocoagulation. DESIGN: Interventional case report. METHODS: A 3 1/2-year old child had intraocular pressure around 30 mm Hg in one eye despite full medical treatment and three procedures of trans-scleral diode laser photocoagulation. RESULTS: Endoscopic view of the ciliary body revealed many misplaced laser burns in the pars plana region. Using direct endoscopic visualization of the ciliary body, precise, confluent burns were applied to the ciliary body. Six months after the procedure, intraocular pressure has remained under 20 mm Hg. CONCLUSION: Endoscopic cyclophotocoagulation can effectively lower intraocular pressure after recurrent external diode-laser photocoagulation has failed to do so. The direct visualization during the endoscopic procedure is particularly advantageous in cases with atypical ciliary body morphology, such as in pediatric glaucoma. PMID- 11860982 TI - Recurring transitory blindness caused by primary marginal pigment epithelial iris cysts. AB - PURPOSE: To report four cases of primary pupillary pigment epithelial iris cysts, all members of one family, in which two of the patients had recurring transitory visual impairment. METHODS: Observational case series. History was taken, the patients were examined with slit-lamp and ultrasound biomicroscopy, and surgically removed cyst tissue was examined with transmission electron microscopy. RESULTS: Pupillary pigment epithelial cysts of the iris generally show an autosomal dominant heredity pattern, with occasional lack of penetrance. In two of our cases, the size and location of the cysts caused visual symptoms, necessitating surgical removal. The cyst wall consists entirely of pigment epithelial cells. CONCLUSION: The origin of pupillary pigment epithelial cysts is unclear, and a hereditary background is very likely. Their clinical significance is in their similarity to pigmented tumors of the iris. They may also be indicative of coexisting systemic disease. In symptomatic cases, treatment is indicated. PMID- 11860983 TI - Foveal hypoplasia in oculocutaneous albinism demonstrated by optical coherence tomography. AB - PURPOSE: To document, in vivo, the foveal morphology and thickness in a patient with oculocutaneous albinism. METHODS: Observational case report. In a 10-year old female with oculocutaneous albinism, multiple cross-sectional scans of the fovea were performed using optical coherence tomography. RESULTS: Optical coherence tomography scans were unable to detect the foveal pit. A widespread thickening of the retina occurred throughout the entire fovea with no difference from the surrounding macula. The foveal thickness was greater than 300 microm in the eyes of this patient with oculocutaneous albinism, compared with 150 microm in the normal eye. The inner retina had a highly reflective signal on optical coherence tomography. CONCLUSION: Optical coherence tomography demonstrated in the anatomical location of the fovea a highly reflective inner retinal signal, possibly consistent with multiple layers of ganglion cells, and it confirmed foveal hypoplasia in a patient with oculocutaneous albinism. PMID- 11860984 TI - Autosomal recessive cerebellar ataxia with bull's-eye macular dystrophy. AB - PURPOSE: In 1980, we published in the American Journal of Ophthalmology two siblings with hereditary ataxia and atrophic maculopathy. The report is cited in the literature as autosomal dominant cerebellar ataxia with retinal degeneration. The purpose of the present study is to document the progression of the neurodegenerative disorder and to review the diagnosis. DESIGN: Observational case report. METHODS: Twenty years after the original publication, the 52-year old male patient had new ocular and neurologic examinations, fluorescein angiography, molecular genetic analysis, and biochemical testing. RESULTS: Fluorescein angiography showed marked progression of the macular dystrophy to a bull's-eye configuration. Genetic analysis of the patient did not show CAG trinucleotide repeat expansion in the various spinocerebellar ataxia genes. This excludes the diagnosis of autosomal dominant cerebellar ataxia with macular degeneration (ADCA type II) with mutation of the spinocerebellar ataxia 7 gene. Major causes of autosomal recessive cerebellar ataxia with retinal degeneration, including Friedreich ataxia and congenital disorders of glycosylation, were also excluded. CONCLUSION: The two patients, previously published in the American Journal of Ophthalmology by Eerola and coworkers, did not suffer from presently recognized disorders with cerebellar ataxia and retinal degeneration. The Eerola syndrome probably represents a separate neurodegenerative entity characterized by autosomal recessive cerebellar ataxia and progressive macular dystrophy with a bull's-eye pattern. PMID- 11860985 TI - Scanning laser ophthalmoscope findings in acute macular neuroretinopathy. AB - PURPOSE: The typical reddish-brown retinal lesions of acute macular neuroretinopathy are difficult to detect. We demonstrate a role for scanning laser ophthalmoscopy in the diagnosis of acute macular neuroretinopathy. METHODS: Observational case report. We used scanning laser ophthalmoscopy in a 23-year-old woman with acute macular neuroretinopathy. RESULTS: In contrast to biomicroscopy, color fundus photography, and red-free fundus photography, scanning laser ophthalmoscopy precisely disclosed the size of the lesions within the macula. Microperimetry showed absolute scotomata corresponding to the macular lesions. CONCLUSION: Scanning laser ophthalmoscopy enhances the visibility of the retinal lesions in acute macular neuroretinopathy. The lesion size can be determined more precisely, and follow-up of patients is more accurate compared with conventional techniques. PMID- 11860986 TI - Hemiretinal vein occlusion associated with membranous glomerulonephritis. AB - PURPOSE: To report a patient in whom the finding of hemiretinal vein occlusion led to the diagnosis of membranous glomerulonephritis. DESIGN: Interventional case report. METHODS: A 44-year-old tennis instructor presented with a 1-week history of blurred vision in the left eye. Examination of the left eye demonstrated a best-corrected visual acuity of 20/40 and an inferior hemiretinal vein occlusion. RESULTS: Blood pressure was normal, and the patient was referred for a medical examination, which revealed membranous glomerulonephritis. The patient was treated with oral prednisone and cyclosporine. Four months after presentation, the left eye demonstrated resolution of the vascular abnormalities and had a best-corrected visual acuity of 20/20. CONCLUSION: Retinal vein occlusion may be associated with membranous glomerulonephritis. Treatment of the systemic disease may be associated with regression of the retinal vascular abnormalities. PMID- 11860987 TI - Multifocal electroretinography response after laser photocoagulation of a subretinal nematode. AB - PURPOSE: To describe multifocal electroretinography findings before and after laser photocoagulation of a subretinal nematode in diffuse unilateral subacute neuroretinitis. METHOD: Observational case report. A 45-year-old woman with left eye inflammation, subretinal tracts superior and temporal to the fovea, and a subretinal coiled mobile parasite was treated with laser photocoagulation to destroy the nematode. Multifocal electroretinography was performed before and after laser photocoagulation. RESULTS: In the left eye, multifocal electroretinography before treatment showed decreased foveal response density and increased parafoveal and perifoveal waveform amplitudes. Two months after laser photocoagulation, multifocal electroretinography showed full recovery of normal findings and the visual acuity remained 20/20. CONCLUSION: Multifocal electroretinography appears to be useful in evaluating the retinal findings after photocoagulation of a parasite associated with diffuse unilateral subacute neuroretinitis. PMID- 11860988 TI - Optical coherence tomography of choroidal osteoma. AB - PURPOSE: To examine the cross-sectional structure of a choroidal osteoma using optical coherence tomography. METHODS: Observational case report. A choroidal osteoma in the right eye of a 22-year-old woman was studied with fluorescein and indocyanine green fundus angiograms and optical coherence tomography. RESULTS: The optical coherence tomography showed that a creamy-white lesion in the tumor had an increased reflectivity with a cavernous structure, whereas an orange lesion showed less reflectivity beneath the apparently intact retinal pigment epithelium. The area of less-intense whiteness in the creamy-white lesion showed higher reflectivity at the level of the retinal pigment epithelium, which precluded our viewing the deeper structure of the tumor. CONCLUSION: Optical coherence tomography can demonstrate the cavernous structure of a choroidal osteoma at a selective stage of a tumor's evolution, namely, when the retinal pigment epithelium is atrophied and the ossification is premature. PMID- 11860989 TI - Two new forceps for use during and after dacryocystorhinostomy. AB - PURPOSE: To report two new forceps for use during and after external dacryocystorhinostomy (E-DCR). Description of surgical instruments is provided. METHODS: Interventional case series. The E-DCR forceps were modified from the usual ophthalmic forceps, and they were designed for grasping the posterior flaps in the narrow and deep surgical region. The washout forceps under the transnasal endoscopic (TNE) examination consists of the usual otolaryngeal forceps with an injector tip at the apex to wash out discharge around the ostium in the nasal cavity. RESULTS: In consecutive 28 E-DCR procedures cases (25 patients), E-DCR was successfully performed using the E-DCR forceps. This forceps was useful to suture the posterior flaps in all cases. The washout forceps was available to remove the congealed discharge that could not be aspirated. CONCLUSION: These two forceps proved to be useful during and after the E-DCR procedure. PMID- 11860990 TI - Spontaneous retrobulbar hemorrhage in type IV Ehlers-Danlos syndrome. AB - PURPOSE: To describe a case of spontaneous retrobulbar hemorrhage in type IV Ehlers-Danlos syndrome, a disorder characterized by vascular fragility and associated severe bleeding problems. METHODS: Observational case report. Spontaneous unilateral retrobulbar hemorrhage in a 22-year-old man with type IV Ehlers-Danlos syndrome was documented by clinical and radiographic examination. RESULTS: The retrobulbar hemorrhage was self-limited and resolved without visual sequelae. CONCLUSION: Spontaneous retrobulbar hemorrhage should be recognized as part of the spectrum of type IV Ehlers-Danlos syndrome, and clinicians must be alert to diagnosis and treatment of this vision-threatening problem. PMID- 11860991 TI - Lymphomatous meningitis of the Burkitt type presenting with multiple cranial neuropathies. AB - PURPOSE: To describe diplopia as the initial manifestation of the Burkitt lymphoma. DESIGN: Observational case report. METHODS: Retrospective chart review. RESULTS: A 53-year-old human immunodeficiency virus (HIV)-positive man presented with a severe headache associated with binocular diplopia. Flow cytometric analysis of the cerebrospinal fluid demonstrated a monoclonal B-lymphoid cell population consistent with the Burkitt lymphoma. CONCLUSION: Ophthalmologists should be aware that diplopia may be the presenting manifestation of Burkitt lymphoma in an immunocompromised patient. PMID- 11860992 TI - Accommodation and convergence palsy caused by lesions in the bilateral rostral superior colliculus. AB - PURPOSE: To report a patient who developed accommodation and convergence palsy caused by lesions in the bilateral rostral superior colliculus. DESIGN: Observational case report. METHODS: A 30-year-old right-handed man experienced sudden onset of diplopia and blurred vision at near vision. RESULTS: The patient showed accommodation and convergence palsy. Magnetic resonance imaging revealed lesions located in the bilateral rostral superior colliculus. CONCLUSION: These findings suggest that the rostral superior colliculus is involved in the control of accommodation and vergence eye movements. PMID- 11860993 TI - Perimetric sensitivity and retinal thickness in eyes with macular edema resulting from branch retinal vein occlusion. PMID- 11860994 TI - Silicone oil-associated optic nerve degeneration. PMID- 11861011 TI - Synthesis of alpha-Manp-(1-->2)-alpha-Manp-(1-->3)-alpha-Manp-(1-->3)-Manp, the tetrasaccharide repeating unit of Escherichia coli O9a, and alpha-Manp-(1-->2) alpha-Manp-(1-->2)-alpha-Manp-(1-->3)-alpha-Manp-(1-->3)-Manp, the pentasaccharide repeating unit of E. coli O9 and Klebsiella O3. AB - The tetrasaccharide repeating unit of Escherichia coli O9a, alpha-D-Manp-(1-->2) alpha-D-Manp-(1-->3)-alpha-D-Manp-(1-->3)-D-Manp, and the pentasaccharide repeating unit of E. coli O9 and Klebsiella O3, alpha-D-Manp-(1-->2)-alpha-D-Manp (1-->2)-alpha-D-Manp-(1-->3)-alpha-D-Manp-(1-->3)-D-Manp, were synthesized as their methyl glycosides. Thus, selective 3-O-allylation of p-methoxyphenyl alpha D-mannopyranoside via a dibutyltin intermediate gave p-methoxyphenyl 3-O-allyl alpha-D-mannopyranoside (2) in good yield. Benzoylation (-->3), then removal of 1 O-methoxyphenyl (right arrow4), and subsequent trichloroacetimidation afforded the 3-O-allyl-2,4,6-tri-O-benzoyl-alpha-D-mannopyranosyl trichloroacetimidate (5). Condensation of 5 with methyl 4,6-O-benzylidene-alpha-D-mannopyranoside (6) selectively afforded the (1-->3)-linked disaccharide 7. Benzoylation of 7, debenzylidenation, benzoylation, and deallylation gave methyl 2,4,6-tri-O-benzoyl alpha-D-mannopyranosyl-(1-->3)-2,4,6-tri-O-benzoyl-alpha-D-mannopyranoside (11) as the disaccharide acceptor. Coupling of 11 with (1-->2)-linked mannose disaccharide donor 17 or trisaccharide donor 21, followed by deacylation, furnished the target tetrasaccharide and pentasaccharide, respectively. PMID- 11861012 TI - Synthesis of an xylosylated rhamnose pentasaccharide, the repeating unit of the O chain polysaccharide of the lipopolysaccharide of Xanthomonas campestris pv. begoniae GSPB 525. AB - A xylosylated rhamnose pentasaccharide, alpha-L-Rhap-(1-->3)-[beta-L-Xylp-(1-->2) ]-alpha-L-Rhap-(1-->3)-[beta-L-Xylp-(1-->4)]-L-Rhap, the repeating unit of the O chain polysaccharide (OPS) of the lipopolysaccharides of Xanthomonas campestris pv. begoniae GSPB 525 was synthesized by a highly regio- and stereoselective way. Thus coupling of 1,2-O-ethylidene-beta-L-rhamnopyranose (1) with 2,3,4-tri-O benzoyl-alpha-L-rhamnopyranosyl trichloroacetimidate (2) to give (1-->3)-linked disaccharide (3), subsequent benzoylation, deethylidenation, acetylation, 1-O deacetylation, and trichloroacetimidation afforded the disaccharide donor 11. Condensation of 11 with 1 yielded 2,3,4-tri-O-benzoyl-alpha-L-rhamnopyranosyl-(1- >3)-2-O-acetyl-4-O-benzoyl-alpha-L-rhamnopyranosyl-(1-->3)-1,2-O-ethylidene-beta L-rhamnopyranose (12), and selective deacetylation of 12 yielded the trisaccharide diol acceptor 15. Coupling of 15 with 2,3,4-tri-O-benzoyl-alpha-L xylopyranosyl trichloroacetimidate (16), followed by deprotection, gave the target pentasaccharide 19. PMID- 11861013 TI - Efficient synthesis of 2-deoxy-L-erythro-pentose (2-deoxy-L-ribose) from L arabinose. AB - An efficient and practical route for the large-scale synthesis of 2-deoxy-L erythro-pentose (2-deoxy-L-ribose) starting from L-arabinose was developed using Barton-type free-radical deoxygenation reaction as a key step. The radical precursor, a phenoxythiocarbonyl ester, was prepared in situ, and the most efficient deoxygenation was achieved by slow addition of tributyltin hydride to the reaction mixture. PMID- 11861014 TI - Flavonoid glycosides from Salvia moorcroftiana wall. AB - Phytochemical analysis of the whole plant of Salvia moorcroftiana Wall. (Lamiaceae) resulted in the isolation of two new flavonoid glycosides, together with three known compounds. The structures of the new compounds were established as genkwanin 4'-O-alpha-L-arabinopyranosyl-(1-->6)-beta-D-galactopyranoside (1) and genkwanin 4'-O-[alpha-L-rhamnopyranosyl-(1-->2)-[alpha-L-rhamnopyranosyl-(1- >6)]-beta-D-galactopyranoside] (2). The structures of all compounds were elucidated by spectroscopic methods, including 2D NMR techniques. PMID- 11861015 TI - Structural analysis of lipopolysaccharide oligosaccharide epitopes expressed by non-typeable Haemophilus influenzae strain 176. AB - The structure of the lipopolysaccharide (LPS) from non-typeable Haemophilus influenzae strain 176 has been investigated. Electrospray ionization-mass spectrometry (ESIMS) on O-deacylated LPS (LPS-OH) and core oligosaccharide (OS) samples obtained after mild-acid hydrolysis of LPS provided information on the composition and relative abundance of the glycoforms. ESIMS tandem-mass spectrometry on LPS-OH confirmed the presence of minor sialylated and disialylated glycoforms. Oligosaccharide samples were studied in detail using high-field NMR techniques. It was found that the LPS contains the common inner core element of H. influenzae, L-alpha-D-Hepp-(1-->2)-[PEtn-->6]-L-alpha-D-Hepp (1-->3)-[beta-D-Glcp-(1-->4)]-L-alpha-D-Hepp-(1-->5)-[PPEtn-->4]-alpha-Kdop-(2- >6)-Lipid A having glycosyl substitution at the O-3 position of the terminal heptose as recently observed for non-typeable H. influenzae strain 486 [Mansson, M.; Bauer, S. H. J.; Hood, D. W.; Richards, J. C.; Moxon, E. R.; Schweda, E. K. H., Eur. J. Biochem. 2001, 268, 2148--2159]. The following LPS structures were identified as the major glycoforms, the most significant being indicated with an asterisk (*) (glycoforms are partly substituted with Gly at the terminal Hep): PMID- 11861016 TI - Effect of roasting on degradation and structural features of polysaccharides in Arabica coffee beans. AB - The degree and nature of polysaccharide degradation at different roasting levels was determined for three Arabica (Coffea arabica) bean varieties. Between 12 and 40% of the bean polysaccharides were degraded depending on the roasting conditions. The thermal stability of the arabinogalactans, (galacto)mannans and cellulose was markedly different. The arabinogalactans and mannans were degraded up to 60 and 36%, respectively, after a dark roast, while cellulose showed negligible evidence of degradation. Roasting led to increased solubility of both the arabinogalactans and (galacto)mannans from the bean but the structural modifications, which accompanied this change in solubility, were different for each polysaccharide. Despite the moderate degradation of the (galacto)mannans, those remaining in the bean after roasting showed no evidence of change to their molecular weight even after a dark roast. In contrast, arabinogalactans were depolymerised after a light roast both by fission of the galactan backbone and loss of arabinose from the sidechains. The recently discovered covalent link between the coffee bean arabinogalactans and protein survived roasting. The glucuronic acid component of the AG was degraded markedly after a dark roast, but approximately 30% of the original content remained as part of the AG polymer. The results show that polysaccharide degradation during roasting is more marked than previously documented, and points to roasting induced changes to the polysaccharides as major factors in the changing physicochemical profile of the coffee bean during processing. PMID- 11861017 TI - Structural studies on kappa-carrageenan derived oligosaccharides. AB - Oligosaccharides were prepared through mild hydrochloric acid hydrolysis of kappa carrageenan from Kappaphycus striatum carrageenan. Three oligosaccharides were purified by strong-anion exchange high-performance chromatography. Their structure was elucidated using mass spectral and NMR data. Negative-ion electrospray ionization (ESI) mass spectra at different fragmentor voltages provided the molecular weight of the compounds and unraveled the fragmentation pattern of the kappa-carrageenan oligosaccharides. 2D NMR techniques, including 1H-(1)H COSY, 1H-(1)H TOCSY and 13C-(1)H HMQC, were performed to determine the structure of a trisulfated pentasaccharide. 1D NMR and ESIMS were used to determine the structures of a kappa-carrageenan-derived pentasaccharide, heptasaccharide, and an undecasaccharide. All the oligosaccharides characterized have a 4-O-sulfo-D-galactopyranose residue at both the reducing and nonreducing ends. PMID- 11861018 TI - Effect of molecular structure on thermodynamic properties of carbohydrates. A calorimetric study of aqueous di- and oligosaccharides at subzero temperatures. AB - For aqueous solutions of di- and oligosaccharides thermodynamic properties have been investigated at subzero temperatures using differential scanning calorimetry. The amount of unfrozen water observed is found to increase linearly with the glass transition temperatures of anhydrous carbohydrates. Furthermore, the amount of unfrozen water shows a linear relationship with known solution properties of aqueous carbohydrates, such as partial molar compressibility and heat of solution. The different effectiveness among various di- and oligosaccharides to avoid ice formation is associated with the combination of constitutive monosaccharides and attendant molecular structure features including the position and type of the glycosidic linkage between the constituent units. More unfrozen water is induced in the presence of a carbohydrate having a poorer compatibility with the three-dimensional hydrogen-bond network of water. A series of these results obtained imply that there is a common key of carbohydrate stereochemistry governing several different thermodynamic amounts of a given system involving carbohydrates. In this context, a modified stereospecific hydration model can be used to interpret the present results in terms of stereochemical effects of carbohydrates. PMID- 11861019 TI - Preparative route to N-glycolylneuraminic acid phenyl 2-thioglycoside donor and synthesis of Neu5Gc-alpha-(2-->3')-lactosamine 3-aminopropyl glycoside. AB - The spacer-armed trisaccharide, Neu5Gc-alpha-(2-->3')-lactosamine 3-aminopropyl glycoside, was synthesized by regio- and stereoselective sialylation of the suitably protected triol acceptor, 3-trifluoroacetamidopropyl 2-acetamido-3,6-di O-benzyl-2-deoxy-4-O-(6-O-benzyl-beta-D-galactopyranosyl)-beta-D-glucopyranoside, with the donor methyl [phenyl 5-acetoxyacetamido-4,7,8,9-tetra-O-acetyl-3,5 dideoxy-2-thio-D-glycero-alpha,beta-D-galacto-2-nonulopyranosid]onate. The donor was obtained, in turn, from methyl [phenyl 5-acetamido-4,7,8,9-tetra-O-acetyl-3,5 dideoxy-2-thio-D-glycero-alpha,beta-D-galacto-2-nonulopyranosid]onate by N-tert butoxycarbonylation of the acetamido group followed by total N- and O deacetylation, per-O-acetylation, subsequent Boc group removal, and N acetoxyacetylation. PMID- 11861020 TI - Stereospecific synthesis of (+)-oxybiotin from D-xylose. AB - A new 14-step synthesis of (+)-oxybiotin, an oxygen analogue of (+)-biotin, was achieved starting from D-xylose by use of selected 2,5-anhydro sugar derivatives as key intermediates. PMID- 11861021 TI - O-Specific chain structure from the lipopolysaccharide fraction of Pseudomonas reactans: a pathogen of the cultivated mushrooms. AB - An O-specific polysaccharide containing 2-acetamidino-2-deoxy-beta-D glucopyranose (Glcp2Am), 2,4-diacetamido-2,4,6-trideoxy-beta-D-glucopyranose (QuipNAc4NAc, bacillosamine) and 2,4-di-(N-acetyl-L-alanylamino)-2,4,6-trideoxy beta-D-glucopyranose (QuipNAlaAc4NAlaAc) was isolated from the phenol-soluble lipopolysaccharide fraction of the mushroom-associated bacterium Pseudomonas reactans. The structure, determined by means of chemical analysis and 1D and 2D NMR spectroscopy, showed a linear trisaccharide-repeating unit, as shown below:- >3)-beta-D-QuipNAlaAc4NAlaAc-(1-->3)-alpha-D-Glcp2Am-(1-->3)-alpha-D QuipNAc4NAc(1-->To our knowledge, this is the first complete O-chain structure reported for the lipopolysaccharide of a mushroom-associated bacterium. PMID- 11861023 TI - Spotlight on gender. PMID- 11861024 TI - Risk factors for coronary heart disease: implications of gender. AB - It has been recognized over the past years that women form a distinct subpopulation within patients with coronary heart disease. This phenomenon should be acknowledged in the management and in the assessment of coronary heart disease. Over the past years remarkable progress has been made concerning our knowledge of cardiovascular risk factors related to gender. For instance, diabetes, high density lipoproteins and triglycerides levels have been found to have a greater impact on coronary heart disease risk in women compared to men. On the other hand, evidence showing that lipoprotein (a) is a cardiovascular risk factor seems to be stronger in men than in women. For optimal treatment and prevention of coronary heart disease it is necessary to acknowledge that it is not self-evident that women and men show similar responses to risk factors or to treatment. This review article addresses the role of cardiovascular risk factors focusing on the differential impact they might have on men and women. PMID- 11861025 TI - Hormones, genetic factors, and gender differences in cardiovascular disease. PMID- 11861026 TI - Clinical characteristics of coronary heart disease in women: emphasis on gender differences. PMID- 11861027 TI - Smoking and gender. AB - Smoking is a major cause of coronary heart disease for both men and women and a positive correlation between tobacco use and cerebrovascular disease has been also described. In addition, cigarette smoking is the most powerful risk factor predisposing to atherosclerotic peripheral artery disease. More recently, passive smoking has been also shown to represent an important risk factor for coronary artery disease. Moreover, the incidence of coronary artery and cerebrovascular diseases in ex-smokers consistently decreases after cessation, further underlying the relevance of smoking as a risk factor for these pathological conditions. The effects of cigarette smoking on atherosclerosis initiation and progression as well on its complications are mostly responsible for the enhanced cardio- and cerebrovascular risk observed in smoking compared to non-smoking subjects. Since hormonal status may also play a role in the development and stability of the atherosclerotic plaque, smoking habits could influence the clinical complications of atheroclerosis in a gender dependent manner. Up to now, however, few studies have investigated the relative importance of smoking as a risk factor for fatal and non-fatal diseases in the two sexes within the same study population. On the basis of available clinical data, this review will discuss the risk of fatal and non-fatal diseases among smoking men and women with special emphasis on cardiovascular and cerebrovascular disease which also represents the most common cause of death among smokers. A description of the mechanisms involved in the tobacco-induced atherosclerotic damage will be also given in order to underline possible gender-related differences. PMID- 11861028 TI - Estrogen and homocysteine. AB - Cardiovascular diseases are the major causes of illness and death in women. Premenopausal women are relatively protected from coronary artery disease and atherosclerosis as compared to postmenopausal women, and this protection is attributed to the effects of the female sex hormone (estrogen). The vasculature, like the reproductive tissues, bone, liver, and brain, is now recognized as an important site of estrogen's action. Although estrogen's beneficial effects on the cardiovascular system are well described in many studies, the molecular basis of estrogen protective mechanisms are still quite vague. Both genomic mechanisms, mediated primarily through estrogen receptor alpha (ER alpha) and estrogen receptor beta (ER beta), and non-genomic mechanisms, through nitric oxide (NO), of estrogen action are controversial and do not entirely explain the effects of estrogen on vascular preservation during conditions of oxidative stress. Until recently, the atheroprotective effects of estrogen were attributed principally to its effects on serum lipid concentrations and cholesterol levels. However, two recent reports that estrogen therapy has no effect on the progression of coronary atherosclerosis in women with established disease, despite the favorable changes in LDL and cholesterol levels, leads to questions about the lipid/cholesterol mechanism of estrogen-mediated effects on atherosclerosis. Alternatively, the high level of homocysteine, found to correlate with accelerated cardiovascular disease and identified as an independent risk factor for atherosclerosis, was recently described to be diminished by estrogen. Protection against disturbed sulfhydryl metabolism and higher homocysteine level could be the missing link in understanding how exactly estrogen affects vascular cells metabolism and responses to oxidative stress. This review focuses on estrogen/homocysteine interactions and their relevance to the cardiovascular system. PMID- 11861029 TI - Estradiol prevents homocysteine-induced endothelial injury in male rats. AB - OBJECTIVE: We investigated whether estradiol may prevent accelerated atherosclerosis due to hyperhomocysteinemia by enhancing the antioxidant system. METHODS: Male Wistar rats were treated with placebo (P) or 1 mg (1E2) and 2 mg (2E2) 17 beta estradiol. Half of the animals (n=6) from each group received homocysteine (Hcy, 100 mg/kg/day) administered in the drinking water for 60 days (P/Hcy, 1E2/Hcy and 2E2/Hcy). Glutathione (GSH) content and glucose-6-phosphate dehydrogenase (G6PDH) activity were determined in myocardial tissues, as well as the serum Hcy concentrations and blood levels of hydrogen peroxide (H(2)O(2)). The relaxation response of aortic ring segments to acetylcholine (ACh) was used for the assessment of endothelial function, and hematoxylin-eosin stained thin sections of rat aorta were used for detection of the histological changes (namely endothelial damage and wall thickening). RESULTS: Depression of relaxation to ACh occurred in P/Hcy compared to P (15.7 +/- 4% vs. 96.3 +/- 7%, P<0.0001), but estrogen significantly restored endothelium dependent relaxation in hyperhomocysteinemic rats (86.8 +/- 9.3%, P<0.001). Histological examination revealed aortic endothelial denudation in P/Hcy while the endothelial structures of the aorta from the 1E2/Hcy and 2E2/Hcy appeared normal. Significant reductions in GSH and G6PDH levels were detected in P/Hcy (1.5 +/- 0.01 micromol/g and 3.21 +/- 1.2 U/mg, respectively) compared to 1E2/Hcy (2.5 +/- 0.3 micromol/g and 12.81 +/- 1.5 U/mg, P<0.001) and 2E2/Hcy (3.11 +/- 1.1 micromol/g and 15.66 +/- 4 U/mg, P<0.001). In addition, blood H(2)O(2) level in 1E2/Hcy and 2E2/Hcy remained low while it was raised significantly in P/Hcy compared to P (P<0.001). CONCLUSIONS: These data suggest that the observed reduction of GSH levels and suppression of G6PDH activity induced by Hcy coupled, with endothelial ultrastructural changes and impaired function, all reversed by estradiol, may have relevance to the mechanisms of atherogenesis and the beneficial effects of estrogen replacement therapy. PMID- 11861030 TI - Endothelial function, vascular reactivity and gender differences in the cardiovascular system. PMID- 11861031 TI - Estrogen replacement therapy, atherosclerosis, and vascular function. AB - There is strong evidence from both human and nonhuman primate studies supporting the conclusion that estrogen deficiency increases the progression of atherosclerosis. More controversial is the conclusion that postmenopausal estrogen replacement inhibits the progression of atherosclerosis. Estrogen treatment of older women (>65 years) with pre-existing coronary artery atherosclerosis had no beneficial effects. In contrast, estrogen treatment of younger postmenopausal women or monkeys in the early stages of atherosclerosis progression has marked beneficial effects. Whether progestogens attenuate the cardiovascular benefits of estrogen replacement therapy has been controversial for more than a decade. Current evidence from studies of both monkeys and women suggest little or no attenuation of estrogen benefits for coronary artery atherosclerosis. Lack of compliance with estrogen replacement therapy, usually because of fear of breast cancer, remains a major problem. Future regimens may overcome that fear by the co-administration of a breast cancer preventive agent (i.e., selective estrogen receptor modulators, phytoestrogens) with low dose estrogen. PMID- 11861032 TI - Overview of gender aspects of cardiac syndrome X. AB - Cardiac syndrome X, a condition defined by the presence of angina-like chest pain, a positive response to stress testing and normal coronary arteriograms, has been shown to occur in approximately 20--30% of angina patients undergoing coronary arteriography. The prevalence of syndrome X is significantly higher in women compared to men. In the majority of patients with chest pain and normal coronary arteriograms, symptoms are likely to be non-cardiac in origin. However, myocardial ischaemia may be the pathogenic mechanism in a proportion of syndrome X patients. Indeed, the clinical characteristics, the ischaemic electrocardiographic findings and the presence of myocardial perfusion defects during stress testing are similar in syndrome X and coronary artery disease patients. Moreover, coronary sinus oxygen saturation abnormalities and pH changes, as well as myocardial lactate production and alterations of cardiac high energy phosphate are seen during stress testing in patients with syndrome X and appear to endorse an ischaemic origin of symptoms in at least a proportion of these individuals. Patients with chest pain and normal coronary arteries have abnormal vasodilatory coronary blood flow responses and an increased sensitivity of the coronary microcirculation to vasoconstrictor stimuli (microvascular angina). Microvascular endothelial dysfunction appears to be responsible for these coronary microcirculation abnormalities. Given the high prevalence of peri- and post-menopausal women in cardiac syndrome X, it has been hypothesized that oestrogen deficiency may play a major role in the pathogenesis of this condition. Oestrogen vasoactive properties involve endothelium-dependent effects and, in postmenopausal women, forearm vasodilatation induced by acetylcholine is potentiated by the acute local administration of intravenous oestradiol. This suggests that endothelium-dependent responses in the peripheral circulation may be modulated by steroid hormones. Impairment of endothelial function in post menopausal women with syndrome X has been reported by various groups and it could be hypothesized that oestrogen deficiency may contribute to the development of microvascular angina through endothelial dysfunction and that exogenous oestrogen administration may have a beneficial effect in syndrome X patients. This article reviews current knowledge regarding the role of oestrogen deficiency in the pathogenesis of syndrome X and the potential therapeutic role of oestrogen replacement therapy in women with chest pain and normal coronary arteriograms PMID- 11861033 TI - Isoproterenol amplifies 17 beta-estradiol-mediated vasorelaxation: role of endothelium/nitric oxide and cyclic AMP. AB - OBJECTIVES: Estrogen exerts cardiac protection via multiple cellular mechanisms. Estrogen modifies vasodilatation induced by certain relaxants such as beta adrenoceptor agonists. However, little is known whether low concentrations of beta-adrenoceptor agonists would reciprocally influence the acute relaxant response to estrogen. The present study was designed to investigate the synergistic interaction between isoproterenol and 17 beta-estradiol, and the role of endothelium and cyclic AMP-dependent pathway in this interaction. METHODS: Changes in vessel tone of the isolated rat mesenteric artery rings were measured using a force-displacement Grass transducer. RESULTS: In 9,11-dideoxy-11 alpha, 9 alpha-epoxy-methanoprostaglandin F(2 alpha)-preconstricted endothelium-intact rings, 17 beta-estradiol induced relaxations with pD(2) of 5.06 +/- 0.06. Pretreatment of endothelium-intact rings with isoproterenol (1-3 x 10(-9) M, 1 h incubation time) significantly enhanced 17 beta-estradiol-induced relaxation. This effect was inhibited by Rp-cGMPS triethylamine (3 x 10(-6) M), and abolished in the presence of 3 x 10(-5) M N(G)-nitro-L-arginine methyl ester or in endothelium-denuded rings. The effect of isoproterenol was antagonized by propranolol (3 x 10(-6) M), ICI 118,551 (3 x 10(-6) M), but not by atenolol (10( 5) M). Rp-cAMPS triethylamine (3 x 10(-6) M) abolished the effect of isoproterenol. Besides, exposure to 3 x 10(-9) M forskolin for 1 h also potentiated the relaxant response to 17 beta-estradiol. CONCLUSION: In endothelium-intact rat mesenteric arteries pretreatment with low concentrations of isoproterenol enhanced the acute relaxant response to 17 beta-estradiol. This enhancement was dependent on the presence of endothelium and abolished by L-NAME via a beta(2)-adrenoceptor-mediated cyclic AMP-dependent mechanism. PMID- 11861034 TI - Estrogen replacement suppresses function of thrombin stimulated platelets by inhibiting Ca(2+) influx and raising cyclic adenosine monophosphate. AB - OBJECTIVE: Estrogen replacement therapy (ERT) in postmenopausal women reduces the risk of cardiovascular diseases. Beneficial changes in lipid profiles account for only one part, thereby raising the question of other estrogen induced benefit that may be lost at menopause. The purpose of this study was to determine the effects of estrogen replacement therapy (ERT) on platelet function of postmenopausal women. METHODS: The effect of 4 weeks ERT (conjugated estrogens 0.625 mg/day) on platelet function was evaluated ex vivo in 18 postmenopausal women (mean age 53 +/- 5 years, after menopause 3.8 +/- 1.9 years). RESULTS: After ERT, (1) plasma concentrations of estrone and estradiol significantly increased (estrone: 16 +/- 7-211+/- 80 pg/ml, estradiol: 14+/- 3-125 +/- 49 pg/ml, P<0.05) and LDL-cholesterol decreased (129 +/- 23-94 +/- 25 mg/dl, P<0.05). Plasma 6-keto-PG F(1) alpha significantly increased (7.2 +/- 3.4-13.3 +/ 6.7 pg/dl, P<0.05). (2) platelet aggregation and positive staining for P selectin in thrombin- (0.1 and 1.0 U/ml) stimulated platelets were inhibited (Th 0.1 U/ml: 4.0 +/- 0.9-2.4 +/- 1.0/control, P<0.05), but positive staining for GP IIb-IIIa complex did not alter significantly. (3) Ca(2+) influx induced by thrombin decreased (Th 0.3 U/ml: 345 +/- 29-298 +/- 24 nmol/l, P<0.05). The baseline [Ca(2+)](i), the release of Ca(2+) from internal stores induced by thrombin and the size of internal Ca(2+) stores did not alter. (4) platelet c-AMP increased (Th 0.3 U/ml: 66.4 +/- 9.4-82.6 +/- 13.0 fmol/l, P<0.05), but platelet nitrite/nitrate (NO(x)) or c-GMP did not alter significantly. CONCLUSIONS: These results suggest that modulation of platelet function by decreasing Ca influx and increased production of c-AMP may account in part for the cardiovascular benefit of ERT. PMID- 11861035 TI - The effect of 17 beta-estradiol on MCP-1 serum levels in postmenopausal women. AB - OBJECTIVE: Monocyte chemoattractant protein-1 (MCP-1) is considered a propagator of atherosclerosis and a key modulator of monocyte activity. Hormone replacement therapy (HRT) is currently being investigated as a means towards prevention of atherosclerosis. We aimed to assess (1) the range of circulating MCP-1 levels in postmenopausal women, (2) the correlation between MCP-1 and atherosclerotic burden, and (3) the effects of commencement and discontinuation of HRT on MCP-1 serum levels. METHODS: This clinical prospective trial investigated 51 postmenopausal women at increased risk for cardiovascular events who were randomized to receive either no HRT or 1 mg 17 beta-estradiol continuously plus sequential progestagen over 1 year. Intima-media thickness (IMT) of carotid and femoral arteries was measured by ultrasound. Serum levels of MCP-1 and cellular adhesion molecules were measured by ELISA. RESULTS: At baseline, MCP-1 levels and overall mean maximum IMT correlated (r=0.589; P<0.0001, Pearson's coefficient). MCP-1 levels in serum gradually decreased after 3, 6, and 12 months of HRT by 16.8 +/- 15.7% at 12 months (P<0.0001, MANOVA). Similarly, all cellular adhesion molecules decreased significantly by 6-12%. After 12 months, women decided whether to continue or discontinue treatment. At 18 months, in women discontinuing HRT (n=17), MCP-1 levels rose by 21 +/- 20% (P=0.003), but remained lowered in women continuing HRT. CONCLUSION: Our observations indicate that 17 beta-estradiol may have an antiatherosclerotic effect by reducing MCP-1 serum levels and cell adhesion molecules. PMID- 11861036 TI - Endothelium-independent effect of estrogen on Ca(2+)-activated K(+) channels in human coronary artery smooth muscle cells. AB - OBJECTIVE: Postmenopausal estrogen replacement therapy lowers the incidence of cardiovascular disease, suggesting that estrogens support cardiovascular function. Estrogens dilate coronary arteries; however, little is known about the molecular basis of how estrogen affects the human coronary circulation. The cellular/molecular effects of estrogen action on human coronary smooth muscle were investigated in the present study. METHODS: Patch-clamp and fluorescent microscopy studies were performed on human coronary myocytes in the absence of endothelium. RESULTS: Estrogen increased whole-cell currents over a range of membrane potentials, and further studies indicated that the large-conductance (186.5 +/- 3 pS), calcium- and voltage-activated potassium (BK(Ca)) channel was the target of estrogen action. Channel activity was stimulated approximately 15 fold by nanomolar concentrations of 17 beta-estradiol, and this stimulation was reversed >90% by inhibiting cGMP-dependent protein kinase activity with 300 nM KT5823. 17 beta-Estradiol increased the level of cGMP and nitric oxide in human myocytes, and the stimulatory effect of estrogen on channel activity and NO production was reversed by inhibiting NO synthase with 10 microM N(G)-monomethyl L-arginine. CONCLUSIONS: Our cellular and molecular studies identify the BK(Ca) channel as a target of estrogen action in human coronary artery smooth muscle. This response to estrogen involves cGMP-dependent phosphorylation of the BK(Ca) channel or a closely associated regulatory molecule, and further evidence suggests involvement of the NO/cGMP signaling system in coronary smooth muscle. These findings are the first to provide direct evidence for a molecular mechanism that can account for endothelium-independent effects of estrogen on human arteries, and may also help explain why estrogens reduce myocardial ischemia and stimulate coronary blood flow in patients with diseased coronary arteries. PMID- 11861037 TI - Male and female mice overexpressing the beta(2)-adrenergic receptor exhibit differences in ischemia/reperfusion injury: role of nitric oxide. AB - OBJECTIVE: Cardiac overexpression of beta(2)-adrenergic receptors (beta(2)ARs) in male mice (MTG4) results in increased contractility and increased ischemic injury. Considering recent clinical data indicating that premenopausal women are protected from cardiovascular injury, we assessed the consequences of beta(2)AR overexpression in females (FTG4). Since protection in females is mediated via estrogen, which activates endothelial and inducible nitric oxide synthases (eNOS and iNOS) we also examined the role of NOS in ischemia/reperfusion injury in male and female TG4 and wild-type (WT) mice. METHODS: Hearts from MTG4, FTG4, MWT and FWT mice were isolated and perfused in the Langendorff mode. Hearts were pretreated with either 1 micromol/l of the nonspecific NOS inhibitor, L-NAME, or 100 nmol/l of the specific iNOS inhibitor, 1400W. Control hearts received no treatment. All hearts were subjected to 20 min ischemia and 40 min reperfusion while 31P-NMR spectra were acquired. RESULTS: During ischemia, ATP and pH fell lower in MTG4 hearts than in FTG4 or WT hearts. Hearts from MTG4 mice exhibited increased ischemia/reperfusion injury as indicated by lower recoveries of postischemic contractile function, ATP and PCr than WT. Despite contractility being elevated in FTG4 hearts to the same level as MTG4 hearts, ischemia/reperfusion injury was not increased, as indicated by similar postischemic recoveries of contractile function, ATP and PCr in FTG4 hearts compared to WT. ATP and pH fell lower during ischemia in L-NAME-treated FTG4 hearts than in untreated FTG4 hearts, falling as low as untreated MTG4s. Recoveries of contractile function, ATP and PCr were as low in L-NAME-treated FTG4 hearts as in untreated MTG4 hearts and lower than untreated FTG4 hearts. In contrast, 1400W had no effect on FTG4 hearts. MTG4 hearts were unaffected by L NAME or 1400W. CONCLUSIONS: beta(2)AR overexpression increased ischemia/reperfusion injury in males but not females, thus females were protected from the detrimental effects of beta(2)AR overexpression. Protection was abolished by treatment with L-NAME, but not 1400W, implying that protection was mediated by eNOS not iNOS. PMID- 11861038 TI - Renin angiotensin system and gender differences in the cardiovascular system. AB - In the effort to explain gender-related differences of the cardiovascular system, the renin-angiotensin system experienced intensive exploration. Indeed, the development of hypertension as well as the progression of coronary artery disease and heart failure have two factors in common: (1) display distinct gender specific characteristics and (2) are enhanced by the renin-angiotensin system. It is therefore interesting to note that data from experimental animals, epidemiological surveys, and clinical investigations suggest that the components of the circulating as well as tissue-based renin-angiotensin system are markedly affected by gender. However, the issue is complicated by counter-regulatory effects of estrogen on the system with the substrate, on one hand, and the processing enzymes as well as the chief receptor, on the other hand. In fact, angiotensinogen is up-regulated particularly by oral administration of estrogen, whereas renin, angiotensin-converting enzyme (ACE), and AT-1 receptor are down regulated by the hormone. While under well-defined experimental conditions the net effect of estrogen appears to result in suppression of the renin-angiotensin system, the clinical situation may be more complex. The judgment is further complicated by the difficulty in precisely measuring the activity of the system at the tissue level. Moreover, clinically relevant read-outs for the activity of the renin-angiotensin system may be regulated multifactorially or only indirectly affected by the system. Nevertheless, the undisputable, profound biochemical changes in the renin-angiotensin system related to the estrogen status allow speculation that such interaction explains some of the differences in the cardiovascular system of men and women. PMID- 11861039 TI - Gender, sex hormones and autonomic nervous control of the cardiovascular system. PMID- 11861040 TI - Sex hormones and hypertension. AB - Gender has an important influence on blood pressure, with premenopausal women having a lower arterial blood pressure than age-matched men. Compared with premenopausal women, postmenopausal women have higher blood pressures, suggesting that ovarian hormones may modulate blood pressure. However, whether sex hormones are responsible for the observed gender-associated differences in arterial blood pressure and whether ovarian hormones account for differences in blood pressure in premenopausal versus postmenopausal women remains unclear. In this review, we provide a discussion of the potential blood pressure regulating effects of female and male sex hormones, as well as the cellular, biochemical and molecular mechanisms by which sex hormones may modify the effects of hypertension on the cardiovascular system. PMID- 11861041 TI - Estrogenic hormone action in the heart: regulatory network and function. AB - Cardiovascular diseases are the leading cause of death in the industrialised countries and display significant gender-based differences. Estrogen plays an important role in the pathogenesis of heart disease and is able to modulate the progression of cardiovascular disease. The focus on the beneficial influence of estrogen is gradually shifting from the vascular system to the myocardium. The presence of functional estrogen receptors in the myocardium has been demonstrated. Estrogen is important for cardiovascular baseline physiology and modulates the myocardial response under pathological conditions. Here we summarise the current knowledge of the regulatory network of estrogenic action in the myocardium and its effects on cardiovascular function. PMID- 11861042 TI - Gender-specific differences of cardiac remodeling in subjects with left ventricular dysfunction: a population-based study. AB - BACKGROUND: Recent studies suggest that female gender is associated with a lower prevalence and a more benign prognosis of heart failure. In the current population-based study, it was our objective to evaluate the implications of gender on the association between impaired left ventricular (LV) function and mass as well as neurohumoral activation. METHODS AND RESULTS: A total of 1883 subjects (992 female, 891 male) of two MONICA surveys in Augsburg, Germany, were analyzed. Participants of one of these surveys were additionally characterized with respect to neurohormonal activation. As compared to men, women were characterized by a slightly higher LV ejection fraction (EF, Teichholz-Method, 65.4 +/- 0.3% vs. 63.4 +/- 0.3, P<0.01) and a markedly lower LV mass index (LVMI 81 +/- 1 g/m(2) vs. 96 +/- 1, P<0.01). As compared to men with normal LV function, those with LV dysfunction (EF below mean minus two standard deviations, S.D.) were characterized by significantly increased LV mass (LVMI +48%, P<0.01), plasma BNP (+373%, P<0.01) and ANP (+57%, P<0.01), while no significant changes were observed in women (LVMI +3%, BNP +48%, ANP +27%, all P=n.s). Only a small subgroup of women with severe LVD (EF below mean - 3 S.D.) was characterized by significantly increased LV mass (LVMI +23%, P<0.05 vs. control and LVD), however, this increase was less pronounced as compared to men with severe LVD (LVMI +46%, P<0.01 vs. control). Gender-specific differences between LV function and structure were also confirmed by multivariate analysis. While LVMI was independently and significantly correlated with EF in male subjects in addition to systolic blood pressure, age, and body mass index (all P<0.01), these parameters displaced EF as a predictor of LVMI in female subjects. CONCLUSIONS: Men with moderate or severe LV dysfunction are characterized by an increase in both LV mass and cardiac natriuretic peptide plasma concentrations. In contrast, LV mass and natriuretic peptide concentrations increase to a lesser extent and only with severe LV dysfunction in women. These observational data suggest gender specific control of myocardial adaptations to hemodynamic overload and a more rapid induction of LV hypertrophy during myocardial dysfunction in male subjects. PMID- 11861043 TI - Ovarian hormones induce TGF-beta(3) and fibronectin mRNAs but exhibit a disparate action on cardiac fibroblast proliferation. AB - Prior to menopause, women have a lower risk of cardiovascular disease compared to age-matched men. Despite the well-documented beneficial physiological effects of ovarian hormones on vascular reactivity and growth, very little is known with regard to the direct action on cardiac cells. OBJECTIVE: The following study examined the pattern of ovarian hormone receptor subtype expression in cardiac fibroblasts, the modulator role of 17 beta-estradiol and progesterone on growth and their respective influence on putative molecular events of extracellular matrix remodeling. METHODS AND RESULTS: Neonatal rat cardiac fibroblasts were isolated from 1- to 3-day-old Sprague--Dawley rats. Immunofluorescence and Western blot analysis revealed the presence of estrogen receptor-alpha (ER alpha), and -beta (ER-beta) subtypes, with the ER-alpha subtype localized on the plasma membrane. Likewise, both progesterone receptor-A (PR-A), and -B (PR-B) subtypes were expressed in cardiac fibroblasts, and the PR-B appeared to be the predominant subtype associated with the plasma membrane. Despite the presence of both ER subtypes, the treatment of cardiac fibroblasts with 1 microM 17 beta estradiol exerted a modest decrease in DNA synthesis. By contrast, progesterone treatment caused a dose-dependent decrease in [3H]thymidine uptake, without a concomitant induction of apoptosis. The progesterone-mediated decrease in DNA synthesis was associated with the upregulation of the cyclin-dependent kinase inhibitor p27(Kip1), whereas p21(cip) and proliferating cell nuclear antigen protein levels were unchanged. Lastly, despite the modest effect on DNA synthesis, 17 beta-estradiol increased the steady-state mRNA levels of transforming growth factor-beta(3) and fibronectin. Likewise, progesterone increased the expression of both transforming growth factor-beta(3), and fibronectin mRNA. CONCLUSION: Collectively, these data are the first to highlight the presence of estrogen and progesterone receptor subtypes on the plasma membrane of neonatal rat cardiac fibroblasts, and further underscore the ability of ovarian hormones to directly suppress DNA synthesis, and influence putative molecular events associated with extracellular matrix remodeling. PMID- 11861044 TI - Sex, hormones, and repolarization. AB - There is increased awareness of the impact of gender and gonadal steroids on human cardiac rhythm and arrhythmias; e.g., drugs that prolong repolarization induce torsades de pointes (TdP) more frequently in women than men; female gender is an independent risk factor for syncope and sudden death in the congenital long QT syndrome; and the higher propensity toward arrhythmia in normal females is associated with fundamental differences in repolarization such that rate corrected QT intervals are longer in females than males. Mechanisms underlying these differences are incompletely defined but are believed to involve gonadal steroids. This review discusses recent advances and prospects for further elucidation of the influence of gender and gonadal steroids on ventricular repolarization and arrhythmias. PMID- 11861046 TI - Reduced repolarization reserve in ventricular myocytes from female mice. AB - OBJECTIVE: Cardiac repolarization is prolonged and repolarization reserve (RR) is diminished in female rabbits and humans, compared to males. Reduced RR is evidenced by the relatively greater increase in ventricular action potential duration (APD) in myocytes from females in response to drugs that block repolarizing K(+) currents. Mice are an increasingly important experimental model animal for cardiovascular research, but gender-dependent differences have not been reported for repolarization in murine ventricular myocytes. METHODS: APD and repolarizing K(+) currents were measured in isolated ventricular myocytes from adult littermate male and female mice. Repolarizing K(+) currents were dissected into transient (I(to)) and sustained (I(sus)) components and the selective I(sus) antagonist FK506 was used to probe for differences in RR. RESULTS: Under control conditions APD at 50% (APD(50)) and at 90% (APD(90)) repolarization was significantly longer in females (APD(50)=15 +/- 3 ms, n=6 and APD(90)=63 +/- 6 ms, n=6) compared to males (APD(50)=8 +/- 2 ms, n=7 and APD(90)=42 +/- 9 ms, n=7) at 1.0 Hz. At 0.3 Hz stimulation frequency APD(90), but not APD(50), was significantly longer in females (APD(50)=12 +/- 2 ms and APD(90)=54 +/- 5 ms, n=10) compared to males (APD(50)=11 =/- 2 ms and APD(90)=47 +/- 7 ms, n=10). FK506 treatment (25 microM) selectively and equally inhibited I(sus) in all cells, and significantly increased APD(50) and APD(90) in males and females at 0.3 and 1.0 Hz. However, increases in APD(50) and APD(90) (0.3 and 1.0 Hz) in response to FK506 were significantly greater in myocytes from females compared to males. Voltage clamp measurement of I(to) and I(sus) revealed that males had a relatively more prominent I(to) while females exhibited a more prominent I(sus). CONCLUSIONS: Ventricular action potential repolarization is prolonged in myocytes from female compared to male mice. Female mice have reduced RR that is unmasked by FK506. These findings suggest that gender is an important variable for cardiovascular studies using mice. PMID- 11861045 TI - Effects of gonadal steroids on gender-related differences in transmural dispersion of L-type calcium current. AB - OBJECTIVES: Repolarization-prolonging drugs induce torsades de pointes (TdP) in females more than males. The action potential plateau and the early afterdepolarizations that induce TdP are determined, in part, by L-type calcium current (I(Ca,L)). Therefore, we studied gender- and hormone-related differences in I(Ca,L) in age-, and weight-matched normal male, female and hormonally treated, castrated rabbits. METHODS: Oophorectomized (OVX) or orchiectomized (ORCH) 50- to 60-day-old rabbits were subcutaneously implanted with pellets impregnated with placebo (PLA), 5 alpha-dihydroxytestosterone (DHT), or 17 beta estradiol (EST). Four to five weeks later, epicardial and endocardial myocytes were isolated from the left ventricle. Patch clamp technique was performed to assess I(Ca,L). RESULTS: I(Ca,L) density (measured as peak current density [pA/pF] at +15 mV, V(h)= -40 mV), was greater in female epicardium (-7.4 +/- 0.9) than endocardium (-5.6 +/- 0.7, P<0.05), while male epicardial I(Ca,L) density ( 6.5 +/- 0.7) did not differ from endocardial (-5.9 +/- 1.0, P>0.05). OVX-female, DHT and EST-treated groups had epicardial I(Ca,L) density (-5.6 +/- 0.6, and -5.9 +/- 0.7, respectively) greater than endocardial (-4.3 +/- 0.3, and -3.6 +/- 0.4, P<0.05). However, OVX-females had hormone levels not significantly different from female controls and EST-treated females had non-physiological levels of estradiol. There were no differences between endocardial and epicardial I(Ca,L) activation and inactivation. In contrast, epicardial-endocardial differences in I(Ca,L) density in EST-treated OVX-females were associated with epicardial endocardial differences in I(Ca,L) activation and conductance; in DHT-treated OVX females only epicardial-endocardial activation differed. The other groups, showed no I(Ca,L) transmural gradient, or differences in activation, inactivation or conductance. CONCLUSIONS: The greater dispersion in I(Ca,L) density of OVX-DHT and OVX-EST than OVX-PLA suggests both hormones can modulate I(Ca,L) density in females. That gonadal steroids had no effect on I(Ca,L) dispersion in males suggests gender differences in mechanism of action of both hormones. The greater I(Ca,L) dispersion in females may contribute to gender differences in repolarization. PMID- 11861047 TI - Gender differences in the long QT syndrome: effects of beta-adrenoceptor blockade. AB - BACKGROUND: Gender differences have been reported in patients with the congenital long QT syndrome (LQTS). We analyzed whether electrocardiographic differences existed in females, males, girls and boys in response to beta-adrenoceptor blockade. METHODS: 12-lead ECGs before and during beta-adrenoceptor blockade were collected in 87 genotyped LQTS patients (48 women, 14 men, 12 girls and 13 boys). Up to three QTc intervals were determined in each lead of the ECG. V4 was used for QT/QTc analysis. Difference between longest and shortest QT interval was taken as a measure for dispersion of QT intervals. RESULTS: (1) Adult males had the greatest shortening of the QTc interval upon treatment with beta-adrenoceptor blockade. During treatment, adult males with LQTS(1) (mutation in the KCNQ1 gene, affecting I(Ks) current) were found to have shorter QTc intervals than adult females; this difference did not exist in LQTS(2) patients (mutation in the HERG gene, affecting I(Kr) current). (2) Female LQTS(2) patients had a 50% larger dispersion than female LQTS(1) patients both before and during treatment. (3) Adult male LQTS(1) patients constitute the only patient group with a marked decrease in QTc intervals and dispersion associated with a 100% efficacy of treatment in response to beta-adrenoceptor blockade. CONCLUSIONS: These findings indicate that, in addition to underlying differences in repolarization between men and women, cardiac electrophysiological responses to beta-adrenoceptor blockade can be modulated by gender-related factors. PMID- 11861048 TI - Implantable contraceptives for women. PMID- 11861049 TI - Toxicology of progestogens of implantable contraceptives for women. AB - There are currently four progestogens used in implantable contraceptives marketed or tested in clinical trials: levonorgestrel in Norplant and Jadelle, etonogestrel (3-keto-desogestrel) in Implanon, nestorone in Elcometrine, and nomegestrol acetate in Uniplant and Surplant. Each progestogen was evaluated for hormonal activity and for safety in a wide variety of tests in vitro and in animals prior to their use in women. All four progestogens underwent pre-clinical testing that generally followed the format for animal testing of steroidal contraceptives published by the World Health Organization and the US Food and Drug Administration (FDA). Most of the progestogens have been tested for genotoxicity in bacterial and mammalian cultured cells and in rodents. All were tested for toxicity in short- and long-term toxicology studies in rodents and dogs or monkeys, and all were tested for their effects on reproduction and fetal development. In most cases, the progestogens were tested for carcinogenicity in two rodent species, rats and mice. Early clinical trials in small numbers of women provided additional safety data prior to the exposure of large numbers of women in Phase 3 clinical trials. The published data and data submitted to the FDA demonstrate that the implantable progestogens have no significant or unusual toxicities and have a similar safety profile to the progestogens found in the approved oral contraceptives. PMID- 11861050 TI - Toxicology of polymers for implant contraceptives for women. AB - This article reviews the toxicology of polymers that are used in contraceptive implants. The two main classes of synthetic, nondegradable polymers used in the delivery of female contraceptives are silicone elastomers (e.g., Silastic) and ethylene co-vinyl acetate (EVA; ELVAX). The controversies surrounding the silicone breast implants have prompted several studies to evaluate the toxicity of silicones. The epidemiologic data obtained thus far have overwhelmingly concluded that no correlation exists between certain chronic symptoms, such as arthritis, in patients and silicone prosthesis. This conclusion has been echoed by the expert panel report by the Institutes of Medicine. Although the IOM report focused on the safety of silicone breast prosthesis, data emerging from the joint reconstruction area also suggests that Silastic is safe for in vivo use. The toxicological studies on EVA are few, and the conclusion thus far is that they elicit no adverse local or systemic response over extended periods in vivo. In conclusion, the prognosis for Silastic and ELVAX as of now is excellent. However, any future implant development using these polymers should place an emphasis on processing parameters to minimize potential small molecule leachants and establish safety as a function of both site and duration of implantation. PMID- 11861051 TI - Progestin implants for female contraception. AB - Four different implants, in the form of capsules or covered rods, that release one of the synthetic progestins levonorgestrel, etonogestrel, Nestorone, or Elcometrine and nomegestrol acetate were reviewed. Biocompatible polymers or copolymers of polydimethyl/polymethylvinyl-siloxanes or ethylvinylacetate are used to hold the steroid crystals and to control the rate of release. Once inserted under the skin, these implants release the corresponding steroid continuously over prolonged periods, a process that can be readily interrupted by implant removal. During long-term use of the implant, the released steroid circulates in blood at a fairly stable level. The physical characteristics of the implants, including drug contents and rate of release, serum levels of the progestin during use, and the duration of their effective life are described. Total steroid loads vary in the range of 50 mg to 216 mg; average release rates are in the range of 30-100 ug/day, and effective lives from 6 months to 7 years. PMID- 11861052 TI - Mechanisms that explain the contraceptive action of progestin implants for women. AB - Four different contraceptive implants for women, in the form of capsules or covered rods, that release either one of the synthetic progestins levonorgestrel, etonogestrel, Nestorone, or Elcometrine and nomegestrol acetate were considered. These progestins act by binding to their receptors located in diverse target cells, which are distributed along the hypothalamic-pituitary-gonadal-genital tract axis. These implants differ in the extent to which each one interferes with various steps of the reproductive process and in the intensity with which each one affects the same process along its effective life, but they have in common the ability to interfere with several key processes required for gamete encounter and fertilization. The steps they interfere with most are the ovulatory process, through partial or complete inhibition of the gonadotropin surge, and by changing the quality of cervical mucus; they restrict or suppress the access of fertile spermatozoa to the site of fertilization. Changes in endometrial development also occur, but this contribution to the contraceptive action is difficult to determine at the present time. PMID- 11861053 TI - Implantable contraceptives for women: effectiveness, discontinuation rates, return of fertility, and outcome of pregnancies. AB - Progestogen-only contraceptive implants are highly effective. In most studies, 5 year cumulative pregnancy rates are less than 1.5/100 women for Norplant and Norplant II. No study has yet reported any failures with Implanon. Weight over 70 kg and age at insertion under 25 years both increase the failure rate of Norplant and Norplant II; however, data are as yet lacking for Implanon. The effectiveness of other progestogen-only implants for which there are as yet few data are unlikely to be any different. Continuation rates are high compared with other hormonal methods and with the intrauterine device. In most cohorts at least 35% of women, and often many more, are still using Norplant by the end of 5 years. Rates vary according to a number of factors, including population studied, age, and parity. Menstrual disturbance is by far the most common reason for discontinuation, with headache, acne, weight gain, and desire for pregnancy accounting for other common reasons for implant removal. Fertility returns rapidly following implant removal, and pregnancy rates (76-100% 1 year after removal) are usually no different from those following discontinuation of any other contraceptive method. There is no increase in the risk of ectopic pregnancy, fetal malformation, or impaired infant health in pregnancies conceived either during implant use or after removal. PMID- 11861054 TI - Contraceptive implants and lactation. AB - The safety and efficacy of four contraceptive implants, plant, Implanon, Nestorone and Elcometrine, have been evaluated during use in the postpartum period by lactating women. These implants provide highly effective contraceptive protection with no negative effect on breastfeeding or infant growth and development. Breastfeeding women initiating Norplant use in the second postpartum month experience significantly longer periods of amenorrhea than do untreated women or intrauterine device users. After weaning, the bleeding pattern is similar to that observed in non-nursing women. Norplant use does not affect bone turnover and density during lactation. Norplant and Implanon release orally active progestins while Nestorone and Elcometrine implants release an orally inactive progestin, which represents an advantage since the infant should be free of steroidal effects. The infant's daily intake of steroids (estimated from concentrations in maternal milk during the first month of use) range from 90 to 100 ng of levonorgestrel (Norplant), 75-120 ng of etonogestrel (Implanon), and 50 ng and 110 ng of Nestorone (Nestorone and Elcometrine implants, respectively). Nursing women needing contraception may use progestin-only implants when nonhormonal methods are not available or acceptable. Implants that deliver orally active steroids should only be used after 6 weeks postpartum to avoid transferring of steroids to the newborn. PMID- 11861055 TI - Metabolic effects of implantable steroid contraceptives for women. AB - The metabolic impact of progestin-only contraceptives is less than that of combined oral contraceptives. Subdermal contraceptive implant systems that provide a sustained release of low levels of progestins are now becoming widely available. This review evaluates the metabolic effects of currently available products that release the progestins levonorgestrel (Norplant, Jadelle, and their Chinese equivalents); etonogestrel (Implanon); nomegestrol acetate (Uniplant); and Nestorone, formally called ST-1435 (Nestorone implant/Elcometrine). Data on liver, kidney, and renal function; carbohydrates and insulin release; hemostasis; blood pressure; and lipids are considered. The metabolic effects reported for these methods as a whole were minimal. Any changes were generally within the normal range for the populations studied and, therefore, are unlikely to be of clinical significance. However, all published studies have been conducted in healthy populations of women. To inform clinical practice, the field would be well served to have additional empiric data from well-designed, well-implemented, and well-reported trials in women who are deemed to be at elevated risk for certain diseases including cardiovascular disease and diabetes. PMID- 11861056 TI - Nonmenstrual adverse events during use of implantable contraceptives for women: data from clinical trials. AB - Contraceptive methods, including implants, do not prevent common symptoms and adverse health events that most people experience. It is difficult, therefore, to decide whether or not the occurrence of symptoms or adverse events that are common can be attributed to use of a contraceptive method or to determine if a given method changes the likelihood of their occurrence. Based on the review of the literature, no apparent differences in the frequency of adverse events are evident between the six-implant or two-rod levonorgestrel systems and the single implant etonogestrel and nomegestrol acetate systems. The most frequent adverse events reported in clinical trials that are probably related to implant use are headaches and acne. Weight gain, dizziness, and mood changes are also frequently mentioned adverse events and are possibly steroid-related. Other possibly related adverse events, although much less frequently reported, are loss of libido, fatigue, hair loss, and other skin conditions. Persistent ovarian follicles that spontaneously disappear are a common event during use of progestin-only contraceptives, and providers should be aware of this condition to avoid unnecessary interventions. Overall, the vast experience reported in the clinical studies reviewed here show that all existing implantable contraceptives are equally safe. This can probably be attributed to the low-hormonal dose delivered by progestin-implant systems. PMID- 11861058 TI - Safety of implantable contraceptives for women: data from observational studies. AB - Contraceptive implants are registered in over 60 countries and have been used by millions of women for three decades. This article reviews findings from observational studies on the safety of contraceptive implants and examines the risk of specific health outcomes. Fifty-five articles were reviewed, and the body of evidence for each health outcome was summarized. Available evidence suggests that contraceptive implants are safe and, overall, implant users do not experience adverse events at rates higher than women not using implants. With respect to specific outcomes, the evidence suggests no increased risks of pelvic inflammatory disease, decreased bone mineral density, anemia, thrombocytopenia, or death with implant use. The evidence was too limited to draw meaningful conclusions for neoplastic disease, cardiovascular events, and HIV/AIDS. Nonsignificantly elevated associations were reported for diabetes, serious mental disorders, and rheumatoid arthritis. Conditions for which risks were marginally, yet significantly, elevated were hypertension and gall bladder disease. PMID- 11861057 TI - Vaginal bleeding disturbances and implantable contraceptives. AB - Implantable contraceptives allow safe and effective fertility regulation for up to 5 years. Currently available devices release low doses of progestogens. Disruption of vaginal bleeding patterns is almost inevitable, particularly during the initial months of use. Irregular and prolonged bleeding as well as amenorrhea are common. Irregular bleeding is unpopular with most women and unacceptable to some. This review describes the vaginal bleeding disturbances induced by modern implantable contraceptives and discusses the implications of these in terms of method use and discontinuations. The cause of the irregular bleeding is not fully understood, but recent evidence suggests that an increase in endometrial vascular fragility might precipitate vessel breakdown and, hence, breakthrough bleeding. This review discusses this evidence and outline the possible mechanisms underlying breakthrough bleeding associated with implantable contraceptives. In addition, therapies for bleeding disturbances are described and their efficacy reviewed. PMID- 11861059 TI - Health services at the clinic level and implantable contraceptives for women. AB - The quality of implant service provision, particularly counseling, has been associated with successful use and with fewer discontinuations for side-effects. Requirements necessary for quality service provision include cadres of health care workers who can provide implants, training curriculum, duration of training, and training techniques; knowledge of the facilities, surgical equipment, and other supplies necessary; infection prevention steps to safely provide implants; techniques for managing side-effects; methods for managing difficult implant removals, the importance of maintaining close relationships with implant clients, and establishing communication and notification systems for removal (and sometimes replacement) when the effective life-span of the implants has been reached. In this article we review the components and training necessary for the establishment and maintenance of quality implant service delivery systems, discuss the implications of providing more than one type of implant, and describe trends in use. PMID- 11861060 TI - Users' perspectives on implantable contraceptives for women. AB - Users of subdermal contraceptive implants report long duration of use, convenience, and high efficacy as liked features and bleeding irregularities, weight loss/gain, and headaches as disliked effects of these methods. Satisfied users tend to be women who do not experience side effects or women who value positive features and tolerate the side effects. Satisfaction of the users is also closely related to quality of services. Service delivery problems, such as lack of proper pre-insertion counseling, inadequacy in number and imbalance in the geographical distribution of providers who can insert and remove implants, and appropriately manage side effects have been observed in new programs. Insensitivity of some providers to side-effects, such as bleeding irregularities, and to requests for removal have caused user dissatisfaction and serious public concern. Care must be taken to provide implants in a context of high quality of care with adequate numbers and distribution of trained providers and a basic record-keeping and tracking system. PMID- 11861061 TI - Recent developments in contraceptive implants at the Population Council. AB - Development of contraceptive implant methods, when based on established or on new synthetic chemical entities, is a decadal or multi-decadal process. The process often requires the cooperation of numerous investigators for laboratory work, for early Phase II trials, for dose-finding trials, and for Phase III clinical trials. The Phase III work also requires cooperation with a commercial manufacturer and potential distributor of the product. The Population Council has recently completed developmental work on two levonorgestrel-releasing implants, with filings to regulatory agencies that support extended use of Jadelle implants for 5 years and Norplant implants for 7 years. When the developmental process includes establishing the clinical properties of a molecule not yet approved by regulatory agencies, the minimum development time appears to be two decades. The status and rationale of studies of a new Nestorone-releasing, single implant developed by the Population Council for a period of use of 2 years are presented. PMID- 11861062 TI - Myeloma expression systems. AB - Myeloma expression systems have been utilized successfully for the production of various recombinant proteins. In particular, myeloma cell lines have been exploited to express a variety of different antibodies for diagnostic applications as well as in the treatment of various human diseases. The use of myeloma cells for antibody production is advantageous because they are professional immunoglobulin-secreting cells and are able to make proper post translational modifications. Proper glycosylation has been shown to be important for antibody function. Advances in genetic engineering and molecular biology techniques have made it possible to isolate murine and human variable regions of almost any desired specificity. Antibodies and antibody variants produced in myeloma cells have been extremely helpful in elucidating the amino acid residues and structural motifs that contribute to antibody function. Because of their domain nature, immunoglobulin genes can be easily manipulated to produce chimeric or humanized antibodies. These antibodies are less immunogenic in humans and also retain their specificity for antigen and biologic properties. In addition, novel proteins in which antibodies are fused to non-immunoglobulin sequences as well as secretory IgA have been produced in myeloma cells. PMID- 11861063 TI - Biological assays for interferons. AB - Interferons (IFN) are potent biologically active proteins synthesised and secreted by somatic cells of all mammalian species. They have been well characterised, especially those of human origin, with respect to structure, biological activities, and clinical therapeutic effects. While structural differences are known to exist among the IFN species that constitute the "IFN family" and despite the existence of different receptors for type I and type II IFN, all species have been shown to exert a similar spectrum of in vitro biological activities in responsive cells. Principal among the biological activities induced by IFN is antiviral activity, the activity used to originally define IFN. Antiviral activity of IFN is mediated via cell receptors and is dependent on the activation of signalling pathways, the expression of specific gene products, and the development of antiviral mechanisms. Sensitivity of cells to IFN-mediated antiviral activity is variable, and depends on a number of factors including cell type, expression of IFN receptors and downstream effector response elements, effectiveness of antiviral mechanisms, and the type of virus used to infect cells. Nevertheless, by the judicious use of sensitive cell lines in combination with appropriate cytopathic viruses, effective assays to measure the antiviral activity have been developed. Historically, "antiviral assays" (AVA) were the first type of biological assays that were developed to measure the relative activity or potency of IFN preparations. However, the subsequent discoveries of several other biological activities of IFN has opened the way to the development of assays based on one or other of these activities. The latter include inhibition of cell proliferation, regulation of functional cellular activities, regulation of cellular differentiation and immunomodulation. More recently, the cloning of IFN responsive genes has led to the development of "reporter gene assays". In this case, the promoter region of IFN responsive genes is linked with a heterologous reporter gene, for example, firefly luciferase or alkaline phosphatase, and transfected into an IFN-sensitive cell line. Stably transfected cell lines exposed to IFN increase expression of the reporter gene product in direct relation to the dose of IFN, the readout being a measure of this product's enzymic action. The current review aims to give a critical overview of the development, specificity, standardisation and present use of the various biological assay methods now available for the quantification of IFN activity. PMID- 11861064 TI - Novel strategy for the selection of human recombinant Fab fragments to membrane proteins from a phage-display library. AB - Traditionally, the selection of phage-display libraries is performed on purified antigens (Ags), immobilized to a solid substrate. However, this approach may not be applicable for some Ags, such as membrane proteins, which for structural integrity strongly rely on their native environment. Here we describe an approach for the selection of phage-libraries against membrane proteins. The envelope glycoproteins (Env) of the Human Immunodeficiency Virus type-1 (HIV-1) were used as a model for a type-1 integral membrane protein. HIV-1IHI Env, expressed on the surface of Rabbit Kidney cells (RK13) with a recombinant vaccinia virus (rVV), was solubilized using the non-ionic detergent n-Octyl beta-D-glucopyranoside (OG). Membrane associated Env was reconstituted into vesicles by the simultaneous removal of detergent and free monomeric Env subunits by gel-filtration. The resulting antigen preparation, termed OG-P1IHI, was captured on microtiter plates coated with Galanthus nivalis agglutinin (GNA) and used for rounds of selection (panning) of a well-characterized phage-display library derived from an HIV-1 seropositive donor. Simultaneously, an identical experiment was performed with OG P1IHI vesicles disrupted by Nonidet P-40 (NP-P1IHI). Both membrane-associated and soluble Ags were selected for vaccinia-specific clones (OG-P1IHI: 59/75 and NP P1IHI: 1/75) and HIV-1-specific clones (OG-P1IHI: 11/75 and NP-P1IHI: 65/75) using our approach. Hence, the novel panning strategy described here may be applicable for selection of phage-libraries against membrane proteins. PMID- 11861065 TI - Generation of functional and mature dendritic cells from cord blood and bone marrow CD34+ cells by two-step culture combined with calcium ionophore treatment. AB - The object of this study is to explore a culture method to generate a large number of functional and mature dendritic cells (DC) from human CD34+ hematopoietic progenitor cells. In the present study, we used a two-step method combined with calcium ionophore to induce DC from cord blood (CB) or normal human bone marrow (BM) CD34+ progenitor cells. The two-step method consists of 10 days of first step culture for the expansion and proliferation of CD34+ hematopoietic progenitor cells in the presence of SCF, IL-3, IL-6, G-CSF, and 7--11 days of second step culture for the induction of DC in the presence of GM-CSF, IL-4 and TNF-alpha. By the two-step culture, total nucleated cells were increased 208+/-66 (+/-SD, n=13), or 94+/-29 (n=5)-fold in the culture of CB or BM cells, respectively, compared with the number of CD34+ cells at the time of starting culture. Out of the total nucleated cells, 23 +/-10.4% of cells in CB cell culture and 25 +/-5% of cells in the BM cell culture acquired DC characteristic phenotypes, which were marked expressions of CD1a, HLA-DR, co-stimulatory molecules such as CD80, CD40, and adhesion molecule such as CD58. In allogeneic mixed leukocyte reaction (MLR), two-step cultured cells showed potent allo stimulatory capacity. With this two-step culture, the absolute number of CD1a+ cells that co-expressed HLA-DR, CD80, CD40 and CD58 was enhanced approximately 3 times in CB cell culture and 1.9 times in BM cell culture, compared with the commonly used one-step culture method for the generation of DC from CD34+ cells using SCF, GM-CSF and TNF-alpha. However, on these DC generated in the two-step culture, the expressions of co-stimulatory molecule CD86 and mature DC marker CD83 were not sufficient. By the treatment of two-step cultured cells with calcium ionophore agent (A23187), the expression of co-stimulatory molecules such as CD86 and CD80 (especially CD86) was up-regulated. Besides, the expression of mature DC marker CD83 was remarkably induced by treatment with A23187 for a short duration (24 h). Consistent with the up-regulation of surface molecules CD86, CD80 and CD83, the two-step cultured cells treated with A23187 also showed a stronger allo-stimulatory capacity compared with the cells without A23187 treatment. In conclusion, the present study demonstrated that the two-step culture method effectively improved the yield of CD1a+ DC generated from CD34+ cells, and the phenotypes and functions of these CD1a+ DC could be enhanced efficiently by treatment with a calcium ionophore agent. PMID- 11861066 TI - Phage library panning against cytosolic fraction of cells using quantitative dot blotting assay: application of selected VH to histochemistry. AB - Comprehensive preparations of antibodies against various kinds of proteins in cells would be useful in proteome research and antibody-based research. Here we report the panning of a human antibody heavy chain variable domain (VH) phage library against a cytosolic fraction of rat liver to obtain antibodies specific for certain cytoplasmic proteins. Rat liver specimens were homogenized and subjected to differential centrifugation. A 125000 x g supernatant (rat liver cytosol, RLC) was immobilized onto a nitrocellulose membrane and subjected to phage VH library panning. For efficient assessment of binding phages, we established a system that was a combination of monoclonal phage ELISA and quantitative dot blotting of phages. The VH genes of the binding phages were selected and expressed as VH--bacterial alkaline phosphatase (PhoA) conjugates (VH/RLC--PhoAs) in Escherichia coli. One of the VH/RLC--PhoAs stained one major band on Western blotting of RLC and also stained the cytoplasm of hepatocytes histochemically. This is the first report of phage library panning against the cytosolic fraction of cells to obtain human VH fragments, and the application of those human VH fragments to histochemical study. PMID- 11861067 TI - Characterization of a unique human single-chain antibody isolated by phage display selection on membrane-bound mosquito midgut antigens. AB - The insect midgut is the primary site for food digestion, as well as for vector borne pathogen infection into the invertebrate host. Accordingly, antigens of this critical insect organ are targets for anti-vector vaccines, insecticidal toxins, and transmission-blocking vaccines. We used midgut proteins of the African malaria vector mosquito Anopheles gambiae to select single-chain human antibody fragments (scFv) from a high-diversity, phage-displayed library. Using a phage-display selection method on western-blotted antigens, we selected an unusual truncated scFv clone, consisting of a heavy-chain only, which binds to An. gambiae midgut tissue. This clone binds a spectrum of mosquito antigens from the midgut and other mosquito tissues, as well as various mammalian glycoproteins, but binding was reduced when these glycoproteins were enzymatically deglycosylated. We also observed that this clone preferentially binds the lumenal midgut surface. Furthermore, antigen binding by our selected scFv was limited by competition with increasing concentrations of certain soluble carbohydrates, most dramatically by galactose and N-acetyl glucosamine. Our results show that the cognate epitope of this scFv is a carbohydrate moiety. This paper describes a phage-display selection of antibody fragments on mosquito midgut tissue and it also describes a method for phage-display selection on membrane-immobilized heterogeneous antigens. These selection methods resulted in the isolation of a novel, truncated, carbohydrate-binding human antibody fragment from a naive phage-display library. PMID- 11861068 TI - Preservation of the pattern of tyrosine phosphorylation in human neutrophil lysates. II. A sequential lysis protocol for the analysis of tyrosine phosphorylation-dependent signalling. AB - In stimulated neutrophils, the majority of tyrosine phosphorylated proteins are concentrated in Triton X-100 or NP-40 insoluble fractions. Most immunobiochemical studies, whose objective is to study the functional relevance of tyrosine phosphorylation are, however, performed using the supernatants of cells lysed in non-ionic detergent-containing buffers (RIPA lysis buffers). This observation prompted us to develop an alternative lysis protocol. We established a procedure involving the sequential lysis of neutrophils in buffers of increasing tonicities that not only preserved and solubilized tyrosine phosphorylated proteins but also retained their enzymatic activities. The sequential lysis of neutrophils in hypotonic, isotonic and hypertonic buffers containing non-ionic detergents resulted in the solubilisation of a significant fraction of tyrosine phosphorylated proteins. Furthermore, we observed that in monosodium urate crystals-stimulated neutrophils, Lyn activity was enhanced in the soluble fraction recovered from the hypertonic fraction, but not from that of the first hypotonic lysis. The distribution of tyrosine phosphorylated proteins between the NP-40 soluble and insoluble fractions was both substrate- and agonist-dependent. In neutrophils stimulated with fMet-Leu-Phe, MSU crystals or by CD32 ligation, the tyrosine phosphorylated proteins were mostly insoluble. On the other hand, in GM-CSF-treated cells, the phosphoproteins were more equally distributed between the two fractions. The results of this study provide a new experimental procedure for the investigation of tyrosine phosphorylation pathways in activated human neutrophils which may also be applicable to other cell types. PMID- 11861069 TI - Analysis of the binding of bispecific monoclonal antibodies with immobilized antigens (human IgG and horseradish peroxidase) using a resonant mirror biosensor. AB - The interaction between two monoclonal antibodies (mAbs) and their corresponding bispecific antibody (bAb) with immobilized antigens has been examined using a resonant mirror biosensor (IAsys). BAbs were produced by cell fusion. The analysed panel of affinity-purified antibodies included two parental mAbs, one specific to human IgG (hIgG), and another specific to horseradish peroxidase (HRP), and a bAb derived thereof (anti-hIgG/HRP). The real-time analysis showed the drastic differences in the avidity of bivalent anti-HRP mAbs and anti-HRP shoulder of bAbs. Thus, the observed equilibrium association constant (K(ass)) of anti-HRP mAbs was about 50 times higher that of anti-HRP shoulder of bAbs. The ratio of association rate constants (k(ass)) of mAbs and bAbs was about two, due to the statistical factor of two binding sites per bivalent antibody molecule. However, the dissociation rate constant (k(diss)) of anti-HRP shoulder of bAbs was 21 times higher k(diss) of anti-HRP mAbs. The comparison with the theoretical model shows that these observations are consistent only with a situation in which bivalent binding of mAbs with immobilized HRP predominates over monovalent binding. On the contrary, the second parental mAb (anti-hIgG) did not show the increase in avidity due to bivalent binding, compared to the anti-hIgG shoulder of bAbs, suggesting that this mAb was bound monovalently to immobilized hIgG. The K(ass) values determined by solid-phase radioimmunoassay (RIA) yielded figures almost overlapping with those obtained by IAsys. The results of the comparison of bAbs and mAbs are discussed from the viewpoint of the use of bAbs in heterogeneous systems. On the other hand, these data demonstrate that real-time analysis of antibody binding parameters in IAsys biosensor is valuable for the selection of mAbs and bAbs with desired features, for different fields of application. PMID- 11861070 TI - A mixed hemadsorption assay for detection of cell surface binding anti-tumor antibodies in human sera. AB - A practical mixed hemadsorption assay (MHA) was developed for the detection of cell surface binding anti-tumor antigen antibodies (anti-TA Ab) in the human sera. The assay was compared to enzyme-linked immunosorbent assay (ELISA) and dot ELISA in the detection of the antibodies developed against GA733-2 antigen. SW1116 cell line and recombinant GA733-2E protein were used as targets in the MHA and solid phase immunoassays (SPIA), respectively. Sera were obtained from healthy donors and vaccinated-colon carcinoma patients. The sensitivity level of the MHA was very high for detection of anti-TA Ab in sera. PMID- 11861071 TI - Versatility of merocyanine 540 for the flow cytometric detection of apoptosis in human and murine cells. AB - Since apoptosis plays many roles in development, immune function, and disease, there is an ongoing need to identify inexpensive and reliable fluorochromes for the quantitation of apoptosis. Merocyanine 540 (MC540) binds to the outer membrane of cells and readily fluoresces in the highly disordered membranes of apoptotic cells making them readily detectable by flow cytometry. Protocols for the effective labeling and gating of MC540br apoptotic cells are provided. For example, MC540br cells from dexamethasone (Dex) treated thymocytes were found to be equivalent in proportion to apoptotic cells noted in the propidium iodide (PI) stained and annexin-V stained populations. Sorting of the MC540br cells followed by counterstaining with PI demonstrated that these cells resided in the low DNA fluorescent or sub-G1 region and were small in size based on light scatter. Dexamethasone, etoposide, irradiation, and a calcium ionophore were used to induce cell death with equivalent numbers of apoptotic cells obtained with MC540 and PI. Moreover, apoptotic human bone marrow (BM) B cells, neutrophils, Jurkat T cells, and testicular cells could readily be identified with MC540. The latter is particularly noteworthy since some of the standard methods for identifying cell death have not worked well with human cells. The versatility of this dye is such that it was also possible to phenotypically label cells stained with MC540 to analyze apoptosis in heterogenous populations of cells. Finally, the rate of detection of apoptotic cells after treatment of thymocytes with dexamethasone at 2, 4, 6, and 8 h with MC540 was shown to be equivalent to PI and annexin-V. Taken together, the data demonstrate that when proper precautions are taken, MC540 is a reliable, versatile, and inexpensive fluorochrome that can be used to identify apoptotic cells of human or murine origin even in heterogenous populations that require multicolor labeling. PMID- 11861072 TI - A competitive immunoassay to detect a hapten using an enzyme-labelled peptide mimotope as tracer. AB - Mimotope peptides-peptides which mimic the binding of a hapten to its corresponding monoclonal antibody-were conjugated to peroxidase and used in competitive immunoassay. The established immunoassay was used to quantitatively determine the concentration of hapten. As model system in all the experiments described here, we used the binding of the monoclonal antibody B13-DE1 to fluorescein and the corresponding peptide mimotope. PMID- 11861073 TI - Application of IL-5 ELISPOT assays to quantification of antigen-specific T helper responses. AB - To be able to determine the frequencies of helper-type T lymphocyte (Th) precursors specific for viral or tumor-associated antigens, an ELISPOT assay for IL-5 production was developed. The assay reproducibility was first determined using fresh or cryopreserved peripheral blood mononuclear cells (PBMC) obtained from six normal donors and tested for IL-5 production after 48 h stimulation with phytohemagglutinin (PHA). Both inter-assay (within-subject CV=1.6%) and intra assay (within-subject CV=0.8%) variabilities were found to be acceptable, and the frequencies of IL-5 secreting cells were comparable in cryopreserved and fresh PBMC of the same donors. The presence and frequency of Th precursor cells to viral capsid L1 protein (viral-like particles, VLP-L1) of human papillomavirus type 16 (HPV-16) in PBMC obtained from seven non-immunized donors and two volunteers immunized with VLP-L1 were then evaluated. Using autologous dendritic cells as antigen-presenting cells (APC) for VLP-L1 in direct 48-h ELISPOT assays, the mean frequency of IL-5 secreting CD4+ T cells was found to be <1/10(5) (negative) in normal donors but was 1/2, 436 and 1/3678 in the two volunteers immunized with VLP-L1. The assay is applicable to monitoring of the frequency of antigen-specific T cells in the peripheral circulation of individuals immunized with HPV-derived antigens. To test the assay utility in the assessment of Th tumor-reactive lymphocytes, we also used it to determine the frequency of the wild-type sequence (wt) p53(22-36) peptide-specific, HLA-DR4-restricted T cells in the bulk CD4+ T cell line. This frequency was 1/33. The ELISPOT assay for IL-5 production can be reliably used to measure Th-type responses in a variety of experimental settings. PMID- 11861074 TI - Simultaneous analysis of eight human Th1/Th2 cytokines using microarrays. AB - The adaptive immune system induces T cells to change from a naive phenotype to a Th1/Th2 phenotype each of which produce characteristic types of cytokines. Knowledge of whether a specific immune response is Th1 or Th2 is a useful indicator for diseases with basis in immune function disorder. An assay that can rapidly analyze multiple cytokines indicative of these two cell types from small sample quantities can be an extremely useful research and diagnostic tool. Silanized glass slides were printed with multiple arrays of capture antibodies to detect eight different cytokines involved in the Th1/Th2 response along with control proteins for assessing assay performance. Arrays were developed by sequential addition of known antigen amounts, detector antibodies and a fluorescent detection system followed by imaging and quantification. These arrays were used to determine the specificity, sensitivity and reproducibility of the assay and the performance compared with conventional ELISA. This multiplexed assay is able to measure human Th1/Th2 cytokines in sample volumes lower than 20 microl. The assay sensitivity for the eight cytokines range from 0.3 microg/l for IL-4 to 6.4 microg/l for IL-5 which are either comparable to or higher than those reported for conventional ELISA or bead-based multiplex ELISA methods. This assay can be automated to measure expression levels of multiple Th1/Th2 cytokines simultaneously from tens to hundreds of biological samples. This assay platform is more sensitive and has a larger dynamic range as compared to a conventional ELISA in addition to significantly reducing the time and cost of assay. This platform provides a versatile system to rapidly quantify a wide variety of proteins in a multiplex format. PMID- 11861075 TI - Development of a quantitative immuno-PCR assay and its use to detect mumps specific IgG in serum. AB - Determination of the immune status of individuals to vaccine-preventable diseases requires an assay that can detect antibodies that may be present at very low levels, especially when natural or vaccine exposure may have been many years previously. Immuno-PCR (iPCR) has recently been described as an ultrasensitive method for the detection of antigens and we have adapted the method for the quantification of antibodies to mumps virus. The procedure used was similar to an indirect ELISA except that the detecting antibody (anti-human IgG) was chemically conjugated to a short capture oligonucleotide rather than an enzyme. The capture oligonucleotide was then detected by the addition of target DNA, which was designed to hybridise to the capture oligonucleotide and function as a template for real-time PCR. The quantity of target DNA detected by the PCR depended upon the level of specific antibody in the test sample. We found that the sensitivity (and specificity) of the iPCR assay did not exceed that of the conventional ELISA. The sensitivity was limited by nonspecific binding of human IgG to the solid phase. Further development of reagents and assay formats is necessary to fully exploit the potential of quantitative iPCR, so that potential improvements in the sensitivity of anti-mumps IgG detection can be realised. PMID- 11861076 TI - Combination of MHC-peptide multimer-based T cell sorting with the Immunoscope permits sensitive ex vivo quantitation and follow-up of human CD8+ T cell immune responses. AB - Identification of MHC-restricted antigens and progress in the induction and control of adaptive cytotoxic immune responses have led to renewed interest in immunotherapy as a treatment for severe pathologies such as cancer and autoimmune diseases. Reliable procedures for detecting and monitoring T cell responses induced by the treatment throughout a clinical trial are needed in order to design rational protocols with increased efficiency. We have attempted to develop such a procedure by combining T cell sorting using HLA-peptide complexes multimerized on magnetic beads together with the quantitative Immunoscope approach. Once a recruited patient has been typed for HLA and target antigens, relevant HLA--peptide multimers can be selected and used for sorting specific peripheral T cells prior to any treatment and at the peak of the expected response to treatment. Clonotypic primers specific for the TCR rearrangements of the specific T cell clones can then be designed and used for measuring the frequency of their TCR transcripts by quantitative PCR on blood samples or T cell subsets throughout the trial. In reconstruction experiments as well as in samples from one rheumatoid arthritis patient, we were readily able to detect and follow several T cell clones with a frequency as low as 10(-5) among CD8+ T cells. The main advantages of this procedure over other currently available assays are that it does not require any assumptions on the functional status of the specific T cells and it permits the monitoring of individual T cell clones whose phenotypic shift can thus be evaluated. PMID- 11861077 TI - Analysis of spontaneous mRNA cytokine production in peripheral blood. AB - The quantification of the cytokine mRNAs synthesised by peripheral blood cells should make it possible to estimate a "peripheral immune statute". However, an accurate quantification can only be performed from a fresh whole blood sample in which mRNA is protected against nuclease digestion, and where gene transcription is inhibited. As discussed in this note, this has been made possible by the use of surfactant reagents such as tetradecyltrimethylammonium oxalate. We performed RT-PCR for the quantification of IL-10 and IFN-gamma mRNAs spontaneously produced in peripheral blood. The results showed pronounced higher IFN-gamma transcript levels in whole blood compared to peripheral blood mononuclear cells (PBMC) from the same individuals, while no significant difference was observed for IL-10 mRNA. The higher amounts of IFN-gamma mRNA observed in blood can be attributed at least to mRNA degradation. Using a real time PCR technique, we indeed demonstrated that blood IFN-gamma mRNA is rapidly degraded in vitro, the t 1/2 being worth approximately one hour at room temperature. PMID- 11861079 TI - Subtractive immunization: a tool for the generation of discriminatory antibodies to proteins of similar sequence. AB - Antibodies specific for a protein of interest are invaluable tools for monitoring the protein's structure, location and activity. Due to the tendency of an immune system to mount a response toward the abundant, immunodominant epitopes in a protein mixture, difficulties are inherent in the isolation of antibodies specific for proteins that are rare or poorly immunogenic. Likewise, isolation of antibodies specific for a protein with significant sequence similarity to other proteins, such as those derived from protein engineering, may be challenging. Subtractive immunization is a technique proven to facilitate efforts to produce monoclonal antibodies specific for antigens that are present in low abundance in a protein mixture, poorly immunogenic and/or similar in sequence or structure to other proteins. This protocol provides a detailed, stepwise procedure for the isolation of antibodies specific for a protein with sequence similarity to other proteins. As an example, we describe methods established to isolate antibodies specific to a methionine-enriched variant of soybean vegetative storage protein beta (VSPbeta-Met) that shares 91.8% amino acid sequence identity to the wild type protein (VSPbeta-WT). These methods include cyclophosphamide-induced immunosuppression of mice for the wild-type protein followed by immunization with VSPbeta-Met. As a result of this procedure, mouse polyclonal antibodies that exhibited 10-fold greater reactivity with VSPbeta-Met than VSPbeta-WT in an ELISA were generated. It is anticipated that this strategy will have utility for generating antibodies specific to protein variants derived from protein engineering. PMID- 11861078 TI - Construction and characterization of affibody-Fc chimeras produced in Escherichia coli. AB - Affibody-Fc chimeras were constructed by genetic fusion between different affibody affinity proteins with prescribed specificities and an Fc fragment derived from human IgG. Using affibody ligands previously selected for binding to respiratory syncytial virus (RSV) surface protein G and Thermus aquaticus (Taq) DNA polymerase, respectively, affibody-Fc fusion proteins showing spontaneous Fc fragment-mediated homodimerization via disulfide bridges were produced in Escherichia coli and affinity purified on protein A Sepharose from bacterial periplasms at yields ranging between 1 and 6 mg/l culture. Further characterization of the chimeras using biosensor technology showed that the affibody moieties have retained high selectivities for their respective targets after fusion to the Fc fragment. Avidity effects in the target binding were observed for the affibody-Fc chimeras compared to monovalent affibody fusion proteins, indicating that both affibody moieties in the chimeras were accessible and contributed in the binding. Fusion of a head-to-tail dimeric affibody moiety to the Fc fragment resulted in tetravalent affibody constructs which showed even more pronounced avidity effects. In addition, the Fc moiety of the chimeras was demonstrated to be specifically recognized by anti-human IgG antibody enzyme conjugates. One application for this class of "artificial antibodies" was demonstrated in a western blotting experiment in which one of the anti-RSV surface protein G affibody-Fc chimeras was demonstrated to be useful for specific detection of the target protein in a complex background consisting of a total E. coli lysate. The results show that through the replacement of the Fab portion of an antibody for an alternative binding domain based on a less complicated structure, chimeric proteins compatible with bacterial production routes containing both antigen recognition domains and Fc domains can be constructed. Such "artificial antibodies" should be interesting alternatives to, for example, whole antibodies or scFv-Fc fusions as detection devices and in diagnostic or therapeutic applications. PMID- 11861080 TI - DNA extraction using modified bacterial magnetic particles in the presence of amino silane compound. AB - Magnetic particles produced by magnetic bacteria have been used to carry out magnetic separation of DNA. Separation was achieved using magnetite coated with 3 aminopropyltriethoxysilane, N-(trimethoxy-silylpropyl) isothiouronium chloride or 3-[2-(2-aminoethyl)-ethylamino]-propyltrimethoxysilane (AEEA). The DNA binding efficiency increased with the number of amino groups present on the silane compounds and was 14 fold higher than with untreated magnetite. Addition of AEEA to aqueous solutions containing coated magnetite increased efficiency due to co condensation of DNA. From 10(8) Escherichia coli cells, 7.1 microg of DNA was recovered using 100 microg of magnetite. E. coli DNA extracted with modified bacterial magnetite was suitable for restriction enzyme digestion and polymerase chain reaction (PCR). Ten replicate samples of E. coli cells were extracted using an automated magnetic robot and 9.5 microg of DNA was extracted using 100 microg of modified bacterial magnetite and possessed a 1.94 absorbance ratio (260:280 nm). PMID- 11861081 TI - Bioelectronic device consisting of cytochrome c/poly-L-aspartic acid adsorbed hetero-Langmuir-Blodgett films. AB - A bioelectronic device consisting of protein-adsorbed hetero-Langmuir-Blodgett (LB) films was investigated. Four kinds of functional molecules, cytochrome c, viologen, flavin, and ferrocene, were used as a secondary electron acceptor (A2), a first electron acceptor (A1), a sensitizer (S), and an electron donor (D), respectively. To fabricate the cytochrome c adsorbed hetero-LB film, poly-L aspartic acid was used as the bridging molecule. The hetero-LB film was fabricated by subsequently depositing ferrocene, flavin, and viologen onto the pretreated ITO glass. Cytochrome c-adsorbed hetero-LB films were prepared by the adsorption of cytochrome c onto the poly-L-aspartic acid treated-LB films by intermolecular electrostatic attraction. Finally, the MIM (metal/insulator/metal) structured molecular device was constructed by depositing aluminum onto the surface of the cytochrome c-adsorbed hetero-LB films. Hetero-LB films were analyzed by Atomic Force Microscopy (AFM), and cytochrome c adsorption onto the films confirmed. The photoswitching function was achieved and the photoinduced unidirectional flow was in accordance with the rectifying characteristics of the molecular device. The direction of energy flow was in accordance with the energy level profile across molecular films. Based on the measurement of the transient photocurrent of the molecular device efficient directional flow of photocurrent through the redox potential difference was observed. The photodiode characteristics of the proposed bio-electronic device were verified and the proposed molecular array mimicking the photosynthetic reaction center could be usefully applied as a model system for the development of the bio-molecular photodiode. PMID- 11861082 TI - A unique approach for high level expression and production of a recombinant cobra neurotoxin in Escherichia coli. AB - In this report, we describe a simple approach to produce a large quantity of a recombinant cobra neurotoxin containing four pairs of disulfide bonds. A cDNA encoding the toxin was fused, in frame, to the carboxyl termini of thioredoxin via a linker sequence encoding two amino acids, Asp and Pro. Due to the presence of thioredoxin, a soluble form of the fusion protein was expressed in a compartment, sensitive to osmotic pressure, in Escherichia coli. The fusion protein was released into the solution with low ionic strength under an osmotic shock treatment, and purified in a single step using an ion exchange chromatography column. The purified protein was treated in diluted hydrochloric acid to induce hydrolysis of the protein at the Asp-Pro linker site. Then, the recombinant neurotoxin was purified by gel filtration of the acid-treated sample. When the biological activity of the purified toxin was assayed, it was as potent as the natural toxin. Using this protocol, approximately 12 mg of pure recombinant neurotoxin can be produced from one liter of bacterial culture. More importantly, this protocol can be easily used for the production of the toxin at a larger scale with low cost. The approach outlined in this report will be suitable for the production of other recombinant proteins especially those of the 'three-finger' family. PMID- 11861083 TI - Design of high essential amino acid proteins: two design strategies for improving protease resistance of the nutritious MB-1 protein. AB - Protein design is currently used for the creation of new proteins with desirable traits. In our lab, we focus on the synthesis of proteins with high essential amino acid content having potential applications in animal nutrition. One of the limitations we face in this endeavour is achieving stable proteins despite a highly biased amino acid content. We report here the synthesis and characterisation of two mutants derived from our MB-1 designer protein. The first mutant contains a disulphide bridge designed to cross-link remote segments of the polypeptide chain. The second one is a Tyr62-Trp mutant, where position 62 is buried in the core of the protein. Both mutants were found to be largely helical as per design, and based on thermal denaturation experiments, were substantially more stable than the MB-1 parent molecule. Enhancement of conformational stability in MB-1Trp translated into an impressive improvement of its ability to resist proteolytic degradation. Furthermore, digestion experiments intended to model degradation of proteins in a cow's rumen revealed that MB-1Trp's resistance to degradation compared to that of cytochrome c. Design strategies used for these mutants are discussed with regards to their applicability in creating efficient nutritional proteins. PMID- 11861084 TI - Enhanced adjuvantic property of polymerized liposome as compared to a phospholipid liposome. AB - Liposome, although intensively researched as vaccine or drug delivery vehicle, has been of limited use due to the low and unpredictable long-term stability. In order to overcome such problems, polymerized liposome (PL) that could initiate polymerization under very mild reaction condition was examined and compared to a conventional liposome. The polymerizable lipid, 1,2-bis[12-(lipoyloxy)dodecanoyl] sn-glycero-3-phosphorylcholine (DLL), was synthesized according to the literature, and 1,2-distearoyl-sn-glycero-3-phosphorylcholine (DSPC) was used as the conventional lipid counterpart. Polymerization of liposome was as easy and convenient as just shaking in pH 7.4 buffer. The protein encapsulation efficiency of DLL was higher than that of DSPC, and its protein release rate was lower. Immunoglobulin G (IgG) activity examined after intraperitoneal injection of antigen encapsulated by either DLL or DSPC showed that ca. 2 times as much antibody was formed by DLL-encapsulated lysozyme compared with DSPC-encapsulated form. The reasons for the superior adjuvantic properties of DLL and its future application as a drug delivery system are briefly discussed. PMID- 11861085 TI - Enzymatic properties of a neutral endo-1,3(4)-beta-xylanase Xyl II from Bacillus subtilis. AB - A Bacillus sp. CCMI 966, characterised as Bacillus subtilis, has a duplication time of about 24 min. It produces at least two extracellular xylanases, Xyl I and Xyl II. The extracellular xylanase activity seems to be strongly correlated with the biomass growth profile. The Xyl II isoenzyme was purified by ammonium sulphate precipitation and anionic exchange chromatography, with a purification factor of 8.3. The molecular weight of the isoenzyme was estimated by SDS-PAGE revealing that Xyl II is a multimeric enzyme with a catalytic subunit of about 20 kDa. Under non-denaturing conditions, a molecular weight of about 340 kDa was obtained by native PAGE gel and of 20 kDa by gel filtration chromatography. The enzyme showed an optimum pH and temperature of 6.0 at 60 degrees C. Xyl II was stable at 40 degrees C for 180 min at pH 6.0. The specificity of Xyl II for different substrates was evaluated. Xyl II presents a higher affinity towards OSX, with a K(m) of 1.56 g l(-1) and showed the ability to hydrolyse laminarin, with a K(m) of 1.02 g l(-1). Xylotetraose is the main product of xylan degradation. The Xyl II ability for binding to cellulose and/or xylan was also studied. PMID- 11861086 TI - Expression of human plasminogen kringle 5 as fusion protein with truncated hIFNgamma gene in Escherichia coli. AB - The human interferon gamma (hIFNgamma) gene was used as a fusion partner to mediate the expression of heterologous proteins and the effect of the fusion partner length on the expression of the heterologous protein was researched. Plasminogen kringle 5 (pk5), an inhibitor of angiogenesis, was fused to hIFNgamma and its serially truncated fragments, respectively, and the expression of fusion proteins was determined by SDS-Page gel. The pk5 protein was obtained readily by the introduction of sequences recognized by protease factor Xa at the fusion site and ion-exchange chromatography was employed to purify pk5. The recovery of the biological activities of pk5 was studied using the orthogonal experimental design L9 (3(4)) (four factors, three levels, nine experiments) and evaluated by measurement of anti-endothelial cell proliferation in vitro. PMID- 11861087 TI - Phage antibody fragments library combining a single human light chain variable region with immune mouse heavy chain variable regions. AB - We describe the construction of a phage antibody fragments library which combines, in a single cloning step, a synthetic human light chain variable region (V(L)) with a diverse set of heavy chain variable regions, from a mouse immunized with the prostate specific antigen (PSA). Despite V(L) restriction, selection from this library rendered two different single chain Fv antibody fragments, specifically recognizing PSA. The human V(L), used as a general partner for mouse heavy chains, was constructed by linking the germline A27 gene and the J(K)1 minigene segment, both of which are prominently involved in human antibody responses. Our approach offers a fast and simple way to produce half-human molecules, while keeping the advantage of immunizing animals for high affinity antibodies. PMID- 11861090 TI - The physics of the oscillating bubble made simple. AB - The physics of bubbles and of their oscillations is extremely complex and attempts at its mathematical description are generally inaccessible. Yet some idea, in broad descriptive terms at least, is very helpful in understanding bubble phenomena of clinical interest. A brief attempt to provide such a description is given in this article. PMID- 11861088 TI - Single-cell viability assessment with a novel spectro-imaging system. AB - Single-cell viability assessment by means of plural dye probes require the spectral and temporal analysis of microscopic images of the test cells. To meet this requirement, we have developed a simple and compact spectro-imaging system using an image slicer and a grism. The image slicer was made of a bundle of 100 optical fibers. The field of view is divided into 10 x 10 sections. The spectral data of each section could be recorded every 5 s in the range from 400 to 800 nm at 5 nm resolution. The viability changes of yeast or tobacco single-cells were measured with this system. Using BY-2 cells, for example, the response to a chemical stress of saponin was measured by means of two fluorescent probes. The spectral-spatial-temporal data of fluorescein and DNA bound ethidium bromide provided us with useful information about the dynamic change of cell membrane permeability from which the cell viability was assessed. PMID- 11861091 TI - Enhancement characteristics of the microbubble agent Levovist: reproducibility and interaction with aspirin. AB - We investigated the reproducibility of Doppler enhancement indices following intravenous bolus injections of Levovist (Schering AG, Berlin) microbubbles. We also aimed to determine whether observations from animal studies suggesting that aspirin potentiates microbubble enhancement were reproducible in humans. In five healthy volunteers, time enhancement profiles of Doppler intensity following repeated bolus injections of Levovist were acquired from the common carotid artery, hepatic vein and kidney using spectral and power Doppler before and after oral aspirin (600 mg). Peak enhancement (PE), area under the curve (AUC) and decay slopes (lambda) were calculated. Hepatic vein contrast arrival time (AT) was determined subjectively. Well-defined carotid enhancement was seen in 19/20 injections. Reproducibility was high (r > 0.8). PE and AUG were unaffected by aspirin, but lambda was slightly reduced (P = 0.02). Renal power Doppler profiles were well defined (10/10) with no significant changes of AUC, PE or lambda after aspirin. Our study demonstrates good reproducibility of carotid spectral Doppler time intensitometry with Levovist in man. Aspirin does not have a significant effect on enhancement indices except carotid spectral Doppler decay. We conclude that aspirin is unlikely to potentiate microbubble enhancement, as seen in animal studies. PMID- 11861092 TI - Which continuous US scanning mode is optimal for the detection of vascularity in liver lesions when enhanced with a second generation contrast agent? AB - OBJECTIVES: Microbubble echo-enhancers help in the assessment of focal liver masses by enhancing the signal from blood vessels. A variety of linear and nonlinear scanning modes are now available, but it is unclear which is optimal. A controlled comparison was performed during the infusion of such an agent (SonoVue: Bracco, Milan, Italy). METHODS AND MATERIALS: Ten patients with known focal liver lesions were studied. The diagnoses, confirmed on dual phase helical computed tomography (CT) at the same attendance were metastasis (n = 7), haemangioma (n = 2) and focal nodular hyperplasia FNH (n = 1). A dose of 12 ml SonoVue concentrated at 5 mg/ml was infused intravenously at a rate of 1 ml/min. The enhancement level was monitored with a continuous wave (CW) Doppler probe over the right radial artery and the intensity of the signal was registered at 1 s intervals. When a plateau of enhancement was reached, a single lesion in each patient was imaged using five different continuous scanning modes, fundamental grey scale (FGS); fundamental colour Doppler (FCD); fundamental power Doppler (FPD); second harmonic grey scale (HGS); and pulse inversion mode (Pim) using an HDI5000 scanner and C5-2 probe (ATL, Bothell, WA). The order of scanning modes was varied between patients using a predefined randomisation protocol. The videos (super video home system (SVHS)) were analysed offsite by two blinded readers, both experienced in contrast ultrasound of the liver. The readers were asked to score each mode in terms of its ability to detect vessels within/around the lesion at optimal enhancement. This was done using a ranking system (1, worst; 5, best) for each patient. RESULTS: Both observers scored FPD as the optimal imaging method, followed by Pim. (Scores summed across all patients, observer 1: FPD 48, Pim 42, FCD 37, HGS 21, FGS 10; observer 2: FPD 49, Pim 40, FCD 38, HGS 21, FGS 10). The differences from FPD were significant for FCD, HGS and FGS using a unpaired analysis of variance (ANOVA) comparison, with Bonferroni multiple corrections, (P<0.01, both observers). The differences between FPD and Pim were also significant both for observer 2 and for both observers combined (P<0.01), but did not reach significance for observer 1 (P = 0.19). CONCLUSIONS: In this study, FPD performed best, and the non-linear modes, performed continuously (pulse inversion and second HGS), showed no clear advantage. PMID- 11861093 TI - Initial observations on the effect of irradiation on the liver-specific uptake of Levovist. AB - The aim of this pilot study was to see if the biodistribution of the microbubble Levovist (SHU 508 A; Schering AG, Berlin) during its liver specific phase is altered by radiotherapy. The mechanism of this liver-specific phase of this agent remains poorly understood. One way of investigating this is to see what effect radiotherapy has on liver uptake, as both Kupffer cell function and vascular endothelial integrity are selectively damaged by irradiation. The regional liver specific uptake of Levovist was evaluated in eight patients undergoing radiotherapy to the hepatic area. Ultrasound (US) sweeps were made 4 min after Levovist injection using the phase inversion mode (PIM) which is specific for microbubbles. Differences between irradiated and non-irradiated areas were observed in 2/8 subjects completing the study. Both subjective and objective evaluations in these subjects showed a significantly reduced grey scale unit in non-irradiated versus irradiated liver regions (average values 99 vs. 89, P < 0.0045 and 75 vs. 62, P < 0.0001). These findings are somewhat inconclusive, but given the difficulty in defining areas of irradiated and non-irradiated liver, because multiple radiotherapy portals were used in all patients, tentatively suggests a radiotherapy induced effect in at least some patients. The two likely mechanisms would be damage to the Kupffer cells and or the vascular endothelium, although the relative contribution of these is unclear. PMID- 11861095 TI - Seeking consensus: contrast ultrasound in radiology. AB - This paper summarises the discussions from a meeting held on contrast ultrasound held on 21 October 2000 in Toronto, Canada. The aims of this meeting, supported by ATL/Philips Ultrasound, was to review the current clinical indications for contrast usage in the liver and kidney, arrive at recommendations for use of intravenous contrast agents, and speculate on the future uses. This paper is published to help understand this rapidly evolving field. Consensus points included a recommendation that Levovist in its post-vascular phase was of considerable value in detecting and excluding metastases in the liver, although unlikely realistically to replace CT or MR. Newer agents such as Sonovue, Definity and Sonazoid, suitable for low mechanical index (MI) imaging were also of great value and may have a further role for HCC detection. Equipment manufacturers should strive to keep improving low mechanical index modes for these agents. Promising applications for characterisation included further evaluation of lesions discovered on ultrasound and as a problem solver for CT or MR. To date no contrast agents have received approval from the FDA for radiological applications in the United States. The case for reimbursement for contrast agents was strongly supported by the panel. PMID- 11861094 TI - Multi-centre clinical study evaluating the efficacy of SonoVue (BR1), a new ultrasound contrast agent in Doppler investigation of focal hepatic lesions. AB - OBJECTIVES: SonoVue is a new ultrasound contrast agent, which consists of stabilised microbubbles of a sulphur hexafluoride gas. The aim of the study was to assess its efficacy in the Doppler investigation of focal hepatic lesions. MATERIALS AND METHODS: Seventy patients with focal liver tumours were studied. Four doses (0.3, 0.6, 1.2 and 2.4 ml) of SonoVue were administered intravenously with at least 10 min delay between each injection. A complete colour/power and spectral Doppler imaging investigation of the lesions was performed at baseline pre-dosing and after each SonoVue injection. All examinations were recorded on SVHS videotapes. Baseline and post contrast videotapes were reviewed by the on site (un-blinded) investigators and by two off-site blinded readers (a) to grade the global quality of the Doppler scans of the focal lesions vascularity and the normal parenchymal vessels (b) to measure the duration of clinically useful Doppler signal enhancement and (c) to determine the diagnostic accuracy and performance of the enhanced versus unenhanced scans using histopathology, tumour markers, CT and/or MR as the reference standard. RESULTS: A statistically significant improvement was observed at all four SonoVue doses in the off site assessment of global quality of the Doppler examination of tumoral and normal parenchymal vessels in comparison with the baseline (P < 0.05). The median duration of clinically useful enhancement was significantly increased with increasing doses (P < 0.001), ranging between 1.4-2.2 min for the lowest dose and 3.2-3.8 min for the highest dose for the off-site readers. On-site assessment of diagnostic accuracy showed a significant increase in the specificity of the Doppler diagnoses (P < 0.0016) with an increase in the positive and negative predictive values and in the likelihood ratio in differentiating between benign and malignant lesions. Off-site evaluation showed a significant increase in the accuracy of enhanced Doppler diagnosis in comparison with the baseline performance. CONCLUSION: The results suggest that SonoVue is effective in improving the display of tumoral vascularisation and may be useful in the characterisation of focal liver lesions. PMID- 11861096 TI - Ultrasonic contrast agents: safety considerations reviewed. AB - Ultrasonic contrast agents are usually comprised of a stabilised shell encapsulating a gas bubble. When these are introduced in the body they increase the acoustic scattering from the tissues through which they pass, and especially from the vasculature. Their primary uses lie in cardiological and oncological imaging. However, these microbubbles have the potential to act as centres for acoustic cavitation activity, and so it is important to consider the safety of their use from an acoustic standpoint. The addition of ultrasonic contrast agents to in vitro suspensions of red blood cells has been shown to lead to haemolysis when the sample is exposed to ultrasound at levels which leave the cells unharmed in their absence. In vivo the infusion of gas bubble contrast agents into experimental animals has been shown to enhance the incidence of petechiae and haemorrhage in the intestine. The Mechanical Index (MI) thresholds for the effects seen in vitro lie within the range of MIs available with diagnostic clinical scanners, but in vivo the thresholds lie at the top end of the exposure levels available clinically. No adverse effects in humans arising from the ultrasonic exposure of these contrast agents have been reported to date. PMID- 11861097 TI - The value of mathematical modelling in understanding contrast enhancement in CT with particular reference to the detection of hypovascular liver metastases. AB - OBJECTIVE: Few subjects in body imaging have generated such intensive debate as that surrounding the optimising of strategies for the detection of liver metastases using contrast-enhanced Computed Tomography (CT). Despite this, research on the subject has been almost entirely focused on experimental studies, little attention having been paid to the value of theoretical analyses. The authors' aim was to develop a relatively simple and robust mathematical model which could be implemented on a personal computer. METHODS: Simple differential equations describing the distribution of contrast agent between intra- and extra vascular spaces during an infusion are set up. An analytic solution is obtained for the plasma/blood concentrations as a fraction of time and a solution for the interstitial concentrations as a fraction of time is obtained by converting the differential equations to difference equations which are solved in a stepwise manner. RESULTS: Vascular and hepatic parenchymal enhancement-time curves are generated which are in close agreement with expectations. The model may be used to compare different infusion regimes, e.g. lower versus higher dose, faster versus slower infusions, monophasic versus biphasic infusions and scan start delay. The implications of the results of the model for clinical protocol design are discussed and the special value of spiral/helical technology indicated. CONCLUSION: A simple mathematical model has been developed to model blood and tissue contrast enhancement in CT during contrast infusion. The model clarifies a number of issues related to contrast enhancement regimes for the study of the liver and these have been discussed. PMID- 11861098 TI - A case with primary amyloidosis of the liver and spleen: radiologic findings. AB - A case with primary amyloidosis of the liver and spleen is presented. In the liver, ultrasonography showed heterogeneous echogenicity. Portal phase CT showed heterogeneous enhancement, and delayed phase CT showed delayed enhancement with focal hypoattenuating areas. Bone scintigraphy showed diffuse accumulation. In the spleen, arterial phase CT showed the lack of parenchymal enhancement, and T2 weighted MR imaging showed decreased signal intensity. The combination of imaging findings of the liver and spleen was suggestive of amyloidosis. PMID- 11861099 TI - Medial meniscocapsular separation: MR imaging criteria and diagnostic pitfalls. AB - Various magnetic resonance imaging (MRI) signs can be used in the diagnosis of medial meniscocapsular separation. A thorough knowledge of the normal anatomy of the medial meniscocapsular junction is essential to understand these signs. MRI signs used in the diagnosis of meniscocapsular separation include meniscal displacement relative to the tibia, meniscal corner tear, perimeniscal fluid, irregular meniscal outline, meniscofemoral and meniscotibial extension tears, and interposition of contrast medium between the meniscus and the MCL. Potential causes of false positive diagnosis of meniscocapsular separation include MCL bursitis, meniscal cyst, MCL tear, joint effusion, and perimeniscal fat. PMID- 11861103 TI - Isoflavone content of the soy based supplements. AB - A large number of soy isoflavone products with indications of possible health effects are available on the market. Fifteen different soy based products were analyzed using high performance liquid chromatography (HPLC) with coulometric electrode array detector to determine the total amount of isoflavones in aglycones after the hydrolysis and identify the different forms of the isoflavone conjugates. The aim of the study was to evaluate how well the isoflavone content data supplied by the producers correspond to our analysis results. Only one product contained isoflavones measured in aglycones the same amount as was the value given by the producer. The total amount of the isoflavones in aglycones ranged from 0.121 to 201 mg/g. Measured amounts of isoflavones in aglycones after the hydrolysis were in general lower than the values in the product labels. Product data were often confusing and the concrete amount of isoflavones was difficult to find out. PMID- 11861105 TI - Determination of tacrine metabolites in microsomal incubate by high performance liquid chromatography-nuclear magnetic resonance/mass spectrometry with a column trapping system. AB - A column trapping system has been incorporated into high performance liquid chromatography-nuclear magnetic resonance-mass spectrometry (HPLC-NMR-MS) to reduce data acquisition time of NMR experiments. The system uses a trapping column to capture analytes after the HPLC column and back flush trapped analyte to the flow cell of the NMR probe for detection. A dilution solvent is mixed with eluent from HPLC column to reduce the influence of the organic content in the mobile phase before column trapping. The trapping column is also coupled with a mass spectrometer (MS) to get complementary MS data on the same peak. Studies on 1-hydroxylated 9-amino-1,2,3,4-tetrahydro-acridine (1-OH tacrine), indomethacin and testosterone with the column trapping system showed good recovery of analytes and over 3-fold mean increase in UV-VIS signal intensity. The time saving on NMR experiments with the column trapping system was demonstrated by the analysis of dog microsomal incubate with tacrine. PMID- 11861104 TI - Preparation of beta-artemether liposomes, their HPLC-UV evaluation and relevance for clearing recrudescent parasitaemia in Plasmodium chabaudi malaria-infected mice. AB - Egg phosphatidylcholine-cholesterol liposome formulations containing the antimalarial drug beta-artemether have been prepared and analyzed for their encapsulating capacity, chemical stability, leakage, in vitro release and their therapeutic efficiency against Plasmodium chabaudi infection. A HPLC-UV analysis of beta-artemether liposomes without derivatisation was achieved. A good linearity of y=4437.7 x+469.01 (R(2)=0.9999) with a detection limit of 2 microg ml(-1) was reached. Prior to this, liposomal formulations composed of different molar ratios of EPC-CHOL were prepared to select beta-artemether crystal-free liposome preparations. The formulation corresponding to 4:3 and a total concentration of 300 mg lipids ml(-1) buffer (pH 7.2), which could incorporate as much as 1.5 mg beta-artemether was selected for therapy. A trapping efficiency of nearly 100% was reached, the drug being located in the lipid bilayers. A dialysis test demonstrated that the drug could be reversibly released from the liposomes, reaching equilibrium within 24 h. After 3 months storage at 4 degrees C, no leakage of beta-artemether had occurred indicating a high stability of the liposomes. These liposomes were used to treat mice infected with the virulent rodent malaria parasite Plasmodium chabaudi chabaudi, with a 100% cure by clearing the recrudescent parasitaemia. PMID- 11861106 TI - A comparison of quantitative NMR and radiolabelling studies of the metabolism and excretion of Statil (3-(4-bromo-2-fluorobenzyl)-4-oxo-3H-phthalazin-1-ylacetic acid) in the rat. AB - The identification and quantitation of the metabolites of Statil in rat bile and urine were investigated by 1H- and 19F-NMR spectroscopy and liquid scintillation counting. Male Wistar rats received a single oral dose of 100 mg/kg of radiolabelled Statil. Statil is known to produce glucuronide conjugates which are predominantly excreted into the bile in male rats. The complex multiphasic matrix of bile has been shown to make identification of the resonances by 1H-NMR spectroscopy very difficult as Statil appeared to be micellar bound giving rise to very broad signals. This not only impaired unambiguous signal characterisation but also quantification. The partial separation by SPEC-(1)H-NMR spectroscopy enabled the disruption of the micellar matrices and hence enabled the identification of Statil predominantly as aglycone, and to a lesser extent as glucuronide conjugate. In addition, minor acyl migration products of Statil glucuronide could also be detected as they were separated during the SPEC process. 19F-NMR spectroscopic measurements on whole bile confirmed their presence as a number of overlapped signals could be observed. The selectivity, simplicity and signal dispersion characteristic of 19F-NMR spectroscopy also enabled the calculation of dose related recoveries of Statil related material in the bile and urine samples without the need for a radiolabel. The aim of this work was to investigate the usefulness and limitations of NMR spectroscopy of intact bile and urine as a means of quantifying levels of drug metabolites. The results obtained from NMR spectroscopy are compared with those obtained using scintillation techniques. Scintillation counting yields unequivocal quantification results, provided the label is preserved in metabolites as has been the case here. In general, quantification by 19F-NMR results similar to those obtained by scintillation counting (in agreement within about 20%). However, discrepancies have been observed with very small and broad 19F-NMR signals in bile. Nevertheless, 19F-NMR spectroscopy of bile is a rapid and facile method for assessing metabolite levels of fluorinated drugs. PMID- 11861107 TI - Investigation of critical factors for the resolution of SR695, a key impurity, from efavirenz in the reversed-phase assay of efavirenz dosage forms. AB - An investigation of the critical factors effecting the resolution of SR695 from efavirenz in the assay of efavirenz by reversed-phase HPLC was performed. This study was implemented to address the inability of a subset of the Zorbax SB-CN columns used in this method to adequately perform this separation, which were otherwise indistinguishable from columns of this type that could. In this study, column temperature, detector time-constant, pre-gradient isocratic hold-time, pre column mixing volume, column, and HPLC type were considered. Experimental Design methods were employed to find the relative importance of these factors and to find parameters that would optimize the resolution of SR695 and efavirenz on any HPLC, with any column of this type, for both efavirenz oral liquid and capsule samples. It was also desired that this method change be minimal, so that extensive revalidation would not be required. The most important factors were the column temperature, with lower temperatures giving better resolution, and pre column mixing volume of sample with mobile phase, with higher mixing volumes giving better resolution up to an asymptote reached at around 150 microl. Added pre-gradient isocratic hold time was found to result in a small improvement in resolution, but was insignificant compared with the other factors mentioned above. A possible explanation is given for the mechanism by which temperature and pre-column mixing have this effect on the resolution obtained in this assay. PMID- 11861108 TI - Benzophenone-3: rapid prediction and evaluation using non-invasive methods of in vivo human penetration. AB - The study described in this paper constitutes a practical assay system to evaluate in vivo drug penetration using two complementary non-invasive methods. An electrical capacitance test was first applied to the skin on the forearm to evaluate the hydration of the skin, and check the integrity of the stratum corneum. In the first step, the percentage absorption was measured using an occlusive and difference method; following benzophenone-3 application any residual formulation was washed off and the amount removed analyzed. In the second step, the tape stripping method-a useful procedure for selectively removing the skin's outermost layer, the stratum corneum, and measuring the stratum corneum adsorption-was performed. Under these conditions the human skin permeation of this UV-filter over four hours was near to 35% of the applied dose with the occlusive method. The amount of topically applied benzophenone-3 found in the stratum corneum after 30 min exposure using the stripping procedure was evaluated at 4% to the applied dose. PMID- 11861109 TI - Pharmacokinetic comparison of intravenous carbendazim and remote loaded carbendazim liposomes in nude mice. AB - Carbendazim is a novel anticancer agent. The aim of this study was to prepare and characterize a remote loaded liposome preparation of carbendazim, and compare its pharmacokinetic profile to that of unencapsulated carbendazim. Carbendazim was encapsulated in liposomes composed of sphingomyelin-cholesterol (3:1, w/w) by remote loading in response to a transmembrane pH gradient (pH 0.5 in/pH 4.0 out), which resulted in encapsulation of more than 95% of the available drug in preformed vesicles. High drug/lipid ratios were prepared which correspond to a molar ratio of up to 0.8. Physical isolation of the free drug and dialysis were used to determine the in vitro release of carbendazim from liposomes. The release was independent of the initial drug/lipid ratio and choice of internal buffer composition. Liposomal carbendazim (200 mg kg(-1)) was intravenously administered to athymic nude mice and the serum levels of free carbendazim were determined by HPLC analysis after a methanol-induced protein precipitation. Administration of liposomal carbendazim to mice resulted in significant alterations in the pharmacokinetics. Serum levels of free carbendazim were approximately 10-fold greater than those achieved for the same dose of unencapsulated drug. Liposomal carbendazim showed both high C(max), AUC and low clearance rate. Liposomal carbendazim and unencapsulated carbendazim displayed a similar terminal half-life (43-48 min). The relatively large volume of distribution of carbendazim suggests that the compound may partially enter cells or be bound to some extravascular structures. The results indicate that the liposomal formulation of carbendazim significantly increases its blood concentrations. PMID- 11861110 TI - High-throughput miniaturized immunoassay for human interleukin-13 secreted from NK3.3 cells using homogenous time-resolved fluorescence. AB - A miniaturized immunoassay for human interleukin-13 (IL-13) using homogeneous time-resolved fluorescence (HTRF) has been developed. In this assay, IL-13 which was secreted from NK3.3 cells stimulated with interleukin-2 (IL-2) was detected by measuring the time-resolved fluorescence after adding a mixture of three reagents, biotinylated anti-IL-13 monoclonal antibody, europium cryptate (fluorescence donor)-labeled different anti-IL-13 monoclonal antibody and crosslinked allophycocyanin (fluorescence acceptor)-conjugated with streptavidin in a 384-well assay plate. The detection limit of IL-13 using this immunoassay was estimated to be less than 600 pg/ml and IL-13 levels measured by this method were very close to those measured by enzyme linked immunosorbent assay (ELISA; the correlation coefficient was 0.9535). The proposed assay requires only a fourth of the quantities of all reagents compared with the assay using a conventional 96-well microtiter plate. Furthermore, there is no need to transfer the culture supernatant to another assay plate and wash the plate. Therefore, this miniaturized immunoassay is economical and efficient and is particularly suitable for high-throughput drug screening. PMID- 11861111 TI - Determination of iodine values according to Hanus using 1,3-dibromo-5,5 dimethylhydantoin (DBH): analytical methods of pharmacopeias with DBH: part 7. AB - USP/NF 2000 [The United States Pharmacopeia, Rockville USA, 24th ed., 2000, p. 1868, The National Formulary, 19th ed. 2000] and PH. EUR. 1997 [European Pharmacopoeia, third ed., Council of Europe, Strasbourg, 1997, pp. 63-64] determine the iodine values according to Hanus with iodine monobromide in glacial acetic acid. This reagent can be replaced by a solution of 1,3-dibromo-5,5 dimethylhydantoin (DBH) and potassium iodide or iodine in the same solvent. Both reagents yield equivalent results by means of method comparison according to Passing and Bablok [J. Clin. Chem. Clin. Biochem. 21 (1983) 709; 22, (1984) 431] in relation to the official method of PH. EUR. PMID- 11861112 TI - Application of 1D 1H NMR for fast non-targeted screening and compositional analysis of steroid cocktails and veterinary drug formulations administered to livestock. AB - A fast non-targeted strategy is described for analysis of formulations--meant for administration to live stock--containing growth-promoting agents or veterinary drugs. The use of 1H NMR as a first step universal screening method is applied and used in routine analysis. The implementation of this approach has increased the analysis efficiency considerably. Apart from screening on illegal compounds, 1H NMR information on matrix and thus, indirectly, administration mode, can be present. An ever-growing 1H NMR database is used containing more than 200 reference substances. Based on the 1H NMR screening, decisions for further analysis can be made, such as for instance HPLC fractionation of steroid cocktails and subsequent 1H NMR (and LC-MS) analysis. Examples of unravelling formulations are given in detail including a steroid cocktail containing 15 compounds. PMID- 11861113 TI - Direct determination of pindolol enantiomers in human serum by column-switching LC-MS/MS using a phenylcarbamate-beta-cyclodextrin chiral column. AB - A direct analytical method of pindolol enantiomers in body fluids was developed by means of column-switching semi-microcolumn liquid chromatography/tandem mass spectrometry (LC-MS/MS). A pre-column packed with a silica-based cation-exchanger was used for on-line sample cleanup. Subsequent enantioseparation was conducted with a phenylcarbamate-beta-cyclodextrin (ph-beta-CD) bonded semi-micro chiral column (2.0 mm inner diameter (i.d.)). A 25-microl aliquot of serum/urine samples was directly injected into the system after simple filtration with a membrane filter. Separated enantiomers were monitored with positive electrospray ionization (ESI) and selected reaction monitoring (SRM). R(+)- and S(-)-pindolol in serum and urine were determined separately within 16 min at a resolution factor of 1.9. The detection limits at a signal-to-noise (S/N) ratio of 5 were 0.13 ng ml(-1) for both enantiomers. The linearity of the method was in the range of 0.25-100 ng ml(-1) with regression coefficient greater than 0.997. Recoveries from spiked serum and urine samples, estimated by the external standard method, were between 94.8 and 117.6% with a relative standard deviation (RSD) ranging from 2.1 to 18%. PMID- 11861114 TI - Chiral CE separation of warfarin in albumin containing samples. AB - A capillary zone electrophoresis method with ultraviolet (UV) absorbance detection for the chiral separation of warfarin enantiomers using highly sulfated beta-cyclodextrin (beta-CD) was developed and optimized. Enantiomeric separation of warfarin was characterized by high resolution and efficiency. The optimized electrophoretic conditions were subsequently applied to the analysis of warfarin extracted from spiked human serum albumin samples. This assay showed acceptable precision, with linearity in the warfarin enantiomer concentration range of 0.1 25 mg/l. The limits of detection (LOD) and quantitation (LOQ) evaluated as warfarin enantiomer concentrations in the serum samples were 0.05 and 0.15 mg/l, respectively, for each warfarin enantiomer. PMID- 11861115 TI - Structure elucidation and conformational properties of eprosartan, a non peptide angiotensin II AT1 antagonist. AB - A novel approach to treat hypertension is to interfere with the Renin-Angiotensin system (RAS) by blocking the binding of vasoconstrictive hormone Angiotensin II to the AT(1) receptor site. This approach led to the beneficial drug losartan (COZAAR) and other similar in structure to the antihypertensive drugs (sartans). In an effort to compare the stereoelectronic features of pharmacophoric segments of the different sartans, a research activity was initiated in our laboratory related to the conformational properties of these drugs. In a previous study, the structural features which determine the pharmacophoric segments of losartan were examined. In this study, the conformational properties of eprosartan (TEVETEN), a drug with fewer side effects, were examined. In addition, the superimposition ability of losartan and eprosartan with the peptide antagonist sarmesin was studied. PMID- 11861116 TI - Employment of on-line FT-IR spectroscopy to monitor the deprotection of a 9 fluorenylmethyl protected carboxylic acid peptide conjugate of doxorubicin. AB - A method for accurately determining the end-point, >98% conversion, of the deprotection reaction of a highly toxic 9-fluorenylmethyl (Fm) ester 1b to its corresponding carboxylate 1d in real time by FT-IR spectroscopy is reported. Advantages of this method over analysis by conventional chromatographic means include real time determination of the end-point of a reaction that is time sensitive to by-product formation, and elimination of sampling a highly toxic reaction mixture. The FT-IR method is based on monitoring, in real time, the disappearance of the Fm ester carbonyl band for 1b at 1737 cm(-1), during deprotection by piperidine, and calibration models were established by Partial Least Squares (PLS) regression analysis with high performance liquid chromatography (HPLC) as reference. The best calibration model was built with 5 PLS factors in the spectral range of 1780-1730 and 1551-1441 cm(-1) and resulted in a standard error of cross validation (SECV) of 0.63 mM 1b and a standard error of prediction (SEP) of 0.51 mM 1b in the range of 0-25 mM. This error of prediction is approximately 0.8% of the initial concentration of 1b and is well within our specifications of <2% initial concentration. PMID- 11861117 TI - Moisture assay of an antifungal by near-infrared diffuse reflectance spectroscopy. AB - Near-infrared (NIR) diffuse reflectance spectroscopy was employed in the method development and validation of a moisture assay for the novel antifungal caspofungin acetate. Spectra were obtained over the entire spectral region available (950-1650 nm) using an InGaAs photodiode array detector equipped with a diffuse reflectance probe. No sample pre-treatment was required and the analysis time was less than 1 min. Primary reference data were obtained using a Karl Fischer (KF) titration (coulometric, volumetric or both). The investigated range of water content was 2.6-9.9% (w/w) with a standard error of prediction (SEP) of 0.2%. The predictive capabilities of the partial least-squares (PLS) regression calibration model used in the moisture assay were verified using independent test sets. The NIR predicted values of the developed method were equivalent to the reference method sets and the prediction error was equivalent to the reference method error. These results reveal that the predictive model constructed by means of a PLS regression is valid, rugged and could be used to determine moisture levels on-line in caspofungin acetate drug substance. PMID- 11861118 TI - Method development and validation for the GC-FID assay of tributyl phosphate in a phospholipid emulsion. AB - This paper describes the development and validation of an isothermal GC-FID method for the assay of tributyl phosphate in a phospholipid emulsion. The emulsion is used as a topical ointment to deliver Triton X-100, a spermicide. The tributyl phosphate is added to the emulsion as a plasticizer or softening agent. The chromatographic conditions of the method employ a J&W DB-Wax capillary column (30 m x 0.53 mm, film thickness 1 microm), isothermal elution with He at a column flow of 2.0 ml/min, injector, detector, and oven temperatures at 210 degrees C, a split ratio of 18.0/2.0, and a 3-microl injection volume. Sample calibration was performed with tributyl phosphate purchased from Aldrich (USP Reference Standard is not available). The linearity of the tributyl phosphate peak area responses was demonstrated from approximately 50 to 150% of the analytical concentration of 100 microg/ml. System precision was determined from five replicate injections of a standard and sample solution. Reproducibility of the tributyl phosphate peak area responses showed R.S.D. of 1.2 and 0.4%, respectively. Method precision was performed by assaying five samples by two different analysts on different days. The mean %LC was 95.5% (R.S.D.=1.0%) for the first analyst, and 95.6% (R.S.D.=1.0%) for the second analyst. The mean %LC value for all ten sample preparations was 95.5% (R.S.D.=0.9%). The limits of detection and quantitation were determined to be 0.2 and 0.7 microg/ml, respectively. PMID- 11861119 TI - Spectrophotometric methods for the rapid determination of menadione and menadione sodium bisulphite and their application in pharmaceutical preparations. AB - Two simple, rapid and sensitive spectrophotometric determination of menadione and its sodium bisulphite derivative (MSB) have been carried out. The first method involves the reaction of menadione and its sodium bisulphite derivative with 3 methyl-2-benzothiazolinone hydrazone hydrochloride (MBTH) is sodium hydroxide medium to give blue coloured product having maximum absorption at 625 nm and the coloured species is stable for more than 1 h. The Beer's law is obeyed in the range 0.4-16 microg ml(-1). The second method proposes the reaction of menadione and its sodium bisulphite derivative with resorcinol in concentrated sulphuric acid medium to give red coloured product having maximum absorption at 520 nm and is stable for 3 h. The Beer's law was obeyed in the range of 1-24 microg ml(-1). Molar absorptivity for the above two methods were found to be 7.6 x 10(3) and 4.5 x 10(3) l mol(-1) cm(-1), respectively. All the measurements were carried out at room temperature. These two methods have been successfully applied for menadione and its sodium bisulphite derivatives in injections and tablets of pharmaceutical formulations. The results compare favourably with official method. PMID- 11861120 TI - Rapid determination of acetazolamide in human plasma. PMID- 11861121 TI - Spectrophotometric determination of enrofloxacin and pefloxacin through ion-pair complex formation. AB - Two simple, quick and sensitive spectrophotometric methods are described for the determination of enrofloxacin and Pefloxacin. The methods are based on the reaction of these drugs with bromophenol blue (BPB) and methyl orange (MO) in buffered aqueous solution at pH 2.3-2.5 in case of bromophenol blue and at pH 3.6 with MO to give highly coloured complex species, extractable with chloroform. The coloured products are quantitated spectrophotometrically at 420 and 424 nm for BPB and MO, respectively. Optimisation of the different experimental conditions is described. Beer's law is obeyed in the concentration ranges 2-12 and 2-18 microg ml(-1) with BPB and in the ranges 1-12 and 4-40 microg ml(-1)with MO for enrofloxacin and pefloxacin, respectively. The proposed methods are applied for determination of Enroxil oral solution, Peflacine tablets and Peflacine ampoules with mean percentage accuracies 99.5+/-0.99, 99.39+/-1.05 and 100.02+/-0.895, respectively, with BPB and 100.30+/-0.89, 100.25+/-0.98 and 100.20+/-0.72, respectively, with MO. PMID- 11861122 TI - LC determination of praziquantel in human plasma. AB - A simple high-performance liquid chromatographic (HPLC) method for the determination of praziquantel in human plasma was developed and validated. The present method was described by adding drop-wise 0.2 M Zinc sulfate and acetonitrile to plasma sample for deproteinization. This method used a reversed phase Spherisorb ODS 2 column (5 microm), 250 x 4.6 mm i.d. as a stationary phase with a mobile phase consisting of acetonitrile- methanol-water (36:10:54, v/v/v), a flow rate of 1.5 ml/min and UV detection wavelength of 217 nm. Diazepam was used as internal standard. The standard calibration curve was linear over the concentration range of 100-2000 ng/ml (r=0.999). The equation of a linear regression line was y=8.05E-04+7.25E-04x with slope and intercept values of 0.0007 and 0.0008, respectively. The limit of detection was 12.25 ng/ml and the limit of quantification was set at 100 ng/ml. The intra- and inter-day assay coefficients of variation (CV) were 3.0+/-1.7 and 6.3+/-1.9%, respectively. The percentage of recovery was 102.1+/-5.6. Therefore, the HPLC method described here was simple, rapid and reproducible since it did not require extraction and evaporation processes in sample preparation, which will reduce time-consuming or expensive sample preparation. PMID- 11861123 TI - Stimulus-dependent activation of c-Fos in neurons and glia in the rat cerebellum. AB - The intent of the present study was to use chemical or electrical stimulation of cerebellar afferents to determine how different stimulation paradigms affect the pattern of activation of different populations of neurons in the cerebellar cortex. Specifically, we analyzed immediate changes in neuronal activity, identified neurons affected by different stimulation paradigms, and determined the time course over which neuronal activity is altered. In the present study, we used either systemic (harmaline) or electrical stimulation of the inferior cerebellar peduncle (10 and 40 Hz) to alter the firing rate of climbing and mossy fiber afferents to the rat cerebellum and an antibody made against the proto oncogene, c-fos, as a marker to identify activated neurons and glia. In control animals, only a few scattered granule cells express nuclear Fos-like immunoreactivity. Although no other cells show Fos-like immunoreactivity in their nuclei, Purkinje cells express Fos-like immunoreactivity within their somatic and dendritic cytoplasm in control animals. Within 15 min of chemical or electrical stimulation, numerous granule and glial cells express Fos-like immunoreactivity in their nuclei. Cells in the molecular layer express Fos-like immunoreactivity following harmaline stimulation in a time and lobule specific manner; they do not appear to be activated in the electrical stimulation paradigm. Following harmaline injections, there is an initial loss of Fos-like immunoreactivity in the cytoplasm of Purkinje cells; 90 min later, nuclear staining is observed in a few scattered Purkinje cells. Following electrical stimulation, the cytoplasmic staining in Purkinje cells is enhanced; it is never present in the nucleus. Data derived from this study reveal cell-specific temporal and spatial patterns of c Fos activation that is unique to each paradigm. Further, it reveals the presence of an activity dependent protein in the cytoplasm of Purkinje cell somata and dendrites. PMID- 11861124 TI - Protein synthetic machinery in the dendrites of the magnocellular neurosecretory neurons of wild-type Long-Evans and homozygous Brattleboro rats. AB - There is growing evidence of local protein synthesis in neuronal dendrites, especially in relation to synaptic activity. The hypothalamic magnocellular system is a robust model for peptidergic neurons, especially for the study of dendrites. Quantitative electron microscopy, immunocytochemistry and non radioactive in situ hybridization (with tyramide signal amplification) were used to compare dendrites of magnocellular neurons in the supraoptic nucleus of wild type rats and of homozygous Brattleboro (BB) rats which are subject to long-term hyper-osmotic stimulation because they cannot secrete vasopressin. The dendrites contained free polyribosomes, cisterns of rough endoplasmic reticulum (ER) and small Golgi-like elements. These were clustered in the dendrites, mostly near the plasma membrane. All were increased in amount in the enlarged dendrites of the BB rats. The presence of polyribosomes and cisterns of rER implies that both cytosolic and membrane-inserting proteins are synthesized in the dendrites. The ER marker protein disulfide isomerase extended far into dendrites, but Golgi element markers (mid-Golgi and trans-Golgi network) were distributed mainly in their proximal parts. In BB rats, all the labeling was stronger. 28S rRNA, initiator tRNA(Met), and poly(A) mRNA were revealed extending into proximal and middle parts of dendrites where intensely reactive punctate structures were common. 28S rRNA could be detected in the distal parts of the dendrites. The length of positively stained dendrites was increased significantly for all these RNAs in BB rats. The results provide morphological evidence that magnocellular dendrites have the capacity for local protein syntheses and that this is increased in chronic hyperosmotic stress. PMID- 11861125 TI - Expression of c-Fos, ICER, Krox-24 and JunB in the whisker-to-barrel pathway of rats: time course of induction upon whisker stimulation by tactile exploration of an enriched environment. AB - Modified tactile information has been shown to induce adaptive plasticity in the somatosensory cortex of rat. The cellular mechanisms resulting in plastic neuronal responses, however, are largely unknown. Inducible transcription factors have been proposed as one major link in the cascade from modified input to altered neuronal structure and function. We investigated the spatial and temporal patterns of transcription factor induction in the rat whisker-to-barrel pathway by placing the animals in a novel, enriched environment while having clipped sets of whiskers on one side of the face. Such stimulation resulted not only in a specific c-Fos induction in brainstem barrelettes and thalamic barreloids, but also in the barrel-related cortical columns, each with different time courses. In the barrel cortex, c-Fos and Krox-24 immunostaining showed a rapid induction with peak levels at 1 h and a return to basal levels after 14 h. JunB was induced after 1 h of exploration, declined at 6 h and returned to basal levels after this time point. The inducible cyclic AMP early repressor (ICER), a transcription factor of the cAMP signaling pathway, showed a maximum after 6 h, decreased slowly, but elevated levels were still detectable after 5 days. Our data demonstrate that upon whisker stimulation by exploration of a novel, enriched environment, (i) subcortical relay stations in the whisker-to-barrel pathway are able to express elevated levels of c-Fos and (ii) in the barrel cortex c-Fos, JunB, Krox-24 and ICER are differentially regulated in the temporal domain. PMID- 11861126 TI - Enriched environmental conditions reverse age-dependent gliosis and losses of neurofilaments and extracellular matrix components but do not alter lipofuscin accumulation in the hindlimb area of the aging rat brain. AB - We provide a description of a correlation of lipofuscin accumulation and expression of glial fibrillary acidic protein in the cerebral cortex of aged rats. Glial fibrillary acidic protein showed a complementary distribution pattern to perineuronal nets, visualized with Wisteria floribunda agglutinin. With progressing age (12-36 months), a strong increase of lipofuscin and gliosis occurred in functionally characterized cortical areas, whereas a concomitant, area-specific loss of perineuronal nets was found in the cortical somatosensory representation of the hindlimbs. In contrast to lipofuscin accumulation and increased gliosis, the loss of perineuronal nets and the reduction of non phosphorylated neurofilament H were in part reduced or prevented by housing the animals under enriched environmental conditions between 33 and 36 months of age. Especially the reduction of astrocytosis by 20% which coincided with a reduction in the loss of extracellular matrix components involved in forming the glia neuron-interface demonstrates, that the aging cortex retains its potential for functional plasticity. PMID- 11861128 TI - Effect of posterior temporal-parietal hematoma on orbital frontal chemistry in relation to a cognitive and anxiety state: a combined 1H-MRS and neuropsychological study of an unusual case as compared with 16 healthy subjects. AB - The authors report the unusual case of a 58-year-old woman (MJP) suffering from left temporal throbbing headache, associated with confusion. Magnetic resonance imaging showed a 5 x 3 x 2 cm hematoma at the left posterior temporal--parietal junction (PTPJ). Repeated MRI of MJP's brain performed during a 4-month follow-up period showed decrease in hematoma size (2.3 x 1.5 x 1) with evidence for development of encephalomalacia and resorption of blood products involving the area of hemorrhage. MJP had mild transcortical sensory aphasia characterized by difficulty with reading and processing, with semantic paraphasic errors while speaking and some difficulty with repetition. MJP had remained normotensive and seizure free, on Vasotec therapy and Dilantin prophylaxis. An in vivo proton magnetic resonance spectroscopy (1H-MRS) performed during an 8-month follow-up period showed reduced concentration for N-acetyl aspartate (NAA) by 19.3% (F=4.09, P<0.04), and myo-inositol by 32.0% (F=5.16, P<0.02) in the left orbital frontal cortex (OFC) as compared with 16 healthy subjects (age- and sex-matched). Cognitive tests (the Wechsler abbreviated scale of intelligence (WASI) and the Stroop color--word interference) showed a significant impairment suggesting involvement of higher-order cognitive functioning (memory, learning, and general intelligence) and attentional system. The Spielberger state-trait anxiety inventory (STAI) showed increased anxiety at the moment of the current examination and decreased tendency to be anxious over a long period of time. The Beck Anxiety and Depression Inventory revealed minimal anxiety and mild to moderate levels of depression. It is hypothesized that the PTPJ hematoma triggered long-distance pathways linking PTPJ area and frontal lobe, including OFC, which resulted in abnormal chemical changes in the left OFC and in cognitive tests impairment, and in long-term anxiety state changes. PMID- 11861127 TI - Reelin immunoreactivity in the larval sea lamprey brain. AB - In order to analyze the presence of a reelin-like protein in the brain of a primitive vertebrate with a laminar-type brain, such as the sea lamprey, Western blot and immunohistochemical approaches were employed by using the G10 and 142 reelin-specific monoclonal antibodies. Western blots of lamprey brain extracts showed bands of about 400 kDa, 180 kDa and others below 100 kDa; similar bands were observed in samples from rat cerebellum. In different larval stages there was a prominent reelin immunolabeling associated with the olfactory bulb, pallial regions, habenula, hypothalamus and optic tectum. In addition, the olfactory and optic tracts, as well as the afferent and efferent (fasciculus retroflexus) tracts of the habenular ganglion, also showed immunopositivity in these stages. Interestingly, the highest level of labeling was observed in premetamorphic larvae, just prior to entering the metamorphic stage. These data indicate that reelin expression is also prominent in brains of primitive vertebrates without layered cortical regions, suggesting that some physiological roles of reelin not related to the regulation of neuronal migration in layered cortical regions (i.e. involvement in axon pathfinding, synaptogenesis, dendritic arborization and neuronal plasticity) might have appeared earlier in evolution. PMID- 11861136 TI - Lectures. PMID- 11861137 TI - S1.Personality disorders and five personality factors in Europe and the USA. PMID- 11861138 TI - S2.Temperament in children and adolescents: evaluation and links with pyschopathology assessment. PMID- 11861139 TI - S4.The borderline concept from an international perspective. PMID- 11861141 TI - S6.Genetics and neurobiology of personality disorders. PMID- 11861140 TI - Sujets presentant une addiction et des troubles de la personnalite : aspects psychosociaux et traitement. PMID- 11861142 TI - S7.Interest and limits of the indirect methods for the assessment of personality disorders. PMID- 11861143 TI - S9.Group psychotherapy for personality disorders. PMID- 11861145 TI - S11.Cognitive and metaognitive strategies in personality disorders. PMID- 11861144 TI - S10.Emergency psychiatry and borderline disorders. PMID- 11861147 TI - S13.Assessment of personality disorders. PMID- 11861146 TI - S12.Personality disorder assessment and the MMPI-2: issues, scales and strategies. PMID- 11861148 TI - S14.Temperament and genetics. PMID- 11861149 TI - S15.Defense mechanisms and development. PMID- 11861150 TI - S16.Cluster C disorders: genetics, co-morbidity, course and outcome. PMID- 11861151 TI - S17.Personality inventories: an update. PMID- 11861153 TI - S19.Follow-up studies. PMID- 11861152 TI - Attachements et aspects fondamentaux. PMID- 11861154 TI - S20.Impulsivity, conduct disorders, eating disorders. PMID- 11861156 TI - S22.Paper sessions. PMID- 11861155 TI - S21.Pathological narcissism and psychopathy. PMID- 11861158 TI - S24.Personality and the elderly. PMID- 11861157 TI - S23.Personality disorders and violence. PMID- 11861160 TI - Patterning centers, regulatory genes and extrinsic mechanisms controlling arealization of the neocortex. AB - The adult mammalian neocortex, the major region of the cerebral cortex, is divided into functionally specialized areas, defined by distinct architecture and axonal connections. Extrinsic influences, such as thalamocortical input, and genetic regulation, intrinsic to the dorsal telencephalon, control the gradual emergence of area-specific properties during development. Major recent advances in this field include: the first demonstration of the genetic regulation of arealization, implicating the transcription factors Emx2 and Pax6 in the direct control of area identities; and the demonstration of the potential role of the signaling protein, fibroblast growth factor 8, in the early patterning of arealization genes, such as Emx2. PMID- 11861161 TI - Molecular mechanisms underlying cell fate specification in the developing telencephalon. AB - The cellular properties of neural progenitor cells have been best characterized in the telencephalon, the most complex region of the vertebrate brain. In recent years, several transcription factors, including Mash1, Ngn1/2, Pax6 and Emx1/2, and signaling molecules, such as Notch and bone morphogenetic proteins, have emerged as important players in key areas of telencephalic development. These include the specification of positional identity, the proliferation of neural stem cells and their commitment to a neuronal or glial fate, and the differentiation of layer-specific neuronal phenotypes in the cerebral cortex. PMID- 11861162 TI - Revisiting cell fate specification in the inner ear. AB - Generating the diversity of cell types in the inner ear may require an interplay between regional compartmentalization and local cellular interactions. Recent evidence has come from gene targeting, lineage analysis, fate mapping and gene expression studies. Notch signaling and neurogenic gene regulation are involved in patterning or specification of sensory organs, ganglion cells and hair cell mechanoreceptors. PMID- 11861163 TI - Patterning the neural retina. AB - The early patterning events that shape the neural retina guide the genesis and distribution of postmitotic cell types, as well as their connectivity. The recent discovery of key signaling pathways and transcription factors involved in establishing central, anterior-posterior, and dorsal-ventral retinal patterning has given us insights into the molecular mechanisms controlling these events. PMID- 11861164 TI - Neurodevelopmental control by thyroid hormone receptors. AB - Recent studies have provided insights into the neurodevelopmental functions of thyroid hormone signaling. The nuclear thyroid hormone receptors (TRs) are ligand activated transcription factors and a variety of TR isotypes, generated by two genes, mediate distinct processes. In addition, deiodinase enzymes that regulate levels of the main active form of thyroid hormone, T3, are likely to cooperate closely with TRs in specifying a localized and timely response to thyroid hormones in target tissues. Some of the most sensitive processes controlled by these pathways are in the auditory and visual sensory systems. PMID- 11861165 TI - Sonic hedgehog in the nervous system: functions, modifications and mechanisms. AB - Signaling by Sonic hedgehog (Shh) controls important developmental processes, including dorsoventral neural tube patterning, neural stem cell proliferation, and neuronal and glial cell survival. Shh signaling involves lipid modifications to Shh itself, as well as changes in protein subcellular localization. Recent advances have revealed the importance of palmitoylation and acylation of Shh on its potency and migration capacity. Subsequent trafficking and organelle sorting in the Shh signaling pathway have been observed; these observations offer a new dimension to our understanding of downstream signal transduction events. PMID- 11861166 TI - Variations on the Notch pathway in neural development. AB - Notch signaling allows cells in contact to adopt different fates. Regulation of the Notch pathway allows for the same signaling mechanism to be used in a wide variety of contexts during development. Intracellular activities of the E3 ubiquitin ligases Sel-10 and Neuralized involve proteasome-dependent degradation in the regulation of Notch pathway activity. Extracellular manipulations of Notch by Fringe and Scabrous regulate the pathway by changing Notch interactions outside the cell. These regulatory mechanisms, along with many others, affect how Notch signaling activity influences cell fate determination. PMID- 11861167 TI - Synaptogenesis: insights from worm and fly. AB - Synapse formation is the ultimate step in wiring a nervous system. Synapses are remarkably diverse in size and shape, and are regulated dynamically. Recently, live observations combined with ultrastructural analysis have revealed many details of the cellular interactions that precede synapse formation. Genetic screens in Caenorhabditis elegans and Drosophila have implicated signaling pathways that may involve small G-proteins, ubiquitin-mediated protein degradation and selective cell adhesion in target recognition, synaptic assembly and growth. PMID- 11861168 TI - Development of neuronal connectivity in Drosophila antennal lobes and mushroom bodies. AB - Recent advances in the study of the connectivity of Drosophila olfactory system include the demonstration that olfactory receptor neurons project to specific glomeruli according to the receptor type they express, and that their projection neuron partners are prespecified to innervate particular glomeruli by birth order or time. This same theme of sequential generation has been observed in the generation of the three major types of mushroom body neurons. PMID- 11861169 TI - Wiring the zebrafish: axon guidance and synaptogenesis. AB - Many zebrafish mutants have specific defects in axon guidance or synaptogenesis, particularly in the retinotectal and motor systems. Several mutants have now been characterized in detail and/or cloned. A combination of genetic studies, in vivo imaging and new techniques for misexpressing genes or blocking their function promises to reveal the molecules and principles that govern wiring of the vertebrate nervous system. PMID- 11861170 TI - Activity-dependent mapping in the retinotectal projection. AB - It is now 15 years since the discovery that N-methyl-d-aspartate receptor activity is required to maintain the refined topographic organization of retinotectal projections. Recent studies have identified additional components of the signaling pathways required for activity-dependent map formation and maintenance. Nitric oxide and brain-derived neurotrophic factor, candidate retrograde messengers, and serotonin and acetylcholine, modulators of neuronal excitability, all affect mapping. These studies indicate that the mapping process intersects with other processes fundamental to visual system development and function, such as process outgrowth, synaptic turnover and neuromodulation. PMID- 11861171 TI - Patterning of skeletal muscle. AB - Recent studies challenge the view that signals provided by motor neurons are required to activate subsynaptic nuclei and induce postsynaptic specializations in developing skeletal muscle. New findings show that acetylcholine receptor genes are expressed and that acetylcholine receptor clusters form preferentially in the prospective synaptic region of muscle independently of motor innervation. These results indicate that developing myotubes are patterned by mechanisms intrinsic to developing muscles and raise the possibility that patterning of muscles may influence the growth pattern of motor axons and the sites where synapses form. PMID- 11861172 TI - Ocular dominance development revisited. AB - New approaches to the study of ocular dominance development, a model system for the development of neural architecture, indicate that eye-specific columns in primary visual cortex emerge substantially before the onset of the critical period, during which neural connections can be altered by visual experience. The timing, speed and specificity of column emergence implicate molecular patterning mechanisms, along with patterns of neural activity, in the generation of this columnar architecture. PMID- 11861173 TI - Applying genomics technologies to neural development. AB - Genomic technologies have evolved from a minority interest to a set of generally applicable, powerful tools. Recent studies have demonstrated that such tools are of great use in studies of neural development, particularly when allied to advances in data analysis and methods for analyzing gene function. PMID- 11861176 TI - Matters temporal. AB - Current evidence suggests that neural Hebbian learning in cortical and hippocampal synapses is fundamentally predictive rather than conventionally correlational. Much attention is focussing on what sort of predictions are acquired, and in what neural architectures. A recent paper by Rao and Sejnowski has suggested an interesting interpretation in terms of a popular predictive algorithm that has roots in psychology, computer science and engineering. PMID- 11861174 TI - Maternal infection: window on neuroimmune interactions in fetal brain development and mental illness. AB - Direct viral infection of the developing brain can have disastrous consequences for the fetus. More subtle and perhaps more insidious are viral infections of the pregnant mother, which can have long-lasting effects such as an increased risk of schizophrenia in the offspring. A recent mouse model has shown that respiratory infection in the pregnant mother leads to marked behavioral and pharmacological abnormalities in the offspring, some of which are relevant for schizophrenia and autism. This effect on fetal brain development might be caused by the maternal antiviral immune response, possibly mediated by cytokines. PMID- 11861177 TI - Neuroecology and psychological modularity. PMID- 11861178 TI - A reply to the defenders of the faith. PMID- 11861179 TI - Breakfast in the Pacific. AB - The Sixth Natural Language Processing Pacific Rim Symposium was held on 27--30 November 2001, at the National Center of Science, Tokyo, Japan. PMID- 11861180 TI - Inching ahead, artificially. AB - The inaugural Australian workshop in Artificial Life took place on the 11th of December 2001 at Adelaide University in South Australia. PMID- 11861181 TI - Finding maps in the cortex. PMID- 11861182 TI - Brain activity around the clock. PMID- 11861183 TI - Finding order in schizophrenic thought disorder. PMID- 11861185 TI - One day, as I was lying in the scanner... PMID- 11861184 TI - Knowing our imitations. PMID- 11861187 TI - What competition? PMID- 11861188 TI - What competition? PMID- 11861189 TI - Visual masking and RSVP reveal neural competition. AB - A test visual stimulus is harder to recognize when another stimulus is presented in close temporal vicinity; presenting stimuli in close spatial vicinity of a test stimulus reduces its visibility; presenting a stimulus to one eye can render invisible another stimulus presented to the other eye; and perceiving one interpretation of an ambiguous image prevents the simultaneous perception of other visual interpretations. A single, neurophysiological theory, which may be called 'neural competition' might explain all these phenomena: when two alternative neural visual representations co-exist in the brain, they compete against each other. PMID- 11861190 TI - Constraining the neural representation of the visual world. AB - Understanding the perception of all but the most impoverished and artificial scenes presents a different and probably far greater challenge from understanding face recognition, reading, or identification (or even categorization) of single objects. Central issues in the interpretation of structured objects and scenes are reviewed, starting with fundamentals such as the meaning of seeing. A theoretical approach to this formidable task is outlined, motivated by some recent developments in neuroscience and neurophilosophy. PMID- 11861191 TI - Categories, prototypes and memory systems in Alzheimer's disease. AB - Recent studies indicate that category learning is mediated by multiple neuronal systems. It has been shown that patients with marked impairments in executive functions, explicit memory and procedural learning can categorize the exemplars and prototype of a previously trained category. Simple, self-organizing neuronal networks can explain prototype learning and related dysfunctions in Alzheimer's disease, and provide a model of how prototype learning is mediated by circumscribed mechanisms in the visual cortex. PMID- 11861192 TI - Perceptual adaptation: motion parallels orientation. AB - Adaptation phenomena provide striking examples of perceptual plasticity and offer valuable insight into the mechanisms of visual coding. Within the context of recent progress in neurobiology and computational modelling, I review evidence from studies employing psychophysical adaptation to investigate orientation and motion processing. These studies reveal marked similarities between the orientation and motion domains, raising the possibility that common computational principles underlie the processing of orientation and motion despite apparently distinct cortical substrates. PMID- 11861193 TI - Musical Jabberwocky? PMID- 11861194 TI - Tools for recognizing faces. PMID- 11861197 TI - [Prognostic factors of pregnancy and delivery complications in Senegalese adolescents and their newborn]. AB - Developing countries exhibit the highest adolescent fertility rate in the world. Undesirable outcome frequency during adolescents' pregnancy and delivery is debated. The aim of this study is to determine the incidence of these events and the factors predicting their occurrence among pregnant Senegalese adolescents and their newborn. The study was conducted between August 1st, 1999 and July 31st, 2000 in a national sample of 435 women attending prenatal clinics after a 20-week gestation. The data were collected by interviews: sociodemographic characteristics, obstetrical antecedents, current pregnancy history before inclusion in the study; the clinical characteristics were determined at inclusion. Then pregnancy and delivery complication occurrences were observed for mothers and their newborn during follow-up. Logistic regression analyses were performed to determine the factors associated with each complication, controlling the effects of other sociodemographic and clinical factors. The most frequent pregnancy complications were pregnancy-induced hypertension and toxaemia (17.5%). Anemia occurred in 25% of the women. At delivery, at least one of dystocia, eclampsia, or placenta haemorrhage complications occurred in 46% of women. The caesarian rate was 20% and the mortality rate was 71%. When planned, caesarian interventions lowered mortality. Forty percent of the newborns presented a neonatal distress while 17.9% of them weighed less than 2,500 g. The pregnancy induced hypertension and toxaemia incidence rates were higher when the weight at onset of pregnancy was unknown (OR = 3.7 [1.9-7.5] for hypertension and 3.1 [1.3 7.6] for toxaemia). When a woman with a narrow pelvis was less than 1.50 m tall, the delivery was more frequently assisted by forceps and extracting action in breech delivery. Dystocia, eclampsia, placenta haemorrhage and premature delivery were associated with high mortality. Overall, complications were not more frequent in the youngest (16 years and below), as suggested in previous studies. Those results suggest that undesirable events occur frequently during adolescents' pregnancy and delivery, and in their newborn, while attendance in prenatal clinics is adequate. The prognostic factors of these complications are identified. These complications can be avoided by improving ante- and perinatal care quality. PMID- 11861198 TI - [Obstetric catastrophes barely just avoided: near misses in Moroccan hospitals]. AB - A consensus definition of obstetric catastrophes barely only just avoided, called near miss cases in the recent scientific literature, has been elaborated during an international seminar held in Morocco. A near miss case was defined as "any pregnant or recently delivered - or aborted - woman, whose immediate survival is threatened and who survives by chance or because of the hospital care received". This definition was then operationalised using severity criteria combining clinical signs and types of intervention clear enough to easily screen near miss cases in hospital files. These criteria were then used to identify the near misses that occurred in 1998 in two public Moroccan hospitals (Tetouan and Sidi Kacem). A total of 81 cases of severe maternal complications (76 near misses and 5 deaths) were collected, a frequency of 11.9% among hospital admissions for delivery or pregnancy complications. The interest and limitations of such a near miss case definition are discussed. It seems that the criteria applied were operational in the Moroccan context. They are specific, i.e. they permitted to identify true cases of mother's life threatening complications. Finally, they generated a sufficiently great number of cases and a range of situations large enough to hold monthly audits and to identify sub-standard care. PMID- 11861195 TI - [Investigation of a shigellosis outbreak in a rural zone of Senegal]. AB - The investigation of this outbreak took place in Niakhar (IRD project study area), a rural area located 150 km East of Dakar. Covering 30 villages, with a population of about 30,000 inhabitants, the area has been under demographic and epidemiological surveillance. In 1999, the surveillance allowed 973 cases of shigellosis to be recorded out of 1,751 cases of dysenteric diarrhoeas. The outbreak reached a peak during the raining season (73% of the cases in September and October). All the villages and all age groups were concerned. The attack rate in the population was 3%. During the same period, 22 deaths were attributed to shigellosis, leading to a lethality of 2.3%. Children under five were the most affected. The bacteriological examination isolated S. flexneri in 72% of 12 stools samples and S. dysenteriae A1 (SD1) in 14%. All serotypes were resistant to ampicillin and susceptible to quinolones. Susceptibility to cotrimoxazole was unconstant. Apart from the usual factors involved in the spread of diarrhoeas, the main reason evoked to explain the duration of the epidemic has been the lack of adequate emergency antibiotherapy treatment. Quinolones, recommended by the health authorities, could not be provided by health services. The identification of the SD1 serotype confirmed its involvement in the outbreak of shigellosis. The exact role of S. flexneri in shigellosis epidemics should be further studied. PMID- 11861199 TI - [Hereditary factors in strabismus]. AB - This retrospective study took place in the Ophthalmology Department of Douala General Hospital. The aim was to determine the importance of hereditary factors in a group of strabismic Cameroonians. In a series of 275 patients with strabismus, we found 79 familial cases (28.72%). Among them, 22.78% had more than one relative with squint and 75% had the same type of strabismus as their relative. There was no significant difference in the percentage of familial cases with regard to the type of strabismus, the sex, the mode of fixation and the impairment of ductions. As far as ametropias are concerned, only myopia showed a hereditary tendency in our series. The authors agree with the literature in that there is a significant hereditary component in the cause of strabismus, but its genetic sites are yet to be identified. There is strong hope in this direction with the decoding of the human genom and the advances in molecular biology. However, the study of familial cases is important since it allows high risk groups to be defined and screened. It thus makes it possible to successfully fight amblyopia through early detection and treatment. PMID- 11861200 TI - [Maternal risk factors and low birth weight in Senegalese teenagers: the example of a hospital centre in Dakar]. AB - AIMS OF THE STUDY: a) to identify the risk factors associated with newborn babies' low birth weight in teenage mothers; b) to propose prevention strategies for lower-ing neonatal morbidity and mortality. SPHERE OF THE STUDY: the study was carried out at the maternity and neonatology service of the Abass Ndao hospital centre, a urban community hospital located in the Southern part of Dakar. MATIERIAL AND METHODS: this retrospective study was carried out between July 1, 1998 and June 31, 1999. All new mothers under 20 who had given birth to living newborn babies have been included in the study and categorised into two groups, according to their babies' birth weight: 1) a first group of teenagers whose newborn babies' birth weight was lower than 2,500 g (low birth weight); 2) a second group constituted of women whose newborn babies' birth weight was higher than 2,500 g, and which was used as a control. The socio-demographic, biometrics, maternal and obstetric factors have been analysed and compared. RESULTS: out of the 4,586 women in childbirth during the study period, 456 were under 20 years of age, which corresponds to a prevalence rate of 10%. One hundred and five women had newborns weighing less than 2,500 g, which corresponds to a prevalence rate of 23%. Certain factors were found to be significantly associated with low birth weight: low weight gain during pregnancy (p = 0.04), fewer antenatal consultations (0.006), and kidney-related syndromes during pregnancy (0.0005). CONCLUSION: The results of that study allow us to recommend the following strategies: - control and improvement of nutritional behaviour during pregnancy; campaigning for a better attendance at antenatal consultation services for the early detection of pathologies during pregnancy and for preventing kidney-related syndromes. PMID- 11861201 TI - [Hypothyroidism: clinical and etiological aspects in Cotonou (Republic of Benin)]. AB - To evaluate the frequency of clinical manifestations and to study the etiological aspects of hypothyroidism in hypothyroid patients in Cotonou, we carried out a retrospective analysis of the medical reports on hypothyroid patients followed in our clinical practice. A total of 33 patients was thus studied, comprising 8 men (24%) and 25 women (76%) with mean age of 45,8 years for men and 40,4 years for women. The more frequent clinical manifestations observed were: face edema (45%), weight gain (45%), paresthesia (42%), fatigue (39%), lethargy (30%) and bradycardia (24%). Constipation (12%), sensation of cold (9%), depilation (6%) and dry skin (6%) were less frequently observed. Myalgia, hoarseness and menstrual irregularities were present in 15% of the cases respectively. Regarding the etiology, 82% of the cases were primary hypothyroidism and only 18% were of central origin. Thyroidectomy was the leading cause in our hypothyroid patients, representing 70% of all cases and 85% of primary hypothyroidism. Radioiodine treatment and autoimmune thyroiditis were equally found in 6% of the cases. Central hypothyroidism was related to a pituitary adenoma in four cases (12%) and to Sheehan syndrome in two cases (6%). As it can be expected, hypercholesterolemia was present in 82% of the patients but creatine phosphokinase elevation was more frequent (94% of the patients). Compared to the data reported in the literature, the frequency of the symptoms and signs of hypothyroidism seems to be underevaluated in our study and the frequency of autoimmune thyroiditis as a cause of hypothyroidism is low. PMID- 11861202 TI - [Detection of meningococcal meningitis epidemics in Africa: a new recommendation]. AB - In sub-Saharan Africa, the control of meningococcal meningitis epidemics relies on early epidemic detection and mass vaccination. However, experience shows that interventions are often initiated too late to have a significant impact on the epidemic. A new recommendation drafted by participants of a consensus meeting proposes an alert threshold and an epidemic threshold based on the weekly number or incidence of meningitis cases, according to the population size and the epidemic risk, resulting in indicators with high sensitivity and specificity for the detection of an emerging epidemic. Meningitis outbreak investigations must include an assessment of the quality of epidemiologic surveillance. The new recommendation is published in English and French in the Weekly Epidemiologic Record [12]. The success of this consensus meeting shows the value of integrating results from surveillance, field experience and operational research for designing new health strategies. PMID- 11861204 TI - [The delivery record: a help to make the right decision]. AB - The delivery record is a key document for risk assessment and proper decision making at the right time during delivery. If it is exhaustive or badly presented, health workers see it as an administrative constraint, not as a help. The authors recall some principles regarding its form and contents. Its different parts are reviewed: administrative data, medical history, antenatal care, admission clinical examination, partograph, delivery, newborn, recent post-partum and exit examinations. An example of a delivery record used in Nouakchott is presented. PMID- 11861203 TI - [Treatment of visceral leishmaniasis: efficacy and limits of miltefosine]. PMID- 11861205 TI - [AIDS and social situations of women in Africa: from notions to research methods]. AB - When researchers or actors intend to analyse women's situations in the context of AIDS in Africa, they are conducted to take into account the whole context these women live in: their access to care and prevention, their ability to make choices in their emotional, sexual and marital life, their access to education and to a gainful employment. It is also important in such an approach to define precisely the notions used. This paper proposes first to describe the several realities existing under the notions of vulnerability, autonomy, negotiation, sexuality, prostitution, motherhood and mother and child health. Through examples from researches conducted in Cote d'Ivoire and in other African countries, we enlighten how stereotypes are still used to describe women's situations in Africa. We insist on the need to propose fine-tuned and balanced analyses, in order to avoid sweeping generalisations that are harmful to women themselves. Secondly, we describe the methodological tools that can be used in this kind of analysis of notions and contexts in gender relations. We describe qualitative and quantitative survey techniques. For each one, we emphasise their requirements but also their effects on our research process on women's experiences facing AIDS. PMID- 11861206 TI - [Autologous blood injections for treating hypotonies after trabeculectomy]. AB - The authors carried out a retrospective study in order to assess the efficacy of intrabled autologous blood injections after trabeculectomy. The indication for treatment was hypotony associated with overfiltration. Twelve eyes of 12 patients including seven men (58.3%) and five women (41.67 %) underwent from one to four (mean 1.7) subconjunctival injections. The age of the patients ranged from 31 to 66 years (mean 52.4 years). All the patients were diagnosed with open-angle glaucoma. Three eyes underwent trabeculectomy with mitomycine C, one with 5 fluorouracil and eight with no antimetabolite. The mean post-needling period was 12.3 months (ranging from 7 to 28 months). After intrabled blood injections, the average intraocular pressure increased from 2.7 1.2 mmHg (ranging from 0 to 6 mmHg) to 8.2 4.2 mmHg (ranging from 4 to 16 mmHg). The difference was statistically significant (P < 0.5). After treatment, the average visual acuity increased from 1.8/10 to 3.2/10. This difference was not statistically significant (P > 0.5). However, the procedure was ineffective in two patients (16.7%) as regards intraocular pressure and in seven patients (58.3%) as regards visual acuity. Hyphema, the most frequent complication (58.3% of our cases) is usually small, transient, and without sequelae. Although it may be delayed, it may be important and it induce intraocular hypertony (10% of our cases) or it may be associated with intravital blood. PMID- 11861208 TI - Bi-directional coupling between dihydropyridine receptors and ryanodine receptors. AB - The control of calcium signaling between plasma membrane dihydropyridine receptors (DHPRs or L-type calcium channels) and ryanodine receptors (RyRs or calcium release channels) located in the endoplasmic/sarcoplasmic reticulum (ER/SR) underlies a broad array of functions including skeletal muscle contraction, cardiac performance, arteriole tone, neurosecretion, synaptic plasticity, and gene regulation. It has long been appreciated that DHPR activation of RyRs and subsequent calcium release from intracellular stores represents a key event in the control of these processes. In excitable cells, DHPRs trigger the release of intracellular calcium by promoting the opening of nearby RyRs (termed orthograde coupling). Interestingly, the signaling interaction between DHPRs and RyRs is often bi-directional such that the calcium conducting activity of DHPR channels is also regulated by its interaction with the RyR (termed retrograde coupling). Recent data indicate that skeletal, cardiac, and neuronal cells utilize fundamentally distinct DHPR/RyR bi directional coupling mechanisms (chemical and mechanical) in order to control the efficiency, fidelity, and activity of each of these two essential calcium channels. This review will focus on evaluating the nature and molecular determinants of these coupling mechanisms, as well as the extent to which excitable cell function is influenced by bi-directional DHPR/RyR calcium signaling. PMID- 11861209 TI - Mechanisms of autoinhibition in cyclic nucleotide-dependent protein kinases. AB - Cyclic AMP-dependent protein kinase (PKA) and cGMP-dependent protein kinase (PKG) are autoinhibited through multiple interactions between their respective regulatory and catalytic domains. A large portion of this autoinhibition occurs through interactions between residues within the catalytic domain and those within either a substrate-like sequence (-RRXSX-) or pseudosubstrate sequence ( RRXAX-) in the regulatory domains. These contacts effectively inhibit catalysis by blocking substrate binding. Particularly important contacts involve the P-2, P 3, and P+1 residues where either serine, which is potentially autophosphorylated, or alanine occupies the P0 position. The primary sequence is apparently less important for autoinhibition in PKGs than in PKAs, and a conserved serine at P+2 in PKGs is important for autoinhibitory contacts. Elements outside the substrate related sequences also contribute to autoinhibition in both PKA and PKG. For example, synthetic peptides with relatively short pseudosubstrate sequences are weak inhibitors; while heat-denatured RII subunit does not inhibit catalytic subunit, it is still rapidly autophosphorylated; and truncated PKGs lacking the substrate-like sequence are still partially autoinhibited. Thus, capacity for autoinhibition of PKA or PKG is provided by contacts involving direct interactions with the catalytic site and by contacts that stabilize an inactive conformation. PMID- 11861210 TI - Spiroplasmas: evolution, adaptation and diversity. AB - Since its designation as a separate genus some 30 years ago, Spiroplasmas have been well documented in a wide range of hosts and as the causative agent of several plant and insect diseases. One major area of research is the continued identification and taxonomical characterization of new Spiroplasma sp. combined with a determination of phylogenetic relationships among the various Spiroplasma sp. and between the Spiroplasmas and other members of the Mollicutes and Eubacteria. Although most phylogenetic analyses have been dependent on 16S rDNA sequence data, progress in two Spiroplasma sp. genome sequencing projects will provide new genomic regions for comparative focus. The co-evolution of Spiroplasmas with their arthropod hosts has provided an additional research focus to study host specificity and attachment. The diversity of symbiotic relationships between Spiroplasmas and their hosts has led to the study of commensal, mutualistic, and pathogenic relationships. Pathogenesis in insect hosts or in plants, transferred by insect hosts, is a major research focus, which requires attachment and invasion into insect tissues beyond the initial infection site, and successful movement to other tissues. The diversity and adaptations that have occurred during the evolution of the Spiroplasmas with their hosts will be the primary focus of this article. PMID- 11861211 TI - Chlamydia pneumoniae as a respiratory pathogen. AB - Chlamydia pneumoniae is a recently recognized human respiratory pathogen with a unique biphasic life cycle characterized by an obligate intracellular (replicative) and an extracellular (infectious) form of the organism. C. pneumoniae is widely distributed and, via the respiratory route, infects the majority of the world's population. The majority (70%) of acute human C. pneumoniae respiratory tract infections are asymptomatic or only mildly symptomatic but a minority (30%) cause more severe respiratory illnesses including community-acquired pneumonia, bronchitis and a variety of upper airway illnesses. After acute infection the C. pneumoniae intracellular life cycle is characterized by the development of metabolically inert (and thus antibiotic resistant) atypical "persistent" inclusions; this biologic behavior correlates with a clinical course following acute symptomatic illness that is characterized by persistence of symptoms that are difficult to treat with antibiotics. A role for C. pneumoniae in chronic respiratory illness is currently under investigation: "persistent" intracellular inclusions contain increased quantities of chlamydial heat shock protein 60 (hsp 60), a highly immunogenic protein that has been implicated in the pathogenesis of established chronic inflammatory chlamydial diseases (blinding trachoma, pelvic inflammatory disease and tubal infertility). An emerging body of evidence, including host immune response to chlamydial hsp 60, links C. pneumoniae infection with a spectrum of chronic inflammatory lung diseases of currently unknown etiology (asthma, chronic bronchitis and chronic obstructive pulmonary disease (COPD)). Further laboratory developments, including reliable and practical diagnostic methods and antibiotics effective against persistent infection, will be required to recognize and treat acute C. pneumoniae infection, and to advance our knowledge and understanding of the role of chronic infection in asthma, chronic bronchitis and COPD. PMID- 11861212 TI - Regulation of calcium release by interdomain interaction within ryanodine receptors. AB - In excitation-contraction (E-C) coupling, various types of activation signals, which are received presumably at the bulky cytoplasmic domain of the ryanodine receptor (RyR), are translated (or transduced) into the opening of the Ca2+ release channel located in the trans-membrane domain of the RyR. In order to elucidate the detailed mechanism of the signal transduction process, it is essential (i) to identify various sub-domains of the RyR that are involved in the Ca2+ channel regulation, (ii) to characterize the events occurring in these sub domains during the activation process, and (iii) to characterize the modes of active interactions among these sub-domains. Recent developments in the E-C coupling research have provided us with new insight into each of these aspects, as outlined in this review. Of many putative regulatory sub-domains of the RyR, two domains (designated as N-terminal domain and central domain) are particularly interesting, because disease-linked mutations that have occurred in these domains (malignant hyperthermia and central core disease in skeletal muscle, and inheritable cardiac disease) induce abnormal modes of Ca2+ channel regulation. Pieces of evidence accumulated to this date suggest the following hypothesis. The N-terminal and central domains form, at least partly, the interacting domain pair, and unzipping and zipping actions of such domain-pair are involved in the opening and closing actions of the Ca2+ channel, respectively. We also propose that there are local conformational changes in the signal reception domains (e.g. the II-III loop-binding core), and such conformational changes are coupled with the aforementioned actions of the interacting domain pair. It seems that by virtue of such a coordination of the events occurring in various regions of the RyR, the Ca2+ channel can recognize the activation signal received at the cytoplasmic region of the RyR. PMID- 11861213 TI - The KSHV latency-associated nuclear antigen: a multifunctional protein. AB - Kaposi's sarcoma(KS)-associated herpes virus (KSHV) or human herpesvirus 8 (HHV 8) is highly associated with KS, primary effusion lymphoma (PEL), and multicentric Castleman's disease, an aggressive lymphoproliferative disorder (1 3). Most tumor cells are latently infected with KSHV in which a small subset of viral genes are expressed (4-6). Of these latently expressed genes, the latency associated nuclear antigen (LANA1, LNA, or LNA1) is the only protein consistently shown to be highly expressed by in situ hybridization and immunohistochemistry (7 10). In the past few years multiple functions have been demonstrated for LANA1. Here we review LANA1's roles in KSHV infection. Topics discussed include LANA1's roles in episome persistence, regulation of transcription and interaction with cellular proteins. PMID- 11861214 TI - An integrated view of the roles and mechanisms of heat shock protein gp96-peptide complex in eliciting immune response. AB - Heat shock protein (HSP) gp96, or grp94 is an endoplasmic reticular (ER) paralog of the cytosolic HSP90. Being abundant and non-polymorphic, gp96 plays significant roles in maintaining protein homeostasis in the secretory pathway. This "house-keeping" role of gp96 has now been overshadowed by the intriguing findings that gp96 modulates both the innate and adaptive components of the immune system. It has been found that, (i) gp96 is one of the major peptide binding proteins in the ER, (ii) gp96 interacts specifically with receptors including CD91 and possibly toll-like receptors (TLRs), on the surface of professional antigen presenting cells (APCs), (iii) interaction with APCs leads to re-presentation of gp96-chaperoned peptides to the major histocompatibility complex (MHC) molecules of APCs, (iv) direct access of gp96 to APCs triggers functional activation of APCs. In this review, we will examine each of these immunological attributes of gp96 critically. As experimentalists, we will also propose specific experiments to examine the argument that gp96, perhaps along with other members of HSP family, is the antigenic carrier for mediating cross priming of antigen-specific T lymphocytes in vertebrates. PMID- 11861215 TI - The role of human papillomaviruses in human cancers. AB - Of the more than 100 different human papillomavirus types (HPVs), the "high-risk" HPVs are associated with the vast majority of cervical carcinoma, and a pathologically distinct group of oropharyngeal tumors. In addition, other HPVs are associated with cutaneous tumors, in particular epidermodysplasia verruciformis and non-melanoma skin cancers. In general, HPV-associated cancers arise from a single accidental integration event of the viral genome into a host cell chromosome. Integration is a terminal event for the viral life cycle. Even though integration does not occur at specific chromosomal hot spots in the human genome, it follows a consistent pattern with respect to the viral genome, and expression of the HPV E6 and E7 genes is consistently retained. The normal function of E6 and E7 is to establish and maintain a cellular milieu that allows for viral genome replication. E6 and E7 target important cellular growth regulatory circuits among them the p53 and retinoblastoma tumor suppressors, respectively. Uncontrolled expression of the E6 and E7 proteins, as a consequence of viral integration is paramount to the establishment and maintenance of the tumorigenic state. In addition, expression of E6 and E7 increases genomic instability of the host cell thus accelerating malignant progression. Taken together, there is compelling molecular and epidemiological evidence in support of an oncogenic function of certain HPVs. PMID- 11861216 TI - Borna disease virus infection of the neonatal rat: developmental brain injury model of autism spectrum disorders. AB - Autism spectrum disorders (ASD) have been the focus of a great deal of research and clinical speculation. This intense interest relates to both the perplexing pathogenesis and devastating consequences of these disorders. One of the obstacles to understanding the pathogenesis of autism and its efficient treatment has been the paucity of animal models that could be used for hypotheses-driven mechanistic studies of abnormal brain and behavior development and for the pre clinical testing novel pharmacological treatments. The present review provides a detailed analysis of a new animal model of ASD. This model utilizes neonatal Borna disease virus (BDV) infection of the rat brain as a unique experimental teratogen to study the pathogenesis of neurodevelopmental damage. For more than a decade, studies of the BDV animal model have yielded much insight into the pathogenic processes of abnormal brain development and resulting autistic-like behavioral abnormalities in rats. The most recent experiments demonstrate the utility of the BDV model for studying the pathophysiological mechanisms of the gene-environment interaction that determines differential disease outcomes and variability in responses to treatments. PMID- 11861217 TI - Structural interaction between RYRs and DHPRs in calcium release units of cardiac and skeletal muscle cells. AB - Excitation-contraction (e-c) coupling in muscle cells is a mechanism that allows transduction of exterior-membrane depolarization in Ca2+ release from the Sarcoplasmic Reticulum (SR). The communication between external and internal membranes is possible thanks to the interaction between Dihydropyridine Receptors (DHPRs), voltage-gated Ca2+ channels located in exterior membranes, and Ryanodine Receptors (RyRs), the Ca2+ release channels of the SR. In both skeletal and cardiac muscle cells the key structural element that allows DHPRs and RyRs to interact with each other is their vicinity. However, the signal that the two molecules use to communicate is not the same in the two muscle types. In the heart, the inward flux of Ca2+ through DHPRs, that follows depolarization, triggers the opening of RyRs (calcium induced calcium release). In skeletal muscle, on the other hand, Ca2+ is not needed for RyRs activation; instead the coupling between the two molecules involves a direct link between them (mechanical coupling). Ultrastructural studies show that functional differences can be explained by differences in the DHPR/RyR reciprocal association: whereas the two proteins are very close to each other in both muscles, DHPRs form tetrads only in skeletal fibers. Tetrads represent the structural DHPR/RyR link that allows Ca2+ independent coupling in skeletal muscle. PMID- 11861218 TI - Clinical implications of human papillomavirus infection. AB - Human papillomaviruses (HPV) are small DNA viruses associated with specific mucosal and epithelial lesions ranging from benign proliferative lesions to invasive carcinomas. Over 100 types of HPV have been identified, some of which are associated with benign lesions (low risk types) and others are associated with malignancies (high risk types). While the genome consists of 6 early genes and 2 late genes, the E6 and E7 genes have been most studied because they interact with p53 and Rb, respectively, thus contributing to the ability of HPV to mediate oncogenesis. Cervical carcinoma is the most common and most studied HPV-related malignancy. These lesions are thought to be originated from persistent high-risk type HPV infections which progress to well characterized precursor lesions and finally to carcinoma. This same HPV related progression has also been observed in other anogenital malignancies including anal, penile and vulvar carcinomas. Although the evidence is not as conclusive, HPV also likely plays a role in the development of a subset of squamous cell carcinomas of the head and neck as well as other cutaneous malignancies. While HPV infection is common, the progression to malignancy is relatively rare indicating a potential role for immune protection against persistent infection. This is supported by the fact that HPV infection and related malignancies are common in the immunosuppressed population. Thus, efforts have been placed on development of HPV vaccines to prevent and treat these common and diverse groups of HPV related malignancies. PMID- 11861219 TI - The Epstein Barr nuclear antigen EBNA3C regulates transcription, cell transformation and cell migration. AB - The Epstein-Barr virus (EBV) infects most of the human population and persists in B-lymphocytes for the lifetime of the host. During the establishment of latent infection a unique repertoire of genes are expressed. The EBV nuclear antigen EBNA3C is essential for growth transformation of primary B-lymphocytes in vitro. EBNA3C regulates the transcription of a number of viral and cellular genes important for the immortalization process. Interaction of EBNA3C with the cellular transcription factor RBP-Jkappa and HDAC1 modulates transcriptional activation. Additionally, EBNA3C disrupts the cyclin/cdk-Rb-E2F pathway that regulates cell cycle progression through the restriction point at G1. Recent studies showed that the carboxy terminal region of EBNA3C from aa 366-992, essential for the immortalization of primary B-cells, interacts with Prothymosin(alpha) (ProTalpha) and Nm23-H1. The interaction of EBNA3C with ProTalpha as well as the histone acetylase p300 suggested a possible role in modulation of histone acetylation and chromatin remodeling. Cell migration assays geared towards determining the effect of EBNA3C on Nm23-H1 antimetastatic function suggests that EBNA3C suppresses the effects of NM23-H1 on the motility of breast carcinoma as well as Burkitt's lymphoma cells. This observation suggests that EBNA3C may be involved in driving the metastatic process in EBV associated human malignancies. PMID- 11861220 TI - Herpes simplex virus. AB - Herpes simplex virus (HSV) commonly causes human infections in the orofacial region (HSV-1) and in the genital region (HSV-2). Productive viral infection in mucosal epithelial cells may result in clinical symptoms and is followed by a latent infection within sensory neurons. During productive infection a large number of viral gene products are expressed while during latent infection few or no viral proteins are expressed. Reactivation from latency results in recurrent infections and disease at or near the primary site of infection. Understanding the details of the two stages of the HSV life cycle is a particular focus of current research on HSV. The virus interacts with and modifies numerous host cell functions in both epithelial and neuronal cells, and studies of HSV have enhanced our knowledge of many fundamental processes in eukaryotic cells. Ongoing research continues to uncover novel effects of HSV on cells, and a complete understanding of HSV infection during both productive and latent infection should allow the design of new antiviral agents and vaccines and increased knowledge of basic cell and molecular biology. This review article is designed to provide an introduction to HSV biology and key aspects of the infection cycle. PMID- 11861221 TI - Molecular aspects of scleroderma. AB - Scleroderma is a complex disease characterized by activation of the immune system, small-vessel vasculopathy, and fibrosis of the skin and other organs. This review is limited to the discussion of scleroderma fibroblast biology and the cellular and molecular mechanisms that contribute to the abnormal deposition of collagen. Selected aspects of abnormal extracellular matrix (ECM) regulation by scleroderma fibroblasts are critically reviewed. These include the role of transforming growth factor-beta (TGF-beta) and connective tissue growth factor (CTGF) and their receptors in the fibrotic process in scleroderma and the overview of the transcription factors involved in regulation of the human alpha2 (I) collagen (COL1A2) gene. PMID- 11861222 TI - Photoimmune suppression and photocarcinogenesis. AB - The primary cause of non-melanoma skin cancer, the most prevalent form of human neoplasia, is the ultraviolet (UV) radiation found in sunlight. Exposing mice to UV radiation induces skin cancers that are highly antigenic. Upon transfer of an UV-induced skin cancer to a normal syngeneic mouse, the tumor cells are recognized and rapidly destroyed by the immune system of the recipient. This raises the question of how these cancers avoided immune destruction during their development in the UV-irradiated host. This question was answered when it was discovered that in addition to being carcinogenic, UV radiation was also immunosuppressive. Studies with immune suppressed transplantation recipients, and biopsy proven skin cancer patients have confirmed that UV-induced immune suppression is a risk factor for skin cancer development in humans. It is of great importance, therefore, to understand the mechanisms underlying UV-induced immune suppression. The focus of this manuscript will be to use some examples from the more recent scientific literature to review the mechanisms by which UV radiation suppresses the immune response and allows for the progressive outgrowth of antigenic skin tumors. PMID- 11861223 TI - The historical aspects of photocarcinogenesis. AB - The concept of photocarcinogenesis is of fairly recent duration. Although cancer of the breast is described in the ancient Greek medical literature, skin cancer is not mentioned even as late as the 18th Century. This is most likely due to the poor survival of humans, 80% of people did not live past 40 years, and only 6% lived longer than 60 years. The first association of skin cancer (face and lip) with outdoor exposure dates to the middle of the 19th Century. About that time it was shown that it was Ultraviolet Radiation (UVR) that could cause skin and eye inflammation. It was not until the 20th Century that competent epidemiologic studies associated human skin cancer with chronic solar exposure, and it was shown that chemicals could augment the effects of UVR exposure. It has only been in the last quarter of a Century that it was found that UVR could cause immunologic changes that allow multiple skin cancers to develop. Advances in molecular biology have begun to show the cellular and molecular events that lead to UVR induced skin carcinogenesis. PMID- 11861224 TI - Targeting hepatocytes for drug and gene delivery: emerging novel approaches and applications. AB - The asialoglycoprotein receptor (ASGP-R) on mammalian hepatocytes provides a unique means for the development of liver-specific carriers, such as liposomes, recombinant lipoproteins, and polymers for drug or gene delivery to the liver, especially to hepatocytes. The abundant receptors on the cells specifically recognize ligands with terminal galactose or N-acetylgalactosamine residues, and endocytose the ligands for an intracellular degradation process. The use of its natural ligand, i.e. asialofetuin, or synthetic ligands with galactosylated or lactosylated residues, such as galactosylated cholesterol, glycolipids, or galactosylated polymers has achieved significant targeting efficacy to the liver. There are several examples of successful targeted therapy for acute liver injury with asialofetuin-labeled and vitamin E-associated liposomes or with a caspase inhibitor loaded in sugar-carrying polymer particles, as well as for the delivery of a new antiviral agent, 9-(2-phosphonylmethoxyethyl)adenine. Liposome-mediated gene delivery to the liver is more difficult than to other organs, such as to lungs. It is still in its infancy due to difficulties in solving general issues, such as the circulatory stability of liposome-DNA complexes, and lysosomal or endosomal degradation of plasmid DNA. In spite of these existing concerns, several new approaches offer some reason for optimism, for example; intravenous injection of asialofetuin- or galactosylated cholesterol-labeled cationic liposomes has led to high transgene expression in the liver. In addition, specific antisense oligonucleotides against woodchuck hepatitis viruses incorporated into sialoorosomucoid-poly-L-lysine significantly inhibited viral replication in the liver. Finally, galactosylated polymers are promising for gene delivery, but require further studies to verify their potential applications. PMID- 11861225 TI - Current perspectives on tinnitus. PMID- 11861226 TI - Plagiocephaly and head binding. PMID- 11861227 TI - Vitamin D deficiency in UK Asian families: activating a new concern. AB - Vitamin D has steroid hormonal effects which can produce clinical symptoms and signs unrelated to calcium homoeostasis. Its deficiency has been implicated as a risk factor for diabetes, ischaemic heart disease, and tuberculosis in Asians. In this review, the incidence, aetiology, prevention, and treatment of symptomatic vitamin D deficiency in childhood are considered. A renewed public health campaign is required in the UK to address the continuing problem of vitamin D deficiency in Asian families. PMID- 11861228 TI - An evaluation of the First Parent Health Visitor Scheme. AB - AIMS: To assess outcomes in families who received the First Parent Health Visitor Scheme (FPHVS), in comparison with families who received conventional ("generic") health visiting. METHODS: Retrospective data on 2113 families were collected during 1986-92 as part of National Health Service (NHS) service provision. Prospective data were collected during 1993-98 on 459 mothers and their children, with outcomes assessed at one year (93% follow up) and two years (80% follow up). RESULTS: There were no differences between the groups of mothers in self esteem, locus of control, or depression rates. The women who received the FPHVS were more likely to have changed partners, but they also had a wider support network than comparison women, and consulted their general practitioner (GP) less often. Breast feeding rates were higher in the FPHVS mothers, who also gave their infants more fruit juice drinks than the comparison group. No differences were apparent in developmental outcome using the Bayley Scales at 1 and 2 years of age. Both height and weight Z scores at 2 years of age were lower in the FPHVS children than the comparison children. Receipt of the FPHVS was associated with increased use of electric socket covers and lower accident rates in the second year of life. No differences were seen in immunisation rates, uptake of child health surveillance, or use of hospital services. A higher proportion of families who received the FPHVS were registered on the local child protection register compared with comparison families. CONCLUSION: Clustering effects dominated the analysis, but overall this evaluation could not show a clear advantage for the FPHVS over conventional health visiting. PMID- 11861229 TI - Evaluation of a mental health outreach service for homeless families. AB - AIMS: To describe the characteristics of homeless children and families seen by the mental health outreach service (MHOS), to evaluate the impact of this service on the short term psychosocial functioning of children and parents, and to establish perceptions of, and satisfaction with, the service. METHODS: Twenty seven children from 23 families who were in receipt of the MHOS and 27 children from 23 families residing in other hostels where no such service was available were studied. The MHOS was delivered by a clinical nurse specialist with expertise in child mental health, who offered the following interventions: assessment and brief treatment of mental health disorders in children; liaison with agencies; and training of homeless centre staff. RESULTS: Children in the experimental group had a significantly higher decrease in Strengths and Difficulties Questionnaire (SDQ) total scores. Having received the intervention was the strongest predictor of improvement in SDQ total scores. There was no significant impact on parental mental health (General Health Questionnaire) scores. Homeless families and staff expressed high satisfaction with the MHOS. CONCLUSION: This MHOS for homeless families is an innovative intervention which meets the complex and multiple needs of a vulnerable population unable to access mainstream mental health services. The primary objective of the service was to improve child mental health problems; however, the service developed in a responsive way by meeting social and practical needs of families in addition to its clinical role. PMID- 11861230 TI - Somatic and psychological problems in a cohort of sexually abused boys: a six year follow up case-control study. AB - AIMS: To obtain information about the health and well being of 108 boys six years after their involvement with the same paedophile. METHODS: Case-control study of the health records of 93 male victims of a major episode of school based child sexual abuse and 93 matched controls. Interviews with a sample of their general practitioners. RESULTS: The number and frequency of reported health problems were similar in both cases and controls. However, abused boys were more likely than controls to present with symptoms that persisted for more than a year (31 cases compared with 10 controls). CONCLUSIONS: Boys who have previously suffered sexual abuse at school did not utilise primary health care services more than a group of age matched controls. They did not present with psychological or somatic problems different from those presented by non-abused boys. However, abused boys were more likely to complain of persistent somatic or psychological problems lasting more than a year. This pattern appeared to persist after the abuse had stopped and the perpetrator imprisoned. PMID- 11861231 TI - Head injuries in infants: the risks of bouncy chairs and car seats. AB - AIMS: To establish whether the incorrect use of bouncy chairs (BC) and car seats (CS) is a significant cause of injuries in babies. METHODS: Prospective study over a 12 month period. All babies under the age of 1 year, presenting to the department with a head injury were eligible. RESULTS: A total of 131 cases were recorded, 17 (13%) of which were associated with either BC or CS, the second largest aetiological group. All BC cases (11/11, 100%) were caused by a fall from a surface with the baby in the chair. In the CS group, 2/6 (33%) were caused by such a fall. The carer was aware in only one of the 13 cases involving a fall from a surface that there was any chance of injury from this mechanism. CONCLUSION: Falls from inappropriately placed bouncy chairs or car seats form a significant proportion of head injuries in infants, resulting in unnecessary and preventable morbidity. PMID- 11861232 TI - Should we advise parents to administer over the counter cough medicines for acute cough? Systematic review of randomised controlled trials. AB - AIMS: To determine the effectiveness of over the counter (OTC) cough medicines for acute cough in children. METHODS: Systematic review of randomised controlled trials (RCTs). An all language search of the Cochrane Acute Respiratory Infections Group specialised register, Cochrane Controlled Trials Register, Medline, Embase, and the UK Department of Health National Research Register was performed. RCTs comparing oral OTC cough preparations with placebo in children suffering from acute cough as a result of upper respiratory tract infection (URTI) in ambulatory settings, using cough symptoms as an outcome, were included. RESULTS: Six trials involving 438 children met all inclusion criteria. Antitussives, antihistamine-decongestant combinations, other fixed drug combinations, and antihistamines were no more effective than placebo in relieving symptoms of acute cough. Based on a single study, the mucolytic preparation letosteine was superior to placebo, with differences in cough scores ranging from 0.1 to 0.3 points from day 4 to day 10. Most drugs appeared to be well tolerated with a low incidence of mostly minor adverse effects. CONCLUSION: OTC cough medicines do not appear more effective than placebo in relieving symptoms of acute cough. Even if statistically significant, effect sizes were small and of doubtful clinical relevance. The number of trials in each category was small, and the results of this systematic review have to be interpreted with caution. Based on the available evidence from a small number of studies, we cannot recommend OTC cough medicines as a first line treatment for children with acute cough. PMID- 11861234 TI - Inhaler devices for asthma: do we follow the guidelines? AB - BACKGROUND: Despite national guidelines for asthma treatment, many children have troublesome symptoms. AIM: To assess the extent to which the use of inappropriate inhaler devices contributes to this problem. METHODS: Of 14 813 questionnaires distributed to schoolchildren, 6996 (47%) were returned identifying 1444 children using asthma inhalers. Inhalers were categorised as age appropriate or inappropriate according to national guidelines and were compared with those used by 75 patients attending a hospital clinic. RESULTS: A total of 35% of "schools" and 4% of "clinic" children reported using an inappropriate inhaler device. Most were using metered dose inhalers alone. Twenty four per cent of "schools" children < or = 5 years old did not use a spacer. Both children and parents overestimated the child's ability to use their inhaler. CONCLUSIONS: Large numbers of children are given inhalers they cannot use. To improve asthma care we must ensure that prescriptions reflect the age and ability of the child. Recent recommendations by the Department of Health in England and Wales stress the importance of seamless care between primary and secondary services. As the management of childhood asthma is guided primarily by secondary care providers, it is therefore imperative that general paediatricians know the difficulties and issues which are occurring in the community. This will enable them to lead and support necessary change. PMID- 11861236 TI - Poverty and Bugatti. PMID- 11861235 TI - Maternal compliance with nutritional recommendations in an allergy preventive programme. AB - AIMS: To assess maternal compliance with nutritional recommendations in an allergy preventive programme, and identify factors influencing compliance behaviour. METHODS: Randomised double-blind intervention study on the effect of infant formulas with reduced allergenicity in healthy, term newborns at risk of atopy. Maternal compliance with dietary recommendations concerning milk and solid food feeding was categorised. RESULTS: A total of 2252 newborns were randomised to one of four study formulas. The drop out rate during the first year of life was 13.5% (n = 304). The rates of high, medium, and low compliance to milk feeding during weeks 1-16 were 83.4%, 4.0%, and 7.5%; the corresponding rates to solid food feeding during weeks 1-24 were 60.0%, 12.1%, and 22.9%. In 5.1% of subjects no nutritional information was available. Low compliance was more frequent among non-German parents, parents with a low level of education, young mothers, smoking mothers, and those who weaned their infant before the age of 2 months. CONCLUSIONS: Evaluation of allergy preventive programmes should take into account non-compliance for assessing the preventive effectiveness on study outcome. PMID- 11861237 TI - The importance of testing for adrenoleucodystrophy in males with idiopathic Addison's disease. AB - X linked adrenoleucodystrophy (X-ALD) is considered to be a rare cause of Addison's disease, although several small series suggest a high incidence in young Addisonian males. A survey in the south west of England identified 12 male patients diagnosed with Addison's disease in the period 1987-99. In 10 of these (83%) X-ALD was the underlying cause; the other two were of autoimmune aetiology. Five boys had developed Addison's disease subsequent to the diagnosis of X-ALD. Of the remaining five, in three boys the diagnosis of X-ALD was considerably delayed (by six months to two years from that of Addison's disease) and in two it was only made as a result of this survey. We also identified a patient who presented with Addison's disease at the age of 5 years but was only diagnosed as having X-ALD at the age of 34 years; in the interim his diagnosis of adrenomyeloneuropathy had been missed. Our experience highlights the absolute necessity of measuring very long chain fatty acids in all males with idiopathic Addison's disease. PMID- 11861239 TI - Serum hyaluronic acid concentrations are increased in cystic fibrosis patients with liver disease. AB - AIM: To determine whether serum hyaluronic acid (HA) concentrations are abnormal in patients with cystic fibrosis (CF) liver disease, and if so, whether the abnormality is associated with disease severity. METHODS: A total of 74 patients with CF were assessed for evidence of liver involvement as indicated by clinical, ultrasound, and biochemical findings. Serum hyaluronic acid concentrations were measured and compared with concentrations in 293 normal controls. Lung function in the CF patients was also recorded. RESULTS: Thirty four CF patients had no evidence of liver disease; in these, serum HA concentrations were similar to those in healthy controls (median (range): 16.1 (9.4-75.1) v 15 (1-77) microg/l). Nineteen CF patients had established liver disease detected by clinical and ultrasound examination, with significantly increased HA concentrations (56.1 (26 355) microg/l). Serum HA concentrations were also significantly increased, although to a lesser extent, in 21 CF patients with an abnormal liver ultrasound scan alone (22.4 (9.5-43.4) microg/l). There was no correlation between serum HA concentration and lung function. CONCLUSION: Serum HA concentrations were significantly increased in children with clinical or ultrasound evidence of liver disease, being higher in those with more advanced hepatic damage. Despite the inflammation and fibrosis present in CF lungs there was no correlation between HA concentration and lung function, suggesting that high concentrations were a failure of hepatic clearance rather than overproduction in the lung. Longitudinal measurement of HA concentrations may prove a useful marker for the development of significant liver damage in CF patients. PMID- 11861240 TI - Risk adjusted mortality of critical illness in a defined geographical region. AB - AIMS: To evaluate the performance of the Paediatric Risk of Mortality (PRISM) score in a population of UK children and to use this score to examine severity of illness adjusted mortality of critically ill children <16 years old in a defined geographical region. METHODS: Observational study of a defined population of critically ill children (<16 years old) admitted to hospitals in the South West Region between 1 December 1996 and 30 November 1998. RESULTS: Data were collected from 1148 eligible admissions. PRISM was found to perform acceptably in this population. There was no significant difference between the overall number of observed deaths and those predicted by PRISM. Admissions with mortality risk 30% or greater had significantly greater odds ratio for death in general intensive care units compared with the tertiary paediatric intensive care unit. CONCLUSIONS: Children with a high initial risk of mortality based on PRISM score were significantly more likely to survive in a tertiary paediatric intensive care unit than in general intensive care units in this region. However, there was no evidence from this study that admissions with lower mortality risk than 30% had significantly worse mortality in non-tertiary general units than in tertiary paediatric intensive care units. PMID- 11861241 TI - Transient lymphopenia in acutely unwell young infants. AB - The clinical outcome of 42 acutely unwell infants <3 months old with lymphopenia was retrospectively compared with that of 42 controls. Lymphopenic infants were significantly more likely to require active resuscitation and intensive care, independent of total leucocyte count, gender, degree of prematurity, and diagnosis. PMID- 11861242 TI - Bronchodilator therapy and hyperactivity in preschool children. AB - The common report of parents of asthmatic children that inhaled/nebulised salbutamol causes overactive behaviour was investigated. Nineteen children were assessed in a standardised setting before and after the administration of nebulised salbutamol and placebo. Neither parental report nor observer ratings suggested any significant increase in the child's level of activity. PMID- 11861244 TI - Cerebral venous thrombosis during diabetic ketoacidosis. AB - Neurological deterioration during an episode of diabetic ketoacidosis is usually assumed to be caused by cerebral oedema. We present a case of cerebral venous sinus thrombosis presenting in a similar manner, also associated with severe iron deficiency anaemia. Computed tomography scanning provided the correct diagnosis and allowed institution of anticoagulation with improvement in neurological outcome. Neuroimaging should always be performed in suspected cerebral oedema associated with diabetic ketoacidosis in order to exclude other pathologies. PMID- 11861245 TI - "Spina ventosa"--tuberculous dactylitis. PMID- 11861246 TI - Type 2 diabetes in obese white children. AB - We report four white adolescents aged 13 to 15 years (three females, one male) from the south and west region of England who presented with type 2 diabetes mellitus associated with significant obesity (body mass index more than +3SDS) in the past two years. Although these are the first reported obese, white cases from the UK to present with diabetes, we believe this clinical scenario will become more prevalent given the epidemic of childhood obesity in this country. PMID- 11861247 TI - Fulminant pulmonary oedema after administration of a balanced electrolyte polyethylene glycol solution. PMID- 11861248 TI - Diabetes unmasked by electric shock. AB - Following an accidental electric shock, a boy with no previous symptoms developed hyperglycaemia, rapidly evolving into diabetes. He was aglycosuric for 24 hours after the shock. Islet cell antibodies were shown shortly after the accident. Although destined to develop diabetes, it seems likely that the electric shock unmasked impaired glucose tolerance. Glucose homoeostasis should be monitored in children following significant electric shocks. PMID- 11861250 TI - Post-thrombotic syndrome. AB - Post-thrombotic syndrome (PTS) is a potentially disabling complication occurring in up to 67% of adult patients following deep venous thrombosis (DVT). PTS has recently been recognised in children. We present three cases of symptomatic PTS in children, which occurred following the use of central venous lines (CVLs). In two cases, no symptoms of acute thrombosis were noted. The cases highlight the clinical presentation of this syndrome. A review of the literature revealed two reports describing PTS occurring in children following DVT with an estimated incidence of 7-12%. It is concluded that PTS is an important complication of DVT in children. The clinical findings of pain, swelling, and brawny induration are similar to adult patients. The effect on growing limbs is not known. Paediatricians should be aware of the potential of PTS in all children who are at risk of DVT, including patients with malignancy, congenital heart disease, and children who have had previous CVLs, even in the absence of documented acute DVT. PMID- 11861251 TI - Investigation for complement deficiency following meningococcal disease. AB - BACKGROUND AND AIMS: The incidence of complement abnormalities in the UK is not known. It is suggested in at least three major paediatric textbooks to test for abnormalities of the complement system following meningococcal disease (MCD). METHODS: Over a four year period, surviving children with a diagnosis of MCD had complement activity assessed. A total of 297 children, aged 2 months to 16 years were screened. RESULTS: All children except one had disease caused by B or C serogroups. One child, with group B meningococcal septicaemia (complicated by disseminated intravascular coagulation and who required ventilation and inotropic support) was complement deficient. C2 deficiency was subsequently diagnosed. She had other major pointers towards an immunological abnormality prior to her MCD. CONCLUSION: It is unnecessary to screen all children routinely following MCD if caused by group B or C infection. However, it is important to assess the previous health of the child and to investigate appropriately if there have been previous suspicious infections, abnormal course of infective illnesses, or if this is a repeated episode of neisserial infection. PMID- 11861252 TI - Plasma and urinary soluble adhesion molecule expression is increased during first documented acute pyelonephritis. AB - BACKGROUND: The degree of inflammatory reaction and leucocyte trafficking during acute pyelonephritis has been related to the risk of developing renal parenchymal scarring. Adhesion molecules play a central role in leucocyte recruitment during inflammation. AIMS: (1) To determine whether circulating and urinary concentrations of E-selectin and intercellular adhesion molecule 1 (ICAM-1) were abnormal during first documented acute pyelonephritis; (2) to investigate whether circulating or urinary concentrations were predictive for the development of abnormalities on DMSA imaging. METHODS: Plasma and urine samples were collected from 40 children with a first episode of acute pyelonephritis within one week of infection (acute sample) and at six weeks (late sample). Control samples were collected from 21 healthy age matched controls and 18 age matched controls with febrile illness not secondary to urinary tract infection. RESULTS: Plasma and urinary sE-selectin were higher in acute samples (median 176.3 ng/ml and 0.12 ng/mmol respectively) compared with late (97.8 ng/ml and 0.029 ng/mmol) and both control (65.6 ng/ml and 0 ng/mmol) and febrile control (urine 0 ng/mmol) samples. Plasma sICAM-1 was higher in acute samples (428 ng/ml) than controls (365.2 ng/ml), and acute sICAM-1 urine concentrations were higher than febrile control concentrations (3.2 v 0.7 ng/mmol). No correlations were detected between sE selectin or sICAM-1 and acute or late DMSA scan changes. CONCLUSION: Plasma and urinary sE-selectin and sICAM-1 are significantly increased during acute pyelonephritis, though no correlation exists between the presence of high plasma or urine concentrations and DMSA scan changes, both during acute infection and six weeks post-infection. PMID- 11861253 TI - A diatribe on dummies. PMID- 11861254 TI - Information technology--improving medicine. PMID- 11861255 TI - Kocher Debre Semelaigne syndrome: regression of pesudohypertrophy of muscles on thyroxine. PMID- 11861256 TI - Break dancer's lung. PMID- 11861258 TI - Sugar in erythropoietin: clinical and forensic implications. PMID- 11861259 TI - "Special delivery" to cancer cells. PMID- 11861260 TI - Defining the genetic chaos in myeloma. PMID- 11861261 TI - Frequent expression of CCR4 in adult T-cell leukemia and human T-cell leukemia virus type 1-transformed T cells. AB - Chemokines and chemokine receptors play important roles in migration and tissue localization of various lymphocyte subsets. Here, we report the highly frequent expression of CCR4 in adult T-cell leukemia (ATL) and human T-cell leukemia virus type 1 (HTLV-1)-immortalized T cells. Flow cytometric analysis revealed that ATL and HTLV-1-immortalized T-cell lines consistently expressed CCR4. Inducible expression of HTLV-1 transcriptional activator tax in a human T-cell line Jurkat did not, however, up-regulate CCR4 mRNA. In vitro immortalization of peripheral blood T cells led to preferential outgrowth of CD4(+) T cells expressing CCR4. We further demonstrated highly frequent expression of CCR4 in fresh ATL cells by (1) reverse transcriptase-polymerase chain reaction (RT-PCR) analysis of CCR4 expression in peripheral blood mononuclear cells (PBMCs) from patients with ATL and healthy controls; (2) flow cytometric analysis of CCR4-expressing cells in PBMCs from patients with ATL and healthy controls; (3) CCR4 staining of routine blood smears from patients with ATL; and (4) an efficient migration of fresh ATL cells to the CCR4 ligands, TARC/CCL17 and MDC/CCL22, in chemotaxis assays. Furthermore, we detected strong signals for CCR4, TARC, and MDC in ATL skin lesions by RT-PCR. Collectively, most ATL cases have apparently derived from CD4(+) T cells expressing CCR4. It is now known that circulating CCR4(+) T cells are mostly polarized to Th2 and also contain essentially all skin-seeking memory T cells. Thus, HTLV-1-infected CCR4(+) T cells may have growth advantages by deviating host immune responses to Th2. CCR4 expression may also account for frequent infiltration of ATL into tissues such as skin and lymph nodes. PMID- 11861262 TI - Inhibition of macrophage inflammatory protein-1 alpha production by Epstein-Barr virus. AB - Infection with Epstein-Barr virus (EBV) exerts substantially immunomodulating activities in vitro and in vivo. In this context, EBV-induced chemokine production and the influence of EBV on this highly redundant system of inflammatory proteins have hardly been investigated. This study analyzed the production of interleukin-8, RANTES, monocyte chemotactic protein-1, and macrophage inflammatory protein-1 alpha (MIP-1 alpha) on EBV infection of peripheral blood mononuclear cells from immune EBV-seropositive (EBV(+)) and noninfected EBV-seronegative (EBV(-)) individuals. EBV failed to induce the production of MIP-1 alpha in EBV(+) as well as EBV(-) individuals, whereas the other chemokines studied were readily expressed. Moreover, EBV completely down regulated lipopolysaccharide (LPS)- and phytohemagglutinin-induced MIP-1 alpha production up to 4 hours after induction. Reverse transcription-polymerase chain reaction (RT-PCR) analysis of EBV- and LPS-stimulated cultures revealed that EBV inhibited MIP-1 alpha production on the transcriptional level. This effect was abolished by addition of antiglycoprotein (gp)350/220, a monoclonal antibody against EBV's major envelope glycoprotein, which mediates binding of the virus to the EBV receptor, CD21. However, recombinant gp350/220 protein alone did not inhibit the LPS-induced MIP-1 alpha production, indicating that infection of the target cell is indispensable for this effect. In summary, we demonstrate a new immunomodulating activity of EBV on the chemokine system that probably helps the virus to evade the host's immune system favoring lifelong infection. PMID- 11861263 TI - Idiotype-pulsed dendritic cell vaccination for B-cell lymphoma: clinical and immune responses in 35 patients. AB - Tumor-specific clonal immunoglobulin expressed by B-cell lymphomas (idiotype [Id]) can serve as a target for active immunotherapy. We have previously described the vaccination of 4 patients with follicular lymphoma using dendritic cells (DCs) pulsed with tumor-derived Id protein and now report on 35 patients treated using this approach. Among 10 initial patients with measurable lymphoma, 8 mounted T-cell proliferative anti-Id responses, and 4 had clinical responses--2 complete responses (CRs) (progression-free [PF] for 44 and 57 months after vaccination), 1 partial response (PR) (PF for 12 months), and 1 molecular response (PF for 75+ months). Subsequently, 25 additional patients were vaccinated after first chemotherapy, and 15 of 23 (65%) who completed the vaccination schedule mounted T-cell or humoral anti-Id responses. Induction of high-titer immunoglobulin G anti-Id antibodies required coupling of Id to the immunogenic carrier protein keyhole limpet hemocyanin (Id-KLH). These antibodies could bind to and induce tyrosine phosphorylation in autologous tumor cells. Among 18 patients with residual tumor at the time of vaccination, 4 (22%) had tumor regression, and 16 of 23 patients (70%) remain without tumor progression at a median of 43 months after chemotherapy. Six patients with disease progression after primary DC vaccination received booster injections of Id-KLH protein, and tumor regression was observed in 3 of them (2 CRs and 1 PR). We conclude that Id pulsed DC vaccination can induce T-cell and humoral anti-Id immune responses and durable tumor regression. Subsequent boosting with Id-KLH can lead to tumor regression despite apparent resistance to the primary DC vaccine. PMID- 11861264 TI - A randomized study of interferon-alpha versus interferon-alpha and low-dose arabinosyl cytosine in chronic myeloid leukemia. AB - Interferon-alpha (IFN-alpha) has significantly prolonged survival in chronic myeloid leukemia (CML), but some patients do not respond and many responses are not durable. To improve the results, IFN-alpha has been combined with other treatments, but so far only the association with low-dose arabinosyl cytosine (LDAC) has been shown to increase the response rate and to prolong survival. Here are reported the results of a study of 538 Philadelphia chromosome-positive CML patients who were assigned at random to treatment with IFN-alpha 2a alone or in combination with LDAC. The scheduled dose of IFN-alpha 2a was 5(6) IU/m(2)/d. The scheduled dose of AC was 40 mg/d for the first 10 days of each month of treatment. The efficacy endpoints were a complete hematologic response rate at 6 months (62% in the IFN-alpha-plus-LDAC arm versus 55% in the IFN-alpha arm; P =.11), major cytogenetic response (MCgR) rate at 24 months (28% versus 18%; P =.003), and overall survival (5-year survival, 68% versus 65%; P =.77). Treatment did not affect overall survival within different prognostic risk groups: low, intermediate, or high. Also the duration of MCgR was identical. The results of this study confirm the results of a similar French study only for the response rate, not for survival, suggesting that the relationship between cytogenetic response and survival may be extremely variable and that a meta-analysis of these and other studies of IFN-alpha versus IFN-alpha plus LDAC is required to settle the issue of the role of LDAC in the treatment of CML. PMID- 11861265 TI - Leading prognostic relevance of the BCR-ABL translocation in adult acute B lineage lymphoblastic leukemia: a prospective study of the German Multicenter Trial Group and confirmed polymerase chain reaction analysis. AB - The BCR-ABL fusion, the molecular equivalent of the Philadelphia translocation, gains importance for treatment stratification in adult acute lymphoblastic leukemia (ALL). In this prospective study, samples from 478 patients with CD10(+) B-cell precursor ALL (c-ALL and pre-B ALL) underwent BCR-ABL reverse transcription-polymerase chain reaction (RT-PCR) analysis with double testing of positive samples. Patients were stratified according to the PCR result and treated in 2 German Multicenter Trials of Adult ALL. The outcome was followed and the prognostic impact of BCR-ABL was compared to clinical risk features. Of the 478 samples, 432 had an evaluable BCR-ABL result. Thirty-seven percent of the c ALL and pre-B ALL patients were BCR-ABL(+) (p190, 77%; p210, 20%; simultaneous p190/p210, 3%). BCR-ABL positivity was associated with the high-risk features of older age (45 years versus 30 years median age; P =.0001) and higher white blood cell counts (23 500/microL versus 11 550/microL; P =.0001). Univariate and multivariate analyses revealed BCR-ABL as the leading factor for a poor prognosis (P =.0001) in comparison to clinical risk criteria. Irrespective of the breakpoint, presence of any BCR-ABL transcript predicted a lower chance of initial treatment response (68.4% versus 84.6%; P =.001) and a lower probability of disease-free survival at 3 years (0.13 versus 0.47; P =.0001). This bad outcome was not influenced by postinduction high-dose treatment stratifications. The results show a high prevalence of BCR-ABL fusion transcripts with predominance of p190. BCR-ABL RT-PCR is confirmed as a sensitive, rapid method to diagnose t(9;22), and p190 and p210 are unequivocally demonstrated as the most important predictors of poor long-term survival despite intensified chemotherapy. PMID- 11861266 TI - Severe deficiency of switched memory B cells (CD27(+)IgM(-)IgD(-)) in subgroups of patients with common variable immunodeficiency: a new approach to classify a heterogeneous disease. AB - Hypogammaglobulinemia is the hallmark of common variable immunodeficiency (CVID) syndrome, a heterogeneous disorder predisposing patients to recurrent bacterial infections. In this study, we investigated the peripheral B-cell compartment of 30 well-characterized CVID patients in comparison to 22 healthy controls. Flow cytometric analysis of peripheral blood lymphocytes revealed a reduction of class switched CD27(+)IgM(-)IgD(-) memory B cells below 0.4% in 77% of our patients (group I), while this B-cell subpopulation exceeded 0.5% in all healthy donors and in 23% of CVID patients (group II). These results correlate well with the capacity of peripheral blood lymphocytes to produce immunoglobulins in vitro upon stimulation with Staphylococcus aureus Cowan I (SAC) plus interleukin-2 because the production of immunoglobulin G in vitro is entirely dependent on the presence of switched memory B cells. The subdivision of group I into patients with an increased proportion of CD21(-) peripheral B cells (> 20%; group Ia) and patients with normal percentages of CD21(-) B cells (< 20%; group Ib) revealed a significant clustering of patients with splenomegaly and autoimmune cytopenias in group Ia. Based on these observations, we propose a fast and reliable new classification for CVID patients by flow cytometric quantification of class switched memory and immature B cells in the peripheral blood of patients. Our results point toward defects at various stages of B-cell differentiation in CVID subgroups and support the value of a B-cell-oriented classification principle. A consensus on this new classification system will hopefully provide a tool for rapidly defining homogeneous subgroups of CVID for functional studies and genetic linkage analysis. PMID- 11861267 TI - Effect of N-acetyl-cysteine on the hypoxic ventilatory response and erythropoietin production: linkage between plasma thiol redox state and O(2) chemosensitivity. AB - Oxygen-sensing chemoreceptors contribute significantly to the regulation of the respiratory drive and arterial PO(2) levels. The hypoxic ventilatory response (HVR) decreases strongly with age and is modulated by prolonged hypoxia and physical exercise. Several earlier studies indicated that the regulation of the ventilatory response and erythropoietin (EPO) production by the respective oxygen sensors involves redox-sensitive signaling pathways, which are triggered by the O(2)-dependent production of reactive oxygen species. The hypothesis that HVR and EPO production are modulated by thiol compounds or changes in the plasma thiol disulfide redox state (REDST) was investigated. It was demonstrated that both responses are enhanced by oral treatment with N-acetyl-cysteine (NAC) and that HVR is correlated with plasma thiol level and REDST. Results suggest the possibility that age-related changes in plasma REDST may account for the age related changes in HVR. PMID- 11861269 TI - Role of erythrocyte phosphatidylserine in sickle red cell-endothelial adhesion. AB - Phosphatidlyserine (PS) exposure on the erythrocyte surface endows the cell with the propensity of adhering to vascular endothelium. Because individuals with sickle cell disease (SCD) manifest loss of erythrocyte membrane asymmetry with PS exposure, we have assessed the contribution of this marker to the process of sickle erythrocyte-microendothelial adhesion. Assays for plasma-induced adhesion were conducted on unactivated endothelium, in the absence of immobilized ligands, such that PS was compared to the erythrocyte adhesion receptor CD36. Blocking studies with erythrocytes pretreated with annexin V (to cloak PS) or anti-CD36 or both revealed an inhibitory effect on adhesion of 36% +/- 10% and 23% +/- 8% with blocking of both sites suggestive of an additive effect. We next evaluated 87 blood samples from patients with SCD and grouped them into 4 categories based on adhesion marker (CD36 and PS) levels. Results revealed a striking correlation between erythrocyte PS positivity and adhesion. Analyses of the individual patient data demonstrated a positive correlation between PS and adhesion (R = 0.52, P <.000 001), whereas none was noted between adhesion and CD36 (R = 0.2, P >.07). The effect of PS on adhesion appears to be related to the quantitative differences in erythrocyte markers in SCD, with PS the predominant marker when compared to CD36 both in the total erythrocyte population, and when the adherence prone erythrocyte, the CD71(+) stress reticulocyte, was evaluated. Our study signals the entrance of an important new contributor to the field of sickle erythrocyte-endothelial adhesion. The implications of erythrocyte PS exposure in relation to the vascular pathology of SCD need to be assessed. PMID- 11861268 TI - Clinical and biologic features of CD4(+)CD56(+) malignancies. AB - CD4(+)CD56(+) malignancies are rare hematologic neoplasms, which were recently shown to correspond to the so-called type 2 dendritic cell (DC2) or plasmacytoid dendritic cells. This study presents the biologic and clinical features of a series of 23 such cases, selected on the minimal immunophenotypic criteria defining the DC2 leukemic counterpart, that is, coexpression of CD4 and CD56 in the absence of B, T, and myeloid lineage markers. Clinical presentation typically corresponded to cutaneous nodules associated with lymphadenopathy or spleen enlargement or both. Cytopenia was frequent. Circulating malignant cells were often detected. Massive bone marrow infiltration was seen in 20 of 23 (87%) patients. Most tumor cells exhibited nuclei with a lacy chromatin, a blastic aspect, large cytoplasm-containing vacuoles or microvacuoles beside the plasma membrane, and cytoplasmic expansions resembling pseudopodia. Other immunophenotypic characteristics included both negative (CD16, CD57, CD116, and CD117) and positive (CD36, CD38, CD45 at low levels, CD45RA, CD68, CD123, and HLA DR) markers. The prognosis was rapidly fatal in the absence of chemotherapy. Complete remission was obtained in 18 of 23 (78%) patients after polychemotherapy. Most patients had a relapse in less than 2 years, mainly in the bone marrow, skin, or central nervous system. Considering these clinical and biologic features, the conclusion is made that CD4(+)CD56(+) malignancies constitute a genuine homogeneous entity. Furthermore, some therapeutic options were clearly identified. Finally, relationships between the pure cutaneous indolent form of the disease and acute leukemia as well as with the lymphoid/myeloid origin of the CD4(+)CD56(+) malignant cell are discussed. PMID- 11861271 TI - Sustained response to recombinant human erythropoietin and intermittent all-trans retinoic acid in patients with myelodysplastic syndromes. AB - In vitro studies suggest that all-trans retinoic acid (ATRA) synergizes with erythropoietin (EPO) for the stimulation of hematopoiesis in patients with myelodysplastic syndrome (MDS). A clinical trial was performed to evaluate whether a combination of these agents was effective in relieving the cytopenias associated with MDS. Twenty-seven patients with low- or intermediate-risk MDS were enrolled in a 12-week study. ATRA was administered orally at the dose of 80 mg/m(2) per day in 2 divided doses for 7 consecutive days every other week. Recombinant human EPO was given subcutaneously 3 times a week. The EPO dose was initiated at 150 U/kg and was increased to 300 U/kg if after 6 weeks there was no or there was suboptimal erythroid response. Patients who responded to therapy were continued on ATRA and EPO at the same doses for 6 additional months (extension phase). Further treatment was given to patients with a continued response. Clinically significant erythroid responses with increases of hemoglobin levels of at least 1 g/dL or reduction of transfusion needs were seen in 13 (48%) patients, with 4 showing improved responses after dose escalation of EPO. Ten (37%) patients displayed continued responses during 6 months of extended treatment, and 7 (26%) are still responsive after a follow-up period of 13 months. Neutrophil responses were observed in 5 of 12 patients with neutropenia, and platelet responses were observed in 6 of 9 patients with thrombocytopenia. Three patients displayed trilineage responses that were sustained during continuation therapy. Side effects were observed in all patients but were of mild entity and did not require discontinuation of therapy. It is concluded that the combination ATRA + EPO is an effective and well-tolerated treatment for patients with low- and intermediate-risk MDS. The optimal ATRA and EPO schedule and the role of maintenance treatment remain to be determined and warrant further investigation. PMID- 11861270 TI - Effect of tolerance to noninherited maternal antigens on the occurrence of graft versus-host disease after bone marrow transplantation from a parent or an HLA haploidentical sibling. AB - In haploidentical transplantation, the mismatched haplotype of the donor can originate from either of the parents. We refer to such mismatched haplotypes as noninherited maternal antigens (NIMA haplotype) or noninherited paternal antigens (NIPA haplotype). To determine whether exposure to maternal HLA antigens benefits patients undergoing bone marrow transplantation, we analyzed graft failure and graft-versus-host disease (GVHD) after transplantations from parental or haploidentical sibling donors. We studied 269 patients receiving 1 or 2 HLA-A, B, -DR antigen-mismatched sibling or parental non-T-cell-depleted bone marrow transplants for acute myelogenous leukemia, acute lymphoblastic leukemia, or chronic myelogenous leukemia between 1985 and 1997 that were reported to the International Bone Marrow Transplant Registry. Included were 121 (45%) NIMA mismatched and 148 (55%) NIPA-mismatched transplantations. Sixty-three (52%) of the NIMA-mismatched transplants and 69 (47%) of the NIPA-mismatched transplants were from haploidentical sibling donors. Sibling transplantations mismatched for NIMA had similar rates of graft failure but lower rates of acute GVHD (P <.02) than NIPA-mismatched sibling transplantations. In the first 4 months after transplantation, mother-to-child transplantations involved significantly less chronic GVHD than father-to-child transplantations (P <.02). Treatment-related mortality (TRM) was significantly higher after parental transplantations (P =.009 for mother; P =.03 for father) than after haploidentical sibling transplantations mismatched for the NIMA. Non-T-cell-depleted bone marrow transplants donated by haploidentical siblings to recipients mismatched for NIPA and transplants donated by parents caused more acute and chronic GVHD and TRM than transplants donated by haploidentical siblings mismatched for NIMA. PMID- 11861272 TI - Homing efficiency, cell cycle kinetics, and survival of quiescent and cycling human CD34(+) cells transplanted into conditioned NOD/SCID recipients. AB - Differences in engraftment potential of hematopoietic stem cells (HSCs) in distinct phases of cell cycle may result from the inability of cycling cells to home to the bone marrow (BM) and may be influenced by the rate of entry of BM homed HSCs into cell cycle. Alternatively, preferential apoptosis of cycling cells may contribute to their low engraftment potential. This study examined homing, cell cycle progression, and survival of human hematopoietic cells transplanted into nonobese diabetic severe combined immunodeficient (NOD/SCID) recipients. At 40 hours after transplantation (AT), only 1% of CD34(+) cells, or their G(0) (G(0)CD34(+)) or G(1) (G(1)CD34(+)) subfractions, was detected in the BM of recipient mice, suggesting that homing of engrafting cells to the BM was not specific. BM of NOD/SCID mice receiving grafts containing approximately 50% CD34(+) cells harbored similar numbers of CD34(+) and CD34(-) cells, indicating that CD34(+) cells did not preferentially traffic to the BM. Although more than 64% of human hematopoietic cells cycled in culture at 40 hours, more than 92% of cells recovered from NOD/SCID marrow were quiescent. Interestingly, more apoptotic human cells were detected at 40 hours AT in the BM of mice that received xenografts of expanded cells in S/G(2)+M than in recipients of G(0)/G(1) cells (34.6% +/- 5.9% and 17.1% +/- 6.3%, respectively; P <.01). These results suggest that active proliferation inhibition in the BM of irradiated recipients maintains mitotic quiescence of transplanted HSCs early AT and may trigger apoptosis of cycling cells. These data also illustrate that trafficking of transplanted cells to the BM is not selective, but lodgment of BM-homed cells may be specific. PMID- 11861273 TI - In vitro proliferation and differentiation of erythroid progenitors from patients with myelodysplastic syndromes: evidence for Fas-dependent apoptosis. AB - Erythropoiesis results from the proliferation and differentiation of pluripotent stem cells into immature erythroid progenitors (ie, erythroid burst-forming units (BFU-Es), whose growth, survival, and terminal differentiation depends on erythropoietin (Epo). Ineffective erythropoiesis is a common feature of myelodysplastic syndromes (MDS). We used a 2-step liquid-culture procedure to study erythropoiesis in MDS. CD34(+) cells from the marrow of patients with MDS were cultured for 10 days in serum-containing medium with Epo, stem cell factor, insulin-like growth factor 1, and steroid hormones until they reached the proerythroblast stage. The cells were then placed in medium containing Epo and insulin for terminal erythroid differentiation. Numbers of both MDS and normal control cells increased 10(3) fold by day 15. However, in semisolid culture, cells from patients with refractory anemia (RA) with ringed sideroblasts and RA or RA with excess of blasts produced significantly fewer BFU-Es than cells from controls. Fluorescence in situ hybridization analysis of interphase nuclei from patients with chromosomal defects indicated that abnormal clones were expanded in vitro. Epo-signaling pathways (STAT5, Akt, and ERK 1/2) were normally activated in MDS erythroid progenitors. In contrast, apoptosis was significantly increased in MDS cells once they differentiated, whereas it remained low in normal cells. Fas was overexpressed on freshly isolated MDS CD34(+) cells and on MDS erythroid cells throughout the culture. Apoptosis coincided with overproduction of Fas ligand during the differentiation stage and was inhibited by Fas-Fc chimeric protein. Thus, MDS CD34(+)-derived erythroid progenitors proliferated normally in our 2-step liquid culture with Epo but underwent abnormal Fas-dependent apoptosis during differentiation that could be responsible for the impaired erythropoiesis. PMID- 11861274 TI - Effects of anagrelide on in vivo megakaryocyte proliferation and maturation in essential thrombocythemia. AB - To define the mechanism by which anagrelide normalizes the platelet count in essential thrombocythemia, we studied in vivo megakaryocytopoiesis in 10 newly diagnosed patients prior to and while on anagrelide therapy. Using flow cytometric analysis of aspirated marrow, megakaryocytopoiesis was quantified and correlated with the autologous platelet production rate. Megakaryocytes were identified by CD41a expression and enumerated in relation to the nucleated marrow erythroid precursors. Megakaryocyte diameters were directly measured by time-of flight technique, and cell ploidy was measured by DNA staining. Two to 3 thousand megakaryocytes were analyzed in each sample. In the 10 patients, the platelet count was 1063 +/- 419 x 10(9) platelets/L (mean +/- 1 SD) with markedly increased production (237 +/- 74 x 10(9) platelets/L per day versus 43.1 +/- 8.4 x 10(9) platelets/L per day in healthy individuals). The platelet survival was 8.2 +/- 1.1 days versus 9.0 +/- 0.5 days in healthy controls (P >.05). Megakaryocyte diameter was increased to 46 microm (versus 37 microm in controls; range, 21 microm for 2N to 56 microm for 64N cells). The volume increased to 48 x 10(3) microm(3) versus 26 x 10(3) microm(3) in controls, and the number increased to 14 x 10(6)/kg (versus 7 x 10(6)/kg in controls), resulting in 3.7-fold increase in megakaryocyte mass (66 x 10(10) microm(3)/kg versus 18 x 10(10) microm(3)/kg). Cell ploidy was enhanced showing a modal ploidy of 32N (versus 16N in healthy controls) with marked increase in 64N and 128N cells (P <.05). Anagrelide therapy reduced the platelet counts to 361 +/- 53 x 10(9) platelets/L and the turnover rate to 81 x 10(9) platelets/L per day. The platelet survival was unchanged. Following therapy, megakaryocyte number decreased to 8 x 10(6)/kg, diameter to 40 microm, and volume to 34 x 10(3) microm(3) with a normalized modal ploidy of 16N, resulting in a megakaryocyte mass reduced by 60% (28 x 10(10) microm(3)/kg; P <.05). This reduction in cell mass closely correlated with the reduction in platelet count and production rate by 66% (r = 0.96). The present data indicate that in essential thrombocythemia anagrelide therapy decreases circulating platelets by reducing both megakaryocyte hyperproliferation and differentiation. PMID- 11861276 TI - Adapted NOD/SCID model supports development of phenotypically and functionally mature T cells from human umbilical cord blood CD34(+) cells. AB - The NOD-LtSZ scid/scid (NOD/SCID) repopulation assay is the criterion for the study of self-renewal and multilineage differentiation of human hematopoietic stem cells. An important shortcoming of this model is the reported absence of T cell development. We studied this aspect of the model and investigated how it could be optimized to support T-cell development. Occasionally, low-grade thymic engraftment was observed in NOD/SCID mice or Rag2(-/-)gamma(c)(-/-) mice. In contrast, the treatment of NOD/SCID mice with a monoclonal antibody against the murine interleukin-2R beta, (IL-2R beta) known to decrease natural killer cell activity, resulted in human thymopoiesis in up to 60% of the mice. T-cell development was phenotypically normal and resulted in polyclonal, mature, and functional CD1(-) TCR alpha beta (+) CD4(+) or CD8(+) single-positive T cells. In mice with ongoing thymopoiesis, peripheral T cells were observed. TREC analysis showed that T cells with a naive phenotype (CD45RA(+)) emerged from the thymus. In approximately half of these mice, the peripheral T cells included a pauciclonal outgrowth of CD45RO(+) cells. These data suggest that all elements of a functional immune system were present in these animals. PMID- 11861275 TI - Bone marrow progenitor cell reserve and function and stromal cell function are defective in rheumatoid arthritis: evidence for a tumor necrosis factor alpha mediated effect. AB - Based on previous reports for impaired hematopoiesis in rheumatoid arthritis (RA), and in view of the current interest in exploring the role of autologous stem cell transplantation (ASCT) as an alternative treatment in patients with resistant disease, we have evaluated bone marrow (BM) progenitor cell reserve and function and stromal cell function in 26 patients with active RA. BM progenitor cells were assessed using flow cytometry and clonogenic assays in short-term and long-term BM cultures (LTBMCs). BM stroma function was assessed by evaluating the capacity of preformed irradiated LTBMC stromal layers to support the growth of normal CD34(+) cells. We found that RA patients exhibited low number and increased apoptosis of CD34(+) cells, defective clonogenic potential of BM mononuclear and purified CD34(+) cells, and low progenitor cell recovery in LTBMCs, compared with healthy controls (n = 37). Patient LTBMC stromal layers failed to support normal hematopoiesis and produced abnormally high amounts of tumor necrosis factor alpha (TNF alpha). TNF alpha levels in LTBMC supernatants inversely correlated with the proportion of CD34(+) cells and the number of colony-forming cells, and positively with the percentage of apoptotic CD34(+) cells. Significant restoration of the disturbed hematopoiesis was obtained following anti-TNF alpha treatment in 12 patients studied. We concluded that BM progenitor cell reserve and function and BM stromal cell function are defective in RA probably due, at least in part, to a TNF alpha-mediated effect. The role of these abnormalities on stem cell harvesting and engraftment in RA patients undergoing ASCT remains to be clarified. PMID- 11861277 TI - Differential expression and phosphorylation of distinct STAT3 proteins during granulocytic differentiation. AB - External stimuli act in concert with intracellular signals to regulate a cell's genetic program, activating genes important in granulocytic lineage commitment, proliferation, and maturation. Signal transducer and activator of transcription 3 (STAT3), a transcription factor, has been implicated in mediating granulocytic differentiation. We have examined the role of STAT3 as a physiologic mediator of granulocytic kinetics. Distinct isoforms--the long form STAT3 alpha, the truncated forms STAT3 beta and STAT3 gamma, and a putative novel form STAT3 delta -were expressed and activated in a maturation stage-specific manner. With the progression of differentiation, the ratio of isoforms shifted from predominantly STAT3 alpha to STAT3 beta. The kinetics of STAT3 gamma, generated through proteolytic cleavage of STAT3 alpha, coincided with but were inverse to those of STAT3 alpha. STAT3 delta was expressed at low levels and decreased with differentiation but was preferentially phosphorylated during an intermediate stage of maturation. Under different culture conditions (pH, O(2) tension [pO(2)], IL-3), we found that the expression and phosphorylation status of the different STAT3 isoforms displayed unique kinetic patterns that correlated with the effects on granulocyte differentiation. The evidence suggests that signals triggered by pH, pO(2), and IL-3 each converge on STAT3 through independent mechanisms, exploiting the flexibility granted by the diversity in expression and phosphorylation of the different STAT3 isoforms, to regulate distinct granulocytic cell responses. The selective expression of STAT3 isoforms and their activation is a major determinant of granulocytic cell development and provides a molecular basis for evaluating the effects of various environmental factors on the STAT3-mediated signaling pathway. PMID- 11861278 TI - Antithrombin prevents endotoxin-induced hypotension by inhibiting the induction of nitric oxide synthase in rats. AB - Antithrombin (AT) prevents Escherichia coli-induced hypotension in animal models of sepsis, and it further reduces the mortality of patients with septic shock. In the present study, we examined whether AT may prevent the endotoxin (ET)-induced hypotension by promoting the endothelial release of prostacyclin (PGI(2)) in rats. Intravenous administration of AT (250 U/kg) prevented both hypotension and the increases in plasma levels of NO(2)(-)/NO(3)(-) in rats given ET. Lung expression of messenger RNA (mRNA) for tumor necrosis factor-alpha (TNF-alpha) was transiently increased after ET administration, followed by the increases in lung tissue levels of TNF-alpha. Both the lung activity of the inducible form of nitric oxide synthase (iNOS) and the lung expression of iNOS mRNA in animals administered ET were gradually increased after the TNF-alpha mRNA expression had peaked. Administration of AT significantly inhibited these increases. Neither DEGR-F.Xa, a selective inhibitor of thrombin generation, nor Trp(49)-modified AT, which is not capable of promoting the endothelial release of PGI(2), showed any effects on these changes induced by ET. Administration of antirat TNF-alpha antibody produced effects similar to those induced by AT. Indomethacin pretreatment abrogated the effects induced by AT. Iloprost, a stable derivative of PGI(2), produced effects similar to those of AT. These findings suggested that AT prevents the ET-induced hypotension by inhibiting the induction of iNOS through inhibiting TNF-alpha production. These effects of AT could be mediated by the promotion of endothelial release of PGI(2) and might at least partly explain the therapeutic effects for septic shock. PMID- 11861279 TI - Thrombin induces increased expression and secretion of angiopoietin-2 from human umbilical vein endothelial cells. AB - Angiogenesis is required for tumor growth and metastasis. It has recently been suggested that thrombin is a potent promoter of angiogenesis. We therefore examined the possibility that thrombin could be inducing the expression of angiopoietin-2 (Ang-2), necessary for remodeling. Human umbilical vein endothelial cells were incubated with or without thrombin (1 U/mL) for 1 to 24 hours and then examined for messenger RNA (mRNA) by Northern analysis. Enhanced mRNA expression (about 4-fold over baseline) was noted at 4 hours. Enhanced expression of Ang-2 mRNA was secondary to enhanced transcription (about 4-fold), with no effect on stabilization. Enhanced Ang-2 mRNA transcription was inhibited by H7 and PD98059, indicating the requirement of serine/threonine kinases as well as the mitogen-activated protein kinase pathway. Up-regulation of mRNA was associated with enhanced Ang-2 protein synthesis and secretion as assayed by immunoblot. Thrombin-induced secreted Ang-2 inhibited the binding of recombinant (35)S-Ang-1 to its Tie-2-Fc receptor, demonstrating functionality. Hirudin reversed this effect, demonstrating thrombin specificity. Thus, thrombin-induced tumorigenesis and metastasis is associated with enhanced Ang-2 protein synthesis and secretion via enhanced transcription of Ang-2. This could help explain how thrombin promotes angiogenesis. PMID- 11861280 TI - The Hermansky-Pudlak syndrome 1 (HPS1) and HPS2 genes independently contribute to the production and function of platelet dense granules, melanosomes, and lysosomes. AB - Hermansky-Pudlak syndrome (HPS) is an inherited hemorrhagic disease affecting the related subcellular organelles platelet dense granules, lysosomes, and melanosomes. The mouse genes for HPS, pale ear and pearl, orthologous to the human HPS1 and HPS2 (ADTB3A) genes, encode a novel protein of unknown function and the beta(3)A subunit of the AP-3 adaptor complex, respectively. To test for in vivo interactions between these genes in the production and function of intracellular organelles, mice doubly homozygous for the 2 mutant genes were produced by appropriate breeding. Cooperation between the 2 genes in melanosome production was evident in increased hypopigmentation of the coat together with dramatic quantitative and qualitative alterations of melanosomes of the retinal pigment epithelium and choroid of double mutant mice. Lysosomal and platelet dense granule abnormalities, including hyposecretion of lysosomal enzymes from kidneys and depression of serotonin concentrations of platelet dense granules were likewise more severe in double than single mutants. Also, lysosomal enzyme concentrations were significantly increased in lungs of double mutant mice. Interaction between the 2 genes was specific in that effects on organelles were confined to melanosomes, lysosomes, and platelet dense granules. Together, the evidence indicates these 2 HPS genes function largely independently at the whole organism level to affect the production and function of all 3 organelles. Further, the increased lysosomal enzyme levels in lung of double mutant mice suggest a cause of a major clinical problem of HPS, lung fibrosis. Finally, doubly mutant HPS mice are a useful laboratory model for analysis of severe HPS phenotypes. PMID- 11861281 TI - Enhancement of the antitumor properties of interleukin-2 by its targeted delivery to the tumor blood vessel extracellular matrix. AB - Angiogenic processes depend on the precise coordination of different cell types and a complex exchange of signals, many of which derive from new specific components of the provisional, angiogenesis-related, extracellular matrix (ECM). Angiogenesis-associated ECM components thus represent appealing targets for the selective delivery of therapeutic molecules to newly forming tumor vessels. Results of a previous study indicated that a high affinity recombinant antibody (L19) to ED-B, a domain contained in the angiogenesis-associated isoform of fibronectin (B-FN), selectively and efficiently targets tumor vessels. The present study shows that a fusion protein between L19 and interleukin 2 (L19-IL 2) mediates the selective delivery and concentration of IL-2 to tumor vasculature, thereby leading to a dramatic enhancement of the therapeutic properties of the cytokine. By contrast, IL-2 fused to an irrelevant recombinant antibody used as a control fusion protein showed neither accumulation in tumors nor therapeutic efficacy. Tumors in mice treated with L19-IL-2 were significantly smaller compared to those in animals treated with saline, the control fusion protein, or IL-2 alone (P =.003,.003, and.002, respectively). Moreover, no significant differences in size were observed among the tumors from the different control groups (using the control fusion protein, a mixture of IL-2 and L19, or saline alone). Immunohistochemical analysis of tumor infiltrates demonstrated a significantly higher number of T lymphocytes, natural killer cells, and macrophages, as well as increased interferon-gamma (IFN-gamma) accumulation, in tumors from animals treated with L19-IL-2 compared to tumors from control groups. The fact that ED-B is 100% homologous in human and mouse, thus ensuring that L19 reacts equally well with human and murine antigen, should ultimately expedite transfer of this reagent to clinical trials. PMID- 11861282 TI - Increased sensitivity of T lymphocytes to tumor necrosis factor receptor 1 (TNFR1)- and TNFR2-mediated apoptosis in HIV infection: relation to expression of Bcl-2 and active caspase-8 and caspase-3. AB - The destruction of CD4 T cells in human immunodeficiency virus (HIV) infection is associated with activation of apoptotic programs, partly mediated by death receptors. The role of CD95L/CD95 in depletion of patients' CD4 T cells is well documented, but the possible contribution of the tumor necrosis factor/tumor necrosis factor receptor (TNF/TNFR) pathway has not been examined. In this study, we found that both TNFR1 and TNFR2 induced marked apoptosis in peripheral T cells from HIV-infected persons, involving both CD4 and CD8 T cells. Longitudinal follow-up of HIV(+) patients suggests an association between the in vivo evolution of CD4 T-cell numbers and variations in susceptibility to TNFR-induced apoptosis. Analysis of molecular mechanisms involved showed that it was not related to altered ex vivo expression of TNFR1-associated death domain, receptor interacting protein, or TNFR-associated factor 2. Susceptibility to TNFR-mediated apoptosis was rather related to Bcl-2 expression, because patients' T cells expressing high levels of Bcl-2 were completely protected from TNFR1- and TNFR2 induced cell death, whereas T cells expressing normal levels of Bcl-2 were not protected in patients in contrast to controls. Early recruitment of caspase-8 and caspase-3 is needed to transduce the apoptotic signals, and expression of both caspases in their active form was detected in blood T cells from HIV(+) patients, whereas it was hardly detected in controls. Moreover, ligation of TNFRs induced increased activation of both caspases in patients' T cells. Together these data demonstrate that exacerbated TNFR-mediated cell death of T cells from HIV infected individuals is associated with both alteration of Bcl-2 expression and activation of caspase-8 and caspase-3 and may contribute to the pathogenesis of acquired immunodeficiency syndrome. PMID- 11861283 TI - In vivo manipulation of dendritic cells to induce therapeutic immunity. AB - Efficient antigen presentation and T-cell priming are essential components of effective antitumor immunity. Dendritic cells are critical to both of these functions but to date no method has been devised that both targets antigen to these cells and activates them, in situ, in a manner that induces systemic immunity. In this study we combined a dendritic cell growth factor, Flt3 ligand, with a dendritic cell activator, immunostimulatory DNA, and a tumor antigen to activate and load dendritic cells in vivo. Initial studies showed that immunostimulatory DNA not only activates dendritic cells but also prolongs their survival in vivo and in vitro. Following treatment of mice with Flt3 ligand, coadministration of immunostimulatory DNA and antigen induced potent antitumor immunity, resulting in both tumor prevention and regression of existing tumors. CD8 cytotoxic T lymphocytes but not CD4 T cells were required for tumor protection. Natural killer cells also contributed to tumor protection. These results show that dendritic cells can be loaded with antigen and activated, in situ, and provide the basis for dendritic cell- targeted clinical strategies. PMID- 11861284 TI - Cell surface antigen CD109 is a novel member of the alpha(2) macroglobulin/C3, C4, C5 family of thioester-containing proteins. AB - Cell surface antigen CD109 is a glycosylphosphatidylinositol (GPI)-linked glycoprotein of approximately 170 kd found on a subset of hematopoietic stem and progenitor cells and on activated platelets and T cells. Although it has been suggested that T-cell CD109 may play a role in antibody-inducing T-helper function and it is known that platelet CD109 carries the Gov alloantigen system, the role of CD109 in hematopoietic cells remains largely unknown. As a first step toward elucidating the function of CD109, we have isolated and characterized a human CD109 cDNA from KG1a and endothelial cells. The isolated cDNA comprises a 4335 bp open-reading frame encoding a 1445 amino acid (aa) protein of approximately 162 kd that contains a 21 aa N-terminal leader peptide, 17 potential N-linked glycosylation sites, and a C-terminal GPI anchor cleavage addition site. We report that CD109 is a novel member of the alpha 2 macroglobulin (alpha 2M)/C3, C4, C5 family of thioester-containing proteins, and we demonstrate that native CD109 does indeed contain an intact thioester. Analysis of the CD109 aa sequence suggests that CD109 is likely activated by proteolytic cleavage and thereby becomes capable of thioester-mediated covalent binding to adjacent molecules or cells. In addition, the predicted chemical reactivity of the activated CD109 thioester is complement-like rather than resembling that of alpha 2M proteins. Thus, not only is CD109 potentially capable of covalent binding to carbohydrate and protein targets, but the t(1/2) of its activated thioester is likely extremely short, indicating that CD109 action is highly restricted spatially to the site of its activation. PMID- 11861285 TI - A tyrosine703serine polymorphism of CD109 defines the Gov platelet alloantigens. AB - The biallelic platelet-specific Gov antigen system-implicated in refractoriness to platelet transfusion, neonatal alloimmune thrombocytopenia, and posttransfusion purpura-is carried by the glycosylphosphatidylinositol (GPI) linked protein CD109. The recent identification of the human CD109 complementary DNA (cDNA) has allowed the molecular nature of the Gov alleles to be elucidated. By using reverse transcriptase-polymerase chain reaction (RT-PCR) to amplify CD109 cDNAs from 6 phenotypically homozygous Gov(aa) and Gov(bb) individuals, we have determined that the Gov alleles differ by an A to C single nucleotide polymorphism (SNP) at position 2108 of the coding region, resulting in a Tyr/Ser substitution at CD109 amino acid 703. Allele-specific PCR sequence-specific primers (SSP), PCR-restriction fragment length polymorphism, and real-time PCR studies of 15 additional donors (5 Gov(aa), 5 Gov(bb), and 5 Gov(ab)) confirmed that this SNP correlates with the Gov phenotype. In addition, Chinese hamster ovary cells transiently expressing nucleotide 2108 A>C CD109 cDNA variants were recognized specifically by allele-specific Gov antisera, indicating that this polymorphism defines the Gov alloantigenic determinants. Real-time PCR was then used to genotype 85 additional Gov phenotyped donors. In all but 3 cases, genomic testing concurred with the Gov phenotype. Repeat testing corrected 2 of these discrepancies in favor of the genotyping result. The third discrepancy could not be resolved, likely reflecting low-level CD109 expression below the sensitivity of the phenotyping assay. We conclude that the Gov alleles are defined by a 2108 A>C SNP that results in a Tyr703Ser substitution of CD109 and that genotyping studies are more accurate for Gov alloantigen determination than are conventional serologic methods. PMID- 11861286 TI - Requirement for MD-1 in cell surface expression of RP105/CD180 and B-cell responsiveness to lipopolysaccharide. AB - RP105 is a B-cell surface molecule that has been recently assigned as CD180. RP105 ligation with an antibody induces B-cell activation in humans and mice, leading to proliferation and up-regulation of a costimulatory molecule, B7.2/CD86. RP105 is associated with an extracellular molecule, MD-1. RP105/MD-1 has structural similarity to Toll-like receptor 4 (TLR4)/MD-2. TLR4 signals a membrane constituent of Gram-negative bacteria, lipopolysaccharide (LPS). MD-2 is indispensable for TLR4-dependent LPS responses because cells expressing TLR4/MD 2, but not TLR4 alone, respond to LPS. RP105 also has a role in LPS responses because B cells lacking RP105 show hyporesponsiveness to LPS. Little is known, however, regarding whether MD-1 is important for RP105-dependent LPS responses, as MD-2 is for TLR4. To address the issue, we developed mice lacking MD-1 and generated monoclonal antibodies (mAbs) to the protein. MD-1-null mice showed impairment in LPS-induced B-cell proliferation, antibody production, and B7.2/CD86 up-regulation. These phenotypes are similar to those of RP105-null mice. The similarity was attributed to the absence of cell surface RP105 on MD-1 null B cells. MD-1 is indispensable for cell surface expression of RP105. A role for MD-1 in LPS responses was further studied with anti-mouse MD-1 mAbs. In contrast to highly mitogenic anti-RP105 mAbs, the mAbs to MD-1 were not mitogenic but antagonistic on LPS-induced B-cell proliferation and on B7.2 up-regulation. Collectively, MD-1 is important for RP105 with respect to B-cell surface expression and LPS recognition and signaling. PMID- 11861287 TI - Soluble HLA class I molecules induce natural killer cell apoptosis through the engagement of CD8: evidence for a negative regulation exerted by members of the inhibitory receptor superfamily. AB - Herein, we show that CD8(dull), CD8(intermediate), and CD8(bright) natural killer (NK) cell clones can be identified. Triggering of CD8 with its natural ligand(s), represented by soluble HLA class I (sHLA-I), isolated either from serum of healthy donors or from HLA-I(-) 721.221 lymphoblastoid cell line transfected with HLA-A2, -Cw4, and -Bw46 alleles, or HLA-G1 leads to NK cell apoptosis. The magnitude of this effect directly correlated with the level of CD8 expression. sHLA-I-induced apoptosis depends on the interaction with CD8, as it was inhibited by masking this molecule with specific monoclonal antibodies (mAbs). Moreover, sHLA-I or CD8 cross-linking with specific mAbs elicited intracellular calcium increases, Fas ligand (FasL) messenger RNA transcription, and FasL secretion, which were needed for delivering the death signal. Indeed, this apoptosis was inhibited by preincubation of NK cell clones with Fas or FasL antagonist mAbs, indicating that the Fas/FasL pathway is involved. Furthermore, members of the inhibitory receptor superfamily, such as CD94/NKG2 complex or killer inhibitory receptors, were shown to exert an inhibitory effect on sHLA-I-mediated apoptosis and secretion of FasL. These findings suggest that interaction between sHLA-I and CD8 evokes an apoptotic signal that is down-regulated by inhibitory receptor superfamily that function as survival receptors in NK cells. PMID- 11861288 TI - Dendritic cells exposed to extracellular adenosine triphosphate acquire the migratory properties of mature cells and show a reduced capacity to attract type 1 T lymphocytes. AB - We previously reported that chronic stimulation with low, noncytotoxic doses of extracellular adenosine triphosphate (ATP) induced a distorted maturation of dendritic cells (DCs) and impaired their capacity to initiate T-helper (Th) 1 responses in vitro. Here, we examined the effects of ATP on chemokine-receptor expression and chemokine production by DCs. ATP strongly induced expression of CXC chemokine receptor 4 on both immature and lipopolysaccharide (LPS)-stimulated DCs and slightly up-regulated CC chemokine receptor (CCR) 7 on both DC types. In contrast, ATP reduced CCR5 expression on immature DCs. These effects were confirmed at both the messenger RNA and protein levels and were not produced by uridine triphosphate (UTP). Consistent with the changed receptor expression, ATP increased migration and intracellular calcium of immature and mature DCs to stromal-derived factor 1 (CXC ligand [CXCL] 12) and macrophage inflammatory protein [MIP] 3 beta (CC ligand [CCL] 19), whereas responses to MIP-1 beta (CCL4) were reduced. DCs are an important source of chemokines influencing recruitment of distinct T-lymphocyte subsets. ATP, but not UTP, significantly reduced LPS induced production of interferon-inducible protein 10 (CXCL10) and regulated upon activation, normal T-cell expressed and secreted chemokine (CCL5); increased secretion of macrophage-derived chemokine (CCL22); and did not change production of thymus and activation-regulated chemokine (CCL17). Consistent with these findings, supernatants from ATP-treated mature DCs attracted Th1 and T-cytotoxic 1 cells less efficiently, whereas migration of Th2 and T cytotoxic 2 cells was not affected. Our data suggest that ATP provides a signal for enhanced lymph node localization of DCs but that it may, at the same time, diminish the capacity of DCs to amplify type 1 immune responses. PMID- 11861289 TI - Analysis of natural killer cells in TAP2-deficient patients: expression of functional triggering receptors and evidence for the existence of inhibitory receptor(s) that prevent lysis of normal autologous cells. AB - Natural killer (NK) cells are characterized by the ability to kill cells that lack HLA class I molecules while sparing autologous normal (HLA class I(+)) cells. However, patients with transporter-associated antigen processing (TAP) deficiency, though displaying strong reductions of HLA class I surface expression, in most instances do not experience NK-mediated autoimmune phenomena. A possible mechanism by which TAP(-/-) NK cells avoid autoreactivity against autologous HLA class I-deficient cells could be based on either quantitative or qualitative defects of surface receptors involved in NK cell triggering. In this study we show that NK cells derived from 2 patients with TAP2(-/-) express normal levels of all known triggering receptors. As revealed by the analysis of polyclonal and clonal NK cells, these receptors display normal functional capabilities and allow the killing of a panel of NK-susceptible targets, including autologous B-LCLs. On the other hand, TAP2(-/-) NK cells were unable to kill either allogeneic (HLA class I(+)) or autologous (HLA class I(-) ) phytohemagglutinin (PHA) blasts even in the presence of anti-HLA class I monoclonal antibody. These data suggest that TAP2(-/-) NK cells express still unknown inhibitory receptor(s) capable of down-regulating the NK cell cytotoxicity on binding to surface ligand(s) expressed by T cell blasts. Functional analyses, both at the polyclonal and at the clonal level, are consistent with the concept that the putative inhibitory receptor is expressed by virtually all TAP2(-/-) NK cells, whereas it is present only in rare NK cells from healthy persons. Another possibility would be that TAP2(-/-) NK cells are missing a still unidentified triggering receptor involved in NK cell-mediated killing of PHA blasts. PMID- 11861290 TI - Quantitation, selection, and functional characterization of Epstein-Barr virus specific and alloreactive T cells detected by intracellular interferon-gamma production and growth of cytotoxic precursors. AB - Techniques for the quantitation of virus-specific and alloantigen-reactive T cells vary in their measurement of clinically relevant T-cell effector populations, their sensitivity and quantitative accuracy, and the time required to obtain measurable results. We compared frequencies of Epstein-Barr virus (EBV) specific and major alloantigen-reactive T cells as measured by flow cytometric analysis of responding T cells producing intracellular interferon-gamma (IFN gamma) and by limiting-dilution analysis (LDA) of cytotoxic T-cell precursors (CTLp) at sequential time points during the generation of EBV-specific T-cell lines. The expansion of EBV-specific T lymphocytes and the depletion of alloreactive T cells in cultures of T cells sensitized with autologous EBV transformed targets followed similar kinetics when measured by either method. Frequencies of EBV- specific T cells generating intracellular IFN-gamma exceeded by 25- to 90-fold the frequencies of responding CTLp at each stage of expansion, whereas the frequencies of alloreactive T cells generating intracellular IFN gamma exceeded by 30- to 220-fold those detected by LDA. The assay that quantitated T cells producing IFN-gamma yielded more reproducible and precise results than LDA. Furthermore, frequencies detected by the enumeration of T cells responding to immunodominant EBNA 3a and EBNA 3c peptides by IFN-gamma production or their capacity to bind peptide-HLA tetramers were strikingly similar and represented significant fractions of T cells generating IFN-gamma in response to autologous EBV B lymphoblastoid cell line. Functional analysis of responding viable T cells, fractionated on the basis of their secretion of IFN-gamma, demonstrated that EBV-specific and alloantigen cytotoxic T cells were predominantly or exclusively detected in the CD8(+)IFN-gamma(+) fraction of T cells. Strikingly, the CD4(+)IFN-gamma(+) cell fractions were not cytotoxic against EBV-transformed or allogeneic targets. PMID- 11861291 TI - The c-KIT mutation causing human mastocytosis is resistant to STI571 and other KIT kinase inhibitors; kinases with enzymatic site mutations show different inhibitor sensitivity profiles than wild-type kinases and those with regulatory type mutations. AB - Mutations of c-KIT causing spontaneous activation of the KIT receptor kinase are associated with sporadic adult human mastocytosis (SAHM) and with human gastrointestinal stromal tumors. We have classified KIT-activating mutations as either "enzymatic site" type (EST) mutations, affecting the structure of the catalytic portion of the kinase, or as "regulatory" type (RT) mutations, affecting regulation of an otherwise normal catalytic site. Using COS cells expressing wild-type or mutant KIT, 2 compounds, STI571 and SU9529, inhibited wild-type and RT mutant KIT at 0.1 to 1 microM but did not significantly inhibit the Asp816Val EST mutant associated with SAHM, even at 10 microM. Using 2 subclones of the HMC1 mast cell line, which both express KIT with an identical RT mutation but which differ in that one also expresses the Asp816Val EST mutation, both compounds inhibited the RT mutant KIT, thereby suppressing proliferation and producing apoptosis in the RT mutant-only cell line. Neither compound suppressed activation of Asp816Val EST mutant KIT, and neither produced apoptosis or significantly suppressed proliferation of the cell line expressing the Asp816Val mutation. These studies suggest that currently available KIT inhibitors may be useful in treating neoplastic cells expressing KIT activated by its natural ligand or by RT activating mutations such as gastrointestinal stromal tumors but that neither compound is likely to be effective against SAHM. Furthermore, these results help establish a general paradigm whereby classification of mutations affecting oncogenic enzymes as RT or EST may be useful in predicting tumor sensitivity or resistance to inhibitory drugs. PMID- 11861292 TI - Global gene expression profiling of multiple myeloma, monoclonal gammopathy of undetermined significance, and normal bone marrow plasma cells. AB - Bone marrow plasma cells (PCs) from 74 patients with newly diagnosed multiple myeloma (MM), 5 with monoclonal gammopathy of undetermined significance (MGUS), and 31 healthy volunteers (normal PCs) were purified by CD138(+) selection. Gene expression of purified PCs and 7 MM cell lines were profiled using high-density oligonucleotide microarrays interrogating about 6800 genes. On hierarchical clustering analysis, normal and MM PCs were differentiated and 4 distinct subgroups of MM (MM1, MM2, MM3, and MM4) were identified. The expression pattern of MM1 was similar to normal PCs and MGUS, whereas MM4 was similar to MM cell lines. Clinical parameters linked to poor prognosis, abnormal karyotype (P =.002) and high serum beta(2)-microglobulin levels (P =.0005), were most prevalent in MM4. Also, genes involved in DNA metabolism and cell cycle control were overexpressed in a comparison of MM1 and MM4. In addition, using chi(2) and Wilcoxon rank sum tests, 120 novel candidate disease genes were identified that discriminate normal and malignant PCs (P <.0001); many are involved in adhesion, apoptosis, cell cycle, drug resistance, growth arrest, oncogenesis, signaling, and transcription. A total of 156 genes, including FGFR3 and CCND1, exhibited highly elevated ("spiked") expression in at least 4 of the 74 MM cases (range, 4 25 spikes). Elevated expression of these 2 genes was caused by the translocation t(4;14)(p16;q32) or t(11;14)(q13;q32). Thus, novel candidate MM disease genes have been identified using gene expression profiling and this profiling has led to the development of a gene-based classification system for MM. PMID- 11861293 TI - TEL/platelet-derived growth factor receptor beta activates phosphatidylinositol 3 (PI3) kinase and requires PI3 kinase to regulate the cell cycle. AB - TEL/platelet-derived growth factor receptor beta (PDGF beta R) is the protein product of the t(5;12) translocation in chronic myelomonocytic leukemia. TEL/PDGF beta R transforms interleukin-3 (IL-3)-dependent Ba/F3 and 32D cells to IL-3 independence and induces a murine myeloproliferative disease in a bone marrow transplantation model of leukemogenesis. The fusion protein encodes a constitutively activated, cytoplasmic tyrosine kinase that activates multiple signal transduction pathways. To identify the signaling pathways that are necessary for transformation by TEL/PDGF beta R, transformed Ba/F3 and 32D cells were studied. TEL/PDGF beta R activates the kinase activity of phosphatidylinositol-3 (PI3) kinase and stimulates phosphorylation of its downstream substrates, including Akt and p70S6 kinase. Activation of this pathway requires the kinase activity of TEL/PDGF beta R and is inhibited by the PDGF beta R inhibitor, STI571. Furthermore, inhibition of PI3 kinase with the pharmacologic inhibitor, LY294002, inhibits growth of the transformed cells. Treated cells arrest in the G1 phase of the cell cycle within 16 hours but do not undergo apoptosis. To study the mechanism of cell cycle arrest by LY294002, the activity of the cdk4 complex, which regulates the transit of cells from the G1 to S phase in hematopoietic cells, was examined. Both STI571 and LY294002 lead to a decrease in the activity of cdk4 kinase activity and a decrease in expression of both Cyclin D2 and Cyclin E within several hours. These studies demonstrate the presence of a signaling pathway from TEL/PDGF beta R to PI3 kinase and subsequently to regulation of the cdk4 kinase complex. Activation of this pathway is necessary for transformation by TEL/PDGF beta R. PMID- 11861294 TI - Overexpression of SOCS-2 in advanced stages of chronic myeloid leukemia: possible inadequacy of a negative feedback mechanism. AB - Constitutive activation of the BCR-ABL tyrosine kinase is fundamental to the pathogenesis of chronic myeloid leukemia (CML). STI571 inhibits this activity and modulates the transcription of several genes. It was shown by differential display that the suppressor of cytokine signaling-2 (SOCS-2) gene was down regulated by STI571 treatment in 14 of 16 BCR-ABL-positive cell lines and in 2 BCR-ABL-transfected murine lines, but not in BCR-ABL-negative counterparts. The effect was maximal at 2 hours and persisted for at least 24 hours after exposure to 1 microM STI571, whereas SOCS-1 and SOCS-3 expression were unaffected. Baseline levels of SOCS-2 were significantly higher in BCR-ABL-positive as compared with BCR-ABL-negative cell lines. It was similar in leukocytes and CD34(+) cells from healthy persons (n = 44) and patients with CML in chronic phase (CP; n = 60) but significantly increased in patients with CML in blast crisis (BC; n = 20) (P <.0001). Mononuclear cells (MNCs) from 3 of 4 patients with CML in BC showed a 2-fold to 12-fold down-regulation of SOCS-2 levels on in vitro exposure to STI571; moreover, a 2-fold to 11-fold decrease in SOCS-2 was observed in MNCs from 7 of 8 patients with CML in BC who responded to treatment with STI571. Refractoriness to STI571 or relapse after initial response was accompanied by augmentation of SOCS-2 expression. Ectopic overexpression of SOCS 2 in 32Dp210 cells slowed growth, inhibited clonogenicity, and increased their motility and sensitivity to STI571. Overall, the results suggest that SOCS-2 is a component of a negative feedback mechanism; it is induced by Bcr-Abl but cannot reverse its overall growth-promoting effects in blastic transformation. PMID- 11861296 TI - Secretion of heparin-binding protein from human neutrophils is determined by its localization in azurophilic granules and secretory vesicles. AB - Human neutrophils have an important role in host defense against microbial infection. At different stages of an infectious process, neutrophils progressively up-regulate receptors and release various effector molecules. These are stored in several distinct types of granules with varying propensity to be secreted. Heparin-binding protein (HBP), also known as CAP37 or azurocidin, is a multifunctional, inactive serine-protease homologue. The present work shows that HBP is released from neutrophils on stimulation with secretagogues that do not trigger the secretion of azurophilic granule content. Therefore, the subcellular localization of HBP was investigated in more detail. Immunofluorescence microscopy revealed that HBP was localized close to the plasma membrane. Further analysis by fractionation of postnuclear supernatants from cavitated neutrophils showed that HBP is stored in azurophilic granules and secretory vesicles but that it is also detected to a minor extent in the plasma membrane. These findings were confirmed by immunoelectron microscopy showing that HBP colocalized with marker proteins of azurophilic granules and secretory vesicles. The presence of HBP in secretory vesicles possibly depends on the stage of cell differentiation, since the promyelocytic cell line HL-60 contains less HBP than mature neutrophils, stored exclusively in the less easily mobilized azurophilic granules. Our findings suggest that HBP can be synthesized or targeted to easily mobilized compartments at a late stage of neutrophil maturation. The ability of neutrophils to secrete HBP from secretory vesicles may be important for proinflammatory functions of this protein, such as the alteration of vascular permeability. PMID- 11861295 TI - Evidence for position effects as a variant ETV6-mediated leukemogenic mechanism in myeloid leukemias with a t(4;12)(q11-q12;p13) or t(5;12)(q31;p13). AB - The ETV6 gene (first identified as TEL) is a frequent target of chromosomal translocations in both myeloid and lymphoid leukemias. At present, more than 40 distinct translocations have been cytogenetically described, of which 13 have now also been characterized at the molecular level. These studies revealed the generation of in-frame fusion genes between different domains of ETV6 and partner genes encoding either kinases or transcription factors. However, in a number of cases-including a t(6;12)(q23;p13), the recurrent t(5;12)(q31;p13), and some cases of the t(4;12)(q11-q12;p13) described in this work-functionally significant fusions could not be identified, raising the question as to what leukemogenic mechanism is implicated in these cases. To investigate this, we have evaluated the genomic regions at 4q11-q12 and 5q31, telomeric to the breakpoints of the t(4;12)(q11-q12;p13) and t(5;12)(q31;p13). The homeobox gene GSH2 at 4q11-q12 and the IL-3/CSF2 locus at 5q31 were found to be located close to the respective breakpoints. In addition, GSH2 and IL-3 were found to be ectopically expressed in the leukemic cells, suggesting that expression of GSH2 and IL-3 was deregulated by the translocation. Our results indicate that, besides the generation of fusion transcripts, deregulation of the expression of oncogenes could be a variant leukemogenic mechanism for translocations involving the 5' end of ETV6, especially for those translocations lacking functionally significant fusion transcripts. PMID- 11861297 TI - Macrophage functional maturation and cytokine production are impaired in C/EBP epsilon-deficient mice. AB - Members of the CCAAT/enhancer-binding protein (C/EBP) family are involved in the regulation of cellular differentiation and function of many tissues. Unlike the other members of the family, C/EBP epsilon expression is restricted to granulocytes, macrophages, and lymphocytes. C/EBP epsilon is highly conserved between human and rodents and is essential for terminal granulopoiesis in both species. To study the role that C/EBP epsilon plays in macrophages, wild-type and C/EBP epsilon-deficient (-/-) murine macrophages obtained from thioglycollate elicited peritoneal lavages and differentiated bone marrow cells were compared. Although macrophage development occurred in both types of mice, the C/EBP epsilon -/- cells had a lower expression of macrophage markers and a morphologic and ultrastructural appearance of immaturity. Phagocytic function, measured by calculating the percentage of internalized opsonized fluorescein isothiocyanate (FITC)-labeled yeast, was significantly impaired in the C/EBP epsilon -/- macrophages compared with their wild-type counterparts. Furthermore, the differential expression of 26 macrophage-specific genes between wild-type and C/EBP-/- mice was analyzed. A subset of genes involved in differentiation, immune, and inflammatory responses was found down-regulated in the C/EBP-/- macrophages. Taken together, this study implicates the C/EBP epsilon gene as an important transcription factor required for normal function and development of macrophages. PMID- 11861298 TI - Defects in leukocyte-mediated initiation of lipid peroxidation in plasma as studied in myeloperoxidase-deficient subjects: systematic identification of multiple endogenous diffusible substrates for myeloperoxidase in plasma. AB - More than a decade ago it was demonstrated that neutrophil activation in plasma results in the time-dependent formation of lipid hydroperoxides through an unknown, ascorbate-sensitive pathway. It is now shown that the mechanism involves myeloperoxidase (MPO)-dependent use of multiple low-molecular-weight substrates in plasma, generating diffusible oxidant species. Addition of activated human neutrophils (from healthy subjects) to plasma (50%, vol/vol) resulted in the peroxidation of endogenous plasma lipids by catalase-, heme poison-, and ascorbate-sensitive pathways, as assessed by high-performance liquid chromatography (HPLC) with on-line electrospray ionization tandem mass spectrometric analysis of free and lipid-bound 9-HETE and 9-HODE. In marked contrast, neutrophils isolated from multiple subjects with MPO deficiency failed to initiate peroxidation of plasma lipids, but they did so after supplementation with isolated human MPO. MPO-dependent use of a low-molecular-weight substrate(s) in plasma for initiating lipid peroxidation was illustrated by demonstrating that the filtrate of plasma (10-kd MWt cutoff) could supply components required for low-density lipoprotein lipid peroxidation in the presence of MPO and H(2)O(2). Subsequent HPLC fractionation of plasma filtrate (10-kd MWt cutoff) by sequential column chromatography identified nitrite, tyrosine, and thiocyanate as major endogenous substrates and 17 beta-estradiol as a novel minor endogenous substrate in plasma for MPO in promoting peroxidation of plasma lipids. These results strongly suggest that the MPO-H(2)O(2) system of human leukocytes serves as a physiological mechanism for initiating lipid peroxidation in vivo. PMID- 11861299 TI - Abnormally spliced beta-globin mRNAs: a single point mutation generates transcripts sensitive and insensitive to nonsense-mediated mRNA decay. AB - Nonsense-mediated mRNA decay (NMD) represents a phylogenetically widely conserved splicing- and translation-dependent mechanism that eliminates transcripts with premature translation stop codons and suppresses the accumulation of C-terminally truncated peptides. Elimination of frameshifted transcripts that result from faulty splicing may be an important function of NMD. To test this hypothesis directly, this study used the IVS1 + 5 G>A thalassemia mutation of the human beta globin gene as a model system. We generated beta-globin gene constructs with this mutation and an iron-responsive element in the 5' untranslated region, which allowed specific experimental activation and inactivation of translation and, hence, NMD of this transcript. Premessenger RNAs with IVS1 + 5 G>A were spliced at normal sites and cryptic sites, enabling a direct comparison of the effect of NMD on the accumulation of normal and frameshifted messenger RNAs. In transfected HeLa cells, the predominant frameshifted transcript was degraded under conditions of active NMD, whereas accumulation to high levels occurred under conditions of specifically disabled NMD, thereby indicating an important physiologic function of NMD in the control of the splicing process. An unexpected finding was that accumulation of a second aberrant transcript remained unaffected by NMD. The IVS1 + 5 G>A mutation thus revealed the presence of an unknown cis-acting determinant that influences the NMD sensitivity of a putative NMD substrate. It can therefore serve as a useful tool for defining the mechanisms that permit specific transcripts to circumvent the NMD pathway. PMID- 11861300 TI - Failure of red blood cell maturation in mice with defects in the high-density lipoprotein receptor SR-BI. AB - Mammalian erythrocytes undergo a unique maturation process in which they discard their nuclei and organelles and assume a flexible biconcave shape. We found that altered plasma lipoprotein metabolism can profoundly influence these events. Abnormal erythrocyte morphology was observed in hypercholesterolemic mice lacking the high-density lipoprotein receptor SR-BI. This was exacerbated by feeding mice a high-cholesterol diet or, more dramatically, by inactivating the apolipoprotein E gene. Erythrocytes from SR-BI(-/-)/apolipoprotein E(-/-) mice and SR-BI(-/-) mice that were fed cholesterol had markedly increased membrane cholesterol. Their morphology appeared immature, with macrocytosis, irregular shape, and large autophagolysosomes. Autophagolysosomes from SR-BI(-/-)/apolipoprotein E(-/-) erythrocytes were expelled when the erythrocytes were transfused into wild-type animals or incubated in vitro with normolipidemic serum or the cholesterol sequestering agent methyl cyclodextrin. We propose that autophagocytosis and phagolysosome expulsion are essential steps in erythroid maturation and that expulsion is inhibited in the presence of markedly increased cellular cholesterol. PMID- 11861301 TI - Flt3 ligand therapy for recipients of allogeneic bone marrow transplants expands host CD8 alpha(+) dendritic cells and reduces experimental acute graft-versus host disease. AB - Recent evidence suggests that dendritic cells (DCs) can regulate and amplify immune responses. Flt3 ligand (FL)-derived DC function was tested as a stimulator of allogeneic lymphocytes in vitro and in vivo. Treatment of mice with FL dramatically expanded DC number, but DCs isolated from FL-treated mice (FL DCs) were poor stimulators of allogeneic T-cell responses in vitro. Further activation of FL DCs did not restore their stimulatory ability, and FL DCs did not suppress the stimulation of the allogeneic T cells by normal DCs. FL treatment significantly increased the CD8 alpha(+) DC subset, which appeared to be the reason for their poor stimulatory capacity. These observations were confirmed in vivo using a mouse model of acute graft-versus-host disease (GVHD) wherein host DCs play a critical role. FL treatment of recipients before allogeneic bone marrow transplantation dramatically suppressed donor T-cell responses to host antigens, thereby reducing GVHD mortality (P <.01). These data represent a novel strategy that alters host DCs and reduces acute GVHD. PMID- 11861302 TI - HPA-1a phenotype-genotype discrepancy reveals a naturally occurring Arg93Gln substitution in the platelet beta 3 integrin that disrupts the HPA-1a epitope. AB - A single nucleotide polymorphism (SNP) at position 196 in the beta 3 integrin causes a Leu33Pro substitution in the mature protein. Alloimmunization against the beta 3Leu33 form (human platelet antigen [HPA]-1a, Pl(A1), Zw(a)) in patients who are beta 3Pro33 homozygous (HPA-1b1b, Pl(A2A2), Zw(bb)) causes neonatal alloimmune thrombocytopenia, posttransfusion purpura, or refractoriness to platelet transfusion. Studies with recombinant proteins have demonstrated that amino acids 1 to 66 and 288 to 490 of the beta 3 integrin contribute to HPA-1a epitope formation. In determining the HPA-1a status of more than 6000 donors, we identified a donor with an HPA-1a(weak) phenotype and an HPA-1a1b genotype. The platelets from this donor had normal levels of surface alpha IIb beta 3 but reacted only weakly with monoclonal and polyclonal anti-HPA-1a by whole blood enzyme-linked immunosorbent assay (ELISA), flow cytometry, and sandwich ELISA. We reasoned that an alteration in the primary nucleotide sequence of the beta 3Leu33 allele of this donor was disrupting the HPA-1a epitope. In agreement with this hypothesis, sequencing platelet RNA-derived alpha IIb and beta 3 cDNA identified a novel G/A SNP at position 376 of the beta 3 integrin that encodes for an Arg93Gln replacement in the beta 3Leu33 allele. Coexpression of the beta 3Leu33Gln93 encoding cDNA in Chinese hamster ovary cells with human alpha IIb cDNA showed that the surface-expressed alpha IIb beta 3 reacted normally with beta 3 integrin-specific monoclonal antibodies but only weakly with monoclonal anti-HPA-1a. Our results show that an Arg93Gln mutation in the beta 3Leu33 encoding allele disrupts the HPA-1a epitope, suggesting that Arg93 contributes to the formation of the HPA-1a B-cell epitope. PMID- 11861303 TI - A cure for murine sickle cell disease through stable mixed chimerism and tolerance induction after nonmyeloablative conditioning and major histocompatibility complex-mismatched bone marrow transplantation. AB - The morbidity and mortality associated with sickle cell disease (SCD) is caused by hemolytic anemia, vaso-occlusion, and progressive multiorgan damage. Bone marrow transplantation (BMT) is currently the only curative therapy; however, toxic myeloablative preconditioning and barriers to allotransplantation limit this therapy to children with major SCD complications and HLA-matched donors. In trials of myeloablative BMT designed to yield total marrow replacement with donor stem cells, a subset of patients developed mixed chimerism. Importantly, these patients showed resolution of SCD complications. This implies that less toxic preparative regimens, purposefully yielding mixed chimerism after transplantation, may be sufficient to cure SCD without the risks of myeloablation. To rigorously test this hypothesis, we used a murine model for SCD to investigate whether nonmyeloablative preconditioning coupled with tolerance induction could intentionally create mixed chimerism and a clinical cure. We applied a well-tolerated, nonirradiation-based, allogeneic transplantation protocol using nonmyeloablative preconditioning (low-dose busulfan) and costimulation blockade (CTLA4-Ig and anti-CD40L) to produce mixed chimerism and transplantation tolerance to fully major histocompatibility complex-mismatched donor marrow. Chimeric mice were phenotypically cured of SCD and had normal RBC morphology and hematologic indices (hemoglobin, hematocrit, reticulocyte, and white blood cell counts) without evidence of graft versus host disease. Importantly, they also showed normalization of characteristic spleen and kidney pathology. These experiments demonstrate the ability to produce a phenotypic cure for murine SCD using a nonmyeloablative protocol with fully histocompatibility complex-mismatched donors. They suggest a future treatment strategy for human SCD patients that reduces the toxicity of conventional BMT and expands the use of allotransplantation to non-HLA-matched donors. PMID- 11861304 TI - Calpain is a signal transducer and activator of transcription (STAT) 3 and STAT5 protease. AB - Truncation of signal transducer and activator of transcription (STAT) 5 at the carboxy-terminal domain, either by genetic engineering or by proteolytic cleavage, results in generation of dominant-negative forms. A nuclear serine protease expressed in the myeloid precursor cells is known to mediate this cleavage, but other proteases responsible for this reaction were unknown. We found that calpain, a ubiquitously expressed cysteine protease, also trims STAT5 in vivo and in vitro, within the carboxy-terminal domain. Nuclear element is not necessary for calpain-mediated STAT5 cleavage, since this process occurs in platelets. We also found that STAT3 is a substrate for calpain in vivo and in vitro, indicating that calpain-mediated cleavage is a common feature of STAT3 and STAT5. Thus, our study reveals a novel pathway for posttranslational modification of STAT3 and STAT5. PMID- 11861305 TI - Immunophenotypic evaluation of the plasma cell compartment in multiple myeloma: a tool for comparing the efficacy of different treatment strategies and predicting outcome. AB - Multiparametric immunophenotyping can be a sensitive method for analyzing the plasma cell (PC) compartment in patients with multiple myeloma because it discriminates between myelomatous and normal PCs. Using this approach, we compared the efficacy of high-dose chemotherapy followed by autologous stem cell transplantation (ASCT) with that of conventional chemotherapy. We found that ASCT provided a significantly greater reduction in the level of residual tumor PCs and with better recovery of normal PCs. This profile of coexistence of normal PCs and myelomatous PCs resembled that observed in monoclonal gammopathy of undetermined significance. We also found that treatment-induced changes in the PC compartment correlated with disease outcome. Thus, patients in whom at least 30% of gated PCs had a normal phenotype after treatment had a significantly longer progression free survival (60 +/- 6 months versus 34 +/- 12 months; P =.02). PMID- 11861306 TI - Retrovirally mediated correction of bone marrow-derived mesenchymal stem cells from patients with mucopolysaccharidosis type I. AB - We have investigated the utility of bone marrow-derived mesenchymal stem cells (MSCs) as targets for gene therapy of the autosomal recessive disorder mucopolysaccharidosis type IH (MPS-IH, Hurler syndrome). Cultures of MSCs were initially exposed to a green fluorescent protein-expressing retrovirus. Green fluorescent protein-positive cells maintained their proliferative and differentiation capacity. Next we used a vector encoding alpha-L-iduronidase (IDUA), the enzyme that is defective in MPS-IH. Following transduction, MPS-IH MSCs expressed high levels of IDUA and secreted supernormal levels of this enzyme into the extracellular medium. Exogenous IDUA expression led to a normalization of glycosaminoglycan storage in MPS-IH cells, as evidenced by a dramatic decrease in the amount of (35)SO(4) sequestered within the heparan sulfate and dermatan sulfate compartments of these cells. Finally, gene-modified MSCs were able to cross-correct the enzyme defect in untransduced MPS-IH fibroblasts via protein transfer. PMID- 11861308 TI - Autocrine production of epithelial cell-derived neutrophil attractant-78 induced by granulocyte colony-stimulating factor in neutrophils. AB - Whereas mobilization to inflammatory sites is an important function of neutrophils, it remains to be determined whether granulocyte colony-stimulating factor (G-CSF) stimulates the mobilization of neutrophils to the inflammatory sites. This study compared the expression of more than 9000 genes in neutrophils treated with and without G-CSF with the use of a DNA microarray system to determine the effects of G-CSF on the function of neutrophils. It was found that messenger RNA expression of epithelial cell-derived neutrophil attractant-78 (ENA 78), which has been reported to be a chemotactic factor for neutrophils, was induced by G-CSF in neutrophils. The study demonstrated that the supernatant of G CSF-treated neutrophils induced the chemotaxis of neutrophils and that anti-ENA 78 antibody and anti-CXCR-2 antibody inhibited the chemotaxis. These data suggest that G-CSF may enhance the mobilization of neutrophils and consequently augment the accumulation of neutrophils in the inflammatory sites through the secretion of ENA-78. PMID- 11861307 TI - Ph(+) acute lymphoblastic leukemia resistant to the tyrosine kinase inhibitor STI571 has a unique BCR-ABL gene mutation. AB - The tyrosine kinase inhibitor STI571 is a promising agent for the treatment of advanced Philadelphia chromosome positive (Ph(+)) acute lymphoblastic leukemia (ALL), but resistance develops rapidly in most patients after an initial response. To identify mechanisms of resistance to STI571, 30 complementary DNAs (including 9 matched samples) obtained from the bone marrow of individuals with Ph(+) ALL were analyzed by direct sequencing of a 714-base pair region of ABL encoding for the adenosine triphosphate (ATP)-binding site and the kinase activation loop. A single point mutation was found at nucleotide 1127 (GI6382056) resulting in Glu255Lys. This mutation occurred in 6 of 9 patients (67%) following their treatment with STI571 but not in the samples from patients before beginning treatment with STI571. Glu255Lys is within the motif important for forming the pocket of the ATP-binding site in ABL and it is highly conserved across species. In conclusion, Ph(+) ALL samples resistant to STI571 have a unique mutation Glu255Lys of BCR-ABL. PMID- 11861309 TI - Requirement of Gab2 for mast cell development and KitL/c-Kit signaling. AB - Mast cells are thought to participate in a variety of immune responses, such as parasite resistance and the allergic reaction. Mast cell development depends on stem cell factor (Kit ligand) and its receptor, c-Kit. Gab2 is an adaptor molecule containing a pleckstrin homology domain and potential binding sites for SH2 and SH3 domains. Gab2 is phosphorylated on tyrosine after stimulation with cytokines and growth factors, including KitL. Gab2-deficient mice were created to define the physiological requirement for Gab2 in KitL/c-Kit signaling and mast cell development. In Gab2-deficient mice, the number of mast cells was reduced markedly in the stomach and less severely in the skin. Bone marrow-derived mast cells (BMMCs) from the Gab2-deficient mice grew poorly in response to KitL. KitL induced ERK MAP kinase and Akt activation were impaired in Gab2-deficient BMMCs. These data indicate that Gab2 is required for mast cell development and KitL/c Kit signaling. PMID- 11861310 TI - Allogenic fetal liver cells have a distinct competitive engraftment advantage over adult bone marrow cells when infused into fetal as compared with adult severe combined immunodeficient recipients. AB - In utero transplantation (IUT) is becoming a viable option for the treatment of various immune and metabolic disorders diagnosed early in gestation. In this study, donor fetal liver cells had a 10-fold competitive engraftment advantage relative to adult bone marrow in allogeneic fetal severe combined immunodeficient (SCID) recipients compared with adult recipients. In contrast, adult bone marrow cells engrafted slightly better than fetal liver cells in allogeneic adult SCID transplant recipients. By using different ratios of fetal and adult cell mixtures, fetal liver cells repopulated 8.2 times better than adult bone marrow cells in fetal recipients, but only 0.8 times as well in adult recipients. Fetal SCID recipients were more permissive to an allogeneic donor graft than adult recipients. These data indicate that the recipient microenvironment may regulate the engraftment efficiency of a given stem cell source and suggest that the use of cord blood should be tested in clinical IUT. PMID- 11861311 TI - P2 receptors: new potential players in atherosclerosis. AB - Atherosclerosis is a focal inflammatory disease of the arterial wall. It starts with the formation of fatty streaks on the arterial wall that evolve to form a raised plaque made of smooth muscle cells (SMCs), and infiltrating leukocytes surrounding a necrotic core. The pathogenesis of the atherosclerotic lesion is incompletely understood, but it is clear that a dysfunction of the endothelium, recruitment and activation of inflammatory cells and SMC proliferation have a pivotal role. Over recent years receptors for extracellular nucleotides, the P2 receptors, have been recognized as fundamental modulators of leukocytes, platelets, SMCs and endothelial cells. P2 receptors mediate chemotaxis, cytokine secretion, NO generation, platelet aggregation and cell proliferation in response to accumulation of nucleotides into the extracellular milieu. Clinical trials have shown the benefit of antagonists of the ADP platelet receptor(s) in the prevention of vascular accidents in patients with atherosclerosis. Therefore, we anticipate that a deeper understanding of the involvement of P2 receptors in atheroma formation will open new avenues for drug design and therapeutic intervention. PMID- 11861312 TI - Role of bradykinin in preconditioning and protection of the ischaemic myocardium. PMID- 11861314 TI - Role of endothelin ET(A)- and ET(B)-receptors in haemodynamic compensation following haemorrhage in anaesthetized rats. AB - 1. This study examined the role of endothelin ET(A) and ET(B) receptors on haemodynamic compensation following haemorrhage (-17.5 ml kg(-1)) in thiobutabarbitone-anaesthetized rats. Rats were divided into four groups (n=6 each): time-control, haemorrhage-control, haemorrhage after treatment with FR 139317 (ET(A)-receptor antagonist), and haemorrhage after treatment with BQ-788 (ET(B)-receptor antagonist). 2. In the time-control rats, there were no significant changes in any haemodynamics for the duration of the experiments. Relative to the time-control rats, rats given haemorrhage had reduced mean arterial pressure (MAP), cardiac output (CO) and mean circulatory filling pressure (MCFP), but increased systemic vascular resistance (R(SV)). Venous resistance (R(V)) was slightly (but insignificantly) reduced by haemorrhage. MAP, however, gradually returned towards baseline (-17+/-4 and -3+/-2 mmHg at 10 and 60 min after haemorrhage, respectively) as a result of a further increase in R(SV). 3. Pre-treatment with FR 139317 (i.v. 1 mg kg(-1), followed by 1 mg kg(-1) h(-1)) accentuated haemorrhage-induced hypotension through abolition of the increase in R(SV). FR 139317 did not modify haemorrhage-induced changes in CO, MCFP and R(V). 4. Pre-treatment of BQ-788 (3 mg kg(-1)) did not affect MAP or MCFP following haemorrhage; however, CO was lower, and R(SV) as well as R(V) were higher relative to the readings in the haemorrhaged-control rats. 5. These results show that following compensated haemorrhage, ET maintains arterial resistance and blood pressure via the activation of ET(A) but not ET(B) receptors. PMID- 11861315 TI - trans-4-Amino-2-methylbut-2-enoic acid (2-MeTACA) and (+/-)-trans-2 aminomethylcyclopropanecarboxylic acid ((+/-)-TAMP) can differentiate rat rho3 from human rho1 and rho2 recombinant GABA(C) receptors. AB - 1. This study investigated the effects of a number of GABA analogues on rat rho3 GABA(C) receptors expressed in Xenopus oocytes using 2-electrode voltage clamp methods. 2. The potency order of agonists was muscimol (EC(50)=1.9 +/- 0.1 microM) (+)-trans-3-aminocyclopentanecarboxylic acids ((+)-TACP; EC(50)=2.7 +/- 0.9 microM) trans-4-aminocrotonic acid (TACA; EC(50)=3.8 +/-0.3 microM) GABA (EC(50)=4.0 +/- 0.3 microM) > thiomuscimol (EC(50)=24.8 +/- 2.6 microM) > (+/-) cis-2-aminomethylcyclopropane-carboxylic acid ((+/-)-CAMP; EC(50)=52.6 +/-8.7 microM) > cis-4-aminocrotonic acid (CACA; EC(50)=139.4 +/- 5.2 microM). 3. The potency order of antagonists was (+/-)-trans-2-aminomethylcyclopropanecarboxylic acid ((+/-)-TAMP; K(B)=4.8+/-1.8 microM) (1,2,5,6-tetrahydropyridin-4 yl)methylphosphinic acid (TPMPA; K(B)=4.8 +/-0.8 microM) > (piperidin-4 yl)methylphosphinic acid (P4MPA; K(B)=10.2+/-2.3 microM) 4,5,6,7 tetrahydroisoxazolo[5,4-c]pyridin-3-ol (THIP; K(B)=10.2+/-0.3 microM) imidazole-4 acetic acid (I4AA; K(B)=12.6+/-2.7 microM) > 3-aminopropylphosphonic acid (3-APA; K(B)=35.8+/-13.5 microM). 4. trans-4-Amino-2-methylbut-2-enoic acid (2-MeTACA; 300 microM) had no effect as an agonist or an antagonist indicating that the C2 methyl substituent is sterically interacting with the ligand-binding site of rat rho3 GABA(C) receptors. 5. 2-MeTACA affects rho1 and rho2 but not rho3 GABA(C) receptors. In contrast, (plus minus)-TAMP is a partial agonist at rho1 and rho2 GABA(C) receptors, while at rat rho3 GABA(C) receptors it is an antagonist. Thus, 2-MeTACA and (+/-)-TAMP could be important pharmacological tools because they may functionally differentiate between rho1, rho2 and rho3 GABA(C) receptors in vitro. PMID- 11861316 TI - Cyclooxygenase-1 derived prostaglandins are involved in the maintenance of renal function in rats with cirrhosis and ascites. AB - 1. The maintenance of renal function in decompensated cirrhosis is highly dependent on prostaglandins (PGs). Since PG synthesis is mediated by cyclooxygenase-1 and -2 (COX-1 and COX-2), the present study was designed to examine which COX isoform is involved in this phenomenon. 2. Renal COX-1 and COX 2 protein expression and distribution were analysed by Western blot and immunohistochemistry in nine rats with carbon tetrachloride-induced cirrhosis and ascites and 10 control animals. The effects of placebo and selective COX-1 (SC 560) and COX-2 (celecoxib) inhibitors on urine flow (V), urinary excretion of sodium (U(Na)V) and PGE(2) (U(PGE2)V), glomerular filtration rate (GFR), renal plasma flow (RPF), the diuretic and natriuretic responses to furosemide and renal water metabolism were assessed in 88 rats with cirrhosis and ascites. 3. COX-1 protein levels were found to be unchanged in kidneys from cirrhotic rats. In contrast, these animals showed enhanced renal COX-2 protein expression which was focally increased in the corticomedullary region. Although U(PGE2)V was equally reduced by SC-560 and celecoxib, only SC-560 produced a significant decrease in U(Na)V, GFR and RPF and a pronounced impairment in the diuretic and natriuretic responses to furosemide in rats with cirrhosis and ascites. Neither SC-560 nor celecoxib affected renal water metabolism in cirrhotic rats. 4. These results indicate that despite abundant renal COX-2 protein expression, the maintenance of renal function in cirrhotic rats is mainly dependent on COX-1-derived prostaglandins. PMID- 11861317 TI - Inhibition of the NMDA response by pregnenolone sulphate reveals subtype selective modulation of NMDA receptors by sulphated steroids. AB - 1. The neurosteroid pregnenolone sulphate (PS) potentiates N-methyl-D-aspartate (NMDA) receptor mediated responses in various neuronal preparations. The NR1 subunit can combine with NR2A, NR2B, NR2C, or NR2D subunits to form functional receptors. Differential NR2 subunit expression in brain and during development raises the question of how the NR2 subunit influences NMDA receptor modulation by neuroactive steroids. 2. We examined the effects of PS on the four diheteromeric NMDA receptor subtypes generated by co-expressing the NR1(100) subunit with each of the four NR2 subunits in Xenopus oocytes. Whereas PS potentiated NMDA-, glutamate-, and glycine-induced currents of NR1/NR2A and NR1/NR2B receptors, it was inhibitory at NR1/NR2C and NR1/NR2D receptors. 3. In contrast, pregnanolone sulphate (3alpha5betaS), a negative modulator of the NMDA receptor that acts at a distinct site from PS, inhibited all four subtypes, but was approximately 4 fold more potent at NR1/NR2C and NR1/NR2D than at NR1/NR2A and NR1/NR2B receptors. 4. These findings demonstrate that residues on the NR2 subunit are key determinants of modulation by PS and 3alpha5betaS. The modulatory effects of PS, but not 3alpha5betaS, on dose-response curves for NMDA, glutamate, and glycine are consistent with a two-state model in which PS either stabilizes or destabilizes the active state of the receptor, depending upon which NR2 subunit is present. 5. The selectivity of sulphated steroid modulators for NMDA receptors of specific subunit composition is consistent with a neuromodulatory role for endogenous sulphated steroids. The results indicate that it may be possible to develop therapeutic agents that target steroid modulatory sites of specific NMDA receptor subtypes. PMID- 11861318 TI - A flavonoid-rich diet increases nitric oxide production in rat aorta. AB - 1. Red wine intake is associated with a low risk of cardiovascular disease. This effect has been partly attributed to the action of polyphenolic compounds, which decrease the oxidation of plasma low density lipoproteins. Moreover, nitric oxide ((*)NO) is a vasodilator and polyphenolic compounds induce endothelium-dependent vasorelaxation in vitro. 2. Here we studied whether a diet rich in dealcoholated red wine (DRW) increases acetylcholine-induced vasorelaxation and whether ingestion of DRW-, quercetin- or catechin-rich diets modifies the (*)NO-cyclic guanosine-3',5'-monophosphate (cyclic GMP) pathway and superoxide anion (O2(.-)) release in aorta in a resting state in rats fed semi-purified diets containing either 35% (v w(-1)) DRW, 0.3% (w w(-1)) quercetin or 0.3% (w w(-1)) catechin for 10 days. 3. (*)NO-mediated vasorelaxation induced by acetylcholine was greater in rats fed the DRW-rich diet than in those that received the control diet. 4. Expression of endothelial (*)NO synthase (eNOS) was similar in the four dietary groups. The aortic rings of rats fed either the DRW-, quercetin-, or catechin rich diets showed higher NOS activity, (*)NO production and cyclic GMP content than those of rats fed the control diet. No changes were observed in O2(.-) production. 5. In summary, diets rich in either DRW, quercetin or catechin induced endothelium-dependent vasorelaxation in rat aorta in a resting state through the enhancement of (*)NO production, without modifying O2(.-) generation, thus the bioavailability of (*)NO was increased. The increase in the (*)NO-cyclic GMP pathway explains the beneficial effect of flavonoids at vascular level. PMID- 11861313 TI - TNF ligands and receptors--a matter of life and death. PMID- 11861319 TI - Influence of antioxidant depletion on nitrergic relaxation in the pig gastric fundus. AB - 1. The hypothesis that endogenous tissue antioxidants might explain the inability of the superoxide generators 6-anilino-5,8-quinolinedione (LY83583) and hydroquinone (HQ) and of the NO-scavengers hydroxocobalamin (HC) and 2-(4 carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (c-PTIO) to affect nitrergic neurotransmission in the porcine gastric fundus was tested by selective pharmacological depletion of respectively Cu/Zn superoxide dismutase (Cu/Zn SOD) and reduced glutathione (GSH) in circular smooth muscle preparations. 2. Diethyldithiocarbamate (DETCA; 3x10(-3) M), which almost completely abolished tissue Cu/Zn SOD activity, had no effect per se on nitrergic relaxations induced by either electrical field stimulation (EFS; 4 Hz, 10 s) or exogenous nitric oxide (NO; 10(-5) M). In these DETCA-treated tissues however, electrically induced nitrergic relaxations became sensitive to inhibition by LY83583 (10(-5) M) or HC (10(-4) M), but not by HQ (10(-4) M) or c-PTIO (10(-4) M); only for the combination of DETCA plus LY83583, this inhibition was partially reversed by exogenous Cu/Zn SOD (1000 u ml(-1)). 3. Immunohistochemical analysis of porcine gastric fundus revealed a 100% colocalization of Cu/Zn SOD and neuronal nitric oxide synthase (nNOS) in the intrinsic neurons of the myenteric plexus. 4. Buthionine sulphoximine (BSO; 10(-3) M) in the absence or presence of LY83583 (10(-5) M) or HC (10(-4) M) did not alter nitrergic relaxations, although it reduced per se the tissue GSH content to 62% of control. 5. Pharmacological depletion studies, corroborated by immunohistochemical data, thus suggest a role for Cu/Zn SOD but not for GSH in nitrergic neurotransmission in the porcine gastric fundus. PMID- 11861320 TI - Prevention of a hypoxic Ca(2+)(i) response by SERCA inhibitors in cerebral arterioles. AB - 1. The aim of the study was to investigate the mechanism of a novel effect of hypoxia on intracellular Ca(2+) signalling in rabbit cerebral arteriolar smooth muscle cells, an effect that was resistant to the L-type Ca(2+) channel antagonist methoxyverapamil (D600). 2.[Ca(2+)](i) of smooth muscle cells in intact arteriolar fragments was measured using the Ca(2+)-indicator dye fura-PE3. Hypoxia (PO(2) 10 - 20 mmHg) lowered basal [Ca(2+)](i) but did not inhibit Ca(2+) entry pathways measured by Mn(2+)-quenching of fura-PE3. 3. The effect of hypoxia was completely prevented by thapsigargin or cyclopiazonic acid, selective inhibitors of sarcoplasmic reticulum Ca(2+) ATPase (SERCA). Since these inhibitors do not block Ca(2+) extrusion or uptake via the plasma membrane, the data indicate that the effect of hypoxia depends on a functional sarcoplasmic reticulum. 4. Because actions of nitric oxide (NO) on vascular smooth muscle are also prevented by SERCA inhibitors it was explored whether the effect of hypoxia occurred via modulation of endogenous NO release. Residual NOS-I and NOS-III were detected by immunostaining, and there were NO-dependent effects of NOS inhibitors on Ca(2+)(i)-signalling. Nevertheless, inhibition of endogenous NO production did not prevent the effect of hypoxia on [Ca(2+)](i). 5. The experiments reveal a novel nitric oxide-independent effect of hypoxia that is prevented by SERCA inhibitors. PMID- 11861321 TI - 5-HT(1B) but not 5-HT(6) or 5-HT(7) receptors mediate depression of spinal nociceptive reflexes in vitro. AB - 1. The identity of the serotonin (5-HT) receptors modulating the transmission of segmental C-fibre mediated signals was studied using an in vitro preparation of the hemisected spinal cord from rat pups. 2. Responses to trains of stimuli delivered to a lumbar dorsal root were recorded from the corresponding ventral root. The resulting cumulative depolarization (CD) mediated by unmyelinated fibres was quantified in terms of integrated area. The amplitude of the mono synaptic reflex was also measured. Serotonergic agents were superfused at known concentrations and their effects on the reflexes evaluated. 3. 5-HT had depressant effects on the CD (EC(50) 34 microM). The rank order of potency of agonists for the depression of the CD was 5-carboxamidotryptamine (5-CT)>alpha methylserotonin (alpha-met-5-HT) approximately 5-HT>42-methylserotonin (2-met-5 HT)approximately 8-OH-DPAT. 4. All the agonists including 2-met-5-HT and 8-OH DPAT had strong depressant effects on the mono-synaptic reflex with the following order of potency: 5-CT>48-OH-DPAT>4alpha-met-5-HT approximate5-HTapproximate2-met 5-HT. 5. The inhibitory effects of 5-HT, alpha-met-5-HT and 5-CT were attenuated by the non-specific 5-HT antagonist methiothepin (1 microM) and by the 5 HT(1A/1B) antagonist SDZ 21009 (100 nM) but not by the selective 5-HT(1A) antagonist WAY 100135 (1 microM). 6. Other antagonists known to block 5-HT(2), 5 HT(6) and/or 5-HT(7) receptors (ketanserin, RO 04-6790, ritanserin and clozapine) did not change the effect of the agonists. 7. The data suggest an important contribution of 5-HT(1B) receptors to the inhibition of spinal C-fibre mediated nociceptive reflexes but no experimental support was found for the intervention of 5-HT(2), 5-HT(6) or 5-HT(7) receptors in this in vitro model. PMID- 11861322 TI - Orphanin FQ inhibits capsaicin-induced thermal nociception in monkeys by activation of peripheral ORL1 receptors. AB - 1. Orphanin FQ (OFQ), an endogenous peptide for ORL1 receptors, has been identified. Although the actions of OFQ have much in common with those of opioid peptides at the cellular level, behavioral studies in rodents seem conflicting. 2. The aim of this study was to investigate the potential pronociceptive or antinociceptive function of peripheral ORL1 receptors in primates. Experiments were conducted to verify whether local administration of OFQ can attenuate capsaicin-induced nociception and whether peripheral ORL1 receptors selectively mediate the local action of OFQ in monkeys. 3. Capsaicin (100 microg) was administered subcutaneously in the tail to locally evoke a nociceptive response (thermal allodynia/hyperalgesia), which was manifested as a reduced tail withdrawal latency in normally innocuous 46 degreeC warm water. 4. Co administration of OFQ (1--30 microg) with capsaicin in the tail dose-dependently inhibited thermal nociception. However, a locally effective dose of OFQ (30 microg), when applied in the back, did not inhibit capsaicin-induced nociception. 5. OFQ-induced local antinociception was antagonized by a small dose (10 microg) of J-113397, a selective ORL1 receptor antagonist, in the tail. Similarly, s.c. administration of 10 microg of J-113397 in the back did not antagonize local antinociception of OFQ. 6. In addition, s.c. administration of either OFQ or J 113397 in the tail alone did not change its thermal nociceptive threshold. Local administration of opioid receptor antagonists selective for mu, kappa, and delta opioid receptors did not antagonize OFQ-induced local antinociception. Local administration of J-113397 also did not interfere with the local actions of mu, kappa, and delta opioid agonists in the tail. 7. These results provide the first functional evidence that activation of peripheral ORL1 receptors produces thermal antinociception in primates and this action is independent of antinociception produced at classical opioid receptors. PMID- 11861323 TI - Stimulation of beta(3)-adrenoceptors causes phosphorylation of p38 mitogen activated protein kinase via a stimulatory G protein-dependent pathway in 3T3-L1 adipocytes. AB - 1. This study deals with phosphorylation and activation of p38 mitogen-activated protein kinase (MAPK) via beta(3)-adrenoceptor (AR) and the signal transduction pathway in 3T3-L1 adipocytes. 2. beta(3)-AR agonist BRL37344A (10 nM) caused phosphorylation and activation of p38 MAPK in 3T3-L1 adipocytes but not in fibroblasts. BRL37344A and also the other beta(3)-AR agonists, CGP12177A and SR58611A, caused p38 MAPK phosphorylation in dose-dependent manners. 3. The p38 MAPK phosphorylations by BRL37344A (10 nM), CGP12177A (100 nM), and SR58611A (10 nM) were not antagonized by beta(1)- and beta(2)-ARs antagonist 1-propranolol (100 nM) but blocked by beta(3)-AR antagonist SR59230A (10 microM), suggesting the phosphorylation was caused via beta(3)-AR. 4. The phosphorylations of p38 MAPK were completely abolished by treatment with cholera toxin (CTX) but not pertussis toxin (100 ng ml(-1), 24 h). Activation of Gs by CTX (100 ng ml(-1)) and adenylyl cyclase by forskolin mimicked p38 MAPK phosphorylation. 5. p38 MAPK phosphorylation by BRL37344A was reduced to almost 50% by cyclic AMP-dependent protein kinase (PKA) inhibitors such as H89 (10 microM) and PKI (10 microM). A src-family tyrosine kinases inhibitor PP2 (1 microM) also halved the p38 MAPK phosphorylation. Combined use of H89 (10 microM) and PP2 (10 microM) did not bring about further inhibition. 6. These results suggest that beta(3)-AR caused phosphorylation of p38 MAPK via Gs protein and partly through a pathway involving PKA and src-family kinase(s), although the contribution of the unidentified pathway remains to be clarified. PMID- 11861324 TI - Attenuation of cardiac fibrosis by pirfenidone and amiloride in DOCA-salt hypertensive rats. AB - 1. This study has administered pirfenidone (5-methyl-1-phenyl-2-[1H]-pyridone) or amiloride to attenuate the remodelling and associated functional changes, especially an increased cardiac stiffness, in DOCA-salt hypertensive rats. 2. In control rats, the elimination half-life of pirfenidone following a single intravenous dose of 200 mg kg(-1) was 37 min while oral bioavailability at this dose was 25.7%. Plasma pirfenidone concentrations in control rats averaged 1.9 +/ 0.1 microg ml(-1) over 24 h after 14 days' administration as a 0.4% mixture in food. 3. Pirfenidone (approximately 250-300 mg kg(-1) day(-1) as 0.4% in food) and amiloride (1 mg kg(-1) day(-1) sc) were administered for 2 weeks starting 2 weeks post-surgery. Pirfenidone but not amiloride attenuated ventricular hypertrophy (2.69 +/- 0.09, UNX 2.01 +/- 0.05. DOCA-salt 3.11 +/- 0.09 mg kg(-1) body wt) without lowering systolic blood pressure. 4. Collagen deposition was significantly increased in the interstitium after 2 weeks and further increased with scarring of the left ventricle after 4 weeks; pirfenidone and amiloride reversed the increases and prevented further increases. This accumulation of collagen was accompanied by an increase in diastolic stiffness constant; both amiloride and pirfenidone reversed this increase. 5. Noradrenaline potency (positive chronotropy) was decreased in right atria (neg log EC50: control 6.92 +/- 0.06; DOCA-salt 6.64 +/- 0.08); pirfenidone but not amiloride reversed this change. Noradrenaline was a more potent vasoconstrictor in thoracic aortic rings (neg log EC50: control 6.91 +/- 0.10; DOCA-salt 7.90 +/- 0.07); pirfenidone treatment did not change noradrenaline potency. 6. Thus, pirfenidone and amiloride reverse and prevent cardiac remodelling and the increased cardiac stiffness without reversing the increased vascular responses to noradrenaline. PMID- 11861325 TI - Specific [(3)H]-guanosine binding sites in rat brain membranes. AB - 1. Extracellular guanosine has diverse effects on many cellular components of the central nervous system, some of which may be related to its uptake into cells and others to its ability to release adenine-based purines from cells. Yet other effects of extracellular guanosine are compatible with an action on G-protein linked cell membrane receptors. 2. Specific binding sites for [(3)H]-guanosine were detected on membrane preparations from rat brain. The kinetics of [(3)H] guanosine binding to membranes was described by rate constants of association and dissociation of 2.6122 x 10(7) M(-1) min(-1) and 1.69 min(-1), respectively. A single high affinity binding site for [(3)H]-guanosine with a K(D) of 95.4 +/- 11.9 nM and B(max) of 0.57 +/- 0.03 pmol mg(-1) protein was shown. This site was specific for guanosine, and the order of potency in displacing 50 nM [(3)H] guanosine was: guanosine=6-thio-guanosine > inosine > 6-thio-guanine > guanine. Other naturally occurring purines, such as adenosine, hypoxanthine, xanthine caffeine, theophylline, GDP, GMP and ATP were unable to significantly displace the radiolabelled guanosine. Thus, this binding site is distinct from the well characterized receptors for adenosine and purines. 5. The addition of GTP produced a small concentration-dependent decrease in guanosine binding, suggesting this guanosine binding site was linked to a G-protein. 6. Our results therefore are consistent with the existence of a novel cell membrane receptor site, specific for guanosine. PMID- 11861326 TI - Cyclosporin A-induced free radical generation is not mediated by cytochrome P 450. AB - 1. Reactive oxygen species (ROS) have been proposed to play a role in the side effects of the immunosuppressive drug cyclosporin A (CsA). 2. The aim of this study was to investigate whether cytochrome P-450 (CYP) dependent metabolism of CsA could be responsible for ROS generation since it has been suggested that CsA may influence the CYP system to produce ROS. 3. We show that CsA (1 -- 10 microM) generated antioxidant-inhibitable ROS in rat aortic smooth muscle cells (RASMC) using the fluorescent probe 2,7-dichlorofluorescin diacetate. 4. Using cytochrome c as substrate, we show that CsA (10 microM) did not inhibit NADPH cytochrome P 450 reductase in microsomes prepared from rat liver, kidney or RASMC. 5. CsA (10 microM) did not uncouple the electron flow from NADPH via NADPH cytochrome P-450 reductase to the CYP enzymes because CsA did not inhibit the metabolism of substrates selective for several CYP enzymes that do not metabolize CsA in rat liver microsomes. 6. CsA (10 microM) did not generate more radicals in CYP 3A4 expressing immortalized human liver epithelial cells (T5-3A4 cells) than in control cells that do not express CYP 3A4. 7. Neither diphenylene iodonium nor the CYP 3A inhibitor ketoconazole were able to block ROS formation in rat aortic smooth muscle or T5-3A4 cells. 8. These results demonstrate that CYP enzymes do not contribute to CsA-induced ROS formation and that CsA neither inhibits NADPH cytochrome P-450 reductase nor the electron transfer to the CYP enzymes. PMID- 11861327 TI - Glucocorticoids reverse IL-1beta-induced impairment of beta-adrenoceptor-mediated relaxation and up-regulation of G-protein-coupled receptor kinases. AB - 1. The aim of the present study was to examine the effects of glucocorticoid dexamethasone on airway responsiveness to albuterol after intratracheal instillation of saline or IL-1beta in Brown-Norway rats in vivo and to elucidate the molecular mechanism of this effect. 2. IL-1beta caused a significant reduction in albuterol-mediated relaxation to protect against MCh-induced bronchoconstriction. Dexamethasone attenuated the IL-1beta-induced impaired relaxation while alone had no effect when compared to rats treated identically with saline. 3. The density of beta(2)-adrenoceptors was significantly reduced in lung membranes harvested from IL-1beta-treated rats, which was associated with impaired isoproterenol- and forskolin-stimulated cyclic AMP accumulation and adenylyl cyclase (AC) activity ex vivo. Dexamethasone did not prevent IL-1beta induced down-regulation of beta(2)-adrenoceptors but completely blocked IL-1beta induced impairment of cyclic AMP accumulation and AC activity stimulated by isoproterenol and forskolin. 4. The inhibitory G-protein subtypes, G(ialpha1), G(ialpha2) and G(ialpha3), were detected in lung membranes prepared from all groups of rats but the intensity of G(ialpha1) and G(ialpha2) was markedly increased in IL-1beta-treated rats, which were not prevented by dexamethasone. 5. The activity of cytosolic GRK and the expression of GRK2 and GRK5 were elevated in the lung of IL-1beta-treated rats, which were completely abolished by dexamethasone. 6. These results indicate that treatment of rats with IL-1beta results in desensitization of pulmonary beta(2)-adrenoceptors. In light of data obtained in this study, we propose that both the decrease in AC activity and the increase in GRK activity, which are reversed by dexamethasone, may underlie beta(2)-adrenoceptor desensitization. PMID- 11861328 TI - Constitutive activation of the neurotensin receptor 1 by mutation of Phe(358) in Helix seven. AB - 1. The neurotensin receptor 1, NTS1, is a G protein-coupled receptor with seven transmembrane domains (TM) that mediates most of the known effects of the neuropeptide. Our previous studies have pointed to extracellular loop 3 and adjacent TM7 as being potentially involved in agonist-induced activation of the NTS1. 2. Here we investigated residues in these domains that might be involved in transconformational activation of the rat NTS1. Single amino acid mutated receptors were expressed in COS cells and inositol phosphate (IP) and cyclic AMP productions were studied. 3. The F358A mutation in TM7 resulted in a time- and receptor concentration-dependent increase in spontaneous IP production. At expression levels of 12 pmol mg(-1), agonist-independent IP production was increased 10 fold over basal for the F358A mutant receptor whereas the wild type NTS1 exhibited virtually no spontaneous activity at expression levels of 7.5 pmol mg(-1). 4. Neurotensin remained agonist on the F358A mutant receptor with a maximal effect that amounted to greater than twice basal IP levels. SR 48692 was inverse agonist at the mutant receptor, reversing IP production almost back to the levels measured in wild type NTS1-transfected cells. 5. Cyclic AMP production was not constitutively activated with the F358A mutant receptor but was stimulated by neurotensin with the same concentration dependence as that observed with the wild type NTS1. 6. This is the first report, to our knowledge, of a constitutively active mutant of the NTS1. The data are consistent with TM7 being involved in the transconformational changes that lead to agonist-induced coupling of the NTS1 to Gq. PMID- 11861329 TI - Proarrhythmic potential of halofantrine, terfenadine and clofilium in a modified in vivo model of torsade de pointes. AB - 1. This study was designed to compare the proarrhythmic activity of the antimalarial drug, halofantrine and the antihistamine, terfenadine, with that of clofilium a K(+) channel blocking drug that can induce torsade de pointes. 2. Experiments were performed in pentobarbitone-anaesthetized, open-chest rabbits. Each rabbit received intermittent, rising dose i.v. infusions of the alpha adrenoceptor agonist phenylephrine. During these infusions rabbits also received increasing i.v. doses of clofilium (20, 60 and 200 nmol kg(-1) min(-1)), terfenadine (75, 250 and 750 nmol kg(-1) min(-1)), halofantrine (6, 20 and 60 micromol kg(-1)) or vehicle. 3. Clofilium and halofantrine caused dose-dependent increases in the rate-corrected QT interval (QTc), whereas terfenadine prolonged PR and QRS intervals rather than prolonging cardiac repolarization. Progressive bradycardia occurred in all groups. After administration of the highest dose of each drug halofantrine caused a modest decrease in blood pressure, but terfenadine had profound hypotensive effects resulting in death of most rabbits. 4. The total number of ventricular premature beats was highest in the clofilium group. Torsade de pointes occurred in 6 out of 8 clofilium-treated rabbits and 4 out of 6 of those which received halofantrine, but was not seen in any of the seven terfenadine-treated rabbits. 5. These results show that, like clofilium, halofantrine can cause torsade de pointes in a modified anaesthetized rabbit model whereas the primary adverse effect of terfenadine was cardiac contractile failure. PMID- 11861330 TI - Inhibition of slow Ca(2+)-activated K(+) current by 4-aminopyridine in rat hippocampal CA1 pyramidal neurones. AB - 1. The effect of 4-aminopyridine (4-AP) on the slow afterhyperpolarization (sAHP) seen after high frequency dendritic or somatic firing was investigated in rat hippocampal CA1 pyramidal neurones (PC). Intracellular recordings were obtained from the distal apical dendrites and somata and suprathreshold depolarizing current pulses were used to evoke a sAHP. The sAHP was blocked by low concentrations of carbacholine (Cch) but insensitive to high concentrations of apamin. 2. In the presence of extracellular 4-AP, the first dendritic sAHP evoked was reduced compared to a maximal sAHP evoked in the absence of 4-AP. The reduction was evident at submillimolar concentration and increased to about 80% with 4 mM 4-AP. 3. The stability of the 4-AP-induced block was affected by the type of anion used in the electrode solution. With K(+) acetate (KAc) or K(+) methylsulphate (KMeSO(4)) containing electrodes, the block was progressively removed during the initial 300 - 400 s of recordings. With KCl containing electrodes, the block remained stable and was 10% larger than that obtained with acetate. Detailed investigations showed that intracellular acetate promotes the removal of the 4-AP-induced block in an activity-dependent manner. 4. Intracellularly applied 4-AP also induced an acetate-sensitive block of the dendritic sAHP. 5. 4-AP also blocked the somatic sAHP and the stability of the block showed the same sensitivity towards anions as the dendritic sAHP. 6. Thus 4 AP appears to block the slow Ca(2+)-activated K(+) current underlying the sAHP in a complex manner which is sensitive to certain types of anions. PMID- 11861331 TI - Effects of ketanserin and DOI on spontaneous and 5-HT-evoked peristalsis of the pig ureter in vivo. AB - 1. The influence of 5-hydroxytryptamine (5-HT) receptor agonists and antagonists on the ureter motility was investigated in vivo on intact ureters of anaesthetized pigs. Drugs were administered intravenously or topically. 2. 5-HT induced a dose-dependent increase in the frequency of ureter contractions in anaesthetized pigs when given intravenously (0.0001-1 mg kg(-1); ED(50) 0.066 mg kg(-1)) or topically (0.001-1 mg ml(-1); EC(50) 0.043 mg ml(-1)). Significant increases in heart rate and blood pressure were observed when the drug was given intravenously but not topically. 3. The 5-HT(2A) agonist, DOI (1-(2,5-dimethoxy-4 iodophenyl)-2-aminopropane) increased the frequency of ureteral contractions in a dose-dependent manner (1-300 microg kg(-1) i.v.). Calculation of ED(50) indicated this compound to be about 1.5 times more potent with an efficacy of 23% compared to 5-HT. 4. The 5-HT(2A/2C) antagonist, ketanserin (0.5 mg kg(-1)) and the 5 HT(2C) antagonist, methysergide (1 mg kg(-1)) antagonized the 5-HT-induced ureter peristalsis when given intravenously. Contraction amplitude, blood pressure and heart rate were not affected by the antagonists. 5. Intravenous (0.0001-1 mg kg( 1)) and topical (0.0001-1 microg ml(-1)) ketanserin significantly decreased the frequency of spontaneous ureteral contractions to about 30% of controls, which could be partly reversed by 5-HT (0.3 mg kg(-1) i.v.). The contraction amplitude, contractions of the contralateral, saline perfused ureter, heart rate and mean arterial blood pressure were not affected. 6. Thus, contractility of porcine ureter is mediated by 5-HT(2) receptors. Their antagonists ketanserin and methysergide seem to be promising drugs for treatment of acute ureteric colic or in preparing the ureter for ureteroscopy. PMID- 11861332 TI - Pharmacological characterization of the chemokine receptor, CCR5. AB - 1. We investigated the effects of a number of naturally occurring chemokines (MIP 1alpha, MIP-1beta, RANTES, MCP-2, MCP-3, MCP-4) on different processes linked to the chemokine receptor CCR5 in recombinant CHO cells expressing the receptor at different levels. 2. Internalization of CCR5 following chemokine treatment was studied and MIP-1alpha, MIP-1beta and RANTES (50 nM) were able to induce internalization (similar50%) of the receptor. Internalization due to MCP-2, MCP-3 and MCP-4 was less (similar20%). 3. Phosphorylation of CCR5 following chemokine treatment was studied and MIP-1alpha, MIP-1beta and RANTES (50 nM) were able to induce phosphorylation of CCR5 whereas the other chemokines did not induce CCR5 phosphorylation. 4. MIP-1alpha, MIP-1beta, RANTES and MCP-2 were able to stimulate [(35)S]-GTPgammaS binding, an index of receptor/G protein activation, whereas MCP-3 and MCP-4 had no effect in this assay. MCP-2 was a partial agonist (similar80%) compared to MIP-1alpha, MIP-1beta and RANTES, which gave similar maximal stimulations in this assay. 5. MIP-1alpha, MIP-1beta, RANTES, MCP-2 and MCP-4 were able to stimulate increases in intracellular calcium ions via activation of CCR5 whereas MCP-3 was without effect. 6. It is concluded that different chemokines interacting with CCR5 mediate different patterns of cellular responses. PMID- 11861333 TI - Divergent effects of vitamin C on relaxations of rabbit aortic rings to acetylcholine and NO-donors. AB - 1. Vitamin C may influence NO-dependent relaxation independently of effects on oxidant stress. 2. We investigated effects of vitamin C (0.1 -- 10 mmol l(-1)) on relaxation of pre-constricted rabbit aortic rings to acetylcholine (ACh), authentic NO and the NO-donors glyceryl trinitrate (GTN), nitroprusside (NP) and S-nitroso-N-acetyl-penicillamine (SNAP). DETCA (2 -- 6 mmol l(-1)), a cell permeable inhibitor of endogenous Cu-Zn superoxide dismutase (SOD) was used to increase intracellular superoxide anion (O(2)(-)). 3. Vitamin C reduced the response to ACh (71 +/- 7% inhibition of maximum relaxation at 10 mmol l(-1)) and inhibited relaxation to authentic NO. Vitamin C inhibited relaxation to GTN but potentiated relaxations to NP and SNAP, causing a parallel shift to a lower concentration range of the log dose-response curve by approximately one log unit at the highest dose. 4. Vitamin C increased the concentration of NO in bath solution (plus EDTA, 1.0 mmol l(-1)) following the addition of SNAP from 53 +/- 14 to 771 +/- 101 nmol l(-1) over the range 0.1-3.0 mmol l(-1). 5. DETCA inhibited relaxation to ACh (71 +/- 9% inhibition of maximum relaxation). This inhibition was abolished by a cell permeable SOD mimetic, but not by vitamin C. DETCA inhibited relaxation to SNAP but not that to NP nor to GTN. 6. Vitamin C inhibits endothelium-dependent relaxations of rabbit aortic rings to ACh and authentic NO and does not reverse impaired relaxation resulting from increased intracellular oxidant stress. Vitamin C potentiates relaxation to the NO-donors NP and SNAP by a mechanism that could involve release of NO from nitrosothiols. PMID- 11861334 TI - Effects of chronic drug treatments on increases in intracellular calcium mediated by nicotinic acetylcholine receptors in SH-SY5Y cells. AB - 1. SH-SY5Y cells express alpha7 and alpha3* subtypes of nicotinic acetylcholine receptors (AChR). Numbers of these receptors are upregulated by chronic treatment with nicotinic agonists or KCl. In this study we have examined the functional consequences of these drug treatments on nicotine- or KCl-evoked increases in [Ca(2+)](i), in SH-SY5Y cells. 2. In untreated cells, nicotine increased [Ca(2+)](i) (EC(50) 7.5 microM). Responses to 10 microM nicotine were abolished by the non-selective nicotinic antagonist mecamylamine and were partially blocked by alpha7-selective antagonists, the alpha3beta2*-selective antagonist alpha conotoxin-MII, and by cadmium and verapamil. 3. After treatment for 4 days with nicotinic agonists, nicotine-evoked increases in [Ca(2+)](i) were significantly decreased by about 25%. Nicotine-evoked responses were paradoxically increased in the presence of acute methyllycaconitine (MLA; an alpha7-selective antagonist) although other alpha7-selective antagonists were without effect, while alpha conotoxin-MII gave a partial inhibition. The increase observed with MLA was abolished by mecamylamine but not by alpha-conotoxin-MII and was still observed 24 h after chronic nicotine treatment. 4. After treatment for 4 days with KCl, nicotine-evoked increases in [Ca(2+)](i) were also decreased by 25%, but acute MLA was without effect. Responses to 20 mM KCl were unchanged by prior treatment with nicotine or KCl. Treatment for 4 days with 5 microM verapamil reduced responses to both nicotine and KCl by about 50%. 5. Multiple nicotinic AChR subtypes contribute to nicotine-evoked increases in [Ca(2+)](i) in SH-SY5Y cells. Responses to acute nicotine are reduced after chronic nicotine or KCl treatment, with loss of the component attributed to the alpha7 subtype. However, in nicotine treated cells this effect is reversed when nicotine stimulation is applied in the presence of acute MLA. The antagonist may assist in converting a non-functional alpha7 nicotinic AChR to a conducting state. PMID- 11861335 TI - Pulmonary hypertension secondary to left ventricular dysfunction: the role of nitric oxide and endothelin-1 in the control of pulmonary vascular tone. AB - 1. Using an in vivo model of pulmonary hypertension (PHT) secondary to left ventricular dysfunction (LVD), the pulmonary arterial response to the nitric oxide synthase (NOS) blocker L-NAME (30 micromol.min(-1) i.v.) and the subsequent responses to cumulatively administered endothelin-1 (ET-1) (0.001 -- 4 nmol.kg( 1) i.v.) or big ET-1 (0.1 -- 2.0 nmol.kg(-1) i.v.) were studied. Additionally, the effect of the non-selective ET-1 receptor antagonist, SB209670, was investigated. 2. Eight weeks after coronary artery ligation or sham operation, rabbits demonstrated increased mean pulmonary arterial pressure (PAP) accompanied by right ventricular hypertrophy. 3. Blockade of NOS caused a greater increase in basal PAP (increased by 7.7 +/- 1.1 mmHg c.f. 3.8 +/- 1.0 mmHg in controls, P<0.05) and uncovered a greater pulmonary pressor response to exogenous ET-1 in rabbits with PHT (increased by 10.2 +/- 2.3 mmHg c.f. 4.9 +/- 1.0 mmHg in controls, P<0.05). 4. Big ET-1 evoked a pulmonary pressor effect, in both groups of rabbits, that was increased following blockade of NOS and was more potent in rabbits with PHT. 5. The non-selective ET-1 receptor antagonist, SB209670, reduced basal PAP (from 16.9 mmHg to 15.9 mmHg, P < 0.05) in rabbits with PHT and blocked the response to ET-1 in the presence of L-NAME. 6. In conclusion, the results demonstrate that basal NO activity masks a pulmonary pressor response to exogenously administered ET-1. An increased responsiveness to ET-1 was shown in the pulmonary arterial bed of rabbits with PHT secondary to LVD, implicating a pathophysiological role for ET-1 in this model. PMID- 11861337 TI - Single-lung ventilation in pediatrics. PMID- 11861336 TI - Involvement of caspases and calpains in cerebrocortical neuronal cell death is stimulus-dependent. AB - 1. Caspases and calpains are mediators of apoptotic cell death. The objective of this study was to determine the role of caspases and calpains in primary cerebrocortical neuronal (CCN) death in response to a range of stimuli which reportedly induce neuronal apoptosis. 2. Cell death of primary cultures of rat CCN was induced by staurosporine (STS), C2-ceramide (CER), camptothecin (CMT), hydrogen peroxide (H(2)O(2)) or N-methyl-D-aspartate (NMDA). Caspase and calpain activity were assessed by cleavage of alpha-fodrin or fluorogenic substrates. 3. Cell death was analysed by lactate dehydrogenase (LDH) assay in the absence or presence of the pan-caspase inhibitor Boc-Asp-(OMe)-Fluoromethylketone (Baf) and/or the calpain inhibitor calpeptin (CP). Cell death induced by STS, CER or CMT was accompanied by chromatin condensation and activation of multiple caspases, particularly caspase-3-type proteases. Hydrogen peroxide (H(2)O(2)) treatment was accompanied by activation of caspases -1, -6 and -8, but not -3, whereas none of the caspases tested were activated in response to NMDA. 4. With the exception of H(2)O(2), when cell death was accompanied by caspase activation, it was significantly suppressed by Baf. 5. All stimuli also induced calpain activation, but calpeptin only suppressed cell death induced by H(2)O(2). Furthermore, co-treatment with Baf and calpeptin did not alter the cell death relative to either inhibitor alone. 6. These findings suggest the existence of stimulus-dependent routes for the activation of caspases and calpains during death of cortical neurones and imply that although caspases and calpains are activated, their involvement in the execution of cell death varies with the stimulus. PMID- 11861338 TI - Pulmonary artery catheters in cardiovascular surgery. PMID- 11861339 TI - The inter-rater and intra-rater reliability of a new Canadian oral examination format in anesthesia is fair to good. AB - PURPOSE: In response to the Royal College's request to improve the validity and reliability of oral examinations, the Examination Board in anesthesia proposed a structured oral examination format. Prior to its introduction, we studied this format in two residency programs to determine reliability of the examiners. METHODS: Twenty faculty and 26 residents from two Canadian residency programs participated (Sites A and B). Pairs of examiners scored five or six residents examined consecutively on two standardized questions using global rating scales with anchored performance criteria. Residents' performances were scored independently during the examination (Time 1) and later from a videotaped recording (Time 2). Correlations between scores of the pairs of examiners and between scores of each examiner were determined. RESULTS: Correlations demonstrating inter-rater agreement between examiners at Site A ranged from -.324 to.915 (mean.506) at Time 1. At Time 2, correlations ranged from.64 to.887 (mean.791). At Site B correlations ranged from.279 to.989 (mean.707) at Time 1 and at Time 2 correlations ranged from -.271 to.924 (mean.477). Correlations demonstrating intra-rater agreement of examiners at Site A ranged from.054 to.983 (mean.723) and at Site B correlations ranged from -.055 to.974 (mean.662). Correlations >0.4 were seen in 80% of the scores and >0.7 in 50% indicating fair to good intra-rater and inter-rater reliability using this format. CONCLUSIONS: Despite the limitations of our study our results compare favourably with those previously reported in anesthesia. We recommend the adoption of this format to the Royal College of Physicians and Surgeons of Canada Examination Board. PMID- 11861340 TI - Apfel's simplified score may favourably predict the risk of postoperative nausea and vomiting. AB - PURPOSE: To compare two of the latest published scores for predicting postoperative nausea and vomiting (PONV) in potentially high-risk patients. METHODS: Adult in-patients scheduled for throat, thyroid, breast or gynecological surgery under general inhalational anesthesia were studied prospectively over 24 hr for PONV. The latest published score considers four risk factors: female gender, previous history of PONV or motion sickness, non-smoking status and postoperative use of opioids (Apfel-score). The previously published score includes, in addition to these factors, duration, type of anesthesia and surgery (Sinclair-score). The two scores were compared by calculating the area under a receiver operating characteristic (ROC)-curve and plotting calibration curves of the predicted and the observed incidence of PONV. RESULTS: Five hundred consecutive patients were studied and patients who received prophylactic antiemetics were excluded. Of the remaining 428 patients 49.5% suffered from PONV. Multivariable analysis revealed that age, gender, previous history of PONV or motion sickness and postoperative use of opioids had an impact on PONV. The area under the ROC-curve was significantly greater for the Apfel-score compared to the Sinclair-score (0.71 vs 0.64, P=0.008). The correlation between the predicted (x) and the observed (y) incidence for the Apfel-score and for the Sinclair-score was y=1.08x - 0.07 and y=0.93x + 0.27. CONCLUSION: In our hospital, the simplified Apfel-score presented with favourable discriminating and calibration properties for predicting the risk of PONV. Therefore, we have implemented this score in our daily clinical practice as well as in an ongoing antiemetic trial. PMID- 11861341 TI - The HemoCue, a point of care B-hemoglobin photometer, measures hemoglobin concentrations accurately when mixed in vitro with canine plasma and three hemoglobin-based oxygen carriers (HBOC). AB - PURPOSE: Accuracy of measurement of low hemoglobin concentrations using the HemoCue, a B-hemoglobin photometer (HemoCue AB, Angelholm, Sweden) may exhibit significant variability. Infusion of hemoglobin-based oxygen carriers (HBOC) results in low concentrations of plasma hemoglobin. Our study assessed B hemoglobin photometer measurement accuracy of three HBOC: (hemoglobin glutamer 200 (bovine; Oxyglobin, Biopure Corp., Cambridge, MA, USA); hemoglobin glutamer 250 (bovine; Hemopure, Biopure Corp, Cambridge, MA, USA), and hemoglobin raffimer, (human; Hemolink, Hemosol, Inc., Toronto, Ontario, Canada). METHODS: In the laboratory, 45 split canine plasma samples were mixed with hemoglobin glutamer-200 (8.13, 16.25, 32.5 g x L(-1) concentrations), 45 samples were mixed with hemoglobin glutamer-250 (8.13, 16.25, 32.5 g x L(-1) concentrations), 45 with hemoglobin-raffimer (12.5, 25.0, 50.0 g x L(-1) concentrations), and measured. Plasma samples without HBOC served as control. Hemoglobin concentration was determined by a laboratory analyzer (Coulter Corporation, Hiafeah, FL, USA) and B-hemoglobin photometer (HemoCue, Angelholm, Sweden). Two independent technicians performed blinded sample measurements and randomly tested each sample five times. Results were analyzed according to Bland and Altman analysis. RESULTS: B-hemoglobin photometer demonstrated high repeatability for all three HBOCs. Repeatability coefficients were 0.37 g x L(-1) and 0.48 g x L(-1) for hemoglobin glutamer-200, 0.39 g x L(-1) and 0.4 g x L(-1) for hemoglobin glutamer 250 and 1.07 g x L(-1) and 0.85 g x L(-1) for hemoglobin-raffimer. An acceptable agreement was found between the B-hemoglobin photometer and the laboratory analyzer for all three HBOCs tested. CONCLUSION: The B-hemoglobin photometer accurately determined the concentration of three HBOC solutions dissolved in canine plasma. PMID- 11861342 TI - Gender and pain upon movement are associated with the requirements for postoperative patient-controlled iv analgesia: a prospective survey of 2,298 Chinese patients. AB - PURPOSE: To investigate prospectively the influence of patient characteristics upon, and the association of postoperative measurements with, the requirements for postoperative morphine and the assessment of resting pain and pain upon movement in Chinese patients. METHODS: From January 1998 to December 1999, patients receiving patient-controlled iv morphine subsequent to general anesthesia and surgery at our institute (Kaohsiung Veterans General Hospital), were enrolled in the study. Demographic data (such as gender, age, weight, height and education level) and postoperative measurements, including pain scores at rest or during movement, sedation scores and morphine consumption, were recorded. RESULTS: In total 2,298 patients were recruited. Females consumed significantly less morphine via patient-controlled analgesia (PCA) in the first three postoperative days than was the case for males (P <0.05). Gender was the strongest predictor for postoperative morphine requirements. Postoperative pain upon movement was another effective predictor for morphine requirement (P <0.05). Age, body height, body weight, education and operation sites were not associated with morphine consumption. CONCLUSION: Gender and postoperative pain upon movement are the major factors influencing morphine requirement for patient controlled iv morphine analgesia during the first three postoperative days in Chinese patients. PMID- 11861343 TI - Single-lung ventilation in a pediatric patient using a pediatric fibreoptically directed wire-guided endobronchial blocker. AB - PURPOSE: The pediatric wire-guided endobronchial blocker is a new device for single-lung ventilation through small diameter endotracheal tubes. In this case report we will discuss the use of this blocker in a pediatric patient. CLINICAL FEATURES: We successfully placed the pediatric wire-guided endobronchial blocker in a 14-yr-old patient who underwent an aortic coarctation repair. The blocker is a 5-French 70 cm double-lumen catheter. One lumen contains an adjustable wire loop. The other lumen inflates a spherical low pressure, high volume balloon. Through a special bronchoscopy port, the blocker and bronchoscope were placed into a 7.0 cuffed endotracheal tube, the bronchoscope passed through the wire loop of the blocker and advanced towards the left mainstem bronchus. Then the blocker was advanced over the bronchoscope and positioned in the left mainstem bronchus. The balloon was slowly inflated under direct vision and the bronchoscope removed. During the case, single lung ventilation was achieved by inflating the balloon, thus collapsing the lung. At the end of the case, the lung was reinflated by deflating the balloon and the blocker was removed without difficulty. The patient tolerated the procedure well and had an uneventful postoperative course. CONCLUSION: Because of the endobronchial blocker's small diameter, this device can be used in a small endotracheal tube without sacrificing the inner diameter (ID) cross sectional area. Therefore, the patient is ventilated through a conventional endotracheal tube with a larger ID compared to the ID of a double-lumen endotracheal tube. PMID- 11861344 TI - Spinal anesthesia in 62 premature, former-premature or young infants--technical aspects and pitfalls. AB - PURPOSE: To highlight technical aspects and pitfalls of spinal anesthesia (SA) in infants. METHODS: The medical history and perioperative course of all infants who underwent SA over a 28-month period were collected (retrospectively in the first 20). RESULTS: Sixty-two infants underwent surgery under SA. Fifty-five were premature and former-premature, postconceptional age 43.3 +/- 5.0 weeks, weight 3261 +/- 1243 g. Of these, 21 had co-existing disease: cerebral (six), cardiac (nine), pulmonary (11) and urological (six). Hyperbaric tetracaine or bupivacaine 1 mg x kg(-1) with adrenaline was administered. Four infants (three premature) required N(2)O supplementation and three needed general anesthesia. The supplementation rate was similar or lower than in previous studies. Postoperatively, all seven were shown to have lower limb motor and sensory blockade. Complications in premature patients included intraoperative hypoxemia (two), apnea (two) and bradycardia (one). Postoperative complications included bradycardia (three), hypoxemia (one) and apnea and hypoxemia (one). The postoperative complication rate was similar to previous studies. CONCLUSION: Successful SA in infants depends on close attention to preoperative assessment, appropriate patient positioning during and after lumbar puncture, drug dosing and intra- and postoperative cardiorespiratory monitoring. A relatively high dose of hyperbaric solution of tetracaine or bupivacaine with adrenaline should be administered. PMID- 11861345 TI - Active warming of saline or blood is ineffective when standard infusion tubing is used: an experimental study. AB - PURPOSE: To determine the effect of infusion rate, tubing length and fluid composition on the temperature of the infusate reaching the distal end of an infusion tubing with and without active fluid warming. METHODS: Warmed normal saline (W-NS) and packed red blood cells (W-PRBC), were infused with a fluid warmer through a modified infusion set. The fluids were delivered at eight infusion rates from 50 to 999 mL x hr(-1). The infusate temperature was monitored at 20 cm intervals on the iv tubing. The same temperature monitoring protocol was applied to PRBC without warmer (NoW-PRBC). RESULTS: In W-NS and W-PRBC groups, there was a decrease in the infusate temperature, at each flow rate, from the drip chamber to the distal end of tubing ( P <0.001). In NoW-PRBC group, there was a rapid increase in the infusate temperature from the bag to the drip chamber ( P <0.001). Thereafter, there was no change in temperature, except at the 999 mL x hr(-1) infusion rate, where a slight increase in the infusate temperature throughout the tubing was shown. In W-NS and W-PRBC groups increasing the flow rate produced a significant increase in the infusate temperature, at each measurement point ( P <0.001). In the NoW-PRBC group, increasing the flow rate did not alter the infusate temperature. The fluid composition did not influence the infusate temperature. CONCLUSION: There is an important heat exchange within the tubing, which is aggravated at low flow rates. At infusion rates appropriate for pediatric anesthesia the clinical and economic value of fluid warming without the use of heated extension tubing is questionable. PMID- 11861346 TI - The appropriateness of the pulmonary artery catheter in cardiovascular surgery. AB - PURPOSE: The pulmonary artery catheter (PAC) is commonly used in anesthesiology and critical care, but its appropriate (where benefit exceeds risk) application is unknown. This study describes current clinical practice attitudes among anesthesiologists in cardiac and vascular surgery in an effort to determine the most appropriate indications for use of the PAC. METHODS: Anonymous, cross sectional, mailed survey of anesthesiologists in Canada and the USA. Opinions of anesthesiologists about the appropriateness of PAC application were assessed in 36 clinical scenarios, using a nine-point Likert scale. The RAND method was adapted to identify appropriate, inappropriate, and uncertain indications for PAC application. RESULTS: Seventy-seven percent of 345 anesthesiologists responded. They agreed strongly (87%) that use of the PAC is appropriate in patients with severe ventricular impairment and unstable angina. Agreement was also present with ventricular impairment (74%) or unstable angina (55%) alone, but was less strong. A majority (53%) rated the PAC as not appropriate in the routine patient without complicating risk factors. Those who used the PAC more frequently, who had a greater practice volume, and who practised in Canada rated PAC use to be more appropriate in more scenarios. Those who did more continuing medical education rated PAC use to be less appropriate. CONCLUSIONS: While the ideal evaluation of the PAC in clinical practice would be a randomized controlled trial, such an undertaking is time-consuming, expensive, of limited generalizability, and requires clinical equipoise. This study found strong agreement that PAC application is appropriate in some patient scenarios, and agreement that it is inappropriate in others. Description of current practice using this method may identify scenarios where randomized evaluation of the PAC, or other technologies, is likely unnecessary, and others where it is highly likely to be highly beneficial. PMID- 11861347 TI - Ketamine anesthesia for pericardial window in a patient with pericardial tamponade and severe COPD. AB - PURPOSE: To describe the use and concerns of ketamine anesthesia for pericardial window in a patient with pericardial tamponade and severe chronic obstructive pulmonary disease (COPD) with CO(2) retention. CLINICAL FEATURES: A 73-yr-old woman with long-standing COPD and cor pulmonale admitted with pericardial effusion and tamponade had surgery for a pericardial window receiving a total of ketamine 450 mg iv. Arterial pCO(2) increased from 71.8 mmHg preoperatively to 96 mmHg intraoperatively postdrainage of 1000 mL of effusion. Hemodynamic stability and SpO(2) >93% were maintained. Intubation was avoided and concerns of increased pulmonary vascular resistance and potential for right ventricular failure in an already compromised right ventricle were not observed clinically. CONCLUSION: In this patient with pericardial tamponade, COPD and CO(2) retention, the advantages of ketamine included maintaining spontaneous ventilation, avoiding institution and weaning of mechanical ventilation, bronchodilation and relative preservation of the CO(2) response curve. Deleterious effects on right ventricular afterload were not observed. PMID- 11861348 TI - Perioperative use of transesophageal echocardiography by anesthesiologists: impact in noncardiac surgery and in the intensive care unit. AB - BACKGROUND: The American Society of Anesthesiologists (ASA) has published practice guidelines for the use of perioperative transesophageal echocardiography (TEE) but the role and impact of TEE performed by anesthesiologists outside the cardiac operating room (OR) is still poorly explored. We report our experience in the use of TEE in the noncardiac OR, the recovery room and in the intensive care unit (ICU) in a university hospital, and analyze the impact of TEE on clinical decision making. METHODS: Two hundred fourteen patients were included and TEE indications were classified prospectively according to the ASA guidelines. The examinations and data sheets were reviewed by two anesthesiologists with advanced training in TEE. For each examination, it was noted if TEE altered the management according to five groups: 1) changing medical therapy; 2) changing surgical therapy; 3) confirmation of a diagnosis; 4) positioning of an intravascular device; and 5) TEE used as a substitute to a pulmonary artery catheter. RESULTS: Eighty-nine (37%), 67 (31%) and 58 (27%) patients had category I, II and III indications. The impact was more significant in category I where TEE altered therapy 60% of the time compared with 31% and 21% for categories II and III (P < 0.001). The most frequent reason for changing management was a modification in medical therapy in 53 instances (45%). CONCLUSION: Our results confirm a greater impact of TEE performed by anesthesiologists on clinical management for category I compared to category II and III indications in the noncardiac OR surgical setting and in the ICU. PMID- 11861349 TI - Survey of the members of the cardiovascular section of the Canadian Anesthesiologists' Society on the use of perioperative transesophageal echocardiography - a brief report. AB - PURPOSE: Transesophageal echocardiography (TEE) is a useful diagnostic and monitoring tool in the operating room. In the United States, an increasing number of centres are training anesthesiologists to preform intraoperative TEE. In Canada, TEE has been slow to gain acceptance as an intraoperative monitor and little information is available on its use by the anesthesiologists across the country. METHODS: We surveyed all members of the cardiovascular section of the Canadian Anesthesiologists' Society, to find out how many perform TEE, how they acquired their skills and how they use TEE in their practice. RESULTS: The response rate was 48.4%. Most respondents were Canadian-trained cardiac anesthesiologists working in university centres. 91% of respondents stated that their centres offer intraoperative TEE services. Of those services, 35.1% were provided by anesthesiologists only, 13% by cardiologists only, and 51.9% by both. 53.8% of respondents have certification in intraoperative TEE (NBE/SCA, ASE or Provincial College). 90% of respondents use equipment that is less than five years old and multiplane probes are used by almost everyone. There was strong support for Canadian-based continuing medical education events in perioperative TEE. CONCLUSION: TEE appears to be available in most cardiac centres in Canada and anesthesiologists are actively involved in providing intraoperative TEE services, using state-of-the-art equipment. Many anesthesiologists have formal training in TEE. PMID- 11861350 TI - The OxyArm - a new minimal contact oxygen delivery system for mouth or nose breathing. AB - PURPOSE: To describe the development and performance of a new minimal contact oxygen (O(2)) delivery system for both nasal and oral breathing, with capnographic capabilities. METHODS: The development and design challenges of the OxyArm (OA) prototype are described. The innovative design utilizes a headset with a semi-rigid boom and an O(2) diffuser. The OA was compared to the Venturi mask in eight healthy adults after informed consent. Inspired O(2) fractions were measured in the hypopharynx using continuous gas sampling at low to high O(2) flow rates. Mean data were compared using two-tailed paired t tests with significance set at 0.05. RESULTS: The measured inspired O(2) concentration was higher in the OA at 2 (26.3 +/- 2.5 vs 23.3 +/- 0.5, P <0.01) and 6 L x min(-1) (33.5 +/- 3.3 vs 28.8 +/- 1.2, P <0.01) flow rates. At 12 L x min(-1), the O(2) concentration was less in the OA (39.2 +/- 6.3 vs 46.0 +/- 2.7, P <0.02). All subjects found both systems comfortable for the short duration of the study. CONCLUSIONS: The OA delivered predictable concentrations of O(2) over low to medium flow rates. This system is comfortable, easy to use, non-obtrusive, odorless, and latex-free. The ability to monitor capnography makes this device ideal for monitored anesthesia care or in other settings (intensive care) where monitoring of respiration is warranted. This device does not contact the face and thus may be ideal for pediatric patients and those on long-term home O(2) therapy. Further clinical trials in these areas are warranted. PMID- 11861351 TI - Regional blood flows are affected differently by PEEP when the abdomen is open or closed: an experimental rabbit model. AB - OBJECTIVE: The study of induced circulatory changes requires simultaneous assessment of multiple regional circulations because of interactions and compensatory mechanisms. Positive end expiratory pressure mechanical ventilation (PEEP) is known to cause marked, and potentially deleterious, cardiovascular changes. Our aim was to use a comprehensive approach to assess PEEP-induced circulatory changes in open vs closed abdomen animals. MATERIAL AND METHODS: In the anesthetized rabbit, we used implantable Doppler micro-probes to measure blood flow simultaneously in the ascending aorta, inferior vena cava, portal vein, hepatic artery, common carotid artery, and renal artery. We studied spontaneously breathing animals (Group A), and open (Group B) and closed abdomen (Group C) animals mechanically ventilated at 0 (ZEEP) and 12 cm H(2)O PEEP. RESULTS: In Group A, all biological and hemodynamic variables remained unchanged for three hours at the end of the surgical procedure. In Groups B and C, ZEEP produced no significant hemodynamic change. PEEP induced a decrease in carotid, hepatic, and renal artery blood flow in Groups B and C, a decrease in heart rate and mean arterial blood pressure in Group B, and a decrease in aorta blood flow in Group C. CONCLUSIONS: These experimental results demonstrate the usefulness of the comprehensive approach of circulatory changes, and confirm that PEEP may have deleterious effects on regional blood flow, even without significant change in cardiac output, especially when the abdomen is open. PMID- 11861352 TI - Submucous fibrosis secondary to chewing of quids: another cause of unanticipated difficult intubation. AB - PURPOSE: To report a case of unusual difficult intubation secondary to oral submucosal fibrosis (SMF) associated with malignancy of the hypopharynx. CLINICAL FEATURES: A 57-yr-old male, ASA-I physical status presented for percutaneous nephrolithotripsy. The patient was a known alcoholic and smoker with a habit of chewing quids for the past 36 years. Airway assessment revealed oral SMF and slight difficulty in opening the mouth fully. Mentohyoid and mentothyroid distances were normal. However, during intubation, unanticipated difficulty was encountered. On direct laryngoscopy, the tip of the epiglottis could barely be visualized. Intubation was possible after passing a gum elastic bougie blindly under the epiglottis and negotiating a 7.0 internal diameter tracheal tube over it. Postoperatively, indirect laryngoscopy (IDL) showed severe SMF involving the oral cavity, epiglottis and pyriform fossae and an ulcerative malignancy of the hypopharynx with fixation of the left hemilarynx. CONCLUSION: SMF is a premalignant condition of the oral cavity associated with fibrosis of involved structures that can be associated with malignancy of the aerodigestive tract. Distortion of the airway anatomy may render intubation difficult. IDL maybe useful in the preoperative evaluation of these patients. PMID- 11861353 TI - Best evidence in anesthetic practice. Treatment: vasopressin neither improves nor worsens survival from cardiac arrest. PMID- 11861354 TI - Rescue therapy with inhaled nitric oxide in critically ill patients with severe hypoxemic respiratory failure (Brief report). AB - PURPOSE: To evaluate the efficacy of inhaled nitric oxide (iNO) on oxygenation, shunt, and pulmonary vascular resistance index (PVRI) in severely hypoxemic, ventilated patients. METHODS: In a two-period double-blind crossover design, 14 critically ill, hypoxemic, ventilated patients were randomized to receive iNO 10 ppm in 100% oxygen or no iNO in 100% oxygen for 30 min followed by a 30-min washout period and then crossed over to the other intervention. Responders to iNO then received iNO, which was increased from 5 ppm to 25 ppm in 5 ppm increments. Severity of illness scores and cardiorespiratory variables were measured. RESULTS: Nitric oxide decreased shunt (P=0.002) and PVRI (P=0.033) and increased oxygenation (P=0.011) although the latter two were not statistically significant after adjustment for multiple comparisons. Treatment by period interactions were observed. CONCLUSION: Our findings suggest that iNO improves oxygenation to a clinically significant extent in critically ill patients who are severely hypoxemic. PMID- 11861357 TI - Unexpectedly low levels of plasma brain natriuretic peptide in the patient with massive pulmonary tumour emboli. PMID- 11861358 TI - Inadvertent insertion of a gastric tube into the airway in an awake patient. PMID- 11861359 TI - Life threatening external iliac artery injury following total hip replacement. PMID- 11861360 TI - Electroconvulsive therapy with thiamylal or propofol during pregnancy. PMID- 11861361 TI - On smooth extubation without coughing and bucking. PMID- 11861362 TI - Opening propofol ampoules safely. PMID- 11861363 TI - Array-based comparative genomic hybridization for the differential diagnosis of renal cell cancer. AB - Array-based comparative genomic hybridization (CGH) uses multiple genomic clones arrayed on a slide to detect relative copy number of tumor DNA sequences. Application of array CGH to tumor specimens makes genetic diagnosis of cancers possible and may help to differentiate relevant subsets of tumors, biologically and clinically, which would allow better prognostic and therapeutic decision making. In this study, we have used array-based CGH to detect DNA copy number alterations in distinct types of renal cell carcinomas for diagnostic purposes. We were able to correctly diagnose 33 of 34 malignant tumors by automated computational means and to group together eight benign neoplasms and normal kidney samples. These results indicate that array-based CGH is capable of diagnosing the vast majority of renal cell carcinomas based on their genetic profiles. PMID- 11861364 TI - Analysis of gene induction in human fibroblasts and bladder cancer cells exposed to the methylation inhibitor 5-aza-2'-deoxycytidine. AB - Hypermethylation of the promoters of cancer-related genes is often associated with their inactivation during tumorigenesis. Several preclinical and clinical trials have been developed to use DNA methylation inhibitors, such as 5-aza-2' deoxycytidine (5-Aza-CdR) in attempts to reactivate silenced genes in human cancers. We used high-density oligonucleotide gene expression microarrays to examine the effects of 5-Aza-CdR treatment on human fibroblast cells (LD419) and a human bladder tumor cell line (T24). Data obtained 8 days after recovery from 5 Aza-CdR treatment showed that more genes were induced in tumorigenic cells (61 genes induced; >or=4-fold) than nontumorigenic cells (34 genes induced; >or= 4 fold). Approximately 60% of induced genes did not have CpG islands within their 5' regions, suggesting that some genes activated by 5-Aza-CdR may not result from the direct inhibition of promoter methylation. Interestingly, a high percentage of genes activated in both cell types belonged to the IFN signaling pathway, confirming data from other tumor cell types. PMID- 11861365 TI - Genomic amplification in retinoblastoma narrowed to 0.6 megabase on chromosome 6p containing a kinesin-like gene, RBKIN. AB - All retinoblastomas (RBs) show genomic changes in addition to loss of both RB1 alleles. Quantitative-multiplex PCR analysis identified in 41 of 70 (59%) RBs a 0.6-Mb minimal region of chromosome 6p22 gain from which we cloned the human kinesin gene, RBKIN/KIF13A, by rapid amplification of retinal cDNA. RBKIN is expressed ubiquitously in adult tissues, at low levels in fetal tissues, and at high levels in RB. Antisense RBKIN oligonucleotide reduced the growth rate of RB cell lines. RBKIN and/or another closely linked gene are candidate oncogenes contributing to malignant progression of RB. PMID- 11861366 TI - Comprehensive scanning of somatic mitochondrial DNA mutations in breast cancer. AB - To investigate the role of mitochondrial DNA (mtDNA) in tumorigenesis, a temporal temperature gradient gel electrophoresis method was used to analyze the somatic mtDNA mutations in breast cancer. The entire mitochondrial genomes in 19 tumor samples and paired normal tissues from the same patients were amplified using 32 pairs of overlapping primers. DNA fragments showing different banding patterns between normal and tumor mtDNA were sequenced to identify the mutations. Fourteen of the 19 tumors (74%) displayed at least one somatic mtDNA mutation. Twenty seven somatic mutations were found, and 22 of them occurred in the D loop region. This study represents the most comprehensive mtDNA mutational analysis in breast cancer. PMID- 11861367 TI - Enhanced phagemid particle gene transfer in camptothecin-treated carcinoma cells. AB - Engineered phage-based vectors are an attractive alternative strategy for gene delivery because they possess no natural mammalian cell tropism and can be genetically modified for specific applications. Genotoxic treatments that increase the transduction efficiency of single-stranded adeno-associated virus were tested on cells transfected by single-stranded phage. Indeed, green fluorescent protein transgene expression by epidermal growth factor-targeted phagemid particles increased with heat shock, UV irradiation, and camptothecin (CPT) treatment. CPT resulted in transduction efficiencies of 30-45% in certain human carcinoma cell lines and reduced the minimal dose needed to detect green fluorescent protein-expressing cells to as low as 1-10 particles/cell. Targeted phage transduction was effective in many tumor cell lines and in prostate tumor xenografts with CPT treatment. Taken together, these data suggest the feasibility of using phage-based vectors for therapeutic gene delivery to cancer cells. PMID- 11861368 TI - The role of Ral A in epidermal growth factor receptor-regulated cell motility. AB - Tumor cell motility is one of the rate-limiting steps of invasion, which defines progression toward a more malignant phenotype. Elevated expression of epidermal growth factor (EGF) receptor in many cancers is associated with progression of superficial to invasive forms of the disease and is sometimes found in tumors that also have activating Ras mutations, suggesting that both events contribute to tumor invasion. Here we show that EGF stimulates motility in human tumor cell lines, which harbor activating Ha-RasV12 via a novel signal transduction pathway mediated by the small GTP-binding proteins RalA and RhoA but independent of Rac1 and Cdc42. On EGF stimulation, RalA localizes to the cell membrane. In addition, activation of RalA and expression of Rho were increased by EGF stimulation in both the nonmetastatic and metastatic variants of the same cell line. However, elevated levels of constitutively activated RalA were only found in the metastatic variant. This is the first demonstration of an essential role for Ral in EGF-mediated cell motility and its potential contribution to tumor metastasis in human cancer. PMID- 11861369 TI - Trk receptor inhibition induces apoptosis of proliferating but not quiescent human osteoblasts. AB - Prostate cancer frequently metastasizes to the skeleton, producing painful osteoblastic lesions, which are associated with significant morbidity and mortality. This bone tropism involves the bidirectional paracrine interactions between prostate cancer cells and osteoblasts. These interactions enhance prostate cancer cell survival and proliferation of osteoblasts. Therefore, agents that can induce apoptosis of prostate cancer cells and proliferating osteoblasts would be highly advantageous. Previously, we have documented that the unique survival pathway for prostate cancer cells involves a neurotrophin/Trk receptor autocrine pathway. The indocarbazole compounds, CEP-701 and CEP-751, are potent inhibitor of this Trk receptor survival signaling and thus selectively induces apoptosis of prostate cancer cells in various in vitro and in vivo models. In this study, we documented the effects of CEP-751 on the conditionally immortalized osteoblastic cell line, hFOB, in vitro. At the permissive temperature of 34 degrees C, these cells express large T antigen, inducing their continuous proliferation, whereas at 39 degrees C, T antigen is degraded and the cells stop proliferating without undergoing apoptosis. Trk receptors are expressed in hFOB cells, as determined both by reverse transcription-PCR and Western blots. These osteoblasts were shown to produce nerve growth factor and brain-derived neurotrophic factor but not neurotrophin-3, as measured by ELISA. hFOB osteoblasts, cultured at 34 degrees C, secreted significantly (P < 0.01) more brain-derived neurotrophic factor and nerve growth factor into the medium than hFOB cells cultured at 39 degrees C. Because the Trk/neurotrophin axis is present in both proliferating and quiescent (i.e., nonproliferating) osteoblasts, the effects of 48 h of exposure to various doses of CEP-751 on cell viability and apoptosis of hFOB cells were assessed by trypan blue exclusion assays and 4',6 diamidino-2-phenylindole nuclear staining. Cell viability and apoptosis of hFOB cells at 34 degrees C were significantly and dose-dependently decreased compared with untreated proliferating cells. In contrast, even the highest concentration of CEP-751 (200 nM) did not affect cell viability and apoptosis of quiescent hFOB cells cultured at 39 degrees C. This trk inhibition-induced cytotoxicity was confirmed using early-passage, proliferating normal (i.e., non-SV40-transformed) human osteoblasts, which also express Trk receptor protein. These combined results demonstrate that proliferating osteoblasts acquire a sensitivity to trk inhibition- induced apoptosis not shared with normally quiescent osteoblasts. PMID- 11861370 TI - Cancer susceptibility of mice with a homozygous deletion in the COOH-terminal domain of the Brca2 gene. AB - Inherited mutations of the human BRCA2 gene confer increased risks for developing breast, ovarian, and several other cancers. Unlike previously described Brca2 knockout mice that display predominantly embryonic lethal phenotypes, we developed mice with a homozygous germ-line deletion of Brca2 exon 27 that exhibit a moderate decrease in perinatal viability and are fertile. We deleted this Brca2 COOH-terminal domain because it interacts directly with the Rad51 protein, contains a nuclear localization signal, and is required to maintain genomic stability in response to various types of DNA damage. These homozygous Brca2 mutant mice have a significantly increased overall tumor incidence and decreased survival compared with their heterozygous littermates. Virgin female mice homozygous for this Brca2 mutation also display an inhibition of ductal side branching in the mammary gland at 6 months of age. Given their substantial viability and cancer predisposition, these mutant mice will be useful to further define the role of the COOH-terminal Brca2 domain in tumorigenesis both in vivo and in vitro. PMID- 11861371 TI - Recovery of liver mass without proliferation of hepatocytes after partial hepatectomy in Skp2-deficient mice. AB - The abundance of p27(Kip1), an inhibitor of cell proliferation, is determined by Skp2-dependent proteolysis, the deregulation of which is associated with cancer progression. Lack of Skp2 results in p27(Kip1) accumulation as well as enlargement and polyploidy of hepatocytes. The role of Skp2 in cell growth and proliferation was investigated in Skp2-deficient mice subjected to partial hepatectomy. Skp2(-/-) mice exhibited restoration of liver mass without cell proliferation; rather, hepatocytes increased in size, an effect that was accompanied by increased polyploidy and p27(Kip1) accumulation. Lack of Skp2 thus impairs hepatocyte proliferation, which is compensated for by cellular enlargement, during liver regeneration. PMID- 11861372 TI - Increased risk of local recurrence is associated with allelic loss in normal lobules of breast cancer patients. AB - Allelic losses characteristic of tumor cells, when displayed by morphologically normal terminal ductal lobular units (TDLUs) adjacent to carcinoma [G. Deng et al., Science (Wash. DC), 274: 2057-2059, 1996], may indicate an extended field of increased cancer susceptibility within the affected breast tissue. We investigated this possibility by asking whether the presence of loss of heterozygosity (LOH) at chromosome 3p11-26 in histologically normal TDLUs (3pLOHn) could lead to an increased risk of local tumor recurrence. We assessed LOHs in normal TDLUs adjacent to 48 informative cases of early-stage invasive breast cancer samples and found 3pLOHn in approximately 25% (13 of 48) of patients whose tumors had 3pLOH in this region. Our analyses suggest that the most frequent region of LOH is localized at 3p24.3. We also demonstrate, using a Cox proportional hazards regression model, that the presence of 3pLOHn was the only variable significantly related to local tumor recurrence, leading to a 3.9 5.2-fold increase in the hazard ratio (P < 0.05). The time to recurrence was longer in such cases than in those without 3pLOHn, suggesting de novo tumor development. These data provide a strong rationale to assess histologically normal breast tissue at the margins of surgically excised cancers for molecular predictors of local recurrence after breast-conserving treatment. PMID- 11861373 TI - The nonsteroidal anti-inflammatory drug sulindac causes down-regulation of signal transducer and activator of transcription 3 in human oral squamous cell carcinoma cells. AB - The nonsteroidal anti-inflammatory drug sulindac exerts a significant antineoplastic effect on several types of human cancers including oral squamous cell carcinoma (SCCa). Because constitutive activation of signal transducer and activator of transcription 3 (Stat3) has been linked to carcinogenesis of various tumors including head and neck SCCa, we studied whether sulindac treatment affects the Stat3 signaling pathway in oral SCCa cells. Western blot experiments showed that short-term treatment of cells with sulindac resulted in a large reduction of phosphorylated Stat3, without significantly affecting Stat3 protein levels. In contrast, 3 days of sulindac treatment eliminated both phosphorylated and unphosphorylated Stat3 protein levels. Also, sulindac treatment exerted a significant time-dependent cell growth-inhibitory effect on oral SCCa cells under the same conditions shown to induce Stat3 down-modulation. The sulfone metabolite of sulindac, which lacks cyclooxygenase-inhibitory activity, did not affect either Stat3 expression or Stat3 phosphorylation. Antisense oligonucleotide treatment against peroxisome proliferator-activated receptor gamma did not attenuate the ability of sulindac to down-regulate Stat3. Our results suggest that down-modulation of Stat3 can be induced by sulindac treatment, thus possibly contributing to the antineoplastic effect of this drug. PMID- 11861374 TI - Disruption of androgen receptor function inhibits proliferation of androgen refractory prostate cancer cells. AB - Prostate cancer cells depend on androgens and the androgen receptor (AR) for survival. However, after androgen ablation therapy, tumors relapse to an androgen refractory state. To determine whether the androgen receptor is critical for proliferation of androgen-refractory prostate cancer cells, we disrupted the activity of the androgen receptor with an antibody and an AR mRNA hammerhead ribozyme in the following cell lines: LNCaP (androgen-sensitive), LNCaP-Rf and LNCaP-C4 (androgen-refractory), and DU-145 (androgen-insensitive). Microinjection of either antibody or ribozyme inhibited proliferation of androgen-refractory cells. These findings demonstrate that the AR is critical for proliferation of androgen-refractory cells, even in the absence of androgens. PMID- 11861375 TI - Exclusion of breast cancer as an integral tumor of hereditary nonpolyposis colorectal cancer. AB - Hereditary nonpolyposis colorectal cancer (HNPCC) is an autosomal dominant genetic predisposition syndrome that accounts for 2-7% of all colorectal cancers. Diagnosis of HNPCC is based on family history (defined by Amsterdam or Bethesda Criteria), which often includes a history of multiple synchronous or metachronous cancers. The majority of HNPCC results from germ-line mutations in the DNA mismatch repair (MMR) genes hMSH2 and hMLH1 with rare alterations in hMSH6 and hPMS2 in atypical families. Both HNPCC and sporadic MMR-deficient tumors invariably display high microsatellite instability (MSI-H). Two types of HNPCC families can be distinguished: type I (Lynch I) with tumors exclusively located in the colon; and type II (Lynch II) with tumors found in the endometrium, stomach, ovary, and upper urinary tract in addition to the colon. A proposed association of breast cancer with type II HNPCC is controversial. To address this important clinical question, we examined MSI in a series of 27 female patients who presented with synchronous or metachronous breast plus colorectal cancer. Although MSI-H was found in 5 of 27 (18.5%) of the colon cancers, in all cases the matched breast cancer was microsatellite stable. We also examined the breast tumors from three women who were carriers of MMR gene mutations from HNPCC families. None of these three breast tumors displayed MSI nor was the expression of MMR proteins altered in these tumors. We conclude that breast cancer largely arises sporadically in HNPCC patients and is rarely associated with the HNPCC syndrome. PMID- 11861376 TI - Early placenta insulin-like growth factor (pro-EPIL) is overexpressed and secreted by c-erbB-2-positive cells with high invasion potential. AB - By differential-display PCR a subclone of the SKBR3 cell line with high in vitro transendothelial invasiveness was identified to express increased levels of the INSL-4 gene. This new member of the insulin-like growth factor family encodes for a peptide, designated early placenta insulin-like (EPIL), being expressed in the so-called "invasive" phase of the placental development. Immunohistochemistry on tissue microarrays revealed a heterogeneous expression of EPIL in breast cancer tissue and no expression in the surrounding stroma cells. A coexpression of pro EPIL and c-erbB-2 could be observed predominantly in cell clusters at the infiltrating edge of the tumor. Our results give new suggestions for the presence of a signaling network of receptor tyrosine kinases underlying breast cancer invasion and metastasis. PMID- 11861377 TI - Inhibition of activator protein-1 binding activity and phosphatidylinositol 3 kinase pathway by nobiletin, a polymethoxy flavonoid, results in augmentation of tissue inhibitor of metalloproteinases-1 production and suppression of production of matrix metalloproteinases-1 and -9 in human fibrosarcoma HT-1080 cells. AB - Medicinal plants contain pharmacological substances including flavonoids, and their extracts have been therapeutically administered for cancer therapy in vitro and in vivo. We investigated the efficacy of a polymethoxy flavonoid, nobiletin, from Citrus depressa on tumor invasion in vitro. Nobiletin inhibited the tumor invasive activity of human fibrosarcoma HT-1080 cells in the Matrigel model, whereas a similar inhibition was observed upon exogenously adding tissue inhibitors of metalloproteinases (TIMPs)-1 and -2. The gene expression and production of pro-matrix metalloproteinase 9 (proMMP-9)/progelatinase B and proMMP-1/interstitial procollagenase were specifically suppressed by nobiletin in 12-O-tetradecanoylphorbol 13-acetate-stimulated HT-1080 cells. In contrast, the gene expression and production of TIMP-1, but not TIMP-2, were enhanced by nobiletin. We also demonstrated that nobiletin suppressed the 12-O tetradecanoylphorbol 13-acetate-induced binding activity of activator protein-1. Furthermore, a phosphatidylinositol 3-kinase inhibitor, LY-294002, was found to mimic the different actions of nobiletin on the production of proMMP-9 and TIMP 1. These results suggest that nobiletin inhibits tumor cell invasive activity not only by suppressing the expression of MMPs but also augmenting TIMP-1 production in tumor cells, and that the nobiletin-mediated inhibition of activator protein-1 binding activity is at least partly involved in the suppression of MMP expression. Furthermore, we suggest a possible mechanism by which nobiletin may interfere in the phosphatidylinositol 3-kinase pathway, which divergently regulates the production of MMP and TIMP-1. PMID- 11861378 TI - Circulating insulin-like growth factor-I levels regulate colon cancer growth and metastasis. AB - It has been shown previously that slight elevations in serum levels of insulin like growth factor-I (IGF-I) are correlated with an increased risk for developing prostate, breast, colon, and lung cancer. The aim of this study was to determine the role of serum IGF-I levels in the process of stimulating tumor growth and metastasis in a mouse model of colon cancer. Colon 38 adenocarcinoma tissue fragments were orthotopically transplanted by attachment to the surface of the cecum in control and liver-specific IGF-I-deficient (LID) mice in which serum IGF I levels are 25% of that in control mice. A total of 156 male mice at 5 weeks of age (74 control mice and 82 LID mice) received tumor transplants. Mice were divided randomly into two groups; one group was injected i.p. with recombinant human IGF-I (2 mg/kg) twice daily for 6 weeks, and the other group received saline injections. IGF-I treatment increased the serum levels of IGF-I and IGFBP 3 in both control and LID mice. In the saline-injected group, the incidence of tumor growth on the cecum as well as the frequency of hepatic metastasis was significantly higher in control mice as compared with LID mice. Both control and LID mice treated with recombinant human IGF-I displayed significantly increased rates of tumor development on the cecum and metastasis to the liver, as compared with saline-injected mice. The number of metastatic nodules in the liver was significantly higher in control mice as compared with LID mice. The expression of vascular epithelial growth factor (VEGF) as well as vessel abundance in the cecum tumors was dependent on the levels of serum IGF-I. This study supports the hypothesis that circulating IGF-I levels play an important role in tumor development and metastasis. PMID- 11861379 TI - Two forms of gonadotropin-releasing hormone (GnRH) are expressed in human breast tissue and overexpressed in breast cancer: a putative mechanism for the antiproliferative effect of GnRH by down-regulation of acidic ribosomal phosphoproteins P1 and P2. AB - Gonadotropin-releasing hormone (GnRH) analogues are used for the treatment of breast, prostate, and ovarian cancer. These analogues exert their antitumor effects indirectly by inhibiting the pituitary-gonadal axis, as well as by direct inhibition of proliferation of human breast cancer cells. However, the molecular mechanisms mediating these direct antiproliferative effects are not fully understood. We found that normal and malignant human breast tissue express two forms of the neuropeptide GnRH. Quantitative reverse transcription-PCR shows that mRNA encoding the GnRH-I and GnRH-II peptides are overexpressed in cancerous versus normal tissues obtained from the same patients. To elucidate the function of these peptides in breast cancer cells, we used the atlas human cDNA expression arrays technology and studied the differentially regulated genes after GnRH treatment of MCF-7 cells. We found that a wide range of GnRH-I or GnRH-II concentrations (0.1-10 nM) inhibit the expression of mRNA encoding the 60S acidic ribosomal phosphoproteins, P1 and P2. These results were confirmed by quantitative reverse transcription-PCR, as well as Western blot analysis and immunofluorescence staining. The P1 and P2 proteins interact with elongation factors EF1 and EF2, and the level of their phosphorylation is one of the regulatory mechanisms for the overall rate of protein elongation. Thus, reduced expression of P1 and P2 proteins can affect the rate of protein translation, thereby decreasing proliferation rate of cells. Our studies may therefore suggest a putative mechanism for the direct antiproliferative effect of GnRH in breast cancer cells. PMID- 11861380 TI - The androgen receptor CAG repeat polymorphism and risk of breast cancer in the Nurses' Health Study. AB - Shorter alleles of a polymorphic [CAG](n) repeat in exon 1 of the androgen receptor (AR) have been associated with increased risk of prostate cancer and decreased risk of breast cancer. We prospectively assessed the association between the [CAG](n) repeat polymorphism in the androgen receptor and breast cancer risk among Caucasian women in a case-control study nested within the Nurses' Health Study cohort (cases, n = 727; controls, n = 969). In addition, we assessed whether androgen receptor genotype influences endogenous steroid hormone levels in women and whether the associations between androgen receptor alleles and breast cancer risk differed according to established breast cancer risk factors. Women with one or more long AR [CAG](n) repeat alleles (>or=22 repeats) were not at increased risk of breast cancer [odds ratio (OR), 1.06; 95% confidence interval (CI), 0.83-1.35]. Significant associations were not observed between AR genotypes comprised of two short alleles ([CAG](n) or=22: OR, 0.92; 95% CI, 0.62-1.36) or two long alleles ([CAG](n) >or= 25 versus both alleles or=22; OR, 1.70; 95% CI, 1.20-2.40; P for interaction = 0.04). In summary, we observed no overall relation of AR genotype with breast cancer risk among mostly postmenopausal Caucasian women. However, these data suggest that longer AR [CAG](n) repeat alleles may increase breast cancer risk among women with a first-degree family history of breast cancer. PMID- 11861381 TI - Increased vaccination efficiency with apoptotic cells by silica-induced, dendritic-like cells. AB - We have demonstrated previously the ability of apoptotic cells to prime a functional immune response using an i.p. vaccination protocol with apoptotic cells and interleukin 2, before injecting a lethal dose of tumor cells into syngeneic rats. This protocol resulted in a survival rate of 33%. To elucidate the nature and the activity of the phagocytes involved in the clearance of apoptotic cells in vivo, we modulated the peritoneal cavity environment by administrating either thioglycollate or silica i.p. before injecting the apoptotic cells. Our results showed that thioglycollate abrogated vaccination efficiency, because none of the rats survived under these conditions. In fact, thioglycollate treatment induced a massive recruitment and activation of inflammatory macrophages that efficiently engulfed apoptotic cells, bypassing induction of specific immune responses. In contrast, silica treatment enhanced the vaccination efficiency of apoptotic cells plus interleukin 2 up to 66%. We distinguished a population of dendrite-like cells among the cells derived from the silica-treated peritoneal cavity both by their phenotype (MHC II(+)/CD80(+)/CD86(+)) and by their ability to induce the proliferation of allogeneic T cells in a mixed leukocyte reaction. Our results demonstrate the different roles of macrophages and dendritic-like cells in the physiological clearance of dead tumor cells and their implication in the design of immunomodulating vaccines. PMID- 11861383 TI - Targeting of adenovirus vectors to tumor cells does not enable efficient transduction of breast cancer metastases. AB - Targeting of oncolytic adenoviruses to tumors can potentially increase their efficacy and safety profile after systemic application. We have developed recently a capsid-modified vector containing the adenovirus serotype 35 fiber shaft and knob inserted into an Ad5 capsid. This Ad5/35 vector infects cells via a coxsackievirus adenovirus receptor-independent pathway. Here we attempted to exploit this new tropism of Ad5/35 vectors for tumor-specific infection. In vitro, the Ad5/35 vector efficiently transduced human breast cancer cells that were refractory to infection with conventional Ad5-based vectors. Additionally, primary mouse hepatocytes were relatively refractory to Ad5/35 infection in vitro or after systemic vector application to mice. In an animal model of breast cancer metastasis, intraportal infusion of MDA-MB435 cells produced multiple hepatic metastases that were surrounded by extracellular matrix and developed blood vessels confined to the tumor stroma. Tail vein injection of a standard Ad5-based vector into tumor-bearing animals resulted in transduction of mouse hepatocytes but not metastases. However, the capsid-modified Ad5/35 vector transduced only approximately 8% of metastases. The metastases that were susceptible to Ad5/35 infection demonstrated blood vessels in close proximity to tumor nests without extracellular matrix separating endothelial and tumor cells. These findings indicate that transduction of liver metastases not only requires tumor-specific tropism but also new strategies to increase accessibility of tumor cells to systemically applied oncolytic adenoviruses. PMID- 11861382 TI - WK175, a novel antitumor agent, decreases the intracellular nicotinamide adenine dinucleotide concentration and induces the apoptotic cascade in human leukemia cells. AB - We recently developed a class of novel antitumor agents that elicit a potent growth-inhibitory response in many tumor cells cultured in vitro. WK175, a member of this class, was chosen as a model compound that showed strong in vitro efficacy. WK175 interferes with the intracellular steady-state level of NAD(+), resulting in a decreased cellular NAD(+) concentration. We found that WK175 induces apoptotic cell death without any DNA-damaging effect. The apoptotic death signaling pathway initiated by WK175 was examined in detail: mitochondrial membrane potential, cytochrome c release, caspase 3 activation, caspase 3 and poly(ADP-ribose) polymerase cleavage, and the appearance of a sub-G(1) cell cycle population were determined in time course studies in THP-1 (a human monocytic leukemia cell line) cells. We found activation of this cascade after 24 h of treatment with 10 nM WK175. Induction of apoptosis was prevented by bongkrekic acid, Z-Asp-Glu-Val-Asp-fluoromethylketone, and Z-Leu-Glu-His-Asp fluoromethylketone, inhibitors of the mitochondrial permeability transition and of caspase 3 and 9, respectively, but not by Ac-Tyr-Val-Ala-Asp-CHO, a specific caspase 1 inhibitor, suggesting the involvement of the permeability transition pore, caspase 3, and caspase 9 in the WK175-induced apoptotic cascade. These results imply that decreased NAD(+) concentration initiates the apoptotic cascade, resulting in the antitumor effect of WK175. PMID- 11861384 TI - Apoptosis induced by adenovirus-mediated p53 gene transfer in human glioma correlates with site-specific phosphorylation. AB - Therapeutic replacement of the p53 gene using an adenovirus vector (Ad-p53) may be an effective alternative to conventional therapies for the treatment of glioma. We have previously demonstrated that the introduction of Ad-p53 into glioma cells containing mutant p53 induces apoptosis, whereas glioma cells containing wild-type p53 are resistant. However, Ad-p53 will enhance the radiosensitivity of wild-type p53 glioma cells by increasing their tendency for apoptosis. The mechanism underlying these different responses to Ad-p53 has not been elucidated to date. Because phosphorylation of p53 at serines 15, 20, and 392 may play a role in regulating p53-mediated apoptotic activity, we determined the phosphorylation status of exogenous p53 in mutant and wild-type gliomas after Ad-p53 transfer. Monolayer cultures of glioma cell lines expressing mutant p53 (U251 and U373) or wild-type p53 (U87 and D54) were infected with Ad-p53 and analyzed by Western blotting. High levels of exogenous p53 were detected in both cell lines after Ad-p53 transfer. However, only apoptotic mutant p53 cells expressed high levels of phospho-Ser15-p53 and phospho-Ser20-p53. The levels of phospho-Ser15-p53 and phospho-Ser20-p53 were very low in wild-type p53 cells after Ad-p53 infection alone. When wild-type p53 glioma cells were exposed to radiation after Ad-p53 infection, phospho-Ser15-p53 and phospho-Ser20-p53 were detected at high levels, and the cells subsequently underwent apoptosis; no change in serine 392 was detected. The induction of apoptosis and the expression of phospho-Ser15 and phospho-Ser20 in these cells were also enhanced by the combination of Ad-p53 and other DNA-damaging agents such as cisplatin and bichloroethyl nitrosourea. Furthermore, the expression of phospho-Ser15-p53 and phospho-Ser20-p53 correlated with the amount of apoptosis; the apoptotic activity of p53 in glioma cells was partially inhibited by a mutation of p53 at serine 15. These results suggest that phosphorylation of p53 at serine 15 and serine 20 is critical for apoptosis induction in p53 gene therapy for gliomas. PMID- 11861385 TI - The kinase inhibitor PP1 blocks tumorigenesis induced by RET oncogenes. AB - Oncogenic activation of the RET receptor tyrosine kinase is common in different human cancers. We found that the pyrazolo-pyrimidine PP1 inhibited RET-derived oncoproteins with a half maximal inhibitor concentration of 80 nM. Furthermore, RET/PTC3-transformed cells treated with 5 microM of PP1 lost proliferative autonomy and showed morphological reversion. PP1 prevented the growth of two human papillary thyroid carcinoma cell lines that carry spontaneous RET/PTC1 rearrangements and blocked anchorage-independent growth and tumorigenicity in nude mice of NIH3T3 fibroblasts expressing the RET/PTC3 oncogene. These findings suggest targeting RET oncogenes with PP1 or related compounds as a novel treatment strategy for RET-associated neoplasms. PMID- 11861386 TI - Proteasome inhibitor PS-341, a potential therapeutic agent for adult T-cell leukemia. AB - Nuclear factor kappaB (NF-kappaB) plays a major role in the pathogenesis of human T-cell lymphotrophic virus I-associated malignancy. Proteasome inhibitors provide a rational approach to control constitutively activated NF-kappaB in human T-cell lymphotrophic virus I-infected T cells. We report that the proteasome inhibitor PS-341 decreased NF-kappaB DNA binding activity by preventing degradation of IkappaB(alpha). In our murine model of adult T-cell leukemia, PS-341 used alone did not yield prolongation of the survival of tumor-bearing mice. However, when combined with the current clinically approved drug humanized anti-Tac, therapy with PS-341 was associated with a complete remission in a proportion of treated animals, whereas only a partial response was observed in animals treated with humanized anti-Tac alone. PMID- 11861387 TI - Inhibition of phosphatidylinositol 3'-kinase increases efficacy of paclitaxel in in vitro and in vivo ovarian cancer models. AB - Phosphatidylinositol 3'-kinase (PI3k) is implicated in a wide array of biological and pathophysiological responses. Thus, inhibiting molecules involved in its signal transduction pathway is a possible means of treating cancer. Our previous studies demonstrated that LY294002, a potent and selective PI3k inhibitor, decreases growth of ovarian carcinoma and ascites formation in an athymic mouse xenogeneic transplant model of ovarian cancer. However, the dose of LY294002 used to decrease tumor growth resulted in significant dermatological toxicity. We demonstrate herein that introduction of an active catalytic subunit of PI3k into an ovarian cancer cell line, and thus activation of the PI3k/AKT pathway, confers resistance to the effects of paclitaxel, one of the major drugs used in ovarian cancer therapy. The resistance to paclitaxel can be reversed in vitro by inhibition of PI3k. Therefore, we evaluated whether combined therapy with paclitaxel and LY294002 would result in increased efficacy and allow utilization of doses of LY294002 that do not induce dermatological toxicity. Two weeks after i.p. inoculation with OVCAR-3 ovarian cancer cells, mice were treated i.p. with LY294002 plus paclitaxel, each three times weekly on alternate days, for 4 weeks. Tumor burden in the LY294002 + paclitaxel, LY 294002 alone, and paclitaxel alone groups was reduced by 80% (P < 0.01), 38% (P < 0.05), and 51% (P < 0.05), respectively, compared with controls. Virtually no ascites developed in the combined treatment group; mean volume of ascites in the controls was 3.7 ml. Treatment with LY294002 alone reduced ascites by 70% (P < 0.01), whereas paclitaxel alone reduced ascites slightly but not significantly. No dermatological lesions or weight loss were observed in any treatment group. In vivo and in vitro morphological studies demonstrated that inhibition of PI3k enhanced paclitaxel-induced apoptosis in the human ovarian cancers. Our data suggest that a combination of a PI3k inhibitor and conventional chemotherapy may provide an effective approach to inhibiting tumor growth and ascites production in ovarian cancer with acceptable side effects. PMID- 11861388 TI - The CC chemokine RANTES in breast carcinoma progression: regulation of expression and potential mechanisms of promalignant activity. AB - Breast cancer progression may be affected by various cellular components expressed by the tumor cells and/or by microenvironmental factors. Many studies report the correlation between breast cancer progression and monocyte infiltration into the tumor site. We have identified recently the CC chemokine regulated on activation, normal T cell expressed and secreted (RANTES), a major monocyte chemoattractant expressed by breast tumor cells, as a potential contributor to breast cancer progression. In the present study, analysis of the regulation of RANTES expression demonstrates that the expression of RANTES in breast tumor cells is elevated significantly and in a synergistic manner by IFN gamma and tumor necrosis factor-alpha. Identification of the mechanisms by which RANTES may contribute to breast cancer progression included the analysis of the potential ability of RANTES to act in paracrine and indirect mechanisms, as well as directly on the tumor cells, to promote disease progression. Our results suggest that breast tumor cell-derived RANTES may promote breast cancer progression by its partial contribution to monocyte migration into breast tumor sites. Moreover, RANTES promotes the expression of matrix metalloproteinase (MMP) 9 by THP-1 monocytic cells and elevates vascularity in chick chorioallantoic membrane assays. Tumor necrosis factor-alpha, a major monocyte-derived cytokine, was found to promote the expression of MMP9 and MMP2 by MCF-7 and T47D breast adenocarcinoma cells, respectively, and to induce the de novo expression of an additional proteolytic enzyme by T47D cells, presumably MMP9. The possibility that RANTES may act directly on breast tumor cells was supported by detection of the expression of the CCR5 RANTES receptor in biopsy sections of breast cancer patients and by the ability of RANTES to promote the expression of MMP9 by MCF-7 cells. In all, our study suggests that the expression of RANTES by breast tumor cells results not only in monocyte migration to the tumor site but also in protumorigenic activities of RANTES and of proinflammatory cytokines that may facilitate metastasis formation and contribute to disease progression. PMID- 11861389 TI - IL-13R(alpha)2, a decoy receptor for IL-13 acts as an inhibitor of IL-4-dependent signal transduction in glioblastoma cells. AB - Interleukin (IL)-4 and IL-13 share the type II IL-4 receptor for cell signaling. We show that despite expressing the necessary signaling components, glioblastoma cells failed to respond to either IL-4 or IL-13. This was in part because of the expression of a high-affinity IL-13-binding transmembrane protein IL-13R(alpha)2 that inhibited IL-13-mediated Stat6 activation by acting as a decoy receptor. In contrast, normal human astrocytes that did not express the IL-13R(alpha)2 gene efficiently induced Stat6 activation in response to both IL-4 and IL-13. Transient expression of the IL-13R(alpha)2 transgene in nonexpressing heterologous cells inhibited not only IL-13- but also IL-4-mediated signal transduction and Stat6-responsive gene expression. The inhibition was likely mediated through the physical interaction between the short intracellular domain of the IL-13R(alpha)2 protein and the cytoplasmic domain of the IL-4R(alpha) chain that harbors the Stat6 docking sites. Thus, IL-13R(alpha)2 acts as an inhibitor of IL-4-dependent signal transduction pathways via a novel mechanism that is independent of ligand binding. PMID- 11861391 TI - Apoptosis-dependent subversion of the T-lymphocyte epitope hierarchy in lymphoma cells. AB - Tumor cells undergoing programmed death are an attractive source of tumor associated antigens, and evidences are available for their therapeutic efficacy in vivo when used either alone or in association with dendritic cells. However, little is known about the specificity of the immune response induced by such antigen formulation. Indeed, activation of specific proteases during apoptosis may influence the cytoplasmic degradation of proteins and the generation of CTL epitopes. We show here that on injection of C57BL/6 mice either with RMA lymphoma cells induced to apoptosis or bone marrow-derived dendritic cells pulsed with apoptotic RMA cells, a specific and protective CTL response is induced, which, however, is not directed against the immunodominant CTL epitope gag(85-93). Lack of in vivo expansion of gag(85-93)-specific CTL in vaccinated mice is attributable to the apoptosis-dependent loss of gag(85-93) in dying tumor cells. Indeed, we found loss of gag(85-93) in RMA, MBL-2, and EL-4G+ lymphoma cells, which share gag(85-93) as an immunodominant CTL epitope, induced to apoptosis by UV irradiation, mitomycin C, doxorubicin, or daunorubicin. This phenomenon appears to be caspase-dependent, because caspase inhibition by N benzyloxycarbonyl-Val-Ala-asp-fluoromethylketone prevents apoptosis of lymphoma cells and loss of gag(85-93). Therefore, subversion of the epitope hierarchy in apoptotic tumor cells might be relevant in the induction of tumor-specific T lymphocyte responses. PMID- 11861390 TI - A tumor-expressed inhibitor of the early but not late complement lytic pathway enhances tumor growth in a rat model of human breast cancer. AB - Membrane-bound complement inhibitors protect host cells from inadvertent complement attack, and complement inhibitors are often up-regulated on tumors, possibly representing a selective adaptation by tumors to escape elimination by a host antitumor immune response. Relevant in vivo studies using rodent models of human cancer have been hampered by the fact that human complement inhibitors are not effective against rodent complement. Using nude rats and MCF7 cells expressing different rat complement inhibitors, a model of human breast cancer was established to investigate the role of complement and complement inhibitors in tumor progression. Expression of rat CD59, an inhibitor of the terminal cytolytic membrane attack complex of complement, had no effect on the incidence or growth rate of MCF7 tumors. In contrast, expression of rat Crry, an inhibitor of complement activation, dramatically enhanced the tumorigenicity of MCF7 cells. The expression of rat Crry on MCF7 inhibited the in vivo deposition of complement C3 fragments that serve as opsonins for receptors on phagocytes and natural killer cells. These data provide direct in vivo evidence that an inhibitor of complement activation can facilitate tumor growth by modulating C3 deposition. These data indicate an important role for complement opsonization in promoting cell-mediated antitumor immune function, a conclusion further supported by the demonstration that expression of rat Crry, but not rat CD59, on MCF7 cells inhibits rat cell-mediated cytotoxicity in vitro. Rat complement activation on MCF7 tumors was mediated by tumor-reactive antibodies present in the serum of naive nude rats, but there was also an IgM response to MCF7 tumors, a situation with similarities to some human cancers. These data support a hypothesis that blocking complement inhibitor function on tumor cells will not only enhance monoclonal antibody-mediated immunotherapy but may also be effective at enhancing a normally ineffective humoral immune response in the absence of administered antitumor antibody. PMID- 11861392 TI - Minichromosome maintenance protein MCM7 is a direct target of the MYCN transcription factor in neuroblastoma. AB - The MYCN oncogene is amplified in approximately 25% of neuroblastoma tumors and is the most significant negative prognostic factor. The direct transcriptional targets of MYCN in MYCN-amplified tumors have not been defined. Microarray analysis of RNA from neuroblastoma primary cell cultures revealed 10-fold higher MCM7 expression in MYCN-amplified versus nonamplified tumors. MCM7 is an essential component of DNA replication licensing factor, a hexameric protein complex that regulates DNA synthesis during the cell cycle, preventing rereplication and ensuring maintenance of DNA euploidy. Additional experiments demonstrated markedly increased expression of MCM7 RNA and protein in MYCN amplified neuroblastoma tumors and cell lines. Induction of MYCN in conditional cell lines results in increased expression of endogenous MCM7 mRNA and a 3-fold increase in protein levels. In addition, luciferase activity from MCM7 promoter/luciferase gene reporter constructs was significantly increased under MYCN-induced conditions. Specific electrophoretic mobility shifts of MCM7 promoter sequences are detected in extracts of MYCN-amplified cells. These findings demonstrate that in neuroblastoma, the MYCN oncogene directly activates genes required for DNA replication, and this may contribute to neoplastic transformation of these MYCN-amplified tumors. PMID- 11861393 TI - Targeted inactivation of p53 in human cells does not result in aneuploidy. AB - Because p53 mutation and aneuploidy usually coexist, it has been suggested that p53 inactivation leads to aneuploidy. We have rigorously tested this hypothesis in diploid human cell lines in which p53 was experimentally inactivated by targeted homologous recombination. Cells completely deficient in p53 did not become aneuploid, although a slight tendency toward tetraploidization was observed. No increased rates of numerical or structural chromosomal instabilities were observed in the p53-deficient cells. Rates of sister chromatid exchange and homologous recombination were also unaffected by p53 status. These results show that inactivation of p53 does not, in and of itself, lead to the development of aneuploidy. PMID- 11861394 TI - Silence of chromosomal amplifications in colon cancer. AB - Oncogene activation by gene amplification is a major pathogenetic mechanism in human cancer. Using comparative genomic hybridization, we determined that metastatic human colon cancers commonly acquire numerous extra copies of chromosome arms 7p, 8q, 13q, and 20q. We then examined the consequence of these amplifications on gene expression using DNA microarrays. Of 55,000 transcripts profiled, 2,146 were determined to map to one of the four common colon cancer amplicons and to also be expressed in normal or malignant colon tissues. Of these, only 81 transcripts (3.8%) demonstrated a 2-fold increase over normal expression among cancers bearing the corresponding chromosomal amplification. Chromosomal amplifications are common in colon cancer metastasis, but increased expression of genes within these amplicons is rare. PMID- 11861395 TI - An integrated database of chemosensitivity to 55 anticancer drugs and gene expression profiles of 39 human cancer cell lines. AB - To explore genes that determine the sensitivity of cancer cells to anticancer drugs, we investigated using cDNA microarrays the expression of 9216 genes in 39 human cancer cell lines pharmacologically characterized on treatment with various anticancer drugs. A bioinformatical approach was then exploited to identify genes related to anticancer drug sensitivity. An integrated database of gene expression and drug sensitivity profiles was constructed and used to identify genes with expression patterns that showed significant correlation to patterns of drug responsiveness. As a result, sets of genes were extracted for each of the 55 anticancer drugs examined. Whereas some genes commonly correlated with various classes of anticancer drugs, other genes correlated only with specific drugs with similar mechanisms of action. This latter group of genes may reflect the efficacy of each class of drugs. Therefore, the integrated database approach of gene expression and chemosensitivity profiles may be useful in the development of systems to predict drug efficacies of cancer cells by examining the expression levels of particular genes. PMID- 11861396 TI - Hypermethylation of the p14(ARF) gene in ulcerative colitis-associated colorectal carcinogenesis. AB - The p14(ARF) protein directly inhibits the MDM-2 oncoprotein, which mediates degradation of the p53 protein. It has been shown that p14(ARF) expression is frequently down-regulated by p14(ARF) gene hypermethylation in colorectal cancer. To determine whether p14(ARF) inactivation was involved in ulcerative colitis (UC)-associated carcinogenesis, the frequency and timing of p14(ARF) methylation was investigated in four different histological stages of UC-associated carcinogenesis. Methylation-specific PCR and bisulfite sequencing were used to determine the prevalence of p14(ARF) gene methylation. p14(ARF) methylation was observed in 19 of 38 (50%) adenocarcinomas, 4 of 12 (33%) dysplasias, and 3 of the 5 (60%) nonneoplastic UC mucosae. In contrast, 3 of 40 (3.7%) normal tissues showed p14(ARF) methylation (chi(2) test: P = 0.0003). Bisulfite sequencing was used to analyze 28 CpGs of p14(ARF) gene in 20 samples. The number of methylated CpGs ranged from 0 to 4, 0 to 20, and 0 to 28 in the normal, dysplastic, and carcinomatous samples, respectively (Kruskall-Wallis test: P = 0.0005). Densely methylated alleles were detected only in carcinomas by bisulfite sequencing. In conclusion, our data suggest that methylation of p14(ARF) is a relatively common early event in UC-associated carcinogenesis. p14(ARF) offers potential as a biomarker for the early detection of cancer or dysplasia in UC. Finally, analyses of p14(ARF) methylation in other organs should explore not only frank cancers but other premalignant lesions. PMID- 11861397 TI - Specific chromosomal aberrations in mouse lung adenocarcinoma cell lines detected by spectral karyotyping: a comparison with human lung adenocarcinoma. AB - Although adenocarcinoma is rapidly becoming the most common form of lung cancer in the United States, the difficulty in obtaining lung cancer families and representative samples of the various stages of adenocarcinoma progression has led to intense study of mouse models. As a powerful approach to delineating molecular changes, we have analyzed 15 early-passage mouse cell lines by spectral karyotyping. Entire copies of chromosomes 1, 2, 6, 12, 15, and 19 were gained, and entire copies of chromosomes 4, 7, 8, and 14 were lost. Significant gains of portions of chromosome 1 (93% of the tumor cell lines analyzed), chromosome 2 (53%), chromosome 6 (73%), chromosome 7 (80%), chromosome 12 (47%), and chromosome 15 (73%) and partial loss of chromosome 4 (87%), chromosome 7 (80%), chromosome 8 (53%), chromosome 10 (33%), and chromosome 14 (33%) were observed. Recurrent translocations included 10:del(10)(A1::C1), t(4;8)(C4;A1), and der (1;12)(C2;C2). The minimal regions of chromosomal alteration, 1G1, 2F1, 4C4, 6D, 7F1, 8B3, and 12C2, contain putative susceptibility genes for mouse lung adenocarcinoma. Chromosomal regions containing susceptibility genes linked to tumor size were frequently amplified, whereas regions with susceptibility loci linked to tumor multiplicity were deleted. Similar linkage groups are altered in human lung adenocarcinoma, implying that the mouse is a valid genetic model for the study of human lung adenocarcinoma susceptibility. PMID- 11861398 TI - Repression of alpha-fetoprotein gene expression under hypoxic conditions in human hepatoma cells: characterization of a negative hypoxia response element that mediates opposite effects of hypoxia inducible factor-1 and c-Myc. AB - Hypoxia is an important component of many pathological processes including cancerogenesis and cirrhosis. We have attempted to identify additional hepatic genes sensitive to hypoxia by postulating that genes with possible binding sites for hypoxia inducible factor-1 (HIF-1) are regulated by hypoxia. A computer analysis identified the oncodevelopmental alpha-fetoprotein gene (afp) as one of them. The amounts of both alpha-fetoprotein mRNA and protein were decreased under hypoxic conditions in HepG2 hepatoma cells. Stability of afp mRNA was not altered, and de novo synthesis of proteins was required. Transfection experiments in HepG2 cells showed that both hypoxia and overproduction of HIF-1alpha specifically repressed the transcriptional activity of the rat afp regulatory region through the sequence 5'-CACGTGGG-3' located at -3625 to -3619. Mutation in this sequence strongly impaired these repressions. Interestingly, this sequence was a functional stimulatory target for c-Myc, suggesting that c-Myc regulates afp gene expression. Lastly, the amounts of c-myc mRNA and protein were reduced when these cells were grown under hypoxic conditions. Taken together, these results suggest the existence of a possible competition between HIF-1 and c-Myc that could modulate the transcriptional activity of the afp gene in response to hypoxia. PMID- 11861399 TI - Low-level microsatellite instability in most colorectal carcinomas. AB - Twelve to 16% of colorectal cancers (CRCs) display a high degree of microsatellite instability (MSI-H), whereas most are believed to be microsatellite stable (MSS). The existence of a low degree of instability (MSI-L) group has also been proposed. By using the Bethesda panel of microsatellite markers, the microsatellite instability (MSI) status of CRCs can be determined. This set is recommended to distinguish between MSI-H and MSI-L/MSS. No definition for MSI-L has emerged. Most reports on MSI-L rely on the Bethesda panel, using 5 15markers. Tumors with more than 30% MSI are designated as MSI-H, but the lower limit for MSI-L is ambiguous. We hypothesized that if many markers are studied, almost all CRCs would show some MSI. It would be necessary to establish a cutoff level for MSI-L by showing that, above this cutoff level, tumors display molecular and/or clinical features different from those under the cutoff level. To perform this task, we analyzed 90 BAT26 stable CRC samples with 377 markers. MSI at 1-11 loci was observed in 71 (79%) of the 90 cases. K-RAS mutation, loss of heterozygosity, and MLH1 and MGMT hypermethylation analyses were performed, as well as clinical features being scrutinized, to examine possible differences between MSI-L and MSS tumors using all of the possible cutoff levels for MSI-L. Convincing differences between putative MSI-L and MSS groups were not observed. Our results show that the sensitivity of a typically used marker number to detect MSI-L is very low, and they suggest that MSS and MSI-L tumors have a common molecular background. PMID- 11861400 TI - Human tumor suppressor ARF impedes S-phase progression independent of p53. AB - Using alternative reading frames, the human ARF-INK4a locus encodes two unrelated proteins that both function in tumor suppression. p16(INK4a) maintains the retinoblastoma protein in its growth-suppressive state through inhibition of cyclin D-dependent kinase activity, whereas ARF binds with MDM2 and stabilizes p53. The majority of the activity of ARF to date is ascribed to its ability to activate p53, resulting in a G(1) cell cycle arrest or apoptosis. We show here that ARF colocalizes with DNA replication protein A (RPA32) and that overexpression of ARF reduces the rate of DNA synthesis resulting in accumulation of an S-phase cell population. Impediment of DNA synthesis by ARF can occur and becomes more evident in the absence of p53. Hence, the biological consequence of ARF induction varies dependent on cellular p53 status, inducing predominantly a G(1) arrest or apoptosis in p53-positive cells or causing S-phase retardation when p53 function is comprised. PMID- 11861401 TI - GATA-6 transcriptional regulation of 15-lipoxygenase-1 during NSAID-induced apoptosis in colorectal cancer cells. AB - The mechanisms by which nonsteroidal anti-inflammatory drugs (NSAIDs) induce apoptosis in colorectal cancer cells are undergoing intensive investigation. We found previously that NSAIDs induce apoptosis in these cells by restoring 15 lipoxygenase-1 (15-LOX-1) expression. The present study examined the NSAID mechanism for up-regulating 15-LOX-1 in two colorectal cancer cell lines (RKO and DLD-1). We found that NSAID effects on 15-LOX-1 occurred at the level of transcriptional regulation. We then studied NSAID effects on GATA-6, a transcription factor that suppresses 15-LOX-1 expression. Beginning within 4 h, NSAIDs progressively down-regulated GATA-6 expression. Ectopic GATA-6 expression blocked NSAID induction of 15-LOX-1 and apoptosis. NSAIDs down-regulate GATA-6 to transcriptionally up-regulate 15-LOX-1 and induce apoptosis in colorectal cancer cells. PMID- 11861402 TI - Molecular classification of head and neck squamous cell carcinoma using cDNA microarrays. AB - Squamous cell carcinomas of the head and neck constitute an anatomically heterogeneous group of neoplasms that share in common a causal association with tobacco and alcohol exposure. The clinical course of these neoplasms is difficult to predict based on established prognostic clinicopathological criteria. Given the genetic complexity of head and neck cancers, it is not surprising that correlations with individual genetic abnormalities have also been disappointing. Several authors have suggested that global gene expression patterns can be used to subgroup patients with cancer. Here we report the use of cDNA microarrays containing 9216 clones to measure global patterns of gene expression in these neoplasms. We have used a statistical algorithm to identify 375 genes, which divide patients with head and neck tumors into two clinically distinct subgroups based on gene expression patterns. Our results demonstrate that gene expression profiling can be used as a predictor of outcome. PMID- 11861403 TI - Overexpression of thrombospondin-1 reduces growth and vascular index but not perfusion in glioblastoma. AB - Little is known about the effects of antiangiogenic therapy on perfusion of human tumors and the mechanisms by which tumors can adapt to these treatments and recur. Here, we examined the effects of serial passaging of LN-229 human glioma xenografts overexpressing thrombospondin (TSP)-1 on tumor growth, vascularity, and perfusion. Persistence of TSP-1 overexpression was confirmed after three serial s.c. passages of small xenografted tumor blocks of cells stably transfected with TSP-1 cDNA (clones C9 and E7) or vector controls (pooled clones A7-A9) in immunodeficient nu/nu mice. The tumor vascularity was estimated by noninvasive near infrared spectroscopy measuring blood volume at 800 +/- 10 nm and by histological vessel scores in CD31-immunostained cryosections. The tumor perfusion was assessed by noninvasive laser Doppler flowmetry. Overexpression of TSP-1 significantly inhibited tumor growth. In size-matched tumors (approximately 300 mm(3)), the blood volume and the histological vessel scores were lower in the TSP-1-transfected tumors than in controls, and this effect was more pronounced in tumors derived from the clone with the highest TSP-1 expression (clone E9). Despite this clear reduction in tumor vascularity, the tumor perfusion was the same in TSP-1-transfected tumors and controls. This study shows that TSP-1 overexpression slows glioma growth in vivo but does not prevent it from reaching a large size (300 mm(3)). Whereas a clear reduction in blood volume during tumor growth and a reduced vascular index at sacrifice are observed in TSP-1 transfected tumors, this did not affect perfusion when size-matched comparisons were performed. Given the increased time needed to reach equal size, it indicates that a fixed rate of perfusion must be maintained in the tumor to allow for growth. Elucidation of the mechanisms that allow this to happen has important consequences for the understanding of tumor recurrence after antiangiogenic therapy. PMID- 11861404 TI - retSDR1, a short-chain retinol dehydrogenase/reductase, is retinoic acid inducible and frequently deleted in human neuroblastoma cell lines. AB - Vitamin A is required for a number of developmental processes and for the homeostatic maintenance of several adult differentiated tissues and organs. In human neuroblastoma (NB) cells as well as some other tumor types, pharmacological doses of retinoids are able to control growth and induce differentiation in vitro and in vivo. In a search for new genes that are regulated by retinoids and that contribute to the biological effects retinoids have on NB cells, we have isolated five differentially expressed transcripts. Here we report on the characterization of one of them (DD83.1) in NB cell lines. DD83.1 is identical to the human retSDR1, a short chain dehydrogenase/reductase that is thought to regenerate retinol from retinal in the visual cycle. Its expression is strongly, but differently, regulated by retinoids in NB cell lines, and it is widely expressed in human tissues, which suggests that it is involved in a more general retinol metabolic pathway. Both the retinoic acid-dependent and the exogenous expression of retSDR1 in SK-N-AS cells induce the accumulation of retinyl esters, which indicates that it is involved in generating storage forms of retinol in tissues exposed to physiological retinol concentrations. We also show that the human retSDR1 gene, which maps on chromosome 1p36.1, is contained in the DNA fragment deleted in many NB cell lines bearing MYCN amplification but is conserved in a cell line with a small 1p deletion and normal MYCN. Our observations suggest that retSDR1 is a novel regulator of vitamin A metabolism and that its frequent deletion in NB cells bearing MYCN amplification could compromise the sensitivity of those cells to retinol, thereby contributing to cancer development and progression. PMID- 11861405 TI - Overexpression of copper zinc superoxide dismutase suppresses human glioma cell growth. AB - Copper zinc superoxide dismutase (CuZnSOD) is an essential primary antioxidant enzyme that converts superoxide radical to hydrogen peroxide and molecular oxygen in the cytoplasm. Cytosolic glutathione peroxidase (GPx) converts hydrogen peroxide into water. The overall goal of the present study was to explore the possible role of the antioxidant enzyme CuZnSOD in expression of the malignant phenotype. We hypothesized that overexpression of CuZnSOD would lead to the suppression of at least part of the human malignant phenotype. To test this hypothesis, human CuZnSOD cDNA was transfected into U118-9 human malignant glioma cells. CuZnSOD activity levels increased 1.5-, 2.0-, 2.6-, and 3.5-fold, respectively, in four table transfected cell lines compared with wild type and vector controls. Overexpression of CuZnSOD altered cellular antioxidant enzyme profiles, including those of manganese superoxide dismutase, catalase, and GPx. The transfected clone with the highest CuZnSOD:GPx ratio (3.5) showed a 42% inhibition of tumor cell growth in vitro. The decreased rate of tumor cell growth in vitro was strongly correlated with the enzyme activity ratio of CuZnSOD:GPx. Glioma cells that stably overexpressed CuZnSOD demonstrated additional suppressive effects on the malignant phenotype when compared with the parental cells and vector controls. These cells showed decreased plating efficiency, elongated cell population doubling time, lower clonogenic fraction in soft agar, and, more significantly, inhibition of tumor formation in nude mice. This work suggested that CuZnSOD is a new tumor suppressor gene. Increased intracellular ROS levels were found in cells with high activity ratios of CuZnSOD:GPx. Change in the cellular redox status, especially change attributable to the accumulation of hydrogen peroxide or other hydroperoxides, is a possible reason to explain the suppression of tumor growth observed in CuZnSOD-overexpressing cells. PMID- 11861406 TI - Activation of nuclear factor kappaB in radioresistance of TP53-inactive human keratinocytes. AB - Many tumors show a mutant or inactive tumor suppressor p53 (TP53) status, and TP53 in the tumor-carrying human papillomavirus (HPV) may be dysfunctional because of inhibition by the viral protein HPV E6. Molecular mechanisms underlying radiation responses and the radiation-induced resistant phenotype in the TP53-inactive tumor have not been well investigated. In the present study, using a human keratinocyte line (HK18) with TP53 inhibited by HPV18 infection, we demonstrated that nuclear factor (NF)-kappaB is responsible for a major portion of the radioresistance observed in a cell population (HK18-IR) derived from HK18 cells by fractionated ionizing radiation (FIR; 2 Gy/fraction; total dose, 60 Gy). HK18-IR cells showed increased clonogenic radioresistance [dose-modifying factor (DMF), 1.47], reduced apoptotic response, and a shortened radiation-induced growth delay. Both DNA-binding and reporter transcriptional activity of NF kappaB, but not of TP53, were activated in HK18-IR cells compared with the parental HK18 cells; this activation was observed both before and after a single dose of 5 Gy. To determine target genes responsive to NF-kappaB activation, DNA microarray profiles for 588 genes were matched in HK18-IR cells compared with those in HK18 cells; the paired comparisons were made for basal levels before irradiation or for levels 24 h after 5 Gy. For 25 genes, a 2- to 5-fold up regulation in HK18-IR cells relative to HK18 cells was similar when comparisons were made for basal levels or for levels after irradiation. Included in the approximately 4% of genes activated in HK18-IR cells, were six genes (Cyclin B1, Cyclin D1, HIAP, BAG-1, TTF, and fibronectin) putatively linked to NF-kappaB regulation. We then measured the expression of this group of FIR-regulated genes in HK18-IR cells expressing a dominant-negative mutant IkappaB (mIkappaB) that inhibited NF-kappaB activation. Clonogenic radioresistance was reduced greatly in the mIkappaB transfectants (DMF, 1.18 and 1.10, respectively, at 10% and 1% of isosurvival for mIkappaB transfectants compared with 1.47 and 1.45, respectively, for vector control transfectants). Expressions of Cyclin B1, Cyclin D1, and HIAP were down-regulated by the inhibition of NF-kappaB. These results suggest that transcription of NF-kappaB and a group of NF-kappaB target genes are involved in radioresistance in FIR-treated tumor cells with inactive TP53. PMID- 11861407 TI - The RING domain of Mdm2 can inhibit cell proliferation. AB - Mdm2 is a p53-inducible phosphoprotein that negatively regulates p53 by binding to it and promoting its ubiquitin-mediated degradation. Alternatively spliced variants of Mdm2 have been isolated from human and mouse tumors, but their roles in tumorigenesis, if any, remain elusive. We cloned six alternatively spliced variants of Mdm2 from E(mu)-Myc-induced mouse lymphomas, all of which lacked the NH(2)-terminal p53-binding domain but conserved the remainder of the Mdm2 protein. Enforced expression of full-length Mdm2 in primary mouse embryo fibroblasts or bone marrow-derived, interleukin 7-dependent pre-B cells accelerated their proliferation, whereas unexpectedly, overexpression of truncated Mdm2 isoforms inhibited their growth. Truncated variants were active as inhibitors whether they localized predominantly to the nucleus or cytoplasm. Despite the absence of the p53-binding domain, growth inhibition remained strictly p53 dependent (but not p19(Arf) dependent) and could be overcome by full length Mdm2. The intact RING finger domain at the Mdm2 COOH terminus (amino acids 399-489) was necessary and sufficient for growth inhibition by truncated Mdm2 proteins and could physically interact with either the RING finger domain or central acidic region of full-length Mdm2. However, such interactions do not inhibit Mdm2 E3 ubiquitin ligase activity in vitro using p53 as a substrate. Expression of growth-inhibitory Mdm2 isoforms in tumors remains an enigma. PMID- 11861408 TI - Adipose rex: fat and fats that rule differentiation. PMID- 11861409 TI - MIghty mouse. PMID- 11861410 TI - Beta-myosin heavy chain gene mutations in familial hypertrophic cardiomyopathy: the usual suspect? PMID- 11861411 TI - Transient receptor potential channels regulate myogenic tone of resistance arteries. AB - Elevation of intravascular pressure causes depolarization and constriction (myogenic tone) of small arteries and arterioles, and this response is a key element in blood flow regulation. However, the nature of pressure-induced depolarization has remained elusive. In the present study, we provide evidence that a transient receptor potential channel (TRPC6) homologue has a major role in this depolarizing response to pressure. Antisense oligodeoxynucleotides to TRPC6 decreased TRPC6 protein expression and greatly attenuated arterial smooth muscle depolarization and constriction caused by elevated pressure in intact cerebral arteries. Suppressing the expression of this channel protein also reduced the current density of a major cation current in resistance artery smooth muscle cells. We propose that TRPC6 channels play an essential role in regulation of myogenic tone. PMID- 11861412 TI - Matrix metalloproteinases in vascular remodeling and atherogenesis: the good, the bad, and the ugly. AB - Vascular remodeling, defined as any enduring change in the size and/or composition of an adult blood vessel, allows adaptation and repair. On the other hand, inappropriate remodeling, including its absence, underlies the pathogenesis of major cardiovascular diseases, such as atherosclerosis and restenosis. Since degradation of the extracellular matrix scaffold enables reshaping of tissue, participation of specialized enzymes called matrix metalloproteinases (MMPs) has become the object of intense recent interest in relation to physiological ("good") and pathological ("bad") vascular remodeling. Experimental evidence acquired in vitro and in vivo suggests that the major drivers of vascular remodeling, hemodynamics, injury, inflammation, and oxidative stress, regulate MMP expression and activity. Alternatively, nonspecific MMP inhibition seems to oppose remodeling, as suggested by the inhibition of intimal thickening and outward arterial remodeling. An emerging concept is that MMP-related genetic variations may contribute to heterogeneity in the presentation and natural history of atherosclerosis. The hypothesis that MMPs contribute to weakening of atherosclerotic plaques is especially attractive for the potential development of therapeutic interventions aimed at preventing plaque disruption ("the ugly"), a major cause of acute cardiovascular events. However, the current lack of appropriate experimental tools, including availability of specific MMP inhibitors and pertinent animal models, still limits our understanding of the many actions and relative contributions of specific MMPs. Our future potential ability to control vascular remodeling via regulation of MMPs will also depend on reaching a consensus of what is indeed "good" or "bad" vascular remodeling, concepts that have continued to evolve and change. PMID- 11861413 TI - Mutations of the light meromyosin domain of the beta-myosin heavy chain rod in hypertrophic cardiomyopathy. AB - Familial hypertrophic cardiomyopathy (HCM) is caused by mutations in 9 sarcomeric protein genes. The most commonly affected is beta-myosin heavy chain (MYH7), where missense mutations cluster in the head and neck regions and directly affect motor function. Comparable mutations have not been described in the light meromyosin (LMM) region of the myosin rod, nor would these be expected to directly affect motor function. We studied 82 probands with HCM in whom no mutations had been found in MYH7 exons encoding the head and neck regions of myosin nor in the other frequently implicated disease genes. Primers were designed to amplify exons 24 to 40 of MYH7. These amplimers were subjected to temperature modulated heteroduplex analysis by denaturing high-performance liquid chromatography. An Ala1379Thr missense mutation in exon 30 segregated with disease in three families and was not present in 200 normal chromosomes. The mutation occurred on two haplotypes, indicating that it was not a polymorphism linked with another disease-causing mutation. The position of this residue within the LMM region of myosin suggests that it may be important for thick filament assembly or for accessory protein binding. A further missense mutation in exon 37, Ser1776Gly, segregated with disease in a single family and was absent from 400 population-matched control chromosomes. Because the Ser1776 residue occupies a core position in the myosin rod at which the substitution of glycine is extremely energetically unfavorable, it is likely to disrupt the coiled-coil structure. We conclude that mutation of the LMM can cause HCM and that such mutations may act through novel mechanisms of disease pathogenesis involving myosin filament assembly or interaction with thick filament binding proteins. PMID- 11861414 TI - Loss of SR-BI expression leads to the early onset of occlusive atherosclerotic coronary artery disease, spontaneous myocardial infarctions, severe cardiac dysfunction, and premature death in apolipoprotein E-deficient mice. AB - Murine models of atherosclerosis, such as the apolipoprotein E (apoE) or the LDL receptor knockout mice, usually do not exhibit many of the cardinal features of human coronary heart disease (CHD), eg, spontaneous myocardial infarction, severe cardiac dysfunction, and premature death. Here we show that mice with homozygous null mutations in the genes for both the high density lipoprotein receptor SR-BI and apoE (SR-BI/apoE double knockout [dKO] mice) exhibit morphological and functional defects with similarities to those seen in human CHD. When fed a standard chow diet, these hypercholesterolemic animals developed significant atherosclerotic lesions in the aortic sinus as early as 4 to 5 weeks after birth. We now show that they also exhibited extensive lipid-rich coronary artery occlusions and spontaneously developed multiple myocardial infarctions and cardiac dysfunction (eg, enlarged hearts, reduced ejection fraction and contractility, and ECG abnormalities). Their coronary arterial lesions, which were strikingly similar to human atherosclerotic plaques, exhibited evidence of cholesterol clefts and extensive fibrin deposition, indicating hemorrhage and clotting. All of the dKO mice died by 8 weeks of age (50% mortality at 6 weeks). Thus, SR-BI/apoE dKO mice provide a new murine model for CHD and may help better define the role of lipoprotein metabolism and atherosclerosis in the pathogenesis of myocardial infarction and cardiac dysfunction. Furthermore, these animals may be useful for preclinical testing of potential genetic and/or pharmacological therapies for CHD. PMID- 11861415 TI - Regression of atherosclerosis in monkeys reduces vascular superoxide levels. AB - Superoxide (O2*-) in arteries may contribute to atherosclerosis in part by inactivation of nitric oxide. We hypothesized that regression of atherosclerosis in nonhuman primates is associated with a decrease in vascular NAD(P)H oxidase, decreased O2*- levels, and improved endothelium-dependent relaxation. Cynomolgus monkeys (n=28) were fed an atherogenic diet for 47+/-10 (mean+/-SE) months. In carotid arteries (containing advanced lesions), femoral arteries (moderate lesions), and saphena arteries (minimal lesions), we examined O2*- levels and vasomotor function. Compared with vessels from normal monkeys (n=8), O2*- levels (measured by lucigenin-enhanced chemiluminescence) were 3.3-fold higher in carotid, 1.7-fold higher in femoral, and not different in saphena arteries from atherosclerotic monkeys. Dihydroethidium staining also demonstrated increased O2* levels throughout the vessel wall in femoral and carotid arteries from atherosclerotic monkeys. Components of the NAD(P)H oxidase (p22(phox) and p47(phox)) were increased in atherosclerotic arteries, and immunohistochemistry demonstrated colocalization primarily to areas of macrophage infiltration. Relaxation to acetylcholine was impaired in carotid and femoral, but not saphena, arteries from atherosclerotic monkeys. After 8 months of regression diet (n=9), serum cholesterol decreased to normal, and O2*- levels (basal and NAD(P)H stimulated), as well as expression of NAD(P)H oxidase, returned toward normal. Relaxation to acetylcholine improved in femoral arteries, but not in the more diseased carotid arteries. We conclude that, in a primate model of moderately severe atherosclerosis and regression of atherosclerosis, changes in endothelial function are inversely related to O2*- and NAD(P)H oxidase levels. Reduction in vascular O2*- during regression of atherosclerosis may contribute to improvement in vasomotor function. PMID- 11861416 TI - Bone marrow-derived endothelial progenitor cells participate in cerebral neovascularization after focal cerebral ischemia in the adult mouse. AB - We investigated whether circulating endothelial progenitor cells contribute to neovascularization after stroke. Donor bone marrow cells obtained from transgenic mice constitutively expressing beta-galactosidase transcriptionally regulated by an endothelial-specific promoter, Tie2, were injected into adult mice. Focal cerebral ischemia was induced by embolic middle cerebral artery (MCA) occlusion and changes of cerebral blood flow (CBF) were measured by perfusion-weighted magnetic resonance imaging (MRI). Laser scanning confocal microscopy (LSCM), immunohistochemistry and X-gal staining were performed. Perfusion-weighted MRI demonstrated increases in CBF around the boundary of an infarct area 1 month after ischemia. Morphological and 3-dimensional image analyses revealed enlarged and thin-walled blood vessels with sprouting or intussusception at the boundary of the ischemic lesion, which closely corresponded to elevated CBF areas detected on perfusion-weighted MRI, indicating the presence of neovascularization. X-gal and double immunostaining demonstrated that Tie2-lacZ-positive cells incorporated into sites of neovascularization at the border of the infarct, and these cells exhibited an endothelial antigenic marker (von Willebrand factor). In addition, bone marrow recipient mice without ischemia showed incorporation of Tie2-lacZ expressing cells into vessels of the choroid plexus. These data suggest that formation of new blood vessels in the adult brain after stroke is not restricted to angiogenesis but also involves vasculogenesis and that circulating endothelial progenitor cells from bone marrow contribute to the vascular substructure of the choroid plexus. PMID- 11861417 TI - Spatiotemporal transition to conduction block in canine ventricle. AB - Interruption of periodic wave propagation by the nucleation and subsequent disintegration of spiral waves is thought to mediate the transition from normal sinus rhythm to ventricular fibrillation. This sequence of events may be precipitated by a period doubling bifurcation, manifest as a beat-to-beat alternation, or alternans, of cardiac action potential duration and conduction velocity. How alternans causes the local conduction block required for initiation of spiral wave reentry remains unclear, however. In the present study, a mechanism for conduction block was derived from experimental studies in linear strands of cardiac tissue and from computer simulations in ionic and coupled maps models of homogeneous one-dimensional fibers. In both the experiments and the computer models, rapid periodic pacing induced marked spatiotemporal heterogeneity of cellular electrical properties, culminating in paroxysmal conduction block. These behaviors resulted from a nonuniform distribution of action potential duration alternans, secondary to alternans of conduction velocity. This link between period doubling bifurcations of cellular electrical properties and conduction block may provide a generic mechanism for the onset of tachycardia and fibrillation. PMID- 11861418 TI - Imbalance between xanthine oxidase and nitric oxide synthase signaling pathways underlies mechanoenergetic uncoupling in the failing heart. AB - Inhibition of xanthine oxidase (XO) in failing hearts improves cardiac efficiency by an unknown mechanism. We hypothesized that this energetic effect is due to reduced oxidative stress and critically depends on nitric oxide synthase (NOS) activity, reflecting a balance between generation of nitric oxide (NO) and reactive oxygen species. In dogs with pacing-induced heart failure (HF), ascorbate (1000 mg) mimicked the beneficial energetic effects of allopurinol, increasing both contractility and efficiency, suggesting an antioxidant mechanism. Allopurinol had no additive effect beyond that of ascorbate. Crosstalk between XO and NOS signaling was assessed. NOS inhibition with N(G)-monomethyl-L arginine (L-NMMA; 20 mg/kg) had no effect on basal contractility or efficiency in HF, but prevented the +26.2+/-3.5% and +66.5+/-17% enhancements of contractility and efficiency, respectively, observed with allopurinol alone. Similarly, improvements in contractility and energetics due to ascorbate were also inhibited by L-NMMA. Because of the observed NOS-XO crosstalk, we predicted that in normal hearts NOS inhibition would uncover a depression of energetics caused by XO activity. In normal conscious dogs, L-NMMA increased myocardial oxygen consumption (MVO2) while lowering left ventricular external work, reducing efficiency by 31.1+/-3.8% (P<0.005). Lowered efficiency was reversed by XO inhibition (allopurinol, 200 mg) or by ascorbate without affecting cardiac load or systemic hemodynamics. Single-cell immunofluorescence detected XO protein in cardiac myocytes that was enhanced in HF, consistent with autocrine signaling. These data show that both NOS and XO signaling systems participate in the regulation of myocardial mechanical efficiency and that upregulation of XO relative to NOS contributes to mechanoenergetic uncoupling in heart failure. PMID- 11861419 TI - The Na+-Ca2+ exchanger is essential for the action of cardiac glycosides. AB - The widely accepted model to explain the positive inotropic effect of cardiac glycosides invokes altered Na+-Ca2+ exchange activity secondary to Na+ pump inhibition. However, proof of this model is lacking and alternative mechanisms have been proposed. We directly tested the role of the Na+-Ca2+ exchanger in the action of the glycoside ouabain using Na+-Ca2+ exchanger knockout mice. Ablation of the exchanger is embryonic lethal, but contractility can be studied in embryonic heart tubes at day 9.5 postcoitum. Heart tubes isolated from homozygous Na+-Ca2+ exchanger knockout mice (NCX-/-) display surprisingly normal Ca2+ transients. Removal of extracellular Na+ induces Ca2+ overload in wild-type heart tubes but does not alter the Ca2+ transients of NCX-/- heart tubes. Similarly, ouabain, at levels causing Ca2+ overload in wild-type heart tubes, has no effect on NCX-/- heart tubes. We conclude that in embryonic mouse myocytes the Na+-Ca2+ exchanger is absolutely required for the effect of cardiac glycosides on Ca2+(i). PMID- 11861420 TI - Protein kinase A phosphorylation of the ryanodine receptor does not affect calcium sparks in mouse ventricular myocytes. AB - Ryanodine receptor (RyR) phosphorylation by protein kinase A (PKA) may be important in modulating resting sarcoplasmic reticulum (SR) Ca2+ release, especially in heart failure. However, clear cellular data on PKA-dependent modulation of cardiac RyRs is limited because of difficulty in distinguishing between PKA effects on RyR, phospholamban (PLB), and Ca2+ current. To clarify this, we measured resting Ca2+ sparks in streptolysin-O permeabilized ventricular myocytes from wild-type (WT) and PLB knockout (PLB-KO) mice and transgenic mice expressing only double-mutant PLB (PLB-DM) that lacks the regulatory phosphorylation sites (S16A/T17A). In WT myocytes, cAMP dramatically increased Ca2+ spark frequency (CaSpF) by 2- and 3-fold when [Ca2+] was clamped at 50 and 10 nmol/L (and the SR Ca2+ content also rose by 40% and 50%). However, in PLB-KO and PLB-DM, neither CaSpF nor SR Ca2+ load was changed by the addition of 10 micromol/L cAMP (even with phosphatase inhibition). PKA activation also increased Ca2+ spark amplitude, duration, and width in WT, but not in PLB-KO or PLB-DM. RyR phosphorylation was confirmed by measurements of 32P incorporation on immunoprecipitated RyR. In intact resting myocytes, PKA activation increased CaSpF 2.8-fold in WT, but not in PLB-KO, confirming results in permeabilized myocytes. We conclude that the PKA-dependent increase in myocyte CaSpF and size is entirely attributable to PLB phosphorylation and consequent enhanced SR Ca2+ load. PKA does not seem to have any appreciable effect on resting RyR function in these ventricular myocytes. Moreover, the data provide compelling evidence that elevated intra-SR [Ca2+] increases RyR gating independent of cytosolic [Ca2+] (which was clamped). PMID- 11861421 TI - Increased association of ZO-1 with connexin43 during remodeling of cardiac gap junctions. AB - The intercellular geometry of connexin43 (Cx43) gap junctional coupling is key to coordinated spread of electrical activation through the ventricle of the mammalian heart. A progressive redistribution of electrical and mechanical junctions into intercalated discs occurs during postnatal development. Breakdown of disc-localized pattern in the adult heart, to recapitulate immature distributions, is thought to be key to the genesis of conduction disturbance and arrhythmia. Recently, ZO-1 (a PDZ-MAGUK protein), has been suggested to have a role in generating coupling geometries between myocytes. We therefore investigated the codistribution of ZO-1 with Cx43 and N-cadherin in the adult rat ventricle using quantitative immunoconfocal and immunoelectron microscopy. These analyses indicated that, whereas ZO-1 and Cx43 codistribute within discs, only low to moderate point-by-point colocalization of Cx43 and ZO-1 is found within these domains compared with the relatively high level of colocalization between N cadherin and ZO-1. By contrast, levels of association between Cx43 and ZO-1 increased rapidly and significantly (P<0.001) after partial or complete enzymatic dissociation of myocytes from intact ventricle--a treatment known to induce gap junction endocytosis. Coimmunoprecipitation using Cx43- and ZO-1-specific antibodies confirmed that significantly (P<0.03) increased ZO-1 is precipitated relative to Cx43 in freshly dissociated myocytes as compared with intact ventricle. On immunoblots, decreases in Cx43 relative mobility, consistent with increased phosphorylation, were observed following myocyte dissociation. The increased ZO-1-Cx43 association that occurs after remodeling of myocyte intercellular contacts indicates the possibility of unanticipated roles for ZO-1 in gap junction turnover during cardiac development and disease processes. PMID- 11861422 TI - Sphingosine-1-phosphate, a platelet-derived lysophospholipid mediator, negatively regulates cellular Rac activity and cell migration in vascular smooth muscle cells. AB - Previous studies demonstrated that sphingosine-1-phosphate (S1P) induced migration of human umbilical vein endothelial cells (HUVECs) whereas it inhibited that of vascular smooth muscle cells (SMCs). This study explored the molecular mechanisms underlying the contrasting S1P actions on vascular cell motility. In rat and human aortic SMCs, the chemoattractant platelet-derived growth factor B chain (PDGF) induced rapid 5- to 6-fold increases in the cellular amount of GTP bound, active form of Rac. S1P did not affect PDGF-stimulated tyrosine phosphorylation of PDGF-beta receptor, but strongly inhibited PDGF-induced Rac activation, with a dose-response relationship similar to that for inhibition of PDGF-elicited chemotaxis. Dihydrosphingosine-1-phosphate, which is a weaker agonist for the S1P receptors, but not an inactive ligand sphingosine, also inhibited PDGF-stimulated chemotaxis and Rac activation although to lesser extents compared with S1P, suggesting that negative regulation by S1P of both chemotaxis and Rac was a receptor-mediated process. In contrast, S1P by itself stimulated Rac activity in HUVECs. Among the five S1P receptor isoforms, SMCs prominently expressed Edg-5 mRNA, whereas HUVECs expressed abundant Edg-1 mRNA but lacked detectable expression of Edg-5 mRNA. Adenovirus-mediated expression of a dominant-negative form of either Rac or Cdc42, but not RhoA, markedly attenuated chemotaxis of SMCs and HUVECs toward PDGF and S1P, respectively. Overexpression of Edg-1 in SMCs and Edg-5 in HUVECs reduced S1P-induced inhibition and stimulation, respectively, of Rac activity and migration. These results together indicate that Edg isoform-specific, negative or positive regulation of cellular Rac activity is critically involved in S1P-mediated bimodal regulation of cell motility in SMCs and HUVECs. PMID- 11861423 TI - Vitamin E oxidation in human atherosclerotic lesions. AB - Oxidation of low-density lipoproteins (LDL) is a key process in atherogenesis, and vitamin E (alpha-tocopherol, TOH) has received attention for its potential to attenuate the disease. Despite this, the type and extent of TOH oxidation and its relationship to lipid oxidation in the vessel wall where lesions develop remain unknown. Therefore, we measured oxidized lipids, TOH, and its oxidation products, alpha-tocopherylquinone (TQ), 2,3- and 5,6-epoxy-alpha-tocopherylquinones by gas chromatography-mass spectrometry analysis in human lesions representing different stages of atherosclerosis. We also oxidized LDL in vitro to establish "footprints" of TOH oxidation product for different oxidants. The in vitro studies demonstrated that tocopherylquinone epoxides are the major products when LDL is exposed to the one-electron (ie, radical) oxidants, peroxyl radicals, and copper ions, whereas TQ preferentially accumulates with the two-electron (nonradical) oxidants, hypochlorite, and peroxynitrite. In human lesions, the relative extent of TOH oxidation was maximal early in the disease where it exceeded lipid oxidation. Independent of the disease stage, TQ was always the major oxidation product with all products together representing <20% of the total TOH present, and the oxidation product profile mirroring that formed during LDL oxidation by activated monocytes in the presence of nitrite. In contrast, oxidized lipid increased with increasing disease severity. These results suggest that two-electron oxidants are primarily responsible for TOH oxidation in the artery wall, and that the extent of TOH oxidation is limited yet substantial lipid oxidation takes place. This study may have important implications regarding antioxidant supplements aimed at preventing LDL oxidation and hence atherogenesis. PMID- 11861424 TI - A role for the beta-catenin/T-cell factor signaling cascade in vascular remodeling. AB - Beta-catenin and T cell factor (Tcf) are distal components of the highly conserved Wnt pathway that govern cell fate and proliferation in lower organisms. Thus, we hypothesized that the regulation of beta-catenin and Tcf played a critical role in vascular remodeling. The first objective was to define beta catenin expression in vascular smooth muscle cells (VSMCs) after balloon injury. Indeed, beta-catenin mRNA and protein were significantly elevated 7 days after balloon injury in the rat carotid artery. We hypothesized that beta-catenin accumulation in response to vascular injury inhibited VSMC apoptosis. In line with our hypothesis, transfection of a degradation-resistant beta-catenin transgene into rat VSMCs significantly inhibited apoptosis. Accumulation of beta catenin also resulted in a 10-fold increase in the activation of Tcf. To test if Tcf was necessary to confer beta-catenin-induced survival, loss of function studies were carried out with a dominant negative Tcf-4 transgene lacking the beta-catenin binding domain, Tcf4(N31). Indeed, loss of Tcf-4 activity abolished beta-catenin-induced survival. We further postulated that beta-catenin and Tcf promoted cell cycle progression by activating cyclin D1, a target gene of Tcf-4. Beta-catenin activated cyclin D1, and this activation was partially blocked with loss of Tcf-4. In parallel, blockade of Tcf-4 resulted in inhibition of [3H]thymidine incorporation and partial blockade of the G1-S phase transition. In conclusion, beta-catenin and Tcf-4 play a dual role in vascular remodeling by inhibiting VSMC apoptosis and promoting proliferation. PMID- 11861425 TI - PPARgamma ligand inhibits osteopontin gene expression through interference with binding of nuclear factors to A/T-rich sequence in THP-1 cells. AB - Peroxisome proliferator-activated receptor gamma (PPARgamma) is a member of the nuclear receptor superfamily that acts as a key player in adipocyte differentiation, glucose metabolism, and macrophage differentiation. Osteopontin (OPN), a component of extracellular matrix, is elevated during neointimal formation in the vessel wall and is synthesized by macrophages in atherosclerotic plaques. In the present study, we investigated the molecular mechanisms regulating OPN gene expression by PPARgamma in THP-1 cells, a cell line derived from human monocytic leukemia cells. Northern and Western blot analyses showed that exposure of THP-1 cells to PMA (phorbol 12-myristate 13-acetate) increases OPN mRNA and protein levels in a time-dependent manner. PMA-induced OPN expression was significantly decreased by troglitazone (Tro) and other PPARgamma ligands. Transient transfection assays of the human OPN promoter/luciferase construct showed that PPARgamma represses OPN promoter activity, and the PPARgamma-responsive region within the OPN promoter lies between -1000 and -970 relative to the transcription start site. Site-specific mutation analysis and electrophoretic mobility shift assays indicated that a homeobox-like A/T-rich sequence between -990 and -981, which functions as a binding site for PMA-induced nuclear factors other than PPARgamma, mediates the repression of OPN expression by Tro. Furthermore, concatenated A/T-rich sequences conferred the PPARgamma responsiveness on the heterologous promoter. Taken together, these data suggest that PPARgamma ligand inhibits OPN gene expression through the interference with the binding of nuclear factors to A/T-rich sequence in THP-1 cells. PMID- 11861426 TI - PPARgamma but not PPARalpha ligands are potent repressors of major histocompatibility complex class II induction in atheroma-associated cells. AB - Peroxisome proliferator-activated receptors (PPARs) are essential in glucose and lipid metabolism and are implicated in metabolic disorders predisposing to atherosclerosis, such as diabetes and dyslipidemia. Conversely, antidiabetic glitazones and hypolipidemic fibrate drugs, known as PPARgamma and PPARalpha ligands, respectively, reduce the process of atherosclerotic lesion formation, which involves chronic immunoinflammatory processes. Major histocompatibility complex class II (MHC-II) molecules, expressed on the surface of specialized cells, are directly involved in the activation of T lymphocytes and in the control of the immune response. Interestingly, expression of MHC-II has recently been observed in atherosclerotic plaques, and it can be induced by the proinflammatory cytokine interferon-gamma (IFN-gamma) in vascular cells. To explore a possible role for PPAR ligands in the regulation of the immune response, we investigated whether PPAR activation affects MHC-II expression in atheroma-associated cells. In the present study, we demonstrate that PPARgamma but not PPARalpha ligands act as inhibitors of IFN-gamma-induced MHC-II expression and thus as repressors of MHC-II-mediated T-cell activation. All different types of PPARgamma ligands tested inhibit MHC-II. This effect of PPARgamma ligands is due to a specific inhibition of promoter IV of CIITA and does not concern constitutive expression of MHC-II. Thus, the beneficial effects of antidiabetic PPARgamma activators on atherosclerotic plaque development may be partly explained by their repression of MHC-II expression and subsequent inhibition of T-lymphocyte activation. PMID- 11861427 TI - S-nitrosoalbumin plasma levels in health and disease: facts or artifacts? Value of analytical chemistry in nitric oxide clinical research. PMID- 11861428 TI - Fabrication of pulsatile cardiac tissue grafts using a novel 3-dimensional cell sheet manipulation technique and temperature-responsive cell culture surfaces. AB - Recent progress in cell transplantation therapy to repair impaired hearts has encouraged further attempts to bioengineer 3-dimensional (3-D) heart tissue from cultured cardiomyocytes. Cardiac tissue engineering is currently pursued utilizing conventional technology to fabricate 3-D biodegradable scaffolds as a temporary extracellular matrix. By contrast, new methods are now described to fabricate pulsatile cardiac grafts using new technology that layers cell sheets 3 dimensionally. We apply novel cell culture surfaces grafted with temperature responsive polymer, poly(N-isopropylacrylamide) (PIPAAm), from which confluent cells detach as a cell sheet simply by reducing temperature without any enzymatic treatments. Neonatal rat cardiomyocyte sheets detached from PIPAAm-grafted surfaces were overlaid to construct cardiac grafts. Layered cell sheets began to pulse simultaneously and morphological communication via connexin43 was established between the sheets. When 4 sheets were layered, engineered constructs were macroscopically observed to pulse spontaneously. In vivo, layered cardiomyocyte sheets were transplanted into subcutaneous tissues of nude rats. Three weeks after transplantation, surface electrograms originating from transplanted grafts were detected and spontaneous beating was macroscopically observed. Histological studies showed characteristic structures of heart tissue and multiple neovascularization within contractile tissues. Constructs transplanted into 3-week-old rats exhibited more cardiomyocyte hypertrophy and less connective tissue than those placed into 8-week-old rats. Long-term survival of pulsatile cardiac grafts was confirmed up to 12 weeks. These results demonstrate that electrically communicative pulsatile 3-D cardiac constructs were achieved both in vitro and in vivo by layering cardiomyocyte sheets. Cardiac tissue engineering based on this technology may prove useful for heart model fabrication and cardiovascular tissue repair. The full text of this article is available at http://www.circresaha.org. PMID- 11861429 TI - Can measurement of apolipoprotein B replace the lipid profile in the follow-up of patients with lipoprotein disorders? PMID- 11861430 TI - Point-of-care screening for chromosomal anomalies in the first trimester of pregnancy. PMID- 11861431 TI - Unexpected suppression of immunoassay results by cross-reactivity: now a demonstrated cause for concern. PMID- 11861432 TI - Measuring and interpreting holo-transcobalamin (holo-transcobalamin II). PMID- 11861433 TI - Synthetic peptides identified from phage-displayed combinatorial libraries as immunodiagnostic assay surrogate quality-control targets. AB - BACKGROUND: Quantitative immunohistochemical (IHC) assays currently lack optimal reference quality-control material for cellular protein targets. To address this problem, we identified peptides that mimic the site on the native analyte to which the primary (monoclonal) antibody binds and used them as surrogate peptide controls. METHODS: We identified peptide candidates from a combinatorial peptide phage-display library that mimic the epitope for the 1D5 estrogen receptor (ER) monoclonal antibody (mAb). The peptide inserts of the phage clones were sequenced. Several phage-encoded peptides were then synthesized and analyzed for affinity and specificity. RESULTS: We identified phage clones that specifically bound to the ER 1D5 mAb. The binding was specific, in that the phage clones did not bind to two other isotype-matched mAbs. Their ability to bind the ER 1D5 mAb was related to the presence of a consensus sequence. Binding analysis revealed a K(d) of 8.3 x 10(-8) mol/L. The peptide was not recognized by any of 15 other mAbs commonly used for clinical IHC testing. Moreover, the peptide was able to inhibit the binding of ER 1D5 mAb to native ER, indicating that the peptide bound to ER 1D5 mAb at or close to the antigen-binding site. CONCLUSIONS: Surrogate peptide controls behave like the native analyte in terms of affinity and specificity. This technology may be especially useful when the native analyte is in short supply. PMID- 11861434 TI - Predominant hematopoietic origin of cell-free DNA in plasma and serum after sex mismatched bone marrow transplantation. AB - BACKGROUND: Despite current interest in the biology and diagnostic applications of cell-free DNA in plasma and serum, the cellular origin of this DNA is poorly understood. We used a sex-mismatched bone marrow transplantation model to study the relative contribution of hematopoietic and nonhematopoietic cells to circulating DNA. METHODS: We studied 22 sex-mismatched bone marrow transplantation patients. Paired buffy coat and plasma samples were obtained from all 22 patients. Matching serum samples were also obtained from seven of them. Plasma DNA, serum DNA, and buffy coat were quantified by real-time PCR of the SRY and beta-globin gene DNA. To investigate the effects of blood drawing and other preanalytical variables on plasma DNA concentrations, blood samples were also collected from 14 individuals who had not received transplants. The effects of blood sampling by syringe and needle, centrifugation, and time delay in blood processing were studied. RESULTS: The median percentage of Y-chromosome DNA in the plasma in female patients receiving bone marrow from male donors (59.5%) differed significantly (P <0.001) from that in the male patients receiving bone marrow from female donors (6.9%). This indicated that plasma DNA in the bone marrow transplantation recipients was predominantly of donor origin. Compared with paired plasma samples, serum samples had a median 14-fold higher DNA concentration, with the additional DNA being of donor origin. Control experiments indicated that none of the three tested preanalytical variables contributed to a significant change in cell-free DNA concentration. CONCLUSIONS: After bone marrow transplantation, the DNA in plasma and serum is predominantly hematopoietic in origin. Apart from the biological implications of this observation, this finding suggests that plasma and serum can be used as alternative materials for the study of postbone marrow transplantation chimerism. PMID- 11861435 TI - Diagnostic biochip array for fast and sensitive detection of K-ras mutations in stool. AB - BACKGROUND: Tumor cells that shed into stool are attractive targets for molecular screening and early detection of colon or pancreatic malignancies. We developed a diagnostic test to screen for 10 of the most common mutations of codons 12 and 13 of the K-ras gene by hybridization to a new biochip array. METHODS: DNA was isolated from 26 stool samples by column-based extraction from 9 cell lines. Peptide nucleic acid (PNA)-mediated PCR clamping was used for mutant-specific amplification. We used a biochip, consisting of a small plastic support with covalently immobilized 13mer oligonucleotides. The read out of the biochip was done by confocal time-resolved laser scanning. Hybridization, scanning, and data evaluation could be performed in <2 h. RESULTS: Approximately 80 ng of DNA was obtained from 200-mg stool samples. No inhibition of the PCR by remaining impurities from stool was observed. Mutation detection was possible in 1000-fold excess of wild-type sequence. Discrimination ratios between the mutations were >19 as demonstrated by hybridization with tumor cell line DNA. Stool samples (n = 26) were analyzed in parallel with PNA-PCR, restriction assay for K-ras codon 12 mutations, sequencing, and hybridization to the biochip. Nine mutations were found by hybridization, all confirmed by sequencing. PNA-PCR alone leads to an overestimation of mutations because suppression of the wild type is not effective enough with high concentrations of wild-type DNA. The restriction assay found only four mutations. CONCLUSIONS: The K-ras biochip is well suited for fast mutation detection from stool in colorectal cancer screening. PMID- 11861436 TI - Guidelines and recommendations for laboratory analysis in the diagnosis and management of diabetes mellitus. AB - BACKGROUND: Multiple laboratory tests are used in the diagnosis and management of patients with diabetes mellitus. The quality of the scientific evidence supporting the use of these assays varies substantially. APPROACH: An expert committee drafted evidence-based recommendations for the use of laboratory analysis in patients with diabetes. An external panel of experts reviewed a draft of the guidelines, which were modified in response to the reviewers' suggestions. A revised draft was posted on the Internet and was presented at the AACC Annual Meeting in July, 2000. The recommendations were modified again in response to oral and written comments. The guidelines were reviewed by the Professional Practice Committee of the American Diabetes Association. CONTENT: Measurement of plasma glucose remains the sole diagnostic criterion for diabetes. Monitoring of glycemic control is performed by the patients, who measure their own plasma or blood glucose with meters, and by laboratory analysis of glycated hemoglobin. The potential roles of noninvasive glucose monitoring, genetic testing, autoantibodies, microalbumin, proinsulin, C-peptide, and other analytes are addressed. SUMMARY: The guidelines provide specific recommendations based on published data or derived from expert consensus. Several analytes are of minimal clinical value at the present time, and measurement of them is not recommended. PMID- 11861437 TI - Point-of-care time-resolved immunofluorometric assay for human pregnancy associated plasma protein A: use in first-trimester screening for Down syndrome. AB - BACKGROUND: Screening for Down syndrome in the first trimester by a combination of fetal nuchal translucency thickness and maternal serum pregnancy-associated plasma protein A (PAPP-A) and free beta-human chorionic gonadotropin has been shown to be effective and efficient. We aimed to develop a fast point-of-care assay that could be placed in one-stop clinics for the measurement of PAPP-A. METHODS: We developed a two-site, one-step assay that uses two monoclonal antibodies (mAbs) to PAPP-A, based on a dry-reagent, all-in-one immunoassay concept with a stable fluorescent lanthanide chelate and time-resolved fluorometry. One antibody (mAb 10E1) was biotinylated, and the other (mAb 234-5) was europium-labeled, both via the epsilon-amino groups of surface lysine residues. The assay was performed on an AIO immunoanalyzer at 36 degrees C in single, streptavidin-coated microtitration wells that contained the dry reagents. PAPP-A, either in free or complexed form, was detected by the antibodies used. RESULTS: The assay procedure required 20 min and used 10 microL of sample. The calibration curve was linear from 5 to 10 000 mIU/L. The detection limit was 0.5 mIU/L. Intra- and interassay imprecision (CV) was < or = 4.3% and 8.3%, respectively, for whole blood, plasma, or serum samples. Recovery was 93-96% for serum, 95-108% for heparin-derived whole blood, and 98-103% for heparin-derived plasma. Parallelism was observed in all three matrices. Results correlated [slope = 0.85 (confidence interval, 0.82-0.87); intercept = -33 (confidence interval, 58 to -9); S(y:x) = 85 mIU/L; r = 0.991; n = 100] with those obtained by a Delfia assay. Heparin did not affect the assay, but EDTA markedly reduced PAPP-A values. PAPP-A was stable at 4 degrees C for at least 18 days in serum and for 8 days in heparin-derived whole blood or plasma. CONCLUSIONS: The present assay appears suited for use in one-stop clinics for screening for Down syndrome in the first trimester, with results available within 1 h. PMID- 11861438 TI - Can measurement of serum apolipoprotein B replace the lipid profile monitoring of patients with lipoprotein disorders? AB - BACKGROUND: Current clinical guidelines require that five indices (total cholesterol, LDL cholesterol, HDL cholesterol, triglycerides, and the total/HDL cholesterol ratio) be measured or calculated to assess the lipid-related risk of vascular disease. All five are also targets of therapy and therefore all must be measured initially and at follow-up. Considerable evidence indicates that apolipoprotein B (apo B) is a better index of reaching or not reaching treatment targets than total or LDL cholesterol. METHODS: The objective of this study was to examine whether measurement of a single marker (apo B) led to the same categorization of risk as the traditional five indices (lipid profile). If both apo B and lipid profile indicated that the patient was either within or outside their respective treatment targets, the indices were considered concordant. If not, the indices were considered discordant. Concordance/discordance was examined in 215 patients at their first and last clinic visit. RESULTS: Concordance was high in both higher (88% at the first and 92% at the last clinic visit) and lower (76% at the first and 78% at the last clinic visit) risk groups at both the initial and final visits. Discordance was virtually restricted to the group with hypertriglyceridemia with normal concentrations of apo B, a group in which little independent evidence points to any substantially increased risk of vascular disease. CONCLUSIONS: These data raise the possibility that at least for high risk patients treated with statins, follow-up could be simplified and expenses reduced if only apo B were measured. They also raise the possibility that outcome might be improved if the therapeutic algorithm were simplified. PMID- 11861439 TI - Performance of four homogeneous direct methods for LDL-cholesterol. AB - BACKGROUND: Homogeneous LDL-cholesterol methods from Genzyme, Reference Diagnostics, Roche, and Sigma were evaluated for precision, accuracy, and specificity for LDL in the presence of abnormal lipoproteins. METHODS: Each homogeneous method was performed by a Roche/Hitachi 911 according to the vendors' instructions, and the results were compared with the beta-quantification reference method. We measured precision over 20 days using quality-control and frozen serum specimens. Sera from 100 study participants, including 60 with hyperlipidemias, were assayed by each method. Accuracy was evaluated from regression and total error analysis. Specificity was evaluated from the bias (as a percentage) vs concentration of triglycerides. RESULTS: The total CV was <2% for all methods. Regression slope and intercept (with 95% confidence intervals) were as follows: Genzyme, 0.955 (0.92 to 0.99) and 30.3 (-12 to 73) mg/L; Reference Diagnostics, 0.975 (0.93 to 1.02) and -8 (-63 to 47) mg/L; Roche, 1.067 (1.02 to 1.11) and -101 (-161 to -42) mg/L; and Sigma, 0.964 (0.91 to 1.02) and 164 (89 to 239) mg/L. The percentages of individual results with >12% bias were as follows: Genzyme, 8.0%; Reference Diagnostics, 11.0%; Roche, 10.0%; and Sigma, 30.0%. Total error calculated from mean systematic bias and all-sources random bias was as follows: Genzyme, 12.6%; Reference Diagnostics, 16.5%; Roche, 41.6%; and Sigma, 38.3%. Slopes of bias (as a percentage) vs triglycerides were P <0.001 for all methods except the Roche method, which was P = 0.094. CONCLUSIONS: The evaluated methods show nonspecificity toward abnormal lipoproteins, thus compromising their ability to satisfy the National Cholesterol Education Program goal for a total error of <12%. These homogeneous LDL-cholesterol results do not improve on the performance of LDL-cholesterol calculated by the Friedewald equation at triglyceride concentrations <4000 mg/L. PMID- 11861440 TI - Reagent-free, simultaneous determination of serum cholesterol in HDL and LDL by infrared spectroscopy. AB - BACKGROUND: The purpose of this study was to assess the feasibility of infrared (IR) spectroscopy for the simultaneous quantification of serum LDL-cholesterol (LDL-C) and HDL-cholesterol (HDL-C) concentrations. METHODS: Serum samples (n = 90) were obtained. Duplicate aliquots (5 microL) of the serum specimens were dried onto IR-transparent barium fluoride substrates, and transmission IR spectra were measured for the dry films. In parallel, the HDL-C and LDL-C concentrations were determined separately for each specimen by standard methods (the Friedewald formula for LDL-C and an automated homogeneous HDL-C assay). The proposed IR method was then developed with a partial least-squares (PLS) regression analysis to quantitatively correlate IR spectral features with the clinical analytical results for 60 randomly chosen specimens. The resulting quantification methods were then validated with the remaining 30 specimens. The PLS model for LDL-C used two spectral ranges (1700-1800 and 2800-3000 cm(-1)) and eight PLS factors, whereas the PLS model for HDL-C used three spectral ranges (800-1500, 1700-1800, and 2800-3500 cm(-1)) with six factors. RESULTS: For the 60 specimens used to train the IR-based method, the SE between IR-predicted values and the clinical laboratory assays was 0.22 mmol/L for LDL-C and 0.15 mmol/L for HDL-C (r = 0.98 for LDL-C; r = 0.91 for HDL-C). The corresponding SEs for the test spectra were 0.34 mmol/L (r = 0.96) and 0.26 mmol/L (r = 0.82) for LDL-C and HDL-C, respectively. The precision for the IR-based assays was estimated by the SD of duplicate measurements to be 0.11 mmol/L (LDL-C) and 0.09 mmol/L (HDL-C). CONCLUSIONS: IR spectroscopy has the potential to become the clinical method of choice for quick and simultaneous determinations of LDL-C and HDL-C. PMID- 11861441 TI - Digoxin assays: frequent, substantial, and potentially dangerous interference by spironolactone, canrenone, and other steroids. AB - BACKGROUND: A case of digoxin toxicity resulted from falsely low values with the MEIA II assay for digoxin (AxSYM; Abbott). The low results were caused by negative interference from canrenone and spironolactone, the latter of which has recently been advocated for the treatment of severe heart failure. Analytical interference from spironolactone has been reported, but little information is available for this effect with newer digoxin assays. METHODS: We examined nine assays (AxSYM, IMx, TDx, Emit, Dimension, aca, TinaQuant, Elecsys, and Vitros for interference by spironolactone, canrenone, and three metabolites. Additionally, all routine digoxin measurements (AxSYM) over a period of 16.5 months (n = 3089) were monitored for interference. RESULTS: Suppression of the expected values by canrenone (3125 microg/L) was observed for the AxSYM (42% of expected value), IMx (51%), and Dimension (78%) assays. A positive bias was observed for the aca (0.7 microg/L), the TDx (0.62 microg/L), and the Elecsys (>0.58 microg/L). Twenty-five of 669 routinely monitored patients had falsely low results. Nineteen of these had potentially toxic concentrations of digoxin (Emit; >2.0 microg/L), although the AxSYM assay indicated therapeutic or less severe toxic concentrations (Delta(max) = 7.1 microg/L). Except for two unresolved cases, this was attributable to spironolactone, canrenone, hydrocortisone, or prednisolone. Standard doses of spironolactone (up to 50 mg/day) in patients with heart failure displayed inhibition <11%. CONCLUSIONS: The frequency and magnitude of the false negative results particularly compromise the use of both microparticle enzyme immunoassays. Not only may toxic concentrations remain unidentified, but intoxication could occur should dosage be increased because of falsely low results. With 11 million digoxin tests/year ordered in the US, conceivably many patients could be adversely affected. PMID- 11861442 TI - Comparison of the Emit immunoassay with HPLC for therapeutic drug monitoring of mycophenolic acid in pediatric renal-transplant recipients on mycophenolate mofetil therapy. AB - BACKGROUND: HPLC is currently the preferred method for accurate measurement of mycophenolic acid (MPA). This study was designed to validate the Emit compared with HPLC in relation to clinical outcome measurements. METHODS: Pediatric renal transplant recipients (n = 50) on an immunosuppressive triple regimen consisting of cyclosporin A, prednisone, and mycophenolate mofetil (600 mg/m(2) twice per day) were investigated in an open-label prospective study. Pharmacokinetic profiles over 12 h were obtained at 1 week, 3 weeks, 3 months, and 6 months posttransplant. Plasma MPA was measured by both reversed-phase HPLC and the Emit immunoassay. RESULTS: There was an association between the risk of acute rejection episodes and low area under the curve values from t(0) to t(12h) (AUC(0 12)) for MPA (MPA-AUC(0-12)) or predose concentrations of MPA derived from both HPLC and Emit measurements. According to ROC analysis, an AUC value of 33.8 mg x h/L for MPA from t(0) to t(12h) (MPA-AUC(0-12)) determined by HPLC had a diagnostic sensitivity of 80% and a diagnostic specificity of 57%. The corresponding value of the Emit was 36.1 mg x h/L. For the predose concentration (MPA-c(12)), a concentration of 1.2 mg/L determined by HPLC and 1.4 mg/L determined by Emit gave a sensitivity of 80% and a specificity of 60%, respectively. There was no association of any pharmacokinetic variables derived from total MPA measurements with an increased risk of side effects related to mycophenolate mofetil. CONCLUSIONS: The Emit assay appears to have a comparable diagnostic efficacy to HPLC for assessing the risk of acute rejection in pediatric renal-transplant recipients. However, because of the cross-reactivity of the antibody used in the Emit assay with the active MPA acyl glucuronide metabolite, the decision thresholds for the Emit were higher than those calculated from HPLC measurements. PMID- 11861443 TI - Direct assay for cobalamin bound to transcobalamin (holo-transcobalamin) in serum. AB - BACKGROUND: Only cobalamin carried by transcobalamin (holo-transcobalamin) is available for cellular uptake and hence is physiologically relevant. However, no reliable or accurate methods for quantifying holo-transcobalamin are available. We report a novel holo-transcobalamin assay based on solid-phase capture of transcobalamin. METHODS: A monoclonal antibody specific for human transcobalamin with an affinity constant >10(10) L/mol was immobilized on magnetic microspheres to capture and concentrate transcobalamin. The cobalamin bound to transcobalamin was then released and assayed by a competitive binding radioassay. The quantification of holo-transcobalamin was accomplished using calibrators composed of recombinant, human holo-transcobalamin. RESULTS: The assay was specific for holo-transcobalamin and had a detection limit of 5 pmol/L. Within-run and total imprecision (CV) was 5% and 8-9%, respectively. The working range (CV <20%) was 5 370 pmol/L. Dilutions of serum were linear in the assay range. The recovery of recombinant, human holo-transcobalamin added to serum was 93-108%. A 95% reference interval of 24-157 pmol/L was established for holo-transcobalamin in 105 healthy volunteers 20-80 years of age. For 72 of these sera, holo-haptocorrin and total cobalamin were also determined. Whereas holo-haptocorrin correlated well (r(2) = 0.87) with total cobalamin, holo-transcobalamin correlated poorly (r(2) = 0.23) with total cobalamin or holo-haptocorrin. CONCLUSIONS: The solid phase capture assay provides a simple, reliable method for quantitative determination of holo-transcobalamin in serum. PMID- 11861444 TI - Validation of liquid chromatography-tandem mass spectrometry method for analysis of urinary conjugated metanephrine and normetanephrine for screening of pheochromocytoma. AB - BACKGROUND: Metanephrines are biochemical markers for tumors of the adrenal medulla (e.g., pheochromocytoma) and other tumors derived from neural crest cells (e.g., paragangliomas and neuroblastomas). We describe a liquid chromatography tandem mass spectrometry (LC-MS/MS) method for the measurement of urinary conjugated metanephrines. METHODS: We added 250 ng of d3-metanephrine (d3-MN) and 500 ng of d3-normetanephrine (d3-NMN) to 1 mL of urine samples as stable isotope internal standards. The samples were then acidified, hydrolyzed for 20 min in a 100 degree C water bath, neutralized, and prepared by solid-phase extraction. The methanol eluates were analyzed by LC-MS/MS in the selected-reaction-monitoring mode after separation on a reversed-phase amide C16 column. RESULTS: Multiple calibration curves for the analysis of urine MN and NMN exhibited consistent linearity and reproducibility in the range of 10-5000 microg/L. Interassay CVs were 5.7-8.6% at mean concentrations of 90-4854 microg/L for MN and NMN. The detection limit was 10 microg/L. Recovery of MN and NMN (144-2300 microg/L) added to urine was 91-114%. The regression equation for the LC-MS/MS (x) and colorimetric (y) methods was: y = 0.81x - 0.006 (r = 0.822; n = 110). The equation for the HPLC (x) and LC-MS/MS (y) methods was: y = 1.09x + 0.05 (r = 0.998; n = 40). CONCLUSIONS: The sensitivity and specificity of the MS/MS method for urinary conjugated metanephrines offer advantages over colorimetric, immunoassay, HPLC, and gas chromatography-mass spectrometry methods because of elimination of drug interferences, high throughput, and short chromatographic run time. PMID- 11861445 TI - Evaluation of an automated preanalytical robotic workstation at two academic health centers. AB - BACKGROUND: Purchase of automated systems in today's clinical laboratory needs justification based on demonstrable improvements in efficiency and a sound payback model. Few studies provide information on laboratory automation that focuses on the preanalytical portion of specimen processing. METHODS: We recently evaluated an automated preanalytical processing unit (GENESIS FE500) at two academic health centers. This preanalytical unit processes blood specimens through automated specimen sorting, centrifugation, decapping, labeling, aliquoting, and placement of the processed specimen in the analytical rack. We quantified the output of the FE500 by processing >3000 barcode-labeled specimens according to a protocol designed to test all of the features of this automated specimen-processing unit. RESULTS: Depending on the batch size, aliquot number requested, and percentage of tubes that required centrifugation, the mean system output performance varied between 93 and 502 total tubes/h. Throughput increased when the batch size expanded from 40 or 100 samples (mean = 211 total tubes processed/h) to batch sizes of 200 and 300 tubes (mean = 474 total tube processed/h). The GENESIS FE500 processed specimen tubes differing in size from 13 x 65 mm (width x height) to 16 x 100. At one site, the FE500 was operated by one person, compared with the three individuals required to perform the same tasks manually. Finally, the specimen-processing error rate determined at one of the institutions was significantly reduced. CONCLUSIONS: We conclude that the GENESIS FE500 effectively reduces the labor associated with specimen processing; decreases the number of laboratory errors that occur with specimen sorting, labeling, and aliquoting; and improves the integrity of specimen handling throughout the steps of specimen processing. PMID- 11861446 TI - Determination of anti-acetylcholine receptor antibodies in myasthenic patients by use of time-resolved fluorescence. AB - BACKGROUND: Autoantibodies against nicotinic acetylcholine receptor (nAChR) in myasthenia gravis (MG) patients are usually detected by radioimmunoprecipitation assays using extracted acetylcholine receptors labeled irreversibly with 125I alpha-bungarotoxin (alpha-BuTx). To provide a nonradioactive immunoassay, we established an assay using nAChRs labeled with Eu(3+)-alpha-cobratoxin (alpha CTx). METHODS: We derivatized alpha-CTx with a diethylenetriaminepentaacetate moiety and formed a complex with Eu(3+). The complex was purified by HPLC, and the fractions were tested for binding to Torpedo and human nAChRs. The most active fractions were used to label nAChRs for the immunoprecipitation assay, and the bound Eu(3+) was quantified by time-resolved fluorescence. RESULTS: Eu(3+) labeled alpha-CTx competed with 125I-alpha-BuTx for binding to Torpedo nAChRs and saturated the binding sites of human nAChRs, with a K(d) of 7.2 x 10(-9) mol/L. Results of the immunoassay performed with Eu(3+)-labeled alpha-CTx were similar to those obtained with 125I-alpha-BuTx, with a slightly higher limit of detection [0.3 nmol/L (n = 6) vs approximately 0.1 nmol/L for isotopic assay]. None of 34 negative sera tested (16 healthy controls, 10 patients with nonmyasthenia-related disease, 8 patients seronegative for MG) gave a value >0.3 nmol/L. Of the 35 positive myasthenic sera (with antibody values, previously determined by isotopic assay, of 0.4-1290 nmol/L) compared in the two assays, 32 tested positive with the Eu(3+) assay. Linear regression analysis yielded the equation: y = 1.035x - 0.013 nmol/L; S(y:x) = 0.172 nmol/L; r(2) = 0.977. CONCLUSIONS: The new time resolved fluorescence method for quantification of antibodies to nAChRs in MG patients provides a performance similar to that of the widely used isotopic assay and could be used in laboratories with restricted use of isotopes. PMID- 11861447 TI - Age-related reference values for serum selenium concentrations in infants and children. AB - BACKGROUND: Children are at particular risk for selenium deficiency, which has potentially serious medical implications. Reliable age-specific reference values for serum selenium concentrations in children are sparse, but are essential for the identification of selenium deficiency and decisions regarding selenium supplementation. METHODS: Using electrothermal atomic absorption spectrometry, we analyzed serum selenium concentrations from 1010 apparently healthy children (age range, 1 day to 18 years) and from 60 patients on a protein-restricted diet because of inborn errors of metabolism. Reference intervals were defined according to recommended guidelines. RESULTS: Medians for serum selenium concentrations showed a statistically significant age dependency: a decrease from the age <1 month (0.64 micromol/L) to 4 months (0.44 micromol/L); an increase to 0.62 micromol/L in the 4-12 months age group; constant values in children between 1 and 5 years of age (0.90 micromol/L); and an additional slight increase to reach a plateau between 5 and 18 years (0.99 micromol/L). Of 43 children older than 1 year and on a protein-restricted diet, 87% showed serum selenium concentrations below the 2.5 percentile. CONCLUSIONS: Because of nutritional changes, serum selenium concentrations are significantly higher in older children than in infants under 1 year of age. The application of age-adjusted reference values may provide more specific criteria for selenium supplementation. Long-term protein restriction in children is reflected by a failure to achieve higher serum selenium concentrations with increasing age. PMID- 11861448 TI - Quantification of holo-transcobalamin, a marker of vitamin B12 deficiency. PMID- 11861449 TI - Single-strand conformation polymorphism analysis for alcohol dehydrogenase 2 (ADH2) genotyping using nail clippings. PMID- 11861450 TI - Testing for porphobilinogen in urine. PMID- 11861451 TI - Preanalytical factors in the measurement of intact parathyroid hormone with the DPC IMMULITE assay. PMID- 11861452 TI - Minimizing error in the determination of P50. PMID- 11861453 TI - Optimization of nitric oxide chemiluminescence operating conditions for measurement of plasma nitrite and nitrate. PMID- 11861454 TI - Interchangeability of estimates of day-to-day imprecision between commercial control materials and serum pools. PMID- 11861455 TI - Lack of seasonal variation in C-reactive protein. PMID- 11861456 TI - Serum vitamin E and lipid-adjusted vitamin E assessment in Friedreich ataxia phenotype patients and unaffected family members. PMID- 11861457 TI - More rapid method for simultaneous measurement of tryptophan and kynurenine by HPLC. PMID- 11861458 TI - Methemoglobin interferes with the HemoCue B-Glucose Analyzer. PMID- 11861459 TI - Limitations of steroid determination by direct immunoassay. PMID- 11861460 TI - The tortuous road to the adoption of katal for the expression of catalytic activity by the General Conference on Weights and Measures. AB - BACKGROUND: The "unit" for "enzymic activity" (U = 1 micromol/min) was recommended by the International Union of Biochemistry and Molecular Biology (IUB) in 1961 and is widely used in medical laboratory reports. The general trend in metrology, however, is toward global standardization through defining units coherent with the International System of Units (SI). APPROACH: Several proposals were advanced from the IFCC, International Union of Pure and Applied Chemistry, and IUB regarding the definition for enzymic activity as well as the terms for kind-of-quantity, units, symbol, and dimension. In 1977, international agreement was reached between these bodies and WHO that "catalytic activity" (z), of a catalyst in a given system is defined by the rate of conversion in a measuring system (in mol/s) and expressed in "katal" (symbol, kat; equal to 1 mol/s). The katal is invariant of the measurement procedure, but the numerical quantity value is not. Gaining support for the katal from the final arbiter, the General Conference on Weights and Measures, was slow, but Resolution 12 of 1999 adopted the katal (symbol, kat) as a special name and symbol for the SI-derived unit, mol/s, used in measuring catalytic activity. CONCLUSIONS: Laboratory results for amounts of catalysts, including enzymes, measured by their catalytic activity can now officially be expressed in katals and are traceable to the SI provided that the specified indicator reaction reflects first-order kinetics. The conversion from "unit" is: 1 U = 16.667 x 10(-9) kat. Further derived quantities have coherent units such as kat/L, kat/kg, and kat/kat = 1. PMID- 11861461 TI - C-reactive protein concentrations in cerebral spinal fluid in gram-positive and gram-negative bacterial meningitis. PMID- 11861462 TI - Use of sodium measurement as a surrogate volume measurement in unmetered-blood collection devices. PMID- 11861466 TI - Patterning of the embryo: the first spatial decisions in the life of a mouse. AB - Although in most species the polarity of the embryo takes its roots from the spatial patterning of the egg, mammals were viewed as an exception. This was because the anteroposterior polarity of the mouse embryo could not be seen until gastrulation, and no developmental cues were known that could define polarity at earlier stages. Why should we now re-consider this view? While mechanisms of axis formation in mammals could, in principle, be unique, the evolutionary conservation of numerous other developmental processes raises the question of why mammals would have evolved a different way or timing of organising their embryonic polarity. Indeed, recent evidence shows that well before the onset of gastrulation, the mouse embryo initiates asymmetric patterns of gene expression in its visceral endoderm. Although this extra-embryonic tissue does not contribute to the body itself, it is involved in axis formation. Other recent work has revealed that spatial distribution of cells in the visceral endoderm can be traced back to polarity present at the blastocyst stage. These insights have raised the possibility that embryonic polarity might also originate early during development of mammalian embryos. Indeed it now appears that there are at least two spatial cues that operate in the mouse egg to shape polarity of the blastocyst. One of these is at the animal pole, which is defined by the site of female meiosis, and another is associated with the position of sperm entry. In this review I discuss these recent findings, which have led to the recognition that mouse embryos initiate development of their polarity at the earliest stages of their life. This novel perspective raises questions about the nature of cellular and molecular mechanisms that could convert developmental cues in the zygote to axes of the blastocyst, and hence into polarity of the post implantation embryo. It also brings to light the need to understand how such mechanisms could enable early mouse development to be so regulative. PMID- 11861467 TI - Math3 and NeuroD regulate amacrine cell fate specification in the retina. AB - The basic helix-loop-helix genes Math3 and NeuroD are expressed by differentiating amacrine cells, retinal interneurons. Previous studies have demonstrated that a normal number of amacrine cells is generated in mice lacking either Math3 or NEUROD: We have found that, in Math3-NeuroD double-mutant retina, amacrine cells are completely missing, while ganglion and Muller glial cells are increased in number. In the double-mutant retina, the cells that would normally differentiate into amacrine cells did not die but adopted the ganglion and glial cell fates. Misexpression studies using the developing retinal explant cultures showed that, although Math3 and NeuroD alone only promoted rod genesis, they significantly increased the population of amacrine cells when the homeobox gene Pax6 or Six3 was co-expressed. These results indicate that Math3 and NeuroD are essential, but not sufficient, for amacrine cell genesis, and that co-expression of the basic helix-loop-helix and homeobox genes is required for specification of the correct neuronal subtype. PMID- 11861468 TI - Rasp, a putative transmembrane acyltransferase, is required for Hedgehog signaling. AB - Members of the Hedgehog (Hh) family encode secreted molecules that act as potent organizers during vertebrate and invertebrate development. Post-translational modification regulates both the range and efficacy of Hh protein. One such modification is the acylation of the N-terminal cysteine of Hh. In a screen for zygotic lethal mutations associated with maternal effects, we have identified rasp, a novel Drosophila segment polarity gene. Analysis of the rasp mutant phenotype, in both the embryo and wing imaginal disc demonstrates that rasp does not disrupt Wnt/Wingless signaling but is specifically required for Hh signaling. The requirement of rasp is restricted only to those cells that produce Hh; hh transcription, protein levels and distribution are not affected by the loss of rasp. Molecular analysis reveals that rasp encodes a multipass transmembrane protein that has homology to a family of membrane bound O-acyl transferases. Our results suggest that Rasp-dependent acylation is necessary to generate a fully active Hh protein. PMID- 11861469 TI - The C. elegans even-skipped homologue, vab-7, specifies DB motoneurone identity and axon trajectory. AB - Locomotory activity is defined by the specification of motoneurone subtypes. In the nematode, C. elegans, DA and DB motoneurones innervate dorsal muscles and function to induce movement in the backwards or forwards direction, respectively. These two neurone classes express separate sets of genes and extend axons with oppositely directed trajectories; anterior (DA) versus posterior (DB). The DA specific homeoprotein UNC-4 interacts with UNC-37/Groucho to repress the DB gene, acr-5 (nicotinic acetylcholine receptor subunit). We show that the C. elegans even-skipped-like homoedomain protein, VAB-7, coordinately regulates different aspects of the DB motoneurone fate, in part by repressing unc-4. Wild-type DB motoneurones express VAB-7, have posteriorly directed axons, express ACR-5 and lack expression of the homeodomain protein UNC-4. In a vab-7 mutant, ectopic UNC 4 represses acr-5 and induces an anteriorly directed DB axon trajectory. Thus, vab-7 indirectly promotes DB-specific gene expression and posteriorly directed axon outgrowth by preventing UNC-4 repression of DB differentiation. Ectopic expression of VAB-7 also induces DB traits in an unc-4-independent manner, suggesting that VAB-7 can act through a parallel pathway. This work supports a model in which a complementary pair of homeodomain transcription factors (VAB-7 and UNC-4) specifies differences between DA and DB neurones through inhibition of the alternative fates. The recent findings that Even-skipped transcriptional repressor activity specifies neurone identity and axon guidance in the mouse and Drosophila motoneurone circuit points to an ancient origin for homeoprotein dependent mechanisms of neuronal differentiation in the metazoan nerve cord. PMID- 11861471 TI - Characterization of the head organizer in hydra. AB - A central process in the maintenance of axial patterning in the adult hydra is the head activation gradient, i.e. the potential to form a secondary axis, which is maximal in the head and is graded down the body column. Earlier evidence suggested that this gradient was based on a single parameter. Using transplantation experiments, we provide evidence that the hypostome, the apical part of the head, has the characteristics of an organizer in that it has the capacity to induce host tissue to form most of the second axis. By contrast, tissue of the body column has a self-organizing capacity, but not an inductive capacity. That the inductive capacity is confined to the hypostome is supported by experiments involving a hypostome-contact graft. The hypostome, but not the body column, transmits a signal(s) leading to the formation of a second axis. In addition, variations of the transplantation grafts and hypostome-contact grafts provide evidence for several characteristics of the organizer. The inductive capacity of the head and the self-organizing capacity of the body column are based on different pathways. Head inhibition, yya signal produced in the head and transmitted to the body column to prevent head formation, represses the effect of the inducing signal by interfering with formation of the hypostome/organizer. These results indicate that the organizer characteristics of the hypostome of an adult hydra are similar to those of the organizer region of vertebrate embryos. They also indicate that the Gierer-Meinhardt model provides a reasonable framework for the mechanisms that underlie the organizer and its activities. In addition, the results suggest that a region of an embryo or adult with the characteristics of an organizer arose early in metazoan evolution. PMID- 11861470 TI - Bimodal functions of Notch-mediated signaling are involved in neural crest formation during avian ectoderm development. AB - Neural crest is induced at the junction of epidermal ectoderm and neural plate by the mutual interaction of these tissues. In previous studies, BMP4 has been shown to pattern the ectodermal tissues, and BMP4 can induce neural crest cells from the neural plate. In this study, we show that epidermally expressed Delta1, which encodes a Notch ligand, is required for the activation and/or maintenance of Bmp4 expression in this tissue, and is thus indirectly required for neural crest induction by BMP4 at the epidermis-neural plate boundary. Notch activation in the epidermis additionally inhibits neural crest formation in this tissue, so that neural crest generation by BMP4 is restricted to the junction. PMID- 11861472 TI - Low density detergent-insoluble membrane of Xenopus eggs: subcellular microdomain for tyrosine kinase signaling in fertilization. AB - Protein-tyrosine phosphorylation plays an important role in egg activation signaling at fertilization. We show that in Xenopus, fertilization stimulates a rapid and transient tyrosine phosphorylation of egg proteins, as revealed by immunoblotting with anti-phosphotyrosine antibody. Immunofluorescent microscopic analysis demonstrated that the phosphorylation occurs in cortical area of the egg animal hemisphere. To further characterize subcellular compartment for fertilization-dependent tyrosine kinase signaling, we isolated low density detergent-insoluble membrane (LD-DIM) fraction from Xenopus eggs. The egg LD-DIM was enriched in cholesterol and GM1 ganglioside. It also contained signaling molecules such as Xyk (Xenopus egg Src), Gq alpha, Ras, integrin beta 1 and CD9. Fertilization stimulated tyrosine phosphorylation of Xyk and some other LD-DIM proteins. Remarkably, sperm stimulated tyrosine phosphorylation of the LD-DIM proteins in vitro. The sperm-dependent phosphorylation was sensitive to the tyrosine kinase inhibitors PP2 and genistein. We found that pretreatment of eggs with methyl-beta-cyclodextrin, a cholesterol-binding substance, led to a decrease in cholesterol, Xyk and sperm-induced tyrosine phosphorylation in LD-DIM. In methyl-beta-cyclodextrin-treated eggs, sperm-induced Ca(2+) transient and first cell division were also inhibited. These findings suggest that the egg LD-DIM might serve as subcellular microdomain for tyrosine kinase signaling in Xenopus egg fertilization. PMID- 11861473 TI - The chicken beta-globin insulator element conveys chromatin boundary activity but not imprinting at the mouse Igf2/H19 domain. AB - Imprinting of the mouse insulin-like growth factor 2 (Igf2) and H19 genes is regulated by an imprinting control region (ICR). The hypomethylated maternal copy functions as a chromatin insulator through the binding of CTCF and prevents Igf2 activation in cis, while hypermethylation of the paternal copy inactivates insulator function and leads to inactivation of H19 in cis. The specificity of the ICR sequence for mediating imprinting and chromatin insulation was investigated by substituting it for two copies of the chicken beta-globin insulator element, (Ch beta GI)(2), in mice. This introduced sequence resembles the ICR in size, and in containing CTCF-binding sites and CpGs, but otherwise lacks homology. On maternal inheritance, the (Ch beta GI)(2) was hypomethylated and displayed full chromatin insulator activity. Monoallelic expression of Igf2 and H19 was retained and mice were of normal size. These results suggest that the ICR sequence, aside from CTCF-binding sites, is not uniquely specialized for chromatin insulation at the Igf2/H19 region. On paternal inheritance, the (Ch beta GI)(2) was also hypomethylated and displayed strong insulator activity- fetuses possessed very low levels of Igf2 RNA and were greatly reduced in size, being as small as Igf2-null mutants. Furthermore, the paternal H19 allele was active. These results suggest that differential ICR methylation in the female and male germ lines is not acquired through differential binding of CTCF. Rather, it is likely to be acquired through a separate or downstream process. PMID- 11861474 TI - The zebrafish spiel-ohne-grenzen (spg) gene encodes the POU domain protein Pou2 related to mammalian Oct4 and is essential for formation of the midbrain and hindbrain, and for pre-gastrula morphogenesis. AB - In early embryonic development, the brain is divided into three main regions along the anteroposterior axis: the forebrain, midbrain and hindbrain. Through retroviral insertional mutagenesis and chemical mutagenesis experiments in zebrafish, we have isolated mutations that cause abnormal hindbrain organization and a failure of the midbrain-hindbrain boundary (MHB) to form, a region that acts as an organizer for the adjacent brain regions. The mutations fail to complement the spiel-ohne-grenzen (spg) mutation, which causes a similar phenotype, but for which the affected gene is unknown. We show through genetic mapping, cloning of the proviral insertion site and allele sequencing that spg mutations disrupt pou2, a gene encoding the Pou2 transcription factor. Based on chromosomal synteny, phylogenetic sequence comparison, and expression and functional data, we suggest that pou2 is the zebrafish ortholog of mouse Oct3/Oct4 and human POU5F1. For the mammalian genes, a function in brain development has so far not been described. In the absence of functional pou2, expression of markers for the midbrain, MHB and the hindbrain primordium (pax2.1, wnt1, krox20) are severely reduced, correlating with the neuroectoderm-specific expression phase of pou2. Injection of pou2 mRNA restores these defects in spg mutant embryos, but does not activate these markers ectopically, demonstrating a permissive role for pou2. Injections of pou2-morpholinos phenocopy the spg phenotype at low concentration, further proving that spg encodes pou2. Two observations suggest that pou2 has an additional earlier function: higher pou2 morpholino concentrations specifically cause a pre-gastrula arrest of cell division and morphogenesis, and expression of pou2 mRNA itself is reduced in spg homozygous embryos at this stage. These experiments suggest two roles for pou2. Initially, Pou2 functions during early proliferation and morphogenesis of the blastomeres, similar to Oct3/4 in mammals during formation of the inner cell mass. During zebrafish brain formation, Pou2 then functions a second time to activate gene expression in the midbrain and hindbrain primordium, which is reflected at later stages in the specific lack in spg embryos of the MHB and associated defects in the mid- and hindbrain. PMID- 11861475 TI - Spiel-ohne-grenzen/pou2 mediates regional competence to respond to Fgf8 during zebrafish early neural development. AB - Neural patterning of the vertebrate brain starts within the ectoderm during gastrulation and requires the activity of organizer cell populations in the neurectoderm. One such organizer is located at the prospective midbrain-hindbrain boundary (MHB) and controls development of the midbrain and the anterior hindbrain via the secreted signaling molecule Fgf8. However, little is known about how the ability of neural precursors to respond to Fgf8 is regulated. We have studied the function of the zebrafish spiel-ohne-grenzen (spg) gene in early neural development. Genetic mapping and molecular characterization presented in the accompanying paper revealed that spg mutations disrupt the pou2 gene, which encodes a POU domain transcription factor that is specifically expressed in the MHB primordium, and is orthologous to mammalian Oct3/Oct4. We show that embryos homozygous for spg/pou2 have severe defects in development of the midbrain and hindbrain primordium. Key molecules that function in the formation of the MHB, such as pax2.1, spry4, wnt1, her5, eng2 and eng3, and in hindbrain development, such as krox20, gbx2, fkd3 and pou2, are all abnormal in spg mutant embryos. By contrast, regional definition of the future MHB in the neuroectoderm by complementary expression of otx2 and gbx1, before the establishment of the complex regulatory cascade at the MHB, is normal in spg embryos. Moreover, the Fgf8 and Wnt1 signaling pathways are activated normally at the MHB but become dependent on spg towards the end of gastrulation. Therefore, spg plays a crucial role both in establishing and in maintaining development of the MHB primordium. Transplantation chimeras show that normal spg function is required within the neuroectoderm but not the endomesoderm. Importantly, gain-of-function experiments by mRNA injection of fgf8 and pou2 or Fgf8 bead implantations, as well as analysis of spg-ace double mutants show that spg embryos are insensitive to Fgf8, although Fgf receptor expression and activity of the downstream MAP kinase signaling pathway appear intact. We suggest that spg/pou2 is a transcription factor that mediates regional competence to respond to Fgf8 signaling. PMID- 11861477 TI - Distinct functions of TBP and TLF/TRF2 during spermatogenesis: requirement of TLF for heterochromatic chromocenter formation in haploid round spermatids. AB - TLF (TBP-like factor) is a protein commonly thought to belong to the general transcription initiation complex. TLF is evolutionarily conserved and has been shown to be essential for early development in C. elegans, zebrafish and Xenopus. In mammals however, TLF has a specialised function, as revealed by targeted mutation of the gene in the mouse germline. The TLF mutation elicits a complete arrest of late spermiogenesis and increased haploid cell apoptosis. We explored in more detail the molecular function that TLF plays in the differentiation program of male germ cells. A comparison of TBP and TLF reveals drastic differences, both in their temporal expression pattern and in their intracellular location. While TBP is ubiquitously expressed, TLF expression is strictly developmentally regulated, being very high in late pachytene spermatocytes, suggesting a function prior to the apoptosis of the haploid cells. A refined study of TLF-deficient mice reveals defective acrosome formation in early stage spermatids. Most importantly, our results uncover an unsuspected function of TLF in chromatin organisation. Indeed, early spermatids in TLF-deficient mice display a fragmentation of the chromocenter, a condensed structure formed by the association of centromeric heterochromatin and containing the HP1 proteins. This defect is likely to be the primary cause of spermatogenic failure in the TLF mutant mice. PMID- 11861476 TI - A genetic hierarchy establishes mitogenic signalling and mitotic competence in the renal tubules of Drosophila. AB - Cell proliferation in the developing renal tubules of Drosophila is strikingly patterned, occurring in two phases to generate a consistent number of tubule cells. The later phase of cell division is promoted by EGF receptor signalling from a specialised subset of tubule cells, the tip cells, which express the protease Rhomboid and are thus able to secrete the EGF ligand, Spitz. We show that the response to EGF signalling, and in consequence cell division, is patterned by the specification of a second cell type in the tubules. These cells are primed to respond to EGF signalling by the transcription of two pathway effectors, PointedP2, which is phosphorylated on pathway activation, and Seven up. While expression of pointedP2 is induced by Wingless signalling, seven up is initiated in a subset of the PointedP2 cells through the activity of the proneural genes. We demonstrate that both signalling and responsive cells are set aside in each tubule primordium from a proneural gene-expressing cluster of cells, in a two-step process. First, a proneural cluster develops within the domain of Wingless-activated, pointedP2-expressing cells to initiate the co expression of seven up. Second, lateral inhibition, mediated by the neurogenic genes, acts within this cluster of cells to segregate the tip cell precursor, in which proneural gene expression strengthens to initiate rhomboid expression. As a consequence, when the precursor cell divides, both daughters secrete Spitz and become signalling cells. Establishing domains of cells competent to transduce the EGF signal and divide ensures a rapid and reliable response to mitogenic signalling in the tubules and also imposes a limit on the extent of cell division, thus preventing tubule hyperplasia. PMID- 11861478 TI - Manipulation of leaf shape by modulation of cell division. AB - The role of cell division as a causal element in plant morphogenesis is debatable, with accumulating evidence supporting the action of cell division independent mechanisms. To directly test the morphogenic function of cell division, we have utilised a microinduction technique to locally and transiently manipulate the expression in transgenic plants of two genes encoding putative effectors of the cell cycle, a tobacco A-type cyclin and a yeast cdc25. The results show that local expression of these genes leads to modulation of cell division patterns. Moreover, whereas altered cell division in the apical meristem had no influence on organogenesis, local induction of cell proliferation on the flanks of young leaf primordia led to a dramatic change in lamina development and, thus, leaf shape. These data indicate that the role of cell division in plant morphogenesis is context dependent and identify cell division in the leaf primordium as a potential target for factors regulating leaf shape. PMID- 11861479 TI - Mice that lack astrotactin have slowed neuronal migration. AB - The cortical regions of the brain are laminated as a result of directed migration of precursor cells along glia during development. Previously, we have used an assay system to identify astrotactin as a neuronal ligand for migration on glial fibers. To examine the function of astrotactin in vivo, we generated a null mutation by targeted gene disruption. The cerebella of astrotactin null mice are approximately 10% smaller than wild type. In vitro and in vivo cerebellar granule cell assays show a decrease in neuron-glial binding, a reduction in migration rates and abnormal development of Purkinje cells. Consequences of this are poorer balance and coordination. Thus, astrotactin functions in migration along glial processes in vivo, a process required for generating laminar structures and for the development of synaptic partner systems. PMID- 11861480 TI - Patterning of angiogenesis in the zebrafish embryo. AB - Little is known about how vascular patterns are generated in the embryo. The vasculature of the zebrafish trunk has an extremely regular pattern. One intersegmental vessel (ISV) sprouts from the aorta, runs between each pair of somites, and connects to the dorsal longitudinal anastomotic vessel (DLAV). We now define the cellular origins, migratory paths and cell fates that generate these metameric vessels of the trunk. Additionally, by a genetic screen we define one gene, out of bounds (obd), that constrains this angiogenic growth to a specific path. We have performed lineage analysis, using laser activation of a caged dye and mosaic construction to determine the origin of cells that constitute the ISV. Individual angioblasts destined for the ISVs arise from the lateral posterior mesoderm (LPM), and migrate to the dorsal aorta, from where they migrate between somites to their final position in the ISVs and dorsal longitudinal anastomotic vessel (DLAV). Cells of each ISV leave the aorta only between the ventral regions of two adjacent somites, and migrate dorsally to assume one of three ISV cell fates. Most dorsal is a T-shaped cell, based in the DLAV and branching ventrally; the second constitutes a connecting cell; and the third an inverted T-shaped cell, based in the aorta and branching dorsally. The ISV remains between somites during its ventral course, but changes to run mid somite dorsally. This suggests that the pattern of ISV growth ventrally and dorsally is guided by different cues. We have also performed an ENU mutagenesis screen of 750 mutagenized genomes and identified one mutation, obd that disrupts this pattern. In obd mutant embryos, ISVs sprout precociously at abnormal sites and migrate anomalously in the vicinity of ventral somite. The dorsal extent of the ISV is less perturbed. Precocious sprouting can be inhibited in a VEGF morphant, but the anomalous site of origin of obd ISVs remains. In mosaic embryos, obd somite causes adjacent wild-type endothelial cells to assume the anomalous ISV pattern of obd embryos. Thus, the launching position of the new sprout and its initial trajectory are directed by inhibitory signals from ventral somites. Zebrafish ISVs are a tractable system for defining the origins and fates of vessels, and for dissecting elements that govern patterns of vessel growth. PMID- 11861481 TI - The specification of noradrenergic locus coeruleus (LC) neurones depends on bone morphogenetic proteins (BMPs). AB - The role of BMPs in the development of the major noradrenergic centre of the brain, the locus coeruleus (LC), was investigated. LC generation is reflected by initial expression of the transcription factors Phox2a and Phox2b in dorsal rhombomere1 (r1), followed by expression of dopamine-beta-hydroxylase and tyrosine hydroxylase. Bmp5 is expressed in the dorsal neuroepithelium in proximity to Phox2-expressing cells. BMP inhibition in stage 10 chick embryos resulted in the lack of LC neurones or in their generation at the dorsal midline, and loss of roof plate and rhombic lip, but it did not affect neural crest development. These results reveal late essential BMP functions in the specification of dorsal neuronal phenotypes in r1, including LC neurones, and in the development of dorsal midline structures. PMID- 11861482 TI - EGFR signalling inhibits Capicua-dependent repression during specification of Drosophila wing veins. AB - Localised activation of the Ras/Raf pathway by Epidermal Growth Factor Receptor (EGFR) signalling specifies the formation of veins in the Drosophila wing. However, little is known about how the EGFR signal regulates transcriptional responses during the vein/intervein cell fate decision. We provide evidence that EGFR signalling induces expression of vein-specific genes by inhibiting the Capicua (Cic) HMG-box repressor, a known regulator of embryonic body patterning. Lack of Cic function causes ectopic expression of EGFR targets such as argos, ventral veinless and decapentaplegic and leads to formation of extra vein tissue. In vein cells, EGFR signalling downregulates Cic protein levels in the nucleus and relieves repression of vein-specific genes, whereas intervein cells maintain high levels of Cic throughout larval and pupal development, repressing the expression of vein-specific genes and allowing intervein differentiation. However, regulation of some EGFR targets such as rhomboid appears not to be under direct control of Cic, suggesting that EGFR signalling branches out in the nucleus and controls different targets via distinct mediator factors. Our results support the idea that localised inactivation of transcriptional repressors such as Cic is a rather general mechanism for regulation of target gene expression by the Ras/Raf pathway. PMID- 11861483 TI - A characterization of the effects of Dpp signaling on cell growth and proliferation in the Drosophila wing. AB - Cell proliferation and patterning must be coordinated for the development of properly proportioned organs. If the same molecules were to control both processes, such coordination would be ensured. Here we address this possibility in the Drosophila wing using the Dpp signaling pathway. Previous studies have shown that Dpp forms a gradient along the AP axis that patterns the wing, that Dpp receptors are autonomously required for wing cell proliferation, and that ectopic expression of either Dpp or an activated Dpp receptor, Tkv(Q253D), causes overgrowth. We extend these findings with a detailed analysis of the effects of Dpp signaling on wing cell growth and proliferation. Increasing Dpp signaling by expressing Tkv(Q253D) accelerated wing cell growth and cell cycle progression in a coordinate and cell-autonomous manner. Conversely, autonomously inhibiting Dpp signaling using a pathway specific inhibitor, Dad, or a mutation in tkv, slowed wing cell growth and division, also in a coordinate fashion. Stimulation of cell cycle progression by Tkv(Q253D) was blocked by the cell cycle inhibitor RBF, and required normal activity of the growth effector, PI3K. Among the known Dpp targets, vestigial was the only one tested that was required for Tkv(Q253D) induced growth. The growth response to altering Dpp signaling varied regionally and temporally in the wing disc, indicating that other patterned factors modify the response. PMID- 11861484 TI - Headless flies produced by mutations in the paralogous Pax6 genes eyeless and twin of eyeless. AB - The two Pax6 gene homologs eyeless and twin of eyeless play decisive early roles in Drosophila eye development. Strong mutants of twin of eyeless or of eyeless are headless, which suggests that they are required for the development of all structures derived from eye-antennal discs. The activity of these genes is crucial at the very beginning of eye-antennal development in the primordia of eye antennal discs when eyeless is first activated by the twin of eyeless gene product. This activation does not strictly depend on the Twin of eyeless protein, but is temperature-dependent in its absence. Twin of eyeless acts also in parallel to the eyeless gene and exerts functions that are partially redundant with those of Eyeless, while Eyeless is mainly required to prevent early cell death and promote eye development in eye-antennal discs. PMID- 11861485 TI - Hunchback is required for the specification of the early sublineage of neuroblast 7-3 in the Drosophila central nervous system. AB - The Drosophila ventral nerve cord (VNC) derives from neuroblasts (NBs), which mostly divide in a stem cell mode and give rise to defined NB lineages characterized by specific sets of sequentially generated neurons and/or glia cells. To understand how different cell types are generated within a NB lineage, we have focused on the NB7-3 lineage as a model system. This NB gives rise to four individually identifiable neurons and we show that these cells are generated from three different ganglion mother cells (GMCs). The finding that the transcription factor Hunchback (Hb) is expressed in the early sublineage of NB7 3, which consists of the early NB and the first GMC (GMC7-3a) and its progeny (EW1 and GW), prompted us to investigate its possible role in NB7-3 lineage development. Our analysis revealed that loss of hb results in a lack of the normally Hb-positive neurons, while the later-born neurons (designated as EW2 and EW3) are still present. However, overexpression of hb in the whole lineage leads to additional cells with the characteristics of GMC7-3a-derived neurons, at the cost of EW2 and EW3. Thus, hb is an important determinant in specifying early sublineage identity in the NB7-3 lineage. Using Even-skipped (Eve) as a marker, we have additionally shown that hb is also needed for the determination and/or differentiation of several other early-born neurons, indicating that this gene is an important player in sequential cell fate specification within the Drosophila CNS. PMID- 11861486 TI - Cross-repressive interactions of identity genes are essential for proper specification of cardiac and muscular fates in Drosophila. AB - In Drosophila embryos, founder cells that give rise to cardiac precursors and dorsal somatic muscles derive from dorsally located progenitors. Individual fates of founder cells are thought to be specified by combinatorial code of transcription factors encoded by identity genes. To date, a large number of identity genes have been identified; however, the mechanisms by which these genes contribute to cell fate specification remain largely unknown. We have analysed regulatory interactions of ladybird (lb), msh and even skipped (eve), the three identity genes specifying a subset of heart and/or dorsal muscle precursors. We show that deregulation of each of them alters the number of cells that express two other genes, thus changing the ratio between cardiac and muscular cells, and the ratio between different cell subsets within the heart and within the dorsal muscles. Specifically, we demonstrate that mutation of the muscle identity gene msh and misexpression of the heart identity gene lb lead to heart hyperplasia with similar cell fate modifications. In msh mutant embryos, the presumptive msh muscle cells switch on lb or eve expression and are recruited to form supernumerary heart or dorsal muscle cells, thus indicating that msh functions as a repressor of lb and eve. Similarly, overexpression of lb represses endogenous msh and eve activity, hence leading to the respecification of msh and eve positive progenitors, resulting in the overproduction of a subset of heart cells. As deduced from heart and muscle phenotypes of numb mutant embryos, the cell fate modifications induced by gain-of-function of identity genes are not lineage restricted. Consistent with all these observations, we propose that the major role of identity genes is to maintain their restricted expression by repressing other identity genes competent to respond positively to extrinsic signals. The cross-repressive interactions of identity genes are likely to ensure their localised expression over time, thus providing an essential element in establishing cell identity. PMID- 11861487 TI - Involvement of a proline-rich motif and RING-H2 finger of Deltex in the regulation of Notch signaling. AB - The Notch pathway is an evolutionarily conserved signaling mechanism that is essential for cell-cell interactions. The Drosophila deltex gene regulates Notch signaling in a positive manner, and its gene product physically interacts with the intracellular domain of Notch through its N-terminal domain. Deltex has two other domains that are presumably involved in protein-protein interactions: a proline-rich motif that binds to SH3-domains, and a RING-H2 finger motif. Using an overexpression assay, we have analyzed the functional involvement of these Deltex domains in Notch signaling. The N-terminal domain of Deltex that binds to the CDC10/Ankyrin repeats of the Notch intracellular domain was indispensable for the function of Deltex. A mutant form of Deltex that lacked the proline-rich motif behaved as a dominant-negative form. This dominant-negative Deltex inhibited Notch signaling upstream of an activated, nuclear form of Notch and downstream of full-length Notch, suggesting the dominant-negative Deltex might prevent the activation of the Notch receptor. We found that Deltex formed a homo multimer, and mutations in the RING-H2 finger domain abolished this oligomerization. The same mutations in the RING-H2 finger motif of Deltex disrupted the function of Deltex in vivo. However, when the same mutant was fused to a heterologous dimerization domain (Glutathione-S-Transferase), the chimeric protein had normal Deltex activity. Therefore, oligomerization mediated by the RING-H2 finger motif is an integral step in the signaling function of Deltex. PMID- 11861488 TI - Interactions between Hox-negative cephalic neural crest cells and the foregut endoderm in patterning the facial skeleton in the vertebrate head. AB - The vertebrate face contains bones that differentiate from mesenchymal cells of neural crest origin, which colonize the median nasofrontal bud and the first branchial arches. The patterning of individual facial bones and their relative positions occurs through mechanisms that remained elusive. During the early stages of head morphogenesis, an endodermal cul-de-sac, destined to become Sessel's pouch, underlies the nasofrontal bud. Reiterative outpocketings of the foregut then form the branchial pouches. We have tested the capacity of endoderm of the avian neurula to specify the facial skeleton by performing ablations or grafts of defined endodermal regions. Neural crest cells that do not express Hox genes respond to patterning cues produced regionally in the anterior endoderm to yield distinct skeletal components of the upper face and jaws. However, Hox expressing neural crest cells do not respond to these cues. Bone orientation is likewise dependent on the position of the endoderm relative to the embryonic axes. Our findings thus indicate that the endoderm instructs neural crest cells as to the size, shape and position of all the facial skeletal elements, whether they are cartilage or membrane bones. PMID- 11861489 TI - A mouse model of Alagille syndrome: Notch2 as a genetic modifier of Jag1 haploinsufficiency. AB - Alagille syndrome is a human autosomal dominant developmental disorder characterized by liver, heart, eye, skeletal, craniofacial and kidney abnormalities. Alagille syndrome is caused by mutations in the Jagged 1 (JAG1) gene, which encodes a ligand for Notch family receptors. The majority of JAG1 mutations seen in Alagille syndrome patients are null alleles, suggesting JAG1 haploinsufficiency as a primary cause of this disorder. Mice homozygous for a Jag1 null mutation die during embryogenesis and Jag1/+ heterozygous mice exhibit eye defects but do not exhibit other phenotypes characteristic of Alagille syndrome patients ( Xue, Y., Gao, X., Lindsell, C. E., Norton, C. R., Chang, B., Hicks, C., Gendron-Maguire, M., Rand, E. B., Weinmaster, G. and Gridley, T. (1999) HUM: Mol. Genet. 8, 723-730). Here we report that mice doubly heterozygous for the Jag1 null allele and a Notch2 hypomorphic allele exhibit developmental abnormalities characteristic of Alagille syndrome. Double heterozygous mice exhibit jaundice, growth retardation, impaired differentiation of intrahepatic bile ducts and defects in heart, eye and kidney development. The defects in bile duct epithelial cell differentiation and morphogenesis in the double heterozygous mice are similar to defects in epithelial morphogenesis of Notch pathway mutants in Drosophila, suggesting that a role for the Notch signaling pathway in regulating epithelial morphogenesis has been conserved between insects and mammals. This work also demonstrates that the Notch2 and Jag1 mutations interact to create a more representative mouse model of Alagille syndrome and provides a possible explanation of the variable phenotypic expression observed in Alagille syndrome patients. PMID- 11861490 TI - GnRH and GnRH receptor genes in the human genome. AB - Four different GnRHs and one GnRH receptor are reported to be expressed in various mammals, whereas 13 GnRHs and numerous GnRH receptors have been identified in various nonmammalian vertebrates. The nucleotide sequencing of the human genome provided the opportunity to determine which of these peptides and receptors might be expressed in primates. Of the four GnRHs reportedly expressed in mammals, only GnRH I (mammalian GnRH) and GnRH II (chicken GnRH II) genes were identified in the human genome. Three GnRH receptor or receptor-like genes were identified: 1) the well-established GnRH I receptor gene located on chromosome 4; 2) an apparent GnRH II receptor gene located on chromosome 1, and; 3) a sterile GnRH II receptor-like homolog gene on chromosome 14. A cDNA cloned from monkey RNA that was 96% identical with the putative human GnRH receptor type II gene encoded a 379-amino acid G protein-coupled/7-transmembrane receptor having a C terminal cytoplasmic tail. The experimentally expressed GnRH II receptor was functional with and specific for GnRH II, and, unlike the GnRH I receptor, desensitized to continuous GnRH treatment. GnRH II receptor mRNA is expressed ubiquitously in human tissues. Significant questions remain about the potential functions of the primate GnRH II receptor such as regulation of gonadotropin secretion, female sexual behavior, and tumor cell growth; also, about whether it is expressed as a full-length, functional gene transcript in humans. PMID- 11861491 TI - Dopamine-D2-mediated inhibition of TRH-induced PLC activation in pituitary cells direct or indirect? PMID- 11861492 TI - Evidence for a direct negative coupling between dopamine-D2 receptors and PLC by heterotrimeric Gi1/2 proteins in rat anterior pituitary cell membranes. AB - Dopamine (DA) is known to inhibit basal and hormone TRH- or angiotensin II (AngII)-stimulated PRL secretion and inositol phosphate accumulation in rat pituitary cells in primary culture. This inhibition persists when cells are incubated in a calcium-free medium (a condition in which DA could not inhibit PLC activities by blocking calcium influx) and is abolished by a Pertussis toxin treatment. These data suggest that DA receptor could be negatively coupled to PLC by a direct mechanism involving a Pertussis toxin-sensitive G protein. To demonstrate this hypothesis, we measured PLC activities on crude plasma membranes obtained from rat pituitary cells in primary culture grown in the presence of tritiated myo-inositol. We showed that 1) DA and quinpirole or RU24926 (specific D2 agonists) inhibited both basal and TRH- or AngII-stimulated membrane PLC activities. 2) Such inhibitions were completely prevented by sulpiride (specific D2 antagonist). 3) Heterotrimeric Gi1/2 proteins coupled the DA receptors to PLC because DA inhibitions were completely reversed by preincubation either with Pertussis toxin or with a specific G(alpha)i1/(alpha)i2 antibody. Such data are in favor of the existence of a direct negative coupling between DA-D2 receptor and PLC on a native physiological plasma membrane model. PMID- 11861493 TI - Cloning and distribution of galanin-like peptide mRNA in the hypothalamus and pituitary of the macaque. AB - Galanin-like peptide (GALP) is a newly discovered hypothalamic neuropeptide, which is regulated by leptin and implicated in the regulation of GnRH secretion in the rodent. We searched the human genome database and determined that the human GALP gene comprises six exons, as has been shown for human galanin. We used rapid amplification of cDNA ends to clone a full-length cDNA (802 bp) of the macaque homologue of GALP and found it to be highly conserved between human and macaque at both the nucleotide (93%) and peptide (94%) levels. Mature GALP is predicted to be 60 amino acids in the macaque as in other species, and the region of GALP (9-21) that shows homology to the N-terminal 13 amino acids of galanin is perfectly conserved. We mapped the distribution of GALP mRNA in the hypothalamus and pituitary of the macaque by in situ hybridization and observed that, as in rodent species, the expression of GALP mRNA is confined to the arcuate nucleus, median eminence, and neurohypophysis. Using double-label in situ hybridization, we found that nearly all (98%) GALP mRNA-expressing cells in the arcuate nucleus also express mRNA for the long form of the leptin receptor. These findings suggest that a leptin-GALP signaling pathway exists in a primate species. PMID- 11861495 TI - A novel in vivo rabbit model of hypercatabolic critical illness reveals a biphasic neuroendocrine stress response. AB - High doses of GH, used to induce anabolism in prolonged critically ill patients, unexpectedly increased mortality. To further explore underlying mechanisms, a valid animal model is needed. Such a model is presented in this study. Seven days after arterial and venous cannulae placement, male New Zealand White rabbits were randomly allocated to a control or a critically ill group. To induce prolonged critical illness, a template controlled 15% deep dermal burn injury was imposed under combined general and regional (paravertebral) anesthesia. Subsequently, critically ill rabbits received supplemental analgesia and were parenterally fed with glucose, insulin, amino acids, and lipids. On d 1 and d 8 after randomization, acute and chronic spontaneous hormonal profiles of GH, TSH, and PRL secretion were obtained by sampling blood every 15 min for 7 h. Furthermore, GH, TSH, and PRL responses to an iv bolus of GH-releasing peptide 2 (GHRP-2) + TRH were documented on d 0, 1, and 8. Hemodynamic status and biochemical parameters were evaluated on d 0, 1, 3, 5, and 8, after which animals were killed and relative wet weight and water content of organs was determined. Compared with controls, critically ill animals exhibited transient metabolic acidosis on d 1 and weight loss, organ wasting, systolic hypertension, and pronounced anemia on d 8. On d 1, pulsatile GH secretion doubled in the critically ill animals compared with controls, and decreased again on d 8 in the presence of low plasma IGF-I concentrations from d 1 to d 8. GH responses to GHRP-2 + TRH were elevated on d 1 and increased further on d 8 in the critically ill animals. Mean TSH concentrations were identical in both groups on d 1 and 8, in the face of dramatically suppressed plasma T(4) and T(3) concentrations in the critically ill animals. PRL secretion was impaired in the critically ill animals exclusively on d 8. TSH and PRL responses to GHRP-2 and TRH were increased only on d 1. In conclusion, this rabbit model of acute and prolonged critical illness reveals several of the clinical, biochemical, and endocrine manifestations of the human counterpart. PMID- 11861498 TI - Expression, hormonal regulation, and cyclic variation of chemokines in the rat ovary: key determinants of the intraovarian residence of representatives of the white blood cell series. AB - A growing body of evidence suggests that mammalian ovulation bears similarities to local inflammatory reactions. Monocytes/macrophages, eosinophils, and neutrophils are known to infiltrate the area surrounding the dominant follicle before ovulation. Candidate local chemoattractants may include a family of small cytokines, also known as chemokines. In the present study, quantitative RT-PCR was used to initially identify and quantify the chemokines expressed in the preovulatory rat ovary. The chemokines monocyte chemotatic protein 1 (MCP-1), MCP 3, macrophage inflammatory protein 1alpha (MIP-1alpha), MIP-1beta, MIP-1gamma, regulated upon activation normal T cell expressed and secreted, eotaxin, interferon-inducible protein of 10 kDa, growth-regulated oncogene, lymphotactin, and fractalkine were all expressed in the PMSG-primed rat ovary 6 h post human CG. C10, T cell activation gene 3, exodus, exodus-2, cytokine-induced neutrophil chemoattractant-2, MIP-2, and lipopolysaccharide-induced C-X-C were not expressed in the PMSG-primed rat ovary 6 h post human CG. The cyclic variation of the ovary positive chemokines was also evaluated throughout the course of a superovulated ovarian cycle. Significant preovulatory up-regulation relative to the untreated control state was documented for MCP-1 (18-fold), MCP-3 (12-fold), and growth regulated oncogene (25-fold). In contrast, the preovulatory ovarian expression of eotaxin, fractalkine and regulated upon activation normal T cell expressed and secreted was not increased. These observations suggest that intraovarian chemokines may be responsible for the cyclic intraovarian residence of representatives of the white blood cell series. PMID- 11861497 TI - Characterizaton of short isoforms of the leptin receptor in rat cerebral microvessels and of brain uptake of leptin in mouse models of obesity. AB - Leptin deficiency causes obesity in rodents and humans, but circulating levels of leptin are paradoxically elevated in obesity. The mechanisms underlying this leptin resistance are unknown, but may involve reduced leptin transport across the blood-brain barrier via short isoforms of the leptin receptor (Ob-R). Here, we first quantified short Ob-R mRNA expression in isolated rat cerebral microvessels constituting the blood-brain barrier and found that Ob-Ra and Ob-Rc mRNA were abundantly expressed in similar amounts. Second, brain uptake of leptin was reduced in mice lacking Ob-R. Third, brain uptake of leptin in New Zealand Obese mice, a strain that responds to central, but not peripheral, leptin, was reduced, suggesting that their obesity is at least partly due to deficient leptin transport into the brain. Fourth, brain uptake of leptin was significantly reduced in diet-induced obese mice. Neither New Zealand Obese mice nor diet induced obese mice exhibited significant decreases in Ob-R mRNA expression in isolated cerebral microvessels. These data support the ideas that short isoforms of Ob-R are involved in brain uptake of leptin and that impaired blood-brain barrier function contributes to the pathogenesis of obesity. However, the mechanisms by which obesity-related deficits in brain uptake of leptin occur remain to be defined. PMID- 11861499 TI - Impairment of liver GH receptor signaling by fasting. AB - Fasting causes a state of GH resistance responsible for low circulating IGF-I levels. To investigate whether this resistance may result from alterations in the GH signaling pathway, we determined the effects of fasting on the GH transduction pathway in rat liver. Forty-eight-hour fasted or fed male rats were injected with recombinant rat GH via the portal vein. Liver was removed 0 and 15 min after injection. Although GH stimulated Janus kinase 2 (JAK2) phosphorylation in all animals, this was severely blunted in fasted animals. Similarly, the phosphorylation of the GH receptor, although observed in both fasted and fed rats after GH injection, was markedly reduced in fasted rats. A rapid signal transducer and activator of transcription 5 (STAT5) tyrosine phosphorylation was also induced in the liver of fed animals in response to GH. In contrast, in fasted rats only a slight phosphorylated STAT5 signal was observed. The inhibitory effect of fasting on these GH signaling molecules occurred without changes in their protein content. Furthermore, the impairment of the JAK-STAT pathway in fasted animals was associated with increased liver suppressor of cytokine signaling 3 mRNA levels. Although glucocorticoids, which are increased by fasting, may cause GH resistance, adrenalectomy failed to prevent alterations in the JAK-STAT pathway caused by fasting. In conclusion, the GH resistance induced by fasting is associated with impairment of the JAK-STAT signaling pathway. This might contribute to the decrease in liver IGF-I production observed in fasting. PMID- 11861500 TI - Adrenal neutral cholesteryl ester hydrolase: identification, subcellular distribution, and sex differences. AB - Adrenals express a high level of neutral cholesteryl ester hydrolase (CEH) activity, and male rats have greater activity than females; however, the identity of the enzyme(s) responsible for this activity and the basis for the sex differences are unknown. Using mice in which hormone-sensitive lipase (HSL) was inactivated by homologous recombination (HSL -/-), neutral CEH activity was reduced more than 98% compared with controls. Female HSL -/- mice showed a reduction in stimulated corticosterone values. Mechanical separation of rat adrenals revealed less HSL in the outer than the inner cortex. Examination of subfractions of rat adrenals showed that immunoreactive HSL was prominently expressed in microsomes, with lesser amounts in the cytosol and little to no HSL in mitochondrial and nuclear fractions or the lipid droplet. Four- to 10-fold more neutral CEH activity was in the microsomal fraction than any other fraction. No sex differences in the expression or subcellular distribution of HSL protein were found; however, neutral CEH activity was lower in the microsomal fraction of females, and female adrenals contained more cholesteryl esters. Thus, HSL appears to be responsible for most, if not all, of adrenal neutral CEH activity, is prominently expressed in microsomes, and its activity is influenced by sex. PMID- 11861501 TI - CRH inhibits cell growth of human endometrial adenocarcinoma cells via CRH receptor 1-mediated activation of cAMP-PKA pathway. AB - CRH produced by human endometrial cells exerts decidualizing activity via an autocrine mechanism mediated via CRH-R1 receptors. We postulated that such activity exerted by CRH on normal endometrial cells might translate into an antiproliferative action on endometrial-derived malignancies, provided that neoplastic cells maintain the expression of CRH receptors. In this light, here we investigated the possible antiproliferative effects of CRH in an adenocarcinoma cell line derived from human endometrium. CRH induces time- and concentration dependent inhibition of Ishikawa cell growth, the maximal effect (50% inhibition) being achieved after 3 d of treatment with 10(-7) M CRH. A decrease in telomerase activity, which paralleled tumor growth inhibition, was also observed in CRH treated samples. The antiproliferative effect was confirmed by colony-formation assay for long-term survival. This effect was counteracted in a concentration dependent manner by both alpha-helical CRH and astressin; the former also showed intrinsic inhibitory activity. These findings suggested the involvement of CRH-R1 receptor subtype; this hypothesis was confirmed by RNase protection analysis showing the expression of human CRH-R1 mRNA. Experiments with the PKA inhibitor 14-22 amide and forskolin, as well as the measurement of intracellular cAMP, suggested the downstream involvement of cAMP-PKA pathway in CRH-induced inhibition of Ishikawa cell growth. PMID- 11861502 TI - Characterization of iodothyronine sulfatase activities in human and rat liver and placenta. AB - In conditions associated with high serum iodothyronine sulfate concentrations, e.g. during fetal development, desulfation of these conjugates may be important in the regulation of thyroid hormone homeostasis. However, little is known about which sulfatases are involved in this process. Therefore, we investigated the hydrolysis of iodothyronine sulfates by homogenates of V79 cells expressing the human arylsulfatases A (ARSA), B (ARSB), or C (ARSC; steroid sulfatase), as well as tissue fractions of human and rat liver and placenta. We found that only the microsomal fraction from liver and placenta hydrolyzed iodothyronine sulfates. Among the recombinant enzymes only the endoplasmic reticulum-associated ARSC showed activity toward iodothyronine sulfates; the soluble lysosomal ARSA and ARSB were inactive. Recombinant ARSC as well as human placenta microsomes hydrolyzed iodothyronine sulfates with a substrate preference for 3,3' diiodothyronine sulfate (3,3'-T(2)S) approximately T(3) sulfate (T(3)S) >> rT(3)S approximately T(4)S, whereas human and rat liver microsomes showed a preference for 3,3'-T(2)S > T(3)S >> rT(3)S approximately T(4)S. ARSC and the tissue microsomal sulfatases were all characterized by high apparent K(m) values (>50 microM) for 3,3'-T(2)S and T(3)S. Iodothyronine sulfatase activity determined using 3,3'-T(2)S as a substrate was much higher in human liver microsomes than in human placenta microsomes, although ARSC is expressed at higher levels in human placenta than in human liver. The ratio of estrone sulfate to T(2)S hydrolysis in human liver microsomes (0.2) differed largely from that in ARSC homogenate (80) and human placenta microsomes (150). These results suggest that ARSC accounts for the relatively low iodothyronine sulfatase activity of human placenta, and that additional arylsulfatase(s) contributes to the high iodothyronine sulfatase activity in human liver. Further research is needed to identify these iodothyronine sulfatases, and to study the physiological importance of the reversible sulfation of iodothyronines in thyroid hormone metabolism. PMID- 11861503 TI - Akt1 and Akt2 differently regulate muscle creatine kinase and myogenin gene transcription in insulin-induced differentiation of C2C12 myoblasts. AB - Insulin and IGFs are potent inducers of skeletal muscle differentiation. Although PI3K is known to be involved in skeletal muscle differentiation, its downstream targets in this process are not clearly defined. We investigated the roles of Akt and mammalian target of rapamycin (mTOR) in skeletal muscle differentiation. LY294002, a pharmacological inhibitor of PI3K, and the immunosuppressant rapamycin inhibited insulin-induced differentiation of C2C12 myoblasts. LY294002 and rapamycin suppressed myosin heavy chain expression and myotube formation. Transient reporter assays showed that both inhibitors repress muscle creatine kinase (MCK) and myogenin gene transcription. Heterologous expression of Akt1/PKB(alpha) potently suppressed MCK gene transcription without affecting myogenin gene transcription, whereas heterologous expression of Akt2 increased myogenin and MCK gene transcription. Finally, overexpression of myogenin rescued the inhibitory effect of rapamycin on MCK gene transcription, whereas it failed to rescue the inhibitory effect of LY294002 and Akt1. These results suggest that insulin regulates myogenic differentiation chiefly at the level of myogenin gene transcription via PI3K and mTOR. PI3K activity, but not mTOR, may regulate transcriptional activity of myogenin. Our data also suggest that Akt1 and Akt2 play distinct roles in myogenic differentiation. PMID- 11861504 TI - Gonadotropins, via cAMP, negatively regulate GATA-1 gene expression in testicular cells. AB - We and others demonstrated that the mRNAs encoding GATA-binding proteins, GATA-1 and GATA-4, were detected in mouse and rat testis, and in isolated rat Sertoli cells and testicular tumor cell lines derived from Leydig and Sertoli cells. In this study, we investigated the possible effects of gonadotropins and cAMP on the expression of GATA-binding protein genes in testicular cells. Unexpectedly, FSH negatively regulated GATA-1 (but not GATA-4) mRNA in a dose-dependent manner in primary cultures of rat Sertoli cells isolated from 21-d-old animals. GATA-1 mRNA was also negatively regulated by cAMP in a dose- and time-dependent manner in MA 10, a mouse Leydig tumor cell line. When 0.3 mM cAMP was administered to MA-10 cell cultures for 4 h, more than 95% of the GATA-1 mRNA and protein was abolished. The reduction of GATA-1 mRNA by cAMP can be mimicked by treatment with forskolin, which elevates intracellular cAMP levels. The inhibitory effect of cAMP was specific to the GATA-1 gene, given that GATA-4 and alpha-tubulin mRNA levels were not changed by any of the cAMP treatments. Inhibin alpha-subunit mRNA, on the other hand, was evidently increased by cAMP treatment in both MA-10 and Sertoli cells. However, inhibin alpha-subunit mRNA levels were elevated at 60 90 min before the suppression of GATA-1 mRNA detected. The inhibitory effect of cAMP on GATA-1 mRNA and protein was shown to be specific to testicular cells. The GATA-1 mRNA expressed in MEL, a mouse erythroid leukemia cell line, was not affected by cAMP. The reduction of GATA-1 mRNA by cAMP can be prevented when a translational inhibitor, cycloheximide, is added. In summary, we demonstrated that gonadotropins via cAMP negatively regulate the mRNA and protein levels of GATA-1, but not GATA-4, in testicular cells. The inhibitory effect on GATA-1 gene expression was specific to testicular cells and was not observed in erythroid cells. PMID- 11861505 TI - Aurintricarboxylic acid induces a distinct activation of the IGF-I receptor signaling within MDA-231 cells. AB - Aurintricarboxylic acid (ATA), a polymeric carboxylated triphenylmethane derivate, prevents apoptotic death in a variety of cell systems. Recently, we have shown that the survival promoting effect of ATA is transduced via activation of the IGF-I receptor (IGF-IR) signaling pathway. In breast cancer MDA-231 cells exposed either to the protein synthesis inhibitors cycloheximide or ricin or to the anticancer drug adriamycin, we have found that ATA, but not IGF-1, is a powerful antiapoptotic agent. The purpose of this study was to compare the ability of ATA and IGF-I to activate the IGF-IR signaling cascade and to correlate this ability to their survival potency. MDA-231 cells were exposed to ATA or IGF-I, up to 7 h, and the dynamics of activation of the IGF-IR signaling cascade was evaluated. Our results show that: 1) The amount of tyrosine phosphorylated IGF-IR proteins was greater after exposure to ATA, compared with IGF-I. 2) Two phosphorylated IGF-IR beta-subunits (a 95-kDa and a 75-kDa) were induced after exposure to ATA, whereas IGF-1 induced only the 95-kDa form. Immunoprecipitation of both receptor forms by antibodies against the alpha subunit and against the carboxy terminus of the beta-subunit of the IGF-IR suggests that the 75-kDa form could be the beta-chain truncated at the amino terminus above the alpha-beta disulphide bridges. 3) The ATA-activated IGF-IR forms underwent slow dephosphorylation, compared with a rapid dephosphorylation of the IGF-I activated receptor. 4) The insulin receptor substrate-1/2-associated PI3K, Shc proteins, and the kinases Akt and Erk1/2, downstream mediators of the antiapoptotic signaling by IGF-IR, were activated to a higher extent and for a longer time period by ATA, compared with IGF-I. Taken together, the sustained activation of the IGF-IR signaling pathway by ATA may explain its stronger antiapoptotic effect. We suggest that this enhanced activity, and the different susceptibility of the IGF-IR to certain proteases and phosphatases, may indicate a distinct conformation of the ATA-activated IGF-IR. PMID- 11861506 TI - Inhibition of cyclooxygenase-2 by natriuretic peptides. AB - The atrial natriuretic peptide (ANP) has been suggested to possess immunomodulatory potential because of its property to alter macrophage functions via its guanylate-cylcase- coupled A-receptor (NPR-A), such as inhibiting the expression of inducible nitric oxide synthase or TNF-alpha. The aim of this study was to investigate whether ANP influences COX-2. COX-2 expression in murine macrophages and in mice was induced by lipopolysaccharide. Release of PGE(2) and thromboxane B(2) was significantly reduced in the presence of ANP. C-type natriuretic peptide (CNP) also significantly reduced PGE(2)-accumulation in macrophages. Northern and Western blots showed that ANP attenuates COX-2 mRNA and protein. Reduction of neither COX-2 nor of PGE(2) production was significantly abrogated by an NPR-A antagonist, suggesting a pathway independent of cGMP. Furthermore, dibutyryl-cGMP did not affect PGE(2)-accumulation. cANF, the specific ligand for the natriuretic peptide (NP) clearance-receptor (NPR-C), significantly inhibited PGE(2)-production. Because some biological activities of ANP have been reported to be mediated via an NPR-C-mediated inhibition of adenylate-cyclase, we determined cAMP levels. ANP, CNP, and cANF significantly attenuated intracellular cAMP. In summary, ANP was shown to attenuate PGE(2) production of lipopolysaccharide-activated macrophages predominantly via the NP clearance-receptor. ANP reduces COX-2-protein and -mRNA levels. The inhibition seems to be mediated via NPR-C and related to an attenuation of cAMP production. PMID- 11861507 TI - The agonist activity of tamoxifen is inhibited by the short heterodimer partner orphan nuclear receptor in human endometrial cancer cells. AB - Short heterodimer partner (SHP) is an orphan nuclear receptor that interacts with ER(alpha) and ERbeta and inhibits E2-induced transcription. We examined how SHP affects tamoxifen's estrogen agonist activity in endometrial cells. We report that SHP interacts with 4-hydroxytamoxifen (4-OHT) or E2-occupied ER(alpha) in a temperature-dependent manner in vitro. In transient transfection assays, SHP inhibited 4-OHT-stimulated reporter gene activity from an estrogen response element (ERE) in ER-positive RL95-2 but not in HEC-1A human endometrial carcinoma cells transfected with ER(alpha) or ERbeta. SHP inhibited E2-induced transcriptional activity in ER(alpha)- or ERbeta-transfected HEC-1A or Chinese hamster ovary-K1 cells. SHP inhibition of E2 activity was greater for ER(alpha) than ERbeta from the nonpalindromic ERE in the pS2 gene promoter in Chinese hamster ovary-K1 but not HEC-1A cells. Thus, ER subtype, cell type, and ERE sequence influence SHP repressor activity. An ER(alpha) mutant lacking activator function-1 showed reduced inhibition by SHP. In glutathione S-transferase pull down experiments, SHP inhibited ER(alpha) dimerization, providing a possible mechanism to account for the inhibitory effect of SHP on ER activity. These results identify SHP as novel target for blocking 4-OHT agonist activity in endometrial cells. PMID- 11861508 TI - Overexpression of human stanniocalcin affects growth and reproduction in transgenic mice. AB - In mammals stanniocalcin (STC) is widely expressed, and in the kidney and gut it regulates serum calcium levels by promoting phosphate reabsorption. To shed further light on its functional significance in mammals we have created several lines of mice that express a human STC (hSTC) transgene. Three lines expressed the hSTC transgene, but only two lines exhibited high expression and contained circulating hSTC, and in these animals there was a reduction in postnatal growth (30-50%) that persisted after weaning. Moreover, even wild-type pups exhibited a growth retardation phenotype when nursed by a transgenic foster mother, and this implies that hSTC overexpression deleteriously affects maternal behavior and/or lactation. The reproductive potential of female transgenic mice was also compromised, as evidenced by significantly smaller litter sizes, but transgenic male fertility was unchanged even though the transgene was most highly expressed in testes. Interestingly, transgene-derived serum hSTC increased significantly after puberty and was severalfold higher in females than in males, suggesting a gender-specific mechanism for maintaining elevated circulating levels of STC. Blood analysis revealed that both transgenic lines had elevated phosphate and decreased alkaline phosphatase levels, indicative of altered kidney and bone metabolism. These studies provide the first evidence that STC is involved in growth and reproduction and reaffirm its role in mineral homeostasis. PMID- 11861509 TI - Requirement for ERK1/2 activation in the regulation of progesterone production in human granulosa-lutein cells is stimulus specific. AB - This study was conducted to determine whether the ERK1/2 family of MAPKs can be modulated by physiological regulators of the human corpus luteum, and whether this activation is important for progesterone secretion in human granulosa-lutein (hGL) cells. Human LH (hLH), hCG, and agents that indirectly elevate cAMP [cholera toxin, forskolin, (Bu)(2)cAMP], time- and dose-dependently activated ERK1/2 in hGL cells. ERK1/2 activation was reduced by preincubation with PKA inhibitors, including myristoylated PKI, suggesting that cAMP mediates ERK1/2 activation. Two structurally distinct inhibitors of MAPK kinase (MEK), PD 98059 and U 0126, abrogated hLH/hCG-induced ERK1/2 activation, but had no effect on hLH , hCG-, or 22R-hydroxycholesterol-stimulated progesterone secretion. In contrast, both inhibitors blocked cholera toxin-, forskolin-, and (Bu)(2)cAMP-induced ERK1/2 phosphorylation concomitant with a reduction in progesterone secretion. The known luteotropin, PGE(2), promoted MEK- and cAMP-dependent activation of ERK1/2, and inhibitors of either MEK or PKA decreased PGE(2)-induced progesterone synthesis. Our findings demonstrate that the requirement for ERK1/2 activation as a regulator of progesterone synthesis in hGL cells is stimulus dependent, and that the MEK inhibitor-sensitive step is distal to cAMP generation, but proximal to the conversion of cholesterol to pregnenolone. PMID- 11861510 TI - Somatostatin stimulates GH secretion in two porcine somatotrope subpopulations through a cAMP-dependent pathway. AB - Somatostatin (SRIF) inhibits GH release from rat somatotropes by reducing adenylate cyclase (AC) activity and the free cytosolic calcium concentration ([Ca(2+)](i)). In contrast, we have reported that SRIF can stimulate GH release in vitro from pig somatotropes. Specifically, 10(-7) and 10(-15) M SRIF stimulate GH release from a subpopulation of high density (HD) somatotropes isolated by Percoll gradient centrifugation, whereas in low density (LD) somatotropes only 10(-15) M SRIF induces such an effect. To ascertain the signaling pathways underlying this phenomenon, we assessed SRIF effects on second messengers in cultured LD and HD cells by measuring cAMP, IP turnover, and [Ca(2+)](i). Likewise, contribution of the corresponding signaling pathways to SRIF-induced GH release was evaluated by blocking AC, PLC, extracellular Ca(2+) influx, or intracellular Ca(2+) mobilization. Both 10(-7) and 10(-15) M SRIF increased cAMP, IP turnover, and [Ca(2+)](i) in HD cells. Conversely, in LD cells 10(-7) M SRIF reduced [Ca(2+)](i), but did not alter cAMP or IP, and 10(-15) M SRIF was without effect. Interestingly, SRIF-stimulated GH release was abolished in both subpopulations by AC blockade, but not by PLC inhibition. Furthermore, SRIF induced GH release was not reduced by blockade of extracellular Ca(2+) influx through voltage-sensitive channels or by depletion of thapsigargin-sensitive intracellular Ca(2+) stores. Therefore, SRIF stimulates GH secretion from cultured porcine somatotrope subpopulations through an AC/cAMP pathway-dependent mechanism that is seemingly independent of net increases in IP turnover or [Ca(2+)](i). These novel actions challenge classic views of SRIF as a mere inhibitor for somatotropes and suggest that it may exert a more complex, dual function in the control of porcine GH release, wherein molecular heterogeneity of somatotropes would play a critical role. PMID- 11861511 TI - Regulated expression of Wnts and Frizzleds at specific stages of follicular development in the rodent ovary. AB - Wnt ligands and Frizzled (Fz) G protein-coupled receptors impact cell fate, including embryonic development of the ovary. Because the role of these regulatory molecules during follicular development in the adult is not known, an RT-PCR survey was done. Wnt-4, Fz-4, and Fz-1 were among the transcripts detected, and each exhibited a specific pattern of expression. Fz-1 mRNA was low in preovulatory follicles of PMSG-treated mice but was increased within 4-12 h after an ovulatory surge of human CG. By in situ analysis, Fz-1 transcripts increased first in the theca cells and then in the granulosa cells of ovulating follicles but were low in corpora lutea. In contrast, Wnt-4, a critical factor in early ovarian development, was expressed in small preantral follicles. In addition, Wnt-4 was detected in preovulatory follicles and exhibited high levels in corpora lutea. A potential receptor for Wnt-4 in corpora lutea is Fz-4 that was also elevated in this tissue. Although Wnt-4 has been shown to function downstream of the PR in other tissues, Wnt-4 was not altered in follicles of PR null mice that fail to ovulate. Rather expression of Fz-1 was lower in ovaries of PR knockout mice, compared with normal littermates. Thus, specific Wnt/Fz are expressed at distinct stages of follicular development, suggesting multiple functions for this signaling pathway in the ovary. PMID- 11861512 TI - Murine relaxin-like factor promoter: functional characterization and regulation by transcription factors steroidogenic factor 1 and DAX-1. AB - The gene for mouse relaxin-like factor (RLF), a member of the insulin/IGF/relaxin family of hormones, appears to be predominantly expressed in testicular Leydig cells. Mice deficient in RLF have revealed a role for this peptide in testicular descent, but the regulatory mechanisms of its function are still insufficiently characterized. In the present study we showed that the RLF promoter was active in both mLTC-1 Leydig cells and luteinized KK-1 granulosa tumor cells. Interestingly, the activity of the RLF promoter as well as the expression of endogenous RLF correlated with the amount of steroidogenic factor 1 (SF-1) expression in the four cell lines tested. The highest transcriptional activity (29-fold over promoterless plasmid) was detected in mLTC-1 using the 188-bp promoter fragment immediately 5' of the CAP site, containing three consensus sequences for SF-1 binding. However, the promoter fragments including the 188-bp promoter also showed significant SF-1-independent promoter activity in both mLTC 1 and KK-1 cells, 8-fold induced over the promoterless construct. Mutagenesis studies showed that all three SF-1-binding sites were needed to obtain maximal SF 1-dependent trans-activation. The most distal SF-1-binding site at position -144 to -136 showed the highest affinity toward SF-1, but the promoter fragments, including the SF-1-binding site at position -115 to -107, showed the strongest response to SF-1 in terms of transcriptional activation. Moreover, DAX-1 inhibited RLF promoter activity in mLTC-1 Leydig tumor cells and totally abolished SF-1-dependent RLF expression in nonsteroidogenic HEK-293 cells. DAX-1 especially inhibited binding of SF-1 to the binding motifs locating at positions 64 to -56 and -115 to -107, whereas no decrease was seen in the expression of SF 1. Taken together, these observations suggest that the 188-bp RLF promoter includes elements for both SF-1-dependent and -independent gene expression in steroidogenic cells. The data, furthermore, indicate differential binding affinities for the three SF-1 binding motifs toward SF-1, of which the motif locating at position -115 to -107 was the most critical for the promoter activity. PMID- 11861513 TI - RhoA/Rho-kinase cascade is involved in oxytocin-induced rat uterine contraction. AB - The RhoA/Rho-kinase cascade is involved in various cellular functions, including migration, proliferation, and smooth muscle contraction. We examined the potential role of this pathway in oxytocin-induced uterine contraction. The specific Rho-kinase inhibitor Y-27632 inhibited oxytocin-induced rat uterine contraction on d 21 of pregnancy in a concentration-dependent manner, whereas the extent of this inhibition was reduced in the nonpregnant uterus. Y-27632 had no effect on oxytocin-induced intracellular Ca(2+) mobilization in myometrial cells. Immunoblot analysis showed that oxytocin increased the level of myosin light chain phosphorylation, and this increase was attenuated by Y-27632. Oxytocin increased the phosphorylation of myosin-binding subunit of myosin phosphatase, one of the major substrates of Rho-kinase, and this increase was reduced by Y 27632. The expression of Rho-kinase protein was shown to increase in the uterus during pregnancy compared with the nonpregnant uterus, whereas the expression of RhoA protein remained at the same level during pregnancy. RT-PCR and Northern blot analysis showed that the expression of Rho-kinase was up-regulated at the transcriptional level during pregnancy. These results suggest that the RhoA/Rho kinase pathway may have an important role in oxytocin-induced uterine contraction, and that up-regulation of Rho-kinase is involved in the mechanism underlying the increased contractility of the pregnant myometrium. PMID- 11861516 TI - Antagonists selective for estrogen receptor alpha. AB - To develop compounds that are antagonists on ER(alpha), but not ER(beta), we have added basic side-chains typically found in nonsteroidal antiestrogens to pyrazole compounds that bind with much higher affinity to ER(alpha) than to ER(beta). In this way we have developed basic side-chain pyrazoles (BSC-pyrazoles) that are high affinity, potent, selective antagonists on ER(alpha). These BSC-pyrazoles are themselves inactive on ER(alpha) and ER(beta), and they antagonize E2 stimulation by ER(alpha) only. We investigated seven basic side-chain substituents on various alkyl-triaryl-substituted pyrazoles, and the most ER(alpha)-selective compound was methyl-piperidino-pyrazole (MPP). ER(alpha) selective antagonism was observed on diverse reporter-promoter gene constructs containing estrogen response elements that are consensus, nonconsensus (pS2), or comprised of multiple half-estrogen response elements (NHERF/EBP50) and on genes in which ER works indirectly by tethering to other DNA-bound proteins (TGF(beta)3). In contrast to these BSC-pyrazoles, the antiestrogens trans hydroxytamoxifen, raloxifene, and ICI 182,780 suppress E2 activity via both ER(alpha) and ER(beta). The most effective BSC-pyrazole, MPP, fully antagonized E2 stimulation of pS2 mRNA in MCF-7 breast cancer cells, consistent with the fact that these cells contain almost exclusively ER(alpha). These compounds should be useful in studying the biological functions of ER(alpha) and ER(beta) and in selectively blocking responses that are mediated through ER(alpha). PMID- 11861517 TI - White adipocyte vascular endothelial growth factor: regulation by insulin. AB - White adipose tissue from rats was examined for insulin- responsive vascular endothelial growth factor 165 (VEGF) secretion and mRNA expression. When separated into it constituent fat vs. stromal-vascular cells using collagenase digestion methods, only the adipocytes (or whole fat tissue) responded to physiological insulin concentrations by doubling VEGF release over 4 and 24 h in culture. Adipocyte VEGF mRNA expression increased similarly. Several adipose depots were tested. Although omental fat cells had the highest rates of VEGF release, the differences were not significant. Insulin-stimulated VEGF release was mediated in part via PI3K, but not PKC. Additional hormones/agents were tested, including steroids, leptin, an adenosine analog, and norepinephrine. Only the latter compound increased VEGF production, and this effect was mediated by adenylate cyclase. Adjusting the incubation glucose concentration between 0-20 mM did not alter adipocyte VEGF release. An experimental mimic of hypoxia, CoCl(2), also increased adipocyte VEGF, and this effect was additive with 100 nM insulin. These studies demonstrate that physiological insulin concentrations stimulate VEGF formation and expression in cultured rodent white adipocytes. Although the biological significance of this observation remains to be determined, if white adipocyte-derived VEGF has paracrine or systemic endocrine actions, these might hypothetically impact on adipose expansion or the vascular comorbidities of obesity. PMID- 11861515 TI - Rapid nongenomic E2 effects on p42/p44 MAPK, activator protein-1, and cAMP response element binding protein in rat white adipocytes. AB - In some tissues, rapid effects of estrogens have been described at the plasma membrane level including activation of the MAPK activity. In rat adipocytes, the present study demonstrates that physiological concentrations (0.1-10 nM) of E2 rapidly activate the p42/p44 MAPK. This effect was blocked by the pure estrogen antagonist, ICI 182 780, and appeared specific for E2 because 17alpha-E2, T, and progesterone failed to change the MAPK activity. Pertussis toxin; PP2, a selective inhibitor of Src family kinase; and wortmannin all reduced the magnitude of MAPK activation by E2 suggesting involvement of the Gi-protein/Src family kinase/PI3K pathway. Classical PKCs and MAPK kinase were also involved in MAPK activation by E2. Interestingly, this activation was observed in late but not early differentiated rat preadipocytes, and the immunoreactive ER(alpha) protein was detected only in adipocyte membrane, suggesting that the adipocyte membrane structure is required for the nongenomic effect of E2. Moreover, E2 induced a rapid nuclear translocation of MAPK together with a fast MAPK- dependent activation of cAMP response element binding protein leading to a transcriptional activation of cAMP response element binding protein-responsive genes and reported plasmids. However, the E2 increase in adipocyte activator protein-1 DNA binding does not seem to be fully explained by the E2 activation of the MAPK pathway. This study provides clear evidence for an additional nongenomic mechanism whereby estrogens may exert their control on adipose tissue metabolism. PMID- 11861518 TI - Testosterone (T)-induced changes in arcuate nucleus cocaine-amphetamine-regulated transcript and NPY mRNA are attenuated in old compared to young male brown Norway rats: contribution of T to age-related changes in cocaine-amphetamine-regulated transcript and NPY gene expression. AB - The age-related decrease in serum T levels is associated with impairments in food intake and weight regulation and alterations in brain peptides that regulate energy balance. To test the hypothesis that reduced T levels contribute to altered hypothalamic cocaine-amphetamine-regulated transcript (CART) and NPY gene expression, the mRNA content of these neuropeptides was measured by in situ hybridization in sham-operated (intact), castrated, and T-replaced castrated young and old male Brown Norway rats. T levels in T-replaced young and old rats were similar to those in intact young animals. Compared with castrated rats, arcuate nucleus CART mRNA was lower and NPY mRNA was higher in both young and old T-replaced castrated animals, suggesting reciprocal regulation of these peptides by T; these T-induced changes were localized primarily in the rostral arcuate and were markedly attenuated in old animals. Compared with intact animals, paraventricular nucleus CART mRNA was lower in castrated animals and similar in T replaced young and old rats. We conclude that hypothalamic CART and NPY neurons remain responsive to T regulation in old rats, albeit less so than in young animals, suggesting that the age-related reduction of T contributes in part to altered brain neuropeptide gene expression favoring anorexia and wasting with aging. PMID- 11861520 TI - The goitrogen 6-n-propyl-2-thiouracil (PTU) given during testis development increases Sertoli and germ cell numbers per cyst in fish: the tilapia (Oreochromis niloticus) model. AB - The main objectives of the present study were to investigate the effects of 6-n propyl-2-thiouracil (PTU) on Sertoli cell proliferation, germ cell number, and testis size in Nile tilapias (Oreochromis niloticus). In this regard, young fish (approximately 1 g BW and approximately 3.5 cm total in length) were treated for a period of 40 d with different concentrations (100 and 150 ppm) of PTU. The animals were killed and analyzed on d 1, 30, 40, 98, and 208 after the beginning of the treatment. On d 30 and 40 the spermatogenic process was delayed in fish treated with PTU compared with the control group. Also at these periods, treated tilapia had decreased (P < 0.05) body weight and total length. On d 98 body weight and total length had recovered in PTU-treated fish and were similar (P > 0.05) to those of the controls. However, testis weight and gonadosomatic index (testis mass/body weight) were approximately 100% higher (P < 0.05) in treated tilapia. Similarly, the area occupied by seminiferous tubules, the number of Sertoli cells and germ cells per cyst, and the number of Leydig cells per testis were significantly (P < 0.05) greater in treated fish. Nevertheless, nuclear volume and individual Leydig cell volume were significantly lower (P < 0.05) in tilapia receiving PTU treatment. Compared with controls, at 208 d all parameters analyzed presented the same trend as that observed at 98 d. In general, at 98 d the different PTU concentrations used during the treatment period induced similar effects. However, at 208 d the mean values observed for several parameters were significantly higher (P < 0.05) in fish exposed to 150 ppm. Probably due to the higher density of Sertoli cells per cyst in treated tilapia, these cells presented a smaller (P < 0.05) nucleolus and a trend to decrease its support capacity (efficiency). However, the meiotic index (germ cell loss during the two meiotic divisions) was similar (P > 0.05) in the three groups of fish investigated. Remarkably, the results found in tilapia were similar to those found for rats treated with PTU. This suggests strongly that the mechanisms of control of Sertoli cell and Leydig cell proliferation seem to be preserved during vertebrate evolution. PMID- 11861519 TI - Effect of adenovirus-mediated overexpression of follistatin and extracellular domain of activin receptor type II on gonadotropin secretion in vitro and in vivo. AB - Activins are dimeric proteins that stimulate the synthesis and secretion of pituitary FSH by interacting with two classes of receptors, type I and type II, to initiate their intracellular signaling cascade. The extracellular domain of type II activin receptor (ActRII-ECD) contains all structural determinants sufficient for high affinity ligand binding. A soluble recombinant ActRII-ECD has been reported to attenuate FSH secretion from cultured rat anterior pituitary cells in response to exogenous activin A or endogenous activin B. Follistatin is a binding protein that acts as an extracellular factor to bind and inactivate activin. We constructed adenoviral vectors able to mediate expression of follistatin 288 (AdexCAFS288) and ActRII-ECD (AdexCAECD) and tested their biological activities both in vitro and in vivo. The data show that adenovirus mediated overexpression of either ActRII-ECD or follistatin was able to attenuate FSH secretion by cultured rat anterior pituitary cells. However, AdexCAFS288 overexpression of follistatin was more effective than adenovirus-mediated overexpression of ActRII-ECD. In vivo, a single ip injection of AdexCAFS288 induced the expression of high levels of follistatin and resulted in the suppression of serum FSH levels in castrated male rats for up to 12 d postinjection. Infection with AdexCAFS288 had no effect on LH secretion in vitro or in vivo, demonstrating its selectivity. In conclusion, the results demonstrate the effectiveness of adenovirus-mediated overexpression of follistatin and ActRII ECD to regulate FSH secretion and the potential of using this strategy as a tool to further define the critical role of activin/inhibin/follistatin circuitry in the modulation of the reproductive system. PMID- 11861522 TI - TR(beta)1 protein is preferentially expressed in the pericentral zone of rat liver and exhibits marked diurnal variation. AB - We investigated the distribution and diurnal variation of TR(beta)1 protein expression in liver with specific antibodies against TR(beta)1. Immunohistochemistry showed a zonal distribution of TR(beta)1 with maximum expression in the pericentral zone matching some known T(3)-responsive enzyme activities in the liver, such as glutamine synthetase, cholesterol 7alpha- hydroxylase, and spot 14. Combining immunohistochemistry and image analysis we found and quantified the same zonal distribution for 5'-deiodinase type 1 as for TR(beta)1. Western blot analysis revealed a profound diurnal variation for TR(beta)1 protein expression, with highest levels at the beginning of the dark period. TR(beta)1 diurnal variation partly overlaps with the T(3)-responsive genes, cholesterol 7alpha-hydroxylase and spot 14. Furthermore, TR(beta)1 distribution along the porto-central axis does not change during the day, indicating that the zonal expression of TR(beta)1 is stable. This is the first time that zonal distribution in liver has been demonstrated for a member of the nuclear receptor family. This finding together with the observed diurnal rhythm has major implications for interpreting and timing experiments concerning the TR and its downstream actions in liver. PMID- 11861523 TI - Activin A augments GnRH-mediated transcriptional activation of the mouse GnRH receptor gene. AB - The response of pituitary gonadotropes to GnRH correlates directly with the concentration of GnRH receptors (GnRHRs) on the cell surface, which is mediated in part at the level of GnRHR gene expression. We have previously localized GnRH responsiveness in the mouse GnRHR (mGnRHR) gene promoter to two elements: activating protein-1 and sequence underlying responsiveness to GnRH-1. This study was designed to investigate potential synergy between GnRH and activin A in transcriptional activation of the mGnRHR gene. In functional transfection studies using alphaT3-1 cells, GnRH agonist stimulation of the mGnRHR gene promoter ( 765/+62) resulted in a 10.9-fold increase in activity, which was further increased 2-fold (to 21.3-fold) following activin pretreatment. Activin pretreatment alone had no effect. Deletion of region -387/-308 or mutation of a putative SMAD-binding element at -331/-324 (5'-GTCTAG[T]C-3') abrogated the augmented response to GnRH in the presence of activin but not the response to GnRH alone. Overexpression of SMAD2 and SMAD3 along with SMAD4 increased transcriptional activity of the mGnRHR gene, which was further increased by GnRH agonist stimulation. These data demonstrate that activin augments GnRH-mediated transcriptional activation of the mGnRHR gene and suggest that this effect may be mediated through SMAD transcription factors. PMID- 11861525 TI - Induction of adipocyte complement-related protein of 30 kilodaltons by PPARgamma agonists: a potential mechanism of insulin sensitization. AB - Adipocyte complement-related protein of 30 kDa (Acrp30, adiponectin, or AdipoQ) is a fat-derived secreted protein that circulates in plasma. Adipose tissue expression of Acrp30 is lower in insulin-resistant states and it is implicated in the regulation of in vivo insulin sensitivity. Here we have characterized the ability of PPARgamma agonists to modulate Acrp30 expression. After chronic treatment of obese-diabetic (db/db) mice with PPARgamma agonists (11 d), mean plasma Acrp30 protein levels increased (>3x). Similar effects were noted in a nongenetic type 2 diabetes model (fat-fed and low-dose streptozotocin-treated mice). In contrast, treatment of mice (db/db or fat-fed) with metformin or a PPARalpha agonist did not affect plasma Acrp30 protein levels. In a cohort of normal human subjects, 14-d treatment with rosiglitazone also produced a 130% increase in circulating Acrp30 levels vs. placebo. In addition, circulating Acrp30 levels were suppressed 5-fold in patients with severe insulin resistance in association with dominant-negative PPARgamma mutations. Thus, induction of adipose tissue Acrp30 expression and consequent increases in circulating Acrp30 levels represents a novel potential mechanism for PPARgamma-mediated enhancement of whole-body insulin sensitivity. Furthermore, Acrp30 is likely to be a biomarker of in vivo PPARgamma activation. PMID- 11861526 TI - High glucose levels increase major histocompatibility complex class I gene expression in thyroid cells and amplify interferon-gamma action. AB - Increased major histocompatibility complex (MHC) class I gene expression in target tissues may be relevant to the pathogenesis of autoimmune diseases. In this study, we questioned whether high glucose levels might increase MHC class I levels and thereby contribute to autoimmune complications. We used thyrocytes in continuous culture, because there is an increased incidence of autoimmune thyroiditis in type 2 diabetics and because transcriptional regulation of MHC class I is well studied in these cells. Northern analysis and flow cytometry showed that 20 and 30 mM D-glucose up-regulated MHC class I expression and that the glucose effect was additive to and independent of interferon-gamma. The effect was specific, because L-glucose did not modify class I expression. The glucose acted transcriptionally, requiring both enhancer A and a cAMP-response element-like element located in the hormone-sensitive region of the MHC class I 5'flanking region. These elements are different from those activated by interferon-gamma. High glucose levels increase formation of the MOD-1 complex with enhancer A; MOD-1 is a heterodimer of fra-2 and the p50 subunit of NF kappaB. Both TSH and insulin are required for full expression of the glucose activity in thyrocytes. The glucose effect is partially blocked by wortmannin, suggesting involvement of the PI3K signal system. The data support the possibility that high serum glucose levels in type 2 diabetic patients may increase MHC class I levels in target tissues and contribute to autoimmune complications of the disease. PMID- 11861527 TI - Activation of MAPK cascades by GnRH: ERK and Jun N-terminal kinase are involved in basal and GnRH-stimulated activity of the glycoprotein hormone LHbeta-subunit promoter. AB - The role of ERK and Jun N-terminal kinase (JNK) in basal- and GnRH-stimulated LHbeta-promoter activity was examined in the gonadotroph cell line LbetaT-2. GnRH agonist (GnRH-A) stimulates the MAPK cascades ERK, JNK, and p38MAPK, with a peak at 7 min for ERK and at 60 min for JNK and p38MAPK. The rat glycoprotein hormone LHbeta-subunit promoter, linked to the chloramphenicol acetyl transferase (CAT) reporter gene, was used to follow its activation. Addition of GnRH-A (10 nM) to LbetaT-2 cells resulted in a 6-fold increase in LHbeta-CAT activity at 8 h, which was markedly reduced by a GnRH antagonist. The PKC activator 12-O tetradecanoylphorbol-13-acetate (TPA), but not the Ca(2+) ionophore ionomycin, stimulated LHbeta-CAT activity. Addition of GnRH-A and TPA together did not produce an additive response. Down-regulation of PKC, but not removal of Ca(2+), abolished the GnRH-A and the TPA response. Cotransfection of the LHbeta-promoter and the constitutively active form of Raf-1 stimulated basal and GnRH-A-induced LHbeta-CAT activity. The dominant negative forms of the ERK cascade members Ras, Raf-1, and MAPK/ERK kinase (MEK) markedly reduced basal and GnRH-A-induced LHbeta CAT activity, Similar results were obtained with the MEK inhibitor PD 098059. Cotransfection of the LHbeta-promoter and the constitutively active CDC42 stimulated basal and GnRH-A-induced LHbeta-CAT activity. The dominant negative forms of the JNK cascade members Rac, CDC42, and SEK markedly diminished basal and GnRH-A-induced LHbeta-CAT activity. Interestingly, the constitutively active form of c-Src stimulated the basal and the GnRH-A response, whereas the dominant negative form of c-Src, or the c-Src inhibitor PP1 diminished basal and the GnRH A response. We conclude that ERK and JNK are involved in basal and GnRH-A stimulation of LHbeta-CAT activity. c-Src participates also in LHbeta-promoter activation by a mechanism which might be linked to ERK and JNK activation. PMID- 11861529 TI - MA-10 cells transfected with the human lutropin/choriogonadotropin receptor (hLHR): a novel experimental paradigm to study the functional properties of the hLHR. AB - MA-10 cells are a clonal strain of mouse Leydig tumor cells that retain many of the properties of Leydig cells including expression of the endogenous lutropin/choriogonadotropin receptor (LHR) and the ability to respond to LH/CG with increased steroidogenesis. Recently we noted a dramatic decrease in expression of the endogenous LHR. Although we do not have an explanation for this decline, we took advantage of it to devise a method that allows for the expression of the recombinant human LHR (hLHR) in a Leydig cell model that is now practically devoid of endogenous LHR. We show that the recombinant hLHR can be expressed at variable densities in MA-10 cells and that it can stimulate cAMP and steroid synthesis as well as activate the inositol phosphate and MAPK cascades. We also show that two naturally occurring mutants of the hLHR associated with Leydig cell hyperplasia and one mutant associated with Leydig cell adenomas are constitutively active when assayed for activation of cAMP, inositol phosphate, progesterone, and MAPK. Our ability to express the hLHR in MA-10 cells (now practically devoid of endogenous LHR) provides a novel paradigm to study the cellular and molecular basis of the functions of the LHR in Leydig cells. PMID- 11861530 TI - Targeting of green fluorescent protein to secretory granules in oxytocin magnocellular neurons and its secretion from neurohypophysial nerve terminals in transgenic mice. AB - Oxytocin (OT) is a hypothalamic nonapeptide that is synthesized as part of a larger precursor protein that also contains an approximately 10-kDa protein called neurophysin at its C-terminus. This precursor protein is trafficked through the regulated secretory pathway into secretory granules and then axonally transported to and secreted from nerve terminals in the neural lobe of the pituitary. In this paper, we show that the AI-03 transgene that contains enhanced green fluorescent protein (EGFP) fused to the end of the neurophysin at the C terminus of the OT pre-prohormone, is expressed selectively in OT-magnocellular neurons and is trafficked to secretory granules in transgenic mice. The EGFP containing secretory granules are then transported to OT-neurosecretory terminals in the neurohypophysis, where the EGFP fluorescence undergoes depolarization induced calcium-dependent secretion. The endogenous fluorescence in the neural lobes is sufficiently intense to image secretory events in individual OT nerve terminals (neurosecretosomes) isolated from the posterior pituitaries in these transgenic mice. PMID- 11861531 TI - Identification and characterization of the murine and human gene encoding the tuberoinfundibular peptide of 39 residues. AB - By screening public databases, we identified human and mouse genomic DNA clones that encode the tuberoinfundibular peptide of 39 residues (TIP39). The TIP39 precursor is encoded by at least three exons; a noncoding exon U1, exon 1 encoding residues -61 (initiator methionine) to -19 of the leader sequence, and exon 2 encoding residues -18 to -1 and residues +1 to +39. Secreted human TIP39 is identical to the previously isolated bovine TIP39, whereas mouse TIP39 differs by four amino acids. Phylogenetic analyses suggested that TIP39, PTH, and PTHrP may have evolved from a common ancestor. Synthetic human and mouse TIP39 showed indistinguishable potencies [EC(50): 0.54 (human) vs. 0.74 nM (mouse)] at the human PTH2-receptor stably expressed in LLCPK(1) cells; furthermore, TIP-(9-39) was an inhibitor of cAMP accumulation stimulated by either [Tyr(34)]PTH(1 34)amide or human/bovine TIP39. In the mouse, an approximately 4.5-kb mRNA encoding TIP39 was identified by Northern blot analysis in testis and, less abundantly, in liver and kidney, whereas other tissues revealed additional smaller transcripts. In situ hybridizations revealed TIP39 expression in seminiferous tubuli and several brain regions, including nucleus ruber, nucleus centralis pontis, and nucleus subparafascicularis thalami. Because PTH2 receptor expression was previously shown to be highest in brain, pancreas, and testis, our findings are consistent with the notion that TIP39 is a neuropeptide which may also have a role in spermatogenesis. PMID- 11861533 TI - 17beta-estradiol affords protection against 4-vinylcyclohexene diepoxide-induced ovarian follicle loss in Fischer-344 rats. AB - Repeated dosing with 4-vinylcyclohexene diepoxide (VCD) accelerates atresia via apoptosis in primordial and primary follicles in ovaries of rats. The mechanisms that control atresia and VCD-induced toxicity are unknown; however, they could involve 17beta-E2. Atresia slows as animals enter puberty, whereas circulating E2 levels increase with the the onset of cyclicity. This inverse relationship suggests that E2 may be involved in the control of atresia. Therefore, this study was designed to determine whether treatment of immature rats with E2 could protect follicles normally destroyed by VCD-induced apoptosis. Female F344 rats were treated daily with E2, ER analogs, and/or VCD for 15 d. VCD alone caused a 50% reduction in primordial and primary follicles. Coinjection of E2 (0.1 mg/kg) and VCD (80 mg/kg) selectively protected primary follicles from VCD-induced follicle loss. This protection was mimicked by an ER agonist, genistein (0.1 mg/kg), and prevented by an ER antagonist, 4-hydroxytamoxifen (2 mg/kg). VCD treatment increased caspase-3-like activity, whereas concurrent treatment with genistein and VCD restored caspase-3-like activity to control levels. VCD treatment had no effect on circulating E2 levels, uterine weight, or E2 binding to the ER, nor could it directly displace E2 from ERbeta. These observations support the idea that ER-mediated protection against VCD-induced follicle toxicity is obtained by reducing apoptosis in small preantral follicles, although VCD does not appear to directly interact with ER. PMID- 11861534 TI - The distribution of betaglycan protein and mRNA in rat brain, pituitary, and gonads: implications for a role for betaglycan in inhibin-mediated reproductive functions. AB - Betaglycan was reported by our laboratory to serve as an inhibin binding protein and to facilitate the antagonism of activin signaling. Although an accessory receptor for TGFbeta and inhibin, its distribution within reproductive tissues remains largely unexplored. Histochemical analyses reveal betaglycan protein and mRNA distributed throughout the rat reproductive axis. In the brain, betaglycan mRNA is localized in discrete regions of the forebrain and brain stem, including olfactory, septal, and hypothalamic nuclei. In the pituitary, moderate levels of betaglycan protein and mRNA were observed in the anterior and intermediate lobes. Betaglycan immunoreactivity was colocalized with all the pituitary cell subtypes, to the greatest extent with the gonadotrope population. In the gonads, betaglycan mRNA was localized in cellular compartments, coinciding with its protein for the most part. Moderate levels of mRNA were observed in ovarian granulosa cells, with lower expression in the thecal layer and the oocyte. In the testes, betaglycan mRNA was observed in the Leydig and tubule-specific germ cells. This is the first comprehensive report detailing the distribution of betaglycan in mammalian reproductive tissues. The present findings illustrate and support the hypothesis of a modulatory role for betaglycan in TGFbeta and/or inhibin effects in these tissues. PMID- 11861535 TI - Anti-Mullerian hormone inhibits initiation of primordial follicle growth in the mouse ovary. AB - Recruitment of primordial follicles is essential for female fertility; however, the exact mechanisms regulating this process are largely unknown. Earlier studies using anti-Mullerian hormone (AMH)-deficient mice suggested that AMH is involved in the regulation of primordial follicle recruitment. We tested this hypothesis in a neonatal ovary culture system, in which ovaries from 2-d-old C57Bl/6J mice were cultured for 2 or 4 d in the absence or presence of AMH. Ovaries from 2-d old mice contain multiple primordial follicles, some naked oocytes, and no follicles at later stages of development. We observed that in the cultured ovaries, either nontreated or AMH-treated, follicular development progressed to the same extent as in in vivo ovaries of comparable age, confirming the validity of our culture system. However, in the presence of AMH, cultured ovaries contained 40% fewer growing follicles compared with control ovaries. A similar reduction was found after 4 d of culture. Consistent with these findings, we noted lower inhibin alpha-subunit expression in AMH-treated ovaries compared with untreated ovaries. In contrast, expression of AMH ligand type II receptor and the expression of oocyte markers growth and differentiation factor 9 and zona pellucida protein 3 were not influenced by AMH. Based on the results, we suggest that AMH inhibits initiation of primordial follicle growth and therefore functions as an inhibitory growth factor in the ovary during these early stages of folliculogenesis. PMID- 11861536 TI - Repression of the steroidogenic acute regulatory gene by the multifunctional transcription factor Yin Yang 1. AB - The transport of cholesterol to the inner mitochondrial membrane by the steroidogenic acute regulatory (StAR) protein is a critical step in steroidogenesis. The current study was designed to examine whether the multifunctional transcription factor Yin Yang 1 (YY1) was able to repress activation of the StAR gene. YY1 bound to three putative YY1-binding sites in the rat StAR promoter. Cotransfection of the StAR promoter linked to a luciferase reporter gene and YY1 in the presence or absence of sterol regulatory element binding protein-1a (SREBP-1a) resulted in transcriptional repression. YY1 was found to colocalize in the nucleus with SREPB-1a. YY1 inhibited SREBP-1a/nuclear factor Y (NF-Y) enhancement of StAR activation and YY1 decreased SREBP-1a binding to a sterol regulatory element in the presence or absence of NF-Y. YY1 also decreased NF-Y binding to a nonconsensus NF-Y-binding motif in the StAR promoter. These studies provide novel information on the mechanisms of YY1-mediated repression of the StAR gene. PMID- 11861538 TI - TNFalpha potently activates osteoclasts, through a direct action independent of and strongly synergistic with RANKL. AB - TNFalpha is pivotal to the pathogenesis of inflammatory and possibly postmenopausal osteolysis. Much recent work has clarified mechanisms by which TNFalpha promotes osteoclastogenesis, but the means by which it activates osteoclasts to resorb bone remain uncertain. We found that very low concentrations of TNFalpha promoted actin ring formation, which correlates with functional activation in osteoclasts, both in osteoclasts formed in vitro and extracted from newborn rats. TNFalpha was equipotent with RANKL for this action. Activation by TNFalpha was unaffected by blockade of RANKL by OPG, its soluble decoy receptor, suggesting that this was due to a direct action on osteoclasts. Bone resorption was similarly directly and potently stimulated, in a RANKL independent manner in osteoclasts, whether these were formed in vitro or in vivo. Interestingly, TNFalpha promoted actin ring formation at concentrations an order of magnitude below those required for osteoclastic differentiation. Moreover, TNFalpha strongly synergized with RANKL, such that miniscule concentrations of TNFalpha were sufficient to substantially augment osteoclast activation. The extreme sensitivity of osteoclasts to activation by TNFalpha suggests that the most sensitive osteolytic response of bone to TNFalpha is through activation of existing osteoclasts; and the strong synergy with RANKL provides a mechanism whereby increased osteolysis can be achieved without disturbance to the underlying pattern of osteoclastic localization. PMID- 11861537 TI - Distribution, neuronal colocalization, and 17beta-E2 modulation of small conductance calcium-activated K(+) channel (SK3) mRNA in the guinea pig brain. AB - Molecular cloning has revealed the existence of three distinct small conductance (SK1-3) Ca(2+)-activated K(+) channels. Because SK channels underlie the afterhyperpolarization (AHP) that is critical for sculpturing phasic firing in hypothalamic neurons, we investigated the distribution of these channels in the female guinea pig. Both SK1 and SK3 cDNA fragments were cloned using PCR, and ribonuclease protection assay as well as in situ hybridization analysis illustrated that the SK3 channel was the predominant subtype expressed in the guinea pig hypothalamus. Combined in situ hybridization and fluorescence immunocytochemistry revealed that SK3 mRNA was expressed in GnRH, dopamine, and vasopressin neurons, and all of these neurons exhibited an AHP current. Moreover, SK3 mRNA was found in other brain areas, including the septum, bed nucleus, amygdala, thalamus, midbrain, and hippocampus. Using quantitative ribonuclease protection assay, the rank order of SK3 mRNA expression was septum >or= midbrain > rostral thalamus >or= rostral basal hypothalamus >or= caudal thalamus >or= preoptic area >> caudal basal hypothalamus >or= hippocampus. Moreover, 17beta-E2 treatment, which reduces plasma LH during the negative feedback phase, significantly increased SK3 mRNA levels in the rostral basal hypothalamus (P < 0.05; n = 6). Therefore, these findings suggest that estrogen increases the mRNA expression of SK3 channels, which may represent a mechanism by which estrogen regulates hypothalamic neuronal excitability during negative feedback. PMID- 11861539 TI - Testosterone (T) enhances apoptosis-related damage in human vascular endothelial cells. AB - Androgens may contribute to higher cardiovascular risk in men via deleterious effects on vascular endothelial cells (EC). We examined the effects of androgens on male human umbilical vein EC (EA.hy926) in culture. [(3)H]Thymidine incorporation assays showed that after 24-h serum deprivation, testosterone (T) (but not dehydroepiandrosterone nor 17beta-E2) induced significant dose-dependent decreases in DNA synthesis (10-16% at 1-100 nmol/liter); the AR antagonist flutamide (100 nmol/liter) abolished this effect of T. After 48-h serum deprivation, typical apoptotic DNA patterns were detected in agarose gels, and the number of floating cells indicative of severe damage was significantly greater after T treatment for 48 and 72 h (13.7 +/- 0.5% and 30.2 +/- 2.5%, respectively) than the control values (9.7 +/- 1.05% and 23.7 +/- 3.0%). Analysis of attached cells by annexin V-fluorescein isothiocyanate/propidium iodide staining showed that after 48-h serum deprivation, T significantly increased the number of cells in the early (16.0 +/- 1.1%) and late (8.3 +/- 0.3%) stages of apoptosis compared with control (6.8 +/- 1.0% and 4.0 +/- 0.2%, respectively); such increases in apoptosis-related damage were also observed, to a lesser degree, in serum-enriched culture. Western blotting showed that B cell leukemia/lymphoma-2 protein (Bcl-2) expression decreased significantly in serum deprived EC treated with T. Thus, T reduces DNA synthesis and enhances apoptosis after serum deprivation in EC, possibly related to reduced Bcl-2 expression. PMID- 11861540 TI - PRL gene expression in individual living mammotropes displays distinct functional pulses that oscillate in a noncircadian temporal pattern. AB - PRL gene expression in the anterior pituitary has been the focus of intensive investigation for many years, but very little information is available on the actual dynamics by which this process occurs in individual mammotrope cells. Here, we used single cell bioluminescent imaging microscopy and a recently refined reporter gene strategy to measure PRL promoter-driven gene expression (PRL-GE) in individual living primary mammotropes. Using this approach we report a new phenomenon involving repetitive on/off gene expression bursts that occurred in a distinctly noncircadian oscillatory pattern. Furthermore, we demonstrate a functional basis for these gene expression oscillations, inasmuch as PRL-GE pulses were sensitive to calcium-dependent modulation, which we show arose exclusively as changes in the shape of individual pulse episodes. Our results provide the first clear evidence that PRL-GE, in its homologous cell environment, displays oscillatory bursts of activity. Moreover, they strongly support the idea that these discrete on/off bursts of activity serve as an important determinant of the timing and level of PRL-GE under both basal and stimulated conditions. PMID- 11861541 TI - Molecular characterization of canine prostaglandin G/H synthase-2 and regulation in prostatic adenocarcinoma cells in vitro. AB - Induction of PG G/H synthase-2 (PGHS-2), a key rate-limiting enzyme in the PG biosynthetic pathway, has been implicated in prostatic adenocarcinomas in humans and dogs in vivo, but the molecular control of PGHS-2 expression in prostate cancer remains poorly understood. Using the dog model, the specific objectives of this study were to clone and characterize canine PGHS-2, and to study the regulation of its transcript, protein, and activity in a canine prostatic adenocarcinoma (CPA) cell line in vitro. The canine PGHS-2 cDNA was cloned by a combination of cDNA library screening and 5'-rapid amplification of cDNA ends, and shown to contain a 5'-untranslated region of 28 bp, an open reading frame of 1815 bp, and a 3'-untranslated region of 1655 bp. The open reading frame encodes a 604-amino acid protein that is 89% identical to the human homolog. The regulation of PGHS-2 protein and PGE(2) synthesis was studied in CPA cells cultured in the absence or presence of graded doses of phorbol 12-myristate 13 acetate (PMA), TNFalpha, and lipopolysaccharides. Results from immunoblots, immunocytochemistry, and RIAs showed that PGHS-2 protein and PGE(2) were present at low levels in control cells and were significantly induced after agonist treatment (P < 0.05), with PMA being the strongest inducer. Northern blot analyses also revealed a significant increase of PGHS-2 mRNA by PMA, TNFalpha, and lipopolysaccharides treatment (P < 0.05). Agonist-dependent induction of PGHS 2 mRNA was not dependent on new protein synthesis (coincubation with cycloheximide; 10 microg/ml) but was blocked by transcription inhibitor actinomycin D (5 microg/ml), suggesting that PGHS-2 acts an immediate early response gene in prostatic epithelial cells. Thus, this study characterizes for the first time the structure of canine PGHS-2 and provides an in vitro model to unravel the molecular basis of PGHS-2 expression in prostatic adenocarcinomas. PMID- 11861542 TI - Structure and dynamics of the fusion pores in live GH-secreting cells revealed using atomic force microscopy. AB - Earlier studies in live pancreatic acinar cells identified new cellular structures at the cell plasma membrane called 'pits' and 'depressions', where membrane-bound secretory vesicles dock and fuse to release vesicular contents. In the current study, using atomic force microscopy we identify similar structures at the plasma membrane of GH-secreting cells of the pituitary and implicate their involvement in hormone release. Pits containing 100-200 nm in diameter depressions or fusion pores were identified in resting GH-secreting cells. Following stimulation of secretion the size of depression enlarged and gold tagged GH antibody were found to bind to the pit structures in the stimulated GH cells. This study documents for the first time the presence of these structures and their involvement in secretions in a neuroendocrine cell. PMID- 11861545 TI - The emergence of hymenopteran genetics. PMID- 11861546 TI - Structure/function analysis of the Saccharomyces cerevisiae Trf4/Pol sigma DNA polymerase. AB - The Trf4p/Pol sigma DNA polymerase (formerly Trf4p/Pol kappa) couples DNA replication to the establishment of sister chromatid cohesion. The polymerase is encoded by two redundant homologs in Saccharomyces cerevisiae, TRF4 and TRF5, that together define a fourth essential nuclear DNA polymerase in yeast and probably in all eukaryotes. Here we present a thorough genetic analysis of the founding member of this novel family of DNA polymerases, TRF4. Analyses of mutants carrying 1 of 34 "surface-targeted" alanine scanning mutations in TRF4 have identified those regions required for Pol sigma's essential function, for its role in DNA double-strand break repair, and for its association with chromosomes. The data strongly support the importance of the regions of predicted structural similarity with the Pol beta superfamily as critical for Trf4p/Pol sigma's essential and repair functions. Surprisingly, five lethal mutations lie outside all polymerase homology in a C-terminal region. The protein possesses Mg2+-dependent 3' to 5' exonuclease activity. Cell cycle analysis reveals that Trf4p/Pol sigma associates with chromosomes in G1, S, and G2 phases, but that association is abolished coincident with dissolution of cohesion at the metaphase to-anaphase transition. PMID- 11861547 TI - Genetic interactions of yeast eukaryotic translation initiation factor 5A (eIF5A) reveal connections to poly(A)-binding protein and protein kinase C signaling. AB - The highly conserved eukaryotic translation initiation factor eIF5A has been proposed to have various roles in the cell, from translation to mRNA decay to nuclear protein export. To further our understanding of this essential protein, three temperature-sensitive alleles of the yeast TIF51A gene have been characterized. Two mutant eIF5A proteins contain mutations in a proline residue at the junction between the two eIF5A domains and the third, strongest allele encodes a protein with a single mutation in each domain, both of which are required for the growth defect. The stronger tif51A alleles cause defects in degradation of short-lived mRNAs, supporting a role for this protein in mRNA decay. A multicopy suppressor screen revealed six genes, the overexpression of which allows growth of a tif51A-1 strain at high temperature; these genes include PAB1, PKC1, and PKC1 regulators WSC1, WSC2, and WSC3. Further results suggest that eIF5A may also be involved in ribosomal synthesis and the WSC/PKC1 signaling pathway for cell wall integrity or related processes. PMID- 11861548 TI - ATP-dependent remodeling of the spliceosome: intragenic suppressors of release defective mutants of Saccharomyces cerevisiae Prp22. AB - The essential splicing factor Prp22 is a DEAH-box helicase that catalyzes the release of mRNA from the spliceosome. ATP hydrolysis by Prp22 is necessary but not sufficient for spliceosome disassembly. Previous work showed that mutations in motif III (635SAT637) of Prp22 that uncouple ATP hydrolysis from spliceosome disassembly lead to severe cold-sensitive (cs) growth defects and to impaired RNA unwinding activity in vitro. The cs phenotype of S635A (635AAT) can be suppressed by intragenic mutations that restore RNA unwinding. We now report the isolation and characterization of new intragenic mutations that suppress the cold-sensitive growth phenotypes of the T637A motif III mutation (SAA), the H606A mutation in the DEAH-box (DEAA), and the R805A mutation in motif VI (804QAKGRAGR811). Whereas the T637A and H606A proteins are deficient in releasing mRNA from the spliceosome at nonpermissive temperature in vitro, the suppressor proteins have recovered mRNA release activity. To address the mechanisms of suppression, we tested ATPase and helicase activities of Prp22 suppressor mutant proteins and found that the ability to unwind a 25-bp RNA duplex was not restored in every case. This finding suggests that release of mRNA from the spliceosome is less demanding than unwinding of a 25-bp duplex RNA; the latter reaction presumably reflects the result of several successive cycles of ATP binding, hydrolysis, and unwinding. Increasing the reaction temperature allows H606A and T637A to effect mRNA release in vitro, but does not restore RNA unwinding by T637A. PMID- 11861549 TI - Genetic control of extracellular protease synthesis in the yeast Yarrowia lipolytica. AB - Depending on the pH of the growth medium, the yeast Yarrowia lipolytica secretes an acidic protease or an alkaline protease, the synthesis of which is also controlled by carbon, nitrogen, and sulfur availability, as well as by the presence of extracellular proteins. Previous results have indicated that the alkaline protease response to pH was dependent on YlRim101p, YlRim8p/YlPalF, and YlRim21p/YlPalH, three components of a conserved pH signaling pathway initially described in Aspergillus nidulans. To identify other partners of this response pathway, as well as pH-independent regulators of proteases, we searched for mutants that affect the expression of either or both acidic and alkaline proteases, using a YlmTn1-transposed genomic library. Four mutations affected only alkaline protease expression and identified the homolog of Saccharomyces cerevisiae SIN3. Eighty-nine mutations affected the expression of both proteases and identified 10 genes. Five of them define a conserved Rim pathway, which acts, as in other ascomycetes, by activating alkaline genes and repressing acidic genes at alkaline pH. Our results further suggest that in Y. lipolytica this pathway is active at acidic pH and is required for the expression of the acidic AXP1 gene. The five other genes are homologous to S. cerevisiae OPT1, SSY5, VPS28, NUP85, and MED4. YlOPT1 and YlSSY5 are not involved in pH sensing but define at least a second protease regulatory pathway. PMID- 11861550 TI - The cytoplasmic end of transmembrane domain 3 regulates the activity of the Saccharomyces cerevisiae G-protein-coupled alpha-factor receptor. AB - The binding of alpha-factor to its receptor (Ste2p) activates a G-protein signaling pathway leading to conjugation of MATa cells of the budding yeast S. cerevisiae. We conducted a genetic screen to identify constitutively activating mutations in the N-terminal region of the alpha-factor receptor that includes transmembrane domains 1-5. This approach identified 12 unique constitutively activating mutations, the strongest of which affected polar residues at the cytoplasmic ends of transmembrane domains 2 and 3 (Asn84 and Gln149, respectively) that are conserved in the alpha-factor receptors of divergent yeast species. Targeted mutagenesis, in combination with molecular modeling studies, suggested that Gln149 is oriented toward the core of the transmembrane helix bundle where it may be involved in mediating an interaction with Asn84. These residues appear to play specific roles in maintaining the inactive conformation of the protein since a variety of mutations at either position cause constitutive receptor signaling. Interestingly, the activity of many mammalian G-protein coupled receptors is also regulated by conserved polar residues (the E/DRY motif) at the cytoplasmic end of transmembrane domain 3. Altogether, the results of this study suggest a conserved role for the cytoplasmic end of transmembrane domain 3 in regulating the activity of divergent G-protein-coupled receptors. PMID- 11861552 TI - Dimorphic mating-type chromosomes in the fungus Microbotryum violaceum. AB - Fungi often mate as haploids, and sex chromosomes (i.e., mating-type chromosomes) that are dimorphic for their size or overall DNA content have never been reported in this kingdom. Using electrophoretic techniques for karyotype analysis, a highly dimorphic chromosome pair that determines mating compatibility is shown to occur in populations of the fungus Microbotryum violaceum. This substantiates the evolution of such dimorphism as a general feature associated with haploid determination of mating compatibility, which previously had been known only in haplodioecious plants (mosses and liverworts). Size-dimorphic sex chromosomes are present in a lineage of M. violaceum native to Europe, as well as a lineage native to North America. However, they are very different in size between these lineages, indicating either independent evolution of the dimorphism or a large degree of divergence since their isolation. Several DNA sequences that show sequence similarity to transposons were isolated from these sex chromosomes. PMID- 11861551 TI - Schizosaccharomyces pombe Bir1p, a nuclear protein that localizes to kinetochores and the spindle midzone, is essential for chromosome condensation and spindle elongation during mitosis. AB - The inhibitor of apoptosis (IAP) family of proteins contains a subset of members characterized by the presence of highly conserved baculoviral IAP repeat (BIR) domains. Recent work has shown that some of these BIR-domain proteins play a prominent role in the regulation of cell division, in particular at the stage of chromosome segregation and cytokinesis. We and others have shown that the Schizosaccharomyces pombe BIR-domain protein, Bir1p/Pbh1p/Cut17p, is important for the regulation of mitosis. Here we further characterize S. pombe Bir1p using methods of cell biology and genetics. We show that Bir1p is dispersed throughout the nucleus during the cell cycle. In addition, a significant part of Bir1p is also detected at the kinetochores and the spindle midzone during mitosis and meiosis. Time-lapse microscopy studies suggest that Bir1p relocates from the kinetochores to the spindle at the end of anaphase A. Bir1p colocalizes with the S. pombe Aurora kinase homolog Aim1p, a protein essential for mitosis, at the kinetochores as well as the spindle midzone during mitosis, and functional Bir1p is essential for localization of Aim1p to the kinetochores and the spindle midzone. Analyses of bir1 conditional mutants revealed that Bir1p is essential for chromosome condensation during mitosis. In addition, anaphase cells show the presence of lagging chromosomes and a defect in spindle elongation. We conclude that Bir1p is important for multiple processes that occur during mitosis in S. pombe. PMID- 11861553 TI - RNA interference in the pathogenic fungus Cryptococcus neoformans. AB - Cryptococcus neoformans is a pathogenic fungus responsible for serious disease in immunocompromised individuals. This organism has recently been developed as an experimental system, with initiation of a genome project among other molecular advances. However, investigations of Cryptococcus are hampered by the technical difficulty of specific gene replacements. RNA interference, a process in which the presence of double-stranded RNA homologous to a gene of interest results in specific degradation of the corresponding message, may help solve this problem. We have shown that expression of double-stranded RNA corresponding to portions of the cryptococcal CAP59 and ADE2 genes results in reduced mRNA levels for those genes, with phenotypic consequences similar to that of gene disruption. The two genes could also be subjected to simultaneous interference through expression of chimeric double-stranded RNA. Specific modulation of protein expression through introduction of double-stranded RNA thus operates in C. neoformans, which is the first demonstration of this technique in a fungal organism. Use of RNA interference in Cryptococcus should allow manipulation of mRNA levels for functional analysis of genes of interest and enable efficient exploration of genes discovered by genome sequencing. PMID- 11861554 TI - Roles for Caenorhabditis elegans rad-51 in meiosis and in resistance to ionizing radiation during development. AB - We have investigated the role of Caenorhabditis elegans RAD-51 during meiotic prophase and embryogenesis, making use of the silencing effect of RNA interference (RNAi). rad-51 RNAi leads to severe defects in chromosome morphology in diakinesis oocytes. We have explored the effect of rad-51 RNAi in mutants lacking fundamental components of the recombination machinery. If double-strand breaks are prevented by spo-11 mutation, rad-51 RNAi does not affect chromosome appearance. This is consistent with a role for RAD-51 downstream of the initiation of recombination. In the absence of MRE-11, as in the absence of SPO 11, RAD-51 depletion has no effect on the chromosomes, which appear intact, thus indicating a role for MRE-11 in DSB induction. Intriguingly, rad-51 silencing in oocytes that lack MSH-5 leads to chromosome fragmentation, a novel trait that is distinct from that seen in msh-5 mutants and in rad-51 RNAi oocytes, suggesting new potential roles for the msh-5 gene. Silencing of the rad-51 gene also causes a reduction in fecundity, which is suppressed by mutation in the DNA damage checkpoint gene rad-5, but not in the cell death effector gene ced-3. Finally, RAD-51 depletion is also seen to affect the soma, resulting in hypersensitivity to ionizing radiation in late embryogenesis. PMID- 11861555 TI - Caenorhabditis elegans lin-45 raf is essential for larval viability, fertility and the induction of vulval cell fates. AB - The protein kinase Raf is an important signaling protein. Raf activation is initiated by an interaction with GTP-bound Ras, and Raf functions in signal transmission by phosphorylating and activating a mitogen-activated protein (MAP) kinase kinase named MEK. We identified 13 mutations in the Caenorhabditis elegans lin-45 raf gene by screening for hermaphrodites with abnormal vulval formation or germline function. Weak, intermediate, and strong loss-of-function or null mutations were isolated. The phenotype caused by the most severe mutations demonstrates that lin-45 is essential for larval viability, fertility, and the induction of vulval cell fates. The lin-45(null) phenotype is similar to the mek 2(null) and mpk-1(null) phenotypes, indicating that LIN-45, MEK-2, and MPK-1 ERK MAP kinase function in a predominantly linear signaling pathway. The lin-45 alleles include three missense mutations that affect the Ras-binding domain, three missense mutations that affect the protein kinase domain, two missense mutations that affect the C-terminal 14-3-3 binding domain, three nonsense mutations, and one small deletion. The analysis of the missense mutations indicates that Ras binding, 14-3-3-binding, and protein kinase activity are necessary for full Raf function and suggests that a 14-3-3 protein positively regulates Raf-mediated signaling during C. elegans development. PMID- 11861556 TI - Patterns of genetic variation at a chromosome 4 locus of Drosophila melanogaster and D. simulans. AB - DNA sequence surveys of Drosophila melanogaster populations show a strong positive correlation between the recombination rate experienced by a locus and its level of nucleotide polymorphism. In particular, surveys of the fourth chromosome gene ci(D) show greatly reduced levels of nucleotide variation; this observation was originally interpreted in terms of selective sweeps occurring on the nonrecombining fourth chromosome. Subsequent theoretical work has, however, uncovered several other selective processes that can reduce variation. In this study, we revisit the Drosophila fourth chromosome, investigating variation in 5 6 kb of the gene ankyrin in D. melanogaster and D. simulans. Silent nucleotide site diversity is approximately 5 x 10(-4) for both species, consistent with the previous observations of low variation at ci(D). Given the observed frequency spectra at ankyrin, coalescent simulations indicate that reduced diversity in the region is unlikely to be due to a selective sweep alone. We find evidence for recombinational exchange at this locus, and both species appear to be fixed for an insertion of the transposable element HB in an intron of ankyrin. PMID- 11861558 TI - Efficient mobilization of mariner in vivo requires multiple internal sequences. AB - Aberrant products of mariner excision that have an impaired ability to be mobilized often include internal deletions that do not encroach on either of the inverted repeats. Analysis of 13 such deletions, as well as 7 additional internal deletions obtained by various methods, has revealed at least three internal regions whose integrity is necessary for efficient mariner mobilization. Within the 1286-bp element, the essential regions are contained in the intervals bounded by coordinates 229-586, 735-765, and 939-1066, numbering in base pairs from the extreme 5' end of the element. These regions may contain sequences that are necessary for transposase binding or that are needed to maintain proper spacing between binding sites. The isolation of excision-defective elements with point mutations at nucleotide positions 993 and 161/179 supports the hypothesis of sequence requirements, but the reduced mobility of transformation vectors with insertions into the SacI site at position 790 supports the hypothesis of spacing requirements. The finding of multiple internal regions that are essential for efficient mariner mobilization in vivo contrasts with reports that mini-elements with as little as 43 bp of DNA between the inverted repeats can transpose efficiently in vitro. PMID- 11861557 TI - Cytogenetic analysis of the third chromosome heterochromatin of Drosophila melanogaster. AB - Previous cytological analysis of heterochromatic rearrangements has yielded significant insight into the location and genetic organization of genes mapping to the heterochromatin of chromosomes X, Y, and 2 of Drosophila melanogaster. These studies have greatly facilitated our understanding of the genetic organization of heterochromatic genes. In contrast, the 12 essential genes known to exist within the mitotic heterochromatin of chromosome 3 have remained only imprecisely mapped. As a further step toward establishing a complete map of the heterochomatic genetic functions in Drosophila, we have characterized several rearrangements of chromosome 3 by using banding techniques at the level of mitotic chromosome. Most of the rearrangement breakpoints were located in the dull fluorescent regions h49, h51, and h58, suggesting that these regions correspond to heterochromatic hotspots for rearrangements. We were able to construct a detailed cytogenetic map of chromosome 3 heterochromatin that includes all of the known vital genes. At least 7 genes of the left arm (from l(3)80Fd to l(3)80Fj) map to segment h49-h51, while the most distal genes (from l(3)80Fa to l(3)80Fc) lie within the h47-h49 portion. The two right arm essential genes, l(3)81Fa and l(3)81Fb, are both located within the distal h58 segment. Intriguingly, a major part of chromosome 3 heterochromatin was found to be "empty," in that it did not contain either known genes or known satellite DNAs. PMID- 11861560 TI - Sex determination signals control ovo-B transcription in Drosophila melanogaster germ cells. AB - Nonautonomous inductive signals from the soma and autonomous signals due to a 2X karyotype determine the sex of Drosophila melanogaster germ cells. These two signals have partially overlapping influences on downstream sex determination genes. The upstream OVO-B transcription factor is required for the viability of 2X germ cells, regardless of sexual identity, and for female germline sexual identity. The influence of inductive and autonomous signals on ovo expression has been controversial. We show that ovo-B is strongly expressed in the 2X germ cells in either a male or a female soma. This indicates that a 2X karyotype controls ovo-B expression in the absence of inductive signals from the female soma. However, we also show that female inductive signals positively regulate ovo-B transcription in the 1X germ cells that do not require ovo-B function. Genetic analysis clearly indicates that inductive signals from the soma are not required for ovo-B function in 2X germ cells. Thus, while somatic inductive signals and chromosome karyotype have overlapping regulatory influences, a 2X karyotype is a critical germline autonomous determinant of ovo-B function in the germline. PMID- 11861559 TI - Unexpected stability of mariner transgenes in Drosophila. AB - A number of mariner transformation vectors based on the mauritiana subfamily of transposable elements were introduced into the genome of Drosophila melanogaster and examined for their ability to be mobilized by the mariner transposase. Simple insertion vectors were constructed from single mariner elements into which exogenous DNA ranging in size from 1.3 to 4.5 kb had been inserted; composite vectors were constructed with partial or complete duplications of mariner flanking the exogenous DNA. All of the simple insertion vectors showed levels of somatic and germline excision that were at least 100-fold lower than the baseline level of uninterrupted mariner elements. Although composite vectors with inverted duplications were unable to be mobilized at detectable frequencies, vectors with large direct duplications of mariner could be mobilized. A vector consisting of two virtually complete elements flanking exogenous DNA yielded a frequency of somatic eye-color mosaicism of approximately 10% and a frequency of germline excision of 0.04%. These values are far smaller than those observed for uninterrupted elements. The results imply that efficient mobilization of mariner in vivo requires the presence and proper spacing of sequences internal to the element as well as the inverted repeats. PMID- 11861561 TI - The Drosophila gene taranis encodes a novel trithorax group member potentially linked to the cell cycle regulatory apparatus. AB - Genes of the Drosophila Polycomb and trithorax groups (PcG and trxG, respectively) influence gene expression by modulating chromatin structure. Segmental expression of homeotic loci (HOM) initiated in early embryogenesis is maintained by a balance of antagonistic PcG (repressor) and trxG (activator) activities. Here we identify a novel trxG family member, taranis (tara), on the basis of the following criteria: (i) tara loss-of-function mutations act as genetic antagonists of the PcG genes Polycomb and polyhomeotic and (ii) they enhance the phenotypic effects of mutations in the trxG genes trithorax (trx), brahma (brm), and osa. In addition, reduced tara activity can mimic homeotic loss of-function phenotypes, as is often the case for trxG genes. tara encodes two closely related 96-kD protein isoforms (TARA-alpha/-beta) derived from broadly expressed alternative promoters. Genetic and phenotypic rescue experiments indicate that the TARA-alpha/-beta proteins are functionally redundant. The TARA proteins share evolutionarily conserved motifs with several recently characterized mammalian nuclear proteins, including the cyclin-dependent kinase regulator TRIP-Br1/p34(SEI-1), the related protein TRIP-Br2/Y127, and RBT1, a partner of replication protein A. These data raise the possibility that TARA alpha/-beta play a role in integrating chromatin structure with cell cycle regulation. PMID- 11861562 TI - The Drosophila inebriated-encoded neurotransmitter/osmolyte transporter: dual roles in the control of neuronal excitability and the osmotic stress response. AB - Water reabsorption by organs such as the mammalian kidney and insect Malpighian tubule/hindgut requires a region of hypertonicity within the organ. To balance the high extracellular osmolarity, cells within these regions accumulate small organic molecules called osmolytes. These osmolytes can accumulate to a high level without toxic effects on cellular processes. Here we provide evidence consistent with the possibility that the two protein isoforms encoded by the inebriated (ine) gene, which are members of the Na+/Cl--dependent neurotransmitter/osmolyte transporter family, perform osmolyte transport within the Malpighian tubule and hindgut. We show that ine mutants lacking both isoforms are hypersensitive to osmotic stress, which we assayed by maintaining flies on media containing NaCl, KCl, or sorbitol, and that this hypersensitivity is completely rescued by high-level ectopic expression of the ine-RB isoform. We provide evidence that this hypersensitivity represents a role for ine that is distinct from the increased neuronal excitability phenotype of ine mutants. Finally, we show that each ine genotype exhibits a "threshold" [NaCl]: long-term maintenance on NaCl-containing media above, but not below, the threshold causes lethality. Furthermore, this threshold value increases with the amount of ine activity. These data suggest that ine mutations confer osmotic stress sensitivity by preventing osmolyte accumulation within the Malpighian tubule and hindgut. PMID- 11861563 TI - Identification of circadian-clock-regulated enhancers and genes of Drosophila melanogaster by transposon mobilization and luciferase reporting of cyclical gene expression. AB - A new way was developed to isolate rhythmically expressed genes in Drosophila by modifying the classic enhancer-trap method. We constructed a P element containing sequences that encode firefly luciferase as a reporter for oscillating gene expression in live flies. After generation of 1176 autosomal insertion lines, bioluminescence screening revealed rhythmic reporter-gene activity in 6% of these strains. Rhythmically fluctuating reporter levels were shown to be altered by clock mutations in genes that specify various circadian transcription factors or repressors. Intriguingly, rhythmic luminescence in certain lines was affected by only a subset of the pacemaker mutations. By isolating genes near 13 of the transposon insertions and determining their temporal mRNA expression pattern, we found that four of the loci adjacent to the trapped enhancers are rhythmically expressed. Therefore, this approach is suitable for identifying genetic loci regulated by the circadian clock. One transposon insert caused a mutation in the rhythmically expressed gene numb. This novel numb allele, as well as previously described ones, was shown to affect the fly's rhythm of locomotor activity. In addition to its known role in cell fate determination, this gene and the phosphotyrosine-binding protein it encodes are likely to function in the circadian system. PMID- 11861564 TI - Interactions between natural selection, recombination and gene density in the genes of Drosophila. AB - In Drosophila, as in many organisms, natural selection leads to high levels of codon bias in genes that are highly expressed. Thus codon bias is an indicator of the intensity of one kind of selection that is experienced by genes and can be used to assess the impact of other genomic factors on natural selection. Among 13,000 genes in the Drosophila genome, codon bias has a slight positive, and strongly significant, association with recombination--as expected if recombination allows natural selection to act more efficiently when multiple linked sites segregate functional variation. The same reasoning leads to the expectation that the efficiency of selection, and thus average codon bias, should decline with gene density. However, this prediction is not confirmed. Levels of codon bias and gene expression are highest for those genes in an intermediate range of gene density, a pattern that may be the result of a tradeoff between the advantages for gene expression of close gene spacing and disadvantages arising from regulatory conflicts among tightly packed genes. These factors appear to overlay the more subtle effect of linkage among selected sites that gives rise to the association between recombination rate and codon bias. PMID- 11861565 TI - Recombinogenic effects of suppressors of position-effect variegation in Drosophila. AB - Compact chromatin structure, induction of gene silencing in position-effect variegation (PEV), and crossing-over suppression are typical features of heterochromatin. To identify genes affecting crossing-over suppression by heterochromatin we tested PEV suppressor mutations for their effects on crossing over in pericentromeric regions of Drosophila autosomes. From the 46 mutations (28 loci) studied, 16 Su(var) mutations of the nine genes Su(var)2-1, Su(var)2-2, Su(var)2-5, Su(var)2-10, Su(var)2-14, Su(var)2-15, Su(var)3-3, Su(var)3-7, and Su(var)3-9 significantly increase in heterozygotes or by additive effects in double and triple heterozygotes crossing over in the ri-p(p) region of chromosome 3. Su(var)2-2(01) and Su(var)2-14(01) display the strongest recombinogenic effects and were also shown to enhance recombination within the light-rolled heterochromatic region of chromosome 2. The dominant recombinogenic effects of Su(var) mutations are most pronounced in proximal euchromatin and are accompanied with significant reduction of meiotic nondisjunction. Our data suggest that crossing-over suppression by heterochromatin is controlled at chromatin structure as well as illustrate the possible effects of heterochromatin on total crossing over frequencies in the genome. PMID- 11861566 TI - Rough eye is a gain-of-function allele of amos that disrupts regulation of the proneural gene atonal during Drosophila retinal differentiation. AB - The regular organization of the ommatidial lattice in the Drosophila eye originates in the precise regulation of the proneural gene atonal (ato), which is responsible for the specification of the ommatidial founder cells R8. Here we show that Rough eye (Roi), a dominant mutation manifested by severe roughening of the adult eye surface, causes defects in ommatidial assembly and ommatidial spacing. The ommatidial spacing defect can be ascribed to the irregular distribution of R8 cells caused by a disruption of the patterning of ato expression. Disruptions in the recruitment of other photoreceptors and excess Hedgehog production in differentiating cells may further contribute to the defects in ommatidial assembly. Our molecular characterization of the Roi locus demonstrates that it is a gain-of-function mutation of the bHLH gene amos that results from a chromosomal inversion. We show that Roi can rescue the retinal developmental defect of ato1 mutants and speculate that amos substitutes for some of ato's function in the eye or activates a residual function of the ato1 allele. PMID- 11861567 TI - RNAi triggered by symmetrically transcribed transgenes in Drosophila melanogaster. AB - Specific silencing of target genes can be induced in a variety of organisms by providing homologous double-stranded RNA molecules. In vivo, these molecules can be generated either by transcription of sequences having an inverted-repeat (IR) configuration or by simultaneous transcription of sense-antisense strands. Since IR constructs are difficult to prepare and can stimulate genomic rearrangements, we investigated the silencing potential of symmetrically transcribed sequences. We report that Drosophila transgenes whose sense-antisense transcription was driven by two convergent arrays of Gal4-dependent UAS sequences can induce specific, dominant, and heritable repression of target genes. This effect is not dependent on a mechanism based on homology-dependent DNA/DNA interactions, but is directly triggered by transcriptional activation and is accompanied by specific depletion of the endogenous target RNA. Tissue-specific induction of these transgenes restricts the target gene silencing to selected body domains, and spreading phenomena described in other cases of post-transcriptional gene silencing (PTGS) were not observed. In addition to providing an additional tool useful for Drosophila functional genomic analysis, these results add further strength to the view that events of sense-antisense transcription may readily account for some, if not all, PTGS-cosuppression phenomena and can potentially play a relevant role in gene regulation. PMID- 11861568 TI - Sex-specific recombination rates in zebrafish (Danio rerio). AB - In many organisms, the rate of genetic recombination is not uniform along the length of chromosomes or between sexes. To compare the relative recombination rates during meiosis in male and female zebrafish, we constructed a genetic map based on male meiosis. We developed a meiotic mapping panel of 94 androgenetic haploid embryos that were scored for genetic polymorphisms. The resulting male map was compared to female and sex-average maps. We found that the recombination rate in male meiosis is dramatically suppressed relative to that of female meiosis, especially near the centromere. These findings have practical applications for experimental design. The use of exclusively female meiosis in a positional cloning project maximizes the ratio of genetic map distance to physical distance. Alternatively, the use of exclusively male meiosis to localize a mutation initially to a linkage group or to maintain relationships of linked alleles minimizes recombination, thereby facilitating some types of analysis. PMID- 11861569 TI - Molecular and phenotypic analysis of 25 recessive, homozygous-viable alleles at the mouse agouti locus. AB - Agouti is a paracrine-acting, transient antagonist of melanocortin 1 receptors that specifies the subapical band of yellow on otherwise black hairs of the wild type coat. To better understand both agouti structure/function and the germline damage caused by chemicals and radiation, an allelic series of 25 recessive, homozygous-viable agouti mutations generated in specific-locus tests were characterized. Visual inspection of fur, augmented by quantifiable chemical analysis of hair melanins, suggested four phenotypic categories (mild, moderate, umbrous-like, severe) for the 18 hypomorphs and a single category for the 7 amorphs (null). Molecular analysis indicated protein-coding alterations in 8 hypomorphs and 6 amorphs, with mild-moderate phenotypes correlating with signal peptide or basic domain mutations, and more devastating phenotypes resulting from C-terminal lesions. Ten hypomorphs and one null demonstrated wild-type coding potential, suggesting that they contain mutations elsewhere in the > or = 125-kb agouti locus that either reduce the level or alter the temporal/spatial distribution of agouti transcripts. Beyond the notable contributions to the field of mouse germ cell mutagenesis, analysis of this allelic series illustrates that complete abrogation of agouti function in vivo occurs most often through protein coding lesions, whereas partial loss of function occurs slightly more frequently at the level of gene expression control. PMID- 11861570 TI - A haplolethal locus uncovered by deletions in the mouse T complex. AB - Proper levels of gene expression are important for normal mammalian development. Typically, altered gene dosage caused by karyotypic abnormalities results in embryonic lethality or birth defects. Segmental aneuploidy can be compatible with life but often results in contiguous gene syndromes. The ability to manipulate the mouse genome allows the systematic exploration of regions that are affected by alterations in gene dosage. To explore the effects of segmental haploidy in the mouse t complex on chromosome 17, radiation-induced deletion complexes centered at the Sod2 and D17Leh94 loci were generated in embryonic stem (ES) cells. A small interval was identified that, when hemizygous, caused specific embryonic lethal phenotypes (exencephaly and edema) in most fetuses. The penetrance of these phenotypes was background dependent. Additionally, evidence for parent-of-origin effects was observed. This genetic approach should be useful for identifying genes that are imprinted or whose dosage is critical for normal embryonic development. PMID- 11861571 TI - Quantitative trait loci controlling light and hormone response in two accessions of Arabidopsis thaliana. AB - We have mapped quantitative trait loci (QTL) responsible for natural variation in light and hormone response between the Cape Verde Islands (Cvi) and Landsberg erecta (Ler) accessions of Arabidopsis thaliana using recombinant inbred lines (RILs). Hypocotyl length was measured in four light environments: white, blue, red, and far-red light and in the dark. In addition, white light plus gibberellin (GA) and dark plus the brassinosteroid biosynthesis inhibitor brassinazole (BRZ) were used to detect hormone effects. Twelve QTL were identified that map to loci not previously known to affect light response, as well as loci where candidate genes have been identified from known mutations. Some QTL act in all environments while others show genotype-by-environment interaction. A global threshold was established to identify a significant epistatic interaction between two loci that have few main effects of their own. LIGHT1, a major QTL, has been confirmed in a near isogenic line (NIL) and maps to a new locus with effects in all light environments. The erecta mutation can explain the effect of the HYP2 QTL in the blue, BRZ, and dark environments, but not in far-red. LIGHT2, also confirmed in an NIL, has effects in white and red light and shows interaction with GA. The phenotype and map position of LIGHT2 suggest the photoreceptor PHYB as a candidate gene. Natural variation in light and hormone response thus defines both new genes and known genes that control light response in wild accessions. PMID- 11861573 TI - Mapping of sex determination loci on the white campion (Silene latifolia) Y chromosome using amplified fragment length polymorphism. AB - S. latifolia is a dioecious plant with morphologically distinct sex chromosomes. To genetically map the sex determination loci on the male-specific Y chromosome, we identified X-ray-induced sex determination mutants that had lost male traits. We used male-specific AFLP markers to characterize the extent of deletions in the Y chromosomes of the mutants. We then compared overlapping deletions to predict the order of the AFLP markers and to locate the mutated sex-determining genes. We found three regions on the Y chromosome where frequent deletions were significantly associated with loss of male traits. One was associated with hermaphroditic mutants. A second was associated with asexual mutants that lack genes needed for early stamen development and a third was associated with asexual mutants that lack genes for late stages of stamen development. Our observations confirmed a classical genetic prediction that S. latifolia has three dispersed male-determining loci on the Y chromosome, one for carpel suppression, one for early stamen development, and another for late stamen development. This AFLP map provides a framework for locating genes on the Y chromosome and for characterizing deletions on the Y chromosomes of potentially interesting mutants. PMID- 11861572 TI - Maize Mu transposons are targeted to the 5' untranslated region of the gl8 gene and sequences flanking Mu target-site duplications exhibit nonrandom nucleotide composition throughout the genome. AB - The widespread use of the maize Mutator (Mu) system to generate mutants exploits the preference of Mu transposons to insert into genic regions. However, little is known about the specificity of Mu insertions within genes. Analysis of 79 independently isolated Mu-induced alleles at the gl8 locus established that at least 75 contain Mu insertions. Analysis of the terminal inverted repeats (TIRs) of the inserted transposons defined three new Mu transposons: Mu10, Mu 11, and Mu12. A large percentage (>80%) of the insertions are located in the 5' untranslated region (UTR) of the gl8 gene. Ten positions within the 5' UTR experienced multiple independent Mu insertions. Analyses of the nucleotide composition of the 9-bp TSD and the sequences directly flanking the TSD reveals that the nucleotide composition of Mu insertion sites differs dramatically from that of random DNA. In particular, the frequencies at which C's and G's are observed at positions -2 and +2 (relative to the TSD) are substantially higher than expected. Insertion sites of 315 RescueMu insertions displayed the same nonrandom nucleotide composition observed for the gl8-Mu alleles. Hence, this study provides strong evidence for the involvement of sequences flanking the TSD in Mu insertion-site selection. PMID- 11861574 TI - Assembly of two transgenes in an artificial chromatin domain gives highly coordinated expression in tobacco. AB - The chromatin loop model predicts that genes within the same chromatin domain exhibit coordinated regulation. We here present the first direct experimental support for this model in plants. Two reporter genes, the E. coli beta glucuronidase gene and the firefly luciferase gene, driven by different promoters, were placed between copies of the chicken lysozyme A element, a member of the matrix-associated region (MAR) group of chromatin boundary elements, and introduced in tobacco (Nicotiana tabacum). In plants carrying A elements, quantitative enzyme activities and mRNA levels of both genes show high correlations compared to control plants. The A element thus creates an artificial chromatin domain that yields coordinated expression. Surprisingly, enzyme activities correlated poorly with their respective mRNA levels. We hypothesize that this indicates the occurrence of "error pipelines" in data generation: systematic errors of a given analytical method will point in the same direction and cancel out in correlation analysis, resulting in better correlations. In combining different methods of analysis, however, such errors do not cancel out and as a result relevant correlations can be masked. Such error pipelines will have to be taken into account when different types of (e.g., whole-genome) data sets are combined in quantitative analyses. PMID- 11861575 TI - Likelihood-based estimation of the effective population size using temporal changes in allele frequencies: a genealogical approach. AB - A new genetic estimator of the effective population size (N(e)) is introduced. This likelihood-based (LB) estimator uses two temporally spaced genetic samples of individuals from a population. We compared its performance to that of the classical F-statistic-based N(e) estimator (N(eFk)) by using data from simulated populations with known N(e) and real populations. The new likelihood-based estimator (N(eLB)) showed narrower credible intervals and greater accuracy than (N(eFk)) when genetic drift was strong, but performed only slightly better when genetic drift was relatively weak. When drift was strong (e.g., N(e) = 20 for five generations), as few as approximately 10 loci (heterozygosity of 0.6; samples of 30 individuals) are sufficient to consistently achieve credible intervals with an upper limit <50 using the LB method. In contrast, approximately 20 loci are required for the same precision when using the classical F-statistic approach. The N(eLB) estimator is much improved over the classical method when there are many rare alleles. It will be especially useful in conservation biology because it less often overestimates N(e) than does N(eLB) and thus is less likely to erroneously suggest that a population is large and has a low extinction risk. PMID- 11861576 TI - A microsatellite-based multilocus screen for the identification of local selective sweeps. AB - With the availability of completely sequenced genomes, multilocus scans of natural variability have become a feasible approach for the identification of genomic regions subjected to natural and artificial selection. Here, I introduce a new multilocus test statistic, ln RV, which is based on the ratio of observed variances in repeat number at a set of microsatellite loci in two groups of populations. The distribution of ln RV values captures demographic history of the populations as well as variation in microsatellite mutation among loci. Given that microsatellite loci associated with a recent selective sweep differ from the remainder of the genome, they are expected to fall outside of the distribution of neutral ln RV values. The ln RV test statistic is applied to a data set of 94 loci typed in eight non-African and two African human populations. PMID- 11861577 TI - Detecting a local signature of genetic hitchhiking along a recombining chromosome. AB - The theory of genetic hitchhiking predicts that the level of genetic variation is greatly reduced at the site of strong directional selection and increases as the recombinational distance from the site of selection increases. This characteristic pattern can be used to detect recent directional selection on the basis of DNA polymorphism data. However, the large variance of nucleotide diversity in samples of moderate size imposes difficulties in detecting such patterns. We investigated the patterns of genetic variation along a recombining chromosome by constructing ancestral recombination graphs that are modified to incorporate the effect of genetic hitchhiking. A statistical method is proposed to test the significance of a local reduction of variation and a skew of the frequency spectrum caused by a hitchhiking event. This method also allows us to estimate the strength and the location of directional selection from DNA sequence data. PMID- 11861578 TI - Joint linkage and linkage disequilibrium mapping of quantitative trait loci in natural populations. AB - Linkage analysis and allelic association (also referred to as linkage disequilibrium) studies are two major approaches for mapping genes that control simple or complex traits in plants, animals, and humans. But these two approaches have limited utility when used alone, because they use only part of the information that is available for a mapping population. More recently, a new mapping strategy has been designed to integrate the advantages of linkage analysis and linkage disequilibrium analysis for genome mapping in outcrossing populations. The new strategy makes use of a random sample from a panmictic population and the open-pollinated progeny of the sample. In this article, we extend the new strategy to map quantitative trait loci (QTL), using molecular markers within the EM-implemented maximum-likelihood framework. The most significant advantage of this extension is that both linkage and linkage disequilibrium between a marker and QTL can be estimated simultaneously, thus increasing the efficiency and effectiveness of genome mapping for recalcitrant outcrossing species. Simulation studies are performed to test the statistical properties of the MLEs of genetic and genomic parameters including QTL allele frequency, QTL effects, QTL position, and the linkage disequilibrium of the QTL and a marker. The potential utility of our mapping strategy is discussed. PMID- 11861579 TI - Comparative analysis of the human dystrophin and utrophin gene structures. AB - We present analysis of intronic sequences in the human DMD and UTRN genes. In both genes accumulation of repeated elements could account for intron expansion. Out-of-frame rod-domain exons have stronger splice sites and are separated by significantly longer introns as compared to in-frame exons. These features are unique for the two homologs and not shared by other spectrin superfamily genes. PMID- 11861582 TI - The problem of children's injuries in low-income countries: a review. AB - Unintentional injuries are the cause of death and disability for millions of children every year in low-income countries. Challenging living conditions, heavy traffic, a lack of safe play space and an absence of child care options, together with a disproportionate vulnerability to injury, combine to put children at high risk. Inaccessible and unaffordable emergency services add to the number of resulting deaths and impairments. Yet this major public health problem receives relatively little attention. Because communicable disease and nutritional problems continue to rank higher as causes of child mortality and morbidity in most of the developing world, injury is perceived as a less serious problem. Existing research is scanty and is largely limited to hospital-based studies, which cannot present a comprehensive picture of either causes or outcomes. Development of preventive measures is hampered not only by limited health budgets, but by a tendency (not unique to low-income countries) to see injuries as random events, and hence as unpredictable and uncontrollable. There is an urgent need for more research that can contribute to effective analyses of the situation, and especially for locally-based research and record keeping, which is most likely to contribute to awareness and to practical and well-targeted prevention measures. PMID- 11861580 TI - Caenorhabditis elegans unc-37/groucho interacts genetically with components of the transcriptional mediator complex. AB - Groucho functions as a general corepressor by modulating chromatin structure and has a widespread role in many developmental processes. Here we show that Groucho may also interact with the basal transcriptional machinery. Mutations in Caenorhabditis elegans groucho interact with mutations in components of the transcriptional Mediator complex, resulting in synthetic lethality and loss of male sensory neurons. PMID- 11861581 TI - Multiple subunits of the Caenorhabditis elegans anaphase-promoting complex are required for chromosome segregation during meiosis I. AB - Two genes, originally identified in genetic screens for Caenorhabditis elegans mutants that arrest in metaphase of meiosis I, prove to encode subunits of the anaphase-promoting complex or cyclosome (APC/C). RNA interference studies reveal that these and other APC/C subunits are essential for the segregation of chromosomal homologs during meiosis I. Further, chromosome segregation during meiosis I requires APC/C functions in addition to the release of sister chromatid cohesion. PMID- 11861583 TI - Decentralization of health systems in Ghana, Zambia, Uganda and the Philippines: a comparative analysis of decision space. AB - This study reviews the experience of decentralization in four developing countries: Ghana, Uganda, Zambia and the Philippines. It uses two analytical frameworks to describe and compare the types and degrees of decentralization in each country. The first framework specifies three types of decentralization: deconcentration, delegation and devolution. The second framework uses a principal agent approach and innovative maps of 'decision space' to define the range of choice for different functions that is transferred from the centre to the periphery of the system. The analysis finds a variety of different types and degrees of decentralization, with the Philippines demonstrating the widest range of choice over many functions that were devolved to local government units. The least choice was transferred through delegation to an autonomous health service in Ghana. Uganda and Zambia display variations between these extremes. There was insufficient evidence of the impact of decentralization to assess how these differences in 'decision space' influenced the performance of each health system. The authors suggest that this is a major area for future research. PMID- 11861584 TI - Getting to the truth? Researching user views of primary health care. AB - In countries at all levels of development, assessing the opinions of health service users is increasingly promoted as an integral part of quality evaluations. However, there has been much debate on how best to measure user opinions. This article discusses findings from a study in South Africa, which employed both closed-ended facility exit interviews (total 337) and open-ended community-based focus group discussions (total 14) to obtain users' views on the same set of primary care providers. We outline various difficulties encountered in the interpretation of the data. First, in the absence of explicit and universal standards, users evaluated providers against their experiences with other health care services available to them in their areas. Responses were thus highly context specific, dependent on the particular configuration of services in each site. Secondly, the focus group discussions provided a very different (generally more negative) picture of providers to the exit interviews, suggesting that where and how views of health services are elicited has a large bearing on the results obtained. Thirdly, the focus group discussions appeared to encourage dramatic representations of what was, on observation, a banal everyday reality. Both methods defied superficial reading, and each appeared to have limitations in establishing the 'truth' about people's opinions. We conclude that there is a need for greater recognition, in quality assessments and in quality assurance, that user and community opinion is a social rather than a technical phenomenon. As such it is dynamic, bound to particular contexts and difficult to capture in single, 'snap-shot' assessments, no matter how well designed. In the context of quality assurance programmes, time spent assessing user views may be better used in other ways such as training and supporting health care workers to engage directly in dialogue with communities around needs and expectations. PMID- 11861585 TI - Perceived quality of care of primary health care services in Burkina Faso. AB - INTRODUCTION: Patients' views are being given more and more importance in policy making. Understanding populations' perceptions of quality of care is critical to developing measures to increase the utilization of primary health care services. OBJECTIVE: Documentation of user's opinion on the quality of care of primary health care services. METHODS: A 20-item scale, including four sub-scales related to health personnel practices and conduct, adequacy of resources and services, health care delivery, and financial and physical accessibility, was administered to 1081 users of 11 health care centres in the health district of Nouna, in rural Burkina Faso. RESULTS: The respondents were relatively positive on items related to health personnel practices and conduct and to health care delivery, but less so on items related to adequacy of resources and services and to financial and physical accessibility. In particular, the availability of drugs for all diseases on the spot, the adequacy of rooms and equipment in the facilities, the costs of care and the access to credit were valued poorly. Overall, the urban hospital was rated poorer than the average rural health care centre. Analysis of variance showed that, overall, health system characteristics explain 29% of all variation of the responses. CONCLUSION: Improving drug availability and financial accessibility to health services have been identified as the two main priorities for health policy action. Policy-makers should respect these patient preferences to deliver effective improvement of the quality of care as a potential means to increase utilization of health care. PMID- 11861586 TI - Primary health care vs. emergency medical assistance: a conceptual framework. AB - Primary health care (PHC) and emergency medical assistance (EMA) are discussed as two fundamentally different strategies of delivering health care. PHC is conceptualized as part of overall development, while EMA is delivered in disaster or emergency situations. The article contrasts the underlying paradigms, and the characteristics of care in PHC and EMA. It then analyzes the characteristics of PHC and EMA health services, their structure, management and support systems. In strategic aspects, it contrasts how managerial and financial sustainability are fundamentally different, and how the term accountability is used differently in development and disaster situations. However, while PHC and EMA, development and disaster, are clear opposite poles, many field situations in the developing world are today somewhere in-between. In such non-development, non-emergency situations, the objectives and approach will have to vary and an adapted strategy combining characteristics from PHC and EMA will have to be developed. PMID- 11861587 TI - Household utilization and expenditure on private and public health services in Vietnam. AB - The private provision of health services in Vietnam was legalized in 1989 as one of the country's means to mobilize resources and improve efficiency in the health system. Ten years after its legalization, the private sector has widely expanded its activities and become an important provider of health services for the Vietnamese people. However, little is known about its contribution to the overall objectives of the health system in Vietnam. This paper assesses the role of the private health care provider by examining utilization patterns and financial burden for households of private, as compared with public, services. We found that the private sector provided 60% of all outpatient contacts in Vietnam. There was no difference by education, sex or place of residence in the use of private ambulatory health care. Although there was evidence suggesting that rich people use private care more than the poor, this finding was not consistent across all income groups. The private sector served young children in particular. Also, people in households with several sick members at the same time relied more on private than public care, while those with severe illnesses tended to use less private care than public. The financial burden for households from private health care services was roughly a half of that imposed by the public providers. Expenditure on drugs accounted for a substantial percentage of household expenditure in general and health care expenditure in particular. These findings call for a prompt recognition of the private sector as a key player in Vietnam's health system. Health system policies should mobilize positive private sector contributions to health system goals where possible and reduce the negative effects of private provision development. PMID- 11861588 TI - Cost of access to health services in Zambia. AB - Equity is an important policy objective in the health care field. The importance of equity in health care provision can be argued from various points of view. As a result governments in all countries attempt to provide health care systems that enable equal access for everyone. Zambia is no exception. In the health care reforms the objective of the national health strategy is to provide Zambians with equity of access to health care. We focus on access defined as the costs (both monetary and time) an individual incurs when visiting a health care facility. Using a survey of 900 households, this article explores equality of access to health care among Zambians. Four areas are compared: urban high cost, urban low cost, townships and rural areas. The results of the analysis indicate that there are inequalities among residential areas, especially between rural and urban areas. In particular these differences exist because of differing distances to the nearest health facility. Large distances make it very costly for rural dwellers to seek medical care, especially during the high season for farming. The analysis suggests that obtaining equality of access to health care poses a challenge for the Zambian Government. PMID- 11861589 TI - Linking private and public sectors in tuberculosis treatment in Kathmandu Valley, Nepal. AB - Tuberculosis (TB) is a major public health problem and the world's foremost cause of death from a single infectious agent. Despite the increasing number of TB patients who seek help in the private sector, there are few practical examples of how to create a public/private linkage of TB services. The paper presents a pilot service-linkage project between the public and private sector in TB control in Kathmandu Valley, Nepal. The study documents and examines the process of the service-linkage project through the undertaking of a longitudinal analytical case study. A detailed description of the project from formulation to a short-term evaluation is given. The analysis relates the activities and early outcomes of the service-linkage project to the context, characteristics and interactions of the organizations involved. The study reveals that although the involved organizations initially agreed on the objective of the service-linkage project, differences in capacity, motivation, environment and needs had implications for the implementation and short-term success of the project. The public sector, despite the will, did not have the structure or resources to engage with the private sector. The private sector lacked interest in public health aspects of TB treatment and trust in the public sector. The study points to two different organizations that have the potential capacities to act as mediators between the public and private sectors: international research institutions and non governmental organizations. PMID- 11861590 TI - Delivery-related complications and determinants of caesarean section rates in India. AB - Caesarean section rates have been increasing worldwide, raising the question of the appropriateness of the selection of cases for the procedure. This paper examines the levels and correlates of delivery-related complications and caesarean section deliveries in 18 selected states of India in terms of specific maternal and institutional factors, using data from the National Family Health Surveys, 1992-93. Goa (15.3%) and Kerala (13.7%) were the two states with relatively higher caesarean section rates. There is reason to believe that current rates are part of a rising trend. This cannot be attributed entirely to the rise in institutional deliveries alone because of the strong association between caesarean sections and private sector institutions. Apart from the fact that the states of Kerala and Goa have relatively high caesarean section rates, in Andhra Pradesh, Bihar, Gujarat, Karnataka, Punjab and Uttar Pradesh the risk of undergoing caesarean section in private sector institutions is four or more times that in the public sector. It is possible that this extremely useful surgical procedure is being misused for profit purposes in the private sector in several states. There is therefore a need to examine this phenomenon using data disaggregated by the nature of caesarean sections, i.e. whether it was an elective or an emergency caesarean section along with the reasons for the choice. PMID- 11861591 TI - The sustainability of hepatitis B immunization within the Universal Immunization Programme in India. AB - Based on the recommendations of the World Health Organization, India as a member state is likely to implement universal immunization against hepatitis B through the existing Universal Immunization Programme (UIP). A pilot project is already under progress in two municipal zones of Delhi. This paper begins by reviewing epidemiological features of hepatitis B in India, some established aspects and other emerging trends. The gaps in the existing knowledge base are also given due consideration. The current recommendation is to deliver the vaccine at zero-day for infants in the absence of facilities for antenatal screening and immunoglobulin administration. The paper explores the potential pitfalls for integrating the proposed hepatitis B vaccination with the DPT (diphtheria/pertussis/tetanus) schedule. Based on the findings of the National Family Health Survey, the likely coverage for the states is estimated for both the schedules--zero-day and with DPT. The performance of the pilot phase is reviewed through the results of a coverage survey. The paper also estimates the resources that should be committed to launch the universal immunization of hepatitis B vaccination and the sustainability issues thereof. The paper finally concludes with the position that hepatitis B immunization will 'sink or sail' with the UIP. Further, this should act as an engine for recharging the infrastructure and functioning of the public health system and promote general preventive practices like universal precautions. PMID- 11861592 TI - Measuring social capital within health surveys: key issues. AB - With growing recognition of the social determinants of health, social capital is an increasingly important concept in international health research. Although there is relatively little experience of measuring social capital, particularly in developing countries, there are now a number of studies that allow the identification of some key issues that need to be considered when measuring social capital. After summarizing definitions and the rise of interest in the link between social capital and health, measures used in key studies of social capital and health are presented. Some important issues are then considered: obtaining a sufficiently comprehensive measure which empirically captures the latest theoretical developments in the field (particularly the concepts of cognitive, structural, bonding and bridging social capital); moving from the individual to the ecological level; consideration of confounding factors, and validity and reliability. PMID- 11861593 TI - Allowing for differential timing in cost analyses: discounting and annualization. AB - There are differences in timing related to when costs of certain inputs are incurred and when they are used over the lifetime of a programme. This paper looks at the issues related to the comparison of cost data over time focusing on discounting and annualization adjustments, which are used by economists to calculate financial and economic costs. The process of discounting is used to deal with the notion of time preference. Time preference implies that future costs are worth less, and hence discounted more, to reflect individual and societal preferences to have resources and money now rather than in the future. While discounting is appropriate in many situations, it is also useful to compute an annual equivalent cost when recurrent costs of an intervention are incurred, or are expected to be incurred, in subsequent years. This approach has the added benefit of illustrating how capital items are actually used during the lifetime of an intervention. This paper presents methods to both discount and annualize costs, and discusses rules-of-thumb to decide when to make these adjustments. PMID- 11861595 TI - A trip through my life with an immunological theme. AB - In this essay, I make four points about the operation of the immune system. First, thanks to the innate immune system's regulation of the main costimulatory molecules CD80 and CD86, the immune system rarely mistakes a pathogen for a self antigen. Second, the adaptive immune system consisting of T lymphocytes and B lymphocytes can mistake self for non-self because adaptive immunity is selected in single somatic cells. Third, the adaptive immune system of T lymphocytes and B lymphocytes is always referential to self, as it is selected on self-ligands; it persists in the periphery on self-ligands; and at least for T cells, it is dependent on self-ligands to be able to mount a response. Fourth, it is becoming clear that regulatory or suppressor T cells are our main defense against autoimmunity, as my first boss, Richard Gershon, had predicted. These cells recognize antigen as do all T cells, but they secrete the immunoregulatory cytokines IL-10 and TGF beta. PMID- 11861596 TI - The B7 family of ligands and its receptors: new pathways for costimulation and inhibition of immune responses. AB - T cell activation is dependent upon signals delivered through the antigen specific T cell receptor and accessory receptors on the T cell. A primary costimulatory signal is delivered through the CD28 receptor after engagement of its ligands, B7-1 (CD80) or B7-2 (CD86). Engagement of CTLA-4 (CD152) by the same B7-1 or B7-2 ligands results in attenuation of T cells responses. Recently, molecular homologs of CD28 and CTLA-4 receptors and their B7-like ligands have been identified. ICOS is a CD28-like costimulatory receptor with a unique B7-like ligand. PD-1 is an inhibitory receptor, with two B7-like ligands. Additional members of B7 and CD28 gene families have been proposed. Integration of signals through this family of costimulatory and inhibitory receptors and their ligands is critical for activation of immune responses and tolerance. Understanding these pathways will allow development of new strategies for therapeutic intervention in immune-mediated diseases. PMID- 11861597 TI - MAP kinases in the immune response. AB - MAP kinases are among the most ancient signal transduction pathways and are widely used throughout evolution in many physiological processes. In mammalian species, MAP kinases are involved in all aspects of immune responses, from the initiation phase of innate immunity, to activation of adaptive immunity, and to cell death when immune function is complete. In this review, we summarize recent progress in understanding the function and regulation of MAP kinase pathways in these phases of immune responses. PMID- 11861598 TI - Prospects for vaccine protection against HIV-1 infection and AIDS. AB - The rapid and devastating spread of the AIDS epidemic in the developing world as well as the difficulties associated with delivering antiretroviral drugs in affected countries underscore the urgent need for the development of a safe and effective AIDS vaccine. In this review, we discuss recent advances in our understanding of the cellular and humoral immune responses to human immunodeficiency virus type 1 (HIV-1) infection. We then describe vaccine strategies that have been explored and discuss the evidence suggesting that cellular immune responses elicited by novel vaccine modalities may attenuate clinical disease caused by HIV-1. PMID- 11861599 TI - T cell response in experimental autoimmune encephalomyelitis (EAE): role of self and cross-reactive antigens in shaping, tuning, and regulating the autopathogenic T cell repertoire. AB - T cells that can respond to self-antigens are present in the peripheral immune repertoire of all healthy individuals. Recently we have found that unmanipulated SJL mice that are highly susceptible to EAE also maintain a very high frequency of T cells responding to an encephalitogenic epitope of a myelin antigen proteolipid protein (PLP) 139-151 in the peripheral repertoire. This is not due to lack of expression of myelin antigens in the thymus resulting in escape of PLP 139-151 reactive cells from central tolerance, but is due to expression of a splice variant of PLP named DM20, which lacks the residues 116-150. In spite of this high frequency, the PLP 139-151 reactive cells remain undifferentiated in the periphery and do not induce spontaneous EAE. In contrast, SJL TCR transgenic mice expressing a receptor derived from a pathogenic T cell clone do develop spontaneous disease. This may be because in normal mice, autoreactive cells are kept in check by an alternate PLP 139-151 reactive nonpathogenic repertoire, which maintains a balance that keeps them healthy. If this is the case, selective activation of one repertoire or the other may alter susceptibility to autoimmune disease. Since T cells are generally cross-reactive, besides responding to nonself-antigens, they also maintain significant responses to self-antigens. Based on the PLP 139-151 system, we propose a model in which activation with foreign antigens can result in the generation of pathogenic memory T cells that mediate autoimmunity. We also outline circumstances under which activation of self-reactive T cells with foreign antigens can generate selective tolerance and thus generate protective/regulatory memory against self while still maintaining significant responses against foreign antigens. This provides a mechanism by which the fidelity and specificity of the immune system against foreign antigens is improved without increasing the potential for developing an autoimmune disease. PMID- 11861600 TI - Neuroendocrine regulation of immunity. AB - A reciprocal regulation exists between the central nervous and immune systems through which the CNS signals the immune system via hormonal and neuronal pathways and the immune system signals the CNS through cytokines. The primary hormonal pathway by which the CNS regulates the immune system is the hypothalamic pituitary-adrenal axis, through the hormones of the neuroendocrine stress response. The sympathetic nervous system regulates the function of the immune system primarily via adrenergic neurotransmitters released through neuronal routes. Neuroendocrine regulation of immune function is essential for survival during stress or infection and to modulate immune responses in inflammatory disease. Glucocorticoids are the main effector end point of this neuroendocrine system and, through the glucocorticoid receptor, have multiple effects on immune cells and molecules. This review focuses on the regulation of the immune response via the neuroendocrine system. Particular details are presented on the effects of interruptions of this regulatory loop at multiple levels in predisposition and expression of immune diseases and on mechanisms of glucocorticoid effects on immune cells and molecules. PMID- 11861601 TI - Molecular mechanism of class switch recombination: linkage with somatic hypermutation. AB - Class switch recombination (CSR) and somatic hypermutation (SHM) have been considered to be mediated by different molecular mechanisms because both target DNAs and DNA modification products are quite distinct. However, involvement of activation-induced cytidine deaminase (AID) in both CSR and SHM has revealed that the two genetic alteration mechanisms are surprisingly similar. Accumulating data led us to propose the following scenario: AID is likely to be an RNA editing enzyme that modifies an unknown pre-mRNA to generate mRNA encoding a nicking endonuclease specific to the stem-loop structure. Transcription of the S and V regions, which contain palindromic sequences, leads to transient denaturation, forming the stem-loop structure that is cleaved by the AID-regulated endonuclease. Cleaved single-strand tails will be processed by error-prone DNA polymerase-mediated gap-filling or exonuclease-mediated resection. Mismatched bases will be corrected or fixed by mismatch repair enzymes. CSR ends are then ligated by the NHEJ system while SHM nicks are repaired by another ligation system. PMID- 11861602 TI - Innate immune recognition. AB - The innate immune system is a universal and ancient form of host defense against infection. Innate immune recognition relies on a limited number of germline encoded receptors. These receptors evolved to recognize conserved products of microbial metabolism produced by microbial pathogens, but not by the host. Recognition of these molecular structures allows the immune system to distinguish infectious nonself from noninfectious self. Toll-like receptors play a major role in pathogen recognition and initiation of inflammatory and immune responses. Stimulation of Toll-like receptors by microbial products leads to the activation of signaling pathways that result in the induction of antimicrobial genes and inflammatory cytokines. In addition, stimulation of Toll-like receptors triggers dendritic cell maturation and results in the induction of costimulatory molecules and increased antigen-presenting capacity. Thus, microbial recognition by Toll like receptors helps to direct adaptive immune responses to antigens derived from microbial pathogens. PMID- 11861603 TI - KIR: diverse, rapidly evolving receptors of innate and adaptive immunity. AB - KIR genes have evolved in primates to generate a diverse family of receptors with unique structures that enable them to recognize MHC-class I molecules with locus and allele-specificity. Their combinatorial expression creates a repertoire of NK cells that surveys the expression of almost every MHC molecule independently, thus antagonizing the spread of pathogens and tumors that subvert innate and adaptive defense by selectively downregulating certain MHC class I molecules. The genes encoding KIR that recognize classical MHC molecules have diversified rapidly in human and primates; this contrasts with conservation of immunoglobulin and lectin-like receptors for nonclassical MHC molecules. As a result of the variable KIR-gene content in the genome and the polymorphism of the HLA system, dissimilar numbers and qualities of KIR:HLA pairs function in different humans. This diversity likely contributes variability to the function of NK cells and T lymphocytes by modulating innate and adaptive immune responses to specific challenges. PMID- 11861604 TI - Origins and functions of B-1 cells with notes on the role of CD5. AB - Whether B-1a (CD5+) cells are a distinct lineage derived from committed fetal/neonatal precursors or arise from follicular B-2 cells in response to BCR ligation and other, unknown signals remains controversial. Recent evidence indicates that B-1a cells can derive from adult precursors expressing an appropriate specificity when the (self-) antigen is present. Antibody specificity determines whether a B cell expressing immunoglobulin transgenes has a B-2, B-1a or marginal zone (MZ) phenotype. MZ cells share many phenotypic characteristics of B-1 cells and, like them, appear to develop in response to T independent type 2 antigens. Because fetal-derived B cell progenitors fail to express terminal deoxynucleotidyl transferase (TdT) and for other reasons, they are likely to express a repertoire that allows selection into the B-1a population. As it is selected by self-antigen, the B-1 repertoire tends to be autoreactive. This potentially dangerous repertoire is also useful, as B-1 cells are essential for resistance to several pathogens and they play an important role in mucosal immunity. The CD5 molecule can function as a negative regulator of BCR signaling that may help prevent inappropriate activation of autoreactive B-1a cells. PMID- 11861605 TI - E protein function in lymphocyte development. AB - Lymphocytes arise from hematopoietic stem cells through the coordinated action of transcription factors. The E proteins (E12, E47, HEB and E2-2) have emerged as key regulators of both B and T lymphocyte differentiation. This review summarizes the current data and examines the various functions of E proteins and their antagonists, Id2 and Id3, throughout lymphoid maturation. Beyond an established role in B and T lineage commitment, E proteins continue to be essential at subsequent stages of development. E protein activity regulates the expression of surrogate and antigen receptor genes, promotes Ig and TCR rearrangements, and coordinates cell survival and proliferation with developmental progression in response to TCR signaling. Finally, this review also discusses the role of E47 as a tumor suppressor. PMID- 11861606 TI - Lymphocyte-mediated cytotoxicity. AB - Virtually all of the measurable cell-mediated cytotoxicity delivered by cytotoxic T lymphocytes and natural killer cells comes from either the granule exocytosis pathway or the Fas pathway. The granule exocytosis pathway utilizes perforin to traffic the granzymes to appropriate locations in target cells, where they cleave critical substrates that initiate DNA fragmentation and apoptosis; granzymes A and B induce death via alternate, nonoverlapping pathways. The Fas/FasL system is responsible for activation-induced cell death but also plays an important role in lymphocyte-mediated killing under certain circumstances. The interplay between these two cytotoxic systems provides opportunities for therapeutic interventions to control autoimmune diseases and graft vs. host disease, but oversuppression of these pathways may also lead to increased viral susceptibility and/or decreased tumor cell killing. PMID- 11861607 TI - Signal transduction mediated by the T cell antigen receptor: the role of adapter proteins. AB - Engagement of the T cell antigen receptor (TCR) leads to a complex series of molecular changes at the plasma membrane, in the cytoplasm, and at the nucleus that lead ultimately to T cell effector function. Activation at the TCR of a set of protein tyrosine kinases (PTKs) is an early event in this process. This chapter reviews some of the critical substrates of these PTKs, the adapter proteins that, following phosphorylation on tyrosine residues, serve as binding sites for many of the critical effector enzymes and other adapter proteins required for T cell activation. The role of these adapters in binding various proteins, the interaction of adapters with plasma membrane microdomains, and the function of adapter proteins in control of the cytoskeleton are discussed. PMID- 11861608 TI - Interaction of heat shock proteins with peptides and antigen presenting cells: chaperoning of the innate and adaptive immune responses. AB - Heat shock proteins are abundant soluble intracellular proteins, present in all cells. Members of the heat shock protein family bind peptides including antigenic peptides generated within cells. Heat shock proteins also interact with antigen presenting cells through CD91 and other receptors, eliciting a cascade of events including re-presentation of heat shock protein-chaperoned peptides by MHC, translocation of NF kappa B into the nuclei and maturation of dendritic cells. These consequences point to a key role of heat shock proteins in fundamental immunological phenomena such as activation of antigen presenting cells, indirect presentation (or cross-priming), and chaperoning of peptides during antigen presentation. Heat shock proteins appear to have been involved in innate immune responses since the emergence of phagocytes in early multicellular organisms and to have been commandeered for adaptive immune responses with the advent of specificity. These properties of heat shock proteins also allow them to be used for immunotherapy of cancers and infections in novel ways. PMID- 11861609 TI - Chromatin structure and gene regulation in the immune system. AB - The development of the immune system and the host response to microbial infection rely on the activation and silencing of numerous, differentially expressed genes. Since the mid-1980s, a primary goal has been to identify transcription factors that regulate specific genes and specific immunological processes. More recently, there has been a growing appreciation of the role of chromatin structure in gene regulation. Before most activators of a gene access their binding sites, a transition from a condensed to a decondensed chromatin structure appears to take place. The activation of transcription is then accompanied by the remodeling of specific nucleosomes. Conversely, the acquisition of a more condensed chromatin structure is often associated with gene silencing. Chromatin structure is a particularly significant contributor to gene regulation because it is likely to be a major determinant of cell identity and cell memory. That is, the propagation of decondensed chromatin at specific loci through DNA replication and cell division helps a cell remember which genes are expressed constitutively in that cell type or are poised for expression upon exposure to a stimulus. Here we review recent progress toward understanding the role of chromatin in the immune system. The interleukin-4 gene serves as a primary model for exploring the events involved in the acquisition and heritable maintenance of a decondensed chromatin structure. Studies of the interleukin-12 p40 and interferon-beta genes are then reviewed for insight into the mechanisms by which the remodeling of specific nucleosomes in the vicinity of a promoter can contribute to rapid activation following cell stimulation. Finally, basic principles of gene silencing are discussed. PMID- 11861610 TI - Producing nature's gene-chips: the generation of peptides for display by MHC class I molecules. AB - Gene-chips contain thousands of nucleotide sequences that allow simultaneous analysis of the complex mixture of RNAs transcribed in cells. Like these gene chips, major histocompatibility complex (MHC) class I molecules display a large array of peptides on the cell surface for probing by the CD8(+) T cell repertoire. The peptide mixture represents fragments of most, if not all, intracellular proteins. The antigen processing machinery accomplishes the daunting task of sampling these proteins and cleaving them into the precise set of peptides displayed by MHC I molecules. It has long been believed that antigenic peptides arose as by-products of normal protein turnover. Recent evidence, however, suggests that the primary source of peptides is newly synthesized proteins that arise from conventional as well as cryptic translational reading frames. It is increasingly clear that for many peptides the C-terminus is generated in the cytoplasm, and N-terminal trimming occurs in the endoplasmic reticulum in an MHC I-dependent manner. Nature's gene-chips are thus both parsimonious and elegant. PMID- 11861611 TI - The immunology of mucosal models of inflammation. AB - In recent years the status of the inflammatory bowel diseases (IBDs) as canonical autoimmune diseases has risen steadily with the recognition that these diseases are, at their crux, abnormalities in mucosal responses to normally harmless antigens in the mucosal microflora and therefore responses to antigens that by their proximity and persistence are equivalent to self-antigens. This new paradigm is in no small measure traceable to the advent of multiple models of mucosal inflammation whose very existence is indicative of the fact that many types of immune imbalance can lead to loss of tolerance for mucosal antigens and thus inflammation centered in the gastrointestinal tract. We analyze the immunology of the IBDs through the lens of the murine models, first by drawing attention to their common features and then by considering individual models at a level of detail necessary to reveal their individual capacities to provide insight into IBD pathogenesis. What emerges is that murine models of mucosal inflammation have given us a road map that allows us to begin to define the immunology of the IBDs in all its complexity and to find unexpected ways to treat these diseases. PMID- 11861612 TI - T cell memory. AB - Typical immune responses lead to prominent clonal expansion of antigen-specific T and B cells followed by differentiation into effector cells. Most effector cells die at the end of the immune response but some of these cells survive and form long-lived memory cells. The factors controlling the formation and survival of memory T cells are reviewed. PMID- 11861613 TI - Genetic dissection of immunity to mycobacteria: the human model. AB - Humans are exposed to a variety of environmental mycobacteria (EM), and most children are inoculated with live Bacille Calmette-Guerin (BCG) vaccine. In addition, most of the world's population is occasionally exposed to human-borne mycobacterial species, which are less abundant but more virulent. Although rarely pathogenic, mildly virulent mycobacteria, including BCG and most EM, may cause a variety of clinical diseases. Mycobacterium tuberculosis, M. leprae, and EM M. ulcerans are more virulent, causing tuberculosis, leprosy, and Buruli ulcer, respectively. Remarkably, only a minority of individuals develop clinical disease, even if infected with virulent mycobacteria. The interindividual variability of clinical outcome is thought to result in part from variability in the human genes that control host defense. In this well-defined microbiological and clinical context, the principles of mouse immunology and the methods of human genetics can be combined to facilitate the genetic dissection of immunity to mycobacteria. The natural infections are unique to the human model, not being found in any of the animal models of experimental infection. We review current genetic knowledge concerning the simple and complex inheritance of predisposition to mycobacterial diseases in humans. Rare patients with Mendelian disorders have been found to be vulnerable to BCG, a few EM, and M. tuberculosis. Most cases of presumed Mendelian susceptibility to these and other mycobacterial species remain unexplained. In the general population leprosy and tuberculosis have been shown to be associated with certain human genetic polymorphisms and linked to certain chromosomal regions. The causal vulnerability genes themselves have yet to be identified and their pathogenic alleles immunologically validated. The studies carried out to date have been fruitful, initiating the genetic dissection of protective immunity against a variety of mycobacterial species in natural conditions of infection. The human model has potential uses beyond the study of mycobacterial infections and may well become a model of choice for the investigation of immunity to infectious agents. PMID- 11861614 TI - Antigen presentation and T cell stimulation by dendritic cells. AB - Dendritic cells take up antigens in peripheral tissues, process them into proteolytic peptides, and load these peptides onto major histocompatibility complex (MHC) class I and II molecules. Dendritic cells then migrate to secondary lymphoid organs and become competent to present antigens to T lymphocytes, thus initiating antigen-specific immune responses, or immunological tolerance. Antigen presentation in dendritic cells is finely regulated: antigen uptake, intracellular transport and degradation, and the traffic of MHC molecules are different in dendritic cells as compared to other antigen-presenting cells. These specializations account for dendritic cells' unique role in the initiation of immune responses and the induction of tolerance. PMID- 11861615 TI - Negative regulation of immunoreceptor signaling. AB - Immune cells are activated as a result of productive interactions between ligands and various receptors known as immunoreceptors. These receptors function by recruiting cytoplasmic protein tyrosine kinases, which trigger a unique phosphorylation signal leading to cell activation. In the recent past, there has been increasing interest in elucidating the processes involved in the negative regulation of immunoreceptor-mediated signal transduction. Evidence is accumulating that immunoreceptor signaling is inhibited by complex and highly regulated mechanisms that involve receptors, protein tyrosine kinases, protein tyrosine phosphatases, lipid phosphatases, ubiquitin ligases, and inhibitory adaptor molecules. Genetic evidence indicates that this inhibitory machinery is crucial for normal immune cell homeostasis. PMID- 11861616 TI - CpG motifs in bacterial DNA and their immune effects. AB - Unmethylated CpG motifs are prevalent in bacterial but not vertebrate genomic DNAs. Oligodeoxynucleotides (ODN) containing CpG motifs activate host defense mechanisms leading to innate and acquired immune responses. The recognition of CpG motifs requires Toll-like receptor (TLR) 9, which triggers alterations in cellular redox balance and the induction of cell signaling pathways including the mitogen activated protein kinases (MAPKs) and NF kappa B. Cells that express TLR 9, which include plasmacytoid dendritic cells (PDCs) and B cells, produce Th1 like proinflammatory cytokines, interferons, and chemokines. Certain CpG motifs (CpG-A) are especially potent at activating NK cells and inducing IFN-alpha production by PDCs, while other motifs (CpG-B) are especially potent B cell activators. CpG-induced activation of innate immunity protects against lethal challenge with a wide variety of pathogens, and has therapeutic activity in murine models of cancer and allergy. CpG ODN also enhance the development of acquired immune responses for prophylactic and therapeutic vaccination. PMID- 11861617 TI - Protein kinase C(theta) in T cell activation. AB - The novel protein kinase C (PKC) isoform, PKC theta, is selectively expressed in T lymphocytes and is a sine qua non for T cell antigen receptor (TCR)-triggered activation of mature T cells. Productive engagement of T cells by antigen presenting cells (APCs) results in recruitment of PKC theta to the T cell-APC contact area--the immunological synapse--where it interacts with several signaling molecules to induce activation signals essential for productive T cell activation and IL-2 production. The transcription factors NF-kappa B and AP-1 are the primary physiological targets of PKC theta, and efficient activation of these transcription factors by PKC theta requires integration of TCR and CD28 costimulatory signals. PKC theta cooperates with the protein Ser/Thr phosphatase, calcineurin, in transducing signals leading to activation of JNK, NFAT, and the IL-2 gene. PKC theta also promotes T cell cycle progression and regulates programmed T cell death. The exact mode of regulation and immediate downstream substrates of PKC theta are still largely unknown. Identification of these molecules and determination of their mode of operation with respect to the function of PKC theta will provide essential information on the mechanism of T cell activation. The selective expression of PKC theta in T cells and its essential role in mature T cell activation establish it as an attractive drug target for immunosuppression in transplantation and autoimmune diseases. PMID- 11861618 TI - RANK-L and RANK: T cells, bone loss, and mammalian evolution. AB - TNF and TNFR family proteins play important roles in the control of cell death, proliferation, autoimmunity, the function of immune cells, or the organogenesis of lymphoid organs. Recently, novel members of this large family have been identified that have critical functions in immunity and that couple lymphoid cells with other organ systems such as bone morphogenesis and mammary gland formation in pregnancy. The TNF-family molecule RANK-L (RANK-L, TRANCE, ODF) and its receptor RANK are key regulators of bone remodeling, and they are essential for the development and activation of osteoclasts. Intriguingly, RANK-L/RANK interactions also regulate T cell/dendritic cell communications, dendritic cell survival, and lymph node formation; T cell-derived RANK-L can mediate bone loss in arthritis and periodontal disease. Moreover, RANK-L and RANK are expressed in mammary gland epithelial cells, and they control the development of a lactating mammary gland during pregnancy and the propagation of mammalian species. Modulation of these systems provides us with a unique opportunity to design novel therapeutics to inhibit bone loss in arthritis, periodontal disease, and osteoporosis. PMID- 11861619 TI - Phagocytosis of microbes: complexity in action. AB - The phagocytic response of innate immune cells such as macrophages is defined by the activation of complex signaling networks that are stimulated by microbial contact. Many individual proteins have been demonstrated to participate in phagocytosis, and the application of high-throughput tools has indicated that many more remain to be described. In this review, we examine this complexity and describe how during recognition, multiple receptors are simultaneously engaged to mediate internalization, activate microbial killing, and induce the production of inflammatory cytokines and chemokines. Many signaling molecules perform multiple functions during phagocytosis, and these molecules are likely to be key regulators of the process. Indeed, pathogenic microorganisms target many of these molecules in their attempts to evade destruction. PMID- 11861620 TI - Structure and function of natural killer cell receptors: multiple molecular solutions to self, nonself discrimination. AB - In contrast to T cell receptors, signal transducing cell surface membrane molecules involved in the regulation of responses by cells of the innate immune system employ structures that are encoded in the genome rather than generated by somatic recombination and that recognize either classical MHC-I molecules or their structural relatives (such as MICA, RAE-1, or H-60). Considerable progress has recently been made in our understanding of molecular recognition by such molecules based on the determination of their three-dimensional structure, either in isolation or in complex with their MHC-I ligands. Those best studied are the receptors that are expressed on natural killer (NK) cells, but others are found on populations of T cells and other hematopoietic cells. These molecules fall into two major structural classes, those of the immunoglobulin superfamily (KIRs and LIRs) and of the C-type lectin-like family (Ly49, NKG2D, and CD94/NKG2). Here we summarize, in a functional context, the structures of the murine and human molecules that have recently been determined, with emphasis on how they bind different regions of their MHC-I ligands, and how this allows the discrimination of tumor or virus-infected cells from normal cells of the host. PMID- 11861621 TI - The informatics response in disaster, terrorism, and war. AB - The United States currently faces several new, concurrent large-scale health crises as a result of terrorist activity. In particular, three major health issues have risen sharply in urgency and public consciousness--bioterrorism, the threat of widespread delivery of agents of illness; mass disasters, local events that produce large numbers of casualties and overwhelm the usual capacity of health care delivery systems; and the delivery of optimal health care to remote military field sites. Each of these health issues carries large demands for the collection, analysis, coordination, and distribution of health information. The authors present overviews of these areas and discuss ongoing work efforts of experts in each. PMID- 11861622 TI - Roundtable on bioterrorism detection: information system-based surveillance. AB - During the 2001 AMIA Annual Symposium, the Anesthesia, Critical Care, and Emergency Medicine Working Group hosted the Roundtable on Bioterrorism Detection. Sixty-four people attended the roundtable discussion, during which several researchers discussed public health surveillance systems designed to enhance early detection of bioterrorism events. These systems make secondary use of existing clinical, laboratory, paramedical, and pharmacy data or facilitate electronic case reporting by clinicians. This paper combines case reports of six existing systems with discussion of some common techniques and approaches. The purpose of the roundtable discussion was to foster communication among researchers and promote progress by 1) sharing information about systems, including origins, current capabilities, stages of deployment, and architectures; 2) sharing lessons learned during the development and implementation of systems; and 3) exploring cooperation projects, including the sharing of software and data. A mailing list server for these ongoing efforts may be found at http://bt.cirg.washington.edu. PMID- 11861623 TI - The contributions of biomedical informatics to the fight against bioterrorism. AB - A comprehensive and timely response to current and future bioterrorist attacks requires a data acquisition, threat detection, and response infrastructure with unprecedented scope in time and space. Fortunately, biomedical informaticians have developed and implemented architectures, methodologies, and tools at the local and the regional levels that can be immediately pressed into service for the protection of our populations from these attacks. These unique contributions of the discipline of biomedical informatics are reviewed here. PMID- 11861624 TI - The space race and biodefense: lessons from NASA about big science and the role of medical informatics. AB - The events that followed the launch of Sputnik on Oct 4, 1957, provide a metaphor for the events that are following the first bioterroristic case of pulmonary anthrax in the United States. This paper uses that metaphor to elucidate the nature of the task ahead and to suggest questions such as, Can the goals of the biodefense effort be formulated as concisely and concretely as the goal of the space program? Can we measure success in biodefense as we did for the space project? What are the existing resources that are the equivalents of propulsion systems and rocket engineers that can be applied to the problems of biodefense? PMID- 11861625 TI - AMIA advocates national health information system in fight against national health threats. AB - To protect public health and national safety, AMIA recommends that the federal government dedicate technologic resources and medical informatics expertise to create a national health information infrastructure (NHII). An NHII provides the underlying information utility that connects local health providers and health officials through high-speed networks to national data systems necessary to detect and track global threats to public health. AMIA strongly recommends the accelerated development and wide-scale deployment of electronic public health surveillance systems, computer-based patient records, and disaster-response information technologies. Such efforts hold the greatest potential to protect our citizens from disaster and to deliver the best health care if disaster strikes. PMID- 11861628 TI - Training synergies between medical informatics and health services research: successes and challenges. AB - Stanford's two decades of success in linking medical informatics and health services research in both training and investigational activities reflects advantageous geography and history as well as natural synergies in the two areas. Health services research and medical informatics at Stanford have long shared a quantitative, analytic orientation, along with linked administration, curriculum, and clinical activities. Both the medical informatics and the health services research curricula draw on diverse course offerings throughout the university, and both the training and research overlap in such areas as outcomes research, large database analysis, and decision analysis/decision support. The Stanford experience suggests that successful integration of programs in medical informatics and health services research requires areas of overlapping or synergistic interest and activity among the involved faculty and, hence, in time, among the students. This is enhanced by a mixture of casual and structured contact among students from both disciplines, including social interactions. The challenges to integration are how to overcome any geographic separation that may exist in a given institution; the proper management of relationships with those sub-areas of medical informatics that have less overlap with health services research; and the need to determine how best to exploit opportunities for collaboration that naturally occur. PMID- 11861627 TI - Integrating medical informatics and health services research: the need for dual training at the clinical health systems and policy levels. AB - Reams of data pertaining directly to the core health services research mission are accumulating in large-scale organizational and clinical information systems. Health services researchers who grasp the structure of information systems and databases and the function of software applications can use existing data more effectively, assist in establishing new databases, and develop new tools to survey populations and collect data. At the same time, informaticians are needed who can structure databases that serve the needs of health service research and who can design and evaluate applications that effectively improve health care delivery. As long as health services researchers and informaticians work in separate spheres, however, opportunities to use data from health care encounters to improve care, expand knowledge, and develop more effective policies will be missed. This paper provides a brief exploration of 1) existing successful collaborations between health services researchers and informaticians and 2) needs and opportunities for additional joint work in several core research areas. PMID- 11861629 TI - Bridging the gap in medical informatics and health services research: workshop results and next steps. AB - In January 2000, the Agency for Healthcare Research and Quality (AHRQ) and the National Library of Medicine (NLM) cosponsored an invitational workshop entitled "Medical Informatics and Health Services Research: Bridging the Gap." Planned by a small committee of representatives from NLM and AHRQ institutional training centers, the workshop was designed to address the need for education of researchers interested in working at the intersection of the fields of medical informatics and health services research. More than 100 educators and researchers from AHRQ- and NLM-sponsored training programs in medical informatics and health services research participated in the workshop. Through a series of plenary presentations and breakout sessions, the workshop addressed ways of increasing the pool of persons interested, trained, and experienced in addressing specific areas of synergy between the two fields. This paper reports on the results of the workshop. PMID- 11861630 TI - Three decades of research on computer applications in health care: medical informatics support at the Agency for Healthcare Research and Quality. AB - The Agency for Healthcare Research and Quality and its predecessor organizations collectively referred to here as AHRQ-have a productive history of funding research and development in the field of medical informatics, with grant investments since 1968 totaling $107 million. Many computerized interventions that are commonplace today, such as drug interaction alerts, had their genesis in early AHRQ initiatives. This review provides a historical perspective on AHRQ investment in medical informatics research. It shows that grants provided by AHRQ resulted in achievements that include advancing automation in the clinical laboratory and radiology, assisting in technology development (computer languages, software, and hardware), evaluating the effectiveness of computer based medical information systems, facilitating the evolution of computer-aided decision making, promoting computer-initiated quality assurance programs, backing the formation and application of comprehensive data banks, enhancing the management of specific conditions such as HIV infection, and supporting health data coding and standards initiatives. Other federal agencies and private organizations have also supported research in medical informatics, some earlier and to a greater degree than AHRQ. The results and relative roles of these related efforts are beyond the scope of this review. PMID- 11861631 TI - Creating a pediatric digital library for pediatric health care providers and families: using literature and data to define common pediatric problems. AB - The goal of this study was to complete a literature-based needs assessment with regard to common pediatric problems encountered by pediatric health care providers (PHCPs) and families, and to develop a problem-based pediatric digital library to meet those needs. The needs assessment yielded 65 information sources. Common problems were identified and categorized, and the Internet was manually searched for authoritative Web sites. The created pediatric digital library (www.generalpediatrics.com) used a problem-based interface and was deployed in November 1999. From November 1999 to November 2000, the number of hyperlinks and authoritative Web sites increased 51.1 and 32.2 percent, respectively. Over the same time, visitors increased by 57.3 percent and overall usage increased by 255 percent. A pediatric digital library has been created that begins to bring order to general pediatric resources on the Internet. This pediatric digital library provides current, authoritative, easily accessed pediatric information whenever and wherever the PHCPs and families want assistance. PMID- 11861632 TI - Personal health records: evaluation of functionality and utility. AB - OBJECTIVES: Web-based applications have been developed that allow patients to enter their own information into secure personal health records. These applications are being promoted as a means of providing patients and providers with universal access to updated medical information. The authors evaluated the functionality and utility of a selection of personal health records. DESIGN: A targeted search strategy was used to identify eleven Web sites promoting different personal health records. Specific criteria related to the entry and display of data elements were developed to evaluate the functionality of each PHR. Information abstracted from an actual case was used to create a series of representative PHRs. Output generated for review was evaluated to assess the accuracy and completeness of clinical information related to the diagnosis and treatment of specific disorders. RESULTS: The PHRs selected for review employed data entry methods that limited the range and content of patient-entered information related to medical history, medications, laboratory tests, diagnostic studies, and immunizations. Representative PHRs created with information abstracted from an actual case displayed varying amounts of information at basic and comprehensive levels of representation. CONCLUSIONS: Currently available PHRs demonstrate limited functionality. The data entry, validation, and information display methods they employ may limit their utility as representations of medical information. PMID- 11861633 TI - Giving patients access to their medical records via the internet: the PCASSO experience. AB - OBJECTIVE: The Patient-Centered Access to Secure Systems Online (PCASSO) project is designed to apply state-of-the-art-security to the communication of clinical information over the Internet. DESIGN: The authors report the legal and regulatory issues associated with deploying the system, and results of its use by providers and patients. Human subject protection concerns raised by the Institutional Review Board focused on three areas-unauthorized access to information by persons other than the patient; the effect of startling or poorly understood information; and the effect of patient access to records on the record keeping behavior of providers. MEASUREMENTS: Objective and subjective measures of security and usability were obtained. RESULTS: During its initial deployment phase, the project enrolled 216 physicians and 41 patients; of these, 68 physicians and 26 patients used the system one or more times. The system performed as designed, with no unauthorized information access or intrusions detected. Providers rated the usability of the system low because of the complexity of the secure login and other security features and restrictions limiting their access to those patients with whom they had a professional relationship. In contrast, patients rated the usability and functionality of the system favorably. CONCLUSION: High-assurance systems that serve both patients and providers will need to address differing expectations regarding security and ease of use. PMID- 11861634 TI - Handheld technology to improve patient care: evaluating a support system for preference-based care planning at the bedside. AB - OBJECTIVE: Despite an increasing movement toward shared decision making and the incorporation of patients' preferences into health care decision making, little research has been done on the development and evaluation of support systems that help clinicians elicit and integrate patients' preferences into patient care. This study evaluates nurses' use of CHOICE, a handheld-computer-based support system for preference-based care planning, which assists nurses in eliciting patients' preferences for functional performance at the bedside. Specifically, it evaluates the effects of system use on nurses' care priorities, preference achievement, and patients' satisfaction. DESIGN: Three-group sequential design with one intervention and two control groups (N=155). In the intervention group, nurses elicited patients' preferences for functional performance with the handheld-computer-based CHOICE application as part of their regular admission interview; preference information was added to patients' charts and used in subsequent care planning. RESULTS: Nurses' use of CHOICE made nursing care more consistent with patient preferences (F=11.4; P<0.001) and improved patients' preference achievement (F=4.9; P<0.05). Furthermore, higher consistency between patients' preferences and nurses' care priorities was associated with higher preference achievement (r=0.49; P<0.001). CONCLUSION: In this study, the use of a handheld-computer-based support system for preference-based care planning improved patient-centered care and patient outcomes. The technique has potential to be included in clinical practice as part of nurses' routine care planning. PMID- 11861636 TI - AMIA recommendations for national health threat surveillance and response. PMID- 11861635 TI - Medical informatics and preparedness. PMID- 11861637 TI - Identification of the erythrocyte Rh blood group glycoprotein as a mammalian ammonium transporter. AB - The Rh blood group proteins are well known as the erythrocyte targets of the potent antibody response that causes hemolytic disease of the newborn. These proteins have been described in molecular detail; however, little is known about their function. A transport function is suggested by their predicted structure and from phylogenetic analysis. To obtain evidence for a role in solute transport, we expressed Rh proteins in Xenopus oocytes and now demonstrate that the erythroid Rh-associated glycoprotein mediates uptake of ammonium across cell membranes. Rh-associated glycoprotein carrier-mediated uptake, characterized with the radioactive analog of ammonium [(14)C]methylamine (MA), had an apparent EC(50) of 1.6 mm and a maximum uptake rate (V(max)) of 190 pmol/oocyte/min. Uptake was independent of the membrane potential and the Na(+) gradient. MA transport was stimulated by raising extracellular pH or by lowering intracellular pH, suggesting that uptake was coupled to an outwardly directed H(+) gradient. MA uptake was insensitive to additions of amiloride, amine-containing compounds tetramethyl- and tetraethylammonium chloride, glutamine, and urea. However, MA uptake was significantly antagonized by ammonium chloride with inhibition kinetics (IC(50) = 1.14 mm) consistent with the hypothesis that the uptake of MA and ammonium involves a similar H(+)-coupled counter-transport mechanism. PMID- 11861638 TI - Furin proteolytically processes the heparin-binding region of extracellular superoxide dismutase. AB - Extracellular superoxide dismutase (EC-SOD) is an antioxidant enzyme that attenuates brain and lung injury from oxidative stress. A polybasic region in the carboxyl terminus distinguishes EC-SOD from other superoxide dismutases and determines EC-SOD's tissue half-life and affinity for heparin. There are two types of EC-SOD that differ based on the presence or absence of this heparin binding region. It has recently been shown that proteolytic removal of the heparin-binding region is an intracellular event (Enghild, J. J., Thogersen, I. B., Oury, T. D., Valnickova, Z., Hojrup, P., and Crapo, J. D. (1999) J. Biol. Chem. 274, 14818-14822). By using mammalian cell lines, we have now determined that removal of the heparin-binding region occurs after passage through the Golgi network but before being secreted into the extracellular space. Specific protease inhibitors and overexpression of intracellular proteases implicate furin as a processing protease. In vitro experiments using furin and purified EC-SOD suggest that furin proteolytically cleaves EC-SOD in the middle of the polybasic region and then requires an additional carboxypeptidase to remove the remaining lysines and arginines. A mutation in Arg(213) renders EC-SOD resistant to furin processing. These results indicate that furin-dependent processing of EC-SOD is important for determining the tissue distribution and half-life of EC-SOD. PMID- 11861640 TI - Enhancement of hyperthermia-induced apoptosis by local anesthetics on human histiocytic lymphoma U937 cells. AB - The combined effects of hyperthermia at 44 degrees C and local anesthetics on apoptosis in human histiocytic lymphoma U937 cells were investigated. When the cells were exposed to hyperthermia for l0 min marginal DNA fragmentation and nuclear fragmentation were observed. In the presence of amide-type local anesthetics further enhancement was found depending on concentration. The order of the concentration required for maximum induction was the reverse order of the lipophilicity (prilocaine > lidocaine > bupivacaine). Western blotting revealed that in hyperthermia there was initial release of Ca(2+) from the intracellular store site as indicated by increased expression of the type 1 inositol-1,4,5 trisphosphate receptor. However, the combination with lidocaine did not induce any further enhancement. Lidocaine enhanced the decrease in ATP content and the increase in intracellular Ca(2+) concentration in individual cells induced by hyperthermia. In addition, superoxide formation, decrease in the mitochondrial membrane potential, and activation of intracellular caspase-3 were found in the cells treated with hyperthermia and lidocaine. All of these were suppressed in part in the presence of the intracellular Ca(2+) ion chelator BAPTA-AM (bis-(O aminophenoxy)-ethane-N,N,N',N'-tetraacetic acid-acetoxymethyl). The present results indicate that local anesthetics at optimal concentrations enhance hyperthermia-induced apoptosis via Ca(2+)- and mitochondria-dependent pathways. Initial release of Ca(2+) from intracellular store sites caused by hyperthermia and followed by the subsequent increase in the intracellular Ca(2+) concentration and the additional activation of the mitochondrial caspase-dependent pathway (partly regulated by intracellular Ca(2+) concentration) plays a crucial role in the enhancement of apoptosis induced by the combination of hyperthermia and lidocaine. PMID- 11861639 TI - Mertk triggers uptake of photoreceptor outer segments during phagocytosis by cultured retinal pigment epithelial cells. AB - The RCS rat is a widely studied model of recessively inherited retinal degeneration. The genetic defect, known as rdy (retinal dystrophy), results in failure of the retinal pigment epithelium (RPE) to phagocytize shed photoreceptor outer segment membranes. We previously used positional cloning and in vivo genetic complementation to demonstrate that Mertk is the gene for rdy. We have now used a rat primary RPE cell culture system to demonstrate that the RPE is the site of action of Mertk and to obtain functional evidence for a key role of Mertk in RPE phagocytosis. We found that Mertk protein is absent from RCS, but not wild type, tissues and cultured RPE cells. Delivery of rat Mertk to cultured RCS RPE cells by means of a recombinant adenovirus restored the cells to complete phagocytic competency. Infected RCS RPE cells ingested exogenous outer segments to the same extent as wild-type RPE cells, but outer segment binding was unaffected. Mertk protein progressively co-localized with outer segment material during phagocytosis by primary RPE cells, and activated Mertk accumulated during the early stages of phagocytosis by RPE-J cells. We conclude that Mertk likely functions directly in the RPE phagocytic process as a signaling molecule triggering outer segment ingestion. PMID- 11861641 TI - Activation of UBC5 ubiquitin-conjugating enzyme by the RING finger of ROC1 and assembly of active ubiquitin ligases by all cullins. AB - Protein ubiquitination plays an important role in regulating the abundance and conformation of a broad range of eukaryotic proteins. This process involves a cascade of enzymes including ubiquitin-activating enzymes (E1), ubiquitin conjugating enzymes (E2), and ubiquitin ligases (E3). E1 and E2 represent two families of structurally related proteins and are relatively well characterized. In contrast, the nature and mechanism of E3, proposed to contain activities in catalyzing isopeptide bond formation (ubiquitin ligation) and substrate targeting, remains inadequately understood. Two major families of E3 ubiquitin ligases, the HECT (for homologous to E6-AP C terminus) family and the RING family, have been identified that utilize distinct mechanisms in promoting isopeptide bond formation. Here, we showed that purified RING finger domain of ROC1, an essential subunit of SKP1-cullin/CDC53-F box protein ubiquitin ligases, was sufficient to activate UBCH5c to synthesize polyubiquitin chains. The sequence flanking the RING finger in ROC1 did not contribute to UBCH5c activation, but was required for binding with CUL1. We demonstrated that all cullins, through their binding with ROC proteins, constituted active ubiquitin ligases, suggesting the existence in vivo of a large number of cullin-RING ubiquitin ligases. These results are consistent with the notion that the RING finger domains allosterically activate E2. We suggest that RING-E2, rather than cullin-RING, constitutes the catalytic core of the ubiquitin ligase and that one major function of the cullin subunit is to assemble the RING-E2 catalytic core and substrates together. PMID- 11861642 TI - Specific modification of mitochondrial protein thiols in response to oxidative stress: a proteomics approach. AB - Mitochondria play a central role in redox-linked processes in the cell through mechanisms that are thought to involve modification of specific protein thiols, but this has proved difficult to assess. In particular, specific labeling and quantitation of mitochondrial protein cysteine residues have not been achieved due to the lack of reagents available that can be applied to the intact organelle or cell. To overcome these problems we have used a combination of mitochondrial proteomics and targeted labeling of mitochondrial thiols using a novel compound, (4-iodobutyl)triphenylphosphonium (IBTP). This lipophilic cation is accumulated by mitochondria and yields stable thioether adducts in a thiol-specific reaction. The selective uptake into mitochondria, due to the large membrane potential across the inner membrane, and the high pH of the matrix results in specific labeling of mitochondrial protein thiols by IBTP. Individual mitochondrial proteins that changed thiol redox state following oxidative stress could then be identified by their decreased reaction with IBTP and isolated by two-dimensional electrophoresis. We demonstrate the selectivity of IBTP labeling and use it to show that glutathione oxidation and exposure to an S-nitrosothiol or to peroxynitrite cause extensive redox changes to mitochondrial thiol proteins. In conjunction with blue native gel electrophoresis, we used IBTP labeling to demonstrate that thiols are exposed on the matrix faces of respiratory Complexes I, II, and IV. This novel approach enables measurement of the thiol redox state of individual mitochondrial proteins during oxidative stress and cell death. In addition the methodology has the potential to identify novel redox-dependent modulation of mitochondrial proteins. PMID- 11861644 TI - A novel function of BCL-2 overexpression in regulatory volume decrease. Enhancing swelling-activated Ca(2+) entry and Cl(-) channel activity. AB - The cellular function of the oncogene bcl-2, a key regulator of apoptosis, is still debated. The goal of this study was to explore the relationship between BCL 2 overexpression and cell volume regulation by using two independent models, Madin-Darby canine kidney (MDCK) cells stably transfected with BCL-2 and MDCK clones with inducible BCL-2 expression by the lac operator/repressor. BCL-2 overexpression enhanced the capability of regulatory volume decrease (RVD), a cellular defensive process against hypotonic stress. In various clones of MDCK cells, hypotonic stress induced an outwardly rectified Cl(-) current that was significantly up-regulated by BCL-2 overexpression. Other fundamental characteristics of this channel were similar among different MDCK clones, such as sensitivity to Cl(-) channel inhibitor, anion permeability, and time-dependent inactivation at more positive potential. Most importantly, BCL-2 overexpression up-regulates the swelling-activated Ca(2+) transient that is a critical signaling for normal RVD and the activation of swelling-activated Cl(-) channel in MDCK cells. BCL-2 overexpression also enhances the capacitative Ca(2+) entry that can be differentiated from the swelling-activated Ca(2+) transient by kinetic analysis and sensitivity to Gd(3+). Moreover, neutralization of endogenous BCL-2 by antibody blocks the normal RVD response and the activation of swelling activated Cl(-) channel in human cervical cancer HT-3 cells. These results provide a new insight into the novel function of BCL-2 overexpression in the regulation of cell volume and ion flux. PMID- 11861645 TI - Hypoxia sensitizes cells to nitric oxide-induced apoptosis. AB - Nitric oxide (NO) can induce apoptosis in a variety of cell types. A non-toxic concentration of nitric oxide under normal oxygen conditions triggered cell death under hypoxic conditions (1.5% O(2)) in fibroblasts. Nitric oxide administered during hypoxia induced the release of cytochrome c, caspase-9 activation, and the loss of mitochondrial membrane potential followed by DNA fragmentation and lactate dehydrogenase release (markers of cell death). Bcl-X(L) protected cells from nitric oxide-induced apoptosis during hypoxia by preventing the release of cytochrome c, caspase-9 activation, and by maintaining a mitochondrial membrane potential. Murine embryonic fibroblasts from bax(-/-) bak(-/-) mice exposed to nitric oxide during hypoxia did not die, indicating that pro-apoptotic Bcl-2 family members are required for NO-induced apoptosis during hypoxia. The nitric oxide-induced cell death during hypoxia was independent of cGMP and peroxynitrite. Cells devoid of mitochondrial DNA (rho secondary-cells) lack a functional electron transport chain and were resistant to nitric oxide-induced cell death during hypoxia, suggesting that a functional electron transport chain is required for nitric oxide-induced apoptosis during hypoxia. PMID- 11861643 TI - Analysis of conserved active site residues in monoamine oxidase A and B and their three-dimensional molecular modeling. AB - Monoamine oxidase (MAO) is a key enzyme responsible for the degradation of serotonin, norepinephrine, dopamine, and phenylethylamine. It is an outer membrane mitochondrial enzyme existing in two isoforms, A and B. We have recently generated 14 site-directed mutants of human MAO A and B, and we found that four key amino acids, Lys-305, Trp-397, Tyr-407, and Tyr-444, in MAO A and their corresponding amino acids in MAO B, Lys-296, Trp-388, Tyr-398, and Tyr-435, play important roles in MAO catalytic activity. Based on the polyamine oxidase three dimensional crystal structure, it is suggested that Lys-305, Trp-397, and Tyr-407 in MAO A and Lys-296, Trp-388, and Tyr-398 in MAO B may be involved in the non covalent binding to FAD. Tyr-407 and Tyr-444 in MAO A (Tyr-398 and Tyr-435 in MAO B) may form an aromatic sandwich that stabilizes the substrate binding. Asp-132 in MAO A (Asp-123 in MAO B) located at the entrance of the U-shaped substrate binding site has no effect on MAO A nor MAO B catalytic activity. The similar impact of analogous mutants in MAO A and MAO B suggests that these amino acids have the same function in both isoenzymes. Three-dimensional modeling of MAO A and B using polyamine oxidase as template suggests that the overall tertiary structure and the active sites of MAO A and B may be similar. PMID- 11861646 TI - The First Nucleotide Binding Domain of Cystic Fibrosis Transmembrane Conductance Regulator Is a Site of Stable Nucleotide Interaction, whereas the Second Is a Site of Rapid Turnover. AB - As in other adenine nucleotide binding cassette (ABC) proteins the nucleotide binding domains of the cystic fibrosis transmembrane conductance regulator (CFTR) bind and hydrolyze ATP and in some manner regulate CFTR ion channel gating. Unlike some other ABC proteins, however, there are preliminary indications that the two domains of CFTR are nonequivalent in their nucleotide interactions (Szabo, K., Szakacs, G., Hegeds, T., and Sarkadi, B. (1999) J. Biol. Chem. 274, 12209-12212; Aleksandrov, L., Mengos, A., Chang, X., Aleksandrov, A., and Riordan, J. R. (2001) J. Biol. Chem. 276, 12918-12923). We have now characterized the interactions of the 8-azido-photoactive analogues of ATP, ADP, and 5'-adenyl beta,gamma-imidodiphosphate (AMP-PNP) with the two domains of functional membrane bound CFTR. The results show that the two domains appear to act independently in the binding and hydrolysis of 8-azido-ATP. At NBD1 binding does not require a divalent cation. This binding is followed by minimal Mg(2+)-dependent hydrolysis and retention of the hydrolysis product, 8-azido-ADP, but not as a vanadate stabilized post-hydrolysis transition state complex. In contrast, at NBD2, MgN(3)ATP is hydrolyzed as rapidly as it is bound and the nucleoside diphosphate hydrolysis product dissociates immediately. Confirming this characterization of NBD1 as a site of more stable nucleotide interaction and NBD2 as a site of fast turnover, the non-hydrolyzable N(3)AMP-PNP bound preferentially to NBD1. This demonstration of NBD2 as the rapid nucleotide turnover site is consistent with the strong effect on channel gating kinetics of inactivation of this domain by mutagenesis. PMID- 11861647 TI - Glycogen synthase kinase-3 beta mutagenesis identifies a common binding domain for GBP and Axin. AB - Glycogen synthase kinase-3 beta (GSK-3) is a key downstream target of Wnt signaling and is regulated by its interactions with activating and inhibitory proteins. We and others have shown that GSK-3 activity toward non-primed substrates is regulated in part through a competition between its activating (Axin) and inhibitory (GBP/FRAT) binding partners. Here we use a reverse two hybrid screen to identify mutations in GSK-3 that alter binding to GBP and Axin. We find that these mutations overlap and propose that GBP and Axin compete for binding to the same region of GSK-3. We use these mutations to examine the ability of GSK-3 to block eye development in Xenopus embryos and suggest that GSK 3 regulates eye development through a non-Wnt pathway. PMID- 11861649 TI - Accurate translation of the genetic code depends on tRNA modified nucleosides. AB - Transfer RNA molecules translate the genetic code by recognizing cognate mRNA codons during protein synthesis. The anticodon wobble at position 34 and the nucleotide immediately 3' to the anticodon triplet at position 37 display a large diversity of modified nucleosides in the tRNAs of all organisms. We show that tRNA species translating 2-fold degenerate codons require a modified U(34) to enable recognition of their cognate codons ending in A or G but restrict reading of noncognate or near-cognate codons ending in U and C that specify a different amino acid. In particular, the nucleoside modifications 2-thiouridine at position 34 (s(2)U(34)), 5-methylaminomethyluridine at position 34 (mnm(5)U(34)), and 6 threonylcarbamoyladenosine at position 37 (t(6)A(37)) were essential for Watson Crick (AAA) and wobble (AAG) cognate codon recognition by tRNA(UUU)(Lys) at the ribosomal aminoacyl and peptidyl sites but did not enable the recognition of the asparagine codons (AAU and AAC). We conclude that modified nucleosides evolved to modulate an anticodon domain structure necessary for many tRNA species to accurately translate the genetic code. PMID- 11861648 TI - A mouse serine protease TESP5 is selectively included into lipid rafts of sperm membrane presumably as a glycosylphosphatidylinositol-anchored protein. AB - We have previously indicated that at least in mouse, sperm serine protease(s) other than acrosin probably act on the limited proteolysis of egg zona pellucida to create a penetration pathway for motile sperm, although the participation of acrosin cannot be ruled out completely. A 42-kDa gelatin-hydrolyzing serine protease present in mouse sperm is a candidate enzyme involved in the sperm penetration of the zona pellucida. In this study, we have PCR-amplified an EST clone encoding a testicular serine protease, termed TESP5, and then screened a mouse genomic DNA library using the DNA fragment as a probe. The DNA sequence of the isolated genomic clones indicated that the TESP5 gene is identical to the genes coding for testicular testisin and eosinophilic esp-1. Immunochemical analysis using affinity-purified anti-TESP5 antibody revealed that 42- and 41-kDa forms of TESP5 with the isoelectric points of 5.0 to 5.5 are localized in the head, cytoplasmic droplet, and midpiece of cauda epididymal sperm probably as a membranous protein. Moreover, these two forms of TESP5 were selectively included into Triton X-100-insoluble microdomains, lipid rafts, of the sperm membranes. These results show the identity between TESP5/testisin/esp-1 and the 42-kDa sperm serine protease. When HEK293 cells were transformed by an expression plasmid carrying the entire protein-coding region of TESP5, the recombinant protein produced was released from the cell membrane by treatment with Bacillus cereus phosphatidylinositol-specific phospholipase C, indicating that TESP5 is glycosylphosphatidylinositol-anchored on the cell surface. Enzymatic properties of recombinant TESP5 was similar to but distinguished from those of rat acrosin and pancreatic trypsin by the substrate specificity and inhibitory effects of serine protease inhibitors. PMID- 11861650 TI - Differential role of the proline-rich domain of nuclear factor 1-C splice variants in DNA binding and transactivation. AB - We have addressed the functional significance of the existence of several natural splice variants of NF1-C* differing in their COOH-terminal proline-rich transactivation domain (PRD) by studying their specific DNA binding and transactivation in the yeast Saccharomyces cerevisiae. These parameters yielded the intrinsic transactivation potential (ITP), defined as the activation observed with equal amounts of DNA bound protein. Exchange of 83 amino acids at the COOH terminal end of the PRD by 16 unrelated amino acids, as found in NF1-C2, and splicing out the central region of the PRD, as found in NF1-C7, enhanced DNA binding in vivo and in vitro. However, the ITP of the splice variants NF1-C2 and NF1-C7 was found to be similar to that of the intact NF1-C1. Additional mutations showed that the ITP of NF1-C requires the synergistic action of the PRD and a novel domain encoded in exons 5 and 6. Intriguingly the carboxyl-terminal domain like motif encoded in exons 9/10 is not essential for transactivation of a reporter with a single NF1 site but is required for activation of a reporter with six NF1 sites in tandem. Our results imply that differential splicing is used to regulate transcription by generating variants with different DNA binding affinities but similar ITPs. PMID- 11861651 TI - mu-Opioid receptors desensitize less rapidly than delta-opioid receptors due to less efficient activation of arrestin. AB - Receptor desensitization by G-protein receptor kinases (GRK) and arrestins is likely to be an important component underlying the development of tolerance to opioid drugs. Reconstitution of this process in Xenopus oocytes revealed distinct differences in the kinetics of GRK and arrestin regulation of the closely related opioid receptors mu (MOR), delta (DOR), and kappa (KOR). We demonstrated that under identical conditions, GRK and arrestin-dependent desensitization of MOR proceeds dramatically slower than that of DOR. Furthermore, GRK3 phosphorylation sites required for opioid receptor desensitization also greatly differ. The determinants for DOR and KOR desensitization reside in the carboxyl-terminal tail, whereas MOR depends on Thr-180 in the second intracellular loop. Although this later finding might indicate an inefficient phosphorylation of MOR Thr-180, increasing the amount of arrestin expressed greatly increased the rate of MOR desensitization to a rate comparable with that of DOR. Similarly, coexpression of a constitutively active arrestin 2(R169E) with MOR and DOR desensitized both receptors in an agonist-dependent, GRK-independent manner at rates that were indistinguishable. Together, these data suggest that it is the activation of arrestin, rather than its binding, that is the rate-limiting step in MOR desensitization. In addition, mutation of Thr-161 in DOR, homologous to MOR Thr 180, significantly inhibited the faster desensitization of DOR. These results suggest that DOR desensitization involves phosphorylation of both the carboxyl terminal tail and the second intracellular loop that together leads to a more efficient activation of arrestin and thus faster desensitization. PMID- 11861652 TI - Reconstituted discoidal ApoE-phospholipid particles are ligands for the scavenger receptor BI. The amino-terminal 1-165 domain of ApoE suffices for receptor binding. AB - The high density lipoprotein receptor, scavenger receptor class B type I (SR-BI), recognizes lipid-bound apolipoprotein A-I (apoA-I) and other apolipoproteins. Here, we have used large scale cultures of apoE-expressing cells to purify apoE and prepare apoE containing reconstituted discoidal 1-palmitoyl-2-oleoyl-l phosphatidylcholine (POPC)-apoE particles. These particles have been used to examine their binding to wild-type and mutant forms of SR-BI expressed in transfected ldlA-7 cells. Specific binding to SR-BI was determined by subtracting from the total binding, nonspecific values measured using either control untransfected ldlA-7 cells or by inhibiting SR-BI-mediated binding with a high titer antireceptor-blocking antibody. POPC-apoE particles generated using apoE2, apoE3, apoE4, or the carboxyl-terminally truncated forms apoE165, apoE202, apoE229, and apoE259 all bound tightly to wild-type SR-BI with similar affinities (K(d) = 35-45 microg/ml). Binding was nearly abolished in a cell line expressing the ldlA (Q402R/Q418R) double mutant form of SR-BI that is unable to bind native high density lipoprotein but binds low density lipoprotein normally. The findings establish that apoE is a ligand for SR-BI and that the receptor binding domain is located in the amino-terminal 1-165-region of the protein. SR-BI-apoE interactions may contribute to cholesterol homeostasis in tissues and cells expressing SR-BI that are accessible to apoE-containing lipoproteins. PMID- 11861653 TI - Unfolding and double-stranded DNA binding of the cold shock protein homologue Cla h 8 from Cladosporium herbarum. AB - The cloning, purification, and biophysical characterization of the first eukaryotic cold shock protein homologue, Cla h 8, expressed as single functional polypeptide is reported here. It was discovered as a minor allergen of the mold Cladosporium herbarum by phage display using a library selectively enriched for IgE-binding proteins. Based on the sequence homology of Cla h 8 with bacterial cold shock proteins (CSPs), a homology-based computer model of the allergen was computed indicating an all-beta structure of Cla h 8. This major structural feature was confirmed by CD spectroscopy. Despite the structural similarities with bacterial CSPs, the DNA-binding and unfolding behavior of Cla h 8 exhibited unique and previously undescribed characteristics. High affinities of Cla h 8 for single-stranded DNA as well as for double-stranded DNA corresponding to the human Y-box were detected. The affinity for double-stranded DNA increased significantly with decreasing temperature, which was paralleled by an increase in the beta sheet content of the protein. Temperature-dependent fluorescence anisotropy and far-UV CD measurements revealed different unfolding transitions at 28 and at 35.7 degrees C, respectively, indicating a multistate transition, which is uncommon for CSPs. The enhanced affinity for DNA at low temperatures together with the low unfolding transition refer to the functional significance of Cla h 8 at reduced temperatures. PMID- 11861654 TI - GD3 recruits reactive oxygen species to induce cell proliferation and apoptosis in human aortic smooth muscle cells. AB - Sialic acid containing glycosphingolipids (gangliosides) are expressed on the surface of all mammalian cells and have been implicated in regulating various biological phenomena; however, the detailed signaling mechanisms involved in this process are not known. We report here a novel aspect of disialoganglioside, GD3 mediated regulation of cell proliferation and cell death via the recruitment of reactive oxygen species (ROS). A low concentration (2.5-10 microm) of GD3, incubated with human aortic smooth muscle cells for a short period of time (10-30 min), stimulates superoxide generation via the activation of both NADPH oxidase and NADH oxidase activity. This leads to downstream signaling leading to cell proliferation and apoptosis. However, [(3)H]GD3 incubated with the cells under such conditions was found in a trypsin-sensitive fraction that was separable from endogenous GD3. The exact mechanism causing ROS generation and downstream signaling remains to be elucidated. The uptake of GD3 was accompanied by a 2.5 fold stimulation in the activity of mitogen-activated protein (MAP) kinase and 5 fold stimulation in cell proliferation. Preincubation of cells with membrane permeable antioxidants, pyrrolidine dithiocarbamate, and N-acetylcysteine abrogated the superoxide generation and cell proliferation. In contrast, at higher concentrations (50-200 microm) GD3 inhibited the generation of superoxides but markedly stimulated the generation of nitric oxide (NO) (10-fold compared with control). This in turn stimulated mitochondrial cytochrome c release and intrachromosomal DNA fragmentation, which lead to apoptosis. In sum, at a low concentration, GD3 recruits superoxides to activate p44 MAPK and stimulates cell proliferation. In contrast, at high concentrations GD3 recruits nitric oxide to scavenge superoxide radicals that triggered signaling events that led to apoptosis. These observations might have relevance in regard to the potential role of GD3 in aortic smooth muscle cell proliferation and apoptosis that may contribute to plaque rupture in atherosclerosis. PMID- 11861655 TI - Intracellular amyloid-beta 1-42, but not extracellular soluble amyloid-beta peptides, induces neuronal apoptosis. AB - Alzheimer disease (AD), the most frequent cause of dementia, is characterized by an important neuronal loss. A typical histological hallmark of AD is the extracellular deposition of beta-amyloid peptide (A beta), which is produced by the cleavage of the amyloid precursor protein (APP). Most of the gene mutations that segregate with the inherited forms of AD result in increasing the ratio of A beta 42/A beta 40 production. A beta 42 also accumulates in neurons of AD patients. Altogether, these data strongly suggest that the neuronal production of A beta 42 is a critical event in AD, but the intraneuronal A beta 42 toxicity has never been demonstrated. Here, we report that the long term expression of human APP in rat cortical neurons induces apoptosis. Although APP processing leads to production of extracellular A beta 1-40 and soluble APP, these extracellular derivatives do not induce neuronal death. On the contrary, neurons undergo apoptosis as soon as they accumulate intracellular A beta 1-42 following the expression of full-length APP or a C-terminal deleted APP isoform. The inhibition of intraneuronal A beta 1-42 production by a functional gamma-secretase inhibitor increases neuronal survival. Therefore, the accumulation of intraneuronal A beta 1-42 is the key event in the neurodegenerative process that we observed. PMID- 11861656 TI - Macrophage-restricted and interferon gamma-inducible expression of the allograft inflammatory factor-1 gene requires Pu.1. AB - Expression of allograft inflammatory factor-1 (Aif-1), a 17-kDa protein bearing an EF-hand Ca(2+) binding motif, increases markedly in monocytes and macrophages participating in allo- and autoimmune reactions, including the perivascular inflammation in transplanted hearts, microglial infiltrates in experimental autoimmune neuritis, and the inflamed pancreas of prediabetic BB rats. To investigate the mechanism of this regulation, we isolated the mouse aif-1 gene and determined its genomic organization. The gene has six exons distributed over 1.6 kilobases, an interferon gamma-inducible DNase I-hypersensitive site near 900, and flanking sequences on either side predicted to associate with nuclear matrix. Reporter gene analyses identified sequences between -902 and -789, including consensus Ets and interferon regulatory factor elements, required for macrophage-specific and interferon gamma-inducible transcriptional activity. Pu.1 bound to the Ets site in electromobility shift assay and forced expression of Pu.1 activated the aif-1 promoter in 3T3 fibroblasts, in which it is normally inactive. However, the transcriptional activity of a concatamer of the Ets site alone did not increase with interferon gamma treatment. Cooperation between Pu.1 and proteins binding to the interferon regulatory factor element appears to be necessary for both macrophage-specific and interferon gamma-inducible expression of the aif-1 gene. PMID- 11861657 TI - Ca2+ uptake and release properties of a thapsigargin-insensitive nonmitochondrial Ca2+ store in A7r5 and 16HBE14o- cells. AB - In a previous study we overexpressed the thapsigargin (tg)-insensitive Pmr1 Ca(2+) pump of the Golgi apparatus of Caenorhabditis elegans in COS-1 cells and studied the properties of the Ca(2+) store into which it was integrated. Here we assessed the properties of an endogenous tg-insensitive nonmitochondrial Ca(2+) store in A7r5 and 16HBE14o- cells, which express a mammalian homologue of Pmr1. The tg-insensitive Ca(2+) store was considerably less leaky for Ca(2+) than the sarco(endo)plasmic-reticulum Ca(2+)-ATPase (SERCA)-containing Ca(2+) store. Moreover like for the worm Pmr1 Ca(2+) pump expressed in COS-1 cells, Ca(2+) accumulation into the endogenous tg-insensitive store showed a 2 orders of magnitude lower sensitivity to cyclopiazonic acid than the SERCA-mediated transport. 2,5-Di-(tert-butyl)-1,4-benzohydroquinone was only a very weak inhibitor of the tg-insensitive Ca(2+) uptake in A7r5 and 16HBE14o- cells and in COS-1 cells overexpressing the worm Pmr1. Inositol 1,4,5-trisphosphate released 11% of the Ca(2+) accumulated in permeabilized A7r5 cells pretreated with tg with an EC(50) that was 5 times higher than for the SERCA-containing Ca(2+) store but failed to release Ca(2+) in 16HBE14o- cells. In the presence of tg, 15% of intact A7r5 cells responded to 10 microm arginine-vasopressin with a small rise in cytosolic Ca(2+) concentration after a long latency. In conclusion, A7r5 and 16HBE14o- cells express a Pmr1-containing Ca(2+) store with properties that differ substantially from the SERCA-containing Ca(2+) store. PMID- 11861659 TI - PPARadigms and PPARadoxes: expanding roles for PPARgamma in the control of lipid metabolism. AB - The nuclear receptor PPARgamma is a central regulator of adipose tissue development and an important modulator of gene expression in a number of specialized cell types including adipocytes, epithelial cells, and macrophages. PPARgamma signaling pathways impact both cellular and systemic lipid metabolism and have links to obesity, diabetes, and cardiovascular disease. The ability to activate this receptor with small molecule ligands has made PPARgamma an attractive target for intervention in human metabolic disease. As our understanding of PPARgamma biology has expanded, so has the therapeutic potential of PPARgamma ligands. Recent studies have provided insight into the paradoxical relationship between PPARgamma and metabolic disease and established new paradigms for the control of lipid metabolism. This review focuses on recent advances in PPARgamma biology in the areas of adipocyte differentiation, insulin resistance, and atherosclerosis. PMID- 11861660 TI - Interaction of apolipoprotein A-I in three different conformations with palmitoyl oleoyl phosphatidylcholine vesicles. AB - Interactions of apolipoprotein A-I (apoA-I) with cell membranes appear to be important in the initial steps of reverse cholesterol transport. The objective of this work was to examine the effect of three distinct conformations of apoA-I (lipid-free and in 78 A or 96 A reconstituted high density lipoproteins, rHDL) on its ability to bind to, and abstract lipids from, palmitoyl oleoyl phosphatidylcholine membrane vesicles (small unilamellar vesicles, SUV, and giant unilamellar vesicles, GUV). The molecular interactions were observed by two photon fluorescence microscopy, and the binding parameters were quantified by gel permeation chromatography or isothermal titration microcalorimetry. Rearrangement of apoA-I-containing particles after exposure to SUVs was examined by native gel electrophoresis. The results indicate that lipid-free apoA-I binds reversibly, with high affinity, to the vesicles but does not abstract a significant amount of lipid nor perturb the vesicle structure. The 96 A rHDL, where all the amphipathic helices of apoA-I are saturated with lipid within the particles, do not bind to vesicles or perturb their structure. In contrast, the 78 A rHDL have a region of apoA-I, corresponding to a few amphipathic helical segments, which is available for external or internal phospholipid binding. These particles bind to vesicles with measurable affinity (lower than lipid-free apoA-I), abstract lipids from the membranes, and form particles of larger diameters, including 96 A rHDL. We conclude that the conformation of apoA-I regulates its binding affinity for phospholipid membranes and its ability to abstract lipids from the membranes. PMID- 11861661 TI - Dynamics of lysosomal cholesterol in Niemann-Pick type C and normal human fibroblasts. AB - The dynamics of endolysosomal cholesterol were investigated in Niemann-Pick type C (NPC) cells and in human fibroblasts treated with class 2 amphiphiles to mimic NPC cells. We showed through new approaches that the massive pools of endolysosomal cholesterol in these cells are not trapped but, rather, circulate to the cell surface at about the normal rate. This flux spared NPC and amphiphile treated cells from disruption by the extraction of their plasma membrane cholesterol with cyclodextrin. Nocodazole, a microtubule-depolymerizing agent, reversed the resistance of NPC and U18666A-treated cells to cholesterol depletion, apparently by reducing the flux of endolysosomal cholesterol to the plasma membrane. Neither nocodazole nor bafilomycin A1 (an inhibitor of the vacuolar proton pump) acted in the same way as the NPC mutation or class 2 amphiphiles: both agents decreased plasma membrane cholesterol at the expense of the endolysosomal pool and both blocked the actions of the amphiphile, U18666A. Finally, the resistance of NPC cells to lysis by amphotericin B was shown not to reflect a reduction in plasma membrane cholesterol arising from a block in lysosomal cholesterol export but rather the diversion of the amphotericin B to cholesterol-rich endolysosomes. We conclude that the large pool of endolysosomal cholesterol in NPC and amphiphile-treated fibroblasts is dynamic and that its turnover, as in normal cells, is dependent on microtubules. PMID- 11861662 TI - Increased glycosphingolipid levels in serum and aortae of apolipoprotein E gene knockout mice. AB - The apolipoprotein E gene knockout (apoE-/-) mouse develops atherosclerosis that shares many features of human atherosclerosis. Increased levels of glycosphingolipid (GSL) have been reported in human atherosclerotic lesions; however, GSL levels have not been studied in the apoE-/- mouse. Here we used HPLC methods to analyze serum and aortic GSL levels in apoE-/- and C57BL/6J control mice. The concentrations of glucosyl ceramide (GlcCer), lactosyl ceramide (LacCer), GalNAcbeta1-4Galbeta1-4Glc-Cer (GA2), and ceramide trihexoside (CTH) were increased by approximately 7-fold in the apoE-/- mouse serum compared with controls. The major serum ganglioside, N-glycolyl GalNAcbeta1-4[NeuNAcalpha2 3]Galbeta1-4Glc-Cer (N-glycolyl GM2), was increased in concentration by approximately 3-fold. A redistribution of GSLs from HDL to VLDL populations was also observed in the apoE-/- mice. These changes were accompanied by an increase in the levels of GSLs in the aortic sinus and arch of the apoE-/- mice. The spectrum of gangliosides present in the aortic tissues was more complex than that found in the lipoproteins, with the latter represented almost entirely by N glycolyl GM2 and the former comprised of NeuNAcalpha2-3Galbeta1-4Glc-Cer (GM3), GM2, N-glycolyl GM2, GM1, GD3, and GD1a. In conclusion, neutral GSL and ganglioside levels were increased in the serum and aortae of apoE-/- mice compared with controls, and this was associated with a preferential redistribution of GSL to the proatherogenic lipoprotein populations. The apoE-/- mouse therefore represents a useful model to study the potential role of GSL metabolism in atherogenesis. PMID- 11861663 TI - Macrophage lipoprotein lipase expression is increased in patients with heterozygous familial hypercholesterolemia. AB - FH is associated with accelerated atherosclerosis. Based on the crucial role of macrophage LPL in atherogenesis, we determined in the present study macrophage LPL expression in patients with FH. Monocytes isolated from 13 FH patients and 13 control subjects were differentiated into macrophages by culturing the cells for 9 days in 20% autologous or heterologous serum. Macrophages of patients with FH cultured in their own sera showed a significant increase in LPL mRNA levels, extracellular LPL mass, and activity compared with macrophages of control subjects. Although these alterations positively correlated with the levels of serum platelet-derived growth factor-BB (PDGF-BB) in FH subjects, increased LPL secretion by cultured FH macrophages was reduced neither by immunoneutralizing FH serum with an anti-PDGF-BB antibody, nor by culturing these cells in sera from control subjects. With the exception of LPL, levels of other cytokines and 8 isoprostane were not increased in the supernatants of macrophages of FH patients. Serum from FH patients also enhances the levels of LPL secreted by macrophages from control subjects. Immunoneutralization of FH serum with an anti-PDGF-BB antibody totally reversed this alteration. Overall, this study demonstrates that macrophages from FH subjects overproduce LPL and that PDGF present in the serum from FH patients stimulates LPL secretion by control macrophages. These findings suggest that macrophage LPL induction in patients with FH might be related to the increased atherogenesis observed in these subjects. PMID- 11861664 TI - Use of stable isotopically labeled tracers to measure very low density lipoprotein-triglyceride turnover. AB - Tracer methods for VLDL-TG kinetics vary in their ability to account for the effect of tracer recycling, which can influence the calculation of VLDL-TG fractional catabolic rates (FCRs). We evaluated a novel approach, involving stable isotopically labeled glycerol or palmitate tracers in conjunction with compartmental modeling, for measuring VLDL-TG kinetics in normolipidemic human subjects. When administered as a bolus simultaneously, both tracers provided identical VLDL-TG FCRs when the data were analyzed by a compartmental model that accounted for hepatic lipid tracer recycling, but not by non-compartmental analysis. The model-derived FCR was greater than that determined using a non compartmental approach, and was 2- to 3-fold higher than that usually reported by using a bolus of radioactive [3H]glycerol. When palmitate tracer was given as a constant infusion, VLDL-TG turnover appeared 5-fold slower, because tracer recycling through hepatic lipid pools could not be resolved with the infusion protocol. We conclude that accounting for tracer recycling, particularly the contribution of hepatic glycerolipid pools, is essential to accurately measure VLDL-TG kinetics, and that bolus injection of stable isotopically labeled glycerol or palmitate tracers in conjunction with compartmental modeling analysis offers a reliable approach for measuring VLDL-TG kinetics. PMID- 11861665 TI - Apolipoprotein [a] genotype influences isoform dominance pattern differently in African Americans and Caucasians. AB - Plasma lipoprotein [a] (Lp[a]) concentrations are inversely associated with, and largely determined by, apolipoprotein [a] (apo[a]) gene size, a highly polymorphic trait. We studied if, within an individual, the smaller apo[a] isoform always dominated, whether there was interaction between the two alleles, and whether these features differed between Caucasians and African Americans. We determined apo[a] gene sizes, apo[a] protein sizes and relative amounts, and plasma Lp[a] levels in 430 individuals (263 Caucasians and 167 African Americans). Of the 397 heterozygotes with at least one detectable apo[a] isoform (238 Caucasians and 159 African Americans), the larger allele dominated in 28% of Caucasians and 23% of African Americans, while the smaller allele dominated in 56% of Caucasians and 45% of African Americans. In Caucasians, dominance of the smaller allele increased with Lp[a] levels, from 44% at Lp[a] < or = 30 nM to 81% at Lp[a] >100 nM (P < 0.0001). Dominance by the smaller allele increased with increasing size of the larger allele in both groups but with the smaller allele only in African Americans. There was no interaction between apo[a] alleles within genotypes; one apo[a] isoform level was not associated with the other isoform level, and isoform levels were not affected by the difference in size. More of the dominance pattern was explained by Lp[a] level and apo[a] genotype in African Americans than in Caucasians (29% vs. 13%). Thus, genotype influences isoform specific Lp[a] levels and dominance patterns differently in African Americans and in Caucasians. PMID- 11861666 TI - ORP2, a homolog of oxysterol binding protein, regulates cellular cholesterol metabolism. AB - Oxysterol binding protein (OSBP) related proteins (ORPs) constitute a family that has at least 12 members in humans. In the present study we characterize one of the novel OSBP homologs, ORP2, which we show to be expressed ubiquitously in mammalian tissues. The ORP2 cDNA encodes a deduced 55 kDa protein that lacks a pleckstrin homology (PH) domain, a feature found in the other family members. Sucrose gradient centrifugation analysis of Chinese hamster ovary (CHO) cell post nuclear supernatant demonstrated that ORP2 is distributed in soluble and membrane bound fractions. Immunofluorescence microscopy of the endogenous and overexpressed ORP2 in CHO cells suggested that the membrane-bound fraction of the protein localizes to the Golgi apparatus. Stably transfected CHO cells that overexpress ORP2 showed an increase in [14C]cholesterol efflux to serum, apolipoprotein A-I (apoA-I), and phosphatidyl choline vesicles. The proportion of cellular [14C]cholesterol that is esterified and the ACAT activity measured as [14C]oleyl-CoA conversion into cholesteryl [14C]oleate by the cellular membranes, were markedly decreased in the ORP2 expressing cells. Transient high level overexpression of ORP2 interfered with the clearance of a secretory pathway protein marker from the Golgi complex. The results implicate ORP2 as a novel regulator of cellular sterol homeostasis and intracellular membrane trafficking. PMID- 11861667 TI - Altered distribution of platelet-activating factor- acetylhydrolase activity between LDL and HDL as a function of the severity of hypercholesterolemia. AB - Platelet-activating factor-acetylhydrolase (PAF-AH) is a lipoprotein-associated phospholipase A2 capable of hydrolyzing platelet-activating factor (PAF) and oxidatively modified phospholipids. We studied the plasma- and lipoprotein associated PAF-AH activity in patients with primary hypercholesterolemia. Thirty eight unrelated patients with heterozygous familial hypercholesterolemia (HeteroFH), five patients with homozygous FH (HomoFH), and 33 patients with primary non-FH hypercholesterolemia (NonFH) participated in the study. In all patient groups the plasma PAF-AH activity was significantly elevated compared with 33 normolipidemic controls, the HomoFH having the highest and the NonFH patients showing the lowest enzyme activity. Gradient ultracentrifugation studies showed that this increase is not only due to the elevation in the plasma LDL but also to the increase in the PAF-AH activity associated with each LDL subfraction, being more profound in the small-dense LDL-5. Unlike LDL, no difference in the HDL-associated PAF-AH activity was observed among all groups. Consequently, an altered distribution of enzyme activity among apolipoprotein B (apoB)- and apolipoprotein A-I (apoA-I)-containing lipoproteins is observed in hypercholesterolemic patients, resulting in a significant decrease in the ratio of the HDL-associated PAF-AH to the total plasma enzyme activity compared with controls. This reduction is proportional to the increase of the plasma LDL cholesterol (LDL-C) levels and consequently to the severity of the hypercholesterolemia. Thus, the ratio of HDL-associated PAF-AH-total plasma enzyme activity may be useful as a potential marker of atherogenicity in subjects with primary hypercholesterolemia. PMID- 11861668 TI - Efficacy of a Therapeutic Lifestyle Change/Step 2 diet in moderately hypercholesterolemic middle-aged and elderly female and male subjects. AB - Lifestyle modification to decrease cardiovascular disease (CVD) risk has recently been reaffirmed by both the National Cholesterol Education Program and American Heart Association (AHA). Using a randomized crossover design, the Therapeutic Lifestyle Change (TLC)/Step 2 diet relative to a typical Western diet was assessed in 36 moderately hypercholesterolemic subjects in a clinical setting under isoweight conditions. Mean lipoprotein and apolipoprotein levels (fasting and non-fasting), fatty acid profiles, parameters of HDL metabolism, and glucose homeostasis were determined. Relative to the Western diet, the TLC/Step 2 diet resulted in 11% and 7% lower LDL cholesterol (LDL-C) and HDL cholesterol (HDL-C), respectively, with no significant change in TG levels or total cholesterol-HDL-C ratio. Similar responses were observed in the non-fasting state. Linoleic (18:2n 6c) and alpha-linolenic (18:3n-3) acids increased at the expense of oleic acid (18:1n-9c) in the cholesteryl ester, TG, and phospholipid subfractions. The dietary changes had no significant effect on fractional esterification rate of HDL, phospholipid transfer protein (PLTP), or cholesterol ester transfer protein activities, or glucose and insulin levels. Female and male subjects responded similarly. The TLC/Step 2 diet resulted in a decrease in some CVD risk factors and no apparent adverse effects in others. PMID- 11861669 TI - Postprandial changes of apoB-100 and apoB-48 in TG rich lipoproteins in familial combined hyperlipidemia. AB - Impaired chylomicron (chylo) remnant clearance and small VLDL overproduction are major metabolic abnormalities in familial combined hyperlipidemia (FCHL). Quantitative data on postprandial apolipoprotein B-48 (apoB-48) and apolipoprotein B-100 (apoB-100) in TG rich lipoproteins (TRL) in FCHL have not been reported before. Eight untreated FCHL patients and 10 matched controls underwent a 24 h oral fat load. Fasting apoB-48 and apoB-100 were significantly higher in all TRL in FCHL. Maximal concentrations of chylo-[Svedberg's flotation rate (Sf) >400] apoB-48 and apoB-100 were reached later in FCHL (at t = 6 h), in contrast to controls (t = 4 h). Maximal VLDL1-(Sf60-400)-apoB-48 occurred at the same time point (t = 2 h) in both, whereas VLDL1-apoB-100 was maximal at t = 4 h in both, most likely representing delayed VLDL clearance by preferential clearance of chylo and their remnants by competition for the same clearance mechanisms. VLDL2-(Sf20-60)-apoB-48 and VLDL2- apoB-100 decreased in FCHL, in contrast to an increase of apoB-48, and no change of apoB-100 in controls, suggesting impaired conversion of VLDL1-apoB-48 into VLDL2-apoB-48 in FCHL, and partly also of VLDL1-apoB-100. In conclusion, in FCHL clearance of large postprandial Sf >400 apoB-48 and apoB-100 TRL is delayed. ApoB-100 accumulates in the VLDL1 range postprandially in both FCHL and controls, reaching higher levels in FCHL and thereby conferring a higher atherogenic burden in the postprandial situation in FCHL. PMID- 11861670 TI - Differences in reactivity of antibodies to active versus inactive PLTP significantly impacts PLTP measurement. AB - Due to conflicting reports concerning the relationship between phospholipid transfer protein (PLTP) activity and mass in plasma, the protein concentration and activity of PLTP were assessed in fractions isolated by fast protein liquid chromatography from the plasma of healthy normolipidemic individuals. Using both polyclonal and monoclonal antibodies, PLTP was identified by Western blot analysis after both SDS and non-denaturing gradient gel electrophoresis, and quantitated by dot blot. PLTP activity was determined using a labeled vesicle/HDL assay. PLTP mass corresponded substantially with the activity distribution using the polyclonal antibody on dot blot with some inactive PLTP being present. However, the monoclonal antibody preferentially reacted with inactive PLTP, primarily associated with LDL and large HDL, overestimating inactive PLTP. Western blot analysis of non-denaturing gradient gels, using the polyclonal antibody, indicated that active PLTP was associated with numerous discrete HDL subpopulations (7.6-12.0 nm) with the major portion being 9-12 nm. Inactive PLTP was associated with particles of 12 to >17 nm. The monoclonal antibody demonstrated a different pattern of reactivity on gradient gels, showing strong reactivity with the inactive PLTP in particles of 12 to >17 nm, but less reactivity with particles of 7.6-12 nm. The differences in reactivities of antibodies for active versus inactive PLTP can account for some of the discrepancies reported in the literature regarding the relationship between PLTP mass and activity. PMID- 11861671 TI - Microbial biohydrogenation of oleic acid to trans isomers in vitro. AB - Ruminant products are significant sources of dietary trans fatty acids. Trans fatty acids, including various conjugated linoleic acid isomers, have been shown to act as metabolic modifiers of lipid metabolism. Trans fatty acids originate from biohydrogenation of dietary unsaturated fatty acids by gut microbes; however, the exact synthetic pathways are unclear. It was our goal to examine the biohydrogenation pathway for oleic acid, where oleic acid is hydrogenated directly to stearic acid. Our objective in this study was to trace the time course of appearance of 13C in labeled oleic acid to determine if trans monoenes are formed from the 13C-labeled oleic acid or if the 13C appears only in stearic acid as described in reviews of earlier work. Enrichments were calculated from the mass abundance of 13C in major fatty acid fragments and expressed as a percentage of total carbon isotopomers. Significant 13C enrichment was found in stearic acid, oleic acid, trans-6, trans-7, and in all trans C18:1 in positions 9 16. We concluded that the biohydrogenation of oleic acid by mixed ruminal microbes involves the formation of several positional isomers of trans monoenes rather than only direct biohydrogenation to form stearic acid as previously described. PMID- 11861672 TI - The E-box motif in the proximal ABCA1 promoter mediates transcriptional repression of the ABCA1 gene. AB - To identify regulatory elements in the proximal human ATP-binding cassette transporter A1 (hABCA1) gene promoter we transfected RAW cells with plasmids containing mutations in the E-box, AP1, and liver X receptor (LXR) elements as well as the two Sp1 motifs. Point mutations in either Sp1 site or in the AP1 site had only a minor effect whereas mutation of the LXR element decreased promoter activity. In contrast, mutation or deletion of the E-box motif caused a 3-fold increase in transcriptional activity under basal conditions. Gel shift and DNaseI footprint analysis showed binding of a protein or protein complex to this region. Preincubation of nuclear extracts with antibodies established that USF1, USF2, and fos related antigen (Fra) 2 bind to DNA sequences in the human ABCA1 promoter that contains the intact E-box but not the mutant or deleted E-box. Co transfection of USF1 and USF2 enhanced, but Fra2 repressed, ABCA1 promoter activity. Thus, a complex consisting of USF1, USF2, and Fra2 binds the E-box motif 147 bp upstream of the transcriptional start site and facilitates repression of the human ABCA1 promoter. These combined studies identify a novel site in the human ABCA1 promoter involved in the regulation of ABCA1 gene expression. PMID- 11861673 TI - Increased formation of lysophosphatidic acids by lysophospholipase D in serum of hypercholesterolemic rabbits. AB - Lysophosphatidic acid (LPA) is a biologically active phospholipid that has been identified as a vasoactive principle in incubated plasma and serum of mammals. Previously, we found that mammalian plasma and serum contain a lysophospholipase D, which hydrolyzes lysophosphatidylcholines (LPCs) with different fatty acyl groups to the corresponding LPAs during its incubation at 37 degree C. In this study, we examined whether lysophospholipase D activity and levels of LPCs in rabbit serum were modulated by feeding rabbits a high cholesterol diet. Results showed that the serum levels of LPCs increased gradually in animals fed a high cholesterol diet for 12 weeks. We found that the levels of individual LPAs formed on incubation of serum for 24 h increased with an increase in the period of feeding of rabbits a high cholesterol diet. LPA with a linoleate residue was the most abundant LPA, followed in order by 16:0-, 18:1- and 18:0-LPAs. LPA was found to increase attachment of the monocytic cell line THP-1 to vascular endothelial cells pre-stimulated with tumor necrosis factor-alpha. These results indicated that increases in the levels of LPAs generated by lysophospholipase D in the blood of hypercholesterolemic rabbits may be relevant to attachment of monocytes to vascular walls, a key phenomenon observed at an early stage of atherosclerosis. PMID- 11861674 TI - Purification, characterization, and expression of rat intestinal alkaline sphingomyelinase. AB - Intestinal alkaline sphingomyelinase (SMase) has physiological roles in the digestion of sphingomyelin (SM) and clinical implications in colonic carcinogenesis. In the present work, the enzyme from rat has been purified 1,589 fold with 11% recovery by elution of the intestine with bile salt, precipitation of the proteins by acetone, and several types of chromatographies. Its molecular mass was 58 kDa and optimal pH was 9 to 9.5. Under the optimal conditions, the V(max) was 930 micromol/h/mg and K(m) was about 1.25 mM. The enzyme could hydrolyze phosphatidylcholine at pH 7.4 in the presence of Ca2+; the rate was about 8% of that for SM. The activity against SM was dependent on bile salt. Taurine conjugated bile salts were much more effective than glycine conjugated ones, and the most effective bile salts were taurocholate and taurochenodeoxycholate. 3-[(3-Cholamidopropyl) dimethylammonio]-1 propanesulfonate (CHAPS) and Triton X100 (TX100) had no stimulatory effects. Unlike neutral SMase, intestinal alkaline SMase was not Mg2+ dependent, not inhibited by EDTA, and not inhibited by glutathione. The enzyme was stable during incubation with temperatures up to 50 degree C and in pHs from 7 to 10. Trypsin and chymotrypsin had no effects on its activity, and 10 mM dithiothreitol reduced its activity by 25%. A specific antibody against the enzyme was developed, and Western blot showed that the enzyme was expressed in the intestine but not in other organs. In conclusion, we purified a potentially important SMase in the intestine with several properties different from neutral SMase. PMID- 11861675 TI - A new enzyme-linked immunosorbent assay with two monoclonal antibodies to specific epitopes measures human lecithin-cholesterol acyltransferase. AB - We established five monoclonal antibodies that reacted with human LCAT and recognized different epitopes on LCAT. These are mouse anti-human LCAT monoclonal antibodies designated 36487, 36454, 36442, 36405, and 36486, which react with the peptides corresponding to human LCAT amino acid residues R159-E179, M258-S273, S274-S294, D352-S376, and N415-E440, respectively. We also successfully used two of these antibodies to develop an ELISA, which uses a solid phase monoclonal antibody, 36486, that reacts with the C-terminus of LCAT, and a detection monoclonal antibody, 36487, that reacts with an epitope located in the center of the LCAT primary structure. We observed a significant positive correlation between the values of LCAT protein determined with ELISA and LCAT activity determined with liposome substrate (r = 0.871, P < 0.001) or the endogenous self substrate method (r = 0.864, P < 0.001), and we obtained inter- and intra-assay coefficients of variation less than 6.1%, minimum detection limit of 0.1 microg/ml. Highly specific monoclonal antibodies will be useful in the study of the molecular pathology of LCAT. Therefore, this precise and sensitive LCAT assay will help clarify the role of this enzyme in the metabolism of HDLs, and can be used for diagnostic purposes in investigating liver function. We obtained five monoclonal antibodies that recognized different epitopes on LCAT and developed a sandwich-type ELISA. Highly specific monoclonal antibodies provide a sensitive and specific analytical system for measurements of LCAT protein. PMID- 11861676 TI - A rapid single-step centrifugation method for determination of HDL, LDL, and VLDL cholesterol, and TG, and identification of predominant LDL subclass. AB - Determination of the circulating levels of plasma lipoproteins HDL, LDL, and VLDL is critical in the assessment of risk of coronary heart disease. More recently it has become apparent that the LDL subclass pattern is a further important diagnostic parameter. The reference method for separation of plasma lipoproteins is ultracentrifugation. However, current methods often involve prolonged centrifugation steps and use high salt concentrations, which can modify the lipoprotein structure and must be removed before further analysis. To overcome these problems we have now investigated the use of rapid self-generating gradients of iodixanol for separation and analysis of plasma lipoproteins. A protocol is presented in which HDL, LDL, and VLDL, characterized by electron microscopy and agarose gel electophoresis, separate in three bands in a 2.5 h centrifugation step. Recoveries of cholesterol and TG from the gradients were close to 100%. The distribution profiles of cholesterol and TG in the gradient were used to calculate the concentrations of individual lipoprotein classes. The values correlated with those obtained using commercial kits for HDL and LDL cholesterol. The position of the LDL peak in the gradient and its shape varied between plasma samples and was indicative of the density of the predominant LDL class. The novel protocol offers a rapid, reproducible and accurate single-step centrifugation method for the determination of HDL, LDL, and VLDL cholesterol, and TG, and identification of LDL subclass pattern. PMID- 11861677 TI - Oxazolinone derivative of leucine for GC-MS: a sensitive and robust method for stable isotope kinetic studies of lipoproteins. AB - Stable isotope labeled amino acids are commonly used as endogenous tracers to study the metabolism of lipoproteins. The determination of isotopic enrichment of particular amino acids in apolipoproteins is carried out by gas chromatography mass spectrometry (GC-MS). This report describes a robust and sensitive derivative for analysis of d3-leucine by GC-MS and its utility in studying the metabolism of human lipoproteins. The trifluoromethyloxazolinone (oxazolinone) derivative of leucine was formed in a rapid single step procedure using a mixture of trifluoroacetic anhydride (TFAA) and trifluoroacetic acid (TFA). Analysis of the oxazolinone by negative ion chemical ionization GC-MS gave excellent sensitivity and precision, which enabled accurate determination of low levels of isotopic enrichment from small amounts of protein. For example, enrichments between 0.05% and 100% in 100 pg leucine can be measured with a coefficient of variation of <3%. To demonstrate the utility of this procedure, we measured d3 leucine enrichment in apolipoprotein B (apoB) isolated from VLDL and LDL as well as apoA-I isolated from HDL by gel electrophoresis and western blotting. The derivatization procedure gave excellent enrichment data from a single intravenous bolus dose of 5 mg/kg, from which the fractional catabolic rate and production rate of the lipoproteins were calculated. In conclusion, the oxazolinone derivative provides a robust and simple procedure for the sensitive analysis of isotopic enrichment for metabolic studies of human lipoproteins. PMID- 11861678 TI - Simplified methodology to determine breast milk retinol concentrations. AB - The impact of a nutritional intervention trial on vitamin A status can be evaluated by measuring the total vitamin A concentration in breast milk both before and after the intervention. Because breast milk contains a spectrum of retinyl esters as well as retinol and high lipid content, determination of total breast milk retinol routinely requires saponification with alcoholic potassium hydroxide. Retinol is then extracted with an organic solvent, usually hexanes, before HPLC analysis. Retinyl acetate, although commonly used as an internal standard, is not ideal because it can only be added after saponification and extraction and consequently, will only account for part of the total losses. A method has now been developed that uses 3,4-didehydroretinyl acetate (DRA) as an internal standard. DRA is an excellent choice as an internal standard for the following reasons: 1) DRA can be added to the breast milk before saponification and can be carried through the analysis as dehydroretinol (DR), 2) the percent recovery can be easily determined, and 3) DR is easily separated from retinol during HPLC analysis. The procedure, as described, typically gives a mean extraction efficiency of 80-90%. Moreover, the average coefficient of variation is <5% on the same sample run several times in the same day. PMID- 11861679 TI - Relevance of new psychotropic drugs for the neurologist. PMID- 11861680 TI - Variant Creutzfeldt-Jakob disease. PMID- 11861681 TI - Dystonia in multiple system atrophy. PMID- 11861683 TI - Raeder's syndrome [corrected]: paratrigeminal paralysis of the oculopupillary sympathetic system. AB - Raeder described five patients with mixed features of trigeminal nerve pathology and oculosympathetic impairment, with or without other cranial nerve lesions. This constellation of clinical features drew the original author's attention to the paratrigeminal region as a likely site for the causative lesion in this syndrome. An analysis of the anatomy of the oculosympathetic innervation supports the view that a restricted lesion in the middle cranial fossa might cause the syndrome of trigeminal nerve involvement, neuralgic pain or sensory change, with ptosis or miosis, or both, but no anhidrosis. Such a paratrigeminal oculosympathetic syndrome (POSS) usefully reminds clinicians to pursue vigorously possible lesions of the middle cranial fossa with careful, and possibly repeated, imaging studies. Attaching the eponym Raeder's syndrome or Raeder's paratrigeminal neuralgia to this syndrome adds nothing valuable to the anatomical description (POSS), which might be preferred for clarity. PMID- 11861682 TI - Genetic effects on human cognition: lessons from the study of mental retardation syndromes. AB - The molecular basis of human cognition is still poorly understood, but recent advances in finding genetic mutations that result in cognitive impairment may provide insights into the neurobiology of cognitive function. Here we review the progress that has been made so far and assess what has been learnt from this work on the relation between genes and cognitive processes. We review evidence that the pathway from genetic lesion to cognitive impairment can be dissected, that some genetic effects on cognition are relatively direct and we argue that the study of mental retardation syndromes is giving us new clues about the biological bases of cognition. PMID- 11861684 TI - Dystonia in multiple system atrophy. AB - OBJECTIVE: To delineate the frequency and nature of dystonia in multiple system atrophy (MSA). METHODS: A cohort of 24 patients with clinically probable MSA over the past 10 years were prospectively followed up. Motor features were either dominated by parkinsonism (MSA-P subtype, n=18) or cerebellar ataxia (MSA-C, n=6). Classification of dystonic features and their changes with time was based on clinical observation during 6-12 monthly follow up visits. Parkinsonian features and complications of drug therapy were assessed. Most patients (22/24) died during the observation period. Neuropathological examination was confirmatory in all of the five necropsied patients. RESULTS: At first neurological visit dystonia was present in 11 (46%) patients all of whom had been levodopa naive at this time point. Six patients (25%) exhibited cervical dystonia (antecollis) (MSA-P n=4, MSA-C n=2), five patients (21%) showed unilateral limb dystonia (MSA-P n=4; MSA-C n=1). A definite initial response to levodopa treatment was seen in 15/18 patients with MSA-P, but in none of the six patients with MSA-C. A subgroup of 12 patients with MSA-P developed levodopa induced dyskinesias 2.3 years (range 0.5-4) after initiation of levodopa therapy. Most patients had peak dose craniocervical dystonia; however, some patients experienced limb or generalised dystonia. Isolated peak dose limb chorea occurred in only one patient. CONCLUSION: The prospective clinical study suggests that dystonia is common in untreated MSA-P. This finding may reflect younger age at disease onset and putaminal pathology in MSA-P. Levodopa induced dyskinesias were almost exclusively dystonic affecting predominantly craniocervical musculature. Future studies are required to elucidate the underlying pathophysiology of dystonia in MSA. PMID- 11861685 TI - Possible underascertainment of variant Creutzfeldt-Jakob disease: a systematic study. AB - OBJECTIVES: To predict the size of the vCJD epidemic it is important to know whether the description of cases of vCJD in 1996 represent the first cases of a new disease entity or whether detection was due to increased surveillance of CJD in humans. Detection of earlier cases would suggest a shorter incubation period and might lead to predictions of epidemic size being revised. METHODS: All certified deaths (excluding external injury and poisoning) in residents of Wales aged 15-45, between 1985 and 1995, were reviewed to detect vCJD deaths that might have been overlooked. 12 091 deaths were reviewed. "Non-specific fatal disorders compatible with vCJD" were defined. Deaths recorded to diseases other than those defined were rejected from further analysis (8769). Remaining cases (3322) were subdivided. Group A comprised deaths recorded to suicide, transport accidents, and those that could not be ascertained (ICD9 rubrics E950-959, E800-848, and 7999), a total of 2698 cases. Group B comprised deaths due to neurological disease, psychiatric disease, or substance abuse (624). RESULTS: For group A, remaining brain material was identified (n=218, 8.1%) and examined by routine histology and immunocytochemistry for prion protein. No cases of vCJD were detected. For group B, review of remaining clinical information was undertaken. Of 624 cases, information was available on 447 (72%). Brain tissue was examined by routine histology and immunocytochemistry in 47 (7.5%) cases. Sufficient clinical and pathological information was available to exclude all these as potential cases of vCJD. CONCLUSION: Variant CJD is a new disease entity and not simply the result of better case ascertainment. PMID- 11861686 TI - Cholinesterase inhibitor treatment alters the natural history of Alzheimer's disease. AB - OBJECTIVE: To describe the effect of cholinesterase inhibitors (CEIs) on the natural course of Alzheimer's disease (AD). METHODS: The short and long term effects of CEIs were evaluated in 135 patients with probable Alzheimer's disease relative to 135 patients who were never exposed to CEIs matched by age, education, duration of the symptoms, and cognitive status. We measured 1 year change in cognitive and functional performance, and the likelihood of arriving at each of four end points: (1) mini mental state examination (MMSE) of 9 or lower, (2) Blessed dementia rating scale for activities of daily living of 12 or higher, (3) nursing home admission, and (4) death, over an average 3 years of observation (36.7 (SD 21.5) months). RESULTS: Patients on CEIs were better cognitively and functionally after 1 year compared with those patients who never used CEIs. A proportional hazard analysis with CEI use as a time dependent covariate showed that the use of CEIs decreased the risk of nursing home admission. There was no association, however, between use of CEIs and time to cognitive and functional end points, or to death. CONCLUSIONS: This observational study showed that there was an initial cognitive and functional benefit from the use of CEIs in Alzheimer's disease, which waned as the disease progressed. However, the results suggest that there is a long term beneficial effect of the use of CEIs, as indicated by the delay in admission to nursing homes. PMID- 11861687 TI - Adaptation of handwriting size under distorted visual feedback in patients with Parkinson's disease and elderly and young controls. AB - OBJECTIVE: The ability to use visual feedback to control handwriting size was compared in patients with Parkinson's disease (PD), elderly people, and young adults to better understand factors playing a part in parkinsonian micrographia. METHODS: The participants wrote sequences of eight cursive l loops with visual target sizes of 0.5 and 2 cm on a flat panel display digitiser which both recorded and displayed the pen movements. In the pre-exposure and postexposure conditions, the display digitiser showed the actual pen trace in real time and real size. In the distortion exposure conditions, the gain of the vertical dimension of the visual feedback was either reduced to 70% or enlarged to 140%. RESULTS: The young controls showed a gradual visuomotor adaptation that compensated for the visual feedback distortions during the exposure conditions. They also showed significant after effects during the postexposure conditions. The elderly controls marginally corrected for the size distortions and showed small after effects. The patients with PD, however, showed no trial by trial adaptations or after effects but instead, a progressive amplification of the distortion effect in each individual trial. CONCLUSION: The young controls used visual feedback to update their visuomotor map. The elderly controls seemed to make little use of visual feedback. The patients with Parkinson's disease rely on the visual feedback of previous or of ongoing strokes to programme subsequent strokes. This recursive feedback may play a part in the progressive reductions in handwriting size found in parkinsonian micrographia. PMID- 11861688 TI - Neurophysiological modulation of the subthalamic nucleus by pallidal stimulation in Parkinson's disease. AB - OBJECTIVES: Current models of basal ganglia dysfunction in Parkinson's disease suggest a pivotal role of subthalamic nucleus (STN) hyperactivity. There is a direct excitatory output to the globus pallidus internus (GPi), which in turn hyperinhibits the motor thalamus and leads to a lack of cortical facilitation. The model, however, does not address the reciprocal influence of GPi on STN activity. METHODS: Measurement of immediate changes in STN single cell activity after GPi deep brain stimulation (DBS). RESULTS: An opposite effect of GPi DBS in the dorsal versus ventral STN was found. There was an almost exclusive reduction of firing rate in the dorsal region of the STN, whereas the cells in the ventral region exhibited facilitation similar to the recordings from the substantia nigra pars reticulata. CONCLUSION: Although these findings require confirmation, they suggest that the current theories of GPi DBS action, which do not include a GPi STN modulation, are most likely incomplete. PMID- 11861689 TI - Evidence of thalamic disinhibition in patients with hemichorea: semiquantitative analysis using SPECT. AB - OBJECTIVES: Hemichorea sometimes occurs after lesions that selectively involve the caudate nucleus, putamen, and globus pallidus. Some reports have hypothesised that the loss of subthalamic nucleus control on the internal segment of the globus pallidus, followed by the disinhibition of the thalamus may contribute to chorea. However, the pathophysiology is poorly understood. Therefore, clinicoradiological localisation was evaluated and a comparison of the haemodynamic status of the basal ganglia and thalamus was made. METHODS: Six patients presenting with acute onset of hemichorea were assessed. Neuroimaging studies, including MRI and SPECT examinations in addition to detailed biochemical tests, were performed. A semiquantitative analysis was performed by comparing the ratio of blood flow between patients and normal controls. In addition, the ratio of perfusion asymmetry was calculated as the ratio between each area contralateral to the chorea and that homolateral to the chorea. The comparison was made with a two sample t test. RESULTS: The causes of hemichorea found consisted of four cases of acute stroke, one non-ketotic hyperglycaemia, and one systemic lupus erythematosus. Brain MRI indicated lesion sites in the contralateral putamen, globus pallidus, caudate nucleus, and subthalamic nucleus. A significant decrease in the ratio of blood flow in the basal ganglia contralateral to the chorea and a significant increase in the thalamus was found when comparing the perfusion asymmetries, which were calculated as the ratio of cerebral blood flow (CBF) for each region to that in the homolateral occipital area (p<0.05). CONCLUSION: An alteration in CBF in both the contralateral thalamus and basal ganglia reflect the loss of pallidal inhibitory input from the pallidum to the thalamus. This change in CBF may be one of epiphenomena, which implicates an occurrence of hemichorea in humans. PMID- 11861690 TI - Aneurysmal subarachnoid haemorrhage: outcomes of early rehabilitation after surgical repair of ruptured intracranial aneurysms. AB - OBJECTIVES: The aim was to analyse functional and cognitive outcomes in patients receiving early rehabilitation treatment after surgery for aneurysmal subarachnoid haemorrhage (SAH). METHODS: The assessment protocol included all relevant clinical data, the Hunt-Hess scale, the functional independence measure (FIM), and the mini mental state examination (MMSE). RESULTS: Of 59 patients included in the study, 52.5% (31) were men and 47.5% (28) were women. The patients' average age was 52 years, and 57.6% were employed at the time of the aneurysm rupture. The mean duration of hospital stay was 25 days; 67.8% (40) of the patients were discharged home. At discharge, 72.7% of the patients were without any motor impairment, but 59.6% showed cognitive impairment. By the time of discharge, 43.4% (23) of the patients had attained independence in activites of daily living, 18.9% (10) needed intermittent supervision, and 37.7% (20) required constant supervision in the performance of these activities. CONCLUSIONS: The severity of cognitive impairment has predictive value for the functional status and the level of supervision required at discharge. PMID- 11861691 TI - Cessation of embolic signals after antithrombotic prevention is related to reduced risk of recurrent arterioembolic transient ischaemic attack and stroke. AB - OBJECTIVES: To evaluate the reduction of embolic signals after the initiation of an antithrombotic secondary prevention in patients with recent arterioembolic stroke and to determine the predictive value of decreased microembolism on the risk of early stroke recurrence. METHODS: Eighty six consecutive patients (55 men, 31 women; mean age 60.6 years) with a non-disabling arterioembolic ischaemic event in the anterior circulation within the last 30 days and a medium grade or high grade stenosis (> or =50%) of the ipsilateral carotid or middle cerebral artery underwent 1 hour transcranial Doppler monitoring as part of the admission examinations. Antithrombotic secondary prevention was started after completion of admission. Patients in whom embolic signals were detected underwent a second monitoring within 4 days (mean time 1.8 days). All patients were followed up prospectively to evaluate the relation between presence and persistence of embolic signals and the risk of recurrent transient ischaemic attack (TIA) and stroke within the next 6 weeks. RESULTS: In 44 patients, embolic signals were detected at admission, a mean 5.4 days (range 0 to 21 days) after the initial event. Twenty five were positive for embolic signals also at the second monitoring, in 19 signals had ceased. Forty two patients without embolic signals at admission served as controls. During follow up, six ischaemic events (two stroke, three TIA, one amaurosis fugax) occurred in 25 patients with persisting embolic signals but none in 19 patients in whom signals had ceased by the second monitoring. One patient in the control group had a TIA. The incidence of a recurrent event was 0.45 per 30 patient-days if embolic signals persisted compared with 0.015 if signals could not be detected or had ceased. Persistence of embolic signals was an independent predictor of a recurrent TIA or stroke (adjusted odds ratio 37.0; 95% confidence interval (95% CI) 3.5 to 333; p<0.003). Cessation and decrease of embolic signals was associated with the administration of antiplatelet agents but not with anticoagulation with intravenous heparin (p<0.001). CONCLUSIONS: Rapid cessation of embolic signals detected in patients with recently symptomatic arterial stenosis decreases increased risk of an early ischaemic recurrence. Effect of antithrombotic agents on embolic signals might serve as a marker for their efficacy on preventing stroke recurrence. PMID- 11861692 TI - Angiotensin converting enzyme insertion/deletion genotype is associated with leukoaraiosis in lacunar syndromes. AB - OBJECTIVES: Pathological and clinical data suggest that patients presenting with ischaemic lacunar syndromes may be a heterogenous group. Those with isolated lacunar infarction are thought to have localised atherosclerosis whereas in those with coexisting leukoaraiois a distinct diffuse small vessel vasculopathy may be the predominant underlying pathology. The ACE insertion/deletion (I/D) polymorphism is an important candidate gene in ischaemic cerebrovascular disease but, where lacunar stroke specifically has been examined, there have been discrepant reports concerning a possible association. It was hypothesised that the influence of the ACE gene may be different among the two subgroups of ischaemic lacunar stroke reflecting the heterogeneity of the small vessel disease phenotype. METHODS: Eighty four consecutive patients presenting with classic lacunar syndromes were studied. All had acute cranial CT to exclude primary intracerebral haemorrhage and these were subsequently assessed for the presence and extent of leukoaraiosis. All patients were genotyped for the ACE insertion/deletion polymorphism. RESULTS: There was a significant difference in the distribution of ACE genotype with the DD genotype occurring more often in patients with leukoaraiosis and the II and ID genotypes occurring more often among those in whom this was absent (chi(2)=9.06, p=0.01). In a logistic regression model the ACE DD genotype remained as an independent predictor for the presence of leukoaraiosis (p=0.02) in patients presenting with classic lacunar syndromes. CONCLUSION: This study supports the hypothesis that there may be different types of small vessel disease in patients with classic lacunar syndromes and that the influence of the ACE DD genotype may be relevant in mediating the diffuse form of vessel injury. PMID- 11861693 TI - Outcome of patients with cryptogenic stroke and patent foramen ovale. AB - OBJECTIVES: The aim was to estimate the recurrence rate and to define subgroups at increased risk for recurrent cerebral ischaemia in patients with patent foramen ovale (PFO) and so called cryptogenic stroke due to paradoxical embolism. METHODS: Patent foramen ovale was diagnosed in 318 patients with otherwise unexplained ischaemic stroke or transient ischaemic attack (TIA). One hundred and fifty nine were treated medically (oral anticoagulation 79, platelet inhibitors 80) and represent the study population. The remaining 159 patients underwent endovascular or surgical closure of the PFO and are not part of this study. RESULTS: Mean age was 50.7 (SD 13.5) years. The event leading to the diagnosis of PFO was a TIA in 38 patients (23.9%), an ischaemic stroke in 119 (74.8%), and an amaurosis fugax in two patients (1.3%). Forty four patients (27.7%) had experienced multiple cerebrovascular ischaemic events before the diagnosis of the PFO. During mean follow up of 29 (SD 23) months 21 patients (13.4%) had a recurrent cerebrovascular event (seven strokes and 14 TIAs). The average annual rate of recurrent strokes was 1.8% and that of recurrent strokes or TIAs was 5.5%. When patients with PFO with multiple cerebrovascular events before the diagnosis of the PFO were analyzed separately, the average annual rates of recurrent cerebral ischaemia were 3.6% for recurrent strokes and 9.9% for recurrent strokes or TIAs. These rates were significantly higher than in patients with first ever stroke or TIA (p=0.02). CONCLUSIONS: The study confirms a risk of stroke recurrence that is similar to the rates of previously published series of patients with PFO and cryptogenic strokes. Patients with more than one previous event were at increased risk of recurrent cerebral ischaemia. PMID- 11861694 TI - Surgical removal of brain stem cavernous malformations: surgical indications, technical considerations, and results. AB - OBJECTIVES: This study was undertaken to review the indications for surgical treatment of brain stem cavernomas and to develop strategies to minimise the complications of surgery. PATIENTS AND RESULTS: Twelve patients underwent surgical resection of a brain stem cavernoma due to symptoms caused by one or more haemorrhages. Age ranged from 18 to 47 years (mean 29.2 years). Long term follow up (mean 3.7 years) included a complete neurological examination and annual MRI studies. The annual haemorrhage rate was 6.8 %/patient/year and a rate of 1.9 rehaemorrhages/patient/year was found. Surgery was performed under microsurgical conditions with endoscopic assistance, use of neuronavigation, and neurophysiological monitoring. Navigation proved to be reliable when applied in an early stage of operative procedure with minimal brain retraction. Endoscopy was a useful tool in some cases to confirm complete resection of the lesion and to ascertain haemostasis. Ten patients had a new neurological deficit in the early postoperative period, nine of these were transient. At the last follow up the neurological state was improved in five patients, unchanged in six, and worse in one compared with the preoperative conditions. The preoperative average Rankin score was 2.2 points and had improved at the last follow up by 0.6 points to 1.6 points. CONCLUSIONS: Symptomatic brain stem cavernomas should be considered for surgical treatment after the first bleeding. Careful selection of the optimal operative approach and a meticulous microsurgical technique are mandatory. The additional use of modern tools such as neuronavigation, endoscopic assistance, and monitoring can contribute to the safety of the procedure. PMID- 11861697 TI - Changes in gait and fatigue from morning to afternoon in people with multiple sclerosis. AB - OBJECTIVES: The aim was to measure changes in walking patterns and self rated fatigue in people with multiple sclerosis (MS) compared with age matched control subjects, from the morning to the afternoon within a single day. METHODS: Fourteen patients with MS and the same number of matched control subjects performed four 10 m gait trials at their preferred walking speed at 10 00 am and then again at 3 00 pm on the same day. Gait speed, stride length, cadence, and the percentage of the gait cycle spent in double limb support were measured using a foot switch stride analyzer. Patients with MS also self rated their fatigue levels in the morning and afternoon using an 11 point scale. RESULTS: Compared with control subjects, patients walked very slowly, with reduced stride length and around twice as much variability in gait performance. Although self rated fatigue significantly increased from the morning to the afternoon, walking patterns remained consistent in both groups over the course of the day. CONCLUSIONS: These findings imply that mechanisms controlling locomotion are separate from those regulating perceived fatigue. Objective measures of performance, rather than self report, should be used to monitor change in patients with multiple sclerosis. PMID- 11861696 TI - Organisation of the sympathetic skin response in spinal cord injury. AB - OBJECTIVES: The sympathetic skin response (SSR) is a technique to assess the sympathetic cholinergic pathways, and it can be used to study the central sympathetic pathways in spinal cord injury (SCI). This study investigated the capacity of the isolated spinal cord to generate an SSR, and determined the relation between SSR, levels of spinal cord lesion, and supraspinal connections. METHODS: Palmar and plantar SSR to peripheral nerve electrical stimulation (median or supraorbital nerve above the lesion, and peroneal nerve below the lesion) were recorded in 29 patients with SCI at various neurological levels and in 10 healthy control subjects. RESULTS: In complete SCI at any neurological level, SSR was absent below the lesion. Palmar SSR to median nerve stimuli was absent in complete SCI with level of lesion above T6. Plantar SSR was absent in all patients with complete SCI at the cervical and thoracic level. In incomplete SCI, the occurrence of SSR was dependent on the preservation of supraspinal connections. For all stimulated nerves, there was no difference between recording from ipsilateral and contralateral limbs. CONCLUSIONS: No evidence was found to support the hypothesis that the spinal cord isolated from the brain stem could generate an SSR. The results indicate that supraspinal connections are necessary for the SSR, together with integrity of central sympathetic pathways of the upper thoracic segments for palmar SSR, and possibly all thoracic segments for plantar SSR. PMID- 11861698 TI - Positional down beating nystagmus in 50 patients: cerebellar disorders and possible anterior semicircular canalithiasis. AB - OBJECTIVES: To clarify the clinical significance of positional down beat nystagmus (pDBN). METHODS: A discussion of the neuro-otological findings in 50 consecutive patients with pDBN. RESULTS: In 38 patients there was evidence of CNS disease (central group) but in 12 there was not (idiopathic group). In the CNS group, presenting symptoms were gait, speech, and autonomic dysfunction whereas in the idiopathic group patients mostly reported positional vertigo. The main neurological and oculomotor signs in the CNS group were explained by cerebellar dysfunction, including 13 patients with multiple system atrophy. In patients with multiple system atrophy with a prominent extrapyramidal component, the presence of pDBN was helpful in the differential diagnosis of atypical parkinsonism. No patient with pDBN had the Arnold-Chiari malformation, a common cause of constant down beat nystagmus (DBN). In the idiopathic group, the pDBN had characteristics which suggested a peripheral labyrinthine disorder: vertigo, adaptation, and habituation. In six patients an additional torsional component was found (concurrently with the pDBN in three). Features unusual for peripheral disorder were: bilateral positive Dix-Hallpike manoeuvre in nine of 12 patients and selective provocation by the straight head-hanging manoeuvre in two. CONCLUSION: It is argued that some patients with idiopathic pDBN have benign paroxysmal positional vertigo (BPPV) with lithiasis of the anterior canal. The torsional component may be weak, because of the predominantly sagittal orientation of the anterior canal, and may not be readily seen clinically. Nystagmus provocation by bilateral Dix-Hallpike and straight head-hanging may be explained by the vertical upwards orientation of the ampullary segment of the anterior canal in the normal upright head position. Such orientation makes right-left specificity with the Dix Hallpike manoeuvre less important than for posterior canal BPPV. This orientation requires a further downwards movement of the head, often achieved with the straight head-hanging position, to provoke migration of the canaliths. The straight head-hanging manoeuvre should be carried out in all patients with a history of positional vertigo and a negative Dix-Hallpike manoeuvre. PMID- 11861699 TI - Effect of brain tumour laterality on patients' perceived quality of life. AB - OBJECTIVES: There is little reliable quantitative information on preoperative quality of life of patients with brain tumours. The aim of this study was to clarify the effect of the volume, location, and histological grade of brain tumours on the preoperative quality of life of patients. METHODS: The study population consisted of 101 successive patients with brain tumour at Oulu Clinic for Neurosurgery studied with CT or MRI for preoperative determination of tumour location and size. The Nottingham health profile (NHP) and Sintonen's 15D scale were used at that time to measure quality of life. RESULTS: Tumour size did not correlate linearly with impairment of quality of life. Large tumours (>25 ml) were associated with poorer quality of life than small tumours (< or =25 ml). The patients with a tumour located on the right side or in the anterior region reported a poorer quality of life than those with a tumour on the left side or posteriorly. Quality of life assessments made by doctors using the Karnofsky performance scale showed no differences between the two hemispheres. Patients with the most malignant gliomas (grades III-IV) displayed the poorest quality of life. CONCLUSIONS: Large tumours apparently damage several parts of the brain and/or raise intracranial pressure to a level that exceeds the brain's compensatory capacity. Contrary to earlier understanding, tumours in the right hemisphere seemed to be related to poorer quality of life. This effect was especially clear in the patients' subjective evaluation of their quality of life. As the location of the brain tumour thus affects perceived quality of life, any measurements of the quality of life of patients with brain tumours should take into account the location and laterality of the tumour. PMID- 11861701 TI - Notch3 gene polymorphism and ischaemic cerebrovascular disease. AB - Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is a type of hereditary stroke and dementia. More than 90% of patients with CADASIL have mutations in the Notch3 gene. All mutations either create or destroy a cysteine residue in the epidermal growth factor-like repeats. In addition, five polymorphisms, which lead to amino acid substitutions, have been identified within the Notch3 coding sequence. However, whether these polymorphisms affect Notch signalling or are involved in cerebrovascular diseases is unknown. In the present study, we investigated a possible association between a T6746C polymorphism in the Notch3 coding region and the occurrence of symptomatic ischaemic cerebrovascular disease (CVD) was investigated. Two hundred and thirty five patients with CVD, as confirmed by brain CT or MRI, and 315 age and sex matched control subjects were analyzed for genotype frequencies of the T6746C polymorphism in Notch3. The genotype distributions were: patients with CVD, C/C 14.0%, C/T 45.5%, and T/T 40.4%; controls, C/C, 14.3%; C/T, 47.9%; T/T, 37.8%. The Japanese population has a higher C allele frequency of the T6746C polymorphism than European populations. There was no significant difference between the T6746C polymorphism in patients with CVD and controls (chi(2)=0.414, p=0.813). This was confirmed by the results of multiple logistic regression analysis including established risk factors (chi(2) =4.65, p=0.311). In conclusion, the results indicate that T6746C polymorphism in the intracellular domain of the Notch3 gene is not associated with an increased risk for CVD. PMID- 11861700 TI - Use of neuronavigation and electrophysiology in surgery of subcortically located lesions in the sensorimotor strip. AB - OBJECTIVES: Subcortical lesions in the sensorimotor strip are often considered to be inoperable. The purpose of this study was to evaluate the usefulness of a combined approach for surgery in this region, aided by a robotic neuronavigation system under electrophysiological control. METHODS: In a prospective study on 10 patients, space occupying lesions in the sensorimotor central area were removed using the Surgiscope robotic navigation system and the Nicolet Viking IV electrophysiological system. RESULTS: Precise tumour localisation with the neuronavigation system and the information on the patient's cortical motor distribution obtained by bipolar cortical stimulation led to postoperative improvement in motor function in all but one patient. Seven of the patients had focal, defined pathology (four metastases; two cavernoma; one aspergilloma). CONCLUSION: Due to the implementation of two recent technologies, surgery of lesions in the subcortical sensorimotor region can be performed with greater confidence. PMID- 11861702 TI - Changes in third ventricular size with neuroendoscopic third ventriculostomy: a blinded study. AB - The objective was to study the relation between changes in cerebral ventricular size and clinical outcome after neuroendoscopic third ventriculostomy (NTV) in both primary (no previous surgery) and secondary (previous CSF diversion for hydrocephalus) NTV. Changes in ventricular size were related to the need for further surgery for CSF diversion. A blinded retrospective study of the pre-NTV and post-NTV sizes of the ventricles in an unselected series of patients undergoing this procedure was done. A decrease in third ventricular size was seen in significantly more patients in the primary group than in the secondary group. Median change in third ventricle width for those who did not require further CSF diversion was significantly greater than those with no clinical benefit (p=0.01). Positive predictive power for successful outcome was highest for measures of the third ventricle; 73% and 68% for third ventricle width and height respectively, 88% for anterior to posterior commissure distance. In conclusion, third ventricular size reduces after NTV. The use of such a change as an arbiter of success in this procedure is questionable as clinically successful cases can have no change in ventricular size. It is considered that clinical outcome is the most important guide to success or failure as reduction in ventricular size is by no means guaranteed. Radiological outcomes alone may be misleading and reliance on them should be avoided. PMID- 11861703 TI - Sequence analysis of tau in familial and sporadic progressive supranuclear palsy. AB - Progressive supranuclear palsy (PSP) is a tau deposition neurodegenerative disorder which usually occurs in sporadic form and is associated with a common variant of the tau gene. Rare familial forms of PSP have been described. Recently familial frontotemporal dementia linked to chromosome 17 (FTDP-17) has been shown to be due to mutations in tau and there may be a clinical and pathological overlap between PSP and FTDP-17. In this study we have analysed the tau sequence in two small families with PSP, and a number of clinically typical and atypical sporadic cases with pathological confirmation of the diagnosis. The tau mutations described in FTDP-17 were not found in the most clinically diagnosed patients with PSP. This suggests that usually FTDP-17 and PSP, including the rare familial form of PSP, are likely to be separate conditions and that usually PSP and typical PSP-like syndromes are not due to mutations in tau. PMID- 11861705 TI - Clipping of cerebral aneurysm under hypothermic cardiac arrest and simultaneous coronary artery bypass grafting: case report. AB - This report describes a case where joint neurological and cardiac surgery teams cooperated to perform simultaneous procedures of clipping a complex internal carotid artery under hypothermic cardiac arrest and coronary artery bypass grafting. A 69 year old man was evaluated for complaints of double vision, pain behind his right eye, and progressively worsening headaches. Examination showed bilateral upgoing toes and difficulty performing a tandem gait. The patient had a history of myocardial infarction. Brain MRI showed a 1.6 cm diameter, partially thrombosed aneurysm of the right internal carotid artery and posterior communicating artery. Cardiac catheterisation showed critical coronary artery disease of the distal segment of the right coronary artery with 90% stenosis of the midcircumflex artery and an ejection fraction of 40%. After initial exposure of the aneurysm, the cardiac team instituted hypothermic cardiac arrest (21 degree C). The aneurysmal sac was collapsed and dissected from the surrounding perforators. An encircling fenestrated clip was applied and a small part of the neck of the aneurysm was further clipped with straight clips. The cardiac surgery team performed the coronary artery bypass grafting procedure. The patient recovered fully, returned to his normal activities, and is functioning independently. PMID- 11861704 TI - Peripheral facial palsy and dysfunction of the oropharynx. AB - The facial nerve contributes to the oropharyngeal phase of deglutition via the buccinator, perioral, digastricus posterior, and stylohyoid muscles. The gustatory and salivatory functions of the facial nerve are also known to contribute to swallowing. The relation between peripheral facial nerve palsy (PFP) and swallowing dysfunction has never been studied systematically. Forty four patients with unilateral Bell's palsy (acute or chronic stages) and 20 normal control subjects were investigated. In 66% of patients with PFP, oropharyngeal swallowing was disturbed as demonstrated electrophysiologically by the patients' dysphagia limit at or below 20 ml of water. In patients with PFP investigated within the first 2 weeks of the palsy, the dysphagia limit normalised during the period of recovery. Normalisation of the dysphagia limit is highly correlated with the recovery of PFP. Thus, subclinical deglutition is very frequent in patients with PFP. The severity of abnormal deglutition increased with the severity of the PFP, especially with the involvement of the perioral and buccinator muscles. PMID- 11861706 TI - A new defect of peroxisomal function involving pristanic acid: a case report. AB - AN adult onset novel disorder of peroxisomal function is described, characterised by retinitis pigmentosa resulting in progressive visual failure, learning difficulties, a peripheral neuropathy, and hypogonadism. The defect results in accumulation of pristanic acid, and the bile acid intermediates, dihydroxycholestanoic and trihydroxycholestanoic acid, and is due to a deficiency of alpha-methylacyl-CoA racemase, making this the first fully characterised description of this defect. Screening of patients with retinitis pigmentosa should be extended to include pristanic acid and/or bile acid intermediate concentrations, as dietary measures offer a potential treatment for the disorder. PMID- 11861707 TI - Rumination and executive function in depression: an experimental study. AB - BACKGROUND: Major depression is associated with cognitive deficits, particularly those requiring central executive functioning. Depressed patients also tend to focus on and think about their symptoms and problems ("ruminate") more than non depressed controls. Although an association has been found between rumination and impaired performance on a central executive processing task, the causal relation between impaired executive functioning and rumination has not been determined. This study sought to directly manipulate rumination and assess the impact on executive functioning in depression as measured by random number generation. METHODS: Depressed patients (n=14) and non-depressed controls (n=14) were compared on a random number generation task, performed after both a rumination induction and after a distraction induction, with order of inductions counter balanced within each group. RESULTS: Compared with the distraction induction, the rumination induction produced a significant increase in both ruminations and the tendency towards stereotyped counting responses (thought to reflect a failure of inhibitory executive control) in depressed patients but not in controls. However, after distraction, no difference in random number generation or rumination was found between the two groups. CONCLUSIONS: The aspects of executive function involved in random number generation are not fundamentally impaired in depressed patients. In depressed patients, the rumination induction seems to trigger the continued generation of ruminative stimulus independent thoughts, which interferes with concurrent executive processing. PMID- 11861708 TI - The Gerstmann syndrome in Alzheimer's disease. AB - BACKGROUND: It remains unclear from lesion studies whether the four signs of the Gerstmann syndrome (finger agnosia, acalculia, agraphia, and right-left confusion) cluster because the neuronal nets that mediate these activities have anatomical proximity, or because these four functions share a common network. If there is a common network, with degeneration, as may occur in Alzheimer's disease, each of the signs associated with Gerstmann's syndrome should correlate with the other three signs more closely than they correlate with other cognitive deficits. METHODS: Thirty eight patients with probable Alzheimer's disease were included in a retrospective analysis of neuropsychological functions. RESULTS: The four Gerstmann's syndrome signs did not cluster together. Finger naming and calculations were not significantly correlated. Right-left knowledge and calculations also did not correlate. CONCLUSIONS: The four cognitive functions impaired in Gerstmann's syndrome do not share a common neuronal network, and their co-occurrence with dominant parietal lobe injuries may be related to the anatomical proximity of the different networks mediating these functions. PMID- 11861709 TI - Volumetric MRI study of the caudate nucleus in patients with dementia with Lewy bodies, Alzheimer's disease, and vascular dementia. AB - OBJECTIVES: To determine whether parkinsonian symptoms in dementia with Lewy bodies (DLB) are associated with greater atrophy of the caudate nucleus in comparison with patients with Alzheimer's disease (AD) and vascular dementia (VaD). METHODS: T1weighted MR scans were acquired in elderly patients with DLB, AD, VaD, and healthy controls. Normalised volumetric measurements of the caudate nucleus were obtained and parkinsonian symptoms rated using Hoehn and Yahr staging. RESULTS: There were no significant differences in the volume of the caudate nucleus between patients with dementia. However, the left caudate volume was significantly reduced in AD and DLB compared with controls. Parkinsonian symptoms did not correlate with caudate nucleus volume. CONCLUSIONS: Parkinsonian symptoms in DLB may be more closely coupled to neurochemical rather than structural changes in the caudate nucleus, and volumetric MRI analysis of caudate nucleus does not discriminate between patients with DLB, AD, and VaD. PMID- 11861710 TI - A case of paraneoplastic syndrome accompanied by two types of cancer. AB - A case of paraneoplastic syndrome accompanied by two types of cancer is reported. The patient was a 62 year old man who progressively developed cerebellar ataxia, especially an abnormal gait. The anti-Hu antibody titre was high. A small tumour was detected in the middle lobe of the right lung and was surgically treated. The histology was adenocarcinoma. After lobectomy, however, the ataxia deteriorated, and plasma exchange, 250 ml/kg/day, was conducted for 6 days. After plasma exchange, the anti-Hu antibody titre decreased and the ataxia temporarily ceased to progress. A week after the last plasma exchange, a mass appeared in the anterior cervical region and rapidly increased in size. The biopsy of the neck tumour disclosed a small cell carcinoma. Five months later small cell carcinoma appeared in the left lung. This case shows the importance of searching for small cell carcinoma when anti-Hu antibodies are detected. It is assumed that plasma exchange removed not only a pathogenic factor of ataxia but also a factor which inhibited the growth of the small cell carcinoma. It is recommended that plasmapheresis should be performed with caution in paraneoplastic syndrome when the origin of a tumour is obscure. PMID- 11861711 TI - Multiple cavernous haemangioma showing marked calcification on cranial radiography. PMID- 11861712 TI - Brain abscess formed in the cavum septi pellucidi. PMID- 11861713 TI - Bromide, the first effective antiepileptic agent. PMID- 11861714 TI - T cell recognition of a non-protein antigen preparation of Campylobacter jejuni in patients with Guillain-Barre syndrome. PMID- 11861715 TI - Changes in excitability of motor cortex in severe hepatic failure. PMID- 11861716 TI - Amnesia for childhood in patients with unexplained neurological symptoms. PMID- 11861717 TI - Anti-GQ1b IgG antibody syndrome: clinical and immunological range. PMID- 11861718 TI - Parkin gene related neuronal multisystem disorder. PMID- 11861719 TI - Obsessive-compulsive phenomenon and Parkinson's disease. PMID- 11861720 TI - Limited open repair of Achilles tendon ruptures: a technique with a new instrument and findings of a prospective multicenter study. AB - BACKGROUND: Controversy persists regarding the ideal surgical technique for repair of a ruptured Achilles tendon. We propose a limited open procedure with use of an instrument that provides the advantage of an open repair but avoids the soft-tissue problems with which open repair has been associated. METHODS: We first performed a cadaver study in order to develop an instrument and a technique for a limited open repair and then, using this procedure in conjunction with an early functional rehabilitation protocol, we began a prospective multicenter study. We are reporting on the first eighty-seven patients consecutively treated with the new instrument and followed for an average of twenty-six months (range, eighteen to forty-two months). All patients were assessed clinically and with an enhanced American Orthopaedic Foot and Ankle Society (AOFAS) rating score. In addition, all fifty patients who had been followed for at least twenty-four months were further evaluated with isokinetic dynamometry. RESULTS: Four patients were lost to follow-up and one patient died, which left eighty-two patients for evaluation. There were no problems with wound-healing, and there were no infections. No patient noted a sensory disturbance in the sural nerve distribution. All patients returned to their previous professional or sporting activities. The mean AOFAS score was 96 points (range, 85 to 100 points). Isokinetic dynamometry showed no significant difference in strength between the injured and uninjured limbs of the fifty patients who were tested. Complications occurred in three patients. Two of them were noncompliant and removed the orthosis, so that the repair was disrupted by a new injury within the first three weeks postoperatively. One patient fell twelve weeks after the surgery and sustained a rerupture. All three new injuries were repaired with an open surgical procedure. CONCLUSIONS: This new procedure allows the surgeon to precisely visualize and control the tendon ends while avoiding excessive dissection and disturbance of local vascularity and minimizing nerve and wound-healing problems. Such a technique, along with an early functional rehabilitation program, allowed us to achieve a high rate of successful results with minimal morbidity. PMID- 11861721 TI - Twenty-five-year survivorship of two thousand consecutive primary Charnley total hip replacements: factors affecting survivorship of acetabular and femoral components. AB - BACKGROUND: Charnley total hip arthroplasty has been demonstrated to provide good clinical results and a high rate of implant survivorship for twenty years and longer. Most long-term series are not large enough to stratify the many demographic factors that influence implant survivorship. The purpose of this study was to analyze the effects of demographic factors and diagnoses on the long term survivorship of the acetabular and femoral components used in Charnley total hip arthroplasty. METHODS: Two thousand primary Charnley total hip arthroplasties (1689 patients) were performed at one institution from 1969 to 1971. Patients were contacted at five-year intervals after the arthroplasty. Twenty-five years after the surgery, 1228 patients had died and 461 patients were living. Hips that had not had a reoperation, revision or removal of a component for any reason, or revision or removal for aseptic loosening were considered to have survived. Survivorship data were calculated with use of the method of Kaplan and Meier. Patients were stratified by age, gender, and underlying diagnosis to determine the influence of these factors on implant survivorship. RESULTS: The twenty-five year rates of survivorship free of reoperation, free of revision or removal of the implant for any reason, and free of revision or removal for aseptic loosening were 77.5%, 80.9% and 86.5%, respectively. The twenty-five-year survivorship free of revision for aseptic loosening was poorer for each decade earlier in life at which the procedure was performed; this survivorship ranged from 68.7% for patients who were less than forty years of age to 100% for patients who were eighty years of age or older. Men had a twofold higher rate of revision for aseptic loosening than did women. CONCLUSIONS: Age, gender, and underlying diagnosis all affected the likelihood of long-term survivorship of the acetabular and femoral components used in Charnley total hip arthroplasty. PMID- 11861722 TI - Salter innominate osteotomy for the treatment of developmental dysplasia of the hip in children: results of seventy-three consecutive osteotomies after twenty six to thirty-five years of follow-up. AB - BACKGROUND: Reorientation of the acetabulum may be necessary in the treatment of an unstable hip in children with developmental dysplasia of the hip. In 1961, Salter described the innominate osteotomy for stabilizing the reduced hip in the position of function by redirection of the acetabulum as one piece. In the present study, we describe our long-term results with this procedure. METHODS: We reviewed the cases of sixty-one patients who had seventy-three Salter innominate osteotomies. At the time of the operation, the mean age of the patients was 4.1 years (range, 1.3 to 8.8 years). Radiographs made preoperatively, postoperatively, and at the time of the most recent follow-up visit were evaluated. Clinical evaluation was performed with use of the Merle d'Aubigne and Postel system as well as the Harris hip score. RESULTS: The mean duration of follow-up was 30.9 years (range, 26.2 to 35.4 years). There were seven true revisions (one acetabuloplasty, one triple osteotomy, and five total hip arthroplasties). With true revision as the end point, the cumulative survival rate at 35.3 years was 0.90. Fifteen of the seventy-three hips were considered a failure, which was defined as a revision or a Harris hip score of <70 points and/or a Merle d'Aubigne and Postel score of <13 points. The long-term clinical outcome was significantly influenced by the grade of dislocation on the radiographs made at the first examination (p = 0.0388) and on those made immediately preoperatively (p < 0.0001), the postoperative summarized hip factor (the radiographic grade of dysplasia) (p = 0.0002), the preoperative (p = 0.0392) and postoperative (p = 0.0072) grades of avascular necrosis of the femoral head, and the technique of reduction (p < 0.0001). CONCLUSIONS: When an acetabulum can be most closely restored to a normal configuration without the development of avascular necrosis, good long-term results (lasting for more than thirty years) can be expected. When open reduction is necessary, it is preferable to perform it separately prior to the Salter innominate osteotomy. The grade of dislocation at the time of the first examination and immediately preoperatively, the grade of avascular necrosis of the femoral head, and the adequacy of surgical correction are important prognostic factors for the long-term clinical result. PMID- 11861723 TI - Patellar resurfacing in total knee arthroplasty: a prospective, randomized trial. AB - BACKGROUND: The management of the patella in total knee arthroplasty is still problematic. We aimed to identify differences in the clinical outcome of total knee arthroplasty according to whether or not patellar resurfacing had been performed in a prospective, randomized study of 220 osteoarthritic knees. METHODS: Two hundred and twenty total knee arthroplasties in 201 patients were randomly assigned to be performed with either resurfacing or retention of the patella, and the results were followed for a mean of forty-eight months (range, thirty-six to seventy-nine months) in a double-blind (both patient and clinical evaluator blinded), prospective study. Evaluation was performed annually by an independent observer and consisted of assessment with the Knee Society clinical rating system, specific evaluation of anterior knee pain, a stair-climbing test, and radiographic examination. RESULTS: Fifteen (12%) of the 128 knees without patellar resurfacing and nine (10%) of the ninety-two knees with patellar resurfacing underwent a revision or another type of reoperation related to the patellofemoral articulation. This difference was not significant (chi square with one degree of freedom = 0.206, p = 0.650). At the time of the latest follow-up, there was a significantly higher incidence of anterior pain (chi square with one degree of freedom = 5.757, p = 0.016) in the knees that had not had patellar resurfacing. CONCLUSIONS: Patients who underwent patellar resurfacing had superior clinical results in terms of anterior knee pain and stair descent. However, anterior knee pain still occurred in patients with patellar resurfacing, and nine (10%) of the ninety-two patients in that group underwent a revision or another type of reoperation involving the patellofemoral joint. Weight but not body mass index was associated with the development of anterior knee pain in the patients without patellar resurfacing, a finding that suggests that patellofemoral dysfunction may be a function of joint loading rather than obesity. PMID- 11861724 TI - Sonography for monitoring closed reduction of displaced extra-articular distal radial fractures. AB - BACKGROUND: Closed reduction and cast immobilization are employed in the primary treatment of most distal radial fractures, and conventional radiographic techniques have been essential and effective in monitoring these reductions. Radiation-free ultrasonography, however, can provide both real-time and dynamic multiple-plane images with a small and simple-to-use transducer that can be operated with only one hand. We therefore wanted to see if the real-time and dynamic multiple-plane observation capabilities of ultrasonography would allow an orthopaedic surgeon to perform a closed reduction without multiple attempts, as are frequently required when only conventional radiographic techniques are used. METHODS: Sonographically guided closed reduction was performed in twenty-seven consecutive wrists with an acute distal radial fracture. The efficacy of this method was evaluated and compared with that of conventional radiographic techniques. RESULTS: The sonographic images delineated the fractures as accurately as did the conventional radiographs. All parameters measured on the sonograms and radiographs showed substantial restoration of anatomic alignment after reduction, and all measurements were similar on the two types of images. CONCLUSIONS: Sonographically guided monitoring compared well with conventional radiographic techniques during closed reduction of extra-articular distal radial fractures. Sonography is an accurate, simple, and radiation-free tool that provides the substantial benefits of dynamic multiple-plane and real-time observation. PMID- 11861725 TI - Long-term results of total hip arthroplasty with a cemented custom-designed swan neck femoral component for congenital dislocation or severe dysplasia: a follow up note. AB - BACKGROUND: This follow-up study updates the results in a consecutive series of nineteen cemented total hip replacements with a swan-neck femoral component in patients with congenital dislocation or severe hip dysplasia. The series was previously reported on in 1993. METHODS: The patients were petite, with an average height of 152 cm and an average weight of 50 kg, and the femoral canals could not accommodate an off-the-shelf femoral component. Sixteen of the nineteen hips were available for follow-up at an average of 13.3 years (range, eleven to twenty years). Fourteen hips had up-to-date clinical and radiographic examinations. RESULTS: At the time of the latest follow-up, thirteen hips were rated as excellent; two, as good; and one, as a failure because of loosening of both components requiring revision eleven years after the index operation. Another hip required acetabular revision because of loosening fifteen years after the index operation. The rates of femoral and acetabular component revision were 6% and 12.5%, respectively. Radiographic analysis demonstrated that no femoral component was loose. One cup was definitely loose at 19.5 years, and three cups were possibly loose at an average of fourteen years. The radiographic rate of acetabular loosening was 33%. The total rate of cup failure (radiographic loosening and revision) was 43%. CONCLUSIONS: The excellent clinical and radiographic results associated with the swan-neck femoral component, and the 94% rate of survival, at an average of 13.3 years (range, eleven to twenty years) indicate that the biomechanical objectives of this custom-designed prosthesis for patients with congenital dislocation or severe hip dysplasia were met. On the basis of this favorable long-term experience, we still use this prosthesis when the anatomic abnormality cannot be adequately addressed by use of a commercially available prosthetic component. PMID- 11861726 TI - The American academy of orthopaedic surgeons outcomes instruments: normative values from the general population. AB - BACKGROUND: The collection of population-based normative data is a necessary step in the process of standardization of eleven American Academy of Orthopaedic Surgeons (AAOS) musculoskeletal outcomes measures. These data serve as comparative normative scores with which to assess the effectiveness of treatment regimens in clinical practice settings and to study the clinical outcomes of treatment in musculoskeletal research. METHODS: With use of a panel mail methodology, self-reported data on the eleven AAOS musculoskeletal outcomes measures were collected from the general population of the United States. RESULTS: The overall response rate of 67.4% for the various surveys met study expectations. For the eleven measures, the range of the confidence intervals for the surveys was +/-1.6% to +/-2.3%, exceeding the +/-3% set a priori. With use of the Multitrait/Multi-Item Analysis Program, all of the scales within each of eleven measures exhibited high internal reliability as well as discriminant and convergent validity. Items within each of the scales contributed roughly equal proportions of information to the total scale scores. CONCLUSIONS: All eleven instruments met study expectations for providing reliable and valid normative data for use in clinical and research settings. PMID- 11861727 TI - Preoperative hemoglobin levels and the need for transfusion after prosthetic hip and knee surgery: analysis of predictive factors. AB - BACKGROUND: Several studies have established a relationship between the preoperative hemoglobin level and the need for postoperative blood transfusion. We analyzed the relationship between preoperative hemoglobin levels, as well as other factors such as age, gender, weight, height, type and duration of the total joint replacement surgery, and the need for postoperative blood transfusion. METHODS: A retrospective study of 296 patients treated with 370 procedures (209 total hip arthroplasties [56.5%] and 161 total knee arthroplasties [43.5%]) from 1994 to 1998 was carried out. A univariate analysis was performed to establish the relationship between all independent variables and the need for postoperative transfusion. Variables that were determined to have a significant relationship were included in a multivariate analysis. RESULTS: The univariate analysis revealed a significant relationship between the need for postoperative blood transfusion and preoperative hemoglobin levels (p = 0.0001), duration of surgery (p = 0.0001), weight (p = 0.002), height (p = 0.019), and gender (p = 0.0056). However, the multivariate analysis identified a significant relationship only between the need for transfusion and the preoperative hemoglobin level (p = 0.0001) and weight (p = 0.011); height (p = 0.776) and gender (p = 0.122) were discounted as significant factors. Patients with a preoperative hemoglobin level of <130 g/L had a four times greater risk of having a transfusion than did those with a hemoglobin level between 130 and 150 g/L and a 15.3 times greater risk than did those with a hemoglobin level of >150 g/L. CONCLUSIONS: The preoperative hemoglobin level (p = 0.0001) and weight of the patient (p = 0.011) were shown to predict the need for blood transfusion after hip and knee replacement. PMID- 11861728 TI - The role of flexor tenosynovectomy in the operative treatment of carpal tunnel syndrome. AB - BACKGROUND: We conducted a prospective, randomized study to evaluate the effect of flexor tenosynovectomy as an adjunct to open carpal tunnel release for the treatment of idiopathic carpal tunnel syndrome and reviewed the histological characteristics of the flexor tenosynovium to identify possible correlations between histopathology and symptoms. METHODS: Eighty-eight wrists in eighty-seven patients with idiopathic carpal tunnel syndrome were randomized to open carpal tunnel release with or without flexor tenosynovectomy. A validated self administered questionnaire for the assessment of symptom severity and functional status was completed both before and after the operation to assess patient outcome. The study group included fifteen men and seventy-two women with a mean age of fifty-eight years. All patients were followed for a minimum of twelve months after the operation. Intraoperatively, the tenosynovium of all patients was graded on the basis of its gross appearance. Half of the wrists were then treated with a flexor tenosynovectomy through the operative incision, and the tenosynovium was graded histologically. Correlations were sought between the gross appearance of the tenosynovium and the preoperative and postoperative symptoms and functional status, between the histologic appearance of the tenosynovium and the preoperative and postoperative symptoms and functional status, and between the gross and the histologic findings. RESULTS: After the operation, both groups improved significantly with respect to symptom severity and functional status (paired t test), with no significant difference between the groups (unpaired t test). No significant correlation was found between the gross appearance of the tenosynovium and the preoperative or postoperative symptoms and functional status, between the histologic appearance of the tenosynovium and the preoperative or postoperative symptoms and functional status, or between the gross and the histologic findings. CONCLUSIONS: We observed neither an added benefit nor an increased rate of morbidity in association with the performance of a flexor tenosynovectomy at the time of carpal tunnel release. We identified no clinical correlations that might predict which individuals would benefit from flexor tenosynovectomy on the basis of either the gross (intraoperative) or histologic evaluation of the flexor tenosynovium. Our findings suggest that routine flexor tenosynovectomy offers no benefit compared with sectioning of the transverse carpal ligament alone for the treatment of idiopathic carpal tunnel syndrome. PMID- 11861729 TI - Alendronate does not inhibit early bone apposition to hydroxyapatite-coated total joint implants: a preliminary study. AB - BACKGROUND: Alendronate is a pyrophosphate analogue of bisphosphonate that has been shown to inhibit osteoclastic bone resorption. Bone formation and remodeling are necessary to establish initial fixation of uncemented implants, especially those coated with a bioactive surface such as hydroxyapatite. Because the process of bone-remodeling that culminates in new-bone formation is thought to be initiated by osteoclastic bone resorption, it is appropriate to test the influence of osteoclast-inhibiting medications on bone apposition to hydroxyapatite-coated implants. METHODS: Twelve dogs underwent staged bilateral total hip arthroplasty, with twenty weeks between the first and second operations, with use of a titanium-alloy femoral stem that had a proximal macrotextured surface and a plasma-sprayed hydroxyapatite coating. Six of the dogs received oral alendronate therapy from the time of the surgery until they were killed; the other six dogs were untreated controls. The animals were killed four weeks after the second operation. Sections from matched implant sites (proximal, middle, and distal) were histologically analyzed. The linear extent of bone apposition, the linear extent and the thickness of the hydroxyapatite coating, and the total amount of cortical and trabecular bone were measured with the use of an interactive image analysis system. RESULTS: There were no significant differences in radiographic or histologic findings between the two groups at either four or twenty-four weeks. Although the extent of the hydroxyapatite coating decreased significantly with time in both groups (p < 0.01), we identified no significant influence of alendronate on the extent of bone apposition, the extent or thickness of the hydroxyapatite coating, or the cortical or trabecular bone area around the implants. CONCLUSIONS: Many patients who are receiving alendronate for osteoporosis or other disorders may also be candidates for cementless total joint arthroplasty. Although bone formation is generally thought to be initiated by and coupled with bone resorption, our results suggest that alendronate has no discernible effect on the initial fixation of or the short-term bone-remodeling around hydroxyapatite-coated femoral total joint implants. PMID- 11861730 TI - Effects of alendronate on particle-induced osteolysis in a rat model. AB - BACKGROUND: Particle-induced osteolysis is currently a major problem affecting the long-term survivorship of total joint replacements. Alendronate is a third generation bisphosphonate that blocks osteoclastic bone resorption. The objective of this study was to determine whether alendronate could prevent particle-induced osteolysis or restore (reverse) bone loss in established osteolysis. METHODS: A rat model of particle-induced osteolysis was used. A specially designed polyethylene implant was placed in the proximal part of the right tibia of seventy-two animals. Following four weeks of healing, the animals were randomized into control groups, a prevention group, or a treatment group. In the prevention group, animals received intra-articular injections of high-density polyethylene particles (mean size, 2 m; all <10 m) at four, six, and eight weeks postoperatively. Alendronate (0.01 mg/kg/day) was administered concomitantly through an implantable pump from the fourth week through the tenth week. In the treatment group, animals were also exposed to polyethylene particles at four, six, and eight weeks, to establish bone loss, but they received alendronate subsequently, from the tenth week through the sixteenth week, to treat the bone loss. Positive (particle-only) and negative (saline-solution-only) control groups were assessed as well. Tissues were harvested at ten weeks in the prevention group and at sixteen weeks in the treatment group. Histological analyses and histomorphometric determinations of the periprosthetic bone volume were carried out. RESULTS: Histological examination showed a rim of new bone (neocortex) around the implant in the untreated and saline-solution-treated control animals (no polyethylene particles). Treatment with saline solution (no polyethylene particles) did not affect periprosthetic bone. Animals exposed to polyethylene particles had bone loss. In those that received alendronate, the bone loss was either prevented or reversed, and the quantity of neocortical and trabecular bone was increased compared with that of the controls. Alendronate effectively preserved periprosthetic bone in both the prevention and treatment groups. In the prevention arm, the mean periprosthetic bone volume of the neocortex and the surrounding trabecular bone, as determined with histomorphometry, was 21.5% +/- 6.5% in the saline-solution-treated controls (no particles), 13.1% +/- 5.9% in the particle-treated animals, and 32.6% +/- 6.4% in the alendronate-treated animals (p < 0.001). In the treatment arm, the mean periprosthetic bone volume was 27.2% +/- 5.6% in the saline-solution-treated controls, 17.7% +/- 6.2% in the particle-treated animals, and 30.2% +/- 5.9% in the alendronate-treated animals (p = 0.002). CONCLUSIONS: In our model, the intra-articular injection of polyethylene particles caused substantial bone loss around a loaded implant. Alendronate effectively prevented and treated the particle-induced periprosthetic bone loss. PMID- 11861731 TI - The fate of stable femoral components retained during isolated acetabular revision: a six-to-twelve-year follow-up study. AB - BACKGROUND: The decision as to whether to revise a well-fixed femoral component in hips requiring isolated acetabular revision is challenging. The purpose of the present study was to determine the long-term results of, and the complications associated with, retention of a stable and well-fixed femoral component during isolated acetabular revision. METHODS: We retrospectively reviewed the clinical and radiographic results for thirty-one patients (thirty-two hips) who underwent isolated revision acetabuloplasty without removal of a well-fixed femoral component. The reason for acetabular revision was aseptic loosening in thirty-one hips and malposition in one hip. Of the thirty-two femoral components, twenty-one were cemented and eleven were cementless. The average duration of follow-up from the time of the index revision was 8.1 years (range, 6.4 to 12.5 years), and the average duration of total service of the femoral component was seventeen years (range, seven to twenty-five years) from time of the initial implantation. The average age of the patients at the time of the index revision was sixty-six years (range, twenty-nine to eighty-seven years). RESULTS: Thirty-one (97%) of the primary femoral components were judged to be stable and well fixed at the latest follow-up evaluation. One femoral component (3%) was revised because of aseptic loosening, eight years after the index acetabular revision and seventeen years after the initial total hip arthroplasty. Radiographic evaluation of the thirty one femoral components that were not revised demonstrated no evidence of loosening or subsidence. There were no dislocations, nerve palsies, or intraoperative fractures associated with retention of the femoral component. Twenty-seven (84%) of the acetabular components were judged to be stable at the latest follow-up evaluation. CONCLUSION: In hips treated with isolated acetabular revision, a well-fixed femoral component can be retained successfully without adversely affecting the acetabular exposure; the placement, position, or stability of the acetabular component; or the ability to restore bone stock. The data from the present study support the decision to retain a well-fixed femoral component when the acetabular component needs to be revised. PMID- 11861732 TI - Jumbo femoral head for the treatment of recurrent dislocation following total hip replacement. AB - BACKGROUND: The purpose of this study was to assess the results of the use of a jumbo femoral head to restore stability in hips that had sustained recurrent dislocations after total hip replacement. METHODS: Twelve hips in twelve patients who had had multiple hip operations and recurrent instability of the hip underwent a total hip replacement with use of a femoral head with an average diameter of 44 mm (range, 40 to 50 mm). The average age of the patients was fifty nine years (range, twenty-nine to eighty-four years). The twelve patients had had an average of four previous operations (range, one to eight operations) and seven dislocations (range, two to twenty dislocations). A bipolar head was used in ten hips that had a femoral stem with a fixed (non-modular) head, and a modular head (unipolar) was used in two hips. (One hip was first treated with a bipolar head and then with a unipolar head.) RESULTS: One patient died of unrelated causes fourteen months postoperatively. The hip had remained stable until the time of death. After an average duration of follow-up of 6.5 years (range, 3.2 to 11.8 years), ten of the remaining eleven hips had had no additional episodes of instability. One hip dislocated within one week after the revision, necessitating revision surgery to reposition the acetabular component. This hip was found to be stable at the time of follow-up 7.6 years after the revision. There were four other reoperations: one was done because of a fracture of the polyethylene; one, because of entrapment of cement within the articulation; one, because of pain related to loosening of the femoral stem; and one, because of late hematogenous infection. The preoperative and postoperative University of California at Los Angeles hip scores for the series were, respectively, 7 and 9 points for pain, 5 and 7 points for walking, 4 and 6 points for function, and 3 and 5 points for activity. CONCLUSIONS: A jumbo-diameter femoral head provided stability and improved function without compromising range of motion in patients with recurrent dislocations following total hip arthroplasty. PMID- 11861733 TI - Gait characteristics after limb-sparing surgery with sciatic nerve resection: a report of two cases. PMID- 11861734 TI - Histological findings in a proximal femoral structural allograft ten years following revision total hip arthroplasty: a case report. PMID- 11861735 TI - Flexion osteotomy of the metacarpal neck: a treatment method for avascular necrosis of the head of the third metacarpal: a case report. PMID- 11861736 TI - Congenital osseous anomalies of the upper and lower cervical spine in children. PMID- 11861738 TI - Continuing concerns, new challenges, and next steps in physician-patient communication. PMID- 11861737 TI - Congenital clubfoot. PMID- 11861739 TI - Treatment of scoliosis: correction and internal fixation by spine instrumentation. June 1962. PMID- 11861740 TI - Limb-lengthening versus amputation for fibular hemimelia. PMID- 11861741 TI - Methods of cast immobilization for nondisplaced scaphoid fractures and the evaluation of fracture union. PMID- 11861743 TI - The semantics of terminology: distinguishing Arnold-Chiari malformations from Chiari malformations. PMID- 11861742 TI - Rupture of the Achilles tendon in athletes: do synthetic grass fields play a part? PMID- 11861745 TI - Three surgical methods of anterior cruciate ligament reconstruction were equally effective. PMID- 11861747 TI - Surgical treatment of acute rupture of the tendo Achillis led to fewer reruptures and better patient-generated ratings than did nonsurgical treatment. PMID- 11861749 TI - The subvastus approach for total knee arthroplasty resulted in better short-term outcomes than did the parapatellar approach. PMID- 11861750 TI - What's new in hand surgery. PMID- 11861751 TI - How to impress without even trying. By Caveman. PMID- 11861753 TI - The role of beta-arrestins in the termination and transduction of G-protein coupled receptor signals. AB - beta-Arrestins are versatile adapter proteins that form complexes with most G protein-coupled receptors (GPCRs) following agonist binding and phosphorylation of receptors by G-protein-coupled receptor kinases (GRKs). They play a central role in the interrelated processes of homologous desensitization and GPCR sequestration, which lead to the termination of G protein activation. beta arrestin binding to GPCRs both uncouples receptors from heterotrimeric G proteins and targets them to clathrin-coated pits for endocytosis. Recent data suggest that beta-arrestins also function as GPCR signal transducers. They can form complexes with several signaling proteins, including Src family tyrosine kinases and components of the ERK1/2 and JNK3 MAP kinase cascades. By recruiting these kinases to agonist-occupied GPCRs, beta-arrestins confer distinct signaling activities upon the receptor. beta-arrestin-Src complexes have been proposed to modulate GPCR endocytosis, to trigger ERK1/2 activation and to mediate neutrophil degranulation. By acting as scaffolds for the ERK1/2 and JNK3 cascades, beta arrestins both facilitate GPCR-stimulated MAP kinase activation and target active MAP kinases to specific locations within the cell. Thus, their binding to GPCRs might initiate a second wave of signaling and represent a novel mechanism of GPCR signal transduction. PMID- 11861754 TI - The COP9 signalosome: at the interface between signal transduction and ubiquitin dependent proteolysis. AB - Recently the COP9 signalosome (CSN) has become a focus of interest for many researchers, because of its function at the interface between signal transduction and ubiquitin-dependent proteolysis. It is required for the proper progression of the cell cycle in Schizosaccharomyces pombe and is essential for development in plants and Drosophila. However, its function in mammalian cells remains obscure. Although the CSN shares structural similarities with the 26S proteasome lid complex (LID), its functions seem to be different from that of the LID. A variety of CSN-specific protein-protein interactions have been described in mammalian cells. However, it is currently unclear how many reflect true functions of the complex. Two activities associated with the CSN have been identified so far: a protein kinase and a deneddylase. The CSN-associated kinase phosphorylates transcription factors, which determines their stability towards the ubiquitin system. The associated deneddylase regulates the activity of specific SCF E3 ubiquitin ligases. The CSN thus appears to be a platform connecting signalling with proteolysis. PMID- 11861756 TI - Menadione-induced apoptosis: roles of cytosolic Ca(2+) elevations and the mitochondrial permeability transition pore. AB - In normal pancreatic acinar cells, the oxidant menadione evokes repetitive cytosolic Ca(2+) spikes, partial mitochondrial depolarisation, cytochrome c release and apoptosis. The physiological agonists acetylcholine and cholecystokinin also evoke cytosolic Ca(2+) spikes but do not depolarise mitochondria and fail to induce apoptosis. Ca(2+) spikes induced by low agonist concentrations are confined to the apical secretory pole of the cell by the buffering action of perigranular mitochondria. Menadione prevents mitochondrial Ca(2+) uptake, which permits rapid spread of Ca(2+) throughout the cell. Menadione-induced mitochondrial depolarisation is due to induction of the permeability transition pore. Blockade of the permeability transition pore with bongkrekic acid prevents activation of caspase 9 and 3. In contrast, the combination of antimycin A and acetylcholine does not cause apoptosis but elicits a global cytosolic Ca(2+) rise and mitochondrial depolarisation without induction of the permeability transition pore. Increasing the cytosolic Ca(2+) buffering power by BAPTA prevents cytosolic Ca(2+) spiking, blocks the menadione-elicited mitochondrial depolarisation and blocks menadione-induced apoptosis. These results suggest a twin-track model in which both intracellular release of Ca(2+) and induction of the permeability transition pore are required for initiation of apoptosis. PMID- 11861755 TI - Sites of Ca(2+) wave initiation move with caveolae to the trailing edge of migrating cells. AB - The caveola is a membrane domain that compartmentalizes signal transduction at the cell surface. Normally in endothelial cells, groups of caveolae are found clustered along stress fibers or at the lateral margins in all regions of the cell. Subsets of these clusters appear to contain the signaling machinery for initiating Ca(2+) wave formation. Here we report that induction of cell migration, either by wounding a cell monolayer or by exposing cells to laminar shear stress, causes caveolae to move to the trailing edge of the cell. Concomitant with the relocation of the caveolae, sites of Ca(2+) wave initiation move to the same location. In as much as the relocated caveolae contain elements of the signaling machinery required for ATP-stimulated release of Ca(2+) from the ER, these results suggest that caveolae function as containers that carry this machinery to different cellular locations. PMID- 11861757 TI - Gelsolin--evidence for a role in turnover of junction-related actin filaments in Sertoli cells. AB - The gelsolin-phosphoinositide pathway may be part of the normal mechanism by which Sertoli cells regulate sperm release and turnover of the blood-testis barrier. The intercellular adhesion complexes (ectoplasmic specializations) involved with these two processes are tripartite structures consisting of the plasma membrane, a layer of actin filaments and a cistern of endoplasmic reticulum. Gelsolin is concentrated in these adhesion complexes. In addition, phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P(2)) and phosphoinositide specific phospholipase C are found in the structures. Treatment of isolated spermatid/junction complexes with exogenous phosphoinositide-specific phospholipase C, or with a synthetic peptide consisting of the PtdIns(4,5)P(2) binding region of gelsolin, results in the release of gelsolin and loss of actin from the adhesion complexes. We present a model for the disassembly of the actin layer of the adhesion complex that involves the hydrolysis of PtdIns(4,5)P(2) resulting in the release of gelsolin within the plaque. Further, we speculate that the hydrolysis of PtdIns(4,5)P(2) may result in a local Ca(2+) surge via the action of inositol triphosphate on junctional endoplasmic reticulum. This Ca(2+) surge would facilitate the actin severing function of gelsolin within the adhesion complex. PMID- 11861758 TI - Merotelic kinetochore orientation versus chromosome mono-orientation in the origin of lagging chromosomes in human primary cells. AB - Defects in chromosome segregation play a critical role in producing genomic instability and aneuploidy, which are associated with congenital diseases and carcinogenesis. We recently provided evidence from immunofluorescence and electron microscopy studies that merotelic kinetochore orientation is a major mechanism for lagging chromosomes during mitosis in PtK1 cells. Here we investigate whether human primary fibroblasts exhibit similar errors in chromosome segregation and if at least part of lagging chromosomes may arise in cells entering anaphase in the presence of mono-oriented chromosomes. By using in situ hybridization with alphoid probes to chromosome 7 and 11 we showed that loss of a single sister is much more frequent than loss of both sisters from the same chromosome in anatelophases from human primary fibroblasts released from a nocodazole-induced mitotic arrest, as predicted from merotelic orientation of single kinetochores. Furthermore, the lagging of pairs of separated sisters was higher than expected from random chance indicating that merotelic orientation of one sister may promote merotelic orientation of the other. Kinetochores of lagging chromosomes in anaphase human cells were found to be devoid of the mitotic checkpoint phosphoepitopes recognized by the 3F3/2 antibody, suggesting that they attached kinetochore microtubules prior to anaphase onset. Live cell imaging of H2B histone-GFP-transfected cells showed that cells with mono-oriented chromosomes never enter anaphase and that lagging chromosomes appear during anaphase after chromosome alignment occurs during metaphase. Thus, our results demonstrate that the mitotic checkpoint efficiently prevents the possible aneuploid burden due to mono-oriented chromosomes and that merotelic kinetochore orientation is a major limitation for accurate chromosome segregation and a potentially important mechanism of aneuploidy in human cells. PMID- 11861759 TI - Pax3 induces cell aggregation and regulates phenotypic mesenchymal-epithelial interconversion. AB - Paired box-containing transcription factors play fundamental roles in pattern formation during embryonic development of diverse organisms ranging from Drosophila to mammals. Although mutations to Pax3 and other Pax-family genes in both mice and humans result in numerous tissue-specific morphological defects, little is known about the cellular processes that Pax genes regulate. We show that extopic Pax3 expression in two distinct phenotypically mesenchymal mammalian cell lines induces the formation of multi-layered condensed cell aggregates with epithelial characteristics. For one of these lines, we showed further that Pax3 induced cell aggregation is accompanied by specific morphological changes, including a significant reduction in cell size, altered cell shape and dramatic alterations to both membrane and cytoskeleton architecture. In addition to mediating a phenotypic mesenchymal-to-epithelial transition, Pax3 also establishes the conditions in these cells for a subsequent hepatocyte growth factor/scatter factor (HGF/SF)-induced phenotypic epithelial-to-mesenchymal transition. Thus, our data show a novel morphogenetic activity for Pax3 which, when absent in vivo, is predicted to give rise to the observed structural defects in somites and the neural tube during embryonic development. PMID- 11861761 TI - Integrin-mediated functional polarization of Caco-2 cells through E-cadherin- actin complexes. AB - Enterocyte differentiation is a dynamic process during which reinforcement of cell-cell adhesion favours migration along the crypt-to-villus axis. Functional polarization of Caco-2 cells, the most commonly used model to study intestinal differentiation, is assessed by dome formation and tightness of the monolayer and is under the control of the extracellular matrix (ECM). Furthermore, our biochemical and confocal microscopy data demonstrate that the ECM dramatically reinforces E-cadherin targeting to the upper lateral membrane, formation of the apical actin cytoskeleton and its colocalization with E-cadherin in functional complexes. In our model, these effects were produced by native laminin-5-enriched ECM as well as by type IV collagen or laminin 2, which suggests a common pathway of induction through integrin receptors. Indeed, these effects were antagonized by blocking anti-beta1- and anti-alpha6-integrin antibodies and directly induced by a stimulating anti-beta1-integrin antibody. These results demonstrate that integrin-dependent cell to ECM adhesion reinforces E-cadherin-dependent cell-cell adhesion in Caco-2 cells and further support the notion that enterocyte differentiation is supported by a molecular crosstalk between the two adhesion systems of the cell. PMID- 11861760 TI - Pax3 regulates morphogenetic cell behavior in vitro coincident with activation of a PCP/non-canonical Wnt-signaling cascade. AB - Mutations to Pax3 and other Pax family genes in both mice and humans result in numerous tissue-specific morphological defects. Little is known, however, about the cellular and molecular mechanisms by which Pax genes regulate morphogenesis. We previously showed that Pax3 induces cell aggregation and a mesenchymal-to epithelial transition in Saos-2 cells. We show here that Pax3-induced aggregates arise through the formation of distinct structures involving cell rearrangements and cell behaviors resembling those that occur during gastrulation and neurulation known as convergent extension. During these Pax3-induced processes, Dishevelled and Frizzled are localized to the actin cytoskeleton and both proteins coimmunoprecipitate focal adhesion components from detergent-insoluble cell fractions. We show further that these Pax3-induced cell movements are associated with activation of a Wnt-signaling cascade, resulting in induction and activation of c-Jun-N-terminal kinase/stress activated protein kinase (JNK/SAPK). All of these Wnt-signaling factors exhibit altered subcellular distribution in Pax3-expressing cells. In particular, we show the localization of JNK/SAPK to both the nucleus and to cytoplasmic multi-vesicular structures. These data show that Pax3 regulates morphogenetic cell behavior and that regulation of a conserved, planar cell polarity/noncanonical Wnt-signaling cascade entailing JNK activation is a function of Pax3 activity. PMID- 11861762 TI - Induction of chondrocyte growth arrest by FGF: transcriptional and cytoskeletal alterations. AB - The effect of fibroblast growth factor (FGF) on mature chondrocytes, the cells responsible for axial skeletal development, is growth attenuation rather than stimulation. This singular response has been linked to signaling via FGF receptor 3 (FGFR3), partly because mutations causing chronic FGFR3 activation lead to various human disorders of bone growth. In order to study how FGF inhibits growth, we analyzed its effect on a rat chondrocyte-derived cell line. We show that the FGF-induced growth arrest occurs at the G1 phase, accompanied by profound changes in gene expression and cytoskeletal organization. Within minutes of binding, FGF induces tyrosine kinase activity in the focal substrate adhesions where it colocalizes with vinculin. Upon FGF stimulation, FGFR3 is selectively removed from the focal adhesions, which is followed by their disassembly and disruption of the organized cytoskeleton. Multiple genes are induced following FGF stimulation in chondrocytes, which has been shown by DNA array screening and confirmed for some by immunoblotting. These genes include regulators of cell differentiation and proliferation such as c-jun, JunD, cyclin-D1, NFkappaB1 and of plasma-membrane microdomain morphology, such as ezrin. The transcription factor Id1 is downregulated, consistent with the cells' exit from the mitotic cycle. Moreover, following FGF stimulation, levels of FGFR3 mRNA and protein decline, as does downstream signaling through the MAPK pathway. The importance of this FGFR3-mediated on-off control is illustrated in transgenic mice expressing mutant, hyperactive FGFR3, where abnormally high levels of NFkappaB are expressed throughout their bone growth-plates. A working model is presented of the signaling network involved in regulating FGF-induced chondrocyte differentiation and receptor downregulation. PMID- 11861763 TI - A family of transmembrane microneme proteins of Toxoplasma gondii contain EGF like domains and function as escorters. AB - TgMIC6, TgMIC7, TgMIC8 and TgMIC9 are members of a novel family of transmembrane proteins localized in the micronemes of the protozoan parasite Toxoplasma gondii. These proteins contain multiple epidermal growth factor-like domains, a putative transmembrane spanning domain and a short cytoplasmic tail. Sorting signals to the micronemes are encoded in this short tail. We established previously that TgMIC6 serves as an escorter for two soluble adhesins, TgMIC1 and TgMIC4. Here, we present the characterization of TgMIC6 and three additional members of this family, TgMIC7, -8 and -9. Consistent with having sorting signals localized in its C-terminal tail, TgMIC6 exhibits a classical type I membrane topology during its transport along the secretory pathway and during storage in the micronemes. TgMIC6 is processed at the N-terminus, probably in the trans-Golgi network, and the cleavage site has been precisely mapped. Additionally, like other members of the thrombospondin-related anonymous protein family, TgMIC2, TgMIC6 and TgMIC8 are proteolytically cleaved near their C-terminal domain upon discharge by micronemes. We also provide evidence that TgMIC8 escorts another recently described soluble adhesin, TgMIC3. This suggests that the existence of microneme protein complexes is not an exception but rather the rule. TgMIC6 and TgMIC8 are expressed in the rapidly dividing tachyzoites, while TgMIC7 and TgMIC9 genes are predominantly expressed in bradyzoites, where they presumably also serve as escorters. PMID- 11861764 TI - Survivin exists in immunochemically distinct subcellular pools and is involved in spindle microtubule function. AB - Survivin is a member of the inhibitor of apoptosis gene family that has been implicated in both apoptosis inhibition and regulation of mitosis. However, the subcellular distribution of survivin has been controversial and variously described as a microtubule-associated protein or chromosomal passenger protein. Here, we show that antibodies directed to the survivin sequence Ala(3)-Ile(19) exclusively recognized a nuclear pool of survivin that segregated with nucleoplasmic proteins, but not with outer nuclear matrix or nuclear matrix proteins. By immunofluorescence, nuclear survivin localized to kinetochores of metaphase chromosomes, and to the central spindle midzone at anaphase. However, antibodies to Cys(57)-Trp(67) identified a cytosolic pool of survivin, which associated with interphase microtubules, centrosomes, spindle poles and mitotic spindle microtubules at metaphase and anaphase. Polyclonal antibodies recognizing survivin epitopes Ala(3)-Ile(19), Met(38)-Thr(48), Pro(47)-Phe(58) and Cys(57) Trp(67) identified both survivin pools within the same mitotic cell. A ratio of approximately 1:6 for nuclear versus cytosolic survivin was obtained by quantitative subcellular fractionation. In synchronized cultures, cytosolic survivin abruptly increased at mitosis, physically associated with p34(cdc2), and was phosphorylated by p34(cdc2) on Thr(34), in vivo. By contrast, nuclear survivin began to accumulate in S phase, was not complexed with p34(cdc2) and was not phosphorylated on Thr(34). Intracellular loading of a polyclonal antibody to survivin caused microtubule defects and resulted in formation of multipolar mitotic spindles, but did not interfere with cytokinesis. These data demonstrate that although both reported localizations of survivin exist in mitotic cells, the preponderant survivin pool is associated with microtubules and participates in the assembly of a bipolar mitotic spindle. PMID- 11861765 TI - Fission yeast Pds5 is required for accurate chromosome segregation and for survival after DNA damage or metaphase arrest. AB - Sister chromatid cohesion, which is established during the S phase of the eukaryotic cell cycle and persists until the onset of anaphase, is essential for the maintenance of genomic integrity. Cohesion requires the multi-protein complex cohesin, as well as a number of accessory proteins including Pds5/BIMD/Spo76. In the budding yeast Saccharomyces cerevisiae Pds5 is an essential protein that localises to chromosomes in a cohesin-dependent manner. Here we describe the characterisation in the fission yeast Schizosaccharomyces pombe of pds5(+), a novel, non-essential orthologue of S. cerevisiae PDS5. The S. pombe Pds5 protein was localised to punctate nuclear foci in a manner that was dependent on the Rad21 cohesin component. This, together with additional genetic evidence, points towards an involvement of S. pombe Pds5 in sister chromatid cohesion. S. pombe pds5 mutants were hypersensitive to DNA damage and to mitotic metaphase delay, but this sensitivity was apparently not due to precocious loss of sister chromatid cohesion. These cells also suffered increased spontaneous chromosome loss and meiotic defects and their viability was dependent on the spindle checkpoint protein Bub1. Thus, while S. pombe Pds5 has an important cohesin related role, this differs significantly from that of the equivalent budding yeast protein. PMID- 11861766 TI - pH-dependent regulation of lysosomal calcium in macrophages. AB - Calcium measurements in acidic vacuolar compartments of living cells are few, primarily because calibration of fluorescent probes for calcium requires knowledge of pH and the pH-dependence of the probe calcium-binding affinities. Here we report pH-corrected measurements of free calcium concentrations in lysosomes of mouse macrophages, using both ratiometric and time-resolved fluorescence microscopy of probes for pH and calcium. Average free calcium concentration in macrophage lysosomes was 4-6x10(-4) M, less than half of the extracellular calcium concentration, but much higher than cytosolic calcium levels. Incubating cells in varying extracellular calcium concentrations did not alter lysosomal pH, and had only a modest effect on lysosomal calcium concentrations, indicating that endocytosis of extracellular fluid provided a small but measurable contribution to lysosomal calcium concentrations. By contrast, increases in lysosomal pH, mediated by either bafilomycin A(1) or ammonium chloride, decreased lysosomal calcium concentrations by several orders of magnitude. Re-acidification of the lysosomes allowed rapid recovery of lysosomal calcium concentrations to higher concentrations. pH-dependent reductions of lysosomal calcium concentrations appeared to result from calcium movement out of lysosomes into cytoplasm, since increases in cytosolic calcium levels could be detected upon lysosome alkalinization. These studies indicate that lysosomal calcium concentration is high and is maintained in part by the proton gradient across lysosomal membranes. Moreover, lysosomes could provide an intracellular source for physiological increases in cytosolic calcium levels. PMID- 11861768 TI - Role of cytoplasmic C-terminal amino acids of membrane proteins in ER export. AB - Export of membrane proteins from the ER is believed to be selective and require transport signals, but the identity of such signals has remained elusive. The recycling type I membrane protein ERGIC-53 carries a C-terminal diphenylalanine motif that is required for efficient ER export. Here we show that this motif can be functionally substituted by a single phenylalanine or tyrosine at position -2, two leucines or isoleucines at position -1 and -2 or a single valine at position 1. These motifs are common among mammalian type I membrane proteins. A single C terminal valine, but none of the other motifs, accelerates transport of inefficiently exported reporter constructs and hence operates as an export signal. The valine signal is position, but not context, dependent. All transport motifs mediate COPII binding in vitro with distinct preferences for the COPII subunits Sec23p, Sec24Bp, Sec24Cp and p125. These results suggest that cytoplasmic C-terminal amino-acid motifs, either alone or in conjunction with other transport determinants, accelerate ER export of numerous type I and probably polytopic membrane proteins by mediating interaction with COPII coat components. PMID- 11861767 TI - The incorporation of fibrinogen into extracellular matrix is dependent on active assembly of a fibronectin matrix. AB - Fibrinogen is a soluble protein produced by hepatocytes and secreted into plasma, where it functions in hemostasis. During inflammation, the hepatic synthesis of fibrinogen is induced 2-10 fold. Recent studies demonstrate that after an inflammatory stimulus, fibrinogen gene expression and protein production is upregulated in lung epithelial cells, where it is secreted basolaterally and consequently deposited into the extracellular matrix in fibrils that extensively colocalize with fibronectin fibrils. In this study, we show that the deposition of fibrinogen into the matrix of fibroblasts occurred rapidly and in a Rho dependent manner in response to serum or lysophosphatidic acid; RhoA GTPase signaling is also required for fibronectin matrix assembly. Using mouse embryonic fibronectin-null cells, we show that incorporation of exogenous fibrinogen into matrix fibrils occurred only in the presence of exogenous fibronectin, which is also assembled into matrix fibrils. Furthermore, treatment of fibroblasts and fibronectin-null cells with an antibody that inhibits fibronectin matrix assembly impaired incorporation of fibrinogen into matrix fibrils. Collectively, these data suggest that incorporation of fibrinogen into the extracellular matrix requires active fibronectin polymer elongation into matrix fibrils. From these data, we hypothesize that fibrinogen deposition rapidly changes the topology of the extracellular matrix to provide a surface for cell migration and matrix remodeling during tissue repair. PMID- 11861769 TI - Human p63RhoGEF, a novel RhoA-specific guanine nucleotide exchange factor, is localized in cardiac sarcomere. AB - The Rho small GTPases are crucial proteins involved in regulation of signal transduction cascades from extracellular stimuli to cell nucleus and cytoskeleton. It has been reported that these GTPases are directly associated with cardiovascular disorders. In this context, we have searched for novel modulators of Rho GTPases, and here we describe p63RhoGEF a new Db1-like guanine nucleotide exchange factor (GEF). P63RhoGEF encodes a 63 kDa protein containing a Db1 homology domain in tandem with a pleckstrin homology domain and is most closely related to the second Rho GEF domain of Trio. Northern blot and in situ analysis have shown that p63RhoGEF is mainly expressed in heart and brain. In vitro guanine nucleotide exchange assays have shown that p63RhoGEF specifically acts on RhoA. Accordingly, p63RhoGEF expression induces RhoA-dependent stress fiber formation in fibroblasts and in H9C2 cardiac myoblasts. Moreover, we show that p63RhoGEF activation of RhoA in intact cells is dependent on the presence of the PH domain. Using a specific anti-p63RhoGEF antibody, we have detected the p63RhoGEF protein by immunocytochemistry in human heart and brain tissue sections. Confocal microscopy shows that p63RhoGEF is located in the sarcomeric I band mainly constituted of cardiac sarcomeric actin. Together, these results show that p63RhoGEF is a RhoA-specific GEF that may play a key role in actin cytoskeleton reorganization in different tissues, especially in heart cellular morphology. PMID- 11861770 TI - Actin filament turnover removes bundles from Drosophila bristle cells. AB - Drosophila bristle cells form enormous extensions that are supported by equally impressive scaffolds of modular, polarized and crosslinked actin filament bundles. As the cell matures and support is taken over by the secreted cuticle, the actin scaffold is completely removed. This removal begins during cell elongation and proceeds via an orderly series of steps that operate on each module. Using confocal and electron microscopy, we found that the approximately 500-filament modules are fractured longitudinally into 25-50-filament subbundles, indicating that module breakdown is the reverse of assembly. Time-lapse confocal analysis of GFP-decorated bundles in live cells showed that modules were shortened by subunit removal from filament barbed ends, again indicating that module breakdown is the reverse of assembly. Module shortening takes place at a fairly slow rate of approximately 1microm/hour, implying that maximally crosslinked modules are not rapidly depolymerized. Barbed-end depolymerization was prevented with jasplakinolide and accelerated with cycloheximide, indicating that barbed-end maintenance requires continuous protein synthesis. Subbundle adhesion was lost in the presence of cytochalasin, indicating that continuous actin polymerization is required. Thus, these polarized actin filament bundles are dynamic structures that require continuous maintenance owing to protein and actin filament turnover. We propose that after cell elongation, maintenance falls behind turnover, resulting in the removal of this modular cytoskeleton. PMID- 11861771 TI - Vid22p, a novel plasma membrane protein, is required for the fructose-1,6 bisphosphatase degradation pathway. AB - Fructose-1,6-bisphosphatase (FBPase), an important enzyme in the gluconeogenic pathway in Saccharomyces cerevisiae, is expressed when cells are grown in media containing a poor carbon source. Following glucose replenishment, FBPase is targeted from the cytosol to intermediate Vid (vacuole import and degradation) vesicles and then to the vacuole for degradation. Recently, several vid mutants that are unable to degrade FBPase in response to glucose were identified. Here, we present VID22, a novel gene involved in FBPase degradation. VID22 encodes a glycosylated integral membrane protein that localizes to the plasma membrane. Newly synthesized Vid22p was found in the cytoplasm and then targeted to the plasma membrane independent of the classical secretory pathway. A null mutation of VID22 failed to degrade FBPase following a glucose shift and accumulated FBPase in the cytosol. Furthermore, the majority of FBPase remained in a proteinase K sensitive compartment in the Deltavid22 mutant, implying that VID22 is involved in FBPase transport from the cytosol to Vid vesicles. By contrast, starvation-induced autophagy and peroxisome degradation were not impaired in the Deltavid22 mutant. This strain also exhibited the proper processing of carboxypeptidase Y and aminopeptidase I in the vacuole. Therefore, Vid22p appears to play a specific role in the FBPase trafficking pathway. PMID- 11861772 TI - Multiple forms of SNARE complexes in exocytosis from chromaffin cells: effects of Ca(2+), MgATP and botulinum toxin type A. AB - The changes that SNAREs undergo during exocytosis were studied in permeabilised chromaffin cells treated with Ca(2+), MgATP or botulinum neurotoxin A. High resolution 2D SDS-PAGE revealed multiple SDS-resistant SNARE complexes having a wide range of sizes and in which SNAP-25 and syntaxin predominate over synaptobrevin. Their formation increased upon Ca(2+)-stimulated exocytosis; notably, the 2D protocol proved much superior to 1D SDS-PAGE for the detection of large complexes and revealed that for forms with relative molecular mass greater than 100,000 stimulated induction was more significant than for smaller species. MgATP enhanced Ca(2+)-triggered catecholamine release but reduced the content of complexes. By contrast, botulinum neurotoxin type A inhibited exocytosis and altered the stoichiometry of the SNAP-25:syntaxin binary association, without lowering its abundance. The individual SNAREs were protected against trypsin proteolysis to varying extents in binary and ternary complexes of different sizes, suggestive of distinct folding intermediates. Our data suggest that Ca(2+) triggers an early stage of SNARE complex formation causing an accumulation of partially folded intermediates, especially of binary forms, as well as their maturation into smaller, more protease resistant states. In addition, botulinum neurotoxin A inhibits exocytosis by perturbing the syntaxin:SNAP-25 ratio in binary intermediates. PMID- 11861773 TI - Excitotoxicity: perspectives based on N-methyl-D-aspartate receptor subtypes. AB - Since excitotoxicity has been implicated in a variety of neuropathological conditions, understanding the pathways involved in this type of cell death is of critical importance to the future clinical treatment of many diseases. The N methyl-D-aspartate (NMDA) receptor has become a primary focus of excitotoxic research because early studies demonstrated that antagonism of this receptor subtype was neuroprotective. However, initial pharmacological agents were not clinically useful due to the adverse effects of complete NMDA receptor blockade. Understanding the biochemical properties of the multitude of NMDA receptor subtypes offers the possibility of developing more effective and clinically useful drugs. With the discovery of the basis of heterogeneity of NMDA receptors through molecular biological approaches, many new potential therapeutic targets have been uncovered, and several model systems have been developed for the study of NMDA receptor-mediated cell death. This review discusses these models and the current understanding of the relationship between NMDA receptor subtypes and excitotoxicity. PMID- 11861774 TI - Structure-functional diversity of human L-type Ca2+ channel: perspectives for new pharmacological targets. AB - The L-type Ca2+ channels mediate depolarization-induced influx of Ca2+ into a wide variety of cells and thus play a central role in triggering cardiac and smooth muscle contraction. Because of this role, clinically important classes of 1,4-dihydropyridine, phenylalkylamine, and benzothiazepine Ca2+ channel blockers were developed as powerful medicines to treat hypertension and angina pectoris. Molecular cloning studies revealed that the channel is subject to extensive structure-functional variability due to alternative splicing. In this review, we will focus on a potentially important role of genetically driven variability of Ca2+ channels in expression regulation and mutations, Ca2+-induced inactivation, and modulation of sensitivity to Ca2+ channel blockers with the perspective for new pharmacological targets. PMID- 11861775 TI - Role of inflammatory mediators in thrombogenesis. AB - The role of inflammation in cardiovascular disease and especially in thrombogenesis has become increasingly recognized as an important component of the overall disease process. Plaque rupture promotes activation of the inflammatory response and increased expression of tissue factor (TF), which in turn acts as one of the major initiators of extrinsic coagulation. It is becoming apparent that the expression of TF on endothelial cells, underlying smooth muscle cells and monocytes is regulated, in part, by proinflammatory cytokines including tumor necrosis factor and IL-1. In addition to initiating coagulation, interaction of TF with the adhesion molecule, P-selectin, has been demonstrated to accelerate the rate and extent of fibrin formation and deposition. P-selectin is expressed on activated platelets and endothelium and serves as the receptor for the endogenous ligand, P-selectin glycoprotein-1 (PSGL-1), expressed on various leukocytic cell types. In addition to mediating transient interactions between endothelial cells and leukocytes, P-selectin has been reported to mediate adherence of platelets to monocytes and neutrophils via specific interaction with PSGL-1. P-selectin is rapidly cleaved off the surface of the platelet membrane and appears in the circulation as a soluble form, which has been reported to be elevated in patients with acute coronary syndromes including unstable angina and non-Q-wave myocardial infarction. This review will focus on the role of cytokines in mediating TF expression and also explore the significance of the relationship between P-selectin and tissue factor in thrombus generation. In addition, possible pharmacological mechanisms to interrupt this disease process will be discussed. PMID- 11861776 TI - Neurotoxic mechanisms by Alzheimer's disease-linked N141I mutant presenilin 2. AB - Although it has been established that oxidative stress mediates cytotoxicity by familial Alzheimer's disease (FAD)-linked mutants of presenilin (PS)1 and that pertussis toxin inhibits cytotoxicity by FAD-linked N141I-PS2, it has not been determined whether oxidative stress is involved in cytotoxicity by N141I-PS2 or which pertussis toxin-sensitive proteins mediate the cytotoxicity. Here we report that low expression of N141I-PS2 caused neuronal cell death, whereas low expression of wild-type PS2 did not. Cytotoxicities by low and high expression of N141I-PS2 occurred through dissimilar mechanisms: the former cytotoxicity was blocked by a cell-permeable caspase inhibitor, and the latter was not. Since both mechanisms were sensitive to a cell-permeable antioxidant, we examined potential sources of reactive oxygen species in each mechanism, and found that the caspase inhibitor-sensitive neurotoxicity by N141I-PS2 was likely through NADPH oxidase and the caspase inhibitor-resistant neurotoxicity by N141I-PS2 through xanthine oxidase. Pertussis toxin greatly suppressed both toxic mechanisms by N141I-PS2, and only Galpha(o), a neuron-enriched pertussis toxin-sensitive G protein, was involved in both mechanisms. We therefore conclude that N141I-PS2 is capable of triggering multiple neurotoxic mechanisms, which can be inhibited by the combination of clinically usable inhibitors of NADPH oxidase and xanthine oxidase. This study thus provides a novel insight into the therapeutic intervention of PS2 mutant-associated FAD. PMID- 11861777 TI - Functional involvement of rat organic anion transporter 3 (rOat3; Slc22a8) in the renal uptake of organic anions. AB - Our previous kinetic analyses have shown that the transporter responsible for the renal uptake of pravastatin, an HMG-CoA reductase inhibitor, differs from that involved in its hepatic uptake. Although organic anion transporting polypeptides are now known to be responsible for the hepatic uptake of pravastatin, the renal uptake mechanism has not been clarified yet. In the present study, the involvement of rat organic anion transporter 3 (rOat3; Slc22a8) in the renal uptake of pravastatin was investigated. Immunohistochemical staining indicates the basolateral localization of rOat3 in the kidney. rOat1- and rOat3-expressed LLC-PK1 cells exhibited specific uptake of p-aminohippurate (PAH) and pravastatin, respectively, with the Michaelis-Menten constants (Km values) of 60 microM for rOat1-mediated PAH uptake and 13 microM for rOat3-mediated pravastatin uptake. Saturable uptake of PAH and pravastatin was observed in kidney slices with Km values of 69 and 11 microM, respectively. The difference in the potency of PAH and pravastatin in inhibiting uptake by kidney slices suggests that different transporters are responsible for their renal uptake. This was also supported by the difference in the degree of inhibition by benzylpenicillin, a relatively selective inhibitor of rOat3, for the uptake of PAH and pravastatin by kidney slices. These results suggest that rOat1 and rOat3 are mainly responsible for the renal uptake of PAH and pravastatin, respectively. PMID- 11861778 TI - Up-regulation of cyclooxygenase-2 by inhibition of cyclooxygenase-1: a key to nonsteroidal anti-inflammatory drug-induced intestinal damage. AB - Nonsteroidal anti-inflammatory drugs (NSAIDs) induce gastrointestinal ulceration as the adverse reaction. This effect of NSAIDs is attributable to endogenous prostaglandin (PG) deficiency caused by inhibition of cyclooxygenase (COX), yet the relation between COX inhibition and the gastrointestinal ulcerogenic property of NSAIDs remains controversial. Using selective COX inhibitors, we examined whether inhibition of COX-1 or COX-2 alone is sufficient for induction of intestinal damage in rats. Various COX inhibitors were administered p.o. in rats, and the animals were killed 24 h later. Mucosal PGE2 levels were determined by enzyme immunoassay, whereas the gene expression of COX isozymes was examined by reverse transcription-polymerase chain reaction. Nonselective COX inhibitors such as indomethacin inhibited PGE2 production and caused damage in the small intestine. Selective COX-2 inhibitors (rofecoxib or celecoxib) had no effect on the generation of PG, resulting in no damage. A selective COX-1 inhibitor (SC 560) did not cause damage, despite reducing PGE2 content. However, the combined administration of COX-1 and COX-2 inhibitors provoked intestinal damage with an incidence of 100%. COX-2 was up-regulated in the small intestine after administration of SC-560, and the PGE2 content was restored 6 h later, in a rofecoxib-dependent manner. The intestinal lesions induced by SC-560 plus rofecoxib were significantly prevented by later administration of 16,16-dimethyl PGE2. These results suggest that the intestinal ulcerogenic property of NSAID is not accounted for solely by inhibition of COX-1 and requires inhibition of COX-2 as well. The inhibition of COX-1 up-regulates COX-2 expression, and this may be a key to NSAID-induced intestinal damage. PMID- 11861779 TI - Nitric oxide synthase expression in hypertension induced by inhibition of glutathione synthase. AB - Induction of chronic oxidative stress by glutathione (GSH) depletion has been shown to cause hypertension in normal rats. This was accompanied by and perhaps in part due to inactivation and sequestration of NO by reactive oxygen species (ROS), leading to diminished NO bioavailability. This study was designed to examine renal histology, nitric oxide synthase (NOS) isotype expression, and nitrotyrosine distribution in this model. Sprague-Dawley rats were subjected to oxidative stress by administration of the GSH synthase inhibitor buthionine sulfoximine (BSO; 30 mM/l in drinking water) for 2 weeks. The controls were given tap water. Blood pressure, renal histology, tissue expression of endothelial and inducible NOS (eNOS and iNOS) and nitrotyrosine, tissue GSH content, and urinary excretion of NO metabolites (NOx) were examined. The BSO-treated group showed a 3 fold decrease in tissue GSH content, a marked elevation in blood pressure, and a significant reduction in the urinary excretion of NOx. Histological examination of kidneys revealed no significant abnormalities in either group. In addition, no significant differences were observed in either intensities or localizations of eNOS and iNOS in the kidney. However, the BSO-treated group exhibited intense accumulation in the renal tissue of nitrotyrosine, which is the footprint of NO oxidation by ROS. These observations suggest that oxidative stress-induced hypertension is not caused by either structural abnormality of or depressed NOS expression by the kidney in this model. Instead, it is associated with and perhaps partially related to enhanced renal NO inactivation by ROS and diminished NO bioavailability. PMID- 11861780 TI - Thalidomide modulates nuclear redox status and preferentially depletes glutathione in rabbit limb versus rat limb. AB - Thalidomide produces numerous birth defects, the most notable being phocomelia. Mechanisms behind thalidomide-induced malformations have not been fully elucidated, although recent evidence suggests a role for reactive oxygen species. A thalidomide-resistant (rat) and -sensitive (rabbit) species were used to compare potential inherent differences related to oxidative stress that may provide a more definitive understanding of mechanisms of thalidomide embryopathy. Limb bud cells (LBCs) were removed from the rat and rabbit embryo, dissociated, and plated in culture for 24 h. A fluorescence (6-carboxy-2',7' dichlorofluorescin diacetate; DCF) assay for oxidative stress was used with varying concentrations of thalidomide (5-100 microM). Thalidomide (100 microM) showed a 6-fold greater production of oxidative stress in rabbit cultures than in rat. Lower concentrations (50 and 25 microM) also showed a significant increase in reactive oxygen species. Confocal microscopy revealed DCF fluorescence preferentially in rabbit LBC nuclei compared with the uniform distribution of DCF fluorescence in rat LBC. Localization of glutathione (GSH) was determined using 5 chloromethylfluorescein diacetate fluorescent confocal microscopy. In rat cultures, significant thalidomide-induced GSH depletion was detected in the cytosol but the nuclei maintained its GSH content, but rabbit LBC showed significant GSH depletion in both compartments. GSH depletion was confirmed by high-performance liquid chromatography analysis. These observations provide evidence that thalidomide preferentially produces oxidative stress in the thalidomide-sensitive species but not the thalidomide-resistant species. Nuclear GSH content in the rabbit LBC is selectively modified and indicates a shift in the nuclear redox environment. Redox shifts in the nucleus may result in the misregulation of transcription factor/DNA interactions and cause defective growth and development. PMID- 11861781 TI - Platelet-endothelial cell adhesion molecule-1-directed immunotargeting to cardiopulmonary vasculature. AB - Therapeutic molecules conjugated with antibodies to the platelet-endothelial cell adhesion molecule-1 (PECAM-1) accumulate in the pulmonary endothelium after i.v. injection in mice. In this study, we characterized PECAM-directed targeting to the lung and heart after local versus systemic intravascular administration in a large animal model, pigs. Radiolabel tracing showed that 1 h post-i.v. injection, 35% of anti-PECAM versus 2.5% of control IgG had accumulated in the lungs. Infusion of anti-PECAM via a catheter placed in the right pulmonary artery (RPA) resulted in a 3-fold elevation of the uptake in the right lower lobe and 2-fold reduction of uptake in the left lobes in the lung. Cardiac uptake of anti-PECAM was negligible after i.v. and RPA infusion. In contrast, delivery with a catheter placed in the right coronary artery (RCA) resulted in a 4-fold elevation of cardiac uptake of anti-PECAM, but not IgG, compared with i.v. injection. To estimate the targeting of an active reporter enzyme, streptavidin-conjugated beta galactosidase (beta-Gal) was coupled to anti-PECAM or IgG (anti-PECAM/beta-Gal and IgG/beta-Gal) and injected into the RCA. Beta-Gal activity was markedly elevated in the heart and lungs (5- and 25-fold increased, respectively) after injection of anti-PECAM/beta-Gal, but not IgG/beta-Gal. Image analysis confirmed endothelial targeting of anti-PECAM/beta-Gal in the heart and lung. In summary, anti-PECAM antibody conjugates deliver agents to the pulmonary endothelium regardless of injection route, whereas local arterial infusion permits targeting to the cardiac vasculature. This paradigm may be useful for drug targeting to endothelium in lungs, heart, and possibly other organs. PMID- 11861782 TI - Neuroprotective effect of cilostazol against focal cerebral ischemia via antiapoptotic action in rats. AB - This study examined the protective effects of cilostazol on cerebral infarcts produced by subjecting rats to 2-h occlusion of the left middle cerebral artery followed by 24-h reperfusion. The ischemic cerebral infarct consistently involved the cortex and striatum. The infarct size was significantly reduced, when rats received 10 mg/kg cilostazol intravenously 5 min or 1 h after the completion of 2 h ischemia. Cyclic AMP level was significantly elevated in the cortex of 4- and 12-h reperfusion (P < 0.01) following treatment with cilostazol (10 mg/kg, 5 min after 2-h ischemia) accompanied by decreased tumor necrosis factor-alpha level. Samples from the regions corresponding to the penumbra showed markedly reduced Bcl-2 protein level and, in contrast, high levels of Bax protein and cytochrome c release. Cilostazol decreased Bax protein and cytochrome c release and increased the levels of Bcl-2 protein. Cilostazol (10(-7)-10(-5) M) potently and concentration dependently scavenged hydroxyl and peroxyl radicals. In conclusion, cilostazol treatment decreases ischemic brain infarction in association with inhibition of apoptotic and oxidative cell death. PMID- 11861783 TI - Bucindolol exerts agonistic activity on the propranolol-insensitive state of beta1-adrenoceptors in human myocardium. AB - In congestive heart failure patients, treatment with beta-adrenoceptor antagonists improves symptoms and decreases mortality. However, intrinsic sympathomimetic activity of beta-adrenoceptor antagonists might be disadvantageous in chronic heart failure. The nonselective beta1- and beta2 adrenoceptor antagonist bucindolol has failed to decrease mortality in clinical trials. A putative beta4-adrenoceptor, which mediates positive inotropic effects by activation of the adenylate cyclase has been described. Recently, this putative beta4-adrenoceptor has been identified to be a propranolol-insensitive state of the beta1-adrenoceptor. The present study aimed to characterize whether bucindolol exhibits agonistic activity on this atypical beta1-adrenoceptor state as one possible reason for clinical inefficiency. For comparison (S)-4-(3'-t butylamino-1'-hydroxypropoxy)-benzimidozole-2 (CGP 12177), metoprolol, and nebivolol were investigated. Bucindolol did not reveal intrinsic sympathomimetic activity in electrically driven (1 Hz, 37 degrees C), forskolin-stimulated, left ventricular papillary muscle strips (donor hearts, nonfailing; n = 5) and in right auricular trabeculae (bypass operation; n = 4). Functional studies on the propranolol-insensitive state of beta1-adrenoceptors were performed in isolated muscle preparations after beta1- and beta2-adrenoceptor antagonism (propranolol, 1 microM), inhibition of beta3-mediated inotropic effects (N-nitro-L-arginine, 100 microM) and forskolin treatment (0.3 microM). Positive inotropic response to stimulation of atypical state beta1-adrenoceptors could be demonstrated in right auricular as well as left ventricular human myocardium (CGP 12177 treatment, 10 microM). Under these conditions, also bucindolol, but not metoprolol and nebivolol, significantly increased contractility (all 10 microM). In conclusion, bucindolol but not metoprolol or nebivolol mediate positive inotropic effects in human myocardium due to activation of atypical state beta1-adrenoceptors. Thus, the agonistic activity of bucindolol may influence outcome in heart failure patients. PMID- 11861784 TI - Effects of 3-phenyl-4-[[4-[2-(1-piperidinyl)ethoxy]phenyl]methyl]- 2H-1 benzopyran-7-ol (CHF 4056), a novel nonsteroidal estrogen agonist/antagonist, on reproductive and nonreproductive tissue. AB - We have discovered a new, nonsteroidal, estrogen agonist/antagonist, 3-phenyl-4 [[4-[2-(1-piperidinyl)ethoxy]phenyl] methyl]-2H-1-benzopyran-7-ol (CHF 4056). The aim of this study was to determine the effects of CHF 4056 on a series of parameters (body weight, uteri, serum cholesterol, and bones) that were previously shown to be sensitive to estrogens and to selective estrogen receptor modulators (SERMs). CHF 4056 is a benzopyran derivative that binds with high affinity to the human estrogen receptors alpha and beta (dissociation constant K(i) of 0.041 and 0.157 nM, respectively). In immature rats, CHF 4056 induced a full estrogen antagonism (half-maximal efficacious dose = 0.33 mg/kg x day p.o.) coupled with a lack of uterine stimulatory activity, whereas the structurally related SERM levormeloxifene demonstrated a maximal partial agonist effect of approximately 65% that of 17alpha-ethynyl estradiol (EE2). In ovariectomized (OVX) rats, CHF 4056 (0.1-1 mg/kg x day p.o. for 4 weeks) significantly reduced OVX-induced bone loss in the lumbar spine L1-4 and OVX-induced increase in serum osteocalcin. These protective effects on bone tissue were comparable with those of 0.1 mg/kg x day EE2. In the same experimental conditions, serum cholesterol was significantly lower in the CHF 4056-treated animals, compared with vehicle treated OVX rats. In line with the results observed in immature rats, also in OVX rats CHF 4056 diverged dramatically from EE2 and levormeloxifene in its lack of significant estrogenic effects on uterine tissue. In conclusion, CHF 4056 is a new SERM that produces beneficial effects on bone and cholesterol levels, while maintaining antagonist effects on the uterus. PMID- 11861785 TI - Homologous mutations near the junction of the sixth transmembrane domain and the third extracellular loop lead to constitutive activity and enhanced agonist affinity at all muscarinic receptor subtypes. AB - Previous studies have found that a mutation near the junction of the sixth transmembrane domain (TM6) and the third extracellular loop of the M5 muscarinic receptor leads to constitutive activation and enhanced agonist affinity for the mutated receptor. These results were consistent with the extended ternary complex model, which predicts a correlation between agonist affinity and constitutive activity. We have introduced the homologous mutation into all five subtypes of the highly conserved muscarinic receptor family; SerThr-->TyrPro was introduced into M1 and M5, and AsnThr-->TyrPro was introduced into M2, M3, and M4. In binding assays, these mutations produced increases in affinities toward acetylcholine and carbachol that ranged from 5-fold at the M2 receptor to 15- to 20-fold at M1, M3, and M4, to 40-fold at M5. In functional assays, all five mutant receptors exhibited constitutive activity, at levels ranging between 30 and 80% of the maximal response elicited by carbachol. In every case, the muscarinic antagonist atropine inhibited this constitutive activity with high affinity. Thus, despite differences in effector coupling and in wild-type sequence at the mutation site, all five subtypes were activated by this mutation at the top of TM6. Previous studies of the M5 subtype have indicated that TM6 is a ligand-dependent switch that sets the activation state of the receptor. Based on the results of the present study, it is possible that TM6 represents a general switch for the activation of the muscarinic receptor family. PMID- 11861786 TI - Inorganic lead activates the mitogen-activated protein kinase kinase-mitogen activated protein kinase-p90(RSK) signaling pathway in human astrocytoma cells via a protein kinase C-dependent mechanism. AB - We have previously reported that lead acetate activates protein kinase Calpha (PKCalpha) and induces DNA synthesis in human 1321N1 astrocytoma cells. In this study, we investigated the ability of lead to activate the mitogen-activated protein kinase (MAPK) cascade. We found that exposure to lead acetate (1-50 microM) resulted in concentration- and time-dependent activation of MAPK (extracellular signal responsive kinase 1/2), as shown by increased phosphorylation and increased kinase activity. This effect was significantly reduced by the PKC-specific inhibitor bisindolylmaleimide (GF109203X), by down regulation of PKC with 12-O-tetradecanoyl-phorbol 13-acetate, by a pseudosubstrate to PKCalpha, and by selective down-regulation of PKCalpha by prior lead exposure. Lead was also shown to activate MAPK kinase (MEK1/2), and this effect was mediated by PKC. Two MEK inhibitors, 2-(2'-amino-3' methoxyphenol)-oxanaphthalen-4-one (PD98059) and 1,4-diamino-2,3-dicyano-1,4 bis[2-aminophenylthio]butadiene (UO126), blocked lead-induced MAPK activation and inhibited lead-induced DNA synthesis, as measured by incorporation of [methyl 3H]thymidine into cell DNA. The 90 kDa ribosomal S6 protein kinase, p90(RSK), a substrate of MAPK, was also found to be activated by lead in a PKC- and MAPK dependent manner. Stimulation of DNA synthesis by lead in astrocytoma cells may be of interest in light of the observed association between exposure to lead and an increased risk of astrocytomas. PMID- 11861787 TI - Enhanced induction of cytochrome P450 enzymes and CAR binding in TNF (p55(-/ )/p75(-/-)) double receptor knockout mice following phenobarbital treatment. AB - Phenobarbital (PB) is a well characterized inducer of cytochrome P450 (P450) 2B and 3A subfamilies. Several proinflammatory cytokines have been shown to negatively modulate the induction of P450 by PB. In addition, PB is known to elicit an inflammatory mitogenic effect on the liver. To date, no studies have evaluated the PB induction profile of hepatic P450 in the absence of an intact tumor necrosis factor-alpha (TNFalpha) response. To test the hypothesis that endogenous TNFalpha signaling modulates hepatic P450 induction by PB in vivo, PB induction was examined in TNF (p55(-/-)/p75(-/-)) double receptor knockout mice (ko-TNF) and wild-type mice (wt-TNF). CYP2B- and CYP3A-associated activities and protein content were induced to a significantly greater extent (p < 0.05) in ko TNF mice compared with wt-TNF mice. In parallel with enhanced CYP2B induction, an apparent elevation in the nuclear accumulation of the principal regulatory protein for transcription of CYP2B genes, the constitutively activated receptor (CAR), was detected in ko-TNF nuclear extracts following PB treatment. Additionally, nuclear factor kappa-B binding was induced by PB in wt-TNF mice, but not in ko-TNF mice, indicating that the hepatic inflammatory response following PB treatment differed between wt-TNF and ko-TNF mice. These data demonstrate that endogenous TNFalpha signaling modulates PB induction of hepatic CYP2B and CYP3A isoforms in vivo. Further, the data presented herein suggest that endogenous TNFalpha signaling influences PB induction of CYP2B through inhibition of CAR nuclear accumulation. PMID- 11861788 TI - Serotonergic attenuation of the reinforcing and neurochemical effects of cocaine in squirrel monkeys. AB - Preclinical studies have documented that serotonin (5-HT) can modulate the behavioral effects of cocaine. The present study examined the ability of 5-HT to attenuate the reinforcing and neurochemical effects of cocaine in nonhuman primates. In squirrel monkeys trained to self-administer cocaine (0.1 and 0.3 mg/injection) under a second-order schedule of i.v. drug delivery, the 5-HT uptake inhibitor alaproclate (3.0 and 10.0 mg/kg) and the 5-HT direct agonist quipazine (0.3-1.0 mg/kg) decreased response rates at doses that had no significant effect on behavior maintained by an identical schedule of stimulus termination. The neurochemical bases of the observed drug interactions on behavior were investigated further using in vivo microdialysis techniques in a separate group of awake monkeys to monitor drug-induced changes in extracellular dopamine (DA). Cocaine (1.0 mg/kg) elevated the concentration of DA in the caudate nucleus to approximately 300% of basal levels. Pretreatment with alaproclate or quipazine attenuated cocaine-induced increases in extracellular DA at the same pretreatment doses that decreased cocaine self-administration. The results obtained suggest that increasing brain 5-HT activity can attenuate the reinforcing effects of cocaine, ostensibly by decreasing the ability of cocaine to elevate extracellular DA in brain areas that mediate the behavioral effects. These findings extend those reported previously for the behavioral-stimulant effects of cocaine and identify a potential neurochemical mechanism underlying drug interactions on behavior. PMID- 11861789 TI - NADPH-dependent metabolism of estrone by human liver microsomes. AB - We characterized the NADPH-dependent metabolism of estrone (E1) by liver microsomes of 21 male and 12 female human subjects. The structures of 11 hydroxylated or keto metabolites of E1 formed by human liver microsomes were identified by chromatographic and mass spectrometric analyses. 2-Hydroxylation of E1 was the dominant metabolic pathway with all human liver microsomes tested. E1 is more prone to form catechol estrogens (particularly 4-OH-E1) than 17beta estradiol (E2) and the average ratio of E1 4-hydroxylation to 2-hydroxylation (0.24) was slightly higher than the ratio of E2 4- to 2-hydroxylation (0.20, P < 0.001). An unidentified monohydroxylated E1 metabolite (y-OH-E1) was found to be one of the major metabolites formed by human liver microsomes of both genders. 6beta-OH-E1, 16alpha-OH-E1, and 16beta-OH-E1 were also formed in significant quantities. 16alpha-hydroxylation was not a major pathway for E1 metabolism. The overall profiles for the E1 metabolites formed by male and female human liver microsomes were similar, and their average rates were not significantly different. Hepatic CYP3A4/5 activity in both male and female liver microsomes correlated strongly with the rates of formation of several hydroxyestrogen metabolites. The dominant role of hepatic CYP3A4 and CYP3A5 in the formation of these hydroxyestrogen metabolites was further confirmed by incubations of human CYP3A4 or CYP3A5 with [3H]E1 and NADPH. Notably, human CYP3A5 has very high relative activity for E1 4-hydroxylation, exceeding its activity for E1 2 hydroxylation by approximately 100%. It will be of interest to determine the potential biological functions associated with any of the E1 metabolites identified in our present study. PMID- 11861790 TI - Effects of individual and concurrent stimulation of striatal D1 and D2 dopamine receptors on electrophysiological and behavioral output from rat basal ganglia. AB - Bilateral infusions of d-amphetamine into the rat ventral-lateral striatum (VLS) were previously shown to cause a robust behavioral activation that was correlated temporally with a net increase in firing of substantia nigra pars reticulata (SNpr) neurons, a response opposite predictions of the basal ganglia model. The current studies assessed the individual and cooperative contributions of striatal D1 and D2 dopamine receptors to these responses. Bilateral infusions into VLS of the D1/D2 agonist apomorphine (10 microg/microl/side) caused intense oral movements and sniffing, and an overall increase in SNpr cell firing to 133% of basal rates, similar to effects of d-amphetamine. However, when striatal D2 receptors were stimulated selectively by infusions of quinpirole (30 microg/microl/side) + the D1 antagonist R-(+)-7-chloro-8-hydroxy-3-methyl-1 phenyl-2,3,4,5-tetrahydro-1H-3-benzazepine (SCH 23390; 10 microg/microl/side), no behavioral response and only modest and variable changes in SNpr cell firing were observed. Selective stimulation of striatal D1 receptors by (+/-) 6-chloro-APB hydrobromide (SKF 82958; 10 microg/microl/side) + the D2 antagonist cis-N-(1 benzyl-2-methyl-pyrrolidin-3-yl)-5-chloro-2-methoxy-4-methyl-aminobenzamide (YM 09151-2; 2 microg/microl/side) caused a weak but sustained increase in oral movements and modestly increased SNpr cell firing, but neither response was of the magnitude observed with apomorphine. When the two agonists were infused concurrently, however, robust oral movements and sniffing again occurred over the same time period that a majority of SNpr cells exhibited marked, sometimes extreme and fluctuating, changes in firing (net increase, 117% of basal rates). These data confirm that concurrent striatal D1/D2 receptor stimulation elicits a strong motor activation that is correlated temporally with a net excitation rather than inhibition of SNpr firing, and reveal that D1 and D2 receptors interact synergistically within the striatum to stimulate both forms of output. PMID- 11861791 TI - Activation of poly(ADP-ribose) polymerase contributes to development of doxorubicin-induced heart failure. AB - Activation of the nuclear enzyme poly(ADP-ribose) polymerase (PARP) by oxidant mediated DNA damage is an important pathway of cell dysfunction and tissue injury in conditions associated with oxidative stress. Increased oxidative stress is a major factor implicated in the cardiotoxicity of doxorubicin (DOX), a widely used antitumor anthracycline antibiotic. Thus, we hypothesized that the activation of PARP may contribute to the DOX-induced cardiotoxicity. Using a dual approach of PARP-1 suppression, by genetic deletion or pharmacological inhibition with the phenanthridinone PARP inhibitor PJ34, we now demonstrate the role of PARP in the development of cardiac dysfunction induced by DOX. PARP-1+/+ and PARP-1-/- mice received a single injection of DOX (25 mg/kg i.p). Five days after DOX administration, left ventricular performance was significantly depressed in PARP 1+/+ mice, but only to a smaller extent in PARP-1-/- ones. Similar experiments were conducted in BALB/c mice treated with PJ34 or vehicle. Treatment with a PJ34 significantly improved cardiac dysfunction and increased the survival of the animals. In addition PJ34 significantly reduced the DOX-induced increase in the serum lactate dehydrogenase and creatine kinase activities but not metalloproteinase activation in the heart. Thus, PARP activation contributes to the cardiotoxicity of DOX. PARP inhibitors may exert protective effects against the development of severe cardiac complications associated with the DOX treatment. PMID- 11861792 TI - Leukotriene B4 production in human mononuclear phagocytes is modulated by interleukin-4-induced 15-lipoxygenase. AB - The aim of this study was to evaluate the consequences of interleukin (IL)-4 induced 15-lipoxygenase (15-LO) expression on leukotriene B4 (LTB4) synthesis in human monocytes. Human monocytes incubated for 24, 48, and 72 h with IL-4 (10 ng/ml) were stimulated with Ca2+-ionophore A23187 (calcimycin; 5 microM) or opsonized zymosan. 15(S)-hydroxyeicosatetraenoic acid [15(S)-HETE], LTB4, and arachidonic acid (AA) release were measured by high-performance liquid chromatography/radioimmunoassay, liquid chromatography/tandem mass spectrometry (LC/MS/MS), or gas chromatography/mass spectrometry. 15-LO activity was evaluated in AA-treated monocytes. 15-LO, 5-lipoxygenase (5-LO) and 5-LO activating protein (FLAP) expression were analyzed by reverse transcription-polymerase chain reaction. Neutrophil chemotactic activity was evaluated using a microtaxis chamber assay. A23187-induced synthesis of 15(S)-HETE was significantly increased after treatment with IL-4 (10 ng/ml) for 48 and 72 h (p < 0.001). Concomitant decrease of LTB4 release was observed after 72 h of incubation with IL-4 (p < 0.001). LC/MS/MS analysis confirmed the production of 15(S)-HETE and the significant inhibition of LTB4 synthesis in IL-4-treated monocyte after challenge with opsonized zymosan. IL-4 treatment induced 15-LO enzymatic activity as well as 15-LO mRNA, but did not affect either 5-LO or FLAP mRNA expression in monocytes. Supernatant from IL-4-treated monocytes showed significantly lower neutrophil chemotactic activity than controls. 15(S)-HETE significantly inhibited LTB4 production induced by A23187-stimulated human monocytes without affecting AA release. IL-4-induced expression of 15-LO in monocytes caused a significant reduction of LTB4 production. Whereas this effect did not reflect changes in 5-LO and FLAP mRNA expression, synthetic 15(S)-HETE was able to significantly inhibit the synthesis of LTB4, without affecting AA release. PMID- 11861793 TI - Nicotine reduces the incidence of type I diabetes in mice. AB - Nicotine has been previously shown to have immunosuppressive actions. Type I diabetes is an autoimmune disease resulting from the specific destruction of the insulin-producing pancreatic beta-cells. Thus, we hypothesized that nicotine may exert protective effects against type I diabetes. The multiple low-dose streptozotocin (MLDS)-induced model and spontaneous nonobese diabetic (NOD) mouse model of type I diabetes were used to assess whether nicotine could prevent this autoimmune disease. Blood glucose levels, diabetes incidence, pancreas insulin content, and cytokine levels were measured in both models of diabetes, both to asses the level of protection exerted by nicotine and to further investigate its mechanism of action. Nicotine treatment reduced the hyperglycemia and incidence of disease in both the MLDS and NOD mouse models of diabetes. Nicotine also protected against the diabetes-induced decrease in pancreatic insulin content observed in both animal models. The pancreatic levels of the Th1 cytokines interleukin (IL)-12, IL-1, tumor necrosis factor (TNF)-alpha, and interferon (IFN)-gamma were increased in both MLDS-induced and spontaneous NOD diabetes, an effect prevented by nicotine treatment. Nicotine treatment increased the pancreatic levels of the Th2 cytokines IL-4 and IL-10. Nicotine treatment reduces the incidence of type I diabetes in two animal models by changing the profile of pancreatic cytokine expression from Th1 to Th2. PMID- 11861794 TI - Reversal of ethanol-seeking behavior by D1 and D2 antagonists in an animal model of relapse: differences in antagonist potency in previously ethanol-dependent versus nondependent rats. AB - Mesocorticolimbic dopamine (DA) transmission has been implicated in the consummatory and, more recently, the incentive-motivational aspect of ethanol's actions. The purpose of this study was to test whether ethanol-seeking behavior induced by an ethanol-associated contextual stimulus is sensitive to antagonism of DA transmission. Male Wistar rats were trained to orally self-administer 10% ethanol and to associate olfactory discriminative stimuli with the availability of ethanol (S(+)) versus nonreward (S(-)). Ethanol-reinforced operant responding then was extinguished by withholding ethanol and the associated S(+). After reaching a predetermined extinction criterion, reinstatement tests were conducted in which the animals were presented noncontingently with only the S(+) or S(-). Exposure to the S(+) but not the S(-) reinstated responding at the previously active lever. The D1 antagonist R(+)-7-chloro-8-hydroxy-3-methyl-1-phenyl-2,3,4,5 tetrahydro-1H-3-benzazepine hydrochloride (SCH23390; 5, 10, 50 microg/kg s.c.) and the D2 antagonist eticlopride (5, 10, 50 microg/kg s.c.) dose dependently decreased the number of S(+)-induced responses and increased response latency. During a second test, conducted in the same rats, 3 weeks after withdrawal from a 12-day ethanol vapor inhalation procedure, the response-reinstating efficacy of the S(+) remained unaltered. However, the potency of both DA antagonists to inhibit the S(+)-induced drug-seeking response was significantly increased. The results confirm that ethanol-related contextual stimuli reliably elicit drug seeking behavior and suggest that this effect requires activation of DA neurotransmission. The results also indicate that chronic ethanol exposure produces changes in D1 and D2 receptor function that lead to enhanced sensitivity to the behavioral effects of antagonists for these receptors. PMID- 11861795 TI - Differential modulation of signaling pathways and apoptosis of ras-transformed 10T1/2 cells by the depsipeptide FR901228. AB - (E)-(1S,4S,10S,21R)-7-[(Z)-ethylidene]-4,21-diisopropyl-2-oxa-12,13-dithia 5,8,20,23-tetraazabicyclo[8,7,6]-tricos-16-ene-3,6,9,19,22-pentanone (FR901228), a natural anticancer depsipeptide, induces apoptosis of ras-transformed 10T1/2 cells whereas it induces growth arrest of nontransformed counterpart cells in G0/G1 phase of the cell cycle. Our study of the effect of FR901228 treatment on intracellular signaling pathways reveals a discriminating activity of FR901228 to regulate signaling cascades differently in ras-transformed 10T1/2 cells and nontransformed counterpart cells. Induction of apoptosis of ras-transformed cells by FR901228 correlates with suppression of the extracellular signal-regulated kinase (ERK) signaling pathway through reduction of Raf expression and deactivation of Mek and Erk, inhibition of the phosphoinositide-3 kinase (PI3-K) pathway indexed by suppression of Akt activity, suppression of p38 activity, and activation of caspase-3. Expression of p21(Cip1) is not induced in ras transformed cultures undergoing apoptosis induced by FR901228. In contrast, FR901228 induces p21(Cip1) expression in nontransformed counterpart cultures growth-arrested in G0/G1 that is also accompanied by moderate induction of the kinase activities of Raf, Mek, Erk, and Akt, but not accompanied by activation of caspase-3 or changes in p38 activity. Our study indicates a potential value of FR901228 in the treatment of cancer cells involving aberrant regulation of Ras through preferential induction of the caspase cascade and suppression of the ERK, PI3-K, and p38 pathways. PMID- 11861796 TI - Predominance of delta-opioid-binding sites in the porcine enteric nervous system. AB - The antidiarrheal and constipating actions of opioids are mediated in part by enteric neurons, which lie within the wall of the small intestine and colon, but the differential expression of specific, high-affinity opioid-binding sites in ganglionated plexuses within functionally distinct intestinal segments has not been examined. We determined the saturation binding characteristics under Na+ free conditions of the nonselective opioid receptor (OPR) ligand [3H][(5alpha,7alpha)-17-(cyclopropylmethyl)-4,5-epoxy-18,19-dihydro-3-hydroxy-6 methoxy-alpha, alpha-dimethyl-6,14-ethenomorphinan-7-methanol] (diprenorphine) and the respective delta-, kappa-, and mu-OPR ligands [3H]naltrindole, D (5alpha,7alpha,8beta)-(-)-N-methyl-N-[7-(1-pyrrolidinyl)-1-oxoaspiro-(4,5)dec-8 yl]benzeneacetamide ([3H]U-69,593), and [3H][D-Ala2,N-Me-Phe4,Gly5-ol]-enkephalin (DAMGO) in neuronal membranes isolated from myenteric and submucosal plexuses of porcine small intestine and colon. Naloxone-displaceable [3H]diprenorphine binding sites (KD values ranging from 0.2-0.5 nM and Bmax = 50-95 fmol/mg of protein) were found in both subregions from all gut segments examined. High affinity [3H]naltrindole sites (KD = 60-140 pmol) were at highest densities (approximately 60 fmol/mg of protein) in submucosal plexus of the ileum and distal colon myenteric plexus and were at lowest densities (8-9 fmol/mg of protein) in the submucosal plexuses of cecum and distal colon. [3H]U-69,593 sites (KD = 0.3-4 nM) were present only in the myenteric plexuses of all segments examined, with highest densities in cecum and proximal colon (44-47 fmol/mg of protein). [3H]DAMGO-binding sites were expressed at relatively low densities in the enteric plexuses of all gut regions. These results indicate that delta-OPRs predominate in the porcine enteric nervous system with a more circumscribed expression of kappa- and mu-OPRs. PMID- 11861797 TI - Functional characterization of adenosine receptors and coupling to ATP-sensitive K+ channels in Guinea pig urinary bladder smooth muscle. AB - Although multiple adenosine receptors have been identified, the subtype and underlying mechanisms involved in the relaxation response to adenosine in the urinary bladder remain unclear. The present study investigates changes in the membrane potential, as assessed by fluorescence-based techniques, of bladder smooth muscle cells by adenosine receptor agonists acting via ATP-sensitive potassium (K(ATP)) channels. Membrane hyperpolarization evoked by adenosine and various adenosine receptor subtype-selective agonists was attenuated or reversed by the K(ATP) channel blocker glyburide. Comparison of adenosine receptor agonist potencies eliciting membrane potential effects showed a rank order of potency 5' N-ethyl-carboxamido adenosine (NECA; -log EC50 = 7.97) approximately 2-p-(2 carboxethyl)phenethyl-amino-5'-N-ethylcarboxamidoadenosine hydrochloride (CGS 21680; 7.65) > 2-chloro adenosine (5.90) approximately 2-chloro-N6 cyclopentyladenosine (CCPA; 5.51) approximately N6-cyclopentyladenosine approximately N6-(R)-phenylisopropyladenosine > 2-chloro- N6-(3-iodobenzyl) adenosine-5'-N-methyl-carboxamide (2Cl-IBMECA; 4.78). Membrane potential responses were mimicked by forskolin, a known activator of adenylate cyclase, and papaverine, a phosphodiesterase inhibitor. The A(2A)-selective antagonist 4-(2-[7 amino-2-(2-furyl)[1,2,4]triazolo[2,3-a][1,3,5]triazin-5-yl-amino] ethyl)phenol (ZM-241385), and the adenylate cyclase inhibitor N-(cis-2-phenyl-cyclopentyl) azacyclotridecan-2-imine-hydrochloride (MDL-12330A) inhibited the observed change in membrane potential evoked by adenosine and adenosine-receptor agonists. The rank order potency for relaxation of K+-stimulated guinea pig bladder strips, NECA (-log EC50 = 6.41) approximately CGS-21680 (6.38) > 2-chloro adenosine (5.90) >> CCPA approximately 2Cl-IBMECA (>4.0) was comparable to that obtained from membrane potential measurements. Collectively, these studies demonstrate that adenosine-evoked membrane hyperpolarization and relaxation of bladder smooth muscle is mediated by A(2A) receptor-mediated activation of K(ATP) channels via adenylate cyclase and elevation of cAMP. PMID- 11861799 TI - Effects of tachykinin NK1 receptor antagonists on the viscerosensory response caused by colorectal distention in rabbits. AB - Irritable bowel syndrome (IBS) is a common disorder mainly characterized by altered bowel habits and visceral pain. In this study, we investigated the role of tachykinin NK1 receptors in the visceral pain response (abdominal muscle contraction) caused by colorectal distention in rabbits previously subjected to colonic irritation, using the selective tachykinin NK1 receptor antagonists TAK 637 [(aR,9R)-7-[3,5-Bis(trifluoromethyl)benzyl]-8,9,10,11-tetrahydro-9-methyl-5 (4-methylphenyl)-7H-[1,4] diazocino[2,1-g][1,7]naphthyridine-6,13-dione] and (+/ )-CP-99,994 (+/-)-(2S,3S)-3-(2-methoxybenzylamino)-2-phenylpiperidine. Intracolorectal administration of 0.8% acetic acid solution enhanced the nociceptive response to colorectal distention, producing a significant increase in the number of abdominal muscle contractions. Under these conditions, intraduodenal TAK-637 (0.1-3 mg/kg) dose dependently decreased the number of distention-induced abdominal contractions, and a significant inhibitory effect was observed with doses of 0.3 to 3 mg/kg. Another tachykinin NK1 antagonist, (+/ )-CP-99,994, also reduced the number of abdominal contractions. In contrast, the enantiomer of TAK-637 (which has very weak tachykinin NK1 receptor antagonistic activity), trimebutine maleate, ondansetron, and atropine sulfate did not inhibit the abdominal response. The main metabolite of TAK-637, which has more potent tachykinin NK1 receptor antagonistic activity but permeates the central nervous system less well than TAK-637, produced less inhibition of the viscerosensory response. When given intrathecally, TAK-637 and (+/-)-CP-99,994 markedly reduced the number of abdominal contractions. These results suggest that tachykinin NK1 receptors play an important role in mediating visceral pain and that TAK-637 inhibits the viscerosensory response to colorectal distention by antagonizing tachykinin NK1 receptors, mainly in the spinal cord. They also suggest that TAK 637 may be useful in treating functional bowel disorders such as IBS. PMID- 11861798 TI - Human organic anion transporters and human organic cation transporters mediate renal antiviral transport. AB - Renal excretion is an important elimination pathway for antiviral agents, such as acyclovir (ACV), ganciclovir (GCV), and zidovudine (AZT). The purpose of this study was to elucidate the molecular mechanisms of renal ACV, GCV, and AZT transport using cells stably expressing human organic anion transporter 1 (hOAT1), hOAT2, hOAT3, and hOAT4, and human organic cation transporter 1 (hOCT1) and hOCT2. Time- and concentration-dependent uptake of ACV and GCV was observed in hOAT1- and hOCT1-expressing cells. In contrast, uptake of valacyclovir, L valyl ester of ACV, was observed only in hOAT3-expressing cells. On the other hand, AZT uptake was observed in hOAT1-, hOAT2-, hOAT3-, and hOAT4-expressing cells. The Km values of ACV uptake by hOAT1 and hOCT1 were 342.3 and 151.2 microM, respectively, whereas those of GCV uptake by hOAT1 and hOCT1 were 895.5 and 516.2 microM, respectively. On the other hand, the Km values of AZT uptake by hOAT1, hOAT2, hOAT3, and hOAT4 were 45.9, 26.8, 145.1, and 151.8 microM, respectively. In addition, probenecid weakly inhibited the hOAT1-mediated ACV uptake. In conclusion, these results suggest that hOAT1 and hOCT1 mediate renal ACV and GCV transport, whereas hOAT1, hOAT2, hOAT3, and hOAT4 mediate renal AZT transport. In addition, L-valyl ester appears to be important in differential substrate recognition between hOAT1 and hOAT3. hOAT1 may not be the molecule responsible for the drug interaction between ACV and probenecid. PMID- 11861800 TI - Endogenous opioids in dopaminergic cell body regions modulate amphetamine-induced increases in extracellular dopamine levels in the terminal regions. AB - Opioid antagonists attenuate behavioral effects of amphetamine and amphetamine induced increases in extracellular dopamine levels in nucleus accumbens and striatum of rats but do not alter those effects of cocaine. This study was performed to determine 1) if the effect of opioid antagonists on the dopamine response to amphetamine is mediated in either the terminal or cell body region of the nigrostriatal and mesolimbic pathways, and 2) if the enkephalinase inhibitor thiorphan, which slows degradation of endogenous opioid peptides, increases the dopamine response to amphetamine but not to cocaine. Microdialysis probes were placed either into a dopaminergic terminal region or into both a terminal and cell body region of rats. Naloxone methiodide (1.0 microM), a lipophobic opioid antagonist, was administered into either the terminal or cell body region by reverse dialysis, whereas extracellular dopamine was collected in the terminal region. Increases in extracellular dopamine in nucleus accumbens and striatum caused by amphetamine (0.1-6.4 mg/kg, s.c.) were reduced significantly (28-39%) by naloxone methiodide administered into either substantia nigra or ventral tegmentum but not into terminal regions. Thiorphan (10 microM) administered into substantia nigra increased significantly the dopamine response to amphetamine in the ipsilateral striatum by as much as 42% but did not affect the dopamine response to cocaine (3.0-56 mg/kg, i.p.). These results suggest that amphetamine promotes release of endogenous opioids, which, through actions in the ventral tegmentum and substantia nigra, contribute to amphetamine-induced increases in extracellular dopamine in the nucleus accumbens and striatum. PMID- 11861801 TI - Distinct pathways of apoptosis triggered by FTY720, etoposide, and anti-Fas antibody in human T-lymphoma cell line (Jurkat cells). AB - 2-amino-2-[2-(4-octylphenyl)ethyl] propane-1,3-diol hydrochloride (FTY720), a synthetic product derived from a metabolite of Isaria sinclairii, has been demonstrated to have a potent immunosuppressive activity that induces apoptotic cell death in T cells and several other cell lines. In this study, using the human T-lymphoma cell line, Jurkat cells, we investigated the apoptotic signal transduction mediated by FTY720, in particular comparing its role on the cleavage of caspases, with that mediated by etoposide or anti-Fas antibody. All of these agents cleaved caspases, inducing their active form in the affected cells. Pretreatment with a broad caspase inhibitor [benzyloxycarbonyl-Val-Ala-Asp-(Ome) fluoromethyl ketone] markedly decreased the incidence of apoptotic cells induced by FTY720, etoposide, and anti-Fas antibody, through the abrogation of cleavage of Bid, poly(ADP-ribose) polymerase, and caspases 3, 8, and 9. The overexpression of Bcl-2 gene prevented FTY720- and etoposide-mediated apoptosis, but not Fas mediated apoptosis. In addition, mitochondria were demonstrated to play a critical role in FTY720-triggered cell death, suggesting that this drug has a potent anticancer activity. PMID- 11861802 TI - The role of ATP-sensitive potassium channels in neutrophil migration and plasma exudation. AB - Neutrophil activation and migration during an inflammatory response is preceded or accompanied by plasma membrane electrical changes. Besides changes in calcium currents, neutrophils have a high permeability to potassium, mainly through potassium channels. However, the significance of potassium channels in neutrophil physiology is still unclear. Here, we show that the treatment of rats with the ATP-sensitive potassium channel blocker glibenclamide (4, 20, or 40 micromol/kg) dose dependently decreased carrageenan-, N-formyl-methionyl-leucyl-phenylalanine (fMLP)-, and lipopolysaccharide-induced neutrophil influx and fluid leakage into the interpleural space. On the other hand, minoxidil (an ATP-sensitive potassium channel opener; 25, 50, and 100 micromol/kg) increased both neutrophil influx and fluid leakage induced by a submaximal dose of carrageenan. In addition, in vitro human neutrophil chemotaxis induced by leukotriene B4 or fMLP (both 1 microM) was fully blocked by glibenclamide (10, 30, and 100 microM) or tetraethylammonium (a nonselective potassium channel blocker; 1, 3, and 10 mM). Thus, our results disclose the possibility that ATP-sensitive potassium channels may have a role in neutrophil migration and chemotaxis and plasma exudation in the inflammatory response. PMID- 11861803 TI - Close relationship between the platelet activation marker CD62 and the granular release of platelet-derived growth factor. AB - The expression of CD62 on the surface of platelets is considered to be an indicator of platelet degranulation and secretion. We characterized the relationship between CD62 expression and platelet-derived growth factor (PDGF)(AB) and PDGF(BB) secretion in response to thrombin-receptor activating peptide (TRAP). The principal findings were 1) expression of CD62 as a constituent of platelet alpha-granule membrane and secretion of PDGF, an important ingredient of alpha-granules, can be stimulated by TRAP-induced activation in a dose-dependent fashion; 2) the activation marker and secretion product are closely correlated with each other; and 3) changes in the CD62 expression induced by a drug, namely clopidogrel, or by a disease, namely diabetes, are paralleled by changes in PDGF secretion. Although CD62 is perceived as an activation marker of platelets indicating enhanced aggregability and secretion of alpha-granular content, the proof that the CD62 status and its modifications reflect directly the actual secretion of the most important platelet mitogen, PDGF, has so far not been given. This ex vivo-in vitro study shows that at least for the activation pathway provided by the PAR-1 receptor for which TRAP is the selective agonist, CD62 expression on platelets could be a surrogate for their secretory activity. PMID- 11861804 TI - A novel neurotrophic property of glucagon-like peptide 1: a promoter of nerve growth factor-mediated differentiation in PC12 cells. AB - The insulinotropic hormone glucagon-like peptide-1 (7-36)-amide (GLP-1) has potent effects on glucose-dependent insulin secretion, insulin gene expression, and pancreatic islet cell formation and is presently in clinical trials as a therapy for type 2 diabetes mellitus. We report on the effects of GLP-1 and two of its long-acting analogs, exendin-4 and exendin-4 WOT, on neuronal proliferation and differentiation, and on the metabolism of two neuronal proteins in the rat pheochromocytoma (PC12) cell line, which has been shown to express the GLP-1 receptor. We observed that GLP-1 and exendin-4 induced neurite outgrowth in a manner similar to nerve growth factor (NGF), which was reversed by coincubation with the selective GLP-1 receptor antagonist exendin (9-39). Furthermore, exendin 4 could promote NGF-initiated differentiation and may rescue degenerating cells after NGF-mediated withdrawal. These effects were induced in the absence of cellular dysfunction and toxicity as quantitatively measured by 3-(4,5 cimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide and lactate dehydrogenase assays, respectively. Our findings suggest that such peptides may be used in reversing or halting the neurodegenerative process observed in neurodegenerative diseases, such as the peripheral neuropathy associated with type 2 diabetes mellitus and Alzheimer's and Parkinson's diseases. Due to its novel twin action, GLP-1 and exendin-4 have therapeutic potential for the treatment of diabetic peripheral neuropathy and these central nervous system disorders. PMID- 11861805 TI - Protein kinase C modulation of ethanol inhibition of glycine-activated current in dissociated neurons of rat ventral tegmental area. AB - The brain is particularly sensitive to alcohol during its growth spurt period. To better understand the mechanism(s) involved, we studied the effects of ethanol on neurons freshly dissociated from the ventral tegmental area (VTA) in neonatal rats. Ethanol enhanced (35%) or depressed (45%) glycine-induced responses in VTA neurons (Ye et al., 2001a, 2001b). In this report, we investigated the role of protein kinase C (PKC) and protein kinase A (PKA) in ethanol-induced inhibition of glycine-activated current, using whole-cell patch-clamp technique. Ethanol inhibited glycine-activated current when it was coapplied with the agonist. This inhibition was enhanced when neurons were pretreated with ethanol before the subsequent coapplication of ethanol and glycine. Ethanol's inhibition of glycine activated currents increased with the length of ethanol pretreatment time (ranging from 1 to 30 s), and reached the maximum at 30 s. However, this enhanced inhibition was not seen in the absence of internal ATP. In addition, phorbol-12 myristate-13-acetate (PMA, 100 nM), a PKC activator, markedly inhibited glycine activated current. Blockade of PKC by chelerythrine or by PKC inhibitor peptide significantly attenuated ethanol-induced inhibition. Although partial increase of PKC activity by 1 nM PMA enhanced ethanol inhibition, pretreatment of ethanol did not increase ethanol inhibition after the neurons were treated with 100 nM PMA. These data suggest that ethanol and PKC share the same pathway to suppress glycine receptors. H-89 (1 microM), a selective PKA inhibitor, did not alter glycine-activated current or ethanol inhibition. Our observations suggest that activation of PKC (but not PKA) contributes to ethanol-induced inhibition of glycine receptors. PMID- 11861806 TI - Prevention of antibody-mediated elimination of ligand-targeted liposomes by using poly(ethylene glycol)-modified lipids. AB - One of the major obstacles in the development of ligand-targeted liposomes is poor liposome circulation longevity as a result of antibody-mediated elimination of these highly immunogenic carriers. Because studies from our laboratory suggest that it is not possible to reduce the immunogenicity of ligand-conjugated liposomes by using surface-grafted poly(ethylene glycol) (PEG), we investigated the usefulness of PEG in protecting hapten-conjugated liposomes from elimination by an existing immune response that was previously established against the hapten. Using biotin as a model hapten, a strong biotin-specific antibody response was generated in mice by using bovine serum albumin-biotin. When these animals were challenged with liposomes containing biotin-conjugated lipid (1 or 0.1%), these liposomes were rapidly eliminated. Incorporation of PEG-lipids into these liposomes substantially reduced biotin-specific antibody binding as measured using an in vitro antibody consumption assay. However, depending on the hapten concentration, significant reductions in antibody binding through the use of PEG-lipids may not be sufficient to protect these liposomes from rapid elimination in vivo. Complete protection of liposomes was only achieved when the biotin concentration on liposome surface was low (0.1%) and with 5 mol% of either 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-n-[methoxy(polyethylene glycol) 2000] or 1,2-dipalmatoyl-sn-glycero-3-phosphoethanolamine-n-methoxy(polyethylene glycol)-2000]. The use of 1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine-n [methoxy(polyethylene glycol)-2000] (up to 15 mol%) was not effective in protecting liposomes from rapid elimination in vivo, indicating the limited usefulness of this highly exchangeable PEG-lipid. In conclusion, our in vivo and in vitro data indicate that liposomes can be protected from antibody-mediated elimination by using the right type and concentration of PEG-lipids. This result has important implication in the development of ligand-targeted liposomes. PMID- 11861807 TI - A structure/activity relationship study on arvanil, an endocannabinoid and vanilloid hybrid. AB - Arvanil, a structural "hybrid" between the endogenous cannabinoid CB1 receptor ligand anandamide and capsaicin, is a potent agonist for the capsaicin receptor VR1 (vanilloid receptor type 1), inhibits the anandamide membrane transporter (AMT), and induces cannabimimetic responses in mice. Novel arvanil derivatives prepared by N-methylation, replacement of the amide with urea and thiourea moieties, and manipulation of the vanillyl group were evaluated for their ability to bind/activate CB1 receptors, activate VR1 receptors, inhibit the AMT and fatty acid amide hydrolase (FAAH), and produce cannabimimetic effects in mice. The compounds did not stimulate the CB1 receptor. Methylation of the amide group decreased the activity at VR1, AMT, and FAAH. On the aromatic ring, the substitution of the 3-methoxy group with a chlorine atom or the lack of the 4 hydroxy group decreased the activity on VR1 and AMT, but not the affinity for CB1 receptors, and increased the capability to inhibit FAAH. The urea or thiourea analogs retained activity at VR1 and AMT but exhibited little affinity for CB1 receptors. The urea analog was a potent FAAH inhibitor (IC50 = 2.0 microM). A water-soluble analog of arvanil, O-2142, was as active on VR1, much less active on AMT and CB1, and more potent on FAAH. All compounds induced a response in the mouse "tetrad", particularly those with EC50 <10 nM on VR1. However, the most potent compound, N-N'-di-(3-chloro-4-hydroxy)benzyl-arachidonamide (O-2093, ED50 approximately 0.04 mg/kg), did not activate VR1 or CB1 receptors. Our findings suggest that VR1 and/or as yet uncharacterized receptors produce cannabimimetic responses in mice in vivo. PMID- 11861808 TI - Determinants of ligand selectivity at the kappa-receptor based on the structure of the orphanin FQ receptor. AB - It is unclear how opioid selectivity and activation are regulated within the receptor core. In previous studies, the OFQ receptor was converted into a functional opioid receptor by mutating five amino acids at three sites to the corresponding residues conserved across the mu-, kappa-, and delta-opioid receptors, suggesting that these sites comprise an opioid binding pocket. To examine this hypothesis, the present study examines whether these conserved residues represent an opioid binding pocket in the context of the opioid receptors, i.e., does their removal from opioid receptors destroy opioid ligand binding? The reciprocal mutations K227A (transmembrane [TM]5), IHI290-292VQV (TM6), and I316T (TM7) were evaluated in the kappa-opioid receptor. In terms of alkaloid binding, there were no changes in affinity for mutants K227A and IHI290 292VQV. At mutant I316T, antagonist binding was unaltered, but there was a trend toward slightly decreased agonist affinity. In contrast, the binding of peptides had a more complex pattern. Again, K227A and IHI290-292VQV did not decrease the binding affinity of dynorphin-related peptides. Mutant I316T had 10- to 20-fold decreased affinity for dynorphin-related peptides, suggesting that I316 is part of a critical dynorphin recognition site. In response to alkaloid stimulation, I316T activated more G-protein(s) than wild type, and similar levels were observed in response to dynorphin stimulation. Overall, these results suggest that ligands are capable of achieving high-affinity binding through interaction with multiple sites/conformations of the receptor. These different modes of interaction have different down-stream results in terms of receptor activation and signal transduction. PMID- 11861809 TI - The protein kinase A inhibitor H89 acts on cell morphology by inhibiting Rho kinase. AB - The small GTPase RhoA can retract cell extensions by acting on two Rho kinases (ROCKs). Activated protein kinase A (PKA) inhibits RhoA and induces extensions. The isoquinoline H89 inhibits PKA and thus should prevent the inactivation of RhoA. In kinase assays, H89 has been recently found to inactivate a ROCK-II also. Because H89 may be able to exert opposite effects on cell extensions, we have studied the effects of H89 on neurite formation in the neuroblastoma-glioma line NG 108-15, which expresses ROCK-I and ROCK-II. We found that H89 can indeed inhibit ROCKs and PKA. Because ROCKs act downstream of RhoA, the inhibitory effect of H89 on ROCKs is most prominent. The data indicate that H89 may not be used as an antagonist of PKA in systems in which ROCKs play a role. PMID- 11861810 TI - Amphetamine inhibits the N-methyl-D-aspartate receptor-mediated responses by directly interacting with the receptor/channel complex. AB - Amphetamine (AMPH) induces behavioral sensitization and neurotoxicity primarily by enhancing the dopamine-mediated neurotransmission. However, the involvement of the N-methyl-D-aspartate (NMDA) receptor in AMPH-induced neuropathology is also known. Recent investigation has found that high concentration of dopamine could inhibit NMDA receptor-mediated responses by blocking the NMDA receptor channel. By virtue of the structure similarity between dopamine and AMPH, we determined whether d-AMPH and its analogs, l-AMPH and methamphetamine (MAMH), could affect the NMDA receptor-mediated [3H]N-[1-(2-thienyl)cyclohexyl] piperidine ([3H]TCP) binding in rat cortical membrane preparations and intracellular 45Ca2+ accumulation and cell death in the rat primary cortical cell cultures. AMPH concentration-dependently inhibited NMDA- and glycine-stimulated [3H]TCP binding and intracellular 45Ca2+ accumulation with two distinct potencies; a minor inhibition with high potency and a major inhibition with low potency. [3H]TCP binding suggested that the high-potency inhibition was produced by decreasing agonist-induced activation of the NMDA receptor channel. On the other hand, the low-potency inhibition was produced by competing with [3H]TCP binding in the NMDA receptor channel, like the action of noncompetitive antagonist of the NMDA receptor. However, AMPH analogs were less potent in inhibiting NMDA- and glycine induced cultured cell death. Thus, this result indicates that AMPH could antagonize the NMDA receptor-mediated responses in vitro by two different mechanisms, probably, through directly interacting with two distinct sites on this receptor/channel complex. PMID- 11861811 TI - Daily treatment with diazepam differentially modifies sensitivity to the effects of gamma-aminobutyric acid(A) modulators on schedule-controlled responding in rhesus monkeys. AB - The present study examined how daily treatment with the benzodiazepine (BZ) diazepam modifies the effects of positive modulators acting at different sites on the gamma-aminobutyric acid(A) (GABA(A)) receptor complex and negative modulators acting at BZ sites on the receptor complex. GABA(A) modulators were administered alone or in combination with acute or chronic diazepam to rhesus monkeys (n = 4) responding under a multiple fixed ratio (FR/FR) schedule of food presentation and stimulus-shock termination (SST). There was mutual antagonism between the rate decreasing effects of diazepam (5.6 mg/kg, p.o.) and high efficacy BZ site negative modulators [ethyl beta-carboline-3-carboxylate (beta-CCE), methyl beta carboline-3-carboxylate (beta-CCM) and methyl-6,7-dimethoxyl-4-ethyl-beta carboline-3-carboxylate (DMCM)]. Antagonism of beta-CCE, beta-CCM, and DMCM by diazepam was markedly reduced in monkeys receiving diazepam daily. In contrast, daily diazepam treatment enhanced the rate-decreasing effects of Ro 15-4513 (ethyl 8-azido-6-dihydro-5-methyl-6-oxo-4H-imidazo[1,5-alpha]-[1,4]benzodiazepine 3-carboxylate) and flumazenil. Chronic diazepam elicited cross-tolerance to the BZ triazolam and not to the barbiturate pentobarbital or the neuroactive steroid pregnanolone. These results suggest that tolerance to the rate-decreasing effects of BZs is not accompanied by cross-tolerance to positive GABA(A) modulators acting at other sites on the receptor complex. Moreover, changes in sensitivity to negative GABA(A) modulators during chronic diazepam treatment appeared to be related to negative efficacy and not clearly related to the precipitation of withdrawal for all drugs. These results indicate that changes in sensitivity to the behavioral effects of drugs that act at different sites on the GABA(A) receptor complex might be especially useful for identifying and characterizing the functional consequences of GABA(A) receptor heterogeneity. PMID- 11861812 TI - The platelet-activating factor receptor activates the extracellular signal regulated kinase mitogen-activated protein kinase and induces proliferation of epidermal cells through an epidermal growth factor-receptor-dependent pathway. AB - Platelet-activating factor (PAF) is a lipid mediator that has been implicated in a variety of keratinocyte functions. Keratinocytes express the specific receptor for PAF (PAF-R), a seven-transmembrane G-protein-coupled receptor. Although PAF-R dependent stimulation of numerous signal transduction pathways has been shown in a variety of cell types, to date there has been no analysis of PAF-R signal transduction in human epidermal cells. There is also contradictory evidence that PAF acts as either a suppressor or activator of keratinocyte proliferation. Using a model system created by retroviral-mediated transduction of the PAF-R into the PAF-R-negative epidermal cell line KB, we now demonstrate that the activation of the epidermal PAF-R results in the activation of both the extracellular signal regulated kinase (ERK) and p38, but not the jun N-terminal kinase mitogen activated protein (MAP) kinase pathways. Additionally, we show that the activation of the PAF-R stimulates the replication of epidermal cells. The activation of the ERK signal transduction pathway, as well as the PAF-dependent increase in cell proliferation, was dependent on the transactivation of the epidermal growth factor receptor (EGF-R). PAF-R-induced transactivation of the EGF-R was blocked by pharmacologic inhibitors of matrix metalloproteinases, of heparin-binding epidermal growth factor (HB-EGF), and specific inhibitors of the EGF-R tyrosine kinase. Activation of p38 MAP kinase by the PAF-R was not dependent on EGF-R activation and represents a distinct pathway of PAF-R-mediated signal transduction. In summary, these studies provide a mechanism whereby the PAF-R can exert proliferative effects through the activation of the EGF-R. PMID- 11861814 TI - Peripheral activity of a new NK1 receptor antagonist TAK-637 in the gastrointestinal tract. AB - Pathways controlling gastrointestinal function involve the activation of neurokinin NK1 receptors by substance P (SP) under normal and pathological conditions. Our aim was to pharmacologically characterize the effect of a nonpeptide NK1 receptor antagonist TAK-637 [(aR,9R)-7-[3,5 bis(trifluoromethyl)benzyl]-8,9,10,11-tetrahydro-9-methyl-5-(4-methylphenyl)-7H [1,4]diazocino[2,1-g] [1,7]naphthyridine-6,13-dione] and determine key mechanisms of TAK-637 action in the gastrointestinal tract. Experiments were performed using intestinal preparations isolated from the guinea pig. The selective agonists of NK1 receptors, [Sar9,Met(O2)11]-SP and GR 73632 [H2N-(CH2)4-CO-Phe-Phe-Pro-NMe Leu-Met-NH2], induced contractions in colonic longitudinal muscle pretreated with atropine. TAK-637 (1-100 nM) caused a rightward shift of the concentration response curves showing nanomolar affinity against [Sar9,Met(O2)11]-SP (Kb = 4.7 nM) and GR 73632 (K(b) = 1.8 nM). This antagonist effect remained unchanged by tetrodotoxin. Furthermore, neither the contractions of colonic circular muscle induced by selective activation of NK2 receptors by GR 64349 (Lys-Asp-Ser-Phe-Val Gly-R-gamma-lactam-Leu-Met-NH2) nor the responses of taenia coli induced by the selective NK3 receptor agonist senktide were affected by TAK-637 (100 nM). Studies of electrically induced neurogenic contractions showed that TAK-637 had no effect on cholinergic responses to single-pulse (0.5 ms) stimulation or stimulation with increasing frequency (1-16 Hz, 0.5 ms, 5-s train duration). In contrast, TAK-637 significantly reduced nonadrenergic, noncholinergic contractions of colonic longitudinal muscle evoked at frequencies of 8 to 16 Hz and prevented the development of capsaicin-induced contractions in isolated segments of terminal ileum. Our results indicate that TAK-637 is a selective antagonist of smooth muscle NK(1) receptors that activate intestinal muscle contraction. Additionally TAK-637 inhibits neuronal NK1 receptors involved in the "local" motor response to stimulation of capsaicin-sensitive primary afferents. PMID- 11861813 TI - Unmasking the dynamic interplay between intestinal P-glycoprotein and CYP3A4. AB - Drug efflux by intestinal P-glycoprotein (P-gp) is known to decrease the oral bioavailability of many CYP3A4 substrates. We hypothesized that the interplay occurring between P-gp and CYP3A4 at the apical membrane would increase the opportunity for drug metabolism. To define the roles of P-glycoprotein (P-gp) and CYP3A4 in controlling the extent of intestinal absorption and metabolism, two substrates were tested. The transport, metabolism, and intracellular levels of N methyl piperazine-Phe-homoPhe-vinylsulfone phenyl (K77, a cysteine protease inhibitor; P-gp and CYP3A4 substrate) and felodipine (CYP3A4 substrate only) were measured across CYP3A4-transfected Caco-2 cells in the presence of an inhibitor of CYP3A4 and P-gp, cyclosporine (CsA), or an inhibitor of P-gp and not CYP3A4, GG918 (N-[4-[2-(1,2,3,4-tetrahydro-6,7- dimethoxy-2-isoquinolinyl)-ethyl]-phenyl] 9,10-dihydro-5-methoxy-9-oxo-4-acridine carboxamine). The extent of metabolism was measured by calculating the extraction ratio (ER) across the cells, while accounting for intracellular changes occurring with P-gp inhibition. The (A)pical to (B)asolateral and B-->A ERs for K77 were 0.33 and 0.06, respectively. These changed with GG918 to 0.14 and 0.12 and with CsA to 0.06 and 0.04. Felodipine ERs were similar in both directions, 0.26 and 0.24 (A-->B and B-->A), and were unchanged in the presence of GG918 but decreased with CsA (0.14 and 0.11). The K77 absorption rate was increased 5 and 4.2-fold in the presence of CsA and GG918, respectively, whereas no change was observed for felodipine absorption. The decreased A-->B ER and increased absorption of K77 with GG918 suggest that P gp influences the extent of drug metabolism in the intestine via prolonging the access of drugs to CYP3A4 near the apical membrane and decreasing transport across the cells. PMID- 11861815 TI - Induction of apoptosis by 1,4-benzothiazine analogs in mouse thymocytes. AB - 1,4-benzothiazine (1,4-B) derivatives exert numerous effects in vivo and in vitro, including neurotoxicity and antitumor cytotoxicity. To analyze the mechanisms responsible for 1,4-B-induced cytotoxicity, we performed experiments to evaluate the possible apoptotic effect. For that purpose, we used mouse thymocytes, a cell population well sensitive to induction of apoptosis that has been used to assay apoptosis in many experimental systems. Results indicate that a number of 1,4-B analogs are able to induce both thymocyte apoptosis in vitro and thymus cell loss in vivo. Moreover, analysis of the structure-activity relationship indicate that the sulfur (S) oxidation state, the presence of the carbonyl group, and the nature and position of the side chain modulate the apoptotic efficacy. Moreover, results of in vitro experiments show that the 1,4-B induced apoptosis associates with different biochemical events including phosphatidylcholine-specific phospholipase C activation, acidic sphingomyelinase activation and ceramide generation, loss of mitochondrial membrane potential (DeltaPsi(m)) and cytochrome c release, and caspase-8, -9, and -3 activation. These results indicate that 1,4-B analogs induce apoptosis through a complex of biochemical events. PMID- 11861816 TI - Pharmacokinetic role of P-glycoprotein in oral bioavailability and intestinal secretion of grepafloxacin in vivo. AB - The purpose of this study was to clarify the contribution of P-glycoprotein to the bioavailability and intestinal secretion of grepafloxacin and levofloxacin in vivo. Plasma concentrations of grepafloxacin and levofloxacin after intravenous and intraintestinal administration were increased by cyclosporin A, a P glycoprotein inhibitor, in rats. The total body clearance and volume of distribution at steady state of grepafloxacin were significantly decreased to 60 and 63% of the corresponding control values by cyclosporin A. The apparent oral clearance of grepafloxacin was decreased to 33% of the control, and the bioavailability of grepafloxacin was increased to 95% by cyclosporin A from 53% in the controls. Intestinal clearance of grepafloxacin and levofloxacin were decreased to one-half and one-third of the control, respectively, and biliary clearance of grepafloxacin was also decreased to one-third with cyclosporin A in rats. Intestinal secretion of grepafloxacin in mdr1a/1b (-/-) mice, which lack mdr1-type P-glycoproteins, was significantly decreased compared with wild-type mice, although the biliary secretion was similar. Intestinal secretion of grepafloxacin in wild-type mice treated with cyclosporin A was comparable to those in mdr1a/1b (-/-) mice with or without cyclosporin A, indicating that cyclosporin A completely inhibited P-glycoprotein-mediated intestinal transport of grepafloxacin. In conclusion, our results indicated that P-glycoprotein mediated the intestinal secretion of grepafloxacin and limited the bioavailability of this drug in vivo. PMID- 11861817 TI - Sigma1 receptor modulation of opioid analgesia in the mouse. AB - Opioid analgesia is influenced by many factors, including the sigma1 receptor system. Current studies show the importance of supraspinal mechanisms in these sigma1actions. Given supraspinally, the sigma1receptor agonist (+)pentazocine diminished systemic mu, delta, kappa1, and kappa3 opioid analgesia in CD-1 mice. There was a trend for the kappa drugs to be more sensitive to the fixed dose of (+)pentazocine, although the differences did not achieve statistical significance. In contrast to its actions supraspinally, (+)pentazocine was without effect against morphine when both were given spinally. These findings are consistent with a supraspinal site of anti-opioid action of (+)pentazocine. Down regulating supraspinal sigma1binding sites using an antisense approach potentiated mu, delta, kappa1, and kappa3 analgesia in CD-1 mice. Although equally responsive to mu drugs, BALB-c mice are far less sensitive to kappa analgesics than CD-1 mice. Earlier studies reported that these different responses to kappa drugs between CD-1 and BALB-c were eliminated by the concurrent administration of haloperidol, a sigma1 antagonist. Antisense treatment of BALB-c mice markedly enhanced the response to kappa drugs, as well as morphine. This enhanced response following antisense treatment was similar to that seen with haloperidol. These observations confirm the importance of sigma1 receptors as a modulatory system influencing the analgesic activity of opioid drugs. PMID- 11861818 TI - Methotrexate exacerbates tumor progression in a murine model of chronic myeloid leukemia. AB - Expression of drug-resistant forms of dihydrofolate reductase (DHFR) in hematopoietic cells confers substantial resistance of animals to antifolate administration. In this study, we tested whether the chemoprotection conferred by expression of the tyrosine-22 variant DHFR could be used for more effective therapy of the 32Dp210 murine model of chronic myeloid leukemia (CML). 32Dp210 tumor cells were found to be sensitive to methotrexate (MTX) in vitro, whereas cells expressing the tyrosine-22 DHFR gene were protected from MTX at up to micromolar concentrations. MTX administered at low dose (2 mg/kg/day) did not protect normal C3H-He/J mice from 32Dp210 tumor infused intravenously, with drug toxicity limiting the administration of higher doses. Animals engrafted with transgenic tyrosine-22 DHFR marrow were protected from greater MTX doses (up to 6 mg/kg/day). However, the increased doses of MTX afforded by drug-resistance gene expression surprisingly resulted in decreased survival of the transplanted tumor bearing animals, with increased levels of tumor detected in peripheral blood. This apparent exacerbation of tumor progression by MTX was not observed in DHFR transgenic mice in which all cells and tissues contain the drug-resistance gene. This suggests that increased tumor progression in MTX-administered animals resulted from MTX sensitivity of a nonhematopoietic host component, thus allowing tumor expansion. We conclude that MTX exacerbates tumor progression in the 32Dp210 model of CML, and that based on this model alternate DHFR inhibitors combined with drug-resistant DHFR or other chemotherapeutic agent/drug-resistance gene combinations may be required for the application of drug-resistance gene expression to the treatment of CML. PMID- 11861819 TI - Regulation of corticotropin-releasing factor neuronal systems and hypothalamic pituitary-adrenal axis activity by stress and chronic antidepressant treatment. AB - In a series of experiments, we tested the hypothesis that chronic antidepressant drug administration reduces the synaptic availability of corticotropin-releasing factor (CRF) through one or more effects on CRF gene expression or peptide synthesis. We also determined whether effects of acute or chronic stress on CRF gene expression or peptide concentration are influenced by antidepressant drug treatment. Four-week treatment with venlafaxine, a dual serotonin (5 HT)/norepinephrine (NE) reuptake inhibitor, and tranylcypromine, a monoamine oxidase inhibitor, resulted in an attenuation of acute stress-induced increases in CRF heteronuclear RNA (hnRNA) synthesis in the paraventricular nucleus (PVN). Trends toward the same effect were observed after treatment with the 5-HT reuptake inhibitor fluoxetine, or the NE reuptake inhibitor reboxetine. CRF mRNA accumulation in the PVN during exposure to chronic variable stress was attenuated by concurrent antidepressant administration. Basal CRF hnRNA and mRNA expression were not affected by antidepressant treatment in the PVN or in other brain regions examined. Chronic stress reduced CRF concentrations in the median eminence, but there were no consistent effects of antidepressant drug treatment on CRF, serum corticotropin, or corticosterone concentrations. CRF receptor expression and basal and stress-stimulated HPA axis activity were unchanged after antidepressant administration. These results suggest that chronic antidepressant administration diminishes the sensitivity of CRF neurons to stress rather than alters their basal activity. Additional studies are required to elucidate the functional consequences and mechanisms of this interaction. PMID- 11861821 TI - Signals of oxidant-induced cardiomyocyte hypertrophy: key activation of p70 S6 kinase-1 and phosphoinositide 3-kinase. AB - Cardiomyocytes in culture can survive low or mild doses of oxidants but later increase cell volume and protein content. To understand the mechanism, we determined the early signaling events of oxidative stress. With 200 microM H2O2, the activity of p70 S6 kinase-1 (p70S6K1) increased at 30 min and reached a plateau at 90 min. Dose-response studies at the 60 min time point show that p70S6K1 activity reached its highest level with 150 microM H2O2. Increased p70S6K1 activity correlated with phosphorylation of Thr389 and Thr421/Ser424 residues, suggesting the involvement of an upstream kinase. Phosphoinositide 3 kinase (PI3K) activity was elevated by 5 min, reached a plateau at 10 min, and remained more than 6-fold induced for at least 60 min after 200 microM H2O2 exposure. The dose-response studies at 10 min found that 150 microM H2O2 induced the highest PI3K activity. Increased PI3K activity correlated with tyrosine phosphorylation of the 85-kDa regulatory subunit. Inactivating PI3K with wortmannin prevented H2O2 from inducing Thr389 phosphorylation and p70S6K1 activation. Wortmannin and rapamycin prevented H2O2 from inducing increases in cell volume and protein content. The antineoplastic drugs doxorubicin and daunorubicin also induced significant enlargement of cardiomyocytes at 10 to 100 nM dose range. Although the glutathione synthesis inhibitor buthionine sulfoximine potentiated the effect of doxorubicin and H2O2, the antioxidant N acetylcysteine prevented induction of cell enlargement. Our data suggest that oxidative stress induces activation of PI3K, which leads to p70S6K1 activation and enlargement of cell size. PMID- 11861820 TI - Methylenedioxymethamphetamine decreases plasmalemmal and vesicular dopamine transport: mechanisms and implications for neurotoxicity. AB - Administration of a high-dose regimen of methamphetamine (METH) rapidly and profoundly decreases plasmalemmal and vesicular dopamine (DA) transport in the striatum, as assessed in synaptosomes and purified vesicles, respectively. To determine whether these responses were common to other amphetamines of abuse, effects of methylenedioxymethamphetamine (MDMA) on the plasmalemmal DA transporter (DAT) and vesicular monoamine transporter-2 (VMAT-2) were assessed. Similar to effects of METH reported previously, multiple high-dose MDMA administrations rapidly (within 1 h) decreased plasmalemmal DA uptake, as assessed ex vivo in synaptosomes prepared from treated rats. Unlike effects of multiple METH injections, this deficit was reversed completely 24 h after drug treatment. Also in contrast to effects of multiple METH injections, 1) MDMA caused little or no decrease in binding of the DAT ligand WIN35428, and 2) neither prevention of hyperthermia nor prior depletion of DA prevented the MDMA induced reduction in plasmalemmal DA transport. However, a role for phosphorylation was suggested because pretreatment with protein kinase C inhibitors attenuated the deficit caused by MDMA in an in vitro model system. In addition to affecting DAT function, MDMA rapidly decreased vesicular DA transport as assessed in striatal vesicles prepared from treated rats. Unlike effects of multiple METH injections reported previously, this decrease partially recovered by 24 h after drug treatment. Taken together, these results reveal several differences between effects of MDMA and previously reported METH on DAT and VMAT 2; differences that may underlie the dissimilar neurotoxic profile of these agents. PMID- 11861822 TI - Effects of mu-opioid agonists on cocaine- and food-maintained responding and cocaine discrimination in rhesus monkeys: role of mu-agonist efficacy. AB - Mu-opioid agonists decrease cocaine self-administration in laboratory studies and cocaine use by many cocaine- and opioid-dependent polydrug abusers. To assess the role of mu-agonist efficacy as a determinant of these effects, this study evaluated cocaine- and food-maintained responding by rhesus monkeys (Macaca mulatta) during chronic treatment with saline or the high-efficacy mu-agonist fentanyl (0.001-0.01 mg/kg/h), the intermediate-efficacy mu-agonist morphine (0.032-0.32 mg/kg/h), or the low-efficacy mu-agonists nalbuphine (0.1-1.0 mg/kg/h) and butorphanol (0.0032-0.032 mg/kg/h). Responding was maintained by cocaine and food under a second order schedule of reinforcement during multiple daily sessions of cocaine and food availability. Saline and each opioid dose were administered continuously for 7 consecutive days during availability of each cocaine dose. All four mu-agonists produced dose-dependent and sustained decreases in cocaine self-administration across a range of cocaine doses (0.0032 0.1 mg/kg/injection). Nalbuphine and butorphanol produced the greatest decreases in cocaine self-administration and the smallest effects on food-maintained responding. Morphine and fentanyl produced smaller decreases in cocaine self administration, and undesirable effects precluded evaluation of higher fentanyl and morphine doses. Decreases in cocaine self-administration produced by nalbuphine and butorphanol probably did not reflect a general blockade of cocaine's abuse-related effects, because nalbuphine and butorphanol did not block the discriminative stimulus effects of cocaine in monkeys trained to discriminate 0.4 mg/kg cocaine from saline in a food-reinforced drug discrimination procedure. These results suggest that low-efficacy mu-agonists may decrease cocaine self administration to a greater degree and with fewer undesirable effects than high efficacy mu-agonists. PMID- 11861823 TI - Characterization of (2S,4R)-1-[5-chloro-1-[(2,4-dimethoxyphenyl)sulfonyl]-3-(2 methoxy-phenyl)-2-oxo-2,3-dihydro-1H-indol-3-yl]-4-hydroxy-N,N-dimethyl-2 pyrrolidine carboxamide (SSR149415), a selective and orally active vasopressin V1b receptor antagonist. AB - (2S,4R)-1-[5-Chloro-1-[(2,4-dimethoxyphenyl)sulfonyl]-3-(2-methoxy-phenyl)-2-oxo 2,3-dihydro-1H-indol-3-yl]-4-hydroxy-N,N-dimethyl-2-pyrrolidine carboxamide (SSR149415), the first selective, nonpeptide vasopressin V1b receptor antagonist yet described, has been characterized in vitro and in vivo. SSR149415 showed competitive nanomolar affinity for animal and human V1b receptors and exhibited much lower affinity for rat and human V1a, V2, and oxytocin receptors. Moreover, this compound did not interact with a large number of other receptors, enzymes, or ion channels. In vitro, SSR149415 behaved as a full antagonist and potently inhibited arginine vasopressin (AVP)-induced Ca2+ increase in Chinese hamster ovary cells expressing rat or human V1b receptors. The in vivo activity of SSR149415 has been studied in several models of elevated corticotropin secretion in conscious rats. SSR149415 inhibited exogenous AVP-induced increase in plasma corticotropin, from 3 mg/kg i.p. and 10 mg/kg p.o. upwards. Similarly, this compound antagonized AVP-potentiated corticotropin release provoked by exogenous corticoliberin at 3 mg/kg p.o. The effect lasted for more than 4 h at 10 mg/kg p.o. showing a long-lasting oral effect. SSR149415 (10 mg/kg p.o.) also blocked corticotropin secretion induced by endogenous AVP increase subsequent to body water loss. Moreover, 10 mg/kg i.p SSR149415 inhibited plasma corticotropin elevation after restraint-stress in rats by 50%. In the four-plate test, a mouse model of anxiety, SSR149415 (3 mg/kg p.o. upwards) displayed anxiolytic-like activity after acute and 7-day repeated administrations. Thus, SSR149415 is a potent, selective, and orally active V1b receptor antagonist. It represents a unique tool for exploring the functional role of V1b receptors and deserves to be clinically investigated in the field of stress and anxiety. PMID- 11861824 TI - Reovirus-induced alterations in gene expression related to cell cycle regulation. AB - Mammalian reovirus infection results in perturbation of host cell cycle progression. Since reovirus infection is known to activate cellular transcription factors, we investigated alterations in cell cycle-related gene expression following HEK293 cell infection by using the Affymetrix U95A microarray. Serotype 3 reovirus infection results in differential expression of 10 genes classified as encoding proteins that function at the G(1)-to-S transition, 11 genes classified as encoding proteins that function at G(2)-to-M transition, and 4 genes classified as encoding proteins that function at the mitotic spindle checkpoint. Serotype 1 reovirus infection results in differential expression of four genes classified as encoding proteins that function at the G(1)-to-S transition and three genes classified as encoding proteins that function at G(2)-to-M transition but does not alter any genes classified as encoding proteins that function at the mitotic spindle checkpoint. We have previously shown that serotype 3, but not serotype 1, reovirus infection induces a G(2)-to-M transition arrest resulting from an inhibition of cdc2 kinase activity. Of the differentially expressed genes encoding proteins regulating the G(2)-to-M transition, chk1, wee1, and GADD45 are known to inhibit cdc2 kinase activity. A hypothetical model describing serotype 3 reovirus-induced inhibition of cdc2 kinase is presented, and reovirus-induced perturbations of the G(1)-to-S, G(2)-to-M, and mitotic spindle checkpoints are discussed. PMID- 11861825 TI - CD4 independence of simian immunodeficiency virus Envs is associated with macrophage tropism, neutralization sensitivity, and attenuated pathogenicity. AB - To investigate the basis for envelope (Env) determinants influencing simian immunodeficiency virus (SIV) tropism, we studied a number of Envs that are closely related to that of SIVmac239, a pathogenic, T-tropic virus that is neutralization resistant. The Envs from macrophage-tropic (M-tropic) virus strains SIVmac316, 1A11, 17E-Fr, and 1100 facilitated infection of CCR5-positive, CD4-negative cells. In contrast, the SIVmac239 Env was strictly dependent upon the presence of CD4 for membrane fusion. We also found that the Envs from M tropic virus strains, which are less pathogenic in vivo, were very sensitive to antibody-mediated neutralization. Antibodies to the V3-loop, as well as antibodies that block SIV gp120 binding to CCR5, efficiently neutralized CD4 independent, M-tropic Envs but not the 239 Env. However, triggering the 239 Env with soluble CD4, presumably resulting in exposure of the CCR5 binding site, made it as neutralization sensitive as the M-tropic Envs. In addition, mutations of N linked glycosylation sites in the V1/V2 region, previously shown to enhance antigenicity and immunogenicity, made the 239 Env partially CD4 independent. These findings indicate that Env-based determinants of M tropism of these strains are generally associated with decreased dependence on CD4 for entry into cells. Furthermore, CD4 independence and M tropism are also associated with neutralization sensitivity and reduced pathogenicity, suggesting that the humoral immune response may exert strong selective pressure against CD4-independent M tropic SIVmac strains. Finally, genetic modification of viral Envs to enhance CD4 independence may also result in improved humoral immune responses. PMID- 11861827 TI - Interferon-regulated pathways that control hepatitis B virus replication in transgenic mice. AB - We previously showed that the intrahepatic induction of cytokines such as alpha/beta interferon (IFN-alpha/beta) and gamma interferon (IFN-gamma) inhibits hepatitis B virus (HBV) replication noncytopathically in the livers of transgenic mice. The intracellular pathway(s) responsible for this effect is still poorly understood. To identify interferon (IFN)-inducible intracellular genes that could play a role in our system, we crossed HBV transgenic mice with mice deficient in IFN regulatory factor 1 (IRF-1), the double-stranded RNA-activated protein kinase (PKR), or RNase L (RNase L) (IRF-1(-/-), PKR(-/-), or RNase L(-/-) mice, respectively), three well-characterized IFN-inducible genes that mediate antiviral activity. We showed that unmanipulated IRF-1(-/-) or PKR(-/-) transgenic mice replicate HBV in the liver at slightly higher levels than the respective controls, suggesting that both IRF-1 and PKR individually appear to mediate signals that modulate HBV replication under basal conditions. These same animals were responsive to the antiviral effects of the IFN-alpha/beta inducer poly(I-C) or recombinant murine IFN-gamma, suggesting that under these conditions, either the IRF-1 or the PKR genes can mediate the antiviral activity of the IFNs or other IFN-inducible genes mediate the antiviral effects. Finally, RNase L(-/-) transgenic mice were undistinguishable from controls under basal conditions and after poly(I-C) or IFN-gamma administration, suggesting that RNase L does not modulate HBV replication in this model. PMID- 11861826 TI - Enhancing the proteolytic maturation of human immunodeficiency virus type 1 envelope glycoproteins. AB - In virus-infected cells, the envelope glycoprotein (Env) precursor, gp160, of human immunodeficiency virus type 1 is cleaved by cellular proteases into a fusion-competent gp120-gp41 heterodimer in which the two subunits are noncovalently associated. However, cleavage can be inefficient when recombinant Env is expressed at high levels, either as a full-length gp160 or as a soluble gp140 truncated immediately N-terminal to the transmembrane domain. We have explored several methods for obtaining fully cleaved Env for use as a vaccine antigen. We tested whether purified Env could be enzymatically digested with purified protease in vitro. Plasmin efficiently cleaved the Env precursor but also cut at a second site in gp120, most probably the V3 loop. In contrast, a soluble form of furin was specific for the gp120-gp41 cleavage site but cleaved inefficiently. Coexpression of Env with the full-length or soluble form of furin enhanced Env cleavage but also reduced Env expression. When the Env cleavage site (REKR) was mutated in order to see if its use by cellular proteases could be enhanced, several mutants were found to be processed more efficiently than the wild-type protein. The optimal cleavage site sequences were RRRRRR, RRRRKR, and RRRKKR. These mutations did not significantly alter the capacity of the Env protein to mediate fusion, so they have not radically perturbed Env structure. Furthermore, unlike that of wild-type Env, expression of the cleavage site mutants was not significantly reduced by furin coexpression. Coexpression of Env cleavage site mutants and furin is therefore a useful method for obtaining high level expression of processed Env. PMID- 11861828 TI - Envelope gene-mediated neurovirulence in feline immunodeficiency virus infection: induction of matrix metalloproteinases and neuronal injury. AB - The release of neurotoxins by activated brain macrophages or microglia is one mechanism proposed to contribute to the development of neurological disease following infection by lentiviruses, including feline immunodeficiency virus (FIV). Since molecular diversity in the lentiviral envelope gene influences the expression of host molecules implicated in neuronal injury, the role of the envelope sequence in FIV neuropathogenesis was investigated by using the neurovirulent FIV strain V1CSF, the nonneurovirulent strain Petaluma, and a chimera (FIVCh) containing the V1CSF envelope gene in a Petaluma background. All three viruses replicated in primary feline macrophages with equal efficiency, but conditioned medium from V1CSF- or FIVCh-infected cells was significantly more neurotoxic than medium from Petaluma-infected cultures (P < 0.001) and could be attenuated in a dose-dependent manner by treatment with either the matrix metalloproteinase (MMP) inhibitor prinomastat (PMT) or function-blocking antibodies to MMP-2. Although FIV sequences were detectable by PCR in brain tissue from neonatal cats infected with each of the viral strains, immunohistochemistry revealed increased astrogliosis and macrophage activation in the brains of V1CSF- and FIVCh-infected cats relative to the other groups, together with elevated markers of neuronal stress that included morphological changes and increased c-fos immunoreactivity. Similarly, MMP-2, but not MMP-9, mRNA and protein expression was increased in brain tissues of V1CSF- and FIVCh infected cats relative to Petaluma-infected animals (P < 0.01). Infection with V1CSF or FIVCh was also associated with greater CD4(+) cell depletion (P < 0.001) and neurodevelopmental delays (P < 0.005), than in Petaluma-infected animals; these deficits improved following PMT therapy. These findings indicated that diversity in the envelope gene sequence influenced the neurovirulence exhibited by FIV both in vitro and in vivo, possibly through a mechanism involving the differential induction of MMP-2. PMID- 11861829 TI - Identification of Kaposi's sarcoma-associated herpesvirus (KSHV)-specific cytotoxic T-lymphocyte epitopes and evaluation of reconstitution of KSHV-specific responses in human immunodeficiency virus type 1-Infected patients receiving highly active antiretroviral therapy. AB - Following the introduction of highly active antiretroviral therapy (HAART), the incidence of Kaposi's sarcoma (KS) has significantly declined in human immunodeficiency virus type 1 (HIV-1)-positive (HIV-1(+)) individuals and clinical remission is often observed. We hypothesize that these effects are partly due to anti-KS-associated herpesvirus (KSHV) immune restoration. Here, 15 mer overlapping peptides from proteins K12 and K8.1 were used to identify novel KSHV-specific cytotoxic T-lymphocyte epitopes. Three immunogenic peptides, two lytic and one latent, were subsequently used to monitor the anti-KSHV CD8(+) T cell responses in a cohort of 19 HIV-1(+) KSHV(+/-) KS(+/-) individuals during 52 weeks of HAART. KSHV and HIV-1 loads, KSHV antibody titers, and both CD4(+) and CD8(+) T-lymphocyte counts were enumerated. Prior to HAART, the total number of spot-forming cells (SFC) for all three peptides correlated with both CD4(+) and CD8(+) T-lymphocyte counts (P < or = 0.05) in the KSHV-positive KS-positive cohort (n = 11). Following 52 weeks of HAART, significant decreases in HIV-1 and KSHV loads were associated with significant increases in CD4(+) T-lymphocyte counts and number of SFC for the three KSHV-specific peptides. Although these increases were modest in comparison to the number of SFC observed with the HIV-1 gag peptide SLYNTVATL, they represented a fourfold increase from the baseline, continuing an upward trend to week 52. PMID- 11861830 TI - Budding of equine infectious anemia virus is insensitive to proteasome inhibitors. AB - The only retrovirus protein required for the budding of virus-like particles is the Gag protein; however, recent studies of Rous sarcoma virus (RSV) and human immunodeficiency virus have suggested that modification of Gag with ubiquitin (Ub) is also required. As a consequence, the release of these viruses is reduced in the presence of proteasome inhibitors, which indirectly reduce the levels of free Ub within the cell. Here we show that the budding of equine infectious anemia virus (EIAV) from infected equine cells is largely unaffected by these drugs, although use of one inhibitor (MG-132) resulted in a dramatic block to proteolytic processing of Gag. This lack of sensitivity was also observed in transiently transfected avian cells under conditions that greatly reduce RSV budding. Moreover, insensitivity was observed when the EIAV Gag protein was expressed in the absence of all the other virus products, indicating that they are not required for this phenotype. An activity that enables EIAV to tolerate exposure to proteasome inhibitors was mapped to the C-terminal p9 sequence, as demonstrated by the ability of an RSV Gag-p9 chimera to bud in the presence of the drugs. Intriguingly, the p9 sequence contains a short sequence motif that is similar to a surface-exposed helix of Ub, suggesting that EIAV Gag may have captured a function that allows it to bypass the need for ubiquitination. Thus, the mechanism of EIAV budding may not be substantially different from that of other retroviruses, even though it behaves differently in the presence of proteasome inhibitors. PMID- 11861831 TI - Human T-cell leukemia virus type 2 (HTLV-2) Tax protein transforms a rat fibroblast cell line but less efficiently than HTLV-1 Tax. AB - Human T-cell leukemia virus type 1 (HTLV-1) and HTLV-2 are retroviruses with similar biological properties. Whereas HTLV-1 is the causative agent of an aggressive T-cell leukemia, HTLV-2 has been associated with only a few cases of lymphoproliferative disorders. Tax1 and Tax2 are the transcriptional activators of HTLV-1 and HTLV-2, respectively. Here we show that Tax2 transformed a Rat-1 fibroblast cell line to form colonies in soft agar, but the size and number of the colonies were lower than those of Tax1. Use of a chimeric Tax protein showed that the C-terminal amino acids 300 to 353 were responsible for the high transforming activity of Tax1. Activation of cellular genes by Tax1 through transcription factor NF-kappa B is reportedly essential for the transformation of Rat-1 cells. Tax2 also activated the transcription through NF-kappa B in Rat-1 cells, and such activity was equivalent to that induced by Tax1. Thus, the high transforming activity of Tax1 is mediated by mechanisms other than NF-kappa B activation. Our results showed that Tax2 has a lower transforming activity than Tax1 and suggest that the high transforming activity of Tax1 is involved in the leukemogenic property of HTLV-1. PMID- 11861832 TI - Repression of African swine fever virus polyprotein pp220-encoding gene leads to the assembly of icosahedral core-less particles. AB - African swine fever virus (ASFV) polyprotein pp220, encoded by the CP2475L gene, is an N-myristoylated precursor polypeptide that, after proteolytic processing, gives rise to the major structural proteins p150, p37, p34, and p14. These proteins localize at the core shell, a matrix-like virus domain placed between the DNA-containing nucleoid and the inner envelope. In this study, we have examined the role of polyprotein pp220 in virus morphogenesis by means of an ASFV recombinant, v220i, containing an inducible copy of the CP2475L gene regulated by the Escherichia coli repressor-operator system. Under conditions that repress pp220 expression, the virus yield of v220i was about 2.6 log units lower than that of the parental virus or of the recombinant grown under permissive conditions. Electron microscopy revealed that pp220 repression leads to the assembly of icosahedral particles virtually devoid of the core structure. Analysis of recombinant v220i by immunoelectron microscopy, immunoblotting, and DNA hybridization showed that mutant particles essentially lack, besides the pp220-derived products, a number of major core proteins as well as the viral DNA. On the other hand, transient expression of the CP2475L gene in COS cells showed that polyprotein pp220 assembles into electron-dense membrane-bound coats, whereas a mutant nonmyristoylated version of pp220 does not associate with cellular membranes but forms large cytoplasmic aggregates. Together, these findings indicate that polyprotein pp220 is essential for the core assembly and suggest that its myristoyl moiety may function as a membrane-anchoring signal to bind the developing core shell to the inner viral envelope. PMID- 11861833 TI - Novel, chimpanzee serotype 68-based adenoviral vaccine carrier for induction of antibodies to a transgene product. AB - An E1-deletion-containing adenoviral recombinant based on the chimpanzee serotype 68 (AdC68) was developed to express the rabies virus glycoprotein. Mice immunized with this construct (AdC68rab.gp) developed antibodies to rabies virus and remained resistant to challenge with an otherwise lethal dose of rabies virus. In naive mice immunized intranasally, the rabies virus-specific antibody responses elicited by AdC68rab.gp were comparable with regard to both titers and isotype profiles to those induced by an adenoviral recombinant based on human serotype 5 (Adhu5) expressing the same transgene product. In contrast, subcutaneous immunization with the AdC68rab.gp vaccine resulted in markedly lower antibody responses to the rabies virus glycoprotein than the corresponding Adhu5 vaccine. Antibodies from AdC68rab.gp-immunized mice were strongly biased towards the immunoglobulin G2a isotype. The antibody response to the rabies virus glycoprotein presented by Adhu5rab.gp was severely compromised in animals preexposed to the homologous adenovirus. In contrast, the rabies virus-specific antibody response to the AdC68rab.gp vaccine was at most marginally affected by preexisting immunity to common human adenovirus serotypes, such as 2, 4, 5, 7, and 12. This novel vaccine carrier thus offers a distinct advantage over adenoviral vaccines based on common human serotypes. PMID- 11861834 TI - Improving DNA vaccine potency by linking Marek's disease virus type 1 VP22 to an antigen. AB - We have previously employed an intercellular spreading strategy using herpes simplex virus type 1 (HSV-1) VP22 protein to enhance DNA vaccine potency because DNA vaccines lack the intrinsic ability to amplify in cells. Recently, studies have demonstrated that the protein encoded by UL49 of Marek's disease virus type 1 (MDV-1) exhibits some degree of homology to the HSV-1 VP22 protein and features the property of intercellular transport. We therefore generated a DNA vaccine encoding MDV-1 VP22 linked to a model antigen, human papillomavirus type 16 E7. We demonstrated that compared with mice vaccinated with DNA encoding wild-type E7, mice vaccinated with MDV-1 VP22/E7 DNA exhibited a significant increase in number of gamma-interferon-secreting, E7-specific CD8(+)-T-cell precursors as well as stronger tumor prevention and treatment effects. Furthermore, our data indicated that the antitumor effect was CD8 dependent. These results suggested that the development of vaccines encoding VP22 fused to a target antigen might be a promising strategy for improving DNA vaccine potency. PMID- 11861835 TI - Truncation of the cytoplasmic domain induces exposure of conserved regions in the ectodomain of human immunodeficiency virus type 1 envelope protein. AB - We have described a CD4-independent variant of HXBc2, termed 8x, that binds directly to CXCR4 and mediates CD4-independent virus infection. Determinants for CD4 independence map to residues in the V3 and V4-C4 domains together with a single nucleotide deletion in the transmembrane domain which introduces a frameshift (FS) at position 706. This FS results in a truncated cytoplasmic domain of 27 amino acids. We demonstrate here that while introduction of the 8x FS mutation into heterologous R5, X4, or R5X4 Env proteins did not impart CD4 independence, it did affect the conformation of the gp120 surface subunit, exposing highly conserved domains involved in both coreceptor and CD4 binding. In addition, antigenic changes in the gp41 ectodomain were also observed, consistent with the idea that the effects of cytoplasmic domain truncation must in some way be transmitted to the external gp120 subunit. Truncation of gp41 also resulted in the marked neutralization sensitivity of all Env proteins tested to human immunodeficiency virus-positive human sera and monoclonal antibodies directed against the CD4 or coreceptor-binding sites. These results demonstrate a structural interdependence between the cytoplasmic domain of gp41 and the ectodomain of the Env protein. They also may help explain why the length of the gp41 cytoplasmic domain is retained in vivo and may provide a way to genetically trigger the exposure of neutralization determinants in heterologous Env proteins that may prove useful for vaccine development. PMID- 11861836 TI - Human immunodeficiency virus type 1 Nef binds to tumor suppressor p53 and protects cells against p53-mediated apoptosis. AB - The nef gene product of human immunodeficiency virus type 1 (HIV-1) is important for the induction of AIDS, and key to its function is its ability to manipulate T cell function by targeting cellular signal transduction proteins. We reported that Nef coprecipitates a multiprotein complex from cells which contains tumor suppressor protein p53. We now show that Nef interacts directly with p53. Binding assays showed that an N-terminal, 57-residue fragment of Nef (Nef 1-57) contains the p53-binding domain. Nef also interacted with p53 during HIV-1 infection in vitro. As p53 plays a critical role in the regulation of apoptosis, we hypothesized that Nef may alter this process. Nef inhibited UV light-induced, p53 dependent apoptosis in MOLT-4 cells, with Nef 1-57 being as effective as its full length counterpart. The inhibition by Nef of p53 apoptotic function is most likely due its observed ability to decrease p53 protein half-life and, consequently, p53 DNA binding activity and transcriptional activation. These data show that HIV-1 Nef may augment HIV replication by prolonging the viability of infected cells by blocking p53-mediated apoptosis. PMID- 11861837 TI - Mouse model for the equilibration interaction between the host immune system and human T-cell leukemia virus type 1 gene expression. AB - To study the involvement of immune responses against Tax of human T-cell leukemia virus type 1 (HTLV-1) in the growth of and gene suppression in Tax-expressing tumor cells in vivo, we established a model system involving C57BL/6J mice and a syngeneic lymphoma cell line, EL4. When mice were immunized by DNA-based immunization with Tax expression plasmids, solid tumor formation upon subcutaneous inoculation of EL4 cells expressing green fluorescent protein-fused Tax (Gax) under the control of the HTLV-1 enhancer was strongly inhibited, and in vitro analysis showed that DNA immunization elicited cytotoxic T-lymphocyte (CTL) responses but not production of antibodies to Tax protein. Since EL4/Gax cells inoculated into DNA-immunized mice were not completely eradicated but were maintained as small solid tumors for a long period, there appeared to be a certain equilibrium between CTL activity and the growth of Gax-expressing cells. With such a balance, expression of the Gax gene in EL4/Gax cells was strongly suppressed. These results suggested that gene expression under the control of the HTLV-1 long terminal repeat and Tax is silenced in vivo, resulting in an equilibrium between viral expression and the host immune system. Such a balance would represent a status of persistent infection by HTLV-1 in virus-infected individuals during the latency period. PMID- 11861838 TI - Characterization of chromosomally assigned replication-competent gamma porcine endogenous retroviruses derived from a large white pig and expression in human cells. AB - Vertically transmitted endogenous retroviruses pose an infectious risk in the course of pig-to-human transplantation of cells, tissues, and organs. Two classes of polytropic type C porcine endogenous retroviruses (PERV) productively infect human cells in vitro. The cloning and characterization of replication-competent PERV-B sequences from infected human cells (F. Czauderna, N. Fischer, K. Boller, R. Kurth, and R. R. Tonjes, J. Virol. 74:4028-4038, 2000) as well as the cloning of functional PERV-A and -B sequences from porcine cell line PK15 (U. Krach, N. Fischer, F. Czauderna, and R. R. Tonjes, J. Virol. 75:5465-5472, 2001) have been previously described. Here we report the isolation of four full-length proviral sequences from a porcine bacterial artificial chromosome (BAC) library that comprises chromosomally assigned PERV. Clones Bac-PERV-A(130A12) and Bac-PERV A(151B10) map to pig chromosome 1 and demonstrate close homology to PK15-PERV A(58) in env and to PERV-MSL in long terminal repeat (LTR), gag, and pro/pol sequences. Clone Bac-PERV-A(463H12) is located on pig chromosome 3 and demonstrates close homology to PK15-PERV-A(58) in env and to 293-PERV-B(43) in LTR, gag, and pro/pol (Czauderna et al.; R. R. Tonjes, F. Czauderna, N. Fischer, U. Krach, K. Boller, P. Chardon, C. Rogel-Gailard, M. Niebert, G. Scheef, A. Werner, and R. Kurth, Transplant Proc. 32:1158-1161, 2000). Clone Bac-PERV B(192B9) is located on pig chromosome 7 in the swine leukocyte antigen region and is highly homologous with but distinct from the previously described functional clone 293-PERV-B(43) and bears the number of repeats initially observed in the LTRs of clone 293-PERV-A(42) (Czauderna et al.; Krach et al.). Clones Bac-PERV A(130A12), Bac-PERV-A(151B10), and Bac-PERV-A(463H12) were replication competent upon transfection into susceptible 293 and HeLa cells. Bac-PERV-B(192B9), however, bears two stop codons in pro/pol preventing this clone from being replication competent in some individual pigs, but initial screenings indicate that this provirus might be intact in others. The data suggest that the porcine genome harbors a limited number of infectious PERV sequences, allowing for specific screening in different pig breeds. PMID- 11861839 TI - Interaction between STAT-3 and HNF-3 leads to the activation of liver-specific hepatitis B virus enhancer 1 function. AB - The signal transducer and activator of transcription 3 (STAT-3), a member of the STAT family of proteins, binds to a large number of transcriptional control elements and regulates gene expression in response to cytokines. While it binds to its cognate nucleotide sequences, it has been recently shown to directly interact with other transcriptional factors in the absence of DNA. We report here one such novel interaction between STAT-3 and hepatocyte nuclear factor 3 (HNF-3) in the absence of DNA. We have identified a STAT-3 binding site within the core domain of hepatitis B virus (HBV) enhancer 1. The HBV enhancer 1 DNA-STAT-3 protein interaction is shown to be stimulated by interleukin-6 (IL-6) and epidermal growth factor, which leads to an overall stimulation of HBV enhancer 1 function and viral gene expression. Using mobility shift assays and transient transfection schemes, we demonstrate a cooperative interaction between HNF-3 and STAT-3 in mediating the cytokine-mediated HBV enhancer function. Cytokine stimulation of HBV gene expression represents an important regulatory scheme of direct relevance to liver disease pathogenesis associated with HBV infection. PMID- 11861841 TI - Specific inactivation of inhibitory sequences in the 5' end of the human papillomavirus type 16 L1 open reading frame results in production of high levels of L1 protein in human epithelial cells. AB - The expression of human papillomavirus type 16 late genes encoding virus capsid proteins L1 and L2 is restricted to terminally differentiated epithelial cells in the superficial layers of the squamous epithelium. We wish to understand the molecular mechanisms that determine the levels of expression of the human papillomavirus type 16 late genes. We have previously shown that the L1 coding region contains inhibitory sequences. Here we extend previous findings to show that the 5' end of the L1 gene contains strong inhibitory sequences but that the 3' end does not. We show that the first 514 nucleotides of the L1 coding region contain multiple inhibitory elements that act independently of one another and that the major inhibitory element is located within the first 129 nucleotides of the L1 gene. Introduction of point mutations in the inhibitory elements in the 5' end of the L1 gene which altered the RNA sequence without affecting the protein sequence specifically inactivated the inhibitory elements and resulted in production of high levels of human papillomavirus type 16 L1 mRNA and protein in human epithelial cells. Furthermore, we show that inhibitory sequences are present in the L1 coding regions of multiple human papillomavirus types, demonstrating that these elements are conserved among the human papillomaviruses, and suggest that they have an important function in the viral life cycle. PMID- 11861840 TI - High-level primary CD8(+) T-cell response to human immunodeficiency virus type 1 gag and env generated by vaccination with recombinant vesicular stomatitis viruses. AB - We investigated the primary cellular immune responses to human immunodeficiency virus type 1 (HIV-1) Env and Gag proteins elicited by recombinant vesicular stomatitis viruses (rVSVs). The primary response to Env peaked 5 to 7 days after intraperitoneal vaccination, at which time 40% of CD8(+) cells were Env tetramer positive and activated (CD62L(Lo)). These freshly isolated cells actively lysed target cells pulsed with the p18-I10 peptide and secreted gamma interferon and tumor necrosis factor alpha after stimulation with the Env p18-I10 peptide. The primary response to Env elicited by rVSVs was sixfold higher than that elicited by recombinant vaccinia viruses (rVVs) at 5 days postvaccination. An intranasal route of vaccination with VSV-Env also elicited a strong primary response to Env. The primary immune response to Gag elicited by rVSV peaked 7 days after vaccination, at which time 3% of CD8(+) cells were Gag tetramer positive and CD62L(Lo) and functional by intracellular cytokine staining. This response was eightfold higher than that elicited by rVV expressing Gag. VSV-GagEnv, which expresses both Gag and Env from a single recombinant, also induced strong cytotoxic T-lymphocyte (CTL) responses to both Env and Gag. Our quantitative analyses illustrate the potency of the VSV vector system in CTL induction. PMID- 11861842 TI - Efficient and selective gene transfer into primary human brain tumors by using single-chain antibody-targeted adenoviral vectors with native tropism abolished. AB - The application of adenoviral vectors in cancer gene therapy is hampered by low receptor expression on tumor cells and high receptor expression on normal epithelial cells. Targeting adenoviral vectors toward tumor cells may improve cancer gene therapy procedures by providing augmented tumor transduction and decreased toxicity to normal tissues. Targeting requires both the complete abolition of native tropism and the addition of a new specific binding ligand onto the viral capsid. Here we accomplished this by using doubly ablated adenoviral vectors, lacking coxsackievirus-adenovirus receptor and alpha(v) integrin binding capacities, together with bispecific single-chain antibodies targeted toward human epidermal growth factor receptor (EGFR) or the epithelial cell adhesion molecule. These vectors efficiently and selectively targeted both alternative receptors on the surface of human cancer cells. Targeted doubly ablated adenoviral vectors were also very efficient and specific with primary human tumor specimens. With primary glioma cell cultures, EGFR targeting augmented the median gene transfer efficiency of doubly ablated adenoviral vectors 123-fold. Moreover, EGFR-targeted doubly ablated vectors were selective for human brain tumors versus the surrounding normal brain tissue. They transduced organotypic glioma and meningioma spheroids with efficiencies similar to those of native adenoviral vectors, while exhibiting greater-than-10-fold reduced background levels on normal brain explants from the same patients. As a result, EGFR-targeted doubly ablated adenoviral vectors had a 5- to 38-fold improved tumor-to-normal brain targeting index compared to native vectors. Hence, single-chain targeted doubly ablated adenoviral vectors are promising tools for cancer gene therapy. They should provide an improved therapeutic index with efficient tumor transduction and effective protection of normal tissue. PMID- 11861843 TI - Analysis of duck hepatitis B virus reverse transcription indicates a common mechanism for the two template switches during plus-strand DNA synthesis. AB - The synthesis of the hepadnavirus relaxed circular DNA genome requires two template switches, primer translocation and circularization, during plus-strand DNA synthesis. Repeated sequences serve as donor and acceptor templates for these template switches, with direct repeat 1 (DR1) and DR2 for primer translocation and 5'r and 3'r for circularization. These donor and acceptor sequences are at, or near, the ends of the minus-strand DNA. Analysis of plus-strand DNA synthesis of duck hepatitis B virus (DHBV) has indicated that there are at least three other cis-acting sequences that make contributions during the synthesis of relaxed circular DNA. These sequences, 5E, M, and 3E, are located near the 5' end, the middle, and the 3' end of minus-strand DNA, respectively. The mechanism by which these sequences contribute to the synthesis of plus-strand DNA was unclear. Our aim was to better understand the mechanism by which 5E and M act. We localized the DHBV 5E element to a short sequence of approximately 30 nucleotides that is 100 nucleotides 3' of DR2 on minus-strand DNA. We found that the new 5E mutants were partially defective for primer translocation/utilization at DR2. They were also invariably defective for circularization. In addition, examination of several new DHBV M variants indicated that they too were defective for primer translocation/utilization and circularization. Thus, this analysis indicated that 5E and M play roles in both primer translocation/utilization and circularization. In conjunction with earlier findings that 3E functions in both template switches, our findings indicate that the processes of primer translocation and circularization share a common underlying mechanism. PMID- 11861844 TI - Analysis of an Autographa californica nucleopolyhedrovirus lef-11 knockout: LEF 11 is essential for viral DNA replication. AB - The Autographa californica nucleopolyhedrovirus (AcMNPV) lef-11 gene was previously identified by transient late expression assays as a gene important for viral late gene expression. The lef-11 gene was not previously identified as necessary for DNA replication in transient origin-dependent plasmid DNA replication assays. To examine the role of lef-11 in the context of the infection cycle, we generated a deletion of the lef-11 gene by recombination in an AcMNPV genome propagated as a BACmid in Escherichia coli. The resulting AcMNPV lef-11 null BACmid (vAc(lef11KO)) was unable to propagate in cell culture, although a "repair" AcMNPV BACmid (vAc(lef11KO-REP)), which was generated by transposition of the lef-11 gene into the polyhedrin locus of the vAc(lef11KO) BACmid, was able to replicate in a manner similar to wild-type or control AcMNPV viruses. Thus, the lef-11 gene is essential for viral replication in Sf9 cells. The vAc(lef11KO) BACmid was examined to determine if the defect in viral replication resulted from a defect in DNA replication or from a defect in late transcription. The lef-11 null BACmid and control BACmids were transfected into Sf9 cells, and viral DNA replication was monitored. The viral DNA genome of the lef-11-null BACmid (vAc(lef11KO)) was not amplified, whereas replication and amplification of the genomes of the repair BACmid (vAc(lef11KO-REP)), wild-type AcMNPV, and a nonpropagating gp64-null control BACmid (vAc(GUSgp64KO)) were readily detected. Northern blot analysis of transcripts from selected early, late, and very late genes showed that late and very late transcription was absent in cells transfected with the lef-11-null BACmid. Thus, in contrast to prior studies using transient replication and late expression assays, studies of a lef-11-null BACmid indicate that LEF-11 is required for viral DNA replication during the infection cycle. PMID- 11861845 TI - Herpes simplex virus selectively induces expression of the CC chemokine RANTES/CCL5 in macrophages through a mechanism dependent on PKR and ICP0. AB - Recruitment of leukocytes is essential for eventual control of virus infections. Macrophages represent a leukocyte population involved in the first line of defense against many infections, including herpes simplex virus (HSV) infection. Through presentation of antigens to T cells and production of cytokines and chemokines, macrophages also constitute an important link between the innate and adaptive immune systems. Here, we have investigated the chemokine expression profile of macrophages after HSV infection and the virus-cell interactions involved. By reverse transcription-PCR and cDNA arrays, we found that HSV type 1 (HSV-1) and HSV-2 induced expression of the CC chemokine RANTES/CCL5 in murine macrophage cell lines and peritoneal cells. The CXC chemokine BCA-1/CXCL13 was also induced in peritoneal cells. Twenty-six other chemokines tested were not affected. Accumulation of RANTES mRNA was detectable after 5 h of infection, was sensitive to UV irradiation of the virus, and was preceded by accumulation of viral immediate-early mRNA and proteins. The viral components responsible for initiation of RANTES expression were examined with virus mutants and RAW 264.7 macrophage-like cells expressing a dominant negative mutant of the double stranded-RNA-activated protein kinase (PKR). The PKR mutant cell line displayed reduced constitutive and HSV-inducible RANTES expression compared to the control cell line. HSV-1 mutants deficient in genes encoding the immediate-early proteins ICP4, ICP22, and ICP27 remained fully capable of inducing RANTES expression in macrophages. By contrast, the ability of an ICP0-deficient HSV-1 mutant to induce RANTES expression was compromised. Thus, HSV selectively induces expression of RANTES in macrophages through a mechanism dependent on cellular PKR and viral ICP0. PMID- 11861846 TI - In vivo interference of Rous sarcoma virus budding by cis expression of a WW domain. AB - For all enveloped viruses, the actual mechanism by which nascent virus particles separate or "pinch off" from the cell surface is largely unknown. In the case of retroviruses, the Gag protein drives the budding process, and the virus release step is directed by the late (L) assembly domain within Gag. A PPPPY motif within the L domain of Rous sarcoma virus (RSV) was previously characterized as being critical for the release of virions and shown to interact in vitro with the WW domain of Yes-associated protein (Yap). To determine whether WW domain-L domain interactions can occur in vivo, we attempted to interfere with the host cell machinery normally recruited to the site of budding by inserting this WW domain in different locations within Gag. At a C-terminal location, the WW(Yap) domain had no effect on budding, suggesting that the intervening I domains (which provide the major region of Gag-Gag interaction) prevent its access to the L domain. When positioned on the other side of the I domains closer to the L domain, the WW(Yap) domain resulted in a dramatic interference of particle release, and confocal microscopy revealed a block to budding on the plasma membrane. Budding was restored by attachment of the heterologous L domain of human immunodeficiency virus type 1 Gag, which does not bind WW(Yap). These findings suggest that cis expression of WW domains can interfere with RSV particle release in vivo via specific, high-affinity interactions at the site of assembly on the plasma membrane, thus preventing host factor accessibility to the L domain and subsequent virus-cell separation. In addition, they suggest that L domain-specific host factors function after Gag proteins begin to interact. PMID- 11861847 TI - Physical association of the K3 protein of gamma-2 herpesvirus 68 with major histocompatibility complex class I molecules with impaired peptide and beta(2) microglobulin assembly. AB - To persist in the presence of an active immune system, viruses encode proteins that decrease expression of major histocompatibility complex class I molecules by using a variety of mechanisms. For example, murine gamma-2 herpesvirus 68 expresses the K3 protein, which causes the rapid turnover of nascent class I molecules. In this report we show that certain mouse class I alleles are more susceptible than others to K3-mediated down regulation. Prior to their rapid degradation, class I molecules in K3-expressing cells exhibit impaired assembly with beta(2)-microglobulin. Furthermore, K3 is detected predominantly in association with class I molecules lacking assembly with high-affinity peptides, including class I molecules associated with the peptide loading complex TAP/tapasin/calreticulin. The detection of K3 with class I assembly intermediates raises the possibility that molecular chaperones involved in class I assembly are involved in K3-mediated class I regulation. PMID- 11861848 TI - Ty3 integrase is required for initiation of reverse transcription. AB - The integrase (IN) encoded by the Saccharomyces cerevisiae retrovirus-like element Ty3 has features found in retrovirus IN proteins including the catalytic triad, an amino-terminal zinc-binding motif, and a nuclear localization sequence. Mutations in the amino- and carboxyl-terminal domains of Ty3 IN cause reduced accumulation of full-length cDNA in the viruslike particles. We show that the reduction in cDNA is accompanied by reduced amounts of early intermediates such as minus-strand, strong-stop DNA. Expression of a capsid (CA)-IN fusion protein (CA-IN) complemented catalytic site and nuclear localization mutants, but not DNA mutants. However, expression of a fusion of CA, reverse transcriptase (RT), and IN (CA-RT-IN) complemented transposition of catalytic site and nuclear localization signal mutants, increased the amount of cDNA in some of the mutants, and complemented transposition of several mutants to low frequencies. Expression of a CA-RT-IN protein with a Ty3 IN catalytic site mutation did not complement transposition of either a Ty3 catalytic site mutant or a nuclear localization mutant but did increase the amount of cDNA in several mutants and complement at least one of the cDNA mutants for transposition. These in vivo data support a model in which independent IN domains can contribute to reverse transcription and integration. We conclude that during reverse transcription, the Ty3 IN domain interacts closely with the polymerase domain and may even constitute a domain within a heterodimeric RT. These studies also suggest that during integration the IN catalytic site and at least portions of the IN carboxyl-terminal domain act in cis. PMID- 11861849 TI - Human immunodeficiency virus type 1-hepatitis C virus coinfection: intraindividual comparison of cellular immune responses against two persistent viruses. AB - Both human immunodeficiency virus type 1 (HIV-1) and hepatitis C virus (HCV) lead to chronic infection in a high percentage of persons, and an expanding epidemic of HIV-1-HCV coinfection has recently been identified. These individuals provide an opportunity for simultaneous assessment of immune responses to two viral infections associated with chronic plasma viremia. In this study we analyzed the breadth and magnitude of the CD8(+)- and CD4(+)-T-lymphocyte responses in 22 individuals infected with both HIV-1 and HCV. A CD8(+)-T-lymphocyte response against HIV-1 was readily detected in all subjects over a broad range of viral loads. In marked contrast, HCV-specific CD8(+)-T-lymphocyte responses were rarely detected, despite viral loads in plasma that were on average 1,000-fold higher. The few HCV-specific responses that were observed were relatively weak and limited in breadth. CD4-proliferative responses against HIV-1 were detected in about half of the coinfected subjects tested, but no proliferative response against any HCV protein was found in these coinfected persons. These data demonstrate a major discordance in immune responses to two persistent RNA viruses. In addition, they show a consistent and profound impairment in cellular immune responses to HCV compared to HIV-1 in HIV-1-HCV-coinfected persons. PMID- 11861850 TI - Antibody-mediated enhancement of human immunodeficiency virus type 1 infectivity is determined by the structure of gp120 and depends on modulation of the gp120 CCR5 interaction. AB - In this study, we characterized the viral determinants of coreceptor usage in relation to susceptibility to antibody-mediated neutralization or enhancement of infectivity by using chimeras of three highly related human immunodeficiency virus type 1 (HIV-1) isolates of different phenotypes. We found that the V3 region was the main determinant of antibody-mediated enhancement and coreceptor specificity but that the overall structure of gp120 was also important for these properties. Constructs susceptible to antibody-mediated enhancement preferentially use CCR5 as a coreceptor, in contrast to constructs that were neutralized or not affected. Using monoclonal antibodies directed against CD4 or CCR5, we were able to show that antibody-mediated enhancement was CD4 dependent. Altogether, our results suggest that the modulation of the interaction of gp120 with CCR5 is the mechanism underlying antibody-mediated enhancement of HIV-1 infectivity. PMID- 11861851 TI - Purification and characterization of oligomeric envelope glycoprotein from a primary R5 subtype B human immunodeficiency virus. AB - Human immunodeficiency virus (HIV) continues to be a major public health problem throughout the world, with high levels of mortality and morbidity associated with AIDS. Considerable efforts to develop an effective vaccine for HIV have been directed towards the generation of cellular, humoral, and mucosal immune responses. A major emphasis of our work has been toward the evaluation of oligomeric (o-gp140) forms of the HIV type 1 (HIV-1) envelope protein for their ability to induce neutralizing antibody responses. We have derived stable CHO cell lines expressing o-gp140 envelope protein from the primary non-syncytium inducing (R5) subtype B strain HIV-1(US4). We have developed an efficient purification strategy to purify oligomers to near homogeneity. Using a combination of three detectors measuring intrinsic viscosity, light scattering, and refractive index, we calculated the molecular mass of the oligomer to be 474 kDa, consistent with either a trimer or a tetramer. The hydrodynamic radius (R(h)) of o-gp140 was determined to be 8.40 nm, compared with 5.07 nm for the monomer. The relatively smaller R(h) of the oligomer suggests that there are indeed differences between the foldings of o-gp140 and gp120. To assess the structural integrity of the purified trimers, we performed a detailed characterization of the glycosylation profile of o-gp140, its ability to bind soluble CD4, and also its ability to bind to a panel of monoclonal antibodies with known epitope specificities for the CD4 binding site, the CD4 inducible site, the V3 loop, and gp41. Immunogenicity studies with rabbits indicated that the purified o-gp140 protein was highly immunogenic and induced high-titer, high avidity antibodies directed predominantly against conformational epitopes. These observations confirm the structural integrity of purified o-gp140 and its potential as a vaccine antigen. PMID- 11861853 TI - Tumor control in a model of bone marrow transplantation and acute liver infiltrating B-cell lymphoma: an unpredicted novel function of cytomegalovirus. AB - Tumor relapse and cytomegalovirus (CMV) infection are major concerns in the therapy of hematopoietic malignancies by bone marrow transplantation (BMT). Little attention so far has been given to a possible pathogenetic interplay between CMV and lymphomas. CMV inhibits stem cell engraftment and hematopoietic reconstitution. Thus, by causing maintenance of bone marrow aplasia and immunodeficiency, CMV could promote tumor relapse. Alternatively, CMV could aid tumor remission. One might think of cytopathogenic infection of tumor cells, induction of apoptosis or inhibitory cytokines, interference with tumor cell extravasation or tumor vascularization, or bystander stimulation of an antitumoral immune response. To approach these questions, the established model of experimental BMT and murine CMV infection was extended by the introduction of liver-infiltrating, highly tumorigenic variant clone E12E of BALB/c-derived B cell lymphoma A20. We document a remarkable retardation of lymphoma progression. First-guess explanations were ruled out: (i) lymphoma cells were not infected; (ii) lymphoma cells located next to infected hepatocytes did not express executioner caspase 3 but were viable and proliferated; (iii) an inhibitory effect of virus on the formation of tumor nodules in the liver became apparent by day 7 after BMT, long before the reconstitution of immune cells; and (iv) recombinant tumor necrosis factor alpha (TNF-alpha) did not substitute for virus; accordingly anti-TNF-alpha did not prevent the inhibition. Notably, while the antitumoral effect required replicative virus, prevention of cytopathogenic infection of the liver by antiviral CD8 T cells did not abolish lymphoma control. These findings are paradigmatic for a novel virus-associated antitumoral mechanism distinct from oncolysis. PMID- 11861852 TI - Solution structure of the viral receptor domain of Tva and its implications in viral entry. AB - Tva is the cellular receptor for subgroup A avian sarcoma and leukosis virus (ASLV-A). The viral receptor function of Tva is determined by a 40-residue, cysteine-rich motif called the LDL-A module. Here we report the solution structure of the LDL-A module of Tva, determined by nuclear magnetic resonance (NMR) spectroscopy. Although the carboxyl terminus of the Tva LDL-A module has a structure similar to those of other reported LDL-A modules, the amino terminus adopts a different conformation. The LDL-A module of Tva does not contain the signature antiparallel beta-sheet observed in other LDL-A modules, and it is more flexible than other reported LDL-A modules. The LDL-A structure of Tva provides mechanistic insights into how a simple viral receptor functions in retrovirus entry. The side chains of H38 and W48 of Tva, which have been identified as viral contact residues by mutational analysis, are solvent exposed, suggesting that they are directly involved in EnvA binding. However, the side chain of L34, another potential viral contact residue identified previously, is buried inside of the module and forms the hydrophobic core with other residues. Thus L34 likely stabilizes the Tva structure but is not a viral interaction determinant. In addition, we propose that the flexible amino-terminal region of Tva plays an important role in determining specificity in the Tva-EnvA interaction. PMID- 11861854 TI - Identification of temperature-sensitive mutations in the phosphoprotein of respiratory syncytial virus that are likely involved in its interaction with the nucleoprotein. AB - The phosphoprotein (P) of human respiratory syncytial virus (RSV) is an essential component of the viral RNA polymerase, along with the large polymerase (L), nucleocapsid (N), and M2-1 proteins. By screening a randomly mutagenized P gene cDNA library, two independent mutations, one with a substitution of glycine at position 172 by serine (G172S) and the other with a substitution of glutamic acid at position 176 by glycine (E176G), were identified to result in the loss of N-P interaction at 37 degrees C in the yeast two-hybrid assay. Both P mutants exhibited greatly reduced activity in supporting the replication and transcription of an RSV minigenome replicon at 37 and 39 degrees C. The G172S and E176G mutations were introduced individually into the RSV A2 (rA2) antigenomic cDNA, and recombinant viruses, rA2-P172 and rA2-P176, were obtained. Both viruses replicate as well as wild-type A2 virus in both Vero and HEp-2 cells at 33 degrees C, but each mutant virus exhibited temperature-sensitive replication in both cell lines. rA2-P176 is more temperature sensitive than rA2-P172. Coimmunoprecipitation of the N protein with each P mutant from virus-infected cells demonstrates that N-P interaction is impaired at 37 degrees C. In addition, the levels of replication of rA2-P172 and rA2-P176 in the lungs of mice and cotton rats were reduced. As is the case with the in vitro assays, rA2-P176 is more restricted in replication in the lower respiratory tract of mice and cotton rats than rA2-P172. During in vitro passage at 37 degrees C, the E176G mutation in rA2-P176 was rapidly changed from glycine to predominantly aspartic acid; mutations to cysteine or serine were also detected. All of the revertants lost the temperature-sensitive phenotype. To analyze the importance of the amino acids in the region from positions 161 to 180 for the P protein function, additional mutations were introduced and their functions were analyzed in vitro. A double mutant containing both G172S and E176G changes in the P gene, substitution of the three charged residues at positions 174 to 176 by alanine, and a deletion of residues from positions 161 to 180 completely abolished the P protein function in the minigenome assay. Thus, the amino acids at positions 172 and 176 and the adjacent charged residues play critical roles in the function of the P protein. PMID- 11861855 TI - Heterologous exchanges of the glycoprotein and the matrix protein in a Novirhabdovirus. AB - Infectious hematopoietic necrosis virus (IHNV) and viral hemorrhagic septicemia virus (VHSV) are two salmonid rhabdoviruses replicating at low temperatures (14 to 20 degrees C). Both viruses belong to the Novirhabdovirus genus, but they are only distantly related and do not cross antigenically. By using a recently developed reverse-genetic system based on IHNV (S. Biacchesi et al., J. Virol. 74:11247-11253, 2000), we investigated the ability to exchange IHNV glycoprotein G with that of VHSV. Thus, the IHNV genome was modified so that the VHSV G gene replaced the complete IHNV G gene. A recombinant virus expressing VHSV G instead of IHNV G, rIHNV-Gvhsv, was generated and was shown to replicate as well as the wild-type rIHNV in cell culture. This study was extended by exchanging IHNV G with that of a fish vesiculovirus able to replicate at high temperatures (up to 28 degrees C), the spring viremia of carp virus (SVCV). rIHNV-Gsvcv was successfully recovered; however, its growth was restricted to 14 to 20 degrees C. These results show the nonspecific sequence requirement for the insertion of heterologous glycoproteins into IHNV virions and also demonstrate that an IHNV protein other than the G protein is responsible for the low-temperature restriction on growth. To determine to what extent the matrix (M) protein interacts with G, a series of chimeric pIHNV constructs in which all or part of the M gene was replaced with the VHSV counterpart was engineered and used to recover the respective recombinant viruses. Despite the very low percentage (38%) of amino acid identity between the IHNV and VHSV matrix proteins, viable chimeric IHNVs, harboring either the matrix protein or both the glycoprotein and the matrix protein from VHSV, were recovered and propagated. Altogether, these data show the extreme flexibility of IHNV to accommodate heterologous structural proteins. PMID- 11861856 TI - Distinct glycoprotein O complexes arise in a post-Golgi compartment of cytomegalovirus-infected cells. AB - Human cytomegalovirus (CMV) glycoproteins H, L, and O (gH, gL, and gO, respectively) form a heterotrimeric disulfide-bonded complex that participates in the fusion of the viral envelope with the host cell membrane. During virus maturation, this complex undergoes a series of intracellular assembly and processing events which are not entirely defined (M. T. Huber and T. Compton, J. Virol. 73:3886-3892, 1999). Here, we demonstrate that gO does not undergo the same posttranslational processing in transfected cells as it does in infected cells. We further determined that gO is modified by O-linked glycosylation and that this terminally processed form is highly enriched in virions. However, during studies of gO processing, novel gO complexes were discovered in CMV virions. The newly identified gO complexes, including gO-gL heterodimers, were not readily detected in CMV-infected cells. Further characterization of the trafficking of gO through the secretory pathway of infected cells localized gH, gL, and gO primarily to the Golgi apparatus and trans-Golgi network, supporting the conclusion that the novel virion-associated gO complexes arise in a post Golgi compartment of infected cells. PMID- 11861857 TI - Distinct roles of adenovirus vector-transduced dendritic cells, myoblasts, and endothelial cells in mediating an immune response against a transgene product. AB - Adenovirus-mediated gene delivery via the intramuscular route efficiently promotes an immune response against the transgene product. In this study, a recombinant adenovirus vector encoding beta-galactosidase (Ad beta Gal) was used to transduce dendritic cells (DC), which are antigen-presenting cells, as well as myoblasts and endothelial cells (EC), neither of which present antigens. C57BL/6 mice received a single intramuscular injection of Ad beta Gal-transduced DC, EC, or myoblasts and were then monitored for anti-beta-galactosidase (anti-beta-Gal) antibody production, induction of gamma interferon-secreting CD8(+) T cells, and protection against melanoma tumor cells expressing beta-Gal. While all transduced cell types were able to elicit an antibody response against the transgene product, the specific isotypes were distinct, with exclusive production of immunoglobulin G2a (IgG2a) antibodies following injection of transduced DC and EC versus equivalent IgG1 and IgG2a responses in mice inoculated with transduced myoblasts. Transduced DC induced a strong ex vivo CD8(+) T-cell response at a level of 50% of the specific response obtained with the Ad beta Gal control. In contrast, this response was 6- to 10-fold-lower in animals injected with transduced myoblasts and EC. Accordingly, only animals injected with transduced DC were protected against a beta-Gal tumor challenge. Thus, in order to induce a strong and protective immune response to an adenovirus-encoded transgene product, it is necessary to transduce cells of dendritic lineage. Importantly, it will be advantageous to block the transduction of DC for adenovirus-based gene therapy strategies. PMID- 11861858 TI - Myxoma virus leukemia-associated protein is responsible for major histocompatibility complex class I and Fas-CD95 down-regulation and defines scrapins, a new group of surface cellular receptor abductor proteins. AB - Down-modulation of major histocompatibility class I (MHC-I) molecules is a viral strategy for survival in the host. Myxoma virus, a member of the Poxviridae family responsible for rabbit myxomatosis, can down-modulate the expression of MHC-I molecules, but the viral factor(s) has not been described. We cloned and characterized a gene coding for an endoplasmic reticulum (ER)-resident protein containing an atypical zinc finger and two transmembrane domains, which we called myxoma virus leukemia-associated protein (MV-LAP). MV-LAP down-regulated surface MHC-I and Fas-CD95 molecules upon transfection; the mechanism probably involves an exacerbation of endocytosis and was lost when the ER retention signal was removed. In addition, the lytic activity of MHC-I-restricted antigen-specific cytolytic T lymphocytes (CTL) against myxoma virus-infected antigen-presenting target cells was significantly reduced, revealing a strong correlation between MHC-I down-regulation by MV-LAP and CTL killing in vitro. In vivo experiments with a knockout virus showed that MV-LAP is a virulence factor, potentially involved in the immunosuppression characteristic of myxomatosis. Data bank analysis revealed that MV-LAP has homologs in herpesviruses and other poxviruses. We propose the name "scrapins" to define a new group of ER-resident surface cellular receptor abductor proteins. The down-regulation of cell surface molecules by scrapins probably helps protect infected cells during viral infections. PMID- 11861859 TI - Evolution of a human immunodeficiency virus type 1 variant with enhanced replication in pig-tailed macaque cells by DNA shuffling. AB - DNA shuffling facilitated the evolution of a human immunodeficiency virus type 1 (HIV-1) variant with enhanced replication in pig-tailed macaque peripheral blood mononuclear cells (pt mPBMC). This variant consists exclusively of HIV-1-derived sequences with the exception of simian immunodeficiency virus (SIV) nef. Sequences spanning the gag-protease-reverse transcriptase (gag-pro-RT) region from several HIV-1 isolates were shuffled and cloned into a parental HIV-1 backbone containing SIV nef. Neither this full-length parent nor any of the unshuffled HIV-1 isolates replicated appreciably or sustainably in pt mPBMC. Upon selection of the shuffled viral libraries by serial passaging in pt mPBMC, a species emerged which replicated at substantially higher levels (50 to 100 ng/ml p24) than any of the HIV-1 parents and most importantly, could be continuously passaged in pt mPBMC. The parental HIV-1 isolates, when selected similarly, became extinct. Analyses of full-length improved proviral clones indicate that multiple recombination events in the shuffled region and adaptive changes in the rest of the genome contributed synergistically to the improved phenotype. This improved variant may prove useful in establishing a pig-tailed macaque model of HIV-1 infection. PMID- 11861861 TI - The gene 10 (UL49.5) product of equine herpesvirus 1 is necessary and sufficient for functional processing of glycoprotein M. AB - The functional cooperation of equine herpesvirus 1 (EHV-1) glycoprotein M (gM) and the gene 10 (UL49.5) product was analyzed. Transient-transfection experiments using gM and UL49.5 expression plasmids as well as RK13 cell lines constitutively expressing UL49.5 (RK49.5) or gM (RKgM) demonstrated that the endo-beta-N acetylglucosaminidase H (endo H)-resistant mature form of gM was detectable only after coexpression of the two proteins. Deletion of the EHV-1 UL49.5-homologous gene 10 in strain KyA resulted in a small-plaque phenotype and up to 190-fold reduced virus titers. The growth defects of the mutant KyA Delta 49.5 virus, which were very similar to those of a gM-negative KyA virus, could be completely compensated for by growth of the mutant virus on RK49.5 cells or by repairing the deletion of gene 10 in the revertant virus KyA Delta 49.5R. Analysis of cells infected with the UL49.5-negative EHV-1 demonstrated that gM was not transported to the trans-Golgi network in the absence of the UL49.5 product. In contrast, gM was efficiently transported and processed to the endo H-resistant mature form in KyA Delta 49.5-infected RK49.5 cells. Furthermore, radioimmunoprecipitation experiments demonstrated that gM maturation was observed only if a 10,000-M(r) protein was coprecipitated with gM in KyA- or KyA Delta 49.5R-infected cells or virions. This protein was absent in cells infected with Ky Delta 49.5 or KyA Delta gM, suggesting that it was the EHV-1 UL49.5 product. Taken together, our results demonstrate that the expression of the EHV-1 UL49.5 product is necessary and sufficient for gM processing and that it is required for efficient virus replication. PMID- 11861860 TI - Infectious and whole inactivated simian immunodeficiency viruses interact similarly with primate dendritic cells (DCs): differential intracellular fate of virions in mature and immature DCs. AB - As potential targets for human immunodeficiency virus type 1 and simian immunodeficiency virus (HIV-1 and SIV), dendritic cells (DCs) likely play a significant role in the onset and spread of infection as well as in the induction of antiviral immunity. Using the SIV-macaque system to study the very early events in DC-virus interactions, we compared chemically inactivated SIV having conformationally and functionally intact envelope glycoproteins (2,2' dithiodipyridine [AT-2] SIV) to infectious and heat-treated SIV. Both human and macaque DCs interact similarly with SIV without detectable effects on DC viability, phenotype, or endocytic function. As assessed by measuring cell associated viral RNA, considerable amounts of virus are captured by the DCs and this is reduced when the virus is heat treated or derived from a strain that expresses low levels of envelope glycoprotein. Immunostaining for SIV proteins and electron microscopy indicated that few intact virus particles are retained at the periphery of the endocytically active, immature DCs. This contrasts with a perinuclear localization of numerous virions in large vesicular compartments deeper within mature DCs (in which macropinocytosis is down-regulated). Both immature and mature DCs are capable of clathrin-coated pit-mediated uptake of SIV, supporting the notion that the receptor-mediated uptake of virus can occur readily in mature DCs. While large numbers of whole viruses were preferentially found in mature DCs, both immature and mature DCs contained similar amounts of viral RNA, suggesting that different uptake/virus entry mechanisms are active in immature and mature DCs. These findings have significant implications for cell-to cell transmission of HIV-1 and SIV and support the use of AT-2 SIV, an authentic but noninfectious form of virus, as a useful tool for studies of processing and presentation of AT-2 SIV antigens by DCs. PMID- 11861862 TI - Alternative fates of keratinocytes transduced by human papillomavirus type 18 E7 during squamous differentiation. AB - The human papillomavirus type 18 (HPV-18) E7 protein promotes S-phase reentry in postmitotic, differentiated keratinocytes in squamous epithelium to facilitate vegetative viral DNA amplification. To examine the nature and fate of the differentiated cells that reenter S phase, organotypic cultures of primary human keratinocytes transduced with HPV-18 E7 were pulse-chase-pulse-labeled with (3)H thymidine ((3)H-TdR) and bromodeoxyuridine (BrdU). The kinetics of the appearance of doubly labeled suprabasal cells demonstrate that E7 expression did not promote prolonged S phase. Rather, there was a considerable lag before a small percentage of the cells reentered another round of S phase. Fluorescence in situ hybridization analysis, indeed, revealed a small fraction of the cells with more than 4n chromosomes in the differentiated strata. Differentiated cells positive for (3)H-TdR, BrdU, or both often had enlarged nuclei or were binucleated. These results suggest that S phase is not followed by cell division, although nuclear division may occur. Interestingly, a significant fraction of differentiated cells that entered S phase subsequently accumulated p27kip1 protein with a kinetics preceding the accumulation of cyclin E. We conclude that E7-transduced, differentiated keratinocytes that enter S phase have two alternative fates: (i) a low percentage of cells undergoes endoreduplication, achieving higher than 4n ploidy, and (ii) a high percentage of cells accumulates the p27kip1, cyclin E, and p21cip1 proteins, resulting in arrest and preventing further S-phase reentry. PMID- 11861863 TI - Viable human cytomegalovirus recombinant virus with an internal deletion of the IE2 86 gene affects late stages of viral replication. AB - Using bacterial artificial chromosome (BAC) technology, we have constructed and characterized a human cytomegalovirus recombinant virus with a mutation in the exon specific for the major immediate-early region 2 (IE2) gene product. The resulting IE2 86-kDa protein (IE2 86) has an internal deletion of amino acids 136 to 290 and is fused at the carboxy terminus to enhanced green fluorescent protein (EGFP). The deletion also removes the promoter and initiator methionine for the p40 form of IE2 and initiator methionine for the p60 form of the protein, and therefore, these late gene products are not produced. The mutant virus IE2 86 Delta SX-EGFP is viable but exhibits altered growth characteristics in tissue culture compared with a full-length wild-type (wt) IE2 86-EGFP virus or a revertant virus. When cells are infected with the mutant virus at a low multiplicity of infection (MOI), there is a marked delay in the production of infectious virus. This is associated with slower cell-to-cell spread of the virus. By immunofluorescence and Western blot analyses, we show that the early steps in the replication of the mutant virus are comparable to those for the wt. Although there is significantly less IE2 protein in the cells infected with the mutant, there is only a modest lag in the initial accumulation of IE1 72 and viral early proteins, and viral DNA replication proceeds normally. The mutation also has only a small effect on the synthesis of the viral major capsid protein. The most notable molecular defect in the mutant virus infection is that the steady-state levels of the pp65 (UL83) and pp28 (UL99) matrix proteins are greatly reduced. In the case of UL83, but not UL99, there is also a corresponding decrease in the amount of mRNA present in cells infected with the mutant virus. PMID- 11861864 TI - SUMO-1 modification of human cytomegalovirus IE1/IE72. AB - Human cytomegalovirus (HCMV) immediate-early protein IE1/IE72 is involved in undermining many cellular processes including cell cycle regulation, apoptosis, nuclear architecture, and gene expression. The multifunctional nature of IE72 suggests that posttranslational modifications may modulate its activities. IE72 is a phosphoprotein and has intrinsic kinase activity (S. Pajovic, E. L. Wong, A. R. Black, and J. C. Azizkhan, Mol. Cell. Biol. 17:6459-6464, 1997). We now demonstrate that IE72 is covalently conjugated to the small ubiquitin-like modifier (SUMO-1). SUMO-1 is an 11.5-kDa protein that is conjugated to multiple proteins and has been reported to exhibit multiple effects, including modulation of protein stability, subcellular localization, and gene expression. A covalently modified protein migrating at approximately 92 kDa, which is stabilized by a SUMO 1 hydrolase inhibitor, is revealed by Western blotting with anti-IE72 of lysates from cells infected with HCMV or cells expressing IE72. SUMO modification of IE72 was confirmed by immunoprecipitation with anti-IE72 and anti-SUMO-1 followed by Western blotting with anti-SUMO-1 and anti-IE72, respectively. Lysine 450 is within a sumoylation consensus site (I,V,L)KXE; changing lysine 450 to arginine by point mutation abolishes SUMO-1 modification of IE72. Inhibition of protein phosphatase 1 and 2A, which increases the phosphorylation of IE72, suppresses the formation of SUMO-1-IE72 conjugates. Both wild-type IE72 and IE72(K450R) localize to nuclear PML oncogenic domains and disrupt them. Studies of protein stability, transactivation, and complementation of IE72-deficient HCMV (CR208) have revealed no significant differences between wild-type IE72 and IE72(K450R). PMID- 11861865 TI - Selectable subgenomic and genome-length dicistronic RNAs derived from an infectious molecular clone of the HCV-N strain of hepatitis C virus replicate efficiently in cultured Huh7 cells. AB - Dicistronic, selectable subgenomic replicons derived from the Con1 strain of hepatitis C virus (HCV) are capable of autonomous replication in cultured Huh7 cells (Lohmann et al., Science 285:110-113, 1999). However, adaptive mutations in the NS3, NS5A, and/or NS5B proteins are required for efficient replication of these RNAs and increase by orders of magnitude the numbers of G418-resistant colonies selected following transfection of Huh7 cells. Here, we demonstrate that a subgenomic replicon (NNeo/3-5B) derived from an infectious molecular clone of a second genotype 1b virus, HCV-N (Beard et al., Hepatology 30:316-324, 1999) is also capable of efficient replication in Huh7 cells. G418-resistant cells selected following transfection with NNeo/3-5B RNA contained abundant NS5A antigen and HCV RNA detectable by Northern analysis. Replicon RNA in one of three clonally isolated cell lines contained no mutations in the NS3-NS5B polyprotein, confirming that adaptive mutations are not required for efficient replication in these cells. However, the deletion of a unique 4-amino-acid insertion that is present within the interferon sensitivity-determining region (ISDR) of the NS5A protein in wild-type HCV-N drastically decreased the number of G418-resistant colonies obtained following transfection of Huh7 cells. This effect could be reversed by inclusion of a previously described Con1 cell culture-adaptive mutation (S2005-->I), confirming that this natural insertion has a controlling role in determining the replication capacity of wild-type HCV-N RNA in Huh7 cells. Additional selectable, dicistronic RNAs encoding NS2-NS5B, E1-NS5B, or the full-length HCV polyprotein were also capable of replication and gave rise to G418-resistant cell clones following transfection of Huh7 cells. We conclude that RNA derived from this documented infectious molecular clone has a unique capacity for replication in Huh7 cells in the absence of additional cell culture-adaptive mutations. PMID- 11861866 TI - Induction of anti-human immunodeficiency virus type 1 (HIV-1) CD8(+) and CD4(+) T cell reactivity by dendritic cells loaded with HIV-1 X4-infected apoptotic cells. AB - T-cell responses to X4 strains of human immunodeficiency virus type 1 (HIV-1) are considered important in controlling progression of HIV-1 infection. We investigated the ability of dendritic cells (DC) and various forms of HIV-1 X4 antigen to induce anti-HIV-1 T-cell responses in autologous peripheral blood mononuclear cells from HIV-1-infected persons. Immature DC loaded with HIV-1 IIIB infected, autologous, apoptotic CD8(-) cells and matured with CD40 ligand induced gamma interferon production in autologous CD8(+) and CD4(+) T cells. In contrast, mature DC loaded with HIV-1 IIIB-infected, necrotic cells or directly infected with cell-free HIV-1 IIIB were poorly immunogenic. Thus, HIV-1-infected cells undergoing apoptosis serve as a rich source of X4 antigen for CD8(+) and CD4(+) T cells by DC. This may be an important mechanism of HIV-1 immunogenicity and provides a strategy for immunotherapy of HIV-1-infected patients on combination antiretroviral therapy. PMID- 11861867 TI - Inhibition of human immunodeficiency virus type 1 activity in vitro by a new self stabilized oligonucleotide with guanosine-thymidine quadruplex motifs. AB - An oligonucleotide with a dimeric hairpin guanosine quadruplex (basket type structure) (dG3T4G3-s), containing phosphorothioate groups, was able to inhibit human immunodeficiency virus type 1 (HIV-1)-induced syncytium formation and virus production (as measured by p24 core antigen expression) in peripheral blood mononuclear cells. This oligonucleotide lacks primary sequence homology with the complementary (antisense) sequences to the HIV-1 genome. Furthermore, this oligonucleotide may have increased nuclease resistance. The activity of this oligonucleotide was increased when the phosphodiester backbone was replaced with a phosphorothioate backbone. In vivo results showed that dG3T4G3-s was capable of blocking the interaction between gp120 and CD4. We also found that dG3T4G3-s specifically inhibits the entry of T-cell line-tropic HIV-1 into cells. This compound is a viable candidate for evaluation as a therapeutic agent against HIV 1 in humans. PMID- 11861868 TI - Expression and self-assembly of norwalk virus capsid protein from venezuelan equine encephalitis virus replicons. AB - The Norwalk virus (NV) capsid protein was expressed using Venezuelan equine encephalitis virus replicon particles (VRP-NV1). VRP-NV1 infection resulted in large numbers of recombinant NV-like particles that were primarily cell associated and were indistinguishable from NV particles produced from baculoviruses. Mutations located in the N-terminal and P1 domains of the NV capsid protein ablated capsid self-assembly in mammalian cells. PMID- 11861869 TI - Construction of a human immunodeficiency virus type 1 (HIV-1) library containing random combinations of amino acid substitutions in the HIV-1 protease due to resistance by protease inhibitors. AB - Human immunodeficiency virus type 1 (HIV-1) heterogeneity contributes to the emergence of drug-resistant virus, escape from host defense systems, and/or conversion of the cellular tropism. To establish an in vitro system to address a heterogeneous virus population, we constructed a library of HIV-1 molecular clones containing a set of random combinations of zero to 11 amino acid substitutions associated with resistance to protease inhibitors by the HIV-1 protease. The complexity (2.1 x 10(5)) of the HIV-1 library pNG-PRL was large enough to cover all of the possible combinations of zero to 11 amino acid substitutions (a total of 4,096 substitutions possible). The T-cell line MT-2 was infected with the HIV-1 library, and resistant viruses were selected after treatment by the protease inhibitor ritonavir (0.03 to 0.30 microM). The viruses that contained three to eight amino acid substitutions could be selected within 2 weeks. These results demonstrate that this HIV-1 library could serve as an alternative in vitro system to analyze the emergence of drug resistance and to evaluate the antiviral activity of novel compounds against multidrug-resistant viruses. PMID- 11861870 TI - Equine infectious anemia virus and the ubiquitin-proteasome system. AB - Some retroviruses contain monoubiquitinated Gag and do not bud efficiently from cells treated with proteasome inhibitors, suggesting an interaction between the ubiquitin-proteasome system and retrovirus assembly. We examined equine infectious anemia virus (EIAV) particles and found that approximately 2% of the p9(Gag) proteins are monoubiquitinated, demonstrating that this Gag protein interacts with an ubiquitinating activity. Different types of proteasome inhibitors were used to determine if proteasome inactivation affects EIAV release from chronically infected cells. Pulse-chase immunoprecipitation and time course immunoblot analyses showed that proteasome inactivation slightly decreased virus release (at most a twofold effect), while it did not affect Gag processing. These results contrast with those obtained with other viruses which are sensitive to these inhibitors. This suggests that, although its Gag is monoubiquitinated, the requirements for EIAV release are somewhat different from those for retroviruses that are sensitive to proteasome inhibitors. PMID- 11861871 TI - Porcine endogenous retrovirus transmission characteristics of an inbred herd of miniature swine. AB - Here we report the identification of inbred miniature swine that failed to produce human-tropic replication-competent porcine endogenous retroviruses (HTRC PERVs), using in vitro coculture assays. When HTRC PERVs were isolated from transmitting animals, all were recombinant viruses, with the receptor-binding domain of PERV-A combining with PERV-C-related sequences. PMID- 11861872 TI - CD28(-/-) mice show defects in cellular and humoral immunity but are able to control infection with murine gammaherpesvirus 68. AB - The role of CD28-dependent costimulatory interactions in the development and maintenance of antiviral immune responses was investigated in a mouse model of gammaherpesvirus infection. CD28(-/-) mice could clear a productive infection with murine gammaherpesvirus 68 (MHV-68), although early lung viral titers were significantly increased. Both CD28(-/-) and CD28(+/+) mice maintained effective long-term control of MHV-68. Gamma interferon responses appeared to develop more slowly in CD28(-/-) mice, while cytotoxic T-cell activity was similar to that in wild-type mice. Splenomegaly developed normally in CD28(-/-) mice, whereas virus specific antibody responses were significantly reduced and aberrant class switching was observed. This work demonstrates that costimulatory interactions involving CD28 are not an absolute requirement for the control of infection with MHV-68. PMID- 11861873 TI - Both R5 and X4 human immunodeficiency virus type 1 variants persist during prolonged therapy with five antiretroviral drugs. AB - A viral reservoir of human immunodeficiency virus type 1 (HIV-1)-infected, resting CD4(+) T cells persists despite suppression of plasma viremia by combination antiretroviral therapy. In a longitudinal analysis of three patients treated with a five-drug regimen, both R5 and X4 HIV-1 variants persisted in the cellular reservoir for up to 3 years. PMID- 11861874 TI - CCR5 and CXCR4 usage by non-clade B human immunodeficiency virus type 1 primary isolates. AB - CCR5 and CXCR4 usage has been studied extensively with a variety of clade B human immunodeficiency virus type 1 (HIV-1) isolates. The determinants of CCR5 coreceptor function are remarkably consistent, with a region critical for fusion and entry located in the CCR5 amino-terminal domain (Nt). In particular, negatively charged amino acids and sulfated tyrosines in the Nt are essential for gp120 binding to CCR5. The same types of residues are important for CXCR4 mediated viral fusion and entry, but they are dispersed throughout the extracellular domains of CXCR4, and their usage is isolate dependent. Here, we report on the determinants of CCR5 and CXCR4 coreceptor function for a panel of non-clade B isolates that are responsible for the majority of new HIV-1 infections worldwide. Consistent with clade B isolates, CXCR4 usage remains isolate dependent and is determined by the overall content of negatively charged and tyrosine residues. Residues in the Nt of CCR5 that are important for fusion and entry of clade B isolates are also important for the entry of all non-clade B HIV-1 isolates that we tested. Surprisingly, we found that in contrast to clade B isolates, a cluster of residues in the second extracellular loop of CCR5 significantly affects fusion and entry of all non-clade B isolates tested. This points to a different mechanism of CCR5 usage by these viruses and may have important implications for the development of HIV-1 inhibitors that target CCR5 coreceptor function. PMID- 11861875 TI - Pseudorabies virus UL36 tegument protein physically interacts with the UL37 protein. AB - The UL36 open reading frame encoding the tegument protein ICP1/2 represents the largest open reading frame in the genome of herpes simplex virus type 1 (HSV-1). Polypeptides homologous to the HSV-1 UL36 protein are present in all subfamilies of HERPESVIRIDAE: We sequenced the UL36 gene of the alphaherpesvirus pseudorabies virus (PrV) and prepared a monospecific polyclonal rabbit antiserum against a bacterial glutathione S-transferase (GST)-UL36 fusion protein for identification of the protein. The antiserum detected a >300-kDa protein in PrV-infected cells and in purified virions. Interestingly, in coprecipitation analyses using radiolabeled infected-cell extracts, the anti-UL36 serum reproducibly coprecipitated the UL37 tegument protein, and antiserum directed against the UL37 protein coprecipitated the UL36 protein. This physical interaction could be verified using yeast two-hybrid analysis which demonstrated that the UL37 protein interacts with a defined region within the amino-terminal part of the UL36 protein. By use of immunogold labeling, capsids which accumulate in the cytoplasm in the absence of the UL37 protein (B. G. Klupp, H. Granzow, E. Mundt, and T. C. Mettenleiter, J. Virol. 75:8927-8936, 2001) as well as wild-type intracytoplasmic and extracellular virions were decorated by the anti-UL36 antiserum, whereas perinuclear primary enveloped virions were not. We postulate that the physical interaction of the UL36 protein, which presumably constitutes the innermost layer of the tegument (Z. Zhou, D. Chen, J. Jakana, F. J. Rixon, and W. Chiu, J. Virol. 73:3210-3218, 1999), with the UL37 protein is an important early step in tegumentation during virion morphogenesis in the cytoplasm. PMID- 11861876 TI - Human B cells immortalized with Epstein-Barr virus upregulate CCR6 and CCR10 and downregulate CXCR4 and CXCR5. AB - Compared to peripheral blood resting B cells, Epstein-Barr virus (EBV) immortalized B cells consistently express CCR6 and CCR10 at high levels and CXCR4 and CXCR5 at low levels. Accordingly, these cells vigorously responded to the ligands of CCR6 and CCR10 but not to those of CXCR4 and CXCR5. In a human EBV negative B-cell line, BJAB, stable expression of EBNA2 upregulated CCR6, while stable expression of EBNA2 as well as LMP1 downregulated CXCR4. On the other hand, upregulation of CCR10 or downregulation of CXCR5 was not induced in BJAB by stable expression of EBNA2 or LMP1. Thus, these changes may be due to a plasmablast-like stage of B-cell differentiation fixed by EBV immortalization. EBV-infected B cells in infectious mononucleosis are known to avoid germinal centers and accumulate under the mucosal surfaces. EBV-associated opportunistic lymphomas also tend to occur in extranodal sites. These preferred sites of in vivo localization are consistent with the unique profile of chemokine receptor expression exhibited by EBV-immortalized B cells. PMID- 11861877 TI - The phosphoprotein (P) binding site resides in the N terminus of the L polymerase subunit of sendai virus. AB - Sendai virus encodes an RNA-dependent RNA polymerase which is composed of the L and P proteins. Site-directed mutagenesis of the N terminus of L has identified amino acids important for binding P. Seven of nine mutants in amino acids 1 to 350 of Sendai L lost the ability to bind to Sendai P, although they were still able to bind the viral C protein. Loss of P binding correlated with the loss of all RNA synthesis activities. Two L mutants gave limited P-L complex formation and limited viral transcription and replication. PMID- 11861878 TI - Efficient human immunodeficiency virus replication requires a fine-tuned level of transcription. AB - Transcription represents a crucial step in the life cycle of human immunodeficiency virus (HIV) and is highly regulated. Here we show that the strength of the viral long terminal repeat (LTR) promoter is optimized for efficient replication. Artificially increasing the rate of LTR-driven transcription was strongly detrimental for viral fitness, and HIV was able to regain replication capacity by selecting for variants with a weaker LTR. Strikingly, the strength of the evolved promoter was equivalent to that of the wild-type LTR. PMID- 11861879 TI - Adaptive evolution of variable region genes encoding an unusual type of immunoglobulin in camelids. AB - A typical immunoglobulin (Ig) molecule is composed of four polypeptide chains: two identical heavy (H) chains and two identical light (L) chains. This tetrameric structure is conserved in almost all jawed vertebrate species. However, it has been discovered that camels and llamas (family: Camelidae) possess a type of dimeric Ig that consists of two H chains only. These H chains do not associate with L chains, and they do not have the first constant region (CH1), which is present in the conventional Ig. In spite of these changes, the dimeric Ig maintains the normal immune function. To understand the evolution of the dimeric Ig, we studied the phylogenetic relationships of the variable region (V(H)H) genes of the dimeric Ig from Camelidae and those (V(H)) of the conventional Ig from mammals. The results showed that the V(H)H genes form a monophyletic cluster within one of the mammalian V(H) groups, group C. We examined the type of selective force in complementarity-determining regions (CDRs) and framework regions (FRs) by comparing the rate of synonymous (dS) and nonsynonymous (dN) substitutions. We found that the results obtained from V(H)H genes were similar to those from V(H) genes in that CDRs showed an excess of dN over dS (indicating positive selection), whereas the reverse was true for FRs (purifying selection). However, when the extent of positive selection or purifying selection was investigated at each codon site, three major differences between V(H)H and V(H) genes were found. That is, very different types of selective force were observed between V(H)H and V(H) genes (1) at the sites that contact the L chain in the conventional Ig, (2) at the sites that interact with the CH1 region in the conventional Ig, and (3) in the H1 loop. Our findings suggest that adaptive evolution has occurred in the functionally important sites of the V(H)H genes to maintain the normal immune function in the dimeric Ig. PMID- 11861880 TI - Evolution of Sp transcription factors. AB - The Sp family of transcription factors binds GC-rich DNA sequences. The ubiquitously expressed Sp1 and Sp3 have been well characterized in mammals. Presented here is the characterization of the only Sp protein expressed in the liver or heart tissue of the teleost fish Fundulus heteroclitus. This protein, fSp3, is most similar to and homologous with mammalian Sp3 proteins. The evolution of the Sp transcription family is described, with Sp1 and Sp3 representing the most recent duplication of the Sp family. Sp4 appears to be the most ancestral member. Sp1, Sp3, and Sp4 form a monophyletic group without Sp2. Sp2 is the least similar of the Sp family and is more similar to the non-Sp transcription factors. These results suggest that Sp2 should not be considered a member of the Sp family. Only two domains (zinc fingers and B domain) share similarity outside the Sp family. The zinc fingers are homologous to other GC binding domains, yet the B domain is homologous to protein-protein interacting domains in the CCAAT-binding/NF-Y transcription factor families. These results suggest that these different domains have different evolutionary histories. PMID- 11861881 TI - Natural selection on the erythrocyte surface. AB - Surface glycoproteins are principal receptors used by pathogens to invade target cells. It has been suggested that mammalian erythrocyte surface glycoproteins function as decoy receptors attracting pathogens to the anucleated erythrocyte and away from their target tissues. Glycophorin A (GYPA) is solely expressed on the erythrocyte surface where it is the most abundant sialoglycoprotein, although its function is unknown. The pathogen decoy hypothesis may be relevant here, as GYPA has been shown in vitro to bind numerous viruses and bacteria, which do not infect erythrocytes. However, it is also a receptor for erythrocyte invasion by the malarial parasite Plasmodium falciparum. Analyses of gypa sequence variation among six higher primates and within a human population show that there is a large excess of replacement (nonsynonymous) substitutions along each primate lineage (particularly on exons 2-4 encoding the extracellular glycosylated domain of GYPA) and a significant excess of polymorphisms in exon 2 (encoding the terminal portion of the extracellular domain) within humans. These two signatures suggest that there has been exceptionally strong positive selection on this receptor driving GYPA divergence during primate evolution and balancing selection maintaining allelic variation within human populations. The pathogen decoy hypothesis alone is adequate to explain both these signatures of between-species and within-species diversifying selection. This has implications for understanding the functions of erythrocyte surface components and their roles in health and disease. PMID- 11861882 TI - Experimental genomic evolution: extensive compensation for loss of DNA ligase activity in a virus. AB - Deletion of the viral ligase gene drastically reduced the fitness of bacteriophage T7 on a ligase-deficient host. Viral evolution recovered much of this fitness during long-term passage, but the final fitness remained below that of the intact virus. Compensatory changes occurred chiefly in genes involved in DNA metabolism: the viral endonuclease, helicase, and DNA polymerase. Two other compensatory changes of unknown function also occurred. Using a method to distinguish compensatory mutations from other beneficial mutations, five additional substitutions from the recovery were shown to enhance adaptation to culture conditions and were not compensatory for the deletion. In contrast to the few previous studies of viral recovery from deletions, the compensatory changes in T7 did not restore the deletion or duplicate major regions of the genome. The ability of this deleted genome to recover much of the lost fitness via mutations in its remaining genes reveals a considerable evolutionary potential to modify the interactions of its elements in maintaining an essential set of functions. PMID- 11861883 TI - Adaptation for horizontal transfer in a homing endonuclease. AB - Selfish genes of no function other than self-propagation are susceptible to degeneration if they become fixed in a population, and regular transfer to new species may be the only means for their long-term persistence. To test this idea we surveyed 24 species of yeast for VDE, a nuclear, intein-associated homing endonuclease gene (HEG) originally discovered in Saccharomyces cerevisiae. Phylogenetic analyses show that horizontal transmission has been a regular occurrence in its evolutionary history. Moreover, VDE appears to be specifically adapted for horizontal transmission. Its 31-bp recognition sequence is an unusually well-conserved region in an unusually well-conserved gene. In addition, the nine nucleotide sites most critical for homing are also unusually well conserved. Such adaptation for horizontal transmission presumably arose as a consequence of selection, both among HEGs at different locations in the genome and among variants at the same location. The frequency of horizontal transmission must therefore be a key feature constraining the distribution and abundance of these genes. PMID- 11861884 TI - Xena, a full-length basal retroelement from tetraodontid fish. AB - Mobile genetic elements are ubiquitous throughout the eukaryote superkingdom. We have sequenced a highly unusual full-length retroelement from the Fugu fish, Takifugu rubripes. This element, which we have named Xena, is similar in structure and sequence to the Penelope retroelement from Drosophila virilis and consists of a single long open reading frame containing a reverse transcriptase domain flanked by identical direct long terminal repeat (LTR) sequences. These LTRs show an organization similar to the terminal repeats already described in the Penelope retrotransposon of Drosophila but are structurally and functionally distinct from the LTRs carried by LTR-retrotransposons. In view of their distinctness, we refer to these repeats as PLTRs (Penelope-LTRs). Whereas the element contains a reverse transcriptase, no other domains or motifs commonly associated with retroelements are present. In the full-length Fugu element, the 5' direct PLTR is preceded by an inverted PLTR fragment. Additional elements, many showing various degrees of deletion, are described from the Fugu genome and from that of the freshwater pufferfish Tetraodon nigroviridis. Many of these additional elements are also preceded by inverted PLTR sequences. Xena-like elements are also described from the genomes of several other organisms. The Penelope-Xena lineage is apparently a basal group within the retrotransposons and therefore represents an evolutionarily important class of retroelement. PMID- 11861885 TI - Extent of gene duplication in the genomes of Drosophila, nematode, and yeast. AB - We conducted a detailed analysis of duplicate genes in three complete genomes: yeast, Drosophila, and Caenorhabditis elegans. For two proteins belonging to the same family we used the criteria: (1) their similarity is > or =I (I = 30% if L > or = 150 a.a. and I = 0.01n + 4.8L(-0.32(1 + exp(-L/1000))) if L < 150 a.a., where n = 6 and L is the length of the alignable region), and (2) the length of the alignable region between the two sequences is > or = 80% of the longer protein. We found it very important to delete isoforms (caused by alternative splicing), same genes with different names, and proteins derived from repetitive elements. We estimated that there were 530, 674, and 1,219 protein families in yeast, Drosophila, and C. elegans, respectively, so, as expected, yeast has the smallest number of duplicate genes. However, for the duplicate pairs with the number of substitutions per synonymous site (K(S)) < 0.01, Drosophila has only seven pairs, whereas yeast has 58 pairs and nematode has 153 pairs. After considering the possible effects of codon usage bias and gene conversion, these numbers became 6, 55, and 147, respectively. Thus, Drosophila appears to have much fewer young duplicate genes than do yeast and nematode. The larger numbers of duplicate pairs with K(S) < 0.01 in yeast and C. elegans were probably largely caused by block duplications. At any rate, it is clear that the genome of Drosophila melanogaster has undergone few gene duplications in the recent past and has much fewer gene families than C. elegans. PMID- 11861886 TI - Phylogenetic relationships, ecological correlates, and molecular evolution within the cavioidea (mammalia, rodentia). AB - A molecular phylogeny of the rodent superfamily Cavioidea was derived using two nuclear sequences (exon #10 of the growth hormone receptor gene and intron #1 of the transthyretin gene) and one mitochondrial gene (12S rRNA). A combined analysis produced a highly derived and well-supported phylogenetic hypothesis that differed from traditional taxonomy primarily in the placement of two taxa. Kerodon, traditionally included within the subfamily Caviinae with guinea pigs and its relatives, is placed sister to the family Hydrochaeridae and closely aligned with the subfamily Dolichotinae. Inclusion of Hydrochaeris within the Caviidae renders the familial classification paraphyletic. Our data further support the taxonomic separation of the families Agoutidae and Dasyproctidae. Both the molecular and traditional morphological interpretations are assessed in testing an ecological constraints hypothesis regarding social behaviors. Whereas traditional taxonomy is consistent with an environmental constraints explanation for social behavior, the molecular data suggest that phylogenetic effects may be a more important factor in the evolution of social behavior in this group. Although lineage-specific rate heterogeneity was identified in all three molecular data sets, no significant support was obtained for the metabolic rate hypothesis. However, both nuclear genes displayed patterns in accordance with the generation time hypothesis. PMID- 11861887 TI - Reconstructing the duplication history of tandemly repeated genes. AB - We present a novel approach to deal with the problem of reconstructing the duplication history of tandemly repeated genes that are supposed to have arisen from unequal recombination. We first describe the mathematical model of evolution by tandem duplication and introduce duplication histories and duplication trees. We then provide a simple recursive algorithm which determines whether or not a given rooted phylogeny can be a duplication history and another algorithm that simulates the unequal recombination process and searches for the best duplication trees according to the maximum parsimony criterion. We use real data sets of human immunoglobulins and T-cell receptors to validate our methods and algorithms. Identity between most parsimonious duplication trees and most parsimonious phylogenies for the same data, combined with the agreement with additional knowledge about the sequences, such as the presence of polymorphisms, shows strong evidence that our reconstruction procedure provides good insights into the duplication histories of these loci. PMID- 11861888 TI - Testing the new animal phylogeny: first use of combined large-subunit and small subunit rRNA gene sequences to classify the protostomes. AB - Although the small-subunit ribosomal RNA (SSU rRNA) gene is widely used in the molecular systematics, few large-subunit (LSU) rRNA gene sequences are known from protostome animals, and the value of the LSU gene for invertebrate systematics has not been explored. The goal of this study is to test whether combined LSU and SSU rRNA gene sequences support the division of protostomes into Ecdysozoa (molting forms) and Lophotrochozoa, as was proposed by Aguinaldo et al. (1997) (Nature 387:489) based on SSU rRNA sequences alone. Nearly complete LSU gene sequences were obtained, and combined LSU + SSU sequences were assembled, for 15 distantly related protostome taxa plus five deuterostome outgroups. When the aligned LSU + SSU sequences were analyzed by tree-building methods (minimum evolution analysis of LogDet-transformed distances, maximum likelihood, and maximum parsimony) and by spectral analysis of LogDet distances, both Ecdysozoa and Lophotrochozoa were indeed strongly supported (e.g., bootstrap values >90%), with higher support than from the SSU sequences alone. Furthermore, with the LogDet-based methods, the LSU + SSU sequences resolved some accepted subgroups within Ecdysozoa and Lophotrochozoa (e.g., the polychaete sequence grouped with the echiuran, and the annelid sequences grouped with the mollusc and lophophorates)-subgroups that SSU-based studies do not reveal. Also, the mollusc sequence grouped with the sequences from lophophorates (brachiopod and phoronid). Like SSU sequences, our LSU + SSU sequences contradict older hypotheses that grouped annelids with arthropods as Articulata, that said flatworms and nematodes were basal bilateralians, and considered lophophorates, nemerteans, and chaetognaths to be deuterostomes. The position of chaetognaths within protostomes remains uncertain: our chaetognath sequence associated with that of an onychophoran, but this was unstable and probably artifactual. Finally, the benefits of combining LSU with SSU sequences for phylogenetic analyses are discussed: LSU adds signal, it can be used at lower taxonomic levels, and its core region is easy to align across distant taxa-but its base frequencies tend to be nonstationary across such taxa. We conclude that molecular systematists should use combined LSU + SSU rRNA genes rather than SSU alone. PMID- 11861889 TI - Molecular clocks in reptiles: life history influences rate of molecular evolution. AB - Life history has been implicated as a determinant of variation in rate of molecular evolution amongst vertebrate species because of a negative correlation between body size and substitution rate for many molecular data sets. Both the generality and the cause of the negative body size trend have been debated, and the validity of key studies has been questioned (particularly concerning the failure to account for phylogenetic bias). In this study, a comparative method has been used to test for an association between a range of life-history variables-such as body size, age at maturity, and clutch size-and DNA substitution rate for three genes (NADH4, cytochrome b, and c-mos). A negative relationship between body size and rate of molecular evolution was found for phylogenetically independent pairs of reptile species spanning turtles, lizards, snakes, crocodile, and tuatara. Although this study was limited by the number of comparisons for which both sequence and life-history data were available, the results suggest that a negative body size trend in rate of molecular evolution may be a general feature of reptile molecular evolution, consistent with similar studies of mammals and birds. This observation has important implications for uncovering the mechanisms of molecular evolution and warns against assuming that related lineages will share the same substitution rate (a local molecular clock) in order to date evolutionary divergences from DNA sequences. PMID- 11861890 TI - Hyaloraphidium curvatum: a linear mitochondrial genome, tRNA editing, and an evolutionary link to lower fungi. AB - We have sequenced the mitochondrial DNA (mtDNA) of Hyaloraphidium curvatum, an organism previously classified as a colorless green alga but now recognized as a lower fungus based on molecular data. The 29.97-kbp mitochondrial chromosome is maintained as a monomeric, linear molecule with identical, inverted repeats (1.43 kbp) at both ends, a rare genome architecture in mitochondria. The genome encodes only 14 known mitochondrial proteins, 7 tRNAs, the large subunit rRNA and small subunit rRNA (SSU rRNA), and 3 ORFs. The SSU rRNA is encoded in two gene pieces that are located 8 kbp apart on the mtDNA. Scrambled and fragmented mitochondrial rRNAs are well known from green algae and alveolate protists but are unprecedented in fungi. Protein genes code for apocytochrome b; cytochrome oxidase 1, 2, and 3, NADH dehydrogenase 1, 2, 3, 4, 4L, 5, and 6, and ATP synthase 6, 8, and 9 subunits, and several of these genes are organized in operon like clusters. The set of seven mitochondrially encoded tRNAs is insufficient to recognize all codons that occur in the mitochondrial protein genes. When taking into account the pronounced codon bias, at least 16 nuclear-encoded tRNAs are assumed to be imported into the mitochondria. Three of the seven predicted mitochondria-encoded tRNA sequences carry mispairings in the first three positions of the acceptor stem. This strongly suggests that these tRNAs are edited by a mechanism similar to the one seen in the fungus Spizellomyces punctatus and the rhizopod amoeba Acanthamoeba castellanii. Our phylogenetic analysis confirms with overwhelming support that H. curvatum is a member of the chytridiomycete fungi, specifically related to the Monoblepharidales. PMID- 11861891 TI - Chorionic gonadotropin has a recent origin within primates and an evolutionary history of selection. AB - Chorionic gonadotropin (CG) is a critical signal in establishing pregnancy in humans and some other primates, but this placentally expressed hormone has not been found in other mammalian orders. The gene for one of its two subunits (CG beta subunit [CGbeta]) arose by duplication from the luteinizing hormone beta subunit gene (LHbeta), present in all mammals tested. In this study, 14 primate and related mammalian species were examined by Southern blotting and DNA sequencing to determine where in mammalian phylogeny the CGbeta gene originated. Bats (order Chiroptera), flying lemur (order Dermoptera), strepsirrhine primates, and tarsiers do not have a CGbeta gene, although they possess one copy of the LHbeta gene. The CGbeta gene first arose in the common ancestor of the anthropoid primates (New World monkeys, Old World monkeys, apes, and humans), after the anthropoids diverged from tarsiers. At least two subsequent duplication events occurred in the catarrhine primates, all of which possess multiple CGbeta copies. The LHbeta-CGbeta family of genes has undergone frequent gene conversion among the catarrhines, as well as periods of strong positive selection in the New World monkeys (platyrrhines). In addition, newly generated DNA sequences from the promoter of the CG alpha subunit gene indicate that platyrrhine monkeys use a different mechanism of alpha gene expression control than that found in catarrhines. PMID- 11861892 TI - Gene density and human nucleotide polymorphism. PMID- 11861893 TI - How many wolbachia supergroups exist? PMID- 11861894 TI - Evidence for selection at multiple allozyme loci across a mussel hybrid zone. PMID- 11861895 TI - Site-specific amino acid replacement matrices from structurally constrained protein evolution simulations. PMID- 11861896 TI - Conserved primers for rapid sequencing of the complete mitochondrial genome from carnivores, applied to three species of bears. PMID- 11861897 TI - Frequent intron loss in the white gene: a cautionary tale for phylogeneticists. PMID- 11861899 TI - Compilation and analysis of group II intron insertions in bacterial genomes: evidence for retroelement behavior. AB - Group II introns are novel genetic elements that have properties of both catalytic RNAs and retroelements. Initially identified in organellar genomes of plants and lower eukaryotes, group II introns are now being discovered in increasing numbers in bacterial genomes. Few of the newly sequenced bacterial introns are correctly identified or annotated by those who sequenced them. Here we have compiled and thoroughly analyzed group II introns and their fragments in bacterial DNA sequences reported to GenBank. Intron distribution in bacterial genomes differs markedly from the distribution in organellar genomes. Bacterial introns are not inserted into conserved genes, are often inserted outside of genes altogether and are frequently fragmented, suggesting a high rate of intron gain and loss. Some introns have multiple natural homing sites while others insert after transcriptional terminators. All bacterial group II introns identified to date encode reverse transcriptase open reading frames and are either active retroelements or derivatives of retroelements. Together, these observations suggest that group II introns in bacteria behave primarily as retroelements rather than as introns, and that the strategy for group II intron survival in bacteria is fundamentally different from intron survival in organelles. PMID- 11861898 TI - A question of size: the eukaryotic proteome and the problems in defining it. AB - We discuss the problems in defining the extent of the proteomes for completely sequenced eukaryotic organisms (i.e. the total number of protein-coding sequences), focusing on yeast, worm, fly and human. (i) Six years after completion of its genome sequence, the true size of the yeast proteome is still not defined. New small genes are still being discovered, and a large number of existing annotations are being called into question, with these questionable ORFs (qORFs) comprising up to one-fifth of the 'current' proteome. We discuss these in the context of an ideal genome-annotation strategy that considers the proteome as a rigorously defined subset of all possible coding sequences ('the orfome'). (ii) Despite the greater apparent complexity of the fly (more cells, more complex physiology, longer lifespan), the nematode worm appears to have more genes. To explain this, we compare the annotated proteomes of worm and fly, relating to both genome-annotation and genome evolution issues. (iii) The unexpectedly small size of the gene complement estimated for the complete human genome provoked much public debate about the nature of biological complexity. However, in the first instance, for the human genome, the relationship between gene number and proteome size is far from simple. We survey the current estimates for the numbers of human genes and, from this, we estimate a range for the size of the human proteome. The determination of this is substantially hampered by the unknown extent of the cohort of pseudogenes ('dead' genes), in combination with the prevalence of alternative splicing. (Further information relating to yeast is available at http://genecensus.org/yeast/orfome) PMID- 11861900 TI - SGS1 is a multicopy suppressor of srs2: functional overlap between DNA helicases. AB - Sgs1 is a member of the RecQ family of DNA helicases, which have been implicated in genomic stability, cancer and ageing. Srs2 is another DNA helicase that shares several phenotypic features with Sgs1 and double sgs1srs2 mutants have a severe synthetic growth phenotype. This suggests that there may be functional overlap between these two DNA helicases. Consistent with this idea, we found the srs2Delta mutant to have a similar genotoxin sensitivity profile and replicative lifespan to the sgs1Delta mutant. In order to directly test if Sgs1 and Srs2 are functionally interchangeable, the ability of high-copy SGS1 and SRS2 plasmids to complement the srs2Delta and sgs1Delta mutants was assessed. We report here that SGS1 is a multicopy suppressor of the methyl methanesulphonate (MMS) and hydroxyurea sensitivity of the srs2Delta mutant, whereas SRS2 overexpression had no complementing ability in the sgs1Delta mutant. Domains of Sgs1 directly required for processing MMS-induced DNA damage, most notably the helicase domain, are also required for complementation of the srs2Delta mutant. Although SGS1 overexpression was unable to rescue the shortened mean replicative lifespan of the srs2Delta mutant, maximum lifespan was significantly increased by multicopy SGS1. We conclude that Sgs1 is able to partially compensate for the loss of Srs2. PMID- 11861901 TI - Identification of HDAC10, a novel class II human histone deacetylase containing a leucine-rich domain. AB - Histone acetylation is important for regulating chromatin structure and gene expression. Three classes of mammalian histone deacetylases have been identified. Among class II, there are five known members, namely HDAC4, HDAC5, HDAC6, HDAC7 and HDAC9. Here we describe the identification and characterization of a novel class II member termed HDAC10. It is a 669 residue polypeptide with a bipartite modular structure consisting of an N-terminal Hda1p-related putative deacetylase domain and a C-terminal leucine-rich domain. HDAC10 is widely expressed in adult human tissues and cultured mammalian cells. It is enriched in the cytoplasm and this enrichment is not sensitive to leptomycin B, a specific inhibitor known to block the nuclear export of other class II members. The leucine-rich domain of HDAC10 is responsible for its cytoplasmic enrichment. Recombinant HDAC10 protein possesses histone deacetylase activity, which is sensitive to trichostatin A, a specific inhibitor for known class I and class II histone deacetylases. When tethered to a promoter, HDAC10 is able to repress transcription. Furthermore, HDAC10 interacts with HDAC3 but not with HDAC4 or HDAC6. These results indicate that HDAC10 is a novel class II histone deacetylase possessing a unique leucine rich domain. PMID- 11861902 TI - Chemical incorporation of 1-methyladenosine into oligonucleotides. AB - The base moiety of 1-N-methyladenosine can be protected with a chloroacetyl group for incorporation of this modified nucleoside into DNA and RNA. Carefully controlled anhydrous conditions are needed for deprotection of the oligonucleotides. PMID- 11861903 TI - The contribution of 2'-hydroxyls to the cleavage activity of the Neurospora VS ribozyme. AB - We have used nucleotide analog interference mapping and site-specific substitution to determine the effect of 2'-deoxynucleotide substitution of each nucleotide in the VS ribozyme on the self-cleavage reaction. A large number of 2' hydroxyls (2'-OHs) that contribute to cleavage activity of the VS ribozyme were found distributed throughout the core of the ribozyme. The locations of these 2' OHs in the context of a recently developed helical orientation model of the VS ribozyme suggest roles in multi-stem junction structure, helix packing, internal loop structure and catalysis. The functional importance of three separate 2'-OHs supports the proposal that three uridine turns contribute to local and long-range tertiary structure formation. A cluster of important 2'-OHs near the loop that is the candidate region for the active site and one very important 2'-OH in the loop that contains the cleavage site confirm the functional importance of these two loops. A cluster of important 2'-OHs lining the minor groove of stem-loop I and helix II suggests that these regions of the backbone may play an important role in positioning helices in the active structure of the ribozyme. PMID- 11861904 TI - Methylation-dependent silencing at the H19 imprinting control region by MeCP2. AB - Methylation of CpG dinucleotides is correlated with transcriptional repression of genes, including imprinted genes. In the case of the imprinted H19 gene, a 2 kb imprinting control region (ICR) is subject to differential methylation, as it is methylated only on the silenced paternal allele. This region has previously been shown to act as a silencer element at the endogenous locus. The proteins that bind at the H19 differentially methylated domain (DMD) and mediate transcriptional silencing have yet to be identified, although a family of proteins containing a methyl-CpG-binding domain (MBD), of which MeCP2 is the best characterised, are obvious candidates. MeCP2 can bind to a single methylated CpG dinucleotide through its MBD and also contains a transcriptional repression domain (TRD). The TRD interacts with Sin3a and histone deacetylases (HDACs) in vivo, forming a repressive complex. Here we show that MeCP2 is recruited to the H19 DMD in vivo and can silence a reporter gene regulated by the H19 DMD in a methylation-dependent manner. This repression can be alleviated by deletion of the TRD from MeCP2 or by inhibition of HDAC activity. These data indicate that transcriptional silencing from the H19 ICR involves recruitment of MeCP2 and presumably an associated protein complex with deacetylase activity. This complex may also be recruited to the ICR in vivo, resulting in a compact, repressive chromatin structure capable of silencing the paternal H19 allele. PMID- 11861905 TI - Glucose-inducible expression of rrg1+ in Schizosaccharomyces pombe: post transcriptional regulation of mRNA stability mediated by the downstream region of the poly(A) site. AB - rrg1+(rapid response to glucose) has been isolated previously as a UV-inducible gene in Schizosaccharomyces pombe, designated as uvi22+. However, it was revealed that the transcript level of this gene was regulated by glucose, not by DNA damaging agents. Glucose depletion led to a rapid decrease in the level of rrg1+ mRNA, by approximately 50% within 30 min. This effect was readily reversed upon re-introduction of glucose within 1 h. High concentrations (4 and 8%) of glucose showed similar effects on increasing the rrg1+ mRNA level compared with 2% glucose, while a low concentration (0.1%) was not effective in raising the rrg1+ mRNA level. In addition, sucrose and fructose could increase rrg1+ mRNA level. Interestingly, the rapid decline in mRNA level seen upon glucose deprivation resulted from precipitous reduction of mRNA half-life. Serial and internal deletions within the 3'-flanking region of rrg1+ revealed that a 210-nt region downstream of the distal poly(A) site was critical for glucose-regulated expression. Moreover, this downstream region participated in 3'-end formation of mRNA. Taken together, this is the first report on glucose-inducible expression regulated post-transcriptionally by control of mRNA stability in S.pombe. PMID- 11861906 TI - Phosphorylation of mammalian translation initiation factor 5 (eIF5) in vitro and in vivo. AB - Eukaryotic translation initiation factor 5 (eIF5) interacts with the 40S initiation complex (40S*eIF3*AUG*Met-tRNA(f)*eIF2*GTP) and, acting as a GTPase activating protein, promotes the hydrolysis of bound GTP. We isolated a protein kinase from rabbit reticulocyte lysates on the basis of its ability to phosphorylate purified bacterially expressed recombinant rat eIF5. Physical, biochemical and antigenic properties of this kinase identify it as casein kinase II (CK II). Mass spectrometric analysis of maximally in vitro phosphorylated eIF5 localized the major phosphorylation sites at Ser-387 and Ser-388 near the C terminus of eIF5. These serine residues are embedded within a cluster of acidic amino acid residues and account for nearly 90% of the total in vitro eIF5 phosphorylation. A minor phosphorylation site at Ser-174 was also observed. Alanine substitution mutagenesis at Ser-387 and Ser-388 of eIF5 abolishes phosphorylation by the purified kinase as well as by crude reticulocyte lysates. The same mutations also abolish phosphorylation of eIF5 when transfected into mammalian cells suggesting that CK II phosphorylates eIF5 at these two serine residues in vivo as well. PMID- 11861907 TI - Interaction generality, a measurement to assess the reliability of a protein protein interaction. AB - Here we introduce the 'interaction generality' measure, a new method for computationally assessing the reliability of protein-protein interactions obtained in biological experiments. This measure is basically the number of proteins involved in a given interaction and also adopts the idea that interactions observed in a complicated interaction network are likely to be true positives. Using a group of yeast protein-protein interactions identified in various biological experiments, we show that interactions with low generalities are more likely to be reproducible in other independent assays. We constructed more reliable networks by eliminating interactions whose generalities were above a particular threshold. The rate of interactions with common cellular roles increased from 63% in the unadjusted estimates to 79% in the refined networks. As a result, the rate of cross-talk between proteins with different cellular roles decreased, enabling very clear predictions of the functions of some unknown proteins. The results suggest that the interaction generality measure will make interaction data more useful in all organisms and may yield insights into the biological roles of the proteins studied. PMID- 11861908 TI - RNase 8, a novel RNase A superfamily ribonuclease expressed uniquely in placenta. AB - We report the identification and characterization of the gene encoding the eighth and final human ribonuclease (RNase) of the highly diversified RNase A superfamily. The RNase 8 gene is linked to seven other RNase A superfamily genes on chromosome 14. It is expressed prominently in the placenta, but is not detected in any other tissues examined. Phylogenetic analysis suggests that RNase 7 is the closest relative of RNase 8 and that the pair likely resulted from a recent gene duplication event in primates. Further analysis reveals that the RNase 8 gene has incorporated non-silent mutations at an elevated rate (1.3 x 10( 9) substitutions/site/year) and that orthologous RNase 8 genes from 6 of 10 primate species examined have been deactivated by frameshifting deletions or point mutations at crucial structural or catalytic residues. The ribonucleolytic activity of recombinant human RNase 8 is among the lowest of members of this superfamily and it exhibits neither antiviral nor antibacterial activities characteristic of some other RNase A ribonucleases. The rapid evolution, species limited deactivation and tissue-specific expression of RNase 8 suggest a unique physiological function and reiterates the evolutionary plasticity of the RNase A superfamily. PMID- 11861909 TI - Directionality of lambda plasmid DNA replication carried out by the heritable replication complex. AB - There are two 'pathways' of replication of lambda plasmids in Escherichia coli. One pathway requires the assembly of a new replication complex before replication and the second pathway is based on the activity of the replication complex inherited by one of two daughter plasmid copies after a preceding replication round. Such a phenomenon was postulated to occur also in other replicons, including Saccharomyces cerevisiae autonomously replicating sequences. Here we investigated directionality of lambda plasmid replication carried out by the heritable and newly assembled replication complexes. Using two-dimensional agarose gel electrophoresis and electron microscopy we demonstrated that in both normal growth conditions and during the relaxed response to amino acid starvation (when only replication carried out by the heritable complex is possible), bidirectionally and undirectionally replicating plasmid molecules occurred in host cells in roughly equal proportions. The results are compatible with the hypothesis that both complexes (heritable and newly assembled) are equivalent. PMID- 11861911 TI - Regularities of context-dependent codon bias in eukaryotic genes. AB - Nucleotides surrounding a codon influence the choice of this particular codon from among the group of possible synonymous codons. The strongest influence on codon usage arises from the nucleotide immediately following the codon and is known as the N1 context. We studied the relative abundance of codons with N1 contexts in genes from four eukaryotes for which the entire genomes have been sequenced: Homo sapiens, Drosophila melanogaster, Caenorhabditis elegans and Arabidopsis thaliana. For all the studied organisms it was found that 90% of the codons have a statistically significant N1 context-dependent codon bias. The relative abundance of each codon with an N1 context was compared with the relative abundance of the same 4mer oligonucleotide in the whole genome. This comparison showed that in about half of all cases the context-dependent codon bias could not be explained by the sequence composition of the genome. Ranking statistics were applied to compare context-dependent codon biases for codons from different synonymous groups. We found regularities in N1 context-dependent codon bias with respect to the codon nucleotide composition. Codons with the same nucleotides in the second and third positions and the same N1 context have a statistically significant correlation of their relative abundances. PMID- 11861910 TI - Protein and drug interactions in the minor groove of DNA. AB - Interactions between proteins, drugs, water and B-DNA minor groove have been analyzed in crystal structures of 60 protein-DNA and 14 drug-DNA complexes. It was found that only purine N3, pyrimidine O2, guanine N2 and deoxyribose O4' are involved in the interactions, and that contacts to N3 and O2 are most frequent and more polar than contacts to O4'. Many protein contacts are mediated by water, possibly to increase the DNA effective surface. Fewer water-mediated contacts are observed in drug complexes. The distributions of ligands around N3 are significantly more compact than around O2, and distributions of water molecules are the most compact. Distributions around O4' are more diffuse than for the base atoms but most distributions still have just one binding site. Ligands bind to N3 and O2 atoms in analogous positions, and simultaneous binding to N3 and N2 in guanines is extremely rare. Contacts with two consecutive nucleotides are much more frequent than base-sugar contacts within one nucleotide. The probable reason for this is the large energy of deformation of hydrogen bonds for the one nucleotide motif. Contacts of Arg, the most frequent amino acid ligand, are stereochemically indistinguishable from the binding of the remaining amino acids except asparagine (Asn) and phenylalanine (Phe). Asn and Phe bind in distinct ways, mostly to a deformed DNA, as in the complexes of TATA-box binding proteins. DNA deformation concentrates on dinucleotide regions with a distinct deformation of the delta and epsilon backbone torsion angles for the Asn and delta, epsilon, zeta and chi for the Phe-contacted regions. PMID- 11861912 TI - Processing of nucleopeptides mimicking the topoisomerase I-DNA covalent complex by tyrosyl-DNA phosphodiesterase. AB - Tyrosyl-DNA phosphodiesterase-1 (Tdp1) is the only known enzyme to remove tyrosine from complexes in which the amino acid is linked to the 3'-end of DNA fragments. Such complexes can be produced following DNA processing by topoisomerase I, and recent studies in yeast have demonstrated the importance of TDP1 for cell survival following topoisomerase I-mediated DNA damage. In the present study, we used synthetic oligodeoxynucleotide-peptide conjugates (nucleopeptides) and recombinant yeast Tdp1 to investigate the molecular determinants for Tdp1 activity. We find that Tdp1 can process nucleopeptides with up to 13 amino acid residues but is poorly active with a 70 kDa fragment of topoisomerase I covalently linked to a suicide DNA substrate. Furthermore, Tdp1 was more effective with nucleopeptides with one to four amino acids than 15 amino acids. Tdp1 was also more effective with nucleopeptides containing 15 nt than with homolog nucleopeptides containing 4 nt. These results suggest that DNA binding contributes to the activity of Tdp1 and that Tdp1 would be most effective after topoisomerase I has been proteolyzed in vivo. PMID- 11861913 TI - The 3'-untranslated regions of cytoskeletal muscle mRNAs inhibit translation by activating the double-stranded RNA-dependent protein kinase PKR. AB - Cytoskeletal proteins are associated with actin in the microfilaments and have a major role in microfilament assembly and function. The expression of some of these proteins has been implicated in cell growth and transformation. Specifically, the 3'-untranslated regions (3'-UTRs) of tropomyosin, troponin and cardiac actin can induce muscle cell differentiation and appear to function as tumor suppressors. These RNA sequences are predicted to fold to form secondary structures with extended stretches of duplex. We show that the 3'-UTRs of the cytoskeletal mRNAs interact with the RNA-binding domain of the RNA-activated protein kinase PKR. Correspondingly, these RNAs activate PKR in vitro and inhibit globin translation in the rabbit reticulocyte lysate translation system. These data are consistent with a mechanism whereby PKR mediates the differentiation- and tumor-related actions of the cytoskeletal 3'-UTR sequences. PMID- 11861914 TI - YY1 activates Msx2 gene independent of bone morphogenetic protein signaling. AB - Msx2 is a homeobox gene expressed in multiple embryonic tissues which functions as a key mediator of numerous developmental processes. YY1 is a bi-functional zinc finger protein that serves as a repressor or activator to a variety of promoters. The role of YY1 during embryogenesis remains unknown. In this study, we report that Msx2 is regulated by YY1 through protein-DNA interactions. During embryogenesis, the expression pattern of YY1 was observed to overlap in part with that of Msx2. Most notably, during first branchial arch and limb development, both YY1 and Msx2 were highly expressed, and their patterns were complementary. To test the hypothesis that YY1 regulates Msx2 gene expression, P19 embryonal cells were used in a number of expression and binding assays. We discovered that, in these cells, YY1 activated endogenous Msx2 gene expression as well as Msx2 promoter-luciferase fusion gene activity. These biological activities were dependent on both the DNA binding and activation domains of YY1. In addition, YY1 bound specifically to three YY1 binding sites on the proximal promoter of Msx2 that accounted for this transactivation. Mutations introduced to these sites reduced the level of YY1 transactivation. As bone morphogenetic protein type 4 (BMP4) regulates Msx2 expression in embryonic tissues and in P19 cells, we further tested whether YY1 is the mediator of this BMP4 activity. BMP4 did not induce the expression of YY1 in early mouse mandibular explants, nor in P19 cells, suggesting that YY1 is not a required mediator of the BMP4 pathway in these tissues at this developmental stage. Taken together, these findings suggest that YY1 functions as an activator for the Msx2 gene, and that this regulation, which is independent of the BMP4 pathway, may be required during early mouse craniofacial and limb morphogenesis. PMID- 11861915 TI - Homologous recombination is essential for RAD51 up-regulation in Saccharomyces cerevisiae following DNA crosslinking damage. AB - We have determined the kinetics of up-regulation of the homologous recombination gene RAD51, one of the genes induced following DNA damage in isogenic haploid DNA repair-deficient mutants of Saccharomyces cerevisiae, using treatment with the DNA crosslinking agent 8-methoxypsoralen. We show that RAD51 is up-regulated concomitantly, although independently, with a shift from the G1 cell cycle phase to G2/M arrest. This up-regulation is absent in homologous recombination repair deficient mutants and increased in mutants deficient in nucleotide excision repair and pol(zeta)-dependent translesion synthesis. We demonstrate that the Rad53-dependent DNA damage signal transduction cascade is active in RAD51 non inducing mutants. However, when independently eliminated, it too abolishes RAD51 up-regulation. We present a model in which RAD51 up-regulation requires two signals: one depending on the Rad53-dependent DNA damage signal transduction cascade and the other on homologous recombination repair. PMID- 11861916 TI - Analysis of similarity within 142 pairs of orthologous intergenic regions of Caenorhabditis elegans and Caenorhabditis briggsae. AB - Patterns of similarity between genomes of related species reflect the distribution of selective constraint within DNA. We analyzed alignments of 142 orthologous intergenic regions of Caenorhabditis elegans and Caenorhabditis briggsae and found a mosaic pattern with regions of high similarity (phylogenetic footprints) interspersed with non-alignable sequences. Footprints cover approximately 20% of intergenic regions, often occur in clumps and are rare within 5' UTRs but common within 3' UTRs. The footprints have a higher ratio of transitions to transversions than expected at random and a higher GC content than the rest of the intergenic region. The number of footprints and the GC content of footprints within an intergenic region are higher when genes are oriented so that their 5' ends form the boundaries of the intergenic region. Overall, the patterns and characteristics identified here, along with other comparative and experimental studies, suggest that many footprints have a regulatory function, although other types of function are also possible. These conclusions may be quite general across eukaryotes, and the characteristics of conserved regulatory elements determined from genomic comparisons can be useful in prediction of regulation sites within individual DNA sequences. PMID- 11861917 TI - Magnesium is required for specific DNA binding of the CREB B-ZIP domain. AB - We have examined binding of the CREB B-ZIP protein domain to double-stranded DNA containing a consensus CRE sequence (5'-TGACGTCA-3'), the related PAR, C/EBP and AP-1 sequences and the unrelated SP1 sequence. DNA binding was assayed in the presence or absence of MgCl2 and/or KCl using two methods: circular dichroism (CD) spectroscopy and electrophoretic mobility shift assay (EMSA). The CD assay allows us to measure equilibrium binding in solution. Thermal denaturation in 150 mM KCl indicates that the CREB B-ZIP domain binds all the DNA sequences, with highest affinity for the CRE site, followed by the PAR (5'-TAACGTTA-3'), C/EBP (5'-TTGCGCAA-3') and AP-1 (5'-TGAGTCA-3') sites. The addition of 10 mM MgCl2 diminished DNA binding to the CRE and PAR DNA sequences and abolished binding to the C/EBP and AP-1 DNA sequences, resulting in more sequence-specific DNA binding. Using 'standard' EMSA conditions (0.25x TBE), CREB bound all the DNA sequences examined. The CREB-CRE complex had an apparent Kd of approximately 300 pM, PAR of approximately 1 nM, C/EBP and AP-1 of approximately 3 nM and SP1 of approximately 30 nM. The addition of 10 mM MgCl2 to the polyacrylamide gel dramatically altered sequence-specific DNA binding. CREB binding affinity for CRE DNA decreased 3-fold, but binding to the other DNA sequences decreased >1000 fold. In the EMSA, addition of 150 mM KCl to the gels had an effect similar to MgCl2. The magnesium concentration needed to prevent non-specific electrostatic interactions between CREB and DNA in solution is in the physiological range and thus changes in magnesium concentration may be a cellular signal that regulates gene expression. PMID- 11861918 TI - Identification and comparative analysis of the chloroplast alpha-subunit gene of DNA-dependent RNA polymerase from seven Euglena species. AB - When the sequence of the Euglena gracilis chloroplast genome was reported in 1993 the alpha-subunit gene (rpoA) of RNA polymerase appeared to be missing, based on a comparison of all putative reading frames to the then known rpoA loci. Since there has been a large increase in known rpoA sequences, the question of a Euglena chloroplast rpoA gene was re-examined. A previously described unknown reading frame of 161 codons was found to be part of an rpoA gene split by a single group III intron. This rpoA gene, which is highly variable from species to species, was then isolated and characterized in five other euglenoid species, Euglena anabaena, Euglena granulata, Euglena myxocylindracea, Euglena stellata and Euglena viridis, and in the Astasia longa plastid genome. All seven Euglena rpoA genes have either one or three group III introns. The rpoA gene products in Euglena spp. appear to be the most variable in this gene family when compared to the rpoA gene in other species of bacteria, algae and plants. Additionally, Euglena rpoA proteins lack a C-terminal domain required for interaction with some regulatory proteins, a feature shared only with some chlorophyte green algae. The E.gracilis rpoA gene is the distal cistron of a multigene cluster that includes genes for carbohydrate biosynthesis, photosynthetic electron transport, an antenna complex and ribosomal proteins. This study provides new insights into the transcription system of euglenoid plastids, the organization of the plastid genome, group III intron evolution and euglenoid phylogeny. PMID- 11861919 TI - Nucleotides of transcription factor binding sites exert interdependent effects on the binding affinities of transcription factors. AB - We can determine the effects of many possible sequence variations in transcription factor binding sites using microarray binding experiments. Analysis of wild-type and mutant Zif268 (Egr1) zinc fingers bound to microarrays containing all possible central 3 bp triplet binding sites indicates that the nucleotides of transcription factor binding sites cannot be treated independently. This indicates that the current practice of characterizing transcription factor binding sites by mutating individual positions of binding sites one base pair at a time does not provide a true picture of the sequence specificity. Similarly, current bioinformatic practices using either just a consensus sequence, or even mononucleotide frequency weight matrices to provide more complete descriptions of transcription factor binding sites, are not accurate in depicting the true binding site specificities, since these methods rely upon the assumption that the nucleotides of binding sites exert independent effects on binding affinity. Our results stress the importance of complete reference tables of all possible binding sites for comparing protein binding preferences for various DNA sequences. We also show results suggesting that microarray binding data using particular subsets of all possible binding sites can be used to extrapolate the relative binding affinities of all possible full length binding sites, given a known binding site for use as a starting sequence for site preference refinement. PMID- 11861920 TI - UV-induced T-->C transition at a TT photoproduct site is dependent on Saccharomyces cerevisiae polymerase eta in vivo. AB - UV-induced reversion of the arg4-17 ochre allele in Saccharomyces cerevisiae is largely dependent on translesion polymerase eta (Rad30p), known to bypass cyclobutane-type TT dimers in an error-free fashion. arg4-17 locus reversion was predominantly due to T-->C transition of T127, the 3' T of a TT photoproduct site. This event was at least 20-fold reduced in a rad30 deletion mutant, irrespective of the status of nucleotide excision repair. These data correlate with known properties of 6-4 TT photoproducts and in vitro characteristics of polymerase eta and suggest that polymerase eta plays an important in vivo role in inserting G opposite the 3' T of 6-4 TT photoproducts at this site. Alternatively, an unprecedented error-prone processing of cyclobutane-type photoproducts at this site by polymerase eta must be assumed as the critical mechanism. Whereas photoreactivation results indeed hint at the latter possibility, a possible regulatory influence of reducing the overall UV damage load on the bypass probability of non-cyclobutane-type pyrimidine dimer photoproducts should not be dismissed. PMID- 11861921 TI - BALSA: Bayesian algorithm for local sequence alignment. AB - The Smith-Waterman algorithm yields a single alignment, which, albeit optimal, can be strongly affected by the choice of the scoring matrix and the gap penalties. Additionally, the scores obtained are dependent upon the lengths of the aligned sequences, requiring a post-analysis conversion. To overcome some of these shortcomings, we developed a Bayesian algorithm for local sequence alignment (BALSA), that takes into account the uncertainty associated with all unknown variables by incorporating in its forward sums a series of scoring matrices, gap parameters and all possible alignments. The algorithm can return both the joint and the marginal optimal alignments, samples of alignments drawn from the posterior distribution and the posterior probabilities of gap penalties and scoring matrices. Furthermore, it automatically adjusts for variations in sequence lengths. BALSA was compared with SSEARCH, to date the best performing dynamic programming algorithm in the detection of structural neighbors. Using the SCOP databases PDB40D-B and PDB90D-B, BALSA detected 19.8 and 41.3% of remote homologs whereas SSEARCH detected 18.4 and 38% at an error rate of 1% errors per query over the databases, respectively. PMID- 11861922 TI - Mutations in domain V of the 23S ribosomal RNA of Bacillus subtilis that inactivate its protein folding property in vitro. AB - The active site of a protein folding reaction is in domain V of the 23S rRNA in the bacterial ribosome and its homologs in other organisms. This domain has long been known as the peptidyl transferase center. Domain V of Bacillus subtilis is split into two segments, the more conserved large peptidyl transferase loop (RNA1) and the rest (RNA2). These two segments together act as a protein folding modulator as well as the complete domain V RNA. A number of site-directed mutations were introduced in RNA1 and RNA2 of B.subtilis, taking clues from reports of these sites being involved in various steps of protein synthesis. For example, sites like G2505, U2506, U2584 and U2585 in Escherichia coli RNA1 region are protected by deacylated tRNA at high Mg2+ concentration and A2602 is protected by amino acyl tRNA when the P site remains occupied already. Mutations A2058G and A2059G in the RNA1 region render the ribosome Ery(r )in E.coli and Lnc(r )in tobacco chloroplast. Sites in P loop G2252 and G2253 in E.coli are protected against modification by the CCA end of the P site bound tRNA. Mutations were introduced in corresponding nucleotides in B.subtilis RNA1 and RNA2 of domain V. The mutants were tested for refolding using unfolded protein binding assays with unfolded carbonic anhydrase. In the protein folding assay, the mutants showed partial to complete loss of this activity. In the filter binding assay for the RNA-refolding protein complex, the mutants showed an extent of protein binding that agreed well with their protein folding activity. PMID- 11861923 TI - An improved helper phage system for efficient isolation of specific antibody molecules in phage display. AB - Phage display technology has been applied in many fields of biological and medical sciences to study molecular interactions and especially in the generation of monoclonal antibodies of human origin. However, extremely low display level of antibody molecules on the surface of phage is an intrinsic problem of a phagemid based display system resulting in low success rate of isolating specific binding molecules. We show here that display of single-chain antibody fragment (scFv) generated with pIGT3 phagemid can be increased dramatically by using a genetically modified Ex-phage. Ex-phage has a mutant pIII gene that produces a functional wild-type pIII in suppressing Escherichia coli strains but does not make any pIII in non-suppressing E.coli strains. Packaging phagemids encoding antibody-pIII fusion in F+ non-suppressing E.coli strains with Ex-phage enhanced the display level of antibody fragments on the surfaces of recombinant phage particles resulting in an increase of antigen-binding reactivity >100-fold compared to packaging with M13KO7 helper phage. Thus, the Ex-phage and pIGT3 phagemid vector provides a system for the efficient enrichment of specific binding antibodies from a phage display library and, thereby, increases the chance of obtaining more diverse antibodies specific for target antigens. PMID- 11861924 TI - Single nucleotide polymorphism detection by combinatorial fluorescence energy transfer tags and biotinylated dideoxynucleotides. AB - Combinatorial fluorescence energy transfer (CFET) tags, constructed by exploiting energy transfer and combinatorial synthesis, allow multiple biological targets to be analyzed simultaneously. We here describe a multiplex single nucleotide polymorphism (SNP) assay based on single base extension (SBE) using CFET tags and biotinylated dideoxynucleotides (biotin-ddNTPs). A library of CFET-labeled oligonucleotide primers was mixed with biotin-ddNTPs, DNA polymerase and the DNA templates containing the SNPs in a single tube. The nucleotide at the 3'-end of each CFET-labeled oligonucleotide primer was complementary to a particular SNP in the template. Only the CFET-labeled primer that is fully complementary to the DNA template was extended by DNA polymerase with a biotin-ddNTP. We isolated the DNA extension fragments that carry a biotin at the 3'-end by capture with streptavidin-coated magnetic beads, while the unextended primers were eliminated. The biotinylated fluorescent DNA fragments were subsequently analyzed in a multicolor fluorescence electrophoresis system. The distinct fluorescence signature and electrophoretic mobility of each DNA extension product in the electropherogram coded the SNPs without the use of a sizing standard. We simultaneously distinguished six nucleotide variations in synthetic DNA templates and a PCR product from the retinoblastoma tumor suppressor gene. The use of CFET labeled primers and biotin-ddNTPs coupled with the specificity of DNA polymerase in SBE offered a multiplex method for detecting SNPs. PMID- 11861925 TI - A novel medium throughput quantitative competitive PCR technology to simultaneously measure mRNA levels from multiple genes. AB - There is a great demand for technologies to simultaneously measure mRNA levels from multiple genes. Here we report a new quantitative competitive PCR technology and demonstrate simultaneous quantification of mRNA from multiple genes. First, a sequential 2-fold dilution series containing equal amounts of gene-specific standard DNAs for 10-12 genes is prepared. Second, the serially diluted standard DNAs are individually added to equal amounts of tissue-derived cDNA and amplified with gene-specific primers for 19-26 PCR cycles. Each gene/standard DNA pair is amplified individually. All amplified DNA products (n = 80) are resolved by one microplate array diagonal gel electrophoresis using 5% polyacrylamide. Changes in mRNA levels of approximately 15% can be detected by this technology. The mRNA levels from 10-12 genes were simultaneously quantified. mRNA levels were compared in RNA samples from rat liver, kidney and skeletal muscle. This quick, specific, sensitive, reproducible and yet inexpensive technique is ideal for simultaneously studying co-ordinate changes in mRNA levels from multiple genes. PMID- 11861926 TI - Tumour class prediction and discovery by microarray-based DNA methylation analysis. AB - Aberrant DNA methylation of CpG sites is among the earliest and most frequent alterations in cancer. Several studies suggest that aberrant methylation occurs in a tumour type-specific manner. However, large-scale analysis of candidate genes has so far been hampered by the lack of high throughput assays for methylation detection. We have developed the first microarray-based technique which allows genome-wide assessment of selected CpG dinucleotides as well as quantification of methylation at each site. Several hundred CpG sites were screened in 76 samples from four different human tumour types and corresponding healthy controls. Discriminative CpG dinucleotides were identified for different tissue type distinctions and used to predict the tumour class of as yet unknown samples with high accuracy using machine learning techniques. Some CpG dinucleotides correlate with progression to malignancy, whereas others are methylated in a tissue-specific manner independent of malignancy. Our results demonstrate that genome-wide analysis of methylation patterns combined with supervised and unsupervised machine learning techniques constitute a powerful novel tool to classify human cancers. PMID- 11861927 TI - Facile method for automated genotyping of single nucleotide polymorphisms by mass spectrometry. AB - In the future, analysis of single nucleotide polymorphisms (SNPs) should become a powerful tool for many genetic applications in areas such as association studies, pharmacogenetics and traceability in the agro-alimentary sector. A number of technologies have been developed for high-throughput genotyping of SNPs. Here we present the simplified GOOD assay for SNP genotyping by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI). The simplified GOOD assay is a single-tube, purification-free, three-step procedure consisting of PCR, primer extension and phosphodiesterase II digestion followed by mass spectrometric analysis. Due to the application of charge-tag technology, no sample purification is required prior to the otherwise very impurity-sensitive MALDI analysis. The use of methylphosphonate containing primers and ddNTPs or alpha-S-ddNTPs together with a novel DNA polymerase derived from Thermotoga maritima for primer extension allow the fluent preparation of negatively charge tagged, allele-specific products. A key feature of this polymerase is its preference for ddNTPs and alpha-S-ddNTPs over dNTPs. The simplified GOOD assay was run with automatic liquid handling at the lowest manageable volumes, automatic data acquisition and interpretation. We applied this novel procedure to genotyping SNPs of candidate genes for hypertension and cardiovascular disease. PMID- 11861928 TI - Arachidonate metabolism in neonates. PMID- 11861929 TI - Efficacy of dietary arachidonic acid provided as triglyceride or phospholipid as substrates for brain arachidonic acid accretion in baboon neonates. AB - Arachidonic acid (AA) is a long-chain polyunsaturate (LCP) present in human breast milk as both triglyceride (TG) and as phospholipid (PL). There has been little attention to the metabolic consequences of lipid form of AA in infant formulas. Our objective was to investigate the efficacy of dietary TG and PL as carriers of AA for accretion in the brain and associated organs of term baboon neonates consuming a formula with LCP composition typical of human milk. TG and phosphatidylcholine (PC) with [U-(13)C]-AA in the sn-2 position and with unlabeled 16:0 in the remaining positions (TG-AA* or PL-AA*, respectively) were used as tracers to study the tissue AA* incorporation. Baboon neonates received a single oral dose of either TG-AA* (n = 3) or PL-AA* (n = 4) at 18-19 d of life. Tissues were obtained 10 d later (28-29 d of life) and isotopic enrichment was measured. In the brain, 4.5% of the PL-AA* dose and 2.1% of the TG-AA* dose were recovered as brain AA*, respectively, indicating that PL was about 2.1-fold more effective than TG as a substrate for brain AA accretion. Preferential incorporation of PL-derived AA* over TG source of AA* was also observed in the liver, lung, plasma, and erythrocytes. Because of the quantitative predominance of TG-AA in formula, total brain AA accretion, expressed as absolute weight, was 5.0-fold greater from TG-AA than from PL-AA. We estimate that about half of postnatal brain AA accretion is derived from dietary preformed AA in term baboon neonates consuming a formula with lipid composition similar to that of human milk. PMID- 11861930 TI - Fatty acid formula supplementation and neuromotor development in rhesus monkey neonates. AB - Docosahexaenoic acid (DHA) is an omega-3 fatty acid that is highly concentrated in CNS tissues. Although breast milk contains the fatty acids DHA and arachidonic acid, infant formulas marketed in North America do not contain these nutrients. The potential deleterious effects of rearing infants with formulas devoid of these nutrients was assessed by comparing nursery-reared rhesus macaque infants (Macaca mulatta) fed standard formula with infants fed standard formula supplemented with physiologically relevant concentrations of DHA (1.0%) and arachidonic acid (1.0%). Neurobehavioral assessments were conducted on d 7, 14, 21, and 30 of life using blinded raters. The 30-min assessment consisted of 45 test items measuring orienting, temperament, reflex capabilities, and motor skills. Plasma concentrations of DHA in standard formula-fed infants were significantly lower than those fed supplemented formula or mother-raised (breast fed) infants; however, infants fed the supplemented formula exhibited higher arachidonic acid levels than either mother-reared infants or infants fed standard formula. Infant monkeys fed the supplemented formula exhibited stronger orienting and motor skills than infants fed the standard formula, with the differences most pronounced during d 7 and 14. This pattern suggests an earlier maturation of specific visual and motor abilities in the supplemented infants. Supplementation did not affect measures of activity or state control, indicating no effect on temperament. These data support the assertion that preformed DHA and arachidonic acid in infant formulas are required for optimal development. PMID- 11861931 TI - Dietary long-chain polyunsaturated fatty acids minimize dexamethasone-induced reductions in arachidonic acid status but not bone mineral content in piglets. AB - The primary objective of this study was to determine whether exogenous arachidonic acid (AA) in a supplemented formula substitute for piglets or sow milk would attenuate reductions in AA status, growth, and bone mineral content (BMC) as a result of exogenous glucocorticoid excess using dexamethasone (DEX). A secondary objective was to confirm a positive effect of exogenous AA on growth and BMC of piglets fed formula and not treated with DEX as well as to determine whether the elevation in BMC was related to greater production of prostaglandin E(2) in bone. Forty-eight 5-d-old male piglets were randomized to be suckled or receive either a standard formula or the same formula, but containing AA (0.5% wt/wt total fat) for 15 d in addition to either treatment with DEX or placebo. Piglets treated with DEX grew slower and had lower BMC of whole body, lumbar spine, and femur in addition to lower proportions of AA, but those fed standard formula had the greatest reductions. Piglets in the supplemented group weighed more than piglets fed standard formula or suckled in both the DEX and placebo groups. Suckled piglets had the highest BMC of whole body and femur compared with standard formula, and the supplemented group was intermediate for whole body but similar to suckled pigs for femur. Release of prostaglandin E(2) was elevated only with supplementation of AA. These data indicate that supplemental AA is associated with elevated whole body and femur BMC but that BMC is not enhanced during glucocorticoid treatment. PMID- 11861932 TI - Cecal colonization and systemic spread of Candida albicans in mice treated with antibiotics and dexamethasone. AB - Infections with Candida albicans have become a significant problem among very low birth weight infants in the neonatal intensive care unit. Risk factors are multiple and include administration of antibiotics and glucocorticoids, such as dexamethasone. Experiments were designed to study the combined effect of oral broad-spectrum antibiotics and parenteral dexamethasone on cecal colonization and extraintestinal dissemination of C. albicans in separate groups of mice that were orally inoculated with one of four C. albicans strains that were either wild-type INT1/INT1 or had one or more disruptions of the INT1 gene. Intestinal colonization was monitored by quantitative culture of the mouse cecum, and extraintestinal invasion was monitored by quantitative culture of the draining mesenteric lymph nodes and kidneys. At sacrifice, the average numbers of cecal C. albicans differed from 7.7 log(10)/g to 6.7 log(10)/g (p < 0.01) in mice orally inoculated with C. albicans containing two functional copies of INT1 and no functional copies of INT1, respectively. The incidence of extraintestinal dissemination to mesenteric lymph nodes and kidneys correspondingly varied from 57 to 13% (p < 0.01) and 83 to 4% (p < 0.01) in mice inoculated with these two C. albicans strains. Mice orally inoculated with C. albicans containing one functional copy of INT1 had intermediate levels of cecal colonization and extraintestinal dissemination. Thus, cecal colonization and extraintestinal dissemination of C. albicans was facilitated in antibiotic-treated mice given dexamethasone. In addition, the presence of two functional copies of the INT1 gene was associated with the greatest levels of cecal colonization and extraintestinal dissemination of C. albicans. PMID- 11861933 TI - Neutrophil CD64 expression: a sensitive diagnostic marker for late-onset nosocomial infection in very low birthweight infants. AB - This study aims to evaluate the diagnostic utilities of four leukocyte surface antigens-two lymphocyte antigens (CD25 and CD45RO) and two neutrophil antigens (CD11b and CD64)-for identification of late-onset nosocomial bacterial infection in preterm, very low birthweight infants, and to define the optimal cutoff value for each marker so that it may act as a reference with which future studies can be compared. Very low birthweight infants in whom infection was suspected when they were >72 h of age were eligible for the study. A full sepsis screen was performed in each episode. IL-6, C-reactive protein, and leukocyte surface antigens (CD25, CD45RO, CD11b, and CD64) were measured at 0 (at the time of sepsis evaluation), 24, and 48 h by standard biochemical methods and quantitative flow cytometric analysis. The diagnostic utilities including sensitivity, specificity, and positive and negative predictive values of each marker and combination of markers for predicting late-onset neonatal infection were determined. One hundred twenty-seven episodes of suspected clinical sepsis were investigated in 80 infants. Thirty-seven episodes were proven infection. The calculated optimal cutoff values for CD25, CD45RO, CD11b, and CD64 were 3,100, 2,900, 10,450, and 4,000 phycoerythrin-molecules bound per cell, respectively. An interim analysis of data after 68 episodes suggested that CD25 and CD45RO were poor predictors of neonatal infection with sensitivity or specificity <75% during a single measurement. Thus, these two markers were excluded from further investigation. In the final analysis, CD64 has the highest sensitivity (95-97%) and negative predictive value (97-99%) at 0 and 24 h after the onset. The addition of IL-6 or C-reactive protein (0 h) to CD64 (24 h) further enhanced the sensitivity and negative predictive value to 100%, and has the specificity and positive predictive value exceeding 88% and 80%, respectively. Neutrophil CD64 expression is a very sensitive marker for diagnosing late-onset nosocomial infection in very low birthweight infants. If further validated, the use of CD64 as an infection marker should allow early discontinuation of antibiotic treatment at 24 h without waiting for the definitive microbiologic culture results. The quantitative flow cytometric analysis applied in this study could be developed into a routine clinical test with high comparability and reproducibility across different laboratories. PMID- 11861934 TI - Cytokine expression of cord and adult blood mononuclear cells in response to Streptococcus agalactiae. AB - Neonatal bacterial sepsis is often characterized by a fulminant clinical course and highly elevated plasma levels of proinflammatory cytokines. To evaluate in vitro activation of the neonatal immune system by specific infectious stimuli, cord blood cells from healthy neonates were examined for expression of tumor necrosis factor-alpha (TNF-alpha), IL-1beta, IL-6, and IL-8 in response to Streptococcus agalactiae (GBS), lipopolysaccharide (LPS), and lipoteichoic acid (LTA). Cytokine-expression was compared in mononuclear cells from cord and adult peripheral blood. TNF-alpha and IL-6 levels in the supernatant of cord blood cell cultures were significantly higher after stimulation with heat-killed GBS (10(7)/mL) than with LPS (2 microg/mL) or LTA (2 microg/mL) (TNF-alpha: 2215 versus 267.5 versus 40 pg/mL, p = 0.001; IL-6: 9667 versus 4909 versus 919 pg/mL, p = 0.006). mRNA expression of TNF-alpha, IL-1beta, IL-6, and IL-8 was equally pronounced after stimulation with either GBS, LPS, or LTA in cord or adult blood cells at various times. A MAb directed against the monocyte receptor molecule CD14 did not inhibit the release of cytokines in cord blood mononuclear cells after stimulation with GBS. In summary, activation of cord blood cells by infectious stimuli is comparable to the adult immune response in terms of expression of proinflammatory cytokines. GBS in particular proves to be a potent activator of the neonatal immune system when compared with LPS and LTA. CD14 seems not to be a crucial molecule for activation of cord blood cells by GBS. PMID- 11861935 TI - Hemodynamic disturbances in premature infants born after chorioamnionitis: association with cord blood cytokine concentrations. AB - Chorioamnionitis and elevated cord blood inflammatory cytokine concentrations are associated with detectable disturbances of systemic and cerebral hemodynamics in premature newborns. Fifty-five infants (25-31 wk gestation) were enrolled. Chorioamnionitis was defined by placental histology. IL-6, IL-1beta, and tumor necrosis factor-alpha were quantified by ELISA. Blood pressure, heart rate, cardiac output, stroke volume, fractional shortening, and middle cerebral artery blood flow velocities were measured at 3 +/- 1 h after birth. Chorioamnionitis was evident in 22 placentas and was associated with increased IL-6 (p < 0.001), IL-1beta (p = 0.035), and heart rate (p = 0.027); and with decreased mean and diastolic blood pressure (p = 0.026 and p = 0.019, respectively). IL-6 concentration correlated inversely with systolic, mean, and diastolic blood pressures. Right ventricular cardiac output was elevated (p = 0.028) in infants with fetal vessel inflammation. Maternal temperature >or=38.0 degrees C and newborn immature-to-total white blood cell ratio >or=0.4 were associated with significant decreases in left ventricular fractional shortening (p = 0.001 and p = 0.005, respectively). Neither chorioamnionitis nor elevated cytokine concentrations were associated with changes in middle cerebral artery Doppler blood flow velocities. Chorioamnionitis and elevated cord blood IL-6 concentrations are associated with decreased blood pressure in premature newborns. Inflammation of the fetal vessels and nonspecific indicators of infection are associated with disturbances in cardiac function. Infants with chorioamnionitis and elevated cytokine concentrations do not manifest changes in cerebral Doppler indices within the first few postnatal hours. We speculate that cytokine-associated systemic hemodynamic disturbances in premature infants born after chorioamnionitis predispose such infants to perinatal brain injury. PMID- 11861936 TI - Enhanced interleukin-6 and interleukin-8 synthesis in term and preterm infants. AB - There is growing evidence that sepsis-related complications in neonates are crucially mediated by the action of proinflammatory cytokines. It has previously been demonstrated that elevated IL-6 and IL-8 levels can predict brain damage and chronic lung disease in preterm infants. However, it is the current view that neonates have a reduced capability to produce proinflammatory cytokines. To clarify this issue, we analyzed the inflammatory response in term and preterm infants directly at the single cell level by flow cytometry. Endotoxin challenge was performed under defined conditions on monocytes obtained from 50 healthy adults and 119 neonates, which consist of 45 term infants, 63 preterm infants (26.1-36.7 wk of gestational age), and 11 preterm infants with proven infection (24.6-29.9 wk). Our results challenge the existing view of an immature inflammatory response by demonstrating that term infants and preterm infants display a higher percentage of IL-6- and IL-8-positive cells than adults. After preincubation with dexamethasone the number of cytokine-positive cells decreased in all groups, but the number of IL-8-positive cells remained higher in term and preterm infants >32 wk compared with adults. These observations demonstrate not only a well-developed but also an enhanced inflammatory response in term and preterm infants. Under consideration of several detrimental effects of IL-6 and IL-8, our data may have major implications on the pathophysiology of inflammatory triggered neonatal diseases. PMID- 11861937 TI - IGF-related proteins at birth in a case of antenatally diagnosed Silver-Russell syndrome. AB - We report here a case with severe intrauterine growth restriction from the first trimester and clinical features of Silver-Russell syndrome including microcephaly, low-set ear, atrial septum defect, ventricular septum defect, diaphragmatic relaxation, and rocker bottom feet. Silver-Russell syndrome is thought to result from deletion of the distal long arm of chromosome 15 on which the IGF-I receptor (IGF-IR) gene is located. We measured both the maternal and cord blood levels of GH, IGF-I, and IGF-binding protein and performed an immunohistochemical study of IGF-IR in the placenta to investigate whether these IGF-related proteins were affected in this patient. The hormonal level of these proteins did not significantly differ from normal neonates, and immunohistochemical analysis suggested IGF-IR protein levels in the placenta were comparable to normal term neonates. These results support the hypothesis that growth insufficiency could occur in patients with monosomy of the distal long arm of chromosome 15 and suggest a critical threshold for IGF-related fetal growth in early pregnancy. PMID- 11861938 TI - Peripheral blood intracellular cytokine analysis in children newly diagnosed with inflammatory bowel disease. AB - Patterns of cytokine profiles have emerged for different forms of inflammatory bowel disease with a predominance of type 1 cytokines in patients with Crohn disease and type 2 cytokine expression in patients with ulcerative colitis. Most of these studies have involved older patients with long-standing disease or after various therapeutic interventions, and patterns of cytokine expression were hypothesized to be influenced by these factors. To evaluate for these possibilities, we studied 23 patients (15 boys) with newly diagnosed Crohn disease (n = 14) or ulcerative colitis. Their mean age at diagnosis was 13.1 +/- 2.9 y (mean +/- SD). Healthy control subjects (n = 9) were previously obtained. Peripheral blood intracellular cytokine analysis was performed within 24 h using a modification of Becton Dickinson's FastImmune Cytokine system. Multiparametric flow cytometry and phenotyping of lymphocytes was performed. T-cell populations were defined as type 1 being CD69(+), CD3(+), and interferon-gamma(+) and type 2 being CD69(+), CD3(+), and IL-4(+). The median percent of type 1 T cells from normal subjects (2.8%) was similar to that of ulcerative colitis subjects (1.8%, p > 0.20) but greater than that of Crohn disease subjects (0.55%, p = 0.05). The median percent of type 2 lymphocytes in normal subjects (1.8%) was greater than that of ulcerative colitis subjects (0.35%, p = 0.02) but was similar to that of Crohn disease subjects (1.1%, p > 0.20). Serial determinations showed the median percent of type 2 T cells increased in ulcerative colitis patients as remission was induced. Reduced activated peripheral type 1 T cells of newly diagnosed, untreated children are similar to interferon-gamma expression in mucosa of adults with postoperative recurrence. Reduced type 2 cytokine expression patterns in subjects with ulcerative colitis are similar to lamina propria T-cell expression levels in adults and improve with disease remission. PMID- 11861939 TI - Different acute-phase response in newborns and infants undergoing surgery. AB - In a prospective clinical study, we investigated the inflammatory response in 88 neonatal subjects (43 boys and 45 girls) who underwent major abdominal surgery owing to congenital malformation involving the gastrointestinal tract and compared it with the response in 20 infants (8 boys, 12 girls; mean age, 4 mo) who underwent elective surgery for resolution of an existing temporary stoma. In both groups, plasma levels of endotoxin, IL-6, and C-reactive protein as well as leukocyte counts were determined during and after surgery. Endotoxin was measured by the Limulus amebocyte test, IL-6 by ELISA, and C-reactive protein by nephelometry. Statistical analyses were performed using the Wilcoxon signed-rank test. A significant increase in circulating endotoxin and a leukocyte shift was observed in the infant group only. Postoperatively, IL-6 levels peaked between 2 and 6 h and C-reactive protein after 24 h in the infant group. In contrast, no significant increase in the levels of endotoxin, IL-6, and C-reactive protein in plasma were observed during and after surgery in the neonatal subjects, except those with gastroschisis. Newborns with gastroschisis developed an inflammatory response after surgery that was less pronounced than the response of infants older than 4 mo. The finding that endotoxemia in newborns does not follow surgical trauma is most likely because of the absence of bacterial colonization of the gastrointestinal tract. PMID- 11861940 TI - Direct measurement of intratracheal pressure in pediatric respiratory monitoring. AB - We describe a method based on a Fabry-Perot interferometer at the tip of an optic fiber with a diameter of 0.25 mm for direct measurement of tracheal pressure in pediatric respiratory monitoring. The response time of the pressure transducer and its influence on the resistance of pediatric endotracheal tubes (internal diameter, 2.5 to 5 mm) during constant and dynamic flow at different ventilator settings in a lung model were measured. The transducer was positioned at -1.5 (inside), 0, and +1.5 cm (outside) relative to the tip of the endotracheal tube and compared with a reference pressure inside the trachea. The clinical application of the transducer was tested in five pediatric patients. The response time of the transducer was 1.3 ms. The influence of the fiberoptic transducer on tube resistance was negligible during constant flow in inspiratory and expiratory directions for all endotracheal tubes tested. There was no difference in pressure measurements with the transducer positioned at or 1.5 cm below or above the tip of the endotracheal tube during dynamic measurements. During clinical circumstances insertion of the fiberoptic transducer was easy, recordings were stable, and the safety of the patient was not jeopardized. The fiberoptic transducer provided a reliable and promising way of monitoring tracheal pressure in intubated pediatric patients. The presence of the probe did not interfere with either pressure-flow relationship or patient care and safety. The technique is proposed for monitoring of respiratory mechanics and calculation of changes in tube resistance caused by kinking and secretions. PMID- 11861941 TI - CoASH and CoASSG levels in lungs of hyperoxic rats as potential biomarkers of intramitochondrial oxidant stresses. AB - Coenzyme A (CoASH) is compartmentalized preferentially in the mitochondria, and CoASH and its mixed disulfide with glutathione (CoASSG) undergo thiol/disulfide exchange reactions with glutathione (GSH) and glutathione disulfide (GSSG) in vitro. We measured CoASH and CoASSG in freeze-clamped lung tissues from Fischer 344 and Sprague-Dawley rats maintained in room air or exposed to >95% O(2) for 48 h to test the hypothesis that oxidant stresses on lung thiol status would be observed in the CoASH/CoASSG redox couple, suggesting oxidant stress responses in the mitochondria. Lung tissue concentrations of CoASSG in the Fischer-344 rats declined from 0.89 +/- 0.15 to 0.51 +/- 0.13 nmol/g of lung after 48 h of hyperoxia. CoASH levels declined from 6.40 +/- 0.84 to 3.0 +/- 0.65 nmol/g of lung, and acetyl CoA levels also were lower in the lungs of animals exposed to hyperoxia. CoASH/CoASSG ratios were lower in animals exposed to hyperoxia, satisfying our previously defined criteria for an oxidant stress on this thiol/disulfide redox couple, but absolute CoASSG levels were not increased, as would be expected for oxidant stresses driven simply by increases in reactive oxygen species or other oxidants. Pulmonary edema was observed in the hyperoxic rats and accounted for some of the declines in CoASH concentrations, but CoASH contents per total lung also declined. Lung mitochondrial succinate dehydrogenase activities were not diminished in rats exposed to hyperoxia, indicating that the decreases in CoASH concentrations are not attributable to general destruction of lung mitochondria. Lung GSSG contents were greater in the hyperoxia animals, but GSH/GSSG ratios, which are dominated by extramitochondrial pools, did not decrease in these animals. The mechanisms responsible for, and the possible pathophysiologic consequences of, the decreases in lung CoASH concentrations are not evident from the data available at the present time, but the loss of more than half the tissue contents of CoASH is likely to generate additional metabolic effects that could have significant pathophysiologic consequences. PMID- 11861942 TI - Delayed postischemic hypothermia improves long-term behavioral outcome after cerebral hypoxia-ischemia in neonatal rats. AB - Hypothermia may be an ideal neuroprotective intervention in hypoxic-ischemic encephalopathy after perinatal asphyxia. The present study describes the long term effects of prolonged resuscitative whole-body hypothermia initiated 2 h after hypoxic-ischemic injury on brain morphology and neuropsychological behavior in 7-d-old rats. After right common carotid artery ligation and exposure to hypoxia of 8% O(2) for 105 min, 10 animals were kept normothermic at 37 degrees C and 10 animals were cooled to 30 degrees C rectal temperature for 26 h, starting 2 h after the hypoxic-ischemic insult. All hypoxic-ischemic animals were gavage fed to guarantee long-term survival. Neuroprotection was evaluated by magnetic resonance imaging and behavioral testing. Hypothermia significantly reduced the final size of cerebral infarction by 23% at 6 wk after the insult. The most extended tissue rescue was found in the hippocampus (21%, p = 0.031), followed by the striatum (13%, p = 0.143) and the cortex (11%, p = 0.160). Cooling salvaged spatial memory deficits verified at 5 wk of recovery with Morris Water Maze test; whereas circling abnormalities after apomorphine injection and sensory motor dysfunctions on rotating treadmill improved, yet did not reach statistical significance. When compared with controls, hypoxic-ischemic animals performed worse in all behavioral tests. Hypothermia did not influence functional outcome in controls. Significant correlations between behavioral performance and corresponding regional brain volumes were found. We conclude that 26 h of mild to moderate resuscitative hypothermia leads not only to brain tissue rescue, but most important to long-lasting behavioral improvement throughout brain maturation despite severity of injury and delayed onset of cooling. PMID- 11861943 TI - Fate of (125)I-IGF-I administered into the amniotic fluid of late-gestation fetal sheep. AB - Large amounts of amniotic fluid (AF) are swallowed in late gestation. AF is the most accessible fetal compartment and provides a possible paraplacental route for the therapeutic administration of hormones and nutrients to the fetus. We therefore wished to investigate the fate of the predominant fetal growth factor, IGF-I, administered into AF of late-gestation ovine fetuses. Seven chronically catheterized fetuses at 124 d gestation had approximately 800 x 10(6) dpm of (125)I-IGF-I injected into the AF. AF and blood samples were withdrawn for up to 6 d. At 131 d gestation a postmortem examination was performed. All AF, blood, and tissue samples were counted. Selected samples of AF, blood, and gut contents underwent size-separation chromatography. (125)I-IGF-I was rapidly mixed in AF, with a significant difference in counts from different regions of the cavity persisting for only 3 h (p < 0.05). In vivo binding of (125)I-IGF-I in AF correlated highly with AF IGF binding protein 3 concentrations (r(2) = 0.93, p < 0.0001). In some animals, free (125)I-IGF-I persisted in AF and in plasma for the duration of the experiments. Chromatography of plasma samples demonstrated that intact (125)I-IGF-I was taken up from the fetal gut. Only fetal gut and thyroid contained appreciable counts at postmortem examination. Gut contents had more counts than gut wall, and the number of counts in gut contents increased distally (p < 0.05 for colon contents versus other regions). We conclude that there is sustained delivery of (125)I-IGF-I from the AF to the gut and systemic circulation of the ovine fetus after a single intraamniotic injection. PMID- 11861944 TI - Oral bovine serum concentrate improves cryptosporidial enteritis in calves. AB - Cryptosporidium parvum produces a prolonged watery diarrhea unresponsive to conventional antimicrobials. Because of reported efficacy of antibody-based immunotherapy, we studied the effect of inexpensive, commercially available oral bovine serum concentrate (BSC) in experimental cryptosporidiosis. Twenty-four calves were treated with 57 g/d BSC (n = 12) or soy protein (n = 12) added to their standard whey protein-based milk replacer (227 g/2 L twice daily). Of the 24, 9 were also treated with L-glutamine (GLN), 8 g/L (50 mM) in the milk (5 calves in the BSC group and 4 in the soy group). Animals were inoculated with 10(8) cryptosporidium oocysts per os on d 8 of life and received oral rehydration on d 12-14. Eight uninfected controls were treated with BSC or soy protein. Fecal and urine volume and urinary Cr-EDTA excretion were measured. Animals were killed on d 18 of life. Cryptosporidiosis induced severe watery diarrhea lasting >9 d and produced a 25% increase in intestinal permeability, a 33% decrease in villous surface area, and a 40% reduction in mucosal lactase specific activity. Glutamine treatment had no effect on the diarrhea or any of the intestinal tests; and therefore pooled data were used to compare the 12 calves treated with BSC with the 12 treated with soy. In animals receiving BSC, peak diarrheal volume and intestinal permeability were reduced 33%, fewer oocysts were shed, intestinal crypts were significantly deeper, and villous surface area returned to normal by 9 d after infection (all p > CPF >> CPF-oxon; in humans CPF-oxon was not quantifiable. Simulations were compared against experimental data and previously published studies in rats and humans. The model was utilized to quantitatively compare dosimetry and dynamic response between rats and humans over a range of CPF doses. The time course of CPF and TCP in both species was linear over the dose range evaluated, and the model reasonably simulated the dose-dependent inhibition of plasma ChE, RBC acetylcholinesterase (AChE), and brain (rat only) AChE. Model simulations suggest that rats exhibit greater metabolism of CPF to CPF-oxon than humans do, and that the depletion of nontarget B-esterase is associated with a nonlinear, dose-dependent increase in CPF-oxon blood and brain concentration. This CPF PBPK/PD model quantitatively estimates target tissue dosimetry and AChE inhibition and is a strong framework for further organophosphate (OP) model development and for refining a biologically based risk assessment for exposure to CPF under a variety of scenarios. PMID- 11861972 TI - Nonadditive hepatic tumor promoting effects by a mixture of two structurally different polychlorinated biphenyls in female rat livers. AB - This study evaluates and quantifies the interactive hepatic tumor promoting effects of two PCBs, the Ah receptor agonist PCB 126 (3,3',4,4',5 pentachlorobiphenyl) and the constitutive androstane receptor (CAR) agonist PCB 153 (2,2',4,4',5,5'-hexachlorobiphenyl). Promotion of altered hepatic foci was evaluated utilizing a medium-term 8-week bioassay for promoters of hepatocarcinogenesis. The assay employs placental glutathione-S-transferase positive (GST-P+) liver cell foci as markers of preneoplasia in female Fischer 344 rats treated with the known initiator diethylnitrosamine followed by partial hepatectomy and by gavage exposure to test chemicals. GST-P+ foci were quantified by histomorphometry and were reported as areas and numbers of GST-P+ foci within the area of liver examined. For PCB 126, the doses were 0.1, 1.0, and 10 microg/kg body weight. For PCB 153, the doses were 10, 100, 1000, 5000, and 10,000 microg/kg body weight. Combined PCB 126 and 153 exposures were 0.1 + 10, 1 + 100, 10 + 1000, 10 + 5000, and 10 + 10,000 microg/kg, respectively. Individual PCB treatment resulted in dose dependent increases in liver and adipose concentrations. Hepatic PCB 153 levels were significantly increased (p < 0.01) after combined exposure. Treatment with PCB 126 or PCB 153 alone resulted in a significant (p < 0.01) dose dependent increase in GST-P+ foci area and number compared with controls. Treatment with the mixture of PCB 126 and 153 resulted in antagonistic GST-P+ focus formation (p < 0.001) for both foci area and number. The less than additive effect was present at all 5 PCB 126/PCB 153 dose combinations, including the low doses of PCB 126 and 153 that did not show significant promotional activity alone. PMID- 11861973 TI - Dioxin inhibition of estrogen-induced mouse uterine epithelial mitogenesis involves changes in cyclin and transforming growth factor-beta expression. AB - A single dose of dioxin (2,3,7,8-tetrachlorodibenzo-p-dioxin or TCDD; 5 microg/kg, ip) inhibits 17beta-estradiol (E2)-induced uterine epithelial mitogenesis, apparently through disruption of stromal-epithelial interactions. To understand if TCDD alters early uterine (Ut) responses to E2, young adult C57BL/6J mice were ovariectomized and given (i.p.) either oil or 5 microg/kg TCDD. After 24 h, TCDD-treated mice received E2, and oil-treated mice were given E2 or oil. Body and Ut weights were collected 6 and 18 h later. Ut were flash frozen at 6 h. E2 increased Ut weight (p < 0.0001) and Ut/body weight ratio (p < 0.0001), compared to mice given oil alone. Ut cyclin expression was assessed by an RNase protection assay. E2 increased mRNA expression for cyclin A2 and B1 (p < 0.05), in addition to D1, D2, and D3 (p < 0.001), while cyclin C was unchanged from oil controls and cyclins A1 and B2 were undetectable. In contrast, TCDD completely abolished E2-induced cyclin A2, which has been associated with S phase initiation, and reduced B1 and D2 (p < 0.05). Interestingly, TCDD did not alter E2-induced Ut weight increases at 6 h, but inhibited E2-induced Ut weight gain at 18 h. A 10-microg/kg TCDD dose was necessary for attenuation of the early E2 induced Ut weight increases (p < 0.01). Since TGF-beta regulates cyclins, Ut TGF beta was also assessed in TCDD + E2-treated and control mice. TGF-beta mRNA levels were increased after TCDD compared to E2 alone (p < 0.01), suggesting a possible mechanism for TCDD inhibition of Ut cyclin A2. Thus, TCDD alters specific E2-regulated Ut G(1) phase activities and may inhibit E2-induced Ut epithelial mitogenesis by disrupting specific cell signaling mechanisms necessary for S phase initiation in vivo. PMID- 11861974 TI - A novel cell line, MDA-kb2, that stably expresses an androgen- and glucocorticoid responsive reporter for the detection of hormone receptor agonists and antagonists. AB - The U.S. Environmental Protection Agency has proposed that in vitro assays for estrogen receptor (ER)- and androgen receptor (AR)-mediated actions be included in a Tier-I screening battery to detect hormonally active chemicals. Herein we describe the development of a novel stable cell line, MDA-kb2, for screening of androgen agonist and antagonists and to characterize its specificity and sensitivity to endocrine-disrupting chemicals. The breast cancer cell line, MDA MB-453, was stably transformed with the MMTV.luciferase.neo reporter gene construct. Since both GR and AR are present in the MDA-MB-453 cells, and both receptors can act through the MMTV promoter, compounds that act through either AR or GR activate the MMTV luciferase reporter. As expected, AR agonists such as dihydrotestosterone (DHT), and GR agonists such as dexamethasone (DEX), corticosterone, and aldosterone induce luciferase expression at appropriate concentrations. DHT consistently produced 3-9-fold induction at concentrations from 0.1 to 10 nM. At 1 to 1000 nM, DEX induced luciferase activity 1.3-19.5 fold. To distinguish AR- from GR-mediated ligands, chemicals were assayed concurrently with the antiandrogen, hydroxyflutamide (OHF), which blocks AR- but not GR-mediated responses. In addition, known AR antagonists, including hydroxyflutamide, vinclozolin, vinclozolin metabolites M1 and M2, p,p'-DDE, and linuron inhibited DHT-induced luciferase gene expression at appropriate concentrations in this system. We have found that these cells are relatively easy to culture and maintain. Responsiveness was monitored over time and was stable for more than 80 passages. Some advantages of this assay are that it is relatively rapid (2 days), eliminates the need for transfection, can be conducted in a 96-well plate format, and produces consistent reproducible results. In summary, we have developed a cell line that can be used to screen chemicals, not just for AR- but for GR-mediated activities as well. PMID- 11861975 TI - Development of two androgen receptor assays using adenoviral transduction of MMTV luc reporter and/or hAR for endocrine screening. AB - The discovery of xenobiotics that interfere with androgen activity has highlighted the need to assess chemicals for their ability to modulate dihydrotestosterone (DHT)-receptor binding. Previous test systems have used cells transfected with plasmid containing a reporter gene. Here we report the use of transduction for gene delivery and assessment of the modulation of DHT-induced gene activation. Transduction, the ability of replication-defective viruses to deliver biologically competent genes, is a well understood biological process, which has been utilized to repair defective genes in humans as well as to express exogenous genes in rodent models. Human breast carcinoma cells (MDA-MB-453) containing endogenous copies of the androgen (hAR) and glucocorticoid (GR) receptors were transduced with replication-defective human adenovirus type 5 containing the luciferase (Luc) reporter gene driven by the AR- and GR-responsive glucocorticoid-inducible hormone response element found with the mammary tumor virus LTR (Ad/MLUC7). In a second set of experiments, CV-1 cells were transduced as above with MMTV-luc and also hAR. Cells were subcultured in 96-well plates, transduced with virus, exposed to chemicals, incubated for 48 h, lysed, and assayed for luciferase. Luc gene expression was induced in a dose-dependent manner by DHT, estradiol, and dexamethasone (MDA only) and inhibited by AR antagonist hydroxyflutamide (OHF), hydroxy-DDE, HPTE (2,2-bis(p-hydroxyphenyl) 1,1, 1-trichloroethane), a methoxychlor metabolite, and M1 and M2 (vinclozolin metabolites). The transduced cells responded to AR agonists and antagonists as predicted from our other studies, with a very robust and reproducible response. Over all replicates, 0.1 nM DHT induced luc expression by about 45-fold in CV-1 cells (intra-assay CV = 20%) and 1micromolar OHF inhibited DHT by about 80%. In the transduced MDA cells, 0.1 nM DHT induced luc by about 24-fold (intra-assay CV = 33%), which was inhibited by OHF by about 85%. DHT-induced luciferase activity peaked in both cell lines between 1 and 100 nM, displaying about 64- and 115-fold maximal induction in the CV-1 and MDA 453 cells, respectively. For agonists, a two-fold induction of luc over media control was statistically significant. For AR antagonists, a 25-30% inhibition of DHT-induced luc expression was typically statistically significant. Comparing the two assays, the transduced CV-1 cells were slightly more sensitive to AR-mediated responses, but the transduced MDA 453 cells were more responsive to GR agonists. In summary, these assays correctly identified the endocrine activity of all chemicals examined and displayed sensitivity with a relatively low variability and a high-fold induction over background. Adenovirus transduction for EDC screening has the potential to be employed in a high-throughput mode, and could easily be applied to other cell lines and utilized to deliver other receptors and reporter genes. PMID- 11861976 TI - Combined effects of dietary phytoestrogen and synthetic endocrine-active compound on reproductive development in Sprague-Dawley rats: genistein and methoxychlor. AB - Humans and wildlife are frequently exposed to mixtures of endocrine active compounds (EAC). The objective of the present study was to investigate the potential of the phytoestrogen genistein to influence the reproductive developmental toxicity of the endocrine-active pesticide methoxychlor. Three levels of genistein (0, 300, or 800 ppm) and two levels of methoxychlor (0 or 800 ppm) were used in this study. Sprague-Dawley rats were exposed to the two compounds, either alone or in combinations, through dietary administration to dams during pregnancy and lactation and to the offspring directly after weaning. Both compounds, methoxychlor in particular, were associated with reduced body growth at 800 ppm, but pregnancy outcome was not affected by either treatment. An acceleration of vaginal opening (VO) in the exposed female offspring was the only observed effect of genistein at 300 ppm. Exposure to 800 ppm genistein or 800 ppm methoxychlor caused accelerated VO and also altered estrous cyclicity toward persistent estrus in the female offspring. The estrogenic responses to genistein and methoxychlor administered together were apparently accumulative of the effects associated with each compound alone. Methoxychlor, but not genistein, delayed preputial separation (PPS) in the male rats. When administered with methoxychlor, genistein at 800 ppm enhanced the effect of methoxychlor on delaying PPS. Genistein and methoxychlor treatment did not change gender-specific motor activity patterns in either sex. To explore possible mechanisms for interaction between the two compounds on development, we performed estrogen receptor (ER)- and androgen receptor (AR)-based in vitro transcriptional activation assays using genistein and the primary methoxychlor metabolite 2,2-bis (p-hydroxyphenyl)-1,1,1-trichloroethane (HPTE). While the in vitro assays supported the estrogenic effects of genistein and methoxychlor and the antiandrogenic effects of methoxychlor, the reactivity of these compounds with ERs alpha and beta could not predict the greater in vivo estrogenic potency of methoxychlor over genistein; nor could the potentiation of the methoxychlor effect on PPS by genistein be predicted based on in vitro HPTE and genistein reactions with the AR. Data from this study indicate that phytoestrogens are capable of altering the toxicological behaviors of other EACs, and the interactions of these compounds may involve complexities that are difficult to predict based on their in vitro steroid receptor reactivities. PMID- 11861977 TI - Developmental exposure to brominated diphenyl ethers results in thyroid hormone disruption. AB - The objective of the current study was to characterize the effects of DE-71 (a commercial polybrominated diphenyl ether mixture containing mostly tetra- and penta-bromodiphenyl ethers) on thyroid hormones and hepatic enzyme activity in offspring, following perinatal maternal exposure. Primiparous Long-Evans rats were orally administered DE-71 (0, 1, 10, and 30 mg/kg/day) in corn oil from gestation day (GD) 6 to postnatal day (PND) 21. Serum and liver samples obtained from dams (GD 20 and PND 22), fetuses (GD 20), and offspring (PNDs 4, 14, 36, and 90) were analyzed for circulating total serum thyroxine (T(4)) and triiodothyronine (T(3)), or hepatic microsomal ethoxy- and pentoxy-resorufin-O deethylase (EROD and PROD), and uridine diphosphoglucuronosyl transferase (UDPGT) activity. There were no significant effects of treatment on maternal body weight gain, litter size, or sex ratio, nor were there any effects on any measures of offspring viability or growth. Serum T(4) was reduced in a dose-dependent manner in fetuses on GD 20 (at least 15%) and offspring on PND 4 and PND 14 (50 and 64% maximal in the 10 and 30 mg/kg/day groups, respectively), but recovered to control levels by PND 36. Reduction in serum T(4) was also noted in GD 20 dams (48% at highest dose), as well as PND 22 dams (44% at highest dose). There was no significant effect of DE 71 on T(3) concentrations at any time in the dams or the offspring. Increased liver to body weight ratios in offspring were consistent with induction of EROD (maximal 95-fold), PROD (maximal 26-fold) or UDPGT (maximal 4.7-fold). Induction of PROD was similar in both dams and offspring; however, EROD and UDPGT induction were much greater in offspring compared to dams (EROD = 3.8-fold; UDPGT = 0.5-fold). These data support the conclusion that DE-71 is an endocrine disrupter in rats during development. PMID- 11861978 TI - Effect of a single oral dose of 2,3,7,8-tetrachlorodibenzo-p-dioxin on immune function in male NC/Nga mice. AB - Exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) induces immunosuppression in humans and animals. However, the effect of TCDD on Th2-type immune responses such as allergic reactions has been unclear. Using NC/Nga mice that developed atopic dermatitis-like skin lesions with marked elevation in plasma of total IgE when bred under conventional conditions, we investigated the effects of a single oral dose of TCDD on immune responses. NC/Nga mice received a single oral dose (0 or 20 microg/kg body weight) of TCDD. On day 7, treatment with TCDD alone decreased the cellularity of thymus. However, treatment with TCDD modified the cellularity of spleens and mesenteric lymph nodes (MLNs) but not of the thymus on day 28. When NC/Nga mice received ip immunization with OVA and alum on the same day as the TCDD treatment (0, 5, or 20 microg/kg body weight), TCDD markedly suppressed the concentrations of Th2-type cytokines (e.g., IL-4 and IL-5) in culture supernatants of spleen cells, whereas IFN-gamma production significantly increased. TCDD exposure reduced anti-OVA and total IgE antibody titers in plasma and did not induce the development of atopic dermatitis-like lesions in the pinnae or dorsal skin of NC/Nga mice. These results suggest that in NC/Nga mice, exposure to TCDD may impair the induction of Th2-type immune responses. PMID- 11861979 TI - Effect of varying caloric restriction levels on female rat growth and 5 hydroxymethyl-2'-deoxyuridine in DNA. AB - Caloric restriction has previously been shown to decrease levels of oxidative stress in rats. In this study, we examined the effects of 5 different caloric intake levels on one type of oxidative DNA damage in rat mammary gland, blood, and liver. Animals were fed modified AIN-93G diets to accommodate 10, 20, 30, or 40% calorie restriction (CR), relative to ad libitum (AL) consumption. The intakes of fat, protein, vitamins, and minerals thus remained constant, but total carbohydrate intake decreased. Body weights of the animals at 20 weeks reflected the degree of restriction, but in the first 10 weeks, weight gain in the 10% CR group was not reduced relative to animals fed ad libitum. Levels of 5 hydroxymethyl-2'-deoxyuridine increased with time in mammary gland and nucleated blood cells regardless of CR level, indicating an effect of animal age, despite the fact that the animals were only 7 months old after the 20-week dietary study. In liver, however, there was a trend towards decreased DNA damage levels with time. The effect of diet on levels of 5-hydroxymethyl-2'-deoxyuridine was not statistically significant, indicating no protective effect of restricted dietary carbohydrate. This dietary study differed from previous work in that the modified AIN-93G dietary formulation contains relatively higher levels of fat and vitamins K, E, and B(12), and it has certain added trace minerals. This data raises the question of whether the previously reported effects of calorie restriction on preventing oxidative stress in mammary gland are dependent on the type of dietary formulation used. PMID- 11861980 TI - Potentiation of noise-induced hearing loss by low concentrations of hydrogen cyanide in rats. AB - Noise-induced hearing loss is the most prevalent occupational injury in the United States despite the adoption of clear permissible exposure limits and protocols for hearing conservation. This study identifies low-level chemical asphyxiant exposure as a risk factor capable of potentiating noise-induced hearing loss. Rats were exposed to 10, 30, and 50 ppm hydrogen cyanide (HCN) alone for 3.5 h (n = 28) or in combination with 2 h octave band noise exposure (100 dB(lin); n = 28). Additional groups received noise exposure alone (n = 16) and no treatment other than placement in an inhalation chamber with clean air and quiet (n = 16). Pure tone compound action potential (CAP) thresholds were determined 4 weeks following the exposure in order to assess pure tone auditory sensitivity and permanent threshold impairment. Cochleae from an additional 13 subjects were processed for light microscopy to permit assessment of hair cell loss. The results demonstrate that the noise exposure alone impaired CAP threshold by about 10 dB, averaged between 12-40 kHz, and produced a 5% loss of outer hair cells at the base of the cochlea, but no inner hair cell loss. The combined exposure to noise and HCN caused a cyanide dose-dependent CAP threshold impairment that exceeds the noise exposure alone. This effect reached statistical significance at a HCN level of 30 ppm. Combined exposure also produced more outer hair cell loss than noise alone. HCN alone did not cause significant hearing loss or hair cell loss. A risk assessment analysis was conducted for the auditory threshold data using benchmark dose software published by the U. S. EPA (BMDS version 1.3). A continuous model showed that the data could be described by a linear function. For a benchmark response corresponding to a 5 dB increase in auditory threshold above the effect of noise alone, the lower bound on the 95% confidence interval for the benchmark dose was 9 ppm. The benchmark dose that impaired auditory threshold 10% above the effect of noise alone had a lower bound of 2 ppm. The lower bound to the HCN dose that produced a 1 SD elevation in noise induced hearing loss was 16 ppm. These exposure levels provide a range of concentrations below to slightly above the short-term exposure limit for HCN. However, if these levels are adjusted for an 8 h time-weighted average (TWA), the resulting levels are below the permissible exposure level (PEL) for HCN. PMID- 11861981 TI - Rat hippocampal glutamate and GABA release exhibit biphasic effects as a function of chronic lead exposure level. AB - Previous work has suggested that the lead (Pb) exposure-induced decrease in K(+) evoked hippocampal glutamate (GLU) release is an important factor in the elevated threshold and diminished magnitude reported for hippocampal long-term potentiation (LTP) in exposed animals. In addition, complex dose-effect relationships between Pb exposure level and LTP have been reported. This investigation was conducted to determine the effects of Pb on hippocampal GLU and GABA release as a function of exposure level. Rats were continuously exposed to 0.1, 0.2, 0.5, or 1.0% Pb in the drinking water beginning at gestational day 15 16. Hippocampal transmitter release was induced in adult males by perfusion of 150 mM K(+) in the presence of Ca(+2) (total release) through a microdialysis probe in one test session, followed by perfusion through a contralateral probe in the absence of Ca(+2) (Ca(+2)-independent release) in the second session. Chronic exposure produced decreases in total K(+)-stimulated hippocampal GLU and GABA release at exposure levels of 0.1-0.5% Pb. Maximal effects were seen in the 0.2% group (blood Pb = 40 microg/100 ml), and changes in total release could be directly traced to alterations in the Ca(+2)-dependent component. However, these effects were less evident in the 0.5% group and were no longer present in the 1.0% Pb group, thus defining U-shaped dose-effect relationships. Moreover, in the absence of Ca(+2) in the dialysis perfusate, K(+)-induced release was elevated in the 2 highest exposure groups, suggesting a Pb(+2)-induced enhancement in evoked release. This pattern of results indicates the presence of 2 actions of Pb on in vivo transmitter release: a more potent suppression of stimulated release seen at lower exposure levels (27-62 microg/100 ml) combined with Ca+2-mimetic actions to independently induce exocytosis that is exhibited at higher exposure levels (> or =62 microg/100 ml). Furthermore, significant similarities in the dose-effect relationships uncovered in measures of evoked GLU release and hippocampal LTP (M. E. Gilbert et al., 1999b, Neurotoxicology 20, 71-82) reinforce the conclusion that exposure-related changes in GLU release play a significant role in the Pb induced effects seen in this model of synaptic plasticity. PMID- 11861982 TI - Sensorimotor deficit and cholinergic changes following coexposure with pyridostigmine bromide and sarin in rats. AB - A myriad of neurological symptoms including muscle and joint pain, ataxia, chronic fatigue, headache, and difficulty in concentration have been reported by Persian Gulf War (PGW) veterans. A large number of these veterans were prophylactically treated with pyridostigmine bromide (PB) and possibly exposed to sarin. In the present study we investigated the effects of PB and sarin, alone and in combination, on sensorimotor performance and the central cholinergic system of rats. Male Sprague-Dawley rats were treated with PB (1.3 mg/kg, 15 daily doses, oral) and sarin (50, 75, 90, and 100 microg/kg, single im dose on day 15), alone and in combination. The animals were evaluated for postural reflexes, limb placing, orienting to vibrissae touch, incline plane performance, beam-walk time, and forepaw grip time 7 and 15 days following treatment with sarin. Treatment with either PB or sarin alone resulted in significant sensorimotor impairments. Coexposure to sarin and PB resulted in significant sensorimotor deficits that worsened over time. By 15 days following sarin treatment, plasma butyrylcholinesterase (BChE) activity returned to normal levels in the animals treated with sarin alone, whereas in the animals exposed to PB or PB plus sarin, there was an increase in the enzyme activity. Cortical acetylcholinesterase (AChE) activity remained inhibited in the animals treated with sarin alone and in combination with PB. Muscarinic acetylcholine receptor (m2 mAChR) ligand binding with [(3)H]AFDX-384 in cortex and brain stem showed significant increases (approximately 120-130% of control) following coexposure to PB and sarin at higher doses. To evaluate the potential of PB for augmentation or inhibition of the toxicity induced by acute sarin exposure, the animals were exposed to either 10 or 100 microg/kg sarin (single im injection) with or without pretreatment with PB, and sacrificed 3 h after treatment with sarin. Pretreatment with PB offered slight protection in the plasma as well as brain regional enzyme activities. Pretreatment with PB did not have any effect on sarin-inhibited brain regional AChE activity following treatment with 100 microg/kg sarin. These results show that prophylactic treatment with PB offers some degree of protection in peripheral cholinesterase. Furthermore, these results show that treatment with either sarin or PB alone resulted in sensorimotor impairments, while coexposure to high doses of sarin with PB caused an exacerbated deficit. PMID- 11861983 TI - Selective inhibition of murine palatal mesenchymal cell proliferation in vitro by secalonic acid D. AB - Secalonic acid D (SAD), a teratogenic mycotoxin, induces cleft palate (CP) in the offspring of exposed mice by inhibiting palatal shelf growth. Since reduced proliferation, increased apoptosis, and/or decreased extracellular matrix (ECM) synthesis of palatal mesenchymal cells (PMC) can all contribute to smaller shelf size, the hypothesis that teratogenically relevant concentrations (0 to 120 microg/ml) of SAD will have adverse effects on one or more of these cellular processes was tested, using primary murine PMC cultures. Exposure to SAD resulted in significant and dose-dependent decreases in mesenchymal cell number, uptake of (3)H-thymidine, and expression of proliferating cell nuclear antigen (PCNA). Trypan blue dye exclusion assay, however, revealed significant cell death only at higher doses, suggesting that the decrease in cell number at lower (more realistic) doses is likely a consequence of reduced cell proliferation and not cell death. Further, negative results in the DNA fragmentation analysis following SAD exposure suggested that cell death caused by higher levels of SAD was unrelated to apoptosis. Similarly, results of (3)H-glucosamine uptake assay indicated inhibitory effect of SAD on accumulation of hyaluronic acid (HA) or sulfated glycosaminoglycans (sGAG) only at the highest dose tested. Also, SAD affected neither extracellular nor cell-associated fibronectin expression at any dose tested. Taken together, these data suggest that the pathogenesis of CP by SAD is likely a result of a reduction in the size of the palatal shelf caused by SAD-induced inhibition of mesenchymal cell proliferation. PMID- 11861984 TI - Muscarinic receptor antagonist-induced lenticular opacity in rats. AB - Investigations on compound A, an M2-sparing M3 muscarinic receptor antagonist, showed that focal polar anterior subcapsular lenticular opacities, characterized by focal epithelial proliferation, developed in Sprague-Dawley rats. The incidence and bilateral localization of this change increased generally with dose and time, though plateauing after 8 months of treatment; however the severity progressed very slightly. Over a 1-year period, no anterior cortical lens fiber changes or other histological ocular changes developed. A decreased severity of the change and apoptosis suggested some regression after a 26-week recovery period. Two nonselective muscarinic receptor antagonists, atropine and tolterodine, induced similar lenticular changes in rats. A hypothesis in relation to an indirect effect of the drug, such as increased illumination of the lens due to mydriasis observed with all these compounds, was investigated and disproven. Because these opacities are induced by structurally unrelated muscarinic receptor antagonists (atropine and tolterodine), it is likely that these lenticular changes are the result of muscarinic receptor inhibition. However, hypotheses regarding a direct effect of the drug on muscarinic receptors in the lens epithelium, possibly mediated by drug and/or metabolite(s) in the aqueous humor and/or lens epithelium, remain to be investigated. This lenticular opacity is similar to that observed spontaneously in Sprague-Dawley rats, although the latter occur at a lower incidence. No such lenticular opacities have been reported in other animal species, including man, after treatment with muscarinic receptor antagonists. PMID- 11861986 TI - Diagnosing deep vein thrombosis in the lower extremity: correlation of clinical and duplex scan findings. AB - OBJECTIVE: To identify factors that predict a positive duplex scan examination result in patients with suspected deep vein thrombosis of the lower extremity. DESIGN: Retrospective study. SETTING: Vascular laboratory in a university teaching hospital. PATIENTS AND METHODS: The results of 345 lower extremity duplex venous scans performed between August 1994 and November 1998 were reviewed. All patients were in-patients referred from different specialties due to clinical suspicion of lower extremity deep vein thrombosis. Positive duplex scans were correlated with patients' demographic data (sex, age), medical history (history of malignancy, deep vein thrombosis, and pulmonary embolism) and clinical features (leg swelling, venous insufficiency, calf pain, and leg ulcer). Univariate analysis was performed using the Chi squared test. RESULTS: A total of 345 scans were performed for 313 patients. The mean age was 55 years (range, 19 92 years). Sixty-three patients (49 male, 14 female) had a positive scan, giving a yield of 18.3%. Four factors had a significant association with a positive scan: male sex (P=0.0102), history of malignancy (P=0.0040), history of deep vein thrombosis (P=0.0001), and history of pulmonary embolism (P=0.0265). CONCLUSIONS: Common presenting clinical features do not predict the result of ultrasonographic investigation for deep vein thrombosis. The chance of having a positive scan is significantly higher in male patients and those with a history of malignancy, deep vein thrombosis, or pulmonary embolism. PMID- 11861987 TI - Paediatric hepatoblastoma and hepatocellular carcinoma: retrospective study. AB - OBJECTIVES: To compare and contrast clinical characteristics and outcomes of hepatoblastoma or hepatocellular carcinoma in paediatric patients. DESIGN: Retrospective study. SETTING: University teaching hospital, Hong Kong. PATIENTS AND METHODS: Medical records of 22 paediatric patients with hepatoblastoma (n=11) or hepatocellular carcinoma (n=11) admitted to Queen Mary Hospital between 1989 and 2000 were reviewed. Data gathered included demographic data, results of liver function tests, hepatitis A, B, and C titres, and alpha-foetoprotein levels, and imaging studies including chest X-ray, ultrasound study, computed tomography scan, and magnetic resonance imaging/hepatic angiogram for tumour staging and resectability. RESULTS: The mean age of patients with hepatoblastoma was 18 months (range, 5 months to 3 years), while that of patients with hepatocellular carcinoma was 10.2 years (range, 2 to 16 years). Females predominated in the hepatoblastoma group (female:male, 8:3) and males in the hepatocellular carcinoma group (male:female, 10:1). None of the patients with hepatoblastoma were hepatitis B surface antigen positive, in contrast to 64% of the hepatocellular carcinoma group. Only 45% of the hepatocellular carcinomas were resectable at presentation and this figure remained unchanged following chemotherapy. A total of 91% of hepatoblastomas were resectable, four at presentation, and a further six after chemotherapy. Tumour rupture was more common in patients with hepatoblastoma than in those with hepatocellular carcinoma (36% versus 9% of cases, respectively). Mortality rates were considerably higher among the hepatocellular carcinoma group than the hepatoblastoma group in this series. CONCLUSION: Childhood hepatoblastoma and hepatocellular carcinoma differ with respect to age and tumour stage at presentation, hepatatis B surface antigen status, tendency to rupture, chemosensitivity, and prognosis. PMID- 11861988 TI - Subcutaneous steroid injection as treatment for chalazion: prospective case series. AB - OBJECTIVE: To study the efficacy of subcutaneous steroid injection in the treatment of chalazion. DESIGN: Prospective consecutive case series. SETTING: University teaching hospital, Hong Kong. PATIENTS: Patients with chalazion presenting to the out-patient clinic of the Department of Ophthalmology at the Prince of Wales Hospital from January to June 1998. MAIN OUTCOME MEASURES: Size of the chalazion after steroid injection treatment. RESULTS: Forty-eight consecutive patients with chalazion were treated with injection of triamcinolone into the subcutaneous tissue around the lesion. In 43 (89.6%) patients, the lesion subsided completely. Twenty-six (54.2%) patients had lesions that subsided with one injection. The size and duration of the chalazion at presentation did not significantly affect the outcome of the treatment. Two patients developed depigmentation of the skin at the site of injection. No other major complications were encountered. CONCLUSION: Subcutaneous injection of the steroid triamcinolone acetonide appears to be a simple and effective treatment for chalazion. Further comparative clinical trials are indicated. PMID- 11861989 TI - Picture archiving and communication system: prospective study. AB - OBJECTIVES: To evaluate the use of a picture archiving and communication system and user satisfaction in order to further improve its quality. DESIGN: Prospective study. SETTING: Medical college hospital, Japan. MATERIALS AND METHODS: An automated computerised method was used to collect the data from March 1999 to February 2000. MAIN OUTCOME MEASURES: Each workstation automatically recorded data on the rank of the user, purpose of use, use of postprocessing tools, and user satisfaction. RESULTS: The number of resident users in the radiology reading room increased and those outside the reading room decreased, but the number of staff users changed little. The purpose of use and the use of postprocessing functions in the reading room were not significantly different from those outside it (P=0.179 and P=0.269, respectively). The average numbers of images accessed to per workstation monthly in the reading room, the general practice ward, and the gastroenterology ward were 1081, 970 and 741, respectively. Only 12 images in the orthopaedic surgery out-patient clinic and 70 images in the orthopaedic surgery ward, however, were accessed per month. The percentages of satisfied users decreased both inside and outside the reading room. The degree of satisfaction of users in the reading room was significantly different from that outside it (P=0.004). The most common reason for dissatisfaction was the length of time necessary to retrieve images. CONCLUSIONS: It is necessary to shorten response times of picture archiving and communication system workstations. Repeated data collection and evaluation, however, are also important. PMID- 11861990 TI - Accelerated atherosclerosis in patients with systemic lupus erythematosus: a review of the causes and possible prevention. AB - Systemic lupus erythematosus is an autoimmune disorder affecting multiple organ systems. Patients with systemic lupus erythematosus exhibit a bimodal pattern of mortality, with those who have had the disease for 5 to 10 years being at increased risk of cardiovascular disease, particularly myocardial infarction. Elevated levels of conventional cardiovascular risk factors promote vascular damage resulting in impairment of normal endothelial function. In addition, autoantibodies directed against oxidised lipoproteins, along with chronic secretion of inflammatory cytokines and suppression of fibrinolytic parameters, are thought to increase atherogenesis. Treatment with corticosteroids may also contribute to the accelerated atherosclerosis observed in these patients. This review discusses the accentuated relationship between conventional cardiovascular risk factors, systemic lupus erythematosus-induced inflammatory changes and the early stages of atherogenesis and how careful monitoring of risk factors and use of appropriate therapies may reduce the progression of atheroma development in patients with systemic lupus erythematosus. PMID- 11861991 TI - Vertebrobasilar artery dissections: current practice. AB - Nine patients with vertebrobasilar artery dissections who presented with neurological symptoms or subarachnoid haemorrhage were identified and treated in the Neurosurgical Unit at the Prince of Wales Hospital in Hong Kong. An account of these patients is given and their treatment described. This paper further reviews selected literature to outline the concepts and appropriate management of vertebrobasilar artery dissections. PMID- 11861992 TI - Childhood fever revisited. AB - The academic study of body temperature began in 1868. Since then, a variety of thermometers have been developed for clinical use. A working knowledge of these different thermometers, the various sites for taking temperature, and the normal range of body temperatures, is essential. As the pathogenesis of fever is being elucidated, and the cytokines involved identified, the risks and benefits of fever to the organism are becoming better understood. The importance of recognising diagnostic fever patterns should be stressed. Acetaminophen (paracetamol) and ibuprofen are accepted as the standard medical treatments for fever. PMID- 11861993 TI - Topical chloramphenicol for eye infections. AB - Topical chloramphenicol has been widely used in the treatment and prevention of superficial eye infections due to its broad spectrum of activity and low cost. The use of this drug has decreased considerably in the United States since the first case of aplastic anaemia associated with topical chloramphenicol was reported in the 1960s. This medication, however, is still widely used in many other countries. This paper evaluates the evidence for and against the use of topical chloramphenicol in ocular diseases. PMID- 11861994 TI - Hepatocellular carcinoma with metastasis to the skull base, pituitary gland, sphenoid sinus, and cavernous sinus. AB - Two cases of hepatocellular carcinoma, with metastases to the skull base, pituitary gland, sphenoid sinus, and cavernous sinus are reported. Patients presented with diplopia, retro-orbital headache, and multiple cranial nerves palsies. Pituitary metastases may require surgery as palliative treatment, and for the confirmation of histology. One of the current cases was diagnosed with hepatocellular carcinoma prior to transphenoidal resection of the pituitary metastasis. The second patient was found to have hepatocellular carcinoma after review of histology, and the development of signs and symptoms relating to the primary tumour. PMID- 11861995 TI - Brain tuberculoma in Hong Kong. AB - We report two cases of brain tuberculoma occurring in patients residing in Hong Kong. Both patients presented with headache and had space-occupying lesions evident on computed tomography scans of the brain. The patients had no history of tuberculosis and no symptoms of concurrent extracranial tuberculosis were evident. The diagnosis of tuberculoma was made at the time of surgical excision. Delayed diagnosis of brain tuberculoma is likely to occur in industrialised countries where tuberculosis is rare. In Hong Kong, however, with a constant influx of foreign domestic workers from endemic regions, a high index of suspicion should be maintained. Imaging studies support, but do not confirm, the diagnosis of brain tuberculoma. We recommend obtaining a definitive histological diagnosis, with computed tomography-guided stereotactic biopsy, before starting antituberculous therapy. Surgical excision is necessary in patients with raised intracranial pressure secondary to the lesion, who are not responding to medical therapy. PMID- 11861996 TI - Prolonged jaundice following percutaneous coronary intervention and ticlopidine therapy. AB - Ticlopidine, an adenosine diphosphate receptor blocker, is widely used to prevent subacute stent thrombosis after percutaneous coronary intervention. Along with neutropenia and thrombotic thrombocytopenic purpura, cholestatic hepatitis is one of the most serious potential side-effects of ticlopidine therapy. Four patients with prolonged jaundice after ticlopidine therapy, including one fatal case, are presented. Alternative antithrombotic therapy for subsequent percutaneous coronary intervention is also described. Clopidogrel therapy was found to be safe and effective in two patients with a history of ticlopidine-related cholestatic hepatitis. PMID- 11861997 TI - Three patients with lead poisoning following use of a Chinese herbal pill. AB - We report on three patients with lead poisoning following use of the Chinese herbal pill Bao ning dan, prepared by the same traditional Chinese medicine practitioner. The patients had varying degrees of exposure to Bao ning dan and different clinical manifestations. Blood lead concentrations did not correlate with clinical severity. Two patients received chelating therapy and blood lead concentrations subsequently rapidly decreased. One patient was managed conservatively and end-organ complications resolved gradually. With increasing use of traditional Chinese medicines, related adverse reactions are expected to become increasingly common. Practitioners of western medicine should remain alert to this possibility. A comprehensive drug review, including the use of herbal medicines, should form a routine part of medical history taking. PMID- 11861998 TI - "You never told me that could happen!" Consent and litigation: how to avoid it. PMID- 11861999 TI - Improve patient safety and reduce medical errors. PMID- 11862000 TI - Universal screening of human immunodeficiency virus infection in pregnant women in Hong Kong. PMID- 11862002 TI - Regulation of innate immune responses by Toll-like receptors. AB - Innate immune response in Drosophila is mediated by signaling through Toll receptors. In mammals, Toll-like receptors (TLRs), comprising a large family, recognize a specific pattern of microbial components. So far, the roles of TLR2, TLR4, TLR5, TLR6, and TLR9 have been revealed. The recognition of microbial components by TLRs leads to activation of innate immunity, which provokes inflammatory responses and finally the development of adaptive immunity. The inflammatory response depends on a TLR-mediated MyD88-dependent cascade. However, there seems to exist additional cascades in TLR signaling. In the case of TLR4 signaling, an MyD88-independent pathway is now being characterized. In addition to the activation of innate immune responses, TLR-mediated signaling leads to suppression of the activity of innate immune cells, represented by "lipopolysaccharide (LPS) tolerance". Progress in elucidating the molecular mechanisms for LPS tolerance has been made through the analysis of TLR-mediated signaling pathways. Thus, the activity for innate immune responses is known to be finely regulated by TLRs. PMID- 11862003 TI - Molecular basis for innate immune recognition of microbial components. AB - Recognition of bacterial envelope constituents is one mechanism used by mammalian cells to initiate responses leading to bacterial killing, or, unfortunately, responses that also cause fatal septic shock. Many cell surface receptors by which these microbial components are recognized have been identified and characterized over the past a few years. In addition to CD14, which has been shown to be involved in the recognition of many microbial components, Toll-like receptors and MD-2 have been identified as factors playing a role in the receptor complexes of these components. Here we review the recent findings regarding the molecular basis for the recognition of microbial components. PMID- 11862004 TI - Non-specific interstitial pneumonia and Chlamydia pneumoniae infection. AB - Recently, the clinical features of non-specific interstitial pneumonia (NSIP) have been described. We hypothesize that recurrent infection caused by Chlamydia pneumoniae may play a role in the pathogenesis of NSIP. To prove this, we quantified serum IgA and IgG antibodies against C. pneumoniae using the enzyme linked-immunosorbent assay kit. The study included 15 patients diagnosed with NSIP, 20 patients with chronic obstructive pulmonary diseases (COPD) as disease group, and 27 control subjects. IgA antibody against C. pneumoniae was positive in 12 of 15 patients with NSIP, in 16 of 20 patients with COPD, and in 14 of 27 control subjects. IgG antibody against C. pneumoniae was positive in 14 of 15 patients with NSIP, in 17 of 20 patients with COPD, and in 16 of 27 control subjects. If the cut off value (mean +/- 2SD, index more than 3.0) was introduced, IgA and/or IgG antibodies against C. pneumoniae were positive in 8 of 15 patients with NSIP (53.3%), in 9 of 20 patients with COPD (45%), and in 2 of 27 control subjects (7.4%). These results suggest that infection of C. pneumoniae might play a role in the pathogenesis of NSIP. PMID- 11862005 TI - Characterization of phenotype-based pathogenic determinants of various Candida albicans strains in Jordan. AB - Sixty-six clinical isolates of Candida albicans representing 14 different strain types were tested for their phospholipase and proteinase activities in correlation with adherence to buccal epithelial cells (BECs) and lethality to mice. Variations in phospholipase and proteinase production as well as adherence to BECs were observed both among isolates of the same strain type and between isolates of different strain types. All isolates tested, irrespective of strain type, produced low levels of phopholipase (0.5 mm for strain -BCD- and 2.7 mm for strain ABC--) and acid proteinase (0.6 mm for strain A---E and 2.2 mm for strain -C--). A correlation was noted between adherence, phospholipase and proteinase production, and lethality to mice. C. albicans isolates, which adhered most strongly to BECs, exhibited higher levels of phospholipase and proteinase activities as well as higher pathogenicity. This was most evident in strain type -C--, which exhibited higher adherence ability (mean 717 +/- 21 yeasts/100 BEC), and proteinase activity (mean 2.2 mm), and relatively higher phospholipase activity (mean 2.4 mm) compared with those of other strains. Additionally, this type was more prevalent and showed significantly higher levels of tissue colonization in the liver, kidneys, and spleen compared with most other strain types in both subjects with healthy dentates and complete denture wearers. These results clearly demonstrate the significant role of phospholipase and proteinase activities on the adherence of C. albicans and their overall influence on the pathogenesis of Candida species. PMID- 11862006 TI - Utility of PCR in diagnosis of problematic cases of typhoid. AB - Typhoid is a global problem. Conventional diagnostic methods have limitations. The Widal test gives a high proportion of false positive results, and indiscriminate use of antibiotics has reduced the utility of blood culture. Consequently, these procedures are inadequate for diagnosing suspected cases of typhoid that do not present clear-cut symptoms. We previously showed that PCR based diagnosis of typhoid targeting the flagellin gene has unparalleled specificity. We assessed the utility of this method for diagnosis of problematic cases of typhoid. A comparative study of PCR, blood culture, and Widal test was carried out on 55 cases of suspected typhoid with fever for 3-30 days and possessing an ambiguous clinical picture. A control group comprised of 20 healthy persons was also included. The respective positive results by PCR, blood culture, and Widal test for these groups were 58.2 and 0%, 14.5 and 0%, and 52.7 and 45%. Sensitivity of PCR as compared with that of blood culture was significantly better. We concluded that PCR is much superior to conventional methods and, due to its high sensitivity and specificity, can be of great use for rapid and definitive diagnosis of problematic cases of typhoid. PMID- 11862007 TI - Molecular epidemiology of methicillin-resistant Staphylococcus aureus in a Tokyo hospital in 2001. PMID- 11862008 TI - A case of leptospirosis probably caused by drinking contaminated well-water after an earthquake. PMID- 11862009 TI - Identification of new parvovirus DNA sequence in swine sera from Myanmar. PMID- 11862010 TI - Skin eruption 8 days after a single dose of efavirenz-containing combination therapy. PMID- 11862011 TI - Enterohemorrhagic Escherichia coli O26 outbreak caused by contaminated natural water supplied by facility owned by local community. PMID- 11862012 TI - Isolation of echovirus type 13 from patients of aseptic meningitis. PMID- 11862013 TI - Necessity of two-stool sample test for sensitive detection of poliovirus. PMID- 11862015 TI - Local renin-angiotensin system in the pancreas: the significance of changes by chronic hypoxia and acute pancreatitis. AB - The circulating renin-angiotensin system (RAS) plays an important role in the maintenance of blood pressure and fluid homeostasis. Recently, there has been a shift of emphasis from the circulating RAS to the local RAS in the regulation of individual tissue functions via a paracrine and/or autocrine mechanism. In fact, a local RAS has been proposed to be present in an array of tissues including the brain, heart, kidney and gonads. Our previous studies have provided solid evidence that several key elements of the RAS, notably angiotensinogen and renin, are present in the rat pancreas. The data support the existence of an intrinsic RAS in the pancreas and this local RAS may be important for the exocrine/endocrine functions of the pancreas. Interestingly, such a pancreatic RAS has been demonstrated to be markedly activated by experimental rat models of chronic hypoxia and acute pancreatitis. The activation of the pancreatic RAS by chronic hypoxia and experimental pancreatitis could play a role in the physiology and pathophysiology of the pancreas. The significant changes of pancreatic RAS may have clinical relevance to acute pancreatitis and hypoxia-induced injury in the pancreas. PMID- 11862016 TI - The renin-angiotensin system: from the renal basis to an organ-specific subsystem in the pancreas. AB - Not only is the renin angiotensin system or its components found morphologically in many organs, it also exerts many different regulatory functions such as contributing to systemic homeostasis as well as to organ-specific regulation. The presence of the components of the renin angiotensin system in the pancreas was discovered only a few years ago. Physiological and pathophysiological stimuli were able to modify, in part, the gene expression and the occurrence of some of these components. Because of the important clinical significance of pancreatic diseases such as pancreatitis, research should follow every traces of the renin angiotensin system in the pancreas: impairment of microcirculation via hypoxia mediated up-regulation with the subsequent further deterioration of the oxygen supply seems to be the most obvious mechanism. There are many possible approaches to a better understanding of problems that are associated with diseases such as different kinds of pancreatitis; basic studies in animal models are oriented toward microcirculation, cellular function and the time course of modified gene expression after stimuli such as hypoxia; a clinical approach must reevaluate different correlations between clinical parameters of hypertension and those of pancreatic diseases. PMID- 11862018 TI - Association of free radicals and the tissue renin-angiotensin system: prospective effects of Rhodiola, a genus of Chinese herb, on hypoxia-induced pancreatic injury. AB - The renin-angiotensin system has long been recognized as crucial factor in the regulation of the systemic blood pressure and renal electrolyte homeostasis. Numerous studies have demonstrated the presence of a local renin-angiotensin system in a variety of organs. A recent study of the pancreatic renin-angiotensin system showed that chronic hypoxia significantly increased the mRNA expression for angiotensinogen II receptor subtypes AT1b and AT2. The activation of the renin-angiotensin system may play an important role in cellular pathophysiological processes. Angiotensin II enhances the formation of reactive oxygen species via the activation of xanthine oxidase or NAD(P)H oxidase. The reactive oxygen species can cause oxidative damage in the pancreas and other tissues either directly or indirectly via the formation of other radicals such as reactive nitrogen species. Rhodiola therapy may protect hypoxia-induced pancreatic injury in two ways. It prevents hypoxia-induced biological changes by increasing intracellular oxygen diffusion and efficiency of oxygen utilization. Alternatively, it reduces hypoxia-induced oxidative damage by its antioxidant activities. Additional experimental data are required to fully elucidate the mode of action of this herbal drug. PMID- 11862017 TI - Local renin-angiotensin system in the pancreas: the significance in acute pancreatitis. AB - Acute pancreatitis is a complex disease entity of which the pathogenesis is still not completely known. Research into the initiation and propagation of the diseases would hopefully help to design new treatment strategies for patients, especially those with severe acute pancreatitis. The novel observation of the activation of the local pancreatic renin-angiotensin system in experimental pancreatitis opens up new horizons for research regarding the pathogenesis of acute pancreatitis. PMID- 11862019 TI - The renin-angiotensin system in the endocrine pancreas. AB - Experimental data suggest that a tissue renin-angiotensin system is present in the pancreatic islets of several species, including man. However, the physiological role for this local renin-angiotensin system remains largely unknown. In vitro findings argue against a direct effect of angiotensin II on alpha- and beta-cells. In contrast, when the influence of angiotensin II on the pancreatic islets has been evaluated in the presence of an intact vascular system either in vivo or in the perfused pancreas, a suppression of insulin release has been observed, also in man. These discrepancies may be explained by the profound effects of the renin-angiotensin system on pancreatic islet blood perfusion. Alterations in the systemic renin-angiotensin system and an increased vascular sensitivity for its components have been observed in diabetes mellitus and hypertension. Whether changes occur also in the pancreatic islet renin angiotensin system during these conditions remains unknown. Future research may help to provide an answer to this question, and to elucidate to what extent the renin-angiotensin system may contribute to beta-cell dysfunction in these diseases. PMID- 11862020 TI - The renin-angiotensin system and the exocrine pancreas. AB - An accumulating body of evidence strongly indicates a local tissue renin angiotensin system in the pancreas of a various species. In contrast to the majority of tissues that primarily express the angiotensin type 1 (AT1) receptor, the pancreas is one of the few tissues that contain a significant proportion of the AT2 subtype. Moreover, our findings indicate a greater distribution angiotensin II binding sites in the exocrine pancreas. Although the physiological aspects of a local pancreatic renin-angiotensin system remain largely unexplored, recent studies in our laboratory utilizing an acinar cell model demonstrate both functional AT1 and AT2 receptors. Indeed, we show that the AR42J cell line expresses all components of an angiotensin system including the mRNA for renin, angiotensinogen, angiotensin converting enzyme (ACE), AT1a, AT1b and AT2 receptors. Thus, these cells may be of particular value to study the interplay of the AT1 and AT2 receptors to regulate cell growth and potentially exocrine function. PMID- 11862021 TI - The pancreatic renin-angiotensin system: does it play a role in endocrine oncology? AB - The characterization of a local renin-angiotensin system in the pancreas has attracted much attention because of its potential clinical applications. A pancreatic renin-angiotensin system may be present in humans and may interact with islet cells. Nevertheless, our knowledge of the renin-angiotensin system in the human pancreas is still in its infancy, especially in the field of endocrine oncology. Much of our knowledge stems from the study of the pancreas and pancreatic endocrine tumors of rodents. Thus, the direction of future research should be based on in-depth and collaborative efforts between researchers in the various disciplines in order to apply the newly acquired scientific knowledge to the patient. PMID- 11862022 TI - Possible involvement of the local renin-angiotensin system in exocrine pancreas responses to food components. AB - The functioning of the exocrine and endocrine pancreas is strictly co-ordinated through an interdependent array of neural and endocrine, paracrine and autocrine hormonal factors. The responses of the exocrine pancreas to food are primarily initiated via hormones secreted by neuroendocrine cells in the gut. No role for the pancreatic renin-angiotensin system in these mechanisms has so far been established. However, because of its distribution throughout the pancreas, the renin-angiotensin system could have a function in fine-tuning of secretory responses or in integrating some of the actions of the endocrine and exocrine pancreas. In the normal diet, we are exposed to an array of bioactive (lectins, protease inhibitors, hormone-mimics, tannins, etc). Some can profoundly alter pancreas metabolism both in a beneficial or detrimental manner. Others could have beneficial effects on the pancreas renin-angiotensin system. The effects of these compounds need to be evaluated. PMID- 11862023 TI - A critical appraisal of the intrinsic pancreatic angiotensin-generating system. AB - The pancreas is a relative newcomer to the stable of tissues with an intrinsic angiotensin-generating system. The involvement of this system in pancreatic activity will be dependent on the angiotensin-generating paths present in the pancreas and their precise cellular location. Thus far, renin, angiotensin converting enzyme (ACE), angiotensin II and AT1 and AT2 receptors have been found. These are components of the "classical" renin-angiotensin system. But there is uncertainty as to their location and site of action. Furthermore, it is not known which, if any, alternative enzymes to renin and ACE are present, which angiotensins in addition to angiotensin II are generated and whether or not there are receptors to angiotensin IV and angiotensin-(1-7). Future research should focus on these aspects in order to provide a mechanistic basis to pancreatic physiological functions and to pathological conditions of clinical relevance. PMID- 11862024 TI - Renin-angiotensin system in the pancreas: from the basic research to the bedside. Conclusive remarks. PMID- 11862025 TI - Epidemiological investigation of local complications after cosmetic breast implant surgery in Denmark. AB - Concern has been raised recently regarding the absence of information on the occurrence and severity of local complications after cosmetic breast implantation. The authors evaluated the occurrence of local complications in a large epidemiological retrospective cohort study of women with cosmetic breast implants in Denmark. All women with breast implants were identified from the files of two private clinics of plastic surgery in Denmark. Information on implant characteristics, surgical procedure, as well as short- and long-term complications was obtained through medical record review. Patient characteristics were obtained through a self-administered questionnaire. A total of 754 women (1,572 implants) had at least one cosmetic implantation performed at the study clinics. Average age at implantation was 32 years. Implant types included silicone double lumen with textured surface, 31.2%; silicone single lumen with textured surface, 27.8%; silicone single lumen with smooth surface, 24.5%; silicone double lumen with smooth surface, 0.8%; and other or missing information, 15.7%. Average implant size was 247 ml (range, 110-630 ml). Placement was submuscular for 91.3% of implants, subglandular for 2.6%, and 6.1% had no available information. Overall, 77.8% of implantations were not followed by complications, 17.8% were followed by one complication, 3.6% were followed by two complications, and 0.8% were followed by three or more complications. In 94.6% of implantations, no additional hospitalizations were recorded as a result of complications. Forty-seven of 57 explantations/reimplantations were secondary to postoperative complications. General complications such as hematoma and infection were rare, occurring in 2.3% and 2.0% of implantations respectively. Capsular contracture remains the most common complication, occurring in 11.4% of implantations. In this investigation, among the first epidemiological studies of local complications, the authors found cosmetic breast implant surgery to be associated with a low frequency of normal surgical complications such as infection, hematoma, and wound dehiscence. Most complications were mild and did not lead to additional hospitalization. The complication that led most frequently to the need for additional surgery/hospitalization was capsular contracture. Kjoller K, Holmich LR, Jacobsen PH, Friis S, Fryzek J, McLaughlin JK, Lipworth L, Henriksen TF, Jorgensen S, Bittmann S, Olsen JH. Epidemiological investigation of local complications after cosmetic breast implant surgery in Denmark. PMID- 11862026 TI - Health outcomes in offspring of Danish mothers with cosmetic breast implants. AB - To assess whether maternal breast implants are related to adverse health outcomes in offspring, the authors conducted a retrospective cohort study of esophageal disorders, rheumatic disease, and congenital malformations among 2,854 children born to women with breast implants and among 5,805 children born to a comparison group of women who underwent breast reduction or other plastic surgery. Rates were calculated using both hospitalization and outpatient data. Significantly higher rates of esophageal disorders were observed (O) than were expected (E) for children born before (O/E, 2.0; 95% CI, 1.3-2.8) but not after (O/E, 1.3; 95% CI, 0.5-2.9) maternal breast implant surgery. Risk of rheumatic disease was similar among children born before (O/E, 1.4; 95% CI, 0.7-2.6) and after (O/E, 1.4; 95% CI, 0.2-5.0) maternal breast implant surgery. A marginally significant excess of congenital malformations of the digestive organs was observed among children born after maternal implant surgery (O/E, 1.8; 95% CI, 1.0-3.1), with a similar finding among children born to women in the comparison group (O/E, 1.9; 95% CI, 1.4-2.4). The risk of malformations overall was not statistically significantly higher than expected among children born after maternal breast implant surgery. The elevated risks of adverse health outcomes appear unrelated to breast implants per se, because similar findings were observed among children born both before and after the mother's implant surgery, as well as among children born to mothers in the comparison cohort. PMID- 11862027 TI - Laser blepharoplasty in Asians. AB - Traditional blepharoplasty removes periorbital wrinkles by cutting and stretching the skin. However, this method has a substantial risk of producing ectropion or scleral show. In addition, fine periocular wrinkles may persist because this method does not change skin texture. The pulsed CO2 laser has recently become a primary surgical tool in treating aging eyelids. Periorbital wrinkles vary in depth not only from person to person, but also among different races. Compared with whites, most Asians have a thicker dermis, so more laser passes and a higher power may be required to remove periorbital wrinkles, but concerns about hyperpigmentation and prolonged erythema have limited its use on Asian skin. In this study, 346 patients underwent laser blepharoplasty at the Korea University Medical Center and at Dr Choi's Aesthetic Clinic. They were followed for 12 months on average from September 1995 to September 1999. The CO2 laser was used in resurfacing periorbital wrinkles, transcutaneous skin excision, and transconjunctival blepharoplasty, including fat removal. The authors assessed the benefit of using the UltraPulse CO2 laser in Asian blepharoplasty. They found that 291 patients (84%) had good to excellent results. The incidence of side effects was very low. Prolonged erythema occurred in 19 patients (5%) and hyperpigmentation occurred in 35 patients (10%), but the erythema disappeared spontaneously within 2 months and the hyperpigmentation could be managed readily by the topical use of retinoids and hydroquinone cream. Therefore, the authors conclude that postoperative hyperpigmentation is no longer a problem limiting laser resurfacing in Asian blepharoplasty. The UltraPulse CO2 laser is a safe and effective rejuvenation method for treating aging eyelids in Asians. PMID- 11862028 TI - Thin anterolateral thigh free flap. AB - The need for a thin flap has increased for contour or coverage of the shallow defects caused by trauma, tumor ablative surgery, or defects created after the release of contractures. The authors describe their experience with the use of an extremely thin anterolateral thigh free flap for covering such defects in a series of 12 patients. Extreme thinning of the flap (4-5 mm) was achieved by removal of deep fascia and subcutaneous fat except for a 3- to 4-cm area around the entry of the perforator into the flap. Subdermal fat and immediate underlying superficial veins should be preserved during the thinning procedure for venous drainage of the flap. Their clinical experiences with 12 patients indicate that an extremely thin, long flap can survive on a single perforator. PMID- 11862029 TI - Soft-tissue reconstruction of the foot with distally based neurocutaneous flaps in diabetic patients. AB - Soft-tissue coverage of the foot in diabetic patients is often a difficult problem to undertake. The aim of this study was to evaluate the efficiency and safety of distally based neurocutaneous flaps for foot reconstruction in diabetic patients. The authors describe their experience with a series of 10 diabetic patients in whom reconstruction of defects of the foot (range, 6 x 11-10 x 12 cm) were performed using distally based sural and saphenous neurocutaneous flaps. In 9 patients the flap survived completely and in only 1 patient was superficial marginal necrosis of the flap observed. Partial skin graft loss at the donor site occurred in 1 patient, but no additional surgical revision was needed and healing occurred by secondary intention. In these diabetic patients, defects were reconstructed successfully using neurocutaneous flaps, and good results were achieved. PMID- 11862030 TI - Complications and removal rates of miniplates and screws used for maxillofacial fractures. AB - Complications of miniplates and screws used for maxillofacial fractures were analyzed, and complications were evaluated in relation to fracture site. Motor vehicle accidents were the cause of all fractures in this study. During the last 7 years (1994-2001), noncompressive titanium miniplates and screws were used for stabilization of maxillofacial fractures. In 66 patients, 87 fracture sites were stabilized using 296 miniplates and 1,184 screws. The mean age of the patients was 31 years (age range, 6-64 years). The percentage of male patients was 77% and the percentage of female patients was 23%. Miniplates and screws were used in 6 patients (10%) who were younger than 15 years of age at the time of the surgery. The follow-up period ranged between 3 months and 7 years. The overall miniplate and screw removal rate was 7%. The rates of removal according to the fracture site are as follows: mandible, 4.4%; zygomaticofrontal junction, 1.4%; inferior orbital rim, 0.7%; maxilla, 0.3%; and frontal sinus wall, 0.3%. Removal causes were infection, 2%; extrusion, 1.7%; visibility, 1.4%; pain, 1%; malunion, 0.7%; and miniplate fracture, 0.3%. The minimum time period between insertion and removal was 3 months and the maximum period was 14 months. Infection and extrusion were the main complications for removal of miniplates and screws from the mandible, whereas miniplates and screws were removed from the zygoma because of visibility (zygomaticofrontal region) under the skin in the vast majority of the patients. The maxilla was the least operated region for miniplate and screw removal. In all patients in this study, the preoperative physical symptoms were relieved after miniplate and screw removal. Miniplates and screws are very useful tools in maxillofacial fracture management, but sometimes they have to be removed. In the authors' series, the removal rate was 7%, and this rate can vary with the severity of the trauma and location of the fracture. PMID- 11862031 TI - Prevalence of smoking, obesity, diabetes mellitus, and thyroid disease in patients with carpal tunnel syndrome. AB - Carpal tunnel syndrome (CTS) has been associated frequently with physical factors and personal factors including smoking, obesity, diabetes mellitus, and hypothyroidism. The purpose of this retrospective case-control study was to evaluate the prevalence of personal factors in patients with CTS compared with a normal control group. The charts of 514 patients who underwent a carpal tunnel release procedure and 100 control subjects were examined for history of smoking, obesity, diabetes, and hypothyroidism. Compared with the control subjects, there were significantly less CTS patients who smoked (p < 0.001; odds ratio, 0.17). However, there were more CTS patients than control subjects who were obese (p = 0.02; odds ratio, 1.77), had diabetes (p = 0.03; odds ratio, 3.02), and had hypothyroidism (p = 0.02; odds ratio, 3.70). These results support previous reports that CTS is multifactorial, with such factors as obesity, hypothyroidism, and diabetes but not smoking to be more prevalent in this group of CTS patients. PMID- 11862032 TI - Patient self-reported outcome after ulnar nerve transposition. AB - Subjective outcome was assessed in 100 patients with cubital tunnel syndrome at least 2 years after anterior transmuscular transposition of the ulnar nerve. The mean time from surgery was 37 months (standard deviation, 10 months). The patient sample of 119 operated cases included 81 unilateral procedures and 19 patients had bilateral procedures. The mean time between surgeries was 7 months. Sixty-one patients who underwent unilateral procedures reported improvement, and there was no difference in 13 unilateral cases. Thirteen patients who underwent bilateral procedures reported improvement in both arms. Postoperatively symptoms were significantly less than those preoperatively: pain, p = 0.001; cold sensitivity, p = 0.001; tingling, p = 0.05; and numbness p = 0.08. In 51 cases, patients reported normal sensation and in 45 cases patients reported normal strength. Reported outcome was significantly better in nonsmokers than in smokers (p = 0.02) but was not significantly different in patients with workers' compensation/litigation (p = 0.51), obesity (p = 0.16), abnormal preoperative nerve conduction studies (p = 0.21), concomitant carpal tunnel syndrome (p = 0.95), or brachial plexus nerve compression (p = 0.35). PMID- 11862033 TI - Epineural sleeve neurorrhaphy: surgical technique and functional results--a preliminary report. AB - This study was conducted to evaluate the effect of epineural sleeve neurorrhaphy on peripheral nerve regeneration. A total of 12 Lewis rats were divided in two groups of 6 rats each. In group 1, the rat sciatic nerve was transected and repaired using the conventional epineural technique with four sutures. In group 2, the epineural sleeve technique was introduced with two sutures only. Functional recovery was evaluated at 1, 2, 4, 8, and 12 weeks by walking track analysis (sciatic function index [SFI]), mean limb circumference ratio, and severity of toe contracture. Although the SFI at 12 weeks revealed no difference between the two groups (+/-88.39 +/- 10.75 conventional group, +/-77.35 +/- 17.06 epineural sleeve group), significant differences in SFIs were detected at 4 and 8 weeks, with better functional recovery in group 2 rats (4 weeks: 125.92 +/- 22.73 conventional group, +/-99.17 +/- 5.45 epineural sleeve group; 8 weeks: +/-96.65 +/- 4.73 conventional group, +/-72.82 +/- 17.11 epineural group; p < 0.05 for both time points). Mean limb circumference ratio was not significant at all time points. At 12 weeks, all animals in the conventional nerve repair group developed severe toe contractures whereas only 2 animals in epineural sleeve repair group had contracture (p < 0.05). In this study, the epineural sleeve technique demonstrated a faster functional recovery when compared with the conventional technique, as confirmed by SFI and toe contracture grading. PMID- 11862034 TI - Posterior thigh perforator-based flap: a new experimental model in rats. AB - Animal research has added a great deal of understanding to flap hemodynamics. The rat is the most commonly used animal in flap research, and various flap models have been devised. In the current study the authors developed a single-perforator based flap model in rats. In 30 rats, anatomic dissection and flap elevation based on a single musculocutaneous perforator artery arising from the biceps femoris muscle were performed. The vascular basis of this new flap was cleared by anatomic dissection of the posterior thigh in 6 rats. The survival pattern of the proposed flap was investigated in three groups of rats. Each group consisted of 8 rats and had different flap dimensions. In the first group the flap was located in the posterior thigh and was 3 x 2 cm. All flaps survived in this group at the end of the first week. In the second group the same flap was extended to the gluteal region and was 3 x 4 cm. Again, all flaps survived. In the last group an oversized flap (3 x 12 cm) was planned from the posterior thigh to the scapular region based on the same perforator artery. In this group only 61 +/- 7.2% (mean +/- standard deviation) of the flap survived at the end of the first week. Microangiography was performed in each group and the vascular architecture of the pedicle (perforator artery) was seen. This new posterior thigh perforator-based flap model is simple and reliable with a constant survival pattern. Thus, it could be used in studies investigating the physiological and pathophysiological changes of perforator-based flaps. PMID- 11862036 TI - Regeneration of neomucosa using cell-seeded collagen-GAG matrices in athymic mice. AB - Tissue engineering of oral mucosa could allow improved reconstructive options for intraoral tissue defects. Porous collagen-glycosaminoglycan (CG) matrices coated with a silicone elastomer were seeded by centrifugation with cultured human oral mucosal epithelial cells (strain OKG4 gingival keratinocytes) at a density of 250,000 cells per square centimeter. Full-thickness dorsal wounds (1.5 x 1.5 cm) were created surgically on each athymic mouse and were treated with either a seeded matrix or an unseeded matrix, or they were left open as a control. The CG matrices reduced the degree of wound contraction at day 14 compared with open wounds. The epithelial thickness of seeded matrices at day 14 was significantly greater (p = 0.0001) than that of unseeded matrices. Seeded matrices had more rapid degradation at 14 days compared with unseeded matrices. Human oral mucosal cells seeded into CG matrices contribute to form a stratified and differentiated epithelial layer during revascularization, cellular infiltration, and degradation of the CG matrix. PMID- 11862035 TI - Expression of cyclooxygenase-2 in the periprosthetic capsule surrounding a silicone shell implant in the rat. AB - Prosthetic breast implants are used frequently for both aesthetic and reconstructive purposes. Histologically, the normal tissue response to silicone implants typically involves an inflammatory infiltrate that consists initially of macrophages, and later consists of fibroblasts, myofibroblasts, and lymphocytes. To characterize further the nature of the inflammatory response to silicone breast implants, the authors evaluated the expression of the enzyme cyclooxygenase-2 (COX-2) by the tissues and cells of the breast capsule after silicone augmentation in an animal model. COX-2 is an inducible enzyme that is expressed primarily in response to inflammatory stimuli and mediates the production of prostaglandins that support the inflammatory process. Fifty-nine animals at five time points were evaluated. Specifically, on days 4, 7, 14, 28, and 80 the authors identified cytoplasmic COX-2 expression in each of the three cell types of interest, including endothelial cells, macrophages, and fibroblasts. Selective COX-2 inhibiting agents may be an appropriate pharmacological intervention for modulating the formation of periprosthetic capsules. PMID- 11862037 TI - Effect of long-term cyclosporine administration on muscle flap hemodynamics. AB - Long-term use of cyclosporine A (CsA) is associated with deleterious effects such as nephrotoxicity and hypertension as a result of its toxicity on microvasculature. These effects raise the possibility that long-term CsA use harms the microvasculature of the skeletal muscle tissue. An experimental study was conducted to investigate the effect of chronic systemic cyclosporine administration on the microvasculature of the cremaster flap model in the rat. Thirty male Sprague-Dawley rats were divided into five groups of 6 animals each. The control group received no treatment. The four CsA treatment groups received a daily subcutaneous injection of 2 mg per kilogram, 4 mg per kilogram, 8 mg per kilogram, and 16 mg per kilogram of cyclosporine for 6 weeks before surgery. The effect of long-term CsA administration on cremaster flap microcirculation was evaluated in vivo using an intravital microscopy system. Hemodynamic parameters of the cremaster muscle flap such as vessel diameter, red blood cell velocity, capillary density, leukocyte-endothelial interaction, and microvascular permeability were measured. There was no significant (p > 0.05) difference in vessel diameter in all groups. There was a significant increase in the number of adherent leukocytes in the 8-mg and 16-mg CsA group compared with the control (12 +/- 4.8 leukocytes per 100 microm and 12 +/- 4.5 leukocytes per 100 microm vs. 6 +/- 4.3 leukocytes per 100 microm; p < 0.05). Microvascular permeability indices increased significantly at 0 and 30 minutes after fluorescent isothiocyanate albumin injection in the 8-mg and 16-mg CsA groups compared with the control (0 minutes: 0.6 +/- 0.2% and 0.5 +/- 0.1% vs. 0.4 +/- 0.05%; 30 minutes: 0.8 +/- 0.2% and 0.7 +/- 0.1% vs. 0.5 +/- 0.04%; p < 0.05). Histologically, the cremaster muscle flaps in all cyclosporine groups showed evidence of interstitial inflammation and venous vasculitis. In the 8-mg and 16-mg CsA groups there was also focal muscle injury. The toxic effect of CsA on the microvascular tree of a muscle flap was demonstrated by the increased permeability index in vivo, and the moderate venulitis and focal muscle injury histologically. Systemic CsA administration seems to have minimal impact on the viability of the muscle flaps, which was confirmed by preserved capillary function and muscle flap perfusion. These data suggest that there is a minimal risk in undertaking a pedicled muscle flap transfer procedure using a CsA immunosuppressive protocol. PMID- 11862038 TI - Case of constriction band syndrome with annular epidermal cyst. AB - Both intrinsic and extrinsic theories have been proposed to explain the etiology of congenital constriction band syndrome. The authors report a case of construction band syndrome in which an epidermal cyst was involved. It was located along the annular band of the affected finger and formed a complete ring. This case suggests that the epidermal cyst was produced by circumferential compression, advocating the extrinsic theory for this syndrome. PMID- 11862039 TI - Eyebrow island flap for reconstruction of a partial eyebrow defect. AB - A diverse variety of methods for reconstructing eyebrow defects has been described previously, and each procedure has inherent advantages and disadvantages. The authors present a case of reconstruction of a partial eyebrow defect using two eyebrow island flaps, which are modifications of the subcutaneous pedicle flap. PMID- 11862040 TI - Rare complication of breast cancer irradiation: postirradiation osteosarcoma. AB - Radiation-induced osteosarcoma is a rare complication of radiation therapy for breast cancer. The authors present a 60-year-old patient in whom osteosarcoma of the chest wall developed 5 years after modified radical mastectomy and radiation therapy for breast cancer. One year after resection of the chest osteosarcoma, metastasis to the contralateral axillary lymph nodes developed and these were removed. Radiation-induced osteosarcoma is difficult to treat and has a poor prognosis, thus early diagnosis is necessary for optimal treatment. PMID- 11862041 TI - Digitization of photographic slides: simple, effective, fast, and inexpensive method. AB - The technological evolution has changed multiple areas of plastic surgery, including photography. The photograph is one of the instruments used most by the plastic surgeon, and it cannot be eliminated by technological changes. The principal change in photography is that images can be scanned through digital cameras instead of slides. Despite the multiple advantages that digital photography represents, many surgeons are resisting the change. One of the main reasons for this resistance is the large quantity of photographic slides that need to be digitized to be used at scientific conferences as well as in publications. The methods and existing techniques for digitizing slides are costly and time-consuming, and there is risk for loss of definition and image brightness. The authors present a simple, effective, fast, and inexpensive method for digitizing slides. This method has been validated by various plastic surgeons and is effective for use in multimedia presentations and for paper printouts with publication quality. PMID- 11862043 TI - Lord of our ring. PMID- 11862042 TI - Invited discussion of surgical repair for congenital macrostomia: vermilion square flap method. PMID- 11862044 TI - A breast-to-lip filler. PMID- 11862045 TI - Multiple aesthetic surgical procedures under conscious sedation and local anesthesia. PMID- 11862046 TI - Epidermoid carcinoma arising in a giant condyloma acuminata of 20 years' duration. PMID- 11862047 TI - Comparison of three methods of oral reconstruction for quality-of-life assessment. PMID- 11862048 TI - Coverage of exposed femoral vascular prosthesis using a rectus abdominis muscle flap. PMID- 11862050 TI - Cystic clear cell hidradenoma of the thumb: case report of a rare hand tumor. PMID- 11862051 TI - Flipper hand. PMID- 11862052 TI - Whistling face syndrome associated with camptodactyly. PMID- 11862049 TI - Osteochondroma arising from the mandibular symphysis. PMID- 11862053 TI - Unknown artifact. PMID- 11862054 TI - Recommending medications during pregnancy: an evidence based approach. PMID- 11862055 TI - Drug effects on the fetus and breast-fed infant. PMID- 11862056 TI - Hypertension therapy during pregnancy. PMID- 11862057 TI - Corticosteroids and fetal pulmonary maturity. PMID- 11862058 TI - Immunization during pregnancy. PMID- 11862059 TI - Antibiotic selection in obstetrics: making cost-effective choices. PMID- 11862060 TI - Drug abuse during pregnancy. PMID- 11862061 TI - Over-the-counter products: nonprescription medications, nutraceuticals, and herbal agents. PMID- 11862063 TI - Cervical ripening agents and uterine stimulants. PMID- 11862062 TI - Pharmacologic inhibition of preterm labor. PMID- 11862064 TI - Pharmacologic therapy for peripartum emergencies. PMID- 11862065 TI - Pharmacologic therapy for medical disorders during pregnancy. PMID- 11862067 TI - Management of diabetes in pregnancy. PMID- 11862066 TI - Chemotherapy and pregnancy. PMID- 11862070 TI - The overactive bladder: prevalence and effects on quality of life. PMID- 11862071 TI - Pathophysiology of the overactive bladder. PMID- 11862072 TI - Evaluation and diagnosis of the overactive bladder. PMID- 11862073 TI - Pharmacotherapy of the overactive bladder and advances in drug delivery. PMID- 11862074 TI - Neuromodulation techniques for the treatment of the overactive bladder. PMID- 11862075 TI - Gynecologic manifestations of interstitial cystitis. PMID- 11862076 TI - Interstitial cystitis: epidemiology and clinical presentation. PMID- 11862077 TI - Diagnosis of the painful bladder syndrome: current approaches to diagnosis. PMID- 11862078 TI - Interstitial cystitis: a pathophysiology and treatment update. PMID- 11862079 TI - Pelvic floor muscle re-education treatment of the overactive bladder and painful bladder syndrome. PMID- 11862081 TI - The clinical efficacy of silicone punctal plug therapy. AB - PURPOSE: To evaluate the clinical efficacy and safety of silicone punctal plugs in various ocular surface disorders. PATIENTS AND METHODS: Medical records of patients who underwent silicone punctal plug insertion from January 1996 to November 2000 were retrospectively reviewed. The clinical data collected included sex and age of the patients, indications for plug insertion, symptoms, corneal fluorescein staining and frequency of lubricant use at baseline and at 4 +/- 2 weeks after the insertion, location (upper versus lower punctum), initial versus replacement plug, spontaneous extrusion or removal of the plug, and percentage of complications and punctum cauterization. RESULTS: There were 203 eyes of 153 patients (126 women and 27 men) in the study group. The total number of silicone punctal plug procedures was 312. Dry eye syndrome was the most common indication for punctal plug treatment (127 eyes, 62.5%), followed by epitheliopathy after penetrating keratoplasty (32 eyes, 15.8%). The symptoms improved in 150 (73.9%) of 203 eyes at 4 +/- 2 weeks' follow-up. The mean score of the fluorescein staining of the cornea was reduced from 2.7 +/- 0.1 to 1.3 +/- 0.1 with silicone punctal plug treatment at 4 +/- 2 weeks' follow-up. This reduction was statistically significant (p< 0.01). The overall success rate at 4 +/- 2 weeks follow up was 76.8%. The frequency of the lubricant use was significantly decreased with punctal plug treatment (p< 0.001). Considering all plugs, the estimated probability of plug retention was 49.4% with 85.1 +/- 7.3 weeks as the mean survival time. There was no statistical difference in plug retention rates between lower (49.6%) and upper (48.7%) plugs (p= 0.15). Retention rate of first plugs (53.5%) was significantly higher than that of second plugs (32.0%) (p= 0.05). The spontaneous total extrusion of silicone punctal plug was the most common complication in our study (158 plugs, 50.7%). The plug was removed in 14 eyes (14 plugs, 6.9%) because of epiphora in 11 eyes and conjunctival erosion in three eyes. The percentage of punctum cauterization performed in our population was 10.8%. CONCLUSIONS: Punctal plug insertion is a simple, effective, safe, and reversible method to treat aqueous tear deficiency and other ocular surface diseases, such as epitheliopathy after penetrating keratoplasty, superior limbic keratoconjunctivitis, neurotrophic keratitis, recurrent corneal erosions, and toxic epitheliopathy not controlled by lubrication with preservative-free tears. However, spontaneous extrusion of silicone plug occurs commonly. Careful choosing of the optimal plug size and close follow-up are recommended to ensure the success of the treatment. PMID- 11862082 TI - Topographic analysis of corneal regularity after penetrating keratoplasty. AB - PURPOSE: To evaluate the accuracy of the Holladay Diagnostic Summary of the EyeSys Corneal Analysis System in predicting the corneal visual acuity potential in patients who have undergone penetrating keratoplasty (PKP). METHODS: Astigmatism patterns, refractive and topographic astigmatism, best spectacle corrected visual acuity (BSCVA), and hard contact lens (HCL) visual acuity of 54 patients were analyzed 3 months after PKP and compared with the Potential Corneal Acuity (PCA) value predicted by the Holladay Diagnostic Summary. RESULTS: Qualitative patterns of astigmatism (p = 0.01) and refractive (p = 0.002) and topographic (p = 0.0002) astigmatism were significantly correlated with PCA values. Using HCL visual acuities to correct the BSCVA (HCL-corrected BSCVA) for noncorneal causes of reduced vision, we found that the PCA values of 48.1% of the patients were within one line of the HCL-corrected BSCVA; 81.5% were within two lines; and 93.0% were within three lines. CONCLUSION: The Holladay PCA measurement may be useful in the postoperative evaluation of the optical quality of the central corneal surface in patients who have undergone PKP. PMID- 11862083 TI - Indications for penetrating keratoplasty and associated procedures, 1996-2000. AB - PURPOSE: To identify current indications and trends in indications for penetrating keratoplasty and associated procedures. METHODS: Retrospective chart review of all patients who underwent penetrating keratoplasty at Wills Eye Hospital from January 1, 1996 to December 31, 2000. RESULTS: A total of 1,529 corneal transplants were performed during this 5-year period. The leading indications for penetrating keratoplasty were pseudophakic bullous keratopathy (PBK) in 416 (27.2%) cases, followed by regraft in 276 (18.1%), keratoconus in 236 (15.4%), and Fuchs endothelial dystrophy in 233 (15.2%) cases. Of the 416 PBK cases, 271 (65.1%) were associated with posterior chamber intraocular lenses (PC IOLs), 141 (33.9%) with anterior chamber intraocular lenses (AC IOLs), and four (1%) with iris-fixated lenses. Of these 141 AC IOLs, 107 (75.9%) were closed-loop and 34 (24.1%) were open-loop. Of the 416 PBK cases, in 281 (67.5%) cases, the IOL was not exchanged; in 127 (30.5%) cases, the IOL was exchanged; in six (1.5%) cases, the IOL was repositioned; and in two (0.5%) cases, the IOL was removed at the time of surgery. Of the 271 PC IOLs, 248 (91.5%) were not exchanged and 19 (7.1%) were exchanged. Of the 141 AC IOLs, 33 (23.4%) were not exchanged and 104 (73.7%) were exchanged (p = 0.001). CONCLUSIONS: Pseudophakic bullous keratopathy remains the leading indication for corneal transplantation at our institution followed by regraft. The percentage of PBK cases associated with PC IOLs has increased significantly, whereas the percentage associated with AC IOLs has decreased. The frequency of regraft has also increased significantly. PMID- 11862084 TI - Indications for corneal transplantation in New Zealand: 1991-1999. AB - PURPOSE: To identify the indications for keratoplasty in patients supplied with donor tissue through the New Zealand National Eye Bank. METHODS: Analysis of penetrating and lamellar keratoplasty data collected by the New Zealand National Eye Bank, Auckland, from 1991 to 1999. RESULTS: In this 9-year period, donor material was supplied for 1370 corneal grafts; 1308 for penetrating keratoplasty, 26 for lamellar keratoplasty, and 36 for unspecified grafts. This accounts for a minimum of 85% of the penetrating keratoplasties performed in New Zealand from 1991 to 1999. The leading indications for penetrating keratoplasty were keratoconus (45.6%), pseudophakic or aphakic corneal edema (17.9%), regraft (8.7%), viral keratitis (7.3%), and trauma (5.5%). The average age of patients was 47.5 years (SD = 22.6) and age distribution was bimodal, with peaks in the 3rd and 8th decades. Keratoconus, regraft, and trauma were significantly more common as indicators for penetrating keratoplasty in male patients than female patients; however, pseudophakic or aphakic corneal edema was more common in female patients. CONCLUSION: The majority of transplantation surgery in New Zealand is performed using corneal tissue from the New Zealand National Eye Bank. In this representative study, keratoconus is the leading indicator for penetrating keratoplasty in New Zealand, accounting for a higher proportion than in any other published literature. The other indications, age distribution and gender differences correlate with previous reports. These findings suggest that keratoconus leading to transplantation may have increased prevalence in New Zealand. PMID- 11862085 TI - Assessment of applanation tonometry after hyperopic laser in situ keratomileusis. AB - PURPOSE: To determine the reliability and identify pitfalls in intraocular pressure measurement by Goldmann applanation tonometry after hyperopic laser in situ keratomileusis (LASIK). METHODS: Prospective non-masked case series at University of Valencia, Faculty of Medicine and Rahhal Ophthalmology Clinic, Valencia, Spain. One hundred three patients (103 eyes) treated with hyperopic LASIK were evaluated. The main treatment was hyperopic LASIK using a microkeratome Chiron Hansatome (Chiron Vision Corp, Claremont, CA) and the excimer laser Chiron Technolas 217-C LASIK (Chiron Technolas GmbH, Dornack; Germany). Central Goldmann applanation tonometric readings before surgery and 1, 3, and 6 months after surgery were analyzed. RESULTS: After hyperopic LASIK, a significant decrease in intraocular pressure was observed in the postoperative controls. In the low hyperopia patients (range: 1.00-3.00 D) a decrease of 2.43 mmHg was observed at the 6-month follow-up (p < 0.001). In the moderate hyperopia group (range: 3.25-6.00 D) a decrease of 2.05 mmHg was observed at the 6-month follow-up (p < 0.001). There were no significant differences between low and moderate hyperopia (p = 0.812). There was no statistically significant correlation between the magnitude of decrease in tonometry and gender, age, treated spherical equivalent, pachymetry, and anterior chamber depth (ACD). CONCLUSIONS: This clinical study displays that postoperative tonometry after hyperopic LASIK was significantly lower than the preoperative, hence modifying the reliability of Goldmann tonometry, and causing an intraocular pressure underestimation. A correcting factor should be applied when using applanation tonometry to measure postoperative intraocular pressure in patients who have undergone hyperopic LASIK. PMID- 11862086 TI - Topical cyclosporin A in the management of therapeutic keratoplasty for mycotic keratitis. AB - PURPOSE: To investigate the efficacy of topical cyclosporine A 0.5% as a substitute for corticosteroids in the management of therapeutic keratoplasties for mycotic keratitis. METHODS: Prospective, nonrandomized interventional case series. Three patients with culture-proven mycotic keratitis underwent therapeutic keratoplasties. All were treated with topical cyclosporine A 0.5% postoperatively as a primary or an adjunctive therapy for prevention of allograft rejection. The patients were followed up from 15 to 42 months for maintenance of graft clarity and best-corrected visual acuity. RESULTS: Two of three patients maintained clear grafts while using topical cyclosporine A 0.5% twice daily with best-corrected visual acuity of 20/40 and 20/50. One patient developed an opacified graft secondary to preexisting ocular surface disease. CONCLUSIONS: Topical cyclosporine A 0.5% may be a useful adjunct in the management of therapeutic keratoplasties for mycotic keratitis. PMID- 11862087 TI - N-acetylcysteine in chronic blepharitis. AB - PURPOSE: To investigate the effects of N-acetylcysteine (NAC) in chronic posterior blepharitis. METHODS: This was a prospective randomized, controlled study that included 79 eyes of 40 patients with chronic posterior blepharitis. Routine ophthalmologic examination, Schirmer-1 test, fluorescein break-up time (FBUT), and mucous fern tests were carried out during the first visit of all patients. A topical steroid, topical antibiotic, and artificial tears were started in 36 eyes of 18 patients. The therapy group (43 eyes of 22 patients) was administered three daily doses of 100 mg oral NAC. All patients were examined weekly for 1 to 4 months (average, 24 +/- 0.7). A Schirmer-1 test and FBUT were administered at every visit, but mucous fern tests were administered every two weeks. The results of the first and last Schirmer-1 tests, FBUT, and mucous fern test were compared between the therapy and control groups. Student's t and Mann Whitney U tests were used for statistical analysis. RESULTS: FBUT was significantly increased (p < 0.0001), and the mucous fern pattern was also significantly improved (p = 0.0096) in the therapy group. CONCLUSION: NAC is thought to increase FBUT and improve mucous fern pattern by blocking lipid peroxidation in chronic blepharitis. PMID- 11862088 TI - Nonpreserved human amniotic membrane transplantation in acute and chronic chemical eye injuries. AB - PURPOSE: To evaluate the safety and efficacy of nonpreserved amniotic membrane transplantation (AMT) with or without limbal autograft transplantation (LAT) in management of acute and chronic chemical eye injuries. METHODS: Amniotic membrane transplantation or AMT + LAT was performed on nine eyes of seven consecutive patients, five eyes with acute chemical burn and four eyes with limbal stem cell deficiency secondary to previous chemical burn. Nonpreserved amniotic membrane was used in all procedures. RESULTS: Five patients (71.5%) were men and two (28.5%) were women. The average age at the time of surgery was 32.7 +/- 10.9 years (range, 20-45). Mean follow-up after last surgery was 8.9 +/- 3.2 months (range, 6-14). The average epithelial healing time was 24.6 +/- 17.3 days (range, 3-45). At the end of the follow-up period, visual acuity improved in all eyes, inflammation subsided, and the subjective complaints decreased remarkably. CONCLUSION: AMT with nonpreserved amniotic membrane promoted epithelial healing, reduced surface inflammation, increased patient comfort, and decreased the extent and severity of vascularization when used in patients with acute chemical burns. When used in limbal stem cell deficiency owing to past chemical burns, AMT alone or in combination with LAT aided in ocular surface reconstruction. Infectious, inflammatory, or toxic/allergic reactions were not encountered in any patient owing to the use of nonpreserved amniotic membrane. Further studies are required to establish the safety and efficacy of preserved and nonpreserved AMT in ocular surface reconstruction. PMID- 11862090 TI - Limbal stem cell transplantation for the treatment of subepithelial amyloidosis of the cornea (gelatinous drop-like dystrophy). AB - PURPOSE: Subepithelial amyloidosis of the cornea, or gelatinous drop-like corneal dystrophy (GDD), is a severe form of corneal stromal amyloidosis predominantly found in the Japanese population. GDD causes severe visual disturbances with irritating symptoms, and keratoplasty is inevitably complicated by early recurrence. This study was conducted to evaluate the efficacy of limbal stem cell transplantation (LSCT) for the treatment of GDD. METHODS: Nine consecutive eyes of seven patients with GDD were treated with LSCT. Limbal grafts were obtained from cadaver eyes. All eyes underwent lamellar or penetrating keratoplasty simultaneously with LSCT. Intensive epithelial management and immunosuppression therapy was performed. RESULTS: With a mean observation period of 4 years, eight (88.9%) of nine eyes did not show any signs of recurrence. Five grafts remained clear, and seven eyes had an improvement in vision. Glaucoma and rejection were observed in five and three eyes, respectively, and two eyes with glaucoma required surgical intervention. CONCLUSIONS: Limbal stem cell transplantation is effective for the prevention of recurrence in GDD. Control of intraocular pressure and rejection are the keys to long-term maintenance of clear grafts. PMID- 11862089 TI - Age-related prevalence of anterior segment complications in patients with infantile nephropathic cystinosis. AB - PURPOSE: As a result of successful renal transplantation, patients with nephropathic cystinosis are now living into adulthood. As these patients age, anterior segment ocular complications, other than deposition of corneal crystals, become more evident. With our experience with 172 patients followed up at the National Institutes of Health between 1976 and 2000, the prevalence of anterior segment complications in nephropathic cystinosis was determined. METHODS: A cross sectional examination of age-specific prevalence was performed with logistic regression analysis of prevalence change with age. RESULTS: Besides the corneal crystals apparent in all age groups, superficial punctate keratopathy, filamentary keratopathy, severe peripheral corneal neovascularization, band keratopathy, and posterior synechiae with iris thickening and transillumination were noted in the older age groups. The prevalence increased with age for each complication. CONCLUSIONS: As patients with cystinosis grow older, more severe ophthalmic manifestations become evident. It remains to be seen how the prevalence of these complications will be altered by early initiation of oral and topical cysteamine therapy. PMID- 11862091 TI - Objective screening methods for prior refractive surgery in donor tissue. AB - PURPOSE: Laser refractive surgery presents a growing contamination of the available corneal donor pool. There currently is no objective method for screening donor tissue for previous refractive surgery. We evaluated the usefulness of pachymetry and curvature maps in the screening of donor corneas. METHODS: Pachymetry and curvature maps were generated from the Orbscan for 40 normal donor eyes. The average central pachymetry measurement was subtracted from the thinnest average midperipheral pachymetry measurement for each map to generate a normal range of pachymetry measurements. For curvature, the average curvature at the 3-mm zone was subtracted from the average curvature at the 7-mm zone to generate a normal range of corneal curvature. The pachymetry and curvature results from 10 donor eyes that had undergone refractive surgery were then compared with the normal range for each technique. RESULTS: The average difference in pachymetry measurements between the midperipheral and central cornea for normal eyes was 0.040 +/- 0.026 mm. Four of 10 corneas that had undergone refractive surgery were outside two standard deviations of this normal range. The average difference in corneal curvature between the 7-mm zone and the 3-mm zone for the healthy eyes was -0.2 +/- 1.0 diopters. Four (40%) of 10 corneas that had undergone refractive surgery were identified with this method. When combined, the pachymetry and curvature methods detected seven (70%) of 10 corneas that had undergone refractive surgery. CONCLUSION: Regional differences in thickness and curvature in donor eyes may provide methods for screening for refractive surgery for myopia. Refinement in mathematical manipulations may further improve the sensitivity of these techniques. PMID- 11862092 TI - Combined surgery, cryotherapy, and mitomycin-C for recurrent ocular surface squamous neoplasia. AB - PURPOSE: To report the outcome of combined excision, cryotherapy, and antimetabolite treatment of recurrent ocular surface squamous neoplasia. METHODS: The patients with recurrent ocular surface squamous neoplasia were treated by excision of lesion, cryotherapy of limbus, and conjunctival margin followed by 0.02% Mitomycin C application at the time of surgery. Patients have been followed up for at least 1 year. RESULTS: A total of five eyes of five patients with recurrent ocular surface squamous neoplasia were treated by combined excision, cryotherapy, and Mitomycin C. Histopathologic diagnosis included invasive squamous cell carcinoma in four cases and squamous dysplasia in one case. No recurrences have been noted for a follow-up period of more than 1 year now. CONCLUSION: Combining excision with cryotherapy and Mitomycin C application at the time of surgery is a very effective therapy for recurrent ocular surface squamous neoplasia. It is relevant for cases in large, poor countries where patients present late and are less likely to come for follow-up care. PMID- 11862093 TI - A laboratory model for microkeratome-assisted posterior lamellar keratoplasty utilizing a running graft suture and a sutureless hinged flap. AB - PURPOSE: To evaluate changes in anterior corneal curvature and graft stability with different sized donor buttons in a laboratory model of posterior lamellar keratoplasty. METHODS: Thirty-six human eye bank corneas (18 donors and 18 recipients) were mounted on an artificial anterior chamber. A manual microkeratome was used to create a hinged anterior lamellar keratectomy. A 7.0-mm diameter posterior lamellar disk (posterior stroma, Descemet's membrane, and endothelium) was then trephinated from the recipient cornea. Three different sizes (7.0, 7.25, and 7.5 mm) of donor buttons were compared. They were sutured into the recipient bed with a running 10-0 nylon suture and covered by the host corneal flap. The flap was replaced without sutures. The resulting endokeratoplasty was analyzed by computerized videokeratography and tonometry. RESULTS: Regular postoperative astigmatism was present in all cases. There was an average increase in astigmatism of 1.47 +/- 1.49 diopters (D) postoperatively. The mean change in the average keratometry readings was -5.12 +/- 6.12 D. The grafts and flaps maintained watertight seals with average leak pressures of 66.9 +/- 46.4 mm Hg. Although donor buttons oversized by 0.5 mm had the least change in average keratometry reading, those oversized by 0.25 mm had the best stability at high pressure. All groups had little change in astigmatism. CONCLUSION: The optimal sized button of those tested would be either oversized by 0.25 or 0.5 mm. This new surgical technique may result in lower risk of high and irregular astigmatism in the management of corneal endothelial disorders. PMID- 11862094 TI - Separation of corneal stroma and Descemet's membrane during deep lamellar keratoplasty. AB - PURPOSE: To study the interface of the corneal stroma and Descemet's membrane and to report the histologic findings of the deep corneal stromal tissue removed during deep lamellar keratoplasty (DLKP). METHODS: The deep stromal tissues of four corneas were removed during DLKP and were examined histologically by light microscopy (LM) and transmission electron microscopy (TEM). RESULTS: Descemet's membrane appeared as a smooth surface under the surgical microscope during the removal of deep corneal stroma. Two of the four excised tissues showed a thin layer attachment to the deep stromal tissue by LM and TEM. The layer measured approximately 3.0 to 4.0 microm and was observed as a striated structure attached to the stromal tissue of both specimens. The other two specimens contained only stromal tissue. CONCLUSIONS: Separation of the deep stromal tissue from Descemet's membrane may occur within the Descemet's membrane, and the separation is probably between the anterior banded and the posterior nonbanded layer of Descemet's membrane in some cases during DLKP. The lamellar structure of the delaminated region suggests a mechanically weak segment of Descemet's membrane. The smooth surface of Descemet's membrane observed under a surgical microscope is not the actual interface of corneal stroma and Descemet's membrane. PMID- 11862095 TI - Intraocular dexamethasone delivery system for corneal transplantation in an animal model. AB - PURPOSE: To assess the efficacy of a new intraocular biodegradable polymer dexamethasone drug delivery system (DEX DDS) in a high-risk corneal transplantation model. METHODS: Lewis rats that received orthotopic corneal transplants (Balb/c mice donors) were divided into three groups (six rats in each); group 1 received no treatment and served as controls, group 2 was treated with 0.1% betamethasone eyedrops three times daily for 6 weeks, and group 3 received DEX DDS in the anterior chamber at the time of transplantation. RESULTS: All grafts in the untreated control group were rejected within 8 days. In the betamethasone eyedrop group, five eyes (83%) were rejected during the 8-week study period. None of the grafts in the DEX DDS group was rejected. The administration of DEX DDS significantly prolonged the survival rate of the corneal grafts (p < 0.001, log-rank test). CONCLUSION: DEX DDS is effective in suppressing graft rejection in high-risk corneal transplantation. PMID- 11862096 TI - Development and application of an in vitro susceptibility test for Acanthamoeba species isolated from keratitis to polyhexamethylene biguanide and chlorhexidine. AB - PURPOSE: To develop a reliable in vitro drug susceptibility test against Acanthamoeba isolates and to determine the minimum cysticidal concentration (MCC) of the drug. METHODS: Doubling dilutions of polyhexamethylene biguanide (PHMB) from 3,200 microg/mL to 3.125 microg/mL and chlorhexidine from 3,200 microg/mL to 1.5625 microg/mL were made in Durham tubes and tested against cysts of 19 Acanthamoeba isolates from keratitis. After the exposure to the drugs for 48 hours, the cysts were washed free of drugs by centrifugation. The deposit (cysts) was cultured on nonnutrient agar plates seeded with heat-killed Escherichia coli. The growth of trophozoites from cysts exposed to each of the dilution was recorded by microscopy to estimate the MCC of the drug. RESULTS: The minimum cysticidal concentration of PHMB varied from 25 microg/mL to 100 microg/mL and that of chlorhexidine varied from 1.56 microg/mL to 100 microg/mL. The mean MCC value for PHMB was 55.26 microg/mL and that for chlorhexidine 32.81 microg/mL. Minimum cysticidal concentration 50 (MCC50) of PHMB and chlorhexidine on Acanthamoeba isolates was 50.0 microg/mL and 25.0 microg/mL, respectively. Anti Acanthamoeba activity of chlorhexidine was higher than that of PHMB and this was statistically significant (p = 0.036). The end point of the results of this method was the detection of the viable cysts undergoing excystment and multiplication of trophozoites with a reproducible and clear-cut estimation of the MCC of PHMB and chlorhexidine. CONCLUSION: The in vitro method described can be used as a standard test for assay of MCC of drugs on Acanthamoeba isolates and to study the susceptibility pattern of newer water-soluble anti-Acanthamoeba drugs. PMID- 11862097 TI - Evidence of apoptotic cell death in keratoconus. AB - PURPOSE: To determine the potential role of apoptosis in the noninflammatory degeneration characteristic of keratoconus. METHODS: Four normal corneas and 16 keratoconus corneas were obtained as archival specimens. Tissues were examined histopathologically for TUNEL immunoreactivity to detect the presence of DNA fragmentation. Tissues were also subjected to single-stranded DNA (ssDNA) analysis, a more apoptosis-specific stain. RESULTS: Normal corneas exhibited fewer than five TUNEL-positive epithelial cells per section, these being very lightly stained. All 16 keratoconus corneas demonstrated extensive, intense TUNEL staining in at least one layer. Fifteen of 16 exhibited staining in the epithelial layer, 11 of 16 in the stromal layer, and 13 of 16 in the endothelial layer, whereas 10 of 16 keratoconus cases demonstrated TUNEL immunoreactivity in all three layers. The ssDNA stain was also positive and evident in all three layers of the cornea, although to a lesser degree than the TUNEL assay. CONCLUSIONS: The noninflammatory nature of keratoconus, coupled with the TUNEL in situ results, suggests apoptosis as a mode of cell death in this degenerative disease. PMID- 11862098 TI - Expression of IL-10 and TNF-inhibitor genes in lacrimal gland epithelial cells suppresses their ability to activate lymphocytes. AB - PURPOSE: To determine whether the expression of either interleukin-10 (IL-10) or tumor necrosis factor (TNF) inhibitor genes in transduced rabbit lacrimal gland epithelial cells suppresses lymphocyte proliferation in an autologous mixed cell reaction, an apparent in vitro model of autoimmune dacryoadenitis. METHODS: Purified lacrimal gland epithelial cells, transduced with an adenovirus vector carrying either viral IL-10 or TNF-inhibitor genes, were used to study their effects on the proliferation of autologous lymphocytes as monitored by 3H thymidine incorporation in a mixed cell reaction. After transduction, both epithelial cells and lymphocytes were cultured separately for 2 days and then epithelial cells were irradiated. Equal numbers of both cell types were then cocultured together for 5 days. Cocultures were pulsed with 3H-thymidine and isotope incorporation was determined. Gene expression was detected by enzyme linked immunosorbent assay and Western blots. RESULTS: Lymphocyte proliferation was stimulated by epithelial cells and 3H-thymidine incorporation was significantly greater in these cocultures than in controls. The proliferation was significantly diminished in the presence of transduced cells producing either IL 10 or TNF inhibitor. CONCLUSIONS: Transduction of lacrimal gland epithelial cells with adenovirus vectors encoding for either IL-10 or TNF-inhibitor proteins leads to expression of functional proteins capable of suppressing lymphocyte proliferation. Thus, lacrimal gland epithelial cells are a plausible target for gene therapy methods meant to produce immunoregulatory proteins. PMID- 11862099 TI - Changes of nuclear matrix in long-term culture of limbal epithelial cells. AB - PURPOSE: This study was undertaken to investigate the changes of nuclear matrix in long-term culture of rabbit limbal epithelial cells. METHODS: Epithelial cells outgrown from limbal basal epithelium were serially cultivated. Nuclear matrices of early and late passages were extracted for morphologic study and protein analysis by two-dimensional gel electrophoresis and immunoblotting. RESULTS: Differential growth and changes in morphology were observed in limbal epithelial cells of early and late passages. Cytokeratin type 3 was expressed in cells of later passages, indicating corneal cell differentiation during the long-term culture. These cells also showed reduced density of nuclear matrix fibrils and thinning of nuclear lamina. They were shown by two-dimensional gel electrophoresis to have lost most nuclear matrix proteins, including lamin A/C and proliferating cell nuclear antigen. However, five new protein entities were also expressed. CONCLUSION: The nuclear matrix appeared to change along with limbal epithelial cell differentiation in culture. Whether such changes may affect the growth and viability of limbal cells after transplantation requires in vivo tissue analysis. PMID- 11862100 TI - Posterior amorphous corneal dystrophy: ultrasound biomicroscopy findings in two cases. AB - We present two cases of posterior amorphous corneal dystrophy in members of the same family. We correlated the clinical findings with refraction, topography, and ultrasound biomicroscopy (UBM). This is the first report of UBM findings in amorphous corneal dystrophy and we describe the depth of stromal opacification measured in this exam. Additional cases will be of great help to reinforce these findings. PMID- 11862101 TI - Exacerbation of Avellino corneal dystrophy after laser in situ keratomileusis. AB - PURPOSE: To report a case of Avellino corneal dystrophy that increased in severity 1 year after uncomplicated laser in situ keratomileusis (LASIK) for myopia. METHODS: Avellino dystrophy was confirmed by polymerase chain reaction sequencing of DNA from the patient and her parents. RESULTS: Best spectacle corrected visual acuity decreased from 20/20 to 20/30 12 to 20 months after LASIK owing to opacities that appeared centrally in the corneal stroma and the LASIK flap and remaining posterior stroma interface. CONCLUSIONS: LASIK is contraindicated in patients with Avellino corneal dystrophy because vision may be reduced by corneal opacities that appear in the interface of the flap and remaining posterior stroma postoperatively. PMID- 11862102 TI - Acute scleral thinning after pterygium excision with intraoperative mitomycin C: a case report of scleral dellen after bare sclera technique and review of the literature. AB - PURPOSE: To describe a patient with scleral dellen after pterygium excision with intraoperative mitomycin C. METHODS: Case report and MEDLINE review of the medical literature on scleral dellen after bare sclera technique. RESULTS: A 48 year-old man had a left nasal pterygium excised by the bare sclera technique with intraoperative mitomycin C. Eight days after surgery, the patient noticed a small black spot in the bare sclera area with mild irritation. Slit-lamp examination revealed a focal area of extreme thinning, centered on the nonepithelialized bare sclera, surrounded by edematous conjunctiva. The ciliary body was visible through the thin and dry scleral lesion. After topical lubricant therapy, the scleral lesion appeared normal thickness and white in color 3 days later. Therapy was continued until the sclera epithelialized. CONCLUSIONS: Scleral dellen is an early postoperative complication of bare sclera technique owing to delayed conjunctival wound closure. Hydration of the thinned sclera will rapidly thicken it. However, medical therapy should be continued until the surrounding conjunctiva has flattened and the sclera has epithelialized. Surgical wound closure is an alternative management and may be the way to prevent scleral dellen formation after bare sclera technique. All patients after bare sclera surgery should be followed up until the conjunctival wound has healed. If delayed healing is found, frequent artificial tears, patching, or surgical intervention is necessary. PMID- 11862104 TI - Postkeratoplasty endophthalmitis by Alcaligenes faecalis: a case report. AB - PURPOSE: To describe a postkeratoplasty endophthalmitis owing to Alcaligenes faecalis that resolved with medical management. METHODS: Case report and review of literature. RESULTS: To best of our knowledge, postkeratoplasty endophthalmitis caused by A. faecalis has not been reported in literature. The source of infection was found to be the donor tissue. The organism isolated was resistant to gentamicin, which was the only antibiotic present in the storage medium (McCarey and Kaufman medium). The patient was treated aggressively with topical, systemic and intravitreal antibiotics. The endophthalmitis resolved completely within two weeks. CONCLUSION: A. faecalis is a gram negative, non spore forming rod that can cause endophthalmitis if donor tissue is contaminated with the organism. Clinical suspicion, review of bacterial culture and sensitivity report from donor tissue, anterior chamber and vitreous taps, along with early therapy with appropriate antibiotics can lead to resolution of infection. PMID- 11862103 TI - More than tears in your eyes (Exophiala jeanselmei keratitis). AB - PURPOSE: To describe a patient with Exophiala jeanselmei keratitis. METHODS. CASE REPORT: One patient with persistent corneal infiltrate that developed several days after a minor ocular trauma from an onion slice. RESULTS: Culture plates from corneal scraping showed a growth of the yeast Exophiala jeanselmei, a rare causative agent of ocular infection. CONCLUSIONS: Whenever a corneal abscess does not improve with the usual antibiotic treatment, a thorough ophthalmic history should be taken to determine whether there was a recent ocular trauma. If the trauma was caused by a plant material, the physician should raise the possibility of an unusual fungal infection. PMID- 11862105 TI - Corneal laceration and intraocular foreign body in a post-LASIK eye. AB - PURPOSE: To describe a case of full-thickness corneal laceration and intraocular foreign body in an eye that underwent laser-assisted in situ keratomileusis (LASIK). METHODS: A 43-year-old man underwent uneventful bilateral LASIK. Seventeen months after LASIK, the patient suffered penetrating trauma to the right eye while hammering metal without safety glasses. Examination on the night of the injury showed an uncorrected visual acuity of 20/30 in the right eye. Slit lamp biomicroscopy showed a 2.4-mm full-thickness peripheral corneal laceration at the 11-o'clock position extending over the edge of the previous LASIK flap and a foreign body partially embedded in the superior iris. He was started on prophylactic topical and intravenous antibiotics. Under general anesthesia, the corneal laceration was repaired, and the intraocular foreign body was removed without any complications. RESULTS: At the follow-up visit 9 months after the surgery, the uncorrected visual acuity was 20/25 in the right eye. On slit-lamp examination, the corneal sutures and the well-positioned LASIK flap were in place. CONCLUSION: Penetrating trauma of the cornea did not lead to flap-related complications in this post-LASIK eye. Repair of the corneal laceration and removal of the intraocular foreign body after LASIK were managed without complications. Care must be taken to try not to manipulate or lift the LASIK flap during corneal laceration repair. PMID- 11862106 TI - Progressive ulcerative keratitis related to the use of topical chlorhexidine gluconate (0.02%). AB - PURPOSE: To report a case of progressive ulcerative keratitis related to the use of topical chlorhexidine gluconate 0.02%. METHODS: A 45-year-old woman was treated for presumed Acanthamoeba keratitis with chlorhexidine gluconate 0.02% and propamidine 0.1% eyedrops. RESULTS: After using the chlorhexidine and propamidine eyedrops for 8 weeks, the patient developed a near total loss of the corneal epithelium and a progressive ulcerative keratitis, which eventually required penetrating keratoplasty. Histopathologic examination of the corneal button showed ulceration and loss of Bowman's membrane, massive loss of keratocytes with apparent apoptosis, and loss of the endothelial cells with inflammatory cells adherent to the remaining cells. These histopathologic findings are similar to those seen in Hibiclens keratopathy. Hibiclens contains chlorhexidine 4% with a detergent. No organisms were seen in sections stained with modified Ziehl-Neelsen, gram, Grocott, or periodic acid-Schiff, and immunohistochemistry showed no significant findings. CONCLUSION: We think that the topical use of chlorhexidine gluconate 0.02% may have resulted in a progressive ulcerative keratitis in our patient. PMID- 11862107 TI - Keratitis caused by Verticillium species. AB - PURPOSE: To report a case of fungal keratitis caused by Verticillium species. METHODS: A 50-year-old man developed pain, redness, and an infiltrate in his left eye and had no history of trauma. The cornea showed superficial, white, stromal infiltrates and epithelial ulceration with a dendritic margin. The clinical features suggested herpetic keratitis, and the patient was treated with topical antiviral medication. Two weeks later, his condition deteriorated. Examination of the left eye showed stromal infiltrates with a feathery margin and epithelial ulceration with its covering white exudates. Corneal scrapings were taken for direct microscopic examination and culture. RESULTS: Corneal scraping showed the presence of fungal filaments. The fungus was identified as Verticillium species. Topical amphotericin B and systemic fluconazole were started after discontinuing the antiviral treatment. Clinically, the inflammation subsided during the 3 weeks after treatment. CONCLUSION: This is a rare case of infectious keratitis caused by Verticillium species. Rare species of fungal infection should be considered in the differential diagnosis of stromal keratitis refractive to conventional medical treatment. PMID- 11862109 TI - Rho-kinase as a novel therapeutic target in treatment of cardiovascular diseases. AB - Rho-kinase has been identified as one of the effectors of the small GTP-binding protein Rho. Accumulating evidence has demonstrated that the Rho/Rho-kinase mediated pathway plays an important role in various cellular functions, not only in vascular smooth muscle contraction but also in actin cytoskeleton organization, cell adhesion and motility, cytokinesis, and gene expressions, all of which may be involved in the pathogenesis of arteriosclerosis/atherosclerosis. Indeed, animal experiments have demonstrated that Rho-kinase inhibitors effectively suppress coronary artery spasm and that long-term inhibition of Rho kinase inhibits the development of coronary arteriosclerotic lesions and even causes regression of coronary vascular lesions in vivo. Recent clinical studies also have demonstrated the inhibitory effect of a Rho-kinase inhibitor on coronary artery spasm in patients with vasospastic angina and on exercise-induced myocardial ischemia in patients with stable effort angina with adequate safety. It is possible that Rho-kinase is also involved in the pathogenesis of other forms of cardiovascular diseases. Thus, Rho-kinase could be regarded as a novel therapeutic target in treatment of cardiovascular diseases. PMID- 11862110 TI - Is the apico-basal gradient larger than the transmural gradient? PMID- 11862111 TI - Modulation of matrix metalloproteinase-1, its tissue inhibitor, and nuclear factor-kappa B by losartan in hypercholesterolemic rabbits. AB - Upregulation of angiotensin II receptor, may be involved in the initiation and progression of atherosclerosis. To examine the contribution of AT1 receptor in the expression of matrix metalloproteinase-1 (MMP-1) and its tissue inhibitor (TIMP-2) in lipid-deposited arterial tissues, New Zealand white rabbits were given high-cholesterol chow (with losartan 25 mg/d or vehicle) for 10 weeks. Losartan reduced the areas of sudanophilia in the aorta of rabbits fed high cholesterol diet (p < 0.01 vs. control). Losartan also significantly decreased the enhanced mRNA expression of MMP-1 and TIMP-2 in aortas of rabbits with high cholesterol diet. Losartan-treated rabbits revealed a reduction in immunohistochemical expression of MMP-1, whereas TIMP-2 expression became localized to the intima. In addition, losartan treatment reduced the activation of NF-kappa B by inhibiting the degradation of its inhibitor I kappa-B alpha. These observations demonstrate that AT1 receptor blockade with losartan reduces lipid deposition and exerts potent inhibitory effects on NF-kappa B activation and modulates the expression of MMP-1 and TIMP-2 in hypercholesterolemic rabbits. PMID- 11862112 TI - Effects of cromakalim and glibenclamide on arteriolar and venular diameters and macromolecular leakage in the microcirculation during ischemia/reperfusion. AB - We studied changes in arteriolar and venular diameters and macromolecular leakage altered by ischemia/reperfusion (I/R) and topically applied histamine after I/R and how these changes were modulated by cromakalim (KATP-channel opener) and glibenclamide (KATP-channel blocker). Golden hamsters were prepared for intravital microscopy of the cheek pouch. Ischemia was induced by an inflatable silicon rubber cuff mounted around the neck of the cheek pouch prepared for intravital microscopy. Saline, histamine, cromakalim, and glibenclamide were applied in the superfusion solution. FITC-dextran was injected i.v. 30 min before initiation of ischemia as a marker of macromolecular leakage. Cromakalim 10(-6) M, but not 10(-8) M, caused arteriolar dilation in ischemic and normal (nonischemic) preparations, and glibenclamide, 10 -10) M and 10(-8) M, had no effects on vessel diameters. Application of cromakalim 10(-6) M increased arteriolar diameter (+54%) and macromolecular leakage in normal and nonischemic cheek pouches and had an additive effect on macromolecular leakage in ischemic (I/R) preparations but had no effect on histamine-induced increase in macromolecular leakage. Glibenclamide, 10(-10) M and 10(-8) M, inhibited I/R induced but not histamine-induced increases in macromolecular leakage. We concluded that cromakalim may increase macromolecular leakage. This effect is additive to I/R-induced leakage suggesting that stimulation of KATP-channels could take part in the regulation of macromolecular leakage in postcapillary venules. The KATP-blocker glibenclamide inhibited I/R-induced but not histamine induced macromolecular leakage at concentrations that had no constricting effect on arterioles, and therefore, it cannot be excluded that glibenclamide reduced plasma leakage by some unknown mechanism. PMID- 11862113 TI - 17-beta estradiol preserves endothelial cell viability in an in vitro model of homocysteine-induced oxidative stress. AB - High levels of homocysteine (Hcy) accelerate endothelial cell damage by producing hydrogen peroxide (H(2)O(2)). We investigated whether 17-beta estradiol may prevent the accelerated rate of endothelial cell detachment and reduced cell viability in cultured endothelial cells challenged with Hcy. Cultured bovine aortic endothelial cells (BAEC) were incubated for 72-h with either vehicle (alcohol) or different concentrations of 17-beta estradiol (1 nM [1E2] and 10 nM [10E2]) before being challenged with 0.5 mM of Hcy. Cell viability and H(2)O(2) levels were evaluated at 30 min, 1-, 2-, 4-, 8-, and 24-h after adding Hcy. Cell suspensions were frozen in liquid nitrogen and used later for spectrophotometric measurement of intracellular glutathione (GSH) levels. Cell viability 24 h after Hcy administration was significantly lower in vehicle versus 1 nM and 10 nM 17 beta estradiol (44 +/- 5% vs. 70.66 +/- 4%, [p < 0.001] and 79 +/- 5% respectively, [p < 0.001]). H(2)O(2) levels were higher in vehicle (1 +/- 0.05 microM) compared with 1E2 and 10E2 (0.8 +/- 0.1 microM, p = 0.02 and 0.1 +/- 0.05 microM, respectively, p < 0.001), whereas GSH content was increased in 1E1 and 10E2 versus control (27.69 +/- 4.6 nM/10(6) cells and 43.49 +/- 5.5 nM/10(6) cells vs. 13.33 +/- 1.5 nM/10(6) cells, p < 0.001). Bovine aortic endothelial cells treatment with 17-beta estradiol (0, 1, and 10 nM) and 0.1 mmol buthionine sulfoximine, an inhibitor of gamma-glutamylcysteine synthase, abolished the beneficial effects of estradiol alone on cell viability, GSH content, and H2O2 generation. Estradiol prevents Hcy-induced endothelial cell injury by increasing the intracellular content of GSH. PMID- 11862114 TI - Effect of nitric oxide synthase inhibitor N(omega) nitro-L-arginine methyl ester on relaxant responses to calcium channel antagonists in isolated aortic rings from Dahl normotensive and hypertensive rats. AB - Differences exist between the pharmacological actions of calcium channel antagonists in blood vessels from hypertensive versus normotensive animals. In this investigation, we have examined the impact of nitric oxide synthase inhibitor N(omega) nitro-L-arginine methyl ester (L-NAME) on relaxant responses produced by the calcium channel antagonists (nifedipine, diltiazem, and mibefradil) in isolated aortic rings from Dahl salt-resistant normotensive (SRN) and salt-sensitive hypertensive (SSH) rats on a 4% salt diet. Morphological examination of the aortic rings revealed significantly larger lumen area, smooth muscle wall thickness, and perimeter in vessels of SSH rats versus SRN rats. Rank order potency for the antagonists was nifedipine > mibefradil > or = diltiazem in aortic rings from SRN rats, but mibefradil was found to be the most efficacious. The rank order potency for the antagonists in aortic rings from SSH rats was nifedipine > diltiazem > mibefradil, although all three drugs showed similar efficacy. The presence of L-NAME attenuated relaxations elicited by the antagonists in aortic rings from SRN rats. Treatment of tissues with L-NAME significantly reduced maximal response and decreased pIC(50). The presence of L NAME had no effects on concentration-response curves to nifedipine and diltiazem in aortic rings from SSH rats, but it significantly attenuated relaxant responses of mibefradil. Our current results support the view that these calcium channel antagonists produce relaxations by mechanisms that are sensitive and insensitive to L-NAME. Moreover, the component insensitive to L-NAME was lacking in tissues from SSH rats for nifedipine and diltiazem but not mibefradil. PMID- 11862115 TI - Tetrahydrobiopterin improves impaired endothelium-dependent forearm vasodilation in patients with heart failure. AB - Endothelium-dependent vasodilation is impaired in patients with chronic heart failure (CHF). However, the mechanisms responsible for this effect are not fully understood. The vasodilator response to acetylcholine (ACh) has been used to examine the endothelium-dependent vasodilation in humans and is known to be mediated by nitric oxide (NO). The impaired production of NO or an increase in its degradation is thought to be responsible for the endothelial dysfunction in CHF. The aim of this study was to determine whether the decrease in availability of tetrahydrobiopterin (BH(4)), an essential cofactor of NO synthase, contributes to the impairment of endothelium-dependent vasodilation in patients with CHF. Fourteen patients with CHF (New York Heart Association functional class II-IV, age: 59 +/- 4 years, ejection fraction: 28 +/- 3%) and seven age-matched control subjects were examined. Forearm blood flow (FBF) was measured by plethysmography during an intra-arterial infusion of a graded dose of ACh (4, 8, and 16 microg/min) and sodium nitroprusside (SNP) (0.8, 1.6, and 3.2 microg/min). These procedures were repeated during a co-infusion of BH(4) (400 microg/min). The forearm vasodilator response to ACh was significantly enhanced during co-infusion of BH(4) in patients with CHF, whereas no effect was observed in healthy subjects. In contrast, the response to SNP was not affected by BH(4) in either group. The administration of BH(4) did not alter the baseline FBF in either group. These results suggest that an acute administration of BH(4 ) improves endothelium-dependent forearm vasodilation in patients with CHF. PMID- 11862116 TI - Severe nutritional restriction in pregnant rats aggravates hypertension, altered vascular reactivity, and renal development in spontaneously hypertensive rats offspring. AB - Epidemiologic studies suggest that intrauterine undernutrition plays an important role in the development of arterial hypertension in adulthood. The aim of the current study was to evaluate whether severe nutritional restriction during pregnancy can aggravate hypertension, vascular reactivity changes, and renal development in spontaneously hypertensive rat (SHR) offspring. To investigate the potential existence of gender differences, both male and female offspring of pregnant SHRs on a restricted diet were studied in adulthood. Female pregnant SHRs were fed either normal or 50% of the normal intake diets, during the whole gestational period. Arterial blood pressure and nephron number were determined. Norepinephrine, acetylcholine, and sodium nitroprusside responses in isolated aortic rings from the offspring (male and female, when they reached adulthood) were also evaluated. In the SHR offspring (male and female) the intrauterine undernutrition further increased the blood pressure levels, increased the response to norepinephrine, and decreased the response to acetylcholine, without altering the response to sodium nitroprusside. In addition, it induced a decrease in the number of nephrons in the kidney from adult offspring. In conclusion, fetal undernutrition aggravates hypertension and the endothelial dysfunction along with an impairment of renal development in both male and female SHRs. PMID- 11862117 TI - Alterations of the nitric oxide pathway in cerebral arteries from spontaneously hypertensive rats. AB - Hypertension-associated alterations of the nitric oxide (NO) pathway were analyzed in middle cerebral arteries (MCA) from normotensive (WKY) and hypertensive (SHR) rats. The vasoconstrictor response to prostaglandin F2alpha (PGF(2 alpha), 30 and 100 microM) was smaller in MCA from SHR than from WKY. Endothelium-dependent relaxations to bradykinin (1 nM-10 microM) or acetylcholine (10 microM) were similar in MCA from both strains, whereas the endothelium independent response to sodium nitroprusside (1 nM-0.1 mM) was smaller in MCA from SHR. L-arginine (L-Arg, 10 microM) similarly inhibited the vasoconstrictor responses in both strains; however, the inhibitory effect of 100 microM of L-Arg was greater in MCA from SHR. N(omega)-nitro-L-arginine methyl ester (L-NAME, 100 microM), but not aminoguanidine (100 microM) or 7-nitroindazole (10 microM), increased basal tone, potentiated the PGF(2 alpha)-induced vasoconstrictor responses and reduced the bradykinin-elicited relaxation in a similar way in MCA from WKY and SHR. N(omega)-nitro-L-arginine methyl ester also antagonized the inhibitory effect of 10 microM of L-Arg. Incubation for 5 h with lipopolysaccharide (10 microg/ml) similarly reduced the response to PGF(2 alpha) in MCA from WKY and SHR; this reduction was antagonized by dexamethasone (1 microM). Cerebral arteries expressed endothelial (eNOS) and neuronal (nNOS) NO synthase similarly in both strains, but inducible NOS (iNOS) expression was more evident in SHR. Lipopolysaccharide increased iNOS expression in both strains to a similar level. The basal constitutive NOS (cNOS) and iNOS activities were similar in arteries from WKY and SHR. Lipopolysaccharide increased iNOS activity only in arteries from SHR. These results indicate that hypertension did not impair endothelial NO production by NOS activation but induced an up-regulation of basal iNOS expression. PMID- 11862118 TI - Early benefits of pravastatin to experimentally induced atherosclerosis. AB - There is little information regarding the time of hypolipidemic treatment of changes in atherosclerotic plaque, tissue cholesterol content, and also for the recovery of endothelial function. To assess the early effects of lipid-lowering treatment on these parameters, six groups of New Zealand male rabbits were studied. Animals in groups I and II were fed regular chow; groups III and IV received a 12-week 0.5% cholesterol diet followed by 12 weeks of 0.05% cholesterol diet. Finally, groups V and VI were fed a 12-week 0.5% cholesterol diet and were then shifted to a regular diet for 12 weeks. During the last four weeks, the rabbits in groups I, III, and V received low-dose pravastatin (2 mg/day), added to the diet. Group IV animals had the highest cholesterol plasma levels (vs. groups I, II, III, and V, p < 0.01) and presented atherosclerotic plaques in a more advanced stage. Nonatherogenic diet was insufficient to restore endothelial function in animals previously fed cholesterol-enriched diets (groups IV and VI). Conversely, pravastatin treatment promoted significant improvement in endothelial function and reduced the progression of atherosclerosis. Marked increase in cholesterol content was seen in aorta and liver in response to the atherogenic diet. However, neither treatment with pravastatin nor nonatherogenic diet was capable of modifying the tissue cholesterol content. Our study supports the hypothesis that the early use of statins can attenuate the progression of atherosclerosis and ameliorate endothelial function. In addition, significant changes in the tissue cholesterol pool probably need a longer period of treatment. PMID- 11862119 TI - Response of rat thoracic aorta to F(2)-isoprostane metabolites. AB - SUMMARY: This study was undertaken to investigate the vascular actions (contraction and relaxation) of the F(2)-isoprostane metabolites 15-keto-15-F(2t) IsoP, 2,3-dinor-15-F(2t)-IsoP, and 2,3-dinor-5,6-dihydro -15-F(2t)-IsoP in comparison with 15-F(2t)-IsoP on the rat thoracic aorta. 15-keto-15-F(2t)-IsoP induced a vasoconstriction in a concentration-dependent manner with a pD(2) value of 5.80 +/- 0.05, whereas 2,3-dinor-15-F(2t)-IsoP and 2,3-dinor-5,6-dihydro-15 F(2t)-IsoP had no effect. The parent compound 15-F(2t)-IsoP was more potent (pD(2) value: 6.46 +/- 0.1). Endothelium removal had no influence on the contraction to 15-keto-15-F(2t)-IsoP. GR32191 (a TP-receptor antagonist) concentration-dependently inhibited the contraction induced by 15-keto-15-F(2t) IsoP, with a significant decrease in the E(max) values for GR32191 10(-7) M. Pretreatment with 2,3-dinor-15-F(2t)-IsoP and 2,3-dinor-5,6-dihydro-15-F(2t)-IsoP induced no alteration of 15-F(2t)-IsoP concentration-response curves. In contrast, 15-keto-15-F(2t)-IsoP pretreatment competitively inhibited the response to 15-F(2t)-IsoP. When concentration ratios of EC(50) values were used, a Schild regression of this data was linear with a slope of 0.974 and a pA(2) value of 6.13. 15-keto-15-F(2t)-IsoP at high concentrations caused a weak concentration dependent relaxation of rat aorta rings contracted with U46619 (3.10(-8) M) that was not modified in the absence of endothelium. In contrast, 2,3-dinor-15-F(2t) IsoP and 2,3-dinor-5,6-dihydro-15-F(2t)-IsoP induced no vasodilation. In conclusion, among the F(2)-isoprostane metabolites, 2,3-dinor-15-F(2t)-IsoP and 2,3-dinor-5,6-dihydro-15-F(2t)-IsoP did not cause vasorelaxation or vasoconstriction on the rat thoracic aorta. In contrast, 15-keto-15-F(2t)-IsoP mediates contraction through activation of TP-receptors, probably as a partial agonist, and induces a weak endothelium-independent relaxation at high concentrations. PMID- 11862120 TI - Sex does not seem to influence the formation of aortic lesions in the P-407 induced mouse model of hyperlipidemia and atherosclerosis. AB - Coronary heart disease secondary to atherosclerosis is the leading cause of death for men in the United States. Using a new, nontransgenic, non-fat-fed mouse model of hyperlipidemia and atherosclerosis developed in our laboratory, we investigated the effect of sex on lipid profiles and subsequent aortic atherosclerotic lesion formation. Female and male C57BL/6 mice, which consumed a low-fat diet, were treated with either normal saline or poloxamer 407 (P-407), a triblock copolymer comprised of poly(oxyethylene) and poly(oxypropylene) units, for 4 months. Blood samples were obtained at 0, 1, 2, 3, and 4 months, whereas hearts and livers were harvested only at 4 months, because this model requires approximately 4 months for significant atheroma formation. P-407-treated mice of either sex demonstrated a profound increase in plasma cholesterol and triglyceride; at 3 and 4 months the plasma lipids were significantly (p < 0.05) higher for male mice compared with female mice. Aortas retrieved from P-407 treated mice of either sex after 4 months demonstrated a significant (p < 0.001) increase in the mean atherosclerotic lesion size compared with their respective saline-treated controls, but there was no significant (p > 0.05) difference between lesion sizes for P-407-treated male mice (1.02 +/- 0.074 x 10(5) microm(2)) compared with P-407-treated female mice (1.14 +/- 0.28 x 10(5) microm(2)). Livers harvested at 4 months from either sex of P-407-treated mice displayed no damage to hepatocytes but increased proliferation of macrophages (Kupffer cells), which contained sequestered lipids. Thus, male C57BL/6 mice form atherosclerotic lesions as extensive as female mice in the P-407 mouse model of atherosclerosis. PMID- 11862121 TI - Limited antifibrillatory effectiveness of clinically relevant concentrations of class I antiarrhythmics in isolated perfused rat hearts. AB - The Langendorff-perfused rat heart with regional ischemia is increasingly used for evaluating drugs for prevention of phase-1, ischemia-induced ventricular fibrillation (VF). Surprisingly, the effectiveness of Class I antiarrhythmics has not been characterized in this model. One lower and one higher concentration of quinidine (0.79 and 7.90 microM), lidocaine (3.88 and 12.93 microM), and flecainide (0.74 and 1.48 microM), representing the peak unbound plasma and total blood concentrations, respectively, at "therapeutic" dosage, were evaluated. The left main coronary artery was occluded for 30 min to elicit phase-1 VF, after which reperfusion-induced VF was examined. In hearts perfused with Krebs' solution containing 3 mM K(+), the higher concentrations of quinidine and lidocaine reduced the incidence of phase-1 VF from 92% to 0% and 17% respectively, (each p < 0.05). The lower drug concentrations were ineffective. Flecainide was equi-effective at low and high concentrations, with VF incidence reduced from 92% to 17% (p < 0.05). Neither low nor high concentrations of any of the drugs affected the incidence of reperfusion-induced VF. Using hearts perfused with Krebs' containing 5 mM K(+), sufficient to substantially reduce control phase-1 VF incidence, the experiment was repeated to test for possible proarrhythmic activity. None of the three drugs increased arrhythmia incidence. In this model, it was not possible to suppress ischemia-induced and reperfusion induced VF with flecainide, lidocaine, or quinidine at concentrations equivalent to peak unbound plasma levels after clinical administration. This may explain the lack of clinical benefit with these drugs against sudden cardiac death. Because none of the drugs were proarrhythmic in ischemic hearts in which arrhythmia susceptibility had been lowered by high K(+), it would seem that clinical proarrhythmia seen with these drugs may not be related to exacerbation of phase 1, ischemia-induced VF. PMID- 11862122 TI - Does recombinant human interleukin-11 exert direct electrophysiologic effects on single human atrial myocytes? AB - Recombinant human interleukin-11 (rhIL-11) treatment given to alleviate side effects of cancer therapy is associated with an increased susceptibility to atrial arrhythmias in elderly patients. To elucidate the mechanism underlying this action, we investigated the direct electrophysiologic effect of rhIL-11 on single human atrial myocytes (HuAM) using the patch-clamp technique. Action potentials (AP) at different driving rates were recorded in the perforated-patch configuration, and L-type calcium current (I(Ca,L)), outward potassium currents (I(to) and I(K)), and the hyperpolarization-activated pacemaker current If were measured in the disrupted whole-cell configuration. At therapeutic concentrations (i.e., 10-100 ng/ml), rhIL-11 did not modify AP parameters and cycle-length dependence of AP duration. I(Ca,L) (measured at 0 mV) was 370 +/- 45 pA in control and 379 +/- 48 pA and 368 +/- 42 pA in the presence of 10 and 50 ng/ml rhIL-11, respectively (p = NS). The amplitude and activation of I(to) were not modified by rhIL-11 (i.e., I(to) was at +60 mV: 2.1 +/- 0.2 nA in control vs. 1.9 +/- 0.2 nA and 2.1 +/- 0.2 nA in the presence of 10 and 50 ng/ml rhIL-11, respectively, p = NS). Similarly, late currents measured at the end of the pulse were unchanged in the presence of 10 or 50 ng/ml of rhIL-11. If activation was not modified by rhIL-11: maximal current was 173 +/- 34 pA in control and 159 +/- 35 pA and 117 +/- 14 pA in the presence of 10 and 50 ng/ml of rhIL-11, respectively; midpoint activation was -99 +/- 3 mV in control and -98 +/- 4 mV and -94 +/- 2 mV in the presence of 10 and 50 ng/ml of rhIL-11, respectively (p = NS). Thus, it is unlikely that direct alterations of membrane potential and currents of HuAM caused by rhIL-11 are the basis for the genesis of atrial arrhythmias observed in patients treated with this agent. PMID- 11862123 TI - Age-dependent atrial remodeling induced by recombinant human interleukin-11: implications for atrial flutter/fibrillation. AB - Recombinant human interleukin-11 (rhIL-11) has been used to alleviate side effects caused by chemotherapy in patients with neoplastic diseases and as an experimental anti-inflammatory agent. Its use in elderly patients has been associated with increased frequency of atrial flutter/fibrillation (AF). The mechanism of the latter phenomenon was studied in a rat model in vivo. Adult and old Fischer 344 rats fed with either normal diet or sodium-restricted diets were treated with rhIL-11 or vehicle (control) (1 mg/kg/day, i.p., 7 days); cardiac electrophysiologic parameters and atrial dimensions were determined. In adult rats, rhIL-11 was without effect, and atrial pacing did not induce AF. In contrast, in old rats, rhIL-11 shortened atrial refractoriness and increased atrial dimensions. Also, atrial pacing induced AF in old rats treated with rhIL 11 more than in controls. These effects of rhIL-11 in old rats were reversed by sodium-restricted diet. It was concluded that rhIL-11 induces age-dependent atrial remodeling manifested by atrial stretch and reduced atrial refractoriness, which favor AF. Na retention is the most likely mechanism underlying this effect of rhIL-11. PMID- 11862124 TI - A ruthenium (III) polyaminocarboxylate complex, a novel nitric oxide scavenger, enhances graft survival and decreases nitrosylated heme protein in models of acute and delayed cardiac transplant rejection. AB - Nitric oxide (NO) derived from the up-regulation of inducible NO synthase (iNOS) is believed to play an important role in organ rejection. In experimental models of acute cardiac transplant rejection (i.e., without immunosuppression), treatment using NOS inhibitors to prevent acute rejection have yielded conflicting results. This is most likely due to potential inhibition of constitutive NOS. Accordingly, agents that trap NO directly may have some advantage. In the current study, we evaluated the actions of a ruthenium-based NO scavenger, AMD6221, to inhibit the nitrosylation of myocardial protein and to prolong cardiac allograft survival in a model of acute cardiac transplant rejection (without immunosuppression). In addition, we evaluated the efficacy of AMD6221 used in combination with low-dose cyclosporine (CsA) (i.e., a model of delayed graft rejection). Heterotopic abdominal cardiac transplantation was performed using rat strains with disparities at major and minor histocompatibility loci. Grafts were harvested on postoperative day 6 for histologic examination or analysis of myocardial protein nitrosylation using electron paramagnetic resonance (EPR) spectroscopy. Other animals were monitored twice daily to determine rejection times. Plasma was also taken at postoperative day 6 for determining the concentration of NO by-products (nitrate plus nitrite). Treatment with AMD6221 either prolonged graft survival and/or caused a marked decrease in myocardial nitrosylprotein formation as determined by EPR spectroscopy. In vivo scavenging of NO by AMD6221 was verified by high performance liquid chromatography analysis of nitrosylated-drug in plasma samples. Low-dose CsA given alone or in combination with AMD6221 completely blocked formation of myocardial nitrosylprotein complexes. Whereas low-dose CsA alone prolonged graft survival, combination therapy with CsA plus AMD6221 produced a synergistic effect on graft survival. These studies indicate that treatment with a ruthenium-based NO scavenger, such as AMD6221, may be an effective regimen used alone or in combination with CsA to protect myocardial proteins from posttranscriptional modification and to prolong cardiac graft survival. PMID- 11862126 TI - Effects of the I(Kr)-blocking agent dofetilide and of the I(Ks)-blocking agent chromanol 293b on regional disparity of left ventricular repolarization in the intact canine heart. AB - Recent in vitro studies have described regional differences of ion current expression and function, possibly accounting for reduced homogeneity of repolarization in the heart. In 11 intact canine hearts regional disparity of repolarization was determined at baseline and after administration of the I(Kr) blocking agent dofetilide (30 microg/kg) and the I(Ks)-blocking agent chromanol 293b (10 mg/kg). Effective refractory periods (ERPs) were determined through up to 10 needle electrodes inserted into basal, midwall and apical regions of the left ventricular wall using the extrastimulus technique (cycle length [CL] 300 and 850 ms). At baseline (CL of 850 ms), ERPs were significantly longer in epicardial muscle layers of the apex compared to the base. In deeper muscle layers regional differences of ERPs were not detectable. Administration of dofetilide increased apico-basal disparity of repolarization, due to a more marked increase of ERPs in the apex than in the base. In contrast, homogeneous ERPs were evident along the apico-basal axis after administration of chromanol 293b. Transmural dispersion of refractoriness could not be observed in any region at baseline, or after drug-administration. In the intact canine heart, apico basal disparity of repolarization varies between individual muscle layers. Dependent on their current specificity, antiarrhythmic agents may enhance or diminish regional disparity of repolarization. PMID- 11862125 TI - Lysophosphatidylcholine is a major contributor to the synergistic effect of mildly oxidized low-density lipoprotein with endothelin-1 on vascular smooth muscle cell proliferation. AB - Endothelin-1 (ET-1) and oxidized low-density lipoprotein (ox-LDL) are associated with atherosclerosis and essential hypertension. We assessed the effect of mildly oxidized LDL (mox-LDL) and ox-LDL and their major oxidative components, i.e., reactive oxygen species (ROS), lysophosphatidylcholine (LPC), and 4-hydroxy-2 nonenal (HNE) and their interaction with ET-1 on vascular smooth muscle cell (VSMC) proliferation. Growth-arrested VSMCs isolated from the rabbit aorta were incubated with different concentrations of LDL, mox-LDL, ox-LDL, hydrogen peroxide (H(2)O(2)) (a donor of ROS), LPC, or HNE with or without ET-1. DNA synthesis in VSMCs was measured by [(3)H] thymidine incorporation. Mox-LDL, ox LDL, H(2)O(2), LPC, HNE, or ET-1 stimulated DNA synthesis in a dose-dependent manner. Maximal effect was observed at 5 microg/ml for mox-LDL (162%) or ox-LDL (154%), 15 microM LPC (156%), 5 microM H2O2 (177%), 1 microM HNE (144%), and 0.1 microM ET-1 (195%). By contrast, LDL was without any significant effect. When added together, there was no synergistic effect of LDL, H2O2, or HNE with ET-1 on DNA synthesis. However, the effect of mox-LDL (0.1 microg/ml), ox-LDL (0.5 microg/ml), or LPC (10 microM) was potentiated by ET-1 (114%-338%, 133%-425%, 118%-333%, respectively). The mitogenic effect of mox-LDL, ox-LDL, or LPC and their interaction with ET-1 were inhibited by defatted albumin (10 microg/ml), antioxidant N-acetylcysteine (400 microM), the reduced nicotinamide adenine dinucleotide phosphate (NADPH) oxidase inhibitor diphenylene iodonium (1 microM). The ET(A/B) receptor antagonist TAK044 (1 microM) or the MAPK kinase inhibitor PD098059 (10 microM) inhibited the mitogenic effect of ET-1 and its interaction with mox-LDL, ox-LDL, or LPC. The synergistic interaction of mox-LDL, ox-LDL, or LPC with ET-1 was completely reversed by the combined use of N-acetylcysteine and TAK044. Our results suggest that mox-LDL, ox-LDL, and their major phospholipid component LPC act synergistically with ET-1 in inducing VSMC proliferation by way of the activation of redox-sensitive and MAPK pathways. PMID- 11862127 TI - Sedation for peritonsillar abscess drainage in the pediatric emergency department. AB - OBJECTIVE: To evaluate the use of intravenous (IV) sedation in children during peritonsillar abscess (PTA) incision and drainage in the emergency department (ED). DESIGN: Retrospective review of medical records of children with a diagnosis of PTA. SETTING: The ED of a large, urban, academic children's hospital. PATIENTS: Consecutive patients 18 years or younger presenting from April 1995 to November 1998. METHODS: Information was retrieved from a time-based sedation record that included age, sex, ASA classification, time since last liquid or solid, agent and dose, level of sedation (A=alert, V=response to voice, P=purposeful response to pain, U=unresponsive), vital signs, complications, recovery time, and disposition. RESULTS: Forty-two patients had incision and drainage performed with IV sedation in the ED. Mean age was 11.3 +/- 4.3 years (range 4-18 years); 57% were African-American, and 64% were female. Agents used included ketamine plus midazolam (K/M) (n = 36, 86%), morphine plus midazolam (n = 3, 7%), meperidine plus midazolam (n = 2, 5%), and nitrous oxide plus midazolam (n = 1, 2%). No cardiorespiratory complications, including laryngospasm, occurred. Vomiting occurred in 1 patient who received meperidine and midazolam. The deepest level of sedation reached included: 12% A, 64% V, and 24% P. No patient who had an abscess drained in the ED with IV sedation was admitted, and mean recovery time was 81.0 +/- 30.1 minutes. CONCLUSIONS: IV sedation in children for incision and drainage of PTA by skilled personnel in the ED may eliminate the need for admission and surgical drainage in the operating room. K/M was used most frequently, without adverse effect, and all patients were discharged from the ED. Because K/M may result in deep sedation, appropriate personnel and equipment must be present. PMID- 11862128 TI - Smoking in parents of children with asthma and bronchiolitis in a pediatric emergency department. AB - OBJECTIVES: To determine smoking habits, levels of nicotine-addiction, readiness to quit, and beliefs about the effects of environmental tobacco smoke (ETS) of parents of children with bronchiolitis and asthma who present to a children's emergency department (ED). DESIGN/METHODS: This was a cross-sectional prevalence study of parents or legal guardians of children with asthma or bronchiolitis presenting to a pediatric ED. RESULTS: Two hundred forty-nine parents/legal guardians comprised the study group. The mean age (+/-SD) was 30.0 (+/-8.9) years; 88% were female; 51% were nonwhite; 37% were educated beyond high school. The self-reported smoking prevalence was 41% (95% CI = 32-51). Smoking prevalence among parents of wheezing children varied according to education, income, and race, but not according to gender, age, or employment status. Of the 102 smokers in the sample, 84 (82.4%, 95% CI = 73-88) reported that they wanted to quit; 78 (76.5%, 95% CI = 68-84) stated that they wanted to quit within the next month. Forty-nine percent (95% CI = 39-59) scored above 4 on the Fagerstrom Test for Nicotine Dependence and were considered nicotine-dependent. The majority of smokers admitted to smoking around their children (66.7%, 95% CI = 57-75). Many parents knew that ETS might contribute to the development of the following illnesses: colds/upper respiratory tract infections - 77.5%, otitis media - 68.6%, pneumonia - 50%, wheezing/asthma attacks - 86.3%, and SIDS - 31.4%. CONCLUSION: The prevalence of smoking and nicotine addiction among parents of children with asthma or bronchiolitis who bring their children to a pediatric ED is high. Many parents have some knowledge about the effects of ETS, and the majority would like to quit. Future studies to help determine the best way to deliver advice to parents on ETS exposure reduction and smoking cessation are warranted. PMID- 11862129 TI - Do clinical variables predict pathologic radiographs in the first episode of wheezing? AB - OBJECTIVE: To determine if clinical variables assessed in relation to Albuterol aerosol treatments accurately identify children with pathologic radiographs during their initial episode of bronchospasm. METHODS: A prospective convenience sample of children with a first episode of wheezing. Data collected included demographics, baseline and post-treatment clinical score and physical examination, number of aerosols, requirement for supplemental oxygen, and disposition. Chest radiographs were obtained and interpreted, and patients were divided into 2 groups based on a pathologic versus nonpathologic radiograph interpretation. Chi2 testing was performed for categoric variables, and the student t test was performed for continuous variables. A discriminant analysis was used to develop a model. RESULTS: Pathologic radiographs were identified in 61 patients (9%). Between groups, a significant difference was noted for pretreatment oxygen saturation only. Clinical score, respiratory rate, and presence of rales both pretreatment and posttreatment were not significantly different between groups. The discriminant analysis correctly predicted 90% of nonpathologic radiographs but only 15% of pathologic radiographs. CONCLUSIONS: Clinical variables, either isolated or as components of a model, could not identify all children with pathologic radiographs. PMID- 11862130 TI - Test characteristics of the urine Gram stain in infants or= 38.0 degrees C presenting to a pediatric emergency department during 2 consecutive winter seasons. Single pathogen growth of >or= 10(4) cfu/mL from a catheterized specimen and >or= 10(3) cfu/mL from a suprapubic specimen was considered positive. A positive Gram stain was defined as the identification of any organisms. Urinalysis was tested for the presence of nitrites and for leukocyte esterase (LE). Urine microscopy was analyzed for white blood cells per high power field (WBC/hpf). RESULTS: Fourteen of 246 patients were excluded; 11 because no Gram stain was completed. Of the remaining 232 patients, Gram stain had a sensitivity of 85.2% (95% CI 71.9-98.6%), a specificity of 99.0% (95% CI 97.7 100%), a likelihood ratio for a positive test of 87.3 (95% CI 21.8-349.9), and a likelihood ratio for a negative test of 0.15 (95% CI 0.06-0.37). There were 193 specimens for which a Gram stain and a complete UA and microscopy were completed and compared. Urine microscopy had a lower sensitivity and specificity than Gram stain for both >or= 5 WBC/hpf and >or= 10 WBC/hpf. In addition to the Gram stain, a dipstick negative for nitrites and LE had a low negative likelihood ratio (0.16), useful for decreasing the likelihood of a UTI. CONCLUSIONS: The Gram stain has excellent test characteristics for the preliminary diagnosis of a UTI in febrile infants. Patient disposition and therapy will likely change if clinical protocols and guidelines use the Gram stain rather than urine microscopy for preliminary diagnosis of UTI in infants. PMID- 11862131 TI - Data acquisition in emergency medicine: electronic communication using free text. AB - OBJECTIVES: 1) To describe the content of medical data obtained from parents' use of free text during a computer interview; 2) To assess whether differences in parents' demographics or experience with computers differentiate their production of free text; and 3) To compare parents' entry of free text with the final physician-generated electronic medical record (EMR) regarding the chief complaint. METHODS: Data entered by parents during a prospective trial of electronic data entry were examined. Content analysis of parents' free text entries examined absolute word count and categorized entries by type of information. Demographic and technology-related variables were analyzed against the type of information and the absolute word count. Two reviewers independently judged agreement between parents' and physicians' data for the chief complaint. RESULTS: Of 100 parents enrolled, 30 entered data for the chief complaint using free text. Amount of free text entered ranged from 1 to 142 words with a median of 8 words. Parents' free text was categorized into 4 types of information: simple descriptive (n = 16), problem-focused (n = 7), descriptive plus a question agenda (n = 2), and descriptive with reference to past medical history (n = 5). Parental demographics and computer-related experience did not predict differential production of free text. Agreement between parents' and physicians' entries for the chief complaint was 80% (95% CI 66-94) [reviewer A] and 73% (95% CI 57-89) [reviewer B]. Seven of 30 parental entries (23%) provided data not contained in the EMR. CONCLUSIONS: When entering a chief complaint during an electronic interview, a majority of parents report descriptive data using short phrases. Good agreement between parents' and physicians' electronic report was demonstrated for the chief complaint. PMID- 11862132 TI - Assessment of the palatability of vehicles for activated charcoal in pediatric volunteers. AB - OBJECTIVE: To evaluate the palatability of 4 common flavoring vehicles (water, chocolate milk [CM], orange juice [OJ], and cola) combined with activated charcoal (AC) in pediatric volunteers. DESIGN: A single-blind taste test of 4 different vehicles (water, OJ, a cola drink, and CM) was conducted in healthy volunteer children. Each child tasted 1.25 mL of Charcodote (0.2g/mL) mixed with 1.25 mL of each vehicle. SETTING: Palatability testing was conducted at the office of 1 of the authors. PARTICIPANTS: Thirty children (16 male, 14 female), aged 6.5 +/- 1.4 years (range 5-9 years). OUTCOME MEASURES: After each test dose, each child rated its taste on a modified 10 cm visual analog scale incorporating a facial-hedonic scale. Each child was also asked for his/her overall preference. RESULTS: Taste scores (cm) were as follows: water 5.6 +/- 1.8, OJ 5.4 +/- 1.0, cola 7.6 +/- 0.7, and CM 5.6 +/- 0.8. There was a significant difference in the taste scores between the cola drink (P = 0.01) and the other 3 vehicles. The cola drink was also selected as the most preferred vehicle by 50% of the children as compared with 19.2% for CM and 15.4% for OJ. In contrast, water was selected as the least preferred vehicle by 36.4% of children versus 31.8% for CM and 27.3% for OJ. Only 4 children (15.4%) stated that water was their preferred vehicle, and only 1 child (4.5%) stated that cola drink was the least preferred drink. CONCLUSIONS: Children rate the palatability higher and prefer charcoal given with a cola drink rather than with water. OJ and CM do not seem to improve the acceptability of charcoal. PMID- 11862133 TI - Diabetic ketoacidosis in a child on FK506 immunosuppression after a liver transplant. AB - The use of immunosuppressive agents is becoming more widespread, especially in the context of organ transplantation. We report a child with a complication, new onset diabetes mellitus with diabetic ketoacidosis, associated with the use of one such agent, FK506 (tacrolimus). PMID- 11862134 TI - Suicidal psychosis secondary to isoniazid. PMID- 11862135 TI - Ventriculoperitoneal shunt migration presenting with vaginal discharge and hydrosalpinx in a 16-year-old patient. PMID- 11862136 TI - Multiple splenic infarcts associated with toxic shock syndrome. PMID- 11862137 TI - Cerebrovascular event, dilated cardiomyopathy, and pheochromocytoma. AB - Cerebral infarction in children may be the result of various disease processes, including emboli from intracardiac sources, paradoxical emboli from the venous system, sickle cell disease, cyanotic heart disease, vasculitis affecting the carotid or cerebral vascular system, vascular anomalies, and prothrombotic states. We present a previously healthy adolescent who presented with the acute onset of hemiparesis. Work-up revealed a dilated cardiomyopathy with a left ventricular mural thrombus as the etiology of his cerebrovascular event. Although dilated cardiomyopathy (DCM) may predispose to the development of a mural thrombus and subsequent embolic events, there are no previous reports in pediatric-aged patients of the development of an embolic event as the presenting manifestation of DCM. Further investigation of the etiology of the DCM led to the diagnosis of a pheochromocytoma. Congestive heart failure and DCM as the presenting sign of pheochromocytoma has likewise not been reported in a pediatric aged patient. We review this unlikely sequence of events, the diagnostic evaluation of such patients, and treatment options. PMID- 11862138 TI - Hyperinsulinemia/euglycemia therapy for calcium channel blocker poisoning. PMID- 11862139 TI - The state of pediatric interfacility transport: consensus of the second National Pediatric and Neonatal Interfacility Transport Medicine Leadership Conference. AB - Interfacility transport of pediatric and neonatal patients for advanced or specialty medical care is an integral part of our medical delivery system. Assessment of current services and planning for the future are imperative. As part of this process, the American Academy of Pediatrics and the Section on Transport Medicine held the second National Pediatric and Neonatal Transport Leadership Conference in Chicago in June 2000. Ninety-nine total participants, representing 25 states and 5 international locations, debated and discussed issues relevant to the developing specialty of pediatric transport medicine. These topics included: 1) the role of the medical director, 2) benchmarking of neonatal and pediatric transport programs, 3) clinical research, 4) accreditation, 5) team configuration, 6) economics of transport medicine in health care delivery, 7) justification of transport teams in institutions, and 8) international transport/extracurricular transport opportunities. Insights and conclusions from this meeting of transport leaders are presented in the consensus statement. PMID- 11862141 TI - Closed head injury in children. PMID- 11862140 TI - A 9-month-old baby with subdural hematomas, retinal hemorrhages, and developmental delay. PMID- 11862142 TI - Care of the female adolescent rape victim. PMID- 11862143 TI - Pediatric emergency medicine: Legal briefs. PMID- 11862160 TI - Hepatic cirrhosis increases sensitivity of kidney to endotoxin in rats. AB - BACKGROUND: Renal failure in cirrhotic patients is a severe complication and endotoxemia might be involved. We investigated the effect of endotoxin on renal function of cirrhotic rats and the potential protective role of N-acetylcysteine (NAC). MATERIAL/METHODS: Hepatic cirrhosis was generated in a rat model by carbon tetrachloride. Both cirrhotic and normal rats were insulted by endotoxin intravenously, while another cirrhotic group was pre-treated with NAC. Blood urea nitrogen (BUN) and creatinine were assayed eight hours later. The changes in serum tumor necrosis factor-a (TNF-a) were assayed by ELISA. The histological changes in the kidney were observed after hematoxylin and eosin staining. RESULTS: Endotoxin increased the BUN and creatinine levels in both normal and cirrhotic rats, with a much higher elevation in the latter group. TNF-a concentration was also increased by endotoxin; the changes are positively correlated with BUN and creatinine. NAC pretreatment significantly attenuates the effects of endotoxin on BUN, creatinine and TNF-a levels in cirrhotic rats with no improvement in systemic toxicity symptoms. There were no obvious histological changes in the kidney of these animals. CONCLUSIONS: Hepatic cirrhosis increased the sensitivity of renal function to endotoxemia, which may be protected by NAC. PMID- 11862161 TI - PACAP and VIP in the photoneuroendocrine system (PNES). AB - The presence of PACAP and VIP was demonstrated in all the four levels of the photoneuroendocrine system (PNES) with the use of immunohistochemistry (IHC), radioimmunoassay (RIA), anterograde and retrograde tracing techniques, and cell immunoblot assay (CIBA). Both peptides play a physiological role in the PNES. According to our results both PACAP and VIP are involved in the regulation of the gonadotrop hormone secretion and their inhibitory role may be mediated through the neuronal chain of the PNES. PMID- 11862162 TI - Parents rate the ratings: a test of the validity of the American movie, television, and video game ratings. AB - BACKGROUND: Numerous studies have documented the potential effects on young audiences of violent content in media products, including movies, television programs, and computer and video games. Similar studies have evaluated the effects associated with sexual content and messages. Cumulatively, these effects represent a significant public health risk for increased aggressive and violent behavior, spread of sexually transmitted diseases, and pediatric pregnancy. In partial response to these risks and to public and legislative pressure, the movie, television, and gaming industries have implemented ratings systems intended to provide information about the content and appropriate audiences for different films, shows, and games. METHODS: We conducted a panel study to test the validity of the current movie, television, and video game rating systems. Participants used the KidScore media evaluation tool, which evaluates films, television shows, and video and computer games on 10 aspects, including the appropriateness of the media product for children on the basis of age. RESULTS: Results revealed that when an entertainment industry rates a product as inappropriate for children, parent raters agree that it is inappropriate for children. However, parent raters disagree with industry usage of many of the ratings designating material suitable for children of different ages. Products rated as appropriate for adolescents are of the greatest concern. The level of disagreement varies from industry to industry and even from rating to rating. CONCLUSIONS: Analysis indicates that the amount of violent content and portrayals of violence are the primary markers for disagreement between parent raters and industry ratings. Short-term and long-term recommendations are suggested. PMID- 11862163 TI - Ambulatory blood pressure monitoring in children and adolescents. Current insights into a new technique. AB - Ambulatory blood pressure monitoring (ABPM) is a new technique recently introduced into paediatrics. Devices with auscultatory blood pressure measurement may be more precise but give more erroneous readings than those with oscillometric measurement. For the interpretation of the results, mean day- and night-time values should be calculated according to patients' diaries and gender- and height-specific percentiles for ABPM measurements should be used to define hypertension. The use of "blood pressure load" as the percentage of single readings above a given upper level is not standardised in children and adolescents. A nocturnal blood pressure decline of less than 10% is defined as "non-dipping", although this is an arbitrary definition whose prognostic value is not evaluated in paediatric patients. Several studies showed that ABPM in children is superior in detecting "white-coat''-hypertension, shows a better reproducibility and is a better prognostic factor for left ventricular mass compared to casual blood pressure measurements. Clinical studies show the value of ABPM in obese children, in children with renal diseases, on dialysis therapy and after renal transplantation as well as in diabetic children or in children after surgical repair of aortic coarctation. Ambulatory blood pressure monitoring in children and adolescents has grown to a valuable method for the daily management of patients with suspected or established hypertension. However, there is still a lot of work before all technical problems are solved and international guidelines can be published which are based on evidence from follow-up studies. PMID- 11862165 TI - The role of the pediatrician in preventing suicidal behavior. AB - Suicidality is a serious cause of morbidity and mortality among young people. Important risk factors for suicidal behavior are mental illness, alcohol and other substance use disorders, previous suicide attempt, impulsive and/or aggressive behavior, history of abuse, and access to lethal means. Emotional well being and connectedness to family and school act to buffer or protect young people from involvement in self-directed violence. Pediatricians can play a major role in suicide prevention by identifying emotional and behavioral problems and intervening appropriately, promoting positive parenting skills and family cohesion, and providing injury prevention education to reduce access to lethal means. As part of a comprehensive strategy to prevent youth suicidal behavior, child health professionals are uniquely positioned to promote resiliency among youth and families as well as identify and provide appropriate treatment and service coordination for risk factors before injuries occur. Adequate training is critical to ensure that pediatricians are prepared to provide effective assessment, prevention and intervention for suicidal behavior. PMID- 11862164 TI - Improving adolescent health outcomes. AB - Adolescent morbidity and mortality are more often due to preventable causes and to risky behavioral choices than to "natural" causes, such as cardiovascular disease or cancer. The leading causes of death among adolescents in Western, industrialized nations are unintentional injuries, especially motor vehicle crashes, homicide, and suicide. The physical and cognitive development of adolescents also results in increased risky behavioral choices, and to high rates of sexually transmitted diseases, substance use and misuse, and inadequate nutrition. These lifestyle choices also have repercussions that last into adulthood. Brief counseling interventions by physicians or other clinicians have been shown to be effective in modifying health risk behaviors in adolescents. Adolescents also have indicated both a belief that physicians should counsel them on risk behaviors and a willingness to discuss risk behaviors if asked about them in a confidential manner. In this paper, we review the leading causes of adolescent morbidity and mortality in the United States and Western Europe, including injuries, violence, depression and suicide, substance use, sexual activity, and nutrition, physical activity, and eating disorders. In addition, we describe the effectiveness of physician counseling for reduction or prevention of specific risk behaviors and the importance of providing comprehensive, confidential care. Additionally, we describe the results of a recent study of implementation of Adolescent Preventive Service Guidelines in community and migrant health centers that increased risky behavior screening and counseling for adolescent patients seen for routine/well care visits. PMID- 11862166 TI - Successful surgical treatment of ventricular tachycardia that induced left ventricular aneurysm in 12-year-old boy. AB - A surgical treatment of ventricular tachycardia (VT) that induced a congenital left ventricular aneurysm (LVA) in a 12-year-old boy was performed. The VT disappeared after epicardial cryoablation and a reinforcement of LVA with a composite patch. Epicardial cryoablation, based on an intraoperative electrophysiologic study (EPS), is effective in treating VT resistant to antiarrhythmia drugs. PMID- 11862167 TI - Early-onset of septo-optic dysplasia. A case report with follow-up. AB - The case of a nine-month'old girl with septo-optic dysplasia is reported. The patient suffered in neonatal life from severe hypoglycemia, conjugated hyperbilirubinemia and nystagmus. At the age of 2 months ophthalmological examination revealed bilateral optic nerve hypoplasia; at the age of 9 months endocrinological investigations showed GH and ACTH deficiency and substitutive hormone therapy was started. MRI scans demonstrated dilatation of ventricles and pituitary ectopia. At the age of 27 months hypothyroidism due to TSH deficiency developed. The auxological and neurological follow-up is reported. The importance of direct ophthalmoscopy of the optic nerve in neonatal age is stressed: an early diagnosis can favourably influence the outcome of the disease. PMID- 11862168 TI - [Professional autonomy in the choice and management of therapy. Medicolegal aspects]. AB - Any discussion of the delicate subject of a doctor's autonomy in the choice and management of therapy entails addressing the intimate question of professional and ethical conduct. The professional freedom to choose which therapy to use continues to be the most quintessential expression of the medical professional and, in respect of the primary purpose of safeguarding health, must be aligned with: the choices of the informed patient or his/her legal representatives, existing guidelines and the equitable allocation of resources. On these grounds, the author sets out brief considerations regarding a number of issues frequently raised in the everyday work of the pediatrician and neonatologist: from the doctor's autonomy to adopt a diagnostic-therapeutic approach, with special reference to the prescription of drugs and the subtle differences compared to pharmacological trials, to the promotion and protection of breastfeeding, with explicit reference to the Ministry of Health's recent Circular no. 16 of 24.10.2000 ("Promotion and protection of breastfeeding") which, following a cursory reading of the text, appears to highlight restrictions to the pediatrician's decision-making autonomy. Special emphasis is also given to the medicolegal problems of caring for high-risk neonates. The clinical practice of neonatology often places the doctor in "critical" decision-making situations, with far from negligible ethical and moral implications, not to mention juridical and forensic ones. The author sets out a number of decision-making parameters for use as guidelines: the statistical criterion, the legal criterion and the clinical criterion, the only one capable of adjusting to scientific progress and to the specific technical and professional resources of the medical centre where the doctor is required to intervene in order to preserve even the simple "possibility" of autonomous life for the fetus. From this point of view, the so called chronological vitality and scientifically proven statistics form part of the numerous criteria available for clinical evaluation. If the clinical criterion is not met, namely if the product of conception is not vital and has never been fully alive in the biologically complete sense of the term, then the omission of therapeutic support is not particularly important in professional and juridical terms. On the contrary, it would show an obstinate persistence of therapy, which would be both ethically and professionally incorrect. Lastly, even during the final stages of life, the doctor must play a participatory role not only by providing palliative treatment but also, and above all, offering moral assistance to sanction the ineluctable end of the therapeutic alliance, that "feeling together" which is the most literal and profound meaning of the word "consent", indicating both "information" but to an even greater extent "discussion" and "empathy" with the patient and his/her family. PMID- 11862169 TI - Practical evaluation and management of cutaneous lymphoma. AB - Accurate evaluation of patients with suspected or known cutaneous lymphoma requires the integration of many sources and types of information, including clinical evaluation, microscopic analysis of tissue, immunophenotyping, gene rearrangement studies, clinical staging, and longitudinal observation. Diagnoses should be based on knowledge of specific lymphoma types as described in modern classification systems. Management of patients with cutaneous lymphoma requires collaboration among dermatologists, dermatopathologists, hematopathologists, and medical, surgical and radiation oncologists. (J Am Acad Dermatol 2002;46:325-57.) LEARNING OBJECTIVE: At the conclusion of this learning activity, participants should better understand how to evaluate and manage patients for suspected or established lymphoma of the skin. Components include the clinical history and physical examination, optimal biopsy and tissue handling, interpretation of pathology and adjunctive test results, clinicopathologic correlation, and therapy. Participants should also understand the basis for establishing a specific diagnosis of cutaneous lymphoma based on current classification and staging. PMID- 11862170 TI - Cost of atopic dermatitis and eczema in the United States. AB - BACKGROUND: Atopic dermatitis/eczema (AD/E) is a common disease. Few studies have attempted to quantify the cost to third-party payers. OBJECTIVE: Our purpose was to identify the annual cost of medical services and prescription drugs for the treatment of AD/E to private insurance and Medicaid payers in the United States. METHODS: We used a retrospective study design employing claims data from 1997 and 1998 from a private insurer and a state Medicaid program to analyze costs incurred. Beneficiaries were considered to have AD/E if they had at least one claim in 1997 with a primary or secondary listing of 1 of 3 diagnosis codes: 691.8, other atopic dermatitis and related conditions; 692.9, contact dermatitis and other eczema when no cause is specified; or 373.3, noninfectious dermatoses of eyelid. Patients who did not meet the diagnosis criteria served as a control group in each payer for comparisons of expenditures with the AD/E group. RESULTS: Disease prevalence was 2.4% (private insurer) to 2.6% (Medicaid) of all eligible beneficiaries, and 3.5% to 4.1% of patients submitted at least one health care claim during the study period. Medicaid-insured patients used outpatient hospital visits and hospitalizations at a greater rate than did privately insured patients; neither used emergency departments extensively. The third-party payer cost of illness for AD/E ranged from $0.9 billion to $3.8 billion when projected across the total number of persons younger than 65 years insured by private insurers and Medicaid in the United States. More than one fourth of all health care costs for patients with AD/E may be attributed to AD/E and co-morbid conditions. CONCLUSIONS: Annual costs of AD/E are similar to those of other diseases such as emphysema, psoriasis, and epilepsy. Patients incur significant costs associated with AD/E and co-morbid conditions. PMID- 11862171 TI - Reactivity to trichophytin antigen in patients with onychomycosis: effect of terbinafine. AB - BACKGROUND: Many patients with chronic dermatophytosis and onychomycosis have depressed cell-mediated immunity (CMI) to trichophytin. OBJECTIVE: The fungicidal properties of oral terbinafine provide a unique opportunity to explore whether elimination of antigen could restore CMI response in these patients. METHODS: A double-blind, placebo-controlled study evaluated the effect of terbinafine (250 mg/d for 12 weeks) on skin immunoreactivity to intradermal trichophytin antigen (TRIPA), mycologic status of the nail, and nail growth in patients with toenail onychomycosis. RESULTS: Skin reactivity, in an optimized, dose response challenge series to TRIPA was inversely related to disease chronicity. Mycologic/clinical response rates were 72%/84% for terbinafine and 0%/7% for placebo. Terbinafine increased the number of TRIPA reactors 2-fold and the mean TRIPA reaction area 4 fold; responses in placebo-treated patients were relatively unchanged. Of the 7 (of 25) patients receiving terbinafine who still had positive mycology 6 months after treatment, all were anergic to TRIPA at baseline and all but one remained so after treatment. CONCLUSION: Terbinafine treatment enhances and restores CMI to TRIPA in patients with Trichophyton rubrum onychomycosis and may thereby reduce susceptibility to reinfection. Terbinafine reversal of immunologic anergy may be an important model of microbial tolerance in chronic dermatophyte infections. PMID- 11862173 TI - Venous leg ulcer: incidence and prevalence in the elderly. AB - BACKGROUND: Venous leg ulcer is a disease most common in those aged 65 years and older. However, the incidence and prevalence have not been well established. OBJECTIVE: Our purpose was to estimate the prevalence and incidence of venous leg ulcers in the elderly. METHODS: We studied the General Practice Research Database, validated our case ascertainment strategy, and estimated the annual prevalence and incidence of venous leg ulcer. RESULTS: The positive predictive value of our ascertainment strategy was 98.3% (95% confidence interval [CI], 90.0, 100.0). The annual prevalence of venous leg ulcer among the elderly was 1.69 (95% CI, 1.65, 1.74). The overall incidence rate was 0.76 (95% CI, 0.71, 0.83) for men and 1.42 (1.35, 1.48) per 100 person-years for women. CONCLUSION: Venous leg ulcer is a significant problem in those aged 65 years and older. As compared with previous studies, because we used medical records from a population of known size, our results are precise and less biased. PMID- 11862172 TI - Tuberous sclerosis-associated lesions of the kidney, brain, and skin are angiogenic neoplasms. AB - BACKGROUND: Tuberous sclerosis is an autosomal dominant condition characterized by the development of benign neoplasms of the brain, kidney, and skin. Progressive growth and malignant transformation of brain and kidney lesions constitute the major cause of morbidity and mortality in adults with tuberous sclerosis. In addition, growth of skin lesions may be disfiguring to patients. OBJECTIVE: The purpose of this study was to determine whether benign tumors in patients with tuberous sclerosis are angiogenic. METHODS: Brain, kidney, and skin tumors from patients with tuberous sclerosis were stained with CD31, a specific marker of vascular endothelium. In addition, we used Northern blot analysis to demonstrate that renal angiomyolipoma cells express the potent angiogenesis stimulator vascular endothelial growth factor (VEGF). RESULTS: Brain, kidney, and skin neoplasms from patients with tuberous sclerosis are highly angiogenic. Renal angiomyolipoma cells produce the potent angiogenic factor VEGF. CONCLUSION: Benign neoplasms of patients with tuberous sclerosis are highly vascular. Our results provide a rationale for antiangiogenic therapy in the treatment and prevention of tuberous sclerosis-associated neoplasms. PMID- 11862174 TI - Safety of fluticasone propionate cream 0.05% for the treatment of severe and extensive atopic dermatitis in children as young as 3 months. AB - BACKGROUND: Topical corticosteroids are useful for the treatment of pediatric dermatoses. However, concerns regarding possible systemic and topical toxicities have limited the use of moderate-potency corticosteroids in children. OBJECTIVE: Our purpose was to characterize the safety of fluticasone propionate cream in children. METHODS: Children between 3 months and 5 years 11 months (n = 32) and 3 up to 6 years of age (n = 19) with moderate to severe atopic dermatitis (> or =35% body surface area; mean body surface area treated, 64%) were treated with fluticasone propionate cream, 0.05% twice daily for 3 to 4 weeks. Serum cortisol response, fluticasone levels, skin changes, and adverse events were analyzed. RESULTS: Mean cortisol levels were similar at baseline (13.76 +/- 6.94 microg/dL prestimulation and 30.53 +/- 7.23 microg/dL poststimulation) and at end of treatment (12.32 +/- 6.92 microg/dL prestimulation and 28.84 +/- 7.16 microg/dL poststimulation). Only 2 of 43 children had end-treatment poststimulation values less than 18.0 microg/dL. No significant adverse cutaneous effects were noted. CONCLUSION: Fluticasone propionate cream 0.05% appears to be safe for the treatment of severe eczema for up to 4 weeks in children 3 months of age and older. PMID- 11862175 TI - Subungual myxoid cysts: clinical manifestations and response to therapy. AB - BACKGROUND: Myxoid cysts located beneath the nail are very different from those within the nailfold and there is no established literature on their diagnosis and therapy. OBJECTIVE: Thirty-five cases of subungual myxoid cyst are described in which the diagnosis was confirmed by magnetic resonance imaging, surgical exploration, or both. METHODS: The clinical characteristics of all cases of subungual myxoid cysts confirmed in 3 specialist nail clinics are reported. Twenty cases were confirmed by magnetic resonance imaging. Histologic details were recorded in 4 cases. RESULTS: Thirty-four patients (22 female, 12 male) presented with a subungual tumor of the thumb (n = 25) or finger (n = 10). The 3 main clinical variables were color of the lunula, transverse nail curvature, and integrity of the nail. In 26 cases the lunula was discolored; it was red in 22 cases, blue in 3, and as part of a longitudinal leukonychia in one. Transverse curvature was increased in 29 cases and was normal in 5 cases; in one case the nail was destroyed. In 11 cases, the change in curvature led to lateral ingrowing. Surgery (n = 22), sclerosant (n = 1), and spontaneous discharge (n = 2) all resulted in resolution. Ten patients had no treatment. CONCLUSION: This study describes 35 subungual myxoid cysts having characteristic features that allow clinical diagnosis. They respond to a range of treatments. PMID- 11862176 TI - Optimizing wound healing in the face after laser abrasion. AB - BACKGROUND: Laser resurfacing is a popular procedure to improve the physical signs of photoaging. In addition to improvements in treatment modalities, optimizing posttreatment regimens will enhance patient care. OBJECTIVE: Our purpose was to evaluate the efficacy of two forms of wound care for the face after laser abrasion. METHODS: Forty-two patients received full-face laser resurfacing at two clinics by using either the UltraPulse carbon dioxide (CO(2)) laser (Coherent Laser Corp, Palo Alto, Calif) alone or followed by an erbium:YAG laser (Derma-20, ESC Sharplan, Inc, Needham, Mass) and/or a blended CO(2)/Er:YAG laser (Derma-K, ESC Sharplan) or a variable pulse erbium:YAG laser (Contour, Sciton Laser Corp, Palo Alto). Twenty-one patients were randomly assigned to a postoperative regimen including Silon-TSR (Bio Med Sciences, Inc, Allentown, Pa) for the first 2 to 3 days after laser resurfacing, followed by Aquaphor ointment (Beiersdorf, Charlotte, NC) to complete the first 2 weeks. The other 21 patients received the resurfacing recovery system (RRS, Neutrogena, Los Angeles, Calif) following a specific regimen. The system includes Fibracol wound dressing (Johnson & Johnson, Skillman, NJ) for 2 days, followed by a hydrogel dressing for 1 to 2 days, followed by an ointment to complete the first 2 weeks. Patients were evaluated for wound healing on days 2, 3, 6-10, 14-16, and 28-30. The skin was swabbed for colonization at every visit to determine the quantity of bacteria throughout the healing process. RESULTS: Ninety percent of patients in both groups experienced either "no pain" or "minimal pain" during the first 3 days. Total bacterial counts peaked on days 3 and 6 in the patients managed with the RRS and the Silon-TSR/Aquaphor regimen, respectively. The average day at which patients did not require a dressing was 3.0 days in the group managed with the RRS and 3.7 days in the group managed with the Silon-TSR/Aquaphor dressing regimen (P < or =.05). The average day of complete epithelial regeneration was significantly shorter at 6.3 days using the RRS compared with 7.4 days for patients using the Silon-TSR/Aquaphor regimen (P < or =.02). There was no difference in infection, adverse sequelae, exudate management, or pain in either group. CONCLUSION: Healing was optimized in patients using the RRS after laser resurfacing. PMID- 11862177 TI - Stromelysin 3 expression: a useful marker for the differential diagnosis dermatofibroma versus dermatofibrosarcoma protuberans. AB - BACKGROUND: Stromelysin 3 (ST3) is a member of the metalloproteinase family, which is expressed in tissue remodeling processes such as scarring, embryogenesis, or tumoral invasion. Although the prognosis of breast cancers and extracutaneous squamous cell carcinomas is correlated with the level of expression of ST3, this staining has not yet found a routine application in dermatopathology. OBJECTIVE: Our purpose was to study by immunohistochemistry the expression of ST3 in dermatofibromas and dermatofibrosarcoma protuberans (DFP). METHODS: We selected 40 cases of dermatofibromas, 40 histologically typical DFPs, and 10 giant dermatofibromas. Immunohistochemistry was carried out by means of the LSAB method, with monoclonal anti-ST3 antibody (provided by MC Rio, IGBMC Strasbourg). A semiquantitative scale (0-3) was used to evaluate the level of ST3 expression. RESULTS: Positively stained cells were observed in all cases of dermatofibromas (100%), including the 10 giant cases, but never in DFP (0%). The staining was intense (class 2 or 3) in 39 of the 50 dermatofibromas. The CD34 staining used as a control proved to be less efficient; 6 DFP were CD34 negative, whereas some of the dermatofibromas showed a marginal CD34 positivity. CONCLUSION: Our results are consistent with those obtained by in situ hybridization in previous studies of smaller series of fibrous tumors. The study of ST3 expression in fibrous tumors of the skin shows that this immunostaining could be a useful tool in the purpose of differentiating DFP from giant or invasive dermatofibromas. Although ST3 is a "negative" marker for DFP and therefore does not demonstrate the margins of the neoplasm, it is more reliable than CD34 staining in differentiating this tumor from a dermatofibroma. PMID- 11862178 TI - Nevoid hyperkeratosis of the nipple and areola: a distinct entity. AB - Although nevoid hyperkeratosis of the nipple and areola was initially described in 1923, there are only case reports or reviews about it; no large series have been documented to date. The clinical features of the reported cases in the literature are not uniform, and it is questioned whether nevoid hyperkeratosis of the nipple and areola is a distinct clinicopathologic entity or a clinical presentation of various dermatoses. We describe 7 cases with hyperkeratotic nevoid lesions localized on the nipple and areola with different clinical features. None of them had any other associated dermatologic or systemic disease. Histopathologic examination was performed in 6 patients. Four of them had common histopathologic features suggesting a distinct entity, namely, nevoid hyperkeratosis of the nipple and areola; 2 of them had histopathologic features consistent with seborrheic keratosis. Seborrheic keratosis presents as sharply demarcated papules or plaques, whereas nevoid hyperkeratosis of the nipple or areola presents as a plaque diffusely involving the nipple or the areola. PMID- 11862180 TI - Surgical Pearl: Mini-running or "X" suture for closure of punch wounds. PMID- 11862179 TI - Nonendemic pemphigus foliaceus in children. AB - BACKGROUND: Pemphigus foliaceus is a cutaneous, autoimmune, blistering disease comprising two major categories: endemic and sporadic. The endemic form, also known as fogo selvagem, primarily affects children and young adults in rural Brazil. In contrast, the sporadic form of pemphigus foliaceus is generally a disease of the middle-aged and elderly. OBJECTIVE AND METHODS: Because the sporadic form of pemphigus foliaceus rarely affects children, information specific to this unique group is lacking. We describe a 3-year-old boy with the disease and retrospectively review data from 28 past cases. RESULTS: In comparison to pediatric cases of pemphigus vulgaris, sporadic pemphigus foliaceus in children tends to follow a generally benign course of relatively short duration. However, long-term outcome studies are lacking. A pattern of skin lesions described as "arcuate," "circinate," or "polycyclic" appears to be a unique and specific presentation of this disease in children. Occasionally, as in our case, the diagnosis may prove difficult to establish by using routine histology or immunopathology. CONCLUSION: The commercial availability of antigen specific techniques such as enzyme-linked immunosorbent assay for serum desmoglein 1 autoantibody should eliminate delay in diagnosis. Hydroxychloroquine may be another treatment option for those children with photodistributed lesions. Further experience and long-term outcome studies in children are needed to determine whether some medication side effects may outweigh the risks from the disease itself. PMID- 11862181 TI - Resolution of patch-type granuloma annulare lesions after biopsy. AB - We describe a patient with patch-type granuloma annulare whose lesions resolved after biopsy on 2 occasions. The lesions not subjected to biopsy persisted. There is a paucity of literature on the relation between biopsy and resolution of granuloma annulare, with one frequently cited article implying that biopsy is not related to resolution. We briefly consider possible mechanisms through which involution of lesions of granuloma annulare could result after biopsy or other form of trauma. PMID- 11862182 TI - Dengue hemorrhagic fever in a British travel guide. AB - A 44-year-old female British travel guide suddenly had fever, nausea, vomiting, and diarrhea develop during her stay in South India. Four days later she was transported to our hospital. At admission she had a high temperature, impaired respiration, and abdominal pain. Clinical examination revealed bilateral pleural effusion, hepatomegaly, and ascites. Two days later the patient showed a generalized macular rash with a conspicuous sparing of small islands of normal skin. Hemorrhagic erythema on the palms and soles as well as focal petechiae on the hard palate and scleral and conjunctival bleeding were also observed. Hypotension and renal insufficiency developed 1 week after the illness started. Laboratory investigations revealed highly elevated levels of hepatic enzymes, severe hemolytic anemia, decreased platelet counts, and abnormal coagulation values. The presumptive clinical diagnosis of dengue hemorrhagic fever was supported by serologic testing that disclosed sustained high titers of hemagglutination inhibition antibodies. Symptomatic therapy with substitution of volume and albumin, blood transfusions, and administration of antipyretics resulted in complete recovery within 6 weeks. PMID- 11862183 TI - Progressive generalized alopecia due to systemic amyloidosis. AB - We describe the case of a patient with systemic amyloidosis that manifested as generalized alopecia. Amyloidosis was documented by means of skin biopsy and urine immunoelectrophoresis. PMID- 11862184 TI - Can a spinal cord tumor cause brachioradial pruritus? AB - A woman had burning pruritus on both arms and thenar regions for 1 year. The location of pruritus was consistent with C5-C6 dermatomes. Brachioradial pruritus caused by spinal cord tumor was diagnosed. Although cases of brachioradial pruritus were reported in cervical osteoarthritis and cervical rib previously, to our knowledge, our patient is the first reported case of brachioradial pruritus caused by a tumor. PMID- 11862185 TI - CD20-negative relapse of cutaneous B-cell lymphoma after anti-CD20 monoclonal antibody therapy. AB - Despite cutaneous B-cell lymphoma often having a relatively indolent course and low mortality, it is often resistant to conventional therapy and frequently relapses. We describe a patient with widespread cutaneous B-cell lymphoma who was treated successfully with a recently approved chimeric monoclonal antibody directed against the CD20 antigen (rituximab) and the CD20-negative relapse that resulted. PMID- 11862186 TI - Hyperhidrosis in pediatric spinal cord injury: a case report and gabapentin therapy. AB - Hyperhidrosis is a relatively common condition with a multitude of causes. Spinal cord injury may be complicated by hyperhidrosis. Many forms of therapy have been reported for this phenomenon but few have been demonstrated to be efficacious. We describe a case of a child with hyperhidrosis from a spinal cord injury and report the first therapeutic use, to our knowledge, of gabapentin for hyperhidrosis. PMID- 11862187 TI - Exclusion of COL7A1 mutation in Kindler syndrome. AB - We describe a patient with Kindler syndrome with an 18-year follow-up who was initially misdiagnosed as suffering from dystrophic epidermolysis bullosa. The patient's skin showed broad reticulate labeling for collagen VII and reduplication of the lamina densa. Screening of this patient's DNA excluded any pathogenic COL7A1 mutations. PMID- 11862188 TI - Cutaneous natural killer/T-cell lymphoma. AB - Lymphomas are classified as either Hodgkin's or non-Hodgkin's. The 2 subtypes of non-Hodgkin's lymphoma that can present primarily in the skin are cutaneous T cell lymphoma and cutaneous B-cell lymphoma, both of which tend to be low-grade malignant neoplasms. Recently another distinct subtype of lymphoma was discovered, the natural killer (NK)/T-cell lymphoma, which can involve the skin in a primary or secondary fashion. The NK/T-cell subtype of lymphoma is characterized by the expression of the NK-cell antigen CD56. These CD56(+) lymphomas are further subdivided into nasal NK/T-cell lymphomas that commonly present as midfacial destructive disease and non-nasal NK/T-cell lymphomas that often arise in extranodal locations, including the skin. We report a case of aggressive NK-cell leukemia/lymphoma with numerous secondary cutaneous lesions and review the clinical and histopathologic spectrum of non-nasal CD56(+) lymphomas, with an emphasis on the dermatologic findings. PMID- 11862189 TI - The incidence of nonattendance at an urgent care dermatology clinic. AB - Nonattendance in dermatology clinics remains a burden on clinic efficiency. Nonattendance rates have been measured in an urban university-based dermatology clinic and are significant. The nonattendance rate in a dermatology clinic dedicated primarily to new acute problems is not known. We hypothesized that the nonattendance rate would be minimal in an urgent care setting. We measured the incidence of nonattendance in a university-based, urgent care dermatology clinic. The clinic is held once weekly for patients with new skin problems. Almost all patients seen in the urgent care clinic are first-time patients. We tabulated clinic attendees and nonattendance, including same-day cancellations, for 508 consecutive patients seen over a 10-month period at the urgent care clinic. We then determined nonattendance rates for the group and by payer type. The total nonattendance rate was 23.9%. The percentage of patients who failed to appear was 21.9 and same-day cancellation was 2.0. The highest nonattendance and same-day cancellation rates were noted in HMO-based programs. The lowest rate of nonattendance was in the Medicare-based program. The nonattendance rate in a dermatology clinic dedicated to new skin problems was similar to that reported in a general dermatology clinic and correlated with payer type. In addition to factors identified in previous studies that affect clinic attendance, the rate in this study may reflect spontaneous clearing of patient-identified skin problems. PMID- 11862190 TI - Environmental nickel pollution: does it protect against nickel allergy? AB - The Norwegian-Russian border area is polluted because of emissions from two local Russian nickel refineries. We patch tested the general adult population on both sides of the border. Only 7.2% of the Russian women were sensitized to nickel, compared with 27.5% of the Norwegian women. We suggest that long-term exposure to nickel may have induced immunologic tolerance in the Russian population. PMID- 11862191 TI - Topical enzyme therapy for skin diseases? PMID- 11862193 TI - Dermatitis herpetiformis in association with lichen planopilaris. PMID- 11862192 TI - Fixed drug eruption due to fluconazole: a third case. PMID- 11862194 TI - Diltiazem-induced hyperpigmentation in an African American woman. PMID- 11862195 TI - Toxic epidermal necrolysis treated with N-acetylcysteine. PMID- 11862196 TI - Treatment of Bowen's disease of the penis with imiquimod. PMID- 11862197 TI - Human papillomavirus. PMID- 11862199 TI - Long-term results of different condylotomy designs for the management of temporomandibular joint disorders. AB - OBJECTIVE: The purpose of this study was to compare the long-term results of the condylotomy techniques. STUDY DESIGN: Twenty-two patients (mean age, 20.8 years; occlusion: Class I in Angle's classification of malocclusion) were studied. All showed Wilkes stage II or early stage III. The Visual Analogue Scale (VAS), maximum mouth opening (MMO), and the positional change of the condylar segment were recorded preoperatively and postoperatively. The difference in each criterion according to the operative techniques was evaluated by means of a 1-way analysis of variance, and the difference between the preoperative value and the value in the long-term follow-up was evaluated by means of a paired t test. RESULTS: Six patients underwent an extraoral vertical ramus osteotomy (EVRO), 6 patients underwent a sagittal split ramus osteotomy (SSRO), and 10 patients underwent an intraoral vertico-sagittal split ramus osteotomy (IVSRO). The preoperative value of the maximum mouth opening (MMO) was 33.0 +/- 8.3 mm, 46.1 +/- 7.0 mm, or 40.0 +/- 7.4 mm for patients undergoing EVRO, IVSRO, or SSRO, respectively. In the long-term follow-up period, the MMO was 49.3 +/- 14.6 mm, 47.3 +/- 3.2 mm, or 48.7 +/- 5.1 mm for patients undergoing EVRO, IVSRO, or SSRO, respectively. There were no differences in the amount of the MMO among the operative techniques (P >.05). The preoperative VAS in the operated-on joints was 3.9 +/- 2.4, 5.0 +/- 1.6, or 4.7 +/- 1.4 for patients undergoing EVRO, IVSRO, or SSRO, respectively. In the long-term follow-up period, it was 1.4 +/- 2.2, 2.5 plus minus 2.0, or 3.7 +/- 1.7 for patients undergoing EVRO, IVSRO, or SSRO, respectively. There were no differences in the VAS among the operative techniques (P >.05). When each measurement preoperation was compared with the long-term follow-up, the difference was statistically significant (P =.018 in the MMO, P =.004 in the VAS). CONCLUSION: The curative effect of a condylotomy on the internal derangement of the temporomandibular joint was acceptable in the long term follow-up, but the osteotomy procedure used may be only a minor contributing factor to the long-term results. PMID- 11862198 TI - Selective embolization for head and neck vasculopathies. AB - INTRODUCTION: Selective, transarterial embolization of vascular lesions through angiographic catheters, which in superficial vasculopathies is often accompanied by intralesional local administration of sclerosing agents, has been used for more than 65 years. However, the reported experience with this therapeutic modality for head and neck lesions is rather limited. METHODS AND MATERIALS: In the current study we describe our experience with all 12 patients having major head and neck vasculature pathologies who were admitted and treated by selective embolization in our institution during a 5-year period. RESULTS: Eight patients had congenital malformations, and 4 had acquired lesions induced by trauma, irradiation, or anticoagulant therapy. Four cases were potentially life threatening because of severe blood loss and were thus treated immediately. In only 3 cases was subsequent surgical resection of the lesion necessary. The therapy administered is reported, and its outcome is discussed in respect to the available updated literature. CONCLUSIONS: The reported high rate of success with no complications may be the result of careful techniques and appropriate preoperative imaging, as well as the intraoperative and perioperative treatment with steroids and the multiple intralesional puncture sessions administered as necessary. PMID- 11862200 TI - Eating disorders: dental implications. AB - This article presents updated information on the 2 major eating disorders, anorexia nervosa and bulimia nervosa. Both conditions are found primarily in women. The eating disorders have significant morbidity and mortality associated with them. Patients are vulnerable to sudden death from cardiac arrhythmias. Suicide is a concern in some patients. The etiology of the eating disorders is unknown, but genetic, cultural, and psychiatric factors appear to play a role. Medical management may involve hospitalization to stabilize the patient, behavior modification, drugs, and psychotherapy. The long-term outcome of treatment is unclear at this time. The role of the dentist as a "case finder" is discussed. Also, the role of the dentist in restoring the dental and oral tissues to a healthy state in patients with eating disorders is presented. PMID- 11862201 TI - Reported latex allergy in dental patients. AB - OBJECTIVE: The purpose of this study was to alert doctors of dental surgery to the possibility of latex sensitivity in both outpatients and inpatients. STUDY DESIGN: The study involved 2 groups: group A was composed of 21 subjects with a history of immediate reaction in dental environment; group B was composed of 24 healthy individuals. Patients underwent skin prick tests with common inhalant allergens, with latex cross-reacting foods, with a commercial extract of non ammoniated latex, and the incremental challenge test with local anesthetics. Specific IgE to latex and to latex cross-reacting foods were measured with the fluorescent enzyme immunoassay. RESULTS: All patients in group A and none in group B were latex-allergic. Subjects who were latex-allergic were significantly more likely to be atopic and had positive IgE test to cross-reactive foods. CONCLUSIONS: Dentists and people working in a dental surgery environment must obtain detailed patient history to help identify individuals at risk of latex allergy or those actually allergic to latex. If an allergy exists, equipment used should be made of alternative materials. PMID- 11862202 TI - Relationship between salivary flow rates and Candida counts in subjects with xerostomia. AB - OBJECTIVE: This study evaluated the relationship between salivary flow and Candida colony counts in the saliva of patients with xerostomia. STUDY DESIGN: Sialometry and Candida colony-forming unit (CFU) counts were taken from 112 subjects who reported xerostomia in a questionnaire. Chewing-stimulated whole saliva was collected and streaked in Candida plates and counted in 72 hours. Species identification was accomplished under standard methods. RESULTS: There was a significant inverse relationship between salivary flow and Candida CFU counts (P =.007) when subjects with high colony counts were analyzed (cutoff point of 400 or greater CFU/mL). In addition, the median sialometry of men was significantly greater than that of women (P =.003), even after controlling for confounding variables like underlying disease and medications. Sjogren's syndrome was associated with low salivary flow rate (P =.007). There was no relationship between the median Candida CFU counts and gender or age. There was a high frequency (28%) of mixed colonization. Candida albicans was the most frequent species, followed by C parapsilosis, C tropicalis, and C krusei. CONCLUSIONS: In subjects with high Candida CFU counts there was an inverse relationship between salivary flow and Candida CFU counts. PMID- 11862203 TI - Treatment of oral candidiasis with methylene blue-mediated photodynamic therapy in an immunodeficient murine model. AB - OBJECTIVES: The purpose of this study was to evaluate the efficacy of using methylene blue (MB)-mediated photodynamic therapy to treat oral candidiasis in an immunosuppressed murine model, mimicking what is found in human patients. STUDY DESIGN: Seventy-five experimental mice with severe combined immunodeficiency disease were inoculated orally with Candida albicans by swab 3 times a week for a 4-week period. On treatment day, mice were cultured for baseline fungal growth and received a topical oral cavity administration of 0.05 mL MB solution at one of the following concentrations: 250, 275, 300, 350, 400, 450, or 500 microgram/mL. After 10 minutes the mice were recultured and underwent light activation with 664 nm of diode laser light with a cylindrical diffuser. After photodynamic therapy the mice were cultured again for colony-forming units per milliliter and then killed, their tissue harvested for histopathology. RESULTS AND CONCLUSIONS: The results indicate an MB dose-dependent effect. Concentrations from 250 to 400 microgram/mL reduced fungal growth but did not eliminate Candida albicans. MB concentrations of 450 and 500 microgram/mL totally eradicated Candida albicans from the oral cavity, resulting in reductions from 2.5 log(10) and 2.74 log(10) to 0, respectively. These results suggest that MB-mediated photodynamic therapy can potentially be used to treat oral candidiasis in immunodeficient patients. PMID- 11862204 TI - Intralesional corticosteroids as an alternative treatment for central giant cell granuloma. AB - Four cases of central giant cell granuloma were treated with intralesional infiltration of a solution of Kenacort-A (10 mg/mL, triamcinolone aqueous suspension SQUIBB) and either (1) Lidocaine 2% with epinephrine 1:200,000 Marcaine or (2) Bupivacaine, 50% mixture by volume. These cases were originally diagnosed by radiographic and histologic studies in 3 Guatemalan males--ages 31, 34, and 6 years old--and a 21/2-year-old Guatemalan girl. The average dosage of the aforementioned solution was 6 mL (equivalent to 30 mg of triamcinolone) for the adults and 5 mL (equivalent to 25 mg of triamcinolone) for the pediatric patients. Before treatment, an endocrinologist evaluated all of the patients to rule out hyperparathyroidism. Also before treatment, an incisional biopsy of the lesion was obtained from each patient for microscopic examination. Follow-up radiographs for all the cases showed progressive improvement and eventual resolution of the lesions. PMID- 11862205 TI - Tender mandibular swelling of short duration. PMID- 11862206 TI - Spindle cell lipoma of the alveolar mucosa: a case report. AB - Spindle cell lipoma of the oral cavity is extremely rare. We report a case occurring on the alveolar mucosa, an oral site that has not been previously reported. A short review of the literature is also presented. The importance of distinguishing this benign lesion from a sarcoma is highlighted. The lesion was excised, and no recurrence has been reported after 2 years. PMID- 11862207 TI - Patterns of microbial colonization in primary root canal infections. AB - OBJECTIVE: The purpose of this study was to investigate the patterns of microbial infection of root canals in untreated teeth associated with chronic periradicular lesions by use of scanning electron microscopy. STUDY DESIGN: Fifteen extracted teeth with extensive carious lesions, radiolucent lesions of varying sizes, and attached periradicular lesions after extraction were selected for study. After fixation, lesions were removed and the teeth were split into 2 halves. The teeth were then dehydrated, sputter-coated with gold, and then examined for the patterns of microbial colonization of the root canal system by using a scanning electron microscope. RESULTS: All examined root canals were infected, and bacterial cells were seen in practically all areas of the root canal system. The pattern of colonization was not uniform between specimens and even within the same specimen. Most of the root canal walls of all specimens were heavily colonized by a root canal microbiota consisting of cocci and/or rods, often forming mixed communities. Spirilla were occasionally observed as single cells or as small clusters between other bacterial forms. Bacteria were often observed penetrating the dentinal tubules. Although a shallow penetration was the most common finding, bacterial cells could be observed reaching approximately 300 microm in some specimens. Yeastlike cells were observed in 1 specimen together with bacteria. CONCLUSIONS: The root canals of teeth associated with periradicular lesions were heavily infected by bacteria and occasionally by fungi. The pattern of colonization of the root canal microbiota often showed the characteristic of a climax community, which may require special considerations regarding its elimination and prevention of clinical problems. In addition, the observed propagation of the infection to the entire root canal system in teeth associated with periradicular lesions suggests that proper therapeutic measures may be necessary to eliminate the root canal infection completely. PMID- 11862208 TI - Fusobacterium nucleatum in endodontic flare-ups. AB - OBJECTIVE: The extent to which Fusobacterium nucleatum is recovered from root canals of teeth that present with an interappointment flare-up following endodontic instrumentation was investigated. STUDY DESIGN: Included in the study were 28 patients that sought emergency treatment after initiation of root canal therapy. Only non-painful teeth that had been treated because of a necrotic pulp and periapical inflammatory lesion were studied. Root canal samples for bacterial analysis were taken, transported to a bacteriological laboratory, and processed for a semiquantitative assessment of bacterial isolates. Bacterial findings were correlated with self-assessed pain intensity as recorded by means of a Visual Analogue Scale. Clinical presentation of swelling and presence of exudate in the treated root canals were also linked. RESULTS: Bacteria were recovered from all teeth examined. Gram-negative anaerobic coccoid rods (Prevotella species and Porphyromonas species) were frequent isolates. All teeth in patients who were reported to be in severe pain (Visual Analogue Scale > or = 6) displayed F nucleatum. Nine out of 10 of these teeth also had swelling and exudate in the root canals. Samples from the remaining patients that had teeth with less pain score showed a variable bacterial recovery. None of these teeth displayed F nucleatum. CONCLUSION: F nucleatum appears to be associated with the development of the most severe forms of interappointment endodontic flare-ups. PMID- 11862209 TI - Influence of root canal dressings and sealers on repair of apical periodontitis after endodontic treatment. AB - OBJECTIVE: The objective of this study was to evaluate the histopathologic response of periapical tissues after root canal treatment of necrotic dog teeth with chronic apical periodontitis by using 2 calcium hydroxide-based root canal dressings and 2 root canal sealers. STUDY DESIGN: Seventy-eight root canals were instrumented by using 5.25% sodium hypochlorite as the irrigating solution, after which a calcium hydroxide paste (Calen/PMCC or Calasept) was placed for 30 days as a dressing. The root canals were then filled by using cold lateral gutta percha condensation and an endodontic sealer (Sealapex or AH Plus). After 360 days, the animals were killed by anesthetic overdose; then, the teeth were histologically prepared, sectioned, and stained with hematoxylin and eosin for optical microscopic analysis of apical and periapical tissue repair. RESULTS: Statistical analysis showed that the poorest histopathologic results were observed in the Calasept/AH Plus group and that the Sealapex sealer overall resulted in better apical repair than the AH Plus sealer. The histopathologic results of Calen/PMCC paste with both AH Plus and Sealapex and Calasept paste with only Sealapex were statistically similar but were different from the results of Calasept with AH Plus. CONCLUSIONS: The results of this study in the dog showed differences in apical and periapical tissue repair of teeth with chronic apical periodontitis by using 2 calcium hydroxide root canal dressings and 2 sealers. More research is necessary to determine the best combination of dressings and sealers. PMID- 11862210 TI - The influence of dental operating microscope in locating the mesiolingual canal orifice. AB - OBJECTIVE: The purpose of this study was to evaluate the influence of using the dental operating microscope (DOM) for detection of the mesiolingual (ML) canal orifice in extracted maxillary molars compared with unaided vision (no loupes or headlamps). STUDY DESIGN: Using a clinical simulation model system, we mounted 39 maxillary molars in a dentoform and placed them into a mannequin. After rubber dam placement and preparation of standard access, 2 attempts were made to locate the ML canal with unaided vision. Then the teeth were examined by using a DOM. Finally, all teeth were sectioned, stained, and evaluated with the DOM for actual presence of an ML canal. RESULTS: ML canal orifices were detected in 20 of the teeth with a sharp explorer and mirror. In the remaining teeth, 12 ML canal orifice were located by using the DOM. Qualitative nonparametric comparisons were used. CONCLUSIONS: The results of this study indicate that the DOM provides increased opportunity for the dentist to detect canal orifices. PMID- 11862211 TI - Color Doppler sonography of the facial artery in the anterior face. AB - OBJECTIVES: The purpose of this study was to clarify the Doppler sonographic features of the facial artery in the anterior face in healthy volunteers and to investigate those of related hemangiomas. STUDY DESIGN: Forty-six volunteers and 3 patients with hemangiomas were examined with Doppler sonography. The detection rates of the facial artery in the anterior face were determined. The features of the main trunk, superior and inferior labial branches, and the branches in the buccinator area were investigated together with the flow diameter, flow velocities, and resistive and pulsatility indexes. RESULTS: The detection rates of the main trunk and superior and inferior branches were 100%, whereas that of the branches in the buccinator area was 92.4%. There were no significant differences in the measured values for all indexes between the right and left sides. However, there were significant correlations between right and left sides in the flow diameter and minimum velocity of the main trunk and in the flow diameter and pulsatility index of the superior labial branch. The Doppler sonographic features of the hemangioma were characterized as a hypoechoic area with internal and surrounding blood flows. CONCLUSIONS: Doppler sonography can clearly depict the facial artery and its branches in the anterior face, and this method appears to be useful in the follow-up examination of hemangiomas in this area. PMID- 11862213 TI - Evaluation of digital and geometric unsharpness in dental radiographs using an endodontic file model. AB - OBJECTIVE: We sought to compare the effects of geometric and digital unsharpness. STUDY DESIGN: Endodontic files of sizes 6, 8, 10, and 15 were placed in the root canal of an extracted lower incisor in 3 different positions at 2-mm increments. Radiographs were taken with a GE 100 x-ray unit at magnification factors of 1.0, 1.1, 1.2, and 1.3. With the use of a flatbed scanner, each radiograph was digitized at resolutions of 75, 150, 300, and 600 dpi. Ten observers viewed the digital images and identified the positions of the end of the files and the end of the roots, as well as assessed image quality. RESULTS: No significant differences were found between the resolutions of 300 and 600 dpi, nor between the magnifications of 1.0 and 1.1. Significant differences were found between the magnification of 1.3 and the others and the resolution of 75 dpi and the others for all of the files. CONCLUSION: Geometric unsharpness has little relevance in clinical dental radiography. Increasing scanning resolution to 600 dpi did not improve accuracy with respect to recognizing the position of endodontic files. PMID- 11862212 TI - Whole-body (18)F-fluorodeoxyglucose positron emission tomography in patients with head and neck cancer. AB - OBJECTIVE: Because of the high risk of secondary primary malignancies in addition to possible distant metastases, whole-body evaluation is critically important in patients with head and neck cancer. We evaluated the clinical usefulness of whole body (18)F-fluorodeoxyglucose (FDG) positron emission tomography (PET) before initial treatment. STUDY DESIGN: We performed whole-body FDG PET in 26 patients with head and neck cancer (squamous cell carcinoma, n = 20; salivary gland carcinoma, n = 6) before initial treatment. FDG images were prospectively evaluated, and further imaging studies were performed if required. Final diagnosis for the presence or absence of distant lesions was made by analysis of the pathologic findings of surgical specimens or by analysis of the clinical follow-up data for more than 1 year. RESULTS: PET images showed FDG avid lesions distant from the head and neck area in 3 of 26 patients (11.5%). Two patients were confirmed to have secondary primary cancer (one with colon cancer in the early stage and another with small lung metastasis from postoperative colon cancer). They were diagnosed and treated properly both for the head and neck cancer and the secondary primary cancer. One patient was confirmed to have large lung metastasis from head and neck cancer, and appropriate treatment was selected. CONCLUSIONS: Whole-body FDG PET has a clinical impact on the management of patients with head and neck cancer by detecting secondary primary malignancies as well as distant metastases. PMID- 11862214 TI - A Drosophila APC tumour suppressor homologue functions in cellular adhesion. AB - Adenomatous polyposis coli (APC) is an important tumour suppressor in the intestinal epithelium. Its function in reducing nuclear beta-catenin and T-cell factor (TCF)-mediated transcription is conserved from Drosophila to mammals. But APC proteins are also associated with the plasma membrane. Here, we show that mutational inactivation of Drosophila E-APC causes delocalization of Armadillo (the Drosophila beta-catenin) but not DE-cadherin from adhesive plasma membranes. Extensive gaps between these membranes are visible at the ultrastructural level. The oocyte is also mislocalized in E-APC mutant egg chambers, a phenotype that results from a failure of cadherin-based adhesion. These results indicate that Drosophila APC functions in cellular adhesion; these results could have implications for colorectal adenoma formation and tumour progression in humans. PMID- 11862215 TI - Nuclear architecture and spatial positioning help establish transcriptional states of telomeres in yeast. AB - Recent experiments have shown that gene repression can be correlated with relocation of genes to heterochromatin-rich silent domains. Here, we investigate whether nuclear architecture and spatial positioning can contribute directly to the transcriptional activity of a genetic locus in Saccharomyces cerevisiae. By disassembling telomeric silent domains without altering the chromatin-mediated silencing machinery, we show that the transcriptional activity of silencer- reporter constructs depends on intranuclear position. This demonstrates that telomeric silent domains are actively involved in transcriptional silencing. Employing fluorescent in situ hybridization (FISH) in combination with genetic assays, we demonstrate that telomeres control the establishment of transcriptional states by reversible partitioning with the perinuclear silencing domains. Anchoring telomeres interferes with their ability to assume an active state, whereas disassembly of silencing domains prevents telomeres from assuming a repressed state. Our data support a model in which domains of enriched transcriptional regulators allow genes to determine transcriptional states by spatial positioning. PMID- 11862216 TI - Integrins regulate GTP-Rac localized effector interactions through dissociation of Rho-GDI. AB - The proper function of Rho GTPases requires precise spatial and temporal regulation of effector interactions. Integrin-mediated cell adhesion modulates the interaction of GTP-Rac with its effectors by controlling GTP-Rac membrane targeting. Here, we show that the translocation of GTP-Rac to membranes is independent of effector interactions, but instead requires the polybasic sequence near the carboxyl terminus. Cdc42 also requires integrin-mediated adhesion for translocation to membranes. A recently developed fluorescence resonance energy transfer (FRET)-based assay yields the surprising result that, despite its uniform distribution, the interaction of activated V12-Rac with a soluble, cytoplasmic effector domain is enhanced at specific regions near cell edges and is induced locally by integrin stimulation. This enhancement requires Rac membrane targeting. We show that Rho-GDI, which associates with cytoplasmic GTP Rac, blocks effector binding. Release of Rho-GDI after membrane translocation allows Rac to bind to effectors. Thus, Rho-GDI confers spatially restricted regulation of Rac-effector interactions. PMID- 11862217 TI - dS6K-regulated cell growth is dPKB/dPI(3)K-independent, but requires dPDK1. AB - Genetic studies in Drosophila melanogaster underscore the importance of the insulin-signalling pathway in controlling cell, organ and animal size. Effectors of this pathway include Chico (the insulin receptor substrate homologue), dPI(3)K, dPKB, dPTEN, and dS6K. Mutations in any of these components have a striking effect on cell size and number, with the exception of dS6K. Mutants in dS6K affect cell size but not cell number, seemingly consistent with arguments that dS6K is a distal effector in the signalling pathway, directly controlled by dTOR, a downstream effector of dPI(3)K and dPKB. Unexpectedly, recent studies showed that dS6K activity is unimpaired in chico-deficient larvae, suggesting that dS6K activation may be mediated through the dPI(3)K docking sites of the Drosophila insulin receptor. Here, we show genetically, pharmacologically and biochemically that dS6K resides on an insulin signalling pathway distinct from that of dPKB, and surprisingly also from that of dPI(3)K. More striking, despite dPKB-dPI(3)K-independence, dS6K activity is dependent on the Drosophila homologue of the phosphoinositide-dependent protein kinase 1, dPDK1, demonstrating that both dPDK1, as well as dTOR, mediated dS6K activation is phosphatidylinositide 3,4,5-trisphosphate (PIP3)-independent. PMID- 11862218 TI - Polyglycylation domain of beta-tubulin maintains axonemal architecture and affects cytokinesis in Tetrahymena. AB - Polyglycylation occurs through the post-translational addition of a polyglycine peptide to the gamma-carboxyl group of glutamic acids near the C terminus of alpha- and beta-tubulin, and has been found only in cells with axonemes, from protists to humans. In Tetrahymena thermophila, multiple sites of polyglycylation on alpha-tubulin are dispensable. By contrast, mutating similar sites on beta tubulin has site-specific effects, affecting cell motility and cytokinesis, or resulting in cell death. Here, we address the lethality of a polyglycylation deficiency in T. thermophila using heterokaryons. Cells with a lethal mutation in the polyglycylation domain of beta-tubulin assembled axonemes that lack the central pair, B-subfibres and the transitional zone of outer microtubules (MTs). Furthermore, an arrest in cytokinesis occurred, and was associated with incomplete severing of cortical MTs positioned near the cleavage furrow. Thus, tubulin polyglycylation is required for the maintenance of some stable microtubular organelles that are all known to be polyglycylated in vivo, but its effects on MTs appear to be organelle-specific. PMID- 11862221 TI - Rational therapeutics: dose titration and monotherapy in clinical practice. PMID- 11862220 TI - Structural basis of BLyS receptor recognition. AB - B lymphocyte stimulator (BLyS), a member of the tumor necrosis factor (TNF) superfamily, is a cytokine that induces B-cell proliferation and immunoglobulin secretion. We have determined the three-dimensional structure of BLyS to 2.0 A resolution and identified receptor recognition segments using limited proteolysis coupled with mass spectrometry. Similar to other structurally determined TNF-like ligands, the BLyS monomer is a beta-sandwich and oligomerizes to form a homotrimer. The receptor-binding region in BLyS is a deeper, more pronounced groove than in other cytokines. The conserved elements on the 'floor' of this groove allow for cytokine recognition of several structurally related receptors, whereas variations on the 'walls' and outer rims of the groove confer receptor specificity. PMID- 11862219 TI - Down-regulation of diabetogenic CD4+ T cells by a soluble dimeric peptide-MHC class II chimera. AB - Type 1 diabetes is an organ-specific autoimmune disease that is mediated by autoreactive T cells. We show here that administration of a soluble dimeric peptide-major histocompatibility complex (pMHC) class II chimera (DEF) to prediabetic double-transgenic mice prevents the onset of disease or, in animals that are already diabetic, restores normoglycemia. The antidiabetogenic effects of DEF rely on the induction of anergy in splenic autoreactive CD4+ T cells via alteration of early T cell receptor signaling and stimulation of interleukin 10 secreting T regulatory type 1 cells in the pancreas. Soluble dimeric pMHC class II may be useful in the development of immunospecific therapies for type 1 diabetes. PMID- 11862222 TI - Biodistribution and autoradiographic localization of I-125--labeled synthetic Peptide in aortic atherosclerosis in cholesterol-fed rabbits. AB - I-125 labeled SP4 is a synthetic oligopeptide derived from apolipoprotein B of low-density lipoprotein that has been shown to localized in atherosclerotic plaques in experimental animals. However, its biodistribution and mechanism of localization need to be further elucidated. Twenty-four cholesterol-fed (CF) and 20 normal (NL) New Zealand White rabbits were injected with I-125-SP4 and killed 15 to 30 min (6 NL; 6 CF) or 2 h (14 NL; 18 CF) later. We obtained aortic autoradiograms and activity concentrations (% injected dose/gm) in aortic segments and other tissues. The uptake of I-125-SP4 was higher in CF than in NL rabbits in all aortic segments (p < 0.05). I-125-SP4 was cleared rapidly in both CF and NL rabbits with 60 to 70% of the injected dose cleared from the blood by 1 h. No statistically significant differences in radiotracer biodistribution were observed between NL and CF rabbits although activity tended to be higher in the liver, gallbladder, and intestine in NL rabbits and in the kidney and spleen in CF rabbits. Silver grains were distributed mainly on foam cells of the fatty streaks in aortic microautoradiograms from two additional rabbits that had been injected with I-125-SP4. There were 23,518 plus minus 15,878 (SD) grains/mm(2) in fatty plaques but only 14,669 plus minus 11,035 grains/mm(2) in media muscle (p < 0.0001 [9 sections, 17 areas evaluated] in an atherosclerotic animal) in injected animals and 13,439 plus minus 5,565 grains/mm(2) in media muscle (two sections, four areas) in the normal control animals (NS versus media of atherosclerotic animal). I-125-SP4 specifically localizes in aortic atherosclerotic plaques in CF rabbits. There is no significant difference in tissue distribution between normal and CF rabbits except in the aorta. Preliminarily, it appears that the site of tracer uptake is on foam cells and this suggests the possibility of relative specificity for fatty plaque. PMID- 11862223 TI - Analysis of vancomycin in the hindlimb vascular bed of the rat. AB - Previously, studies have demonstrated that the effects of both a laboratory produced vancomycin and a clinically available vancomycin were mediated, in part, by activation of both H(1) and H(2) receptors; however, other mechanisms may play a role in the vascular changes associated with vancomycin, since neither H(1) and H(2) receptor blockade has completely abolished the vasodilator responses to vancomycin in any model system. To study the mechanisms of vancomycin interactions in the hindlimb vascular bed of the rat, responses of two types of vancomycin preparations were studied. Vancomycin prepared for either clinical or laboratory use produced an initial short-lived period of vasoconstriction followed by a prolonged period of vasodilation in the hindlimb vascular bed. Responses to both the vancomycins and histamine on systemic arterial vasodilation were significantly decreased after administration of both the H(1)-receptor antagonist diphenhydramine and the H(2)-receptor antagonist famotidine. Verapamil, an L-type calcium channel blocker, significantly reduced the vasopressor responses to clinical vancomycin but not the vasopressor responses to laboratory vancomycin. Enalaprilat, and angiotensin-converting enzyme blocker, significantly reduced the vasodilator responses but not the vasoconstrictor responses of clinical vancomycin and significantly reduced the vasoconstrictor responses but not the vasodilator responses to laboratory vancomycin. Meclofenamate, a cyclo-oxygenase inhibitor, and N(omega)-L-nitro-L-arginine methyl ester (L-NAME), a nitric oxide synthetase inhibitor, had no significant effect on the biphasic responses with either vancomycin preparations. Atropine, an anticholinergic-antimuscarinic receptor antagonist, and propranolol, a beta adrenergic blocker, had no significant effect on vancomycin responses. Finally, ondansetron, a serotonin receptor blocker, and HOE 140, a bradykinin receptor blocker, also had no significant effect on vancomycin responses. These data suggest that both vancomycin preparations (clinically available and laboratory prepared) caused biphasic responses that differed from the dose-dependent vasodilation elicited by histamine. Both vancomycin preparations' vasodilator responses appear to be modulated, in part, by a histamine receptor--sensitive mechanism, while vancomycin-induced vasoconstrictor responses appear to be modulated, in part, by angiotensin-converting enzyme and L-type calcium channel- sensitive mechanisms in the rat hindlimb vascular bed. These data also suggest that the vascular responses of vancomycin are preparation dependent. PMID- 11862224 TI - Pharmacokinetics and safety of a selegiline transdermal system relative to single dose oral administration in the elderly. AB - This open-label, two-phase cross-over study compared the safety and pharmacokinetics of transdermally administered selegiline and orally administered selegiline hydrochloride in elderly men and women (n = 6/gender). Single oral doses of 10 mg selegiline hydrochloride and single 1/2 and 1 selegiline transdermal system (STS) (delivering similar3.4 and 6.3 mg over 24 h) administered topically were safe and well tolerated in all subjects. Plasma concentrations of selegiline (SEL) and its N-desmethylselegiline (DMS), L amphetamine (AMP), and L-methamphetamine (MET) metabolites were measured using an HPLC/MS/MS method with lower quantitation limits of 10, 50, 200, and 200 pg/mL, respectively. No significant gender-related differences were observed following single 10-mg oral doses of selegiline hydrochloride or single 24-h applications of 1/2 and 1 STS to elderly males and females. The low level of dermal irritation as assessed by erythema and edema rating scales suggests that the STS was similar to Band-Aid (Johnson & Johnson, Skillman, NJ) controls. The transdermal administration of SEL bypasses the first-pass metabolism, that is significant after oral administration (first-pass extraction >90%). Peak plasma levels of 1.19, 23.22, 4.78, and 14.08 ng/mL were observed for SEL, DMS, AMP, and MET after a single 10-mg oral dose to the elderly. By contrast, peak plasma levels of 2.10, 0.85, 1.06, and 2.71 ng/mL were observed for SEL, DMS, AMP, and MET after a single 24-h application of 1 STS. Comparison of dose-corrected areas under the curve (AUCs) (made under the assumption of linear pharmacokinetics) indicate the SEL exposure after transdermal application was more than 50-fold greater than that obtained orally. This increase in systemic SEL exposure at the expense of metabolite formation that is reduced to <70% of that obtained orally for N-DMS, L AMP, and L-MET is hypothesized to be of therapeutic value in patients with a variety of neurodegenerative and psychiatric disorders. PMID- 11862225 TI - Evaluation of unbound serum carbamazepine and carbamazepine-10,11-epoxide concentration prediction methods in polytherapy adult patients with epilepsy. AB - We retrospectively evaluated the ability of equations with in vivo population binding parameters of our previous study (Method 1) or an average unbound fraction of 0.25 of Pynnonen (Method 2) to predict the unbound serum carbamazepine (CBZ) concentration in 35 serum samples from 18 adult patients with epilepsy receiving polytherapy. In 9 serum samples from 6 patients, the ability of equations for unbound serum carbamazepine-10,11-epoxide (CBZ-E) concentration prediction was also determined in predictive performance with in vivo population binding parameters of our previous study (Method A) or an average unbound fraction of 0.5 of Pynnonen (Method B). Mean prediction error, mean absolute prediction error (MAE), and root mean squared error (RMSE) were calculated for each method, and these values served as a measure of prediction bias and precision. Method 1 shows a bias to underpredict unbound serum CBZ. The MAE and RMSE were smaller in Method 2 (MAE = 0.454 &mgr;m/L, RMSE = 0.597 &mgr;m/L) than Method 1 (MAE = 0.597 &mgr;m/L, RMSE = 0.721 &mgr;m/L). Method 2 is superior to Method 1 in accuracy and precision. The effects of antiepileptic comedications on predictive performance of Methods 1 and 2 were determined in each group of serum samples with (n = 18, Group 1) or without (n = 17, Group 2) valproic acid (VPA) comedication. The results obtained by Method 1 show a bias to underprediction in Group 1 and no bias to over- or underprediction in Group 2. Results obtained by Method 2 show no bias to over- or underprediction in Groups 1 and 2. The effects of VPA comedication on predictive performance of unbound serum CBZ are relatively larger in Method 1 than Method 2. There was a weak but significant positive relationship between age and unbound serum CBZ fraction by simple regression analysis (n = 35, r = 0.368, p = 0.0297). The determined coefficient indicated that only about 14% of variations in unbound serum CBZ fraction can be explained by age, however. In each of Groups 1 and 2, no significant relationship was observed between age and unbound serum CBZ fraction. The effects of age on predictive performance of unbound serum CBZ are relatively small in patients receiving polytherapy. For unbound CBZ-E prediction, each of Methods A and B has no bias to over- or underprediction. The MAE was larger in Method B (MAE = 0.311 &mgr;m/L) than Method A (MAE = 0.233 &mgr;m/L). The differences in RMSE were small between Methods A (RMSE = 0.349 &mgr;m/L) and B (RMSE = 0.333 &mgr;m/L), however. Each of Methods A and B may have similar accuracy and precision. PMID- 11862226 TI - Monitoring of procainamide and N-acetylprocainamide concentration in saliva after oral administration of procainamide. AB - The aim of this study was to evaluate saliva as a potential monitoring medium for procainamide (PA) and its metabolite, N-acetylprocainamide (NAPA). Saliva concentrations of PA and NAPA were determined both in single and repeated oral administration of PA in four healthy subjects. PA and NAPA were detected both in serum and saliva after 500 mg of single oral administration of PA. After single oral administration, serum and saliva concentrations of PA and NAPA reached peak levels at about 1 h and declined thereafter. The mean half-lives of PA were 2.35 h in serum and 1.28 h in saliva. The mean half-lives of NAPA were 5.29 h in serum and 5.01 h in saliva. In this study, PA and NAPA concentrations in saliva were nearly twice as high as those in serum upon chronic oral administration as well as those in a single oral dose of PA. Significant correlation coefficients were observed between serum and saliva concentrations of PA (r = 0.78, p < 0.001, n = 21) and NAPA (r = 0.76, p > 0.001, n = 21) in single oral administration of PA. Significant correlation coefficients were also observed between serum and saliva concentrations of PA (r = 0.89, p < 0.001, n = 17) and NAPA (r = 0.87, p > 0.001, n = 19) after repeated oral administration of PA. The saliva-to-serum ratios of PA and NAPA maintained nearly constant at 1 h after oral administration. It would appear from this study that saliva is a suitable medium for monitoring PA and NAPA concentration regarding acetylator status. PMID- 11862227 TI - Postmenopausal osteoporosis: an overview. PMID- 11862228 TI - Postmenopausal osteoporosis. PMID- 11862229 TI - Envenomation from the brown recluse spider: review of mechanism and treatment options. AB - The brown recluse spider in commonly found throughout the midsouth region of the United States. Bites from the brown recluse occur when the spider is trapped in clothing or its nest is otherwise disturbed. The bite may be undetected by the patient until hours or days later when a characteristic lesion develops. Mild reactions to envenomation are usually limited to a lesion only. In some cases, a severe reaction results which can be life-threatening. Although there have been case reports of various pharmacological agents used for the treatment of brown recluse bites, none have been shown to be consistently effective. Therapy for brown recluse bites remains centered around aggressive wound care. Early surgical excision has not been shown to be of benefit and in most cases delays healing. This review focuses on the physiological mechanisms of the brown recluse venom and current treatment options. PMID- 11862230 TI - Gender differences in drug therapy. AB - Patient response to drug therapy is affected by multiple factors, including age, renal function, diet, smoking, congestive heart failure and gender. However, the data on gender differences in pharmacotherapy are limited, predominantly because women are excluded from clinical research due to the potential risks involved during pregnancy and co-existing illnesses in elderly women. Currently some information is available on gender differences in drug therapy. Differences in drug distribution and response due to different body composition as well as the effects of hormonal differences are the few that have been documented. Differences in drug metabolism due to differences in blood concentration of enzymes such as alcohol dehydrogenase has been recently described. Pharmacotherapy may also have to be altered due to differences in disease expression, like increased vasomotor tone in women. Drug efficacy has been found to be affected by gender---an example is antiplatelet therapy for stroke prevention. Finally, the side effects of drugs may differ depending on gender. Those involved in pharmacotherapy should be aware of these differences when prescribing drugs to female patients, since the majority of clinical data regarding pharmaceuticals has been derived primarily from a population of young males. PMID- 11862231 TI - The Therapeutics of Hypertensive Emergency. PMID- 11862232 TI - Long-Term Treatment with Oral Torsemide and Its Effect on Body Weight and Fluid Balance in Patients with Chronic Renal Insufficiency. AB - Long-term treatment with oral torsemide was studied to determine its effectiveness in maintaining steady-state fluid balance in patients with chronic renal insufficiency by using a placebo-controlled, double-blind, random-off design. Patients with stable chronic renal insufficiency were initially titrated and then stabilized on torsemide. Once stabilized on torsemide, patients were randomly assigned in a double-blind fashion to continue on their titrated dose of torsemide or to receive a placebo. Of the 82 patients enrolled in the study, 68 were randomized to torsemide (n = 34) or placebo (n = 34). Patients who received the placebo showed a significantly greater (p < 0.001) mean increase in body weight (3.55 lb) than did patients who remained on torsemide (0.46 lb). Approximately two-thirds of the weight gain observed in the placebo group occurred during the first 3 days after randomization. Patients continued to receive treatment unless they developed fluid accumulation that was considered deleterious to their clinical state as determined by the investigator. In the placebo group a greater number of patients discontinued treatment because of weight gain or fluid accumulation. The mean number of days on treatment after randomization was significantly higher (p < 0.001) for patients who received torsemide (26 days) than for patients who received the placebo (16 days). The lack of weight gain in the torsemide group was associated with a higher percentage of patients who showed no change or an improvement in peripheral edema status (79%) than in placebo patients (35%). No patient was withdrawn from the study because of hyperkalemia or hypokalemia. The adverse effects reported during the study were as anticipated for patients with chronic renal insufficiency that is often complicated by other underlying illnesses. PMID- 11862233 TI - Phase II Trial of Paclitaxel in Patients with Advanced Colon Cancer Previously Untreated with Cytotoxic Chemotherapy: An Eastern Cooperative Oncology Group Trial (PA286). AB - Paclitaxel was administered as a 24-h continuous infusion at 250 mg/m(2) in this Phase II trial in patients with adenocarcinoma of the colon or rectum. Nineteen patients were evaluated for toxity and 15 were assessable for response. There were no complete or partial responses, and toxicity present was primarily neutropenia. This study found that paclitaxel as a single agent does not have activity in adenocarcinoma of the colon or rectum. PMID- 11862234 TI - Effect of Pravastatin in the Treatment of Patients with Type III Hyperlipoproteinemia. AB - OBJECTIVE: To determine the efficacy and safety of pravastatin in the treatment of subjects with Type III hyperlipoproteinemia. DESIGN: Randomized, double-blind, placebo-controlled, crossover study. SETTING: Four lipid research clinics in the United States. PATIENTS: Twenty subjects between 18 and 70 years old with three diagnostic features of Type III hyperlipoproteinemia: very-low-density lipoprotein cholesterol (VLDL-C)/total plasma triglyceride ratio in excess of 0.30; beta-migrating VLDL pattern on agarose-gel electrophoresis; and the homozygous apolipoprotein E phenotype E(2)/E(2). After 4 weeks of dietary control, the subjects were eligible if their mean plasma total cholesterol level was at least 250 mg/dl and mean plasma trigylceraide level was at least 220 mg/dL. INTERVENTIONS: Subjects were randomly assigned to pravastatin 40 mg hs or placebo hs at the start of the first 6-week double-blind treatment period. After completing this phase, subjects entered a 4-week placebo/drug washout phase before crossing over to the opposite treatment for the second 6-week double-blind treatment period. MEASUREMENTS: Plasma VLDL-C and low density lipoprotein cholesterol (LDL-C) after ultracentrifugation and total triglyceride, cholesterol and high-density lipoprotein cholesterol (HDL-C) after 6 weeks of treatment; adverse clinical events and abnormal laboratory results. RESULTS: After 6 weeks of pravastatin theraphy, plasma levels of VLDL-C and LDL-C decreased 49% and 39%, respectively (p less-than-or-equal 0.01 vs. placebo). Total cholesterol and triglyceride concentrations decreased 36% and 22%, respectively (p less-than-or equal 0.001 vs. placebo). Levels of HDL-C increased 8% (p less-than-or-equal 0.01 vs. placebo). Pravastatin was tolerated well. One marked laboratory abnormality, an asymptomatic elevation of creatine phosphokinase, resolved upon completing pravastatin treatment. CONCLUSIONS: Pravastatin lowers plasma VLDL-C, LDL-C, total cholesterol, and triglyceride and raises HDL-C in subjects with Type III hyperlipoproteinemia. The safety profile is excellent. Pravastatin reductase inhibitor therapy affords a useful approach to the management of Type III or remnant removal disease. PMID- 11862235 TI - Pharmacokinetics of HFA-134a: A Preliminary Report. AB - HFA-134a blood levels from four clinical studies were combined to give a preliminary description of its population pharmacokinetics. HFA-134a was absorbed promptly and was eliminated with a half-life of 5.1 min. PMID- 11862236 TI - Approaches to Intravenous Nitroglycerin Therapy. PMID- 11862237 TI - Warfarin and Other "Anti"-Vitamin K Anticoagulants: Pharmacodynamics and Clinical Use. AB - Fortuitously discovered after an episode of bovine hemorrhagic disease, the oral antivitamin K anticoagulants inhibit a posttranslational modification of coagulation factors II, VII, IX, and X that is essential for bioactivity. They also influence formation of regulatory proteins C and S. Although their influence on the extrinsic pathway may be measured using the prothrombin time, a modification thereof produces the International Normalized Ratio (INR) which should allow for less interlaboratory variation. Finding utility in both prophylaxis and treatment of thromboembolic disease, these agents have rendered themselves important pharmacologic tools in the management of deep venous thrombosic, nonvalvular atrial fibrillation, structural heart disease, myocardial infarction, and following major surgical procedures. PMID- 11862239 TI - The Year in Review. PMID- 11862238 TI - Avoiding Overmedication of Elderly Patients. AB - Overmedication is the use of drugs that are not clinically indicated and are given in an inappropriate or overly aggressive manner even when nonpharmacologic alternatives are more suitable. This is an extension of polypharmacy and is caused when age-related changes in pharmacokinetics, pharmaco-dynamics, and potential drug interactions are ignored, when inappropriate drugs are used, or when a drug is prescribed for the wrong diagnosis in elderly individuals. Overmedication can cause adverse drug reactions, increased drug costs, and noncompliance with therapeutic regimens. This phenomenon can occur in the acute care or long-term care setting, and the community-dwelling elderly can also be exposed through the use of prescription and nonprescription drugs. Strategies to prevent overmedication in the elderly are discussed. PMID- 11862240 TI - Subclinical Hypothyroidism and Euthyroid Sick Syndrome in Patients with Moderate to-Severe Congestive Heart Failure. AB - Thyroid function tests were performed on baseline plasma that had been taken from 34 patients with NYHA Class II or Class III congestive heart failure (CHF). All patients were negative for thyroid disease on history and physical examination and none was taking medication known to alter thyroid metabolism. Analysis of thyroid function revealed abnormalities in 16 of 31 patients. These abnormalities fell into two categories: nine patients had elevated baseline thryroid stimulating hormone (TSH) above the normal limit while only one of these nine had subnormal thyroxine (T(4)) concentrations, suggesting the possibility of subclinical hypothyroidism. Seven patients demonstrated changes consistent with euthyroid sick syndrome (ESS). Weak correlations were observed between age and concentrations of T(4) and tri-iodothyronine (T(3)) and this suggests that changes in thyroid function cannot be explained solely on the basis of age. Although previous studies have demonstrated the presence of ESS in CHF, the present study suggests the possibility of a significant prevalence of subclinical hypothyroidism. PMID- 11862241 TI - Nitrate-Induced Headache in Patients with Stable Angina Pectoris: Beneficial Effect of Starting on a Low Dosage. AB - BACKGROUND: Nitrates, although important for the management of angina pectoris, cause significant headache in many patients. METHODS: In a randomized, double blind crossover study, 89 patients with stable angina pectoris were used to compare two different dosage strategies of isosorbide-5-mononitrate (5-ISMN). Patients were randomized to either 60 mg 5-ISMN once daily (o.d.) for 2 weeks or 30 mg 5-ISMN o.d. for 1 week followed by 60 mg 5-ISMN o.d. for 1 week. A 2-week placebo wash-out ensued, after which the alternative treatment was given. We assessed the occurrence of angina pectoris and headache by diary cards while taking into account the numbers of isosorbide dinitrate sublingual puffs and paracetamole tables required. Data were assessed for carryover and time effects. RESULTS: The two dosage regimens were equally efficient for the relief of angina pectoris without development of tolerance. Thirty percent of the patients never experienced headache from the given dosages. The remainder showed a highly significant time-effect: The total numbers of headache attacks in the 1st period of active treatment were 2,380 vs 1,400 attacks is the 2nd period (p < 0.003), yet significantly fewer patients had headaches on low dosages than high ones (45 vs 57, p < 0.02). CONCLUSIONS: Starting on a low dosage was associated with reduced frequency and severity of headache and did not notably influence the beneficial effect of nitrates on angina pectoris. One in three patients never experienced headache from the given dosages. The overall number of headache attacks in the 1st period of active treatment was significantly higher than that of the 2nd period, irrespective of the dosages given. PMID- 11862242 TI - Pharmacokinetics and Pharmacodynamics of a New Calcium Channel Blocker MPC-1304 in Hypertensive Subjects. AB - The pharmacokinetic and pharmacodynamic properties of a new calcium channel blocker MPC-1304 (MPC) were examined following a single 10 mg oral dose of MPC in six hypertensive subjects. Blood samples for measurement of MPC and its active metabolite (MPC-Keto-H(2)) were obtained, and blood pressure was measured just before and at 1, 2, 4, 6, 8, 12 and 24 h after administration. Plasma concentrations of MPC-Keto-H(2) were significantly higher than those of MPC at every observation point. The value of maximum plasma concentration of MPC-Keto H(2) was 10 times greater than that of MPC. The antihypertensive effect of MPC was observed at 4 h after administration and persisted up to 8 h. A plot of plasma concentrations of MPC versus reductions in blood pressure showed an anticlockwise hysteresis loop. These results suggest that the active metabolite MPC-Keto-H(2) contributes in part to the antihypertensive action of MPC, the delayed distribution of MPC into the effector site, or both. PMID- 11862243 TI - Safety and Efficacy of Once-Daily Captopril Plus Hydrochlorothiazide versus Nifedipine Gastrointestinal Therapeutic System in Black Patients with Mild to Moderate Hypertension. AB - The safety and efficacy of captopril plus hydrochlorothiazide (HCTZ) were compared to nifedipine gastrointestinal therapeutic system (GITS) in 145 randomly assigned black patients with mild to moderate hypertension. Following a 4-week placebo lead-in, patients received captopril plus HCTZ 25/15 mg or nifedipine GITS 30 mg for up to 12 weeks. Upward dose titration was permitted at weeks 3 and 6. Mean seated systolic and diastolic blood pressures decreased 16.1 +/- 13.5 mm Hg and 11.5 +/- 7.4 mm Hg, respectively, with captopril plus HCTZ. Statistically similar decreases were observed with nifedipine GITS: systolic, 19.3 +/- 12.2 mm Hg; diastolic, 13.8 +/- 7.2 mm Hg. There were no clinically significant between group differences in serum chemistries. Edema was reported in 20.3% of nifedipine GITS patients versus 1.4% of captopril plus HCTZ patients (p = 0.001). The two regimens were equally effective in controlling blood pressure in black patients; however, a higher incidence of edema occurred with nifedipine GITS compared to captopril plus HCTZ. PMID- 11862244 TI - Evaluation of an Auditory Feedback Equipped Metered Dose Inhaler. AB - BACKGROUND: Although metered dose inhalers are one of the most frequently prescribed drug delivery systems for pulmonary disease, patients usually do not employ ideal technique when using these systems. We studied the effect of a new teaching device, the MDI Tutor (MDI Tutor Inc., Palatine, IL), on patients with unsatisfactory inhalation technique. METHODS: Twelve patients with an average of 10 years' experience with metered dose inhalers (mean age, 41 years) were evaluated by two observers. All patients had deficits in their inspiratory technique such as actuation of inhaler coordinating with the beginning of inhalation cycle, uninterrupted inhalation, and incorrect length of inspiration. Following evaluation, the patients were given metered dose inhalers equipped with a MDI Tutor prototype as well as written instructions on how to use the device most effectively. RESULTS: All patients corrected their inspiratory technique deficits. CONCLUSIONS: Equipping a metered dose inhaler with a MDI Tutor allows patients to rapidly recognize correct the errors in their inspiratory technique. PMID- 11862245 TI - A Review of Post--Cardiopulmonary Bypass Bleeding, Aminocaproic Acid, Tranexamic Acid, and Aprotinin. AB - OBJECTIVE: To review the mechanism and cause of hemostatic defects following cardiopulmonary bypass (CPB) and determine the safety and efficacy of antifibrinolytic agents for use in cardiac surgery and patients likely to benefit. DATA SOURCES: A MEDLINE search (1966 to present) of the English-language literature pertaining to aminocaproic acid (ACA), tranexamic acid (TA), and aprotinin was performed. Additional literature was obtained from reference citations of pertinent articles identified through the search. STUDY SELECTION AND DATA EXTRACTION: While all articles of relevance were considered for inclusion, this review evaluates only clinical trials with emphasis on prospective, randomized, controlled studies. DATA SYNTHESIS: In reported trials, ACA and TA each reduce mediastinal blood losses by about one-third, while transfusion needs remain unchanged. ACA and TA dosing inconsistencies, omission of transfusion criteria, and unidentified surgical risk factors prevent optimal findings. Thromboembolic complications could not be ascribed to either ACA or TA in more than 950 patients studied. Aprotinin decreased mean mediastinal blood losses by 42%, 67% and 48% in primary coronary artery bypass grafting (CABG), reoperative CABG, and in CABG patients receiving aspirin, respectively. Transfusion needs were reduced 42% in primary CABG patients and 55% to 88% in high-risk patients. Patients at high risk of bleeding (i.e., reoperative CABG and patients on aspirin) demonstrated greater transfusion needs and blood loss than primary CABG patients. As blood conservation measures may eliminate the need for transfusions among primary CABG patients, patients at higher risk may benefit most from the addition of antifibrinolytic prophylaxis. CONCLUSION: The efficacy of all antifibrinolytic agents in cardiac surgery has been established, but comparative data is inconclusive to suggest an agent of choice. Thromboembolic complications have been rare and difficult to ascribe to antifibrinolytic agents. Future trials comparing efficacy of agents in high risk patients and rigorously evaluating thromboembolic events will allow unconditional recommendations. PMID- 11862246 TI - Therapeutic Drug Monitoring in the Elderly. AB - Therapeutic drug monitoring (TDM) is defined as the strategy by which the dosing regimen for a patient is guided by repealed measurements of plasma drug concentrations. TDM is also used to confirm drug poisoning and toxicity, to diagnose adverse drug reactions, and to verify patient compliance with drug regimens. There is a scarcity of data about the use of TDM in treating the elderly, but general inferences can be made based on pharmacokinetic and pharmacodynamic studies in older populations and on investigations in younger individuals. Despite its important limitations TDM seems to be an important tool in drug therapy management in the elderly given the increased frequency of chronic medical conditions, polypharmacy, drug interactions, and the atypical presentation of adverse drug reactions in this population. PMID- 11862247 TI - Nonphysician Prescribing. PMID- 11862248 TI - The Effect of Misoprostol and Prostanoids On cAMP Production and Calcification in a Differentiating Chick Limb-Bud Culture System. AB - Mesenchymal cells isolated from chick limb-buds, when plated in micromass culture, differentiate into chondrocytes, forming a mineralizable matrix. Because these cells produce prostanoids during differentiation, this system was used to test the hypothesis that E-series prostanoids are involved in chondrocyte mediated calcification. Prostaglandins E(2) (PGE(2)) and E(1) (PGE(1)), and the E series analog, misoprostol (MP), increased chondrocyte cAMP content in the presence of the phosphodiesterase inhibitor IBMX. The increases for PGE(1) and PGE(2) at their saturation concentrations were twofold to threefold greater than for MP at its saturation concentration. At culture day 7, 9, or 11 (the day that mineralization commenced), the maximal cyclic adenosine monophosphate (cAMP) production was at a concentration of 250--500 ng/ml PGE(2), 500--1000 ng/ml PGE(1), and less-than-or-equal5000 ng/ml MP. At these concentrations, PGE(1) and PGE(2), but not MP, stimulated chondrocyte differentiation. (45)Ca accumulation in mineralizing, as compared to nonmineralizing, similarly treated control cultures was not altered by the addition of indomethacin and/or prostanoid when the phosphate source was inorganic phosphate. Because the prostanoids decrease alkaline phosphatase activity, initial beta-glycerophosphate-mediated mineralization was inhibited by each of the prostanoids. PMID- 11862249 TI - Nonsteroidal Anti-Inflammatory Drugs and Prostaglandins: Their Interactions and Effects on the Particulate-Induced Inflammatory Process Implicated in Joint Implant-Loosening and on Monosodium Urate Crystal-Induced Inflammation. AB - The objective of this study was to determine the effects of orally administered misoprostol and diclofenac on inflammation caused by particulates in the rat subcutaneous air-pouch model. Subcutaneous air pouches were formed in male Sprague-Dawley rats. The animals were premedicated by gavage with either saline, diclofenac, misoprostol, or both diclofenac and misoprostol. The air pouches were injected with either polymethyl methacrylate particles or monosodium urate crystals, and the pouch fluids were obtained at 1, 6, 24, 48, and 72 h. Leukocyte influx, tumor necrosis factor, neutral metalloprotease activity, and prostaglandin E(2) levels were measured. It was determined that leukocyte influx was inhibited by diclofenac in the acute inflammation caused by monosodium urate crystals only. In all animals receiving diclofenac, prostaglandin E(2) (PGE(2)) levels were reduced, whereas tumor necrosis factor levels were elevated. The elevation of tumor necrosis factor was prevented by the addition of misoprostol. It is concluded that the oral administration of diclofenac or misoprostol can affect levels of specific mediators involved in particulate-induced inflammation in the subcutaneous air pouch. PMID- 11862250 TI - Co-Stimulatory Effects of Misoprostol and Diclofenac on Glycosaminoglycan Synthesis in Human Cartilage Explants and Their Therapeutic Relevance. AB - Misoprostol, a prostaglandin E(1) analog, is currently available to manage ulcer disease, being used predominantly in the prophylaxis of nonsteroidal anti inflammatory drug (NSAID)-induced ulceration, a serious side effect of anti inflammatory therapy in arthritis. The protective effects of misoprostol have now also been shown to extend to cartilage in a series of experiment using an ex vivo system employing normal human and osteoarthritis (OA) cartilage. Misoprostol reproducibly reverses inhibition of proteoglycan synthesis induced by interleukin 1 and certain NSAIDs, and also stimulates synthesis in OA cartilage. The article reviews these findings and also presents the results obtained in a study of 20 OA cartilages in which synergy was demonstrated between misoprostol (at 50 ng/ml) and diclofenac (0.3 &mgr;g/ml) in preventing proteoglycan synthesis. Diclofenac on its own is unusual amongst NSAIDs in exerting virtually no deleterious effect on cartilage. The synergy with misoprostol is of clinical interest in view of the recent introduction of a misoprostol/diclofenac combination product (Arthrotec), the intention of which was to provide antiinflammatory efficacy with a reduced incidence of GI damage. The implications of these cartilage experiments is that such a combination may also offer improvements in the management of the arthritic process in OA, and methods whereby this would be assessed clinically are discussed. PMID- 11862251 TI - Experimentally Induced Articular Cartilage Injury: The Combined Use of Misoprostol and Diclofenac as Therapeutic Agents. AB - Injection of chymopapain (CP) into the rabbit knee causes rapid depletion of proteoglycans (PGs) in the articular cartilage. Successful repair of the articular cartilage follows injection of only 0.2 mg CP whereas in animals injected with 0.2 mg CP, the repair phase fails leading to cartilage destruction within 21 days. Previous work by the authors indicated that daily oral diclofenac reduced the severity and incidence of CP-induced cartilage injury after 21 days. The present study was designed to determine if administration of diclofenac promoted articular cartilage repair after CP injury over a longer period of time and if coadministration with misoprostol could reduce the concentration of diclofenac needed to promote repair as well as confer protection to the gastric mucosa. Adolescent male New Zealand white rabbits received CP-induced cartilage injury (2.0 mg) followed by daily oral diclofenac, diclofenac/misoprostol or misoprostol. Control animals received diclofenac, diclofenac/misoprostol, or misoprostol or no drug treatment with no cartilage injury. All animals were sacrificed after 2 months. Examination of the stomachs from animals receiving no drug or misoprostol only were completely normal. Most animals receiving diclofenac only had abnormal gastric mucosa. In contrast, coadministration of diclofenac/misoprostol revealed significantly fewer abnormalities. Analysis of the composite histologic score of patellae and measurements of cartilage PG content revealed no significant differences in groups which received CP injury with and without drug treatment. Histology scores and cartilage PG content from animals in groups which did not receive CP injury, but were treated with diclofenac/misoprostol, diclofenac or misoprostol were within normal limits and thus did not differ significantly from untreated control cartilage. The results of this study indicate that daily administration of diclofenac, diclofenac/misoprostol or misoprostol has no protective effect on CP-induced cartilage injury after 2 months. Administration of diclofenac only results in severe gastric abnormalities which are significantly reduced if diclofenac is coadministered with misoprostol. Administration of diclofenac, diclofenac/misoprostol or misoprostol has no effect on PG content of cartilage from otherwise normal joints. PMID- 11862252 TI - Comparative Effects of Ceftazidime Two or Three Times Daily in Patients with Skin and Skin Structure Infections. AB - BACKGROUND: Skin and skin structure infections are among the most common infectious diagnoses in both the hospital and community settings. If untreated, these infections can produce serious complications. Successful antimicrobial therapy for these infections requires coverage against the causative pathogens, particularly Staphylococcus aureus. OBJECTIVE: This randomized, single-blind, multicenter study was designed to compare the efficacy and safety of intravenous ceftazidime 2 g given two times daily (BID) with that of intravenous ceftazidime 1 g given three times daily (TID) for the treatment of skin and skin structure infections caused by ceftazidime-sensitive pathogens. METHODS: Adults (greater than-or-equal18 years) were eligible for enrollment if they were hospitalized or in home health care settings and they had a skin or skin structure infection caused by a ceftazidime-sensitive pathogen. Patients were randomly assigned to receive ceftazidime 2 g every 12 h or ceftazidime 1 g every 8 h as an intermittent infusion over 15--30 min. Treatment was continued for 2--3 days beyond the time the patient became asymptomatic or evidence of bacterial eradication was obtained; however, total treatment duration had to be at least 5 days. Patients were assessed for their clinical and bacteriological response at the end of treatment and for their clinical response at follow-up. RESULTS: A total of 806 patients were enrolled in the study, 406 of whom received ceftazidime 2 g BID and 400 of whom received ceftazidime 1 g TID. Both treatments were administered for a mean duration of 9 days. At the end of therapy, 248 of 264 (94%) clinically evaluable patients receiving ceftazidime BID and 258 of 275% (94%) clinically evaluable patients receiving ceftazidime TID achieved clinical cure or improvement (p = 0.953). Pathogens were eradicated or presumed to be eradicated from 217 of 256 (85%) bacteriologically evaluable patients receiving ceftazidime BID and from 228 of 266 (86%) bacteriologically evaluable patients receiving ceftazidime TID (p = 0.760). Of 1131 isolates, the most common pathogens were S. aureus (30%) and Pseudomonas aeruginosa (10%). Both regimens were well tolerated with only 13 patients (3%) in the BID group and 16 patients (4%) in the TID group withdrawing because of an adverse event. CONCLUSIONS: These data indicate that ceftazidime 2 g given twice daily is as effective as ceftazidime 1 g given three times daily for the treatment of skin and skin structure infections. In addition, the twice-daily regimen has the advantage of convenience. PMID- 11862253 TI - The Effect of Urinary pH and Flow Rate on the Renal Elimination of Zopolrestat and Zopolrestat Glucuronide in Humans. AB - Zopolrestat is an aldose reductase inhibitor that may be useful in the treatment of diabetic complications by reducing flux through the polyol pathway. The plasma half-life of zopolrestat in man is approximately 30 h, and approximately 45% of an orally administered 1000-mg dose is eliminated in the urine as unchanged drug. Because active secretion accounts for much of the renal clearance for zopolrestat, a carboxylic acid with a pK(a) of 5.46, the effect of urinary pH and flow rate on renal clearance of drug was investigated in a series of studies. Renal clearance of zopolrestat following oral administration of 200 mg was determined in normal male volunteers under basal conditions and after treatment with NH(4)Cl and NaHCO(3) to alter urinary pH. Plasma concentrations of zopolrestat were similar under basal and NaHCO(3) treatment but were approximately twofold higher under NH(4)Cl treatment. However, the half-life of zopolrestat under NH(4)Cl treatment (29.5 h) was similar to the half-life of zopolrestat in untreated subjects. Renal clearance decreased by a factor of 2.54 for each unit decrease in urinary pH. In a second study, there was no effect of urine flow rate on renal clearance following an oral dose of 400 mg. Renal elimination of zopolrestat and zopolrestat glucuronide was also examined in volunteers with normal urine flow dosed at either 600 or 1000 mg/day. Whereas renal clearance of zopolrestat decreased with decreasing urinary pH, renal elimination of zopolrestat glucuronide was not affected by pH. PMID- 11862254 TI - Is the Degree of Arrhythmia Suppression a Predictor of Survival? AB - The effectiveness of an antiarrhythmic drug is judged by the degree of ventricular arrhythmia (VA) suppression. We evaluated the relationship between the degree of VA suppression and survival in a dose-adjusted trial of 110 symptomatic patients treated with amiodarone. Cohorts had left-ventricular ejection fraction (LVEF) of 41 plus minus 18%, ventricular premature contractions (VPCs) of 445 plus minus 571 h, couplets (C) of 733 plus minus 1498 24 h and nonsustained (N) ventricular tachycardia (VT) of 65 plus minus 217 24 h; these conditions were followed for 15 plus minus 11.5 months. Amiodarone was initiated with an oral loading of 670 plus minus 111.7 mg per day for 10 days and continued on maintenance of 274.9 plus minus 102 mg per day. Survival rates of responders and nonresponders with VPCs <70%, 70--89%, greater-than-or-equal90%; C greater than-or-equal 90%; NVT (100%); and the response to all 3 criteria (suppresion of VPCs greater-than-or-equal70%, C greater-than-or-equal 90% and complete abolition of NVT) were not statistically significant. Survival rates as a function of LVEF <40% (51 patients) or greater-than-or-equal40% (59 patients), as well as responders or nonresponders to all three criteria, were not significant (p = NS). We conclude that, in patients treated with low-dose amiodarone, the degree of VA suppression of PVCs, C and NVT does not predict survival; the survival of patients with LVEF <40% improved irrespective of VA suppression; and criteria for VA suppression should be reassessed at lower levels of suppression for the improvement of the drug risk:benefit ratio. More improvement is not necessarily better. PMID- 11862256 TI - Neurohumoral Mechanisms in Congestive Heart Failure and the Role of Drugs with Multiple Actions: A Review of Carvedilol. AB - Treatments based on mechanistic models of heart failure, although entirely rational, have not succeeded in reducing mortality. A more holistic view, based on the concept that the neurohumoral environment is a common factor, encompasses all the previous theoretical models and extends them to provide a new approach to treatment. Vasodilators, inotropes, and angiotensin-converting enzyme (ACE) inhibitors all modify hemodynamics favorably and improve cardiac-performance. Only the vasodilators and ACE inhibitors have been shown to reduce morbidity and mortality. With the ACE inhibitors, this effect is related to changes in the levels of circulating angiotensin II and noradrenaline rather than to improved hemodynamics. There is direct evidence that adrenergic receptor density is reduced in chronic heart failure. Vasodilating beta-blockers, like carvedilol, that suppress adrenergic overactivity and reduce preload and afterload, offer a new approach to treatment, but the exploitation of this new opportunity is inhibited by the widely held belief that beta-blockers should be avoided in heart failure. This mistaken view is based on the experience of beta-blockers in angina pectoris and after myocardial infarction, experience that should not exclude their use in patients with congestive heart failure. Vasodilating beta-blockers, introduced at an appropriate dosage, may have an important role to play in the overall management of congestive failure. PMID- 11862255 TI - Combination Treatment with Sustained-Release Verapamil and Indapamide in the Treatment of Mild-to-Moderate Hypertension. AB - A multicenter study of the treatment of mild and moderate hypertension compared three once-daily regimens for efficacy and safety: Group I, verapamil sustained release (SR) 240 mg; Group II, verapamil SR 480 mg; and Group III, verapamil SR 240 mg plus indapamide 2.5. mg. After a 3-week placebo washout period and a 2 week preliminary treatment period with verapamil SR 240 mg, patients with sitting diastolic blood pressures of 95--115 mm Hg were randomized in a double-blind fashion to one of the three treatment groups for a duration of 16 weeks. (Patients with diastolic BP < 95 mm Hg on this initial treatment were excluded from further study.) Efficacy assessed after 16 weeks or at the end point showed all three treatments significantly reduced sitting diastolic blood pressure from baseline levels. Compared with Group I (low-dose) verapamil SR treatment, there was a reduction in Group III combination therapy of 8.6 mm Hg systolic and 3.0 mm Hg diastolic and a significant reduction in Group II high-dose verapamil SR therapy of 7.6 mm Hg systolic and 3.9 mm Hg diastolic BP. Efficacy results were independent of age or body weight. Patients who were poor responders to lead-in verapamil SR 240 mg daily prior to randomization tended to have a greater response to combination therapy than to high-dose verapamil SR. Adverse experiences leading to withdrawal from the study occurred in 21% of patients receiving high-dose verapamil (Group II), exceeding the withdrawal from the other two treatment groups (8.5% from Group II and 4% from Group I). The most frequent adverse experiences in Group II were constipation and peripheral edema; headache was most common in Groups I and III. This trial demonstrates that adding indapamide to low-dose verapamil SR enhances efficacy and is well tolerated. PMID- 11862257 TI - Aminoglycoside-Induced Nephrotoxicity in Cystic Fibrosis: A Case Presentation and Review of the Literature. AB - Nephrotoxicity manifested by renal insufficiency is a well-known consequence of aminoglycoside administration in hospitalized patients. We report a case of aminoglycoside-induced nephrotoxicity in a young woman with cystic fibrosis that illustrates the classical presentation of nonoliguric renal failure and the risk factors that predispose to the development of aminoglycoside renal toxicity. The incidence of aminoglycoside nephrotoxicity in cystic fibrosis appears to be lower than that expected for a population exposed to prolonged and repetitive courses of therapy with these agents. Cystic fibrosis is a unique disease in which the pharmacokinetic profile of many drugs is abnormal. The pharmacokinetics of aminoglycosides in cystic fibrosis are characterized by an increased drug clearance and a larger volume of distribution; why cystic fibrosis patients should exhibit these abnormalities remains an enigma. If, in fact, patients with cystic fibrosis have a resistant phenotype against aminoglycoside-induced renal damage; further research designed to investigate the complexity of the renal handling of these drugs in these subjects would contribute to unravel the molecular mechanisms of aminoglycoside nephrotoxicity. PMID- 11862258 TI - The Cancer Myth. PMID- 11862259 TI - The Analgesic Interaction of Misoprostol with Nonsteroidal Anti-Inflammatory Drugs. AB - The purpose of this project was to evaluate the analgesic efficacy of misoprostol when combined with ibuprofen or diclofenac Na. Animal experiments using the inflamed rat paw formalin model suggested that misoprostol potentiates the analgesic effect of some NSAIDs (nonsteroidal anti-inflammatory drugs) including diclofenac Na but not propionic acid derivatives or opiates. The dental pain model was used to evaluate the clinical relevance of this interaction. Patients received a single oral dose of study medication following surgical removal of impacted teeth. Patients were medicated for moderate to severe postsurgical pain and then filled in an analgesic diary for a 6-h observation period. Several blood samples were taken over the observation period. In addition, microdialysis samples were taken directly from the extraction socket and were analyzed for immunoreactive prostaglandin E(2) levels. The studies were single-dose, parallel group and double-blind assays. In the first study, 70 patients received an oral dose of either placebo (n = 13), misoprostol 200 &mgr;g (n = 18), ibuprofen 200 mg (n = 19), or the combination of misoprostol + ibuprofen (n = 20). Misoprostol alone demonstrated a small analgesic effect compared to placebo. Both the ibuprofen and combination groups were substantially more effective than placebo but not different from each other. The combination group had higher ibuprofen blood levels during the first 45 min but had a lower C(max) and longer time to T(max). The second study evaluated oral doses of placebo (n = 11), misoprostol 200 &mgr;g (n = 21), diclofenac Na 50 mg (n = 18), and the combination of misoprostol + diclofenac Na (n = 20). Relative to placebo, misoprostol performance was similar to the first study. When the results of the two studies were combined, there was a small, but statistically significant, analgesic effect for misoprostol. Diclofenac Na was superior to both placebo and to misoprostol alone. The combination was the most effective treatment, and for hours 4--6 it was significantly better than diclofenac Na alone. Analysis of the blood samples showed an earlier and higher peak effect for the diclofenac Na group compared to the combination, and the combination again had a lower C(max). The microdialysis probe assays demonstrated that misoprostol depressed PGE(2) levels at the peripheral site of trauma over the first 2 h after surgery. These pilot studies used small samples, and the results only suggest trend effects. Both studies demonstrated that misoprostol 200 &mgr;g, a prostaglandin analog, does have an analgesic effect. When combined with ibuprofen, there was no potentiation of analgesia. In contrast, the combination of misoprostol + diclofenac Na demonstrated an enhanced peak effect, total effect for pain intensity difference and pain relief (sum pain intensity difference [SPID] and total pain relief [TOTPAR]), and PMID- 11862260 TI - Evoked Release of Amino Acids and Prostanoids in Spinal Cords of Anesthetized Rats: Changes During Peripheral Inflammation and Hyperalgesia. AB - C-fiber stimulation of the sciatic nerve in rats with an acute experimental arthritis elicited a greatly enhanced spinal release of several amino acids and prostaglandin PGE(2) (PGE(2)) when compared to control rats. The most dramatic change was in aspartate (Asp) which was not released in the control state but did increase significantly for a period outlasting the stimulation in the animals with knee joint inflammation. Evoked release of Glu, Gly, and PGE(2) also increased in the experimental group. It is proposed that the Asp acts primarily at N-methyl-D-asparate (NMDA) receptors to participate in the generation of hyperalgesia. In a similar model, direct activation of the NMDA receptors evoked spinal release of the same amino acids, including citrulline (an index of nitric oxide production) as well as representative prostanoids. Pretreatment with a NO synthase inhibitor blocked the release of Cit and Glu and reduced PGE(2) release evoked by NMDA, indicating that 1) Glu release is downstream of the actions of NO, and 2) the cyclooxygenase system is not independent of NO production. Evoked release of thromboxane B(2) was not affected by pretreatment with the nitric oxide synthesis inhibitor. Pretreatment with a cyclooxygenase inhibitor antagonized release of both prostanoids, but had little effect on NMDA-evoked increases in amino acids and Cit. PMID- 11862261 TI - Modulation of Substance P Release in Primary Sensory Neurons by Misoprostol and Prostaglandins. AB - Pain is one of the cardinal signs of inflammation. A number of inflammatory mediators have been shown in animal models to induce or augment pain. Of particular interest are the prostaglandins (PGs), which are arachidonic acid metabolites and can be pharmacologically regulated by cyclooxygenase inhibitors, the nonsteroidal anti-inflammatory drugs (NSAIDs). Indeed, NSAIDs are potent modulators of pain sensitivity. They are thought to mediate their hypoalgesic action through inhibition of prostaglandin production. However, indiscriminate inhibition of prostaglandin synthesis also creates a significant number of clinical side effects, among them gastrointestinal toxicity. With the introduction of misoprostol, a PGE(1) analog, a large number of investigative possibilities are opened. We have proposed to study the effect of misoprostol in an in vitro pain model. Our model, created by culturing primary sensory neurons isolated from dorsal root ganglions, was then differentiated by nerve growth factor and subjected to electrical stimulation. Substance P release following electrical stimulation was quantitated by radioimmunoassay. We found that misoprostol augmented substance P release in a dose-related manner. With 100 ng/ml of misoprostol added, there was a 45% increase in substance P release as compared to control. PGE(1) and PGE(2) addition at similar concentration caused a similar degree of increase in substance P release. Thus, acute addition of misoprostol to cultured sensory neurons appears to sensitize them to release more substance P. Our result does not necessarily imply that misoprostol will cause pain clinically. In our study, misoprostol at 10 ng/ml has no effect on substance P release. Because the plasma concentration of misoprostol is in the picogram per milliliter range, misoprostol most likely does not have a pain potentiation effect at the recommended therapeutic dose. Our data indicated that misoprostol at therapeutic dose has an insignificant effect on substance P release from primary sensory neuron. PMID- 11862262 TI - Tools for Investigating Functional Interactions Between Lipid-Derived Autacoids and their Receptors. AB - A method for rapidly evaluating functional interactions between ligands and G protein--coupled receptors has been developed. The technology is based on the ability of animals to change color by controlling the position of pigmented organelles within skin cells called melanophores. cDNA coding for a receptor to be studied is expressed in immortalized frog melanophores. Stimulation of a receptor that normally functions to activate either adenyl cyclase or phospholipase C induces centrifugal melanosome translocation and cell darkening. Conversely, application of an agonist to cells expressing a receptor that operates to inhibit adenyl cyclase induces centripetal pigment movement and cell lightening. The simple optical change can be used to investigate ligand-receptor interactions at several levels, including single-cell analysis and high throughput chemical screening. Current efforts are focused on (1) identifying small peptides that activate or block thromboxane. A(2) and platelet-activating factor (PAF) receptors and (2) cloning eicosanoid receptors. PMID- 11862263 TI - Phospholipase A(2) and Lipids as Potential Modulators of c-Raf-1 Kinase. AB - c-Raf-1 is a proximal serine/threonine kinase in the signaling cascade of many mitogens. The cellular mechanisms responsible for regulation of this kinase remain ill-defined. Although c-Raf-1-associated proteins have been identified, including Ras, none of these have been found to activate c-Raf-1 kinase in vitro. To evaluate whether arachidonic acid or one of its products is implicated in c Raf-1 activation, c-Raf-1 activity was measured in LLC-PK(1) kidney epithelial cells overexpressing the 100 kDa phospholipase A(2) (PLA(2)). As compared to control neomycin plasmid transfected cells, the cells overexpressing PLA(2) had a greater activation of c-Raf-1 in response to A23187 and phorbol ester stimulation. To explore the possibility that c-Raf-1 activity may be modulated directly by lipids, the enzymatic characteristics of c-Raf-1 were determined, and the effects of various possible lipid modulators on c-Raf-1 activity were examined. The K(m) of c-Raf-1 for ATP and mitogen-activiated protein kinase kinase (MAPKK), the only known physiologic substrate of c-Raf-1, were 11.6 &mgr;M and 0.8 &mgr;M, respectively. Of 13 lipids or combinations of lipids tested, including arachidonic acid and several eicosanoids, only phosphatidylserine and diacylglycerol in the presence of CA(2+) (2.5 mM) increased c-Raf-1 kinase activity significantly. The increase (1.5-fold) was approximately two orders of magnitude less than the stimulation of protein kinase C by these lipids. c-Raf-1 kinase activity and immunoreactivity eluted on gel filtration at a predicted molecular mass of greater than 150 kDa, suggesting that active c-Raf-1 is part of a multimeric complex. The absence of immunoreactive Ras in the active fractions confirms that the interaction is not necessary to maintain c-Raf-1 in an active state. In conclusion, a product of PLA(2) may play a role, together with Ras and another unidentified cofactor, in activating c-Raf-1. This lipid mediator(s) may directly or indirectly regulate the activity of c-Raf-1, but the identity of the mediator and its mode of interaction with c-Raf-1 and its associated proteins remain unclear. PMID- 11862264 TI - Effect of Felbamate on The Pharmacokinetics of Clonazepam. AB - To assess the possible interaction between clonazepam and felbamate, a double blind, randomized, placebo-controlled, two-way crossover study was conducted in 18 healthy male volunteers. Volunteers were administered clonazepam (1 mg q12h) and felbamate (1200 mg q12h) or matching placebo for 10 days during each period of the crossover. Following morning dosing on day 10, blood samples were obtained over 12 h for the determination of clonazepam and the metabolites 7-amino clonazepam and 7-acetamido-clonazepam. Felbamate increased clonazepam's C(max) and AUC(0--12 h) by 17% and 14%, respectively (p < 0.01). The 90% confidence intervals following log-transformation for each of these pharmacokinetic parameters were within the generally accepted interval (80--125%) for bioequivalence. Felbamate had no significant effect on the pharmacokinetics of 7 amino-clonazepam, whereas 7-acetamido-clonazepam concentrations were below the limit of quantification in all but one subject. Adverse events were mainly central nervous system in nature, with a greater incidence reported during coadministration with felbamate compared with placebo. Overall, felbamate appears to have no clinically relevant effects on the pharmacokinetics of clonazepam. PMID- 11862265 TI - The Effect of Dosing Regimen and Food on the Bioavailability of the Extensively Metabolized, Highly Variable Drug Eldepryl((r)) (Selegiline Hydrochloride). AB - The pharmacokinetics of oral selegiline hydrochloride were examined in two crossover studies in healthy volunteers: a 5-mg b.i.d. administration in the fed versus-fasted state and a 5-mg b.i.d. versus 10-mg q.d. multiple-dose administration. Food increased the systemic exposure of selegiline, while metabolite levels (N-desmethylselegiline, L-amphetamine and L-methamphetamine) were unaffected. Mean selegiline C(max) and AUC(t) increased by threefold in the fed state. Following multiple doses, selegiline and metabolites accumulate from single-dose exposure. The extent of accumulation was greatest for selegiline. Trough levels of 3.3 and 6.0 ng / mL for L-amphetamine and L-methamphetamine suggest that steady state was achieved by the third dosing day for these metabolites. The mean half-life of selegiline and N-desmethylselegiline of 8.6 and 9.5 h, respectively, was substantially longer than that obtained in single dose experiments (1.5 and 3.8 h, respectively). Once- and twice-daily regimens were not equivalent with respect to selegiline pharmacokinetics. While minor differences in N-desmethylselegiline pharmacokinetics was observed, L-amphetamine and L-methamphetamine pharmacokinetics were unaffected by dosing regimen. Pharmacokinetic parameters describing selegiline plasma levels from a single 10 mg dose did not predict the plasma concentrations obtained from 5-mg b.i.d. dosing. The effects of food and regimen on selegiline pharmacokinetics are consistent with highly extracted compounds in which absorption or metabolism is site specific. These factors contribute to the high variability in selegiline plasma levels following oral dosing. The prolonged half-lives of selegiline and N desmethylselegiline with multiple dosing may be the result of binding to the mitochondrial monoamine oxidase type B pool. Given the lack of correlation of the selegiline plasma profile following twice-daily administration with single-dose data, the assessment of bioavailability and meaningful relationships between selegiline plasma levels and pharmacologic response is best studied in a twice daily, multiple-dose setting. PMID- 11862266 TI - Comparative Effects of N(omega)-L-Nitro-L-Arginine Methyl Ester and N(omega)-L Nitro-L-Arginine Benzyl Ester in the Mesenteric Circulation of the Rat. AB - Comparative effects of the nitric oxide (NO) synthase inhibitors, N(omega)-L nitro-L-arginine methyl ester, and N(omega)-L-nitro-L-arginine benzyl ester on baseline arterial tone and on vasodilator responses to acetylcholine, isoproterenol, prostaglandin E(1), histamine, and nitroglycerin were investigated in the isolated mesenteric vascular bed of the rat. Under constant-flow conditions, intra-arterial (IA) injections of acetylcholine (100 ng--1 &mgr;g), isoproterenol (100 ng--1 &mgr;g), prostaglandin E(1) (0.3--3 &mgr;g), histamine (300 ng--3 &mgr;g), and nitroglycerin (100 ng--1 &mgr;g) caused dose-related decreases in mesenteric arterial perfusion pressure and decreases in systemic arterial pressure. Following administration of the NO synthase inhibitors, N(omega)-L-nitro-L-arginine methyl ester or N(omega)-L-nitro-L-arginine benzyl ester, mesenteric vascular resistance, and systemic arterial pressure were increased and mesenteric vasodilator responses to acetylcholine were significantly decreased, whereas N(omega)-L-nitro-L-arginine methyl ester and N(omega)-nitro-L-arginine benzyl ester did not significantly decrease vasodilator responses to nitroglycerin, histamine, isoproterenol, or prostaglandin E(1). The inhibitory effects of N(omega)-L-nitro-L-arginine methyl ester and N(omega)-L nitro-L-arginine benzyl ester on vasodilator responses to acetylcholine suggest that acetylcholine-induced vasodilation in the mesenteric circulation of the rat is dependent on the release of NO from the endothelium. The increase in mesenteric vascular resistance following administration of N(omega)-L-nitro-L arginine methyl ester and N(omega)-L-nitro-L-arginine benzyl ester suggest that tonic production of NO by the endothelium serves to maintain the mesenteric vascular bed of the rat in a dilated state. PMID- 11862267 TI - Atherosclerotic Renovascular Disease. AB - The patient, a 78-year-old Asian male, was brought to the hospital because of acute shortness of breath that had progressively worsened over the course of the day. He complained of a nonproductive cough and claudication after walking 1 block. His past medical history was significant for mild renal insufficiency (serum creatinine 1.5--2.0 mg/dl), the etiology of which was never explored. Although there was a recent history of mild to moderate hypertension, at presentation his blood pressure was noted to be 240/118 mm Hg in both arms. His physical exam at the time of admission was remarkable for grade II hypertensive retinopathy, an S4 gallop, periumbilical systolic bruits, audible femoral arterial bruits and absent distal lower extremity pulses. Initial complete blood count, serum electrolytes and cardiac enzymes (including lactate dehydrogenase) were normal. His blood urea nitrogen and serum creatinine concentrations were 51 and 3.6 mg/dl, respectively, and his urinalysis showed 1+ protein (both by dipstick and sulfasalicylic acid) with a "benign" sediment (0--1 WBC/HPE, 1--2 RBCs/HPF) with occasional granular casts. His electrocardiogram, apart from demonstrating left ventricular hypertrophy with secondary ST-T wave abnormalities, showed no acute changes; his chest X-ray demonstrated cardiomegaly and pulmonary vascular congestion. He was intubated and subsequently treated with increasing parenteral doses of furosemide (40--240 mg) and a nitroglycerine drip (up to 15 mcg/min). Over the course of the first 48 h, his blood pressure was gradually lowered to 170/100 mm Hg. His urine output increased from 20 ml/h to 125/ml/h, and his respiratory status improved, allowing him to be extubated. In spite of adequate control of his blood pressure in the ensuing days (150--170/80- 90 mm Hg), his renal function continued to deteriorate. Renal sonography (without Doppler) demonstrated a right kidney of 9.6 cm and a left kidney of 9.3 cm in length without evidence of hydronephrosis. Both kidneys were noted to be echogenic. Assays for antinuclear antibodies and antineutrophilic cytoplasmic antibodies were negative, and the patient's serum complement levels were normal. For several days after his admission, his serum creatinine gradually rose to 10.7 mg/dl, and hemodialysis was initiated for uremic encephalopathy. Because of the high index of suspicion for renal artery stenosis as the case of both his hypertension and renal failure, a renal angiogram was performed. It revealed a 90% occlusion of the right renal artery with ostial involvement and a 70% occlusion of the left renal artery; both kidneys had poor distal renal vasculature and there was marked atherosclerotic disease of the aorta. After being hemodialyzed for 3 treatments, his renal function began to improve spontaneously. His serum creatinine returned to 3.4 mg/dl, and a subsequent 24 hour urine demonstrated a creatinine clearance of 20 ml/min and an excretion of 1.2 g of protein. Following his discharge from the hospital, his renal function remained unchanged for 3 years, and his blood pressure was easily controlled on monotherapy with a long-acting calcium channel blocker. He recently died from pneumonia. PMID- 11862268 TI - Smoking, Smoking Cessation, and Regulation. PMID- 11862269 TI - Pharmacologic Determinants of Tobacco Dependence. AB - Nicotine is one of the most widely abused psychoactive drugs and far more people trying it progress to regular use than those trying heroin, cocaine, or alcohol. Since the early 20th century, it has been understood that tobacco use is driven largely by nicotine's pharmacological actions. Nicotine produces reinforcing effects, tolerance and physical dependence, and pharmacological effects that smokers enjoy, such as modulation of mood, appetite, and task performance. Pharmacokinetic properties of tobacco-based nicotine products optimize abuse potential. Cigarette smoke inhalation delivers high nicotine concentrations to the brain and other organs within 10 s of inhalation. Nicotine's acute effects dissipate in a few minutes, encouraging the smoker to smoke frequently throughout the day to maintain its pleasurable effects and prevent withdrawal symptoms. Nicotine and other constituents of smoke also provide sensory stimuli which, through repeated pairings, become conditioned reinforcers and further strengthen tobacco self-administration, creating an extremely persistant behavior. Nicotine replacement medications provide relief of cigarette withdrawal symptoms and are effective aids in smoking cessation; however, sustaining long-term abstinence can still be very difficult. PMID- 11862270 TI - Smoking Cessation Strategies. AB - Cigarette smoking is a substantial social and public health problem contributing to about half of the 400,000 cardiovascular deaths in the United States. Nicotine and carbon monoxide are the components in cigarette smoke that contribute to cardiovascular morbidity and mortality. Nicotine is a stimulant of the sympathetic nervous system. It increases myocardial oxygen demand, increases platelet aggregation predisposing to coronary thrombosis and compromises coronary flow, especially in patients with coronary artery disease. Transdermal nicotine in controlled doses has proven to be an effective adjunct to smoking cessation in addition to the nonpharmacological therapy. PMID- 11862271 TI - Smoking, Nicotine, Nicotine Replacement, and Cardiovascular Disease. AB - Cigarette smoking is a major risk factor for the development of coronary artery disease and its complications. Nicotine is one of the major substances that contributes to the pharmacologic activity of cigarette smoke, its toxicity, as well as its addictive nature. Strategies have been developed that utilize nicotine as a replacement substance in motivated patients who wish to stop smoking. Studies have shown that nicotine replacement therapy can be used safely in patients with known coronary artery disease. PMID- 11862272 TI - Smoking Cessation Strategies. PMID- 11862273 TI - Single- and Multiple-Dose Pharmacokinetics of Reduced Metabolite (MDL 74,156) following Oral Administration of Dolasetron Mesylate to Normal Male Subjects. AB - Dolasetron, a 5-hydroxytryptamine(3) receptor antagonist, is under investigation for prevention of nausea and vomiting due to chemotherapy. The keto-reduced metabolite of dolasetron has been identified in human plasma and is likely responsible for the antiemetic activity. This study evaluated single and multiple dose pharmacokinetics of the reduced metabolite following oral administration of dolasetron mesylate in healthy male subjects. Five groups (six active/two placebo each) of subjects received either oral doses of dolasetron mesylate ranging from 25 to 200 mg or placebo on day 1 and every 12 h on days 2 through 9. Because plasma dolasetron concentrations were low and sporadic, pharmacokinetics of the parent compound could not be determined. The reduced metabolite appeared rapidly in the plasma and reached a maximal plasma concentration in about 1 h. The maximal plasma concentrations and areas under plasma concentration--time curves were proportional to the dose. The mean apparent oral clearance ranged from 9.89 to 23.10 ml min(minus sign1) kg(minus sign1). The half-life ranged from 5.20 to 10.80 h. Mean renal clearance and fraction of dose excreted in urine were 0.97 to 3.97 ml min(minus sign1) kg(minus sign1) and 7.47 to 31.9%, respectively. The pharmacokinetics of reduced metabolite appears to be dose independent after single and multiple dosing. PMID- 11862274 TI - Metoclopramide-Induced Vascular Hyperreactivity: A Comparative Study Between Normotensive and Hypertensive Subjects. AB - In the present study, we have evaluated the cardiovascular responses to the cold pressor test (CPT) under intravenous metoclopramide (MTC) treatment (7.5 &mgr;g kg(minus sign1) min(minus sign1)) during a 30-min period and two additional periods, before and after MTC by using 5% glucose solution. There was a vascular hyperreactivity induced by MTC, Delta mm Hg was 21.4/24/1 mm Hg (systolic/diastolic blood pressure [BP]) during MTC versus Delta 17.2/17.9 mm Hg during 5% glucose infusion. In nine hypertensive subjects, Delta mm Hg was 36.8/25.7 during MTC versus Delta 31/24 mm Hg during 5% glucose. Bromocriptine pretreatment (7.5 mg daily for 7 days) antagonized MTC-BP increase during CPT. We conclude the following: (1) MTC induces a greater BP response during CPT which is blocked by using bromocriptine, a known DA(2) dopaminergic agonist. (2) A dopaminergic influence is present during CPT in normotensive and hypertensive subjects. PMID- 11862275 TI - Ethanol Pharmacodynamics at Low Blood Concentrations. AB - Ranitidine has been shown to produce increases in blood alcohol concentration (BAC) after low doses of alcohol. The objective of this study was to reproduce, in a controlled setting, the BACs seen after low oral doses of ethanol in the presence and absence of ranitidine and to assess the effect of these concentrations on cognitive performance. An active control group (0.45 g per kg alcohol) was included as a validation of the methodology used. A randomized, double-blind, placebo-controlled, four-way crossover study was performed in eight healthy males. Subjects received a 20-min intravenous infusion of 0.10, 0.15, or 0.45 g per kg alcohol or placebo. Blood samples were obtained to measure BAC and psychometric effects were assessed over 8 h. A pharmacokinetic model was used to fit simultaneously the BAC--time profiles of all three doses for each subject. Cognitive improvement was assessed using digit symbol substitution, continuous tracking, and divided attention tests. Analysis of variance was conducted in order to compare peak blood alcohol impairment (E(max)) and area under the alcohol impairment--time curve (AUEC) across treatments. Observed median peak BAC (C(max)) for 0.10- and 0.15-g/kg dose groups (median BACs 15.2 and 27 mg/dl, respectively) were very similar to the target C(max) (13 and 26 mg/dl). Analysis of variance of AUEC and E(max) showed difference in impairment measures after the 0.10- and 0.15-g/kg doses. A significant difference in impairment measures between placebo and the active control, 0.45 g/kg (median BAC of 110 mg/dl) was observed, indicating that the methodology was capable of detecting significant psychomotor effects at the legal limit of BAC. Results indicated that the BAC increments from 15 to 27 mg/dl are not associated with significantly impaired performance and, hence, it is unlikely that increases in BAC of this magnitude, such as those caused by ranitidine therapy, are of any clinical relevance. PMID- 11862276 TI - Calcium Channel Blockers and the Gastrointestinal Tract. AB - Calcium ions play an important role in the normal functioning of the gastrointestinal system. The calcium channel blockers appear to affect all the organs of the gastrointestinal tract and may have therapeutic efficacy on esophageal spasm, irritable bowel syndrome, mesenteric vascular insufficiency, dyskinesis of the Sphincter of Oddi, and insulinoma. Adverse effects are limited predominantly to constipation and some aggravation of ethanol-induced ulcer disease. In addition, the drugs continue to serve as important biologic probes for understanding the normal and abnormal functioning of the gastrointestinal tract. PMID- 11862277 TI - Changing the Regulatory Process. PMID- 11862278 TI - Influence of HOE 140, A Bradykinin Receptor Antagonist, in the Isolated Mesenteric Vascular Bed of the Rat. AB - The inhibitory effects of HOE 140 (D-Arg-[Hyp(3),Thi(5),D-Tic(7), Oic(8)]bradykinin), a novel bradykinin B(2)-receptor antagonist, on mesenteric vascular bed vasodilator responses to bradykinin (BK) were investigated under constant-flow conditions in the isolated blood-perfused rat mesenteric vascular bed. During baseline conditions, injections of BK produced dose-related decreases in mesenteric arterial perfusion pressure which were reproducible with respect to time. HOE 140, in a dose of 50 &mgr;g/kg intravenously, decreased vasodilator responses to BK but had no significant effect on mesenteric vasodilator responses to albuterol, acetylcholine, levcromakalim, or to nitroglycerin. These results suggest that BK has significant vasodilator actions which are mediated by the activation of kinin B(2)-receptors in the mesenteric vascular bed of the rat. HOE 140 caused a significant increase in baseline mesenteric arterial perfusion pressure. These data indicate that HOE 140 is a highly selective, BK B(2) receptor antagonist in the mesenteric vascular bed of the rat, and the elevation of baseline mesenteric arterial tone by HOE 140 suggests that BK plays a role in maintaining normal vascular tone in the mesenteric circulation. These results also suggest that HOE 140 is a useful probe for studying the role of BK in the mesenteric vascular bed under physiologic and pathophysiologic conditions. PMID- 11862280 TI - Lithium Withdrawal in Bipolar Disorder: Implications for Clinical Practice and Experimental Therapeutics Research. AB - Recent clinical research strongly suggests that there is a period of elevated risk of morbidity in the several months following abrupt discontinuation or reduction of doses in maintenance treatments commonly used in the contemporary management of chronic or recurring major psychiatric disorders. This risk is best quantified for the discontinuation of lithium therapy in bipolar disorders, in which risk of mania, depression, and suicidal behavior may rise. Similar symptomatic risks are well known after stopping antianxiety agents, and probably also follow rapid removal of oral neuroleptics in schizophenia and antidepressants in major depression. Morbid risk probably can be limited by slowing the rate of removal of medication. Such responses probably reflect interactions of individual illness diatheses with physiological adaptations to long-term drug treatments. The findings appear to have broad scientific, clinical, and ethical implications for clinical pharmacology in psychiatry as well as in general medicine. PMID- 11862279 TI - Effects of Isradipine on 24-h Blood Pressure, Left Ventricular Hypertrophy and Hemodynamics in Type II Diabetic Hypertensive Patients. AB - The effects of 5 mg slow-release isradipine given once daily for both office taken and monitored 24-h blood pressure, left ventricular mass and forearm hemodynamics have been evaluated by noninvasive methods for 6 months in 12 type II diabetic patients aged 40--60 years with concomitant arterial hypertension of mild to moderate degree. A significant reduction in both supine (from 164/96 to 146/85 mmHg) and standing blood pressure (from 164/100 to 143/89 mmHg) was observed at the end of the treatment period. A slight reduction was detected in walking blood pressure and a more marked reduction in sleeping blood pressure measured by an ambulatory blood pressure monitoring system, although the statistical significance was only approached because of the huge daily variability in blood pressure levels. A significant reduction in left ventricular mass index was detected by M-mode echocardiography (from 153 plus minus 72 to 122 plus minus 84 g/m(2) body surface area) without any impairment of left ventricular systolic function. Brachial artery compliance was not significantly increased (from 1.86 plus minus 0.7 to 2.21 plus minus 0.9 cm(4) center dot dyne(minus sign1) center dot 10(7)) and impedance slightly but nonsignificantly reduced (from 99.6 plus minus 52 to 65.7 plus minus 13 dyne(5) center dot cm(minus sign5) center dot 10(2)) by the treatment. These results show that long term isradipine treatment reduces blood pressure values and left ventricular hypertrophy. However, the findings of this study need to be confirmed in a larger population sample in order to establish whether isradipine really improves the overall cardiovascular risk profile. PMID- 11862281 TI - Fluticasone Propionate Reduces Oral Prednisone Use While it Improves Asthma Control. AB - This study examined the effect of fluticasone propionate aerosol on oral prednisone requirements in patients with severe asthma. Ninety-six patients dependent on oral prednisone were treated with placebo or fluticasone propionate aerosol (750 or 1000 &mgr;g twice daily) for 16 weeks. The dosage of oral prednisone was adjusted weekly according to predetermined criteria. Fluticasone propionate 750 and 1000 &mgr;g twice daily resulted in 69% and 88% of patients (low and high doses, respectively) not using any prednisone compared to 3% of placebo-treated patients by the end of the study. In the fluticasone propionate groups, forced expiratory volume in 1 s (FEV(1)) and peak expiratory flow rates and the number of nighttime awakenings improved at the last evaluable visit. In addition, the number of nighttime awakenings and symptomatic albuterol use declined relative to placebo values (p < 0.05). Fluticasone propionate aerosol was well tolerated. Fluticasone propionate aerosol (750 or 1000 &mgr;g twice daily) effectively and safely allowed most asthmatics who were dependent on oral corticosteriods to reduce or eliminate oral prednisone use while improving pulmonary function. PMID- 11862282 TI - Combination Treatment with Sustained-Release Verapamil and Indapamide in the Treatment of Mild-to-Moderate Hypertension. AB - A multicenter study of the treatment of mild and moderate hypertension compared three once-daily regimens for efficacy and safety: Group I, verapamil sustained release (SR) 240 mg; Group II, verapamil SR 480 mg; and Group III, combination therapy of verapamil SR 240 mg plus indapamide 2.5 mg. After a 3-week placebo washout period and a 2-week preliminary treatment period with verapamil SR 240 mg daily, those patients with diastolic blood pressures of <95 mm Hg were excluded from further study; 137 remaining patients with sitting diastolic blood pressure of 95--115 mm Hg, representing "incomplete" responders to verapamil SR 240 mg, were randomized in a double-blind fashion to one of the three treatment groups for a duration of 16 weeks. Efficacy was assessed after 16 weeks of double-blind therapy or at end point. All three treatments significantly reduced sitting diastolic and systolic blood pressure from baseline levels. Compared with Group I (verapamil SR 240 mg daily), there was a significant reduction (p < 0.05) in Group II combination therapy of 8.6 mm Hg systolic and 3.0 mm Hg diastolic and a significant reduction (p < 0.05) in Group II (verapamil SR 480 mg daily) of 7.6 mm Hg systolic and 3.9 mm Hg diastolic BP. Patients who had the least change in blood pressure (diastolic blood pressure decreases of <5 mm Hg) to lead-in verapamil SR 240 mg daily, prior to randomization, tended to have a greater response to combination therapy than to verapamil SR 480 mg. Adverse experiences leading to withdrawal from the study occurred in 21% of patients receiving verapamil SR 480 mg daily (Group II). There was a significantly greater withdrawal from Group II than the other two treatment groups (8.5% from Group III and 4% from Group I). This trial demonstrated that adding indapamide 2.5 mg to the incomplete responders of verapamil SR 240 mg enhances efficacy and was well tolerated. PMID- 11862283 TI - Anaphylaxis and Anaphylactoid Reactions: Diagnosis and Management. AB - Anaphylaxis is an acute fatal or potentially fatal hypersensitivity reaction. Anaphylaxis represent a clinical diagnosis based on history and physical examination and includes symptoms of airway obstruction, generalized skin reactions, particularly flushing, itching, urticaria, angioedema cardiovascular symptoms including hypotension and gastrointestinal symptoms. These symptoms result from the action of mast cell mediators, especially histamine and lipid mediators such as leukotrienes and platelet activating factor on shock tissue. The shock tissue includes blood vessels, mucous glands, smooth muscle, and nerve endings. Anaphylaxis follows the typical immediate hypersensitivity time course, with a reaction beginning within minutes of antigen exposure. A late-phase reaction hours after the initial reaction may occur. Mast cell mediator release can be triggered by both IgE and non--IgE-mediated factors. Therefore, anaphylaxis may be termed anaphylaxis (IgE mediated) or anaphylactoid (non--IgE mediated). The most common IgE-mediated triggers are drugs, typically penicillin or other beta-lactam antibiotics, foods, most commonly nuts, peanuts, fish and shellfish, or hymenoptera stings. Non-IgE-mediated causes include factors causing marked complement activation such as plasma proteins or compounds which act directly on the mast cell membrane, such as vancomycin, quinolone antibiotics, or radiographic contrast media. The pathophysiology of some trigger factors, such as aspirin, remains unclear. Therapy of anaphylaxis revolves around patient education, avoidance, desensitization or pharmacologic pretreatment when agents causing anaphylaxis need to be readministered, and early recognition and prompt therapy of reactions should they occur. PMID- 11862285 TI - Sinusitis: Current Diagnostic and Treatment Strategies. PMID- 11862284 TI - Histamine and Antihistamines: Clinical Pharmacology for the Practicing Physician. PMID- 11862286 TI - Clinical Trials: Design Flaws Associated with Use of a Placebo. AB - BACKGROUND: The placebo arm of a trial, although it controls for psychological placebo effects and natural history of a disease and facilitates effective blinding, is susceptible to bias just like the true treatment arm is. The placebo arm of a trial may even be particularly vulnerable to specific types of bias. The present article focuses on design flaws associated with the use of a placebo. RESULTS: (1) Placebo-controlled trials using mortality rather than morbidity as the end point may have both ethical and statistical problems. (2) Bias may affect placebo data to a larger extent than active treatment data. (3) Trials may provide an atmosphere of exaggerated placebo effects and, consequently, inadequate power to determine differences between placebo and active treatment groups. (4) A biased placebo period due to carryover effect is a common problem of controlled trials with a crossover or self-controlled design. (5) Adjustment for asymmetries between placebo and treatment group in a parallel-group trial is inappropriate if such asymmetries are not completely independent on the main variables of the study. (6) Samples may not be representative when a large number of patients inappreciative of a possible treatment with placebo refuse randomization. CONCLUSIONS: It is emphasized that routinely accounting for design flaws associated with use of placebo may further improve the powerful method of controlled clinical trials. PMID- 11862287 TI - Beta Blockade---Selectivity or Not: A Review of an Old Controversy with New Aspects. PMID- 11862288 TI - Atheroembolic Renal Disease. AB - Cholesterol emboli are being increasingly recognized as an important cause of renal dysfunction in an aging US population. Irregularly shaped atheroemboli typically cause partial obstruction of small renal vessels resulting in ischemia. A vasculitis-like picture often evolves with an inflammatory reaction and giant cell formation. Cholesterol emboli may be temporally related to vascular manipulation, anticoagulant, or thrombolytic drug use. Spontaneous cases have been reported. Patients with cholesterol emboli may present with a spectrum of acute renal failure varying from mild and asymptomatic to life-threatening disease. The differential diagnosis includes radiocontrast nephropathy, endocarditis with left-sided emboli, vasculitis, and thrombotic emboli. The physical examination findings suggestive of cholesterol emboli include extrarenal emboli and livedo reticularis. The urinalysis is typically unremarkable. Some patients have hematuria and/or non-nephrotic proteinuria. Serology and hematology results may suggest an inflammatory-like picture with elevated erythrocyte sedimentation rate, hypocomplementemia, eosinophilia, and eosinophiluria. In the setting of a clear precipitating factor and suggestive physical findings, cholesterol emboli can be established purely on clinical grounds. Demonstration of cholesterol crystals by biopsy of the kidney, skin (if lesion present), or muscle is diagnostic in unexplained cases. The kidney is the organ most frequently involved in this order. Therapy is supportive with particular emphasis on management of hypertension and hypercholesterolemia. PMID- 11862289 TI - The Commercialization of Research. PMID- 11862290 TI - Pharmacokinetic and Metabolism Study in Healthy Volunteers After Administration of Single Oral Dose of (3)H-alpha-Dihydroergocryptine Mesylate. AB - This phase I open pharmacokinetic and metabolism study was conducted with six healthy male volunteers who were given 20 mg of (3)H-alpha-dihydroergocryptine in order to evaluate the absorption, plasma time course, and urinary and fecal elimination of total radioactivity. Rapid absorption into the general circulation occurred with an average K(01) of 0.99 plus minus 0.73/h. Peak time(T(max)) was reached in approximately 3 h with an average radioactivity concentration (C(max)) of 8.78 plus minus 5.9 ng eq h/ml. Distribution from the central compartment to the peripheral compartment occurred with a mean rate constant (K(12)) of 0.330 plus minus 0.22/h. Estimations of total clearance (CL) and volume of distribution (Vd) seem strongly affected by the low oral availability (F) of hydrogenated ergots. The rate constant (K(21)) of radioactivity washout from the tissue to the central compartment was 0.250 plus minus 0.130/h. However, plasma radioactivity declined biexponentially with an overall elimination constant (K(10)) of 0.029 to 0.146/h (i.e, half-lives of 23.9--4.75/h). Total radioactivity recovery in urine and feces was good with 82.78 plus minus 6.44% of dose eliminated in feces and 3.01 plus minus 0.65% in urine. The latter concentration was too low to detect metabolites or unchanged drug by radioactivity image scanning. However, the liquid scintillation count of silica gel that had been scraped off the thin layer chromatography (TLC) plates indicated the presence of metabolites in urine. Pharmacodynamically, both supine and standing blood pressure fell significantly within the first 8 h of dosing, yet there were no changes in heart rate. No adverse reactions were reported. In conclusion, the kinetics of (3)H dihydroergocryptine are very similar to other ergot alkaloids in respect to extensive hepatic metabolism with an elimination half-life of 25 h. PMID- 11862291 TI - Increased Deoxycytidine Kinase mRNA Level After Treatment with Interleukin-3. AB - Pretreatment of IL-3 to Kasumi-1 human acute myeloid leukemia cells enhanced 1-B D-arabinofuranosyl cytosine (ara-C) cytotoxicity 1.2. to 1.4-fold (median 1.3). To clarify the mechanism of interleukin-3 (IL-3) on ara-C cytotoxicity, we investigated the level of deoxycytidine kinase mRNA with the competitive polymerase chain reaction method and enzyme activities, the incorporation of [(3)H] ara-C into DNA and intracellular ara-cytidine triphosphate (CTP) levels with high-performance liquid chromatography and analyzed cell cycles. The level of deoxycytidine kinase mRNA showed a fourfold increase (88.3 plus minus 4.33 amol &mgr;g of total RNA) at 3 days after treatment with IL-3 compared to control (20.3 plus minus 4.33 amol &mgr;g). After IL-3 treatment, ara-C incorporation into the DNA was increased to 1.33 to 1.83-fold (median, 1.73-fold). The G0/G1 late-phase and S-phase percentages of cells were increased from 28.99 to 78.73% in the IL-3 treatment group as compared to control. These results indicate that IL-3 pretreatment increases the level of deoxycytidine kinase mRNA and ara-C incorporation into the DNA and also increases ratios of G0/G1 late-phase and S phase subsequent to an enhancement of ara-C cytotoxicity against leukemia cells. PMID- 11862292 TI - Inhibitory Effects of LY301875 on Responses to Angiotensin Peptides in Pulmonary Vascular Bed of the Cat. AB - The effects of the nonpeptide angiotensin receptor antagonist LY301875 on responses to angiotensin II, angiotensin III, and angiotensin IV were investigated in the pulmonary vascular bed of the intact cat chest. Under conditions of controlled blood flow, injections of the angiotensin peptides into the perfused lobar artery caused dose-related increases in lobar arterial pressure, and LY301875 decreased pressor responses to angiotensin II, angiotensin III, and angiotensin IV. The duration of the blockade was related to the dose of the antagonist, and LY301875 had no significant effect on pressor responses to U 46619, norepinephrine, serotonin or BAY K 8644. LY301875 caused minimal decreases on mean baseline systemic arterial pressures and did not significantly affect lobar arterial pressure in the cat. These results indicate that LY301875 is a potent selective long-acting angiotensin receptor antagonist and suggest that this agent may be useful in the investigation of the role of angiotensin peptides in physiological and pathophysiological processes in the pulmonary vascular bed. PMID- 11862293 TI - Serum Estradiol and Estrone Levels During 3 Weeks of Alternate-Site and Same-Site Applications of a Once-A-Week Drug-in-Adhesive Transdermal Estradiol Patch. AB - This open-label, randomized crossover trial was conducted in healthy postmenopausal women to characterize the serum estradiol and estrone profiles from a once-a-week transdermal estradiol patch applied repeatedly to the same site or to alternate sites on the abdomen over a 3-week period. Similar mean serum estradiol and estrone profiles and circulating estradiol:estrone ratios of approximately 1.0 were observed for both patch application regimes. Repeated application to the same or alternate sites did not result in an accumulation of exogenous estradiol or in a statistically significant increase in application site reactions. Although the once-a-week transdermal estradiol patch should be applied to alternate sites to reduce the likelihood of dermatologic reactions, accidental application site overlap should not result in an increase in serum estradiol and estrone concentrations. PMID- 11862294 TI - Bioequivalence Comparison of Two Drug-in-Adhesive Transdermal Nitroglycerin Patches. AB - Plasma concentrations of nitroglycerin, 1,2-dinitroglycerin and 1,3 dinitroglycerin in 50 healthy male subjects who received single 24-h applications of two 0.6 mg/h drug-in-adhesive transdermal nitroglycerin patch formulations were compared for the purpose of establishing bioequivalence. The rates of appearance and elimination of nitroglycerin and both dinitro metabolites were the same for both patch treatments. The pharmacokinetic and statistical analyses indicated that the 20-cm(2) test patch met the current two one-sided t-test criteria for bioequivalence with the 30-cm(2) reference patch for nitroglycerin and both dinitro metabolites, even though the reference patch was 50% larger in size and contained 120% more nitroglycerin than the test patch. A likely explanation for the equivalence in drug delivery with a smaller patch is that the test patch maintained more intimate skin contact than did the reference patch. This study shows that strict bioequivalence criteria can be met with a highly variable drug such as nitroglycerin. PMID- 11862295 TI - New Therapeutic Approaches to Peptic Ulcer Disease: The Role of Helicobacter Pylori. AB - One of the most common bacterial infections of human involves Helicobacter pylori, a spiral, gram-negative bacterium that is now thought to be a dominant factor in the development of peptic ulcer disease and may be significant in causing certain forms of gastric cancer. Almost 100% of patients with duodenal ulcer and 70 to 90% affected with gastric ulcer are infected with H. pylori. In order to achieve cure of H. pylori--induced ulcer disease, it is necesary to eradicate the bacterial infection. Mere suppression or clearance infection without eradication is associated with a >80% recurrence of the ulcer. The epidemiology, microbiology, and pathogenesis of H. pylori infections are reviewed. Diagnostic methods and optimal treatment strategies for H. pylori infections are examined. The most current diagnostic and treatment algorithms for peptic ulcer disease are discussed critically, and future directions for drug development aimed at eradication of H. pylori infection are considered. PMID- 11862296 TI - Saterinone: A New Dual-Action Drug in the Acute Management of Chronic Heart Failure. AB - A new dual-action drug called saterinone combines both alpha-1 blocking vasodilatory property and phosphodiesterase III inhibition--mediated inotropism. A placebo-controlled, randomized, double-blind study was performed in 12 patients with severe congestive heart failure. Either 2 &mgr;g center dot kg center dot min(minus sign1) saterinone (n = 8) or placebo (n = 4) was injected intravenously over 3 h at rest. On-line hemodynamic measurement utilizing intra-arterial blood pressure monitoring and two-dimensional (2D) echocardiography were performed at basal time; then 30, 60, 120 and 180 min after infusion. The parameters measured were blood pressure (mmHg), systemic vascular resistance (SVR dynes center dot cm(5) center dot min(minus sign1)), pulmonary artery pressure (PAP mmHg), mean pulmonary capillary wedge pressure (PCWP mmHg) and cardiac index (CI 1 min/m(2)) using right heart catheterization and end-systolic volume (ESV ml) and end diastolic volumes (EDV ml), ejection fraction (EF%) using 2D echocardiography. Placebo had no significant effects on any of the parameters (p = NS). Saterinone decreased SVR by 37% (p < 0.001), PAPm by 24% (p < 0.05), PCWP by 35% (p < 0.05), ESV by 27% (p < 0.01) and increased CI by 32% (p < 0.05) and EF by 45% (p < 0.05). Saterinone appears to be a potent drug that produces improvements in both cardiac hemodynamics and LV functional parameters. Further study with this interesting agent is indicated. PMID- 11862297 TI - Therapeutic Commentary: Dexfenfluramine. PMID- 11862298 TI - What Data Are Required to Adopt New Antihypertensive Agents? AB - The arguments presented in this brief review indicate that despite the large number of antihypertensive agents currently available, there is still need for additional drugs. Introduction of new members of an existing class is justified if the new drug presents advantages in terms of ancillary properties or side effect profile, when compared with other agents of that class. Novel classes of antihypertensives, with a mechanism of action different from existing agents, such as the recently introduced angiotensin II antagonists, are of particular interest because they affect a different pathogenetic aspect of the disease. The efficacy of new drugs should be tested by comparison with established drugs, since placebo controls are no longer justifiable except for short periods. Longitudinal comparisons of drugs from different classes are necessary to establish their relative potential for protection of vital organs from hypertensive damage. PMID- 11862299 TI - The FDA and Software. PMID- 11862300 TI - Comparison of Plasma Protein Binding of Basic Drugs in Black and White Individuals. AB - Interethnic difference in drug disposition is an important contributing factor to interindividual variation in drug response. Since interethnic differences in the protein binding of drugs may contribute to variation in drug disposition between ethnic groups, we conducted a study in 10 black Americans (A) and mean (plus minusSE) age 26 plus minus 6 years and weight 80 plus minus 9 kg matched against 10 white Americans (C) with a mean age of 28 plus minus 6 years and weight 81 plus minus 9 kg, all within 10% of ideal body weight. Serum alpha-1-acid glycoprotein (AGP) and albumin concentrations were measured using the auramine-O and bromcresol green methods, respectively. Verapamil, propranolol, lidocaine, disopyramide and diazepam binding in plasma were measured with the equilibrium dialysis method, involving the determination of free and unbound drug concentrations. The unbound fraction of diazepam (A = 1.1 plus minus 0.1%; C = 1.1 plus minus 0.1%), verapamil (A = 9.5 plus minus 0.8%; C = 9.8 plus minus 0.4%), propranolol (A = 14.2 plus minus 1.0%; C = 12.6 plus minus 0.7%), lidocaine (A = 28.5 plus minus 2.1%; C = 25.7 plus minus 1.1%) and diphenhydramine (A = 42.9 plus minus 10.2; C = 30.4 plus minus 7.01%) showed no significant ethnic differences (unpaired t-test). Disopyramide measured at 7 different concentrations (1.0--20.0 &mgr;g/ml) was similar in both groups, as were the plasma concentrations of AGP (A = 100 plus minus 20 mg 100 ml; C = 120 plus minus 20 mg 100 ml) and albumin (A = 4.3 plus minus 0.1 g 100 ml; C = 4.5 plus minus 0.1 g 100 ml). It is therefore concluded that there are no interethnic differences in the protein binding of basic drugs between black Americans and white Americans and that it is not a major contributing factor to any possible interethnic variation in the disposition of responsiveness of these drugs. PMID- 11862301 TI - Effects of Terazosin on Glycemic Control, Cholesterol, and Microalbuminuria in Patients with Non--Insulin-Dependent Diabetes Mellitus and Hypertension. AB - The use of antihypertensive agents that have positive or neutral effects on blood sugar, lipid profiles, and microalbuminuria can be important clinical treatment for patients with diabetes. We evaluated the effects of both terazosin, a selective alpha-one-adrenergic blocker, and hydrochlorothiazide (HCTZ), a standard mild antihypertensive agent, on glycemic control, urinary albumin excretion rate overnight total cholesterol, and other parameters in non--insulin dependent diabetes mellitus (NIDDM) patients. A randomized, placebo-controlled, cross-over design was implemented in 25 patients. Over an 8-week treatment period fasting plasma glucose (FPG) and glycosylated hemoglobin (GHgb) improved in the terazosin group. Post-treatment FPG was 200 plus minus 85 and 187 plus minus 71 for patients who received HCTZ and terazosin, respectively. Although the GHgb improved significantly for terazosin patients (12.2 plus minus 5.8 for HCTZ versus 10.7 plus minus 4.6 for terazosin, p = 0.03), microalbuminuria did not improve in terazosin patients in this pilot study. A larger randomized study with tighter blood pressure end points are needed to assess fully the impact of terazosin on micoroalbuminuria and overall glycemic control in the NIDDM patient. PMID- 11862302 TI - Clinical Comparison of Cefuroxime Axetil with Cefixime in the Treatment of Acute Bronchitis. AB - Acute bronchitis is the ninth most common outpatient illness seen by physicians in the United States. Oral antibiotic treatment is usually directed empirically against the most common bacterial pathogens associated with acute bronchitis, such as Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis. Although cefuroxime axetil and cefixime are both approved in the United States for treatment of acute bronchitis, currently they have not undergone direct clinical comparison for this indication. This randomized, investigator-blind, multicenter study was designed to compare the efficacy and safety of 250 mg cefuroxime axetil administered twice daily with that of 400 mg cefixime administered once daily in the treatment of acute bronchitis. Outpatients had to be greater-than-or-equal12 years of age and have signs and symptoms of acute bronchitis to be eligible for this study. Patients were randomly assigned to receive 10 days of oral treatment with either 250 mg cefuroxime axetil taken twice daily or 400 mg cefixime taken once daily. Patients were assessed for both clinical and bacteriologic responses once during treatment (3--5 days) and twice after treatment (1--3 days and 14 days). Bacteriologic assessments were based on sputum specimen cultures obtained pretreatment and posttreatment when possible. Of 465 patients with acute bronchitis who were enrolled in the study, 227 received cefuroxime axetil and 238 received cefixime. Organisms were isolated from the pretreatment sputum specimens in 172 of the 465 (37%) patients, with the primary pathogens being Haemophilus influenzae, Streptococcus pneumoniae, Morazella catarrhalis, and Staphylococcus aureus (30%, 14%, 14%, and 14% of isolates, respectively). A satisfactory clinical outcome (cure or improvement) was achieved in 88% (130 of 148) and 91% (152 of 167) of the clinically evaluable patients who had received cefuroxime axetil or cefixime, respectively (p = 0.36). Regarding the eradication of bacterial pathogens, a satisfactory outcome (cure or presumed cure) was obtained in 89% (47 of 53) and 91% (41 of 45) of bacteriologically evaluated patients who had received cefuroxime axetil or cefixime, respectively (p = 0.75). Treatment with cefixime was associated with a significantly higher incidence of drug-related gastrointestinal adverse events than was treatment with cefuroxime axetil (18% versus 10%, respectively; p = 0.01). This difference primarily reflects a higher incidence of drug-related diarrhea (15% versus 5%, p = 0.001). These results indicated the cefuroxime axetil taken twice daily is as effective as cefixime taken once daily in the treatment of acute bronchitis and that cefuroxime axetil produces fewer gastrointestinal adverse events, particularly diarrhea. PMID- 11862303 TI - Pharmacokinetic Study of Triflusal in Elderly Subjects After Single and Repeated Oral Administration. AB - In this study the single-dose and steady-state pharmacokinetics of unchanged triflusal and its metabolite 2-hydroxy-4-trifluoromethylbenzoic acid (HTB) were studied in 12 elderly subjects treated with a single oral administration of 300 mg triflusal and repeated oral administrations of 300 mg triflusal b.i.d. for 13 days. After a single administration, unchanged triflusal is promptly absorbed (t(max) 0.75 h, C(max) 3.83 &mgr;g/mL) and rapidly depleted from the systemic circulation. Its concentration was measurable only up to 1 to 4 h after administration. The apparent terminal half-life was 0.85 h. HTB proves to be quickly generated from triflusal (t(max) 2.00 h, C(max) 39.88 &mgr;g/mL) and slowly eliminated from the body (t = 54.6 h). With the dose regimen proposed, unchanged triflusal does not accumulate in the body. Conversely, HTB plasma concentration builds up progressively toward steady-state levels of approximately 102 &mgr;g/mL after 4 to 5 d of treatment. No substantial change in peak time, elimination rate constant and half-life evaluated after single-dose treatment was observed on multiple-dose regimen for unchanged triflusal and its metabolite HTB. Therefore, our findings do not indicate a time-dependent pharmacokinetics for triflusal. There were no changes in blood pressure, heart rate or laboratory safety date, i.e., biochemical or hematological profiles. PMID- 11862304 TI - Effect of Low-Dose Niacin on Glucose Control in Patients with Non--Insulin Dependent Diabetes Mellitus and Hyperlipidemia. AB - BACKGROUND: High doses of niacin have been shown to impair glucose control in patients with non--insulin-dependent diabetes mellitus (NIDDM). We undertook a study to determine if low-dose niacin has a similar effect. METHODS: Patients with stable NIDDM and hyperlipidemia underwent a 2-month observation phase where diet and diabetes medication were unchanged. Fasting blood sugar was determined every 2 weeks, and hemoglobin A(1c) and lipid profile was obtained at week 8. Patients then received niacin 500 mg three times daily for 2 months with fasting blood sugar measured every 2 weeks and hemoglobin A(1c) and lipid profile determined at week 16. Statistical analysis was performed using a t-test for related groups. RESULTS: Six of nine patients completed the protocol. Mean fasting blood sugar was statistically higher during niacin therapy versus baseline (131 mg/dL plus minus 27 vs. 161 mg/dL plus minus 40, p < 0.05). Two patients had an increase in fasting blood sugar exceeding 200 mg/dL. No change was noted in hemoglobin A(1c). There was a trend in a decrease in total cholesterol, low-density lipoprotein and triglyceride. High-density lipoprotein was statistically higher after niacin therapy. CONCLUSION: Low-dose niacin increases fasting blood sugar in patients with stable NIDDM. PMID- 11862305 TI - Analysis of Responses of Pentoxifylline in the Pulmonary Vascular Bed of the Rat. PMID- 11862307 TI - Clinical Therapeutic Conference: Therapeutic Considerations in Atrial Fibrillation Management. PMID- 11862306 TI - A New Look At United States Drug Development and Approval Times. AB - Clinical development and regulatory approval times for new chemical entities (NCEs) approved in the United States through 1995 were analyzed by both year of new drug application (NDA) approval and year of NDA submission. Results and conclusions from a recent General Accounting Office (GAO) report on US new drug approval times by year of NDA submission for NDAs submitted during 1987 to 1992 were examined and the analysis was extended using data collected by the Tufts Center for the Study of Drug Development (CSDD). The hypothesis that approval times declined by year of NDA submission for 1987 to 1992 submissions, independent of such factors as the therapeutic type and significance of the drugs reviewed, is not supported by the evidence. Mean times from first testing in humans, investigational new drug application (IND) filing, and initiation of Phase III testing to NDA submission increased over time for NCEs by period of NDA submission. When analyzed by year of approval, mean approval times for the 1994 and 1995 NCE approvals were markedly lower than mean approval times for previous years (1.7 y for 1994--1995 compared to 2.7 years for 1990--1993). Mean time from IND filing to NDA submission, however, was notably longer for the 1994 and 1995 approvals than for approvals in earlier years (7.2 y for 1994--1995 compared to 5.5 y for 1990--1993). PMID- 11862308 TI - Alternative Mechanical Methods of Cardiopulmonary Resuscitation. AB - Cardiopulmonary resuscitation (CPR) has become the standard of care for providing circulatory support to victims of cardiac arrest. However, despite its widespread acceptance, few victims ever survive to hospital admission and even fewer to hospital discharge. In hopes of improving survival rates of those suffering cardiac arrest, investigators have suggested and tested several modifications to standard CPR to optimize its hemodynamic effects. In this review, we discuss the important chest compression techniques that have been used over the past 3.5 decades which have mirrored our evolving understanding of the mechanisms underlying CPR. PMID- 11862309 TI - General Clinical Research Centers in the United States make a healthy recovery. PMID- 11862310 TI - A few comments on the study of excitomotor nervous system of the heart. 1866. PMID- 11862311 TI - Centenary Nobel Prize in Physiology or Medicine for the cell cycle. PMID- 11862312 TI - Linking molecular and bedside research: designing a clinical research infrastructure. AB - For 40 years the General Clinical Research Centers (GCRCs) have provided the infrastructure in Academic Medical Centers (AMCs) in the United States, which enabled and facilitated the extraordinary achievements of twentieth century patient-oriented research. The GCRC program in the United States represents a central theme linking molecular and clinical research directed at elucidating mechanisms of human disease. The GCRC can provide the necessary infrastructure and critical intellectual mass to reach a deeper understanding of human biology and to bring better health to persons in both developed and developing countries. A well-organized GCRC program should encompass educational as well as scientific components. It must be developed with an appropriate quality control infrastructure to provide physician-scientists a broad base of training in ethical principles in conduct of research and in scientific design and modern technology. The Human Genome Project has brought unprecedented opportunities to clinical investigators, but taking full advantage of them will require a rebirth of the GCRC as a program capitalizing on local strengths and differences, a more multicentric program, and a program reaching outside the GCRC walls, beyond institutional walls, and into the community. The GCRCs can appropriately be the clinical arm of the human genome project, harnessing the genetic revolution for human health. The GCRC model can provide a conceptual base on which new international patient-oriented research infrastructures can develop. PMID- 11862313 TI - Nucleic acid based strategies as potential therapeutic tools: mechanistic considerations and implications to restenosis. AB - The capacity of DNA to bind RNA via Watson-Crick base-pairing is fundamental to antisense oligonucleotide strategies to inhibit gene expression, and is a property that has been exploited by bioengineers in the generation of catalytic molecules such as ribozymes, ribozyme subtypes, and more recently DNAzymes. This review describes the evolution of these gene-specific agents and summarizes recent efforts to inhibit smooth muscle cell growth with these molecules as candidate therapeutic tools in restenosis. PMID- 11862314 TI - Thyrotropin receptor mutations as a tool to understand thyrotropin receptor action. AB - A large number of mutations have been identified in the thyrotropin (TSH) receptor (TSHR) gene causing human diseases. Toxic thyroid nodules are frequently associated with somatic constitutively activating TSHR mutations. Autosomal dominant non-autoimmune hyperthyroidism is caused by activating TSHR germline mutations. Inactivating germline mutations cause TSH unresponsiveness. Discovery of the different TSHR mutations in various regions of the receptor molecule has led to the identification of important domains for intramolecular TSHR signal transduction. However, despite the functional characterization of the naturally occurring mutations the precise molecular mechanisms of receptor activation including the processes of hormone binding, intramolecular signaling between the different TSHR domains and of G protein coupling are not completely understood. This review discusses the importance of the various receptor domains for TSHR activation identified on the basis of the naturally occurring gain or loss of function mutations and in vitro investigations performed with site-directed mutagenesis, synthetic peptides, or antibodies. Several in vitro studies have provided new insights into structure-function relationships by site-directed mutagenesis in combination with molecular modeling. These in vitro investigations have often been guided by naturally occurring mutations and have provided new insights into intramolecular changes during receptor activation. This has led to progress in understanding the mechanism of TSHR activation. PMID- 11862315 TI - A population of autoantibodies against a centromere-associated protein A major epitope motif cross-reacts with related cryptic epitopes on other nuclear autoantigens and on the Epstein-Barr nuclear antigen 1. AB - Autoimmune diseases arise from a host's immune response against self-antigens. The triggering events ultimately resulting in such a break of tolerance are largely unknown. It is also not known why certain molecular structures become autoantigenic. The hypothesis has long been proposed that autoimmune diseases arise from molecular mimicry followed by an epitope spreading mechanism. Recently we have shown that the anti-centromere-associated protein A (CENP-A) immune response is directed against an autoantigenic motif, G/A-P-R/S-R-R, that occurs three times in the N-terminal amino acids of CENP-A. In the present study we used mutational analyses with immobilized oligopeptide arrays to identify the amino acids in this motif that are responsible for antibody binding. In particular, we found that surprisingly mimotopes of this motif are present in a vast number of autoantigens and in the Epstein-Barr nuclear antigen 1. With affinity-purified antibodies we show that the antibodies against this motif are polyclonal and cross-react with several autoantigens. However, in these autoantigens this motif often represents a cryptic epitope explaining the obvious conflict between our results and the known high specificity of autoantibodies. The presence of such an ubiquitous structure on autoantigens suggests a novel peptide-driven mechanism for the evolution of autoantibodies. PMID- 11862317 TI - PPARgamma and insulin sensitivity: too much and too little of a good thing. PMID- 11862316 TI - Apolipoprotein E gene promoter (-219G/T) polymorphism is associated with premature coronary heart disease. AB - The relationship of two apolipoprotein (apo) E gene polymorphisms and coronary heart disease (CHD) was investigated in 118 Finnish families with premature CHD and in 110 healthy control subjects. Affected siblings and probands with premature CHD had higher frequencies of the T allele of the -219G/T promoter polymorphism and the epsilon 4 allele (genotypes epsilon 4/3 or epsilon 4/4) of the apo epsilon 2/epsilon 3/ epsilon 4 polymorphism than those of healthy control subjects. Additionally, when the two apo E gene polymorphisms were combined, affected siblings and probands had a higher frequency of the -219T allele and the epsilon 4 allele combinations than did healthy controls. The -219T and the epsilon 4 alleles both separately and together were associated with higher levels of 2-h glucose in an oral glucose tolerance test. These results indicate that the two polymorphisms of the apo E gene have similar effects on the risk of coronary atherosclerosis in families with premature CHD. This risk was not explained by the effect of apo E gene polymorphisms on cholesterol metabolism, but their effect on cardiovascular risk factor clustering with insulin resistance may be of importance. We conclude that in addition to the epsilon 4 allele, also the 219G/T promoter polymorphism of the apo E gene is associated with early onset CHD. PMID- 11862318 TI - Further reports on diabetes mellitus after the exstirpation of the pancreas. PMID- 11862319 TI - Prostaglandins as inflammatory messengers across the blood-brain barrier. AB - Upon immune challenge the brain launches a wide range of responses, such as fever, anorexia, and hyperalgesia that serve to maintain homeostasis. While these responses are adaptive during acute infections, they may be destructive during chronic inflammatory conditions. Research performed during the last decade has given us insight into how the brain monitors the presence of a peripheral inflammation and the mechanisms underlying the brain-mediated acute-phase reactions. Here we give a brief review on this subject, with focus on the role of prostaglandin E2 produced in cells associated with the blood-brain barrier in immune-to-brain signaling. The recent advances in this field have not only elucidated the mechanisms behind the anti-pyretic and anti-hyperalgesic effects of cyclooxygenase inhibitors, but have also identified novel and more-selective potential drug targets. PMID- 11862320 TI - Vasoactive intestinal peptide in the immune system: potential therapeutic role in inflammatory and autoimmune diseases. AB - Vasoactive intestinal peptide (VIP), a neuropeptide that is produced by lymphoid as well as neural cells, exerts a wide spectrum of immunological functions, controlling the homeostasis of the immune system through different receptors expressed in various immunocompetent cells. In the last decade, VIP has been clearly identified as a potent anti-inflammatory factor, which acts by regulating the production of both anti- and pro-inflammatory mediators. In this sense, VIP has been described to prevent death by septic shock, an acute inflammatory disease with a high mortality. In addition, VIP regulates the expression of co stimulatory molecules, this being an action that may be related to modulating the shift toward Th1 and Th2 differentiation. We have recently reported that VIP prevents the deleterious effects of an experimental model of rheumatoid arthritis, by downregulating both inflammatory and autoimmune components of the disease. Therefore, VIP has been proposed as a promising candidate alternative treatment for acute and chronic inflammatory and autoimmune diseases such as septic shock, arthritis, multiple sclerosis, Crohn disease, or autoimmune diabetes. PMID- 11862321 TI - Suppression of mismatched mutation by p53: a mechanism for guarding genomic integrity. AB - The tumor suppressor p53 plays an important role in guarding the genomic integrity of the cells. The 3'-->5' exonuclease activity of p53 has recently been recognized as a novel biochemical function of this molecule, and has been shown to preferentially excise mismatched nucleotides from DNA and enhance the DNA replication fidelity of polymerase alpha in vitro. The present study further investigated the role of this biochemical function in whole cells by testing the possibility that p53 may reduce mismatched mutations in cells under a stress of DNA replication errors. Cells with different states of p53 expression, either endogenously or ectopically, were exposed to hydroxyurea to induce an imbalance of cellular dNTP pools and cause replication errors. The rates of mutation at the hypoxanthine guanine phosphoribosyltransferase ( HPRT) gene were determined by selecting colonies of HPRT- mutants. Incubation of cells with hydroxyurea induced a similar degree of dNTP pool imbalance in each cell line, but caused significantly more mutations in cells lacking p53 protein expression. The mutation frequency was significantly reduced by introduction of a wild-type p53 expression vector into the p53-null cells. Analysis of the mutants demonstrated that the clones were devoid of HPRT enzyme activity, but appeared to transcribe full-length HPRT mRNA. These data suggest that p53 is able to reduce mutations caused by misincorporation of deoxynucleotides. Thus, the preferential removal of mismatched nucleotides from DNA by p53 may be a mechanism to maintain genomic integrity. Defect in this biochemical function of p53 may contribute to genetic instability associated with cancer development and progression. PMID- 11862322 TI - Interaction effect between common polymorphisms in PPARgamma2 (Pro12Ala) and insulin receptor substrate 1 (Gly972Arg) on insulin sensitivity. AB - The Pro12Ala polymorphism in the peroxisome proliferator-activated receptor (PPAR) gamma2 gene is associated with a reduced risk of type 2 diabetes. A beneficial effect on insulin sensitivity is reported in some but not all populations. It is possible that this genetic variant produces a characteristic phenotype only against a certain genetic background. We therefore tested the hypothesis that carriers of the Ala allele of PPARgamma2 exhibit a different phenotype against the background of the Gly972Arg polymorphism in the insulin receptor substrate (IRS) 1. We determined insulin sensitivity in the four combinations defined by the absence or presence of the polymorphic allele (healthy, glucose tolerant subjects), by the oral glucose tolerance test (OGTT; using a validated index, n=318) and hyperinsulinemic clamp ( n=201). Insulin sensitivity was not or was only marginally different between Pro/Pro and X/Ala in the overall population. Interestingly, using the OGTT index, insulin sensitivity was significantly greater in X/Ala (PPARgamma2) + X/Arg (IRS-1) than in Pro/Pro (PPARgamma2) + X/Arg (IRS-1). On the other hand, insulin sensitivity was similar in the X/Ala (PPARgamma2) + Gly/Gly (IRS-1 972) and the Pro/Pro (PPARgamma2) + Gly/Gly (IRS-1). The results were practically identical using insulin sensitivity from the clamp. In conclusion, the Arg972 (IRS-1) background produced a marked difference in insulin sensitivity between X/Ala and Pro/Pro (PPARgamma) which was not present in the whole population or against the Gly972 (IRS-1) background. This suggests that the Ala allele of PPARgamma2 becomes particularly advantageous against the background of an additional, possibly disadvantageous genetic polymorphism. Allowing for gene-gene interaction effects may reveal novel information regarding metabolic effects of genetic variants. PMID- 11862323 TI - Glutathione- S-transferase micro and theta gene polymorphisms as new risk factors of atopic bronchial asthma. AB - The genetic polymorphism of glutathione- S-transferase M1 (GSTM1) and glutathione S-transferase T1 (GSTT1) genes and the cytochrome P4501A1 gene responsible for xenobiotic conjugating enzymes of the phase II and phase I detoxification system were studied by PCR-RFLP in the blood spots of 109 patients with atopic bronchial asthma and 90 healthy individuals. GSTM1 gene deletion (GSTM1(0/0)) was detected in 47.8% of individuals in the control group and in 76.1% of asthmatic patients. Individuals without the GSTM1 gene were at approximately 3.5--fold higher risk of developing asthma. The proportion of GSTT1(0/0) genotypes was significantly higher in the group of asthmatics (67.0%) than in controls (23.3%). The proportion of individuals with a deficiency in both GSTM1 and GSTT1 gene activity was more than four times higher in asthmatic patients than in the control group (54.1% and 12.2%, respectively). The frequency of the Ile-Val polymorphism of the CYP1A1 gene was similar in controls and asthmatic patients. This study shows the association of atopic bronchial asthma with GSTM1(0/0), GSTT1(0/0) genotypes. PMID- 11862324 TI - Heterogeneity of TT virus related sequences isolated from human tumour biopsy specimens. AB - TT viruses have recently been reported in serum samples from varying percentages of human blood donors and in patients with chronic liver disease. However, no association with human pathology has yet been reported. In a pilot study we analysed 162 biopsy specimens from various human cancers and from colon polyps for the presence of TT virus related sequences by polymerase chain reaction using three sets of nested primers for TT virus detection. All gels were subjected to Southern blot hybridisation, and DNA from hybridising bands was cloned and sequenced. A total of 54.3% of tumour specimens contained identifiable TT virus related sequences. Specimens from hypopharynx, larynx, endometrial, ovarian and bladder cancers were 14-35% positive and gastrointestinal cancers (oesophagus, stomach, colon, rectum) and colon polyps 57-100% positive. Lung cancers (68.4%), mammary cancers (50%), multiple myelomas (85.7%) and human leukaemias (53.3%) also revealed a high prevalence of TT virus related sequences. Since normal control tissues were not available for the tumour biopsy specimens tested, these data do not permit conclusions concerning a possible causal relationship between the virus infections and carcinogenesis. Another aspect, however, deserves attention: the heterogeneity of TT virus clones obtained from the specimens tested here was striking: 66 novel sequences, probably belonging to new types were identified. Only 16 clones corresponded by more than 97% of their sequences to established prototypes. Even individual tumours commonly contained sequences substantially diverging in nucleic acid composition. Up to five different types were identified within an individual tumour. The high variability of these virus types suggests that additional primer combinations within the highly conserved region of the genome would detect a still higher rate of positive tumours. PMID- 11862326 TI - Proliferation-dependent induction of apoptosis by the retinoid CD437 in p53 mutated keratinocytes. AB - 6-[3-(1-adamantyl)-4-hydroxyphenyl]-2-Naphthalene carboxylic acid (CD437) is a synthetic retinoid with strong apoptogenic properties in various neoplastic cell lines. CD437 was shown to induce apoptosis in malignant human keratinocytes but not in normal keratinocytes. We demonstrate that CD437 is also capable of inducing apoptosis in the non-tumorigenic keratinocyte cell line HaCaT that carries UV-type mutations on both alleles of the p53 gene. The concentration dependent induction of apoptosis was restricted to proliferative HaCaT cells, whereas no effect was seen in differentiating post-mitotic cells. The apoptotic elimination of the proliferative cells was accompanied by rapid upregulation of c jun, downregulation of c- fos, and activation of the AP-1 complex, which normally only occur during the differentiation process of post-mitotic keratinocytes. Pharmacological impairment of this precocious AP-1 activation reduced the rate of apoptosis induced by CD437. The potent, selective, and p53 independent apoptosis-inducing efficacy of CD437 is of utmost importance for the prophylaxis and treatment of skin cancer caused by mutational inactivation of the p53 gene. PMID- 11862325 TI - Decoy oligonucleotide characterization of GATA-4 transcription factor in hypertrophic agonist induced responses of cardiac myocytes. AB - GATA-4 transcription factor is required for normal cardiac development. However, it is unknown whether GATA-4 is an essential mediator of hypertrophic responses in the heart. Rat B-type natriuretic peptide (BNP) gene promoter contains a region of two adjacent GATA binding sites (between -68 and -97) with high affinity for GATA-4. In order to block GATA-4 dependent signaling in cultured neonatal rat ventricular myocytes we administered a synthetic 30-bp phosphorothioated double-stranded DNA complementary to the rat BNP promoter region (between -68 and -97) as a "decoy" cis-element to bind GATA-4. GATA decoy oligodeoxynucleotide treatment of cardiomyocytes blocked GATA-4 DNA binding activity in electrophoretic mobility shift analysis and decreased baseline expression of cardiac natriuretic peptides and GATA-dependent promoter activity. In contrast, blocked GATA-4 DNA binding did not prevent endothelin-1 or phenylephrine induced expression of cardiac natriuretic peptides. Mutation of GATA binding sites at -80 and -91 rat BNP promoter downregulated baseline but did not affect endothelin-1 or angiotensin II induced promoter activity. Additively, GATA decoy oligodeoxynucleotide treatment was insufficient to block endothelin-1 induced activation of protein synthesis or sarcomeric protein assembly. In conclusion, a targeted disruption of GATA-4 DNA binding activity is insufficient to prevent hypertrophic agonist induced responses of ventricular myocytes. PMID- 11862327 TI - Amitriptyline and clomipramine activate Gi-protein signaling pathway in the induction of analgesia. AB - The post-receptorial mechanisms of the analgesic action of amitriptyline and clomipramine, two tricyclic antidepressants, were investigated in the mouse hot plate test by using an antisense strategy. Mice were injected i.c.v. with antisense oligonucleotides (aODN), complementary to the sequence of the mRNA sequence of the alpha-subunit of Gi1, Gi2 and Gi3-proteins, 18-24 h prior to the hot plate test. Treatment with aODN against Gi1alpha, Gi2alpha and Gi3alpha dose dependently reduced the analgesia induced by both amitriptyline (15 mg/kg s.c.) and clomipramine (25 mg/kg s.c.). This antagonistic effect disappeared 7 days after the end of the i.c.v. treatment, indicating the absence of irreversible damage or toxicity. Treatment with aODN against Gi1alpha, Gi2alpha and Gi3alpha, at the active doses, did not modify the animals' pain threshold, indicating the absence of any hyperalgesic effect. Amitriptyline, clomipramine and the aODN employed, at the maximal effective doses, did not produce any alteration of motor coordination of the mice, as revealed by rotarod experiments, and spontaneous motility, as revealed by the Animex apparatus. These results indicate that amitriptyline and clomipramine induce their analgesic effect by activating all three subtypes of the Gi-proteins. PMID- 11862328 TI - Biased inhibition by a suramin analogue of A1-adenosine receptor/G protein coupling in fused receptor/G protein tandems: the A1-adenosine receptor is predominantly coupled to Goalpha in human brain. AB - The interface between receptors and G proteins can be considered as a drug target. Various classes of low molecular weight inhibitors have been identified that block the ability of receptors to interact with G proteins (e.g. peptides, suramin analogues and amphiphilic cations). Here we have tested if there are compounds that differentially affect the interaction of one receptor with two different (related) G protein alpha-subunits. Fusion proteins comprising the human A1-adenosine receptor and Galphai-1 (A1/Galphai-1) or Galphao (A1/Galphao) were expressed in HEK293 cells. Suramin analogues were screened for their ability to differentially affect high affinity binding of the agonist (-) N6-3 [125I](iodo-4-hydroxyphenylisopropyl) adenosine (IHPIA). One compound [NF326 = 8,8'-(carbonylbis-(imino-3,1-phenylenecarbonylimino))bis-(1-naphthol-3,6 disulfonic acid, disodium salt)] was identified that inhibited high affinity agonist binding to the fusion protein A1/Galphai-1 but modestly enhanced binding of IHPIA to A1/Galphao. This action was specific because NF326 did not affect antagonist binding to either fusion protein. In addition, it was unrelated to a difference in affinity of the receptor for the G protein fusion moiety because the stability of ternary complexes formed by IHPIA + A1/Galphai-1 and IHPIA + A1/Galphao) is comparable and because lowering the affinity of the receptor for the G protein (by introducing point mutations at cys351 of Galphai-1) enhanced the uncoupling effect of NF326. Finally, NF326 did not discriminate between a fusion protein comprising the alpha2A-adrenoceptor and Galphai-1 (alpha2A)/Galphai-1) or Galphao-1 (alpha2A)/Galphao-1); binding of the agonist [3H]UK14304 (bromoxidine) to both fusion proteins was inhibited over a comparable concentration range while binding of the antagonist [3H]yohimbine was unaffected. These observations are consistent with the interpretation that the contact sites that are formed between individual receptors and G proteins differ. These differences suffice to allow for selective disruption by G protein inhibitors of different classes. Using NF326 we show that the bulk of the A1-adenosine receptors in human cerebrocortical membranes interacts with Galphao rather than Galphai. PMID- 11862329 TI - Inhibition of gastric emptying and small intestinal transit by ethanol is mediated by capsaicin-sensitive afferent nerves. AB - The neural mechanisms involved in mediation of the inhibitory effects of ethanol on the gastric emptying and the small intestinal transit were studied in adult male rats. The gastrointestinal transit was determined by measuring the amount of phenol red recovered from the stomach and small intestine after intragastric administration. Spinal and/or vagal peptidergic afferent nerves were subjected to selective denervation by chemodenervation techniques using capsaicin, a potent and specific sensory neurotoxin. Intragastric administration of a 2.5 g/kg body weight dose of ethanol resulted in inhibition of the gastric emptying and the small intestinal transit. Prior systemic treatment with capsaicin, which defunctionalizes both spinal and vagal capsaicin-sensitive afferent nerves, abolished the inhibitory effect of ethanol on the gastrointestinal transit. Similarly, selective chemodenervation of the vagal afferents by perineural capsaicin treatment completely blocked the effect of ethanol. These observations furnish evidence indicative of the involvement of capsaicin-sensitive neural pathways, and in particular vagal afferent nerves, in the mediation of the inhibitory effect of large doses of ethanol on the gastrointestinal motility. PMID- 11862330 TI - Effect of repeated treatment with high doses of selegiline on behaviour, striatal dopaminergic transmission and tyrosine hydroxylase mRNA levels. AB - The anti-parkinsonian drug selegiline is a monoamine oxidase B (MAO-B) inhibitor and a potential neuroprotective agent which facilitates dopaminergic transmission. Its metabolites (-)-amphetamine and (-)-metamphetamine might contribute to the pharmacological effects as they are also able to increase dopaminergic transmission and in addition might lead to behavioural sensitization after repeated administration. We investigated the effects of acute and repeated treatment with a high dose of selegiline on dopamine overflow in the striatum as well as on behaviour and on tyrosine hydroxylase (TH) mRNA levels in midbrain. Two experiments were performed. In the first one, rats were implanted with microdialysis probes into the striatum and received daily injections of selegiline (10 mg/kg, i.p.) for 1 or 8 days or a single dose of saline. In vivo microdialysis was carried out on days 1, 8 or 17 (after withdrawal of 9 days) to measure dopamine overflow. Motility was measured at the same time. In the second experiment, rats were injected daily with selegiline (10 mg/kg, i.p.) or saline over a time period of 6 weeks or only once before the brains were processed for in situ hybridization with a (35)S-radiolabelled probe for TH. Repeated treatment led to higher levels in motility scores than acute administration after administration of the same dose, indicating behavioural sensitization, which was still manifest after an interruption of 9 days in the supply of selegiline. In contrast, acute administration of selegiline increased dopamine levels to a similar degree as the same dose after subchronic treatment, with or without interruption of 9 days. The dopamine metabolite DOPAC was reduced by more than 50% after acute administration of selegiline and even more so on day 8 by the same dose, after repeated administration. The basal concentrations of dopamine (before challenge with selegiline) were not altered by the repeated administration, whereas the basal concentrations of DOPAC were decreased by more than 80% by the repeated administration of selegiline, suggesting a decrease in dopamine turnover. Acute administration did not have any influence on TH mRNA levels, whereas chronic treatment significantly reduced TH mRNA levels in substantia nigra and ventral tegmental area. In conclusion, repeated administration of selegiline leads to behavioural sensitization independent of altered dopamine levels. In addition, it leads to a decrease, probably due to a down-regulation, of dopamine turnover and tyrosine hydroxylase. PMID- 11862331 TI - Inhibition of cardiac potassium currents by pentobarbital. AB - The inhibitory effects of the anesthetic barbiturate pentobarbital on the slow ( I(Ks)) and fast component ( I(Kr)) of cardiac delayed rectifier potassium currents ( I(K)) and on the inward rectifier potassium currents ( I(K1)) were examined in Xenopus oocytes expressing the human minK, human ether-a-go-go related gene (HERG) and guinea pig Kir2.2, respectively. Block of native I(K) ( I(Ks) and I(Kr)) and inward rectifier potassium current ( I(K1)) by pentobarbital was examined in guinea pig ventricular myocytes. In oocytes using the two electrode voltage clamp technique potassium currents of hminK-, HERG- and Kir2.2 expressing oocytes were inhibited by pentobarbital with IC50 values of 0.20, 1.58 and 0.54 mM, respectively. I(Ks) block was time- and voltage-independent and had no influence on activation at positive voltages although it shifted voltage dependent activation to more positive voltages. Pentobarbital-induced HERG inhibition was not dependent on voltage but influenced the deactivation kinetics and shifted half-maximal activation to more negative voltages. In guinea pig cardiomyocytes, using the patch clamp technique, I(Ks) and I(Kr) were inhibited by pentobarbital with IC50 values of 0.18 mM and 2.75 mM, respectively. I(Kr) deactivation and I(Ks) activation kinetics were only slightly influenced by pentobarbital, if at all. Block of I(K1) was weakly voltage-dependent with IC(50) values of 0.26 mM (-40 mV) and 0.91 mM (-120 mV). The data show that pentobarbital suppresses both cloned ( I(K), I(Kir2.2)) and native ( I(K), I(K1)) cardiac potassium currents with the highest affinity for I(Ks). PMID- 11862332 TI - Kinetic model for the fate of 6-[18F]fluorodopamine in the human heart: a novel means to examine cardiac sympathetic neuronal function. AB - After injection of 6-[18F]fluorodopamine, thoracic positron emission tomographic scanning visualizes the sympathetic innervation of the heart. This report introduces a kinetic model that relates 6-[18F]fluorodopamine positron emission tomographic scanning results to specific aspects of cardiac sympathoneural function. Inputs were the 6-[18F]fluorodopamine concentration in arterial blood and the estimated contribution of circulating metabolites of 6 [18F]fluorodopamine. All of the three compartments in the model were intraneuronal. Two compartments corresponded to vesicles in sympathetic nerves, consistent with the "multiple vesicular pool" hypothesis from preclinical studies. The model successfully fit the empirical time-activity curve for myocardial 6-[18F]fluorodopamine-derived radioactivity and predicted correctly the effects of several neuropharmacological and physiological manipulations on the time-activity curve. Myocardial cell uptake of metabolites of 6 [18F]fluorodopamine from the circulation could explain an immediate peak of 6 [18F]fluorodopamine-derived radioactivity. The model seems useful in predicting effects of altered cardiac sympathetic function on time-activity curves for myocardial 6-[18F]fluorodopamine-derived radioactivity in humans. PMID- 11862333 TI - Spontaneous release of GDP from Gi proteins and inhibition of adenylyl cyclase in cardiac sarcolemmal membranes. AB - Several studies have shown an activation of adenylyl cyclase by the G protein inactivating guanine nucleotides, GDP and its phosphate transfer-resistant analog, guanosine 5'- O-(2-thiodiphosphate; GDPbetaS). Here, we studied the mechanism underlying this unconventional activation. Adenylyl cyclase activity in sarcolemmal membranes from failing human ventricular myocardium, at a low Mg2+ concentration, decreased rapidly during incubation at 37 degrees C. This decrease in enzyme activity was paralleled by a rapid release of GDP from Gi proteins, amounting to 75% release of total Gi-bound GDP within 10 min at 37 degrees C. In contrast, no GDP release was observed at 4 degrees C, and adenylyl cyclase activity remained stable for up to 20 min at 4 degrees C. GDPbetaS did not alter the initial rates of cyclic AMP formation by the adenylyl cyclase, at either 37 degrees C or 4 degrees C, but almost fully prevented the decrease in enzyme activity occurring at 37 degrees C. Hence, after 10 min of incubation at 37 degrees C, adenylyl cyclase activity in the presence of GDPbetaS was increased by 150%, while no difference in activity was observed at 4 degrees C. Under conditions where adenylyl cyclase is uncoupled from regulation by the inhibitory Gi proteins, i.e., at high concentrations of Mg2+ or Mn2+, adenylyl cyclase activity remained stable even at 37 degrees C and GDPbetaS did not stimulate activity. In conclusion, Gi proteins in sarcolemmal membranes from failing human hearts rapidly release bound GDP. The data, furthermore, suggest that this process results in adenylyl cyclase inhibition by the empty but apparently active Gi proteins. PMID- 11862334 TI - Evidence for the possible involvement of Ca2+ entry blockade in the relaxation by class I antiarrhythmic drugs in the isolated pig coronary smooth muscle. AB - Effects of several Na+ channel blockers (i.e., class I antiarrhythmic agents), procainamide, quinidine, lidocaine, mexiletine, propafenone, were investigated in the isolated endothelium-denuded pig coronary artery focusing on the possible involvement of the blockade of Ca2+ channels and/or opening of K+ channels in the relaxant responses. All drugs except procainamide induced a concentration dependent full relaxation of the coronary artery precontracted with high-KCl (30 mM, 80 mM). Inhibitions by procainamide of both high-KCl-induced contractions were less than 50% even at a concentration of 3 x 10(-2) M. Both high-KCl contractions were diminished by an L-type Ca2+ channel blocker, diltiazem, in a concentration-dependent manner. In contrast, cromakalim failed to inhibit 80 mM KCl-induced contraction. Tetrodotoxin (3 x 10(-5) M) did not affect the relaxant actions of the tested class I antiarrhythmic agents in high-KCl (80 mM)- or prostaglandin F2alpha-contracted muscle. The inhibitions by these class I antiarrhythmic agents of high-KCl-induced contraction were significantly attenuated when extracellular CaCl2 was increased from 2 mM to 20 mM. Furthermore, procainamide, quinidine, lidocaine, mexiletine as well as diltiazem decreased both cytoplasmic Ca2+ level ([Ca2+](cyt)) and muscle tension elevated by high-KCl in fura-2-loaded coronary preparations. These findings suggest that blockade of voltage-gated Ca2+ channels is involved in the relaxing action of these class I antiarrhythmic drugs in pig coronary artery. Blockade of Na+ channel and/or opening of K+ channels does not seem to play the principal role in the mechanism by which these antiarrhythmic drugs relax coronary artery. PMID- 11862335 TI - Human C-peptide acutely lowers glomerular hyperfiltration and proteinuria in diabetic rats: a dose-response study. AB - Recent studies suggested that C-peptide treatment of C-peptide-deficient patients with type I diabetes mellitus may present a new approach to prevent diabetic nephropathy. The present study further elucidated this concept by assessing the acute effect of human C-peptide application on kidney function in anesthetized rats with streptozotocin (STZ)-induced diabetes. Human C-peptide was applied as an i.v. bolus followed by continuous infusion of a fivefold dose per hour. A dose of 6 nmol/kg plus 30 nmol/kg per h is referred to as 1x. Application of 0.1, 0.3, 1, 3 or 10x to STZ-diabetic rats elicited mean plasma human C-peptide concentrations of 0.5, 5, 24, 75 and 225 nmol/l, respectively. Under basal conditions STZ-diabetic rats exhibited as expected an increase in glomerular filtration rate (GFR) by about 30%, which was associated with a lower total renal vascular resistance (RVR) and a rise in renal blood flow (RBF) as well as enhanced urinary protein excretion (UPE) of about 70% as compared with control rats. Human C-peptide dose-dependently lowered GFR and UPE in STZ-diabetic rats without altering blood glucose levels. No significant effect of human C-peptide on RBF or RVR could be detected, which may indicate an effect on glomerular ultrafiltration coefficient. Maximum effects of human C-peptide on the diabetes induced rises in GFR and UPE established an inhibition of 40% and 50%, respectively. Half-maximum effects on GFR and UPE were observed at plasma concentrations of human C-peptide in the range of 0.5-5 nmol/l, which is relatively close to endogenous C-peptide levels in non-diabetic rats. Unresponsiveness of non-diabetic control rats to human C-peptide further indicated specific effects. PMID- 11862336 TI - Okadaic acid, a protein phosphatase inhibitor, stimulates the activity of Na+/H+ and Na+-independent Cl-/HCO3- exchangers in human lymphocytes. AB - The cytosolic pH (pHi) regulatory mechanisms of peripheral blood human lymphocytes and the effect of okadaic acid on the activity of these mechanisms were studied by means of fluorescence imaging microscopy of 2',7'-bis (carboxyethyl)-5(6')-carboxyfluorescein (BCECF)-loaded individual cells. Lymphocytes recover from a CO(2)-induced acid load in an extracellular Na+ dependent, intracellular Cl- -independent fashion. This pHi recovery is highly sensitive to the anion exchange inhibitor 4,4'-diisothiocyanato-stilbene-2,2' disulfonic acid (DIDS) and minimally sensitive to the Na+/H+ exchange inhibitor amiloride, suggesting that it is mostly due to the action of a Na+-dependent HCO3 transporter. Extracellular Cl- and Na+ removal experiments point to the existence of a DIDS-sensitive Na+-independent Cl-/HCO3- exchanger. Preincubation with okadaic acid stimulates the pHi recovery rate from a CO2-induced acid load in the presence of DIDS (0.002 pHu/min vs. 0.065 pHu/min), but not in the presence of amiloride. Okadaic acid also accelerates the pHi elevation induced by Cl- removal (0.039 pHu/min vs. 0.067 pHu/min). In summary, these results indicate that okadaic acid stimulates the activity of Na+/H+ and Na+-independent Cl-/HCO3- exchangers, but has no effect on the activity of the Na+-dependent HCO3- transporter of human lymphocytes. PMID- 11862337 TI - Atypical kinetics for a series of putative dopamine antagonists to reverse the low-magnitude Ca2+ phase in the dopamine-bound D2short receptor state. AB - Real-time analysis of dopamine:antagonist interactions at the recombinant D2short receptor was performed by measuring time-dependent Ca2+ responses following activation of a chimeric Galphaq/o protein in CHO-K1 cells. Terguride (57%), +-UH 232 (20%) and buspirone (16%) demonstrated dopamine-like intrinsic activity at the presumably unoccupied, dopamine-free receptor; remoxipride, pipamperone and L 741626 being silent at 1 microM. Each of the putative antagonists (1 microM) displayed a transient reversal capacity of the low-magnitude Ca2+ phase in the dopamine-bound receptor state ( E(rev): 68%-92% vs. 1 microM tropapride) with a t1/2 between 8.8 s and 13.9 s upon antagonist addition; this capacity was either almost fully [remoxipride, pipamperone and +-UH 232] or partially [buspirone (31%), terguride (45%) and L 741626 (70%)] lost upon further incubation. The biphasic reversal Ca2+ profile of these dopamine antagonists is different from previously characterised dopamine antagonists which display either full reversal of the low-magnitude Ca2+ response with a fast or slow onset of action, or partial reversal stably present over the entire incubation period. The dynamic Ca2+ data strongly suggest that the dopamine D2short receptor can be blocked via multiple molecular mechanisms. PMID- 11862338 TI - Do we need placebos to evaluate new drugs in children with schizophrenia? AB - RATIONALE: Ethical issues are raised when placebos are used in human experimentation. This ethical concern is accentuated when research is contemplated among special sub-groups such as children, especially if they suffer from schizophrenia. OBJECTIVES: To review critically current ethical and scientific issues regarding the use of placebos in children with schizophrenia. METHODS: Manual review of literature and agency policies on pertinent issues and presentation of thoughtful original ideas as to the ethical use of placebos in children. RESULTS: There is compelling scientific and ethical justification underscoring the need and appropriateness of placebo-controlled studies in the treatment of childhood schizophrenia. CONCLUSION: The authors believe that children will gain access to new medications and receive safe and optimal drug therapy through well-conducted research that employs placebo controls. PMID- 11862339 TI - Reactivation of morphine conditioned place preference by drug priming: role of environmental cues and sensitization. AB - RATIONALE: Relapse is a major characteristic of drug addiction and remains the primary problem in treating drug abuse. Despite a great deal of research, the exact factors that determine renewed drug-seeking and persistent craving for them remain unclear. OBJECTIVE: The present study was designed to evaluate the role of environmental cues and behavioral sensitization in reactivation of place preference following long-term extinction of morphine conditioned place preference (CPP) in rats. METHODS: After being injected with morphine and saline alternately for 6 days to induce morphine CPP, the rats were subjected to extinction of conditioning for 21 days. The rats were then administered various doses of morphine, heroin, or cocaine and confined in the previous drug- or saline-paired compartment. CPP was determined. Some rats were treated with scopolamine or naloxone prior to administration of these three drugs. RESULTS: Morphine CPP disappeared following a 21-day extinction. A single injection of morphine, heroin, or cocaine evoked place preference for the previous drug-paired side. However, place preference for the previous vehicle-paired side was induced after the animals received a single injection of morphine, heroin or cocaine and confined to the previous vehicle-paired compartment. Administration of naloxone prior to drug treatment significantly attenuated the place preference induced by morphine or heroin, but had no significant effect on the place preference elicited by cocaine. Administration of the cholinergic antagonist scopolamine before morphine, heroin and cocaine inhibited the expression of place preference. CONCLUSIONS: Environment-related cues and behavioral sensitization play critical roles in the incentive motivation underlying drug-seeking behaviors. PMID- 11862340 TI - Histamine H3-receptor blockade in the rat nucleus basalis magnocellularis improves place recognition memory. AB - RATIONALE: Several lines of evidence have indicated that the central histaminergic system might be involved in learning and memory OBJECTIVES: The aim of the present study was to ascertain the impact on memory processes of putative histaminergic-cholinergic interactions in the nucleus basalis magnocellularis (NBM) of the rat. METHODS: The effects of thioperamide, a histamine H3-receptor antagonist, were studied on the memory performance of rats in a two-trial, delayed, place-recognition task. The drug was injected into the NBM area 2 min prior to the first trial (1.5, 7.5, and 37.5 ng/0.5 microl; pre-acquisition treatment), within 30 s from the end of the first trial (0.3, 1.5, 7.5, and 37.5 ng/0.5 microl; post-acquisition treatment), or 2 min prior to the second trial (1.5, 7.5, and 37.5 ng/0.5 microl; pre-retrieval treatment). RESULTS: Post acquisition intra-NBM injections of 1.5 ng and 7.5 ng, but not of 0.3 ng and 37.5 ng thioperamide, significantly enhanced memory retention in treated rats. The histamine H(3)-receptor blocker exerted pro-cognitive effects only when administered post-acquisition, since both pre-acquisition and pre-retrieval treatments were ineffective. The post-acquisition effect of the drug was time dependent and disappeared when the drug was injected 90 min after the end of the first trial. The U-shaped dose-response relationship and the time dependency of the effect of thioperamide indicated that the drug acts on mechanisms involved in memory consolidation. CONCLUSIONS: The present findings demonstrate that the pro cognitive effect of thioperamide is probably due to the modulation of post acquisition memory processes through an action on the cholinergic basal forebrain. Our results indicate also that H3-antagonists may provide a useful approach for improving spatial recognition memory. PMID- 11862341 TI - Acute effects of clomipramine and fluoxetine on dorsal periaqueductal grey-evoked unconditioned defensive behaviours of the rat. AB - RATIONALE: Several antidepressants attenuate conditioned escape behaviours reinforced by the terminus of an electrical stimulus applied to the dorsal periaqueductal grey (DPAG). OBJECTIVE: The present study examined whether the antidepressant and antipanic drugs clomipramine (CLM) and fluoxetine (FLX) also attenuate the DPAG-evoked unconditioned defensive behaviours. METHODS: Rats with electrodes in the DPAG were electrically stimulated in the absence of any treatment or 30 min after injections of CLM, FLX or saline. Threshold functions of cumulative response frequencies were fitted through the logistic model and compared using likelihood ratio coincidence tests. RESULTS: CLM produced non linear effects on galloping, for which median thresholds (I50) were significantly increased (19 +/- 2%) or decreased (22+/-2%) with 5 mg/kg and 10 mg/kg, respectively, or did not change with 20 mg/kg. The latter dose further increased the I(50) of micturition (38 +/- 1%) and decreased the defecation output (-33 +/- 15%). FLX significantly increased the I50 of immobility (22 +/- 2%) and galloping (25 +/- 3%) with 1 mg/kg and 5 mg/kg, respectively. Moreover, corresponding doses either decreased the maximum output (-25 +/- 13%) or increased the I50 (56 +/- 11%) of defecation. Saline was ineffective. CONCLUSIONS: While the attenuation of defecation and micturition by 20 mg/kg CLM suggests a peripheral antimuscarinic action, CLM non-linear effects on galloping were most likely due to its differential action on monoaminergic and cholinergic central mechanisms. In contrast, the attenuation of immobility, galloping and defecation by low doses of FLX suggests a serotonin-mediated antiaversive action. Finally, CLM and FLX acute effects on DPAG-evoked unconditioned galloping response were strikingly similar to those reported for DPAG-evoked shuttle-box conditioned escape. PMID- 11862342 TI - Comparison of interactions of D1-like agonists, SKF 81297, SKF 82958 and A-77636, with cocaine: locomotor activity and drug discrimination studies in rodents. AB - RATIONALE: Recent data suggest that dopamine (DA) D1-like receptor full agonists may be potential pharmacotherapeutic agents for treating cocaine abuse. The structurally novel isochroman D1-like agonist, A-77636, has not been well characterized and may prove to be useful as such an agent. OBJECTIVES: The interactions of cocaine and A-77636 were compared to those obtained with the better investigated benzazepine D1-like dopamine agonists, SKF 82958 and SKF 81297. The alterations in the locomotor stimulant and discriminative-stimulus effects of cocaine by the full D1-like dopamine receptor agonists were investigated across a full range of doses in order to characterize their interactions. METHODS: Drug-naive Swiss-Webster mice were pretreated with SKF 81297, SKF 82958 or A-77636 (1-10 mg/kg) and cocaine (5-56 mg/kg) prior to a 30 min period in which locomotor activity was assessed. Rats were trained on a fixed ratio 20 (FR20) schedule to discriminate IP saline from cocaine (10 mg/kg) injections. Cocaine alone (1-10 mg/kg) and with either A-77636 (0.56-1.7 mg/kg), SKF 82958 (0.01-0.1 mg/kg) or SKF 81297 (0.1-0.56) were injected IP 5 min prior to a 15-min test session. RESULTS: Cocaine maximally stimulated activity at 20-40 mg/kg with higher and lower doses stimulating activity less. Each D1-like agonist produced a dose-related decrease in cocaine-induced locomotor activity and lowered its maximal rate. Each of the D1-like agonists partially substituted for cocaine, with maximal substitution approximating 49, 35, and 24% for SKF 81297, SKF 82958, and A-77636, respectively. SKF 82958 significantly shifted the cocaine dose-effect curve approximately 3-fold to the left. With SKF 81297, there was a trend towards a leftward shift of cocaine dose effects, however the change was not statistically significant. In contrast to the other two D1-like agonists, A 77636 either did not affect the cocaine dose-effect curve or shifted it to the right. CONCLUSIONS: All three agonists produced similar effects on cocaine induced locomotor activity, however the discriminative-stimulus effects of cocaine were affected differently by the D1 agonists. These results suggest fundamental differences in the actions of these D1 agonists. Because A-77636 consistently attenuated the present effects of cocaine, it may prove more useful than the others as a pharmacotherapy to treat cocaine abuse. PMID- 11862343 TI - Cocaine-induced brain activation determined by positron emission tomography neuroimaging in conscious rhesus monkeys. AB - RATIONALE: Cerebral blood flow can provide a useful dependent measure to characterize cocaine-induced changes in brain function. The acute effects of cocaine administration on cerebral blood flow may have direct relevance to the etiology of cocaine addiction. OBJECTIVES: The present study used positron emission tomography (PET) neuroimaging techniques to characterize the acute effects of cocaine administration on cerebral blood flow in conscious rhesus monkeys. METHODS: Functional changes in cerebral blood flow were determined in four drug-naive subjects with the positron-emitting tracer 15O water following acute i.v. administration of cocaine (0.3 and 1.0 mg/kg). Specific attention was devoted to the development of an effective and comfortable head restraint device to use in the imaging of conscious monkeys. Experimental sessions comprised eight consecutive i.v. injections of 15O water at 10-min intervals. PET scans of 90 s duration occurred 10 s after each injection. RESULTS: Repeated baseline determinations of cerebral blood flow prior to drug administration were reliable. Cocaine had significant, dose-related effects on cerebral blood flow at 5 min postinjection that diminished relative to control (saline) conditions by 15 min postinjection. Brain activation maps normalized to global flow showed prominent cocaine-induced activation of prefrontal cortex localized primarily to dorsolateral regions. Importantly, cocaine-induced brain activation was blocked by pretreatment with the selective serotonin uptake inhibitor, alaproclate (3.0 and 10.0 mg/kg). The results document a distinct pattern of cocaine-induced brain activation that is sensitive to pharmacological manipulation. CONCLUSION: The pattern of brain activation induced by acute administration of cocaine may provide a useful means to evaluate medication effectiveness for treating cocaine addiction. PMID- 11862344 TI - Blockade of hydromorphone effects by buprenorphine/naloxone and buprenorphine. AB - RATIONALE: Buprenorphine is an opioid agonist-antagonist used in the treatment of opioid dependence. Naloxone has been combined with buprenorphine to decrease the parenteral abuse potential of buprenorphine. This addition of naloxone may also confer further opioid blockade efficacy. OBJECTIVES: To test the opioid blockade efficacy of sublingual buprenorphine/naloxone versus buprenorphine alone and determine whether: (1) the blockade efficacy of buprenorphine/naloxone varies between the time of expected maximal and minimal effects of naloxone, (2) the blockade efficacy of buprenorphine/naloxone and buprenorphine varies as a function of maintenance dose level, and (3) there are adaptive changes over time associated with repeated daily dosing of buprenorphine/naloxone and buprenorphine. METHODS: Residential subjects ( n=6) were maintained on different double-blind dose levels of buprenorphine/naloxone (4/1, 8/2, 16/4, 32/8 mg) and buprenorphine (32 mg) for 6-day periods and challenged with parenteral doses of hydromorphone (12 mg) in laboratory sessions. RESULTS: There was no evidence of additional opioid blockade efficacy conferred by combining naloxone with buprenorphine. Higher doses of buprenorphine/naloxone provided greater blockade of hydromorphone effects. Changes over time associated with repeated daily dosing of buprenorphine/naloxone and buprenorphine were minimal. CONCLUSIONS: The addition of naloxone to buprenorphine may deter the parenteral abuse of buprenorphine/naloxone, but it does not enhance the therapeutic efficacy of buprenorphine. The blockade efficacy of buprenorphine/naloxone is dose related; however, doses up to 32/8 mg buprenorphine/naloxone provide only partial blockade when subjects receive a high dose of an opioid agonist. PMID- 11862345 TI - A study of the effect of a single neurotoxic dose of 3,4 methylenedioxymethamphetamine (MDMA; "ecstasy") on the subsequent long-term behaviour of rats in the plus maze and open field. AB - RATIONALE: Decreased 5-HT function has been shown to induce behaviour consistent with an "anxiolytic" effect. Administration of a single dose of 3,4 methylenedioxymethamphetamine (MDMA; "ecstasy" 12.5 mg/kg IP) to rats results in prolonged damage to central serotonergic nerve terminals. Thus we wished to assess whether an MDMA-induced lesion may have longer-term behavioural consequences. OBJECTIVE: The study was designed to examine the behaviour of MDMA pretreated and control animals in the elevated plus-maze and open field at a number of time-points, up to 80 days, after the administration of a single neurotoxic dose of MDMA (12.5 mg/kg IP). RESULTS: MDMA-pretreated Dark Agouti rats demonstrated a statistically significant reduction in anxiety-related behaviour, compared to saline-pretreated control rats, in both the elevated plus maze and open field when the rats were tested on day 73 (open field) and day 80 (plus maze) after MDMA administration. CONCLUSIONS: The behavioural consequences of a single neurotoxic dose of MDMA can be demonstrated over 2 months after administration of the compound, thereby indicating that long-term adaptive changes occur within the brain following the neurodegeneration of 5-HT neurones produced by this recreationally used drug. PMID- 11862346 TI - The light-enhanced startle paradigm as a putative animal model for anxiety: effects of chlordiazepoxide, flesinoxan and fluvoxamine. AB - RATIONALE: Recently, a new putative animal model of anxiety, "light-enhanced startle" was introduced. By placing a rat in a brightly lit environment, which is a naturally aversive stimulus to rats, the amplitude of the startle response to a startle-eliciting noise burst is increased. OBJECTIVES: The present study aimed to determine the predictive validity of the light-enhanced startle as a putative model for anxiety. METHODS: The effects of the GABA(A)-benzodiazepine receptor agonist chlordiazepoxide (CDP), the 5-HT1A receptor agonist flesinoxan and the specific 5-HT reuptake inhibitor fluvoxamine on light-enhanced startle were studied. RESULTS: Both CDP and flesinoxan decreased startle potentiation, whereas fluvoxamine was devoid of any effects on potentiation. Effects on baseline startle amplitude were only seen after CDP administration. CONCLUSIONS: The present experiment provides evidence for the predictive validity of the light enhanced startle as an animal model for anxiety. Due to the use of an unconditioned anxiogenic stimulus, the light-enhanced startle offers several benefits over animal models that depend on conditioning. Drug effects can be ascribed more directly to effects on anxiety, as opposed to memory retrieval and, as shown in this study, non-specific drug effects can easily be detected without the interference of contextual fear. PMID- 11862347 TI - Stimulation of voluntary ethanol intake by cannabinoid receptor agonists in ethanol-preferring sP rats. AB - RATIONALE: Recent studies have shown that the cannabinoid CB1 receptor antagonist, SR 141716, is capable of reducing voluntary ethanol intake in rodents, suggesting the involvement of the CB1 receptor in the neural circuitry mediating the positive reinforcing properties of ethanol. OBJECTIVES: The present study extended to the agonists the investigation on the pharmacological manipulation of ethanol intake by cannabinoid agents. METHODS: Selectively bred, Sardinian alcohol-preferring (sP) rats were offered ethanol and water under the two-bottle free choice procedure with unlimited access for 24 h/day. RESULTS: The acute administration of WIN 55,212-2 (0.5-2 mg/kg; IP) and CP 55,940 (3-30 microg/kg; IP) induced a significant, dose-dependent increase in ethanol intake. Conversely, water consumption and intake of regular food and a highly palatable sucrose solution were not affected by treatment with WIN 55,212-2 and CP 55,940. The stimulatory effect of WIN 55,212-2 and CP 55,940 on ethanol intake was completely prevented by administration of SR 141716 (0.3 mg/kg; IP) and the opioid receptor antagonist, naloxone (0.1 mg/kg; IP). CONCLUSIONS: Administration of WIN 55,212-2 and CP 55,940 promoted voluntary ethanol intake in sP rats. This effect was mediated by stimulation of the cannabinoid CB1 receptor and required the activation of the endogenous opioid system. The results of the present study add further support to the hypothesis that the cannabinoid CB1 receptor is part of the neural substrate regulating ethanol intake. These results are also discussed in terms of WIN 55,212-2 and CP 55,940 administration possibly fixing to a higher level the hedonic set-point mechanism regulating ethanol drinking behavior in sP rats. PMID- 11862348 TI - A comparison between Dark Agouti and Sprague-Dawley rats in their behaviour on the elevated plus-maze, open-field apparatus and activity meters, and their response to diazepam. AB - RATIONALE: Preliminary unpublished studies in our laboratory suggested that the behaviour of Sprague-Dawley (SD) and Dark Agouti (DA) rats was markedly different on both the elevated plus maze and in the open-field apparatus. We wished to confirm and extend this initial finding. OBJECTIVE: The study was designed to examine the behaviour of SD and DA rats in the elevated plus maze, open-field apparatus and automated activity meters. The response of both strains on the elevated plus maze following diazepam (1 mg/kg and 1.5 mg/kg) administration was subsequently investigated. RESULTS: DA rats showed markedly greater anxiety-like behaviour than SD rats in both the plus maze and open field, with fewer percentage open/total arm entries and percentage time spent on open/total arms in the plus maze and fewer crossings in the open field. Acute handling plus administration of vehicle abolished this difference in anxiety levels, with DA rats showing similar open-arm behaviour to that of SD rats. Both strains demonstrated a clear anxiolytic response to diazepam (1 mg/kg) in terms of percentage time spent on the open arms, but only SD rats had a statistically significant increase in percentage open-arm entries compared with vehicle injected control animals. CONCLUSIONS: While the high level of anxiety-like behaviour of DA rats versus SD rats could prove useful in future ethological studies on anxiety, the fact that acute handling decreased the anxiety-like behaviour on the elevated plus maze may limit the value of this strain for the study of putative anxiolytic drugs. PMID- 11862349 TI - Repeated electroconvulsive shock attenuates the depressive-like effects of d amphetamine withdrawal on brain reward function in rats. AB - RATIONALE: The withdrawal of humans from high doses of psychostimulant drugs can result in a transient syndrome which appears isomorphic to endogenous depression. One of the more prominent symptoms is a loss of hedonic capacity; in animals, the anhedonia associated with amphetamine withdrawal has been measured objectively by decrements in responding for intracranial self-stimulation (ICSS). OBJECTIVE: To date, the effects of amphetamine withdrawal on ICSS responding have been reversed by different antidepressant drugs. In the present study, we sought to reverse withdrawal-induced anhedonia by administration of repeated electroconvulsive shocks (ECS). METHODS: Rats with electrodes in the lateral hypothalamus were trained on an ascending-series current intensity ICSS paradigm until stable levels of responding were attained. Half of the animals were then administered a 4-day escalating dose schedule of d-amphetamine, and tests for ICSS responding started 12 h after the final injection. During withdrawal, all animals received daily treatment with either ECS or sham-ECS. RESULTS: Amphetamine withdrawal was associated with reduced ICSS responding; animals treated with ECS exhibited a facilitated recovery compared to sham-ECS treated animals, and returned to control levels of ICSS responding 24 h earlier. CONCLUSIONS: ECS was able to mitigate the anhedonic effects of d-amphetamine withdrawal, and provides additional support for the use of psychostimulant withdrawal as a model of depression. PMID- 11862350 TI - A comparative study of the effects of carbamazepine and the NMDA receptor antagonist remacemide on road tracking and car-following performance in actual traffic. AB - RATIONALE: Antiepileptic drugs are known to produce side effects which may impair driving performance. Performance effects, however, may differ substantially between individual antiepileptic drugs. OBJECTIVE: To compare the effects of carbamazepine, remacemide, and placebo on actual driving performance during a 12 day incremental dosing regimen. METHODS: Twenty-two healthy volunteers participated in a three-way, double-blind, cross-over driving study. Treatment effects were assessed in two actual driving tests carried out on days 8, 10, and 12 of each treatment period. The Road Tracking Test involved driving an instrumented vehicle at a constant speed and steady lateral position between the delineated lane boundaries. Standard deviation of lateral position (SDLP) was measured to indicate precision of road tracking control. The Car-Following Test involved driving the same vehicle behind a leading car and maintaining that distance while the latter executed a series of deceleration/acceleration maneuvers. Time to speed adaptation (TSA) and brake reaction time were the primary measures. RESULTS: Remacemide did not affect the subjects' driving performance. Carbamazepine increased SDLP throughout treatment and lengthened TSA on day 8. Changes in SDLP relative to placebo were comparable to those previously seen in drivers conducting the same test with blood alcohol concentrations of 0.05 g/dl. CONCLUSION: Remacemide, at the given dose regimen, does not affect driving performance. Carbamazepine, at the given dose regimen, can produce mild but sufficient impairment to put epileptic patients at risk when driving, at least during initiation therapy. PMID- 11862351 TI - The impact of tricyclic antidepressants and selective serotonin re-uptake inhibitors on handwriting movements of patients with depression. AB - RATIONALE: Psychomotor retardation is a common symptom of patients with major depressive disorder. While a variety of clinical examinations using different techniques have been undertaken to assess the motor component of psychomotor retardation in depression, the effects of antidepressants on psychomotor functions have been examined less extensively. OBJECTIVE: The aim of the present study was to examine the effect of various pharmacological treatments on handwriting movements of patients with depression. METHODS: Kinematic data of automated handwriting movements of 18 depressed patients receiving tricyclic antidepressants (TCAs), 18 patients on selective serotonin re-uptake inhibitors (SSRIs) and 18 healthy subjects was recorded and analysed. Groups were matched according to age, sex, handedness and education level. For the assessment of fine motor movements, a digitising tablet was used. Subjects were asked to perform a simple writing task. Movement time, velocity and acceleration of the handwriting movements were measured. RESULTS: Statistical analysis of writing movements revealed motor slowing in patients receiving TCAs. In comparison with both healthy subjects and patients receiving SSRIs, the TCA group displayed an increased movement time, reduced automation of handwriting, lower maximum velocities and reduced acceleration of descending strokes. CONCLUSIONS: The results suggest either that TCAs have adverse effects on motor functioning or that they are less effective in the treatment of motor retardation than SSRIs. PMID- 11862352 TI - Difference in serotonergic and noradrenergic regulation of human social behaviours. AB - RATIONALE: Treatment with antidepressants has been shown to affect social functioning, but drugs with actions on different neurotransmitters may have a different profile of effects. OBJECTIVE: To study the effects of acute manipulation of two neurotransmitters, serotonin and noradrenaline, on social behaviour in healthy volunteers. METHODS: Sixty volunteers were randomly assigned to a single dose of a selective noradrenaline reuptake inhibitor, reboxetine (4 mg), a selective serotonin reuptake inhibitor, citalopram (10 mg), or placebo. They socially interacted with a confederate behaving in a non-sociable manner in a stranger-dyadic social interaction paradigm 1.5 h postdrug. Social behaviour during the interaction was video recorded by a hidden camera and subsequently analysed. After the interaction, volunteers played the mixed-motive game with the confederate. This game has been shown to measure cooperative behaviour and communication. Volunteers read a short story and rated their mood predrug and before and after the interaction. RESULTS: Subjects on reboxetine showed reduced hand fiddling during the interaction and gave significantly more cooperative communications during the mixed-motive game. More volunteers on reboxetine were classified as cooperative players. On the reading task, the speech of subjects on citalopram showed less reduction of energy variation after the social interaction. CONCLUSION: Reboxetine had clear effects on social behaviour. Noradrenaline was related to increased social engagement and cooperation and a reduction in self-focus. Citalopram had less effect on cooperative behaviour but serotonin may be associated with protection of the self from the negative consequences of social interaction. PMID- 11862353 TI - Effect of branch chain amino acids supplemented with tryptophan on tyrosine availability and plasma prolactin. PMID- 11862355 TI - Interactions between dopamine D1 receptors and gamma-aminobutyric acid mechanisms in substantia nigra pars reticulata of the rat: neurochemical and behavioral studies. AB - RATIONALE: Several studies have shown that dopamine D1 agonists act on forebrain dopamine terminal regions to exert many of their behavioral effects. Yet, there is also a large number of D1 receptors in the substantia nigra pars reticulata (SNr), and these receptors are located mainly on terminals of gamma-aminobutyric acid (GABA)-ergic striatonigral neurons. OBJECTIVE: The present studies were undertaken to determine the behavioral and neurochemical effects of local administration of the D1 agonist SKF 82958 and to study the interactions between D1 and GABA mechanisms in SNr. METHODS: Microdialysis methods were used to characterize the effect of SKF 82958 on extracellular GABA, and several experiments studied the effects of nigral D1 stimulation on motor activity and investigated the behavioral significance of D1/GABA interactions in SNr. RESULTS: Local infusion of 10(-6) M SKF 82958 increased extracellular levels of SNr GABA, and this effect was blocked by co-infusion of the D1 antagonist SCH 23390. Bilateral SNr injections of SKF 82958 increased locomotor activity, and this effect was blocked by the GABA-A antagonist bicuculline. Intranigral bicuculline reduced motor activity, while the GABA-A agonist muscimol increased various motor activities in a manner similar to SKF 82958. CONCLUSIONS: The present results suggest that the D1 agonist SKF 82958 acts on D1 receptors in SNr to increase extracellular levels of GABA, and the increase in motor activity produced by nigral D1 stimulation is dependent on stimulation of GABA-A receptors. D1/GABA interactions in SNr are important for the modulation of basal ganglia output, which may have important implications for Parkinson's disease. PMID- 11862356 TI - Maintaining alertness and performance during sleep deprivation: modafinil versus caffeine. AB - RATIONALE: The performance and alertness effects of modafinil were evaluated to determine whether modafinil should replace caffeine for restoring performance and alertness during total sleep deprivation in otherwise healthy adults. OBJECTIVES: Study objectives were to determine (a) the relative efficacy of three doses of modafinil versus an active control dose of caffeine 600 mg; (b) whether modafinil effects are dose-dependent; and (c) the extent to which both agents maintain performance and alertness during the circadian trough. METHODS: Fifty healthy young adults remained awake for 54.5 h (from 6:30 a.m. day 1 to 1:00 p.m. on day 3) and performance and alertness tests were administered bi-hourly from 8:00 a.m. day 1 until 10:00 p.m. day 2. At 11:55 p.m. on day 2 (after 41.5 h awake), subjects received double blind administration of one of five drug doses: placebo; modafinil 100, 200, or 400 mg; or caffeine 600 mg ( n=10 per group), followed by hourly testing from midnight through 12:00 p.m. on day 3. RESULTS: Performance and alertness were significantly improved by modafinil 200 and 400 mg relative to placebo, and effects were comparable to those obtained with caffeine 600 mg. Although a trend toward better performance at higher modafinil doses suggested a dose-dependent effect, differences between modafinil doses were not significant. Performance enhancing effects were especially salient during the circadian nadir (6:00 a.m. through 10:00 a.m.). Few instances of adverse subjective side effects (nausea, heart pounding) were reported. CONCLUSIONS: Like caffeine, modafinil maintained performance and alertness during the early morning hours, when the combined effects of sleep loss and the circadian trough of performance and alertness trough were manifest. Thus, equivalent performance- and alertness enhancing effects were obtained with drugs possessing different mechanisms of action. However, modafinil does not appear to offer advantages over caffeine (which is more readily available and less expensive) for improving performance and alertness during sleep loss in otherwise normal, healthy adults. PMID- 11862357 TI - Effects of alpha 4/beta 2- and alpha 7-nicotine acetylcholine receptor agonists on prepulse inhibition of the acoustic startle response in rats and mice. AB - RATIONALE: Nicotine and agonists at subtypes of the nicotine acetylcholine receptor (nAChR) affect auditory gating, but the magnitude and direction of such effects appear highly variable. This variability may be due to differences in the tested dose range, selectivity of the test compound, species and strain, and suggests that nAChR subtypes are differentially involved in the control of auditory gating. OBJECTIVES AND METHODS: This study aimed to characterise the effects of nicotine and agonists with preferential activity at alpha4/beta2- and alpha7-nAChRs on auditory sensorimotor gating using a prepulse inhibition (PPI) paradigm. Similar experimental conditions were employed in rats and two strains of mice. The paradigm used startle stimuli of 120 dB and prepulse intensities of 3, 6 and 12 dB above a background of 70 dB. RESULTS: In Sprague-Dawley rats, nicotine disrupted PPI [minimal effective dose (MED): 1 mg/kg, SC] and this effect was mimicked by the potent nAChR agonist, epibatidine, (MED: < or = 0.001 mg/kg, IP) and the potent, and relatively selective, alpha4/beta2-nAChR agonist A 85380 (MED: < or = 0.1 mg/kg, IP). The effects of epibatidine, A-85380 and, to a lesser extent, nicotine were blocked by the non-selective nAChR antagonist mecamylamine. The relatively selective alpha7-nAChR agonists, GTS-21 and AR-R 17779, did not affect PPI in a consistent manner, both in rats and in DBA/2 mice, a strain expressing a disrupted gating phenotype, presumably due to altered activity of hippocampal alpha7-nAChRs. In BALB/c mice, a strain expressing a normal gating phenotype, nicotine (MED: 10 mg/kg, SC), epibatidine (MED: 0.03 mg/kg, IP) and A-85380 (MED: 0.3 mg/kg, IP) predominantly augmented PPI and mecamylamine attenuated these effects. CONCLUSIONS: The present results confirm that the effects of nAChR agonists on PPI are species dependent and suggest that stimulation of heteromeric nAChRs containing both alpha and beta subunits, and possibly of the alpha4/beta2 type, affect sensorimotor gating. Evidence supporting a role for alpha7-nAChRs in the control of PPI of the acoustic startle response was not obtained. PMID- 11862358 TI - Environmental context conditioning with ethanol reduces the aversive effects of ethanol in the acquisition of self-administration in rats. AB - RATIONALE: Many recent theoretical approaches to drug-taking behavior feature a role for Pavlovian conditioning. Despite growing evidence for that role, the particular contributions of Pavlovian conditioning to self-administration are not clear. For example, few studies have addressed the effects of Pavlovian conditioning on the acquisition of self-administration. OBJECTIVES: The purpose of this study was to test the effect of Pavlovian conditioning with an environmental conditioned stimulus and an ethanol unconditioned stimulus on the acquisition of self-administration reinforced by ethanol. METHODS: Rats were either given ethanol by gastric gavage in a distinctive context or in their home cage. All animals were then trained to bar press on a variable interval schedule for a sweetened ethanol solution in the distinctive context. RESULTS: Animals that had received ethanol associated with the training context maintained a higher level of bar press behavior for ethanol as the reinforcing solution. This effect developed only after the first session and resulted from differences in response rates, but did not affect the rate of reinforcement. CONCLUSIONS: This study demonstrates that an environmental context signaling the effects of ethanol maintains a higher operant response rate when ethanol is used subsequently as a reinforcer. This finding replicates previous reports of Pavlovian conditioning effects on ethanol consumption. The specific pattern of results suggests that conditioned tolerance modifies the reinforcing impact of ethanol. Context conditioning with ethanol reduces the aversive impact of initial ethanol consumption and maintains the reinforcing value of the ethanol solution. PMID- 11862359 TI - Effects of chronic paroxetine administration on measures of aggressive and impulsive responses of adult males with a history of conduct disorder. AB - RATIONALE: The role of serotonin in human aggression and impulsivity was evaluated by administering paroxetine or placebo for 3 weeks and comparing the effects on laboratory measures of aggression and impulsivity among male subjects with a history of conduct disorder. METHODS: Twelve male subjects with a history of criminal behavior participated in experimental sessions, which measured aggressive and impulsive responses. Six subjects were assigned to placebo treatment and six subjects to placebo and paroxetine treatment. Aggression was measured using the point subtraction aggression paradigm (PSAP), which provides subjects with an aggressive and monetary reinforced response options. Impulsive responses were measured using a paradigm that gives subjects choices between small rewards after short delays versus larger rewards after longer delays. RESULTS: Chronic administration of paroxetine (20 mg/day) for 21 days produced significant decreases in impulsive responses. Decreases in aggressive responses were evident only at the end of paroxetine treatment. Decreases in impulsive and aggressive responses could not be attributed to a non-specific sedative action because monetary reinforced responses were not decreased as has been observed following CNS sedation. CONCLUSIONS: Inhibition of serotonin reuptake by paroxetine is the possible mechanism for reductions in aggressive and impulsive responses. These results support other data linking serotonin function and aggression and impulsivity. PMID- 11862360 TI - Effects of gonadal steroid hormone treatments on opioid antinociception in ovariectomized rhesus monkeys. AB - RATIONALE: Gonadal steroid hormones altered opioid antinociception under some conditions in rodents, and we reported previously that chronic estradiol enhanced kappa but not mu opioid antinociception in ovariectomized rhesus monkeys. Sex differences have also been observed in the antinociceptive effects of opioid agonists. These findings suggest that gonadal hormones may modulate opioid antinociception. OBJECTIVES: To extend our previous studies of estradiol by examining the effects of progesterone alone, estradiol in combination with progesterone, and testosterone alone on opioid antinociception in ovariectomized rhesus monkeys. METHODS: Opioid effects were studied during chronic treatment with vehicle (sesame oil) or with progesterone alone (P; 0.32 mg/kg per day), a combination of progesterone+estradiol (P+E; 0.32 mg/kg per day P + 0.002 mg/kg per day E), or testosterone alone (T; 0.32 mg/kg per day). Opioid antinociception in a warm-water tail-withdrawal procedure was examined with the selective kappa opioid agonist U50,488, the selective mu agonist morphine, and the two mixed action opioids butorphanol and nalbuphine. RESULTS: The steroid treatment regimens produced physiological levels of progesterone and estradiol similar to peak levels observed during the luteal phase of the menstrual cycle and physiological levels of testosterone similar to those observed in intact males. Treatment with P, P+E, or T did not alter baseline thermal nociception. P+E significantly increased the potency of U50,488 at 50 degrees C but not at 54 degrees C. Gonadal hormone treatments had little or no effect on antinociception produced by morphine, butorphanol, or nalbuphine at either temperature. CONCLUSIONS: These findings further suggest that chronic treatment with physiological levels of gonadal hormones may modulate the antinociceptive effects of U50,488 in ovariectomized rhesus monkeys. PMID- 11862361 TI - Blockade or stimulation of D1 dopamine receptors attenuates cue reinstatement of extinguished cocaine-seeking behavior in rats. AB - RATIONALE: D(1) dopamine receptor antagonists and agonists attenuate cocaine reinstatement of cocaine-seeking behavior (i.e., responding in the absence of cocaine reinforcement). OBJECTIVES: The present study investigated the effects of a D(1) antagonist (SCH-23390), partial agonist (SKF-38393), and full agonist (SKF 81297) on reinstatement of cocaine-seeking behavior elicited by presentation of cocaine-paired cues. METHODS: Rats that had been trained to self-administer cocaine with a light/tone stimulus complex paired with each infusion underwent extinction across days. After responding diminished, rats were given response contingent access to the cocaine-paired stimulus complex. The effects of SCH 23390 (0-10.0 microg/kg), SKF-38393 (0-3.0 mg/kg), and SKF-81297 (0-3.0 mg/kg) on cue reinstatement of cocaine-seeking behavior were examined. The ability of the two D(1) agonists to independently reinstate cocaine-seeking behavior and the effects of SKF-81297 on cocaine reinstatement were also examined. To investigate the possibility of behavioral interference, the effects of SKF-38393 and SKF 81297 on grooming and stereotypy were assessed. RESULTS: SCH-23390 and SKF-81297, but not SKF-38393, attenuated cue reinstatement. However, while SKF-81297 dose dependently increased response latency, SCH-23390 did not. SKF-81297 also independently reinstated responding at the two lowest doses tested while SKF 38393 had no effect. Furthermore, SKF-81297 decreased cocaine reinstatement and increased response latency only at the highest dose. Finally, stereotypy was observed at all doses of SKF-81297 that also decreased responding, although the patterns of changes in these behaviors did not completely correspond. CONCLUSIONS: While the antagonist and full agonist produced similar effects on cocaine-seeking behavior, only the agonist increased response latency, suggesting that different processes mediate the effects of these drugs. PMID- 11862362 TI - Ecstasy (MDMA): are the psychological problems associated with its use reversed by prolonged abstinence? AB - RATIONALE: Chronic, regular recreational use of ecstasy (MDMA) is associated with psychopathology, elevated behavioural impulsivity and persistent impairment of memory performance. OBJECTIVE: The aim of the present study was to investigate which of these sequelae persist after at least 6 months of abstinence from ecstasy. METHODS: Four groups of participants were compared: 18 current regular recreational ecstasy users, 15 ex-regular ecstasy users who had abstained from using the drug for an average of 2 years, 16 polydrug users who had never taken ecstasy and 15 drug-naive controls. RESULTS: There were no significant group differences in age, education level, or pre-morbid intelligence and, generally, the use of illicit drugs other than ecstasy was not significantly different among the three drug-using groups. Both current and ex-ecstasy users exhibited elevated psychopathology and behavioural impulsivity compared with polydrug users and drug naive controls, but current ecstasy users exhibited a broader range of psychopathology than ex-users. Both groups of ecstasy users also exhibited impaired working memory and verbal recall performance compared with drug-naive controls, but only ex-users exhibited impaired verbal recall relative to polydrug users. Regression analysis indicated that psychopathology was primarily predicted by the extent of previous consumption of cannabis rather than ecstasy, whereas the majority of the cognitive deficits were only predicted by the extent of previous ecstasy use. CONCLUSIONS: Selective impairments of neuropsychological performance associated with regular ecstasy use are not reversed by prolonged abstinence. This is consistent with evidence that ecstasy has potent and selective neurotoxic effects on brain serotonergic systems in humans. PMID- 11862363 TI - Light-enhanced startle: further pharmacological and behavioral characterization. AB - RATIONALE: Previous findings indicate that the acoustic startle response is elevated when rats are tested in bright light. The phenomenon is disrupted by the 5HT1A partial agonist and the D2 receptor antagonist buspirone, a compound that also blocks the effect on startle of conditioned fear, and it was suggested that light-enhanced startle reflects an anxious state produced by bright light. It was also suggested that pre-test handling may be necessary for light-enhanced startle. To characterize this phenomenon further, we evaluate here the sensitivity of light-enhanced startle to the anxiolytic compound chlordiazepoxide, to the noradrenergic beta-receptor antagonist propranolol, and to pre-test handling. METHODS: Startle was measured for 20 min in the dark (phase I), followed 5 min later by a second test (phase II) either in the dark or the light. Immediately prior to testing, rats received IP injections of chlordiazepoxide (5, 10, or 20 mg/kg; experiment 1), propranolol (10 or 20 mg/kg; experiment 2) or saline. Using the minimally effective doses from the light enhanced startle experiments, conditioned fear to a shock-paired cue was also evaluated. In a third experiment, rats were (a) removed from the test cages and briefly handled between phases I and II, (b) were not handled during this interval, or (c) were tested without the interposed delay. RESULTS: Propranolol (10 and 20 mg/kg) and chlordiazepoxide (10 mg/kg) disrupted light-enhanced startle at doses comparable to those required to disrupt fear-potentiated startle to a shock-paired cue. There was no effect of handling. CONCLUSION: These results further characterize the pharmacology of light-enhanced startle, provide additional support for the view that the effects of light on startle reflect an influence of anxiety, and offer additional information concerning the procedural variables that influence this behavior. PMID- 11862364 TI - The effects of noradrenergic re-uptake inhibition on memory encoding in man. AB - RATIONALE: Animal and human evidence implicate the central noradrenergic system in the process of memory modulation for emotional material. Blockade of the beta adrenergic system in humans has been shown to result in decreased recall and recognition memory performance, relative to placebo, for the emotional elements of a series of slides accompanied by a narrative. Stimulation of the noradrenergic system with yohimbine has also been shown to result in increased recall and recognition performance relative to placebo for the same stimulus materials. OBJECTIVES: The present study tested the hypothesis that stimulating the central noradrenergic system using the new noradrenaline re-uptake inhibitor reboxetine would result in a dose-dependent enhancement of memory for emotional material in man. METHODS: The central noradrenergic system was manipulated using reboxetine in a double-blind, randomised between-group, placebo-controlled design with 36 healthy adult subjects in each of three groups (placebo, 4 and 8 mg reboxetine). Free recall and recognition memory performance were assessed in a 'surprise' memory test following a 7-day interval. RESULTS: We found no memory enhancing effect of reboxetine. In contrast we observed a dose-dependent effect on memory opposite to the predicted direction. There were no significant differences between groups in self-rated stress and arousal scores or self-rated emotional reactions to the stimuli. All groups showed the expected increased memory performance for the middle 'emotive' phase of the story. CONCLUSION: Selective stimulation of the central noradrenergic system at encoding did not result in enhanced long-term memory for emotional material in man. PMID- 11862365 TI - A randomized controlled trial of high dose ascorbic acid for reduction of blood pressure, cortisol, and subjective responses to psychological stress. AB - RATIONALE: Physiological responses to stress are considered disruptive to health. High-dose ascorbic acid has reduced indices of stress in laboratory animals. METHODS: We conducted a randomized double-blind, placebo-controlled 14-day trial of sustained-release ascorbic acid (60 healthy young adults; 3 x1000 mg/day Cetebe) and placebo (60 healthy young adults) for reduction of blood pressure, cortisol, and subjective response to acute psychological stress (Trier Social Stress Test, TSST, consisting of public speaking and mental arithmetic). Six subjects from each group were excluded. RESULTS: Compared to the placebo group, the ascorbic acid group had less systolic blood pressure (an increase of 23 versus 31 mmHg), diastolic blood pressure, and subjective stress responses to the TSST; and also had faster salivary cortisol recovery (but not smaller overall cortisol response). Cortisol response to 1 microg ACTH, and reported side-effects during the trial did not differ between groups. Plasma ascorbic acid level at the end of the trial but not pre-trial was associated with reduced stress reactivity of systolic blood pressure, diastolic blood pressure, and subjective stress, and with greater salivary cortisol recovery. CONCLUSIONS: Treatment with high-dose sustained-release ascorbic acid palliates blood pressure, cortisol, and subjective response to acute psychological stress. These effects are not attributable to modification of adrenal responsiveness. PMID- 11862367 TI - Use of amphetamine by recreational users of ecstasy (MDMA) is associated with reduced striatal dopamine transporter densities: a [123I]beta-CIT SPECT study- preliminary report. AB - RATIONALE: Tablets sold as ecstasy often contain not only 3,4 methylenedioxymethamphetamine (MDMA) but other compounds well known to cause dopaminergic neurotoxicity, such as (meth)amphetamine. Furthermore, the use of ecstasy in the Netherlands is often combined with the use of amphetamine. However, little is known about the effects of ecstasy use or the combination of ecstasy and amphetamine use on dopamine (DA) neurones in the human brain. OBJECTIVES: This study was designed to investigate the effects of ecstasy as well as the combined use of ecstasy and amphetamine on the density of nigrostriatal DA neurones. METHODS: [123I]beta-CIT SPECT was used to quantify striatal DA transporters. Striatal [123I]beta-CIT binding ratios of control subjects ( n=15) were compared with binding ratios of ecstasy users ( n=29) and individuals with a history of combined ecstasy and amphetamine use ( n=9) after adjustment for age. RESULTS: Striatal [123I]beta-CIT binding ratios were significantly lower in combined ecstasy and amphetamine users compared to sole ecstasy users (6.75 versus 8.46, respectively: -20.2%, P=0.007). Binding ratios were significantly higher in ecstasy users when compared to controls (8.46 versus 7.47, respectively: +13.2%, P=0.045). CONCLUSIONS: These initial observations suggest that the sole use of ecstasy is not related to dopaminergic neurotoxicity in humans. In contrast, the reported use of amphetamine by regular users of ecstasy seems to be associated with a reduction in nigrostriatal DA neurones. PMID- 11862366 TI - Cues paired with a low dose of alcohol acquire conditioned incentive properties in social drinkers. AB - RATIONALE: Drug-related cues may acquire incentive properties through classical conditioning. OBJECTIVE: The present study investigated whether arbitrary stimuli paired with a low dose of alcohol would evoke differential skin conductance, salivary, craving and attentional orienting responses compared to arbitrary stimuli paired with vehicle in a sample of social drinkers. METHODS: A discriminative classical conditioning procedure was employed, in which subjects repeatedly consumed two drinks, which differed in terms of the flavour of the drink and the colour of the glass in which it was administered. One of the drinks (CS+) always contained 0.2 g/kg ethanol in flavoured tonic water and the other drink (CS-) always contained flavoured tonic water only. Alcohol craving, amount of salivation, and skin conductance level were measured in response to CS+ and CS during conditioning training. After conditioning training, the frequency with which subjects directed their attention to CS+ and CS- drinks and their choice of drinks was assessed. RESULTS: Comparable taste intensity ratings were given for alcohol and placebo drinks. Higher ratings of craving in the presence of CS+ and higher feelings of "lightheaded", "relaxed" and "contented" after consumption of the CS+ drink (compared with CS-) were observed over successive conditioning sessions. Skin conductance level was also higher in response to CS+ compared to CS-. After conditioning sessions were completed, subjects shifted their gaze more frequently towards CS+ than CS-, although they did not choose to consume more CS+ than CS- drinks when instructed to choose between them. CONCLUSIONS: These data suggest that conditioned responses to an arbitrary cue paired with alcohol will develop rapidly and provide further support for incentive salience theories of drug addiction. PMID- 11862368 TI - Long-term effects of neonatal MK-801 treatment on spatial learning and cortical plasticity in adult rats. AB - RATIONALE AND OBJECTIVES: The long-term effects of neonatal treatment with MK-801 on spatial learning and cortical plasticity were investigated in adult rats. METHODS: Rat pups were injected twice daily with MK-801 (0.1 mg/kg) on postnatal days 7-19, participated in water maze testing between postnatal days 90 and 102, and were then studied electrophysiologically. RESULTS: Treatment with MK-801 in such a low dose resulted in a very slight impairment of performance in the water maze task, but not in the visual cue response. Besides the slight learning impairment, the electrophysiological study revealed a reduction in the capacity for plasticity in the primary motor cortex of the treated animals, which was pronounced in the controls. CONCLUSION: The study demonstrates that even a slight impairment in learning and memory function may be accompanied by a cortical plasticity deficiency that is detectable electrophysiologically. PMID- 11862369 TI - Effects of amphetamine on the plus-maze discriminative avoidance task in mice. AB - RATIONALE: The contradictory amphetamine effects on memory could be due to different protocols of amphetamine administration or the well-known anxiogenic effect of the drug. OBJECTIVE: The effects of different protocols of administration of amphetamine were investigated on mice tested in the plus-maze discriminative avoidance task (DAT), which provides simultaneous information about memory and anxiety. METHODS: Acutely pre- or post-training, 0.3, 1.0, or 3.0 mg/kg amphetamine-treated, 10-day chronically 3.0 mg/kg amphetamine-treated, 0.3 mg/kg amphetamine plus 0.25 mg/kg scopolamine and 3.0 mg/kg amphetamine plus 3.0 mg/kg tacrine-treated mice were conditioned to choose between two enclosed arms (one of which was aversive) while avoiding two open arms. Learning/memory was evaluated by the percentage time in the aversive enclosed arm (PTAV), and anxiety by the percentage time in the open arms (PTO). RESULTS: Given acutely before conditioning, amphetamine significantly decreased PTO in training, suggesting an anxiogenic effect, and significantly increased PTAV in the test, suggesting an amnestic action. Given acutely after the conditioning, no action of this drug on memory was found. After repeated treatment, the anxiogenic effect disappeared, while the amnestic effect remained. While no effects of subeffective doses of amphetamine and scopolamine co-administration were detected, tacrine attenuated the amnestic effect of amphetamine. CONCLUSIONS: Amphetamine has different effects on DAT when given pre- or post-training. While acute pre training amnestic action is temporally correlated with an anxiogenic effect, there is tolerance to the anxiogenic but not to the amnestic effect after repeated administration. Because this acute amnestic effect of amphetamine is attenuated by tacrine, a possible relationship with cholinergic system cannot be discarded as a mechanism to amphetamine-induced amnesia in DAT. PMID- 11862370 TI - Naltrexone decreases craving and alcohol self-administration in alcohol-dependent subjects and activates the hypothalamo-pituitary-adrenocortical axis. AB - BACKGROUND: This laboratory study investigated the mechanisms by which the opioid antagonist, naltrexone, reduces the risk of relapse to heavy drinking in individuals with alcohol dependence. METHODS: Eighteen alcohol-dependent, non treatment-seeking volunteers were randomized to 50 mg naltrexone or placebo for 6 days and participated in an alcohol self-administration experiment on the sixth day. Following baseline assessments of craving and endocrine levels, subjects were first administered a priming drink designed to raise blood alcohol levels to 0.03 g/dl and then had the opportunity to drink up to eight additional drinks or to receive US $3 for each drink not consumed over a 2-h period. Each additional drink was designed to raise blood alcohol levels by 0.015 g/dl. RESULTS: At baseline, naltrexone treatment resulted in higher cortisol levels and lower levels of craving than placebo treatment. Although there were no significant differences in response to the priming dose, naltrexone-treated subjects drank fewer drinks, consumed them more slowly, and reported lower levels of alcohol craving during the alcohol self-administration portion of the experiment. Naltrexone also resulted in higher levels of adrenocorticotropic hormone and cortisol than placebo treatment, and levels of cortisol were negatively correlated with intensity of alcohol craving. The number of drinks chosen was positively correlated with level of alcohol craving. Ratings of nausea were low and did not differ between the naltrexone and placebo groups at any point in the study. CONCLUSIONS: These results confirm the hypothesis that naltrexone reduces desire to drink and the amount of alcohol consumed in alcohol-dependent subjects. It is hypothesized that naltrexone may reduce drinking via suppressing craving for alcohol and that this effect may be related in part to naltrexone's ability to activate the hypothalamo-pituitary-adrenocortical axis. PMID- 11862371 TI - Anxiolytic-like effects of AP7 injected into the dorsolateral or ventrolateral columns of the periaqueductal gray of rats. AB - RATIONALE: Glutamate antagonists microinjected into the dorsolateral PAG (DLPAG) show an anxiolytic-like profile in the elevated plus maze. Other columns of the PAG are also involved in defensive reactions. Few studies, however, have investigated the effects of pharmacological manipulation of the ventrolateral PAG (VLPAG) on procedures that predict anxiolytic activity. OBJECTIVES: To investigate the effects of the NMDA receptor (NMDAr) antagonist 2-amino-7 phosphonoheptanoic acid (AP7) microinjected into the DL or VLPAG in two procedures that predict anxiolytic activity using distinct aversive contingencies, the elevated plus maze and the Vogel punished licking test. METHODS: Male Wistar rats (7-14/group) with cannulas aimed at the DLPAG or VLPAG received AP7 (2 nmol/0.5 microl) or saline and 10 min later were submitted to the behavioural tests. In the punished licking experiment, water deprived (48 h) animals were allowed to drink for 3 min, receiving a 0.5 mA shock every 20 licks. The elevated plus maze test was performed as described elsewhere. Using this test, a dose response-curve for AP7 (0.2-20 nmol) injected in a smaller volume (0.25 microl) into the VLPAG was also performed. RESULTS: AP7 increased exploration of open arms of the EPM when microinjected into either the DLPAG or VLPAG ( P<0.05, ANOVA). The drug also increased the number of punished licks when administered into those columns (ANOVA, P<0.05). CONCLUSIONS: The results suggest that antagonism of endogenous excitatory amino acid neurotransmission in the DLPAG or VLPAG is able to reverse behavioral suppression induced by distinct aversive contingencies. PMID- 11862372 TI - Repeated alcohol: behavioral sensitization and alcohol-heightened aggression in mice. AB - RATIONALE: Repeated administration of psychomotor stimulants or opiates can induce behavioral sensitization, typically detected as progressive and long lasting increases in the motor-activating effects of these drugs. This phenomenon may be relevant to seizure susceptibility, drug self-administration, and sexual behavior. Repeated administration of alcohol can also induce behavioral sensitization and may have consequences on how alcohol affects aggressive behavior. OBJECTIVES: To (1) determine the enduring nature of locomotor sensitization to alcohol; (2) examine subsequent changes to morphine and amphetamine effects on locomotor behavior; and (3) test whether behavioral sensitization to alcohol or morphine is relevant to alcohol-heightened aggression. METHODS AND RESULTS: In the first experiment, male CFW mice were given ten injections of alcohol (2.4 g/kg/day), morphine (30.0 mg/kg/day), or saline. Video tracking confirmed locomotor sensitization--an approximate 200% increase in the motor-stimulating effects of these drugs. Challenges with 2.0 g/kg alcohol revealed that locomotor sensitization to alcohol persisted for at least 2 months. Alcohol-sensitized mice showed evidence of cross-tolerance to the sedative effects of morphine (5 mg/kg) but showed no evidence of cross sensitization to the stimulant effects of 30.0 mg/kg morphine or 1.0 mg/kg amphetamine. In the second experiment, under conditions resulting in species typical aggressive behavior against a male intruder, there were no differences in the aggressive behavior relative to saline control mice following alcohol or morphine sensitization. However, in the mice sensitized to alcohol, but not to morphine, there was a vertical shift in the dose-effect curve for moderate doses of alcohol (0.6-1.7 g/kg, p.o.). In addition, twice as many alcohol-sensitized mice consistently showed alcohol-heightened aggression when compared with the saline control mice (74% vs 37%, respectively). CONCLUSIONS: Repeated administration of alcohol can sensitize locomotor stimulation and may also render mice more vulnerable to increased aggression after alcohol. Moreover, the results suggest that at least some of the neuroadaptations caused by repeated administration of alcohol are relevant to alcohol-heightened aggression. PMID- 11862373 TI - The GABA(B) antagonist CGP56433A attenuates the effect of baclofen on cocaine but not heroin self-administration in the rat. AB - RATIONALE: Several reports have demonstrated that the gamma-aminobutyric acid (GABA)(B) agonist baclofen attenuates the reinforcing effects of cocaine in rats, and recent evidence indicates that it might have a similar effect on heroin self administration. OBJECTIVES: The specific GABA(B) receptor antagonist CGP56433A was used to further evaluate the involvement of GABA(B) receptors in the baclofen induced suppression of cocaine and heroin self-administration. METHODS: In the first series of experiments, dose-response curves were generated to examine the effect of CGP56433A (0.6, 1.0, or 1.8 mg/kg, i.p.) on cocaine (1.5 mg/kg per injection) and heroin (25 microg/kg per injection) self-administration reinforced under a fixed-ratio (FR1) or progressive ratio (PR) schedule. Separate sets of experiments then examined the effect of the co-administration of CGP56433A and baclofen on responding for cocaine or heroin under both schedules. RESULTS: Pretreatment with CGP56433A had no effect on cocaine or heroin self administration, while baclofen dose dependently reduced responding for both cocaine and heroin under both the FR1 and PR schedule. CGP56433A (1.8 mg/kg) blocked the effect of baclofen on cocaine but not on heroin self-administration. CONCLUSION: The specific GABA(B) antagonist CGP56433A attenuated the effect of baclofen on cocaine self-administration, suggesting that GABA(B) receptors are critical in mediating the anti-cocaine effect of baclofen. In combination with other studies, the data demonstrate that the susceptibility of baclofen and other GABA(B) agonists to receptor blockade depends on the behavioral response being studied. Whether this indicates different receptor mechanisms are involved (e.g., pre- versus post-synaptic effects or differential receptor reserve) remains to be determined. PMID- 11862374 TI - Nicotine potentiation of brain stimulation reward reversed by DH beta E and SCH 23390, but not by eticlopride, LY 314582 or MPEP in rats. AB - RATIONALE: Systemic nicotine administration increases dopamine and glutamate levels in reward-related brain areas. Nicotine-induced increases of dopamine in the nucleus accumbens are in part mediated by glutamatergic projections to the ventral tegmental area dopamine neurons. OBJECTIVES: To assess the effects of actions at acetylcholine, dopamine, presynaptic (mGluR(2/3)) and postsynaptic (mGluR(5)) metabotropic glutamate receptors (mGluRs) on the potentiation of brain stimulation reward induced by systemically administered nicotine (0.125-0.5 mg/kg; free base) in rats. METHODS: A discrete-trial current-threshold s stimulation reward procedure (electrodes placed in the posterior lateral hypothalamus) was used to assess the effects of DH beta E (0.5-5 mg/kg), an acetylcholine nicotinic receptor antagonist, SCH 23390 (1.25-5 microg/kg), a dopamine D(1) receptor antagonist, eticlopride (2.5-20 microg/kg), a dopamine D(2) receptor antagonist, LY 314582 (1-20 mg/kg), an mGluR(2/3) agonist, and MPEP (1-9 mg/kg), an mGluR(5) antagonist, on the reward potentiating effects of nicotine (0.25 mg/kg). RESULTS: DH beta E had no effect on reward thresholds when administered alone, but dose-dependently reversed the nicotine-induced potentiation of brain stimulation reward. SCH 23390 (5 microg/kg) elevated thresholds when administered alone, and reversed the nicotine-induced potentiation of brain stimulation reward even at a dose (2.5 microg/kg) that had no effect on reward thresholds. Eticlopride (10-20 microg/kg), LY 314582 (10-20 mg/kg) and MPEP (9 mg/kg) elevated thresholds when administered alone but had no effect on the nicotine-induced potentiation of brain stimulation reward. CONCLUSIONS: These results indicate that nicotinic and dopamine D(1) receptors are involved in the nicotine-induced potentiation of brain stimulation reward, while actions at dopamine D(2), mGlu(2/3) and mGlu(5) receptors did not modulate this effect of nicotine. PMID- 11862375 TI - Diagnosis at the first episode to differentiate antidepressant treatment responses in patients with mood and anxiety disorders. AB - RATIONALE: Co-morbidity of mood and anxiety disorders is often ignored in pharmacotreatment outcome studies and this complicates the interpretation of treatment response. The clinical trials are usually based on single categories from the Diagnostic and Statistical Manual of Mental Disorders (DSM). OBJECTIVES: The present study is a first attempt to differentiate the responses to antidepressants using a design that differs from that used in previous clinical trials. To avoid bias due to co-morbidity, we included patients with any DSM-III R diagnosis of mood or anxiety disorder for which antidepressant treatment was indicated. We also explored the role of the diagnosis at the first episode in the efficacy of the different antidepressants. METHODS: A total of 92 outpatients with a mood and/or anxiety disorder were randomly assigned to treatment with imipramine or fluvoxamine in a 6-week study. The diagnosis at the first episode- or primary diagnosis--was available for 78 patients, 40 with a primary depression and 38 with a primary anxiety disorder. RESULTS: Analyses using the MIXED procedure for repeated measures showed no general differences between treatment with imipramine and treatment with fluvoxamine. When the primary diagnoses were taken into consideration, differentiation occurred. Patients with primary depression showed better responses to imipramine than to fluvoxamine. The assumption that patients with primary anxiety disorder would respond better to fluvoxamine than imipramine was observed for only the Clinical Global Impression. CONCLUSIONS: The results suggest that the nature of the first illness episode may be more valuable than the DSM categories of mood or anxiety disorders, which may lend support to the concept of primary versus secondary depression for purposes of differentiating treatment responses. Given the exploratory nature of the study, however, replication of our finding is needed. PMID- 11862376 TI - Effects of repeated cortisol administration on brain potential correlates of episodic memory retrieval. AB - RATIONALE: Neuropsychological impairments in depressive illness may be secondary to hypercortisolaemia. Cortisol administration to healthy subjects impairs episodic memory, though how this is mediated is unknown. OBJECTIVES: To examine the effects of 1 week's administration of cortisol on the neural correlates of episodic memory in healthy subjects. METHODS: Fourteen healthy men were treated with oral cortisol (hydrocortisone 20 mg) or placebo twice daily for 1 week, in a double blind, crossover fashion. Event related potentials (ERPs) were recorded during a well-validated source memory task. Subjects listened to words spoken in a male or female voice. At test, old and new words were presented visually; subjects judged whether words were old or new, and if old, the gender of the voice at study. RESULTS: Response times were significantly speeded by cortisol. A significant reduction in recognition accuracy with cortisol was found for the second study occasion. ERP recordings with placebo showed greater positivity over left parietal and right frontal scalp areas for ERPs to items given correct source judgements versus correctly rejected new items. In comparison, cortisol increased ERP voltage between 500 and 1400 ms post-stimulus and this effect interacted with item type and electrode site, being diffusely distributed for correct rejections but of a lesser magnitude frontally for old items accorded a correct source judgement. CONCLUSIONS: Repeated cortisol administration leads to a qualitative change in the neural correlates of episodic memory retrieval in healthy subjects. This change may contribute to cognitive impairments seen in illnesses characterised by hypercortisolaemia. PMID- 11862377 TI - Effect of sodium deprivation on morphine-and lithium-induced conditioned salt avoidance and taste reactivity. AB - RATIONALE: When paired with morphine, rats suppress their intake of saccharin solution, but not a less palatable salt solution. The reward comparison hypothesis argues that when a taste is paired with morphine, intake of the solution is expected to decrease as the palatability of the taste increases. Therefore, morphine should more effectively suppress intake of salt solution in rats that are conditioned in a sodium-deprived state than in rats that are conditioned in a sodium-replete state. OBJECTIVES: The present experiments evaluated the effect of furosemide-induced sodium deprivation on morphine and lithium-induced salt (experiment 1) and saccharin (experiment 2) avoidance and salt taste reactivity (experiment 4). METHODS: Rats were injected with furosemide or saline 21 h prior to access to salt solution (experiments 1 and 3) or saccharin solution (experiment 2). Immediately following access to the solution, the rats were injected with saline, morphine or lithium chloride solution. In experiments 1 and 2, a two-bottle test measured the strength of the taste preference/avoidance. In experiments 3 and 4, the taste reactivity test evaluated the furosemide-induced unconditional palatability changes for salt solution (experiment 3) and the conditional palatability changes for salt previously paired with morphine or lithium (experiment 4). RESULTS: Sodium depletion induced by furosemide pretreatment conditionally enhanced subsequent preference for salt solution using both the taste avoidance test (experiment 1) and the taste reactivity test (experiment 4). Salt-lithium associations, but not salt-morphine associations, suppressed salt preference. However, the salt-morphine (40 mg/kg) association enhanced salt preference (in both experiments 1 and 4) when rats were conditioned in a sodium-deprived state. In experiment 2, morphine-saccharin associations resulted in conditioned saccharin avoidance regardless of pretreatment condition. CONCLUSIONS: When the palatability of salt was enhanced by sodium depletion, morphine produced a mild conditioned salt preference in both a two-bottle preference test and enhanced ingestion reactions in the taste reactivity test, but morphine produced conditioned saccharin avoidance. PMID- 11862378 TI - Stereoselective effects of methylphenidate on motor hyperactivity in juvenile rats induced by neonatal 6-hydroxydopamine lesioning. AB - RATIONALE: The psychostimulant dl-threo-methylphenidate is commonly used to treat attention deficit-hyperactivity disorder (ADHD). Consistent with its effects in ADHD patients, racemic methylphenidate antagonizes behavioral hyperactivity in several animal models of ADHD, including juvenile rats with neonatal 6 hydroxydopamine (6-OHDA) lesions of forebrain dopamine projections. The enantiomers of methylphenidate differ markedly in stimulant potency but have not been compared in the 6-OHDA lesion model. OBJECTIVE: Locomotor-inhibiting effects of methylphenidate enantiomers were compared in 6-OHDA-lesioned rats to test the hypothesis that d-methylphenidate is more potent than dl- and l-methylphenidate. METHODS: Selective dopamine lesions were made using 6-OHDA (100 microg, intracisternal, IC) on postnatal day (PD) 5 after desipramine (25 mg/kg, SC) pretreatment to protect noradrenergic neurons. Effects of d-, l- and dl-threo methylphenidate on locomotor activity of lesioned and sham control rats were quantified at PD 23-27. RESULTS: Lesioning yielded robust motor hyperactivity at PD 23-27. Both d- and dl-methylphenidate stimulated locomotor activity in intact rats, and inhibited activity in lesioned rats. l-Methylphenidate did not affect locomotor activity in either lesioned rats or controls. d-Methylphenidate (ED(50)=1.66 mg/kg) was 3.3 times more potent than dl-methylphenidate (ED(50)=5.45 mg/kg) in reducing locomotor hyperactivity in lesioned rats. In addition, pretreatment of lesioned rats with l-methylphenidate significantly reduced the motor inhibiting effects of d-methylphenidate. CONCLUSIONS: The more active enantiomer, as predicted, was d-methylphenidate, but the l-enantiomer interfered with its effects, suggesting that clinical potency of d methylphenidate may be more than twice that of the racemate. PMID- 11862379 TI - Effects of the putative antagonist NCS382 on the behavioral pharmacological actions of gammahydroxybutyrate in mice. AB - RATIONALE: Gammahydroxybutyrate (GHB) is an endogenous chemical found in the human brain that when administered systemically readily crosses the blood-brain barrier and produces behavioral effects. Some previously reported observations, including reports of specific antagonism by NCS382 (6,7,8,9-tetrahydro-5-[H] benzocycloheptene-5- ol-4-ylidene acetic acid), support the hypothesis that GHB is a neurotransmitter with its own receptor system. In addition to its uncertain physiological role, the recent interest in GHB has been engendered by its illicit use and abuse. OBJECTIVES: To further characterize the behavioral effects of GHB and to evaluate NCS382 for its potential antagonistic effects. METHODS: Following the administration of GHB alone and in combination with NCS382, mice were tested in a Functional Observational Battery (FOB) and for their effects on locomotor activity and on schedule-maintained behavior. Additionally, spontaneous and NCS382-precipitated withdrawal in rats chronically treated with GHB was examined. RESULTS: In the FOB, GHB generally produced depressant-like effects that were generally not reversed by NCS382. GHB also dose dependently reduced locomotor activity and rates of operant behavior, which were generally not reversed by co administrations with NCS382. Neither spontaneous nor NCS382-precipitated signs of physical dependence were observed following chronic GHB administration. CONCLUSION: GHB dose dependently produced depressant-like effects on learned and unlearned behavior. The putative GHB antagonist NCS382 failed to convincingly antagonize these effects. Physical dependence was not evident following spontaneous withdrawal or NCS382 challenge. Taken together, these results suggest that NCS382's ability to antagonize GHB's effects may be very limited. PMID- 11862380 TI - Clozapine can induce high dopamine D(2) receptor occupancy in vivo. AB - RATIONALE: Clozapine is a unique antipsychotic with very low propensity to cause motor side effects. In contrast to most other antipsychotics that block more than 70% of dopamine D(2) receptors at therapeutic doses, clozapine occupies less than 70%. Furthermore, even at maximum occupancy, 70% is not exceeded. Several mechanisms have been proposed as explanations for this low D(2) receptor occupancy, but clear evidence is limited. OBJECTIVES: In patient studies the data are limited by the dose-range that can be safely used; therefore, the aims of this study were to examine the maximum occupancy of dopamine D(2) receptors with up to 5.0 mg/kg of bolus injection of clozapine to non-human primates and to measure the time course of occupancy. METHODS: PET examination with [(11)C]raclopride was performed to measure the dopamine D(2) receptor occupancy in the striatum of two monkeys after the bolus injection of 0.2-5.0 mg/kg clozapine. [(11)C]raclopride was injected sequentially to follow the time course of occupancy up to 7 h after the clozapine injection. RESULTS: Dopamine D(2) receptor occupancy reached up to 83% after 5.0 mg/kg clozapine injection. Occupancy decreased with a half-life of 7.22 h after 5.0 mg/kg clozapine and 5.25 h after 1.0 and 2.0 mg/kg clozapine. CONCLUSIONS: Clozapine could occupy a high proportion of dopamine D(2) receptors. The time course of occupancy was relatively fast, with a half-life of several hours. PMID- 11862381 TI - Baboon immunoglobulin variable region heavy chains: identification of genes homologous to members of the human IGHV1-IGHV7 subgroups. AB - Baboons are increasingly used as animal models in studies that require the assessment of specific antibody responses. The complete analysis of these responses includes the characterization of antibody variable region genes and their usage. Therefore, we cloned and sequenced a total of 27 immunoglobulin variable region heavy chain (IGHV) genes using cDNA obtained from a single baboon (Papio cynocephalus anubis). Comparison of baboon and human IGHV sequences shows that the percent of identity varies between 86% and 96% at the nucleotide level and between 80% and 95% at the amino acid level. The various baboon IGHV genes can be grouped into one of seven subgroups corresponding to the human IGHV1-IGHV7 subgroups. The only baboon cDNA sequence belonging to the IGHV7 subgroup is homologous to the human IGHV7-81 gene, for which no transcripts have been described. Similarly to their human counterparts, all baboon IGHV genes cluster into three clans, with one clan including the IGHV1, IGHV5 and IGHV7 subgroups, one clan including the IGHV2, IGHV4 and IGHV6 subgroups, and one clan including the IGHV3 subgroup. These data indicate that baboons may represent valuable models to analyze the antibody repertoire generated in response to immunization strategies developed for human use. PMID- 11862382 TI - Three novel mutations of the CIITA gene in MHC class II-deficient patients with a severe immunodeficiency. AB - Four transacting genes, CIITA, RFXANK, RFX5, and RFXAP, control coordinate MHC II expression. In humans, defects in these genes result in the absence of MHC II expression and thus a combined immunodeficiency. CIITA is considered to be a master MHC II regulator and is responsible for the defect in complementation group A. Eight such affected families have been reported. We investigated the molecular basis of the defect in three patients in these families, all presenting a severe immunodeficiency. CIITA transcripts were detected in all three patients but in one at an abnormally low level. Three novel heterozygous mutations of CIITA were found in patients SP and RC. One SP CIITA allele contained a nonsense mutation, G2178A, leading to a premature stop codon and the other allele in SP was found not to be expressed. In patient RC, two in-frame deletions were detected: one of the nucleotides 3003-3084 corresponding to the exon coding from Leu(964)to Asp(991), in the paternal allele, and a CATdel3193-5 of the isoleucine codon at position 1027, in the maternal allele. Transfection of a CIITA-deficient cell line with the recombinant CATdel3193-5-CIITA cDNA revealed a loss of function for this mutant and retention of the protein in the cytoplasm. No mutations were detected in the 4.5-kb cDNA from patient OK but the level of CIITA transcript was found to be profoundly decreased. However, promoters III and IV were not affected. This last case represents the first described CIITA dysfunction due to putative mutation(s) in cis regulatory sequences of CIITA. PMID- 11862384 TI - Mutation detection and physical mapping of the CD11 gene cluster in association with inflammatory bowel disease. AB - A genetic component in the etiology of inflammatory bowel disease (IBD) has clearly been demonstrated by epidemiological and genetic linkage studies. Linkage to IBD on proximal Chromosome (Chr) 16p is well established and replicated. A stratification experiment showed that the recent identification of a disease gene on the q arm does not interfere with the approach on the p arm, and the linkage peak is still significant. Here we present a candidate gene study of the alpha integrins (CD11A-D) on Chr 16. The alpha integrins play a key role in inflammatory processes, including leukocyte adhesion and migration. Their genes are located on the p arm of Chr 16, and therefore represent excellent positional and functional candidates. Since the assignment of the CD11 genes in the genome was not clear, we performed physical, radiation hybrid, and fluorescent in situ hybridization mapping of the gene family. All CD11 genes map on Chr 16p11-12. CD11B-D are arranged in a gene cluster within 300 kb and CD11A is located about 2.5 Mb telomeric. Thirteen new single nucleotide polymorphisms (SNPs) and eight SNPs from databases were identified through full-length sequencing. Case-control statistics demonstrated an association lead in the CD11 gene cluster, which was not confirmed in further family based association/linkage analyses using single markers and haplotypes. It is unlikely that the CD11 genes play an important role in the pathogenesis of IBD. The marginally significant results could indicate a disease gene in the vicinity of the gene cluster. PMID- 11862383 TI - Disparate binding of chaperone proteins by HLA-A subtypes. AB - We examined chaperone association with subtypes of HLA-A68 differing at positions 116 and/or 70, and analyzed the surface expression of each A68 subtype. Our findings with A68 indicate that certain subtypes have inefficient association with the assembly complex and correspondingly high surface expression, dependent on the character of position 116. Specifically, poor association of A68 subtypes with the transporter associated with antigen processing correlated with a comparatively high level of W6/32(+) forms at the cell surface. This observation suggests that intracellular retention is a dominant function of the assembly complex and that natural differences in assembly complex interaction may dictate the level of surface expression of MHC class I molecules. We also found that position 116 was crucial for HLA-A68 subtype association with the assembly complex. Our data contrast with results we obtained previously with HLA-B7 in that an aspartic acid at position 116 abrogated chaperone association for HLA A68, whereas it increased association for HLA-B7. In total, HLA-A molecules exhibit natural allele-specific distinctions in chaperone association that correlate with differences in cell surface expression and with the identity of amino acid position 116. PMID- 11862386 TI - The promoter polymorphism of the IL-6 gene is associated with levels of antibodies to 60-kDa heat-shock proteins. AB - Elevated levels of antibodies to 60 kDa heat shock proteins are associated with severe coronary heart disease and carotid atherosclerosis. The presence of self hsp60-reacting antibodies can only be partially explained by microbial infections and induction by bacterial hsp65 proteins, since important differences (including the epitope specificity and complement activating ability) between hsp60 and hsp65 reacting antibodies have been shown. The aim of this study was to investigate the possible effects of genetic polymorphisms of different genes of proinflammatory cytokines on anti-hsp60 autoantibody levels. One hundred and seventy-six male blood donors were recruited and antibody levels to human hsp60 and Mycobacterium bovis hsp65 were determined by ELISA. Also in these donors, polymorphisms of the promoter of the IL-6 gene at position -174, the biallelic base exchange of the IL-1 beta gene at the -511 position and the IL-1 alpha gene at position -889 were investigated by PCR. A strong association between IL-6 -174 polymorphism and anti-hsp60 antibody levels was seen; the effect on anti-hsp65 antibody was less marked. Carriers of allele C at this position had significantly lower levels of anti-hsp60 and anti-hsp65 antibodies. A lack of associations between IL-1 beta and IL-1 alpha gene polymorphisms and antibody levels was detected. This is the first study in which associations between genetic polymorphisms and autoantibody levels have been described in healthy subjects. Further studies are needed to gain insight into the detailed mechanism of how the IL-6 gene polymorphism at position -174 influences anti-hsp60 autoantibody levels. PMID- 11862388 TI - Unique TCR beta-subunit variable gene haplotypes in Africans. AB - This study investigated polymorphisms of genes in two regions of the T-cell antigen receptor beta-subunit (TCRB) locus, including BV9S2P, and BV6S7 in a 5' linkage group, and BV8S3, BV24S1, BV25S1, BV18S1, BV2S1, BV15S1 and BV3S1 in a 3' linkage group. These loci have been genotyped in individuals from five regions in Africa, including The Gambia, Nigeria, Cameroon, Tanzania, and Zambia, and in individuals from northern Britain, northern India, and Papua New Guinea (PNG). In the 3' linkage group, 11 unique haplotypes were identified in the combined African populations; two equally frequent haplotypes represent the majority of African chromosomes. One haplotype was found in all four regions studied. This is the most frequent haplotype in the northern British, northern Indian and PNG populations. Although present, it is infrequent in the African populations. A North-South gradient in the frequency of a common African haplotype was observed. The distribution did not represent that of a known disease. Evidence suggests that malaria is not responsible for selection of these haplotypes. Overall, this study highlights large differences in the genetic constitution of the TCRB locus between Africans and other populations. PMID- 11862387 TI - IMGT gene identification and Colliers de Perles of human immunoglobulins with known 3D structures. AB - A new database, IMGT/3Dstructure-DB, was developed and implemented in the IMGT (international ImMunoGeneTics database) information system (http://imgt.cines.fr) to provide a unique expertised resource on immunoglobulin and T-cell receptor structural data. Corresponding protein sequences were annotated with IMGT tools, which allow the precise identification of the genes expressed in these proteins, and the description of framework and complementarity determining regions according to the IMGT standardized nomenclature and IMGT unique numbering. Two dimensional graphical representations of the V-DOMAINs, designated as Colliers de Perles, are automatically produced. A query Web interface allows interactive search of the IMGT/3D structure-DB data. In this article, IMGT gene identification and Colliers de Perles of human immunoglobulins with known 3D structures in the Protein Data Bank are presented. PMID- 11862385 TI - Identification and characterization of SF2000 and SF2001, two new members of the immune receptor SLAM/CD2 family. AB - The SLAM family of human genes currently consists of seven related members of the immunoglobulin superfamily, membrane-associated proteins, including CD150 (SLAM), CD244 (2B4), CD84, CD229 ( Ly-9), BLAME, CD48, and 19A. These genes are expressed to varying degrees in subsets of immune cells (T, B, natural killer, and myeloid cells) and may function as ligands or receptors. This set of genes, related to CD2 and CD58 on Chromosome (Chr) 1p98, are found clustered close together in the human genome on Chr 1q22. Four of these family members (CD150, CD244, CD84, CD229) contain conserved tyrosine motifs in their cytoplasmic tails that enable them to bind intracellular signaling molecules SAP and EAT-2. SAP is mutated in human X-linked lymphoproliferative disease (XLP), and studies in XLP patients have shown that improper signaling via molecules that bind SAP contributes to the disease. We have identified two new members of the SLAM family (SF), which we term SF2000 and SF2001, which are expressed in immune cells and map in the SLAM gene cluster. SF2001 does not contain SAP-binding motifs in its short cytoplasmic tail. SF2000, which is co-expressed with SAP in T cells, binds both SAP and EAT 2. The data suggest that signaling through SF2000, together with CD150, CD244, CD84, and CD229, is controlled by SAP and therefore contributes to the pathogenesis of XLP. PMID- 11862389 TI - Malaria is not responsible for the selection of TCR beta-subunit variable gene haplotypes in The Gambia. AB - Previous work has shown that a single haplotype of the T-cell antigen receptor beta-subunit (TCRB) locus is predominant in African populations. This is likely to be due to selection pressure for gene(s) that protect children against disease. This study has tested the hypothesis that malaria is the responsible selection pressure, due to its impact on child mortality. The haplotypes of BV8S3, BV2S1, BV15S1, and BV3S1 were determined in children suffering from severe malaria and unaffected adult controls. No significant difference between cases and controls was shown for any of the haplotypes studied. In addition, an insertion/deletion (INDEL) haplotype in the 5' region of the TCRB locus was investigated. Again no differences between the two groups were detected. Therefore, the evidence suggests that malaria is not responsible for haplotype selection in The Gambia. PMID- 11862390 TI - MICA polymorphism in South American Indians. AB - We have studied the MICA alleles of 196 unrelated subjects from three South American Indian tribes (Toba, Wichi and Terena). They are members of isolated tribes located in the Gran Chaco area in northeastern Argentina and in Mato Grosso do Sul in South Central Brazil. Of 55 previously known alleles, nine were observed in South American Indians, compared with 16 that were found in North American Caucasians, suggesting a more restricted allelic distribution of MICA in these tribes. In South American Indians, MICA*00201 was the most frequent allele, with a gene frequency of 33% in Toba, 47% in Wichi and 44% in Terena. MICA*00201, MICA*027 (external domain sequence like MICA*008/TM allele A5) and MICA*010 accounted for more than 90% of all the MICA genes in South American Indians. In North American Caucasians, MICA*00801 (*008/A5.1) accounted for 42% of the genes and was the most common allele. We observed a high degree of linkage disequilibrium between certain alleles of MICA and of HLA-B in the South American Indian populations. Phylogenetic trees constructed using gene frequencies of the transmembrane short tandem repeats in the populations reported here, and in other populations taken from published reports, suggest that South American Indians are more closely related to Asians than to Europeans. PMID- 11862391 TI - Beta-defensin 1 gene variability among non-human primates. AB - Defensins are a recently described family of peptides that play an important role in innate immunity. Recent studies have shown that defensins exhibit a broad spectrum of antimicrobial activities against bacteria and fungi. Three families have been identified so far in mammals, alpha-defensins, beta-defensins and theta defensins, presumably derived from a common ancestral defensin. A long-term study on the evolution of these multigene families among primates has been undertaken to investigate: (1) the degree of interspecific differentiation; (2) the genetic mechanisms responsible for the variability of these molecules; and (3) the possible role of different environmental factors in their evolution. Nucleotide sequences have been obtained from great and lesser apes, several African and Asian catarrhine monkeys and one New World monkey. A comparison of rates of synonymous and nonsynonymous (amino-acid changing) nucleotide substitution indicates that the primate beta-defensin 1 gene evolved under a pattern of random nucleotide substitution as predicted by the neutral theory of molecular evolution. These results are not consistent with the hypothesis that the primate beta-defensin 1 gene has diversified in response to changes in the microbial species to which a given host is exposed. Analyses of interpecific variability have yielded some insights about the pattern of molecular evolution of the gene among primates. Humans and great apes present high levels of sequence similarity, differing in only one amino acid residue in the mature peptide. Compared with these taxa, hylobatids and cercopithecids exhibit 3-4 amino acid substitutions, some of which increase the net charge of the active molecule. PMID- 11862392 TI - Lack of a consistent relationship between demethylation of the CD44 promoter and CD44 expression. AB - Methylation of the CD44 promoter at one or more CpG dinucleotides has been proposed as being important in the control of CD44 expression. In vitro methylation of all CpGs occurring between bases -1 to -1374 upstream of the position of translation initiation repressed the promoter activity of mouse CD44 4.5- to 12-fold in transient transfection experiments. Assaying the methylation of these 29 CpGs by genomic sequencing using differential base modification by sodium bisulfite indicated that a cluster of three CpGs immediately upstream of the position of translation initiation was heavily methylated in the mouse CD44 negative T-cell lymphoma AKR1. All 19 CpGs between bases -4 and -447, including the cluster heavily methylated in AKR1, were demethylated in the CD44-positive T cell lymphoma BW5147, while CpGs further upstream showed no change in methylation pattern. The cluster of heavily methylated CpGs remained methylated, however, when CD44 was activated by transient transfection of c-jun into an AKR1 subline expressing polyoma large-T or by treatment of this subline with sodium butyrate, and no significant demethylation of other CpGs was observed. It is concluded, therefore, that no consistent demethylation event in the promoter-containing region encompassing the 1374 base pairs upstream of the position of CD44 translation initiation is required for CD44 expression. PMID- 11862393 TI - Chromosomal mapping of host susceptibility loci to Angiostrongylus costaricensis nematode infection in mice. AB - Angiostrongylus costaricensis is a nematode found mainly as a rodent parasite. Laboratory mice were experimentally infected with this parasite. It is known that there is great variability in mortality among inbred mouse strains after infection with this nematode. The survival rate at 5 weeks after infection of A/J mice was 90.5%, whereas that of SM/J mice was only 33.3%, with severe anemia and decreased body weight about 3 weeks after infection. To identify host susceptibility genes for infection with this nematode, we undertook chromosomal mapping by a whole-genome scanning approach in (A/JxSM/J)F2 mice. We mapped a host susceptibility locus (here designated Acsns, for Angiostrongylus costaricensis nematode susceptibility locus) to the telomeric portion of Chromosome 19 (peak LOD=4.35). We also identified two loci on Chr 13 and Chr 17 that have epistatic effects on host survival. This is the first report on host susceptibility loci for helminth infection mapped by whole-genome scanning. PMID- 11862394 TI - Nucleotide sequence of the MHC class I genomic region of a teleost, the medaka (Oryzias latipes). AB - Teleost orthologs of the human major histocompatibility complex (MHC)-encoded genes show a dispersed distribution over several chromosomal loci. However, some of them, including the MHC class I alpha chain genes, are tightly linked to each other, forming the teleost MHC class I region. To elucidate the evolution of vertebrate MHC, we have determined the complete nucleotide sequences of two bacterial artificial chromosome (BAC) clones, encompassing the MHC class I region of the inbred Hd-rR strain of the medaka (Oryzias latipes). The 425862 bp nucleotide sequence predicted 20 possibly expressed genes and three pseudogenes. Except for one gene, CIZ, whose human ortholog is located at 12p13.31, all identified genes were orthologs or closely related relatives of the human MHC encoded genes. Two classical class I alpha chain genes and the six other genes directly involved in class I antigen presentation formed an uninterrupted cluster. Comparison of the MHC class I region genes among three teleost species, the medaka, zebrafish and pufferfish, indicated that the content, but not the order or transcriptional orientation, of the genes is highly conserved. These results suggest that a strong selective pressure has conserved the linkage of certain MHC genes during vertebrate evolution, despite recurrent genetic rearrangements. PMID- 11862395 TI - Expansion and contraction of major histocompatibility complex genes: a teleostean example. AB - The MHC is a multigene family that has arisen through recurrent expansion and contraction of genes, and a continuum of the evolutionary process is observed in the teleost fishes. The number of duplicated genes observed in different phylogenetic groups of teleost fish varies from one to 42, with only a few genes observed in the primitive euteleost species, and greater numbers of genes observed in the more advanced neoteleost species. In this study, an attempt is made to isolate all of the Mhc class I genes of an early neoteleost species, Atlantic cod (Gadus morhua L.), in the superorder Paracanthopterygii. Eighty three sequences were isolated from the cDNA of an individual G. morhua. The level of gene duplication observed within each of the lineages and sublineages was similar, and most contained an estimated two to four duplicated genes. Mhc class I gene duplication in G. morhua was independent of, and possibly more recent than, extensive duplication in the Acanthopterygian superorder. Only limited contraction of Mhc genes is observed in G. morhua. A low level of haplotype diversity is observed, with most individuals containing at least one copy of each of the lineages tested. Divergence of the conserved N- and C-terminal residues of the antigen recognition site is observed, indicative of the initial stage of degeneration from classical to non-classical genes. However, most or all of the lineages are still polymorphic, and degeneration is present both within and among lineages. Thus, the outcome (i.e., which genes will remain classical) is as yet undetermined. PMID- 11862397 TI - Alu polymorphism within the MICB gene and association with HLA-B alleles. AB - We describe the finding of an Alu repeat dimorphism within the first intron of the MICB gene. The frequencies of the two AluyMICB alleles, AluyMICB*0(absence of insertion) and AluyMICB*1(presence of insertion), and their associations with the highly polymorphic HLA-B locus were determined for 51 human cell lines and for 109 and 200 Caucasians and northeastern Thais, respectively. Analysis of the AluyMICB and HLA-B allelic relationships revealed that AluyMICB*1 occurred at relatively low gene frequency (0.118-0.157) [corrected] but was strongly associated with HLA-B17 (HLA-B57,HLA-B58) and HLA-B13. The AluyMICB locus provides a useful dimorphic marker for investigations on the level of linkage disequilibrium between MICB, MICA, and HLA-B loci. PMID- 11862396 TI - Molecular evolution of the NF-kappaB signaling system. AB - The mechanisms of innate immunity in vertebrates show certain overall resemblances to immune mechanisms of insects. Two hypotheses have been proposed to explain these resemblances. (1) According to the evolutionary continuity hypothesis, innate immune mechanisms evolved in the common ancestor of vertebrates and insects and have been conserved since that time. (2) In the independent-evolution hypothesis, the mechanisms of innate immunity in vertebrates evolved independently from invertebrate immune mechanisms. Phylogenetic analysis of five gene families (Pelle, Rel, IkappaB, Toll, and TRAF) whose members are involved in NF-kappaB signaling in vertebrates and insects were used to decide between these hypotheses. The phylogenies of the Rel and TRAF families strongly supported independent evolution of immune functions in vertebrates and invertebrates, and, except for a possible case in the Pelle family, orthologous molecules having immune functions in both vertebrates and invertebrates were not found. The results suggest that NF-kappaB represents an ancient, generalized signaling system that has been co-opted for immune system roles independently in vertebrate and insect lineages. PMID- 11862398 TI - A CD45 polymorphism associated with abnormal splicing is absent in African populations. AB - The CD45 antigen is essential for normal antigen receptor-mediated signalling in lymphocytes, and different patterns of splicing of CD45 are associated with distinct functions in lymphocytes. Abnormal CD45 splicing has been recognized in humans, caused by a C77G transversion in the gene encoding CD45 (PTPRC). Recently the C77G polymorphism has been associated with multiple sclerosis and increased susceptibility to HIV-1 infection. These studies suggest that the regulation of CD45 splicing may be critical for the proper function of the immune system. Because of these data we examined the frequency of the C77G allele in African and Asian populations from countries with high or low prevalence of HIV infection. Here we report that the variant CD45 C77G allele is absent in African populations. We further show that populations living in the Pamir mountains of Central Asia have a very high prevalence of the C77G variant. PMID- 11862399 TI - MHC class I genes of the tree shrew Tupaia belangeri. AB - Two MHC class I cDNA sequences from the tree shrew (Tupaia belangeri), Tube-W01 and Tube-W02, have been isolated which are probably derived from classical class I genes. Expression of the tupaia class I genes was investigated in several organs, in particular the brain, in which slightly different amounts of class I transcripts are detectable in different areas. Gene tree analysis performed with Tube-W01 and Tube-W02, and including class I sequences derived from other orders, indicated that the tupaia sequences cluster differently from Primates, Carnivora, Artiodactyla, Perissodactyla, and Rodentia, but might be related to Lagomorpha class I genes. PMID- 11862400 TI - MICA-A5.1 allele is associated with atypical forms of celiac disease in HLA-DQ2 negative patients. AB - We selected 38 consecutive celiac disease (CD) patients (from a group of 316 consecutive CD patients) and 91 healthy blood donors, all of whom were HLA-DQ2 (DQA1*0501/DQB1*0201) negative, and investigated the presence of the classically associated alleles HLA-DQ8 and HLA-DRB4. We also studied the distribution of MICA transmembrane alleles in the two clinical forms of the disease. For this reason, these 38 DQ2-negative patients were subdivided into two groups: 18 typical CD patients and 20 atypical CD patients. No differences were found in the distribution of the DRB4 allele between DQ2-negative patients and controls. The HLA-DQ8 heterodimer (DQA1*03xx/DQB1*0302) was increased in CD patients (29%) compared with controls (10%), but no statistical differences were found. No differences were observed in the frequency of these alleles between either group of CD DQ2-negative patients. MICA-A5.1 was increased in atypical CD patients when compared with the typical forms of disease ( P(c)=0.03) and with healthy controls (P(c)=0.002). No other MICA allele was found to be significantly increased in the groups under study. The presence of MICA-A5.1 in atypical CD DQ2-negative patients may indicate a possible role of this allele in the development of CD. PMID- 11862402 TI - Is tapasin a modified Mhc class I molecule? PMID- 11862404 TI - Genetic studies of B-lymphocyte deficiency and mastocytosis in strain A/WySnJ mice. AB - The A/WySnJ mouse, but not the related A/J strain, has peripheral B-lymphocyte deficiency and mastocytosis. Minimally, two quantitative trait loci (QTLs) control the B-cell deficiency in (A/WySnJ x CAST/Ei)F2 intercross mice; one of them, Bcmd-1, mapped to Chromosome (Chr) 15. Several QTLs controlled the mastocytosis in this intercross, and it was not possible to determine whether any of them co-segregated with Bcmd-1. We have now mapped a second QTL controlling the B-cell deficiency, Bcmd-2, to Chr 4. Furthermore, we narrowed the map position of Bcmd-1 to <2.0 cM. Both QTLs have been confirmed through the construction of AW. Bcmd-1(c), AW. Bcmd-2(c), and AW. Bcmd-1(c)Bcmd-2(c) recombinant congenic strains. The Bcmd-1 locus is the major regulator of B-cell homeostasis, while Bcmd-2 is the minor regulator, and their effects are additive, as shown by splenic B-cells analysis in these congenic strains. In addition, Bcmd 2 or a linked locus controls mastocytosis, while Bcmd-1 does not, as indicated by splenic mast cell analysis in the congenic strains. Thus, the major genetic controls on B-cell homeostasis and mast cell homeostasis in A/WySnJ mice are asserted by distinct genes. PMID- 11862403 TI - MICA genetic polymorphism and linkage disequilibrium with HLA-B in 29 African American families. AB - The human major histocompatibility complex (MHC) class I chain-related gene A ( MICA) is located 46 kb upstream of HLA-B and encodes a stress-inducible protein which displays a restricted pattern of tissue expression. MICA molecules interact with NKG2D, augmenting the activation of natural killer cells, CD8(+) alpha beta T cells, and gamma delta T cells. MICA allelic variation is thought to be associated with disease susceptibility and immune response to transplants. We investigated MICA allelic variations and linkage disequilibrium with HLA-A, B, and DRB1 loci on 110 parental haplotypes from 29 African-American families. PCR/sequence-specific oligonucleotide probing (SSOP) was used to define MICA polymorphisms in exons 2, 3, and 4. Ambiguous allelic combinations were resolved by sequencing exons 2, 3, and 4. Exon 5 polymorphisms were analyzed by size sequencing. For HLA-A, B and DRB1 typing, low-resolution PCR/SSOP and allelic PCR/sequence-specific priming techniques were used. Twelve MICA alleles were observed, the most frequent of which were MICA*008, MICA*004, and MICA*002, with gene frequencies of 28.2, 26.4, and 25.5%, respectively. Thirty-eight HLA-B- MICA haplotypic combinations were uncovered, 22 of which have not been reported in the HLA homozygous typing cell lines from the 10th International Histocompatibility Workshop. Significant positive linkage disequilibria were found in 8 HLA-B- MICA haplotypes. Furthermore, haplotypes bearing HLA-B*1503, *1801, *4901, *5201, *5301, and *5703 were found to segregate with at least two different MICA alleles. Our results provide new data about MICA genetic polymorphisms in African Americans, which will form the basis for future studies of MICA alleles in allogeneic stem cell transplantation outcome. PMID- 11862405 TI - Chromosomal mapping of the host resistance locus to rodent malaria (Plasmodium yoelii) infection in mice. AB - The disease outcome in malaria caused by the protozoan parasite Plasmodium is influenced by host genetic factors. To identify host genes conferring resistance to infection with the malaria parasite, we undertook chromosomal mapping using a whole-genome scanning approach in cross-bred mice. NC/Jic mice all died with high parasitemia within 8 days of infection with 1 x 10(5) parasitized erythrocytes. In contrast, 129/SvJ mice all completely excluded malaria parasites from the circulation and remained alive 21 days after infection. We performed linkage analysis in backcross [(NC/Jic x 129/SvJ)xNC/Jic] mice. The Pymr ( Plasmodium yoelii malaria resistance) locus was mapped to the telomeric portion of mouse Chromosome (Chr) 9. This locus controls host survival and parasitemia after infection. The Char1 locus ( P. chabaudi resistance locus 1), controlling host survival and peak parasitemia in P. chabaudi infection, was previously mapped to the same region. This host resistance locus mapping to Chr 9 may represent a ubiquitous locus controlling susceptibility to rodent malaria. Elucidation of the function of this gene will provide valuable insights into the mechanism of host defense against malaria parasite infection. PMID- 11862407 TI - Identification, mapping, and genomic structural analysis of an immunoreceptor tyrosine-based inhibition motif-bearing C-type lectin from homozygous clones of rainbow trout (Oncorhynchus mykiss). AB - Utilizing a spleen-derived cDNA library and rapid amplification of cDNA 5' ends, we cloned a novel type II C-type lectin from two homozygous clones of rainbow trout. The cDNA is 2535 bp in length, and contains a 1017-bp open reading frame. From this sequence, a protein containing 339 amino acids (aa) was deduced. Using PSI-BLAST to search the GenBank database, the deduced protein is a C-type lectin, belonging to the type II membrane receptors. The protein contains four domains: an 87-aa N-terminal cytoplasmic domain, a 21-aa transmembrane domain, an 82-aa neck domain, and a 149-aa C-terminal C-type lectin domain. Two immunoreceptor tyrosine-based inhibition motifs (ITIMs) were located in the N-terminal cytoplasmic domain. RT-PCR results indicated that this gene is transcribed mainly in peripheral blood lymphocytes, spleen, kidney, and gill, and its expression in liver and intestine is weak. Monoclonal antibody 1.14 was used to isolate B cells from peripheral blood lymphocytes. Analysis revealed that this gene is highly expressed in B cells. Genomic DNA was amplified with long-template PCR and sequenced. The gDNA is 12.0 kb in length and contains nine exons and eight introns. The first intron of the genes from the OSU and AR clones differed in length. Based on this difference, the genotype of 69 doubled-haploid offspring of OSU and AR were screened. Subsequently, this gene was mapped on the rainbow trout linkage map to group XXI. Results of a Southern blot indicated that the gene ( TCL-2) exists as a single copy in the rainbow trout genome. The genomic structure, the deduced protein structure, the tissue expression pattern, as well as the phylogenetic analysis of the carbohydrate recognition domain based on the deduced amino acid sequence indicate that TCL-2 resembles CD72; however, the carbohydrate recognition domain sequences of TCL-2 and CD72 are highly diverged. PMID- 11862406 TI - Identification of the Gasa3 and Gasa4 autoimmune gastritis susceptibility genes using congenic mice and partitioned, segregative and interaction analyses. AB - BALB/c mice thymectomized on their third day of life develop a high incidence of experimental autoimmune gastritis (EAG) which closely resembles human chronic atrophic (type A, autoimmune) gastritis. Linkage analysis of (BALB/cCrSlcxC57BL/6)F2 mice previously demonstrated that the Gasa1 and Gasa2 genes on distal Chromosome (Chr) 4 have major effects on the development of EAG in this murine model, while other loci displayed a trend towards linkage. Here, we implemented partitioned chi(2)-analysis in order to develop a better understanding of the genotypes contributing to susceptibility and resistance at each linkage region. This approach revealed that linkage of Gasa1 and Gasa2 to EAG was due to codominant and recessive BALB/cCrSlc alleles, respectively. To identify additional EAG susceptibility genes, separate linkage studies were performed on Gasa1 heterozygotes and Gasa2 C57BL/6 homozygotes plus heterozygotes so as to minimize the effects of these disease genes. The enhanced sensitivity of these analyses confirmed the existence of a third EAG susceptibility gene (designated Gasa3) on Chr 6. Epistatic interactions between the Gasa2 EAG susceptibility gene and the H2 were also identified, and the presence of an H2 linked susceptibility gene (Gasa4) confirmed by analysis of H2 congenic mice. PMID- 11862408 TI - Cloning and characterization of cDNA clones encoding membrane-bound and potentially secreted major histocompatibility class I receptors from walleye (Stizostedion vitreum). AB - Major histocompatibility (MH) gene polymorphism has been used to type populations of humans, mice, and fish. Walleye ( Stizostedion vitreum) comprise an economically important fishery in Lake Erie, but whether those in the western basin form a single population or separate shoal- and river-breeding populations is not known. To develop MH gene markers for use in defining their population structure, we constructed a head kidney cDNA library from which five full-length class I heavy-chain clones were isolated and sequenced. Although they came in roughly three sizes, 1300, 1400, and over 2000 bp, the clones all exhibited a high degree of sequence similarity to each other and to known teleost MH class I cDNAs in the area encoding the extracellular domains, but showed dramatic differences in their transmembrane and cytoplasmic domains. One clone had an AG repeat that eliminated the hydrophobicity of the transmembrane domain, indicating that it may encode a secreted class I receptor. The other four clones encode three distinctly different cytoplasmic domains. The two clones that encode the same cytoplasmic domain resemble those of the known teleost MH class I sequences the most. Southern blotting indicated that there were four copies of the gene present in the walleye genome. Northern blotting showed that class I MH genes are expressed in most tissues and mRNAs of all three size classes can be detected. A preliminary survey of the polymorphism of these genes indicates that they will provide useful markers for differentiating fish stocks. PMID- 11862409 TI - Structure of the beta-chain (B29) gene of the chicken B-cell receptor and conserved collinearity with genes for potential skeletal muscle sodium channel and growth hormone. AB - Abstract. Reported here is the cloning of the gene for the beta-chain (Ig beta) of the chicken B-cell receptor for antigen. A cDNA library made from B lymphocyte derived DT40 cells was first screened with a beta-chain cDNA clone from rat. A full-length clone (1556 bp) was obtained encoding a 226-amino-acid beta-chain having 40% sequence identity with the highly conserved beta-chains present in mammals and producing mRNA of 1.7 kb. Clones containing the full B29 gene encoding chicken Ig beta were isolated and fully sequenced. As in mammals, the chicken B29 gene consists of six exons. The chicken B29 gene is somewhat larger (4336 bp) than that of rat (3.1 kb) and human (3.6 kb). Genome collinearity was found in the region of the chicken B29 gene. Genes for potential skeletal muscle sodium channel (2.2 kb upstream) and for growth hormone (2.3 kb downstream) were found in the same order and transcriptional orientation as in mammals, though the genes are more closely positioned in the chicken. PMID- 11862410 TI - Analysis of genetic regulation of chicken spontaneous autoimmune thyroiditis, an animal model of human Hashimoto's thyroiditis. AB - We analyzed the genetic regulation of spontaneous autoimmune thyroiditis (SAT) by means of crosses between Obese strain (OS) chickens with the disease, and healthy inbred chicken of line CB. Mononuclear cell infiltration of the thyroid was used as a criterion for the disease. We confirmed the existence of one recessive gene, regulating the susceptibility of the thyroid gland to autoimmune attack. From the frequency of progeny with the thyroid infiltration in backcross and F2 generations, we presume the existence of one or two additional dominant genes coding for abnormal reactivity of the immune system. The total number of genes regulating SAT is therefore a maximum of three. We attempted to identify disease specific transcripts responsible for the initiation of the disease using suppression subtractive hybridization of RNA prepared from OS and CB thyroid lobes, obtained from 3-day-old chicks. From forward and reverse subtractions, we recovered a fragment mixture in the range of 300 bp to 1.5 kb. In total, 768 clones were screened and 9 were sequenced. Four of them represent unknown sequences. Two, specific for OS thyroid, correspond to envelope genes of avian endogenous viruses (ev)-1, -3 and -6. The expressed product of an env gene could be iodinated in the thyroid gland and become involved in thyroglobulin metabolism. This may be another possible mechanism driving SAT, as iodine modulates SAT in the chicken. Further experiments will be required to prove this hypothesis. Two thyroid-specific clones had significant alignment to human thyroglobulin, and one clone to human coatomer protein. We analyzed, by RFLP and RNA dot blots, cosegregation between clones for the env gene of endogenous viruses as the most promising candidate for involvement in driving SAT; we did not find differences at the DNA and RNA levels. We can possibly therefore rule out a simple involvement of env genes with the initiation of disease. PMID- 11862411 TI - Localization to chicken chromosome 5 of a novel locus determining salmonellosis resistance. AB - Clear genetic differences in the susceptibility of chickens to visceral infection by Salmonella have been observed and it has been possible to identify resistant and susceptible lines of inbred chickens. We report here the results of experiments to map directly the gene(s) controlling this trait in chickens by examining crosses between highly susceptible and highly resistant lines. In the mapping panel, a region on chicken Chromosome (Chr) 5 was found to have a large effect on resistance, and this effect was observed in three separate resource populations. Mapping of additional marker loci in the region of the resistance gene further localized it to a region of approximately 2 cM, close to the genes for creatine kinase (CKB) and dynein (DNCH1). This region shows conserved synteny with telomeric regions of human Chr 14 and mouse Chr 12. On the basis of this conserved synteny, this resistance gene seems unlikely to correspond to the previously identified salmonellosis resistance genes Lps (located on mouse Chr 4) or Nos(2) (located on mouse Chr 11). There was no association between Nramp1 and resistance in these crosses, although this gene was shown to contribute to resistance in other crosses. The homologous human and mouse regions at present contain no likely candidate genes for this trait. Thus this appears to be a novel resistance gene, which we designate SAL1. PMID- 11862412 TI - Evolution of Mhc class II B genes in Darwin's finches and their closest relatives: birth of a new gene. AB - The 15 extant species of Darwin's finches on the Galapagos and Cocos Islands are the products of an unfinished adaptive radiation from a founder flock of birds related to the South American species Tiaris obscura. Molecular characterization of their major histocompatibility complex ( Mhc) class II B genes has revealed the existence of several related groups of sequences (presumably encoded in distinct loci) from which one (group 5) stands out because of its low divergence over extended time periods. Analysis of group 5 exon 2 and intron 2 sequences has revealed that the encoding locus apparently arose 2-3 million years ago in the Tiaris group of South and Central American Thraupini. The locus shows no evidence of inactivation, but displays a very low degree of polymorphism, both in terms of number of alleles and genetic distances between alleles. Some of the polymorphism, however, appears to be trans-specific. All the observed intergenic differences can be explained by point mutations and most of the exon 2 changes represent non-synonymous substitutions, although the rate of non-synonymous and synonymous substitutions appears to be the same. The origin of the new locus is explained by the birth-and-death model of Mhc evolution with two important extensions. First, the ancestor of the group 5 genes may have arisen without new gene duplication and second, the birth of the new group may have been brought about by a switch from balancing to directional selection. The ancestor of the group 5 genes may have been a classical class II B allele (one of many) which directional selection fixed in the ancestral population and drove into the category of nonclassical genes. PMID- 11862413 TI - Molecular and functional characterization of genes encoding horse MHC class I antigens. AB - Sequence and functional analyses were undertaken on two cDNAs and a genomic clone encoding horse major histocompatibility complex (MHC) class I molecules. All of the clones were isolated from a single horse that is homozygous for all known horse MHC class I and class II antigens. The two cDNAs (clones 8-9 and 1-29) were isolated from a lymphocyte library and encode polymorphic MHC antigens from two loci. The genomic cosmid clone, isolated from a sperm library, contains the 8-9 gene. All three genes were expressed in mouse L-cells and were recognized by alloantisera and, for the cDNAs, by alloreactive cytotoxic T lymphocytes. A total of 3815 bp of the genomic clone were sequenced, extending from 429 bp upstream (5') of the leader peptide through the 3' untranslated region. Promoter region motifs and an intron-exon structure characteristic of MHC class I genes of other species were found. A subclone containing 407 bp of the promoter region was inserted into a chloramphenicol acetyl transferase reporter plasmid, tested in transient transfection assays, and found to have promoter activity in heterologous cells. This genomic clone will enable detailed studies of MHC class I gene regulation in horse trophoblasts, and in horse retroviral infections. PMID- 11862414 TI - Polymorphism and chromosomal mapping of the mouse gene for B-cell activating factor belonging to the tumor necrosis factor family (Baff) and association with the autoimmune phenotype. PMID- 11862416 TI - Generation of anti-idiotypic reagents in the EGFRvIII tumor-associated antigen system. AB - The use of anti-idiotype (anti-id) vaccines for immunotherapy of human cancers is attractive, as immunization with true anti-id reagents (Ab2 beta) has been shown to induce both cellular and humoral immunity, frequently when the original antigen does not, or when a state of anergy to the self-expressed tumor associated antigen exists. The aim of this study was to investigate the potential of an anti-id vaccine approach to the glioma-associated antigen epidermal growth factor receptor variant III (EGFRvIII) for human clinical trials. By using conventional methodology, seven rat mAbs specific for the binding site of the murine anti-EGFRvIII-specific mAb Y10, as defined by the ability to inhibit the binding of mAb Y10 to EGFRvIII expressed on cells or as purified protein, were generated, and a subset (3/7) was found to be true Ab2 beta, as defined by the ability to induce the formation of antibody directed against EGFRvIII in two species (mouse and rabbit) when used as immunogen. The ability of these three Ab2 beta to elicit a protective anti-tumor response when used as a vaccine in the syngeneic, subcutaneous C57Bl/6-B16mseEGFRvIII tumor model was investigated. Following vaccination with one Ab2 beta mAb (2C7), 6/20 mice failed to develop tumor upon challenge, and 3/20 mice with outgrowing tumors exhibited dramatic regression of incipient tumors. Vaccination with a second mAb (5G8) resulted in one tumor-free survivor and one tumor regressor; vaccination with the third Ab2 beta mAb (7D3) did not confer protection, but did significantly increase the latency period until tumor outgrowth in all vaccinated recipients. The ability of Ab2 beta mAb 2C7 to induce an anti-EGFRvIII response in non-human primates was investigated by using the saponin adjuvant approved for human clinical trial, QS 21. Three of three macaques produced anti-EGFRvIII titers, as detected on EGFRvIII-expressing cells by both ELISA and fluorescence-activated cytometric analysis, following six immunizations with Ab2 beta mAb 2C7 and QS-21. The results obtained confirm that an anti-id response in the EGFRvIII antigen system can be induced in rodents, rabbits, and non-human primates, and it may prove a useful adjunct to immunotherapeutic approaches to EGFRvIII-positive gliomas, breast carcinomas, and non-small-cell lung tumors. PMID- 11862417 TI - Induction of intratumoral tumor necrosis factor by a synthetic lipid A analog, ONO-4007, with less tolerance in repeated administration and its implication in potent antitumor effects with low toxicity. AB - To evaluate the anti-tumor characteristics of ONO-4007, a synthetic analog of lipid A, the authors examined its acute toxicity and anti-tumor activity in a mouse MM46 mammary tumor system in comparison with LA-15-PP, an E. coli-type synthetic lipid A and LPS. Systemic and local (tumor site) induction of tumor necrosis factor (TNF) by a single i.v. shot of ONO-4007 and LA-15-PP correlated with manifestation of their toxicity, showing that ONO-4007 is 100-fold less effective than LA-15-PP. However, a protocol of repeated administration (3 shots twice a week) exhibited about 10 times more therapeutic potency of ONO-4007 for cancer therapy than expected in the above experiments. In a dose inducing submaximal systemic and intratumoral TNF production, repeated injections (twice a week) of ONO-4007 (10 mg/kg), LA-15-PP (0.1 mg/kg) and LPS (0.1 mg/kg) commonly generated a tolerant state in the systemic response (serum and liver) to subsequent stimulation. The intratumoral response was retained with this repeated administration of ONO-4007, but was not with LA-15-PP or LPS. TIM (tumor infiltrating macrophages) isolated from mice pre-injected with ONO-4007 and LA-15 PP were found to lose their response to both substances, but the response was rapidly recovered until 72 h after injection and virtually no difference was observed in their response to either drug. The in vitro treatment of naive TIM with ONO-4007 or LA-15-PP for 2 h depressed the response to both substances and the depression continued for 72 h even in culture with fresh medium. The relatively high efficacy of ONO-4007 in cancer therapy likely depends on the retraction of the tolerant state, especially at the tumor site where the response to ONO-4007 is recovered much more efficiently than that to lipid A. While constant recruitment of macrophages to tumor tissue might be involved in the difference of tolerance recovery between this region and others, selective response to ONO-4007 may not be explained simply by the sensitivity of recruited TIM. Pharmacokinetical experiments revealed that repeated injections of LA-15-PP enhanced its clearance from blood circulation, while the clearance of ONO-4007 was stable after repeated injections. Thus, pharmacokinetical properties of ONO 4007 may also possibly be implicated in this event. PMID- 11862419 TI - Tumor-associated antigen human chorionic gonadotropin beta contains numerous antigenic determinants recognized by in vitro-induced CD8+ and CD4+ T lymphocytes. AB - The beta subunit of human chorionic gonadotropin (hCG beta) is markedly overexpressed by neoplastic cells of differing histological origin including those present in colon, breast, prostate and bladder tumors. We have previously shown that some patients with hCG beta-producing urothelial tumors have circulating T cells that proliferate in response to hCG beta. To make a comprehensive study of hCG beta as a potential target for cancer immunotherapy, we investigated whether hCG beta peptides could induce CD4+ or CD8+ T-cell responses in vitro. By stimulating peripheral blood mononuclear cells (PBMCs) from three donors with mixtures of overlapping 16-mer synthetic peptides analogous to portions of either the hCG beta 20-71 or the hCG beta 102-129 region, we established six CD4+ T-cell lines that proliferated specifically in response to five distinct determinants located within these two hCG beta regions. Three antigenic determinants (hCG beta 52-67, 106-121 and 114-125) were presented by HLA-DR molecules, while the two other antigenic determinants (hCG beta 48-63 and 56-67) were presented by HLA-DQ molecules. Interestingly, one T-cell line specific for peptide hCG beta 106-121 recognized hCG beta peptides comprising, at position 117, either an alanine or an aspartic acid residue, with the latter residue being present within the protein expressed by some tumor cells. In addition, three other hCG beta-derived peptides that exhibited HLA-A*0201 binding ability were able to stimulate CD8+ cytotoxic T cells from two HLA-A*0201 donors. These three immunogenic peptides corresponded to regions hCG beta 40-48, hCG beta 44-52 and hCG beta 75-84. Our results indicate that the tumor-associated antigen hCG beta possesses numerous antigenic determinants liable to stimulate CD4+ and CD8+ T lymphocytes, and might thus be an effective target antigen for the immunotherapy of hCG beta-producing tumors. PMID- 11862420 TI - SV40 Tag-specific cytotoxic T lymphocytes generated from the peripheral blood of malignant pleural mesothelioma patients. AB - Malignant pleural mesothelioma (MPM) is an aggressive cancer, with survival of less than one year following diagnosis and treatment with current protocols. Recent studies have demonstrated the presence of the simian virus 40 (SV40)-like, large tumor antigen (Tag) in nearly 60% of MPMs. SV40 Tag is a viral-encoded tumor-specific antigen, and thus a potential target for the induction of anti tumor immunity and the development of therapeutic vaccines. We describe here evidence for the existence of SV40 Tag-specific immune responses in patients with MPM whose tumors express Tag. Humoral immunity was demonstrated by the detection of IgG titers against Tag in serum samples from 1/3 of patients examined. CTLs were generated from the peripheral blood of an HLA-A2(+) MPM patient with a synthetic peptide representing an HLA-A2 binding epitope in SV40 Tag. The CTLs demonstrated epitope fine specificity, in that other peptides from SV40 Tag and a peptide from influenza virus were not recognized in the context of HLA-A2. Moreover, the CTLs were capable of recognizing mesothelioma tumor cells that expressed SV40 Tag, in an MHC class I restricted manner. PMID- 11862418 TI - Sodium butyrate enhances complement-mediated cell injury via down-regulation of decay-accelerating factor expression in colonic cancer cells. AB - Decay-accelerating factor (DAF) expressed on the surface of colonic cancer cells presents a barrier to complement-mediated clearance by contributing to the ineffectiveness of the humoral immune response. In this study, to investigate the mechanisms responsible for the anti-tumor effects of butyrate, we evaluated how butyrate modulates DAF expression in colonic cancer cells. Three colonic cancer cell lines (HT-29, Caco-2, and T84 cells) were studied. DAF protein expression was assessed by western blot, and DAF mRNA expression was evaluated by northern blot. Complement C3 deposition on the surface of colonic cancer cells was determined by enzyme-linked immunosorbent assay (ELISA). The promoter activity of the DAF gene was assessed by a reporter gene-luciferase assay. Butyrate reduced the basal and interleukin-4 (IL-4)- and tumor necrosis factor-alpha (TNF-alpha) induced expression of DAF protein and mRNA in HT-29 cells. It increased the susceptibility to complement attack and enhanced C3 deposition on HT-29 cells. The inhibitory effect of butyrate on DAF mRNA expression was also observed in T84 and Caco-2 cells. Butyrate decreased basal DAF expression at both transcriptional and post-transcriptional levels. The inhibitory effect of butyrate on IL-4 induced DAF expression was closely associated with a blockade of IL-4-induced DAF mRNA stability. TNF-alpha-induced transcriptional activation and the increased stability of the DAF gene were also blocked by butyrate. Similar but weak effects were induced by trichostatin A, a potent histone deacetylase inhibitor, suggesting that histone acetylation might participate in butyrate activity. These observations indicate that both a down-regulation of DAF expression and the induction of susceptibility to complement attack contribute to the anti-tumor effects of butyrate in colonic cancer. PMID- 11862421 TI - Apoptotic pathways of cell death induced by an interleukin-13 receptor-targeted recombinant cytotoxin in head and neck cancer cells. AB - Interleukin 13 receptor (IL-13R)-targeted cytotoxin, IL13-PE38QQR, composed of IL 13 and a mutated form of Pseudomonas exotoxin (PE), is found to be highly and specifically cytotoxic to human solid cancer cell lines. However, the mechanism of tumor cell death mediated by IL-13 toxin is still not known. To elucidate the mechanism, we utilized four head and neck cancer cell lines (SCC-25, HN12, KCCT873, and YCUM911), which express high levels of IL-13R, and IL-13 toxin is highly cytotoxic to these cells. We observed chromatin condensation and DNA fragmentation, indicating apoptotic cell death, after treatment with IL-13 toxin, as determined by bis-benzimide staining and DNA ladder assays. However, IL-13 did not induce cell death. Flow cytometric analysis suggested that these cancer cell lines increased the sub-G1/G0 phase DNA population in a dose- and time-dependent manner (ranged between 10 and 30%) after treatment with IL-13 toxin. By Western blot analysis, cleavage of caspase-3 and PARP was observed after treatment with a high concentration of IL-13 toxin, also suggesting apoptotic cell death. In addition, the results of immunofluorescence and RT-PCR assays showed that the apoptosis-regulator, Bcl-2 was downregulated after treatment with IL-13 toxin, while Bax was upregulated. Moreover, significant nitrite production was detected in the HN12 cell line after treatment with IL-13 toxin for 48--96 h. Taken together, our results suggest that IL-13 toxin-induced cytotoxicity is at least partially mediated by the apoptosis and nitric oxide pathways. This information may be useful in developing specific approaches where apoptotic bodies from tumor cells may be used to pulse antigen-presenting cells for immunotherapy of cancer. PMID- 11862422 TI - An in vivo mouse reporter gene (human secreted alkaline phosphatase) model to monitor ovarian tumor growth and response to therapeutics. AB - PURPOSE: Developing new anticancer therapeutic regimens requires the measurement of tumor cell growth in response to treatment. This is often accomplished by injecting immunocompromised mice with cells from cancer tissue or cell lines. After treating the animals, tumor weight or volume is measured. Such methods are complicated by inaccuracies in measuring tumor mass and often animals must be killed to measure tumor burden. An in vivo tumor model system is presented in which the tumor cell line was stably transfected with a constitutively expressed marker gene: secreted human placental alkaline phosphatase protein (SEAP). The SEAP gene codes for a heat-stable protein that is produced at levels proportional to the amount of tumor cells in the animal. The SEAP protein is detectable in small blood samples so that animals can be repeatedly sampled over the trial period to monitor the course of tumor progression. METHODS: OCC1 ovarian carcinoma cells were stably transfected with pCMV-SEAP. The OCC1-SEAP cells were maintained in vitro to monitor the relationship between cell number and SEAP production. Experiments were performed in vivo to determine whether SEAP levels in blood corresponded to tumor burden. OCC1-SEAP cells were injected s.c. or intraperitoneally into nude mice and tumor volume was measured as well as plasma SEAP levels as the tumors developed. RESULTS: S.c. tumor volume correlated well with plasma SEAP levels ( R(2)=0.95). OCC1-SEAP cells were also injected intraperitoneally into nude mice and grown as abdominal tumors. After 3 weeks the animals were killed and the tumors were dissected and weighed. SEAP levels in plasma samples from the time of death correlated with intraperitoneal tumor weight ( R(2)=0.87). Experiments were performed to determine whether measuring SEAP levels could be used to monitor ovarian carcinoma cell response to platinum containing chemotherapeutic drugs. OCC1-SEAP cells cultured in vitro were treated with the platinum-containing drug carboplatin. Carboplatin treatment decreased both cell proliferation and SEAP levels in culture medium. The constitutive rate of SEAP secretion per cell (nanograms SEAP per microgram DNA) was found not to be altered by carboplatin treatment. Therefore changes in SEAP level reflect changes in OCC1 tumor cell number, and not changes in regulation of SEAP secretion due to platinum containing chemotherapeutic drug treatment. OCC1 cells were injected intraperitoneally into nude mice and the mice were treated with the platinum containing drugs cisplatin or carboplatin. Measurements of plasma SEAP over the treatment period showed OCC1-SEAP ovarian carcinoma growth to be inhibited by cisplatin and carboplatin treatment. CONCLUSION: The SEAP marker protein is constitutively expressed by tumor cells and blood levels are correlated with tumor cell number and burden. The results of these studies indicate that SEAP may be used as an in vivo reporter gene in a mouse model to monitor tumor growth and response to therapeutics. Future studies will utilize this model to investigate novel chemotherapeutic approaches to treating ovarian cancer. PMID- 11862423 TI - The effect of functional groups on reduction and activation of quinone bioreductive agents by DT-diaphorase. AB - PURPOSE: Bioreductive antitumor agents are an important class of anticancer drugs that include the clinically used drug, mitomycin C, and new agents such as EO9 and tirapazamine that have recently been tested in clinical trials. These agents require activation by reductive enzymes such as DT-diaphorase or NADPH:cytochrome P450 reductase. A major focus for improving cancer chemotherapy has been to increase the selectivity and targeting of antitumor drugs to tumor cells. Bioreductive antitumor agents are ideally suited to improving tumor selectivity by an enzyme-directed approach to tumor targeting. However, none of the bioreductive agents developed to date has been specific for activation by a single reductive enzyme. This is in part due to a lack of knowledge about structural factors that confer selectivity for activation by reductive enzymes. The purpose of this study was to investigate the ability of specific functional groups to modify reduction and activation of quinone bioreductive agents by DT diaphorase. METHODS: We used a series of model benzoquinone mustard (BM) bioreductive agents and compared the parent compound BM to MBM, which has a strong electron-donating methoxy group, MeBM, which has a weaker electron donating methyl group, CBM, which has an electron-withdrawing chloro group, and PBM and its structural isomer, meta-PBM (m-PBM), which both have sterically bulky benzene rings attached to the quinone moiety. We determined the rate of reduction of these agents by purified human DT-diaphorase under hypoxic and aerobic conditions. We also measured the cytotoxic activity of these agents in human tumor cell lines with and without the DT-diaphorase inhibitor, dicoumarol. RESULTS: Under hypoxic conditions in vitro, the t(1/2) values for reduction of the analogs by purified DT-diaphorase were 4, 6, 8, 9, 10 and 21 min for BM, MeBM, CBM, MBM, PBM and m-PBM, respectively. Under aerobic conditions the rank order of redox cycling after two-electron reduction by DT-diaphorase was MBM > MeBM > BM approximately CBM approximately PBM approximately m-PBM. The rate of reduction by DT-diaphorase of HBM, a non-alkylating analog of BM, was similar to that of BM under hypoxic conditions, and the rate of redox cycling under aerobic conditions was comparable to that of BM, suggesting that structural changes to the cytotoxic group of these BMs do not affect DT-diaphorase-mediated reduction and redox cycling potential. MBM, MeBM and PBM were more toxic than BM in the NCI H661 human non-small-cell lung cancer cells and SK-MEL-28 human melanoma cells, while CBM displayed significantly increased cytotoxic activity compared to BM only in the NCI H661 cells. m-PBM had similar cytotoxic activity compared with BM in both cell lines. These cell lines have moderate to high levels of DT diaphorase activity. When cells were pretreated with the DT-diaphorase inhibitor, dicoumarol, the cytotoxic activity of BM increased while that of MBM decreased in both cell lines, suggesting that BM was inactivated by DT-diaphorase while MBM was activated by this enzyme. Pretreatment of the SK-MEL-28 melanoma cells with dicoumarol resulted in an increased cytotoxic activity of MeBM, but pretreatment of the NCI-H661 cells did not affect the cytotoxicity of MeBM. This suggests, that similar to the results with BM, DT-diaphorase is an inactivating enzyme for MeBM in the SK-MEL-28 cell line. Dicoumarol had no significant effect on the cytotoxicity of CBM, PBM or m-PBM in both cell lines. CONCLUSIONS: These studies demonstrated that functional groups can significantly affect the reduction and activation of bioreductive agents by DT-diaphorase. All the functional groups decreased the rate of reduction of the quinone group by DT-diaphorase. Since MeBM and MBM, with electron-donating functional groups, and CBM with an electron withdrawing functional group had similar half-lives of reduction by DT diaphorase, steric rather than electronic effects of the functional groups appear to be more important for modifying the rate of reduction by DT-diaphorase. Steric effects on reduction by DT-diaphorase were also influenced by the position of the functional group on the quinone ring moiety, as the reduction of m-PBM was much slower than the reduction of PBM. The electron-donating methoxy and methyl functional groups increased the ability of the reduced products of MBM and MeBM to undergo redox cycling. DT-diaphorase appeared to be an activating enzyme for MBM. This may have resulted in part from increased formation of reactive oxygen species resulting from the increased redox cycling by MBM. In contrast, DT diaphorase was an inactivating enzyme for BM, and for MeBM in the SK-MEL-28 melanoma cells, possibly because the hydroquinone product of BM and MeBM may be less cytotoxic than the semiquinone produced by one-electron reduction by NADPH:cytochrome P450 reductase. PMID- 11862424 TI - Synergistic cytotoxic effect between serine-threonine phosphatase inhibitors and 5-fluorouracil: a novel concept for modulation of cytotoxic effect. AB - PURPOSE: The present study was undertaken to look for an agent or agents able to modulate the cytotoxic effect of 5-fluorouracil (FUra) and to investigate the role of serine-threonine phosphatase inhibitors on the cytotoxic effect of FUra. METHODS: The cytotoxicities of FUra and protein phosphatase inhibitors (PPIs) were evaluated by two different methods: a clonogenic assay and a proliferation assay. In the clonogenic assay, cancer cells were treated with various concentration of FUra with or without PPIs for 72 h. The drug-containing medium was replaced by fresh medium, the cultures incubated for an additional 10 days, and the colonies enumerated. In the proliferation assay the cells were treated with FUra alone or in combination with PPIs for 96 h and cytotoxicity was determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay based on the uptake of the tetrazolium dye. Thymidine kinase (TK) activity was determined based on the catalytic phosphorylation of [(3)H]d thymidine to [(3)H]dTMP. Incorporation of FUra into DNA and RNA was determined by treating the cells with [2-(14)C]fluorouracil for 72 h and measuring the radioactivity in the isolated DNA and RNA fractions. RESULTS: The serine threonine phosphatase inhibitors caliculin A (CAL), okadaic acid (OA) and microcystin-LR (MCLR) dose-dependently inhibited the growth of Clone 20 and Clone 5 cells of Colon 26 murine colon adenocarcinoma cells, human cervical cancer HeLa cells, human gastric cancer MKN 7 cells, and murine sarcoma S-180 cells in vitro. Among the compounds tested, MCLR at non-toxic concentrations was found to increase FUra incorporation into RNA and DNA in Clone 20 cells by 60% and 127%, respectively, to increase TK activity alone (twofold) as well as in combination with FUra (threefold), and to potentiate the cytotoxicity of FUra synergistically and cytospecifically in vitro. The cytotoxicity of FUra alone or in combination with MCLR, but not that of PPIs alone, was abrogated almost completely by exogenous thymidine (dThd), suggesting that inhibition of thymidylate synthetase (TS) is the growth-limiting event in the cytotoxic action of FUra even in combination with MCLR. CONCLUSIONS: The findings presented here suggest that MCLR synergistically and cytospecifically potentiates the antitumor activity of FUra with substantial improvement in the therapeutic index of FUra via enhancement of both DNA- and RNA-directed cytotoxicity. PMID- 11862425 TI - Entrapment by Cremophor EL decreases the absorption of paclitaxel from the gut. AB - BACKGROUND: Recent studies in mice and patients have shown that the low oral bioavailability of paclitaxel can be increased by coadministration of P glycoprotein blockers. However, in patients an increase in the oral paclitaxel dose from 60 to 300 mg/m(2) does not result in proportionally higher plasma levels. We hypothesized that the surfactant Cremophor EL, present in the formulation of paclitaxel, may be responsible for this nonlinear absorption by entrapping paclitaxel within the intestinal lumen, probably by inclusion in micelles. METHODS: Paclitaxel was administered to mdr1ab P-glycoprotein knockout mice with either the conventional (controls) or a seven-fold higher amount of Cremophor EL (test group). Plasma, gastrointestinal tissues with their contents and faeces were collected and analysed by high-performance liquid chromatography to determine the levels of paclitaxel and Cremophor EL. The critical micellar concentrations of Cremophor EL in the contents of the small intestine were also established by an in vitro assay. RESULTS: Paclitaxel recoveries in the faeces of the control and test groups were 7.6% and 35.8%, respectively. The peak plasma level and plasma AUC were reduced in the test group by about 75% and 40%, respectively. Only in mice from the test group did substantial quantities of paclitaxel together with Cremophor EL reach the caecum, thus passing through the small intestine. The concentration of Cremophor EL in the distal part of the small intestine and the caecum was 15 times higher in the test group and well above the critical micellar concentration of Cremophor EL. CONCLUSIONS: These results show that Cremophor EL prevents efficient uptake of paclitaxel from the gut, probably by entrapment within micelles. Other formulations should be developed for oral therapy with paclitaxel. PMID- 11862426 TI - Gender differences in the metabolism and pharmacokinetics of the experimental anticancer agent 5,6-dimethylxanthenone-4-acetic acid (DMXAA). AB - PURPOSES: Marked gender differences in the pharmacokinetics of many drugs have been reported. For the investigational anticancer drug, 5,6-dimethylxanthenone-4 acetic acid (DMXAA), negligible gender differences in the plasma pharmacokinetics have been observed in mice. The gender effects on the plasma pharmacokinetics of DMXAA were further investigated using the rat model. In addition, the in vitro metabolism and plasma protein binding of DMXAA in male and female mice, rats and humans were investigated. METHODS: DMXAA was administered to male and female rats by intravenous injection. DMXAA and its major metabolites formed in liver microsomes were determined by HPLC. Unbound DMXAA in plasma was separated by ultrafiltration followed by HPLC determination. RESULTS: In vivo kinetic studies indicated that female rats had 60%, 55% and 73% higher area under the plasma concentration-time curve (AUC) of DMXAA (2413 +/- 188 vs 1505 +/- 312 microM x h, P<0.05), elimination half-life (2.40 +/- 0.45 vs 1.55 +/- 0.33 h) and maximal plasma concentration (Cmax) (1236 +/- 569 vs 716 +/- 280 micro M), but 61% lower plasma clearance than male rats. In vitro studies indicated that male rats had a 67% higher glucuronidation activity (0.75 +/- 0.03 nmol/min per mg) than female rats (0.45 +/- 0.01 nmol/min per mg), resulting in a 96% faster intrinsic clearance (CL(int)) in the males than the females (6.36 +/- 0.65 vs 3.24 +/- .42 ml/min per g, P< 0.05). In contrast, female rats had 25% higher 6 methylhydroxylation activity (0.045 +/- 0.003 nmol/min per mg) than male rats (0.036 +/- 0.002 nmol/min per mg), resulting in a 57% faster intrinsic clearance (CL(int)) in the females than males (0.36 +/- 0.06 vs 0.23 +/- 0.05 ml/min per g). Overall, total CL(int) by both glucuronidation and 6-methylhydroxylation in male rats was 83% higher than in female rats (6.59 +/- 2.11 vs 3.60 +/- 1.07 nmol/min per g). Men ( n=4) had a significantly lower ( P<0.05) CL(int) for glucuronidation than women ( n=10), but a higher CL(int) for 6 methylhydroxylation, resulting in significantly higher total CL(int) in women than men (5.63 vs 8.33 nmol/min per g). There was no significant difference in either the total plasma protein or albumin concentration between male and female mice, rats or humans. CONCLUSION: There were significant gender-related differences in the metabolism and pharmacokinetics in the rat, in contrast to the mouse. This indicates a limited usefulness of the rat as a model for the study of DMXAA metabolism in relation to gender differences, although the gender differences in the in vitro metabolic capacity for DMXAA may provide an explanation for the gender differences in the pharmacokinetics in rats. Data from human liver microsomes may allow the prediction of gender effects in the in vivo pharmacokinetics of DMXAA. PMID- 11862427 TI - Peak plasma concentrations of doxorubicin in children with acute lymphoblastic leukemia or non-Hodgkin lymphoma. AB - PURPOSE: The peak plasma concentrations seem to play an important role in the toxicity of the anthracyclines. As there are only limited data in the literature about the distribution of doxorubicin in children, we assessed the peak plasma concentrations of doxorubicin in pediatric patients. PATIENTS AND METHODS: We collected 87 plasma samples at the end of infusion from 27 children with acute lymphoblastic leukemia (ALL) or non-Hodgkin lymphoma (NHL) treated with 30 mg/m(2) doxorubicin as a 1- or 2-h infusion once weekly for four weeks in the ALL BFM 95 or NHL-BFM 95 protocol. Plasma concentrations of doxorubicin were quantified by capillary electrophoresis, and the peak plasma levels for a uniform 2-h infusion were calculated. RESULTS: The geometric mean of all samples was 273 microg/l with a geometric coefficient of variation of 46.0%. This is in accordance with the peak plasma concentrations expected from simulations based on literature data from adults. High inter-individual as well as substantial intra individual variability was observed. Girls had slightly higher peak plasma levels than boys. Age, weight, and body mass index as well as laboratory parameters had no influence on the peak plasma concentrations. No cumulation of doxorubicin during therapy was observed. CONCLUSION: The peak plasma concentrations are similar in adults and children for both the absolute values as well as the variability; this indicates that there are no major differences in the distribution of doxorubicin in children and adults. PMID- 11862428 TI - Drug resistance in malignant rhabdoid tumor cell lines. AB - PURPOSE: We evaluated the in vitro sensitivity of four malignant rhabdoid tumor (MRT) cell lines to six chemotherapeutic agents: 5-fluororuacil, vincristine, carboplatin, doxorubicin, etoposide, and paclitaxel. We also sought to determine whether a defect in the p53 signaling pathway may contribute to the pronounced drug resistance of MRT. METHODS: MRT cells were treated with various concentrations of each drug and the effects on DNA synthesis were quantified using a thymidine incorporation assay. In addition, the effect of various concentrations of doxorubicin on cell growth was evaluated in all four cell lines. Functionality of the p53 pathway was evaluated by incubating cells with carboplatin or doxorubicin and monitoring the effects on the levels of the p53, p21(WAF1/CIP1), and MDM 2 proteins by Western blot analyses. RESULTS: Vincristine (EC(50) 0.5-2.9 n M) and doxorubicin (EC(50) 1.9-5.7 n M) were found to be most effective in inhibiting proliferation and were within clinically relevant concentrations. However, only doxorubicin exhibited cytotoxicity (EC(50) 2.4-13.1 n M), whereas vincristine and the other drugs tested were cytostatic. Interestingly, all four cell lines had remarkably similar dose response curves to all drugs tested, despite the fact that they were derived from different patients and arose in different tissues. When challenged with DNA-damaging drugs, p53 and the downstream effectors, p21(WAF1/CIP1) and MDM 2 were upregulated. CONCLUSIONS: These studies indicate that the p53 pathway is functional and responsive to DNA damaging drugs, and does not likely contribute to the drug resistance of MRT. The in vitro sensitivity of MRT cells to doxorubicin suggests that it may be a clinically important agent for the treatment of MRT. PMID- 11862430 TI - Phase I and pharmacokinetic study of 5-fluorouracil administered by 5-day continuous infusion in patients with hepatocellular carcinoma. AB - PURPOSE: In this study the maximum tolerated dose of 5-fluorouracil administered by 5-day (120-h) continuous infusion every 4 weeks was investigated and the pharmacokinetics in patients with hepatocellular carcinoma were evaluated. METHODS: Patients with hepatocellular carcinoma no longer amenable to established forms of treatment were eligible for the study. The starting dose of 5 fluorouracil was 300 mg/m(2) per day and doses were escalated in 50 mg/m(2) per day increments in successive cohorts of three new patients if tolerated. Pharmacokinetic studies were performed at the time of the first course of therapy. RESULTS: Enrolled in the study were 20 patients. The maximum tolerated dose was 500 mg/m(2) per day and the dose-limiting toxicity was stomatitis. Other toxicities were mild and well tolerated. Age, gender and associated liver cirrhosis were significant factors influencing 5-fluorouracil clearance. With regard to biochemical parameters, serum alanine aminotransferase and cholesterol levels were correlated with 5-fluorouracil clearance. CONCLUSIONS: The maximum tolerated dose for 5-day continuous infusion of 5-fluorouracil in hepatocellular carcinoma patients was 500 mg/m(2) per day. The recommended dose for phase II studies using this schedule is 450 mg/m(2) per day. Furthermore, the pharmacokinetic data obtained in this study may be useful in determining chemotherapy dosage adjustments for reduction of toxicity. PMID- 11862429 TI - PEG-asparaginase (Oncaspar) 2500 U/m(2) BSA in reinduction and relapse treatment in the ALL/NHL-BFM protocols. AB - PURPOSE: As previous data had shown that only two-thirds of patients had the predicted activity time courses when PEG-asparaginase 1000 U/m(2) was used in reinduction after native E. coli asparaginase in induction treatment of acute lymphoblastic leukaemia (ALL), drug monitoring was performed with the use of a higher dose. METHODS: Because one-third of patients had insufficient serum asparaginase activity time courses after a single dose of 1000 U/m(2) PEG asparaginase during reinduction treatment, a dose of 2500 U/m(2) PEG asparaginase, which is the approved dosage in Germany, was used in 39 reinduction and 20 relapse patients to determine whether prolongation of the activity time course may be possible with this higher dose, and to look for significant differences between reinduction and relapse patients. RESULTS: After 1, 2 and 3 weeks, the mean activities were 1113 +/- 699 U/l, 231 +/- 259 U/l, and 13 +/- 35 U/l in the reinduction patients, and 1078 +/- 649 U/l, 165 +/- 195 U/l and 19 +/- 28 U/l in the relapse patients, respectively. There were a considerable number of patients with a substantially shortened activity time course in both groups. In 10 of 39 reinduction patients and in 7 of 24 doses during relapse treatment, only activities <100 U/l were found after 1 week with a further fast decline. No statistically significant differences between the two patient groups could be shown at any time-point. CONCLUSIONS: Comparison of these data with activities after 1000 U/m(2) PEG-asparaginase showed no prolongation of the time with activity in the therapeutic range with the higher dose. Therefore, for a longer duration of therapeutic activity, administration of further doses is mandatory. PMID- 11862431 TI - Recombinant human interleukin-11 improved carboplatin-induced thrombocytopenia without affecting antitumor activities in mice bearing Lewis lung carcinoma cells. AB - PURPOSE: Interleukin-11 (IL-11) is a stromal cell derived multifunctional cytokine, which plays important roles in the hematopoietic and nonhematopoietic systems. Recombinant human IL-11 (rhIL-11) is used in the treatment of chemotherapy-induced thrombocytopenia. We have investigated the effects of rhIL 11 on the antitumor activity of chemotherapeutic agents and on thrombocytopenia in myelosuppressed mice bearing tumor cells. METHODS: We tested the effect of rhIL-11 on Lewis lung carcinoma (LLC) cell proliferation when used alone or in combination with three antitumor agents in vitro. Also, a newly developed chemotherapy-induced myelosuppressed mice model bearing LLC cells was used to study the effects of rhIL-11 on the antitumor activity and on thrombocytopenia. RESULTS: On its own, rhIL-11 (1-100 ng/ml) did not stimulate cell proliferation, and did not alter the antitumor activities of carboplatin, mitomycin C, or etoposide in vitro. In mice implanted with LLC cells (1 x 10(4)), carboplatin (50 mg/kg/day for 2 consecutive days, i.p.) inhibited tumor growth and caused thrombocytopenia. Treatment with rhIL-11 (500 microg/kg/day, from the day following the last dosing with carboplatin for 14 days, s.c.) successfully prevented thrombocytopenia without affecting the antitumor activity of carboplatin. With rhIL-11 there was no obvious effect on the red blood cell count, white blood cell count, or body weight. CONCLUSION: These results support the assertion that rhIL-11 may be a significant therapeutic agent for thrombocytopenia following cancer chemotherapy, and that it need be associated with little fear of tumor proliferation. PMID- 11862432 TI - Comparison of apoptotic, necrotic and clonogenic cell death and inhibition of cell growth following camptothecin and X-radiation treatment in a human melanoma and a human fibroblast cell line. AB - PURPOSE: We evaluated apoptotic, necrotic and clonogenic cell death and inhibition of cell growth in a human melanoma cell line (Sk-Mel-3) and a normal human fibroblast cell line (AG1522) following treatment with camptothecin (CPT) or with concurrent CPT and X-radiation. MATERIALS AND METHODS: Apoptotic and necrotic cell death was determined morphologically by dual-staining (propidium iodide, acridine orange). Inhibition of cell growth was determined from the number of cells remaining in the culture dish following treatment. RESULTS: In Sk Mel-3 cells: (a) after treatment with CPT alone, both apoptotic and necrotic cell death increased significantly ( P<0.05) relative to untreated controls; (b) after concurrent CPT and radiation treatment, however, only the increase in necrotic cell death was significant ( P<0.05) relative to cells receiving radiation alone; and (c) all assays of cellular effects/cytotoxicities were consistent in showing that CPT, given alone or with radiation, led to a substantial increase in cell kill. In contrast, in AG1522 cells: (a) there were no significant increases in apoptotic or necrotic cell death following either CPT alone or concurrent CPT and radiation; and (b) the clonogenic assay measured substantially higher cytotoxicities than the other assays. CONCLUSIONS: Necrotic cell death was more important than apoptotic cell death during concurrent CPT and radiation treatment in Sk-Mel-3 cells, but not in AG1522 cells. PMID- 11862434 TI - Antenatal diagnosis of spinal lipomas. AB - OBJECT: The purpose of this study was to highlight the key prenatal diagnostic features of fetal spinal lipomas. METHODS AND RESULTS: All fetuses in whom a spinal lipoma was suspected prenatally or diagnosed postnatally were included in the study. Lumbosacral lipoma was diagnosed in 16 fetuses (group 1), but at birth 3 of these were found to have other forms of occult dysraphism. In 2 other fetuses (group 2) a meningocele was suspected but at birth these lesions were found to be lumbosacral lipomas. The lipomas were covering large meningoceles in 13 fetuses and filling the cul-de-sac in 2 others. Three sets of parents decided on termination. Of the 13 children born with a spinal lipoma, 9 were operated on and 4 were managed conservatively. At the last follow-up, 7 were neurologically intact. CONCLUSION: The distinction between meningomyelocele and spinal lipoma is possible antenatally. The differential diagnosis between lipoma and the other forms of occult dysraphism is more difficult. PMID- 11862435 TI - NGF, NT-3 and Trk C mRNAs, but not TrkA mRNA, are upregulated in the paraventricular structures in experimental hydrocephalus. AB - OBJECTS: This study was designed to detect possible alterations in the expression of neurotrophins and trks in kaolin-induced hydrocephalus by in situ hybridization. METHODS AND RESULTS: Sixteen rats were treated by injection of 25 mg kaolin suspended in 0.1 ml of physiological saline into the cisterna magna. Four rats were injected with saline and served as controls. The kaolin-treated rats were divided into two groups studied 1 and 4 weeks after treatment. Rats were anesthetized and killed, and their brains were rapidly dissected and frozen. DNA oligonucleotide probes for nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3), and trkA, trkB, and C were labeled with [(35)S]dATP using terminal deoxyribonucleotidyl transferase for in situ hybridization. Hydrocephalic brains were also classified according to the degree of ventricular enlargement. The results observed were as follows. (1) The medial septal and striatal NGF mRNA levels increased with severity in animals. (2) Hippocampal trkB and BDNF mRNA levels increased with time in animals with moderate ventricular enlargement. (3) Expression of hippocampal trkB, trkC, and NT-3 mRNA increased in animals with moderate ventricular enlargement, while it apparently decreased in the large ventricular enlargement group reaching normal ranges. (4) In the corpus callosum there was an apparent increase in NGF, NT-3 and trkC mRNA, but not in trkA, in hydrocephalic animals. NT-3 EIA confirmed the presence of NT-3 protein increases in corpus callosum. It is therefore possible that simultaneous NGF, NT-3, and trkC receptor upregulation occurred in glial elements of the white matter. CONCLUSIONS: These results demonstrate that neurotrophins and their receptors are overexpressed in many damaged structures of the severely hydrocephalic brain. There were discrepancies in the distribution of NGF and trkA mRNA, and we hypothesize that NGF mRNA in the damaged white matter structure might be due to the reduced availability of other receptors, such as the low-affinity NGF receptors. PMID- 11862436 TI - Endoscopic treatment of suprasellar and third ventricle-related arachnoid cysts. AB - OBJECTIVE: To evaluate the role of neuroendoscopy in the treatment of rare suprasellar and other third ventricle-related arachnoid cysts. METHODS: A review of supratentorial midline arachnoid cysts treated by endoscopic fenestration. RESULTS: Ten cases were identified. The median age of these ten patients was 7.5 years. In three a cysto- or ventriculoperitoneal shunt had previously been inserted. Six cysts were located in the suprasellar/prepontine area, one was a combined suprasellar/middle fossa cyst, and three were complex cysts arising mainly in the quadrigeminal cistern and extending into the third ventricle. Successful endoscopic fenestration to both the ventricular system and the basal cisterns ("bipolar" fenestration) was achieved in all cases. In seven cases a cystoventriculostomy and cystocisternostomy was performed, while in three a cystoventriculostomy was combined with a third ventriculostomy, achieving additional communication to the subarachnoid space. The previously inserted shunts were removed at the end of the procedure. The median follow-up period was 17 (range 1-30) months. The clinical outcome was excellent in all cases, despite only marginal reduction in the cyst size and persistence of ventriculomegaly in some cases. No further treatment to the cyst was required during the period of follow-up. CONCLUSION: Third ventricle-related arachnoid cysts can be satisfactorily and safely treated by endoscopy. This approach leaves the patient shunt independent. PMID- 11862437 TI - Neurosurgical treatment of meningiomas in children and young adults. AB - OBJECT: We studied the frequency, functional outcome, association with neurofibromatosis (NF) and relapse in patients operated on for meningiomas at age 0-20 years in three Norwegian centers between 1972 and 1999. METHODS: Information was collected by examining case notes and histology records, conducting telephone interviews and performing new radiological investigations. Twenty-seven patients were identified. Five had NF, and all 5 developed multiple tumors. In non-NF patients, relapse occurred in 2 out of 19 who underwent total tumor resection, and in all 3 whose surgery was non-radical. There was 1 postsurgical death, and no other major complications. Most patients had few complaints or none at all resulting from their previous tumor or treatment. CONCLUSIONS: Meningiomas without NF have a good prognosis in children and young adults if the tumor can be removed radically. However, relapse can occur many years after the primary operation. PMID- 11862438 TI - Intracystic chemotherapy with bleomycin in the treatment of craniopharyngiomas. AB - OBJECT: Since bleomycin has not yet been used very frequently in the treatment of patients with craniopharyngioma, it seemed important to document the course of a series of such patients treated with this preparation. METHODS AND RESULTS: Local chemotherapy with bleomycin was performed in 24 patients (20 children and 4 adults), 16 of whom presented with cystic or mixed (solid/cystic) craniopharyngioma and 8, with recurrent cystic craniopharyngioma. The drug was administered through an Ommaya reservoir, which was placed either by using a direct surgical approach (6 patients) or a stereotactic approach (16 patients), or with endoscopic assistance in patients with hydrocephaly (2 patients). Injection of bleomycin was always preceded by a water-tightness test. Each patient received a 3-mg dose of bleomycin every other day. The total dose of bleomycin injected ranged from 28 mg to 150 mg. Most patients (17, or 70%) were treated only with intracystic chemotherapy. Chemotherapy was followed by surgery in 7 patients. Five were operated on at the beginning of our study, and 2 required surgery because chemotherapy yielded poor results. A toxic dose was injected in 1 patient only: a severe complication, i.e. blindness, was observed. The follow-up period ranged from 2 years to 10 years. CONCLUSION: Our results show that bleomycin can be an alternative in the treatment of cystic craniopharyngiomas or cystic recurrences, as it reduces surgical morbidity and improves clinical results. PMID- 11862439 TI - White matter disease in children treated for malignant brain tumors. AB - OBJECTS: The objects of the study reported were to recognize different patterns of white matter disease (WMD) in the follow-up of children after surgery, radiation and/or chemotherapy for malignant primary brain tumors and to evaluate statistical data on the incidence of WMD and various risk factors. METHODS: Magnetic resonance imaging (MRI) records were evaluated retrospectively in the routine follow-up (range 6 months to 15 years after surgery) of 44 children with malignant primary brain tumors treated with surgery and radiotherapy and/or chemotherapy. RESULTS: WMD was diagnosed in 28 children and subclassified into circumscribed white matter lesions (WML) and diffuse atrophy. WML were the most common finding ( n=13), followed by atrophy ( n=7) and the combination of both ( n=8). Statistical analysis revealed slightly more frequent atrophy in children younger than 5 years. WML could be linked with supratentorial location of the tumor, follow-up longer than 5 years, and the presence of a ventricular shunt. Intrathecal chemotherapy was also a factor, but because of the small sample size of the group this might not be valid. None of the children had neurological deficits attributed to these findings, but the impact on neuropsychological development was not determined. PMID- 11862440 TI - Prenatal diagnosis of suprasellar arachnoid cyst and postnatal endoscopic treatment. AB - Arachnoid cysts are benign developmental anomalies that occur in the cerebrospinal axis in relation to the arachnoid membrane. An antenatal ultrasound scan first raises the suspicion of arachnoid cysts, but misdiagnoses have been reported. Confirmatory antenatal magnetic resonance imaging (MRI) is very useful to delineate anatomical detail and help in correct diagnosis. This helps proper counselling and treatment planning. Controversy also surrounds the surgical management, in terms of both the indications and the procedures employed. We report the successful endoscopic treatment, in a neonate, of an arachnoid cyst diagnosed by antenatal ultrasound and MRI. In this case accurate diagnosis and detailed assessment led to objective counselling and helped to optimise neonatal management. PMID- 11862441 TI - Growth rate of cerebral hydatid cyst, with a review of the literature. AB - This extremely rare case was one of secondary solitary cerebral echinococcosis associated with possible cerebral thromboembolism. A 7-year-old girl living in a rural area was admitted to our hospital with a history of headache, right-sided hemiparesis, and dysphasia. She had been treated 6 months previously for a cerebral infarct, diagnosed from sudden altered consciousness and a myoclonic generalised convulsion. The growth rate determined for the cerebral hydatid cyst was about 4.5 cm during the 6-month period. In children a parasitic cyst can be the source of a cerebral embolus, particularly in areas where hydatid disease resulting from cardiac echinococcosis is endemic. PMID- 11862442 TI - Supratentorial hydatid cyst with cerebellar signs: a rare case of diaschisis. AB - BACKGROUND: Intracranial hydatid disease has a distinct predominance in the pediatric age group and still causes serious problems in endemic areas. CASE REPORT: A 7-year-old girl admitted with a 3-month history of illness involving the main symptoms of ataxic gait, apraxia, headache, and tremor and with positive cerebellar signs and papilledema is presented. RESULTS: Cranial computerized tomography and magnetic resonance imaging revealed a right temporoparietal spherical lesion measuring 50 x 60 x 80 mm, which had a significant mass effect. A preoperative diagnosis of intracranial hydatid cyst was confirmed during the surgical procedure, which allowed removal of the cyst intact. The postoperative course was uneventful. CONCLUSIONS: The aim of this presentation is to emphasize the necessity for considering diaschisis--inhibition of function produced by a focal disturbance in a portion of the brain at some distance from the original site of injury, but anatomically connected with it through fiber tracts. We believe that this case, with a mass lesion in a temporoparietal location and definite clinical manifestations with plentiful cerebellar signs, is a good example of this rare phenomenon. PMID- 11862443 TI - Ventricular diverticulum at the posterior horn of the lateral ventricle presenting as aplasia cutis congenita. AB - Non-communicating hydrocephalus can cause rupture of the lateral or third ventricle, generally in the medial wall of the trigone or the posterior wall of the third ventricle, resulting in a cystic lesion known as ventricular diverticulum. In this paper, we describe a rare case of ventricular diverticulum located in the posterior horn of the lateral ventricle and expanding to bulge with a convexity reaching into the subdural space in a neonate. Aplasia cutis congenita and a bone defect were also present in the same neonate. Early surgical repair of the scalp defect and ventriculoperitoneal shunting were performed. In addition to illustrating the rare co-existence of a ventricular diverticulum at the posterior horn of the lateral ventricle and aplasia cutis congenita in a neonate, this case also offers new insights into the pathogenesis of these congenital anomalies. PMID- 11862444 TI - Acute paraplegia revealing an intraspinal neurenteric cyst in a child. AB - The authors report a case of intraspinal neurenteric cyst in a 22-month-old child, who presented with acute paraplegia following a vesicourethrogram. Despite 8 days' delay in surgical decompression, he made a complete neurological recovery. Neurenteric cysts are rare congenital lesions of the spinal canal lined with an epithelium of endodermal origin. They are usually located at the cervicothoracic junction and present with progressive mild to moderate signs of myelopathy. This is a unique case in regard both to its clinical presentation and to the excellent outcome after 8 days of complete paraplegia. PMID- 11862446 TI - 17th meeting of the Spanish Society for Pediatric Neurosurgery (SENEP), Santander, Spain, 14-15 December, 2000. PMID- 11862445 TI - Intramedullary immature teratoma in a young infant involving a long segment of the spinal cord. AB - Intramedullary teratoma is a rare lesion, located in the majority of cases in the lumbosacral area, and such lesions involving an extensive area of the spinal cord in young infants have seldom been reported. We present the case of a 3-month-old girl with an intramedullary spinal immature teratoma extending from C-5 to T-12, which was totally removed. The patient had suffered from paraplegia for 15 days, after which spinal MRI revealed a heterogeneously enhancing intramedullary lesion. Biopsy of the lesion demonstrated mature intestinal tissue. After total removal of the tumor, paraplegia and sphincter disturbances improved. Intramedullary teratoma should be included in the differential diagnosis of holocord tumors in young infants with rapidly progressing symptoms and if found should be radically excised. This case also emphasizes the importance of histological diagnosis and demonstrates the possibility of neurological recovery even in the case of paraplegia lasting for more than 1 month in a young infant. PMID- 11862449 TI - Immunohistochemical and ultrastructural study of basophilic inclusions in adult onset motor neuron disease. PMID- 11862451 TI - The actin loci in the genus Drosophila: establishment of chromosomal homologies among five palearctic species of the Drosophila obscura group by in situ hybridization. AB - Chromosomal homologies among the four palearctic Drosophila obscura group species D. ambigua, D. tristis, D. obscura, and D. subsilvestris and the "trans palearctic" species D. bifasciata were established by in situ hybridization using the 5C actin gene of D. melanogaster as a probe. In all species two labeling sites were detected in each of chromosomal elements C and E and one in each of chromosomal elements A and D. In addition one labeling site was detected on element B for the species D. subsilvestris and D. bifasciata. The conservative distribution pattern of the genes of the actin multigene family, the similarities of the locations of the actin genes in the chromosomes of the five species studied, together with the concordant evidence of synteny of visible and other genetic markers as well as the similarities in banding patterns, all agree with the conclusion that the chromosomal elements have retained their essential identity throughout the evolution of these species. Using in situ hybridization detailed information of some homologous regions of chromosomes can also be established. PMID- 11862452 TI - The genomic complexity of micro B chromosomes of Brachycome dichromosomatica. AB - A major sequence component of the micro B chromosome of Brachycome dichromosomatica (2 n=4) is the tandem repeat Bdm29, which was found by in situ hybridisation to be distributed along the entire length of the chromosome. A high copy number of this sequence does not occur as a regular feature of the A chromosomes in this species but it was found in infrequent individuals in two wild populations that were analysed. In these instances Bdm29 is localised within heterochromatic, polymorphic segments on the long arm of chromosome 1. The origin of the micro B chromosomes was investigated by determining whether they are related to this A chromosome polymorphism by simple excision and/or integration. Results obtained by using Bdm29, together with a newly isolated repeat sequence, Bdm54, and a number of other sequences known to occur on the micro B chromosome, as probes in in situ hybridisation and Southern analysis demonstrated that the formation of micro B chromosomes is a complex multistep process. The observation that the genomic organisation of the micro B chromosome is unlike anything found on the A chromosomes precludes their origin by simple excision and also indicates that micro Bs do not integrate directly into the A complement to form polymorphic heterochomatic segments. PMID- 11862453 TI - Non-isotopic mapping of ribosomal RNA synthesis and processing in the nucleolus. AB - The precise location of ribosomal RNA (rRNA) synthesis within the nucleolus is the subject of recent controversy; some investigators have detected nascent RNA in the dense fibrillar components (DFCs) while others have localized transcription to the fibrillar centers (FCs). We endeavored to resolve this controversy by applying a new technique for non-isotopic labeling of RNA and examined the synthesis and movement of non-isotopically labeled rRNA within the nucleolus. We found that rRNA is synthesized only in a restricted area of DFCs, also involving the boundary region with FCs. We traced a movement of RNA from transcription sites through DFCs to granular components. Our results indicate functional compartmentalization of DFCs with respect to the synthesis and processing of precursor rRNA. In situ mapping of the 5' leader sequence of the 5' external transcribed spacer together with transcription labeling indicated that transcription and the first steps in processing of precursor rRNA are spatially separated. Surprisingly, the results also pointed to a partially extended conformation of newly synthesized precursor rRNA transcripts. PMID- 11862454 TI - Disassembly of nuclear bodies during arousal from hibernation: an in vitro study. AB - In previous studies we demonstrated that during hibernation cell nuclei contain structural constituents usually absent in euthermia. The rapid disappearance of such nuclear bodies upon arousal makes very difficult the in vivo investigation of the disassembly process, which could clarify their functions in nuclear metabolism in the hibernator. In the present study we subjected liver samples taken from hibernating edible dormice ( Glis glis) to different in vitro experimental conditions: at 4 degrees C, to preserve the hypothermic state of the hibernating organism; at 37 degrees C, to simulate the drastic increase in body temperature occurring during arousal; at 37 degrees C, in culture medium containing 10(-5) M delta opioid D-Ala2- D-Leu5 enkephalin, which mimics the activity of the hibernation induction trigger in hibernators. Electron microscopic analysis of hepatocyte nuclei at increasing incubation times revealed the subsequent steps of disassembly of coiled bodies, amorphous bodies and fibro granular material, the unusual structural constituents accumulating during hibernation in these nuclei. We demonstrated that: (1) a temperature of 37 degrees C induces the disappearance of all nuclear bodies typical of hibernation in a few minutes; (2) both low temperature and hibernation-triggering opioid are able to slow down, although to different extents, the process of disassembly of nuclear bodies; (3) the fibro-granular material rapidly disappears during the early phases of incubation; while (4) coiled bodies and amorphous bodies progressively disassemble as fibrous material. Our results support previous hypotheses based on in vivo observations about a possible role for coiled bodies, amorphous bodies and fibro-granular material as storage/assembly sites of molecules needed for the rapid and massive resumption of transcriptional and post transcriptional activities upon arousal and suggest a strict correlation between the dynamics and metabolic rate of nuclear bodies. PMID- 11862455 TI - Colchicine promotes a change in chromosome structure without loss of sister chromatid cohesion in prometaphase I-arrested bivalents. AB - In somatic cells colchicine promotes the arrest of cell division at prometaphase, and chromosomes show a sequential loss of sister chromatid arm and centromere cohesion. In this study we used colchicine to analyse possible changes in chromosome structure and sister chromatid cohesion in prometaphase I-arrested bivalents of the katydid Pycnogaster cucullata. After silver staining we observed that in colchicine-arrested prometaphase I bivalents, and in contrast to what was found in control bivalents, sister kinetochores appeared individualised and sister chromatid axes were completely separated all along their length. However, this change in chromosome structure occurred without loss of sister chromatid arm cohesion. We also employed the MPM-2 monoclonal antibody against mitotic phosphoproteins on control and colchicine-treated spermatocytes. In control metaphase I bivalents this antibody labelled the tightly associated sister kinetochores and the interchromatid domain. By contrast, in colchicine-treated prometaphase I bivalents individualised sister kinetochores appeared labelled, but the interchromatid domain did not show labelling. These results support the notion that MPM-2 phosphoproteins, probably DNA topoisomerase IIalpha, located in the interchromatid domain act as "chromosomal staples" associating sister chromatid axes in metaphase I bivalents. The disappearance of these chromosomal staples would induce a change in chromosome structure, as reflected by the separation of sister kinetochores and sister axes, but without a concomitant loss of sister chromatid cohesion. PMID- 11862456 TI - Electron microscope mapping of the pericentric and intercalary heterochromatic regions of the polytene chromosomes of the mutant Suppressor of underreplication in Drosophila melanogaster. AB - Breaks and ectopic contacts in the heterochromatic regions of Drosophila melanogaster polytene chromosomes are the manifestations of the cytological effects of DNA underreplication. Their appearance makes these regions difficult to map. The Su(UR)ES gene, which controls the phenomenon, has been described recently. Mutation of this locus gives rise to new blocks of material in the pericentric heterochromatic regions and causes the disappearance of breaks and ectopic contacts in the intercalary heterochromatic regions, thereby making the banding pattern distinct and providing better opportunities for mapping of the heterochromatic regions in polytene chromosomes. Here, we present the results of an electron microscope study of the heterochromatic regions. In the wild-type salivary glands, the pericentric regions correspond to the beta-heterochromatin and do not show the banding pattern. The most conspicuous cytological effect of the Su(UR)ES mutation is the formation of a large banded chromosome fragment comprising at least 25 bands at the site where the 3L and 3R proximal arms connect. In the other pericentric regions, 20CF, 40BF and 41BC, 15, 12 and 9 new bands were revealed, respectively. A large block of densely packed material appears in the most proximal part of the fourth chromosome. An electron microscope analysis of 26 polytene chromosome regions showing the characteristic features of intercalary heterochromatin was also performed. Suppression of DNA underreplication in the mutant transforms the bands with weak spots into large single bands. PMID- 11862457 TI - Lymphoblasts already in the DNA synthesis phase of the cell cycle can be reversibly arrested at the R/G transition. AB - Early and late S-phase of the cell cycle are separated by the R-band/G-band (R/G) transition. This corresponds to the time at which R-band synthesis has been completed while G-band synthesis has yet to begin. The aim of this work was to study cell cycle kinetics during S-phase using different blocking agents: mimosine, methotrexate, 5-fluorouracil, 5-fluoro-2'-deoxyuridine and an excess of thymidine. The stage at which these blocking agents arrest the cell cycle and their efficiency at blocking Epstein-Barr virus transformed lymphoblasts at the R/G transition were evaluated using flow cytometric techniques. Mimosine blocked 90% of the cells near the G1/S-phase boundary. Methotrexate, 5-fluoro-2' deoxyuridine and 5-fluorouracil, and particularly thymidine, let a significant proportion of cells enter S-phase. The cells were released from the arrest state and their progression through early S-phase was monitored by flow cytometry. Before the cells reached the R/G transition, a second agent was added to inhibit cell cycle progression. For example, the use of mimosine followed by thymidine allowed up to 60% of the cells to be blocked at the R/G transition. The arrest of DNA replication at the R/G transition was confirmed by a marked decrease of 5 bromo-2'-deoxyuridine (BrdUrd) incorporation, revealed by using bivariate flow cytometric analysis. The blocking agent was then removed and the cell cohort was released in the presence of BrdUrd so that replication banding analysis could be performed on the harvested mitotic cells. This yielded a mitotic index of approximately 10% and chromosomes showing replication bands. Flow cytometric analysis combined with cytogenetic banding analysis suggested that the R/G transition is an arrest point within the S-phase of the cell cycle and allowed us to conclude that only cells that have already initiated S-phase are blocked at this point. It corresponds to a susceptible site where S-phase can be arrested easily. The R/G transition could also be a regulatory checkpoint within S-phase, a checkpoint that could respond to imbalance in deoxyribonucleotide pools. PMID- 11862458 TI - Genomic imprinting of a placental lactogen gene in Peromyscus. AB - The mammalian genome contains over 30 genes whose expression is dependent upon their parent-of-origin. Of these imprinted genes the majority are involved in regulating the rate of fetal growth. In this report we show that in the deer mouse Peromyscusthe placental lactogen-1-variant ( pPl1-v) gene is paternally expressed throughout fetal development, whereas the linked and closely related pPl1gene is expressed in a biallelic manner. Neither the more distantly related pPl2Agene, nor the Mus Pl1gene displays any preferential expression of the paternal allele, suggesting that the acquisition of imprinting of pPl1-v is a relatively recent event in evolution. Although pPl1 expression is temporally mis regulated in the dysplastic placentae of hybrids between two Peromyscus species, its over-expression cannot account for the aberrant phenotypes of these placentae. We argue that the species-specific imprinting of pPl1-v, encoding a growth factor that regulates nutrient transfer from mothers to their offspring, is consistent with the parent-offspring conflict model that has been proposed to explain the evolution of genomic imprinting. PMID- 11862459 TI - Structure and expression of an Otx5-related gene in the dogfish Scyliorhinus canicula: evidence for a conserved role of Otx5 and Crxgenes in the specification of photoreceptors. AB - We report the full-length coding sequence and the expression pattern during neurulation and early organogenesis of ScOtx5, a novel member of the Otx gene family in the dogfish Scyliorhinus canicula. Phylogenetic analyses confirm that ScOtx5 is closely related to the Xenopus XlOtx5/ 5bgenes, and also to the Crx genes characterized in mammals and zebrafish. This supports the hypothesis that these genes define a third gnathostome Otx orthology class. During neurulation, ScOtx5 transcripts are detected in the foregut diverticulum and the anterior neuroectoderm. At the onset of organogenesis, ScOtx5 is transcribed over a broad domain spanning the whole prosencephalon and mesencephalon, albeit with a much lower signal intensity than its paralogues Otx1 and Otx2. At later stages, four major expression sites are observed: the developing eye and epiphysis, the olfactory placodes and a broad epidermal domain in the dorsal part of the head. In the embryonic eye, the signal is first detected in the presumptive pigmented retina and slightly later in the adjacent outer layer of the neural retina, fated to photoreceptors. The comparison of this expression pattern with those of osteichthyan Otx genes suggests that a role in the specification of photoreceptors may correspond to a functional specialization of Otx5and Crx genes, fixed early in the gnathostome lineage, prior to the splitting of chondrichthyans and osteichthyans. In contrast, the roles played by ScOtx5 in the retinal pigmented epithelium or in the olfactory placodes may be fulfilled by different combinations of paralogous genes in other gnathostome taxa. PMID- 11862460 TI - Expression and function of the LIM homeodomain protein Apterous during embryonic brain development of Drosophila. AB - We analyzed the expression and function of the LIM-homeodomain transcription factor Apterous (Ap ) in embryonic brain development of Drosophila. Expression of Ap in the embryonic brain begins at early stage 12 and is subsequently found in approximately 200 protocerebral neurons and in 4 deutocerebral neurons. Brain glia do not express Ap. Most of the Ap-expressing neurons are interneurons and project their axons across the midline to the contralateral hemisphere; a smaller subset projects their axons into the ventral nerve cord. A few Ap-expressing neurons project to the ring gland, suggesting that they are neurosecretory cells. In ap loss-of-function mutants, some of the protocerebral and deutocerebral interneurons that express Ap in the wild type show axon pathfinding errors and fasciculation defects in the brain, notably in the fascicles of the brain commissure. In contrast, the interneurons that project to the ring gland do not appear to be affected in ap mutants. Thus, in brain development, Ap is required for correct axon guidance and fasciculation of interneurons, and Ap-expressing cells may also be involved in the brain neuroendocrine system. PMID- 11862461 TI - Gdf5 is expressed in the developing skeleton of median fins of late-stage zebrafish, Danio rerio. AB - The dynamic expression of Gdf5 is described in the developing skeleton of the median fins of late-stage zebrafish, Danio rerio (6-45 days post-fertilization). In situ hybridization revealed expression in the mesenchyme between cartilage condensations of the endoskeletal supports of the dorsal, anal, and caudal fins. As development proceeds, the expression domains expand distally to surround tips of developing cartilages, consistent with a role in cartilage growth and differentiation. Gdf5 is later expressed in the segmenting regions of the dorsal and anal fin radials, which may indicate a role in segmentation. After growth to 7.5 mm, Gdf5 transcripts can no longer be detected in any of the median fins, nor is Gdf5 expression reinitiated in later development of the median fin skeleton. PMID- 11862462 TI - Expression pattern of novel chick T-box gene, Tbx20. AB - Little is known about the molecular mechanisms involved with the initial specifications of the cardiac mesoderm. In order to identify potential regulatory factors that play important roles in early heart specification, we attempted to isolate the chick H15-related T-box gene and analyze its expression pattern during early development. The chick Tbx20 gene was found to be highly homologous to human, mouse, and zebrafish hrT/Tbx20. Its expression was initially detected in the posterior lateral mesoderm, after which it expanded to the anterior and was intensively co-expressed with a cardiogenic gene, Nkx2.5, in the anterior lateral mesoderm. PMID- 11862463 TI - Distinct expression patterns of two zebrafish homologues of the human APP gene during embryonic development. AB - The human amyloid protein precursor (APP) gene correlates with early onset of Alzheimer's disease in humans. We have identified two APP homologues in zebrafish, which we call appa and appb. They show a high degree of identity to human APP particularly in the beta APP42 and the transmembrane domain. Widespread expression of both appa and appb was detected from mid-gastrulation until the bud stage. During segmentation, the two genes diverged in their pattern of expression: at 14 h post-fertilisation (hpf) and 18 hpf both genes were expressed rostrally in the prospective CNS, but only appa was found caudally in the paraxial segmental plate and presomitic mesoderm, excluding the midline. In contrast, appb was found caudally in the neural rod at 14 hpf and the developing spinal cord at 18 hpf. Later, at 24 hpf both genes shared common expression domains, namely the telencephalon, the ventral diencephalon, the trigeminal ganglia, and the posterior lateral line ganglia. Unique expression domains for appa were the lens, the otic vesicles and the somites, while appb was expressed in a serially repeated set of nuclei within the hindbrain, the ventral mesencephalon and the motoneurones of the developing spinal cord. PMID- 11862464 TI - Expression of the anti-dorsalizing morphogenetic protein gene in the zebrafish embryo. AB - The BMP3 related anti-dorsalizing morphogenetic protein (ADMP) has been proposed to function in the organizer of chick and Xenopus embryos. We report here the cloning and expression pattern of a zebrafish admp gene. The gene is expressed in involuting cells of the embryonic shield, but not in the non-involuting forerunner cells. During gastrulation, admp transcripts are detected in the posterior prechordal plate, in the notochord primordium and in cells of the dorsal blastoderm margin. Expression is also detectable in the neuroectoderm overlying the posterior prechordal plate. Expression persists in the tail bud until the end of somitogenesis while expression in other areas disappears during early somitogenesis stages. PMID- 11862466 TI - JunB negatively regulates AP-1 activity and cell proliferation of malignant mouse keratinocytes. AB - OBJECTIVE: Previously we have shown that a malignant mouse keratinocyte cell line, 10Gy5, has elevated AP-1 transactivation and reduced JunB protein levels compared to its parental benign cell line, 308, and that the tumorigenicity in the 10Gy5 cells could be blocked by a dominant negative c-Jun mutant protein. We wished to determine whether the change in JunB protein levels could account for the elevated AP-1 activity and whether re-expression of JunB in malignant 10Gy5 cells altered their proliferative capacity. DESIGN: In the current study, we reduced JunB expression in benign 308 cells with antisense oligonucleotides and increased JunB expression in malignant 10Gy5 cells by stable transfection of a JunB expression vector. RESULTS: Increased AP-1 activity was detected after treatment of the benign 308 cell line with JunB antisense oligonucleotides that reduced JunB protein levels. Stably JunB-transfected clones of malignant 10Gy5 cells showed decreased AP-1 activity, slowed in vitro cell proliferation and reduced tumor growth when xenografted to athymic nude mice. CONCLUSION: These findings suggest that expression of JunB protein has a negative effect on malignant tumor cell proliferation in part through its ability to inhibit AP-1 transactivation. PMID- 11862467 TI - The multicentric epitheloid hemangioendothelioma of bone: a case example and review of the literature. AB - Epitheloid hemangioendothelioma of bone is a rare primary bone malignancy. Diagnosis can be difficult for physicians who are not experienced with bone neoplasms. We also had difficulties in diagnosis and treatment in one of our patients with a multicentric epitheloid hemangioendothelioma involving the pelvis, left femur, and left tibia. We recommend a complete skeletal survey with magnetic resonance imaging because it can reveal previously undetected lesions on conventional radiographs. We performed an extraarticular resection of the hip joint including the spina iliaca anterior inferior, resection of the femur, and amputation of the lower leg. The femur was replaced by a modular endoprosthesis. The patient is provided with an above-knee prosthesis and is able to walk even longer distances with a cane 2 years after surgery. In our opinion it is necessary to perform a wide resection of this tumour in order to treat a patient with curative intention. Palliative radiotherapy should only be used for a non resectable tumour or in metastatic disease. Chemotherapy is not a treatment option. PMID- 11862468 TI - Comparison of processing four and five times the patients' blood volume during peripheral blood stem cell collection and analysis of CD34+38--and CD34+49d+ subsets during apheresis. AB - PURPOSE: We investigated whether increasing the patients' processed blood volume (BV) during peripheral blood stem cell collection (PBSCC) from four to five times leads to a greater yield of CD34+ cells. We also studied the kinetics of CD34+38- and CD34+49d+ subsets and compared the amount of transfused cells with engraftment. METHODS: All patients ( n=20) received chemotherapy followed by G CSF for PBSC mobilization. Samples from the patients' peripheral blood and the PBSC harvests were taken after processing 1-, 4-, and 5 times the patients' calculated BV. RESULTS: The mean total yields of CD34+, CD34+38-, and CD34+49d+ cells were 15.69-, 1.13- and 4.17 x 10(6)/kg body weight, respectively. The mean increase for these subsets between 4- and 5 BV was 10%, 8%, and 21%, respectively. Based on the mean number of 2.25 (range 2-3) planned courses of high-dose chemotherapy (HDC) per patient, the mean yield of CD34+ cells per kg body weight and intended course of HDC after 4- and 5 BV was 6.31- and 6.97 x 10(6) ( P=0.014). Twenty HDC were evaluable for engraftment. There was some correlation between the number of transfused CD34+ and CD34+38- cells and WBC engraftment ( r = -0.66 and --0.69; P<0.01) and CD34+ cells and platelet engraftment ( r = -0.56; P= 0.013). No toxicity occurred during PBSCC, although the mean platelet count dropped by 50% which must be kept in mind regarding the additional application of anti-coagulants and the fact that most patients had large indwelling catheters. CONCLUSION: Processing 4 BV is sufficient to collect >5 x 10(6) CD34+ cells/kg body weight and intended course of HDC in most patients, although extension to 5 BV further increases the total yield of CD34+ cells. PMID- 11862469 TI - Comparison of interleukin-12 with lung cancer and malignant lymphoma undergoing autologous peripheral blood stem cell transplantation. AB - PURPOSE: High-dose chemotherapy with peripheral blood stem cell transplantation (PBSCT) has become an important treatment for solid tumors including lung cancer. METHODS: We measured IL-12 levels in patients with lung cancer undergoing autologous PBSCT in order to elucidate the role of IL-12 in immune response recovery following stem cell transplantation. RESULTS: Compared to IL-12 levels at 1 week after PBSCT for lung cancer patients, those at 3 weeks were significantly increased ( P<0.01). In contrast, serum IL-12 levels in malignant lymphoma patients did not change significantly. There were no significant differences in levels of other cytokines between 1 week and 3 weeks after transplantation in patients with lung cancer. The frequency of helper/inducer T cells was increased in peripheral blood 1 week after transplantation in both lung cancer and malignant lymphoma patients. There was a significant increase in activated T cell numbers following PBSCT. Furthermore, high levels of other activated T cells persisted in the post-PBSCT period in patients with lung cancer and the number of cytotoxic T cells significantly increased. Natural killer (NK) cell numbers also tended to increase, although that of malignant lymphoma was not significant. A strong correlation was observed between serum IL-12 levels and NK cell numbers and interferon-gamma levels in lung cancer not but in malignant lymphoma patients. The analysis of transfused PBSC showed that the numbers of granulocyte/macrophage colony-forming units were similar in lung cancer and malignant lymphoma patients. However, the number of CD34+ cells was significantly higher in lung cancer than in malignant lymphoma patients. All of the CD34+ subpopulations were lower in percentage in patients with lung cancer than in patients with malignant lymphoma. In particular, the CD34+ CD33- subpopulation was significantly lower in percentage in lung cancer patients. CONCLUSION: Our findings suggest that PBSC in lung cancer are potent mediators of anticancer activity and that they might play an immunotherapeutic role against autologous malignant cells. PMID- 11862470 TI - Immunostimulatory effect of natural clinoptilolite as a possible mechanism of its antimetastatic ability. AB - PURPOSE: Many biochemical processes are closely related to ion exchange, adsorption, and catalysis. Zeolites reversibly bind small molecules such as oxygen or nitric oxide; they possess size and shape selectivity, the possibility of metalloenzyme mimicry, and immunomodulatory activity. These properties make them interesting for pharmaceutical industry and medicine. METHODS: The experiments were performed on mice. Different biochemical and molecular methods were used. RESULTS: Micronized zeolite (MZ) administered by gastric intubation to mice injected with melanoma cells significantly reduced the number of melanoma metastases. In mice fed MZ for 28 days, concentration of lipid-bound sialic acid (LSA) in serum increased, but lipid peroxidation in liver decreased. The lymphocytes from lymph nodes of these mice provoked a significantly higher alogeneic graft-versus-host (GVH) reaction than cells of control mice. After i.p. application of MZ, the number of peritoneal macrophages, as well as their production of superoxide anion, increased. However, NO generation was totally abolished. At the same time, translocation of p65 (NFkappaB subunit) to the nucleus of splenic cells was observed. CONCLUSION: Here we report antimetastatic and immunostimulatory effect of MZ and we propose a possible mechanism of its action. PMID- 11862471 TI - Establishment and characterization of a cell line from a malignant fibrous histiocytoma of bone developing in a patient with multiple fibrous dysplasia. AB - PURPOSE AND METHODS: In order to provide material for genetic analysis of fibrous dysplasia (FD), a cell line designated GBS-1 was established from a secondary bone malignant fibrous histiocytoma (MFH) developing in a rib of a 44-year-old male polyostotic FD patient. RESULTS: The GBS-1 cells are characterized by a pleomorphic spindle cell morphology with abundant mucus production. On transplantation to nude mouse subcutis the cell line forms myxoid-spindle cell sarcomas with giant cells, the myxoid product being positive for periodic acid Schiff (PAS) and alcian blue (Al-B) stains and completely digested by hyaluronidase, mimicking the original tumor. Chromosome and genetic analyses revealed multiple structural and numerical abnormalities of chromosomes with a large number of unidentifiable chromosomes and p53 mutation in exon 7 with LOH in the counterpart. CONCLUSIONS: Since cell lines for FD have hitherto not been available, the GBS-1 cells should prove useful for genetic analyses of FD and also MFH of bone origin. PMID- 11862472 TI - Granulomatous slack skin or granulomatous mycosis fungoides -- a case report. Complete response to percutaneous radiation and interferon alpha. AB - A 46-year-old man presented with circumscribed inflammatory poikilodermatic lesions of loose skin on the upper arm. Histologic examination disclosed a heavy lymphocytic infiltrate of the whole dermis and the upper part of the subcutaneous fat tissue with a predominant T-helper phenotype and about 10% of Mac 387 positive macrophages and some scattered multinucleated giant cells. Lymphocytic cells were aligned along the epidermal basement membrane and showed focal epidermotropism. In part these cells had multilobular nuclei. The diagnosis of granulomatous mycosis fungoides versus early granulomatous slack skin was made. The patient was treated with a combination of radiotherapy (total dose 36 Gy) and interferon-alpha as a maintenance treatment which resulted in complete remission and disease-free survival of 27 months up to now. PMID- 11862473 TI - Progesterone receptor status of breast cancer metastases. AB - PURPOSE: The usefulness of steroid receptor content in breast cancer metastases for metastatic disease therapy planning was examined in this study. METHODS: Steroid receptors in primary tumors and corresponding metastases in the same breast cancer patients ( n=23) were determined by five-point DCC assay. We carried out an analysis of the therapeutic response and comparison of the progression-free interval of patients treated with endocrine/chemo-endocrine therapy for metastatic disease according to the positive/negative progesterone receptor status of primary tumors, or of breast cancer metastases. RESULTS AND CONCLUSIONS: It seems that the lack of positive progesterone receptors in metastasis (0/8) and conversion from PR+ primary to PR- metastasis (5/8) may be important in describing the non-responder phenotype. We obtained a similar progression-free interval in patients with progesterone receptor positive/negative primary tumors, but a longer progression-free interval in the patients with progesterone receptor-positive metastases ( n=9) than with negative ones ( n=14), indicating the possibility of using steroid receptor content from metastases for metastatic disease therapy planning. PMID- 11862474 TI - Isolated brain metastases as the sole manifestation of a late relapse in breast cancer. AB - We report on a 62-year-old female patient suffering from breast cancer (invasive ductal, premenopausal, estrogen- and progesterone-receptor status unknown) first diagnosed in July 1991. After mastectomy and axillary lymphonodectomy (pT2 N2 (12/15) M0 G2), adjuvant chemotherapy consisting of six courses epirubicin and cyclophosphamide was performed. Since 1993, serum Ca 15-3 levels began to climb slowly. As of June 1996, Ca 15-3 was measured 50.1 U/ml (normal value <28.0 U/ml). In routine follow-ups, Ca 15-3 increased slowly, but no metastases of the breast cancer could be detected. In September 2000, the patient attended a routine follow-up examination with poor performance status, presenting with ataxia and a psychic syndrome. Computed tomography revealed multiple suprasellar brain metastases. We conclude that this is an unusual case of a very late clinical manifestation of sole brain metastases in a patient with breast cancer. A slow preceding increase of Ca 15-3-level over a period of 7 years was the only indicator of the upcoming brain metastases. PMID- 11862475 TI - Hyperthermia and mafosfamide in a human-derived malignant pleural mesothelioma cell line. AB - PURPOSE: Diffuse malignant pleural mesothelioma is the most common primary pleural malignancy. At the beginning of the last century, this tumor was of minor incidence. Meanwhile, the use of asbestos has led to and is still leading to a rise in pleural mesothelioma incidence. There is no standard therapy for this highly aggressive disease and the development of new therapeutic strategies is imperative. METHODS: We, therefore, investigated the morphological and pharmakokinetic effects of a combined thermochemotherapy consisting of the administration of different dosages of mafosfamide with and without the application of a 1-h hyperthermia at 41.7 degrees C on the human biphasic malignant pleural mesothelioma cell line MSTO-211H. After therapy, cells were prepared for light and electron microscopy. BrdU-incorporation for the S-phase fraction, TUNEL-labeling for detection of apoptosis, and quantitative assessments using the MTT assay were performed. RESULTS: Our results demonstrate that the combination of mafosfamide with hyperthermia leads to qualitatively and quantitatively enhanced cellular damage compared to monotherapy. During combined thermochemotherapy, cell damage and death is already induced at lower mafosfamide concentrations than without hyperthermia which suggests an additive effect from hyperthermia to the action of the alkylating drug mafosfamide. Cell death thereby mostly occurs as necrotic cell death rather than as apoptosis, although in a combined thermochemotherapy apoptosis is induced temperature-dependently, when comparing temperatures from 37 degrees C to 43 degrees C. CONCLUSIONS: We suggest that the effect of substances such as ifosfamide and cyclophosfamide which are in clinical use, might be enhanced by the combination of local or regional hyperthermia in order to improve the therapeutical index of these substances in the treatment of pleural mesothelioma. PMID- 11862476 TI - Expression of TNF-related apoptosis-inducing ligand (TRAIL) and its receptors in gastric carcinoma and tumor-infiltrating lymphocytes: a possible mechanism of immune evasion of the tumor. AB - PURPOSE: TNF-related apoptosis-inducing ligand (TRAIL) and its receptors have recently been known to be responsible for apoptotic signaling molecules in tumor cell lines and tissues. These molecules have been reported to be expressed on merely a transcription level, but not on a protein level. Moreover, little is known about TRAIL-mediated apoptosis in human carcinoma in vivo. METHODS: We investigated the presence and functional status of TRAIL and its receptors, DR4, DR5, and DcR2 on tumor as well as tumor-infiltrating lymphocytes (TIL) in primary ( n=37), and metastatic gastric carcinoma from malignant ascites ( n=37) by a flow cytometry. In addition, phenotypic proportions of major T-cell subsets or B cells in TIL were also determined. RESULTS: Membrane-bound TRAIL/its receptors are constitutively expressed at high levels in primary and metastatic carcinomas in nearly all the patients. Apoptotic tumor cells detected by terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick and labeling (TUNEL) were barely identified in primary and metastatic carcinomas. TIL in primary carcinoma showed a very low level of expression of TRAIL/its receptors and TUNEL-positive cells. In metastatic carcinoma, however, there was significant overexpression of TRAIL/its receptors in TIL associated with a higher frequency of apoptotic cell death detected by TUNEL. The TIL within metastatic carcinoma, but not within primary carcinoma, revealed the increased proportions of CD3(+) T cells bearing CD8(+)CD11b(-), CD8(+)CD11b(+), and CD4(+)CD62L(-), CD4(+)CD62L(+) surface phenotype in patients. CONCLUSIONS: These results suggest that TRAIL(+) and DcR2(+) metastatic carcinoma from malignant ascites could not only have resistance to DR4/DR5-induced apoptosis, but also might take the TRAIL-mediated counterattack against activated CD3(+) T cells. These functions of the cancer cells would neutralize host immune responses at the effector phase, and accelerate further invasion and/or metastasis of carcinoma through the escape from immune attack. PMID- 11862477 TI - Phosphatidylcholine does not protect rats against 5-fluorouracil/folinic acid induced damage of the intestinal luminal mucosa. AB - PURPOSE: The study was performed to evaluate the ability of phosphatidylcholine (PPC) to modulate the toxicity of 5-fluorouracil/folinic acid (5-FU/FA) in rats. METHODS: Twenty-eight female Wistar rats were divided into four series, each consisting of 6--8 animals and administered 0.5 ml isotonic solution i.v. once a day. The series were as follows: 1) saline; 2) 20 mg 5-FU + 4 mg FA; 3) 30 mg 5 FU + 6 mg FA; 4) 2 weeks daily orally 50 mg PPC/kg body weight (b.w.), on day 12, 30 mg 5-FU + 6 mg FA. As an indicator of membrane luminal impairment, 72 h after an i.v. administration, peptidase activities were measured by using an in situ perfusion model. RESULTS: The i.v. application of 5-FU/FA causes a clear reduction of enzyme activities in comparison to the control group. CONCLUSIONS: In investigations with rats, we have shown that a low concentration of oral PPC (50 mg/kg b.w.) cannot protect the luminal mucosa membrane from being injured by the enterotoxic action of 5-FU/FA. PMID- 11862478 TI - Androgen receptor status in female breast cancer: RT-PCR and Western blot studies. AB - PURPOSE: Although the androgen receptor (AR) has been found to be expressed in female breast tumours, its role in breast cancer remains unclear. In addition to the oestrogen receptor (OR) and progesterone receptor (PR), AR functional status may be important in the control of female breast cancer growth. METHODS: In order to define AR in breast cancer, 67 primary breast tumours and 8 normal breast samples as control tissue were analysed for AR expression at the mRNA and protein levels using RT-PCR and Western blotting, respectively. The association of AR expression with tumour size and axillary lymph node status was investigated. Expression of AR mRNA was compared with that of OR and PR. RESULTS: AR expression was identified in 66% (44/67) and 51% (34/67) of the cancers studied by RT-PCR and Western blotting, respectively. The number of positive samples and the level of AR mRNA were significantly higher among the cancer samples than among normal samples. No expression of AR protein was detected in normal breast tissue. A significant correlation between AR gene expression and its protein level in nuclei of carcinoma samples was demonstrated. The expression level of both AR gene and AR protein in nuclei was found to be positively correlated with tumour invasiveness. Of the breast carcinoma specimens, 44.8% (30/67) were OR-, PR- and AR-positive, while 14.9% (10/67) were steroid receptor-negative. However, 18% (12/67) of the primary breast tumours negative for OR and PR were positive for AR. CONCLUSIONS: The high incidence of AR expression in female breast tumours suggests a potential role of AR in breast cancer, in addition to OR and PR. PMID- 11862480 TI - Circadian variation of dihydropyrimidine dehydrogenase mRNA expression in leukocytes and serum cortisol levels in patients with advanced gastrointestinal carcinomas compared to healthy controls. AB - PURPOSE: The activity of dihydropyrimidine dehydrogenase (DPD) - the rate limiting enzyme in fluorouracil (5-FU) catabolism - has been reported to vary according to the time of day. On the basis of this data, so-called chronomodulated chemotherapy regimens with variable-rate infusions of 5-FU have been investigated in the treatment of advanced colorectal cancer. Recent results suggest lower toxicity of 5-FU by chronomodulated application. However, the pattern of circadian DPD activity levels have been shown to vary considerably. METHODS: We, therefore, studied the circadian changes in mRNA expression of DPD in leukocytes of ten patients with advanced gastrointestinal carcinomas prior to chronomodulated 5-FU-based salvage therapy and in 5five healthy controls. Simultaneously, we measured serum cortisol levels (SCL) to evaluate the endogenous circadian hormone rhythm. RESULTS: SCL displayed a consistent circadian rhythm with the mean peak value of serum cortisol at 8 a.m. and the mean trough value at 11 p.m. both in patients and in controls. However, mean minimum-maximum serum cortisol differences of SCL were significantly lower in patients compared to controls. In the 5fivehealthy controls, a trend towards a circadian rhythm of DPD mRNA expression was observed with the peak of expression at 5 a.m. which was significantly different from the trough at 2 p.m. ( P<0.005 Mann-Whitney-Wilcoxon test). When each control was studied separately, only two individuals showed circadian variations that could be fitted to a cosine wave ( P=0.001, P=0.014, Cosinor analysis). In contrast, DPD mRNA expression in patients with advanced gastrointestinal carcinomas did not demonstrate any consistent circadian rhythm. Pairwise comparisons of groups of DPD mRNA levels at different times of the day did not show significant differences. CONCLUSIONS: In conclusion, our analysis of DPD mRNA expression in leukocytes from healthy controls demonstrates first evidence for a circadian DPD mRNA expression periodicity. In patients with advanced gastrointestinal carcinomas, however, this rhythm seems to be disturbed although circadian endogenous cortisol secretion pattern is maintained. PMID- 11862479 TI - Glutathione S-transferase-pi in malignant tissues and plasma of human colorectal and gastric cancers. AB - PURPOSE: In this study, 16 paired samples of colorectal and gastric cancers and adjacent non-neoplastic tissues were analysed for the determination of glutathione S-transferase (GST) activities and the expression of GST-pi. METHODS: Western blotting procedure as well as plasma GST-pi levels were used. RESULTS: GST activities were found to be increased in malignant tissues of patient compared with adjacent normal tissues. A significant correlation was detected between GST activity and GST-pi expression in malignant tissues of patients. Plasma GST-pi levels increased in patients compared to aged-matched control subjects. When the patients were grouped according to TNM stage, GST-pi expression in malignant tissues as well as plasma GST-pi levels were higher in patients with more advanced tumor stages. CONCLUSIONS: Our results indicate that GST-pi expression in malignant tissues and plasma GST-pi levels in human colorectal and gastric cancers increased depending on the stages of tumor. PMID- 11862481 TI - Galectin fingerprinting by immuno- and lectin histochemistry in cutaneous lymphoma. AB - Owing to their relevance for growth regulation and cell adhesion monitoring the expression of galectins (endogenous beta-galactoside-binding lectins) and their binding sites has relevance for tumor biology. Using galectin-type-specific reagents (non-crossreactive antibodies for proto-type galectin-1, chimera-type galectin-3 and tandem-repeat-type galectins-4 and -8, and labeled galectins-1, 3, and -4) we determined galectin expression in cutaneous T cell lymphomas (CTCL) and controls. In addition to commonly studied galectins-1 and -3, tandem-repeat type galectins could be detected, i.e., galectin-8 in six from 15 cases and galectin-4 in one of 15 cases. In view of relevant ligands such as bcl-2 or integrins the presence of galectins-3 and -8 seems to be possibly related to loss of proliferation control and change in cell adhesion properties that are involved in clonal expansion and epidermal spread of malignant T cell clones. Successful chemotherapy of CTCL alters galectin expression selectively as shown for liposomal doxorubicin. PMID- 11862482 TI - Regression of brain metastases from breast carcinoma after chemotherapy with bendamustine. AB - PURPOSE: Bendamustinehydrochloride (bendamustine) is an alkylator with anticipated antimetabolic activity. It has shown activity in malignant lymphoma and breast cancer. Up to now there are no reports about the activity of bendamustine in the treatment of brain metastases. We report on a 38-year-old woman with brain metastases from breast cancer. The first diagnosis of an invasive ductal, hormone receptor negative breast cancer was made in October 1997, stage pT1c pN2 (11/11)cM0 - G3. After lumpectomy and resection of axillary lymph nodes, the patient received adjuvant chemo- and radiotherapy. Twenty six months after the first diagnosis, bone metastases occurred and were treated with radiotherapy. One month later, multiple liver metastases developed which were treated with trastuzumab and paclitaxel. Four months later, progress of the liver metastases and malignant infiltration of bone marrow with thrombopenia occurred. METHODS: Chemotherapy with bendamustine at a dose of 150 mg/m(2) on day 1 and 2 was initiated. Two days later, the patient suffered from central facial palsy and subsequent computed tomography (CT) revealed three brain metastases in the frontal, parietal and occipital region. Because of the advanced liver metastases with clinical and laboratory signs of liver insufficiency and the reduced performance status of the patient, chemotherapy with bendamustine was continued and no local treatment of the brain metastases was performed. RESULTS: After two courses of bendamustine, ultrasound showed regression of the liver metastases. Liver enzymes decreased, platelets increased, and the patient's performance status improved. Additionally, two of the three brain metastases were no longer detectable by CT, the third had decreased compared to the time of diagnosis. CONCLUSION: To the best of our knowledge, this is the first report describing major activity of bendamustine in cerebral metastases. Thus, it may be considered as another therapeutic strategy against metastatic brain cancer. However, this finding warrants further investigation in clinical trials. PMID- 11862483 TI - Markers of drug resistance in relapsing colon cancer. AB - PURPOSE: 5-Fluorouracil failure and drug resistance, which often occurs during chemotherapy, is still a great obstacle to the success of human colon cancer treatment. Thus, the comparative study of markers of drug resistance in cancer cells before and after chemotherapy may be extremely helpful in the selection of the appropriate chemotherapeutic drug in colon cancer patients who fail adjuvant treatment with 5-fluorouracil. In the present study we examined the differential expression of three multidrug resistance-related proteins (i.e., P-glycoprotein, MRP, and LRP) and of topoisomerase IIalpha in a series of 20 primary colon carcinomas and their recurrences. METHODS: All markers were determined at tissue level by three-step immunohistochemistry using appropriate monoclonal antibodies, and the markers' immunopositivity was quantified by image analysis. In addition, Feulgen stain was used for the assessment of nuclear DNA content of malignant cells at their primary location. RESULTS: Some degree of aneuploidy was detected in all primary carcinomas. The immunoexpression of the three multidrug resistance related proteins did not change significantly, either qualitatively (positivity vs negativity) or quantitatively, after chemotherapy. On the contrary, the percentages of topoisomerase IIalpha-positive malignant cells were significantly increased in the tumour recurrences by comparison to their primary locations ( P=0.011). CONCLUSIONS: According to our results, increased topoisomerase IIalpha immunohistochemical expression appears to be part of the malignant cells' phenotype in recurrent colon cancers. Therapeutic options after failure of 5 fluorouracil-based treatment could therefore include appropriate topoisomerase IIalpha-targeted drugs. PMID- 11862484 TI - UbcD4, a ubiquitin-conjugating enzyme in Drosophila melanogaster expressed in pole cells. AB - The ability to destroy a particular protein at a particular time is central to the regulation of many cellular processes. Selective proteolysis in eukaryotic cells is carried out primarily by the ubiquitin-proteasome pathway. Attachment of a ubiquitin polymer to an unwanted protein causes it to be degraded by the proteasome. Several classes of enzyme, known as E1s, E2s and E3s, control the stepwise formation of a ubiquitin-protein conjugate. The specificity of substrate selection lies with the E2s and E3s. Here we describe the cloning of a Drosophila E2 gene, UbcD4, which is only expressed in embryos. Its expression pattern in stage 10-11 embryos suggests a role in germ cell development. UbcD4 can interact with the polyubiquitin-binding subunit of the proteasome. PMID- 11862485 TI - Mutants of the carotene cyclase domain of al-2 from Neurospora crassa. AB - Phytoene synthase and carotene cyclase, two key enzymes in carotenoid biosynthesis, are encoded by two separate genes in bacteria and plants, but by a single bifunctional gene in fungi. The cyclase function has been demonstrated for the products of the genes crtYB from the basidiomycete Xanthophyllomyces dendrorhous, and for carRA and carRP from the zygomycetes Phycomyces blakesleeanus and Mucor circinelloides, respectively. These three genes are highly similar to al-2 from Neurospora crassa. Taking advantage of the high proportion of the final product of the carotenoid pathway that accumulates Neurospora when mycelium is illuminated at low temperature, we have isolated two mutants with a pale reddish pigmentation. Both mutants are complemented by the wild-type al-2 gene, and carry mutations in the al-2 domain to which cyclase activity has been attributed in other fungi. The mutants lack neurosporaxanthin and accumulate an unidentified reddish carotenoid, as shown by column chromatography and HPLC. The chemical and spectrophotometrical properties of this carotenoid are consistent with the absence of carotenoid cyclization, and indicate that the product of al-2 is bifunctional. The existence of a single gene responsible for phytoene synthase and carotene cyclase thus seems to be a widespread trait among filamentous fungi, as shown by the examples now known in a basidiomycete, two zygomycetes and one ascomycete. PMID- 11862486 TI - Suppression of heterochromatic gene variegation can be used to distinguish and characterize E(var) genes potentially important for chromosome structure in Drosophila melanogaster. AB - Hundreds of genic modifiers of position effect variegation (PEV) have been isolated in Drosophila melanogaster with a view to identifying genes important for chromosome structure. Here we propose a supplementary genetic screen to pinpoint candidate genes that are most likely to function in chromosome organization, within the enhancer of variegation [E(var)] class of modifiers. Our strategy takes advantage of the fact that variegating euchromatic and heterochromatic genes respond oppositely to changes in the dosage of heterochromatin proteins. Consequently, only when enhancement of euchromatic gene variegation results from increased formation of heterochromatin should suppression of heterochromatic gene variegation be observed. Mutations in four E(var) genes were tested for the ability to suppress variegation of multiple alleles of the heterochromatic light ( lt) gene in a variety of tissues and at several developmental stages. Mutations in E(var)3-4, E(var)3-5 and modifier of mdg4 [ mod(mdg4)] suppressed lt variegation. In contrast, a mutation in the Trithorax-like ( Trl) gene, which encodes GAGA factor, enhanced or had no effect on lt variegation, consistent with its known role in promoting transcription. These data show that suppression of lt variegation can be used as an assay to distinguish between members of the E(var) class of modifiers. PMID- 11862487 TI - Genetic control of fruit shape acts prior to anthesis in melon ( Cucumis melo L.). AB - Genetic control of fruit shape in Cucumis melo was studied using QTL analysis in two Recombinant Inbred (RI) populations consisting of 163 and 63 individuals, respectively, obtained by crossing the same round-fruited parent with two different elongated-fruit lines. Fruit shape is mainly explained by fruit length in these two populations. Most QTLs for fruit shape and ovary shape detected were found to co-segregate, thus demonstrating early control of fruit shape during ovary development. A high level of correlation between fruit shape and ovary shape was also found in 14 unrelated genetic lines, a finding which suggests that control of fruit shape by gene(s) active early in the ovary is a general feature in C. melo. Two major flower genes, a ( monoecious) and p ( pentamerous), were shown to have major effects on fruit shape. Major tightly linked QTLs for fruit and ovary shape were found close to the a and p genes, probably reflecting their pleiotropic effect on fruit shape. Moreover, one of the two QTLs detected in the Vedrantais x PI 414723 population was also found in the Vedrantais x PI 161375 population. Variation of fruit shape in melon could be due to variations having quantitative effects on a large set of genes that are probably involved in ovary development. PMID- 11862488 TI - A novel MADS-box gene subfamily with a sister-group relationship to class B floral homeotic genes. AB - Class B floral homeotic genes specify the identity of petals and stamens during the development of angiosperm flowers. Recently, putative orthologs of these genes have been identified in different gymnosperms. Together, these genes constitute a clade, termed B genes. Here we report that diverse seed plants also contain members of a hitherto unknown sister clade of the B genes, termed B(sister) (B(s)) genes. We have isolated members of the B(s) clade from the gymnosperm Gnetum gnemon, the monocotyledonous angiosperm Zea mays and the eudicots Arabidopsis thaliana and Antirrhinum majus. In addition, MADS-box genes from the basal angiosperm Asarum europaeum and the eudicot Petunia hybrida were identified as B(s) genes. Comprehensive expression studies revealed that B(s) genes are mainly transcribed in female reproductive organs (ovules and carpel walls). This is in clear contrast to the B genes, which are predominantly expressed in male reproductive organs (and in angiosperm petals). Our data suggest that the B(s) genes played an important role during the evolution of the reproductive structures in seed plants. The establishment of distinct B and B(s) gene lineages after duplication of an ancestral gene may have accompanied the evolution of male microsporophylls and female megasporophylls 400-300 million years ago. During flower evolution, expression of B(s) genes diversified, but the focus of expression remained in female reproductive organs. Our findings imply that a clade of highly conserved close relatives of class B floral homeotic genes has been completely overlooked until recently and awaits further evaluation of its developmental and evolutionary importance. Electronic supplementary material to this paper can be obtained by using the Springer Link server located at http://dx.doi.org/10.1007/s00438-001-0615-8. PMID- 11862489 TI - Isolation and characterization of repetitive DNA sequences from Panax ginseng. AB - Repetitive sequences constitute a significant component of most eukaryotic genomes, and the isolation and characterization of repetitive DNA sequences provide an insight into the organization and evolution of the genome of interest. We report the isolation and characterization of the major classes of repetitive sequences from the genome of Panax ginseng. The isolation of repetitive DNA from P. ginseng was achieved by the reannealing of chemically hydrolyzed (200 bp-1 kb fragments) and heat-denatured genomic DNA to low C(o)t value. The low C(o)t fraction was cloned, and fifty-five P. ginseng clones were identified that contained repetitive sequences. Sequence analysis revealed that the fraction includes repetitive telomeric sequences, species-specific satellite sequences, chloroplast DNA fragments and sequences that are homologous to retrotransposons. Two of the retrotransposon-like sequences are homologous to Ty1/ copia-type retroelements of Zea mays, and six cloned sequences are homologous to various regions of the del retrotransposon of Lilium henryi. The del retrotransposon-like sequences and several novel repetitive DNA sequences from P. ginseng were used to differentiate P. ginseng from P. quinquefolius, and should be useful for evolutionary studies of these disjunct species. PMID- 11862490 TI - Recombinational error and deletion formation in Neisseria gonorrhoeae: a role for RecJ in the production of pilE (L) deletions. AB - Genetic linkage within Neisseria gonorrhoeae populations is in equilibrium, yet the physical linkage map indicates a relatively stable chromosome structure, despite an apparently vast potential for mispairing between repeated sequences (e.g. between the multiple pil or opa alleles, or through mispairing of any of the numerous small repeated sequences that are liberally scattered throughout the chromosome). Therefore, the stability of the physical linkage map suggests that aberrant recombination between repeated sequences is a rare event. This study was undertaken to explore some of the parameters that may govern deletion events between short direct oligonucleotide repeats, using a chromosomal locus that appears to be especially prone to deletions (the pilin expression locus; pilE). In this report, we demonstrate that deletion formation at pilE occurs primarily through recombinational error following a pilE/pilS interaction; illegitimate (i.e. RecA-independent) events can occur, but they are infrequent. In contrast, when genetically engineered opa deletion substrates were constructed and placed in the chromosome, deletions at the opa loci were infrequent even under rec(+) conditions. A model is presented in which the gonococcal RecA and RecJ proteins promote pilE deletions through a recombination event that is templated or stabilised by a pilE/pilS interaction. PMID- 11862491 TI - mRNA stability and the secretion signal of HrpA, a pilin secreted by the type III system in Pseudomonas syringae. AB - Gram-negative bacteria that are pathogenic for animals or plants utilise a specialised Type III secretion system to inject effector proteins into their eukaryotic target cells. The basis for selection of the proteins to be translocated via type III systems is still enigmatic. No clearly defined consensus amino acid sequence that could serve as a specific secretion signal has been identified, and the hypothesis that an mRNA secondary structure acts as the signal has several shortcomings. We have localised a secretion signal that is sufficient to ensure the secretion of the pilin HrpA, a substrate and an indispensable extracellular component of the type III secretion machinery of Pseudomonas syringae pv. tomato DC3000, to the first 15 codons. Transcription of hrpA starts at a single site 42 bp upstream of the first codon. Gene swapping experiments revealed that altering the continuity of the 5' non-translated leader with the region including the secretion signal radically decreased accumulation of the hrpA transcript. These results indicate that an mRNA secondary structure, possibly formed in this region, is important for efficient expression of the gene. The proposed secondary structure is not, however, indispensable for the secretion of HrpA and it does not couple secretion and translation. PMID- 11862492 TI - A bacterial artificial chromosome (BAC) library for sunflower, and identification of clones containing genes for putative transmembrane receptors. AB - Sunflower (Helianthus annuus L.) is an economically important oil seed crop with an estimated genome size of 3000 Mb. We have constructed a bacterial artificial chromosome (BAC) library for sunflower, which represents an estimated 4- to 5 fold coverage of the genome. Nuclei isolated from young leaves were used as a source of high-molecular-weight DNA and a partial restriction endonuclease digestion protocol was used to cleave the DNA. A random sample of 60 clones indicated an average insert size of 80 kb, implying a 95% probability of recovering any specific sequence of interest. The library was screened with chloroplast DNA probes. Only 0.1% of the clones were identified to be of chloroplast origin, indicating that contamination with organellar DNAs is very low. The utility of the library was evaluated by screening for the presence of genes for putative transmembrane receptors sharing epidermal growth factor (EGF) and integrin-like domains. First, a homologous sunflower EST (HaELP1) was obtained by degenerate RT-PCR cloning, using Arabidopsis thaliana genes (AtELP) as a source of consensus sequences. Three different BACs yielded positive hybridization signals when HaELP1 was used as a probe. BAC subcloning and sequencing demonstrated the presence of two different loci putatively homologous to genes for transmembrane proteins with EGF- and integrin-like domains from sunflower. This work demonstrates the suitability of the library for homology map based cloning of sunflower genes and physical mapping of the sunflower genome. PMID- 11862493 TI - A dominant mitochondrial mutator phenotype of Saccharomyces cerevisiae conferred by msh1 alleles altered in the sequence encoding the ATP-binding domain. AB - In order to improve our understanding of the role of the yeast MSH1 gene in error avoidance in mitochondrial DNA, two msh1 alleles were constructed, which encode proteins with amino acid substitutions in an ATP-binding domain that is highly conserved among MutS homologs. Here, we report that moderate overexpression of the msh1-R813W or msh1-G776D allele, in strains which also carry the wild-type MSH1 allele, slightly increases the frequency of mutations conferring resistance to erythromycin (E(r)) and elevates the frequency of alterations within a polyGT tract present in mitochondrial DNA (mtDNA). This result indicates that the mutant alleles confer a dominant mitochondrial mutator phenotype and strongly suggests that the ATP-binding domain plays a crucial role in the in vivo function of Msh1p. Interestingly, we have found that overexpression of wild-type MSH1 has opposite effects on the stability of polyGT vs. polyAT tracts present in mtDNA; excess of Msh1p slightly increases the stability of polyGT tracts, whereas the stability of polyAT tracts is dramatically decreased. We show that although overexpression of msh1-R813W or msh1-G776D also results in a marked overall increase in the frequency of alterations in polyAT tracts, the spectrum of alterations differs from that found in cells overexpressing MSH1; large deletions predominate in the latter case, while 2-bp deletions are generated in cells that overproduce the mutant msh1p. This result strongly suggests that the mutations in the ATP binding domain change the specificity of the protein with respect to the recognition of potentially mutagenic structures in mtDNA. PMID- 11862494 TI - AtE2F-a and AtDP-a, members of the E2F family of transcription factors, induce Arabidopsis leaf cells to re-enter S phase. AB - In eukaryotes, transcription factors of the E2F family, in addition to having a role in cell proliferation, participate in regulating apoptosis, differentiation and development. In Arabidopsis thaliana, eight gene sequences have been identified as encoding E2F or DP homologues. DP proteins form heterodimers with E2Fs. The aim of this work was to characterize the functions of three of these factors: AtE2F-a, AtE2F-b and AtDP-a. Here we report that AtE2F-a and AtE2F-b transactivate a reporter gene via an E2F consensus cis-acting element in Arabidopsis protoplasts. AtE2F-a is a more potent activator than AtE2F-b. Furthermore, co-expression of the E2F partner AtDP-a, or the DNA binding protein AtPur alpha, modulates the activation of AtE2F-a. Taken together, these results suggest that AtE2F-a, AtE2F-b and AtDP-a share features characteristic of members of the E2F family of transcription factors. Moreover, over-expression of AtE2F-a and AtDP-a can induce differentiated, non-dividing, leaf cells to re-enter S phase. We conclude that the transcription factor AtE2F-a plays an important role in progression into S phase, which probably correlates with its capacity to stimulate transcription. PMID- 11862495 TI - A novel cis-acting cysteine-responsive regulatory element of the gene for the high-affinity glutathione transporter of Saccharomyces cerevisiae. AB - We cloned a DNA fragment from Saccharomyces cerevisiae that complemented the deficiency in high-affinity glutathione transport activity conferred by a gsh11 mutation, and found that the ORF responsible was YJL212c, which had already been designated as OPT1 and HGT1 by others. Northern analysis clearly demonstrated that this ORF, now referred to as OPT1/ HGT1/ GSH11, was induced by sulfur starvation and repressed by adding cysteine to the growth medium. Reporter gene assays showed that a segment spanning the region between positions -371 and -355 was essential for the regulation of this gene. A sequence of 9 nt, CCGCCACAC (from -364 to -356), in this region was shown to be required for protein binding, using an electrophoretic mobility shift assay. Based on these results, we propose that CCGCCACAC comprises the core of a cis-acting element involved in cysteine responsive gene regulation in S. cerevisiae. PMID- 11862496 TI - Genetic mapping of the non-nodulation phenotype of the mutant MN-1008 in tetraploid alfalfa (Medicago sativa). AB - Abstract. Roots of the non-nodulating Medicago sativa mutant MN-1008 neither undergo root-hair curling, cortical cell division nor any of the early molecular events that accompany nodule initiation and development following rhizobial infection or treatment with Nod factor. These observations suggested that the mutation(s) impaired a pivotal function in Nod factor perception or in the signal transduction pathway. In this paper we show that the genetic lesion conditioning the recessive non-nodulation phenotype in the tetraploid alfalfa mutant MN-1008 can be localized to a single region on LG5 of the M. sativa genetic map. This conclusion is based on genetic analyses conducted at the tetraploid level, involving both segregation analysis and genetic mapping of the trait with respect to molecular DNA markers. The genetic mapping of the Nod(-) phenotype was performed in a segregating tetraploid F2 population, taking advantage of the availability of an advanced genetic map for diploid alfalfa. Two tightly linked flanking markers have been identified which will facilitate the physical mapping and cloning of the gene(s) that underlie(s) the non-nodulation phenotype. PMID- 11862497 TI - Nausea and emesis: still an unsolved problem in cancer patients? PMID- 11862498 TI - Delayed emesis: incidence, pattern, prognostic factors and optimal treatment. AB - Delayed emesis has been arbitrarily defined as vomiting and/or nausea beginning, or persisting for, more than 24 h after chemotherapy administration. Acute emesis is the most important prognostic factor for delayed emesis. Owing to the relatively high incidence and severity all patients treated with cisplatin > or = 50 mg/m(2) should receive antiemetic prophylaxis. In these patients a combination of dexamethasone plus metoclopramide or a 5-HT3 antagonist is the most efficacious regimen. All patients submitted to moderately emetogenic chemotherapy, such as cyclophosphamide, carboplatin, doxorubicin and epirubicin, should also receive antiemetic prophylaxis with oral dexamethasone to prevent delayed emesis. PMID- 11862499 TI - Nausea and emesis: evidence for a biobehavioral perspective. AB - Abstract. For most people, nausea and vomiting (NV) are simply unfortunate consequences of overindulgent college days or overenthusiastic amusement rides. Yet for most cancer patients, nausea and vomiting (also referred to as emesis) remain among the most frequent side effects of cancer chemotherapy. Patients typically view control of nausea as more important than control of emesis, while physicians and nurses judge emesis control to be more important to antiemetic efficacy than nausea control. The 5-HT3 receptor antagonists have been shown to be clinically more effective in controlling emesis, particularly that caused by regimens containing high-dose cisplatin, than previously available agents. Disappointingly, however, these drugs do not appear to be more effective than previous antiemetics in reducing nausea. In addition, the 5-HT3 receptor antagonists may become less effective over repeated chemotherapy administrations, and they remain expensive. An impediment to research progress has been an insularity that has prompted two parallel research efforts: one searching for biological understanding to enhance pharmacological intervention(s) and the other searching for psychological understanding to aid in developing more effective behavioral intervention(s). While both approaches have been successful, it is time to have the two views merge into a biobehavioral framework that combines them both. This paper draws on both physiological and psychological origins of NV to begin the development of a biobehavioral model of development that integrates features of both approaches. PMID- 11862500 TI - How do we manage patients with refractory or breakthrough emesis? AB - There is evidence that, in spite of the Perugia consensus, acute and delayed emesis are treated in a suboptimal way. Thus breakthrough and refractory emesis as defined in this paper may be related to inadequate therapy. Several interventions have been used in attempts to stop breakthrough emesis, including use or repeat use of setrons, corticosteroids, D2-receptor antagonists including neuroleptics, or sedatives. It has been documented that refractory emesis responds to various modifications of the original antiemetic regimen, including the addition of a D2-receptor antagonist or a switch to another setron. In conclusion, no level I or II evidence-based guidelines can be given, as few adequate studies have been performed in this area, which therefore remains poorly documented. PMID- 11862501 TI - Is there a role for melatonin in supportive care? AB - Melatonin (MLT) is the main hormone released from the pineal gland and has proved to have physiological antitumor activity. MLT has been shown to exert anticancer activity through several biological mechanisms: antiproliferative action, stimulation of anticancer immunity, modulation of oncogene expression, and anti inflammatory, anti-oxidant and anti-angiogenic effects. Several experimental studies have shown that MLT may inhibit cancer cell growth, and preliminary clinical studies seem to confirm its anticancer property in humans. In addition, MLT may have other biological effects, which could be useful in the palliative therapy of cancer, namely anticachectic, anti-asthenic and thrombopoietic activities. On this basis, the present clinical investigation was performed in an attempt at better definition of the therapeutic properties of MLT in human neoplasms. In a first clinical study, we evaluated the effects of MLT in a group of 1,440 patients with untreatable advanced solid tumors, who received supportive care alone or supportive care plus MLT. In a second study, we evaluated the influence of MLT on the efficacy and toxicity of chemotherapy in a group of 200 metastatic patients with chemotherapy-resistant tumor histotype, who were randomized to receive chemotherapy alone or chemotherapy plus MLT. In both studies, MLT was given orally at 20 mg/day during the dark period of the day. The frequency of cachexia, asthenia, thrombocytopenia and lymphocytopenia was significantly lower in patients treated with MLT than in those who received supportive care alone. Moreover, the percentage of patients with disease stabilization and the percentage 1-year survival were both significantly higher in patients concomitantly treated with MLT than in those treated with supportive care alone. The objective tumor response rate was significantly higher in patients treated with chemotherapy plus MLT than in those treated with chemotherapy alone. Moreover, MLT induced a significant decline in the frequency of chemotherapy-induced asthenia, thrombocytopenia, stomatitis, cardiotoxicity and neurotoxicity. These clinical results demonstrate that the pineal hormone MLT may be successfully administered in medical oncology in the supportive care of untreatable advanced cancer patients and for the prevention of chemotherapy induced toxicity. PMID- 11862502 TI - Symptom control in cancer patients: the clinical pharmacology and therapeutic role of suppositories and rectal suspensions. AB - Rectally administered medications are essential in palliative medicine, particularly in the last days of life. They are underutilized. The pharmacology of rectally administered medications relates not only to the medication but also to the suppository base, additives, drug ionization, p K(a), absorptive surface of the rectum, and rectal health. The pharmacokinetics may differ from those of orally administered medications owing to reduced hepatic first-pass clearance. In this review the pharmacology of rectally administered palliative medications is reviewed and the use of individual drugs is outlined. PMID- 11862504 TI - 'Enough is enough': qualitative findings on the impact of dexamethasone during reinduction/consolidation for paediatric acute lymphoblastic leukaemia. AB - The results of a longitudinal study conducted with children undergoing treatment for acute lymphoblastic leukaemia (ALL) and their families in Brisbane, Australia, indicate that the emotional impact of one of the protocol drugs, dexamethasone, is acutely distressing. The findings presented cover the second interviews with the parents and children of the first 11 ALL families to have completed the re-consolidation stage of treatment. The results indicate that the negative impact of this drug is particularly severe during the reconsolidation stage, when families are exhausted with coping with the intensity of treatment. Thus, the administration of dexamethasone is a critical point in the pathway of care for children with ALL. The emotional consequences of the drug are profoundly disturbing, not only for the child, but for the whole family. The findings indicate that the period when dexamethasone is being administered is an important time for providing families with emotional support and information about likely sequelae of treatment. Because of guilt and self-doubt parents will not necessarily seek help, even if it is greatly needed. Recommendations are provided as to possible ways of reducing the distressing impact of the administration of this pharmaceutical intervention. PMID- 11862503 TI - Pretreatment factors predicting the development of postchemotherapy nausea and vomiting in Chinese breast cancer patients. AB - A prospective longitudinal study was designed to assess the role of pretreatment proneness to nausea and vomiting (NV) in the development of postchemotherapy NV in a group of Chinese breast cancer patients receiving moderately highly emetogenic chemotherapy. Seventy-one chemotherapy-naive subjects participated in the study. Patients were assessed the day before chemotherapy with measurements of their anxiety level, depression, fatigue and proneness to NV, motion sickness, NV experienced in past pregnancies, history of labyrinthitis, expectation of developing NV and expectation of developing pain. Patients also completed daily assessments of frequency, duration and intensity of NV for the 7 days after chemotherapy. Regression analyses revealed that nonpharmacological factors explained part of the variance of NV, the most common predictors being a history of labyrinthitis, expectation of developing NV after chemotherapy, younger age, stage of disease, and state anxiety. The explanatory power of the models ranged from 6% to 23% of the variance of the independent variable. There were different explanatory models for acute and delayed NV. Results indicate that consideration of the role of nonpharmacological factors in the development of NV could lead to more effective management of NV induced by chemotherapy. PMID- 11862505 TI - Double-flash, large-fraction radiation therapy as palliative treatment of malignant superior vena cava syndrome in the elderly. AB - Rapid control of symptoms is mandatory in cancer-induced superior vena cava syndrome (SVCS), but older patients often do not tolerate aggressive approaches. In order to maximize symptom relief and minimize treatment-related discomfort of aged patients in poor health we adopted a short-course, large-fraction radiation therapy (RT) schedule. Twenty-three consecutive patients aged over 70 who were suffering from solid-malignancy-related SVCS were enrolled. A total dose of 12 Gy was given in two 6-Gy fractions, 1 week apart, mainly in an out-patient setting. Completion of therapy to give up to 37-40 Gy was planned in the best-responding patients. Symptom relief was experienced by 8 patients as early as 4-5 days after the first fraction. The overall response rate was 87%. Despite some mild systemic side effects (chest pain, fever) reported by 5 patients (22%), overall toxicity was negligible. Short-course, double-flash RT stands as an effective and safe tool in the palliative treatment of malignant SVCS in older patients. Fractions larger than 6 Gy can be avoided in order to minimize side and toxic effects. PMID- 11862506 TI - Communicating prognosis to patients with metastatic disease: what do they really want to know? AB - Clinical audits suggest that prognosis is often not discussed with cancer patients, and cancer patients often over- or underestimate their prognosis. Doctors wish to convey information honestly and sensitively, but are often unsure how best to achieve this. An evidence base and guidelines in this area are lacking. This study aimed to obtain patient and health professional views on optimal ways of presenting prognosis to patients with metastatic breast cancer. Qualitative methods were used to generate participant views and experiences. Seventeen patients and 13 health professionals working in cancer care participated in structured interviews, which were audio-taped and transcribed. Sampling was discontinued when informational redundancy was achieved. The transcribed interviews were content analysed by a trained assessor using the constant-comparative method. Seven primary themes were identified, including: communication within a caring, trusting, long-term relationship; open and repeated negotiations for patient preferences for information; clear, straightforward presentation of prognosis where desired; strategies to ensure patient understanding; encouragement of hope and a sense of control; consistency of communication within the multi-disciplinary team; communication with other members of the family. Communication about prognosis in a metastatic setting requires considerable resources from both patients and doctors. Nevertheless, a number of useful strategies were identified. A quantitative study obtaining feedback from a large and representative sample of patients with metastatic cancer is now in progress, to confirm patient preferences for the communication styles identified in this qualitative study. PMID- 11862507 TI - Cancer patient education in Iran: a descriptive study. AB - Abstract. This study was carried out to examine the status of cancer patient education in Iran. Using the Multinational Association of Supportive Care in Cancer's (MASCC) patient education questionnaire, 310 individuals - a sample of heterogeneous cancer patients ( n=167) and their relatives ( n=143) - were enrolled in the study. The pooled results indicated that only 15% of respondents believed more than 80% of cancer patients were told of their diagnosis. In contrast, 30% of respondents thought less than 20% of patients knew their cancer diagnosis. When asked, "Were you given written materials about (i) cancer, (ii) treatment, and (iii) symptom management", the vast majority of respondents said "No" (91%, 87%, and 87%, respectively). When respondents were asked, "Would you like to learn more about cancer and treatments", 97% said "Yes". Most respondents indicated the need for information on the treatments available (27%) and general information about cancer (20%); most had sought information from health professionals (31%), other cancer patients and friends (29%), and television (22%). Finally, it was found that concern about patients' depression (17%), lack of printed materials (13%), the idea that it was better for patients not to know (12%), and families' requests not to tell the patient (11%) were the most frequently stated barriers to or reasons for restricted cancer patient education. The findings of the study suggest that cancer patient education in Iran is very poor and there is an urgent need to develop policy guidelines on disclosure of cancer diagnoses and patient education. PMID- 11862508 TI - Can a soft diet prevent bowel obstruction in advanced pancreatic cancer? AB - Although many patients are advised to follow a high-fiber diet to avoid constipation, it seems that a soft diet such as that recommended after bowel surgery may well be more helpful in avoidance of intestinal obstruction. PMID- 11862510 TI - Follow-up of extended pterional orbital decompression in severe Graves' ophthalmopathy. AB - BACKGROUND: Compressive optic neuropathy (CON) with visual loss is, apart from corneal exposure and disfigurating proptosis, the most serious clinical sign encountered in Graves' ophthalmopathy. However, numerous different approaches and operative techniques have been proposed for orbital decompression, with varying results and side effects. The purpose of the present study was to analyze peri operative data and long-term results in patients with severe thyroid-related orbitopathy, treated by extended pterional orbital decompression, comparing its effectiveness to other procedures. METHOD: An extended pterional orbital decompression was performed in 42 consecutive patients (59 orbits) with severe thyroid-associated ophthalmopathy after failure of medical and radiation therapy during an 11-year period. Pre- and postoperative examination included visual acuity, Hertel exophthalmometry, ocular motility, visual fields (Goldmann perimetry) and notification of complications. Long-term evaluation was carried out on average at 11 months postoperatively (range 5-26 months). FINDINGS: Visual acuity improved rapidly from a preoperative average of 0.53 +/- 0.33 (range, 0-1) to 0.77 +/- 0.31 (range, 0-1) postoperatively (p<0.001). Worsening was not seen. An average reduction of proptosis of 3.79 +/- 2.32 mm (range, 0.5-8 mm) was achieved with a mean preoperative Hertel measurement of 24.7 +/- 3.93 mm (range, 15-33 mm) (p<0.001). Double vision and restriction of eye motility was present in 76.3% of patients preoperatively and improved in 63% of patients (p<0.001). No new onsets of not already pre-existing double vision was seen. Complications included two cases of permanent palsy of the frontal branch of the facial nerve. INTERPRETATION: The extended pterional orbital decompression improved vision and decreased proptosis and restriction of extra-ocular movements in patients with severe sight-threatening and disfiguring cases of Graves' orbitopathy and is still an effective and low-risk alternative to other non-neurosurgical operative techniques. Especially new developing postdecompression strabismus can be successfully avoided. PMID- 11862511 TI - The effect of posterior instrumentation following PLIF with BAK cages is most pronounced in weak bone. AB - BACKGROUND: The use of BAK-cages for lumbar fusion has become very popular but complications such as cage subsidence and settling occurred. To treat these complications posterior instrumentation was used to improve segmental stability. It is, however, poorly understood, why some patients require additional posterior instrumentation, whereas the majority do not. The objectives of the study presented were first to determine the influence of bone mineral density (BMD) to the initial compressive stiffness of a segment that underwent posterior lumbar interbody fusion (PLIF) with two BAK-cages. Second, to estimate the importance of additional posterior instrumentation for compressive stiffness with respect to bone mineral density. METHODS: A validated finite element model (FEM) including posterior decompression and stabilisation by two BAK-cages (BAK_FEM) was used to predict the initial compression stiffness in axial loading of 600 N. This model was used to predict the influence of various grades of BMD on compression stiffness. A second FEM was generated in which additional posterior screw-rod instrumentation was simulated (BAK+PI_FEM) and this model used to predict the influence of BMD in axial loading. FINDINGS: The responses of all FEM suggested that initial compressive stiffness will increase if there is an increase of BMD. The stiffness as predicted by BAK+PI_FEM was always superior to FEM_BAK. This difference was most pronounced for weak bone quality. INTERPRETATION: Compression stiffness following PLIF with BAK-cages depends on BMD. Additional posterior instrumentation results in an additional increase of compression stiffness. This effect is most pronounced in simulated soft bone quality. These results may help to select patients for combined stabilisation. PMID- 11862512 TI - Effects of the temporary clipping in aneurysm surgery on the remnant. AB - The residual aneurysm rate is reported between 3,8% and 21% in the cases followed after intracranial aneurysm surgery. In the formation of the residual aneurysm, the risk factors include such structural characteristics as the size and lobulation of the aneurysm, posterior circulation, para-ophthalmic localisation and intra-operative rupture. The rates and causes of postoperative residual aneurysms were analyzed in 186 intracranial aneurysm of 160 patients, including the possible effects of temporary clipping on the residual rates. The entire series demonstrated a residual rate of 7%. It was found higher in the large lobulating aneurysms and intra-operative rupture. The residual rate considerably decreased to 4,2% in the aneurysms with temporary clipping. The determination of residual aneurysms, identification of any risk factors and elimination of recoverable factors would allow improvement of surgical results in the treatment of the intracranial aneurysms in the future. Moreover we believe that these will be useful in development of the indications for alternative treatment methods. PMID- 11862513 TI - Basilar vasospasm following spontaneous and traumatic subarachnoid haemorrhage: clinical implications. AB - BACKGROUND: Cerebral vasospasm has been commonly described following subarachnoid haemorrhage (SAH) though its impact on neurological outcome, especially in head trauma, has not been yet elucidated. The purpose of this study was to monitor and correlate neurological condition and flow velocities (FVs) in the arteries of the brain after SAH and more particularly to investigate the influence of basilar artery (BA) vasospasm on neurological outcome. METHODS: Daily transcranial Doppler (TCD) evaluations were conducted in 116 consecutive patients with subarachnoid haemorrhage. SAH was of traumatic origin (tSAH) in 59 patients and spontaneous (sSAH) in 57 patients. Vasospasm in the MCA and ACA was defined by a mean FV exceeding 120 cm/s and three times the mean FV of the ipsilateral ICA. Basilar artery (BA) vasospasm was defined as moderate whenever the FV was higher than 60 cm/s and severe above 85 cm/s. FINDINGS: Sixty-two patients (53.4%) had elevated FVs in the BA, among these 34 (29.3%) had FVs above 85 cm/s. Basilar vasospasm was significantly more common in tSAH (59.7%) than in sSAH (40.3%, P=0.041). In patients with moderate and severe BA vasospasm, FVs in the BA increased on the third day after admission and remained elevated for a week before returning to normal value by the end of the second week. This elevation in BA FVs in patients with BA vasospasm was followed by a significant and progressive worsening in the neurological condition at the end of the first week. Permanent neurological deficit was associated with elevated BA FVs consistent with moderate BA vasospasm whereas patients who remained in persistent vegetative state, had FVs consistent with severe BA vasospasm (P=0.00019). INTERPRETATION: The present results further support that BA vasospasm may act as an independent factor of ischaemic brain damage following SAH, especially in head trauma. PMID- 11862515 TI - Imbalance between apostain expression and proliferative index can predict survival in primary glioblastoma. AB - BACKGROUND: Cell proliferation and cell death are opposing processes in tumour growth, with tumour progression reflecting the balance between proliferating and apoptotic cells. The purpose of the present study is to verify the hypothesis that an imbalance between apoptosis and proliferation can predict survival in patients with primary glioblastoma. METHODS: After the immunohistochemical study of Apostain and MIB-1 expression, the index of apoptosis (AI), the index of proliferation (PI), and the ratio AI/PI was recorded for each tumour specimen, in a series of 32 primary glioblastomas. Studies of correlation between AI and PI, between AI and survival, between PI and survival, and between the ratio AI/PI and survival, were performed using the Spearman rank correlation test. Furthermore, a comparative study of survival was performed for subgroups of patients with ratio AI/PI greater or lesser than 1, using the log rank test. FINDINGS: In the present series, values of AI and PI showed a wide distribution, with a mean +/- SD of 8.16 +/- 7.2, and of 12.69 +/- 21.1, respectively. The values for the ratio AI/PI ranged between 0.01 and 6.03 (mean +/- SD: 1.44 +/- 1.60). Statistical study failed to obtain correlation between AI and PI. Survival of patients not correlated with AI neither with PI. The ratio between AI and PI did not correlate with survival either. Nevertheless, when survival for the subgroups of patients showing a ratio AI/PI greater or lesser than 1 was compared, a significant difference was found (p: 0.02). Survival ranged between 50 and 81 weeks (mean of 58.5 +/- 11.05 weeks) for the 12 cases showing a ratio AI/PI greater than 1, and it ranged between 8 and 85 weeks (mean: 38.20 +/- 25.37 weeks) for the 20 cases showing a ratio AI/PI lesser than 1. INTERPRETATION: Our present results suggest that a clear imbalance between cell proliferation and apoptosis can predict outcome in patients operated on for a primary glioblastoma. PMID- 11862514 TI - Cerebral germinoma with hemiatrophy of the brain: report of three cases. AB - BACKGROUND: The authors report three cases of cerebral germinoma that occurred in young adults with unusual presentation. METHOD: All three patients presented with hemiparesis and were treated at Seoul National University. A histological diagnosis of germinoma was made by a stereotactic biopsy in all three cases. FINDINGS: Magnetic resonance (MR) images showed that their tumors were located in the internal capsule and thalamus, and were associated with ipsilateral cerebral hemisphere and brain stem atrophy. The hemiparesis slowly progressed and this was accompanied by a haemorrhagic cyst in each patient. INTERPRETATION: Clinical diagnosis was not easy because of the unusual clinical presentations and atypical MR imaging findings. It is suggested that cerebral germinoma should be included in the differential diagnosis of a haemorrhagic mass which is associated with cerebral atrophy in the thalamus, basal ganglia, or internal capsule, especially in adolescents or young adults. PMID- 11862516 TI - (201)Tl-SPECT, (1)H-MRS, and MIB-1 labeling index of central neurocytomas: three case reports. AB - BACKGROUND: The proliferative activity and metabolic features of three central neurocytomas were investigated using the findings of thallium-201 single photon emission computed tomography ((201)Tl-SPECT) and proton magnetic resonance spectroscopy ((1)H-MRS), and the MIB-1 labeling index (MIB-1 LI). METHOD: The early and delayed (201)Tl indices were calculated as the ratio of tumour to normal brain tissue uptake by (201)Tl-SPECT. In vivo single-voxel (1)H-MRS was performed with echo time of 272 msec to evaluate the metabolites including choline (Cho), N-acetyl aspartate (NAA) and creatine/phosphocreatine (Cre). An external standard reference was used to semiquantitate each metabolite. MIB-1 LI was determined in the surgical specimens. FINDINGS: The MIB-1 LI was 0.5%, 1.2%, and 7.5% in an atypical central neurocytoma without intraventricular extension. Significant (201)Tl uptake was observed on delayed images in all three central neurocytomas. (1)H-MRS showed the high Cho peaks relative to the NAA and Cre peak. The signal at 3.55 ppm, which may be due to inositol or glycine, was observed in one central neurocytoma. INTERPRETATION: Both (201)Tl-SPECT and (1)H MRS did not reflect the proliferative potential of central neurocytomas. PMID- 11862517 TI - Morphometric measurements of the cranium in patients with Chiari type I malformation and comparison with the normal population. AB - BACKGROUND: To determine the degree of development of the posterior fossa and signs of occipital dysplasia in patients with Chiari type I malformation by morphometric measurements. METHODS: In 22 patients with Chiari type I malformation, distance, surface area and angle values reflecting the degree of development of the posterior fossa were measured and compared with the measurements of 21 normal subjects. FINDINGS: In patients with Chiari type I malformation, the depth of the posterior fossa, the length of the clivus reflecting development of the basi-occiput and Klaus' index were significantly shorter than in normal subjects (p<0.001, p=0.007, and p<0.001, respectively). The ratios of the depth of the posterior fossa to the height of the supratentorial region and Twining's line reflecting anteroposterior distance of the posterior fossa were also significantly smaller in the Chiari group (p<0.001 for both). In sagittal section, the surface area of the bony part of the posterior fossa and the ratio of this area to the area of the supratentorial region were significantly smaller than in normal subjects (p=0.038 and p=0.002, respectively). The angle measurements of the cranial base (basal angle, Boogard's angle and nasion-basion-opisthion angle) showed that there was an evident tendency for platibasia in the Chiari group (p=0.04, p=0.004, p<0.001, respectively). In addition, it was shown by measuring tentorium-twining's line angle that the tentorium was steeper in the Chiari group than normal subjects. INTERPRETATION: These results support the opinion, which claims the existence of underdevelopment of the occipital bone and posterior fossa in patients with Chiari type I malformation. PMID- 11862518 TI - Experimental research photodynamic effects in perifocal, oedematous brain tissue. AB - BACKGROUND: Photodynamic therapy (PDT) has been under discussion as additional treatment option for malignant gliomas. However, damage not only to tumour tissue but also to normal brain has been demonstrated. The mechanisms of this unwanted side effect have not yet been clearly identified. Spreading of photosensitiser with oedema after disruption of the blood-brain-barrier and potential sensitisation of normal tissue has been found previously. The present study investigates the time- and dose-dependency of normal tissue damage to photodynamic therapy using Photofrin II after disruption of the blood-brain barrier. METHODS: Male wistar rats anaesthetised with chloral hydrate were subjected to focal, cerebral cold lesions. Simultaneously, Photofrin II (PFII) was injected (2,5 or 5 mg/kg b.w.). Laser irradiation (630 nm) was performed after 4 h, 12 h and 24 h with varying light doses. Control groups were subjected to focal cold lesion alone, cold lesion with laser irradiation, PFII followed by laser irradiation, or laser irradiation alone (n=6 all groups). 24 h later, brains were removed for assessment of necrosis in coronal sections. FINDINGS: Light dose had a significant impact on the extent of necrosis. Compared to control animals (lesion only: 0.84 +/- 0.2 mm2; lesion and irradiation alone: 0.7 +/- 0.3 mm2), the area of necrosis was increased to 2.8 +/- 0.5 (50 J/cm2), 3.5 +/- 1,1 (100 J/cm2) and 4.3 +/- 0.7 mm2 (200 J/cm2, 5 mg/kg b.w.; p<0.01). This effect was time-dependent. Maximal necrosis (6.3 +/- 1,6 mm2) was observed when brains were irradiated 12 h after PFII injection, with less necrosis occurring at 24 h (2.8 +/- 0.4 mm2, p<0.01). Reducing sensitiser dose to 2.5 mg/kg b.w. resulted in a reduction of necrosis (2.09 +/- 0.2 mm2, p<0.05). INTERPRETATIONS: Damage to oedematous tissue after photodynamic therapy using i.v. PFII and laser light at 630 nm depends on laser dose, sensitiser dose and the time point of laser irradiation. The time point of PDT should be considered to prevent unwanted tissue reactions. In the clinical setting however, defined damage to peritumoural tissue may be advantageous. This should be achievable by optimised timing and dosage of photodynamic therapy. PMID- 11862519 TI - Regeneration of auditory nerve following complete sectioning and intrathecal application of the IN-1 antibody. AB - The cochlear nerve of adult Lewis rats was following microsurgical exposure in the cerebellopontine angle (CPA). The lesions completely interrupted the auditory nerve axons at the lesion site producing ipsilateral deafness in all animals. The rats were then treated with a recombinant Fab fragment of the antibody IN-1 against nerve growth inhibitory proteins for one to two weeks. An age-matched control group of rats was treated with unspecific mouse IgG antibody. Because the cochlear nerve lesions resulted in significant neuronal apoptosis of spiral ganglion cells, neurotrophin-3 (NT-3) was applied to the lesion site immediately post-injury in some rats. Electrophysiological studies were carried out by recording the brainstem auditory evoked potentials (BAEP) before and immediately after the lesion, and at regular intervals up to 2 months after injury. Cochlear nerve fibres were anterogradely traced by horseradish peroxidase (HRP) or biotinylated dextran amine (BDA) injected into the spiral ganglion. The results achieved in this study were consistent with the following conclusions: 1) transection of the adult rat cochlear nerve at the CPA results in functional deafness, disappearance of BAEP, apoptosis of parent axotomized neurons of the spiral ganglion, and interruption of labelled axons close to the lesion site; 2) NT-3 is able to partially rescue axotomized neurons of the spiral ganglion; 3) injured cochlear nerve fibres show a limited spontaneous sprouting and regrowth response which does not lead to BAEP recovery; 4) intrathecal treatment with IN-1 directed against myelin-associated neurite growth inhibitory proteins promotes significant elongation of the injured fibres; and 5) the regenerating fibres seem to navigate to correct targets, and be able to establish synaptic connections for functional recovery as depicted by BAEP examinations. PMID- 11862520 TI - Alteration of ischemic reperfusion injury in the rat neocortex by a potent antioxidant mexiletine. AB - The mechanisms by which mexiletine exerts its effects in increasing myocardial circulation, and smooth muscle perfusion and alleviating diabetic neuropathic pain have been widely discussed. The purpose of this study was to examine the protective effect of this compound in ischemia/reperfusion-induced cerebral injury following middle cerebral artery occlusion in Sprague-Dawley rats. Blood flow to the left cerebral hemisphere of the animals was interrupted by occluding the left cerebral artery and both carotid arteries simultaneously for 3 hrs. These animals were assigned to one of ten groups and divided into treatment group and pretreatment group; 1) control treatment group (n=8); 2) vehicle treatment group (n=8); 3) lower dose mexiletine (400 microg/kg) treatment group (n=8); 4) medium dose mexiletine (800 microg/kg) treatment group (n=8); 5) high dose mexiletine (2 mg/kg) treatment group (n=8); 6) control pretreatment group (n=8); 7) vehicle pretreatment group (n=8); 8) lower dose mexiletine (400 microg/kg) pretreatment group (n=8); 9) medium dose mexiletine (800 microg/kg) pretreatment group (n=8); and 10) high dose mexiletine (2 mg/kg) pretreatment group (n=8). The volume of cerebral infarction was measured in serial brain sections stained with triphenyltetrazolium chloride (TTC). Tissue infarction volume and tissue edema were estimated for each animal. The volume of cerebral infarction was significantly decreased in rats pretreated with mexiletine, and the ratio of tissue edema was also decreased as the dose of mexiletine increased. These results demonstrate that mexiletine, an anti-arrhythmic and use-dependent Na+ channel blocker, has protective effects in stroke at concentrations sufficient to confer significant protection, as measured by the volume of infarction and brain edema index in a model of focal, neocortical ischemia in Sprague-Dawley rats. PMID- 11862521 TI - Influence of aging on blood-brain barrier permeability and free radical formation following experimental brain cold injury. AB - The aim of this study is to investigate the effects of experimental cold brain injury on blood-brain barrier integrity, on brain oedema formation, and on lipid peroxidation and to compare the results between the aged and young rats. Cold brain injury was used to create a standard model of brain trauma in old and young rats. Disruption of the blood-brain barrier was analyzed by Evans blue method. The values of cerebral water content were calculated by using the fresh and dry weights of the cerebral hemispheres. Lipid peroxidation was assessed by measuring the tissue content of malonyl dialdehyde. Blood-brain barrier was destroyed significantly in young and old rats, but it was more severe in old rats. Accordingly, cerebral water content was increased in both groups, however this increase was significantly more prominent in old rats. No significant difference was found on malonyl dialdehyde levels between young and old rats. The blood brain barrier was more easily disrupted in old rats, and this was supposed to be the basic event causing more secondary damage. PMID- 11862522 TI - Supratentorial leptomeningeal metastasis of a medulloblastoma without cerebellar tumor recurrence. AB - The authors report on a 15-year-old girl presenting with headache and nausea. Cranial magnetic resonance imaging (MRI) showed a diffuse leptomeningeal contrast enhancement 5 years after resection of a posterior fossa medulloblastoma followed by radio- and chemotherapy. A left frontal biopsy revealed tumor spread. Histopathological findings of the tumor were identical to those obtained from the original medulloblastoma. To the authors' knowledge this is the first report of late supratentorial leptomeningeal metastasis from a medulloblastoma while no local recurrence or spinal leptomeningeal dissemination had occurred. PMID- 11862523 TI - Cervical myelopathy due to gouty tophi in the intervertebral disc space. AB - Gout, like diabetes mellitus, is a common metabolic disorder. Typically affecting the distal joint of the appendicular skeleton, its occurrence in the spine is rare. We report the case of a 68-year-old male with a long history of diabetes mellitus and hyperuricemic gout. Neck pain developed over two weeks with subsequent quadriparesis, with concomitant subcutaneous deposition of gouty tophi in the right elbow. Magnetic resonance image of the cervical spine revealed multiple segmental narrowing of the thecal sac at the C3-6 levels due to hypertrophic spurs and bulging discs. Anterior discectomies of C3-4 and C4-5 were performed, with a chalky-white, granular material noted in the C4-5 disc space. Histological examination of the surgical specimen revealed deposits of needle like crystals surrounded by histiocytes and multinucleated giant cells, with the appearance compatible with gout. The patient was ambulatory with the assistance of a walking frame six months after the operation. We emphasize that gouty tophi can be deposited in the spine over a relatively short time, subsequently precipitating a variety of symptoms, from pain to cord compression. The regular administration of antihyperuricemia drug treatment for hyperuricemic gout is necessary to prevent this deposition. If neurological defects are found, surgical decompression can provide satisfactory results. PMID- 11862524 TI - A case of pineal region meningioma without dural attachment, presented with bilateral hearing impairment. PMID- 11862525 TI - Spinal epidural lipomatosis associated with pituitary macroprolactinoma. PMID- 11862526 TI - Active vitamin D and its analogs as drugs for the treatment of osteoporosis: advantages and problems. PMID- 11862527 TI - Development of a synthetic peptide-based tartrate-resistant acid phosphatase radioimmunoassay for the measurement of bone resorption in rat serum. AB - Selective markers of bone turnover provide a convenient and reproducible alternative to the complex and expensive histochemical techniques used commonly to study the effect of pharmacological agents and the pathogenesis of bone disease in the ovariectomized (OVX) rat model. One marker, which has been specifically linked to terminally differentiated osteoclasts and, thus, provides useful insight at cellular levels, is type-5 tartrate-resistant acid phosphatase (TRACP). We describe the development of a TRACP radioimmunoassay (RIA), which requires synthetic peptide for antibody development. To develop the RIA, polyclonal antibodies were generated in goats against a synthetic peptide, DPSVRHQRKCY, corresponding to amino acid residues 267-275 of the rat type-5 TRACP sequence. In the RIA, 50 microl of rat serum, 100 microl of goat anti-TRACP antibodies, and 100 microl of tracer were incubated overnight. The antibody-bound fraction was separated, counted, and unknown values were calculated by comparison with the peptide calibrator. Rat serum shows parallelism with the synthetic peptide calibrator used in the RIA. The sensitivity of the RIA was 24.7 microg/l, and the measuring range was 19-2476 microg/l. The average intra-assay coefficients of variation for (CV) two controls were less than 7%. The average dilution and spike recoveries were 107% and 87%, respectively. We applied our peptide-based RIA to study bone resorption in an OVX rat model. TRACP concentrations in serum in 12-week-old OVX Sprague Dawley rats were 14%-22% (P < 0.05) higher than those in the sham-operated rats, and TRACP concentrations in OVX rats treated with estradiol were 24%-32% lower (P < 0.01) than those in the vehicle-treated OVX group. Similarly, as compared with those in OVX rats, TRACP concentrations decreased to those of sham levels in OVX rats receiving 10 microg/kg per day of alendronate for 10 days. In addition, the TRACP levels determined by RIA showed a significant correlation with serum C-telopeptide (type I collagen) concentrations (r = 0.56; P < 0.001) measured by an enzyme-linked immunosorbent assay (ELISA) developed earlier for the rat model. In conclusion, we have developed a TRACP RIA that could be used to monitor the rate of bone resorption in the rat model. PMID- 11862528 TI - Selective synthesis of bone morphogenetic proteins-1, -3, -4 and bone sialoprotein may be important for osteoinduction by Saos-2 cells. AB - An ability to induce new bone formation at a required site would represent a considerable advance in bone repair and tissue engineering. It has been shown that the healing of critical-size bone defects in rats can be augmented by extracts of Saos-2 cells. These human osteosarcoma cells uniquely contain a bone inducing activity, whereas other human osteosarcoma cells, e.g., U-2 OS cells, cannot replicate the osteoinductive capacity. To understand the necessary components of the Saos-2 bone-inducing activity, this study compared osteoinductive Saos-2 cells with non-osteoinductive U-2 OS cells with respect to the synthesis of bone morphogenetic proteins (BMPs)-1, -2, -3, -4, -5, -6, and -7 and the non-collagenous matrix proteins bone sialoprotein (BSP), osteonectin (ON), osteopontin (OPN), and osteocalcin (OC). The main differences were abundant synthesis of BMP-1/tolloid, BMP-3, -4, and BSP by Saos-2 cells, but absence or reduced synthesis in U-2 OS cells. BMP-2 and -7 were present in low amounts in both cell types, while BMP-5 and -6 were more abundant in U-2 OS cells, suggesting that these BMPs were of lesser importance for the osteoinductivity of Saos-2 cells. However, a relatively high expression of BMP-3 and -4, together with BMP-1/tolloid, may be important for the osteoinductive capacity of Saos-2 cells. The inability of U2-OS cells to induce bone, despite expressing most of the BMPs, may be due to an insufficiency of tolloid, BMP-3 or -4, BSP, and/or other unknown factors. A better understanding of the necessary components of the Saos-2 cell bone-inducing agent may, in future, lead to clinically useful Saos-2 cell products for bone repair and tissue engineering. PMID- 11862529 TI - Human PTH (1-34) induces longitudinal bone growth in rats. AB - The growth plate is a specialized structure that is responsible for longitudinal bone growth (LGR). Growth plate organization is altered with loading in rats. Parathyroid hormone (PTH) is known to induce mitogenic effect on chondrocytes in vitro. Type I PTH/PTH related peptide (rP) receptor is expressed in growth plate cartilage in rats. We therefore investigated the effect of PTH administration on the organization and longitudinal growth rate of the growth plate in rats. We also investigated the effect of PTH on the changes induced by unloading in the organization and growth of the growth plate. Thirty 6-week-old and 30 15-week-old male Sprague-Dawley rats were randomly assigned to five groups (n = 6 per group), i.e., basal controls, control (i.e., normally loaded), PTH-treated control (i.e., PTH-treated under normal loading), unloaded, and PTH-treated under unloading. PTH treated animals received human PTH (1-34) at a dose of 80 microg/kg per day five times per week for 3 weeks, for the duration of unloading. In young loaded rats treated with the systemic administration of PTH, growth plate thickness, chondrocyte number, and LGR were increased in the proximal tibiae compared with findings in young loaded rats without PTH administration. Hindlimb unloading induced a reduction in growth plate thickness, chondrocyte number, and LGR. In young rats, systemic administration of PTH partly prevented these changes induced by unloading. These preventive effects of PTH were observed only in young rats; not in adult rats. These results show that the systemic administration of PTH stimulates longitudinal bone growth, and diminishes the reduction in growth plate growth induced by unloading in young rats. PMID- 11862530 TI - Osteoporosis following chronic constriction injury of sciatic nerve in rats. AB - Osteopathic changes sometimes occur in patients with complex regional pain syndrome (reflex sympathetic dystrophy and causalgia). We aimed to investigate whether such osteopathic changes occurred in rats with chronic constriction injury (CCI) of the sciatic nerve. A CCI of the sciatic nerve was established in a unilateral hind limb in 39 adult Sprague-Dawley rats, which were killed 1, 2, 3, 5, or 7 weeks after the CCI procedure. Bone mineral content (BMC) and bone mineral density (BMD) in extracted tibial bones were measured using a dual-energy X-ray absorptiometer, and the number of osteoclasts in the metaphyseal regions was counted by the use of tartrate-resistant acid phosphate (TRAP) staining. BMC was significantly decreased, compared with that of the contralateral side, 1 to 7 weeks after CCI, and BMD was decreased 2 to 7 weeks after the procedure in the ipsilateral tibial bones, compared with BMD in the contralateral bones. The number of TRAP-positive multinucleated osteoclasts in the ipsilateral bones was significantly increased at 2, 3, and 5 weeks after the CCI, when compared with the number of these osteoclasts in the contralateral bones. The results of the present study demonstrate that osteopathic changes are associated with chronic constrictive injury of the sciatic nerve. PMID- 11862531 TI - A selective inhibitor of matrix metalloproteinases inhibits the migration of isolated osteoclasts by increasing the life span of podosomes. AB - The osteoclast is a unique cell that cycles between bone resorption and migration. In this study, we used KB-R7785, an inhibitor of matrix metalloproteinases (MMPs), to investigate the role of MMPs in this cycle. Osteoclasts prepared from neonatal rabbits were processed to measure: (1) migration on hydroxyapatite (HA)-coated dishes, (2) the formation of an actin band, which is a large collection of podosomes, (3) the life span of podosomes, and (4) lacunar resorption on ivory slices. KB-R7785 significantly decreased the area of the tracks formed by osteoclasts on HA-coated dishes. The percentage of actin/podosome band formation, expressed as: (total length of the actin band/circumference of the osteoclast) x 100, was significantly increased after the addition of KB-R7785, which implies that podosome disassembly in osteoclasts was decreased by the inhibition of MMPs. Time-lapse cinemicrography demonstrated that the addition of KB-R7785 increased the life span of podosomes. Several indexes (number of resorption pits, total resorption area, and mean resorption area) of osteoclast resorption activity were significantly decreased by KB-R7785, while the average depth of resorption was increased. These results indicate that decreased podosome disassembly caused by an MMP inhibitor suppresses the migration of osteoclasts and, ultimately, inhibits osteoclastic bone resorption. PMID- 11862532 TI - A study of bone mineral density and physical growth in very low birth-weight infants after their discharge from hospital. AB - The lumbar spinal bone mineral density (BMD) of very low birth-weight (VLBW) infants was determined by dual-energy X-ray absorptiometry, and its relationship with physical growth was analyzed. The influence of birth-related factors on changes in BMD after discharge from the hospital were also investigated. The BMD increased rapidly until the age of 2 years, in association with improvements in nutritional status and the increase in physical growth after discharge. The Z score, which indicates the degree of attainment of the age-specific standard BMD, was determined to be almost 90% at the age of around 2 years and older. Significant increases in the BMD continued to be noted after the age of 2 years, along with increases in body weight and body height. At the age-adjusted value at less than 1 year, however, the Z score tended to be low when the birth weight (<1000 g) and birth height (<36 cm) were low (P < 0.02, respectively), suggesting that the birth weight and birth height influence the BMD at this age. These results indicate that the condition at birth and nutritional management during hospitalization affect the BMD soon after discharge, but that the nutritional condition after discharge becomes a more important factor at the age of 2 years and older. PMID- 11862533 TI - Ultrasound assessment of tibial cortical bone acquisition in Japanese children and adolescents. AB - The purpose of the present study was to evaluate the normal process of cortical bone acquisition during childhood and adolescence, and the relationship between speed of sound (SOS), measured by the Sound Scan 2000 system, and linear growth. A total of 1689 healthy Japanese children and adolescents (862 males and 827 females, aged 7-19 years) were enrolled in the study. SOS (m/s) was measured at the right tibial midshaft and the standard SOS values in the children and adolescents were generated. Various growth parameters were also measured. SOS of tibia increased significantly with age in both males and females (P < 0.001 each). In both sexes, a spurt in SOS was noted 1 year after the standard age at which Japanese peak height velocity (PHV; cm/year) occurs, and SOS increased markedly after the age at which the length of tibia reached a maximum. Multiple regression equations for SOS were generated as a function of various growth parameters; for males, SOS = 3047 + 6.2 x height (cm) + 2.1 x weight (kg) - 9.8 x length of tibia (cm) (R2 = 0.50; P < 0.001) and for females, SOS = 2713 + 10.3 x height (cm) + 1.8 x weight (kg) - 15.5 x length of tibia (cm) (R2 = 0.49; P < 0.001). In both sexes, SOS correlated positively with body height and weight, but negatively with length of tibia. Our results indicate that SOS is quite useful for evaluating cortical bone status in children and adolescents; the results of these measurements may provide an explanation for the relative weakness of the mechanical properties of the bone during childhood and adolescence. PMID- 11862534 TI - Effect of surgical menopause and Paget's disease of bone on the isomerization of type I collagen carboxyterminal telopeptide: evolution after antiresorptive therapy. AB - The objective of this study was to analyze the effect of surgical menopause and Paget's disease of bone, as well as the influence of therapy, on the isomerization of the carboxyterminal telopeptide of type I collagen (CTX). Fourteen women who had undergone surgical menopause and had begun hormone replacement therapy (HRT) after surgery were recruited. Results for these women were compared with those of 29 patients with Paget's disease of bone treated with tiludronate (400 mg/day) for 3 months, and with those of a group of 21 healthy premenopausal women (control group I). In addition, 14 healthy individuals with an age range similar to that of the pagetic patients (control group II) were included in the study. Urine samples were analyzed for levels of nonisomerized and beta-isomerized CTX (alpha-CTX and beta-CTX). Biochemical determinations were performed 3 months after surgical menopause and after 3 and 9 months of HRT, and at baseline, and 1 and 6 months after tiludronate treatment in the pagetic patients. The average levels of alpha-CTX and beta-CTX were higher in patients than in controls. In patients after surgical menopause, because of their greater increase of beta-CTX, the alpha-CTX/beta-CTX ratio was lower than that of control group I (0.881 +/- 0.3 vs 1.515 +/- 0.8; P < 0.05). In contrast, at baseline, pagetic patients showed marked increases in alpha-CTX levels, resulting in a higher alpha-CTX/beta-CTX ratio than that of control group II (2.879 +/- 1.3 vs 0.96 +/- 0.25; P < 0.0001). The average percent decrease in both markers after therapy was similar in both conditions (-60% for alpha-CTX and -44% for beta-CTX after 3 months of HRT in the surgical menopause group, vs -66% for alpha-CTX and 41% for beta-CTX in the pagetic group, 1 month after finishing tiludronate therapy; P, NS), resulting in a significant decrease of the alpha-CTX/beta-CTX ratio in pagetic patients (2.879 +/- 1.3 vs. 1.614 +/- 0.8; P < 0.001). In conclusion, surgical menopause is associated with a decrease in the urinary alpha CTX/beta-CTX ratio because of the higher increase in the beta-CTX level after menopause. Pagetic patients show an increase in this ratio, compared with the control value, and the ratio decreases after bisphosphonate treatment. The response to therapy was similar in both conditions, with a comparable decrease of both markers. These findings show how bone markers may contribute to the understanding of pathophysiologic mechanisms in bone diseases. PMID- 11862536 TI - An evaluation of structural models for the photosynthetic water-oxidizing complex derived from spectroscopic and X-ray diffraction signatures. AB - Four of the five intermediate oxidation states (S-states) in the catalytic cycle of water oxidation used by O2-evolving photoautotrophs have been previously characterized by EPR and/or ENDOR spectroscopy, with the first reports for the S0, S1, and S3 states available in just the last three years. The first electron density map of the Mn cluster derived from X-ray diffraction measurements of single crystals of photosystem II at 3.8-4.2 A resolution has also appeared this year. This wealth of new information has provided significant insight into the structure of the inorganic core (Mn4OxCa1Cl1-2), the Mn oxidation states, and the location and function of the essential Ca2+ cofactor within the water-oxidizing complex (WOC). We summarize these advances and provide a unified interpretation of debated structural proposals and Mn oxidation states, based on an integrated analysis of the published data, particularly from Mn X-ray absorption spectroscopy (XAS) and EPR/ENDOR data. Only three magnetic spin-exchange models for the inter-manganese interactions are possible from consideration of the EPR data for the S0, S1, S2 and S(-N) (NO-reduced) states. These models fall into one of three types denoted butterfly, funnel, or tetrahedron. A revised set of eight allowed chemical structures for the Mn4Ox core can be deduced that are shown to be consistent with both EPR and XAS. The popular "dimer-of-dimers" structural model is not compatible with the possible structural candidates. EPR data have identified two inter-manganese couplings that are sensitive to the S-state, suggesting two possible bridging sites for substrate water molecules. Spin densities derived from 55Mn hyperfine data together with Mn K-edge energies from Ca-depleted samples provide an internally consistent assignment for the Mn oxidation states of Mn4(3III,IV) for the S2 state. EPR and XAS data also provide a consistent picture, locating Ca2+ as an integral part of the inorganic core, probably via shared bridging ligands with Mn (aqua/hydroxo/carboxylato/chloro). XAS data reveal that the Ca2+ cofactor increases the Mn(1s-->4p) transition energy by 0.6-1 eV with minimal structural perturbation versus the Ca-depleted WOC. Thus, calcium binding appears to increase the Mn-ligand covalency by increasing electron transfer from shared ligands to Mn, suggesting a direct role for Ca2+ in substrate water oxidation. Consideration of both the XAS and the EPR data, together with reactivity studies on two model complexes that evolve O2, suggest two favored structure types as feasible models for the reactive S4 state that is precursor to the O2 evolution step. These are a calcium-capped "cuboidal" core and a calcium-capped "funnel" core. PMID- 11862537 TI - A modeling study of the interaction and electron transfer between cytochrome b5 and some oxidized haemoglobins. AB - Using Brownian motion simulations we have studied the formation of docked complexes of reduced cytochrome b5 and oxidized haemoglobin. Our results indicate that the presence of molecular electrostatic fields has a significant role to play in the formation of these complexes. In contrast to previous modeling studies on this system, we clearly identify electron transfer within an ensemble of similarly docked complexes rather than the formation of a single complex. Docking involves a number of acidic residues surrounding the exposed haem edge of cytochrome b5 and a set of basic residues surrounding the exposed haem edge of the globins. Although amino acids from the partner globin proteins are involved to a small extent in the binding of some of the complexes, the reactivity of any particular globin is essentially independent of the nature of its partner globin chain within the haemoglobin molecule. Comparison of results from adult and embryonic haemoglobins indicates a significant difference in complex formation. Application of electron tunneling analysis to the complexes allows us to predict the rates of electron transfer within each ensemble of complexes. These data provide a theoretical insight into the important process of re-reduction of oxidized haemoglobins as well as explaining the general inability to produce crystalline forms of many docked electron transfer complexes. PMID- 11862538 TI - Reaction of peroxynitrite with carbon dioxide: intermediates and determination of the yield of CO3*- and NO2*. AB - CO2 catalyses the isomerization of the biological toxin ONOO- to NO3- via an intermediate, presumably ONOOCO2-, which has an absorption maximum near 650 nm. The reflection spectrum of solid NMe4+ ONOO- exposed to CO2 shows a similar band near 650 nm; this absorption decays over minutes. Stopped-flow experiments in which CO2 solutions were mixed with alkaline ONOO- solutions indicate the formation of at least one intermediate. The initial absorption at 302 nm is less than that of ONOO-, which indicates that reactions take place within the mixing time, and this absorption is dependent (but not linearly) on the ONOO- and CO2 concentrations. We found that reaction of peroxynitrite with carbon dioxide forms some trioxocarbonate(*1-) (CO3*-) and nitrogen dioxide (NO2*) radicals via homolysis of the O-O bond in ONOOCO2-. We determined the extent of radical formation by mixing peroxynitrite, carbon dioxide and nitrogen monoxide. The later reacts with CO3*- and NO2* radicals to form, effectively, three NO2- per homolysis; ONOOCO2- that does not undergo homolysis yields NO3- and CO2. Based on the NO3- and NO2- analyses, the extent of conversion to NO3- is 96 +/- 1% and that of homolysis is 3 +/- 1%, respectively, significantly less than that reported in the literature. PMID- 11862539 TI - The Fe-only nitrogenase from Rhodobacter capsulatus: identification of the cofactor, an unusual, high-nuclearity iron-sulfur cluster, by Fe K-edge EXAFS and 57Fe Mossbauer spectroscopy. AB - Samples of the dithionite-reduced FeFe protein (the dinitrogenase component of the Fe-only nitrogenase) from Rhodobacter capsulatus have been investigated by 57Fe Mossbauer spectroscopy and by Fe and Zn EXAFS as well as XANES spectroscopy. The analyses were performed on the basis of data known for the FeMo cofactor and the P cluster of Mo nitrogenases. The prominent Fourier transform peaks of the Fe K-edge spectrum are assigned to Fe-S and Fe-Fe interactions at distances of 2.29 A and 2.63 A, respectively. A significant contribution to the Fe EXAFS must be assigned to an Fe backscatterer shell at 3.68 A, which is an unprecedented feature of the trigonal prismatic arrangement of iron atoms found in the FeMo cofactor of nitrogenase MoFe protein crystal structures. Additional Fe...Fe interactions at 2.92 A and 4.05 A clearly indicate that the principal geometry of the P cluster is also conserved. Mossbauer spectra of 57Fe-enriched FeFe protein preparations were recorded at 77 K (20 mT) and 4.2 K (20 mT, 6.2 T), whereby the 4.2 K high-field spectrum clearly demonstrates that the cofactor of the Fe-only nitrogenase (FeFe cofactor) is diamagnetic in the dithionite-reduced ("as isolated") state. The evaluation of the 77 K spectrum is in agreement with the assumption that this cofactor contains eight Fe atoms. In the literature, several genetic and biochemical lines of evidence are presented pointing to a significant structural similarity of the FeFe, the FeMo and and the FeV cofactors. The data reported here provide the first spectroscopic evidence for a structural homology of the FeFe cofactor to the heterometal-containing cofactors, thus substantiating that the FeFe cofactor is the largest iron-sulfur cluster so far found in nature. PMID- 11862540 TI - 1H NMR investigations of the molecular nature of low-molecular-mass calcium ions in biofluids. AB - High-resolution 1H NMR spectroscopy was employed to explore the complexation of Ca2+ by low-molecular-mass biomolecules in human saliva. The results acquired revealed that the organic acid anion (OAA) citrate acts as a powerful oxygen donor chelator for salivary Ca2+, and accurate determination of its resonances' frequencies and spin-system pattern could be successfully utilized to determine its degree of saturation with this metal ion. Computer modelling studies demonstrated that the OAA lactate is the only competing salivary Ca2+ complexant available. Moreover, the Ca2+-complexation status of salivary citrate is substantially modified by dentifrice-mediated elevations in its concentration. 1H NMR analysis was also applied to determinations of the Ca2+ saturation status of citrate in a variety of alternative biofluids and the biochemical significance of these results is discussed. PMID- 11862541 TI - Relaxometric evaluation of novel manganese(II) complexes for application as contrast agents in magnetic resonance imaging. AB - Three novel Mn(II) complexes bearing benzyloxymethyl functionalities are reported and their ability to enhance water (1H and 17O) relaxation times is investigated in detail. Two of them contain one coordinated water molecule and display relaxivity values only slightly smaller than those shown by the most clinically used contrast agents (e.g. [Gd(DTPA)(H2O)]2-). Moreover, in these Mn(II) chelates the exchange rate of the coordinated water is ca. one order of magnitude higher if compared to the exchange rates previously reported for Gd(III) complexes with octadentate ligands. The occurrence of such fast exchange rates of the coordinated water is exploited in the formation of macromolecular adducts with human serum albumin to attain systems displaying relaxivity values in the upper range of those so far reported for analogous Gd(III) systems. These results strongly support the view that Mn(II) complexes, in spite of the lower effective magnetic moment, can be considered as viable alternatives to the currently used Gd(III) complexes as contrast agents for MRI applications. PMID- 11862542 TI - Development of new insulinomimetic zinc(II) picolinate complexes with a Zn(N2O2) coordination mode: structure characterization, in vitro, and in vivo studies. AB - Three zinc(II) complexes of picolinic acid and its derivatives with a Zn(N2O2) coordination mode were prepared and evaluated for their insulinomimetic activities by in vitro and in vivo studies. By introducing an electron-donating methyl group into the picolinate ligand (pic), bis(6- or 3 methylpicolinato)zinc(II) complexes [Zn(6-mpa)2 or Zn(3-mpa)2, respectively] were prepared. The Zn(6-mpa)(2) complex was crystallized as a water adduct [Zn(6 mpa)2(H2O)].H2O, in which two carboxylate oxygens and two pyridine nitrogens of 6 mpa and a water oxygen coordinate to a zinc(II) with a trigonal bipyramidal geometry. By in vitro evaluation of the inhibition of free fatty acid (FFA) release from isolated rat adipocytes in the presence of epinephrine, the insulinomimetic activities of Zn(pic)2, Zn(6-mpa)2, and Zn(3-mpa)2 (IC50=0.64 +/- 0.13, 0.31 +/- 0.05, and 0.40 +/- 0.07 mM, respectively) were found to be higher than those of VOSO(4) (IC50=1.00 mM) and ZnSO(4) (IC50=1.58 +/- 0.05 mM) in terms of IC50 value, the 50% inhibition concentrations for the FFA release from the adipocytes. Then, Zn(6-mpa)2, which exhibited the highest in vitro insulinomimetic activity among three complexes examined, was given at a dose of 3.0 mg (45.9 micromol) Zn/kg body weight to KK-A(y) mice with type 2 diabetes mellitus by daily intraperitoneal injections for 14 days and it was found that the hereditary high blood glucose levels were lowered during the administration of the complex. The improvement of diabetes mellitus was confirmed with the oral glucose tolerance test. PMID- 11862543 TI - Peroxyl adduct radicals formed in the iron/oxygen reconstitution reaction of mutant ribonucleotide reductase R2 proteins from Escherichia coli. AB - Catalytically important free radicals in enzymes are generally formed at highly specific sites, but the specificity is often lost in point mutants where crucial residues have been changed. Among the transient free radicals earlier found in the Y122F mutant of protein R2 in Escherichia coli ribonucleotide reductase after reconstitution with Fe2+ and O2, two were identified as tryptophan radicals. A third radical has an axially symmetric EPR spectrum, and is shown here using 17O exchange and simulations of EPR spectra to be a peroxyl adduct radical. Reconstitution of other mutants of protein R2 (i.e. Y122F/W48Y and Y122F/W107Y) implicates W48 as the origin of the peroxyl adduct. The results indicate that peroxyl radicals form on primary transient radicals on surface residues such as W48, which is accessible to oxygen. However, the specificity of the reaction is not absolute since the single mutant W48Y also gives rise to a peroxyl adduct radical. We used density functional calculations to investigate residue-specific effects on hyperfine coupling constants using models of tryptophan, tyrosine, glycine and cysteine. The results indicate that any peroxyl adduct radical attached to the first three amino acid alpha-carbons gives similar 17O hyperfine coupling constants. Structural arguments and experimental results favor W48 as the major site of peroxyl adducts in the mutant Y122F. Available molecular oxygen can be considered as a spin trap for surface-located protein free radicals. PMID- 11862544 TI - The iron-sulfur center of biotin synthase: site-directed mutants. AB - Biotin synthase contains an essential [4Fe-4S]+ cluster that is thought to provide an electron for the cleavage of S-adenosylmethionine, a cofactor required for biotin formation. The conserved cysteine residues Cys53, Cys57 and Cys60 have been proposed as ligands to the [4Fe-4S] cluster. These residues belong to a C-X3 C-X2-C motif which is also found in pyruvate formate lyase-activating enzyme, lysine 2,3-aminomutase and the anaerobic ribonucleotide reductase-activating component. To investigate the role of the cysteine residues, Cys-->Ala mutants of the eight cysteine residues of Escherichia coli biotin synthase were prepared and assayed for activity. Our results show that six cysteines are important for biotin formation. Only two mutant proteins, C276A and C288A, closely resembled the wild-type protein, indicating that the corresponding cysteines are not involved in iron chelation and biotin formation. The six other mutant proteins, C53A, C57A, C60A, C97A, C128A and C188A, were inactive but capable of assembling a [4Fe-4S] cluster, as shown by Mossbauer spectroscopy. The C53A, C57A and C60A mutant proteins are unique in that their cluster could not undergo reduction to the [4Fe-4S]+ state, as shown by EPR and Mossbauer spectroscopy. On this basis and by analogy with pyruvate formate lyase-activating enzyme and the anaerobic ribonucleotide reductase-activating component, it is suggested that the corresponding cysteines coordinate the cluster even though one cannot fully exclude the possibility that other cysteines play that role as well. Therefore it appears that for activity biotin synthase absolutely requires cysteines that are not involved in iron chelation. PMID- 11862545 TI - The pH-dependent redox inactivation of amicyanin from Paracoccus versutus as studied by rapid protein-film voltammetry. AB - The redox properties of the blue copper protein amicyanin have been studied with slow and fast scan protein-film cyclic voltammetry. At slow scan rates, which reveal the thermodynamics of the redox reactions, the reduction potential of amicyanin depends on pH in a sigmoidal manner, and the data can be analysed in terms of electron transfer being coupled to a single protonatable group with pKa(red)=6.3 and pKa(ox) < or = 3.2 at 22 degrees C. Voltammetry at higher scan rates reveals the kinetics and shows that the low-pH reduced form of amicyanin is not oxidised directly; instead, oxidation occurs only after conversion to the high-pH form. Simulations show that this conversion, which gates the electron transfer, occurs with a rate constant >750 s-1 at 25 degrees C. In order to decrease the rate of the coupled reaction, the experiments were performed at 0 degrees C, at which the rate constant for this conversion was determined to be 35 +/- 20 s-1. Together with evidence from NMR, the results lead to a mechanism involving protonation and dissociation of the copper coordinating histidine-96 in the reduced form. PMID- 11862547 TI - Stopped-flow kinetic study of the peroxidase reactions of mangano-microperoxidase 8. AB - We have investigated the kinetics for the peroxidase-type reaction of mangano microperoxidase 8 (Mn(III)-MP8) by the time-resolved and single-wavelength stopped-flow technique. The formation of intermediate and its subsequent reaction with substrates were studied separately. Oxidation of Mn(III)-MP8 by H2O2 at pH 10.7 yields an intermediate (1) with a rate constant of 2.9 x10(4) M-1 s-1. The formation of 1 exhibits no deuterium solvent isotope effect, favoring the homolytic cleavage of the Mn(III)-MP8 bound hydroperoxide. The rate for the formation of 1 increases sharply as the pH increases and no other intermediate was detected in the entire pH range. Addition of substrate to 1 leads to the regeneration of Mn(III)-MP8. Monitoring the conversion of 1 to Mn(III)-MP8 allows the determination of the substrate reactivity. The substrate reactivity varies by more than two orders of magnitude ranging from 1.04 x 10(6) M-1 s-1 for ascorbic acid to 4.61 x 10(3) M-1s-1 for aniline. It is linearly correlated with the reduction potential for most of the substrates studied, with the easier oxidized species showing greater reactivity. The substrate reactivity drops rapidly as the pH increases. The substrate reactivity at pH 10.7 for the Mn(III)-MP8 system is smaller than that of the corresponding Fe(III)-MP8 system by 2- to 25-fold, depending on the substrate used. PMID- 11862546 TI - The nickel enzyme methyl-coenzyme M reductase from methanogenic archaea: in vitro interconversions among the EPR detectable MCR-red1 and MCR-red2 states. AB - Methyl-coenzyme M reductase (MCR) catalyzes the formation of methane from methyl coenzyme M and coenzyme B in methanogenic archaea. The enzyme contains tightly bound the nickel porphinoid F430. The nickel enzyme has been shown to be active only when its prosthetic group is in the Ni(I) reduced state. In this state MCR exhibits the nickel-based EPR signal red1. We report here for the MCR from Methanothermobacter marburgensis that the EPR spectrum of the active enzyme changed upon addition or removal of coenzyme M, methyl coenzyme M and/or coenzyme B. In the presence of methyl-coenzyme M the red1 signal showed a more resolved 14N-superhyperfine splitting than in the presence of coenzyme M indicating a possible axial ligation of the substrate to the Ni(I). In the presence of methyl coenzyme M and coenzyme B the red1 signal was the same as in the presence of methyl-coenzyme M alone. However, in the presence of coenzyme M and coenzyme B a highly rhombic EPR signal, MCR-red2, was induced, which was found to be light sensitive and appeared to be formed at the expense of the MCR-red1 signal. Upon addition of methyl-coenzyme M, the red2 signal disappeared and the red1 signal increased again. The red2 signal of MCR with 61Ni-labeled cofactor was significantly broadened indicating that the signal is nickel or nickel-ligand based. PMID- 11862548 TI - Comparison of the X-ray absorption properties of the binuclear active site of molluscan and arthropodan hemocyanins. AB - The structural characteristics of oxy- and deoxy-hemocyanins have been investigated using X-ray absorption spectroscopy both in the near-edge (XANES) and for the first shell contribution in the EXAFS region. Several arthropodan and molluscan hemocyanins have been studied in order to trace the inter- and intra phyla differences. The XANES spectra of oxy-hemocyanins of the different species are remarkably similar, consistent with a very strongly conserved co-ordination geometry of the copper active site. In contrast, small but significant differences are observed between the deoxy-forms of arthropodan and molluscan proteins. In particular, the XANES spectra of deoxy-arthropodan hemocyanins (with the exception of L. polyphemus Hc) show a more intense edge feature at approximately 8983 eV. This difference is tentatively assigned to a more planar geometry of the copper-ligands system in the arthropodan rather than in the molluscan proteins. The first shell analysis of the EXAFS modulation is consistent with the presence of n=3Nepsilon(2) imidazole nitrogens at an average distance of 1.92 +/- 0.03 A from copper in all the deoxy-hemocyanins investigated. Binding of dioxygen results for all hemocyanins in the increase of the number of first shell back-scattering atoms to n=5 with average distances of 1.93 A. Alternatively, by separating the contribution of Nepsilon(2) imidazole nitrogens and of peroxide O-atoms, n=3 ligands at 1.98 +/- 0.03 A and n=2 ligands at 1.87 +/- 0.03 A are found. PMID- 11862549 TI - The aminopeptidase from Aeromonas proteolytica can function as an esterase. AB - The aminopeptidase from Aeromonas proteolytica (AAP) can catalyze the hydrolysis of L-leucine ethyl ester ( L-Leu-OEt) with a rate of 96 +/- 5 s-1 and a Km of 700 microM. The observed turnover number for L-Leu-OEt hydrolysis by AAP is similar to that observed for peptide hydrolysis, which is 67 +/- 5 s-1. The k(cat) values for the hydrolysis of L-Leu-OEt and L-leucine- p-nitroanilide ( L- pNA) catalyzed by AAP were determined at different pH values under saturating substrate concentrations. Construction of an Arrhenius plot from the temperature dependence of AAP-catalyzed ester hydrolysis indicates that the rate-limiting step does not change as a function of temperature and is product formation. The activation energy ( Ea) for the activated ES ester complex is 13.7 kJ mol-1, while the enthalpy and entropy of activation at 25 degrees C calculated over the temperature range 298-338 K are 11.2 kJ mol-1 and -175 J K-1 mol-1, respectively. The free energy of activation at 25 degrees C was found to be 63.4 kJ mol-1. The enthalpy of ionization was also measured and was found to be very similar for both peptide and ester substrates, yielding values of 20 kJ mol-1 for L-Leu-OEt and 25 kJ mol-1 for L- pNA. For peptide and L-amino acid ester cleavage reactions catalyzed by AAP, and 6.07, respectively. Proton inventory data suggest that two protons are transferred in the rate-limiting step of ester hydrolysis while only one is transferred in peptide hydrolysis. The combination of these data with the available X-ray crystallographic, kinetic, spectroscopic, and thermodynamic data for AAP provides new insight into the catalytic mechanism of AAP. PMID- 11862550 TI - Characterization of recombinant barley oxalate oxidase expressed by Pichia pastoris. AB - Oxalate oxidase catalyzes the oxidation of oxalate to carbon dioxide and hydrogen peroxide, making it useful for clinical analysis of oxalate in biological fluids. An artificial gene for barley oxalate oxidase has been used to produce functional recombinant enzyme in a Pichia pastoris heterologous expression system, yielding 250 mg of purified oxalate oxidase from 5 L of fermentation medium. The recombinant oxalate oxidase was expressed as a soluble, hexameric 140 kDa glycoprotein containing 0.2 g-atom Mn/monomer with a specific activity of 10 U/mg, similar to the properties reported for enzyme isolated from barley. No superoxide dismutase activity was detected in the recombinant oxalate oxidase. EPR spectra indicate that the majority of the manganese in the protein is present as Mn(II), and are consistent with the six-coordinate metal center reported in the recent X-ray crystal structure for barley oxalate oxidase. The EPR spectra change when bulky anions such as iodide bind, indicating conversion to a five coordinate complex. Addition of oxalate perturbs the EPR spectrum of the Mn(II) sites, providing the first characterization of the substrate complex. The optical absorption spectrum of the concentrated protein contains features associated with a minor six-coordinate Mn(III) species, which disappears on addition of oxalate. EPR spin-trapping experiments indicate that carboxylate free radicals (CO2*-) are transiently produced by the enzyme in the presence of oxalate, most likely during reduction of the Mn(III) sites. These features are incorporated into a turnover mechanism for oxalate oxidase. PMID- 11862551 TI - Kinetics and active site dynamics of Staphylococcus aureus arsenate reductase. AB - Arsenate reductase (ArsC) encoded by Staphylococcus aureus arsenic-resistance plasmid pI258 reduces intracellular arsenate(V) to the more toxic arsenite(III), which is subsequently extruded from the cell. It couples to thioredoxin, thioredoxin reductase and NADPH to be enzymatically active. ArsC is extremely sensitive to oxidative inactivation, has a very dynamic character hampering resonance assignments in NMR and produces peculiar biphasic Michaelis-Menten curves with two V(max) plateaus. In this study, methods to control ArsC oxidation during purification have been optimized. Next, application of Selwyn's test of enzyme inactivation was applied to progress curves and reveals that the addition of tetrahedral oxyanions (50 mM sulfate, phosphate or perchlorate) allows the control of ArsC stability and essentially eliminates the biphasic character of the Michaelis-Menten curves. Finally, 1H-15N HSQC NMR spectroscopy was used to establish that these oxyanions, including the arsenate substrate, exert their stabilizing effect on ArsC through binding with residues located within a C-X5-R sequence motif, characteristic for phosphotyrosine phosphatases. In view of this need for a tetrahedral oxyanion to structure its substrate binding site in its active conformation, a reappraisal of basic kinetic parameters of ArsC was necessary. Under these new conditions and in contrast to previous observations, ArsC has a high substrate specificity, as only arsenate could be reduced ( Km=68 microM, k(cat)/ Km =5.2 x 10(4 )M-1s-1), while its product, arsenite, was identified as a mixed inhibitor ( K*iu=534 microM, K*ic=377 microM). PMID- 11862552 TI - X-ray absorption spectroscopic investigation of Fe(II)-peplomycin and peplomycin derivatives: the effect of axial ligation on Fe-pyrimidine back-bonding. AB - X-ray absorption spectroscopy (XAS) is used to study ferrous complexes of a bleomycin (BLM) congener, peplomycin (PEP), and two of its derivatives, iso peplomycin (ISO) and depyruvamide peplomycin (DP), in which potential axial ligands have been perturbed and removed, respectively. Application of extended X ray absorption fine structure analysis shows an elongation of the short-distance component of the first coordination sphere in DP and ISO relative to PEP. The XAS pre-edge intensity concomitantly decreases with increased axial perturbation. The short-distance component of PEP is correlated to the Fe-pyrimidine bond and is related to the amount of pi-back-bonding. Thus, the XAS analysis of these complexes provides structural information relevant to their differences in O2 reactivity. PMID- 11862553 TI - Nitrite binding to metmyoglobin and methemoglobin in comparison to nitric oxide binding. AB - Nitrite binds reversibly to the ferriheme proteins metmyoglobin and methemoglobin in aqueous buffer solution at a physiological pH of 7.4. The spectral changes recorded for the formation of metMb(NO2-) differ significantly from those observed for the nitrosylation of metMb, which can be accounted for in terms of the different reaction products. Nitric oxide binding to metMb produces a nitrosyl product with Fe(II)-NO+ character, whereas the reaction with nitrite produces an Fe(III)-NO2- complex. The kinetics of the binding and release of nitrite by metMb and metHb were investigated by stopped-flow techniques at ambient and high pressure. The kinetic traces recorded for the reaction of nitrite with metMb exhibit excellent single-exponential fits, whereas nitrite binding to metHb is characterized by double-exponential kinetics which were assigned to the reactions of the alpha- and beta-chains of metHb with NO2-. The rate constants for the binding of nitrite to metMb and metHb were found to be much smaller than those reported for the binding of NO, such that nitrite impurities will not affect the latter reaction. The activation parameters (deltaH++,deltaS(ne),deltaV++) obtained from the temperature and pressure dependence of the reactions support the operation of a dissociative mechanism for the binding and release of nitrite, similar to that found for the binding and release of NO in metMb. PMID- 11862554 TI - Hydrogenases in the "active" state: determination of g-matrix axes and electron spin distribution at the active site by 1H ENDOR spectroscopy. AB - Hydrons and electrons are substrates for the enzyme hydrogenase, but cannot be observed in X-ray crystal structures. High-resolution 1H electron nuclear double resonance (ENDOR) spectroscopy offers a means to detect the distribution of protons and unpaired electrons. ENDOR spectra were recorded from frozen solutions of the nickel-iron hydrogenases of Desulfovibrio gigas and Desulfomicrobium baculatum, in the "active" state ("Ni-C" EPR signal) and analyzed by orientationally selective simulation methods. The experimental spectra were fitted using a structural model of the nickel-iron centre based on crystallographic results, allowing for differences in electron spin distribution as well as the spatial orientation of the g-matrix ( g-tensor), and anisotropic and isotropic hyperfine couplings of the protons nearest to the nickel ion. ENDOR signals, detected after complete deuterium exchange, were assigned to six protons of the cysteines bound to nickel. The assignment took advantage of the substitution of a selenium for a sulfur ligand, which occurs naturally between the [NiFeSe] and [NiFe] hydrogenases from Dm. baculatum and D. gigas, respectively, and was found to affect just two signals. The four signals with the largest hyperfine couplings, including isotropic contributions from 4.5 to 13.5 MHz, were assigned to the beta-methylene protons of the two terminal cysteine ligands, one of which is substituted by seleno-cysteine in [NiFeSe] hydrogenase. The electron spin is delocalized onto the nickel (50%) and its sulfur ligands, with a higher proportion on the terminal than the bridging ligands. The g-matrix was found to align with the active site in such a way that the g1- g2 plane is nearly coplanar (18.3 degrees) with the plane defined by nickel and three sulfur atoms, and the g2 axis deviates by 22.9 degrees from the vector between nickel and iron. Significantly for the reaction of the enzyme, direct evidence for the binding of hydrons at the active site was obtained by the detection of H/D exchangeable ENDOR signals. PMID- 11862555 TI - Nickel-guanine interactions in DNA: crystal structure of nickel-d[CGTGTACACG]2. AB - The aim of this study was to clarify whether Ni2+ ions could bind to guanine bases in a standard B-DNA duplex and eventually induce a B-->Z transition. We have determined by X-ray crystallography at 3.1 A resolution the structure of the alternating deoxynucleotide d(CGTGTACACG), which contains both internal and terminal guanines. The duplex is in the B form. It is shown that nickel ions bind selectively to the N7 atom of guanine 10, which is in an extra-helical position, and guanine 2, which is in the terminal position of the duplex. It does not bind to guanine 4, which lies within a standard B-DNA tract. This simple but unambiguous result proves that nickel ions select between different guanines via steric accessibility. Guanine-Ni2+-guanine bridges among symmetry-related duplexes have also been found. These bridges may explain why Ni2+ ions may act either as a precipitant or a renaturing agent for DNA under certain conditions. The biochemical interaction of nickel with DNA can thus be related to its capacity to specifically bind to B-DNA regions with exposed guanines. Also, from the structural point of view, we have found a terminal cytosine, which forms a C.G:C reverse-Hoogsteen triple structure with a base pair of a neighbor duplex. This type of triplet is seldom found and is here described for the first time for a DNA structure. PMID- 11862556 TI - Studies of the reduction and protonation behavior of tetraheme cytochromes using atomic detail. AB - A comparative study of tetraheme cytochrome c3 molecules from several species was carried out using recently developed theoretical methods based on continuum electrostatics. The binding joint equilibrium of electrons and protons was simulated, revealing the complete thermodynamic aspects of electron-proton coupling in these molecules. The method yields excellent accuracy in terms of midpoint potentials, giving the correct reduction orders in all molecules examined, except for one heme site. The coupling between electrons and protons is shown to be present and significant at physiological pH in all cases. This phenomenon, known as the redox-Bohr effect, though of thermodynamic nature, is shown to have an intrinsic "dynamic" character at the molecular level (in the sense of the empty/occupied fluctuations at the microscopic level), with the binding states of redox and protonatable sites displaying both correlated averages and correlated fluctuations. The protonatable sites more directly involved in the redox-Bohr effect are identified using, among other properties, the statistical correlation between pairs of sites, which automatically reflects indirect effects mediated by other sites. Several sites are identified in this analysis. Propionate D of heme I seems to be the most interesting, generally showing a high correlation not only with its own heme, but also with heme II, corresponding to an indirect stabilization of the reduced forms of both hemes. Other interesting sites are the free histidines of two of the cytochromes and propionate D of heme IV, the latter being potentially associated with redox induced structural changes. Among the set of cytochromes c3 analyzed in this study, significant differences are observed for several properties of the acidic cytochrome included in the set, from Desulfovibrio africanus, supporting the hypothesis of a different functional role. PMID- 11862557 TI - Observation of an isotope-sensitive low-frequency Raman band specific to metmyoglobin. AB - A resonance Raman band involving significantly the iron(III)-histidine stretching (upsilonFe-His) character is identified for metmyoglobin (metMb) through isotope sensitivity of its low-frequency resonance Raman bands, but the identification was not successful for methemoglobin (metHb) and its isolated alpha and beta subunits. A band at 218 cm-1 of natural abundance metMb exhibited a low-frequency shift for 15N-His-labeled metMb (-1.4 cm-1 shift), while the strong porphyrin bands at 248 and 271 cm-1 did not shift significantly. The frequency of the 218 cm-1 band of metMb decreased by 1.6 cm-1 in D2O, probably due to Ndelta deuteration of the proximal His, in a similar manner to that of the upsilonFe-His band of deoxyMb in D2O. This 218-cm-1 band shifted slightly to a lower frequency in H2(18)O, whereas it did little upon 54Fe isotopic substitution (<0.3 cm-1), presumably because of the six-coordinate structure. The lack of the 54Fe-isotope shift shows that the 218-cm-1 band is specific to metMb and not due to the deoxy species. The intensity of this band decreased for hydroxymetMb and was indiscernible for cyanometMb. For metHb and its alpha and beta subunits, however, the frequencies of the band around 220 cm-1 were not D2O sensitive. These results suggest an assignment of the band around 220 cm-1 to a pyrrole tilting mode, which significantly contains the Fe-His stretching character for metMb but scarcely for metHb and its subunits. The differences in the isotope sensitivity of this band in different proteins are considered to reflect the heme distortion from the planarity and the Fe-His geometry specific to individual proteins. PMID- 11862560 TI - Sources of clinical referrals to an urban coloproctology unit in Italy. AB - Prompt and appropriate referrals to colorectal surgeons result in better clinical and more cost-effective outcome: the question that then arises is how patients with large bowel diseases get in contact with the specialist. The aim of the present research was to investigate the sources of clinical referrals of 1000 patients attending a dedicated coloproctology unit. One thousand consecutive new patients attending the private Coloproctology Unit of Rome were prospectively evaluated from May 1995 through December 1999. For each patient, the following data were collected: age, gender, source of referral, and disease classified as benign anal diseases, neoplasms, functional disorders or inflammatory bowel disease (IBD). There were 569 patients with benign anal disease, 334 with functional disorders, 57 with neoplasms, and 40 with IBD. Sources of referral were: surgeons (32.6%), previous patients (23.6%), other specialists (22.8%), general practitioners (11.8) and others (9.2%). Overall, referrals from non medical sources were 32.8%, whereas 67.2% of the cases were referred by other colleagues. Most of the referring specialists were surgeons or gastroenterologists, who sent 304 patients, whereas 9.4% of the cases were referred by other colorectal surgeons. Previous patients who were satisfactorily cured sent 23.6% of the cases. Only 1.1% of patients were referred by health insurance companies and 0.2% found the Unit through Internet. Colleagues who referred patients to the coloproctologist sent mainly cases with benign anal diseases and functional disorders. Few patients were referred for colorectal cancer and IBD as these diseases are routinely treated by general surgeons and their management is expensive in a private hospital for patients without insurance coverage. In conclusion, GPs, media, health insurance and Internet may be the most valuable targets of an information campaign, as their role as sources of referral was lower than expected. PMID- 11862561 TI - Tumor size is irrelevant in predicting malignant potential of carcinoid tumors of the rectum. AB - The malignant potential and prognosis of rectal carcinoids are said to be related to tumor size. Our study assessed if size could predict the malignant potential and hence its management. All patients in the Department of Colorectal Surgery, Singapore General Hospital, who underwent surgery for rectal carcinoid tumors between February 1991 and September 2000 were analyzed. Twenty patients (11 men), median age 48 years (range, 33-77 years) were studied. Median follow-up was 40 months (range, 5-120 months). The median tumor diameter was 2.5 cm (range, 0.1 5.0 cm). Eleven patients underwent radical resection and 9 patients had local resection for a presumed benign tumor. Morbidity was 15% and postoperative death was 5%. Overall median survival was 24 months (range, 5-120 months). One patient had an anterior resection for rectal adenocarcinoma but had an incidental 0.1-cm carcinoid tumor near the resection margin which on histology was found to have carcinoid tumor metastasis to 2 out of 12 lymph nodes. In conclusion, tumor size cannot predict malignant potential as even small tumors (<1 cm) can be malignant. Accurate preoperative staging with radical surgery may be required. PMID- 11862562 TI - Total colectomy with ileorectal anastomosis leads to appreciable loss in quality of life irrespective of primary diagnosis. AB - Total colectomy with ileorectal anastomosis (TC) is a well-accepted procedure for many colonic pathologies but data on faecal incontinence and related quality of life after TC are lacking. The aims of this study were to assess the long-term bowel frequency, degree of incontinence and quality of life with respect to faecal incontinence and to compare them with the outcome for TC for different diagnostic groups. We identified 54 patients who had undergone TC at Singapore General Hospital and interviewed them using two questionnaires: the faecal incontinence quality of life (FIQL) scale and the Wexner faecal incontinence score (WS). The patients were allocated in 3 groups based on the primary diagnosis leading to operation, i. e. slow-transit constipation or megacolon (STC), colonic neoplasm (CA) and complicated pan-colonic diverticular disease (DD). Median bowel frequencies for STC and DD groups were 2.5/day; for CA, it was 3.5/day (p=0.042). There was no significant difference in the FIQL score and WS between the groups. Eleven patients had some degree of faecal incontinence based on WS. Many patients (20.4%) with perfect continence had fear of faecal leakage affecting their quality of life. In conclusion, patients with frequent stools do not need to have incontinence to suffer from the fear of it. The primary pathology leading to TC made no difference to the faecal incontinence or bowel urgency problems. PMID- 11862563 TI - Parastomal hernia surgery: personal experience with use of polypropylene mesh. AB - Following stoma construction, parastomal hernia is the most frequent complication. Many surgical techniques have been postulated for these patients, and prosthetic surgery represents the first-choice treatment. We report our personal experience with 8 cases of parastomal hernia in patients submitted to abdominal perineal resection according to Miles, for carcinoma of the lower rectum. Polypropylene mesh was shaped according to the size of the fascial defects, characterized by a romboid incision about 4 cm in length. The mesh was placed in suprafascial position, after suturing the fascial tear. One case of wound infection occurred and, to date, none of the patients have presented with recurrence after a 3-year follow-up. In conclusion, the use of polypropylene mesh for parastomal hernia repair represents a safe and successful technique. PMID- 11862564 TI - A prospective randomised study of drains in infra-peritoneal rectal anastomoses. AB - Although increasing evidence suggests that prophylactic drainage after intra peritoneal colorectal anastomoses is unnecessary, drains for infra-peritoneal rectal anastomoses, where the leak rate is higher, are widely employed still. The aim of this study was to assess the effect of prophylactic drainage after anastomosis below the peritoneal reflection. All patients attending one specialist unit over an 8-month period for elective rectal cancer resection with an infra-peritoneal anastomosis were randomised to drainage or no drainage. The incidence of anastomotic leak and complications specific to the drain as well as other complications were compared. Fifty-nine patients were analysed (31 with drain). Twenty-five of the drained and 16 of the no-drain patients had a defunctioning stoma (p=ns). The groups were comparable for demographic data, operation and anastomotic height from the anal verge. There were three leaks (10%) in the drain group and five leaks (18%) in the no-drain group (p=ns). There were 2 (7%) patients in each group with a clinical leak. There were no specific drain complications and the incidence of other complications was similar in both groups. In conclusion, this study supports the contention that there is no difference in morbidity with or without the use of a drain for infra-peritoneal anastomoses. PMID- 11862565 TI - Whitehead's hemorrhoidectomy. A useful surgical procedure in selected cases. AB - At the present time Milligan-Morgan's operation is the most diffusely employed and is widely considered to be the most effective of the various surgical techniques for the treatment of hemorrhoids. In this study we report our experience with Whitehead's radical hemorrhoidectomy. In a 5-year period, 1450 patients with hemorrhoids were treated at our Coloproctologic Unit. We routinely carry out the Milligan-Morgan operation. Nevertheless, in 26 patients the Milligan-Morgan operation was judged to be impossible to perform, in that the prolapsed hemorrhoids were completely irreducible and it was not possible to distinguish and separate the three piles. These patients thus underwent Whitehead's radical hemorroidectomy. All the patients who underwent Whitehead's operation were discharged within the fifth post-operative day. No episodes of incontinence were observed in any patient. The patients were followed for three years after the operation. In only one case did we verify an anal substenosis three months after the operation, which resolved after the use of anal dilators for one month. The stenosis did not recur in the course of follow-up. There were no cases of mucosal ectropion. In conclusion, the type of hemorrhoidectomy which a surgeon performs is primarily based on the surgeon's experience and training. Nevertheless, a competently performed Whitehead's hemorrhoidectomy can give satisfying results. These results are explained by improved knowledge of the anatomy of the anal region and a more accurate surgical technique. On the basis of our experience we believe that Whitehead's hemorrhoidectomy still has its place in selected cases with precise indications. PMID- 11862566 TI - Abdominoperineal resection for locally recurrent rectal cancer. AB - To evaluate whether surgical resection confers survival advantages in selected patients with resectable locally recurrent rectal cancer, data on 430 patients who underwent R0 resection for primary rectal cancer were prospectively collected over a 14-year period. Resection of recurrent disease was considered R0 when all cancer tissue was resected with microscopically tumor-free surgical margins. Microscopic evidence of disease at resection margins was considered an R1 resection. Recurrent disease was detected in 158 of 430 patients. Local recurrence was found in 91 patients, including (79%) with resection-site relapse only. These patients were considered for surgery unless defined unresectability criteria were met. A total of 35 patients who had abdominoperineal excision following anterior resection were studied retrospectively. Mortality associated with the procedure was 3% and morbidity was 20%. The resection was R0 in 12 patients, while microscopic margins were involved in 23 patients. 10 patients had extended resection of one or several adjacent organs Median operating time and blood loss were 250 min and 500 ml respectively. Median hospital stay was 25 days. 21/23 R1 patients received postoperative radiotherapy. Return to normal activity occurred at 8.2 (SD 4.2) weeks. No patients were lost to follow-up. Overall median survival was 26.4 months; 5-year survival was 25.4%. In spite of several survival predictors at univariate analysis, R0 or R1 resection was the only independent predictor of survival at multivariate analysis (add ratio 112.7, 95% CI 3.6-3500, p=0.007). Median survival rate was not reached at the 146-month follow-up in patients with R0 resection. Median survival rate was 16.6 months in patients with R1 resection. In conclusion, uninvolved microscopic margins produce long-term survivors after surgical resection for locally recurrent rectal cancer. PMID- 11862567 TI - Dynamic imaging of pelvic floor with transperineal sonography. AB - Real-time transperineal sonography has enhanced the appreciation of morphology and dynamics of the pelvic floor. Standard images are obtained from longitudinal and axial planes by placing the transducer between the vagina and rectum. This fast, effective, noninvasive and inexpensive examination represents the preferred initial diagnostic imaging tool for women with pelvic floor dysfunctions, such as prolapse and incontinence. PMID- 11862568 TI - Perineal endometriosis in episiotomy scar with anal sphincter involvement. AB - Perineal endometriosis, especially with anal sphincter invasion, is a rare occurrence. We present a patient with perineal endometriosis in an episiotomy scar with anal sphincter involvement. The endometriotic mass was completely excised under general anesthesia with portions of the episiotomy scar and external anal sphincter. The procedure was followed by overlapping sphincter reconstruction. The excised mass was sent for microscopic examination, which confirmed endometriosis. The postoperative course was without complications. One year after the operation, the woman is asymptomatic and fully continent. Complete excision including a part of the anal sphincter with primary sphincteroplasty is the best treatment for perineal endometriosis involving the anal sphincter. PMID- 11862569 TI - Life-threatening urinary retention after haemorrhoidectomy and internal sphincterotomy. AB - Postoperative urinary retention (UR) is a common complication following haemorrhoidectomy. This report presents a female patient who developed long lasting UR after haemorrhoidectomy and internal sphincterotomy, which progressed to renal failure. The precipitating cause was represented by a non-healing anal ulcer and excruciating anal pain at evacuation. An unhealed anal wound, an inappropriate low-fibre diet, and excruciating anal pain commonly represent the key factors initiating the sequence ending into UR. History, presentation, diagnostic work-up, and treatment for this patient are described. PMID- 11862572 TI - Experimental model of anorectal transplantation. PMID- 11862576 TI - Heart Failure Awareness 2002--and beyond. PMID- 11862573 TI - Combined abdominoperineal repair of enterocele and rectocele. PMID- 11862578 TI - Impaired endothelium-mediated vasodilation in heart failure: clinical evidence and the potential for therapy. AB - Numerous studies in the last decade have clearly shown an attenuated endothelium dependent vasodilation in patients with chronic heart failure. This abnormality has been demonstrated in the peripheral, pulmonary, and coronary circulation in patients with both ischemic and nonischemic cardiomyopathy; its magnitude correlates with the severity of symptoms. Endothelial dysfunction in patients with cardiomyopathy and a relatively new onset of symptoms suggests that change in endothelial function occurs early in the course of the disease. In contrast to other circulatory beds, renal circulation has shown significant vasodilatory response to endothelial stimulation. The development of endothelial dysfunction may not be homogeneous, and its magnitude may differ among circulatory systems. Although the clinical implications of the attenuated endothelium-dependent vasodilation in heart failure are not clear, this condition may lead to decreased organ perfusion, impaired exercise tolerance, and progression of disease. Many therapeutic interventions have resulted in improvement of endothelial function in patients with heart failure. Some of these interventions have also proven effective in enhancing exercise capacity, symptoms, and survival in patients with heart failure. This association suggests a therapeutic role for improvement of endothelial function in patients with chronic heart failure. PMID- 11862577 TI - Effects of metoprolol CR/XL on mortality and hospitalizations in patients with heart failure and history of hypertension. AB - BACKGROUND: We describe the effect of controlled-release/extended-release (CR/XL) metoprolol succinate once daily on mortality and hospitalizations among patients with a history of hypertension complicated by chronic systolic heart failure. METHODS AND RESULTS: We enrolled 3,991 patients with chronic heart failure of New York Heart Association functional class II-IV with an ejection fraction of < or = 0.40, stabilized with optimum standard therapy, in a double-blind randomized placebo-controlled study. A total of 1,747 patients (44%) had a history of hypertension; 871 were randomized to receive metoprolol CR/XL and 876 to receive placebo. Treatment with metoprolol CR/XL compared with placebo resulted in a significant reduction in total mortality (relative risk [RR], 0.61; 95% confidence interval [CI], 0.44-0.84; P =.0022), mainly because of reductions in sudden death (RR, 0.51; 95% CI, 0.33-0.79; P =.0022) and mortality from worsening heart failure (RR, 0.49; 95% CI, 0.25-0.99; P =.042). Total number of hospitalizations for worsening heart failure was reduced by 30% in the metoprolol CR/XL group compared with placebo (P =.015). Metoprolol CR/XL was well tolerated: 12% fewer patients withdrew from study medication (all-cause) compared with placebo (P =.048). CONCLUSIONS: A subgroup analysis of MERIT-HF shows that patients with heart failure and a history of hypertension received a similar benefit from metoprolol CR/XL treatment as all patients included in the total study. PMID- 11862579 TI - Circulating levels of interleukin 18 reflect etiologies of heart failure: Th1/Th2 cytokine imbalance exaggerates the pathophysiology of advanced heart failure. AB - BACKGROUND: Proinflammatory cytokines such as tumor necrosis factor alpha play an important role in the pathophysiology of CHF. However, the mechanisms of immune activation in CHF remain unknown. Interleukin (IL)-18, a newly discovered cytokine with pleiotropic activities, is known to induce proinflammatory cytokines, chemokines, nitric oxide, and prostaglandins. METHODS AND RESULTS: We studied 86 patients with New York Heart Association functional class II-IV heart failure. Mean age was 62 years, 20 were women, and mean left ventricular ejection fraction was 34.8%. Circulating levels of IL-18 and IL-10, high-sensitivity testing for C-reactive protein, and brain natriuretic peptide levels were determined. Serum IL-18 concentrations were significantly higher in patients with NYHA class IV than in patients with classes II and III (P <.001). The serum level of IL-18 and the ratio of IL-18 to IL-10 were greater in patients with ischemic cardiomyopathy than in those with dilated cardiomyopathy. CONCLUSIONS: Th1/Th2 cytokine imbalance exists in patients with advanced CHF according to various etiologies of CHF. The findings suggest an important role for IL-18 in the pathophysiology of CHF and provide a direction for more specific immunomodulating therapy. PMID- 11862580 TI - Cardiac troponin T lysine 210 deletion in a family with dilated cardiomyopathy. AB - BACKGROUND: The gene for cardiac troponin T (TNNT2) is 1 of 7 autosomal disease genes implicated in familial dilated cardiomyopathy (FDC). Identical deletions in exon 13 of TNNT2 have been reported in 2 families with FDC, but little is known about the frequency of this deletion among patients with FDC and idiopathic dilated cardiomyopathy (IDC) and the associated phenotype. METHODS AND RESULTS: Exon 13 of the cardiac troponin T gene was sequenced in 61 subjects with FDC and 53 subjects with IDC. A 3-base pair deletion (DeltaLys210), identified in 1 family with at least 7 clinically affected family members, is reported. Age of disease onset and disease severity varied widely among affected individuals; phenotypic findings included dilated cardiomyopathy, sudden cardiac death, conduction system disease including atrial fibrillation and atrioventricular block, and heart failure. Sudden-onset, rapidly progressive disease was observed in younger individuals. CONCLUSIONS: Cardiac troponin T exon 13 lysine deletions can cause FDC of varying severity and are an important but uncommon cause of FDC. PMID- 11862581 TI - A sodium channel enhancer, LY341311, increases myocardial contractile performance without increasing heart rate in conscious normal dogs: a comparison with dobutamine. AB - BACKGROUND: Catecholamines and many inotropic agents increase cardiac contractility but also cause excessive myocardial O2 consumption (MVO2). We determined if the novel Na+ channel enhancer LY341311, which increases myocardial contractility independent of beta receptors, can produce significant cardiac inotropic effects compared with dobutamine but at lower oxygen cost in conscious dogs. METHODS AND RESULTS: Mongrel dogs were chronically instrumented for measurement of arterial pressure, left ventricular (LV) pressure and internal diameter, coronary blood flow, and arterial and coronary sinus O2 content. Both LY341311 and dobutamine produced dose-dependent increases in LV dP/dt, dP/dt/40, fractional shortening, and cardiac stroke work and minute work estimated from the LV pressure-diameter loop. The major difference between LY341311 and dobutamine was an opposing effect on heart rate with LY341311 slightly reducing it but dobutamine markedly increasing it. LY341311 caused a significantly smaller increase in MVO2 than dobutamine (P <.05) and produced similar cardiac inotropic effects, yielding a higher cardiac mechanical efficiency than dobutamine. However, after pacing to match heart rate with dobutamine LY341311 increased MVO2 markedly, approaching the same level as with dobutamine. CONCLUSIONS: The novel Na+ channel enhancer LY341311 caused significant increases in myocardial contractility and contractile performance without increasing heart rate. It had a beneficial energetic effect on the heart with significantly less O2 cost and improved cardiac mechanical efficiency. PMID- 11862582 TI - Temporal regression of myocyte hypertrophy in hypertensive, heart failure-prone rats treated with an AT1-receptor antagonist. AB - BACKGROUND: A recent study showed reverse remodeling of left ventricular myocyte shape when the type 1 angiotensin II (AT1)-receptor antagonist L-158,809 was administered to spontaneously hypertensive heart failure (SHHF) rats 4 months before the onset of failure. The aim of this study was to characterize temporally early treatment-induced reverse remodeling at the organ and cellular level by echocardiography and morphometry of isolated left ventricular myocytes. METHODS AND RESULTS: L-158,809 was administered to 9-month-old SHHF rats. Blood pressure normalized shortly after initiation of treatment. Isolated myocytes were collected in terminal experiments to assess cell remodeling. L-158,809 reduced myocyte volume and cross-sectional area significantly after 1 week of treatment with maximal regression of hypertrophy, including reduced cell length, obtained by 4 weeks. Reduced wall thickness was clearly detectable by echocardiography within 4 weeks after initiation of treatment. CONCLUSIONS: Regression of left ventricular myocyte hypertrophy occurred rapidly after initiation of AT1-blocker therapy in SHHF rats and was completed within 1 month. Regression of myocyte hypertrophy was associated with reduced wall thickness, which was detected consistently by echocardiography within 1 month after initiation of treatment. PMID- 11862583 TI - Anticoagulation in patients with dilated cardiomyopathy and sinus rhythm: a critical literature review. AB - BACKGROUND: The risk of thromboembolism in chronic heart failure and the risk-to benefit ratio of anticoagulation in this population are poorly defined. METHODS AND RESULTS: A PubMed/MEDLINE search of published trials was performed. Twenty four studies were identified after exclusion of individual case reports. All studies were prospective or retrospective observational reports, either independent studies or secondary analyses of prospective clinical trials in patients with heart failure. Prevalence estimates ranged of thromboemboli ranged from 3% to 50% and incidence estimates ranged from 1.5 to 3.5/100 patient-years. Although no randomized data of therapeutic anticoagulation were identified, a secondary analysis of one study suggested event reduction in patients receiving warfarin anticoagulation; other studies failed to suggest such benefit. Overall bleeding estimates in warfarin-treated patients ranged from 2.3 to 6.8/100 patient-years. Intracranial hemorrhage rates were 0.62 to 0.9/100 patient-years but increased with age. Only one study suggested that aspirin was beneficial in reducing clinically significant emboli. CONCLUSIONS: Although patients with chronic heart failure and left ventricular dilation have a higher risk of thromboembolism, data are insufficient to recommend warfarin or aspirin prophylaxis in the absence of additional indications for such therapy. PMID- 11862584 TI - Pathophysiology of renal disease associated with liver disorders: implications for liver transplantation. Part I. AB - Renal and hepatic function are often intertwined through both the existence of associated primary organ diseases and hemodynamic interrelationships. This connection occasionally results in the chronic failure of both organs, necessitating combined liver-kidney transplantation (LKT). Since 1988, more than 850 patients in the United States have received such transplants, with patient survival somewhat less than that for patients receiving either organ alone. Patients with renal failure caused by acute injury or hepatorenal syndrome have classically not been included as candidates for combined transplantation because of the reversibility of renal dysfunction after liver transplantation. However, the rate and duration of renal failure before liver transplantation is increasing in association with prolonged waiting list times. Thus, the issue of acquired permanent renal damage in the setting of hepatic failure continues to confront the transplant community. The following article and its sequel (Part II, to be published in vol 8, no 3 of this journal) attempt to review the problem of primary and secondary renal disease in patients with end-stage liver disease, elements involved in renal disease progression and recovery, the impact of renal disease on liver transplant outcome, and results of combined LKT; outline the steps in the pretransplantation renal evaluation; and provide the beginnings of an algorithm for making the decision for combined LKT. PMID- 11862585 TI - Donor morbidity associated with right lobectomy for living donor liver transplantation to adult recipients: a systematic review. AB - The aim if this study is to determine donor morbidity associated with right lobectomy for living donor liver transplantation (LDLT) to adult recipients through a systematic review of the published literature. Data sources were English-language reports on donor outcome after LDLT. MEDLINE (1995 to June 2001) was searched using the MeSH terms "living donors" and "liver transplantation." Limits were set for human only and English language only. Bibliographies of retrieved references were cross-checked to identify additional reports; 211 reports were obtained. Population studies and consecutive and nonconsecutive series were included. All studies reported at least one of the following outcomes specific to living donors (LDs) of right hepatic lobes to adult recipients: surgical and hospital complications, length of hospital stay, readmissions, recovery time, return to predonation occupation, health-related quality of life, or mortality. Abstracts of relevant articles were reviewed independently using predetermined criteria, and appropriate articles were retrieved. Study design and results were summarized in evidence tables. Summary statistics of combined data were performed when possible. Twelve studies met the inclusion criteria. Data on donor morbidity associated with right lobectomy are limited. On the basis of reported data, morbidity associated with LD right lobectomy ranges from 0% to 67%. In conclusion, reported morbidity associated with right lobe donation for LDLT varies widely. Standardized definitions of morbidity and better methods for observing and measuring outcomes are necessary to understand and potentially improve morbidity. Future studies assessing LD outcomes should report donor outcome more explicitly. PMID- 11862586 TI - Donor safety in living related liver transplantation: underestimation of the risks for deep vein thrombosis and pulmonary embolism. AB - Optimal safety for donors is a necessary condition for living related liver transplantation to expand. Although the risks for complications directly related to surgical intervention have been carefully evaluated, the extent and nature of other complications, such as pulmonary embolism, associated with living donation have not been clearly anticipated. We report a case of severe pulmonary embolism followed by ulcer-related upper digestive tract bleeding in an adult donor after right hepatectomy. In this donor, we found an unexpected predisposing coagulation disorder (increased von Willebrand factor activity) postoperatively. This finding led us to include systematic screening for coagulation disorders during evaluation of donors, a policy which led us thereafter to reject definitely or temporarily some candidates for donation. Although the cost-effectiveness of such investigations is not definitely established, we strongly recommend a specific approach to improve donor safety. In addition, we emphasize that the major complications observed in living donors should be systematically reported in an international database. PMID- 11862587 TI - How safe are donors in adult-to-adult living related liver transplantation? PMID- 11862588 TI - Immunoprophylaxis with basiliximab, a chimeric anti-interleukin-2 receptor monoclonal antibody, in combination with azathioprine-containing triple therapy in liver transplant recipients. AB - Acute graft rejection remains a major problem among additional sequelae in liver transplant recipients. Basiliximab, a chimeric monoclonal antibody with high affinity for the CD25 chain of the interleukin-2 receptor, has significantly reduced the incidence of acute rejection episodes in renal transplant recipients. This single-arm, open-label, multicenter study investigated the efficacy and tolerability of basiliximab immunoprophylaxis in adult patients undergoing first elective liver transplantation. One hundred one patients (70 hepatitis C virus [HCV]-negative patients, 31 HCV-positive patients) were administered basiliximab, 20 mg, by intravenous bolus injection the day of transplantation (day 0) and day 4. In addition, all patients were administered triple immunosuppressive therapy with cyclosporine, steroids, and azathioprine. The efficacy of basiliximab was assessed by conventional parameters, and tolerability was assessed by the incidence of adverse events, infections, and laboratory test result abnormalities. At 6 months, the incidence of first acute biopsy-confirmed rejection episodes was 22.8%. Rejections were more frequent in the HCV-positive (29.0%) than HCV-negative subgroup (20.0%; P =.441). No rejection episode was graded histologically as severe, and no patient required antibody therapy for the management of acute rejection. Ten patients (9.9%) required treatment with tacrolimus for acute rejection episodes. Patient and graft survival rates at 12 months were 90.1% and 88.1%, respectively. Basiliximab caused no injection-site reactions, anaphylaxis, or cytokine release syndrome. Five malignancies were reported at 12 months: of these, three malignancies predated transplantation surgery. Compared with earlier studies, the addition of basiliximab immunoprophylaxis to triple immunosuppressive therapy provides increased efficacy in reducing the incidence of acute rejection episodes, with no clinically significant increase in adverse events. PMID- 11862589 TI - Improved treatment response with basiliximab immunoprophylaxis after liver transplantation: results from a double-blind randomized placebo-controlled trial. AB - Basiliximab, a high-affinity chimeric monoclonal antibody, is effective in reducing acute rejection episodes in renal allograft recipients. We assessed the ability of this antibody to similarly improve the outcome in liver transplant recipients. Adult recipients of a primary cadaveric liver transplant were randomized to treatment, stratified by hepatitis C virus (HCV) seropositivity. Patients were administered 40 mg of basiliximab (n = 188) or placebo (n = 193) as two 20-mg bolus injections days 0 and 4, plus cyclosporine and steroids. Primary efficacy variables were biopsy-confirmed acute rejection and its composite end point, including death or graft loss, and were assessed at 6 and 12 months and by HCV cohort. Because of differential efficacy responses between HCV-positive and HCV-negative cohorts, an additional analysis incorporating HCV recurrence as a component of treatment failure, termed problem-free transplant, was introduced. Safety and tolerability were monitored over the 12 months of the study. All 381 patients were assessable, and no meaningful differences in background characteristics were apparent between treatment groups. Biopsy-confirmed acute rejection rates 6 months after transplantation were 35.1% in the basiliximab group versus 43.5% in the placebo group. For death, graft loss, or first biopsy confirmed acute rejection, rates were 44.1% versus 52.8%, respectively. The reduction in rejection episodes was concentrated in the HCV-negative cohort (14.5% relative to placebo; P =.034), with a much smaller difference (2.9%) in the HCV-positive cohort. For HCV-positive patients, problem-free transplant was shown at 12 months in 26.6% of the basiliximab group versus 11.6% in the placebo group (P =.020) and for all patients at 12 months in 39.7% of the basiliximab group versus 30.1% in the placebo group (P =.035). The incidence of infection and other adverse events was similar across the two treatment groups. There were 56 deaths (25 deaths, basiliximab group; 31 deaths, placebo group) over the 12-month study. The intravenous bolus injection was well tolerated. Immunoprophylaxis with 40 mg of basiliximab, in combination with cyclosporine and steroids, reduces the incidence of acute rejection episodes with no clinically relevant safety or tolerability concerns. The influence of HCV recurrence on efficacy results can be accounted for in future trials by using the concept of problem-free transplant, incorporating recurrence as a component of treatment failure. PMID- 11862590 TI - Pros and cons of using interleukin-2 receptor antibodies in liver transplant recipients. PMID- 11862591 TI - Effects of portal vein arterialization on liver regeneration after partial hepatectomy in the rat. AB - Although portal venous supply is considered essential to preserve hepatic integrity, in this study, effects of portal arterialization on liver regeneration were evaluated in a rat model of partial hepatectomy (PH). Ninety-six Lewis rats were randomly assigned to four groups of 24 rats each: PH only (group 1), PH with either venous or arterialized portal supply (groups 2 and 3, respectively), and PH without portal supply (group 4). Liver regeneration rate (LRR), 5-bromo-2 deoxyuridine (BrdU) labeling index, and liver biological characteristics were assessed on days 1, 2, 3, and 7. Compared with group 1, all tested rats had a marked body weight loss after surgery, and only rats in group 4 showed no signs of recovery on day 7. With maintained portal inflow (groups 1, 2, and 3), LRRs increased steadily to day-7 values of 89.2% +/- 11.8%, 81.4% +/- 8%, and 77.4% +/ 9.4%, respectively (P = not significant), and 24-hour peak values of BrdU labeling index were 159 +/- 26, 157 +/- 42, and 149 +/- 48, respectively (P = not significant). Conversely, rats deprived of portal supply (group 4) showed profound inhibition of these two parameters (14 +/- 13; P <.01;32.1% +/- 7.7%; P <.001, respectively). These results indicate that proper portal blood supply is essential to initiate and maintain liver regeneration after PH. With an equivalent portal inflow rate of either venous or arterial source, the hepatic regeneration response can be sustained. PMID- 11862592 TI - Aberrations in hemostasis and coagulation in untreated discordant hepatic xenotransplantation: studies in the dog-to-pig model. AB - Discordant liver xenotransplantation is a poorly explored entity. Data from the few large animal studies of hepatic xenotransplantation suggest that severe hemorrhage is encountered. The purpose of the studies described here is to characterize the nature of the hemorrhage that accompanies liver xenotransplantation. Canine livers were transplanted into porcine recipients, and lethal hemorrhage was encountered. Analysis of recipient blood showed that factors V, IX, and X were present in adequate levels until after the hemorrhage appeared, suggesting that coagulation factor loss was the result and not the cause of hemorrhage. Platelet counts decreased dramatically in recipients within minutes of graft reperfusion. There also was no evidence of clotting activity in the blood of recipients of liver xenografts within minutes of graft reperfusion. This loss of clotting activity was specific to liver xenografts, was not seen in renal xenografts with or without venovenous bypass, and also was absent in pig-to pig liver allografts. In brief, the hemorrhage that accompanies liver xenotransplantation occurs because of a decrease in the number and function of circulating platelets in the recipient. PMID- 11862593 TI - Correlation between intraoperative blood flows and hepatic artery strictures in liver transplantation. AB - Hepatic artery strictures (HASs) may be a source of morbidity and mortality in liver transplant recipients. This study evaluated the potential correlation between intraoperative arterial and venous blood flows measured after implantation of the liver allograft and the occurrence of postoperative HASs requiring repair. Prospectively collected data from 1,038 patients with complete data sets who underwent initial orthotopic liver transplantations between December 1984 and December 1999 were used. Electromagnetic flow measurements were routinely obtained in these cases. Hepatic artery and portal vein patency were reassessed routinely according to our protocol in the first postoperative day by Doppler ultrasound. When considered hemodynamically significant, strictures were corrected. There was a 6.2% incidence (n = 64) of hepatic artery stenosis in our transplant population. When considered as a whole, the hepatic artery stenosis group had lower intraoperative flow volumes than transplant recipients who did not develop strictures (mean flows, 452 v 518 mL/min, respectively; P =.025). The hepatic artery stenosis group also had lower intraoperative portal vein flows compared with the group without hepatic artery stenosis (1.80 v 2.11 L/min, respectively; P =.0043). Strictures were less frequent among transplant recipients with cryptogenic cirrhosis. We did not observe differences among the groups for retransplantation or patient and graft survival. In our series, there was a 6.2% incidence of postoperative HASs. We observed a significant association between intraoperative hepatic artery and portal vein flows and postoperative HASs. PMID- 11862594 TI - Rapid improvement of autonomic and peripheral neuropathy after liver transplantation: a single case report. AB - Peripheral and autonomic neuropathies are known but often unrecognized associations of cirrhosis from any cause. The pathogenesis of these effects are ill understood. Liver transplantation has been shown to reverse autonomic manifestations, but little evidence exists for an effect on peripheral neuropathy. This case report documents improvement in peripheral and autonomic neuropathy in a 40-year-old man with hepatitis B virus--related cirrhosis. A return to normal neurophysiological function was seen within 9 months of successful liver transplantation, suggesting a metabolic, rather than a structural, cause of such changes in the peripheral nervous system. PMID- 11862595 TI - Outflow reconstruction in recipients of right liver graft from living donors. AB - Right-lobe transplantation is now a commonly used procedure in living donor liver transplantation (LDLT) to adult recipients. However, the risk for outflow obstruction is still an issue in LDLT. The right hepatic vein (RHV) was anastomosed end to end to the graft hepatic vein without unfavorable tension on the anastomosis. The anterior wall of the recipient hepatic vein was incised longitudinally, and a V-shaped vein graft was patched to form a wide and long orifice. This new hepatic venoplasty was used in 14 adult patients who received right liver grafts and gave good results without stenosis of the hepatic venous anastomosis or other complications. Our new technique may be useful in recipients of a right liver graft when the recipient or graft RHV is not long enough. PMID- 11862596 TI - Successful pregnancy after porcine bioartificial liver treatment and liver transplantation for fulminant hepatic failure. PMID- 11862597 TI - Obesity and the liver transplant recipient. PMID- 11862599 TI - Letter from the editors. PMID- 11862600 TI - ADA targets oral cancer awareness. PMID- 11862598 TI - Living donor liver transplantation: summary of a conference at The National Institutes of Health. AB - Living donor liver transplantation for adults was developed only recently in an attempt to increase the pool of donor organs; to reduce morbidity and mortality; and to improve the long-term survival of patients in need of liver transplant. Within a few brief years, this procedure has gained widespread support by both the public and transplant community. The procedure will soon be performed by nearly 80% of all liver transplant programs in the United States. Unfortunately, the long-term risks of the procedure to the recipient and especially the donor remain undefined. In response to the rapid growth and enthusiasm for this procedure, the National Institutes of Health sponsored a workshop, the goals of which were to review the scientific, medical, and nonmedical issues associated with living donor liver transplantation, and to define questions for future basic and clinical investigations which could improve the success and applicability of this procedure. PMID- 11862601 TI - Beyond our wildest dreams: one answer to access to care. PMID- 11862602 TI - Whatever works: (way) behind the scenes at the Minnesota Dental Association's Star of the North Meeting. PMID- 11862603 TI - Just look over your shoulder: the mentor process in dentistry. PMID- 11862604 TI - Managed care in the dental marketplace: 2000. PMID- 11862605 TI - The patient-centered practice. PMID- 11862606 TI - Modulation by divalent cations of GABA rho 1 receptor from human retina expressed in Xenopus oocytes. AB - OBJECTIVE: To investigate functional homooligomeric GABA rho 1 receptors expressed in Xenopus oocytes and the modulation of divalent cations. METHODS: GABA rho 1 cDNA from human retina was transcribed in vitro to obtain sense rho 1 mRNA, which was microinjected into Xenopus oocytes. Two-electrodes voltage clamp technique was performed to record GABA-induced currents. RESULTS: Expressed receptors were found to have similar properties to GABAc receptors characterized in the retina. Cl-currents induced by GABA were blocked by picrotoxin instead of bicuculline. GABA-induced currents reversed at -19 +/- 2.5 mV, and EC50 was 3.3 mumol/L. Zn++ modulated GABA-induced currents with an IC50 = 9.6 mumol/L. Ni++, Cu++ and Cd++ inhibited GABA rho 1 obviously, too. Their rank order of potency was Zn++ > Ni++ > Cu++ > Cd++. CONCLUSION: Zinc (10 mumol/L) inhibited GABA induced currents in a competitive manner, and its action was sensitive to extracellular pH. Site-directed mutagenesis revealed that substitution of a single histidine residue (H44 and H48) failed to affect zinc sensitivity. PMID- 11862607 TI - G1 arrest and relative protein expressions in mouse thymocytes induced by whole body X-ray irradiation. AB - OBJECTIVE: To investigate the molecular regulation of G1 arrest of mouse thymocytes induced by ionizing radiation. METHODS: Cell cycle was analyzed by flow cytometry (FCM) following staining of cells with propidium iodide. Fluorescent staining and flow cytometry analysis were employed for measurement of protein expression. RESULTS: It was demonstrated that G1 phase of mouse thymocytes increased significantly at 12 h after whole body irradiation (WBI) with the doses of 0.5, 1.0 and 2.0 Gy, and at 24 h following 2.0 Gy exposure, measured by FCM. In the time course experiment, it was found that G1 phase of thymocytes increased significantly at 4 h, reached a peak level at 24 h and came down toward 48 h after WBI with 2.0 Gy X-rays. The results also showed that after 2.0 Gy exposure, the expression of proteins in mouse thymocytes increased significantly from 1 h to 8 h for p53, for p21 from 4 h to 48 h, and for MDM2 at 4 h and 8 h, measured by FCM. But no change was found for GADD45 protein expression. CONCLUSION: These results suggest that G1 arrest could be induced by a single dose of 0.5 Gy, 1.0 Gy or 2.0 Gy, and its molecular control might be established through the p53-p21 pathway. PMID- 11862608 TI - Definition and measurement of child malnutrition. AB - Child Malnutrition has been defined or described in many ways. To summarize the common points, child malnutrition may be defined as a pathological state resulting from inadequate nutrition, including undernutrition (protein-energy malnutrition) due to insufficient intake of energy and other nutrients; overnutrition (overweight and obesity) due to excessive consumption of energy and other nutrients; deficiency diseases due to insufficient intake of one or more specific nutrients such as vitamins or minerals. In the assessment of child malnutrition, a large number of measures are available. Among these measuring methods, Z-score, growth charts, weight-for-length index (WLI), height standard weight, body mass index, skin fold thickness and clinical grading diagnosis standard for child malnutrition are selected for discussion. The same set of anthropometry data of a group of preschool children was analyzed by using different methods. The results showed that it was very difficult to determine the nutritional state of children using a single index. For a given group of children, Z-score method is recommended to observe their height, weight and weight for height at the same time in order to have a comprehensive understanding. PMID- 11862609 TI - Expression of human vascular endothelial growth factor receptor Flt-1 extracellular domain 1-3 loop cDNA in Pichia pastoris, purification of the expressed product and detection of its biological activity. AB - OBJECTIVE: To express human vascular endothelial growth factor receptor Flt-1 extracellular domain 1-3 loop cDNA in Pichia. pastroris, and to purify the expressed product and detect its biological activity. METHODS: By inserting human Flt-1 (1-3 loop) cDNA coding 316 amino acid residues into Pichia pastoris expression vector pPIC9K containing AOX1 promoter and the sequences of alpha secreting signal peptides, a recombinant expression plasmid pPIC9K/Flt-1 (1-3) was constructed and transformed to yeast host strain GS115, then His+ Muts phenotype transformant was screened out and cultured in flasks, and Flt-1 (1-3) was expressed under the induction of 1% methanol. RESULTS: SDS-PAGE showed that after being induced with 1% methanol for 4d, the expressed product existed in supernatant in the form of soluble molecule and contained 60% of total protein expressed. Western blot showed good antigenicity and specificity of expressed product. After being purified by CM-Sepharose FF and Sephacryl S-100 chromatography, the purity of the expressed product reached above 90%. Biological assay proved that the expressed product could bind to hVEGF165 and inhibit the proliferation of HUVEC stimulated by hVEGF165. CONCLUSION: Human vascular endothelial growth factor receptor Flt-1 extracellular domain 1-3 loop was successfully expressed. The study lays a foundation for further application of the expressed product in the treatment of vasoformation related diseases, such as tumor and diabetic retinopathy. PMID- 11862610 TI - Expression of human vascular endothelial growth factor (VEGF165) in Pichia pastoris and its biological activity. AB - OBJECTIVE: To express human vascular endothelial growth factor (hVEGF165) cDNA in Pichia pastroris, purify the expressed product and detect the biological activity of it. METHODS: By inserting hVEGF165 cDNA coding 165 amino acid residues into Pichia pastoris expression vector pPIC9K containing AOX1 promoter and the sequences of alpha secreting signal peptides, a recombinant expression plasmid pPIC9K/hVEGF165 was constructed and transformed to yeast host strain KM71, then multiple-copy insert transformants were screened out and cultured in flasks, and hVEGF165 was expressed under the induction of 1% methanol. RESULTS: SDS-PAGE showed that after being induced with 1% methanol for 4 d, the expressed product existed in supernatant in the form of soluble molecule and contained 60% of total protein expressed. Western blot showed good antigenicity and specificity of expressed product. After being purified by Heparin-Sepharose CL6B affinity chromatography, the purity of expressed product reached above 90%. Biological assays proved that the expressed product could stimulate the proliferation of HUVEC. CONCLUSION: hVEGF165 was successfully expressed. The study opened up a wide prospect for the application of VEGF165 in the prevention and treatment of ischemic heart disease and other tissue ischemic diseases such as secondary arterial occlusion in limbs. PMID- 11862611 TI - Effects of cadmium on rat sperm motility evaluated with computer assisted sperm analysis. AB - OBJECTIVE: To study effects of cadmium on rat sperm motility evaluated with computer assisted sperm analysis. METHODS: Different doses of cadmium chloride (0.2, 0.4, 0.8 mg Cd/kg BW) were administrated i.p. to adult male Sprague-Dawley rats. Control animals received the same volume of 0.9% NaCl solution. After 7 days, the rats were sacrificed with their testes removed. A part of one testis was used for testicular sperm head counts and daily sperm production observation. The motility of spermatozoa obtained from cauda epididymides using the "diffusion" method was measured by computer assisted sperm analysis (CASA). RESULTS: The sperm head counts and daily sperm production decreased significantly in the high dose group. The motility of spermatozoa in the middle dose group was reduced significantly. No motile sperm was found in the high dose group. The results suggest that germinal epithelium was impaired irreversibly in a short time to produce toxic effects on spermatogenesis at high cadmium doses. CONCLUSION: Cadmium may reduce sperm motility at a dose far below the dose affecting sperm production at this time point. The motility of sperm is an early and sensitive endpoint for the assessment of cadmium toxicity on male reproduction. PMID- 11862612 TI - Dietary intake of some essential micronutrients in China. AB - OBJECTIVE: Based on the data collected in the 1992 national nutrition survey in China, the food consumption and nutrients intake were calculated, and the consumption of some micronutrients was evaluated. METHOD: Dietary data were obtained by using a three days' inventory change plus food weighing in combination with 24 hours recall method for three consecutive days. The food consumption and nutrients intake were calculated in accordance with the Chinese food composition table. The consumption of some micronutrients was evaluated in reference to the Chinese RDAs. RESULTS: The average intakes of niacin, ascorbic acid and vitamin E were sufficient, whereas that of zinc, selenium and thiamin were between 80% and 90% RDAs. The consumption of calcium, retinol equivalent and riboflavin was low compared with the Chinese RDAs. Calcium was the most insufficient nutrient in Chinese diet. CONCLUSION: Food fortification seems necessary for improving the nutritional status of some micronutrients in China. PMID- 11862613 TI - Hepatoxic alterations induced by inhalation of trichlorethylene (TCE) in rats. AB - OBJECTIVE: Trichlorethylene (TCE) is one of the most potent organic unsaturated solvents being used in dry cleaning, metal degreasing, thinner for paints varnishes and electroplating, etc. and has been reported to be a hepatoxicant through oral and dermal exposure. However, its inhalation toxicity data is very limited in the literature due to the fact that the exposure levels associated with these effects were usually not reported. Hence, inhalation toxicity study was carried out for hepatotoxic studies. METHODS: Inhalation toxicity studies was carried out by exposing rats to TCE for 8, 12 and 24 weeks in a dynamically operated whole body inhalation chamber. Sham treated control rats were exposed to compressed air in the inhalation chamber for the same period. RESULTS: Significant increase in liver weight (liver enlargement) appeArance of necrotic lesion with fatty changes and marked necrosis were observed after longer duration (12 and 24 weeks) of TCE exposure. The lysosomal rupture resulted in increased activity of acid and alkaline phosphatase alongwith reduced glutathione content and total increased sulfhydryl content in liver tissue. CONCLUSION: TCE exposure. through inhalation route induces hepatotoxicity in terms of marked necrosis with fatty changes and by modulating the lysosomal enzymes. PMID- 11862614 TI - Pulmonary toxicity of a formulated preparation of fenvalerate in rats subchronically exposed by nose only inhalation for 90 days. AB - OBJECTIVE: The pulmonary toxicity of a commercially available formulated preparation of Fenvalerate (Fen), a synthetic pyrethroid has been studied in rats following subchronic nose only inhalation exposure route. METHOD: Adult male rats were exposed to Fen for 4 h/day, 5 days a week for 90 days by using Flow Past Dynamic Nose only Inhalation Chamber. RESULTS: Fen exposed rats showed a significant increase in enzymatic activities of lactate dehydrogenase (LDH), acid phosphatase (ACP), alkaline phosphatase (ALP) and gamma-glutamyl transferase (gamma-GT) which are considered as biochemical indicators of pulmonary damage. The concomitant histopathological examination of Fen exposed rats' lung revealed inflammatory changes viz., influx of mononuclear cells admixed with a few giant cells in alveolar lumen, hypetrophied bronchiolar and alveolar epithelial lining cells and presence of edematous fluid in alveolar lumen alongwith congested parenchymatous blood vessels. CONCLUSION: These results for the first time indicate the pulmonary toxic effects of a commonly used formulated Fen preparation by using rat model and nose only inhalation as the route of exposure. PMID- 11862615 TI - The effect of antihypertensive treatment on cardiovascular events--a meta analysis of four clinical trials in China. AB - OBJECTIVE: The pooled results of four large-scale clinical trials of antihypertensive treatments carried out in China were assessed, with an overall study population of 10,457 patients. METHODS: The fixed effect model was used for data analysis. RESULTS: A pooled analysis at the individual level showed that the average time of treatment was 3.4 years, after which an average reduction in blood pressure of 9/4 mmHg was found. The meta-analysis showed that stroke was reduced by 36% (95% CI 25%-46%, P < 0.001), cardiovascular death and total deaths were reduced by 22% and 20% respectively (P < 0.05) and total cardiovascular events were significantly reduced by 33% (P < 0.001). However, the reduction in coronary heart disease (CHD) was much smaller and was not statistically significant (P > 0.05), whereas the significant reduction in stroke was epidemiologically expected. CONCLUSION: The benefit of antihypertensive treatment obtained was mostly due to a reduction in the incidence of stroke. PMID- 11862617 TI - Reaction to the emergence of BSE in the UK: what was done and what perhaps might have been done better. PMID- 11862616 TI - PrPC has nucleic acid chaperoning properties similar to the nucleocapsid protein of HIV-1. AB - The function of the cellular prion protein (PrPC) remains obscure. Studies suggest that PrPC functions in several processes including signal transduction and Cu2+ metabolism. PrPC has also been established to bind nucleic acids. Therefore we investigated the properties of PrPC as a putative nucleic acid chaperone. Surprisingly, PrPC possesses all the nucleic acid chaperoning properties previously specific to retroviral nucleocapsid proteins. PrPC appears to be a molecular mimic of NCP7, the nucleocapsid protein of HIV-1. Thus PrPC, like NCP7, chaperones the annealing of tRNA(Lys) to the HIV-1 primer binding site, the initial step of retrovirus replication. PrPC also chaperones the two DNA strand transfers required for production of a complete proviral DNA with LTRs. Concerning the functions of NCP7 during budding, PrPC also mimices NCP7 by dimerizing the HIV-1 genomic RNA. These data are unprecedented because, although many cellular proteins have been identified as nucleic acid chaperones, none have the properties of retroviral nucleocapsid proteins. PMID- 11862618 TI - Neuropathology of variant Creutzfeldt-Jakob disease. AB - The neuropathological features human prion diseases comprise spongiform change, neuronal loss, astrocytic and microglial proliferation and the accumulation of the abnormal isoform of prion protein (PrPRES) in the central nervous system. Variant Creutzfeldt-Jakob disease (CJD) is a novel human prion disease which appears to result from infection by the bovine spongiform encephalopathy (BSE) agent. The neuropathology of variant CJD shows morphological and immunocytochemical characteristics distinct from all other types of human prion disease, and is characterised by abundant florid and cluster plaques in the cerebrum and cerebellum, and widespread accumulation of PrPRES on immunocytochemistry. Spongiform change is most marked in the caudate nucleus and putamen, and the thalamus exhibits severe neuronal loss and gliosis, which is most marked in the posterior nuclei and correlates with the areas of high signal seen in the posterior thalamus on MRI examination of the brain. Western blot analysis of PrPRES on frozen brain tissue in variant CJD tissue shows a uniform isotype, with a glycoform ratio distinct from sporadic CJD. PrPRES accumulation is widespread in lymphoid tissues in vCJD. All cases of variant CJD are methionine homozygotes at codon 129 of the PrP gene. Histological and biochemical techniques will be required to identify cases of 'human BSE' in individuals who are MV or VV at codon 129 of the PrP gene. Continued surveillance is required to investigate this possibility in the UK and other countries where BSE has been reported. PMID- 11862619 TI - Structure and assembly properties of the yeast prion Ure2p. AB - The non-Mendelian phenotype [URE3] is due to a transmissible conformational change of the protein Ure2. The infectious protein form of Ure2p has lost its function and gained the capacity to transform the active form of the protein into an inactive form. The molecular basis of this conversion process is unknown. There are however indications that the conformational changes at the origin of the propagation of the inactive form of Ure2p in yeast cells are similar to those at the origin of the transition of PrPC into the scrapie-associated PrPSc form of the protein. To better understand the nature of the conformational changes at the origin of prion propagation, we have purified, characterized biochemically, examined the assembly properties and solved the crystal structure of Ure2p. Our data are presented below and a number of conclusions dealing with the molecular basis of the conversion of soluble Ure2p into its amyloid-forming state are derived. PMID- 11862620 TI - Predicting the size of the vCJD epidemic in France. AB - More than 5 years after the description of the first cases of variant Creutzfeldt Jakob disease (vCJD), there is still great uncertainty about the size of the vCJD epidemic in the United Kingdom (UK), although the most recent predictions based on statistical modelling are more optimistic than the previous ones. The number of vCJD cases in France is far too small to attempt any direct modelling of the vCJD epidemic in the French population. Comparative assessment of the level of exposure to the bovine spongiform encephalopathy (BSE) agent in the UK and France could help to estimate the size of the vCJD epidemic in France. Data on imports of beef products from the UK between 1985 and 1996, BSE epidemic in French cattle, and travel of French people to the UK suggest that the French population was much less exposed to the BSE agent than the UK population. The France/UK ratio of vCJD incidence is currently approximately equal to 0.05. Further studies are needed to estimate accurately the exposure ratio between UK and France and to examine whether comparative data about exposure and incidence are fully consistent. The temporal pattern of exposure in UK and France, and possible differences in exposure to high risk bovine tissues because of food habits or risk reduction measures should be carefully considered. PMID- 11862621 TI - The transmission dynamics of BSE and vCJD. AB - The bovine spongiform encephalopathy (BSE) epidemic in cattle has had a huge economic impact on the agricultural industries across Europe. Furthermore, scientific evidence now strongly supporting a link between a new variant of Creutzfeldt-Jakob disease (vCJD) and consumption of BSE-infected animals has further heightened the need both to understand the transmission of these new diseases and to improve control measures to protect public health. In this paper we review work undertaken by our group using epidemiological models to understand the transmission dynamics of BSE and vCJD. We present new estimates of the future number of cases of BSE and the number of infected animals slaughtered for consumption for Great Britain, and summarise similar analyses undertaken for Northern Ireland, Ireland, Portugal and France. We also consider the epidemiological determinants of the future course of the vCJD epidemic, including the age and genetic characteristics of the confirmed cases, and present predictions of future case numbers. PMID- 11862622 TI - New in vivo and ex vivo models for the experimental study of sheep scrapie: development and perspectives. AB - Sheep scrapie is a prototypical transmissible spongiform encephalopathy (TSE), and the most widespread of these diseases. Experimental study of TSE infectious agents from sheep and other species essentially depends on bioassays in rodents. Transmission of natural sheep scrapie to conventional mice commonly requires one or two years. In an effort to develop laboratory models in which investigations on the sheep TSE agent would be facilitated, we have established mice and cell lines that were genetically engineered to express ovine PrP protein and examined their susceptibility to the infection. A series of transgenic mice lines (tgOv) expressing the high susceptibility allele (VRQ) of the ovine PrP gene from different constructs was expanded. Following intracerebral inoculation with natural scrapie isolates, all animals developed typical TSE neurological signs and accumulated abnormal PrP in their brain. The survival time in the highest expressing tgOv lines ranged from 2 to 7 months, depending on the isolate. It was inversely related to the brain PrP content, and essentially unchanged on further passaging. Ovine PrP transgene expression thus enhanced scrapie disease transmission from sheep to mice. Such tgOv mice may bring new opportunities for analysing the natural variation of scrapie strains and measuring infectivity. As no relevant cell culture models for agents of naturally-occurring TSE exist, we have explored various strategies in order to obtain stable cell lines that would propagate the sheep agent ex vivo without prior adaptation to rodent. In one otherwise refractory rabbit epithelial cell line, a regulable expression of ovine PrP was achieved and found to enable an efficient replication of the scrapie agent in inoculated cultures. Cells derived from sheep embryos or from tgOv mice were also used in an attempt to establish permissive cell lines derived from the nervous system. Cells engineered to express PrP proteins of a specified sequence may thus represent a promising strategy to further explore, at the cellular level, various aspects of TSE diseases. PMID- 11862623 TI - [Cell culture models of transmissible spongiform encephalopathies]. AB - Cell cultures represent versatile and useful experimental models of transmissible spongiform encephalopathies. These models include chronically prion infected cell lines, as well as cultures expressing variable amounts of wild-type, mutated or chimeric prion proteins. These cultures have been widely used to investigate the biology of both the normal and the pathological isoform of the prion protein. They have also contributed to the comprehension of the pathogenic processes occurring in transmissible spongiform encephalopathies and in the development of new therapeutic approaches of these diseases. PMID- 11862624 TI - BSE infection of the small short-lived primate Microcebus murinus. AB - Eleven Microcebus murinus (lemur) primates were intracerebrally or orally infected by bovine spongiform encephalopathy (BSE) or macaque-adapted BSE (MBSE) brain homogenates. In many BSE and MBSE infected lemurs, but not in animals inoculated with normal bovine brain, persistent behavioral changes occurred as early as 3 months, and neurological signs as early as 13 months after infection. Immunohistochemical examination of animals sacrificed during the incubation period revealed an abnormal accumulation of 'prion' protein (PrP) in the intestinal wall, intestinal nervous plexus, mesenteric lymph nodes and spleen, and in the clinical stage, also in the brain. In MBSE-inoculated animals, proteinase K resistance of the PrP (PrPres) was confirmed by Western blot in the spleen and the brain. Obvious signs of neurodegeneration were observed in all infected animals characterized by hyperaggregated and paired-helical filaments immunoreactive Tau proteins, beta 42-amyloid plaques and astrogliosis. Additionally, PrPres was present in the ganglion cells of the retina in diseased animals after either intracerebrally or oral infection by the BSE or MBSE agent. These results show that the microcebe is susceptible to the BSE infectious agent via intracerebral and oral routes with comparatively short incubation periods compared to simians, and could be a useful animal model to study the pathophysiology of disease transmission in primates. PMID- 11862625 TI - Inactivation of the BSE agent. AB - In the studies carried out so far, the BSE agent has proved to be just as resistant as other TSE agents to inactivation by procedures such as autoclaving or exposure to sodium hydroxide that are effective with conventional microorganisms. However, in common with other TSE agents, the BSE agent appears to be effectively inactivated by exposure to sodium hypochlorite solutions containing high levels of available chlorine. Not surprisingly, the BSE agent has been found to survive at least some of the rendering processes that were used to process tissues discarded by abattoirs in the EU during the early 1980s. Despite the survival of BSE infectivity after autoclaving or exposure to sodium hydroxide, it is known that combining these procedures results in a very reliable degree of inactivation for TSE agents generally. The combination of heat and alkali has also been shown to be effective with a mouse-passaged strain of BSE agent, even at a temperature of only 100 degrees C for a minute. Also, in carrying out BSE-spiked validation studies relating to the safety of bone-derived gelatin, it has also been found that the exposure of acid-treated bone (which is free from any obvious remains of fatty or proteinaceous tissue) to 0.3 M sodium hydroxide for two hours knocks out any residual BSE infectivity. PMID- 11862626 TI - From stem cells to prion signalling. AB - A strategy based upon the introduction of an adenovirus-SV40 plasmid into multipotential cells was designed to immortalize clones displaying properties of lineage stem cells. The murine 1C11 cell line behaves as a neuroepithelial progenitor. Upon appropriate induction, almost 100% of 1C11 precursor cells develop neurite extensions and convert into either serotonergic or noradrenergic neurons. The two mutually exclusive neuronal programs are autoregulated by serotonergic or adrenergic receptors. PrPc is constitutively expressed by 1C11 cells. Antibody-mediated cross-linking of PrPc promotes the dephosphorylation of the tyrosine kinase Fyn associated to a Fyn kinase activation. The coupling of PrPc to Fyn is dependent on caveolin-1. It is restricted to the fully differentiated serotonergic or noradrenergic cells and occurs mainly at neurites. Thus, PrPc may represent a signal transduction protein which may fine-tune neuronal functions. Since the 1C11 stem cell supports prion replication, it may provide a tool to investigate whether PrPSc accumulation interferes with PrPc signalling activity. PMID- 11862628 TI - CMS drops employment requirement for 'incident-to' billing by pharmacists. PMID- 11862627 TI - Anthrax vaccine offered, but not recommended, as option to treat infection. PMID- 11862629 TI - Cytokine blocker licensed for relief of rheumatoid arthritis symptoms. PMID- 11862630 TI - ACIP extends delay in pneumococcal immunization of healthy children. PMID- 11862631 TI - Study finds inappropriate medication use by community-dwelling elderly. PMID- 11862633 TI - USP launches dietary supplement certification program. PMID- 11862632 TI - Methylphenidate isomer approved by FDA. PMID- 11862634 TI - Cilostazol. PMID- 11862636 TI - Summary of the executive session on emergency preparedness and the pharmaceutical supply chain. PMID- 11862635 TI - Colesevelam hydrochloride. PMID- 11862637 TI - Implications of prion-induced diseases for animal-derived pharmaceutical products. AB - The implications of prion-induced diseases for the use of medications that theoretically could harbor the infectious pathogens are discussed. Prions have been identified as protein particles that lack nucleic acids. There is evidence that prions cause the transmissible neurodegenerative diseases known as transmissible spongiform encephalopathies. Of these diseases, bovine spongiform encephalopathy (BSE) and the human spongiform encephalopathy to which it has been linked, new variant Creutzfeldt-Jakob disease (CJD), have generated the most attention. The first cases of new variant CJD appeared in Britain in the mid 1990s. Ingestion of prion-infected beef remains the only known cause of new variant CJD. No cases of BSE or new variant CJD have been documented in the United States. The time from exposure to the development of clinical sequelae appears to be about 10 years. The median duration of illness is 14 months, and the outcome is invariably death. There is no treatment; currently the only available approach is prevention. There is no reliable method of predicting the number of new cases that might occur because of lack of definitive information on the efficiency of transmission from animals to humans and the number of people currently infected and at risk for infection. The infectivity of medications and plasma fractionation products containing material from cattle with BSE is unknown, but the risk is believed to be very low. No cases of such transmission have been identified. Guidelines to keep the risk of transmission via medications low have been promulgated by FDA, and further research is warranted. There have been no reports of medications or plasma fractionation products being contaminated with the prions that cause new variant CJD. Ongoing vigilance and research are appropriate, however. PMID- 11862638 TI - Compliance with safe practices for preparing parenteral nutrition formulations. AB - Compliance with the guidelines of the National Advisory Group on Standards and Practice Guidelines for Parenteral Nutrition (NAG) was studied. A questionnaire based on the NAG guidelines was developed to identify common practices used in the preparation of parenteral nutrition formulations in a variety of health care settings. The questionnaire was mailed in January 2000 to a national sample of 1445 pharmacists. There were a total of 633 usable responses, for a net response rate of 45.2% (based on 1400 deliverable surveys). Almost 80% of the respondents were employed in an urban setting, and 64.3% worked in hospitals. Relatively high compliance was reported for guidelines related to filtration (86.9% and 85.9%), verification of product stability and compatibility by a pharmacist or computer program (86.4%), evaluation of the aseptic technique of persons preparing sterile products (81.4%), and having a pharmacist assess formulations to determine whether the contents are within acceptable standard ranges (78.8%). Low compliance was reported for guidelines related to the inclusion of dosing weight on the label (15.4%), the use of amount per day as the unit of measure for labeling ingredients (26.1%), direct pharmacist-to-pharmacist communication about prescriptions when patients are transferred among health settings (33.4%), and daily in-process or end-product quality assurance testing of formulations. The NAG guidelines on parenteral nutrition formulations with which pharmacists' practice was most discrepant were the use of amount per day as the unit of measure for labeling ingredients, inclusion of the patient's dosing weight on the label, direct pharmacist-to-pharmacist communication when patients are transferred, and daily quality assurance testing. PMID- 11862639 TI - Monitoring method for surface contamination caused by selected antineoplastic agents. AB - A method of evaluating surface contamination caused by selected antineoplastic agents was studied. The antineoplastic agents tested were cyclophosphamide, ifosfamide, doxorubicin hydrochloride, fluorouracil, and paclitaxel. Each agent was reconstituted and prepared as a stock solution. A 0.1-mL portion of each solution was spread evenly over a 600-cm2 area of a stainless steel surface, a resin countertop surface, and a vinyl flooring surface. After drying, the surfaces were wiped with each of two types of commercially available wiping materials (What-man no. 42 filters and Kimberly-Clark Kimwipes). A blend of methanol, acetonitrile, and buffered water was used both as the wetting agent for wiping the surfaces and as a desorbing solution. The desorbate was analyzed for drug concentration by reverse-phase high-performance liquid chromatography (HPLC). Mean +/- S.D. percent total recovery ranged from 72.4% +/- 17.6% to 95.3% +/- 2.9% for the vinyl surface wiped with filters, 91.5% +/- 5.4% to 104.7% +/- 0.8% for the resin surface wiped with filters, 73.9% +/- 2.3% to 95.3% +/- 1.7% for the stainless steel surface wiped with filters, and 18.2% +/- 1.4% to 372.8% +/- 8.0% for the stainless steel surface wiped with Kimwipes. Results were best for ifosfamide and cyclophosphamide. Kimwipes were deemed ineffective for this monitoring method because an ingredient interfered with the quantitative analytical tests. A wipe-sampling, desorption, and HPLC method for monitoring surface contamination by selected antineoplastic agents was sufficiently accurate and sensitive to evaluate surfaces typically found in both the pharmacy and drug administration areas of oncology treatment facilities. PMID- 11862640 TI - Perspectives on the attributes and characteristics of pharmacy executives. PMID- 11862641 TI - ASHP statement on the role of health-system pharmacists in counterterrorism. PMID- 11862642 TI - Computerized physician order entry: challenges and opportunities. PMID- 11862643 TI - When is the best time to hire an associate? AB - Hiring an associate is a natural step in the growth of a dental practice. However, it is a choice not to be made lightly or based on vague perceptions and expectations. A dentist wishing to bring another dentist into the practice needs to consider concerns such as the business of the operative schedule, personal and professional wishes in regard to sharing the workload, and how future practice growth or retirement will impact the choice of an assistant. PMID- 11862644 TI - Managing malpractice liability: tips to limit your risk. AB - Perceived poor communication and poor documentation are the primary culprits in litigation against dentists. Better communication can be attained by making strong efforts to involve the patient in the decision-making process when proposing treatment, and recording those discussions in the patient's record. Records should be meticulously kept, for legibility, accuracy, and objectivity protect a dentist from liability. The patient's medical history should also be a part of the record. PMID- 11862645 TI - Provide a service ... then get paid. AB - Receiving payment at the time of dental treatment may be a new--even foreign- concept to dentists today. But more and more practices are finding it necessary to enforce such payment policies to keep a practice viable. Sometimes, it's just a matter of establishing policies within a practice; in other cases, it pays to enlist the services of a dental financing company for patient collections. PMID- 11862646 TI - Tax-advantaged investing: a wise choice. AB - Your investment strategy should be just that--a plan to make the most of your assets. Considering the tax advantages and disadvantages help you stretch your investments and take full advantage of stocks, mutual funds, and other investments. PMID- 11862647 TI - Education planning for today. AB - Parents have several options when it comes to saving for a child's college education. The choices vary as to how early you begin; mutual funds, education IRAs, and prepaid tuition plans are among the choices when your children are young. If you don't start saving until your child is in high school, your choices can include loans and financial aid. PMID- 11862648 TI - Treat patients' concerns as well as their oral health. PMID- 11862649 TI - Improve your chances of incorporating change. AB - Successfully initiating change in a dental practice means understanding that several management issues need to be recognized: involvement of staff in the process of change; clear goals and a vision that are understood by everyone; leadership that stays involved; and the celebration of small wins and short-term goals. PMID- 11862650 TI - General anaesthesia for dental treatment in a hospital setting with critical care facilities. A letter from the Department of Health, 31 May 2001. PMID- 11862651 TI - Acupuncture TMD and facial pain. PMID- 11862652 TI - Association of Dental Anaesthetists. Summer Scientific Meeting Stirling, Scotland 8-9 June, 2001. ADA meeting report. AB - Local anaesthesia in addition to general anaesthesia can alleviate pain and distress caused by painful procedures in community dentistry for special needs patients. Further studies are needed to confirm these results in a larger group of patients. PMID- 11862653 TI - Hot topics in AIDS therapeutics. PMID- 11862654 TI - Research developments in HIV/AIDS. PMID- 11862655 TI - Drug-resistant HIV on rise as medicines misused. PMID- 11862656 TI - Management issues in patients coinfected with hepatitis C virus and HIV. AB - Coinfection with HIV accelerates the progression of hepatitis C toward advanced liver disease. Low CD4+ cell counts may result in false-negative results on all diagnostic tests except hepatitis C virus (HCV) RNA assays, which are the gold standard for viral replication. First-line management of HIV-HCV--coinfected patients should be optimization of HAART, because low CD4+ cell counts have been associated with greater fibrosis. In addition, agents used to treat hepatitis C may lower CD4+ cell counts and hemoglobin levels. Long-acting interferons offer the promise of better sustained HCV response in HIV-HCV coinfection. PMID- 11862657 TI - Hepatitis C management in HIV infection--where is the controversy? PMID- 11862658 TI - Assessing HIV-associated dementia: modified HIV dementia scale versus the Grooved Pegboard. AB - The HIV dementia scale (HDS) previously has been shown to be sensitive in identifying persons with HIV-associated dementia (HAD). We aimed to develop a simpler version of the HDS--the Modified HDS (M-HDS)--which would be appropriate for use by nonneurologists. We sought to determine whether the M-HDS and the widely used Grooved Pegboard (GP) were comparable in identifying and staging the severity of dementia in HIV-seropositive persons. Data from 455 HIV-seropositive persons were analyzed, and patients were separated into 2 groups: demented (n = 144) and nondemented (n = 311), where the diagnosis was established by a single neurologist. Of the 144 with HAD, 13 had severe dementia, 55 had moderate dementia, 51 had mild dementia, and 25 had minor cognitive/motor disorder. We assessed the relationship between dementia severity and score on the M-HDS and GP (nondominant hand). Results showed both the M-HDS (z = -4.32; P < .05) and the GP nondominant (z = -4.08; P < .05) differentiated demented from nondemented persons. Furthermore, the M-HDS (z = -2.95; P < .05) and GP nondominant (z = 2.68; P < .05) discriminated equally well the stages of dementia severity. A cutoff score of 7.5 on the M-HDS maximized its sensitivity (70%) and specificity (71%). The M-HDS and the GP (nondominant) were found to be equally sensitive and specific in categorizing and staging HAD. However, the M-HDS requires no equipment and is simpler to administer; so it may be more useful for screening by nonneurologists. PMID- 11862659 TI - Screening for the AIDS dementia complex. PMID- 11862660 TI - One more day. PMID- 11862661 TI - [Challenges in chemistry: color reactions, cycloadditions and cycloreversions]. AB - New colour reactions for the identification of drug compounds have been devised. For example, butazolidine is oxidized with permanganate in basic medium to azobenzene and butyric acid; after reduction and coupling, the former is transformed to a red azodye. On nitration, gluthetimide gives the p-nitrophenyl derivative and a second nitro group is substituted at position 3. With hydroxylamine in basic medium, this o-dinitro compound reacts to furnish a violet colour when the dinitro derivative is reduced to the ortho-quinoidal dialkali metal salt. Dipolar cycloadditions of BNO, ANO and DPNI to norbornane- and cyclohexane-condensed 1,3-oxazine dipolarophiles have been utilized to synthesize adducts containing 1,2,4-oxadiazole and -triazole rings. The norbornene double dipolarophiles contain both C=N and C=C bonds; for steric reasons, ketene formed in situ adds primarily to the C=C. The addition of dipoles with high steric demands can lead to a change in conformation. By means of retro Diels-Alder reactions, hetero- mono-, bi- and tricyclic compounds may be synthesized. With bifunctional reagents, norbornene amino acids and hydrazides are transformed to condensed cyclic derivatives; when these are heated, cyclopentadiene is cleaved off to give mostly new target compounds. PMID- 11862662 TI - [Application possibilities of dynamic laser light scattering photometry for the examination of poly(ethylene glycol) suppositories]. AB - Status epilepticus is one of the most common neurologic emergencies in children, adolescents, and young adults. The advantage of diazepam suppository in medicinal therapy appears mainly in treatment of childhood epilepsy. A hydrophilic suppository base was investigated in solution by dynamic light scattering and results were compared to those of the membrane diffusion experiments measuring diazepam solubilization. According to the dynamic laser light scattering photometric measurements, the good solubilization effect of the macrogol mixture (5% Polysorbatum 20, 10% Macrogolum 400, 85% Macrogolum 1540) is explained by the formation of small, tight micellas. As the Polysorbatum 20 tenside resulted in the significant decrease (p < 0.05) of the formation of great micellas, its use led to the formation of small, tight, almost monodisperse micellas of 40-50 nm in the aqueous solution of the macrogol mixture. PMID- 11862663 TI - [Structural diversity of ecdysteroids of Lychnis flos-cuculi]. AB - Eleven ecdysteroids have been isolated from Lychnis floscuculi; we are the first who report eight ecdysteroids of the eleven compounds in this plant. Two of these ecdysteroids, dihydrorubrosterone and 20-hydroxyecdysone 3-acetate are newly discovered natural products. The success of isolation of these new ecdysteroids has been based on the use of separation methods in a proper order; these separation procedures were completing each others. At the beginning steps of isolation simple separation methods were used, such a solvent-solvent distribution and fractionated precipitation. Two third of the contaminants were removed thereby. High capacity low resolution methods were used then, such as classical adsorption column chromatography and preparative thin-layer chromatography. The major component (20-hydroxyecdyssone) and certain minor ecdysteroids (polypodine B and rubrosterone) were isolated in pure form here. Purification of the further minor components (poststerone, 2-deoxy-20 hydroxyecdysone, vitikosterone E, dihydrorubrosterone, makisterone A, taxisterone, 20-hydroxyecdysone 2-acetate, 20-hydroxyecdysone 3-acetate) required HPLC and other absorption chromatographic methods. Our recent separation scheme means a generally applicable guiding principle for isolation of any plant ecdysteroid, major and minor alike. Structural identification of the known ecdysteroids was based on their spectral data and that of their literature information. Structural elucidation of 20-hydroxyecdysone 3-acetate was done by the help of a standard component prepared by acetylation of 20-hydroxyecdysone. From the mixture of seven acetates the corresponding compound (20-hydroxyecdysone 3-acetate) was isolated, and used for identification. Structural diversity of ecdysteroids of Lychnis flos-cuculi is evaluated, and a tentative explanation is introduced for the formation and biosynthesis of the versatility of phytoecdysteroids. PMID- 11862664 TI - [A novel pyridazino-fused ring system: synthesis of pyridazino[3,4-b]diazepam]. AB - As an analogue of pyridazino-fused ring systems with pharmacological activities, the novel pyridazinol[3,4-b][1,5]diazepine ring system was prepared. The synthetic pathway includes three steps from 4 5-(N-benzyl-N-3-hydroxypropyl)amino derivative which is easily available through nucleophilic substitution reaction of the known 4,5-dichloro-2-methyl-6-nitro-3(2H)-pyridazinone (2) with N-benzyl-N (3-hydroxypropyl)amine. In the first step, compound 4 was treated with thionyl chloride to give the chloropropyl derivative 5. In the second step, a Bechamp reduction was carried out with Fe in acetic acid to obtain the amino compound 6, and finally the ring closure reaction of 6 was performed in N,N-dimethylformamide in the presence of potassium carbonate at 110 degrees C for 40 hours. In this way the bicyclic compound 7 could be isolated in 48% yield. PMID- 11862665 TI - [Synthesis of carboline alkaloid analogues]. AB - Hybrid compounds were synthesized combining the structural features of two isomer natural indolalkaloids rutaecarpine (1) and nauclefine (2). These aza bioisosteric analogues are the first representatives of a new heterocyclic ring system. Two alternative reaction routes were developed for the synthesis of pentacyclic compounds (4, 5) in which the key step is the Fischer indolization of the 6-phenylhydrazono-dipyrido[1,2-a;4,3-d]pirimidine-11-ones. In the case of E ring substituted derivatives the synthesis was carried out via preparation and chemical transformation of pyrido[1,2-a]pirimidine-4-ones (14, 15) to 2 substituted-3-aza-rutaecarpines (17-20). Finally, the nucleophilic displacement of the chlorine atom of 2-chloro-3-aza-rutaecarpine (18) by dialkylaminoethylamine provided the 2-amino-substituted derivative (20) having improved physico-chemical properties and increased antitumour activity. The new compounds are characterized by UV, IR, 1H, 13C NMR spectroscopy. PMID- 11862666 TI - [Influence of adrenergic denervation on uterine contractility of pregnant rats in vitro]. AB - The aim of the present study was to investigate the functional changes in the sympathetic nerves in the rat myometrium during pregnancy by electric field stimulation (EFS) in vitro. In our experiments the contractility of the rat myometrium were registered at 5, 10, 15, 18, 20, 22 (term) day of pregnancy, in non pregnant rats and in 6-OH-dopamine pretreated rats. The parameters of EFS were the following: pulse width 0.6 ms, frequency range of nerve stimulation 1-70 Hz, supramaximal voltage 40 V. We concluded that low pulse width and low frequency stimulation is selective for adrenergic nerves. We found that during pregnancy a gradual drop-out of low frequency induced uterine smooth muscle contraction appears. It is surprising that very low frequency stimulation has no effect on the 5th day of pregnancy although the same frequency stimulation causes contraction on the 10th day. By the 22nd day of pregnancy the rat uterus shows the same responsiveness to nerve stimulation as it is detected in 6-OH-dopamine pretreated non pregnant rats. The adrenergic denervation of the pregnant uterus does not mean that it loses the ability to respond to cathecholamines since non synaptic adrenergic receptors exist in the late pregnant rat uterus. PMID- 11862667 TI - [New data on composition of esssential oil from inflorescence of everlasting (Helichrysum arenarium(L.) Moench.)]. AB - We investigated the composition of essential oil obtained from inflorescence of Helichrysum arenarium (everlasting) which is a perennial herb, native in our home and in Middle and South East Europe. We used essential oils obtained from Hungarian and Polish mercantile samples and a plant sample cultivated in Hungary (Soroksar) by steam-distillation. The GC and GC-MS investigation revealed the presence of more than 60 compounds from which 24 were identified. The identified compounds are as follows: linalool, alpha-terpineol, carvone monoterpenes; anethol, anisaldehyde, thymol, carvacrol, eugenol, beta-asarone, butylhydroxyanisole aromatic components; alpha-humulene, beta-caryophyllene, gamma-muurolene, delta-cadinene, copaene, alpha-gurjunene, caryophyllenol, delta cadinol and globulol sesquiterpenes as well as caprilic, pelargonic, caprinic, laurinic acids and methyl palmitate as alkyl carbonic acid and their derivatives. We established that the main components of essential oil of Hungarian and Polish mercantile samples is methyl palmitate (28.5% and 21.7% respectively) but of cultivated sample is caprinic acid (19.8%). Comparing our results of essential oil composition with those obtained from other Helichrysum species it is surprising that the alkyl carbonic acids are the dominant essential oil constitutents of Helichrysum arenarium. PMID- 11862668 TI - [Preformulation study of atenolol-containing solutions.. I. The pH dependence of thermostability]. AB - The stability of atenolol solutions was evaluated under accelerated isothermal degradation conditions at 90 degrees C. A specific and sensitive HPLC method was adapted to study the pH dependence of the stability. The maximum stability of atenolol was achieved at pH 4 with a k value of 1.1 x 10(-3) h-1. The degradation of atenolol followed first-order kinetics at above mentioned conditions. PMID- 11862669 TI - [Quality of life in asthmatics]. AB - Despite the availability of effective treatments asthma is increasing in both prevalence and severity specially in the well developed countries. As a consequence of this a large part of population is affected and the disease has a significant burden to the society. Evaluation of therapeutic effectiveness can be determined by measuring the quality of life. OBJECTIVE: As asthma has a major impact on the patients everyday life, we examined the quality of life of adult asthmatics with a disease specific questionniare (St. George's Respiratory questionniare SGRQ) and a visual analogue scale (VAS). We wanted to find out if the quality of life scores are significant with the results of clinical tests. METHODS: We examined 321 adult asthmatic patients. Patients were randomly selected. The influence of three factors age, sex, and FEV1 on quality of life was investigated. The data was analysed with user SPSS system. RESULTS: The results of the visuala analog scale were the following in female asthmatics 61.38%, in males 63.86%, the VAS results were the following according to the disease severity: mild intermittant 76%, mild 69%, moderate 60%, sever 49%. From the St. George questionniare three component scores were calculated: symptom (304.15 +/- 11.01), activity (580.71 +/- 19.57), impact (842.18 +/- 31.53) and the total score (1727.05 +/- 57.24). When we examined the different scores according to the age we found significant differences with the activity, impact and total score, but could not find significant differences with the symptom scores. The relationship between spirometry and quality of life is not as strong (coefficient of correlation: 0.37) but the majority of patients have high SGRQ scores even those patients whose FEV1 lies within the normal range. CONCLUSIONS: For patients with asthma it is common that one might see improvement in the FEV1 and yet not see an improvement in patients feelings or functioning. It was interesting to see that in spite of their bad conditions patients do accept the chronic nature of asthma, a degree of impairment and regard that as normal, limits in their everyday life, as they adjust their activity so that they can avoid the asthma attacks. PMID- 11862670 TI - [Suckling-induced changes in oxytocin and alpha-melanocyte-stimulating hormone contents of the median eminence and various lobes of the pituitary gland]. AB - Previous reports have implicated that pituitary-derived prolactin (PRL) is secreted from two distinct zones of mammotropes within the anterior lobe (AL). The inner zone (AL-IZ), located adjacent to the NIL, is supposed to be involved in the rapid and massive discharge of PRL from the pituitary gland due to suckling stimulus. Anatomically, the AL-IZ has an intimate contact with the NIL because the blood arriving from the posterior pituitary through the short portal vessels (SPV) baths it first. Based on these facts it would be hypothesized that the locally released and/or produced important compounds, like oxytocin (OXT) and alpha-melanocyte-stimulating hormone (alpha-MSH), can be delivered to the AL-IZ. Therefore, the purpose of this study was to examine the possible local transportation of these hormones into various regions of the pituitary gland (adenohypophysis inner-zone (AL-IZ), outer zone (AL-OZ), intermediate lobe (IL), neural lobe (NL)) and median eminence of lactating rats. We have measured the concentration of OXT and alpha-MSH from tissue samples of nonsuckled and suckled rats using specific RIA-s. There were no changes in the concentration of OXY and alpha-MSH in the AL-IZ and AL-OZ due to suckling stimulus. In contrast, our data provide compelling evidence that OXT is transported into the IL, which can be further increased by suckling stimulus. Our data have shown a lack of local delivery of either alpha-MSH or OXY into the AL that raises serious doubt about their possible role in PRL secretion during suckling stimulus. PMID- 11862671 TI - [Chromatographic methods in the European Pharmacopoeia]. AB - A brief review is presented on the chromatographic separation techniques used in the European Pharmacopoeia (Ph.Eur.). The corresponding general chapters and the application of each technique in the individual monographs are covered. Special attention is paid to the discussion of the efforts of the Ph.Eur. aiming at the improvement of the reproducibility and reliability of the chromatographic methods in the monographs. In addition to showing the present policy of Ph.Eur. in the application of chromatographic techniques for different analytical purposes the paper also gives an overview on the changes to be expected in the near future. PMID- 11862672 TI - [Does medicine lose its value near the expiration date?]. AB - This is the question raised to the pharmacists by the patients very often. Medicines, being usually not selected by the customers but prescribed for the patients by their physicians are of a special sort of merchandise. As their efficacy is affected even by psychological factors it is necessary to emphasise that the quality and efficacy of medicines are shown to be constant by the date of expiry. In marketing authorization procedure not only the efficacy and safety but also the quality of the medicines are assessed and a special attention is paid to the chemical, physico-chemical and microbiological stability of the products. Detailed documentation on stability together with the good manufacturing practice of the pharmaceutical factories inspected regularly by the authorities and the system of post marketing surveillance assure that medicines do not loose their good quality and consequently their efficacy and safety by the expiry date stated on the label. PMID- 11862673 TI - [Methods, developments and applications of circular dichroism and simultaneous dual CD/UV spectroscopy in analytical chemistry]. AB - Circular dichroism (CD) is one of the few sensitive indicators of molecular configuration. CD spectroscopy has therefore long been used for structure elucidation. Recent trends show, however, an enormous progress of CD spectroscopy in analytical chemistry, either as an independent method, or as a detector in hyphenated separation techniques. Its uses became especially widespread in pharmaceutical analyses, since enantiomeric purity is a crucial criterion of drug quality. Objectives of the present Ph.D. work included fundamental research in CD and related methodologies, but also, its applications in pharmaceutical analysis. A technical prerequisite of our research work was the instrumental hyphenation of separation and chiroptical detection techniques, which has been achieved by the combination of high performance liquid chromatographic and circular dichroism spectroscopic devices. Our home-built apparatus was the first HPLC-CD instrument in Hungary. Its uses allowed the elaboration of a new method for the determination of peak homogeneity, which applies simultaneous, dual CD/UV detection, and a subsequent ratio-formation. An improved version of this method resulted in the deconvolution of overlapping chromatographic peaks. Another type of peak homogeneity control method was elaborated by rapid, on-line recording of CD spectra in the progress of elution, in order to analyse apparently homogeneous peaks. Circular dichroism has also been applied to quantitate enantiomeric purity in composite samples without any separation of the analytes, and also, to determine equilibrium constants of protonating chiral compounds. PMID- 11862674 TI - [Synthesis of phosphoramide mustard analogues belonging to the L-sugar series]. AB - During the past decades numerous cyclophoshamide (mustard) derivatives of nucleosides and aminodeoxy sugars have been prepared for investigating their antitumor activities. The cyclophosphamide analogues of aminotrideoxy hexoses belonging to the D-series of sugars have been prepared by Monneret et al. The present paper reports the synthesis of the new phosphoramide mustards 16-17 from 12 and 15 (belonging to the L-sugar series). First compound 10 was synthesized from the L-rhamnose (9). Methyl 3-azido-2,3,6,-trideoxy-alpha-L-ribo hexopyranoside (11) was obtained by the replacement of the 3-O-p-toluene-sulfonyl group of 10 with sodium azide. Methyl 3-azido-2,3,6,-trideoxy-alpha-L-arabino hexopyranoside (14) was synthesized by rign opening of 13 with sodium azide. The corresponding amino sugars (12, 15) were obtained by catalytic hydrogenation (over palladium on carbon) of 11 and 14. Our compounds 12 and 15 were transformed into the cyclophosphamide derivatives 16a,b-17a,b upon treatment with bis(2 chloroethyl)phoshoramidic dichloride in the presence of triethylamine (36 h, r.t.). The approximately 1:1 mixtures of isomers (due to the different steric position of the P=O group) could be readily separated by chromatography. The 1H NMR assignments of compounds 16a, 16b, 17a and 17b, were based on one-dimensional selective decoupling experiments or two-dimensional chemical shift-correlated spectroscopy (COSY-60). The assignment of configuration to the isomeric phosphoramidates was based on the magnetic anisotropy of the P=O bond. The distinctly different chemical shift patterns of sugar protons observed for the two isomers allowed the unambiguous assignment of the P=O stereochemistry. The compounds 16a,b-17a,b (mixture of isomers) were tested for inhibitory activity using L1210 and HT29 cell lines. PMID- 11862676 TI - [Torrey Pines Institute for molecular studies]. PMID- 11862675 TI - [The history of heroin]. AB - The discovery of heroin and the development of heroin abuse are introduced. Heroin, the hydrochloride of diacetylmorphine, was discovered by acetylation of morphine. Heroin, in pharmacological studies, proved to be more effective than morphine or codeine. The Bayer Company started the production of heroin in 1898 on a commercial scale. The first clinical results were so promising that heroin was considered a wonder drug. Indeed, heroin was more effective than codeine in respiratory diseases. It has turned out, however, that repeated administration of heroin results in the development of tolerance and the patients become heroin addicts soon. In the early 1910s morphine addicts "discovered" the euphorising properties of heroin and this effect was enhanced by intravenous administration. Heroin became a narcotic drug and its abuse began to spread quickly. Restrictions on its production, use and distribution were regulated by international treties. The total ban on heroin production was also proposed. As a result of the strict regulations the production and cosumption of heroin showed a significant decrease after 1931. At the same time the underworld recognized the shortage of heroin and started the illicit production and trafficking. The quantity of heroin seized by law enforcement agencies in the past decades rose gradually. As an indicator of the worldwide heroin market, the quantity of confiscated heroin underwent a tenfold increase since 1970. The paper surveys the most important heroin producing and trafficking countries. Heroin, prepared in clandestine ("kitchen" or "jungle") laboratories, is diluted ("cut") by every member of the illegal heroin distributing chain, i.e. smugglers, traffickers, dealers and vendors. PMID- 11862677 TI - [Portrait of a scientist. Chemistry and medicine are interdependent. Conversation with Gyual Telegdy of the Hungarian Academy of Sciences. Interview by Bela Noszal]. PMID- 11862678 TI - [Etiologies of cerebral palsy and classical treatment possibilities]. AB - Cerebral palsy is a non-progressive disorder of the developing brain with different etiologies in the pre-, peri- or postnatal period. The most important of these diseases is cystic periventricular leukomalacia (PVL), followed by intra and periventricular hemorrhage, hypoxic-ischemic encephalopathy, vascular disorders, infections or brain malformations. The underlying cause is always a damage of the first motor neuron. Prevalence of cerebral palsy in Europe is 2-3 per 1000 live births with a broad spectrum in different birth weight groups. Our own data concerning only pre-term infants in the NICU with birth weight below 1500 g (VLBW) are between 10%-20%. Established classical treatment methods include physiotherapy (Bobath, Vojta, Hippotherapy), methods of speech and occupational therapists (Castillo-Morales, Sensory Integration) and other therapeutical concepts (Peto, Affolter, Frostig). PMID- 11862679 TI - [Changes in breast feeding behavior in Styria 1994 and 2000]. AB - Aim of this study was to examine breast feeding behaviour in Styria, Austria in the years 1994 and 2000. Differences were documented and possibilities for better support were outlined. 1994 106 and 2000 60 mothers were questioned about breast feeding behaviour of the last child after delivery of a newborn in our hospital. The social status of the parents, breast feeding duration and frequency, problems while breast feeding and contraception were documented. The mothers were also asked about the breast feeding behaviour they planed for the newborn. 1994 83% and 2000 88% of the mothers breast fed their babies for at least 1 month. The average breast feeding duration increased from 4.8 months in 1994 to 5.3 months in 2000. The frequency of breast feeding also increased. Low social status was correlated with shorter breast feeding duration. Support for breast feeding was used by 53% of the mothers in 1994 and by only 28% of the mothers in 2000. 27% of the mothers wish to breast-feed the newborn longer than the last child in 2000. Breast feeding duration has increased in Styria, Austria from 1994 to 2000. Since mothers want to breast feed even longer, support to breast feeding mothers should be offered as often as possible. PMID- 11862680 TI - [Selected blood coagulation problems in newborn infants]. AB - The paper deals with specific features of the neonatal coagulation system. The plasma concentration of the majority of coagulation factors is lower in neonates than in adults, whereby near-adult values are achieved for most components by 6 months of life. Bleeding episodes in haemophilic newborns are rare but may be of a profound nature, thus a proper communication between all involved physicians may aid in the management of such patients. On the other hand thromboembolic episodes might complicate neonatal intensive care, in particular in patients with an indwelling central line. Also inherited thrombophilia might contribute to the development of neonatal thrombosis. Prophylaxis and therapy of vitamin K deficiency bleeding are discussed. PMID- 11862682 TI - ["Neonatology"]. PMID- 11862681 TI - [Cystic periventricular leukencephalomalacia]. AB - Cystic periventricular leukomalacia refers to necrosis of the white matter in a characteristic distribution dorsal and lateral to the external angles of the lateral ventricles in preterm infants. The pathogenesis includes either hypoxic ischaemic lesions resulting from impaired perfusion at the vascular border zones or the role of intra-amniotic infection with toxic effects of endotoxins and cytokines on oligodendrocytes. This overview illustrates the pathogenic theories, risk factors, diagnosis by cranial ultrasonography, and the actual classification. Cystic periventricular leukomalacia is the most severe and frequent cause of cerebral palsy in preterm infants and is almost constantly associated with serious subsequent neuromotor impairments such as diplegia or tetraplegia. Dependant on site and extension of the cysts additionally visual impairments, seizure disorders, hearing impairments, mental retardation, and microcephaly are observed. PMID- 11862683 TI - [Routine examination of the term newborn infant]. AB - The routine examination of healthy term newborn infants on the postnatal ward has at least three goals: diagnosis of a primarily inapparent, yet severe disorder (e.g. acyanotic malformation of the heart), detection of minor, benign abnormalities (e.g. deformed feet) and minor birth lesions (e.g. cephalhematoma, clavicular fracture). Since such findings may cause concern, they are discussed with the parents. Many minor problems such as postural deformities are self limited or can be managed by the mother. A severe disease is detected only rarely. Certain findings should give rise to special investigations (e.g. preauricular tags--urinary tract malformation, hearing impairment). PMID- 11862684 TI - [Virus infections in pregnancy--neonatal management]. AB - We report on the most important viral infections in pregnancy. The importance of these infections is based on severe consequences for foetus, neonate and mother herself. New tests, e.g. direct virus detection assays, improved the security of diagnosis. We emphasize the management of viral infected neonates including own experiences. PMID- 11862685 TI - [Respiratory management of in neonatal intensive care]. AB - This paper gives an critical overview over different therapies for the management of neonates with respiratory failure. Several, partially revolutionary, new therapies have been introduced in the clinical routine in the last 10 to 15 years like surfactant therapy, inhalation of nitric oxide, extracorporeal membrane oxygenation and high frequency oscillatory ventilation. Furthermore, there are new promising ventilatory techniques like pressure support ventilation, proportional assist ventilation or liquid ventilation in clinical testing. PMID- 11862686 TI - ["100 rats and 20 children!"--Julius Moses and the debate over human experiments in the Weimar Republic]. AB - In 1900, the Prussian ministry of cultural affairs for the first time issued ethical guidelines on human experimentation. These guidelines were not able to prevent questionable human experiments in Germany during the Weimar Republic. In 1928 the testing of a vitamin-D-medication in the Berlin "Kaiserin Auguste Victoria-Haus" led to a public scandal. The physician and active social democrat Julius Moses played a decisive role in the disclosure of this scandal and of other inadmissible experiments. Subsequently in 1930, the "Reichsgesundheitsrat" developed improved guidelines distinguishing between scientific and therapeutic experiments. PMID- 11862687 TI - [Results of mechanical ventilation in premature infants with respiratory distress syndrome]. AB - This paper reveals the results of synchronized intermittent mandatory ventilation in premature babies over a period of 8 years. Out of 1532 mechanical ventilated newborns 438 were prematures with severe hyaline membrane disease. The incidence of barotraumatic complications (Pneumothorax 1.2%, pulmonary interstitial emphysema 3% and bronchopulmonary dysplasia 3.4%) was relatively low. We discuss this as an effect of using low respirator frequencies guaranteering sufficient emptying time of the lungs. PMID- 11862688 TI - [Cerebral hemorrhage in term newborn infants--an analysis of 10 years (1989 1999)]. AB - About 1 out of 100 full term born infants develops intracranial hemorrhage. The study represents an analysis of intracranial hemorrhage over a ten-years period. Diagnosis was based on ultrasound scan. A division into five major bleeding types was done according to the location of the hemorrhage. The neurological outcome was reviewed. Patients were classified into three groups according to their neurological outcome: normal neurological development, mild neurological impairment, and severe neurological impairment. Between 1989-1999, 2019 full-term newborn infants were treated at the neonatal intensive care unit in Graz. 54 of them (2.7%) developed intracranial hemorrhage, of whom 13 (24%) suffered from subdural hemorrhage, 1 (2%) from primary subarachnoid hemorrhage, 14 (26%) from hemorrhage within the choroid plexus, 6 (11%) from intraventricular hemorrhage grade I (IVH I), 11 (20%) from intra- and periventricular hemorrhage (IVH III/PVH), 6 (11%) from hemorrhage within the basal ganglia, and 3 (6%) from intracerebellar hemorrhage. Considering the neurological outcome, 34 (63%) were diagnosed as being developed normally, 15 (28%) developed any neurological impairment, 3 (5%) died, whereas 2 (4%) did not participate our follow up program. PMID- 11862689 TI - The scientific evaluation of mental health treatments: an historical perspective. PMID- 11862690 TI - Evidence-based practice and clinical social work. PMID- 11862691 TI - [Estimation of the developmental stability of small-leafed lime on natural reserves and in urbanized territories]. AB - The developmental stability of small-leaved lime (Tina cordata Mill.) was estimated in populations on territories of the Kerzhenskii State Nature Reserve, Nizhni Novgorod District, and in an industrial region of Nizhni Novgorod. The developmental stability was estimated according to the fluctuating asymmetry of the leaf measurements. The results obtained suggest that, in the urbanized territory, the homeostatic developmental mechanisms are weakened, as expressed in the increased degree of leaf asymmetry. PMID- 11862692 TI - [Invagination and evagination: a hydromechanical model]. AB - It has been proposed that processes of invagination and evagination be considered in the context of a single, "hydromechanical," model. The model is based on the assumption that the structures demonstrating invaginations and evaginations are closed systems capable of changing their intracavitary pressure in an autonomous regime. The mass, which occupies the cavity, should overcome the resistance of the surrounding membrane during its growth. In places of the weakest resistance, the inner mass expands especially quickly: it bulges out. If the pressure under the membrane is less than the exterior one, the process goes in the opposite direction: the outer mass intrudes (invaginates) into the cavity, causing its wall to sag in the weakest regions. PMID- 11862693 TI - [A study of hemopoiesis on a sublayer of peritoneal cells in the presence of hemopoietic cytokines]. AB - We studied the effects of erythropoietin and thrombopoietin on the clonogenic capacity and direction of differentiation of the hemopoietic cells that form colonies on acetate cellulose membrane in the peritoneal cavity of mice. An increased level of erythropoietin in the blood of recipient mice after blood letting led to the appearance of erythroid colonies upon transplantation of syngeneic hemopoietic cells but did not affect the differentiation of transplanted xenogeneic (guinea pig) hemopoietic cells. Erythropoietin transported top the stromal sublayer by a polymeric carrier also induced erythroid differentiation, while thrombopoietin transported in a similar way somewhat enhanced megakaryocytopoiesis. PMID- 11862694 TI - [Appearance and differentiation of NADPH-D-neurons in ontogeny of the cerebral cortex in rats]. AB - The appearance of presumptive NO-ergic nerve cells and their differentiation in the rat neocortex were studied. For this purpose, a comparative analysis of the development and differentiation of NADPH-D-positive neurons in the neocortex transplants taken from the embryos of different ages and transplanted in the occipital cortex of adult rats and in the normally developing cerebral cortex. The nervous tissue was stained histochemically for NADPH-D. The results we obtained suggest that no NADPH-D-containing neurons were found in the transplants from 15-day embryos, while they developed in those from 18-day embryos. Hence, precursors of NO-ergic neurons were still absent in the presumptive neocortex of 15-day embryos and appeared only on day 16-18 of embryogenesis. Expression of NADPH-D begins in them only within four to five days, but the neurons are differentiated during a relatively short period of time. Most NADPH-D-positive neurons reach their structural-functional maturity already by the end of the first week of postnatal development, while their complete maturation takes place by the end of the second week of postnatal development. PMID- 11862695 TI - [Involvement of accessory neurosecretory nuclei of hypothalamus in the formation of hypothalamohypophyseal system during prenatal and postnatal development in rats]. AB - We studied the development of direct axonal connections of the accessory neurosecretory hypothalamic nuclei with the posterior pituitary lobe on the fixed rat brain from day 15 of embryogenesis until day 10 of postnatal development using the retrograde diffusion method of the lipophilic fluorescent carbocyanine dye 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate along the neuronal membranes. The marker was applied onto the posterior pituitary lobe and, after incubation in a fixative, fluorescing bodies of nerve cells were visualized in the hypothalamus. Neuronal axons of the retrochiasmatic nucleus were the first of the accessory nuclei to ingrow in the posterior pituitary lobe (on days 16-17 of embryogenesis). Neurons of the circular and dorsolateral nuclei and the nuclei of the median bundle of the forebrain sent their axons to the posterior pituitary lobe starting from the first days of postnatal development. No direct connections of the anterior commissure and perifornical accessory nuclei with the posterior pituitary lobe were found in perinatal development. These facts are discussed in the light of concepts about the different functional role of accessory peptidergic hypothalamic nuclei in rats. PMID- 11862696 TI - [Ontogeny of different life forms of plants and specific features of age and spatial structure of their populations]. AB - Materials have been generalized that were accumulated in population-ontogenetic studies of plants and leading features of the ontogeny of plants have been noted that determine specific structural features of their populations. The described patterns allowed the authors to develop simulation models of plant population dynamics. The results of simulation are provided, and they are compared to the empirical data. Problems have been considered that concern the influence of specific features of development of different plant biomorphs on the genetic structure of populations and biocoenotic processes. PMID- 11862698 TI - [Effects of growth factors FGF2 and IGF2 on the development of parthenogenetic mouse embryos in utero and in vitro]. AB - We studied the effects of fibroblast growth factor 2 (FGF2) and insulin-like growth factor 2 (IGF2) on the development of parthenogenetic mouse embryos (CBA x C57BK/6)F1. The parthenogenetic embryos were treated in vitro during the preimplantation period and, at the blastocyst stage, transplanted into the uterus of pseudopregnant females. The addition of FGF2 at an optimal dose (2.5 ng/ml) to the culture medium increased twofold the number of embryos developed in utero to the somite stages as compared to the control: 18 and 43%, respectively. The parthenogenetic embryos (18-21 somites), treated and nontreated with FGF2 during the preimplantation period, were explanted for further development in vitro and treated with IGF2 at 2.5 micrograms/ml. As a result, many more parthenogenetic embryos (> 87%) of both groups developed in vitro to the stage of 30 or more somites as compared to the control (59%). The treatment of the parthenogenetic embryos with FGF2 alone at the preimplantation stages did not improve their development in vitro at the postimplantation stages. The results we obtained suggest that the treatment of parthenogenetic embryos in vitro with FGF2 during the preimplantation period increased twofold the number of somite embryos in utero, while their subsequent treatment in vitro with IGF2 leads to a significant prolongation of their development, as compared to the control. PMID- 11862697 TI - [Cytoskeletal control of cell length regulation]. AB - It was shown that mouse embryo fibroblasts and human foreskin diploid fibroblasts of AGO 1523 line cultivated on specially prepared substrates with narrow (15 +/- 3 microns) linear adhesive strips were elongated and oriented along the strips, but the mean lengths of the fibroblasts of each type on the strips differed from those on the standard culture substrates. In contrast to the normal fibroblasts, the length of mouse embryonic fibroblasts with inactivated gene-suppresser Rb responsible for negative control of cell proliferation (MEF Rb-/-), ras transformed mouse embryonic fibroblasts (MEF Rb-/-ras), or normal rat epitheliocytes of IAR2 line significantly exceeded those of the same cells on the standard culture substrates. The results of experiments with the drugs specifically affecting the cytoskeleton (colcemid and cytochalasin D) suggest that the constant mean length of normal fibroblasts is controlled by a dynamic equilibrium between two forces: centripetal tension of contractile actin-myosin microfilaments and centrifugal force generated by growing microtubules. This cytoskeletal mechanism is disturbed in MEF Rb-/- or MEF Rb-/-ras, probably, because of an impaired actin cytoskeleton and also in IAR2 epitheliocytes due to the different organization of the actin-myosin system in these cells, as compared to that in the fibroblasts. PMID- 11862699 TI - [Specific alterations in the secondary structure of eukaryotic DNA effected by a tetrahydrocortisol-apolipoprotein A-I complex]. PMID- 11862700 TI - [Replication protein RepN encoded by the RC plasmid of thermophilic bacterium Thermoanaerobacterium saccharolyticum: mutational analysis and deletion mapping of domains responsible for its lethal effect]. AB - Amino acid sequence analysis of the product encoded by repN of Thermoanaerobacterium saccharolyticum (Clostridium thermosaccharolyticum) pNB2, which is capable of rolling-circle (RC) replication, revealed all known motifs conserved among replication (Rep) proteins that initiate RC replication of plasmids related to pC194/pUB110. Using the T7 expression system in Escherichia coli, RepN was identified as a 35K protein. Its lethal effect on bacterial cells was unusually high for a protein of the kind. Mutation analysis of the potential active centers (Y85F and Y211F) showed that the lethal effect of RepN is not associated with its putative topoisomerase (relaxase) activity. On evidence of deletion mapping, the lethal effect was attributed to the N- and C-terminal domains, each accounting for about 30% of the total protein. The RepN fragments essential for the lethal effect were found to share a motif, which showed no appreciable homology to known conserved motifs. The high lethal effect of RepN was assumed to result from duplication of the motif and to play an adaptive role, providing for the stable maintenance of the AT-rich plasmid in thermophilic bacterial cells. PMID- 11862701 TI - [Crosslinking of mRNA analog, pUUAGUAUUUAUU derivative with a photoactivated group at the guanosine residue, with human 80S ribosomes]. AB - Crosslinking of mRNA analog, dodecaribonucleotide pUUAGUAUUUAUU derivative carrying a perfluoroarylazido group at the guanine N7, was studied in model complexes with 80S ribosomes involving tRNA and in binary complex (i.e., in the absence of tRNA). It was shown that, irrespectively of complex formation conditions (13 mM Mg2+, or 4 mM Mg2+ in the presence of polyamines), the mRNA analog in binary complex with 80S ribosomes was crosslinked with sequence 1840 1849 of 18S rRNA, but in the complexes formed with participation of Phe-TPHKPhe (where the G residue carrying the arylazido group occupied position-3 to the first nucleotide of the UUU codon at the P site) the analog was crosslinked with nucleotide 1207. The presence and the nature of tRNA at the E site had no effect on the environment of position-3 of the mRNA analog. Efficient crosslinking of the mRNA analog with tRNA was observed in all studied types of complex. Modified codon GUA, when located at the E site, underwent crosslinking with both cognate valine tRNA and noncognate aspartate tRNA for which the extent of binding at the E site of 80S ribosomes was almost the same and depended little on Mg2+ concentration and the presence of polyamines. PMID- 11862702 TI - [In vitro formation of triple-stranded DNA structures in the human alpha1 antitrypsin gene]. AB - We analyzed the possibility of triplex formation in the human alpha 1-antitrypsin gene. Conditions and nucleotide sequence dependence of triplex formation and thermostability of the product were determined. PMID- 11862703 TI - [A new method to measure the functional activity of class-1 translation termination factor eRF1]. AB - Termination of protein synthesis (hydrolysis of the last peptidyl-tRNA on the ribosome) takes place when the ribosomal A site is occupied simultaneously by one of the three stop codons and by a class-1 translation termination factor. The existing procedures to measure the functional activity of this factor both in vitro and in vivo have serious drawbacks, the main of which are artificial conditions for in vitro assays, far from those in the cell, and indirect evaluation of activity in in vivo systems. A simple reliable and sensitive system to measure the functional activity of class-1 translation termination factors could considerably expedite the study of the terminal steps of protein synthesis, at present remaining poorly known, especially in eukaryotes. We suggest a novel system to test the functional activity in vitro using native functionally active mRNA, rather than tri-, tetra-, or oligonucleotides as before. This mRNA is specially designed to contain one of the three terminating (stop) codons within the coding nucleotide sequence. Plasmids have been generated that carry the genes of suppressor tRNAs each of which is specific toward one of the three stop codons. They were shown to support normal synthesis of a reporter protein, luciferase, by reading through the stop codon within the coding mRNA sequence. We have demonstrated that human class-1 translation termination factor eRF1 is able to compete with suppressor tRNA for a stop codon and to completely prevent its suppressive effect at a sufficient concentration. Forms of eRF1 with point mutations in functionally essential regions have lower competitive ability, demonstrating the sensitivity of the method to the eRF1 structure. The enzymatic reaction catalyzed by the full-size reporter protein is accompanied by emission of light quanta. Therefore, competition between suppressor tRNA and eRF1 can be measured using a luminometer, and this allows precise kinetic measurements in a continuous automatic mode. PMID- 11862704 TI - [Comparative study of the M.Bstf5I-1 and M.BstF5I-3 DNA methyltransferases from the Bacillus stearothermophilus F5 restriction-modification system]. AB - The BstF5I restriction-modification system from Bacillus stearothermophilus F5, unlike all known restriction-modification systems, contains three genes encoding DNA methyltransferases. In addition to revealing two DNA methylases responsible for modification of adenine in different DNA strands, it has been first shown that one bacterial cell has two DNA methylases, M.BstF5I-1 and M.BstF5I-3, with similar substrate specificity. The boundaries of the gene for DNA methyltransferase M.BstF5I-1 have been verified. The bstF5IM-1 gene was cloned in pJW and expressed in Escherichia coli. Homogeneous samples of M.BstF5I-1 and M.BstF5I-3 were obtained by chromatography with different sorbents. The main kinetic parameters have been determined for M.BstF5I-1 and M.BstF5I-3, both modifying adenine in the recognition site 5'-GGATG-3'. PMID- 11862705 TI - [Systematic analysis of buried polar side chains and their interactions in alpha helical proteins]. AB - Examination of 80 alpha-helical proteins and domains demonstrates that they contain from 1 to more than 20 completely buried (water-inaccessible) polar side chains. As a rule the latter have partners for H-bonding but the resulting H-bond system is often not exhaustive. Basing on statistical analysis, we determined the optimal number of H-bonds for every type of polar side chain, and discuss the structural role of vacant donors and acceptors. About half of the H-bonds formed by buried side chains pertain to interhelix contacts of the (side chain)-(side chain) and (side chain)-(main chain) types. Such interactions appear to be a most important factor determining the mutual arrangement of alpha-helices in proteins. Analysis of the frequency of occurrence of various interacting pairs reveals that these interactions are selective. PMID- 11862706 TI - [Enhancing the stability of oligonucleotide duplexes. The effect of adding 1-(2' deoxy-beta-d-ribofuranosyl)-5-nitroindole]. AB - We studied the properties of DNA duplexes containing 5-nitroindole (N) in one of the chains. We synthesized 8-membered oligos with N at the 5' or at the 3' end: 5'-d(NXGACCGTC)-3' or 5'-d(GACCGTCXN)-3', where X is one of the four natural bases, making all four kinds of oligos with and without N. We also prepared 11 membered oligos complementary to the above octanucleotides: 5'-d(TGACGGTCYZT)-3' and 5'-d(TZYGACGGTCT)-3', where Y and Z are A, G, C, or T. The stability of duplexes obtained with these oligos was assessed by melting, and the thermodynamic parameters delta H, delta S, and Tm were calculated. Comparison of the melting curves for modified and nonmodified duplexes demonstrated that the presence of N at the 5' end of one chain raises the Tm by 6.6 degrees C on average; if N is at the 3' end of the same chain, the Tm increases by about 3 degrees C. PMID- 11862708 TI - [Streptomyces rimosus aminoglycoside-3'-phosphotransferase VIII: comparisons with aminoglycoside-3'-phosphotransferases of aminoglycoside-producing strains with eukaryotic protein kinases]. AB - The nucleotide sequence was established for the aphVIII aminoglycoside phosphotransferase gene of an oxytetracycline-producing Streptomyces rimosus strain. The gene is 804 bp in size and possibly codes for APHVIII of 267 residues. Heterologous expression of aphVIII was studied in Escherichia coli and Chlamydomonas reinhardtii. The deduced APHVIII sequence was compared with known sequences of aminoglycoside phosphotransferases of aminoglycoside-producing actinomycete strains and of eukaryotic protein kinases. A local homology of 38 residues was found between APHVIII and actinomycete serine-threonine protein kinases in the conserved region possibly involved in ATP binding. APHVIII differed from aminoglycoside 3'-phosphotransferases of aminoglycoside-producing actinomycete strains and of clinical isolates, and can be classed to a separate group. PMID- 11862709 TI - [Structure and function of eukaryotic nuclear DNA-dependent RNA polymerase I]. AB - In the eukaryotic cell, normal protein biosynthesis is sustained by several million ribosomes, which contain rRNA as an essential component. The high molecular-weight precursor of large and 5.8S rRNAs is synthesized by DNA dependent RNA polymerase I (Pol I) in the nucleolus. Data on DNA regulatory elements, protein factors involved in rDNA transcription by Pol I, subunit composition of Pol I, and on the interactions and possible functions of individual subunits are summarized. PMID- 11862707 TI - [Pyrimidine oligodeoxyxylonucleotides form triplexes with purine DNA in neutral media]. AB - Interactions of oligonucleotides comprising (1-beta-D-2'-deoxy-threo pentafuranosyl)thymine and (1-beta-D-2'-deoxy-threo-pentafuranosyl)cytosine residues (oligodeoxyxylonucleotides or OXNs) with complementary single-stranded DNA fragments were investigated. Using nondenaturing gel electrophoresis, footprinting, and melting assays, pyrimidine OXNs were shown to form triplexes with the purine DNA template, which are stable at neutral pH and comparable in heat stability with the corresponding natural polypurine-polypyrimidine DNA duplexes. In such triplexes, the N3 of cytosines in one of the OXNs are protonated. As revealed by CD spectroscopy in the 210-340 nm range, the form of the triple helix depends on the nucleotide composition and sequence of the DNA template, and is intermediate between A and B. PMID- 11862711 TI - [Homologous and heterologous type 2 casein kinases have the same effect on the affinity for RNA of the Gag structural glycoprotein of gypsy (MDG4)]. PMID- 11862712 TI - [Suppression of nonsense mutations in the Dystrophin gene by a suppressor tRNA gene]. AB - Nonsense mutations in the dystrophin gene are the cause of Duchenne muscular dystrophy (DMD) in 10-15% of patients. In such an event, one approach to gene therapy for DMD is the use of suppressor tRNAs to overcome the premature termination of translation of the mutant mRNA. We have carried out cotransfection of the HeLa cell culture with constructs containing a suptRNA gene (pcDNA3suptRNA) and a marker LacZ gene (pNTLacZhis) using their polymer VSST-525 complexes. It was found that the number of cells producing beta-galactosidase depends inversely on the dose of the suptRNA gene. A single in vivo injection of the construct providing for expression of the suptRNAochre gene into mdx mouse muscle resulted in the production of dystrophin in 2.5% of fibers. This suggests that suppressor tRNAs are applicable in gene therapy for hereditary diseases caused by nonsense mutations. PMID- 11862713 TI - [Prediction of recognition sites for genomic replication of classical swine fever virus with information analysis]. AB - In order to explore the mechanism for the genomic replication of classical swine fever virus (CSFV), so as to make a basis for investigating its pathogenicity, an introduction of the information theory is presented in connection with the statistical mechanics, whence small-sample statistics appears naturally as a consequence of the Bayesian approach. Furthermore, a selection rule for identifying the pattern of a recognition site for an RNA-binding protein is proposed by means of the maximum entropy principle. Based on those, the information contents of 3'-untranslated regions (3'UTRs) of genomes of 20 CSFV strains and 5'-untranslated regions (5'UTRs) of genomes of 58 CSFV strains are analyzed with a computational algorithm in a reduction mode, and the 3'UTR sites of 20 strains and 5'UTR sites of 58 strains containing important motifs are extracted from the unaligned RNA sequences of unequal lengths. These sites, which have the patterns of sequence and structure similar to the putative cis elements related to the regulation of genomic replication, would be identified as the potential recognition sites in 3'UTRs and 5'UTRs for CSFV replicase responsible for classical swine fever virus genomic replication, and to some extent, this identification is supported by experimental evidence. Finally, information analysis allows a presumption to be made about the CSFV RNA replication initiation mechanism. PMID- 11862710 TI - [Association of polymorphic trinucleotide repeat (GAA)n of the Frataxin gene with diabetes mellitus type 2 in the Moscow population]. PMID- 11862714 TI - [Transfection with the E1A and E1B-19kDa oncogenes does not prevent rat embryo fibroblasts from cell cycle arrest after gamma-radiation]. AB - Introduction of the E1A early region of the human adenovirus type 5 impairs the ability of mammalian cells to arrest the cell cycle at G1/S after damage. Two parameter fluorescent-activated cell sorting (FACS) with iododeoxyuridine revealed the radiation-induced G1/S arrest in rat embryo fibroblasts transformed with the complementing E1A + E1B-19 kDa oncogenes. This was due to selective inhibition of CycIE/Cdk2-associated kinase activity, while activities of type 2 kinase and of CyclA/Cdk2 complexes remained unchanged. The inhibitor of G1-phase cyclin kinases, p21/Waf1, was accumulated and interacted with target kinases both in normal and in transformed cells after irradiation. As shown by immunoprecipitation, p21/Waf1 formed complexes with the E1A on coproducts in the transformants, which possibly accounted for its functional inactivation. Kinase modification in cyclin-kinase complexes was assumed to play a key role in regulation of cyclin-dependent kinases in the transformants with inactivated p21/Waf1. PMID- 11862716 TI - [A short form of the heparin-binding EGF-like growth factor with a changed EGF domain]. AB - In all secreted proteins related to the epidermal growth factor (EGF), EGF domains that occur in a mature factor are each encoded by two exons, and those that do not, by one exon. During splicing, additional exon 3a can be inserted between exons 3 and 4, which code for the EGF domain of the mature heparin binding EGF-like growth factor (HB-EGF). The resulting mRNA codes for the short form of HB-EGF (SF HB-EGF), which retains the signal peptide, the propeptide, and the heparin-binding domain. However, its EGF domain lacks the C-terminal subdomain essential for the interaction with the EGF receptor (EGFR). Structural analysis suggested that SF HB-EGF is a secreted polypeptide that has high affinity for heparin, but weakly, if at all, interacts with EGFR. Data obtained in three different systems indicated that SF HB-EGF possesses a mitogenic activity but utilizes a signal transduction pathway other than that of HB-EGF. PMID- 11862717 TI - [The immune response to artificial proteins containing biologically active fragments of human IFN-alpha2 and insulin]. AB - A study was made of the humoral immune response of BALB/c mice to various doses of artificial proteins that contained biologically active fragments of human interferon alpha 2 (IFN-alpha 2) and insulin. The insulin fragment had no effect on the response to any protein construct. The IFN-alpha 2 fragment increased the titer of antibodies against the construct. Mapping of continuous B epitopes with immune sera revealed several antigenic determinants, the C end of the IFN-alpha 2 fragment with the adjacent de novo protein region being immunodominant. More effective binding of serum antibodies with the constructs containing the IFN alpha 2 fragment was attributed to antibody interaction with the fragment and to a better recognition of the entire protein construct by the immune system. PMID- 11862718 TI - [The ratio of prolactin receptor isoforms in rat hepatocytes: the effect of obstructive cholestasis]. PMID- 11862719 TI - [Immortalization of human cells and induction of dna synthesis in heterokaryons]. PMID- 11862715 TI - [Role of the TCF phosphorylation state and the chromatin structure in the negative transcription regulation of the c-fos proto-oncogene in E1A + c-Ha-ras transformed cells]. AB - As compared with normal rat embryo fibroblasts (REF), c-fos transcription is suppressed in REF transformed with the E1A and cHa-ras complementing oncogenes. Negative regulation of the fos promoter is due to the serum-responsive element (SRE) constantly bound with the serum response factor (SRF) and with the ternary complex factor (TCF), which form the SRF/SRF/TCF complex. Possible mechanisms of the c-fos suppression were studied, including changes in the TCF content and phosphorylation and replacement of TCF by repressor proteins. In addition, the transcription factors were tested for DNA-binding activity, and c-fos transcription was analyzed by RT-PCR in various cell culture conditions and in the presence of phosphatase and histone deacetylase inhibitors. The c-fos suppression in transformed REF was explained by formation of a transcriptionally inactive chromatin structure in the fos promoter region as a result of histone deacetylation, rather than by insufficient TCF phosphorylation. PMID- 11862721 TI - Recurrent pneumonia in children. PMID- 11862720 TI - [Interaction of tetrahydrocortisol-apolipoprotein A-I complex with eukaryotic DNA and with single-stranded oligonucleotides]. AB - The complex formed by tetrahydrocortisol (THC) and apolipoprotein A-I (ApoAI) specifically interacts with eukaryotic DNA from rat liver. Taken together, physical and chemical data and the results of small-angle X-ray scattering analysis show that interaction of the THC-ApoAI complex with eukaryotic DNA results in deformation of the DNA double helix. Single-stranded fragments were demonstrated to cause deformation of the double helix. In this state DNA forms complexes with DNA-dependent RNA polymerase. This interaction is cooperative and of saturating type; up to six enzyme molecules bind with one DNA molecule. The putative site of complex binding with DNA is the sequence CC(GCC)n found in many genes including the human ApoAI gene. An oligonucleotide of this type was synthesized. Its association constant (Ka) was 1.66 x 10(6) M-1. Substitution of THC with cortysol considerably decreases the Ka. We suggest that THC interacting with GC pairs of the binding site forms hydrogen bonds with cytosine, inducing rupture of the bonds within the complementary nucleic base pair. PMID- 11862722 TI - Respiratory tract infections in cystic fibrosis. PMID- 11862723 TI - Croup. PMID- 11862724 TI - Respiratory infections in children in developing countries. PMID- 11862725 TI - Resident's column: what every resident should know about tuberculosis. PMID- 11862726 TI - A pediatrician's view. Pulmonary infection still happens. PMID- 11862728 TI - Bronchiolitis. PMID- 11862730 TI - HAART patients can face greater risk of developing heart disease--studies. AB - Investigators from the HIV Outpatient Study group, which is located at 10 sites in the United States, found that after a few years of taking protease inhibitors, HIV patients were beginning to show a significantly increased rate of myocardial infarctions. The researchers presented their findings at the 2001 IDSA Conference, held Oct. 25-28, 2001. PMID- 11862729 TI - Pediatric tuberculosis. PMID- 11862731 TI - Nucleic acid test system approval. PMID- 11862732 TI - Tenofovir (viread) receives FDA approval. PMID- 11862733 TI - Truegene HIV-1 genotyping kit approval. PMID- 11862734 TI - Easier abacavir regimen has promising results. AB - HIV patients, understandably, are sometimes unenthusiastic about taking their protease inhibitor (PI) drug therapies because of the numerous pills, the side effects, and chronic problems such as lipodystrophy. To address these problems, researchers have been studying non-PI drug therapies that are potent against HIV. Research has demonstrated successful viral suppression for at least 24 weeks when patients were switched from a PI regimen to an abacavir regimen. PMID- 11862735 TI - Opportunistic infection improvements level off. AB - One of the greatest achievements of protease inhibitor and highly active antiretroviral therapy has been the dramatic decrease in the incidence of opportunistic infections (OIs) related to AIDS. In the first couple of years after the advent of PI therapy, clinicians found that fewer HIV/AIDS patients developed some of the more common OIs, such as Kaposi's sarcoma and pneumocystosis. However, new research shows that the decline in OI rates has ended. PMID- 11862736 TI - New research adds support for ED at-risk testing. AB - A study conducted at the University Hospitals of Cleveland in Ohio adds to the growing body of research that suggests it's a good strategy to offer HIV testing to patients admitted to hospital emergency departments. Offering testing to emergency room patients reaches a different group that's at risk, such as African American women, says one researcher. PMID- 11862737 TI - Infection with GBV-C linked to longer life. AB - New research has established a link between HIV patients being infected with a virus that is similar to hepatitis C and having a slower HIV disease progression. Investigators first began to study GBV-C in the mid-1990s as a possible cause of chronic hepatitis, but they quickly lost interest when the virus was shown to have no direct link to the disease. PMID- 11862738 TI - CDC posts new HIV testing, referral guidelines. AB - New HIV testing guidelines released by the Centers for Disease Control and Prevention include strategies for increasing early testing of at-risk people and universal HIV testing of pregnant women. The guidelines describe state-of-the-art HIV testing, counseling, and referral services. PMID- 11862739 TI - Evidence shows hepatitis C virus is playing major role in AIDS deaths. AB - HIV patients who are coinfected with hepatitis C may be at a greater risk of dying from their hepatitis infection than from untreated HIV, according to research presented at the 41st Interscience Conference on Antimicrobial Agents and Chemotherapy. Experts suggested that when an HIV patient's CD4 cell count is greater than 350, it may be a better strategy to first treat the patient for hepatitis C for up to six months and then begin antiretroviral treatment if indicated. PMID- 11862740 TI - Drug resistance test shows encouraging results. AB - Researchers continue to find evidence that the combination of lopinavir/ritonavir (Kaletra) provides prolonged potency while avoiding drug resistance. The Kaletra findings come on the heels of another study that found a high rate of drug resistance among HIV patients with measurable levels of virus in their blood. PMID- 11862741 TI - Tests on new NNRTI show strong potency. AB - Early clinical studies of TMC125, a new non-nucleoside reverse transcriptase inhibitor, indicate the drug is extremely potent, with an average reduction in HIV viral load of 99% after one week of treatment. PMID- 11862742 TI - Fears of an Eastern European explosion of HIV epidemic are being realized. AB - The number of people living with HIV in Eastern Europe doubled last year, fueled primarily by high unemployment, economic insecurity, liberal social and cultural norms, and disintegrating public health services, according to UNAIDS. PMID- 11862743 TI - Sub-Saharan strategies best when tailor-made. PMID- 11862744 TI - Consider postexposure STD screening. PMID- 11862745 TI - One copy of mutation may help resistance: study. AB - It's well-known that the homozygous type of two copies of the 32 base pair deletion in the CCR5 chemokine receptor provides resistance to HIV-1 infection. But there have been fewer data pertaining to what happens when a person has only one copy of the CCR5 delta 32 genotype. A recent study shows that even heterozygous CCR5 32 genotype, found primarily in European Caucasians, provides significant protection from infection during exposure to HIV-1. PMID- 11862746 TI - HIV/Ebola comparison could spur new treatments. AB - A researcher has discovered a link between HIV and Ebola virus: Both viruses use the same method to spread through the human body. PMID- 11862747 TI - NIH studies link between saquinavir and garlic pills. PMID- 11862748 TI - UCSF launches web site to educate youths. PMID- 11862749 TI - Expert offers help for PI-related heart disease. AB - Joseph J. Eron Jr., MD, director of Clinical Core at the University of North Carolina Center for AIDS Research in Chapel Hill, discusses the recent findings that HAART and protease inhibitors may increase risk of heart disease in HIV patients. Eron also offers a strategy for treating patients at risk for heart disease. PMID- 11862750 TI - FDA notifications NucliSens HIV-1 QT is approved. PMID- 11862751 TI - New director appointed for FDA's DAVDP unit. PMID- 11862752 TI - [Role of nitric oxide in survival and death of neurons]. AB - The prominent pathological feature of the brain in Parkinson's disease is selective degeneration of dopaminergic neurons in the substantia nigra of the midbrain. Glutamate and nitric oxide (NO) are the major effectors of the radical stress that may induce selective loss of dopaminergic neurons. It has been postulated that neurotoxicity induced by glutamate and NO in dopaminergic neurons is regulated by certain endogenous factors. We have reported that estradiol protects dopaminergic neurons against NO-mediated glutamate neurotoxicity by reducing intracellular reactive oxygen species (ROS) levels. We further searched for a candidate for neuroprotective substances with unique structure. From the ether extract of fetal calf serum (FCS), we isolated a novel substance possessing protective activity against neurotoxicity induced by glutamate NO. The compound was a sulfur-containing diterpenoid and showed hydroxyl radical scavenging activity. We further analyzed the change of resistance to excitotoxicity in midbrain dopaminergic neurons in co-culture with the striatum by using a slice culture technique. The results suggested that the generation of NO is involved in NMDA cytotoxicity on dopaminergic neurons and that increased activity of SOD in co-culture renders dopaminergic neurons resistant to NMDA cytotoxicity by preventing peroxynitrite formation. Those findings suggest that regulation of intracellular ROS levels plays a critical role in protecting neurons against NO mediated radical stress in neurodegenerative disorders. PMID- 11862753 TI - [Nitroxidergic (nitrergic) nerve and erectile dysfunction]. AB - In vascular tissues including the corpus cavernosum, the organ function is reciprocally regulated by noradrenergic and non-adrenergic, non-cholinergic (NANC) nerves. NANC nerves innervating the corpus cavernosum is thought to be nitroxidergic (nitrergic) nerves which liberate nitric oxide (NO) produced by neuronal NO synthase, and liberated NO activates soluble guanylate cyclase (sGC) in cavernous smooth muscle cells. Intracellular increase in cyclic (c) GMP by activation of sGC dilates cavernous smooth muscle and then induces penile erection. Nitroxidergic (nitrergic) vasodilator nerves also innervate cavernous arteries and veins which regulate the blood volume in the corpus cavernosum. The order of potency of nitroxidergic nerve functions in these tissues (cavernosum > artery >> vein) may be suitable for producing the erection. Therefore, obstruction of the arteries and impairment of nitroxidergic (nitrergic) nerve function are speculated to be one of the causes for erectile dysfunction (ED). On the other hand, NO derived from the cavernous endothelium may partly contribute to erectile function. Sildenafil (Viagra) is one of the potent therapeutics for ED. The agent is a selective phosphodiesterase type 5 (PDE-V) inhibitor that inhibits degradation of cGMP elevated by NO mainly derived from the nerves. To develop more selective and safer therapeutics for ED, further systematic investigations are required. PMID- 11862754 TI - [Biological and pathophysiological roles of endogenous methylarginines as inhibitors of nitric oxide synthase]. AB - Protein arginine N-methyltransferases (PRMTs) catalyse the methylation of guanidinonitrogen(s) of arginine to produce NG-monomethyl-L-arginine (L-NMMA), asymmetric NG,NG-dimethyl-L-arginine (ADMA) and symmetric NG,NG-dimethyl-L arginine (SDMA), which are subsequently released into the cytoplasm following proteolysis. Free intracellular L-NMMA and ADMA, but not SDMA, are inhibitors of all three isoforms of nitric oxide synthases (nNOS, eNOS and iNOS). L-NMMA and ADMA, but not SDMA, are actively metabolized by dimethylarginine dimethylaminohydrolase (DDAH) to L-citrulline and methylamine (and dimethylamine). Free methylarginines are detectable in cell cytosol, plasma and tissues. Elevated ADMA has been detected in the plasma of patients or experimental animals with hypercholesterolemia, renal failure, atherosclerosis, hypertension, thrombotic microangiopathy, peripheral arterial occlusive disease and in the regenerated endothelial cells after angioplasty. Moreover, in the non cardiovascular field, ADMA was increased in the urethral tissue following ischemia and in the plasma of patients with schizophrenia and multiple sclerosis. Altered biosynthesis of NO has been implicated in the pathogenesis of these diseases, and it is possible to consider that the accumulation of endogenous L NMMA and ADMA underlies the impaired NO generation and increased O2- production. We described herein the biosynthesis, transmembrane transport, metabolic pathway and possible pathophysiological roles of endogenous methylarginines. PMID- 11862755 TI - [Somatic gene therapy of dilated cardiomyopathy]. AB - The hereditary form of dilated cardiomyopathy (DCM) accounts for about 20% of human DCM and is a major cause of heart failure. TO-2 strain hamsters show DCM, a gene deletion of delta-sarcoglycan (SG), loss of all four SGs, alpha-, beta-, gamma- and delta-SG proteins, and are useful for developing gene therapy of the hereditary DCM. The delta-SG is a component of dystrophin-associated glycoprotein complex that stabilizes sarcolemma. Four familial and sporadic DCM cases have been reported in human patients with the same delta-SG gene mutation. To establish the potential gene therapy of DCM, efficient and long-lasting transduction of the responsible gene is mandatory, especially for improving the functional defect. Recombinant adeno-associated virus (rAAV) vector with delta-SG gene was intramurally transfected to the TO-2 hearts at 5-weeks-old. The transfected myocardium revealed robust expression of both transcript and transgene after 10 and 20 weeks. Immunohistological analyses demonstrated re expression of not only delta-SG but also the other three SGs and normalization of the diameter of transduced cardiomyocytes without the pathogenicity. Hemodynamic studies revealed preferential amelioration of the diastolic indices. It suggests a novel strategy for the treatment of DCM and the rAAV vector is available for the treatment of several human diseases because of its safety and efficacy. PMID- 11862756 TI - [Bradykinin antagonist: current status and perspective]. AB - The kallikrein-kinin system plays an important role in many physiological and pathophysiological conditions such as homeostasis of circulation, inflammation/allergy, pain, shock, etc. Two types of kinin receptor are known, bradykinin (BK) B1 receptor and BK B2 receptor. B2 receptors are constitutively expressed and mediate most physiological actions of kinins, whereas B1 receptors are highly inducible upon inflammatory stimulation or tissue injury, suggesting that they are involved in inflammation and/or nociception. Only three peptide type B2 antagonists, NPC 567, CP-0127 and HOE-140, have been evaluated in clinical studies so far, and some beneficial effects of B2 antagonists have been shown for rhinitis, asthma, systemic inflammatory response syndrome/sepsis and brain injury. However, the results were less convincing than expected. Now several potent and orally active nonpeptide B2-receptor antagonists have been found, which are expected to overcome the weak point of the peptide type antagonists and clarify the therapeutic potential of the B2-receptor antagonist for novel indications as well as those mentioned above. As for B1 receptors, no antagonist has been tested in a clinical trial. The important role of B1 receptors is just being elucidated by use of peptide type antagonists or B1 receptor gene knockout mice. The further development of newer B1 antagonists and clinical evaluation is desired. PMID- 11862757 TI - [The arginine paradox]. AB - L-Arginine has attracted major interest because it has been identified as the natural substrate of nitric oxide synthase and is now recognized as a major player in the regulation of biological function. The arginine paradox refers to the phenomenon that exogenous L-arginine causes NO-mediated biological effects despite the fact that nitric oxide synthases (NOS) are theoretically saturated with the substrate L-arginine. There have been several explanations for this phenomenon, although none of them can explain the arginine paradox fully: (1) L arginine-induced insulin, which has vasodilatory actions. (2) Neither extracellular nor intracellular concentration determines the NOS activity but rather the L-arginine amount transported across the plasma membrane may do so. (3) Endogenous NOS inhibitors reduce the enzyme sensitivity to L-arginine. These inhibitors include, NG, NG-dimethyl-L-arginine, L-citrulline, argininosuccinic acid and agmatine. (4) Intracellular L-citrulline, an NOS product, is a potent inhibitor of NOS so that the cells may need extra L-arginine to compete with L citrulline inhibition. PMID- 11862758 TI - [Pharmacological and clinical profile of nifekalant (shinbit injection), a class III antiarrhythmic drug]. AB - Nifekalant (shinbit, MS-551) is a pure class III antiarrhythmic drug (Vaughan Williams' classification), which was approved in Japan in June 1999. This drug prolongs the action potential duration (APD) and the effective refractory period (ERP) in cardiac myocytes mainly by blocking the IKr (the rapid component of delayed rectifier K+ current). The antiarrhythmic efficacy depends on prolongation of ERP. The importance of this drug is to save patients from the life-threatening arrhythmias ventricular tachycardia (VT) and fibrillation (VF). Nifekalant was effective against reentrant arrhythmias such as VT and VF in postinfarction dogs. This drug does not have the negative inotropic effect that has been observed with other antiarrhythmic drugs. In clinical therapy, this drug was remarkably effective on patients who were unresponsive to therapy with other drugs or who were not able to receive other drugs due to decreased cardiac function. There has been no case of drug-induced worsening of the cardiac function. The significant adverse reaction is proarrhythmia such as ventricular tachycardia including TdP. Nifekalant is expected to be a useful drug for patients who could not be rescued from life-threatening arrhythmia by conventional therapies. PMID- 11862759 TI - [Pharmacological profile and clinical effect of zolpidem (Myslee tablets), a hypnotic agent]. AB - Zolpidem is a non-benzodiazepine hypnotic agent with a chemical structure of imidazopyridine. In vitro and in vivo binding studies, zolpidem exhibits selectivity to omega 1 receptors (GABAA-receptor subtypes containing alpha 1 subunits). Unlike benzodiazepines, zolpidem produces sedative effects in preference to anxiolytic, anticonvulsant and myorelaxant effects in behavioral experiments using mice. Double-blind comparative studies with reference drugs such as triazolam and zopiclone show that zolpidem is an effective and highly safe drug for the treatment of insomnia. In addition, zolpidem does not produce next-day residual effects, rebound insomnia and tolerance. This clinical profile of zolpidem may be related to its selectivity and high intrinsic activity for omega 1 receptors. PMID- 11862760 TI - [Role of protein kinase C isozymes in cellular functions and pathological conditions]. AB - Protein kinase C (PKC) is a superfamily of lipid-dependent protein Ser/Thr kinases consisting of at least 10 isozymes. The present article summarizes the papers presented at the congress symposium of the 74th Annual Meeting of the Japanese Pharmacological Society, in which six special topics regarding PKC isozyme-dependent cellular functions and pathological disorders were discussed. Using a GFP-tagged PKC expression technique, each PKC subtype was suggested to vary its targeting-site in each cell in response to each stimulus and that the targeting to the specific compartment is necessary for the specific cellular responses (NS). A cardioprotective agent, JTV519, was shown to attenuate post ischemic myocardial injury by mimicking ischemic preconditioning through specific activation of PKC delta (YK). Using an antisense technique, PKC alpha and delta/epsilon were shown to be necessary for gene expression of inducible NO synthase by interleukin-1, one of the proinflammatory cytokines, by a stimulated transactivation of NF-kappa B (TH). In canine cerebral artery, PKC delta and PKC alpha play important roles in the development and the maintenance of vasospasm induced by subarachnoid hemorrhage, respectively (SN); and stretch-induced MLC20 phosphorylation involves MLCK and PKC alpha but not PKC delta activities facilitated by inactivation of myosin phosphatase through Rho activity (KO & KN). To clarify the role of PKC isozymes in insulin resistance, the effects of insulin on glucose uptake, PKC isozyme activation and PI3K activation in rat adipocytes were shown and then platelet PKC beta activation in diabetic patients with various diabetic complications, including diabetic retinopathy, was reported (TI). These studies will promisingly open the way to a new era for the development of novel drugs controlling an isozyme-specific activity of the protein kinase C superfamily and improvement in the knowledge about the role of the protein kinase in health and disease. PMID- 11862761 TI - [Neuronal death mode switch and neurogenesis as in vivo neuroprotection]. AB - The brain has various in vivo neuroprotective mechanisms that allow it to survive for an entire lifetime. As well as neurotrophic factor-mediated inhibition of in vivo apoptotic mechanisms through various protein kinases including Akt and MAP kinase, we propose adding the neuronal death mode switch mechanism observed under the brain ischemic stress to the list of neuroprotective mechanisms. Necrosis occurs when energy or ATP levels are markedly reduced. Lowered ATP levels cause a Na(+)-K(+)-ATPase failure, leading to an osmolysis. On the other hand, sufficient ATP is required for the apoptosome activation. Under the serum-free condition, cortical neurons rapidly die in necrosis. High-glucose treatment converted the cell death mode to apoptosis through an elevation of cellular ATP levels. This treatment also rescued the cell from death due to retinal ischemic injury. These findings suggest the possibility that ischemia-induced neuronal death could be inhibited by some drugs to elevate cellular ATP levels. Neurogenesis in the adult brain is now an important topic in neuroscience. As brain injury is reported to enhance the neurogenesis, this might be also included in the ways of in vivo neuroprotection. As lysophosphatidic acid has various activities to drive neurogenesis, the neurogenesis could also be managed by other drugs to compensate for functions lost by neuronal death. PMID- 11862762 TI - [Heat shock protein 27 in osteoblasts]. AB - Physiological stresses such as heat stress, chemical stress and mechanical stress induce the expression of heat shock protein (HSP) families in cells, which affects cell function. In the present review, we describe HSP27, a small HSP in osteoblasts, especially the regulatory mechanism of the induction of HSP27 stimulated by physiological bone agents. Chemical stress by sodium arsenite (arsenite) induces HSP27 coupled to the metabolic activity of the arachidonic acid cascade, and the HSP27 induction by arsenite is negatively regulated by activation of protein kinase C (PKC). On the contrary, physiological regulators of bone such as endothelin-1, prostaglandin F2 alpha (PGF2 alpha), PGD2, and basic fibroblast growth factor (bFGF) induce HSP 27 via protein kinase C (PKC) activation. In addition, the mitogen-activated protein (MAP) kinase super-family takes part in the HSP27 induction. Thus, not only stress but also physiological agonists induce HSP 27 in osteoblasts, and PKC or MAP kinases play important roles in the induction of HSP27. PMID- 11862763 TI - [Angiotensin II type 2 (AT2) receptor signal and cardiovascular action]. AB - Due to the discovery of nonpeptic ligands, the receptors for angiotensin (Ang) II are classified into two subtypes (AT1-R and AT2-R). AT1-R mediates most of the cardiovascular actions of Ang II. AT2-R is expressed at very high levels in the developing fetus. Its expression is very low in the cardiovascular system of the adult. The expression of AT2-R can be modulated by pathological states associated with tissue remodeling or inflammation. In failing hearts or neointima formation after vascular injury, AT2-R is reexpressed in cells proliferating in interstitial regions or neointima and exerts an inhibitory effect on Ang II induced mitogen signals or synthesis of extracellular matrix proteins, resulting in attenuation of the tissue remodeling. An extreme form of cell growth inhibition ends in programmed cell death, and this process, which is initiated by the withdrawal of growth factors, is also enhanced by AT2-R. Cardiac myocyte- or vascular smooth muscle-specific mice that overexpress AT2-R display an inhibition of Ang II-induced chronotropic or pressor actions, suggesting the role of AT2-R on the activity of cardiac pacemaker cells and the maintenance of vascular resistance. AT2-R also activates the kinin/nitric oxide/cGMP system in the cardiovascular and renal systems, resulting in AT2-R-mediated cardioprotection, vasodilation and pressure natriuresis. These effects, transmitted by AT2-R, are mainly exerted by stimulation of protein tyrosine or serine/threonine phosphatases in a Gi-protein-dependent manner. The expression level of AT2-R is much higher in human hearts than in rodent hearts, and the AT2-R-mediated actions are likely enhanced, especially by clinical application of AT1-R antagonists. Thus, in this review, the regulation of AT2-R expression, its cellular localization, its pathological role in cardiovascular and kidney diseases, and pharmacotherapeutic effects of AT2-R stimulation are discussed. PMID- 11862764 TI - [Gastrointestinal side effects of non-steroidal anti-inflammatory agents and costs in the National Health System]. PMID- 11862765 TI - [Severe gastrointestinal complications potentially associated with the use of non steroidal anti-inflammatory agents: hospital treatment costs for the National Health System of our country]. AB - BACKGROUND: Non-steroidal antiinflammatory drugs (NSAIDs) produce severe gastrointestinal (G-I) complications in 1-4% of cases which need to be treated into the hospital. The aim of this study has been to assess the hospital cost secondary to treat these complications in our National Health Service (NHS). MATERIAL AND METHODS: In the first phase a cross-sectional study was performed in order to know the number of patients who were hospitalized due to a severe G-I complication during 1998 in two tertiary hospital in our country. It was reviewed their clinical charts to know whether they had taken any NSAIDs. In those positive cases all resources used during the hospitalization were collected. RESULTS: In both hospitals studied 38.1% of hospitalized patients for a severe G I complication had taken any NSAIDs during the same day or previous days. The cost/patient was of 389,831 pesetas. During 1998 in the whole NHS there were 54,623 hospitalizations owing to the same reason. Assuming that 38.1% of them had also taken any NSAIDs, 20,811 patients would have suffered a severe G-I complication potentially due to the intake of NSAIDs. Extrapolating the cost/patient obtained in both hospitals to the global number of patients hospitalized into the NHS, the cost of treating all severe G-I complications related to the consumption of NSAIDs during 1998 was of 8,112 millions pesetas. CONCLUSIONS: Bearing in mind the elevated prevalence of osteo-articular pathology in our country and the high consumption of NSAIDs for its treatment, the coming of new therapeutic options with a better safety profile would mean an important resources' saving for our NHS. PMID- 11862766 TI - [Descriptive study of ambulatory patients with alcohol-related liver disease in our setting]. AB - AIMS: We tried to show the demographic characteristic and alcohol intake habits among our outpatients. We study the influence of age, sex, habitat and socioeconomical status on alcoholic habit. DESIGN: Retrospective and institution based study. Patients. 164 patients who were followed up for alcohol liver disease in our outpatient section. RESULTS: Average age to start drinking alcohol was 18.6 (7.36) years, years of alcoholism were 35.4 (13.5) years, average daily alcohol intake was 161.2 (116.7) grams of pure alcohol. Only 16 men (8%) drank less than 60 grams a day. 5 (35.7%) women drank less than 40 grams a day. Life cumulative alcohol intake was correlated with Maddrey's score at the end of the study (r = +0.407). Average daily alcohol intake was correlated with ultrasonographic features of the liver (r = +0.283), we appreciated that Prothrombin Time was also correlated with ultrasonographic features of the liver (r = +0.301). The percentage of patients who suffer, at least one decompensation of their disease was 39%. CONCLUSIONS: Average age to start drinking is about legal age. Life-cumulative alcohol intake was related to Prothrombin Time and ultrasonographic features of the liver. PMID- 11862767 TI - [Peripheral septic arthritis in adults. Epidemiologic study in a Galician health area]. AB - BACKGROUND: Both, clinical and biologic manifestations of septic arthritis are long been know, but few studies of its epidemiological aspects are well documented. Literature concerning epidemiological aspects of septic arthritis remains exceptional in Galician Autonomic Community. OBJECTIVE: The aim of this investigation was to study the etiopathogenesis and epidemiological characteristic of septic arthritis in the adult population from hospital county of Servicio Galego de Saude (SERGAS). MATERIAL AND METHODS: We conducted a retrospective study of all peripheral septic arthritis registered between 1 January 1995 and 31 December 2000 on adult population of our health district of SERGAS (127.000 inhabitants). The location, etiology, pathogenesis and epidemiological characteristics were obtained from medical records. RESULTS: The case records of 45 patients with 46 septic arthritis (37 native joints and 9 prosthetic joints) were registered. The mean age was 57 years and 24 patients were male. During the study period, the incidence of bacterial arthritis suffered a progressive reduction to 10.2/105 inhabitants (1995) up to 3.1/10(5) (2000) with a median of 7.2/10(5). Ten patients (22%), eight male and two women, had a post-traumatic septic arthritis. Major risk factors were diabetes mellitus (5 patients), rheumatoid arthritis (5 patients) and intravenous drug abuse (3 patients). Staphylococcus aureus was the principal causative agent (28 patients, 62%) and the knee was the most commonly affected joint (41%), followed by the hip (15%) and ankle (15%). CONCLUSIONS: During the past six years, the incidence of adult peripheral septic arthritis in our SERGAS health district has been gradually reduced. Trauma was a important factor in the development of septic arthritis through direct inoculation or through spread from adjacent infectious focus. The major systemic factors predisposing to joint infection were rheumatoid arthritis and diabetes mellitus. In both, native and prosthetic joints, Staphylococcus aureus was the most common isolated pathogen. PMID- 11862769 TI - [Pituitary apoplexy: retrospective study of 9 patients with hypophyseal adenoma]. AB - Pituitary apoplexy is an acute hemorrhage or ischemia infarction of the pituitary gland, almost invariably occurring in the presence of an pituitary adenoma. Although intratumoral bleeding occur in about 9.5 to 25% of pituitary adenomas, various series suggest that clinical apoplexy may be diagnosed in about 2% to 10% of the adenomas. In a retrospective study from 1988 to 1998 of 110 patients with hypophyseal adenomas, there were 9 cases with pituitary apoplexy, yielding an incidence of 8.2%. Their mean age was 52.4 +/- 12.8 anos years, with a male to female ratio of 7:2. Symptoms observed were headache (89%), sudden visual deterioration (78%), vomiting (78%) and oculomotor nerves paresis (33%). The diagnosis of pituitary apoplexy was established by computerized tomographic scans, and hypophyseal macroadenoma with intratumoral bleeding was observed in every patient. Five patients underwent transphenoidal surgery. Improvement of visual deficit was observed in 3/4 (75%) and ocular paresis in 3/3 (100%) of affected patients. Four patients were treated conservatively with steroids. Two patients who had visual deficit recovered it completely without surgery. Two hypophyseal adenomas were resolved spontaneously after bleeding, one stayed unchanged and another presented recurrence of bleeding at six years of follow-up. Steroid and thyroid hormone replacement therapy was required in 62.5% of patients. PMID- 11862768 TI - [Seasonal changes in hospitalization and mortality resulting from chronic heart failure in Vigo]. AB - OBJECTIVES: Circannual variation in morbi-mortality for ischemic heart disease is well-known but there are few data focusing on chronic heart failure (CHF). This report analyzes seasonal variations in CHF hospitalizations and mortality in Vigo, Northwest of Spain. METHODS: Data on hospital discharge reports with a diagnosis of CHF (3-digit ICD9 code 428) were obtained from the Clinical Documentation Service at Hospital Meixoeiro (419 beds, population: 167.000 inhabitants > 14 years old). Data were divided in three 4-months periods: winter (november-february, years: 1997-98, 98-99 and 99-00), spring (mars-june, years: 97, 98 and 99) and summer (july-october, years: 97, 98 and 99). Hospitalization rates and mortality were also studied at Cardiology, Internal Medicine and Geriatrics (attending to 81% of patients with CHF). RESULTS: A total of 1.472 CHF hospitalizations were registered (52% male, 83% > 65 years, mean inhospital stay: 13.8 days). Significatives winter increases were noted in global hospitalizations -4.9% (winter) vs. 3.2@1000 (summer) and overall medical services admissions 9.3% (winter) vs. 6.2@1000 (summer). These increases also were observed at specific medical services (Cardiology: +4.9%, Internal Medicine: +6.5%, Geriatrics: +3.2%). There was not seasonal differences in letality or attributable mortality for CHF. Death in patients with a diagnosis of CHF was 4 times more likely. (OR: 3.81; 95% CI: 3.28-4.42). CONCLUSIONS: There are a striking increase in winter hospitalizations for CHF. This diagnosis is associated with an excess of inhospital mortality. Preventive and therapeutic measures taking in account this observation are warranted to reduce the burden of this growing problem. PMID- 11862770 TI - [Primary Ki-1 positive anaplastic large cell non-Hodgkin's lymphoma of the lung. A case study and review of the literature]. AB - Ki-l positive anaplastic large cell non-Hodgkin's lymphomas are a recently recognized entity, unusual and characterized by the expression of CD30 antigen. The most usual clinical feature is peripheral lymphadenopathy with mediastinal sparing and extranodal disease which occurs in approximately half of the cases, with the skin as the most common site; lung, bone marrow and central nervous system involvement are uncommon. Therefore primary pulmonary Ki-l positive anaplastic large cell non-Hodgkin's lymphomas are an uncommon clinical entity. Their classification and clinical behaviour look like high grade malignancy lymphomas, that in the most cases are presented in advanced stages disease to the diagnostic. They are presented with higher incidence in young people, where prognostic is more favourable. Clinical, morphologic and inmunophenotypic features of this lymphoma type are reported. PMID- 11862771 TI - [Reversible posterior leukoencephalopathy in a patient with non-Hodgkin's lymphoma after treatment with CHOP]. AB - Reversible posterior leukoencephalopathy syndrome is a newly characterised and increasingly recognized clinico-radiologic syndrome. Underlying conditions that reportedly trigger this syndrome include hypertensive encephalopathy, eclampsia, renal failure, and immunosuppressive drug therapy with cyclosporine, tacrolimus and interferon alpha. We describe a 51-year-old woman with non-Hodgkin's lymphoma treated with conventional CHOP chemotherapy. Eight days after this treatment she developed severe headache, bilateral visual loss and focal seizures with secondary generalization. Neurologic examination showed confusion, cortical blindness, and left hemiparesis with hyperreflexia and sensory loss. A cranial T2 weighted magnetic resonance imaging revealed increased signal intensity in the occipital and frontal lobes in both hemispheres and right parietal lobe. A diagnosis of reversible posterior leukoencephalopathy was made. She presented a favourable outcome with conservative treatment with mannitol and phenytoin. A new cranial scanning showed nearly complete resolution of the abnormalities. To the best of our knowledge, this is the first case of reversible posterior leukoencephalopathy in a patient treated with standard-dose CHOP. In this patient, we confirm the theoretical pathophysiologic mechanisms suggested explaining how these drugs can cause the syndrome. PMID- 11862772 TI - [Mycotic pseudoaneurysm caused by Mycobacterium tuberculosis]. AB - Mycobacterium tuberculosis can involve any organ of the body, including the arteries. Usually the mycobacteria infects the arterial wall spreading from a contiguous foci. We report a case of tuberculous pseudoaneurysm involving the iliac artery clinically manifested as fever of unknown origin. The diagnosis relied on abdominal CT-scanning. Tuberculous etiology was confirmed postoperatively by microbiologic and microscopic study. The antituberculous therapy was early started, but the patient died three months later as a consequence of a non-infectious abdominal aortic rupture. Clinicians should consider tuberculous etiology when the diagnosis of mycotic pseudoaneurysm is being entertained. PMID- 11862773 TI - [Mental disorders in patients infected with the human immunodeficiency virus]. AB - The present review aims to offer a concise of information about the diverse mental disorders affecting HIV-infected patients. Although most studies coincide in remarking that HIV-infected patients are frequently burden with psychological distress, the prevalence of the different mental disorders being present at the time of evaluation is widely variable. HIV infection clinical stage, prior psychiatric morbidity, and sociodemographic issues are also related to the type and risk for mental disorders. When planning therapeutic interventions, psychopharmacological or psychological, for HIV-infected patients several peculiarities should be taken into account. The accurate psychosocial evaluation and prompt therapeutic intervention, could help to reduce psychiatric-psychologic morbidity in a population of patients with multifactorial impairment in their quality of life and improve the adherence to treatment. PMID- 11862774 TI - [Danazol-induced hepatitis with functional deficit of the C1 inhibitor]. PMID- 11862777 TI - [Neuroleptic malignant syndrome]. PMID- 11862775 TI - [Reset osmostat or altered osmoreceptor]. PMID- 11862778 TI - [Portal hypertension as complication of hepatic hydatidosis]. PMID- 11862779 TI - [Hypersensitivity reaction to phenytoin]. PMID- 11862780 TI - [Factitious hypoglycemia: report of a case]. PMID- 11862781 TI - [Difference depending on age decade of the impact of hospitalization on elderly patients]. PMID- 11862782 TI - [Fat embolism syndrome: report of a case and review of the literature]. PMID- 11862783 TI - [Atypical presentation of brain malaria]. PMID- 11862784 TI - [Don't be hasty in reaching for the knife! (interview by Dr. Renate Jackle)]. PMID- 11862785 TI - [Borderline values, antihypertensive drugs in comparison, additional measures. General practice tips for therapy of hypertension]. PMID- 11862786 TI - [Quincke edema emergency. Don't hesitate with the adrenaline]. PMID- 11862787 TI - [Risk of thrombosis in long distance flights. What prevents "jet leg"?]. PMID- 11862788 TI - [Poorly tolerated or genuinely allergic? How to properly assess food allergies]. AB - Massive immune reactions to certain foods may be traced back to immunological, but also to non-immunological mechanisms. The diagnostic work-up must therefore aim to distinguish an allergy from other inflammatory, neoplastic or immunoregulative disorders. The most common form of gastrointestinal-mediated allergy form are the IgE-mediated reactions of the immediate type (Type 1 allergy). The routine diagnostic investigation includes history-taking, a daily diary of foods eaten, skin tests and antibody determination. The diagnosis of an allergy is often complicated by the multitude of possible manifestation sites, a possibly localized effect, and variations in the depth of allergic processes. To confirm the diagnosis, an oral provocative test should always be performed. PMID- 11862789 TI - [Fish, olive oil, fruit, vegetables and fiber. Therapy of hypertension a la carte]. AB - A healthy diet has a positive impact both on elevated blood pressure and its associated cardiovascular risk. In particular the association between obesity and hypertension is beyond doubt. Accordingly, a calorie-controlled or low-calorie diet resulting in a reduction in weight, lowers raised blood pressure. Blood pressure is also determined by the composition of the diet. A substantial pressure-lowering effect has been shown to be associated with the regular ingestion of fish and olive oil. Evidence also points to a positive effect of a diet high in fruit and vegetables and low in fat. Also recommended for hypertensives is a low-salt and potassium-rich diet, and sparing use of alcohol. The impact of a healthy diet on the cardiovascular risk is particularly favorable when the patient also takes regular physical exercise. PMID- 11862791 TI - [Nutrition in chronic inflammatory bowl diseases. What your patient tolerates is permitted]. AB - Modifying the diet can have a favorable impact on the course of chronic inflammatory bowel disease. In contrast, nutrition plays no major role in the development of the disease or in provoking an acute attack. During an acute attack of Crohn's disease, the application of oral (drinks) or tube feeding (enteral nutrition) can result in a substantial clinical improvement. The remission rate of this side effect-free therapy is, however, lower than that seen with steroids. The data available for diet therapy in ulcerative colitis are less unequivocal, but the effects are probably slight at best. During the remission phase, the patient should eat a balanced, vitamin-rich and varied normal diet that excludes all poorly tolerated foods: lactose intolerance in particular appears to be increased in patients with Crohn's disease. Known vitamin or mineral deficiencies should be corrected by appropriate dietary measures or supplementation. There is no such thing as a "Crohn's diet" or "colitis diet". The patient can be allowed to eat anything that is tolerated. PMID- 11862790 TI - [Omega-3-fatty acids, Mediterranean cooking, low-fat diet. What really prevents myocardial infarct?]. AB - The strength of evidence in support of dietary measures in the secondary prevention of coronary artery disease is variable (Table 1). The most impressive data available concern the impact of omega-3 fatty acids found in sea-fish at an intake of about 1.0 g/day, but the data underscoring the value of a Mediterranean diet are also convincing. No comparable data are available for low-fat and fat modified diets. For diabetics, an HbA1c of < 7% should be achieved. There is indirect evidence that a body mass index of between 18.5 and 23.9 kg is optimal, and can be achieved by physical exercise and weight control. Weight reduction also lowers blood pressure, a goal that can equally be reached by a low-salt diet rich in fruit and vegetables, and the practice of endurance sports. The results of ongoing studies on the use of vitamin supplements are as yet unavailable. PMID- 11862792 TI - [Cancer-inducing alcohol. "Reasonable" drinking also causes damage]. AB - More than 75% of all types of cancer are due to environmental factors. These latter include nutrition. A diet rich in fruit and vegetables, carotenoids, and vitamin C has a protective effect against cancer. A high-calorie, high-fat diet, and the consumption of alcohol have the opposite effect. New data appear to show that the daily consumption of up to 20 g alcohol in women, and 40 g in men, previously considered innocuous, must be considered a risk for certain types of cancer. A particularly close association with alcohol is found in the case of cancer of the breast, and tumours of the mouth, pharynx and esophagus. PMID- 11862793 TI - [Oral therapy in type 2 diabetes. Critical evaluation of glitazones]. PMID- 11862795 TI - [Immunosuppression by individual need. Protection for the donor kidney]. PMID- 11862794 TI - [New triptanes in control of migraine attacks. More rapid onset of action--more efficient reduction of pain]. PMID- 11862796 TI - [Preventing rejection, minimizing toxicity: optimized therapy with cyclosporin]. PMID- 11862797 TI - [Lumbar intervertebral disk displacement. Conservative management is usually better]. PMID- 11862798 TI - [Evaluating lung disease with heart catheter? Proper management in COPD]. PMID- 11862799 TI - [Blood pressure control must be effective until the next tablet. Infarct is most frequent in the morning]. PMID- 11862802 TI - [Moxonidine for hypertension. "A good base for combination therapy"]. PMID- 11862800 TI - [ESC guidelines for therapy of heart failure. AT1 blockers are now also included]. PMID- 11862801 TI - ["Critical dose" drugs. Aut-idem could cause problems here]. PMID- 11862803 TI - [Coronary heart disease patient with hypertension. Dangerously increased blood pressure despite gigantic therapy arsenal]. PMID- 11862804 TI - [28. Acute therapy and prevention of migraine]. PMID- 11862805 TI - 2002 overview: the role of activism. PMID- 11862806 TI - Heart disease prevention. PMID- 11862807 TI - Drug interactions need more attention. PMID- 11862808 TI - More uses for tenofovir? PMID- 11862809 TI - New kinds of treatment. PMID- 11862810 TI - Legal/medical issues. PMID- 11862811 TI - Drug pricing. PMID- 11862813 TI - Improving fundraising. PMID- 11862812 TI - Improving activism. PMID- 11862814 TI - Funding medical research and drug development. PMID- 11862816 TI - Keeping OR suite going during construction is a tall order. PMID- 11862815 TI - Diet and inflammation? (Personal note). PMID- 11862817 TI - Surgery for the morbidly obese requires a special commitment. PMID- 11862819 TI - Techniques for obesity surgery. PMID- 11862818 TI - Increase in bariatric surgery brings a surge in legal cases. PMID- 11862820 TI - Every project needs infection risk plan. PMID- 11862821 TI - Lessons learned through OR renovation. PMID- 11862822 TI - Ten JCAHO standards that trip up ASCs. PMID- 11862823 TI - Practice guidelines: what JCAHO expects. PMID- 11862825 TI - Basic mammalian bone anatomy and healing. AB - The goal of any method of fracture repair should be the early return to function of the patient with minimum postoperative morbidity. This is accomplished most optimally by having a basic understanding of the biology of bone healing and by being familiar with the musculoskeletal system of the species before attempting fracture repair. Applying the fundamental principles of mammalian bone anatomy and physiology to the bird ensures the best prognosis possible and minimizes postoperative complications in the avian patient. PMID- 11862824 TI - Orthopedic radiography in exotic animal practice. AB - Radiographic examination of exotic animal species provides significant diagnostic information to the practitioner so long as attention is paid to positioning and radiographic detail and if variations in normal radiographic anatomy are contemplated. Fracture of pneumatic bones (humerus or femur) in avian patients often results in subcutaneous emphysema, much like an open fracture. If fracture alignment is anatomical, there is generally little callous formation visible in avian and reptile patients; however, if malunion occurs, then certainly callous can be visualized. Aggressive or excessive periosteal proliferation in exotic species often predicts osteomyelitis. Practitioners can increase their diagnostic capabilities and thereby decrease any preconceived anxiety by applying the same general interpretive concepts used in the radiography of more routine small animal patients and by evaluating the resultant radiographs in a systematic manner. With the exception of certain anatomical differences and alterations in response to injury, exotic animal species suffer the same types of orthopedic disease do dogs and cats. PMID- 11862826 TI - Ferret orthopedics. AB - Orthopedic conditions requiring surgical intervention and correction frequently occur in the ferret patient. Elbow luxations and long bone fractures are among the most common syndromes affecting the ferret patient, which often presents in an emergency setting. Orthopedic conditions in the ferret, as in other mammalian species, are often the result of trauma. Surgical treatment including reduction and stabilization of the affected bony structure, combined with supervision and the recommended postoperative care, can provide a good prognosis to return to normal function for the ferret. PMID- 11862827 TI - Rabbit orthopedic surgery. AB - Orthopedic surgery in rabbits poses several unique parameters for the veterinary surgeon. It is imperative for the veterinarian to be knowledgeable about the anatomic features of the surgical repair site and to become familiar with a rabbit's pain and discomfort often associated with orthopedic injuries. Handling the perioperative and postoperative pain and potential GI disturbances are crucial for a successful outcome of the surgical case. This article is designed to help the veterinary surgeon prepare for the orthopedic surgical procedure and the peripheral physiologic needs of the rabbit from presentation through recovery. PMID- 11862828 TI - Small exotic mammal orthopedics. AB - The basic principles of orthopedic repair are the same for small exotic mammals as for the canine or feline patient. This article assists the clinician in determining the most appropriate method of fixation and maximizes the probability of a favorable outcome. PMID- 11862829 TI - Postoperative management of the exotic animal patient. AB - Careful postoperative management is crucial for the success of any orthopedic surgical repair. The special needs of the exotic animal must be met during the immediate postoperative period and during hospitalization. Many exotic animals require a quiet, stress-free environment and careful attention to housing parameters, such as perching, temperature, and visual security. To provide adequate pain relief in these patients, analgesia should be given before, during, and after surgery. The drugs most often used for pain relief are NSAIDs and opioid analgesics. After surgery, monitor the healing process carefully with regular examinations and radiographs while remaining vigilant for potential problems such as osteomyelitis or nonunion. Physical therapy prevents the development of fracture disease, which includes joint or ligament contracture and bone or muscle mass loss. Because physical therapy affords the patient full use of the affected limb, it is considered a helpful practice in all patients. Physical therapy, however, is critical for free-ranging exotic animals that will be released back into the wild, such as birds of prey. PMID- 11862830 TI - Basic avian bone growth and healing. AB - There have been few studies on the process of fracture repair in avian species. Most of the information shows similarities between avian and mammalian bone growth and fracture repair, but there are differences. The main finding confirms that fractures must be reduced properly, stabilized, and immobilized with an adequate blood supply to the bone fragments for optimal healing. The return to function of extremities, particularly the legs and wings, is an important consideration when internal fixation methods are used. Causing little or no collateral damage to soft tissue and joint when implanting internal hardware is ideal and reduces the likelihood of impaired function. Whether internal or external fixation methods are used for fracture reduction, the knowledge of avian bone growth and fracture repair is essential for veterinarian understanding and when discussing the healing process with clients. PMID- 11862831 TI - Clinical considerations of the thoracic limb. AB - The avian skeleton is significantly different from the mammalian skeleton, and these differences are important when providing fracture repair and postoperative care for the avian patient. Successful repair depends on an understanding of fracture management and healing and of the anatomical, physiologic, and behavioral concerns of avian patients. Having knowledge of fracture repair without an understanding of the unique needs of avian patients' results in a disappointing treatment response. PMID- 11862832 TI - Orthopedic conditions that affect the avian pelvic limb. AB - Birds are often presented to the veterinarian with injury or disease that affects their ability to use one or both legs. Although in wild birds traumatic injuries usually involve the wing, captive birds especially falconer's birds, are usually presented with injuries to their legs. This article focuses on conditions affecting practice: birds of prey, and injured wild birds. The birds are presented with lameness or loss of use of the leg. The conditions described can be initiated by injury or by disease affecting an area other than the leg, such as the kidney or spinal cord, or they can be toxic or metabolic. PMID- 11862833 TI - Orthopedic conditions of the avian head. AB - Orthopedic problems of the avian head generally fall into two main categories: congenital and traumatic. Congenital lesions of the beak are not uncommon in psittacine birds but are extremely rare in raptors. Trauma accounts for most of the remaining orthopedic problems seen in the area of the body. This article discusses the most common conditions and injuries causing orthopedic problems of the beak, eye, and skull of avian patients. PMID- 11862834 TI - Diagnosis and management of reptile orthopedic injuries. AB - As veterinarians expand their understanding of the specific husbandry requirements for captive reptiles, nutritionally associated orthopedic injuries should decrease. Orthopedic injuries in wild reptiles, however, will continue to increase as new infrastructure encroaches on the habitats of these animals. Research should be pursued that focuses on improving our understanding of pain management in reptiles, on developing techniques to expedite bone healing, and on creating new orthopedic techniques that provide rigid stabilization without the use of temperature-sensitive materials. PMID- 11862835 TI - Making up your mind about mammograms. PMID- 11862836 TI - I'm a 65-year-old woman in good health. What vitamins should I be taking? PMID- 11862837 TI - I may be hospitalized soon, and my primary care doctor said that a "hospitalist" may be in charge of my care while I'm there. What do hospitalists do? PMID- 11862838 TI - Building a better food pyramid. PMID- 11862839 TI - Slipped disk discussions. PMID- 11862840 TI - Better bereavement: coping with the death of a spouse. PMID- 11862841 TI - Can bypass patients bypass big hospitals? PMID- 11862842 TI - Cancer risk declines with regular vigorous exercise. PMID- 11862843 TI - A leisurely approach to averting Alzheimer's. PMID- 11862844 TI - Choosing the right remedy for cold symptoms. PMID- 11862845 TI - Triglycerides boost risk of stroke. PMID- 11862846 TI - Many women have undiagnosed bone loss. PMID- 11862848 TI - HRT patches: ok for women with fibroids? PMID- 11862847 TI - High red-meat intake, obesity linked to cancer. PMID- 11862849 TI - Materials for restoration of primary teeth: I. Conventional materials and early glass ionomers. AB - This paper demonstrates how the treatment of primary dentition may present the clinician with increased difficulties compared with the preparation and placement of restorations in adult dentition. Established dental materials (dental amalgam and conventional glass ionomer cements) and less well established alternative materials (copper cements) are reviewed. The use of amalgam to restore primary dentition is the subject of concern amongst the dental profession in terms of lack of adhesion and potential toxicity concerns, while the low tensile strength of traditional glass ionomer cements make them less suitable for the restoration of primary dentition. PMID- 11862850 TI - Biological guides to the positioning of the artificial teeth in complete dentures. AB - Setting teeth for complete dentures is traditionally done away from the clinic in the dental laboratory. This has unwittingly given the impression that arranging tooth position is a mechanical process in which the clinician has little say. Many technicians are given few instructions, but a detailed prescription is crucial to the success of the denture. This article describes those considerations the dentist should address in communicating with the laboratory technician. A 'denture space' impression technique is described to assist the dentist in the correct prescription for posterior teeth placement. PMID- 11862851 TI - The two by four appliance: a versatile appliance. AB - The 2 x 4 appliance comprises bonds on the maxillary incisors, bands on the first permanent maxillary molars and a continuous archwire. The appliance is used in the early mixed dentition for treatment of both anterior crossbites and alignment of ectopic incisors. Four cases using this appliance are presented. This appliance offers many advantages over alternative techniques as it provides complete control of anterior tooth position, is extremely well tolerated, requires no adjustment by the patient and allows accurate and rapid positioning of the teeth. PMID- 11862852 TI - Predictable resin-bonded bridges in general dental practice. AB - This paper discusses the use of resin-bonded bridges as predictable restorations in general dental practice. The importance and relevance of case selection is stressed. Efforts have been made to discuss ideal design features, creation of interocclusal space and the need to carry out any tooth preparations. In addition, the clinical stages involved are outlined and a brief comparison with conventional bridges and implant-retained prostheses made. PMID- 11862853 TI - Osteomyelitis complicating osteopetrosis--a case report. AB - Osteomyelitis of the jaws is uncommon but can be a complication of extraction or exposure of teeth in patients who are immunosuppressed or have undergone radiotherapy to the head and neck region. We report a case of osteomyelitis in a patient who has osteopetrosis. The initial clinical presentation was that of an infected retained root, secondary to denture trauma to the supporting tissues. Radiological interpretation was difficult owing to the increased bone density and surgical exploration revealed only a dense region of necrotic bone. PMID- 11862854 TI - Use of a matrix to form directly applied resin composite to restore worn anterior teeth. AB - The technique of using directly applied resin composite is increasingly being chosen to restore worn anterior teeth. This paper describes the use of a vacuum formed matrix ('suck-down') to shape the directly applied resin composite, avoiding demanding freehand build-ups. PMID- 11862855 TI - Multiple sclerosis presenting to the dental practitioner: a report of two cases. AB - In this paper, two patients are described who presented with trigeminal neuralgia which turned out to be an early symptom of multiple sclerosis. General dental practitioners need to be aware of the possible causes for atypical facial pain, especially in patients under 40 years. PMID- 11862856 TI - Do we need practitioner training schemes in orthodontics? AB - Orthodontic treatment appears to be in ever-increasing demand, but the number of specialists is insufficient to meet the current need. This article considers the role of the extended practitioner training scheme in the present climate of uncertainty surrounding orthodontics in the general dental services in the UK, particularly for the general dental practitioner. PMID- 11862857 TI - Ethics and dentistry: 2. Ethics and risk management. AB - The previous paper explored the meaning of ethics, especially its relationship to dentistry. Here, we examine a practical application for solving ethical problems. Together, the two articles should provide dentists with a core of relevant knowledge about ethics and a ready guide to the daily relevance of ethics. PMID- 11862858 TI - The greatest gift: volunteering. PMID- 11862859 TI - Seeking the "uneventful" with parenteral and enteral: a clinical update and synopsis of the rules and regulations governing parental and enteral conscious sedation in the State of Texas. PMID- 11862861 TI - Oral and maxillofacial pathology case of the month. Proliferative periostitis. PMID- 11862862 TI - Testing your diagnostic skills (#56). Case No. 1. PMID- 11862863 TI - Testing your diagnostic skills (#56). Case No. 2. PMID- 11862864 TI - Get informed consent before beginning treatment. PMID- 11862866 TI - Re: Fixed lingual retention and relapse. PMID- 11862865 TI - Varying treatments of Class II malocclusions: two case reports. AB - Two female patients, JH aged 13 years 5 months and CN, aged 13 years 10 months, presented at the Sydney University Orthodontic Clinic, Australia. Patient JH complained that her lower teeth were "too far back behind her top teeth". She was treated using the Speed Appliance. Patient CN complained that her teeth were "crooked". The main point of interest of her case is the mechanics used following the extraction of her four first molar teeth. Both patients were keen to improve their appearance. PMID- 11862867 TI - Continued interest in the stability of orthodontic cases. PMID- 11862868 TI - A new bracket system: Control 21. Part II: Tooth movement. AB - The Control 21 bracket system has several attributes that overcome many of the friction, binding, notching and management problems encountered with other bracket systems. The combination of a freely-moving slide in a base connected by an elastomer provides the basis for the new concept of a "bracket engine": a source of low magnitude forces able to release friction and binding forces. The "bracket engine" is continually energized by functional forces that open or close the bracket slot. The bracket's inherent elasticity reduces the severity of high forces that can damage periodontal tissue. This new concept of an active, dynamic bracket system will require a paradigm shift in thinking away from the current concept of the bracket as a passive participant in tooth movement. The mechanisms that create these attributes are explained and some potential developments are noted. PMID- 11862869 TI - Overbite correction and sagittal changes: late mixed-dentition treatment effects. AB - This study involved the assessment of pre- and post-treatment lateral cephalograms from 182 late mixed-dentition patients with Class I and Class II malocclusions, with or without crowding, selected before treatment commenced. It was designed specifically to determine whether antero-posterior mandibular dento alveolar and skeletal changes occurring with growth and one particular late mixed dentition treatment approach are influenced in any way by the required amount of incisal overbite correction to be achieved with treatment. Significantly greater forward movements were found at both point B and pogonion in non-headgear patients in whom very deep overbites had been reduced, when compared with patients with less required incisal overbite reduction. This finding would seem to have provided further quantitative evidence to support the concept of unlocking of dento-alveolar structures with early reduction of very deep incisal overbites in appropriate patients. The suggestion was therefore made that the bite-opening in such deep-bite patients should be undertaken at least as early as this late mixed-dentition stage in order to provide an improved environment for the mandibular dento-alveolus as the mandible itself moves forward with normal growth. PMID- 11862870 TI - Induced mandibular condylar growth in a sheep model after functional appliance treatment. AB - This paper reports on changes in mandibular condylar growth in sheep fitted with experimental functional appliances. The paper also addresses the similarities of mandibular condylar growth in sheep and humans. Experimental functional appliances were developed, constructed and applied to four, four-month old, male sheep. Another four sheep served as matched controls. All animals received fluorochrome bone labels during the study and their weight gain also was recorded. Fifteen weeks later, all animals were sacrificed and each temporo mandibular joint and left metacarpus were removed. Undecalcified mid-sagittal sections of the joint and metacarpus were prepared for fluorescence microscopy. The mandibular condylar growth vector in sheep was found to be in a postero superior direction. Condylar growth in the control sheep during the experimental period varied from 8.8 to 11.9 mm, with the mean being 10.6 mm, which is quantitatively similar to two years of condylar growth in human adolescents. In the experimental sheep, the condylar growth varied from 8.5 to 13.3 mm, with the mean being 11.4 mm. When metacarpal growth and weight gain were taken into consideration using multivariant analysis, the coefficients for growth in the postero-superior and posterior direction were found to be high, with adjusted r2 as 0.84 and 0.82 respectively. The induced condylar growth was estimated to be largest in the posterior direction (2.30 mm), which is also similar to previous reports in humans. These findings suggest the appropriateness of using sheep as a model for quantitative histological analysis when investigating the mechanisms of functional appliance treatment. PMID- 11862871 TI - Finishing occlusion in Class II or Class III molar relation: therapeutic Class II and III. AB - The most frequent extraction regime consists of the removal of upper and lower premolars. Depending on anchorage requirements, camouflage treatment options, surgical intervention, or the absence of teeth in only one arch, it may become necessary to finalize the occlusion with a one-dental-unit discrepancy between the upper and lower dental arches. Guidelines are presented for finishing occlusions in Class II or Class III molar relation. PMID- 11862872 TI - Profile of orthodontic services provided in private practice by general practitioners and specialist orthodontists for an insured population. AB - In Australia, the proportion, volume and type of orthodontic care provided by general practitioners and orthodontists are largely unknown. The overall objective of this study was to create a profile of orthodontic services provided by general practitioners and orthodontists for a cohort of insured patients using private practices in New South Wales, Australia, from 1st January 1992 to 31st December 1995. Data were derived from claims records submitted by members of a health insurance fund for rebates for fees paid to private practitioners for orthodontic services they received during the study period. Distribution of the volume and type of service provided by general practitioners and orthodontists was compared using the Chi-squared test. Statistical significance was taken at the 5 per cent level. Orthodontic services were provided predominantly by orthodontists (80%). Fixed orthodontic treatment was provided almost exclusively (91%) by orthodontists. The majority of removable appliance services was provided by general practitioners. Orthodontists provided more orthodontic services in the capital city and other metropolitan areas, whereas general practitioners provided more orthodontic services in rural areas. Orthodontists provided more services to members in the highest socio-economic group, whereas general practitioners provided more services to members in the lowest socio-economic group. PMID- 11862873 TI - Factors associated with administration route when using midazolam for pediatric conscious sedation. AB - Midazolam conscious sedation records of pediatric dental patients, one to six years of age, were reviewed retrospectively to: 1) examine the factors associated with the use of oral and nasal routes of administration and their effect on displayed behavior during dental treatment and 2) determine whether a child's compliance with oral administration is predictive of the intraoperative behavior displayed during dental treatment. Two hundred and fifty-seven conscious sedation records for 222 pediatric dental patients sedated with orally or nasally administered midazolam for dental treatment at the University of Florida were reviewed. Data collected included the patient's age, gender, route of administration, dose, compliance with oral administration, appointment type (planned vs. emergency), previous sedation experience with midazolam, operator vs. parent administration of the medication, use of papoose board and nitrous oxide/oxygen inhalation, types of procedures performed (restorations only, extractions only, or both), length of treatment rendered, and preoperative and intraoperative behavioral assessments of the child. The collected data were analyzed with Statview software using ANOVA and Chi-square analyses. There was a statistically significant difference (p < 0.001) between oral and nasal administration for the parameters of age, procedure length, appointment type, procedures performed, previous sedation experience and use of the papoose board and nitrous oxide/oxygen inhalation. While there was no statistically significant influence of chronological age on the preoperative Frankl behavior ratings, there was a statistically significant mean age difference with respect to the administrator of the medication (parent vs. operator), papoose board use, N2O/O2 use and previous sedation experience. Forty-five percent of the subjects were willing to accept oral administration of the medication, however, there was no statistically significant difference (p = 0.114) between the child's compliance to accept the medication and the intraoperative Frankl behavioral ratings displayed during dental treatment. In this review of midazolam conscious sedation records of pediatric dental patients. 1) route of administration was significantly influenced by several patient and procedural variables, resulting in different behavioral outcomes and 2) compliance with oral administration was not predictive of behavior displayed during treatment. PMID- 11862874 TI - Bilateral talon cusps in primary teeth: clinical significance and treatment. AB - Bilateral primary talon cusps are uncommon. This case report documents two new cases of bilateral talon cusps in the primary dentition. The clinical features, radiographic characteristics, and differential diagnosis of this developmental anomaly are described. The terminology, epidemiological data, potential complications, and various modes of treatment are reviewed and discussed. A new classification system was proposed. The dental anomalies of these two cases were not associated with any other somatic or dental abnormality. The general prognosis of this abnormality is satisfactorily good. However, the dentist needs to have early recognition of this anomaly in order to provide timely treatment in order to prevent potential complications. PMID- 11862875 TI - A study on the eruption timing of primary teeth in Korean children. AB - The objectives of this study were to clarify the eruption time and sequence of primary teeth in Korean children. A random sample of 1070 children from ages of four to thirty-six months was examined: 567 were males and 503 were females. The median values to measure the eruption time of the primary teeth were used. The results show that the primary teeth of boys erupt earlier than those of girls. As to the eruption sequence of primary teeth, the mandibular central incisor and second molar erupted earlier than their maxillary counterparts. For the lateral incisor, the canine and the first molar, the maxillary teeth erupted before the mandibular ones. PMID- 11862876 TI - Does clinical experience affect occlusal caries diagnosis and sealant recommendation? An in vitro study. AB - The purpose of this in vitro study was to evaluate the effect of the length of clinical experience on clinician's diagnosing caries and recommending sealants on the occlusal surfaces of permanent/caries free third molars. The third molars involved in this study had been extracted prior to eruption. There were three study groups: dental students, clinicians with five to ten years experience, and clinicians with fifteen to twenty years of practice. Twenty-four percent of the entire sample diagnosed caries. Clinicians with five to ten years of practice diagnosed occlusal caries significantly more than clinicians with fifteen to twenty years of practice, but not significantly differently than senior dental students. Clinicians recommended sealants significantly more than dental students. The most frequent reason for sealant indication was prevention. The results indicate that clinical experience alone does not impart better caries diagnosis and sealant recommendation criteria. PMID- 11862877 TI - Dental screening of pediatric cardiac surgical patients. AB - Dental screenings were completed on 209 pediatric patients six months and older that were scheduled for cardiac surgery from two different geographical locations. The screening involved an educational session on bacterial endocarditis and preventive dentistry followed by a dental examination. We found that sixty-two (30 percent) patients had seen a dentist regularly, ninety-three (44 percent) practiced daily oral hygiene and thirty-seven (18 percent) knew about bacterial endocarditis. Local dentists performed invasive procedures on seventy-one (34 percent) patients and thirty-four (48 percent) received antibiotic prophylaxis. Only twenty-three (68 percent) of the thirty-four patients received the then current American Heart Association antibiotic regimen. Dental disease was diagnosed in 175 (84 percent) of the 209 patients: 164 (78 percent) gingivitis; sixty (29 percent) caries; six (7 percent) dental abscess; three (1 percent) periodontal abscess; five (2 percent) periocornitis. Cardiac surgery was postponed in twenty-four (12 percent) patients. We conclude that parents lacked knowledge about bacterial endocarditis even after being informed during their routine cardiology visit, there appears to be a deficiency of knowledge among dentists regarding the indications and antibiotic regimen required to prevent bacterial endocarditis, and all cardiac surgical patients should have a dental exam and preventive dental program implemented prior to six months of age and dental screening prior to their cardiac surgery. PMID- 11862878 TI - Prevalence of cheese molars in eleven-year-old Dutch children. AB - In the Netherlands, first permanent molars with idiopathic enamel disturbances (IED) are called cheese molars. Though concern is expressed about their prevalence, adequate figures on the subject are missing. The porous enamel of cheese molars can be very sensitive to cold air and can decay rapidly. The aim of the present study was to investigate the prevalence in eleven-year-old Dutch children of cheese molars (IED). During an epidemiological study performed in four cities in the Netherlands, the first permanent molars and central incisors of eleven-year-old children were examined for hypoplasia, opacities, posteruptive enamel loss, premature extraction, and atypical restorations. The observation of a hypoplasia excluded the possibility of cheese molar. A total of 497 children were examined. Six percent (n = 128) of the molars (n = 1988) showed signs of IED (cheese molars), 10 percent of the children had cheese molars of which 8 percent two or more. Incisors (4 percent) with opacities were found in 3 percent in combination with two or more cheese molars. Among the four cities, no significant differences in occurrence were found. The results of this study showed that in 10 percent of the Dutch children eleven years of age, cheese molars (IED) were found. The cause for the phenomenon called cheese molars appears to be child centered. Further studies on prevalence, causes and prevention are mandatory. PMID- 11862879 TI - Dental disease indices and caries related oral microflora in children with phenylketonuria. AB - The purpose of this study was to investigate the dental caries, plaque and gingivitis indices and caries related oral flora in children with classic phenylketonuria. Forty-one children at The Great Ormond Street Hospital for Children and controls were included in the study. The main findings were: No significant difference in the decayed, missing and filled surfaces of the primary and permanent teeth between the phenylketonuria children and the controls. A significantly greater mean plaque score for the control children compared with the phenylketonuria children (p < 0.01) for the permanent teeth only. A significantly greater number of white opacities in the permanent teeth of the phenylketonuria group compared with the control group (p < 0.02). No significant differences in the caries related microflora. PMID- 11862880 TI - An aggressive dentigerous cyst in a seven-year-old child. AB - A review of literature indicates that very few of these cysts are found in children less than ten years of age, in spite of the fact that the cysts are associated with the eruption and/or formation of the secondary dentition. Although dentigerous cysts are rare in the first decade of life, a thorough review of a child's initial radiographs is of obvious importance as indicated by this case report. Of prime importance in this matter is the age at which the initial panoramic radiographic examination is performed on a child. According to the guidelines recommended by the American Academy of Pediatric Dentistry, the first panoramic radiographic examination should be performed following the eruption of the first permanent tooth. In this case, the appropriate radiographic survey was not conducted. A proper survey should include: Proper exposure of the film. Proper development of the film. Proper interpretation of the film. This child had a quadrant of dentistry performed three months prior to the discovery of the lesion. The previous dental provider noted that a panoramic radiograph had been exposed. It is unclear whether all of the aforementioned steps were followed or completed in order for the provider to obtain the information needed to render a diagnosis, and to properly refer this patient for a more in-depth examination of her mandibular swelling. The mere fact that this child's right mandibular first molar was fully erupted and the left one had not erupted at all, would be reason enough to take a panoramic radiograph to investigate the cause of the clinical absence of the contralateral first molar. This article has presented a case in which a proper panoramic radiographic examination performed at the time of eruption of the first permanent tooth may have led to an earlier diagnosis of the cyst, and thus might have prevented the loss of multiple permanent teeth. PMID- 11862881 TI - Candidate genes for nonsyndromic cleft lip and palate. AB - The confirmation of supposed genes causing susceptibility to nonsyndromic cleft lip with or without a cleft palate (CL/P) and cleft palate alone (CP) would help to understand the molecular development of the lip, the palate and the jaw, and to predict more accurately the risk of recurrence of such defects. The purpose of this article is to present a brief review of the current state of knowledge regarding the search for genes responsible for the occurrence of CL/P and CP. After ten years of research, approximately twenty candidate genes have already been suggested for CL/P and CP. Some genes or chromosomal regions seem to be frequently associated with these defects and a specific nomenclature (orofacial cleft genes--OFC) has been suggested. Everything indicates that it will still take many years of research before the genes responsible for CL/P and CP are confirmed. These efforts are justified, however, because of the possibility of being able to predict more accurately the risk of such defects occurring. PMID- 11862882 TI - Cultural diversity: caring for minority children with mental retardation and other disabilities. AB - Providing services for minority group children with mental retardation and other disabilities can challenge the abilities of any dental practitioner. The added challenge, however, is for the practitioner to recognize and understand the different cultures which can and do affect the relationships between the practitioner, the patient, the family, and the care that is provided. An introduction is provided to these complex and diverse minority group family settings which increasingly are becoming a component of our communities. PMID- 11862883 TI - Computerization is not the future--it is the present. PMID- 11862884 TI - Wear evaluation of MZ100 compared to ceramic CAD/CAM materials. AB - The material and antagonist wear of two CAD/CAM ceramic materials, the laboratory processed IPS Empress ceramic, and the composite mill block material MZ100 were tested in an artificial wear simulator with human enamel as the antagonistic material. The material samples underwent 50,000 test cycles (1 cycle per second, 50 N) in distilled water. The wear of the material samples and of the opposing enamel was documented after 30,000 and 50,000 cycles with replicas, digitized, and evaluated with a 3D evaluation system. The loss of volume and height caused by wear was calculated. No significant difference between the ceramic materials could be found either in the amount of material or in the amount of antagonist wear. The material wear of MZ100 differed significantly from Vita Mark II only in terms of volume loss. Regarding height loss, MZ100 exhibited a significantly higher wear than all ceramic materials. The wear of the enamel antagonists does not differ significantly between the ceramic materials. With MZ100, however, a significantly smaller amount of enamel wear was found when compared with Empress and Vita Mark II. Despite the highest material wear, MZ100 had the lowest material wear rate, the lowest enamel wear rate, and the lowest total wear rate. The laboratory-processed IPS Empress material had a clearly higher material wear rate than the CAD/CAM materials. PMID- 11862885 TI - Cerec: correlation, an accurate and practical method for occlusal reconstruction. AB - The correlation technique explained here shows one of the possibilities for occlusal reconstruction offered by the Cerec approach. The various stages of this technique are described and illustrated. The most current applications are reviewed. PMID- 11862886 TI - LAVA--the system for all-ceramic ZrO2 crown and bridge frameworks. AB - All-ceramic restorations in the posterior region are an increasingly important area of dental care. However, no real suitable ceramics or economic processing procedures have been available so far. With the new LAVA system, it will be possible to satisfy these demands in the future. The system is based on the machining of presintered zirconia, which, due to its outstanding mechanical properties, its biocompatibility, and its excellent esthetics in combination with a specially designed veneer ceramic, is the ideal candidate for these applications. In combination with a corresponding CAD/CAM unit, the use of an easy-to-machine presintered ceramic material (which is sintered to full density after shaping, thus eliminating the need for extensive use of diamond tools) allows for the first time reliable, fully-automated and thus fast manufacturing of such restorations. PMID- 11862887 TI - "Function" with registration: simple, fast, and safe using a new registration material. AB - Although the Function construction method with Cerec 2 always remained a method for enthusiastic experts, Cerec 3 enables the practiced "normal user" to use this method expediently. With the new R-Sl-LINE Metal-Bite registration material, an important step towards accelerating the process has been taken by eliminating a powder sequence and by its fast curing. Due to the special definition properties of the material, including the antagonist situation in the design of the occlusion has become considerably more reliable. PMID- 11862888 TI - Protecting our children from tobacco and alcohol. PMID- 11862889 TI - Adolescent substance use and abuse: current issues. AB - Substance use and abuse among American adolescents account for significant morbidity and mortality in this age group. A good history is the mainstay in the evaluation, along with the identification of clinical clues of drug use or abuse. Issues of consent and confidentiality should be considered in the assessment, particularly when deciding whether to use drug testing in diagnosis and management. Establishing the patient's stage of substance use can help define the option of either management by the primary care physician or referral to a professional who is experienced in treating chemical dependency. Important research-based principles for prevention programs for children and adolescents include the need for programs to target all forms of drug abuse and to be age specific, developmentally appropriate, culturally sensitive, and cost-effective. PMID- 11862891 TI - Performance-enhancing substances in adolescent athletes. AB - Use of performance-enhancing substances is widespread among adolescents. Anabolic steroids, creatine, and androstenedione are currently among the most used ergogenic substances. In the past 10 years, the amount of data regarding these substances from well-designed clinical trials has increased dramatically. Anabolic steroids remain difficult to study because of their known harmful side effects. The vast amount of data on creatine and exercise performance does not support the dramatic claims of muscle building and power development by manufacturers. Androstenedione has been popularized by high-profile sports stars, but initial studies cast doubt about its performance-enhancing potential. The physician requires factual information about these substances to be able to counsel youth about their effects. PMID- 11862890 TI - Pelvic inflammatory disease in adolescents: a review. AB - Pelvic inflammatory disease is relatively common in sexually active adolescents. Most cases are only mildly symptomatic, and criteria for diagnosis should not be stringent. Although chlamydial or gonorrheal infections or both are common in pelvic inflammatory disease, other aerobic and anaerobic organisms are often also present and may be the only causative agents. Commonly used initial therapy, e.g., ceftriaxone and doxycycline, treats some but not all of these organisms. Patients should be seen within 48 to 72 hours after initiating therapy; lack of improvement suggests noncompliance with antibiotic therapy, the need for broader spectrum antibiotic coverage, a tubo-ovarian abscess, or a mistake in diagnosis. Important preventive measures include screening of sexually active, asymptomatic adolescents for gonorrheal and chlamydial infections, not only in clinics for sexually transmitted diseases and family planning but also in primary care settings. Urine tests that amplify chlamydial and gonococcal nucleic acid are noninvasive and very accurate. PMID- 11862892 TI - Screening, diagnosis, and management of non-insulin-dependent diabetes mellitus in adolescents. AB - Non-insulin-dependent diabetes mellitus (NIDDM), type 2 diabetes mellitus, was once considered rare in children and adolescents. This is no longer true as NIDDM now accounts for 5% to 45% of new cases of diabetes mellitus in the pediatric age group. Most commonly, this disease is seen in obese children with a family history of NIDDM, and particularly in African American, Native American, and Hispanic children. Acanthosis nigricans and hyperandrogenism are sometimes seen. Pediatric health care providers should be aware of the factors that contribute to the development of NIDDM in children as well as the approaches to diagnosis and treatment. PMID- 11862893 TI - The transition into adulthood for children and youth with special health care needs. AB - With advances in medical care, children with chronic medical conditions are living longer lives. They, their families, and their health care professionals are faced with new challenges: transition from pediatric care into adult medical care and meeting the vocational, educational, and other social needs to make this transition of children and youth with special health care needs successful. This article identifies problems and proposed solutions to start to make this transition more successful, including the results of working meetings of representatives from Texas, Oklahoma, New Mexico, and Arkansas. PMID- 11862894 TI - The governor's veto. PMID- 11862895 TI - Telemedicine not needed. PMID- 11862896 TI - [Ethics: from contemplation to concrete evaluation of medical practice]. PMID- 11862897 TI - [The French Journal of Stomatology and Maxillofacial Surgery: what is the future?]. AB - A scientific journal reflects the dynamics of a specialty and should allow wide diffusion of clinical as well as fundamental research findings in order for all practitioners to keep up with changes in their specialty. These objectives are totally different from a journal designed to meet teaching objectives where general reviews, technical notes or clinical cases are in the forefront. The purpose of this work was to determine the current role of the Revue de Stomatologie et de Chirurgie Maxillo-faciale in an attempt to determine future perspectives. MATERIAL AND METHODS: A Medline research was conducted for the period 1998-2000 to ascertain what French papers in the field of stomatology and maxillo-facial surgery had been published in indexed international reviews devoted to the specialty or related areas. The results obtained were compared with articles submitted for publication in the Revue de Stomatologie et de Chirurgie Maxillo-Faciale in 1999. RESULTS: Nineteen French papers were published in indexed international journals. These papers included 10 original articles, 8 cases reports and one technical note. The 10 original articles issued from 8 different centers and the 19 papers from 12 centers. During 1999, 96 papers were submitted to the Revue de Stomatologie et de Chirurgie Maxillo-Faciale for publication. Before acceptation, these papers included original articles (25%) case reports (45%) and technical notes or reviews (30%). In all, 18 original articles issuing from 10 different centers, the same centers that had published in the indexed international journals, were published. DISCUSSION: Considering that the substance of a scientific review lies in the abundance of its original articles, whether the work reports fundamental or clinical research, but more particularly clinical research for a surgery review, one is obliged to observe that the volume of French publications produced annually is insufficient to fullfil the objectives of a truly scientific journal. The 45% submission rate of case reports is in line with this observation. The number of international publications clearly does not explain this situation since globally, over a 2.5 year period, the number of original articles published was to order of one-third of the number published in the Revue de Stomatologie et de Chirurgie Maxillo Faciale in the single year of 1999, and since the teams involved were the same. PMID- 11862898 TI - [Should the lingual nerve be protected during germectomy? A prospective study apropos of 300 procedures]. AB - PURPOSE: Lingual nerve damage is one of the main complications of mandibular third molar removal. In literature, the series published are heterogeneous so we have decided to study the importance of the lingual protection only during germectomies (wisdom teeth without root formation). METHODS: We conducted a randomized prospective study in 300 germectomies, operated by the same skilled surgeon. These 300 third molar removals have been divided in two groups, one with lingual nerve protection and one without. RESULTS: Lingual nerve injuries following germectomies, with or without lingual protection was 0%. CONCLUSION: In germectomies, one can avoid lingual protection. PMID- 11862900 TI - [Early implant treatment of a child with anhidrotic ectodermal dysplasia. Apropos of a case]. AB - The purpose of this article is to report the clinical course and follow-up of a child with ectodermal dysplasia who was treated with implants surgery very early. Different possibilities for prosthetic restoration in the anodontic child are reviewed. Tolerance was excellent. Good cover of the implant was achieved at four years. We propose early implantation reconstruction surgery for these exceptional cases. A prospective multicentric study of this condition would be useful. PMID- 11862899 TI - [Comparison of the cephalometric characteristics of snoring patients and apneic patients as a function of the degree of obesity. Apropos of 162 cases]. AB - BACKGROUND: The purpose of our study was to compare cephalometric analysis of craniofacial features in normal weight or obese subjects who are habitual snorers or apneic. We conducted a retrospective comparison of their clinical and cephalometric features by degree of obesity. PATIENTS AND METHODS: One hundred and sixty two male subjects with obstructive sleep apnea syndrome (OSAS) diagnosed by conventional polysomnography were included in the study. Patients were divided into four groups according to their body mass index (BMI) and their apnea/hypopnea index (AHI: Group 1 = normal-weight snorers (34 patients), Group 2 = normal-weight apneic subjects (40 patients), Group 3 = obese snorers (20 patients), Group 4 = obese apneic subjects (68 patients). Lateral cephalometry was performed in all patients. Intergroup comparisons (2/4, 1/2, 3/4) were made using 32 parameters to study the influence of the size of bone structures, their relationships, and size of the upper airways. RESULTS: The four groups were comparable for age. AHI was higher for group 4 (obese apneic) compared with group 1 (normal-weight snorers). Compared with group 3 (obese snorers), group 1 (normal weight snorers) had a retroposition of the mandible (smaller SNB and ANB angle), an accentuated facial divergence and a narrower pharyngeal space at the hyoid bone level. Compared with group 1 (normal-weight snorers), group 2 (normal-weight apneic) had a narrower pharyngeal space at different levels. Compared with group 3 (obese snorers), group 4 (obese apneic) had a lower hyoid bone evaluated with different cephalometric variables. CONCLUSION: This study mainly shows that apneic patients exhibit craniofacial differences when divided into two groups according to their body mass index. Our findings are consistent with previous reports and could suggest a dual etiology of OSAS. PMID- 11862901 TI - [Primary osseous leiomyosarcoma of the jaw. Apropos of a case]. AB - Primary leiomyosarcoma of bone is a malignant smooth muscle tumour uncommonly found in the jaws. We report a case of primary leiomyosarcoma of the mandible in a 31-year-old man treated with wide resection and radiotherapy then secondary reconstruction with a fibula free flap. No recurrence or distant metastasis has developed four years after the initial surgery. We review the literature on primary leiomyosarcoma of the jaws. PMID- 11862902 TI - [Orthodontic implications in the correction of the transverse dimension in orthognathic surgery]. AB - Transverse skeletal discrepancies are treated in skeletally mature patients with an association of orthodontics and surgical procedures. The orthodontics procedures used depend on the type of surgical maxillary expansion performed. Orthodontics procedures performed before surgery are designed around the analysis of frontal asymmetry. Indications and proper treatment strategy for surgically assisted maxillary expansion, or for Lefort I osteotomy depend on the magnitude of the transverse deficiency. The orthodontics implications of facial asymmetry remains one of the specific points requiring special care before and after surgery. Long length orthodontics stabilization is required after surgically assisted rapid maxillary expansion to limit transversal relapse by use of a rigid fixation or another auxiliary stabilizing arch wire. PMID- 11862903 TI - [Rare earth magnets in conjunction with fixed orthodontics. An "attractive" solution for the positioning of impacted teeth]. AB - Two rare earth permanent magnets are used to align impacted teeth: one is bonded on the crown of the ectopic tooth, the other, intraoral, guides the impacted tooth to the desired place by its attractive power. The intraoral magnet is fixed to an edgewise arch. The adjustments allowed by fixed appliance create better final tooth position than with removable appliance. Our experience depicts an improvement of stability in results and a diminution of periodontal breakdown. PMID- 11862904 TI - Facts are missing. PMID- 11862905 TI - Hearing clearly. PMID- 11862906 TI - Setting the record straight. PMID- 11862907 TI - For the good of the profession. PMID- 11862908 TI - NYSDA prepares for February observance of children's dental health month. PMID- 11862909 TI - To compute or not to compute. That is the question. PMID- 11862910 TI - Walk with a pen, or fly with a computer. Can electronic dentistry work for you? AB - In the last several years, dentistry has crossed over into the new frontier of electronic dentistry. It has embraced such developments as computer programs for producing digital radiographs and photographs, as well as digital programs that enhance these images, store and organize them into a retrievable "chart-like" fashion, and transmit them via the Internet. In Europe, I saw patients with an electronic "health card." This credit card-sized CD can carry all the information on a patient's written charts, results of laboratory tests, radiographic/imaging information and more. It is expected that the mobile phone will be an alternate vehicle for patient records, and that these records will be accessed with a password security system. This will allow patients to carry their records from location to location. Certainly, the dental implications of such seemingly advanced processes are evident. The expression, "The future is now," was never truer. PMID- 11862911 TI - The benefits of a paperless orthodontic practice. AB - After nearly 17 years of incorporating computer technology into our orthodontic practice, we have achieved a "paperless" orthodontic practice. The total integration of computer technology into each facet of our practice has allowed for increased efficiency, simplified clinical activities, improved communications between both patients and professional colleagues and, ultimately, the growth of our practice. This article reviews our current computerized practice management system and outlines the many advantages we have found using this technology in our practice. PMID- 11862912 TI - Marketing your dental practice using microsoft powerpoint. AB - Educating the parents of my patients about the techniques and specialized care pediatric dentists provide has always been a challenge. When I began my dental practice in 1974, the materials and audiovisual programs that were available for parental and patient education were of poor quality and often did not reflect my office philosophy. The multimedia material for pediatric dentistry was limited in scope and of little value in conveying to the parents and patients pediatric restorative procedures, causes of dental disease, caries prevention and patient management. Effective communication to the public can be divided into two areas: in-office marketing; and outside advertising. The primary focus of this discussion will be on in-house marketing of your dental practice using Microsoft PowerPoint. PMID- 11862913 TI - The dental digital radiograph. AB - A discussion of digital radiographs. Shows how the digital radiograph is formed and its many uses. Lists and explains the advantages, that is, it is instantaneous, fewer problems with quality, and can mesh with the computer in the office. The disadvantages are described as well and may include cost, medical liability and storage space. Lastly, it points out the possible future developments that may take place in dentistry with use of this technology. PMID- 11862914 TI - Innocent in the eyes of the law. PMID- 11862915 TI - Orthodontic treatment standards in a public group practice in Sweden. AB - The aim was to assess the orthodontic treatment service provided by 6 orthodontists in a group practice in Malmo. One hundred cases were randomly selected from the model store. The Index of Complexity, Outcome and Need (ICON) was used to assess the need, complexity of the problem, outcome, the degree of improvement and whether the completed case was acceptable or not. The reliability of the examiner using the ICON was assessed using Root Mean Square. Logistic regression analysis was employed to explore the variables related to acceptability of the finish and duration of treatment. The younger the patient at the start of treatment the lower initial ICON score, with short treatment duration were associated with an acceptable finish. Three out of 100 cases were deemed as not requiring orthodontic treatment and 36 cases were classified as very difficult to treat. Nevertheless, 71 cases out of the 100 exhibited acceptable finishes with 27 indicating substantial or great improvement. 6 cases finished treatment with ICON scores greater than 43 indicating need for orthodontic treatment. The treatment on average took 22 months. An objective appraisal of the quality of orthodontic care in a group practice in Malmo has been undertaken. Seventy-one cases were completed with acceptable occlusions. The Index of Complexity, Outcome and Need appears to be a valuable tool to assess the multiple facets of orthodontic provision. PMID- 11862916 TI - Factors associated with snuffing habits among ice-hockey-playing boys. AB - The aim of the present investigation was to study differences in background factors between ice-hockey-playing boys who used snuff, who had tried using snuff, and who had never used snuff. The background factors studied were the socio-economic conditions of the boys' parents; the tobacco habits of the boys' parents, siblings, and friends; the boys' choice of theoretical or practical upper secondary school programme; knowledge of the harmful effects of tobacco as well as participation in a sport other than ice hockey. Data were collected usinG a questionnaire. Of 249 boys in the age group 12-19 years who participated in the study, 13% used snuff, 34% had tried using snuff, and 53% had never used snuff. The factors knowledge of the harmful effects of tobacco and choice of theoretical or practical upper secondary school programme had no significant association with the boys' snuffing habits. This was also true for the parents' tobacco habits unless their habits were analysed separate from each other, where "mother smoked" showed an association with boys who used snuff. The study also showed a significant difference between boys who used snuff and boys who had never used snuff and between boys who had tried using snuff and boys who had never used snuff; boys who used snuff tended to have siblings who used snuff. Of boys who used snuff, 77% reported that their friends' use of snuff had influenced them to start using snuff. A significant difference could also be shown between boys who used snuff, had tried using snuff and never-users depending on whether the boys participated in a sport other than ice hockey. Snuff usage was not as widespread among boys who participated in other sport activities. This was verified by the multivariate logistic regression analysis where "taking part in other sports than ice hockey" was the explanatory variable that showed the strongest association with the dependent variable. An active participation in ice hockey and the environment, in which this sport is practised, would consequently be a strong influencing factor to start using snuff. PMID- 11862917 TI - External apical root resorption as a consequence of orthodontic treatment. PMID- 11862918 TI - A clinical rationale for using guided tissue regenerative (GTR) therapy. PMID- 11862919 TI - Assessing salivary gland hypofunction--Part One. PMID- 11862921 TI - Removable ovate pontic for peri-implant architecture preservation during immediate implant placement. PMID- 11862920 TI - Predictable peri-implant gingival aesthetics: surgical and prosthodontic rationales. AB - Bone resorption following maxillary anterior tooth extraction is common and often compromises gingival tissue levels for the implant restoration. The creation of predictable peri-implant aesthetics requires proper understanding and preservation of the osseous and gingival tissue surrounding the failing tooth. This article addresses the various factors that affect the predictability of peri implant gingival aesthetics in anterior single-tooth replacement. The surgical and prosthodontic procedures for maintenance of peri-implant aesthetics are also presented using an ovate pontic tooth on a removable partial denture following immediate extraction and implant placement. PMID- 11862922 TI - Shade selection techniques: Part Three--principles for stratification. PMID- 11862923 TI - Bone grafts and growth and differentiation factors for regenerative therapy: a review. AB - Guided bone regeneration, tissue grafts, regenerative barrier membranes, and bone substitute materials have been used to restore inadequate hard and soft tissue structures to make them conducive to proper implant placement. Polypeptide growth and development factors (GDFs) have successfully been applied exogenously to periodontal defects to attract preosteoblasts to the site and accelerate their proliferation to stimulate angiogenesis. This article provides an overview of current modalities for restoring lost bone and soft tissue during the treatment of periodontal disease. PMID- 11862924 TI - The electronic clinical patient record. PMID- 11862925 TI - Nonsimultaneous tooth contact in combined implant and natural tooth occlusal schemes. PMID- 11862926 TI - Instrumentation for effective suturing techniques: devices for successful closure. PMID- 11862927 TI - Three-stage approach to aesthetic implant restoration: emergence profile concept. AB - Proper treatment planning and precise evaluation of various parameters (e.g., bone volume, soft tissues, dental anatomy, surgical and prosthetic components) are crucial for an aesthetic implant restoration. The three-stage approach of the emergence profile concept guides the selection of implant, healing abutment, and provisional prosthesis. Adaptation of the implant, provisional prosthesis, and crown restoration stages and their harmonious integration with the soft tissues enable the development of an optimal aesthetic result. This article demonstrates the incorporation of the emergence profile concept for aesthetic implant placement. PMID- 11862928 TI - New technologies. PMID- 11862929 TI - Increase your net on the net. PMID- 11862930 TI - Incorporating high technology for maximum patient comfort. PMID- 11862931 TI - New technologies: their implementation and the future. PMID- 11862932 TI - Office design and technology placement: fundamental foundations. AB - Build technology infrastructure on a solid foundation. Outline the technology vision, determine the budget, research products and vendors, plan a practical installation and support your investment with training. Reap the rewards of today's dental technologies by establishing your practice as high technology yet soft touch. PMID- 11862933 TI - Adjunctive Orchestrated Orthodontic Therapy: an emerging trend in cosmetic dentistry. PMID- 11862934 TI - Three-dimensional cephalometry. PMID- 11862935 TI - The science and mathematics of Dental AART. PMID- 11862936 TI - Clinical significance of Dental AART technology: three-dimensional, volumetric, anatomically accurate individual patient capture. PMID- 11862938 TI - Academy of laser dentistry certification programme 2002. PMID- 11862939 TI - A preliminary study on toothpaste efficacy in reducing levels of S. mutans in dental plaque. PMID- 11862940 TI - The impact of age on tooth colour. PMID- 11862941 TI - Calpol grows up with latest innovation in children's medicine. PMID- 11862942 TI - The ethics of suicide prevention. PMID- 11862943 TI - [The meaning of the term "normal"]. PMID- 11862944 TI - New DOE policy: protection of human subjects. PMID- 11862945 TI - Official 2000-2001 directory of members. PMID- 11862946 TI - Esa1, an Arabidopsis mutant with enhanced susceptibility to a range of necrotrophic fungal pathogens, shows a distorted induction of defense responses by reactive oxygen generating compounds. AB - An Arabidopsis thaliana mutant, esa1, that shows enhanced susceptibility to the necrotrophic pathogens Alternaria brassicicola, Botrytis cinerea and Plectosphaerella cucumerina, but has wild-type levels of resistance to the biotrophic pathogens Pseudomonas syringae pv. tomato and Peronospora parasitica. The enhanced susceptibility towards necrotrophic pathogens correlated with a delayed induction of phytoalexin accumulation and delayed induction of the plant defensin gene PDF1.2 upon inoculation with pathogens. Two reactive oxygen generating compounds, paraquat and acifluorfen, were found to cause induction of both phytoalexin accumulation and PDF1.2 expression in wild-type plants, but this induction was almost completely abolished in esa1. This finding suggests that esa1 may somehow be involved in transduction of signals generated by reactive oxygen species. PMID- 11862947 TI - Lateral root formation is blocked by a gain-of-function mutation in the SOLITARY ROOT/IAA14 gene of Arabidopsis. AB - Lateral root development is a post-embryonic organogenesis event that gives rise to most of the underground parts of higher plants. Auxin promotes lateral root formation, but the molecular mechanisms involved are still unknown. We have isolated a novel Arabidopsis mutant, solitary-root (slr), which has reduced sensitivity to auxin. This dominant slr-1 mutant completely lacks lateral roots, and this phenotype cannot be rescued by the application of exogenous auxin. Analysis with cell-cycle and cell-differentiation markers revealed that the slr-1 mutation blocks cell divisions of pericycle cells in lateral root initiation. The slr-1 mutant is also defective in root hair formation and in the gravitropic responses of its roots and hypocotyls. Map-based positional cloning and isolation of an intragenic suppressor mutant revealed that SLR encodes IAA14, a member of the Aux/IAA protein family. Green fluorescent protein-tagged mutant IAA14 protein was localized in the nucleus, and the gain-of-function slr-1/iaa14 mutation decreased auxin-inducible BA-GUS gene expression in the root, suggesting that SLR/IAA14 acts as a transcriptional repressor. These observations indicate that SLR/IAA14 is a key regulator in auxin-regulated growth and development, particularly in lateral root formation. PMID- 11862949 TI - 3D gold in situ labelling in the EM. AB - We have developed a novel pre-embedding in situ hybridization labelling method for electron microscopy which has given much greater sensitivity and higher labelling levels than have been achieved previously, together with good ultrastructural preservation. Vibratome sections of plant tissue were labelled throughout their thickness with 1 nm gold antibodies and then silver enhanced, embedded in resin and sectioned for electron microscopy. Because the labelling extends throughout the depth of the specimen, this method permits the study of the 3D arrangement of the labelling at the electron microscope level by either stereo-pair recording, tomographic reconstruction or 3D reconstruction from serial sections. In this paper we describe the application of this method to study the organization of rDNA in pea root tissue. PMID- 11862948 TI - An ankyrin repeat-containing protein plays a role in both disease resistance and antioxidation metabolism. AB - The Arabidopsis ankyrin repeat-containing protein AKR2 was identified as a GF14(lambda)-interacting protein in a yeast two-hybrid screening (GF14(lambda) is a 14-3-3 protein). Reduced expression of AKR2 by using the antisense technique results in small necrotic areas in leaves accompanied by higher production of H2O2, similar to the hypersensitive response to pathogen infection in plant disease resistance. Transcripts of genes encoding pathogen-induced protein PR-1 (pathogen-related protein 1) and stress-responsive protein GST6 (glutathione S transferase 6) are increased in antisense plants. The resistance to a bacterial pathogen infection was also increased by at least 10-fold in antisense plants. AKR2 also interacts with another GF14(lambda)-interacting protein, the ascorbate peroxidase 3 that scavenges H2O2 in plant cells. These data suggest that AKR2 may be a negative regulator of PR-1 expression, and is probably involved in the regulation of antioxidation metabolism that is shared by both disease resistance and stress responses. PMID- 11862950 TI - Registration artifacts in international comparisons of infant mortality. AB - Large differences in infant mortality are reported among and within industrialised countries. We hypothesised that these differences are at least partly the result of intercountry differences in registration of infants near the borderline of viability (<750 g birthweight) and/or their classification as stillbirths vs. live births. We used the database of the International Collaborative Effort (ICE) on Perinatal and Infant Mortality to compare infant mortality rates and registration practices in Norway (n = 112484), Sweden (n = 215 908), Israeli Jews (n = 148123), Israeli non-Jews (n = 52 606), US Whites (n = 6 074 222) and US Blacks (n = 1328332). To avoid confounding by strong secular trends in these outcomes, we restricted our analysis to 1987-88, the most recent years for which data are available in the ICE database for all six groups. Compared with Norway (with an infant mortality rate of 8.5 per 1000), the crude relative risks [95% confidence intervals] were 0.75 [0.69,0.81] in Sweden, 0.97 [0.90,1.06] in Israeli Jews, 1.98 [1.81,2.17] in Israeli non-Jews, 0.95 [0.89,1.01] in US Whites and 2.05 [1.95,2.19] in US Blacks. For borderline-viable infants, fetal deaths varied twofold as a proportion of perinatal deaths, with Norway reporting the highest (83.9% for births <500 g and 61.8% for births 500 749 g) and US Blacks the lowest (40.3% and 37.6% respectively) proportions. Reported proportions of live births <500 g varied 50-fold from 0.6 and 0.7 per 10000 in Sweden and Israeli Jews and non-Jews to 9.1 and 33.8 per 10000 in US Whites and Blacks respectively. Reported proportions 500-749 g varied sevenfold from 7.5 per 10000 in Sweden to 16.2 and 55.4 in US Whites and Blacks respectively. After eliminating births <750 g, the relative risks (again with Norway as the reference) of infant mortality changed drastically for US Whites and Blacks: 0.82 [0.76,0.87] and 1.42 [1.33,1.53] respectively. The huge disparities in the ratio of fetal to infant deaths <750 g and in the proportion of live births <750 g among these developed countries probably result from differences in birth and death registration practices. International comparisons and rankings of infant mortality should be interpreted with caution. PMID- 11862951 TI - Reducing the caesarean delivery risk in elective inductions of labour: a decision analysis. AB - To determine whether the vaginal delivery rate is increased in nulliparous women who have positive fetal fibronectin (fFN) testing before elective induction of labour, we performed a decision analysis that tested three options: (1) spontaneous labour; (2) testing nulliparous candidates for elective induction of labour at 39 weeks gestation with fFN and inducing labour if fFN positive (women who are fFN negative are managed expectantly); (3) elective induction of labour for women who are at least 39 weeks. We found that spontaneous labour had a vaginal delivery rate of 90, elective induction 79 and fFN screening 83. At baseline, a mother must be willing to take an additional 7 risk of caesarean delivery to warrant fFN testing or an additional 11 risk of caesarean delivery to warrant elective induction. We conclude that spontaneous labour has the highest vaginal delivery rates. An fFN test in a nulliparous woman may help to raise her likelihood of a vaginal delivery in an elective induction. PMID- 11862952 TI - Saphenoperitoneal shunts for patients with intractable ascites associated with chronic liver disease (Br J Surg 1999;86:882-5). PMID- 11862953 TI - Microcirculatory effects of experimental acute limb ischaemia-reperfusion (Br J Surg 2001;88:816-24). PMID- 11862954 TI - Microcirculatory effects of experimental acute limb ischaemia-perfusion (Br J Surg 2001;88:816-24). PMID- 11862955 TI - Prospective study of primary anastomosis following sigmoid resection for suspected acute complicated diverticular disease (Br J Surgery 2001;88:693-7). PMID- 11862957 TI - Use of minilaparotomy in the treatment of colonic cancer (Br J Surg 2001;88:831 6). PMID- 11862956 TI - Use of minilaparotomy in the treatment of colonic cancer (Br J Surg 2001;88:831 6). PMID- 11862958 TI - JAMA patient page. Postpartum depression. PMID- 11862959 TI - From the Centers for Disease Control and Prevention. Updated recommendations on the use of pneumococcal conjugate vaccine in a setting of vaccine shortage- Advisory Committee on Immunization Practices. PMID- 11862960 TI - From the Centers for Disease Control and Prevention. Respiratory syncytial virus activity--United States, 2000-01 season. PMID- 11862961 TI - From the Centers for Disease Control and Prevention. Rapid assessment of injuries among survivors of the terrorist attack on the World Trade Center--New York City, September 2001. PMID- 11862962 TI - From the Centers for Disease Control and Prevention. Fibrosing skin condition among patients with renal disease--United States and Europe, 1997-2002. PMID- 11862963 TI - JAMA patient page. Palliative care. PMID- 11862964 TI - The role of small airways in lung disease. AB - The small airways constitute one of the least understood areas of the lungs. They play a role in many lung diseases, and small airway pathology results in significant morbidity New approaches to their evaluation may provide insights into this major area of lung disease. Asthma is well recognized as a disease of both large and small airways. Physiological and pathological evidence, from techniques such as post-mortem tissue histological analysis, induced sputum and transbronchial biopsies, has reinforced the concept of the involvement of the entire bronchial tree n the inflammatory process in asthma, In addition to describing the airway pathology in asthma, th s review focuses on the pathogenesis and role of small airway obstruction n other diseases, including chronic obstructive pulmonary disease (COPD), idiopathic pulmonary fibrosis (IPF), sarcoidosis and obliterative bronchiolitis (OB). COPD is characterized by the presence of airflow obstruction resulting from lesions in the small airways. In addition, features compatible with small airways disease are common in IPF, sarcoidosis and OB. Recent advances in pulmonary imaging, such as high-resolution computed tomography (HRCT) and magnetic resonance imaging (MRI) with hyperpolarized 3He, have allowed non-invasive reproducible measurements of structure-function relationships to be made for the small airways. These techniques have great potential for diagnosing changes in small airway function and for assessing responses to treatment. New insights into the contribution of small airways to a range of lung diseases may lead to the development of therapies targeted at this part of the bronchial anatomy. PMID- 11862965 TI - The inflammatory marker serum eosinophil cationic protein (ECP) compared with PEF as a tool to decide inhaled corticosteroid dose in asthmatic patients. AB - The objective of this study was to compare the inflammatory marker eosinophil cationic protein (ECP) with peak expiratory flow (PEF) in determining the therapeutic needs of inhaled corticosteroids in asthma patients assessed as asthma symptoms. A randomized, single-blind study over 6 months was performed at six specialist centres in Europe. In total, 164 adult patients with moderate to severe symptomatic asthma and regular use of inhaled corticosteroids were included. After a run-in period of 2 weeks patients were randomly allocated to the ECP or the PEF monitoring group. The dose of inhaled cort costeroids was adjusted every fourth week based on the current serum ECP value or pre bronchodilator morning PEF values as surrogate markers of therapeutic needs. At the end of the study there were no statistically significant differences in the mean daily symptom score or the percentage of symptom-free days between the two groups. The mean daily dose of inhaled corticosteroids was similar in the two groups at the start of the study but the algorithms used to adjust the dose of inhaled corticosteroids resulted in an increased use of inhaled corticosteroids in both groups. The mean daily dose of inhaled corticosteroids over the whole study period was significantly lower in the ECP group compared withthe PEF group (1246 vs. 1667 microg, P = 0.026). In the ECP group, forced expiratory volume in I sec (FEV)% predicted was lower at the end ofthe study compared with the begining (92% vs. 87%, P = 0 .0009), although there was no significant difference between the two groups. None of the used algorithms for ECP and PEF led to improvement in symptom scores, in spite of increased doses of inhaled corticosteroids. In this respect, both methods were equivalent and insufficient. Recommendations suggesting lung function tests in current guidelines may be difficult to translate into clinical practice, however, a combination of inflammatory markers, lung function and symptoms may still improve asthma control. PMID- 11862966 TI - Splanchnic and mammary nitrogen metabolism by dairy cows fed dry-rolled or steam flaked sorghum grain. AB - Objectives were to determine net release or uptake of alpha-amino N, ammonia N, and urea N across portal-drained viscera, liver, splanchnic, and mammary tissues of lactating Holstein cows (n = 8, 86 +/- 8 d in milk) fed alfalfa hay-based total mixed rations containing 40% dry-rolled or steam-flaked sorghum grain. The total mixed rations were offered at 12-h intervals in a crossover design. Blood samples were obtained from indwelling catheters in the portal, hepatic, and mammary veins and mesenteric or costoabdominal arteries, every 2 h for each cow and diet. Steam-flaking increased in vitro rate of starch hydrolysis compared with dry-rolled sorghum (66 vs. 25%). Diet did not alter dry matter intake (18.2 +/- 0.3 kg). Daily milk yield (27.6 +/- 0.8 kg), efficiency of production, and most milk components did not differ between diets, but fat yield was reduced (0.86 vs. 0.91 kg/d) by steam-flaked sorghum, and lactose concentration was increased (4.99 vs. 4.82%). Blood flows in portal and hepatic veins did not differ between diets. Steam-flaking tended to increase urea N cycling to the gut (162 vs. 95 g/d) compared with dry-rolling of sorghum, whereas net absorption of ammonia N and alpha-amino N across portal-drained viscera were decreased. Net mammary uptake of a-amino N increased more than 20% (83 vs. 67 g/d), resulting in a higher mammary extraction ratio (15 vs. 11%) for steam-flaked versus dry-rolled sorghum. Flaking of sorghum improved the efficiency of postabsorptive N metabolism by increasing urea N cycled to the gut and alpha-amino N uptake by the mammary gland. PMID- 11862967 TI - Splanchnic and mammary nitrogen metabolism by dairy cows fed steam-rolled or steam-flaked corn. AB - Objectives were to determine net release or uptake of a-amino N, ammonia N, and urea N across portal-drained viscera, liver, splanchnic, and mammary tissues of lactating Holstein cows (n = 6; 109 +/- 9 d in milk) fed alfalfa hay-based total mixed rations (TMR) containing 40% steam-rolled or steam-flaked corn grain. The TMR were offered at 12-h intervals in a crossover design. Blood samples were obtained from indwelling catheters in portal, hepatic, and mammary veins and mesenteric or costo abdominal arteries, every 2 h for each cow and diet. Steam flaked compared with steam-rolled corn greatly increased in vitro starch hydrolysis (56 vs. 34%). Daily intake of dry matter (18.4 +/- 0.4 kg/d), starch, N, and net energy for lactation by cows were not altered by processing corn; neither were daily yield of milk (29.1 +/- 0.7 kg/d), fat-corrected milk, nor fat corrected milk per dry matter intake. Steam-flaking tended to increase percent milk protein (2.97 vs. 2.82%; P = 0.07), but not yield, and decrease percent lactose (4.83 vs. 4.94) but not yield. Portal and hepatic blood flows were not affected by diet, nor were net absorption of alpha-amino N and ammonia N. Steam flaking compared with steam-rolling increased urea N cycling to portal-drained viscera (212 vs. 87 g/d) by 140%, estimated mammary uptake and extraction ratio of alpha-amino N. Flaking versus rolling of corn improved N utilization in dairy cows by increasing urea cycling to the gut and uptake of a-amino N by the mammary gland. Higher mammary uptake of alpha-amino N (78 vs. 50 g/d) by dairy cows fed steam-flaked corn tended to increase milk protein content and may explain the previously observed effects of cows fed steam-flaked versus steam-rolled corn. PMID- 11862968 TI - A GnRH/LH surge without subsequent progesterone exposure can induce development of follicular cysts. AB - Our hypothesis was that follicular cysts would develop if cows experienced an estradiol-induced GnRH LH surge in the absence of an ovulatory follicle. Further, we hypothesized that estradiol would fail to induce a subsequent GnRH/LH surge in these cows until they were treated with progesterone. In experiment 1, seven cows were synchronized with a controlled internal drug releasing device (CIDR) for 9 d and each received 500 microg of cloprostenol on d 7. All follicles (> or = 5 mm in diameter) were aspirated at the time of CIDR removal using transvaginal follicular aspiration. Two days after aspiration, cows were treated with 5 mg of estradiol benzoate (EB) to induce a GnRH/LH surge in the absence of an ovulatory sized follicle. All cows had an LH surge following the estradiol treatment and three of seven developed an anovulatory condition that resembled follicular cysts. The four cows that did not develop follicular cysts luteinized remaining cells from one aspirated follicle each. Thus, all cows with a progesterone elevation after the estradiol/GnRH/LH surge had subsequent ovulatory cycles, whereas the absence of progesterone was followed by follicular cysts. After 49 d, the anovulatory cows were induced back to normal cyclicity by insertion of a CIDR for 7 d. In two subsequent experiments, nine of 26 cows were induced to have follicular cysts by follicular aspiration followed by 5 mg of EB. After 26 d of observation, all cystic cows received a second treatment with 5 mg of EB and none of the cows showed an LH surge or ovulation. Cystic cows were untreated (n = 4 controls) or treated for 7 d with a CIDR (n = 5). All cystic cows were subsequently treated for a third time with 5 mg of EB. All CIDR-treated cows had an LH surge and ovulated, whereas none of the control cows had an LH surge or ovulation after the estradiol treatment. Thus, a large follicle anovulatory condition, similar to follicular cysts, can be induced by estradiol induction of a GnRH/LH surge in the absence of subsequent luteinization, and this condition prevents a GnRH/LH surge in response to high doses of estradiol. Progesterone eliminates this condition by reinitiation of GnRH/LH surges in response to estradiol. PMID- 11862969 TI - The effects of social dominance on the production and behavior of grazing dairy cows offered forage supplements. AB - We examined whether separating dominant and subordinate cows affected production or behavior, and the relationships between dominance and production measures, by using grazing cows, some of which were offered forage supplements. In experiment 1, 80 cows were allocated to either pasture only or pasture and a silage supplement offered indoors overnight. Dominance was principally related to body weight, but was also positively correlated with lactation number and negatively correlated with grazing time. In experiment 2, 72 spring-calving cows were classified as dominant or subordinate, on the basis of aggressive interactions, and were divided by two factors; i.e., dominant and subordinate cows grazed together or apart, and with or without a hay supplement, which was offered indoors for 75 min/d. Dominant cows produced more milk than subordinates, probably because of their faster pasture biting rate and hay chewing rate. The hay-supplemented dominant cows produced more milk when kept apart from subordinates, and both dominant and subordinate cows gained more weight and lay down for longer when kept apart. However, when no hay was offered, dominant cows produced more milk and had a faster pasture biting rate when grazed together with subordinates. Cows high in the dominance order were more likely to enter the parlor first, but not to begin grazing first. The results suggest that production can increase when dominant and subordinate cows that are offered forage in a competitive situation are separated, whereas there is no benefit in separating them if they are at pasture all day. PMID- 11862970 TI - Do cows prefer a barn compartment with a grooved or slotted floor? AB - Cow preferences for one of two compartments with different floors were tested in a barn with two symmetrical, connected compartments, each intended for 16 cows. Compartments were identical except for the floor. One floor was grooved longitudinally to the feeding barrier and the other was slotted perpendicular to that barrier. Sixteen pregnant, lactating Holstein-Friesian cows were used. After 14 d of habituation, cow behavior was videotaped for 10 full 24-h d, spread over 22 d. Their daily time budget spent on different activities performed in both compartments was analyzed, as was their relocation and use of the concentrate dispenser in each compartment. On average, cows spent 120 min longer per day in the slotted floor compartment than in the grooved floor compartment, and performed all activities in the daily time budget significantly longer in this compartment. However, testing the preference of individuals revealed that, although five cows significantly preferred the slotted floor compartment, two cows significantly preferred the other compartment, and nine cows had no preference. On the slotted floor, cows ate longer at the feeding barrier and more often drank front-on to the water trough; on the grooved floor they more often drank standing parallel to the water trough. Thus, they seemed to prefer to stand parallel to the slots. Relocation of cows, indicated by index of movement, was not significantly different between the floors. Relocation seemed to be stimulated more by the presence of concentrate dispensers in both compartments than by floor profile. Although one floor type was not clearly preferred over the other, the findings and methods of this study have implications for design and implementation of future preference tests for testing husbandry systems and their components. PMID- 11862972 TI - Seminal plasma hormone concentration after oral application of progesterone. AB - Previous studies have revealed beneficial in vitro effects of progesterone on sperm function. The aim of this pilot study was to prove if orally given micronized progesterone leads to elevations in progesterone and/or 17alpha hydroxyprogesterone levels in seminal plasma, since higher seminal plasma levels of these hormones could possibly have a beneficial effect on sperm function as seen in in vitro investigations. Multiple application of micronized progesterone given over 4 days (daily dose 400 mg) to 6 healthy subjects resulted in elevated seminal plasma levels of progesterone (10.90 +/- 9.02 nmol/l vs. 1.43 +/- 0.56 nmol/l, p = 0.04) and 17alpha-hydroxyprogesterone (3.09 +/- 1.72 nmol/l vs. 1.62 +/- 1.26 nmol/l, p = 0.04) whereas no significant difference could be found in testosterone levels (34.82 +/- 13.00 vs. 30.91 +/- 8.56 nmol/l, p = 0.43). In contrast, androstendione levels in seminal plasma were reduced (2.68 1.28 nmol/l vs. 3.65 +/- 1.36 nmol/l, p = 0.01). Although micronized progesterone is rapidly metabolized, oral application resulted in pronounced elevations of progesterone and 17alpha-hydroxyprogesterone in seminal plasma. Further studies will show if oral application of micronized progesterone can induce beneficial effects on sperm function such as those seen in in vitro investigations. PMID- 11862971 TI - Proteomics and a future generation of plant molecular biologists. AB - Proteomic methods are required for the study of many different aspects of plant function. Important issues in proteomics include the molecular complexity of proteins, given that there are hundreds of thousands of chemically and physically distinct proteins in plants, and the context of protein functions with respect to both genomes and the environment. Available genomic and gene sequences greatly simplify the identification of proteins using improved techniques of mass spectrometry. This improved capability has led to much discussion on proteomes, and some experimentation using proteomic methodologies aimed at modest numbers of proteins. The scale of proteomics is open, for the number of proteins and genes considered at any one time is as dependent on the nature of the scientific question posed as on technical resources and capabilities. We know just enough about plant proteomes to imagine the breathtaking scope of our ignorance. There are tremendous opportunities for new molecular biologists to define the nature of the protein machines that transduce genetic and environmental information, and transform simple energy and matter, to give plants. PMID- 11862973 TI - Comparison of different reduced sampling approaches for the estimation of pharmacokinetic parameters for long half-life drugs in patients with renal or hepatic impairment. AB - OBJECTIVE: To compare 3 different reduced sampling approaches (truncated area, population and Bayesian; sampling schedule till 48 or 72 hours) with the extensive sampling for the estimation of pharmacokinetic parameters for long half life drugs in healthy subjects and in patients with renal or hepatic impairment. METHODS: Two drugs (extensively metabolized or extensively excreted) whose half lives were greater than 30 hours were used in this analysis. Pharmacokinetic parameters such as maximum plasma concentration, clearance and half-life were estimated in healthy subjects and in patients using the above-mentioned 3 reduced sampling approaches and then compared with the extensive sampling. RESULTS: The truncated area method failed to detect the same magnitude of difference in pharmacokinetic parameters between healthy subjects and patient populations that was determined using extensive sampling. On the other hand, the population or Bayesian approach provided the same magnitude of difference in pharmacokinetic parameters between the 2 populations that was observed with extensive sampling. CONCLUSION: This study indicates that the truncated area method may be a less suitable method to accurately characterize the pharmacokinetics of a long half life drug either in healthy subjects or in patients with renal or hepatic impairment compared to a population or Bayesian approach. PMID- 11862974 TI - One-compartment model for amino acids and other biological molecules in peritoneal dialysis. AB - OBJECTIVES: Investigation of the main factors determining the concentration-time course of amino acids and biological molecules in serum and dialysates. METHODS: In a randomized, 3-period crossover study, 11 patients were treated once with each of 3 peritoneal dialysis solutions, 1 containing amino acids and bicarbonate, 1 containing glucose and bicarbonate and 1 containing glucose and lactate. Nineteen amino acids, 3 proteins, 2 metabolites and 2 ions were measured in serum and dialysate. A standard compartment model was fitted to the data. RESULTS: The amino acids differed significantly in their kinetic characteristics (p < 0.001), mainly volume of distribution and elimination rate. Differences in absorption were small compared to the interpatient variation. The average transport rate from serum to dialysate was 0.50-1.14 h(-1), from dialysate to serum 0.33-0.41 h(-1), for elimination from the central compartment 0.35 to 2.27 h(-1), for volume of distribution 0.29 to 0.83 l/kg, for serum protein binding 19 47%, for amount in tissue 82 - 95%, for endogenous metabolic rate 16-151 micromol x kg(-1) x h(-1). The volume of distribution correlated with the R group (polar positive < aliphatic < polar uncharged). For the various proteins, the 2 bicarbonate solutions had higher serum-to-dialysate transport rates than the lactate solution (p = 0.018-0.601). CONCLUSION: The compartment model demonstrated its usefulness. Accordance with literature data for healthy volunteers indicated the validity of the estimates. PMID- 11862975 TI - Effects of acute and chronic alacepril treatment on exercise capacity and hemodynamics in patients with heart failure: a preliminary study. AB - OBJECTIVE: This study determined whether alacepril treatment improves exercise hemodynamics in patients with heart failure. METHODS: Supine bicycle ergometer exercise was performed after administration of placebo and after acute and chronic (12 weeks) alacepril treatment in 4 patients with heart failure. Oxygen uptake (VO2), arterial oxygen saturation (SaO2), and mixed venous oxygen saturation (SvO2) were measured continuously using a pulse oxymeter and a fiber optic catheter. Cardiac index was calculated with Fick's equation. RESULTS: Acute alacepril treatment did not significantly alter the VO2 or hemodynamics. After chronic alacepril treatment, peak VO2 increased (placebo vs chronic alacepril treatment: 17.7 +/- 2.8 vs 21.7 +/- 2.8 ml/min/kg, p < 0.05). Arteriovenous oxygen difference (SaO2 - SvO2) at peak exercise was not altered, however, cardiac index at peak exercise (5.07 0.67 vs 6.35 +/- 0.48 I/min/m2, p = 0.02) increased and stroke volume index at peak exercise (37.3 +/- 3.4 vs 46.5 +/- 1.1 ml/m2, p = 0.07) tended to increase. CONCLUSIONS: Chronic treatment with alacepril improved maximal exercise capacity in patients with heart failure. The increased peak VO2 was primarily due to the increased cardiac index, but not due to the widening of arteriovenous oxygen difference. Therapy-induced increase in stroke volume index may contribute to the increased cardiac index at peak exercise in our patients with heart failure. PMID- 11862976 TI - The revised Declaration of Helsinki--is justice served? AB - The World Medical Association revised the Declaration of Helsinki in October 2000. The Declaration is intended to provide a universal set of principles, which direct the ethical conduct of clinical medical research involving human subjects throughout the world. Previous research ethics codes emphasized the principles of informed consent and beneficence toward research subjects. The revised Helsinki Declaration places a premium not only upon these values but also the principle of distributive justice. The new Declaration demands that scientists see to it that the risks and benefits of scientific inquiry are distributed fairly to those that participate in research and to the communities from where participants are drawn. The authors of the Declaration have sought to establish justice by minimizing the use of placebo controls, insisting that populations from where research subjects are drawn stand to benefit from the research, and by requiring that therapeutic agents be made available to all trial participants long after a trial is completed. This paper argues that the aim of seeking a morejust distribution of the risks and benefits of research may actually be undermined by the tools that the Declaration employs to enforce justice. PMID- 11862977 TI - Orphan drug product regulation--United States. AB - The legislative history of the United States Orphan Drug Act began with rare, unanimous approval by the United States Congress. The Act, mid consequently the Regulations, have evolved since then. The two-stage process of Orphan Drug designation and approval is outlined, as well as the incentives that are offered to commercial companies for their implementation. Orphan Drugs are likely to be over-represented among drugs used under "Treatment" INDs. For patent- and "drug difference" reasons, the benefits under the Orphan Drug Act are especially valuable to those who develop biologics. By any measure, this legislation, which requires only voluntary participation, has been a success; because the human genome is likely to lead to more biologicals than orthodox drugs, this success is likely to continue into the future. But even so, the 18-year experience with Orphan Drugs in the United States has led to some 225 Orphan Product approvals that benefit many millions of patients. PMID- 11862978 TI - Development of a preference-weighted health status classification system in France: the Health Utilities Index 3. AB - The Health Utilities Index is a generic multiattribute preference-based system for assessing health-related quality of life, devised by Torrance et al. It is being used in cost-effectiveness evaluations in North America and in international multicentre studies but was not available in France. Following adaptation of the HUI3 classification in France, the purpose of the reported investigation was to derive French preference weights. This article provides a reminder of the theoretical foundations used to model the multiattribute utility function. Within this framework, a multiattribute multiplicative aggregate utility function was constructed in accordance with the explicitly decomposed approach. The study took place in June 1999 over a sample of 365 persons from the French general population, aged between 20 and 65, and not suffering from any chronic or incapacitating illness. The recruitment procedure was based upon a random selection of individuals, using the phone book. Interviews took place in the homes of the interviewees. The methods of revelation (VAS and SG) were applied by setting the value of the best possible state of the HUI3 a priori at 1, and leaving a choice between two states (worst possible state, death) for 0. The aggregated individuals (person-mean and median) were calculated and the multiplicative utility functions constructed. A comparison of the calculated utilities with the observed ones provides a primary indicator of the validity of the person-mean or median functions constructed. The slight absolute differences obtained between observed and calculated utilities and the low RMSE scores lead us towards a favourable conclusion. PMID- 11862979 TI - Modelling efficiency, screening and preferences within healthcare systems. PMID- 11862980 TI - Identification of differentially expressed transcripts in the human pathogenic fungus Paracoccidioides brasiliensis by differential display. AB - Paracoccidioides brasiliensis is a dimorphic human pathogenic fungus that is the causal agent of paracoccidioidomycosis, a systemic disease that predominantly affects rural communities in South and Central America. Dimorphism is a common characteristic of systemic human pathogenic fungi. Here we describe the use of differential display (DD) to isolate and identify differentially expressed genes of P. brasiliensis, in the two cell types, yeast (Y) and mycelium (M), as well as at different time intervals during temperature-induced M to Y transition. Using two oligo-deoxythymidine-anchored primers combined with 10 arbitrary ones, we were able to detect the presence of at least 20 differentially transcribed cDNA fragments. Some of these fragments were further analysed by reverse-northern blot and northern blot in order to confirm their differential expression. The M32, M51 and M73 cDNA fragments were specific for the mycelial form of P. brasiliensis. Furthermore, we found two cDNA fragments (M-Y1 and M-Y2) that were upregulated during M-Y transition. This method was efficient and useful in the detection of differentially expressed genes in P. brasiliensis. PMID- 11862981 TI - Generalized lymphadenopathy caused by Trichosporon asahii in a patient with Job's syndrome. AB - Of the six species of Trichosporon known to cause human infections, T. asahii is the main agent of invasive trichosporonosis. We describe an unusual case of generalized lymphadenopathy due to T, asahii in a 10-year-old boy with Job's syndrome (markedly elevated IgE with eosinophilia). The diagnosis was based on the presence of blastic conidia and hyphal elements breaking into arthroconidia in biopsied tissue of the cervical lymph node and isolation of the causal agent T, asahii in pure culture. The patient responded initially to amphotericin B therapy, but the infection recurred within 4 weeks and did not respond to therapy of liposomal amphotericin B and 5-fluorocytosine for 10 days. The patient left the hospital against medical advice. PMID- 11862983 TI - Effect of feed withdrawal prior to slaughter on prevalence of gastric ulcers in pigs. AB - OBJECTIVE: To determine whether withdrawing feed from pigs prior to slaughter had any effects on prevalence or severity of gastric ulcers. DESIGN: Split-plot design. ANIMALS: 873 pigs. PROCEDURES: At the finishing barn, pigs were assigned to 30 pens. Feed withdrawal times (0, 12, or 24 hours) were assigned to pens at random, and pigs in each pen were marketed in 3 groups over a period of 4 weeks. The first marketing group consisted of the 10 heaviest pigs in each pen, the second consisted of the next 10 heaviest, and the third consisted of all remaining pigs. Feed was withheld from all pigs in each pen prior to removal of each marketing group. Thus, feed was withheld once, twice, or 3 times for pigs in the first, second, and third marketing groups, respectively. RESULTS: Feed withdrawal time was not significantly associated with ulcer score at the time of slaughter. Ulcer scores and prevalence of chronic damage were higher in the third marketing group, regardless of feed withdrawal time. Prevalence of severe damage, prevalence of chronic damage, and prevalence of esophageal constriction increased as carcass weight decreased. No pigs died of gastric ulceration. CONCLUSION AND CLINICAL RELEVANCE: Results suggest that withdrawal of feed from pigs prior to slaughter does not increase damage to the stomach and that repeated feed withdrawal does not result in fatal gastric ulceration. PMID- 11862984 TI - Effects of testicular biopsy in clinically normal bulls. AB - OBJECTIVE: To determine whether testicular needle biopsy is detrimental to testicular function in clinically normal bulls. DESIGN: Prospective study. ANIMALS: 6 mixed-breed mature bulls. PROCEDURE: A randomly selected testicle from each bull was biopsied with a 14-gauge needle biopsy instrument. Bulls were then evaluated over a 90-day period for changes in scrotal temperature and thermal patterns, ultrasonographic appearance, and quality of spermatozoa. At the end of the 90-day study, bulls were castrated, and testicles were examined grossly and histologically. RESULTS: Changes were detected in scrotal temperatures and thermal patterns and in the breeding soundness examination results during the first 2 weeks of the study. However, there were no long-term changes in semen quality over the course of the experiment. Hyperechoic areas were detected on ultrasonographic examination and corresponded to the areas of penetration by the biopsy instrument. Microscopic lesions that were indicative of testicular dysfunction were not found. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicate that testicular biopsy is a safe procedure in bulls. Testicular biopsy could possibly be used to further examine bulls that have less than satisfactory results for breeding soundness examinations. PMID- 11862985 TI - CD164. PMID- 11862986 TI - Detection of adulterants in olive oil by headspace-mass spectrometry. AB - In the present work, we propose the use of direct coupling of a headspace sampler to a mass spectrometer for the detection of adulterants in olive oil. Samples of olive oils were mixed with different proportions of sunflower oil and olive pomace oil, respectively, and patterns of the volatile compounds in the original and mixed samples were generated. Application of the linear discriminant analysis technique to the data from the signals was sufficient to differentiate the adulterated from the non-adulterated oils and to discriminate the type of adulteration. The results obtained revealed 100% success in classification and close to 100% in prediction. The main advantages of the proposed methodology are the speed of analysis (since no prior sample preparation steps are required), low cost, and the simplicity of the measuring process. PMID- 11862982 TI - Effect of withdrawing feed from swine on meat quality and prevalence of Salmonella colonization at slaughter. AB - OBJECTIVE: To determine whether withholding feed from pigs prior to slaughter had any effects on meat quality, percentage of pigs with Salmonella spp in cecal contents during slaughter, or percentage of pigs with lacerations of the gastrointestinal tract during slaughter. DESIGN: Split-plot design. ANIMALS: 873 pigs. PROCEDURES: At the finishing barn, pigs were assigned to 30 pens. Feed withdrawal times were assigned to pens at random, and pigs in each pen were marketed in 3 groups. The first marketing group consisted of the 10 heaviest pigs in each pen, the second consisted of the next 10 heaviest pigs, and the third consisted of all remaining pigs. RESULTS: Withdrawing feed improved the redness score assigned to the meat but did not have any other significant effects on carcass composition or meat quality. The percentage of pigs with Salmonella spp in the cecal contents decreased from the first (73%) to the second (64%) to the third (52%) marketing group. However, isolation of Salmonella spp from cecal contents was not associated with feed withdrawal time or with pen prevalence of Salmonella shedding during the 2 months prior to slaughter. Feed withdrawal time and marketing group did not have any significant effects on overall prevalence of gastrointestinal tract lacerations. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that withdrawal of feed from pigs prior to slaughter does not increase the prevalence of Salmonella colonization or the risk of carcass contamination associated with gastrointestinal tract lacerations during slaughter but only slightly enhances meat quality. PMID- 11862987 TI - Effect of ionic strength on perfusive flow in capillary electrochromatography columns packed with wide-pore stationary phases. AB - The use of wide-pore stationary phases in capillary electrochromatography has shown exceptional increases in separation efficiency in conjunction with high electroosmotic flow. These effects are due to the perfusive flow mechanism which is primarily controlled by the ionic strength of the mobile phase. Good correlation between calculated values of electrochemical double-layer thickness and efficiency data have also been obtained. Reduced plate height values of <0.5 have been observed with pore sizes of 4000 A. In addition, electroosmotic flow mobility twice that of 3 microm Spherisorb ODS-1 has been obtained. PMID- 11862988 TI - Melatonin reverses neurochemical alterations induced by 6-OHDA in rat striatum. AB - The present work showed that the intrastriatal injection of 6-OHDA significantly decreases DA, DOPAC and HVA levels in that rat brain structure. Although there is also a decrease in 5-HT levels no changes were observed in 5-HIAA levels as compared to controls. On the other hand, melatonin (2, 5, 10 and 25 mg/kg. i.p., daily for 7 days) treatment starting 1 h after 6-OHDA lesions, partially reverses the decreases caused by 6-OHDA lesions on these neurotransmitter levels, and contents were brought to approximately 50% of that observed in the contralateral sides of controls or of melatonin treated group. Melatonin was more efficient at the doses of 5 and 10 mg/kg, i.p., and effects were similar between the lowest and highest doses characteristic of a bell-shaped type of response. The apomorphine-induced rotational behavior (3 mg/kg, i.p.) was blocked by 60, 89, 78 and 47% after the doses of 2, 5, 10 and 25 mg/kg, i.p., respectively. Similarly, in this case the doses of 5 and 10 mg/kg were also more efficient. Melatonin (5 mg/kg) produced an upregulation of D1 receptors associated with a decrease in Kd value. While no change was observed in maximum density of D2 receptors, the Kd value was also decreased. PMID- 11862989 TI - Potential role of potassium channel openers in the treatment of asthma and chronic obstructive pulmonary disease. AB - Airway smooth muscle (ASM) cells express various types of potassium (K+) channels which play a key role in determining the resting membrane potential, a relative electrical stability and the responsiveness to both contractile and relaxant agents. In addition, K+ channels are also involved in modulation of neurotransmitter release from airway nerves. The most important K+ channels identified in airways include large and small Ca2+-activated, delayed-rectifier, and ATP-sensitive channels. These K+ channels are structurally and functionally different, thus playing distinct roles in airway electrophysiology and pharmacology. Many in vitro and in vivo studies, performed in both animals and humans, have shown that K+ channel openers are able to induce hyperpolarization of ASM cells, bronchodilation, suppression of airway hyperresponsiveness (AHR), and inhibition of neural reflexes. Therefore, airway K+ channels represent a suitable pharmacological target for the development of new effective therapeutic options in the treatment of asthma and chronic obstructive pulmonary disease (COPD). PMID- 11862990 TI - Field evidence for density-dependent facilitation amongst Ixodes ricinus ticks feeding on sheep. AB - Engorged Ixodes ricinus nymphs collected from sheep resident in an upland UK field site were significantly lighter than nymphs that engorged on previously tick-naive sheep, indicating that site-resident sheep continually exposed to ticks acquired anti-tick resistance. The weights of engorged nymphs that fed on naturally tick-resistant sheep increased significantly, however, when increasingly high numbers of adult female ticks fed on the sheep during seasonal peaks of tick activity. This relationship was unaffected by variations in nymph weight amongst individual sheep, between seasons and years, and potential effects of sheep infection with Ehrlichia phagocytophila; this suggests that high adult tick infestations may directly inhibit the expression of acquired anti-tick resistance by sheep. The length, width and weight of adult ticks and the scutum length of adult females were linearly related to their weight as an engorged nymph. The mean scutum length of adult female ticks feeding on sheep in the field site was greater than that of adult females obtained from engorged nymphs collected from sheep of the same site. This suggests that larger ticks have a survival advantage and that I. ricinus ticks exhibit density-dependent intraspecific facilitation at high infestation levels with potential consequences for the transmission of tick-borne diseases. PMID- 11862991 TI - Natural Ehrlichia phagocytophila transmission coefficients from sheep 'carriers' to Ixodes ricinus ticks vary with the numbers of feeding ticks. AB - In a longitudinal study in a UK upland site, 38% of adult sheep were detected as infected with the tick-borne bacterium Ehrlichia phagocytophila by PCR of blood samples. Infection prevalence declined significantly with sheep age but varied significantly and non-linearly with the number of adult Ixodes ricinus ticks feeding per sheep. These findings suggested that under conditions of natural repeated tick-borne challenge sheep remain partially susceptible to re infections, but the likelihood of re-infection depended on the numbers of feeding ticks. Transmission efficiency from sheep to immature ticks also varied significantly and non-linearly with the number of adult ticks feeding per sheep: transmission efficiency was almost zero in sheep with low adult tick infestations rising to 30% at certain levels of adult tick infestation. Infection intensity in infected engorged immature ticks also varied with the number of adult ticks feeding per sheep, but neither prevalence nor intensity of infection in engorged ticks were related to sheep blood PCR result. These findings suggest that variation in the numbers of ticks feeding per sheep may influence E. phagocytophila transmission by direct effects on transmission at the tick-host interface. PMID- 11862992 TI - Characterization and cytochemical localization of an ATP diphosphohydrolase from Leishmania amazonensis promastigotes. AB - An ATP diphosphohydrolase was identified in the plasma membranes isolated from promastigote forms of Leishmania amazonensis. Both ATP and ADP were hydrolysed at similar rates by the enzyme. Other nucleotides such as UTP, GTP and CTP were also degraded, revealing a broad substrate specificity. Adding ATP and ADP simultaneously, the amount of hydrolysis achieved was compatible with the presence of a single enzyme. ATPase activity was not affected by addition of vanadate, ouabain, thapsigargin, dicyclohexylcarbodiimide, oligomycin and bafilomycin A, thus excluding involvement of P-, F- and V-type ATPases. The effects of pH in the range 6.5-8.5 were examined using ATP or p-NPP as substrate. At pH 7.4, the phosphatase activity decreased, and did not show a significant contribution to ATP hydrolysis. In addition, the enzyme was not inhibited by levamisole and ammonium molybdate, excluding alkaline phosphatase and nucleotidase activities, respectively. Sodium azide (5-10 mM) caused inhibition of the ATP and ADP hydrolysis in a dose-dependent manner. Calcium was the best activating metal ion for both ATPase and ADPase activities. Ultrastructural cytochemical microscopy showed ATP diphosphohydrolase on the surface and flagellar pocket of the parasite. We have proposed that L. amazonensis ATP diphosphohydrolase may participate in the salvage pathway of nucleosides. PMID- 11862993 TI - Experimental evidence of hybrid breakdown between the two geographical strains of Schistosoma intercalatum. AB - Reciprocal crosses (Schistosoma intercalatum male Zaire x S. intercalatum female Cameroon: S. intercalatum female Zaire x S. intercalatum male Cameroon) were produced in 10 mice by exposing each mouse to 60 male cercariae of one isolate and 60 female cercariae of the other isolate, and vice versa. Hybrid generations originating from the two crosses were established. The infectivity of the F1, F2, F3 and F4 hybrid generations were evaluated after exposing snails individually to 5 miracidia. A comparative histological study of snails infected with F2 and F4 hybrid sporocysts from both crosses was made to assess abnormalities in the intramolluscan development of the hybrids. The worm recovery rate and fecundity were measured by comparing the fitness of the mid-parents with that of the hybrids. S. intercalatum Cameroon was compatible with Bulinus forskalii and incompatible with B. globosus whereas S. intercalatum Zaire was compatible with B. globosus and incompatible with B. forskalii. In the case of S. intercalatum male Cameroon x S. intercalatum female Zaire, hybrid miracidia develops in both B: forskalii and B. globosus in F1, F2 and F3 generations. The infection rate was much lower for B. globosus and F2 and F3 generations produced few cercariae (less than 30 cercariae in overall per snail) and F4 miracidia were only infective to B. forskalii again producing few cercariae. At 40 days post-infection the sporocyst contains masses of acidophilic granules originating from the breakup of pycnotic nuclei. Similarly the F1, F2 and F3 miracidia of the reciprocal cross (S. intercalatum male Zaire x S. intercalatum female Cameroon) exhibited a dual infectivity for B. forskalii and B. globosus, but cercarial productivity was low (less than 30 cercariae overall per snail for F2 and F3 generations). Histological studies demonstrated sporocyst degeneration in snails infected with F4 generation. In the definitive host, the F, generation (both crosses) exhibited hybrid vigour in that the worm return was greater than that of the mid-parent, F2 and F3 generations. No significant difference in fecundity was demonstrated between the parental strains and the F1 and F2 generations, yet egg production of the F3 generation was significantly lower. It is apparent that there is a post zygotic barrier in the crosses of S. intercalatum Zaire and S. intercalatum Cameroon from the F2 generations onwards, and sterility of the F4 hybrid sporocyst supports the concept of the existence of 2 distinct taxa. PMID- 11862994 TI - Two main clusters within Trypanosoma cruzi zymodeme 3 are defined by distinct regions of the ribosomal RNA cistron. AB - Trypanosoma cruzi is currently classified into 2 major phylogenetic lineages, T. cruzi I and II, that correlate with the formerly described zymodeme 1 and 2, respectively. Another isoenzymic group (zymodeme 3-Z3) was also described. In this study, we analysed the genetic diversity among Z3 isolates of the Brazilian Amazon by restriction fragment length polymorphism of the intergenic transcribed spacers (ITSs) of the ribosomal RNA cistron and the size of the divergent domain D7 of the 24Salpha rRNA gene. DNAs from 12 T. cruzi Z3 isolates obtained from humans (2), Panstrongylus geniculatus (1), and Rhodnius brethesi (9) were submitted to PCR amplification of the ITSs plus the 5.8S rDNA. The PCR products were digested with 4 distinct endonucleases and the profiles analysed by a numerical methodology. The phenetic dendrogram revealed a clear dichotomy in the Z3 group, defining 2 groups that were named Z3-A and Z3-B. Dimorphism was also found in the band sizes of the amplified D7 divergent domain of the 24Salpha rDNA, which showed a perfect correlation with the ITSs clustering. The organization of the ribosomal cistron was investigated by Southern blotting and shown to be conserved in the genome of the 2 Z3 groups. This study shows that the rDNA cistron allows the definition of 2 distinct subclusters in Z3 isolates. PMID- 11862996 TI - Secondary traumatization in Dutch couples of World War II survivors. AB - This study examined whether signs of secondary traumatic stress were present in a community sample of couples who experienced World War II. The authors hypothesized that symptoms of posttraumatic stress disorder (PTSD) in either spouse may be predicted not only by his or her own war experiences but also by the war experiences and posttraumatic symptoms of the partner. Approximately 50 years after the end of World War II, 444 couples from a community sample of elderly Dutch citizens answered a questionnaire. A multilevel regression analysis was performed with symptoms of PTSD as the dependent variable. The most important predictors of PTSD symptoms were the number of war events reported by the participant and the current level of PTSD symptoms of his or her spouse. The results lend empirical support to the notion that posttraumatic stress reactions of both members of a couple are not independent from each other. Several explanations of the findings are discussed. PMID- 11862995 TI - Bancroftian filariasis: a 13-year follow-up study of asymptomatic microfilariae carriers and endemic normals in Orissa, India. AB - The natural history of human filarial infections leading to development of disease has been a subject of intense debate. The models proposed so far have largely been based on cross-sectional data on microfilariae (Mf) and disease prevalence in filariasis endemic areas. In an attempt to study the parasitological and clinical consequences of filarial infection in Beldal (Orissa, India), an area endemic for Bancroftian filariasis, cohorts of 59 asymptomatic Mf carriers (AS) and 187 asymptomatic and amicrofilaraemic subjects or 'endemic normals' ('EN'), were followed-up and a fraction (73% and 46% respectively) re-examined after 13 years to monitor (a) Mf prevalence, (b) Mf density, (c) circulating filarial antigen (CFA) and (d) chronic disease manifestations. The Mf prevalence and density were also monitored in Mf carriers after 1 and 4 years. Both Mf prevalence and density decreased progressively in the cohort of Mf carriers over a period of 13 years in Beldal. Only 37% of them continued to be microfilaraemic and the Mf density in these subjects was only 10% of the original level. However, loss of circulating Mf in this cohort did not result in loss of CFA and 95% remained CFA positive regardless of Mf status. About 23% of males in the 'EN' cohort developed hydrocoele while only 5.7% of male Mf carriers, who were not treated with DEC, had developed hydrocoele after 13 years. A cohort of Mf carriers in another area, Jatni, was also examined after 10 years to study the parasitological and clinical outcome. In this area, about 59% of the Mf carriers continued to be microfilaraemic after 10 years. These results reveal that in Mf carriers adult filarial worms persist for several years and that loss of circulating Mf with or without chemotherapy with DEC (single 12 day course) does not influence adult worm survival. The findings have been discussed in the context of 'static' and 'dynamic' models describing the relationship between infection and disease in human filariasis. PMID- 11862997 TI - Effects of netobimin treatment on the glucose and glycogen contents of Echinococcus granulosus cysts from gerbils. PMID- 11862999 TI - European Union plans electronic health card to guarantee treatment. PMID- 11863000 TI - Medical journals can reduce global health inequity, conference told. PMID- 11862998 TI - Detection of fowl adenovirus associated with hydropericardium hepatitis syndrome by a polymerase chain reaction. AB - Hydropericardium hepatitis syndrome (HHS), previously unknown in the broiler industry, is an emerging disease that causes severe hydropericardium. A polymerase chain reaction (PCR) was developed to detect the fowl adenovirus (FAV) associated with HHS. The virus from infected livers was purified, with confirmation by electron microscopy and experimental infection. Methods were developed to isolate the viral DNA from purified virus and infected tissues. Available sequence data on the hexon gene of fowl adenoviruses and other adenoviruses were aligned to determine the conserved and variable regions. Primers were constructed from the alignment data. The amplified fragment consisted of the variable region of the hexon gene flanked by conserved primer sites. Optimum conditions were standardized to achieve the amplification of the desired fragment. As expected, the amplified product was found to be of 0.7 kg size. The nucleotide sequence analysis confirmed the specific nature of the product. Amplification of the specific product could be obtained not only from the DNA isolated from the purified virus but also from the total DNA extracted from infected tissues. The PCR was useful for the detection of FAV associated with HHS. PMID- 11863001 TI - Postoperative starvation after gastrointestinal surgery. Meta-analysis was not appropriate. PMID- 11863002 TI - Postoperative starvation after gastrointestinal surgery. Anaesthetic technique during gastrointestinal surgery has postoperative effects. PMID- 11863003 TI - Postoperative starvation after gastrointestinal surgery. Rest in peace, drip and suck. PMID- 11863004 TI - Cardiotocography v Doppler auscultation. Evidence is not as good as NICE suggests. PMID- 11863005 TI - Cardiotocography v Doppler auscultation. Conclusions do not recognise difference between statistical and clinical significance. PMID- 11863006 TI - Cardiotocography v Doppler auscultation. All unbiased comparative studies should be published. PMID- 11863007 TI - Cardiotocography v Doppler auscultation. Trials are underpowered. PMID- 11863008 TI - Gastro-oesophageal reflux disease in general practice. Utility and acceptability of Infai C13-urea breath test has been shown. PMID- 11863009 TI - Caesarean section rate in England and Wales. Caesarean section rates in Hong Kong. PMID- 11863010 TI - Management of chronic disease. Proactive medicine is better option. PMID- 11863011 TI - Management of chronic disease. Life is a chronic disease. PMID- 11863012 TI - Lariam's legacy. AB - The most-prescribed malaria drug could produce psychiatric side effects in more than one-quarter of all travelers who take it. PMID- 11863013 TI - Greco v. United States: a step forward or backward from Roe v. Wade? PMID- 11863014 TI - Ethical issues in HIV vaccine trials in South Africa. AB - In this review we describe the ethical issues central to local and international debates about HIV vaccine trials. These issues include the physiological and psycho-social risks of trial participation, the preventative interventions to be provided to participants, access to treatment for participants who seroconvert, access to an effective vaccine after the trial, the role of placebo-controlled trials, and obtaining informed consent. PMID- 11863015 TI - Conceptualizing the right to health: a violations approach. PMID- 11863016 TI - Women's health and managed care. PMID- 11863017 TI - Camping trips and family trees: must Tennessee physicians warn their patients' relatives of genetic risks? PMID- 11863018 TI - Testing the Fourth Amendment for infection: mandatory AIDS and HIV testing of criminal defendants at the request of a victim of sexual assault. PMID- 11863019 TI - Genetic discrimination: legislation required to keep genetic secrets. PMID- 11863020 TI - Live through this...physician assisted suicide. PMID- 11863021 TI - Physicians, bound and gagged: federal attempts to combat managed care's use of gag clauses. PMID- 11863022 TI - Sweden's shameful eugenics policies. PMID- 11863023 TI - Solving the insurance/genetic fair/unfair discrimination dilemma in light of the Human Genome Project. PMID- 11863024 TI - Genetics, genetic testing, and the specter of discrimination: a discussion using hypothetical cases. PMID- 11863025 TI - "Unnecessary suffering": the cornerstone of animal protection legislation considered. AB - Causing "unnecessary suffering" has been widely adopted in legislation to define criminal liability in respect of the treatment of animals. This article examines the way in which the term has been interpreted and applied by the courts, and considers its effectiveness in affording animals protection from abuse. PMID- 11863026 TI - The religion clauses and parental health care decisionmaking for children: suggestions for a new approach. PMID- 11863027 TI - Constitutional challenges to bans on "assisted suicide": the view from without and within. PMID- 11863028 TI - "Out, out brief candle": constitutionally prescribed suicide for the terminally ill. PMID- 11863029 TI - Womb for rent: Norplant and the undoing of poor women. PMID- 11863030 TI - Solid state microdosimetry. AB - A review of solid state microdosimetry is presented with an emphasis on silicon based devices. The historical foundations and basics of microdosimetry are briefly provided. Various methods of experimental regional microdosimetry are discussed to facilitate a comparison with the more recent development of silicon microdosimetry. In particular, the performance characteristics of a proportional gas counter and a silicon microdosimeter are compared. Recent improvements in silicon microdosimetry address the issues of requirement specification, non spherical shape, tissue equivalence, sensitive volume definition (charge collection complexity) and low noise requirements which have previously impeded the implementation of silicon-based microdosimetry. A prototype based on silicon on-insulator technology is described along with some example results from clinical high LET radiotherapy facilities. A brief summary of the applications of microdosimetry is included. PMID- 11863031 TI - Radiation dosimetry using three-dimensional optical random access memories. AB - Three-dimensional optical random access memories (3D ORAMs) are a new generation of high-density data storage devices. Binary information is stored and retrieved via a light induced reversible transformation of an ensemble of bistable photochromic molecules embedded in a polymer matrix. This paper describes the application of 3D ORAM materials to radiation dosimetry. It is shown both theoretically and experimentally, that ionizing radiation in the form of heavy charged particles is capable of changing the information originally stored on the ORAM material. The magnitude and spatial distribution of these changes are used as a measure of the absorbed dose, particle type and energy. The effects of exposure on 3D ORAM materials have been investigated for a variety of particle types and energies, including protons, alpha particles and 12C ions. The exposed materials are observed to fluoresce when exposed to laser light. The intensity and the depth of the fluorescence is dependent on the type and energy of the particle to which the materials were exposed. It is shown that these effects can be modeled using Monte Carlo calculations. The model provides a better understanding of the properties of these materials. which should prove useful for developing systems for charged particle and neutron dosimetry/detector applications. PMID- 11863032 TI - Space radiation dosimetry in low-Earth orbit and beyond. AB - Space radiation dosimetry presents one of the greatest challenges in the discipline of radiation protection. This is a result of both the highly complex nature of the radiation fields encountered in low-Earth orbit (LEO) and interplanetary space and of the constraints imposed by spaceflight on instrument design. This paper reviews the sources and composition of the space radiation environment in LEO as well as beyond the Earth's magnetosphere. A review of much of the dosimetric data that have been gathered over the last four decades of human space flight is presented. The different factors affecting the radiation exposures of astronauts and cosmonauts aboard the International Space Station (ISS) are emphasized. Measurements made aboard the Mir Orbital Station have highlighted the importance of both secondary particle production within the structure of spacecraft and the effect of shielding on both crew dose and dose equivalent. Roughly half the dose on ISS is expected to come from trapped protons and half from galactic cosmic rays (GCRs). The dearth of neutron measurements aboard LEO spacecraft and the difficulty inherent in making such measurements have led to large uncertainties in estimates of the neutron contribution to total dose equivalent. Except for a limited number of measurements made aboard the Apollo lunar missions, no crew dosimetry has been conducted beyond the Earth's magnetosphere. At the present time we are forced to rely on model-based estimates of crew dose and dose equivalent when planning for interplanetary missions, such as a mission to Mars. While space crews in LEO are unlikely to exceed the exposure limits recommended by such groups as the NCRP, dose equivalents of the same order as the recommended limits are likely over the course of a human mission to Mars. PMID- 11863033 TI - Review of abortion policy: legality, medicaid funding, and parental involvement, 1967-1994. PMID- 11863034 TI - Understanding the nature of the general factor of intelligence: the role of individual differences in neural plasticity as an explanatory mechanism. AB - The nature of the general factor of intelligence, or g, is examined. This article begins by observing that the finding of a general factor of intelligence appears to be inconsistent with current findings in neuroscience and cognitive science, where specific connections are argued to be critical for different intellectual abilities and the brain is argued to develop these connections in response to environmental stimuli. However, it is then observed that if people differed in neural plasticity, or the ability to adapt their connections to the environment, then those highly developed in one intellectual ability would be highly developed in other intellectual abilities as well. Simulations are then used to confirm that such a pattern would be obtained. Such a model is also shown to account for many other findings in the field of intelligence that are currently unexplained. A critical period for intellectual development is then emphasized. PMID- 11863035 TI - Making trade-offs: a probabilistic and context-sensitive model of choice behavior. AB - The stochastic difference model assumes that decision makers trade normalized attribute value differences when making choices. The model is stochastic, with choice probabilities depending on the normalized difference variable, d, and a decision threshold, delta. The decision threshold indexes a person's sensitivity to attribute value differences and is a free estimated parameter of the model. Depending on the choice context, a person may be more or less sensitive to attribute value differences, and hence delta may be used to measure context effects. With proportional difference used as the normalization, the proportional difference model (PD) was tested with 9 data sets, including published data (e.g., J. L. Myers, M. M. Suydam, & B. Gambino, 1965; A. Tversky, 1969). The model accounted for individual and group data well and described violations of stochastic dominance, independence, and weak and strong stochastic transitivity. PMID- 11863036 TI - Frames of reference in unilateral neglect and visual perception: a computational perspective. AB - Neurological patients with unilateral neglect fail to orient and respond to stimuli on one side, typically the left. A key research issue is whether neglect is exhibited with respect to the left side of the viewer or of objects. When deficits in attentional allocation depend not merely on an object's location with respect to the viewer but on the object's intrinsic extent, shape, or movement, researchers have inferred that attention must be operating in an object-based frame of reference. Simulations of a view-based connectionist model of spatial attention prove that this inference is not logically necessary: Object-based attentional effects can be obtained without object-based frames. The model thus explains away troublesome phenomena for view-based theories of object recognition. PMID- 11863037 TI - Pattern and process in the evolution of learning. AB - A century after E. L. Thorndike's (1898) dissertation on the comparative psychology of learning, the field seems ready for a reassessment of its metatheoretical foundations. The stability of learning phenotypes across species is shown to be similar to that of other biological characters, both genotypic (e.g., Hox genes) and phenotypic (e.g., vertebrate brain structure). Moreover, an analysis of some current lines of comparative research indicates that researchers use similar strategies when approaching problems from either an ecological view (emphasizing adaptive significance) or a general-process view (emphasizing commonality across species). An integration of learning and evolution requires the development of criteria for recognizing and studying the divergence, homology, and homoplasy of learning mechanisms, much as it is done in other branches of biological research. PMID- 11863038 TI - Enriched behavioral prediction equation and its impact on structured leaning and the dynamic calculus. AB - This theoretical note describes an expansion of the behavioral prediction equation, in line with the greater complexity encountered in models of structured learning theory (R. B. Cattell, 1996a). This presents learning theory with a vector substitute for the simpler scalar quantities by which traditional Pavlovian-Skinnerian models have hitherto been represented. Structured learning can be demonstrated by vector changes across a range of intrapersonal psychological variables (ability, personality, motivation, and state constructs). Its use with motivational dynamic trait measures (R. B. Cattell, 1985) should reveal new theoretical possibilities for scientifically monitoring change processes (dynamic calculus model: R. B. Cattell, 1996b), such as encountered within psychotherapeutic settings (R. B. Cattell, 1987). The enhanced behavioral prediction equation suggests that static conceptualizations of personality structure such as the Big Five model are less than optimal. PMID- 11863039 TI - Genes, interactions, and the development of behavior. AB - Explaining how genes influence behavior is important to many branches of psychology, including development, behavior genetics, and evolutionary psychology. Presented here is a developmental model linking the immediate consequence of gene activity (transcription of messenger RNA molecules from DNA sequences) to behavior through multiple molecular, cellular, and physiological levels. The model provides a level of detail appropriate to theories of behavioral development that recognizes the molecular level of gene action, dispensing with the metaphorical use of such terms as blueprints, plans, or constraints that has obscured much previous discussion. Special attention is paid to the possible role of immediate-early genes in initiating developmental responses to experience, adding specificity to the claim that neither genes nor experience act alone to shape development. PMID- 11863040 TI - A unified theory of implicit attitudes, stereotypes, self-esteem, and self concept. AB - This theoretical integration of social psychology's main cognitive and affective constructs was shaped by 3 influences: (a) recent widespread interest in automatic and implicit cognition, (b) development of the Implicit Association Test (IAT; A. G. Greenwald, D. E. McGhee, & J. L. K. Schwartz. 1998), and (c) social psychology's consistency theories of the 1950s, especially F. Heider's (1958) balance theory. The balanced identity design is introduced as a method to test correlational predictions of the theory. Data obtained with this method revealed that predicted consistency patterns were strongly apparent in the data for implicit (IAT) measures but not in those for parallel explicit (self-report) measures. Two additional not-yet-tested predictions of the theory are described. PMID- 11863041 TI - Reassessing working memory: comment on Just and Carpenter (1992) and Waters and Caplan (1996). AB - M. A. Just and P. A. Carpenter's (1992) capacity theory of comprehension posits a linguistic working memory functionally separated from the representation of linguistic knowledge. G. S. Waters and D. Caplan's (1996) critique of this approach retained the notion of a separate working memory. In this article, the authors present an alternative account motivated by a connectionist approach to language comprehension. In their view, processing capacity emerges from network architecture and experience and is not a primitive that can vary independently. Individual differences in comprehension do not stem from variations in a separate working memory capacity; instead they emerge from an interaction of biological factors and language experience. This alternative is argued to provide a superior account of comprehension results previously attributed to a separate working memory capacity. PMID- 11863042 TI - Models of ecological rationality: the recognition heuristic. AB - One view of heuristics is that they are imperfect versions of optimal statistical procedures considered too complicated for ordinary minds to carry out. In contrast, the authors consider heuristics to be adaptive strategies that evolved in tandem with fundamental psychological mechanisms. The recognition heuristic, arguably the most frugal of all heuristics, makes inferences from patterns of missing knowledge. This heuristic exploits a fundamental adaptation of many organisms: the vast, sensitive, and reliable capacity for recognition. The authors specify the conditions under which the recognition heuristic is successful and when it leads to the counterintuitive less-is-more effect in which less knowledge is better than more for making accurate inferences. PMID- 11863043 TI - A contour propagation approach to surface filling-in and volume formation. AB - A new approach to surface and volume formation is introduced in response to the question, "Why do some silhouettes look 3 dimensional (3D) and others look 2D?" The central idea is that form information can propagate away from a "propagable segment" (PS) of occluding contour that could have projected onto the image from the visible portion of a cross-section of a surface. A key property of a PS is that it exhibits abrupt curvature changes where it meets the rest of the occluding contour. An algorithm is described for filling in curved surfaces from a PS: When copies of a PS are propagated into the interior, they act as cross sectional surface contours that also exhibit abrupt curvature changes with the rest of the occluding contour. The result is a nonmetric coding of 3D-shape in terms of local ordinal surface curvature and orientation relationships that is scale, translation, and rotation invariant. PMID- 11863045 TI - Concluding remarks: value-driven health care--a unifying concept. PMID- 11863044 TI - About the impossibility of a single (ex-)user and survivor of psychiatry position. AB - OBJECTIVE: To present the European Network of (ex-)users and survivors of psychiatry and discuss issues of concern among (ex-)users and survivors. METHOD: Material from papers and documents published by and discussed among (ex-)users and survivors of psychiatry is used to outline topics of interest. RESULTS: The European Network of (ex-)Users and Survivors of Psychiatry (ENUSP) was founded in 1991 as an association of national/regional organizations of (ex-)users and survivors of psychiatry. The network organizes biennial conferences, all delegates are (ex-)users and survivors of psychiatry. ENUSP aims include the fight against discrimination of people with experience of the psychiatric system, support for (ex-)user/ survivor organizations, influence on policy-making, legislation and human rights debates, demedicalizing psychiatry, and opposition to unidimensional approaches to mental and emotional distress. CONCLUSION: (Ex )users and survivors should be involved in monitoring psychiatric services, education and examination of mental health professionals. User/survivor controlled services constitute an important innovative service. PMID- 11863046 TI - Mental health in Europe: problems, advances and challenges. AB - OBJECTIVE: To describe mental health care needs and challenges across the WHO European region of 51 nations. METHOD: Based on morbidity and mortality data from HFA Statistical Database and Health2l, the policy framework of WHO Europe, major trends in mental health care needs, psychiatric reform and mental health promotion are discussed. RESULTS: There is a mortality crisis related to mental ill health in Eastern European populations of transition. Destigmatization is required to improve early intervention and humanization of services, and national mental health audits are needed to create the basis for national mental health planning, implementation and monitoring. There are both problems and advances in service restructuring, and comprehensive mental health promotion programmes, preventive and monitoring strategies are required. CONCLUSION: Partnerships between national and international organizations, especially WHO and the European Union, have to be strengthened to make progress on the way to integrated community mental health services. PMID- 11863047 TI - Mental health reforms in Eastern Europe. AB - OBJECTIVE: To describe the background in general culture, public and professional discourse against which mental health care reform initiatives in Eastern Europe need to be seen. METHOD: An account of some key aspects of sociopolitical and cultural transition in Eastern European countries is given, and core results of a research project on attitudes and needs assessment in psychiatry in six Eastern European countries are reported. RESULTS: In post-totalitarian cultures mental health reforms impinge on imagination in ways which are not easy to predict. Some of the reasons for this are traced to the psychiatric practices under the system of total control, e.g. dispensary care, political abuse, reification of classificatory terms. Data on a study of attitudes suggest that institutions had replaced community life in those parts of Europe. CONCLUSION: It is predicted that with time trust in the capacity of community to contain mental illness will be regained. PMID- 11863048 TI - Psychiatric reform in Germany. AB - OBJECTIVE: To describe the ongoing process of German psychiatric reform and the structure and functioning of mental health services. METHOD: Information sources used include official reports describing mental health services, relevant publications related to organization and functioning of services. RESULTS: There has been far-reaching change in mental health care since the late 1960s: psychiatric hospitals have lost about 50% of their beds and one psychiatric hospital has been closed. One hundred and sixty-five general hospital psychiatric units have been built up. Out-patient, community and residential services have been developed. There is a lack of diversified residential and rehabilitation services, particularly for the most severely ill. Co-ordination of care is not always ensured, transfer of patients to remote nursing homes has occurred in some places. Carers and service users articulate their views to an increasing degree. CONCLUSION: Political and professional enthusiasm have been important in implementation of the German reform. Evolving it further will require major efforts. PMID- 11863049 TI - Mental health care in Germany: carers' perspectives. AB - OBJECTIVE: To assess the mental health care system in Germany from the point of view of the federal association of family carers of people with mental illness. METHOD: Family carer involvement and perspective are discussed on the basis of available literature, questionnaire surveys and documents of carer organizations. RESULTS: At the beginning of the reform movement the views of informal carers were not discussed. Since 1985 family carers have joined forces to express their views on needs of the severely mentally ill and their carers. Their aim is to point out deficits of the care system and to work towards improved care for their relatives with mental illness and changes in the mental health care system. CONCLUSION: In the reform process informal carers should receive support and be respected as experts and partners. PMID- 11863050 TI - Italian psychiatric reform 20 plus years after. AB - OBJECTIVE: To describe the current situation of mental health care in Italy and implementation of mental health reform legislation. METHOD: The current mental health care system and studies of the implementation of psychiatric reform are described. RESULTS: The 1978 reform law inaugurated fundamental changes in the care system (prohibiting admissions to state mental hospitals, stipulating community-based services, allowing hospitalization only in small general-hospital units). Uneven reform implementation was reported initially. However, in 1984 in- and out-patient services in the community were available to >80% of the population. There is a comprehensive network of in- and out-patient, residential and semi-residential facilities. Recently, services have been jeopardized by the managed-care revolution, and non-profit organizations supplement the public system (especially residential care, employment and self/mutual help). CONCLUSION: Implementation of the psychiatric reform law has been accomplished, and the year 1998 marked the very end of the state mental hospital system in Italy. PMID- 11863051 TI - Mental health service provision in England. AB - OBJECTIVE: To describe mental health service provision for adults of working age in England. METHOD: Services in an inner London area are described so as to illustrate current patterns of service organization in England. National trends are then discussed. RESULTS: Despite relatively low public expenditure, substantial progress has been made in deinstitutionalization and development of comprehensive community-based services. Persisting difficulties include high staff turnover, a minority of patients. who do not engage with community services, user and carer dissatisfaction with emergency services, and social exclusion because of stigma. Recent government policy advocates resolving some of these problems using new service models such as assertive outreach and crisis teams. CONCLUSION: Closure of the large asylums has largely been accomplished. England is now entering a new phase in community service development, with a range of innovative developments aimed at resolving problems still encountered after the initial phases of integrated community service development. PMID- 11863052 TI - Psychiatric reform in Russia. AB - OBJECTIVE: To describe the mental health care system in Russia against the background of rapid social, economic and political change since the late 1980s. METHOD: Indicators of social stress and deteriorating population mental health and official data on service provision are presented. RESULTS: In the current system of psychiatric care dispensaries take a central position. Key issues of the discussion on mental health care include ways of coping with social stress disorders, strategies to redefine psychiatric rehabilitation in a changed social economic context and steps towards strengthening social support networks for people with mental illness. Following political abuse of psychiatry, professionals have had to face justified and unjustified accusations, and been faced with a general challenge to their role. CONCLUSION: Tackling problems of social stress, the integration of mental health care in the general medical care system and the building-up of general hospital in-patient psychiatric units are of strategic importance. PMID- 11863053 TI - Trends and developments in public psychiatry in France since 1975. AB - OBJECTIVE: The authors present an account of the current state of sectorization in France and its development over the last decade. METHOD: The paper is based on statistics collected by the French Health Ministry and on relevant laws and regulations. Comments by patients, families and professionals are presented. RESULTS: The supply of mental health care is usually satisfactory, and there is a tendency for in-patient units to be converted into day hospitals. There is a large disparity between regions, especially in child and adolescent psychiatry. The 1838 law governing compulsory admissions was revised in 1990, and this revision was the object of a commission of inquiry in 1995. Psychiatric units work on the borders between health and social services and are forging new links between the two. CONCLUSION: Psychiatric care requires an integrated health and social perspective. In the field of mental health legislation European recommendations are called for. PMID- 11863054 TI - Evolution of the mental health care system in Poland. AB - OBJECTIVE: The aim of this paper is to trace evolution of mental health system in Poland. METHOD: Available Polish literature and fundamental policy documents including mental health legislation are reviewed and major milestones in this evolution identified and then discussed against the background of political and social developments. RESULTS: The mental health system evolved since the beginning of the 1970s from large hospitals towards community-based care. It was found that the changes were rather slow due to financial shortages and lack of clear demand from users. Recent transitions offer opportunities to increase impact of users and their families. On the other hand, however, introduction of health insurance system reduces prospects for well co-ordinated mental health policy and may lead to inequalities in access to services. CONCLUSION: It can be concluded that evolution of mental health services has been influenced by prevailing ideologies as much as by technical and professional considerations. PMID- 11863055 TI - Mental health care reform in The Netherlands. AB - OBJECTIVE: To describe the major changes in mental health care for adults in the Netherlands during the past 25 years. METHOD: Scientific literature and official documents. RESULTS: Phases of the reform process are the integration of ambulatory services in the early 1980s and the following implementation of community mental health centres (RIAGGs); the differentiation and extramuralization of mental hospitals; the differentiation within the field of living accommodations; and the final fusion process between these three into integrated regional mental health care organizations. Current issues in the development of services are, e.g. the ever growing demand for mental health care, special programmes for defined target populations, legislation and patient rights, rehabilitation and empowerment. CONCLUSION: The Dutch mental health care system has a low threshold and a comparatively good quality. There is a long lasting and strong influence of user and family organizations on the content and quality of services. Recently important organizational changes are taking place. PMID- 11863056 TI - The European perspective of psychiatric reform. AB - OBJECTIVE: To provide a framework of mental health care reform across Europe. METHOD: On the basis of summary quantitative indices and expert ratings of broad aspects of mental health care structure, the process and outcome of psychiatric reform common trends and differences are outlined. RESULTS: There has been a broad trend away from an institutional model of care with the mental hospital as the dominant institution, and community- and general hospital-based mental health services of varying comprehensiveness are in place in most countries. The social and broad community aspects of psychiatric reform have generally been somewhat less successful than changes in service set-up. Assessment of reform outcomes proves particularly difficult. CONCLUSION: Psychiatric reform processes have achieved some of their aims, and there are broadly similar trends. Regional variation is substantial and may be as important as cross-national differences. Mental health care reform is ongoing across the European region. PMID- 11863057 TI - Mental health care reform in Sweden, 1995. AB - OBJECTIVE: To describe the content of the Community Mental Health Care reform in Sweden, in effect from 1995 and directed to severely mentally ill people (SMI). METHOD: Evaluating changes, at local and national level, in living conditions among SMI and resources of services directed to them, by using registers, questionnaires, interviews and case studies. RESULTS: A survey, covering 93% of the population, identified 43,000 SMI (prevalence of 0.63%); 4,000 long-stay patients and 400 rehabilitation programmes were transferred from psychiatric services to social services (15% of the budget of psychiatric services). Employment and rehabilitation projects, family support and user programmes and educational projects for social services staff, were launched (funded by state subsidies). CONCLUSION: SMI still have difficulties in obtaining adequate support on the basis of disability laws and there continue to be barriers between social services and psychiatric services. PMID- 11863058 TI - Spanish psychiatric reform: what can be learned from two decades of experience? AB - OBJECTIVE: The objective of the paper is to describe the impact of Spanish psychiatric reform on the organization and functioning of mental health services. METHOD: This paper is based on official administrative reports and on relevant related publications. RESULTS: The most significant achievements of Spanish psychiatric reform have been: (i) the development of a new organization of mental health care, decentralized in character and territorially based; (ii) the integration of psychiatric patients in general health care; (iii) the creation of an extensive community network of health centres; and (iv) the development of more positive attitudes towards mental illness. However, our analysis also reveals the existence of significant deficiencies. CONCLUSION: Analysis of the Spanish experience shows that the process of psychiatric reform depends basically on long-term commitments, which in a system such as Spain's must come from central administration and also from the autonomous communities. PMID- 11863059 TI - European perspectives: a carer's view. AB - OBJECTIVE: To present the work of the European Federation of Associations of Families of Mentally III People (EUFAMI) and discuss issues of concern to family carers. METHOD: The problem areas identified and discussed by family carers are presented on the basis of questionnaire surveys organized by EUFAMI. Addresses of national organisations of family carers are included. RESULTS: A range of problem areas are identified; they include subsistence and welfare payments for the severely mentally ill, some shortage of general hospital units, problems of care co-ordination, issues of respect for family carers and family involvement. CONCLUSION: The aim of best practice in mental health care throughout Europe has not yet been reached. Key activities of EUFAMI are aimed at empowerment of families and best practice in psychiatry in Europe. PMID- 11863060 TI - Exploring racial variations in the spousal sex ratio of killing. AB - The following article examines differences in the social situation of intimate partners as an explanation of racial differences in the female to male ratio of spousal homicides in Canada. An analysis of homicide data from 1961 to 1983 generated by the Canadian Centre for Justice Statistics reveals that the ratio of women killing their husbands to men killing their wives is highest for Aboriginals and lowest for Blacks, with the ratio for Whites falling in between. The possible sources of racial differences in this ratio include the proportion of couples (a) in common-law relationships, (b) who are co-residing as opposed to being separated, and (c) for whom there is a substantial age disparity between the partners. These factors are related to the spousal sex ratio of killing more generally. An exploration of interracial homicide patterns and racial variation in jealousy-motivated homicides was also undertaken. The findings reveal that controlling for the above factors substantially reduces the importance of race in predicting the gender of the homicide victim. PMID- 11863061 TI - The influence of victim gender and sexual orientation on judgments of the victim in a depicted stranger rape. AB - This study investigated the impact of respondent gender, victim gender, and victim sexual orientation on judgments toward the victim of a depicted stranger rape. Respondents were required to read a scenario in which victim gender and sexual orientation varied between subjects, and to complete measures of behavioral blame, responsibility, and severity of the attack. Results revealed that male respondents made more anti-victim judgments than female respondents did. Male respondents judged gay male victims more negatively than they did other victims. Female respondents' judgments were pro-victim regardless of victim gender and victim sexual orientation. Results are discussed in relation to the feminist analysis of victim blame, and blame toward male rape victims. Implications for support services, particularly of male victims, are also considered. PMID- 11863062 TI - Justifiability, sympathy level, and internal/external locus of the reasons battered women remain in abusive relationships. AB - Reasons why battered women remain in abusive relationships were rated by college students as to the degree of sympathy the reasons engendered, the degree to which the reasons seemed to justify the woman remaining, and whether the reasons appeared more within the woman's control (internal) or imposed upon her (external). Four clusters of reasons emerged that varied on the sympathy and justifiability ratings: Physical Restraint/Lack of Resources; Maintenance of the Relationship; Personal Deficits; and Loss of Resources. Sympathy for the individual reasons was correlated with viewing these reasons as more justifiable for keeping her in the relationship. Both sympathy and justifiability ratings were higher if the reasons were seen as externally imposed on the woman. The more the participants viewed the battered woman as possessing socially desirable traits, the more they had sympathy for the reasons she stayed and viewed them as justifiable. Gender and ethnicity differences on the ratings were also found. Implications for future research and application of the findings are discussed. PMID- 11863063 TI - A follow-up study of posttraumatic stress disorder, anxiety, and depression in Australian victims of domestic violence. AB - Although posttraumatic stress disorder (PTSD), anxiety, and depression are acknowledged consequences of domestic violence, little information is available on the course of recovery over time and factors that may mediate positive outcome. Fifty-nine women were assessed for the presence of PTSD and levels of anxiety and depression at time of shelter residence and again one year later. Results at follow-up indicated a significant reduction in the incidence of PTSD, although a substantial number of women continued to report a range of posttrauma symptoms. There were also significant reductions in the levels of anxiety and depression over the 12-month period. Findings indicated the particular importance of safety and the presence of social support as prerequisites for recovery. PMID- 11863064 TI - Sexual abuse: a comparison between resilient victims and drug-addicted victims. AB - The goal of this study was to determine which variables distinguish resilient victims from drug-addicted victims, who were sexually abused during their childhood--in addition, to measure the contribution of these variables to the level of distress experienced by the victims. There were two groups of 20 women interviewed. The resilient group showed no clinically significant symptoms of mental distress, and the addicted group were undergoing treatment for drug dependency. They all completed a semi-structured interview and a questionnaire regarding the type and severity of their sexual abuse, mental health status, self esteem, locus of control, support and cognitive factors from Finkelhor's model. Both of these groups were equally and severely abused. Resilient and addicted women both received a moderate level of support. These women also reported the same sense of betrayal and powerlessness. Furthermore, both groups believe, to a large degree, that they now control what happens to them (internal locus of control). There were three distinguishing variables among the two groups, they were stigmatization, self-blame, and hazard for the locus of control. In comparison, resilient women had less self-blame for having been abused and they also felt less stigmatized than addicted women. In fact, stigmatization and self blame account for 65% of the TSC-40 variance. These results suggest that cognitive strategies, particularly those that are linked to the interpretation of the event, may have some importance in the recovery. PMID- 11863065 TI - Social reactions to sexual assault victims from various support sources. AB - This study examined positive and negative social reactions received by 323 victims disclosing sexual assaults to informal and formal support providers. Analyses of both quantitative and qualitative data indicated that both positive and negative reactions are commonly received by victims disclosing their assaults to others, but that victims seeking help from formal support sources are more commonly faced with negative reactions of victim blame, stigmatizing responses, and controlling reactions from others. Qualitative analysis of written responses provided by respondents also revealed other less common but unhelpful reactions of assault disclosure such as rape myths, violations of the victim's confidential disclosures, and revictimization. Preliminary analyses of the effects of specific reactions on adjustment from different types of support providers suggested that reactions of friends are particularly important for recovery. Positive reactions from friends appear to be more helpful, whereas negative reactions appear to be harmful from both informal and formal support sources. Implications for research and treatment of sexual assault victims are discussed. PMID- 11863066 TI - The program effect of batterer programs in three cities. AB - Recent meta-analyses and experimental designs of batterer program evaluations suggest little or no program effect. This finding may be compromised by a variety of analytical issues. Instrumental variable analysis addresses some of these issues, especially the relationship of program dropout to batterer reassault. This method of analysis was, therefore, used to test for program effect in a multi-site evaluation. The sites were three well-established batterer programs using a gender-based, cognitive-behavioral approach (n = 640). Completing a batterer program reduced the likelihood of reassault by 44% to 64%, depending on the specification used. Completing a 3-month program appeared to be as effective as completing a 5 1/2- or 9-month program. This moderate effect is an accomplishment considering the problems associated with the program participants and the inconsistency in the criminal justice system at the research sites. The findings remain tentative because of weak instrumental variables for reassault, but do confirm the need for more complex analyses of program effect. PMID- 11863068 TI - Fragile sites in human and Macaca fascicularis chromosomes are breakpoints in chromosome evolution. AB - We have analysed the expression of aphidicolin-induced common fragile sites at two different aphidicolin concentrations (0.1 micromol/L and 0.2 micromol/L) in three female and one male crab-eating macaques (Macaca fascicularis, Cercopithecidae, Catarrhini). A total of 3948 metaphases were analysed: 1754 in cultures exposed to 0.1 micromol/L aphidicolin, 1261 in cultures exposed to 0.2 micromol/L aphidicolin and 933 in controls. The number of breaks and gaps detected ranged from 439 in cultures exposed to 0.1 micromol/L aphidicolin to 2061 in cultures exposed to 0.2 micromol/L aphidicolin. The use of a multinomial FSM statistical model allowed us to identify 95 fragile sites in the chromosomes of M. fascicularis, of which only 16 are expressed in all four specimens. A comparative study between the chromosomes of M. fascicularis and man has demonstrated that 38 human common fragile sites (50%) are found in the equivalent location in M. fascicularis. The analysis of the rearrangements that have taken place during chromosome evolution has revealed that the breakpoints involved in these rearrangements correspond significantly (p < 0.025) to the location of M. fascicularis fragile sites. PMID- 11863067 TI - Chromosome ends in Chironomus tentans do not have long single-stranded overhangs characterizing canonical telomeres. AB - Single-stranded overhangs of the G-rich strand belong to the conserved features of telomeres composed of short telomeric repeats. These structures are thought to be essential for the maintenance of proper telomeric structure and function and the mechanism of their generation is telomerase-independent. We have examined the presence of single-stranded overhangs in Chironomus tentans, a dipteran insect lacking canonical telomeres that uses 350-bp repeats to terminate its chromosomes. Using a non-denaturing in-gel hybridization technique, we found that C. tentans telomeres are unlikely to have single-stranded overhangs longer than 30 nt found in most other higher eukaryotes. These differences might reflect special capping mechanisms for telomeres terminated with long complex repeats. PMID- 11863069 TI - Meiotic chromosomes as templates for microdissection. AB - As templates for chromosome microdissection, meiotic cells offer several advantages over mitotic cells. The pairing of homologous chromosomes at the metaphase plate of the first meiotic division allows the simultaneous isolation of two copies of the same chromosome, and the sex chromosomes are easy to identify in male meiotic cells. We report on a method for making fluorescence in situ hybridization (FISH) probes from dissected meiotic chromosomes. PMID- 11863070 TI - In-situ comparative mapping (ISCM) of Glu-1 loci in Triticum and Hordeum. AB - The ability to detect small low- or single-copy DNA sequences by fluorescence in situ hybridization (FISH) is an important step towards physical mapping of plant genomes. In this study, the FISH technique was used to physically map the Glu-1 loci controlling high-molecular weight (HMW) glutenin in common wheat (Triticum aestivum cv. 'Chinese Spring') and tritordeum (an amphiploid between T. turgidum cv. durum and Hordeum chilense). The probe used was the single-copy Glu-D1-1d gene coding the 1Dx5 HMW glutenin subunit. Three loci were mapped on chromosomes of wheat homoeologous group 1 (arm 1AL, 1BL and 1DL). The Glu-1 loci were mapped (fraction of the distance from the centromere) at positions 0.76 +/- 0.01, 0.69 +/- 0.01 and 0.76 +/- 0.01, on arms 1AL, 1BL and 1DL, respectively. The Glu-1 loci were also mapped on chromosomes of homoeologous group 1 of tritordeum at positions 0.75 +/- 0.01, 0.70 +/- 0.01 and 0.60 +/- 0.01, on arms 1AL, 1BL and 1HchL, respectively. Chromosomes with positive signals were identified by reprobing chromosome preparations using both the GAA-satellite and pAs1 sequences simultaneously. The application of the FISH technique to study homoeology among different genomes is discussed. PMID- 11863071 TI - Principles of the highly ordered arrangement of metaphase I bivalents in spermatocytes of Agrodiaetus (Insecta, Lepidoptera). AB - We have investigated the nature of highly ordered bivalent arrangement in lepidopteran spermatocytes by analysing and comparing the patterns of bivalent distribution in intact metaphase I plates of 24 closely related species of the genus Agrodiaetus (Lycaenidae). The studied species greatly differed in haploid chromosome numbers (from n = 13 to n = 90) and in the structure of their karyotypes. We found that the larger the bivalent, the closer to the centre of the metaphase plate it was situated. In species with a high chromosome number and asymmetrical karyotype structure, the largest bivalent was located in the centre of the circular metaphase plate. Bivalents of equal size were approximately equidistant from the centre of the metaphase plate and formed concentric circles around the largest bivalent. These principles are diametrically different from those known in the majority of other animals and plants, in which the smallest elements of the chromosome set are situated in the centre of metaphase plate. The only exception from the above principles was observed in spermatocytes of A. surakovi which were heterozygous for reciprocal translocation involving two or three chromosome pairs. In addition to one large bivalent, the heterozygous cells had a multivalent, the size of which was comparable to or even exceeded that of the largest bivalentin the karyotype. In spite of thelarge size, the multivalent was always situated at the periphery of metaphase plate. This indicated that the chromosome size itself is not the only factor determining the bivalent position. We also found that the structure of the metaphase plate is fundamentally different in mitotic and meiotic cells of Agrodiaetus. In spermatogonial metaphase, chromosomes were tightly brought together, forming a dense compact disk, whereas during metaphase I of spermatocytes, all bivalents were clearly separated from each other, and the distance between adjacent bivalents varied from 0.4 to 1.5 microm. Based on the above findings, we proposed a model of bivalent distribution in the Lepidoptera. According to the model, during congregation in the prometaphase stage there is a centripetal movement of bivalents made by a force directed to the centre of the metaphase plate transverse to the spindle. This force is proportional to the kinetochore size of a particular bivalent. The Lepidoptera have a special near-holokinetic type of chromosome organisation. Therefore, large bivalents having large kinetochores are situated in the central part of metaphase plate. Another possible factor affecting the bivalent position is the interaction of bivalents with the cisternae of the membrane system compartmentalising the intraspindle space. PMID- 11863072 TI - Direct evidence for the Homo-Pan clade. AB - For a long time, the evolutionary relationship between human and African apes, the 'trichotomy problem', has been debated with strong differences in opinion and interpretation. Statistical analyses of different molecular DNA data sets have been carried out and have primarily supported a Homo-Pan clade. An alternative way to address this question is by the comparison of evolutionarily relevant chromosomal breakpoints. Here, we made use of a P1-derived artificial chromosome (PAC)/bacterial artificial chromosome (BAC) contig spanning approximately 2.8 Mb on the long arm of the human Y chromosome, to comparatively map individual PAC clones to chromosomes from great apes, gibbons, and two species of Old World monkeys by fluorescence in-situ hybridization. During our search for evolutionary breakpoints on the Y chromosome, it transpired that a transposition of an approximately 100-kb DNA fragment from chromosome 1 onto the Y chromosome must have occurred in a common ancestor of human, chimpanzee and bonobo. Only the Y chromosomes of these three species contain the chromosome-1-derived fragment; it could not be detected on the Y chromosomes of gorillas or the other primates examined. Thus, this shared derived (synapomorphic) trait provides clear evidence for a Homo-Pan clade independent of DNA sequence analysis. PMID- 11863073 TI - Chromosome sorting and PCR-based physical mapping in pea (Pisum sativum L.). AB - Three pea lines with reconstructed karyotypes were used for analysis and subsequent purification of individual chromosome types using flow cytometry and sorting. The lines JI 145, JI 146, and JI 148 possess defined chromosomal translocations allowing discrimination of three to four chromosome types from each line based on the different sizes of translocation chromosomes. Whereas only two chromosomes could be sorted from standard (wild-type) karyotype, a combined use of these lines allowed sorting of six out of the seven types of pea chromosomes. Chromosomes were identified and purity of flow-sorted fractions was assessed using fluorescence in-situ hybridization with a PisTR-B probe that was previously shown to give labelling patterns characteristic for each chromosome type. The fractions of flow-sorted chromosomes were of very high purity (> 95%) and proved to be suitable for detection of gene and marker sequences using PCR with specific primers. Three fractions containing chromosomes 27, 72 and a pool of all remaining chromosomes (1, 3, 4, 5, 6) flow-sorted from the line JI 148 were then used for PCR-based physical localization of genetic markers selected from linkage groups IV and VII. These experiments enabled assignment of the linkage groups IV and VII to chromosomes 4 and 7, respectively. PMID- 11863075 TI - The nature and destiny of translocated B-chromosome-specific satellite DNA of rye. AB - Translocations of A chromosome-specific and B chromosome-specific satellite DNA were tracked by fluorescence in situ hybridisation from an irradiated M1 generation of an experimental population of rye (Secale cereale L.) to its M2 progeny. Although high frequencies of large structural rearrangements were detected in root-tip meristems of M1 plants, none was present at meiosis or in somatic cells of their progeny. These results are interpreted in terms of efficient "filtering" of translocations during vegetative development, and not in the more usual terms of meiosis presenting a physical barrier to structural variants. These observations highlight the fact that B-A translocations are not tolerated, and may explain why this form of chromosome mutation is largely absent from natural populations. PMID- 11863074 TI - Partially inverted tandem repeat isolated from pericentric region of chicken chromosome 8. AB - The majority of chicken repetitive sequence is nuclear-membrane-associated sequence (CNM), which resides in a large number of microchromosomes (chromosomes 11-39) and is absent from macrochromosomes 1-5, ZW, and some of the intermediate chromosomes 6-10. Two repetitive families, EcoRI/XhoI, are confined to the female specific W chromosome. The core repeat units of the three families are 21 bp, containing (A)3-5 and (T)3-5 clusters separated by 5-7-bp sequences. In this article, we describe the isolation and initial characterization of a novel repeat family that is related to CNM/EcoRI/XhoI families. The novel family, designated as PIR, consists of multiple types of partially inverted repeat units of about 1.2, 1.4 and 1.6 kb. The PIR sequence is restricted to chicken chromosome 8, and accounts for about 3.8 mb, or 2500 copies of the 1.4-kb units, of the chicken genome. The evolution of PIR and related sequences is discussed. PMID- 11863076 TI - End-stage renal disease survival in blacks and whites. PMID- 11863077 TI - Observations on HIV-associated renal disease in the era of highly active antiretroviral therapy. AB - BACKGROUND: Renal disease in patients infected with HIV has evolved to include several lesions, including HIV-associated nephropathy (HIV-AN), which can promote progressive loss of renal function. Although angiotensin-converting enzyme inhibitors and corticosteroids are beneficial in selected patients, the effect of highly active antiretroviral therapy (HAART) on renal function is currently being explored. METHODS: We undertook a retrospective study to determine the types of renal lesions present in patients infected with HIV with renal insufficiency and/or proteinuria during the era of HAART availability and the effect of HAART on renal outcomes in these patients. Patients with HIV infection referred to the renal clinic from July 1996 through December 2000 were evaluated. Patient characteristics and data were collected including CD4 count, viral load, serum creatinine, blood pressure, proteinuria, renal ultrasound, and biopsy results, and treatment with HAART. Study endpoints were doubling of serum creatinine, initiation of dialysis, or death. RESULTS: Twenty-three patients met study criteria, 13 received HAART, and 10 did not. Baseline characteristics were similar except for renal function parameters, viral loads, and CD4 counts. A variety of lesions were noted on 12 renal biopsies. A clinical diagnosis of HIV AN was made in the other 11 patients. Only 2 patients receiving HAART before renal evaluation were noted to have HIV-AN. In the HAART group, none of the patients, including those with HIV-AN, developed a doubling of serum creatinine. In the non-HAART group, all patients manifested a doubling of serum creatinine, 2 patients died, and 8 patients required dialysis. CONCLUSIONS: A variety of renal lesions are noted in patients infected with HIV during the HAART era. Patients who received HAART maintained stable renal function, whereas patients who did not required dialysis therapy or died with advanced renal failure. It seems that HAART may improve renal outcomes in patients with HIV and renal disease. PMID- 11863078 TI - Wernicke encephalopathy in nonalcoholic patients. AB - BACKGROUND: Wernicke encephalopathy is associated with patients that have a history of alcohol abuse and other malnourished states. However, clinicians may be less likely to recognize this condition in nonalcoholic patients. This study was undertaken to evaluate the cause, diagnosis, treatment, and outcome of nonalcoholic patients with Wernicke encephalopathy. METHODS: A retrospective review was conducted of all cases of Wernicke disease from 1984 to 1999 evaluated at a rural university hospital. Inclusion criteria consisted of patients diagnosed clinically with this disorder who improved after the administration of thiamin. Patients were also included in the series if they had classic pathologic findings of the disease on autopsy even without a prior clinical diagnosis. RESULTS: Twelve patients were identified. Alcohol abuse was noted in only 50%. The patients without a history of alcohol abuse had other malnourished states, such as end-stage cancer, intractable vomiting after gastric reduction procedure, and prolonged hospital courses on intravenous fluids without other sources of nutrition. The mortality rate was 4 of 12 (33%) but due to multiple causes. Autopsies were performed on all 4 patients. Three of these 4 patients had no history of alcohol abuse. Two of the nonalcoholic patients were not diagnosed until autopsy, whereas all the patients with a history of alcohol abuse were diagnosed clinically. DISCUSSION: Our study reveals that Wernicke encephalopathy continues to be a rare but life-threatening condition often overlooked in the nonalcoholic population, resulting in the further progression of an easily treatable condition. PMID- 11863079 TI - Relapsing group B Streptococcal bacteremia in an adult. AB - We present a case of relapsing group B streptococcal bacteremia in a patient with cirrhosis secondary to hepatitis C. As no focal infection was found, other then cervical carriage, the patient required secondary prophylaxis with penicillin following the third episode of primary bacteremia within a twelve month time frame. PMID- 11863080 TI - Clodronate in myelofibrosis: a case report. AB - A 59-year-old man had well-documented agnogenic myeloid metaplasia (AMM) with pancytopenia. Frequent blood transfusions were required over a 10-month period. Androgen therapy was not beneficial and treatment with interferon resulted in severe thrombocytopenia with no decrease in transfusion requirements. Treatment with clodronate at a daily oral dose of 30 mg/kg resulted in a marked decrease in bone marrow fibrosis, and gradual normalization of blood counts over an 8-month period. He has been transfusion independent for the last 33 months. We support the findings of a previous case report that oral bisphosphonate therapy may be of value in patients with AMM. PMID- 11863081 TI - The epidemiology of end-stage renal disease among African Americans. AB - Although disparities in outcomes among African Americans compared with whites with respect to cardiovascular disease, cancer, diabetes, infant mortality, and other health standards have been well-described, these disparities are most dramatic with respect to kidney diseases. End-stage renal disease (ESRD) occurs almost 4 times more commonly in African Americans than in their white counterparts. These disparate rates of kidney disease may be caused by the complex interplay of genetic, environmental, cultural, and socioeconomic factors. African Americans are particularly vulnerable to the deleterious renal effects of hypertension and may require more aggressive blood pressure control than whites to accrue benefit with respect to preservation of renal function. Diabetes, the leading cause of ESRD in the United States, is another important factor in the excess renal morbidity and mortality of African Americans because of its prevalence in this population. Other renal diseases, especially those associated with HIV/AIDS, are also much more likely to affect African Americans than other American population subgroups. A more thorough understanding of the epidemiology of renal diseases in African Americans and the cultural, social, and biological differences that underlie racial disparities in prevalence of renal disease will be essential to the design of effective public health strategies for prevention and treatment of this burdensome problem. PMID- 11863082 TI - Pathophysiology of chronic progressive renal disease in the African American patient with hypertension. AB - Chronic renal failure and ESRD are major causes of morbidity, mortality, and chronic disability in patients in the United States. Hypertension is a major underlying cause of chronic progressive renal disease and continues to be a leading reason for the heavy burden of ESRD observed in African Americans. Hypertension is actually a syndrome of vascular pathology manifesting itself in patients by a constellation of common findings and attributes. These pathophysiologic alterations include dysregulation of arterial compliance, endothelial dysfunction, obesity and insulin resistance, abnormal sympathetic nervous system activation, accelerated atherosclerosis, left ventricular hypertrophy, and a propensity for increased vascular thrombogenesis among others. This review will focus on some of the important mechanisms possibly involved in the progression of renal disease in the setting of chronic hypertension. PMID- 11863083 TI - Diabetic renal disease in African Americans. AB - Diabetic nephropathy (DN) is the No. 1 cause of end-stage renal disease in the United States and is highly prevalent in African Americans. Almost all DN in African Americans is caused by type 2 diabetes. Glycemic control and control of blood pressure are essential to prolong renal survival and to protect against cardiovascular events. Among African Americans, diabetic nephropathy seems to affect women more than men, which may be related to increased rates of obesity and diabetes in African American women. In addition to gender, the development of albuminuria, family history, and possibly birth weight are factors that predict progression of renal disease in African Americans with DN. The impact of glycemic control, appropriate antihypertensives, and the optimal level of blood pressure control in African Americans with advanced DN require further study. This article will review the clinical characteristics, risk factors, predictors of disease progression, and treatment of diabetic nephropathy in African Americans. PMID- 11863084 TI - Lupus nephritis in African Americans. AB - Lupus nephritis is more common and severe in African American women. Despite aggressive immunosuppressive therapies employed in lupus nephritis, African Americans have a higher incidence of progression to end-stage renal disease. The reasons for the racial disparities are not clear, but seem to be due to genetic, environmental, and socioeconomic factors. Hypertension and proteinuria are well defined prognostic factors that significantly impact the course of renal disease progression for most forms of renal disease. However, clinical trials in lupus nephritis to date have not evaluated the role of aggressive antihypertensive or antiproteinuric therapies in retarding renal disease progression. Thus, additional studies are needed to better elucidate the natural history of lupus nephritis in African Americans and to optimize therapeutic strategies for those who are identified as being at high risk. PMID- 11863085 TI - Focal segmental glomerulosclerosis in African Americans. AB - Focal segmental glomerulosclerosis (FSGS), the leading glomerular cause of the nephrotic syndrome among African Americans, is typically associated with edema, proteinuria, hypertension, microscopic hematuria, and renal insufficiency. Recent studies suggest that either the incidence of FSGS has increased or an increased number of biopsies of African American patients have made the diagnosis more common. The collapsing variant of FSGS, which occurs more commonly in African Americans than in whites, carries an especially poor prognosis with respect to renal survival. Although the pathogenesis of FSGS is not well understood, the fact that it frequently recurs early after transplantation has led to speculation that patients with FSGS may have a circulating factor that leads to increased glomerular permeability. There are no randomized control trials of treatment regimens for FSGS. Steroids, alkylating agents, and cyclosporin have all been used with variable results to treat FSGS. PMID- 11863087 TI - Generics again... PMID- 11863086 TI - African Americans and renal transplantation: disproportionate need, limited access, and impaired outcomes. AB - BACKGROUND: Renal transplantation is the therapy of choice for patients with end stage renal disease (ESRD). However, African Americans' (AA) access to this modality is not commensurate with that of other races. This imbalance, coupled with AA disproportionately representing those with ESRD, has kept AA disadvantaged compared with other races, especially whites. METHODS: We reviewed published reports that examined the connection between race and the incidence of chronic renal failure, access to optimal therapy, and outcomes of renal transplantation. RESULTS: The incidence of ESRD in AA is 4 times greater than in whites, but AA remain less likely than whites to be referred for or undergo renal transplantation. Also, AA are at greater risk than whites to experience premature graft failure. CONCLUSIONS: ESRD management has improved dramatically with the advent of successful renal transplantation. However, AA remain significantly disadvantaged in both access and outcomes compared with whites. Further evaluation of underlying causes and development of specific remedies is warranted. PMID- 11863088 TI - Modulation of 5-fluorouracil/leucovorin by trimetrexate--did it work? PMID- 11863089 TI - Oxaliplatin plus vinorelbine in advanced non-small-cell lung cancer: final results of a multicenter phase II study. AB - BACKGROUND: Oxaliplatin and vinorelbine are both active agents against non-small cell lung cancer (NSCLC). In a previous phase I trial, we showed that oxaliplatin (130 mg/m2, day 1) and vinorelbine (26 mg/m2/day, days 1 and 8) can be safely combined when given every 21 days. We completed the evaluation of this new platinum-based doublet in advanced NSCLC patients in a multicenter phase II study. PATIENTS AND METHODS: Twenty-eight chemotherapy-naive patients (22 men and six women: median age 58 years, range 33-70), including 20 with stage IV disease, received this out-patient combination, with 5-hydroxytryptamine-3-receptor agonists as the only prophylactic measure. RESULTS: A total of 117 cycles were given, for a median of three per patient (range 1-8). Of 26 eligible patients, nine achieved a partial response (WHO criteria), giving an objective response rate of 35% [95% confidence interval (CI) 17% to 56%]. The median progression free survival was 5.0 months (95% CI 3.1 to 6.9), median overall survival was 9.8 months (95% CI 2.2 to 17.5) and the 1-year survival rate was 37%. Neutropenia was the principal toxicity, grade 4 occurring in 11 patients (39%) and 25 cycles (22%). Four patients (14%) experienced one episode of febrile neutropenia each. Acute oxaliplatin-related neurosensory toxicity was prevalent, but was mild to moderate in the majority of patients (82%) and reversible. Grade 1/2 vomiting (65% of patients) and diarrhea (32% of patients) were easily managed. CONCLUSIONS: The oxaliplatin-vinorelbine doublet is a safe and active out-patient combination. It may represent an interesting alternative in the management of patients with NSCLC, and serve as a new doublet to which other active agents could be added. PMID- 11863090 TI - Paclitaxel plus carboplatin, compared with paclitaxel plus gemcitabine, shows similar efficacy while more cost-effective: a randomized phase II study of combination chemotherapy against inoperable non-small-cell lung cancer previously untreated. AB - BACKGROUND: Paclitaxel (Taxol) plus carboplatin (PC) has shown activity in the treatment of advanced non-small-cell lung cancer (NSCLC). Non-platinum-containing combination chemotherapy, such as paclitaxel plus gemcitabine (PG), has also demonstrated reasonable efficacy. Our aim here was to evaluate the clinical efficacy and cost-effectiveness of PC versus PG in chemo-naive. advanced NSCLC patients. PATIENTS AND METHODS: Ninety (68 male, 22 female) patients were enrolled from August 1999 to August 2000. The performance status was one in 29 patients and two in 16 patients of the PC group, and one in 24 patients and two in 21 patients of the PG group. Seventeen patients had stage IIIb disease and 28 patients stage IV disease in the PC group: 18 patients had stage IIIb disease and 27 patients stage IV disease in the PG group (New International Staging System). Treatment consisted of P 175 mg/m2 and C at AUC = 7 (predicted using measured clearances and the Calvert formula) intravenous infusion (i.v.) on day 1, or P 175 mg/m2 i.v. on day 1 and G 1000 mg/m2 i.v. on days 1 and 8, every 3 weeks. RESULTS: In all, 175 cycles of PC and 184 cycles of PG were given in the PC and PG groups, respectively. The median treatment cycle was four cycles in both groups. All the patients were assessable for toxicity and response measurement. There were three complete responses and 15 partial responses (overall 40%) in the PC group, and no complete response, but 18 partial responses (overall 40%) in the PG group. WHO grades 3/4 leukopenia, anemia and thrombocytopenia occurred in six (13.3%), seven (15.5%) and five patients (11.1%) in the PC group; and in four (8.9%), six (13.3%) and 0 patients in the PG group, respectively. Two patients in each group suffered from grade 3 peripheral neuropathy. Other non-hematological toxicities were mild and few. Median survival time was 14.1 months in the PC group and 12.6 months in the PG group. One-year survival was 50.7% in the PC group and 53.3% in the PG group. The PG group had a higher total expense and expended more days undergoing treatment than the PC group (P = 0.034 and 0.069, respectively). CONCLUSIONS: Both PC and PG combination chemotherapy produce a similar efficacy in the treatment of NSCLC. However, PC is more cost-effective than PG. PMID- 11863091 TI - A phase II trial of weekly intravenous gemcitabine and cisplatin with continuous infusion fluorouracil in patients with metastatic renal cell carcinoma. AB - BACKGROUND: We reported previously that the combination of gemcitabine and continuous infusion fluorouracil (5-FU) has activity in renal cell carcinoma. Based upon in vitro synergy of gemcitabine/cisplatin and 5-FU/cisplatin, we hypothesized that the addition of cisplatin could improve the objective response rate of gemcitabine and 5-FU with manageable toxicity. PATIENTS AND METHODS: Twenty-one patients with metastatic renal cell carcinoma (RCC) and a Cancer and Leukemia Group B performance status of 0 to 2 were enrolled. Ten had received prior systemic therapy. Treatment consisted of gemcitabine 600 mg/m2 and cisplatin 20 mg/m2 on days 1, 8 and 15 of each 28-day cycle. Continuous infusion 5-FU was given from day 1 to day 21. RESULTS: No complete responses and one partial response were observed for an objective response rate of 5% (95% confidence interval 0% to 24%). Two minor responses (25% to 50% regression) were also observed. The median overall survival was 10 months with 35% of patients surviving at 1 year. Grade 3-4 myelosuppression (mostly thrombocytopenia) occurred in nine (43%) patients. Nausea/vomiting and neuropathy were dose limiting in an additional five patients. Only 51% of treatment cycles were delivered on time and without dose reduction. CONCLUSIONS: The addition of cisplatin to gemcitabine and 5-FU did not improve the objective response rate of gemcitabine and 5-FU alone and added to the toxicity. Due to the cumulative toxicity, further trials with this cisplatin-containing regimen in RCC are not indicated. PMID- 11863092 TI - Continuous low-dose anti-angiogenic/ metronomic chemotherapy: from the research laboratory into the oncology clinic. PMID- 11863093 TI - Extragonadal retroperitoneal germ cell tumor: evidence of origin in the testis. AB - BACKGROUND: The origin of extragonadal retroperitoneal germ cell tumors remains controversial. Whether they develop primarily in the retroperitoneum or whether they are metastases of a primary testicular tumor has long been debated. PATIENTS AND METHODS: We retrospectively analyzed 26 patients treated as having primary extragonadal retroperitoneal germ cell tumors based upon the findings of testicular palpation by the referring physician. Testicular evaluation was then extended with ultrasonographical and histological examinations. RESULTS: Biopsy of the extragonadal tumor was performed in 25 patients, confirming diagnosis of extragonadal retroperitoneal germ cell tumor. Prior to treatment patients were clinically evaluated by several physicians and the testes were not considered suspicious for testicular cancer. At urological workup, testes were found to be atrophic and/or indurated in 14 (54%) patients, enlarged in one (4%) and unremarkable in 11 (42%). Ultrasound examination of the testes in 20 patients showed pathological findings in all of them. Histology of the testis was available in 25 of 26 patients and revealed active tumor in three, intratubular germ cell neoplasia in four, scar tissue in 12, sclerosis in three, sclerosis and fibrosis in one, and fibrosis alone in two. CONCLUSIONS: So-called primary extragonadal germ cell tumors in the retroperitoneum are very likely a rare or non-existing entity and should be considered as metastases of a viable or burned out testicular cancer until proven otherwise. All of our patients with histologically examined testes had pathological finding, 76% of which were either viable tumor or scars. PMID- 11863094 TI - A 20% dose reduction of the original CISCA/VB regimen allows better tolerance and similar survival rate in disseminated testicular non-seminomatous germ-cell tumors: final results of a phase III randomized trial. AB - BACKGROUND: This prospective randomized clinical trial was designed to compare the efficacy of a low-dose regimen of cisplatin, doxorubicin and cyclophosphamide alternated with vinblastine and bleomycin (CISCA/VB) with the original CISCA/VB regimen in patients with disseminated nonseminomatous germ-cell tumors (NSGCT) and a predicted favorable outcome. PATIENTS AND METHODS: One hundred and twenty five patients with disseminated NSGCT and a predicted favorable outcome according to the M.D. Anderson Cancer Center classification [testicular primary and human chorionic gonadotropin (hCG) serum level <50000 mIU/ml] were randomized to receive the original CISCA/VB regimen (100% dose) or a low-dose CISCA/VB regimen (80% dose). RESULTS: Among the 124 eligible patients, there was no significant difference in the number of patients in the two treatment arms who achieved a complete response to therapy: 53 of 65 patients (82%) on the original CISCA/VB regimen and 53 of 59 patients (90%) on the low-dose CISCA/VB regimen (P = 0.29). Overall, the original CISCA/VB regimen resulted in a significantly higher relative dose intensity (P <0.0001). After a median follow-up of 6.8 years (range 0.37 to 12.94 years), there was no significant difference in disease-free survival (P = 0.87) or in overall survival (P = 0.88) between the two treatment arms. The 5-year overall survival rate was 93.7% [95% confidence interval (CI) 88% to 100%] and 94.1% (95% CI 84% to 100%) in the original CISCA/VB arm and the low-dose CISCA/VB arm, respectively. The 5-year overall survival rate for the entire study population was 98% (95% CI 94% to 100%) and 88% (95% CI 76% to 100%) in the good- and intermediate-prognosis groups, respectively, as defined by the International Germ Cell Consensus Classification Group (IGCCCG). The low-dose CISCA/VB regimen resulted in significantly less neutropenic fever (P <0.001), grade 4 thrombocytopenia (P <0.03) and severe mucositis (P <0.01) than the original CISCA/VB regimen. CONCLUSIONS: CISCA/VB is highly efficient in patients with good or intermediate prognosis NSGCT according to the IGCCCG criteria. Although equivalent antitumor efficacy cannot be claimed, the low-dose CISCA/VB regimen appears to be a better mode of delivery than the original CISCA/VB regimen with respect to toxicity, since survival is comparable. PMID- 11863095 TI - Allogeneic hematopoetic stem-cell transplantation for patients with relapsed or refractory lymphomas: comparison of high-dose conventional conditioning versus fludarabine-based reduced-intensity regimens. AB - BACKGROUND: Allogeneic hematopoetic stem-cell transplantation (alloHSCT) has curative potential for poor risk lymphoma patients due to the graft-versus lymphoma effect. High non-relapse mortality with conventional high-dose conditioning indicates the necessity for less toxic transplant strategies. PATIENTS AND METHODS: Between 1992 and 1999, 25 patients [median age 37 (20-60) years] with relapsed or refractory non-Hodgkin's lymphoma (NHL, n = 20) or Hodgkin's disease (HD, n = 5) received an alloHSCT in our institution. Patients were grafted from HLA matched (17) or mismatched (2) related, or matched unrelated donors (MUD) (6). NHL histological subtypes were lymphoblastic (6), high grade B/T-cell lymphomas (5), follicular (3), mantle cell (2) and CLL, immunocytic, composite lymphoma and panniculitic T-NHL in one patient each. Patients had received a median of four (range three to six) different therapies before alloHSCT, and 10 patients had relapsed after high-dose chemotherapy and autologous (9) or allogeneic (1) HSCT. Remission status prior to allogeneic SCT was CR1 (1), CR2 (1), relapse (11), partial remission (5) or primary refractory induction failure (7). Conventional myeloablative conditioning (cc) regimens contained total body irradiation 12 Gy (5), busulfan 16 mg/kg (7) or BCNU/VP16 (1). Twelve patients received reduced-intensity conditioning (ric) regimens with fludarabine (FLU) plus alkylating agents. Graft-versus-host disease prophylaxis consisted of cyclosporin A +/- prednisone or methotrexate. Six patients also received anti-T-lymphocyte globulin. RESULTS: Twenty-four patients engrafted. Best response after alloHSCT was complete remission in 16 of all patients [64%: 95% confidence interval (CI) 44% to 84%] and in 16 of 22 evaluable patients (73%: 95% CI 53% to 93%), partial remission in three of 25 (12%), and no change in three of 25 (12%) patients. Early death prevented response evaluation in three of 25 patients. Non-relapse mortality was 54% (95% CI 15% to 78%) in patients after cc and 17% (95% CI 0% to 41%) after FLU-based ric (P = 0.03). Six patients died due to progressive disease or relapse. Four patients with HD died, three in complete remission due to non-relapse mortality and one with progressive disease. Eleven of 25 patients are alive with a median follow up of 618 days (range 383 2815), with an overall survival of 44% (95% CI 23% to 65%) at 1 year for all patients, while eight of 12 (67%: 95% CI 35% to 98%) patients are alive after ric compared with three of 13 (23%; 95% CI 0% to 50%) after cc (P <0.02). CONCLUSIONS: AlloHSCT induces high rates of complete remission in advanced lymphoma patients, even when the tumor had relapsed after autologous HSCT. It should be considered earlier as part of the therapeutic options in poor risk patients to avoid non-relapse mortality associated with extensive pretreatment. Our novel reduced conditioning regimens show promising results, especially in heavily pretreated patients, and improve survival after allogeneic transplantation. PMID- 11863096 TI - Peripheral T-cell lymphoma (excluding anaplastic large-cell lymphoma): results from the Non-Hodgkin's Lymphoma Classification Project. AB - BACKGROUND: Peripheral T-cell lymphoma (PTCL) is rare in most parts of the world. Therefore, we have evaluated the 96 cases of PTCL diagnosed within the Non Hodgkin's Lymphoma Classification Project (NHLCP) (1378 cases) for their geographical distribution, pathologic features and diagnostic reliability, as well as clinical presentation and outcome. MATERIALS AND METHODS: Diagnoses of all cases were rendered independently by five experienced hematopathologists based on morphology only, and after introduction of the immunophenotype and clinical data. Divergent diagnoses were jointly discussed and a final consensus diagnosis was established in each case. Reliability of the diagnoses was evaluated statistically, and the clinical features and outcome were analyzed according to the consensus diagnoses. RESULTS: Seven per cent of all non Hodgkin's lymphoma (NHL) cases reviewed were classified as PTCL and the frequency varied from 1.5% to 18.3% in different countries. The interobserver agreement with the consensus diagnosis of PTCL was 86% in the Revised European-American Lymphoma (REAL) classification, but the designation of subtypes was less reliable. Diagnostic reliability improved from 41% to 86% after immunophenotyping, but did not improve further with the addition of detailed clinical data. Clinically, angiocentric nasal lymphoma presented in young females (median age 49 years) at extranodal sites, but with few adverse risk factors, whereas angioimmunoblastic lymphoma presented most often in older males (median age 65 years) at nodal and extranodal sites with numerous risk factors. The 5 year overall and failure-free survivals for patients with PTCL treated with doxorubicin (Adriamycin)-containing regimens were only 26% and 20%, respectively. Both failure-free and overall survival were strongly correlated with the performance status and International Prognostic Index scores at presentation, but differences in survival were not observed between the major histological types. However, within the PTCL 'not otherwise specified' category, but not angioimmunoblastic lymphoma, the number of transformed blasts was prognostically relevant. CONCLUSIONS: PTCLs can be diagnosed reliably by experienced hematopathologists, but immunophenotyping is absolutely necessary. Currently, all types of PTCL should be considered high-grade lymphomas. An increased ability to distinguish T-lymphocyte subsets is needed in order to better subclassify the PTCLs for therapeutic and prognostic purposes. PMID- 11863097 TI - FDG-PET for detection of recurrences from malignant primary bone tumors: comparison with conventional imaging. AB - BACKGROUND: The aim of this study was to assess the diagnostic ability of positron emission tomography using 2-[fluorine-18]fluoro-2-deoxy-D-glucose (FDG PET) in the detection of recurrences from malignant primary bone tumors compared with conventional imaging. PATIENTS AND METHODS: In 27 patients (6 osteosarcomas, 21 Ewing's sarcomas), 41 FDG-PET examinations performed for diagnosis or exclusion of recurrent disease were evaluated. Conventional imaging techniques consisted of magnetic resonance imaging of the primary tumor site, thoracic computed tomography, and Tc-99m methylene diphosphonate bone scintigraphy. The reference methods were the histopathological analysis and/or the clinical and imaging follow-up. RESULTS: In 25 examinations reference methods revealed 52 sites of recurrent disease (local n = 7; distant: osseous n = 22, pulmonary n = 13, soft tissue n = 10). On an examination-based analysis FDG-PET had a sensitivity of 0.96, a specificity of 0.81 and an accuracy of 0.90. Corresponding values for conventional imaging were 1.0, 0.56 and 0.82. CONCLUSIONS: The sensitivity, specificity and accuracy of FDG-PET in the detection of recurrences from osseous sarcomas are high. On an examination-based analysis, FDG-PET had a not significantly lower sensitivity in comparison with conventional imaging. However, FDG-PET showed a small advantage in the detection of osseous and soft tissue recurrences compared with conventional imaging. PMID- 11863098 TI - Faecal occult blood testing (FOBT) as screening for colorectal cancer: the current controversy. PMID- 11863099 TI - Phase I study of twelve-day prolonged infusion of high-dose ifosfamide and doxorubicin as first-line chemotherapy in adult patients with advanced soft tissue sarcomas. AB - PURPOSE: To determine whether a prolonged 12-day continuous infusion allows the administration of high-dose ifosfamide (IFO) with an acceptable toxicity profile when combined with full-dose doxorubicin (Adriamycin; ADM) as first-line chemotherapy in patients with advanced soft tissue sarcomas. PATIENTS AND METHODS: Escalating doses of continuous infusion IFO (8-15 g/m2) given on days 1 to 12 in combination with ADM 75 mg/m2 given on day 8 and prophylactic granulocyte colony-stimulating factor support were administered every 4 weeks to 35 chemonaive patients with advanced soft tissue sarcomas. RESULTS: The maximum tolerated dose was IFO 15 g/m2. Hematological toxicity was the main dose-limiting toxicity and was dose dependent. Furthermore, thrombocytopenia was cumulative. Grade 4 (WHO) neutropenia and thrombocytopenia were recorded in 48% and 14% of courses, respectively. Eight patients experienced febrile neutropenia. A partial response was observed in 16 out of 30 assessable patients [53%, 95% confidence interval (CI) 25-63]; median time to progression was 25 weeks (range 4-91). CONCLUSIONS: This study proved that a prolonged 12-day continuous infusion allows an increase in the total IFO dose that can be safely combined with ADM. A multicentric phase II study by the Italian Sarcoma Group to assess its antitumor activity is currently ongoing in patients with advanced soft tissue sarcomas. PMID- 11863100 TI - Elevated alpha-foetoprotein and hepatic metastases--it's not always what it seems! AB - Gastric cancers that secrete alpha-foetoprotein (AFP) are a rare but well defined entity. These tumours tend to be aggressive with a poor prognosis. Recognition and identification of this group of patients is important in influencing prognosis, treatment options and further clinical research into this entity. We report three cases of gastric cancers associated with elevated AFP. This entity should be considered in patients with hepatic lesions and an elevated AFP without risk factors for hepatocellular carcinoma or in patients with associated gastric symptoms. PMID- 11863101 TI - A paraneoplastic syndrome mimicking extrauterine pregnancy. AB - We report on a 30-year-old female patient with a beta-human chorionic gonadotropin (beta-HCG)-producing lung tumour. Abdominal discomfort and vaginal bleeding were the presenting symptoms and, in conjunction with elevated beta-HCG levels, initially led to the diagnosis of extrauterine pregnancy. Bilateral ovarian cysts were detected on further diagnostic workup. Ultimately, a chest X ray revealed a lung tumour. The paraneoplastic symptoms were completely reversible after resection of the lung lesion, and the ovarian cysts disappeared. PMID- 11863102 TI - Massive cavitation of solid pulmonary metastatic lesions in a breast cancer patient: a case report. AB - A 40-year-old female with metastatic breast cancer developed multiple lung nodules some of them with cavitations. Following treatment with Taxol/Herceptin most of the lesions disappeared and in many cavity lesions appeared. There was no further change in the appearance of lung lesions. PMID- 11863103 TI - Reply to the article "The AJCC staging proposal for cutaneous melanoma: comments by the EORTC Melanoma Group", by D. J. Ruiter et al. (Ann Oncol 2001; 12: 9-11). American Joint Committee on Cancer. PMID- 11863104 TI - Regulation of cigarette emissions. PMID- 11863105 TI - A systematic review of the role of pulmonary irradiation in the management of primary bone tumours. AB - INTRODUCTION: Adjuvant therapy in osteosarcoma (OS) and Ewing's sarcoma (ES) is primarily directed towards treatment of subclinical lung disease. Before the advent of modern intensive chemotherapy, lung irradiation was the only available adjuvant treatment. It has proven biological activity and low morbidity. There is, however, a wide variation in its application between centres. This systematic review aims to define the evidence to support the use of lung irradiation in these diseases. DESIGN: A review of trials published between 1966 and 2000 was undertaken to determine the evidence for the use of pulmonary irradiation in OS and ES. RESULTS: Several small series of prophylactic lung irradiation (PLI) have been reported, most from over 20 years ago. These studies support the theoretical basis for the use of PLI in both OS and ES. Few randomised studies have been performed which include PLI. In OS, studies demonstrated a trend in favour of PLI compared with no adjuvant treatment and, subsequently, a level of benefit similar to that achieved with chemotherapy, but no additive effect. No studies have used PLI in addition to current standard chemotherapy regimens, or evaluated its use after successful metastatectomy. In ES, only one randomised study has addressed the role of PLI, in a comparison with vincristine, actinomycin D and cyclophosphamide combination chemotherapy with or without doxorubicin. Prolonged follow-up favoured four-drug chemotherapy. Retrospective reports from large cooperative groups suggest that the addition of whole-lung radiotherapy (WLRT) improves outcome in ES patients presenting with pulmonary metastases. However, there are no randomised study data to support this. CONCLUSIONS: Further randomised studies are necessary to clarify the role of PLI in addition to current standard chemotherapy regimens, or its use after successful metastasectomy in patients with OS. In patients with localised ES adjuvant chemotherapy appears to be superior to PLI alone, while there is little evidence to support treatment with WLRT in patients who present with pulmonary metastases. PMID- 11863107 TI - Faecal occult blood testing for colorectal cancer. PMID- 11863106 TI - Fecal occult blood screening for colorectal cancer: open issues. AB - Data from seven case-control and--mainly--three randomized clinical trials consistently indicate that biennial fecal occult blood screening (FOBT) can reduce colorectal cancer (CRC) mortality by approximately 20% after 10-18 years. The reduction may be greater in compliant subjects. In the long-term, incidence also appears to be reduced. There are suggestions that the effect of annual screening may be greater, although data are inadequate to quantify the potential advantages of annual versus biennial screening. The issue of the effectiveness of FOBT in the general population and, more important, of comparative cost effectiveness with other possible screening tests for CRC, however, remain open to discussion. PMID- 11863109 TI - Hemoccult should no longer be used for the screening of colorectal cancer. PMID- 11863108 TI - Fecal occult blood testing as a screening procedure for colorectal cancer. PMID- 11863110 TI - Is colorectal cancer screening by fecal occult blood feasible? PMID- 11863111 TI - Fecal occult blood test for colorectal cancer screening. PMID- 11863112 TI - Should organised faecal occult blood test screening be established? PMID- 11863113 TI - Fecal occult blood testing for colorectal cancer: a perspective. AB - Colorectal cancer is an important health problem in western countries. Early detection of colorectal cancer reduces mortality. The best evidence for the effectiveness of screening for colorectal cancer is with annual or biennial fecal occult blood testing. While the benefit of fecal occult blood testing is small in absolute terms, the incremental cost-effectiveness of this screening strategy appears acceptable. Combining fecal occult blood testing with periodic flexible sigmoidoscopy or replacing it altogether with infrequent colonoscopy are theoretically attractive screening strategies, but the incremental costs and effectiveness of these more intensive screening strategies have not been well defined. Whether and how to implement population-based screening for colorectal cancer depends largely on available resources. PMID- 11863114 TI - Cooperative inhibitory effect of ZD1839 (Iressa) in combination with trastuzumab (Herceptin) on human breast cancer cell growth. AB - BACKGROUND: Co-expression of the epidermal growth factor receptor (EGFR) and of ErbB-2 is found in a subset of primary human breast cancer. MATERIALS AND METHODS: The antiproliferative effects of anti-EGFR and anti-ErbB-2 agents were evaluated using a monolayer assay. The effects of these agents on the activation of EGFR, ErbB-2, AKT and p42/p44 MAP kinases (MAPK) were investigated by western blot analysis. RESULTS: We found that both ZD1839 (Iressa), a specific EGFR tyrosine kinase inhibitor, and trastuzumab (Herceptin) (TRA), a humanized anti ErbB-2 monoclonal antibody, were able to inhibit the growth of SK-Br-3 and BT-474 breast carcinoma cells, which express both EGFR and ErbB-2. Treatment of breast carcinoma cells with a combination of ZD1839 and TRA resulted in a synergistic inhibitory effect. Treatment of SK-Br-3 cells with ZD1839 produced a significant, dose-dependent reduction of the tyrosine phosphorylation of both EGFR and ErbB-2. Phosphorylation of MAPK and AKT were significantly reduced in SK-Br-3 cells following treatment with ZD1839, whereas treatment with TRA produced a reduction of AKT but not MAPK phosphorylation. Finally, treatment with ZD1839, but not with TRA, produced a significant increase in fragmented DNA in breast carcinoma cells. However, a more pronounced increase in the levels of fragmented DNA was observed following combined treatment with ZD1839 and TRA. CONCLUSIONS: These data suggest that combined treatment with drugs that target EGFR and ErbB-2 might result in an efficient inhibition of tumor growth in those breast carcinoma patients whose tumors co-express both receptors. PMID- 11863115 TI - Low-dose oral methotrexate and cyclophosphamide in metastatic breast cancer: antitumor activity and correlation with vascular endothelial growth factor levels. AB - BACKGROUND: Anticancer chemotherapy is thought to be effective by means of direct cytotoxicity on tumor cells. Alternative mechanisms of efficacy have been ascribed to several common anticancer agents, including cyclophosphamide (CTX), methotrexate (MTX), anthracyclines and taxanes, postulating an antiangiogenic activity. PATIENTS AND METHODS: We evaluated the clinical efficacy and impact on serum vascular endothelial growth factor (VEGF) levels of low-dose oral MTX and CTX in patients with metastatic breast cancer. MTX was administered 2.5 mg bd on days 1 and 2 each week and CTX 50 mg/day administered continuously. RESULTS: Sixty-four patients were enrolled, 63 were evaluable: Eastern Cooperative Oncology Group (ECOG) performance status 0-1, > or =2 sites of metastatic disease (n = 50 patients), progressive disease at study entry (n = 51), 1 regimen for metastatic disease (n = 32) and > or =2 regimens (n = 20). Among the 63 evaluable patients, there were two complete remissions (CR), 10 partial remissions (PR) for an overall response rate of 19.0% (95% CI 10.2% to 30.9%) and an overall clinical benefit (CR+ PR+ stable disease >24 weeks) of 31.7% (95% CI 20.6% to 44.7%). Grade > or =2 leucopenia was registered in only 13 patients. The median serum VEGF level for the subgroup of patients on treatment for at least 2 months decreased with treatment from 315 pg/ml (95% CI 245 to 435) at baseline to 248 pg/ml (95% CI 205 to 311) at 2 months (P <0.001). Both responders and non responders showed similar reductions in serum VEGF (P = 0.78). After 6 months patients still on treatment had a median VEGF level of 195 pg/ml (95% CI 96 to 355), which was significantly lower than the median baseline values (P = 0.001). CONCLUSIONS: Continuously low-dose CTX and MTX is minimally toxic and effective in heavily pretreated breast cancer patients. A drop in VEGF was associated with the treatment and so alternative hypotheses, other than that of direct toxicity on tumor cells, must be favored when trying to explain the anticancer effect. PMID- 11863116 TI - Combined anti-EGF receptor and anti-HER2 receptor therapy in breast cancer: a promising strategy ready for clinical testing. PMID- 11863117 TI - A double-blind placebo-controlled randomized phase III trial of 5-fluorouracil and leucovorin, plus or minus trimetrexate, in previously untreated patients with advanced colorectal cancer. AB - BACKGROUND: Trimetrexate (TMTX) biochemically modulates 5-fluorouracil (5-FU) and leucovorin (LCV). Two phase II trials demonstrated promising activity for TMTX/5 FU/LCV in patients with untreated advanced colorectal cancer (ACC). This trial was designed to demonstrate the safety and efficacy of TMTX/5-FU/LCV as first line treatment in ACC. PATIENTS AND METHODS: Eligible patients with ACC were randomized in double-blind fashion to receive placebo or TMTX (110 mg/m2) intravenously (i.v.) followed 24 h later by i.v. LCV 200 mg/m2, and 5-FU 500 mg/m2 plus oral LCV rescue. Both schedules were given weekly for 6 weeks every 8 weeks. Patients were evaluated for progression-free survival (PFS), overall survival (OS), tumor response, quality of life (QoL) and toxicity. RESULTS: A total of 382 eligible patients were randomized. Significant toxicities were noted more frequently with TMTX/5-FU/LCV. Diarrhea was the most common grade 3 or 4 side-effect (41% and 28% on the TMTX and placebo arms, respectively). QoL scores and response rates did not differ between treatment arms. PFS was 5.3 months and 4.4 months in the TMTX and placebo arms, respectively (P = 0.77; Wilcoxon). OS was 15.8 months and 16.8 months, respectively (P = 0.73; Wilcoxon). CONCLUSIONS: The addition of TMTX to a weekly regimen of 5-FU/LCV worsened grade 3 or 4 diarrhea. The inclusion of TMTX did not yield any significant improvements in response rate, PFS or OS. PMID- 11863118 TI - Maintenance daily oral etoposide versus no further therapy following induction chemotherapy with etoposide plus ifosfamide plus cisplatin in extensive small cell lung cancer: a Hoosier Oncology Group randomized study. AB - BACKGROUND: We performed this phase III study to determine whether the addition of 3 months of oral etoposide in non-progressing patients with extensive small cell lung cancer (SCLC) treated with four cycles of etoposide plus ifosfamide plus cisplatin (VIP) improves progression-free survival (PFS) or overall survival. PATIENTS AND METHODS: Patients with extensive SCLC with a Karnofsky performance score (KPS) > or =50, adequate renal function and bone marrow reserve were eligible. Patients with CNS metastasis were eligible and received concurrent whole-brain radiotherapy. All patients received etoposide 75 mg/m2, ifosfamide 1.2 g/m2 and cisplatin 20 mg/m2 intravenously on days 1-4 every 3 weeks for four cycles. Non-progressing patients were randomized to oral etoposide 50 mg/m2 for 21 consecutive days every 4 weeks for three courses versus no further therapy until progression. RESULTS: From September 1993 to June 1998, 233 patients were entered and treated with VIP with 144 non-progressing patients subsequently randomized to oral etoposide (n = 72) or observation (n = 72). Minimum follow up for all patients is 2 years. Toxicity with oral etoposide was mild. There was an improvement in median PFS favoring the maintenance arm of 8.23 versus 6.5 months (P = 0.0018). There was a trend towards an improvement in median (12.2 versus 11.2 months), 1-year (51.4% versus 40.3%), 2-year (16.7% versus 6.9%) and 3-year (9.1% versus 1.9%) survival (P = 0.0704) favoring the maintenance arm. CONCLUSIONS: Three months of oral etoposide in non-progressing patients with extensive SCLC was associated with a significant improvement in PFS and a trend towards improved overall survival. PMID- 11863119 TI - Comparative effects of plasmapheresis and intravenous cyclophosphamide on urinary podocyte excretion in patients with proliferative Lupus nephritis. AB - Intravenous cyclophosphamide (IVC) in combination with steroids is standard therapy for Lupus nephritis. Reduction of autoantibodies and circulating immune complexes can be used in the treatment of autoimmune diseases. The aim of the present study was to compare the effects of IVC pulse therapy and double filtration plasmapheresis (DFPP) on proteinuria and urinary excretion of podocytes in adult patients with diffuse proliferative Lupus nephritis (DPLN). Twenty patients were randomly assigned to two groups. Group A (n = 10) was treated with IVC (0.75 - 1.0 g/m2 body surface area) pulse therapy, given as boluses once a month for 6 consecutive months, combined with oral corticosteroid (up to 1 mg/kg/day) administration. Group B (n = 10) was treated with a combination of DFPP (performed 1-2 times weekly) and corticosteroid (up to I mg/kg/ day). The total average number of treatments was 8.4 and the therapeutic efficacies were evaluated after 6 months. Twenty healthy individuals participated as a control group. Urinary podocytes were examined by immunofluorescence with monoclonal antibodies against podocalyxin. Both Group A and Group B reduced proteinuria (p < 0.001) as well as the number of urinary podocytes (p < 0.001). Differences between the 2 treatment outcomes were not statistically significant. Cyclophosphamide pulse therapy and DFPP may be similarly effective in the treatment of podocyte injury in patients with DPLN. PMID- 11863120 TI - Lupus nephritis: a retrospective review of 78 cases from a single center. AB - Lupus nephritis (LN) is a frequent and serious manifestation of systemic lupus erythematosus. However, the outcome has progressively improved in the last 3 decades and this was due to more efficient and early treatment of LN and comorbid situations. The aim of our study was to analyze our experience and outcome in LN, to evaluate clinicopathologic and clinicolaboratory correlations and to search for risk factors for renal and patient survival. PATIENTS AND METHODS: We conducted a retrospective study of 78 patients with biopsy-proven LN. RESULTS: Acute renal failure and subnephrotic proteinuria with microhematuria occurred each one in 39.7% of the patients, nephrotic syndrome in 16.7% and nephritic syndrome in 3.8%. The mean serum creatinine at presentation was 1.45 +/- 1.03 mg/dl and the creatinine clearance was 68.2 +/- 40.3 ml/min. Class IV LN existed in 71.8%, Class III in 20.5%, Class V in 6.4% and Class VI in 1.3%. The treatment included steroids and cytotoxic agents in 87.5% of the patients with proliferative LN. Hypertension, serum creatinine and acute renal failure at presentation, as well as significant chronicity on renal biopsy, were significantly correlated with the progression to chronic renal failure in our population. Males were more prone to develop renal flares. 3.8% of the patients died, 9% lost their renal function, 26.9% are in remission, 33.3% still have subnephrotic proteinuria and microhematuria, 7.7% have nephrotic syndrome and 19.2% have chronic renal failure. The mean global follow-up was 102 +/- 74 months and 96.2% of the patients survived. The actuarial renal survival was 96.1% in the first year; 89.9% at 5 years; and 83.7% at 10 and 20 years. CONCLUSION: We can say that hypertension, serum creatinine and acute renal failure at the onset and significant chronicity on renal biopsy, proved to be risk factors for chronic renal failure in our study population. Male gender was a risk factor for renal flares. The achieved global outcome can be considered a good result. PMID- 11863121 TI - Minimal change disease in systemic lupus erythematosus. AB - We report the clinical and pathologic findings in 7 patients with systemic lupus erythematosus and minimal change disease. All 7 patients presented with full nephrotic syndrome including peripheral edema, nephrotic range proteinuria (mean 9.6 g/day), and hypoalbuminemia (mean 1.8 g/dl). In all cases, renal biopsy revealed diffuse foot process effacement in the absence of significant peripheral capillary wall immune deposits, findings consistent with minimal-change disease. In addition, 5 cases displayed mesangial electron-dense deposits, with or without associated mesangial proliferation, consistent with underlying lupus nephritis class II. In all cases, steroid therapy induced a rapid remission of nephrotic syndrome. Minimal change disease is an underrecognized and readily reversible form of nephrotic syndrome in systemic lupus erythematosus. Because it may occur superimposed on mild mesangial proliferative lupus nephritis, this entity may be misinterpreted as an atypical presentation of lupus nephritis class II. Proper recognition of this entity requires careful integration of the renal biopsy immunofluorescence and electron microscopic findings. PMID- 11863122 TI - Renal function in children with sickle cell anemia. AB - BACKGROUND: Patients with sickle cell anemia have various forms of renal dysfunction. SUBJECTS, MATERIALS AND METHODS: The purpose of this study is to demonstrate the abnormalities of HbSS patients' renal function in childhood. Renal function studies were performed in 55 patients with homozygote sickle cell anemia and compared with 13 healthy children. The blood and timed urine samples were obtained for hematological and biochemical determinations. RESULTS: Mean serum creatinine, sodium, phosphorus and calcium levels were not statistically different between patients and controls. Mean serum potassium and uric acid levels were significantly higher in patients than in controls. Mean tubular phosphate reabsorption (p < 0.001) and fractional excretion of potassium (p < 0.05) were lower in patients than in the control. There were no significant differences in fractional excretion of sodium and uric acid between patients and controls. Patients had significantly higher urine pH and significantly lower specific gravity and osmolality than controls. Also, there were no significant differences in urinary protein/ creatinine, urinary N-acetyl-beta-D glucosaminidase/creatinine and urinary malondialdehyde/creatinine between patients and controls. CONCLUSION: Thus, significant proximal tubular dysfunction is not a common feature but distal tubular abnormality is the most consistent renal functional derangement of patients with SCA in childhood. PMID- 11863125 TI - Renal involvement in sarcoidosis--a report of 6 cases. AB - This report concerns 6 patients with renal involvement in sarcoidosis. Two of the patients had no clinical symptoms at all. In 3 patients, no extrarenal organ manifestation was found. All 6 patients had elevated levels of serum creatinine, 2 were hypercalcemic. Five patients manifested with mild proteinuria, but in none of the cases was a nephritic sediment with erythrocytes found. Kidney biopsies in 5 patients showed epitheloid cell granulomatous interstitial nephritis, and 1 patient presented with nephrocalcinosis. All patients were treated with corticosteroids. The serum creatinine levels decreased significantly in 4 patients (> 50% decrease), and slightly in 2 patients, elevated serum calcium levels were normalized. Thus, even in the absence of other organ manifestations, sarcoidosis can be the cause of renal insufficiency, and it responds well to corticosteroid treatment. These patients demonstrate the importance of kidney biopsy in the unexplained deterioration of renal function. PMID- 11863124 TI - Reduction in erythropoietin doses by the use of chronic intravenous iron supplementation in iron-replete hemodialysis patients. AB - BACKGROUND: Iron deficiency is the most common cause of suboptimal response to recombinant human erythropoietin (rHuEPO) in chronic hemodialysis (HD) patients. Iron supply can correct this situation, however, optimal dosage, route of administration, and monitoring of iron status during rHuEPO therapy in maintenance HD patients remains controversial. METHODS: We conducted a 12-month intravenous iron substitution trial in 149 iron-replete chronic HD patients receiving subcutaneous rHuEPO therapy. The available iron pool was maintained with 100 mg iron every 2 weeks or 1 month depending on serum ferritin and transferrin saturation levels, the rHuEPO dosage titrated depending on hematocrit (Hct) levels. RESULTS: After 12-month protocol, the Hct increased (28.7 +/- 4.1 vs 27.7 +/- 2.6, p = 0.003), rHuEPO requirement reduced 25% (46.1 +/- 28.9 vs 61.5 +/- 67.8 U/kg/week, p = 0.006), serum ferritin increased (1,383 +/- 727 vs 930 +/- 857 ng/ml, p < 0.001), so did the transferrin saturation (36.1 +/- 12.7 vs 27.5 +/- 12.8%, p < 0.001). The serum albumin decreased slightly but reached statistical significance (4.1 +/- 0.48 vs 4.2 +/- 0.36 g/dl, p = 0.006), so did the cholesterol levels (166 +/- 41 vs 173 +/- 38 mg/dl, p = 0.044) and pre dialysis creatinine (11.3 +/- 2.3 vs 11.5 +/- 2.4 mg/dl, p = 0.015). Besides, the iPTH levels did not interfere with the rHuEPO dosage reduction and Hct increment in our patients. CONCLUSION: We conclude that maintaining high levels of serum ferritin and transferrin saturation could further reduce the requirement of rHuEPO in chronic HD patients, but the long-term effect of iron overloading to patients' nutritional status must be further evaluated in contrast to the economic saving. PMID- 11863123 TI - Calcium-sensing receptor gene polymorphism affects the parathyroid response to moderate hypercalcemic suppression in patients with end-stage renal disease. AB - AIM: The basic mechanism of secondary hyperparathyroidism is still unclear, but a change in Ca2+ sensing by parathyroid cells is possibly involved in this uremic complication. A rightward shift of the calcium set-point and an increase of the minimum secretion rate have been found in secondary hyperparathyroidism, indicating abnormal calcium sensing. METHODS: We evaluated the effect of calcium sensing receptor (CaR) gene polymorphism (codon G990R) on the response of the parathyroid gland to moderate hypercalcemic suppression in 77 ESRD patients on regular hemodialysis (HD using 2.5 mEq/l Ca2+ dialysate). All patients underwent an HD session with 3.0 mEq/l Ca2+ dialysate to suppress parathyroid hormone (PTH). Then we investigated the effect of CaR gene polymorphism on the parathyroid response to hypercalcemic stimulation. RESULTS: Patients were divided into 3 groups on the basis of genotype (GG = 33 patients (42.9%), GR = 39 patients (50.6%), RR = 5 patients (6.5%)). Baseline intact PTH levels in patients without the R allele were not significantly different from those in patients with the R allele (GG group, 181.4 +/- 31.1 pg/ml vs. GR and RR groups, 230 +/- 51.2 pg/ml: mean +/- SEM). The significant effect of moderate hypercalcemic suppression on the intact PTH level was observed in the GG group (p < 0.01) but not in the GR and RR groups, despite the identical increase in Ca2+. CONCLUSION: Our results suggest that CaR gene polymorphism (codon G990R) influences the responsiveness of the parathyroid gland to changes of extracellular Ca2+ in ESRD patients. The glands of patients with the GG genotype of the CaR gene may be more sensitive to extracellular Ca2+ changes. PMID- 11863127 TI - Hyperhomocysteinemia, low folate status, homozygous C677T mutation of the methylene tetrahydrofolate reductase and renal arterial thrombosis. AB - We report a renal artery thrombosis in a 42-year-old man. Fasting homocysteinemia was at 23 micromol/l 3 months later and at 33 pmol/l 5 months after the vascular event. A homozygous C677T MTHFR was found with low folate status. Active smoking may also have contributed to the pathogenesis of renal arterial thrombosis. The other causes of thrombophilia were ruled out. Homocysteine lowering treament was started: homocysteine normalized at 10.6 micromol/l. There was no recurrence of vascular event within 18 months. We propose mild or moderate hyperhomocysteinemia triggered by low folate status in patients with homozygous C677T MTHFR as a cause of renal arterial thrombosis. PMID- 11863126 TI - Interstitial nephritis, acute renal failure in a patient with non-fulminant hepatitis A infection. AB - This is the first report from Norway of a patient with interstitial nephritis and renal failure due to non-fulminant hepatitis A virus (HAV) infection. HAV infection was confirmed by positive anti-HAV IgM serology. All tests for other virus infections were negative. At admittance serum creatinine (s-Creat) and blood urea nitrogen (BUN) concentration were 539 microlmol/l and 32.6 mmol/l increasing the following days to 890 micromol/l and 39.9 mmol/l, respectively. Nine courses of hemodialysis had to be given. Kidney biopsy specimen showed interstitial edema, lymphocytic cell infiltration and acute tubular injury with normal glomeruli. Examination with immunohistochemistry was negative. In contrast to the findings associated with HBV and HCV infection in which glomerular disease is predominantly found, the HAV infection in our patient was associated with interstitial nephritis and acute tubular necrosis. The prognosis of the renal failure due to HAV infection was good although the recovery was substantially delayed. PMID- 11863128 TI - Necrotic skin lesions in a dialysis patient: a multifactorial entity. AB - A female dialysis patient with a consistently high serum calcium phosphate product presented with large necrotic skin lesions with ulcers. The clinical course was highly suggestive of calciphylaxis. Parathyroidectomy was followed by the healing of the lesions. New skin lesions appeared following relapse of hyperparathyroidism. Her clinical records included a long past of hypertension, which was the cause of her renal failure. She had a limited walking range and previously had presented bilateral ulcers of vascular origin. This case presents a type of lesion which bears a serious prognosis in dialysis patients. The clinical context and the presentation of the lesions are compatible with multiple etiology: vascular lesions and calciphylaxis. The documented longitudinal follow up illustrates the importance of treating the different factors known to participate in the appearance of skin lesions in dialysis patients. Particularly, it stresses the benefit of performing parathyroidectomy, even if the parathyroid hormone level is not in the range normally accepted as requiring surgical removal of parathyroid glands. PMID- 11863129 TI - Is continuous veno-venous hemofiltration for acetaminophen-induced acute liver and renal failure worthwhile? AB - We describe a patient with acute liver and renal failure secondary to acetaminophen and chronic alcohol abuse who was treated aggressively with oral acetylcysteine, continuous renal replacement therapy, glucose and branched-chain amino acid intravenous feeding and ventilatory support. The patient had a predicted mortality of > 95% without liver transplantation however, with intensive ventilatory, renal, and nutritional support he made a complete recovery. We discuss the benefits of aggressive supportive therapy and suggest that continuous renal replacement therapy may allow gentle fluid removal, excellent control of cerebral edema and intravenous feeding that may favorably influence prognosis. PMID- 11863130 TI - Renal involvement in a patient with idiopathic hypereosinophilic syndrome. PMID- 11863131 TI - Lupus nephritis: a clinical review for practicing nephrologists. AB - The renal manifestations in systemic lupus erythematosus (SLE) are protean and difficult to categorize into clinical syndromes and histologic classes. Lupus nephritis is frequently unrecognized until full-blown nephritic and/or nephrotic syndrome with renal failure emerge. Epidemiologically, approximately one third of SLE patients from unselected populations have renal involvement early during the disease. Most renal abnormalities emerge within the first few years of SLE diagnosis. Currently, most nephrologists agree that an early renal biopsy is worthwhile in those SLE patients with abnormal urinalysis and/or reduced renal function. First, it provides a histologic categorization of the glomerulonephritis as well as an assessment of the degree of activity and chronicity. Second, it provides vital prognostic information. Third, it is beneficial in planning a more rational therapy with or without potentially toxic immunosuppressive agents. Over the last 3 decades, many controlled clinical trials for treatment of lupus nephritis have been completed with a few therapeutic immunosuppressive regimens. Among those agents used. cyclophosphamide and azathioprine provide a reduction of morbidity in those patients afflicted with proliferative forms of lupus glomerulonephritis. A new immunosuppressive agent, mycophenolate mofetil, is being studied for treatment of proliferative forms of lupus glomerulonephritis in a controlled clinical trial at our institution. Immunosuppressive agents and the availability of dialysis and transplantation have improved the survival of patients with lupus nephritis, in particular those with proliferative forms. PMID- 11863132 TI - Applying learning theories to develop teaching strategies for the critical care nurse. Don't limit yourself to the formal classroom lecture. AB - Learning, as defined by Alspach, is "a change in cognitive, psychomotor, and/or affective behaviors." The teaching strategies reviewed in this article have focused on ones that can affect all three learner behaviors if carefully planned and executed by the instructor. It is also key to provide the content in a manner that will appeal to the autonomy and self-direction of the adult learner, keeping in mind the importance of relating new information to previously learned material. Realizing that learners have different learning styles, the instructor also should assess learning styles and vary teaching methods accordingly. Incorporating some of the learner assessments and teaching strategies discussed here can be a change for both the learner and instructor, but it is consistent with modern learning theory where the focus is on the learner. PMID- 11863133 TI - Education, training, and use of unlicensed assistive personnel in critical care. AB - The integration of the role of UAP in critical care requires a long-term commitment to patient centered care by the professional nursing staff. Standardized education and training of professional nurses and UAP, support for nurses who orient UAP, support for the PCTs who precept, and a supporting infrastructure promote integration of UAP in critical care with positive outcomes. The education of UAP does not conclude with orientation. Education, training, and competency revalidation should be ongoing to broaden individuals' knowledge base and enhance skills. The monitoring and evaluation of programs with timely modifications will support or enhance effectiveness of the programs. PMID- 11863134 TI - Mentorship as a teaching strategy. AB - Nursing is facing challenges perhaps unparalleled in its history. As we face the opportunities of the future, mentors play a more important role than ever. Mentors have "provided inspiration, support, and encouragement during high and low points of my development." "have forever changed the course of our practice," taught through "her commitment to the advancement of her students and colleagues, her gentle but persistent encouragement to grow, and her generosity in providing pivotal opportunities," "taught me three lessons: caring gets results, family comes first, and passionate commitment is contagious," offers a potential buoy in the sea of change in health care, and possibly enhances clinical outcomes. Mentors can be found in your boss, teacher, spouse, friend, colleague, or peer. Mentorship is a gift between two people and must be given and accepted as such. Not everyone should act as a mentor; the relationship cannot occur and develop when there is no desire to share. Nursing is about learning and teaching. "The spirit of the nursing profession dies when it is reduced to a set of abstract theories, legal requirements, and expert skills. These are the results, not the goals, of scholarship and leadership. Scholarly endeavors always occur amidst communities of learners engaged in being better practitioners of their discipline." Mentors are the leaders amidst the community of nursing. PMID- 11863135 TI - Beyond the essentials. Teaching critical care to undergraduate and graduate students in nursing. AB - With more new graduates entering the critical care setting and with the evolving role of the advanced practice nurse, teaching critical care content to students in nursing is a challenge for faculty. The burden to educate undergraduate novices and advanced practice nurses to care for the complex critically ill patient is costly for health care agencies. To help prepare the generalist to enter a world of specialty, precepted critical care experiences, faculty/critical care nurse collaboration, critical care simulation experiences, a computerized entry-level critical care examination, and a National Fellowship critical care registry are suggested. Teaching strategies for the advanced practice nurse beyond the American Association of Colleges of Nursing's "essentials" document include consideration of expanded science education, simulation for psychomotor skills and diagnostic reasoning cases, and interdisciplinary education for bioethics and business courses. Although such controversies as preparing the generalist for a specialty and the availability of preceptors continue to be debated, the future of critical care education is exciting and challenging. As technology continues to evolve, and the consumer in a market-driven society becomes more involved and inquisitive, the need for caring remains. That is the joy of teaching critical care nursing. PMID- 11863136 TI - Orientation to the critical care unit. The value of preceptor programs. AB - Effective orientation is vital to the critical care team in providing quality patient care in today's health care environment. Preceptorship is the ideal method of providing orientation, and preceptors are key to effective implementation of the program. With limits to the resources available today, modifications to preceptorship can improve the frequency of providing quality orientation. PMID- 11863137 TI - Dealing with resistant learners. Educator strategies. AB - Resistant learners are one of the many challenges critical care educators face. The educator must be aware of the elements that contribute to learner resistance and work effectively to eliminate them. Efforts should be made to design programs that are outcome focused and provide direct benefit to each learner. When resistance presents, the educator must search for ways to secure the resisters' buy-in or minimize their impact. Managing resistant learners is never easy, but when the educator is armed with the right tools, their negative impact can be reduced or eliminated. PMID- 11863138 TI - The critical care consortium. Maximizing continuing education dollars. AB - Facing the current challenges in health care delivery and tremendous changes predicted for the new millennium, educators must consider nontraditional approaches to meet the learning needs of critical care nurses. The education consortium, as described in this article, should be considered as one possible approach. Consortiums have the potential to enhance program quality, reduce costs, promote positive participant interaction, and strengthen community spirit. The consortium approach is not a panacea, but it has proven to be a successful model for critical care education. PMID- 11863139 TI - Two successful models for preparing competent critical care nurses. The Parkland Health and Hospital System Critical Care and Trauma Nurse Internship and Critical Care Residency. AB - Part of the mission statement of Parkland Health and Hospital System involves participating in educational programs dedicated to the art and science of caring for the sick and injured, the promotion of wellness, and the delivery of health services. The concept of the Nurse Internship and Nurse Residency fits well in the framework of this hospital. The continued support of these programs from the PHHS administration is visible evidence of the institution's commitment to excellence. Together these programs provide a continual supply of competent critical care practitioners to meet the never-ending demand in this large county facility as well as opportunities for new graduates to begin the exciting and rewarding journey into critical care nursing. PMID- 11863140 TI - Clinician involvement in critical care research. AB - Clinician involvement in critical care research will be essential if we are ever to achieve optimal patient outcomes. Educators and unit leadership play a pivotal role in facilitating, mentoring, and coaching clinicians both conduct research on important issues or problems in critical care and to ensure that practice routines are based on the latest research findings. PMID- 11863141 TI - Cultural and ethnic diversity. Unique challenges in critical care education. AB - Cultural and ethnic diversity are facts of life in modern America. This diversity adds greatly to a society's growth and development much as herbs and spices make the difference between a really great soup and a bowl of hot colored water. Yet, as any great cook knows, one has to work with the spices to discover how to blend and mix them. The critical care environment intensifies the tendency for cultures to collide and for education to be stressful and unsuccessful. An educator who is able to learn about other cultures, use flexibility and creativity in teaching methods, and be sensitive to the needs of special groups will find this cultural diversity to be part of the spice of successful educational offerings and a rewarding career. PMID- 11863142 TI - Computer-assisted education for critical care nurses. AB - Technology is changing rapidly and health care is just beinnng to see the wave of technological advances. Computer-assisted educational software is available for many topics and in many media. Educators and learners need to explore these media and determine how they can best fit into a total learning experience. Computers should be used to enhance education and training, rather than replace the human instructor. The latest software and hardware are interesting to learners, but technology needs to be weighed along with outcomes of education. Over the next 10 years, many of the materials we use today for critical care education will be replaced with more advanced technologies. Subject matter experts should work with computer experts to design and improve computer-assisted technology. In addition, all educators should assess the return on investment of these newer technologies before embracing them. PMID- 11863143 TI - Teaching in crisis. Patient and family education in critical care. AB - Although the critical care setting is not always a positive teaching environment, it is possible to achieve the goal of optimal patient and family education. The critical care nurse must understand the unique learning needs of patients and families who are experiencing a life crisis a recognize that there are substantial obstacles to overcome to educate in this setting. In addition, it takes experience and resources to develop the teaching skills of the bedside nurse, so that those teachable moments are easily recognized and suitably used to give patients and family members valuable information in small doses. The advanced practice nurse is an essential nursing resource who can spearhead the development of teaching skills for all members of the health care team. In addition, the advanced practice nurse is a clinical expert who can assess the educational needs of patients and their families and provide more detailed and individualized health information from a different perspective. Achieving good patient and family education outcomes is possible when patient care continuity is a priority and the advanced practice nurse is an active part of the nursing team. Exploring the use of new technologies and resources to meet patient and family education needs is absolutely necessary. As hospitals continue to evolve and react to the financial demands placed on them, nursing leadership and critical care nurses will need to articulate clearly all of the essential components of patient care, including patient and family education. In keeping with the rich nursing tradition of patient and family education, critical care nurses and advanced practice nurses have the opportunity to demonstrate their unique teaching skills and continue to promote health education as a priority of patient care. PMID- 11863144 TI - Using simulation to teach critical thinking skills. You can't just throw the book at them. AB - Critical thinking is essential in critical care practice. How to define, measure, and teach thinking are subjects of debate with no clearly proven method for doing so. Simulation as a teaching strategy appears to hold great promise for teaching critical thinking. Sophisticated simulators allow real patient situations to be replicated in a safe environment for practice and learning. Research is needed to support this theory. Dr. Gaba, a pioneer in the use of simulation, said "No industry in which human lives depend on the skilled performance of responsible operators has waited for unequivocal proof of the benefit of simulation before embracing it." Embracing simulation in academic and clinical education is appropriate, but efforts must continue to evaluate this strategy. PMID- 11863145 TI - Illicit drugs for toothache. PMID- 11863146 TI - Stigma knows no bounds. PMID- 11863147 TI - Private dentistry pricing. PMID- 11863148 TI - Questions, questions. PMID- 11863149 TI - New technology could reduce need for X-rays. PMID- 11863150 TI - Decontamination survey results published. PMID- 11863151 TI - The provision of general anaesthesia in dental practice, an end which had to come? AB - 31 December 2001 was the final day on which a general anaesthetic could be given in a dental practice in UK. Henceforth all dental treatment requiring a general anaesthetic will have to take place in a hospital setting, which has immediate access to critical care facilities. This will mark the end of the association between dental practice and general anaesthesia which dates back to the very first recorded clinical procedure performed under general anaesthesia, when in 1844, Horace Wells an American dentist, had a tooth removed by his assistant using nitrous oxide in Hartford, Connecticut, USA. PMID- 11863152 TI - Oral ulceration: a new and unusual complication. AB - Oral ulceration is a common oral mucosal disorder arising from a range of aetiologies but, apart from being associated with discomfort or pain, rarely results in complications apart from occasional scarring. This paper reports two patients with a history of minor aphthae who developed ulceration with increasingly severe pain, related to the onset of osteitis, and who then developed sequestra. PMID- 11863153 TI - A survey of antibiotic prescribing by maxillofacial consultants for dental extractions following radiotherapy to the oral cavity. AB - OBJECTIVES: To analyse the antibiotic prescribing trends for exodontia and minor oral surgery in patients with a history of radiotherapy. Also, to evaluate the use of hyperbaric oxygen as a separate modality in the prevention of osteoradionecrosis. DESIGN: A survey of antibiotic prescribing involving the analysis of a questionnaire which included the management of three patients with a history of head and neck malignancy. METHOD: The heads of the departments in each unit in the UK were sent a closed-response questionnaire. Antibiotic prescribing was assessed in three case scenarios. Case one referred to a patient that had surgery alone as part of the management of head and neck cancer, who needed the surgical removal of second molar roots. Case two referred to a patient that had surgery and adjuvant radiotherapy as part of the management of head and neck cancer, and needed the same procedure as in case one. Case three included a patient with a history of surgery and adjuvant radiotherapy, who required the extraction of three mobile and periodontally-involved lower incisors. Consultants were also asked about the use of hyperbaric oxygen in the patients who had radiotherapy as part of their treatment strategy. RESULTS: A total of 109 questionnaires were sent to all the Maxillofacial Units in the UK. The response rate was 73%. In patients with a history of radical surgery alone for the management of head and neck cancer, 20% of the surgeons advocated pre-operative antibiotics for the surgical removal of lower posterior teeth. In contrast, in the patient with a history of adjuvant radiotherapy 86% supported pre-operative antimicrobial use for the surgical removal of the same teeth. In the extraction of mobile and periodontally involved lower incisors in the previously irradiated patient, 63% of the clinicians supported pre-extraction antibiotics. Postoperative antibiotics were advocated in 52% in the first case, 89% in the second case and 71% in the third case. 34% of the clinicians advocated hyperbaric oxygen for surgical removal of posterior teeth and 15% for the extraction of mobile anterior teeth. CONCLUSIONS: The use of peri-extraction antibiotics was favoured among oral and maxillofacial surgeons. There was little enthusiasm towards the use of hyperbaric oxygen as a prophylactic measure, alone or in conjunction with an antimicrobial regime. Control randomised trials that will measure the effectiveness of hyperbaric oxygen and the appropriate use of antibiotics for prophylaxis in pre- and post-operative regimes are necessary to evaluate the use of these modalities. PMID- 11863154 TI - Effects of dental local anaesthetics in cardiac transplant recipients. AB - OBJECTIVE: To investigate the cardiovascular responses of cardiac transplant recipients to dental local anaesthetic solutions with and without epinephrine (adrenaline). MATERIALS AND METHODS: A clinical study employing 30 patients (20 cardiac transplant recipients and ten healthy) awaiting gingival or minor oral surgery under local anaesthesia receiving either 4.4 ml lidocaine (lignocaine) with 1:80,000 epinephrine or 4.4 ml 3% prilocaine with 0.031 U/ml felypressin. RESULTS: Cardiac transplant patients experienced a significant tachycardia 10 minutes after injection of the epinephrine-containing solution. No significant change in heart rate was detected after the injection of an epinephrine-free solution. Blood pressure was not affected. Periodontal surgery did not affect the responses to the local anaesthetics in the transplant recipients. CONCLUSIONS: The cardiovascular response to dental local anaesthesia in cardiac transplant recipients is governed by the solution injected. PMID- 11863155 TI - The effect of increasing vertical dimension of occlusion on facial aesthetics. AB - AIM: To investigate the effect of increasing the vertical dimension of occlusion on facial aesthetics. SETTING: General practice. METHOD: Questionnaires were sent to 96 patients who had been treated in the practice during the period of July 1998 to December 2000, resulting in an overall 72% response rate. All these patients had had their occlusal vertical dimension increased. Photographs of patients were taken before, during and after treatment. The questionnaire asked their opinion on the effects of the treatment on their facial features. To obtain an objective view to substantiate the opinions of the patients, a panel of five judges reviewed the before and after photographs and filled in their own questionnaires. RESULTS: Of the patients who responded to the questionnaire, 79.7% said they looked younger after the treatment. The panel thought 81.2% of the patients treated whose photographs they reviewed looked younger. CONCLUSION: Increasing the vertical dimension of occlusion can have far reaching effects on facial aesthetics, not just on the peri-oral areas but on the whole face. PMID- 11863156 TI - Child oral health promotion experiences in Northern Ireland. AB - It is a curious state of affairs that the children in Northern Ireland (NI) compared with their counterparts in the Republic of Ireland and Great Britain have some of the worst dental health. PMID- 11863157 TI - Effects of a limited class of nonlinearities on estimates of relative weights. AB - Perturbation analyses have been applied in recent years to determine the relative contribution of individual stimulus components in detection and discrimination tasks. Responses to stimulus samples are compared to stimulus parameters to determine the details of the decision rule. Often, a linear model is assumed and it is of interest to determine the relative contribution of different stimulus elements to the decision. Here, biases in estimated relative weights are considered for the case where the decision variable is given by D = (sigma(alphaiXn(i))k)m and the stimulus components, the Xi, are normally distributed, of equal variance, and mutually independent. The alphai are the "true" combination weights, and n, k, and m are positive reals. The method used to estimate relative weights is the correlation coefficient between the Xi and the observer's responses. Estimates of relative alphai do not depend on m but may depend on the mean values of the Xi and the values of n and k (a dependence on the variance, sigmai2, holds even for linear transformations). PMID- 11863158 TI - Modeling the influence of inherent envelope fluctuations in simultaneous masking experiments. AB - Masked thresholds are measured and simulated for bandpass-noise signals ranging in bandwidth from 4 to 256 Hz in the presence of a masking bandpass noise also ranging in bandwidth from 4 to 256 Hz. Signal and masker are centered at 2 kHz. To investigate the role of temporal processing in simultaneous masking, simulations were performed with the modulation-filterbank model by Dau et al. [J. Acoust. Soc Am. 102, 2906-2919 (1997)]. For a fixed masker bandwidth, thresholds are independent of the signal bandwidth as long as the signal bandwidth does not exceed the masker bandwidth and thresholds decrease with increasing masker bandwidth in those conditions. A simple modulation-low-pass filter (energy integrator) would be sufficient to describe the experimental results in those conditions. In contrast, the processing by a modulation filterbank is necessary to account for the conditions of "asymmetry of masking," where thresholds for signals with bandwidths larger than the masker bandwidth are much lower than those for the reversed condition. In those conditions, the modulation-filterbank model is able to use the inherent higher modulation frequencies of the signal as an additional cue. PMID- 11863159 TI - Discrimination of sound source velocity in human listeners. AB - The ability of six human subjects to discriminate the velocity of moving sound sources was examined using broadband stimuli presented in virtual auditory space. Subjects were presented with two successive stimuli moving in the frontal horizontal plane level with the ears, and were required to judge which moved the fastest. Discrimination thresholds were calculated for reference velocities of 15, 30, and 60 degrees/s under three stimulus conditions. In one condition, stimuli were centered on 0 degrees azimuth and their duration varied randomly to prevent subjects from using displacement as an indicator of velocity. Performance varied between subjects giving median thresholds of 5.5, 9.1, and 14.8 degrees/s for the three reference velocities, respectively. In a second condition, pairs of stimuli were presented for a constant duration and subjects would have been able to use displacement to assist their judgment as faster stimuli traveled further. It was found that thresholds decreased significantly for all velocities (3.8, 7.1, and 9.8 degrees/s), suggesting that the subjects were using the additional displacement cue. The third condition differed from the second in that the stimuli were "anchored" on the same starting location rather than centered on the midline, thus doubling the spatial offset between stimulus endpoints. Subjects showed the lowest thresholds in this condition (2.9, 4.0, and 7.0 degrees/s). The results suggested that the auditory system is sensitive to velocity per se, but velocity comparisons are greatly aided if displacement cues are present. PMID- 11863160 TI - Frequency-to-electrode allocation and speech perception with cochlear implants. AB - The hypothesis was investigated that selectively increasing the discrimination of low-frequency information (below 2600 Hz) by altering the frequency-to-electrode allocation would improve speech perception by cochlear implantees. Two experimental conditions were compared, both utilizing ten electrode positions selected based on maximal discrimination. A fixed frequency range (200-10513 Hz) was allocated either relatively evenly across the ten electrodes, or so that nine of the ten positions were allocated to the frequencies up to 2600 Hz. Two additional conditions utilizing all available electrode positions (15-18 electrodes) were assessed: one with each subject's usual frequency-to-electrode allocation; and the other using the same analysis filters as the other experimental conditions. Seven users of the Nucleus CI22 implant wore processors mapped with each experimental condition for 2-week periods away from the laboratory, followed by assessment of perception of words in quiet and sentences in noise. Performance with both ten-electrode maps was significantly poorer than with both full-electrode maps on at least one measure. Performance with the map allocating nine out of ten electrodes to low frequencies was equivalent to that with the full-electrode maps for vowel perception and sentences in noise, but was worse for consonant perception. Performance with the evenly allocated ten electrode map was equivalent to that with the full-electrode maps for consonant perception, but worse for vowel perception and sentences in noise. Comparison of the two full-electrode maps showed that subjects could fully adapt to frequency shifts up to ratio changes of 1.3, given 2 weeks' experience. Future research is needed to investigate whether speech perception may be improved by the manipulation of frequency-to-electrode allocation in maps which have a full complement of electrodes in Nucleus implants. PMID- 11863161 TI - On the influence of laryngeal pathologies on acoustic and electroglottographic jitter measures. AB - This study compared acoustic and electroglottographic (EGG) jitter from [a] vowels of 103 dysphonic speakers. The EGG recordings were chosen according to their intensity, signal-to-noise ratio, and percentage of unvoiced intervals, while acoustic signals were selected based on voicing detection and the reliability of jitter extraction. The agreement between jitter measures was expressed numerically as a normalized difference. In 63.1% (65/103) of the cases the differences fell within +/-22.5%. Positive differences above +22.5% were associated with increased acoustic jitter and occurred in 12.6% (13/103) of the speakers. These were, typically, cases of small nodular lesions without problems in the posterior larynx. On the other hand, substantial rises in EGG jitter leading to differences below -22.5% took place in 24.3% (25/103) of the speakers and were related to hyperfunctional voices, creaky-like voices, small laryngeal asymmetries affecting the arytenoids, or small-to-moderate glottal chinks. A clinically relevant outcome of the study was the possibility of detecting gentle laryngeal asymmetries among cases of large unilateral increase in EGG jitter. These asymmetries can be linked with vocal problems that are often overlooked in endoscopic examinations. PMID- 11863162 TI - Synchrony capture hypothesis fails to account for effects of amplitude on voicing perception. AB - Kluender et al. [J. Acoust. Soc. Am. 97, 2552-2567 (1995)] reported that overall stimulus amplitude affects perception of the voicing contrast in syllable-initial stops as a function of frequency separation between the first formant (F1) and higher formants (F2, F3). These results were offered as support for a hypothesis that [- voice] could be signaled by a shift in the temporal pattern of neural firing from synchronizing to energy at frequencies of F2 and F3 to synchronizing to energy near F1. Several predictions from this "synchrony capture hypothesis" were tested in the current study. In all cases the hypothesis was not supported. Effect of stimulus amplitude (increased voiceless responses with higher amplitude) was maintained when there was no cutback in F1 or when F2 and F1 energy bands were presented dichotically. In further tests of the hypothesis, voice-voiceless series were created that maintained periodic energy throughout the syllable (with F1 cutback signaling voicing). Energy just below the frequency of F2 and energy above F1 were presented dichotically. Thus, at the periphery there was no competition between frequencies near F2 and lower frequencies. In contrast to predictions of the "synchrony capture hypothesis," overall amplitude still had an effect on voice-voiceless identifications. PMID- 11863163 TI - Detecting stop consonants in continuous speech. AB - The problem of implementing a detector for stop consonants in continuously spoken speech is considered. The problem is posed as one of finding an optimal filter (linear or nonlinear) that operates on a particular appropriately chosen representation, and ideally outputs a 1 when a stop occurs and 0 otherwise. The performance of several variants of a canonical stop detector is discussed and its implications for human and machine speech recognition is considered. PMID- 11863164 TI - On application of adaptive decorrelation filtering to assistive listening. AB - This paper describes an application of the multichannel signal processing technique of adaptive decorrelation filtering to the design of an assistive listening system. A simulated "dinner table" scenario was studied. The speech signal of a desired talker was corrupted by three simultaneous speech jammers and by a speech-shaped diffusive noise. The technique of adaptive decorrelation filtering processing was used to extract the desired speech from the interference speech and noise. The effectiveness of the assistive listening system was evaluated by observing improvements in A-weighted signal-to-noise ratio (SNR) and in sentence intelligibility, where the latter was evaluated in a listening test with eight normal hearing subjects and three subjects with hearing impairments. Significant improvements in SNR and sentence intelligibility were achieved with the use of the assistive listening system. For subjects with normal hearing, the speech reception threshold was improved by 3 to 5 dBA, and for subjects with hearing impairments, the threshold was improved by 4 to 8 dBA. PMID- 11863165 TI - An overlapping-feature-based phonological model incorporating linguistic constraints: applications to speech recognition. AB - Modeling phonological units of speech is a critical issue in speech recognition. In this paper, our recent development of an overlapping-feature-based phonological model that represents long-span contextual dependency in speech acoustics is reported. In this model, high-level linguistic constraints are incorporated in automatic construction of the patterns of feature-overlapping and of the hidden Markov model (HMM) states induced by such patterns. The main linguistic information explored includes word and phrase boundaries, morpheme, syllable, syllable constituent categories, and word stress. A consistent computational framework developed for the construction of the feature-based model and the major components of the model are described. Experimental results on the use of the overlapping-feature model in an HMM-based system for speech recognition show improvements over the conventional triphone-based phonological model. PMID- 11863166 TI - Ultrasound-induced lung hemorrhage: role of acoustic boundary conditions at the pleural surface. AB - In a previous study [J. Acoust. Soc. Am. 108, 1290 (2000)] the acoustic impedance difference between intercostal tissue and lung was evaluated as a possible explanation for the enhanced lung damage with increased hydrostatic pressure, but the hydrostatic-pressure-dependent impedance difference alone could not explain the enhanced occurrence of hemorrhage. In that study, it was hypothesized that the animal's breathing pattern might be altered as a function of hydrostatic pressure, which in turn might affect the volume of air inspired and expired. The acoustic impedance difference between intercostal tissue and lung would be affected with altered lung inflation, thus altering the acoustic boundary conditions. In this study, 12 rats were exposed to 3 volumes of lung inflation (inflated: approximately tidal volume; half-deflated: half-tidal volume; deflated: lung volume at functional residual capacity), 6 rats at 8.6-MPa in situ peak rarefactional pressure (MI of 3.1) and 6 rats at 16-MPa in situ peak rarefactional pressure (MI of 5.8). Respiration was chemically inhibited and a ventilator was used to control lung volume and respiratory frequency. Superthreshold ultrasound exposures of the lungs were used (3.1-MHz, 1000-Hz PRF, 1.3-micros pulse duration, 10-s exposure duration) to produce lesions. Deflated lungs were more easily damaged than half-deflated lungs, and half-deflated lungs were more easily damaged than inflated lungs. In fact, there were no lesions observed in inflated lungs in any of the rats. The acoustic impedance difference between intercostal tissue and lung is much less for the deflated lung condition, suggesting that the extent of lung damage is related to the amount of acoustic energy that is propagated across the pleural surface boundary. PMID- 11863167 TI - Quantitative investigation of acoustic streaming in blood. AB - Acoustic streaming may have practical utility in diagnostic medical ultrasound in distinguishing between stagnant blood and tissue as well as clotted and unclotted blood. This distinction can be difficult with conventional ultrasound but have high value in managing trauma patients with internal hemorrhage. Ultrasound energy applies a force to blood by momentum transfer, resulting in bulk streaming that is a function of the acoustic attenuation, sound speed, acoustic intensity, blood viscosity, and the boundary conditions posed by the geometry around the hematoma. A simple tubular model was studied analytically, by finite element simulation, and experimentally by in vitro measurement. The simulation agreed closely with measurements while the analytic solutions were found to be valid only for beam diameters approximating the diameter of the tubular channel. Experimentally, the acoustic streaming in blood decreased as the blood began to clot and the streaming flow was not detected in clotted blood. In contrast, the echogenicity of the same blood samples did not change appreciably from the unclotted to the clotted state for the stagnant blood studied. Streaming detection appears to offer a potential tool for improving hemorrhage diagnosis. PMID- 11863168 TI - Relative loudness of low- and high-frequency bands of speech-shaped babble, including the influence of bandwidth and input level (L). PMID- 11863172 TI - The role of Biot slow waves in electroseismic wave phenomena. AB - The electromagnetic fields that are generated as a spherical seismic wave (either P or S) traverses an interface separating two porous materials are numerically modeled both with and without the generation of Biot slow waves at the interface. In the case of an incident fast-P wave, the predicted electric-field amplitudes when slow waves are neglected can easily be off by as much as an order of magnitude. In the case of an incident S wave, the error is much smaller (typically on the order of 10% or less) because not much S-wave energy gets converted into slow waves. In neglecting the slow waves, only six plane waves (reflected and transmitted fast-P, S, and EM waves) are available with which to match the eight continuity conditions that hold at each interface. This overdetermined problem is solved by placing weights on the eight continuity conditions so that those conditions that are most important for obtaining the proper response are emphasized. It is demonstrated that when slow waves are neglected, it is best to also neglect the continuity of the Darcy flow and fluid pressure across an interface. The principal conclusion of this work is that to properly model the electromagnetic (EM) fields generated at an interface by an incident seismic wave, the full Biot theory that allows for generation of slow waves must be employed. PMID- 11863173 TI - Measurement of the B/A nonlinearity parameter under high pressure: application to water. AB - An experimental apparatus was developed to measure, over a wide range of pressure, the acoustical nonlinear parameter B/A with an uncertainty of 2.2% in order to study the influence of pressure on the value of this parameter in liquids. The experimental technique rests on an improved thermodynamic method which uses a highly sensitive phase comparison technique to measure the change in speed of sound with pressure. The apparatus was then used to measure B/A in water within the pressure range from 0.1 to 50 MPa and at temperatures of between 303.15 and 373.15 K. The data obtained were compared with those in the literature which come from numerical derivation of speed of sound measurements. PMID- 11863174 TI - Refracted arrival waves in a zone of silence from a finite thickness mixing layer. AB - Refracted arrival waves which propagate in the zone of silence of a finite thickness mixing layer are analyzed using geometrical acoustics in two dimensions. Here, two simplifying assumptions are made: (i) the mean flow field is transversely sheared, and (ii) the mean velocity and temperature profiles approach the free-stream conditions exponentially. Under these assumptions, ray trajectories are analytically solved, and a formula for acoustic pressure amplitude in the far field is derived in the high-frequency limit. This formula is compared with the existing theory based on a vortex sheet corresponding to the low-frequency limit. The analysis covers the dependence on the Mach number as well as on the temperature ratio. The results show that both limits have some qualitative similarities, but the amplitude in the zone of silence at high frequencies is proportional to omega(-1/2), while that at low frequencies is proportional to omega(-3/2), omega being the angular frequency of the source. PMID- 11863175 TI - The connection between sound production and jet structure of the supersonic impinging jet. AB - An experimental investigation into the sound-producing characteristics of moderately and highly underexpanded supersonic impinging jets exhausting from a round convergent nozzle is presented. The production of large plate tones by impingement on a square plate with a side dimension equal to 12 nozzle exit diameters is studied using random and phase-locked shadowgraph photography. Discrete frequency sound is produced in the near-wall region of the jet when a Mach disk occurs upstream of the standoff shock wave. Tones cease when the plate distance is approximately 2.2 free-jet cell lengths and the first and second shock waves are located in the free-jet positions. The production of impulsive sound appears to be associated with the collapse of the standoff shock wave during a portion of the oscillation cycle. Results from unsteady plate-pressure measurements indicate that plane-wave motion occurs in the impingement region and a secondary pressure maximum is observed on the plate adjacent to the flow region where sound appears to originate. PMID- 11863176 TI - Extraction of acoustic normal mode depth functions using vertical line array data. AB - A method for extracting the normal modes of acoustic propagation in the shallow ocean from sound recorded on a vertical line array (VLA) of hydrophones as a source travels nearby is presented. The mode extraction is accomplished by performing a singular value decomposition (SVD) of individual frequency components of the signal's temporally averaged, spatial cross-spectral density matrix. The SVD produces a matrix containing a mutually orthogonal set of basis functions, which are proportional to the depth-dependent normal modes, and a diagonal matrix containing the singular values, which are proportional to the modal source excitations and mode eigenvalues. The conditions under which the method is expected to work are found to be (1) sufficient depth sampling of the propagating modes by the VLA receivers; (2) sufficient source-VLA range sampling, and (3) sufficient range interval traversed by the source. The mode extraction method is applied to data from the Area Characterization Test II, conducted in September 1993 in the Hudson Canyon Area off the New Jersey coast. Modes are successfully extracted from cw tones recorded while (1) the source traveled along a range-independent track with constant bathymetry and (2) the source traveled up slope with gradual changes in bathymetry. In addition, modes are successfully extracted at multiple frequencies from ambient noise. PMID- 11863177 TI - Study of a novel range-dependent propagation effect with application to the axial injection of signals from the Kaneohe source. AB - A novel range-dependent propagation effect occurs when a source is placed on the seafloor in shallow water with a downward refracting sound speed profile, and sound waves propagate down a slope into deep water. Under these conditions, small grazing-angle sound waves slide along the bottom downward and outward from the source until they reach the depth of the sound channel axis in deep water, where they are detached from the sloping bottom and continue to propagate outward near the sound channel axis. This "mudslide" effect is one of a few robust and predictable acoustic propagation effects that occur in range-dependent ocean environments. As a consequence of this effect, a bottom mounted source in shallow water can inject a significant amount of acoustic energy into the axis of the deep ocean sound channel that can then propagate to very long ranges. Numerical simulations with a full-wave range-dependent acoustic model show that the Kaneohe experiment had the appropriate source, bathymetry, and sound speed profiles that allows this effect to operate efficiently. This supports the interpretation that some of the near-axial acoustic signals, received near the coast of California from the bottom mounted source located in shallow water in Kaneohe Bay, Oahu, Hawaii, were injected into the sound channel of the deep Pacific Ocean by this mechanism. Numerical simulations suggest that the mudslide effect is robust. PMID- 11863179 TI - Acoustic propagation through anisotropic internal wave fields: transmission loss, cross-range coherence, and horizontal refraction. AB - Results of a computer simulation study are presented for acoustic propagation in a shallow water, anisotropic ocean environment. The water column is characterized by random volume fluctuations in the sound speed field that are induced by internal gravity waves, and this variability is superimposed on a dominant summer thermocline. Both the internal wave field and resulting sound speed perturbations are represented in three-dimensional (3D) space and evolve in time. The isopycnal displacements consist of two components: a spatially diffuse, horizontally isotropic component and a spatially localized contribution from an undular bore (i.e., a solitary wave packet or solibore) that exhibits horizontal (azimuthal) anisotropy. An acoustic field is propagated through this waveguide using a 3D parabolic equation code based on differential operators representing wide-angle coverage in elevation and narrow-angle coverage in azimuth. Transmission loss is evaluated both for fixed time snapshots of the environment and as a function of time over an ordered set of snapshots which represent the time-evolving sound speed distribution. Horizontal acoustic coherence, also known as transverse or cross-range coherence, is estimated for horizontally separated points in the direction normal to the source-receiver orientation. Both transmission loss and spatial coherence are computed at acoustic frequencies 200 and 400 Hz for ranges extending to 10 km, a cross-range of 1 km, and a water depth of 68 m. Azimuthal filtering of the propagated field occurs for this environment, with the strongest variations appearing when propagation is parallel to the solitary wave depressions of the thermocline. A large anisotropic degradation in horizontal coherence occurs under the same conditions. Horizontal refraction of the acoustic wave front is responsible for the degradation, as demonstrated by an energy gradient analysis of in-plane and out-of-plane energy transfer. The solitary wave packet is interpreted as a nonstationary oceanographic waveguide within the water column, preferentially funneling acoustic energy between the thermocline depressions. PMID- 11863180 TI - Velocity dispersion in water-saturated granular sediment. AB - Recent experiments in the Gulf of Mexico have yielded a wealth of information on the environmental conditions and geoacoustic response of a uniform sand stratum immediately beneath the seafloor. A comparison of p-wave velocities measured at low (125 Hz) and high (11-50 kHz) frequencies in this layer indicates that there is a significant amount of velocity dispersion that occurs in the interval between these extremes. This narrow-band dispersion, which is not consistent with the often-used assumption of a nearly constant-Q model, is in accordance with the predictions of the Biot theory. It results from viscous damping in the fluid as it moves relative to the skeletal frame. Other recent field data that support this conclusion are presented. PMID- 11863181 TI - Seabed classification from acoustic profiling data using the similarity index. AB - We introduce the similarity index (SI) for the classification of the sea floor from acoustic profiling data. The essential part of our approach is the singular value decomposition of the data to extract a signal coherent trace-to-trace using the Karhunen-Loeve transform. SI is defined as the percentage of the energy of the coherent part contained in the bottom return signals. Important aspects of SI are that it is easily computed and that it represents the textural roughness of the sea floor as a function of grain size, hardness, and a degree of sediment sorting. In a real data example, we classified a section of the sea floor off Cheju Island south of the Korean Peninsula and compared the result with the sedimentology defined from direct sediment sampling and side scan sonar records. The comparison shows that SI can efficiently discriminate the bottom properties by delineating sediment-type boundaries and transition zones in more detail. Therefore, we propose that SI is an effective parameter for geoacoustic modeling. PMID- 11863183 TI - Modeling of high-frequency propagation in inhomogeneous background random media. AB - When a high-frequency electromagnetic wave propagates in a complicated scattering environment, the contribution at the observer is usually composed of a number of field species arriving along different ray trajectories. In order to describe each contribution separately the parabolic extension along an isolated ray trajectory in an inhomogeneous background medium was performed. This leads to the parabolic wave equation along a deterministic ray trajectory in a randomly perturbed medium with the possibility of presenting the solution of the high frequency field and the higher-order coherence functions in the functional path integral form. It is shown that uncertainty considerations play an important role in relating the path-integral solutions to the approximate asymptotic solutions. The solutions for the high-frequency propagators derived in this work preserve the random information accumulated along the propagation path and therefore can be applied to the analysis of double-passage effects where the correlation between the forward-backward propagating fields has to be accounted for. This results in double-passage algorithms, which have been applied to analyze the resolution of two point scatterers. Under strong scattering conditions, the backscattering effects cannot be neglected and the ray trajectories cannot be treated separately. The final part is devoted to the generalized parabolic extension method applied to the scalar Helmholtz's equation, and possible approximations for obtaining numerically manageable solutions in the presence of random media. PMID- 11863182 TI - A phase regulated back wave propagation technique for geoacoustic inversion. AB - An inversion method based on the concept of back wave propagation (BWP) is described in this paper for estimation of geoacoustic parameters from acoustic field data. A phase-regulation technique is introduced to increase the sensitivity of the method for geoacoustic model parameters having low sensitivity. The case of data consisting of signal plus additive noise is also addressed. It is shown theoretically that the sensitivity can be increased by a factor alpha using the phase regulation procedure, and that the spatial resolution of signal energy that is concentrated by BWP at the known source position is increased when a increases. This result suggests an effective criterion for use in the inversion, based on the spatial distribution of signal energy around the true source location. The basis for the criterion is the spatial variance of the back-propagated pressure field in a window around the known source location. A multistep search process is proposed to avoid using a complicated multidimensional search process. Inversion results from both simulations and experimental data are given. The real data were taken from the Pacific Shelf experiment carried out in shallow water off the West Coast of Vancouver Island in the Northeast Pacific Ocean. PMID- 11863184 TI - Broadband time-reversing array retrofocusing in noisy environments. AB - Acoustic time reversal is a promising technique for spatial and temporal focusing of sound in unknown environments. Acoustic time reversal can be implemented with an array of transducers that listens to a remote sound source and then transmits a time-reversed version of what was heard. In a noisy environment, the performance of such a time-reversing array (TRA) will be degraded because the array will receive and transmit noise, and the intended signal may be masked by ambient noise at the retrofocus location. This article presents formal results for the signal-to-noise ratio at the intended focus (SNRf) for TRAs that receive and send finite-duration broadband signals in noisy environments. When the noise is homogeneous and uncorrelated, and a broadcast power limitation sets the TRA's electronic amplification, the formal results can be simplified to an algebraic formula that includes the characteristics of the signal, the remote source, the TRA, and the noisy environment. Here, SNRf is found to be proportional to the product of the signal bandwidth and the duration of the signal pulse after propagation through the environment. Using parabolic-equation propagation simulations, the formal results for SNRf are illustrated for a shallow water environment at source-array ranges of 1 to 40 km and bandwidths from several tens of Hz to more than 500 Hz for a signal center frequency of 500 Hz. Shallow-water TRA noise rejection is predicted to be superior to that possible in free space because TRAs successfully exploit multipath-propagation. PMID- 11863185 TI - Numerical investigations of flow and energy fields near a thermoacoustic couple. AB - The flow field and the energy transport near thermoacoustic couples are simulated using a 2D full Navier-Stokes solver. The thermoacoustic couple plate is maintained at a constant temperature; plate lengths, which are "short" and "long" compared with the particle displacement lengths of the acoustic standing waves, are tested. Also investigated are the effects of plate spacing and the amplitude of the standing wave. Results are examined in the form of energy vectors, particle paths, and overall entropy generation rates. These show that a net heat pumping effect appears only near the edges of thermoacoustic couple plates, within about a particle displacement distance from the ends. A heat-pumping effect can be seen even on the shortest plates tested when the plate spacing exceeds the thermal penetration depth. It is observed that energy dissipation near the plate increases quadratically as the plate spacing is reduced. The results also indicate that there may be a larger scale vortical motion outside the plates which disappears as the plate spacing is reduced. PMID- 11863186 TI - A model and experimental study of fiber orientation effects on shear wave propagation through composite laminates. AB - The strong elastic anisotropy of the discrete unidirectional plies in a composite laminate interacts sensitively with the polarization direction of a shear ultrasonic wave propagating in the thickness direction. The transmitted shear wave can therefore be used to detect errors in the ply orientation and stacking sequence of a laminate. The sensitivity is particularly high when the polarization directions of the shear wave transmitter and receiver are orthogonal to each other. To understand the interaction between normal-incident shear waves and ply orientations in a laminate, a complete analytical model was developed using local and global transfer matrices. The model predicted the transmitted signal amplitude as a function of polarization angle of the transmitter and time (or frequency) for a given laminate and input signal. To alleviate the experimental problems associated with shear wave coupling, electromagnetic acoustic transducers (EMATs) and metal delay lines were used in the angular scan of the transmitted signal. The EMAT system had the added advantage of being applicable to uncured composite laminates. Experiments were performed on both cured and uncured laminates with common layups for model verification. The sensitivity of the measured shear wave signals to fiber misorientation and stacking sequence errors was also demonstrated. PMID- 11863188 TI - Noise in miniature microphones. AB - The internal noise spectrum in miniature electret microphones of the type used in the manufacture of hearing aids is measured. An analogous circuit model of the microphone is empirically fit to the measured data and used to determine the important sources of noise within the microphone. The dominant noise source is found to depend on the frequency. Below 40 Hz and above 9 kHz, the dominant source is electrical noise from the amplifier circuit needed to buffer the electrical signal from the microphone diaphragm. Between approximately 40 Hz and 1 kHz, the dominant source is thermal noise originating in the acoustic flow resistance of the small hole pierced in the diaphragm to equalize barometric pressure. Between approximately 1 kHz and 9 kHz, the noise originates in the acoustic flow resistances of sound entering the microphone and propagating to the diaphragm. To further reduce the microphone internal noise in the audio band requires attacking these sources. A prototype microphone having reduced acoustical noise is measured and discussed. PMID- 11863189 TI - Acoustic streaming induced by ultrasonic flexural vibrations and associated enhancement of convective heat transfer. AB - Acoustic streaming induced by ultrasonic flexural vibrations and the associated convection enhancement are investigated. Acoustic streaming pattern, streaming velocity, and associated heat transfer characteristics are experimentally observed. Moreover, analytical analysis based on Nyborg's formulation is performed along with computational fluid dynamics (CFD) simulation using a numerical solver CFX 4.3. Two distinctive acoustic streaming patterns in half wavelength of the flexural vibrations are observed, which agree well with the theory. However, acoustic streaming velocities obtained from CFD simulation, based on the incompressible flow assumption, exceed the theoretically estimated velocity by a factor ranging from 10 to 100, depending upon the location along the beam. Both CFD simulation and analytical analysis reveal that the acoustic streaming velocity is proportional to the square of the vibration amplitude and the wavelength of the vibrating beam that decreases with the excitation frequency. It is observed that the streaming velocity decreases with the excitation frequency. Also, with an open-ended channel, a substantial increase in streaming velocity is observed from CFD simulations. Using acoustic streaming, a temperature drop of 40 degrees C with a vibration amplitude of 25 microm at 28.4 kHz is experimentally achieved. PMID- 11863190 TI - Grazing instabilities and post-bifurcation behavior in an impacting string. AB - A theoretical and experimental investigation of the nonlinear dynamic response of a periodically excited string subject to a knife-edge amplitude restraint is presented. The amplitude restraint creates an impact condition as the amplitude of the response grows. The focus of this work is on the influence of a grazing instability; this zero-velocity impact event leads to complicated, post bifurcation behavior ranging from multifrequency, periodic motion to chaos. In addition to looking at the response numerically, parameter combinations leading to an incidence of grazing are clearly identified in the excitation force excitation frequency parameter space using a multiple scales perturbation analysis. Modeling issues, numerical difficulties, and experimental limitations are also discussed. PMID- 11863191 TI - Scattering and active acoustic control from a submerged spherical shell. AB - This paper is concerned with the scattering from a submerged (heavy fluid) bilaminate spherical shell composed of an outer layer of steel, and an inner layer of radially polarized piezoelectric material. The methodology used includes separation formulas for the stresses and displacements, which in turn are used (coupled with spherical harmonics) to reduce the governing equations to linear systems of ordinary differential equations. This technique uses the full equations of elasticity rather than any of the various thin-shell approximations in determining the axisymmetric scattering from a shell, normal modes of vibration for the shell, as well as voltages necessary for annihilation of a scattered pressure due to insonification of the shell by an incident plane wave. PMID- 11863192 TI - Active vibroacoustic control with multiple local feedback loops. AB - When multiple actuators and sensors are used to control the vibration of a panel, or its sound radiation, they are usually positioned so that they couple into specific modes and are all connected together with a centralized control system. This paper investigates the physical effects of having a regular array of actuator and sensor pairs that are connected only by local feedback loops. An array of 4 x 4 force actuators and velocity sensors is first simulated, for which such a decentralized controller can be shown to be unconditionally stable. Significant reductions in both the kinetic energy of the panel and in its radiated sound power can be obtained for an optimal value of feedback gain, although higher values of feedback gain can induce extra resonances in the system and degrade the performance. A more practical transducer pair, consisting of a piezoelectric actuator and velocity sensor, is also investigated and the simulations suggest that a decentralized controller with this arrangement is also stable over a wide range of feedback gains. The resulting reductions in kinetic energy and sound power are not as great as with the force actuators, due to the extra resonances being more prominent and at lower frequencies, but are still worthwhile. This suggests that an array of independent modular systems, each of which included an actuator, a sensor, and a local feedback control loop, could be a simple and robust method of controlling broadband sound transmission when integrated into a panel. PMID- 11863193 TI - Reduction of electronic delay in active noise control systems--a multirate signal processing approach. AB - Electronic delay has been a critical problem in active noise control (ANC) systems. This is true whether a feedforward structure or a feedback structure is adopted. In particular, excessive delays would create a causality problem in a feedforward ANC system of a finite-length duct. This paper suggests a multirate signal-processing approach for minimizing the electronic delay in the control loop. In this approach, digital controllers are required in decimation and interpolation of discrete-time signals. The computation efficiency is further enhanced by a polyphase method, where the phases of low-pass finite impulse response (FIR) filters must be carefully designed to avoid unnecessary delays. Frequency domain optimization procedures based on H1, H2, and Hinfinity norms, respectively, are utilized in the FIR filter design. The proposed method was implemented by using a floating-point digital signal processor. Experimental results showed that the multirate approach remains effective for suppressing a broadband (200-600 Hz) noise in a duct with a minimum upstream measurement microphone placement of 20 cm. PMID- 11863195 TI - Effect of noise and occupancy on optimal reverberation times for speech intelligibility in classrooms. AB - The question of what is the optimal reverberation time for speech intelligibility in an occupied classroom has been studied recently in two different ways, with contradictory results. Experiments have been performed under various conditions of speech-signal to background-noise level difference and reverberation time, finding an optimal reverberation time of zero. Theoretical predictions of appropriate speech-intelligibility metrics, based on diffuse-field theory, found nonzero optimal reverberation times. These two contradictory results are explained by the different ways in which the two methods account for background noise, both of which are unrealistic. To obtain more realistic and accurate predictions, noise sources inside the classroom are considered. A more realistic treatment of noise is incorporated into diffuse-field theory by considering both speech and noise sources and the effects of reverberation on their steady-state levels. The model shows that the optimal reverberation time is zero when the speech source is closer to the listener than the noise source, and nonzero when the noise source is closer than the speech source. Diffuse-field theory is used to determine optimal reverberation times in unoccupied classrooms given optimal values for the occupied classroom. Resulting times can be as high as several seconds in large classrooms; in some cases, optimal values are unachievable, because the occupants contribute too much absorption. PMID- 11863196 TI - On the sampling conditions for reconstruction of an acoustic field from a finite sound source. AB - A simple and accurate method for the estimation of ultrasonic transducer fields is developed. In the method, the angular spectrum is employed to evaluate the three-dimensional propagation from a measured plane to an arbitrary parallel plane. The implementation uses a discrete convolution that is described in detail. Relative to conventional spatial-frequency representations, the implementation of the angular spectrum method in this paper has the advantage of being free from artifacts, enabling sample spacing to be greater than one half wavelength, using memory efficiently, and interpolating the measured data. The loosened sampling requirement and natural interpolation of the method permit efficient reconstruction of the full three-dimensional acoustic field from a coarse sound pressure measurement on single plane. PMID- 11863197 TI - Factors contributing to bone conduction: the middle ear. AB - Measurement of the motion of the malleus umbo and stapes footplate during bone conduction (BC) stimulation was conducted in vitro in 26 temporal bones using a laser Doppler vibrometer over the frequency range 0.1 to 10 kHz. For lower frequencies, both ossicular sites followed the motion of the temporal bone. The differential motion between the malleus and the surrounding bone was greater than the differential motion of the stapes footplate; both resonated near 1.5 kHz. Different lesions were shown to affect the response: (1) a mass attached to the umbo lowered the resonance frequency of the ossicular vibration; (2) fixation of either the malleus or stapes increased the stiffness and shifted the resonance frequency upward; and (3) dislocation of the incudo-stapedial joint did not significantly affect the ossicular vibration. The sound radiated from the tympanic membrane was approximately 85 dB SPL at an umbo differential velocity of 1 mm/s for low frequencies in an open ear canal and about 10 dB higher for an occluded one; at higher frequencies (above 2 kHz) resonances of the canal determine the response. It was also found that the motion between the footplate and promontory was within 5 dB when the specimen was stimulated orthogonal to the vibration direction of the ossicles than in line with the same. Measurement of the differential motion of the umbo in one live human skull gave similar response as the average result from the temporal bone specimens. PMID- 11863198 TI - Mapping ear canal movement using area-based surface matching. AB - Movement of the external ear canal, associated with jaw motion, relative to the concha region of the pinna has been studied. Pairs of open-jaw and closed-jaw impressions were taken of 14 ears from 10 subjects. Three-dimensional coordinate data were obtained from the concha and the anterior surface of the canal using a reflex microscope. Proprietary area-based matching software was used to evaluate distortion of the two surfaces between the two jaw positions. The canal data from each pair were placed into the same coordinate system with their respective concha regions aligned. Difference maps of the canal data were used to demonstrate the amount of anterior-posterior (A-P), superior-inferior (S-I), and medial-lateral (M-L) movement, relative to the concha, that occurred between the open- and closed-jaw impressions. The concha regions did not undergo significant deformation. The canal regions underwent varying amounts of deformation with all canals conforming within an rms of 136 microm across the entire surface. The majority of canals underwent significant movement relative to the concha. M-L movement ranged from +2.0 to -3.8 mm; eight canals moved laterally, five moved medially, and two showed no movement. S-I movement ranged from +3.7 to -2.7 mm; nine canals moved inferiorly, two moved superiorly, and three showed no movement. A-P movement ranged between +7.5 and -8.5 mm, with five canals moving anteriorly, three posteriorly, and four in a mixed fashion. This study has shown the variability of canal movement relative to the concha and does not support previous reports that suggest that the ear canal only widens with jaw opening. PMID- 11863199 TI - On the detection of early cochlear damage by otoacoustic emission analysis. AB - Theoretical considerations and experimental evidence suggest that otoacoustic emission parameters may be used to reveal early cochlear damage, even before it can be diagnosed by standard audiometric techniques. In this work, the statistical distributions of a set of otoacoustic emission parameters chosen as candidates for the early detection of cochlear damage (global and band reproducibility, response level, signal-to-noise ratio, spectral latency, and long-lasting otoacoustic emission presence) were analyzed in a population of 138 ears. These ears have been divided, according to a standard audiometric test, in three classes: (1) ears of nonexposed bilaterally normal subjects, (2) normal ears of subjects with unilateral noise-induced high-frequency hearing loss, and (3) their hearing impaired ears. For all analyzed parameters, a statistically significant difference was found between classes 1 and 2. This difference largely exceeds the difference observed between classes 2 and 3. This fact suggests that the noise exposure, which was responsible for the unilateral hearing loss, also caused subclinical damage in the contralateral, audiometrically normal, ear. This is a clear indication that otoacoustic emission techniques may be able to early detect subclinical damages. PMID- 11863200 TI - The mechanical waveform of the basilar membrane. IV. Tone and noise stimuli. AB - Analysis of mechanical cochlear responses to wide bands of random noise clarifies many effects of cochlear nonlinearity. The previous paper [de Boer and Nuttall, J. Acoust. Soc. Am. 107, 1497-1507 (2000)] illustrates how closely results of computations in a nonlinear cochlear model agree with responses from physiological experiments. In the present paper results for tone stimuli are reported. It was found that the measured frequency response for pure tones differs little from the frequency response associated with a noise signal. For strong stimuli, well into the nonlinear region, tones have to be presented at a specific level with respect to the noise for this to be true. In this report the nonlinear cochlear model originally developed for noise analysis was modified to accommodate pure tones. For this purpose the efficiency with which outer hair cells modify the basilar-membrane response was made into a function of cochlear location based on local excitation. For each experiment, the modified model is able to account for the experimental findings, within 1 or 2 dB. Therefore, the model explains why the type of filtering that tones undergo in the cochlea is essentially the same as that for noise signals (provided the tones are presented at the appropriate level). PMID- 11863201 TI - Neural activity associated with distinguishing concurrent auditory objects. AB - The neural processes underlying concurrent sound segregation were examined by using event-related brain potentials. Participants were presented with complex sounds comprised of multiple harmonics, one of which could be mistuned so that it was no longer an integer multiple of the fundamental. In separate blocks of trials, short-, middle-, and long-duration sounds were presented and participants indicated whether they heard one sound (i.e., buzz) or two sounds (i.e., buzz plus another sound with a pure-tone quality). The auditory stimuli were also presented while participants watched a silent movie in order to evaluate the extent to which the mistuned harmonic could be automatically detected. The perception of the mistuned harmonic as a separate sound was associated with a biphasic negative-positive potential that peaked at about 150 and 350 ms after sound onset, respectively. Long duration sounds also elicited a sustained potential that was greater in amplitude when the mistuned harmonic was perceptually segregated from the complex sound. The early negative wave, referred to as the object-related negativity (ORN), was present during both active and passive listening, whereas the positive wave and the mistuning-related changes in sustained potentials were present only when participants attended to the stimuli. These results are consistent with a two-stage model of auditory scene analysis in which the acoustic wave is automatically decomposed into perceptual groups that can be identified by higher executive functions. The ORN and the positive waves were little affected by sound duration, indicating that concurrent sound segregation depends on transient neural responses elicited by the discrepancy between the mistuned harmonic and the harmonic frequency expected based on the fundamental frequency of the incoming stimulus. PMID- 11863202 TI - Quantifying the implications of nonlinear cochlear tuning for auditory-filter estimates. AB - The relation between auditory filters estimated from psychophysical methods and peripheral tuning was evaluated using a computational auditory-nerve (AN) model that included many of the response properties associated with nonlinear cochlear tuning. The phenomenological AN model included the effects of dynamic level dependent tuning, compression, and suppression on the responses of high-, medium , and low-spontaneous-rate AN fibers. Signal detection theory was used to evaluate psychophysical performance limits imposed by the random nature of AN discharges and by random-noise stimuli. The power-spectrum model of masking was used to estimate psychophysical auditory filters from predicted AN-model detection thresholds for a tone signal in fixed-level notched-noise maskers. Results demonstrate that the role of suppression in broadening peripheral tuning in response to the noise masker has implications for the interpretation of psychophysical auditory-filter estimates. Specifically, the estimated psychophysical auditory-filter equivalent-rectangular bandwidths (ERBs) that were derived from the nonlinear AN model with suppression always overestimated the ERBs of the low-level peripheral model filters. Further, this effect was larger for an 8-kHz signal than for a 2-kHz signal, suggesting a potential characteristic-frequency (CF) dependent bias in psychophysical estimates of auditory filters due to the increase in strength of cochlear nonlinearity with increases in CF. PMID- 11863203 TI - Lymphangioleiomyomatosis: a review of the literature. AB - Lymphangioleiomyomatosis (LAM), characterized by alveolar smooth muscle proliferation and cystic destruction of lung parenchyma, can occur as a rare sporadic disease or as a complication of tuberous sclerosis (TSC). It is a cystic lung disease, usually generalized and progressive, may be extremely difficult to treat and has been considered to have a poor prognosis. It has almost exclusively been reported to present in women of childbearing age, most commonly with dyspnoea and pneumothorax. We reviewed the English literature from 1939 to 1997 for cases of LAM both with and without TSC, in order to document the prevalence, clinical features, investigations, treatment and outcome within and between these two groups. No study has yet determined the prevalence of LAM symptomatically within the general population, but it probably affects 1-3% of the TSC population. Patients with TSC often present with an insidious onset of dyspnoea whilst non-TSC patients present more commonly with acute breathlessness secondary to pneumothorax. Patients with TSC are also less likely to suffer from chylothorax. The age of onset of symptoms and of diagnosis are similar. LAM is rare in children and even less common in males in both groups. The natural course of LAM remains unclear and effect of treatment variable. Although symptomatic LAM is uncommon it causes a significant amount of morbidity and mortality both in the TSC and general population, but asymptomatic LAM is not uncommon in TSC. Further research is required to determine the natural history of this condition and to evaluate current treatment regimes. PMID- 11863204 TI - Determining the optimal number of specimens to obtain with needle biopsy of the pleura. AB - The aim of this study was to define the number of pleural biopsy samples necessary for optimum diagnostic performance and determine to what extent they are complementary. Eighty-four closed pleural biopsies were performed in our department between June 1996 and January 1998 on 55 males and 29 females with an average age of 64.4 +/- 16.7 years. The study of the pleural fluid included: pH, biochemical testing of pleura/serum (proteins, lactate dehydrogenase, glucose, cholesterol, triglycerides, albumin and adenosine deaminase), haemogram, cytology and microbiological testing (Gram-staining, aerobes, anaerobes and mycobacteriae cultures). The biopsies were performed using a Cope needle, with a total of five biopsies for each patient: four for pathological examination (taken numerically in the order in which they were performed: D1, D2, D3 and D4) and one for microbiological testing. In those cases in which the diagnosis was uncertain or effusion persisted, a thoracoscopy or thoracotomy was performed. There were no significant differences in the diagnostic yield of each individual sample (D1, D2, D3 and D4), but there were differences in the sum of the samples, depending on the number of biopsies performed.This was true for total group and the group with carcinomas, but not for the group with tuberculosis. The increase in diagnostic yield with the number of biopsies was more remarkable in the carcinoma cases, where it increased by 35% when four biopsies were performed (54% with one biopsy versus 89% with four biopsies, P < 0.002). In conclusion, the diagnostic yield increased with the number of biopsy samples in the total group and the group with malignancy but not in the group with tuberculous effusions. The best diagnostic performance for malignant pathology was obtained with four samples. In pleural tuberculosis, the diagnostic yield did not increase with more biopsy samples. One high quality sample should be enough to obtain a diagnosis. PMID- 11863205 TI - Prognostic factors of nosocomial pneumonia in general wards: a prospective multivariate analysis in Japan. AB - To determine prognostic factors of nosocomial pneumonia in general wards, we performed prospective clinical study using multivariate statistical analysis. Eighty patients with nosocomial pneumonia in our units were enrolled in the study between December, 1996 and January 1998. Clinical setting and severity of pneumonia were evaluated, and laboratory data were collected at the occurrence of nosocomial pneumonia. Death due to nosocomial pneumonia occurred in 29 of 80 patients (mortality rate 36%). Univariate analysis showed the following factors associated with mortality: the presence of an ultimately or rapidly fatal underlying condition, prior antibiotics use, use of antacids, presence of 'high risk' micro-organisms, sepsis, respiratory failure, multiple organ failure, bilateral chest X-ray infiltrates, a Simplified Acute Physiology Score (SAPS) index > or = 11, albumin < 3.0 g dl(-1), and lactate dehydrogenase (LDH) > or = 796 IUI(-1). Furthermore, multivariate analysis identified three factors significantly associated with mortality: the presence of an ultimately or rapidly fatal underlying condition [odds ratio (OR)=7.0; 95% confidence interval (CI)=1.2 41.1; P=0.03]; SAPS index > or = 11 (OR=7.6; 95% CI=1.1-51.9, P=0.04); LDH > or = 796 IUI(-1) (OR=28.2; 95% CI=2.0-406, P=0.01). Our study indicates that host factors and disease severity factors are important prognostic factors of nosocomial pneumonia in general wards. PMID- 11863206 TI - Extended NO analysis applied to patients with COPD, allergic asthma and allergic rhinitis. AB - The recommended method to measure exhaled nitric oxide (NO) cannot reveal the source of NO production. We applied a model based on the classical Fick's first law of diffusion to partition NO in the lungs. The aim was to develop a simple and robust solution algorithm with a data quality control feature, and apply it to patients with known alterations in exhaled NO. Subjects with allergic rhinitis, allergic asthma, chronic obstructive pulmonary disease (COPD) smokers and controls were investigated. NO was measured at three expiratory flow rates. An iteration method was developed to partition NO. The airway tissue content of NO was increased in asthma, 144 +/- 80 ppb (P = 0.04) and decreased in smokers, 56 +/- 36 ppb (P = 0.02). There was no difference between subjects with rhinitis, 98 +/- 40 ppb and controls, 98 +/- 44 ppb. The airway transfer rate was increased in allergic asthma and allergic rhinitis, 12 +/- 4 vs. 12 +/- 5 ml sec(-1), compared to controls, 8 +/- 2 ml sec(-1) (P < 0.001). The alveolar levels were no different from controls, 2 +/- 1 ppb. In COPD the alveolar levels were increased, 4 +/- 2 ppb (P < 0.001). Extended NO analysis reveals from where in the respiratory system NO is generated. Hence, this new test can be added to the tools the physician has for the diagnosis and treatment of patients with respiratory disorders. PMID- 11863207 TI - Respiratory rate measurement in adults--how reliable is it? AB - Measurement of respiratory rate (RR) is essential in the evaluation of respiratory disorders. However, the variability in RR measurement in adults has never been adequately assessed. Respiratory rate was measured twice in 245 patients; the two measurements were performed by the same observer in 137 patients, by different observers in 58 patients and simultaneously by different observers in 50 patients. The mean (SD) difference between the first and second measurements was 0.03 (3); 95% limits of agreement-4.86-4.94 breaths min(-1), 5.7-5.7 breaths min(-1), and -4.2 to 4.4 breaths min(-1) for the same observer, different observer and simultaneous observer groups, respectively. The difference in RR measurements did not vary with RR. In conclusions on average, there is very good agreement between observers in RR measurement. Inter-observer variability may account for a difference of up to 6 breaths min(-1). This is relevant when applying clinical prediction rules based on threshold RR values. PMID- 11863208 TI - Prevalence of asthma, allergic rhinitis and eczema among university students in Bangkok. AB - Prevalence of childhood asthma is increasing worldwide including in developing countries such as Thailand. Despite a wide availability of epidemiological data on childhood asthma in Thailand, prevalence of asthma in adults has not been well studied within this community. Objectives of this study were to study prevalence of asthma, allergic rhinitis and eczema in a random group of university students in Bangkok using the standardized written and video questionnaires from the ISAAC phase I protocol. The ISAAC phase I, written and video (International version, AVQ 3.0) questionnaires were administered to 3631 randomly selected university students from six universities in Bangkok. The age range of subjects were between 16-31 years (mean +/- SD= 19.1 +/- 1.7 years). There were 1197 males and 2434 females (ratio = 1:2.03). Data were entered into and analysed by the EpiInfo 4 program. Prevalence of wheeze within the past 12 months and of diagnosed asthma for this group of subjects was 10.1% and 8.8%. Prevalence of allergic rhinitis (nasal with eye symptoms within the past 12 months) was 26.3% and of eczema (rash at typical sites of atopic dermatitis, within the past 12 months) was 9.4%. Presence of rhinoconjunctivitis, gender and age were significant risk factors for asthma with male, younger subjects and those with rhinoconjunctivitis reported more frequent asthma-related symptoms (P < 0.01). As in previous studies which found an increase in asthma among Thai children, this survey indicates that the prevalence of asthma among Thai university students has increased to the same level as in several European countries. A large number of adults residing in Bangkok are suffering from rhinitis-related symptoms, the majority of which probably represents allergic rhinitis. PMID- 11863209 TI - Quality of life in acute exacerbation of chronic bronchitis: results from a German population study. AB - This study reports on data from a study conducted in the Federal Republic of Germany examining the quality of life (QoL) of patients with chronic bronchitis (CB) and its acute exacerbations (AECB). Data from 320 patients were collected at AECB and subsequently during a stable phase (non-AECB) utilizing the St George's Respiratory Questionnaire (SGRQ) and the Nottingham Health Profile (NHP). As expected, the QoL of CB patients was poor, even at non-AECB, with patients reporting lower scores than patients with other chronic conditions. Patients reported significantly poorer QoL at AECB than at non-AECB. After adjusting for the severity of the underlying condition, poorer QoL at AECB was significantly and independently associated with older age, unemployment, increasing BMI, increasing number of prior AECBs, and Anthonisen AECB grade. While younger subjects reported significantly greater deterioration in QoL at AECB, the factors most consistently and independently associated with relative QoL deterioration at AECB were the number of prior AECBs and exposure to air pollution at home. In conclusion, this study highlights the detrimental effect of CB, and in particular AECB, on QoL. The association between QoL and patient reports of previous AECB number and air pollution are consistent with reports from other studies. PMID- 11863210 TI - Eosinophilic airway inflammation as an underlying mechanism of undiagnosed prolonged cough in primary healthcare patients. AB - Prolonged cough is a common problem in patients seen in general practice. Using a simple method of sputum induction and processing of sputum samples, we determined whether eosinophilic airway inflammation could be a cause of undiagnosed prolonged cough. Eighty-two patients who had had cough for more than 1 month were enrolled into the study, in six primary healthcare centres. Patients with known pulmonary disease, including asthma or chronic obstructive pulmonary disease (COPD), or who were known to have another cause of cough, or to have recently suffered from a respiratory infection, were excluded. Fifty-three healthy individuals served as controls. Sputum was induced by inhalation of 3% saline. Inflammatory cells in smears were studied semi-quantitatively. Concentrations of eosinophil cationic protein (ECP), eosinophil peroxidase (EPO), myeloperoxidase (MPO) and human neutrophilic lipocalin (HNL) were determined. Sputum induction proved safe and adequate samples were obtained from 91%. Sputum eosinophilia (eosinophils accounting for more than 5% of all cells in smears) was present in 14 patients with prolonged cough (19%) but in no healthy individual (P=0.001). Five of the 14 individuals (36%) who exhibited sputum eosinophilia appeared to have asthma, while nine of the 14 (64%) did not. Concentrations of ECP and EPO were higher in patients with prolonged cough than in healthy individuals (P=0.02 for ECP; 0.005 for EPO). We conclude that eosinophilic airway inflammation is a fairly common cause of prolonged cough, even in patients not suffering from asthma or COPD, or in whom no other cause of cough is known to be present. Induced sputum samples obtained in health centres can be studied in a central laboratory. Detection of eosinophilic airway inflammation could aid the decision regarding treatment. PMID- 11863211 TI - Effect of oxolamine on cough sensitivity in COPD patients. PMID- 11863212 TI - Lymphangioleiomyomatosis and tuberous sclerosis. AB - Tuberous sclerosis (TSC) is an inherited disorder best known for its association with severe learning difficulties, epilepsy, behavioural problems, skin and renal pathology Lymphangioleiomyomatosis (LAM), characterized by alveolar smooth muscle proliferation and cystic destruction of parenchyma, occurs as an infrequent symptomatic pulmonary complication in TSC and as a very rare sporadic disease in those without signs of TSC. Considered a generalized and progressive cystic lung disease that is difficult to treat with a poor prognosis, it has been reported almost exclusively in women, most commonly presenting with dyspnoea and pneumothorax in those of childbearing age. We investigated the clinical features and prognosis of LAM in patients with TSC including the effects of treatment, stratified by the method of diagnosis of LAM (i.e. histological or radiological). We found histological proof of diagnosis in 10 of 21 patients with TSC and symptomatic lung disease, onset in childhood in four, three males with LAM, individuals with apparently focal disease, great variation in clinical course and no clear treatment benefit. In those with TSC, symptomatic LAM is infrequent but causes a significant morbidity and mortality It was not possible to detect predisposing factors, other than being female. Males with apparent LAM should be rigorously investigated. PMID- 11863213 TI - New challenges in the identification of gastrointestinal stromal tumors and other possible KIT-driven tumors. PMID- 11863214 TI - Tissue protection immunohistochemistry: a useful adjunct in the interpretation of prostate biopsy specimens and other selected cases in which immunostains are needed on minute lesions. AB - Performing immunohistochemical analysis on minute lesions is a challenging task, primarily because they frequently disappear when the paraffin block is recutfor immunostaining purposes. This is a particularly common occurrence with prostate biopsy specimens, in which immunostains for high-molecular-weight cytokeratin commonly are used as an adjunct to H&E examination for aiding in the interpretation of minute "suspicious" lesions. We describe an original method designated tissue protection immunohistochemistry, that allows the performance of high-molecular-weight cytokeratin immunostains (or other immunostains) on previously stained H&E slides. The method described does not require destaining of H&E-stained sections, and it allows the preservation of the H&E stain on adjacent levels that may be present on the same slide. The method described requires that the original H&E-stained sections be placed on adhesive slides, but it has the advantages of eliminating the requirement of a paraffin block for immunostaining and eliminating the need for saving intervening unstained sections for possible immunohistochemical analysis. PMID- 11863215 TI - Spindle cell metaplasia of the thyroid arising in association with papillary carcinoma and follicular adenoma. AB - Spindle cell proliferations of the thyroid have been described in association with reactive processes and aggressive malignant neoplasms. We describe spindle cell proliferations in 10 patients arising in association with papillary carcinoma and follicular adenoma. The spindle proliferations were 0.3 to 3.0 cm in size, constituted from 1% to 95% of the primary neoplasm, and were either admixed with the neoplastic elements or peripherally located within the primary tumor Cytologically, these proliferations showed bland-appearing spindle cells with fine chromatin and subtle nucleoli. Mitoses were rare, and inflammation was minimal. Immunostains showed reactivity with thyroglobulin, indicating their follicular origin. We believe it is important to recognize these metaplastic proliferations and distinguish them from aggressive malignant neoplasms. PMID- 11863216 TI - Immunohistochemical detection of Yersinia pestis in formalin-fixed, paraffin embedded tissue. AB - Yersinia pestis infection usually is limited to lymph nodes (bubo); rarely, if bacteria are aerosolized, pneumonic plague occurs. We developed an immunohistochemical assay using a monoclonal anti-fraction 1 Y pestis antibody for formalin-fixed tissues. We studied 6 cases using this technique. Respiratory symptoms were prominent in 2 cases; histologically, one showed intra-alveolar inflammation, and the other had alveolar hemorrhage and edema. By using the immunohistochemical assay, we found intact Yersinia and granular bacterial antigen staining in alveoli, bronchi, and blood vessels. Of the remaining cases, 2 had septicemia and 2 had a bubo. Pathologic changes included lymphocyte depletion, necrosis, edema, and foamy macrophages in lymph nodes; multiple abscesses in the spleen; fibrin thrombi in glomeruli; and unremarkable lungs. By using the immunohistochemical assay, we identified intact bacteria inside monocytes and granular antigen staining in blood vessels. The immunohistochemical assay provided a fast, nonhazardous method for diagnosing plague. The immunohistochemical assay localizes bacteria, retaining tissue morphologic features, and can help define transmission mechanisms. PMID- 11863217 TI - RANK (receptor activator of nuclear factor kappa B) and RANK ligand are expressed in giant cell tumors of bone. AB - In giant cell tumors of bone (GCTBs), the mesenchymal stromal cells are the neoplastic cells and induce recruitment and formation of osteoclasts (OCs). Studies on recently discovered members of the tumor necrosis factor receptor ligand family have demonstrated a crucial role of RANKL (receptor activator of nuclear factor kappa B [RANK] ligand) expressed by osteoblast/stromal cells and of its receptor RANK expressed by OCs during OC differentiation and activation. OCs typically are present in large numbers in GCTBs, suggesting that these tumors may contain cells expressing factors that stimulate OC precursor recruitment and differentiation. We used immunohistochemical analysis to study RANKL and RANK expression in 5 GCTBs. Multinucleated cells and some mononuclear cells showed strong positive staining with anti-RANK antibodies; RANKL was present in a subset of mononuclear cells that did not express the hematopoietic lineage cell marker CD45, a feature that identified them as mesenchymal tumor cells. Our results suggest that RANKL expression may have a role in the pathogenesis of GCTBs and in the formation of the large OC population present in these tumors. PMID- 11863218 TI - Cytologic features of clear cell sarcoma (malignant melanoma) of soft parts: a study of fine-needle aspirates and exfoliative specimens. AB - We describe the cytologic features of clear cell sarcoma of soft tissue (CCS) in 11 fine-needle aspiration biopsy (FNAB) specimens and 6 exfoliative specimens from 11 patients. In 3 patients, FNAB was the initial method of tumor evaluation. In 6 of 11 cases, immunostaining with S-100 or HMB-45 was evaluated. Electron microscopic evaluation was performed in 1 case. Both the FNAB and exfoliative specimens varied in overall cellularity, although reproducible cytologic features were identified. A significant diagnostic pitfall, namely the potential of CCS to form microacinar structures mimicking adenocarcinoma, is described with particular reference to CCS metastatic to regional lymph nodes. A rare case of the granular cell variant of CCS is illustrated as well. Owing to the rarity of CCS, the diagnosis on cytologic smears is extremely difficult and is aided substantially by pertinent clinical data. The diagnosis can be made conclusively by PMID- 11863219 TI - Caveolin-1 expression in ovarian carcinoma is MDR1 independent. AB - We studied the role of caveolin-1 in tumor progression and prognosis in serous ovarian carcinoma and the association between caveolin-1 and MDR1 expression. The study involved immunohistochemical analysis for caveolin-1 and P-glycoprotein (P gp) expression in 75 effusions and 90 solid lesions from ovarian and primary peritoneal carcinoma; in situ hybridization for MDR1 messenger RNA (mRNA) expression in 62 effusions and all 90 tumors; and reverse transcription polymerase chain reaction (RT-PCR) for caveolin-1 mRNA expression in 23 effusions. Immunohistochemical analysis localized caveolin-1 to the cell membrane in 43 effusions and 24 tumors. P-gp membrane expression was detected in 14 effusions and 11 tumors; MDR1 mRNA, in 20 effusions and 30 tumors. Caveolin-1 mRNA was expressed in 19 effusions. Caveolin-1 protein expression showed no association with that of P-gp protein or MDR1 mRNA. The expression of all markers was similar in carcinoma cells in pleural and peritoneal effusions. Caveolin-1 is a novel diagnostic marker for effusions; expression is moderately elevated in tumor cells in effusions, possibly owing to altered signal transduction and metabolism in cancer cells at this site. Expression seems MDR1 independent. PMID- 11863220 TI - Analysis of cyclin D1 expression by quantitative real-time reverse transcription polymerase chain reaction in the diagnosis of mantle cell lymphoma. AB - Mantle cell lymphoma (MCL) is characterized by typical morphologic features and the CD5+, CD23-immunophenotype. However, some morphologically typical MCLs are CD23+. A t(11;14)(q13;q32) translocation is frequently found in MCL, leading to overexpression of cyclin D1. We studied the expression of cyclin D1 in 50 small cell non-Hodgkin lymphomas by real-time reverse transcription-polymerase chain reaction. Most cases with typical MCL morphologic features and immunophenotype gave a strong signalfor cyclin D1, whereas most typical chronic lymphocytic leukemias/small lymphocytic lymphomas (CLLs/SLLs) gave weak or no signals. Based on these results, we determined a threshold value for the diagnosis of cyclin D1 overexpressing MCL. Cyclin D1 expression in 17 lymphomas with conflicting data from morphologic examination and immunophenotyping was variable. The concordance of cyclin D1 measurements with morphologic features and immunophenotype in typical cases proves the usefulness of the method. Unexpectedly high values were found in few CLL/SLL cases and in many CD23+ lymphomas with MCL morphologic features. PMID- 11863221 TI - Precursor T-cell acute lymphoblastic leukemia in adults: age-related immunophenotypic, cytogenetic, and molecular subsets. AB - We analyzed the clinicopathologic and molecular findings in 26 adults (age 16-72 years) with T-cell acute lymphoblastic leukemia (T-ALL) and observed features that correlated with age. Patients older than 60 years (n = 5) had a low frequency of hepatosplenomegaly (0 [0%]), anterior mediastinal mass (1 [20%]), and lymphadenopathy (2 [40%]), and completely responded to chemotherapy (4 of 4). The T-ALL in this group commonly expressed myeloid antigens (4 [80%]), had lineage-inappropriate gene rearrangements (2/3 [67%]) and chromosome 2 deletion (3/4 [75%]), and exclusively used the V(III) or V(IV) families of the T-cell receptor (TCR) gamma gene. In comparison, patients 16 to 60 years old (n = 21) more commonly had an anterior mediastinal mass (8 [38%]), hepatosplenomegaly (10 [48%]), and lymphadenopathy (16 [76%]). The tumors in these patients commonly used the TCR gamma gene VI or V(II) families (17/25 total rearrangements [68%]). Myeloid antigen expression (5 [24%]) and lineage inappropriate gene rearrangements (4/15 [27%]) were uncommon. Within this group, CD1a expression correlated with age 28 to 60 years. These results illustrate considerable age related heterogeneity in adult T-ALL, which may reflect differences in tumor cell maturation. PMID- 11863222 TI - Guidelines for interpreting EBER in situ hybridization and LMP1 immunohistochemical tests for detecting Epstein-Barr virus in Hodgkin lymphoma. AB - Histochemical stains demonstrate Epstein-Barr virus (EBV) in approximately 40% of all Hodgkin hymphomas, suggesting a role in tumorigenesis and the potentialfor EBV-targeted therapy. As research progresses, it is important to define criteria for interpreting histochemical stains. Four hematopathologists independently interpreted EBV-encoded RNA (EBER) and latent membrane protein 1 (LMP1) histochemical stains from 40 cases of Hodgkin lymphoma and then reviewed the stains as a group to resolve discrepancies and to develop interpretation guidelines. To call a Hodgkin case EBV-related, the EBER and/or LMP1 signal must be unequivocally present in Reed-Sternberg/Hodgkin (RS/H) cells. The cytologic features and distribution of stained cells should be matched with those on the corresponding H&E-stained slide to help interpret whether the EBER or LMP1 signal is in malignant or reactive cells. The EBER signal is localized to the nucleus, whereas LMP1 is in the cytoplasm and surface membrane. In some cases, only a fraction of RS/H cells express these factors for technical or biologic reasons. Before calling a case EBER-negative, it is essential to show that tumor cell RNA is preserved and available for hybridization. LMP1 staining, although usually strong among all tumor cells in a given case, may alternatively be focal and weak, contributing to false-negative interpretation. EBER and LMP1 assays in combination are more effective than either assay alone for identifying EBV related Hodgkin lymphoma. PMID- 11863223 TI - Analysis of the human herpesvirus 8 (HHV-8) genome and HHV-8 vIL-6 expression in archival cases of castleman disease at low risk for HIV infection. AB - Lymph nodes from 44 patients with Castleman disease (CD) without risk factors for HIV infection were analyzed with polymerase chain reaction (PCR), in situ hybridization (ISH), and immunohistochemical analysis for human herpesvirus 8 (HHV-8) and viral interleukin-6 (vIL-6). PCR detected HHV-8 genome in 2 of 4 cases; ISH detected it in 9 of 16 cases. HHV-8 vIL-6 peptides were detected in 2 of 44 cases. vIL-6- and ISH-positive cells were found in large transformed and small lymphocytes of the follicular mantle, respectively. Of 9 cases of plasma cell (PC) CD that demonstrated HHV-8 genome by PCR or ISH, 1 expressed vIL-6. Clonal populations of PCs in CD by immunohistochemical analysis or immunoelectrophoresis of serum and urine were associated with neuropathy. HHV-8 vIL-6 detection was associated with poor survival and lack of HHV-8 IL-6, with low risk for subsequent lymphoma. Although HHV-8 genome was detected in a considerable number of patients with PC CD, vIL-6 expression was infrequent. Expression of HHV-8 vIL-6 in CD may indicate poor prognosis in patients at risk for lymphoma who may prospectively require more aggressive treatment. The lack of vIL-6 expression in CD with HHV-8 genome suggests that human IL-6 rather than vIL 6 may be the principal pathogenic cytokine. PMID- 11863224 TI - Association of expression of CD44v6 with systemic anaplastic large cell lymphoma: comparison with primary cutaneous anaplastic large cell lymphoma. AB - CD44 is a ubiquitous multifunctional cell surface adhesion molecule family. High expression of the standard form, CD44s (CD44), and its variant form, CD44v6, has been reported to be associated with tumor dissemination in non-Hodgkin lymphoma. To evaluate the potential role of CD44 and/or CD44v6 in different entities of anaplastic large cell lymphoma (ALCL), 30 cases of systemic ALCL (sALCL; 20 cases) and primary cutaneous ALCL (cALCL; 10 cases) were compared for expression of CD44 and CD44v6 by immunohistochemical staining. Expression of CD44v6 also was analyzed with respect to expression of anaplastic lymphoma kinase (ALK) in sALCL. No difference of CD44 expression was noted between sALCL and cALCL In contrast, expression of CD44v6 was found in 18 (90%) of sALCL cases and in 5 (50%) of cALCL cases. There was no correlation between expression of CD44v6 and expression of ALK in sALCL. These results indicate that expression of CD44v6 rather than CD44 correlates with sALCL. Furthermore, these results suggest that CD44v6 and ALK may be independent predictors of risk for the systemic phenotype of ALCL. PMID- 11863225 TI - Fatal cytotoxic T-cell proliferation in chronic active Epstein-Barr virus infection in childhood. AB - Histopathologic features of 5 cases (4 boys and 1 girl; 4-9 years old) with severe chronic active Epstein-Barr virus (EBV) infection are discussed. All patients died within 3 years after disease onset without developing hematolymphoid malignant neoplasms. The pathology specimens (autopsy, 2 cases; multiple organs and tissues obtained by surgery or biopsy, 3 cases) showed polymorphic lymphocytic proliferation in the lymph nodes (4/5) and spleen (3/3), and systemic lymphocytic infiltration of the liver (4/4), lung (2/2), bone marrow (3/4), and kidney (2/2). Skin lesions were noted clinically in 3 of 5 cases. Two cases had coronary artery aneurysm due to lymphocytic vasculitis. The lymphocytes had a characteristic phenotype of cytotoxic T cells expressing CD3, CD8, and cytotoxic molecules, and were negative for CD4. EBV-encoded small nonpolyadenylated RNAs were detected in the nuclei of the lymphocytes, but latent membrane protein 1 and EBNA2 were not seen. In 4 of 4 cases, an oligoclonal growth pattern of EBV was determined after detecting terminal repetitive sequences by Southern blot. In 3 of 3 cases, the lymphocytes did not have T-cell receptor beta or J(H) gene rearrangement. PMID- 11863226 TI - Assessment of CD10 in the diagnosis of small B-cell lymphomas: a multiparameter flow cytometric study. AB - We evaluated the usefulness of multiparameter flow cytometry with cluster analysis in the diagnosis of a series of 100 well-characterized small B-cell lymphomas (SBCLs). The histologic diagnoses in the 100 cases were follicular lymphoma (FL) in 58, marginal zone lymphoma (MZL) in 17, small lymphocytic lymphoma in 15, and mantle cell lymphoma (MCL) in 10. Of the 58 FLs, 57 were CD10 positive (98% sensitivity). The 1 negative case was unusual in that it occurred in the small intestine. However; architectural, cytologic, and immunohistochemicalfeatures were diagnostic of FL. Of 42 other SBCLs, 2 were CD10+ (95% specificity); 1 was a CD5+/cyclin D1 + MCL, and the other was an extranodal MZL. We found that assessment of CD10 expression using multiparameter flow cytometry with cluster analysis is highly sensitive and specific for the diagnosis of FL, validating its usefulness in situations in which adequate tissue is not available for definitive histologic diagnosis. PMID- 11863227 TI - Usefulness of anti-CD117 in the flow cytometric analysis of acute leukemia. AB - We assessed the diagnostic usefulness of adding anti-CD117 to our existing flow cytometric profile in the analysis of 150 consecutive cases of acute leukemia (de novo or relapsed acute myelogenous leukemia [AML], AML arising in myelodysplastic syndrome, blast crisis of chronic myelogenous leukemia [CML], acute lymphoblastic leukemia, acute unclassifiable leukemia, and biphenotypic leukemia). CD117 was expressed on more than 10% of blasts in 64% of de novo AMLs (42/66), 95% of relapsed AMLs (19/20), 75% of AMLs arising from a myelodysplastic syndrome (6/8), and 25% of myeloid blast crisis in CMLs (1/4). CD117 was not expressed in acute lymphoblastic, acute biphenotypic, or unclassified leukemia or lymphoid blast crisis of CML. The specificity, positive predictive value, sensitivity, and negative predictive value of CD117 for AML were 100%, 100%, 69%, and 62%, respectively. CD117 is a specific marker for myeloblastic leukemias. Sensitivity is greatest in French-American-British M2 and relapsed AML. Intensity of CD117 expression is dim. Despite the high specificity and positive predictive value, the addition of anti-CD117 to our panel did not prove essential for the assignment of blast lineage. PMID- 11863228 TI - Therapy-related acute myeloid leukemia/myelodysplasia with balanced 21q22 translocations. AB - The morphologic and immunophenotypic findings of 36 cases of 21q22 acute myeloid leukemia (AML) and myelodysplasia (MDS) were compared, including 14 de novo t(8;21) AMLs, 11 t(8;21) therapy-related AML/MDS cases, and 11 therapy-related AML/MDS cases with other 21q22 balanced translocations [t(n;21)]. Cases were evaluated for the presence of Auer rods, distinct chunky cytoplasmic blast cell granules, promyelocyte increase, cytoplasmic perinuclear clearing (hofs) of blast cells, eosinophil increase, andfeatures of associated trilineage dysplasia. Results of immunophenotyping studies for CD19, CD34, and CD56 expression were compared. Cases of de novo and therapy-related t(8;21) disease shared common morphologic features of chunky cytoplasmic granules, perinuclear hofs, and promyelocyte increases that were not seen consistently in the t(n;21) group of t AML/MDS cases. Immunophenotypic similarities also were observed between the 2 t(8;21) groups. De novo and therapy-related t(8;21) disease, however, differed by the frequent presence of associated dysplasia in both t-AML/MDS groups, which was infrequent in the de novo t(8;21) group. Therapy-related AMI/MDS with t(8;21) shares characteristic morphologic and immunophenotypic features with de novo t(8;21) AML, but frequently also occurs with associated myelodysplastic changes, similar to other therapy-related acute leukemias. PMID- 11863229 TI - Guidelines for the laboratory use of autoantibody tests in the diagnosis and monitoring of autoimmune rheumatic diseases. AB - The Italian Society of Laboratory Medicine Study Group on the Diagnosis of Autoimmune Diseases has generated a series of guidelines for the laboratory diagnosis and monitoring of systemic autoimmune rheumatic diseases intendedfor the use of clinical pathologists and laboratory physicians. These guidelines are based on a systematic review of published works and expert panel discussion and consist of 13 recommendations for antinuclear antibodies, anti-double-stranded native DNA, and antinuclear specific antibodies. To improve analytic performances and help select the most appropriate test for specific autoantibodies, as well as provide education and guidance in the use of these tests, special emphasis is placed on laboratory methods. PMID- 11863230 TI - Rapid spot tests for detecting the presence of adulterants in urine specimens submitted for drug testing. AB - Several adulterants are used to mask tests for abused drugs in urine. Adulterants such as "Klear" and "Whizzies" contain potassium nitrite, and "Urine Luck" contains pyridinium chlorochromate (PCC). The presence of these adulterants cannot be detected by routine specimen integrity checks (pH, specific gravity, and temperature). We developed rapid spot tests for detecting these adulterants in urine. Addition of 3% hydrogen peroxide in urine adulterated with PCC caused rapid formation of a dark brown color. In contrast, unadulterated urine turned colorless when hydrogen peroxide was added. When urine contaminated with nitrite and 2 to 3 drops of 2N hydrochloric acid were added to 2% aqueous potassium permanganate solution, the dark pink permanganate solution turned colorless immediately with effervescence. Urine contaminated with nitrite liberated iodine from potassium iodide solution in the presence of 2N hydrochloric acid. Urine adulterated with PCC also liberated iodine from potassium iodide in acid medium but did not turn potassium permanganate solution colorless. Urine specimens from volunteers and random urine samples that tested negative for drugs did not cause false-positive results. These rapid spot tests are useful for detecting adulterated urine to avoid false-negative drug tests. PMID- 11863231 TI - Cell block cytology of endometrial aspiration. PMID- 11863232 TI - Qualification of ASCUS. PMID- 11863233 TI - Prisoners of hate. AB - The terrorist attacks in New York and Washington, D.C. on September 11, 2001 as well as domestic terrorism in the United States and elsewhere in the world has prompted an analysis of the psychology of the terrorist. The perpetrators' profound sense of being wronged--their values undermined by foreign powers or a corrupt domestic power structure--has cried out for revolution and revenge. The fanatic ideology of the perpetrators has provided the matrix for a progressively more malevolent representation of the oppressors: the Image of the Enemy. Retribution against the Enemy in the form of mass murder of anonymous civilians becomes an imperative. The counterpart of the image of the Enemy is the idealized collective self-image of members of the movement, faction, or cult. The group narcissism of the white supremacists in the United States, the Aum Shinrikyo in Japan, and the Islamic extremists enhance their collective self-image as pure, righteous, and united. While the foot soldiers, as in any war, gain glory through martyrdom, the instigators and leaders have their own personal narcissistic goals (power and prestige) and plan. For the extremist Islamists the ultimate goal has been overthrow of the moderate Islamic governments; for the domestic terrorists, destabilization of the national government and reinstitution of the traditional values. PMID- 11863234 TI - Expectancy-learning and evaluative learning in human classical conditioning: affective priming as an indirect and unobtrusive measure of conditioned stimulus valence. AB - It has been argued that in classical conditioning two processes might be operative. First, one may learn that the conditioned stimulus (CS+) is a valid predictor for the occurrence of the biologically negative or positive event (US; expectancy-learning). Second, one may learn to perceive the conditioned stimulus itself as a negative or positive stimulus, depending on the valence of the event it has been associated with (evaluative learning). Until the present, however, both forms of learning have been investigated using rather different conditioning procedures. Using a differential aversive conditioning preparation with pictures of human faces as CSs and an electrocutaneous stimulus as US, we were able to demonstrate that both forms of learning can co-occur. Moreover, the extent of evaluative learning in this aversive conditioning procedure did not significantly differ from the amount of evaluative learning in an evaluative conditioning procedure with positive and negative adjectives as USs, which was administered to the same participants. In the present study evaluative learning was not only indexed by direct evaluative ratings, but we introduced affective priming as an indirect and unobtrusive, reaction time based measure of stimulus valence. Finally, imagery instructions during acquisition did not facilitate expectancy learning nor evaluative learning. PMID- 11863235 TI - The effects of safety-seeking behavior and guided threat reappraisal on fear reduction during exposure: an experimental investigation. AB - We examined the effects of safety-seeking behavior and guided threat focus and reappraisal on fear reduction during exposure. Participants (N=46) displaying marked claustrophobic fear were randomized to one of three 30-min exposure conditions: (a) guided threat focus and reappraisal; (b) safety-behavior utilization; or (c) exposure only control. Tripartite outcome assessments during a behavioral approach test, along with measures of suffocation and restriction fears were obtained at pre- and post-treatment, and at a 2-week follow-up. Treatment process measures were collected throughout treatment and consisted of indices of fear activation; within and between-trial fear habituation; and suffocation and entrapment expectancies. Measures of safety behavior utilization and attentional focus were also collected to assess the integrity of the experimental manipulations. Consistent with prediction, those encouraged to utilize safety-behaviors during exposure showed significantly more fear at post treatment and follow-up relative to those encouraged to focus and reevaluate their core threat(s) during exposure. Moreover, growth curve analyses of treatment process data analyses revealed that safety-behavior utilization exerted a detrimental effect on between-trial habituation; whereas guided threat reappraisal enhanced between-trial habituation. PMID- 11863236 TI - Expected and reported pain in children undergoing ear piercing: a randomized trial of preparation by parents. AB - This study examined experimentally the effectiveness of preparatory information provided by parents in creating accurate expectations and reducing children's procedural pain. Ear piercing was used as an analogue to minor painful medical procedures. Sixty children, aged 5-12 years, requesting ear piercing and accompanied by their parents, were randomly assigned to a parental information or contact-control condition. Parents in the information group were asked to read their child a description of the procedures and sensations of ear piercing. Parents in the contact-control condition played picture games for the same amount of time. Expected pain was measured on a visual analogue scale (VAS) and the Faces Pain Scale-Revised (FPS-R) before and after the parental information or contact-control procedure. Experienced pain was measured on the same two scales immediately after the ear piercing. Prepared children had more accurate expectations and reported significantly less pain (M=27.3) than non-prepared children (M=49.8). The validity of the measures was supported by strong correlations (r=0.87 to 0.96) between the VAS and FPS-R. The findings suggest that parental provision of preparatory information creates accurate expectations and reduces pain for children. PMID- 11863237 TI - The management of unwanted pre-sleep thoughts in insomnia: distraction with imagery versus general distraction. AB - Insomniacs commonly complain that they are unable to get to sleep at night due to unwanted thoughts, worries and concerns. The present study investigated whether brief training in identifying and elaborating an interesting and engaging imagery task for use during the pre-sleep period can reduce unwanted pre-sleep cognitive activity and sleep onset latency. Forty one people with insomnia were given one of three instructional sets to follow on the experimental night; instructions to distract using imagery, general instructions to distract, or no instructions. Based on previous findings reported by Salkovskis & Campbell (1994) 'Behaviour Research and Therapy 32 (1994) 1' and ironic control theory (Wegner, 1994) 'Psychological Review 101 (1994) 34', it was predicted that (1) "imagery distraction" would be associated with shorter sleep onset latency and less frequent and distressing pre-sleep cognitive activity compared to the "no instruction" group and that (2) "general distraction" would be associated with longer sleep onset latency and more frequent and distressing pre-sleep cognitive activity compared to the "no instruction" group. Support was found for the first but not the second prediction. The success of the "imagery distraction" task is attributed to it occupying sufficient "cognitive space" to keep the individual from re-engaging with thoughts, worries, and concerns during the pre-sleep period. In addition, "imagery distraction" involved a very specific alternative cognitive task hence the operating process was given a feature positive search, conditions where mental control is likely to be achieved. PMID- 11863238 TI - Mother knows best: effects of maternal modelling on the acquisition of fear and avoidance behaviour in toddlers. AB - The present study was conducted to investigate the influence of parental modelling on the acquisition of fear and avoidance towards novel, fear-relevant stimuli in a sample of 30 toddlers. The toddlers were shown a rubber snake and spider, which were alternately paired with either negative or positive facial expressions by their mothers. Both stimuli were presented again after a 1- and a 10-min delay, while mothers maintained a neutral expression. The children showed greater fear expressions and avoidance of the stimuli following negative reactions from their mothers. This was true for both genders although the degree of modelled avoidance was greater in girls than in boys. The strong observational learning results are consistent with views that modelling constitutes a mechanism by which fear may be acquired early in life. PMID- 11863239 TI - Early abuse, psychiatric diagnoses and irritable bowel syndrome. AB - In a population of 71 (57 female, 14 male) IBS patients seeking psychological treatment, we found expected levels of childhood sexual and physical abuse (57.7%) and expected levels of current Axis I psychiatric disorders (54.9%). Moreover, we found those who had been victims of early abuse had higher current Beck Depression Inventory scores. However, contrary to expectations, there were no significant associations between early abuse and current psychiatric disorder in this population, suggesting that those individuals with psychological distress are not exactly the same group with a history of abuse. PMID- 11863240 TI - An expanded childhood anxiety sensitivity index: its factor structure, reliability, and validity in a non-clinical adolescent sample. AB - Anxiety sensitivity refers to the fear of anxiety-related bodily sensations that are interpreted as having potentially harmful somatic, psychological, or social consequences. The current study examined the factor analytic structure of anxiety sensitivity in a large sample of normal adolescents (N=518) using the revised childhood anxiety sensitivity index (CASI-R). Confirmatory factor analysis indicated that anxiety sensitivity as measured by the CASI-R can best be conceptualised as a hierarchical construct with four lower-order factors loading on a single higher-order factor. The lower-order factors were 'fear of cardiovascular symptoms', 'fear of publicly observable anxiety reactions', 'fear of cognitive dyscontrol', and 'fear of respiratory symptoms'. An additional aim of the present study was to investigate the psychometric properties of the CASI R. Results showed the CASI-R to be a reliable scale in terms of internal consistency. Furthermore, CASI-R scores were substantially related to levels of anxiety sensitivity as measured by the original index, trait anxiety, symptoms of anxiety disorders, in particular 'panic disorder and agoraphobia', and depression. Finally, some evidence was found for the validity of the CASI-R factor scores. That is, all factors convincingly loaded on symptoms of 'panic disorder and agoraphobia', whereas the factor 'fear of publicly observable anxiety reactions' was also strongly associated with symptoms of 'social phobia'. PMID- 11863241 TI - Confirmatory factor analysis of the Penn State Worry Questionnaire. AB - Worry is often assessed with the 16-item Penn State Worry Questionnaire (PSWQ; Meyer, Miller, Metzger, & Borkovec, 1990. Behaviour Research and Therapy, 28, 487 495), but the factor structure of the PSWQ is not well articulated. Three factor analyses of the PSWQ have been conducted, with two (Brown, Antony, & Barlow, 1992. Behaviour Research and Therapy, 30, 33-37; van Rijsoort, Emmelkamp, & Vervaeke, 1999) retaining a unifactorial solution and one (Stober, 1995) retaining a two-factor solution. We sought to determine the relative strengths of these solutions. Seven hundred eighty-eight undergraduates completed the PSWQ and measures of depression and anxiety. Confirmatory factor analyses indicated that the two-factor solution provided a better fit to the data. There was also evidence for a higher order worry factor. Multiple regression analyses revealed that factor 1 (Worry Engagement) explained the majority of the variance in the symptom measures. The higher order worry factor also accounted for variance in some measures. Compared to the PSWQ total score, Worry Engagement demonstrated slightly higher internal consistency and significantly stronger correlations with most measures of depression and anxiety. Factor 2 (Absence of Worry) demonstrated moderate negative correlations with Worry Engagement and the PSWQ Total Score but only modest correlations with depression and anxiety. PMID- 11863242 TI - Cognitive therapy versus interoceptive exposure as treatment of panic disorder without agoraphobia. AB - Cognitive therapy (CT) and interoceptive exposure (IE) as treatments of panic disorder without agoraphobia were compared in a sample of 69 patients, randomly allocated to condition. There were no significant differences between treatments as to reductions in panic frequency, daily anxiety levels and a composite questionnaire score, at posttest after the 12-session treatment, and at both follow-ups (4 weeks, 6 months). In both conditions, high percentages of patients were panic free at post and follow-up tests (range 75-92%). Although the reduction in idiosyncratic beliefs about the catastrophic nature of bodily sensations was equally strong in both conditions, post-treatment beliefs correlated strongly with symptoms at post and follow-up tests in the CT condition, but not in the IE condition. Reduction of beliefs may be essential in CT, but not in IE. This suggests that the two treatments utilize different change mechanisms. PMID- 11863243 TI - Reproducibility of home, ambulatory, and clinic blood pressure: implications for the design of trials for the assessment of antihypertensive drug efficacy. AB - BACKGROUND: The aims of this study were to compare the reproducibility of blood pressure (BP) measured in the clinic (CBP), at home (HBP), and by ambulatory monitoring (ABP), and to assess its implications on the accuracy of antihypertensive drug trials. METHODS: A total of 133 untreated subjects with elevated CBP were assessed with repeated measurements of CBP (five visits within 3 months), HBP (6 workdays within 2 weeks), and ABP (twice, 2 weeks apart). The reproducibility of CBP (one visit), HBP (2 days), and ABP (24 h) was quantified using the SD of differences (SDD) between repeated measurements. The number of subjects required in a comparative trial of two drugs was calculated for each measurement method. RESULTS: We found that HBP provided the lowest SDD values (6.9/4.7 mm Hg, systolic/diastolic, compared with 8.3/5.6 for ABP and 11.0/6.6 for CBP). For a parallel trial aiming to detect a difference in the effect of two drugs of 10 mm Hg systolic BP, 51 subjects would be required when using CBP compared with 29 using ABP and 20 using HBP (73, 53 and 37 subjects, respectively, for the detection of a 5 mm Hg difference in diastolic BP). CONCLUSIONS: The study shows that HBP seems to have superior reproducibility compared with both CBP and ABP. In addition, HBP can improve the accuracy of antihypertensive drug trials, thereby reducing the sample size required. PMID- 11863245 TI - Nighttime blood pressure dipping: the role of the sympathetic nervous system. AB - There is a marked diurnal variation in blood pressure (BP), with BP dipping to its lowest levels during nighttime sleep. A day-night dip in systolic BP (SBP) of <10% has been used to characterize individuals as nondippers, and is associated with an increased risk for cardiovascular disease. The present study examined the contribution of the sympathetic nervous system (SNS) to BP dipping in a biracial sample of 172 men and women aged 25 to 45 years. Assessments included 24-h ambulatory BP monitoring and both waking and sleeping urinary catecholamines. In addition, cardiovascular alpha- and beta-adrenergic receptor (AR) responsiveness was determined by the doses of isoproterenol and phenylephrine required to attain an increase in heart rate of 25 points (CD25) and BP (PD25), respectively. Compared with dippers (n = 116), nondippers (n = 56) were more likely to be African American and to have a family history of hypertension as well as a higher body mass index (BMI). The nighttime fall in both norepinephrine (NE) and epinephrine (EPI) excretion rates was reduced in nondippers compared with dippers (NE dip 9.3 v 13.1 microg/mg; EPI dip 2.7 v 4.0 microg/mg; both P < .05). Nondippers also were characterized by heightened alpha1-AR responsiveness compared with dippers (PD25 = 252 v 321 microg, P < .05). These data suggest that the SNS may contribute to individual differences in nighttime BP dipping, and appears to account in part for blunted BP dipping in African Americans. PMID- 11863244 TI - Effect of amlodipine compared to atenolol on small arteries of previously untreated essential hypertensive patients. AB - In a previous retrospective study, long-term treatment of essential hypertensive patients with a slow-release calcium channel blocker resulted in normal resistance artery structure and endothelial function, which did not occur with a beta-blocker. In the present prospective study, 19 previously untreated essential hypertensive patients (aged 47 +/- 2 years, 75% male) were treated for 1 year in a double-blind randomized study with the long-acting calcium channel blocker amlodipine or the beta-blocker atenolol. Resistance arteries (lumen diameter, 150 to 350 microm) dissected from gluteal subcutaneous biopsies were studied on a pressurized myograph. Blood pressure (BP) control (129 +/- 2/85 +/- 2 mm Hg) was identical in both groups for the last 6 months of the study. After 1 year of treatment with amlodipine, the media-to-lumen ratio (M/L) of resistance arteries decreased from 7.89% +/- 0.40% to 6.81% +/- 0.41% (P < .05). Acetylcholine induced endothelium-dependent relaxation tended to improve from 84.3% +/- 5.5% to 90.5% +/- 4.8% (P = .06), whereas sodium nitroprusside-induced relaxation was unchanged in the patients treated with amlodipine. In the beta-blocker-treated group there was no significant change in M/L or acetylcholine-induced relaxation. In conclusion, treatment with the calcium channel blocker amlodipine corrected altered resistance artery structure and tended to improve endothelial function in essential hypertensive patients, whereas similar good control of BP with the beta blocker atenolol did not. Whether the vascular protective effect of amlodipine will result in improved outcomes in hypertension remains to be demonstrated. PMID- 11863246 TI - Neurovascular contact and blood pressure response in young, healthy, normotensive men. AB - BACKGROUND: Neurovascular contact (NVC) of the left rostral ventrolateral medulla has been implicated in the pathogenesis of "essential" hypertension, and recent studies suggest that this anomaly may be genetically determined. We therefore assessed the prevalence of this vascular anomaly in young normotensive volunteers. We also studied blood pressure, heart rate reactivity, and changes in baroreflex sensitivity in response to mental and physical stress in a subset of subjects with positive and negative brainstem findings. METHODS: Magnetic resonance imaging of the brainstem was performed in 113 young (aged 25 +/- 8 years), normotensive male volunteers. Baroreflex sensitivity was then assessed in 13 subjects with positive brainstem findings and 20 subjects with negative findings. RESULTS: Left-sided NVC was found in 19 subjects. Blood pressure levels, heart rate, and baroreflex sensitivity were similar in all groups. However, modulation of baroreflex sensitivity was reduced under mental and physical stress in subjects with a positive finding for NVC. Subjects with a positive finding also had a significantly lower body mass index than those with a negative finding. CONCLUSIONS: Left-sided NVC is present in approximately one fifth of young normotensive men, and may modulate the baroreceptor reflex under stress in these individuals. PMID- 11863247 TI - Effects of alcohol restriction on ambulatory blood pressure, heart rate, and heart rate variability in Japanese men. AB - We investigated the effects of alcohol restriction on ambulatory blood pressure (BP), heart rate, and heart rate variability in 33 Japanese male volunteers (37 +/- 1 years, mean +/- SE), who were all habitual drinkers. Subjects were told either to keep their usual drinking habits for 3 weeks (usual alcohol period), or to reduce alcohol intake by at least half of their usual drinking amount (reduced alcohol period). The ambulatory BP, heart rate, and electrocardiographic R-R intervals were measured during a 24-h period with a portable recorder on the last day of each period. A power spectral analysis of R-R intervals was performed to obtain the low-frequency (LF) and high-frequency (HF) components. The percentage of differences between adjacent normal R-R intervals >50 msec (pNN50) was also calculated. The amount of ethanol intake was significantly reduced from 70 +/- 5 mL/day in the usual alcohol period to 19 +/- 3 mL/day in the reduced alcohol period (P < .0001). The daytime systolic BP was significantly lower in the reduced alcohol period than in the usual alcohol period by 4 +/- 1 mm Hg (P < .05). The daytime and nighttime heart rate was significantly lower in the reduced alcohol period than in the usual alcohol (P < .001 for each). The pNN50 and the HF component were significantly higher in the reduced alcohol period than in the usual alcohol period (P < .0001 for each). The LF/HF ratio was significantly lower in the reduced period than in the usual period (P < .01). These results demonstrate that 3-week alcohol restriction produced reductions in ambulatory systolic BP, heart rate, and the index of sympathovagal balance, and augmentations of parasympathetic indices of heart rate variability in Japanese male drinkers. PMID- 11863248 TI - Serum potassium is not associated with blood pressure tracking in the Framingham Heart Study. AB - BACKGROUND: Abnormal potassium homeostasis accompanies many secondary forms of hypertension as well as uncommon inherited, monogenic forms of hypertension. We hypothesized that serum potassium may be associated with longitudinal tracking of blood pressure (BP) and development of hypertension. METHODS: A total of 2358 participants (1292 women, 1066 men) in the Framingham Heart Study who were free of hypertension, were not taking drugs affecting potassium homeostasis, and had serum potassium measured in 1979 to 1983 were followed for longitudinal tracking of BP and development of hypertension at examination 4 years later. Progression of BP stage was defined as an increment of one or more BP category, as defined by the sixth report of the Joint National Committee on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure (JNC-VI), between baseline and follow-up examinations. RESULTS: At baseline, there were no differences in systolic or diastolic BP across serum potassium quartiles. Over 4 years of follow up, 37% (457 women, 412 men) of subjects progressed by one or more JNC-VI BP category. In a logistic regression model adjusting for multiple confounders, serum potassium quartile was not associated with risk of BP progression. During follow-up, 14% (162 women, 175 men) of subjects progressed to hypertension. After adjustment for multiple confounders, there was no significant association between serum potassium quartile and risk for progression to hypertension. CONCLUSIONS: In our community-based study sample, serum potassium was not associated with current BP, longitudinal BP tracking, or progression to hypertension. PMID- 11863249 TI - Gender difference in diet-induced obesity hypertension: implication of renal alpha2-adrenergic receptors. AB - Although the pathogenesis of the obesity-related hypertension is not fully understood, prevalence of the cardiovascular complications is much higher in obese men than obese women. In a recent study, we reported that male rats fed a cafeteria diet, while becoming obese, developed hypertension and important changes in their renal alpha2-adrenergic receptor subtypes distributions. The aim of the present study was to investigate whether these alterations are sex dependent. After 10 weeks of the cafeteria diet, male and female rats had the same increase in fat pad weight and in plasma leptin levels. However, in contrast to males, females had normal blood pressure (BP). On the basis of radioligand binding studies using [3H]-RX821002 and confirming our recent observation, an increase in alpha2-adrenergic receptor densities occurred in kidneys of cafeteria fed male but not female rats. Moreover, in contrast with the situation observed in males, ligand competition studies failed to reveal any change in the renal alpha2A- and alpha2B-adrenergic receptors subtypes distribution in females. Finally, in the cafeteria-fed females reverse transcription-polymerase chain reaction experiments showed unaltered expression of these two alpha2-adrenergic receptor subtypes. These data thus suggest a strong relationship between the sexual dimorphism in the cafeteria diet-induced hypertension and altered expression of the alpha2-adrenergic receptor subtypes in the kidney. PMID- 11863250 TI - Salt-sensitive hypertension in conscious rats induced by chronic nitric oxide blockade. AB - This study examined the effects of alterations in salt-intake on blood pressure (BP) in rats chronically treated intravenously with or without the nitric oxide synthase (NOS) inhibitor N(G)-nitro-L-arginine methyl ester (L-NAME) (8.6 mg/kg/day). The changes in mean arterial pressure (MAP), the renal cortical and medullary blood flow (CBF and MBF), and the sodium balance were determined by implanted optical fibers and laser-Doppler flow measurement techniques in the conscious rats. The results showed that high salt intake (7.4 mEq/day) elevates CBF (139% +/- 15%), but has no significant effect on MAP or MBF in control rats; in L-NAME-treated rats, high salt intake elevates MAP, produces no change in CBF, and decreases MBF (51% +/- 14%), as well as increasing the sodium balance (0.26 +/- 0.23 mEq/day to 1.29 +/- 0.47 mEq/day). The present experiments indicated that NO appears to maintain the MBF during high salt intake and to prevent the changes in MAP, and, in the absence of NO, salt-sensitive hypertension develops. Nitric oxide plays an important role in the development of salt-sensitive hypertension with the change of MBF. PMID- 11863251 TI - Reversal of endocardial endothelial dysfunction by folic acid in homocysteinemic hypertensive rats. AB - The role of L- and D-isomers of homocysteine (Hcy) in vascular versus endocardial endothelial (EE) remodeling and function is not well understood. The hypothesis is that Hcy decreases EE cell density by activating matrix metalloproteinase (MMP) and by inducing left ventricular hypertrophy (LVH) in homocysteinemic hypertensive rats (HHR). And L- and D-isomers of Hcy have differential effects in vessel and myocardium. We used: 1) spontaneously hypertensive rats (SHR) in which endogenous total homocyst(e)ine (tHcy) levels are moderately high (18 micromol/L); 2) control age- and sex-matched normotensive Wistar rats (NWR) in which tHcy levels are normal (4 micromol/L); to create hyperhomocyst(e)inemia, 32 mg/day Hcy was administered for 12 weeks in 3) SHR (SHR-H), and in 4) NWR (NWR-H) rats; 5) endogenous tHcy levels were reduced (from 18 to 12 micromol/L) in SHR by folic acid administration (SHR-F). Plasma tHcy levels were measured by HPLC and spectrophometric methods. The MMP activity, measured by zymography, is increased by chronic Hcy administration, and folic acid treatment decreases MMP activity. The collagen and transforming growth factor-beta1 (TGF-beta1), measured by reverse transcriptase-polymerase chain reaction, are increased by Hcy. Folic acid treatment decreases collagen expression and increases TGF-beta1. In vivo LV function was measured in anesthetized rats by a catheter in the left ventricle. The partial decrease in tHcy levels and no change in arterial pressure in SHR after folic acid administration, suggested that folic acid decreases one of the L or D-isomer of Hcy, which is not responsible for an increase in arterial pressure, but may be responsible for myocardial dysfunction. The chronic Hcy administration decreases EE function in NWR and SHR. The treatment of folic acid in SHR improves LVH and EE function. Folic acid improves cardiac remodeling and EE function by decreasing one of the D- or L-isomer of Hcy and by decreasing MMP activity in HHR. These results may suggest a differential role of L- and D isomers in vascular versus cardiac remodeling. PMID- 11863252 TI - Cardiac and vascular fibrosis and hypertrophy in aldosterone-infused rats: role of endothelin-1. AB - Increased endothelin-1 (ET-1) or aldosterone may be associated with promotion of cardiovascular hypertrophy and fibrosis. We evaluated whether the selective ETA receptor-antagonist BMS 182874 (BMS) prevents cardiac and vascular collagen deposition and hypertrophy in aldosterone-infused rats. Rats received subcutaneous aldosterone (0.75 microg/h) and 1% sodium chloride in drinking water +/- BMS (40 mg/kg per day in food) for 6 weeks. Heart and aorta were cross sectioned and stained with Sirius red. Heart weight did not change with either aldosterone infusion or BMS treatment. Cardiac and aortic interstitial and perivascular collagen were quantified with videomorphometry. Aortic collagen and media cross-sectional area were significantly increased 3.5-fold (P < .01) and 1.13-fold (P < .05), respectively, with aldosterone infusion and decreased in BMS treated rats (P < .05, P < .001, respectively). Aldosterone infusion increased interstitial and perivascular collagen in the left (1.6- and 2.7-fold, P < .05 and P < .01, respectively) and right ventricle (1.5- and 1.7-fold, P > .05 and P < .05, respectively). BMS prevented collagen deposition except for interstitial collagen in the right ventricle. Cardiac and aortic fibrosis were significantly increased in aldosterone-infused hypertensive rats. The ETA receptor antagonist prevented cardiac and aortic collagen deposition and aortic hypertrophy. This suggests a role for ET-1 in fibrosis of heart and large vessels in conditions associated with mineralocorticoid excess. PMID- 11863253 TI - Tumor necrosis factor-alpha-induced hypertension in pregnant rats results in decreased renal neuronal nitric oxide synthase expression. AB - BACKGROUND: Preeclampsia is associated with increases in plasma levels of tumor necrosis factor-alpha (TNF-alpha), a cytokine known to contribute to endothelial dysfunction. We recently reported that a twofold elevation in plasma TNF-alpha produces significant reductions in renal function and hypertension in pregnant rats. The purpose of this study was to determine the role of the nitric oxide (NO) system in TNF-alpha-induced hypertension in pregnant rats. METHODS: Tumor necrosis factor-alpha (50 ng/day) was chronically infused starting at day 14 of gestation. Mean arterial pressure, 24-h urinary nitrite/nitrate excretion, and renal nitric oxide synthase (NOS) protein expression by Western blot analysis was measured at day 19 of gestation. RESULTS: A twofold increase in plasma TNF-alpha levels in pregnant rats resulted in a significant increase in arterial pressure (97 +/- 3.6 v 116 +/- 2.1 mm Hg, pregnant versus TNF-alpha pregnant, respectively, P < .05), but no significant change in urinary nitrite/nitrate excretion (22.0 +/- 1.9 v 20.8 +/- 2.5 micromol/24 h, pregnant versus TNF-alpha pregnant, respectively), a measure of whole body NO production. As abnormalities in renal production of NO would not be reflected in the measure of whole body NO production, changes in renal NOS protein levels were determined. The protein expression of both neuronal (nNOS) and inducible (iNOS) nitric oxide synthase were significantly decreased in the medulla of TNF-alpha pregnant rats (nNOS: 10.6 +/- 0.7 v 8.2 +/- 0.8 densitometric units, P < .05; and iNOS: 19.2 +/- 0.9 v 15.4 +/- 0.8 densitometric units, P < .05, pregnant versus TNF-alpha pregnant, respectively). CONCLUSION: The hypertension associated with a chronic twofold increase in TNF-alpha in pregnant rats is associated with significant decreases in renal nNOS and iNOS protein production. PMID- 11863254 TI - Endothelial dysfunction in cold-induced hypertensive rats. AB - Endothelial dysfunction can be observed in preatherosclerotic conditions. However, its pathogenetic role in hypertension is still controversial. Endothelial-dependent changes of blood pressure (BP) and expression of endothelial nitric oxide synthase (eNOS) were evaluated in cold-induced hypertensive rats. Wistar rats were exposed to cold stress for 8 weeks. Exposure to cold stress significantly increased the systolic BP in rats. The infusion of acetylcholine significantly lowered mean arterial BP in control rats by 48 +/- 2% and by 32 +/- 1% in cold-induced hypertensive rats. The acetylcholine-induced reduction of mean arterial BP was significantly attenuated in cold-induced hypertensive rats (control rats, 45 +/- 2 mm Hg; cold-induced hypertensive rats, 34 +/- 3 mm Hg; P < .05). Administration of N(G)-nitro-L-arginine-methyl ester for 1 week significantly increased BP in control rats, whereas no effect could be observed in cold-induced hypertensive rats. In cold-induced hypertensive rats eNOS in aortic vessels was significantly reduced compared to control rats. In this nongenetic, nonsurgical animal model of cold-induced hypertensive rats an endothelial dysfunction can be observed due to reduced eNOS. PMID- 11863255 TI - Comparative effects of the dual ACE-NEP inhibitor MDL-100,240 and ramipril on hypertension and cardiovascular disease in endogenous angiotensin II-dependent hypertension. AB - We investigated the effects of MDL-100,240 in a transgenic rat model (TGRen2) of hypertension with severe cardiovascular damage (CVD) due to enhanced tissue synthesis of angiotensin II (Ang II). Male heterozygous TGRen2 rats (5 weeks old) were allocated to receive MDL-100,240, ramipril (RAM) or placebo (PLAC) for 4 weeks, during which blood pressure (BP) was measured. We then evaluated: 1) left ventricle (LV) and right ventricle (RV), brain, kidney and adrenals weight; 2) structural changes in the aorta and the mesenteric arterioles wall; 3) tension responses of segments of the aorta to phenylephrine, KCl, and endothelin-1; and 4) creatinine, aldosterone, atrial natriuretic peptide (ANP), and cyclic GMP (cGMP) plasma levels. Compared to PLAC, both MDL-100,240 and RAM significantly (P < .001) lowered BP (after 4 weeks: 255 +/- 15 mm Hg PLAC, v 174 +/- 6 MDL 100,240, v 166 +/- 5 RAM). They hindered LV hypertrophy (3.73 +/- 0.25 mg/g body weight (PLAC) v 2.71 +/- 0.22 (MDL-100,240) P < .001; v 2.36 +/- 0.2 (RAM), P < .001). MDL-100,240 also prevented aortic dilatation and hypertrophy of the mesenteric arterioles (media thickness, 25.3 +/- 0.5 microm PLAC, v 21.1 +/- 0.9 MDL-100,240, P < .007; v 20.2 +/- 1.5 RAM, P = .033) and lowered the tension responses to phenylephrine (P < .01), KCl (P < .01), and endothelin-1 (P < .001). Plasma aldosterone (710 +/- 153 pmol/L PLAC, v 237 +/- 61 MDL-100,240, v 180 +/- 22 RAM) and creatinine levels (0.69 +/- 0.33 mg/dL PLAC, v 0.41 +/- 0.02 MDL 100,240, v 0.41 +/- 0.04 RAM) were also decreased (P < or = .001). Compared to PLAC, plasma ANP levels were 11% and 2.4% higher in MDL-100,240 and RAM, respectively (both P = not significant); cGMP levels were unaffected. Thus, severe hypertension and related CVD were regressed by MDL-100,240, which resulted to be as effective as a full dosage of ramipril in TGRen2. PMID- 11863256 TI - Genotypes of the betaENaC gene have little influence on blood pressure level in the Japanese population. AB - The gene for the beta-subunit of the epithelial sodium channel (betaENaC) is one of the most prominent candidate genes being analyzed for an association with human essential hypertension. It is known that a deletion or alteration of PY motif in exon 12 of betaENaC is responsible for Liddle's syndrome. Although the localization of genetic polymorphisms of betaENaC is unique to each population, intensive analysis of individuals of white and African ancestry has demonstrated that genetic variants are localized in exons 8 and 12, with two frequent polymorphisms, G442V in exon 8 and T594M in exon 12. These two mutations are both found in individuals of African ancestry, and might be associated with elevated blood pressure (BP). Previously, we have screened the last two-thirds of exon 12 in the Japanese population, and demonstrated the absence of the T594M mutation and the presence of a novel P592S mutation. In the present study, we further examined the rest of exon 12 and exon 8 in a general population from Ohasama, Japan (the Ohasama Study), using single-strand conformational polymorphism (SSCP) analysis. We screened 803 subjects randomly selected from the representative participants, who measured their home and casual BP. The PCR products presenting a shift in SSCP gels, as well as controls, were directly sequenced by autoanalyzer to identify the mutation. A novel gel shift was noted in exon 12 (n = 8) and sequencing identified a polymorphism at codon Ser 520, leading to no change in amino acid sequence (G77576C TCG-->TCC). In exon 8, all three SSCP variants were heterogynous for V434M (GTG-->ATG), which is coincident with a rare polymorphism in whites. The G442V mutation, however, was absent from the Japanese population. A novel mutation of exon 12 was not associated with a significant difference in clinical features. These results indicate that Japanese people possess three polymorphisms in exon 12, all of which are unique, and one in exon 8. These genetic variants of betaENaC may not influence the BP level of Japanese people. PMID- 11863257 TI - Design of the Omapatrilat in Persons with Enhanced Risk of Atherosclerotic events (OPERA) trial. AB - The Omapatrilat in Persons with Enhanced Risk of Atherosclerotic events (OPERA) trial is a large clinical trial of omapatrilat, a vasopeptidase inhibitor, in patients with stage 1 isolated systolic hypertension (ISH). OPERA is the first study to examine whether effective antihypertensive treatment can provide survival and clinical end point benefits in older persons with this common condition. This 5-year multinational, randomized, double-blind, parallel-group, placebo-controlled, forced-titration study will be conducted in approximately 12,600 subjects randomized by approximately 1100 study centers worldwide over a recruitment period of approximately 2 years. The primary objective of OPERA is to determine whether treatment with once-daily omapatrilat (target dose 40 mg) will reduce cardiovascular (CV) morbidity and mortality in older (> or = 65 years) men and women with enhanced risk for atherosclerotic events due to stage 1 ISH plus other risk factors for which currently there is no evidence-based requirement for treatment. Blood pressure inclusion criteria are systolic blood pressure (SBP) 140 to 159 mm Hg (SBP 125 to 139 mm Hg in diabetic individuals) and diastolic blood pressure (DBP) <90 mm Hg. The primary end point is defined as the composite of fatal/nonfatal stroke, fatal/nonfatal myocardial infarction, fatal/nonfatal heart failure, and other CV mortality. Secondary end points include the individual components of the primary end point, CV mortality, and major cardiovascular end points, as well as effects on cognitive function and initiation of treatment for diabetes. Additional analyses will be conducted in men and women, in diabetic patients, in different risk classes and according to prior evidence of vascular disease. PMID- 11863258 TI - Resistant hypertension in a young man with asthma. PMID- 11863259 TI - Power and replication in case-control studies. AB - The case-control study design, a common staple of epidemiology, is increasingly used to test for genetic association. The simplicity of the design accounts for both its appeal and its limitations. Too often, however, apparent controversy arises for lack of appreciation of basic tenets underlying statistical testing. Power and replication are two concepts most commonly ignored in evaluating such studies. We review the basic principles of statistical testing, recall simple means to calculate power, and provide numerical examples pertaining to the association between angiotensinogen and essential hypertension. PMID- 11863260 TI - Effect of fluvastatin and pravastatin, HMG-CoA reductase inhibitors, on fluconazole activity against Candida albicans. AB - Synergy between fluvastatin, at clinically unachievable concentrations, and fluconazole against Candida albicans has been reported. The purpose of the present study was to evaluate the in-vitro activity of fluconazole alone and in combination with clinically achievable concentrations of pravastatin and fluvastatin against C. albicans. In-vitro susceptibility and synergy testing were performed against clinical isolates of C. albicans with fluconazole, pravastatin and fluvastatin. Both checkerboard method and time-kill studies were performed. MICs for fluconazole ranged from 0.5 (susceptible) to >256 mg/L (resistant) at 24 h. All isolates had MICs >2 mg/L for both statins. No synergy or antagonism was observed with fluconazole in combination with either agent against any isolate of C. albicans by the checkerboard assay or time-kill studies. Clinically achievable concentrations of pravastatin and fluvastatin did not affect the in-vitro activity of fluconazole against C. albicans. PMID- 11863261 TI - Aetiology, antifungal susceptibility, risk factors and outcome in 201 fungaemic children: data from a 12-year prospective national study from Slovakia. AB - A total of 201 cases of fungaemia in children in a 12-year national survey from seven University Paediatric Clinics in Slovakia in 1990-2001 was assessed to determine risk factors, therapy and outcome, and to compare those cases with fungaemia in 130 adult cancer patients studied in a similar survey. Four univariate analyses were performed to assess differences in aetiology, antifungal susceptibility and outcome between fungaemia in neonates and paediatric intensive care unit (ICU) patients as well as between paediatric and adult cancer patients with fungaemia. There was a significant difference in aetiology and antifungal susceptibility between the subgroups of children with fungaemia: 83.3% of neonates versus 40.2% in children with cancer were due to Candida albicans. None of the non-albicans Candida spp. (NAC) in neonates but 23.5% of NAC isolates from children with cancer were resistant to fluconazole. C. albicans caused 144 (71.1%) episodes and NAC 48 (23.7%) episodes. Trichosporon beigelii, Blastoschizomyces (Trichosporon) capitatus, Rhodotorula rubra and Cryptococcus laurentii were found less frequently in neonates than in children with cancer (18.8%). There were not many differences in risk factors between paediatric fungaemia and adult cancer fungaemia except C. albicans aetiology, corticosteroid use in therapy, breakthrough fungaemia after ketoconazole prophylaxis and meningitis as a complication, which were observed significantly more frequently among children than in adults, both with cancer and fungaemia. Thirty-three of the paediatric fungaemias were breakthrough cases and appeared frequently in children with cancer. Fifty-one (25.1%) children died with fungaemia (attributable mortality) and 25 (12.7%) due to underlying disease with fungaemia; overall mortality was 37.8% and there was no significant difference in death rates between the subgroups of paediatric patients (neonates, children in ICUs and children with cancer). PMID- 11863262 TI - PCR identification of dermatophyte fungi Trichophyton rubrum, T. soudanense and T. gourvilii. AB - Diagnosis of dermatophytosis employing conventional laboratory procedures has been complicated by the slow growth and varied morphological features shown by dermatophytes. After analysis of the nucleotide base sequences of a 1.2-kb fragment amplified from a dermatophyte fungus Trichophyton rubrum by arbitrarily primed PCR with random primer OPD18, a pair of primers (TRIF and TR1R) was designed and evaluated for specific identification of T. rubrum. The sensitivity of the primers TR1F and TR1R was high, as a specific PCR band of c. 600 bp was detected from as little as 7 pg of T. rubrum DNA. By examining 92 dermatophyte strains and clinical isolates, it was found that this pair of primers reacted in PCR with T. rubrum, T. soudanense and T. gourvilii through formation of the specific fragment of 600 bp, but not with any other of the dermatophyte species or varieties, fungi, yeasts or bacteria tested. As T rubrum is one of the most frequently isolated dermatophyte fungi, and T. soudanense and T. gourvilii are relatively uncommon in many parts of the world, these primers can be used for rapid, sensitive and specific identification and differentiation of T. rubrum from other fungi and micro-organisms. PMID- 11863263 TI - RecA and glnA sequences separate the bacteroides fragilis population into two genetic divisions associated with the antibiotic resistance genotypes cepA and cfiA. AB - The sequences of part of the glutamine synthetase-encoding gene (glnA) and of the RecA-encoding gene (recA) were determined and aligned for 45 Bacteroides fragilis isolates from different clinical and geographical origin. The patterns of sequence divergence of glnA and recA were very similar. The sequences of a 303-bp fraction of recA showed 45 nucleotide substitutions, 40 of which allowed the separation of B. fragilis into two major divisions, which were not found when the deduced amino acid sequences were considered. The 687-bp sequences analysed for the glnA gene showed 112 nucleotide substitutions, 96 of which separated the population into the same two divisions as those described for recA. In this case, the deduced amino acid sequences showed this subdivision as well: three of the six observed amino acid substitutions were division-specific. Within the two divisions, both genes presented a high degree of sequence conservation. Each B. fragilis division was associated with the presence of a different antibiotic resistance gene: cepA encoding a serine-beta-lactamase (division I) and cfiA encoding a metallo-beta-lactamase (division II). No particular clusters associated with geographical or clinical origin, or with the production of an enterotoxin were observed. Sequencing of the cfiA gene allowed identification of two different alleles in division II. However, no association of these different cfiA alleles with the expression of imipenem resistance was observed. In conclusion, the phylogenetic patterns observed by sequencing recA and glnA are in agreement with those obtained previously by MLEE (multilocus enzyme electrophoresis). Thus, it appears that the evolution of recA and glnA genes is similar to that of the whole chromosome of B. fragilis. Horizontal gene transfer between divisions I and II seems to be low, at best. However, the results of the present study could not clarify definitively whether divisions I and II should be considered as two different B. fragilis genospecies. PMID- 11863264 TI - Pulmonary granulomas of guinea pigs induced by inhalation exposure of heat treated BCG Pasteur, purified trehalose dimycolate and methyl ketomycolate. AB - This study was designed to determine the identity of granulomatogenic substances in Mycobacterium bovis BCG Pasteur. When heat-treated BCG Pasteur bacilli were introduced into the lungs of guinea-pigs by an inhalation exposure apparatus, pulmonary granulomas without necrosis developed. Furthermore, when four kinds of mycolates derived from M. tuberculosis Aoyama B strain were introduced into the lungs by the same method, only trehalose 6,6'-dimycolate (TDM) and methyl ketomycolate induced pulmonary granulomas without central necrosis. The pulmonary granulomas consisted of epithelioid macrophages and lymphocytes. When a mixture of TDM and anti-TDM antibody was introduced into the lungs, development of granulomatous lesions was reduced. These data indicate that TDM and methyl ketomycolate are potent granulomatogenic reagents. PMID- 11863265 TI - Variations in 10 putative uropathogen virulence genes among urinary, faecal and peri-urethral Escherichia coli. AB - A total of 868 isolates was screened from seven different collections of organisms from previous studies - pyelonephritis in children aged 1-24 months; first, second and recurring urinary tract infection (UTI) in women aged 18-39 years; UTI in women aged 40-65 years and peri-urethral and faecal isolates from women aged 18-39 years - for the presence of 10 potential Escherichia coli UTI virulence genes. Previously reported differences between the frequency of these genes in UTI compared with faecal isolates were confirmed and extended. A single virulence signature (strains containing aer, kpsMT, ompT, fim and papGAD) occurred in 29% of the pyelonephritic isolates, but in no more than 11% of the other collections. Peri-urethral isolates were found to have frequencies of these 10 genes that differed from those found for both UTI and faecal isolates. PMID- 11863266 TI - The interaction of Yersinia pestis with erythrocytes. AB - Human and murine erythrocytes (RBC) were invaded by Yersinia pestis in vivo and in vitro during a short period and were probably used as an essential source of iron and porphyrin for survival, effective gross multiplication and rapid spread of these bacteria in the bloodstream of mammals. Both iron and porphyrin were extracted by Y. pestis from the RBC through oxidase-catalase activity which produced oxidation of the RBC glucose with generation of H2O2 in large concentration leading to oxidative transformation of haemoglobin into haemin. Furthermore, some mainly chromosomally encoded effector proteins were implicated in this process because all were synthesised by Y. pestis grown in media simulating the intracellular conditions of mammalian RBC. Damaged RBC lost the ability to transport O2 in the mammalian organism. As a result, significant oxygen deficiency developed in host tissues providing specific clinical disease features of plague which are similar to characteristic symptoms of poisoning with haemotoxic substances when the transformation of the haemoglobin Fe2+ to Fe3+ occurs. PMID- 11863267 TI - Induction of apoptosis of human macrophages in vitro by Legionella longbeachae through activation of the caspase pathway. AB - The cytotoxicity of the facultative intracellular bacterium, Legionella longbeachae, an important cause of legionellosis, was characterised. Apoptosis was induced in HL-60 cells, a human macrophage-like cell line, during the early stages of infection and induction of apoptosis correlated with cytotoxicity. Apoptosis was confirmed by agarose gel electrophoresis of fragmented DNA, surface exposure of phosphatidylserine and propidium iodide labelling of host cell nuclei. The involvement of macrophage infectivity potentiator (Mip) protein, a known virulence factor of L. longbeachae, was also examined. A mip mutant of L. longbeachae induced apoptosis of HL-60 cells but failed to multiply intracellularly, suggesting that intracellular replication of L. longbeachae is not essential for the induction of apoptosis of HL-60 cells. Furthermore, induction of apoptosis of L. longbeachae-infected macrophages was mediated by activation of the caspase pathway but might be independent of tumour necrosis factor-alpha- and Fas-mediated signal transduction pathways. PMID- 11863268 TI - Detection of enterotoxigenic Clostridium perfringens with a duplex PCR. AB - Two sets of primers designed to detect Clostridium perfringens phospholipase C (plc) and enterotoxin (cpe) genes in a single PCR reaction were applied to a collection of 64 predominantly food poisoning-related C. perfringens isolates. In vitro enterotoxin synthesis was tested serologically after inducing sporulation. Of the 64 isolates, 26 were clearly enterotoxigenic; 16 were classified as potentially enterotoxigenic only as serological testing did not confirm enterotoxin production. Duplex PCR for diagnosis of enterotoxigenic C. perfringens from vegetative cultures can be a useful tool as fresh isolates often sporulate poorly or not all, giving rise to the possibility of false negative results by serological analysis. PMID- 11863269 TI - Rapid identification of Streptococcus intermedius by PCR with the ily gene as a species marker gene. AB - Streptococcus intermedius belongs to the anginosus group of streptococci (AGS) and is associated with endogenous infections leading to abscesses in the oral cavity and at deepseated sites, such as the brain and liver. Two other species, S. anginosus and S. constellatus, and some presently unnamed taxa, are also classified as AGS. Recently, S. constellatus subsp. pharyngis, a new subspecies with biochemical characteristics similar to S. intermedius, was described with the potential for causing confusion when trying to identify isolates of these two species routinely with commercial identification kits, such as Rapid ID32 Strep and Fluo-Card Milleri. To correctly identify S. intermedius, this study attempted to develop an accurate PCR identification system with the ily gene as a species marker. This approach relies on amplification of an 819-bp fragment of the ily gene and its 3'-flanking region and is shown here to be specific for S. intermedius strains among all other streptococcal species. Moreover, this PCR system was applicable in direct rapid PCR with whole bacterial cells and TaKaRa Z Taq (TaKaRa), a highly efficient DNA polymerase, as the template and DNA amplification enzyme, respectively. PMID- 11863270 TI - Meningo-encephalitis in a neonate caused by maternal Mycoplasma hominis treated successfully with chloramphenicol. PMID- 11863271 TI - Amphotericin B and invasive aspergillosis--how do the data guide us? PMID- 11863272 TI - The pneumococcus: carriage, disease and conjugate vaccines. AB - Modern biotechnology has made possible the rapid development and introduction into clinical care of a wide spectrum of potent antimicrobial agents. However, the battle against Streptococcus pneumoniae (pneumococcus) has remained fierce, as acquisition of resistance is even more rapid and these antimicrobial agents are rendered ineffective. Obtaining appropriate antibiotic treatment for severe invasive pneumococcal infections is now a major challenge in many regions of the world. The ground-breaking success of Haemophilus influenzae type b (Hib) conjugate vaccine has brought hope for the conquest of other capsulate bacteria. Recent results of efficacy trials of a heptavalent pneumococcal conjugate vaccine bring hope that protein conjugate vaccines will have a similar impact on pneumococcal disease. These multivalent vaccine formulations include pneumococcal serotypes that most often acquire antibiotic resistance and there is hope that the widespread application of these vaccines will decrease the incidence of multi drug-resistant infections. The potential reduction of pneumococcal disease could even extend to unimmunised younger siblings and the elderly residing with immunised young children, through its herd effect. However, in view of the multiplicity of serotypes and the biology of the pneumococcus, optimism must be tempered by caution. PMID- 11863273 TI - Interest in and obstacles to pursuing work among unemployed dually diagnosed individuals. AB - Employment status is commonly used as a sign of stability in recovery and an outcome variable for substance abuse treatment and research. However, there has been little attention in the literature on the topic of work for the dually diagnosed (i.e., persons diagnosed with both substance use and mental health disorders). Data collected in 1999 are presented on expressed interest in and perceived barriers to pursuing work and on the utilization of vocational rehabilitation (voc-rehab) services among unemployed members of a dual recovery self-help fellowship (N= 130). While members generally expressed high interest in working, they also cited multiple obstacles to attaining and maintaining employment. A path model was specified and tested. Significant contributors to interest in working were substance use status and physical health rating. Consistent with our hypotheses, mental health symptoms and greater perceived obstacles (e.g., stigma, fear of failure, and insufficient skills) were significant contributors to perceived difficulty in pursuing work, whereas substance use, physical health, and recency of employment were not. Finally, those who perceived less difficulty in pursuing work were more likely to utilize voc-rehab services, and men were more likely than women to use these facilities; interest in work was not significantly associated with utilizing voc-rehab services. The roles of mental health disorders and substance use in relation to pursuit of employment are discussed, as well as that of perceived obstacles such as stigma. The paper addresses the setting of realistic vocational goals and possible strategies to mitigate barriers to increased employment of dually diagnosed individuals. PMID- 11863274 TI - Spit tobacco prevention and cessation counseling: statewide survey of health-care professionals and educators. AB - The use of spit tobacco (ST) products is a serious public health problem in the United States. Use of ST is associated with increased risk of oral cancer, gastrointestinal neoplasms, and other deleterious effects. The prevalence of ST use among adolescents is high in many areas, especially in predominantly rural states (e.g., South Dakota, Montana). Community-wide efforts aimed at prevention and cessation of ST use among young people are needed. A total of 4089 clinicians and educators were surveyed in 1998 regarding their personal ST use and several other characteristics associated with ST prevention and cessation counseling. Educators reported significantly higher rates of ST use than did clinicians. The most prevalent barriers to ST counseling among clinicians were perceptions of patient resistance to referral to ST cessation programs and the lack of community services that effectively treat ST use. Lack of training was a major barrier to ST counseling among all educator subgroups. Although knowledge of the health effects of ST was fairly high among all subgroups, more than 10% of dentists and dental hygienists failed to report that ST use causes gum disease. Most clinicians believed that they should demonstrate leadership in efforts aimed at ST control; however, only 64% of dentists believed that repeated counseling attempts were necessary with patients who continued to use ST. Compared with clinicians, educators generally felt less obligated to provide ST counseling. Eighty percent of physicians reported counseling activities, but fewer than half of the dental professionals did. More than 75% of Drug Abuse Resistance Education (DARE) officers reported having delivered ST counseling, whereas only 4% of volunteer leaders did. Fewer than 50% of educators believed that the ST program they taught was effective. Training of various professionals in ST interventions may benefit from emphasizing different issues (reduction of personal ST use, knowledge, commitment, etc.). Our findings have implications for community-based efforts aimed at controlling ST use among young people. PMID- 11863275 TI - Gambling in mild-moderate alcohol-dependent outpatients. AB - There is a growing interest in the comorbidity of "substance use disorder" and "problem gambling." although there has been little study specifically on people with "alcohol dependence" who are being treated in general alcohol- and drug-user outpatient settings. This study aimed to determine the nature and extent of gambling in a sample of 124 mild-moderate alcohol-dependent outpatients. Of these, 79.8% had gambled in the previous 6 months and 29.8% on at least a weekly basis. Although a wide range of gambling modes was used, by far the commonest was Lotto, a national weekly lottery, at 60.5%. Some 19.4% were found to manifest current "problem gambling" [i.e., scored at least 1 on the South Oaks Gambling Screen (SOGS) instrument], and a further 4.0% were found to manifest pathological gambling confirmed by Diagnostic and Statistical Manual of Mental Disorders Version IV (DSMIV) diagnosis. "Problem gamblers" were significantly more likely to be involved in all modes of gambling compared with non problem gamblers. However, the most differentiating modes, in order, were gambling machines, dogs, casino, and horses. Treatment implications of these findings are discussed. A two arm model of intervention for problem gambling within the alcohol- and drug-user treatment setting is proposed. PMID- 11863276 TI - A cross-national study of motives for drinking alcohol. AB - The role of individualism-collectivism in influencing the motives for alcohol use was examined by comparing motives for drinking among 72 U.S. (individualist) and 83 Nigerian (collectivist) respondents in 1998. The study also examined the role of gender. The Drinking Motives Questionnaire (Cooper, M.L.; Russell. M.; Skinner. J.B.; Windle, M. Development and Validation of a Three-Dimensional Measure of Drinking Motives. Psychol. Assess. 1992, 4, 123-132.) that classifies drinking motives into coping. enhancement, and social motives was used. The U.S. respondents were hypothesized to score higher on coping motives and lower on social motives than Nigerian participants. This hypothesis was supported. Male respondents also scored higher than females on all the three motives. These results have implications for intervention approaches regarding alcohol use for people with differing cultural orientations. PMID- 11863277 TI - Factors associated with regular marijuana use among high school students: a long term follow-up study. AB - The present study investigated whether several behavioral and psychosocial factors measured during early adolescence predicted regular marijuana use 6 years later in a sample of high school students. As part of a school-based survey. 7th grade students (N = 1,132) reported levels of alcohol, tobacco, and marijuana use, and were assessed on several domains of psychosocial functioning potentially relevant in the etiology of marijuana use. When students were followed-up in the 12th-grade, 14% smoked marijuana on a regular basis (once or more per month). Findings indicated that early cigarette smoking, alcohol use, and alcohol intoxication predicted later regular marijuana use. For boys, early marijuana use increased the odds for later regular marijuana use. Cigarette smoking by friends and siblings during early adolescence also increased the likelihood of later monthly marijuana use. The findings suggest that early prevention programs for adolescent alcohol, tobacco, and/or other drug use may have important preventive effects in terms of potentially more serious levels of marijuana involvement later in adolescence and early adulthood. PMID- 11863278 TI - AA utilization after introduction in outpatient treatment. AB - Treatment for alcohol dependence is often provided in outpatient settings, and often includes introduction to Alcoholics Anonymous (AA). Relatively little is known about subsequent AA utilization. Analyses of survey data collected from 72 clients of an outpatient treatment center introduced to AA revealed that, 6 months following intake, a large portion of the responding sample of 55 were still attending AA meetings. Principal components analysis of self-reports of the frequencies of 12 AA-related behaviors found three dimensions of AA utilization: fellowship or social involvement, meeting attendance and participation, and involvement in bureaucratic functioning and meeting production. Results suggest it is important to consider these dimensions of utilization for those wishing to understand AA involvement. PMID- 11863279 TI - Feasibility of computerized scheduled gradual reduction for adolescent smoking cessation. AB - The purpose of this project was to modify a smoking cessation program that uses computerized scheduled gradual reduction for use with adolescent smokers and to test the feasibility of this cessation approach in group support and minimal contact modalities. Utilizing a lesson plan approach with high school marketing students in five high schools and student survey feedback, the LifeSign program was modified to be an acceptable smoking cessation program for adolescent smokers. In the first study, 17 adolescent smokers used the modified program with seven associated weekly group support sessions. At the end of treatment, 29% had quit smoking, and over half of those who continued to smoke reduced their smoking rate by 50%. In the second study, the LifeSign for Teens program was evaluated with 18 adolescent smokers in a minimal contact format. At the end of treatment, 17% had quit smoking, and mean smoking rate reductions of 43% were found among those who continued smoking. At 1-year follow-up, all subjects who had quit at posttreatment reported continuous abstinence. The results of these two small trials suggest that a computerized scheduled gradual reduction approach may be an accepted and potentially efficacious approach for smoking cessation among adolescent smokers. PMID- 11863280 TI - Enantioselective analysis of atenolol in biologic fluids: comparison of liquid liquid and solid-phase extraction methods. AB - In this study we evaluated a liquid-liquid extraction procedure and a solid-phase extraction procedure for sample preparation for the enantioselective analysis of atenolol in plasma and urine by high-performance liquid chromatography. A Chiralcel OD-H column was used for the resolution of atenolol enantiomers with hexane-ethanol (85:15, v/v) plus 0.1% diethylamine as the mobile phase. In the liquid-liquid extraction procedure, atenolol was extracted from alkalinized body fluids with 5 ml chloroform-2-propanol (4:1, v/v). In the solid-phase extraction procedure, atenolol was isolated from plasma using a C8 column and methanol. Both extraction procedures were efficient in recovering atenolol and removing endogenous interferents. The RSDs and deviation from nominal values were lower than 10% for both within-day and between-day assays. The results show that there were no statistically significant differences in between-day variation. The t test showed that there were no significant differences between the real concentrations and the determined concentrations. The limit of quantitation was 10 ng/ml and the linear range was 10-5,000 ng/ml for both methods. These methods can be used in pharmacokinetic studies. PMID- 11863281 TI - Determination of thymol in human plasma by automated headspace solid-phase microextraction-gas chromatographic analysis. AB - A reliable and sensitive method was developed for determination of thymol in human plasma by automated headspace solid-phase microextraction (SPME). After enzymatic cleavage of thymol sulfate thymol was extracted by a 65 microm polydimethylsiloxane-divinylbenzene crimped fiber (Supelco) after addition of sodium chloride and phosphoric acid (85%). Desorption of the fiber was performed in the injection port of a gas chromatograph at 220 degrees C (HP 5890; 50 m x 0.2 mm I.D., 0.2 microm HP Innowax capillary column; flame ionization detection). Fibers were used repeatedly up to 40 analysis. The recovery was 5% after 35 min of extraction. The calibration curve was linear in the range of 8.1-203.5 ng ml( 1) with a limit of quantitation (LOQ) of 8.1 ng ml(-1). The within-day and between-day precision and accuracy were < or = 20% at the LOQ and <15% at higher concentrations according to international guidelines for validation of bioanalytical methods. After administration of a thymol-containing herbal extract only thymol sulfate, no free thymol, could be detected in human plasma, thus analysis of thymol was after enzymatic cleavage of thymol sulfate. It is concluded that the newly developed automated method can be used in clinical trials on bioavailability and pharmacokinetics of thymol-containing herbal medicinal products. PMID- 11863282 TI - Determination of alendronate in human urine as 9-fluorenylmethyl derivative by high-performance liquid chromatography. AB - A high-performance liquid chromatographic method for the quantitation of alendronate as the 9-fluorenylmethyl derivative (FMOC) in human urine is presented. The sample preparation involved coprecipitation with calcium phosphate, separation on diethylamine (DEA) solid-phase extraction (SPE) cartridge and derivatization with 9-fluorenylmethyl chloroformate in citrate buffer pH 11.9. Liquid chromatography was performed on an octadecylsilica column (150 x 4.6 mm, 3 microm particles); a gradient method with starting mobile phase acetonitrile-methanol-citrate/pyrophosphate buffer (20:15:65 v/v) was employed. The total run time was 21 min. The fluorimetric detector was operated at the following wavelengths: 260 nm (excitation) and 310 nm (emission). Pamdronate was used as the internal standard. The limit of quantitation was 3.5 ng/ml using 5 ml of urine. Within-day and between-day precision expressed by relative standard deviation was less than 8% and inaccuracy did not exceed 9%. The assay was applied to the analysis of samples from a pharmacokinetic study. PMID- 11863283 TI - High-performance liquid chromatographic-tandem mass spectrometric evaluation and determination of stable isotope labeled analogs of rofecoxib in human plasma samples from oral bioavailability studies. AB - A method for the simultaneous determination of a cyclooxygenase-2 inhibitor, 4-(4 methanesulfonylphenyl)-3-phenyl-5H-furan-2-one (rofecoxib, I) and [13C7]rofecoxib, (II), in human plasma has been developed to support the clinical oral bioavailability (BA) study of I. The method is based on high-performance liquid chromatography (HPLC) with atmospheric pressure chemical ionization tandem mass spectrometric (APCI-MS-MS) detection in the negative ionization mode using a heated nebulizer interface. Two different stable isotope labeled analogs of I were initially evaluated for their use as intravenous (i.v.) markers in the BA study. [13CD3]Rofecoxib was shown to be isotopically unstable in plasma and water containing solvent and an efficient deuterium exchange prevented its use in the study. On the other hand, the isotopic integrity of the subsequently synthesized [13C7]rofecoxib (II) was maintained, as expected, in plasma and other solvent systems. The results of these experiments clearly demonstrated the need for the careful evaluation of the isotopic integrity of the stable isotope labeled compound for the successful utilization of these compounds in BA studies and also as internal standards in the quantitative analysis of drugs in biological fluids. After liquid-liquid extraction of I, II, and internal standard (III) from plasma, the analytes were chromatographed on a narrow bore (100 mm x 3.0 mm) C18 analytical column, with mobile phase consisting of acetonitrile-water (1:1, v/v) at a flow-rate of 0.5 ml/min. The MS-MS detection was performed on a PE Sciex API III Plus tandem mass spectrometer operated in the selected reaction monitoring mode. The precursor-->product ion combinations of m/z 313-->257, 320-->292, and 327-->271 were used to quantify I, II, and III, respectively. The assay was validated in the concentration range of 0.1 to 100 ng/ml of plasma for both I and II. The precision of the assay (expressed as relative standard deviation) was less than 10% at all concentrations within the standard curve range, with adequate assay accuracy. The assay was utilized to support the clinical BA study in which oral doses of I were administered together with an i.v. dose of II to determine the oral BA of rofecoxib at 12.5- and 25-mg doses. PMID- 11863284 TI - Detection of fenspiride and identification of in vivo metabolites in horse body fluids by capillary gas chromatography-mass spectrometry: administration, biotransformation and urinary excretion after a single oral dose. AB - Studies related to the in vivo biotransforrmation and urinary excretion of fenspiride hydrochloride in the horse are described. After oral administration, the drug is metabolised by both phase I functionalisation and phase II conjugation pathways. Following enzymatic deconjugation, fenspiride and its phase I metabolites were isolated from post-administration biofluids using bonded co polymeric mixed mode solid-phase extraction cartridges to isolate the basic compounds. Following trimethylsilylation (TMS), the parent drug and metabolites were identified by capillary gas chromatography-mass spectrometry (GC-MS). Fenspiride (A) and seven metabolites (B-->G) arising from oxidation on both the aromatic and heterocyclic substructures were detected in urine. The positive ion electron ionisation mass spectra of the TMS derivatives of fenspiride and its metabolites provided useful information on its metabolism. Positive ion methane chemical ionisation-GC-MS of the derivatives provided both derivatised molecular mass and structural information. Unchanged fenspiride can be detected in post administration plasma and urine samples for up to 24 h. Maximum urinary levels of 100-200 ng ml(-1) were observed between 3 and 5 h after administration. After enzymatic deconjugation, the major phenolic metabolite (G) can be detected in urine for up to 72 h. This metabolite is the analyte of choice in the GC-MS screening of post-race equine urine samples for detection of fenspiride use. However, a distinct difference was observed in the urinary excretion of this metabolite between the thoroughbred horses used in UK study and the quarterbred and standardbred horses used for the USA administrations. PMID- 11863285 TI - Sensitive and rapid method for the determination of thalidomide in human plasma and semen using solid-phase extraction and liquid chromatography-tandem mass spectrometry. AB - Liquid chromatography-tandem mass spectrometric assays were developed for the sensitive, rapid and high throughput bioanalyses of thalidomide in human plasma and semen. The matrices were first stabilized with 0.025 M Sorensen's citrate buffer at pH 1.5 to prevent spontaneous hydrolysis. Buffered thalidomide was stable when stored at room temperature for 24 h and for up to three freeze-thaw cycles. Samples were extracted using SPE cartridges. Extracts were then injected into the LC-MS-MS equipped with a reversed-phase column and an APCI interface in the negative ion mode. Calibration curves for both matrices were linear with r>0.99 from 2 to 250 ng/ml and ng/g. Inter-assay precision (RSD) of plasma and semen calibration standards were 2.6-11.6 and 1.9-12.4%, respectively. Recoveries from plasma and semen were greater than 69 and 78%, respectively. Batch sizes of 100 samples per matrix were analyzed with a total run time of 5 h. The methods successfully determined concentrations of thalidomide from a clinical study to levels as low as 7 ng/ml plasma and 8 ng/g semen, respectively. PMID- 11863286 TI - Simultaneous determination of cloricromene and its active metabolite in rabbit aqueous humor by high-performance liquid chromatography. AB - A rapid and simple method was developed for the simultaneous separation and quantification of cloricromene, a coumarine derivative, and its active metabolite, cloricromene acid, in rabbit aqueous humor. The analyses were performed by high-performance liquid chromatography using a C18 reversed-phase column (Hypersil ODS) with UV detection at 318 nm. The mobile phase consisted of acetonitrile-water containing 1% triethylamine pH 3.5, adjusted with orthophosphoric acid. An acetonitrile gradient was necessary to achieve good separation within 13 min. Timolol was found to be a suitable internal standard. The retention times ranged from 5.72 to 11.25 min. A simple pre-treatment with acetonitrile containing 0.6% HCIO4 was used to deproteinize aqueous humor samples. The limit of quantitation ranged between 10 and 20 ng/ml. The recovery was >90%. The relationship between peak areas and concentration was linear over the range between 0.01 and 3.8 microg/ml, with r2 > 0.99. The assay provided good reproducibility and accuracy for both analytes and proved to be suitable for pharmacokinetic studies of cloricromene. PMID- 11863287 TI - High-performance liquid chromatographic assay for the simultaneous determination of sulfadoxine and pyrimethamine from whole blood dried onto filter paper. AB - A method using solid-phase extraction and high-performance liquid chromatography is evaluated for the simultaneous determination of sulfadoxine and pyrimethamine from 0.1 ml of whole blood dried onto filter paper. Extraction recoveries are about 60% for both drugs. The coefficient of variation for intra-assay precision, inter-assay precision and accuracy is less than 10% for sulfadoxine (10-100 microg/ml) and pyrimethamine (1-10 microg/ml). PMID- 11863289 TI - Rapid screening of urinary proline-hydroxyproline dipeptide in bone turnover studies. AB - In a recent report [J. Chromatogr. B 678 (1996) 165] a urinary hydroxyproline containing peptide has been preliminarily suggested as a possible alternative to hydroxyproline (HP) determination in bone resorption studies. For this purpose a simple and practical procedure was developed for a rapid high-performance liquid chromatographic (HPLC) assay of the peptide in non-hydrolyzed urine samples. Hundreds of randomly selected urine samples were assayed for both the peptide and HP, the latter in hydrolyzed urine, and a high correlation between them was found. The promising results prompted us to search for the postulated biomarker of bone resorption in urine samples of postmenopausal women examined as osteoporosis suspects. As an alternative to the HPLC determination, an equally rapid procedure has been developed for the peptide assay using capillary gas chromatography (GC) with flame ionization detection (FID). By means of a solid phase and a liquid-liquid phase extraction, involving ethyl chloroformate (ECF) as the derivatizing agent, two dipeptides and some urinary amino acids could be analyzed within 5 min. A high correlation between both HPLC and GC peptide assay was confirmed (r=0.944) and the compound was identified as proline-hydroxyproline (PHP) dipeptide. PMID- 11863288 TI - Determination of olanzapine in human blood by liquid chromatography-tandem mass spectrometry. AB - A liquid chromatographic-tandem mass spectrometric (LC-MS-MS) assay was developed and validated to quantitatively determine olanzapine (OLZ) concentrations in human blood. Liquid-liquid extraction, using n-butanol:cyclohexane (3:47, v/v), was used to isolate OLZ and its internal standard, LY170158, from the biological matrix. Chromatographic resolution of OLZ from endogenous interferences and known metabolites was accomplished with a MetaChem Monochrom HPLC column (4.6 x 150 mm, d(p) 5 microm). Detection occurred using a Perkin-Elmer Sciex API III Plus triple quadrupole mass spectrometer using positive ion APCI and multiple reaction monitoring (MRM). The linear dynamic range was from 5 to 500 ng ml(-1) based on a 0.25-ml aliquot of human blood. The inter-day precision (%RSD) and accuracy (%RE) ranged from 3.65 to 10.64 and from -2.14 to 3.07, respectively. Modifications to an existing assay for the determination of OLZ in human plasma were necessary. A different structural analog was used as the internal standard due to instability observed for the original analog when using human blood as the matrix. A second modification was the addition of the anti-oxidant sodium ascorbate to inhibit degradation of OLZ in human blood, as has been noted by other investigators. Upon fortification of human blood with sodium ascorbate (final concentration, 0.33 mM), OLZ was found to be stable for at least 1 week at -70 degrees C as well as through two freeze-thaw cycles. This assay, which will be used to investigate the distribution of OLZ in human blood, grants insight into the proper sample handling conditions needed to perform valid determinations of OLZ in human blood. PMID- 11863290 TI - Determination of urinary thiocyanate and nitrate using fast ion-interaction chromatography. AB - A simple and rapid chromatographic method for determination of nitrite, nitrate and thiocyanate is reported, and applied to the analysis of non-, medium and heavy smokers' urine samples. Ion-interaction liquid chromatography was carried out on a short 30 mm x 4.6 mm C18 column (3 microm particle size) with a mobile phase of 10 mM tetrabutylammonium phosphate in 20% MeOH. The chromatography was performed at an elevated temperature of 45 degrees C, at a flow-rate of 1 ml/min. Detection was by direct UV absorption at 230 nm. Sample preparation involved centrifugation and dilution, followed by sample clean-up on a C18 solid-phase extraction cartridge. The developed method proved both precise (% RSD <2%) and sensitive (standard detection limits <0.1 mg/l), and yielded total run times of under 10 min when applied to urine analysis of smokers and non-smokers, with thiocyanate eluting in under 5 min. PMID- 11863291 TI - High-throughput screening of potential inhibitors for the metabolism of the investigational anti-cancer drug 5,6-dimethylxanthenone-4-acetic acid. AB - By screening potential inhibitors of drug metabolism using the in vitro models, potential drug-drug interactions in vivo may be predicted with the use of appropriate pharmacokinetic principles. This study aimed to develop a rapid screening system using human liver microsomes to efficiently identify the potential inhibitors of DMXAA metabolism. Initial IC50 was estimated by using a two-point method, and then Ki values were determined if required and compared with those initial IC50 values. More than 100 compounds including known substrates and inhibitors of human uridine diphosphate glucuronosyltransferases (UGTs) and cytochrome P450 (CYP), anti-cancer drugs and xanthenone analogues were screened for their inhibitory effect on DMXAA glucuronidation and 6 methylhydroxylation in human liver microsomes. Both metabolites of DMXAA, DMXAA acyl glucuronide (DMXAA-G) and 6-hydroxymethyl-5-methylxanthenone-4-acetic acid (6-OH-MXAA), formed in human liver microsomes were quantitated by validated HPLC methods. The results indicated that there was a significant relationship (r2 = 0.966, P < 0.001) between the two-point IC50 values and the apparent Ki values for 20 compounds showing significant inhibitory effects on DMXAA metabolism, suggesting the usefulness of the two-point determination for the initial screening of compounds. This study has been completed using a strategy for rapid HPLC analysis and thus provided early access to detailed information for potential inhibitors of DMXAA metabolism and allows for further DMXAA-drug interaction studies. PMID- 11863292 TI - Identification of the major metabolite of 2,5-bis(5-hydroxymethyl-2-thienyl)furan (NSC 652287), an antitumor agent, in the S9 subcellular fraction of dog liver cells. AB - Alpha-Terthienyl (1) is a trithiophene found widely distributed in plants. Other naturally occurring trithiophenes are less widely distributed, but nonetheless exhibit potent antiviral and cytotoxic activities. A synthetic analog of 1, 2,5 bis(5-hydroxymethyl-2-thienyl)furan (2; NSC 652287) has recently been shown to possess exceptional activity and selectivity against several cell lines of the National Cancer Institute (NCI) anticancer drug screen. When incubated with the S9 subcellular fraction of dog liver cells, the concentration of 2 was observed to decline as a function of time, with a concomitant increase in a significant, time-dependent concentration of an unknown entity. The results of electron ionization mass spectrometric analysis of the metabolite indicate an increase in 14 amu over that of 2, leading to suspicions that either an oxidation or a methylation had occurred. Results of differential derivatization and accurate mass analysis allow us to propose that metabolism of 2 involves the biotransformation of one of the two hydroxymethyl groups of 2 into a carboxylic acid functionality. This is further supported by separate experiments involving chemical oxidation and S9 incubation of 5-[5-[5-hydroxymethyl-2-thienyl]-2 furanyl]-2-thiophenecarboxaldehyde: comparing the mass spectra and gas chromatographic retention times of the resulting products to those of the identified metabolite of 2 show all to be the same. PMID- 11863293 TI - Fast and sensitive capillary electrophoresis method to quantitatively monitor ibuprofen enantiomers released from polymeric drug delivery systems. AB - In this work, the capability of two polymeric drug delivery systems (DDS) containing racemic ibuprofen (IBU) for controlled release of IBU in different media was studied carrying out assays in-vitro. To quantitatively monitor the release of R(-)- and S(+)-IBU, a fast, sensitive and inexpensive capillary electrophoresis (CE) method was developed. To do this, different chiral selectors, temperatures, buffer compositions and pHs were tested. This new CE method uses bare silica columns together with a buffer containing 6% Dextrin in a 150 mM sodium tetraborate buffer at pH 9. Baseline separations of R(-)- and S(+) IBU were achieved in less than 5 min at 20 degrees C. By using this method, both enantiomers can be determined at concentrations as low as 1 microg/ml, allowing the detection of enantiomeric percentages of 0.5% of R(-)-IBU in the presence of 99.5% of the optical antipode. Moreover, the method shows a high reproducibility for the same day and different days. The usefulness of this method to quantitatively monitor the release of R(-)- and S(+)-IBU from two different polymeric DDS is demonstrated. It is shown that the release rate of IBU depends on the spacer of the side residue used in the polymeric device. Also, it is demonstrated that the release of both enantiomers is enzymatically activated in rat plasma. PMID- 11863294 TI - Separation of human serum transferrins with different iron-binding states by high performance liquid chromatography using a pyridinium polymer column. AB - Four molecular forms of transferrins with different iron-binding states were separated by HPLC using a pyridinium polymer column. The elution order was monoferric transferrin bound to the C-site, holotransferrin, apotransferrin and monoferric transferrin bound to the N-site. Human sera were also analyzed with the column, and ICP-MS combined with HPLC was used to detect iron in each peak. Transferrin peaks separated by HPLC were also confirmed by an immunological method. The percentages of iron saturation in transferrins obtained by the HPLC method were compared with the values calculated from clinical data. PMID- 11863295 TI - Direct analysis of indomethacin in rat plasma using a column-switching high performance liquid chromatographic system. AB - We have established a robust, fully automated analytical method for the determination of indomethacin in rat plasma using a column-switching high performance chromatographic system. The system consists of a precolumn and an analytical column connected in series via a switching valve. When a 50-microl portion of rat plasma containing a therapeutic level of indomethacin was applied directly to the system, the drug was automatically enriched in the precolumn (TSK BSA-ODS) by on-line solid-phase extraction. After elution of the plasma proteins, the analyte was automatically transferred to the analytical column (Zorbax Eclipse XDBC18) where chromatography was performed using isocratic elution and UV absorption detection at a wavelength of 254 nm. The separation mobile phase consisted of methanol-0.1% phosphoric acid (70:30, v/v) at a flow-rate of 1 ml/min. The calibration line for indomethacin showed good linearity in the range 50-10 000 ng/ml (r>0.999) with the detection quantification of 50 ng/ml (RSD=2.6%). Accuracy ranged from -0.62 to 3.22%, and the within- and between-day precision of the assay was better than 6% across the calibration range. The analytical sensitivity and accuracy of this assay is suitable for characterization of the pharmacokinetics of topical administration of imdomethacin to rats. The method has been successfully used to provide pharmacokinetic data in a large number of diverse pharmaceutical studies. PMID- 11863296 TI - High-performance liquid chromatography with spectrophotometric and electrochemical detection of a series of manganese(III) cationic porphyrins. AB - Recent studies have revealed potent pharmacological activities of manganese containing cationic porphyrins. An analytical method employing high-performance liquid chromatography with spectrophotometric and electrochemical detection (HPLC UV/EC) suitable for in vivo applications is described for a series of manganese(III) cationic porphyrins with good separation and resolution. In particular, this method resolved the four atropisomers of manganese(III) meso tetrakis(N-ethylpyridinium-2-yl)porphyrin (MnTE-2-PyP5+ or AEOL-10113), verified by mass spectrometry. Electrochemical and spectrophotometric methods of detection were compared using manganese(III) meso-tetrakis(1,3-diethylimidazolium-2 yl)porphyrin (MnTDE-2-ImP5+ or AEOL-10150), the lead catalytic antioxidant of this series. Both methods of detection were quantitative, but electrochemical detection, although less specific for in vivo applications, appears to be considerably more sensitive than spectrophotometric detection. PMID- 11863297 TI - Rapid determination of nevirapine in human plasma by gas chromatography. AB - A sensitive and rapid gas chromatographic method has been developed to determine the levels of the HIV-1 non-nucleoside reverse transcriptase inhibitor nevirapine in human plasma. Quantitative recovery following liquid-liquid-extraction with diethylether from 500 microl of human plasma was achieved. Subsequently, the assay was performed with a CP-Sil 5CB capillary column, 15 m x 0.32 mm x 1.0 microm film thickness with a nitrogen-phosphorous-detector (NPD), Helium 5.0 was used as carrier gas with a constant inlet pressure of 7 p.s.i. Linear standard curves were obtained for concentrations ranging from 10 to 20 000 ng/ml. The calculated intra- and inter-day coefficients of variation were below 8%. PMID- 11863298 TI - Quantitation of tamsulosin in human plasma by liquid chromatography-electrospray ionization mass spectrometry. AB - A highly sensitive method for quantitation of tamsulosin in human plasma using 1 (2,6-dimethyl-3-hydroxylphenoxy)-2-(3,4-methoxyphenylethylamino)-propane hydrochloride as the internal standard (I.S.) was established using liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS). After alkalization with saturated sodium bicarbonate, plasma were extracted by ethyl acetate and separated by HPLC on a C18 reversed-phase column using a mobile phase of methanol-water-acetic acid-triethylamine (620:380:1.5:1.5, v/v). Analytes were quantitated using positive electrospray ionization in a quadrupole spectrometer. LC-ESI-MS was performed in the selected ion monitoring (SIM) mode using target ions at m/z 228 for tamsulosin and m/z 222 for the I.S. Calibration curves, which were linear over the range 0.2-30 ng/ml, were analyzed contemporaneously with each batch of samples, along with low (0.5 ng/ml), medium (3 ng/ml) and high (30 ng/ml) quality control samples. The intra- and inter-assay variability ranged from 2.14 to 8.87% for the low, medium and high quality control samples. The extraction recovery of tamsulosin from plasma was in the range of 84.2-94.5%. The method has been used successfully to study tamsulosin pharmacokinetics in adult humans. PMID- 11863299 TI - Development of a sensitive high-performance thin-layer chromatography method for estimation of ranitidine in urine and its application for bioequivalence decision for ranitidine tablet formulations. AB - A sensitive and simple HPTLC method was developed for estimation of ranitidine in human urine. The drug was extracted from urine after basification using dichloromethane. Dichloromethane extract was spotted on silica gel 60 F254 TLC plate and was developed in a mixture of ethyl acetate-methanol-ammonia (35:10:5 v/v) as the mobile phase and scanned at 320 nm. The RF value obtained for the drug was 0.67 +/- 0.03. The method was validated in terms of linearity (50-400 ng/spot), precision and accuracy. The average recovery of ranitidine from urine was 89.35%. The proposed method was applied to evaluate bioequivalence of two marketed ranitidine tablet formulations (150 mg, Formulation I and Formulation 2) using a crossover design by comparing urinary excretion data for unchanged ranitidine in six healthy volunteers. Various pharmacokinetic parameters like peak excretion rate [(dAU/dt)max], time for peak excretion rate (tmax), AUC0-24, AUC0-infinity, cumulative amount excreted were calculated for both formulations and subjected to statistical analysis. The relative bioavailability of Formulation 2 with respect to Formulation 1 was 93.76 and 95.31% on the basis of AUC0-24 and cumulative amount excreted, respectively. Statistical comparison of various pharmacokinetic parameters indicated that the two ranitidine tablet formulations are bioequivalent. PMID- 11863300 TI - Determination of alachlor and its metabolites in rat plasma and urine by liquid chromatography-electrospray ionization mass spectrometry. AB - A method based on liquid chromatography (LC) in combination with mass spectrometry (MS) for the analysis of alachlor (ALA) and its metabolites, 2 chloro-N-[2,6-diethylphenyl]acetamide (CDEPA) and 2,6-diethylaniline (DEA), in rat plasma and urine has been developed. 13C-labeled ALA was used as the internal standard for quantitation. The analyte in plasma or urine was isolated using a Waters Oasis HLB extraction plate. The mass spectrometer was operated in the ESI MS-SIM mode with a programming procedure. The retention times for ALA, CDEPA and DEA were 1.84, 3.11 and 4.12 min, respectively. The limits of quantification (LOQ) for ALA, CDEPA and DEA were 2.3, 0.8 and 0.8 ng per injection, respectively. The linear fit of analyte to mass response had an R2 of 0.99. Reproducibility of the sample handling and LC-MS analysis had a RSD of < or = 10%. The average recoveries for these analytes in rat plasma were better than 90%. Similar results were obtained with rat urine. PMID- 11863302 TI - Effusive constrictive pericarditis due to rheumatoid arthritis revealed by pericardiocentesis with simultaneous pressure recording--a case report. AB - A 59-year-old man had a history of rheumatoid arthritis. He presented with incurable pericardial effusion. He was repeatedly treated with pericardiocentesis with only transient attenuation of his symptoms because the underlying pericardial constriction had been overlooked. This time the authors diagnosed effusive constrictive pericarditis due to rheumatoid arthritis using the hemodynamic findings observed before and after pericardiocentesis. PMID- 11863301 TI - Venous thrombosis from air travel: the LONFLIT3 study--prevention with aspirin vs low-molecular-weight heparin (LMWH) in high-risk subjects: a randomized trial. AB - The LONFLIT1 and 2 studies established that in high-risk subjects after long (>10 hours) flights, the incidence of deep venous thrombosis (DVT) may be between 4% and 6%, The LONFLIT3 study aimed to evaluate methods of prevention in high-risk subjects. Of 467 subjects contacted for the study, 300 were included. These 300 subjects at high risk for DVT were randomized, after informed consent, into three groups: 1) a control group that had no prophylaxis; 2) an aspirin treatment group, in which patients were treated with 400 mg (tablets of oral, soluble aspirin; one dose daily for 3 days, starting 12 hours before the beginning of the flight); and 3) a low-molecular-weight heparin (LMWH) group, in which one dose of enoxaparine was injected between 2 and 4 hours before the flight. The dose was weight-adjusted (1,000 IU [equivalent to 0.1 mL per 10 kg of body weight). Subjects with potential problems due to prophylaxis with aspirin or LMWH or at risk of drug interactions were excluded. Of the 100 included subjects in each group, a total of 249 subjects completed the study (dropouts due to low compliance or traveling/connections problems were 17%). Age and sex distribution were comparable in the three groups as well as risk distributions. Mean age was 47 (range, 28-75; SD, 11; 65% males). Of the 82 subjects in the control group, there were 4.82% of subjects with DVT with two superficial thromboses. In total 4.8% of limbs suffered a thrombotic event. Of 84 subjects in the aspirin treatment group, there were 3.6% of patients with DVT and three superficial thrombosis. In total 3.6% of limbs had a thrombotic event. In the LMWH group (82 subjects), there were no cases of DVT. One superficial thrombosis was documented. In total only 0.6% of limbs had a thrombotic event (p<0.002 in comparison with the other two groups). DVT was asymptomatic in 60% of subjects; 85% of DVTs were observed in passengers in non-aisle seats. Mild gastrointestinal symptoms were reported in 13% of patients taking aspirin. One dose of LMWH is an important option to consider in high-risk subjects during long-haul flights. PMID- 11863303 TI - Cardiomyopathy and atrioventricular block in Emery-Dreifuss muscular dystrophy--a case report. AB - A 32-year-old woman is described as having the following characteristics of Emery Dreifuss muscular dystrophy: humeroperoneal muscular atrophy and weakness, neck and elbow contractures with sinus bradycardia, first-degree atrioventricular block, and dilated cardiomyopathy. The biopsy specimen of skeletal muscle showed dystrophic character; a cardiac endomyocardial biopsy specimen showed adipose tissue infiltration and deposition of antihuman IgG. Emery-Dreifuss muscular dystrophy is an X-linked recessive myopathy. The patient had no familial background of the disease. This patient might have a sporadic inheritance pattern with severe cardiac involvement. PMID- 11863304 TI - Intravenous immunoglobulin-related acute coronary syndrome and coronary angiography in idiopathic thrombocytopenic purpura--a case report and literature review. AB - In acute coronary syndromes, GPIIb/IIIa platelet inhibitors have demonstrated a reduction in recurrent myocardial ischemia. Conversely, one might expect that enhancing platelet activity in patients in acute coronary syndromes would have the opposite effect. We report a patient with idiopathic thrombocytopenic purpura (ITP) that had recurrent myocardial ischemia associated with administration of intravenous immunogloblin (IVIG). Literature is reviewed. PMID- 11863305 TI - Cardiovascular risk factors do not predict clinically defined restenosis after percutaneous transluminal angioplasty for lower limb ischemia. AB - The purpose of this study was to evaluate risk factors predicting restenosis and primary patency after percutaneous transluminal angioplasty. Follow-up data (including cardiovascular risk factor scores according to SCVIR criteria, preinterventional and postinterventional clinical data and patient history) of all patients who underwent successful percutaneous transluminal angioplasty for lower limb ischemia were analyzed retrospectively and patients, relatives, or referring physicians underwent a telephone interview. Patients with incomplete follow-up data were examined by means of a clinical examination, including Doppler measurements and treadmill test. Additionally all angiograms were evaluated to calculate lesion length, number of treated lesions, lesion type (SCVIR score), and runoff. The outcome was categorized into four groups: early recurrence (< 1 month, group I), mean recurrence (1-6 months, group II), late recurrence (>6 months, group III), and no recurrence (group IV). According to common concepts group I was defined as early (thrombotic) reocclusion, group II as clinically defined restenosis, and group III as progression of atherosclerosis. One hundred thirty-seven patients underwent percutaneous transluminal angioplasty of 148 extremities. The groups differ significantly only with respect to a higher diabetes score for group I in comparison to group IV (p=0.002, Kruskal-Wallis test), and a worse runoff of group I compared with group IV (p =0.008). There was a trend toward a higher diabetes score for group II in comparison to group IV (p = 0.014). There were no differences with regard to hyperlipemia, hypertension, and tobacco use between patient groups. Mean primary patency was 436 days. Predictors for lower patency rates were diabetes mellitus (p<0.001), runoff (p=0.005), and number of treated lesions (p=0.007) in a stepwise, multiple regression analysis. Patients with clinically defined restenosis showed no specific risk factor profile in this study. Predictors for lower primary patency were diabetes mellitus, number of treated lesions, and runoff. PMID- 11863306 TI - Validity of the Leg-O-Meter, an instrument to measure leg circumference. AB - The objective of this study was to validate the Leg-O-Meter measure against the clinical assessment of edema made by physicians using data from a 1-year follow up study of unselected patients with chronic venous disease of the leg (CVDL). The Leg-O-Meter consists of a tape measure fixed to a stand attached to a small board on which the patient is in standing position. Its reliability has been shown to be above 97%. Data from the Venous Insufficiency Epidemiologic and Economic Study (VEINES) were used: 1,521 patients from France, Belgium, Italy, and Quebec (Canada) who spontaneously consulted a physician between 1994 and 1995 with a complaint resulting from venous problems of the legs were included. Baseline variables included leg circumference measurements using the Leg-O-Meter; physicians were also asked to diagnose edema and report it as present or absent on each leg. Clinical edema and leg circumferences were assessed again 3 to 6 months after the baseline visit and 12 months after baseline. The tape measure of the Leg-O-Meter was fixed at 13 cm from the floor. The first and last assessments were used to evaluate the variation in edema during the follow-up period. Clinical variation in edema status was assessed as follows: improved, if edema was diagnosed at baseline but not at the final visit; unchanged, if edema was diagnosed at both visits; and worsened, if there was no diagnosis of edema at baseline but a diagnosis of edema was made at the final visit. Variation in measured edema was classified as improved if there was a decrease in leg circumference of more than 1 cm between baseline and final evaluation; unchanged, if the difference in leg circumference was between plus or minus 1 cm between the 2 assessments; and worsened, if there was an increase in leg circumference greater than 1 cm between the 2 assessments. Data-driven cut-off points were also used: 1.3 cm and 1.5 cm. Sensitivity and specificity of the Leg-O-Meter using physician diagnosis as "gold standard" were calculated. In addition, receiver operating characteristics (ROC) curves were calculated by using the 3 different leg circumference cut-off points in order to determine the accuracy of the Leg-O Meter to detect changes in edema. The overall accuracy of the Leg-O-Meter was 0.84 (standard error (se) = 0.06). Accuracy was greater when 1.5 cm was used as a cut-point. The Leg-O-Meter is an objective, reliable, and standardized instrument to assess patients over time. A change of 1.5 cm between 2 measurements gives a valid estimate of improvement or worsening of edema, when compared to physicians' diagnosis. The Leg-O-Meter is also sensitive to any changes in leg circumferences, which is an advantage over the clinical evaluation of edema. PMID- 11863307 TI - Occlusive thromboaortopathy (Takayasu disease): clinical and angiographic features and a brief review of literature. AB - The aim of this study was to evaluate the diagnostic criteria and angiographic classifications of Takayasu arteritis by presenting the clinical, angiographic, and prognostic findings and a prospective follow-up of 78 patients. Occlusive thromboaortopathy or Takayasu arteritis is a large vessel vasculitis. The disease is systemic with an autoimmune and genetic etiology. The complete clinical and angiographic manifestations are reported for 78 cases based on diagnostic criteria of the American College of Rheumatology with a mean 6 +/- 3.2 years follow-up. The mean age was 34.7 and female:male ratio was 3.6:1. According to National Institute of Health criteria, 61.5% of patients were in the acute phase of disease with systemic symptoms such as fever, weight loss, malaise, and elevated C-reactive protein levels. Immunologic markers, such as antinuclear antibody and antineutrophil cytoplasmic antibodies, were negative. The tuberculin test result was positive in 47%. Vascular bruit was present in 89%. Almost all patients had stenoses, occlusions, or aneurysmatic changes of the aorta and its main branches. Hypertension was detected in 58% and left ventricular hypertrophy was initially present in 22 (28%) patients. The angiographic manifestations were classified as type I, cervicobrachial type with 20 cases (25.6%); type II, thoracoabdominal type with 13 cases (16.6%); type III, peripheral type with 10 cases (12.8%); and type IV, generalized type with 35 cases (44.8%). The coronary arteries were involved in 6 cases, pulmonary arteries in 11 initially 5 in follow up (16 cases), and renal arteries in 28 cases, respectively. A good correlation of the clinical manifestations and the prognosis was observed. During follow-up, five patients suffered from myocardial infarction, six had cerebrovascular accident, seven patients underwent aortic valve replacement, and six patients died (mortality rate, 7.6%). The specificity and sensitivity of diagnostic criteria were 94% and 76%, respectively. In contrast to ours and Nasu's classification in the new classification of Numano, some angiographic types and subtypes of Takayasu arteritis are not present in our patients. PMID- 11863308 TI - A new technique for the quantification of peripheral edema with application in both unilateral and bilateral cases. AB - Current noninvasive techniques for the routine and frequent quantification of peripheral lymphedema in patients are total limb volume measurement (by water immersion or by circumferential measurements) and bioelectrical impedance analysis (BIA). However both of these techniques require standardizing the measurement using a contralateral measurement from the unaffected limb. Hence these techniques are essentially restricted to unilateral lymphedema. This paper describes the results from a preliminary study to investigate an alternative approach to the analysis of the data from multiple frequency BIA to produce an index of lymphedema without the need for normalization to another body segment. Twenty patients receiving surgical treatment for breast cancer were monitored prior to surgery and again after diagnosis with unilateral lymphedema. The data recorded were total limb volume, by circumferential measurements; and BIA measurements of both limbs. From these measurements total limb volumes and extracellular fluid volumes were calculated and expressed as ratios of the affected limb to that of the unaffected limb. An index of the ratio of the extracellular fluid volume to the intracellular fluid volume was determined. This ECW/ICW index was calculated for both the affected and unaffected limbs at both measurement times. Results confirmed that the established techniques of total limb volume and extracellular fluid volume normalized to the unaffected contralateral limb were accurate in the detection of lymphedema (p < 10(-6)). Comparison of the ECW/ICW index from the affected limb after diagnosis with that from the pre-surgery measurement revealed a significant (p < 10(-6)) and considerable (75%) increase. The results of this pilot study suggest that by using multiple frequency bioelectrical impedance analysis, an index of the ECW/ICW ratio can be obtained and this index appears to have an equal, or better, sensitivity than the other techniques in detecting lymphedema. More importantly, this index does not require normalization to another body segment and can be used to detect all types of peripheral edema including both unilateral and bilateral lymphedema. PMID- 11863309 TI - Stenotrophomonas maltophilia endocarditis: a systematic review. AB - The disease characteristics, management, and outcome of Stenotrophomonas maltophilia endocarditis were evaluated by examining the reports on the subject identified through a comprehensive literature search. Twenty-three (17 male) cases of S.. maltophilia endocarditis were identified. Mean age was 41 +/- 15 years. All patients presented with fever. Prosthetic valves were involved in 12 (52%) cases. Among native valves, the aortic valve was most frequently involved (50%), followed by the tricuspid valve (36%). Twenty (87%) patients had underlying risk factors for the development of endocarditis, including prior valvular or congenital heart disease surgery (60%), intravenous drug abuse (32%), and infected intravascular lines (18%). The endocarditis was postoperative in 14 patients. Seventeen (74%) patients experienced complications including septic embolism (23%), cardiac abscesses (23%), and congestive heart failure (18%). A combination of two or more antibiotics was used in all cases except one. The frequently used antibiotics were aminoglycosides (59%), trimethoprim sulfamethoxazole (48%), and penicillins (48%). One half of the patients required cardiac surgery, but the proportion of surgically treated cases was higher among prosthetic valve endocarditis (62%). Mortality was 39% and was equally distributed between patients with prosthetic and native valve endocarditis. The S. maltophilia endocarditis carries high complication and mortality rates. The antibiotic regimen should consist of a combination of multiple antibiotics guided by the sensitivity panel. Early surgery may be considered in patients not responding to antibiotic treatment and in those with prosthetic valve endocarditis. PMID- 11863310 TI - Long-term effect of lipid-lowering therapy on atherosclerosis of abdominal aorta in patients with hypercholesterolemia: noninvasive evaluation by a new image analysis program. AB - Recent clinical studies have demonstrated that 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors are effective in the prevention of cardiovascular events and regression of atherosclerotic lesions evaluated by angiography. In this study, the authors investigated how lipid-lowering therapy effects on the progression of aortic atherosclerosis by using plain and enhanced computed tomography (CT) of the lower abdominal aorta. Twenty-nine hyperlipidemic patients (mean age 61.4 +/- 7.2 yr) were enrolled in a prospective open-labeled study. All patients underwent baseline CT scanning of abdominal aorta, screening for serum lipid profile and coagulation-fibrinolysis measurement, then treatment with simvastatin was begun. After 2 years, a follow-up CT scan was done and atherosclerotic lesions were compared between baseline and on-treatment scan. In spite of significant improvement of lipid and fibrinolytic profiles by simvastatin administration, mean aortic wall thickening volume (AWV) was increased during observation period. When patients were divided into subgroups by the levels of on-treatment LDL cholesterol (LDL-C), development rate of AWV was more potently suppressed in patients whose on-treatment LDL-C were below 125 mg/dL (median LDL-C). We could not find any associations of coagulation fibrinolysis measurements with atherosclerotic lesions. In regard to aortic calcification volume (ACV), low levels of total and HDL cholesterol and higher age were associated with aortic calcification at baseline. These results suggest that aggressive treatment with LDL-C below 125 mg/dL may suppress the progression of wall thickening and factors that promote arterial calcifications and those for wall thickening may be different. PMID- 11863311 TI - Angiography-guided routine coronary stent implantation results in suboptimal dilatation. AB - The resistance of the atherosclerotic lesion counteracts the expansion of the stent, resulting in suboptimal stent expansion. Intravascular ultrasound provides more precise information on stent expansion than coronary angiography but adds cost and time to the percutaneous transluminal coronary angiography procedure. The aim of this study was to evaluate the need for intravascular ultrasound at routine angiography-guided high pressure stent implantation by comparing stent expansion with predefined intracoronary ultrasound criteria for optimal stent implantation. In 32 patients, 48 stents (35 NIR, 12 AVE, and 1 Cordis) were implanted in A, B, and C stenoses using a high-pressure inflation technique until an optimal result was achieved according to angiography. Stent expansion was then evaluated by intravascular ultrasound as minimal lumen diameter, minimal lumen area, proximal and distal stent area, and a minimal lumen area symmetry index. These variables were then compared with the nominal stent size in vitro. Finally the stents were also evaluated with respect to the MUSIC criteria, ie, strict criteria regarding symmetry, apposition, and vessel geometry according to intravascular ultrasound after stent expansion. Forty-five stents could be completely analyzed. The mean balloon inflation pressure was 12.8 (range, 10-17) atm. The nominal stent size was not achieved in any patient. Minimal lumen diameter attained 77% and minimal lumen area 78% of expected nominal values (p<0.0001), distal stent area 88% (p < 0.001), and proximal stent area 92% (ns). Application of the MUSIC criteria showed that almost all stents (96%) had good stent apposition and symmetry index. Optimal proximal stent entrance was found in 70%. Optimal minimal lumen area in comparison to the reference areas was present in 41%. This lead to fulfilling of all MUSIC criteria in 47% of the stents. If nominal stent size had been achieved, symmetry index and apposition would have been fulfilled in all cases and optimal minimal lumen area increased to 75%. Acceptable proximal entrance however would have decreased to 55% and the fulfillment of all MUSIC criteria would increase only to 52%. In routine angiography-guided stent implantation in stenoses with a wide range of severities using modern stents and high pressure inflation technique to reach a visually optimal result, the nominal stent size was never achieved mainly due to residual intrastent stenosis. If nominal stent size had been achieved, the results would have improved only marginally and would still be suboptimal in almost half of the stents. These results highlight the shortcoming of angiography and the need for intravascular ultrasound in choosing correct stent size. PMID- 11863312 TI - Effects of beraprost sodium (Dorner) in patients with diabetes mellitus complicated by chronic arterial obstruction. AB - The effects of the prostaglandin I2 derivative beraprost sodium (Dorner) on ankle pressure index (AP; ankle joint-to-upper extremity systolic pressure ratio), subjective symptoms, and intermittent claudication were investigated in diabetic patients with arteriosclerosis obliterans (ASO). Forty patients (25 men and 15 women), mean age 63.9 years, were enrolled in this study. ASO was grade I in 30 patients, grade II in seven, grade III in one, and grade IV in two according to the Fontaine classification. They were administered six tablets (20 microg/tablet) of beraprost sodium daily for 6 months. At 3 and 6 months, API had significantly increased and symptoms such as coldness, numbness, and lack of feeling in the lower extremities were significantly improved. Ten evaluable patients increased ambulatory distance by approximately threefold, suggesting an improvement in intermittent claudication. Adverse reactions were experienced by five (12.5%) of the 40 patients (one case each of headache, dull headache, pain in the posterior region of the neck, heartburn, stomach discomfort, and anemia), but all were mild and resolved without treatment. Beraprost sodium was shown to improve API and symptoms in the lower extremities in diabetic patients with ASO, suggesting that it is useful in treating peripheral circulatory disorders in such patients. PMID- 11863313 TI - Evaluation of plasma endothelin-1 levels in patients with cerebral infarction. AB - Endothelin-1 (ET-1) is a vasoconstrictor peptide derived from endothelium. Many authors have shown that ischemic stroke is associated with elevated plasma ET-1 levels. Also, the present findings related to plasma ET-1 levels with clinical status, size of the infarction, location of the infarction, and prognosis of the cerebral infarction were contradictory. In this study, plasma ET-1 levels in 30 patients with cerebral infarction within 72 hours after the onset of focal neurologic deficit and at their seventh day postinfarction were measured by a microplate enzyme immunoassay. Thirty sex- and age-matched healthy subjects were accepted as a control group. The mean plasma ET-1 concentrations in patients on admission, in patients at day 7, and in control subjects were 1.93 +/- 1.79, 1.03 +/- 1.02, and 0.65 +/- 0.32 fmol/mL, respectively. The mean plasma ET-1 level of patients on admission was found to be significantly higher than in patients at day 7 and in control subjects (p < 0.05). No significant difference in ET-1 levels was observed between the patients at day 7 and control subjects. Furthermore, there was no correlation between plasma ET-1 concentration and size of infarction, location of infarction, degree of clinical neurologic deficit, or prognosis of cerebral infarction. It was concluded that plasma ET-1 levels shortly after ischemic stroke were increased, which may be associated with the acute-phase reaction of cerebral infarction and may have deleterious effects on development of neuronal injury. PMID- 11863314 TI - Spontaneous coronary artery dissection: the clinical spectrum. AB - Spontaneous coronary artery dissection is a rare but increasingly reported cause of myocardial ischemia and sudden death. It is more commonly seen in younger age groups and has a predilection for postpartum women. The clinical spectrum of its presentation includes angina, myocardial infarction, cardiogenic shock, and death. No specific cardiac risk factors have been associated with its occurrence. In postpartum patients, it is presumed that dissection results from pregnancy induced degeneration of collagen and the additional stresses of parturition. The sporadic nature of spontaneous coronary artery dissection has precluded the development of any consensus for a medical approach, and treatment is usually tailored to individual patient needs. A case of postpartum spiral dissection of the left anterior descending coronary artery with successful medical management is reported. PMID- 11863316 TI - Vascular diseases: new results with pentoxifylline. PMID- 11863315 TI - Endothelial functions in pathophysiology of thrombosis and fibrinolysis: late spontaneous recanalization of an occluded internal carotid artery--a case report. AB - The use of oral contraceptives is a potential cause of ischemic stroke in young women. The risk of stroke is higher when contraceptives contain high levels of estrogens. A thrombotic occlusion of the right internal carotid artery, seen on ultrasound, developed in a patient who was taking high-dose estrogen contraception. Recanalization occurred several months later by spontaneous thrombolysis and was confirmed by cerebral angiography. This case suggests that the activation of endothelial spontaneous antithrombotic mechanisms may allow the dissolution of a thrombus, once the cause of the thrombosis has been identified and removed and when the endothelium has maintained its functional integrity. PMID- 11863317 TI - Efficacy and safety of topical hirudin (Hirudex): a double-blind, placebo controlled study. AB - Hirudin, a thrombin inhibitor isolated from the leech Hirudo medicinalis, has been long known for its anticoagulant effects. In 1990, the former German BGA has published a monograph on Hirudo medicinalis extract (containing Hirudin and Eglin), stating that its local use is indicated in bruises with or without hematoma. The aim of this double-blind, placebo-controlled clinical trial was to evaluate the efficacy and safety of a Hirudin-containing cream (Hirudex cream; Hirudo medicinalis extract 280 UI/100 g) in patients affected with bruises with or without hematoma. 60 men and women between the ages of 18 and 65 years with a unilateral acute musculoskeletal injury (bruise) with or without hematoma were included. Dosage schedule and application route for both treatments were the following: 3-4 daily applications for 5 days for a total of 15-20 administrations during the whole study period. In the Hirudin group, a highly statistically and clinically significant improvement were noted. Although a statistical improvement was also seen in patients treated with placebo, this was less pronounced, and a highly significant between-group difference was noted for all three major efficacy parameters at each follow-up visit in favour of Hirudin. Both the patients and the investigator considered the overall assessment of efficacy at the end of the study to be significantly better (p < 0.001) in the Hirudin group than in the placebo group. Results of this study suggest that Hirudin is an effective local treatment in patients with mild to moderate bruises. PMID- 11863318 TI - Effectiveness of beclomethasone dipropionate aerosolized through different nebulizers to asthmatic patients. AB - The aim of our study was to verify if the type of nebulizer used could influence the results of aerosol therapy with beclomethasone dipropionate (BDP) in mild allergic asthma. We assigned 27 asthmatics allergic to grasses to 3 groups and treated them from May to July 1998 with aerosol therapy with BDP (800 microg) b. i. d. via nebulizer + pMDI salbutamol (200 microg) if necessary. Each group used a different type of nebulizer: jet nebulizer with glass ampoule (group JG); jet nebulizer with polycarbonate ampoule (group JP); ultrasonic nebulizer (group US). During the study patients underwent periodic lung function tests and methacholine bronchial challenges, recorded twice a day self-monitoring PEF and filled out a daily diary for the presence and intensity of asthmatic symptoms. At the end of the study the provocative dose of methacholine causing a 20% fall in FEV1 (PD20), the self-monitoring PEF and the clinical scores were all greatly improved, but without any statistically significant difference among the three groups. On the contrary, the variations during the study of basal spirometric parameters (specifically FEV1, PEF, FEF25) were significantly better in jet nebulizer groups than in group US. The results coming from the aerosol characterization that we carried out for each of the three nebulizers confirmed the clinical findings, since jet nebulizers showed greatly lower MMAD than the ultrasonic nebulizer (2.9 and 3.7 vs 5.8). Our data suggest that jet nebulizers are more appropriate than ultrasonic nebulizers for delivering BDP in aerosol therapy. PMID- 11863319 TI - The use of antileukotrienes in paediatrics. AB - Allergic diseases include a variety of different illnesses (rhinitis, conjunctivitis, asthma, urticaria, dermatitis) whose physiological and pathological basis is the release of chemical mediators such as histamine, PAF (platelet activating factor), metabolites of arachidonic acid and chemotactic factors from mastocytes, basophils and eosinophils. The numerous drugs used for allergy treatment now include the new pharmacological category of cysteinyl leukotriene antagonists. The cysteinyl leukotrienes (LTC4, LTD4 and LTE4) are chemical mediators of the inflammation involved in the pathogenesis of asthma, the biological effects of which are bronchial constriction and an increase in both mucus secretion and vascular permeability. Recent studies carried out above all on adult patients suggest that antileukotrienes can play an important part not only in the acute phase but also in controlling the chronic development of bronchial asthma. Antileukotrienes have also been successfully used by some authors to control atopic dermatitis and urticaria. Though further controlled testing will be required, these applications broaden the possible range of treatments for allergic disease in all its many aspects. PMID- 11863320 TI - Efficacy of a single-dose ondansetron for preventing post-operative nausea and vomiting after laparoscopic cholecystectomy with sevoflurane and remifentanil infusion anaesthesia. AB - In this randomized study we compared the efficacy of ondansetron 4 mg with ondansetron 8 mg for the prevention of postoperative nausea and vomiting (PONV) after laparoscopic cholecystectomy with sevoflurane and remifentanil infusion anaesthesia. Sixty patients were randomized to receive ondansetron 8 mg (30 pts) or ondansetron 4 mg (30 pts) before the induction of anaesthesia with thiopental and remifentanil. Anaesthesia was maintained with sevoflurane (0.5 MAC), oxygen and remifentanil infusion (0.25, 0.35, 0.5 microg/kg/min). Postoperative analgesia was provided by intravenous ketorolac 60 mg. The incidence of PONV, the pain score, and the analgesic requirement were recorded for 24 hours. There was no difference among groups in patient characteristics, risk factors for PONV, or side effects. During the first 6 h postoperatively, the incidence of PONV after ondansetron 4 mg and 8 mg were similar (p < 0.001). After 6 h the incidence of PONV increased significantly in patients who had received ondansetron 4 mg (p = 0.01) and was greater than that in patients who had received ondansetron 8 mg (p = 0.001). We conclude that single-dose ondansetron 8 mg is more effective than ondansetron 4 mg in the prevention of PONV after laparoscopic cholecystectomy. This surgery is associated with a high incidence of postoperative nausea and vomiting. A single dose of IV ondansetron 8 mg is well tolerated and decrease the number of nausea and vomiting episodes after surgery. PMID- 11863321 TI - Diastolic filling in hypertrophied hearts of elite runners: an echo-Doppler study. AB - The aim of this study was to establish if the physiologic adaptations following a prolonged physical training could influence the diastolic function in a professional Olympic male runner group. From February to December 1999 we studied 25 athletes (Group I) during the period of maximal training compared with 18 healthy sedentary subjects of matched age and sex (Group II). We used mono and bidimensional Echocardiography to assess left ventricular structure and systolic function. The diastolic function was evaluated by Doppler method assessing transmitral and venous pulmonary flow. From the comparison between the two groups, we found great differences in the interventricular septum and the posterior wall thickness; the analysis of the systolic function demonstrated a significant increase in ejection fraction, stroke volume, left ventricular mass, and end-diastolic volume in the athletes' population. Fluximetric study of transmitral and pulmonary venous flow showed that ventricular diastolic function is not influenced by hypertrophy. Our data indicate that diastolic function remains normal or improves in some cases after physical training; left ventricular hypertrophy and concentric remodeling do not involve diastolic changes like hypertrophic and hypertensive heart diseases. PMID- 11863322 TI - Separate and shared sources of dual-task cost in stimulus identification and response selection. AB - There is often strong interference if a second target stimulus (T2) is presented before processing of a prior target stimulus (T1) is complete. In the "Psychological Refractory Period" (PRP) paradigm, responses are speeded and interference manifests as increased response time for T2. In the "Attentional Blink" (AB) paradigm, stimuli are masked and responses unspeeded; interference manifests as reduced T2 accuracy. While different causes have usually been considered for PRP and AB phenomena, recent evidence has supported a unified account based on a single, shared restriction on concurrent processing. Here we show that a full assessment of separate and shared resource limitations requires direct comparison of hybrid PRP/AB trials with corresponding pure PRP and AB cases. Randomizing trial types in such a comparison also brings substantial benefit in addressing possible changes in task preparation or readiness. The data from two such experiments--combining speeded auditory (SA) and unspeeded visual (UV) task events--provide clear evidence for both separate and shared resource limitations. Often interference is strongest for T1 and T2 events of the same type, reflecting predominantly different limitations in SA and UV processing. With modest increases in demand, however, interference between different event types can also be made arbitrarily large, reflecting arbitrarily important shared limitations. For even such simple tasks as these, T--T2 interference reflects a combination of relatively local and relatively global sources. PMID- 11863323 TI - The interpolation of object and surface structure. AB - One of the main theoretical challenges of vision science is to explain how the visual system interpolates missing structure. Two forms of visual completion have been distinguished on the basis of the phenomenological states that they induce. Modal completion refers to the formation of visible surfaces and/or contours in image regions where these properties are not specified locally. Amodal completion refers to the perceived unity of objects that are partially obscured by occluding surfaces. Although these two forms of completion elicit very different phenomenological states, it has been argued that a common mechanism underlies modal and amodal boundary and surface interpolation (the "identity hypothesis"; Kellman & Shipley, 1991; Kellman, 2001). Here, we provide new data, demonstrations, and theoretical principles that challenge this view. We show that modal boundary and surface completion processes exhibit a strong dependence on the prevailing luminance relationships of a scene, whereas amodal completion processes do not. We also demonstrate that the shape of interpolated contours can change when a figure undergoes a transition from a modal to an amodal appearance, in direct contrast to the identity hypothesis. We argue that these and previous results demonstrate that modal and amodal completion do not result from a common interpolation mechanism. PMID- 11863324 TI - Web manuscript submission and tracking system for peer review. The journal moves with the time. PMID- 11863326 TI - Management of intermittent claudication: the importance of secondary prevention. AB - Atherosclerotic peripheral arterial disease (PAD) is a common disorder usually associated with silent or symptomatic arterial disease elsewhere in the circulation and a cluster of cardiovascular risk factors inducing atheroma progression and/or thrombotic complications. Because of these strong clinical associations, especially with coronary heart disease, the ankle-brachial pressure index (ABPI) is of prognostic significance. The clinical management of IC should include relief of symptoms combined with prevention of secondary cardiovascular complications, e.g. acute thrombotic events causing limb- or life-threatening ischaemia, which are often due to atherosclerotic plaque rupture leading to thrombotic vessel occlusion. Many patients with PAD do not receive an optimum package of secondary prevention, tailored to include maximum cholesterol reduction, BP and glycaemic control, ACE inhibition and single or combination anti-platelet therapy. This review considers recent information from large secondary prevention trials, e.g. the PAD subgroups within the HOPE, CAPRIE and statin studies. Slowing progression of atherosclerosis, and inducing stabilisation and regression of atheromatous plaques, is now feasible using long term combination drug therapy. The phrase angle quotation mark, leftangle quotation mark, leftconservative therapyangle quotation mark, rightangle quotation mark, right, popular among vascular surgeons, implies a passive minimal intervention strategy of surveillance and lifestyle advice; such terminology is perhaps no longer appropriate since considerable improvements in survival are likely to accrue if all patients with PAD, especially those with low ABPI, receive vigorous, titrated medical therapies, tailored to individual patients, as part of an evidence-based secondary prevention regime. PMID- 11863327 TI - Clinical and radiographic risk factors for operative stroke and death in the European carotid surgery trial. AB - OBJECTIVES: carotid endarterectomy is associated with significant morbidity and mortality. A better understanding of the relationships between baseline characteristics and outcome may help to reduce the risks of surgery. In order to make accurate and unbiased estimates of surgical risk it is important to study cohorts of patients that were established prospectively, where independent physicians assessed outcome, and where the decision to analyse and report the results was not data-dependent. The surgical arm of the European Carotid Surgery Trial (ECST) is such a cohort. METHODS: the 30-day outcome of carotid endarterectomy was analysed in ECST surgery patients in relation to their baseline clinical and angiographic characteristics. The severity of operative strokes was compared with that of strokes that occurred in the medical group. RESULTS: 1729 patients underwent trial surgery. There were 17 deaths (1.0%, 95% CI=0.6-1.6) and 105 non-fatal major strokes (6.1%, 95% CI=5.0-7.3) within 30 days of surgery. The risk of major stroke or death was 7.1% (95% CI=5.9-8.4). The risk of disabling or fatal stroke was 3.0% (95% CI=2.1-3.8). The ratio of disabling to non-disabling operative strokes was similar to that in the medical group. Several baseline characteristics predicted the operative risk of stroke and death in univariate analyses, but only four were independent risk factors in a multiple regression analysis: presentation with cerebral TIA vs ocular ischaemic events only (HR=2.99, 95% CI=1.33-6.69, p=0.008); female sex (HR=2.04, 95% CI=1.37- 3.06, p=0.001); systolic hypertension (HR=1.01 per 10 mmHg, 95% CI=1.00-1.02, p=0.03) and peripheral vascular disease (HR=2.17, 95% CI=1.17-2.89, p=0.001). CONCLUSIONS: the operative risk of stroke and death in the ECST was comparable with other prospective studies and trials in which patients were assessed postoperatively by both a physician and a surgeon. Case fatality and disability after operative stroke are similar to strokes that occur on medical treatment only. Several baseline patient characteristics predict surgical risk and it may be possible to use these characteristics to aid patient selection and surgical audit. PMID- 11863328 TI - Variation in surgical and anaesthetic technique and associations with operative risk in the European carotid surgery trial: implications for trials of ancillary techniques. AB - OBJECTIVES: several ancillary surgical techniques, such as shunting and patching, are used in association with carotid endarterectomy. However, the balance of risks and benefits of these techniques is uncertain because of the lack of large randomised controlled trials (RCTs). To assess the potential for further trials, we studied the variation in use of these techniques by surgeon and by country in the European Carotid Surgery Trial (ECST). METHODS: use of each ancillary technique was assessed by surgeon and by country. For each technique, the relationships between the use of the technique and baseline patient characteristics, use of other techniques, and the 30-day operative risk of stroke and death were determined. RESULTS: there was considerable variation between surgeons in the use of ancillary operative techniques both within (p<0.001 for shunting and patching), and between countries (p<0.001 for shunting and patching). Some surgeons used techniques selectively, and so the characteristics of patients differed depending on which techniques were used. Use of each technique was also significantly associated with the use of other techniques. Multiple regression analysis, taking into account all these factors, found no statistically significant associations between operative risk and the use of shunting, patching, intra-operative EEG monitoring, or type of anaesthetic. The only surgical technique significantly associated with an increased operative risk was not using intra-operative anticoagulation (hazard ratio=2.33, 95% CI=1.4 4.2). Other factors associated with an increased risk were an operation time of less than 1 h, or greater than 1.5 h, and the surgeons' subjective assessment that the operation was difficult. CONCLUSIONS: in the ECST, operative risk was more closely related to patient characteristics, length of surgery, and the surgeons' perception of the difficulty of the operation, than to the use of particular ancillary operative techniques. The considerable variation between surgeons, and between countries, in the use of ancillary techniques is in keeping with the lack of convincing data from RCTs, and suggests that there should be sufficient uncertainty to make large pragmatic trials possible. PMID- 11863329 TI - Systemic and local antibiotic prophylaxis in the prevention of prosthetic vascular graft infection: an experimental study. AB - AIM: to determine if local, in addition to systemic antibiotic prophylaxis (compared to that provided by systemic prophylaxis alone) provides additional benefit in terms of reducing graft infection. METHODS: gelatin-sealed Dacron grafts were interposed in the infrarenal aorta of 36 mongrels and inoculated with 1 ml of a S. aureus suspension. Group 1 (control group) received no prophylaxis and were inoculated with 1 ml containing 10(9)cfu/ml. Group 2 (n=6) received systemic prophylaxis (1 g cephamandole) and were inoculated with 10(5) cfu/ml (n=3) or 10(7) cfu/ml (n=3). Group 3 received systemic prophylaxis (1 g cephamandole) and were inoculated with 109 cfu/ml. Group 4 received systemic prophylaxis (2 g cephamandole) and were inoculated with 10(9)cfu/ml. In group 5 and 6 grafts were soaked in a rifampicin solution before use and inoculated with 10(9) cfu/ml. Group 5 received no systemic prophylaxis and group 6 received systemic prophylaxis (1 g cephamandole). Grafts were harvested at 2 weeks, and peritonitis, perigraft abscess, anastomotic disruption and graft occlusion recorded. Swabs were taken of the graft, the perigraft tissues and the peritoneal fluid. Graft segments were incubated in broth medium. RESULTS: inoculation with 10(9) cfu/ml ensured graft infection. Systemic or local prophylaxis alone failed to prevent graft infection. Only systemic and local antibiotic prophylaxis provided significant better results than no prophylaxis at all (p<0.01) and local prophylaxis alone (p<0.05). However, total "graft sterility" was not achieved as bacteriologic analysis of the graft segments showed low bacterial counts (<10 bacteria/graft) in 5 of 6 grafts. CONCLUSION: local and systemic prophylaxis provided more protection as demonstrated by the significant decrease in the incidence of "overt" graft infection. Total "graft sterility" cannot be expected in the case of an overwhelming bacterial challenge. PMID- 11863330 TI - The influence of gender difference on the outcomes of infrainguinal bypass for critical limb ischaemia in Chinese patients. AB - PURPOSE: to investigate the influence of gender difference on the outcomes of infrainguinal bypass operations performed in Chinese patients with critical limb ischaemia. METHODS: we prospectively studied the results of 191 consecutive infrainguinal bypass operations (98 men, 93 women) for critical lower limb ischaemia in Chinese patients. RESULTS: the women were older than men (median 75 vs 70 years, p=0.001) and cigarette smoking was commoner in men (83% vs 37% p<0.001). The calibre of run-off arteries at the level of distal anastomosis was smaller in women (median 2.5 mm vs 2.0 mm, p=0.03). The 30-day mortality was 3% (five men vs one woman, p=0.09) and early graft failure occurred in 19 patients (12 women vs seven men, p=0.28). At 3 months limb loss occurred in 16 (10 women vs six men, p=0.35) patients. Survival (38% vs 60% at 4 years, p=0.12) was similar in men and women. However, women suffered from poorer primary (33% vs 49% at 3 years, p=0.03) secondary graft patency rates (35% vs 64% at 3 years, p=0.02) than men. Limb survival rate in two groups (75% vs 85% at 4 years, p=0.18) was comparable. CONCLUSION: following infrainguinal bypass for critical limb ischaemia, early results were similar in both gender groups. In the long-term, women patients suffered from significantly higher graft failure rate. However, their long-term survival and limb salvage rate remained comparable to those of men. PMID- 11863331 TI - Duplex scanning as the sole preoperative imaging method for infrainguinal arterial surgery. AB - OBJECTIVE: to evaluate preoperative duplex as the sole investigation prior to lower limb reconstruction. Design retrospective analysis. MATERIALS AND METHODS: between January 1995 and December 1999, 157 of 329 surgical interventions for chronic infrainguinal arterial or aneurysmal disease were performed without preoperative angiography. RESULTS: in patients undergoing femoral artery endarterectomy, the extent of the stenosis and the status of the distal deep femoral artery were correctly diagnosed with duplex scanning in all but one patient. Duplex scan findings in patients undergoing infrainguinal bypass procedures were in agreement with the findings obtained from on-table angiography in regard to the selection of optimal outflow anastomotic sites in 123 (98%). Duplex scanning correctly evaluated the status of runoff in 113 (90%). There were no significant differences in 30-day occlusion rate and patency at 12 months between reconstructions performed with and without preoperative angiography. CONCLUSION: in patients with conclusive duplex scan findings there is no need to perform angiography prior to lower limb reconstruction. PMID- 11863332 TI - Treatment of renal artery fibromuscular dysplasia with balloon angioplasty: a prospective follow-up study. AB - OBJECTIVES: to assess restenosis rates and blood pressure response after percutaneous transluminal renal angioplasty (PTRA) in patients treated for fibromuscular dysplastic renal artery stenosis. METHODS: a prospective 12-month follow-up study of 27 patients with 31 treated renal artery stenosis. Follow-up assessment included colour-coded duplex sonography (CCD) of renal arteries, monitoring of blood pressure, antihypertensive medication, and creatinine measurements before discharge and at 3, 6, and 12 months. Primary end point was defined as a haemodynamically significant restenosis >60% assessed by CCD. RESULTS: there was a cumulative 23% restenosis rate at 12 months. Arterial hypertension was cured or improved in 93% of patients immediately after the intervention and remained cured/improved in 74% of patients at 12 months of follow-up. Renal failure present in five patients before PTRA stabilised or improved in all patients. CONCLUSION: although restenosis rate after PTRA in fibromuscular dysplasia is as high as in non-ostial atherosclerotic lesions, there remains a considerable higher therapeutic effect. Profound pressure response and recurrent arterial hypertension with restenosis support the high probability of a renovascular origin of arterial hypertension in this young and otherwise healthy population compared to patients with atherosclerotic renal artery lesions. PMID- 11863333 TI - Proteolysis of the abdominal aortic aneurysm wall and the association with rupture. AB - PURPOSE: to investigate proteolysis of the abdominal aortic aneurysm (AAA) wall and the association with rupture. METHODS: levels of matrix metalloproteinases (MMP-2 and MMP-9) and tissue inhibitor of metalloproteinases (TIMP-1 and TIMP-2) were measured in the walls of medium-sized (5-7 cm in diameter) ruptured AAA (rAAA) (n =30) and large (> or = 7 cm in diameter) asymptomatic AAA (aAAA) (n=30). RESULTS: MMP-2 levels (median, range) were significantly higher in the walls of large aAAA (165 ng/g AAA tissue, 50-840) than from medium-sized rAAA (110 ng/g AAA tissue, 47-547, p=0.007). MMP-9 levels were significantly higher in the walls of medium-sized rAAA (107 ng/g AAA tissue, 19-582) than from large aAAA (55 ng/g AAA tissue, 11-278, p=0.012). TIMP-1 and TIMP-2 levels were equivalent. There was a positive correlation between MMP-2 and the diameter of aAAA (r=0.54, p=0.002), but a negative correlation with MMP-9 (r= -0.44, p=0.017). No significant correlations were found between aAAA diameter and TIMP-1 or TIMP-2. CONCLUSION: AAA rupture is associated with higher levels of MMP-9. There is no association with TIMP-1 or TIMP-2 levels. MMP-2 levels are positively, whereas MMP-9 levels are negatively, correlated with aAAA size. MMP-9 may play a role in the progression towards rupture, whereas MMP-2 may play a role in expansion. PMID- 11863334 TI - Aortic aneurysm pulsatile wall motion imaged by cine MRI: a tool to evaluate efficacy of endovascular aneurysm repair? AB - OBJECTIVES: to evaluate cine MRI as a means of determining the two-dimensional pulsatile wall motion (2D-PWM) of abdominal aortic aneurysm (AAA). DESIGN: prospective study of 21 patients with AAA. 2D-PWM was defined as change in cross sectional area. RESULTS: the median diastolic area was 28 cm(2) (intraquartile range, IQR, 22-31 cm(2)) and the median (IQR) 2D-PWM was 0.25 (0.10-0.40) cm(2). Assuming that the AAA is circular in cross-section this represents a median (IQR) diameter increase of 0.3 (0.1-0.4) mm. However, local wall displacements up to 2 mm were present in varying directions, without significant change in surface area. CONCLUSION: AAA PWM is negligible and may not therefore be a potential tool to assess efficacy of endovascular aneurysm exclusion. PMID- 11863335 TI - The endovascular management of thoracic aortic disease -- some controversial issues. PMID- 11863336 TI - Endothelin receptors in the aetiology and pathophysiology of varicose veins. AB - INTRODUCTION: varicose veins are tortuous and poorly contractile. Their aetiology remains unclear. Neovascularisation has been suggested as a possible explanation. Endothelins are mitogenic, promoting proliferation and migration of endothelial cells via endothelin-B receptors. We hypothesise that endothelial cells and endothelin receptor density and distribution may play a role in the development of varicosis. METHODS: saphenous vein segments from nine patients with varicose veins were compared to six controls. Slide-mounted sections were incubated in radioactive labelled endothelin-1 and receptor subtype-selective ligands and binding sites assessed using autoradiography. Endothelin-1 and endothelial cells were identified by immunohistochemistry and CD31-positive staining cells counted. RESULTS: radioactive labelled endothelin-1 and endothelin-B receptor binding was reduced in varicose compared to control veins (p=0.04). Endothelin-A receptor binding was diffuse, with no difference in density in both groups (p=0.58). Endothelin-B receptor binding was diffuse with superimposed clusters. Although the density of medial endothelin-B receptor binding was reduced in the varicose group, more clusters were identified in this group compared to controls (p=0.005). CD-31 staining identified these clusters as endothelial cells. CONCLUSION: the reduced endothelin-1 binding and endothelin-B receptor density may be partially responsible for the reduced vasocontractility in varicose veins. We speculate that the increase in endothelin-B receptor binding CD31-positive endothelial cells in varicose veins may potentially stimulate mitogenesis and migration, leading to new vessel formation. PMID- 11863337 TI - Is there a place for external mesh wrapping of abdominal aortic aneurysms in the modern endovascular era? PMID- 11863338 TI - Sartorius transposition in the management of synthetic graft infection. PMID- 11863339 TI - Magnetic resonance angiography for postnephrectomy arteriovenous fistula. PMID- 11863340 TI - The technique of transilluminated powered phlebectomy -- a novel, minimally invasive system for varicose vein surgery. PMID- 11863342 TI - Pulmonary diffusion capacity as prognostic marker in chronic heart failure. PMID- 11863343 TI - Atrial fibrillation: one more sporting inconvenience? PMID- 11863344 TI - Tilt-induced asystole: a useful prognostic marker or clinically unrelevant finding? PMID- 11863345 TI - Upgrading from ventricular to physiological pacing: is it worth it? PMID- 11863346 TI - Clinical course after radiofrequency ablation of type I atrial flutter. Identification of patients who risk atrial arrhythmia recurrences. PMID- 11863347 TI - Oral glycoprotein IIb/IIIa receptor inhibitors in patients with cardiovascular disease: why were the results so unfavourable. PMID- 11863348 TI - A cardiovascular life history. A life course analysis of the original Framingham Heart Study cohort. AB - AIMS The objective of this paper is to measure the potential burden of cardiovascular disease within the original Framingham Heart Study cohort by transforming its well-described epidemiological measures into time-based health policy measures, such as life years lost to or lived with the disease. METHODS AND RESULTS We constructed multi-state life tables of the Framingham Heart Study cohort to calculate dwelling times with a history of cardiovascular disease. Age specific probabilities determined transitions from healthy through disease to death. For this synthetic cohort, from age 50 men (women) live on average 26 (32) years; 20 (26) free of cardiovascular disease. Allowing occupancy of more than one disease state, 50-year-old males (females) live 2 X 9 (1 X 2) years with a history of myocardial infarction, 0 X 93 (1 X 2) with a history of stroke, and 0 X 67 (0 X 93) with congestive heart failure. Having ever suffered acute myocardial infarction, stroke or congestive heart failure, life expectancy is reduced by 9 (13), 12 (15) or 16 (16) years, respectively in 60-year-old men (women). CONCLUSIONS Transforming occurrence probabilities into time-based health measures, the prevalence of cardiovascular disease is remarkable: from age 50, 20% of remaining life expectancy is lived with the disease. Such measures are integral to appropriate health planning and assessment of the potential population health value of various treatment and prevention strategies. PMID- 11863349 TI - Alveolar--capillary membrane gas conductance: a novel prognostic indicator in chronic heart failure. AB - AIMS: Lung dysfunction occurring in chronic heart failure worsens clinical status and exercise performance. The prognostic value of airway and alveolar function measurements in chronic heart failure has not been explored. We aimed to evaluate the prognostic value of lung function tests in a population of patients with stable chronic heart failure. METHODS AND RESULTS: One hundred and six stable chronic heart failure patients (whose left ventricular ejection fraction averaged 33 +/- 1%) underwent echocardiography, metabolic stress testing, assessment of pulmonary function at rest (by spirometry), of alveolar diffusing capacity (DLco) (with carbon monoxide technique) and of its membrane (DM) and capillary blood volume (Vc) components. Prognostic relevance of pulmonary variables was assessed by the Kaplan-Meier approach with log-rank testing and by Cox regression analysis. Cut-off values of lung parameters were based on the 33rd and 66th centiles. Seventeen patients died for cardiac reasons. Non-survivors compared to survivors showed lower forced expiratory volume in 1 s (2 X 1 +/- 0 X 1 vs 2 X 4 +/- 0 X 1 l; P<0 X 01), forced vital capacity (2 X 6 +/- 0 X 1 vs 2 X 9 +/- 0 X 1 l; P<0 X 01), maximal voluntary ventilation (80 X 7 +/- 2 X 5 vs 95 X 4 +/- 2 X 7 l; P<0 X 01), DLco (16 X 5 +/- 1 X 1 vs 19 X 3 +/- 0 X 6 ml . min(-1) . mmHg(-1); P<0 X 01) and DM (25 X 1 +/- 1 X 8 vs 31 X 9 +/- 1 X 5 ml . min(-1) . mmHg(-1); P<0 X 01). They also exhibited a smaller peak VO2 (14 X 6 +/- 0 X 7 vs 15 X 9 +/- 0 X 6 ml . min(-1) . kg(-1); P<0 X 05) and a steeper VE/VCO2 slope (45 X 0 +/- 1 X 7 vs 41 X 9 +/- 1 X 5; P<0 X 01). Multivariate analysis revealed that DM was the only independent predictor of cardiac death. Cases at high risk for adverse outcome were identified by a DM<24 X 7 ml . min(-1) . mmHg(-1). Patients receiving ACE-inhibitors presented with a higher DM (32 X 1 +/- 1 X 7 vs 27 X 9 +/- 1 X 7 ml . min(-1) . mmHg(-1), P<0 X 05) as well as a better Cox estimated survival rate. CONCLUSIONS: Impaired DM is a powerful independent predictor of worse prognosis in stable chronic heart failure and may be considered an additional index of disease severity, as well as a specific therapeutic target. PMID- 11863350 TI - Long-lasting sport practice and lone atrial fibrillation. AB - AIMS: To analyse whether the proportion of patients with lone atrial fibrillation engaged in chronic sport practice was higher than that observed in the general population. METHODS AND RESULTS: The records of 1160 patients, seen at the arrhythmia outpatient clinic, were reviewed. A total of 70 patients (6%) suffered lone atrial fibrillation and were younger than 65 years. Thirty two of them had been engaged in long-term sport practice. All patients in the sport group were men as compared to only 50% in the sedentary group (P<0 x 0001). To avoid the confounding effect of sex distribution, women were excluded. Sportsmen started their episodes of atrial fibrillation at a younger age, they had a lower incidence of mild hypertension and their episodes of atrial fibrillation were predominantly vagal in contrast to the sedentary patients. The echocardiographic parameters were similar to those observed in the sedentary patients, but when compared with 20 healthy controls, they showed greater atrial and ventricular dimensions and a higher ventricular mass. The proportion of sportsmen among patients with lone atrial fibrillation is much higher than that reported in the general population of Catalonia: 63% vs 15% (P<0 x 05). CONCLUSION: Long-term vigorous exercise may predispose to atrial fibrillation. PMID- 11863351 TI - Long-term outcome of patients with asystole induced by head-up tilt test. AB - AIMS: To analyse the long-term outcome of the largest reported cohort of patients presenting asystole during head-up tilt test. METHODS AND RESULTS: Since 1990, 1322 patients with syncope of unknown origin have undergone tilt-table testing. Of those, 330 patients (24 X 9%) presented an abnormal response (syncope or pre syncope). Furthermore, 58 of those patients (17 X 5%) suffered a period of asystole (> or = 3000 ms) during the test. Asystole (median (interquartile range)) lasted 10 (4, 19 X 2) s (range 3-90). Two different protocols (angles) of tilting (Westminster (60 degrees) n=1124; isoproterenol (80 degrees) n=198)) influenced the time to the syncopal episode (13 (6 X 5, 20 X 5) vs 2 (1, 6 X 5) min, P=0,0005) but not the duration of the asystole. During this period, therapy for asystole featured three different stages: first patients were treated with pacemakers; later drug therapy (metoprolol and/or etilefrine) was recommended; lastly (from 1995), no specific treatment was given. In a cohort age- and gender matched study, those patients without were compared to those with asystole in a 2:1 basis. During 40 X 7 months of follow-up (17 X 7, 66 X 8), 12 patients (20 X 6%) with asystole had syncopal recurrences. Furthermore, 34 patients (28 X 8%) without asystole presented syncopal episodes during a follow-up of 51 X 6 months (29 X 3, 73 X 1) (P=ns). The Kaplan-Meier analysis in patients with and without asystole showed a mean time free of recurrence of 92 X 6 +/- 6 months vs 82 X 6 +/- 4 X 7 months (P=ns). The previous number of syncopes had a significant relationship with recurrences (P=0 X 002), but not therapy. There were no cardiac related deaths. CONCLUSIONS: (1) Asystole during head-up tilt test does not imply a malignant outcome and syncope recurrence is low; (2) pacemaker or drug therapy do not significantly influence outcome which correlates to the previous number of syncopal episodes but not to gender, age, asystole occurrence, asystole duration and timing to asystole during head-up tilt test; (3) tilting protocol (angle) might influence time to and incidence of asystole during head-up tilt test. PMID- 11863352 TI - Improved cardiac function and quality of life following upgrade to dual chamber pacing after long-term ventricular stimulation. AB - AIMS: Many patients with sinus node disease or atrioventricular block have previously received pacemakers with only ventricular stimulation (VVI or VVIR). This study aimed to investigate whether quality of life and cardiac function were affected by an upgrade to dual chamber pacing (DDDR or DDIR) following long-term ventricular stimulation. METHODS: After implantation of an atrial lead and a DDDR pulse generator, a randomized, double-blind crossover study was performed in 19 patients, previously treated with ventricular pacing for a median time of 6 X 8 years. Patients were randomized to 8 weeks with either VVIR or DDDR/DDIR pacing; after this time, the other mode was programmed for 8 weeks. At the end of each period, the patients' quality of life was evaluated and echocardiography was performed together with Holter monitoring and blood samples for brain natriuretic peptide. RESULTS: Sixteen of the patients preferred DDDR and two VVIR pacing (P=0 X 001); one was undecided. Seven patients demanded an early crossover while paced in the VVIR mode, vs none in the DDDR mode (P=0 X 008). Quality of life was higher in the DDDR mode in 11 of 17 modalities, reaching statistical significance for dyspnoea (P<0 X 05) and general activity (P<0 X 05). Echocardiography showed significantly larger left ventricular end-diastolic dimensions in the DDDR mode (P=0 X 01), whereas end-systolic dimensions did not differ. Left ventricular systolic function was significantly superior in the DDDR mode (mean aortic velocity-time integral: P<0 X 001) and left atrial diameter was significantly smaller in the DDDR mode (P=0 X 01). The plasma level of brain natriuretic peptide was significantly lower in DDDR mode (P=0 X 002). CONCLUSION: An upgrade to dual chamber rate adaptive pacing results in significantly improved quality of life and cardiac function as compared to continued VVIR stimulation and should thus be considered in patients with ventricular pacemakers who have not developed permanent atrial fibrillation or flutter. PMID- 11863353 TI - Factors associated with early atrial fibrillation after ablation of common atrial flutter. A single centre prospective study. AB - BACKGROUND: The occurrence of early atrial fibrillation (< or = 6 months) after ablation of common atrial flutter is of clinical significance. Variables predicting this evolution in ablated patients without a previous atrial fibrillation history have not been fully investigated. OBJECTIVES: The aim of the present study was: (1) to identify predictive factors of early atrial fibrillation (< or = 6 months) in the overall population following atrial flutter catheter ablation; (2) to identify predictive variables of early atrial fibrillation following (< or = 6 months) atrial flutter catheter ablation within a subgroup of patients without documented prior atrial fibrillation. METHODS: This study prospectively included 96 consecutive patients (age 65 +/- 13 years; 18 women) over a 12-month period. Their counterclockwise flutter was ablated by radiofrequency, by the same operator, with an 8-mm-tip catheter. Clinical, electrophysiological and echocardiographic data were collected and 27 variables were retained for analysis: age; gender; type of atrial flutter (permanent vs paroxysmal); symptom duration (months +/- SD); pre-ablation history of atrial fibrillation; structural heart disease; left ventricular ejection fraction (%); left atrial size (mm); cava--tricuspid isthmus dimension; septal isthmus dimension; systolic pulmonary pressure > or < or = 30 mmHg; right atrial area; left atrial area; isthmus block; number of radiofrequency applications (+/- SD); antiarrhythmic drugs at discharge; left ventricular diastolic diameter; left ventricular systolic diameter; left ventricular telediastolic volume; left ventricular telesystolic volume; A-wave velocity (cm . s(-1)); E-wave velocity (cm . s(-1)); E/A; isovolumetric relaxation time; E-wave deceleration time; significant mitral regurgitation and flutter cycle length (ms). RESULTS: Of the 96 consecutive ablated patients, early atrial fibrillation was documented in 16 patients (17%). Atrial fibrillation occurred 30 +/- 46 days (range 1 to 171 days) after ablation. Univariate analysis associated an early occurrence of atrial fibrillation with: atrial fibrillation history, left ventricular ejection fraction, left atrial size, left ventricular telesystolic volume, A-wave velocity, significant mitral regurgitation and flutter cycle length. Multivariate analysis using a Cox model found that the only independent predictors of early atrial fibrillation were left ventricular ejection fraction and pre-ablation history of atrial fibrillation. In the subgroup without prior atrial fibrillation history (n=63; 66%), the only independent predictor of early atrial fibrillation was the presence of a significant mitral regurgitation. CONCLUSIONS: In a subgroup of patients without atrial fibrillation history, 8% of patients revealed an early atrial fibrillation. Mitral regurgitation is a strong predictive factor of early atrial fibrillation occurrence with 80% sensitivity, 78% specificity and 98% negative predictive value. These data should be considered in post-ablation management. PMID- 11863355 TI - In silico biology: more than computer games. PMID- 11863356 TI - Physical and transcript map of a 2-Mb region in Xp22.1 containing candidate genes for X-linked mental retardation and short stature. AB - Genetic loci for several diseases, including X-linked nonspecific mental retardation and short stature, have been mapped to Xp22.1. In spite of the recent publications of two draft sequences for the human genome, this region seems to be largely unmapped and unsequenced. Here we report an integrated physical and transcript map of approximately 2-Mb from DXS8004 to DXS365. Using sequence tagged site (STS)-content mapping and chromosome walking, we assembled a genomic clone contig of 54 BACs and one cosmid with an estimated 4.5-fold coverage of this region. The minimum tiling path consists of 23 BACs and one cosmid. Onto this contig, we mapped 30 new STSs derived from the unique end-sequences of the BACs, three expressed sequence tags, five genes, and seven CpG islands. This integrated map provides a unique resource for the positional cloning of candidate disease genes mapping to Xp22.1 and is therefore of value for the completion of the genomic sequence of this region. PMID- 11863358 TI - An anthropoid-specific locus of orphan C to U RNA-editing enzymes on chromosome 22. AB - The cytidine (C) to uridine (U) editing of apolipoprotein (apo) B mRNA is mediated by tissue-specific, RNA-binding cytidine deaminase APOBEC1. APOBEC1 is structurally homologous to Escherichia coli cytidine deaminase (ECCDA), but has evolved specific features required for RNA substrate binding and editing. A signature sequence for APOBEC1 has been used to identify other members of this family. One of these genes, designated APOBEC2, is found on chromosome 6. Another gene corresponds to the activation-induced deaminase (AID) gene, which is located adjacent to APOBEC1 on chromosome 12. Seven additional genes, or pseudogenes (designated APOBEC3A to 3G), are arrayed in tandem on chromosome 22. Not present in rodents, this locus is apparently an anthropoid-specific expansion of the APOBEC family. The conclusion that these new genes encode orphan C to U RNA editing enzymes of the APOBEC family comes from similarity in amino acid sequence with APOBEC1, conserved intron/exon organization, tissue-specific expression, homodimerization, and zinc and RNA binding similar to APOBEC1. Tissue-specific expression of these genes in a variety of cell lines, along with other evidence, suggests a role for these enzymes in growth or cell cycle control. PMID- 11863359 TI - Efficiency of de novo centromere formation in human artificial chromosomes. AB - In a comparative study, we show that human artificial chromosome (HAC) vectors based on alpha-satellite (alphoid) DNA from chromosome 17 but not the Y chromosome regularly form HACs in HT1080 human cells. We constructed four structurally similar HAC vectors, two with chromosome 17 or Y alphoid DNA (17alpha, Yalpha) and two with 17alpha or Yalpha and the hypoxanthine guanine phosphoribosyltransferase locus (HPRT1). The 17alpha HAC vectors generated artificial minichromosomes in 32-79% of the HT1080 clones screened, compared with only approximately 4% for the Yalpha HAC vectors, indicating that Yalpha is inefficient at forming a de novo centromere. The 17alpha HAC vectors produced megabase-sized, circular HACs containing multiple copies of alphoid fragments (60 250 kb) interspersed with either vector or HPRT1 DNA. The 17alpha-HPRT1 HACs were less stable than those with 17alpha only, and these results may influence the design of new HAC gene transfer vectors. PMID- 11863360 TI - Identification of a psoriasis susceptibility candidate gene by linkage disequilibrium mapping with a localized single nucleotide polymorphism map. AB - Psoriasis is a chronic inflammatory disease of the skin with both genetic and environmental risk factors. Here we describe the creation of a single-nucleotide polymorphism (SNP) map spanning 900-1200 kb of chromosome 3q21, which had been previously recognized as containing a psoriasis susceptibility locus, PSORS5. We genotyped 644 individuals, from 195 Swedish psoriatic families, for 19 polymorphisms. Linkage disequilibrium (LD) between marker and disease was assessed using the transmission/disequilibrium test (TDT). In the TDT analysis, alleles of three of these SNPs showed significant association with disease (P<0.05). A 160-kb interval encompassing these three SNPs was sequenced, and a coding sequence consisting of 13 exons was identified. The predicted protein shares 30-40% homology with the family of cation/chloride cotransporters. A five marker haplotype spanning the 3' half of this gene is associated with psoriasis to a P value of 3.8<10(-5). We have called this gene SLC12A8, coding for a member of the solute carrier family 12 proteins. It belongs to a class of genes that were previously unrecognized as playing a role in psoriasis pathogenesis. PMID- 11863361 TI - Molecular cloning of ADIR, a novel interferon responsive gene encoding a protein related to the torsins. AB - The expression of the previously uncharacterized gene Adir (for ATP dependent interferon responsive gene) was increased by 5- to 15-fold in tissue of the oral cavity or in spleen and liver of mice treated orally or intraperitoneally with IFN-alpha, and in mouse cells treated in vitro with IFN-alpha or IFN-gamma. The level of Adir mRNA was also increased 20- to 40-fold in the brains of animals infected with encephalomyocarditis virus. Adir is expressed ubiquitously in mouse tissues as 1.9-, 2.4-, and 3.5-kb mRNA transcripts encoding a 385-amino-acid protein with a conserved ATP binding domain containing typical nucleotide and Mg(2+) binding sites. We also characterized the human ortholog, ADIR, which is located on chromosome 1q25-q31 and contains six exons encoding a 397-amino-acid protein with 80% homology to the mouse protein. A single 2.3-kb mRNA was detected in all human tissues examined, except for placenta, which also contained a 1.25 kb tissue-specific transcript generated by alternative splicing and encoding a putative 336-amino-acid protein. Although ADIR exhibits low homology to DYT1 and TOR1B, the deduced ADIR protein sequences are highly homologous to torsin A and torsin B and more distantly related to members of the Clp/HSP100 family of proteins, suggesting that ADIR, like torsins, is related to the AAA chaperone like family of ATPases. An ADIR-EGFP fusion protein expressed in HeLa cells was shown to be associated with the endoplasmic reticulum. PMID- 11863362 TI - A method for developing high-density SNP maps and its application at the type 1 angiotensin II receptor (AGTR1) locus. AB - Evaluating the potential genetic components of complex disease will likely be aided through the use of dense polymorphism maps. Previously, we reported evidence for linkage with diabetic nephropathy on chromosome 3q in a region encompassing the type 1 angiotensin II receptor (AGTR1) gene. To further investigate any role for this gene in disease onset, we set out to design a dense polymorphism map spanning the AGTR1 locus for the purpose of association studies. Toward this goal, we have developed a technique for rapid identification of polymorphisms in long stretches of genomic DNA. This approach uses long-range PCR, DNA pooling, and transposon-based DNA sequencing. Using this technique, we efficiently validated and genotyped 18 polymorphisms spanning the 60.5-kb AGTR1 locus. Our panel of polymorphisms has an average spacing of 3.2 kb and an average minor allele frequency of 24%. PMID- 11863364 TI - Genomic organization and mapping of the gene encoding the PP2A B56gamma regulatory subunit. AB - Protein phosphatase 2A (PP2A) is a major serine/threonine phosphatase that regulates a wide variety of cellular processes. The enzymatic activity and intracellular localization of PP2A are determined by three distinct families of cellular regulatory subunits (B, B'', and B''). The B' subunit, also known as B56, is the most diverse, consisting of five isoforms (alpha, beta, gamma, delta, and epsilon). The gene encoding B56gamma has been designated as PPP2R5C and encodes three differentially spliced variants: B56gamma1, -gamma2, and -gamma3. However, conflicting chromosomal loci have been reported in human genomic databases. The original cytogenetic mapping placed the gene on chromosome 3p21.3, whereas subsequent studies using radiation hybrid analysis localized PPP2R5C to chromosome 14q. In this study, by radiation hybrid mapping, FISH analysis, BAC clone sequencing, and RT-PCR analysis, we show that the functional gene PPP2R5C exists at 14q32.2 and gives rise to three splicing variants, B56gamma1, -gamma2, and -gamma3, whereas a nonfunctional B56gamma1 pseudogene, PPP2R5CP, is present at 3p21.3. We also report the genomic organization of both the functional gene and the pseudogene. PMID- 11863363 TI - Novel paralogy relations among human chromosomes support a link between the phylogeny of doublesex-related genes and the evolution of sex determination. AB - Recent advances in the evolutionary genetics of sex determination indicate that DMRT1 may be a vertebrate equivalent of the Drosophila melanogaster master sex regulator gene, doublesex. The role of DMRT1 seems to be confined to some aspects of male sex differentiation, whereas in Drosophila, doublesex has wider developmental effects in both sexes. This suggests other homologs of doublesex may exist in the vertebrate genome and encode sex-specific functions not displayed by DMRT1. We identified and characterized five novel human DM genes, distinct from previously described family members. Human DM genes map to three well-defined regions of chromosomes 1, 9, and 19 (one gene on chromosome 19 having an additional homolog on chromosome X). We collated data indicating these chromosomal regions harbor multiple syntenic genes sharing highly specific paralogy relations, suggesting that they arose early during vertebrate evolution. The 9p21-p24.3 bands represent the ancestral copy and harbor closely linked DM genes that may reflect the overall diversity of the fruit fly DM gene family. The human genome contains a small number of potential doublesex homologs that may be involved in human sexual development. Identifying highly conserved chromosomal regions, such as distal 9p, is an important tool to trace complex ancient evolutionary processes inaccessible by other approaches. PMID- 11863365 TI - Comparison of human chromosome 6p25 with mouse chromosome 13 reveals a greatly expanded ov-serpin gene repertoire in the mouse. AB - Ov-serpins are intracellular proteinase inhibitors implicated in the regulation of tumor progression, inflammation, and cell death. The 13 human ov-serpin genes are clustered at 6p25 (3 genes) and 18q21 (10 genes), and share common structures. We show here that a 1-Mb region on mouse chromosome 13 contains at least 15 ov-serpin genes compared with the three ov-serpin genes within 0.35 Mb at human 6p25 (SERPINB1 (MNEI), SERPINB6 (PI-6), SER-PINB9 (PI-9)). The mouse serpins have characteristics of functional inhibitors and fall into three groups on the basis of similarity to MNEI, PI-6, or PI-9. The genes map between the mouse orthologs of the Werner helicase interacting protein and NAD(P)H menadioine oxidoreductase 2 genes, in a region that contains the markers D13Mit136 and D13Mit116. They have the seven-exon structure typical of human 6p25 ov-serpin genes, with identical intron phasing. Most show restricted patterns of expression, with common sites of synthesis being the placenta and immune tissue. Compared with human, this larger mouse serpin repertoire probably reflects the need to regulate a larger proteinase repertoire arising from differing evolutionary pressures on the reproductive and immune systems. PMID- 11863366 TI - Methylation patterns of the human beta-glucuronidase gene locus: boundaries of methylation and general implications for frequent point mutations at CpG dinucleotides. AB - Methylation of CpG islands spanning promoter regions is associated with control of gene expression, although it is unclear what mechanisms define the boundaries between methylated and unmethylated regions in the genome. Methylation of genomic DNA in mammals also affects the frequency of inherited diseases by predisposing them to CpG mutations. To gain insight into these issues, we investigated patterns of cytosine methylation on almost the entire beta-glucuronidase gene (GUSB) from normal leukocyte DNAs by bisulfite genomic sequencing. We mapped the boundaries of methylation that flank the 5'- and 3'-ends of the CpG island region, and correlated methylation status with transitional mutations at CpG sites. GenBank sequence analyses showed that the CpG island of human GUSB is juxtaposed with multiple Alu repeats and also includes multiple Sp1 sites upstream and downstream of the transcription start, which has been suggested to prevent CpG islands from becoming methylated. We show that cytosine methylation is extensive across the entire gene except for CpG sites in the proximal promoter region, exon 1, and part of intron 1; the unmethylated CpG island is embedded between densely methylated flanking regions containing multiple Alu repeats; a sharp boundary separates the methylated and unmethylated regions of the 5'-flank of the CpG island, but a gradual change in methylation density over 1.0 kb is observed in the 3'-flank of the CpG island; boundaries of the 5'-end and 3'-end of the CpG island contain multiple Sp1 sites in addition to Alu repeats; methylation in both strands is symmetrical except at the boundary regions between methylated and unmethylated regions; and nonmethylation of exon 1 correlates with the absence of transitional mutations at CpG sites in exon 1. PMID- 11863367 TI - The sequence, expression, and chromosomal localization of a novel polycystic kidney disease 1-like gene, PKD1L1, in human. AB - Polycystin-1 and polycystin-2 are the products of PKD1 and PKD2, genes that are mutated in most cases of autosomal dominant polycystic kidney disease. Since the first two polycystins were cloned, three new members, polycystin-L, -2L2, and REJ, have been identified. In this study, we describe a sixth member of the family, polycystin-1L1, encoded by PKD1L1 in human. The full-length cDNA sequence of PKD1L1, determined from human testis cDNA, encodes a 2849-amino-acid protein and 58 exons in a 187-kb genomic region. The deduced amino acid sequence of polycystin-1L1 has significant homology with all known polycystins, but the longest stretches of homology were found with polycystin-1 and -REJ over the 1453 and 932-amino-acid residues, respectively. Polycystin-1L1 is predicted to have two Ig-like PKD, a REJ, a GPS, a LH2/PLAT, a coiled-coil, and 11 putative transmembrane domains. Several rhodopsin-like G-protein-coupled receptor (GPCR) signatures are also found in polycystin-1L1. Dot-blot analysis and RT-PCR revealed that human PKD1L1 is expressed in testis and in fetal and adult heart. In situ hybridization analysis showed that the most abundant and specific expression of Pkd1l1 was found in Leydig cells, a known source of testosterone production, in mouse testis. We have assigned PKD1L1 to the short arm of human chromosome 7 in bands p12--p13 and Pkd1l1 to mouse chromosome 11 in band A2 by fluorescence in situ hybridization. We hypothesize a role for polycystin-1L1 in the heart and in the male reproductive system. PMID- 11863357 TI - Identification of target genes regulated by PAX3 and PAX3-FKHR in embryogenesis and alveolar rhabdomyosarcoma. AB - PAX3 is a transcription factor important for neural, muscle, and facial development in vertebrates. To identify genes regulated by PAX3, we used a cyclic amplification and selection of targets (CASTing) strategy to isolate cis regulatory elements bound by PAX3. CASTing libraries were constructed with mouse DNA fragments bound by mouse PAX3, and human genomic DNA fragments bound by human PAX3 and the fusion protein PAX3-FKHR. Approximately 1000 clones were sequenced from each of these three libraries. Numerous putative targets of PAX3 and PAX3 FKHR were identified and six genes, Itm2A, Fath, FLT1, TGFA, BVES, and EN2, were examined closely. The genomic DNA fragments near these genes contain PAX3 binding sites and confer PAX3-dependent regulation. The expression levels of these genes correlate with the PAX3 expression levels in mouse embryos or with PAX3-FKHR expression levels in rhabdomyosarcoma cell lines, and indicate they may be part of the PAX3 regulatory circuitry during embryogenesis and tumor formation. PMID- 11863368 TI - Discovery of a functional retrotransposon of the murine phospholipid hydroperoxide glutathione peroxidase: chromosomal localization and tissue specific expression pattern. AB - Phospholipid hydroperoxide glutathione peroxidase (PHGPx), a selenoprotein capable of reducing toxic hydroperoxy ester lipids, has been implicated in antioxidative defense and spermatogenesis. Screening a murine genomic library, we isolated two recombinants (pseudogenes 1 and 2) containing retrotransposons for this enzyme. On comparison with the paralogous cDNA, pseudogene 1 contained only two silent nucleotide exchanges, and the 3'-untranslated region (3'-UTR) carrying the functionally important selenocysteine insertion sequence was free of mutations. This retrotransposon was found in various mouse strains and could be mapped to the region B2-B3 of chromosome 10. In vitro studies indicated significant promoter activity in the 5'-flanking region of pseudogene 1, and we observed a tissuespecific expression of this retrotransposon. In the submandibular gland. Most PHGPx transcripts originated from pseudogene 1. In contrast, pseudogene 2, containing numerous mutations in all parts of the retrotransposon, was not expressed in any tissue. It was mapped to region E3-E4 of chromosome 17, and we did not detect any promoter activity in its 5'-flanking region. These data indicate the existence of two retrotransposed PHGPx pseudogenes, one of which encodes a functional enzyme. This retrotransposon belongs to the rare group of pseudogenes that are tissue-specifically expressed under the control of captured regulatory elements, and it constitutes an example of evolutionarily acquired redundancy in gene expression. The results are important for the design of future knockout strategies aimed at investigating the biological role of this enzyme. PMID- 11863369 TI - Genomic organization and characterization of the human type XXI collagen (COL21A1) gene. AB - We cloned a 4.1-kb full-length cDNA based on a reported human genomic clone containing a partial open reading frame (ORF) coding for a novel collagen-like protein. Sequence analysis indicated that the ORF codes for the alpha(1)-chain of type XXI collagen. Assembly of the genomic data reveals a complete sequence of the human gene COL21A1. COL21A1 is localized to chromosome 6p11.2-12.3, spanning 337 kb in size. The gene contains 31 exons, in which the 5'-untranslated exons 1 and 1a are alternatively spliced. The exon/domain organization of COL21A1 resembles that of the reported FACIT collagen genes, including COL9A1, COL9A2, COL9A3, and COL19A1, suggesting that these genes may have derived from the same ancestor FACIT gene by duplication. The expression of COL21A1 in human tissues is developmentally regulated, with a higher level at fetal stages. Type XXI collagen is an extracellular matrix component of the blood vessel walls, secreted by smooth-muscle cells. Platelet-derived growth factor (PDGF) has a pronounced effect on the stimulation of COL21A1 expression in cultured aortic smooth-muscle cells, suggesting that alpha1(XXI) collagen may contribute to the extracellular matrix assembly of the vascular network during blood vessel formation. PMID- 11863370 TI - BHLHB4 is a bHLH transcriptional regulator in pancreas and brain that marks the dimesencephalic boundary. AB - We have cloned a basic helix-loop-helix (bHLH) factor gene, Bhlhb4, from a mouse beta-cell line. Fluorescence in situ hybridization (FISH) and genetic mapping place Bhlhb4 at the telomeric end of mouse chromosome 2 (H3-H4), syntenic to human chromosome 20q13. Based on phylogenetic analysis, BHLHB4 belongs to a new subgroup of bHLH factors including at least four previously identified mouse bHLH factors: BHLHB5, MIST1, OLIG1, OLIG2, and OLIG3. In the developing nervous system, Bhlhb4 was found to mark the dimesencephalic boundary, suggesting that Bhlhb4 may have a role in diencephalic regionalization. In the pancreas, Bhlhb4 is expressed in a transient fashion that suggests a role in the pancreatic endocrine cell lineage. Transfection experiments show that BHLHB4 can repress transcriptional activation mediated through the pancreatic beta-cell specific insulin promoter enhancer RIPE3. Together, these data suggest that BHLHB4 may modulate the expression of genes required for the differentiation and/or maintenance of pancreatic and neuronal cell types. PMID- 11863371 TI - A technique for genome-wide identification of differences in the interspersed repeats integrations between closely related genomes and its application to detection of human-specific integrations of HERV-K LTRs. AB - We have developed a method of targeted genomic difference analysis (TGDA) for genomewide detection of interspersed repeat integration site differences between closely related genomes. The method includes a whole-genome amplification of the flanks adjacent to target interspersed repetitive elements in both genomic DNAs under comparison, and subtractive hybridization (SH) of the selected amplicons. The potential of TGDA was demonstrated by the detection of differences in the integration sites of human endogenous retroviruses K (HERV-K) and related solitary long terminal repeats (LTRs) between the human and chimpanzee genomes. Of 55 randomly sequenced clones from a library enriched with human-specific integration (HSI) sites, 33 (60%) represented HSIs. All the human-specific (Hs) LTRs belong to two related evolutionarily young groups, suggesting simultaneous activity of two master genes in the hominid lineage. No deletion/insertion polymorphism was detected for the LTR HSIs for 25 unrelated caucasoid individuals. We also discuss the possible research applications for TGDA research. PMID- 11863372 TI - NAPP2, a peroxisomal membrane protein, is also a transcriptional corepressor. AB - Nuclear factor-erythroid number 2 (NF-E2) is a positive regulatory, DNA binding transcription factor for gene expression in erythroid and megakaryocytic cells. To further understand the mechanisms of NF-E2 function, we used expression cloning to identify coregulators interacting with the erythroid-specific subunit of NF-E2, p45. We have isolated a protein, NAPP2, which contains an aspartic-acid and glutamic-acid-rich region and a nuclear localization signal. The gene encoding NAPP2, PEX14, is located on chromosome 1p36 and is ubiquitously expressed. The domains of interaction in vitro and in vivo between p45 and NAPP2 were mapped by a yeast two-hybrid system and cotransfection experiments. In mammalian cell culture, ectopically expressed NAPP2 inhibited p45-directed transcriptional activation. Furthermore, NAPP2 functions as a corepressor and interacts specifically with histone deacetylase l (HDAC1), but not HDAC2 or HDAC3. NAPP2 is thus potentially a negative coregulator of NF-E2. NAPP2 is identical to PEX14, an integral membrane protein essential for protein docking onto the peroxisomes. These studies have identified a novel, bifunctional protein capable of acting as a transcriptional corepressor and a polypeptide transport modulator. They also suggest that NF-E2 may function both positively and negatively in the transcription regulation of specific erythroid and megakaryocytic genes. PMID- 11863373 TI - A BAC contig map of the Ly49 gene cluster in 129 mice reveals extensive differences in gene content relative to C57BL/6 mice. AB - The murine Ly49 gene family is functionally analogous to the human killer cell Ig like receptor (KIR) family of class I major histocompatibility complex (MHC) receptors. The number of KIR genes varies dramatically between individuals; however, the organization of the Ly49 genes has only been determined for the C57BL/6 (B6) mouse. The organization of the 129 Ly49 loci was determined from a BAC contig map by PCR and Southern blot analysis. In addition to the 10 Ly49 genes known from previous studies of the 129/J strain, 8 new genes were localized to the 129 Ly49 cluster. A gene order of Ly49q(1), e, (v, q(2)), e/c(2), l/r, s, t, e/c(1), r, u, u/i, i(1), g, p/d, (i(2), p), and o was determined. The 129 Ly49 gene cluster is predicted to span approximately 600 kb. These results indicate that Ly49 gene numbers can be significantly different between inbred mouse strains, analogous to the haplotype differences observed in the human KIR genes. PMID- 11863376 TI - Conservation of gene order between horse and human X chromosomes as evidenced through radiation hybrid mapping. AB - A radiation hybrid (RH) map of the equine X chromosome (ECAX) was obtained using the recently produced 5000(rad) horse x hamster hybrid panel. The map comprises 34 markers (16 genes and 18 microsatellites) and spans a total of 676 cR(5000), covering almost the entire length of ECAX. Cytogenetic alignment of the RH map was improved by fluorescent in situ hybridization mapping of six of the markers. The map integrates and refines the currently available genetic linkage, syntenic, and cytogenetic maps, and adds new loci. Comparison of the physical location of the 16 genes mapped in this study with the human genome reveals similarity in the order of the genes along the entire length of the two X chromosomes. This degree of gene order conservation across evolutionarily distantly related species has up to now been reported only between human and cat. The ECAX RH map provides a framework for the generation of a high-density map for this chromosome. The map will serve as an important tool for positional cloning of X-linked diseases/conditions in the horse. PMID- 11863375 TI - cDNA cloning, genomic structure, chromosomal mapping, and functional expression of a novel human alanine aminotransferase. AB - Alanine aminotransferase (ALT) catalyzes the reversible transamination between alanine and 2-oxoglutarate to form pyruvate and glutamate, and thereby has a key role in the intermediary metabolism of glucose and amino acids. Two ALT isoenzymes are known to exist, but only one ALT gene has been cloned, GPT. In this study, we cloned a homolog of GPT and named it GPT2, and the corresponding protein ALT2. GPT2 shares 69% identity and 78% similarity at the protein level to the previously cloned GPT. The human gene GPT2 encodes a 3.9-kb mRNA, consists of 12 exons, spanning approximately 50 kb of the genome, and maps to chromosome 16q12.1. GPT2 and GPT differ in mRNA expression in that GPT2 is highly expressed in muscle, fat, and kidney, whereas GPT is mainly expressed in kidney, liver, and heart. In addition, GPT2 seems to be the predominant form of GPT at the mRNA level in these tissues. Expression of ALT2 protein in Escherichia coli produced a functional recombinant enzyme that catalyzes alanine transamination, confirming that the enzyme is an ALT. The more abundant expression of GPT2 than GPT, especially in muscle and fat, suggests a unique and previously unrecognized role of this gene product in glucose, amino acid, and fatty acid metabolism and homeostasis. PMID- 11863377 TI - Parkinson's disease and apolipoprotein E: possible association with dementia but not age at onset. AB - Idiopathic Parkinson's disease (PD) is an age-dependent, neurodegenerative condition frequently associated with dementia. Although it is predominantly a sporadic disease, 20-30% of cases are familial, suggesting a complex mode of inheritance. Apolipoprotein E (APOE) allele epsilon4 has been associated with familial and sporadic late-onset senile dementia of the Alzheimer's type. To investigate the role of this gene in the development of dementia associated with PD and age at onset of PD, we evaluated the frequency of APOE gene polymorphism in a sample of PD patients with (n=118) and without (n=167) a family history, as well as matched normal controls (n=96). The PD sample was categorized according to age at onset and presence or absence of dementia. Kaplan-Meier survival analysis was used to plot genotype-specific age at onset distribution curves. Allele frequencies of APOE in PD patients with and without a family history and normal controls were not significantly different. APOE genotypes were also similar between the groups. However, the frequencies of epsilon4 allele and epsilon4/- genotype in the PD group with dementia were more than twofold higher than in normal controls, and the differences were statistically significant. There were no differences in the allele and genotype frequencies of the APOE gene between PD groups with different age at onset. The familial PD had significantly earlier age at onset than sporadic PD (Log-rank test, P=0.027). The age at onset distribution curves for different genotype groups were similar, and their differences were not statistically significant (P=0.38). After the Bonferroni's correction for multiple tests, the positive results are not significant at the P<0.05 level. We conclude that APOE does not play an important role in susceptibility to PD or age at onset of PD, but may play a role in dementia associated with PD in our sample. PMID- 11863378 TI - Seasonal differences in the hormonal control of territorial aggression in free living European stonechats. AB - In birds, territorial aggression during the breeding season is regulated by testosterone (T). However, many bird species also express aggressive behavior during the nonbreeding season, when plasma levels of T are low. It has been suggested that during this period estrogens might play a major role in regulating territorial aggression. In the present study we compared the effects of simultaneous blockage of androgenic and estrogenic actions on territorial aggression during the breeding and nonbreeding seasons in free-living male European stonechats (Saxicola torquata rubicola). European stonechats are of particular interest since they establish territories and form pairs during both the breeding and the nonbreeding seasons. Thus territorial aggression and its endocrine control can be compared between reproductive and non-reproductive contexts. Inhibition of androgenic and estrogenic actions by simultaneous application of Flutamide and ATD reduced territorial aggression during the breeding season, but not during the nonbreeding season. Our results show that androgens and/or estrogens are involved in the endocrine control of territorial aggression in stonechats only in a reproductive context, but not in a non reproductive one. PMID- 11863379 TI - Glucocorticoids and parental hyperphagia in ring doves (Streptopelia risoria). AB - These studies explored the possibility that glucocorticoids promote parental care in ring doves by mediating, at least in part, the pronounced increase in food consumption that parent doves exhibit while provisioning their young. Plasma concentrations of the endogenous glucocorticoid corticosterone were found to be significantly higher in breeding females during the posthatching phase than during the incubation period. These differences were not observed in male breeding partners, but sex differences in daily activity rhythms are well documented in breeding doves, and blood sampling at different times of day would be required to adequately characterize the pattern of corticosterone in males during these breeding stages. In studies on nonbreeding doves, twice-daily intracerebroventricular (icv) injections of the synthetic glucocorticoid dexamethasone (DEX) increased food intake by 25-50% in both sexes, and further studies in males revealed that the increase was directly related to the dose of DEX administered. The highest dose of DEX given icv (1.0 microg/day) was not effective in stimulating feeding when given systemically, thereby suggesting that the hyperphagic action of DEX is exerted directly on the central nervous system. The icv infusion of the selective glucocorticoid receptor antagonist RU38486 blocked the hyperphagic effects of twice-daily icv injections of DEX in both sexes. Collectively, these data support the hypothesis that corticosterone contributes to the parental hyperphagia exhibited by breeding doves during the posthatching period. They also suggest that these orexigenic effects are mediated in part by CNS binding sites that resemble mammalian glucocorticoid receptors. PMID- 11863380 TI - Effects of ovariectomy and estrogen treatment on learning and hippocampal neurotransmitters in mice. AB - This study examined the effects of long-term estrogen treatment (sc 17 beta estradiol minipellets) on learning in C57BL/6J female and male mice using a position discrimination task in the T-maze and a win-stay task (1/8 arms baited) in the radial arm maze (RAM). In addition, hippocampal monoamines and ChAT activity were measured at the end of the study and correlated to task performance. Female sham-operated (gonadally intact) and ovariectomized (OVX) mice were treated with estrogen either for 7 or 40 days before the behavioral tests and intact male mice for 7 days before the behavioral tests. In sham operated mice the 40-day estrogen treatment improved RAM performance and in OVX mice both the 7- and 40-day estrogen treatments improved the performance in both maze tasks. The estrogen treatment also improved RAM performance in males. The hippocampal ChAT, NA, 5-HIAA, and DOPAC levels were decreased in OVX mice. Furthermore, the effects of estrogen treatment on the levels of hippocampal 5-HT and its metabolite 5-HIAA were different in sham-operated than in OVX mice. We could find no correlation between cognitive measures and neurochemical variables. This study gives new information about the effects of estrogen on learning and hippocampal neurotransmitters in mice. PMID- 11863381 TI - Neonatal handling enhances contextual fear conditioning and alters corticosterone stress responses in young rats. AB - Previous studies have indicated that neonatal handling influences development of hypothalamic-pituitary-adrenal (HPA) control of corticosterone. In addition, corticosterone influences memory consolidation processes in contextual fear conditioning. Therefore, neonatal handling may affect hippocampal-dependent memory processes present in contextual fear conditioning by influencing the development of HPA control of corticosterone. To investigate the effects of neonatal handling on early learning, rat pups were either handled (15-min removal from home cage) on the first 15 days after birth or left undisturbed in their home cage. Handled rats and nonhandled rats were fear conditioned at 18, 21, or 30 days of age and then tested at two time points--24 h following conditioning and at postnatal day 45. Subsequently, at approximately postnatal day 60, rats were exposed to restraint stress and corticosterone levels were assessed during restraint and recovery. Handled and nonhandled rats did not differ significantly in their freezing response immediately following footshock on the conditioning day. However, when tested for contextual fear conditioning at 24 h following conditioning and at postnatal day 45, handled rats showed more freezing behavior than nonhandled rats. When exposed to restraint stress, handled rats had a more rapid return of corticosterone to basal levels than nonhandled rats. These results indicate that neonatal handling enhances developmentally early memory processes involved in contextual fear conditioning and confirms previously reported effects of neonatal handling on HPA control of corticosterone. PMID- 11863382 TI - Behavioral thermoregulation and the role of melatonin in a nocturnal snake. AB - Daily and seasonal variations in hormone levels influence the complex interactions between behavior and physiology. Ectothermic animals possess the unique ability behaviorally to adjust body temperature (T(b)) to control physiological rate processes. Thus, a hormone may indirectly influence a physiological rate by directly influencing the behaviors that adjust or control that rate process. Although many hormonal influences on behavioral regulation of T(b) remain uninvestigated, melatonin (MEL) generally is considered a hormone that decreases mean preferred T(b). Many ectotherms demonstrate the selection of lower T(b)'s in response to increased MEL concentrations. Here, we examined the influence of MEL on the behavioral regulation of T(b) in the nocturnal African house snake Lamprophis fuliginosus. A series of experiments with two injection regimes of MEL had no significant effect on the mean preferred T(b) of L. fuliginosus. In addition, mean preferred T(b)'s during the photophase did not differ significantly from those during scotophase. Our findings suggest that L. fuliginosus does not respond to elevated concentrations of either endogenous or exogenous MEL. To verify that the African house snake is nocturnal, we investigated activity patterns of L. fuliginosus throughout the photoperiod. The activity period of L. fuliginosus occurs in the scotophase of the photoperiod, a pattern consistent with that of nocturnal species. This suggests that nocturnal organisms such as L. fuliginosus may not respond to MEL in the same manner as many diurnal species. Our results support the hypothesis that some animals, particularly nocturnal species, may have developed alternative responses to increased plasma concentrations of MEL. PMID- 11863383 TI - Vernal changes in the behavioral and endocrine responses to GnRH application in male European ground squirrels. AB - This field study was aimed at examining hypothalamic involvement in the behavioral changes of male European ground squirrels (Spermophilus citellus) before, during, and after the mating season. The effects of exogenous gonadotropin-releasing hormone (GnRH) application on androgen secretion and behavioral patterns were investigated. Animals were captured, bled, and injected intramuscularly with 40 ng/100 g of GnRH. A second plasma sample was collected 40 min after the treatment to document changes in testosterone secretion. Behavioral parameters such as intra-sexual aggression, scent marking, and home range size were compared on the days before and after the stimulation. In the first two phases, before female emergence and during mating, GnRH-injection caused increases in plasma testosterone. In the post-mating phase, initial plasma testosterone levels had decreased and no elevation could be induced. Sham treatment of controls had no effect in any phase. Conditional parameters like emergence body mass and testicular size covaried with androgen increases only in the pre-mating period. Behavioral changes after GnRH administration occurred during the pre-mating period. Intra-sexual aggression, scent marking, and home range size increased significantly in experimental individuals. Later, during mating and post-mating, we found no behavioral changes associated with the GnRH treatment or the testosterone increase. The results demonstrate changes in the endocrine and behavioral sensitivity to GnRH application, according to the phases of the active season. An exogenous pulse of GnRH can apparently release behavior in male European ground squirrels, which is normally context dependent with the emergence of females. PMID- 11863384 TI - Chronic neuropeptide Y infusion during lactation suppresses pup growth and reduces the length of lactational infertility in rats. AB - In lactating rats, food restriction potentiates the already high levels of hypothalamic neuropeptide Y (NPY). To investigate the role that high levels of NPY might play in the prolongation of lactational infertility that typically accompanies a food restricted lactation we investigated the effects of chronic central infusions of NPY in ad libitum-fed lactating females. First, we compared the effects of intracerebroventricular (icv) infusion of NPY from Days 12-19 postpartum at a dose of 14.4 microg/day with a similar treatment in nonlactating females. In subsequent experiments we examined the effects of NPY infusions into the lateral ventricle at doses of 6 or 20 mug/day or unilaterally into the medial preoptic area at a dose of 1 microg/day from either Days 12-19 or 7-21 postpartum. Effects on food intake; female body weight; and, where appropriate, litter weight and length of lactational diestrus were compared between NPY and vehicle-treated females. As expected NPY infusion produced a robust increase in body weight and food intake in nonlactating females that was accompanied by a suppression of cyclicity. By contrast NPY treatment in lactating rats resulted in a marked decrease in litter growth and an earlier termination of lactational diestrus. PMID- 11863385 TI - Effects of testosterone treatment and season on the frequency of dewlap extensions during male-male interactions in the lizard Anolis sagrei. AB - Lizards of the genus Anolis extend and retract a large and often brightly colored throat fan called a dewlap. The dewlap in most anoles is a sexually dimorphic structure. It is larger in males than females and males use the dewlap more frequently and in more contexts than do females. In the present study we investigated whether plasma testosterone (T) levels and season affect the frequency of dewlap use in male--male interactions in the brown anole, Anolis sagrei. We manipulated plasma T levels by implanting adult, castrated males with pellets that delivered no T, a moderate dose of T, or a high dose of T. In tests with stimulus males, castrated males that were treated with a moderate or with a high dose of T had a significantly higher frequency of dewlap extensions than did castrated males that received no T. However, the frequency of dewlap extensions in castrated males that received the high dose of T did not differ significantly from castrated males that received the moderate dose of T or from non-castrated control males. Males captured during the breeding season and tested in the laboratory had a significantly higher frequency of dewlap extensions than did males captured in the postbreeding season and tested in the same manner. These results suggest that plasma T levels affect how frequently males of A. sagrei extend their dewlaps in male--male interactions and that seasonal changes in male dewlap use may be due to seasonal differences in plasma T levels. PMID- 11863386 TI - Parental responsiveness is feminized after neonatal castration in virgin male prairie voles, but is not masculinized by perinatal testosterone in virgin females. AB - We previously found a large sex difference in the parental responsiveness of adult virgin prairie voles (Microtus ochrogaster) such that most males are spontaneously parental, whereas most females are not. Because this sex difference is independent of the gonadal hormones normally circulating in adult virgin voles, the present study examined whether perinatal hormones influence the development of this sex difference. Males were treated prenatally (via their pregnant dam) with both the androgen receptor blocker flutamide (5 mg/day/dam) and the aromatase inhibitor ATD (1 mg/day/dam), or oil, for the last 2 weeks of gestation. Half of the subjects from each group were castrated on the day of birth and the other half received a sham surgery. As adults, intact males were castrated and all males received a silastic capsule filled with testosterone. Prenatal treatment with flutamide and ATD had no effect on males' behavior toward pups, but neonatal castration significantly reduced the percentage of males acting parentally. In a second experiment, females were exposed to testosterone propionate (TP; 50 microg/day/dam) or oil via their dam during the last 2 weeks of gestation. For the first neonatal week, half of the females from each group were injected with TP (1 mg/day) and the other half oil. As adults, females were ovariectomized and half from each group received a testosterone-filled capsule and the other half received an empty capsule. None of the perinatal TP treatments increased females' parental responsiveness, although females from all groups that received testosterone capsules as adults were highly parental. Therefore, although postnatal testicular hormones are necessary for high parental responsiveness in males, the behavior of females is not influenced by perinatal exposure to testosterone. PMID- 11863388 TI - Physical provocation potentiates aggression in male rats receiving anabolic androgenic steroids. AB - Anabolic androgenic steroids (AAS) have been linked to indiscriminant and unprovoked aggression and violence. We employed a brief tail pinch to examine the effects of different AAS on intermale aggression in gonadally intact male rats in response to a mild physical provocation. Animals received 5 mg/kg testosterone propionate (TP), nandrolone (ND), or stanozolol (ST) 5 days/week. Controls received vehicle injections. After 12 weeks, rats were tested for aggression while treatments continued. Animals were paired with either gonadally intact or castrated opponents and were tested in the subject rat's home cage, the opponents's home cage, and a neutral cage. Aggression was tested during tail pinch of the subject rat and during tail pinch of the opponent rat. In TP-treated males, tail pinch significantly enhanced aggression in all social and environmental conditions compared to intact controls. TP treatment also significantly enhanced aggression when the opponents were tail pinched. Tail pinch did not increase aggression in ND-treated males, and aggression was significantly lower than controls in ST-treated males. As expected, cell nuclear androgen receptor binding was significantly elevated by the high dose of TP. Our results show that while AAS alone does not induce the indiscriminate and unprovoked aggression characteristic of 'roid rage, TP heightens the animals sensitivity to PMID- 11863387 TI - Activation of mu-opioid receptors inhibits lordosis behavior in estrogen and progesterone-primed female rats. AB - The present study investigated the effect of highly selective mu-opioid receptor (OR) agonists on lordosis behavior in ovariectomized rats treated with 3 microg of estradiol benzoate followed 48 h later by 200 microg of progesterone. Ventricular infusion of the endogenous mu-OR agonists endomorphin-1 and -2 suppressed receptive behavior in a time- and dose-dependent fashion. At 6 microg, both endomorphin-1 and -2 inhibited lordosis behavior within 30 min. However, while the effect of endomorphin-1 lasted 60 min, endomorphin-2 inhibition lasted up to 120 min after infusion. Pretreatment with naloxone (5 mg/kg sc) was able to block both endomorphin-1 and endomorphin-2 effects on lordosis. Site-specific infusions of endomorphin-1 or endomorphin-2 into the medial preoptic area (mPOA), the ventromedial nucleus of the hypothalamus (VMH), or into the mesencephalic central gray did not affect receptivity. In contrast, infusion of 1 mug of either compound into the medial septum/horizontal diagonal band of Broca inhibited lordosis in a pattern very similar to that seen after intraventricular infusions. Infusion of the potent synthetic mu-OR agonist [D-Ala(2),N-Me-Phe(4),Gly-ol(5)] enkephalin (0.08 microg) into the VMH and mPOA inhibited lordosis behavior for at least 60 min after infusion. The nonspecific opioid receptor antagonist naloxone was able to facilitate lordosis in partially receptive female rats when infused into the mPOA but not when infused into the VMH. The behavioral effects of the agonists and antagonist used in this study suggest that the endogenous mu-opioid system modulates estrogen and progesterone-induced lordosis behavior. PMID- 11863389 TI - The fireside hypothesis: was there differential selection to tolerate air pollution during human evolution? AB - It is believed that sometime around 1.9 million years ago early hominid ancestors began to migrate out of Africa. Migration north and away from the equator would have forced early humans to encounter seasonal fluctuations in temperature. As a means of adapting to cold climates, the use of fire undoubtedly played an important role. We hypothesize that progressive exposure to toxic airborne particles produced from combustion created selective pressure to develop an adaptation to the inhalation of smoke. In this paper we test this hypothesis using archival data on the incidence of different cancers among geographically distinct human populations. As predicted from evolutionary theory, the incidence of lung cancer is inversely proportional to the ostensible reliance on fire of geographically different groups during human evolutionary history. PMID- 11863390 TI - Ovarian interleukin-1-induced gene expression: privileged genes threshold theory. AB - Interleukin (IL)-1, an established mediator of inflammation, is also a mediator of ovulation (a cyclic inflammatory-like process). We have shown that IL-1 beta induces the in vitro expression of genes believed to play important role in ovulation (IL-1 beta itself, its receptors, IL-1 beta receptor antagonist, glucose transporters 1 and 3, secretory and cytosolic phospholipase A(2), prostaglandin endoperoxide synthase 1 and 2). These experiments suggest that the target genes are turned on over a relatively narrow IL-1 beta dose range. Moreover, IL-1 induces gene expression in what appears to be a hierarchical manner. We hypothesize that IL-1 induces a host of ovulation-associated genes, in a manner that is not only dose-dependent, but also obeys a certain hierarchical order, serving as 'check gates' in securing successful ovulation. PMID- 11863391 TI - The generation of psychosis: a pragmatic approach. AB - The paper discusses the role of speech in the generation of psychosis. The traditional phenomenological approach describes schizophrenic speech as desocialized, autistic and destructive. Based on 'Speech Act Theory', we argue that patients in an acute psychotic state assign maximal illocutionary force to their utterances and mark these speech acts as felicitous. We hypothesize that the pragmatic approach can serve a special role in bilingual patients, the mother tongue being more pronounced in the generation of the psychosis. This view gains support from clinical experience and case studies and can be used as a treatment strategy for bilingual patients. PMID- 11863392 TI - Physiologic and pathologic myocardial hypertrophy--physiologic and pathologic regression of hypertrophy? AB - Hypertrophy of the left ventricle is an adaptive phenomenon of ambiguous biological value. It enables improvement of the heart performance without substantial enhancement of energetic demands. On the other hand, pathologic left ventricular hypertrophy (LVH) is characterized by increased fibrosis, diminished coronary flow reserve and protein remodeling, resulting in increased cardiovascular morbidity and mortality. Achievement of LVH regression is thus considered a principal therapeutic aim. However, the reversal of LVH is a very complex process in which both hemodynamic and non-hemodynamic alterations participate. Reversal of LVH does not mean the re-expression of the original genotype and normalization of myocardial structure and function. It does not guarantee that the heart will be normal in all aspects. Regression of hypertrophy induced by different therapeutic means may exhibit different properties and patterns, with variable biological implications. Physiologic growth stimulators seem to induce LVH without prognostically undesirable alterations. It is a challenge to determine which approach to treatment of hemodynamic overload and concomitant LVH is optimal. PMID- 11863393 TI - Can the sex of the second child be predicted by the birth-weight of the first child? AB - The reproduction costs to a human mother are different if she has a son as opposed to a daughter. According to the Trivers-Willard hypothesis, evolution should promote those females who, having the ability to invest in the more expensive sex, are also able to adjust the sex of their offspring accordingly. It is therefore possible that a mother's biological condition (which is also connected with her reproductive potential), as measured by the neonatal weight of her first child, can be a good predictor of the second child's sex. From data for 227 healthy mothers from Wroclaw (Poland) we show that the probability of giving birth to a boy in the second pregnancy is higher after a relatively heavier first born child (ANOVA, F(1,225)=3.79, P<0.053). This relationship, however, is only significant after a first-born daughter (F(1,117)=9.66, P<0.002) and not after a first-born boy. Some possible explanations of the fact that only the birth-weight of a first-born daughter--and not a son--can be a good predictor of the secondary sex ratio are also discussed. PMID- 11863394 TI - Activation and release of degradative proteinases within the myocardium are the trigger for ventricular remodeling in chronic heart failure. AB - Our conceptual framework of chronic heart failure is based upon the neurohormonal model. In this construct, neurohormonal systems that provide short-term homeostasis remain activated after a myocardial injury, producing progressive ventricular dysfunction and worsening heart failure. However, this model fails to explain several aspects of the pathophysiology of heart failure, including the mechanisms that trigger neurohoromone release and those that lead to ventricular dysfunction in the absence of a large myocardial infarction. These gaps in our understanding can be explained by an expanded model of heart failure, which focuses on myocardial matrix events as the triggers for disease progression. This model embraces the neurohormonal model, and integrates the roles of the immune system and the myocardial fibroblast, within the matrix, to more fully describe the initiation and progression of the disease. PMID- 11863395 TI - Delirium and psychotic symptoms--an integrative model. AB - Delirium may be a common cause of psychotic symptoms such as hallucinations, bizarre delusions and thought-disorder, even in conditions such as schizophrenia, mania and depression, where delirium has traditionally been excluded by definition. This situation is a consequence of the insensitivity of current clinical criteria for the diagnosis of delirium, which recognize only the most severe forms of functional brain impairment (including disorientation and clouding of consciousness). Serial electroencephalograms (EEGs) are the most sensitive method for detecting delirium, and until such studies are performed, the true incidence of delirium in psychotic patients will not be known. The suggested causal mechanism of delirium in psychosis is sleep disruption. Sleep is essential for maintenance of memory circuits, which otherwise suffer progressive synaptic weakening due to molecular turnover. When sleep is disrupted, memory circuits deteriorate, and subsequent activation of incompetent circuits can generate psychotic symptoms. Induction of physiologically normal sleep would therefore be expected to produce significant clinical improvement in patients with psychotic symptoms. Furthermore, the 'anti-delirium' action of electroconvulsive therapy may account for its effectiveness in alleviating a wide range of psychiatric and neurological pathologies. PMID- 11863396 TI - Weight-loss drugs and supplements: are there safer alternatives? AB - Obesity is a major cause of health complaints in western developed countries. Problems ranging from apnea to joint pain have been associated with excess weight. Many factors have been attributed to the epidemic of obesity including sedentary lifestyle, high-fat diets and consumption of large amounts of processed foods. Pharmacies and health-food store shelves abound with a vast selection of products promoted for weight-loss. Some of these have made headlines recently for the damaging effect they have on such things as cardiac valvular function. Unfortunately, others will probably follow and original data is presented on potentially dangerous natural products. Alternatives are presented and discussed below. These natural alternatives include such things as digestive enzyme inhibitors (e.g. L-arabinose, hibiscus tea, marine algae, Nomame Herba, etc), anorexics (e.g. monoterpenes such as d-limonene and perillyl alcohol), glucose uptake inhibitors (e.g. phlorizin), and probiotics as adjuvants. These all natural products are presented as some possible alternatives to those that could be potentially lethal and are not meant as the only options. PMID- 11863399 TI - A common element in the immediate inducement of effortless, natural-sounding, fluent speech in people who stutter: 'the second speech signal'. AB - Recent attempts to find a common element in the inducement of fluent speech have focused on Wingate's notion that fluency can be induced via an altered manner of speaking by placing an emphasis on phonation. The problem with this notion of fluency enhancement is that it appears to be too expansive a scheme of fluency. The schemata allows for any forward flowing speech or speech-like act to be considered fluent despite the increased cognitive mediation, the extra effort involved in implementing the procedure, the unnaturalness of the resultant end product, and a lack of stability over time. It is proposed that the only significant characteristics of 'true fluency' are that it is indistinguishable from the typical fluent speech of people who do not stutter, and that it is effortless in nature. This is achieved via the use of a second speech signal which is not cognitive in nature, is operationally delineated, and is easy to manipulate. PMID- 11863397 TI - Streptomycin revisited: molecular action in the microbial cell. AB - The key event for antimicrobial action begins when streptomycin binds to the 30S subunit (S12 protein) of a ribosome. Lysine 42 and lysine 87 are involved. It is proposed that antagonism of acetyl coenzyme A carboxylase by streptomycin results in faulty fatty acids, lipids and derivatives marked exclusively for cell membrane synthesis. Streptomycin-sensitive growing cells are fatally wounded when defective membranes leak K(+) ions, then amino acids, nucleotides, oligonucleotides and proteins as increasing amounts of streptomycin enter the cell. PMID- 11863398 TI - The central role of magnesium deficiency in Tourette's syndrome: causal relationships between magnesium deficiency, altered biochemical pathways and symptoms relating to Tourette's syndrome and several reported comorbid conditions. AB - Prior studies have suggested a common etiology involved in Tourette's syndrome and several comorbid conditions and symptomatology. Reportedly, current medications used in Tourette's syndrome have intolerable side-effects or are ineffective for many patients. After thoroughly researching the literature, I hypothesize that magnesium deficiency may be the central precipitating event and common pathway for the subsequent biochemical effects on substance P, kynurenine, NMDA receptors, and vitamin B6 that may result in the symptomatology of Tourette's syndrome and several reported comorbid conditions. These comorbid conditions and symptomatology include allergy, asthma, autism, attention deficit hyperactivity disorder, obsessive compulsive disorder, coprolalia, copropraxia, anxiety, depression, restless leg syndrome, migraine, self-injurious behavior, autoimmunity, rage, bruxism, seizure, heart arrhythmia, heightened sensitivity to sensory stimuli, and an exaggerated startle response. Common possible environmental and genetic factors are discussed, as well as biochemical mechanisms. Clinical studies to determine the medical efficacy for a comprehensive magnesium treatment option for Tourette's syndrome need to be conducted to make this relatively safe, low side-effect treatment option available to doctors and their patients. PMID- 11863400 TI - Reduction of stuttering: the dual inhibition hypothesis. AB - Treatment for stuttering attempts to reduce or eliminate the observable core markers of the disorder, specifically repetitions and prolongation. In this hypothesis, it is proposed that stuttering may be inhibited by two distinct yet related procedures: active inhibition and passive inhibition. Active inhibition is brought about when the person who stutters makes volitional changes to his or her speaking pattern, such as when employing behavioral modification techniques. Passive inhibition automatically inhibits the involuntary stuttering block and can be induced from an external source, such as altered auditory feedback, or by the use of sufficient active inhibition. It is suggested that passively inhibiting stuttering results in speech that is more automatic, natural sounding, and truly fluent speech than the speech that is derived primarily from active inhibition. Evidence of passive inhibition resulting from active inhibition can be seen when people who stutter exhibit uncontrolled fluency following behavioral therapy. PMID- 11863401 TI - Meridians in acupuncture and infrared imaging. AB - The meridians in acupuncture are hypothesized to be made up of polarized molecules. Quantum excitations, quasi-particles and others are assumed to be the media of communication between different parts of the body connected by meridians. Infrared pictures are taken to depict the effect of acupuncture on one acupoint of a meridian to a far away pain area. PMID- 11863402 TI - Opioids, sleep, and cancer-related fatigue. AB - Oddly enough, little is known about the effects on sleep of commonly administered analgesic medications. Even less is known about their effect on next-day fatigue, mood, and cognitive functioning. We speculate that part of the fatigue typically experienced by cancer patients can be attributed to disruption of sleep by opioid medications they are taking. Fatigue and sleep are critical to the quality of life of cancer patients. Research is needed to assess the sleep and next-day consequences that can be expected from typical doses of different types of pain medications. PMID- 11863403 TI - New antidepressant drugs that do not cross the blood-brain barrier. AB - Stimulation of neurogenesis in the adult brain (i.e. in the hippocampus) has recently been proposed as a putative mechanism of antidepressant action of drugs. This effect of antidepressants may not be achieved by their primary action on proliferating cells, but may involve the drug-triggered mobilization of trophic factors, such as brain-derived neurotrophic factor (BDNF), glia-derived protein S100 beta, or insulin-like growth factor I (IFG-I). Whereas BDNF and S100 beta are produced in the brain, IGF-I is primarily released from peripheral tissues. Administered peripherally, IGF-I increases hippocampal neurogenesis in the adult rat. Because synthesis and release of IGF-I appear to be stimulated by serotonergic mechanisms, we propose that antidepressants that affect serotonergic mechanisms might be rendered more effective by mobilizing IGF-I. Moreover, we suggest that new antidepressant drugs could be designed that would not enter into the brain but would stimulate peripheral mediators such as IGF-I. PMID- 11863404 TI - Can simvastatin promote tumor growth by inducing angiogenesis similar to VEGF? AB - Recent studies suggest that the HMG-CoA reductase inhibitor simvastatin--similar to vascular endothelial growth factor (VEGF)--may promote angiogenesis by activation of a protein kinase Akt-nitric oxide synthase dependent pathway in endothelial cells, an effect that may be beneficial in the treatment of ischemic heart disease. However, induction of angiogenesis by VEGF contributes importantly to the blood supply of developing tumors and tumor metastases as well. Thus, it can be hypothesized that chronic systemic treatment of elderly patients with a drug that induces angiogenesis by a VEGF-like manner will also promote tumor growth. PMID- 11863405 TI - Simple bone cyst is not a single entity: point of view based on a literature review. AB - Three varieties of simple bone cysts have not attracted a lot of clinical attention: simple bone cysts in non-tubular bones, multiple cysts and epiphyseal cysts. Simple bone cysts occurring in non-tubular bones form about 4-10% of total cases, with a higher age incidence and better prognosis. Epiphyseal simple bone cysts have a higher age incidence, a lower male/female ratio, a very high humerus/femur ratio and higher incidence of tibial location than the classic metaphyseal cysts. Multiple cysts occur in the higher age group and show very high male predominance. In the review of the literature, 15 cysts (35.7%) were in non-tubular bones and six cysts (14.3%) were epiphyseal. They have a better prognosis than the classic metaphyseal ones. Four clinico-anatomic varieties of simple bone cysts could be recognized: classic metaphyseal, non-tubular, epiphyseal, and multiple. Venous obstruction might be the cause of all forms of the disease but the etiologies of venous obstruction causing the last three clinical forms might be different from that causing the classic metaphyseal ones. PMID- 11863408 TI - Looking into the eclipse. PMID- 11863406 TI - A gene therapy institute for NIH? PMID- 11863409 TI - Prematurity in gene therapy. PMID- 11863410 TI - Escherichia coli DNA contamination in AmpliTaq Gold polymerase interferes with TaqMan analysis of lacZ. AB - Real-time PCR is a powerful method for the quantification of gene expression in biological samples. This method uses TaqMan chemistry based on the 5' exonuclease activity of the AmpliTaq Gold DNA polymerase which releases fluorescence from hybridized probes during synthesis of each new PCR product. Many gene therapy studies use lacZ, encoding Escherichia coli beta-galactosidase, as a marker gene. Our results demonstrate that E. coli DNA contamination in AmpliTaq Gold polymerase interferes with TaqMan analysis of lacZ gene expression and decreases sensitivity of the method below the level required for biodistribution and long-term gene expression studies. In biodistribution analyses the contamination can lead to false-negative results by masking low level lacZ expression in target and ectopic tissues, and false-positive results if sufficient controls are not used. We conclude that, to get reliable TaqMan results with lacZ, adequate controls should be included in each run to rule out contamination from AmpliTaq Gold polymerase. PMID- 11863411 TI - Molecular engineering of a two-step transcription amplification (TSTA) system for transgene delivery in prostate cancer. AB - Gene therapy is founded on the concept that tissue-specific promoters can express heterologous genes for molecular imaging or therapeutic applications. The engineering of cell-specific enhancers to improve potency and the development of two-step transcriptional activation (TSTA) approaches have significantly improved the efficacy of transgene expression. Here we combine these technologies to create a robust, titratable, androgen-responsive system targeted to prostate cancer cells. Our "chimeric" TSTA system uses a duplicated variant of the prostate-specific antigen (PSA) gene enhancer to express GAL4 derivatives fused to one, two, or four VP16 activation domains. We targeted the resulting activators to cells with reporter templates bearing one, two, or five GAL4 binding sites upstream of firefly luciferase. We monitored activity via firefly luciferase assays in transfected cell extracts and in live nude mice using a cooled charge-coupled device (CCD) imaging system. In this system, we found that firefly luciferase expression in prostate cancer cells can be varied over an 800 fold range. We also found that a single plasmid bearing the optimized enhancer, GAL4-VP16 derivative, and reporter expressed firefly luciferase at 20-fold higher levels than the cytomegalovirus enhancer. We discuss the implications of this strategy and its application to molecular imaging and therapy. PMID- 11863412 TI - Bilamellar cationic liposomes protect adenovectors from preexisting humoral immune responses. AB - Adenoviral vectors have been widely used for gene therapy, but they are limited both by the presence of a humoral immune response that dramatically decreases the level of transduction after reinjection and by their requirement for target cells to express appropriate receptors such as Coxsackie adenovirus receptor (CAR). To overcome both limits, we encapsulated adenovectors using bilamellar DOTAP:chol liposomes. Electron micrography (EM) showed that these liposomes efficiently encapsulated the vectors, allowing CAR-independent adenovector transduction of otherwise resistant cells. DOTAP:chol-encapsulated adenovectors encoding LacZ or alpha(1)-antitrypsin inhibitor (AAT) were also functionally resistant ex vivo and in vivo to the neutralizing effects of human anti-adenoviral antibodies, unlike other liposomal systems. Hence, bilamellar DOTAP:chol liposomes may be useful for applications using adenovectors in which the target cells lack adenoviral receptors or in which the recipient already has or develops a neutralizing antibody response that would otherwise inactivate readministered vector. PMID- 11863413 TI - Comparison of various envelope proteins for their ability to pseudotype lentiviral vectors and transduce primitive hematopoietic cells from human blood. AB - Substantial effort has been invested in developing methodologies for efficient gene transfer into human, repopulating, hematopoietic stem cells. Oncoretroviral vectors are limited by the lack of nuclear mitosis in quiescent stem cells during ex vivo transduction, whereas the preintegration complex of lentiviral vectors contains nuclear-localizing signals that permit genome integration without mitosis. We have developed a flexible and versatile system for generating lentiviral vector particles and have pseudotyped such particles with amphotropic, ecotropic, feline endogenous virus (RD114) or vesicular stomatitis virus (VSV-G) envelope proteins. Particles of all four types could be concentrated approximately 100-fold by ultracentrifugation or ultrafiltration. RD114 or amphotropic particles were more efficient than VSV-G-pseudotyped particles at transducing human cord blood CD34(+) cells and clonogenic progenitors within that population. Amphotropic particles transduced cytokine-mobilized, human peripheral blood CD34(+) cells capable of establishing hematopoiesis in immunodeficient mice more efficiently than the other two types of particles. We conclude that the use of amphotropic pseudotyped lentiviral vector particles rather than the commonly used VSV-G-pseudotyped particles should be considered in potential applications of lentiviral vectors for gene transfer into this therapeutically relevant target cell population. PMID- 11863414 TI - Robust and efficient regulation of transgene expression in vivo by improved tetracycline-dependent lentiviral vectors. AB - We developed a panel of lentiviral vectors that displayed tetracycline-regulated transgene expression over two orders of magnitude in bulk, non-selected populations of transduced cells in vitro and in vivo. The robust expression and homogeneous response indicated that most transduced vector genomes were transcription competent and responsive to regulation, providing the lentiviral vector with a novel competitive advantage for gene transfer. After ex vivo transduction and transplantation of cord blood CD34+ cells into NOD/SCID mice, reporter gene expression could be switched "on" and "off" in human hematopoietic cells in vivo for prolonged times, proving integration of the regulated expression system into long-term repopulating cells. By vector injection into established tumor grafts, we achieved efficient delivery and quantitative regulation of transgene expression in vivo. By these approaches, gene function studies can now be performed in in vivo models of human hematopoiesis and cancer. In the future, regulated lentiviral vectors will improve the safety and efficacy of gene therapy. PMID- 11863415 TI - Prolongation of transgene expression by coexpression of cytokine response modifier a in rodent liver after adenoviral gene transfer. AB - The short duration of expression of the transgenes is a major barrier to the clinical application of adenovirus-mediated gene therapy for hepatic enzyme deficiencies. Previous reports show that Fas-mediated apoptosis has a pivotal role in the rapid elimination of adenovirus-infected hepatocytes. After considering this result and our recent observation that murine hepatocytes can be protected from Fas-mediated apoptosis by expressing cytokine response modifier A (CrmA) in vivo, we hypothesized that CrmA coexpression could also prevent adenovirus-infected hepatocytes from rapid elimination and that this would make prolonged transgene expression achievable in vivo. To examine this, mice with congenital deficiency of lysosomal beta-glucuronidase (GUSB) were infected with recombinant adenoviruses expressing both CrmA and GUSB, and the duration of transgene expression was evaluated. The serum GUSB activity in the mice injected with a recombinant adenovirus expressing GUSB only became undetectable 60 days after the injection, whereas higher than normal GUSB activity was observed for at least 120 days in mice injected with adenoviruses expressing both GUSB and CrmA. Furthermore, we showed that exogenous CrmA expression could prevent the adenovirus-infected hepatocytes from cell death induced by cytotoxic T lymphocytes in vitro. These observations indicate that transgene expression after administration of E1-deleted adenovirus is prolonged by coexpression of the antiapoptotic protein CrmA. PMID- 11863416 TI - Envelope-targeted retrovirus vectors transduce melanoma xenografts but not spleen or liver. AB - Many cancer gene therapy applications would benefit from the development of targeted vectors that could deliver genes in vivo. We have previously achieved efficient in vitro targeting of retrovirus vectors to melanoma cells by fusion of a single chain antibody recognizing the high-molecular-weight melanoma-associated antigen (HMWMAA), followed by a blocking peptide and a matrix metalloprotease cleavage site, to the amino terminus of the murine leukemia virus amphotropic strain envelope. Here we report that up to 3% of cells within an HMWMAA-positive tumor xenograft were infected following a single injection of targeted vector into the tumor and up to 10% of tumor cells became infected when they were co injected with viral producer cells. No infected cells were detected after delivery of targeted vectors to HMWMAA-negative tumor xenografts. Intraperitoneal injection of amphotropic vectors or producer cells resulted in transduction in spleen and liver, which was not detected when targeted vectors or producer cells were used. Our results demonstrate the feasibility of using targeted retroviral vectors for in vivo gene delivery to tumors and highlight the safety benefits of targeted vectors that do not infect other host tissues. PMID- 11863417 TI - The complement response against an oncolytic virus is species-specific in its activation pathways. AB - A variety of oncolytic viruses (OVs) are being tested in clinical trials for different human cancers. Although the innate immune response is critical as the first line of defense in thwarting viral infection of mammalian cells, little is known of this response in the context of OV therapy of tumors. Investigations of activities against a herpes simplex OV demonstrated that HSV-seronegative sera from rats, mice, and humans efficiently neutralize this OV in vitro. Although this neutralization is due to complement, activation of this innate host defense differs in its pathways among species routinely used in preclinical tumor trials. In rats, both natural immunoglobulins and mannan-binding lectin (MBL) activate complement against the OV, while in mice only MBL is relevant to this activation. However, in humans only natural immunoglobulins play a role in complement activity. Quantitative analyses confirm that in vivo complement depletion facilitates the initial infection of tumors by systemic OVs. Therefore, complement activation against oncolytic HSV vectors proceeds through different pathways in different species. These findings are relevant to preclinical rodent studies of OV therapy and their application to human clinical trials. PMID- 11863418 TI - High levels of transduction of human dendritic cells with optimized SIV vectors. AB - As major antigen-presenting cells and effectors in the maintenance of tolerance, dendritic cells (DCs) are key cells of the immune system and can thus be envisioned to have roles in immunotherapy strategies. We, and others, previously showed that simian immunodeficiency virus (SIV)-derived lentiviral vectors were able to deliver a gene into human differentiated DCs. We describe here the upgrading of the SIV vector system and the improvements of the transduction protocol, which allowed us to transduce more than 90% of human monocyte-derived DCs. We developed new SIV lentiviral vectors carrying SIV splice regulatory elements and either the woodchuck hepatitis virus regulatory element (WPRE) or the murine phosphoglycerate-kinase 1 (PGK) promoter. We show that insertion of the WPRE in the SIV vector is detrimental to gene transfer in DCs, while this sequence increases transgene expression in 293T cells. Using an optimized SIV vector, high levels of transgene expression were obtained in more than 30% of human DCs at a multiplicity of infection (MOI) of 1, and close to 100% using a MOI of 20. VSV-G pseudotyped vectors generated with only gag, pol, tat, and rev helper functions failed to transduce DCs. This defect was completely rescued when the SIV accessory gene vpx was provided in trans in vector-producing cells. Genetically modified DCs were shown to behave as bona fide DCs in both allogenic and autologous mixed leukocyte reactions. These findings allow us to propose an optimal system for efficient and safe DC transduction with improved SIV vectors. PMID- 11863419 TI - Induction of a systemic immune response by a polyvalent melanoma-associated antigen DNA vaccine for prevention and treatment of malignant melanoma. AB - Studies have demonstrated that active-specific immunotherapy has potential for controlling melanoma progression. We developed a polyvalent melanoma gene vaccine using a plasmid vector to deliver the immunogenic human melanoma-associated antigens (MAAs) gp100 and TRP-2. The MAA-containing plasmids were delivered individually in vivo using the hemagglutinating virus Japan (HVJ)-anionic liposome delivery system. C57BL/6 mice were immunized weekly by intramuscular (i.m.) injection or intranasal (i.n.) inoculation for 3 weeks. Although both i.m. and i.n. immunization induced Th1 (T helper) and Th2 cell responses to gp100 and TRP2, the i.m. route induced a better Th1 response. MAA-specific IgG2a, IgG1, and delayed-type hypersensitivity (DTH) responses were induced against both MAAs by i.m. immunization. We assessed the vaccine for its prophylactic and therapeutic effect against the murine B16 F10 melanoma. Animals vaccinated and subsequently challenged with a lethal dose of B16 cells were significantly (P<0.01) protected against tumor progression and had significantly (P<0.01) enhanced survival compared with treatment using control plasmid. We also developed a therapeutic model in which mice were given B16 cells and subsequently immunized with the vaccine or treated with control plasmid. In animals treated with the vaccine, tumor growth was significantly (P<0.01) controlled, and survival was prolonged compared with controls. These studies demonstrate that the polyvalent DNA vaccine induces an effective systemic Th response. PMID- 11863420 TI - Glucocorticoid-regulated VEGF expression in ischemic skeletal muscle. AB - Vascular endothelial growth factor (VEGF) is a potent neovascular inducer. Gene therapeutic delivery of a plasmid DNA encoding VEGF has been shown to impart collateral vessel development in animal models of hindlimb ischemia. Constitutive, long-lived expression of VEGF through gene transfer, however, may result in hypervascularization and/or leaky blood vessels. To that end, the introduction of regulated VEGF gene transfer technology may provide a safer and more controlled therapy for ischemic tissues. We developed a glucocorticoid regulated plasmid vector (pNGVL-hAP/GRE(5)-vegf-pA) for modulating VEGF gene expression. This plasmid possessed five tandem repeats of the glucocorticoid responsive element and adenovirus major-late promoter driving the expression of the VEGF(165) cDNA. Intramuscular delivery of this plasmid to mice, and subsequent treatment with the synthetic glucocorticoid dexamethasone (DEX), led to greatly enhanced VEGF expression. Similar delivery to the gracillis muscle of New Zealand white rabbits that had undergone ligation of their femoral artery to induce ischemia exhibited increased VEGF expression and collateral vessel development only in the presence of DEX. Additionally, reintroduction of DEX at a time point during which initial VEGF transgene levels had subsided resulted in a vigorous reinduction of VEGF transgene expression. This new iteration of VEGF gene delivery provides for fine-tuned angiogenic factor-based therapy for tissues requiring neovascularization. PMID- 11863421 TI - Effect of adenovirus gene transfer vectors on the immunologic functions of mouse dendritic cells. AB - To address the effect of adenovirus (Ad) gene transfer vector transduction on the diverse functions of dendritic cells, we used an Ad vector encoding no transgene (AdNull) to transduce mouse bone-marrow-derived dendritic cells (BMDC). Initial experiments using an Ad vector encoding a marker gene (AdGFP, jellyfish green fluorescent protein) showed that the optimal ratio of infectious Ad particles to each cell was 100, when both transgene expression and resultant BMDC viability were taken into account. Exposure to AdNull resulted in upregulation of both surface activation markers (CD40, MHC class II, B7.1, B7.2, ICAM-1) and IL-12 expression by BMDC. AdNull activation of BMDC was observed in multiple strains of mice. Despite this, AdNull-transduced BMDC displayed only modestly impaired antigen uptake ability, as demonstrated in macropinocytosis and phagocytosis assays, in vitro. However, Ad-modified BMDC migrated to regional lymph nodes five times more efficiently than sham-transduced BMDC in vivo. In addition, Ad transduction significantly enhanced the ability of BMDC to present a model peptide antigen to T-lymphocyte hybridoma cells at low BMDC:T cell ratios. We conclude that Ad modification, in and of itself, induces a state of activation in mouse BMDC. This activation, albeit mild compared with that induced by other stimuli, produces measurable effects of the specific immunological functions of these antigen-presenting cells. PMID- 11863422 TI - Retroviral transduction and engraftment ability of primate hematopoietic progenitor and stem cells transduced under serum-free versus serum-containing conditions. AB - The ability to efficiently transduce hematopoietic stem and progenitor cells under serum-free conditions would be desirable for safety and standardization of clinical gene therapy protocols. Using rhesus macaques, we studied the transduction efficiency and engraftment ability of CD34-enriched SCF/G-CSF mobilized progenitor cells (PBSC) transduced with standard amphotropic marking vectors under serum-free and serum-containing conditions. Supernatants were collected from producer cells 16 hours after serum-free medium or medium containing 10% fetal calf serum was added. Vector titers were approximately two- to threefold higher when producer cells were cultured in serum-containing medium. However, retroviral transduction of rhesus CFU-GM was improved using serum-free vector-containing medium. For analysis of engraftment with transduced cells, three macaques had CD34+ peripheral blood stem cells split into two fractions for transduction. One fraction was transduced using serum-free vector-containing medium, and the other fraction was transduced using standard serum-containing medium. The two fractions were re-infused simultaneously following total body irradiation. In all three animals, there was equivalent marking from both vectors for 7-9 months post-transplantation. These data are encouraging regarding the removal of serum-containing medium from clinical hematopoietic cell transduction protocols, given the lack of a detrimental effect on transduction and engraftment with transduced cells. PMID- 11863423 TI - A syringe electrode device for simultaneous injection of DNA and electrotransfer. AB - Electroporation for gene delivery has attracted considerable attention recently, because of both the site-specific nature of the delivery and the high efficiency of the method. Electrotransfer of genes involves the application of an electric field to cells to enhance their permeability, facilitating exogenous polynucleotide transit across the cytoplasmic membrane. However, the relatively high electric field strength required for electroporation induces tissue damage, thus limiting its widespread application. We describe here a syringe electrode of our design, with which the same transfection efficiency can be achieved by using much lower electric field strength than that of conventional electrodes, such that the tissue damage is minimized. PMID- 11863424 TI - Altering retroviral tropism using a random-display envelope library. AB - Tissue-specific gene delivery is an important aspect of many gene therapy applications. The experiments reported here constitute the first successful demonstration that cell-specific entry can be obtained by screening a random library of retroviral envelope proteins produced from a mammalian cell system. The library consisted of 10(6) different subgroup A feline leukemia virus envelope protein variants with 10 randomly substituted amino acids in the receptor-determining region. Selecting the library for fully functional envelope proteins able to mediate stable gene transfer resulted in the identification of a single envelope protein variant (EF). Subsequent examination of the host range of EF revealed that it was highly specific for D17 canine osteosarcoma cells. This was in contrast to the host ranges of the parental subgroup A and closely related subgroup C envelope proteins. Interference assays on D17 cells further indicated that receptor usage by EF was also altered compared with the A and C envelope proteins. The EF envelope protein thus isolated should be useful for studying gene therapy treatments of osteosarcoma in a large-animal model. PMID- 11863426 TI - Structure and function of voltage-dependent ion channel regulatory beta subunits. AB - Voltage-dependent K(+), Ca(2+), and Na(+) channels play vital roles in basic physiological processes, including management of the action potential, signal transduction, and secretion. They share the common function of passively transporting ions across cell membranes; thus, it would not be surprising if they should exhibit similarities of both structure and mechanism. Indeed, the principal pore-forming (alpha) subunits of each show either exact or approximate 4-fold symmetry and share a similar transmembrane topology, and all are gated by changes in membrane potential. Furthermore, these channels all possess an auxiliary polypeptide, designated the beta subunit, which plays an important role in their regulation. Despite considerable functional semblences and abilities to interact with structurally similar alpha subunits, however, there is considerable structural diversity among the beta subunits. In this review, we discuss the similarities and differences in the structures and functions of the beta subunits of the voltage-dependent K(+), Ca(2+), and Na(+) channels. PMID- 11863427 TI - The function of photosystem I. Quantum chemical insight into the role of tryptophan-quinone interactions. AB - Quantum chemical calculations have provided evidence for the role of tryptophan residues in the electron transfer process of photosystem I (PS-I). The interaction of Trp with quinone acceptors and their radical anions in the A(1) site of PS-I has been modeled by various indole-quinone and indole-semiquinone complexes. MP2 optimizations show that, while neutral quinones and an indole molecule prefer a pi-stacked arrangement, semiquinone radical anions prefer a T stacked conformation with significant N-H...pi hydrogen bonding interactions. Comparison of density functional calculations of electronic g-tensors with electron paramagnetic resonance data strongly suggests that hydrogen-bonded T shaped arrangements occur upon reduction of quinone acceptors without an extended side chain (e.g., duroquinone or naphthoquinone), when reconstituted into the phylloquinone-depleted A(1) site of PS-I. In contrast, for the native phylloquinone (vitamin K(1), Q(K)), reorientation of the semiquinone radical anion is prevented by side chain-protein interactions. For a fixed pi-stacked arrangement, the extent of the intermolecular interaction is reduced upon one electron reduction. This corresponds to a lowering of the redox potential of the P(700)(+)*Q(K)(-)* radical pair, due to interactions of Q(K) with a tryptophan. Together with the comparably weak hydrogen bonding in PS-I, the proposed model explains the very negative redox potential of the A(1) site, needed for forward electron transfer. T-stacking hydrogen bonds to semiquinones may also have to be considered in many other electron transfer processes in living organisms. PMID- 11863428 TI - A minimal exonuclease domain of WRN forms a hexamer on DNA and possesses both 3'- 5' exonuclease and 5'-protruding strand endonuclease activities. AB - Werner syndrome is a rare autosomal recessive disease characterized by a premature aging phenotype, genomic instability, and a dramatically increased incidence of cancer and heart disease. Mutations in a single gene encoding a 1432 amino acid helicase/exonuclease (hWRN) have been shown to be responsible for the development of this disease. We have cloned, overexpressed, and purified a minimal, 171-amino acid fragment of hWRN that functions as an exonuclease. This fragment, encompassing residues 70-240 of hWRN (hWRN-N(70-240)), exhibits the same level of 3'-5' exonuclease activity as the previously described exonuclease fragment encompassing residues 1-333 of the full-length protein. The fragment also contains a 5'-protruding DNA strand endonuclease activity at a single-strand double-strand DNA junction and within single-stranded DNA, as well as a 3'-5' exonuclease activity on single-stranded DNA. We find hWRN-N(70-240) is in a trimer-hexamer equilibrium in the absence of DNA when examined by gel filtration chromatography and atomic force microscopy. Upon addition of DNA substrate, hWRN N(70-240) forms a hexamer and interacts with the recessed 3'-end of the DNA. Moreover, we find that the interaction of hWRN-N(70-240) with the replication protein PCNA also causes this minimal, 171-amino acid exonuclease region to form a hexamer. Thus, the active form of this minimal exonuclease fragment of human WRN appears to be a hexamer. The implications these results have on our understanding of hWRN's roles in DNA replication and repair are discussed. PMID- 11863429 TI - Direct detection of hydrogen bonds in monomeric superoxide dismutase: biological implications. AB - Hydrogen bonds were directly determined via NMR with different experimental approaches at 600 and 800 MHz for reduced monomeric superoxide dismutase (Q133M2SOD, 16 kDa). This protein contains a copper and a zinc ion and shows the classical superoxide dismutase (SOD) eight-stranded beta-barrel fold. The best results for this intermediate molecular mass protein were obtained using a TROSY version of the long-range HNCO experiment at high magnetic field (800 MHz) or with a cryoprobe at 600 MHz. The backbone hydrogen bond network that defines the secondary structure of the protein was detected. Thirty-five backbone hydrogen bonds were identified. The lower limit for their detection, their relation to the TROSY R(2) rates, and the correlation between hydrogen bond detectability and signal line width are discussed. Experiments were also optimized to detect hydrogen bonds involving key side chains, which lead to the observation of five hydrogen bonds. In particular, the hydrogen bonds involving the side chain of Asp 124 were observed, which show significant differences with respect to the bonds expected on the basis of the crystal structure. The relevance of this finding relies also on the fact that Asp 124 is a key residue in determining the affinity of the protein for zinc. It has now been determined that the gain of the toxic function of peroxynitrite formation in SOD mutants related to amyotrophic lateral sclerosis (ALS) is due to SOD species lacking the zinc ion, as a consequence of a reduced affinity for zinc. Therefore, this study provides structural hints for understanding the origin of the enzymatic behavior of the Zn-deficient SOD. PMID- 11863430 TI - Structure and spectroscopy of the periplasmic cytochrome c nitrite reductase from Escherichia coli. AB - The crystal structure and spectroscopic properties of the periplasmic penta-heme cytochrome c nitrite reductase (NrfA) of Escherichia coli are presented. The structure is the first for a member of the NrfA subgroup that utilize a soluble penta-heme cytochrome, NrfB, as a redox partner. Comparison to the structures of Wolinella succinogenes NrfA and Sulfospirillum deleyianum NrfA, which accept electrons from a membrane-anchored tetra-heme cytochrome (NrfH), reveals notable differences in the protein surface around heme 2, which may be the docking site for the redox partner. The structure shows that four of the NrfA hemes (hemes 2 5) have bis-histidine axial heme-Fe ligation. The catalytic heme-Fe (heme 1) has a lysine distal ligand and an oxygen atom proximal ligand. Analysis of NrfA in solution by magnetic circular dichroism (MCD) suggested that the oxygen ligand arose from water. Electron paramagnetic resonance (EPR) spectra were collected from electrochemically poised NrfA samples. Broad perpendicular mode signals at g similar 10.8 and 3.5, characteristic of weakly spin-coupled S = 5/2, S = 1/2 paramagnets, titrated with E(m) = -107 mV. A possible origin for these are the active site Lys-OH(2) coordinated heme (heme 1) and a nearby bis-His coordinated heme (heme 3). A rhombic heme Fe(III) EPR signal at g(z) = 2.91, g(y) = 2.3, g(x) = 1.5 titrated with E(m) = -37 mV and is likely to arise from bis-His coordinated heme (heme 2) in which the interplanar angle of the imidazole rings is 21.2. The final two bis-His coordinated hemes (hemes 4 and 5) have imidazole interplanar angles of 64.4 and 71.8. Either, or both, of these hemes could give rise to a "Large g max" EPR signal at g(z)() = 3.17 that titrated at potentials between 250 and -400 mV. Previous spectroscopic studies on NrfA from a number of bacterial species are considered in the light of the structure-based spectro potentiometric analysis presented for the E. coli NrfA. PMID- 11863431 TI - Identification of a thiosulfate utilization gene cluster from the green phototrophic bacterium Chlorobium limicola. AB - Chlorobium is an autotrophic, green phototrophic bacterium which uses reduced sulfur compounds to fix carbon dioxide in the light. The pathways for the oxidation of sulfide, sulfur, and thiosulfate have not been characterized with certainty for any species of bacteria. However, soluble cytochrome c-551 and flavocytochrome c (FCSD) have previously been implicated in the oxidation of thiosulfate and sulfide on the basis of enzyme assays in Chlorobium. We have now made a number of observations relating to the oxidation of reduced sulfur compounds. (1) Western analysis shows that soluble cytochrome c-551 in Chlorobium limicola is regulated by thiosulfate, consistent with a role in the utilization of thiosulfate. (2) A membrane-bound flavocytochrome c-sulfide dehydrogenase (which is normally a soluble protein in other species) is constitutive and not regulated by sulfide as expected for an obligately autotrophic species dependent upon sulfide. (3) We have cloned the cytochrome c-551 gene from C. limicola and have found seven other genes, which are also presumably involved in sulfur metabolism and located near that for cytochrome c-551 (SoxA). These include genes for a flavocytochrome c flavoprotein homologue (SoxF2), a nucleotidase homologue (SoxB), four small proteins (including SoxX, SoxY, and SoxZ), and a thiol disulfide interchange protein homologue (SoxW). (4) We have established that the constitutively expressed FCSD genes (soxEF1) are located elsewhere in the genome. (5) Through a database search, we have found that the eight thiosulfate utilization genes are clustered in the same order in the Chlorobium tepidum genome (www.tigr.org). Similar thiosulfate utilization gene clusters occur in at least six other bacterial species but may additionally include genes for rhodanese and sulfite dehydrogenase. PMID- 11863432 TI - Ability of E. coli cyclic AMP receptor protein to differentiate cyclic nucelotides: effects of single site mutations. AB - Escherichia coli cyclic AMP receptor protein (CRP) is a global transcriptional regulator which controls the expression of many different genes. Although different cyclic nucleotides can bind to CRP with almost equal affinity, only in the presence of cAMP could wild-type CRP bind to specific DNA sequences. Molecular genetic studies have identified a class of mutants, CRP*, which either do not require exogenous cAMP for activation or can be activated by cGMP. Thus, these mutants might aid in identifying the structural elements that are involved in the modulation of CRP to correctly differentiate the messages embedded in cyclic nucleotides. In this in vitro study, five CRP* mutants, namely, D53H, S62F, G141Q, G141K, and L148R, were tested for their abilities to bind the lac promoter sequence and the effects of cyclic nucleotides in modulating DNA sequence recognition. For comparison, non-CRP* mutants K52N, T127L, H159L, and K52N/H159L were studied. cCMP and cGMP can replace cAMP as an allosteric effector in all of these CRP mutants except S62F and non-CRP* mutants. The D53H, G141Q, G141K, and L148R mutants exhibit significantly higher affinity for the lac promoter sequence than wild-type CRP while S62F and the non-CRP* mutants exhibit reduced affinity. To probe the pathway of communication, the energetics of subunit assembly in these mutants were monitored by sedimentation equilibrium, and the conformational states of these mutants were probed by proteolysis and accessibility of Cys178 to chemical modifications. Results from these studies imply that signals due to mutations are mostly transmitted through the subunit interface. Thus, residues in CRP outside of the cyclic nucleotide binding site modulate the ability of CRP to differentiate these three cyclic nucleotides through long-range communication. Furthermore, this study shows that CRP* mutations do not impart any unique properties to CRP except that the DNA binding constants are shifted to a regime of higher affinity. PMID- 11863433 TI - Hydrophobicity maps of the N-peptide coiled coil of HIV-1 gp41. AB - Blocking HIV-1 viral entry into the host cell offers a promising new strategy for interfering with the HIV-1 life cycle. A major target of inhibitor design is to prevent binding of fusogenic gp41 C-peptides to the trimeric coiled coil of fusion-active N-peptides. Here, we map the hydrophobic character of the binding surface of the IQN17 peptide, a soluble analogue of the N-peptide coiled coil. The local binding affinity for a hydrophobic probe is determined by three methods: a hydrophobic force field, and molecular dynamics in solution analyzed by test particle insertion and inhomogeneous information theory. The regions of highest calculated hydrophobicity overlap with the positions of the hydrophobic anchor residues of the native C-peptides, and of two known inhibitors. Additional binding sites not exploited by these inhibitors are identified, and modifications for enhancing their binding affinity are suggested. PMID- 11863434 TI - A nonhydrolyzable reactive cAMP analogue, (S(p))-8-[(4-bromo-2,3 dioxobutyl)thio]adenosine 3',5'-cyclic S-(methyl)monophosphorothioate, irreversibly inactivates human platelet cGMP-inhibited cAMP phosphodiesterase at micromolar concentrations. AB - We previously showed that 8-[(4-bromo-2,3-dioxobutyl)thio]adenosine 3',5'-cyclic monophosphate inactivates cAMP phosphodiesterase (PDE3A); however, millimolar concentrations were needed to inactivate PDE3A because of ongoing hydrolysis. We have now synthesized a nonhydrolyzable reactive cAMP analogue, (S(p))-8-[(4-bromo 2,3-dioxobutyl)thio]adenosine 3',5'-cyclic S-(methyl)monophosphorothioate (S(p)-8 BDB-TcAMPSMe). S(p)-8-BDB-TcAMPSMe inactivates PDE3A in a time-dependent, irreversible manner, exhibiting saturation kinetics with a k(max) of (19.5 +/- 0.3) x 10(-3) min(-1) and a K(I) of 3.5 +/- 0.3 muM. To ascertain whether S(p)-8 BDB-TcAMPSMe reacts in the active site, nonhydrolyzable analogues of the substrate cAMP, or the competitive inhibitor cGMP, were included to protect against the inactivation of PDE3A. The order of effectiveness of protectants in decreasing the rate of inactivation (with K(d) values in micromolar) is as follows: S(p)-cAMPS (18) > R(p)-cGMPS (560) and S(p)-cGMPS (1260) > 5'-AMP (17 660), R(p)-cAMPS (30 110), and 5'-GMP (42 170). We docked S(p)-8-BDB-TcAMPSMe into PDE3A, based on the structural model of PDE3A-cAMP and the kinetic data from site-directed mutants. The S(p)-8-BDB-TcAMPSMe fits into the active site in the model. These results suggest that inactivation of PDE3A by the affinity reagent is a consequence of reaction at the overlap between cAMP and cGMP binding regions in the active site. S(p)-8-BDB-TcAMPSMe has proven to be an effective active site directed irreversible cAMP affinity label for platelet PDE3A and can be used to identify amino acids in the active site of PDE3A as well as in other cAMP phosphodiesterases. PMID- 11863436 TI - The crystal structure of HpcE, a bifunctional decarboxylase/isomerase with a multifunctional fold. AB - The structure of the bifunctional enzyme HpcE (OPET decarboxylase/HHDD isomerase) from Escherichia coli shows that the protein consists of highly similar N and C terminal halves. Sequence matches suggest that this fold is widespread among different species, including man. Many of these homologues are uncharacterized but apparently connected with the metabolism of aromatic compounds. The domain shows similar topology to the C terminal domain of fumarylacetoacetate hydrolase (FAH), a functionally related enzyme, despite lacking significant overall sequence similarity. HpcE is known to catalyze two rather different reactions, and comparisons with FAH allow some tentative conclusions to be drawn about the active sites. Key mutations within the active site apparently allow enzymes with this fold to carry out a variety chemical processes. PMID- 11863435 TI - 3D structure of Torpedo californica acetylcholinesterase complexed with huprine X at 2.1 A resolution: kinetic and molecular dynamic correlates. AB - Huprine X is a novel acetylcholinesterase (AChE) inhibitor, with one of the highest affinities reported for a reversible inhibitor. It is a synthetic hybrid that contains the 4-aminoquinoline substructure of one anti-Alzheimer drug, tacrine, and a carbobicyclic moiety resembling that of another AChE inhibitor, ( )-huperzine A. Cocrystallization of huprine X with Torpedo californica AChE yielded crystals whose 3D structure was determined to 2.1 A resolution. The inhibitor binds to the anionic site and also hinders access to the esteratic site. Its aromatic portion occupies the same binding site as tacrine, stacking between the aromatic rings of Trp84 and Phe330, whereas the carbobicyclic unit occupies the same binding pocket as (-)-huperzine A. Its chlorine substituent was found to lie in a hydrophobic pocket interacting with rings of the aromatic residues Trp432 and Phe330 and with the methyl groups of Met436 and Ile439. Steady-state inhibition data show that huprine X binds to human AChE and Torpedo AChE 28- and 54-fold, respectively, more tightly than tacrine. This difference stems from the fact that the aminoquinoline moiety of huprine X makes interactions similar to those made by tacrine, but additional bonds to the enzyme are made by the huperzine-like substructure and the chlorine atom. Furthermore, both tacrine and huprine X bind more tightly to Torpedo than to human AChE, suggesting that their quinoline substructures interact better with Phe330 than with Tyr337, the corresponding residue in the human AChE structure. Both (-) huperzine A and huprine X display slow binding properties, but only binding of the former causes a peptide flip of Gly117. PMID- 11863437 TI - Inhibition of prostate-specific antigen (PSA) by alpha(1)-antichymotrypsin: salt dependent activation mediated by a conformational change. AB - Prostate-specific antigen (PSA) and its SDS-stable complex with the serine proteinase inhibitor (serpin) alpha(1)-antichymotrypsin (ACT), which is the dominant form of PSA in serum, are in widespread use as markers for the diagnosis of prostate cancer, and there is increasing evidence for the involvement of PSA proteinase activity itself in the development of prostate and other cancers. However, both the formation and degradation of the PSA-ACT complex, denoted PSA*ACT* to indicate substantial changes in the structure of both proteins on complex formation, have been incompletely studied. Here we determine rate and equilibrium constants for the steps involved in PSA*ACT* formation and demonstrate that (a) the effects of added NaCl, polyamines, and Zn(2+) on this process parallel their effects on PSA catalytic activity [Hsieh, M.-C., and Cooperman, B. S. (2000) Biochim. Biophys. Acta 1481, 75-87], (b) the effect of added NaCl in dramatically increasing the rate of ACT inhibition of PSA correlates with salt-induced changes in PSA conformation, and (c) the PSA*ACT* complex is subject to proteolysis by human neutrophil elastase. Possible clinical implications of these findings are considered. PMID- 11863438 TI - Salt bridges destabilize a leucine zipper designed for maximized ion pairing between helices. AB - Interhelical salt bridges are common in leucine zippers and are thought to stabilize the coiled coil conformation. Here we present a detailed thermodynamic investigation of the designed, disulfide-linked leucine zipper AB(SS) whose high resolution NMR structure shows six interhelical ion pairs between heptad positions g of one helix and e' of the other helix but no ion pairing within single helices. The average pK(a) value of the Glu side chain carboxyl groups of AB(SS) is slightly higher than the pK(a) of a freely accessible Glu in an unfolded peptide [Marti, D. N., Jelesarov, I., and Bosshard, H. R. (2000) Biochemistry 39, 12804-12818]. This indicates that the salt bridges are destabilizing, a prediction we now have confirmed by determining the pH +/- stability profile of AB(SS). Circular dichroism-monitored unfolding by urea and by heating and differential scanning calorimetry show that the coiled coil conformation is approximately 5 kJ/mol more stable when salt bridges are broken by protonation of the carboxyl side chains. Using guanidinium chloride as the denaturant, the increase in the free energy of unfolding on protonation of the carboxyl side chains is larger, approximately 17 kJ/mol. The discrepancy between urea and guanidinium chloride unfolding can be ascribed to the ionic nature of guanidinium chloride, which screens charge-charge interactions. This work demonstrates the difficulty of predicting the energetic contribution of salt bridges from structural data alone even in a case where the ion pairs are seen in high-resolution NMR structures. The reason is that the contribution to stability results from a fine balance between energetically favorable Coulombic attractions and unfavorable desolvation of charges and conformational constraints of the residues involved in ion pairing. The apparent discrepancy between the results presented here and mutational studies indicating stabilization by salt bridges is discussed and resolved. An explanation is proposed for why interhelical salt bridges are frequently found in natural coiled coils despite evidence that they do not directly contribute to stability. PMID- 11863439 TI - Structural basis for the recognition of a bisphosphorylated MAP kinase peptide by human VHR protein Phosphatase. AB - Human VHR (vaccinia H1 related phosphatase) is a member of the dual-specificity phosphatases (DSPs) that often act on bisphosphorylated protein substrates. Unlike most DSPs, VHR displays a strong preference for dephosphorylating phosphotyrosine residues over phosphothreonine residues. Here we describe the 2.75 A crystal structure of the C124S inactive VHR mutant in complex with a bisphosphorylated peptide corresponding to the MAP kinase activation lip. This structure and subsequent biochemical studies revealed the basis for the strong preference for hydrolyzing phosphotyrosine within bisphosphorylated substrates containing -pTXpY-. In the structure, the two phospho residues are oriented into distinct pockets; the phosphotyrosine is bound in the exposed yet deep active site cleft while the phosphothreonine is loosely tethered into a nearby basic pocket containing Arg(158). As this structure is the first substrate-enzyme complex reported for the DSP family of enzymes, these results provide the first glimpse into how DSPs bind their protein substrates. PMID- 11863440 TI - Structure of FAD-bound L-aspartate oxidase: insight into substrate specificity and catalysis. AB - L-Aspartate oxidase (Laspo) catalyzes the conversion of L-Asp to iminoaspartate, the first step in the de novo biosynthesis of NAD(+). This bacterial pathway represents a potential drug target since it is absent in mammals. The Laspo R386L mutant was crystallized in the FAD-bound catalytically competent form and its three-dimensional structure determined at 2.5 A resolution in both the native state and in complex with succinate. Comparison of the R386L holoprotein with the wild-type apoenzyme [Mattevi, A., Tedeschi, G., Bacchella, L., Coda, A., Negri, A., and Ronchi, S. (1999) Structure 7, 745-756] reveals that cofactor incorporation leads to the ordering of two polypeptide segments (residues 44-53 and 104-141) and to a 27 degree rotation of the capping domain. This motion results in the formation of the active site cavity, located at the interface between the capping domain and the FAD-binding domain. The structure of the succinate complex indicates that the cavity surface is decorated by two clusters of H-bond donors that anchor the ligand carboxylates. Moreover, Glu121, which is strictly conserved among Laspo sequences, is positioned to interact with the L Asp alpha-amino group. The architecture of the active site of the Laspo holoenzyme is remarkably similar to that of respiratory fumarate reductases, providing strong evidence for a common mechanism of catalysis in Laspo and flavoproteins of the succinate dehydrogenase/fumarate reductase family. This implies that Laspo is mechanistically distinct from other flavin-dependent amino acid oxidases, such as the prototypical D-amino acid oxidase. PMID- 11863441 TI - Site-directed rotational resonance solid-state NMR distance measurements probe structure and mechanism in the transmembrane domain of the serine bacterial chemoreceptor. AB - The serine receptor of bacterial chemotaxis is an ideal system in which to investigate the molecular mechanism of transmembrane signaling. Solid-state nuclear magnetic resonance (NMR) techniques such as rotational resonance provide a means for measuring local structure and ligand-induced structural changes in intact membrane proteins bound to native membrane vesicles. A general site directed biosynthetic (13)C labeling strategy is used to direct the distance measurements to a specific site; the distance is measured between a unique Cys residue and a non-unique, low-abundance residue (Tyr or Phe). A (13)C-(13)C internuclear distance measurement from (13)CO(i) to (13)C beta(i + 3) at the periplasmic edge of the second membrane-spanning helix (TM2) of 5.1 +/- 0.2 A is consistent with the predicted alpha-helical structure and thus demonstrates an accurate long-distance rotational resonance measurement in the 120 kDa membrane bound receptor. These measurements require a correction for the rotational resonance exchange between the multiple labels of the non-unique amino acid and the natural-abundance (13)C, which is critical to distance measurements in complex systems. A second (13)C-(13)C distance measurement between the transmembrane helices provides a high-resolution measurement of tertiary structure in the transmembrane region. The measured 5.0-5.3 A distance in the presence and absence of ligand is consistent with structural models for the transmembrane region and a proposed signaling mechanism in which ligand binding induces a 1.6 A translation of TM2. This approach can be used for additional measurements of the structure of the transmembrane region and to determine whether the ligand-induced motion is indeed propagated through the transmembrane helices. PMID- 11863442 TI - Themodynamic and transport properties of intermediate states of the photocyclic reaction of photoactive yellow protein. AB - Themodynamic and transport properties of intermediate states of the photocyclic reaction of photoactive yellow protein (PYP) were studied by a combination of the pulsed laser-induced transient grating (TG), transient lens (TrL), and photoacoustic (PA) spectroscopies from tens of nanoseconds to hundreds of milliseconds. The diffusion coefficients (D) of PYP in the ground state (pG) and of the second intermediate state (pB) were determined by the TG analysis, and it was found that D of pG is about 1.2 times larger than D of pB. At the same time, D at various denatured conditions were measured using guanidine hydrochloride as the denaturant. D of completely unfolded protein is about 0.4 times that of the native form. The enthalpy of pB is estimated to be 60 kJ/mol by the TrL method with an assumption that the volume change of pB is not sensitive to the temperature. Since the enthalpy of the first intermediate state (pR) is as high as 160 kJ/mol, it implies that most of the photon energy is stored as the strain of the protein in pR, and this may be the driving force for the successive reaction to pB. From the temperature dependence of the volume change, the difference in the thermal expansion coefficients between pG and pR was calculated. All of the characteristic features of PYP, the negative volume change, the larger thermal expansion coefficient, and the slower diffusion process, indicate that the intermediate pR and pB are reasonably interpreted in terms of the unfolded (loosened) protein structure. PMID- 11863443 TI - Spectral substructure and excitonic interactions in the minor photosystem II antenna complex CP29 as revealed by nonlinear polarization spectroscopy in the frequency domain. AB - CP29 (the lhcb4 gene product), a minor photosystem II antenna complex, binds six chlorophyll (Chl) a, two Chl b, and two to three xanthophyll molecules. The Chl a/b Q(y) absorption band substructure of CP29 (purified from spinach) was investigated by nonlinear polarization spectroscopy in the frequency domain (NLPF) at room temperature. A set of NLPF spectra was obtained at 11 probe wavelengths. Seven probe wavelengths were located in the Q(y) spectral region (between 630 and 690 nm) and four in the Soret band (between 450 and 485 nm). Evaluation of the experimental data within the framework of global analysis leads to the following conclusions: (i) The dominant Chl a absorption (with a maximum at 674 nm) splits into (at least) three subbands (centered at 660, 670, and 681.5 nm). (ii) In the Chl b region two subbands can be identified with maxima located at 640 and 646 nm. (iii) The lowest energy Q(y) transition (peaking at 681.5 nm) is assigned to a Chl a which only weakly interacts with other Chl aor b molecules by incoherent Forster-type excitation energy transfer. (iv) Pronounced excitonic interaction exists between certain Chl a and Chl b molecules, which most likely form a Chl a/b heterodimer. The subbands centered at 640 and 670 nm constitute a strongly coupled Chl a/b pair. The findings of the study indicate that the currently favored view of spectral heterogeneity in CP29 being due essentially to pigment-protein interactions has to be revised. PMID- 11863444 TI - Reduction of the Mn cluster of the water-oxidizing enzyme by nitric oxide: formation of an S(-2) state. AB - The manganese cluster of the oxygen-evolving enzyme of photosystem II is chemically reduced upon interaction with nitric oxide at -30 degrees C. The state formed gives rise to an S = 1/2 multiline EPR signal [Goussias, Ch., Ioannidis, N., and Petrouleas, V. (1997) Biochemistry 36, 9261] that is attributed to a Mn(II)- Mn(III) dimer [Sarrou, J., Ioannidis, N., Deligiannakis, Y., and Petrouleas, V. (1998) Biochemistry 37, 3581]. In this work, we sought to establish whether the state could be assigned to a specific, reduced S state by using flash oxymetry, chlorophyll a fluorescence, and electron paramagnetic resonance spectroscopy. With the Joliot-type O(2) electrode, the first maximum of oxygen evolution was observed on the sixth or seventh flash. Three saturating pre flashes were required to convert the flash pattern characteristic of NO-reduced samples to that of the untreated control (i.e., O(2) evolution maximum on the third flash). Measurements of the S state-dependent level of chlorophyll fluorescence in NO-treated PSII showed a three-flash downshift compared to untreated controls. In the EPR study, the maximum S(2) multi-line EPR signal was observed after the fourth flash. The results from all three methods are consistent with the Mn cluster being in a redox state corresponding to an S(-2) state in a majority of centers after treatment with NO. We were unable to generate the Mn(II)-Mn(III) multi-line signal using hydrazine as a reductant; it appears that the valence distribution and possibly the structure of the Mn cluster in the S(-2) state are dependent on the nature of the reductant that is used. PMID- 11863445 TI - Novel FMN-containing rotenone-insensitive NADH dehydrogenase from Trypanosoma brucei mitochondria: isolation and characterization. AB - A rotenone-insensitive NADH dehydrogenase has been isolated from the mitochondria of the procyclic form of African parasite, Trypanosoma brucei. The active form of the purified enzyme appears to be a dimer consisting of two 33-kDa subunits with noncovalently bound FMN as a cofactor. Hypotonic treatment of intact mitochondria revealed that the NADH dehydrogenase is located in the inner membrane/matrix fraction facing the matrix. The treatment of mitochondria with increasing concentrations of digitonin suggested that the NADH dehydrogenase is loosely bound to the inner mitochondrial membrane. The NADH:ubiquinone reductase activity is insensitive to rotenone, flavone, or dicumarol; however, it was inhibited by diphenyl iodonium in a time- and concentration-dependent manner. Maximum inhibition by diphenyl iodonium required preincubation with NADH to reduce the flavin. More complete inhibition was obtained with the more hydrophobic electron acceptors, such as Q(1) or Q(2), as compared to the more hydrophilic ones, such as Q(0) or dichloroindophenol. Kinetic analysis of the enzyme indicated that the enzyme followed a ping-pong mechanism. The enzyme conducts a one-electron transfer and can reduce molecular oxygen forming superoxide radical. PMID- 11863446 TI - Topological stability and self-association of a completely hydrophobic model transmembrane helix in lipid bilayers. AB - Investigation of interactions between hydrophobic model peptides and lipid bilayers is perhaps the only way to elucidate the principles of the folding and stability of membrane proteins (White, S. H., and Wimley, W. C. (1998) Biochim. Biophys. Acta 1367, 339-352). We designed the completely hydrophobic "inert" peptide modeling a transmembrane (TM) helix without any of the specific side chain interactions expected, X-(LALAAAA)(3)-NH(2) [X = Ac (I), 7-nitro-2-1,3 benzoxadiazol-4-yl (II), or 5(6)-carboxytetramethylrhodamine (III)]. Fourier transform infrared-polarized attenuated total reflection measurements revealed that I as well as II assume a TM helix in hydrated 1-palmitoyl-2-oleoyl-sn glycero-3-phosphocholine bilayers. Dithionite quenching experiments detected no topological change (flip-flop) in the helix II for at least 24 h. Thus, the TM helix itself is a highly stable structure, even in the absence of flanking hydrophilic or aromatic amino acids which are suggested to play important roles in stable TM positioning. Helix self-association in lipid bilayers was detected by fluorescence resonance energy transfer between II and III. The peptide was in a monomer-antiparallel dimer equilibrium with an association free energy of approximately -13 kJ/mol. Electron spin resonance spectra of 1-palmitoyl-2 stearoyl-(14-doxyl)-sn-glycero-3-phosphocholine demonstrated the presence of a motionally restricted component at lower temperatures. PMID- 11863447 TI - High yield of B-branch electron transfer in a quadruple reaction center mutant of the photosynthetic bacterium Rhodobacter sphaeroides. AB - A new reaction center (RC) quadruple mutant, called LDHW, of Rhodobacter sphaeroides is described. This mutant was constructed to obtain a high yield of B branch electron transfer and to study P(+)Q(B)(-) formation via the B-branch. The A-branch of the mutant RC contains two monomer bacteriochlorophylls, B(A) and beta, as a result of the H mutation L(M214)H. The latter bacteriochlorophyll replaces bacteriopheophytin H(A) of wild-type RCs. As a result of the W mutation A(M260)W, the A-branch does not contain the ubiquinone Q(A); this facilitates the study of P(+)Q(B)(-) formation. Furthermore, the D mutation G(M203)D introduces an aspartic acid residue near B(A). Together these mutations impede electron transfer through the A-branch. The B-branch contains two bacteriopheophytins, Phi(B) and H(B), and a ubiquinone, Q(B.) Phi(B) replaces the monomer bacteriochlorophyll B(B) as a result of the L mutation H(M182)L. In the LDHW mutant we find 35-45% B-branch electron transfer, the highest yield reported so far. Transient absorption spectroscopy at 10 K, where the absorption bands due to the Q(X) transitions of Phi(B) and H(B) are well resolved, shows simultaneous bleachings of both absorption bands. Although photoreduction of the bacteriopheophytins occurs with a high yield, no significant (approximately 1%) P(+)Q(B)(-) formation was found. PMID- 11863448 TI - Molecular alignment of denatured states of staphylococcal nuclease with strained polyacrylamide gels and surfactant liquid crystalline phases. AB - Residual dipolar couplings reflect the orientation of vectors between pairs of magnetic nuclei relative to a unique set of molecular axes. Thus, unlike NOEs and scalar couplings, dipolar couplings provide access to long-range structural information. A prerequisite for measurement of these NMR parameters is imposition of a weak net alignment, most simply by forcing the macromolecules to tumble in an asymmetric environment that restricts some orientations more than others. In this report, several denatured forms of staphylococcal nuclease are aligned by using compressed and stretched polyacrylamide gels, a nonionic type of lipid bilayer disk or bicelle, and a liquid crystalline phase formed by a cationic lipid. All three types of media can be used at high urea concentrations. While polyacrylamide gels and bicelles produce similar alignment tensors through steric interactions, a liquid crystalline phase of cetylpyridinium bromide aligns denatured nuclease along a different set of axes, presumably through electrostatic effects. The analysis of residual dipolar couplings collected with two different alignment tensors may permit the calculation of ensembles of conformations. The dipolar couplings observed for staphylococcal nuclease denatured with urea, by low pH or by deletion of residues from both termini, suggest that all denatured forms share a common "topology", one which has been shown previously to be native-like. Although SDS/nuclease complexes give sharp and disperse (1)H-(15)N correlation spectra, only small couplings are observed in strained polyacrylamide gels. PMID- 11863449 TI - A hyperthermophilic plant-type [2Fe-2S] ferredoxin from Aquifex aeolicus is stabilized by a disulfide bond. AB - A [2Fe-2S] ferredoxin (Fd1) from the hyperthermophilic bacterium Aquifex aeolicus has been obtained by heterologous expression of the encoding gene in Escherichia coli. Sequence comparisons show that this protein belongs to the extended family of plant- and mammalian-type [2Fe-2S] ferredoxins but also indicate that it is not closely similar to either the plant-type or mammalian-type subfamilies. Instead, it appears to bear some similarity to novel members of this family, in particular the Isc-type ferredoxins involved in the assembly of iron-sulfur clusters in vivo. The two redox levels of the [2Fe-2S](2+/+) metal site of A. aeolicus ferredoxin have been studied by UV-visible, resonance Raman, EPR, variable temperature magnetic circular dichroism, and Mossbauer spectroscopies. A full-spin Hamiltonian analysis is given for the Mossbauer spectra. In aggregate, the spectroscopic data reveal differences with both the plant-type and mammalian type ferredoxins, in keeping with the sequence comparisons. The midpoint potential of the [2Fe-2S](2+/+) couple, at -375 mV versus the normal hydrogen electrode, is more negative than those of mammalian-type ferredoxins and at the upper end of the range covered by plant-type ferredoxins. A. aeolicus ferredoxin contains two cysteines in addition to the four that are committed as ligands of the [2Fe-2S] cluster. These two residues have been shown by chemical modification and site-directed mutagenesis to form a disulfide bridge in the native protein. While that cystine unit plays a significant role in the exceptional thermostability of A. aeolicus ferredoxin (T(m) = 121 degrees C at pH 7 versus T(m) = 113 degrees C in a molecular variant where the disulfide bridge has been removed), it does not bear on the properties of the [2Fe-2S](2+/+) chromophore. This observation is consistent with the large distance (ca. 20 A) that is predicted to separate the iron-sulfur chromophore from the disulfide bridge. PMID- 11863450 TI - Intrastrand DNA cross-links as tools for studying DNA replication and repair: two , three-, and four-carbon tethers between the N(2) positions of adjacent guanines. AB - A general protocol for preparation of oligonucleotides containing intrastrand cross-links between the exocyclic amino groups of adjacent deoxyguanosines has been developed. A series of 2, 3, and 4 methylene cross-links was incorporated site-specifically into an 11-mer (5'-GGCAGGTGGTG-3', cross-linked positions are underlined) via a reaction between oligonucleotide containing 2-fluoro-O(6) trimethylsilylethyl deoxyinosines and the appropriate diamine (ethylenediamine, 1,3-diaminopropane, 1,4-diaminobutane). These cross-linked-oligonucleotides were studied for their ability to bend DNA by the method of Koo and Crothers [Koo, H. S., and Crothers, D. M. (1988) Proc. Natl. Acad. Sci. U.S.A. 85, 1763-1767] in which the mobility of ligated oligomers in nondenaturing polyacrylamide gels is evaluated. It was found that all cross-links induced bending (2-carbon cross link, 30.0 +/- 4.0 deg/turn; 3-carbon cross-link, 11.7 +/- 1.6 deg/turn; 4-carbon cross-link, 7.4 +/- 1.0 deg/turn). Despite the differing extent of helical distortion exhibited by the cross-links, all appeared to be equally blocking to replication by the Escherichia coli polymerases, pol I, pol II, and pol III. In contrast, when incision of the cross-links by the E. coli UvrABC nucleotide incision complex was studied, the extent of incision of the cross-link was found to correlate closely with the degree of bending measured in the gel mobility assay, i.e., the efficiency of incision was 2-carbon >> 3-carbon > 4-carbon. PMID- 11863451 TI - Omega-3 triglycerides modify blood clearance and tissue targeting pathways of lipid emulsions. AB - Omega-3-rich (n-3) triglycerides (TG) are increasingly recognized as having modulating roles in many physiological and pathological conditions. We questioned whether the catabolism of lipid emulsions would be changed after enrichment with fish oil (n-3) TG as compared to enrichment with omega-6-rich soy oil (n-6) TG. Phospholipid-stabilized emulsions of n-3 TG and n-6 TG were labeled with [(3)H]cholesteryl oleoyl ether and administered by bolus injection to wild-type (WT) mice, mice lacking the low-density lipoprotein receptor (LDL-R) (LDL-R -/-), and apolipoprotein E (apoE) knockout mice (apoE -/-). The effects of exogenous apoE, heparin, Triton WR 1339, and lactoferrin on catabolism of emulsions were also assayed. n-3 TG emulsions were cleared faster from blood and had different extrahepatic tissue targeting compared to n-6 TG emulsions. In apoE -/- and LDL-R -/- mice, blood clearance of n-6 TG emulsions slowed with decreased liver uptake, but no changes were observed in n-3 TG emulsion clearance and tissue uptake compared to WT mice. In WT mice, addition of exogenous apoE to the emulsion increased liver uptake of n-6 TG emulsions but had no impact on n-3 TG emulsions. Pre-injection of heparin increased and Triton WR 1339 and lactoferrin decreased blood clearance of n-6 TG emulsions with little or no effect on n-3 TG emulsions. Liver uptake of n-6 TG emulsions increased after heparin injection and decreased after Triton WR 1339 injection, but uptake of n-3 TG emulsions was not changed. These data show that the catabolism of n-3 TG emulsions and the catabolism of n-6 TG emulsions occur via very different mechanisms. Removal of chylomicron-sized n 6 TG emulsions is modulated by lipoprotein lipase (LPL), apoE, LDL-R, and lactoferrin-sensitive pathways. In contrast, clearance of chylomicron-sized n-3 TG emulsions relies on LPL to a very minor extent and is independent of apoE, LDL R, and lactoferrin-sensitive pathways. PMID- 11863452 TI - Glu11 site cleavage and N-terminally truncated A beta production upon BACE overexpression. AB - Amyloid beta peptides (A beta) are generated by the proteolytic processing of the amyloid beta precursor protein (APP). The newly identified beta-site APP-cleaving enzyme (BACE) cleaves APP at Asp1 as well as between Tyr10 and Glu11 of A beta, producing C-terminal fragments (CTFs) C99 and C89, respectively. Subsequent cleavage by gamma-secretase gives rise to A beta 1-40/42 and A beta 11-40/42. Although both full-length and A beta peptides truncated at residue 11 have been identified in amyloid plaques in the AD brain, the relative proportion of these two cleavage products produced by BACE and secreted into the medium by cultured cells is unknown. Using cell lines stably overexpressing BACE, we found that A beta 11-40 and A beta 11-42 are major A beta cleavage products generated by BACE. We further showed that BACE utilizes both full-length APP as well as C99 as substrates for the production of C89, and that A beta 11-40/42 can be generated by sequential cleavage of single APP molecules by BACE and gamma-secretase. Taken together, the abundance of A beta 11-40/42 produced by BACE suggests that their roles in AD pathogenesis may be underestimated. PMID- 11863453 TI - Limited topological alteration of the T7 RNA polymerase active center at intrinsic termination sites. AB - Transcription terminators trigger the dissociation of RNA polymerase elongation complexes and the release of RNA products at specific DNA template positions. The mechanism by which these signals alter the catalytic properties of the highly processive elongation transcription complexes is unclear. Here, we propose that intrinsic terminators impede transcript elongation by promoting a misarrangement of reactants and catalytic effectors within the active site of T7 RNA polymerase. In effect, a productive catalytic coordination network can be readily restored when Mg(2+) effectors are replaced by the more "relaxing" Mn(2+) ions, leading to transcript elongation beyond the termination point. This Mn(2+)-dependent incorporation of additional nucleotides occurs within unstable transcription complexes that ultimately dissociate at positions downstream from the normal termination site. Thus, Mn(2+) coordination in the polymerase active center can compensate for the disruptive but limited perturbation of the catalytic arrangement of reactants that accompany larger structural changes of the transcription complex triggered by termination signals. These results provide evidence that the geometry of the catalytic coordination network within the active site is a crucial component of RNA polymerase catalysis. Limited variations of the active site architecture are sufficient to confer tight control of the RNA polymerase function and, thus, may ubiquitously benefit signals regulating transcription. PMID- 11863454 TI - Gal4-VP16 and Gal4-AH increase the orientational and axial specificity of TATA box recognition by TATA box binding protein. AB - Previous work has shown that binding of the TATA box binding protein (TBP) to the TATA box is a rate-limiting step during pre-initiation complex (PIC) formation. Although the transcription of eukaryotic genes normally proceeds in one direction, studies in solution have shown that TBP lacks the information necessary to orient itself on the TATA box. Instead, yeast TBP binds TATA containing promoters in two orientations that are related by a 180 rotation about TBP's pseudo-2-fold symmetry axis. Recruitment of PIC components by gene-specific activators is considered a primary mechanism of transcriptional enhancement. Here we ask whether activators might function, at least in part, by increasing the fraction of PICs assembled with TBP bound in the orientation necessary for transcription. We use DNA affinity cleavage and a TBP-phenanthroline-copper conjugate to monitor the orientation of TBP in the presence of the well-studied activators Gal4-VP16 and Gal4-AH. In the absence of a transcriptional activator, only 51% of the TBP x TATA box complexes were bound in the orientation necessary for the initiation of transcription. However, in the presence of saturating Gal4 VP16, 87% of the TBP bound to the TATA box was oriented correctly at equilibrium. This increase in orientational specificity corresponds to a free energy difference (Delta Delta G(obs)) of 1.1 kcal x mol(-1) and was accompanied by a dramatic increase in axial specificity, reminiscent of the effects of transcription factors TFIIB and TFIIA reported previously. Gal4-AH also enhanced the orientational and axial specificity of the TBP x TATA complex, although to a lesser extent. We suggest that these effects on specificity represent a variation of recruitment, since they require direct interactions between the activator and a PIC component but only increase the effective concentration of the correctly oriented PIC component. These findings add to increasing evidence that recruitment may encompass a broad range of mechanisms. PMID- 11863455 TI - Carcinogenic chromium(VI) induces cross-linking of vitamin C to DNA in vitro and in human lung A549 cells. AB - Reductive activation of carcinogenic Cr(VI) is required for the induction of DNA damage and mutations. Here, we examined the formation of Cr-DNA adducts in the reactions of Cr(VI) with its dominant biological reducer, vitamin C (ascorbate). Reductive conversion of Cr(VI) to Cr(III) by ascorbate produced stable Cr-DNA adducts, of which approximately 25% constituted ascorbate-Cr(III)-DNA cross links. No evidence was found for the involvement of Cr(V) or Cr(IV) intermediates in the formation of either binary or ternary adducts. The cross-linking reaction was consistent with the attack of DNA by transient Cr(III)-ascorbate complexes. The yield of Cr(III)-DNA adducts was similar on dsDNA and AGT, ACT, or CT oligonucleotides and was strongly inhibited by Mg(2+), suggesting predominant coordination of Cr(III) to DNA phosphate oxygens. We also detected cross-linking of ascorbate to DNA in Cr(VI)-exposed human lung A549 cells that were preincubated with dehydroascorbic acid to create normal levels of intracellular ascorbate. Ascorbate-Cr-DNA cross-links accounted for approximately 6% of the total Cr-DNA adducts in A549 cells. Shuttle-vector experiments showed that ascorbate-Cr-DNA cross-links were mutagenic in human cells. Our results demonstrate that in addition to reduction of Cr(VI) to DNA-reactive Cr(III), vitamin C contributes to the genotoxicity of Cr(VI) via a direct chemical modification of DNA. The absence of Asc in A549 and other human cultured cells indicates that cells maintained under the usual in vitro conditions lack the most important reducing agent for Cr(VI) and would primarily display slow thiol dependent activation of Cr(VI). PMID- 11863456 TI - Structural requirements for high-affinity heparin binding: alanine scanning analysis of charged residues in the C-terminal domain of human extracellular superoxide dismutase. AB - An essential property of human extracellular superoxide dismutase (hEC-SOD) is its affinity for heparin and heparan sulfate proteoglycans located on cell surfaces and in the connective tissue matrix. The C-terminal domain of hEC-SOD plays the major role in this interaction. This domain has an unusually high content of charged amino acids: six arginine, three lysine, and five glutamic acid residues. In this study, we used alanine scanning mutagenesis of charged amino acids in the C-terminal domain to elucidate the requirements for the heparin/heparan sulfate interaction. As a tool in this study, we used a fusion protein comprising the C-terminal domain of hEC-SOD fused to human carbonic anhydrase II (HCAII). The interaction studies were performed using the surface plasmon resonance technique and heparin-Sepharose chromatography. Replacement of the glutamic acid residues by alanine resulted, in all cases, in tighter binding. All alanine substitutions of basic amino acid residues, except one (R205A), reduced heparin affinity. The arginine and lysine residues in the cluster of basic amino acid residues (residues 210-215), the RK-cluster, are of critical importance for the binding to heparin, and arginine residues promote stronger interactions than lysine residues. PMID- 11863457 TI - Combined participation of hydroxylase active site residues and effector protein binding in a para to ortho modulation of toluene 4-monooxygenase regiospecificity. AB - Toluene 4-monooxygenase (T4MO) is a diiron hydroxylase that exhibits high regiospecificity for para hydroxylation. This fidelity provides the basis for an assessment of the interplay between active site residues and protein complex formation in producing an essential biological outcome. The function of the T4MO catalytic complex (hydroxylase, T4moH, and effector protein T4moD) is evaluated with respect to effector protein concentration, the presence of T4MO electron transfer components (Rieske ferredoxin, T4moC, and NADH oxidoreductase), and use of mutated T4moH isoforms with different hydroxylation regiospecificities. Steady state kinetic analyses indicate that T4moC and T4moD form complexes of similar affinity with T4moH. At low T4moD concentrations, the steady-state hydroxylation rate is linearly dependent on T4moD-T4moH complex formation, whereas regiospecificity and the coupling efficiency between NADH consumption and hydroxylation are associated with intrinsic properties of the T4moD-T4moH complex. The optimized complex gives both efficient coupling and high regiospecificity with p-cresol representing >96% of total products from toluene. Similar coupling and regiospecificity for para hydroxylation are obtained with T3buV (an effector protein from a toluene 3-monooxygenase), demonstrating that effector protein binding does not uniquely determine or alter the regiospecificity of toluene hydroxylation. The omission of T4moD causes an approximately 20-fold decrease in hydroxylation rate, nearly complete uncoupling, and a decrease in regiospecificity so that p-cresol represents approximately 60% of total products. Similar shifts in regiospecificity are observed in oxidations of alternative substrates in the absence or upon the partial removal of either T4moD or T3buV from toluene oxidations. The mutated T4moH isoforms studied have apparent V(max)/K(M) specificities differing by approximately 2-4-fold and coupling efficiencies ranging from 88% to 95%, indicating comparable catalytic function, but also exhibit unique regiospecificity patterns for all substrates tested, suggesting unique substrate binding preferences within the active site. The G103L isoform has enhanced selectivity for ortho hydroxylation with all substrates tested except nitrobenzene, which gives only m-nitrophenol. The regiospecificity of the G103L isoform is comparable to that observed from naturally occurring variants of the toluene/benzene/o-xylene monooxygenase subfamily. Evolutionary and mechanistic implications of these findings are considered. PMID- 11863458 TI - The serpin SQN-5 is a dual mechanistic-class inhibitor of serine and cysteine proteinases. AB - SQN-5 is a mouse serpin that is highly similar to the human serpins SCCA1 (SERPINB3) and SCCA2 (SERPINB4). Previous studies characterizing the biochemical activity of SQN-5 showed that this serpin, like SCCA2, inhibited the chymotrypsin like enzymes mast cell chymase and cathepsin G. Using an expanded panel of papain like cysteine proteinases, we now show that SQN-5, like SCCA1, inhibited cathepsins K, L, S, and V but not cathepsin B or H. These interactions were characterized by stoichiometries of inhibition that were nearly 1:1 and second order rate constants of >10(4) M(-1) s(-1). Reactive site loop (RSL) cleavage analysis showed that SQN-5 employed different reactive centers to neutralize the serine and cysteine proteinases. To our knowledge, this is the first serpin that serves as a dual inhibitor of both chymotrypsin-like serine and the papain-like cysteine proteinases by employing an RSL-dependent inhibitory mechanism. The ability of serpins to inhibit both serine and/or papain-like cysteine proteinases may not be a recent event in mammalian evolution. Phylogenetic studies suggested that the SCCA and SQN genes evolved from a common ancestor approximately 250-280 million years ago. When the fact that mammals and birds diverged approximately 310 million years ago is considered, an ancestral SCCA/SQN-like serpin with dual inhibitory activity may be present in many mammalian genomes. PMID- 11863459 TI - A novel reaction between adenosylcobalamin and 2-methyleneglutarate catalyzed by glutamate mutase. AB - We describe a novel reaction of adenosylcobalamin that occurs when adenosylcobalamin-dependent glutamate mutase is reacted with the substrate analogue 2-methyleneglutarate. Although 2-methyleneglutarate is a substrate for the closely related adenosylcobalamin-dependent enzyme 2-methyleneglutarate mutase, it reacts with glutamate mutase to cause time-dependent inhibition of the enzyme. Binding of 2-methyleneglutarate to glutamate mutase initiates homolysis of adenosylcobalamin. However, instead of the adenosyl radical proceeding to abstract a hydrogen from the substrate, which is the next step in all adenosylcobalamin-dependent enzymes, the adenosyl radical undergoes addition to the exo-methylene group to generate a tertiary radical at C-2 of methyleneglutarate. This radical has been characterized by EPR spectroscopy with regiospecifically (13)C-labeled methyleneglutarates. Irreversible inhibition of the enzyme appears to be a complicated process, and the detailed chemical and kinetic mechanism remains to be elucidated. The kinetics of this process suggest that cob(II)alamin may reduce the enzyme-bound organic radical so that stable adducts between the adenosyl moiety of the coenzyme and 2-methyleneglutarate are formed. PMID- 11863460 TI - Alkaline phosphatase revisited: hydrolysis of alkyl phosphates. AB - Escherichia coli alkaline phosphatase (AP) is the prototypical two metal ion catalyst with two divalent zinc ions bound approximately 4 A apart in the active site. Studies spanning half a century have elucidated many structural and mechanistic features of this enzyme, rendering it an attractive model for investigating the potent catalytic power of bimetallic centers. Unfortunately, fundamental mechanistic features have been obscured by limitations with the standard assays. These assays generate concentrations of inorganic phosphate (P(i)) in excess of its inhibition constant (K(i) approximately 1 muM). This tight binding by P(i) has affected the majority of published kinetic constants. Furthermore, binding limits k(cat)/K(m) for reaction of p-nitrophenyl phosphate, the most commonly employed substrate. We describe a sensitive (32)P-based assay for hydrolysis of alkyl phosphates that avoids the complication of product inhibition. We have revisited basic mechanistic features of AP with these alkyl phosphate substrates. The results suggest that the chemical step for phosphorylation of the enzyme limits k(cat)/K(m). The pH-rate profile and additional results suggest that the serine nucleophile is active in its anionic form and has a pK(a) of < or = 5.5 in the free enzyme. An inactivating pK(a) of 8.0 is observed for binding of both substrates and inhibitors, and we suggest that this corresponds to ionization of a zinc-coordinated water molecule. Counter to previous suggestions, inorganic phosphate dianion appears to bind to the highly charged AP active site at least as strongly as the trianion. The dependence of k(cat)/K(m) on the pK(a) of the leaving group follows a Bronsted correlation with a slope of beta(lg) = -0.85 +/- 0.1, differing substantially from the previously reported value of -0.2 obtained from data with a less sensitive assay. This steep leaving group dependence is consistent with a largely dissociative transition state for AP-catalyzed hydrolysis of phosphate monoesters. The new (32)P-based assay employed herein will facilitate continued dissection of the AP reaction by providing a means to readily follow the chemical step for phosphorylation of the enzyme. PMID- 11863461 TI - The (alpha F(357)C)(3)(beta R(372)C)(3)gamma subcomplex of the F(1)-ATPase from the thermophilic Bacillus PS3 has altered ATPase activity after cross-linking alpha and beta subunits at noncatalytic site interfaces. AB - In crystal structures of bovine MF(1), the side chains of alpha F(357) and beta R(372) are near the adenines of nucleotides bound to noncatalytic sites. To determine if during catalysis these side chains must pass through the different arrangements in which they are present in crystal structures, the catalytic properties of the (alpha F(357)C)(3)(beta R(372)C)(3)gamma subcomplex of the TF(1)-ATPase were characterized before and after cross-linking the introduced cysteines with CuCl(2). The unmodified mutant enzyme hydrolyzes MgATP at 50% the rate exhibited by wild type. Detailed comparison of the catalytic properties of the double mutant enzyme before and after cross-linking with those of the wild type subcomplex revealed the following. Before cross-linking, the (alpha F(357)C)(3)(beta R(372)C)(3)gamma subcomplex has less tendency than wild type to release inhibitory MgADP entrapped in a catalytic site during turnover when MgATP binds to noncatalytic sites. Following cross-linking, ATPase activity is reduced 5-fold, and inhibitory MgADP entrapped in a catalytic site during turnover does not release under conditions wherein binding of ATP to noncatalytic sites of the wild-type enzyme promotes release of MgADP from the affected catalytic site. When assayed in the presence of lauryldimethylamine oxide, which prevents turnover dependent entrapment of inhibitory MgADP in a catalytic site, ATPase activity of the cross-linked form is 47% that of the unmodified mutant enzyme. These results suggest that, during catalysis, the side chains of alpha F(357) and beta R(372) do not pass through the extremely different relative positions in which they exist at the three noncatalytic site interfaces in crystal structures. PMID- 11863462 TI - Chemical rescue in catalysis by human carbonic anhydrases II and III. AB - The maximal velocity of catalysis of CO(2) hydration by human carbonic anhydrase II (HCA II) requires proton transfer from zinc-bound water to solution assisted by His 64. The catalytic activity of a site-specific mutant of HCA II in which His 64 is replaced with Ala (H64A HCA II) can be rescued by exogenous proton donors/acceptors, usually derivatives of imidazole and pyridine. X-ray crystallography has identified Trp 5 as a binding site of the rescue agent 4 methylimidazole (4-MI) on H64A HCA II. This binding site overlaps with the "out" position in which His 64 in wild-type HCA II points away from the zinc. Activation by 4-MI as proton donor/acceptor in catalysis was determined in the dehydration direction using (18)O exchange between CO(2) and water and in the hydration direction by stopped-flow spectrophotometry. Replacement of Trp 5 by Ala, Leu, or Phe in H64A HCA II had no significant effect on enhancement by 4-MI of maximal rate constants for proton transfer in catalysis to levels near 10(5) s(-1). This high activity for chemical rescue indicates that the binding site of 4-MI at Trp 5 in H64A HCA II appears to be a nonproductive binding site, although it is possible that a similarly effective pathway for proton transfer exists in the mutants lacking Trp 5. Moreover, the data suggest that the out position of His 64 considered alone is not active in proton transfer in HCA II. In contrast to isozyme II, the replacement of Trp 5 by Ala in HCA III abolished chemical rescue of k(cat) by imidazole but left k(cat)/K(m) for hydration unchanged. This demonstrates that Trp 5 contributes to the predominant productive binding site for imidazole, with a maximal level for the rate constant of proton transfer near 10(4) s(-1). This difference in the susceptibility of CA II and III to chemical rescue may be related to the more sterically constrained and electrostatically positive nature of the active site cavity of CA III compared with CA II. The possibility of nonproductive binding sites for exogenous proton donors offers an explanation for the unusually low value of the intrinsic kinetic barrier obtained by application of Marcus theory to chemical rescue of H64A HCA II. PMID- 11863463 TI - Interaction of the membrane-inserted diphtheria toxin T domain with peptides and its possible implications for chaperone-like T domain behavior. AB - The T domain of diphtheria toxin is believed to aid the low-pH-triggered translocation of the partly unfolded A chain (C domain) through cell membranes. Recent experiments have suggested the possibility that the T domain aids translocation by acting as a membrane-inserted chaperone [Ren, J., et al. (1999) Science 284, 955-957]. One prediction of this model is that the membrane-inserted T domain should be able to interact with sequences that mimic unfolded proteins. To understand the basis of interaction of the membrane-inserted T domain with unfolded polypeptides, its interaction with water-soluble peptides having different sequences was studied. The membrane-inserted T domain was able to recognize helix-forming 23-residue Ala-rich peptides. In the presence of such peptides, hydrophobic helix 9 of the T domain underwent the previously characterized conformational change from a state exhibiting shallow membrane insertion to one exhibiting deep insertion. This conformational change was more readily induced by the more hydrophobic peptides that were tested. It did not occur at all in the presence a hydrophilic peptide in which alternating Ser and Gly replaced Ala or in the presence of unfolded hydrophilic peptides derived from the A chain of the toxin. Interestingly, a peptide with a complex sequence (RKE(3)KE(2)LMEW(2)KM(2)SETLNF) also interacted with the T domain very strongly. We conclude that the membrane-inserted T domain cannot recognize every unfolded amino acid sequence. However, it does not exhibit strong sequence specificity, instead having the ability to recognize and interact with a variety of amino acid sequences having moderate hydrophobicity. This recognition was not strictly correlated with the strength of peptide binding to the lipid, suggesting that more than just hydrophobicity is involved. Although it does not prove that the T domain functions as a chaperone, T domain recognition of hydrophobic sequences is consistent with it having polypeptide recognition properties that are chaperone like. PMID- 11863464 TI - Single replacement constructs of all hydroxyl, basic, and acidic amino acids identify new function and structure-sensitive regions of the mitochondrial phosphate transport protein. AB - The phosphate transport protein (PTP) catalyzes the proton cotransport of phosphate into the mitochondrial matrix. It functions as a homodimer, and thus residues of the phosphate and proton pores are somewhat scattered throughout the primary sequence. With 71 new single mutation per subunit PTPs, all its hydroxyl, basic, and acidic residues have now been replaced to identify these essential residues. We assayed the initial rate of pH gradient-dependent unidirectional phosphate transport activity and the liposome incorporation efficiency (LIE) of these mutants. Single mutations of Thr79, Tyr83, Lys90, Tyr94, and Lys98 inactivate transport. The spacings between these residues imply that they are located along the same face of transmembrane (TM) helix B, requiring an extension of its current model C-terminal domain by 10 residues. This extension superimposes very well onto the shorter bovine PTP helix B, leaving a 15-residue hydrophobic extension of the yeast helix B N-terminus. This is similar to the helix D and F regions of the yeast PTP. Only one transport-inhibiting mutation is located within loops: Ser158Thr in the matrix loop between helices C and D. All other transport-inhibiting mutations are located within the TM helices. Mutations that yield LIEs of <6% are all, except for four, within helices. The four exceptions are Tyr12Ala near the PTP N-terminus and Arg159Ala, Glu163Gln, and Glu164Gln in the loop between helices C and D. The PTP C-terminal segment beyond Thr214 at the N-terminus of helix E has 11 mutations with LIEs >20% and none with LIE <6%. Mutations with LIEs >20% are located near the ends of all the TM helices except TM helix D. Only a few mutations alter PTP structure (LIE) and also affect PTP transport activity. A novel observation is that Ser4Ala blocks the formation of PTP bacterial inclusion bodies. PMID- 11863465 TI - Converting a maltose receptor into a nascent binuclear copper oxygenase by computational design. AB - Computational protein design methods were used to identify mutations that are predicted to introduce a binuclear copper center coordinated by six histidines, replacing the maltose-binding site in Escherichia coli maltose-binding protein (MBP) with an oxygen-binding site. A small family of five candidate designs consisting of 9 to 10 mutations each was constructed by oligonucleotide-directed mutagenesis. These mutant proteins were expressed and purified, and their stability, copper- and cobalt-binding properties, and interactions of the resulting metalloprotein complexes with azide, hydrogen peroxide, and dioxygen were characterized. We identified one 10-fold mutant, MBP.Hc.E, that can form Cu(II)(2) and Co(II)(2) complexes that interact with H(2)O(2) and O(2). The Co(II)(2) protein reacts with H(2)O(2) to form a complex that is spectroscopically similar to a synthetic model that structurally mimics the oxy hemocyanin core, whereas the Cu(II)(2) protein reacted with O(2) or H(2)O(2) does not. We postulate that the equilibrium between the open and closed conformations of MBP allows species with variable Cu-Cu distances to form, and that such species can bind ligands in geometries that are not observed in natural type III centers. Introduction of one additional mutation in the hinge region of MBP, I329F, known to favor formation of the closed state, results in a binuclear copper center that when reacted with low concentrations of H(2)O(2) mimics the spectroscopic signature of oxy-hemocyanin. PMID- 11863466 TI - Fibronectin peptides derived from two distinct regions stimulate adipocyte differentiation by preventing fibronectin matrix assembly. AB - Here, we show that fibronectin (FN) peptides derived from two distinct regions promote the insulin-induced adipocyte differentiation of ST-13 cells by preventing FN fibrillogenesis. ST-13 cells formed numerous FN fibrils under nonadipogenic conditions, whereas this FN fibrillogenesis was suppressed by adipose induction with insulin. The insulin-induced adipocyte differentiation was promoted by an amino-terminal 24-kDa fragment of FN, accompanied by further suppression of FN fibrillogenesis. The 24 K fragment prevented FN matrix assembly by direct incorporation into the FN matrix. Like the 24 K fragment, a peptide from the 14th type III repeat, termed FNIII14, which suppressed the integrin alpha 5 beta 1-mediated adhesion of ST-13 cells to FN, accelerated the adipocyte differentiation by preventing FN fibrillogenesis without direct incorporation into the FN matrix. FNIII14 induced the conformation change of beta1 integrins of K562 cells from active to resting, as judged by FACS analysis using a monoclonal antibody AG89 directed to an active beta1 integrin-dependent epitope. Binding of a (125)I-labeled FN fragment containing the RGD cell adhesive site to ST-13 cell surface was dissociated by FNIII14, with a concomitant binding of FNIII14 itself to the cell surface. The affinity labeling of ST-13 cells using biotinylated FNIII14 showed that FNIII14 specifically bound to a nonintegrin membrane protein with M(r) of around 50 kDa. Thus, the results indicated that prevention of FN fibrillogenesis by the 24 K Fib 1 fragment and FNIII14 caused the promotion of adipocyte differentiation of ST-13 cells and that the former was due to the direct incorporation into the FN matrix and that the latter might be interpreted by negative regulation of FN receptor alpha 5 beta 1 activity. PMID- 11863468 TI - Superior physicians and the treatment of hypertension. PMID- 11863469 TI - What is C-reactive protein telling us about coronary artery disease? PMID- 11863470 TI - Integrative medicine: bringing medicine back to its roots. PMID- 11863471 TI - Sensitivity and specificity of ultrasonography in the diagnosis of upper extremity deep vein thrombosis: a systematic review. AB - OBJECTIVES: To determine the sensitivity and specificity of ultrasonography in the diagnosis of upper extremity deep vein thrombosis and to determine the safety of withholding anticoagulant therapy in patients with negative ultrasonographic results. DATA SOURCES: The MEDLINE database was searched for literature published from January 1, 1980, to December 31, 2000, that evaluated ultrasonography for the diagnosis of upper extremity deep vein thrombosis. Bibliographies of the retrieved articles were cross-checked to identify additional studies. STUDY SELECTION: All prospective English-language studies were selected. Retrospective studies, review articles, and case reports were excluded. DATA EXTRACTION: Two of us (B.O.M. and S.W.R.) used predefined criteria to independently assess each study. Data on sensitivity and specificity and the associated 95% confidence intervals were recorded when available. DATA SYNTHESIS: Only one study met all of the predefined criteria for adequately evaluating sensitivity and specificity. The sensitivity of duplex ultrasonography ranged from 56% to 100%, and the specificity ranged from 94% to 100%. No study evaluated the safety of withholding anticoagulant therapy without additional testing in patients with negative ultrasonographic results. CONCLUSION: The safety of withholding anticoagulant treatment in a patient with suspected upper extremity deep vein thrombosis and negative ultrasonographic results is uncertain. PMID- 11863473 TI - Physician-related barriers to the effective management of uncontrolled hypertension. AB - BACKGROUND: Primary care physicians may not be aggressive enough with the management of hypertension. The purpose of this study was to identify barriers to primary care physicians' willingness to increase the intensity of treatment among patients with uncontrolled hypertension. METHODS: Descriptive survey study. We sampled patient visits in a large midwestern health system to identify patients with uncontrolled hypertension. The treating primary care physicians were asked to complete a survey about the patient visit with a copy of the office notes attached to the survey (patient visits, n = 270; response rate, 86%). RESULTS: Pharmacologic therapy was initiated or changed at only 38% of visits, despite documented hypertension for at least 6 months before the patients' most recent visit. The most frequently cited reason for no initiation or change in therapy was related to the primary care physicians being satisfied with the blood pressure (BP) value (satisfactory BP response, 30%; satisfactory diastolic BP response, 16%; only borderline hypertension, 10%). At 93% of these visits, systolic BP values were 140 mm Hg or higher, which is above the cut point recommended by Sixth Report of the Joint National Committee on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure guidelines, and 35% were 150 mm Hg or higher. On average, physicians reported that 150 mm Hg was the lowest systolic BP at which they would recommend pharmacologic treatment to patients, compared with 91 mm Hg for diastolic BP. CONCLUSIONS: Our findings suggest that an important reason why physicians do not treat hypertension more aggressively is that they are willing to accept an elevated systolic BP in their patients. This has an important impact on public health because of the positive association between systolic BP and cardiovascular disease. PMID- 11863472 TI - Implications of cytochrome P450 interactions when prescribing medication for hypertension. AB - Many of the estimated 50 million Americans with high blood pressure receive medications for hypertension and for other conditions, placing them at risk for adverse drug interactions. The risk for hypertension and for adverse drug reactions is highest in the elderly, who have the greatest need for pharmacologic therapy. The most important class of drug interactions involves the cytochrome P450 microsomal enzyme system, which handles a variety of xenobiotic substances. A potential for interactions with these enzymes exists with calcium channel blockers, beta-adrenergic blocking agents, angiotensin-converting enzyme inhibitors, and angiotensin receptor blockers but not with diuretic antihypertensives, which are renally eliminated and more vulnerable to drug interactions that occur in the kidney. This article reviews the cytochrome P450 enzyme system, identifies drugs and foods that have been implicated in metabolic interactions with antihypertensive agents, and suggests measures for reducing the risk of adverse events when drugs are coadministered. PMID- 11863474 TI - Osteoporosis intervention following distal forearm fractures: a missed opportunity? AB - BACKGROUND: Fractures are a manifestation of osteoporosis, but therapeutic interventions to reduce the risk of recurrent fractures are not widespread. OBJECTIVE: To identify predictors of osteoporosis treatment in postmenopausal women following distal forearm fracture. METHODS: This population-based retrospective cohort study included all postmenopausal women, 45 years or older, residing in Olmsted County, Minnesota, who sustained a distal forearm fracture due to minimal trauma (a fall from standing height or under) in 1993 to 1997. Complete medical records were reviewed for each subject and Cox proportional hazards regression was used to evaluate the relationship of baseline demographic and clinical characteristics to therapeutic interventions for osteoporosis within 12 months following the fracture. RESULTS: A total of 343 women with a mean age of 70.5 years had a minimal trauma distal forearm fracture. Within 12 months, 83% had seen a nonorthopedic physician. Of these, 17% had a pharmacologic osteoporosis intervention and the 12-month actuarially estimated cumulative incidence of any intervention was 18% (95% confidence interval [CI], 14%-22%). In a multivariate analysis, treatment was more likely to be offered to those with a prior diagnosis of osteoporosis (relative risk [RR], 2.08; 95% CI, 1.21-3.58), previous distal forearm fracture (RR, 2.38; 95% CI, 1.30-4.34), or history of cigarette smoking (RR, 1.86; 95% CI, 1.11-3.12). CONCLUSIONS: Effective osteoporosis interventions are underutilized among postmenopausal women who experience an osteoporotic fracture. Further work is needed to overcome barriers to optimal osteoporosis management in these women who are at high risk for future complications of osteoporosis. PMID- 11863475 TI - Modifiable cardiovascular risk factors in adults with diabetes: prevalence and missed opportunities for physician counseling. AB - BACKGROUND: Cardiovascular disease (CVD) is the leading cause of death in adults with diabetes mellitus (DM). Counseling by physicians is effective in inducing lifestyle modification. OBJECTIVE: To determine the prevalence of modifiable CVD risk factors and counseling by physicians among adults with DM. METHODS: Data on 9496 adults with DM and 150 493 adults without DM from the 1999 Behavioral Risk Factor Surveillance System were analyzed to yield estimates of CVD risk factors and counseling by physicians during routine visits. Multiple logistic regression was used to adjust estimates for age, sex, ethnicity, education, and income. Population estimates were created using software for the statistical analysis of correlated data (SUDAAN) because of the complex survey design of the Behavioral Risk Factor Surveillance System. RESULTS: Diabetes mellitus was more prevalent in adults aged 55 and older and in blacks and Hispanic or other ethnicities (both P<.001). Modifiable CVD risk factors, such as hypertension (56% vs 22%), high cholesterol (41% vs 20%), obesity (78% vs 57%), and insufficient physical activity (66% vs 56%), were more prevalent in adults with DM (all P<.001) and differed by ethnicity, sex, and age. Counseling about weight loss (50% vs 21%, P<.001), smoking cessation (78% vs 67%, P =.01), eating less fat (78% vs 71%, P<.001), and increasing physical activity (67% vs 36%, P<.001) was less than ideal in both groups and did not change after adjusting for age, sex, ethnicity, education, and income with multiple logistic regression. CONCLUSIONS: Although adults with DM have a high prevalence of modifiable CVD risk factors, counseling by physicians about lifestyle modification is less than optimal. There is a need to improve patient counseling for lifestyle modification by primary care physicians. PMID- 11863476 TI - Evaluation of prescribing practices: risk of lactic acidosis with metformin therapy. AB - BACKGROUND: The risk of lactic acidosis during metformin therapy is linked to specific and well-documented conditions that constitute contraindications or precautions to use of the agent. We conducted a retrospective evaluation of metformin use to determine whether prescribing practices are in accord with published contraindications and precautions. METHODS: All patients admitted to the hospital during a 6-month period who received at least 1 dose of metformin were identified through hospital pharmacy records. Patient demographics and clinical characteristics were then evaluated to determine whether metformin was prescribed to patients possessing any of the risk factors associated with development of lactic acidosis. RESULTS: We identified 263 hospitalizations involving 204 patients who received at least 1 dose of metformin during inpatient admission. Patients had at least 1 absolute contraindication to metformin therapy in 71 admissions (27%). In 29 (41%) of these 71 admissions, treatment with metformin continued despite the contraindication. The most common contraindication, elevated serum creatinine concentration, was present or developed during 32 admissions (12%); however, metformin use was appropriately discontinued in only 8 (25%) of these 32 patients. Of the precautions against metformin use, concomitant administration of cationic agents was the most common, occurring in 97 admissions (37%). CONCLUSIONS: Many patients are treated with metformin despite having clinical conditions that place them at risk for developing lactic acidosis. To minimize this risk, it is essential that prescribers develop a better understanding of the prescribing guidelines for metformin. PMID- 11863477 TI - Association of tobacco smoking with goiter in a low-iodine-intake area. AB - BACKGROUND: Goiter development depends on genetic and environmental factors. The major environmental factor is iodine intake, whereas diverging results have been published concerning the association between smoking and goiter. METHODS: A comparable, cross-sectional study was performed of patients from 2 areas in Denmark with mild and moderate iodine deficiency. A random sample of women and men in selected age groups from the general community was investigated; 4649 subjects participated. Smoking habits were investigated with questionnaires and interviews. Ultrasonography and clinical examination of the thyroid were performed, serum thyroglobulin was measured, and iodine concentration in spot urine samples was analyzed. Data were analyzed in linear models and logistic regression analyses. RESULTS: Serum thyroglobulin level and thyroid volume at ultrasonography were positively associated with smoking habits (P<.001); the association was stronger in the area with the lowest iodine intake (interaction: P<.001 for thyroglobulin, P =.04 for thyroid volume). A positive association with smoking was also found for thyroid enlargement (odds ratio, 2.9; 95% confidence interval, 2.2-3.7) and palpable goiter (odds ratio, 3.1; 95% confidence interval, 1.6-5.8). Ex-smokers had a goiter prevalence close to that of never smokers. The fraction of goiter cases attributable to smoking was 49% (95% confidence interval, 29%-65%). CONCLUSIONS: Thyroid volume and goiter prevalence were closely associated with smoking habits, with the strongest association being found in the area with the most pronounced iodine deficiency. This may have implications for future goiter prevalences in Third World countries, with their increasing use of tobacco. Half of goiter cases in this population could be ascribed to smoking. PMID- 11863478 TI - Diagnosis of iron deficiency anemia in the elderly by transferrin receptor ferritin index. AB - BACKGROUND: The diagnosis of iron deficiency anemia (IDA) in the elderly is difficult because of the prevalence of chronic diseases, which can cause anemia with high ferritin levels, even in the presence of iron deficiency. Therefore, we studied the sensitivity and specificity of a serum transferrin receptor assay, which is not affected by chronic diseases, in the diagnosis of IDA in elderly patients. METHODS: We performed a prospective controlled study of 49 consecutive male and female patients older than 80 years who were admitted to an acute geriatric department. Bone marrow aspirate confirmed IDA in all 49 patients. Fourteen additional patients, also older than 80 years, with anemia but without evidence of iron deficiency on results of bone marrow examination, served as a control group. All patients underwent evaluation by means of a detailed medical history and results of complete physical examination, routine blood tests, and specific tests for diagnosis and evaluation of anemia. Examination of bone marrow aspirate was performed for all patients. Levels of transferrin receptor in serum were determined by means of a specific enzyme-linked immunosorbent assay. The transferrin receptor-ferritin index (TR-F index) was defined as the ratio of serum transferrin receptor level to log ferritin level. RESULTS: Only 8 patients could be diagnosed as having IDA by means of routine blood test results (serum iron, ferritin, and transferrin saturation levels). In contrast, the TR-F index disclosed IDA in 43 of the 49 patients, thus increasing the sensitivity from 16% to 88%. CONCLUSIONS: The diagnosis of IDA in the elderly by means of routine blood tests has a very low sensitivity. The TR-F index is much more sensitive, and when results are positive, the TR-F index can eliminate the need for bone marrow examination. PMID- 11863479 TI - Missed opportunities for prevention of osteoporotic fracture. AB - BACKGROUND: Osteoporotic fracture is a growing public health problem burden to society. Despite the importance of physician practices in preventing it, relatively little is known about the osteoporosis-related practices of US physicians. METHODS: A total of 1500 female members of a Connecticut independent practice association model health plan (aged 40-69 years) were surveyed to identify women's receipt of osteoporosis-related services (eg, prevention counseling, bone mineral density [BMD] testing, and communication about treatment options). These findings were compared with recommendations of the US Preventive Services Task Force and the National Osteoporosis Foundation. We received 1007 completed questionnaires, for a response rate of 69%. RESULTS: Only 49% of the sample reported that a health care provider ever discussed osteoporosis with them. In multivariate analyses, women with multiple risk factors were not more likely than other women to have been counseled about osteoporosis and its prevention, although those with an osteopenia/osteoporosis diagnosis were. In contrast to National Osteoporosis Foundation recommendations, only a small minority of high-risk women (12%-34%) had their BMD tested. Although most women with an osteopenia/osteoporosis diagnosis reported receiving information on estrogen replacement therapy, calcium, and weight-bearing exercise, fewer reported receiving information on pharmaceutical alternatives to estrogen (33%) and vitamin D (20%). CONCLUSIONS: The main trigger to physician counseling of women about osteoporosis and its prevention is an osteopenia/osteoporosis diagnosis. Women with multiple risk factors for osteoporosis are not being identified for preventive counseling interventions or BMD testing. PMID- 11863480 TI - Delirium predicts 12-month mortality. AB - BACKGROUND: Delirium has not been found to be a significant predictor of postdischarge mortality, but previous research has methodologic limitations including small sample sizes and inadequate control of confounding. This study aimed to determine the independent effects of presence of delirium, type of delirium (incident vs prevalent), and severity of delirium symptoms on 12-month mortality among older medical inpatients. METHODS: A prospective, observational study of 2 cohorts of medical inpatients was conducted with patients 65 years or older: 243 patients had prevalent or incident delirium, and 118 controls had no delirium. Baseline measures included presence of delirium and/or dementia, severity of delirium symptoms, physical function, comorbidity, and physiological and clinical severity of illness. Mortality during the 12 months after enrollment was analyzed with the Cox proportional hazards model with adjustment for covariates. RESULTS: The unadjusted hazard ratio of delirium with mortality was 3.44 (95% confidence interval, 2.05-5.75); the adjusted hazard ratio was 2.11 (95% confidence interval, 1.18-3.77). The effect of delirium was sustained over the entire 12-month period after adjustment for covariates and was stronger among patients without dementia. Among patients with dementia, there was a weak, nonsignificant effect of delirium on survival. After adjustment for covariates, mortality did not differ between patients with incident and prevalent delirium, but among patients with delirium without dementia, greater severity of delirium symptoms was associated with higher mortality. CONCLUSIONS: Delirium is an independent marker for increased mortality among older medical inpatients during the 12 months after hospital admission. It is a particularly important prognostic marker among patients without dementia. PMID- 11863481 TI - Prevalence of monoclonal gammopathy in patients with primary hyperparathyroidism: a prospective study. AB - BACKGROUND: The association between primary hyperparathyroidism (PHPT) and monoclonal gammopathy has been reported, but whether it is fortuitous remains unsettled. We conducted a prospective study to determine the prevalence of monoclonal gammopathies in patients with surgically proved PHPT. METHODS: In 101 consecutive patients with PHPT, serum immunoglobulins were systematically studied using agarose gel electrophoresis and immunofixation before and, when appropriate, after parathyroid surgery. The PHPT population was compared with a control series of patients with other diseases requiring surgery and with a group of patients with benign disease of the thyroid gland matched for age and sex to the PHPT population. RESULTS: Monoclonal immunoglobulin was detected in 10 (10%) of 101 patients with PHPT (including 2 with multiple myeloma) compared with 2 (2%) of 127 patients who underwent other surgery (P =.005) and 3 (3%) of 101 patients with benign thyroid diseases (P =.04). CONCLUSIONS: The prevalence of monoclonal gammopathies is high in patients with PHPT. At minimum, sensitive serum protein electrophoresis should be performed routinely in all patients with PHPT. Conversely, in patients with monoclonal gammopathy who have hypercalcemia but no other symptoms of progressive disease, clinicians must seek PHPT. PMID- 11863482 TI - Impact of a targeted intervention on lipid-lowering therapy in patients with coronary artery disease in the hospital setting. AB - BACKGROUND: Although lipid-lowering therapy according to the National Cholesterol Education Program guidelines decreases mortality and morbidity in patients with coronary artery disease (CAD), significant undertreatment of hyperlipidemia continues to occur. This study was designed to determine the impact of an intervention targeted at improving the use of lipid-lowering therapy in patients with CAD in the hospital setting. METHODS: Cardiac case managers prompted physicians to obtain lipid profiles for patients with CAD who were not receiving lipid-lowering therapy on admission and initiate lipid-lowering therapy for patients with a low-density lipoprotein level of 130 mg/dL (3.37 mmol/L) or higher during hospitalization. The study population comprised 813 patients with CAD admitted for percutaneous transluminal coronary angioplasty, coronary artery bypass grafting, or myocardial infarction. A retrospective chart review of lipid testing and treatment rates was conducted in 300 patients in the preintervention period, and a prospective review of rates was conducted in 513 patients during the intervention period. RESULTS: The percentage of patients with CAD not receiving lipid-lowering therapy on admission who had fractionated lipid profiles obtained during hospitalization increased from 27% preintervention to 89% during intervention (odds ratio, 18.27; 95% confidence interval, 11.61-28.74; P<.001). The percentage of patients with a low-density lipoprotein level of 130 mg/dL or higher for whom lipid-lowering therapy was initiated during hospitalization increased from 17% preintervention to 82% during intervention (odds ratio, 24.50; 95% confidence interval, 7.33-81.83; P<.001). CONCLUSIONS: The intervention provided by specialized cardiac case managers significantly increased physicians' adherence to the National Cholesterol Education Program treatment guidelines. The results of the present study suggest that intervention programs of this nature could produce a significant positive impact on cardiovascular outcomes if implemented nationally. PMID- 11863483 TI - Fulminant hepatitis and fatal toxic epidermal necrolysis (Lyell disease) coincident with clarithromycin administration in an alcoholic patient receiving disulfiram therapy. AB - Disulfiram is widely used in the treatment of chronic alcoholism. Adverse drug reactions with fatal outcome following disulfiram therapy are infrequent, and hepatic failure accounts for most of them. Since disulfiram is a cytochrome P450 (CYP450) enzyme system inhibitor, numerous interactions with several drugs metabolized in the liver have been reported. Like disulfiram, clarithromycin inhibits a CYP450 isoenzyme, but, despite its widespread use for the treatment of respiratory tract infections, no interactions with disulfiram have been described as yet. We report a case of fatal toxic epidermal necrolysis (Lyell disease) and fulminant hepatitis shortly after starting treatment with clarithromycin in a patient who was receiving disulfiram. This is the first case of such a severe dermatosis in a patient receiving either disulfiram or clarithromycin therapy. The temporal relationship between drug administration and clinical symptoms in this case suggests a probable interaction between the 2 drugs. PMID- 11863484 TI - Identification of Cardiobacterium hominis by broad-range bacterial polymerase chain reaction analysis in a case of culture-negative endocarditis. AB - Culture-negative bacterial endocarditis may be attributed to fastidious microorganisms, prior institution of antibiotic treatment, or both. We describe a case of culture-negative endocarditis in which a modified Steiner stain revealed bacterial structures in the resected heart valve material. Prompted by this finding, broad-range polymerase chain reaction (PCR) amplification of small subunit ribosomal DNA (16S rDNA) was performed, and Cardiobacterium hominis sequences were detected. This case demonstrates the usefulness of both the Steiner stain and broad-range direct molecular amplification as supplemental diagnostic tools in identification of otherwise unexplained infections. PMID- 11863485 TI - Aspirin, postmenopausal hormones, and C-reactive protein. PMID- 11863486 TI - Capillary leak syndrome after interferon treatment for chronic hepatitis C. PMID- 11863487 TI - Increased blood pressure variability may be associated with cognitive decline in hypertensive elderly subjects with no dementia. PMID- 11863489 TI - Metformin and vitamin B12 deficiency. PMID- 11863490 TI - New approaches to diagnosis and management of unstable angina and non-ST segment elevation myocardial infarction: controversial data. PMID- 11863492 TI - Driven lattice gas as a ratchet and pawl machine. AB - Boundary-induced transport in particle systems with anomalous diffusion exhibits rectification, negative resistance, and hysteresis phenomena depending on the way the drive acts on the boundary. The solvable case of a one-dimensional (1D) system characterized by a power-law diffusion coefficient and coupled to two particle reservoirs at different chemical potential is examined. In particular, it is shown that a microscopic realization of such a diffusion model is provided by a 3D driven lattice gas with kinetic constraints, in which energy barriers are absent and the local microscopic reversibility holds. PMID- 11863493 TI - Low autocorrelated multiphase sequences. AB - The interplay between the ground-state energy of the generalized Bernasconi model to multiphase, and the minimal value of the maximal autocorrelation function, C(max)=max(K)/C(K)/, K=1,...,N-1, is examined analytically in the thermodynamic limit where the main results are (a) For the binary case, the minimal value of C(max) over all sequences of length N, minC(max), is 0.435sqrt[N], significantly smaller than the typical value for random sequences O(sqrt[log N]sqrt[N]). (b) A new method to approximate F(max) is obtained using the observation of data collapse. (c) minC(max) is obtained in an energy which is about 30% above the ground-state energy of the generalized Bernasconi model, independent of the number of phases m. (d) For a given m, minC(max) proportional sqrt[N/m] indicating that for m=N, minC(max)=1, i.e., a generalized Barker code exists. The analytical results are confirmed by simulations. PMID- 11863494 TI - Nonequilibrium steady states of phonon-fermion systems. AB - Model kinetic equations are used to investigate a nonequilibrium steady state of a phonon-fermion system in the presence of external scatterers. The general relation between the thermodynamic values ensuring a steady state of a fermion gas in the prepense of macroscopic scatterers is found. The effect due to phonon drag is investigated. PMID- 11863495 TI - Particle motion in gas-fluidized granular systems by pulsed-field gradient nuclear magnetic resonance. AB - Nuclear magnetic resonance k-space and q-space imaging methods were applied for the first time to monitor the time-averaged density variations and the random motion of granular particles in the presence of gas flow of variable velocity in a model gas-fluidized bed reactor. The transitions toward fluidization, bubbling phase, and slugging phase are visible as sudden changes in the density and the root mean-squared displacement. The onset of these transitions is shown to depend on the particle sizes for a given cell geometry. The effective diffusion coefficients are measured and the distributions of particle displacements, or propagators, are presented. PMID- 11863496 TI - Frenet algorithm for simulations of fluctuating continuous elastic filaments. AB - We present an algorithm for generating the equilibrium configurations of fluctuating continuous elastic filaments, based on a combination of statistical mechanics and differential geometry. We use this to calculate the distribution function of the end-to-end distance of filaments with nonvanishing spontaneous curvature and show that for small twist and large bending rigidities there is an intermediate temperature range in which the filament becomes nearly completely stretched. We show that volume interactions can be incorporated into our algorithm, demonstrating this through the calculation of the effect of excluded volume on the end-to-end distance of the filament. PMID- 11863497 TI - Rotational Brownian motion and nonlinear dielectric relaxation of asymmetric top molecules in strong electric fields. AB - A general theoretical treatment of the nonlinear dielectric response of an assembly of asymmetric top molecules in strong electric fields is presented in the context of the noninertial rotational diffusion model. The calculation proceeds by obtaining an infinite hierarchy of recurrence equations for the expectation values of Wigner's D functions describing nonlinear relaxation of the system. This hierarchy may be used for the evaluation of both transient and ac nonlinear responses in strong electric fields. The solution of this hierarchy is obtained for the particular case of rigid rodlike molecules in superimposed ac and strong dc bias electric fields, allowing one to evaluate the corresponding nonlinear response. The results are in agreement with available experimental data on nonlinear dielectric relaxation of dilute solutions of polar rodlike molecules in nonpolar solvents. PMID- 11863498 TI - Optimizing of recurrence plots for noise reduction. AB - We propose a way to automatically detect the best neighborhood size for a local projective noise reduction filter, where a typical problem is the proper identification of the noise level. Here we make use of concepts from the recurrence quantification analysis in order to adaptively tune the filter along the incoming time series. We define an index, to be computed via recurrence plots, whose minimum gives a clear indication of the best size of the neighborhood in the embedding space. Comparison of the local projective noise reduction filter using this optimization scheme with the state of the art is also provided. PMID- 11863500 TI - Branched growth with eta approximately 4 walkers. AB - Diffusion-limited aggregation has a natural generalization to the "eta models," in which eta random walkers must arrive at a point on the cluster surface in order for growth to occur. It has recently been proposed that in spatial dimensionality d=2, there is an upper critical eta(c)=4 above which the fractal dimensionality of the clusters is D=1. I compute the first-order correction to D for eta<4, obtaining D=1 + 1/2 (4-eta). The methods used can also determine multifractal dimensions to first order in 4-eta. PMID- 11863499 TI - Generalized Taylor-Aris dispersion in discrete spatially periodic networks: microfluidic applications. AB - A theory is presented for the lumped parameter, convective-diffusive transport of individual, noninteracting Brownian solute particles ("macromolecules") moving within spatially periodic, solvent-filled networks--the latter representing models of chip-based microfluidic chromatographic separation devices, as well as porous media. Using graph-theoretical techniques, the composite medium is conceptually decomposed into a network of channels (the edges) through which the solute is transported by a combination of molecular diffusion and either "piggyback" entrainment within a flowing solvent or an externally applied force field acting upon the solute molecules. A probabilistic choice of egress channel for a solute particle exiting the intersection (vertex) of the channels is furnished by an imperfect mixing model. A spatially periodic, Taylor-Aris-like "method-of-moments" scheme is applied to this transport model, leading to discrete matrix equations for computing the network-scale particle velocity vector U(*) and dispersivity dyadic D(*) in terms of the prescribed microscale transport parameters and network geometry characterizing the basic unit cell of which the spatially periodic device is comprised. The ensuing algebraic equations governing the vertex-based, discrete unit-cell "fields" P(0)(infinity)(i) and B(i) (i=1,2,...,n), whose paradigmatic summations yield U(*) and D(*), constitute discrete analogs of classical continuous macrotransport phenomenological parameters, P(0)(infinity)(r) and B(r), with r a continuous position vector defined within the unit cell. The ease with which these discrete calculations can be performed for complex networks renders feasible parametric studies of potential microfluidic chip designs, particularly those pertinent to biomolecular separation schemes. Application of this discrete theory to the dispersion analysis of pressure-driven flow in spatially periodic serpentine microchannels is shown to accord with existing results previously derived using classical continuous macrotransport theory. PMID- 11863501 TI - Analytic expression for the mean time to absorption for a random walker on the Sierpinski gasket. AB - The exact analytic expression for the mean time to absorption (or mean walk length) for a particle performing a random walk on a finite Sierpinski gasket with a trap at one vertex is found to be T((n))=[3(n)5(n+1)+4(5(n)) 3(n)]/(3(n+1)+1) where n denotes the generation index of the gasket, and the mean is over a set of starting points of the walk distributed uniformly over all the other sites of the gasket. In terms of the number N(n) of sites on the gasket and the spectral dimension d of the gasket, the precise asymptotic behavior for large N(n) is T((n))-->1/3(2N(n))(2/d)-N1.464. This serves as a partial check on our result, as it is (a) intermediate between the known results T-N2 (d=1) and T-N ln N (d=2) for random walks on d-dimensional Euclidean lattices and (b) consistent with the known result for the asymptotic behavior of the mean number of distinct sites visited in a random walk on a fractal lattice. PMID- 11863502 TI - Free random Levy matrices. AB - Using the theory of free random variables and the Coulomb gas analogy, we construct stable random matrix ensembles that are random matrix generalizations of the classical one-dimensional stable Levy distributions. We show that the resolvents for the corresponding matrices obey transcendental equations in the large size limit. We solve these equations in a number of cases, and show that the eigenvalue distributions exhibit Levy tails. For the analytically known Levy measures we explicitly construct the density of states using the method of orthogonal polynomials. We show that the Levy tail distributions are characterized by a different novel form of microscopic universality. PMID- 11863503 TI - Self-avoiding walks on Sierpinski lattices in two and three dimensions. AB - The scaling properties of linear polymers on deterministic fractal structures, modeled by self-avoiding random walks (SAW) on Sierpinski lattices in two and three dimensions, are studied. To this end, all possible SAW configurations of N steps are enumerated exactly and averages over suitable sets of starting lattice points for the walks are performed to extract the mean quantities of interest reliably. We determine the critical exponent describing the mean end-to-end chemical distance (-)l(N) after N steps and the corresponding distribution function, P(S)(l,N). A des Cloizeaux-type relation between the exponent characterizing the asymptotic shape of the distribution, for l-->0 and N--> infinity, and the one describing the total number of SAW of N steps is suggested and supported by numerical results. These results are confronted with those obtained recently on the backbone of the incipient percolation cluster, where the corresponding exponents are very well described by a generalized des Cloizeaux relation valid for statistically self-similar structures. PMID- 11863504 TI - Closed solution to the Baker-Campbell-Hausdorff problem: exact effective Hamiltonian theory for analysis of nuclear-magnetic-resonance experiments. AB - A closed solution to the Baker-Campbell-Hausdorff problem is described. The solution, which is based on the Cayley-Hamilton theorem, allows the entanglement between exponential operators to be described by an exact finite series expansion. Addressing specifically the special unitary Lie groups SU(2), SU(3), and SU(4), we derive expansion formulas for the entangled exponential operator as well as for the effective Hamiltonian describing the net evolution of the quantum system. The capability of our so-called exact effective Hamiltonian theory for analytical and numerical analysis is demonstrated by evaluation of multiple-pulse methods within liquid- and solid-state nuclear-magnetic-resonance spectroscopy. The examples include composite pulses for inversion, decoupling, and dipolar recoupling, as well as coherence-order- and spin-state-selective double- to single-quantum conversion, homonuclear dipolar decoupling, finite rf excitation for quadrupolar nuclei, heteronuclear coherence transfer, and gates for quantum computation. PMID- 11863506 TI - van der Waals equation of state for a fluid in a nanopore. AB - A generalization of the van der Waals equation of state is presented for a confined fluid in a nanopore. The pressure in the fluid, confined in a narrow pore of infinite length, has tensorial character. From this hypothesis, the Helmholtz free energy is constructed and expressions for the axial and transversal components of the pressure tensor are obtained. The equations predict liquid-vapor equilibria, and a shift of the critical point with respect to that obtained from the van der Waals bulk equation. The results are in good agreement with recent experiments. PMID- 11863505 TI - Approach to quantum Kramers' equation and barrier crossing dynamics. AB - We have presented a simple approach to quantum theory of Brownian motion and barrier crossing dynamics. Based on an initial coherent state representation of bath oscillators and an equilibrium canonical distribution of quantum-mechanical mean values of their co-ordinates and momenta we have derived a c number generalized quantum Langevin equation. The approach allows us to implement the method of classical non-Markovian Brownian motion to realize an exact generalized non-Markovian quantum Kramers' equation. The equation is valid for arbitrary temperature and friction. We have solved this equation in the spatial diffusion limited regime to derive quantum Kramers' rate of barrier crossing and analyze its variation as a function of the temperature and friction. While almost all the earlier theories rest on quasiprobability distribution functions (e.g., Wigner function) and path integral methods, the present work is based on true probability distribution functions and is independent of path integral techniques. The theory is a natural extension of the classical theory to quantum domain and provides a unified description of thermally activated processes and tunneling. PMID- 11863507 TI - Entropy production and excess entropy in a nonequilibrium steady-state of single macromolecules. AB - Based on a recently developed formalism for mesoscopic stochastic dynamics of single macromolecules, such as motor proteins, in aqueous solution, we demonstrate mathematically the principle of the nonequilibrium thermodynamics originated by the Brussels group. The key concepts of excess entropy and excess entropy production, and their mathematical properties as well as physical interpretations, are discussed. The newly developed stochastic macromolecular mechanics is consistent with the general theory of nonequilibrium thermodynamics far from equilibrium, and more importantly, it bridges the abstract theory with the current experimental and modeling work on molecular motors and other biological systems in nonequilibrium steady state. PMID- 11863508 TI - Diffusion on random-site percolation clusters: theory and NMR microscopy experiments with model objects. AB - Quasi-two-dimensional random-site percolation model objects were fabricated based on computer-generated templates. Samples consisting of two compartments, a reservoir of H2O gel attached to a percolation model object, which was initially filled with D2O, were examined with nuclear magnetic resonance microscopy for rendering proton spin density maps. The propagating proton/deuteron interdiffusion profiles were recorded and evaluated with respect to anomalous diffusion parameters. The deviation of the concentration profiles from those expected for unobstructed diffusion directly reflects the anomaly of the propagator for diffusion on a percolation cluster. The fractal dimension of the random walk d(w) evaluated from the diffusion measurements on the one hand and the fractal dimension d(f) deduced from the spin density map of the percolation object on the other permits one to experimentally compare dynamical and static exponents. Approximate calculations of the propagator are given on the basis of the fractional diffusion equation. Furthermore, the ordinary diffusion equation was solved numerically for the corresponding initial and boundary conditions for comparison. The anomalous diffusion constant was evaluated and is compared to the Brownian case. Some ad hoc correction of the propagator is shown to pay tribute to the finiteness of the system. In this way, anomalous solutions of the fractional diffusion equation could experimentally be verified. PMID- 11863509 TI - Non-Gaussian equilibrium distributions arising from the Langevin equation. AB - We study the Langevin equation of a point particle driven by random noise, modeled as a two-state Markov process. The corresponding master equation differs from the Fokker-Planck equation. In equilibrium, the velocity of the particle is distributed according to a binomial power law. We discuss transient (i.e., nonequilibrium) behavior, and the consequences of non-Markovian noise statistics. PMID- 11863510 TI - Approximate symmetry laws for percolation in complex systems: Percolation in polydisperse composites. AB - The concept of so-called global symmetry of percolation models is discussed and extended to multicolored models. An integral equation is obtained, which determines the partial percolation probabilities P(a) for sites of color a. This equation is applied to a polydisperse particulate composite: a mixture of conducting (of relative fraction x(m)) and nonconducting spheres with distributions of sizes n(m)(R) and n(i)(R), respectively. We find the probability P(R) for a conducting particle of radius R to belong to the percolation cluster as a function of x(m) and a functional of n(m)(R') and n(i)(R'). The percolation threshold x is shown to decrease with increasing dispersion Delta of particle sizes. A simple law x=1/(3[1+(Delta/4)]) is obtained in the range of moderate dispersions. The theory is applicable also to a mixture of electronic and ionic conductors. PMID- 11863511 TI - Symmetry-breaking instability and strongly peaked periodic clustering states in a driven granular gas. AB - An ensemble of inelastically colliding grains driven by a horizontally vibrating wall in two dimensions exhibits clustering. Working in the limit of nearly elastic collisions and employing granular hydrodynamics, we predict, by a marginal stability analysis, a spontaneous symmetry breaking of the laterally uniform clustering state. Two-dimensional steady-state solutions found numerically describe laterally periodic clustering states. Well within the instability region the density of these states is strongly peaked, with most of the granulate located in "density islands." Time-dependent granular hydrodynamic simulations show that strongly peaked states can develop from small-amplitude single-mode density perturbations. PMID- 11863512 TI - Anomalous behavior of normal kinematic restitution in the oblique impacts of a hard sphere on an elastoplastic plate. AB - We observe oblique impacts of a hard aluminum oxide sphere on a thick elastoplastic polycarbonate plate by recording stroboscopic photographs of the sphere trajectory and spin. The apparent kinematic coefficient of normal restitution grows monotonically with the magnitude of the tangent of the angle of incidence, and the apparent coefficient of friction decreases with increasing normal impact velocity. Although every collision dissipates the total kinetic energy of the sphere, we observe restitution coefficients exceeding unity for the most grazing impacts. We exploit this example to confirm that, although an apparent kinematic coefficient of normal restitution below one is sufficient to guarantee dissipation of kinetic energy in any collision, this condition is not necessary for oblique impacts of spheres on a plate. PMID- 11863513 TI - Dynamics of rarefied granular gases. AB - This paper presents quite general bidimensional gas-dynamic equations--derived from kinetic theory-which include the fourth cumulant kappa(r,t) as a dynamic field. The dynamics describes a low-density system of inelastic hard spheres (disks) with normal restitution coefficient r. Two illustrative examples are given and the role of kappa in them is discussed. Our general gas-dynamic equations would deal with 9 hydrodynamic fields (which corresponds to 14 in three dimension). These fields are the standard hydrodynamic fields plus the components p(ij) of the traceless part of the pressure tensor, the energy flux vector Q and the fourth cumulant kappa. The present formulation requires no constitutive equations. The two examples are: the well-known homogeneous cooling state and a system, with and without gravity, steadily heated by two parallel walls. In the first case, the dynamics yield a description of the homogeneous cooling state consistent with known results adding extra details mainly about the transient time behavior. The steadily heated system kept in a static state gives rise to quite simple but nontrivial equations. In the case with gravity, it is shown that when kappa is included as a dynamic field, the formalism leads to a non-Fourier law already to first order in dissipation. Setting gravity g=0 a perturbative solution is shown and favorably compared with observations obtained from molecular dynamics (MD). In both cases, with and without gravity, kappa is not homogeneous. An analytic extension suggests a divergent situation for a small negative value of q, which originates in the unavoidable extension of the formalism to exothermic collisions associated with a restitution coefficient larger than one. This divergent behavior is observed in MD. PMID- 11863514 TI - Connections between response modes in a horizontally driven granular material. AB - The behavior of a horizontally vibrated quasi-two-dimensional granular system is observed over a wide range of time scales by mapping the velocity fields at the boundary by using high-speed video and decomposing the behavior of the system into the harmonic, subharmonic, and convective responses. The observed relationships between these responses, as well as the fast shearing and the gap that opens between the material and the sidewalls, lead to a refinement of the current convection model, and also reveal that shearing is completely accounted for by the harmonic response. We find that internal degrees of freedom are always significant under horizontal vibration, as the specific boundary conditions (open surface and horizontal driving) give rise to a surface layer whose motion is decoupled by a narrow shear band from that of the bulk. All observed responses are related to this shear band. For example, the specific shape of the shear band gives rise to the surprising presence of period doubling and period quadrupling. As compared to the case of vertical excitation, all the observed responses now arise through new mechanisms. PMID- 11863515 TI - Random-walk simulations of NMR dephasing effects due to uniform magnetic-field gradients in a pore. AB - A random-walk simulation program was developed to study the effect of dephasing spins in a uniform magnetic-field gradient in a porous material. It is shown that this simulation program correctly reproduces basic nuclear magnetic resonance behavior, such as the formation of a spin echo. The spin-echo decay due to dephasing in a nonrestricted medium gives the well-known exponential relation containing the cube of time, whereas the spin-echo decay due to dephasing in a porous material gives a monoexponential decay. By varying the pore size and magnetic-field gradient, the motional averaging regime and the localization regime can be simulated. Moreover, the unknown intermediate regime is investigated. By choosing the right scaling parameters, the spin-echo decay due to dephasing in a pore can be described by one master curve for all pore sizes and gradient strengths. This master curve reveals a small intermediate regime, perfectly symmetrical around the gradient for which the dephasing length is exactly equal to the structural length of the pore. PMID- 11863516 TI - Pressure dependence of the sound velocity in a two-dimensional lattice of Hertz Mindlin balls: mean-field description. AB - We study the dependence on the external pressure P of the velocities v(L,T) of long wavelength sound waves in a confined two-dimensional hexagonal close-packed lattice of 3D elastic frictional balls interacting via one-sided Hertz-Mindlin contact forces, whose diameters exhibit mild dispersion. The presence of an underlying long range order enables us to build an effective medium description, which incorporates the radial fluctuations of the contact forces acting on a single site. Due to the nonlinearity of Hertz elasticity, self-consistency results in a highly nonlinear differential equation for the "equation of state" linking the effective stiffness of the array with the applied pressure, from which sound velocities are then obtained. The results are in excellent agreement with existing experimental results and simulations in the high- and intermediate pressure regimes. It emerges from the analysis that the departure of v(L)(P) from the ideal P(1/6) Hertz behavior must be attributed primarily to the fluctuations of the stress field, rather than to the pressure dependence of the number of contacts. PMID- 11863517 TI - Electrorotation of a pair of spherical particles. AB - We present a theoretical study of electrorotation (ER) of two spherical particles under the action of a rotating electric field. When the two particles approach and finally touch, the mutual polarization interaction between the particles leads to a change in the dipole moment of the individual particle and hence the ER spectrum, as compared to that of the well-separated particles. The mutual polarization effects are captured by the method of multiple images. From the theoretical analysis, we find that the mutual polarization effects can change the characteristic frequency at which the maximum angular velocity of electrorotation occurs. The numerical results can be understood in the spectral representation theory. PMID- 11863518 TI - Lane formation in colloidal mixtures driven by an external field. AB - The influence of an external field on a binary colloidal mixture performing Brownian dynamics in a solvent is investigated by nonequilibrium computer simulations and simple theory. In our model, one half of the particles are pushed into the field direction while the other half of them are pulled into the opposite direction. For increasing field strength, we show that the system undergoes a nonequilibrium phase transition from a disordered state to a state characterized by lane formation parallel to the field direction. The lanes are formed by the same kind of particles moving collectively with the field. Lane formation accelerates particle transport parallel to the field direction but suppresses massively transport perpendicular to the field. We further show that lane formation also occurs in a time-dependent oscillatory field. If the frequency of the external field exceeds a critical value, however, the system exhibits a transition back to the disordered state. Our results can be experimentally verified in binary colloidal suspensions exposed to external fields under nonequilibrium conditions. PMID- 11863519 TI - Supersensitivity in a chain of closely spaced electric dipoles with variable moments. AB - A chain of closely spaced oscillators is studied theoretically. The oscillators are interrelated electric dipoles whose moments may vary within a wide range. An expression for the oscillator interaction potential is suggested. On the basis of this potential, a one-dimensional nonlinear equation of motion is derived with allowance made for dissipation and external driving. A numerical investigation is carried out, and various nonlinear phenomena are revealed in the chain. Among them are the size effect and ultrasensitivity, i.e., a giant response of the chain to extremely weak periodic perturbations. The findings are compared with previously obtained experimental results on naturally occurring objects with similar structure. It is inferred that the model is realistic. PMID- 11863520 TI - Cell growth in a pinned soap froth. AB - Experimental kinetic data on soap froth cell growth in a random array of fixed pinning centers is reported. The scaling of our two-dimensional data is analyzed and compared with pinning-free growth. The obtained data agree fairly well with a proposed cell growth law. We also discuss the relationship between the pin concentration and the final cell size. PMID- 11863521 TI - Adsorption of colloidal particles on a charged surface: cluster Monte Carlo simulations. AB - We investigate the adsorption of colloidal particles on a charged surface by the cluster Monte Carlo simulation. Under the primitive model of asymmetric electrolytes, the density profiles of colloidal particles at a surface are analyzed as a function of the electrostatic coupling parameter. In the strong coupling regime numerical results show the like-charged adsorption between colloidal particles and a charged surface, which cannot be explained by the Derjaguin-Landau-Verwey-Overbeek theory. PMID- 11863522 TI - Short-time dynamics of colloidal particles confined between two walls. AB - The short-time dynamics of colloidal particles in a quasi-two-dimensional geometry is studied by digital video microscopy. The particles (polystyrene spheres) are suspended in water and confined between two parallel glass plates, forming an effective two-dimensional system. The (effective) two-dimensional van Hove function G(r,t) and its self and distinct part are measured with a time resolution of 1/30 s. We found that the general behavior of these time correlation functions (and their Fourier transforms) is quite similar to that of their three-dimensional counterparts. The effects of the strong hydrodynamic coupling of the particles motion to the walls and that due to the hydrodynamic interactions between particles are contained in the (effective) hydrodynamic function H(k) obtained from the initial slope of F(k,t) [the Fourier transform of G(r,t)]. We found that H(k), as a function of the wave vector k and particle concentration, exhibits a similar qualitative behavior to the hydrodynamic function in homogeneous three-dimensional suspensions of hard spheres. We also found in our systems that the particle fluctuations relax only by self-diffusion for wave vectors where the static structure factor S(k)=1. This result is important for measurements of self-diffusion dynamics in three-dimensional systems by light scattering techniques. PMID- 11863523 TI - Viscosity and transient electric birefringence study of clay colloidal aggregation. AB - We study a synthetic clay suspension of laponite at different particle and NaCl concentrations by measuring stationary shear viscosity and transient electrically induced birefringence (TEB). On one hand the viscosity data are consistent with the particles being spheres and the particles being associated with large amount bound water. On the other hand the viscosity data are also consistent with the particles being asymmetric, consistent with single laponite platelets associated with a very few monolayers of water. We analyze the TEB data by employing two different models of aggregate size (effective hydrodynamic radius) distribution: (1) bidisperse model and (2) log-normal distributed model. Both models fit, in the same manner, fairly well to the experimental TEB data and they indicate that the suspension consists of polydisperse particles. The models also appear to confirm that the aggregates increase in size vs increasing ionic strength. The smallest particles at low salt concentrations seem to be monomers and oligomers. PMID- 11863524 TI - Do liquids exhibit local fivefold symmetry at interfaces? AB - We calculate the layer-resolved local fivefold symmetry of liquids near interfaces using computer simulation of hard sphere fluids on structured substrates. In the first few adjacent liquid layers, the presence of a surface suppresses local icosahedral packing while the magnitude of local fivefold symmetry in the next layers depends on details of the particle-substrate interaction. For a strong particle-substrate attraction, the local fivefold symmetry in these layers is higher than in the bulk liquid. PMID- 11863525 TI - Phase diagram and physical properties of a waterless sodium bis(2 ethylhexylsulfosuccinate)- ethylbenzene-ethyleneglycol microemulsion: an insight into percolation. AB - Volumetric and transport studies have been carried out for the nonaqueous ternary microemulsion system containing sodium bis(2-ethylhexylsulfosuccinate) aerosol-OT (AOT), ethylbenzene (EB), and ethyleneglycol (EG). The results obtained for the conductivity sigma are presented over a wide range of volume fraction of dispersed phase phi and different molar concentration ratio omega=[EG]/[AOT]=2-10 at 30 degrees C and discussed in context of percolation theory. The variation of sigma with respect to temperature (T=10-60 degrees C) shows an increase in the conductance values but no percolation-type phenomenon is observed. The measurements of viscosity, density, and ultrasonic velocity have also been carried out to understand the behavior of this nonaqueous microemulsion system. The phase behavior of the microemulsion is sensitively dependent on the EG to AOT molar ratio. A simple structural model has been applied for the calculation of the various parameters, i.e., aggregation number (n), core radius (r(n)), and surface number density of the surfactant molecule at interface (alpha(s)). PMID- 11863526 TI - Interfaces of polydisperse fluids: surface tension and adsorption properties. AB - We consider a system of spherical colloidal particles with a size polydispersity and use a simple van der Waals description in order to study the combined effect of both the polydispersity and the spatial nonuniformity induced by a planar interface between a low-density fluid phase (enriched in small particles) and a high-density fluid phase (enriched in large particles). We find a strong adsorption of small particles at the interface, the latter being broadened with respect to the monodisperse case. We also find that the surface tension of the polydisperse system results from a competition between the tendency of the polydispersity to lower the surface tension and its tendency to raise the critical-point temperature (i.e., its tendency to favor phase separation) with the former tendency winning at low temperatures and the latter at the higher temperatures. PMID- 11863527 TI - Dynamics of nanoscale droplets. AB - Several aspects of the dynamical behavior of nanometer-sized liquid drops have been investigated using molecular dynamics computer simulations. The liquids consist of chains of lengths 2-100 with Lennard-Jones and nonlinear elastic interactions, comprising drops made of O(100,000) atoms with radius O(10 nm), placed on an atomistic substrate. We simulate the solidification of a liquid drop on a substrate as the latter is cooled, and observe a smooth contraction and smoothing process but no dimple formation. If instead we simulate heating of the substrate, the drop evaporates, and we find a decrease on contact angle in the partially wetting case, in accord with recent predictions. Last, we consider the coalescence of two such droplets, either spontaneously or when pushed together, to study shape evolution scaling laws and to document a rapid internal structural reequilibration. Aside from the intrinsic novelty associated with the small scale of these systems, the results illustrate the relation between atomic scale interactions and macroscopic continuum properties of materials. PMID- 11863528 TI - Fluids of platelike particles near a hard wall. AB - Fluids consisting of hard platelike particles near a hard wall are investigated using density-functional theory. The density and orientational profiles, as well as the surface tension and the excess coverage, are determined and compared with those of a fluid of rodlike particles. Even for low densities, slight orientational packing effects are found for the platelet fluid due to larger intermolecular interactions between platelets as compared with those between rods. A net depletion of platelets near the wall is exhibited by the excess coverage, whereas a change of sign of the excess coverage of hard-rod fluids is found upon increasing the bulk density. PMID- 11863529 TI - Kob-Andersen model: a nonstandard mechanism for the glassy transition. AB - We present results reflecting the analogies between the Kob-Andersen model and other glassy systems. Studying the stability of the blocked configurations above and below the transition we also give arguments that support their relevance for the glassy behavior of the model. However, we find, surprisingly, that the organization of the phase space of the system is different from the well known organization of other mean field spin glasses and structural glasses. PMID- 11863530 TI - Reduction of the glass transition temperature in polymer films: a molecular dynamics study. AB - We present results of molecular-dynamics simulations for a nonentangled polymer melt confined between two completely smooth and repulsive walls, interacting with inner particles via the potential U(wall)=(sigma/z)(9), where z=/z(particle) z(wall) and sigma is (roughly) the monomer diameter. The influence of this confinement on the dynamic behavior of the melt is studied for various film thicknesses (wall-to-wall separations) D, ranging from about 3 to about 14 times the bulk radius of gyration. A comparison of the mean-square displacements in the film and in the bulk shows an acceleration of the dynamics due to the presence of the walls. This leads to a reduction of the critical temperature T(c) of the mode coupling theory with decreasing film thickness. Analyzing the same data by the Vogel-Fulcher-Tammann (VFT) equation, we also estimate the VFT temperature T0(D). The ratio T0(D)/T(bulk)(0) decreases for smaller D similarly to T(c)(D)/T(bulk)(c). These results are in qualitative agreement with that of the glass transition temperature observed in some experiments on supported polymer films. PMID- 11863531 TI - Colloids, polymers, and needles: demixing phase behavior. AB - We consider a ternary mixture of hard colloidal spheres, ideal polymer spheres, and rigid vanishingly thin needles, which model stretched polymers or colloidal rods. For this model, we develop a geometry-based density functional theory, apply it to bulk fluid phases, and predict demixing phase behavior. In the case of no polymer-needle interactions, two-phase coexistence between colloid-rich and colloid-poor phases is found. For hard needle-polymer interactions, we predict rich phase diagrams, exhibiting three-phase coexistence, and reentrant demixing behavior. PMID- 11863532 TI - Critical concentration fluctuations of the ionic binary mixture ethylammonium nitrate-n-octanol: an ultrasonic spectrometry study. AB - Between 200 kHz and 130 MHz, the ultrasonic attenuation spectrum of the ionic ethylammonium nitrate--n-octanol mixture of critical composition has been measured at various reduced temperatures (1.5 x 10(-4)0) behavior of the product of the vibrational density of states and the Raman coupling coefficient has also been extracted. The validity of some phenomenological approaches is also discussed in light of the experimental facts presented in this paper and some suggestions are being advanced. PMID- 11863534 TI - Fractal analysis of sampled profiles: systematic study. AB - A quantitative evaluation of the influence of sampling on the numerical fractal analysis of experimental profiles is of critical importance. Although this aspect has been widely recognized, a systematic analysis of the sampling influence is still lacking. Here we present the results of a systematic analysis of synthetic self-affine profiles in order to clarify the consequences of the application of a poor sampling (up to 1000 points) typical of scanning probe microscopy for the characterization of real interfaces and surfaces. We interpret our results in terms of a deviation and a dispersion of the measured exponent with respect to the "true" one. Both the deviation and the dispersion have always been disregarded in the experimental literature, and this can be very misleading if results obtained from poorly sampled images are presented. We provide reasonable arguments to assess the universality of these effects and propose an empirical method to take them into account. We show that it is possible to correct the deviation of the measured Hurst exponent from the "true" one and give a reasonable estimate of the dispersion error. The last estimate is particularly important in the experimental results since it is an intrinsic error that depends only on the number of sampling points and can easily overwhelm the statistical error. Finally, we test our empirical method calculating the Hurst exponent for the well-known 1+1 dimensional directed percolation profiles, with a 512-point sampling. PMID- 11863535 TI - Fingering instability of thin evaporating liquid films. AB - The fingering instability of growing dry patches in an evaporating film of a polar liquid placed on a solid substrate is investigated. The instability manifests itself as fingering of mobile fronts between growing "dry" (thin) and shrinking "wet" (thick) regions of the film corresponding to two stable states of the evaporating film in contact with its vapor. The boundaries of the fingering instability are found through linear stability analysis of numerical solutions of the nonlinear evolution equation defining the film profile, and the influence of the evaporation rate, polar intermolecular forces, and chemical heterogeneity of the substrate is investigated. PMID- 11863536 TI - Wetting droplet instability and quantum ring formation. AB - InAs islands on GaAs substrates undergo a morphological change into ring-shaped configurations upon deposition of a GaAs layer after island growth. By invoking an analogy of the InAs islands to wetting droplets on solid substrates, we suggest that this transition might be brought about by a change of the surface free-energy balance at the three-phase contact-line between GaAs, InAs, and vacuum (or As atmosphere). Our scenario can also be tested in conventional liquid systems (e.g., polymers). PMID- 11863537 TI - Dynamical fluctuations in ion conducting glasses: slow and fast components in lithium metasilicate. AB - Molecular dynamics simulations of lithium metasilicate (Li2SiO3) glass have been performed. Dynamic heterogeneity of lithium ions has been examined in detail over 4 ns at 700 K. Type A particles show slow dynamics in accordance with a long tail of waiting time distribution of jump motion and localized jumps within neighboring sites (fracton), while type B particles show fast dynamics related to cooperative jumps and a strong forward correlated motion (Levy flight). Mutual changes of two kinds of dynamics with the relatively long time scale have been observed. The changes cause an extremely large fluctuation of the mean squared displacements as well as the squared displacement of each particle, which depends on the time window of observation. Localized jump motion (fracton) cannot contribute to the long-time-translational diffusion but it can contribute to the rotational diffusion. On the other hand, forward correlated jump motion mainly contributes to long-time-translational diffusion and not to the short-time rotational diffusion, although this can be a slower part of the rotational diffusion. These results have been compared with those of simple glass-forming liquids exhibiting the dynamic heterogeneity near T(g). The translation-rotation paradox can be explained by the characteristics of slow and fast dynamics. PMID- 11863538 TI - Wetting and structure of a fluid in a spherical cavity. AB - The equilibrium local densities, structure, and wetting of a one-component fluid in a spherical cavity, of variable radius R, are determined, using density functional theory, as functions of two parameters characterizing the system: the radius R and the cavity/fluid potential parameter epsilon(W). The cavity acts as an external potential V(ext)(r) on the molecules of the confined fluid, the particles of which are of constant diameter d. The equilibrium density profile, as a result of strong confinement, develops peaks in the center of the cavity and/or close to the pore wall and, in certain situations, in other intermediate points; the cavity can also be liquid full, capillary condensation. PMID- 11863539 TI - Morphology and orientational order of nematic liquid crystal droplets confined in a polymer matrix. AB - The orientational order of nematic liquid crystal (LC) droplets confined in a polymer matrix is investigated by 13C-NMR. Different morphologies exhibiting various cavity sizes were obtained by changing the LC fraction. The phase diagram has been established by polarized optical microscopy exhibiting a typical upper critical solution temperature shape. The nematic order increases with decreasing droplet size which may result from increased polymer/LC surface contacts in smaller cavities. The nematic fraction of 4-n-pentyl-4'-cyanobiphenyl (5CB) contained in the droplets was calculated by integrating the NMR peaks, then fitted using a simple theoretical model and compared with differential scanning calorimetry results. NMR investigations in the aliphatic region of the spectra have shown that the 2-ethyl hexyl fragment of the polymer chain is partially ordered at the polymer/LC interface due to interdigitation with 5CB molecules. Above the nematic-isotropic transition temperature, a weak pretransitional behavior is shown by the polymer. PMID- 11863540 TI - Two experimental tests of a fluctuation-induced first-order phase transition: intensity fluctuation microscopy at the nematic-smectic-A transition. AB - We have developed a new, extremely sensitive real-space technique (intensity fluctuation microscopy) to probe the order of the nematic-smectic-A (NA) transition. Using this technique, we show that the NA transition in 4'-n-octyl-4 cyanobiphenyl (8CB) is clearly first order, contrary to calorimetric studies but in agreement with conclusions drawn from front-velocity measurements. We characterize the strength of the discontinuity at the first-order transition by the dimensionless quantity t(0)=(T(NA)-T*)/T(*). By precisely measuring t(0), we have made the first detailed tests of predictions based on the Halperin-Lubensky Ma (HLM) theory of fluctuation-induced, first-order phase transitions. First, we explore the effect of an external magnetic field on the NA transition. Although modest fields (of order 10 T) are predicted to drive the weakly first-order transition in pure 8CB second order, we observe no such effect; we establish instead that the lower bound on this critical field is approximately 30 T. Likewise, we observe no effect in mixtures of 8CB with its longer chemical homolog 4'-n-decyl-4-cyanobiphenyl (10CB). Second, we examine the dependence of t(0) as a function of 8CB-10CB mixture concentration and find that the data in mixtures with small nematic temperature range are well-fit by the parameters derived by Anisimov et al. based on calorimetric measurements. As we increase the nematic range (by using concentrations closer to pure 8CB), the measured t(0) deviates more and more from the HLM predictions. Smectic fluctuations, which are neglected in the HLM calculation, are an obvious candidate to explain such a discrepancy, but one's naive expectation is that they would reduce t(0) below the HLM levels, whereas the observed values are too large. However, a recent renormalization-group calculation concludes that smectic fluctuations, surprisingly, should indeed increase t(0), explaining the observations presented here. PMID- 11863541 TI - Surface-induced nematic and smectic order at a liquid-crystal-silanated-glass interface observed by atomic force spectroscopy and Brewster angle ellipsometry. AB - We have investigated nematic and smectic surface-induced ordering in the isotropic phase of the 4-cyano-4'-n-octylbiphenyl liquid crystal on silanated BK7 and LaSF glass using atomic force spectroscopy and Brewster angle ellipsometry. We have observed complete wetting of the silanated glass surfaces with the nematic phase when approaching the isotropic-nematic phase transition from above. Using the atomic force spectroscopy at the same interfaces, we have detected a significant presmectic ordering that is enhanced at the nematic-isotropic transition. We have observed the first, strongly adsorbed layer of liquid crystalline molecules underneath this presmectic ordering. This first molecular layer is laterally inhomogeneous with typically 100 nm voids and covers approximately 70% of the surface. It is stable far above the clearing point and is responsible for the surface-memory effect. The results have been analyzed using Landau-de Gennes theory. The surface coupling energies of the nematic and smectic order have been determined, as well as the coupling energy between the nematic and smectic order. PMID- 11863542 TI - Theory and simulation of the nematic zenithal anchoring coefficient. AB - Combining molecular simulation, Onsager theory, and the elastic description of nematic liquid crystals, we study the dependence of the nematic liquid crystal elastic constants and the zenithal surface anchoring coefficient on the value of the bulk order parameter. PMID- 11863543 TI - Molecular dynamics study of the isotropic-nematic quench. AB - Effects of cylindrical and spherical confinement on the kinetics of the isotropic nematic quench is studied numerically. The nematic liquid crystal structure was modeled by a modified induced-dipole--induced-dipole interaction. Molecules were allowed to wander around points of a hexagonal lattice. Brownian molecular dynamics was used in order to access macroscopic time scales. In the bulk we distinguish between the early, domain, and late stage regime. The early regime is characterized by the exponential growth of the nematic uniaxial order parameter. In the domain regime domains are clearly visible and the average nematic domain size xi(d) obeys the dynamical scaling law xi(d)-t(gamma). The late stage evolution is dominated by dynamics of individual defects. In a confined system the qualitative change of the scaling behavior appears when xi(d) becomes comparable to a typical linear dimension R of the confinement. In the confining regime (xi(d)>or=R) the scaling coefficient gamma depends on the details of the confinement and also the final equilibrium nematic structure. The domain growth is well described with the Kibble-Zurek mechanism. PMID- 11863544 TI - Description of dynamic x-ray scattering from freely standing smectic-A films. AB - The consequent description of the x-ray scattering from the free standing smectic A films is suggested. Calculations are based on the discrete model for the film dynamics. The scattering intensity temporal autocorrelation function is obtained within the framework of this approach neglecting the multiple scattering and refraction effects. It is shown that the behavior of this function crucially depends on the film thickness. In particular, in thin films containing less than 10(3) layers the time dependence of has damping oscillation character. This behavior is determined by an acoustic mode that describes the film motion caused by the action of the surface tension. For thick films containing more than 10(4) layers the dynamics of the intensity temporal autocorrelation function is determined either by an acoustic mode or by a wide spectrum of modes depending on the x-ray geometry. In both the cases the autocorrelation function is a relaxation one. The results obtained are compared with the experiments on the coherent soft and hard x-ray scattering. PMID- 11863545 TI - Molecular arrangements and reorientation behavior in a dibenzopyrene-derivative ferroelectric columnar liquid crystal as studied by time-resolved Fourier transform ir spectroscopy. AB - Polarized, time-resolved Fourier-transform infrared spectroscopy was employed to study the orientational order and the reorientation dynamics of a diskotic ferroelectric liquid crystal. In the shear oriented cell the dibenzopyrene derivative forms two different field-dependent columnar phases that show a tripling in the spontaneous polarization. These field-dependent phases are analyzed with respect to the dependence of the infrared absorbance from the polarization plane. In this way it was confirmed that the high-field phase is characterized by a homogeneous orientation of the tilt-plane formed by the core normals n and the column axis N. In contrast, in the low-field phase the columns exhibit several different tilt-planes. The orientational order parameter of the columns is determined. It was also detected that the average orientation of the alkyl tails of the molecules is not lying in the plane of the disklike core. By monitoring the evolution of the infrared bands in the course of the electric field-induced reorientation, we found that the reorientation process is divided into three steps: A fast initial response followed by a slowing down of the reorientation is observed, which then is followed by an acceleration of the reorientation. In the high-field phase the fast initial electrical induced process can be assigned to a rotation of the molecules around the column axis by a few degrees. During the subsequent ferroelectriclike response the molecules rotate around the column axis by approximately 180 degrees. Other models for this switching mechanisms could be excluded. PMID- 11863546 TI - Structure and dynamics of ferroelectric liquid crystal cells exhibiting thresholdless switching. AB - Surface-stabilized ferroelectric liquid crystal cells are usually bistable, with the spontaneous polarization either up or down, perpendicular to the plane of the cell. In materials with high spontaneous polarization the polarization charge self-interaction leads to a monostable structure that exhibits a thresholdless, "V-shaped" switching of the optical response to an applied voltage. With a model that includes the electrostatic self-energy of the spontaneous polarization, we examine the transition from bistablity to monostability as a function of the polar and nonpolar anchoring energies, the liquid crystal elasticity, and the thickness of the alignment layers. Two bistable and one monostable structures are possible. The bistable state is only obtained when the thickness of the insulating alignment layer and the polar anchoring energy are small, or when the spontaneous polarization is small. From linear stability analysis we get analytical expressions for the limits of stability of the structures. Numerical calculations show that in some ranges of the parameters two structures can coexist. We also present the calculation of the polarization fluctuation modes. The lowest one becomes soft at the continuous transitions among the structures and at the limits of stability. PMID- 11863548 TI - Time-resolved infrared spectroscopic study of the switching dynamics of a surface stabilized ferroelectric liquid crystal. AB - The orientation dynamics of a ferroelectric liquid crystal with a naphthalene ring (FLC-3) during the electric-field-induced switching between two surface stabilized states was investigated by means of time-resolved Fourier-transform infrared spectroscopy. Time-resolved infrared spectra of the planar-aligned cell of FLC-3 were measured as a function of the polarization angle ranging from 0 degree to 180 degrees under a rectangular electric field +/-40 V with a 5-kHz repetition rate in the smectic-C* (Sm-C*) phase at 137 degrees C. From these spectra details about the mutual arrangement of different molecular segments at all the delay times in the Sm-C* phase were derived. It was found that the C=O group in the core moiety exhibits a dynamical behavior different from that in the chiral moiety during the electric-field-induced switching between the two surface stabilized states. The most important finding in the present study is that during the electric-field-induced switching the FLC molecule not only rotates around the layer normal, but also revolves around its own long axis. Furthermore, time resolved infrared spectroscopy revealed that each group in the core moiety passes almost simultaneously through the projection of the layer normal in the cell window during the dynamic switching. PMID- 11863547 TI - Symmetry breaking and interaction of colloidal particles in nematic liquid crystals. AB - We propose a general approach to the description of the long-ranged elastic interaction in the nematic colloids, based on the symmetry breaking of the director field. The type of the far-field interaction between particles immersed in a nematic host is determined by the way the symmetry is broken in the near field region around the colloidal particle. This is caused both by the particle's shape and the anchoring at the surface. If the director field near the particle has a set of three symmetry planes, the far-field interaction falls off as d(-5) with d being the distance between particles. If one symmetry plane is absent, a dipolar moment perpendicular to it is allowed and yields dipole-dipole interactions, which decays as d(-3). If both the horizontal and vertical mirror symmetries are broken (it is equivalent to the case when the nonzero torque moment is applied to the particle by the nematic liquid crystal), the particles are shown to attract each other following the Coulomb law. We propose a simple method for the experimental observation of this Coulomb attraction. The behavior of colloid particles in curved director fields is analyzed. Quadrupolar particles with planar anchoring are shown to be attracted toward the regions with high splay deformations, while quadrupoles with homeotropic anchoring are depleted from such regions. When there are many colloidal particles in the nematic solvent, the distortions of the director from all of them are overlapped and lead to the exponential screening in the elastic pair interaction potential. This is a many-body interaction effect. This screening is essential in the real dense colloid systems, such as ferronematics--suspensions of magnetic cylindrical grains in the nematic liquid crystal. External magnetic field induces an elastic Yukawa attraction between them. We apply this attraction to the explanation of the cellular texture in magnetically doped liquid crystals. PMID- 11863549 TI - Characterization of free volume during vulcanization of styrene butadiene rubber by means of positron annihilation lifetime spectroscopy and dynamic mechanical test. AB - An experimental investigation was performed to study the effect on the free volume of the advance of the cross-linking reaction in a copolymer of styrene butadiene rubber by sulfur vulcanization. The dynamic modulus and loss tangent were evaluated over samples cured for different times at 433 K by dynamic mechanical tests over a range of frequencies between 5 and 80 Hz at temperatures between 200 and 300 K. Using the William-Landel-Ferry relationship, master curves were obtained at a reference temperature of 298 K and the coefficients c(0)(1) and c(0)(2) were evaluated. From these parameters the dependence of the free volume on the cure time is obtained. Positron annihilation lifetime spectroscopy was also used to estimate the size and number density of free volume sites in the material. The spectra were analyzed in terms of continuous distributions of free volume size. The results suggest an increase of the lower free volume size when cross linking takes place. Both techniques give similar results for the dependence of free volume on the time of cure of the polymer. PMID- 11863550 TI - Nonuniversal nature of dynamic critical anomaly in polymer solutions. AB - Critical anomaly of viscosity has been studied for ideal polymer solutions, focusing on its dependence on the molecular weight of polymer M(w). According to the conventional understanding that polymer solutions should belong to the same dynamic universal class as classical fluids, the critical exponent of viscosity y(c) should be a universal constant (approximately 0.04). Contrary to this, we find that y(c) significantly decreases with increasing M(w). This unusual behavior can be explained by the dynamic coupling of critical concentration fluctuations with an additional slow viscoelastic mode intrinsic to polymer solutions. Our dynamic light scattering measurements support this picture. PMID- 11863551 TI - Disorder and funneling effects on exciton migration in treelike dendrimers. AB - The center-bound excitonic diffusion on dendrimers subjected to several types of nonhomogeneous funneling potentials is considered. We first study the mean first passage time (MFPT) for diffusion in a linear potential with different types of correlated and uncorrelated random perturbations. Increasing the funneling force, there is a transition from a phase in which the MFPT grows exponentially with the number of generations g to one in which it does so linearly. Overall the disorder slows down the diffusion, but the effect is much more pronounced in the exponential compared to the linear phase. When the disorder gives rise to uncorrelated random forces there is, in addition, a transition as the temperature T is lowered. This is a transition from a high-T regime in which all paths contribute to the MFPT to a low-T regime in which only a few of them do. We further explore the funneling within a realistic nonlinear potential for extended dendrimers in which the dependence of the lowest excitonic energy level on the segment length was derived using the time-dependent Hatree-Fock approximation. Under this potential the MFPT grows initially linearly with g but crosses over, beyond a molecular-specific and T-dependent optimal size, to an exponential increase. Finally we consider geometrical disorder in the form of a small concentration of long connections as in the small world model. Beyond a critical concentration of connections the MFPT decreases significantly and it changes to a power law or to a logarithmic scaling with g, depending on the strength of the funneling force. PMID- 11863552 TI - Effect of cross-linker geometry on dynamic mechanical properties of nematic elastomers. AB - We study three monodomain (single-crystal) nematic elastomer materials, all side chain siloxane polymers with the same mesogenic groups but with different types of cross linking: (i) short flexible siloxane linkage affine to the network backbone, (ii) short flexible aliphatic cross links miscible with mesogenic side chain groups, and (iii) long segments of main-chain nematic polymer. The dynamic mechanical response of these three systems shows a characteristically universal decrease of storage modulus and a corresponding increase of loss factor. This effect of "dynamic soft elasticity" is strongly anisotropic, depending on the nematic director orientation. We examine the important role of the average backbone chain anisotropy r(T)=l(parallel)/l(perpendicular), which is affected by the cross-linking geometry and contributes to the magnitude and frequency dependence of the dynamic anomaly, and discuss possible applications in mechanical damping and polarized acoustic technology. PMID- 11863553 TI - Simulation study of the glass transition temperature in poly(methyl methacrylate). AB - The glass transition in syndiotactic poly (methyl methacrylate) has been studied through atomistic molecular dynamics simulations performed at temperatures in the range from 297 K to 684 K. The mean squared deviations of atoms, monomers, and molecules from their initial positions were analyzed by means of a technique that separates the effects of diffusive motion from the underlying vibrational motion. The diffusive motion shows a novel power-law variation with time, with an exponent that varies continuously from 0.5 below the glass transition temperature T(g) to 1 at high temperatures. The self part of the van Hove correlation functions for both hydrogen atoms and monomers shows structural arrest at the lowest temperature studied. A second peak in the atomic van Hove correlation is attributed to rotation of the CH3 group. PMID- 11863554 TI - Viscoelastic effects in relaxation processes of concentration fluctuations in dynamically asymmetric polymer blends. AB - Relaxation processes of the concentration fluctuations induced by a rapid pressure change were investigated for a dynamically asymmetric polymer blend [deuterated polybutadiene (DPB)/polyisoprene (PI)] with a composition of 50-50 by weight by using time-resolved small-angle neutron scattering. The pressure change was carried out inside the single-phase of the blend with the cell designed for polymeric systems under high pressure and temperature. Time change in the scattered intensity distribution with wave number (q) during the relaxation processes was found to be approximated by Cahn-Hilliard-Cook linearized theory. The theoretical analysis yielded the q dependence of Onsager kinetic coefficient that is characterized by the q(-2) dependence at q(xi)(ve)>1 with the characteristic length xi(ve) (with xi(ve) being the viscoelastic length) being much larger than radius of gyration of DPB or PI. The estimated xi(ve) agrees well with that calculated using the Doi and Onuki theory that takes into account the viscoelastic effects arising from the dynamical asymmetry between the component polymers in the relaxation of concentration fluctuations. PMID- 11863555 TI - Local and segmental dynamics in homopolymer and triblock copolymers with one semicrystalline block. AB - Thermally stimulated depolarization currents, TSDC, experiments have been performed on a series of poly(styrene)-b-poly(butadiene)-b-poly(epsilon caprolactone) triblock copolymers SBC with different proportions of the poly(epsilon-caprolactone) crystallizable block, PCL. The morphology of the segregated microphases varies with the PCL content and has been observed by transmission electron microscopy. The crystallinity of the PCL block is estimated by wide angle x-ray scattering, WAXS. The relaxation times distribution is extracted by a numerical decomposition of the TSDC spectra and it is shown that this distribution is not significantly changed on going from the homopolymer to the triblock copolymer with 16 wt % to 77 wt % of PCL in the original samples. Better segregation of the mesophase structure is reached when the samples are annealed at 413 K and important variations in the TSDC and WAXS spectra are observed as a result of the thermal treatment. For the S09B14C77 triblock copolymer the results obtained can be explained by postulating the existence of a rigid amorphous phase in the PCL block. Such rigid amorphous phase is located between the core-shell cylinders formed by the other blocks [with poly(styrene)(PS) as core and poly(butadiene)(PB) as shell] and is constrained by undulated lamellae of crystalline PCL material. In the case of S35B15C50 triblock copolymer, an important amount of diffuse PS-PCL interphase where the homopolymers are mixed must be present before annealing. The results for the material with the less abundant PCL block are explained as a result of the confinement in nanotubes of PCL surrounded by PB embedded in a vitreous PS matrix. Broadband dielectric experiments on these same materials confirm the results obtained by TSDC spectroscopy. PMID- 11863556 TI - Bulk and interfacial shear thinning of immiscible polymers. AB - Nonequilibrium molecular-dynamics simulations are used to study the shear thinning behavior of immiscible symmetric polymer blends. The phase-separated polymers are subjected to a simple shear flow imposed by moving a wall parallel to the fluid-fluid interface. The viscosity begins to shear thin at much lower rates in the bulk than at the interface. The entire shear-rate dependence of the interfacial viscosity is consistent with a shorter effective chain length s(*) that also describes the width of the interface. This s(*) is independent of chain length N and is a function only of the degree of immiscibility of the two polymers. Changes in polymer conformation are studied as a function of position and shear rate. Shear thinning correlates more closely with a decrease in the component of the radius of gyration along the velocity gradient than with elongation along the flow. At the interface, this contraction of chains is independent of N and consistent with the bulk behavior for chains of length s(*). The distribution of conformational changes along chains is also studied. Central regions begin to stretch at a shear rate that decreases with increasing N, while shear induced changes at the ends of chains are independent of N. PMID- 11863557 TI - Noise-induced effects on period-doubling bifurcation for integrate-and-fire oscillators. AB - This study provides a method for calculating first-order approximations of the Lyapunov exponents of systems with discontinuity in the presence of noise in order to characterize the stability of motions in those systems. This paper in particular illustrates the results of the ways in which noise influences period doubling bifurcation in the parameter space of an integrate-and-fire model with a periodically modulated reset level. For evaluating a stochastic version of period doubling bifurcation, the first-passage-time problem of the Ornstein-Uhlenbeck process is used to define a Markov operator governing the transition of a reset level phase density. The results on the stochastic bifurcation are thus compared with numerical computations of angles and moduli of eigenvalues of the Markov operator that describes the firing properties of the model. PMID- 11863558 TI - Nonequilibrium phase transitions in a model for the origin of life. AB - The requisites for the persistence of small colonies of self-replicating molecules living in a two-dimensional lattice are investigated analytically in the infinite diffusion or mean-field limit and through Monte Carlo simulations in the position-fixed or contact process limit. The molecules are modeled by hypercyclic replicators A that are capable of replicating via binary fission A+E- >2A with production rate s, as well as via catalytically assisted replication 2A+E-->3A with rate c. In addition, a molecule can degrade into its source materials E with rate gamma. In the asymptotic regime, the system can be characterized by the presence (active phase) and the absence (empty phase) of replicators in the lattice. In both diffusion regimes, we find that for small values of the ratio c/gamma these phases are separated by a second-order phase transition that is in the universality class of the directed percolation, while for small values of s/gamma the phase transition is of first order. Furthermore, we show the suitability of the dynamic Monte Carlo method, which is based on the analysis of the spreading behavior of a few active cells in the center of an otherwise infinite empty lattice, to address the problem of the emergence of replicators. Rather surprisingly, we show that this method allows an unambiguous identification of the order of the nonequilibrium phase transition. PMID- 11863559 TI - Generic model of morphological changes in growing colonies of fungi. AB - Fungal colonies are able to exhibit different morphologies depending on the environmental conditions. This allows them to cope with and adapt to external changes. When grown in solid or semisolid media the bulk of the colony is compact and several morphological transitions have been reported to occur as the external conditions are varied. Here we show how a unified simple mathematical model, which includes the effect of the accumulation of toxic metabolites, can account for the morphological changes observed. Our numerical results are in excellent agreement with experiments carried out with the fungus Aspergillus oryzae on solid agar. PMID- 11863560 TI - Percolation and epidemics in a two-dimensional small world. AB - Percolation on two-dimensional small-world networks has been proposed as a model for the spread of plant diseases. In this paper we give an analytic solution of this model using a combination of generating function methods and high-order series expansion. Our solution gives accurate predictions for quantities such as the position of the percolation threshold and the typical size of disease outbreaks as a function of the density of "shortcuts" in the small-world network. Our results agree with scaling hypotheses and numerical simulations for the same model. PMID- 11863561 TI - Flexoelectricity and elasticity of asymmetric biomembranes. AB - In view of the well-established charge and dipolar asymmetry of the two leaflets of a native membrane, the theory of flexoelectricity (and curvature elasticity) is extended to take into account this asymmetry using linear and nonlinear forms of the Poisson-Boltzmann equation. The results are discussed with respect to data from atomic force microscopy studies of electromotility in biomembranes. PMID- 11863562 TI - Correlated motions of two hydrodynamically coupled particles confined in separate quadratic potential wells. AB - We report a study of the correlated motions of two hydrodynamically coupled colloidal particles, each of which is trapped in a quadratic potential well defined by optical tweezers (optical traps). By setting one of the trapped particles into forced oscillation using oscillating optical tweezers, we measure the displacement and phase shift of each of the particles over a wide frequency range. From the in-phase and out-of-phase motions of both of the particles in the traps, we determine the correlated motions of the coupled mechanical system as a function of frequency. A theoretical model is developed to calculate the response tensor of the coupled mechanical system. The experimental results are in agreement with the prediction of the theoretical model. This method may be extended to more general applications, such as the investigation of the micromechanical properties of viscoelastic and/or heterogeneous media. PMID- 11863563 TI - Reaction-diffusion model for the growth of avascular tumor. AB - A nutrient-limited model for avascular cancer growth including cell proliferation, motility, and death is presented. The model qualitatively reproduces commonly observed morphologies for primary tumors, and the simulated patterns are characterized by its gyration radius, total number of cancer cells, and number of cells on tumor periphery. These very distinct morphological patterns follow Gompertz growth curves, but exhibit different scaling laws for their surfaces. Also, the simulated tumors incorporate a spatial structure composed of a central necrotic core, an inner rim of quiescent cells and a narrow outer shell of proliferating cells in agreement with biological data. Finally, our results indicate that the competition for nutrients among normal and cancer cells may be a determining factor in generating papillary tumor morphology. PMID- 11863564 TI - Predicting the entrainment of reentrant cardiac waves using phase resetting curves. AB - Excitable media, such as the Belousov-Zhabotinsky medium or the heart, are capable of supporting excitation waves that circulate in a closed repetitive path -a phenomenon known as reentrant excitation. A single stimulus, depending on its magnitude, timing, and location, can cause a time shift of the reentrant excitation called resetting. The present study examines the ability of resetting data to predict the effects of periodic stimuli on reentrant excitation circulating on an annular domain. We compare the results of the theoretical models with experiments carried out in an animal model of a dangerous reentrant cardiac rhythm. The current work may lead to improved approaches to therapy through a better understanding of how typical clinical stimuli interact with abnormal reentrant cardiac rhythms. PMID- 11863565 TI - Frequency-domain theory of laser infrared photothermal radiometric detection of thermal waves generated by diffuse-photon-density wave fields in turbid media. AB - A three-dimensional theory of the frequency-domain thermal-wave field generated inside a turbid medium with optical and thermal properties of human tissue is presented. The optical source is treated as a three-dimensional harmonically modulated diffuse-photon-density wave (DPDW) field in the diffusion approximation of the radiative transfer theory. Unlike earlier Green-function-based theoretical models, exact boundary conditions are used based on the requirement that there should be no diffuse photon intensity entering the turbid medium from the outside. Explicit analytical expressions for the DPDW field and for the dependent thermal-wave field are obtained in the spatial Hankel-transform domain. The formalism is further extended to the calculation of the infrared photothermal radiometric signal arising from the nonradiatively generated thermal-wave distribution in turbid media with instantaneous nonradiative deexcitation as well as in media with nonzero fluorescence relaxation lifetimes. Numerical inversions have been performed and presented as examples of selected special cases of the theory. It is found that the present theory with exact DPDW-field boundary conditions is valid throughout the entire domain of the turbid medium, with the exception of the very near-surface ballistic photon "skin layer" (7-50 microm). Photothermal radiometric signals were found to be more reliably predicted than DPDW signals within this layer, due to the depth-integration nature of this detection methodology. PMID- 11863567 TI - Finite-temperature large acoustic polaron dynamics in quasi-one-dimensional molecular crystals. AB - We report the results of theoretical examinations of large polaron motion in one dimensional (1D) molecular crystals under the influence of thermal fluctuations of the host lattice and constant electric field. Such a situation may arise in biological macromolecules such as an alpha helix where charge (electron) transfer may be achieved by a polaron (soliton) mechanism. In that case, the electric field represents the effective endogenous electric field which is always present in realistic conditions. We derive and solve the Fokker-Planck equation for the distribution function of the soliton's center-of-mass position. It is shown that the soliton effectively exhibits a random walk. Moreover, in order to examine statistical properties of the soliton wave function, we calculate the mean value of the soliton probability density: and we find that, for sufficiently large times, thermal fluctuations destruct the soliton, which transforms into the Gaussian packet. These results were used in order to estimate the relevance of the soliton model of charge transfer in polypeptide chains. PMID- 11863566 TI - Equilibrium analysis of the efficiency of an autonomous molecular computer. AB - In the whiplash polymerase chain reaction (WPCR), autonomous molecular computation is implemented in vitro by the recursive, self-directed polymerase extension of a mixture of DNA hairpins. Although computational efficiency is known to be reduced by a tendency for DNAs to self-inhibit by backhybridization, both the magnitude of this effect and its dependence on the reaction conditions have remained open questions. In this paper, the impact of backhybridization on WPCR efficiency is addressed by modeling the recursive extension of each strand as a Markov chain. The extension efficiency per effective polymerase-DNA encounter is then estimated within the framework of a statistical thermodynamic model. Model predictions are shown to provide close agreement with the premature halting of computation reported in a recent in vitro WPCR implementation, a particularly significant result, given that backhybridization had been discounted as the dominant error process. The scaling behavior further indicates completion times to be sufficiently long to render WPCR-based massive parallelism infeasible. A modified architecture, PNA-mediated WPCR (PWPCR) is then proposed in which the occupancy of backhybridized hairpins is reduced by targeted PNA(2)/DNA triplex formation. The efficiency of PWPCR is discussed using a modified form of the model developed for WPCR. Predictions indicate the PWPCR efficiency is sufficient to allow the implementation of autonomous molecular computation on a massive scale. PMID- 11863568 TI - Symmetry scheme for amino acid codons. AB - Group theoretical concepts are invoked in a specific model to explain how only twenty amino acids occur in nature out of a possible sixty four. The methods we use enable us to justify the occurrence of the recently discovered 21st amino acid selenocysteine, and also enables us to predict the possible existence of two more, as yet undiscovered amino acids. PMID- 11863569 TI - Theoretical predictions of electromechanical deformation of cells subjected to high voltages for membrane electroporation. AB - An electromechanical analysis based on thin-shell theory is presented to analyze cell shape changes in response to external electric fields. This approach can be extended to include osmotic-pressure changes. Our calculations demonstrate that at large fields, the spherical cell geometry can be significantly modified, and even ellipsoidal forms would be inappropriate to account for the deformation. Values of the surface forces obtained from our calculations are in very good agreement with the 1--10 mN/m range for membrane rupture reported in the literature. The results, in keeping with reports in the literature, demonstrate that the final shape depends on membrane thickness. This has direct implications for tissues in which significant molecular restructuring can occur. It is also shown that, at least for the smaller electric fields, both the cellular surface area and volume change roughly in a quadratic manner with the electric field. Finally, it is shown that the bending moments are generally quite small and can be neglected for a simpler analysis. PMID- 11863570 TI - Suppression of noise in nonlinear systems subjected to strong periodic driving. AB - An overdamped Brownian motion in "quartic" potential subjected to periodic driving has been considered. This system for the case of a weak periodic driving has been intensively studied during past decade within the context of stochastic resonance. It has been demonstrated that for the case of predominantly suprathreshold driving (the driving amplitude is significantly larger than the static threshold) the noise in the output signal is strongly suppressed at a certain frequency range: the signal-to-noise ratio demonstrates resonant behavior as a function of frequency. PMID- 11863571 TI - Morphology of a solidifying interface near a spherical particle: disjoining pressure effects. AB - Numerical simulations were conducted to determine the morphology of a solid liquid interface near an insoluble spherical particle. The model accounts for the undercoolings due to the front's curvature and to the nonretarded van der Waals interactions. Our numerical results show that, in the near-contact region, the interface profile develops a sharp peak whose curvature has a logarithmic singularity. This is in agreement with the asymptotic analysis of a previously published equation for the interface profile. PMID- 11863572 TI - Segregation of granular binary mixtures by a ratchet mechanism. AB - We report on a segregation scheme for granular binary mixtures, where the segregation is performed by a ratchet mechanism realized by a vertically shaken asymmetric sawtooth-shaped base in a quasi-two-dimensional box. We have studied this system by computer simulations and found that most binary mixtures can be segregated using an appropriately chosen ratchet, even when the particles in the two components have the same size and differ only in their normal restitution coefficient or friction coefficient. These results suggest that the components of otherwise nonsegregating granular mixtures may be separated using our method. PMID- 11863573 TI - Generalized cholesteric permeation flows. AB - The permeation flow equations of cholesteric liquid crystals are derived using a decoupled formulation of the Leslie-Ericksen equations. The formulation sheds light on the role of Ericksen elastic stresses in permeation flows. The Darcy flow regime is shown to emerge in the absence of velocity gradients. The permeation flow equations are generalized to gravity driven flow and used to analyze a free-boundary film flow over an inclined plane. PMID- 11863574 TI - Do effective interactions depend on the choice of coordinates? AB - A common approach to complex systems such as colloidal suspensions or polymer solutions describes the mesoscopic behavior using effective interactions. These potentials act between the macromolecular entities and can be derived by integrating out the microscopic degrees of freedom. The remaining macroparticle coordinates need to be chosen a priori. Two obvious choices are (i) the centers of mass and (ii) distinct microscopic entities, such as special "tagged" monomers. Here we compare both in the framework of the Asakura-Oosawa colloid ideal polymer mixture. Using computer simulations, we find that although the effective pair interaction between colloid and polymer differ markedly, correlation functions are in fair agreement. PMID- 11863575 TI - Wavelet and receiver operating characteristic analysis of heart rate variability. AB - Multiresolution wavelet analysis has been used to study the heart rate variability in two classes of patients with different pathological conditions. The scale dependent measure of Thurner et al. was found to be statistically significant in discriminating patients suffering from hypercardiomyopathy from a control set of normal subjects. We have performed Receiver Operating Characteristc (ROC) analysis and found the ROC area to be a useful measure by which to label the significance of the discrimination, as well as to describe the severity of heart dysfunction. PMID- 11863576 TI - Roughness at the depinning threshold for a long-range elastic string. AB - In this paper, we compute to high precision the roughness exponent zeta of a long range elastic string, at the depinning threshold, in a random medium. Our numerical method exploits the analytic structure of the problem ("no-passing" theorem), but avoids direct simulation of the evolution equations. The roughness exponent has recently been studied by simulations, functional renormalization group calculations, and by experiments (fracture of solids, liquid meniscus in 4He). Our result zeta=0.388 +/- 0.002 is significantly larger than what was stated in previous simulations, which were consistent with a one-loop renormalization-group calculation. Furthermore, the data are incompatible with the experimental results for crack propagation in solids and for a 4He contact line on a rough substrate. This implies that the experiments cannot be described by pure harmonic long-range elasticity in the quasistatic limit. PMID- 11863577 TI - Birhythmicity induced by perturbing an oscillating electrochemical system. AB - We describe the generation of new limit cycles in electrochemical systems under the influence of external periodic perturbations. For certain specific parameters of a nonharmonic forcing function, two coexisting periodic orbits can be generated from a single limit cycle observed in the unperturbed dynamics. This inception of birhythmicity (bistability) is observed in both simulations and actual experiments involving potentiostatic electrodissolution of copper in an acetate buffer. PMID- 11863578 TI - Wave function correlations on the ballistic scale: exploring quantum chaos by quantum disorder. AB - We study the statistics of wave functions in a ballistic chaotic system. The statistical ensemble is generated by adding weak smooth disorder. The conjecture of Gaussian fluctuations of wave functions put forward by Berry [J. Phys. A 10, 2083 (1977)] and generalized by Hortikar and Srednicki [Phys. Rev. Lett. 80, 1646 (1998); Phys. Rev. E 57, 7313 (1998)] is proven to hold on sufficiently short distances, while it is found to be strongly violated on larger scales. This also resolves the conflict between the above conjecture and the wave function normalization. The method is further used to study ballistic correlations of wave functions in a random magnetic field. PMID- 11863579 TI - Localized random lasing modes and a path for observing localization. AB - We demonstrate that a knowledge of the density of states and the eigenstates of a random system without gain, in conjunction with the frequency profile of the gain, can accurately predict the mode that will lase first. Its critical pumping rate can also be obtained. It is found that the shape of the wave function of the random system remains unchanged as gain is introduced. These results were obtained by the time-independent transfer matrix method and finite-difference time-domain methods in a one-dimensional model. They can also be analytically understood by generalizing the semiclassical Lamb theory of lasing in random systems. These findings provide a path for observing the localization of light, such as looking for the mobility edge and studying the localized states. PMID- 11863580 TI - Soliton ratchets. AB - The mechanism underlying the soliton ratchet, both in absence and in presence of noise, is investigated. We show the existence of an asymmetric internal mode on the soliton profile that couples, through the damping in the system, to the soliton translational mode. Effective soliton transport is achieved when the internal mode and the external force are phase locked. We use as a working model a generalized double sine-Gordon equation. The phenomenon is expected to be valid for generic soliton systems. PMID- 11863582 TI - Wealth condensation in pareto macroeconomies. AB - We discuss a Pareto macroeconomy (a) in a closed system with fixed total wealth and (b) in an open system with average mean wealth, and compare our results to a similar analysis in a super-open system (c) with unbounded wealth [J.-P. Bouchaud and M. Mezard, Physica A 282, 536 (2000)]. Wealth condensation takes place in the social phase for closed and open economies, while it occurs in the liberal phase for super-open economies. In the first two cases, the condensation is related to a mechanism known from the balls-in-boxes model, while in the last case, to the nonintegrable tails of the Pareto distribution. For a closed macroeconomy in the social phase, we point to the emergence of a "corruption" phenomenon: a sizeable fraction of the total wealth is always amassed by a single individual. PMID- 11863581 TI - Perturbative method for the derivation of quantum kinetic theory based on closed time-path formalism. AB - Within the closed-time-path formalism, a perturbative method is presented, which reduces the microscopic field theory to the quantum kinetic theory. In order to make this reduction, the expectation value of a physical quantity must be calculated under the condition that the Wigner distribution function is fixed, because it is the independent dynamical variable in the quantum kinetic theory. It is shown that when a nonequilibrium Green function in the form of the generalized Kadanoff-Baym ansatz is utilized, this condition appears as a cancellation of a certain part of contributions in the diagrammatic expression of the expectation value. Together with the quantum kinetic equation, which can be derived in the closed-time-path formalism, this method provides a basis for the kinetic-theoretical description. PMID- 11863583 TI - Crashes, recoveries, and "core shifts" in a model of evolving networks. AB - A model of an evolving network of interacting molecular species is shown to exhibit repeated rounds of crashes in which several species get rapidly depopulated, followed by recoveries. The network inevitably self- organizes into an autocatalytic structure, which consists of an irreducible "core" surrounded by a parasitic "periphery." Crashes typically occur when the existing autocatalytic set becomes fragile and suffers a "core shift," defined graph theoretically. The nature of the recovery after a crash, in particular, the time of recovery, depends upon the organizational structure that survives the crash. The largest eigenvalue of the adjacency matrix of the graph is an important signal of network fragility or robustness. PMID- 11863584 TI - Two-dimensional equilibrium surface roughness for dissociative dimer dynamics. AB - Equilibrium crystal surfaces, constrained to equilibrate by means of dissociative dimer deposition and evaporation, have anomalous global surface roughness. We generalize earlier results for one-dimensional interfaces to two dimensions. The global surface width scales with surface size L as W2 approximately equals ln[L/(ln L)(1/4)] instead of the conventional form W2 approximately equal to ln L. The surface roughening transition does not change in nature, but its location is subject to a large and slowly varying logarithmic finite-size-scaling shift. PMID- 11863585 TI - Tsallis dynamics using the Nose-Hoover approach. AB - On the basis of the Nose-Hoover method, we developed a deterministic algorithm that produces an arbitrary probability density. An ordinary differential equation in the algorithm can realize the Tsallis distribution density. The Tsallis distribution has been considered a candidate of a distribution that represents a physical system in a heat bath. The Tsallis distribution density employed in this algorithm is defined using a full energy function form E(x,p), along with the Tsallis index q > or = 1. Using the current equation, numerical simulations were performed for simple systems and the Tsallis distributions were observed. PMID- 11863586 TI - Traveling kinks in discrete media: exact solution in a piecewise linear model. AB - We present an exact integral representation of a traveling kink solution in a reaction-diffusion equation with a piecewise linear reaction function, complementing existence proofs and numerical observations of such solutions in discrete excitable media. The kink speed is determined through a matching condition, and is worked out explicitly in two limiting situations: the pinning limit, and the opposite limit of infinitely fast kink. Results on the pinning limit agree with those in a recent paper by Fath [Physica D 116, 176 (1998)]. The model includes a "recovery parameter" for a possible extension to a discrete FitzHugh-Nagumo-type system. PMID- 11863587 TI - Growing scale-free networks with tunable clustering. AB - We extend the standard scale-free network model to include a "triad formation step." We analyze the geometric properties of networks generated by this algorithm both analytically and by numerical calculations, and find that our model possesses the same characteristics as the standard scale-free networks such as the power-law degree distribution and the small average geodesic length, but with the high clustering at the same time. In our model, the clustering coefficient is also shown to be tunable simply by changing a control parameter-- the average number of triad formation trials per time step. PMID- 11863588 TI - Surrogate analysis of coherent multichannel data. AB - We present a method for generating surrogate data for multichannel time series. By preserving both the power spectra and the cross spectrum of the original data, we can determine if a given statistical test (such as the synchronization index) is being biased by the presence of coherence due to linear superposition of the separate measurements. Current methods based on phase randomization techniques are unsuitable for this particular task. This method is demonstrated on various canonical systems and numerical models. We will show that this algorithm is capable of properly preserving the power spectra and coherence function of the original data, and furthermore, that with the help of surrogate analysis the synchronization index measure is capable of distinguishing between coupled nonlinear oscillators and coherent superpositions of independent chaotic oscillators. PMID- 11863589 TI - Blume-Capel model approximated by a sequence of generalized Husimi trees. AB - We generalize a systematic approximation method presented by the present author earlier [Monroe, Phys. Rev. E 64, 016126 (2001)], and which was applied to Ising models with spin one-half. The generalization allows one to consider higher spin systems. In particular we consider the spin-one, Blume-Capel model on a square lattice. We obtain an approximation to the phase diagram of the system that we show is as or more accurate than any presently available. This we are able to do with a rather modest effort thereby illustrating the fact that the method gives one rather accurate results without requiring too extensive computer calculations. PMID- 11863590 TI - Direct estimation of the partition function from computer simulation. AB - We propose an approximate method for directly estimating the partition function of classical, many-body model systems. The accessible part of the phase space is determined from a single simulation. We introduce the method for the hard-sphere fluid and solid. The best performance is found in the dense fluid regime, close to freezing density and in the solid crystal. Defining hard-core effective diameters, the method can be applied to systems with soft-core interactions. We present results of exploratory calculations for the Lennard-Jones liquid. PMID- 11863591 TI - Critical behavior of the mixed-spin Ising model with two competing dynamics. AB - In this work we investigate the stationary states of a nonequilibrium mixed-spin Ising model on a square lattice. The model system consists of two interpenetrating sublattices of spins sigma=1/2 and S=1, and we take only nearest neighbor interactions between pairs of spins. The system is in contact with a heat bath at temperature T and subject to an external flux of energy. The contact with the heat bath is simulated by single spin flips according to the Metropolis rule, while the input of energy is mimicked by the simultaneous flipping of pairs of neighboring spins. We performed Monte Carlo simulations on this model in order to find its phase diagram in the plane of temperature T versus the competition parameter between one- and two-spin flips, p. The phase diagram of the model exhibits two ordered phases with sublattice magnetizations m(1), m(2)>0 and m(1)>0, m(2)<0. These phases are separated from the paramagnetic phase (m(1)=m(2)=0) by continuous transition lines. We found the static critical exponents along these lines and showed that this nonequilibrium model belongs to the universality class of the two-dimensional equilibrium Ising model. PMID- 11863592 TI - Spatial prisoner's dilemma games with dynamic payoff matrices. AB - The effects of dynamic payoff matrices on evolution of cooperation are studied based on the prisoner's dilemma game on a two-dimensional square lattice. The study is conducted by simulation and an analytical theory based on mean-field approximation. Payoff matrices are designed to evolve depending on a ratio of defectors (or cooperators) to the whole population. Dynamic payoff matrices are necessary to describe evolution of a society whose payoff may be affected by the results of actions of the members in the society. Introducing such payoff matrices helps to model dynamic aspects of societies. PMID- 11863594 TI - Type of self-organized criticality model based on neural networks. AB - Based on the standard self-organizing map neural network model, we introduce a kind of coupled map lattice system to investigate self-organized criticality (SOC) in the activity of model neural populations. Our system is simulated by a more detailed integrate-and-fire mechanism and a kind of local perturbation driving rule; it can display SOC behavior in a certain range of system parameters, even with period boundary condition. More importantly, when the influence of synaptic plasticity is adequately considered, we can find that our system's learning process plays a promotive role in the emergence of SOC behavior. PMID- 11863595 TI - Solutions for certain number-conserving deterministic cellular automata. AB - We explain the unexpected behavior of the generalizations of cellular automation traffic models introduced in [H. Fuks and N. Boccara, Int. J. Mod. Phys. C 9, 1 (1998)]. We analyze the steady-state flow in R(m,k) as a function of the initial density; we show that these rules correspond to a system with an infinite number of different kinds of virtual particles interacting according to complex annihilation rules. From simple considerations, we are able to predict the unexpected cutoff of the average flow at unity observed by Fuks and Boccara. We present an efficient algorithm for determining the exact final flow from a given finite initial state. An analysis of this algorithm in the infinite limit using generating functions yields an exact polynomial equation between the flow and density for R(m,k), of maximally 2(m+k)th degree in both. PMID- 11863593 TI - Phase diagram and critical exponents of a Potts gauge glass. AB - The two-dimensional q-state Potts model is subjected to a Z(q) symmetric disorder that allows for the existence of a Nishimori line. At q=2, this model coincides with the +/- J random-bond Ising model. For q>2, apart from the usual pure- and zero-temperature fixed points, the ferro/paramagnetic phase boundary is controlled by two critical fixed points: a weak disorder point, whose universality class is that of the ferromagnetic bond-disordered Potts model, and a strong disorder point which generalizes the usual Nishimori point. We numerically study the case q=3, tracing out the phase diagram and precisely determining the critical exponents. The universality class of the Nishimori point is inconsistent with percolation on Potts clusters. PMID- 11863596 TI - Macroscopic dynamics through coarse-graining: a solvable example. AB - The recently derived fluctuation-dissipation formula [A. N. Gorban et al., Phys. Rev. E 63, 066124 (2001)] is illustrated by the explicit computation for McKean's kinetic model [H. P. McKean, J. Math. Phys. 8, 547 (1967)]. It is demonstrated that the result is identical, on the one hand, to the sum of the Chapman-Enskog expansion, and, on the other hand, to the exact solution of the invariance equation. The equality between all three results holds up to the crossover from the hydrodynamic to the kinetic domain. PMID- 11863597 TI - Iterative renormalization group for anomalous dimension in a nonlinear diffusion process. AB - We construct a classical successive method, the Picard method in integral equation theory, to make an iterative algorithm with the renormalization group (RG) approach to calculate the anomalous dimension in a nonlinear diffusion equation. We find our result improves than the original RG work because we begin with the var epsilonth RG solution, not the trivial fixed-point solution. PMID- 11863598 TI - Functional integral approach: a third formulation of quantum statistical mechanics. AB - Quantum statistical mechanics has developed primarily through two approaches, pioneered by Gibbs and Feynman, respectively. In Gibbs' method one calculates partition functions from phase-space integrations or sums over stationary states. Alternatively, in Feynman's approach, the focus is on the path-integral formulation. The Hubbard-Stratonovich transformation leads to a functional integral formulation for calculating partition functions. We outline here the functional integral approach to quantum statistical mechanics, including generalizations and improvements to Hubbard's formulation. We show how the dimensionality of the integrals is reduced exactly, how the problem of assuming an unknown canonical transformation is avoided, how the reality of the partition function in the complex representation is guaranteed, and how the extremum conditions are simplified. This formulation can be applied to general systems, including superconductors. PMID- 11863599 TI - High-frequency measurements of interfacial friction using quartz crystal resonators integrated into a surface forces apparatus. AB - A quartz crystal resonator covered with a thin sheet of mica was integrated into a surface forces apparatus. The shifts in resonance frequency and bandwidth were monitored as the mica surface came into contact with a spherical lens approached from above. We compare experiments where the lens was either coated with a second mica sheet or just had a silver layer evaporated onto its surface. For the contact with the silver surface, strong maxima in bandwidth occurred during the formation and the disruption of the contact. No such maxima were seen when approaching and separating two mica surfaces. We attribute this increased dissipation to sliding and rolling friction, involving plastic deformation of the metal surface under oscillatory load. PMID- 11863600 TI - Simple model for 1/f(alpha) noise. AB - We present a simple stochastic mechanism which generates pulse trains exhibiting a power-law distribution of the pulse intervals and a 1/f(alpha) power spectrum over several decades at low frequencies with alpha close to 1. The essential ingredient of our model is a fluctuating threshold which performs a Brownian motion. Whenever an increasing potential V(t) hits the threshold, V(t) is reset to the origin and a pulse is emitted. We show that if V(t) increases linearly in time, the pulse intervals can be approximated by a random walk with multiplicative noise. Our model agrees with recent experiments in neurobiology and explains the high interpulse interval variability and the occurrence of 1/f(alpha) noise observed in cortical neurons and earthquake data. PMID- 11863601 TI - Multicomponent binary spreading process. AB - I investigate numerically the phase transitions of two-component generalizations of binary spreading processes in one dimension. In these models pair annihilation AA --> emptyset, BB --> emptyset, explicit particle diffusion, and binary pair production processes compete with each other. Several versions with spatially different production are explored, and it is shown that for the cases 2A --> 3A, 2B--> 3B and 2A --> 2AB, 2B--> 2BA a phase transition occurs at zero production rate (sigma=0), which belongs to the class of N-component, asymmetric branching and annihilating random walks, characterized by the order parameter exponent beta=2. In the model with particle production AB --> ABA, BA --> BAB a phase transition point can be located at sigma(c)=0.3253 which belongs to the class of one-component binary spreading processes. PMID- 11863602 TI - Speed selection mechanism for propagating fronts in reaction-diffusion systems with multiple fields. AB - We introduce a speed selection mechanism for front propagation in reaction diffusion systems with multiple fields. This mechanism applies to pulled and pushed fronts alike, and operates by restricting the fields to large finite intervals in the comoving frames of reference. The unique velocity for which the center of a monotonic solution for a particular field is insensitive to the location of the ends of the finite interval is the velocity that is physically selected for that field, making thus the solution approximately translation invariant. The fronts for the various fields may propagate at different speeds, all of them being determined though through this mechanism. We present analytic results for the case of piecewise parabolic potentials, and numerical results for other cases. PMID- 11863603 TI - Metastable structures with modified weighted density-functional theory. AB - The free energy of the supercooled liquid near freezing is studied in the density functional approach using the modified weighted density approximation. A class of minima corresponding to heterogeneous structures characterized by weak mass localization are detected. The stability of these structures is found to be greater than the highly localized "hard-sphere glass" state in the intermediate density range above freezing. PMID- 11863604 TI - Exact relations between critical exponents for elastic stiffness and electrical conductivity of two-dimensional percolating networks. AB - It has long been known that the critical exponent T of the elastic stiffness C(e)proportional, Deltap(T) of a d-dimensional percolating network (Deltap identical with p - p(c)>0 measures the closeness of the network to its percolation threshold p(c)) satisfies the following inequalities: 1+dnu < or = T < or = t+2nu, where t is the critical exponent of the electrical conductivity sigma(e) proportional, Deltap(t) of the same network and nu is the critical exponent of the percolation correlation length xi proportional, Deltap(-nu). Similarly, the critical exponents that characterize the divergences C(e)proportional, /Deltap/(-S), sigma(e) proportional to /Deltap/(-s) of a rigid or normal and a superconducting or normal random mixture (Deltap identical with p (c)<0 now measures the closeness of the rigid or superconducting constituent to its percolation threshold p(c)) have long been known to satisfy S < or = s. We now show that, when d=2, T is in fact exactly equal to t+2nu and S is exactly equal to s. PMID- 11863605 TI - Conjugate gradient filtering of instantaneous normal modes, saddles on the energy landscape, and diffusion in liquids. AB - Instantaneous normal modes (INM's) are calculated during a conjugate-gradient (CG) descent of the potential energy landscape, starting from an equilibrium configuration of a liquid or crystal. A small number (approximately equal to 4) of CG steps removes all the Im-omega modes in the crystal and leaves the liquid with diffusive Im-omega which accurately represent the self-diffusion constant D. Conjugate gradient filtering appears to be a promising method, applicable to any system, of obtaining diffusive modes and facilitating INM theory of D. The relation of the CG-step dependent INM quantities to the landscape and its saddles is discussed. PMID- 11863606 TI - Boundary polarization in the six-vertex model. AB - Vertical-arrow fluctuations near the boundaries in the six-vertex model on the two-dimensional NxN square lattice with the domain wall boundary conditions are considered. The one-point correlation function ("boundary polarization") is expressed via the partition function of the model on a sublattice. The partition function is represented in terms of standard objects in the theory of orthogonal polynomials. This representation is used to study the large N limit: the presence of the boundary affects the macroscopic quantities of the model even in this limit. The logarithmic terms obtained are compared with predictions from conformal field theory. PMID- 11863607 TI - Stretched exponential relaxation in a diffusive lattice model. AB - We studied the single dimer dynamics in a lattice diffusive model as a function of particle density in the high densification regime. The mean square displacement is found to be subdiffusive both in one and two dimensions. The spatial dependence of the self-part of the van Hove correlation function displays as a function of r a single peak and signals a dramatic slow down of the system for high density. The self-intermediate scattering function is fitted to the Kohlrausch-Williams-Watts law. The exponent beta extracted from the fits is density independent while the relaxation time tau follows a scaling law with an exponent 2.5. PMID- 11863608 TI - Statistical mechanics of relativistic one-dimensional self-gravitating systems. AB - We consider the statistical mechanics of a general relativistic one-dimensional self-gravitating system. The system consists of N particles coupled to lineal gravity and can be considered as a model of N relativistically interacting sheets of uniform mass. The partition function and one-particle distribution functions are computed to leading order in 1/c where c is the speed of light; as c --> infinity results for the nonrelativistic one-dimensional self-gravitating system are recovered. We find that relativistic effects generally cause both position and momentum distribution functions to become more sharply peaked, and that the temperature of a relativistic gas is smaller than its nonrelativistic counterpart at the same fixed energy. We consider the large-N limit of our results and compare this to the nonrelativistic case. PMID- 11863609 TI - Finite-size scaling in disordered systems. AB - The critical behavior of a quenched random hypercubic sample of linear size L is considered, within the "random-T(c)" field-theoretical model, by using the renormalization group method. A finite-size scaling behavior is established and analyzed near the upper critical dimension d=4-epsilon and some universal results are obtained. The problem of self-averaging is clarified for different critical regimes. PMID- 11863610 TI - Group symmetries in two-body random matrix ensembles generating order out of complexity. AB - The two-body random matrix ensembles with spin TBRE-s and in a single j shell TBRE-j introduced recently in the context of ground state structures in complex interacting particle systems, possess U(N) superset U(N/2)multiply sign in circle SU(2) and U(N)superset O(3) group symmetries, respectively, with N the number of single particle states. It is shown that both these group symmetries give rise to simplicities in the ground state structures but in different ways. PMID- 11863611 TI - Dynamics of spinodal decomposition in finite-lifetime systems: Nonlinear statistical theory based on a coarse-grained lattice-gas model. AB - We study theoretically dynamics of the spinodal decomposition in finite-lifetime systems to clarify effects of the interparticle interactions beyond the Ginzburg Landau-Wilson phenomenology. Our theory is based on the coarse-grained Hamiltonian derived from the interacting lattice-gas model with a finite lifetime. The information of a system is reduced to closed-form coupled integrodifferential equations for the single-point distribution function and the dynamical structure factor. These equations involve explicitly the interparticle interactions. The finite lifetime prevents the phase separation and the order formation in the cw creation case; domains cannot grow to be larger than an asymptotic characteristic size [k(max)(t --> infinity)](-1). Power-law dependence of k(max)(t --> infinity) on the interparticle interaction and the particle lifetime is also found numerically. The finite lifetime prevents the phase separation, i.e., the lower critical wave number k((1))(c) appears and domains of size larger than [k((1))(c)](-1) cannot grow. PMID- 11863612 TI - Statistical theory for the Kardar-Parisi-Zhang equation in (1+1) dimensions. AB - The Kardar-Parisi-Zhang (KPZ) equation in (1+1) dimensions dynamically develops sharply connected valley structures within which the height derivative is not continuous. We develop a statistical theory for the KPZ equation in (1+1) dimensions driven with a random forcing that is white in time and Gaussian correlated in space. A master equation is derived for the joint probability density function of height difference and height gradient P(h-h*, partial differential(x)h,t) when the forcing correlation length is much smaller than the system size and much larger than the typical sharp valley width. In the time scales before the creation of the sharp valleys, we find the exact generating function of h-h* and partial differential(x)h. The time scale of the sharp valley formation is expressed in terms of the force characteristics. In the stationary state, when the sharp valleys are fully developed, finite-size corrections to the scaling laws of the structure functions left angle bracket(h-h*)(n)(partial differential(x)h)(m)right angle bracket are also obtained. PMID- 11863613 TI - Formation of vortex loops (strings) in continuous phase transitions. AB - The formation of vortex loops (global cosmic strings) in an O(2) linear sigma model in three spatial dimensions is analyzed numerically. For over-damped Langevin dynamics we find that defect production is suppressed by an interaction between correlated domains that reduces the effective spatial variation of the phase of the order field. The degree of suppression is sensitive to the quench rate. A detailed description of the numerical methods used to analyze the model is also reported. PMID- 11863614 TI - Scale-invariant behavior in a spatial game of prisoners' dilemma. AB - A spatially extended version of the game of prisoner's dilemma, originally proposed by Nowak and May, is modified to include stochastic updating and found to exhibit scale-invariant behavior. Two critical regimes with different scaling behaviors are found; the corresponding exponents have been determined numerically. Spatially, the critical states are characterized by the existence of delicately balanced networks of defectors separating domains of cooperators; temporally, the evolution of the critical states following local perturbations is characterized by avalanches of various magnitudes, which cause restructuring of the networks of defectors on all scales. PMID- 11863615 TI - Self-organized critical forest-fire model on large scales. AB - We discuss the scaling behavior of the self-organized critical forest-fire model on large length scales. As indicated in earlier publications, the forest-fire model does not show conventional critical scaling, but has two qualitatively different types of fires that superimpose to give the effective exponents typically measured in simulations. We show that this explains not only why the exponent characterizing the fire-size distribution changes with increasing correlation length, but allows us also to predict its asymptotic value. We support our arguments by computer simulations of a coarse-grained model, by scaling arguments and by analyzing states that are created artificially by superimposing the two types of fires. PMID- 11863616 TI - Width distributions and the upper critical dimension of Kardar-Parisi-Zhang interfaces. AB - Simulations of restricted solid-on-solid growth models are used to build the width distributions of d=2-5 dimensional Kardar-Parisi-Zhang (KPZ) interfaces. We find that the universal scaling function associated with the steady-state width distribution changes smoothly as d is increased, thus strongly suggesting that d=4 is not an upper critical dimension for the KPZ equation. The dimensional trends observed in the scaling functions indicate that the upper critical dimension is at infinity. PMID- 11863617 TI - Occam factors and model independent Bayesian learning of continuous distributions. AB - Learning of a smooth but nonparametric probability density can be regularized using methods of quantum field theory. We implement a field theoretic prior numerically, test its efficacy, and show that the data and the phase space factors arising from the integration over the model space determine the free parameter of the theory ("smoothness scale") self-consistently. This persists even for distributions that are atypical in the prior and is a step towards a model independent theory for learning continuous distributions. Finally, we point out that a wrong parametrization of a model family may sometimes be advantageous for small data sets. PMID- 11863618 TI - Anomalous diffusion in the nonasymptotic regime. AB - We analyze some properties of the one-dimensional Levy flights, assuming that the one-step transition rates depend on the flight length x as p(alpha)(x) equivalent to x(-(alpha+2)). For flights on a finite, (2M+1)-site lattice, we can define an effective, size-dependent, diffusion coefficient D(alpha)(M) equivalent to [M(1 alpha) - 1]/(1 - alpha) if alpha < 1, with D1(M) equivalent to ln(M). Using the generalization of statistical mechanics given by Tsallis, we show that for flights on infinite systems, the generalized displacement moments are well defined provided that alpha > R - 3. These moments exhibit a power-law singularity if alpha --> 1(-) and R>2/3. The short- and intermediate-time properties of the generalized mean-square displacement are then studied numerically. This work suggests the conditions under which the asymptotic analytical formulas (obtained in the literature by the use of the generalized central limit theorem) could be applied to finite-time experiments. These formulas should work much better if alpha is close to zero than in the alpha - >1(-) neighborhood. PMID- 11863619 TI - Synchronization on small-world networks. AB - We investigate collective synchronization in a system of coupled oscillators on small-world networks. The order parameters that measure synchronization of phases and frequencies are introduced and analyzed by means of dynamic simulations and finite-size scaling. Phase synchronization is observed to emerge in the presence of even a tiny fraction P of shortcuts and to display saturated behavior for P > or similar to 0.5. This indicates that the same synchronizability as the random network (P=1) can be achieved with relatively small number of shortcuts. The transient behavior of the synchronization, obtained from the measurement of the relaxation time, is also discussed. PMID- 11863620 TI - High temperature behavior of a two-parameter deformed quantum group fermion gas. AB - We discuss a two-parameter deformed quantum group fermion gas with SU(q/p)(2) symmetry. In order to obtain the role of the deformation parameters (q,p) on the thermodynamics of the system, we calculate several thermodynamical functions and investigate the high temperature behavior of a SU(q/p)(2) fermion gas through a SU(q/p)(2)-invariant fermionic Hamiltonian. However, the ordinary fermion gas results can be obtained by applying the limit q=p=1. PMID- 11863621 TI - Dynamics of phase separation in multicomponent mixtures. AB - We study the dynamics of phase separation in multicomponent mixtures through Monte Carlo simulations of the q-state Potts model with conserved kinetics. We use the Monte Carlo renormalization-group method to investigate the asymptotic regime. The domain growth law is found to be consistent with the Lifshitz-Slyozov law, L(t) equivalent to t(1/3) (where t is time), regardless of the value of q. We also present results for the scaled correlation functions and domain-size distribution functions for a range of q values. PMID- 11863622 TI - Factorized time correlation diagram analysis of paired causal systems excited by twin stochastic driving functions. AB - This work examines the properties and mathematical structure of factorized time correlation (FTC) diagram analysis in a general context. The goal is to extract general principles and analytic behavior that are not tied to any particular phenomenon in physics. It is hoped that this will provide a basis for expanded use of FTC diagram analysis beyond its current employment in the study of noisy light-based nonlinear optical spectroscopy. Furthermore, the concept of indirect correlation in a two-channel system driven by twin stationary circular Gaussian stochastic inputs is defined and discussed both analytically and through FTC diagram analysis. PMID- 11863623 TI - Quantum dissipation in unbounded systems. AB - In recent years trajectory based methodologies have become increasingly popular for evaluating the time evolution of quantum systems. A revival of the de Broglie -Bohm interpretation of quantum mechanics has spawned several such techniques for examining quantum dynamics from a hydrodynamic perspective. Using techniques similar to those found in computational fluid dynamics one can construct the wave function of a quantum system at any time from the trajectories of a discrete ensemble of hydrodynamic fluid elements (Bohm particles) which evolve according to nonclassical equations of motion. Until very recently these schemes have been limited to conservative systems. In this paper, we present our methodology for including the effects of a thermal environment into the hydrodynamic formulation of quantum dynamics. We derive hydrodynamic equations of motion from the Caldeira Leggett master equation for the reduced density matrix and give a brief overview of our computational scheme that incorporates an adaptive Lagrangian mesh. Our applications focus upon the dissipative dynamics of open unbounded quantum systems. Using both the Wigner phase space representation and the linear entropy, we probe the breakdown of the Markov approximation of the bath dynamics at low temperatures. We suggest a criteria for rationalizing the validity of the Markov approximation in open unbound systems and discuss decoherence, energy relaxation, and quantum/classical correspondence in the context of the Bohmian paths. PMID- 11863624 TI - Walker diffusion method for calculation of transport properties of finite composite systems. AB - A heterogeneous medium may be represented by a scalar field of local transport coefficients (e.g., conductivity) or by a "resistor network" derived from that scalar field. In either case the effective (macroscopic) and local (microscopic) transport properties may be calculated by the walker diffusion method. Some sample calculations for disordered systems are presented to demonstrate the method. PMID- 11863625 TI - Interface depinning versus absorbing-state phase transitions. AB - According to recent numerical results from lattice models, the critical exponents of systems with many absorbing states and order parameter coupled to a nondiffusive conserved field coincide with those of the linear interface depinning model within computational accuracy. In this paper the connection between absorbing-state phase transitions and interface pinning in quenched disordered media is investigated. For that, we present an heuristic mapping of the interface dynamics in a disordered medium into a Langevin equation for the active-site density and show that a Reggeon-field-theory-like description, in which the order parameter appears coupled to an additional nondiffusive conserved field, emerges rather naturally. Reciprocally, we construct a mapping from a discrete model belonging in the absorbing state with a conserved-field class to a discrete interface equation, and show how a quenched disorder, typical of the interface representation is originated. We discuss the character of the possible noise terms in both representations, and overview the critical exponent relations. Evidence is provided that, at least for dimensions larger that one, both universality classes are just two different representations of the same underlying physics. PMID- 11863627 TI - Hints for universality in coupled map lattices. AB - A wide range of coupled map lattices is found to have identical pattern sequences providing numerical evidence for their universality in the class of systems studied. It is furthermore found that the wave-type solutions of the diffusively coupled logistic lattice scale linearly with the coupling range indicating the existence of a continuum limit. The findings are used to introduce a very simple type of spatially extended map that can serve as a representative for the pattern sequence of this class of coupled maps. PMID- 11863626 TI - Two regimes of synchronization in unidirectionally coupled semiconductor lasers. AB - We analyze unidirectionally coupled semiconductor lasers in the feedback/injection scheme to determine their synchronization performance. As the mismatch between the two lasers increases, there is a transition from complete synchronization for identical lasers to time lag synchronization which is only partial. This corresponds to a continuous change of the global minimum that becomes a relative minimum of the synchronization error function and vice versa. PMID- 11863628 TI - Sequential fragmentation: the origin of columnar quasihexagonal patterns. AB - We present a model that explains the origin and predicts the statistical properties of columnar quasihexagonal crack patterns, as observed in the columnar jointing of basaltic lava flows. Induced by temperature gradients during cooling, irregular fractures appear at the surface of the material. At later times fractures penetrate into the material, and tend to form polygonal patterns. We show that this ordering can be described as a tendency to minimize an energy functional. Atomistic simulations confirm this interpretation. Numerical simulations based on a phenomenological implementation of this principle generate patterns that have remarkably good statistical agreement with real ones. PMID- 11863630 TI - Choice of dynamical variables for global reconstruction of model equations from time series. AB - The success of modeling from an experimental time series is determined to a significant extent by the choice of dynamical variables. We propose a method for preliminary investigation of a time series whose purpose is to find out whether a global dynamical model with smooth functions can be constructed for the chosen variables. The method consists in the estimation of single valuedness and continuity of relations between dynamical variables and variables to enter left hand sides of model equations. The method is explained with numerical examples. Its efficiency is demonstrated by modeling a real nonlinear electric circuit. PMID- 11863629 TI - External feedback control of chaos using approximate periodic orbits. AB - We apply the external feedback technique to control chaos in real physical systems. The target, unstable periodic orbits embedded in chaotic attractors are obtained from chaotic time series in terms of the delay coordinates technique. We demonstrate its efficiency for periodically forced, single- and two-degree-of freedom systems consisting of one or two pendula in numerical simulations and experiments. PMID- 11863631 TI - Instabilities of the resonance attractor for spiral waves in an excitable medium. AB - During an experimental study of the resonance attractor for spiral waves in the light-sensitive Belousov-Zhabotinsky reaction, strong deviations of the attractor trajectories from circular orbits are observed if the time delay in the feedback loop becomes relatively long. A theory is developed that reduces the spiral wave dynamics under a long-delayed control to a higher order iterative map. Then the observed deviations are explained to be a result of instabilities appearing due to the Neimark bifurcation of the map. The theoretical predictions are in good agreement with the experimental data. PMID- 11863632 TI - Controlling chaos in unidimensional maps using macroevolutionary algorithms. AB - We introduce a simple search algorithm that explores the parameter of periodically perturbed discrete maps in order to find desired orbits through chaos control. The method has been applied to one-dimensional maps but is easily extendable to higher-dimensional systems. Here, we consider two types of chaos control involving proportional pulses in the system variables [Phys. Rev. Lett. 72, 1455 (1994)] and constant feedback [Phys. Rev. E 51, 6239 (1995)], the first case being presented in detail. It is shown that our method allows a rapid exploration of parameter space and the finding of high-fitness (i.e., controlled) solutions close to the target orbits, even when high periodicities are required. PMID- 11863633 TI - Spontaneous phase oscillation induced by inertia and time delay. AB - We consider a system of coupled oscillators with finite inertia and time-delayed interaction, and investigate the interplay between inertia and delay both analytically and numerically. The phase velocity of the system is examined; revealed in numerical simulations is the emergence of spontaneous phase oscillation without external driving, which turns out to be in good agreement with analytical results derived in the strong-coupling limit. Such self oscillation is found to suppress synchronization, and its frequency is observed to decrease with inertia and delay. We obtain the phase diagram, which displays oscillatory and stationary phases in the appropriate regions of the parameters. PMID- 11863634 TI - Unexpected robustness against noise of a class of nonhyperbolic chaotic attractors. AB - Chaotic attractors arising in physical systems are often nonhyperbolic. We compare two sources of nonhyperbolicity: (1) tangencies between stable and unstable manifolds, and (2) unstable dimension variability. We study the effects of noise on chaotic attractors with these nonhyperbolic behaviors by investigating the scaling laws for the Hausdorff distance between the noisy and the deterministic attractors. Whereas in the presence of tangencies, interactive noise yields attractor deformations, attractors with only dimension variability are robust, despite the fact that shadowing is grossly violated. PMID- 11863636 TI - Stochastic ionization through noble tori: renormalization results. AB - We find that chaos in the stochastic ionization problem develops through the breakup of a sequence of noble tori. In addition to being very accurate, our method of choice, the renormalization map, is ideally suited for analyzing properties at criticality. Our computations of chaos thresholds agree closely with the widely used empirical Chirikov criterion. PMID- 11863635 TI - Characterization of the parameter-mismatching effect on the loss of chaos synchronization. AB - We investigate the effect of the parameter mismatch on the loss of chaos synchronization in coupled one-dimensional maps. Loss of strong synchronization begins with a first transverse bifurcation of a periodic saddle embedded in the synchronous chaotic attractor (SCA), and then the SCA becomes weakly stable. Because of local transverse repulsion of the periodic repellers embedded in the weakly stable SCA, a typical trajectory may have segments of arbitrary length that have positive local transverse Lyapunov exponents. Consequently, the weakly stable SCA becomes sensitive with respect to the variation of the mismatching parameter. To quantitatively characterize such parameter sensitivity, we introduce a quantifier, called the parameter sensitivity exponent (PSE). As the local transverse repulsion of the periodic repellers strengthens, the value of the PSE increases. In terms of these PSEs, we also characterize the parameter mismatching effect on the intermittent bursting and basin riddling occurring in the regime of weak synchronization. PMID- 11863637 TI - Self-induced slow-fast dynamics and swept bifurcation diagrams in weakly desynchronized systems. AB - In systems close to the state of phase synchronization, the fast timescale of oscillations interacts with the slow timescale of the phase drift. As a result, "fast" dynamics is subjected to a slow modulation, due to which an autonomous system under fixed parameter values can imitate repeated bifurcational transitions. We demonstrate the action of this general mechanism for a set of two coupled autonomous chaotic oscillators and for a chaotic system perturbed by a periodic external force. In both cases, the Poincare sections of phase portraits resemble bifurcation diagram of a logistic mapping with time-dependent parameter. PMID- 11863638 TI - Universality of chaotic rare fluctuations in a locally coupled phase map model. AB - Chaotic fluctuations of the order parameter in a coupled two-dimensional phase map model are numerically investigated. We discuss the system-size N dependence of the statistical properties of rare fluctuations observed in the transition range between the quasiordered chaotic state and the fully developed one. It is found that the normalized probability distribution function (PDF) has a unique functional form irrespective of N. The asymptotic form of the PDF is discussed in connection with the universal distribution for correlated systems proposed by Bramwell et al. [Nature (London) 396, 552 (1998)]. Moreover, it is observed that the power spectrum P(N)(omega) of rare fluctuations asymptotically takes the power-law form P(N)(omega) equivalent to omega(-(1+alpha)) (alpha=0.6 equivalent to 0.7) irrespective of N. This result suggests that the temporal correlation decays as a stretched exponential. PMID- 11863639 TI - Signatures of the correlation hole in total and partial cross sections. AB - In a complex scattering system with few open channels, say a quantum dot with leads, the correlation properties of the poles of the scattering matrix are most directly related to the internal dynamics of the system. We may ask how to extract these properties from an analysis of cross sections. In general this is very difficult, if we leave the domain of isolated resonances. We propose to consider the cross correlation function of two different elastic or total cross sections. For these we can show numerically and to some extent also analytically a significant dependence on the correlations between the scattering poles. The difference between uncorrelated and strongly correlated poles is clearly visible, even for strongly overlapping resonances. PMID- 11863640 TI - Driving trajectories in chaotic scattering. AB - In this work we introduce a general approach for targeting in chaotic scattering that can be used to find a transfer trajectory between any two points located inside the scattering region. We show that this method can be used in association with a control of chaos strategy to drive around and keep a particle inside the scattering region. As an illustration of how powerful this approach is, we use it in a case of practical interest in celestial mechanics in which it is desired to control the evolution of two satellites that evolve around a large central body. PMID- 11863641 TI - Finite-size effects on active chaotic advection. AB - A small (but finite-size) spherical particle advected by fluid flows obeys equations of motion that are inherently dissipative, due to the Stokes drag. The dynamics of the advected particle can be chaotic even with a flow field that is simply time periodic. Similar to the case of ideal tracers, whose dynamics is Hamiltonian, chemical or biological activity involving such particles can be analyzed using the theory of chaotic dynamics. Using the example of an autocatalytic reaction, A+Bright arrow2B, we show that the balance between dissipation in the particle dynamics and production due to reaction leads to a steady state distribution of the reagent. We also show that, in the case of coalescence reaction, B+Bright arrowB, the decay of the particle density obeys a universal scaling law as approximately t(minus sign1) and that the particle distribution becomes restricted to a subset with fractal dimension D2, where D2 is the correlation dimension of the chaotic attractor in the particle dynamics. PMID- 11863642 TI - Branch points in the complex plane and geometric phases. AB - Laser-induced degenerate states (LIDS) are equivalent to double poles of the S matrix that are branch points in the complex plane (BPCP). These branch points cause geometric phase changes by encircling them adiabatically around a closed circuit by varying certain parameters. They cause also the well-known phase changes appearing by encircling a diabolic point (DP) being a singularity associated with level repulsion. In both cases, the wave functions are exchanged, Phi(i) --> +/- iPhi(j not equal i), at the critical value of the parameter where the states avoid crossing. Such a critical point is passed twice by encircling a DP but only once by surrounding a BPCP. As a consequence, the phase changes are different in both cases. A second surrounding restores the wave functions including their phases in both cases (when the BPCP is well isolated from others and the time of encircling is shorter than the lifetime of the two states). The different interference pictures appearing in surrounding LIDS adiabatically in opposite directions on a closed circuit represent a completion of the work by Berry. PMID- 11863643 TI - Dephasing due to the interaction with chaotic degrees of freedom. AB - We consider the motion of a particle, taking into account its interaction with environmental degrees of freedom. The dephasing time is determined by the nature of the environment and depends on the particle velocity. Our interest is in the case where the environment consists of few chaotic degrees of freedom. We obtain results for the dephasing time and compare them with those of the effective-bath approach. The latter approach is based on the conjecture that the environment can be modeled as a collection of infinitely many harmonic oscillators. The work is related to studies of driven systems, quantum irreversibility, and fidelity. The specific model that we consider requires the solution of the problem of a particle in a box with a moving wall, whose one-dimensional version is related to the Fermi acceleration problem. PMID- 11863644 TI - Control of chaotic solitons by a time-delayed feedback mechanism. AB - We investigate a control of chaotic solitons using a modified time-delayed autosynchronization method. Numerical results for the maximal Lyapunov exponent are in very good agreement with analytical theory developed originally for low dimensional systems. The control is most efficient when the spatial distribution of the control force is proportional to the kink translational mode. Observations of the motion of the kink center and the total power transmitted to the system lead to the same values of the maximal Lyapunov exponent. PMID- 11863645 TI - Fractal spin glass properties of low energy configurations in the Frenkel Kontorova chain. AB - We study, numerically and analytically, the classical one-dimensional Frenkel Kontorova chain in the regime of pinned phase characterized by phonon gap. Our results show the existence of exponentially many static equilibrium configurations that are exponentially close to the energy of the ground state. The energies of these configurations form a fractal quasidegenerate band structure that is described on the basis of elementary excitations. Contrary to the ground state, the configurations inside these bands are disordered. PMID- 11863646 TI - Density of proper delay times in chaotic and integrable quantum billiards. AB - We calculate the density P(tau) of the eigenvalues of the Wigner-Smith time delay matrix for two-dimensional rectangular and circular billiards with one opening. For long times, the density of these so-called "proper delay times" decays algebraically, in contradistinction to chaotic quantum billiards for which P(tau) exhibits a long-time cutoff. PMID- 11863647 TI - Mean electromotive force in turbulent shear flow. AB - We consider the mean electromotive force in turbulent shear flow taking into account the stretching of turbulent magnetic field lines by the mean flow. The mean flow can change the properties of magnetohydrodynamics-turbulence in such a way that turbulent motions become suitable for the dynamo action. The contribution of shear to the mean electromotive force cannot be described in terms of the alpha effect. The instability of the mean field arises if shear is sufficiently strong. The growth rate of instability depends on the length scale of the mean field being higher for the field with a smaller length scale. The considered mechanism may be responsible for the generation of large-scale magnetic fields in various astrophysical bodies (galaxies, accretion discs, jets, etc.). PMID- 11863648 TI - Statistical mechanics of the shallow water system. AB - We extend the formalism of statistical mechanics developed for the two dimensional (2D) Euler equation to the case of shallow water system. Relaxation equations towards the maximum entropy state are proposed, which provide a parametrization of subgrid-scale eddies in 2D compressible turbulence. PMID- 11863649 TI - Analytical approach to viscous fingering in a cylindrical Hele-Shaw cell. AB - We report analytical results for the development of the viscous fingering instability in a cylindrical Hele-Shaw cell of radius a and thickness b. We derive a generalized version of Darcy's law in such cylindrical background, and find it recovers the usual Darcy's law for flow in flat, rectangular cells, with corrections of higher order in b/a. We focus our interest on the influence of the cell's radius of curvature on the instability characteristics. Linear and slightly nonlinear flow regimes are studied through a mode-coupling analysis. Our analytical results reveal that linear growth rates and finger competition are inhibited for an increasingly larger radius of curvature. The absence of tip splitting events in cylindrical cells is also discussed. PMID- 11863651 TI - Vibration-induced interfacial instabilities in viscoelastic fluids. AB - Vertically vibrated interfaces between viscoelastic fluids may arise in contexts as diverse as ultrasonic emulsification, microgravity materials processing, and geophysics. If the vibration amplitude is large enough at a given frequency, the interface can become unstable and give rise to standing waves. The present work provides a linear analysis of this phenomenon for the cases where either or both of the fluids are viscoelastic. The fluids are assumed to be laterally unbound, and Floquet theory is used to develop a recursion relation between the temporal modes of the interfacial deformation. Conversion of this relation into a matrix eigenvalue problem allows determination of the critical vibration amplitude needed to excite the standing waves and the corresponding critical wave number. Using a single-mode Maxwell model to describe the viscoelasticity and considering infinite fluid depths, we present calculations for three cases: bottom fluid viscoelastic/top fluid Newtonian, bottom fluid Newtonian/top fluid viscoelastic, and both fluids viscoelastic. When only one of the fluids is viscoelastic, the interfacial waves can respond harmonically to the forcing. The waves may also be excited more easily than in the case where both fluids are Newtonian. When both of the fluids are viscoelastic, it appears possible to excite Stoneley-like waves at the interface. PMID- 11863650 TI - Predator-prey encounters in turbulent waters. AB - With reference to studies of predator-prey encounters in turbulent waters, we demonstrate the feasibility of an experimental method for investigations of particle fluxes to an absorbing surface in turbulent flows. A laboratory experiment is carried out, where an approximately homogeneous and isotropic turbulent flow is generated by two moving grids. The simultaneous trajectories of many small neutrally buoyant polystyrene particles are followed in time. Selecting one of these to represent a predator, while the others are considered as prey, we obtain estimates for the time variation of the statistical average of the prey flux into a suitably defined "sphere of interception." The variation of this flux with the radius in the sphere of interception, as well as the variation with basic flow parameters is well described by a simple model, in particular for radii smaller than a characteristic length scale for the turbulence. Also the Eulerian counterpart of the problem has been analyzed, and the particle fluxes from the two studies compared. PMID- 11863652 TI - Stability of connected cylindrical liquid bridges. AB - Two cylindrical liquid bridges, with a conduit to facilitate flow of liquid from one bridge to the other, were levitated against gravity in a magnetic field gradient. The stability limit of the bridges subjected to near zero total body force was measured as a function of their slenderness ratios, and found to be in good agreement with theoretical predictions. PMID- 11863654 TI - Effects of periodicity on flow and dispersion through closely packed fixed beds of spheres. AB - A lattice-Boltzmann formulation is used to investigate the effects of "periodicity" (geometry) on fluid flow and tracer-particle dispersion through fixed beds of spheres comprising of closely packed layers. In the "period-1" arrangement, spheres in the adjacent layers contact at their poles while the "period-2" and "period-3" arrangements correspond to hexagonal and faced-centered cubic close packing. For all three packing arrangements, there is a transition with increasing Reynolds number from a power law to a log-normal distribution of kinetic energies and, velocity and vorticity become more closely aligned giving rise to helical tracer-particle trajectories. It is suggested that these flow characteristics, unlike the stability of flow and the distribution of helicity, are largely insensitive to geometry, even when the geometry creates direct channels through the pack bed orientated along the gradient in applied pressure. For steady flows and strongly turbulent flows, such channels are predicted to provide direct routes for dispersion through a packed bed, while for weakly turbulent flows they influence dispersion primarily by destabilizing the flow and thereby promoting dispersion throughout a bed. The dispersion of tracer-particles released from a source located on or close to a "stagnation streamline" is predicted to be faster than ballistic in the near field and the transition to long-time Fickian diffusion is predicted to be distinguished by a regime of subdiffusion. PMID- 11863653 TI - Statistics of velocity gradients in two-dimensional Navier-Stokes and ocean turbulence. AB - Probability density functions and conditional averages of velocity gradients derived from upper ocean observations are compared with results from forced simulations of the two-dimensional Navier-Stokes equations. Ocean data are derived from TOPEX satellite altimeter measurements. The simulations use rapid forcing on large scales, characteristic of surface winds. The probability distributions of transverse velocity derivatives from the ocean observations agree with the forced simulations, although they differ from unforced simulations reported elsewhere. The distribution and cross correlation of velocity derivatives provide clear evidence that large coherent eddies play only a minor role in generating the observed statistics. PMID- 11863655 TI - Quasisolitons and asymptotic multiscaling in shell models of turbulence. AB - A variation principle is suggested to find self-similar solitary solutions (referred to as solitons) of shell model of turbulence. For the Sabra shell model the shape of the solitons is approximated by rational trial functions with relative accuracy of O(10(-3)). It is found how the soliton shape, propagation time t(n) (from a shell n to shells with n --> infinity), and the dynamical exponent z(0) (which governs the time rescaling of the solitons in different shells) depend on parameters of the model. For a finite interval of z the author discovered quasisolitons which approximate with high accuracy corresponding self similar equations for an interval of times from -infinity to some time in the vicinity of the peak maximum or even after it. The conjecture is that the trajectories in the vicinity of the quasisolitons (with continuous spectra of z) provide an essential contribution to the multiscaling statistics of high-order correlation functions, referred to in the paper as an asymptotic multiscaling. This contribution may be even more important than that of the trajectories in the vicinity of the exact soliton with a fixed value z(0). Moreover there are no solitons in some regions of the parameters where quasisolitons provide a dominant contribution to the asymptotic multiscaling. PMID- 11863656 TI - One-point statistics of the induced electric field in quasinormal magnetofluid turbulence. AB - We study one-point statistical properties of the induced turbulent electric field for a magnetohydrodynamic (MHD) plasma under the quasinormal approximation. Assuming exact Gaussianity for both the velocity field and the magnetic field, and different degrees of correlations between their Cartesian components, we derive the probability distribution function (PDF) for the Cartesian components of the electric field e(i). We show that the PDF reduces in some canonical cases to an exponential function of the form exp(-/e(i)/). To study deviations from these results in the more realistic case in which the velocity and magnetic fields are not exactly normal but quasinormal instead, we perform three dimensional numerical simulations of the MHD equations at moderate Reynolds numbers. For turbulent relaxation from an initial condition, we find that the analytical results give a very good first-order approximation to the computed PDF. PMID- 11863657 TI - Quasiperiodic waves at the onset of zero-Prandtl-number convection with rotation. AB - We show the possibility of temporally quasiperiodic waves at the onset of thermal convection in a thin horizontal layer of slowly rotating zero-Prandtl-number Boussinesq fluid confined between stress-free conducting boundaries. Two independent frequencies emerge due to an interaction between straight rolls and waves along these rolls in the presence of Coriolis force, if the Taylor number is raised above a critical value. Constructing a dynamical system for the hydrodynamical problem, the competition between the interacting instabilities is analyzed. The forward bifurcation from the conductive state is self-tuned. PMID- 11863658 TI - Instability of a thin film flowing on a rotating horizontal or inclined plane. AB - In this paper the instability of a thin fluid film flowing under the effects of gravity, Coriolis, and centrifugal forces is investigated. It is supposed that the film flows far from the axis of rotation on a plane which may be horizontal or inclined with respect to the horizontal. In the former case, the flow is only driven by the centrifugal force while in the latter case, the flow is driven by the components of centrifugal force and gravity along the plane. This case may also be considered as the flow down a rotating cone but far from the apex. The stabilizing influence of rotation on the film flow increases with the rotation rate. Up to a certain critical rate of rotation, the film flowing down the rotating inclined plane (or cone) is more stable than the flow on the horizontal rotating plane while above this rate of rotation the situation is reversed. The instability above the critical rate is associated with a finite wave number in contrast to the vanishing wave number of the instability below the critical rate. The possibility of Ekman layer instabilities is also investigated. An equation describing the nonlinear evolution of surface waves is also obtained. Moreover, this equation is simplified for the case in which the amplitudes are very small. An equation including dissipation as well as dispersion is derived whose solutions may possess solitary waves, as in the case of similar equations considered in the literature. These solutions are likely to correspond to the solitary spiral waves observed in experiments. PMID- 11863659 TI - Shear instability of fluid interfaces: stability analysis. AB - We examine the linear stability of fluid interfaces subjected to a shear flow. Our main object is to generalize previous work to an arbitrary Atwood number, and to allow for surface tension and weak compressibility. The motivation derives from instances in astrophysical systems where mixing across material interfaces driven by shear flows may significantly affect the dynamical evolution of these systems. PMID- 11863660 TI - Statistically preserved structures in shell models of passive scalar advection. AB - It was conjectured recently that statiscally preserved structures underlie the statistical physics of turbulent transport processes. We analyze here in detail the time-dependent (noncompact) linear operator that governs the dynamics of correlation functions in the case of shell models of passive scalar advection. The problem is generic in the sense that the driving velocity field is neither Gaussian nor delta correlated in time. We show how to naturally discuss the dynamics in terms of an effective compact operator that displays "zero modes," which determine the anomalous scaling of the correlation functions. Since shell models have neither a Lagrangian structure nor "shape dynamics," this example differs significantly from standard passive scalar advection. Nevertheless, with the necessary modifications, the generality and efficacy of the concept of statistically preserved structures are further exemplified. In passing we point out a bonus of the present approach, in providing analytic predictions for the time-dependent correlation functions in decaying turbulent transport. PMID- 11863661 TI - Poiseuille-type flow of a rarefied gas between two parallel plates driven by a uniform external force. AB - A unidirectional flow of a rarefied gas between two parallel plates driven by a uniform external force is investigated on the basis of kinetic theory with special interest in the behavior in the near continuum regime. The Bhatnagar Gross-Krook (BGK) model of the Boltzmann equation and the diffuse reflection boundary condition are employed as the basic system. First, a systematic asymptotic analysis of the basic system for small Knudsen numbers is carried out, and a system of fluid-dynamic-type equations and their boundary conditions are derived up to the second order in the Knudsen number. Then, an accurate numerical analysis of the original BGK system is performed for a wide range of the Knudsen number by means of a finite-difference method. The behavior of the gas, such as the non-Navier-Stokes effects in the near continuum regime, is clarified on the basis of the fluid-dynamic-type system as well as the numerical solution of the BGK system. PMID- 11863662 TI - Determination of the line emission locations in a large helical device on the basis of the Zeeman effect. AB - Neutral helium He I lambda 728.1 nm (2 1P-3 1S) and lambda 667.8 nm (2 1P-3 1D) emission lines have been observed with an array of optical fibers that cover the entire poloidal cross section of the plasma. The Zeeman profile yields a magnetic field strength, and the locations of the emission regions are identified on the well-established map of the magnetic field of the plasma. It is found that the emission region forms a closed zone just outside the region, the so-called "ergodic layer," where the magnetic field line structure is chaotic. A collisional-radiative model calculation for an inward atom flux suggests a peaked emission profile of about 3.5 cm in full width at half maximum, and this is consistent with the experimental result. The inward atom flux is found to decay before reaching the last closed flux surface and this implies a screening effect of the ergodic layer. PMID- 11863663 TI - Statistical mechanics of axisymmetric vortex rings. AB - We construct maximum entropy states of a collection of interacting uniform (omega/R=const) axisymmetric vortex rings in a semiperiodic bounded volume. Following Miller [Phys. Rev. Lett. 65, 2137 (1990)] and Robert and Sommeria [J. Fluid Mech. 229, 291 (1991)], we obtain an equilibrium measure that preserves all the ideal invariants such as the total energy, total impulse, circulation, and an infinity of Casimirs. The numerical solution for a wide range of total flow energy and for given values of total circulation and total impulse is presented. PMID- 11863664 TI - Enhancement of x-ray line emission from plasmas produced by short high-intensity laser double pulses. AB - Femtosecond laser-produced plasmas are bright ultrafast line x-ray sources potentially suitable for different applications including material science and biology. The conversion efficiency of the laser energy incident onto a solid target into the x-ray emission is significantly enhanced when a laser prepulse precedes the main pulse. The details of x-ray line emission from solid targets irradiated by a pair of ultrashort laser pulses are investigated both theoretically and experimentally. Insight into spatial and temporal characteristics of the line x-ray source is provided by numerical simulations and a simplified analytical model. Optimal time separation of the laser pulses is searched for in order to reach the maximum conversion of laser energy into the emission of selected x-ray lines. We deduced how the optimal pulse separation scales with laser and target parameters. PMID- 11863665 TI - High-power-density capillary discharge plasma columns for shorter wavelength discharge-pumped soft-x-ray lasers. AB - We report the generation of plasma columns in gas-filled capillary channels using discharge excitation powers that exceed those of previous studies by one to two orders of magnitude. Current pulses up to 200 kA and 10-90 % rise time of about 10 ns (current increase rate equivalent to 1.5 x 10(13) A/s) were utilized to excite plasmas in 3.3 and 4 mm diameter channels. Time resolved soft-x-ray spectra and pinhole images of the plasma were obtained. The experimental data and its comparison with model computations suggest that dense argon plasma columns 300 mum in diameter with electron temperatures >250 eV have been obtained. These characteristics make these plasmas of interest for extending discharge-pumped lasers to shorter wavelengths. PMID- 11863666 TI - Eigenmodes for capillary tubes with dielectric walls and ultraintense laser pulse guiding. AB - The properties of the eigenmodes of a capillary tube are examined in the context of ultrashort intense laser pulse guiding. The dispersion relation for the eigenmodes of a cylindrical hollow waveguide is derived and the family of eigenmodes EH(nus) is shown to be a solution of the wave equation up to the first order under the condition k(0)a >>1, where k(0) is the light wave number and a the capillary tube radius. The expressions of the fields for the eigenmodes are given at zero and first order of a small parameter equal to the ratio of the perpendicular to longitudinal wave number and the absorbed intensity at the wall is estimated. PMID- 11863667 TI - Coherent structures in a turbulent environment. AB - A systematic method is proposed for the determination of the statistical properties of a field consisting of a coherent structure interacting with turbulent linear waves. The explicit expression of the generating functional of the correlations is obtained, performing the functional integration on a neighborhood in the function space around the soliton. The results show that the non-Gaussian fluctuations observed in the plasma edge can be explained by the intermittent formation of nonlinear coherent structures. PMID- 11863668 TI - How wrong is collisional Monte Carlo modeling of fast electron transport in high intensity laser-solid interactions? AB - The interaction of a high-intensity laser with a solid target generates a large current of fast electrons flowing into the target. Due to the large value of the current, the fast electrons generate significant electric and magnetic fields in the target and rapidly heat it to high temperatures. However, these effects were neglected in interpreting x-ray emission experiments, so the details of the fast electron generation that were inferred could be incorrect. This is considered, theoretically, for layered target, Kalpha emission experiments, by using a hybrid Monte Carlo code that includes field generation. The code is used to model such experiments with aluminum and plastic targets, using fast electron parameters taken from experimental results for average intensities of around 10(18) W cm( 2). These numerical results are then interpreted in the same manner as previous experiments, using only the Monte Carlo part of the code. The field generation leads to lower total emission and to an apparent two-temperature fast electron distribution. The laser absorption into fast electrons inferred by Monte Carlo modeling is consistently lower than the actual value. The mean fast electron energy inferred could be either higher or lower than the actual value, depending on the experimental setup and the cone angle and energy distribution used in the Monte Carlo modeling. The errors caused by neglecting the fields are, in general, greater for plastic than aluminum targets, leading to inconsistencies in results obtained by Monte Carlo modeling. PMID- 11863669 TI - Generalized theory and simulation of spontaneous and super-radiant emissions in electron devices and free-electron lasers. AB - A unified formulation of spontaneous (shot-noise) and super-radiant emissions in electron devices is presented. We consider an electron beam with an arbitrary temporal current modulation propagating through the interaction region of the electronic device. The total electromagnetic field is presented as a stochastic process and expanded in terms of transverse eigenmodes of the medium (free space or waveguide), in which the field is excited and propagates. Using the waveguide excitation equations, formulated in the frequency domain, an analytical expression for the power spectral density of the electromagnetic radiation is derived. The spectrum of the excited radiation is shown to be composed of two terms, which are the spontaneous and super-radiant emissions. For a continuous, unmodulated beam, the shot noise produces only incoherent spontaneous emission of a power proportional to the flux eI(0) (DC current) of the particles in the electron beam. When the beam is modulated or prebunched, a partially coherent super-radiant emission is also produced with power proportional to the current spectrum /I(omega)/(2). Based on a three-dimensional model, a numerical particle simulation code was developed. A set of coupled-mode excitation equations in the frequency domain are solved self-consistently with the equations of particles motion. The simulation considers random distributions of density and energy in the electron beam and takes into account the statistical and spectral features of the excited radiation. At present, the code can simulate free-electron lasers (FELs) operation in various modes: spontaneous and self-amplified spontaneous emission, super-radiance and stimulated emission, in the linear and nonlinear Compton or Raman regimes. We employed the code to demonstrate spontaneous and super-radiant emission excited when a prebunched electron beam passes through a wiggler of an FEL. PMID- 11863670 TI - Heating rate of hadron beams during crystallization. AB - A theory concerning the relation between the heating rate and temperature of hadron beams is formulated from a quantum point of view. This theory predicts that the heating rate can be reduced by increasing the lattice periodicity of the accelerator with its fixed tunes and circumference. This prediction is quite consistent with simulation results. PMID- 11863671 TI - Observation of matter wave beat phenomena in the macrodomain for electrons moving along a magnetic field. AB - We report here the observations that exhibit the existence of matter wave phenomena with wavelength in the macrodomain of a few centimeters, for electrons moving along a magnetic field from an electron gun to a collector plate situated behind a grounded grid. These are in accordance with the predictions of a quantumlike theory for charged particles in the classical macrodomain, given by one of the authors [R. K. Varma, Phys. Rev. A 31, 3951 (1985)] with a recent generalization [R. K. Varma, Phys. Rev. E 64, 036608 (2001)]. The beats correspond to two closely spaced "frequencies" in the system, with the beat frequency given, in accordance with the characteristics of a wave phenomena, by the difference between the two frequencies. The beats ride as a modulation over a discrete energy band structure obtained with only one frequency present. The frequency here corresponds to the distance between the electron gun and the detector plate as it characterizes the variation in the energy band structure as the electron energy is swept. The second "frequency" corresponds to the gun-grid distance. These observations of the beats of matter waves in this experiment, with characteristics in accordance with the wave algorithm, then establish unambiguously the existence of macroscopic matter waves for electrons propagating along a magnetic field. PMID- 11863672 TI - Escape angles in bulk chi((2)) soliton interactions. AB - We develop a theory for nonplanar interaction between two identical type I spatial solitons propagating at opposite, but arbitrary transverse angles in quadratic nonlinear (or so-called chi((2))) bulk media. We predict quantitatively the outwards escape angle, below which the solitons turn around and collide, and above which they continue to move-away from each other. For in-plane interaction, the theory allows prediction of the outcome of a collision through the inwards escape angle, i.e., whether the solitons fuse or cross. We find an analytical expression determining the inwards escape angle using Gaussian approximations for the solitons. The theory is verified numerically. PMID- 11863673 TI - Methods for discrete solitons in nonlinear lattices. AB - A method to find discrete solitons in nonlinear lattices is introduced. Using nonlinear optical waveguide arrays as a prototype application, both stationary and traveling-wave solitons are investigated. In the limit of small wave velocity, a fully discrete perturbative analysis yields formulas for the mode shapes and velocity. PMID- 11863674 TI - Trapping, reflection, and fragmentation in a classical model of atom-lattice collisions. AB - A classical one-dimensional model of the collision of an atom of mass M with a cold, semi-infinite harmonic lattice comprised of identical atoms of mass m is considered. In the model, the interactions between the incident atom (adatom) and the lattice are described in terms of a truncated parabolic potential by which the adatom is harmonically bound to the lattice at short distances but evolves freely when its distance is larger than a critical length R(c). The dynamics of the adatom colliding with an infinitely cold lattice is studied as a function of the initial velocity of the adatom. In order to determine whether the colliding atom is bound or reflected from the lattice in the asymptotic time limit, "secondary" collision events in which the incident atom leaves and reenters the interaction zone of the lattice are carefully considered. It is demonstrated that secondary collisions anticipated to be important for heavy adatoms (mu=m/M<1) also occur in the case of light adatoms (mu > or = 1). It is shown that the neglect of secondary collisions leads to an underestimation of the lower energy bound for adatom reflection of roughly 10% for mu close to 1. By generalizing the model to allow for the breaking of lattice bonds, the phenomenon of collision induced lattice fragmentation is investigated. PMID- 11863675 TI - Stability of one-dimensional array solitons. AB - The array soliton stability in the discrete nonlinear Schrodinger equation with dispersion for periodic boundary conditions is studied. The linear growth rate dependence on the discrete wave number and soliton amplitude is calculated from the linearized eigenvalue problem using the variational method. In addition, the eigenvalue problem is solved numerically by shooting method and a good agreement with the analytical results is found. It is proved numerically that the results for the instability threshold for the circular array coincides with the quasicollapse threshold for the case of open arrays with initial pulses in a form of array solitons. PMID- 11863676 TI - Interaction of a kink soliton with a breather in a Fermi-Pasta-Ulam chain. AB - The collision process between a breather and moving kink soliton is investigated both analytically and numerically in Fermi-Pasta-Ulam (FPU) chains. As it is shown by both analytical and numerical consideration low amplitude breathers and soft kinks retain their shapes after interaction. Low amplitude breather only changes the location after collision and remains static. As the numerical simulations show, the shift of its position is proportional to the stiffness of the kink soliton, what is in accordance with the analytical predictions made in this paper. The numerical experiments are also carried out for large amplitude breathers and some interesting effects are observed: The odd parity large amplitude breather does not change position when colliding with a widely separated soft kink-antikink pair, while in the case of a closely placed kink antikink pair the breather transforms into the moving one. Therefore it is suggested that the "harmless" objects similar to the kink solitons in FPU chains could be used in order to displace or move the strongly localized structures in realistic physical systems. In particular, the analogies with quasi-one dimensional easy-plane-type spin structures are discussed. PMID- 11863677 TI - Diffraction effects in few-cycle optical pulses. AB - Basic concepts of three-dimensional wave packets are applied to the description of transverse effects on the propagation of ultrashort (femtosecond) pulses. The frequency-dependent nature of diffraction acts as a kind of dispersion that modifies the pulse front surface, its group velocity, the envelope form, and the carrier frequency. If the diffracted field in the monochromatic case is known, these changes can be straightforwardly quantified. Finding the propagated pulsed beam field reduces to a well-known and simpler problem of one-dimensional pulse propagation with group velocity dispersion. The method is applied to pulsed Gaussian beams and pulsed Bessel beams. Anomalous pulse front behavior, including superluminality in pulsed Gaussian beams is found. The carrier phase at any point of space is calculated. PMID- 11863678 TI - Second-harmonic generation with pulses in a coupled-resonator optical waveguide. AB - We describe the generation and propagation of pulses in a coupled-resonator optical waveguide driven by a nonlinear polarization using a method closely related to the coupled-mode theory. The specific example we consider is that of second-harmonic generation. This formalism explicitly accounts for temporal dependencies in the waveguide field distributions and in their representations in terms of slowly modulated Bloch wave functions, in contrast with the equations obtained previously for cw second-harmonic generation. PMID- 11863679 TI - Fluorescence depolarization in a scattering medium: effect of size parameter of a scatterer. AB - For a monodisperse scattering medium, we investigate the dependence on scatterer size parameter for the change in anisotropy of fluorescence due to single scattering at excitation or emission wavelength. The value for the ratio of the anisotropy of fluorescence after one scattering at excitation or emission wavelength to the initial value was observed to increase with increasing value of scatterer size parameter. The effect of multiple scattering on anisotropy of fluorescence from fluorophores embedded in a scattering medium was incorporated using a photon migration model. The model was validated by experiments carried out on samples with known concentration of polystyrene microspheres as scatterers and riboflavins or reduced form of nicotinamide adenine dinucleotide as fluorophores. PMID- 11863680 TI - Solitary waves and supersonic reaction front in metastable solids. AB - Motivated by an increasing number of remarkable experimental observations on the role of pressure and shear stress in solid reactions, explosions, and detonations, we present a simple one-dimensional model that embodies nonlinear elasticity and dispersion as well as chemical or phase transformation. This generalization of the Toda lattice provides an effective model for the description of the organization during an abrupt transformation in a solid. One of the challenges is to capture both the equilibrium degrees of freedom as well as to quantify the possible role of out-of-equilibrium perturbations. In the Toda lattice, we verify that the particle velocities converge in distribution towards the Maxwell-Boltzmann distribution, thus allowing us to define a bonafide temperature. In addition, the balance between nonlinearity and wave dispersion may create solitary waves that act as energy traps. In the presence of reactive chemistry, we show that the trapping of the released chemical energy in solitary waves that are excited by an initial perturbation provides a positive feedback that enhances the reaction rate and leads to supersonic explosion front propagation. These modes of rupture observed in our model may provide a first order description of ultrafast reactions of heterogeneous mixtures under mechanical loading. PMID- 11863681 TI - Spectral properties of fluctuating electromagnetic fields in a plane cavity: implication for nanoscale physics. AB - Spectral power densities of fluctuating electromagnetic fields and their spatial derivatives of all orders in any point of a transparent plane gap between two media described by different complex permittivities and by different temperatures were derived on a basis of generalized Kirchhoff's law. Electromagnetic losses into the two absorbing media induced by a field of a point dipole or of point multipolelike origins situated in any place of interest at the transparent gap were determined. The corresponding electrodynamical regular Green problem for a point dipole and for point multipoles of any orders constituted by the point dipole was solved. We demonstrate ways to obtain different asymptotic cases following from our general solution including the problem for a half space, Planck's formula for black body radiation, the van der Waals forces for solids kept at different temperatures, and contributions from propagating and evanescent waves. Expressions for electromagnetic loss of a point multipole of any order in selected geometry of the problem were derived and, as an important limiting case related to problems of near field microscopy, when the multipole is situated over a half space. PMID- 11863682 TI - Ground states and vortices of matter-wave condensates and optical guided waves. AB - We analyze the shape and stability of localized states in nonlinear cubic media with space-dependent potentials modeling an inhomogeneity. By means of a static variational approach, we describe the ground states and vortexlike stationary solutions, either in dilute atom gases or in optical cavities, with an emphasis on parabolic-type potentials. First, we determine the existence conditions for soliton and vortex structures for both focusing and defocusing nonlinearity. It is shown that, even for a defocusing medium, soliton modes can exist with a confining potential. Second, step potentials and boundedness effects in hollow capillaries are investigated, which both proceed from a similar analysis. Finally, we discuss applications of this procedure to charged vortices in dilute quantum gases and to Bose-Einstein condensates trapped in the presence of a light induced Gaussian barrier. PMID- 11863683 TI - Effective geometry for light traveling in material media. AB - Working with electrodynamics in the geometrical optics approximation, we derive the expression representing the "effective geometry" seen by electromagnetic waves propagating in media whose physical properties depend on an external electric field. Some previous results are generalized and some special cases are recovered. PMID- 11863684 TI - Periodic solutions for systems of coupled nonlinear Schrodinger equations with five and six components. AB - Systems of coupled nonlinear Schrodinger (CNLS) equations arise in several branches of physics, e.g., optics and plasma physics. Systems with two or three components have been studied intensively. Recently periodic solutions for CNLS systems with four components are derived. The present work extends the search of periodic solutions for CNLS systems to those with five and six components. The Hirota bilinear method, theta and elliptic functions are employed in the process. The long wave limit is studied, and known results of solitary waves are recovered. The validity of these periodic solutions is verified independently by direct differentiation with computer algebra software. PMID- 11863685 TI - Correlations among angular wave component amplitudes in elastic multiple scattering random media. AB - The propagation of scalar waves through random media that provide multiple elastic scattering is considered by derivation of an expression for the angular correlation of the scattered wave amplitudes. Coherent wave transmission is shown to occur through a mechanism similar to that responsible for coherent backscattering. While the properties of the scattered wave are generally consistent with radiative-transfer theory for sufficiently small incident and scattering angles, coherent transmission provides corrections to radiative transfer results at larger angles. The theoretical angular correlation curves are fit, by specifying the probability densities of two random variables that correspond to material parameters, to measured data of laser light scattering from various polymer microsphere suspensions. PMID- 11863687 TI - Excitability of excitons and biexcitons in a ring cavity. AB - We discuss the excitable behavior of excitons and biexcitons in a nonlinear optical ring cavity. The nonlinearity is due to the process of the creation of biexcitons by photon-assisted conversion of excitons. In the bifurcation analysis a region where a saddle point is close to an equilibrium has been found. In this region the system shows excitability. It is shown that the mechanism of the excitable behavior of excitons and biexcitons in a ring cavity is different from that of two-level atoms in the same system. The possible applications of an excitable ring cavity are discussed. PMID- 11863686 TI - Discrete Burridge-Knopoff model, with exact solitonic or compactlike traveling wave solution. AB - We have explored the dynamics of two versions of a Burridge-Knopoff model: with linear or nonlinear interactions between adjacent blocks. We have shown that by properly choosing the analytical form of the discrete solitary wave solution of the model we can calculate analytically the form of the friction function. In both cases our analytical results show that the friction force naturally presents the behavior of a simple weakening friction law first introduced qualitatively by Burridge and Knopoff [Bull. Seismol. Soc. Am. 57, 3411 (1967)] and quantitatively by Carlson and Langer [Phys. Rev. Lett. 62, 2632 (1989)]. With such a force function the discrete solitonic or compactlike wave-front solutions are exact and stable solutions. In the case of linear coupling our numerical simulations show that an irregular initial state evolves into kink pairs (large-amplitude events), that can recombine or not, plus nonlinear localized modes and small linear oscillations (small-amplitude events) that disperse with time, owing to dispersion. For nonlinear coupling one observes compactlike kink pairs or shocks, and a background of robust incoherent nonlinear oscillations (small amplitude events) that persist with time. Our results show that discreteness is a necessary ingredient to observe a rich and complex dynamical behavior. Nonlinearity allows the existence of strictly localized shocks. PMID- 11863689 TI - Invaded cluster simulations of the XY model in two and three dimensions. AB - The invaded cluster algorithm is used to study the XY model in two and three dimensions up to sizes 2000(2) and 120(3), respectively. A soft spin O(2) model, in the same universality class as the three-dimensional XY model, is also studied. The static critical properties of the model and the dynamical properties of the algorithm are reported. The results are K(c)=0.454 12(2) for the three dimensional XY model and eta=0.037(2) for the three-dimensional XY universality class. For the two-dimensional XY model the results are K(c)=1.120(1) and eta=0.251(5). The invaded cluster algorithm does not show any critical slowing for the magnetization or critical temperature estimator for the two-dimensional or three-dimensional XY models. PMID- 11863688 TI - The unexplained accuracy of the Lagrange-mesh method. AB - The Lagrange-mesh method is an approximate variational calculation which resembles a mesh calculation because of the use of a Gauss quadrature. In order to analyze its accuracy, four different Lagrange-mesh calculations based on the zeros of Laguerre polynomials are compared with exact variational calculations based on the corresponding Laguerre basis. The comparison is performed for three solvable radial potentials: the Morse, harmonic-oscillator, and Coulomb potentials. The results show that the accuracies of the energies obtained for different partial waves with the different mesh approximations are very close to the variational accuracy, even in the presence of the centrifugal singularity. The same property holds for the approximate wave functions. This striking accuracy remains unexplained. PMID- 11863691 TI - Purification of correlated reduced density matrices. AB - The notion of purification is generalized to treat correlated reduced density matrices. Traditionally, purification denotes the process by which a one-particle reduced density matrix (1-RDM) is made idempotent, that is, its eigenvalues are mapped to either 0 or 1. Purification of correlated RDMs is defined as the iterative process by which an arbitrary RDM is forced to satisfy several necessary N-representability conditions. Using the unitary decomposition of RDMs and the positivity conditions, we develop an algorithm to purify the 2-RDM. The algorithm is applied within the solution of the contracted Schrodinger equation CSE for the 2-RDM [D. A. Mazziotti, Phys. Rev. A 57, 4219 (1998)]. Previous attempts to solve the CSE by powerlike methods have frequently produced divergent energies, but we show that the purification process eliminates the divergent behavior for systematic and reliable convergence of the contracted power method to the N-particle energy. PMID- 11863692 TI - Dimensional crossover and driven interfaces in disordered ferromagnets. AB - We study the depinning transition of driven interfaces in thin ferromagnetic films driven by external magnetic fields. Approaching the transition point the correlation length increases with decreasing driving. If the correlation length becomes of the order of the film thickness a crossover to two-dimensional behavior occurs. From the corresponding scaling analysis, we determine the exponents characterizing the transition of the three-dimensional system. PMID- 11863690 TI - Phase space deformation and basis set optimization. AB - By deforming a given region of phase space---occupied by some unknown eigenfunctions one wishes to find---into a standard, integrable region, effective reductions in basis set size can be achieved. In one-dimensional problems we are able to achieve B/C=1+O(Planck' s constant), where B is the basis set size and C is the number of "converged" eigenfunctions. This result is confirmed by numerical examples, which also indicate exponential convergence as the basis set size is increased. In higher dimensions we prove that such an optimistic result is impossible; we expect that the best one can do in this case is B/C=a+o(1), where a>1 has a geometric interpretation in terms of ratios of volumes in phase space. PMID- 11863693 TI - Exactly solved dynamics for an infinite-range spin system. II. Antiferromagnetic interactions. AB - In a previous paper [E. Milotti, Phys. Rev. E 63, 026116 (2001)]. I have shown how to derive both thermodynamical and dynamical properties of an infinite-range Ising spin system with binary ferromagnetic interactions from the master equation for magnetization obtained from a simple spin dynamics. The same method can be adapted to different spin interactions: here I discuss the case of antiferromagnetic interactions. This model permits a study of the static properties of the antiferromagnetic lattice, and it displays very clearly the differences between the antiferromagnetic and the ferromagnetic case with long range interactions. The dynamical behavior of the antiferromagnetic system is simpler, and the magnetization always relaxes exponentially. PMID- 11863694 TI - Path finding strategies in scale-free networks. AB - We numerically investigate the scale-free network model of Barabasi and Albert [A. L. Barabasi and R. Albert, Science 286, 509 (1999)] through the use of various path finding strategies. In real networks, global network information is not accessible to each vertex, and the actual path connecting two vertices can sometimes be much longer than the shortest one. A generalized diameter depending on the actual path finding strategy is introduced, and a simple strategy, which utilizes only local information on the connectivity, is suggested and shown to yield small-world behavior: the diameter D of the network increases logarithmically with the network size N, the same as is found with global strategy. If paths are sought at random, D is equivalent to N(0.5) is found. PMID- 11863695 TI - Universal persistence exponents in an extremally driven system. AB - The local persistence R(t), defined as the proportion of the system still in its initial state at time t, is measured for the Bak-Sneppen model. For one and two dimensions, it is found that the decay of R(t) depends on one of two classes of initial configuration. For a subcritical initial state, R(t) equivalent to t( theta), where the persistence exponent theta can be expressed in terms of a known universal exponent. Hence theta is universal. Conversely, starting from a supercritical state, R(t) decays by the anomalous form 1-R(t) equivalent to t(tau(all)) until a finite time t(0), where tau(all) is also a known exponent. Finally, for the high dimensional model R(t) decays exponentially with a nonuniversal decay constant. PMID- 11863696 TI - Parameter evaluation from time sequences using chaos synchronization. AB - Unknown parameters in nonlinear equations are estimated from chaotic time sequences using chaos synchronization. The method is based on a random optimization method. The parameters are randomly searched for in a sequential manner as the degree of the chaos synchronization is increased. The method is applied for the parameter evaluation in the Lorenz equation and the Lang Kobayashi model for the chaotic semiconductor laser. PMID- 11863697 TI - Active synchronization in nonhyperbolic hyperchaotic systems. AB - We propose a methodology to address the outstanding problem of synchronization in nonhyperbolic hyperchaotic physical systems. Our approach makes use of a controlling-chaos strategy that accomplishes the task by transmitting only one scalar signal even in the presence of noise. PMID- 11863698 TI - Markov process built in scale-similar multifractal energy cascades in turbulence. AB - The scale-similar multifractal cascade, which is believed to describe the process of energy cascade in the inertial scale-range of isotropic turbulence, is proved to be Markovian so as to be governed by the Chapman-Kolmogorov equation, when negative logarithmic length scale is taken for time. For a limited class of cascades, the Kramers-Moyal expansion of the Chapman-Kolmogorov equation is possible and the coefficients are exactly derived from the functional form of intermittency exponents mu(q), and they are all constant if the cascade is scale similar. PMID- 11863699 TI - Wavelet analysis of vortex tubes in experimental turbulence. AB - This paper proposes a method to study vortex tubes in one-dimensional velocity data of experimental turbulence. Vortex tubes are detected as local maxima on the scale-space plot of wavelet transforms of the velocity data. Then it is possible to extract a typical velocity pattern. The result for data obtained in a wind tunnel is consistent with those of three-dimensional direct numerical simulations. PMID- 11863700 TI - Modulational instability of short-wavelength ion waves in strongly coupled dusty plasmas. AB - Modulational instability of electrostatic short-wavelength ion waves in plasmas containing strongly coupled dusts with variable charge is considered. The evolution equations for the ion waves modulated by slow dust motion are obtained. The instability behavior differs considerably from that of plasmas with weakly coupled dust grains and depends strongly on the Coulomb coupling parameter as well as the dust-charge relaxation rate. PMID- 11863701 TI - Stability of periodic paraxial optical systems. AB - Based on ray propagation of paraxial geometric optics, we show that any stable periodic paraxial system or optical resonator becomes unstable in presence of stochastic perturbations of the the periodic sequence along which the rays are propagated. The exponential divergence with distance of ray displacements from the optical axis bears a close connection to the phenomenon of Anderson localization in disordered systems. The stability of the periodic focusing system is restored when finite aperture effects are accounted for and complex paraxial optics is used to describe wave propagation. PMID- 11863702 TI - Comment on "Vacuum electron acceleration by coherent dipole radiation". AB - Troha et al. [Phys. Rev. E 60, 926 (1999)] put forward a generalized Lawson Woodward theorem in the study of laser accelerations. We point out that one of the assumptions used in their proof does not stand on a solid physical ground and that it is possible for electrons to obtain net energy gains from a plane-wave laser pulse in vacuum even if the radiation reaction effects are neglected. PMID- 11863704 TI - Comment on "Spectral filters in quantum mechanics: a measurement theory perspective". AB - We criticize a paper by Vijay and Wyatt [Phys. Rev. E 63, 4351 (2000)], in which the authors suggest that energy levels computed, from the same set of matrix vector products, with the filter diagonalization method (FDM) and the Fourier spectral analysis using the same Chebyshev correlation function are of comparable accuracy. We explain why the FDM is superior and demonstrate it numerically, using the same test matrix as that employed in the above paper. We also compare the FDM with the Lanczos method, another commonly used iterative technique for computing eigenvalues. We find that eigenvalues in a low-density region near the middle of the spectrum converge more quickly with the FDM, but that the Lanczos method requires fewer matrix-vector products to converge all the eigenvalues. PMID- 11863705 TI - Reply to Comment on "Spectral filters in quantum mechanics: A measurement theory perspective". AB - We address the issues raised in the preceding comment by Mandelshtam and Carrington [Phys. Rev. E 65, 028701 (2002)], concerning the eigenvalue determination by the spectral filter methods. We argue that the Fourier transformation is the building block of all currently known time-domain spectral filter algorithms, and, therefore, the time-energy uncertainty principle affects them all in a similar manner. We also explain the situation when the correlation function method may be less suitable, in comparison to the filter diagonalization method, for the determination of eigenvalues, even though both share the same fundamental principles. PMID- 11863706 TI - Generic entanglement generation, quantum statistics, and complementarity. AB - A general and an arbitrarily efficient scheme for entangling the spins (or any spinlike degree of freedom) of two independent uncorrelated identical particles by a combination of two particle interferometry and which way detection is formulated. It is shown that the same setup could be used to identify the quantum statistics of the incident particles from either the sign or the magnitude of measured spin correlations. Our setup also exhibits a curious complementarity between particle distinguishability and the amount of generated entanglement. PMID- 11863707 TI - Conservation laws, uncertainty relations, and quantum limits of measurements. AB - The uncertainty relation between the noise operator and the conserved quantity leads to a bound on the accuracy of general measurements. The bound extends the assertion by Wigner, Araki, and Yanase that conservation laws limit the accuracy of "repeatable," or "nondisturbing," measurements to general measurements, and improves the one previously obtained by Yanase for spin measurements. The bound represents an obstacle to making a small quantum computer. PMID- 11863708 TI - Aharonov-Casher-effect suppression of macroscopic tunneling of magnetic flux. AB - We suggest a system in which the amplitude of macroscopic flux tunneling can be modulated via the Aharonov-Casher effect. The system is an rf SQUID with the Josephson junction replaced by a Bloch transistor--two junctions separated by a small superconducting island on which the charge can be induced by an external gate voltage. When the Josephson coupling energies of the junctions are equal and the induced charge is q = e, destructive interference between tunneling paths brings the flux tunneling rate to zero. The device may also be useful as a qubit for quantum computation. PMID- 11863709 TI - System size resonance in coupled noisy systems and in the Ising model. AB - We consider an ensemble of coupled nonlinear noisy oscillators demonstrating in the thermodynamic limit an Ising-type transition. In the ordered phase and for finite ensembles stochastic flips of the mean field are observed with the rate depending on the ensemble size. When a small periodic force acts on the ensemble, the linear response of the system has a maximum at a certain system size, similar to the stochastic resonance phenomenon. We demonstrate this effect of system size resonance for different types of noisy oscillators and for different ensembles-- lattices with nearest neighbors coupling and globally coupled populations. The Ising model is also shown to demonstrate the system size resonance. PMID- 11863710 TI - Quantum afterburner: improving the efficiency of an ideal heat engine. AB - By using a laser and maser in tandem, it is possible to obtain laser action in the hot exhaust gases of a heat engine. Such a "quantum afterburner" involves the internal quantum states of the working molecules as well as the techniques of cavity quantum electrodynamics and is therefore in the domain of quantum thermodynamics. It is shown that Otto cycle engine performance can be improved beyond that of the "ideal" Otto heat engine. Furthermore, the present work demonstrates a new kind of lasing without initial inversion. PMID- 11863711 TI - Observable dependence of fluctuation-dissipation relations and effective temperatures. AB - We study the nonequilibrium fluctuation-dissipation theorem (FDT) in the glass phase of Bouchaud's trap model. We incorporate an arbitrary observable m and obtain its correlation and response functions in closed form. A limiting nonequilibrium FDT plot is approached at long times for most choices of m. In contrast to standard mean field models, however, the shape of the plot depends nontrivially on the observable, and its slope varies continuously even though there is a single scaling of relaxation times with age. Nonequilibrium FDT plots can therefore not be used to define a meaningful effective temperature T(eff) in this model. Consequences for the wider applicability of an FDT-derived T(eff) are discussed. PMID- 11863713 TI - Determining the structure of Higgs couplings at the CERN LargeHadron Collider. AB - Higgs boson production via weak boson fusion at the CERN Large Hadron Collider has the capability to determine the dominant CP nature of a Higgs boson, via the tensor structure of its coupling to weak bosons. This information is contained in the azimuthal angle distribution of the two outgoing forward tagging jets. The technique is independent of both the Higgs boson mass and the observed decay channel. PMID- 11863712 TI - Measurements of cosmic-ray low-energy antiproton and proton spectra in a transient period of solar field reversal. AB - The energy spectra of cosmic-ray low-energy antiprotons ( *p's) and protons ( p's) have been measured by BESS in 1999 and 2000, during a period covering reversal at the solar magnetic field. Based on these measurements, a sudden increase of the *p/p flux ratio following the solar magnetic field reversal was observed, and it generally agrees with a drift model of the solar modulation. PMID- 11863714 TI - Measuring the photon polarization in B -->Kpipigamma. AB - We propose a way of measuring the photon polarization in radiative B decays into K resonance states decaying to Kpipi, which can test the standard model and probe new physics. The photon polarization is shown to be measured by the up-down asymmetry of the photon direction relative to the Kpipi decay plane in the K resonance rest frame. The integrated asymmetry in K1(1400)-->Kpipi, calculated to be 0.34 plus/minus 0.05 in the standard model, is measurable at currently operating B factories. PMID- 11863717 TI - Leading chiral contributions to the spin structure of the proton. AB - The leading chiral contributions to the quark and gluon components of the proton spin are calculated using heavy-baryon chiral perturbation theory. Similar calculations are done for the moments of the generalized parton distributions relevant to the quark and gluon angular momentum densities. These results provide useful insight into the role of pions in the spin structure of the nucleon and can serve as a guide for extrapolating lattice QCD calculations at large quark masses to the chiral limit. PMID- 11863718 TI - Intermediate mass fragment emission pattern in peripheral heavy-ion collisions at Fermi energies. AB - The emission pattern in the v(perp)-v(par) plane of intermediate mass fragments with Z = 3--7 (IMF) has been studied in the collision 116Sn+ 93Nb at 29.5A MeV as a function of the total kinetic energy loss of the reaction. This pattern shows that for peripheral reactions most IMF's are emitted at velocities intermediate between those of the projectile- and target-like products. Coulomb trajectory calculations show that these IMF's are produced in the interaction zone in a short time interval at the end of the target-projectile interaction. PMID- 11863719 TI - Local approximation of the correlation energy functional in the density matrix functional theory. AB - A local approximation formula of the correlation energy functional E(c) in terms of the first-order reduced density matrix (1-RDM) is presented. With the contracted Schrodinger equation the principal dependence of E(c) on the natural occupation numbers n(i) is identified. Using the effective mass theory, E(c) is expressed as a functional of the local density and the local variable, J = SUM (i)[square root of (n(i)(1-n(i))] /phi(i)/(2), where phi(i) are the natural spin orbitals. This local approximation satisfies the homogeneous coordinate scaling relation, gives the exact result for a one-electron system, and is almost free from the exchange energy error. It reproduced about 90% of the correlation energies of atoms and molecules. PMID- 11863720 TI - Magnetically quantized continuum distorted waves. AB - A new derivation of continuum distorted-wave theory is presented. It is generalized to magnetically quantized continuum distorted waves. The context is analytic continuation of hydrogenic-state wave functions from below to above threshold, using parabolic coordinates and quantum numbers including m the magnetic quantum number. This continuation applies to excitation, charge transfer, ionization, and double and hybrid events for both light- and heavy particle collisions. It is applied to the calculation of double-differential cross sections for the single ionization of the hydrogen atom and for a hydrogen molecule by a proton for electrons ejected in the forward direction at a collision energy of 50 keV and 100 keV respectively. PMID- 11863721 TI - Deceptive signals of phase transitions in small magnetic clusters. AB - We present an analysis of the thermodynamic properties of small transition-metal clusters and show how the commonly used indicators of phase transitions such as peaks in the specific heat or magnetic susceptibility can lead to deceptive interpretations of the underlying physics. The analysis of the distribution of zeros of the canonical partition function in the whole complex temperature plane reveals the nature of the transition. We show that signals in the magnetic susceptibility at positive temperatures have their origin at zeros lying at negative temperatures. PMID- 11863715 TI - Production of prompt charmonia in e(+)e(-) annihilation at square root of s approximately equals 0.6 GeV. AB - The production of prompt J/psi, psi(2S), chi(c1), and chi(c2) is studied using a 32.4 fb(-1) data sample collected with the Belle detector at Upsilon(4S) and at 60 MeV below the resonance. The yield of prompt J/psi mesons in the Upsilon(4S) sample is compatible with that of continuum production; we set an upper limit B(Upsilon(4S)-->J/psiX) < 1.9 x 10(-4) at the 95% confidence level, and find sigma(e(+)e(-)-->J/psiX) = 1.47 plus/minus 0.10 plus/minus 0.13 pb. The cross sections for prompt psi(2S) and direct J/psi are measured. The J/psi momentum spectrum, production angle distribution, and polarization are studied. PMID- 11863722 TI - Intrinsic and extrinsic nature of the orbital angular momentum of a light beam. AB - We explain that, unlike the spin angular momentum of a light beam which is always intrinsic, the orbital angular momentum may be either extrinsic or intrinsic. Numerical calculations of both spin and orbital angular momentum are confirmed by means of experiments with particles trapped off axis in optical tweezers, where the size of the particle means it interacts with only a fraction of the beam profile. Orbital angular momentum is intrinsic only when the interaction with matter is about an axis where there is no net transverse momentum. PMID- 11863716 TI - Observation of the color-suppressed decay B( 0)-->D(0)pi(0). AB - We report the first observation of color-suppressed B( 0)-->D(0)pi(0), D(*0)pi(0), D0eta, and D0omega decays, and evidence for B( 0)-->D(*0)eta and D(*0)omega. The branching fractions are B(B( 0)-->D0pi(0)) = (3.1 +/- 0.4 +/- 0.5)x10(-4), B(B( 0) -->D(*0)pi(0)) = (2.7(+0.8+0.5)(-0.7-0.6))x10(-4), B(B( 0) - > D0eta) = (1.4(+0.5)(-0.4) +/- 0.3)x10(-4), B(B( 0) --> D0omega) = (1.8 +/- 0.5(+0.4)(-0.3))x10(-4), and we set 90% confidence level upper limits of B(B( 0) -> D(*0)eta)<4.6 x 10(-4) and B(B( 0)-->D(*0)omega)<7.9 x 10(-4). The analysis is based on a data sample of 21.3 fb(-1) collected at the Upsilon(4S) resonance by the Belle detector at the KEKB e(+)e(-) collider. PMID- 11863723 TI - Coherent radiation from neutral molecules moving above a grating. AB - We predict and study the effect of parametric self-induced excitation of a molecule moving above the dielectric or conducting medium with periodic grating. In this case the radiation reaction force modulates the molecular transition frequency which results in a parametric instability of dipole oscillations even from the level of quantum or thermal fluctuations. The present mechanism of instability of electrically neutral molecules is different from that of the well known Smith-Purcell and transition radiation in which a moving charge and its oscillating image create an oscillating dipole. We show that parametrically excited molecular bunches can produce an easily detectable coherent radiation flux of up to a microwatt. PMID- 11863724 TI - Nonperturbative coherent population trapping: an analytic model. AB - Coherent population trapping is shown to occur in a driven symmetric double-well potential in the strong-field regime. The system parameters have been chosen to reproduce the 0(-) <--> 3(+) transition of the inversion mode of the ammonia molecule. For a molecule initially prepared in its lower doublet we find that, under certain circumstances, the 3(+) level remains unpopulated, and this occurs in spite of the fact that the laser field is resonant with the 0(-) <--> 3(+) transition and intense enough so as to strongly mix the 0(+) and 0(-) ground states. This counterintuitive result constitutes a coherent population trapping phenomenon of nonperturbative origin which cannot be accounted for with the usual models. PMID- 11863725 TI - Rotating optical soliton clusters. AB - We introduce the concept of soliton clusters--multisoliton bound states in a homogeneous bulk optical medium--and reveal a key physical mechanism for their stabilization associated with a staircaselike phase distribution that induces a net angular momentum and leads to cluster rotation. The ringlike soliton clusters provide a nontrivial generalization of the concepts of two-soliton spiraling, optical vortex solitons, and necklace-type optical beams. PMID- 11863726 TI - Semiquantal approach to finite systems of interacting particles. AB - A novel approach is suggested for the statistical description of quantum systems of interacting particles. We show that the occupation numbers for single-particle states can be represented as a convolution of a classical analog of the eigenstate, with the quantum occupation number for noninteracting particles. The latter takes into account the wave function symmetry and depends on the unperturbed energy spectrum only. As a result, the distribution of occupation numbers n(s) can be found even for a large number of interacting particles. Using the model of interacting spins, we demonstrate that this approach gives a correct description of n(s) even in deep quantum regions with few single-particle orbitals. PMID- 11863727 TI - Phase synchronization of chaotic rotators. AB - We demonstrate the existence of phase synchronization of two chaotic rotators. Contrary to phase synchronization of chaotic oscillators, here the Lyapunov exponents corresponding to both phases remain positive even in the synchronous regime. Such frequency locked dynamics with different ratios of frequencies are studied for driven continuous-time rotators and for discrete circle maps. We show that this transition to phase synchronization occurs via a crisis transition to a band-structured attractor. PMID- 11863728 TI - Sensitivity of wave field evolution and manifold stability in chaotic systems. AB - The sensitivity of a wave field's evolution to small perturbations is of fundamental interest. For chaotic systems, there are two distinct regimes of either exponential or Gaussian overlap decay in time. We develop a semiclassical approach for understanding both regimes and give a simple expression for the crossover time between the regimes. The wave field's evolution is considerably more stable than the exponential instability of chaotic trajectories seems to suggest. The resolution of this paradox lies in the collective behavior of the appropriate set of trajectories. Results are given for the standard map. PMID- 11863729 TI - Minimal model for sand dunes. AB - We propose a minimal model for aeolian sand dunes. It combines an analytical description of the turbulent wind velocity field above the dune with a continuum saltation model that allows for saturation transients in the sand flux. The model provides a qualitative understanding of important features of real dunes, such as their longitudinal shape and aspect ratio, the formation of a slip face, the breaking of scale invariance, and the existence of a minimum dune size. PMID- 11863730 TI - Large scale structures, symmetry, and universality in sandpiles. AB - We introduce a sandpile model where, at each unstable site, all grains are transferred randomly to downstream neighbors. The model is local and conservative, but not Abelian. This does not appear to change the universality class for the avalanches in the self-organized critical state. It does, however, introduce long-range spatial correlations within the metastable states. For the transverse direction d(perpendicular)>0, we find a fractal network of occupied sites, whose density vanishes as a power law with distance into the sandpile. PMID- 11863731 TI - Evolution of hexagonal patterns from controlled initial conditions in a Benard Marangoni convection experiment. AB - We report quantitative measurements of both wave number selection and defect motion in nonequilibrium hexagonal patterns. A novel optical technique ("thermal laser writing") is used to imprint initial patterns with selected characteristics in a Benard-Marangoni convection experiment. Initial patterns of ideal hexagons are imposed to determine the band of stable pattern wave numbers while initial patterns containing an isolated penta-hepta defect are imprinted to study defect propagation directions and velocities. The experimental results are compared to recent theoretical predictions. PMID- 11863732 TI - Controlling wake turbulence. AB - This Letter introduces a control strategy for taming the wake turbulence behind a cylinder. An angular momentum injection scheme is proposed to synchronize the vertical velocity field. We show that the base suction, wake formation length, absolute instability, and the Karman vortex street are effectively controlled by the angular momentum injection. A control equation is designed to implement the injection. The Navier-Stokes equations, along with the control equation, are solved. The occurrence of a new recirculation free zone is identified. PMID- 11863733 TI - Scaling and universality in turbulent convection. AB - Anomalous correlation functions of the temperature field in two-dimensional turbulent convection are shown to be universal with respect to the choice of external sources. Moreover, they are equal to the anomalous correlations of the concentration field of a passive tracer advected by the convective flow itself. The statistics of velocity differences is found to be universal, self-similar, and close to Gaussian. These results point to the conclusion that temperature intermittency in two-dimensional turbulent convection may be traced back to the existence of statistically preserved structures, as it is in passive scalar turbulence. PMID- 11863734 TI - Evolution of ultracold neutral plasmas. AB - We present the first large-scale simulations of an ultracold neutral plasma, produced by photoionization of laser-cooled xenon atoms, from creation to initial expansion, using classical molecular-dynamics methods with open boundary conditions. We reproduce many of the experimental findings such as the trapping efficiency of electrons with increased ion number, a minimum electron temperature achieved on approach to the photoionization threshold, and recombination into Rydberg states of an anomalously low principal quantum number. In addition, many of these effects establish themselves very early in the plasma evolution ( similar ns) before the present experimental observations begin. PMID- 11863735 TI - Simulation of the expansion of an ultracold neutral plasma. AB - We report the results of simulations that explain many properties of ultracold neutral plasmas. We find that three-body recombination is important at very low temperatures since it is a heating mechanism for the electron gas and it preferentially removes the slow ions from the plasma. We also find that collisions between cold electrons and Rydberg atoms are an important source of electron heating and deexcitation of atoms formed in the plasma. Simulations show that the Coulomb coupling constant does not become larger than similar1/5 for the reported experiments. PMID- 11863736 TI - Effect of hyperdiffusivity on turbulent dynamos with helicity. AB - In numerical studies of turbulence, hyperviscosity is often used as a tool to extend the inertial subrange and to reduce the dissipative subrange. By analogy, hyperdiffusivity (or hyperresistivity) is sometimes used in magnetohydrodynamics. The underlying assumption is that only the small scales are affected by this manipulation. In the present paper, possible side effects on the evolution of the large-scale magnetic field are investigated. It is found that for turbulent flows with helicity, hyperdiffusivity causes the dynamo-generated magnetic field to saturate at a higher level than normal diffusivity. This result is successfully interpreted in terms of magnetic helicity conservation, which also predicts that full saturation is reached only after a time comparable to the large-scale magnetic (hyper)diffusion time. PMID- 11863737 TI - Stochastic heating and acceleration of electrons in colliding laser fields in plasma. AB - We propose a mechanism that leads to efficient acceleration of electrons in plasma by two counterpropagating laser pulses. It is triggered by stochastic motion of electrons when the laser fields exceed some threshold amplitudes, as found in single-electron dynamics. It is further confirmed in particle-in-cell simulations. In vacuum or tenuous plasma, electron acceleration in the case with two colliding laser pulses can be much more efficient than with one laser pulse only. In plasma at moderate densities, such as a few percent of the critical density, the amplitude of the Raman-backscattered wave is high enough to serve as the second counterpropagating pulse to trigger the electron stochastic motion. As a result, even with one intense laser pulse only, electrons can be heated up to a temperature much higher than the corresponding laser ponderomotive potential. PMID- 11863739 TI - Observation of the fast potential change at L-H transition by a heavy-ion-beam probe on JFT-2M. AB - The fast potential change near the separatrix is measured directly at the L-H transition by a heavy-ion-beam probe. The potential changes with two different time scales at the L-H transition triggered by a sawtooth crash: it drops at first with the time scale of 10--100 mus just after the arrival of the heat pulse due to the sawtooth crash. Then, it decreases again at a few 100 mus after the first drop at a time scale of about 200 mus. PMID- 11863740 TI - Angular gaps in radial diffusion-limited aggregation: two fractal dimensions and nontransient deviations from linear self-similarity. AB - When suitably rescaled, the distribution of the angular gaps between branches of off-lattice radial diffusion-limited aggregation is shown to approach a size independent limit. The power-law expected from an asymptotic fractal dimension D = 1.71 arises only for very small angular gaps, which occur only for clusters significantly larger than M = 10(6) particles. Intermediate size gaps exhibit an effective dimension around 1.67, even for M--> infinity. They dominate the distribution for clusters with M<10(6). The largest gap approaches a finite limit extremely slowly, with a correction of order M(-0.17). PMID- 11863738 TI - Island dynamics in the large-helical-device plasmas. AB - In the Large Helical Device plasma discharges, the size of an externally imposed island with mode number ( n/m = 1/1) decreases substantially when the plasma is collisionless ( nu(*)< approximately 1) and the beta is finite ( > approximately 0.1%) at the island location. For the collisional plasmas with finite beta, on the other hand, the size of the island increases. However, there is a threshold in terms of the vacuum island size below which the island enlargement is not seen. PMID- 11863741 TI - Geometric approach to the dynamic glass transition. AB - We numerically study the potential energy landscape of a fragile glassy system and find that the dynamic crossover corresponding to the glass transition is actually the effect of an underlying geometric transition caused by the vanishing of the instability index of saddle points of the potential energy. Furthermore, we show that the potential energy barriers connecting local glassy minima increase with decreasing energy of the minima, and we relate this behavior to the fragility of the system. Finally, we analyze the real space structure of activated processes by studying the distribution of particle displacements for local minima connected by simple saddles. PMID- 11863742 TI - How to couple Landau theory to an equation of state. AB - We show how to construct a Landau-type free energy based on a primary order parameter coupled to finite strain with an elastic energy derived from an arbitrary equation of state V = V(P). The resulting class of models provides an excellent and efficient framework for the systematic study of phase transformations for a wide range of materials up to ultrahigh pressures. PMID- 11863744 TI - Evolution of gas-filled nanocracks in crystalline solids. AB - In this work, the evolution of gas-filled cracks under gas implantation and subsequent annealing is studied on the basis of an elastic continuum approach. The observed growth limitation of He-filled nanocracks in SiC is attributed to their stabilization by the formation of circular dislocation dipoles. The formation and Ostwald ripening of bubble-loop complexes at elevated temperatures is modeled in terms of gas atom exchange between such complexes coupled with local matrix atom exchange between bubbles and loops of the same complex. The scaling laws derived for the time dependence of bubble and loop sizes are found to be in good agreement with experimental data. PMID- 11863745 TI - Exact multifractal spectra for arbitrary laplacian random walks. AB - Iterated conformal mappings are used to obtain exact multifractal spectra of the harmonic measure for arbitrary Laplacian random walks in two dimensions. Separate spectra are found to describe scaling of the growth measure in time, of the measure near the growth tip, and of the measure away from the growth tip. The spectra away from the tip coincide with those of conformally invariant equilibrium systems with arbitrary central charge c < or = 1, with c related to the particular walk chosen, while the scaling in time and near the tip cannot be obtained from the equilibrium properties. PMID- 11863743 TI - Differential photoelectron holography: a new approach for three-dimensional atomic imaging. AB - We propose differential holography as a method to overcome the long-standing forward-scattering problem in photoelectron holography and related techniques for the three-dimensional imaging of atoms. Atomic images reconstructed from experimental and theoretical Cu 3p holograms from Cu(001) demonstrate that this method suppresses strong forward-scattering effects so as to yield more accurate three-dimensional images of side- and backscattering atoms. PMID- 11863746 TI - Submicrometer coherent neutron beam production using a thin-film waveguide. AB - We have experimentally demonstrated that planar neutron waveguides can be used as resonant beam couplers to efficiently produce a coherent neutron line source with cross sections in the submicrometer range. The Fraunhofer far-field diffraction pattern of the first three resonance modes was measured and found to be in excellent agreement with the theoretical model. Our measurements confirm that an excited exiting mode is fully coherent in the direction perpendicular to the surface of the thin-film coupler and may therefore be used for applications of interest to a broad user community in biochemical and semiconductor nanosciences such as static and time-resolved coherent speckle experiments or phase-contrast imaging. PMID- 11863747 TI - Reactions and diffusion of water and oxygen molecules in amorphous SiO2. AB - Water and oxygen molecules determine many of the properties of amorphous SiO2 used in several technologies, but the underlying atomic-scale processes remain unresolved. We report results of first-principles calculations showing that a wide range of behavior is possible in an amorphous environment, including diffusion of the molecule as a whole and various reactions with the network. Experimental data including oxygen exchange reaction and radiation sensitivity are accounted for. The possibility of H3O+ formation as a source of positive charge is discussed. PMID- 11863748 TI - Phase transitions in liquid crystals under negative pressures. AB - We report the first measurements of orientational order parameters and phase transition temperatures in nematic and smectic A liquid crystals under negative pressures generated by an isochoric cooling of small droplets embedded in a glass former. Comparison of isobaric and isochoric measurements allows us to estimate the coefficients coupling the order parameter and density of an extended Landau- de Gennes model of the nematic phase. PMID- 11863749 TI - Triple-point wetting on rough substrates. AB - The influence of substrate roughness on the wetting scenario of adsorbed van der Waals films is investigated by theory and experiment. Calculating the bending free energy penalty of a solid sheet picking up the substrate roughness, we show that a finite roughness always leads to triple-point wetting reducing the widths of the adsorbed solid films considerably as compared to that of smooth substrates. Testing the theory against our experimental data for molecular hydrogen adsorbed on gold, we find quantitative agreement. PMID- 11863750 TI - Anomaly of the nonergodicity parameter and crossover to white noise in the fast relaxation spectrum of a simple glass former. AB - We present quasielastic light scattering and dielectric spectra of the glass former alpha-picoline. At high temperatures the evolution of the susceptibility minimum is well described by the mode coupling theory (MCT). Below the critical temperature T(c) the simple scaling laws of MCT fail due to the appearance of the excess wing of the alpha process, which shows a universal evolution as a function of log(10)tau(alpha). Taking this into account, however, we observe the predicted cusplike anomaly of the nonergodicity parameter as well as a crossover to "white noise." PMID- 11863751 TI - Shear instability of gamma-Fe in bulk and in ultrathin films. AB - Using ab initio local-spin-density calculations we demonstrate that along the Bain path describing the transformation of face-centered-cubic (fcc) gamma-Fe into body-centered-cubic (bcc) alpha-Fe, tetragonal Fe is unstable against monoclinic shear deformations producing a nearly bcc structure. In the limit of a monolayer adsorbed on a fcc substrate, the epitaxial constraint suppresses the shear instability, but in ultrathin films with three to six monolayers a striped pattern of near-bcc domains develops, confirming recent observations by scanning tunneling microscopy. A strong correlation between the shear instability and the magnetic state is reported. PMID- 11863752 TI - Random walks on a ( 2+1)-dimensional deformable medium. AB - A model of random walks on a deformable medium is proposed in 2+1 dimensions. The behavior of the walk is characterized by the stability parameter beta and the stiffness exponent alpha. The average square end-to-end distance l approximately equals (2nu) and the average number of visited sites approximately equals (k) are calculated. As beta increases, for each alpha there exists a critical transition point beta(c) from purely random walks ( nu = 1/2 and k approximate to 1) to compact growth ( nu = 1/3 and k = 2/3). The relationship between beta(c) and alpha can be expressed as beta(c) = e(alpha). The landscape generated by a walk is also investigated by means of the visit number distribution N(n)(beta). There exists a scaling relationship of the form N(n)(beta)approximately n(-2)f(n/beta(z)). PMID- 11863753 TI - Measuring surface stress discontinuities in self-organized systems with X rays. AB - We have performed a grazing incidence x-ray diffraction study of the self organized N/Cu(001) system. Diffraction satellites associated with self organization are particularly intense around Bragg conditions of the bulk crystal. Bulk elastic relaxations due to surface stress discontinuities at domain boundaries are responsible for this feature. A quantitative analysis shows that these relaxations, computed by molecular dynamics or continuum elasticity, explain very well the whole diffraction study. A difference in surface stress of 7 N m(-1) between uncovered and N-covered regions of the Cu surface is shown to be the driving force for self-organization. PMID- 11863754 TI - Stability of metal vicinal surfaces revisited. AB - The stability of metal vicinal surfaces with respect to faceting is investigated using empirical potentials as well as electronic structure calculations. It is proven that for a wide class of empirical potentials all vicinal surfaces between (100) and (111) are unstable at 0 K when the role of third and farther nearest neighbors is negligible. However, electronic structure calculations reveal that the answer concerning the stability of vicinal surfaces is not so clear-cut. Finally, it is shown that surface vibrations at finite temperatures have little effect on the stability of vicinal surfaces. PMID- 11863755 TI - Stationary polaron motion in a polymer chain at high electric fields. AB - We study the stationary motion of a polaron in a conducting polymer in the presence of a high electric field. Using the Su-Schrieffer-Heeger model plus an electric field, we find that at polaron velocities not exceeding the sound velocity, the dissipation of the electronic energy into the lattice occurs via emission of phonons with single selected wave vector. For this case the corresponding contribution to the polaron mobility can be calculated analytically. We discuss the issue of the polaron stability with respect to dissociation in a very high field at supersonic velocities. PMID- 11863756 TI - Phase diagram of the one-dimensional extended Hubbard model at half filling. AB - We reexamine the ground-state phase diagram of the one-dimensional half-filled Hubbard model with on-site and nearest-neighbor repulsive interactions. We calculate second-order corrections to coupling constants in the weak-coupling renormalization-group approach ( g-ology) to show that the bond-charge-density wave (BCDW) phase exists for weak couplings in between the charge-density-wave (CDW) and spin-density-wave (SDW) phases. We find that the umklapp scattering of parallel-spin electrons destabilizes the BCDW state and gives rise to a bicritical point where the CDW-BCDW and SDW-BCDW continuous-transition lines merge into the CDW-SDW first-order transition line. PMID- 11863758 TI - Spin dynamics from time-dependent spin-density-functional theory. AB - We derive the spin-wave dynamics of a magnetic material from the time-dependent spin-density-functional theory in the linear response regime. The equation of motion for the magnetization includes, besides the static spin stiffness, a "Berry curvature" correction and a damping term. A gradient expansion scheme based on the homogeneous spin-polarized electron gas is proposed for the latter two quantities, and the first few coefficients of the expansion are calculated to second order in the Coulomb interaction. PMID- 11863757 TI - Effective mass anisotropy of gamma electrons in GaAs/AlGaAs quantum wells. AB - Resonant magnetotunneling in GaAs/Al(0.28)Ga0.72As double barrier structures is used to demonstrate that the effective mass of confined Gamma conduction electrons becomes anisotropic when an electric field is applied perpendicular to the interfaces. Although several authors have previously reported Gamma-related optical anisotropy, this is the first example of a corresponding electrical anisotropy. The results are explained using a quantum mechanical model involving interface band mixing that contains additional features not found in the optical case. PMID- 11863759 TI - Ab initio evaluation of the charge ordering in alpha'NaV2O5. AB - We report ab initio calculations of the charge ordering in alpha'NaV2O5 using large configurations interaction methods on embedded fragments. Our major result is that the 2p(y) electrons of the bridging oxygen of the rungs present a very strong magnetic character and should thus be explicitly considered in any relevant effective model. The most striking consequence of this result is that the spin and charge ordering differ substantially, as differ the experimental results depending on whether they are sensitive to the spin or charge density. PMID- 11863760 TI - Many-body GW calculations of ground-state properties: quasi-2D electron systems and van der Waals forces. AB - We present GW many-body results for ground-state properties of two simple but very distinct families of inhomogeneous systems in which traditional implementations of density-functional theory (DFT) fail drastically. The GW approach gives notably better results than the well-known random-phase approximation, at a similar computational cost. These results establish GW as a superior alternative to standard DFT schemes without the expensive numerical effort required by quantum Monte Carlo simulations. PMID- 11863761 TI - Entropy of vortex cores near the superconductor-insulator transition in an underdoped cuprate. AB - We present a study of Nernst effect in underdoped La(2-x)Sr(x)CuO4 in magnetic fields as high as 28 T. At high fields, a sizable Nernst signal was found to persist in the presence of a field-induced nonmetallic resistivity. By simultaneously measuring resistivity and the Nernst coefficient, we extract the entropy of vortex cores in the vicinity of this field-induced superconductor insulator transition. Moreover, the temperature dependence of the thermoelectric Hall angle provides strong constraints on the possible origins of the finite Nernst signal above T(c), as recently discovered by Xu et al. [Nature (London) 406, 486 (2000)]. PMID- 11863762 TI - Internal transitions of negatively charged magnetoexcitons and many body effects in a two-dimensional electron gas. AB - Internal transitions of quasi-two-dimensional, negatively charged magnetoexcitons ( X-) and their evolution with excess electron density have been studied in GaAs/AlGaAs quantum wells. In the dilute electron limit, due to magnetic translational invariance, the optically detected resonance spectra are dominated by bound-to-continuum bands in contrast to the negatively charged donor system D , which exhibits strictly bound-to-bound transitions. With increasing excess electron density Landau-level filling factors nu<2 the X--like transitions are blueshifted; they are absent for nu>2. The blueshifted transitions are explained in terms of a new type of collective excitation---magnetoplasmons bound to a mobile valence band hole. PMID- 11863763 TI - Reconstruction of fractional quantum Hall edges. AB - We study the interplay of electron-electron interaction, confining potential and effects of finite temperature at the edge of a quantum Hall liquid. Our exact diagonalization calculation indicates that edge reconstruction occurs in the fractional quantum Hall regime for a variety of confining potential, including ones that correspond to a "sharp" edge. Our finite temperature Hartree-Fock calculation for integer quantum Hall edges indicates that reconstruction is suppressed above a certain temperature. We discuss the implication of our results on recent edge tunneling and microwave absorption experiments. PMID- 11863764 TI - Electric polarization of heteropolar nanotubes as a geometric phase. AB - The threefold symmetry of planar boron nitride (BN), the III-V analog to graphene, prohibits an electric polarization in its ground state, but this symmetry is broken when the sheet is wrapped to form a BN nanotube. We show that this leads to an electric polarization along the nanotube axis which is controlled by the quantum mechanical boundary conditions on its electronic states around the tube circumference. Thus the macroscopic dipole moment has an intrinsically nonlocal quantum mechanical origin from the wrapped dimension. We formulate this novel phenomenon using the Berry's phase approach and discuss its experimental consequences. PMID- 11863765 TI - Electron holography of field-emitting carbon nanotubes. AB - Electron holography performed in situ inside a high resolution transmission electron microscope has been used to determine the magnitude and spatial distribution of the electric field surrounding individual field-emitting carbon nanotubes. The electric field (and hence the associated field emission current) is concentrated precisely at the tips of the nanotubes and not at other nanotube defects such as sidewall imperfections. The electric field magnitude and distribution are stable in time, even in cases where the nanotube field emission current exhibits extensive temporal fluctuations. PMID- 11863766 TI - Momentum-resolved lifetimes of image-potential States on cu(100). AB - The dependence of the inelastic lifetime of electrons in the image-potential states of Cu(100) on their momentum parallel to the surface has been studied experimentally by means of time- and angle-resolved two-photon photoemission and theoretically by evaluating the electron self-energy within the GW approximation. The pronounced decrease of the n = 1 lifetime from 40 fs at normal emission (k(parallel) = 0) to 20 fs for k(parallel) = 0.33 A(-1) cannot be accounted for by interband decay processes to bulk states. We show that intraband transitions within the image-state band give a contribution to this decrease comparable in magnitude with the interband channel. PMID- 11863767 TI - Ab initio investigation of collective charge excitations in MgB2. AB - A sharp collective charge excitation is predicted in MgB2 at approximately 2.5 eV for q perpendicular to the boron layers, based on an all-electron analysis of the dynamical density response within time-dependent density functional theory. This novel excitation, consisting of coherent charge fluctuation between Mg and B sheets, induces an abrupt plasma edge in the experimentally observable reflectivity. The existence of this mode reflects the unique electronic structure of MgB2 that is also responsible for strong electron-phonon coupling. By contrast, the acoustic plasmon, recently suggested to explain the high T(c), is not realized when realistic transition strengths are incorporated. PMID- 11863768 TI - Mechanism of pseudogap probed by a local impurity. AB - The response to a local strong nonmagnetic impurity in the pseudogap phase is examined in two distinctly different scenarios: phase fluctuation (PF) of pairing field and d-density-wave (DDW) order. In the PF scenario, the resonance state is generally double peaked near the Fermi level, and is abruptly broadened by vortex fluctuations slightly above the transition temperature. In the DDW scenario, the resonance is single peaked and remains sharp up to gradual intrinsic thermal broadening, and the resonance energy is analytically determined to be at minus of the chemical potential. PMID- 11863769 TI - Phase diagram and dynamics of the projected SO(5) symmetric model of high- T(c) superconductivity. AB - We present numerical studies of a quantum "projected" SO(5) model which aims at a unifying description of antiferromagnetism and superconductivity in the high- T(c) cuprates, while properly taking into account the Mott insulating gap. Our numerical results, obtained by the quantum Monte Carlo technique of stochastic series expansion, show that this model can give a realistic description of the global phase diagram of the high- T(c) superconductors and accounts for many of their physical properties. Moreover, we address the question of asymptotic restoring of the SO(5) symmetry at the critical point. PMID- 11863770 TI - d-Wave-induced Josephson current counterflow in YBa2Cu3O7/Nb zigzag junctions. AB - Well-defined zigzag-shaped ramp-type Josephson junctions between YBa2Cu3O7 and Nb have been studied. The magnetic field dependencies of the critical currents provide evidence for d-wave--induced alternations in the direction of the Josephson current between neighboring sides of the zigzag structure. The arrays present controllable model systems to study the influences of pi facets in high angle high- T(c) grain boundaries. From the characteristics, we estimate a possible imaginary s-wave admixture to the order parameter of the YBa2Cu3O7 to be below 1%. PMID- 11863771 TI - Spin-polarized scanning tunneling microscopy with antiferromagnetic probe tips. AB - We have performed low temperature spin-polarized scanning tunneling microscopy (SP-STM) of two monolayers Fe on W(110) using tungsten tips coated with different magnetic materials. We observe stripe domains with a magnetic period of 50 +/- 5 nm. Employing Cr as a coating material we recorded SP-STM images with an antiferromagnetic probe tip. The advantage of its vanishing dipole field is most apparent in external magnetic fields. This new approach resolves the problem of the disturbing influence of a ferromagnetic tip in the investigation of soft magnetic materials and superparamagnetic particles. PMID- 11863772 TI - Photoinduced magnetization in the organic-based magnet Mn(TCNE)(x)* y(CH2Cl2). AB - Photoinduced magnetization in a magnet based on organic species is reported for the first time. Upon optical excitation in the blue region of spectrum, Mn(tetracyanoethylene)(x)*y(CH2Cl2) (x approximately 2, y is approximately 0.8) exhibits increased magnetic susceptibility at temperatures as high as 75 K, accompanied with photoinduced absorption in the visible and infrared spectral regions. These effects are partially reversible by lower energy visible light and fully reversible by thermal treatment. The results suggest trapping of the photoexcited charge in a metastable state with enhanced exchange interaction. PMID- 11863773 TI - Analysis of a continuous field theory in two dimensions with use of the density matrix renormalization group. AB - A formulation of the Ginzburg-Landau-Wilson version of the partition function of a system with a continuously varying order parameter as a transfer matrix calculation allows for the application of methods based on the density matrix renormalization group (DMRG) to the calculation of the free energy of the O(1) model. The essence of both the mapping and the DMRG calculation is laid out, along with results that validate this strategy. PMID- 11863774 TI - Field-induced transitions in a kagome antiferromagnet. AB - The thermal order by disorder effect in magnetic field is studied for a classical Heisenberg antiferromagnet on the Kagome lattice. Using analytical arguments we predict a unique H- T phase diagram for this strongly frustrated magnet: states with a coplanar and a uniaxial triatic order parameter, respectively, at low and high magnetic fields and an incompressible collinear spin-liquid state at one third of the saturation field. We also present the Monte Carlo data which confirm the existence of these phases. PMID- 11863775 TI - Spin-sensitive bleaching and monopolar spin orientation in quantum wells. AB - Spin-sensitive bleaching of the absorption of far-infrared radiation has been observed in p-type GaAs/AlGaAs quantum well structures. The absorption of circularly polarized radiation saturates at lower intensities than that of linearly polarized light due to monopolar spin orientation in the first heavy hole subband. Spin relaxation times of holes in p-type material in the range of tens of ps were derived from the intensity dependence of the absorption. PMID- 11863776 TI - Ultrafast optical switching from an ionic to a neutral state in tetrathiafulvalene-p-chloranil (TTF-CA) observed in femtosecond reflection spectroscopy. AB - Ultrafast optical switching from an ionic ( I) to a neutral ( N) state in TTF-CA was observed in femtosecond reflection spectroscopy. Charge transfer excitation in the I phase produces N donor ( D0) acceptor ( A0) strings within 2 ps. These initial N states decay with a lifetime of 300 ps at 4 K, but rather multiply leading to macroscopic I-N conversion within 20 ps at 77 K just below the N-I transition temperature T(NI). Near T(NI), we also found the evidence for the coherent motion of the macroscopic N-I domain boundary with a period of 85 ps. PMID- 11863778 TI - Formation of a high T(c) electron-hole liquid in diamond. AB - We report on the observation of electron-hole ( e-h) liquid (EHL) in diamond by time-resolved luminescence measurements under an intense femtosecond photoexcitation above the band gap. The EHL luminescence band is observed below the e-h plasma band, showing a finite rise time of several tens of picoseconds. The rise time, which corresponds to the nucleation and the growth of the e-h droplets, plummets on approaching the EHL critical temperature. Time-resolved spectral shape analysis reveals a very high carrier density of 1x10(20) cm(-3) and very high critical temperature of T(c) = 165 K of EHL. PMID- 11863777 TI - Negative role of surface plasmons in the transmission of metallic gratings with very narrow slits. AB - It is generally admitted that the extraordinary transmission of metallic grating with very narrow slits is mainly due to the excitation of surface plasmons on the upper and lower interfaces of the grating. We show that the surface plasmon contribution is not the prime effect and that waveguide mode resonance and diffraction are responsible for the extraordinary transmission. Additionally and surprisingly, we reveal that the transmittance of subwavelength metallic gratings is always nearly zero for frequencies corresponding to surface plasmon excitation. This finding implies that surface plasmons play a negative role in the transmission. PMID- 11863779 TI - Electric quadrupolar contribution to the nuclear spin-lattice relaxation of Ir in Fe. AB - We report on the first quantitative determination of the electric quadrupolar contribution to the nuclear spin-lattice relaxation in a transition metal. For 186Ir and 189Ir in Fe we have determined the magnetic and the electric quadrupolar part of the relaxation for magnetic fields between 0.01 and 2 T. The quadrupolar part gives information on the role of the orbital motion of the electrons for the relaxation process. Our results prove that the unexpected high relaxation rates in Fe and their magnetic field dependence are due to a nonorbital relaxation mechanism. PMID- 11863780 TI - Jetlike component in sputtering of LiF induced by swift heavy ions. AB - Angular distributions of sputtered atoms from SiO2 and LiF single crystals were measured under the irradiation of 1 MeV/u swift heavy ions. In contrast to the almost isotropic distribution of SiO2, an additional jetlike component was observed for LiF. The total sputtering yield of SiO2 ( approximately 10(2) atoms/ion) can be reproduced by an extended inelastic thermal spike model, whereas the huge yield of LiF ( approximately 10(4) atoms/ion) needs a substantial decrease of the sublimation energy to be described by the model. PMID- 11863781 TI - Qubit channels can require more than two inputs to achieve capacity. AB - We give examples of qubit channels that require three input states in order to achieve the Holevo capacity. PMID- 11863782 TI - Continuous variable quantum cryptography using coherent states. AB - We propose several methods for quantum key distribution (QKD) based on the generation and transmission of random distributions of coherent or squeezed states, and we show that they are secure against individual eavesdropping attacks. These protocols require that the transmission of the optical line between Alice and Bob is larger than 50%, but they do not rely on "sub-shot noise" features such as squeezing. Their security is a direct consequence of the no-cloning theorem, which limits the signal-to-noise ratio of possible quantum measurements on the transmission line. Our approach can also be used for evaluating various QKD protocols using light with Gaussian statistics. PMID- 11863783 TI - Spiral wave generation in heterogeneous excitable media. AB - As the coupling in a heterogeneous excitable medium is reduced, three different types of behavior are encountered: plane waves propagate without breaking up, plane waves break up into spiral waves, and plane waves block. We illustrate these phenomena in monolayers of chick embryonic heart cells using calcium sensitive fluorescent dyes. Following the addition of heptanol, an agent that reduces the electrical coupling between cells, we observe breakup of spiral waves. These results are modeled in a heterogeneous cellular automaton model in which the neighborhood of interaction is modified. PMID- 11863784 TI - Molecular observation of contour-length fluctuations limiting topological confinement in polymer melts. AB - In order to study the mechanisms limiting the topological chain confinement in polymer melts, we have performed neutron-spin-echo investigations of the single chain dynamic-structure factor from polyethylene melts over a large range of chain lengths. While at high molecular weight the reptation model is corroborated, a systematic loosening of the confinement with decreasing chain length is found. The dynamic-structure factors are quantitatively described by the effect of contour-length fluctuations on the confining tube, establishing this mechanism on a molecular level in space and time. PMID- 11863785 TI - Comparison of nonequilibrium molecular dynamics with experimental measurements in the nonlinear shear-thinning regime. AB - Nonequilibrium molecular dynamics of a low-molecular-weight fluid (squalane) are compared with experimental measurements in both the linear (Newtonian) and nonlinear (non-Newtonian) regimes. The experimental and simulation data are shown to follow the same time-temperature superposition master curve. This represents the first comparison of the nonlinear rheology predicted by nonequilibrium molecular dynamics with experiment, and is thus the first experimental test of nonequilibrium molecular dynamics simulations in the nonlinear regime. PMID- 11863786 TI - Comment on "Evidence of a sponge-to-lamellar phase transition under shear by X ray scattering experiments in a couette cell". PMID- 11863790 TI - Comment on "Bicritical and tetracritical phenomena and scaling properties of the SO(5) theory". PMID- 11863788 TI - Comment on "Spinless impurities in high- T(c) cuprates: Kondo-like behavior". PMID- 11863792 TI - Spin waves in a Bose-Einstein--condensed atomic spin chain. AB - The spin dynamics of atomic Bose-Einstein condensates confined in a one dimensional optical lattice is studied. The condensates at each lattice site behave like spin magnets that can interact with each other through both the light induced dipole-dipole interaction and the static magnetic dipole-dipole interaction. We show how these site-to-site dipolar interactions can distort the ground-state spin orientations and lead to the excitation of spin waves. The dispersion relation of the spin waves is studied and possible detection schemes are proposed. PMID- 11863793 TI - Experimental observation of the Bogoliubov transformation for a Bose-Eeinstein condensed gas. AB - Phonons with wave vector q/(planck constant) were optically imprinted into a Bose Einstein condensate. Their momentum distribution was analyzed using Bragg spectroscopy with a high momentum transfer. The wave function of the phonons was shown to be a superposition of +q and -q free particle momentum states, in agreement with the Bogoliubov quasiparticle picture. PMID- 11863794 TI - Violating Bell's inequality beyond Cirel'son's bound. AB - Cirel'son inequality states that the absolute value of the combination of quantum correlations appearing in the Clauser-Horne-Shimony-Holt (CHSH) inequality is bound by 2 square root of (2). It is shown that the correlations of two qubits belonging to a three-qubit system can violate the CHSH inequality beyond 2 square root of (2). Such a violation is not in conflict with Cirel'son's inequality because it is based on postselected systems. The maximum allowed violation of the CHSH inequality, 4, can be achieved using a Greenberger-Horne-Zeilinger state. PMID- 11863795 TI - Dielectric black hole analogs. AB - As an alternative to the sonic black hole analogs we discuss a different scenario for modeling the Schwarzschild geometry in a laboratory--the dielectric black hole. The dielectric analog of the horizon occurs if the velocity of a medium with a finite permittivity exceeds the speed of light in that medium. The relevance for experimental tests of the Hawking effect and possible implications are addressed. PMID- 11863796 TI - Lambda(0) polarization in 800-GeV/cpp --> p(f)(Lambda0K+). AB - We report results from a study of Lambda(0) polarization in the exclusive reaction pp --> p(f)(Lambda0K+) at 800-GeV/c. We observe a dependence of the polarization on the Lambda0K+ invariant mass with large (+71%) positive polarization at small mass (1.63-GeV/c(2)) and large (-43%) negative polarization at large mass (2.75-GeV/c(2)). This observation confirms the result of the CERN ISR R608 experiment and extends the range over which the effect is observed. The strong dependence of the polarization on the Lambda0K+ invariant mass suggests that the origin of the polarization is closely related to the production dynamics of the diffractively produced Lambda0K+ system. PMID- 11863799 TI - Second-order dissipative fluid dynamics for ultrarelativistic nuclear collisions. AB - The Muller-Israel-Stewart second-order theory of relativistic imperfect fluids based on Grad's moment method is used to study the expansion of hot matter produced in ultrarelativistic heavy-ion collisions. The temperature evolution is investigated in the framework of the Bjorken boost-invariant scaling limit. The results of these second-order theories are compared to those of first-order theories due to Eckart and to Landau and Lifshitz and those of zeroth order (perfect fluid) due to Euler. PMID- 11863798 TI - Lambda spectra in 11.6A GeV/c Au-Au collisions. AB - E896 has measured Lambda production in 11.6A GeV/c Au-Au collisions over virtually the whole rapidity phase space. The midrapidity p(t) distributions have been measured for the first time at this energy and appear to indicate that the Lambda hyperons have different freeze-out conditions than protons. A comparison with the relativistic quantum molecular dynamics model shows that while there is good shape agreement at high rapidity the model predicts significantly different slopes of the m(t) spectra at midrapidity. The data, where overlap occurs, are consistent with previously reported measurements. PMID- 11863801 TI - Neutron-induced fission cross section of 233Pa between 1.0 and 3.0 MeV. AB - The energy dependent neutron-induced fission cross section of 233Pa has for the first time been measured directly with monoenergetic neutrons. This nuclide is an important intermediary in a thorium based fuel cycle, and its fission cross section is a key parameter in the modeling of future advanced fuel and reactor concepts. A first experiment resulted in four cross section values between 1.0 and 3.0 MeV, establishing a fission threshold in excess of 1 MeV. Significant discrepancies were found with a previous indirect experimental determination and with model estimates. PMID- 11863800 TI - Electrostatic mapping of nuclear pairing. AB - The traditional nuclear pairing problem is shown to be in one-to-one correspondence with a classical electrostatic problem in two dimensions. We make use of this analogy in a series of calculations in the tin region, showing that the extremely rich phenomenology that appears in this classical problem can provide interesting new insights into nuclear superconductivity. PMID- 11863802 TI - Observation of magnetoelectric linear birefringence. AB - We report the first experimental observation of optical linear birefringence induced in molecular liquids by crossed electric and magnetic fields perpendicular to the direction of light propagation. The optical axes coincide with the external fields, and the strength is bilinear in the electric and magnetic fields. PMID- 11863797 TI - Observation of B(0) --> D0pi(0) and B(0) -->D(*0)pi(0). AB - We have studied the color-suppressed hadronic decays of neutral B mesons into the final states D*0pi(0). Using 9.67 x 10(6) BB pairs collected with the CLEO detector, we observe the decays B( 0) --> D0pi(0) and B( 0) -->D(*0)pi(0) with the branching fractions BB( 0) -->D0pi(0)) = (2.74(+0.36)(-0.32) +/- 0.55)x10(-4) and BB( 0) -->D(*0)pi(0)) = (2.20(+0.59)(-0.52) +/- 0.79)x10(-4). The first error is statistical and the second systematic. The statistical significance of the D0pi(0) signal is 12.1sigma ( 5.9sigma for D(*0)pi(0)). Utilizing the B( 0) - >D*0)pi(0) branching fractions we determine the strong phases delta(I,D(*)) between isospin 1/2 and 3/2 amplitudes in the Dpi and D*pi final states to be cosdelta(I,D) = 0.89 +/-0.08 and cosdelta(I,D*) = 0.89 +/- 0.08, respectively. PMID- 11863803 TI - Resonant coupling of free quantum rotors in inclusion compounds. AB - NH3 groups in certain Hofmann clathrates form almost free one-dimensional quantum rotors with energy levels E(n) = n(2)B and angular momentum n (planck constant), where n = 0, +/-1, +/-2,.... Recent neutron scattering experiments revealed a surprising temperature dependence for the linewidths of the n = 0 <--> 1, 0 <--> 2, and 1 <--> 2 transitions. We propose a novel line broadening mechanism based on rotor-rotor coupling and obtain a simple analytic expression for the widths that depends on the rotor level occupation and on the 3-proton spin degeneracies of initial and final states. Our model provides, without adjustable parameters, a good fit both to the temperature dependence of the observed widths and to their relative magnitude. PMID- 11863805 TI - Narrow-band coherent anti-stokes Raman signals from broad-band pulses. AB - By tailoring the phase of a 100 femtosecond probe pulse we are able to obtain a narrow-band coherent anti-Stokes Raman spectroscopy (CARS) resonant signal with a width of less than 15 cm(-1), which is an order of magnitude narrower than the CARS signal from a transform limited pulse. Thus, by measuring the spectrum of the CARS signal we are able to obtain a high-resolution energy level diagram of the probed sample in spite of the broad femtosecond pulse spectrum. PMID- 11863804 TI - Direct probe of the shape resonance mechanism in 2sigma(g)-shell photoionization of the N2 molecule. AB - Angular distributions of photoelectrons from a 2sigma(g) shell of fixed-in-space N2 molecules have been measured for left- and right-elliptically polarized and for linearly polarized light at several photon energies in the region of sigma(*) shape resonance. That allowed the determination of a set of dipole matrix elements and phase shift differences characterizing the process. These data clearly show the enhancement of the fsigma(u) partial cross section in the resonance simultaneously with an abrupt increase of the corresponding phase shift by pi, which is the first experimental demonstration of the nature of the sigma(*) shape resonance in homonuclear diatomic molecules. PMID- 11863806 TI - Induced long-range dipole-field-enhanced antihydrogen formation in the p + Ps(n = 2) --> e(-) + H(n < or = 2) reaction. AB - We report high-precision calculations that correctly include the rearrangement channels by solving the modified Faddeev equations for energies between the Ps(n = 2) and H(n = 3) thresholds, which involve six and eight open channels. We find that 99% of the antihydrogen is formed in H(n = 2). Just above the Ps(n = 2) threshold the S, P, and D partial waves contribute nearly 1400pi(a(2)0) near the maximum. We find evidence that the induced long-range dipole potential is responsible for such a large H formation cross section. The possibility of utilizing this resonance to synthesize low-energy H is discussed. PMID- 11863807 TI - Manipulation of photon statistics of highly degenerate incoherent radiation. AB - Highly degenerate incoherent radiation has a Gaussian density matrix and a large occupation number of modes f. If it is passed through a weakly transmitting barrier, its counting statistics is close to Poissonian. We show that a second identical barrier, in series with the first, drastically modifies the statistics. The variance of the photocount is increased above the mean by a factor f times a numerical coefficient. The photocount distribution reaches a limiting form with a Gaussian body and highly asymmetric tails. These are general consequences of the combination of weak transmission and multiple scattering. PMID- 11863808 TI - Multipulse excitability in a semiconductor laser with optical injection. AB - An optically injected semiconductor laser can produce excitable multipulses. Homoclinic bifurcation curves confine experimentally accessible regions in parameter space where the laser emits a certain number of pulses after being triggered from its steady state by a single perturbation. This phenomenon is organized by a generic codimension-two homoclinic bifurcation and should also be observable in other systems. PMID- 11863810 TI - Coexistence of two granular temperatures in binary vibrofluidized beds. AB - An investigation into the granular temperature distributions of a binary vibrofluidized granular bed has been conducted using positron emission particle tracking. By repeating each experiment with the tracer selected in turn from the two size components, the granular temperature and packing fraction distributions for each phase were determined. It was found that, for a range of size fractions, the granular temperature of the larger particles was higher than that of the smaller diameter grains, a result which was supported by a simple theoretical analysis based on the steady state energy equation. PMID- 11863809 TI - Experimental observation of wave chaos in a conventional optical resonator. AB - We report on an experimental observation of optical wave chaos in a resonator consisting of three standard, high-reflectivity mirrors. The nonseparability of the wave equation necessary for chaos is introduced by violating the paraxial approximation. Until recently progress in optical wave chaos was hampered by the inherent difficulty in realizing suitable microscopic systems; now this novel, macroscopic approach offers complete and easy control and allows unprecedented study of optical wave chaos. PMID- 11863811 TI - Heat-flux measurement in high-Prandtl-number turbulent Rayleigh-Benard convection. AB - We report Nusselt number measurements from high Prandtl number turbulent thermal convection experiments. The experiments are conducted in four fluids with the Prandtl number Pr varying from 4 to 1350 and the Rayleigh number Ra from 2x10(7) to 3x10(10), all in a single convection cell of unity aspect ratio. We find that the measured Nusselt number decreased about 20% over the range of Pr spanned in the experiment. The measure data are also found in good agreement with the prediction of a recent theory over the extended range of Pr covered in the experiment. PMID- 11863812 TI - Effect of vorticity production on the structure and velocity of curved flames. AB - A nonlinear equation describing curved stationary flames with arbitrary gas expansion theta = rho(fuel)/rho(burnt) is obtained in a closed form without an assumption of weak nonlinearity. The equation respects all conservation laws and takes into account vorticity production in the flame. In the scope of the asymptotic expansion for theta -->1, the new equation solves the problem of stationary flame propagation with accuracy of the sixth order in theta - 1. Its analytical solutions give the flame velocity in tubes of arbitrary width, which agrees with available results of direct numerical modeling. PMID- 11863813 TI - Free-electron RF-pulse compressor. AB - A spatially modulated (corrugated) electron beam provides Bragg-type scattering of two waveguide modes. This allows the use of corrugated beams as quick active elements in rf-pulse compressors. In an illustrative example shown with a peak power compression ratio of 20-40, a 10-20 ns output pulse with a peak power of tens of MW is predicted for a millimeter-wavelength compressor. PMID- 11863814 TI - Optimal wire-number range for high x-ray power in long-implosion-time aluminum Z pinches. AB - Experiments performed on the 8-MA Saturn accelerator to investigate the effects of interwire gap spacing on long-implosion-time Z pinches have resulted in the observation of a regime of optimal wire number. The experiments varied the wire number of 40 and 32 mm diam arrays, resulting in interwire gaps from 3.9 to 0.36 mm, with fixed mass and length. aluminum K-shell powers up to 3.4 TW were measured, with long, slow rising, lower power x-ray pulses for interwire gaps greater than 2.2 mm and less than 0.7 mm, and short, fast rising, higher power pulses for interwire gaps in the range 0.7-2.2 mm. PMID- 11863815 TI - Transport coefficients of the Yukawa one-component plasma. AB - We present equilibrium molecular-dynamics computations of the thermal conductivity and the two viscosities of the Yukawa one-component plasma. The simulations were performed within periodic boundary conditions, and Ewald sums were implemented for the potentials, the forces, and for all the currents which enter the Kubo formulas. For large values of the screening parameter, our estimates of the shear viscosity and the thermal conductivity are in good agreement with the predictions of the Chapman-Enskog theory. PMID- 11863816 TI - Numerical simulation of ultracold plasmas: how rapid intrinsic heating limits the development of correlation. AB - In recent experiments, ultracold plasmas were produced by photoionizing small clouds of laser-cooled atoms. It has been suggested that the low initial temperature of these novel plasmas leads directly to strong correlation and order. In contrast, we argue that rapid intrinsic heating raises the electron temperature to the point where strong correlation cannot develop. The argument is corroborated by a molecular-dynamics simulation of the early-time plasma evolution. PMID- 11863817 TI - Transitions from vortex lines to sheets: interplay of topology and dynamics in an anisotropic superfluid. AB - In isotropic macroscopic quantum systems vortex lines can be formed while in anisotropic systems also vortex sheets are possible. Based on measurements of superfluid 3He-A, we present the principles which select between these two competing forms of quantized vorticity: sheets displace lines if the frequency of the external drive exceeds a critical limit. The resulting topologically stable state consists of multiple vortex sheets and has much faster dynamics than the state with vortex lines. PMID- 11863818 TI - Ferromagnetism of 3He films in the low-field limit. AB - We provide evidence for a finite-temperature ferromagnetic transition in two dimensions as H -->0 in thin films of 3He on graphite, a model system for the study of two-dimensional magnetism. We perform pulsed and cw NMR experiments at fields of 0.03-0.48 mT on 3He at areal densities of 20.5-24.2 atoms/nm(2). At these densities, the second layer of 3He has a strongly ferromagnetic tendency. With decreasing temperature, we find a rapid onset of magnetization that becomes independent of the applied field at temperatures in the vicinity of 1 mK. Both the dipolar field and the NMR linewidth grow rapidly as well, which is consistent with a large (order unity) polarization of the 3He spins. PMID- 11863819 TI - Kinetic theory of symmetry-dependent strength in carbon nanotubes. AB - Carbon nanotubes yield to mechanical force by a primary dislocation dipole whose formation energy describes the thermodynamic stability of the tubule. However, the real-time strength is determined by the rate of defect formation, defined in turn by the activation barrier for the bond flip. First extensive computations of the kinetic barriers for a variety of strain-lattice orientations lead to predictions of the yield strength. Its value depends on nanotube chiral symmetry, in a way very different from the thermodynamic assessment. PMID- 11863820 TI - Chiral polymer hexatics: a new twist on DNA. AB - Theories of the " N+6" hexagonally ordered phase of DNA are generally based on treatments of hexatic order in smectic systems. Thus N+6 phases should be analogous to superconductors and should expel twist of the nematic director, in an analogy of the Meissner effect. However, in N+6 systems there is no smectic order, hence a radically different mathematical description is required. Here I present the appropriate theory and show that, as N+6 phases are not analogous to superconductors, they can exhibit twist in any direction, without forming any topological defects. PMID- 11863821 TI - Cooperative nucleation of shear dislocation loops. AB - The mean elastic interaction between randomly distributed transient subcritical shear loops of the same sign, formed in the presence of an applied shear stress, is the "image stress." This stress is proportional to the volume density of loops and has the same sign as the applied stress. The image stress promotes the cooperative nucleation of shear loops, and leads to an instability in which the number of loops and the image stress increase rapidly, leading to the generation of stable expanding loops. The critical stress at which the instability is predicted is relatively high. PMID- 11863822 TI - Induced anticlinic ordering and nanophase segregation of bow-shaped molecules in a smectic solvent. AB - Recent experiments indicate that doping low concentrations of bent-core molecules into calamitic smectic solvents can induce anticlinic and biaxial smectic phases. We have carried out Monte Carlo simulations of mixtures of rodlike molecules (hard spherocylinders with length/breadth ratio L(rod)/D = 5) and bow-shaped molecules (hard spherocylinder dimers with length/breadth ratio L(ban)/D = 5 or 2.5 and opening angle psi). We find that a low concentration ( 3%) of L(ban)/D = 5 dimers induces anticlinic ( SmC(A)) ordering in an untilted smectic ( SmA) phase for 100 < or = psi < 150. For L(ban)/D = 2.5, no tilted phases are induced. However, with decreasing psi we observe a sharp transition from intralamellar nanophase segregation (bow-shaped molecules segregated within smectic layers) to interlamellar nanophase segregation (bow-shaped molecules concentrated between smectic layers) near psi = 130. PMID- 11863823 TI - Observation of Kossel and Kikuchi lines in thermal neutron incoherent scattering. AB - In this Letter we report the observation of K lines (representing collectively, Kossel and Kikuchi lines) produced by monochromatic thermal neutrons interacting with a KDP (potassium dihydrogen phosphate) single crystal. Since K lines contain phase information, these observations establish the experimental basis for direct crystallographic phasing of atomic structures containing incoherent scatterers, such as hydrogen, via thermal neutron "inside source" holography. PMID- 11863824 TI - Free volume and density and temperature dependence of diffusion coefficients of liquid mixtures. AB - A simple formula for the diffusion coefficient of liquid mixtures, expressed in terms of the work necessary to create a characteristic free volume in the liquid, is presented in the spirit of the Arrhenius activation theory and tested in comparison with available experimental data. If use is made of the generic van der Waals equation of state, the free volume appearing in the formula for the diffusion coefficient can be expressed in terms of the equilibrium pair correlation functions. The theoretical values for diffusion coefficients agree excellently with experimental values with regard to the density and temperature dependence of the diffusion coefficients of argon and krypton. PMID- 11863825 TI - Spontaneous assembly of perfectly ordered identical-size nanocluster arrays. AB - A method, by which periodic two-dimensional arrays of identical metal clusters of nanometer size and spacing could be spontaneously obtained by taking advantage of surface mediated clustering, is reported. The versatility of the method is demonstrated for a broad range of metals on Si(111)-(7 x 7) substrates. In situ scanning tunneling microscopy analysis of In clusters, combined with first principles total energy calculations, unveils unique initial-stage atomic structures of the surface-supported clusters and the vital steps that lead to the success of this method. A strong interaction between the clusters and the surface holds the key to the observed cluster sizes. PMID- 11863826 TI - Intermolecular bond length of ice on Ag(111). AB - Water adsorbed in submonolayer coverage on Ag(111) at 70 K forms hydrogen-bonded networks. High resolution images in combination with calculation reveal that single protrusions represent a cyclic water hexamer with the intermolecular bond stretched to the silver lattice constant of 0.29 nm. Scanning tunneling spectroscopy indicates that the bond length within the two-dimensional hydrogen bonded water layer is shortened. The spectra contain further information about the vibrational modes of water molecules. PMID- 11863827 TI - First-principles surface phase diagram for hydrogen on GaN surfaces. AB - We discuss the derivation and interpretation of a generalized surface phase diagram, based on first-principles density-functional calculations. Applying the approach to hydrogenated GaN surfaces, we find that the Gibbs free energies of relevant reconstructions strongly depend on temperature and pressure. Choosing chemical potentials as variables results in a phase diagram that provides immediate insight into the relative stability of different structures. A comparison with recent experiments illustrates the power of the approach for interpreting and predicting energetic and structural properties of surfaces under realistic growth conditions. PMID- 11863828 TI - Field-dependent anisotropy change in a supramolecular Mn(II)- [3 x 3] grid. AB - The magnetic anisotropy of a novel Mn(II)- [3x3] grid complex was investigated by means of high-field torque magnetometry. Torque vs field curves at low temperatures demonstrates a ground state with S>0 and exhibits a torque step due to a field-induced level crossing at B(*) approximately 7.5 T, accompanied by an abrupt change of magnetic anisotropy from easy-axis to hard-axis types. These observations are discussed in terms of a spin Hamiltonian formalism. PMID- 11863829 TI - Shallow donor state due to nitrogen-hydrogen complex in diamond. AB - Based on an ab initio calculation, we propose a possible shallowing of a nitrogen (N) donor in diamond, in contrast to the traditional thinking that it is very deep. A complex defect of N and hydrogen (H), N-H-N, should be much shallower than an isolated N donor. A qualitative scenario for formation of the N-H-N defects is presented. The existence of this complex is strongly suggested by a recent discovery of a new muonium center in N-rich diamond. PMID- 11863830 TI - Vertex corrections near the stripe phase. AB - We calculate the vertex corrections within a model for fermion quasiparticles coupled with charge and spin fluctuations, which provide the relevant scattering mechanism near the stripe instability in high- T(c) cuprates. The logarithmic divergence of the vertex, which characterizes the spin-fermion model near the antiferromagnetic instability, is ruled out, due to the incommensuration of the charge and spin modulation within the stripe phase, as revealed by neutron scattering. This simplifies the skeleton structure of the problem. The vertex is negative in the relevant kinematical regime, effectively reducing the interaction strength. Our results apply to generic incommensurate instabilities of electronic origin. PMID- 11863831 TI - Excitonic effects in solids described by time-dependent density-functional theory. AB - Starting from the many-body Bethe-Salpeter equation we derive an exchange correlation kernel f(xc) that reproduces excitonic effects in bulk materials within time-dependent density functional theory. The resulting f(xc) accounts for both self-energy corrections and the electron-hole interaction. It is static, nonlocal, and has a long-range Coulomb tail. Taking the example of bulk silicon, we show that the -alpha/q(2) divergency is crucial and can, in the case of continuum excitons, even be sufficient for reproducing the excitonic effects and yielding excellent agreement between the calculated and the experimental absorption spectrum. PMID- 11863832 TI - Bipolar doping and band-gap anomalies in delafossite transparent conductive oxides. AB - Doping wide-gap materials p type is highly desirable but often difficult. This makes the recent discovery of p-type delafossite oxides, CuM(III)O2, very attractive. The CuM(III)O2 also show unique and unexplained physical properties: Increasing band gap from M(III) = Al,Ga, to In, not seen in conventional semiconductors. The largest gap CuInO2 can be mysteriously doped both n and p type but not the smaller gaps CuAlO2 and CuGaO2. Here, we show that both properties are results of a large disparity between the fundamental gap and the apparent optical gap, a finding that could lead to a breakthrough in the study of bipolarly dopable wide-gap semiconductor oxides. PMID- 11863833 TI - Spin filtering of free electrons by magnetic multilayers: towards an efficient self-calibrated spin polarimeter. AB - An asymmetrical ferromagnetic cobalt bilayer (18 nm Au/0.8 nm Co/2.2 nm Au/1.3 nm Co/1.5 nm Au) operates as a self-calibrated spin polarimeter with a high spin selectivity for free electrons injected at a few eV above the Fermi level. We present the analysis of transmitted currents as a function of the incident energy, based on a model of spin polarization dilution into the first gold layer and ballistic transport close to the vacuum level throughout the sample. PMID- 11863834 TI - Band-gap tuning and linear magnetoresistance in the silver chalcogenides. AB - Optimally doped silver selenide and silver telluride exhibit linear positive magnetoresistance over decades in magnetic field and on a scale comparable to the colossal magnetoresistance compounds. We use hydrostatic pressure to smoothly alter the band structure of Ag-rich and Ag-deficient samples of semiconducting Ag(2 +/-delta)Te of fixed stoichiometry and disorder. We find that the magnetoresistance spikes and the linear field dependence emerges when the bands cross and the Hall coefficient changes sign. PMID- 11863835 TI - Spin-polarized transport in inhomogeneous magnetic semiconductors: theory of magnetic/nonmagnetic p-n junctions. AB - A theory of spin-polarized transport in inhomogeneous magnetic semiconductors is developed and applied to magnetic/nonmagnetic p-n junctions. Several phenomena with possible spintronic applications are predicted, including spin-voltaic effect, spin valve effect, exponential and giant magnetoresistance. It is demonstrated that only nonequilibrium spin can be injected across the space charge region of a p-n junction, so that there is no spin injection (or extraction) at low bias. PMID- 11863836 TI - Experimental demonstration of Fermi surface effects at filling factor 5/2. AB - Using small wavelength surface acoustic waves (SAW) on ultrahigh mobility heterostructures, Fermi surface properties are detected at 5/2 filling factor at temperatures higher than those at which the quantum Hall state forms. An enhanced conductivity is observed at 5/2 by employing sub-0.5-microm SAW, indicating a quasiparticle mean-free path substantially smaller than that in the lowest Landau level. These findings are consistent with the presence of a filled Fermi sea of composite fermions, which may pair at lower temperatures to form the 5/2 ground state. PMID- 11863837 TI - Spectroscopic and magnetic mirages of impurities in nanoscopic systems. AB - We present exact results for magnetic impurities in nanoscopic systems with focusing properties. We analyze the spectroscopic and magnetic properties of Kondo, intermediate valence, and magnetic impurities on a sphere with a metallic surface. Exact calculations show the occurrence of spectroscopic and magnetic mirages at the antipodes of the impurity location. Comparison with calculations performed using effective models validates previous calculations of spectroscopic mirages. Our results predict the existence of a strong magnetic mirage in the experimentally realizable elliptic corral. PMID- 11863838 TI - Spin-polarized tunneling spectroscopy of co(0001) surface States. AB - Spin-polarized tunneling was studied on Co surfaces of exchange coupled Co/Cu/Co samples, using an Fe tip. A spin-polarized surface state of Co(0001) was found to exist at -0.43 eV relative to E(F), with FWHM of 0.23 eV in the spectra. The state exhibits negative magnetoresistance with an effective spin polarization of less than -23%, suggesting negatively high spin polarization of the surface state. Our first-principles calculations have supported the existence of the surface state. From the calculation, the state is identified as a minority spin Gamma-centered d(2)(z)-like surface state. PMID- 11863839 TI - One-dimensional energy dispersion of single-walled carbon nanotubes by resonant electron scattering. AB - We characterized the energy band dispersion near the Fermi level in single-walled carbon nanotubes using low-temperature scanning tunneling microscopy. Analysis of energy-dependent standing wave oscillations, which result from quantum interference of electrons resonantly scattered by defects, yields a linear energy dispersion near E(F), and indicates the importance of parity in scattering for armchair single-walled carbon nanotubes. Additionally, these data provide values of the tight-binding overlap integral and Fermi wave vector, in good agreement with previous work, but indicate that the electron coherence length is substantially shortened. PMID- 11863840 TI - Role of bulk and surface phonons in the decay of metal surface States. AB - We present a comprehensive theoretical investigation of the electron-phonon contribution to the lifetime broadening of the surface states on Cu(111) and Ag(111), in comparison with high-resolution photoemission results. The calculations, including electron and phonon states of the bulk and the surface, resolve the relative importance of the Rayleigh mode, being dominant for the lifetime at small hole binding energies. Including the electron-electron interaction, the theoretical results are in excellent agreement with the measured binding energy and temperature dependent lifetime broadening. PMID- 11863841 TI - Detection of spin-polarized electrons injected into a two-dimensional electron gas. AB - The spin-dependent mean-free path of electrons in a high-mobility InAs two dimensional electron gas (2DEG) is measured. Ferromagnetic metal/insulator/2DEG junctions are fabricated on a common channel in a nonlocal geometry and used as spin injectors and detectors. For electrons in spin-orbit eigenstates at 4.5 K, lower bounds for the spin mean-free path and relaxation time are Lambda(S) > or = 4.6 microm and tau(s) > or = 3.8 ps, respectively. The temperature dependence is weak over the range 4.5 4f(N-1)5d excitation spectra of Tb3+, Er3+, and Tm3+ in CaF2 and LiYF4. AB - High-energy transitions in the 4f(N) -->4f(N-1)5d excitation spectra of lanthanide ions in host crystals are usually broadened due to the short excited state lifetimes, whereas low-energy transitions, with longer excited-state lifetimes, may show fine structure. We report the surprising observation that for some materials fine structure is observed not only for the low-energy excitation bands but also for some high-energy transitions. The excited states that display fine structure are those for which the 5d electron is in the lowest crystal-field level but the 4f(N-1) core is in a highly excited state, indicating that the broadening depends only on the energy of the 5d electron and not on the total energy of the 4f(N-1)5d excited state. PMID- 11863853 TI - Quantum computation with trapped polar molecules. AB - We propose a novel physical realization of a quantum computer. The qubits are electric dipole moments of ultracold diatomic molecules, oriented along or against an external electric field. Individual molecules are held in a 1D trap array, with an electric field gradient allowing spectroscopic addressing of each site. Bits are coupled via the electric dipole-dipole interaction. Using technologies similar to those already demonstrated, this design can plausibly lead to a quantum computer with greater, approximately > or = 10(4) qubits, which can perform approximately 10(5) CNOT gates in the anticipated decoherence time of approximately 5 s. PMID- 11863854 TI - Model for bone strength and osteoporotic fractures. AB - Inner porous regions play a critical role in the load bearing capability of large bones. We show that an extension of disordered elastic networks [Chung et al., Phys. Rev. B 54, 15 094 (1996)] exhibits analogs of several known mechanical features of bone. The "stress backbones" and histograms of stress distributions for healthy and weak networks are shown to be qualitatively different. A hereto untested relationship between bone density and bone strength is presented. PMID- 11863855 TI - Optimal intracellular calcium signaling. AB - In many cell types, calcium is released from internal stores through calcium release channels upon external stimulation (e.g., pressure or receptor binding). These channels are clustered with a typical cluster size of about 20 channels, generating stochastic calcium puffs. We find that the clustering of the release channels in small clusters increases the sensitivity of the calcium response, allowing for coherent calcium responses at signals to which homogeneously distributed channels would not respond. PMID- 11863856 TI - Tank treading and unbinding of deformable vesicles in shear flow: determination of the lift force. AB - Deformation and tank-treading motion of flaccid vesicles in a linear shear flow close to a wall are quantitatively studied by light microscopy. Velocities of bounded vesicles obey Goldman's law established for rigid spheres. A progressive tilt and a transition of unbinding of vesicles are evidenced upon increasing the shear rate, gamma;. These observations disclose the existence of a viscous lift force, F(l), depending on the viscosity eta of the fluid, the radius R of the vesicle, its distance h from the substrate, and a monotonous decreasing function f(1-v) of the reduced volume v, in the following manner: F(l) = eta(gamma)(R(3)/h)f(1-v). This relation is valid for vesicles both close to and farther from the substrate. PMID- 11863857 TI - Coevolution of quasispecies: B-cell mutation rates maximize viral error catastrophes. AB - Coevolution of two coupled quasispecies is studied, motivated by the competition between viral evolution and adapting immune response. In this coadaptive model, besides the classical error catastrophe for high virus mutation rates, a second "adaptation" catastrophe occurs, when virus mutation rates are too small to escape immune attack. Maximizing both regimes of viral error catastrophes is a possible strategy for an optimal immune response, reducing the range of allowed viral mutation rates to a minimum. From this requirement, one obtains constraints on B-cell mutation rates and receptor lengths, yielding an estimate of somatic hypermutation rates in the germinal center in accordance with observation. PMID- 11863858 TI - Global optimization by energy landscape paving. AB - We introduce a novel heuristic global optimization method, energy landscape paving (ELP), which combines core ideas from energy surface deformation and tabu search. In appropriate limits, ELP reduces to existing techniques. The approach is very general and flexible and is illustrated here on two protein folding problems. For these examples, the technique gives faster convergence to the global minimum than previous approaches. PMID- 11863859 TI - Sequence determination from overlapping fragments: a simple model of whole-genome shotgun sequencing. AB - Assembling fragments randomly sampled from along a sequence is the basis of whole genome shotgun sequencing, a technique used to map the DNA of the human and other genomes. We calculate the probability that a random sequence can be recovered from a collection of overlapping fragments. We provide an exact solution for an infinite alphabet and in the case of constant overlaps. For the general problem we apply two assembly strategies and give the probability that the assembly puzzle can be solved in the limit of infinitely many fragments. PMID- 11863860 TI - Scaling theory for migration-driven aggregate growth. AB - We give a comprehensive description for the irreversible growth of aggregates by migration from small to large aggregates. For a homogeneous rate K(i;j) at which monomers migrate from aggregates of size i to those of size j, that is, K(ai;aj)similar to a(lambda)K(i;j), the mean aggregate size grows with time as t(1/(2-lambda)) for lambda<2. The aggregate size distribution exhibits distinct regimes of behavior that are controlled by the scaling properties of the migration rate from the smallest to the largest aggregates. Our theory applies to diverse phenomena such as the distribution of city populations, late stage coarsening of nonsymmetric binary systems, and models for wealth exchange. PMID- 11863862 TI - Comment on "elastoplastic contact between randomly rough surfaces". PMID- 11863861 TI - Waiting-time statistics of self-organized-criticality systems. AB - It is argued that a system governed by self-organized-criticality (SOC) dynamics can lack Poisson waiting-time statistics not only when the experimental resolution lies within the self-similar scale range but also if the system is slowly driven in a correlated way. This result thus suggests that waiting time statistics cannot be used as a necessary test for SOC behavior in real physical systems. PMID- 11863864 TI - Comment on "reetrant behavior of relaxation time with viscosity at varying composition in binary mixtures". PMID- 11863865 TI - Comment on "chaotic domain patterns in periodic inhomogeneous magnetic films". PMID- 11863867 TI - Misfolded loops decrease the effective rate of DNA hairpin formation. PMID- 11863869 TI - Dynamical instability of a condensate induced by a rotating thermal gas. AB - We study surface modes of the condensate in the presence of a rotating thermal cloud in an axisymmetric trap. By considering collisions that transfer atoms between the condensate and the noncondensate, we find that m>0 modes, which rotate in the same sense as the thermal cloud, damp less strongly than m<0 modes, where m is the polarity of the excitation. We show that above a critical angular rotation frequency, equivalent to the Landau stability criterion, m>0 modes become dynamically unstable, leading to the possibility of vortex nucleation. We also generalize our stability analysis to treat the case where the stationary state of the condensate already possesses a single vortex. PMID- 11863870 TI - Teleportation of a vacuum--one-photon qubit. AB - We report the experimental realization of teleporting a one-particle entangled qubit. The qubit is physically implemented by a two-dimensional subspace of states of a mode of the electromagnetic field, specifically, the space spanned by the vacuum and the one-photon state. Our experiment follows the line suggested by Lee and Kim [Phys. Rev. A 63, 012305 (2000)] and Knill, Laflamme, and Milburn [Nature (London) 409, 46 (2001)]. An unprecedented large value of the teleportation "fidelity" has been attained: F = (95.3 +/- 0.6)%. PMID- 11863871 TI - Observation of anomalous spin-state segregation in a trapped ultracold vapor. AB - We observe counterintuitive spin segregation in an inhomogeneous sample of ultracold, noncondensed rubidium atoms in a magnetic trap. We use spatially selective microwave spectroscopy to verify a model that accounts for the differential forces on two internal spin states. In any simple understanding of the cloud dynamics, the forces are far too small to account for the dramatic transient spin polarizations observed. The underlying mechanism remains to be elucidated. PMID- 11863872 TI - Stationary source of nonclassical or entangled atoms. AB - A scheme for generating continuous beams of atoms in nonclassical or entangled quantum states is proposed and analyzed. For this the recently suggested transfer technique of quantum states from light fields to collective atomic excitation by stimulated Raman adiabatic passage [M. Fleischhauer and M. D. Lukin, Phys. Rev. Lett. 84, 5094 (2000)] is employed and extended to matter waves. PMID- 11863873 TI - Consequence of superfluidity on the expansion of a rotating Bose-Einstein condensate. AB - We propose an easily detectable signature of superfluidity in rotating, vortex free gaseous Bose-Einstein condensates. We have studied the time evolution of the expansion of such a condensate after it is released from the confining trap. We find that if such a condensate is not initially rotating, then at some moment it will instantaneously achieve a circular cross section. If the condensate is initially rotating its irrotational flow and the conservation of angular momentum prevent the released condensate from attaining a circular cross section, since the instantaneous moment of inertia is then proportional to the asymmetry of this cross section. PMID- 11863874 TI - Direct observation of irrotational flow and evidence of superfluidity in a rotating Bose-Einstein condensate. AB - We have observed the expansion of vortex-free, rotating Bose condensates after their sudden release from a slowly rotating anisotropic trap. Conservation of angular momentum, combined with the constraint of irrotational flow, cause the rotating condensate to expand in a distinctively different way to one released from a static (nonrotating) trap. This difference provides clear experimental evidence of the purely irrotational velocity field associated with a superfluid. We observed this behavior in absorption images taken along the rotation axis. PMID- 11863875 TI - Phase fluctuations in atomic Bose gases. AB - We improve on the Popov theory for partially Bose-Einstein condensed atomic gases by treating the phase fluctuations exactly. As a result, the theory becomes valid in arbitrary dimensions and is able to describe the low-temperature crossover between three-, two-, and one-dimensional Bose gases, which is currently being explored experimentally. We consider both homogeneous and trapped Bose gases. PMID- 11863876 TI - Phonon spectrum and dynamical stability of a dilute quantum degenerate Bose-Fermi mixture. AB - We calculate the phonon excitation spectrum in a zero-temperature dilute boson fermion gaseous mixture. We show how the sound velocity changes due to the boson fermion interaction, and we determine the dynamical stability regime of a homogeneous mixture. We identify a resonant phonon-exchange interaction between the fermions as the physical mechanism leading to the instability. PMID- 11863877 TI - Formation and decay of vortex lattices in Bose-Einstein condensates at finite temperatures. AB - The dynamics of vortex lattices in stirred Bose-Einstein condensates have been studied at finite temperatures. The decay of the vortex lattice was observed nondestructively by monitoring the centrifugal distortions of the rotating condensate. The formation of the vortex lattice could be deduced from the increasing contrast of the vortex cores observed in ballistic expansion. In contrast to the decay, the formation of the vortex lattice is insensitive to temperature change. PMID- 11863878 TI - Series expansion calculation of persistence exponents. AB - We consider an arbitrary Gaussian stationary process X(T) with known correlator C(T), sampled at discrete times Tn = nDeltaT. The probability that (n+1) consecutive values of X have the same sign decays as Pn approximately exp( theta(D)Tn). We calculate the discrete persistence exponent theta(D) as a series expansion in the correlator C(DeltaT) up to fourteenth order, and extrapolate to DeltaT = 0 using constrained Pade approximants to obtain the continuum persistence exponent thetas. For the diffusion equation our results are in exceptionally good agreement with recent numerical estimates. PMID- 11863879 TI - Stringent neutron-star limits on large extra dimensions. AB - Supernovae (SNe) are copious sources for Kaluza-Klein (KK) gravitons which are generic for theories with large extra dimensions. These massive particles are produced with average velocities approximately 0.5c so that many of them are gravitationally retained by the SN core. Every neutron star thus has a halo of KK gravitons which decay into nu(nu), e(+)e(-), and gammagamma on time scales approximately 10(9) years. The EGRET gamma-flux limits (E(gamma) approximately 100 MeV) for nearby neutron stars constrain the compactification scale for n = 2 extra dimensions to M > or = 500 TeV, and M > or = 30 TeV for n = 3. The requirement that neutron stars are not excessively heated by KK decays implies M > or = 1700 TeV for n = 2, and M > or = 60 TeV for n = 3. PMID- 11863880 TI - Hard noncommutative loops resummation. AB - The noncommutative version of the Euclidean g2phi4 theory is considered. By using Wilsonian flow equations the ultraviolet renormalizability can be proved to all orders in perturbation theory. On the other hand, the infrared sector cannot be treated perturbatively and requires a resummation of the leading divergences in the two-point function. This is analogous to what is done in the hard thermal loops resummation of finite temperature field theory. Next-to-leading order corrections to the self-energy are computed, resulting in O(g3) contributions in the massless case, and O(g6logg2) in the massive one. PMID- 11863881 TI - Standard model on a D-brane. AB - We present a consistent string theory model which produces a simple extension of the standard model, consisting of a D3-brane at a simple orbifold singularity. We envision this as a local singularity within a warped compactification. The phenomenology of the model has some novel features. We note that, for the model to be viable, the scale of stringy physics must be in the multi-TeV range. There are natural hierarchies in the fermion spectrum and there are several possible experimental signatures of the model. PMID- 11863883 TI - Hadronic light-by-light scattering contribution to the muon g - 2: an effective field theory approach. AB - The hadronic light-by-light contribution to a(mu), the anomalous magnetic moment of the muon, is discussed from the point of view of an effective low-energy theory. As an application, the coefficient of the leading logarithm arising from the two-loop graphs involving two anomalous vertices is computed, and found to be positive. This corresponds to a positive sign for the pion-pole contribution to the hadronic light-by-light correction to a(mu), and to a sizable reduction of the discrepancy between the present experimental value of a(mu) and its theoretical counterpart in the standard model. PMID- 11863884 TI - Pion pole contribution to hadronic light-by-light scattering and muon anomalous magnetic moment. AB - We derive an analytic result for the pion pole contribution to the light-by-light scattering correction to the anomalous magnetic moment of the muon, a(mu) = (g(mu)-2)/2. Using the vector meson dominance model for the pion transition form factor, we obtain a(LBL,pi0)mu = +56x10(-11). PMID- 11863885 TI - Charged current universality in the minimal supersymmetric standard model. AB - We compute the complete one-loop contributions to low-energy charged current weak interaction observables in the minimal supersymmetric standard model (MSSM). We obtain the constraints on the MSSM parameter space which arise when precision low energy charged current data are analyzed in tandem with measurements of the muon anomaly. While the data allow the presence of at least one light neutralino, they also imply a pattern of mass splittings among first and second generation sleptons and squarks which contradicts predictions of widely used models for supersymmetry-breaking mediation. PMID- 11863886 TI - New limit on the electron electric dipole moment. AB - We present the result of our most recent search for T violation in 205Tl, which is interpreted in terms of an electric dipole moment of the electron d(e). We find d(e) = (6.9 +/- 7.4)x10(-28)e cm, which yields an upper limit /d(e)/ < or = 1.6x10(-27)e cm with 90% confidence. The present apparatus is a major upgrade of the atomic beam magnetic-resonance device used to set the previous limit on d(e). PMID- 11863888 TI - Kaon interferometry: a sensitive probe of the QCD equation of state? AB - We calculate the kaon-interferometry radius parameters for high-energy heavy-ion collisions, assuming a first-order phase transition from a thermalized quark gluon plasma (QGP) to a gas of hadrons. At high transverse momenta K(T) approximately 1 GeV/c direct emission from the phase boundary becomes important; the emission duration signal, i.e., the R(out)/R(side) ratio, and its sensitivity to T(c) (and thus to the latent heat) are enlarged. The QGP+hadronic rescattering transport model calculations do not yield unusually large radii (R(i) < or = 9 fm). Finite-momentum-resolution effects have a strong impact on the extracted interferometry parameters ( R(i) and lambda), as well as on the ratio R(out)/R(side). PMID- 11863889 TI - Elastic alpha-12C scattering and the 12C(alpha,gamma)16O E2 S factor. AB - Angular distributions of 12C(alpha,alpha)12C have been measured for E(alpha) = 2.6-8.2 MeV, at angles from 24 to 166, yielding 12 864 data points. R-matrix analysis of the ratios of elastic scattering yields a reduced width amplitude of gamma12 = 0.47 +/- 0.06 MeV(1/2) for the Ex = 6.917 MeV (2+) state in 16O(a = 5.5 fm). The dependence of the chi2 surface on the interaction radius a has been investigated and a deep minimum is found at a = 5.42(+0.16)(-0.27) fm. Using this value of gamma12, radiative alpha capture and 16N beta-delayed alpha-decay data, the S factor is calculated at E(c.m.) = 300 keV to be S(E2)(300) = 53(+13)(-18) keV b for destructive interference between the subthreshold resonance tail and the ground state E2 direct capture. PMID- 11863890 TI - Spectroscopy with random and displaced random ensembles. AB - Because of the time reversal invariance of the angular momentum operator J2, the average energies and variances at fixed J for random two-body Hamiltonians exhibit odd-even- J staggering that may be especially strong for J = 0. It is shown that upon ensemble averaging over random runs, this behavior is reflected in the yrast states. Displaced (attractive) random ensembles lead to rotational spectra with strongly enhanced B(E2) transitions for a certain class of model spaces. It is explained how to generalize these results to other forms of collectivity. PMID- 11863891 TI - Calculation of P, T-odd effects in 205TlF including electron correlation. AB - A method and codes for two-step correlation calculations of heavy-atom molecules have been developed, employing the generalized relativistic effective core potential (GRECP) and relativistic coupled cluster (RCC) methods at the first step, followed by nonvariational one-center restoration of proper four-component spinors in the heavy cores. Electron correlation is included for the first time in an ab initio calculation of the interaction of the permanent P,T-odd proton electric dipole moments with the internal electromagnetic field in a molecule. Inclusion of electron correlation by GRECP/RCC has a major effect on the P,T-odd parameters of 205TlF, decreasing M by 17% and X by 22%. PMID- 11863892 TI - Circular dichroism in K-shell ionization from fixed-in-space CO and N2 molecules. AB - We have measured the angular distributions of 1s photoelectrons excited by circularly and linearly polarized light from fixed-in-space CO and N2 molecules, in the vicinity of their shape resonances. A strong circular dichroism, i.e., a strong dependence on the sense of rotation of the polarization vector of the photons, is found for both molecules. State-of-the-art one-electron multiple scattering and partially correlated random phase approximation calculations are in good agreement with many, but not all, aspects of the experimental data. PMID- 11863893 TI - Measurement of the electron affinity of cerium. AB - The electron affinity of cerium has been measured using laser photodetachment electron spectroscopy. The electron affinity of Ce(1G4) was determined to be 0.955 +/- 0.026 eV. The data also show that Ce- has at least two bound excited states with binding energies of 0.921 +/- 0.025 eV and 0.819 +/- 0.027 eV relative to the (1G4) ground state of the cerium atom. The present experimental measurements are compared to recent calculations of the energy levels of Ce-. Strong disagreement with the most recent theoretical atomic structure calculations highlights the complicated nature of this particular lanthanide. PMID- 11863894 TI - Evidence for Fermi-shuttle ionization in intermediate velocity C+ + Xe collisions. AB - Experimental evidence has been found for consecutive projectile-target-projectile (triple) and projectile-target-projectile-target (quadruple) "ping-pong" scattering of ionized target electrons in single C+ +Xe collisions at 150 and 233 keV/u impact energies. Distinct signatures of the multiple electron scattering contributions to the high-energy part (300-3400 eV) of the double differential electron spectra have been separated and identified with the help of reference measurements using He+ projectile ions and different calculations. PMID- 11863895 TI - Ultraprecise measurement of optical frequency ratios. AB - We developed a novel technique for frequency measurement and synthesis, based on the operation of a femtosecond comb generator as transfer oscillator. The technique can be used to measure frequency ratios of any optical signals throughout the visible and near-infrared part of the spectrum. Relative uncertainties of 10(-18) for averaging times of 100 s are possible. Using a Nd:YAG laser in combination with a nonlinear crystal we measured the frequency ratio of the second harmonic nu(SH) at 532 nm to the fundamental nu(0) at 1064 nm, nu(SH)/nu(0) = 2.000 000 000 000 000 001x (1 +/- 7x10(-19)). PMID- 11863896 TI - Measurement of the superluminal group velocity of an ultrashort Bessel beam pulse. AB - The superluminal group velocity of an ultrashort optical Bessel beam pulse is measured over its entire depth of field, corresponding to approximately 2x10(4) optical wavelengths. The method used is to measure the traveling ionization front induced by the pulse. PMID- 11863882 TI - Study of B0 --> J/psiK(*)0pi(+)pi(-) decays with the collider detector at Fermilab. AB - We report a study of the decays B0 --> J/psiK(*)0pi(+)pi(-), which involve the creation of a uu or dd quark pair in addition to a b -->c(cs) decay. The data sample consists of 110 pb(-1) of pp collisions at square root[s] = 1.8 TeV collected by the CDF detector at the Fermilab Tevatron collider during 1992-1995. We measure the branching fractions to be B(B0 --> J/psiK(*0)pi(+)pi(-)) = (6.6 +/ 1.9 +/- 1.1)x10(-4) and B(B0 --> J/psiK0pi(+)pi(-)) = (10.3 +/- 3.3 +/- 1.5)x10( 4). Evidence is seen for contributions from psi(2S)K(*)0, J/psiK0rho(0), J/psiK(*+)pi(-), and J/psiK1(1270). PMID- 11863898 TI - Experimental evidence for soliton explosions. AB - We show, experimentally and numerically, that Ti:sapphire mode-locked lasers can operate in a regime in which they intermittently produce exploding solitons. This happens when the laser operates near a critical point. Explosions happen spontaneously, but external perturbations can trigger them. In stable operation, all explosions have similar features, but are not identical. The characteristics of the explosions depend on the intracavity dispersion. PMID- 11863897 TI - Stable spinning optical solitons in three dimensions. AB - We introduce spatiotemporal spinning solitons (vortex tori) of the three dimensional nonlinear Schrodinger equation with focusing cubic and defocusing quintic nonlinearities. The first ever found completely stable spatiotemporal vortex solitons are demonstrated. A general conclusion is that stable spinning solitons are possible as a result of competition between focusing and defocusing nonlinearities. PMID- 11863899 TI - Argon rectification and the cause of light emission in single-bubble sonoluminescence. AB - In single-bubble sonoluminescence, repeated brief flashes of light are produced in a gas bubble strongly driven by a periodic acoustic field. A startling hypothesis has been made by Lohse and co-workers [Phys. Rev. Lett. 78, 1359 (1997)] that the non-noble gases in an air bubble undergo chemical reaction into soluble products, leaving only argon. In the present work, this dissociation hypothesis is supported by simulations, although the associated temperatures of about 7000 K seem too low for bremsstrahlung, which has been proposed as the dominant light emission mechanism. This suggests that emission from water vapor and its reaction products, heretofore not included, may play an important role. PMID- 11863900 TI - Boosting sonoluminescence with a high-intensity ultrasonic pulse focused on the bubble by an adaptive array. AB - Single-bubble sonoluminescence is characterized by a great concentration of energy during the collapse of a gas bubble, which leads to the generation of photons from low-frequency ultrasound. The narrow stability domain of sonoluminescence has limited previous attempts to reinforce this inertial confinement in order to generate photons of higher energy or to ignite a nuclear fusion reaction. We present a new experimental approach where an ultrasonic pulse of high frequency is adaptively focused on the bubble during the collapse. Using an array of eight transmitters, a pressure pulse of 0.7 MPa doubles the flash intensity; this technique can easily be extended to higher pressure. PMID- 11863901 TI - Scaling at the selective withdrawal transition through a tube suspended above the fluid surface. AB - In the selective withdrawal experiment fluid is withdrawn through a tube with its tip suspended a distance S above an unperturbed two-fluid interface. At low withdrawal rates, Q, the interface forms a steady state hump and only the upper fluid is withdrawn. When Q is increased (or S decreased), the interface undergoes a transition so that the lower fluid is entrained with the upper one, forming a thin steady-state spout. Near this discontinuous transition the hump curvature becomes very large and displays power-law scaling behavior. This scaling is used to show that steady-state profiles for humps at different flow rates and tube heights can all be scaled onto a single similarity profile. PMID- 11863887 TI - Search for new heavy particles in the WZ0 final state in pp collisions at square root[s] = 1.8 TeV. AB - We present the first general search for new heavy particles, X, which decay via X --> WZ0 --> e(nu)+jj as a function of M(X) and Gamma(X) in pp collisions at square root[s] = 1.8 TeV. No evidence is found for production of X in 110 pb(-1) of data collected by the Collider Detector at Fermilab. General cross section limits are set at the 95% C.L. as a function of mass and width of the new particle. The results are further interpreted as mass limits on the production of new heavy charged vector bosons which decay via W' --> WZ0 in an extended gauge model as a function of the width, Gamma(W'), and mixing factor between the W' and the standard model W bosons. PMID- 11863902 TI - Lasetron: a proposed source of powerful nuclear-time-scale electromagnetic bursts. AB - Electromagnetic bursts of substantial energy on a nuclear time scale of 10(-21) 10(-22) s [zeptosecond (zs) to sub-zs] can theoretically be generated by a perawatt or multiterawatt laser beam focused on a subwavelength-size solid particle or thin wire. Terawatt laser in a similar setup could be instrumental in reaching the subattosecond domain. The system may also generate a half-cycle pulse magnetic field on astrophysical scale up to approximately 10(6) T. PMID- 11863903 TI - Measurements of harmonic wake fields excited by rough surfaces. AB - An experiment has been carried out at the TESLA Test Facility linac to investigate the wake fields generated by picosecond electron bunches in narrow beam pipes with an artificially roughened inner surface. The energy structure imposed on the bunches by the wake fields has been analyzed with a magnetic spectrometer. Strong harmonic-wake-field effects are observed as expected from simulations in which the rough surface is modeled by a dielectric layer. PMID- 11863904 TI - Reduction of the ion drive and rho*(theta) scaling of the neoclassical tearing mode. AB - In the theoretical description of the neoclassical tearing mode it is usually assumed that the ion banana width w(b) is much smaller than the island width W. This assumption is questionable at least for the island size at the mode onset. We show that a significant fraction of the (ion) bootstrap current survives inside the island when w(b) is comparable to W. This effect also leads to a linear scaling of beta(onset)theta with the normalized ion poloidal gyroradius rho*(theta), in agreement with the experimental results of ASDEX Upgrade. PMID- 11863905 TI - Exponentially growing tearing modes in Rijnhuizen Tokamak Project plasmas. AB - The local measurement of the island width w, around the resonant surface, allowed a direct test of the extended Rutherford model [P. H. Rutherford, PPPL Report 2277 (1985)], describing the evolution of radiation-induced tearing modes prior to disruptions of tokamak plasmas. It is found that this model accounts very well for the observed exponential growth and supports radiation losses as being the main driving mechanism. The model implies that the effective perpendicular electron heat conductivity in the island is smaller than the global one. Comparison of the local measurements of w with the magnetic perturbed field B showed that w proportional to B1/2 was valid for widths up to 18% of the minor radius. PMID- 11863906 TI - Direct coupling of magnetic fields to tunneling systems in glasses. AB - We report on investigations of spontaneous polarization echoes in the nonmagnetic multicomponent glass BaO-Al2O3-SiO2 in static magnetic fields. While the echo decay is only marginally influenced, the echo amplitude depends strongly on magnetic fields. It seems that the intrinsic magnetic moment of tunneling systems causes dephasing effects which are detected in our echo experiments. In addition we find a strong increase of the echo amplitude with magnetic fields. This result shows that the coupling of the tunneling systems to magnetic fields is surprisingly strong and cannot be understood on the basis of current theories. PMID- 11863907 TI - Persistent tunneling currents and magnetic-field effects in glasses. AB - Recent experiments on insulating glasses showed an unexpected variation of the dielectric constant with a static magnetic field and, in particular, oscillations with an apparent flux quantum of about 10(-5)h/e. We propose an explanation for these phenomena in terms of a model based on pairs of two-level systems. A weak interaction is shown to give rise to quantum states with a net circular current and a linear field dependence of the energy levels. Under experimental conditions, the dynamic susceptibility shows oscillatory behavior, with an effective flux quantum of the order of 10(-5)h/e. PMID- 11863908 TI - Evidence of kinetic trapping in clusters of C60 molecules. AB - Molecular dynamics simulations of the growth of nanoclusters of C60 provide convincing evidence that experimental magic numbers, which are associated with structures based on Mackay icosahedra, are of kinetic origin. This finding resolves a long-standing contradiction between the experimental observations and the theoretically predicted most stable structures. Our results show that, even if a sticky intermolecular potential energetically disfavors icosahedral structures, the latter are frequently produced because the stickiness of the potential itself enhances kinetic trapping phenomena. PMID- 11863909 TI - Lattice dynamics of solid xenon under pressure. AB - We use density-functional perturbation theory to obtain the phonon spectrum of fcc xenon under pressure. Thermodynamic properties obtained within the quasiharmonic approximation are in fair to good agreement with experiment at zero pressure. The transition pressure from the fcc to hcp phase is predicted to occur at 5 GPa. The fcc structure is found to be dynamically stable up to a pressure of 100 GPa, beyond which the phonon modes at the X and L symmetry points soften. We attribute the observed sluggish kinetics of the fcc-hcp transition to the small energy difference between the phases as well as to the high dynamical stability of the fcc phase. PMID- 11863910 TI - Bond-stretching-phonon anomalies in stripe-ordered La1.69Sr0.31NiO4. AB - We report a neutron scattering study of bond-stretching phonons in La1.69Sr0.31NiO4, a doped antiferromagnet in which the added holes order in diagonal stripes at 45 to the Ni-O bonds. For the highest-energy longitudinal optical mode along the bonds, a softening of 20% is observed between the Brillouin zone center and the zone boundary. At 45 to the bonds, a splitting of the same magnitude is found across much of the zone. Surprisingly, the charge ordering wave vector plays no apparent role in the anomalous dispersions. The implications for related anomalies in the cuprates are discussed. PMID- 11863911 TI - Ab initio investigations of lithium diffusion in carbon nanotube systems. AB - Li-nanotube systems can substantially improve the capacity of Li-ion batteries by utilizing both nanotube exteriors and interiors. Our ab initio simulations show that while Li motion through the sidewalls is forbidden, Li ions can enter tubes through topological defects containing at least nine-sided rings, or through the ends of open-ended nanotubes. Once inside, their motion is not diffusion limited. These results suggest that "damaging" nanotube ropes by either chemical or mechanical means will yield superior material for electrochemical storage. PMID- 11863912 TI - Random packings of frictionless particles. AB - We conduct numerical simulations of random packings of frictionless particles at T = 0. The packing fraction where the pressure becomes nonzero is the same as the jamming threshold, where the static shear modulus becomes nonzero. The distribution of threshold packing fractions narrows, and its peak approaches random close packing as the system size increases. For packing fractions within the peak, there is no self-averaging, leading to exponential decay of the interparticle force distribution. PMID- 11863913 TI - Localized metallic conductivity and self-healing during thermal reduction of SrTiO3. AB - The occurrence of metallic conductivity in SrTiO3 single crystals is reported for reduction under low partial pressure of oxygen at 800 degrees C. This transition is shown to result from the formation of a high concentration of vacancy defects along a network of extended defects within the skin region. A self-healing phenomenon is observed for progressive reduction which causes the concentration of initially introduced defects to decrease in the course of heat treatment and leads to a breakdown of the metallic conductivity as well as a substantial loss of optical subgap absorption. PMID- 11863914 TI - Self-healing slip pulses along a gel/glass interface. AB - We present experimental evidence of self-healing shear cracks at a gel/glass interface. This system exhibits two dynamical regimes depending on the driving velocity: steady sliding at high velocity (>V(c) approximately 100--125 microm/s), characterized by a shear-thinning rheology, and periodic stick-slip dynamics at low velocity. In this last regime, slip occurs by propagation of pulses that restick via a "healing instability" occurring when the local sliding velocity reaches the macroscopic transition velocity V(c). At driving velocities close below V(c), the system exhibits complex spatiotemporal behavior. PMID- 11863915 TI - Crumpling a thin sheet. AB - Crumpled sheets have a surprisingly large resistance to further compression. We have studied the crumpling of thin sheets of Mylar under different loading conditions. When placed under a fixed compressive force, the size of a crumpled material decreases logarithmically in time for periods up to three weeks. We also find hysteretic behavior when measuring the compression as a function of applied force. By using a pretreating protocol, we control this hysteresis and find reproducible scaling behavior for the size of the crumpled material as a function of the applied force. PMID- 11863916 TI - Photon-induced thermal desorption of CO from small metal-carbonyl clusters. AB - Thermal CO desorption from photoexcited free metal-carbonyl clusters has been resolved in real time using two-color pump-probe photoelectron spectroscopy. Sequential energy dissipation steps between the initial photoexcitation and the final desorption event, e.g., electron relaxation and thermalization, have been resolved for Au2(CO)(-) and Pt2(CO)5-. The desorption rates for the two clusters differ considerably due to the different numbers of vibrational degrees of freedom. The unimolecular CO-desorption thresholds of Au2(CO)(-) and Pt2(CO)5- have been approximated by means of a statistical Rice-Ramsperger-Kassel calculation using the experimentally derived desorption rate constants. PMID- 11863917 TI - Hydrodynamic force measurements: boundary slip of water on hydrophilic surfaces and electrokinetic effects. AB - The hydrodynamic drainage force of aqueous medium between smooth hydrophilic surfaces was measured with the colloidal probe technique up to shear rates of typically 10(4) s(-1). Measured force curves were compared to simulations. To reach agreement between experimental and simulated force curves, the hydrodynamic force had to be fitted with a model allowing for boundary slippage. Boundary slip was characterized by a slip length of 8-9 nm. Force measurements with charged surfaces could be simulated taking only hydrodynamic and electrostatic double layer forces into account. PMID- 11863918 TI - Observation of solitary elastic surface pulses. AB - The formation of solitary elastic surface pulses from laser-generated pulselike initial conditions is reported. The nonlinearity of the medium is compensated by both normal dispersion and anomalous dispersion, which were realized by coating isotropic fused silica by a metal and titanium nitride film, respectively. As an anisotropic material, silicon covered with an oxide layer was studied. The experimental results agree with numerical simulations carried out with a nonlocal evolution equation, which describes nonlinear propagation of surface acoustic waves in a dispersive medium. PMID- 11863919 TI - Origin of anomalously long interatomic distances in suspended gold chains. AB - The discovery of long bonds in gold atom chains has represented a challenge for physical interpretation. In fact, interatomic distances frequently attain 3.0-3.6 A values, and distances as large as 5.0 A may be occasionally observed. Here we studied gold chains by transmission electron microscopy and performed theoretical calculations using cluster ab initio density functional formalism. We show that the insertion of two carbon atoms is required to account for the longest bonds, while distances above 3 A may be due to a mixture of clean and one C atom contaminated bonds. PMID- 11863920 TI - Nitridation of Si(100)-( 2x1) surface by NH3: a quantum chemical cluster model study. AB - Based on density functional cluster model calculations, we present the first detailed mechanisms for the complete decomposition of NH3 to NHx(a) (x = 0-2) on the Si(100)-(2x1) surface. Three kinds of elementary processes, namely, N-H bond cleavage, NHx(a) insertion into the Si-Si surface dimer bond or backbond, and H2 libration, are investigated. A plausible microscopic mechanism for the nitridation of Si(100)-(2x1) surface by NH3 is proposed. PMID- 11863921 TI - Stereochemistry on Si(001): angular dependence of H2 dissociation. AB - The angular dependence of the dissociative adsorption of molecular hydrogen at terrace and step sites of vicinal single-domain Si(001) surfaces was investigated by means of molecular beam techniques and optical second-harmonic generation. A strongly anisotropic behavior was observed for terrace adsorption with polar distributions of cos3theta and cos12theta parallel and perpendicular to the dimer, respectively. The D(B)-steps show enhanced reactivity under glancing incidence in the upwards direction. The results are traced back to the directionality of the covalent surface bonds. PMID- 11863922 TI - Weak anisotropy and disorder dependence of the in-plane magnetoresistance in high mobility (100) Si-inversion layers. AB - We report studies of the magnetoresistance (MR) in a two-dimensional electron system in (100) Si-inversion layers, for perpendicular and parallel orientations of the current with respect to the magnetic field in the 2D plane. The magnetoresistance is almost isotropic; this result does not support the suggestion of its orbital origin. In the hopping regime, however, the MR contains a weak anisotropic component that is nonmonotonic in the magnetic field. We found that the field, at which the MR saturates, varies for different samples by a factor of 2 at a given carrier density. Therefore, the saturation of the MR cannot be identified with the complete spin polarization of free carriers. PMID- 11863923 TI - Thermodynamic investigation of the charge-ordered insulating state of the quasi one-dimensional organic system (DI-DCNQI)2Ag. AB - Heat capacity measurements of a charge-ordered organic conductor (DI-DCNQI)2Ag have been performed in a temperature range between 0.3 and 14 K. We found no thermal anomaly at the Neel temperature ( T(N) = 5.5 K) but instead a T-linear term suggestive of the spin excitations of one-dimensional character in the charge-ordered insulating state. The analysis of the T-linear term and the excess entropy indicates that the charge fluctuations in the charge-ordered state influence the growth of spin excitations at elevated temperatures, which seems to be a peculiar aspect of a 1D charge-ordered system. PMID- 11863924 TI - Microscopic model of superconductivity in carbon nanotubes. AB - We propose the model of a manifold of one-dimensional interacting electron systems to account for the superconductivity observed in ropes of nanotubes. We rely on the strong suppression of single-particle hopping between neighboring nanotubes in a disordered rope and conclude that the tunneling takes place in pairs of electrons, which are formed within each nanotube due to the existence of large superconducting correlations. Our estimate of the transition temperature is consistent with the values that have been measured experimentally in ropes with about 100 metallic nanotubes. PMID- 11863925 TI - Transmission phase of a quantum dot with Kondo correlation near the unitary limit. AB - The complex transmission amplitude of a quantum dot (QD) with Kondo correlation was measured near the unitary limit. The transmission phase was observed to evolve almost linearly over a range of about 1.5 pi when the Fermi energy was scanned through a spin degenerate energy level of the QD. Surprisingly, the phase in the Coulomb Blockade regime, with one more electron entering the dot, was strongly affected by a preexistence of Kondo correlation. These results suggest that a full explanation of the Kondo effect may go beyond the framework of the Anderson model. PMID- 11863927 TI - Quasiparticle conductivities in disordered d-wave superconductors. AB - We study the quasiparticle transport coefficients in disordered d-wave superconductors. We find that spin and charge excitations are generally localized unless magnetic impurities are present. If the system is close to a nesting point in the impurity-scattering unitary limit, the tendency towards localization is reduced while the quasiparticle density of states gets enhanced by disorder. We also show that the residual repulsive interaction among quasiparticles has a delocalizing effect and increases the density of states. PMID- 11863928 TI - Insulating and fractional quantum hall states in the first excited Landau level. AB - The observation of new insulating phases of two-dimensional electrons in the first excited Landau level is reported. These states, which are manifested as reentrant integer quantized Hall effects, exist alongside well-developed even denominator fractional quantized Hall states at nu = 7/2 and 5/2 and new odd denominator states at nu = 3+1/5 and 3+4/5. PMID- 11863926 TI - Sensitivity to disorder of the metallic state in the ruthenates. AB - We report the results of transport measurements on SrRuO3, Sr3Ru2O7, and CaRuO3. In SrRuO3 and Sr3Ru2O7, our findings are consistent with the predictions of Fermi liquid theory, in contrast to previous reports based on samples with much shorter mean free paths. In CaRuO3, however, a T1.5 power law is seen in the resistivity in the high purity samples studied here. Our work gives concrete evidence that even the metallic state of the ruthenates is highly sensitive to disorder. PMID- 11863929 TI - Low-temperature electronic heat transport in La2-xSrxCuO4 single crystals: unusual low-energy physics in the normal and superconducting states. AB - The thermal conductivity kappa is measured in a series of La2-xSrxCuO4 (0 < or = x < or = 0.22) single crystals down to 90 mK to elucidate the evolution of the residual electronic thermal conductivity kappa(res), which probes the extended quasiparticle states in the d-wave gap. We found that kappa(res)/T grows smoothly, except for a 1/8 anomaly, above x approximately 0.05, and shows no discontinuity at optimum doping, indicating that the behavior of kappa(res)/T is not governed by the metal-insulator crossover in the normal state; as a result, kappa(res)/T is much larger than what the normal-state resistivity would suggest in the underdoped region, which highlights the peculiarities in the low-energy physics in the cuprates. PMID- 11863930 TI - Suppression of the "Quasiclassical" proximity gap in correlated-metal- superconductor structures. AB - We study the energy and spatial dependence of the local density of states in a superconductor--correlated-metal--superconductor Josephson junction, where the correlated metal is a non-Fermi liquid (described by the Falicov-Kimball model). Many-body correlations are treated with dynamical mean-field theory, extended to inhomogeneous systems. While quasiclassical theories predict a minigap in the spectrum of a disordered Fermi liquid which is proximity-coupled within a mesoscopic junction, we find that increasing electron correlations destroy any minigap that might be opened in the absence of many-body correlations. PMID- 11863931 TI - Delocalized quasiparticles in the vortex state of an overdoped high-T(c) superconductor probed by 63Cu NMR. AB - We report the spin Knight shift (K(s)) and the nuclear spin-lattice relaxation rate (1/T1) in the vortex state as a function of magnetic field (H) up to 28 T in the high-Tc superconductor TlSr2CaCu2O6.8 (Tc = 68 K). At low temperatures well below Tc, both K(s) and 1/T1 measured around the middle point between the two nearest vortices (saddle point) increase substantially with increasing field, which indicate that the quasiparticle states with an ungapped spectrum are extended outside the vortex cores in a d-wave superconductor. The density of states (DOS) around the saddle point is found to be kappaN(0)square root[H/H(c2)], with kappa = 0.5-0.7 and N0 being the normal-state DOS. PMID- 11863932 TI - Aspect-ratio scaling and the stiffness exponent theta for Ising spin glasses. AB - We introduce the technique of aspect-ratio scaling to study the scale dependence of interfacial energies in Ising spin glasses, and we show how one can use it to determine the stiffness exponent theta in a clean way, with results that are independent of the domain-wall-forcing boundary conditions imposed on the system. In space dimension d = 2 we obtain theta = -0.282(3) for a Gaussian distribution of exchange interactions. PMID- 11863933 TI - Chiral Kosterlitz-Thouless transition in the frustrated Heisenberg antiferromagnet on a pyrochlore slab. AB - Ordering of the geometrically frustrated two-dimensional Heisenberg antiferromagnet on a pyrochlore slab is studied by Monte Carlo simulations. In contrast to the kagome Heisenberg antiferromagnet, the model exhibits locally noncoplanar spin structures at low temperatures, bearing nontrivial chiral degrees of freedom. Under certain conditions, the model exhibits a novel Kosterlitz-Thouless-type transition at a finite temperature associated with these chiral degrees of freedom. PMID- 11863934 TI - Low-temperature transport in Heisenberg chains. AB - A technique to determine accurately transport properties of integrable and nonintegrable quantum-spin chains at finite temperatures by quantum Monte Carlo is presented. The reduction of the Drude weight by interactions in the integrable gapless regime is evaluated. Evidence for the absence of Drude weight in the gapless regime of a nonintegrable system with longer-ranged interactions is presented. We estimate the effect of the nonintegrability on the transport properties and compare with recent experiments on one-dimensional quantum-spin chains. PMID- 11863935 TI - First-order transition in the spin dynamics of geometrically frustrated Yb2Ti2O7. AB - Using neutron diffraction, 170Yb Mossbauer and muon spin relaxation spectroscopies, we have examined the pyrochlore Yb2Ti2O7, where the Yb3+S' = 1/2 ground state has planar anisotropy. Below approximately 0.24 K, the temperature of the known specific-heat lambda transition, there is no long range magnetic order. We show that the transition corresponds to a first-order change in the fluctuation rate of the Yb3+ spins. Above the transition temperature, the rate, in the GHz range, follows a thermal excitation law, whereas below, the rate, in the MHz range, is temperature independent, indicative of a quantum fluctuation regime. PMID- 11863936 TI - Temperature dependence of a single Kondo impurity. AB - Recent advances in scanning tunneling microscopy have allowed the observation of the Kondo effect for individual magnetic atoms. One hallmark of the Kondo effect is a strong temperature-induced broadening of the Kondo resonance. In order to test this prediction for individual impurities, we have investigated the temperature dependent electronic structure of isolated Ti atoms on Ag(100). We find that the Kondo resonance is strongly broadened in the temperature range T = 6.8 K to T = 49.0 K. These results are in good agreement with theoretical predictions for Kondo impurities in the Fermi liquid regime, and confirm the role of electron-electron scattering as the main thermal broadening mechanism. PMID- 11863937 TI - Quasielastic neutron scattering in the high-field phase of a Haldane antiferromagnet. AB - Inelastic neutron scattering experiments on the Haldane-gap quantum antiferromagnet Ni(C5D14N2)2-N3(PF6) are performed in magnetic fields below and above the critical field H(c) at which the gap closes. Quasielastic neutron scattering is found for H>H(c), indicating topological excitations in the high field phase. PMID- 11863938 TI - Electron correlation effects in resonant inelastic X-ray scattering of NaV2O5. AB - Element- and site-specific resonant inelastic x-ray scattering spectroscopy (RIXS) is employed to investigate electron correlation effects in NaV2O5. In contrast to single photon techniques, RIXS at the vanadium L3 edge is able to probe d-d* transitions between V d-bands. A sharp energy loss feature is observed at -1.56 eV, which is well reproduced by a model calculation including correlation effects. The calculation identifies the loss feature as excitation between the lower and upper Hubbard bands and permits an accurate determination of the Hubbard interaction term U = 3.0 +/- 0.2 eV. PMID- 11863939 TI - Drastic reduction of plasmon damping in gold nanorods. AB - The dephasing of particle plasmons is investigated using light-scattering spectroscopy on individual gold nanoparticles. We find a drastic reduction of the plasmon dephasing rate in nanorods as compared to small nanospheres due to a suppression of interband damping. The rods studied here also show very little radiation damping, due to their small volumes. These findings imply large local field enhancement factors and relatively high light-scattering efficiencies, making metal nanorods extremely interesting for optical applications. Comparison with theory shows that pure dephasing and interface damping give negligible contributions to the total plasmon dephasing rate. PMID- 11863940 TI - Real-time measurement of the orientational dynamics of aqueous solvation shells in bulk liquid water. AB - We present a study of the orientational dynamics of aqueous solvation shells of halogenic anions in bulk water solution with femtosecond two-color midinfrared spectroscopy. These orientational dynamics have time constants between 2.5 and 12 ps, depending on the type of anion and the temperature. We find that the solvation shell of the larger ion I- shows faster dynamics than that of the smaller ions Cl- and Br-. PMID- 11863941 TI - Optimizing a phase gate using quantum interference. AB - A controlled interference is proposed to reduce, by two orders of magnitude, the decoherence of a quantum gate for which the gate fidelity is limited by coupling to states other than the /0> and /1> qubit states. This phenomenon is demonstrated in an ultracold neutral atom implementation of a phase gate using qubits based on motional states in individual wells of an optical lattice. PMID- 11863942 TI - Maternal effects in molecular evolution. AB - We introduce a model of molecular evolution in which the fitness of an individual depends both on its own and on the parent's genotype. The model can be solved by means of a nonlinear mapping onto the standard quasispecies model. The dependency on the parental genotypes cancels from the mean fitness, but not from the individual sequence concentrations. For finite populations, the position of the error threshold is very sensitive to the influence from parent genotypes. In addition to biological applications, our model is important for understanding the dynamics of self-replicating computer programs. PMID- 11863943 TI - SO3 symmetry breaking mechanism for orientation and spatial frequency tuning in the visual cortex. AB - A dynamical model of orientation and spatial frequency tuning in a cortical hypercolumn is presented. The network topology is taken to be a sphere whose poles correspond to orientation pinwheels associated with high and low spatial frequency domains, respectively. Recurrent interactions within the sphere generate a tuned response via an SO3 symmetry breaking mechanism. PMID- 11863944 TI - Role of laser-induced plasma formation in pulsed cellular microsurgery and micromanipulation. AB - We investigate experimentally the physical processes underlying pulsed cellular microsurgery and micromanipulation using nanosecond 532- and 1064-nm laser pulses focused at high numerical aperture. We find that the laser parameters employed for many microirradiation techniques are congruent with those leading to optical breakdown in water. We determine the size and shape of the laser-induced plasma, pressure of the emitted shock wave, and size and energy of the cavitation bubble formed by the expanding plasma. We discuss implications of the results for biophysical microirradiation procedures. PMID- 11863946 TI - Comment on "Coulomb explosion patterns of fast C60 clusters in solids". PMID- 11863945 TI - Comment on "unique translation between Hamiltonian operators and functional integrals". PMID- 11863947 TI - Stable skyrmions in two-component Bose-Einstein condensates. AB - We show that stable Skyrmions exist in two-component atomic Bose-Einstein condensates, in the regime of phase separation. Using full three-dimensional simulations we find the stable Skyrmions with topological charges Q = 1 and 2, and compute their properties. With reference to these computations we suggest the salient features of an experimental setup in which they might realized. PMID- 11863948 TI - Growth of Bose-Einstein condensates from thermal vapor. AB - We report on a quantitative study of the growth process of (87)Rb Bose-Einstein condensates. By continuous evaporative cooling we directly control the thermal cloud from which the condensate grows. We compare the experimental data with the results of a theoretical model based on quantum kinetic theory. We find quantitative agreement with theory for the situation of strong cooling, whereas in the weak cooling regime a distinctly different behavior is found in the experiment. PMID- 11863949 TI - Evidence for nearby supernova explosions. AB - Supernova (SN) explosions are one of the most energetic---and potentially lethal- -phenomena in the Universe. We show that the Scorpius-Centaurus OB association, a group of young stars currently located at approximately 130 pc from the Sun, has generated 20 SN explosions during the last 11 Myr, some of them probably as close as 40 pc to our planet. The deposition on Earth of (60)Fe atoms produced by these explosions can explain the recent measurements of an excess of this isotope in deep ocean crust samples. We propose that approximately 2 Myr ago, one of the SNe exploded close enough to Earth to seriously damage the ozone layer, provoking or contributing to the Pliocene-Pleistocene boundary marine extinction. PMID- 11863950 TI - Unification and hierarchy from 5D anti-de Sitter space. AB - We show that perturbative high scale unification and a solution to the hierarchy problem are possible with extra dimensions in the context of the warped geometry of 5D anti-de Sitter space ( AdS(5)). This is possible because the couplings for bulk gauge bosons run logarithmically below the AdS(5) curvature scale. The calculation is done in five dimensions, rather than in the effective theory, which is strongly coupled above the TeV scale. PMID- 11863952 TI - Hypernuclear fine structure in (9)(lambda)Be. AB - With a germanium detector array (Hyperball), we observed two gamma-ray peaks corresponding to the two transitions (5/2(+)-->1/2(+) and 3/2(+)-->1/2(+)) in the (9)(Lambda)Be hypernucleus which was produced by the 9Be(K-,pi(-)) reaction. The energies of the gamma rays are 3029 +/- 2 +/- 1 keV and 3060 +/- 2 +/- 1 keV. The energy difference was measured to be 31.4(+2.5)(-3.6) keV, which indicates a very small Lambda-spin-dependent spin-orbit force between a Lambda and a nucleon. This is the smallest level splitting by far ever measured in a hypernucleus. PMID- 11863953 TI - Symmetry-resolved vibrational spectroscopy for the C 1s(-1)2 pi(u) Renner-Teller pair states in CO(2). AB - Symmetry-resolved excitation spectra have been measured for the Renner-Teller pair states A(1) and B(1) split from the core-excited C 1s(-1)2 pi(u) state in CO(2). A vibrational progression with the spacings of approximately 145 meV is found in both the A(1) and B(1) spectra at different energies and assigned to the symmetric stretching mode caused in the B(1) linear state, with the help of ab initio calculations. Appearance of the vibrations in the A(1) spectrum is interpreted as due to non-adiabatic coupling between the A(1) and B(1) states via the bending motion. PMID- 11863954 TI - Atom lithography with a holographic light mask. AB - In atom lithography with optical masks, deposition of an atomic beam on a given substrate is controlled by a standing light-wave field. The lateral intensity distribution of the light field is transferred to the substrate with nanometer scale. We have tailored a complex pattern of this intensity distribution through diffraction of a laser beam from a hologram that is stored in a photorefractive crystal. This method can be extended to superpose 1000 or more laser beams. The method is furthermore applicable during growth processes and thus allows full 3D structuring of suitable materials with periodic and non-periodic patterns at nanometer scales. PMID- 11863955 TI - Hidden coherence along space-time trajectories in parametric wave mixing. AB - We show theoretically that the waves generated through the generic parametric three- or four-wave-mixing processes exhibit, as a general rule, a hidden coherence characterized by skewed coherence lines along specific space-time trajectories. Our study generalizes the concept of coherence in the sense that these previously unrecognized coherent states cannot be described through the standard definitions of spatial and temporal coherence. PMID- 11863956 TI - Growth of precursors in silicon using pseudopotential calculations. AB - Based on the results of pseudopotential calculations, the progression of a single interstitial into small compact clusters and ultimately into chain-like defects is examined. For clusters that consist of more than eight interstitials, the capture of bond-centered interstitials reveals a change in the growth mechanism leading to enhanced stability of clusters. The five-interstitial model is proposed to be a plausible candidate for optically active W centers, observed in ion irradiated silicon. PMID- 11863957 TI - Dynamic structure of He-Ne mixtures by molecular dynamics simulation: from hydrodynamic to fast and slow sound modes. AB - Molecular dynamics (MD) results for the dynamic structure of a He(0.77)Ne(0.23) gas mixture at two densities (15.8 and 36.1 nm(-3)) show a clear crossover from hydrodynamic modes to distinct excitations for the two species. The higher density dispension curve neatly shows high- and low-frequency branches setting on with a rather localized transition. The lower density results agree very well with existing neutron scattering data and, in particular, display hydrodynamic behavior up to k approximately 2 nm(-1), in contrast with the conclusions of previous simulation studies. A smooth transition to fast sound is shown to take place for 2)e(-->)p-->e(prime)pi(+)n in the resonance region. AB - The double spin asymmetry in the (-->)e(-->)p --> e(prime)pi(+)n reaction has been measured for the first time in the resonance region for four-momentum transfer Q2 = 0.35-1.5 GeV(2). Data were taken at Jefferson Lab with the CLAS detector using a 2.6 GeV polarized electron beam incident on a polarized solid NH3 target. Comparison with predictions of phenomenological models shows strong sensitivity to resonance contributions. Helicity-1/2 transitions are found to be dominant in the second and third resonance regions. The measured asymmetry is consistent with a faster rise with Q(2) of the helicity asymmetry A1 for the F(15)(1680) resonance than expected from the analysis of the unpolarized data. PMID- 11863959 TI - Observation of unusual surface ordering in a uniaxial Sm-A phase formed by a highly biaxial compound. AB - We have studied one banana-shaped compound using null-transmission ellipsometry. By studying free-standing films of various thicknesses we confirm that this compound exhibits the Sm-A phase. This is the first banana-shaped compound with a relatively small bend angle (approximately 140 degrees) in which a uniaxial phase has been observed. At the lower temperature end of the Sm-A phase we observe unusual ordering at the surface of the film. PMID- 11863960 TI - High pressure photoinduced ring opening of benzene. AB - The chemical transformation of crystalline benzene into an amorphous solid (a C:H) was induced at high pressure by employing laser light of suitable wavelengths. The reaction was forced to occur at 16 GPa, well below the pressure value (23 GPa) where the reaction normally occurs. Different laser sources were used to tune the pumping wavelength into the red wing of the first excited singlet state S(1)((1)B(2u)) absorption edge. Here the benzene ring is distorted, presenting a greater flexibility which makes the molecule unstable at high pressure. The selective pumping of the S(1) level, in addition to structural considerations, was of paramount importance to clarify the mechanism of the reaction. PMID- 11863961 TI - Identification of the atomic structure of the fivefold surface of an icosahedral Al-Pd-Mn quasicrystal: helium diffraction and scanning tunneling microscopy studies. AB - High resolution He diffraction and scanning tunneling microscopy images of the fivefold surface of a single-grain i-AlPdMn quasicrystal are obtained showing an almost perfect quasicrystal order. Observed configurations can be identified within the framework of polyhedral models. The terrace terminations are found to be Al-rich planes and successions of step heights agree with distances between dense Al planes in the model. This shows the ability of recent 6D polyhedral models to describe real quasicrystalline atomic configurations. PMID- 11863962 TI - Unusual thickness-dependent heat-capacity anomalies in free-standing hexatic liquid-crystal films. AB - High-resolution heat-capacity and optical-reflectivity measurements have been conducted near the smectic-A to hexatic-B transition in thin free-standing films of the liquid-crystal compound 64COOBC. We find an unexpected dependence on film thickness of the integrated magnitude of the heat-capacity anomalies as the films undergo layer-by-layer transitions. We measure the penetration depths of the ordering from the surface and next-to-surface layers which are pertinent to the highly correlated thermal behavior. PMID- 11863964 TI - Critical Casimir effect near the (3)He-(4)He tricritical point. AB - We present capacitance measurements of the equilibrium thickness of (3)He-(4)He mixture films as a function of temperature and concentration. The films are adsorbed on a Cu substrate situated above bulk liquid mixture. As we scan across the tricritical point, we observe a thickening of the film indicating the presence of a repulsive critical Casimir force. PMID- 11863963 TI - Nucleation in systems with elastic forces. AB - Systems with long-range interactions when quenched into a metastable state near the pseudospinodal exhibit nucleation processes that differ from classical nucleation seen near the coexistence curve. In systems with long-range elastic forces the description of the nucleation process can be quite subtle due to the presence of bulk and surface elastic compatibility constraints. We analyze the nucleation process in a simple 2D model with elastic forces and show that the nucleation process generates critical droplets with a different structure than the stable phase. This has implications for nucleation in many crystal-crystal transitions, specifically martensites and shape memory alloys, and for the structure of the final state. PMID- 11863965 TI - Droplet state and the compressibility anomaly in dilute 2D electron systems. AB - We investigate the space distribution of carrier density and the compressibility of two-dimensional (2D) electron systems by using the local density approximation. The strong correlation is simulated by the local exchange and correlation energies. A slowly varied disorder potential is applied to simulate the disorder effect. We show that the compressibility anomaly observed in 2D systems which accompanies the metal-insulator transition can be attributed to the formation of the droplet state due to a disorder effect at low carrier densities. PMID- 11863966 TI - Beltlike C(60)(-) electron spin density distribution in the organic ferromagnet TDAE-C(60). AB - The observed huge increase in the width of the (3C)NMR spectra of TDAE-C(0 )n the middle of the ferromagnetic phase at 10 K is due to a Jahn-Teller distortion of the C(60 )(-) which becomes visible in view of the resulting changes in the Fermi contact electron-(13)C NMR shifts. The shape of the (13)C spectra allows for a direct determination of the belt-like redistribution of the unpaired electron spin density on the C(60)(-) ions, which is responsible for the relatively high ferromagnetic transition temperature in this purely organic system. PMID- 11863967 TI - First-principles theory of the EPR g tensor in solids: defects in quartz. AB - A theory for the reliable prediction of the EPR g tensor for paramagnetic defects in solids is presented. It is based on density functional theory and on the gauge including projector augmented wave approach to the calculation of all-electron magnetic response. The method is validated by comparison with existing quantum chemical and experimental data for a selection of diatomic radicals. We then perform the first prediction of EPR g tensors in the solid state and find the results to be in excellent agreement with experiment for the E(1)(prime) and substitutional phosphorus defect centers in quartz. PMID- 11863968 TI - Coherent Hall effect in a semiconductor superlattice. AB - The coherent Hall effect denotes the transient Hall response of impulsively excited coherent charge-carrier wave packets in a solid. We report the first experimental study of this phenomenon (i) using a semiconductor superlattice in crossed electric and magnetic fields as a model for three-dimensional materials and (ii) employing a contactless optoelectronic technique to probe the transient currents. Two field regimes with distinctly different oscillatory wave packet dynamics are found, separated from each other by a transition region where all oscillations are suppressed. PMID- 11863969 TI - Superconducting gap in MgB(2): electronic Raman scattering measurements of single crystals. AB - Polarization-resolved Raman scattering measurements were performed on MgB(2) single crystals to determine the magnitude, symmetry, and temperature dependence of the superconducting gap. A single sharp peak due to Cooper pair breaking appears in the electronic continuum below T(c), reaching a maximum Raman shift of 105 +/- 1 cm(-1) [2 Delta(0)/k(B)T(c) = 3.96 +/- 0.09] and showing up to 5 cm(-1) anisotropy between polarized and depolarized spectra. The temperature dependence of 2 Delta follows that predicted from BCS theory, while the anisotropy decreases with decreasing temperature. It is concluded that the Raman results are consistent with a slightly anisotropic s-wave gap in a conventional BCS superconductor. PMID- 11863970 TI - Driven dynamics of the vortex-phase mixture near the peak effect: the "vortex capacitor". AB - A phenomenological model of vortex dynamics in the vicinity of the peak effect, which considers a coexistence of two vortex phases with different critical current densities, is proposed. It provides a quantitative description of the widely reported anomalous dynamic effects in terms of "contamination" and "annealing" of the vortex matter. Scanning ac Hall microscopy in the superconductor NbSe(2) is used to obtain the first real-space visualization of pinning in the driven vortex lattice. The results are consistent with the proposed scenario. PMID- 11863971 TI - Aging in the random energy model. AB - The random energy model (REM) has become a key reference model for glassy systems. In particular, it is expected to provide a prime example of a system whose dynamics shows aging, a universal phenomenon characterizing the dynamics of complex systems. The analysis of its activated dynamics is based on so-called trap models, introduced by Bouchaud, that are also used to mimic the dynamics of more complex disordered systems. In this Letter we report the first results that justify rigorously the trap model predictions in the REM. PMID- 11863972 TI - Magnetic domain confinement by anisotropy modulation. AB - The spin configuration in a magnet is in general a "natural" consequence of both the intrinsic properties of the material and the sample dimensions. We demonstrate that this limitation can be overcome in a homogeneous ferromagnetic film by engineering an anisotropy contrast. Substrates with laterally modulated single-crystal and polycrystalline surface regions were used to induce selective epitaxial growth of a ferromagnetic Ni film. The resulting spatially varying magnetic anisotropy leads to regular perpendicular and in-plane magnetic domains, separated by a new type of magnetic wall---the "anisotropy constrained" magnetic wall. PMID- 11863973 TI - Interplay of Rabi oscillations and quantum interference in semiconductor quantum dots. AB - We investigated the manifestation of Rabi oscillation in the coherent dynamics of excitons in self-assembled semiconductor quantum dots. The Rabi oscillation phenomenon was directly observed as a function of the input pulse area. Furthermore, by performing wave packet interferometry in the nonlinear excitation regime, we discover a new type of quantum interference phenomenon, resulting from the interplay between Rabi oscillation and quantum interference. PMID- 11863974 TI - Tilt texture domains on a membrane and chirality induced budding. AB - We study the equilibrium conformations of a lipid domain on a planar fluid membrane where the domain is decorated by a vector field representing the tilt of the stiff fatty acid chains of the lipid molecules, while the surrounding membrane is fluid and structureless. The inclusion of chirality in the bulk of the domain induces a novel budding of the membrane, which preempts the budding induced by a decrease in interfacial tension. PMID- 11863975 TI - Neural network model for apparent deterministic chaos in spontaneously bursting hippocampal slices. AB - A neural network model that exhibits stochastic population bursting is studied by simulation. First return maps of interburst intervals exhibit recurrent unstable periodic-orbit(UPO)-like trajectories similar to those found in experiments on hippocampal slices. Applications of various control methods and surrogate analysis for UPO detection also yield results similar to those of experiments. Our results question the interpretation of the experimental data as evidence for deterministic chaos and suggest caution in the use of UPO-based methods for detecting determinism in time-series data. PMID- 11863976 TI - Direct optical quantification of backflow in a 90 degrees twisted nematic cell. AB - Optical guided mode observations of the transient director profile (optical tensor distribution) during the relaxation of a 90 degrees twisted nematic cell directly reveals backflow. In the first 6 ms of the relaxation process, after a voltage across the cell is removed, the midplane tilt of the director increases, reaching a maximum value of 101 degrees at 1.4 ms. This increase in midplane tilt is attributed to coupling between fluid flow (backflow) and director reorientation. A 270 degrees twisted state of the opposite handedness to the 90 degrees twisted state found at equilibrium is shown to exist during the backflow period. Good fits of theoretical models with experimentally determined time dependent director profiles yield the viscosity coefficients. PMID- 11863977 TI - Propagation of hydrodynamic interactions in colloidal suspensions. AB - We describe direct measurements of the dynamics of two colloidal spheres before hydrodynamic interactions have had time to fully develop. We find that the dynamics of the two spheres are coupled at times significantly shorter than tau(nu), the time required for vorticity to diffuse between the two spheres. From the distance dependence of the measured coupling, we infer that hydrodynamic interactions develop in a sonic time scale. PMID- 11863978 TI - Packet waves in a reaction-diffusion system. AB - The finite-wavelength instability gives rise to a new type of wave in reaction diffusion systems: packet waves, which propagate only within a wave packet, are found in experiments on the Belousov-Zhabotinsky reaction dispersed in water-in oil AOT microemulsion (BZ-AOT) as well as in model simulations. Inwardly moving packet waves with negative curvature occur in experiments and in a model of the BZ-AOT system when the dispersion d omega(k)/dk is negative at the characteristic wave number k(0). This result sheds light on the origin of anti-spirals. PMID- 11863979 TI - Enhanced drainage and coarsening in aqueous foams. AB - Experiments are presented elucidating how the evolution of foam microstructure by gas diffusion from high to low pressure bubbles can significantly speed up the rate of gravitational drainage, and vice versa. This includes detailed data on the liquid-fraction dependence of the coarsening rate, and on the liquid-fraction and the bubble-size profiles across a sample. These results can be described by a "coarsening equation" for the increase of bubble growth rate for drier foams. Spatial variation of the average bubble size and liquid fraction can also affect the growth and drainage rates. PMID- 11863980 TI - Comment on "A convergent series for the QED effective action". PMID- 11863981 TI - Comment on "Novel phase behavior in a two-dimensional network-forming lattice fluid". PMID- 11863982 TI - Comment on "Elasticity model of a supercoiled DNA molecule". PMID- 11863984 TI - Periodically dressed Bose-Einstein condensate: a superfluid with an anisotropic and variable critical velocity. AB - We consider a two-component atomic gas illumined by two intersecting laser beams which induce Raman coupling between the components. This spatially periodic coupling modifies the dispersion relation of the gas. Properties of a Bose Einstein condensate of such a gas are strongly affected by this modification. Using the quasiparticle excitation spectrum derived from a Bogoliubov transformation, the Landau critical velocity is found to be anisotropic and can be widely tuned by varying properties of the dressing laser beams. PMID- 11863985 TI - Signatures of resonance superfluidity in a quantum Fermi gas. AB - We predict a direct and observable signature of the superfluid phase in a quantum Fermi gas, in a temperature regime already accessible in current experiments. We apply the theory of resonance superfluidity to a gas confined in a harmonic potential and demonstrate that a significant increase in density will be observed in the vicinity of the trap center. PMID- 11863986 TI - Rate limit for photoassociation of a Bose-Einstein condensate. AB - We simulate numerically the photodissociation of molecules into noncondensate atom pairs that accompanies photoassociation of an atomic Bose-Einstein condensate into a molecular condensate. Such rogue photodissociation sets a limit on the achievable rate of photoassociation. Given the atom density rho and mass m, the limit is approximately 6(planck)rho(2/3)/m. PMID- 11863987 TI - Creation of a monopole in a spinor condensate. AB - We propose a method to create a monopole structure in a multicomponent condensate by applying the basic methods used to create vortices and solitons experimentally in single-component condensates. We also show that by using a two-component structure for a monopole, we can avoid many problems related to the previously suggested three-component monopole. We discuss the observation and dynamics of such a monopole structure, and note that the dynamics of the two-component monopole differs from the dynamics of the three-component monopole. PMID- 11863988 TI - Clock-comparison tests of Lorentz and CPT symmetry in space. AB - Clock-comparison experiments conducted in space can provide access to many unmeasured coefficients for Lorentz and CPT violation. The orbital configuration of a satellite platform and the relatively large velocities attainable in a deep space mission would permit a broad range of tests with Planck-scale sensitivity. PMID- 11863989 TI - Calculating the gravitational self-force in Schwarzschild spacetime. AB - We present a practical method for calculating the local gravitational self-force (often called "radiation-reaction force") for a pointlike particle orbiting a Schwarzschild black hole. This is an implementation of the method of mode-sum regularization, in which one first calculates the (finite) contribution to the force due to each individual multipole mode of the perturbation, and then applies a certain regularization procedure to the mode sum. Here we give the values of all the "regularization parameters" required for implementing this regularization procedure, for any geodesic orbit in Schwarzschild spacetime. PMID- 11863990 TI - Tachyon condensation and black hole entropy. AB - String propagation on a cone with deficit angle 2pi(1-1 / N) is considered for the purpose of computing the entropy of a large mass black hole. The entropy computed using the recent results on condensation of twisted-sector tachyons in this theory is found to be in precise agreement with the Bekenstein-Hawking entropy. PMID- 11863991 TI - Probing the dark energy with quasar clustering. AB - We show through Monte Carlo simulations that the Alcock-Paczynski test, as applied to quasar clustering, is a powerful tool to probe the cosmological density and equation of state parameters Omega(m0), Omega(x0), and w. By taking into account the effect of peculiar velocities upon the correlation function we obtain for the Two-Degree Field QSO Redshift Survey the predicted confidence contours for the cosmological constant (w = -1) and spatially flat (Omega(m0)+Omega(x0) = 1) cases. For w = -1, the test is especially sensitive to the difference Omega(m0)-Omega(Lambda0), thus being ideal to combine with cosmic microwave background results. For the flat case, it is competitive with future supernova and galaxy number count tests, besides being complementary to them. PMID- 11863992 TI - Changing alpha with time: implications for fifth-force-type experiments and quintessence. AB - If the recent observations suggesting a time variation of the fine structure constant are correct, they imply the existence of an ultralight scalar particle. This particle inevitably couples to nucleons through the alpha dependence of their masses and thus mediates an isotope-dependent long-range force. The strength of the coupling is within a couple of orders of magnitude of the existing experimental bounds for such forces. The new force can be potentially measured in precision experimental tests of the equivalence principle. Because of a coincidence of the required time scales, the scalar field can at the same time play the role of a quintessence field. PMID- 11863993 TI - Thermally generated gauge singlet scalars as self-interacting dark matter. AB - We show that a gauge singlet scalar S, with a coupling to the Higgs doublet of the form lambda(S)S+SH+H and with the S mass entirely generated by the Higgs expectation value, has a thermally generated relic density Omega(S)approximately equal to 0.3 if m(S)approximately equal to (2.9-10.5) (Omega(S)/0.3)(1/5)(h/0.7)(2/5) MeV. Remarkably, this is very similar to the range [m(S) = (6.6-15.4)eta(2/3) MeV] required in order for the self-interaction (eta/4) (S+S)(2) to account for self-interacting dark matter when eta is not much smaller than 1. The corresponding coupling is lambda(S)approximate(2.7 x 10(-10) 3.6 x 10(-9)) (Omega(S)/0.3)(2/5)(h/0.7)(4/5), implying that such scalars are very weakly coupled to the standard model sector. PMID- 11863994 TI - Recoil correction to the bound-electron g factor in H-like atoms to all orders in alphaZ. AB - The nuclear-recoil correction to the bound-electron g factor in H-like atoms is calculated to first order in m/M and to all orders in alphaZ. The calculation is performed in the range Z = 1--100. A large contribution of terms of order (alphaZ)(5) and higher is found. Even for hydrogen, the higher-order correction exceeds the (alphaZ)(4) term, while for uranium it is above the leading (alphaZ)(2) correction. As a result, one of the main sources of the theoretical uncertainty for the bound-electron g factor is eliminated. PMID- 11863995 TI - Precise determination of electroweak parameters in neutrino-nucleon scattering. AB - The NuTeV Collaboration has extracted the electroweak parameter sin(2)theta(W) from the measurement of the ratios of neutral current to charged current nu and ( )nu cross sections. Our value, sin(2)theta((on-shell))(W) = 0.2277 +/- 0.0013(stat) +/- 0.0009(syst), is 3 standard deviations above the standard model prediction. We also present a model independent analysis of the same data in terms of neutral-current quark couplings. PMID- 11863998 TI - Nuclear masses: evidence of order-chaos coexistence. AB - Shell corrections are important in the determination of nuclear ground-state masses and shapes. Although general arguments favor a regular single-particle dynamics, symmetry breaking and the presence of chaotic layers cannot be excluded. The latter provide a natural framework that explains the observed differences between experimental and computed masses. PMID- 11863999 TI - K-shell photodetachment of He- : experiment and theory. AB - High-resolution K-shell photodetachment measurements of He- giving rise to He+ ions have been performed using a merged synchrotron vacuum ultraviolet photon-ion beam technique. The measurements on this fundamental negative ion display dramatic structure differing substantially, qualitatively and quantitatively, from the corresponding process in neutral atoms and positive ions, owing to the dominance of correlation in both initial and final states of He-. In addition, this experimental investigation provides an unambiguous test of two independent theoretical calculations that report serious discrepancies and shows excellent agreement with one of them. PMID- 11863996 TI - Measurement of G(E(p))/G(M(p)) in e(-->)p---> e(-->)p to Q(2) = 5.6 GeV(2). AB - The ratio of the electric and magnetic form factors of the proton G(E(p))/G(M(p)), which is an image of its charge and magnetization distributions, was measured at the Thomas Jefferson National Accelerator Facility (JLab) using the recoil polarization technique. The ratio of the form factors is directly proportional to the ratio of the transverse to longitudinal components of the polarization of the recoil proton in the elastic e(-->)p---> e(-->)p reaction. The new data presented span the range 3.5< Q(2)< 5.6 GeV(2) and are well described by a linear Q(2) fit. Also, the ratio sqrt[Q(2)] F(2(p))/F(1(p)) reaches a constant value above Q(2) = 2 GeV(2). PMID- 11864000 TI - Strong electron correlation in photoionization of spin-orbit doublets. AB - A new and explicitly many-body aspect of the "leveraging" of the spin-orbit interaction is demonstrated, spin-orbit activated interchannel coupling, which can significantly alter the photoionization cross section of a spin-orbit doublet. As an example, it is demonstrated via a modified version of the spin polarized random phase approximation with exchange, that a recently observed unexplained structure in the Xe 3d(5/2) photoionization cross section [A. Kivimaki et al., Phys. Rev. A 63, 012716 (2000)] is entirely due to this effect. Similar features are predicted for Cs 3d(5/2) and Ba 3d(5/2). PMID- 11863997 TI - (68)(28)Ni(40): Magicity versus superfluidity. AB - The neutron-rich (66,68)Ni have been produced at GANIL via interactions of a 65.9A MeV 70Zn beam with a 58Ni target. Their reduced transition probability B(E2;0(+)(1)-->2+) has been measured for the first time by Coulomb excitation in a (208)Pb target at intermediate energy. The B(E2) value for (68)Ni(40) is unexpectedly small. An analysis in terms of large scale shell model calculations stresses the importance of proton core excitations to reproduce the B(E2) values and indicates the erosion of the N = 40 harmonic-oscillator subshell by neutron pair scattering. PMID- 11864001 TI - Feedback control of atomic motion in an optical lattice. AB - We demonstrate a real-time feedback scheme to manipulate wave-packet oscillations of atoms in an optical lattice. The average position of the atoms in the lattice wells is measured continuously and nondestructively. A feedback loop processes the position signal and translates the lattice potential. Depending on the feedback loop characteristics, we find amplification, damping, or an entire alteration of the wave-packet oscillations. Our results are well supported by simulations. PMID- 11864002 TI - Magnetic slowing down of spin relaxation due to binary collisions of alkali-metal atoms with buffer-gas atoms. AB - We report the first studies of magnetic decoupling of the spin relaxation of alkali-metal atoms due to binary collisions with buffer gases. When binary collisions are the dominant relaxation mechanism, the relaxation and its magnetic decoupling are well described by the S-damping rate Gamma(SD) due to the spin rotation interaction gammaN.S, the spin exchange rate Gamma(EX) for collisions between alkali atoms, and a new "Carver rate" Gamma(C), due to the pressure-shift interaction deltaAI.S, which can substantially broaden the magnetic decoupling curve while having no influence on the zero-field rates. PMID- 11864003 TI - Interisotope determination of ultracold rubidium interactions from three high precision experiments. AB - Combining the measured binding energies of four of the most weakly bound rovibrational levels of the 87Rb2 molecule with results of two other recent high precision experiments, we obtain exceptionally strong constraints on the atomic interaction parameters in a highly model independent analysis. The comparison of (85)Rb and (87)Rb data, where the two isotopes are related by a mass scaling procedure, plays a crucial role. We predict scattering lengths, clock shifts, and Feshbach resonances with an unprecedented level of accuracy. Two of the Feshbach resonances occur at easily accessible magnetic fields in mixed-spin channels. One is related to a d-wave shape resonance. PMID- 11864004 TI - Polarization squeezing of continuous variable stokes parameters. AB - We report the first direct experimental characterization of continuous variable quantum Stokes parameters. We generate a continuous wave light beam with more than 3 dB of simultaneous squeezing in three of the four Stokes parameters. The polarization squeezed beam is produced by mixing two quadrature squeezed beams on a polarizing beam splitter. Depending on the squeezed quadrature of these two beams the quantum uncertainty volume on the Poincare sphere becomes a "cigarlike" or "pancakelike" ellipsoid. PMID- 11864005 TI - Anomalous refraction and diffraction in discrete optical systems. AB - We experimentally prove that light propagation in a discrete system, i.e., an array of coupled waveguides, exhibits striking anomalies. We show that refraction is restricted to a cone, irrespective of the initial tilt of the beam. Diffraction can be controlled in size and sign by the input conditions. Diffractive beam spreading can even be arrested and diverging light can be focused. The results can be thoroughly theoretically explained. PMID- 11864006 TI - Decay distribution of spontaneous emission from an assembly of atoms in photonic crystals with pseudogaps. AB - Spontaneous decay behaviors from an assembly of atoms (or molecules) in 3D photonic crystals (PC's) with pseudogaps are investigated. Theoretically, a lifetime distribution function for these atoms or molecules is defined to reveal decay kinetics. Our calculations show that quite wide or narrow lifetime distributions can occur for different spread configurations of the atoms (or molecules). The pure PC effect may lead to coexistence of both accelerated and inhibited decay processes. These results provide theoretical clarification for substantial discrepancies in the recent reported experiments. PMID- 11864007 TI - Autonomous swinging of a lipid tubule under stationary irradiation by a Nd3+:YAG laser. AB - A micrometer-sized lipid tubule exhibits oscillatory swinging motion under stationary irradiation by a Nd(3+):YAG laser ( lambda = 1064 nm). By choosing an appropriate optical path through an objective lens, the laser can be split into dual beams focused on the same position. Using this splitting, a lipid tubule can be shown to exhibit bistability with regard to the orientation of trapping. Driven by a temperature gradient induced by local photon heating, the trapped lipid tubule shows oscillatory motion between two possible orientations. This oscillatory phenomenon of the lipid tubule is thought to represent the breaking of time-translational symmetry under thermodynamically open conditions. PMID- 11864008 TI - Random laser in one dimension. AB - We present an analytical approach to random lasing in a one-dimensional medium, consistent with transfer matrix numerical simulations. It is demonstrated that the lasing threshold is defined by transmission through the passive medium and thus depends exponentially on the size of the system. Lasing in the most efficient regime of strong three-dimensional localization of light is discussed. We argue that the lasing threshold should have anomalously strong fluctuations from probe to probe, in agreement with recent measurements. PMID- 11864009 TI - Numerical characterization of high harmonic attosecond pulses. AB - A numerical simulation of attosecond harmonic pulse generation in a three dimensional field-ionizing gas is presented. Calculated harmonic efficiencies quantitatively reproduce experimental findings. This allows a quantitative characterization of attosecond pulse generation revealing information currently not accessible by experiment. The rapid phase variation and spatiotemporal distortions of harmonics are smaller than anticipated, allowing focusing of 30 nm, 750-as pulses to intensities in excess of 10(13) W/cm(2). Feasibility of such pulses brings novel applications such as extreme ultraviolet nonlinear optics and attosecond pump probe spectroscopy within reach. PMID- 11864010 TI - Quantum dissipative chaos in the statistics of excitation numbers. AB - A quantum manifestation of chaotic classical dynamics is found in the framework of oscillatory number statistics for the model of a nonlinear dissipative oscillator. It is shown that the probability distributions and variances of oscillatory number states are strongly transformed in the order-to-chaos transition. A nonclassical, sub-Poissonian statistics of oscillatory excitation numbers is established for chaotic dissipative dynamics in the framework of the Fano factor and Wigner functions. It is proposed to use these results in experimental studies of the quantum dissipative chaos. PMID- 11864011 TI - Fresnel filtering in lasing emission from scarred modes of wave-chaotic optical resonators. AB - We study lasing emission from asymmetric resonant cavity GaN microlasers. By comparing far-field intensity patterns with images of the microlaser we find that the lasing modes are concentrated on three-bounce unstable periodic ray orbits; i.e., the modes are scarred. The high-intensity emission directions of these scarred modes are completely different from those predicted by applying Snell's law to the ray orbit. This effect is due to the process of "Fresnel filtering" which occurs when a beam of finite angular spread is incident at the critical angle for total internal reflection. PMID- 11864012 TI - Finite heat conduction in a 2D disorder lattice. AB - Recently, the thermal conductivity in disordered chains has attracted a lot of attention, including the discussion of the disorder anharmonic and harmonic chains. A natural problem concerns the higher-dimensional disordered lattice system. This paper gives a 2D harmonic lattice model with missing bond defects. When the defect density is large enough, a temperature gradient builds up and a finite heat conduction is found in this model. PMID- 11864014 TI - Relative dispersion in fully developed turbulence: the Richardson's law and intermittency corrections. AB - Relative dispersion in fully developed turbulence is investigated by means of direct numerical simulations. Lagrangian statistics is found to be compatible with Richardson description although small systematic deviations are found. The value of the Richardson constant is estimated as C2 approximately equal to 0.55, in a close agreement with recent experimental findings [S. Ott and J. Mann, J. Fluid Mech. 422, 207 (2000)]. By means of exit-time statistics it is shown that the deviations from Richardson's law are a consequence of Eulerian intermittency. The measured Lagrangian scaling exponents require a set of Eulerian structure function exponents zeta(p) which are remarkably close to standard ones known for fully developed turbulence. PMID- 11864013 TI - Controlling the energy flow in nonlinear lattices: a model for a thermal rectifier. AB - We address the problem of heat conduction in 1D nonlinear chains; we show that, acting on the parameter which controls the strength of the on-site potential inside a segment of the chain, we induce a transition from conducting to insulating behavior in the whole system. Quite remarkably, the same transition can be observed by increasing the temperatures of the thermal baths at both ends of the chain by the same amount. The control of heat conduction by nonlinearity opens the possibility to propose new devices such as a thermal rectifier. PMID- 11864015 TI - On-line ion cooling and bunching for collinear laser spectroscopy. AB - A new method has been developed for increasing the sensitivity of collinear laser spectroscopy. The method utilizes an ion-trapping technique in which a continuous low-energy ion beam is cooled and accumulated in a linear Paul trap and subsequently released as a short (10-20 micros) bunch. In collinear laser measurements the signal-to-noise ratio has been improved by a factor of 2 x 10(4), allowing spectroscopic measurements to be made with ion-beam fluxes of approximately 50 ions s(-1). The bunching method has been demonstrated in an on line isotope shift and hyperfine structure measurement on radioactive (175)Hf. PMID- 11864016 TI - Variational principles and self-organization in two-fluid plasmas. AB - Self-organization of an ordered structure occurs in a plasma under rather restrictive conditions. A new framework for a variational principle invokes a coercive form that results in a criterion for self-organizing relaxation of a two fluid plasma. The constraints (constants of motion of the ideal model) are adjusted, through a weakly dissipative process, so that the relaxed state, under well-defined conditions, is a stable equilibrium independent of the direct effects of dissipation. PMID- 11864017 TI - Electron cyclotron resonance in a weakly magnetized radio-frequency inductive discharge. AB - The evolution of the electron energy distribution function (EEDF) over a weak magnetic field range is investigated in magnetized radio-frequency (rf) inductive discharges under a collisionless regime where an anomalous skin effect and electron cyclotron resonance (ECR) can occur. A significant change in the low energy range of the EEDF is found in the ECR condition during the evolution. The observed result reveals the low-energy electrons are efficiently heated by the rf ECR in the presence of the anomalous skin effect. The calculated result of the electron distribution based on kinetic theory is in good agreement with the experiment. PMID- 11864018 TI - Electromagnetically induced transparency in ensembles of classical oscillators. AB - We develop a classical model of the parametric effect of electromagnetically induced transparency (EIT) within the line of resonance absorption of an electromagnetic wave in the medium--an effect initially discovered for a quantum three-level system. On the basis of this model, the EIT effect for electromagnetic waves at frequencies of the electron-cyclotron resonance in a cold plasma is considered. Similar to the analogous quantum scheme, the EIT window in the classical model is characterized by group deceleration of the reference electron-cyclotron wave. PMID- 11864019 TI - Dissipative longitudinal solitons in a two-dimensional strongly coupled complex (dusty) plasma. AB - Solitary waves are experimentally studied in a monolayer hexagonal dust lattice which is formed from monodisperse plastic microspheres and levitated in the sheath of an rf discharge. It is found that the product of the soliton amplitude and the square of the soliton width is constant as the soliton propagates. The analytical theory describing the experiment is based on the equations of motion written for a linear chain. It takes into account damping, dispersion, and nonlinearity. The numerical simulation of a linear chain produces double solitons like those observed in the experiment. PMID- 11864020 TI - Electron acceleration by a tightly focused laser beam. AB - State-of-the-art petawatt laser beams may be focused down to few-micron spot sizes and can produce violent electron acceleration as a result of the extremely intense and asymmetric fields. Classical fifth-order calculations in the diffraction angle show that electrons, injected sideways into the tightly focused laser beam, get captured and gain energy in the GeV regime. We point out the most favorable points of injection away from the focus, along with an efficient means of extracting the energetic electron with a static magnetic field. PMID- 11864021 TI - Instability-triggered phase transition to a dusty-plasma condensate. AB - Highly charged dust grains in plasma discharges reside at the sheath edge, where the ions stream toward the electrode at speed approximately c(s). Above a critical pressure P(crit), the grains lose their kinetic energy and reach a strongly coupled crystalline state, but for P P(crit) by the combined effect of ion-molecule and grain-molecule collisions. PMID- 11864022 TI - Origins of coexistence of conductivity and transparency in SnO(2). AB - SnO2 is a prototype "transparent conductor," exhibiting the contradictory properties of high metallic conductivity due to massive structural nonstoichiometry with nearly complete, insulator-like transparency in the visible range. We found, via first-principles calculations, that the tin interstitial and oxygen vacancy have surprisingly low formation energies and strong mutual attraction, explaining the natural nonstoichiometry of this system. The stability of these intrinsic defects is traced back to the multivalence of tin. These defects donate electrons to the conduction band without increasing optical interband absorption, explaining coexistence of conductivity with transparency. PMID- 11864023 TI - Spiral cracks in drying precipitates. AB - We investigate the formation of spiral crack patterns during the desiccation of thin layers of precipitates in contact with a substrate. This symmetry-breaking fracturing mode is found to arise naturally not from torsion forces but from a propagating stress front induced by the foldup of the fragments. We model their formation mechanism using a coarse-grain model for fragmentation and successfully reproduce the spiral cracks. Fittings of experimental and simulation data show that the spirals are logarithmic. Theoretical aspects of the logarithmic spirals are discussed. In particular we show that this occurs generally when the crack speed is proportional to the propagating speed of stress front. PMID- 11864024 TI - Beyond Flory-Huggins theory: new classes of blend miscibility associated with monomer structural asymmetry. AB - Flory-Huggins (FH) theory is restricted to polymer mixtures whose monomers are structurally identical, a situation limited to isotopic blends and computer simulations. We investigate the influence of monomer structure on blend miscibility and scattering properties using the lattice cluster theory generalization of the FH model. Monomer structural asymmetry is shown to profoundly affect blend miscibility (T(c),phi(c)), chain swelling (T(theta)), and the scale (xi) and intensity [S(0)] of composition fluctuations. Four distinct blend miscibility classes are identified and experimental evidence for these classes is discussed. PMID- 11864025 TI - Phase equilibria of charge-, size-, and shape-asymmetric model electrolytes. AB - The low-temperature phase behavior of two 2:1 hard-core electrolyte models has been investigated by Monte Carlo simulations. In the first model, both bivalent cations and monovalent anions are spherical, and the charges are located at the ion's centers; in the second model, bivalent cations are modeled as rigid dimers composed of two tangent hard spheres, each carrying a positive charge at the center. It is found that the critical temperature and the critical density are strongly affected by the size asymmetry and the shape of the ions. The results presented in this work provide insights into the behavior of charged colloidal suspensions and polyelectrolytes, where large, symmetric or asymmetric ionic species carrying like charges can attract each other and give rise to thermodynamically unstable conditions. PMID- 11864026 TI - From strong to fragile glass formers: secondary relaxation in polyalcohols. AB - We have studied details of the molecular origin of slow secondary relaxation near T(g) in a series of neat polyalcohols by means of dielectric spectroscopy and (2)H NMR. From glycerol to threitol, xylitol, and sorbitol the appearance of the secondary relaxation changes gradually from a wing-type scenario to a pronounced beta peak. It is found that in sorbitol the dynamics of the whole molecule contributes equally to the beta process, while in glycerol the hydrogen bond forming OH groups remain rather rigid compared to the hydrogens bonded to the carbon skeleton. PMID- 11864027 TI - Singularity in the thermal boundary resistance between superfluid (4)He and a solid surface. AB - We report new measurements in four cells of the thermal boundary resistance R between copper and (4)He below but near the superfluid-transition temperature T(lambda). For 10(-7)< or =t identical to 1-T/T(lambda))< or =10(-4) fits of R = R(0)t(-x(b))+R(B) to the data yielded x(b) approximately equal to 0.18, whereas a fit to theoretical values based on the renormalization-group theory yielded x(b) = 0.23. Alternatively, a good fit of the theory to the data could be obtained if the amplitude of the prediction was reduced by a factor close to 2. The results raise the question whether the boundary conditions used in the theory should be modified. PMID- 11864028 TI - Magnetic field enhancement of heat transport in the 2D Heisenberg antiferromagnet K2V3O8. AB - The thermal conductivity and heat capacity of single crystals of the spin 1/2 quasi-2D Heisenberg antiferromagnet K(2)V(3)O(8) have been measured from 1.9 to 300 K in magnetic fields from 0 to 8 T. The zero field thermal conductivity data are consistent with resonant scattering of phonons by magnons near the zone boundary. Application of a magnetic field greater than 1 T, however, produces a new magnetic ground state with substantial heat transport by long wavelength magnons. PMID- 11864029 TI - Ionic self-diffusion in concentrated aqueous electrolyte solutions. AB - A self-consistent microscopic theory is developed to understand the anomalously weak concentration dependence of ionic self-diffusion coefficient D(ion) in electrolyte solutions. The self-consistent equations are solved by using the mean spherical approximation expressions of the static pair correlation functions for unequal sizes. The results are in excellent agreement both with the known experimental results for many binary electrolytes and also with the new Brownian dynamics simulation results. The calculated velocity time correlation functions also show quantitative agreement with simulations. The theory also explains the reason for observing different D(ion) in recent NMR and neutron scattering experiments. PMID- 11864030 TI - Elastic relaxations in ultrathin epitaxial alloy films. AB - Elastic interactions responsible for the stability of nanometer-scale patterns in ultrathin, bulk-immiscible-alloy films are analyzed within the context of a hybrid atomistic-continuum model. Two apparently different descriptions of alloy film behavior, a continuum elasticity theory describing a deformable substrate and a rigid substrate atomistic scheme, emerge naturally as limiting cases on long and short length scales, respectively. Quantitative first-principles calculations explain the origin of recently observed nanoscale patterns in Co Ag/Ru(0001), and reveal a surprising failure of the continuum model. PMID- 11864031 TI - Transition from stick-slip to smooth sliding: an earthquakelike model. AB - We present a detailed study of an earthquakelike model that exhibits a "transition" from stick-slip motion to smooth sliding at a velocity of the order of those observed in experiments. This contrasts with the many previous microscopic models in which the transition velocity is many orders of magnitude too large. The results show that experimentally observed smooth sliding at the macroscopic scale must correspond to microscopic-scale stick-slip motion. PMID- 11864032 TI - Thickness induced structural transition in suspended fcc metal nanofilms. AB - Recent experiments show that at a critical thickness, surface forces can dominate the bulk coercing suspended Au nanofilms to globally reconstruct from the (001) to the (111) orientation. Here we perform molecular dynamics simulations demonstrating that such transformations are generic to other fcc metals. This contrasts with surface reconstruction on the bulk occurring only in 5d metals. We show that this phenomenon occurs once a small energy barrier is overcome and discuss the relation of such a barrier and the critical thickness to film surface area and boundary conditions. PMID- 11864033 TI - Structure of quantum wires in Au/Si(557). AB - The structure of the Au/Si(557) surface is determined from three-dimensional x ray diffraction measurements, which directly mandate a single Au atom per unit cell. We use a "heavy atom" method in which the Au atom images the rest of the structure. Au is found to substitute for a row of first-layer Si atoms in the middle of the terrace, which then reconstructs by step rebonding and adatoms. The structure is consistent with the 1D metallic behavior seen by photoemission. PMID- 11864034 TI - Self-organized nanoscale pattern formation on vicinal Si(111) surfaces via a two stage faceting transition. AB - We demonstrate a self-organized pattern formation on vicinal Si(111) surfaces that are miscut toward the [2;11] direction. All the patterns, consisting of a periodic array of alternating (7 x 7) reconstructed terraces and step-bunched facets, have the same periodicity and facet structure, independent of the miscut angle, while the width of the facets increases linearly with miscut angle. We attribute such unique pattern formation to a surface faceting transition that involves two transition stages: the first stage forms a stress-domain structure defining the universal periodicity; the second stage forms the low-energy facets controlling the facet width. PMID- 11864035 TI - Evidence for insulating behavior in the electric conduction of (NH(3))K(3)C(60) systems. AB - Microwave study using the cavity perturbation technique revealed that the conductivity of the antiferromagnet (NH(3))K(3-x)Rb(x)C(60) at 200 K is already 3 4 orders of magnitude smaller than those of superconductors, K(3)C(60) and (NH(3))(x)NaRb(2)C(60), and that the antiferromagnetic compounds are insulators below 250 K without metal-insulator transitions. The striking difference in the magnitude of the conductivity between these materials strongly suggests that the Mott-Hubbard transition in the ammoniated alkali fullerides is driven by a reduction of lattice symmetry from face-centered-cubic to face-centered orthorhombic, rather than by the magnetic ordering. PMID- 11864036 TI - Spectroscopic signatures of spin-charge separation in the quasi-one-dimensional organic conductor TTF-TCNQ. AB - The electronic structure of the quasi-one-dimensional organic conductor TTF-TCNQ is studied by angle-resolved photoelectron spectroscopy (ARPES). The experimental spectra reveal significant discrepancies to band theory. We demonstrate that the measured dispersions can be consistently mapped onto the one-dimensional Hubbard model at finite doping. This interpretation is further supported by a remarkable transfer of spectral weight as a function of temperature. The ARPES data thus show spectroscopic signatures of spin-charge separation on an energy scale of the conduction bandwidth. PMID- 11864037 TI - Spectral function of a quarter-filled one-dimensional charge density wave insulator. AB - We consider a one-dimensional charge density wave insulator formed by umklapp processes in a quarter-filled band. The spectrum of the model consists of gapless, uncharged excitations carrying spin +/- 1/2 (spinons) and gapped, spinless excitations carrying charge -/+ signe/2 (solitons and antisolitons). We calculate the low-energy behavior of the single-electron Green's function at zero temperature. The spectral function exhibits a featureless scattering continuum of two solitons and many spinons. The theory predicts that the gap observed by angle resolved photoemission is twice the activation gap in the dc conductivity. We comment on possible applications to PrBa(2)Cu(3)O(7) and to the Bechgaard salts. PMID- 11864038 TI - Nonuniversal behavior of scattering between fractional quantum Hall edges. AB - In a fractional quantum Hall system with a narrow constriction, tunneling of quasiparticles between states at different edges can lead to resistance and to shot noise. The ratio of the shot noise to the backscattered current, in the weak scattering regime, measures the fractional charge of the quasiparticle, which has been confirmed in several experiments. However, the predicted nonlinearity of the resistance was apparently not observed in some of these cases. As a possible explanation, we consider a model where coupling between the current carrying edge mode and additional phononlike edge modes can lead to nonuniversal exponents in the current-voltage characteristic. PMID- 11864039 TI - Engineering electronic lifetimes in artificial atomic structures. AB - By means of atomic manipulation, 51 Ag atoms have been precisely positioned to form a triangle with a base length of 245 A on a Ag(111) substrate. The scattering of the surface electrons at these adatoms results in a complex interference pattern. Spectroscopic data and dI/dV maps taken inside the triangle have been quantitatively evaluated by multiple scattering calculations of the wave pattern. Adjustment of the scattering parameters to the data yields the properties of the scatterers and the electron lifetimes. The experimental results for the electron lifetimes deviate from a (E-E(F))(-2) dependence and reflect the electronic band structure at the surface as well as the local influence of the triangle. PMID- 11864040 TI - Electron scattering at steps: image-potential states on Cu(119). AB - The dynamics of image-potential states on Cu(119) have been studied with two photon photoemission. Direction-dependent quasielastic scattering processes with large momentum transfer are attributed to the finite terrace-width distribution on the stepped surface. This effectively couples image-potential states via interband scattering and leads to an asymmetry of the decay rate. Electrons in the first image-potential state live apparently longer when running upstairs. PMID- 11864041 TI - Real-space measurement of the potential distribution inside organic semiconductors. AB - We demonstrate that the soft nature of organic semiconductors can be exploited to directly measure the potential distribution inside such an organic layer by scanning-tunneling microscope (STM) based spectroscopy. Keeping the STM feedback system active while reducing the tip-sample bias forces the tip to penetrate the organic layer. From an analysis of the injection and bulk transport processes it follows that the tip height versus bias trace obtained in this way directly reflects the potential distribution in the organic layer. PMID- 11864042 TI - Kondo effect of single Co adatoms on Cu surfaces. AB - The Kondo resonance of Co adatoms on the Cu(100) and Cu(111) surfaces has been studied by scanning tunneling spectroscopy. We demonstrate the scaling of the Kondo temperature T(K) with the host electron density at the magnetic impurity. The quantitative analysis of the tunneling spectra reveals that the Kondo resonance is dominated by the Cu bulk electrons. While at the Cu(100) surface both tunneling into the hybridized localized state and into the substrate conduction band contribute to the Kondo resonance, the latter channel is found to be dominant for Cu(111). PMID- 11864043 TI - Flux flow of Abrikosov-Josephson vortices along grain boundaries in high temperature superconductors. AB - Low-angle grain boundaries (GBs) in superconductors exhibit intermediate Abrikosov vortices with Josephson cores, whose length l along GB is smaller than the London penetration depth, but larger than the coherence length. We found an exact solution for a periodic vortex structure moving along GBs in a magnetic field H and calculated the flux flow resistivity R(F)(H), and the nonlinear voltage-current characteristics. The predicted R(F)(H) dependence describes well our experimental data on 7 unirradiated and irradiated YBa(2)Cu(3)O(7) bicrystals, from which the core size l(T), and the intrinsic depairing density J(b)(T) on nanoscales of a few GB dislocations were measured for the first time. The observed J(b)(T) = J(b0)(1-T/T(c))(2) indicates a significant order parameter suppression on GB. PMID- 11864044 TI - Nanoscale one-dimensional scattering resonances in the CuO chains of YBa(2)Cu(3)O(6+x). AB - We present scanning tunneling spectroscopy measurements of the CuO chain plane in YBa(2)Cu(3)O(6+x), showing an approximately 25 meV gap in the local density of states (LDOS) filled by numerous intragap resonances: intense peaks in LDOS spectra associated with one-dimensional, Friedel-like oscillations. We discuss how these phenomena shed light on recent results from other probes, as well as their implications for phenomena in the superconducting CuO(2) plane. PMID- 11864045 TI - Impurity states and interlayer tunneling in high temperature superconductors. AB - We argue that the scanning tunneling microscope (STM) images of resonant states generated by doping Zn or Ni impurities into Cu-O planes of BSCCO are the result of quantum interference of the impurity signal coming from several distinct paths. The impurity image seen on the surface is greatly affected by interlayer tunneling matrix elements. We find that the optimal tunneling path between the STM tip and the metal (Cu, Zn, or Ni) d(x(2)-y(2)) orbitals in the Cu-O plane involves intermediate excited states. This tunneling path leads to the fourfold nonlocal filter of the impurity state in Cu-O plane that explains the experimental impurity spectra. Applications of the tunneling filter to the Cu vacancy defects and "direct" tunneling into Cu-O planes are also discussed. PMID- 11864046 TI - Charge and spin structure in of YBa(2)Cu(3)O(6.35). AB - Neutron scattering has been used to measure the charge and spin structure in the highly underdoped superconductor YBa(2)Cu(3)O(6.35). Incommensurate static charge ordering is found that remains at high temperatures. The magnetic pattern is complex with a resonance and incommensurate structure observed at low temperatures. The results clarify the role of striped phases in YBa(2)Cu(3)O(6+x) superconductors. PMID- 11864047 TI - Absence of a loss of in-plane infrared spectral weight in the pseudogap regime of Bi(2)Sr(2)CaCu(2)O(8+delta). AB - The ab-plane reflectance of Bi(2)Sr(2)CaCu(2)O(8+delta) (Bi-2212) thin films was measured in the 30-25 000 cm(-1) range for one underdoped ( T(c) = 70 K), and one overdoped sample ( T(c) = 63 K) down to 10 K. We find similar behaviors in the temperature dependence of the normal-state infrared response of both samples. Above T(c), the effective spectral weight, obtained from the integrated conductivity, does not decrease when T decreases, so that no opening of an optical pseudogap is seen. We suggest that these are consequences of the pseudogap opening in the k = (0,pi) direction and of the in-plane infrared conductivity being mostly sensitive to the k = (pi,pi) direction. PMID- 11864048 TI - Charge/orbital ordering structure of Pr(1-x)Ca(x)MnO(3)(x = 3/8) examined by low temperature transmission electron microscopy. AB - The structural phase transition of Pr(1-x)Ca(x)MnO(3)(x = 3/8) was investigated by means of low-temperature transmission electron microscopy. Superlattice reflection spots with a modulation wave vector q(1) = (0,1/2,0) appeared below 230 K, indicating formation of the d(3x(2-r(2))/d(3y(2)-r(2)) type of charge/orbital ordering. Below 150 K, a new series of superlattice reflection spots with a modulation wave vector q(2) = (1/4,1/4,1/2) appeared, suggesting an additional ordering of excess 1/8 Mn(3+), necessary due to the deviation of x from 1/2, with the occupation of the d(3z(2-r(2)) type of e(g) orbital. PMID- 11864049 TI - Surface chemistry of silicon nanoclusters. AB - We employ density functional and quantum Monte Carlo calculations to show that significant changes occur in the gap of fully hydrogenated nanoclusters when the surface contains passivants other than hydrogen, in particular atomic oxygen. In the case of oxygen, the gap reduction computed as a function of the nanocluster size provides a consistent interpretation of several recent experiments. Furthermore, we predict that other double bonded groups also significantly affect the optical gap, while single bonded groups have a minimal influence. PMID- 11864050 TI - Imaging the local density of states of optical corrals. AB - This paper reports the experimental observation, at optical frequencies, of the electromagnetic local density of states established by nanostructures corresponding to the recently introduced concept of optical corral [G. Colas des Francs et al., Phys. Rev. Lett. 86, 4950 (2001)]. The images obtained by a scanning near-field optical microscope under specific operational conditions are found in agreement with the theoretical maps of the optical local density of states. A clear functionality of detection by the scanning near-field optical microscope is thereby identified since the theoretical maps are computed without including any specific tip model. PMID- 11864051 TI - Antiferroelectric behavior in symmetric KNbO(3)/KTaO(3) superlattices. AB - Antiferroelectric behavior is observed in artificially layered KTaO (3)/KNbO (3) perovskite superlattices. While KTaO (3) and KNbO (3) are ferroelectric and paraelectric, respectively, the superlattice appears antiferroelectric based on an increase in the dielectric constant with applied dc bias. This dielectric behavior is inconsistent with the nonlinear response for either paraelectric or ferroelectric materials. However, an increase in the dielectric constant with applied electric field is consistent with antiferroelectric behavior. The antiferroelectric ordering appears to be induced by cation modulation imposed by the superlattice. PMID- 11864052 TI - Spin-lattice relaxation enhancement in liquid gallium confined within nanoporous matrices. AB - Nuclear spin relaxation for liquid gallium embedded into nanoporous matrices was found to accelerate remarkably compared to the bulk melt. NMR measurements on two gallium isotopes showed that the dominant mechanism of relaxation was changed from magnetic to quadrupolar and the relation rate depended on the Larmor frequency. The correlation time of electric field gradient fluctuations was estimated using data for quadrupolar relaxation contribution and was found to increase drastically compared to bulk, which corresponded to slowing down mobility in confined liquid gallium. PMID- 11864053 TI - Surface charging and impulsive ion ejection during ultrashort pulsed laser ablation. AB - We report time-resolved studies using femtosecond laser pulses, accompanied by model calculations, that illuminate the difference in the dynamics of ultrashort pulsed laser ablation of different materials. Dielectrics are strongly charged at the surface on the femtosecond time scale and undergo an impulsive Coulomb explosion. This is not seen from metals and semiconductors where the surface charge is effectively quenched. PMID- 11864055 TI - Experimental simulation of two-particle quantum entanglement using classical fields. AB - We experimentally demonstrate simulation of two entangled quantum bits using classical fields of two frequencies and two polarizations. Multiplication of optical heterodyne beat signals from two spatially separated regions simulates coincidence detection of two particles. The product signal so obtained contains several frequency components, one of which can be selected by bandpass frequency filtering. The bandpassed signal contains two indistinguishable, interfering contributions, permitting simulation of four polarization-entangled Bell-like states. These classical field methods may be useful in small scale simulations of quantum logic operations that require multiparticle entanglement without collapse. PMID- 11864054 TI - Optimizing the encounter rate in biological interactions: Levy versus Brownian strategies. AB - An important application involving two-species reaction-diffusion systems relates to the problem of finding the best statistical strategy for optimizing the encounter rate between organisms. We investigate the general problem of how the encounter rate depends on whether organisms move in Levy or Brownian random walks. By simulating a limiting generalized searcher-target model (e.g., predator prey, mating partner, pollinator-flower), we find that Levy walks confer a significant advantage for increasing encounter rates when the searcher is larger or moves rapidly relative to the target, and when the target density is low. PMID- 11864056 TI - Communication with dynamically fluctuating states of light polarization. AB - The fast, irregular, fluctuations of the state of polarization of light output from an erbium doper fiber ring laser are used to communicate digital information. The experiments illustrate the application of a dynamical encoding and information recovery scheme that is robust to perturbations of the communication channel, a standard single mode fiber. A fiber-optic polarization analyzer was used to measure and visualize the polarization dynamics at nanosecond time scales on the Poincare sphere. PMID- 11864057 TI - Efficient classical simulation of continuous variable quantum information processes. AB - We obtain sufficient conditions for the efficient simulation of a continuous variable quantum algorithm or process on a classical computer. The resulting theorem is an extension of the Gottesman-Knill theorem to continuous variable quantum information. For a collection of harmonic oscillators, any quantum process that begins with unentangled Gaussian states, performs only transformations generated by Hamiltonians that are quadratic in the canonical operators, and involves only measurements of canonical operators (including finite losses) and suitable operations conditioned on these measurements can be simulated efficiently on a classical computer. PMID- 11864058 TI - Thermalizing quantum machines: dissipation and entanglement. AB - We study the relaxation of a quantum system towards the thermal equilibrium using tools developed within the context of quantum information theory. We consider a model in which the system is a qubit, and reaches equilibrium after several successive two-qubit interactions (thermalizing machines) with qubits of a reservoir. We characterize completely the family of thermalizing machines. The model shows a tight link between dissipation, fluctuations, and the maximal entanglement that can be generated by the machines. The interplay of quantum and classical information processes that give rise to practical irreversibility is discussed. PMID- 11864059 TI - Characterizing the structure of small-world networks. AB - We give exact relations for small-world networks (SWN's) which are independent of the "degree distribution," i.e., the distribution of nearest-neighbor connections. For the original SWN model, we illustrate how these exact relations can be used to obtain approximations for the corresponding basic probability distribution. In the limit of large system sizes and small disorder, we use numerical studies to obtain a functional fit for this distribution. Finally, we obtain the scaling properties for the mean-square displacement of a random walker, which are determined by the scaling behavior of the underlying SWN. PMID- 11864061 TI - Comparative simulation study of colloidal gels and glasses. AB - Using computer simulations, we identify the mechanisms causing aggregation and structural arrest of colloidal suspensions interacting with a short-ranged attraction at moderate and high densities. Two different nonergodicity transitions are observed. As the density is increased, a glass transition takes place, driven by excluded volume effects. In contrast, at moderate densities, gelation is approached as the strength of the attraction increases. At high density and interaction strength, both transitions merge, and a logarithmic decay in the correlation function is observed. All of these features are correctly predicted by mode coupling theory. PMID- 11864060 TI - Effect of solvation on hole motion in DNA. AB - An excess charge on a DNA chain in solution interacts with polar solvent molecules and mobile counterions. We give the first theoretical estimate of the resulting hole self-localization energy and calculate the corresponding contribution to hole mobility on a DNA stack consisting of a single base pair repeated. PMID- 11864062 TI - Cabello's nonlocality and linear optics. PMID- 11864063 TI - Rigorous bounds to retarded learning. PMID- 11864064 TI - Ligands of neuronal nicotinic acetylcholine receptor (nAChR): inferences from the Hansch and 3-D quantitative structure-activity relationship (QSAR) Models. AB - Neuronal acetylcholine ion channel receptors (nAChRs), that exist in several subtypes resulting from a different organisation of various subunits around the central ion channel, are involved in a variety of functions and disorders of the central nervous system. There is evidence to implicate a deficit of nAChRs in the symptomatology of severe neurologic pathologies, such as Alzheimer s and Parkinson s diseases. Reliable three-dimensional structures of nAChRs are not available yet, and this hampers adopting structure-based approaches in designing new ligands. Also pharmacophore models are not reliable enough to be used in ligand-based approaches to drug design and little structure-activity work has been reported so far. This paper deals with structure-activity relationships of a wide series of nicotinic ligands. It provides results from a study of the quantitative structure activity relationships (QSARs) based on literature data of about 270 nicotinic agonists, belonging to various chemical classes. The QSAR study was carried out by using either a classical Hansch approach or a Comparative Molecular Field Analysis (CoMFA). Within each congeneric series, Hansch-type equations revealed detrimental steric effects as the factors mainly modulating the receptor affinity, whereas CoMFA allowed us to merge progressively models obtained for each class of congeners into a more general one that showed good cross-validation statistics. The CoMFA coefficient isocontour maps illustrated, at the 3-D level, the most relevant interactions responsible for a high receptor affinity, whereas the robustness of the global three-dimensional QSAR/CoMFA (n = 206, q(2) = 0.749, r(2) = 0.847, s= 0.600) model was supported by the high value of the prediction statistics (r(2)pred = 0.961) and confirmed by the satisfactory predictions of the affinity data of an external set of 18 recently published ligands with chemical structures even quite diverse from those included in the training set. PMID- 11864065 TI - Nitric-oxide releasing molecules: a new class of drugs with several major indications. AB - Nitric oxide (NO) deficiency has been implicated in many pathological and physiological processes within the mammalian body providing a plausible biologic basis for the use of NO replacement therapy in these conditions. Exogenous NO sources may hopefully constitute a powerful way to supplement NO when the body cannot generate enough for normal biological functions. This theory has opened up the possibility of designing new drugs that are capable of delivering NO into tissues and the bloodstream in a sustained and controlled manner. This objective has been reached by grafting an organic nitrate structure onto existing molecules with various spacers such as aliphatic or aromatic chain, with different degree of complexity. This approach has led to the synthesis of several new chemical entities in various pharmacological classes, whose profile seems to challenge the parent drug not only on the basis of new pharmacological properties but also on a better toxicological and safety profile. In this article, general aspects on NO and NO donors are reviewed. Major focus is placed upon recent developments of novel NO donors, NO releasing device(s) as well as innovative improvements to conventional NO donors. Several examples are given in some important therapeutic indications such as cardiovascular diseases (NO-aspirin), pain and inflammation (NO-paracetamol), osteoporosis and urinary incontinence (NO flurbiprofen with aliphatic spacer), Alzheimer s disease (NO-flurbiprofen with anti-oxidant spacer), respiratory disorders (NO-steroids). PMID- 11864067 TI - Living in the 'post-antibiotic era': could the use of probiotics be an effective strategy? PMID- 11864066 TI - 8th European Congress of Clinical Microbiology and Infectious Diseases Lausanne, Switzerland, May 25--28, 1997. PMID- 11864068 TI - Scedosporium species: the rising importance of newly emerging fungal pathogens. PMID- 11864069 TI - Controlling glycopeptide-resistant enterococci. PMID- 11864070 TI - Rhodococcus equi infection in non-HIV-infected patients. Two case reports and review. AB - OBJECTIVE: To review two recent cases in HIV-negative subjects in the light of literature reports (52 patients without HIV infection till 1994). METHODS: Epidemiology (animal contacts, risk factors, year, country), clinical presentation, diagnostic methods (X-ray, tomography, microbiological techniques), therapeutic approach (antibiotics, surgery) and outcome were evaluated on the basis of clinical literature reports. RESULTS: Tumors constituted an important predisposing factor and less frequently hepatobiliary pathology, rheumatologic diseases, iatrogenic causes, psychiatric pathology and trauma. Exposure to animals was reported by 55% of the patients. Pneumonia and pleurisy, without preferential localization, were detected in 50% of the patients. Etiologic diagnosis was usually obtained after an invasive collection. Combined medical therapy and surgery were required by 27.8% of the patients, and 16.7% of the patients died. CONCLUSIONS: In recent years the number of Rhodococcus equi cases has been rising also in HIV-negative patients. The infection is ubiquitous. Accurate diagnosis and the prompt selection of the most appropriate therapy depend on close cooperation between clinicians and microbiologists. PMID- 11864071 TI - Vaginal pH neutralization by semen as a cofactor of HIV transmission. AB - OBJECTIVE: To explain the high frequency of heterosexual transmission of AIDS in subSaharan countries, by investigating post-intercourse semen retention in the vagina for local pH neutralization as a possible interference factor with acidic inactivation of HIV virions. METHODS: Two semen markers, prostate-specific antigen and prostatic acid phosphatase, were measured and compared with the pH values in the vaginal washes of 69 women from the Central African Republic. The capacity of semen to raise the vaginal pH was also investigated in vitro. RESULTS: Of 61 non-menstrual specimens, 74% contained at least one semen marker. The specimens with high levels of markers (group I) displayed an almost neutral pH (median 6.1), at variance with the semen-free group II (median: 3.7, P<0.003), and with group III (median: 4.0) corresponding to low or past semen retention. The in vitro study confirmed the high neutralization capacity of semen. CONCLUSIONS: It is expected that post-intercourse neutralization of pH will both favor male-to-female transmission and prevent the acidity-associated loss of infectivity of the female-derived virions, thus allowing female-to-male transmission during further sexual contact. PMID- 11864072 TI - Is TNF-alpha a prognostic factor in patients with sepsis? AB - OBJECTIVE: To determine tumor necrosis factor-alpha (TNF-alpha) levels in a prospective study in 58 hospitalized patients in a department of internal medicine (63 episodes, 29 in immunocompromised patients) during a 7-month period. METHODS: Patients fulfilling the following criteria were included: clinical evidence of acute infection, temperature >38.2 degrees C, tachycardia >90 beats/min, tachypnea >20 breaths/min. Samples were taken from day 1 up to day 13 after an infection was diagnosed, and TNF-alpha was determined by enzyme immunoassay. RESULTS: In 29 episodes (46.0%) the infection was microbiologically documented. The median of the TNF-alpha levels in the Gram-negative episodes was significantly higher than that in the Gram-positive episodes (p=0.002). Thirteen of 63 episodes (20.6%) had a fatal outcome. With respect to all measured values, the non-survivors had a significantly higher median of TNF-alpha levels than the survivors (p=0.0001). There was, however, great interpatient and intrapatient variability in TNF-alpha levels; thus, no unequivocal correlation between TNF alpha and outcome could be documented. CONCLUSIONS: Our data indicate that the influence of the infecting organism on TNF-alpha kinetics is less pronounced than that of the underlying disease. PMID- 11864073 TI - Retrospective study of outcome in patients treated for Staphylococcus aureus bacteremia. AB - OBJECTIVE: To investigate whether a change in current treatment practice for Staphylococcus aureus bacteremia from flucloxacillin and aminoglycoside to flucloxacillin and fusidic acid was associated with any changes in outcome. METHODS: A retrospective analysis was carried out of 316 episodes of S. aureus bacteremia diagnosed and treated in a tertiary hospital complex between 1983 and 1993. Outcomes considered were (1) death related to the infection and (2) relapse following cessation of antibiotic therapy. RESULTS: Mortality related to infection, which occurred in 24% of patients, was unrelated to treatment with the combination of flucloxacillin and fusidic acid; however, increasing age was a significant risk factor (OR per decade=1.35, 95% CI=1.18-1.55), and increasing duration of treatment (OR per week of treatment=0.63, 95% CI=0.52-0.77), use of flucloxacillin (OR=0.30, 95% CI=0.14-0.64), presence of an intravascular device (OR=0.39, 95% CI=0.20-0.78) and presence of a skin lesion (OR=0.51, 95% CI=0.26 0.99) were significant protective factors. The only factor significantly related to relapse, which occurred in 11% of patients, was treatment with the combination of flucloxacillin and fusidic acid (OR=0.32, 95% CI=0.12-0.85). There was approximately a 70% reduction in the risk of relapse if this combination was used. CONCLUSIONS: This retrospective analysis suggests a clinically important protective effect of fusidic acid against relapse in patients with S. aureus bacteremia. Although the results were adjusted for potential confounding factors, the possibility of bias remains. There is a need for a prospective randomized trial to evaluate the effectiveness of flucloxacillin and fusidic acid for treating S. aureus bacteremia. PMID- 11864074 TI - Early Lyme disease: Humoral immune status and treatment. AB - OBJECTIVE: To investigate the humoral immune status and the effect of antibiotic treatment in Bulgarian patients with early Lyme disease. METHODS: A total of 34 early Lyme disease patients was examined, 16 with erythema migrans and 18 with non-specific systemic symptoms. Serum samples from all patients and from 12 healthy controls were tested for total immunoglobulins (IgG, IgA and IgM), hemolytic activity of complement (CH50) and immune complexes (ICs). The patients were treated with doxycycline (100 mg orally, twice daily for 10 to 15 days, in one or two courses). RESULTS: The patients showed significantly increased IC levels (P<0.01 for patients with erythema migrans and P<0.001 for patients with non-specific symptoms). There were no significant changes in the levels of total hemolytic complement and total immunoglobulins (IgG, IgA and IgM). The clinical outcome was satisfactory in 21 of the 34 patients (61.8%) after treatment with doxycycline for 10 to 15 days. The rest of the patients (38.2%) failed to respond to the therapy, and continued to report various complaints, such as arthralgia, myalgia, paresthesia, headache, fatigue or recurrent rash. All of these had elevated levels of IC. After a second course of treatment with the same antibiotic regimen these patients had resolution of symptoms (12 patients) or improvement (1 patient). CONCLUSIONS: Immunologic investigation may be useful in determining treatment strategy in Lyme disease. Elevated IC levels may indicate a need for more prolonged antibiotic therapy. PMID- 11864075 TI - Evaluation of clonal relatedness of extended-spectrum beta-lactamase-producing Proteus mirabilis isolates by quantitative antibiogram and RAPD typing. AB - OBJECTIVE: To delineate, using two different typing systems, the clonal relatedness of 40 isolates of extended-spectrum beta-lactamase (ESbetaIa) producing Proteus mirabilis obtained over a period of 7 years in six hospitals in the Paris area and two in Pas-de-Calais. METHODS: Random amplified polymorphic DNA (RAPD) polymerase chain reaction typing was applied by using three random primers on the ESbetaIa-producing P. mirabilis isolates and on isogenic Escherichia coli strains with or without plasmids encoding the representative resistance pattern transferred from P. mirabilis. Quantitative antibiogram typing, which was also applied to the P. mirabilis isolates, was used to define the euclidean distance between these strains. RESULTS: After having demonstrated that P. mirabilis plasmids did not influence chromosomal DNA amplification, we could classify the ESbetaIa-producing P. mirabilis isolates into 12 groups based on RAPD fingerprints. The same isolates were classified into 19 groups by quantitative antibiogram typing. Despite this difference in group numbers, general concordance between the typing systems was observed. This allowed us to show that the greater number of isolates in some hospitals belonged to a single strain and that single isolates obtained in different hospitals generally represented unique strains. CONCLUSIONS: A small number of ESbetaIa-producing P. mirabilis strains was isolated during 7 years in the eight medical centers studied, and the number of different strains identified suggested that inter hospital transfer had not occurred. PMID- 11864076 TI - Differentiation and susceptibility of Citrobacter isolates from patients in a university hospital. AB - OBJECTIVE: Recently a publication of Brenner et al. introduced 11 genetically distinct species within the genus Citrobacter. These newly recognized Citrobacter species can be classified by means of their biochemical characteristics. The aim of this study was to examine the distribution and susceptibility of Citrobacter isolates in our patient population. METHODS: A total of 126 samples---containing a Citrobacter species---was collected from 116 hospitalized patients during a 6 month period. Organisms were identified according to standard procedures. Antimicrobial susceptibility testing was performed by agar dilution on Mueller Hinton agar, and interpretation was based on NCCLS criteria. RESULTS: C. freundii was the most common organism isolated (n=59), followed by C. braakii (n=25) and C. koseri (n=23). The urinary tract and the respiratory tract were found to be the predominant sites of colonization or infection, accounting for 45% and 32% of all isolates respectively. It appeared that young children (<12 months old) and the elderly were most at risk of acquiring Citrobacter. Two-thirds of all specimens contained other organisms in addition to Citrobacter. Most Citrobacter isolates were related with a predisposing factor. Species-related differences were found in the susceptibility pattern. CONCLUSIONS: These findings suggest that citrobacteria are important opportunistic pathogens contributing to colonization or infection in our hospital population. PMID- 11864077 TI - Susceptibility of Streptococcus pyogenes from throat cultures to macrolide antibiotics and influence of collection criteria. AB - OBJECTIVE: To assess the incidence of resistance to erythromycin and to the three other macrolide antibiotics most extensively used in Italy (azithromycin, clarithromycin and roxithromycin) among clinical strains of Streptococcus pyogenes freshly isolated from throat cultures of pediatric patients in an area of Central Italy. METHODS: Two sets of isolates were examined. The strains of the first set (n=100) were collected according to a protocol admitting only throat swabs from untreated patients with symptoms of acute pharyngotonsillitis. The second set (n=180) consisted of strains isolated from throat cultures during the routine activity of diagnostic laboratories, no particular protocol being applied. RESULTS: A trimodal distribution of strains was observed in relation to their macrolide susceptibility levels: two clusters were constituted by highly susceptible and highly resistant strains, respectively; a third, middle cluster consisted of strains displaying low-level resistance (or even intermediate susceptibility, in a minority of isolates, to clarithromycin). The distribution of individual isolates in the three modal clusters was the same with all four drugs. Both MIC ranges and MIC50s almost overlapped in the isolates of the two sets, whereas MIC90s were far higher in the strains of the second set (4 micro g/mL for clarithromycin, 8 micro g/mL for erythromycin and azythromycin, and 16 micro g/mL for roxithromycin) than in those of the first (0.125 micro g/mL for all four drugs). Resistant strains were 5% among the isolates of the first set and three times as many among those of the second. CONCLUSIONS: The lower incidence of macrolide resistance recorded in the first set is probably more reliable: the threefold incidence observed in the second set may be overestimated due to the lower frequency of strains involved in drug-responsive infections and to the increased occurrence of strains from unsuccessfully treated patients. PMID- 11864078 TI - Pharmacokinetics of 18F-labeled trovafloxacin in normal and Escherichia coli infected rats and rabbits studied with positron emission tomography. AB - OBJECTIVE: To measure tissue pharmacokinetics of trovafloxacin (CP 99,219) in normal and infected animals by both direct tissue radioactivity measurements and positron emission tomography (PET). METHODS: Concentrations of [18F]trovafloxacin were measured in normal and infected rats (n=6/group), at 10, 30, 60, and 120 min after injection, by radioactivity measurements. In normal rabbits (n=4) and rabbits with Escherichia coli thigh infection (n=4), tissue concentrations of drug were measured over 2 h with PET. After acquiring the final images, the rabbits were killed and tissue concentrations measured with PET were compared to the results of direct tissue radioactivity measurements. RESULTS: In both species, there was rapid distribution of [18F] trovafloxacin in most peripheral organs. Peak concentrations of more than five times the MIC90 of most Enterobacteriaceae and anaerobes (>100-fold for most organisms) were achieved in all tissues and remained above this level for >2 h. Particularly high peak concentrations were achieved in the kidney (>75 micro g/g), liver (>100 micro g/g), blood (>40 micro g/g), and lung (>10 micro g/g). Even though the concentration of trovafloxacin in infected muscle was reduced (p<0.01), the peak concentration was still >4 micro g/g and tissue levels remained above 2 micro g/g for more than 2 h. Due to the lower concentrations that were achieved in the brain (peak approximately 5 micro g/g), it is expected that trovafloxacin will have limited central nervous system toxicity. CONCLUSION: PET with [18F]trovafloxacin is a useful technique for non-invasive measurements of tissue pharmacokinetics. PMID- 11864079 TI - Comparative in vitro pharmacodynamics of BO-2727, meropenem and imipenem against Gram-positive and Gram-negative bacteria. AB - OBJECTIVE: To investigate and compare the in vitro pharmacodynamics of three carbapenems: imipenem, meropenem and BO-2727. METHODS: The following studies were performed: (1) comparative studies of the rate of killing of the three carbapenems of reference strains of Gram-positive and Gram-negative bacteria at a concentration corresponding to the 1-h serum level following 500 mg intravenously in humans; (2) comparative studies of the rate of killing of BO-2727, meropenem and imipenem at different antibiotic concentrations of reference strains of Gram positive and Gram-negative bacteria; (3) comparative studies of the rate of killing of BO-2727, meropenem and imipenem of bacteria which are phenotypically tolerant; (4) studies of the postantibiotic effect of BO-2727 using viable counts and optical density; (5) studies of the postantibiotic sub-MIC effect (PA SME) of BO-2727 using optical density. RESULTS: No difference in killing rate was noted between the three carbapenems, and there was no concentration-dependent killing of the Gram-negative strains after 6 h. A pronounced paradoxical effect was seen against Staphylococcus aureus. All three antibiotics were able to kill phenotypically tolerant bacteria. Only very short or no postantibiotic effect of BO-2727 was found against the investigated strains. Very long PA SMEs were noted for the Gram-negative strains, although there was a pronounced variation for the different strains of Pseudomonas aeruginosa. CONCLUSION: There was no significant difference between the studied carbapenems in their pharmacodynamic properties. All three antibiotics acted similarly to other beta-lactam antibiotics. PMID- 11864080 TI - Postantibiotic effects with Bacteroides fragilis determined by viable counts and CO2 generation. AB - OBJECTIVE: To study the postantibiotic effect (PAE) for Bacteroides fragilis after exposure to common anaerobic antimicrobials with two different methods, by viable counting and by measuring CO2 generation in a BACTEC(R) blood culture system. METHODS: Four strains of B. fragilis were exposed for 1, 2 and 4 h to cefoxitin, chloramphenicol, clindamycin, imipenem or metronidazole at concentrations from 1 to 16 x MIC. The drugs were removed by dilution into BACTEC 7A(R) vials and growth determined with viability counts and CO2 production. RESULTS: The durations of the PAEs determined by the two methods correlated well (r=0.913, p<0.005). PAEs of up to 4-5 h were induced by imipenem and metronidazole with achievable concentrations and exposure durations. Chloramphenicol induced short or no PAEs, but cefoxitin and clindamycin induced PAEs up to 2 h with high AUC values. The imipenem PAEs and the short cefoxitin and clindamycin PAEs were dependent on AUC. CONCLUSIONS: Significant PAEs against B. fragilis were induced by imipenem and metronidazole. Determining PAE by measuring CO2 production is an accurate and less time-consuming alternative to the conventional method of viable counts. PMID- 11864081 TI - Effect of clindamycin in a model of acute murine toxoplasmosis. AB - OBJECTIVE: To characterize the antitoxoplasma activity of clindamycin in a murine model of acute toxoplasmosis. METHODS: Rates of survival and mean survival times of Swiss Webster mice infected intraperitoneally with 106-102 tachyzoites of the RH strain of Toxoplasma gondii treated with clindamycin or sulfamethoxazole (positive control) or untreated (negative control) were compared. Survivors were submitted to examination of untreated brain tissue preparations, intraperitoneal and peroral subinoculations of brain tissue homogenates into fresh mice, and to pathohistology, including immunohistochemistry, of brain and lungs. RESULTS: The effect of clindamycin treatment (400 mg/kg/day) on infected Swiss Webster mice was inoculum size dependent, ranging from no survivals in animals infected with 106 parasites, to 100% survivals with an inoculum of 102. Treatment initiated 24 h before and at time of infection prolonged mean survival times comparably to sulfamethoxazole, and significantly when compared to untreated controls. In contrast, treatment initiated 48 h postinfection with an inoculum of 106 did not postpone death. In the clindamycin-treated survivors, there was no biological or histologic evidence for the persistence of toxoplasma. CONCLUSIONS: The results obtained show that at an appropriate parasite dose/drug dose ratio, clindamycin is strongly toxoplasmacidal in a murine model of acute toxoplasmosis. PMID- 11864082 TI - A nested polymerase chain reaction for detection of Legionella pneumophila in clinical specimens. AB - OBJECTIVE: Because presently used methods for diagnosis of Legionella pneumonia lack sufficient sensitivity and sometimes specificity and rapidity, the detection of Legionella spp. by amplification of nucleic acids might be valuable. However, performing polymerase chain reaction (PCR) on clinical samples such as sputum is difficult because of the presence of extraneous DNA and inhibitors of the reaction. An attempt to circumvent these problems was made. METHODS: A nested PCR method was devised using primers from the mip gene of Legionella pneumophila. This PCR was tested on pure cultures of legionellae and clinical isolates of other bacteria. Clinical samples (bronchoalveolar lavage fluid, bronchial aspirate and sputum) from patients who suffered from legionellosis and samples from patients who suffered from other causes of pneumonia were also tested. RESULTS: The PCR was specific for L. pneumophila and no non-Legionella bacteria reacted. Ten to 50 colony forming units of Legionella in the sample could be detected. Twenty-two of 25 clinical samples were positive among patients suffering from pneumonia proven to be due to L. pneumophila serogroups 1, 3, 4, 5 and 6. Two of the three negative samples were from patients who had been treated with adequate therapy for at least 2 days and were culture negative. However, nine other culture-negative samples were PCR positive, of which seven came from patients who had been treated for 3-7 days. All pneumonia patients in the control group proved negative in PCR. A commercial kit for DNA preparation from clinical samples, based on absorption of nucleic acids to silica gel, was superior to the traditional phenol/chloroform extraction and increased the rapidity, simplicity and sensitivity of the procedure. CONCLUSIONS: A nested, simplified and rapid PCR method using mip primers proved to be more sensitive than culture and as sensitive and specific as other PCR procedures previously reported. PMID- 11864083 TI - PCR ribotyping and arbitrarily primed PCR for the comparison of enterotoxigenic Bacteroides fragilis strains from two Polish university hospitals. AB - OBJECTIVE: To study the clinical incidence and possible clonal relatedness of enterotoxigenic strains of Bacteroides fragilis among pediatric and adult patients in two Polish university hospitals. METHODS: Fecal samples from 201 adults and 131 infants (with or without diarrhea) and vaginal samples from 100 pregnant women nursed in two Polish university hospitals were analyzed with respect to carriage of enterotoxin-producing Bacteroides fragilis (ETBF). This putative pathogen was identified by cultivation and subsequent cytopathogenicity testing of culture supernatants on HT/29 C1 cells. RESULTS AND DISCUSSION: Two ETBF strains were isolated from childrens' feces; two additional strains were isolated from adults, and from the vaginal samples only a single strain was isolated. One strain (W2) was isolated from a child with diarrhea. These incidence figures, the fact that all ETBF isolates were shown to produce strongly differing amounts of the cytotoxin, and the genetic unrelatedness of the strains as demonstrated by two different PCR-mediated DNA typing procedures, indicates that clonal spread of ETBF is presently not a clinical problem in these hospitals. It was shown that PCR-mediated ribotyping and arbitrarily primed PCR can be applied with success to study the epidemiology of ETBF. PMID- 11864084 TI - Detection of Borrelia afzelii, Borrelia burgdorferi sensu stricto, Borrelia garinii and group VS116 by PCR in skin biopsies of patients with erythema migrans and acrodermatitis chronica atrophicans. AB - OBJECTIVE: To evaluate the diagnostic performance of two polymerase chain reaction (PCR) procedures using skin biopsies of 20 erythema migrans (EM) and 24 acrodermatitis chronica atrophicans (ACA) patients. METHODS: One assay amplified a fragment of the outer surface protein (Osp) A gene. The second method amplified the spacer region between the 5S and 23S rRNA genes; hybridization of this fragment allowed identification of Borrelia burgdorferi sensu lato species. RESULTS: Among EM patients, both assays detected Borrelia DNA in 15 samples. Among ACA patients, the ospA PCR detected 15 positives and 10 samples were positive by 5S-23S PCR. In 19 samples one species was detected, 15 skin biopsies contained Borrelia afzelii, and Borrelia garinii was found in two patients. Group VS116 was detected in two EM patients, and therefore this group has pathogenic potential. Mixed infections of B. afzelii and B. garinii, group VS116 or B. burgdorferi sensu stricto were found in three EM and three ACA patients. CONCLUSIONS: Diagnosis of EM and ACA by PCR is useful and knowledge of the presence of species may be used to predict the course of disease or the need for further antibiotics. PMID- 11864085 TI - Direct detection of verotoxin genes in stool samples by polymerase chain reaction in hemolytic uremic syndrome patients in France. AB - OBJECTIVE: To reassess the occurrence of verocytotoxin-producing Escherichia coli (VTEC) in French hemolytic uremic syndrome (HUS) patients. METHODS: From March 1991 to January 1995, direct detection of verotoxin genes (VT) by the polymerase chain reaction (PCR) was performed on stool samples from 169 patients suffering from HUS. RESULTS: Fifty-one were PCR positive (30.1%); one was positive for the VT1 gene and the others for the VT2 gene. VTEC was isolated from only 32 of the 51 PCR-positive samples. E. coli O157:H7 was isolated from five patients. E. coli O111 was isolated from seven patients during an outbreak of HUS. Among the other VT2 E. coli strains, only four were serotypable. Of the 51 PCR-positive stools, 19 were culture negative for VTEC. CONCLUSIONS: This study provides evidence that in France E. coli O157 and other VTEC serotypes are involved in HUS. PMID- 11864086 TI - Detection and subgrouping of respiratory syncytial virus directly from nasopharyngeal aspirates. AB - OBJECTIVE: To develop a reverse transcription-polymerase chain reaction (RT-PCR) based assay to identify the subgroup of the infecting respiratory syncytial virus (RSV) strain directly from nasopharyngeal aspirates (NPAs). METHODS: A total of 154 NPAs which were positive for RSV antigen by direct immunofluorescence were subjected to RT-PCR. The primers used were designed to give products for subgroup A and B which were of different sizes and easily visualized on agarose electrophoresis. The PCR products were further analyzed by restriction analysis using enzymes which were unique or rare cutters within the PCR amplimer. RESULTS: It was possible to confirm RSV infection in 70% of the NPA samples studied. Of these, 92.6% belonged to the A group, and only 7.4% to the B group. Within the A group, six subgroups were identified using restriction analysis, while all B group samples were identical to the prototype B strain, 18537. CONCLUSION: RT-PCR performed on RNA isolated directly from NPAs provides a quick, easy-to-use, reasonably sensitive method to identify and group the infecting RSV strain. PMID- 11864088 TI - Circumventing antibiotic resistance in specialized hospital units. PMID- 11864087 TI - Oral supplementation with lactic acid-producing bacteria during intake of clindamycin. AB - OBJECTIVE: To study the effect of administration of clindamycin with or without supplementation of the intestinal microflora with Bifidobacterium bifidum and Lactobacillus acidophilus. METHODS: Twenty-three healthy subjects received clindamycin by mouth for 7 days. Eleven of the subjects also received capsules containing lyophilized L. acidophilus and B. bifidum for 14 days. The other 12 subjects received placebo. RESULTS: There was a marked decrease in total numbers of anaerobic bacteria during the administration of clindamycin. In the lactic acid bacteria-supplemented group, a tendency towards delayed reduction and earlier increase in bifidobacteria was observed, and two of 11 subjects (18%) were colonized with Clostridium difficile, in comparison with five of 12 (41%) in the placebo group. The total number of microorganisms was significantly higher in the lactic acid bacteria-supplemented group than in the placebo group (p=0.02) 4 days after the end of clindamycin administration. The difference was mainly due to higher counts of Escherichia coli and enterococci. Mean levels of other enterobacteria increased less in the lactic acid bacteria-supplemented group than in the placebo group between days 0 and 14. CONCLUSIONS: The recolonization with aerobic and anaerobic microorganisms was faster in the lactic acid bacteria supplemented group than in the placebo group. This may be of importance in preventing colonization with C. difficile. PMID- 11864089 TI - Incidence of Bilophila wadsworthia in appendiceal, peritoneal and fecal samples from children. PMID- 11864090 TI - Three-hour blood culture detection of Streptococcus pneumoniae. PMID- 11864091 TI - Native valve endocarditis with persistent embolization due to Neisseria mucosa. PMID- 11864092 TI - Quadri-valvular endocarditis caused by Streptococcus mutans. PMID- 11864093 TI - Secondary prophylaxis of Rhodococcus equi pneumonia in HIV infection: breakthrough despite rifampicin/erythromycin suppressive therapy. PMID- 11864094 TI - Intravenous penetration of fluconazole during endophthalmitis. PMID- 11864095 TI - Comparison of two DNA extraction methods on inhibitory sputum samples. PMID- 11864096 TI - Rapid detection of toxigenic Clostridium difficile strains by a nested PCR of the toxin B gene. PMID- 11864098 TI - Some reflections on the possible link between bovine spongiform encephalopathy and the new variant of Creutzfeldt---Jakob disease. PMID- 11864097 TI - Selection of rifampicin-resistant Mycobacterium tuberculosis does not occur in the presence of low concentrations of rifaximin. PMID- 11864099 TI - Tick-borne zoonoses in Europe. PMID- 11864100 TI - Pharmacokinetics of cefetamet pivoxil and interaction with cisapride and N acetylcysteine. AB - OBJECTIVE: To evaluate the pharmacokinetics of cefetamet pivoxil and possible interaction with N-acetylcysteine and cisapride in healthy volunteers. METHODS: In a double-blind, randomized three-way crossover study with 12 healthy male volunteers, serum and urine concentrations of cefetamet were determined over 12 h by a validated bioassay method after oral administration of 0.5 g cefetamet pivoxil and, randomly, placebo, 5x20 mg cisapride, or 0.6 g N-acetylcysteine. RESULTS: The study medications were well tolerated, although there were 10 cases of altered bowel movements, two cases of mild, transient headache and one case of increased serum transferase levels (AST and ALT). The mean peak serum level of cefetamet pivoxil in the placebo group was 4.86plus minus1.35 mg/L. The urine recovery/24 h in the placebo group was 41.9plus minus3.8% of the oral dose. The elimination half-life was 3.56plus minus0.92 h. N-Acetylcysteine had no effect on the pharmacokinetics of cefetamet pivoxil. With concomitant administration of cisapride there was an accelerated absorption of cefetamet pivoxil and a slightly increased Cmax of cefetamet. The Cmax values differed significantly (p<0.05) only between the cisapride group (5.76plus minus1.50 mg/L) and the N-acetylcysteine group (4.53plus minus1.18 mg/L). CONCLUSION: None of the small pharmacokinetic differences between the three groups is expected to have any relevance in the treatment of infectious diseases with cefetamet pivoxil. PMID- 11864101 TI - Colonization by vancomycin-resistant enterococci of the intestinal tract of patients in intensive care units from French general hospitals. AB - OBJECTIVE: To evaluate the prevalence of fecal carriage of vancomycin-resistant enterococci (VRE) by patients hospitalized in intensive care units from 24 French general hospitals. METHODS: Rectal swabs were obtained from 647 patients hospitalized in intensive care units during the month of June 1994 and plated on agar medium selective for vancomycin-resistant enterococci. The glycopeptide resistance phenotypes and genotypes of the enterococci detected were characterized. RESULTS: Thirty-two of 647 patients (4.9%) carried VRE. Thirteen strains (2%) were identified as Enterococcus faecium and 19 (2.9%) as Enterococcus gallinarum or Enterococcus casseliflavus. None of these strains was highly resistant to gentamicin. The E. gallinarum and E. casseliflavus strains contained the vanC1 and vanC2 genes, respectively. The E. faecium strains were highly resistant to vancomycin and teicoplanin and carried the vanA gene. No infection due to VRE was observed during the study period. Pulsed-field gel analysis of total DNA following digestion with SmaI or KspI from 13 VanA-type E. faecium strains revealed intra- and inter-hospital strain heterogeneity. However, the finding of isolates with indistinguishable pulsed-field types within the same ward and in two medical centers suggests patient-to-patient transmission or a common source. Four E. faecium strains were isolated within 48 h after admission of patients. CONCLUSIONS: These results indicate that VRE form part of the normal flora of patients and that, despite the actual scarcity of infections due to VRE, there is a potential risk for dissemination of these strains in French hospitals. PMID- 11864102 TI - Microbial etiology of acute community-acquired pneumonia in adult hospitalized patients in Yaounde-Cameroon. AB - OBJECTIVE: To determine the microbial etiology of acute community-acquired pneumonia in Yaounde. METHODS: Ninety-one consecutive adult patients admitted to hospital for radiologically confirmed acute community-acquired pneumonia were studied prospectively. Sputum microscopy and culture, blood cultures, pneumococcal antigen detection in serum and serologic analysis for agents of atypical pneumonia and for human immunodeficiency virus (HIV) were performed for most patients. RESULTS: There were 65 men and 26 women, mean age 36.5 years. Of 91 patients, 70.3% had at least one predisposing factor and 21.7% (20 of 81) were seropositive for HIV. A microbial etiology was identified in 48 (52.7%) cases. A single pathogen was identified in 42 (85.4%) and double pathogens in six (14.6%). Bacteremia occurred in 12 of 81 patients and was significantly more common in HIV seropositive than in HIV-seronegative patients. Streptococcus pneumoniae was the commonest causative agent, identified in 22 of 91 (24.2%) patients, 10 of whom were bacteremic. Atypical pathogens were diagnosed in 14 of 65 patients with serologic tests. Mycoplasma pneumoniae and Coxiella burnetii were diagnosed in six of 65 (9.2%) cases each, and Chlamydia pneumoniae in three (4.6%) patients. Mycoplasma pneumoniae and Chlamydia pneumoniae occurred as a dual infection in one case. Seven of 91 patients died, and death was not associated with any particular etiology. CONCLUSION: Streptococcus pneumoniae remains the predominant etiologic pathogen of community-acquired pneumonia. For this reason, and also because ampicillin used empirically to treat patients with this disease in the same setting has been shown to be efficacious, we propose the use of an aminopenicillin in the initial treatment of acute community-acquired pneumonia in adults in Yaounde. However, patients who fail to respond clinically to such treatment should benefit from either a macrolide or a tetracycline in order to cover for atypical pathogens. PMID- 11864103 TI - Chlamydia pneumoniae: a new opportunistic infectious agent in AIDS? AB - OBJECTIVE: To determine the incidence of Chlamydia pneumoniae respiratory tract infection in HIV-positive or AIDS patients. METHODS: Serum samples from 82 HIV positive patients with fever and respiratory symptoms were evaluated using microimmunofluorescence assay to detect C. pneumoniae-specific IgG and IgM antibodies. RESULTS: Twenty patients were found to have IgG antibodies to C. pneumoniae at titers ranging between 1:16 and 1:1024. Seven of the patients had evidence of acute C. pneumoniae infection (a fourfold rise in the titer of IgG antibody, or a single IgG titer of greater-than-or-equal1:512, or a single IgM titer greater-than-or-equal1:16). Five were diagnosed as having pneumonia and two bronchitis. No co-infection with other respiratory tract pathogens was found. CONCLUSIONS: Results of this study indicate that C. pneumoniae may play a role in the etiology of respiratory tract infections in HIV-positive patients; this fact should affect empirical antibiotic prescription. PMID- 11864104 TI - Invasive candidosis in pediatric patients. AB - OBJECTIVE: To analyze invasive Candida infections in pediatric patients and to examine the outcome of disease. METHODS: An observational prospective study was carried out of microbiologically documented cases of invasive candidosis in neonates, infants and children at Hippokration Hospital from December 1993 to July 1995. RESULTS: Thirty-nine cases of invasive candidosis (mainly candidemia and candiduria) occurred in 38 patients aged 3 days to 14 years, 21 (54%) having occurred in patients aged <1 month. The incidence was 10.1/1000 admissions in the two neonatal departments versus 1.1/1000 for all other pediatric departments, including oncology (p<0.01). The most frequent causes of hospitalization were premature birth or perinatal problems and surgery. Thirty-five strains of Candida albicans, 10 of C. parapsilosis and eight of other or unidentified species were isolated. Non-C. albicans isolates, especially C. parapsilosis and C. glabrata, exhibited higher minimal inhibitory concentrations of azoles as compared with C. albicans. Thirteen patients (34%), all candidemic, died within 0--40 days. Untreated patients more frequently had positive cultures up to time of death (p<0.0001) and died (p=0.0006). CONCLUSIONS: Invasive candidosis is frequent in pediatric patients, especially premature neonates, and is associated with increased mortality. With early diagnosis and initiation of therapy the outcome is favorable. PMID- 11864105 TI - Two clones of methicillin-resistant Staphylococcus aureus in Poland. AB - OBJECTIVE: To evaluate relatedness among methicillin-resistant Staphylococcus aureus (MRSA) strains isolated in Poland. METHODS: Ninety-three MRSA hospital isolates were collected from different regions in Poland from 1990 to 1992. Strains were analyzed with respect to heterogeneity of methicillin resistance, phage types, resistance patterns, crystal violet staining, chromosomal DNA SmaI restriction patterns by PFGE, ERIC1 and ERIC2 AP-PCR types and DNA repeat polymorphism within the protein A gene. Resistance to methicillin was confirmed by the detection of the mecA gene by PCR. RESULTS: The combined results of typing methods demonstrate that all MRSA strains analyzed could be easily divided into two distinct clones (clonally related strains). The first consisted of strains with clear heterogeneous expression of resistance to methicillin (34 isolates) and the second showed more homogeneous resistance (59 isolates). In this study the best method for epidemiologic analysis of MRSA was found to be PFGE. A good correlation between the epidemic behavior of MRSA and a high number of repetitive DNA units within the protein A gene was observed. CONCLUSIONS: Results show that in Poland two distinct clones of epidemic MRSA have circulated in the past, easily discriminated by pheno- and genotyping methods, and both could be found together in a single hospital. PMID- 11864106 TI - High frequency of antibiotic resistance among Gram-negative isolates in intensive care units at 10 Swedish hospitals. AB - OBJECTIVE: To investigate the resistance rates among Gram-negative isolates in Swedish intensive care units (ICUs). METHODS: During 1994-95, members of the Swedish Study Group collected, on clinical indication, 502 consecutive initial isolates of Gram-negative bacteria from patients admitted to ICUs at 10 Swedish hospitals and performed minimal inhibitory concentration (MIC) determinations with the Etest. Breakpoints were defined according to the criteria of the Swedish Reference Group for Antibiotics (SRGA). RESULTS: The distribution of bacterial species was: Escherichia coli > Klebsiella spp. > Enterobacter spp. > Pseudomonas aeruginosa > Haemophilus spp. > Proteus spp. > Stenotrophomonas maltophilia > Citrobacter spp. > Acinetobacter > Pseudomonas spp. > Morganella morganii > Serratia spp. Together these constituted 97% of all isolates. The frequencies of resistance for all the initial Gram-negative isolates were: ceftazidime 6.8%, cefotaxime 14.9%, ceftriaxone 18.5%, cefuroxime 44.1%, ciprofloxacin 4.2%, co trimoxazole 17.8%, gentamicin 5.8%, imipenem 8.6%, piperacillin 20.2%, piperacillin/tazobactam 12.9% and tobramycin 5.8%. CONCLUSIONS: Among Gram negative isolates in Swedish ICUs, a very high frequency of resistance was seen to cefuroxime, and rather high frequencies of resistance to cefotaxime, ceftriaxone, piperacillin and piperacillin/tazobactam. These drugs cannot be recommended for further use as empirical monotherapy for severe ICU-acquired Gram negative infections in ICUs in Sweden. PMID- 11864107 TI - Molecular epidemiology of Staphylococcus aureus strains colonizing the lungs of related and unrelated cystic fibrosis patients. AB - BACKGROUND: Cystic fibrosis (CF) patients can become persistently colonized with Staphylococcus aureus. This is initiated at an early age and may continue until or sometimes even during adolescence. Little is known about the epidemiology and cross-infectivity of S. aureus in CF patients, whether via the environment or person to person. METHODS: S. aureus isolates (n=189) from six unrelated CF patients and six pairs of CF siblings were genetically typed by arbitrary primed polymerase chain reaction (AP-PCR) assays. RESULTS: This longitudinal study revealed 35 different genotypes among the 189 isolates; the median number of types in a patient was three (range 1--6). One common S. aureus genotype was found in six patients and involved 20% of all isolates analyzed. Ultimately, in most of the patients long-term colonization with a single genotype was observed. In several, but certainly not all, pairs the siblings became persistently colonized with isolates that could not be discriminated by the typing method used; different S. aureus genotypes were isolated on an incidental but relatively frequent basis. Only one pair of siblings never shared identical isolates at any time during the screening period. CONCLUSIONS: In five of six cases, identical isolates were shared by CF siblings at a certain time. This suggests intra-family transmission or the presence of a common environmental source. The fact that in most of the CF sibling pairs different genotypes of S. aureus caused the ultimate long-term colonization indicates that, despite regular cross-colonization, patient characteristics select the S. aureus strain best adapted to the affected lung. Some genotypes may be particularly prevalent in the CF patient population, but additional studies are needed to confirm this. PMID- 11864108 TI - Computerized restriction endonuclease analysis compared with O-serotype and phage type in the epidemiologic fingerprinting of Pseudomonas aeruginosa strains. AB - OBJECTIVE: To assess restriction endonuclease analysis (REA) of chromosomal DNA using SalI enzyme, low-concentration (0.4%) agarose gels and digitalized data management of the REA patterns obtained for the typing of clinical Pseudomonas aeruginosa isolates. METHODS: A group of 67 clinical unrelated isolates from 10 Spanish hospitals was used to study the discriminatory power, reproducibility and typeability of REA typing. RESULTS: A SalI REA pattern consisted of a variety (1- 10) of restriction bands in the range between 12.2 and 48.5 kb and an unresolvable smear of low-molecular-weight bands. Forty different SalI REA patterns with an index of discrimination of 0.979 were obtained. Low typeability (91.04%) was the major limitation of REA typing. Analysis of blinded subcultures of eight Pseudomonas aeruginosa strains showed the reproducibility of REA typing to be 87.5%. Combined phenotypic typing (O-serotyping and phage typing) performed on the same group of strains showed comparable discrimination but much lower reproducibility. Isolates selected from five clusters of nosocomial infections in hospitals in the UK were typed by REA typing, and the results show high agreement when compared with conventional phenotypic typing methods in distinguishing between strains. CONCLUSIONS: These data underline the usefulness of REA typing enhanced with digitalized data management for the epidemiologic subtyping of clinical Pseudomonas aeruginosa isolates. PMID- 11864109 TI - Fingerprinting of Salmonella enterica subsp. enterica serovar Enteritidis by ribotyping. AB - OBJECTIVE: To carry out an epidemiologic evaluation of Salmonella enterica subsp. enterica serovar Enteritidis outbreaks in households and small communities by means of rRNA gene restriction pattern analysis (ribotyping). METHODS: One hundred Enteritidis isolates dating from 1989 to 1994 which could be allocated epidemiologically to different sources or to small community outbreaks were investigated with ribotyping, a fingerprinting method in which bacterial DNA is hybridized with the biotin-labeled plasmid pKK 3535 containing a ribosomal RNA operon of Escherichia coli to determine the ribosomal RNA gene restriction patterns. RESULTS: Four different ribotyping patterns were found with the restriction endonuclease Smal and nine with Sphl. Ribotypes of isolates which could be allocated epidemiologically to a common source usually corresponded. Almost 60% of the Enteritidis infections had the ribotyping pattern Sphl-A. In contrast, this pattern was not found in any of the five Enteritidis strains isolated in 1989. The suspicion that Enteritidis phage type 4 infections are caused by consumption of insufficiently heated eggs is supported by the fact that the ribotyping pattern Sph1-A was found in isolates from eggs and from human specimens. CONCLUSIONS: As patterns Sphl-A and Smal-J appeared in 58% and 75% of the isolates, respectively, ribotyping cannot be used for the differentiation between various outbreaks with these two patterns. In cases where the Enteritidis strains showed less frequent patterns, ribotyping seems to be a practical tool for the identification of infection chains. In addition newly appearing ribotyping patterns can give information about the epidemiologic development of Enteritidis infection. PMID- 11864110 TI - Serodiagnosis of tuberculosis and leprosy by enzyme immunoassay. AB - OBJECTIVE: To evaluate the use of serodiagnosis for tuberculosis and leprosy using mycobacterial antigen 38 kDa, with kits from Omega laboratories, to detect IgG by enzyme immunoassay (EIA). METHODS: The study population consisted of 58 patients with evidence of tuberculous infection (culture of Mycobacterium tuberculosis complex or microscopic evidence), of whom 23 had pulmonary and 35 had extrapulmonary disease. There were six subjects who had recently been treated for tuberculosis, 11 patients on treatment for leprosy and 137 patients suspected of having tuberculosis on clinical or radiologic grounds (without laboratory evidence). A control group comprised 35 healthy individuals or patients suffering from diseases other than tuberculosis. RESULTS: The tests showed that there was a significant difference in antibody levels between the patients with active pulmonary disease, extrapulmonary tuberculosis and leprosy in comparison with the control group (p<0.001). The sensitivities of the two tests together for proven pulmonary tuberculosis were 100% and 95.7% at 1.0--1.5 and >1.6 EIA cut-off points respectively, while the specificities were 88.5% and 100% at the same cut off points. The sensitivities for extrapulmonary tuberculosis were 71.4% and only 51.4% at 1.0--1.5 and >1.6 EIA cut-off points. The test was positive in 30 (21.9%) of the 137 suspected patients, while 43 (31.4%) had an equivocal result and the remaining 64 (47.7%) suspects were definitely negative. There was again a significant difference in positivity rates between suspects and the control group. CONCLUSIONS: Omega IgG test is useful in the serodiagnosis of active pulmonary tuberculosis and leprosy, but less sensitive in extrapulmonary disease, particularly in children. Equivocal results may only add to the evidence of tuberculosis in early or minimal disease. PMID- 11864111 TI - Detection of Bartonella henselae and Bartonella quintana by a simple and rapid procedure using broad-range PCR amplification and direct single-strand sequencing of part of the 16S rRNA gene. AB - OBJECTIVE: To detect directly Bartonella henselae and Bartonella quintana using culture-independent, molecular techniques, and to evaluate a simple and rapid procedure that allows uncultivable bacteria to be detected in usually sterile clinical specimens in a diagnostic laboratory. METHODS: From four clinical specimens proven to contain B. henselae (n=3) or B. quintana (n=1) DNA, part of the 16S rRNA gene was amplified using the polymerase chain reaction (PCR) and broad-range bacterial primers followed by reamplification and direct, single strand sequencing. The partial 16S rRNA sequences were compared to reference sequences in databases. RESULTS: Similarities between sequences derived from clinical samples and those of B. henselae and B. quintana, respectively, were in the range 98.7--100%, indicating a strong association to the genus Bartonella. Intraspecies sequence variations within the B. henselae sequences were observed. CONCLUSIONS: The method described is a rapid, sensitive and reliable tool to generate partial 16S rRNA sequences of B. henselae and B. quintana directly from normally sterile clinical specimens. It is compatible with adequate prevention of contamination as is needed for diagnostic purposes, and it possesses the potential to detect other pathogens, including those as yet unknown. PMID- 11864112 TI - Comparison of the Etest and a microbroth dilution system (Sceptor) to a reference agar dilution method for susceptibility testing of Bilophila wadsworthia. AB - OBJECTIVE: To compare the Etest and a microbroth dilution system (Sceptor) to a reference agar dilution method for susceptibility testing of Bilophila wadsworthia. METHODS: The susceptibility of 15 clinical isolates of Bilophila wadsworthia was determined by the National Committee for Clinical Laboratory Standards (NCCLS) agar dilution method using triphenyltetrazolium chloride for endpoint determination. The results were compared with the results obtained by the E test and a commercial microbroth dilution system (Sceptor). RESULTS: Comparison of the MICs obtained by the reference method and the Etest revealed few discrepancies, with piperacillin and metronidazole being the only exceptions. The overall agreement was 70% within one dilution step. The discrepancies did not result in major interpretative errors. The overall essential agreement using susceptibility categories was 98% for the E test and 99% for the microdilution system. CONCLUSIONS: Both methods may be considered as acceptable alternatives for testing individual isolates of B. wadsworthia. PMID- 11864113 TI - Serious Streptococcus pyogenes disease. PMID- 11864114 TI - Penicillium janthinellum in sputum and bronchoalveolar lavage in an AIDS patient with pneumonia. PMID- 11864115 TI - Enterococcal joint prosthesis infection. PMID- 11864116 TI - Treatment of disseminated Mycobacterium simiae infection in AIDS. PMID- 11864117 TI - Lyme disease presenting as prolonged pyrexia of unknown origin. PMID- 11864118 TI - Disseminated mucormycosis due to Absidia corymbifera in a patient with inflammatory bowel disease. PMID- 11864119 TI - Subinhibitory concentrations of gentamicin reduce production of listeriolysin, the main virulence factor of Listeria monocytogenes. PMID- 11864120 TI - Circumventing antibiotic resistance in specialized hospital units. PMID- 11864121 TI - Empirical antibacterial treatment for sepsis and the role of glycopeptides: recommendations from a European panel. PMID- 11864122 TI - Do the new lipid formulations of amphotericin B really work? AB - Over the past two years, there has been a dramatic rise in the prescription of lipid formulations of amphotericin B at our hospital. These compounds now account for a significant proportion of all expenditure on antimicrobial agents. We therefore conducted a review of the efficacy of the lipid formulations of amphotericin B. Only one randomized controlled trial has assessed the efficacy of any of these formulations in treating proven fungal infections, and this is only available in abstract form. Most of the available evidence on the use of lipid formulations is in the form of case series. There are therefore limited data to justify the widespread use of these compounds, and we believe that there are few circumstances when their administration is warranted. We suggest that local policies should be drawn up for the prescription of lipid formulations of amphotericin B, and, until more compelling data are available, that these drugs only be administered after discussion with microbiologists or infectious diseases physicians. PMID- 11864123 TI - Wide geographic distribution of a unique methicillin-resistant Staphylococcus aureus clone in Hungarian hospitals. AB - OBJECTIVE: To determine the genetic relatedness of methicillin-resistant Staphylococcus aureus (MRSA) isolates recovered from six provincial hospitals in Hungary between 1993 and 1994. METHODS: Molecular fingerprinting methods were used: hybridization with a mecA-specific DNA probe after ClaI restriction; hybridization with a probe for Tn554; and pulsed-field gel electrophoresis after Smal digestion of chromosomal DNA. RESULTS: All strains were resistant to penicillin, oxacillin, erythromycin, gentamicin, tetracycline, imipenem, and cephalosporins, and variably resistant to ofloxacin, clindamycin and tobramycin; all isolates were susceptible to vancomycin. Forty-eight of the 51 isolates carried the mecA gene as determined by Southern hybridization, using a mecA specific DNA probe, indicating that the methodology used for initial identification may have been in error in three of the cases. Forty-seven of the 48 mecA-positive isolates showed very similar genetic backgrounds as defined by pulsed-field gel electrophoresis (PFGE) patterns after Smal digestion of chromosomal DNAs: a unique PFGE pattern was seen in 32 isolates and minor variants of it in 15 additional isolates. All the 47 isolates carried the same mecA polymorph (Clal type III), as determined by DNA hybridization after Clal digestion of chromosomal DNA. Only one of the MRSA isolates had a completely different PFGE pattern and a novel mecA polymorph. CONCLUSIONS: The findings demonstrate the existence of a unique, epidemic MRSA clone, in both invasive and colonizing strains, which is widely dispersed in Hungarian hospitals hundreds of kilometers apart. PMID- 11864124 TI - Epidemiology of Staphylococcus aureus bacteremia in Denmark from 1957 to 1990. AB - OBJECTIVE: To investigate the changes in epidemiology of Staphylococcus aureus (SA) bacteremia in Denmark over a 30-year period, where the population has remained stable. METHODS: Bacteriologic and clinical data were generated on 17 712 SA strains from virtually all SA bacteremia cases in Denmark from 1957 to 1990 submitted to our laboratory for phage typing. The data were related to information about population, hospital activity and blood-culturing activity during that period. RESULTS: SA bacteremia cases increased from 3 to 20/100 000 inhabitants per year, with the largest increases in incidence rates for the <1 year and >50-year age groups. While blood-culturing activity increased three-fold during the period, the rate of SA bacteremias actually decreased relative to the number of blood cultures taken. The increase in SA bacteremia cases was mainly due to increases in nosocomial infections for all age groups and was related to the increasing admission rates to Danish hospitals. Major shifts in antibiotic resistance patterns and phage types took place during the period, i.e. a marked reduction in multiresistant (including methicillin-resistant) strains, but could not explain the change in the epidemiology of the infections. CONCLUSIONS: The data indicate that increases in SA bacteremia rates correlated significantly with increasing numbers of admissions to hospitals. The main increase in SA bacteremia rates was represented by nosocomial infection, although increasing blood culturing activity during the period may have contributed. PMID- 11864125 TI - Evaluation of tumor necrosis factor-alpha, interleukin-6 and C-reactive protein plasma levels as predictors of bacteremia in patients presenting signs of sepsis without shock. AB - OBJECTIVE: To evaluate the sensitivity of interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) plasma measurement to detect bacteremia in patients presenting sepsis signs, and to evaluate the potential benefit of such measurement in terms of early antimicrobial therapy initiation. METHODS: Plasma was obtained from 166 hospitalized patients for whom blood cultures were drawn for sepsis. Clinical data and antimicrobial therapies were noted. IL-6, TNF and C reactive protein (CRP) were measured. The sensitivities of these markers were retrospectively compared with the accuracy of the attending physician in initiating empirical antimicrobial therapy. The setting was an 850-bed university hospital. RESULTS: Thirty-four bacteremias and 69 non-bacteremic infections were noted. In 63 others, no infection was documented. Median (range) IL-6 plasma levels in the three groups of patients were 462 (15--50 850), 189 (<15--38 300) and 91 (<10--13 750) pg/mL, respectively (p<0.01). The corresponding TNF-alpha plasma levels were 37.5 (<15--2400), 15 (<15--240) and 15 (<15--200) pg/mL, respectively (p<0.01). CRP plasma levels were 10.7 (<0.6--30.2), 10.3 (<0.6- 34.4) and 7.3 (<0.6--20.9) mg/dL, respectively (p=0.12). With respect to these three parameters, IL-6 and TNF-alpha appear better than CRP for predicting bacteremia. Clinical features resulted in starting empirical antimicrobial therapy in only 62% of the bacteremic patients. On the other hand, 68% of these bacteremic patients had high IL-6 plasma levels (>200 pg/mL). A combination of clinical features and high IL-6 levels would have permitted early treatment for 82% of the bacteremic patients. CONCLUSIONS: IL-6 and TNF-alpha thus appear to be useful and earlier markers of bacteremia in septic patients. By contrast, CRP is neither sensitive nor specific in this setting. PMID- 11864126 TI - Ciprofloxacin treatment in chronic prostatitis. PMID- 11864127 TI - Molecular analysis of ampicillin-resistant sporadic Salmonella typhi and Salmonella paratyphi B clinical isolates. AB - OBJECTIVE: To study the mechanisms of antibiotic resistance in Salmonella typhi and Salmonella paratyphi B clinical isolates, and the clonality of resistant strains. METHODS: Antibiotic susceptibility was tested by disk-agar diffusion. Conjugation experiments and plasmid analysis by agarose gel electrophoresis after EcoRI digestion were followed by hybridization to a digoxigenin-labeled TEM-type beta-lactamase probe. DNA fingerprints were obtained by pulsed-field gel electrophoresis of Xbal-digested chromosomal DNA. RESULTS: Three S. typhi isolates (7% of the isolates studied), of which one was ampicillin resistant and the other two multiresistant (ampicillin, chloramphenicol, tetracycline, sulfamethoxazole/trimethoprim and streptomycin), and two ampicillin-resistant S. paratyphi B isolates (25% of the isolates studied) were further evaluated. A 34 MDa conjugative plasmid, previously isolated from Salmonella enteritidis, conferred ampicillin resistance. A 100-MDa conjugative plasmid encoded resistance to chloramphenicol, tetracycline and sulfamethoxazole/trimethoprim, as well as ampicillin. Chromosomal fingerprinting revealed two distinct resistant strains for each serovar which were different from a matched set of sensitive S. typhi strains. CONCLUSIONS: Two conjugative, TEM-type beta-lactamase-encoding plasmids conferred ampicillin resistance to S. typhi and S. paratyphi B. The 34-MDa plasmid was identical to that previously characterized from S. enteritidis, while the 100-MDa plasmid also encoded resistance to chloramphenicol, tetracycline and sulfamethoxazole/trimethoprim. Resistant isolates did not belong to a single clone but rather represented distinct strains. PMID- 11864128 TI - Carriage of group B Streptococcus in pregnant women and newborns: a 2-year study at Perugia General Hospital. AB - OBJECTIVE: To study the prevalence of group B Streptococcus (GBS) colonization in pregnant women and their newborns at Perugia General Hospital. METHODS: The number of mother---child pairs examined was 2300. Vaginal swabs were collected from the mothers at delivery, and auricular and pharyngeal swabs and gastric aspirate from the newborns at birth. Maternal risk factors for GBS disease, including premature delivery, intrapartum fever, prolonged rupture of membranes and multiple births, were evaluated. RESULTS: Maternal and neonatal colonization rates were 11.3% and 4.6%, respectively. GBS was isolated in 41.5% of the neonates born to colonized mothers and in 0.1% of those born to non-colonized mothers. No significant difference was observed in vertical transmission rates in the presence or absence of maternal risk factors. The external auditory canal was the most frequent (93.5%) and heavily colonized body site. Type Ib was the most common serotype among GBS isolates from mothers and babies. C surface protein was not detected in serotype V and VIII isolates, but was frequent in all other serotypes. Early-onset disease was observed in 0.4/1000 live births. CONCLUSIONS: The prevalence of maternal and neonatal colonization at Perugia General Hospital was similar to that obtained in other studies performed in Italy. The external auditory canal was confirmed as the most reliable body site to be sampled for the detection of neonates exposed to maternal GBS colonization. PMID- 11864129 TI - Ear infections with Shewanella alga: a bacteriologic, clinical and epidemiologic study of 67 cases. AB - OBJECTIVE: To present bacteriologic and clinical data on 67 patients from the island of Funen, Denmark, with Shewanella alga, a bacterium rarely seen in Scandinavia, isolated from ear swabs. Included in the study is an examination of the occurrence of S. alga in sea water around the island. METHODS: Bacteriologic examination and antibiotic susceptibility testing of 67 clinical isolates, 11 sea water isolates and two reference strains were conducted. Clinical information was obtained from the referring physicians. RESULTS: During 6 months S. alga was isolated from 67 patients, in 33 cases in pure culture. Seventy per cent of the patients were children between 3 and 15 years old who had clinical symptoms of acute or chronic otitis media. Previous ear disease was common (76%). Most of the cases (85%) occurred in August or September, and 47 of 55 patients reported contact with sea water shortly before symptoms developed. From seven of the patients, S. alga was isolated more than once. The species was also isolated from five of 10 bathing areas around the island of Funen. CONCLUSIONS: The patients were probably infected with S. alga during sea-water bathing in the unusually warm summer of 1994. Infections with marine bacteria are possible in countries with a temperate climate; patients with previous ear disease are at special risk. PMID- 11864130 TI - CQ-397 and CQ-414: antimicrobial activity and spectrum of two fluoroquinolone-- cephalosporin, dual-action compounds with carboxamido bonds. AB - OBJECTIVE: To evaluate the potential spectrum of activity of two novel dual action compounds with carboxamido bonds (CQ-397 and CQ-414; Laboratorios Aranda, San Rafael, Mexico) against human pathogens. METHODS: Approximately 800 Gram positive and Gram-negative aerobic clinical bacteria were tested in vitro using the Mueller-Hinton broth microdilution method of the National Committee of Clinical Laboratory Standards. RESULTS: CQ-397 (cefamandole+enrofloxacin) and CQ 414 (cefamandole+norfloxacin) were equally potent against Enterobacteriaceae (MIC90 range, 0.06--0.5 microg/mL and 0.06--1 microg/mL, respectively). Citrobacter freundii (MIC90, 4 microg/mL) and Providencia spp. (MIC90, >32 microg/mL) exhibited elevated study drug MICs. Enterobacteriaceae resistant to fluoroquinolones generally remained resistant. CQ-397 and CQ-414 were active against Stenotrophomonas maltophilia (MIC90, 4 microg/mL) and oxacillin susceptible staphylococci (MIC90, 0.25 microg/mL), but not oxacillin-resistant Staphylococcus aureus (MIC90, >32 microg/mL), Staphylococcus epidermidis (MIC90, 8 microg/mL), and enterococci (MIC90s, 8 to >32 microg/mL). There was no difference in the dual-action drug activity (MIC90, 2 microg/mL) between penicillin-susceptible and -resistant pneumococci. Haemophilus influenzae and Moraxella catarrhalis were very susceptible (MIC range, less-than-or-equal0.015- 0.06 microg/mL) to both compounds. CONCLUSIONS: The activity of these novel dual action compounds, formed from the bonding of older antimicrobials, warrants further investigation for potential human and/or animal health use, including toxicology and pharmacokinetics. PMID- 11864131 TI - Comparison of BacT/Alert and BACTEC NR 860 blood culture systems in a laboratory not continuously staffed. AB - OBJECTIVE: To evaluate the performance of continuously working blood culture systems in a discontinuous laboratory system. METHODS: The systems used were BacT/Alert (Organon Teknika Corp., Durham, NC) and BACTEC NR 860 (Becton Dickinson Diagnostic Instruments, Sparks, Md) in a comparison in a laboratory staffed 8 1/2 h on Mondays to Fridays and 4 1/2 h on Saturdays. Blood culture bottles (BacT/Alert aerobic and anaerobic, BACTEC NR 26 A and NR 27 A) were received thrice daily. RESULTS: From 1824 pairs of blood culture vials, 110 clinically significant microorganisms were recovered by both BACTEC and BacT/Alert, 43 by BACTEC alone, and 33 by BacT/Alert alone. The differences between the systems in total recovery and in recovery of individual species were not statistically significant. The average detection times were 13.36 h for BACTEC and 13.93 h for BacT/Alert (P>0.1). These times represent only 35.6% (BACTEC) and 32.6% (BacT/Alert) of the total timespans from collection of blood to informing the ward of a positive result (tcrd, clinically relevant detection time). If 24 h per day blood culture processing conditions and continuous transport of vials to the laboratory had been available, these percentages would have risen to 87% (BACTEC) and 87.5% (BacT/Alert). Under such 'ideal' conditions, ttrd could have been reduced by 22.16 h using BACTEC and by 26.81 h using BacT/Alert. The BacT/Alert system showed more false-positive results than the BACTEC system (80 (4.39%) versus 23 (1.26%), P<0.001). CONCLUSIONS: No time benefit for detection of positive blood cultures is gained with continuously measuring systems, if loading and processing of vials is organized discontinuously, as in our laboratory. PMID- 11864132 TI - Rapid recovery of Mycobacterium tuberculosis complex from clinical specimens using the BACTEC 9000 MB system, a new automated fluorimetric technique. AB - OBJECTIVE: To evaluate the new non-radioactive automated method BACTEC 9000 MB system for the rapid detection of mycobacteria in clinical specimens. METHODS: Ninety clinical specimens from 90 patients with a clinical diagnosis of tuberculosis were tested by both BACTEC 9000 and standard microbiological methods, and the results compared. RESULTS: The BACTEC 9000, in comparison with the standard method, showed significantly higher detection rates (45 of 90 positive versus 34), shorter time to culture positivity (mean time 18.8 versus 27.4 days) and lower contamination rate (2.2% versus 5.5%). CONCLUSIONS: These results encourage the use of this new system and suggest its use in microbiological laboratories involved in mycobacteriology. PMID- 11864133 TI - PCR-based DNA fingerprinting of Staphylococcus haemolyticus to investigate nosocomial infections. AB - OBJECTIVE: To apply PCR-based DNA fingerprinting in a clinical microbiology laboratory to investigate nosocomial infections with Staphylococcus haemolyticus. METHODS: DNA fingerprints were generated by PCR on 99 S. haemolyticus isolates using different primer combinations based on ERIC, REP or arbitrarily chosen simple repeat sequences. RESULTS: Primer combinations REP1+(GTC)6 and ERIC1+ERIC2 had sufficient discrimatory power and were chosen to analyze the clinical isolates. DNA fingerprint patterns from strains isolated from the patients nursed in the same hospital ward in the period 1991--94 were approximately 90% similar to each other. One staff member, sampled in 1991, carried a strain with a similar fingerprint. CONCLUSIONS: PCR based DNA fingerprinting is a suitable method to perform in a clinical laboratory. An S. haemolyticus strain appeared to be endemic in the hospital ward and had most probably been transmitted from patient to patient. S. haemolyticus may carry glycopeptide resistance and needs attention as a causative agent of nosocomial infections. PMID- 11864134 TI - Susceptibility testing by E-test and agar dilution of 30 strains of Legionella spp. isolated in Portugal. AB - OBJECTIVE: To evaluate the activity of erythromycin, roxithromycin, ciprofloxacin, doxycycline, sulfamethoxazole trimethoprim and rifampin against 32 strains of Legionella spp. under different testing conditions. METHODS: Minimum inhibitory concentrations (MICs) were determined by the E-test (Ab Biodisk, Solna, Sweden) and agar dilution reference technique (National Committee for Clinical Laboratory Standards (NCCLS), 1990) on two different media, buffered charcoal yeast extract agar (BCYE-alpha) and buffered yeast extract agar (BYE alpha), under 48- and 72-h incubation, without CO2. RESULTS: All the antimicrobial agents were inhibited by BCYE-alpha agar. The MIC90 values on BYE alpha were lower than those on BCYE-alpha but the variation factor was not the same: ciprofloxacin and rifampin, followed by erythromycin, suffered the greatest inhibition by the charcoal in the culture medium. Except for ciprofloxacin and rifampin, the 72-h MIC90 readings were always higher than the 48-h results whenever the agar dilution method was used. The E-test results showed slight variations with some, but not all, antibiotics. The most active agents against the 32 Legionella strains tested were rifampin and ciprofloxacin. CONCLUSIONS: BCYE-alpha is not suitable for susceptibility testing of Legionella spp. The E test method on BYE-alpha agar with 48-h incubation is recommended. PMID- 11864135 TI - Comparison of Auxacolor with API 20 C Aux in yeast identification. AB - OBJECTIVE: To compare Auxacolor with API 20 C Aux for identification of yeasts. METHODS: A total of 206 isolates belonging to 25 species was used in this study. Conventional yeast identification methods were used as a reference. RESULTS: With API 20 C Aux, the correct identification rate was 89.3% after 2 days, while 94.7% of the strains were correctly identified after 3 days. One of 14 strains of Candida tropicalis and 10 of 16 strains of Trichosporon cutaneum were not correctly identified. With Auxacolor, the percentages of correct identification after 1 and 2 days were 60.1% and 69.4%, respectively. Most strains of 11 of the 20 species considered in the system were correctly identified, including several of the most frequent yeast species. Several less commonly encountered yeast species were not correctly identified. Suggestions for improvement of the Auxacolor system are given. CONCLUSIONS: For the most frequent yeast species, Auxacolor, after adaptation and correction of the identification table, provides a useful alternative to API 20 C Aux. For less frequently encountered yeast species, the use of API 20 C Aux is preferable. PMID- 11864136 TI - Illustrated continuing medical education --- Case 1. PMID- 11864137 TI - Pharmacokinetics of 18F-labeled trovafloxacin in normal and Escherichia coli infected rats and rabbits studied with positron emission tomography. PMID- 11864138 TI - Intravitreous penetration of fluconazole during endophthalmitis. PMID- 11864139 TI - Pulsed-field gel electrophoresis (PFGE) pattern of genomic DNA of penicillin resistant pneumococci in Korea, where the resistance rate is very high. PMID- 11864140 TI - Resistance of Streptococcus pneumoniae to penicillin, erythromycin and third generation cephalosporins in Seville, Southern Spain. PMID- 11864141 TI - High-level aminoglycoside resistance and glycopeptide resistance among enterococci isolated from blood cultures, 1990--95. PMID- 11864142 TI - Long-standing bacteremia and endocarditis caused by Staphylococcus lugdunensis in a patient with an implantable cardioverter defibrillator. PMID- 11864143 TI - Clostridium difficile toxin A detection on colonies. PMID- 11864144 TI - Clostridium difficile toxin A detection on colonies. PMID- 11864145 TI - 1996 Report of the Comite de l'Antibiogramme de la Societe Francaise de Microbiologie. PMID- 11864146 TI - Review of meeting: European Study Group on Enterohemorrhagic Escherichia coli. PMID- 11864147 TI - Interactions between cytomegalovirus and the p53 tumor suppressor gene in atherosclerotic vascular disease. PMID- 11864148 TI - Broad- and narrow-spectrum antibiotics: an unhelpful categorization. PMID- 11864149 TI - Herpes esophagitis: a comprehensive review. PMID- 11864150 TI - Resistance to second- and third-generation cephalosporins among Escherichia coli and Klebsiella species is rare in Finland. AB - OBJECTIVE: To investigate the antimicrobial resistance of Escherichia coli and Klebsiella spp. from pus, urine and respiratory specimens, with particular emphasis on the detection of third-generation cephalosporin resistance. METHODS: E. coli (698) and Klebsiella sp. (476) strains from pus, respiratory and urinary specimens from hospital patients were collected from 19 laboratories. Data about consumption of third-generation cephalosporins and cefuroxime were collected from 24 hospitals. Antimicrobial susceptibility was tested with disk diffusion in primary laboratories and by an agar dilution method. Extended-spectrum beta lactamase (ESBL) production was studied with a double disk synergy test and an ESBL Etest. The beta-lactamase classes were characterized with polymerase chain reaction probes of the TEM and SHV beta-lactamase families and isoelectric focusing. RESULTS: Only 0.6% of E. coli and 2.3% of Klebsiella spp. strains were resistant or intermediately resistant to cefotaxime, ceftriaxone and/or ceftazidime. The ESBL producers detected comprised one E. coli harboring TEM-like genes and five Klebsiella pneumoniae strains, two of which harbored SHV-like genes, two TEM-like genes and one both. Although consumption of cefuroxime has increased in the years 1990-1994, from 3.48 to 5.84 defined daily doses/100 bed days, and the consumption of third-generation cephalosporins from 1.25 to 1.94 defined daily doses/100 bed-days, cefuroxime resistance of E. coli was only 3%. CONCLUSION: Although the use of broad-spectrum cephalosporins has increased, resistance to second- and third-generation cephalosporins is still rare in Finland. PMID- 11864151 TI - Increasing incidence and widespread dissemination of methicillin-resistant Staphylococcus aureus (MRSA) in hospitals in central Europe, with special reference to German hospitals. AB - OBJECTIVE: to present data on prevalence and interregional spread of methicillin resistant Staphylococcus aureus (MRSA) in Germany. METHODS: A nationwide collection of MRSA isolates from nosocomial infections in 143 hospitals was established from isolates (n=4368) sent to a microbiological reference center during 1993--95. As chosen by distinguishable resistance phenotypes at each time of occurrence during the study period, 1830 isolates were subjected to molecular typing by means of Smal macrorestriction patterns, PCR for RNA gene spacer patterns, and PCR for patterns of DNA stretches flanked by the ERIC-2 sequence and flanked by Tn916 and ribosomal binding site. In addition, data from a multicenter study on the incidence of antibiotic resistance have been analyzed (32 centers, 637 S. aureus isolates). RESULTS: In 1995 the prevalence of MRSA among S. aureus isolates was 8.7% overall in central Europe (including Germany), in comparison to 1.7% in 1990. From 1993 until now, a continuous interregional dissemination of six epidemic strains, which were identified by molecular typing, was recorded. Besides these epidemic strains, 15 MRSA strains were identified which could not be allocated to the epidemic MRSA or to the known clonal groups of the species S. aureus. MRSA from three cases of sporadic nosocomial infections exhibited characteristics of the clonal group of S. aureus with the capacity for toxic shock syndrome formation. The pattern of one MRSA corresponded to those of the S. aureus group exhibiting phage pattern 94,96. CONCLUSIONS: The prevalence of MRSA has increased in central Europe (and Germany) during the last 5 years, to 8.7%. The main source of infection with MRSA is obviously interregional dissemination of epidemic strains. At the same time, the mecA gene has been acquired by strains previously sensitive to methicillin. PMID- 11864152 TI - Association of distinct species of Borrelia burgdorferi sensu lato with neuroborreliosis in Switzerland. AB - OBJECTIVE: To evaluate by species-specific immunoblots the association of Borrelia burgdorferi sensu stricto, B. garinii and B. afzelii with neuroborreliosis in Switzerland. METHODS: Borrelia strains isolated from the cerebrospinal fluid (CSF) of three children with neuroborreliosis were typed by phenotypic and genotypic analysis. The serologic reactions (IgG) of these three patients as well as those of 28 patients, including one of these three children, with confirmed neuroborreliosis were characterized and scored by immunoblots on the three individual Borrelia species antigens. Twenty patients with typical erythema migrans served as a control group. RESULTS: Phenotypic and genotypic analysis confirmed that all three CSF isolates were B. garinii. In the 28 patients with neuroborreliosis, the comparatively strongest reactions were as follows: 18 to B. garinii, three to B. burgdorferi sensu stricto and two to B. afzelii; five were inconclusive. In the control group (erythema migrans), the comparatively strongest reactions were as follows: six B. garinii, one to B. burgdorferi sensu stricto and five to B. afzelii; eight were indeterminate. CONCLUSIONS: Typing of these three CSF isolates and characterization by immunoblots of the antibody reactions of patients with neuroborreliosis give additional evidence of the association of B. garinii and neuroborreliosis. Our serologic results suggest that B. burgdorferi sensu stricto and B. afzelii are also responsible for some neuroborreliosis cases in Switzerland. Our immunoblots and the scoring system proved particularly useful for the serologic typing of patients with late Lyme borreliosis. PMID- 11864153 TI - Influence of macrolides on mucoid alginate biosynthetic enzyme from Pseudomonas aeruginosa. AB - OBJECTIVE: The long-term administration of erythromycin (EM), clarithromycin (CAM) or azithromycin (AZM) has generally resulted in a favorable outcome for patients with diffuse panbronchiolitis (DPB) infected with mucoid Pseudomonas aeruginosa. To elucidate the mechanism involved, the influence of macrolides on mucoid alginate production by P. aeruginosa was investigated in vitro. METHODS: The macrolides used in this study were EM with a 14-membered ring, AZM with a 15 membered ring, midecamycin (MDM) with a 16-membered ring, and CP-4305, which has had mycarose removed from MDM, The effects of macrolides on mucoid P. aeruginosa were investigated by quantitative assay of alginate production and inhibition of guanosine diphospho-D-mannose dehydrogenase activity. RESULTS: After incubation with EM, AZM and CP-4305, the structural material of P. aeruginosa biofilm was distorted, and the enzymatic activity of GDP-D-mannose dehydrogenase, the most important enzyme in mucoid alginate biosynthesis, was inhibited. However, these effects were not observed with the 16-membered macrolide MDM. CONCLUSIONS: The basic mechanism of clinical efficacy seen characteristically in 14- or 15 membered macrolides for patients with airway biofilm disease depends on the ability of such macrolides to inhibit alginate production by P. aeruginosa. Furthermore, this suggests that the inhibitory effect observed with 14-, 15- and 16-membered macrolides may depend on the sugar chain connected with the macrolide ring. PMID- 11864154 TI - Efficacy and safety of vancomycin constant-rate infusion in the treatment of chronic Gram-positive bone and joint infections. AB - OBJECTIVE: To evaluate the efficacy and safety of vancomycin constant-rate infusion over 24 h in the treatment of Gram-positive bone infections, METHODS: Vancomycin (40 mg/kg/day) was administered without a loading dose to 15 patients (12 male, three female) aged 23--90 years, weighing 46--85 kg, with postoperative chronic bone and joint infections. The 24-h dose was adjusted to maintain plasma levels between 25 and 35 mg/L. Mean duration of therapy was 6.2 months (4--8.5) via a portable infusion pump. Sites of infection included hip and femur (10), tibia (three), patella (one) and vertebrae (one). Sequestrectomy (two), removal of material (7/8 prosthetic hips, 1/5 metal implants) and debridement (two) were performed at the beginning of the treatment. Involved bacteria included Staphylococcus aureus (eight, six methicillin resistant), S. epidermidis (four methicillin-resistant), Enterococcus faecalis (one), Enterococcus avium (one) and Streptococcus bovis (one). RESULTS: MIC of vancomycin ranged from 1 to 4 mg/L. The mean vancomycin bone concentration when available was 67.7plus minus38.9 microg/L. Based on a mean post-treatment follow-up of 14plus minus4 months (6- 20.6), cure was achieved in 10 patients (66.6%). Failures were related to the inability to remove the infected prosthesis (one) or implants (three) and to the persistence of a deep wound abcess (one). Adverse events included pruritus (four cases), tinnitus (two), mild transient elevation of creatinine level (three) and transient neutropenia (two). Vancomycin was maintained in all the patients. CONCLUSIONS: Prolonged treatment with vancomycin constant-rate infusion is effective and safe for treatment of Gram-positive chronic bone and joint infections, providing that complete surgical debridement and prosthetic material removal are performed. PMID- 11864155 TI - Efficacy of 5-day cefpodoxime proxetil for recurrent pharyngitis in adults. A comparative study with 10-day penicillin V or amoxycillin---clavulanate. AB - OBJECTIVE: To compare the clinical and bacteriologic efficacy of a 5-day course of cefpodoxime proxetil (CPD) with that of a 10-day course of penicillin V (PNV) or amoxycillin---clavulanate (AMC) in recurrent pharyngitis in adults. A cost effectiveness study (reported elsewhere) was carried out at the same time. METHODS: This multicenter, randomized, open label trial involved 580 adult patients consulting general practitioners for clinical recurrent pharyngitis (greater-than-or-equal3 episodes within the last 12 months) regardless of the bacterial etiology. Patients were treated for 5 days with CPD, 100 mg twice daily, or for 10 days with PNV, 1x106 IU three times a day, or for 10 days with AMC, 500 mg (amoxycillin) three times a day. Clinical and bacteriologic outcomes were noted at the end of treatment, and cases of clinical recurrence were recorded during a 6-month follow-up period. RESULTS: At the end of treatment, clinical response was satisfactory in 157 of 170 (92.3%) patients on CPD, 147 of 166 (88.5%) patients on PNV, and 168 of 177 (94.9%) patients on AMC. Group A beta hemolytic streptococci (GABHS) were eradicated in 22 of 23 (95.65%) patients on CPD, 16 of 16 (100%) patients on PNV, and 19 of 20 (95%) patients on AMC. The rates of clinical success and GABHS eradication were not significantly different between the groups. Compliance (p<0.001) and tolerance (p<0.001) were significantly better in the CPD group than in the other two groups. Among the 389 patients evaluable 6 months after the end of treatment, the recurrence rate of acute pharyngitis (due to any bacterium) was significantly lower in the CPD group (p=0.01 versus PNV; p<0.01 versus AMC). A Kaplan---Meier analysis (469 patients over 6 months) of the rate of non-recurrence, with comparison by the log-rank test, also showed a significant difference in favor of CPD. CONCLUSIONS: A 5-day treatment of recurrent pharyngitis with CPD was as effective and better tolerated than a 10-day treatment with PNV or AMC. The risk of recurrence was lower with CPD. PMID- 11864156 TI - Toxicity and efficacy of conventional amphotericin B deoxycholate versus escalating doses of amphotericin B deoxycholate---fat emulsion in HIV-infected patients with oral candidosis. AB - BACKGROUND: Amphotericin B deoxycholate remains the treatment of choice for most systemic fungal infections; however, its clinical use can be limited by infusion related side effects and nephrotoxicity. New formulations of amphotericin in lipid compounds have been shown to decrease toxicity. We previously showed that a lipid emulsion preparation of amphotericin B deoxycholate was better tolerated than the conventional preparation in dextrose. Therefore, we have now studied the clinical tolerance, renal toxicity and efficacy of higher doses of amphotericin B deoxycholate prepared and infused in a fat emulsion (Intralipid 20%). Thus, this report adds information to the previous publication. METHODS: Forty-two patients infected with HIV and suffering oral candidosis entered the study. The patients received either amphotericin B deoxycholate---glucose 1 mg/kg/day or amphotericin B deoxycholate---lipid emulsion 1 mg/kg/day for 4 days (randomized phase), or amphotericin B deoxycholate---lipid emulsion 2 mg/kg/day or 3 mg/kg/day (escalating-dose phase) for 5 days. Clinical (immediate) side effects and renal (creatinine) tolerance were assessed daily; efficacy against oral candidosis was measured by using a simple clinical score. Serum levels of amphotericin B were also measured. RESULTS: None of the patients receiving amphotericin B deoxycholate---lipid emulsion had treatment interrupted, as compared to four (36%) in the amphotericin B deoxycholate---glucose group (pless-than-or equal0.01); chills during or after the infusions were significantly less frequent in the amphotericin B deoxycholate---lipid emulsion groups than in the amphotericin B deoxycholate-glucose group (p=0.03). The increase of creatininemia during treatment was significantly higher for patients receiving amphotericin B deoxycholate---glucose than for those receiving amphotericin B deoxycholate-- lipid emulsion (p=0.001). The number of patients who had a creatininemia greater than-or-equal18 mg/L during treatment was significantly higher in both the amphotericin B deoxycholate---glucose group (36%) and in the group receiving the highest dose of amphotericin B deoxycholate---lipid emulsion than in other groups (pless-than-or-equal0.06). The serum concentrations of amphotericin B were lower for the amphotericin B deoxycholate---lipid emulsion regimen than for the amphotericin B deoxycholate---glucose regimen at the same dose of 1 mg/kg/day, but increased with the dose. The change of the oral candidosis score was similar for the same dose of 1 mg/kg/day of amphotericin B deoxycholate infused in either glucose or lipid emulsion; higher doses of amphotericin B deoxycholate---lipid emulsion were more efficacious (p=0.009) and this efficacy seemed to increase with the dose (p=0.06). CONCLUSIONS: The clinical and renal tolerance of amphotericin B deoxycholate are improved when the drug is directly prepared and infused in lipid emulsion (Intrapid) and this preparation allows for greater dosage, up to 3 mg/kg/day, with resultant greater efficacy. This preparation is simple and cost-effective (approximately 7 US $ per 50 mg of amphotercin B) and could be clinically compared to other formulations of amphotericin B. PMID- 11864157 TI - Polymerase chain reaction (PCR) compared with conventional identification in culture for detection of Bordetella pertussis in 7153 children. AB - OBJECTIVE: To compare the polymerase chain reaction (PCR) results with conventional culture results for the diagnosis of Bordetella pertussis infections. METHODS: PCR and culture were performed in the course of a large vaccine efficacy trial in Germany on specimens taken from 7153 children less-than or-equal2 years of age with cough illness lasting >6 days, and laboratory results were compared with clinical data also obtained from the patients. Calcium alginate nasopharyngeal swabs were taken for culture and clinical data were obtained from patients. Swabs were inoculated on charcoal horse blood agar plates containing cephalexin, and then discarded. The agar plates were preincubated for 2 days at the physician's office and then shipped to the culture laboratory at the University Children's Hospital in Munich, Germany, for diagnosis of B. pertussis and B. parapertussis infections. In this laboratory, Dacron swabs for PCR were taken from each culture by a wide sweep over the culture. Swabs for PCR were stored in NaCl and sent weekly to the PCR laboratory at the University Children's Hospital in Basel, Switzerland, for PCR diagnosis of B. pertussis infections. RESULTS: B. pertussis was identified by culture in 3% (213/7153) and by PCR in 7.6% (546/7153) of the specimens. Therefore, PCR increased the identification rate of subjects with B. pertussis infection 2.6-fold. Clinical characteristics were considered according to the type of laboratory findings: group 1 consisted of 209 culture-positive and PCR-positive subjects, and group 2 of 337 culture-negative but PCR-positive subjects. Group 2 subjects were significantly more likely to have mild or atypical clinical symptoms of whooping cough than were group 1 subjects. By analyzing the PCR results of group 2 subjects semiquantitatively, it could be shown that the degree of PCR positivity correlated with the severity of the clinical symptoms of whooping cough in the patient. CONCLUSIONS: PCR identified many pertussis cases with mild or atypical clinical symptoms that were not identified by culture. Semiquantification of PCR products revealed that the less positive the PCR result, the higher was the failure rate in diagnosing pertussis by culture, and, in addition, the less typical were the clinical symptoms in the patient. PMID- 11864158 TI - Multicenter evaluation of mycobacteria growth indicator tube (MGIT) compared with the BACTEC radiometric method, BBL biphasic growth medium and Lowenstein---Jensen medium. AB - OBJECTIVE: To evaluate the new BBL mycobacteria growth indicator tube (MGIT) in comparison with other media. METHODS: MGIT was evaluated in 10 Italian centers on 433 clinical samples, mainly of respiratory origin and mainly smear positive, in comparison with Lowenstein---Jensen and with one or more other methods represented, according to participating centers, by the BACTEC radiometric method or by the biphasic BBL Septi-Chek AFB system. While MGIT and Lowenstein---Jensen were used for all the samples, 285 of them were also inoculated in BACTEC vials and 274 in biphasic bottles. Of these samples, 132 were investigated with all the four methods. RESULTS: Although less rapid and sensitive than the radiometric method, the results of MGIT were equal when compared with the other two media with respect to overall isolation yield; furthermore, it allowed the detection of growth in significantly shorter times. CONCLUSIONS: The results of this study indicate the value of MGIT for the detection of mycobacteria and, thanks to its extreme simplicity of use, its suitability for small and large laboratories. Its combined use with a solid medium can substantially improve the diagnosis of mycobacterial infection. PMID- 11864159 TI - Evaluation of the E-test for routine testing of the susceptibility of Streptococcus pneumoniae to benzylpenicillin, amoxicillin and cefotaxime. AB - OBJECTIVE: To study the routine use of the E-test for susceptibility testing of penicillin-resistant Streptococcus pneumoniae. METHODS: A multicenter study of penicillin-resistant S. pneumoniae (PRSP) was carried out in Brittany, France (10 general hospitals, and two university hospitals including a coordinating center). Each hospital detected PRSP by the oxacillin (5- micro g) disk method and determined the MICs of penicillin G, amoxicillin and cefotaxime by the E-test under routine conditions. All the PRSP strains were collected in a coordinating center and the MICs were checked by the agar dilution method. The classifications obtained from the MICs determined by the E-test and by the reference method were compared. RESULTS: Between 1 July 1993 and 30 June 1994, 128 PRSP strains were collected. Agreement within 1 log2 dilution was obtained for only 62% of strains with benzylpenicillin, 72.5% with amoxicillin and 76% with cefotaxime. These data are well below published values. In addition, 52% of the strains found to be penicillin-resistant by the reference technique were of intermediate resistance according to the E-test. There were major differences in the quality of the results obtained by the participating laboratories. CONCLUSIONS: There are problems of standardization in the routine use of the E-test. Microbiologists should therefore take particular care when performing the test and when reading the results, and ensure that reference strains are included in the assay. PMID- 11864161 TI - Listeria and listeriosis. PMID- 11864160 TI - Rapid detection of Clostridium difficile toxin A in stool specimens. AB - OBJECTIVE: To evaluate a rapid (15-min) enzyme immunoassay in the format of an individual cassette (ImmunoCard toxin A, Meridian, BMD, Marne-la-Vallee, France) for the detection of Clostridium difficile toxin A in stool specimens. METHODS: We compared this new test with the cytotoxicity assay using MRC-5 cells, the ToxA test (TechLab, BioWhittaker, Fontenay-sous-bois, France) and toxigenic culture for the diagnosis of C. difficile-associated diseases (CDAD). A total of 236 stool specimens collected from 220 patients was simultaneously tested with the four methods. Discordant results were resolved by reviewing patients' clinical records. RESULTS: The prevalence of CDAD was 13.9%. Test sensitivities and specificities were 100% and 99% respectively for the cytotoxicity assay, 87.5% and 100% for ImmunoCard toxin A, 77.4% and 100% for the ToxA test and 100% and 98% for toxigenic culture. CONCLUSIONS: The ImmunoCard Toxin A is a very rapid, individual and easy-to-perform test for the diagnosis of CDAD. It provides same day results and may be useful for both guiding appropriate treatment and controlling nosocomial spread of C. difficile. PMID- 11864162 TI - Illustrated continuing medical education. PMID- 11864163 TI - Bacteremia in children: a 2-year review of experience with a pediatric blood culture protocol using BacT/Alert standard bottles. PMID- 11864164 TI - Activity of eight fluoroquinolones against enterococci. PMID- 11864165 TI - Usefulness of delayed hypersensitivity skin tests in HIV infected patients. PMID- 11864166 TI - HIV RNA load and opportunistic infections. PMID- 11864167 TI - Cytomegalovirus seminal shedding in healthy volunteer sperm donors. PMID- 11864168 TI - In vitro activity of antibiotics and combinations against Brucella melitensis. PMID- 11864169 TI - Imported trichinellosis from former Yugoslavia. PMID- 11864170 TI - Group B streptococcus: rapid intrapartum detection and influence of density of maternal colonization on vertical transmission. PMID- 11864171 TI - Trichosporon beigelii peritonitis in a child on continuous ambulatory peritoneal dialysis. PMID- 11864173 TI - Viral and atypical pathogens as causes of type 1 acute exacerbations of chronic bronchitis. PMID- 11864172 TI - The challenge of infectious diseases: the European perspective. PMID- 11864174 TI - The clinical microbiologist's contribution to audit, quality and guidelines. PMID- 11864175 TI - An audit of microbiology laboratory utilization: the diagnosis of infection in orthopedic surgery. AB - OBJECTIVE: To analyse the quality of ordering, collection and transport of specimens for microbiological analysis by a department of orthopedic surgery. METHODS: The analysis consisted of a prospective formal evaluation performed by two consultant microbiologists. RESULTS: One hundred and seventeen consecutive requests were audited. These requests belonged to 55 clinical episodes, 39 of which were of (presumed) infection and 16 of surveillance. The main sites sampled were: joint 28 (51%), and extra-articular bone or tissue 6 (11%). Of 98 surgical specimens, 20 (20%) yielded a relevant microorganism. The requests were classified as definitely appropriate in 67% and 85% of episodes, by the two consultants respectively. No request was considered unjustified. Collection, handling and transport were categorized as definitely appropriate in 56% and 73% of requests. Analysis of compliance with an existing protocol for prosthetic joint revision revealed similar errors. CONCLUSION: Audits of this type can give invaluable information about the area of uncertainty between the clinician and the laboratory and can identify appropriate measures for corrective action. PMID- 11864176 TI - Suppurative mediastinitis after open-heart surgery: a comparison between cases caused by Gram-negative rods and by Gram-positive cocci. AB - OBJECTIVE: To compare clinical characteristics and risk factors of suppurative postsurgical mediastinitis according to its etiology. METHODS: Suppurative postsurgical mediastinitis developed in 45 (2.5%) of 1779 patients who underwent open-heart surgery at the Hospital Virgen del Rocio in Seville, Spain, from 1986 to 1996. Microbiological diagnosis was available in 42 patients. RESULTS: Gram negative rods were isolated in 19 cases and Gram-positive cocci in 23 cases. Seventeen isolates (38%) were sensitive to the antimicrobial agent used perioperatively. Patients with Gram-negative rod infection had a longer duration of bypass (127plus minus36 min versus 96plus minus34 min, p<0.01), and a worse postoperative condition. Longer mechanical ventilation (4plus minus7 days versus 1plus minus2 days, p<0.05) and concomitant infection in a remote site (pulmonary and/or urinary infection) were more frequently observed in this group than in patients with Gram-positive infections (58% versus 22%, p<0.05). Twenty patients (51%) were bacteremic. The mortality rate was 20% (five of 45). CONCLUSIONS: Preventable postoperative remote-site infection may lead to mediastinitis, especially if Gram-negative rods are involved. PMID- 11864177 TI - Isolation and virulence factors of verocytotoxin-producing Escherichia coli in human stool samples. AB - OBJECTIVE: To evaluate the isolation rate of O157 and non-O157 verocytotoxin producing Escherichia coli (VTEC) strains, to study the occurrence of additional virulence factors and to correlate these with clinical symptoms. METHODS: Over more than 5 years, 17 296 unduplicated fecal samples submitted for routine culture were screened for VTEC by a single PCR detecting VT1, VT2 and its variants. Verocytotoxin B subunit genotypes of the isolates obtained by testing individual colonies in positive samples were determined by a polymerase chain reaction---restriction fragment length polymorphism (PCR---RFLP) technique, the eaeA gene and the 60-MDa virulence plasmid by PCR, and the hemolytic phenotype by using CaCl2-washed blood agar. RESULTS: Verocytotoxin genes were found in 1.02% of the samples. Non-O157 VTEC strains were isolated in 0.66% and O157 in 0.17%. Overall, VTEC was less frequently isolated than Campylobacter and Salmonella but more frequently than Yersinia and Shigella. All cases except two siblings were epidemiologically unrelated. Cases of hemolytic uremic syndrome (HUS) were only observed in association with serogroup O157, which seems to be more pathogenic than the non-O157 strains. Among non-O157 VTEC strains, eaeA-positive strains are more frequently associated with clinical symptoms than are eaeA-negative strains. Other virulence factors correlate less closely with the presence of symptoms. CONCLUSIONS: VTEC is the third bacterial intestinal pathogen in our study population. All stool samples from patients with diarrhea should be screened for the most frequent serogroup, O157, or, if this is not possible, at least those from patients with bloody diarrhea. Non-O157 VTEC strains, especially if they are eaeA positive, are also associated with diarrhea, more often non-bloody. PCR or the new commercially available immunoassays could be used in selected cases, e.g. in patients suffering from HUS and in cases of outbreaks. PMID- 11864178 TI - Typhoid fever due to a Salmonella typhi strain of reduced susceptibility to fluoroquinolones. AB - OBJECTIVE: To report a case of typhoid fever contracted in Portugal in 1994 due to a Salmonella typhi isolate which had reduced susceptibility to fluoroquinolone (MIC 1 mg/L of ciprofloxacin) and high level resistance to nalidixic acid (MIC greater-than-or-equal 56 mg/L). METHODS: Molecular studies of reduced susceptibility to fluoroquinolones comprised complementation tests with a wild type allele and sequencing directly from PCR products of the gyrA gene. RESULTS: Complementation tests and DNA sequencing showed that a mutation occurred in the gyrA gene of this clinical isolate, resulting in a substitution of phenylalanine for serine at position 83 of GyrA. CONCLUSIONS: Because quinolones may be regarded as a treatment of choice in typhoid fever, it seems important now to recommend cautious use of these drugs as first-line therapy and possibly use of nalidixic acid resistance as a marker for detection of 'first-step' resistance to fluoroquinolones in S. typhi. PMID- 11864179 TI - Acquired drug resistance in Mycobacterium tuberculosis isolates from treated patients in the province of Bursa, Turkey. AB - OBJECTIVE: To evaluate the distribution of acquired resistance in isolates of Mycobacterium tuberculosis from treated patients in two periods, 1984--89 and 1990--95, in the Bursa (Southern Marmara) region. METHOD: Susceptibility of 531 M. tuberculosis isolates to four commonly used drugs (isoniazid (INH), streptomycin (SM), ethambutol (EMB) and rifampin (RMP) was determined by the absolute concentration method of Canetti et al. RESULTS: In 203 strains isolated in the years 1984--89, the total acquired resistance was 32.5%, and it was 37.5% in 328 strains isolated in 1990--95 (p>0.05). Resistance to INH, SM, RMP and EMB was found in 23.6%, 16.7%, 6.4% and 3.9%, respectively, in the first period (1984 -89), and in 26.2%, 20.4%, 25.3% and 8.2%, respectively, in the second period (1990--95). The increase in RMP resistance was statistically significant (p<0.001). The incidence of multidrug-resistant strains was 12.3% in the first period, and 24.4% in the second period, a significant increase (p<0.001). CONCLUSIONS: We believe that progressive emergence of phenotypes resistant to INH+RMP in our region is caused by inadequate treatment for various reasons. In the present study, the fact that multidrug resistance occurred in nearly 25% of patients treated previously but still infective suggests that the approach to surveillance, patient therapy and follow-up programs should be fundamentally reconsidered in our region. PMID- 11864180 TI - A simple and reliable method to screen isolates of Escherichia coli and Klebsiella pneumoniae for the production of TEM- and SHV-derived extended spectrum beta-lactamases. AB - OBJECTIVE: To evaluate which of 24 beta-lactams used in susceptibility tests best discriminated between strains of Klebsiella pneumoniae and Escherichia coli that produce extended spectrum beta-lactamases (ESBLs) from strains that produce older, more familiar, plasmid-mediated beta-lactamases such as TEM-1 and SHV-1. METHODS: Susceptibility to the 24 beta-lactam agents was determined by agar dilution and disk diffusion methodologies, using 27 strains of K. pneumoniae and E. coli that produced 22 different older plasmid-mediated beta-lactamases and 28 strains that produced 17 different ESBLs. RESULTS: In general, strains that produced ESBLs were intermediate or resistant to cefpodoxime, whereas those that produced other beta-lactamases were susceptible to this agent. The agar dilution test exhibited 96% sensitivity and 100% specificity in discriminating these two groups of organisms. The disk diffusion test exhibited 100% sensitivity and 96% specificity. All other beta-lactam agents tested were inferior discriminators between the two groups of organisms. CONCLUSIONS: Agar dilution and disk diffusion tests with cefpodoxime can be used to discriminate strains of K. pneumoniae and E. coli that produce ESBLs from those that produce older, plasmid mediated beta-lactamases. PMID- 11864181 TI - Efficacy of a selective and differential medium for isolating Corynebacterium urealyticum from urine specimens. AB - OBJECTIVE: To describe a new selective and differential medium for improving the isolation of Corynebacterium urealyticum from urine. METHODS: A total of 370 urine specimens, 270 from patients with underlying urologic disease and 100 from to a control group without renal pathology, were cultured on standard sheep blood agar medium and on a new selective medium for Corynebacterium urealyticum. RESULTS: The use of this selective medium resulted in a nine-fold increase in the prevalence of bacteriuria caused by C. urealyticum (12.4% versus 1.4%). Furthermore, the frequency of patients with symptomatic bacteriuria detected with the selective medium (3.2%) was higher than with the non-selective medium (1.4%). CONCLUSIONS: We recommend the use of this selective medium for the isolation of C. urealyticum in patients with underlying urologic disease or with chronic urinary symptoms. PMID- 11864182 TI - Comparison of agar-based media for primary isolation of glycopeptide-resistant enterococci. AB - OBJECTIVE: To compare four vancomycin-containing agar media for the isolation of glycopeptide-resistant enterococci (GRE) from clinical fecal specimens: kanamycin --aesculin---azide (KAA) agar; bile---aesculin---polymixin (BAP) agar; aztreonam- -amphotericin blood (CBAA) agar; and neomycin blood (CBN) agar. METHODS: Fecal specimens from 125 patients were inoculated onto each medium. Media were examined for enterococci after incubation for up to 48 h. Enterococci were identified to species level, and glycopeptide phenotypes were determined by measuring minimum inhibitory concentrations of vancomycin and teicoplanin. RESULTS: GRE were isolated from 44/125 samples. Enterococcus faecalis and Enterococcus faecium isolates, expressing glycopeptide resistance of the VanA or VanB phenotypes, were recovered from 27/33 (82%) specimens on BAP medium, 26/33 (79%) on KAA medium, and 21/33 (64%) on CBN and CBAA media. Enterococcus gallinarum and Enterococcus casseliflavus isolates expressing low-level glycopeptide resistance (VanC phenotype) were recovered from 14/15 (93%) specimens on CBAA medium, 7/15 (47%) on KAA and CBN media, and 6/15 (40%) on BAP medium. CONCLUSIONS: The media tested in this study, with the exception of CBN medium, detected at least 75% of patients colonized by GRE. Further development of BAP, CBAA and KAA media is warranted to improve growth and selectivity. PMID- 11864183 TI - Rabies. PMID- 11864184 TI - Illustrated continuing medical education. PMID- 11864185 TI - The human granulocytic ehrlichiosis (HGE) agent in Swedish ticks. PMID- 11864186 TI - Multiple intracranial tuberculomas in a non-immunocompromised patient. PMID- 11864187 TI - Reduced susceptibility to quinolones in Salmonella typhi acquired in Europe: a clinical failure of treatment. PMID- 11864188 TI - Endocarditis caused by Enterococcus faecalis with high-level resistance to aminoglycosides: failure of ampicillin and ceftriaxone combined therapy. PMID- 11864189 TI - Enterococcal endocarditis in a patient with a renal oncocytoma. PMID- 11864190 TI - Mycobacterium celatum as a cause of disseminated infection in an AIDS patient. PMID- 11864191 TI - Recovery of Mycobacterium haemophilum skin infection in an HIV-1-infected patient after the start of antiretroviral triple therapy. PMID- 11864192 TI - Molecular epidemiology of tuberculosis in a prison facility in Seville: a 3-year study (1993--95). PMID- 11864193 TI - Gingivocrevicular transudate for HIV screening. PMID- 11864194 TI - More on HBe Ag-negative mutants in chronic HBV infection. PMID- 11864196 TI - Report of the Comite de l'Antibiogramme. PMID- 11864195 TI - Controlling glycopeptide-resistant enterococci. PMID- 11864197 TI - Comparison of A60 and three glycolipid antigens in an ELISA test for tuberculosis. PMID- 11864198 TI - Response to Dr Maes. PMID- 11864199 TI - Atypical respiratory pathogens. AB - The main atypical pathogens in respiratory tract infections are classified on the basis of their ability to cause atypical pneumonia. This is not a well-defined clinical entity, and it is evident that atypical pathogens can sometimes cause 'typical' pneumonias and vice versa. This emphasizes the need for microbiological diagnosis, since it affects the selection of proper treatment, in which beta lactam antibiotics and aminoglycosides are not effective. Moreover, mixed infections caused by atypical and typical pathogens together are common. At this moment rapid and sensitive diagnostic methods are lacking. Besides numerous viruses, the main bacterial pathogens causing atypical pneumonias are Mycoplasma pneumoniae, two chlamydial species, Chlamydia pneumoniae and C. psittaci, one rickettsia, Coxiella burnetti, and several Legionella species. The majority of these pathogens cause upper respiratory tract infections more often than overt pneumonias. An atypical agent, Chlamydia pneumoniae, has also been associated with chronic inflammatory conditions in the cardiovascular system. The most recently discovered pathogen in atypical pneumonias is a hantavirus causing hantavirus pulmonary syndrome. PMID- 11864200 TI - The role of interleukin-10 in the pathogenesis of bacterial infection. PMID- 11864201 TI - In vivo efficacy of cefotaxime and amoxicillin against penicillin-susceptible, penicillin-resistant and penicillin---cephalosporin-resistant strains of Streptococcus pneumoniae in a mouse pneumonia model. AB - OBJECTIVE: To compare cefotaxime (CTX) to amoxicillin (AMO) (usually considered the definitive therapy for penicillin-susceptible Streptococcus pneumoniae infections) in an immunocompromised mouse pneumonia model. METHODS: Three S. pneumoniae clinical isolates were used: two serotype 19 strains, a penicillin susceptible (Ps) strain (penicillin MIC=0.03 microg/mL) and a highly penicillin resistant (Pr) strain (penicillin MIC=4 microg/mL), and one serotype 23F strain, a penicillin---cephalosporin-resistant (CFTR) strain (CTX MIC=4 microg/mL). RESULTS: CTX activity in this mouse model of pneumonia induced by the highly penicillin-resistant strain of S. pneumoniae was lower than expected from its low MIC against this organism. Furthermore, AMO had greater efficacy than CTX against a CFTR S. pneumoniae strain. CONCLUSION: Our data suggest that there is no major difference in the in vivo efficacy of the two agents, cefotaxime and amoxicillin, against penicillin-resistant and penicillin---cephalosporin-resistant S. pneumoniae. PMID- 11864202 TI - Salmonella bloodstream infection in Ghanaian children. AB - OBJECTIVE: To examine the frequency of community-acquired salmonella bloodstream infection in Ghanaian children and the occurrence of antibiotic resistance in salmonellae. METHODS: The study comprised 472 patients with a blood culture obtained within 48 h of admission to the pediatric department of Korle Bu Teaching Hospital in Accra, Ghana, over a 3-month period. All Salmonella isolates from blood cultures were speciated and antibiotic susceptibility tests were performed. Clinical data of children with salmonella bloodstream infection were compared to those of controls. Two control groups were identified: all children enrolled in the study without salmonella bloodstream infection (group 1), and those with bloodstream infection due to other organisms (group 2). RESULTS: A pathogen was isolated from 111 children (23.5%), and salmonellae were among the most common isolates (n=24; 21.6%). Among Salmonella strains, S. enteritidis (n=14; 59%) predominated over S. typhi (n=6; 25%). Resistance to several antibiotics was only found in S. enteritidis isolates (n=8; 57%). Children with salmonella bloodstream infection presented more often than controls with severe anemia, jaundice, abdominal pain and distension as well as hepatomegaly and splenomegaly. They were also hospitalized for a significantly longer period, but the case-fatality rate was similar. CONCLUSIONS: Salmonella bloodstream infection, especially due to non-typhoidal strains, is a potential health problem for Ghanaian children and may be complicated by resistance to the commonly available antibiotics. PMID- 11864203 TI - Multiresistant Pseudomonas aeruginosa serogroup O:11 outbreak in an intensive care unit. AB - OBJECTIVE: To determine whether 15 multiresistant Pseudomonas aeruginosa isolates from an intensive care unit (ICU) outbreak were related, were endemic, and belonged to the O:12 European clone. METHODS: Forty-six P. aeruginosa isolates from a large hospital were investigated with respect to their antibiotic resistance profiles, serogroups, bacteriocin types and DNA fingerprints obtained by pulsed-field gel electrophoresis (PFGE) of genomic DNA digested with Xbal. RESULTS: Fourteen of the ICU outbreak isolates were indeed identical with respect to their serogroup, O:11, pyocin type, 10/a, and PFGE type, A. Clone A was endemic and dominant throughout the hospital, even though, within the ICU, it underwent phenotypic alterations, such as loss of cell wall lipopolysaccharide side-chains, or acquisition of ceftazidime and imipenem resistance. Bacteriocin typing was more discriminatory than serotyping, but PFGE could differentiate further among phenotypically identical strains. It also allowed the tracking of an O:6 strain, as it was becoming gradually more resistant and undergoing a bacteriocin-type conversion while remaining genotypically unaltered. CONCLUSIONS: Using three typing methods, a nosocomial multiresistant strain distinct from the previously described dominant European O:12 clone was characterized, and the ability of PFGE to identify clonal isolates even when these appear phenotypically distinct was demonstrated. PMID- 11864204 TI - Low doses of zidovudine plus didanosine are less effective than higher doses of didanosine monotherapy: a randomized trial in patients pretreated with zidovudine. AB - OBJECTIVE: To compare the clinical efficacy and tolerance of didanosine (ddl) monotherapy with low-dose zidovudine/didanosine (AZT/ddl) therapy among HIV infected patients previously treated with AZT. METHODS: A randomized controlled trial was carried out of ddl 400 mg daily versus AZT/ddl 300/200 mg daily among patients with CD4 cell counts 0.05 in all cases). Subtype 1b was present in six of the seven cases of cirrhosis (85.7%) and in nine of the 18 cases of active chronic hepatitis (50.0%). PMID- 11864208 TI - Changes in reimbursement policy for antibiotics and prescribing patterns in general practice. AB - OBJECTIVE: To study the effect of a reduction in reimbursement of the cost of antibiotics on the prescribing pattern in primary care in Denmark. METHOD: We analyzed the general practitioners' prescriptions of antibiotics during 1993--96 in relation to a reduction in reimbursement on the basis of national health service data in the county of North Jutland (population 488 000). On 1 January 1996 the reimbursement for tetracyclines was withdrawn, and for other antibiotics reimbursement was reduced from 75% to 50%. RESULTS: The total consumption of all antibiotic groups increased steadily in the county until 1995, and in 1996 a decrease of 13% was seen. A very marked reduction was noticed immediately after 1 January 1996 for the more expensive broad-spectrum antibiotics. The use of tetracyclines dropped by 42% during the first 3 months of 1996 after withdrawal of reimbursement. CONCLUSIONS: It is reasonable to assume that the new reimbursement policy has initiated a reduction and caused a shift in general practitioners' prescribing of antibiotics. Thus a differential reimbursement policy might influence general practitioners' prescribing behavior towards antibiotics, with desirable ecological consequences. PMID- 11864209 TI - Detection of IgA and low-avidity IgG antibodies for the diagnosis of recent active toxoplasmosis. AB - OBJECTIVE: To determine the clinical value of testing IgA and the avidity of IgG (by two commercial systems) for the detection of recent active toxoplasmosis (RAT), and to study the IgG avidity during the course of infection. METHODS: The IgA was tested by a capture ELISA (Pasteur, France) and the avidity of IgG was determined by two modified commercial indirect ELISA methods (Sorin, Italy; Behringwerke, Germany) in 12 patients who were not immunosuppressed (group I) and 57 healthy subjects with a past infection by Toxoplasma gondii (group II). RESULTS: IgA was present in 75% of patients from group I and 21% of subjects from group II. The reliability for diagnosis of RAT was: sensitivity 75%, specificity 84%, positive predictive value 52.9% and negative predictive value 93.3%. In group I, 91.7% of patients had more than 50% low-avidity IgG, by both methods; in group II, 21% of subjects had low-avidity IgG at levels from 40% to 50%, by both methods. The diagnostic reliability of the two methods for the detection of low avidity IgG in the first samples of RAT was similar when a breakpoint of 50% was used, with values of: sensitivity 91.7%, specificity 100%, positive predictive value 100% and negative predictive value 98%. CONCLUSIONS: The study of IgA is not on its own adequate for diagnosis of RAT. However, testing the avidity of IgG is more reliable for the diagnosis of RAT, in studies of one serum sample or sequential samples. PMID- 11864210 TI - Multicenter comparative evaluation of two rapid immunoassay methods for the detection of Chlamydia trachomatis antigen in endocervical specimens. AB - OBJECTIVE: To evaluate two rapid immunoassay methods, QuickVue-Chlamydia (Quidel Corp., San Diego California) and Kodak SureCell (Kodak Corp., Rochester, NY) for the detection of Chlamydia trachomatis antigen in endocervical swabs from high- and low-risk females. METHODS: Seven hundred and twenty-four females attending three clinics were enrolled in the study. The results were compared to McCoy's or BGMK cell culture and discrepancies resolved with polymerase chain reaction and direct fluorescent antibody tests performed on left-over culture specimens. RESULTS: The sensitivity, specificity, predictive value of a positive and predictive value of a negative of the QuickVue Chlamydia assay were 92.0%, 99.1%, 92.0% and 99.1%, respectively. The sensitivity, specificity, predictive value of a positive and predictive value of a negative of the SureCell assay were 90.0%, 99.8%, 98.6% and 98.8%, respectively. CONCLUSIONS: The performances of the two immunoassay methods were similar, and slight differences in sensitivity and specificity were not statistically significant. Both immunoassay methods performed well in high- and low-risk patient groups, both for symptomatic and for asymptomatic patients. PMID- 11864212 TI - Evaluation of the Roche Amplicor polymerase chain reaction assay for detection of enteroviruses in cerebrospinal fluid and its potential impact on patient management. AB - OBJECTIVE: To evaluate the Roche Amplicor polymerase chain reaction assay (APCR) by comparing the detection of enteroviruses from cerebrospinal fluid (CSF) by the Roche assay with detection by viral culture and to determine whether routine use of enteroviral PCR will affect patient management. METHODS: One hundred and sixty three CSF specimens were tested by APCR and viral culture. Some of the discrepant specimens were resolved by retesting with an in-house PCR assay. Other discrepant results were resolved by testing the patients' serum by APCR or by viral culture of throat and stool specimens. RESULTS: Thirty CSF specimens were positive by APCR, and 18 of these were positive by viral culture. There were no APCR negative, viral-culture-positive CSF specimens. Six of the 12 discrepant specimens were resolved as true positives. CONCLUSIONS: The APCR assay was more rapid and sensitive than viral culture for detection of enteroviruses from CSF. Routine use of this assay has the potential to reduce the amount of antibiotics used and the number of patient days spent in hospital. PMID- 11864211 TI - Detection of Mycobacterium tuberculosis in mixed broth cultures using DNA probes. AB - OBJECTIVE: To investigate the ability of a commercial acridinium ester-labeled DNA probe (AccuProbe, Gen-Probe Inc., USA) to detect the presence of Mycobacterium tuberculosis complex in a mixed culture with Mycobacterium avium complex. METHODS: The density of organisms required to produce a positive result for Mycobacterium tuberculosis complex alone in broth culture was compared with the density required to produce a positive result in the presence of Mycobacterium avium complex. RESULTS: A threshold density of 1.5 x 106 CFU/mL was required for detection of Mycobacterium tuberculosis and this threshold remained unaltered in the presence of Mycobacterium avium complex. The presence of Mycobacterium avium complex had no effect on detection of Mycobacterium tuberculosis in a mixed broth culture incubated and probed over a 21-day period. CONCLUSIONS: The findings of the study suggest that the presence of Mycobacterium avium complex has no effect on the detection of Mycobacterium tuberculosis and that the Accuprobe test is potentially capable of detecting a dual infection with organisms of both complexes. PMID- 11864213 TI - Human infection with Bartonella species. PMID- 11864214 TI - A continuing problem of tetanus in the southeast of Turkey. PMID- 11864215 TI - Endocarditis in a Dutch patient caused by Bartonella quintana. PMID- 11864216 TI - Primary leptomeningeal lymphoma simulating chronic meningitis. PMID- 11864217 TI - Corynebacterium pseudotuberculosis infection in a butcher. PMID- 11864218 TI - Rhabdomyolysis and group B streptococcus meningitis in an elderly patient with hairy cell leukemia. PMID- 11864219 TI - Pasteurella multocida abcess in an African patient with human immunodeficiency virus infection. PMID- 11864220 TI - High levels of resistance to antibiotics in Escherichia coli isolated from Cambodian doctors during a postgraduate course. PMID- 11864221 TI - Phenotypes of resistance to macrolide and lincosamide antibiotics in Staphylococcus species. PMID- 11864223 TI - Acknowledgment of Reviewers for Volume 3. PMID- 11864222 TI - Nasopharyngeal carriage of Streptococcus pneumoniae in children's day-care centers: 10-month follow-up study in Nice, France. PMID- 11864225 TI - Unanswered questions concerning antibiotic resistance. PMID- 11864224 TI - Message From The Editorial Office. PMID- 11864226 TI - Serum levels of beta2-microglobulin, neopterin, TNF-alpha and soluble receptors of TNF-alpha and interleukin-2 in intravenous drug abusers according to HIV-1 status and stage of the HIV-1 infection. AB - OBJECTIVE: To compare the serum levels of beta2-microglobulin (beta2M), neopterin (NP), TNF-alpha and soluble receptors of TNF-alpha (sTNF-R55 and sTNF-R75) and interleukin-2 (sIL-2R) in a population of intravenous drug abusers according to whether or not they had HIV-1 infection and to the stage of the HIV-1 infection. METHODS: A cross-sectional study was performed at four drug detoxification centers in Barcelona, and the HIV outpatient clinic at Hospital Clinic in Barcelona. Three cohorts of intravenous drug abusers (IVDAs)-105 HIV-1-uninfected patients (cohort A), 174 with asymptomatic HIV-1 infection (cohort B) and 39 with AIDS (cohort C)-were enrolled. On the first visit, the following laboratory tests were performed: hemogram and platelet count, hepatitis B surface antigen (HBsAg), anti-hepatitis C antibodies, B2M, NP, sIL-2R, TNF-alpha, and TNF receptors (sTNF R55 and sTNF-R75). RESULTS: The three cohorts were homogeneous according to sex, type of drug, average number of intravenous doses of drug in 1 day, and hepatitis B infection. Patients with AIDS were older than those of cohort A and B (p<0.0001). HIV-negative IVDAs were co-infected with hepatitis C virus less frequently than were HIV-positive IVDAs (80% versus 91%, p<0.03). Among HIV-1 negative IVDAs (cohort A), almost all (from 86% to 95%, depending on the marker) individual values were within the normal boundaries of our laboratory. With a single exception (level of sTNF-R55 in cohort B compared with cohort A, p=0.15), levels of all markers were significantly higher in asymptomatic HIV-1-infected (cohort B) when compared with uninfected patients (cohort A), and in AIDS patients (cohort C) when compared with both cohorts A and B. There was a significant positive correlation between levels of ss2M and NP (r=0.56; p<0.01), ss2M and TNF (r=0.65, p<0.01) and NP and TNF (r=0.76, p<0.01). There was no correlation between levels of sIL-2R and levels of ss2M, NP or TNF and its receptors. CONCLUSIONS: Intravenous drug abuse does not modify serum levels of ss2M, NP, sIL-2R, TNF-alpha, and TNF receptors (sTNF-R55 and sTNF-R75). Levels of these markers increase significantly when an HIV-1 infection occurs and when there is progression to AIDS. PMID- 11864227 TI - Epidemiology of penicillin resistance in Streptococcus pneumoniae isolates in eastern France. AB - OBJECTIVE: To assess the rates of intermediate and high-level resistance to penicillin among Streptococcus pneumoniae isolates and to identify clonal relationship of isolates within the different serotypes by means of pulsed-field gel electrophoresis. METHODS: We studied all clinical isolates obtained between April 1995 and March 1996 from patients admitted to 10 hospitals in eastern France. Antibiotic susceptibility testing and serotyping were performed on all isolates. The genetic polymorphism of isolates susceptible, intermediately resistant and highly resistant to penicillin was studied by using pulsed-field gel electrophoresis with ApaI and SmaI endonucleases. RESULTS: The prevalence of intermediate and high-level resistance was respectively 30.3% and 9.7%. Diminished sensitivity to penicillin was mainly encountered in serotypes 6, 9V, 14 and 23F. The 9V isolates from the different hospitals were genetically closely related, unlike the 23F isolates. Different levels of resistance (MICs from 0.5 to 2 mg/L) were expressed by closely related isolates. Three 9V isolates, three capsular-type 14 isolates and one non-typeable isolate were genetically closely related in studies with the two endonucleases. CONCLUSIONS: The capsular type was not a good indicator of genetic relatedness. The level of penicillin resistance was independent of the clonal classification. Horizontal gene transfer may be the main factor determining the degree of resistance. PMID- 11864228 TI - Pharmacokinetics in vivo and pharmacodynamics ex vivo/in vitro of meropenem and cefpirome in the Yucatan micropig model: continuous infusion versus intermittent injection. AB - OBJECTIVE: To investigate the pharmacodynamic disposition of two recently developed beta-lactam antibiotics, meropenem and cefpirome, in the Yucatan micropig model, and to compare the bactericidal activity of these drugs against bacteria in this in vitro/ex vivo micropig model after administration by both intermittent injection and continuous infusion. METHODS: Cefpirome (1 g) was given to the micropig over a 12-h period by direct intravenous injection and 6-h continuous infusion (500 mg). Meropenem (250 mg) was administered either by 30 min intravenous and 8-h continuous infusion. The two drugs were assayed by HPLC. The pharmacodynamics of these drugs were evaluated by means of (1) serum killing curve against Klebsiella pneumoniae producing extended-spectrum beta-lactamase, stably derepressed Enterobacter cloacae and methicillin-susceptible penicillinase producing Staphylococcus aureus, and (2) calculations of index of surviving bacteria (ISB). RESULTS: The bactericidal activity of meropenem against K. pneumoniae and E. cloacae in this in vitro/ex vivo model was excellent, with a 4 log decrease at peak concentrations. Meropenem produced a mixed concentration- and time-dependent, killing effect against E. cloacae and K. pneumoniae. The ISB value ranged from 25% to 30% for E. cloacae. With concentrations above MIC for S. aureus (1 mg/L), cefpirome has a time-dependent bactericidal activity, as shown by the ISB ranging from 20% to 80% after 4 h and between 20% and 40% after an 8-h drug exposure. For both antibiotics, the higher concentrations obtained just after intermittent injection had a rapid and strong killing effect against the strains tested, but the trough levels had no bactericidal activity. The continuous infusions produce consistent concentrations of antibiotic that can be maintained above the MIC, and the bactericidal activity of which ranges from 2 to 4 log10 decrease of inoculum. CONCLUSIONS: In the present study the micropig has been shown to be an adequate model for the pharmacodynamic investigation of cefpirome and meropenem. In general, continuous infusion appears to optimize the pharmacodynamic profile of the two tested beta-lactam antibiotics. However, against Gram-negative bacilli, the administration of a loading dose prior to continuous infusion of beta-lactams would eliminate the only potential pharmacokinetic disadvantage of continuous infusion and ensure the rapid onset of antimicrobial activity. PMID- 11864229 TI - Characterization of coagulase-negative staphylococci by primer-specific polymerase chain reaction and ribotyping. AB - OBJECTIVE: To analyze, by primer-specific polymerase chain reaction (PCR) and ribotyping, coagulase-negative staphylococci (CNS). METHODS: Forty-five clinical isolates of CNS were identified by the API ID32 STAPH system and ribotyping. Additionally, primer-specific PCR was evaluated for identification of clinical strains of Staphylococcus epidermidis. RESULTS: Forty-five isolates of CNS from neonates with nosocomial bacteremia were studied. The results of the S. epidermidis-specific PCR were compared with those obtained using ribotyping and the API ID32 STAPH system. Excellent congruence was found between primer-specific PCR and ribotyping. Primer-specific PCR proved to be a fast and reliable method for the identification of S. epidermidis strains. According to the primer specific PCR and ribotyping analysis, a few CNS isolates were found to be incorrectly identified by the API ID32 STAPH system. CONCLUSIONS: Primer-specific PCR is a fast and reliable method for the identification of S. epidermidis. Primer-specific PCR in combination with ribotyping is a promising approach for studying the epidemiology of S. epidermidis and other CNS species in hospital. PMID- 11864230 TI - Assessment of the BacT/Alert blood culture system: rapid bacteremia diagnosis with loading throughout the 24 h. AB - OBJECTIVE: To determine blood culture (BC) diagnostic speed when combining an automated BC system with rapid loading of inoculated bottles throughout the 24 h. METHODS: A total of 111 positive BCs representing bacteremia were investigated in retrospect. All bottles were loaded into the BacT/Alert BC system (Organon Teknika) as soon as possible after sampling and time from specimen collection to Gram stain result was recorded. RESULTS: The mean time from specimen collection to loading was 3.5 h (median 2.1 h). We found that 74% of all positive BCs collected during daytime (08.00-16.00) were reported (as Gram stain) to the clinician before 17.00 the next day. For specimens collected between 16.00 and midnight the corresponding proportion was 67%. BCs drawn between midnight and 08.00 were reported before 17.00 the same day in 24% of the cases. CONCLUSIONS: Rapid loading of an automated BC system throughout the 24 h results in fast diagnosis of bacteremia. The diagnostic speed in this study represents a fair estimation of the maximal diagnostic speed accomplishable in a clinical situation with the BacT/Alert system in conjunction with normal daytime laboratory working hours. PMID- 11864231 TI - Enteric infections due to Escherichia coli. PMID- 11864233 TI - Clinical spectrum of infections due to Propionibacterium acnes. PMID- 11864232 TI - Problems in diagnosis and treatment of Staphylococcus haemolyticus endocarditis in a hemodialysis patient. PMID- 11864234 TI - Reinfection with Mycobacterium tuberculosis in an urban tuberculosis hospital. PMID- 11864235 TI - Catheter-related bacteremia due to Tsukamurella pulmonis. PMID- 11864236 TI - Prevalence of stably derepressed class I beta-lactamase in multiresistant clinical isolates of Enterobacter cloacae in two Hungarian hospitals. PMID- 11864237 TI - Broad- and narrow-spectrum antibiotics: a different approach. PMID- 11864239 TI - A SWAB in The Netherlands. PMID- 11864238 TI - The Society for Health Care Epidemiology of America. PMID- 11864240 TI - The Second International Symposium in Infection Models in Antimicrobial Chemotherapy. PMID- 11864241 TI - Mycobacterial infection models. PMID- 11864242 TI - Rabbit model of bacterial meningitis. PMID- 11864243 TI - Animal models of co-infection. PMID- 11864244 TI - High rates of antibiotic resistance among normal fecal flora Escherichia coli isolates in children from Greece. AB - OBJECTIVE: To study the distribution of antibiotic resistant Escherichia coli in the fecal flora of healthy children in Greece. METHODS: Rectal swabs were collected from 181 children, not suffering from infections and not undergoing antibiotic treatment, aged 6 months to 6 years, outpatients of a pediatric hospital, and plated on McConkey agar with ampicillin or trimethoprim. Isolated resistant colonies were identified to the species level and E. coli strains were studied further by molecular methods. RESULTS: Forty-four per cent of the children carried resistant E. coli, and in 20% resistance was transferable. Forty seven per cent of the children with no history of antibiotic consumption during the last year were found to carry resistant strains in their feces, and transferable R plasmids were present in 23% of them. Forty per cent of the strains and 30% of the transconjugants were multiresistant. Although plasmids of various molecular weights and restriction endonuclease digest patterns were identified, six 60-MDa and four 80-MDa plasmids, originating from epidemiologically unrelated children, were found to be similar. CONCLUSION: Normal flora E. coli in Greece seems to constitute an important reservoir of resistance genes. Eradication of resistance from a population that comes into frequent contact with antibiotics seems to be difficult. PMID- 11864245 TI - Bactericidal activity of three beta-lactams alone or in combination with a beta lactamase inhibitor and two aminoglycosides against Klebsiella pneumoniae harboring extended-spectrum beta-lactamases. AB - OBJECTIVE: To study the bactericidal activity of beta-lactam antibiotics (imipenem, cefepime, cefpirome) alone or in combination with a beta-lactamase inhibitor (sulbactam) in the presence or absence of aminoglycoside (amikacin or isepamicin) against Klebsiella pneumoniae strains producing extended-spectrum beta-lactamases (ESBLs). METHODS: We characterized 10 strains by means of analytic isoelectric focusing and pulsed-field gel electrophoresis. The ESBLs produced by these strains were derived from either TEM (TEM-1, TEM-2) or SHV-1. The killing-curve method was used for this bacterial investigation. Bacteria (final inoculum 5x105 CFU/mL) were incubated with antibiotics at clinical concentrations obtained in vivo. RESULTS: All the combinations with cefepime or cefpirome + sulbactam were bactericidal, with a 4 log10 decrease being obtained within 6 h without regrowth at 24 h, whereas imipenem alone, and combinations, gave a bactericidal effect within 6 h. The two cephalosporins alone decreased the inoculum of 4 log10 at 6 h but regrowth was observed at 24 h. When the aminoglycoside was added, this bactericidal effect was obtained within 3 h with amikacin and within 1 h with isepamicin. CONCLUSIONS: Cefepime + sulbactam or cefpirome + sulbactam may be an alternative to imipenem for the treatment of patients with ESBL-producing K. pneumoniae. Aminoglycosides are often associated in nosocomial infections due to ESBL-producing K. pneumoniae: isepamicin acted faster than amikacin, but both worked well. To conclude, it may be prudent to avoid extended-spectrum cephalosporins as single agent when treating serious infections due to ESBL-producing K. pneumoniae. Addition of a beta-lactamase inhibitor such as sulbactam +/- aminoglycoside is advisable to avoid failure of treatment. PMID- 11864246 TI - HIV-1-infected patients with focal neurologic signs: diagnostic role of PCR for Toxoplasma gondii, Epstein-Barr virus, and JC virus. AB - OBJECTIVE: To evaluate nested PCR for Toxoplasma gondii (TOX), JC virus (JCV) and Epstein-Barr virus (EBV) for diagnosis of toxoplasmic encephalitis (TE), progressive multifocal leukencephalopathy (PML) and primary central nervous system lymphoma (PCL). METHODS: A prospective study encompassed 26 HIV-1-infected individuals presenting with focal neurologic signs and symptoms. Nested PCR was performed on both supernatants and pellets of centrifuged cerebrospinal fluid (CSF), on plasma and on white blood cells (WBCs). For a retrospective study, stored CSF supernatants were available from an additional 27 HIV-1-infected patients with TE, PML, and PCL. RESULTS: TE, PML or PCL was diagnosed in 13 of 26 patients in the prospective group. Plasma and WBC analysis by PCR was not informative except in one case of TE. TOX and JCV were detected by PCR in the CSF pellets of four of five patients with TE, and of four of five patients with PML, respectively, but in no other cases. EBV was detected not only in three of three cases of PCL, but also in six patients suffering from other conditions. PCR on the CSF supernatants was less sensitive for all three etiologies. These results correlated with those of the retrospective PCR analysis, for which only stored CSF supernatants were available, revealing sensitivities of 33%, 50% and 66% for TE, PML and PCL, respectively, but specificities of 100%. CONCLUSIONS: In the clinical routine, TOX and JCV PCR on centrifuged CSF pellets can be recommended to obtain an early diagnosis of TE and PML. Under these conditions, EBV PCR helps to exclude PCL as a cause of FBLs, as it is highly sensitive, but not specific, for PCL in HIV-1-infected individuals. PMID- 11864247 TI - In vitro selective concentrations of cefepime and ceftazidime for AmpC beta lactamase hyperproducer Enterobacter cloacae variants. AB - METHODS: A mixed culture of a wild-type ceftazidime/cefepime-susceptible (4x107 CFU/mL) strain and an ampC derepressed Enterobacter cloacae (105 CFU/mL) strain (relative proportions 99.75% and 0.25%) was challenged for 4 h with different antibiotic concentrations of ceftazidime and cefepime (0.03-4096 mg/L), and then transferred to drug-free medium. The proportion of wild-type versus derepressed population was evaluated after 24 h. RESULTS: Ceftazidime and cefepime selected the derepressed variant at concentrations ranging from 1 to 4096 and from 0.12 to 16 mg/L respectively. CONCLUSIONS: These results suggest that serum concentrations attainable with a 2 g/8 h cefepime dosage may be able to suppress the emergence of derepressed ampC mutants. PMID- 11864248 TI - Efficacy of subunit trivalent influenza vaccine in previously vaccinated children suffering from hemophilia. AB - OBJECTIVE: To assess humoral immune response to subunit trivalent influenza vaccine in children suffering from hemophilia who had been immunized for the first time in 1993-94 and then in 1996-97. METHODS: In autumn 1996-97, 38 previously vaccinated hemophiliac patients were subcutaneously immunized with a single 0.5-mL dose of subunit influenza vaccine containing the following three virus strains: A/Singapore/6/86 (H1N1), A/Wuhan/359/95 (H3N2) and B/Beijing/184/93 (HB). Antibody response to influenza vaccine was measured before vaccination, 3 weeks after vaccination and 6 months after vaccination, by use of hemagglutinin- and neuraminidase-inhibition tests. To present the level of seroconversion, geometric mean titers of anti-influenza antibodies, mean fold increase, protection rate and conversion rate were determined. All results were compared with the control group of 23 healthy persons who had never been vaccinated against influenza and for whom the same serologic tests were carried out as for the vaccinated group. RESULTS: Three weeks after immunization, antihemagglutinin antibody levels were 3.9-10.9 times higher than before vaccination, but the highest mean fold increase values were recorded 6 months after vaccination, ranging from 8.4 to 28.6. In the case of neuraminidase, mean fold increases of antibodies reached values of 3.6-12.3 three weeks after vaccination and 7.1-29.1 six months after vaccination. The highest proportion of subjects protected was observed 6 months after immunization and ranged from 76.3% to 97.4%, compared to 52.6-60.5% 3 weeks after vaccination. Similar values were obtained for conversion rate: 71.1-86.8% 6 months after vaccination, in comparison with 39.5-42.1% 3 weeks after immunization. CONCLUSIONS: All data obtained in the present study indicate a significant immune response to subunit trivalent influenza vaccine in patients suffering from hemophilia; this is additionally confirmed by the fact that none of the vaccinated children were infected with the influenza virus and no serious adverse reactions were observed after administration of the vaccine. PMID- 11864249 TI - Effect of clindamycin given alone or with Lactobacillus delbrueckii and Streptococcus thermophilus on 7alpha-dehydroxylation of bile acids in rats. AB - OBJECTIVES: To evaluate the influence of clindamycin, given alone or with Lactobacillus delbrueckii and Streptococcus thermophilus to conventional rats, on 7alpha-dehydroxylation of cholic acid to deoxycholic acid. METHODS: The presence of deoxycholic acid was determined by gas-liquid chromatography. The fecal concentration of clindamycin was determined on PDM antibiotic sensitivity medium. Colony-forming units of L. delbrueckii and S. thermophilus were counted on lactic acid bacteria agar, and their ability to deconjugate was determined by thin-layer chromatography. RESULTS: Clindamycin significantly reduced the formation of deoxycholic acid, while the administration of lactobacilli-streptococci at the same time significantly reduced the effect of the antibiotic (p<0.05). CONCLUSIONS: Clindamycin reduced the formation of deoxycholic acid in rats and this effect was diminished by concomitant administration of L. delbrueckii and S. thermophilus. As the given microbes could not 7alpha-dehydroxylate, it seems reasonable to assume that they stimulate microbial species already present in the intestine. PMID- 11864250 TI - Isolated pulmonary valve endocarditis caused by Streptococcus bovis: case report and review. PMID- 11864251 TI - Comparative in vitro activity of alexomycin (U-82127) tested against Escherichia coli, Salmonella spp., and enterococci of animal and human origin. PMID- 11864252 TI - Sepsis and neutropenia induced by clozapine. PMID- 11864253 TI - European guidelines for diagnosis and management of patients with suspected herpes simplex encephalitis. PMID- 11864254 TI - A case of acute cholecystitis due to Aeromonas sobria and Hafnia alvei from northern Europe. PMID- 11864256 TI - The Second International Symposium on Infection Models in Antimicrobial Chemotherapy. PMID- 11864257 TI - Ethical considerations in the use of animal models in infection. PMID- 11864255 TI - Kawasaki disease and parvovirus B19 infection in an adult HIV-1-infected patient. PMID- 11864258 TI - The use of animal models in the investigation of the therapeutic role of cytokines in sepsis. PMID- 11864259 TI - Visna in sheep as a model for chemotherapy of lentiviral central nervous system infections. PMID- 11864260 TI - Antimicrobial susceptibility of Streptococcus pneumoniae from children attending day-care centers in a central Italian city. AB - OBJECTIVE: To undertake a survey of nasopharyngeal carriage of Streptococcus pneumoniae, which reflects strains causing infection, in 100 children under 3 years of age attending day-care centers in Frosinone, a city near Rome. METHODS: Fifty-three unique isolates of S. pneumoniae, isolated from 41 of the children tested, were tested for antimicrobial susceptibility to penicillin, cefotaxime, erythromycin, clindamycin, tetracycline, chloramphenicol and trimethoprim-- sulfamethoxazole. RESULTS: Resistance rates were as follows: penicillin, 20.7% (15% intermediate; 5.7% resistant); trimethoprim---sulfamethoxazole, 64.2%; erythromycin, 64.2%; clindamycin, 30.2%; tetracycline, 32.1%; and chloramphenicol, 3.8%. Except for three intermediate strains, all strains were susceptible to cefotaxime. Only five strains were susceptible to all of the antibiotics tested. An unusual finding of this study was that 23 of the 34 erythromycin-resistant strains were penicillin susceptible, whereas erythromycin resistant strains found in other countries are predominantly penicillin resistant as well. In addition, 18 of the 34 erythromycin-resistant strains were susceptible to clindamycin. Serogroups 6, 14, 19 and 23 accounted for 84.9% of the isolates. CONCLUSIONS: These data show that carriage of antibiotic-resistant pneumococci in children under 3 years of age is high in Frosinone, Italy. Information on resistance rates in pneumococcal disease in different age groups and on prevalence of drug resistance in other parts of the country is urgently needed. PMID- 11864261 TI - Incidence, epidemiology and evolution of reduced susceptibility to ciprofloxacin in Neisseria gonorrhoeae in Korea. AB - OBJECTIVE: To verify the decrease of susceptibility to ciprofloxacin in Neisseria gonorrhoeae, determine the size of the recently reported new beta-lactamase plasmid and explain the high prevalence of penicillinase-producing Neisseria gonorrhoeae (PPNG). METHODS: Gonococci were isolated from prostitutes in Korea. Antimicrobial susceptibility was tested by NCCLS disk diffusion and agar dilution methods. Plasmid was isolated by an alkaline lysis method. Patterns of Nhel digested genomic DNA were compared after pulsed-field gel electrophoresis (PFGE). RESULTS: The minimum inhibitory concentration of ciprofloxacin for 50% of the isolates rose from 0.015 mg/L in 1993 to 0.12 mg/L in 1996. The proportion of PPNG remained at 70% or over during the 5-year period. The size of a novel beta lactamase plasmid, first reported in 1994, was determined to be approximately 3.2 MDa, and 48% of the PPNG isolates contained it. Twelve of 50 isolates had the same PFGE pattern and nine others another pattern. CONCLUSION: The rapid decrease of fluoroquinolone-susceptible gonococci suggests that in the near future the drug may become less useful for gonorrhea treatment. The new 3.2-MDa plasmid may have been introduced as a result of the recent increase in overseas travel. The PFGE pattern suggests that high prevalence of PPNG may be due to dissemination of a few resistant clones among the high-risk groups. PMID- 11864262 TI - Antibodies against Helicobacter pylori in feces and saliva before and after eradication therapy. AB - OBJECTIVE: To determine specific antibody levels against Helicobacter pylori antigens in stool and saliva specimens from H. pylori-infected subjects and seronegative controls. METHODS: Antibody levels against a total membrane preparation (MP) and purified flagellin from H. pylori were measured by enzyme linked immunosorbent assays (ELISAs) in fecal extracts (FEs) and sera from 21 H. pylori-infected and 17 non-infected adult Swedish volunteers; specific antibody levels in saliva (IgG and IgA titers) were also measured in the majority of them. Follow-up specimens after successful eradication of H. pylori were collected after 5--7 weeks (all specimens) and 5 months (FEs and sera). RESULTS: Geometric mean specific IgA antibody levels, adjusted to total IgA concentration, in FEs were significantly higher (p<0.05) in the H. pylori-infected than in the control group against MPs (p<0.05) and flagelin (p<0.05). The total IgA concentrations in FEs from infected and seronegative controls were comparable and the IgA was mainly of the secretory type. Saliva specimens from H. pylori-infected subjects had higher ELISA IgA than IgG titers against MP. Anti-MP antibodies of both isotypes were higher in the infected than in the non-infected group (p<0.05 for both isotypes). The total IgA levels were also significantly (p<0.05) higher in saliva from the H. pylori-infected subjects than in saliva from the controls (geometric mean 292 mg/L and 173 mg/L, respectively). Eradication therapy resulted in a more rapid decline in antibody levels in FEs than in corresponding sera or saliva specimens. CONCLUSIONS: Specific antibodies against H. pylori MP were detected in FEs as well as in saliva from most of the infected volunteers, the mean antibody levels being significantly higher in specimens from the H. pylori-infected subjects than from seronegative controls. The results suggest that analyses of antibody levels in FEs may be used for follow-up of patients early (5--7 weeks) after onset of successful eradication therapy. PMID- 11864263 TI - High prevalence of GB virus C/hepatitis G virus infection in different risk groups of HIV-infected patients. AB - OBJECTIVES: To investigate the prevalence of GB virus C (GBV-C)/hepatitis G virus (HGV) RNA and anti-E2 antibodies in different risk groups of HIV-infected patients compared to that in healthy blood donors, and to study the effects of possible interactions between HIV and GBV-C/HGV on the carrier state and hepatic changes. METHODS: Sera from 100 consecutive unselected HIV-infected outpatients and from 100 healthy blood donors were screened for GBV-C/HGV viremia and anti-E2 antibodies. Anti-E2 antibodies were detected using an immunoassay developed by Boehringer Mannheim according to the manufacturer's instructions. GBV-C/HGV RNA was extracted from sera and reverse transcribed. The resulting cDNA was amplified with a PCR developed in the laboratory with primers derived from the 5prime prime or minute noncoding region of the viral genome and detected with a specific capture probe. This procedure was validated by a French multicenter quality control group. RESULTS: Thirty-one of the 100 HIV-infected patients and 8% of the healthy blood donors displayed anti-E2 antibodies. Four HIV-infected patients and one healthy blood donor were found to be GBV-C/HGV viremic. When analyzed by risk factor for the acquisition of HIV, no differences in the prevalence of anti-E2 antibodies were found between intravenous drug users and homosexual and heterosexual patients. CONCLUSIONS: We found a high prevalence of GBV-C/HGV infection in the HIV-infected population, irrespective of the risk group factor for HIV infection, suggesting that the sexual route is as effective as the parenteral route for the acquisition of GBV-C/HGV. No biological alteration could be attributed to GBV-C/HGV, even in the viremic patients. HIV-infected patients were able to clear GBV-C/HGV viremia and to mount a humoral immune response. PMID- 11864264 TI - Water-borne Campylobacter jejuni infection in a Danish town---a 6-week continuous source outbreak. AB - OBJECTIVE: To determine the cause and characteristics of illness of a Campylobacter jejuni outbreak in Denmark in 1995--96. METHODS: A retrospective follow-up study was designed for culture-confirmed cases and for residents without a bacteriologic diagnosis. Stored clinical and environmental isolates were analyzed by serotyping and genotyping with restriction endonuclease analysis (REA), pulsed-field gel electrophoresis (PFGE) and ribotyping. RESULTS: Campylobacter jejuni was isolated from 110 residents and visitors to the area. However, an estimate based on a telephone survey indicated that some 2400 people were affected by the outbreak. Water samples obtained from the community waterworks contained Campylobacter jejuni serotype O2, the same serotype as in all but one of the 30 stored isolates from the outbreak. The water and clinical isolates also showed the same DNA profile, except for the single strain showing the distinct serotype. The contamination of the water supply was traced back to contamination of ground water due to a break in a sewage pipe. CONCLUSIONS: A retrospective and demographic epidemiologic investigation of both culture confirmed and non-culture-confirmed cases in the town combined with typing of the isolates was crucial in defining the extent and cause of the outbreak. PMID- 11864265 TI - In vitro activity of vancomycin combined with rifampin, amikacin, ciprofloxacin or imipenem against methicillin-resistant and methicillin-susceptible Staphylococcus aureus. PMID- 11864266 TI - Group A streptococcal contact within schools over a 1-year period. PMID- 11864267 TI - Rapid emergence of meningococci with reduced susceptibility to penicillin in France: the need for vigilance in meningitis treatment. PMID- 11864268 TI - Typing strains of Chlamydia pneumoniae by amplified fragment length polymorphism typing. PMID- 11864269 TI - Legionella pneumophila: persistent positive sputum culture in an HIV-infected patient. PMID- 11864270 TI - Escherichia coli 0157 infection masquerading as 'rectal bleeding': a further problem for infection control. PMID- 11864271 TI - Lack of efficacy of ozone therapy in HIV infection. PMID- 11864272 TI - Lessons learned from the epidemiologic and microbiological study of a community outbreak of campylobacteriosis: sewage in the water, diarrhea in the community. PMID- 11864273 TI - The Yop virulon of Yersinia: a bacterial weapon to kill host cells. PMID- 11864274 TI - GB virus C/hepatitis G virus--- is it a novel human 'hepatitis' virus? PMID- 11864275 TI - Prevalence of diarrheagenic Escherichia coli strains detected by PCR in patients with travelers' diarrhea. AB - OBJECTIVE: To determine the prevalence of the different categories of diarrheagenic Escherichia coli, enterotoxigenic E. coli (ETEC), enteroinvasive E. coli (EIEC), verotoxin-producing E. coli (VTEC), enteroaggregative E. coli (EAggEC), diffusely adherent E. coli (DAEC), and enteropathogenic E. coli (EPEC), associated with travelers' diarrhea. METHODS: Stool specimens from 350 patients with travelers' diarrhea were collected between 1994 and 1996. The virulence factors of the diarrheagenic E. coli isolated were detected by PCR technique, in subcultures of single colonies of all morphotypes of E. coli observed in culture on MacConkey agar. RESULTS: ETEC (15.7%), EAggEC (13.4%) and DAEC (9.14%) are significantly more prevalent than EIEC (3.4%), EPEC (2.86%) and VTEC (0.86%) (p<0.05; z-test). The prevalence of ETEC and EAggEC was similar in all geographic areas visited. CONCLUSIONS: PCR is a rapid and specific technique to use in the identification of the different categories of diarrheagenic E. coli and greatly increases the yield of potential enteropathogens from cases of travelers' diarrhea. Not only ETEC but also EAggEC and DAEC strains play a major role in the etiology of travelers' diarrhea, whereas EIEC, EPEC, and VTEC strains play a minor role, leading to the question of whether it is necessary to search routinely for these three types of E. coli in diagnostic laboratories. PMID- 11864276 TI - Slime-producing properties of coagulase-negative staphylococci isolated from blood cultures. AB - OBJECTIVE: To evaluate five methods for the determination of slime-producing properties in coagulase-negative staphylococci (CNS). METHODS: One hundred and sixty-two strains of CNS considered as 'contaminants' and 162 strains associated with 'bacteremia' were tested with the tube test with tryptic soy broth, the tube test with brain---heart infusion broth supplemented with 5% sucrose, the Congo red agar method, and the microtiter-plate test with trypan blue and crystal violet, both with tryptic soy broth. RESULTS: Of the 324 strains tested, 188 were negative and 58 were positive with all methods. The remaining 78 strains were positive with one or more methods. CONCLUSIONS: There was a significant difference (p<0.001) in slime production between 162 strains of CNS pertaining to 'bacteremia' and 162 strains considered as 'contaminants', with 84 (51.8%) and 52 (32.8%) positive, respectively. The slime-producing strains were significantly more resistant (p<0.001) to cloxacillin, tobramycin, gentamicin, trimethoprim, erythromycin and ciprofloxacin. PMID- 11864277 TI - Serotyping and antibiotic resistance of Streptococcus pneumoniae isolated from pediatric infections in central Greece. AB - OBJECTIVE: To determine the distribution of serogroups/serotypes and antibiotic resistance pattern of Streptococcus pneumoniae isolated from pediatric infections in central Greece. METHODS: In total, 306 S. pneumoniae strains isolated from children, aged from 18 days to 14 years (median 18 months), during a 21-month period, from different specimen sources, were studied. Susceptibility testing was carried out by the Kirby---Bauer method and by the Etest, and serotyping by the Quellung reaction. RESULTS: Of the S. pneumoniae isolates, 3.9% were highly resistant to penicillin (PR), while 17.6% were intermediately resistant (IPR). PR and IPR isolates were found to be, in general, more resistant to other antibiotics than penicillin-susceptible isolates. The PR and IPR isolates belonged to the serogroup/serotypes 19, 23, 9, 6 and 14 (in descending order of frequency). The penicillin-susceptible isolates belonged to 20 different groups/serotypes, the most common being 19, 6, 14, 9, 3, 23 and 1 (in descending order of frequency). Serogroup 23 was often found to be multiresistant. CONCLUSIONS: Resistance to penicillin in S. pneumoniae isolates is relatively low and differs according to the specimen type. All the pneumococcal serogroups/serotypes isolated from the children were found to be included in the 23-valent polysaccharide vaccine. Most of the children with a pneumococcal infection, however, were less than 2 years old and could not be protected by the existing vaccine. PMID- 11864278 TI - Early bacterial and inflammatory responses to antibiotic therapy in a model of polymicrobial peritonitis in rats. AB - OBJECTIVE: To assess the consequences of different more or less selective treatments on the microbiological and inflammatory responses within the peritoneum. METHODS: The early effects of various antibiotic regimens were evaluated in a model of polymicrobial peritonitis with specifically prepared organisms. Six regimens (amoxycillin plus gentamicin, pefloxacin, ornidazole, pefloxacin plus ornidazole, imipenem and imipenem plus gentamicin) were evaluated at 24 h and 3 days in a non-fatal model of peritonitis in rats achieved by implantation of a capsule containing Escherichia coli, Bacteroides fragilis and Enterococcus faecalis. RESULTS: Therapies that disregarded several organisms were associated with persistence of the strains and an increased peritoneal inflammatory response within the peritoneum. In contrast, therapies active against Enterobacteriaceae and anaerobes were associated with decreases of all the inoculated organisms and a smaller inflammatory response. CONCLUSION: Therapies that disregarded the microorganisms implicated in peritoneal infection are associated with delayed bacterial eradication. The persistence of these organisms within the peritoneal fluid might be involved in prolonged peritoneal inflammation. Although it disregards enterococci, the standard therapy, represented by therapy against Enterobacteriaceae and anaerobes, demonstrates satisfactory effects towards all the inoculated organisms. This apparent contradiction could be related to mechanisms of bacterial synergy. PMID- 11864279 TI - Aspergillus species isolated from clinical specimens: suggested clinical and microbiological criteria to determine significance. AB - OBJECTIVE: To test a case-definition score for its usefulness in the diagnosis of pulmonary aspergillosis and to determine microbiological criteria useful in assessing the significance of isolating Aspergillus species from lower respiratory tract specimens (LRTS). METHODS: With the exception of cystic fibrosis patients, all patients with Aspergillus isolated from LRTS were classified according to a clinical case-definition ratio as: (i) colonized patients; (ii) patients with isolates of undetermined significance; (iii) patients with proven or probable pulmonary aspergillosis. Retrospective review of pathology records led to the identification of additional patients with histologically compatible pulmonary aspergillosis. Microbiological variables were compared between aspergillosis and colonization groups. RESULTS: Seventy-six isolates of Aspergillus species were obtained from 73 patients. For patients with proven (n=2) or histologically compatible pulmonary aspergillosis (n=5), the case definition ratio was always higher than 0.3; no patient with proven disease was classified into the colonized group. Compared to patients considered to be colonized (n=26), patients with proven or probable pulmonary aspergillosis showed a significant difference in the total number of Aspergillus colonies cultured per episode. In addition, for bronchoalveolar lavage specimens, the mean number of Aspergillus colonies/agar plate was also significantly higher in the latter group. The sensitivity and specificity of Aspergillus spp. detection by culture of LRTS for proven or probable pulmonary aspergillosis was 35.7% and 70.4%, respectively. CONCLUSION: The case-definition ratio used in this study helps interpret the clinical significance of Aspergillus isolated from LRTS. Data for assessment should include the absolute number of colonies recovered from culture of any LRTS and, for bronchoalveolar lavage, the mean number of Aspergillus colonies per agar plate. PMID- 11864281 TI - Commentary to 'Community-acquired pneumonia due to Escherichia coli' by Marrie TJ et al. PMID- 11864280 TI - Community-acquired pneumonia due to Escherichia coli. AB - OBJECTIVE: To describe the features of community-acquired Escherichia coli pneumonia and to compare these patients with patients with pneumonia caused by other etiologic agents. PATIENTS AND METHODS: This prospective study was carried out at five medical institutions in three geographic locations---Pittsburgh, PA, Boston, MA and Halifax, NS. Pneumonia etiology was assigned, based on results of microbiological investigations, by a committee consisting of five investigators using rules established prior to the study. Demographic and clinical features and outcomes of patients with E. coli pneumonia were compared with those of pneumonia due to other microorganisms. RESULTS: Nineteen patients (9 (47.4%) blood culture positive) had pneumonia due to E. coli and 430 (69 (16.0%) blood culture positive) had pneumonia caused by other etiologic agents. E. coli was the second most common cause of bacteremic pneumonia. The E. coli patients were older, and more likely to be female, from a nursing home and confused compared with patients with pneumonia due to other microbial agents. They were more severely ill as measured by a validated pneumonia specific severity of illness scoring measure. Although there was no in-hospital mortality for the patients with E. coli pneumonia, the 90-day mortality was 21%. Thirty-two (7.4%) of the patients with pneumonia due to other agents died in hospital and the 90-day mortality rate was 13.5% (p NS). Eight of the 19 patients with E. coli pneumonia were admitted from a nursing home and an additional four patients (63.2%) were discharged to such a facility. In contrast, only 44 (10.2%) of the patients with pneumonia due to other agents were discharged to a nursing home (p<0.001). CONCLUSIONS: Patients diagnosed with E. coli pneumonia are frequently bacteremic. They are older than patients with pneumonia due to other etiologies, and more likely to be female, from a nursing home and severely ill. Despite the absence of in-hospital mortality, 21% of these patients died within 90 days of presentation. PMID- 11864283 TI - Activity of beta-lactam antimicrobial agents against clinical isolates of Bacteroides fragilis in Kuwait. PMID- 11864282 TI - Prevalence of antibiotic resistance in Helicobacter pylori strains in Poland. PMID- 11864284 TI - The bactericidal activity of gatifloxacin in plasma and urine [1]. PMID- 11864285 TI - Reading Haemophilus influenzae Etest macrolide and amoxycillin susceptibilities: inter-observer variation appears to be small. PMID- 11864286 TI - Cefpirome---vancomycin combination in methicillin-resistant Staphylococcus aureus infection. PMID- 11864287 TI - Molecular typing methods and their discriminatory power. PMID- 11864288 TI - Immunoenzymatic dot-blot test for the diagnosis of enteric fever caused by Salmonella typhi in an endemic area. AB - OBJECTIVE: To develop a quick, solid-phase immunoenzymatic test, dot blot, to diagnose typhoid fever by the detection of IgG and IgM antibodies against three Salmonella typhi antigens (lipopolysaccharide, crude and flagellar) and to compare the dot-blot test with an enzyme-linked immunosorbent assay (ELISA) using the same antigens and with the Widal agglutination test. METHODS: Blood culture was used as the definitive test. All tests were used for the study of the sera from three groups of individuals: 33 patients suffering typhoid fever, diagnosed by isolation of S. typhi in blood culture; 35 patients with other enterobacterial infections documented by culture; and 156 asymptomatic volunteers, all residents of the same endemic region (Uraba, Colombia). RESULTS: The best diagnostic efficiency was obtained by detecting IgG against a flagellar antigen dot blot, giving a sensitivity and specificity of 88%. Widal and ELISA tests showed lower diagnostic efficiencies. CONCLUSIONS: The dot-blot test may be useful to diagnose typhoid fever in rural areas lacking technological resources with which to carry out blood cultures or ELISA tests. The visual reading makes this test practical and cheap for these regions. PMID- 11864289 TI - Use of molecular typing to investigate bacterial translocation from the intestinal tract in malnourished children with Gram-negative bacteremia. AB - OBJECTIVE: To investigate whether translocation of intestinal Enterobacteriaceae was a likely source of bacteremia in children of 2 years or less hospitalized for fever and moderate malnutrition. METHODS: Blood and fecal cultures were obtained from 198 such children admitted in the Princess Basma Hospital (Irbid, Jordan). Enterobacteriaceae isolated from blood cultures were compared with those present in fecal cultures by genomic typing using pulsed-field gel electrophoresis. The characteristics of children whose blood and fecal strain typing patterns were not distinguishable, i.e. with documented translocation, were compared to those of children without documented translocation. RESULTS: Bacterial translocation was documented in 12 of 19 children (63%) with Enterobacteriaceae bacteremia for whom paired blood and fecal strains were available for genomic typing, and was thus the major source of these bacteremias. The clinical or biological signs of malnutrition were not significantly more pronounced in the bacteremic children with documented translocation than in those without. CONCLUSIONS: We demonstrated that bacterial translocation from the intestinal tract was the most common source of Gram-negative bacteremia in the population studied. PMID- 11864290 TI - Mechanisms of quinolone resistance in clinical isolates of Enterobacter cloacae. AB - OBJECTIVE: To study and evaluate changes in the gyrA gene and the outer-membrane protein patterns in relation to evolution of resistance against the quinolones in Enterobacter cloacae. METHODS: Strains expressing gyrA-mediated quinolone resistance become susceptible to quinolones upon insertion of the plasmid pNJR3 2. This plasmid (containing wild-type Escherichia coli quinolone-susceptible DNA gyrase A subunits) and pLA2917 (the vector) were introduced into 10 resistant or moderately susceptible clinical isolates of Enterobacter cloacae by conjugation. The transconjugants, the original isolates, the plasmid and the vector control were screened for susceptibility to ofloxacin, ciprofloxacin and sparfloxacin. Additionally, examinations of the outer-membrane proteins were performed. RESULTS: A reduction of MICs by a factor of 8--32 was found for the transconjugants of five Enterobacter cloacae isolates in the presence of the gene probe, suggesting that these isolates harbored mutations in gyrA. No discernible difference in the patterns of outer-membrane proteins of sensitive and resistant strains could be detected. CONCLUSIONS: It seems that changes in the target site such as alterations in gyrA are important factors leading to a change in the susceptibility of bacteria to the quinolones, whereas there were no evident changes in the outer-membrane proteins to account for evolution of resistance. PMID- 11864291 TI - Evaluation of the discriminatory power of pulsed-field gel electrophoresis and PCR fingerprinting for epidemiologic typing of Candida species. AB - OBJECTIVE: To evaluate the discriminatory power of genotyping methods (PCR fingerprinting and pulsed-field gel electrophoresis) validated for Candida albicans in other Candida species. METHODS: Molecular typing methods are increasingly being applied for studies where the interpretation of data essentially relies on the typing results rather than epidemiologic data. In this situation, the discriminatory power (ability to identify differences among epidemiologically unrelated strains) of the typing method is important in allowing one to draw valid conclusions. By applying PCR fingerprinting, electrophoretic karyotyping, and restriction fragment endonuclease analysis using standard restriction enzymes and primers proven to be useful in previous studies, we evaluated whether the use of multiple genotyping methods is sufficient to delineate known unrelated strains among seven Candida species. RESULTS: All three methods identified individual genotypes for each of the seven Candida species studied. However, optimal strain delineation required the combined use of all three typing methods and was observed only within the small number of C. albicans and C. tropicalis isolates tested in this study. CONCLUSION: Typing assays that are able to delineate a certain Candida species may not be used blindly for other species of that genus. Regarding the limited number of strains tested, further validation of the discriminative power of genotyping methods (including in C. tropicalis) should be done. PMID- 11864292 TI - Evaluation of the BacT/Alert and VITAL blood culture systems for the diagnosis of bacteremia. AB - OBJECTIVE: To evaluate the detection of bacterial growth in the BacT/Alert (Organon Teknika) and VITAL (bioMerieux) automated blood culture systems. METHODS: In accordance with the protocol of study, 1021 blood sample pairs for culture were obtained from adult patients admitted to the Emergency Room and Intensive Care Unit. RESULTS: In total, 139 (13.6%) clinically significant blood cultures were detected, of which 79 (56.8%) were detected by both systems, 48 (34.5%) only by BacT/Alert and 12 (8.6%) only by VITAL (P cent0.0001). The BacT/Alert system detected positive blood cultures more rapidly for all groups of microorganisms. The VITAL system showed six false-negative blood cultures, while the BacT/Alert system showed none (P50.03). There was no significant difference between the number of false-positive blood cultures detected by the two systems. CONCLUSIONS: In our study, overall the BacT/Alert system achieved a better recovery of microorganisms than the VITAL system. PMID- 11864294 TI - Infections in patients requiring ventilation in intensive care: a critique of the application of a new classification. PMID- 11864293 TI - Infections in patients requiring ventilation in intensive care: application of a new classification. AB - OBJECTIVE: To classify infections according to the carrier state determined by surveillance cultures of throat and rectum, rather than by the traditional criterion of the time of onset after admission. METHODS: An observational cohort study of 3 months' duration was performed in a mixed medical---surgical intensive care unit (ICU) in a district general hospital of a subset of patients requiring mechanical ventilation for 3 days. Surveillance cultures from throat and rectum were obtained on admission to the ICU and then twice weekly to distinguish carriage of potentially pathogenic microorganisms (PPM) brought in by the patient from microorganisms acquired during the ICU stay. RESULTS: Out of the total population of 104 patients, 21 patients were enrolled over 3 months. Eight patients (38%) developed 12 infections, half of which were of primary endogenous pathogenesis and caused by Haemophilus influenzae, Candida albicans and Pseudomonas aeruginosa carried by the patients on admission. The remaining six were of secondary endogenous pathogenesis and caused by Acinetobacter baumannii and Pseudomonas aeruginosa acquired in the unit. CONCLUSIONS: Traditional classifications of hospital infection are challenged. If the traditional 48-h cut off point was used, then 9 of 12 cases (75%) of infection would have been classified as nosocomial, whereas using the method based on the carrier state, 50% of all infections were caused by microorganisms carried by the patient on admission to the ICU. Moreover, we believe that the distinction between primary endogenous, secondary endogenous and exogenous is valid because these three types of infection each require different control methods. PMID- 11864295 TI - Reply to Dr Spencer. PMID- 11864296 TI - Continuing Medical Education --- A New Paradigm. PMID- 11864297 TI - The Professors' Debate --- How to manage primary HIV infection. PMID- 11864298 TI - Severe sepsis due to Mycobacterium tuberculosis following extracorporeal shock wave lithotripsy. PMID- 11864299 TI - Bacteremia due to Roseomonas spp. PMID- 11864300 TI - Methylobacterium bacteremia in AIDS. PMID- 11864301 TI - Acute cholecystitis and bacteremia caused by Kluyvera ascorbata in a cirrhotic patient. PMID- 11864303 TI - Comparison of the effect of two decontamination procedures on the detection of Mycobacterium tuberculosis complex in respiratory specimens by a target-amplified test. PMID- 11864302 TI - Haemophilus influenzae type b endocarditis in a woman with mitral valve prolapse. PMID- 11864304 TI - Colony-stimulating factors and antibiotics---a new prospect in treating infectious diseases? PMID- 11864305 TI - Characteristics of pathogenic Neisseria meningitidis in Moscow: prevalence of 'non-European' strains. AB - OBJECTIVE: To investigate the distribution of serogroups, serotypes and subtypes, and susceptibility to antibiotics, of 75 strains isolated from patients with systemic meningococcal disease in Moscow in 1993--95. RESULTS: In contrast to the situation in most European countries, 21% of group A strains were found. Sixty nine per cent of the strains were non-serotypeable using the current panel of antibodies, and 21% of strains were non-subtypeable. Twenty-nine different serotype---subtype combinations were found among 69 strains of group A, B and C. No combination predominated clearly; relatively more frequent strains had the formulae B:NT:P1.2, A:4:P1.5, 10 and C:4:P1.10. Recently, such strains have been very rare in western Europe; in contrast, the strains predominating in western Europe were not found in Moscow. All strains were sensitive to penicillin, chloramphenicol and rifampicin. CONCLUSIONS: Moscow strains of Neisseria meningitidis demonstrated a substantial diversity of serotypes and subtypes that probably corresponded to a post-epidemic situation in Russia. The obvious difference in circulating strains and presumably in immunity of populations in western Europe and Russia increases the probability of mutual exchange of pathogenic strains and stresses the need for group B vaccine protecting from both western and eastern European variants of meningococci. PMID- 11864306 TI - Production of tumor necrosis factor-alpha and interleukin-6 in whole blood stimulated by live Gram-negative and Gram-positive bacteria. AB - OBJECTIVE: To investigate the production of tumor necrosis factor-alpha (TNF alpha) and interleukin-6 (IL-6) induced by live Gram-negative and Gram-positive bacteria in whole blood in vitro. METHODS: In all, 49 different isolates were studied. Each of the 49 different isolates was incubated for 4 h with whole blood at a ratio of one monocyte per 1--5 bacteria. Plasma was then separated and frozen, and the concentrations of TNF-alpha and IL-6 were measured by enzyme immunoassays. RESULTS: There was a positive correlation between TNF-alpha and IL 6 values, r=0.9. Gram-negative bacteria induced higher levels of both TNF-alpha and IL-6 than Gram-positive bacteria. Group G streptococci (GGS) induced higher levels of TNF-alpha than Streptococcus pneumoniae, Staphylococcus aureus, Staphylococcus epidermidis and group A streptococci (GAS). Klebsiella pneumoniae induced higher levels of TNF-alpha than Haemophilus influenzae, Escherichia coli and Neisseria meningitidis. GGS induced higher levels of IL-6 than Staphylococcus epidermidis, Staphylococcus aureus and GAS. When the relative amounts of cytokine induced by the strains were compared to serum concentrations measured on admission in patients with bacteremia caused by the same bacterial isolates there was no significant correlation. CONCLUSION: Species- and strain-related differences in cytokine-inducing properties were found which may have significance in clinical infections. PMID- 11864307 TI - Sparfloxacin as alternative treatment to standard therapy for community-acquired bacteremic pneumococcal pneumonia. AB - OBJECTIVE: To assess the effectiveness of a single daily dose of sparfloxacin in comparison with standard antibacterial therapy for the treatment of pneumococcal bacteremic community-acquired pneumonia (CAP). METHODS: The results were analyzed of four comparative trials in CAP, in which 107 adult patients with CAP confirmed by blood cultures positive for Streptococcus pneumoniae were included. Sparfloxacin was given at a loading dose of 400 mg followed by 200 mg daily. Comparator drugs included amoxycillin 3 g/day, amoxycillin/clavulanate 1.5/0.375 g/day and erythromycin 2 g/day. Dosing was for 7--14 days (mean 10 days). Success was determined by a combination of clinical and microbiological assessment and radiologic changes. RESULTS: Sparfloxacin was as effective as the comparator drugs, with an overall success rate of 80% at the end of treatment (comparators 78%), and a 79% success rate at follow-up (76% for comparators). There were no pneumococcal isolates resistant to sparfloxacin, but eight of 56 were either resistant (four) or had reduced susceptibility to penicillin G, and two strains were resistant to erythromycin. Tolerance to sparfloxacin was good, with fewer patients reporting drug-related adverse events (15.8%) than with the comparator drugs (33.3%). CONCLUSIONS: This analysis suggests that sparfloxacin would be an alternative candidate for empirical therapy in moderately severe CAP. PMID- 11864308 TI - Application of polymerase chain reaction for detection of Legionella pneumophila in serum samples. AB - OBJECTIVE: To apply the polymerase chain reaction (PCR) to serum samples for the rapid diagnosis of Legionnaire's disease using the L5SL9 and L5SR93 primers designed to generate a 104-base-pair (bp) fragment from the 5S RNA gene of Legionella spp. The amplified product was detected by electrophoresis and by hybridization with the L5S-1-specific probe. METHODS: Single specimens of serum obtained from 24 patients with confirmed legionellosis, at different stages of their disease, were tested by PCR. Additionally, 10 serum samples from patients with no clinical symptoms of pneumonia and 10 samples from patients suffering from pneumonia caused by Mycoplasma pneumoniae, Coxiella burnetii or Chlamydia psittaci were also tested as controls in order to determine the specificity of the method. RESULTS: Of the 24 examined serum samples, the amplified products from 12 hybridized with the L5S-1 probe (sensitivity 50%). None of the negative controls was positive after PCR. No correlation was found between the day of illness and the positivity in the test. CONCLUSIONS: The PCR technique could be applied as a diagnostic tool for the rapid diagnosis of legionellosis in serum samples after modification, mainly to improve its sensitivity. PMID- 11864309 TI - Evaluation of arbitrarily primed polymerase chain reaction analysis for typing Legionella pneumophila. AB - OBJECTIVE: To evaluate the performance of arbitrarily primed polymerase chain reaction (AP-PCR) analysis in epidemiologic typing of Legionella pneumophila. METHODS: Sixty-two isolates of L. pneumophila of serogroups 1, 3, 6 and 10, including epidemiologically related and unrelated isolates, were analyzed by AP PCR using the primer BG2. Twenty-six of the serogroup 1 isolates were typed by pulsed-field gel electrophoresis (PFGE). RESULTS: AP-PCR analysis showed 98% typeability and complete reproducibility. A majority of unrelated isolates of each serogroup could be distinguished (discrimination index: 92%). Clinical isolates showed AP-PCR patterns indistinguishable from those of the isolates of the related environmental source. PFGE and AP-PCR results were in agreement for 88% of isolates. CONCLUSIONS: Single-primer AP-PCR analysis can be used as a simple and reproducible screening method for typing L. pneumophila strains of different serogroups. PMID- 11864311 TI - Schistosoma mansoni myelitis in two patients who had traveled to West Africa. PMID- 11864310 TI - Illustrated continuing medical education. PMID- 11864312 TI - Acrodermatitis chronica atrophicans and serologic confirmation of infection due to Borrelia afzelii and/or Borrelia garinii by immunoblot. PMID- 11864313 TI - High intraplatelet cGMP levels in human sepsis. PMID- 11864314 TI - Listeria monocytogenes septic arthritis in a natural joint: report of a case and review. PMID- 11864315 TI - Escherichia coli producing a cephamycinase (CMY-2) from a patient from the Libyan --Tunisian border region. PMID- 11864316 TI - Comparative activities of amoxycillin and 10 other oral drugs against penicillin susceptible and -resistant Streptococcus pneumoniae strains recently isolated in Italy. PMID- 11864317 TI - Vaginal flora changes and reproducibility of interpretation of Gram-stained vaginal smears. PMID- 11864318 TI - Malnutrition and infection. PMID- 11864319 TI - Do the new lipid formulations of Amphotericin B really work? PMID- 11864320 TI - Mycobacterium celatum as a cause of disseminated infection in an AIDS patient. PMID- 11864322 TI - European guidelines for diagnosis and management of patients with suspected herpes simplex encephalitis. PMID- 11864321 TI - 'Think like a bacterium': a helpful concept to prolong the antibiotic era? PMID- 11864323 TI - Antianaerobic activity of a cecropin---melittin peptide. AB - OBJECTIVE: Several small, 15-residue peptides that contain portions of the amino acid sequences of both cecropin A and melittin have previously been shown to have broad-spectrum antibacterial activities against aerobic microorganisms, with no undesirable hemolytic properties. It would also be useful to know what effect these hybrid peptides have on anaerobic bacteria. METHODS: The minimum inhibitory concentrations of one hybrid, CA(1--7)M(2--9)NH2, were compared with those of seven other antimicrobial agents against 111 clinical anaerobic strains; Bacteroides fragilis, 24 strains; other Bacteroides fragilis group, 14 strains; other Bacteroides species, 13 strains; Fusobacterium nucleatum, six strains; Clostridium difficile, 22 strains; Clostridium perfringens, 10 strains, Propionibacterium spp., nine strains; and anaerobic cocci, 13 strains. RESULTS: Ninety per cent of strains belonging to the B. fragilis group, fusobacteria, propionibacteria and peptostreptococci were inhibited by 4 mg/L CA(1--7)M(2- 9)NH2, and the antimicrobial activity was approximately in the same range as that of chloramphenicol. CONCLUSION: This investigation showed that the antimicrobial spectrum of this cecropin---melittin hybrid also includes anaerobic organisms. PMID- 11864324 TI - Pharmacodynamic properties of HMR 3004, a novel ketolide, on respiratory pathogens, enterococci and Bacteroides fragilis demonstrated by studies of time kill kinetics and postantibiotic effect. AB - OBJECTIVE: The pharmacodynamic properties of the novel ketolide (a new class of macrolide) antibiotic, HMR 3004, were investigated by studying time-kill kinetics and postantibiotic effect. METHODS: The time-kill kinetics were studied at two inocula against three strains each of Staphylococcus aureus, Haemophilus influenzae, Moraxella catarrhalis, Streptococcus pneumoniae, Streptococcus pyogenes, Enterococcus faecalis, Enterococcus faecium and Bacteroides fragilis. The postantibiotic effects of HMR 3004 were also investigated on these organisms at concentrations equivalent to 1, 4 and 10 x MIC. RESULTS: The time kill-kinetic data demonstrated that HMR 3004 is inoculum dependent and predominantly bacteriostatic being only slowly bactericidal at higher concentrations. HMR 3004 exhibited a significant postantibiotic effect with all strains studied, ranging from 1.9--6.2 h at 10 x MIC. CONCLUSIONS: The bacteriostatic activity and significant postantibiotic effect demonstrated by HMR 3004 are similar to those previously obtained with other macrolides. PMID- 11864325 TI - Compassionate use of amphotericin B lipid complex (Abelcet) in life-threatening fungal infections: report of 30 courses. AB - OBJECTIVE: To report a single-center experience of compassionate use of amphotericin B lipid complex (ABLC) in patients with proven or suspected fungal infection who were or would have been unable to tolerate conventional amphotericin B. METHODS: Twenty-eight patients receiving 30 courses of ABLC for 22 proven invasive mycosis episodes (11 aspergillosis, seven candidosis, four miscellaneous) and eight suspected episodes are described. Seven patients were given ABLC first-line therapy because of conditions precluding the use of amphotericin B deoxycholate (Am B). Twenty-one patients, initially given Am B, were shifted to ABLC because of failure in four, nephrotoxicity of AM B alone or in combination with another drug in 15, and acute side effects in two. The initial dose of ABLC was 5 mg/kg per day; this could be lowered to 3 mg/kg per day or transiently interrupted in cases of impairment of renal function. RESULTS: A mean cumulative dose of 6107 mg (660--16 050) was given over a mean duration of 22 days (4--49). Clinical response rate was 63% (14/22), with mycologic eradication in 37% (9/17) in proven infections. For proven aspergillosis, corresponding rates were 54% (6/11) and 20% (2/10), and in proven candidosis 71% (5/7) and 60% (3/5), respectively. Twenty-one courses were complicated by one or more side effects: fever and chills (11), impairment of renal function requiring a transient reduction of drug dosage (14), hypotension (1). However, for the whole group, creatinine clearance before and after 2, 4 and 6 weeks of treatment remained quite stable. CONCLUSIONS: ABLC, with its low toxicity, enabled us to treat patients who were or would have been unable to tolerate an efficacious dose of Am B. No conclusions about efficacy can be drawn from this small-size, compassionate study. Well-designed studies to compare efficacy and safety of conventional amphotericin B and the various lipidic formulations should be implemented. PMID- 11864326 TI - Resistance to amikacin and gentamicin among Gram-negative bloodstream isolates in a university hospital between 1989 and 1994. AB - OBJECTIVE: To characterize antimicrobial resistance patterns to amikacin (AN) and gentamicin (GM) among Gram-negative bloodstream isolates and to determine the possible relationship between use of AN and GM and the occurrence of antibiotic resistance during a 6-year period. METHODS: Standard media and techniques of isolation and identification were used. Antimicrobial susceptibility testing was performed with the disk diffusion method and API rapid ATB E strips. Data on consumption of aminoglycosides were collected by the central hospital pharmacy and were expressed as daily defined doses. RESULTS: One thousand nine hundred and four bloodstream isolates were tested for AN and GM susceptibility between 1989 and 1994. Activities of AN and GM remained high during the study period against most isolates of Gram-negative bacteria. No relationship could be observed between the use of AN/GM and the rate of AN/GM resistance. Nosocomial Gram negative bloodstream isolates showed a higher degree of resistance towards both AN (3.9% of all nosocomial isolates) and GM (7.9%) than community-acquired isolates (1.8% toward AN and 3.1% towards GM, respectively). There was a significant increase (P=0.004) in the risk of GM resistance in patients with nosocomial Gram-negative bacteremia detected more than 14 days after admission. The proportion of GM-susceptible Pseudomonas aeruginosa isolates decreased linearly from 97% for infections acquired between day 3 and day 10 following admission to 80% for bacteremia developing 30 days or more after admission (P=0.008). CONCLUSIONS: AN and GM remain highly active antimicrobial drugs for treatment of GNB in times of growing resistance to cephalosporins and fluoroquinolones. PMID- 11864327 TI - Interpretation of immunoblots for Lyme borreliosis using a semiquantitative approach. AB - OBJECTIVE: To test the performances of new Borrelia garinii immunoblots specific for Borrelia burgdorferi sensu lato with a selected panel of sera from patients with various clinical presentations of Lyme borreliosis. METHODS: In order to establish the sensitivity and the specificity of these immunoblots, we tested serum samples obtained from patients with early- and late-stage Lyme disease (erythema migrans n=35, neuroborreliosis n=61, acrodermatitis chronica atrophicans (ACA) n=27 and arthritis n=41), from patients with diagnoses and laboratory findings associated with serologic cross-reactivity to Lyme disease (syphilis n=12, Epstein-Barr infection n=9, autoimmune markers n=29) and from blood donors residing in regions of low and medium endemicity (n=80, n=100). RESULTS: The combined sensitivity (IgG and IgM) of the tests was 90% for patients with erythema migrans, 92% for neuroborreliosis, 96% for ACA and 100% for Lyme arthritis. The specificity of the IgG immunoblot was 94%, and that of the IgM immunoblot was 97%, taking into account the prevalence of borrelia antibodies in the overall population. Interpretation of these immunoblots is based on scores allocated to different specific borrelia antigens. CONCLUSIONS: The Western blot technology is extremely useful in dissecting the immune response to borrelia infections, which develops gradually over a period of weeks to years and which involves the appearance of IgM and IgG antibodies directed against a number of borrelia-associated proteins. PMID- 11864328 TI - Isolation and characterization of enteroaggregative Escherichia coli (EAggEC) by genotypic and phenotypic markers, isolated from diarrheal children in Congo. AB - OBJECTIVE: To determine the prevalence of enteroaggregative Escherichia coli (EAggEC) in African diarrheal children in Lwiro, Congo, to characterize EAggEC isolates by possible genotypic and phenotypic markers, and to evaluate the EAggEC probe pCVD432 in identifying EAggEC. METHODS: The Hep-2 cell adhesion assay and colony-blot hybridization assays were carried out for the identification of EAggEC. O:H serotyping, biotyping, antibiogram and plasmid-profile analysis were done. To detect the E. coli LT and ST, ELISA tests were used and, for VT, a vero cell assay was used. RESULTS: EAggEC strains were isolated from 56 out of 115 diarrheal children (48.7%): the organism was present alone and presumed to cause diarrhea in 22 (19.1%) cases. The rest of the cases were associated with two or more diarrheagenic E. coli strains. EAggEC strains were isolated from 25% of total diarrheal children (first day of isolation) and 8.86% of age-matched healthy individuals (p<0.03). This isolation rate was significantly higher than the one found for other diarrheagenic E. coli strains. In parallel, we evaluated the sensitivity and specificity of EAggEC probe pCVD432, and found that it had 56% sensitivity with 100% specificity compared with the Hep-2 cell test. EAggEC isolates were characterized by serotyping, biotyping, antibiotic resistance pattern, plasmid profiling and toxin production analysis. They did not produce any one of these classical toxins and nor did they relate to any particular serotypes. Plasmid analysis of the 79 EAggEC isolates (n=315) showed seven different profiles. Ten resistance patterns were identified and 34 strains were sensitive to all drugs. There was no association between plasmid profiles and antibiotic resistance patterns. All 16 classical E. coli biotypes were found in this small EAggEC population. CONCLUSIONS: EAggEC has been emerging as a cause of childhood diarrhea in African children in Congo. From the accumulated data it was found that there is a great heterogeneity in EAggEC populations. PMID- 11864329 TI - Illustrated continuing medical education. PMID- 11864330 TI - Lack of association between chronic hepatitis and hepatitis G virus infection in Senegal. PMID- 11864331 TI - Diagnosis of disseminated toxoplasmosis by PCR analysis of ascitic fluid in a patient with haematologic malignancy. PMID- 11864332 TI - Surveillance of meningococcal infections in Belgium. PMID- 11864333 TI - First case of Salmonella hirschfeldii (paratyphi C) infection of a prosthetic hip. PMID- 11864334 TI - Successful treatment of Salmonella aortitis with ciprofloxacin. PMID- 11864335 TI - Response to Ian Phillips and Anna King. PMID- 11864337 TI - Procalcitonin, a new diagnostic and prognostic marker for severe infections. PMID- 11864336 TI - International Symposium on Salmonella and Salmonellosis, 20--21 May 1997, Ploufragan, France. PMID- 11864338 TI - Frequency and molecular diversity of Pseudomonas aeruginosa upon admission and during hospitalization: a prospective epidemiologic study. AB - OBJECTIVE: To assess the molecular epidemiology and risk factors for Pseudomonas aeruginosa colonization and infection in hospitalized patients. METHODS: In a 1000-bed university hospital, newly admitted patients were assessed prospectively for colonization and infection with P. aeruginosa. Anal swabs were obtained upon admission and at discharge. Ribotyping was used for the typing of isolates. Epidemiologic and clinical data were recorded prospectively. Independent risk factors were assessed using multivariate analysis. RESULTS: The recovery rate of patients with P. aeruginosa from anal specimens on admission was 6.7% (42/628). Infection due to P. aeruginosa was observed in 20 of 628 (3.2%) patients, of whom 10 (1.6%) were already infected on admission. Independent risk factors for colonization/infection on admission were age, indwelling urinary catheter, the presence of wound and seropositivity for HIV. Independent risk factors for nosocomial infection were anal colonization on admission, alcoholism, indwelling urinary catheter and antibiotic treatment during hospitalization. Ribotyping revealed that 27 patients were colonized or infected with a unique ribotype, whereas 24 shared one or more ribotypes with other patients. Analysis of epidemiologic and molecular typing data revealed that transmission from patients to patients or from patients to environment was documented on only three occasions. CONCLUSIONS: A great many P. aeruginosa strains were isolated from patients and the environment, and most environmental strains were different from those recovered in patients, suggesting a low rate of hospital acquisition of P. aeruginosa from the environment. The main risk factors for hospital-acquired infection were detectable colonization on admission, antibiotic treatment and urinary catheter. PMID- 11864339 TI - Molecular epidemiology of quinolone resistance in Acinetobacter spp. AB - OBJECTIVE: To determine whether similar mutations to quinolone resistance in the gyrA subunit of DNA gyrase and the parC subunit of topoisomerase IV are occurring independently in genotypically unrelated clinical isolates of Acinetobacter spp., or whether worldwide clonal spread of particular resistant strains is occurring. METHODS: The genotypic relationships of 25 nosocomial isolates of Acinetobacter spp. from 15 locations in 11 different countries worldwide were examined by randomly amplified polymorphic DNA analysis. Quinolone resistance-determining regions of gyrA and parC were amplified by PCR and mutations were analyzed by restriction digestion with Hinfl and DNA sequencing. RESULTS: Twenty-four of the 25 Acinetobacter isolates were genotypically heterogeneous and 12 were resistant to both nalidixic acid and ciprofloxacin. Analysis of conserved gyrA and parC regions showed that all isolates with a ciprofloxacin MIC of 4 mg/L had a substitution of Ser83 with either Leu or Phe in the GyrA protein. Five of six isolates with ciprofloxacin MICs of 64 mg/L had additional substitutions of Ser80 with Leu in the ParC protein. CONCLUSIONS: Similar mutations to quinolone resistance, predominantly at codons 82--83 of gyrA, are occurring independently in genotypically distinct isolates of Acinetobacter spp. from different worldwide locations. Most isolates with high ciprofloxacin MICs also exhibited secondary mutations in parC at codons 79--80. PMID- 11864340 TI - Whole cell protein and partial 16S rRNA gene sequence analysis suggest the existence of a second Rothia species. AB - OBJECTIVE: To subject ten clinical isolates grouped together based on their biochemical and microbiological profile to further investigations aimed at correct species identification. METHODS: The 16S rRNA gene was partially sequenced using nested amplification. Whole cell protein analysis (SDS-PAGE) and cluster analysis were performed on the 10 strains and also for comparison on 31 reference strains. The API Coryne biochemical kit as well as API 20 Strep were used for analysis of the phenotypic diversity of the strains by use of computerized numerical identification procedures. Antibiotic susceptibility testing was performed using a standardized disk diffusion test. RESULTS: The 265- 556-bp-long 16S rRNA gene sequences of all 10 strains showed highest similarity to Rothia dentocariosa. Three strains showed complete identity between the sequences obtained and the sequence of the type strain of Rothia dentocariosa 16S rRNA gene (M59055), and the other seven ranged between 99.7% and 98.3% similarity. Detailed analysis of the sequences revealed a clustering of the strains into two groups. One group consisted of four isolates with the highest degrees of similarity with the reference strain (type I), while the members of another group (type II) showed differences in their nucleotide sequence at four distinct positions in the variable V7 region. T was replaced by C at position 597, C by T at position 608, T by C at position 610, and G by A at position 684 (position numbers according to reference sequence M59055, EMBL/GenBank). Whole cell protein analysis (SDS-PAGE) and cluster analysis also segregated the 10 Rothia dentocariosa strains into two different clusters, with one cluster containing all four strains belonging to 16S rRNA gene type I, and a second cluster containing all six strains belonging to 16S rRNA gene type II. CONCLUSIONS: Partial sequence data of the 16S rRNA gene as well as whole cell protein analysis showed a subdivision of the Rothia species into two groups, genomovar I (Rothia dentocariosa sensu stricto) and genomovar II, a possible new Rothia species. PMID- 11864341 TI - Bactericidal effects of levofloxacin in comparison with those of ciprofloxacin and sparfloxacin. AB - OBJECTIVE: To investigate and compare the in vitro activity of levofloxacin with the activities of ciprofloxacin and sparfloxacin. METHODS: The following experiments were performed: (1) comparative studies of the rate of killing by the three quinolones of different strains of Streptococcus pneumoniae at a concentration corresponding to the 1-h serum level following a 500-mg dose in humans; (2) comparative studies of the rate of killing by levofloxacin and ciprofloxacin of different strains of Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa at the same concentrations as above; (3) comparative studies of the rate of killing by levofloxacin at four different concentrations of reference and clinical strains of Streptococcus pneumoniae, Staphylococcus aureus, E. coli and P. aeruginosa. RESULTS: Levofloxacin exhibited statistically significantly higher bactericidal activity than sparfloxacin after 2 and/or 3 h against all strains of Streptococcus pneumoniae. Compared to ciprofloxacin, levofloxacin showed a statistically significantly higher bactericidal activity after 2 and/or 3 h against all strains of Streptococcus pneumoniae except the one resistant to both penicillin and cefotaxime. No differences in killing rate between levofloxacin and ciprofloxacin were seen against Staphylococcus aureus, E. coli and P. aeruginosa, with almost complete killing after 3 h of the P. aeruginosa strains and after 6 h for the E. coli strains. No concentration dependent killing was seen at concentrations above 4xMIC of levofloxacin against Staphyloccus aureus, E. coli and P. aeruginosa. CONCLUSION: Levofloxacin was shown to be active against both Gram-positive and Gram-negative bacteria. In terms of MIC values, ciprofloxacin was the most active drug against the Gram negative organisms, and sparfloxacin against the strains of Streptococcus pneumoniae, but levofloxacin exhibited a similar or even better bactericidal activity against the investigated strains compared with the other two fluoroquinolones when killing curves were compared. PMID- 11864342 TI - Prospective 7-year monitoring of MAC bacteremia in HIV-positive Swedish patients. AB - OBJECTIVE: To record the cumulative incidence of Mycobacterium avium complex (MAC) bacteremia among HIV-infected patients and to study colonization in relation to symptoms of infection. METHODS: In a prospective study, 61 patients with a CD4 count less-than-or-eq, slant200x106/L were followed by cultures from sputum, feces and blood every 3--6 months and for development of MAC bacteremia and clinical symptoms. The main end-points were MAC bacteremia and death. RESULTS: From the start in November 1989 to January 1997 about 34% had developed MAC bacteremia with a median follow-up of 22 months. At the time of positive blood cultures, all but one patient had symptoms consistent with disseminated MAC infection. Positive cultures from respiratory and gastrointestinal tract were recorded before MAC bacteremia in only four patients. All but one had symptoms at the time of positive blood culture. CONCLUSIONS: The incidence of MAC bacteremia was similar to figures in other studies. The presence of symptoms in close relation to positive blood cultures supports late colonization and late infection in HIV disease. Screening patients with samples from the respiratory and gastrointestinal tracts is not useful PMID- 11864343 TI - The critical relationship between compliance and the management of infectious diseases. PMID- 11864344 TI - International multicenter investigation of LB20304, a new fluoronaphthyridone. PMID- 11864345 TI - Significant bacteriuria in infants and young children and relation to bacterial species and pyuria. PMID- 11864346 TI - Relapsing erythematous desquamating disorder in a child, with the same toxin producing Staphylococcus aureus strain. PMID- 11864348 TI - Methods for the determination of susceptibility of bacteria to antimicrobial agents. Terminology. PMID- 11864347 TI - Enterococcus avium osteomyelitis. PMID- 11864349 TI - The use of chimeric bacterial and plant protein toxins for targeted chemotherapy. PMID- 11864350 TI - A multicenter evaluation of the Biotest legionella urinary antigen EIA. AB - OBJECTIVES: To undertake a multicenter study to evaluate the Biotest legionella urinary antigen enzyme immunoassay (EIA) performance against those EIAs already in use in 14 European laboratories. METHODS: Each laboratory examined urine specimens from appropriate patients using both their current assay and the Biotest EIA. Each examined: a standard panel of 12 coded urine samples (distributed by Biotest); a panel of 10 coded urine samples provided as part of a European external quality assurance (EQA) scheme; urine samples from patients with proven legionnaires' disease (LD); urine samples from patients with pneumonia of microbiologically proven cause other than LD; and urine samples submitted for routine examination. Thus, the performance of the Biotest assay (in comparison with current EIAs), its specificity and utility, and the inter laboratory agreement were assessed. RESULTS: Inter-laboratory agreement was excellent, with all participants obtaining the expected results for 20 of 22 coded urine specimens. Specificity, determined using 123 specimens from patients with infections of known etiology, was 100%. The Biotest EIA gave positive results in 86% of specimens which had been positive in the laboratories' current EIAs, and in 94.6% of those specimens which were positive for Legionella pneumophila serogroup 1. CONCLUSION: The Biotest EIA is simple to use and specific and the results obtained in different laboratories show excellent agreement. The assay compares well existing EIAs, at least for L. pneumophila serogroup 1 PMID- 11864351 TI - Cytokine response to staphylococcal exotoxins in Staphylococcus aureus septicemia. AB - OBJECTIVE: To investigate if exposure to exotoxins results in augmented serum cytokine response of patients with Staphylococcus aureus septicemia. METHODS: Serum samples and strains from 63 patients with S. aureus septicemia were collected in a prospective study. Toxin production by strains in vitro was determined by enzyme immunoassay (EIA) or reversed passive latex agglutination (RPLA). Antibodies against the toxins and cytokine levels in serum on admission were analyzed with EIA. RESULTS: Patients infected with strains producing staphylococcal enterotoxins (SEs) A, B, C and D and/or toxic shock syndrome toxin 1 (TSST-1) in vitro (n=37) showed higher serum TNF-alpha levels than those infected with non-toxigenic strains (p=0.04). A significant titer rise against the corresponding SE and/or TSST-1 produced by the isolate (14/35), indirectly reflecting exposure to the antigen, was associated with higher TNF-alpha concentrations on admission than in those without titer rise (p=0.03). Patients with low antibody titers against SE and/or TSST-1 on admission (19/37) were found to have higher serum TNF-alpha concentrations on admission than those with elevated antibody titers on admission (p=0.03). CONCLUSION: Patients infected with toxigenic S. aureus strains produce significantly higher levels of serum TNF alpha on admission compared to patients infected with non-toxigenic strains. PMID- 11864352 TI - Virtually all methicillin-resistant Staphylococcus aureus (MRSA) infections in the largest Portuguese teaching hospital are caused by two internationally spread multiresistant strains: the 'Iberian' and the 'Brazilian' clones of MRSA. AB - OBJECTIVE: To determine the nature (clonal type and antibiotic resistance pattern) of methicillin-resistant Staphylococcus aureus (MRSA) strains recovered from the largest teaching hospital in Portugal and to detect temporal trends in clonal types during three consecutive surveillance periods in 1992--93, 1994--95 and 1996. METHODS: MRSA strains were characterized by chromosomal SmaI macrorestriction patterns using pulsed-field gel electrophoresis (PFGE) and by DNA fingerprints---applied to ClaI digests---capable of probing two specific areas of the staphylococcal chromosome: (1) the vicinity of the mecA gene, and (2) the attachment site(s) and copy number of transposon Tn554. The combination of these methods can generate 'clonal types' useful for epidemiological tracking of MRSA strains. RESULTS: During the 1992--93 collection period, 65% of MRSA strains carried the mecA polymorph I, Tn554 pattern E and PFGE pattern A (I::E::A)---a clonal type that was used to define the 'Iberian MRSA', which is widely spread throughout southern Europe. The representation of this clone decreased to 42% in 1994--95 and to 20% in 1996. At the same time, a second multiresistant MRSA strain carrying mecA polymorph XI, Tn554 type B and PFGE pattern B (XI::B::B)---a clonal type characteristic of the so-called 'Brazilian MRSA'---increased from 5% in 1992--93 to 36% in 1994--95 and 29% in 1996. CONCLUSIONS: Throughout the four years of surveillance, the Iberian and Brazilian MRSA types and their single subtype variants together have been responsible for the overwhelming majority (close to 90%) of all MRSA infections in the largest teaching hospital of Portugal. The mechanism of epidemicity of these two multiresistant international MRSA clones remains to be elucidated. PMID- 11864353 TI - Nosocomial legionellosis outbreak over a three-year period: investigation and control. AB - OBJECTIVE: To investigate the epidemiologic relatedness of nosocomial infections due to Legionella pneumophila serogroup 1 diagnosed between 1992 and 1994 in six immunocompromised patients of the same hospital and to describe the measures which were developed to control the outbreak. METHODS: Legionella strains isolated from patients and from potable hot water were compared using three typing methods: monoclonal antibody analysis, arbitrarily primed PCR and ribotyping. RESULTS: Environmental investigations revealed the presence of high levels of L. pneumophila serogroup 1 in hot water. The typing methods gave concordant results for demonstrating (1) the persistence of an epidemic strain of L. pneumophila serogroup 1 in the major water distribution circuit of the hospital over a 3-year period, and (2) the identity between patients' and environmental strains. Five of the six patients were probably infected via aerosols of hot tap water following inappropriate therapeutic procedures. Repetitive heat flushings associated with regular bacteriologic surveillance allowed correct disinfection of the water distribution systems. Specific recommendations concerning aerosol delivery and oxygen therapy were implemented in order to prevent further nosocomial legionellosis. CONCLUSIONS: The same strain of L. pneumophila had been able to colonize the main water circuit of the hospital for at least 3 years; the relatedness between clinical and environmental strains was easily confirmed by the use of molecular markers. PMID- 11864354 TI - The bactericidal activity of gatifloxacin in plasma and urine. AB - OBJECTIVE: To investigate the bactericidal activity of gatifloxacin in serum and urine against relevant pathogens. METHODS: Serum and urine samples obtained in a single rising dose pharmacokinetic study were investigated for bactericidal activity. The doses employed were placebo, and 200, 400, 600 and 800 mg of gatifloxacin. RESULTS: The titers obtained reflected the dose and susceptibility of the pathogen. In serum a titer of greater-than-or-equal1:8 was observed for Escherichia coli for >24 h after 400 mg; for Staphylococcus aureus and Acinetobacter baumanii the titer was greater-than-or-equal1:8 for between 12--24 h after 600 mg; for Serratia marcescens it was greater-than-or-equal1:8 for >12 h following 800 mg; and for Streptococcus pneumoniae it was greater-than-or equal1:8 for 8 h following 800 mg. In urine, the titers were greater, and for Escherichia coli, Staphylococcus saprophyticus, Proteus mirabilis and Enterococcus faecalis the titer was >1:8 for 12--24 h after 200 mg and for 24--36 h after a 400-mg dose. Pseudomonas aeruginosa demonstrated a titer of >1:8 for 18 -24 h following 600 mg. CONCLUSIONS: These data suggest that gatifloxacin should be efficacious for a wide range of systemic and urinary tract infections. PMID- 11864355 TI - Inhibition of amplification of Chlamydia trachomatis plasmid DNA by the ligase chain reaction associated with female urines. AB - OBJECTIVE: To determine the rate of inhibition of ligase chain reaction (LCR) amplification of Chlamydia trachomatis plasmid DNA in urines from pregnant and non-pregnant women, to correlate inhibitors with urinary substances through urinalysis, and to explore methods of removing inhibitors from urine. METHODS: First-void urines (FVUs) from 287 non-pregnant and 101 pregnant women (15--40 years of age) in Hamilton, Canada were tested for urinary factors by urinalysis and for C. trachomatis by LCR. A sample of each urine was 'spiked' with a positive control containing approximately one infectious unit of C. trachomatis L2 serovar. RESULTS: Complete inhibition of DNA amplification was observed in nine of 287 (3.1%) urines from non-pregnant women and in one of 101 (1%) who were pregnant. Partial inhibition (> 20% reduction in signal) was observed in an additional five of 388 (1.3%) urines. Nine urines contained C. trachomatis plasmid DNA, but none of these contained inhibitors of LCR. By multivariate analysis, the only substances in urine which were significantly and independently related to LCR inhibition were nitrites. Holding the urines at 4C or minus sign70C overnight reduced the numbers which were inhibitory by approximately 50%. Diluting the urines 1:10 or extracting with phenol/chloroform removed all inhibition and retained amplifiable DNA. CONCLUSIONS: We conclude that urinary inhibitors of C. trachomatis DNA amplification by LCR are minimal and appear to be related to the presence of nitrites. We anticipate that most would disappear in transit or during processing and that a positive control for every urine is not warranted, considering the added cost and work associated with duplicate testing. PMID- 11864357 TI - Hydatid disease. PMID- 11864356 TI - The detection of mycobacteria in blood cultures using the Bactec system: 6 weeks versus 12 weeks of incubation? routine terminal Ziel-Neelsen? PMID- 11864358 TI - Antimicrobial resistance in Listeria monocytogenes from humans and food in the UK, 1967--96. PMID- 11864359 TI - Failure to detect Helicobacter pylori in vaginal secretions. PMID- 11864360 TI - Enteric infections due to Escherichia coli. PMID- 11864362 TI - Reply from the author. PMID- 11864361 TI - Response to Drs Pierard and Lauwers. PMID- 11864366 TI - Rosetta Stone proteins: "chance and necessity"? PMID- 11864365 TI - The past, present and future of genome-wide re-annotation. AB - Annotation, the process by which structural or functional information is inferred for genes or proteins, is crucial for obtaining value from genome sequences. We define the process of annotating a previously annotated genome sequence as 're annotation', and examine the strengths and weaknesses of current manual and automatic genome-wide re-annotation approaches. PMID- 11864367 TI - Structural genomics: a new era for pharmaceutical research. PMID- 11864368 TI - 2001: a mouse genome odyssey. AB - A report on the 15th International Mouse Genome Conference, Edinburgh, UK, 21-24 October 2001. PMID- 11864369 TI - Evaluation of differential gene expression during behavioral development in the honeybee using microarrays and northern blots. AB - BACKGROUND: The honeybee (Apis mellifera) has been used with great success in a variety of behavioral studies. The lack of genomic tools in this species has, however, hampered efforts to provide genome-based explanations for behavioral data. We have combined the power of DNA arrays and the availability of distinct behavioral stages in honeybees to explore the dynamics of gene expression during adult development in this insect. In addition, we used caffeine treatment, a procedure that accelerates learning abilities in honeybees, to examine changes in gene expression underlying drug-induced behavioral modifications. RESULTS: Spotted microarrays containing several thousand cDNAs were interrogated with RNAs extracted from newly emerged worker bees, experienced foragers and caffeine treated bees. Thirty-six differentially expressed cDNAs were verified by northern blot hybridization and characterized in silico by sequencing and database searches. Experienced foragers overexpressed royal jelly proteins, a putative imaginal disc growth factor, a transcriptional regulator (Stck) and several enzymes, including alpha-glucosidases, aminopeptidases and glucose dehydrogenase. Naive workers showed increased expression of members of the SPARC and lectin families, heat-shock cognate proteins and several proteins related to RNA translation and mitochondrial function. A number of novel genes overexpressed in both naive and experienced bees, and genes induced by caffeine, have also been identified. CONCLUSIONS: We have shown the usefulness of this transcriptome-based approach for gene discovery, in particular in the context of the efficacy of drug treatment, in a model organism in which routine genetic techniques cannot be applied easily. PMID- 11864370 TI - Selection in the evolution of gene duplications. AB - BACKGROUND: Gene duplications have a major role in the evolution of new biological functions. Theoretical studies often assume that a duplication per se is selectively neutral and that, following a duplication, one of the gene copies is freed from purifying (stabilizing) selection, which creates the potential for evolution of a new function. RESULTS: In search of systematic evidence of accelerated evolution after duplication, we used data from 26 bacterial, six archaeal, and seven eukaryotic genomes to compare the mode and strength of selection acting on recently duplicated genes (paralogs) and on similarly diverged, unduplicated orthologous genes in different species. We find that the ratio of nonsynonymous to synonymous substitutions (Kn/Ks) in most paralogous pairs is <<1 and that paralogs typically evolve at similar rates, without significant asymmetry, indicating that both paralogs produced by a duplication are subject to purifying selection. This selection is, however, substantially weaker than the purifying selection affecting unduplicated orthologs that have diverged to the same extent as the analyzed paralogs. Most of the recently duplicated genes appear to be involved in various forms of environmental response; in particular, many of them encode membrane and secreted proteins. CONCLUSIONS: The results of this analysis indicate that recently duplicated paralogs evolve faster than orthologs with the same level of divergence and similar functions, but apparently do not experience a phase of neutral evolution. We hypothesize that gene duplications that persist in an evolving lineage are beneficial from the time of their origin, due primarily to a protein dosage effect in response to variable environmental conditions; duplications are likely to give rise to new functions at a later phase of their evolution once a higher level of divergence is reached. PMID- 11864371 TI - Model-based cluster analysis of microarray gene-expression data. AB - BACKGROUND: Microarray technologies are emerging as a promising tool for genomic studies. The challenge now is how to analyze the resulting large amounts of data. Clustering techniques have been widely applied in analyzing microarray gene expression data. However, normal mixture model-based cluster analysis has not been widely used for such data, although it has a solid probabilistic foundation. Here, we introduce and illustrate its use in detecting differentially expressed genes. In particular, we do not cluster gene-expression patterns but a summary statistic, the t-statistic. RESULTS: The method is applied to a data set containing expression levels of 1,176 genes of rats with and without pneumococcal middle-ear infection. Three clusters were found, two of which contain more than 95% genes with almost no altered gene-expression levels, whereas the third one has 30 genes with more or less differential gene-expression levels. CONCLUSIONS: Our results indicate that model-based clustering of t-statistics (and possibly other summary statistics) can be a useful statistical tool to exploit differential gene expression for microarray data. PMID- 11864372 TI - A database for the provisional identification of species using only genotypes: web-based genome profiling. AB - BACKGROUND: For a long time one could not imagine being able to identify species on the basis of genotype only as there were no technological means to do so. But conventional phenotype-based identification requires much effort and a high level of skill, making it almost impossible to analyze a huge number of organisms, as, for example, in microbe-related biological disciplines. Comparative analysis of 16S rRNA has been changing the situation, however. We report here an approach that will allow rapid and accurate phylogenetic comparison of any unknown strain to all known type strains, enabling tentative assignments of strains to species. The approach is based on two main technologies: genome profiling and Internet based databases. RESULTS: A complete procedure for provisional identification of species using only their genomes is presented, using random polymerase chain reaction, temperature-gradient gel electrophoresis, image processing to generate 'species-identification dots' (spiddos) and data processing. A database website for this purpose was also constructed and operated successfully. The protocol was standardized to make the system reproducible and reliable. The overall methodology thus established has remarkable aspects in that it enables non experts to obtain an initial species identification without a lot of effort and is self-developing; that is, species can be determined more definitively as the database is used more and accumulates more genome profiles. CONCLUSIONS: We have devised a methodology that enables provisional identification of species on the basis of their genotypes only. It is most useful for microbe-related disciplines as they face the most serious difficulties in species identification. PMID- 11864373 TI - The evolutionary history of effectors downstream of Cdc42 and Rac. AB - Cdc42 and Rac--ancient, highly conserved, small GTPases--mediate extracellular signals, triggering changes in transcription and in the actin cytoskeleton. Although dozens of proteins act downstream of these GTPases, a comparison of effector proteins from evolutionarily diverse organisms suggests that six groups of proteins serve as the core machinery for signaling from Cdc42 and Rac. PMID- 11864374 TI - Exploring prokaryotic diversity in the genomic era. AB - Our understanding of prokaryote biology from study of pure cultures and genome sequencing has been limited by a pronounced sampling bias towards four bacterial phyla - Proteobacteria, Firmicutes, Actinobacteria and Bacteroidetes - out of 35 bacterial and 18 archaeal phylum-level lineages. This bias is beginning to be rectified by the use of phylogenetically directed isolation strategies and by directly accessing microbial genomes from environmental samples. PMID- 11864375 TI - Epigenetic reprogramming in the mammalian embryo: struggle of the clones. AB - During preimplantation development in mammals, distinct epigenetic marks on oocyte and sperm DNA are remodeled to an embryonic pattern. A recent study examining global methylation of repetitive elements in various mammals showed that the reprogramming that occurs during normal preimplantation development is aberrant in cloned embryos. PMID- 11864376 TI - Pollen recognition during the self-incompatibility response in plants. AB - Recent work has identified the elusive male (pollen) determinant that underlies self-incompatibility in Brassica (cabbage). The key pollen factor, recognized by the stigma of an incompatible plant, is a small cysteine-rich protein that interacts directly with the receptor domain of a stigma receptor serine-threonine kinase to initiate haplotype-specific pollen recognition and rejection. PMID- 11864377 TI - Structural maintenance of chromosomes (SMC) proteins, a family of conserved ATPases. AB - SUMMARY: The structural maintenance of chromosomes (SMC) proteins are essential for successful chromosome transmission during replication and segregation of the genome in all organisms. SMCs are generally present as single proteins in bacteria, and as at least six distinct proteins in eukaryotes. The proteins range in size from approximately 110 to 170 kDa, and each has five distinct domains: amino- and carboxy-terminal globular domains, which contain sequences characteristic of ATPases, two coiled-coil regions separating the terminal domains and a central flexible hinge. SMC proteins function together with other proteins in a range of chromosomal transactions, including chromosome condensation, sister-chromatid cohesion, recombination, DNA repair and epigenetic silencing of gene expression. Recent studies are beginning to decipher molecular details of how these processes are carried out. PMID- 11864378 TI - [Prevalence of cataract and surgical coverage among adults aged 50 or above in Shunyi District of Beijing, China]. AB - OBJECTIVE: To estimate the prevalence of cataract, surgical coverage rate and the bilateral cataract-blindness burden among adults aged 50 or above in Shunyi District of Beijing, China, for evaluating the cataract status and the efficacy of the prevention blindness program in more than 10 years. METHODS: Cluster sampling was used in randomly selected individuals aged 50 years or older in 28 villages in Shunyi District in fall, 1996. 5 084 individuals received visual acuity test and eye examination, including lens examination by slit lamp. The cataract operation status was also evaluated. RESULTS: The prevalence of cataract was 23.31% among adults aged 50 or above. The prevalence of cataract was increasing by aging. The prevalence of cataract in women and illiterates was significantly higher statistically. The cataract surgical rate had been increasing since 1984. When the bilateral pinhole vision < 3/60 was defined as the blindness criterion, the cataract blindness surgical coverage rate was 56.36%, and the bilateral cataract-blindness burden was 1.63%. When the bilateral presenting vision < 6/60 was defined as the blindness criterion, the cataract blindness surgical coverage rate was 47.79%, and the bilateral cataract-blindness burden was 2.22%. The cataract surgical coverage rates were much lower and the bilateral cataract-blindness burden much higher in women, illiterates and persons aged 70 or above. CONCLUSIONS: The prevalence of cataract is higher and bilateral cataract-blindness burden is heavier in elder individuals. Cataract blindness is still a serious public health problem in elder, women and illiterates after the prevention blindness program having been actively conducted for more than ten years. PMID- 11864379 TI - [Inquiries about methods of primary eye care and prevention of blindness]. AB - OBJECTIVE: To inquire about the methods of developing primary eye care (PEC) and prevention of blindness. METHODS: The methods of developing PEC and prevention of blindness in recent 10 years were analyzed retrospectively. RESULTS: Five grades of PEC network were established. The training courses 1 007 times were held in Nantong, and the epidemiological survey of the blind was carried out for 6 817 259 people. The blind persons were 13 496 in number, and the prevalence of blindness was 0.20%. Cataract operations were performed on 83.16% of the treatable cataract blind persons. The successful rate of cataract surgery was 98.22%. Eight counties became the national advanced counties of prevention of blindness. Nantong city became the first national advanced city of prevention of blindness in 1997. CONCLUSIONS: PEC is the basis of prevention of blindness. To bring PEC into line with the primary health care (PHC) is the key point of developing PEC. Affordable operation, promising quality of the operative procedure and combination of survey and treatment are the three essential factors of developing PEC and prevention of blindness. To create national advanced county and city is the motive force of developing prevention of blindness. PMID- 11864380 TI - [An epidemiological survey of blindness and low vision in Meixian County]. AB - OBJECTIVE: To study the status of blindness and low vision in Meixian County. METHODS: The stratified random sampling rule and the standard grading system of blindness set by WHO were adopted in the study, and 11 327 subjects were enrolled (a sampling fraction of 2.01%) in the whole county. RESULTS: The prevalence of bilateral blindness and low vision was found to be 0.47% and 0.89%; that of monocular blindness and low vision was 0.72% and 0.59%, respectively. Subjects over 50 years old had a significantly higher prevalence of blindness and low vision. The causes leading to blindness and low vision were, in falling order, cataract, glaucoma, ametropia/amblyopia and pterygium. CONCLUSION: The treatment of cataract is the main measure to decrease the prevalence of blindness and low vision. PMID- 11864381 TI - [A pilot study on rapid glaucoma screening]. AB - OBJECTIVE: To screen glaucoma effectively and rapidly and evaluate the specificity and sensitivity of the method. METHODS: 734 subjects were screened in 4 weekend days. All subjects underwent intraocular pressure (IOP) examination with Tonopen, anterior chamber depth measurement with Van Herick method, fundus photography without pupil dilatation and supra-threshold perimetry with Frequency Doubling Perimeter (FDP), Octopus and Humphery automatic perimeter. The cases with positive results in the above examinations should undertake threshold perimetry and fundus stereo photography for the evaluation of the retinal nerve fiber layer (RNFL). RESULTS: Of the 734 subjects with complete data, there were 55 cases with open angle glaucoma (7.5%), 59 angle-closure glaucoma (8.4%) and 98 glaucoma suspects (13.4%). The IOP of 34 patients was > 21 mm Hg in all the glaucomatous patients (29.8%). The abnormal rates of optic nerve and visual field were 94.5% and 67.3% respectively in the patients with open-angle glaucoma. The abnormal rates of anterior chamber, visual field and optic nerve were 89.8%, 72.9% and 45.8% respectively. CONCLUSIONS: The most efficient method for screening open-angle glaucoma is fundus stereo photography with evaluation by an expert. Anterior chamber evaluation is necessary in screening glaucoma. Supra threshold perimetry has limited specificity, and FDP might be a promising method in screening glaucoma with relatively good specificity. PMID- 11864382 TI - [Prevalence survey on pterygium in two counties of Hainan Province]. AB - OBJECTIVE: To explore the relationship between pterygium and environmental condition, such as area and weather, and human factors, such as sex and race, through the prevalence survey of pterygium in Hainan Province. METHODS: As a sample, all villagers in 14 villages were chosen by random sampling from all the rural villages of Dongfang and Changjiang Counties, the pterygium was examined and their pterygia were recorded and graded according to the shape and the location of the head of pterygium. RESULTS: In 7 990 villagers examined, there were 628 cases with pterygium, prevalence being 7.86% and including 439 cases (8.28%) in 5 303 Han people and 189 cases (7.03%) in 2 687 Li people. The difference of the prevalence between Han and Li nationality was significant (chi(2) = 3.897, P = 0.048). There were 266 male cases (6.43%) and 362 female cases (9.40%). The prevalence in male was less than that in female (chi(2) = 19.852, P = 0.001). The distribution of the pterygium in eyes was symmetrical. CONCLUSION: The prevalence of pterygium in Hainan is higher than that in other provinces of China. It may be related to the geographic location and weather condition. In Hainan Province, the prevalence of pterygium in Han nationality is higher than that in Li nationality and in female higher than that in male. The increase of the prevalence of pterygium may be related to the increase of age. PMID- 11864383 TI - [A research of infant refraction in Kunming Municipality]. AB - OBJECTIVE: To study the infant refraction, detect ametropia in mass screening refraction and correct the ametropia properly for the treatment and prevention of amblyopia and strabismus as early as possible. METHODS: The refraction status of 1 146 children (2 291 eyes, aged 1 month -3 years) were determined with retinoscopy after tropicamide cycloplegia. RESULTS: The results of statistics showed that 89 eyes (3.88%) had emmetropia, 2 139 eyes (93.37%) hypermetropia, 38 eyes (1.66%) myopia, 560 eyes (24.44%) astigmatism and 25 eyes (1.09%) mixed astigmatism. In cases with ametropia, most of them were mild, and 97 eyes (4.23%) were moderate and severe. Anisometropia occurred in 34 infants (2.97%), and 7 infants (11 eyes, 0.48%) had strabismus. The degree of hypermetropia decreased with the increase of age. The percentage of astigmatism decreased with the age increase (chi(2) = 7.46, P<0.01), and the degree of astigmatism also decreased with the age increase (chi(2) = 26.1l, P < 0.01). Myopia increased with the age increase (chi(2) = 4.06, P < 0.05). CONCLUSIONS: The prevention of amblyopia and strabismus in children should begin at the infant period, and the cases with moderate and severe ametropia should wear eye glasses as early as possible. PMID- 11864384 TI - [Patient awareness study of elderly population in rural Guangdong]. AB - OBJECTIVE: To investigate the patients' awareness on the eye diseases, treatment and major barriers for patients to seek eye care. METHOD: Randomly clustering sampling method was used for all the subjects aged 50 years and over, and their eyes were examined. All subjects with presenting visual acuity less than 0.1 in at least one eye underwent the test of the patient's awareness questionnaire developed in Aravind Eye Hospital, India. RESULTS: Five hundred and thirty-five (85.5%) visual impairment subjects were successfully interviewed. In 526 cases with the ocular disease course over 1 year, 95% of them were aware of the occurrence of eye diseases for more than 1 year. However, in the 448 cases with the ocular diseases treatable, only 24.2% of them realized that their eye diseases could be treated, while 73% of them did not know or did know right on the examination date. The major barriers for seeking eye care were economic reason, fear of operation, far distance from a hospital, etc. CONCLUSION: Patients' education and low price service are probably the effective methods to clear the roadblock on the way of improving the coverage of eye care. PMID- 11864385 TI - [A clinical study on black diaphragm posterior chamber intraocular lens implantation]. AB - OBJECTIVE: To discuss the effect of black-diaphragm intraocular lens implantation in the management of cataract or aphakia with aniridia. METHODS: Thirty-three cases (34 eyes) of black-diaphragm posterior chamber intraocular lens implantation were retrospectively analyzed, among which 18 eyes had undergone primary intraocular lens implantation and 16 secondary intraocular lens implantation. RESULTS: After 1-52 months of follow-up, postoperative corrected visual acuities were greater-than-or-equal 1.0 in 17 eyes, 0.5-0.9 in 11 eyes and 0.1 -0.4 in 6 eyes. The complications included intraoperative ciliary body bleeding in 2 eyes and retinal detachment in one eye. CONCLUSION: Black-diaphragm intraocular lens implantation is effective in the management of cataract or aphakia with aniridia. PMID- 11864386 TI - [Clinical observation of phacoemulsification with foldable intraocular lens implantation through a limbus incision]. AB - OBJECTIVE: To investigate the clinical therapeutic effects of phaco emulsification with foldable intraocular lens implantation through a limbus tunnel incision. METHODS: Eighty-six patients (107 eyes) with senile cataract underwent phacoemulsification with acrylic foldable posterior chamber intraocular lens implantation through a 4 mm superior limbus tunnel incision. RESULTS: The corrected visual acuity of 0.5 or better was 62.6% at the 1st day, and 76.6%, 92.5% and 88.8% respectively at the 1st week, 1st month and 3rd month postoperatively. CONCLUSION: Phacoemulsification with foldable intraocular lens implantation through a limbus tunnel incision has advantages of both scleral tunnel incision and clear corneal tunnel incision. The postoperative therapeutic effects are satisfactory. PMID- 11864387 TI - [Revision of thin-walled cystic bleb by transposing conjunctival flap technique]. AB - OBJECTIVE: To investigate the clinical effect of transposing conjunctival flap technique on thin-walled cystic blebs after trabeculectomy for treatment of glaucoma. METHODS: Fifty-two eyes of 46 patients with thin-walled cytic blebs were underwent this modified operation. The procedure involved incising the conjunctiva surrounding the preserved bleb, then the conjunctiva-Tenon's fascia flap superior or temporal to the cystic bleb was relaxed and mobilized to cover the bleb surface after topical cryopexy. The anterior margin of the flap was sutured onto the peripheral cornea. In order to relax the conjunctival flap sufficiently, an incision was made close to the superior fornix above the flap and parallel to limbus to decrease the tension. RESULTS: In the follow-up of 6 months-4 years, the mean intraocular pressure (IOP) of 52 eyes increased to postoperative (12.98 plus minus 5.70) mm Hg from preoperative (4.25 plus minus 1.33) mm Hg (1 mm Hg = 0.133 kPa) (t = 3.26, P < 0.01), 51 eyes maintained the filtering function. Post-operatively, the bleb leakage in 20 eyes and macular edema in 6 eyes disappeared. Local antiglaucoma medication was required in one eye, because the bleb disappeared with increasing IOP. The post-operative visual acuities were elevated in comparison with that before the operation (chi(2) = 16.65, P < 0.01). CONCLUSION: The transposing conjunctival flap technique is an effective surgery for the repair of thin-walled cystic bleb whose complications, such as hypotony, maculopathy, bleb leakage or endophthalmitis can be cured or prevented. PMID- 11864388 TI - [MRI diagnosis of painful ophthalmoplegia syndrome]. AB - OBJECTIVE: To study MRI appearances of painful ophthalmoplegia syndrome and provide image basis for clinical diagnosis of painful ophthalmoplegia syndrome. METHODS: Seventeen cases with painful ophthalmoplegia syndrome diagnosed clinically were collected. MRI ordinary scan and gadolinium-diethylene triamine pentaacetic acid (Gd-DTPA) contrast fatty-restraint scan were performed on the orbit and cavernous sinus. RESULTS: MRI appearances of painful ophthalmoplegia syndrome included patch or small nodule abnormal signal at the region of fissura orbitalis superior. The ipsilateral cavernous sinus was enlarged. The local lobus temporalis meninges close to the fissura orbitalis superior and cavernous sinus were invaded. MRI contrast fatty-restraint scan might clearly show the pathological changes. CONCLUSION: MRI may provide image basis for the clinical diagnosis of painful ophthalmoplegia syndrome. PMID- 11864389 TI - [Influence on tear film of postoperative 5-fluourouracil and intraoperative mitomycin C in glaucoma filtration surgery]. AB - OBJECTIVE: To investigate the changes of tear film functions due to the use of postoperative 5-fluourouracil (5-Fu) and intraoperative mitomycin C (MMC) in glaucoma filtration surgery. METHODS: 71 eyes of 71 refractory glaucoma patients were followed up after filtration surgery. They were divided into 3 groups. The 5 Fu and MMC groups consisted of 24 and 17 eyes respectively. The other 30 eyes that underwent the surgery without any use of antimetabolic agents were in the control group. The observed symptoms included ocular foreign body sensation, dryness, photophobia, itching and discharge. The available criteria were conjunctival congestion, fluorescein and rose bengal staining on the cornea and conjunctiva, break-up time of tear film and Schirmer's test. RESULTS: The ocular symptoms and signs were manifest and tear film functions were significantly decreased in 5-Fu group (P < 0.05). No significant difference between MMC and the control was seen, except that the dry feeling was significantly intensive in MMC group (P < 0.05). CONCLUSION: The postoperative use of 5-Fu after glaucoma filtration surgery leads to tear film abnormalities apparently. PMID- 11864390 TI - [The influence of flexible open-loop anterior chamber intraocular lens on the structure of ocular anterior segment]. AB - OBJECTIVE: To evaluate the therapeutic effects and safety of flexible open-loop anterior chamber intraocular lens (FOAC-IOL). METHODS: By ultrasound biomicroscopy, the positions of the haptics of FOAC-IOL in 20 patients (20 eyes) and the relationships of the haptics with iris and anterior chamber angle structures were observed postoperatively to evaluate the impact of these lenses on the structures of ocular anterior segment. Follow-up duration was from 6 to 20 months. RESULTS: Among 40 IOL's haptics, twenty-six haptics were fixed at the recess of the chamber angle; and other fourteen haptics penetrated the iris and invaded into the ciliary stroma, which caused recurrent uveitis. There was 1 eye with eccentric lens. There were 8 eyes with anterior synechiae of iris, which were related to the haptics of intraocular lens. CONCLUSIONS: Ultrasound biomicroscopy works well in dynamic observation of the influence of intraocular lens on ocular anterior segment. The haptics of flexible open-loop anterior chamber intraocular lens should be modified. PMID- 11864391 TI - [Dexamethasone inhibits trabecular meshwork cell growth and epidermal growth factor mRNA expression in vitro]. AB - OBJECTIVE: To investigate the effect of dexamethasone on cultured trabecular meshwork cell growth and the cell expression of epidermal growth factor (EGF) mRNA in vitro. METHODS: Human trabecular meshwork cells were cultured in vitro. The third passage cells were used in this experiment; with the addition of 300 microgram/ml dexamethasone in the culture medium in the test group, the culture was carried out as routine. After 5 days, the cell growth conditions were observed. After 7 days, the cells in the test and the control group were collected, and mRNA was drawn from them in both groups. EGFcDNA probe with alpha (32)P isotope labeling was used to proceed dot blot hybridization and autoradiography to detect EGFmRNA of the cells. A computerized laser instrument was used to scan the size of dot blot hybridization, the relative value of the optical density was measured in the autoradiographs, and the inter-group comparison was made. RESULTS: Trabecular meshwork cells were inhibited by dexamethasone in the test group. The cells in the control group had been confluent at the 5th day, but the cells in the test group still grew like tribes. The total RNA was 14 microgram in the test group, while 22.5 microgram in the control group. In both groups, 14 microgram RNA was used for dot blot hybridization test with EGFcDNA probe. Autoradiographic image was positive in the two groups. The results of the scanning and the measurement of the autoradiograph density showed that dexamethasone obviously decreased the EGFmRNA expression of trabecular meshwork cells. CONCLUSIONS: Dexamethasone can inhibit trabecular meshwork cell growth in vitro. At first it affects the cell total RNA transcription and the second affects the EGFmRMA expression. It is suggested that steroid glaucoma be caused by the inhibition of various physiological and metabolic functions of trabecular meshwork cells induced by dexamethasone. PMID- 11864392 TI - [T cell and molecule-1 detection in cornea of fetus, neonates, children and adults]. AB - OBJECTIVE: To study the immune state of human cornea with different ages. METHODS: The human corneas in different ages were detected by using immunohistochemical techniques for 5 types of monoclonal antibody. RESULTS: The contents of T-cell (CD(+)(3)), helper/inducer (CD(+)(4)), toxic T-cell (CD(+)(8)), macrophage (CD(+)(68)) and molecule-1 (intercellular adhesive molecule, CD(+)(54)) in the adult cornea were relatively low, mainly in the limbus but none at the corneal center. These positive cells were less in corneas of fetus, neonates and children than that in adults. CONCLUSION: In corneas of fetus, neonates and children, the T-cell, macrophage and molecule-1 were less than that of adults. Their corneas may be used as donors in corneal transplantation. PMID- 11864393 TI - [Ultrastructural change of rabbit lens epithelial cells induced by low power level microwave radiation]. AB - OBJECTIVE: To study the ultrastructural change of rabbit lens epithelial cells with transmission electron microscope and to observe whether low power level microwave can induce the injury of rabbit lens. METHODS: Eight rabbit eyes were exposed to 10 mW/cm(2) low power level microwave radiation for 30 hours. Transmission electron microscope was used to observe the ultrastructural changes of the lens epithelial cells. The other eye was served as self-control. RESULTS: The ultrastructure of exposed lens epithelial cells changed apparently. The characteristic apoptosis cell morphological changes, such as small cell size, the decreased number of mitochondria, balloon-shaped mitochondria, etc can be seen. No obvious changes can be found in the protected self-control eyes. CONCLUSIONS: The low power level microwave (10 mW/cm(2)) can induce the ultrastructural changes of rabbit lens epithelial cells, this may be the appearance of early irreversible microwave radiation injury. In regard to the pathogenic mechanism, it is necessary to perform further investigations. PMID- 11864394 TI - [Apoptosis of retinal capillary cells in diabetic rats and expression of apoptosis-related genes]. AB - OBJECTIVE: To determine the apoptosis of retinal capillary cells in early diabetic rats and observe the expression of apoptosis-related genes (bcl-2, Bax) in retinas of early diabetic rats. METHODS: 40 healthy adult Wistar rats were chosen and randomly divided into four groups: normal control (CON), diabetes mellitus one month (DM1), three months (DM3) and six months (DM6). Diabetic mellitus in rats was induced by streptozotocin (STZ) intraperitoneally. Retinal preparations of blood vessels and paraffin sections were stained by TdT-mediated dUTP nick end labeling (TUNEL) and immunohistochemical (ABC) method respectively and analyzed by computer-picture analytic system on the results of ABC method. RESULTS: Positive pericyte nuclei labeled by TUNEL were emerged in DM3 and DM6, but positive endothelial cell nuclei only in DM6. No positive reaction was seen in CON and DM1. Chromatin of TUNEL positive nuclei was distributed unequally, showing apoptotic characteristics of ring-like and crescent nuclei. In ABC staining method, two genes' (bcl-2, Bax) proteins were expressed in retinal vessels. Along with the progression of diabetes, the intensity of positive reaction was increased in degrees. Moreover, pigment epithelial cells in DM3 showed Bcl-2 positive reaction that again extended further to inner rod segments and ganglion cells in DM6, while in DM6, Bax was expressed in ganglion cells. CONCLUSIONS: The nature of pericyte dropout in early diabetic rats is apoptosis that also exists in endothelial cells; the expression of Bax and bcl-2 in retinas of early diabetic rats is enhancing. PMID- 11864395 TI - [Survey on the growth of interpupillary distance of Chinese children aged 5 to 17 years]. AB - OBJECTIVE: To investigate the interpupillary distance (IPD) of Chinese children for stipulating the sizes for children spectacle frames. METHOD: The IPD of 10171 children aged 5 to 17 years old was measured with a caliper in four cities. RESULTS: The IPD was positively correlated with the age increase from 5 to 17 years. The IPD in males was larger than that in females. The differences of IPD between males and females in the age groups of 5 to 9 and 10 to 15 years were statistically significant (F = 400.97, P < 0.01). CONCLUSIONS: There are two peak periods of the growth of IPD in Chinese children at 5 to 9 and 10 to 15 years. The IPD in males reaches adult level at age 15 and in females at age 13. The differences of the IPD between male and female and among 4 cities have no practical meaning in stipulating the sizes of children spectacle frames. PMID- 11864396 TI - [Immunohistochemical and clinical studies on sub-corneal flap epithelial implantation accompanied by flap melting after excimer laser in situ keratomileusis]. AB - OBJECTIVE: To study the immunohistochemical and clinical characteristics of the corneal epithelial cells implanted under the flap accompanied by the flap melting after excimer laser in situ keratomileusis (LASIK) and the method of management. METHODS: In eight eyes (In 4 eyes a part of the flap melted.) with corneal epithelial cells implanted under the flap after LASIK, the corneal flap was torn again. The implanted tissue was removed. After washing between the interfaces, the flap was repositioned. The implanted tissue was immunohistochemically stained by using four kinds of monoclonal antibodies and using terminal deoxyribonucleotide transferase-mediated dUTP nick end labeling (TUNEL) method. RESULTS: Early in the course, the disorder appeared on the temporal side, expanded and thickened. The defective periphery of the corneal flap presented as a partial lunar eclipse. The melting process was self-limited in 3 months. The result was good after the implanted tissue was removed. The implanted material was stained by using HE, immunohistochemical and TUNEL methods. Some vesicle-like degeneration in the implanted layer was found. There was lysozyme in the vesicles, and the cells of implanted layer possessed a trend of apoptosis. All the cells were dead in the vesicles. CONCLUSIONS: The progress of corneal epithelial implantation under corneal flap after LASIK may include 3 periods: epithelial implantation, corneal flap melting and self-limitation. Early in the course, the implanted material presented as a thin film. Afterwards, it becomes a non-structural material. Possibly the melting of the corneal flap is caused by apoptosis. Therefore, the implanted epithelium should be removed early. PMID- 11864397 TI - [Studies on the change of extracellular matrix in the corneal tissue after photorefractive keratectomy in rabbits]. AB - OBJECTIVE: To investigate the relationship between the haze formation and the change of extracellular matrix (ECM) in the corneal tissue after photorefractive keratectomy (PRK). METHODS: Twenty-four adult New Zealand White rabbits received PRK in the right eye for minus sign10.00 D ablation. According to the time of rabbits sacrifice, they were divided into 8 groups, and 3 rabbits were in each group. They were immediate, 24 hour, 1 week, 2 week, 1 month, 3 month, 6 month and 12 month groups. After surgery, the corneas were examined periodically by slip-lamp microscopy for haze formation. The right eyes were enucleated, and the corneas were studied with immunohistochemical evaluation for extracellular matrix, including collagen type I, type III, type VI, cellular fibronectin, tenascin and laminin. RESULTS: The incidence of haze was 100% after PRK. The formation of haze had a tendency from mild to severe, and along with the time prolongation, it gradually decreased and disappeared. The newly synthesized collagen type I, type III, type VI, cellular fibronectin, tenascin and laminin were deposited in the anterior corneal stroma. CONCLUSION: The formation of haze is closely related to the deposition of ECM elements in the anterior corneal stroma in the surgical area. PMID- 11864398 TI - [Changes in Descemet's membrane and endothelium after photorefractive keratectomy in rabbits]. AB - OBJECTIVE: To investigate the effects of epithelial abrasion on the corneal endothelium and Descemet's membrane and to compare the changes with those after eximer laser photoablation of the cornea. METHODS: The animal models of corneal epithelial abrasion and excimer laser photoablation of the cornea were performed on the rabbit eyes. The central epithelial abrasion, 7 mm in diameter, was created by mechanical removal of the epithelium, and the specimens were examined at 24 hours, 7 days, 14 days and 30 days by transmission and scanning electron microscopy. Corneas that were photoablated by an excimer laser and non-treated normal corneas were investigated as controls. RESULTS: (1) Corneas with the epithelium removal showed massive enlargement of the mitochondria in the endothelium and exhibited a layer of electron-dense fibrillogranular material that had migrated forward through the Descemet's membrane. These alterations were similar to the changes observed after photoablation of the cornea by an excimer laser. (2) The edema and pathologic changes of corneal endothelium in eyes with denuded epithelium alone were similar to that observed after photoablation of the cornea by an excimer laser. CONCLUSION: The extrusion of electron-dense material in the Descemet's membrane observed after excimer laser ablation destruction of the epithelial integrity. PMID- 11864399 TI - [Excimer laser in situ keratomileusis for severe ametropia after penetrating keratoplasty]. AB - OBJECTIVE: To evaluate the effects of excimer laser in situ keratomileusis in correcting severe myopia and astigmatism after penetrating keratoplasty. METHODS: Excimer laser in situ keratomileusis was performed on ten eyes of ten patients to correct high ametropia in cases having previously undergone penetrating keratoplasty. RESULTS: After surgery, all grafts remained clear and no corneal graft rejection occurred during the follow-up period. The average spherical equivalent refraction decreased from minus sign11.85 D preoperatively to minus sign3.38 D postoperatively and the average cylinder from 6.75 D to 2.70 D. The highest cylinder correction achieved was 10.00 D. The mean uncorrected visual acuity improved from 0.1 to 0.8, the best one was 1.5, and the best corrected visual acuity (BCVA) improved from 0.8 to 1.1. In this group, BCVA was improved in all eyes. CONCLUSION: Excimer laser in situ keratomileusis is a safe and effective method to deal with the high ametropia after penetrating keratoplasty and shows its great prospect. PMID- 11864401 TI - [Change of cone's motion perception in patients with primary open angle glaucoma]. AB - OBJECTIVE: To assess the change of cone's motion perception in primary open angle glaucoma (POAG). METHODS: Equi-luminance random dots moving test was used to determine the change of motion perception of blue, green and red cones in 51 eyes with POAG. To isolate the three kinds of cones, the blue, green and red visual targets were displayed on the yellow, purple and blue-green background respectively. RESULTS: The direction discrimination thresholds of motion perception from blue, green and red cones were all damaged in POAG compared to that of the normal group, and among them the blue cone was the most obviously damaged. CONCLUSIONS: These findings indicate that cone's motion perception, especially the blue cone's, may reveal pre-clinical visual nerve damage in POAG. PMID- 11864400 TI - [The characteristics of multifocal electroretinogram in normal subjects in China]. AB - OBJECTIVE: To test the characteristics of multifocal ERG in normal Chinese subjects and to provide reference values in clinical application. METHODS: The multifocal ERGs by using VERIS 4.0 visual evoked response imaging system were tested for 42 subjects (48 eyes). Their ages ranged from 11 to 67 years (mean, 40.7 years). The stimulative visual angles subtended plus minus 26.6 horizontally and plus minus 22.1 vertically. Burian-Allen contact lens electrode was used to record the responses from 103 retinal locations in 8 min (16 segments). RESULTS: With age increasing, the latencies of N(1) wave and P(1) wave delayed, there were statistically significant differences of the latencies between < 50 year group and greater-than-or-equal 50 year group (t > 2.01, P < 0.05), and the amplitude densifies of N(1) wave and P(1) wave decreased with wide inter-individual variation. The amplitude densities of N(1) wave and P(1) wave were greatest at the central fovea, they were (44.88 plus minus 13.30) nV/deg(2), (70.90 plus minus 17.77) nV/deg(2) respectively, the densities reducing towards more peripheral location. The amplitude sum of N(1) wave at the upper retina were greater than that at the lower retina, and the amplitude densities and sums of N(1) wave and P(1) wave at superior-temporal quadrant were greater than that at inferior-nasal quadrant. CONCLUSION: Our results suggest multifocal ERG be able to reflect the visual function at different retinal locations correctly and objectively and provide normal reference limits. PMID- 11864402 TI - [Clinical observation of astigmatism induced by corneal incision after phacoemulsification]. AB - OBJECTIVE: To evaluate the changes of astigmatism induced by corneal incision after phacoemulsification. METHODS: Phacoemulsification was performed on 62 cases (78 eyes) without suture for a corneal incision. The changes of corneal astigmatism before and after operation in group A (incision at the superior nasal or superior temporal) and B (incision at the steepest corneal meridian) were compared. RESULTS: Three months after the operation, the surgical induced astigmatism was (0.83 plus minus 0.65) D and (0.72 plus minus 0.55) D in group A and B respectively. There was no significant difference, statistically (P > 0.05). The changes of corneal astigmatism were 0.11 D and 0.39 D in group A and B respectively, the difference being statistically significant (P < 0.05). CONCLUSION: Surgically induced astigmatism is very small after a corneal incision in phacoemulsification without a suture. If the incision is placed on the steepest meridian, the corneal astigmatism can be significantly reduced postoperatively. PMID- 11864403 TI - [Pediatric phacoemulsification with Acrysof intraocular lens implantation]. AB - OBJECTIVE: To investigate the operative effects of pediatric phacoemulsification with Acrysof intraocular lens (IOL) implantation. METHODS: Among 153 eyes of 120 children with cataract aged 3 -- 12 years old, phacoemulsification with Acrysof IOL implantation was performed on 76 eyes of 61 children, and cataract extracapsular extraction with polymethylmethacrylate (PMMA) IOL implantation was performed on 77 eyes of 59 children. The visual acuity, percentages of various complications and rate of neodymium: YAG (Nd: YAG) capsulotomy were followed up for 6 -- 18 months. RESULTS: There were significant differences in the percentages of various complications and rate of neodymium: YAG (Nd: YAG) capsulotomy between the Acrysof group and the PMMA group (P < 0.01). The Acrysof group was associated with less posterior capsular opacification (6.6%) than the PMMA group (80.5%). The YAG capsulotomy rate was 2.6% for the Acrysof group, and 64.9% for the PMMA group. After operation 100% and 90.6% patients were relieved from blindness in the Acrysof group and the PMMA group respectively, the difference being not statistically significant (P > 0.05). 72.6% patients in the Acrysof group and 51.5% in the PMMA group obtained corrected visual acuity greater-than-or-equal 0.3, and the difference was statistically significant (P < 0.05). CONCLUSIONS: Phacoemulsification with Acrysof intraocular lens implantation is effective in pediatric cataract surgery. Most of the patients obtain remarkable improvement in visual acuity with minimal complications and YAG capsulotomy rate after operation. It is necessary to perform further randomized and prospective studies to evaluate the long-term postoperative effects of Acrysof IOLs used in pediatric cataract surgery. PMID- 11864404 TI - [Reversed trabeculectomy for treatment of glaucoma]. AB - OBJECTIVE: To reduce the postoperative blocking of the filtration canal in routine trabeculectomy with modification. METHODS: The modification of the procedure of routine trabeculectomy was that one end of the 4.0 mm x 1.5 mm deep layer scleral tissue including part of trabecular meshwork was not cut. Instead, the free end of the strip of deep scleral tissue was vertically rotated and sutured on the deep layer of the scleral bed posterior to it. The projection of the strip was utilized to prevent postoperative blocking of filtration canal. RESULTS: The mean period of postoperative follow-up was 23.5 months (range, 12 -- 36 months). In the treatment group (38 eyes of 33 cases), the intraocular pressure (IOP) was lowered from preoperative (25.41 plus minus 2.41) mm Hg to postoperative (14.74 plus minus 0.90) mm Hg. The IOP was lowered from preoperative (24.96 plus minus 1.35) mm Hg to postoperative (19.40 plus minus 1.43) mm Hg in the control group (40 eyes of 33 cases). The success rates were 94.7% and 77.5% in the treatment and control group respectively. CONCLUSIONS: The modified trabeculectomy is safe, easy to perform, and can effectively relieve the postoperative blocking of the filtration canal, control postoperative IOP and reduce postoperative recurrence. PMID- 11864405 TI - [Penetrating keratoplasty in active herpes simplex stromal keratitis]. AB - OBJECTIVE: To evaluate the efficacy of penetrating keratoplasty (PKP) in treating active herpes simplex stromal keratitis. METHODS: PKP was performed on 59 patients with quiescent and 66 patients with active herpes simplex stromal keratitis. The results and complications of herpes simplex keratitis (HSK) in the quiescent group were compared with that of the HSK active group. RESULTS: No clinical or statistical significant differences in visual outcome, graft transparent rate and recurrent rate of HSK after operation were identified between the active and quiescent groups. CONCLUSION: When the pre- and intra operative managements of PKP are properly performed on active HSK, the same results as that performed on quiescent HSK can be obtained, and it can reduce the complications and shorten the duration of the disease. PMID- 11864406 TI - [The comparison of two methods to predict the post-operative visual acuity of cataractous patients]. AB - OBJECTIVE: To study the accurate methods to predict the post-operative visual acuity of cataractous patients with different degrees of opaque lenses. METHODS: Visual electrophysiological methods and retinometer were used to evaluate the visual function after phacoemulsification and intraocular lens implantation in 152 cases (152 eyes). The regressive analyses between the post-operative best corrected visual acuity (BCVA) and each of the following results: the pre operative flash electroretinogram (F-ERG), flash visual evoked potentials (F-VEP) and potential visual acuity (PVA) were performed, respectively. RESULTS: More obvious linear-relationship was shown between PVA and BCVA (r = 0.654) than that between the combination of b wave's amplitude of F-ERG with wave's amplitude of F VEP and BCVA (r = 0.528) in the patients with partial opaque lenses. Obvious linear-relationship between the combination of the F-ERG with F-VEP values and BCVA (r = 0.487) and no obvious relationship between the potential visual acuity and BCVA (r = 0.049) were shown in the patients with near complete/complete opaque lenses. CONCLUSION: Both visual electrophysiological and retinometric detection, especially the latter one, can predict the post-operative visual acuity of the patients with partial opaque lenses accurately. Visual electrophysiological detection can predict the post-operative visual acuity of cataractous patients with near complete/complete opaque lenses accurately but retinometric detection can not. PMID- 11864407 TI - [Corneal topography and pachymetry in normal eyes]. AB - OBJECTIVE: To evaluate the corneal topography and pachymetry on normal eyes. METHODS: The Orbscan corneal topography system was used to investigate corneal topography and pachymetry on 46 normal subjects. RESULTS: In normal eyes, island shape was the most common type in the elevation topography of anterior corneal surface, followed by incomplete ridge, regular ridge, irregular ridge and unclassified type. For the elevation topography of posterior corneal surface, the most common type was also the island-shape, followed by regular ridge, incomplete ridge and irregular ridge. The thinnest point of cornea in normal eyes was located (0.90 plus minus 0.51) mm away from the visual axis with the thickness of (0.55 plus minus 0.33) mm, which was commonly seen at the inferotemporal quarter of cornea, followed by superotemporal, inferonasal and superonasal quarter. The mean corneal thickness was the least in corneal center and greatest in superior part of cornea. For the color-coded map of corneal thickness, oval shape was the commonest type, followed by round shape, eccentric oval shape and eccentric round shape. CONCLUSIONS: The normal parameters on pan-corneal thickness and corneal shape of both anterior and posterior surface will provide much more useful informations in finding abnormal cornea. Orbscan topography system is a useful instrument for the evaluation of corneal shape and thickness, which is helpful in corneal refractive surgeries. PMID- 11864408 TI - [A study of different intracellular signal transduction pathways on phagocytosis of fibronectin by retinal pigment epithelial cells]. AB - OBJECTIVE: To determine the phagocytosis of fibronectin (FN) by retinal pigment epithelial (RPE) cells and its related intracellular signal transduction pathways. METHODS: Fluorescent latex beads were coated by FN and then incubated with RPE cells at 37C for 3 hours. Phagocytosis was quantified by a flow cytometric assay. Experiments were also performed in the presence of inhibitors of various intracellular signaling pathways [tyrosine kinase (TK), phosphatidylinositol 3-kinase (PI 3-K), protein kinase C (PKC)]. RESULTS: FN coated beads produced significantly increase in the phagocytic index (P < 0.05) when compared to the uncoated control. The PKC inhibitors, phorbol 12-myristate 13-acetate (PMA, 100 nmol/L) or calphostin C (400 nmol/L), non-specifically increased the phagocytosis of both FN-coated (P < 0.01) and uncoated beads (P < 0.01). Inhibitors of TK (genistein, 100 microgram/ml) and PI 3-K (wortmannin, 5 micromol/L), significantly inhibited FN phagocytosis (P < 0.001) but did not affect the uncoated control. While a combination of different inhibitors (genistein plus wortmannin, genistein plus PMA, wortmannin plus PMA) was used, the results showed that genistein or wortmannin can counteract the effect of PMA, and that genistein plus wortmannin have an additive inhibitory effect. CONCLUSIONS: Our results suggest that the FN phagocytosis by RPE cells appear to be regulated, at least in part, by some signal transduction pathways. The knowledge of the signaling pathways that mediate FN phagocytosis by RPE cells may provide novel therapeutic targets for molecular pharmacology of proliferative RPE disorders such as proliferative vitreoretinopathy. PMID- 11864409 TI - [An immunohistochemistry study on extracellular matrix of lamina cribrosa in optic nerve head of experimental glaucoma]. AB - OBJECTIVE: To observe the change of the macromolecular components of extracellular matrix of lamina cribrosa (LC) in experimental glaucoma. METHODS: Immunoperoxidase (ABC) staining method was used to investigate the changes of collagen type IV and laminin in the lamina cribrosa of 9 monkey eyes with experimental glaucoma. RESULTS: At the late stage of glaucoma, the labeling of collagen type IV and laminin were increased at pre-LC and LC area due to the accumulation of basement membrane-like materials. CONCLUSION: The changes may be a selective response to elevated intraocular pressure. This response may alter the biochemical composition and the biomechanics of the lamina cribrosa. PMID- 11864410 TI - [The effect of viper venom on the attachment, migration and proliferation of Tenon's capsular fibroblasts]. AB - OBJECTIVES: To observe the effect of viper (Ahylysantipfarciasi) venom on attachment to collagen, migration and proliferation of rabbit Tenon's capsular fibroblast (TFs) in tissue culture. METHODS: Anti-adhesion experiment: The 3 -- 5 passage TFs suspended in DMEM medium were pre-incubated for 30 minutes in the presence of various concentrations of viper venom (0, 2.5 x 10(-4), 5.0 x 10(-4), 1.0 x 10(-3) and 5.0 x 10(-3) U/ml) at 37C, 5% CO(2), then inoculated in 24-well plate coated with rat-tail collagen. After incubation for 90 minutes, the floating cells were removed. The attached cells in each well were enumerated microscopically and determined by the value of absorption (A) of 3-(4, 5 dimethylthiazolzyl)-2, 5-diphenyl tetrazodium bromide (MTT). Anti-migration experiment: A denuded area was made when the 3rd passage of TFs was confluent into a monolayer. Cells were then exposed to viper venom (0 -- 5.0 x 10(-3) U/ml) at 37C, 5% CO(2), and the cells having migrated into the denuded area were enumerated every 6 hours. Anti-proliferative experiment: Two hours after the 3rd passage of TFs was incubated in the 24-well plate at 37C, 5% CO(2), they were exposed to different concentrations of viper venom drug (0 -- 5.0 x 10(-3) U/ml). Twenty-four and 48 hours later, the number of cells was determined by MTT method. RESULTS: Viper venom inhibited TFs from attaching to collagen in a dose-dependent manner and the ID(50) was 1.0 x 10(-3) U/ml. Only did 5.0 x 10(-3) U/ml of viper venom show significant difference from the control during 6 -- 48 hours. The difference of A value was not significant among all groups at 24 and 48 hr. CONCLUSIONS: In vitro, viper venom (> 1.0 x 10(-3) U/ml) can significantly inhibit the attachment of TFs to collagen, and 5.0 x 10(-3) U/ml can inhibit the migration, but can not affect their proliferation. PMID- 11864411 TI - [Efficacy of topical cyclosporine A on keratoplasty rejection in rats]. AB - OBJECTIVE: To evaluate the efficacy of topical cyclosporine A (CsA) on a penetrating keratoplasty rejection model in rats. METHODS: Lou rats received orthotopic corneal allografts from inbred F344 donors. The rats were treated with one drop of topical solution four times daily for 30 days. Sixty rats were divided into one control (the matrix of CsA solution) and five treatment groups (0.5%, 1.0%, 2.0% CsA, glucocorticoid, glucocorticoid and 1.0% CsA). Rejection index (RI) and mean survival time (MST) were calculated, and histopathological and immunohistopathological examination were performed for each group. RESULTS: The graft MST was postponed by topical CsA treatment. There was significant difference between each treatment group and the control group (t greater-than-or equal 2.28, P < 0.0l), especially 2% CsA, steroid and combined 1% CsA and steroid groups, the combined group being the most effective. The RI was lower in all the groups with topical CsA than that in the control group (P < 0.01). The effect of 2% CsA was the same as that of the steroid, while 1% CsA combined steroid was the most effective. The histopathological findings confirmed that the topical application of CsA reduced the inflammatory cells infiltrating the graft stroma and the neovascularization. Immunohistopathological analysis demonstrated that there were fewer CD(+)(11a) T cells and macrophages in the infiltration and intercellular adhesion molecule (ICAM)-1 positive cells in keratocytes and endothelium of grafts in the treatment groups than in the control group. CONCLUSIONS: Topical CsA can inhibit the keratoplasty rejection episodes in the rat and afford better results when combined with topical steroid. PMID- 11864412 TI - [Vitreous surgery for proliferative maculopathy]. AB - OBJECTIVE: To explore the indication of vitrectomy for the treatment of pre macular proliferative disease, the surgical technique, results and the prevention and treatment of complications. METHODS: Forty-one eyes of 41 cases with different kinds of macular traction or macular pucker, including 12 cases of idiopathic epimacular membrane, 10 cases of macular hole complicated with epimacular membrane and 19 cases of macular pucker after reattachment surgery (no diabetic retinopathy or retinal detachment) were treated with vitreous surgery. The issue of this operation is to find out underlying fissures (an under-bridge) or weakly adherent sites between a membrane and the retina. Using flute-needle, intraocular pick, spade and vitreoretinal forceps, we can separate and peel the proliferative membrane and the posterior cortical vitreous from the retinal surface completely. RESULTS: In 41 cases, epimacular membrane was peeled successfully, and the normal anatomic structure of the macula was restored. Four cases had small amount of hemorrhage in the superficial layer of the retina in and after the operation, and all the other cases did not have any complications, such as retinal tear or detachment. Thirty-eight cases obtained higher visual acuity, and in 3 cases the visual acuity maintained at their pre-operative level. Metamorphopsia was significantly improved, even disappeared. The follow-up period was 3 months to 4 years. CONCLUSIONS: The therapeutic effects of vitreous surgery for treatment of various kinds of proliferative maculopathy are reliable. After the surgery, the visual acuity can be elevated, the symptoms can be ameliorated and the complications can be effectively controlled. PMID- 11864413 TI - [Evaluation of treatment for retinal detachment in juveniles]. AB - OBJECTIVE: To assess the effectiveness of different surgeries for retinal detachment in juveniles. METHODS: A retrospective analysis of the clinical therapeutic effects of various surgical treatments on 108 cases (116 eyes) in the age range of 3 - 14 years (mean, 9.6 years) was performed. RESULTS: Twenty-one eyes underwent scleral buckling procedure, and the retina was reattached in 19 eyes primarily. Fourteen eyes obtained post-operative visual acuity >or= 0.02. Ninety-seven eyes underwent vitrectomy, 58 (59.8%) eyes achieved retinal reattachment primarily and 83 eyes (85.6%) achieved retinal reattachment finally. Among them, retinal reattachment occurred in 7 of 11 gas-filled eyes, and the post-operative visual acuity >or= 0.02 in 5 eyes; and the reattachment occurred in 51 (57.3%) of 89 silicone oil-filled eyes, requiring only the primary repair. Of the silicone oil-filled eyes, 76 (85.4%) eyes achieved retinal reattachment finally. Fifty-nine (66.3%) silicone oil-treated eyes had visual acuity >or= 0.02 compared with 45.5% in long-acting-gas-treated eyes. CONCLUSIONS: Scleral buckling is a beneficial method in treatment of retinal detachment in juveniles. Vitrectomy should be considered for complicated retinal detachment. There is an advantage favoring silicone oil tamponade in achieving retinal reattachment. Retinotomy is a useful procedure for the retinal reattachment in eyes with severe proliferative vitreoretinopathy, however, it is necessary to be careful in selection of cases to undergo such a procedure. PMID- 11864414 TI - [Hypotony following vitreoretinal surgery]. AB - OBJECTIVE: To evaluate the incidence and risk factors of hypotony in patients who have undergone vitrectomy for complex retinal detachment. METHODS: The medical records of 142 patients with complex retinal detachments who underwent vitrectomy were reviewed. Hypotony was defined as a persistent intraocular pressure (IOP) of 5 mmHg or less for 6 months in the follow-up. RESULTS: Among this 142 patients, hypotony occurred in 16 (11.3%) in 1 year postoperatively. The rate of hypotony in the patients with traumatic retinal detachment was 18.9%, and 6.7% in the patients with rhegmatogenous retinal detachment, the difference being statistically significant (chi(2) = 4.89, P < 0.05). The rate of hypotony in aphakia was 20.9%, and 7.0% for phakia, the difference being statistically significant (chi(2) = 5.67, P < 0.05). The rate of hypotony in the patients with proliferative vitreoretinopathy (PVR)-C class was l.8%, and l5.1% in the patients with PVR-D class, the difference being statistically significant (chi(2) = 3.97, P < 0.05). The rate of post-operative hypotony in the patients with preoperative hypotony was 21.9%, and. 6.9% in the patients without preoperative hypotony, the difference being statistically significant (chi(2) = 5.16, P < 0.05). CONCLUSIONS: Intraocular hypotony is a complication of vitrectomy for complex retinal detachment. The risk factors for the post-operative hypotony include trauma, aphakia, preoperative hypotony and severe PVR. PMID- 11864415 TI - [Diagnostic and differential diagnostic potential of mitochondrial DNA assessment in patients with Leber's hereditary optic neuropathy]. AB - OBJECTIVE: To study the primary mutations of mitochondrial DNA (mtDNA) associated with Leber's hereditary optic neuropathy (LHON) in patients with optic neuropathy. METHODS: Seventy-nine patients with a variety of bilateral optic neuropathy were examined. Mutations at np 3,460, np 11,778 and np 14,484 of mtDNA were tested by PCR-restriction fragment length polymorphism technique to detect DNA in peripheral blood. The samples were taken from 16 cases of clinically diagnosed LHON, 44 cases of suspected LHON, two cases of alcohol amblyopia, four cases of multiple sclerosis, five cases of autosomal dominant hereditary optic atrophy, 4 cases with primary open-angle glaucoma, three cases of spinocerebellar degeneration, and one case of ethambutol-induced optic neuropathy. RESULTS: The mutation at np 11,778 was identified in 31 cases (39.2%), consisting of all the 16 clinically diagnosed LHON cases, thirteen cases (29.5%) of the suspected LHON, and the two cases of alcohol amblyopia. The remaining 48 cases were negative for mtDNA mutations at np 3,460, np 11,778, or np 14,484. CONCLUSION: Assessment of mtDNA provides a useful diagnostic aid in confirming and excluding the diagnosis of LHON, particularly useful in cases without a family hereditary history and cases with cause unknown bilateral optic neuritis. PMID- 11864416 TI - [Culture and identification of fetal human retinal photoreceptors]. AB - OBJECTIVE: To investigate the methods of isolation and culture of fetal human retinal photoreceptors. METHODS: Fetal human retinal photoreceptors were obtained by enzyme gradation digestion, cultured in media conditioned by retinal pigment epithelial cells (RPE-CM), identified by morphologic observation and immunohistochemically stained with rod specific opsin 4D2 (Rho 4D2) antibody. The digested retinal tissues were tested by hematoxylin-eosin (HE) staining. RESULTS: Enzyme gradation digestion could obtain relatively pure retinal photoreceptors. These cells were stained positively for Rho 4D2 antibody, and survived in RPE-CM for a long time. CONCLUSION: Fetal human retinal photoreceptors obtained by enzyme gradation digestion can survive in RPE-CM for a long time and express rod specific opsin. This culture system will provide a source for human retinal photoreceptor transplantation and further studies of photoreceptors in vitro. PMID- 11864418 TI - [Effect of high-vaccum-manual-chop technique in phacoemulsification]. AB - OBJECTIVE: To study the effect of high-vaccum-manual-chop technique to reduce complications related to phacoemulsification. METHODS: The correlation between the relative energy complex parameter (RECP) and the postoperative corneal edema rate (CER) in manual-chop and phaco-divide groups was statistically studied. RESULTS: The intraoperative mean relative ultrasound power and the mean time of ultrasound phacoemulsification were increased with the increase of the lens nucleus hardness. A significant difference existed between the mean time of the ultrasound phacoemulsification of the two groups (t = 3.1467, P < 0.01). The comparison between the mean relative ultrasound power of the two groups has no significant difference (t = 0.1874, P > 0.05). Positive correlation was observed between RECP and CER (r = 0.97, P < 0.01). CONCLUSION: High-vaccum-manual-chop technique may reduce the postoperative complications related to phacoemulsification, and RECP can be taken as a reliable index to evaluate the safety of phacoemulsification. PMID- 11864417 TI - [Preparation of genetically engineered retinal pigment epithelial cells capable of secreting nerve growth factor]. AB - OBJECTIVE: To develop genetically engineered retinal pigment epithelial (RPE) cells which are able to express bioactive nerve growth factor (NGF) steadily and efficiently and to combine gene therapy with retinal transplantation techniques to treat retinal degeneration. METHODS: Replication-deficient retrovirus-plasmid recombinant pLXSN-NGF was constructed via subclone and clone, and then packaged into pT67 cells to obtain virion containing a secretable form of NGF gene. We evaluated titer of the retroviral recombinant with NIH 3T3 cells and then transduced NGF gene into cultured RPE cells by the retroviral recombinant. We also analyzed the expression of NGF gene in transduced and untransduced RPE cells via reverse transcription (RT)-PCR and Western blot. The bioactivity of expressed NGF were evaluated by stimulating PC 12 cells with conditioned medium from transduced and untransduced RPE cells. RESULTS: The titer of replication deficient retroviral recombinant reached 1.2 x 10(7) cfu/ml. NGF protein and mRNA were detected in the conditioned medium from transduced RPE cells, but not from the normal RPE cells. Marked neurite process outgrowth from PC 12 cells was observed after being stimulated with the conditioned medium from transduced RPE cells. CONCLUSIONS: RPE cells are able to be transduced by replication-deficient retrovirus with high efficiency, and secret bioactive NGF at a high level, which might be an ideal candidate for engineered cells. PMID- 11864419 TI - [Comparison of nerve fiber layer thickness between optical coherence tomography and histomorphometry in glaucomatous monkey eyes]. AB - OBJECTIVE: To correlate the retinal nerve fiber layer (NFL) thickness measured by optical coherence tomography (OCT) in vivo with histomorphometric measurement. METHODS: Four laser-induced glaucomatous eyes of 4 cynomolgus monkeys were studied by OCT scanning a circular area 2.4 mm in diameter around the optic nerve head. The 4 eyes had early or advanced glaucoma. After final OCT measurement, the monkeys were sacrificed and the eyes enucleated. Routine serial section preparations were made, stained with hematoxylin and eosin, and the NFL thickness of 526 sections measured with the aid of a computerized image analysis system was compared with that at the points coincident with the OCT scanned. RESULTS: The mean OCT and histomorphometric NFL thickness in glaucoma were (72.2 +/- 54.5) microm and (61.1 +/- 44.2) microm, after the correction of dehydration 15% (70.3 +/- 51.1) microm respectively. Student's t-test showed that the difference was insignificant (t = 1.895, P > 0.05). CONCLUSIONS: The NFL thickness measured by OCT in vivo correlates well with that by histomorphometry, and the measurement is accurate. PMID- 11864420 TI - [The retinal nerve fiber layer defect and its related clinical features in early primary open-angle glaucoma]. AB - OBJECTIVE: To investigate the distribution of diffuse and local retinal-nerve fiber-layer defects (RNFLD) and their clinical features in early primary open angle glaucoma (POAG) and normal-tension glaucoma (NTG). METHOD: The types of RNFLD in 81 cases with POAG and 70 cases with NTG were observed, the distribution of diffuse and local RNFLD were analyzed, and the maximum untreated intraocular pressure (IOP) and the history of optic disc hemorrhage between the two types of RNFLD in the two glaucomatous groups were compared. RESULTS: RNFLD appeared in 78 of 81 cases with POAG, 50 of them were diffuse and 28 were local; 27 of diffuse and 43 of local RNFLD were found in 70 cases with NTG. The IOP was (28.78 +/- 5.41) mm Hg in the diffuse and (26.2 +/- 5.2) mm Hg in the local type in POAG (t = 2.04, P < 0.05), and (19.2 +/- 1.9) mm Hg and (17.8 +/- 2.0) mm Hg, respectively in NTG (t = 2.94, P < 0.05). The incidence of disc hemorrhage was 10/78 and 18/70 in POAG and NTG, respectively (chi(2) = 4, P < 0.05). That was 3/50 in the diffuse and 7/28 in the local type in POAG (chi(2) = 4.22, P < 0.05), and 3/27 and 15/43, in NTG respectively (chi(2) = 4.91, P < 0.05). There were no statistic differences in the diffuse type between the POAG and NTG groups (chi(2) = 0.15, P > 0.05), and neither were in the local type (chi(2) = 1.31, P > 0.05). CONCLUSION: The diffuse RNFLD tends to be distributed in POAG, while the local RNFLD tends to be distributed in NTG. The maximum IOP in the eye with diffuse RNFTD is higher than that in the eye with local RNFLD, either in POAG or NTG group. The occurrence of disc hemorrhage is higher in the eye with local RNFLD than that in the eye with diffuse RNFLD in the two groups respectively. There are differences in clinical features between the two types of RNFLD in early POAG and NTG. PMID- 11864421 TI - [The characteristics of bcl-2 and PCNA expression in the lens epithelium of human being]. AB - OBJECTIVE: To investigate the characteristics of bcl-2 (gene inhibiting apoptosis) and proliferating cell nuclear antigen (PCNA) expression and their relation to the proliferation of lens epithelium. METHODS: The expression of bcl 2 and PCNA were detected by immunohistochemical streptavidin-peroxidase (SP) method. RESULTS: The expression of bcl-2 and PCNA in the lens epithelium of fetus and children was higher than that of the elderly, especially in the region of the lens equator. CONCLUSION: The over-expression of bcl-2 and PCNA in the lens epithelium of fetus and children suggests that bcl-2 and PCNA might be related to the development of cataract. PMID- 11864422 TI - [Histological and ultrastructural studies of extraocular muscle proprioceptor in concomitant strabismus]. AB - OBJECTIVE: To investigate the morphological change of extraocular muscle proprioceptor (EMP) in concomitant strabismus and study its pathogenesis. METHODS: The samples were obtained from the extraocular muscles of patients with concomitant strabismus and enucleated eyeballs. After being stained, the samples were examined under light microscopy (LM) and transmission electron microscopy (TEM) to count the number of the mitochondria in each sample in an area of 4,000 mm(2) of the axon under the same amplification. RESULTS: Under light microscopy, at the myotendinous junction of the extraocular muscle, the EMP nerve fibers were ended at the initial site or the bilateral sides of a nearby single extrafusal fiber. Under electron microscopy, the general architecture of the receptors in concomitant strabismus was completely disorganized. The nerve component in them had disappeared. The number of the mitochondria in 4,000 mm(2) of the axon in the experimental group was significantly lower than that in the control group (P < 0.05). CONCLUSION: The results indicate that in concomitant strabismus, the reception and transmission of proprioceptive information are abnormal, and the morphological EMP disturbance plays an important role in the pathogenesis of concomitant strabismus. PMID- 11864423 TI - [A clinical observation of preserved donor tarsal plate transplantation for repair of tarsus defect]. AB - OBJECTIVE: To investigate the effect of preserved donor tarsal plate transplantation on the repair of the tarsus defect at the time of eye lid tumor removal. METHODS: Donor tarsal plates preserved in pure glycerin were used for the repair of tarsus defects of 58 cases after the complete removal of the eye lid tumors. The grafts were covered with rotated or slid myocutaneous and conjunctival flaps. The associations of the effect with the blood supply, with the size and the preserved time of the donor tarsal plates, as well as with the follow-up time were analyzed by chi(2) test. RESULTS: Of the 43 cases whose plates were fully covered with conjunctival flaps, new blood vessels began growing into the donor tarsal plates one week after the operation. The grafts were vascularized one month later. The donor tarsal plates of 15 cases were only partly covered with conjunctival flaps. As a result, the grafts were vascularized in 2 -- 3 months, and there was dissolution to varying degrees at the surface of the exposed parts of the plates. The effects of the 43 cases with good blood supply were significantly better than the effects of 15 cases with poor blood supply (chi(2) = 5.81, P < 0.05). The sizes of the donor tarsal plates ranged from 5 x 10 mm(2) to 8 x 29 mm(2). Eleven cases were equal or shorter than half of the original sizes of donor tarsal plates, 34 cases were subtotal and 13 cases were total plates. The differences of the sizes led to different results (chi(2) = 6.28, P < 0.05). The preserved times of donor tarsal plates were from 1 week to 2.5 years. However, the results were similar in all cases (chi(2) = 0.99, P > 0.05). All the patients were followed up for 6 months to 10 and half years (mean, 1.8 years). The effects had no statistical significance (chi(2) = 0.85, P > 0.05). The cilia of the donor tarsal plates were preserved in 3 cases, but all of them were totally lost during post-operative one month. After operation slight entropion occurred in 8 cases, slight ectropion in 3 cases, lagophthalmos for 2 - 3 mm in 7 cases, notch of the palpebral margin in 5 cases and thinness of the donor tarsal plates in 4 cases, but the functional and aesthetic results were normal. Thirty-one cases were cured, 11 cases acquired tangible results and 12 cases were improved. The total effective rate was 93.1%. CONCLUSIONS: The more blood supplies of the donor tarsal plates, the better the effects. So we must make all our efforts to improve the blood supplies of the donor tarsal plates. The effects were associated with the sizes of the plates, but not associated with the preserved times of the grafts and the follow-up times. Preserved donor tarsal plate transplantation for the repair of tarsus defect is a relatively simple operation, the rejection is slight and the result satisfactory, thus it is an ideal plastic material. PMID- 11864424 TI - [Experimental study and application of extracellular matrix of conjunctiva]. AB - OBJECTIVE: To investigate the characteristics and application of extra-cellular matrix (ECM) of conjunctiva. METHODS: Pieces of conjunctival ECM were made, and then the conjunctival defect was repaired with that in an experimental group. Control group 1 received the heterogeneous conjunctiva, and in control group 2, the refrigerated ECM was used for transplantation. All of them were photographed and examined by light microscope, electron microscope (EM), immunohistochemical examination and lymphocyte toxicity test. RESULTS: There were blood vessels growing into the grafts on the 3rd day in the experimental group. The grafts became mildly congestive and similar to normal conjunctival ones in the 4(th) week. But in control group 1, the grafts became white and necrotic in the second week; the conjunctiva was congestive, cicatricial and hyperplastic in the 4(th) week. Under the light microscope, the epithelium covered the most part of the region with the graft in the 2nd week, covered completely in the 4(th) week, and its appearance was as an approximately normal conjunctival epithelium in the experimental group. But the epithelium became ulcerative and a lot of lymphocytes infiltrated in the 2nd week in control group 1. Under the EM, the line of demarcation between the covering region of the regenerative conjunctival epithelium and the naked ECM was quite clear in the 1st week in the experimental group. And the ultrastructure of regenerative conjunctival epithelium was basically the same as an normal one in the 8(th) week. Both before and after transplantation, the immunohistochemical examinations verified that ECM was type I and IV collagen. The lymphocyte toxicity test showed that no obvious humoral immunoreaction existed. CONCLUSION: Conjunctival ECM has the characteristics of tissue activity and low antigenicity. It is an ideal conjunctival succedaneum in the plastic surgery of conjunctiva. PMID- 11864425 TI - [Clinical and histopathologic analyses of after cataract in rabbit eyes having undergone three different cataract surgeries]. AB - OBJECTIVE: To discuss the surgical method which can efficiently prevent the occurrence of after cataract. METHODS: Eighteen experimental rabbits were randomly divided into 3 groups. Group A was treated with phacoemulsification, group B with phacoemulsification combined with posterior continuous curvilinear capsulorhexis (PCCC) and Group C phacoemulsification combined with PCCC and anterior vitrectomy. Post-operatively, all the surgical eyes were checked with slit lamp microscope and ophthalmoscope. At the post-operative 3 months, histopathologic examinations were performed. RESULTS: There were no post operative complications, such as retinal detachment, cystoid macular edema, etc. There were Elschnig's pearls and fibers in the peripheral area of the posterior capsule, and the amount was about the same in the 3 groups. There were Elschnig's pearls and fibers at the central area in group A. The fibrous membrane grew from the edge of the PCCC in group B. But in group C, the central area of the posterior capsule was clear, and there was no obvious pathological changes. CONCLUSION: Phacoemulsification combined with PCCC and anterior vitrectomy is a safe cataract surgical method which can effectively prevent the formation of after cataract. PMID- 11864426 TI - [A pathological study of lens subcapsular epithelial proliferation]. AB - OBJECTIVE: To observe the histopathological changes of lens subcapsular epithelial proliferation and analyze their etiology. METHODS: Sixty-four enucleated eyes with lens subcapsular epithelial proliferation were observed with light microscopy and analyzed retrospectively. Among all the cases of lens subcapsular epithelial proliferation, there were 36 ones enucleated because of trauma, 10 absolute glaucoma, 6 corneoscleral staphyloma, 5 intraocular tumor, 2 after cataract extraction, 4 endophthalmitis, 1 cataract combined with ocular atrophy (diabetes type II). RESULTS: Proliferated lens subcapsular epithelial cells were atypical, irregularly arranged, and extended to the posterior subcapsule. In cases with lens capsular rupture or partial absence, there were subcapsular fibro-connective tissue metaplasia, scar tissue formation, cortical collapse, local liquefaction and calcification. CONCLUSION: Lens subcapsular epithelial proliferation is caused by some intraocular pathological conditions. In cases with endophthalmitis and ocular trauma, its severity is the most serious. PMID- 11864427 TI - [A survey and treatment of blindness in Gejiu City of Yunnan Province]. AB - OBJECTIVE: To calculate the percentage of blindness in the mass and evaluate the current conditions of treatment for cataract. METHODS: In accordance with the national unified standards and procedures, a clue survey of the people aged under 40 years and a general survey of the people over 40 years in Gejiu city, Yunnan province were carried out. In the mean time, operations were performed for the patients with cataract. RESULTS: Of the total 361 214 persons being surveyed, there were binocular blindness in 1 037 cases (0.29%) and monocular one in 983 cases (0.27%). Of the cases with binocular blindness, 593 cases (57.18%), and of the monocular blindness, 617 cases (62.77%) could be treated. The leading causes of 1037 cases with binocular blindness were cataract (51.98%), injuries of the eye (19.96%), corneal opacity (16.00%), glaucoma (10.03%), refractive error and other ocular diseases (2.03%). The average prevalence rate in the female cases was higher than that of male cases (chi(2) = 53.48, P < 0.01). Blindness was associated with older age There were variations in the prevalence rates in different aboriginals and in cases with different occupations. Cataract was the main disease to cause both the monocular and binocular blindness. Among the patients with cataract 539 cases (90.98%) of binocular blindness and 500 cases (81.03%) of monocular blindness were curable. The cataract operation was performed on 738 cases with cataract, including 396 cases with binocular blindness and 342 cases with monocular blindness. At postoperative 3 months, the rate of the best corrected visual acuity (eye sight > 0.05) was accounted for 97.73%, the rate of the eye sight > 0.3, 86.36% and the rate of wearing glasses (including implantation of intraocular lens), 85.60%. CONCLUSION: The prevalence of blindness is associated with the aboriginal, occupation, age and the sex. The leading cause of blindness is cataract. So the surgical restoration of vision in cases with cataract is the main measure in the prevention of blindness. PMID- 11864428 TI - [Primary research about reconstruction of cornea in three-dimensional collagen gel in vitro]. AB - OBJECTIVE: To research the reconstruction of cornea in three-dimensional culture. METHODS: Using primary cultured corneal epithelial cells, stromal cells and endothelial cells rebuilt a corneal structure in three-dimensional collagen gel reconstruction system. The artificial corneal transparency was examined by a slit lamp microscope. The biological characters of cells were tested by routine pathologic and transmission electron microscopic technology. RESULTS: The reconstructed corneal tissue formed three layers. The epithelial cell layer was two or three cell thickness. The endothelial cells formed a single layer. The stromal cells were diffuse in distribution in the collagen gel. The cells had proliferative ability and capability of producing collagen. CONCLUSION: The regular and transparent corneal structure can be rebuilt by tissue engineering technology. PMID- 11864429 TI - [Preparation of green fluorescent protein retrovirus and its application in mediating gene transfer into retinal pigment epithelial cells]. AB - OBJECTIVE: To determine whether human retinal pigment epithelial (RPE) cells can be infected by retrovirus and modified by retrovirus-mediated gene transfer. METHODS: Recombination retroviral vector pLNCX-GFP (green fluorescent protein, GFP) was generated by inserting 780 bp GFP cDNA fragment into the MCS site of pLNCX. pLNCX-GFP was transfected into ecotropic packaging cell line PhiX-Eco and amphotropic packaging cell line PhiX-Ampho and PA317. Retroviral titer was tested by counting GFP expression of NIH3T3 cells. Then RPE cells were infected by using GFP retrovirus-containing supernatant. RESULTS: GFP was expressed and retrovirus was produced upon pLNCX-GFP being transfected into packaging cell line. The GFP retrovirus was able to infect primary cultured human RPE cells and immortalized RPE cell line. CONCLUSION: The retrovirus can introduce a foreign gene into RPE cells efficiently, thereby it can be used as an important tool to deliver gene into RPE for therapy of fundus diseases. PMID- 11864430 TI - [Reconstruction of corneal stroma using tissue engineering technique]. AB - OBJECTIVE: To reconstruct new corneal stroma in order to provide a basis for future studies on the reconstruction of cornea. METHODS: The cultured corneal stromal cell-PGA complex was grafted to the subdermis of a female balb/c nude mouse. Six weeks after grafting, the new reformed tissue was fixed for histologic examination and transmission electron microscopy (TEM). At the same time, the new deposited collagen fibril diameter was measured. RESULTS: The reconstructed corneal stromal layer was formed in a waved, crossed-web structure, which was similar to the normal corneal stroma. Cell deposited collagen fibrils were similar in diameter to those seen in normal corneal stroma. CONCLUSIONS: The corneal stroma can be reconstructed by using tissue engineering technique. The reconstructed tissue which is similar to the normal cornea in key characteristics, including morphology and histologic structure provides an ideal experimental model for corneal physiological, pathologic, toxic and drug efficient testing and for future development of the tissue for implantation. PMID- 11864431 TI - [A gene study on multidrug resistance of retinoblastoma]. AB - OBJECTIVE: To investigate the expression of multidrug resistance gene (MDR1) and multidrug associated protein (MRP) gene in the retinoblastoma (Rb) cell line HXO RB(44), and study the mechanism of multidrug resistance (MDR) phenotype occurring in retinoblastoma. METHODS: Reverse transcriptase polymerase chain reaction (RT PCR) was employed to detect the transcription of MDR1 and MRP gene in the HXO RB(44) cell line, and immunohistochemical technique was used to detect their protein products P-glycoprotein and multidrug resistance associated protein (P190). RESULT: Both MDR1 and MRP genes were expressed in HXO-RB(44) cell line, as well as P-glycoprotein and P190 were over-expressed (96% - 97%) in this cell line. CONCLUSION: Over-expression of MDR1 and MRP genes is significantly involved in the mechanism of MDR phenotype in retinoblastoma. PMID- 11864432 TI - [A study on effect of basic fibroblast growth factor on human limbal stem cell proliferation cultured in low calcium medium]. AB - OBJECTIVE: To study how to obtain relatively pure stem cells and the effects of bFGF on the human limbal stem cell proliferation in order to set up the foundation of cultured limbal stem cells for treatment of ocular surface and tear diseases. METHODS: Limbal button inoculation was used in low calcium culture. Monoclonal antibody AE5 was used to identify the stem cells. To examine the effects of bFGF (1 - 100 ng/ml) on stem cell proliferation, the technique of digital camera and computer image analysis system were used. RESULTS: Cultured cells were mostly corneal stem cells. This was the first time to combine the method of obtaining basal cell layer and culture with low calcium medium, which could harvest relatively pure and undifferentiated stem cells in localization and nature. The basic FGF (1 - 100 ng/ml) promoted cultured stem cells to proliferate significantly (P < 0.001). CONCLUSIONS: Limbal stem culture provides resources of stem cells. It is very useful in treatment of ocular surface and tear diseases. The basic FGF plays an important role in proliferation of stem cells in graft. Computer image analysis system is an effective and simple way for evaluating the proliferation of a primary culture. PMID- 11864433 TI - [Study on the association of pathologic myopia with HLA-DQB1 gene]. AB - OBJECTIVE: To investigate the association of pathologic myopia (PM) with HLA-DQB1 alleles in order to study the pathogenesis of PM. METHODS: The genome DNA of 66 patients with PM was extracted from the blood. The second exon of the HLA-DQB1 gene was amplified by polymerase chain reaction (PCR), and individual PCR products were digested by allele specific restriction enzymes of Hae III, BssH II, Apa I, BsaH I, Hae II, Hpa II, Rsa I, Bsp 1286 I for typing. Genotype was determined by restriction fragment length polymorphism (RFLP) pattern. The frequency of 16 alleles of HLA-DQB1 was detected and compared with that of the healthy control. RESULTS: The frequencies of HLA-DQB1 * 0301 and * 0303 were significantly higher in the patients with PM (P < 0.0001), while that of HLA-DQB1 * 0601 and * 0602 were significantly lower (P < 0.0001) than that of the control group. CONCLUSION: HLA-DQB1 * 0301, * 0303 alleles are susceptible alleles, and possibly they are the pathogenic genes, * 0601, * 0602 are resistant alleles, and may be they possess the protective property. PMID- 11864434 TI - [A study of adhesion molecule expression in extraocular muscles of Graves' ophthalmopathy]. AB - OBJECTIVE: To investigate the expression of intercellular adhesion molecule-1 (ICAM-1) and vascular adhesion molecule-1 (VCAM-1) in extraocular muscles (EOM) of the eyes with Graves' ophthalmopathy (GO). METHODS: Seventeen EOM specimens obtained from 16 severe patients with GO and their cryostat sections were observed by double labeling immunohistochemical methods. RESULTS: The expression of ICAM-1 was observed in interstitial and perimysial connective tissue surrounding EOM fibers, mononuclear cells, fibroblasts and vascular endothelial cells. Only were vascular endothelial cells stained for VCAM-1, and ICAM-1 and VCAM-1 immunoreactivities were not detected in EOM cells. CONCLUSIONS: The findings indicate that ICAM-1 and VCAM-1 might be involved in the development of GO, and fibroblasts, vascular endothelial cells and mononuclear cells might play important roles in the autoimmune reaction. PMID- 11864435 TI - [An experimental study on subconjunctival interleukin-1 receptor antagonist for promotion of corneal transplant survival]. AB - OBJECTIVE: To determine whether the subconjunctival application of interleukin-1 receptor antagonist (IL-1ra) can prolong the corneal graft survival in the rat model of orthotopic penetrating keratoplasty. METHODS: For all experiments, F344 corneas were transplanted into LOU (major histocompatibility-disparate) eyes. Experimental groups received subconjunctival injection of 50, 100 and 200 microg IL-1ra respectively, and the control group received the same volume of 0.9% normal saline instead for consecutive 2 weeks. All transplants were evaluated for 4 weeks after surgery for signs of rejection. RESULTS: The mean survival time (MST) of the grafts of the experimental groups was increased significantly (t = 0.00, P < 0.01) in comparison with the control group. The MST of the IL-1ra 200 microg group was increased significantly than that of the IL-1ra 50 microg group (t = 0.00, P < 0.01). Furthermore, the IL-1ra-treated grafts had significantly less corneal inflammation, infiltration, lower levels of opacity, edema, neovascularization and rejection index compared with the control group. CONCLUSIONS: Subconjunctival treatment of IL-1ra has a significantly positive effect on promoting corneal allograft survival. And its effect is dosage dependent. PMID- 11864436 TI - [The clinical effect of non-penetrating trabecular surgery with reticulated sodium hyaluronate implant for treatment of primary open-angle glaucoma]. AB - OBJECTIVE: To evaluate the clinical effect of non-penetrating trabecular surgery (NPTS) with reticulated sodium hyaluronate implant in primary open-angle glaucoma (POAG). METHODS: NPTS with reticulated sodium hyaluronate implant was performed on 27 eyes of 25 patients with POAG. The procedure consisted of excising a deep scleral tissue including the external wall of the Schlemm's canal under a scleral flap without opening the anterior chamber, as the inner wall of the canal was left in place. Then placing a 3.0 mm x 4.5 mm x 0.5 mm or 3.5 mm x 3.5 mm x 3.5 mm reticulated sodium hyaluronate implant was placed under the flap, so that the aqueous humor may filter through the thin layer of trabeculocorneal membrane spontaneously without forming a flat chamber. The postoperative intraocular pressure (IOP), inflammation and filtering bleb were analyzed. All of patients undertook ultrasound biomicroscopy (UBM) and gonioscopy to evaluate the surgical site on the postoperative 2 weeks, 3, 6 and 9 months. The mean follow-up was (6.61 plus minus 2.47) months. RESULTS: The IOP decreased from a mean preoperative value of (26.36 +/- 9.02) mm Hg (1 mm Hg = 0.133 kPa) to a mean postoperative value of (14.18 +/- 3.51) mm Hg (t = 6.875, P < 0.05). The number of anti-glaucomatous medications, topical or systemical, was reduced from (2.96 +/- 1.43) sorts preoperatively to (0.77 +/- 1.07) sorts postoperatively. The visual acuity remained stable (no statistical difference with chi(2) test). Six eyes had elevated IOP after operation and were controlled by eyedrops or trabecular puncture with ND: YAG laser. A slight hyphema occurred in 4 eyes with small puncture during operation. The complications such as flat chamber, inflammation and choroidal detachment were not observed. The UBM showed that the sodium hyaluronate implant degraded and a transparent liquid space existed under the scleral flap in all of patients at postoperative 3 months. The gonioscopy showed that at the surgical site the thinner and semitransparent trabecular membrane and changes of a transparent cavity could be seen in 23 eyes. In the other eyes, at the site the trabecula was translucent. The filtering blebs were formed in 8 eyes. CONCLUSION: Non-penetrating trabecular surgery with reticulated sodium hyaluronate implant can effectively lower the IOP and reduce the sorts of anti-glaucomatous medications. The visual acuity may remain unchanged, and no serious complications commonly seen in the traditional trabeculectomy occur. The procedure is a new simple and effective one for the treatment of POAG. PMID- 11864437 TI - [The scleral buckling combined with argon laser photocoagulation for retinal detachment]. AB - OBJECTIVE: To investigate the effects and indications of postoperative photocoagulation with argon laser instead of intraoperative cryotherapy for closing the retinal breaks during the scleral buckling operation in rhegmatogenous retinal detachment (RD) METHODS: An analysis of clinical data in 35 RD cases (38 eyes) was conducted retrospectively. These cases were treated by argon laser photocoagulation after the scleral buckling surgery from December, 1999 to April, 2000 in our hospital. RESULTS: Thirty-eight eyes were followed-up from 3 to 6 months. In 36 of 38 eyes, the retina was reattached after surgery (94.7%). Postoperatively, thirty-one eyes (86. 1%) had a visual acuity (VA) of >or= 0.05 and the best VA was 1.0. CONCLUSIONS: The method using postoperative photocoagulation instead of cryotherapy during scleral buckling surgery can simplify the surgical manipulations, alleviate the vessel damage of the retina and uvea and reduce the postoperative complications. It is a practicable way to treat RD. Surely, photocoagulation can not totally replace cryotherapy. Proper selection of indications and avoidance of complications of photocoagulation can elevate the successful rate of reattachment of RD, and promote the recovery of visual function. PMID- 11864438 TI - [A clinical investigation on the relationship between nitric oxide and elevation of intraocular pressure in glaucoma]. AB - OBJECTIVE: To investigate the role of nitric oxide (NO) in the process of intraocular pressure (IOP) elevation in glaucoma. METHODS: The levels of NO in plasma and in aqueous humor which were obtained from 16 patients with acute angle closure glaucoma (10 female, 6 male), 29 cases with cataract (13 female, 16 male) and 30 normal volunteers (13 female, 17 male) were detected by high performance liquid chromatography and expressed with the total NO(2) and NO(3). RESULTS: It was found that the mean level of NO in the plasma in glaucoma group was (l,390.78 +/- 302.95) ng/L that was significantly lower than that in the normal group (2 969.37 +/- 862.85) ng/L (t = 2.853, P < 0.01). But in cataract group it was (4,234.89 +/- 1,495.23) ng/L that was significantly higher than that in the normal group (t = 4.471, P < 0.01). There was no significant difference in the level of NO in the aqueous humor between glaucoma group (2,052.03 +/- 694.74) ng/L and normal group (1,405.83 +/- 602.70) ng/L (t = l.862, P > 0.05). But in cataract group the mean level of NO (2, 454.67 +/- 655.13) ng/L in the aqueous humor was increased remarkably in comparison with that in the normal group (t = 2.752, P < 0.01). The rate of the mean content of NO in normal aqueous humor and plasma (aqueous/plasma) was 47.3%, and that in cataract group, 57.96% (P > 0.05). But in glaucoma group, the rate was 144.0%. CONCLUSION: The content of nitric oxide in the plasma and aqueous humor is related to the elevation of IOP in cases with acute angle closure glaucoma. PMID- 11864439 TI - [Phacoemulsification of the subluxated lens with cataract]. AB - OBJECTIVE: To study the efficiency of phacoemulsification in the treatment of subluxated lens with cataract. METHOD: Phacoemulsification combined with intraocular lens implantation was performed on 19 eyes of 18 patients. RESULT: The visual acuity in 18 eyes was improved to different degrees. The visual acuity in one eye was not improved due to the preoperative macular contusion. CONCLUSION: Phacoemulsification for treatment of subluxated lens with cataract may decrease many complications occurring in the past procedure. PMID- 11864440 TI - [Surgical treatment of congenital nuclear cataract]. AB - OBJECTIVE: To study the characteristics of congenital nuclear cataract and evaluate the therapeutic effects of extracapsular cataract extraction (ECCE) with or without intraocular lens (IOL) implantation in different age groups. METHOD: ECCE with or without IOL implantation was performed on 76 eyes of 38 cases with congenital nuclear cataract in different age groups. Under operative microscope, the characteristics of the opacity of the posterior capsule were described. The visual acuity and possible complications were followed up for a mean period of 6 months. RESULTS: The opacity of the posterior capsule was progressively developing, its diameter was similar to that of the opaque nucleus. The difference of the post-operative corrected visual acuity in different age groups was significant (chi(2) = 27.84, P < 0.005). The rates of the secondary cataract were negatively correlated to the age groups (r = -0.969 2, P < 0.05). In 2 eyes, slight iris capture occurred in 3 - 12 year group. CONCLUSIONS: Congenital nuclear cataract can combine with posterior capsular cataract. The etiology remains to be studied. The therapeutic effects of the congenital nuclear cataract mainly depend on the time of operation. The early treatment of post-operative complications in children is of primary importance. PMID- 11864441 TI - [Postoperative use of YAG laser for cataract surgery]. AB - OBJECTIVE: To study and analyze the postoperative use of YAG laser for cataract surgery, the method, effects and complications of laser posterior capsulotomy. METHODS: Four hundred and fifty-one eyes out of 4 600 eyes with phacoemulsification in a single horizontal groove manner were dealt with YAG laser for postoperative complications, and of them, 380 eyes had a posterior capsulotomy with a two-steps method: posterior capsular circular incision and push down of the capsular chip. Forty-two eyes were treated for anterior membrane on the intraocular lens (IOL) by circular resection and peeling it off. The other 29 eyes were treated for different complications. RESULTS: The rate of YAG laser treatment for postoperative complications was 9.8% and of them, the rate of posterior capsulotomy was 8.3%. The rate of eyes with posterior capsular opacification and one or more other complications was 14.2%. The mean energy of a single pulse for posterior capsular resection was (1.21 +/- 0.31) mJ, and the mean total energy was (52.44 +/- 14.62) mJ; the mean energy of a single pulse for push down of the capsular chip was (2.12 +/- 0.43) mJ, and the mean total energy was (50.88 +/- 10.32) mJ. The average naked visual acuity after posterior capsulotomy was increased to 0.58 from 0.32 before the laser treatment, and corrected visual acuity was 0.85. The energy of a single pulse was 1.0 to 1.4 mJ for anterior membrane resection of IOL, and the mean total energy was (56.28 +/- 32.74) mJ. No obvious damage or decentration of the IOL was found. After the treatment, the IOP in 95.3% of eyes was kept in normal range. Only did 2 eyes suffer from retinal detachment in follow-up. CONCLUSIONS: YAG laser therapy for postoperative complications is easy to control, safe, effective, and has less complications. Besides the posterior capsulotomy, some other complications also can be cured with it. PMID- 11864442 TI - [Clinical and MRI diagnosis of nasopharyngeal carcinoma with orbital spread]. AB - OBJECTIVE: To study the routes, clinical manifestations and MRI characteristics of nasopharyngeal carcinoma (NPC) with orbital spread. METHODS: Twenty (23 eyes) such cases were selected, including 6 (7 eyes) preradiotherapy cases and 14 (16 eyes) postradiotherapy cases. The magnetic resonance imaging (MRI) axial, coronal, sagittal routine procedures and gadolinium-diethylene triamine pentaacetic acid (GD-DTPA) contrast-enhanced scan were performed. Fatty-restraint scan was performed after enhancement only on 3 cases. The diagnosis in all the patients was proved by the biopsy from the nasopharynx. RESULTS: (1) Single orbit involvement was seen in 17 patients, 12 in the left and 5 in the right orbit. Bilateral orbit involvement was only found in 3 patients. (2) Major clinical manifestations of orbital involvement by NPC included: 13 cases (15 eyes) suffered from decrease of visual acuity, 9 cases (10 eyes) with exophthalmos, 11 patients (12 eyes) with ocular movement disorder, 7 cases (9 eyes) with diplopia and 5 cases (7 eyes) with blepharoptosis. (3) MRI appearances: There were 13 cases (15 eyes) with optic nerve involvement, 9 patients (11 eyes) with abnormalities in extraocular muscle, 8 cases (9 eyes) with retrobulbar tumor and 15 cases (17eyes) with orbital apex and orbital lamina bony involvement. (4) The three main routes for orbital involvement in NPC patients were as follows: (a) through cavernous sinus (the skull base) to superior orbital fissure. (b) through paranasal sinus (particularly the ethmoid sinus). (c) through pterygopalatine fossa to inferior orbital fissure. CONCLUSION: MR imaging is very useful to demonstrate the presence, location and extent of orbital involvement in NPC patients. PMID- 11864443 TI - [Experimental research of laser-induced collaterals in retinal vein occlusion]. AB - OBJECTIVE: To evaluate the feasibility and safety of establishing chorioretinal venous anastomosis in rabbit eyes with a model of branch retinal vein occlusion METHODS: By using laser photocoagulation, a chorioretinal venous anastomosis was created in rabbit eyes in which a branch retinal vein occlusion had previously been established photochemically. A similar attempt to create an anastomosis was made in the control eyes in which no branch vein occlusion was present. RESULTS: In the 21 eyes with the model of branch retinal vein occlusion, a chorioretinal venous anastomosis formed in 4 eyes within 3 to 5 weeks. In the control eyes, an anastomosis in one eye presented by 5 weeks, during follow-up of 2 to 5 months. The hemorrhage in choroid, retina and vitreous associated with laser photocoagulation were absorbed within 3 weeks, and no other severe complications were found. CONCLUSIONS: Chorioretinal venous anastomosis can be created by using laser photocoagulation. This technique is relatively safe. However, to improve the rate of successfully creating an anastomosis, more research work needs to be performed. PMID- 11864444 TI - [A study on susceptibility of different layers of rabbit retina to ocular hypertension]. AB - OBJECTIVE: To investigate the histological damage and its degrees of different layers of rabbit retina induced by ocular hypertension. METHODS: The intraocular pressure (IOP) was elevated by intracameral injection of 0.10 - 0.15 ml of 2% methylcellulose in sixteen gray rabbits. The rabbit eyes were enucleated, fixed, and paraffin-embedded. For histological evaluation, the slides were stained with hematoxylin-eosin, examined by light microscopic and computer automatic image analysis. RESULTS: The quantitative analysis indicated that all of the retinal layers have different degrees of histopathological damage. Among them, the retinal ganglion cells (RGCs) and retinal nerve fiber layer (RNFL) were most severely damaged, with the decrease rate of 74.3% and 61.9%, respectively. The decrease rates of other retinal layers were as follows: the retinal inner layer, 42.4%; the whole sensory layer of the retina, 35.0% and the outer layer, 24.2%. We also found a significant correlation between RGCs loss and RNFL thinning (r = 0.68, P < 0.01), demonstrating a strong correlation between IOP level and RNFL thickness (F = 8.97, P < 0.01). CONCLUSION: All of the anatomic layers of retina can suffer from degeneration and atrophy at different degrees. PMID- 11864445 TI - [Smoking, related cancers, and other diseases in shanghai: a 10-year prospective study]. AB - OBJECTIVE: To explore the relationship between smoking and related diseases among residents of Shanghai. METHODS: A cross-sectional study on current smoking status among 213,800 residents aged 20 yr and over in Shanghai urban, suburb and rural areas was carried out during the early 1980s. The residents in the urban area were followed up for 12 years (1983 - 1994) and those in suburb and rural areas for 11 years (1984 - 1994). A Poisson regression model was used to estimate relative risks RRs of certain diseases for smoking adjusted for age with their 95% confidence intervals by sex and area among persons aged 40 yr and over at the beginning of the study. RESULTS: In the urban area, the RRs of death for smoking were 1.48 and 1.62 in males and females, respectively and for cancer death were 2.20 and 2.00. Statistically significant elevated RRs were seen for cancers of the lung and liver in both males and females and for cancers of the oesophagus, stomach, pancreas and bladder only in males. Significantly higher RRs were also observed for cerebrovascular disease, chronic bronchitis and emphysema, and pulmonary heart disease. RRs lower than in the urban area were found in the suburb and rural areas. The population attributable risks (%, PAR) for death due to smoking for males were estimated by area. PARs for death were 20.9, 18.9 and 16.3 in urban, suburb and rural areas, respectively. For all cancers, PARs were 40.0, 34.5 and 34.2 and for lung cancer were 71.7, 59.2 and 64.7. CONCLUSION: Smoking was associated with the mortality of some common cancers, chronic bronchitis, and emphysema, pulmonary heart, and cerebrovascular diseases. PMID- 11864446 TI - [A case-control study on relationship between body mass index and lung cancer in non-smoking women]. AB - OBJECTIVE: To investigate the relationship between body mass index (BMI) and lung cancer in non-smoking women. METHODS: A population-based case-control study on lung cancer in non-smoking women was conducted during February 1992 to December 1993 in Shanghai. Totally, 504 cases of lung cancer of non-smoking women and 601 population controls were studied. RESULTS: The risk of lung cancer increased with the decrease of BMI. The odds ratio of lung cancer in non-smoking women was 1.95, as compared to controls with the highest quartile of their BMI, with a dose response pattern (P < 0.0002 with chi(2) test for trend), adjusted for age, schooling and income. Non-conditional logistic regression analysis was used to adjust for some main risk factors found in other studies carried out at home, such as passive smoking, tea drinking, eye irritation by cooking oil fume, kinds of cooking oils, pulmonary tuberculosis, family history of lung cancer, age of menarche, menstrual cycle and dietary nutrition (including intake of vitamins C and E, beta-carotene, total fat and calorie), was used to adjust. There still existed relationship between BMI and lung cancer in non-smoking women after adjustment for those factors. Stratification analysis showed that there was relationship mainly between BMI and lung adenocarcinoma. CONCLUSION: BMI may be a risk factor for lung cancer in non-smoking women. PMID- 11864447 TI - [Development of plasmid-based transgenic mice carrying with target gene xylE]. AB - OBJECTIVE: To develop a transgenic mouse model carrying with plasmid pUC118NX integrated into its genomic DNA for detecting mutagenesis. METHODS: DNA of plasmid UC118NX in target gene xy1E was injected microscopically into male protonucleus of 376 mouse spermatova, and 225 survival spermatova were transferred into the oviducts in both sides of 11 pseudopregnant female mice to develop their offspring. The genomic DNA in survival young mice were analyzed with polymerase chain reaction (PCR)-Southern blot, plasmid transformation test and endonuclease-digestion. RESULTS: Seven pseudopregnant mice got pregnant, and 29 offspring were delivered and 25 survived of which 18 were identified with positive for PCR-Southern blot (72%). The two stout male mice with intact integration of plasmid pUC118NX in their genome were finally chosen as founders to detect gene mutation in vivo and establish transgenic mouse lineages. CONCLUSION: Transgenic C57BL/6J mice integrated with plasmid pUC118NX into their genomic DNA have been successfully developed. PMID- 11864448 TI - [Effects of vitamins E, C and beta-carotene on DNA damage]. AB - OBJECTIVE: To investigate their effects on lymphocyte damage with supplementation of high-dose vitamins E, C and beta-carotene. METHODS: Healthy men aged 50 - 59 years were selected and randomized into the trial and control groups with 50 in each one. In the trial group, 25 mg of beta-carotene, 100 mg vitamin C and 280 mg vitamin E were given to each subject every day for 20 weeks. Whole blood was collected in both the trial and control groups and its lymphocytes separated. Damage to DNA was analyzed with a"comet" electrophoresis technique, and serum levels of vitamin C, beta-carotene and alpha-tocopherol were determined by high performance liquid chromatography. RESULTS: There was no significant difference in the proportions of spontaneous damage to DNA in lymphocytes between the trial and control groups (6.1% vs. 6.8%). But, proportions of damage to DNA in peripheral lymphocytes increased to 36.14%, 59.45, and 69.62%, respectively, after treatment with 30 micromol/L, 100 micromol/L and 300 micromol/L of H(2)O(2). CONCLUSION: Supplementation of vitamin E, C and beta-carotene could effectively reduce the damage to DNA caused by H(2)O(2). PMID- 11864449 TI - [The correlation of free radicals induced by cadmium toxicess with erythrocyte injury and band3 expression]. AB - OBJECTIVE: To study the correlation of hydrogen peroxide induced by cadmium ion with erythrocyte injury and band3 expression. METHODS: It was application immunoscanning electronmicroscopy and cytochemistry and EDX methods. RESULT: Cadmium ion not only induced a lot of hydrogen peroxide but also caused serious distortion of erythrocyte. CONCLUSION: Cadmium ion-induced free radicals lead to oxygen free radical damage of membrane and crytoskelecton of erythrocyte, affecting the stability and deformability and band3 expression in erythrocyte. PMID- 11864450 TI - [Fisher discriminant analysis for carcinogenic potency of aromatic amines]. AB - OBJECTIVE: To study the relationship between carcinogenicity and their structure and physicochemical parameters of aromatic amines. METHODS: Fisher discriminant analysis for carcinogenicity of aromatic amines in two different batches was conducted with estimation of parameters of physicochemical using central atom and functional groups in combination with structural parameters of polycyclic aromatic hydrocarbons (PAHs). RESULTS: The discriminant efficacy approximated to that of the two in a series of equations (I and III) established by Yuta, but inferior to the optimal equation II. CONCLUSION: The selected physicochemical parameters can be applied to QSAR study on carcinogenicity of most aromatic amines, with better results in Fisher discriminant analysis. PMID- 11864451 TI - [Effects of humic acid on lipid peroxidation in arsenosis prevalent areas]. AB - OBJECTIVE: To explore the causes of blackfoot disease in Taiwan. METHODS: Experiments to induce lipid peroxidation by arsenic and humic acid were made in vitro. Arsenic was determined for the water sampled from the arsenosis prevalent areas in Inner Mongolia and blackfoot disease prevalent areas in Taiwan and humic acid determined for the extracts of the coal rich in arsenic sampled from Guizhou Province in China by MDA-TBA colorimetry. RESULTS: Experiments found that humic acid could induce lipid peroxidation caused by sodium salt of unsaturated fatty acids in vitro in a weak and unstable manner, which could be promoted by 0.05 mmol/L of Fe(+ +) and not by 1 mmol/L As(2)O(3). The extent of lipid peroxidation caused depended on the sources, constituents and structure of humic acid samples, those from Inner Mongolia ranked the highest, those from Guizhou and Taiwan the medium and the commercial humic acid (Aldrich Co. in the US) the lowest. The experiments also found that lipid peroxidation caused by commercial humic acid presented a dynamic process in the existence of trace amount of iron, and humic acid could decompose the products of lipid peroxidation. CONCLUSION: The experiments suggested the relations of humic acid and arsenic to iron and other transition elements in the blackfoot disease prevalent areas and areas with an environment rich in arsenic and humic acid. PMID- 11864452 TI - [Studies on relationship between occupation and pregnancy outcome]. AB - OBJECTIVE: To study the effects of maternal occupation on reproductive outcomes, including pregnancy outcome and neonatal status, and complications of pregnancy and delivery. METHODS: Data were extracted from 12 224 obstetric records of eight hospitals in Beijing, and all the subjects recruited in the study were interviewed before discharge during from May 1992 to April 1993. Pregnancy and reproductive outcomes were analyzed according to their occupational classifications, adjusted for the potential confounding factors, such as age, smoking and drinking during pregnancy and history of spontaneous abortion. RESULTS: Incidence rates of pregnancy induced hypertention, low birth weight, intrauterine growth retardation and postpartum hemorrhage were higher in the workers than that of the average levels. Incidence rates of threatened abortion, gestational anemia, stillbirth and uterine atony in professional persons, of prolonged pregnancy and neonatal asphyxia in farmers, of low birth weight in housewives, of gestational anemia and of uterine atony in administrative persons all were higher than the average levels. CONCLUSION: Occupation undertaken by pregnant women was associated with the adverse pregnancy and reproductive outcomes to certain extent. PMID- 11864453 TI - [Expression and stability of fragment of Plasmodium merozoite major surface protein 1 in recombinant attenuated Salmonella typhimurium]. AB - OBJECTIVE: To determine the invasive ability of recombinant attenuated Salmonella typhimurium X4064 (pQEM1) strain containing gene fragment of Plasmodium falciparium merozoite surface protein 1 (MSP1), the stability of its plasmid, and its re-expression. METHODS: BALB/c mice were fed with recombinant attenuated S. typhimurium containing No. 1 gene fragment of P. falciparium MSP1 by gastric tube. Plate incubation, plasmid endonuclease analysis and Western blot were used to identify the recombinant attenuated S. typhimurium strains isolated from mice and the ability of re-expression, and its growth were determined in vitro. RESULTS: The attenuated strain X4064 of S. typhimurium isolated from mice contained recombinant plasmid pQEM1, no. 1 MSP1 fragment of P. falciparium was expressed in vitro in S. typhimurium X4064 (pQEM1) strain, and its growth curve of X4064 (pQEM1) strain in mice was basically similar to that of X4064. CONCLUSION: The recombinant plasmid pQEM1 could steadily exist in the X4064 strain of S. typhimurium, without influence on its invasion into host cells. X4060 (pQEM1) strain isolated from infected mice still had the ability to re express M1 protein. PMID- 11864454 TI - [Establishment of a stochastic model for variation in fecal Schistosoma japonicum egg count and estimation of its parameters]. AB - OBJECTIVE: To establish a stochastic model suitable for interpreting the variation in fecal Schistosoma japonicum egg count, and to estimate its parameters and to describe it. METHODS: Total variation in fecal egg count with a stochastic model can be divided into two kinds of sources, (1) inter-individual variation, and (2) intra-individual variation. Parameters in the model were estimated with the data collected in actual fecal egg count. RESULTS: Parameters M and r differed and k kept nearly the same in various age groups. The best cut off point of r and M for age-splitting was at eight and 12 years, respectively. Parameter k was assumed the same in different age groups in the model E, and r differed in the groups of 2 - 7 years and 8 years of age, and M differed in the groups of 2 - 11 years and 12 years of age, with a minimum value of Akaike information criterion (AIC). CONCLUSION: Age may be an important factor contributing to the estimations of parameters M and r, and the model E was the best one. PMID- 11864455 TI - [A study on the relationship between lymphocyte micronucleus rates and blood plasma benzo(a)pyrene levels in coking workers]. AB - OBJECTIVE: To study the relationship between lymphocyte micronucleus rates and blood plasma levels of benzo(a)pyrene [B(a)P] in coking workers. METHODS: One hundred coking workers and 100 unexposed workers were selected, and their lymphocyte micronucleus rates and plasma levels, as well as workplace air concentrations, of B(a)P were determined. RESULTS: It showed that there was significant difference in plasma B(a)P level between coking workers and unexposed workers, and it correlated with air concentration of B(a)P in the workplace. Positive rates of micronucleus and rates of micronucleus lymphocyte were 60% and 1.79per thousand in coking workers and 29% and 1.10per thousand in controls, respectively, with very significantly statistical difference. It also showed that positive micronucleus rate correlated with plasma level of B(a)P in dose-effect pattern. Multivariate analysis showed that factors contributing to positive micronucleus rate ranked in such an order: plasma level of B(a)P, economic status, regularity of their lifestyle and smoking status of the coking workers. CONCLUSION: Accumulation of B(a)P in blood plasma played a major role in the formation of lymphocyte micronucleus. Results of micronucleus test in coking workers could reflect preliminary damage to their health caused by it. PMID- 11864456 TI - [DNA damage in human peripheral blood lymphocyte caused by nickel and cadmium]. AB - OBJECTIVE: To understand the different implication of various forms of DNA damage in genotoxicity of nickel and cadmium. METHODS: Human peripheral lymphocyte was exposed to nickel chloride and cadmium chloride in vitro. Levels of DNA single and double-strand breaks and DNA-protein crosslinks in human peripheral lymphocyte were determined with single cell gel electrophoresis (SCGE). Activity of poly (ADP-ribose) polymerase (PARP) was determined by [(3)H]-NAD incorporating method. RESULTS: Levels of DNA single-and double-strand breaks and DNA-protein crosslinks in human peripheral lymphocyte treated with nickel and cadmium were significantly higher than those untreated, but dose-response relationship only showed in those treated with 0.10 - 10.00 micromol/L of nickel chloride and 0.16 20.00 micromol/L of cadmium. Low levels of the two kinds of metal (0.10 - 0.40 micromol/L of nickel and 0.16 micromol/L of cadmium) could induce the cleavage of DNA and activate PARP, and high levels of the two kinds of metal (2.00 - 10.00 micromol/L of nickel and 0.80 - 20.00 micromol/L of cadmium) could not induce the enzyme cleavage of DNA. CONCLUSION: Formation and cleavage of DNA double strand and blockage of activation of PARP can play an important role in carcinogenesis and mutagenesis. PMID- 11864457 TI - [Genetic polymorphism and susceptibility to occupational chronic manganism: a case-control study]. AB - OBJECTIVE: In a case-control study, the possible genetic factors relevance to occupational chronic manganism were investigated. METHODS: Forty-nine manganisms who were welders and ferromanganese smelters occupationally exposed to manganese dust and fume from three metallurgical industries, and fifty unrelated healthy control subjects who were working same workshops were recruited. The subjects were matched for sex, age, cigarette and alcohol intake. The manganese exposure duration was also matched in this case-control study. Genetic polymorphism of cytochrome P450 2D6L (CYP2D6L) gene and NAD(P)H: quinone oxidreductase (NQO1) genes from all subjects were investigated. The mutations of CYP2D6L gene located exon 6 were analyzed by a polymerase chain reaction (PCR)-based DNA amplification combined with Hha I restriction fragment length polymorphism (RFLP). The substitution of 609(C-T) at exon 6 of DT-diaphroase gene was analyzed by PCR combined Hinf I RFLP. RESULTS: The frequency of polymorphic allels, a mutation of CYP2D6, was significantly lower in patients with manganism (16.3%) than that of the controls (29.0%). A significant association was also found between the homozygote variant of CYP2D6L gene and occupational manganism. These results suggest that the CYP2D6 gene might be one of the susceptibility genes for Mn induced neurotoxicity. The allele and genotype frequencies of NQO1 gene were similar in the manganism cases and control subjects. CONCLUSION: It is possible that CYP2D6 gene may be a valuable susceptibility biomarker responded to Mn induced central nervous system disorders in workers exposed to manganese. PMID- 11864458 TI - [Effects of methylmercury on embryonic cell behavior and expression of related gene]. AB - OBJECTIVE: To explore the mechanism of embryonic developmental toxicity of methyl mercury at cellular and gene levels. METHODS: Effects of methylmercury on embryonic cell behavior and gene expression in rats were observed with in vitro (0, 0.05, 0.10, 0.20, 0.40, 0.80 and 1.60 mg/L of methylmercury) and in vivo (0, 0.2, 0.4, 0.8, 1.6 and 3.2 mg/L of methylmercury) rat models, in situ hybridization and TdT-induced dUTP nick end labeling techniques. RESULTS: Methylmercury could pass through yolk-sac placenta quickly, and inhibit placenta development and blood vessel differentiation there at higher concentration. There was a dose-response relationship between concentration of methylmercury and its embryonic developmental toxicity. Its developmental toxicity mainly characterized as patent neural tube and anomalous flexion. Methylmercury could induce excessive apoptosis of embryonic cells, inhibit apparently the synthesis of cellular DNA and RNA and damage its cellular ultrastructure. It also could induce significantly the expression of heat shock protein 70 mRNA and inhibit the expression of fibronectin and p16 mRNA. There was relationship between heat shock protein 70 mRNA, Ca(2+), apoptosis and teratogenesis. CONCLUSION: Disturbance of embryonic cell behavior and related gene expression played important roles in developmental toxicity caused by methylmercury. PMID- 11864459 TI - [Effects of methyl tertiary butyl ether on liver tissue gene expression in rats]. AB - OBJECTIVE: Methyl tertiary butyl ether (MTBE) is a new gasoline additive, with certain carcinogenecity in animal experiments. To study its possible mechanism of carcinogenesis in animals, expression of protooncogene c-myc and functional gene glutathione S-transferase-P (GST-P) in the liver tissues of rats chronically exposed to MTBE was detected. METHODS: Forty male SD rats with body weights of 180 - 200 grams were randomly divided into four groups and gavaged with MTBE dissolved in soybean oil with doses of 1 000, 600, 200 and 0 mg/kg, respectively, once daily and five days a week for 13 weeks. Liver tissues of the animals were frozen quickly in liquid nitrogen and their total RNA extracted with one-step method. The probes of c-myc and GST-P were labeled with digoxin by random primer method, and dots hybridization with RNA was used. RESULTS: Levels of expression of c-myc gene in the liver tissues of rats increased significantly, but not for those of GST-P. CONCLUSION: MTBE can induce the higher expression of c-myc gene, which suggests it can promote cell proliferation-one of possible mechanisms of carcinogenesis in animals. PMID- 11864460 TI - [Distribution of genotypes of alcohol dehydrogenase 2 and aldehyde dehydrogenase 2 in Japanese twin children]. AB - OBJECTIVE: In order to prevent alcohol related deseases, this study investigated the distribution of the genes controlling alcohol metabolism in Japan's twin. METHODS: Restriction fragment length polymorphism-polymerase chain reaction (RFLP PCR) technique was used to measure the control gene of alcohol metabolized enzymes and the genotypes of alcohol dehydrogenase 2 (ADH2) and aldehyde dehydrogenase 2 (ALDH2), which were distributed in Japan's twins. At the same time, according to the difference in genotypes, the sensitive individuals were screened from the study subjects. RESULTS: The distribution of ADH2 and ALDH2 genes were consistent with the Hardy-weinberg equation. The three genotypes of ADH2 gene were ADH2(1)/ADH2(1) (1.1%), ADH2(1)/ADH2(2) (44.6%) and ADH2(2)/ADH2(2) (54.3%). And those of ALDH2 gene were ALDH2(1)/ALDH2(1) (41.3%), ALDH2(1)/ALDH2(2) (39.1%) and ALDH2(2)/ALDH2(2) (19.6%). The frequency of ADH2 and ALDH2 genes was 0.255, 0.745 and 0.609, 0.391 respectively. CONCLUSION: Not only the distribution of genotypes of ADH2 and ALDH2 is known, but also the sensitive individuals are found, which can help prevent alcohol related disease. PMID- 11864461 TI - [Relationship between obese gene expressive product and simple obesity in children]. AB - OBJECTIVE: To investigate the relationship between obese gene expressive product obese protein (OP) and the pathogenesis and prognosis of simple obesity in 30 children with normal body weight (control group). METHODS: Before immunoassay, OP was extracted from the whole blood samples with Sep-pak C(18) Cartridge. RESULTS: The plasma contents of OP in the obese group were (118.53 +/- 25.02) ng/L and (197.14 +/- 26.83) ng/L (P < 0.001), respectively. In the obese group, plasma OP was not detected in three cases and was (19.90 +/- 3.94) ng/L in other three cases. These obese children with severe OP deficit not only had more increased body mass index, and levels of total cholesterol, triglyceride and low-density lipoprotein-cholesterol, but also had lower therapeutic effect (16.67%) than those with mild OP depletion (79.16%). CONCLUSION: The children with simple obesity have OP deficit. Simple obesity may be related with the reduced plasma content of OP. The obese children with severe OP deficit have poorer curative effect and unfavorable prognosis. PMID- 11864462 TI - [Protection against synergistic hepatocarcinogenesis of hepatitis B virus expression and aflatoxin B1 by antioxidant 2(3)-tert-4 -hydroxyanisole (BHA) in HBV large envelope transgenic mice]. AB - OBJECTIVE: To investigate the effects of 2(3)-tert-4-hydroxyanisole (BHA) on protection against synergistic hepatocarcinogenesis of aflatoxin B(1) (AFB(1)) and hepatitis B virus (HBV) expression in HBV large envelope transgenic mice. METHODS: Protective effects of dietary antioxidant BHA on life span was observed in 49 cases of HBV transgenic mice and 48 cases of non-transgenic mice by determinations of enzyme activities and liver MDA, and by direct detection of liver oxidative free radicals (OFR) using electron spin resonance (ESR). RESULTS: In the HBV transgenic mice which exposed to AFB1, the incidence of hepatocellular adenoma was 17% (2/12), but no carcinoma was found in BHA group. In the regular diet group, that was greater with 67% (6/9) of adenoma and 22% (2/9) of carcinoma. BHA could decrease significantly the concentrations of liver MDA and OFR, compared with those with regular diet. The activities of quinone reductase and glutathione S-transferase in liver cytosols increased by 3 - 7 times, as in the controls, in response to BHA. CONCLUSIONS: Addition of BHA to the diet resulted in significantly elevation of phase II enzyme activities in liver. BHA could directly eliminate liver OFR and inhibit growth of hepatocellular altered foci. These actions may effectively put off hepatocellular carcinogenesis in mice. PMID- 11864463 TI - [Detection and proliferation of hemorrhagic fever virus in chigger mites]. AB - OBJECTIVE: To study the impact of Leptotromhidium scutellaris as a vector in transmission of hemorrhagic fever. METHODS: Reverse transcription-polymerase chain reaction (PCR) and in situ hybridization technique were used to detect RNA of hemorrhagic fever with renal syndrome virus (HFRSV). TCID(50)/ml of HFRSV were titrated for larva and nymph of chigger mites periodically. RESULTS: RNA of HFRSV was detected in the tissues of chigger mites, such as ovary cells, etc. by PCR and in situ hybridization. Titration of TCID(50)/ml showed that HFRSV could be transmitted transstadially and proliferated in chigger mites. CONCLUSION: It provides direct evidence at molecular level for the role of chigger mite as a vector in transmission of HFRSV with theoretical and practical importance in prevention of HFRS. PMID- 11864464 TI - [Synthetic assessment of health impact of cerebrovascular diseases on potential health days of life lost]. AB - OBJECTIVE: To assess synthetically the impact on population health caused by cerebrovascular diseases. METHODS: Incidence, duration of disease, disabilities and deaths of cerebrovascular diseases were analyzed with potential health days of life lost (PHDLL) as an indicator in urban areas of Changsha, Hunan Province. RESULTS: The PHDLL caused by cerebrovascular disease totaled 2 624.94 days per thousand of population, 33.01% of them attributed to deaths and 53.44% to chronic disability. CONCLUSION: Hemorrhagic cerebrovascular disease mainly lead to death and ischemic one caused chronic disability. PMID- 11864465 TI - [Studies on immunization strategies for hepatitis B in different endemic areas of China]. AB - OBJECTIVE: To compare the cost-benefit of immunization with hepatitis B (HB) vaccine in different endemic areas of HB and to explore the optimal strategy for it. METHODS: Cost-benefit analysis and comprehensive weighted score analysis (CWSA) were used to screen the optimal immunization strategy for low, medium and heavy endemic areas of HB. RESULTS: Significant economic benefit was obtained in Long-An County, Guangxi Zhuang Autonomous Region, Shanghai and Jinan, Shandong Province from HB immunization programs for infants. The greatest benefit cost ratio (BCR) was provided by the low-dose immunization strategy, which was 49.91, 54.53 and 37.68, respectively for heavy, medium and low endemic areas of HB. The maximal net benefit could be obtained by the high-dose immunization strategy with better immune protective efficacy. CWSA showed that the low-dose immunization strategy was the optimal one for HB immunization. CONCLUSION: It is suggested that the low-dose immunization strategy be implemented in undeveloped areas to obtain a greater BCR, and the high-dose one be implemented in more developed areas to obtain more net benefit and lower prevalence of hepatitis B surface antigen carriage. PMID- 11864466 TI - [Energy dispersive X-ray analysis in studying the permeability of blood-brain barrier caused by lead in rats]. AB - OBJECTIVE: To observe the changes in permeability of blood-brain barrier caused by lead in rats. METHODS: Rats were fed with drinking water containing 10 and 30 mg/L of lead for three months. Electron microscopy lanthanum nitrate-tracing and electron probe-energy dispersive X-ray (EDX) analysis were used to observe the distribution of lead in different cells of bilateral sides of capillaries of cerebral cortex and the changes in their permeability. RESULTS: There was significant difference in blood lead levels between the rats with drinking water of 10 mg/L lead and the controls, but only a peak of lead, L(alpha), could appear in plasmalemma of endotheliaocyte endosurface in capillaries, and no any peak in the basement membrane of endotheliocyte of the outer-side, end feet of astrocytes and axons of neurons. Lanthanum nitrate-tracing showed changes in capillary permeability. But, in those with drinking water of 30 mg/L of lead, peak of lead, L(alpha) and M(alpha) could appear in all cells mentioned above. Meanwhile, lanthanum salt granules, which penetrated the cellular gaps of endothelia and seeped in the basement and its peripheral cells, could be found in certain sites of the capillaries with larger diameters. CONCLUSION: It suggests that the plasmalemma of endotheliocyte endosurface was the initial target in brain, where was affected by lead, and the consecutive endotheliocytes of blood vessels served as a blood-brain barrier to lead, although in a very weak action. A little bit accumulation of lead on the plasmalemma of endotheliocyte endosurface could cause damage in the close junction between endotheliocytes to spread to brain substance, same as the lanthanum tracing showed. PMID- 11864468 TI - [Effects of homocysteine on cardiovascular development in early chicken embryo]. AB - OBJECTIVE: To study the relationship between homocysteine (HCY) and cardiovascular development in the early embryo and to determine whether folic acid and cobalamine (Vit B(12)) can reduce the developmental toxicity of HCY to cardiovascular system. METHODS: Incubation test for chicken embryo, electron microscopy, methylgreen-pyronine stain, in situ fragment end labeling, and intervention trial with folic acid and vitamin B(12) were used in the study. RESULTS: Treatment of 0 - 16.0 micromol of HCY/per embryo could disturb their heart development and differentiation of blood vessels at fetal ages of two and four days, in a dose-response pattern, with 24.1% and 25.0% of heart defect and 60.7% of inhibition of blood vessels of yolk sac in 8.0 micromol of HCY, respectively. A dose of more than four micromol/per embryo could damage the structure of myocardial cells and organelle, and inhibit the synthesis of cellular DNA and RNA. It was the first time to find that 8.0 micromol of HCY could induce excessive apoptosis of myocardial cells (2.7%), significantly higher than that in the normal control group. Folic acid could antagonize the cardiovascular toxicity caused by HCY, stronger than what Vit B(12) could do. CONCLUSION: HCY was a new risk factor which could damage cardiovascular development, and folic acid could effectively antagonize its developmental toxicity. Cellular apoptosis might relate to cardiovascular defect. PMID- 11864467 TI - [Application of cerium cytochemical method in observation of renal injury caused by oxygen free radical with cadmium induction]. AB - OBJECTIVE: To study acute renal injury caused by oxygen free radical with cadmium induction. METHODS: Electron microscopy cytochemical and energy dispersive X-ray (EDX) analysis techniques were used. RESULTS: Cadmium ions could enter the epithelial cells of renal proximal convoluted tubules. Excessive production of oxygen free radical was found in the kidney tissues, which located mainly in the endothelial cells of blood vessels, mitochondria of epithelial cells of renal proximal convoluted tubules, microvilli and basement membrane. Swelling of epithelial cells and mitochondria and drop of epithelial cells could occur in renal proximal convoluted tubules. CONCLUSION: It suggests that cadmium ions can induce the formation of large amount of oxygen free radical and cause renal injury. PMID- 11864469 TI - [Effects of occupational exposure to formamidines on cardiovascular functions]. AB - OBJECTIVE: To understand the possible effects of occupational exposure to formamidines on human cardiovascular function, this study was carried out. METHODS: Sixteen farmers spraying chlordimeform, 14 packers packaging chlordimeform and 23 packers packaging mono-formamidine were followed-up pre- and post-exposure. Their urinary excretion of formamidine or its metabolite, as well as air concentrations of formamidine at their work places and their skin contamination with it were measured to estimate the exposure level. Furthermore, 24-hour urinary level of vanillinmandelic acid (VMA) was analyzed among mono formamidine packers. RESULTS: Their whole body skin contaminated with 3.240 and 2.142 g/cm(2) of chlordimeform in the sprayers and packers, respectively. Their hand skin contaminated with mono-formamidine at 6.59 g/cm(2) in the packers. It indicated that the major route of exposure to formamidine was skin contamination. Urinary levels of formamidines or their metabolites increased significantly after exposure, reaching 6.194 and 3.378 micromol/L for the sprayers and packers exposed to chlordimeform, respectively, and 2.760 - 3.427 micromol/L for mono formamidine in the packers. Their heart rates slowed down, P-R and Q-T intervals prolonged and blood pressure reduced after exposure, as compared with those before exposure. Consistency of the results in several studies demonstrated that formamidines could induce changes in the indices for cardiovascular functions under the relatively low exposure levels at present. Decrease of urinary VMA from 0.068 micromol/L pre-exposure to 0.040 - 0.055 micromol/L post-exposure suggested that catecholamine could play a role in these effects. CONCLUSION: Formamidines has certain effects on human cardiovascular function. It is suggested that attention be paid to the changes in cardiovascular functions of those exposed in their health surveillance. PMID- 11864470 TI - [Quality of life as a time-dependent covariable in cox regression analysis and its application]. AB - OBJECTIVE: To present a statistical method for combined evaluation of the quality of life and survival time and to study some issues in its application. METHODS: Based on Cox regression model, quality of life was used as a time-dependent covariable in analysis of the influence factors for the effectiveness of detoxification in 212 drug addicts. RESULTS: The influence factors entered the regression equation were ways of detoxification for drug abusers, marital status, mode of drug abuse, frequency of detoxification, maternal educational level, maternal occupation, family atmosphere and help and encourage from their relatives and friends, with their hazard ratios of 0.673, 1.315, 1.319, 0.867, 1.120, 1.318, 0.724 and 1.150, respectively. CONCLUSION: Combined statistical analysis of quality of life and survival time was a better method for the evaluation of the effectiveness of detoxification and its influence factors, which depended mainly on their mother's qualification, family atmosphere, mode and length of drug abuse, etc. PMID- 11864471 TI - [Nucleotide sequence analysis for high variance region of hepatitis C virus in patients with hemodialysis]. AB - OBJECTIVE: To investigate the route of transmission for hepatitis C virus (HCV) infection in patients on hemodialysis (HD). METHODS: Nucleotide sequence of high variance region (HVR) of HCV was analyzed with nucleotide sequencing technique in 15 HD patients, and its homology was compared. RESULTS: Comparison of the nucleotide sequences of HCV HVR in 15 HD patients showed that their homologies in isolates 8, 12 and 14 reached 97.50%, 95.00% in isolates 4, 5 and 10, and more than 80.00% in isolates 1, 3 - 8 and 10 - 14. Clinical data showed that all these patients were hemodialyzed in the same room with the same HD machine. Isolates 2 and all others were 61.25% - 66.25% in homology, and 57.50% - 67.50% in isolates 9 and all others, and both the patients had histories of large-quantity blood transfusion. CONCLUSION: Blood transfusion was the primary risk factor for HCV infection in HD patients, but iatrogenic transmission of HCV through HD environment could also exist. PMID- 11864472 TI - [Evaluation of potential harmful effects of organic pollutants in tap water on mice in vivo]. AB - OBJECTIVE: To study the harmful effects of organic concentrates from tap water on mammal animals in vivo. METHODS: Organic pollutants were absorbed with resin XAD 2 from tap water and administered the mice via a stomach tube or intraperitoneal injection. Micronucleus rate in bone marrow polychromatic erythrocytes, testicular chromosome aberration rate, indices for immune functions and the activity of ethoxyresourufin o-deethylase (EROD) in S(9) fraction of the liver were determined. RESULTS: Micronucleus rate and chromosome aberration rate increased by 1.7 and 3.5 folds, respectively, after administration of high dose of organic pollutants from the tap water (200 liters of tap water per kilograms of body weight) for three days. Humoral immunity and non-specific immune functions were obviously inhibited after administration of low-dose (6 L/kg). In addition, organic concentrates from tap water also could significantly induce the activity of EROD, with a good dose-dependent relationship. CONCLUSION: Organic concentrates from tap water could cause chromosome damage to somatic and germ cells and decrease in immune functions in the mice, as well as they could induce the activity of cytochrome P448 in the liver microsome. It suggested that organic pollutants concentrated from tap water had potential carcinogenic effects on mammal animals. PMID- 11864474 TI - [Component in diphtheria-pertussis-tetanus-hepatitis B vaccine]. AB - OBJECTIVE: To study immunogenecity of yeast-derived recombinant hepatitis B (YHB) component in diphtheria-pertussis-tetanus-YHB vaccine (DPTw-YHB). METHODS: Immunogenecity of tetra-valent DTPw-YHB vaccine and mono-valent recombinant YHB vaccine, and that of the tetra-valent vaccine with varied YHB component were compared. The efficiency and stability of recombinant YHB in the tetra-valent vaccine stored at 2 - 8 degrees C for 18 months were determined. RESULTS: The efficiency of recombinant YHB in the tetra-valent vaccine enhanced significantly in mice, as compared with that of mono-valent recombinant YHB vaccine, with an average mouse ED(50) of 1:2.0 - 1:3.1. There was no significant difference in efficiency of the tetra-valent vaccine with varied recombinant YHB component. Recombinant YHBin the DPTw-YHB tetra-valent vaccine still kept good stability stored at 2 - 8 degrees C for 18 months. CONCLUSION: Recombinant YHB in the tetra valent vaccine was more immunogenic than the mono-valent YHB vaccine. No interference and inhibition of DPTw to recombinant YHB was found, indicating good compatibility between DPTw and YHB. PMID- 11864473 TI - [Studies on apoptosis of human lung adenocarcinoma cells induced by oltipraz]. AB - OBJECTIVE: To study whether oltipraz can induce apoptosis of human lung adenocarcinoma (GLC-82) cells and its possible mechanism. METHODS: Light microscopy, DNA electrophoresis, flow cytometry and video time-lapse monitoring were used to observe apoptosis of GLC-82 induced by oltipraz. RESULTS: GLC-82 cells treated with oltipraz underwent the arrest of cell mitosis at metaphase, higher percentage of G(2)/M and peak of apoptosis were found, and cell body shrunk, nuclear chromosome condensed or fragmented and nuclear DNA fragment displayed "laden" bands in apoptic cells. Apoptosis of GLC-82 induced by oltipraz was more obvious at a concentration of 120 microg/ml. CONCLUSION: Oltipraz could induce apoptosis of GLC-82 at certain concentrations, which associated closely with the arrest of mitotic cycle. PMID- 11864475 TI - [A prevalence study on hepatitis C infection in 4,055 healthy children of Beijing]. AB - OBJECTIVE: To understand the prevalence of hepatitis C (HC) infection in children of urban Beijing. METHODS: Serum antibodies against hepatitis C virus (anti-HCV) and HCV RNA were determined with enzyme-linked immunosorbent assay (ELISA) in 4 055 healthy children under 14 years of age living in four urban districts of Beijing. RESULTS: Fourteen children were positive for anti-HCV, and four of them were positive for HCV RNA as well, with a prevalence of HCV infection of 0.35%. There was no significant difference in prevalence of HCV infection between children living in different districts and by sex and age. However, very significant difference in prevalence of HCV infection between children with history of blood transfusion and injection of human blood-derived globulin and those without it. The parents of 14 children with positive anti-HCV were all negative for it. CONCLUSION: Blood-borne transmission still was the main route for HCV infection in children. PMID- 11864476 TI - [Studies on appropriate methods for dust scrubbing from dust-laden gas in shaft kilns of small-sized cement plants]. AB - OBJECTIVE: To study an appropriate method for dust scrubbing from dust-laden gas in the shaft kilns of small-sized cement plants. METHODS: Characteristics of dust laden gas, including its temperature, dew point, dust concentration, diameters of dust particles and their distribution, as well as fractional dust collection efficiencies of the dust precipitator settling chamber, cyclone, impinging scrubber, impinging-spray scrubber, were measured according to the National Standards for Measurement of Particulate in Exhaust Gas Emitted from the Stationary Pollution Sources and Sampling Methods of Gaseous Pollutants (GB/T16157 - 1996), with a WY-1 In-stack 7-Stage Cascade Impactor for fractional dust collection. RESULTS: The fractional dust collection efficiency of the settling chamber for the particles greater than 12 microm was 72%, while that of the cyclone for the particles of 2 microm, 5 microm and 10 microm were 40.0%, 72.5% and 88.9%, respectively, and that of impinging scrubber was 55.0%, 91.0% and 98.9%, respectively; and that of impinging-spray scrubber were 96.0%, 99.2% and 99.6%, respectively. Results showed that 81% of the dust (by weight) from the shaft kilns were greater than 10 microm, so the settling chamber was effective for the removal of this fraction of dust. But dust particles behind the settling chamber were finer, and about 59% - 69% of them were less than 2 microm in diameters. In that case, the cyclone, impinging scrubber and impinging-spray scrubber could give an overall collection efficiency of 54%, 57% and 96%, respectively. Based on this, the following combination of dust collectors were adopted to meet the national standards for exhaust emission: a settling chamber plus an electrostatic precipitator, a settling chamber plus an impinging-spray scrubber and a settling chamber plus an impinging scrubber. The cement plants could select appropriate dust collectors according to their technical and economical conditions. CONCLUSION: Based on the results of the studies, appropriate dust collectors used to treat dust-laden gas from shaft kilns to meet the national standards for exhaust emission were recommended. PMID- 11864477 TI - [Changes in expression of genes for insulin, glucagon and IGF-II in neonatal rats with malnutrition]. AB - OBJECTIVE: To study the effects of malnutrition on the function of cells in fetal islet of pancreas. METHODS: A neonatal rat model with malnutrition was prepared by maternal food restriction (fed with less than half amount of the normal) during the 14th to 21st days of pregnancy. Expression of genes for insulin and glucagon in the pancreas, as well as expression of genes for insulin-like growth factor II (IGF-II) in the pancreas and liver of the rats were analyzed with Northern blotting technique. RESULTS: Pancreatic content of insulin mRNA was significantly lower in neonatal rats with malnutrition than that in the normal group (P < 0.05), but no obvious changes in glucagon mRNA in the pancreas was found. There was no significant difference in IGF-II mRNA content in the pancreas and liver between the neonatal rats with malnutrition and the normal control group. CONCLUSION: Malnutrition during rat fetal development could cause inhibition of expression of insulin gene, but no obvious effects on the expression of glucagon and IGF-II genes. PMID- 11864478 TI - [A study on family aggregation of type 2 diabetes mellitus]. AB - OBJECTIVE: To estimate the genetic susceptibility and its relative risk (RR) in the first degree relatives of type 2 diabetes mellitus (DM) through family aggregation analysis. METHODS: A population-based case-control study including 654 pedigrees (363 index cases and 291 controls) was carried out. RESULTS: The prevalence rate in the first degree relatives of DM probands was 3.94%, significantly higher than that in the controls (1.09%), with a relative risk of 3.62 (chi(2) = 36.5, P < 0.001). The RRs in different consanguineal relatives were all higher than 3.0. The younger was the probands at diagnosis for DM, the higher its prevalence rates and RRs among their relatives was and the more likelihood that more than one case of DM occurred in them. CONCLUSION: Significant family aggregation and higher genetic susceptibility were found in the first degree relatives of DM, on whom prevention and treatment for DM should be focused. PMID- 11864479 TI - [Association of body size at birth with impaired glucose tolerance during their adulthood for men and women aged 41 to 47 years in Beijing of China]. AB - OBJECTIVE: To explore if there was relationship between small body size at birth and type 2 diabetes and impaired glucose tolerance (IGT) during adulthood in China. METHODS: Six hundred and twenty-eight singletons born in the Peking Union Medical College Hospital in Beijing during July 1948 to the end of 1954 were followed-up and their medical records at birth were abstracted. Anthropometry and standard oral glucose tolerance test were carried out for all of them. Plasma insulin level was measured with radioimmunoassay for them. RESULTS: Prevalence of type 2 diabetes and IGT decreased with the increase in their birth weight, Ponderal index (PI) and head circumference at birth (for trend test, chi(2) = 6.7, P = 0.01; chi(2) = 4.8, P = 0.03; and chi(2) = 5.8, P = 0.02; respectively), with the highest of 43.8% in those with thin body size (PI < 24 kg/m(3)) at birth and obesity (BMI < 75 percentile) during adulthood, and the lowest of 8.3% in those with more fat at birth (PI >or= 28 kg/m(3)) and keeping relatively thin (BMI < 25th percentile) during adulthood. BMI of mothers in their early and late pregnancy correlated reversely with blood glucose levels two hours after sugar load for their children in adulthood. In addition, body weight, PI and head circumference at birth correlated reversely with their plasma insulin levels fasting and two hours after sugar load and levels of 32 -- 33 split pro-insulin during their adulthood. After adjustment for current BMI, lifestyle and economic status, body size at birth mentioned above still associated with their glucose tolerance and level of plasma insulin during adulthood. CONCLUSION: There was relationship between small body size at birth and prevalence of type 2 diabetes and IGT during adulthood. Prevalence of type 2 diabetes and IGT depended on the synergic effect of thin body size at birth and obesity during adulthood. PMID- 11864480 TI - [Effect of fodder components on the product of obese gene expression--leptin in rats]. AB - OBJECTIVE: To study the effects of different fodder components and levels of caloric uptake on the product of obese gene expression -- leptin and in vivo secretion of insulin and growth hormone, and to study the relationship between them in rats. METHODS: Plasma leptin was determined by Western blot, and serum insulin and growth hormone were determined by radioimmunoassay (RIA). RESULTS: There was no significant effects of fodder components, including high protein, fat and carbohydrate, on the levels of leptin, insulin and growth hormone in rats, with the same caloric uptake in them. However, with the higher caloric uptake, level of obese gene expression increased, showing increased levels of plasma leptin and serum insulin and decreased level of serum growth hormone. CONCLUSION: There could existed an interaction between expression of obese gene and secretions of leptin, insulin and growth hormone, which could be modified by the same metabolic signal. PMID- 11864481 TI - [Studies on changes in levels of hormone and expression of ob gene during induction of obesity in rats]. AB - OBJECTIVE: To study the rules of changes in ob mRNA and its relation to hormone level in vivo in rats during their dynamic process to obesity induced by high caloric fodder. METHODS: Level of ob mRNA in adipose tissues of rats was determined with dot-hybridization techniques, and levels of serum insulin and growth hormone were measured with radioimmunoassay. RESULTS: Level of ob mRNA in the fatty tissues and serum insulin level increased, but serum level of growth hormone decreased, with the increase in body weight of rats. CONCLUSION: Increase in expression of ob mRNA could be the result, but not the cause, of obesity. Changes in the levels of ob mRNA, serum insulin and growth hormone indicated that they could affect each other and could be regulated by common metabolic signals. PMID- 11864482 TI - [Zinc promotes proliferation and differentiation of osteoblast in rats in vitro]. AB - OBJECTIVE: Effect of zinc on proliferation and differentiation of osteoblast in rats was studied in vitro. METHODS: Osteoblasts were isolated from the rat calvaria and subcultured in DMEM media. Rats were divided into four groups in the study, with administration of 0 micromol/L (as control), 10 micromol/L, 25 micromol/L and 50 micromol/L of Zn(2+), respectively. DNA synthesis at varied time points was determined with incorporation of (3)H-thymidine, and cell cycle was analyzed by flow cytometry. Synthesis of collagen was measured with incorporation of (3)H-proline. Activity of alkaline phosphatase (AKP) and osteocalcin content were measured by enzyme dynamics method and radioimmunoassay, respectively. RESULTS: Incorporation of (3)H-thymidine in the three groups with different zinc-supplement was significantly higher than that in the control group at each time point, and supplement with 25 micromol/L and 50 micromol/L of Zn(2+) could promote the transition of cell cycle from phases G(0)/G(1) to S phase. Syntheses of collagen and osteocalcin and activity of AKP in the groups with zinc supplement were greater, as compared with those in the control group. CONCLUSION: Zinc could promote proliferation and differentiation of the rat osteoblast in vitro. PMID- 11864483 TI - [Time trend of mortality rates of cerebrovascular diseases in Hong Kong]. AB - OBJECTIVE: To describe the time trend of mortality rates of cerebrovascular diseases (CVD) in Hong Kong for the period of 1976 to 1995, and to compare them with those in the more developed countries around the world. METHODS: Mortality rates of CVD were standardized directly using world standard population in 1961. Annual percent changes of mortality rates of CVD were estimated with a non-linear regression. Birth-cohort analysis was carried out at an interval of five-year of age to compare their age-specific mortality rates of CVD in different birth cohorts. Time trend of cause-specific mortality rates of CVD both in males and females was fitted with a simple linear regression model. RESULTS: Considerable downward trend of CVD mortality was observed during the past twenty years. Mortality rates among men and women decreased by 50.96% and 37.85% in 1995, as compared to those in 1976, respectively. Although in general, a greater trend in CVD mortality was observed during the last 10 years, as compared with that in the previous 10 years, an annual increase in CVD mortality of 1.34% was found among males aged 35 to 44 years during the latter 10 years. CVD mortality for both males and females decreased steadily by 1% per year. CVD mortality for females will catch up with that for males by the year of 2013, if such a trend continues. Birth-cohort analysis showed that those born more recently at the same age-group had lower mortality of CVD (except for those born after 1955). Data from hospital admission showed that improvement in the treatment for CVD could have contributed partly to the decrease in its mortality. CONCLUSION: Time trend of mortality of CVD in Hong Kong was similar to that in many other economically developed areas around the world. PMID- 11864484 TI - [A case-control study on cerebral palsy in children]. AB - OBJECTIVE: To identify the risk factors for cerebral palsy (CP) in children. METHODS: A population-based 1:2 matched case-control study was conducted during May to June 1997 in southern Jiangsu Province of China. RESULTS: Risk factors for CP in children could be summarized into four categories, i.e.; (1) fetal growth retardation, preterm delivery and low birth weight; (2) asphyxia at birth, intracranial hemorrhage, ischemic and hypoxic encephalopathy and hyperbilirubinemic encephalopathy; (3) delivery with vaccum suction and traumatic brain injury; and (4) other prenatal factors. CONCLUSION: The first and second categories of risk factors correlated strongly and reproducibly with CP in children. It is emphasized that antenatal care be enforced to prevent occurrence of risk factors, such as low birth weight, etc., and the newborns with asphyxia be vigorously rescued and put under close supervision. For risk factors in the third category, the key measures should be obstetric practice and health education for the parents. But, little about the knowledge of the risk factors in the fourth category has been understood, which could be associated with those in the first two categories. PMID- 11864485 TI - [Effects of cigarette smoke solution and chrysotile on proliferation activity of human embryo lung cells]. AB - OBJECTIVE: To study the separate and combined effects of cigarette smoke solution and chrysotile on cell proliferation activity. METHODS: Human embryo lung (HEL)fibroblasts were cultivated with cigarette smoke solution and chrysotile separately or in combination at varied concentrations for certain times, and cell proliferation was observed with [(3)H] TdR incorporation. RESULTS: Cigarette smoke solution and chrysotile could enhance cell proliferation at lower doses, both separately or in combination, but inhibit it at higher doses. Lower dose of cigarette smoke solution and chrysotile in combination could enhance cell proliferation synergistically, but higher doses could only do additively. CONCLUSION: Cigarette smoke solution and chrysotile could affect HEL cell proliferation separately or in combination, with an enhancing or inhibiting effect, which depended on their doses. PMID- 11864486 TI - [Effects of tantalum and its oxide on exposed workers]. AB - OBJECTIVE: To explore effects of tantalum and its oxide on the exposed workers. METHODS: Health examination for 113 workers exposed to tantalum and its oxide and a field survey for occupational hygiene in the workplace were carried out. RESULTS: Workplace air concentration of tantalum and its oxide averaged 0.1 - 7.6 mg/m(3). There was no significant difference in prevalence of symptoms and signs and biochemical, pulmonary function, chest X-ray and electrocardiograph examinations between the exposed and control workers. CONCLUSION: No adverse effect of tantalum and its oxide on the exposed workers was found. PMID- 11864487 TI - [The effects of lead poisoning on expression of nerve growth factor gene of submandibular gland in mice]. AB - OBJECTIVE: To study the toxicity of lead poisoning to submandibular gland and its effects on nerve growth factor (NGF) gene expression in mice. METHODS: An experimental model with lead poisoning was established and its histopathological changes in the submandibular gland of mice were observed under light and electronic microscopes. Effects of lead poisoning on expression of NGF mRNA in submandibular gland were analyzed quantitatively by human NGF DNA probe labelled with digoxin with in situ hybridization. RESULTS: Body weight of the mice with experimental lead poisoning reduced, and their lead levels in blood and submandibular gland increased. Lobular atrophy, fibrous hypertrophy, angiectasis of the stroma and enlargement of lobule interstitial in mouse submandibular gland with lead poisoning could be found under light and electron microscopes. Their rough surfaced endoplasmic reticulum was extended and mitochondrion swollen. Graphic analysis showed that diameters of the secretory striate ducts and granular tubules decreased in the mice with lead poisoning. Results of in situ hybridization indicated that hybridized signals in the granular and secretory striate ducts and granular tubules reduced significantly, and NGF mRNA expression decreased. CONCLUSION: Lead is toxic to the submandibular gland of mice and can affect their NGF gene expression. PMID- 11864489 TI - [Effects of environmental lead exposure in kindergartens on children's blood lead level]. AB - OBJECTIVE: To determine if kindergartens' environmental status can influence children's lead exposure. METHODS: Environmental specimens, such as floor dust, peeled-off paint chip, soil and drinking water, as well as children's hand dust and blood samples, were collected and measured for their lead levels in 19 kindergartens, to analyze the relationship between children's blood lead levels and their environmental lead exposure. RESULTS: Geometric means of lead concentrations in indoor floor dust, peeled-off paint chip, dust fallout, outdoor floor dust, soil and drinking water were 86.5 microg/m(2), 235.5 microg/g, 445.9 microg/g, 172.4 microg/m(2), 70.1 microg/g and 12.5 microg/L, respectively. Lead level on children's hands averaged 3.4 microg at both hands. Blood lead levels in children correlated positively with the lead concentrations of outdoor floor dust and their hand dust, with correlation coefficients of 0.5186 and 0.2206, respectively. Multiple regression analysis showed that hand dust lead level in children entered the regression model with a largest standardized partial regression coefficient of 0.3842 and a coefficient of determination of 0.673 for the full equation with F = 6.52 and P < 0.01. CONCLUSION: Status of environmental health in kindergartens plays an important role in children's lead exposure. It is necessary to offer health education for children and make them wash their hands often and overcome unhealthy behavior of sucking their fingers. PMID- 11864488 TI - [A study on relationship between blood lead level and physical growth and development of babies and young children in Shanghai]. AB - OBJECTIVE: To study the effects of low level lead exposure on physical growth and development in young children. METHODS: Blood lead level and indicators for physical growth and development, such as body height and weight, and head and chest circumferences were measured in randomly sampled 1,969 children aged 1 - 6 years in Shanghai. Z scores for these indicators were calculated for each of them based on their age and sex-specific norms for children in urban and suburban Shanghai. And, regression analysis of blood lead level was conducted on them. RESULTS: Geometric mean of blood lead level was 0.400 micromol/L in young children of Shanghai, and 744 (37.8%) of 1,969 children were equal to or higher than 0.483 micromol/L. Geometric mean of blood lead level correlated in reverse to the Z scores for their body height, weight and head circumference, even if adjusted for the potential confounding factors. CONCLUSION: Low level lead exposure could cause adverse effects on their physical growth and development in young children of Shanghai. PMID- 11864490 TI - [Re-assessment of indicators for screening lead poisoning]. AB - OBJECTIVE: To determine the accuracy of the indicators for detecting lead poisoning. METHODS: Blood and urine specimens in 157 workers exposed to lead were collected. Their blood levels of lead, free erythrocyte protoporphyrin (FEP), zinc protoporphyrin (ZPP), urine delta-aminolevulinic acid and activity of red cell pyrimidine 5'-nucleotidase were determined, with gold standards of blood lead levels of >or= 1.93 micromol/L and >or= 2.90 micromol/L. Accuracy assessment of the indicators mentioned above for detecting lead poisoning was based on the area under the receiver operative characteristic (AUC(ROC)) curve with a software specially for ROC curve analysis and software of Epi-Info, after determining the optimal cut-off points for the above mentioned indicators in detecting those with blood lead >or= 1.93 micromol/L and >or= 2.90 micromol/L. The sensitivity of screening in parallel for detecting those with blood lead >or= 1.93 micromol/L and >or= 2.90 micromol/L was calculated and compared. RESULTS: For detecting those with blood lead level >or= 1.93 micromol/L, the area under ROC curve (AUC(ROC)) of above indicators was 0.978 for ZPP, 0.973 for red cell P5'N, 0.937 for FEP, 0.890 for urine delta-ALA (significantly different from that for ZPP with P < 0.05), and 0.845 for urine lead level (significantly different from that for ZPP with P < 0.05), respectively. For detecting those with blood lead level >or= 2.90 micromol/L, the area under ROC curve (AUC(ROC)) of above indicators was 0.975 for red cell P5'N, 0.954 for ZPP, 0.906 for FEP (significantly different from that for red cell P5'N with P < 0.05), 0.900 for urine delta-ALA (significantly different from that for red cell P5'N with P < 0.05), and 0.750 for urine lead level (significantly different from that for red cell P5'N with P < 0.05). Sensitivity of screening in parallel with ZPP and P5'N was significantly higher than that with urine levels of lead and delta-ALA. CONCLUSION: Determinations of red cell P5'N and ZPP for detecting those with blood lead >or= 1.93 micromol/L and >or= 2.90 micromol/L are more accurate. PMID- 11864491 TI - [A case-control study on Guillain-Barre syndrome in children of North China]. AB - OBJECTIVE: This study aimed at exploring the risk factors for Guillain-Barre syndrome (GBS). METHODS: A case-control study design was used with 51 cases of GBS, diagnosed based on their symptoms, signs and electrophysiological examinations and exclusion of poliomyelitis and other acute flaccid paralysis, and 51 controls matched on age, sex and resident village. Serum IgG antibodies specific for Campylobacter jejuni were determined for all the subjects by enzyme linked immunosorbent assay (ELISA) with a preparation of surface antigen of C. jejuni C(1) strain isolated from the patients and prevalent in north China. Each case and control were interviewed with his/her parents or guardians by a trained interviewer using an ad hoc questionnaire, including his/her demographic information, socioeconomic status, onset of the illness, and potential risk factors in their environment and personal hygiene. Data were analyzed with SAS software release 6.04 in a microcomputer. RESULTS: GBS was associated with a few factors, such as residential areas (45 of the 51 cases living in the rural areas, accounting for 88.2% of the total), polio and hepatitis B vaccine immunization before onset of the illness (with ORs of 7.27 and 3.14, respectively), no hand washing after defecation and before meals (with an OR of 6.15) and getting cold and going to the river or lake site before onset of the illness (with ORs of 13.75 and 12.20, respectively). Infection with Campylobacter jejuni associated strongly with the illness (with an OR of 9.5, P < 0.001). Thirty-five of the 51 cases had precursor symptoms before onset of the illness (68.6%). CONCLUSION: It suggests that occurrence of GBS may correlate to infection with Campylobacter jejuni and poor personal hygiene in children. PMID- 11864492 TI - [Analysis of bone mineral density and relevant factors in 781 healthy people aged 15 to 50]. AB - OBJECTIVE: To determine further the age to form peak bone mass (PBM) and the factor influencing it and bone mineral density (BMD) and to find an effective way to prevent from osteoporosis in the elderly. METHODS: BMD was measured in 781 people aged 15 to 50 years and relevant factors in lifestyle were investigated. Multivariate regression was used to analyze the relationship between BMD and lifestyles and to identify both risk and benefit factors for osteoporosis. RESULTS: For every ten-kilogram increase in body weight, BMD at varied sites increased by an amount of 0.028 - 0.056 g/cm(2) for males and 0.050 - 0.086 g/cm(2) for females, respectively. During their adolescence for males, for every increase of one grade in physical exercise, BMD at varied sites increased by an amount of 0.034 - 0.078 g/cm(2) in males and BMD at lumbar vertebra in males aged 15 - 24 correlated negatively with their total amount of alcohol drinking, and that at lumbar vertebra and of the whole body increased by about 0.035 g/cm(2) in females, which correlated negatively with their total amount of cigarette smoking. Female BMD at lumbar vertebra correlated reversely with their age of menarche and correlated positively with their average time of breast feeding. CONCLUSION: Increase in physical exercise during puberty could increase PBM for the adolescents and keeping adequate body weight and reducing cigarette smoking and alcohol drinking would benefit for the prevention from age-related osteoporosis. PMID- 11864493 TI - [Relationship between use of antibiotics and chronic diarrhoea in infants and young children]. AB - OBJECTIVE: To study the relationship between use of antibiotics and chronic diarrhea in infants and young children. METHODS: A matched case-control study was carried out in Fuzhou City, Fujian Province during April to October 1997, with a total of 32 cases, (aged 1 - 19 months), with chronic diarrhea (with a length > 2 months), and 128 cases of acute diarrhea (as control group I) and 94 cases of acute respiratory tract infection (as control group II), from the Department of Pediatrics, the First Affiliated Hospital of Fujian Medical University during January 1987 to December 1996. RESULTS: Unconditional multiple logistic regression analysis showed that unreasonable use of antibiotics in treatment for acute diarrhea was the major risk factor for chronic diarrhea (OR = 5.61, 95% CI of OR = 1.15 - 27.36 with control group I, and OR = 16.92, 95% CI of OR = 2.67 - 107.32 with control group II). Chi-square test for trend showed that odds ratio of the use of antibiotics in the cases to the controls increased with the number of antibiotics used in treatment. CONCLUSION: Unreasonable use of antibiotics in treatment for acute diarrhea was an important factor contributing to chronic diarrhea in infants and young children. Early pathogenic diagnosis for diarrhea and reasonable use of antibiotics played important roles in prevention form chronicity of diarrheal diseases. PMID- 11864494 TI - [Effects of fasting on expression of insulin-like growth factor-I and IGF binding protein-1 genes in the liver of rats]. AB - OBJECTIVE: To investigate the molecular mechanisms of body development and growth regulated by nutritional status. METHODS: Effects of fasting on the expression of insulin-like growth factor-I (IGF-I) and IGF binding protein-1 (IGFBP-1) genes were analyzed by Northern blotting technique in rats. RESULTS: Fasting for 48 hours, body weight of the rats decreased by 11.8%, level of blood glucose by 50.8% and content of IGF-I mRNA in the liver by 32.7% (P < 0.05). On the contrary, content of IGFBP-1 mRNA in the liver of fasted rats increased gradually, with a 2.5 time increase after 24 h fasting (P < 0.05). There was no significant change in the level of liver albumin mRNA after fasting. CONCLUSION: Fasting could inhibit the expression of IGF-I gene and enhance the expression of IGFBP-1 gene. PMID- 11864495 TI - [A study on factors contributing to intelligence in 857 children]. AB - OBJECTIVE: To study the main environmental factors affecting intelligence of children. METHODS: Intelligence quotients (IQ) were examined in 857 pupils of grades 1 to 6 in a primary school and categorized into four different classes. Data of relevant environmental factors were collected with questionnaires. Uni variate and multivariate logistic regression analyses were performed to identify the factors influencing their IQ. RESULTS: There were 27 factors with statistical significance in uni-variable analysis. And, then they were analyzed with stepwise logistic regression method. Six variables, i.e., parents' education level, breast feeding during infancy, early education at home, nutritional status before three years old, single parent family and average scores at school, enter the regression model finally. CONCLUSION: Intelligence development in children was affected by a lot of factors, and good acquired environmental factors, especially their early nutrition, family background and education, would benefit for their intelligent development and improvement of their intelligent ability in children. PMID- 11864496 TI - [Inhibitory effects of recombinant human neurotrophin-4/5 protein on neurotoxicity caused by arsenic trioxide]. AB - OBJECTIVE: To study preliminarily the inhibitory effects of recombinant human neurotrophin (hNT)-4/5 protein on neurotoxicity caused by arsenic trioxide (As(2)O(3)). METHODS: Nerve cells of the chicken embryonic forebrain and PC12 cells were co-cultured with recombinant hNT-4/5 protein and As(2)O(3) (0 - 16 micromol/L) for 24 and 48 hours, respectively, to observe the survival of nerve cells and the outgrowth of PC12 cells, with clone expression and recombination techniques. RESULTS: There was significant difference in survival rate of nerve cells between the experimental and control groups (P < 0.01) after co-culture for 48 hours, and survival rate and number of neuron outgrowth increased with the concentration of hNT-4/5 protein. CONCLUSION: Recombinant hNT-4/5 protein could inhibit neurotoxicity caused by As(2)O(3), which provided a basis to find an anti toxic factor in the environment by genetic engineering. PMID- 11864497 TI - [Relationship between bone mineral density and blood and urine biochemical indices in women]. AB - OBJECTIVE: To study the relationship between bone mineral density (BMD) and some biochemical indices in their blood and urine of women. METHODS: Fasting urinary calcium (Ca) and creatinine (Cr), ratio of Ca to Cr, urinary hydroxyproline (Hyp), ratio of Hypr to Cr were determined in 163 healthy women, and activity of serum alkaline phosphatase (AKP) were determined in 182 healthy women. BMD at the 1/10 lower end and 1/3 middle-lower site of the radius were measured with single photon absorptiometry for all of them. RESULTS: Serum AKP and ratio of urinary Hyp/Cr increased with age, especially postmenopause, and correlated negatively with BMD. Fasting urinary Ca/Cr ratio also increased with age, especially postmanopause, but not correlated with BMD. CONCLUSION: Serum AKP and fasting urine Hyp/Cr ratio could be used as indicators to reflect bone remodeling and absorption, respectively. PMID- 11864498 TI - [Antagonistic effects of metallothionein on genetic damage caused by gamma-rays and mitomycin C]. AB - OBJECTIVE: To study the antagonistic effects of metallothionein (Zn-MT)on genetic damage caused by gamma-rays and mitomycin C (MMC) in the liver of rabbits. METHODS: Micronucleus test and DNA strand breaks were determined in vitro and in vivo with single-cell gel electrophoresis assay to observe the effects of Zn-MT on the micronucleus formation and cleavage of DNA chain before and after gamma rays radiation and MMC treatment. RESULTS: A dose of 664 microg/kg of Zn-MT could inhibit micronucleus formation in the bone marrow of the mice induced by 5 Gy gamma-rays radiation (P < 0.05). Concentrations of 10 and 50 microg/ml Zn-MT could antagonize the increase in bimicronucleus frequency in g12 cells induced by 1 and 3 Gy gamma-rays radiation and 0.3 microg/ml MMC treatment (P < 0.05 or P < 0.01). A dose of 50 microg/ml Zn-MT could reduce the DNA single strand breaks caused by 1 Gy gamma-rays radiation, but without statistical significance (P > 0.05). CONCLUSION: Zn-MT could antagonize the genetic damage to certain extent in the liver of the rabbits caused by gamma-rays radiation and MMC treatment. PMID- 11864499 TI - [An analysis of nutritional and harmful components of vegetables grown in plastic greenhouses]. AB - OBJECTIVE: To study the changes in nutritional and harmful components of vegetables grown in plastic greenhouses. METHODS: In plastic greenhouses, microclimate and air concentrations of carbon monoxide, carbon dioxide, fluoride and respirable particulate were measured, and chlorophyll, total sugar, crude fiber, nitrite, fluoride, arsenic and some mineral elements in vegetables were determined as compared with those grown in the open-air fields. RESULTS: Greenhouse appeared a lower wind speed and darker illumination. Contents of chlorophyll a an b, total chlorophyll, reduced vitamin C, crude fiber in vegetables grown in greenhouse all were lower than those grown in open-air fields. Contents of potassium, calcium, magnesium, iron, zinc, copper and phosphorous were all lower in the vegetables grown in greenhouse than those grown in open-air fields. The contents of chlorophyll reducing Vitamin C. CONCLUSION: Lower wind speed and inadequate illumination in greenhouse affected photosynthesis and uptake of water in vegetables causing changes in their nutritional components. But, no contamination of burning coal was found in vegetables grown in greenhouse. PMID- 11864500 TI - [Comparison of subclinical infection rates between vaccinated group with type I inactivated vaccine against hemorrhagic fever and controls]. AB - OBJECTIVE: To compare subclinical infection rate in the vaccinated group with type I inactivated vaccine against hemorrhagic fever with renal syndrome (HFRS) with that in the controls and to understand its enhancement. METHODS: A trial field was selected in Jiande County, Zhejiang Province during July 1974 to November 1997. Paired-sera before and after vaccination were collected from 401 vaccinee and 360 controls, respectively. Serum titer of indirect immunofluorescent IgG antibody (IFAT) against HFRS was detected for each of them, and its cut-off value depended on the distribution of serum antibody titer in the second determination in the controls, which could be used to evaluate subclinical infection before vaccination. RESULTS: There was no significant difference in subclinical infection between those with positive and negative IFAT before vaccination, with different cut-off values for identifying subclinical infection. In both vaccinated and control groups with negative IFA before vaccination, subclinical infection rate was 7.62% in the controls, and 2.17% and 1.63% in the vaccinated ones, with cut-off values of 1:160 and 1:320, respectively, significantly different from that in the former. Subclinical infection rate was 11.38% and 6.78% in the vaccinated ones, as cut-off values of 1:20 and 1:40, respectively, without significant difference from the controls. CONCLUSION: No increase in subclinical infection in the vaccinated group with type I inactivated vaccine against HFRS was found. PMID- 11864501 TI - [Relationship between glutathione S-transferase M1, T1 genotype and susceptibility to gastric cancer]. AB - OBJECTIVE: The objective of this study was to examine the association between glutathione s-transferase M1, T1 genotype and the risk of gastric cancer. METHODS: A case-control study was carried out and polymerase chain reaction (PCR) technique was used to identify GST M1, GST T1 genotype in 95 cases of primary gastric cancer and 94 controls. RESULTS: The frequency of GST M1 null genotype was 63.16% and 45.74%, respectively in gastric cancer cases and control group, with statistically significant difference (chi(2) = 5.75, P = 0.0165). GST M1 null genotype correlated with the susceptibility to gastric cancer (OR = 2.03, 95% CI of OR = 1.09 - 3.80). The risk for gastric cancer in those with GST M1 null and GST T1 non-null genotype was significantly higher than that in those with GST M1 non-null and GST T1 null genotype, with an OR of 2.91 and 95% CI of ORs 1.09 - 7.89. The risk for gastric cancer in those smoking with GST M1 null genotype increased significantly (OR = 8.06, 95% CI of OR = 2.83 - 23.67). CONCLUSION: GST M1 null genotype was associated with the risk for gastric cancer. PMID- 11864502 TI - [Evaluation of DNA recombinant chitinase fusion protein antigens in serological diagnosis for filariasis]. AB - OBJECTIVE: To evaluate DNA-recombinant chitinase fusion protein antigen in serodiagnosis for Bancroft's filariasis in jirds rats with microfilaremia and normal jirds rats with ELISA, and its prospect in the market to be spread. METHODS: Oligonucleotide was synthesized with filaria monoclonal antibody (Mf(1)) recognized microfilaria chitinase fusion protein and basic primer recognized by original clonal nucleosides, and amplified with polymerase chain reaction (PCR). PCR products were expressed in Pmal-c vector and purified by affinity chromatography. Experimental animals were inoculated with chitinase fusion protein antigen and infected with larval stage filaria Brugia malayi, respectively. Functional protein molecule of chitinase antigen was analyzed with SDS-PAGE and western blot. Serum level of specific antibody to chitinase antigen was determined by ELISA, and compared with that to microfilaria xt antigen (Mf xt). RESULTS: Specific function protein molecule of the above-mentioned chitinase antigen has been identified as 69,000. Serum antibody against chitinase antigen was 100% positive for jirds and patients with microfilaremia, but, that against Mf-xt only 80% positive. Serum antibody against chitinase antigen was negative for all normal jirds and normal persons, but that against Mf-xt had 5% false positive in normal serum. For those with microfilaremia from non-filariasis prevalent areas, false sero-positivity was 5% and 20% with chitinase antigen and Mf-xt, respectively. CONCLUSION: ELISA with DNA recombinant fusion protein antigen is a sensitive and specific method in serological diagnosis and surveillance for filariasis, with simple, easy and convenient manipulation. It can be used in field serologic surveillance and diagnosis for filariasis during the late phase in control of filariasis and can be spread in good prospects. PMID- 11864503 TI - [Roles of bifidobacterium on prevention of experimental colorectal carcinoma and induction of apoptosis]. AB - OBJECTIVE: To explore the effect of bifidobacterium adolescence on prevention from experimental colorectal carcinoma and its antitumor mechanism. METHODS: An animal model of nude mouse tumor transplanted from colorectal carcinoma was set up. Bifidobacteria adolescence were intraperitoneally injected into nude mice in advance. The growth rate, ultrastructure, density of apoptotic cells, rate of bcl 2 and bax oncoprotein expression and its positive cell density in transplanted tumor of nude mice from colorectal carcinoma were investigated by employing transmission microscopy, in situ end labeling technique and immunohistochemical staining. RESULTS: The growth rate of transplanted tumors from colorectal carcinoma in the group with bifidobacterium prevention was slower than that in the control group. Many tumor cells at varied apoptotic stages could be observed in transplantation tumor tissue in the bifidobacterium prevention group and their density of apoptotic cells, rate of bcl-2 and bax oncoprotein expression and its positive cell density were significantly higher in the group with bifidobacterium prevention, as compared with the control group (P < 0.01). But, expression rate of bcl-2 oncoprotein turned contrary to that of bax oncoprotein. CONCLUSION: Bifidobacteria adolescence could markedly prevent from occurrence and development of colorectal carcinoma in vivo and induce apoptosis of tumor. PMID- 11864505 TI - [Analysis for spatial distribution of Oncomelania snail in mainland China by geographic information system(GIS) database]. AB - OBJECTIVE: To analyze spatial distribution of Oncomelania snail populations in mainland China by geographic information system (GIS) database. METHODS: Genetic variation experiment data and experimental data of susceptibility of 34 snail populations were collected from nine provinces in China, to set up a database. A world digital map was linked with the database in a software ArcView Release 3.0 a to be used to divide zones with ArcView spatial analyze function and GIS overlaying function. AVHRR NDVI satellite images were collected to form a new one for prevalent season, which were overlaid with distribution maps of heterozygous indices of snail populations, polymorphic locus percentage and infection rates of snail population with schistosome for classifying the images. RESULTS: Spatial analysis showed that distribution maps of genetic heterozygous indices of snail, percentage of polymophic loci and infection rate of snail can be divided into two large zones, i.e., east zone and west zone, both with varied pictures. Analysis for overlaying three-stratum distribution map with AVHRR NDVI satellite image showed four distinctly different spatial zones, including west Sichuan zone, west Yunnan zone, middle zone of river, lake and marshland, and southeast coastal zone. CONCLUSION: It is the first time using GIS to analyze data of Oncomelania population genetics and confirmed the population structure of Oncomelania snails presented discrete sub-population model, which supports the theory of sub-species of Oncomelania spp. Exited in mainland China. PMID- 11864504 TI - [A study on immunogenicity and safety of bivalent inactivated vaccine against hemorrhagic fever with renal syndrome]. AB - OBJECTIVE: To study side effects and effectiveness of bivalent inactivated vaccine against hemorrhagic fever with renal syndrome in population of a randomized controlled field trial. METHODS: Serum indirect immunofluorescent antibody in 167 persons and neutralization antibody (types I and II) in 69 persons, who received three-dose vaccine, were determined two weeks after immunization, and side effects in 657 vaccinees were observed within 72 houses after immunization. RESULTS: Serum positive conversion rate of indirect immunofluorescent antibody was 99.04% (166/167) with GMT of 24.51 +/- 2.06. Serum positive conversion rate of neutralization antibody was 100% (69/69), 91.30% (63/69) for types I and 88.41% (61/69) for type II, respectively. GMTs for type I and II neutralization antibody were 18.27 +/- 2.21 and 12.47 +/- 2.16, respectively. Side effects at local site vaccinated of the body and temperature rising could be seen in 1.48% and 0.36% of the vaccinees, respectively. No severe side effect and abnormal reaction was found in vaccinees on the field. CONCLUSION: Immune response to bivalent inactivated vaccine against hemorrhagic fever with renal syndrome was satisfactory with slight side effect. PMID- 11864506 TI - [A study on relationship between hypertension and polymorphism of ACE gene in male Yi people in Liangshan Yi autonomous prefecture, Sichuan]. AB - OBJECTIVE: To explore relationship between polymorphism of ACE gene and hypertension. METHODS: A cross-sectional study was conducted and PCR technique was used to analyze gene polymorphism. RESULTS: Genotypes DD, ID and II of ACE gene accounted for 13.0% and 9.7%, 50.9% and 48.4%, and 36.1% and 41.9%, respectively, in the normotensives and hypertensives. Frequencies of I and D alleles for ACE gene were 61.6% and 66.1%, and 38.4% and 33.9%, respectively, in the normotensives and hypertensives. There was no significant difference in distribution of II, ID and DD genotypes and I/D allele frequencies of ACE gene between the hypertensives and normotensives in male Yi people. CONCLUSION: Association between polymorphism of ACE gene and hypertension was uncertain. PMID- 11864507 TI - [Hemorrheological status in patients with silicosis and its clinical significance]. AB - OBJECTIVE: To study relationship between silicosis and hemorrheology and its clinical significance. METHODS: Varied indices for hemorrheology were measured in 85 male patients with silicosis, 45 male patients with chronic bronchitis and 100 healthy male subjects. RESULTS: Compared with the control group and the chronic bronchitis patient group, average hematocrit (Hct), whole blood apparent viscosity (eta(a)) at 4 s(-1) - 200 s(-1) shear rates, plasma viscosity (eta(p)), erythrocyte electrophoresis time (EET) and platelet electrophoresis time (PET) all significantly increased in patients with silicosis (P < 0.001). Hemorrheological indices in patients with silicosis increased with severity of disease, increased with decrease in their pulmonary function, and were all significantly higher than those in the control group (P < 0.01 approximately equal 0.001). Stepwise regression analysis showed that severity of disease correlated closely to Hct, eta(a) at 4 s(-1) - 200 s(-1), EET and PET, and that pulmonary function status in patients with silicosis and their courses correlated closely to Hct, eta(a) at 4 s(-1) - 200 s(-1), eta(p), EET and PET. CONCLUSIONS: Blood of patients with silicosis was in highly viscose status, which was easy to be complicated with multiple diseases. PMID- 11864508 TI - [Studies on male reproductive toxicity caused by hexavalent chromium]. AB - OBJECTIVE: To study male reproductive toxicity caused by hexavalent chromium (Cr(VI +)). METHODS: Morphology of semen and spermatozoa was observed and chromium level in blood, chromium and zinc level in sperm plasma, leuteinizing hormone (LH) and follicle stimulating hormone (FSH) levels in serum were determined for male workers exposed to (Cr(VI +)) (CrO(3)X = 0.0195 mg/m(3)) with occupational epidemiologic investigation and laboratory analysis. RESULTS: For the exposed workers, their seminal counting was (52.21 +/- 45.51) x 10(9)/L, and zinc level in seminal plasma (4,811.85 +/- 1,401.88) micromol/L, significantly lower than those in controls (88.96 +/- 74.82) x 10(9)/L and (5 718.49 +/- 2 827.90) micromol/L, respectively, P < 0.05. Serum FSH was (7.34 +/- 6.88) IU/L in the exposed groups, significantly higher than that of controls (2.41 +/- 1.69) IU/L, P < 0.01. There were no significant difference in semen volume, semen liquefaction time, prevalence of teratospermia, serum Cr(VI +) and LH level in seminal plasma between exposed group and control group (P > 0.05). Prevalence of teratospermia was 0.24 +/- 0.09, and serum LH and FSH were (7.94 +/- 2.67) IU/L and FSH (9.33 +/- 6.47) IU/L, respectively, in workers exposed to high concentration of Cr(VI +) (CrO(3)X = 0.2351 mg/m(3)) significantly higher than those exposed to lower concentration of Cr(VI +) (CrO(3)X = 0.0172 mg/m(3)), with 0.17 +/- 0.06 of prevalence of teratospermia, (5.05 +/- 3.05) IU/L of LH, and (3.74 +/- 3.04) IU/L of FSH, respectively (P < 0.05). CONCLUSION: The results suggested that damage to convoluted seminiferous tubules epithelium, reduction of spermatozoa formation and increase in prevalence of teratospermia could be caused by exposure to certain concentration of Cr(VI +). PMID- 11864509 TI - [Plasmid DNA breakage caused by reaction of intracellular redox compound with potassium dichromate]. AB - OBJECTIVE: To study plasmid DNA breakage induced by hexavalent chromium (Cr(VI +)) compound in vitro. METHODS: DNA breakage was observed using plasmid relaxation method. RESULTS: pUC 118 plasmid DNA single strand breakage can not be caused by either ascorbic acid (1 mmol/L), or glutathione (5 mmol/L), or hydrogen peroxide (H(2)O(2), 17 mmol/L) or potassium dichromate (K(2)Cr(2)O(7), 0.5 mmol/L) alone. But, if DNA was incubated warmly with K(2)Cr(2)O(7) and intracellular redox compound, pUC118 plasmid DNA single strand breakage could be induced by K(2)Cr(2)O(7), ascorbic acid, or glutathione, or products of peroxidation reaction. K(2)Cr(2)O(7) (0.5 mmol/L) and H(2)O(2) (17 mmol/L) also could cause some double-strand breakage in plasmid. CONCLUSION: It suggests that ascorbic acid, glutathione and H(2)O(2) can play an important role in carcinogenicity of K(2)Cr(2)O(7). PMID- 11864510 TI - [Effects of lead exposure on their brain-stem auditory evoked potential in children]. AB - OBJECTIVE: To determine if low lead exposure can influence auditory development in children. METHODS: Children were divided into two groups, i.e., low (< 0.483 micromol/L) and high (>or= 0.483 micromol/L) blood lead levels with atomic spectrophotometry (AAS). Latencies and latent intervals between peak waves of their brain-stem auditory evoked potentials (BAEP) in the two groups were measured and compared. RESULTS: There was positive correlation between blood lead levels and BAEP in wave III at right ear, wave V at both right and left ears and latent intervals between peak waves I to V, with correlation coefficients of 0.2991, 0.2748, 0.3248, 0.2064 and 0.2412, respectively. There was significant difference in latencies of wave III at right ear, wave V at both right and left ears and latent intervals between peak waves I to V at both right and left ears and those between waves of III to V at right ear in the low blood lead group. Latencies and latent intervals between peak waves were longer in the high blood level group (with a mean of 0.78 micromol/L and a range of 0.58 to 1.42 micromol/L) than those in the low one (with a mean of 0.30 micromol/L and a range of 0.17 to 0.45 micromol/L). CONCLUSION: Elevated blood lead level could cause prolongation of latencies and latent intervals between peak waves of BAEP and slowdown of the nerve conduction velocity, and so auditory development was influenced. PMID- 11864511 TI - [Studies on DNA single strand breaks induced by sodium nitroprusside-nitric oxide donor]. AB - OBJECTIVE: To study the effect of sodium nitroprusside (SNP), a nitric oxide (NO) donor, on DNA single strand breaks (SSBs). METHODS: A modified method was used to isolate and prepare the single cell suspension from the organs of mice. Alkaline single-cell gel electrophoresis (SCGE) was performed to examine DNA damage of the cells treated by SNP in vivo and in vitro. RESULTS: Treatment with 0.5 - 2.0 micromol/ml of SNP with S(9) for 1 h induced a concentration-dependent increase in DNA SSBs in g12 cells. Significant increase in DNA migration and comet frequency in the spleen, thymus and peritoneal macrophage were induced after intraperitoneal injection of SNP at a dose of 0.67 - 6.0 mg/kg. No obvious increase in DNA single strand breaks was observed in the liver, kidney and lung of the mice with same treatment. CONCLUSION: DNA SSBs could be induced by NO in some cells in vivo and in vitro. There was difference in sensitivity of various organs in the mice to NO. Cells of spleen and thymus and macrophage may be the important target cells of NO. PMID- 11864512 TI - [A study on formation of N-(nitrosomethyl) urea in experimental pig stomach gavaged with fish sauce]. AB - OBJECTIVE: To study formation of N-(nitrosomethyl) urea (NMU), which is categorized as N-nitrosamides -- a sort of strong chemical carcinogen, by perfusion with fish sauce via pig stomach fistula, which is used daily as cooking flavor by local residents in Changle County, Fujian Province, a highly-prevalent area for gastric cancer. METHODS: Fistulization was performed in fasting experimental pigs' stomach, and their gastric juice was suctioned 30 minutes after perfusion with fish sauce and sodium nitrite via fistula. Gastric juice specimen was purified and concentrated and determined for NMU with high performance liquid chromatography-photolysis pyrolysis-thermal energy analyzer. RESULTS: NMU was detected in the pig stomach at pH 1 - 2, where NMU formed. Formation of NMU depended on the amount of nitrite added in a dose-dependent relationship. Level of NMU in gastric juice reached 25.4 and 7.97 micromol/L, respectively, when 3.48 and 0.87 mmol of sodium nitrite were fed into the stomach. No NMU could be detected, as 0.22 mmol of sodium nitrite fed. CONCLUSION: Under the condition of pH 1 - 2 and presence of sodium nitrite, NMU was synthesized in the stomachs of experimental pigs fed with fish sauce via gastric fistula. It is postulated that N-nitrosamides, such as NMU could be formed endogenously in the stomachs of local residents who consume fish sauce often and have a higher exposure to nitrite in their stomachs. PMID- 11864513 TI - [Induction of quinone reductase and glutathion-s-transferase by dimethyl fumarate in rats]. AB - OBJECTIVE: To assess induction of quinone reductase (QR) and glutathion-s transferases (GSTs) by dimethyl fumarate (DMF) in major viscera of rats. METHODS: Fodder fed to male Wistar rats was supplemented with 2.0% DMF, and blood specimens were collected form the rats 6 h, 24 h, 72 h, 1 week and 2 weeks after feeding, respectively, and then animals were killed. QR and GSTs activities were determined in rats' gland stomach, liver, lung, kidney and heart, and also QR, LDH and GPT activities in their sera were measured with enzyme dynamic methods. RESULTS: Administration of DMF for 24 h, 72 h, 1 week and 2 weeks in rats can significantly increase activities of QR and GSTs in their gland stomach, liver, lung and kidney, as compared with those in controls (P < 0.0001, or P < 0.01), and their serum QR also significantly increased (P < 0.0001, or P < 0.01). But, DMF had no effect on activities of serum GPT and LDH (P > 0.05). CONCLUSION: DMF is a potent induction agent for QR and GSTs in the gland stomach, liver, lung and kidney of rats, which suggests it will hopefully be a protective and antidotic agent against human antioxidant injury. PMID- 11864514 TI - [Effects of different levels of calcium intake on bone of pregnant women]. AB - OBJECTIVE: To research some effects of different levels of calcium intake in the mother's bone mineral density. METHODS: On the basis of insignificantly different among energy and other nutrition intake excepting calcium, 35 volunteers in pregnant 18 weeks healthy women were randomly divided into I. II and III group [calcium intake were (550 +/- 150) mg/d, (900 +/- 150) mg/d and (1,500 +/- 150) mg/d]. The tracked study began from pregnant 20 weeks to postpartum 45 days. All over and different body of BMD (Bone Mineral Density) were assessed by dual energy X-rays absorptiometry. RESULTS: I group of many location BMD were below that of other groups (P < 0. 01). As compared with the same age women's lumbar spine BMD, I group was only maintained within the rang (85.14 +/- 6.61)%, and osteoporosis in some places of bone were found in some individual. II group was (90.74 +/- 6.53)%. III group was (100.44 +/- 5.19)%. Allover different body's BMD were significantly correlated with the average BMD (r > 0.8, P < 0.1), the most significant correlation was third lumbar spine. Calcium intake of women was positively correlated with different body's BMD (r > 0.5, P < 0.01). CONCLUSION: III group, calcium intake was (30.68 +/- 3.56) mmol/d, pass through period of pregnancy, parturition and short-term breast-feed mother body remain the same BMD, and prevent osteoporosis. Reporting pregnant women calcium nutrition pass through BMD, lumbar spine is good representation. PMID- 11864515 TI - [Neonatal screening and collaboration among scientific institutions]. PMID- 11864516 TI - [Predictive factors of efficacy in antimicrobial prophylaxis for acute recurrent otitis media]. AB - OBJECTIVE: A prospective study was designed to assess the most relevant prognostic factors influencing outcome in children treated with antibiotics for recurrent acute otitis media (AOM). MATERIALS AND METHODS: A prospective study was performed in 69 patients with recurrent AOM who received prophylactic treatment in the Pediatric Otorhinolaryngology Section of our hospital between January 1998 and May 1999. In all patients, 17 variables were registered: age, sex, family history of recurrent AOM, history of prematurity, history of allergies, use of day care centers, household smokers, maternal smoking, coincidence with dental eruption, symptoms of upper respiratory obstruction (snoring, breathing through the mouth, obstructive tonsils), episodes of tonsillitis per year, age at first AOM episode, season in which prophylaxis was started, and characteristics of neonatal feeding (breast or bottle feeding and position). ResultsThe mean age was 35.3 months (range: 9-120 months). There were 38 boys (55.1 %) and 31 girls (44.9 %). The factors related to a more favorable response to treatment were lower age at the first AOM episode (p 0.004) and a positive family history of AOM (p 0.047). CONCLUSIONS: The children that benefitted most from the prophylactic schedules for recurrent AOM were those that had the first episode at an earlier age and who had a positive family history of the disease. PMID- 11864517 TI - [Septic arthritis due to Streptococcus pneumoniae]. AB - INTRODUCTION: There are few reports of Streptococcus pneumoniae as a cause of septic arthritis. The aim of this study was to describe our experience with five documented cases of pneumococcal arthritis over 14 years. METHODS: The medical histories of infectious septic arthritis in our center between January 1986 and December 2000 were retrospectively reviewed. Cases of pneumococcal etiology were analyzed while those occurring in the neonatal and immediate postoperative periods were excluded. RESULTS: Among the 39 documented cases of septic arthritis, 5 (12.8 %) were caused by Streptococcus pneumoniae. Four of the patients were younger than 15 months old. Fever without localizing signs as an antecedent was present in four patients; of these, one patient had concurrent otitis media. The most common localization was the hip (four patients) and the knee (one patient). In three patients simple radiology showed no abnormalities while in four ultrasonography showed abnormalities. Etiologic diagnosis was established by isolation of S. pneumoniae from blood culture and synovial fluid (one patient), from blood culture (one patient), from synovial fluid (two patients), and by detection of pneumococcal antigen in urine (one patient). Of the germs isolated, two showed reduced susceptibility to penicillin (CMI 0.125-1 mg/l) and all were susceptible to cefotaxime. Open drainage was performed in all patients with hip involvement. None of the patients presented sequelae. CONCLUSIONS: In our experience, S. pneumoniae should be taken into account as a cause of septic arthritis that mainly affects children younger than 15 months. The hip was the most frequently involved joint. Joint fluid culture was the method providing the best diagnostic yield. A high proportion of strains are not susceptible to penicillin. Early diagnosis and management lead to a good prognosis. PMID- 11864518 TI - [Renal failure as a factor leading to epileptic seizures]. AB - OBJECTIVE: To analyze the incidence of convulsive seizures in renal failure by studying the semiological characteristics of the convulsive episodes, their subsequent evolution and the presence of possible sequelae. The relationship between the different potential causes of the seizures and their evolution and treatment was also analyzed. MATERIAL AND METHODS: We retrospectively reviewed the clinical histories of 108 patients admitted to the hospital over a 20-year period with a diagnosis of renal failure: 55 were undergoing predialysis, 42 renal transplantation, 7 peritoneal dialysis and 3 hemodialysis. One patient was excluded for not fulfilling the selection criteria. Computer study of the 18 quantitative and qualitative variables was carried out with the SPSS 9.0.1 program. RESULTS: Of the 107 patients, 16 (14.95 %) had suffered some type of convulsive seizure. None was in the predialysis group. As triggering factors, hydroelectrolyte imbalance was found in eight patients and hypertension was found in four. In the remaining patients the causes were not well-defined. Only three patients with epilepsy prior to renal failure presented sequelae. CONCLUSIONS: Epileptic seizures in renal failure in childhood can be considered as occasional seizures that do not usually become chronic or produce sequelae. Information on the management of seizures in renal failure should be disseminated among professionals treating systemic diseases. PMID- 11864519 TI - [Maltreatment of children with disabilities: Characteristics and risk factors]. AB - OBJECTIVES: To determine the prevalence of maltreated children with disabilities, the demographic characteristics, the frequency of the type of maltreatment, the perpetrator of the maltreatment, the family, social and environmental risk factors and to contribute information to knowledge of the etiopathogenesis of child maltreatment. PATIENTS AND METHODS: We performed a retrospective cross sectional study over a 10-year period (1991-2001). The health and socio-familial reports of 1,115 maltreated children entering foster care in Saragossa (Spain) were reviewed and those presenting physical, psychological and/or sensorial disability were included. RESULTS: Sixty-two children with disabilities were admitted during the study period (a prevalence of 5.56 % below that expected). Mean age was 7.47 years (5.60 SDU) (range: 0-17 years). The most frequent age group was 0-5 years (43.5 %). The male-to-female ratio was 1.36:1. The most prevalent disability was psychological (35.5 %), followed by mixed (27.4 %), physical (25.8 %), and sensorial (11.2 %). The most frequent type of maltreatment was physical neglect (82.2 %). A total of 16.2 % of the children presented several types of maltreatment. In 83.8 % of the patients the perpetrator of the maltreatment was the mother. At least one family, social and/or environmental risk factor was present in 93.5 % of the children and more than one risk factor was present in 64.5 %. CONCLUSIONS: We believe that in most of the cases of maltreatment of disabled children from our environment, the etiopathogenesis was unrelated to the presence of an isolated individual risk factor (disability). Instead, maltreatment was related to the existence of familial, social and environmental risk factors. The greater the number of risk factors associated with a disabled child, the greater will be the possibility of maltreatment. PMID- 11864520 TI - [Intraepithelial lymphocytes in celiac disease]. AB - Coeliac disease (CD) is a permanent intolerance to gluten that provokes alterations in the mucosa of the small intestine. The disease can usually be controlled by excluding gluten from the diet. CD is immunologically-mediated, with a strong linkage to certain HLA alleles and a permanently altered intraepithelial lymphocytes (IEL) pattern. The development of a flow cytometric technique for the evaluation of IEL subsets has increased our understanding of these alterations and has prepared the ground for its clinical application. Our experience shows that this procedure has excellent sensitivity and specificity in the diagnosis of CD and that it is particularly useful in the evaluation of atypical presentations of the disease. The present article reviews our experience in the diagnosis of CD and discusses some of the hypotheses that have been put forward on the possible role of IEL in its pathogenesis. PMID- 11864521 TI - [Role of signs of fetal hypokinesia in the diagnosis of spinal muscular atrophy of neonatal onset]. AB - INTRODUCTION: Spinal muscular atrophy (SMA) is characterized by early degeneration of anterior horn cells. The most frequent and severe type of neonatal onset is Werdnig-Hoffmann disease. The neurologic and genetic characteristics of SMA are well-known. The aim of this study was to analyze the dysmorphologic features of this disease. PATIENTS AND METHODS: We present an analysis of 10 cases of SMA identified among 27,864 infants with congenital defects registered by the Spanish Collaborative Study of Congenital Malformations (ECEMC) between April 1976 and December 1998. We also report a clinical case of neonatal SMA presenting the classical signs of fetal hypokinesia deformation sequence. RESULTS: The minimum estimation of the prevalence of SMA with congenital defects in our population is 0.32 per 100,000 live births. We found a male-to-female ratio of 3.5. The most frequently associated congenital defects in our population of neonatal SMA were located in the extremities (mainly arthrogryposis), face and thorax and could be explained by intrinsic fetal hypomobility secondary to the neuromuscular disorder. The characteristics of fetal hypokinesia deformation sequence are discussed in the case report presented herein: dystocic delivery, short umbilical cord, polyhydramnios, intrauterine growth retardation, craniofacial malformations, skeletal abnormalities with multiple articular contractures, pulmonary hypoplasia, etc. CONCLUSIONS: It is important to recognize the congenital defects associated with neuromuscular disorders, because dysmorphologic features are sometimes more marked than neurologic features in the neonatal period and because of the wide spectrum of congenital defects in neonatal SMA that result in a fetal hypokinesia deformation sequence. PMID- 11864522 TI - [Cocaine toxicity in the newborn. Detection and prevalence]. AB - The neonatal effects of fetal cocaine exposure are not completely known. Serious adverse effects have been described in several series of women consuming pure cocaine, but in recent years articles questioning some of the damaging effects of cocaine on the fetus and newborn have been published. The use of cocaine is currently increasing both in Spain and in many other countries and this will in turn lead to an increase in the incidence of fetal and neonatal adverse effects. We review the possible effects of cocaine exposure in utero, and place particular emphasis on neurobehavioral abnormalities, which are present even when exposure is limited to the first trimester of pregnancy. We also analyze various biomarkers for cocaine detection in newborns and their mothers and, finally, we review pharmacogenetic and dose-response relation susceptibility factors. PMID- 11864523 TI - [Metachronous bilateral Wilms' tumor]. AB - Wilms' tumor occurs in 5-10 % of all cases of nephroblastoma. The metachronous form represents 2-3 % of cases. Most (96.2 %) metachronous tumors appear within the first 5 years of the primary tumor. Associated malformations are more common in bilateral cases. Metachronous tumors are a therapeutic challenge. We describe the case of an 11-year-old girl with left hemihypertrophy. The diagnosis was metachronous relapse of Wilms' tumor 7 years after the first diagnosis. The patient received five courses of preoperative chemotherapy and tumorectomy was performed. Because of post-surgical complications, nephrectomy was performed on her only kidney. Since she is anephric, the patient is in chronic renal failure and is dependent on dialysis. Treatment with carboplatin and etoposide was continued after surgery and the patient is currently in complete remission. The appearance of a metachronous Wilms' tumor 5 years after that of the primary tumor is rare. When a contralateral tumour develops, chemotherapy must be given until the size of the tumor is reduced in order to preserve renal function and avoid dialysis. In patients with chronic renal failure caused by bilateral nephrectomy, ongoing treatment with dialysis support can be achieved through the choice of effective drugs and knowledge of their pharmacokinetics and pharmacodynamics. PMID- 11864524 TI - [Group B streptococcus cellulitis-adenitis syndrome in neonates. Is it a marker of bacteremia?]. AB - We report the case of an infant in whom facial cellulitis was the only initial sign of a group B streptococcal bacteriemia. Adenitis-cellulitis syndrome is a rare clinical manifestation of group B streptococcal infection in infants. Local infection together with the onset of fever can be the only signs of bacteriemia in newborns and infants. PMID- 11864525 TI - [Hypohidrotic ectodermal dysplasia: A cause of fever of unknown origin]. AB - The term ectodermal dysplasias includes many disorders that share some clinical features such as involvement of one or several ectodermal structures and congenital origin. Currently, 154 different types divided into 11 clinical subgroups (Freire Maia classification 1994) have been described. The most frequent entity is hypo- or anhidrotic ectodermal dysplasia (Christ-Siemens Touraine syndrome). This is a rare hereditary disease whose main characteristic is the absence, or more often the reduction, of sweat glands, leading to an increase in body temperature together with anomalies of the epidermis and its appendages (hair and nails). We present a case of hypohidrotic ectodermal dysplasia in a premature 18-month-old boy who was referred to our department because of markedly dry skin since birth and recurrent eczematous and lichenification lesions that had been successfully treated with topical corticosteroids. Physical examination revealed mild alopecia with sparse and fine blonde hair and the absence of dental alveoli. The boy's mother had noticed slight sweating and episodes of fever without clinical symptoms, which were more frequent in summer. Hypohidrotic ectodermal dysplasia should be included in the differential diagnosis of fever of unknown origin. PMID- 11864526 TI - [Congenital colonic stenosis]. AB - The colon is the least common site of congenital intestinal stenosis and atresia and accounts for 5-15 % of all of these abnormalities. We present the case of a 2 month-old boy who presented vomiting, constipation, weakness, and abdominal distension. Contrast enema and colonoscopy revealed an abrupt change in caliber in the sigmoid colon and, after ruling out other disorders, a diagnosis of colonic stenosis was made. Laparoscopy under total anesthesia was performed and a stenotic area in the sigmoid colon and proximal dilatation were identified. A small incision was made and the stenotic segment was removed. Video-assisted resection and end-to-end anastomosis were performed and the child made and uneventful postoperative recovery. Contrast enema after surgery showed good anastomosis and normal bowel diameter. At the age of 18 months the patient is asymptomatic and his stools are normal. With laparoscopy, visualization of the abdominal cavity was magnified and detailed, the affected intestinal segment was easily identified and other diseases were ruled out. Recovery was rapid and the functional and esthetic results were excellent. PMID- 11864527 TI - [Extralobar pulmonary sequestration with supernumerary esophageal bronchus]. PMID- 11864528 TI - [Endoluminal expandable stents for severe tracheobronchomalacia]. PMID- 11864529 TI - [Mycoplasma pneumoniae pneumonia]. PMID- 11864530 TI - [Prenatal cystic neuroblastoma]. PMID- 11864531 TI - [Budd-Chiari syndrome]. PMID- 11864532 TI - [Ramsay-Hunt syndrome]. PMID- 11864533 TI - [Spastic monoparesis]. PMID- 11864535 TI - [Percutaneous intrahepatic portosystemic shunting as a treatment for refractory hepatic hydrothorax]. AB - Hepatic hydrothorax is an infrequent complication of portal hypertension in liver cirrhosis. Treatment with saline restriction and diuretics is usually effective but when this fails, the therapeutic approach is difficult and multiple complications occur. Transjugular percutaneous intrahepatic portosystemic shunt (IPS) is associated with a marked decrease in portal pressure and consequently this technique has been used in the treatment of refractory ascites. The aim of this study was to analyze the efficacy, safety and outcome of refractory hepatic hydrothorax treated by IPS. The procedure was performed in 5 patients who were all grade B or C in the Child-Pugh classification. Three patients showed complete response to the treatment, of whom 1 underwent transplantation 20 days later. The fourth patient showed partial response with a reduction in the need to perform thoracocentesis and the fifth patient showed no response to IPS and died after 17 days of follow-up. Albumin levels and Child classification remained unchanged. Two patients presented recurrence with reappearance of hydrothorax due to shunt dysfunction and 2 patients presented hepatic encephalopathy that responded to medical treatment. Refractory hepatic hydrothorax can be controlled by IPS in a large number of patients but its efficacy is restricted by shunt dysfunction, the risk of encephalopathy and by its limited effect on survival. PMID- 11864536 TI - [Hemodynamic modifications in splenic circulation studied by echo-Doppler during pregnancy]. AB - INTRODUCTION: Pregnancy produces multiple changes in the mother's body, most of which have been studied. However, changes in hepatosplenic circulation are not well-known. The routine use of ultrasonography and of echo-Doppler has created new possibilities for the knowledge of splenic circulation during pregnancy. MATERIAL AND METHOD: We studied 30 healthy pregnant women who had given their informed consent. To evaluate the morphological and hemodynamic changes that might occur in the splenic vessels during pregnancy and immediate puerperium, laboratory investigations, obstetric and hepatic ultrasonography, and hepatosplenic echo-Doppler were performed between weeks 8-12, 20-24, and 32-36, as well as in the immediate puerperium. RESULTS: The caliber of the vena porta and its tributaries, as well as that of its intrahepatic branches, increased while the caliber of the suprahepatic vessels slightly decreased during pregnancy. The hemodynamic changes detected by Doppler ultrasonography were: progressive flattening of the pulsed Doppler trace of the suprahepatic vessels during the course of pregnancy; a progressive reduction in mean portal velocity, which was more marked in the third trimester, and a decrease in the markers of resistance in the hepatic artery and superior mesenteric artery. CONCLUSIONS: Hemodynamic changes in hepatosplenic circulation are produced during pregnancy that can be safely and effectively evaluated in real time by ultrasonography and echo-Doppler. Knowledge of these changes is required to evaluate these vessels in pathological conditions. PMID- 11864537 TI - [Coadjuvant combined treatment of hepatocellular carcinoma prior to liver transplantation]. AB - The aim of this study was to report the antitumor effect of combination therapy of hepatocellular carcinoma in patients on the waiting list for liver transplantation. We studied 3 cirrhotic patients with hepatocellular carcinoma > 3 cm and < 8 cm who underwent transarterial embolization and percutaneous ethanol injection as combination coadjuvant therapy while on the waiting list for liver transplantation. Transarterial embolization failed to produce total necrosis of the tumor. In all 3 patients this was subsequently achieved with percutaneous ethanol injection. All the patients currently remain on the waiting list and show no signs of local tumor recurrence 9, 10 and 13 months after the procedures. In conclusion, combination adjuvant therapy with transarterial embolization and percutaneous ethanol injection can increase tumor necrosis and can be useful prior to liver transplantation. Further studies with a greater number of patients are required to confirm the value of this combination therapy. PMID- 11864538 TI - [Gastrointestinal alterations in a female patient with common variable immunodeficiency]. AB - We present the case of a 33-year-old woman who complained of intermittent diarrhea over the previous 8 years and who was diagnosed with common variable immunodeficiency. The woman presented antral atrophic gastritis, partial atrophy of the duodenal villi, nodular lymphoid hyperplasia of the small intestine and lymphocytic colitis. We also review the literature. PMID- 11864539 TI - [Celiac disease and Bud-Chiari syndrome: an uncommon association]. AB - Celiac disease can present great clinical heterogeneity. Its association with a series of intestinal and non-intestinal diseases, whether immunologically mediated or otherwise, presents a higher than normal frequency. We present a patient with celiac disease and Budd-Chiari syndrome of unknown cause. This association has previously been described only in isolated cases in northern Africa. The appearance of this case in Spain reveals that the coexistence of both processes in a single patient is unlikely to be due to environmental or geographical factors. PMID- 11864540 TI - [Recommendations for the use of infliximab (Remicade) in Crohn's disease. GETECCU 2001]. PMID- 11864542 TI - [Drug related problems as a cause of hospital admission]. AB - BACKGROUND: The goals of the present study were: a) to identify and characterize drug-related problems (adverse drug reactions, non-compliance and intentional overdose) leading to admissions to our hospital (Hospital Clinic) through the Emergency Department, b) to know the most frequently involved medications, c) to identify risk factors for hospital admissions due to drug-related problems, and d) to determine the avoidability of these admissions. PATIENTS AND METHOD: Prospective study carried out from August 1999 to November 1999 and from January 2000 to May 2000. The study included 1,800 admissions corresponding to 1,663 patients. RESULTS: Including definitive, probable and possible cases, 215 hospital admissions (11.9%) were drug-related ones. Of these cases, 108 (50.2%) corresponded to adverse drug reactions, 100 (46.5%) to dose-related therapeutic failures (mainly non-compliance) and 7 (3.3%) corresponded to intentional overdose. When 'possible' cases were excluded in order to achieve a better defined causative relationship, drug-related admissions dropped to 139 (7.7%). According to Schumock and Thornton criteria, 68.4% hospital admissions due to drug-related problems were considered 'avoidable'. Most avoidable admissions were due to non-compliance, absence of preventive therapy and inappropriate monitoring. CONCLUSIONS: Hospital admissions due to drug-related problems are frequent and often preventable. PMID- 11864541 TI - [Diagnostic localization of insulinoma and prognostic value of postoperative glycemia monitoring]. AB - BACKGROUND: Our purpose was to establish the efficacy of different diagnostic tests for the diagnosis and localization of insulinoma as well as the prognostic value of postoperative glycemia measurements. PATIENTS AND METHOD: Retrospective analysis of all insulinoma patients diagnosed at our hospital over the last fifteen years. Every patient underwent a supervised prolonged fasting and localization imaging tests were performed in positive cases. In the immediate preoperative period, during surgical intervention and immediately after it, an intravenous dextrose solution (10% concentration, 2-3 g/Kg/d infusion rate) was used to avoid hypoglycemia. Glycemia was measured every 6-8 h from the immediate postoperative period until hospital discharge. RESULTS: Seventeen insulinoma cases were identified (incidence 2.5 cases/million/year), 59% women and 41% men. The sensitivity (and positive predictive value) of diagnostic tests was: conventional-contrast CT scan 30% (75%), spiral CT scan 71.42% (100%), arteriography 28.57% (100%) and intraoperative ultrasonography 100% (100%). MRI identified insulinoma in two out of five patients and In-pentetreotide scintigraphy only in one out of five. Patients subsequently developing diabetes mellitus or glucose intolerance had higher levels of postoperative glycemia, which also lasted for more days, and longer need for insulin therapy. However, patients with a recurrence of the tumor did not show rebound hyperglycemia at all. CONCLUSION: Postoperative glycemia could be a useful prognostic factor in patients who develop diabetes mellitus or glucose intolerance after resection of insulinoma and to predict insulinoma recurrence. PMID- 11864543 TI - [Cost-effectiveness of hypertension treatment in Catalonia (Spain)]. AB - BACKGROUND: We assessed the cost-effectiveness of pharmacological treatments for hypertension to prevent coronary heart disease and stroke in Catalonia (Spain). METHODS: Cost-effectiveness was measured as the cost in Spanish Ptas per life year gained (LYG) in 1998 in individuals aged 40 to 69 years with moderate/severe hypertension (>= 105 mmHg) and mild hypertension (95-104 mmHg). We evaluated hydrochlorothiazide (diuretic), propranolol (-blocker), nifedipine (calcium antagonist), captopril (angiotensin-converting-enzyme inhibitor) and prazosin (* adrenergic blocker). RESULTS: Cost-effectiveness ranged from 706,100 to 446,780 ptas. per LYG in men and from 635,100 to 810,270 ptas. per LYG in women with moderate/severe hypertension and from 108,770 to 682,460 ptas. per LYG in men and from 101,000 to 12,699,000 ptas. per LYG in women with mild hypertension. Incremental cost-effectiveness analysis showed that hydrochlorothiazide and propranolol were the most cost-effective treatments in individuals with moderate/severe hypertension while hydrochlorothiazide and nifedipine were most cost-effective in those with mild hypertension. CONCLUSION: In this study, greatest-to-lowest cost-effectiveness of assessed treatments was as follows: hydrochlorohiazide, propranolol, nifedipine, prazosin and captopril in moderate/severe hypertension and hydrochlorothiazide, nifedipine, propranolol, prazosin and captopril in mild hypertension. PMID- 11864544 TI - [Iodine deficiency in pregnant and fertile women in an area of normal iodine intake]. AB - BACKGROUND: Our purpose was to evaluate the efficacy of a iodine prophylaxis campaign in pregnant women. PATIENTS AND METHOD: Eighty-one pregnant and 29 fertile women were enrolled. The urine iodine concentration was determined. RESULTS: The median urine concentration of iodine was similar in both groups [pregnant women: 113.3 g/l; non-pregnant women: 99.1 g/l]. 67.9% pregnant women and 51.7% non-pregnant women had urine concentrations of iodine lower than the normal range. CONCLUSIONS: Our results suggest that the ongoing iodine prophylaxis campaign in our community is not effective in the adult population. PMID- 11864545 TI - [Selective inhibitors of cyclooxigenase-2 (Coxibs) and cardiovascular morbidity]. PMID- 11864546 TI - [New strategies and new drugs in the salvage therapy for HIV infection]. PMID- 11864548 TI - [Gastrointestinal and cardiovascular side effects associated with COX-2 selective inhibitors]. PMID- 11864547 TI - [GESIDA/PETHEMA guidelines for the diagnosis and treatment of lymphomas in HIV infected patients]. PMID- 11864549 TI - [Safety of celecoxib and rofecoxib: a critical overview]. PMID- 11864550 TI - [Rofecoxib-induced upper gastrointestinal bleeding]. PMID- 11864553 TI - [Spinal dural arteriovenous fistula: surgical or endovascular treatment?]. PMID- 11864552 TI - [Liver abscess: a rare manifestation of Gemella morbillorum infection]. PMID- 11864554 TI - [Spinal cord dural arteriovenous fistulae: analysis of a series with surgical curative treatment]. AB - BACKGROUND: Spinal dural arteriovenous fistulae (SDAVF) are elusive to an early diagnosis. Otherwise, there is no agreement regarding the best treatment. AIM: To review our series of spinal arteriovenous malformations to illustrate the treatment and outcome of FAVDE. PATIENTS AND METHODS: Descriptive series of cases diagnosed and treated at our hospital within the last ten years. Ten suspicious MRI, followed by angiograms, got the next diagnosis: 3 intraspinal malformations, 2 cavernoms and 5 FAVDE, reported here. RESULTS: Four out of 5 FAVDE were men. Mean age was 61year (53-77), and mean interval to diagnosis 10.8 months (1-24). The clinical course consisted of progressive paraparesis, wich was acute in one case, with wheelchair confinement. Sensory involvement and sphinter dysfunction were also always present; severe pain affected 2 cases. MRI revealed the FAVDE at low thoracic or lumbar levels, with tortuous flow voids over the surface of the affected area. Angiograms disclosed the single feeding vessel between D8-L3. Laminectomy and interruption of the draining vein was the chosen treatment. Postoperative complications were not found. Improvement followed until full recovery, that occurred in 4 cases. One patient, previously embolized, was the only one with just modest recovery, although the angiogram proved that deferred surgery ran well. Mean follow-up was 3.8 years (1.5-6); neither clinical deterioration nor suspicious MR finding were found. Postoperative angiogram, performed in two cases, confirmed the fistulae as occluded. CONCLUSIONS: FAVDE mainly involves old males with progressive paraparesis. Suitable MR findings and characteristic angiograms allow the diagnosis. Novel patients are subsidiary to selective and simple surgery, that is usually curative without recurrencies. PMID- 11864555 TI - [Correlation between reaction time and bradykinesia in Huntington's disease]. AB - BACKGROUND: Bradykinesia is the clinical feature of Huntington's disease most closely related to functional discapacity and stage, probably as a consequence of spreading of neuronal injury. The aim of the present article is to verify whether a choice reaction time could be considered an estimate measurement of clinical bradykinesia, and its possible relationships with other evolutive parameters such as functional discapacity, clinical stage and prefrontal executive dysfunction. METHODS: Fifteen patients were studied (9 in advanced stage and 6 initials), equal number of controls and 3 asymptomatic gene carriers. We used clinical bradykinesia and functional capacity scales, an extensive prefrontal battery and a computerized paradigm of reaction time. RESULTS: Clinical bradykinesia and reaction time are closely related. The associations between reaction time with those parameters indicatives of prefrontal dysfunction, functional discapacity and clinical stage are closer and more significatives than those that could be established with clinical bradykinesia. CONCLUSIONS: Reaction time is an objective measurement of global motor slowness that allow us assigning each subject to a specific stage, and avoid possible errors derived from interobserver bias in clinical scales. PMID- 11864556 TI - [Antineural antibodies and paraneoplastic neurologic syndromes]. AB - The discovery that many paraneoplastic syndromes of the nervous system associate with the presence of specific antineuronal antibodies in the serum and cerebrospinal fluid of patients has facilitated the recognition of these disorders and attracted the interest of clinical and laboratory investigators. In this review, the authors discuss the most common immunological findings associated with paraneoplastic syndromes of the nervous system, and provide a practical clinical review of those associated with antineuronal antibodies. PMID- 11864557 TI - [Generalized muscular dystonia: An historial overview]. PMID- 11864558 TI - [Profile of the antimigraine drug almotriptan]. AB - Triptans are the most relevant advance in the symptomatic treatment of migraine in the last decade. In this paper the basic properties as well as the efficacy and tolerability data of the last launched triptan, almotriptan, are reviewed. Almotriptan exhibits the highest bioavailability among triptans, this absorption being unaffected by sex, the presence of food in the stomach or by a migraine attack. At the recommended dose of 12.5 mg, almotriptan shows a rapid onset of effect, at 30 minutes, with efficacy parameters comparable to those of placebo. The potential advantages of almotriptan seem to be three: recurrence rate in the low range, lack of interactions with other drugs and a placebo-like tolerability profile, with a very low rate of chest symptoms. This balanced profile between efficacy and tolerability makes almotriptan 12.5 mg a good option for the symptomatic treatment of migraine. PMID- 11864559 TI - [Giant vertebral dural ectasia and neurofibromatosis type 1]. PMID- 11864560 TI - [Peripheral multineuritis pointing at systemic sarcoidosis]. AB - We report a patient with a peripheral neuropathy as the first symptom of sarcoidosis. The systemic illness was proved by the presence of typic granulomes in the bone marrow. The fact that sarcoidosis is the cause for the neuropathy is supported by the temporary relation and by the good response of all clinical picture to the corticosteroid therapy.Sarcoid neuropathy can rarely be the presenting feature of sarcoidosis. PMID- 11864562 TI - [Posterior spinal artery infarction and nonbacterial thrombotic endocarditis]. AB - Nonbacterial thrombotic endocarditis (NBTE) has been described as an important complication in patients affected by different kind of illnesses, above all underlying malignancies. Fifty % of the patients affected by NBTE will have embolic events in different organs. The poverty of specific symptomatology appears to difficult seriously its diagnosis. In fact, systemic embolic infarctions are usually asymptomatic, except when central nervous system is involved. We present a patient affected by NBTE with an asymptomatic cervical spinal cord infarction, which diagnosis was made by pathologic study. The difficulty to explore typical sings of spinal lesion and the absence of previous reported cases of NBTE with posterior spinal artery infarction, make this one fairly interesting. We think that fibrinoplatelet embolism from NBTE should be included in the etiology of the posterior spinal artery infarction. PMID- 11864561 TI - [Status cataplecticus induced by abrupt withdrawal of clomipramine]. AB - INTRODUCTION: Cataplexy is one of the main narcoleptic symptoms and is characterized by sudden loss of muscle tone triggered by emotional stimuli while consciousness is mantained. Clomipramine is an effective treatment of cataplexy. Cataplexy that occurs repeatedly for hours or days is referred to as status cataplecticus. PATIENTS: We report three adults with narcolepsy in whom cataplexy was chronically and effectively treated with clomipramine (75-150 mg/day). For diverse reasons, these three patients had an abrupt withdrawal of clomipramine, and after 2-9 days patients showed an invalidant status cataplecticus characterized by a marked increase of the frequency, duration and severity of their cataplectic attacks that were now elicited by mild emotional stimuli. After introduction of anticataplectic agents (clomipramine in two patients and fluoxetine in one patient), status cataplecticus was resolved in less than a week. CONCLUSION: In patients with narcolepsy, abrupt withdrawal of chronic treatment with clomipramine may be associated with status cataplecticus. This condition may be resolved with the reintroduction of anticataplectic agents. PMID- 11864563 TI - [Window-shopping in Parkinsonian patients with festinating gait]. PMID- 11864564 TI - [Chronic activation of T-and B-cells in HTLV-1-associated myelopathy]. PMID- 11864565 TI - Identification of a novel Ras-regulated proapoptotic pathway. AB - BACKGROUND: The Ras-GTPase controls cell fate decisions through the binding of an array of effector molecules, such as Raf and PI 3-kinase, in a GTP-dependent manner. NORE1, a noncatalytic polypeptide, binds specifically to Ras-GTP and to several other Ras-like GTPases. NORE is homologous to the putative tumor suppressor RASSF1 and to the Caenorhabditis elegans polypeptide T24F1.3. RESULTS: We find that all three NORE-related polypeptides bind selectively to the proapoptotic protein kinase MST1, a member of the Group II GC kinases. Endogenous NORE and MST1 occur in a constitutive complex in vivo that associates with endogenous Ras after serum stimulation. Targeting recombinant MST1 to the membrane, either through NORE or myristoylation, augments the apoptotic efficacy of MST1. Overexpression of constitutively active Ki-RasG12V promotes apoptosis in a variety of cell lines; Ha-RasG12V is a much less potent proapoptotic agent; however, a Ha-RasG12V effector loop mutant (E37G) that binds NORE, but not Raf or PI 3-kinase, exhibits proapoptotic efficacy approaching that of Ki-RasG12V. The apoptotic action of both Ki-RasG12V and Ha-RasG12V, E37G is suppressed by overexpression of the MST1 carboxy-terminal noncatalytic segment or by the NORE segment that binds MST1. CONCLUSIONS: MST1 is a phylogenetically conserved partner of the NORE/RASSF polypeptide family, and the NORE-MST1 complex is a novel Ras effector unit that mediates the apoptotic effect of Ki-RasG12V. PMID- 11864566 TI - Multiple Skp1-related proteins in Caenorhabditis elegans: diverse patterns of interaction with Cullins and F-box proteins. AB - BACKGROUND: The ubiquitin-proteasome pathway of proteolysis controls the abundance of specific regulatory proteins. The SCF complex is a type of ubiquitin protein ligase (E3) that contributes to this pathway in many biological systems. In yeast and mammals, the SCF complex consists of common components, including Skp1, Cdc53/Cul1, and Rbx1, as well as variable components known as F-box proteins. Whereas only one functional Skp1 gene is present in the human genome, the genome of Caenorhabditis elegans has now been shown to contain at least 21 Skp1-related (skr) genes. The biochemical properties, expression, and function of the C. elegans SKR proteins were examined. RESULTS: Of the 17 SKR proteins examined, eight (SKR-1, -2, -3, -4, -7, -8, -9, and -10) were shown to interact with C. elegans CUL1 by yeast two-hybrid analysis or a coimmunoprecipitation assay in mammalian cells. Furthermore, SKR proteins exhibited diverse binding specificities for C. elegans F-box proteins. The tissue specificity of expression of the CUL1-interacting SKR proteins was also varied. Suppression of skr-1 or skr 2 genes by double-stranded RNA interference resulted in embryonic death, whereas that of skr-7, -8, -9, or -10 was associated with slow growth and morphological abnormalities. CONCLUSIONS: The multiple C. elegans SKR proteins exhibit marked differences in their association with Cullins and F-box proteins, in tissue specificity of expression, and in phenotypes associated with functional suppression by RNAi. At least eight of the SKR proteins may, like F-box proteins, act as variable components of the SCF complex in C. elegans. PMID- 11864567 TI - The Caenorhabditis elegans Skp1-related gene family: diverse functions in cell proliferation, morphogenesis, and meiosis. AB - BACKGROUND: The SCF ubiquitin-ligase complex targets the ubiquitin-mediated degradation of proteins in multiple dynamic cellular processes. A key SCF component is the Skp1 protein that functions within the complex to link the substrate-recognition subunit to a cullin that in turn binds the ubiquitin conjugating enzyme. In contrast to yeast and humans, Caenorhabditis elegans contains multiple expressed Skp1-related (skr) genes. RESULTS: The 21 Skp1 related (skr) genes in C. elegans form one phylogenetic clade, suggesting that a single ancestral Skp1 gene underwent independent expansion in C. elegans. The cellular and developmental functions of the 21 C. elegans skr genes were probed by dsRNA-mediated gene inactivation (RNAi). The RNAi phenotypes of the skr genes fall into two classes. First, the highly similar skr-7, -8, -9, and -10 genes are required for posterior body morphogenesis, embryonic and larval development, and cell proliferation. Second, the related skr-1 and -2 genes are required for the restraint of cell proliferation, progression through the pachytene stage of meiosis, and the formation of bivalent chromosomes at diakinesis. CUL-1 was found to interact with SKR-1, -2, -3, -7, -8, and -10 in the yeast two-hybrid system. Interestingly, SKR-3 could interact with both CUL-1 and its close paralog CUL-6. CONCLUSIONS: Members of the expanded skr gene family in C. elegans perform critical functions in regulating cell proliferation, meiosis, and morphogenesis. The finding that multiple SKRs are able to bind cullins suggests an extensive set of combinatorial SCF complexes. PMID- 11864568 TI - Multiple levels of posttranscriptional control lead to germ line-specific gene expression in the zebrafish. AB - An important mechanism for the specification and development of the animal germ line is the localization of specific molecules to the germ plasm. Restriction of these molecules to the germ line is considered to be critical for proper development of the germ line as well as the soma. Cytoplasmic localization alone, however, may not be sufficient to achieve germ line-specific expression. While zebrafish vasa mRNA is localized to the germ plasm, the Vasa protein is initially distributed uniformly in the embryo, and its expression becomes restricted to the PGCs only later in development. Here, we demonstrate that, in addition to vasa RNA localization, multiple cell type-specific posttranscriptional mechanisms act on vasa mRNA and Vasa protein. We show that the portion of the maternal vasa mRNA, which is partitioned to somatic cells, is rapidly degraded, whereas vasa RNA is stabilized in the PGCs in a process that is mediated by cis-acting elements within the molecule. Similarly, the Vasa protein is highly unstable in somatic cells, but not in the PGCs. Finally, we demonstrate that subcellular localization of Vasa protein involves cis-acting domains within the protein. In conclusion, we show that posttranscriptional degradation-protection mechanisms acting on RNA and protein function in a vertebrate to enrich for specific molecules in the PGCs. PMID- 11864569 TI - An unexpected role for visual feedback in vehicle steering control. AB - Some motor tasks can be completed, quite literally, with our eyes shut. Most people can touch their nose without looking or reach for an object after only a brief glance at its location. This distinction leads to one of the defining questions of movement control: is information gleaned prior to starting the movement sufficient to complete the task (open loop), or is feedback about the progress of the movement required (closed loop)? One task that has commanded considerable interest in the literature over the years is that of steering a vehicle, in particular lane-correction and lane-changing tasks. Recent work has suggested that this type of task can proceed in a fundamentally open loop manner, with feedback mainly serving to correct minor, accumulating errors. This paper reevaluates the conclusions of these studies by conducting a new set of experiments in a driving simulator. We demonstrate that, in fact, drivers rely on regular visual feedback, even during the well-practiced steering task of lane changing. Without feedback, drivers fail to initiate the return phase of the maneuver, resulting in systematic errors in final heading. The results provide new insight into the control of vehicle heading, suggesting that drivers employ a simple policy of "turn and see," with only limited understanding of the relationship between steering angle and vehicle heading. PMID- 11864570 TI - Direct long-term observation of kinesin processivity at low load. AB - The hand-over-hand stepping mechanism of kinesin at low loads is inadequately understood because the number of molecular steps taken per encounter with the microtubule is difficult to measure: optical traps do not register steps at zero load, while evanescent wave microscopy of single molecules of GFP-kinesin suffers from premature photobleaching. Obtaining low-load data is important because it can efficiently distinguish between alternative proposed mechanisms for molecular walking. We report a novel experiment that records the missing data. We fused kinesin to gelsolin, creating a construct that severs and caps rhodamine phalloidin actin filaments, setting exactly one kinesin molecule on one end of each fluorescent actin filament. Single kinesin molecules labeled in this way can be tracked easily and definitively using a standard epifluorescence microscope. We use the new system to show that, contrary to a recent report, kinesin run length at low load is independent of ATP concentration in the muM to mM range of ATP concentration. Adding competitor ADP in the presence of saturating ATP decreases both velocity and run length. Based on these data, we propose a simplified model for the mechanism of processive stepping. PMID- 11864571 TI - The Human Rho-GEF trio and its target GTPase RhoG are involved in the NGF pathway, leading to neurite outgrowth. AB - Rho-GTPases control a wide range of physiological processes by regulating actin cytoskeleton dynamics. Numerous studies on neuronal cell lines have established that Rac, Cdc42, and RhoG activate neurite extension, while RhoA mediates neurite retraction. Guanine nucleotide exchange factors (GEFs) activate Rho-GTPases by accelerating GDP/GTP exchange. Trio displays two Rho-GEF domains, GEFD1, activating the Rac pathway via RhoG, and GEFD2, acting on RhoA, and contains numerous signaling motifs whose contribution to Trio function has not yet been investigated. Genetic analyses in Drosophila and in Caenorhabditis elegans indicate that Trio is involved in axon guidance and cell motility via a GEFD1 dependent process, suggesting that the activity of its Rho-GEFs is strictly regulated. Here, we show that human Trio induces neurite outgrowth in PC12 cells in a GEFD1-dependent manner. Interestingly, the spectrin repeats and the SH3-1 domain of Trio are essential for GEFD1-mediated neurite outgrowth, revealing an unexpected role for these motifs in Trio function. Moreover, we demonstrate that Trio-induced neurite outgrowth is mediated by the GEFD1-dependent activation of RhoG, previously shown to be part of the NGF (nerve growth factor) pathway. The expression of different Trio mutants interferes with NGF-induced neurite outgrowth, suggesting that Trio may be an upstream regulator of RhoG in this pathway. In addition, we show that Trio protein accumulates under NGF stimulation. Thus, Trio is the first identified Rho-GEF involved in the NGF differentiation signaling. PMID- 11864572 TI - Both carboxy-terminal tails of alpha- and beta-tubulin are essential, but either one will suffice. AB - Microtubules (MTs) are organized into distinct systems essential for cell shape, movement, intracellular transport, and division. Electron crystallographic analyses provide little information about how MTs produce diverse structures and functions, perhaps because they failed to visualize the last 10 residues of the alpha- and the last 18 of the beta-tubulin C-terminal tails (CTTs), which likely play a role in MT diversity. CTTs define conserved, nonallelic isotypes in mammals, are major sites of posttranslational modifications (PTMs), are binding sites for microtubule-associated proteins (MAPs), and determine MT motor processivity. Using mutagenesis and homologous gene replacement in Tetrahymena thermophila, we analyzed mutations, deletions, tail switches, and tail duplications of alpha- and beta-tubulin CTTs. We demonstrate that a tail is required for the essential function of both alpha- and beta-tubulin. However, the two tails are interchangeable, and cells grow normally with either an alpha or a beta tail on both tubulins. In addition, an alpha gene containing a duplicated alpha C terminus rescues a lethal mutant lacking all known posttranslational modification sites on the beta C terminus but cannot rescue deletion of the beta tail. Thus, tubulin tails have a second essential function that is not associated with posttranslational modification. PMID- 11864573 TI - The p21-activated kinase PAK is negatively regulated by POPX1 and POPX2, a pair of serine/threonine phosphatases of the PP2C family. AB - The Rho GTPases are involved in many signaling pathways and cellular functions, including the organization of the actin cytoskeleton, regulation of transcription, cell motility, and cell division. The p21 (Cdc42/Rac)-activated kinase PAK mediates a number of biological effects downstream of these Rho GTPases (reviewed by [1]). The phosphorylation state of mammalian PAK is highly regulated: upon binding of GTPases, PAK is potently activated by autophosphorylation at multiple sites, although the mechanisms of PAK downregulation are not known. We now report two PP2C-like serine/threonine phosphatases (POPX1 and POPX2) that efficiently inactivate PAK. POPX1 was isolated as a binding partner for the PAK interacting guanine nucleotide exchange factor PIX. The dephosphorylating activity of POPX correlates with an ability to block the in vivo effects of active PAK. Consonant with these effects on PAK, POPX can also inhibit actin stress fiber breakdown and morphological changes driven by active Cdc42(V12). The association of the POPX phosphatases with PAK complexes may allow PAK to cycle rapidly between active and inactive states; it represents a unique regulatory component of the signaling pathways of the PAK kinase family. PMID- 11864574 TI - Eco1 is a novel acetyltransferase that can acetylate proteins involved in cohesion. AB - Cohesion between sister chromatids is established during S phase and maintained through G2 phase until it is resolved in anaphase (for review, see [1-3]). In Saccharomyces cerevisiae, a complex consisting of Scc1, Smc1, Smc3, and Scc3 proteins, called "cohesin," mediates the connection between sister chromatids. The evolutionary conserved yeast protein Eco1 is required for establishment of sister chromatid cohesion during S phase but not for its further maintenance during G2 or M phases or for loading the cohesin complex onto DNA. We address the molecular functions of Eco1 with sensitive sequence analytic techniques, including hidden Markov model domain fragment searches. We found a two-domain architecture with an N-terminal C2H2 Zn finger-like domain and an approximately 150 residue C-terminal domain with an apparent acetyl coenzyme A binding motif (http://mendel.imp.univie.ac.at/SEQUENCES/ECO1/). Biochemical tests confirm that Eco1 has the acetyltransferase activity in vitro. In vitro Eco1 acetylates itself and components of the cohesin complex but not histones. Thus, the establishment of cohesion between sister chromatids appears to be regulated, directly or indirectly, by a specific acetyltransferase. PMID- 11864575 TI - Cell polarity signaling in Arabidopsis involves a BFA-sensitive auxin influx pathway. AB - Coordination of cell and tissue polarity commonly involves directional signaling. In the Arabidopsis root epidermis, cell polarity is revealed by basal, root tip oriented, hair outgrowth from hair-forming cells (trichoblasts). The plant hormone auxin displays polar movements and accumulates at maximum concentration in the root tip. The application of polar auxin transport inhibitors evokes changes in trichoblast polarity only at high concentrations and after long-term application. Thus, it remains open whether components of the auxin transport machinery mediate establishment of trichoblast polarity. Here we report that the presumptive auxin influx carrier AUX1 contributes to apical-basal hair cell polarity. AUX1 function is required for polarity changes induced by exogenous application of the auxin 2,4-D, a preferential influx carrier substrate. Similar to aux1 mutants, the vesicle trafficking inhibitor brefeldin A (BFA) interferes with polar hair initiation, and AUX1 function is required for BFA-mediated polarity changes. Consistently, BFA inhibits membrane trafficking of AUX1, trichoblast hyperpolarization induced by 2,4-D, and alters the distal auxin maximum. Our results identify AUX1 as one component of a novel BFA-sensitive auxin transport pathway polarizing cells toward a hormone maximum. PMID- 11864577 TI - Cdc42 facilitates invasion but not the actin-based motility of Shigella. AB - The enteric pathogen Shigella utilizes host-encoded proteins to invade the gastrointestinal tract. Efficient invasion of host cells requires the stimulation of Rho-family GTPases and cytoskeletal alterations by Shigella-encoded IpaC. Following invasion and lysis of the phagosome, Shigella exploits the host's actin based polymerization machinery to assemble an actin tail that serves as the propulsive force required for spreading within and between cells. The Shigella surface protein IcsA stimulates actin-tail formation by recruiting host-encoded N WASP to drive Arp2/3-mediated actin assembly. N-WASP is absolutely required for Shigella motility, but not for Shigella invasion. Although Rho-family GTPases have been implicated in both the invasion and motility of Shigella, the role of Cdc42, an N-WASP activator, in this process has been controversial. In these studies, we have examined the role of Cdc42 in Shigella invasion and actin-based motility using Cdc42-deficient cells. We demonstrate that Cdc42 is required for efficient Shigella invasion but reveal a minor Cdc42-independent pathway that can permit Shigella invasion. However, the actin-based motility of Shigella, as well as vaccinia, proceeds unperturbed in the absence of Cdc42. These data further support the involvement of distinct host-encoded proteins in the steps regulating invasion and intercellular spread of Shigella. PMID- 11864576 TI - Replication-associated repair of adenine:8-oxoguanine mispairs by MYH. AB - Cellular DNA is constantly exposed to the risk of oxidation. 8-oxoguanine (8 oxoG) is one of the major DNA lesions generated by oxidation, which is primarily corrected by base excision repair. When it is not repaired prior to replication, replicative DNA polymerases yield misinsertion of an adenine (A) opposite the 8 oxoG on the template strand, generating an A:8-oxoG mispair. MYH, a mammalian homolog of Escherichia coli MutY, is a DNA glycosylase responsible for initiating base excision repair of such a mispair by excising the adenine opposite 8-oxoG. Here, using an in vivo repair system, we show that DNA replication enhances the repair of the A:8-oxoG mispair. Repair efficiency was lower in MYH-deficient murine cells than in MYH-proficient cells. Transfection of the MYH-deficient cells with a wild-type MYH expression vector increased the efficiency of A:8-oxoG repair, indicating that a significant part of this replication-associated repair depends on MYH. Expression of a mutant MYH in which the PCNA binding motif was disrupted did not increase the repair efficiency, thus suggesting that the interaction between PCNA and MYH is critical for MYH-initiated repair of A:8 oxoG. PMID- 11864578 TI - Desktop boost to anthrax studies. PMID- 11864579 TI - Stoking Ground Zero health worries. PMID- 11864580 TI - Features: Franklin prize for women scientists. PMID- 11864581 TI - Toxin clues to beetle relationships. PMID- 11864582 TI - Intraflagellar transport. PMID- 11864583 TI - Wnt signalling: a moving picture emerges from van gogh. AB - Recent studies on vertebrate homologues of the van gogh/strabismus (vang/stbm) gene, a key player in planar cell polarity signalling in Drosophila, show that vang/stbm is involved in patterning and morphogenesis during vertebrate gastrulation where it modulates two distinct Wnt signals. PMID- 11864584 TI - Epigenetics: the flowers that come in from the cold. AB - Plants can remember periods of cold several weeks later and respond by switching from vegetative to reproductive development. Recent findings shed light on this phenomenon by showing that a gene responsible for keeping this memory encodes a member of the Polycomb group of proteins. PMID- 11864585 TI - Microbiology: breaking down biofilms. AB - Biofilms--adherent bacterial communities embedded in a polymer matrix--are common in nature and can cause persistent human infections that are highly resistant to antibiotics. Recent work provides insight into how these communities develop and function, and offers clues to combating them. PMID- 11864586 TI - Vision: when the clock appears to stop. AB - Eye movements produce a temporary loss of visual sensitivity known as saccadic suppression, and a distortion of space perception known as saccadic compression. A new study has reported a seemingly related phenomenon --chronostasis---in which one's perception of time also undergoes an illusory distortion during rapid movements of the eyes. PMID- 11864587 TI - MicroRNAs: hidden in the genome. AB - Genes for tiny RNAs have been found to be plentiful in the genomes of worms, flies, humans and probably all animals. Some of these microRNAs have been conserved through evolution, and many are expressed only at specific times or places. How they act is just beginning to be understood, but their importance to biology is likely to be great. PMID- 11864588 TI - Polyglutamine disease: acetyltransferases awry. AB - Recent evidence indicates that inhibition of histone acetyltransferases may be a primary cause of cellular pathogenesis in polyglutamine diseases such as Huntington disease; the results raise the possibility that pharmacologic manipulation of protein acetylation levels could be of therapeutic benefit. PMID- 11864589 TI - Cortical plasticity: learning from cortical reorganisation. AB - Neocortical circuits can undergo dynamic rearrangements, not only in response to injury, but also when new skills are acquired. But although training can lead to functional rewiring of the cortex, we are far from being able to reprogram an animal by manipulating its cortical circuitry directly. PMID- 11864590 TI - Transcription: surprising role for an elusive small nuclear RNA. AB - Since 7SK was discovered as an abundant small nuclear RNA in the mid-70s, its role has remained tantalizingly unsolved. Two recent studies have shown that 7SK acts as a negative regulator of the RNA polymerase II elongation factor P-TEFb, and that this activity is influenced by stress response pathways. PMID- 11864591 TI - Direct interactions between rab GTPases and cargo. AB - The rab family of small GTPases has numerous roles in intracellular transport including budding, tethering, and fusion of vesicles as well as organelle motility. New data show that cargo proteins are also rab effectors and can therefore regulate their own trafficking by direct interactions with the transport machinery. PMID- 11864592 TI - FtsK: Maxwell's Demon? AB - FtsK, which links chromosome segregation and cell division in E. coli, has now been shown to be an ATP-dependent DNA translocase. It also activates XerCD dependent recombination, converting chromosome dimers into monomers, by switching the order of strand cleavage by the recombinase subunits. PMID- 11864593 TI - The FEAR factor. AB - The decision to exit from mitosis is a fateful one, and Amon and colleagues have shown that it involves both an early and a late component. The FEAR network initiates the exit machinery, while the previously described MEN pathway maintains it. PMID- 11864594 TI - Opening chromatin. AB - How the transcriptional apparatus gains access to and subsequently opens compacted chromatin has remained a mystery. Now in this issue of Molecular Cell, Zaret and coworkers show that chromatin opening in vitro is not mediated by the classical chromatin remodeling machines but instead occurs through specific transcription factor-histone interactions. PMID- 11864596 TI - Don C. Wiley (1944--2001). PMID- 11864595 TI - CtBP, an unconventional transcriptional corepressor in development and oncogenesis. AB - CtBP family proteins are conserved among vertebrates and invertebrates and function as transcriptional corepressors. They repress transcription in a histone deacetylase-dependent or -independent manner. CtBPs play important roles during development and oncogenesis. In this review, their unusual properties, the mechanisms of transcriptional repression, regulation, and their biological functions are discussed. PMID- 11864597 TI - A unified nomenclature for the superfamily of TRP cation channels. PMID- 11864598 TI - Xenopus Mcm10 binds to origins of DNA replication after Mcm2-7 and stimulates origin binding of Cdc45. AB - Current models suggest that the replication initiation factor Mcm10 is required for association of Mcm2-7 with origins of replication to generate the prereplicative complex (pre-RC). Here we report that Xenopus Mcm10 (XMcm10) is not required for origin binding of XMcm2-7. Instead, the chromatin binding of XMcm10 at the onset of DNA replication requires chromatin-bound XMcm2-7, and it is independent of Cdk2 and Cdc7. In the absence of XMcm10, XCdc45 binding, XRPA binding, and initiation-dependent plasmid supercoiling are blocked. Therefore, XMcm10 performs its function after pre-RC assembly and before origin unwinding. As one of the earliest known pre-RC activation steps, chromatin binding of XMcm10 is an attractive target for regulation by cell cycle checkpoints. PMID- 11864599 TI - Direct rescue of stalled DNA replication forks via the combined action of PriA and RecG helicase activities. AB - The PriA protein of Escherichia coli plays a key role in the rescue of replication forks stalled on the template DNA. One attractive model of rescue relies on homologous recombination to establish a new fork via PriA-mediated loading of the DnaB replicative helicase at D loop intermediates. We provide genetic and biochemical evidence that PriA helicase activity can also rescue a stalled fork by an alternative mechanism that requires manipulation of the fork before loading of DnaB on the lagging strand template. This direct rescue depends on RecG, which unwinds forks and Holliday junctions and interconverts these structures. The combined action of PriA and RecG helicase activities may thus avoid the potential dangers of rescue pathways involving fork breakage and recombination. PMID- 11864600 TI - Meiotic recombination remote from prominent DNA break sites in S. pombe. AB - DNA breakage is intimately associated with meiotic recombination in the fission yeast Schizosaccharomyces pombe. Sites of prominent DNA breakage were found approximately 25 to approximately 200 kb apart in the genomic regions surveyed. We examined in detail a 501 kb region of chromosome I and found six sites, or tight clusters of sites, at which approximately 2%-11% of the DNA accumulated breaks in a rad50S mutant. In contrast to the discrete, widely spaced distribution of prominent break sites, recombination in this region was more uniformly distributed (0.7-1.6 cM/10 kb) whether the genetic interval tested contained no, one, or more such sites. We infer that although recombination depends upon DNA breakage, recombination often occurs remote from these sites (tens of kilobases away); we discuss mechanisms by which this may occur. PMID- 11864601 TI - Association of CBP/p300 acetylase and thymine DNA glycosylase links DNA repair and transcription. AB - DNA repair in chromatin is subject to topological constraints, suggesting a requirement for chromatin modification and remodeling activities. Thymine DNA glycosylase (TDG) initiates repair of G/T and G/U mismatches, commonly associated with CpG islands, by removing thymine and uracil moieties. We report that TDG associates with transcriptional coactivators CBP and p300 and that the resulting complexes are competent for both the excision step of repair and histone acetylation. Furthermore, TDG stimulates CBP transcriptional activity in transfected cells and reciprocally serves as a substrate for CBP/p300 acetylation. Remarkably, this acetylation triggers release of CBP from DNA ternary complexes and also regulates recruitment of repair endonuclease APE. These observations reveal a potential regulatory role for protein acetylation in base mismatch repair and a role for CBP/p300 in maintaining genomic stability. PMID- 11864602 TI - Opening of compacted chromatin by early developmental transcription factors HNF3 (FoxA) and GATA-4. AB - The transcription factors HNF3 (FoxA) and GATA-4 are the earliest known to bind the albumin gene enhancer in liver precursor cells in embryos. To understand how they access sites in silent chromatin, we assembled nucleosome arrays containing albumin enhancer sequences and compacted them with linker histone. HNF3 and GATA 4, but not NF-1, C/EBP, and GAL4-AH, bound their sites in compacted chromatin and opened the local nucleosomal domain in the absence of ATP-dependent enzymes. The ability of HNF3 to open chromatin is mediated by a high affinity DNA binding site and by the C-terminal domain of the protein, which binds histones H3 and H4. Thus, factors that potentiate transcription in development are inherently capable of initiating chromatin opening events. PMID- 11864603 TI - A defined locus control region determinant links chromatin domain acetylation with long-range gene activation. AB - Gene activation in higher eukaryotes is often under the control of regulatory elements quite distant from their target promoters. It is unclear how such long range control is mediated. Here we show that a single determinant of the human growth hormone locus control region (hGH LCR) located 14.5 kb 5prime prime or minute to the hGH-N promoter has a critical, specific, and nonredundant role in facilitating promoter trans factor binding and activating hGH-N transcription. Significantly, this same determinant plays an essential role in establishing a 32 kb acetylated domain that encompasses the entire hGH LCR and the contiguous hGH-N promoter. These data support a model for long-range gene activation via LCR mediated targeting and extensive spreading of core histone acetylation. PMID- 11864605 TI - RNAi related mechanisms affect both transcriptional and posttranscriptional transgene silencing in Drosophila. AB - Two types of transgene silencing were found for the Alcohol dehydrogenase (Adh) transcription unit. Transcriptional gene silencing (TGS) is Polycomb dependent and occurs when Adh is driven by the white eye color gene promoter. Full-length Adh transgenes are silenced posttranscriptionally at high copy number or by a pulsed increase over a threshold. The posttranscriptional gene silencing (PTGS) exhibits molecular hallmarks typical of RNA interference (RNAi), including the production of 21--25 bp length sense and antisense RNAs homologous to the silenced RNA. Mutations in piwi, which belongs to a gene family with members required for RNAi, block PTGS and one aspect of TGS, indicating a connection between the two types of silencing. PMID- 11864604 TI - Structural and functional evidence for ligand-independent transcriptional activation by the estrogen-related receptor 3. AB - The crystal structure of the ligand binding domain (LBD) of the estrogen-related receptor 3 (ERR3) complexed with a steroid receptor coactivator-1 (SRC-1) peptide reveals a transcriptionally active conformation in absence of any ligand. The structure explains why estradiol does not bind ERRs with significant affinity. Docking of the previously reported ERR antagonists, diethylstilbestrol and 4 hydroxytamoxifen, requires structural rearrangements enlarging the ligand binding pocket that can only be accommodated with an antagonist LBD conformation. Mutant receptors in which the ligand binding cavity is filled up by bulkier side chains still interact with SRC-1 in vitro and are transcriptionally active in vivo, but are no longer efficiently inactivated by diethylstilbestrol or 4 hydroxytamoxifen. These results provide structural and functional evidence for ligand-independent transcriptional activation by ERR3. PMID- 11864606 TI - The sigma(70)-like motif: a eukaryotic RNA binding domain unique to a superfamily of proteins required for ribosome biogenesis. AB - Little is understood about the role of nucleolar RNA binding proteins in ribosome biogenesis, although there is a clear need for them based on the strict folding requirements of the pre-rRNA. We have identified a superfamily of RNA binding proteins whose members are required for different stages of ribosome biogenesis. The Imp4 superfamily is composed of five individual families (Imp4, Rpf1, Rpf2, Brx1, and Ssf) that all possess the sigma(70)-like motif, a eukaryotic RNA binding domain with prokaryotic origins. The Imp4 superfamily members associate with RNAs that are consistent with their distinct roles in ribosome biogenesis and suggest the mechanisms by which they function. PMID- 11864607 TI - Ssf1p prevents premature processing of an early pre-60S ribosomal particle. AB - Ssf1p and Ssf2p are two nearly identical and functionally redundant nucleolar proteins. In the absence of Ssf1p and Ssf2p, the 27SA(2) pre-rRNA was prematurely cleaved, inhibiting synthesis of the 27SB and 7S pre-rRNAs and the 5.8S and 25S rRNA components of the large ribosomal subunit. On sucrose gradients, Ssf1p sedimented with pre-60S ribosomal particles. The 27SA(2), 27SA(3), and 27SB pre rRNAs were copurified with tagged Ssf1p, as were 23 large subunit ribosomal proteins and 21 other proteins implicated in ribosome biogenesis. These included four Brix family proteins, Ssf1p, Rpf1p, Rpf2p, and Brx1p, indicating that the entire family functions in ribosome synthesis. This complex is distinct from recently reported pre-60S complexes in RNA and protein composition. We describe a multistep pathway of 60S preribosome maturation. PMID- 11864608 TI - Mutational separation of two pathways for editing by a class I tRNA synthetase. AB - Aminoacyl tRNA synthetases (aaRSs) catalyze the first step in protein biosynthesis, establishing a connection between codons and amino acids. To maintain accuracy, aaRSs have evolved a second active site that eliminates noncognate amino acids. Isoleucyl tRNA synthetase edits valine by two tRNA(Ile) dependent pathways: hydrolysis of valyl adenylate (Val-AMP, pretransfer editing) and hydrolysis of mischarged Val-tRNA(Ile) (posttransfer editing). Not understood is how a single editing site processes two distinct substrates--an adenylate and an aminoacyl tRNA ester. We report here distinct mutations within the center for editing that alter adenylate but not aminoacyl ester hydrolysis, and vice versa. These results are consistent with a molecular model that shows that the single editing active site contains two valyl binding pockets, one specific for each substrate. PMID- 11864609 TI - Tim22, the essential core of the mitochondrial protein insertion complex, forms a voltage-activated and signal-gated channel. AB - The protein insertion complex of the mitochondrial inner membrane is crucial for import of the numerous multitopic membrane proteins with internal targeting signals. Little is known about the molecular mechanism of this complex, including whether it forms a real channel or merely acts as scaffold for protein insertion. We report the unexpected observation that Tim22 is the only essential membrane integrated subunit of the complex. Reconstituted Tim22 forms a hydrophilic, high conductance channel with distinct opening states and pore diameters. The channel is voltage-activated and specifically responds to an internal targeting signal, but not to presequences. Thus, a protein insertion complex can combine three essential functions, signal recognition, channel formation, and energy transduction, in one central component. PMID- 11864610 TI - Structure of the Epstein-Barr virus gp42 protein bound to the MHC class II receptor HLA-DR1. AB - Epstein-Barr virus (EBV) causes infectious mononucleosis, establishes long-term latent infections, and is associated with a variety of human tumors. The EBV gp42 glycoprotein binds MHC class II molecules, playing a critical role in infection of B lymphocytes. EBV gp42 belongs to the C-type lectin superfamily, with homology to NK receptors of the immune system. We report the crystal structure of gp42 bound to the human MHC class II molecule HLA-DR1. The gp42 binds HLA-DR1 using a surface site that is distinct from the canonical lectin and NK receptor ligand binding sites. At the canonical ligand binding site, gp42 forms a large hydrophobic groove, which could interact with other ligands necessary for EBV entry, providing a mechanism for coupling MHC recognition and membrane fusion. PMID- 11864611 TI - Reversible oxidation and inactivation of protein tyrosine phosphatases in vivo. AB - We have investigated the regulation of protein tyrosine phosphatases (PTPs) by reactive oxygen species (ROS) in a cellular environment. We demonstrate that multiple PTPs were reversibly oxidized and inactivated following treatment of Rat 1 cells with H(2)O(2) and that inhibition of PTP function was important for ROS induced mitogenesis. Furthermore, we show transient oxidation of the SH2 domain containing PTP, SHP-2, in response to PDGF that requires association with the PDGFR. Our results indicate that SHP-2 inhibits PDGFR signaling and suggest a mechanism by which autophosphorylation of the PDGFR occurs despite its association with SHP-2. The data suggest that several PTPs may be regulated by oxidation and that characterization of this process may define novel links between specific PTPs and particular signaling pathways in vivo. PMID- 11864612 TI - TNF-induced recruitment and activation of the IKK complex require Cdc37 and Hsp90. AB - The IKK complex, containing two catalytic subunits IKKalpha and IKKbeta and a regulatory subunit NEMO, plays central roles in signal-dependent activation of NF kappaB. We identify Cdc37 and Hsp90 as two additional components of the IKK complex. IKKalpha/IKKbeta/NEMO and Cdc37/Hsp90 form an approximately 900 kDa heterocomplex, which is assembled via direct interactions of Cdc37 with Hsp90 and with the kinase domain of IKKalpha/IKKbeta. Geldanamycin (GA), an antitumor agent that disrupts the formation of this heterocomplex, prevents TNF-induced activation of IKK and NF-kappaB. GA treatment reduces the size of the IKK complex and abolishes TNF-dependent recruitment of the IKK complex to TNF receptor 1 (TNF R1). Therefore, heterocomplex formation with Cdc37/Hsp90 is a prerequisite for TNF-induced activation and trafficking of IKK from the cytoplasm to the membrane. PMID- 11864613 TI - A mechanism of cell survival: sequestration of Fas by the HGF receptor Met. AB - Death receptors such as Fas are present in a variety of organs including liver and play an important role in homeostasis. What prevents these harmful receptors from forming homooligomers, clustering, and initiating the apoptotic pathway is not known. Here, we report the discovery of a cell survival mechanism by which Met, a growth factor receptor tyrosine kinase, directly binds to and sequesters the death receptor Fas in hepatocytes. This interaction prevents Fas self aggregation and Fas ligand binding, thus inhibiting Fas activation and apoptosis. Our results describe a direct link between growth factor tyrosine kinase receptors and death receptors to establish a novel paradigm in growth regulation. PMID- 11864614 TI - Three-dimensional structure of the apoptosome: implications for assembly, procaspase-9 binding, and activation. AB - The apoptosome is an Apaf-1 cytochrome c complex that activates procaspase-9. The three-dimensional structure of the apoptosome has been determined at 27 A resolution, to reveal a wheel-like particle with 7-fold symmetry. Molecular modeling was used to identify the caspase recruitment and WD40 domains within the apoptosome and to infer likely positions of the CED4 homology motif and cytochrome c. This analysis suggests a plausible role for cytochrome c in apoptosome assembly. In a subsequent structure, a noncleavable mutant of procaspase-9 was localized to the central region of the apoptosome. This complex promotes the efficient activation of procaspase-3. Therefore, the cleavage of procaspase-9 is not required to form an active cell death complex. PMID- 11864615 TI - RAC protein directs the complete removal of the 3' external transcribed spacer by the Pac1 nuclease. AB - In Schizosaccharomyces pombe, interdependency in rRNA processing is mediated by a large protein complex (RAC) which contains independent binding sites for each of the transcribed spacers. The RAC complex exhibits no nuclease activity but dramatically alters the efficiency and specificity of the Pac1 nuclease, leading to the complete removal of the 3' ETS. Furthermore, the affinity of RAC protein for mutant 3' ETS correlates closely with in vivo effects on rRNA processing, and changes which disrupt RAC protein binding also inhibit Pac1 nuclease cleavage at the 3' end of the 25S rRNA sequence. The observations indicate that, in the presence of the RAC protein/3' ETS complex, cleavage by the RNase III-like homolog is not restricted to the known intermediate sites but also is directed at the 3' end of the 25S rRNA. PMID- 11864616 TI - The divergent U12-type spliceosome is required for pre-mRNA splicing and is essential for development in Drosophila. AB - A minor class of pre-mRNA introns whose excision requires a spliceosome containing U11, U12, U4atac/U6atac, and U5 snRNPs has been identified in plants, insects, and vertebrates. We have characterized single loci that specify the U6atac and U12 snRNAs of Drosophila melanogaster. P element-mediated disruptions of the U6atac and U12 genes cause lethality during the third instar larval and embryonic stages, respectively, and are rescued by U6atac and U12 transgenes. The P element disruption of U6atac results in excision defects of U12-type introns from several transcripts including an alternative U12-dependent spliced isoform of prospero, a homeodomain protein required for CNS development. Thus, we demonstrate the requirement for the U12 spliceosome in the development of a metazoan organism. PMID- 11864619 TI - In vivo and in vitro glycogenic effects of methionine sulfoximine are different in two inbred strains of mice. AB - We investigated the relationship between brain glycogen anabolism and methionine sulfoximine (MSO)-induced seizures in two inbred mouse strains that presented differential susceptibility to the convulsant. CBA/J was considered a MSO-high reactive strain and C57BL/6J a MSO-low-reactive strain. Accordingly, the dose of MSO needed to induce seizures in CBA/J mice is lower than that in C57BL/6J mice, and CBA/J mice which had seizures, died during the first convulsion. In addition, the time--course of the MSO effect is faster in CBA/J mice than that in C57BL/6J mice. Analyses were performed in C57BL/6J and CBA/J mice after administration of 75 (subconvulsive dose) and 40 mg/kg of MSO (subconvulsive dose, not lethal dose), respectively. In the preconvulsive period, MSO induced an increase in the brain glycogen content of C57BL/6J mice only. Twenty-four hours after MSO administration, the brain glycogen content increased in both strains. The activity and expression of fructose-1,6-bisphosphatase, the last key enzyme of the gluconeogenic pathway, were increased in MSO-treated C57BL/6J mice as compared to control mice, at all experimental time points, whereas they were increased in CBA/J mice only 24 h after MSO administration. These latter results correspond to CBA/J mice that did not have seizures. Interestingly, the differences observed in vivo were consistent with results in primary cultured astrocytes from the two strains. This data suggests that the metabolism impairment, which was not a consequence of seizures, could be related to the difference in seizure susceptibility between the two strains, depending on their genetic background. PMID- 11864620 TI - Synchronous firing dynamics in a heterogeneous neurosecretory-cell population in an insect. AB - Five pairs of neurosecretory cells in the subesophageal ganglion of the silkmoth Bombyx mori discharge action potentials in (near) synchrony to release a pheromone biosynthesis-activating neuropeptide (PBAN). Waveforms of compound action potentials recorded extracellularly from axonal tracts were analyzed to determine the firing activity, timing of spikes and the combination of active cells. Analyses revealed a heterogeneous cellular organization of the neurosecretory cell system. There was a gradient in the firing activity among the cells and the activity of a cell was closely related to relative timing of firing: the most active cell was usually the first to fire and participated in about 90% of all synchronous firing events, while the least active unit was mostly the last to fire and contributed to only 40% of all firing events. A cell with a higher firing activity had a higher potential to mediate propagation of synchronous firing in the cell system. Firing activities of right and left cell groups usually differed and the difference increased in case of a low temperature. Synchronous firings occurred more frequently among the same subgroup of cells rather than different subgroups. Heterogeneous cellular organization and coupling may be important for producing a graded pattern of active cell numbers, which seems to be suitable for maintaining a stable firing (secretory) activity of the cell system for a long period of time. PMID- 11864621 TI - Agonist-specific maturation of GIRK current responses in acutely isolated pyramidal neurons of rat neocortex. AB - We performed whole-cell recordings from acutely isolated pyramidal cell somata of rat neocortex to measure and compare G protein-activated, inwardly rectifying K+ (GIRK) currents induced by adenosine, serotonin and baclofen at different postnatal stages (postnatal days 3-19). In about two thirds of neurons, baclofen induced GIRK currents were already detected at postnatal days 3 and 4 (P3-P4) and almost all neurons between P5 and P19 were responsive. This robust response suggests that postsynaptic effects of baclofen occur much earlier than previously thought. Sensitivity to adenosine was around 70% during the first two postnatal weeks. Given the late maturation of functional synaptic inhibition in neocortex, we propose that phasic and/or tonic activation of GIRK current by baclofen and adenosine might serve as a mechanism to control neuronal excitability during early postnatal development. In marked contrast to the pronounced early sensitivity to baclofen and adenosine, only 20% of the neurons displayed a GIRK current response to serotonin during the first postnatal week. After that, about half of the neurons tested positive for serotonin. GIRK current densities for baclofen and adenosine attained a maximum at the end of the second postnatal week, whereas the serotonin-induced current showed a linear increase during the second and third week of life. Set in relationship with previous data on the postnatal expression of receptor protein and GIRK channel mRNA, our findings suggest that the maturation of GIRK current responses is determined predominantly by the different postnatal patterns of receptor expression. PMID- 11864622 TI - Dysregulation of hypothalamic serotonin turnover in diet-induced obese rats. AB - When outbred Sprague--Dawley rats are placed on a diet relatively high in fat and calories (HE diet), half develop diet-induced obesity (DIO), while the rest are diet-resistant (DR). When fed a low fat chow diet from weaning, DIO- and DR-prone rats weigh the same, but DIO-prone rats have a number of abnormalities of neural function, many of which are normalized when they become obese after chronic exposure to a HE diet. Because of its important role in the regulation of energy homeostasis in the hypothalamus, we examined the ratio of serotonin (5-HT) to its metabolite, 5-hydroxyindolacetic acid (5HIAA), as an index of transmitter turnover in micropunches from various brain areas in these rats. While still on chow, both DIO- and DR-prone rats showed lower 5-HT turnover in most brain areas sampled during the last hour of the light phase, when animals become active and begin foraging for food, as compared to the first hour of the light phase, when animals are generally quiescent and not eating. However, unlike DR-prone rats, DIO-prone rats did not show a significant time-dependent difference in 5-HT turnover in either the arcuate or paraventricular hypothalamic nuclei. When fasted for 48 h, both DIO- and DR-prone rats showed a generalized 16--46% decrease in 5-HT turnover in the dorsomedial nucleus, perifornical lateral hypothalamus, dentate gyrus and motor cortex as compared to their free-fed counterparts. However, fasted DIO-prone rats showed a 53% greater reduction in the ventromedial nucleus turnover than fasted DR-prone rats. Finally, when DIO rats became obese after 14 weeks on the HE diet, their abnormalities in hypothalamic 5-HT turnover at the end of the light phase were normalized. Thus, DIO-prone rats show abnormalities of diurnal and fasting-induced alterations in brain 5-HT turnover which may predispose them to become obese when dietary fat and caloric density are increased. Once obesity develops, these abnormalities, like those of several other hypothalamic transmitters and peptides, are normalized. This may contribute to the persistence of obesity once it develops. PMID- 11864623 TI - Lateralisation of projections from the rostral ventrolateral medulla to sympathetic preganglionic neurons in the rat. AB - Spinally projecting sympathoexcitatory neurons in the rostral ventrolateral medulla (RVLM), synapse with sympathetic preganglionic neurons (SPN) and regulate the activity of sympathetic nerves that control the heart, blood pressure and the adrenal medulla (AM). However, the degree of lateralization of the bulbospinal projections to SPN innervating specific targets is poorly understood. Three approaches were used in this study. Anterograde tracer was iontophoresed into a pressor site in the RVLM (left or right) and retrograde tracer injected into the superior cervical ganglion (SCG, right) and the AM (left). Close appositions between anterogradely labelled axons and retrogradely labelled SCG- or AM-SPN were counted. Projections to the SCG were bilateral. Projections to the AM were markedly ipsilateral. In the second part, retrograde tracers were injected unilaterally into the region of the intermediolateral cell column at spinal segment T2 or T8 on one side and the number of labelled neurons in the RVLM counted. The results from each level of injection were similar showing that approximately 63-64% of the neurons were ipsilateral. Responses to glutamate microinjection into the RVLM on adrenal nerve (left) and superior cervical nerve (left) activity were measured. The ratio of the nerve responses was the same even when different sides of the RVLM were injected. The anterograde data strongly suggest that the RVLM projections to AM-SPN are predominantly ipsilateral. Although other experimental approaches also attempted to investigate lateralization, the retrograde data target different and functionally heterogeneous pools of SPN that may mask the ipsilateral projection to the AM. Similarly, chemical stimulation of the RVLM will excite not only monosynaptic projections but also polysynaptic projections that may also mask the predominant ipsilateral monosynaptic projection to AM. PMID- 11864624 TI - Post-hatching hormonal modulation of a sexually dimorphic neuromuscular system controlling song in zebra finches. AB - Sexual dimorphisms are present throughout the zebra finch song system, from forebrain centers to the tracheosyringeal portion of the hypoglossal nucleus (nXIIts) to the muscles of the syrinx (vocal organ). In females, gonadal steroids administered during development can partially masculinize the telencephalic areas, and in adulthood can increase the size of syrinx muscles. In the present study, two experiments were designed to investigate the role of early androgen and estrogen in the development of nXIIts and the ventralis and dorsalis muscles of the syrinx. In experiment one, males and females were treated with testosterone, estradiol, dihydrotestosterone, the anti-androgen flutamide, or a vehicle control for 21 days after hatching. At day 60, nXIIts volume, motoneuron soma size and number were assessed, as well as syrinx weight and the size of ventralis and dorsalis fibers. In experiment two, animals were administered either the estrogen synthesis inhibitor, fadrozole, or vehicle, and the syrinx measurements were taken at day 60. Male-biased sex differences were detected on all measures in both experiments, and several right-side biases were detected. In females, dihydrotestosterone masculinized soma size in nXIIts and testosterone slightly increased syrinx weight. E2 feminized the syrinx of males. However, flutamide did not prevent masculine development of either structure, and fadrozole did not inhibit feminine syrinx development. These results are consistent with the idea that, as in the forebrain, steroid hormones can stimulate aspects of sexual differentiation, but they may not be direct triggers for the process. PMID- 11864625 TI - Glial transport of the neuromodulator D-serine. AB - D-Serine is an endogenous agonist of NMDA receptors that occurs in astrocytes in gray matter areas of the brain. D-Serine is synthesized from L-serine by the activity of a glial enriched serine racemase, but little is known on the properties of D-serine transport and factors regulating its synaptic concentration. In the present report we characterize the transport of D-serine in astrocytes. In primary astrocyte cultures, D-serine uptake is dependent on sodium ions and exhibits both low affinity and low specificity for D-serine. The kinetics of D-serine transport resembles that of ASCT type transporters as several small neutral amino acids strongly inhibit the uptake of D-serine. D Serine fluxes are coupled to counter-movement of L-serine and to a less extent to other small neutral amino acids. Thus, addition of D-serine to cell cultures elicits robust efflux of intracellular L-serine. Conversely, physiological concentrations of L-serine induce efflux of preloaded D-serine from astrocytes. L Serine was more effective than kainate, which have been previously shown to induce D-serine release from astrocytes upon stimulation of non-NMDA type of glutamate receptors. The features of D-serine transport we describe reveal possible new mechanisms controlling the synaptic concentration of D-serine. PMID- 11864626 TI - Coordination between the masticatory and tongue muscles as seen with different foods in consistency and in reflex activities during natural chewing. AB - To study the coordination between the masticatory and extrinsic tongue muscles during natural chewing, electromyographic activities in the digastric (Dig) as a jaw opener, the masseter (Mas) as a jaw closer, the genioglossus (Gg) as a tongue protruder, and the styloglossus (Sg) as a tongue retractor as well as jaw movement trajectories were recorded while rabbits chewed soft, hard, and very hard foods. The Dig and Gg were active in the jaw-opening phase (OP active group), and the Mas and Sg were active in the jaw-closing phase (CL active group). Food consistency affected differently on the duration of burst activities between the muscle groups, i.e. in the CL active group, the duration was longer for the harder food, while there was no difference in the duration of the OP active group among the foods. During hard food chewing in particular, we confirmed our recent findings that reflexly-induced short but large bursts of activity could be documented in the Dig during the jaw-closing phase. Similar short bursts were also documented in the Gg as with the Dig in this study. Inhibitory periods were often observed in the Mas with the Dig short burst and were also observed in the Sg along with the Gg short burst; however the inhibitory effect in the Sg was less pronounced. These findings suggest that: (1) both masticatory and extrinsic tongue muscles are active in a well-coordinated manner during stable chewing, but that (2) reflex effects on antagonistic muscles (i.e. Dig vs. Mas in the masticatory muscles, Gg vs. Sg in the tongue muscles) evoked by tooth contact during chewing may not be analogous between the two muscle groups. PMID- 11864627 TI - IL-2/15 receptor-beta gene deletion alters neurobehavioral performance. AB - The common IL-2/15 receptor-beta (IL-2/15Rbeta) is an essential signaling subunit that is shared exclusively by IL-2 and IL-15, and is enriched in the hippocampal formation and related limbic regions. We have previously shown that mice lacking IL-2 exhibit alterations in hippocampal-dependent learning, sensorimotor gating and accompanying reductions in hippocampal infrapyramidal mossy neuronal fiber length. Although the effects of exogenous IL-2 on various aspects of forebrain neuronal function are well documented, it is unclear whether IL-15 has neuromodulatory actions. Here we sought to test the hypothesis that the combined loss of the ability of IL-2 and IL-15 to signal through IL-2/15Rbeta in the brain would influence neurobehavioral performance, in particular spatial learning and memory performance. To test this hypothesis, we compared several different domains of behavior in mice that had one or both IL-2/15Rbeta gene alleles deleted. Compared with C57BL/6-IL-2/15Rbeta+/+ wild-type and C57BL/6-IL 2/15Rbeta+/- heterozygote littermates, C57BL/6-IL-2/15Rbeta-/- knockout mice exhibited a deficit in prepulse inhibition of the acoustic startle reflex (PPI). The IL-2/15Rbeta knockout mice also showed significant reductions in acoustic startle reactivity, and modest differences in behavior in the elevated plus-maze test indicative of reduced levels of fearfulness in response to novelty. The IL 2/15Rbeta knockout mice did not differ in locomotor activity in either the plus maze or the Morris water-maze, and contrary to our working hypothesis, they did not differ in spatial learning or memory performance in the water-maze. Further studies are required to determine if these behavioral alterations may be attributable to factors such as the loss of the ability of IL-15 and/or IL-2 to modulate limbic neurons, autoimmunity or genetic factors associated with IL 2/15Rbeta gene deletion. PMID- 11864628 TI - An examination of nucleus accumbens cell firing during extinction and reinstatement of water reinforcement behavior in rats. AB - Electrophysiological recording procedures were used to examine nucleus accumbens (Acb) cell firing in rats (n = 13) during water reinforcement sessions consisting of three phases. During phase one (maintenance), a lever press resulted in water reinforcement (fixed ratio 1; 0.05 ml/press) paired with an auditory stimulus (0.5 s). Of 128 Acb neurons recorded during maintenance, 40 cells (31%) exhibited one of three types of neuronal firing patterns described previously [J. Neurosci. 14 (12) (1994) 7735-7746; J. Neurosci. 20 (11) (2000) 4255-4266]. Briefly, Acb neurons exhibited increases in firing rate within seconds preceding the reinforced response (type PR) or increases (type RFe) or decreases (type RFi) in activity seconds following response completion. In phase two (extinction), subsequent lever pressing had no programmed consequences (i.e., water reinforcement and the auditory stimulus were not presented). After 30 min of no responding, animals were given water reinforcement/auditory stimulus 'primes' to reestablish lever pressing behavior during the third phase (reinstatement). Results indicated that all types of phasic neurons (PR, RFe and RFi) exhibited an attenuated firing rate during extinction, and in some cases recovery of patterned discharges were observed during reinstatement. No significant changes in cell firing were observed for any cell type during presentation of the stimulus prime used to reestablish operant responding following extinction. These findings are discussed in terms of how Acb neurons process information related to 'natural' reinforcers versus drugs of abuse. PMID- 11864629 TI - Loss of mRNA levels, binding and activation of GTP-binding proteins for cannabinoid CB1 receptors in the basal ganglia of a transgenic model of Huntington's disease. AB - Data obtained from the basal ganglia of postmortem Huntington's disease (HD) brains have revealed that the level of cannabinoid CB1 receptors in striatal efferent neurons decreases in parallel to the dysfunction and subsequent degeneration of these neurons. These findings, and others from rat models of HD generated by lesions with mitochondrial toxins, suggest that the loss of CB1 receptors may be involved in the pathogenesis of the disease. To explore further the changes in the endocannabinoid system, as well as the potential of endocannabinoid-related compounds, we examined the status of CB1 receptors in the HD94 transgenic mouse model of HD. These mice express huntingtin exon 1 with a polyglutamine tract of 94 repeats in a tissue-specific and conditional manner using the tet regulatable system. They develop many features of HD, such as striatal atrophy, intraneuronal aggregates and progressive dystonia. In these animals, we analyzed mRNA levels for the CB1 receptor, in addition to the number of specific binding sites and the activation of GTP-binding proteins by CB1 receptor agonists. mRNA transcripts of the CB1 receptor were significantly decreased in the caudate-putamen of HD transgenic mice compared to age-matched littermate controls. The decrease concurred with a marked reduction in receptor density in both the caudate-putamen and its projection areas such as the globus pallidus, entopeduncular nucleus and substantia nigra pars reticulata. Furthermore, the efficacy of CB1 receptor activation was reduced in the globus pallidus, as determined by agonist-induced [35S]GTPgammaS binding, and tended towards a decrease in the substantia nigra. None of these changes was seen in the cerebral cortex and hippocampus, despite high levels of expression of the mutant protein in these regions. The decrease in CB1 receptor levels was accompanied by a decrease in the proenkephalin-mRNA levels but not in substance P-mRNA levels. Taken together, these results suggest that the loss of CB1 receptor might be preferential to the enkephalinergic CB1 receptor-containing striatopallidal neurons, and further implicate the CB1 receptor to the subsequent HD symptomatology and neuropathology. PMID- 11864630 TI - Calbindin D-28k positive projection neurones and calretinin positive interneurones of the rat globus pallidus. AB - Immunohistochemistry for three calcium-binding proteins calbindin D-28k, calretinin, and parvalbumin revealed neuronal heterogeneity within the GP. Each neurone appeared to express either a single type of calcium binding protein or none at all. The co-localisation of calcium binding proteins was not observed. Combined immunohistochemistry and retrograde tract tracing using colloidal gold particles injected into the projection fields, the substantia nigra or subthalamic nucleus, revealed that projection neurones could be labelled with either calbindin or parvalbumin. These cells were of medium size (22 x 12 microm), multipolar and moderate varicose dendritic trees. In contrast, calretinin-positive neurones were never retrogradely labelled, even in regions where neuronal colloidal gold deposits were numerous. This, combined with their rarity (<1%) and small size (11 x 9 microm), suggests that calretinin may be a neurochemical marker for putative rat globus pallidus interneurones. Calcium binding proteins are known to have unique buffering characteristics that may confer specific functional properties upon pallidal neurones. Indeed, differential calcium binding protein expression may underlie the electrophysiological heterogeneity observed in the rat globus pallidus. PMID- 11864631 TI - Neurodegeneration in the rat hippocampus and striatum after middle cerebral artery occlusion. AB - Animal models of ischemia are in wide use to elucidate the molecular mechanisms of brain injury that result from cardiovascular disease in humans. We have used the fluorescent, anionic dye, Fluoro-Jade, to examine cellular degeneration that occurs in association with the middle cerebral artery occlusion (MCAO) model. MCAO results in cortical infarction as well as damage to the hippocampus leading to a delayed form of death of hippocampal neurons. We examined brain sections at 6 h, 12 h, 1, 4, 7, 14 and 21 days after injury. Fluoro-Jade labeling of the striatum was seen over a protracted time-course, with degeneration beginning by 6 h after injury. Neuronal degeneration in the hippocampus, in contrast, occurs between 12 h and 7 days after injury with neuronal death reaching a peak at 4 days. GFAP/Fluoro-Jade double labeling revealed that the Fluoro-Jade positive staining at late time-points in the striatum included astrocytic cells. Together, the results show Fluoro-Jade to be a useful marker of cellular degeneration following ischemic injury. Further, the use of this dye has enabled us to demonstrate previously undescribed events of cellular injury resulting from ischemia. PMID- 11864632 TI - Pharmacological characterisation of the beta-adrenoceptor expressed by human lung mast cells. AB - The nonselective beta-adrenoceptor agonist, isoprenaline (pD2; 8.8 +/- 0.2), and selective beta2-adrenoceptor agonists, clenbuterol (9.2 +/- 0.4) and salbutamol (7.1 +/- 0.1), inhibited the immunoglobulin E-mediated release of histamine from human lung mast cells in a concentration-dependent manner. The beta2-adrenoceptor selective antagonist, ICI118551 (erythro-(+/- )-1-(7-methylindan-4-yloyl)-3 isopropylaminobutan-2-ol HCl), antagonised the isoprenaline inhibition of histamine release from human lung mast cells with high affinity (apparent pK(B); 9.5 +/- 0.2), whereas high concentrations of the beta1-adrenoceptor-selective antagonist, CGP20712A (2-hydroxy-5-(2-(hydroxy-3-(4((1-methyl-4-trifluoromethyl) 1-H-imidazol-2-yl)-phenoxy)-propyl)-aminoethoxyl)-benzamide), were required to reverse the isoprenaline inhibition (apparent pK(B); 6.5 +/- 0.3). Radioligand binding studies using [125I]-iodocyanopindolol ([125I]CYP) were performed on membranes derived from purified mast cells (>90% purity). Binding of [125I]CYP to mast cell membranes was displaced from a single binding site with a high affinity for ICI118551 (pK(i); 8.9 +/- 0.1) and low affinity for CGP20712A (pK(i); 6.0 +/- 0.03), indicative of a homogeneous population of beta2-adrenoceptors. In contrast, in human lung membranes, these antagonists displaced [125I]CYP from two sites indicative of a heterogeneous population of beta1-adrenoceptors (20%) and beta2-adrenoceptors (80%). These data indicate that the beta-adrenoceptor expressed by human lung mast cells and mediating inhibition of mediator release from these cells is the beta2-adrenoceptor. PMID- 11864633 TI - Budesonide reduces multidrug resistance-associated protein 1 expression in an airway epithelial cell line (Calu-1). AB - The objective of this study was to determine the expression and activity of multidrug resistance-associated protein (MRP1) in a human airway epithelial cell line (Calu-1) and to further assess whether budesonide, a potent antiasthma corticosteroid, alters the expression and activity of MRP1 in these cells. Reverse transcriptase polymerase chain reaction (RT-PCR) and the Western blot analysis demonstrated the MRP1 mRNA and MRP1 protein in Calu-1 cells. Indomethacin, probenecid, and verapamil significantly enhanced the fluorescein accumulation and reduced the fluorescein efflux, consistent with the MRP1 activity in the Calu-1 cells. Following 14-day budesonide treatment, fluorescein accumulation increased and fluorescein efflux decreased, consistent with the inhibition of MRP1 activity by budesonide. At a concentration (10 microM) devoid of cytotoxicity, budesonide treatment decreased MRP1 mRNA and MRP1 protein expression in Calu-1 cells by 38% and 42%, respectively. In addition, budesonide (10 microM) enhanced the sensitivity of the MRP1 overexpressing COR-L23R cells to vincristine, suggesting the chemosensitizing effect of budesonide. Thus, budesonide inhibits MRP1 expression and may be useful as a chemosensitizer in tumor chemotherapy. PMID- 11864634 TI - Effects of amifostine on the proliferation and differentiation of megakaryocytic progenitor cells. AB - This study investigated the effects of amifostine, a clinically usable radioprotector or chemoprotector, on the proliferation and differentiation of normal and X-irradiated cluster of differentiation 34 positive (CD34+) megakaryocytic progenitor cells (colony-forming unit in megakaryocytes, CFU-Meg) from human placental and umbilical cord blood (CB) in vitro. Amifostine significantly accelerated megakaryocyte colony formation in a plasma clot culture supplemented with recombinant human thrombopoietin because of an increase in immature CFU-Meg-derived large megakaryocyte colony formation. An analysis of the cells that were harvested from the culture showed that amifostine induced a 70- and an 83-fold increase in the total cell and CFU-Meg numbers, respectively, and produced hyperploid megakaryocytes of more than 8 N ploidy. The radioprotective effect of amifostine on the clonal growth of X-irradiated CD34+ CFU-Meg was observed by treatment before or after irradiation. These findings suggest that the action of amifostine extends from immature CFU-Meg to the terminal differentiation of megakaryopoiesis, and its radioprotective effect is shown in megakaryopoiesis and thrombopoiesis. PMID- 11864635 TI - The human tachykinin NK1 (short form) and tachykinin NK4 receptor: a reappraisal. AB - Excessive secretion of placental neurokinin B into the circulation during the third trimester of pregnancy is seen in women with preeclampsia. To determine a role for neurokinin B, we have used a number of different animal models to ascertain the expression of the three tachykinin receptors (NK1--both short and long forms, NK2 and NK3) and the putative human tachykinin NK4 receptor in the placenta. Human and rat placenta express all three classical tachykinin receptors. However, we failed to reveal the expression of the short tachykinin NK1 receptor or the tachykinin NK4 receptor in any of 24 human tissues examined including the placenta. We conclude that the proposed short form of the tachykinin NK1 receptor is a truncated genomic clone and that the human tachykinin NK4 receptor is in fact, the guinea pig tachykinin NK3 receptor. PMID- 11864636 TI - Mechanism of alpha-subunit selectivity of benzodiazepine pharmacology at gamma aminobutyric acid type A receptors. AB - Benzodiazepine pharmacology at the GABA(A) receptor is dependent on the alpha and gamma subunit isoforms present. Ligands with higher affinity for certain isoforms -selective compounds--have been classified into benzodiazepine type I and II and into diazepam-sensitive and diazepam-insensitive receptors. A single amino acid position (alpha1G201/alpha3E225) has been identified which discriminates BZI and BZII receptors. The role of this residue has been explored by mutagenesis of alpha1 position 201 and the pharmacology of recombinant receptors examined using BZI receptor agonists. Ligand affinity is reduced by increasing side chain volume at alpha1G201 suggesting that steric inhibition underlies alpha-subunit selectivity. A second amino acid (alpha1H102/alpha6R100) determines diazepam sensitivity. The nature of the amino acid at this position was also examined by mutagenesis. Flumazenil and Ro15-4513 (ethyl 8-azido-6-dihydro-5-methyl-6-oxo-4H imidazo[1,5-a]-[1,4]benzodiazepine-3-carboxylate) binding affinity correlated weakly with the amino acid hydrophobicity suggesting a weak hydrophobic interaction between the ligand and alpha1H102. PMID- 11864638 TI - Nitric oxide and convulsions in 4-aminopyridine-treated mice. AB - We studied whether N(G)-nitro-L-arginine (NNA), an inhibitor of nitric oxide (NO) synthase as well as L-arginine and molsidomine, two agents elevating NO, influenced convulsions caused by 4-aminopyridine, a K+ channel blocker in mice. NNA, in a dose known to decrease level of NO (40 mg x kg(-1)), enhanced the seizure susceptibility to intraperitoneal (i.p.) and intracerebroventricular (i.c.v.) 4-aminopyridine. L-arginine (500 mg x kg(-1)) and molsidomine (20 mg x kg(-1)) alone did not influence 4-aminopyridine-induced seizure activity. Surprisingly, the proconvulsant effect of NNA upon clonic and tonic seizures was potentiated by molsidomine (20 mg x kg(-1)). No influence of L-arginine on the proconvulsant effect of NNA was found. Taking into account the proconvulsant effect of NNA, an involvement of NO-mediated events in the mechanism of convulsive activity of 4-aminopyridine might be postulated. However, the ineffectiveness of L-arginine and molsidomine to suppress the convulsive activity of 4-aminopyridine as well as a paradoxical potentiation of the proconvulsant effect of NNA by molsidomine seem to exclude the impact of NO pathway on 4 aminopyridine-induced convulsions in mice. Our data suggest that the proconvulsant effect of NNA in this seizure model is caused by other, not related to NO, mechanisms. PMID- 11864637 TI - Phosphorylation of Ser360 in the third intracellular loop of the alpha2A adrenoceptor during protein kinase C-mediated desensitization. AB - The alpha2A-adrenoceptor undergoes desensitization in response to protein kinase C activation. Using mutagenesis and recombinant expression we sought to determine the specific sites within the receptor which are phosphorylated by protein kinase C and are responsible for this desensitization. Ser/Thr in the third intracellular loop were substituted with Ala to create mutant receptors T272A, S258A, S324A and S360A. These mutations had no effect on ligand binding or functional coupling to inositol phosphate accumulation and intracellular Ca2+ release. Three of the four mutant receptors displayed wild-type desensitization (approximately 60% loss of function) in response to 0.1 microM phorbol-12 myristate-13-acetate (PMA) exposure for 2 min. However, the S360A mutant had only approximately 24% desensitization. In whole cell phosphorylation studies, S360A failed to undergo detectable PMA promoted phosphorylation. We conclude that protein kinase C-mediated desensitization of alpha2A-adrenoceptor function is primarily due to phosphorylation of Ser at amino acid 360. This thus represents one mechanism by which these receptors undergo regulation by heterologous means, such as pathologic processes which activate protein kinase C or crosstalk with other receptors. PMID- 11864639 TI - Effects of acute acamprosate and homotaurine on ethanol intake and ethanol stimulated mesolimbic dopamine release. AB - The purpose of the present study was to determine the acute effects of the anticraving compound acamprosate (calcium acetylhomotaurinate) and the closely related compound homotaurine on ethanol intake and ethanol-stimulated dopamine release in the nucleus accumbens. Male rats were treated with acamprosate (200 or 400 mg/kg intraperitoneally, i.p.) or homotaurine (10, 50, or 100 mg/kg i.p.) 15 min prior to access to 10% ethanol and water for 1 h in a two-bottle choice restricted access paradigm. A separate group of rats was implanted with microdialysis probes in the nucleus accumbens and given an acute injection of ethanol (1.5 g/kg i.p.) that was preceded by saline, acamprosate, or homotaurine. Acamprosate and homotaurine dose-dependently reduced ethanol intake and preference. These compounds also delayed or suppressed ethanol-stimulated increases in nucleus accumbens dopamine release, suggesting that acamprosate and homotaurine may reduce ethanol intake by interfering with the ability of ethanol to activate the mesolimbic dopamine reward system. PMID- 11864640 TI - Spinal transection increases the potency of clonidine on the tail-flick and hindlimb flexion reflexes. AB - The effect of intrathecal clonidine on thermal nociception and hindlimb flexion was assessed in acute and chronic spinally transected rats. After an acute, 1-day spinalization, there was no change in the antinociceptive dose-response function to clonidine, relative to intact rats. However, there was a significant increase in potency 31 days after spinalization. Low doses of clonidine (0.25, 1, 4 and 20 microg) did not affect the nonnociceptive flexion reflex of acute spinal rats, but they elicited a dose-dependent response in chronic spinal rats. These data provide behavioral evidence of supersensitivity to alpha-adrenoceptor agonists in chronic spinal rats. PMID- 11864641 TI - Role of K+ channels in the PACAP-induced catecholamine secretion from the rat adrenal gland. AB - We eluciated whether K+ channels modulate adrenal catecholamine secretion induced by pituitary adenylate cyclase-activating polypeptide (PACAP) in the isolated perfused rat adrenal gland. PACAP (100 nM) increased adrenal epinephrine output. The PACAP-induced responses were enhanced by treatment with apamin (10-100 nM) in a concentration-dependent manner. In the presence of nifedipine (3 microM), apamin (1 microM) did not enhance the PACAP-induced responses. Charybdotoxin (1 100 nM) had little influence on the PACAP-induced responses. These results suggest that small-conductance Ca2+-activated K+ channels interfere with L-type voltage-dependent Ca2+ channels to counteract the PACAP-induced adrenal catecholamine secretion. PMID- 11864642 TI - Impaired visual memory in rats reared in isolation is reversed by D-cycloserine in the adult rat. AB - Previous studies have shown that environmental factors can influence cholinergic and glutamatergic activity in the developing brain, and that the variations in neurochemistry are accompanied by behavioral changes in later life. Rats reared in isolated, social, or enriched environments were tested with a visual discrimination task in adulthood. The results show that saline-treated rats reared in isolation exhibited impaired retention of the discrimination task compared to rats raised in social or enriched environments. However, systemic administration of the NMDA receptor agonist, D-cycloserine (3 mg/kg), restored normal memory function in cognitively impoverished rats. Acquisition of the task was not affected by the rearing conditions. D-Cycloserine is considered to be an efficient cognitive enhancer probably able to compensate for assumed loss of NMDA receptors during isolated rearing. PMID- 11864643 TI - DOI, a 5-HT2 receptor agonist, induces renal vasodilation via nitric oxide in anesthetized dogs. AB - We have previously reported that (+/-)-1-(2.5-dimethoxy-4-iodophenyl)-2 aminopropane (DOI), a 5-HT2 receptor agonist, induced renal vasodilation in anesthetized dogs. The present study was designed to investigate whether DOI induced renal vasodilation might be mediated by increased nitric oxide (NO) release/production in renal tissue. The experiments were performed in anesthetized dogs. A 23-gauge needle was inserted into the left renal artery for infusion of drug solutions. Renal blood flow was measured with an electromagnetic flowmeter. The microdialysis probes were implanted into the renal cortex to collect the dialysate for measurement of guanosine 3',5'-cyclic monophosphate (cGMP) and nitrite/nitrate (NO2/NO3) concentration. Intrarenal infusion of DOI at a rate of 5 microg/kg/min resulted in a significant increase, by 30 +/- 4%, in renal blood flow, indicating renal vasodilation. The renal interstitial concentrations of NO2/NO3 and cGMP also increased by 70 +/- 6% and 60 +/- 6%, respectively. These changes induced by DOI were completely abolished by the intrarenal pretreatment with N(w)-nitro-L-arginine methyl ester (L-NAME, a NO synthase inhibitor, 100 microg/kg/min) or sarpogrelate (100 microg/kg/min, a highly selective 5-HT2 receptor antagonist). DOI infusion increased urine volume and urinary excretion of Na+, which were also blocked by L-NAME or sarpogrelate. These results suggest that DOI caused renal vasodilation due to increased NO release/production by stimulation of 5-HT2 receptors in the kidney. The natriuretic effect of DOI might also be related to increased intrarenal NO production. PMID- 11864644 TI - Amiloride inhibits proximal tubular reabsorption in conscious euvolemic rats. AB - Based on the results of micropuncture studies, it is generally assumed that amiloride inhibits Na+ (and Li+) reabsorption in the distal nephron, without affecting proximal tubular reabsorption. This is the basis for the use of amiloride to test for distal nephron Li+ reabsorption. We have examined the validity of this assumption by administering amiloride in doses of 0, 0.02, 0.07, 0.2 and 2.0 mg x kg(-1) x h(-1) to conscious, chronically instrumented rats fed a diet with a normal Na+ and K+ content. Na+ and water homeostasis was maintained by servo-controlled replacement in order to avoid any effect of volume depletion on proximal tubular reabsorption. The effects of the two highest doses of amiloride were also examined without Na+ and water replacement. In the servo controlled rats, the two highest doses of amiloride increased the fractional excretion of both Na+ (FE(Na)) and Li+ (FE(Li)), whereas the two lowest doses affected only FE(Na). In the rats without servo-control, FE(Li) also rose in response to amiloride infusion, but the increase was significantly lower than that observed in the servo-controlled animals. Since distal Li+ reabsorption is absent or negligible in rats fed a diet with a normal Na+ and K+ content, the large increase in FE(Li) following the highest doses of amiloride (15-18% of the filtered load in servo-controlled rats) indicates inhibition of proximal tubular reabsorption. We conclude that amiloride, in doses usually employed to detect distal Li+ reabsorption, inhibits proximal tubular reabsorption in conscious euvolemic rats. PMID- 11864645 TI - Protective effect of silymarin in antigen challenge- and histamine-induced bronchoconstriction in in vivo guinea-pigs. AB - The effects of silymarin on bronchoconstriction induced by antigen challenge and on post-antigen challenge hyperresponsiveness to substance P were evaluated in sensitized guinea-pigs. Silymarin significantly decreased the bronchoconstriction due to antigen administration in the early phase of the response. In contrast, the dose-response curve for substance P recorded 1 h after antigen challenge was not modified by pretreatment with silymarin. The influence of the flavonoid on hyperresponsiveness to histamine in propranolol- and PAF (platelet-activating factor)-treated animals was also assessed. Silymarin did not affect hyperresponsiveness to histamine induced by either propranolol or PAF although it had inhibitory activity on the bronchial contractile response to the autacoid. These results suggest that silymarin has a protective effect in the early phase of allergic asthma, an effect, which may be related to a negative influence of the flavonoid on bronchial responsiveness to histamine. PMID- 11864646 TI - Glutamate mediated responses in isolated trachea preparations from control and ovalbumin sensitized guinea-pigs. AB - We investigated whether the glutamergic system plays a role in isolated trachea from control and ovalbumin-sensitized guinea-pigs. Electrical field stimulation induced contractile responses in control group, but electrical field stimulation produced relaxation responses in ovalbumin-challenged guinea-pigs. The responses induced by electrical field stimulation in both groups were completely abolished by tetrodotoxin, but unaffected by hexamethonium. DL-2-amino-5-phosphono-valeric acid (D-AP5) caused a concentration-dependent statistically significant inhibition in the contractile responses to electrical field stimulation50 (EFS50) in control guinea-pigs. But in the ovalbumin-challenged groups, D-AP5 did not cause any significant effect on the relaxation response to frequency of field stimulation (EFS50). N(G)-monmethyl-L-argine caused a significant inhibition in the relaxation effect of EFS50. L- and D-glutamate and N-methyl-D-aspartic acid (NMDA) alone had no effect on the resting tension on the trachea in both groups. Carbachol produced concentration-dependent contractile responses in ovalbumin challenged groups. These results suggested that responses to electrical field stimulation in control groups might be due to NMDA receptor-mediated release of any substance on prejunctional neurones and, alternatively, NMDA might exert a modulatory effect on any substance at prejunctional level. Also, responses to electrical field stimulation in ovalbumin-challenged guinea-pigs might not be mediated by NMDA but rather by increasing the production of nitric oxide by inducible nitric oxide synthase. PMID- 11864647 TI - Self-care self-efficacy, depression, and quality of life among patients receiving hemodialysis in Taiwan. AB - The purpose of this study is to examine relationships among self-care self efficacy, depression, and quality of life in 160 patients receiving hemodialysis. The study is a descriptive, correlational design. Measures include Strategies Used by People to Promote Health, the Geriatric Depression Scale, and the Quality of Life Index. Results indicate that self-care self-efficacy and depression are the significant predictors of quality of life after controlling for the effect of age. Self-care self-efficacy explains 47.5% of the variance (beta=0.52, p<0.001) and depression (beta=-0.29, p<0.001) explains an additional 5.5% of variance in quality of life. The study provides important information for health care providers as they design interventions for patients receiving hemodialysis. PMID- 11864648 TI - Violent death and the South African emergency nurse. AB - A violent death is the sudden and unexpected loss of life due to a violent means, and in South Africa, violent death due to interpersonal violence appears to be a huge problem with the majority of these clients being young adults in the prime of their life (Trauma Rev. 5(1) (1997) 1).The objective of the study was to explore the influence that the confrontation of violent death of their clients had on the emergency nurses' daily personal and professional lives. The participants described a number of physical and emotional ways in which their lives had been affected. PMID- 11864649 TI - The clinical learning environment and supervision by staff nurses: developing the instrument. AB - The aims of this study were (1) to describe students' perceptions of the clinical learning environment and clinical supervision and (2) to develop an evaluation scale by using the empirical results of this study. The data were collected using the Clinical Learning Environment and Supervision instrument (CLES). The instrument was based on the literature review of earlier studies. The derived instrument was tested empirically in a study involving nurse students (N=416) from four nursing colleges in Finland. The results demonstrated that the method of supervision, the number of separate supervision sessions and the psychological content of supervisory contact within a positive ward atmosphere are the most important variables in the students' clinical learning. The results also suggest that ward managers can create the conditions of a positive ward culture and a positive attitude towards students and their learning needs. The construct validity of the instrument was analysed by using exploratory factor analysis. The analysis indicated that the most important factor in the students' clinical learning is the supervisory relationship. The two most important factors constituting a 'good' clinical learning environment are the management style of the ward manager and the premises of nursing on the ward. The results of the factor analysis support the theoretical construction of the clinical learning environment modelled by earlier empirical studies. PMID- 11864650 TI - The home care experience as perceived by the caregivers of Chinese dialysis patients. AB - The caring issues perceived as crucial by caregivers in home-based dialysis, have never been systematically studied in any depth in Hong Kong. This research aimed to explore various characteristics of the home care experience perceived by caregivers. Thirty caregivers of home dialysis patients were interviewed using a phenomenological approach. Selection was by convenience. The interviews were conducted in the outpatient clinic following receipt of the informant's consent. The instrument consisted of two aspects designed to elicit the phenomena. The caregivers' responses seemed to indicate that caregiving is likely to have financial, emotional, social and health-related implications. PMID- 11864651 TI - Asthma management efficacy of school nurses in Taiwan. AB - As the key health care providers in school settings, the school nurses' asthma management efficacy is crucial to children's health and their continued participation in school learning activities. This article describes the psychometric testing of the asthma management efficacy scale (AMES) for use with school nurses. A cross-sectional survey design was used to assess the asthma management efficacy of 60 school nurses in Taipei, Taiwan. Factor analysis resulted in four factor domains including asthma medication administration, asthma pattern identification, school management behaviour, and monitoring peak expiratory flow rate which explained 76.4% of the variance in school nurses' asthma management efficacy. The school nurses' experience in performing school asthma management activities was positively correlated to their asthma management efficacy (r=0.33, p<0.05). School nurses who had experience with the inhaled asthma medicines had significantly higher efficacy scores on the medication administration subscale (t=-2.89, p<0.01) than did the school nurses who lacked this experience. School nurses who had experience in using peak expiratory flow meters had significantly higher efficacy scores on the total AMES (t=-1.90, p<0.05) and on the monitoring peak expiratory flow rate subscale (t=-5.37, p<0.001) than the school nurses who lacked this experience. Given the need to have nurses who are well prepared to provide asthma care in school settings, implications for nursing education, practice, and further research are discussed. PMID- 11864652 TI - Identification of child maltreatment while caring for them in a university hospital. AB - The purpose of the study was to look at how nurses and physicians of a university hospital rated their ability to identify child maltreatment while caring for those children. In this study, child maltreatment was defined as physical, psychological or sexual abuse or neglect of a child under the age of 18 by parents or caregivers. The total population of staff caring for children (N=513) in a university hospital were surveyed. Data were collected with a questionnaire developed for this study with reference to the literature. Altogether 317 questionnaires were returned, which yielded a response rate of 62%. The data were analysed using statistical methods and quantitative content analysis. Forty per cent of the respondents estimated that they had never cared for a maltreated child. Two-thirds of the respondents believed that they would be able to identify a child maltreatment case. The most distinct signs by which maltreatment could be identified were fractures, multiple bruises and the fact that the child had frequent injuries. The child's or parent's behaviour often aroused suspicion of maltreatment. Seventy-one per cent of the respondents rated the identification of maltreatment as rather difficult or difficult. Awkwardness of the phenomenon, the staff's pressure of work and relative unfamiliarity with the phenomenon were assessed as things that make the identification difficult. The fact that no jointly agreed guidelines were available for handling the matter was seen as a particular weakness. The respondents had fairly much theoretical knowledge about child maltreatment. However, child maltreatment is a multi-dimensional phenomenon that calls forth emotions. The development and improvement of practical nursing and medical care and of staff collaboration require that education be provided to different occupational groups and parties caring for children and that jointly agreed hospital-specific and regional models for operation be developed. PMID- 11864653 TI - An ergonomic approach to reducing back/shoulder stress in hospital nursing personnel: a five year follow up. AB - The purpose of the study was to determine the impact of an ergonomic program on perceived stress ratings, injury rates and patient care. After implementation of the ergonomic program, the perceived stress ratings by nursing staff were lower than those ratings at the control hospital and the patients felt more comfortable and secure during patient handling tasks than the patients at the control hospital. Eighteen months after ergonomic interventions, the back and shoulder injuries were reduced, and the lost workdays and restricted/transitional days were decreased. Five years after the implementation, the back and shoulder injuries continued to decrease as well as the lost workdays and restricted days. At the control hospital, the back and shoulder injury rates, the lost workdays, and the restricted days remained stable throughout the study period. PMID- 11864654 TI - Acute confusional episodes in elderly orthopaedic patients: the patients' actions and speech. AB - The aim of this study was to illuminate the pattern of confusional episodes with regard to the content of elderly patients' actions and speech. Fifty-one (11.2%) out of 457 people admitted to an orthopaedic clinic developed acute confusional state. Non-participant direct observations and daily clinical bedside follow-up observations were made during the confusional episodes, focusing on the patients' speech, actions and reactions with reference to situation, the surroundings, and the nurses' actions, including their verbal communication with the patient. The texts were analysed using manifest and latent content analysis and revealed disturbances in motor activity, speech and mood. 'Struggling to understand and to gain control when in a state of confusion and viewing oneself as being confused' was the main theme illuminated in the texts. In the struggle to achieve understanding and take control over themselves and what was currently taking place, the patients used a variety of strategies. They attempted to find clarity, to make a sensible story out of the present and to impute meaning to the present by using their life story. The findings indicated that the patients had a drive to achieve control when they were confused and that they were both aware and unaware of their state and tended to oscillate between being in a confused state and viewing their confusion from an outside perspective. The findings indicated that possibly the best approach is to support their inner drive to gain control. This means moving between confirmation of their emotional state and their perceptions and trying to introduce 'reality' when they realize their state of confusion. Thus the nurse--patient relationship perhaps needs to hold a balance between confirmation and introducing in reality. PMID- 11864655 TI - Nurses' perceptions of parental guidance in pediatric surgical pain relief. AB - The aim of this study was to describe nurses' perceptions of how they guide parents in the relief of their child's (aged 8-12 years) surgical pain in the hospital, and factors related to this function. The convenience sample consisted of 162 nurses working on the pediatric surgical wards of five university hospitals in Finland. The data was collected with a Likert-type instrument designed for this purpose. The results of this study indicated that nurses felt parents were mostly well informed about their child's surgical procedure, including both cognitive and sensory information, and about the non pharmacological methods employable for relieving their child's pain. However, some deficiency was identified in the preparatory information, as well as in the cognitive-behavioral and physical methods. Factors related to the nurses' background, such as age, education, work experience and the nurse's own experiences with prior hospitalizations of their children, appeared to have some effects on their perceptions regarding parental guidance. PMID- 11864656 TI - Adequate self-care of dialysed patients: a review of the literature. AB - The purpose of this article is to clarify the concept: 'adequate self-care of patients treated with hemodialysis (HD) or continuous ambulatory peritoneal dialysis (CAPD)'. This was done by the specification of Orem's general definition of self-care and a review of the literature. Adequate self-care behaviours for preventing and regulating pathological processes and related disabilities e.g. following dialysis and medication prescriptions as well as diet and fluid regimens are well documented. Adequate self-care behaviours aimed to promote personal well-being or development are less well documented. The authors suggest that use of the concept of adequate self-care would enlarge the scope of the disease management of HD and CAPD patients. Further research should focus on systematically investigating self-care activities of dialysed patients and their contribution to health and quality of life outcomes. PMID- 11864657 TI - Eating difficulties, assisted eating and nutritional status in elderly (> or = 65 years) patients in hospital rehabilitation. AB - This study describes frequencies and associations between eating difficulties, assisted eating and nutritional status in 520 elderly patients in hospital rehabilitation. Eating difficulties were observed during a meal and nutritional status was assessed with Subjective Global Assessment form. Eighty-two percent of patients had one or more eating difficulties, 36% had assisted eating and 46% malnutrition. Three components of eating were focused upon ingestion, deglutition, and energy (eating and intake). Deglutition and ingestion difficulties and low energy were associated with assisted eating, and low energy associated with malnutrition. Underestimation of low energy puts patients at risk of having or developing malnutrition. PMID- 11864658 TI - Moving into a care home: the role of adult children in the placement process. AB - Admission of an older person to a care home is widely recognised as a very stressful period for the family and one which, despite community care policy, is likely to be an increasingly common experience. Although there is a growing research base in this area, there have been few studies on the role of adult children in supporting their parents during this difficult transition. This paper reports on the third stage of a grounded theory study conducted in Sweden which explored the part played by adult children in the placement process. Data were collected from 13 adult children using in-depth semi-structured interviews and the results are compared with themes previously derived from interviews with 26 spouse carers. The analysis reveals important overlaps and differences and suggests the need for further research exploring the dynamics of the placement process. PMID- 11864660 TI - The risk takers. PMID- 11864661 TI - Behavior, hormones, and cardiovascular disease: interpretations and implications. PMID- 11864662 TI - Serum CA-125 in preoperative patients at high risk for endometriosis. AB - OBJECTIVE: To investigate the factors contributing to the elevated level of CA 125 in endometriosis and to study whether CA-125 assay is useful to identify women who require preoperative bowel preparation. METHODS: A total of 685 women undergoing surgery for endometriosis between July 1988 and June 1999 were studied. Preoperative serum CA-125 levels were compared between various pelvic conditions using F statistics. Multiple regression was employed to determine significant correlates of elevated serum CA-125, and the receiver operating characteristic curve was applied to assess the utility of serum CA-125 in preoperative preparation. Based on the two-sample Student t test, the sample size required to detect a difference in mean serum CA-125 levels of one-half of one standard deviation with a power of 90% when the sample size ratio of the two groups was 1:50 was 675 with a significance level of 5%. RESULTS: The mean serum CA-125 levels (IU/mL) for American Society of Reproductive Medicine stages I, II, III, and IV endometriosis were 18.8 +/- 0.9, 40.3 +/- 2.8, 77.1 +/- 3.5, and 182.4 +/- 14.0, respectively. CA-125 levels were significantly increased with advanced stages (P <.001, F test). Furthermore, serum CA-125 levels were significantly higher in patients with more extensive adhesions to the peritoneum, omentum, ovary, fallopian tube, colon, and cul-de-sac, or with ruptured endometrioma (P <.001, F test). We then classified patients with at least one of the three factors including dense omentum adhesion, ruptured endometrioma, and complete cul-de-sac obliteration as the high-risk group that required preoperative bowel preparation, and the others as the low-risk group. Receiver operating characteristic curve analyses set a cutoff point of 65 IU/mL, which gave a sensitivity of 76%, a specificity of 71%, a positive predictive value of 76%, and a negative predictive value of 93.2%. CONCLUSION: Our results suggest that preoperative CA-125 assay is useful to decide which women should receive preoperative bowel preparation. Endometriosis patients with preoperative CA-125 levels higher than 65 IU/mL are at high risk for severe pelvic adhesions that warrant thorough preoperative bowel preparation. PMID- 11864663 TI - Premenopausal social status and hormone exposure predict postmenopausal atherosclerosis in female monkeys. AB - OBJECTIVE: To determine whether premenopausal social subordination in female monkeys predicts postmenopausal atherosclerosis, and whether any such effect is altered by chronic exposure to contraceptive steroids or postmenopausal hormone replacement. METHODS: One hundred seventy-seven (177) premenopausal cynomolgus monkeys (Macaca fascicularis) housed in social groups of five or six were fed an atherogenic diet that, for half of the animals, also contained an oral contraceptive (OC). Individuals were judged socially dominant or subordinate based on behavioral observations. After 26 months animals were oophorectomized, biopsied for iliac atherosclerosis, and for the next 36 months were fed one of three atherogenic diets containing soy protein: 1) phytoestrogen-free; 2) phytoestrogens intact; and 3) phytoestrogen-free plus conjugated equine estrogens. Plasma lipids and menstrual cyclicity were also assessed. Finally, all animals were necropsied and the extent of atherosclerosis measured in the coronary and iliac arteries. RESULTS: The interaction of premenopausal social status and OC exposure predicted postmenopausal coronary artery atherosclerosis (P =.02). Subordinate animals not receiving OCs developed twice the coronary atherosclerosis of similarly untreated dominants (P <.01), an outcome mitigated by premenopausal OC exposure (P <.01). These effects occurred across postmenopausal treatment groups and independent of variation in plasma lipids. The same associations were observed in the iliac arteries, and, to a similar extent, both pre- and post-menopausally. Hormone data suggest that untreated premenopausal subordinates may have been estrogen deficient. CONCLUSION: Premenopausal social subordination exacerbates postmenopausal atherosclerosis, an effect possibly mediated by estrogen deficiency and shown here to be prevented by premenopausal OC exposure. These results occur irrespective of postmenopausal treatment. PMID- 11864664 TI - Benefits of soy isoflavone therapeutic regimen on menopausal symptoms. AB - OBJECTIVE: To examine the change in menopausal symptoms and cardiovascular risk factors in response to 4 months of daily 100-mg soy isoflavone in postmenopausal women. METHODS: In this double-blind, placebo-controlled study, 80 women were randomly assigned to isoflavone (n = 40) and placebo (n = 40) treatment. The menopausal Kupperman index was used to assess change in menopausal symptoms at baseline and after 4 months of treatment. Cardiovascular risk factors were assessed by evaluating plasma lipid levels, body mass index, blood pressure, and glucose levels in the participants. To examine the effects of this regime on endogenous hormone levels, follicle-stimulating hormone (FSH), luteinizing hormone (LH), and 17 beta-estradiol were measured. Transvaginal sonography was performed to quantify endometrial thickness. RESULTS: The data showed a decrease in menopausal symptoms (P <.01, paired t test, two-tailed, between baseline and isoflavone groups, and P <.01, unpaired t test, between placebo and isoflavone groups). Total cholesterol and low-density lipoprotein decreased significantly in the isoflavone group compared with the baseline or placebo group (P <.001, paired t test, two-tailed, between baseline and isoflavone groups, and P <.01, unpaired t test, between placebo and isoflavone groups). The isoflavone treatment appeared to have no effect on blood pressure, plasma glucose, and high-density lipoprotein and triglyceride levels. CONCLUSION: This study suggests that isoflavone 100-mg regime treatment may be a safe and effective alternative therapy for menopausal symptoms and may offer a benefit to the cardiovascular system. PMID- 11864665 TI - Has the use of routine episiotomy decreased? Examination of episiotomy rates from 1983 to 2000. AB - OBJECTIVE: To determine if practice patterns have been altered by the large body of literature strongly advocating the selective use of episiotomy. METHODS: An electronic audit of the medical procedures database at Thomas Jefferson University Hospital from 1983 to 2000 was completed. Univariate and multivariable models were computed using logistic regression models. RESULTS: Overall episiotomy rates in 34,048 vaginal births showed a significant reduction from 69.6% in 1983 to 19.4% in 2000. Significantly decreased risk of episiotomy was seen based upon year of childbirth (odds ratio [OR] 0.87, 95% confidence interval [CI] 0.86, 0.87), black race (OR 0.29, 95% CI 0.28, 0.31), and spontaneous vaginal delivery (OR 0.40, 95% CI 0.36, 0.45). Increased association with episiotomy was seen in forceps deliveries (OR 4.04, 95% CI 3.46, 4.72), and with third- or fourth-degree lacerations (OR 4.87, 95% CI 4.38, 5.41). In deliveries with known insurance status, having Medicaid insurance was also associated with a decreased episiotomy risk (OR 0.59, 95% CI 0.54, 0.64). CONCLUSION: There was a statistically significant reduction in the overall episiotomy rate between 1983 and 2000. White women consistently underwent episiotomy more frequently than black women even when controlling for age, parity, insurance status, and operative vaginal delivery. PMID- 11864666 TI - Variations in antenatal corticosteroid therapy: a persistent problem despite 30 years of evidence. AB - OBJECTIVE: To document current use of antenatal corticosteroid therapy in a large cohort of Canadian preterm infants admitted to neonatal intensive care units, and to assess the impact of variations in use on neonatal outcomes. METHODS: The study subjects included 11,440 infants less than 38 weeks' gestation who were admitted to 17 Canadian Neonatal Network intensive care units from January 1996 to October 1997. Data analyses were conducted separately for infants less than 24 weeks' gestation, 24-34 weeks' gestation, and over 34 weeks' gestation. Logistic regression analysis was used to model the examined relationships, controlling for patient characteristics. RESULTS: The incidence of antenatal corticosteroid treatment was 42% for infants less than 24 weeks' gestation, 59% for infants 24 34 weeks' gestation, and 10% for infants over 34 weeks' gestation. Antenatal corticosteroid treatment was associated with reduced risk for neonatal mortality and respiratory distress syndrome, but not for infants over 34 weeks' gestation. Significant institutional variations in antenatal corticosteroid use were present among both inborn and outborn infants. Increased antenatal corticosteroid treatment for infants 24-34 weeks' gestation can potentially reduce the number of neonatal deaths by 41 cases (10%) and respiratory distress syndrome by 90 cases (3%) among participating hospitals. CONCLUSION: Wide institutional differences persist in the incidence of antenatal corticosteroid treatment for women expected to give birth preterm. Increased use of antenatal corticosteroids for preterm deliveries can reduce neonatal mortality in Canada by up to 10%. PMID- 11864667 TI - Risk factors for difficult delivery in nulliparas with epidural analgesia in second stage of labor. AB - OBJECTIVE: To identify risk factors for difficult delivery among nulliparas in the second stage of labor with continuous epidural analgesia, and to develop a multivariable model that is predictive of difficult delivery. METHODS: The database is derived from a multicenter randomized trial of delayed pushing for nulliparous women under continuous infusion epidural. Members of this cohort (n = 1862) were randomly divided into two groups: a "Model Development" and a "Model Validation" group. We used univariate and multivariable techniques to assess associations between anthropometric, sociodemographic, and obstetric variables and difficult delivery. RESULTS: With the referent defined as the category of lesser risk, the developed model showed that the risk of difficult delivery was increased for women with height less than 160 cm (odds ratio [OR] 2.1, 90% confidence interval [CI] 1.2, 3.4), prepregnancy weight greater than 65 kg (OR 1.6, 90% CI 1.0, 2.6), age greater than or equal to 35 years (OR 3.0, 90% CI 1.1, 8.1), and gestational age greater than or equal to 41 weeks (OR 1.8, 90% CI 1.1, 2.8). Induction of epidural analgesia late in labor (greater than or equal to 6 cm) was associated with a higher risk of difficult delivery than induction between 3 and 5 cm (OR 1.9, 90% CI 1.3, 2.8). An interval of greater than or equal to 360 minutes between epidural induction and full dilatation increased the risk of difficult delivery (OR 3.8, 90% CI 1.5, 9.5). Fetal station above +2 at full dilatation and a posterior fetal position were both strongly associated with difficult delivery (OR 2.7, 90% CI 1.4, 5.0, and OR 11.2, 90% CI 4.9, 25.6, respectively). For the multivariable predictive model, when the sensitivity was 57%, the specificity was 75%, and the positive predictive value was 35%. CONCLUSION: Our observations concerning maternal characteristics and obstetric variables are consistent with previous observations with the exception of time of induction of the epidural. The predictive model may be useful in defining high risk populations for subsequent intervention studies designed to assess approaches to reduce difficult delivery. PMID- 11864668 TI - Over-the-counter antifungal drug misuse associated with patient-diagnosed vulvovaginal candidiasis. AB - OBJECTIVE: To estimate what proportion of symptomatic women purchasing over-the counter antifungal products for immediate treatment of presumed vulvovaginal candidiasis have vulvovaginal candidiasis or other genitourinary conditions. METHODS: A time-location sample of 95 symptomatic women who purchased and presented with an over-the-counter antifungal product for immediate and personal treatment of presumed vulvovaginal candidiasis were evaluated by clinical examination and pertinent laboratory tests. The percentage of women diagnosed having vulvovaginal candidiasis or other conditions, proportions of women with vulvovaginal candidiasis compared between groups with and without a prior diagnosis of vulvovaginal candidiasis, and groups that read or did not read the over-the-counter package label were assessed. RESULTS: The actual diagnoses for women who self-diagnosed vulvovaginal candidiasis were: vulvovaginal candidiasis 33.7%, bacterial vaginosis 18.9%, mixed vaginitis 21.1%, normal 13.7%, other diagnoses 10.5%, and trichomonas vaginitis 2.1%. Women with a previous clinically based diagnosis of vulvovaginal candidiasis were not more accurate in diagnosing vulvovaginal candidiasis than women without a prior clinical diagnosis (chi(2) = 0.27, P =.6). Women who read the package label were no more likely to have vulvovaginal candidiasis than were women who did not read the label (Fisher exact test, P =.39). CONCLUSION: Many women who self-diagnose and use an over-the counter product for treatment of presumed vulvovaginal candidiasis do not have vulvovaginal candidiasis. A history of a previous clinically based diagnosis of vulvovaginal candidiasis and reading the package label do not help women self diagnose vulvovaginal candidiasis properly. Ready access to these products is associated with wasted financial expenditures, unfulfilled expectations, and a delay in correct diagnosis for a substantial number of women. PMID- 11864669 TI - Predictors of subsequent surgery for uterine leiomyomata after abdominal myomectomy. AB - OBJECTIVE: To study factors associated with an increased risk of subsequent surgery in a cohort of women undergoing abdominal myomectomy. METHODS: We followed 65 women undergoing abdominal myomectomy performed by the same experienced surgeon for a mean of 83.6 +/- 35.0 months to assess the occurrence of both laparotomies and minimally invasive surgeries for uterine leiomyomas. Surgical and pathologic variables from the initial myomectomy as well as information on sociodemographic and anthropometric variables collected during a subsequent survey were correlated with the need for further surgery. RESULTS: Women with uterine size greater than 12 menstrual weeks had a substantially reduced risk of undergoing a second surgery (multivariate hazard ratio 0.1, 95% confidence interval 0.01, 0.4) compared with women having smaller uteri. Weight gain in excess of 30 pounds since age 18, relative to weight gain of 10 or fewer pounds is also associated with an increased risk of recurrent surgery (multivariate hazard ratio 4.8, 95% confidence interval 1.2, 18.5). CONCLUSION: Women with uterine size less than 12 menstrual weeks at the time of abdominal myomectomy may be at increased risk of second surgery. Weight gain after age 18 may also modify the risk of recurrent surgery. These changes may be related to the pathogenic mechanisms underlying myoma formation and growth. PMID- 11864670 TI - Magnetic resonance imaging of normal levator ani anatomy and function. AB - OBJECTIVE: To evaluate the anatomy and function of the levator ani in normal women by dynamic magnetic resonance imaging. METHODS: Twelve asymptomatic, nulliparous, premenopausal women with no previous pelvic surgery underwent a dynamic magnetic resonance imaging scan of their pelvis. The origin, orientation, thickness, and function of the two components of the levator ani were studied. RESULTS: The ileococcygeus is a thin muscle with an upward convexity. It slopes forward and medially. It is of variable thickness (mean thickness 2.9 mm, standard deviation 0.8 mm). There are apparent gaps in the muscle diaphragm and at its site of origin from the obturator fascia. The puborectalis is a thicker muscle. It is shaped like a belt encasing the pelvic organs. It is taller posteriorly than anteriorly. It is not attached to the bladder neck, but the midurethra and lower urethra lie in close proximity to it. The puborectalis moves dorsoventrally, whereas the ileococcygeus moves craniocaudally. CONCLUSION: The levator ani is not a single muscle but has two functional components that vary in thickness, origin, and function. The ileococcygeus has a mainly supportive function, whereas the puborectalis has a sphincteric function. Gaps in the diaphragmatic portion of the ileococcygeus are a normal finding. Individual components of the levator ani may be prone to different types of childbirth trauma and should therefore be assessed separately when planning rehabilitation. PMID- 11864671 TI - Oral misoprostol before office endometrial biopsy. AB - OBJECTIVE: To evaluate oral misoprostol use before office endometrial biopsy. METHODS: Forty-two nonpregnant women aged 35-77 years were randomized to a prospective, double-blind study to receive either 400 microg oral misoprostol or placebo 3 hours before office endometrial biopsy. Misoprostol effects were assessed by 1) cervical resistance, 2) ease of performing the endometrial biopsy, 3) success rate of obtaining an endometrial biopsy, 4) pain intensity associated with the endometrial biopsy, and 5) adverse clinical side effects. RESULTS: Patients in the misoprostol group experienced significantly (P <.01) more pain associated with the endometrial biopsy. The observed power to detect this difference in misoprostol-placebo comparison using the Wilcoxon rank sum test at 0.05 level of significance is 89%. In addition, significantly (P <.05) more patients had the adverse side effect of uterine cramping at 1.5 hours after medication ingestion in the misoprostol group. The observed power to detect this difference is 98%. There were no differences between the misoprostol and placebo groups in cervical resistance, ease of performing the biopsy, success rate for obtaining an endometrial biopsy, or adverse side effects at 3 hours post medication ingestion. CONCLUSION: Oral misoprostol 400 microg caused more uterine cramping and pain in nonpregnant women undergoing office endometrial biopsy when given 3 hours before biopsy attempt. No other cervical effects were noted. PMID- 11864672 TI - Preeclampsia in multiple gestation: the role of assisted reproductive technologies. AB - OBJECTIVE: To estimate the relationship of assisted reproductive technologies and ovulation-inducing drugs with preeclampsia in multiple gestations. METHODS: This historical cohort study was conducted on 528 multiple gestations from a Colorado health maintenance organization. Using univariate and logistic regression analysis, we determined if women who conceived a multiple gestation as a result of assisted conception were at a greater risk of preeclampsia than those who conceived spontaneously. RESULTS: Between January 1994 and November 2000, there were 330 unassisted and 198 assisted multiple gestations. Sixty-nine multiple gestations followed assisted reproductive technologies (in vitro fertilization and gamete intrafallopian transfer). Human menopausal gonadotropins and clomiphene citrate were associated with 38 and 91 of the multiple gestations, respectively. Compared with unassisted multiple gestations, the relative risk of mild or severe preeclampsia among mothers who received assisted reproductive technologies was 2.7 (95% confidence interval [CI] 1.7, 4.7) and 4.8 (CI 1.9, 11.6), respectively. Adjusted for maternal age and parity, women who received assisted reproductive technologies were two times more likely to develop preeclampsia (odds ratio 2.1, CI 1.1, 4.1) compared with those who conceived spontaneously. The adjusted odds ratios of nulliparity and maternal age for preeclampsia were 2.1 (CI 1.3, 3.4) and 1.1 (CI 1, 1.1), respectively. Although the incidence of preeclampsia was greater in mothers who received clomiphene citrate and human menopausal gonadotropins, this association did not reach statistical significance at the P <.05 level. CONCLUSION: Women who conceive multiple gestations through assisted reproductive technologies have a 2.1-fold higher risk of preeclampsia than those who conceive spontaneously. PMID- 11864673 TI - A randomized trial of intrapartum analgesia in women with severe preeclampsia. AB - OBJECTIVE: To estimate whether the cesarean delivery rate differs between women with severe preeclampsia who receive intrapartum epidural analgesia versus patient-controlled intravenous opioid analgesia. METHODS: Women with severe preeclampsia at at least 24 weeks' gestation were randomly assigned to receive either intrapartum epidural (n = 56) versus patient-controlled intravenous opioid analgesia (n = 60), and each was administered by a standardized protocol. The sample size was selected to have 80% power to detect at least a 50% difference in the predicted intergroup cesarean delivery rates. Data were analyzed by intent to treat. RESULTS: Selected maternal characteristics and neonatal outcomes were similar in the two groups. The cesarean delivery rates in the epidural group (18%) and the patient-controlled analgesia group (12%) were similar (P =.35). Women who received epidural analgesia were more likely to require ephedrine for the treatment of hypotension (9% versus 0%, P =.02), but their infants were less likely to require naloxone at delivery (9% versus 54%, P <.001). Epidural analgesia provided significantly better pain relief as determined by a visual analogue intrapartum pain score (P <.001) and a postpartum pain management survey (P =.002). CONCLUSION: Compared with patient-controlled intravenous opioid analgesia, intrapartum epidural analgesia did not significantly increase the cesarean delivery rate in women with severe preeclampsia at our level III center, and it provided superior pain relief. PMID- 11864674 TI - Indicators of cocaine exposure and preterm birth. AB - OBJECTIVE: To identify predictors of cocaine exposure during pregnancy, using hair and urine assays and self-report, and the association with preterm birth. METHODS: A nested case-control study was conducted in a cohort of 2611 black and white women enrolled in prenatal care in central North Carolina. Cocaine exposure was ascertained by self-report (263 cases, 612 controls), urine assays at 24-29 weeks' gestation (226 cases, 564 controls) and postpartum (160 cases, 408 controls), and postpartum hair assays (169 cases, 435 controls). The major metabolite of cocaine, benzoylecgonine, was measured in urine. Cocaine and benzoylecgonine were measured in hair. RESULTS: Cocaine exposure was identified in 2% based on self-report, 5-6% based on urine assays, and 13-15% based on hair assays. Black ethnicity, lower education, and poverty were strongly predictive of positive hair assays. Hair cocaine and benzoylecgonine were not associated with preterm birth, with the possible exception of higher levels of cocaine and benzoylecgonine and birth before 34 weeks' completed gestation. The urine screen at 24-29 weeks' gestation also gave some indication of a possible association (odds ratio 1.7, 95% confidence interval 0.9, 3.5). CONCLUSION: The study corroborates the incompleteness of cocaine exposure assessment by self-report and urine screens relative to hair assays. The strong demographic predictors of exposure suggest where intervention efforts should be targeted. The most sensitive markers of exposure, hair cocaine and benzoylecgonine, are not associated with preterm birth, perhaps because they reflect different patterns of cocaine exposure than the other measures. PMID- 11864675 TI - Antepartum, intrapartum, and neonatal significance of exercise on healthy low risk pregnant working women. AB - OBJECTIVE: To evaluate the influence of exercise on maternal and perinatal outcome in a low-risk healthy obstetric population. METHODS: We conducted a prospective observational study of low-risk healthy women exercising during their pregnancy. An extensive questionnaire collected antepartum, intrapartum, and postpartum patient information on 750 women. The women were divided into four groups based on exercise level during pregnancy. RESULTS: There were no differences among groups for maternal demographic characteristics, antenatal illnesses, stress, social support, or smoking. Heavily exercising women were older (P =.042), had higher incomes (P =.001), and were exercising more at conception (P =.001). Women who did more exercise were more likely to need an induction of labor (P =.033, relative risk 1.84, 95% confidence interval 1.05, 3.20), induction or augmentation with oxytocin (P =.015, relative risk 1.53, 95% confidence interval 1.19, 1.97), and had longer first-stage labors (P =.032) resulting in longer total labors (P =.011). The difference in the length of first stage labor was even greater if the no-exercise group was compared with the strongly exercising group (P =.009, relative risk 1.38, 95% confidence interval 0.16, 2.60). Fewer umbilical cord abnormalities (P =.034) were observed with exercise, but exercising women had more colds and flu (P =.008). Heavily exercising women had smaller infants (mean difference 86.5 g) compared with sedentary women. CONCLUSION: Exercise in working women is associated with smaller babies, increased number of inductions and augmentations of labor, and longer labors. Colds and flu are more frequent in exercising women. PMID- 11864676 TI - Is urethral mobility really being assessed by the pelvic organ prolapse quantification (POP-Q) system? AB - OBJECTIVE: To estimate the relationship between Q-tip measurement of urethral hypermobility and visual assessment of the urethrovesical junction as assessed by points Aa and Ba of the pelvic organ prolapse quantification (POP-Q) system. METHODS: A total of 274 patients with pelvic organ prolapse or urinary incontinence had preoperative Q-tip test straining angles and POP-Q staging measurements. By the Q-tip test, urethral hypermobility was defined as a straining angle of 30 degrees or greater relative to the horizontal. As defined in the POP-Q system, point Aa is located in the midline of the anterior vaginal wall 3 cm from the external urethral meatus and represents the urethrovesical junction. Point Ba represents the most dependent position of the anterior vaginal wall. The correlation between point Aa of the POP-Q system and the Q-tip test was assessed using the Spearman correlation coefficient. Similar assessments were made for point Ba. RESULTS: Mean age of the 274 subjects was 58.5 +/- 11.8 years; mean parity was 3.1 +/- 1.6. A total of 104 patients reported prior surgery for prolapse or incontinence. Mean Q-tip straining angle was 61 +/- 20 degrees; 258 (94%) had urethral hypermobility. Values of point Aa ranged from -3 cm to +3 cm, with median 0 cm. The correlation coefficient between the Q-tip straining angle and point Aa was r = 0.47 (P <.001). Urethral hypermobility was observed in 95% of patients with stage II prolapse at point Aa and in 100% of patients with stages III and IV prolapse at point Aa. The correlation coefficient between the Q tip straining angle and point Ba was r = 0.32 (P <.001). CONCLUSION: Although the correlation between the Q-tip straining angle and point Aa of the POP-Q was moderately strong, one value cannot be predicted from the other. The Q-tip test may be unnecessary in patients with stages II, III, and IV prolapse at point Aa as virtually all such patients demonstrate urethral hypermobility. PMID- 11864677 TI - Placental cord insertion and birth weight discordancy in twin gestations. AB - OBJECTIVE: To evaluate whether abnormal umbilical cord insertion (UCI) into the placenta is a risk factor for birth weight discordancy in twin gestations. METHODS: Pathology records of all liveborn twins delivered between January 1993 and June 1996 were reviewed. The information collected included gestational age at delivery, birth weight, gross placental pathology, and placental UCI velamentous, marginal, or disc. Discordancy in birth weight was defined as an intrapair difference of at least 20%. Analyses were stratified on placental chorionicity. Odds ratios and 95% confidence intervals for birth weight discordancy were calculated based on the presence of an abnormal (velamentous or marginal) placental UCI relative to normal (disc) UCI on both placentae, after adjusting for potential confounders. RESULTS: There were 447 twin pairs identified. Dichorionic diamniotic placentation was present in 358 pairs (80.1%), monochorionic diamniotic in 84 (18.8%), and monochorionic monoamniotic in five (1.1%). There was a 13-fold increase in the risk of birth weight discordancy in monochorionic diamniotic twins in the presence of a velamentous UCI (odds ratio 13.5, 95% confidence interval 1.4, 138.4), with a rate of birth weight discordancy of 46%. This relationship was not demonstrated in dichorionic diamniotic twins (odds ratio 1.0, 95% confidence interval 0.3, 3.5). CONCLUSION: Birth weight discordancy in twins is a different entity depending on chorionicity. The substantial increase in birth weight discordancy in monochorionic diamniotic twins that accompanies velamentous UCI underscores the need for prenatal detection and increased surveillance in these twins. PMID- 11864678 TI - Prenatal care and black-white fetal death disparity in the United States: heterogeneity by high-risk conditions. AB - OBJECTIVE: To determine the impact of prenatal care in the United States on the fetal death rate in the presence and absence of obstetric and medical high-risk conditions, and to explore the role of these high risk conditions in contributing to the black-white disparity. METHODS: This is a population-based, retrospective cohort study using the national perinatal mortality data for 1995-1997 assembled by the National Center for Health Statistics. Fetal death rate (per 1000 births) and adjusted relative risks were derived from multivariable logistic regression models. RESULTS: Of 10,560,077 singleton births, 29,469 (2.8 per 1000) resulted in fetal death. Fetal death rates were higher for blacks than whites in the presence (4.2 versus 2.4 per 1000) and absence (17.2 versus 2.5 per 1000) of prenatal care. Lack of prenatal care increased the (adjusted) relative risk for fetal death 2.9-fold in blacks and 3.4-fold in whites. Blacks were 3.3 times more likely to have no prenatal care compared with whites. Over 20% of all fetal deaths were associated with growth restriction and placental abruption, both in the presence and absence of prenatal care. Lack of prenatal care was associated with increased fetal death rates for both blacks and whites in the presence and absence of high-risk conditions. CONCLUSION: In the Unites States, strategies to increase prenatal care participation, especially among blacks, are expected to decrease fetal death rates. PMID- 11864679 TI - Intrauterine growth restriction. AB - Fetal intrauterine growth restriction presents a complex management problem for the clinician. The failure of a fetus to achieve its growth potential imparts a significantly increased risk of perinatal morbidity and mortality. Consequently, the obstetrician must recognize and accurately diagnose inadequate fetal growth and attempt to determine its cause. Growth aberrations, which are the result of intrinsic fetal factors such as aneuploidy and multifactorial congenital malformations, and fetal infection, carry a guarded prognosis. However, when intrauterine growth restriction is caused by placental abnormalities or maternal disease, the growth aberration is usually the consequence of inadequate substrates for fetal metabolism and, to a greater or lesser degree, decreased oxygen availability. Careful monitoring of fetal growth and well-being, combined with appropriate timing and mode of delivery, can best ensure a favorable outcome. Ultrasound evaluation of fetal growth, behavior, and measurement of impedance to blood flow in fetal arterial and venous vessels form the cornerstone of evaluation of fetal condition and decision making. PMID- 11864680 TI - Vacuum-assisted closure in the treatment of complex gynecologic wound failures. AB - BACKGROUND: Complex wound failures are a source of significant morbidity and mortality. They are costly and time consuming to treat, and may evolve into chronic, debilitating conditions. Vacuum-assisted closure is a novel wound healing technique applying subatmospheric pressure to wounds to expedite healing. CASES: We report the successful use of vacuum-assisted closure therapy on three patients on a gynecologic oncology service with complex wound failures of various chronicity. In all cases, vacuum-assisted closure therapy was well tolerated and demonstrated efficacy within 48 hours of initiation. CONCLUSION: We conclude that vacuum-assisted closure therapy should be included in the armamentarium of the gynecologist addressing complex wound failures. PMID- 11864681 TI - Uterine compression sutures: surgical management of postpartum hemorrhage. AB - BACKGROUND: It has been estimated that worldwide, over 125,000 women die of postpartum hemorrhage each year. The traditional management of this condition includes the use of oxytocics, such as oxytocin, ergometrine, and prostaglandins, before proceeding to ligation of the internal iliac arteries and even hysterectomy. The B-Lynch technique is a surgical procedure that may be used to arrest postpartum hemorrhage resulting from uterine atony. CASES: This paper describes simple modifications of this technique that make this procedure less complex to perform. Three clinical case scenarios illustrate the context in which the sutures may be used. CONCLUSION: Compression sutures placed into the postpartum uterus may provide a simple first surgical step to control bleeding when routine oxytocic measures have failed. We suggest that the technique we have described is a simple procedure and should be tried before more complex interventions are used. PMID- 11864682 TI - Pyometria after hemostatic square suture technique. AB - BACKGROUND: The square suture technique to achieve hemostasis for postpartum hemorrhage has been described in the medical literature. In a prior report of 23 cases, there were no complications from the technique. CASE: A young woman with an gravida 1 para 0 with an uncomplicated prenatal course presented at 40 weeks' estimated gestational age in active labor. Cesarean delivery was performed for arrest of dilatation and was complicated by postpartum hemorrhage unresponsive to medical therapy and uterine artery ligation. Several hemostatic square sutures were placed, obtaining hemostasis. The patient presented 4 weeks postpartum with pain and mucopurulent discharge. Pyometrium was discovered at the time of dilation and curettage. CONCLUSION: Pyometrium may result from the use of hemostatic square suture technique for control of postpartum hemorrhage. PMID- 11864683 TI - Hysteroscopy in the evaluation and treatment of mucinous adenocarcinoma. AB - BACKGROUND: Hysteroscopy to evaluate abnormal uterine bleeding is gaining popularity. The standard methods of evaluation, endocervical curettage, and endometrial biopsy frequently diagnose adenocarcinoma without determining location. Because the treatments of endometrial and endocervical cancers are different, knowing the neoplastic origin is desirable. CASES: Two postmenopausal women were referred for abnormal uterine bleeding. Endometrial biopsies were consistent with mucinous adenocarcinoma without distinction between cervical and endometrial sites. Endocervical curettages were inconclusive. Both patients underwent hysteroscopy at the time of exploratory laparotomy, which revealed the location of the adenocarcinomas, one in the endocervix and one in the uterine fundus. CONCLUSION: The location of adenocarcinoma may be further clarified by the use of intraoperative hysteroscopy, which can aid in determining surgical treatment. PMID- 11864684 TI - Pneumoamnion and pregnancy loss after second-trimester laparoscopic surgery. AB - BACKGROUND: Midtrimester laparoscopic surgery is considered safe; however, both maternal and fetal complications can occur. We report a case of midtrimester pregnancy loss with pneumoamnion after laparoscopic appendectomy. CASE: A young woman presented at 21 weeks' gestation with a 3-day history of nausea and severe abdominal pain. Diagnostic laparoscopy was performed with normal appendix and pelvic anatomy noted. Worsening pain prompted a repeat laparoscopic evaluation. A small defect in the uterine serosa was noted, which presumably resulted from inadvertent Veress needle injury. Abdominal computerized tomography was performed, which demonstrated a pneumoamnion. Spontaneous rupture of membranes and labor ensued, resulting in the delivery of a stillborn fetus. CONCLUSION: Inadvertent introduction of the Veress needle into the gravid uterus with subsequent pneumoamnion represents a catastrophic complication of midtrimester laparoscopic surgery. PMID- 11864685 TI - Fetal pulse oximetry. PMID- 11864687 TI - Falsely elevated human chorionic gonadotropin leading to unnecessary therapy. PMID- 11864690 TI - Cervical cancer screening by simple visual inspection after acetic acid. PMID- 11864695 TI - Should we allow a trial of labor after a previous cesarean for dystocia in the second stage of labor? PMID- 11864692 TI - Tocolysis with nifedipine or beta-adrenergic agonists: a meta-analysis. PMID- 11864697 TI - Fetal abdominal wall defects and mode of delivery: a systematic review. PMID- 11864698 TI - ACOG Committee opinion #270. Bone density screening for osteoporosis. American College of Obstetricians and Gynecologists. PMID- 11864700 TI - Fibrin precursors, intermediaries for hemostasis in the clot war. PMID- 11864701 TI - Effect of delivery modalities on the physiologic inhibition system of coagulation of the neonate. AB - The perinatal period is associated with an increased incidence of thromboembolic complications, which may occur in both the maternal and fetal circulation in otherwise normal and healthy adults and fetuses, and this may be related to the activation of the coagulation system at the time of parturition. The risk of these complications is generally much higher in neonates, who have decreased activity of the physiologic inhibition system of coagulation (PISC), including protein C, protein S and antithrombin, in comparison with adults. Therefore, any additional obstetric iatrogenic factors could predispose the neonate to an increased risk of thromboembolic complications. The aim of this study was to evaluate the influence of modality of delivery (spontaneous vaginal delivery vs. elective caesarian section) on the neonatal PISC factor (protein C, protein S and antithrombin) levels and the fibrinolytic system (plasminogen and fibrinogen levels). We studied 41 consecutive healthy newborns, 18 delivered vaginally (mean gestational age 39.7 +/- 0.8) and 23 by elective caesarian section (mean gestational age 38.5 +/- 0.7). Plasma samples were collected from the umbilical cord at birth. AT activity, protein C antigen and activity, total and free protein S antigen, fibrinogen concentration and plasminogen activity were tested. Among PISC factors studied in cord blood of infants born after vaginal delivery, protein C antigen levels and antithrombin activity were statistically higher (41.3 +/- 9.4 vs. 33.9 +/- 7.2 and 58.5 +/- 10.0 vs. 48.4 +/- 12.7, respectively; P<.01), while free protein S was significantly lower (36.8 +/- 11.6 vs. 46.4 +/- 12.5; P<.05) than in newborns delivered by caesarian section. Cord blood plasminogen and fibrinogen were elevated in vaginally delivered neonates in comparison to those delivered by caesarian section, but the difference was not statistically significant. Our data show that the labor stress of vaginal delivery may play a role in influencing the levels of some PISC factors in the cord blood of full-term neonates. In newborns with coagulation disorders, separate reference ranges in coagulation screening tests should be possibly needed depending on the delivery modality. PMID- 11864702 TI - The elevated plasma lipoprotein(a) concentrations in preeclampsia do not precede the development of the disorder. AB - We sought to determine whether maternal plasma lipoprotein(a) [Lp(a)] levels are elevated in the second trimester, before the development of preeclampsia and other obstetrical complications, in women at risk. In the first part of the study (cross-sectional), plasma concentrations of Lp(a) were compared among 16 women with preeclampsia, 35 normotensive pregnant women and 18 healthy nonpregnant women. In the second part (nested case-control), blood samples were collected prospectively from 82 women at risk of preeclampsia, at 14-24 weeks of gestation, and Lp(a) levels were compared between those in whom preeclampsia or other obstetrical complications developed and those in whom they did not. In the cross sectional study, plasma concentrations of Lp(a) were significantly higher in women with preeclampsia than in normotensive pregnant and healthy nonpregnant women (41 +/- 31 vs. 24 +/- 16 and 15 +/- 10 mg/dl, respectively; P=.001). Of the 82 women in the second part of the study, 9 (11%) developed preeclampsia and 19 (23%) had complications such as intrauterine growth restriction, preterm delivery and fetal or neonatal loss. There were no differences in plasma Lp(a) concentrations between the women with preeclampsia and those without complications, though Lp(a) levels were significantly higher in women with other complications than in those with either preeclampsia or uncomplicated pregnancies (40 +/- 29 vs. 17 +/- 13 or 28 +/- 18 mg/dl, respectively; P=.05). In conclusion, elevated plasma levels of Lp(a), associated with clinically established preeclampsia, are not detected before the appearance of the disorder in pregnant women at risk. PMID- 11864703 TI - Increased levels of endothelial haemostatic markers in patients with coronary heart disease. AB - von Willebrand factor (vWF), thrombomodulin and tissue plasminogen activator antigen (tPAag) are regarded as markers of endothelial activation and/or damage. Elevated levels have been associated with atherosclerotic disease states. The aim of the present study was to compare the levels of vWF, thrombomodulin and tPAag in patients with coronary heart disease (CHD) and matched healthy individuals to see if they discriminated significantly between the study groups also after adjustment for established CHD risk factors. Patients (n=193) in various stages of CHD and matched controls (n=193) were included. To evaluate possible influence of acute phase reaction, reinvestigation was performed after 6 months. We observed elevated levels of vWF (P<.001) and tPAag (P<.001) but not thrombomodulin (P=.082) in CHD patients when compared to controls, still statistically significant after 6 months and also after adjustment for established risk factors. Our results indicate that vWF and tPAag but not thrombomodulin in the present population are independent markers of atherosclerosis. PMID- 11864704 TI - Elevated plasma levels of the factor Xa-TFPI complex in cancer patients. AB - We have previously shown that cancer patients with solid tumour disease have increased plasma levels of both the free and total forms of the coagulation inhibitor, tissue factor pathway inhibitor (TFPI), whereas patients with leukemia and related blood malignancies have levels within the normal range. We now report that also the median plasma levels of the Factor Xa (FXa)-TFPI complex were significantly higher in patients with solid tumours, compared to patients with haematological malignancy and healthy controls. There were significant positive correlations between the FXa-TFPI complex and total TFPI antigen (r=.47, P=.001) and TFPI activity (r=.33, P<.023). In plasma samples from patients with solid tumours, the ratio between the FXa-TFPI complex and free TFPI was 3.4 times higher than in patients with haematological malignancies. Increased levels of the FXa-TFPI complex in solid tumour disease may reflect both increased FXa generation and the increased TFPI concentration in the patients. It is speculated that high levels of the inhibitory FXa-TFPI complex in cancer patients may protect against microthrombosis and organ failure, which are relatively rare in cancer despite long-lasting hypercoagulation. PMID- 11864705 TI - Increased plasma fibronectin levels in patients with acute myocardial infarction complicated with left ventricular thrombus. AB - Fibronectin is a polymorphic and multifunctional glycoprotein that plays a wide ranging role in hemostasis. In this study, it was aimed to determine plasma fibronectin levels and evaluate its possible role in left ventricular (LV) thrombus formation following acute myocardial infarction (AMI). We have determined clinical, echocardiographic, and biochemical parameters in 97 consecutive patients (aged 59 +/- 13; 87 men/10 women) with first anterior AMI. Two-dimensional echocardiography was performed on Days 1, 3, 7, 15, and 30. Blood samples were obtained within 24-48 h after the onset of symptoms. The study also included 30 healthy control subjects. Plasma fibronectin levels were significantly higher in patients with AMI than control subjects (38 +/- 13 vs. 25.2 +/- 8.7 mg/dl, P=.0001). LV thrombus was detected in 20 (20.6%) of 97 patients. Plasma fibronectin levels were significantly higher in patients with LV thrombus (Group 2) than in patients without LV thrombus (Group 1) (44.5 +/- 11 vs. 36.1 +/-13.4 mg/dl, P=.01). Although univariate analysis showed that plasma fibronectin levels were higher in patients with thrombus, multivariate analysis showed that plasma fibronectin levels were not an independent predictor of LV thrombus formation (P=.059). In multivariate analyses, only peak creatine phosphokinase (CPK) level and LV wall motion score index (WMSI) were independent predictors of thrombus formation (P=.007 and P=.0001, respectively). These results suggest the increased plasma fibronectin levels may be one of the risk factors for LV thrombus formation after AMI. However, further studies concerning the relation between plasma fibronectin levels and LV thrombus formation are necessary. PMID- 11864706 TI - Algorithms for the diagnosis of deep-vein thrombosis in patients with low clinical pretest probability. AB - The objective of this current trial was to evaluate the rate of deep-vein thrombosis (DVT) in patients after low clinical risk stratification and to evaluate the value of D-dimer and different imaging techniques in the diagnostic algorithm. A total of 99 consecutive patients were included in this prospective trial. After clinical risk assessment, D-dimer was determined. Final diagnosis was based on the results of duplex sonography, in cases of indeterminate scans on those of ascending venography. Three months after admission, follow-up investigations were performed by a telephone interview to evaluate possible further venous thromboembolism. Final diagnosis was based on the results of colour Doppler ultrasound in 92.9% and on those of venography in 7.1%. DVT was diagnosed in 2%, D-dimer was positive in 48.4%, giving a sensitivity of 100%, a specificity of 52.7% and a negative predictive value of 100%. Follow-up was possible in 89.9%--no further thromboembolic event occurred. In this specific patient group, a negative D-dimer excludes DVT and can therefore reduce the number of imaging procedures by one-half, which, on the contrary, is necessary in patients with positive D-dimer. PMID- 11864707 TI - Recurrent pulmonary embolism in a 13-year-old male homozygous for the prothrombin G20210A mutation combined with protein S deficiency and increased lipoprotein (a). AB - We report the case of a 13-year-old male presenting with recurrent symptoms of respiratory distress after a trauma of the lower limb. Pulmonary symptoms had been misinterpreted for several weeks and only marked symptoms of deep venous (DVT) and caval vein thrombosis later prompted the correct diagnosis of DVT and embolic events and subsequently a successful thrombolytic therapy. The case reported here shows a diagnostic pitfall of pulmonary embolism in an adolescent. It emphasizes the need to consider the possibility of thromboembolic events also in young children and adolescents presenting with atypical pulmonary symptoms and suffering from pulmonary diseases not responding to antibiotic therapy. In addition, although the homozygous PT A20210A gene mutation is a rare defect and its relevance as a risk factor on its own remains to be elucidated, this case suggests that a complete thrombophilia laboratory workup should be performed in young patients with a first symptomatic thromboembolic onset. PMID- 11864708 TI - Decrease of fibrinolytic activity in human endothelial cells by arsenite. AB - Blackfoot disease (BFD) is an endemic peripheral vascular occlusive disease that occurred in the southwest coast of Taiwan. It is believed that arsenic in the drinking water from artesian wells plays an important role in the development of the disease. We have previously shown that BFD patients had significant lower tissue-type plasminogen activator (t-PA) antigen level and higher plasminogen activator inhibitor, Type 1 (PAI-1) antigen level than normal controls. The purpose of this study was to investigate the effects of arsenite on the fibrinolytic and anticoagulant activities of cultured macrovascular and microvascular endothelial cells. Incubation of human microvascular endothelial cells (HMEC-1), but not human umbilical vein endothelial cells (HUVECs), with arsenite caused a decrease of t-PA mRNA level, a rise of both PAI-1 mRNA level and PAI activity. Arsenite could also inhibit the thrombomodulin (TM) mRNA expression and reduce the TM antigen level in HMEC-1. In conclusion, arsenite had a greater effect on HMEC-1 as compared to HUVECs in lowering the fibrinolytic activity and may be responsible for the reduced capacity of fibrinolysis associated with BFD. PMID- 11864709 TI - Novel alternatively spliced form of beta(3)-endonexin. AB - beta(3)-Endonexin is a binding protein to the cytoplasmic tail of beta(3) integrin and can activate alpha(IIb)beta(3) in Chinese hamster ovary (CHO) cells. Initially, two forms were identified, and only the shorter form showed the function. However, it localized mainly to the nucleus because of a nuclear localization signal (K(62)RKK). We identified two additional forms of beta(3) endonexin. One encoded 177 amino acids and was identical to the protein previously reported as a thyroid hormone receptor-binding protein. The other is a novel shortest form encoding 62 amino acids. Although the novel form lacked nuclear localization signal and was observed diffusely in the cytoplasm of transfected cells, this form did not show interaction with beta(3) integrin. Then, the ideal form as an integrin modulator was not found among these isoforms. Nevertheless, when the nuclear localization signal of the shorter form was disrupted, beta(3)-endonexin was localized near the cell surface and modulated the affinity of alpha(IIb)beta(3) more intensively. These results suggest the presence of various isoforms and the relationship between subcellular localization and integrin-activating function of beta(3)-endonexin. PMID- 11864710 TI - Adenosine increases human platelet levels of cGMP through nitric oxide: possible role in its antiaggregating effect. AB - Adenosine is an endogenous antiaggregating substance that influences the platelet responses through specific A-type receptors that activate adenylate cyclase increasing the levels of 3',5'-cyclic adenosine monophosphate (cAMP). In this study, we investigated whether adenosine can also influence the levels of 3',5' cyclic guanosine monophosphate (cGMP) and decrease the aggregating response of human platelets to adenosine-5-diphosphate (ADP) through this nucleotide. In platelet samples from healthy volunteers, we evaluated the effect of adenosine on ADP-induced aggregation and cyclic nucleotide synthesis. Some experiments were repeated in the presence of dipyridamole (inhibitor of adenosine uptake and phosphodiesterase activity), N(G)-monomethyl-L-arginine (L-NMMA, nitric synthase inhibitor), ionomycin (calcium ionophore), and ambroxol (2-amino-3,5-dibromo-N [trans-4-hydroxycyclohexyl]benzylamine, inhibitor of nitric oxide (NO)-dependent activation of guanylate cyclase). Adenosine decreased the response to ADP in a concentration-dependent way (analysis of variance, ANOVA: P<.0001): cAMP levels increased from 30.0 +/- 2.0 (control) to 46.0 +/- 3.0 pmol/10(9) platelets (in the presence of 15 mumol/l adenosine) and cGMP levels increased from 5.6 +/- 1.0 (control) to 10.9 +/- 2.0 pmol/10(9) platelets (in the presence of 15 mumol/l adenosine). Also, nucleotide levels measured at the end of aggregation were higher in platelet samples exposed to adenosine than in controls. Dipyridamole at 40 mumol/l slightly increased adenosine's effects on both nucleotides. L-NMMA blunted the effect of adenosine on cGMP both in unstimulated samples and in aggregated platelets without any effect on cAMP synthesis. Platelet exposure to L NMMA and ambroxol partially prevented adenosine's effect on ADP-induced aggregation. In conclusion, adenosine, which enhances intraplatelet cAMP levels, was determined to also cause an increase in cGMP concentrations through a mechanism that involves NO synthesis. This effect plays a direct role in the adenosine-induced antiaggregation. PMID- 11864711 TI - Snake venom disintegrin, saxatilin, inhibits platelet aggregation, human umbilical vein endothelial cell proliferation, and smooth muscle cell migration. AB - A novel disintegrin, saxatilin, was purified from Korean snake (Gloydius saxatilis) venom by means of chromatographic fractionations. We have also isolated the cDNA encoding the disintegrin using cDNA library of the snake venom gland and analyzed its complete nucleotide sequence. Saxatilin is a single-chain polypeptide composed of 73 amino acids including 12 cysteines as well as the tripeptide sequence Arg-Gly-Asp (RGD), a proposed recognition site of adhesive proteins. Molecular mass of saxatilin was determined to be 7712 Da by matrix assisted laser desorption ionization mass spectrometry. Saxatilin inhibits glycoprotein (GP) IIb-IIIa binding to immobilized fibrinogen with IC(50) of 2.0 nM and ADP-induced platelet aggregation with IC(50) of 127 nM, respectively. The snake venom disintegrin also significantly suppresses basic fibroblast growth factor-induced human umbilical vein endothelial cell (HUVEC) proliferation, but has little effect on normal growth of the cell. Interaction of human umbilical vein cell to immobilized vitronectin is also inhibited by binding of saxatilin to alpha(v)beta(3) integrin. Adhesion of smooth muscle cells (SMCs) to vitronectin as well as vitronectin-induced migration of the cells was strongly inhibited by saxatilin. Several lines of experimental evidence suggest potential use of saxatilin for development of therapeutic agents. PMID- 11864712 TI - Molecular basis for different ability of low-density and high-density lipoproteins to support activity of the intrinsic Xase complex. PMID- 11864713 TI - Role of each Asn-linked glycan in the anticoagulant activity of human protein C inhibitor. AB - The N-glycosylation site mutants of human protein C inhibitor (PCI; N230S, N243Q, N319Q, N230S/N243Q, and N230S/N319Q) were prepared by amino acid replacement of the asparagine residue with a serine or glutamine residue using site-directed mutagenesis and expressed in the baculovirus/insect cell expression system. To examine the importance of each Asn-linked glycan in the activity of PCI, we compared wtPCI with the mutants of N-glycosylation site(s) in terms of the procoagulant protease-inhibitory and anticoagulant activities. The inhibitory activities of N230S, N319Q, and N230S/N319Q toward human thrombin and plasma kallikrein were significantly increased compared with wtPCI, but those of N243Q and N230S/N243Q were reduced. The inhibitory activity of N230S toward human plasma coagulation was significantly increased compared with wtPCI, and that of N230S/N319Q was also significantly increased compared with N319Q. Furthermore, the procoagulant protease-inhibitory and anticoagulant activities of N230S/N319Q (glycosylated on Asn243 only) compared favorably with those of N230S, and both of the mutants possessed highest activities in the purified mutants. These results suggest that the Asn243-linked glycan in PCI molecule possesses critical roles for its anticoagulant activity in the circulation, and the Asn230-linked glycan down-regulates the activity of PCI. PMID- 11864715 TI - Psychological heterogeneity in AD/HD--a dual pathway model of behaviour and cognition. AB - Psychological accounts have characterised attention-deficit/hyperactivity disorder (AD/HD) as either a neuro-cognitive disorder of regulation or a motivational style. Poor inhibitory control is thought to underpin AD/HD children's dysregulation while delay aversion is a dominant characteristic of their motivational style. A recent 'head to head' study of these two accounts suggest that delay aversion and poor inhibitory control are independent co existing characteristics of AD/HD (combined type). In the present paper we build on these findings to propose a dual pathway model of AD/HD that recognises two quite distinct sub-types of the disorder. In one AD/HD is the result of the dysregulation of action and thought resulting from poor inhibitory control associated with the meso-cortical branch of the dopamine system projecting in the cortical control centres (e.g. pre-frontal cortex). In the other AD/HD is a motivational style characterised by an altered delay of reward gradient linked to the meso-limbic dopamine branch associated with the reward circuits (e.g. nucleus accumbens). The two pathways are further distinguished at the levels of symptoms, cognitive and motivation profiles and genetic and non-genetic origins. PMID- 11864714 TI - How specific is a deficit of executive functioning for attention deficit/hyperactivity disorder? AB - A selective review of research in the executive functioning (EF) is given for attention deficit hyperactivity disorder (ADHD), oppositional defiant disorder (ODD), conduct disorder (CD), higher functioning autism (HFA) and Tourette syndrome. The review is restricted due to changes in the classification of the disorder in recent years and secondly the heterogeneity of EF is restricted to five key areas of concern, inhibition, set shifting, working memory, planning, and fluency. The review makes clear that there are strong differences between child psychopathological groups and controls on these EFs. However, future research will be needed to identify an EF deficit or profile, which is specific for these disorders. PMID- 11864716 TI - Attention-deficit/hyperactivity disorder (ADHD) behaviour explained by dysfunctioning reinforcement and extinction processes. AB - Inattentiveness, overactivity and impulsiveness are presently regarded as the main clinical symptoms of attention-deficit/hyperactivity disorder (ADHD). Inattention is, however, a characteristic of most psychiatric disorders. It is argued that the ADHD Inattentive subtype may have heterogeneous origins and be qualitatively different from the ADHD Hyperactive/Impulsive subtype. At the neurobiological level, ADHD symptoms may to a large extent be caused by a dysfunctioning dopamine system: A dysfunctioning meso-limbo-cortical dopamine branch will produce altered reinforcement and extinction processes, on a behavioural level giving rise to deficient sustained attention, hyperactivity, motor and cognitive impulsiveness. A dysfunctioning nigro-striatal dopamine branch will cause 'extrapyramidal' symptoms. Our model disentangles the behaviours usually explained by 'executive functions' into cognitive impulsiveness, motor impulsiveness and deficient motor control. The various dopaminergic branches may not be equally dysfunctional in all individuals with ADHD. Etiologically, dopamine dysfunctioning will probably mainly be genetically determined while sometimes be induced by environmental factors like drugs of abuse or pollutants, which may explain geographical differences in prevalence rates. PMID- 11864717 TI - The dynamic approach to neurodevelopmental psychiatric disorders: use of fMRI combined with neuropsychology to elucidate the dynamics of psychiatric disorders, exemplified in ADHD and schizophrenia. AB - The paper discusses the application of fMRI in combination with neuropsychology to neurodevelopmental psychiatric disorders, exemplified on the case of attention deficit hyperactivity disorder (ADHD) in comparison with schizophrenia. The view is presented that ADHD, rather than being a compound of unrelated co-existing deficits, is a pervasive disorder of impulsiveness, which manifests at the motor, emotional, social and cognitive domain. Neuropsychology needs to refine the psychological measurements of these impulsivity symptoms and, in combination with fMRI, provide new insights into the interrelationship between brain and dysfunction and its bi-directional causalities. The suitability of the dynamic technique of functional MRI to assess the dynamic nature of developmental neuropsychiatric disorders is discussed. Brain activation can inform about strategy and compensatory mechanisms at a neuroanatomical level, which are not observable at a psychological level, providing insight into the underlying neurocognitive mechanisms of psychiatric disorders. Data are presented and discussed on opposing neurocognitive activation patterns for patients with ADHD and those with schizophrenia while performing a stop task. Comparisons between patient groups will be essential to address the specificity of neurocognitive mechanisms corresponding to specific neurodevelopmental psychiatric disorders. PMID- 11864718 TI - The dopamine transporter: relevance to attention deficit hyperactivity disorder (ADHD). AB - The dopamine transporter is elevated in adults with attention deficit hyperactivity disorder (ADHD) compared with healthy controls [Lancet 354 (1999) 2132]. The findings have been confirmed by others in a different population using a different probe for the dopamine transporter. Notwithstanding the need to confirm these findings in a multi-center trial, several hypotheses are presented to account for these observations. A premise that elevated transporter levels result from medication is not supported by current data. Other possibilities, including hypertrophy of dopamine neuronal terminals in the striatum, dysfunctional regulation of dopamine or dopamine receptors, or anomalies in the dopamine transporter gene are presented as hypotheses. The feasibility of exploring these mechanisms in animal models or in human subjects is explored. PMID- 11864719 TI - Dopamine dysfunction in AD/HD: integrating clinical and basic neuroscience research. AB - There is strong evidence that the catecholamines dopamine and norepinephrine are both important in the pathophysiology of ADHD, as well as in the mechanism of therapeutic action of stimulant drugs. Due to the known effects of stimulants in blocking reuptake of catecholamines and (in the case of D-amphetamine) facilitating their release, it has traditionally been believed that the stimulants compensate for catecholamine deficiency in ADHD. More recently, however, alternate hypotheses of a hyperdopaminergic and/or hyper-noradrenergic state in ADHD have been suggested. This paper will be limited to a review of the evidence for involvement of dopamine in mediating behavioral and cognitive symptoms and response to stimulants in ADHD, with implications for possible mechanisms. PMID- 11864720 TI - Pharmacokinetic and pharmacodynamic properties of stimulants: implications for the design of new treatments for ADHD. AB - In the USA, the stimulant drug methylphenidate (MPH) is used to treat a large number (2 million or more per year) of children with Attention Deficit Hyperactivity Disorder (ADHD). Although the US FDA approved MPH in the 1960s, the pharmacokinetic (PK) properties of serum concentrations of MPH in children with ADHD were not described until the 1980s, and then in only a few cases. Recently, information from drug development programs have increased our knowledge about the serum PK and some pharmacodynamic (PD) characteristics of MPH in ADHD children, and studies based on positron emission tomograpy (PET) in adult volunteers have provided new knowledge about the PK properties of MPH at the primary site of action in the brain. We will review these two topics and use this new information to evaluate the mechanisms of action of MPH. PMID- 11864721 TI - Methylphenidate elevates resting dopamine which lowers the impulse-triggered release of dopamine: a hypothesis. AB - How do 'stimulants' reduce hyperactivity in children and adults? How can drugs which raise extracellular dopamine result in psychomotor slowing of hyperactive children when dopamine is known to enhance motor activity, such as in Parkinson's disease? In summary, the hypothesis for the anti-hyperactivity effects of the stimulants is as follows: during normal nerve activity, extracellular dopamine levels transiently rise 60-fold. At low therapeutic doses (0.2-0.5 mg/kg) to treat attention-deficit hyperactivity disorder, stimulant drugs such as methylphenidate and dextroamphetamine reduce locomotion in both humans and animals. The drugs raise resting extracellular levels of dopamine several-fold, but reduce the extent to which dopamine is released with nerve impulses, compared to the impulse-associated release in the absence of the drug. This relatively reduced amplitude of impulse-associated dopamine would result in less activation of post-synaptic dopamine receptors which drive psychomotor activity. At higher doses, stimulants produce generalized stimulation of the nervous system, as a result of the very high concentrations of extracellular dopamine at rest, and the markedly increased release of dopamine with nerve impulses. These high levels of resting and pulsatile dopamine cause widespread stimulation of post-synaptic dopamine receptors, overcoming any concomitant presynaptic inhibition of dopamine release. PMID- 11864722 TI - Psychiatric pharmacogenetics: personalizing psychostimulant therapy in attention deficit/hyperactivity disorder. AB - There is a substantial amount of variation in response and adverse drug reactions to psychostimulant therapy in attention-deficit/hyperactivity disorder (ADHD). Psychiatric pharmacogenetics is a rapidly developing field, which can be applied to identify genetic predictors of this variability in outcome to psychostimulant medications. This article will briefly review ADHD and its pharmacotherapy. This will be followed by an overview of the pharmacokinetics and pharmacodynamics of methylphenidate, the most commonly used psychostimulant in the US. Then the field of psychiatric pharmacogenetics will be introduced and its methodology will be described. This will be followed by a discussion about how pharmacogenetics can be applied to children afflicted with ADHD. The future of psychiatric pharmacogenetics will then be presented with an emphasis being placed on developing prospects that will ensure the continued advancement of this field. PMID- 11864723 TI - A transmission disequilibrium test of the Ser9/Gly dopamine D3 receptor gene polymorphism in adult attention-deficit hyperactivity disorder. AB - Convincing data support the hypothesis that genetic factors are involved in the etiology of attention-deficit hyperactivity disorder (ADHD). Various lines of evidence have shown that the dopamine system plays a crucial role in the pathophysiology of ADHD. The dopamine D3 receptor gene (DRD3) represents a promising candidate to examine in ADHD. Animal studies have shown that DRD3 mRNA is highly expressed in the ventral striatum suggesting an involvement of this receptor in the control of motor behaviour. Manipulation of DRD3 in rodents has led to a mouse model with nonfunctional D3 receptors that displays hyperactive behaviour in various environmental conditions. Furthermore, administration of 7 OH-DPAT, a dopaminergic agonist that binds preferentially to D3 receptors exerts an inhibitory effect on locomotor activity while D3 antagonists induce hyperactivity. Among various polymorphisms described for DRD3, the BalI polymorphism is most interesting because it codes for an aminoacid substitution in the N-terminus of the receptor. The receptor products of the two alleles (Ser/Gly) exhibit differential affinity for dopamine. To determine if DRD3 Ser9/Gly is involved in the susceptibility to ADHD we genotyped 39 adults with ADHD and their respective parents (trios). Adult ADHD represents a promising phenotype for studying the genetic component of the disorder. In fact, a recent family study has shown that relatives of adult ADHD patients have a higher rate of ADHD compared to relatives of children with ADHD suggesting a stronger genetic component for the adult version. The results of genotyping in the 39 trios analyzed with the transmission disequilibrium test showed no excess of transmission for DRD3 MscI/BalI alleles (chi(2) = 0.360; df = 1; P = 0.54). This result, although from a relatively small sample, indicates that it is unlikely that DRD3 is playing a major role in the etiology of ADHD in our sample. PMID- 11864724 TI - Dopamine may be 'hyper' with respect to noradrenaline metabolism, but 'hypo' with respect to serotonin metabolism in children with attention-deficit hyperactivity disorder. AB - Noradrenaline: Hechtman (J Psychiat Neurosci 1994;19:193) argued for a role for frontal dopamine (DA) and noradrenaline (NA) in ADHD, where Oades (Prog Neurobiol 1987;29:365) has described lateralised functional impairments. Mechanisms (e.g. via alpha-2 sites) for stimulating low NA activity in ADHD children (J Am Acad Child Adolesc Psychiatry 1997;36:1688) in order to promote interactions with mesocortical DA have been discussed (J Psychopharmacology 1997;11:151; Psychiatr Res 1994;52:305). We described with indicators of overall transmitter metabolism (monoamines, metabolites in 24 h urine samples (Behav Brain Res 1997;88:95)) significantly lower utilisation ratios (MHPG/NA) in ADHD children with respect to healthy controls. Interestingly, a comparison of between catecholamine levels (DA/NA) showed a correlation with the conditioned blocking measure of selective attention recorded at the time of collection. This measure was negatively associated with blocking in controls. These results are consistent with reports of lower DOPEG and increased DOPAC in ADHD urine (J Child Adolesc Psychopharmacol 1996;6:63) and indicate that the relatively hyperactive DA versus NA systems may have functional consequences. Serotonin: the relevance for ADHD of an association of impulsivity with low serotonin (5-HT) metabolism (Behav Brain Sci 1986;9:319) has long been played down. Yet, some symptoms have been related to CSF measures of the metabolite 5-HIAA, and in particular the HVA/5-HIAA ratio has been reported to correlate with ratings of activity (Psychiatr Res 1994;52:305). We find that while urinary measures of 5-HIAA are somewhat higher, the ratio HVA/5 HIAA is markedly lower in ADHD children versus controls. In these ADHD children 5 HIAA levels were negatively related to d-prime measures in a continuous performance task (CPTax), and the HVA/5-HIAA was negatively associated with conditioned blocking. These results suggest a relatively low DA versus 5-HT activity may have functional consequences, albeit in a subgroup of ADHD. This is consistent with drug-induced prolactin changes reported by Verbaten et al. (Eur Child Adolesc Psychiatry 1999;8:30). PMID- 11864725 TI - The contribution of comparative studies in inbred strains of mice to the understanding of the hyperactive phenotype. AB - Attention-deficit hyperactivity disorder (ADHD) is a highly prevalent childhood psychiatric disorder characterized by impaired attention, excessive motor activity and impulsivity. Converging evidence, suggests a primary role of disturbances in brain dopamine (DA) transmission and a role of genetic factors in its pathology. Inbred provide a well-defined and stable genotype for analysis. C57BL/6 (C57) and DBA/2 (DBA) mice are amongst the most studied inbred strains in the behavioral pharmacology of DA, and they differ in several parameters of the DA system that relate directly to behavioral differences. These strains also exhibit several qualitatively different behavior patterns that rely on separate DA networks (e.g. mesoaccumbens vs. nigrostriatal) and on different modes of inheritance. C57 mice are good learners in most tasks also involving associative learning but are totally unable to learn active avoidance although being very active. Moreover, C57 mice show greater novelty-induced locomotor activity than DBA, which is modulated strongly by DA neurons in the ventral tegmental area (VTA) region. Pharmacological studies also indicate a facilitated mesoaccumbens DA transmission in C57 mice when compared to DBAs. Increased density of D2 autoreceptors located on VTA neurons, and lower D2 postsynaptic receptors in the NAS were observed in DBA relative to C57. Activation of D2 autoreceptors inhibits impluse flow, synthesis, and release rates of DA neurons. As would be predicted from their higher D2 autoreceptor: DBA compared to C57 mice show reduced DA synthesis and release within the mesoaccumbens DA system when challenged with DA direct agonists. However, DBA mice are by fare more susceptible than C57s to stress-induced enhanced mesoaccumbens DA release and in stressful situation, they show sustained active behavioral responses whilst C57 adopt extremely passive responses (behavioral despair). Finally, chronic or repeated stress promote opposite adaptation of VTA DA autoreceptors in the two strains and render the hypoactive DBAs as active as the C57 mice. These results indicate that a complex interaction between genetic and environmental factors controls, mesoaccumbens DA functioning and hyperactive phenotype. PMID- 11864726 TI - Environmental factors during postnatal period modify activity and non-selective attention in the Naples High-Excitability rat. AB - The involvement of epigenetic factors in the phenotypic expression of the neural systems underlying activity and attentive processes has been investigated in an animal model of hyperactivity and attention-deficit, the Naples High-Excitability rat (NHE). To this aim, male NHE pups have been reared in small (four) or normal litter size (nine) during the first 4 weeks of postnatal life. Both groups underwent a differential handling procedure occurring once, twice or four times a week. After weaning (28 days), rats were housed in groups of two and tested as young adults for activity and non-selective attention in a spatial novelty situation for three consecutive tests at 24-h intervals. The behaviour was videotaped and analysed off line for the frequency of corner crossings and rearings and duration of rearings. The results indicate that the increased maternal care and high fat diet induced by the small litter size produced long lasting effects on activity and duration of rearing episodes that indexes non selective attention. These effects were complex as differential handling was beneficial only at low stimulation level. Thus, these findings suggest that epigenetic factors acting during critical periods of post-natal development may interact with genetic determinants that in turn influence the maturation of the neural systems controlling activity, orienting and scanning time. PMID- 11864727 TI - Peculiar response of adolescent mice to acute and chronic stress and to amphetamine: evidence of sex differences. AB - Although final brain size and the number of available neurons and axons appear to be established early in infancy, plasticity of the brain continues during adolescence through an integrated process of overproduction and elimination of synapses and receptors. In addition, hormonal levels change dramatically during this period, as a result of the onset of puberty. This age-specific condition has been suggested to serve as a permissive factor for the emergence of a number of early-onset neuropsychiatric disorders, including schizophrenia, attention deficit hyperactivity disorder (ADHD), and perhaps substance abuse. However, relatively few investigations have focused on animal models of this developmental phase. The periadolescent rodent (similar30-45-day-old), has been proposed as a useful model. Periadolescent rats and mice are generally associated with a peculiar behavioral profile, consisting of basal hyperactivity, high attraction towards novel stimuli and a marked involvement in affiliative and playful behaviors. Moreover, a unique profile of psychopharmacological responsivity characterizes rodents around this age. Recent experiments by our group investigated age-related discontinuities in the response of the hypothalamic pituitary-adrenal axis (HPA) to both stress and psychostimulants. The latter are often administered as therapeutic drugs to children with ADHD, which have been also associated with an impaired response to stress and abnormalities in HPA axis function. Indeed, an altered functioning of the HPA axis has been proposed as a possible risk factor and a potential marker for such a behavioral vulnerability. Animals were studied at adulthood (> pnd 70) or during periadolescence. Experiment I characterized basal corticosterone (CORT) levels in naive mice kept undisturbed in standard social conditions from weaning to sacrifice. Periadolescent male mice showed higher basal CORT levels than adult subjects, suggesting that the set up of the HPA axis is physiologically elevated during adolescence. In experiment II, we investigated age-related differences in the response to both acute and chronic stress conditions. Periadolescent and adult mice were housed either in a standard (three animals per cage) or in a crowding condition (nine animals per cage). The latter has been indeed reported to potentiate the subsequent reaction to acute stress in adult rodents. At the end of this period and following 24 h individual housing, mice were injected with either saline (SAL) or a standard amphetamine (AMPH) dose (2 mg/kg), and faced with a mild acute psychological stress, namely removal of sawdust from the home cage. Important sex differences emerged in animals of the two ages. Periadolescent females showed a reduced CORT response to acute stress. Within the adult male group, the chronic crowding condition produced a prominent potentiation of CORT response to the acute stress challenge. Conversely, this profile was not evidenced in periadolescents. These results indicate a strong role for gender and social variables in the response of periadolescent subjects to the various aspects of stress. As for AMPH effects, in the absence of significant changes in adult subjects, the drug produced a marked CORT release in periadolescent mice. A better understanding of neuroendocrine-related AMPH effects as a function of social and environmental risk factors during adolescence, might deepen our knowledge on the neurobiological bases of genetically determined neuropsichiatric disorders and possibly improve the therapeutical efficacy of psychostimulant drugs. PMID- 11864728 TI - Selective inhibition of neuronal nitric oxide synthesis reduces hyperactivity and increases non-selective attention in the Naples High-Excitability rat. AB - The involvement of neuron-derived NO in the process of orienting and scanning times (non-selective attention: NSA) towards environmental stimuli has been investigated in the Naples High-Excitability rat (NHE), a putative animal model of Hyperactivity and Attention Deficit (ADHD). To this aim, orienting and scanning times have been monitored by the frequency and duration of rearing episodes, respectively. Adult male NHE rats were tested in a novelty situation (Lat-maze) for 30 min following single or repeated injections of the non competitive inhibitor 7-Nitroindazole (7-NINA) of the neuronal isoform of the enzyme nitric oxide synthase (n-NOS). In the acute experiments, rats received a single injection of 7-NINA (1 mg/kg) intraperitonealy in a saline vehicle (exp. 1, fast release) or subcutaneously in a lipid carrier, dimethyl sulfoxide (DMSO; exp. 2, slow release) or the vehicles alone as controls 30 min before testing. In the repeated injection experiments, rats received a subcutaneus injection of 1 mg/kg in DMSO or DMSO alone daily for 14 days, and tested 24 h after the last injection (exp. 3, slow release). The results showed a significant differential effect of the drug that was dependent on the release rate, i.p. saline-diluted 7 NINA increased the duration of individual rearing episodes whereas, both single and repeated subcutaneous DMSO-carried 7-NINA exerted an opposite effect. Thus, selective inhibition of n-NOS by an allosteric inhibitor that increases arginine availability without displacing the inhibitor from n-NOS, strengthens the hypothesized role of NO in NSA. These findings may shed light on the mechanism of action of drug treatment of and be useful in the treatment of ADHD in children. PMID- 11864729 TI - Impaired metabolic capacity in the perirhinal and posterior parietal cortex lead to dissociation between attentional, motivational and spatial components of exploration in the Naples High-Excitability rat. AB - This study aimed at investigating the neural substrates of spatial and non spatial Behavioural components of exploration to novelty by a neurogenetic approach. Thus, functional imaging and Behavioural analysis were carried out in the Naples High-Excitability (NHE) rats, a model of hyperactivity and attention deficit. Quantitative cytochrome oxidase (C.O.) histochemistry was used to measure the basal metabolic capacity of different forebrain structures. In parallel experiments, exploration in an 8-arm radial maze (Olton-maze) with extra maze cues was used to measure attentional, motivational and spatial components of Behaviour after feeding rats' ad-libitum or at a reduced diet. Functional imaging analysis: brains from naive rats were stained for quantitative C.O. histochemistry along with standards. NHE rats showed lower C.O. activity in perirhinal and posterior-parietal cortex (all layers) and cortical amygdala, and greater activity in entorhinal cortex (superficial layers). The outer granular cell layer of the dentate gyrus had greater activity in NHE. Behavioural analysis: at low and high motivational level, maze exploration was reinforced during shaping throughout and then only a single arm. The Behaviour was monitored by a CCD camera and videotaped. (i) There was no line difference in working memory during non reinforced maze exploration, independent of the motivational level; (ii) during shaping with all baited arms, there was no line difference in working memory, but NHE rats showed a very low or lower food consumption at low and high motivational level, respectively; (iii) rats showed a higher working memory in finding the single baited arm at high motivational level; (iv) NHE rats paid little attention towards reinforcement upon visiting the baited arm only at low motivational level. Thus, Behavioural and functional neuroimaging analysis suggests the neural substrates of spatial and non-spatial components of exploration to be underlined by different network operations in the neocortical and limbic cortices in the NHE rat lines. Therefore, they appear as an useful tool to the understanding of Attention-Deficit Hyperactivity Disorder (ADHD) in children. PMID- 11864730 TI - Activity, non-selective attention and emotionality in dopamine D2/D3 receptor knock-out mice. AB - In order to assess the role of dopamine (DA) D2 and D3 receptors in the modulation of behaviour, we analysed exploration in a spatial novelty in mouse model systems. Genetically engineered mice mutants have been used that carry normal, partial or no expression of D2R, D3R, or both D2R/D3R (double mutants) DA receptor subtypes. Adult male mice were exposed for 30 min to a Late-maze. The behaviour was analysed for indices of activity, orienting (rearing frequency), scanning times (rearing duration) and defecation score (emotionality). D2R - / - and + / - as well as the D2R/D3R double homozygous mutants were less active than wild-type (WT) controls in travelled distance. In contrast D3R + / - were more active than WT mice in the first part of the test. As to orienting frequency, the D2R - / - were less active than WT during the entire test-period, whereas the D2 + / - mutants were less active than WT only in the second part of the test. Moreover, the D3R - / - and + / - mutants showed less and more rearing frequency than WT, respectively, during the entire test. Finally, the D2/D3R - / - double mutants were also less active than WT during the entire test period. As to scanning times, D2R + / - and - / - mutants were higher than WT during the entire test or only in the second part, respectively. The D3R + / - and - / - were not different from WT, whereas the D2/D3R - / - double mutants showed shorter scanning times only in the first part of the test. As to emotionality index, the defecation score, was lower only in D3R + / - mutants. Thus, the dopamine D2 and D3 receptor subtypes appear to be differentially involved in the modulation of activity, orienting and scanning phases of attention. Lastly double mutation experiments reveal an interaction between D2R and D3R with the former prevailing on the latter. PMID- 11864731 TI - Intrinsic membrane properties and synaptic inputs regulating the firing activity of the dopamine neurons. AB - Dopamine (DA) neurones of the ventral mesencephalon are involved in the control of reward related behaviour, cognitive functions and motor performances, and provide a critical site of action for major categories of neuropsychiatric drugs, such as antipsychotic agents, dependence producing drugs and anti-Parkinson medication. The midbrain DA neurones are mainly located in the substantia nigra pars compacta (SNPC) and the ventral tegmental area (VTA). Intrinsic membrane properties regulate the activity of these neurones. In fact, they possess several conductances that allow them to fire in a slow pacemaker-like mode. The internal set of membrane currents interact with afferent synaptic inputs which, especially in in vivo conditions, contribute to accelerate or decelerate the firing activity of the cells in accordance with the necessity to optimise the release of dopamine in the terminal fields. In particular, discrete excitatory and inhibitory inputs transform the firing from a low regular into a bursting pattern. The bursting activity promotes dopamine release being very important in cognition and motor performances. In the present paper we review electrophysiological data regarding the role of glutamatergic and cholinergic and GABAergic afferent inputs in regulating the midbrain DAergic neuronal activity. PMID- 11864732 TI - A rostro-caudal dissociation in the dorsal and ventral striatum of the juvenile SHR suggests an anterior hypo- and a posterior hyperfunctioning mesocorticolimbic system. AB - Functional molecular neuroimaging techniques have been applied to the study of the neural substrates of Attention-Deficit Hyperactivity Disorder (ADHD) in an animal model, the juvenile SHR rat. They include quantitative receptor autoradiography and immunocytochemistry for neuronal markers such as Ca2+/Calmodulin Dependent Kinase II (CaMKII) and transcription factors. Multiple evidence emerges for a rostro caudal dissociation within the dorsal (DS) and ventral striatum (VS) (n. accumbens) and olfactory tubercle (OT). It consists in (i) a higher density of dopamine (DA) D-1/D-5 receptor binding sites in a discrete segment of the anterior forebrain that comprises the DS, VS and OT, (ii) a lower density of DA D-2/D-3 autoreceptors in the caudal portion of the n. accumbens shell subterritory, (iii) a reduced number of CaMKII and c-FOS positive elements only in the anterior portion of DS and VS (iv) reversal by repeated injections of methylphenidate (MP) (3 mg/kg, 14 days) with 'downregulation' in SHR and 'up-regulation' in the WKY control rats of DS and VS of DA D-1/D-5 receptors. Thus, under basal conditions the mesocorticolimbic (MCL) DA system appears to be hyperfunctioning rather than hypofunctioning, as demonstrated (i) by subsensitivity of presynaptic D-3 autoreceptors and (ii) by phasic inhibition of MCL activity induced by acute blockade of endocannabinoid reuptake using AM404. Following MP treatment, the hyperfunctioning MCL DA system turns into a hypofunctioning one, as earlier suggested by Solanto. Since the target neurons of MCL fibers seem to be uncoupled to D-1 receptors, the medium spiny GABA neurons projecting to the ventral pallidum and ventral tegmental area (VTA) exert a weak feedback inhibition on the neurons of origin of MCL system. Therefore, MCL neurons maintain a high basal activity with consequences on the cortico-striato pallido-thalamo-cortical system and amygdala complex through the 'extended amygdala system'. While the former explains the attention, motivation and activity alterations of this rat model of ADHD, the latter explains the emotional symptoms of the syndrome. It remains to be ascertained the starting point in the network leading eventually to the segmental defect as well as its significance in humans. PMID- 11864733 TI - A morphometric evidence for a hyperfunctioning mesolimbic system in an animal model of ADHD. AB - The hyperfunctioning dopamine hypothesis in the mesocorticolimbic (MCL) system has been addressed by a neurogenetic approach in model systems. Thus, a morphometric analysis was carried out on neurons of origin of Substantia Nigra (SN) and Ventral Tegmental Area (VTA) dopamine systems of the Naples High Excitability (NHE), Low-Excitability (NLE) and control lines. Male adult rats were tested in a spatial novelty for indices of activity and non-selective attention. Mesencephalic coronal sections were processed for tyrosine hydroxylase (TH) immunohistochemistry and cytochromoxidase (C.O.) histochemistry. Image analysis in the rostro-caudal plane showed (i) a higher neuron size of TH+ elements in the VTA of NHE and NLE, across the entire structure in the NHE, and only in the middle portion in the NLE; (ii) a higher expression of TH in the neuropil of the VTA in NHE; (iii) a lower C.O. activity in both NLE and NHE; (iv) no differences in the SN. The larger neuron size in both NHE and NLE rats as compared with control rats, along with higher TH expression mainly in the NHE, in absence of any relevant alteration in the SN, reveals an unbalance between the two dopamine systems and a subsequent alteration in limbic (reward, motivation, sustained attention) functions. The decreased C.O. activity might be due to reduced feedback inhibition by striatal GABA neurons and interneurons leading to increased DA neuron firing. In conclusion, the increased behavioral activity and impaired attention observed in the NHE rats are associated to hyperfunctioning MCL system in this genetic model of Attention-Deficit Hyperactivity Disorder (ADHD). PMID- 11864734 TI - Hypodopaminergic and hypernoradrenergic activity in prefrontal cortex slices of an animal model for attention-deficit hyperactivity disorder--the spontaneously hypertensive rat. AB - Evidence supports dysfunction of dopaminergic and noradrenergic systems in patients with attention-deficit hyperactivity disorder (ADHD). Noradrenergic and dopaminergic systems exert distinct modulatory actions on the transfer of information through neural circuits that connect functionally distinct cortical areas with separate striatal regions and remain segregated in parallel striato pallidal-thalamic and striato-substantia nigra pars reticulata-thalamic pathways. Prefrontal cortex performance is maximal at moderate stimulation of postsynaptic dopaminergic and noradrenergic receptors, and is reduced by either higher or lower levels of receptor stimulation. Spontaneously hypertensive rats (SHR) are generally considered to be a suitable genetic model for ADHD, since they display hyperactivity, impulsivity, poor stability of performance, impaired ability to withhold responses and poorly sustained attention, when compared with their normotensive Wistar-Kyoto (WKY) control rats. Evidence suggests that terminals of mesocortical, mesolimbic and nigrostriatal dopaminergic neurons of SHR release less dopamine in response to electrical stimulation and/or depolarization as a result of exposure to high extracellular K+ concentrations, than WKY. Vesicular storage of dopamine was suggested to be impaired in SHR, causing leakage of dopamine into the cytoplasm and increased d-amphetamine-induced transporter mediated release. While electrically stimulated release of dopamine appears to be decreased in prefrontal cortex of SHR suggesting hypodopaminergic function, autoreceptor-mediated inhibition of norepinephrine release appears to be impaired in SHR, suggesting that noradrenergic function may be poorly regulated in the prefrontal cortex of the SHR. These findings are consistent with the hypothesis that the behavioral disturbances of ADHD are the result of an imbalance between noradrenergic and dopaminergic systems in the prefrontal cortex, with inhibitory dopaminergic activity being decreased and noradrenergic activity increased relative to controls. PMID- 11864735 TI - Changes in the second messenger cyclic AMP during development may underlie motoric symptoms in attention deficit/hyperactivity disorder (ADHD). AB - The transitions that occur in the ascending dopamine systems between childhood and adulthood parallel the emergence, course, and severity of attention-deficit hyperactivity disorder (ADHD) symptoms. Behaviorally, rats are more active in open field during periadolescence, and activity levels decline by 50% in males by adulthood. This peak in behavior parallels a transient overproduction in D1 and D2 dopamine receptors that occurs at puberty in rat striatum (STR) and prefrontal cortex (PFC), followed by a decline in receptor density into adulthood. While tempting to speculate that receptor density plays a role in the waning of ADHD symptoms, receptor overproduction does not occur in the nucleus accumbens (NA), which demonstrates only a modest rise in receptor density (10-20%). Given the importance of the accumbens in locomotor activity, an alternative explanation for increased activity was sought. The second messenger system cyclic adenosine monophosphate (cAMP) has classically been associated with dopamine receptors. The results of these studies demonstrate that cAMP accumulation in the accumbens and the STR parallel the observed rise and fall in activity levels in rats. At puberty, basal cAMP levels are 35% higher relative to adulthood in male accumbens, while a modest 7% change was observed in STR. Forskolin-stimulated cAMP was 240-300% higher in STR and accumbens at puberty before declining with maturation. These findings suggest that, the adolescent dopamine system has a much higher 'tone' relative to adults. However, pharmacological responsiveness of cAMP to D1 or D2 stimulation demonstrates an overall blunted response during puberty relative to adulthood. This finding is consistent with a hyposensitivity to stress and pharmacological agents at puberty in animals that are in a hyperdopaminergic state. These findings of combined elevated cAMP accumulation and reduced cAMP sensitivity during adolescence have clinical implications for hypothesized mechanism and course of ADHD and its treatment. The maturational decline in cAMP activity may explain why this disorder recedes, while, simultaneously cAMP becomes more responsive to D1 and D2 receptor stimulation in adulthood. PMID- 11864736 TI - Regulation of dopamine quantal size in midbrain and hippocampal neurons. AB - Since the pioneering work of Bernard Katz and his colleagues decades ago, neurotransmitter quantal size (defined as the number of neurotransmitter molecules released by a single synaptic vesicle during exocytosis) is often modeled as invariant. This assumption had tremendous implications for basic research on synaptic plasticity. For instance, it focused attention on the postsynaptic rather than the presynaptic component in studies of learning and memory (the field of long-term potentiation comes to mind as the best example). Furthermore, this assumption somehow 'spilled over' onto studies of monoamine neurotransmitters, which apparently use diffusion and slow action to exert their modulatory effects, in contrast to the fast acting neurotransmitters studied by Katz. Consequently, research on dopamine-related diseases (e.g. psychotic and movement disorders) did not pay as much attention to presynaptic mechanisms that regulate dopamine release, as to postsynaptic receptor action. Part of the problem, of course, has been the lack of technology to directly measure quanta from presynaptic sites and the obligatory reliance on measurements of miniature postsynaptic potentials (minis) for reaching conclusions about presynaptic quantal events. Due to the introduction of the carbon fiber amperometric microelectrode in tissue electrophysiology, initially by Francois Gonon (University of Bordeaux) and then by Mark Wightman (University of North Carolina), we were able to directly measure dopamine quanta from neurites of cultured midbrain dopamine neurons by amperometry. This was the first approach to provide direct measurement of the number of molecules and kinetics of presynaptic quantal release from CNS neuronal terminals. The interventions altering dopamine quantal size are so far the following. (1) Alteration of neurotransmitter synthesis--an increase of cytosolic dopamine availability (e.g. by exposure to L DOPA) increases quantal size and a decrease of cytosolic dopamine by D2 autoreceptor activation (by quinpirole) decreases quantal size. (2) Modulation of vesicle transmitter transporter activity--overexpression of the neuronal vesicular monoamine transporter VMAT2 increases dopamine quantal size. The reduction or elimination of VMAT2 protein in mice significantly hampers or eliminates monoamine release. (3) Reuptake blockade--cocaine and amfonelic acid are dopamine reuptake blockers which reduce quantal size independently of D2 related effects. (4) Changes in transvesicular pH gradient-neuronal stimulation apparently leads to vesicular acidification via the activation of chloride channels on the vesicular membrane and increased quantal size. (5) Fusion pore kinetics--a vesicle undergoing exocytosis may discharge only part of its neurotransmitter content before recycling. Plasticity of the fusion pore shape may, therefore, be a crucial determinant of quantal size. Other possible sources of variability in quantal size are altered transmitter degranulation and changes in synaptic vesicle volume. We suggest that plasticity in dopamine quantal seems likely to be involved in both normal synaptic modification and disease states. PMID- 11864737 TI - Measles virus matrix protein is not cotransported with the viral glycoproteins but requires virus infection for efficient surface targeting. AB - As we have shown earlier, the measles virus (MV) glycoproteins H and F are expressed on both, the apical and the basolateral membrane of polarized Madin Darby canine kidney cells. In contrast to the glycoproteins, we found the viral matrix protein (M) to accumulate selectively at the apical plasma membrane of MV infected cells. M did not colocalize with the glycoproteins at basolateral membranes of polarized cells indicating an independent surface transport mechanism. Analysis of infected cells treated with monensin supported this view. When H and F were retained in the medial Golgi by monensin treatment, M did not accumulate in this cellular compartment. To elucidate the subcellular transport mechanism of the cytosolic M protein, M was expressed in the absence of other viral proteins. Flotation analysis demonstrated that most of the M protein coflotated in infected or in M-transfected cells with cellular membranes. Thus, the M protein possesses the intrinsic ability to bind to lipid membranes. Unexpectedly, plasmid-encoded M protein was rarely found to accumulate at surface membranes. Although cotransport with the viral glycoproteins was not needed, M transport to the plasma membrane required a component only provided in MV infected cells. PMID- 11864738 TI - Identification and molecular characterization of the baculovirus CfMNPV early genes: ie-1, ie-2 and pe38. AB - Three early virus genes, ie-1, ie-2 and pe38, were identified and localized in the XbaI G region (91.2-98.6 m.u.) of the genome of Choristoneura fumiferana nucleopolyhedrovirus (CfMNPV), a baculovirus pathogenic to spruce budworm. Nucleotide sequence analysis indicated that these genes share varied sequence similarity with their homologues in other baculoviruses where they are involved in regulating virus gene expression and DNA replication. Sequence motifs characteristic of DNA binding and transactivation found in other baculovirus regulatory genes were conserved in the CfMNPV genes. Northern analysis demonstrated that all three CfMNPV genes were transcriptionally active in virus infected cells and followed the temporal expression pattern of immediate early baculovirus genes. Primer extension experiments revealed that typical baculovirus early transcription start sites (CAGT) were used for ie-1 and pe38 transcription initiation. Two regions of highly repetitive DNA were found in the odv-e56 to ie 2 and ie-2 and pe38 intergenic regions. These sequences are predicted to function as transcriptional enhancers and viral origins of DNA replication. PMID- 11864739 TI - Both the P and V proteins of the porcine rubulavirus LPMV interact with the NP protein via their respective C-terminal unique parts. AB - In this paper we show that the porcine rubulavirus LPMV phosphoprotein (P) and V protein (V) both interact with the nucleoprotein (NP). There are also indications for an interaction between P and V with L protein. Further analysis of the domains of the P and V which are necessary for interaction with the NP protein demonstrates that the interaction is not mediated from their common part but instead from their unique C-terminal parts, respectively. The common N-terminus of P and V appear to mediate the interaction with L. We also map the regions of NP that are necessary for interaction with P and V, respectively. Both P and V interact with regions of NP, which reside in the N-terminal part but appear not to overlap. PMID- 11864740 TI - Use of pseudotyped retroviral vectors to analyze the receptor-binding pocket of hemagglutinin from a pathogenic avian influenza A virus (H7 subtype). AB - The hemagglutinin (HA) protein of influenza virus binds to terminal sialic acid residues present on cell surface glycoproteins and glycolipids. The specific amino acids involved in this interaction have been identified for a H3 subtype HA from the human non-pathogenic virus, A/Aichi/2/68, by both crystallographic and mutagenesis studies. We were interested to examine the receptor-binding pocket of a H7 subtype protein from the avian pathogenic virus A/FPV/Rostock/34. Accordingly, we made amino acid substitutions at six conserved residues (Y88, T126, H174, E181, L185, and G219), suggested by comparison with the receptor binding pocket of the H3 protein, and analyzed the resulting proteins using pseudotyped retroviral vectors. The use of these vectors enabled us to quantitate both the ability of the mutant HA proteins to bind with receptor-expressing cells, and also to promote virus-cell fusion by measuring vector titer. Using this system, we identified a subset of mutants with impaired receptor-binding activity and a corresponding decrease in titer, but which retained the ability to induce syncytia in low pH cell-cell fusion assays. The most severely affected mutants contained more than one substitution, with the triple mutant Y88F/E181Q/G219K being the most defective. These observations highlight the importance of multiple contact points for the interaction between sialic acid and HA. PMID- 11864741 TI - The insertion of an anti-MHC I ScFv into the N-terminus of an ecotropic MLV glycoprotein does not alter its fusiogenic potential on murine cells. AB - It is known that targeted infection requires the modification of the viral envelope, in order to render it capable of recognizing and specifically binding to a marker protein of the target cell. We have previously described such a recombinant envelope, which is able to extend the tropism of an ecotropic murine leukemia viruses (MLV) envelope to MHC I-expressing human cells. Although, this envelope was very efficient in binding human cells, it yielded very low infection titers. Our attempts to improve these yields by the additional cloning of a variety of spacers in the proximity of the single-chain variable fragment (ScFv) moiety did not significantly influenced human titers, although some alterations on murine titers were observed. To examine whether these low yields represent a decreased fusion capacity of the recombinant envelopes, we performed an assay which allowed the direct comparison between the fusiogenicity of the wild-type (w/t) and the chimeric envelopes. No fusiogenicity of the chimeric envelopes was observed when chimera-expressing cells were co-cultured with human cells. The inability of the chimeras to induce fusion after binding of the ScFv moiety to its ligand may explain, in part, the low infection titers on human cells. However, the several-fold differences observed between the titers of the w/t envelope and the various chimeras on murine cells were not reflected on their fusiogenic potentials, which were all in the same order of magnitude. Our results demonstrate that the binding of the ScFv moiety to its ligand induces no fusion, albeit its insertion into the envelope does not alter the intrinsic fusiogenic ability of the latter. Induction of fusion results from the binding of the envelope to the ecotropic receptor, without being directly proportional to its binding affinity. Chimeras with different infection titers on murine cells yielded similar syncytia counts after their binding to the ecotropic receptor. PMID- 11864742 TI - Nucleotide sequence and construction of an infectious cDNA clone of an EMCV strain isolated from aborted swine fetus. AB - A full-length cDNA clone of an Encephalomyocarditis virus (EMCV) strain (2887A) isolated from aborted swine fetus was constructed and sequenced. Sequence comparison showed more than 99% nucleotide and amino acid sequence identity with two other EMCV strains, EMCV-PV21 and -R. However, the 2887A genomic sequence showed only about 84% nucleotide identity and 96% amino acid identity with EMCV B, -D and -PV2 variants. RNA synthesized by in vitro transcription of this cDNA clone was infectious upon transfection of BHK21 cells, as shown by cytopathic effects and identification by neutralization test, and by propagation of the virus released into the culture media. The transcript RNA led to the production of infectious particles despite the presence of two nongenomic nucleotide residues at the 5' end, the short poly(C) tract (C(10)TCTC(3)TC(10)), the short poly(A) tail (7A), and the presence of six nongenomic nucleotides at the 3' end. The rescued virus was also found to be highly pathogenic for mice by intra peritoneal inoculation producing a fatal disease indistinguishable from that of wild-type virus. An important finding concerning the molecular basis of infectivity was that the in vitro synthesized EMCV RNA transcript is infectious, although it contains a very short poly(A). The availability of the infectious cDNA clone of the reproductive failure strain of EMCV should prove to be useful for studying the molecular basis of the pathogenicity of EMCV in pig. PMID- 11864743 TI - Efficient and tightly regulated expression systems for the human cytomegalovirus major transactivator protein IE2p86 in permissive cells. AB - The 86-kDa IE2 protein (IE2p86) of human cytomegalovirus is a pleiotropic regulatory polypeptide that is essential for activation of viral early promoters and thus, for the entire viral replication cycle. Moreover, this protein modulates cellular gene expression and contributes to the pathogenic features of HCMV. The full spectrum of IE2p86 mediated effects on cellular gene expression has not been defined yet, since efficient expression systems for this protein in HCMV permissive cells are, so far, limited. Here, we report the establishment of two efficient model systems that allow a tightly regulated expression of IE2p86 in various permissive cell types including primary human fibroblasts, primary endothelial cells and U373MG cells. Firstly, we generated a tetracycline regulated U373MG cell line, which expresses high levels of IE2p86 upon tetracycline removal from the culture medium. Secondly, a recombinant baculovirus was constructed, which expresses IE2p86 under the control of the HCMV major immediate early enhancer/promoter upon transduction of various cell types. Importantly, IE2p86 was functional in both systems, since strong transactivation of luciferase promoter constructs could be measured. Furthermore, a cell cycle arrest was detectable after infection of primary human fibroblasts with IE2p86 expressing baculoviruses. Both expression systems represent useful tools to fully define the effects of this pleiotropic transactivator on cellular gene expression and to establish screening systems for novel antiviral drugs targeting this critical immediate early protein of HCMV. PMID- 11864744 TI - Characterization of regions in the GP5 protein of porcine reproductive and respiratory syndrome virus required to induce apoptotic cell death. AB - Expression of the GP5 protein of porcine reproductive and respiratory syndrome virus in mammalian cells using a recombinant vaccinia virus has been shown to induce strong cytotoxicity due to apoptotic death. We have now developed a transient expression system that allows the observation and quantitation of the cell death due to GP5 synthesis, taking advantage of the reduction that this protein induces in the expression of two different co-transfected reporter genes. In this way, we are able to study the regions in GP5 implicated in apoptosis induction. The first 119 aminoacids constitute a region capable of fully inducing apoptosis, aminoacids 90-119 having a fundamental role. On the contrary, the C terminal region is unable by itself of cell death induction and, moreover, is dispensable for this phenotype. We have also observed that induction of apoptosis is independent of cleavage of the N-terminal putative signal sequence in GP5 or N glycosylation of this protein. PMID- 11864745 TI - Nucleotide sequence analysis of the Newcastle disease virus nucleocapsid protein gene and phylogenetic relationships among the Paramyxoviridae. AB - The nucleocapsid (N) protein genes from 24 Newcastle disease virus (NDV) isolates representing various pathotypes with different geographical and chronological origins were cloned and sequenced. The N-terminal region of the N protein to residue 401 was highly conserved among isolates with several conservative substitutions occurring that correlated with phylogenetic relationships. Variability of the N protein was detected in the C-terminal portion similar to what has been reported for other members of the Paramyxovirinae. Amino acids previously identified as invariant or highly conserved in N proteins of other paramyxoviruses were also present in the NDV protein. Phylogenetic analysis of N gene coding sequences among NDV isolates again demonstrated the existence of two major groups. One clade contained viruses that included vaccine and virulent strains isolated in the USA prior to 1970 while a second clade included vaccine and virulent viruses isolated worldwide. Comparison of N protein amino acid sequences among members of the Paramyxoviridae resulted in NDV and avian paramyxovirus 6 separating as a cluster distinct from the Rubulavirus genus. This provides further support for avian paramyxoviruses being considered for their own genus among the Paramyxovirinae. PMID- 11864747 TI - Genetic analysis of canine parvovirus isolates (CPV-2) from dogs in Italy. AB - Genetic and antigenic properties of 62 field isolates of canine parvovirus (CPV 2) collected from 1994 to 2001 in Italy were investigated. Antigenic characterisation was conducted using specific monoclonal antibodies (Mabs). The VP1?VP2 gene was amplified by PCR and characterised with restriction endonucleases to detect the 297 and 265 variant. The VP2 gene of 16 isolates was sequenced and molecular genetic analysis was conducted. The antigenic type prevalent among our isolates is type 2a as well as the 297 variant, which is also prevalent in the rest of Europe. Only the 9.7% of the isolates have the T265P mutation. The VP2 sequences of CPV-2 isolates were very similar to recent Asian isolates. In the threefold spike of CPV-699 a coding change was detected in the 440 residue where threonine was substituted by alanine: the same mutation has been found in two Asian CPV-2 isolates from leopard cats [Virology 278 (2000) 13]. Phylogenetic analysis revealed that the Italian CPV-2 strains followed the same evolution as observed in other countries and they gave no indication of a separate lineage. PMID- 11864746 TI - Expression of a foreign gene by recombinant canine distemper virus recovered from cloned DNAs. AB - A canine distemper virus (CDV) genomic cDNA clone and expression plasmids required to establish a CDV rescue system were generated from a laboratory adapted strain of the Onderstepoort vaccine virus. In addition, a CDV minireplicon was prepared and used in transient expression studies performed to identify optimal virus rescue conditions. Results from the transient expression experiments indicated that minireplicon-encoded reporter gene activity was increased when transfected cell cultures were maintained at 32 rather than 37 degrees C, and when the cellular stress response was induced by heat shock. Applying these findings to rescue of recombinant CDV (rCDV) resulted in efficient recovery of virus after transfected HEp2 or A549 cells were co-cultured with Vero cell monolayers. Nucleotide sequence determination and analysis of restriction site polymorphisms confirmed that rescued virus was rCDV. A rCDV strain also was engineered that contained the luciferase gene inserted between the P and M genes; this virus directed high levels of luciferase expression in infected cells. PMID- 11864748 TI - Specificity analysis of the conserved hexanucleotides for the replication of bamboo mosaic potexvirus RNA. AB - In order to investigate the possible function of the potexviral conserved hexanucleotide sequence (ACc/uUAA) found in the 3' untranslated region of bamboo mosaic potexvirus (BaMV) genomic RNA, each nucleotide in the hexamer motif was substituted. Transcripts derived from wild-type and mutants with a loop deletion or a single-nucleotide substitution were inoculated into protoplasts. The accumulation levels of viral coat protein and RNAs detected from Western and Northern blots of each inoculation were examined after a 48-h incubation. Our data revealed that the nucleotides at positions 4-6 of the hexamer motif cannot be replaced by other nucleotides; the first position of this hexamer is purine specific, and the second position is restricted to pyrimidine. Substitution at the third position has less effect on viral accumulation in protoplasts. In addition to the results reported previously that the E. coli over-expressed BaMV RNA-dependent RNA polymerase could specifically interact with the hexamer motif, the hexanucleotides were thought to serve as a recognition site of viral replicase and the specificity may be derived from the functional groups of each nucleotide of this hexamer motif. PMID- 11864749 TI - Acute hepatic failure in IFN-gamma-deficient BALB/c mice after murine coronavirus infection. AB - We previously showed that an intraperitoneal infection with mouse hepatitis virus (MHV) persists in interferon-gamma (IFN-gamma)-deficient C57BL/6 (B6-GKO) mice and results in subacute fatal peritonitis, which bears a resemblance to feline infectious peritonitis. To examine the role of other host factors in MHV infection in mice, IFN-gamma-deficient mice with a BALB/c background (BALB-GKO) were infected intraperitoneally with MHV and compared with B6-GKO mice. In contrast to B6-GKO mice, BALB-GKO mice died within 1 week due to acute hepatic failure. The viral titer of the liver in BALB-GKO mice was significantly higher than that in B6-GKO mice. All hepatocytes in BALB-GKO mice were necrotic at 5 days post-infection, which was clearly distinct from large but limited lesion in the liver from infected B6-GKO mice. The serum alanine aminotransferase activity of infected BALB-GKO mice were higher than that of B6-GKO mice and was paralleled with the severity of the pathological changes and viral titers in infected mice. Administration of exogenous IFN-gamma to BALB-GKO partially inhibited the acute death. These results indicate that BALB-GKO and B6-GKO mice clearly show different diseases following MHV infection, although wild type counterparts of both mice apparently showed the same clinical course after MHV infection. PMID- 11864750 TI - Human group C rotavirus: completion of the genome sequence and gene coding assignments of a non-cultivatable rotavirus. AB - Genome segments 1 and 2 of human group C rotavirus 'Bristol' strain were sequenced and their gene-protein coding properties assigned. This work completed the genome sequence of a human group C rotavirus (17,910 bp) and allowed the full gene-protein coding assignment of the 11 segments of dsRNA. Gene 1 is 3309 bp in size and contains a single ORF of 3272 nucleotides, encoding a protein of 1090 amino acids in length with a predicted molecular mass of 125 kDa. Comparison of the translated sequence with cognate published mammalian group A, B and C rotavirus sequences showed 45.2, 26.4 and 92.6% identity, respectively. The sequence contains conserved amino acid motifs including the classic RNA-dependent RNA polymerase motif GDD, indicating that segment 1 encodes the group C rotavirus polymerase protein. Gene 2 is 2736 bp in size and contains a single ORF of 2655 nucleotides encoding a protein of 884 amino acids in length with a calculated molecular mass of 102 kDa. Database searches showed highest homology with VP2, the main structural component of the 'core' from group A rotaviruses (46% identity). Alignment of the human group C and A rotavirus VP2 proteins revealed several characteristics common to nucleic acid binding proteins. However, these features were not shared with group B rotavirus VP2. PMID- 11864751 TI - Molecular confirmation of an adenovirus in brushtail possums (Trichosurus vulpecula). AB - Partial genome characterisation of a non-cultivable marsupial adenovirus is described. Adenovirus-like particles were found by electron microscopy (EM) in the intestinal contents of brushtail possums (Trichosurus vulpecula) in New Zealand. Using degenerate PCR primers complementary to the most conserved genome regions of adenoviruses, the complete nucleotide sequence of the penton base gene, and partial nucleotide sequences of the DNA polymerase, hexon, and pVII genes were obtained. Phylogenetic analysis of the penton base gene strongly suggested that the brushtail possum adenovirus (candidate PoAdV-1) belongs to the recently proposed genus Atadenovirus. Sequence analysis of the PCR products amplified from the intestinal contents of brushtail possums originating from different geographical regions of New Zealand identified a single genotype. This is the first report of molecular confirmation of an adenovirus in a marsupial. PMID- 11864752 TI - Unscheduled expression of capsid protein IIIa results in defects in adenovirus major late mRNA and protein expression. AB - Adenovirus gene expression is to a large extent regulated at the level of alternative RNA splicing. For example, in the major late region 1 (L1) unit, a common 5' splice site can be joined to two alternative 3' splice sites, resulting in the formation of the so-called 52,55K (proximal 3' splice site) or the IIIa (distal 3' splice site) mRNAs. Whereas, the 52,55K mRNA is expressed both early and late during infection, the IIIa mRNA is strictly confined to the late phase of the infectious cycle. We have previously shown that IIIa mRNA splicing is subjected to a tight viral control of IIIa 3 splice site usage. In an attempt to determine why adenovirus uses elaborate mechanisms to confine IIIa mRNA production to the late phase of infection, we characterized the phenotype of a recombinant adenovirus expressing the IIIa protein from an inducible tetracycline regulated gene cassette. The results show that expression of the IIIa protein during the early phase of infection results in a significant reduction in late viral protein synthesis and a moderate block to viral DNA replication. Interestingly, unscheduled IIIa protein expression resulted in a perturbation of the accumulation of alternatively spliced L1 mRNAs. Thus, 52,55K mRNA accumulation was inhibited while no effects on endogenous IIIa mRNA expression was detected. PMID- 11864753 TI - Isolation of avian pneumovirus from mallard ducks that is genetically similar to viruses isolated from neighboring commercial turkeys. AB - Our earlier studies demonstrating avian pneumovirus (APV) RNA in wild geese, sparrows, swallows, starlings and mallard ducks suggested that wild birds might be involved in the circulation of APV in the United States. To determine whether turkey virus can be transmitted to the free flying birds, we placed APV-negative mallard ducks next to a turkey farm experiencing a severe APV outbreak and in an area with a large population of waterfowls. The sentinel ducks did not develop clinical APV disease but infectious APV (APV/MN-12) was recovered from choanal swabs after 2 weeks, and anti-APV antibodies detected after 4 weeks. Four APV isolates recovered from the neighboring turkeys that were experiencing an APV outbreak at the same time shared 95-99% nucleotide identity and 97-99% predicted amino acid identity with the duck isolate. In addition experimental infection of turkey poults with APV/MN-12 resulted in detection of viral RNA in nasal turbinates and APV-specific IgG in serum. These results indicate that the APV isolates from turkeys and ducks shared a common source, and the viruses from different avian species can cross-infect. PMID- 11864754 TI - A carboxyl-terminal serine of the bovine papillomavirus E1 protein is phosphorylated in vivo and in vitro. AB - The E1 protein of bovine papillomavirus (BPV) plays several key roles in viral DNA replication. E1 binds the viral origin, unwinds template DNA at the replication fork and recruits cellular replication machinery to the viral DNA. E1 is phosphorylated at multiple sites, and phosphorylation of E1 regulates E1 function and viral DNA replication. A consensus motif for the cellular kinase CK2 was identified at serine 584 near the carboxyl terminus of BPV E1, and found to be highly conserved among papillomavirus E1 proteins. Serine 584 was identified as a substrate of CK1 and CK2 in vitro by mutational and biochemical analysis, and was phosphorylated by a cellular kinase in cultured cells. PMID- 11864757 TI - Ageratum conyzoides L. (Asteraceae). AB - Ageratum conyzoides L., is an annual herb with a long history of traditional medicinal uses in many countries in the world, especially in the tropical and subtropical regions. A wide range of chemical compounds including alkaloids, flavonoids, chromenes, benzofurans and terpenoids have been isolated from this species. Extracts and metabolites from this plant have been found to possess pharmacological and insecticidal activities. The comprehensive account of the chemical constituents and the biological activities are presented in this review such that the potential use of this plant either in pharmaceutics or as an agricultural resource can be evaluated. PMID- 11864758 TI - Antitrypanosomal effect of the aqueous extract of Brassica oleracea. AB - The in vitro antitrypanosomal activity of the aqueous extract of Brassica oleracea, was investigated in Trypanosoma brucei brucei 'Lafia' strain. The extract was found to be effective by immobilizing the trypanosomes within the 3-h incubation period and thereafter rendering them not infective to mice. PMID- 11864759 TI - A new pyrrole alkaloid from seeds of Castanea sativa. AB - A new pyrrole alkaloid, methyl-(5-formyl-1H-pyrrole-2-yl)-4-hydroxybutyrate (1), was isolated from sweet chestnut seeds and its structure elucidated on the basis of data from NMR spectroscopy and by comparison with synthetic analogues. PMID- 11864760 TI - Diuretic, laxative and toxicity studies of Cocculus hirsutus aerial parts. AB - The aqueous extract of the Cocculus hirsutus aerial parts (100 and 200 mg/kg, p.o.) showed significant diuretic activity and laxative effect in rats. The acute toxicity, orally evaluated in mice, was found to be higher than 3000 mg/kg. PMID- 11864761 TI - Organogenesis and terpenoid synthesis in Mentha arvensis. AB - Leaf discs obtained from field grown plants of Mentha arvensis were used to initiate multiple shoots on Murashige and Skoog (MS) medium supplemented with 6 benzylaminopurine (5 mg l(-1)) and naphthaleneacetic acid (0.5 mg l(-1)). Profuse rooting was achieved when the well-grown shoots were cultured on half strength MS medium supplemented with indole-3-acetic acid (2 mg l(-1)). The regenerated plantlets were hardened and successfully transferred to soil and grown to maturity. Tissues at different stages of differentiation were analyzed for their essential oil content and characteristic monoterpene pattern. Tissue culture raised plants show the same essential oil profile as that of the parent plant. However, tissues at early stages of growth show distinct changes in oil composition, such as high levels of pulegone in shoot cultures. PMID- 11864762 TI - Constituents of Gnetum montanum. AB - A new stilbene, gnetifolin M (1), was isolated from the lianas of Gnetum montanum, together with seven known compounds, resveratrol (2), gnetol (3), 4', 5,7-trihydroxy-3'-methoxyflavone, beta-sitosterol, daucosterol, ursolic acid, and tetracosanoic acid. The structure of 1 was determined to be 2-(5'-methoxy-3' hydroxyphenyl)-4-hydroxybenzofuran on the basis of spectroscopic evidence. PMID- 11864763 TI - Antidiarrhoeal activity of Strychnos potatorum seed extract in rats. AB - The antidiarrhoeal activity of the methanol extract of the dried seeds of Strychnos potatorum (MESP) has been evaluated out in rats using different models (castor oil-induced diarrhoea, effects on gastrointestinal motility and on PGE(2) induced gastric enteropooling. MESP (100, 200 and 400 mg/kg, p.o.) significantly (P<0.001) inhibited the frequency of defaecation and reduced the wetness of faecal droppings in castor oil-induced diarrhoea, decreased the propulsion of charcoal meal through the gastrointestinal tract, and also reduced the PGE(2) induced enteropooling. PMID- 11864764 TI - Furanosesquiterpenes from Commiphora sphaerocarpa and related adulterants of true myrrh. AB - A new furanosesquiterpene, (1E)-8,12-epoxygermacra-1,7,10,11-tetraen-6-one (3), was isolated from the resin of Commiphora sphaerocarpa together with the known compounds curzerenone (1), furanodienone (2), (1E)-3-methoxy-8,12-epoxygermacra 1,7,10,11-tetraen-6-one (4), (1(10)E,2R*,4R*)-2-methoxy-8,12-epoxygermacra- 1(10),7,11-trien-6-one (5), and dihydropyrocurzerenone (6). Hydrodistillates of the resins of C. sphaerocarpa, C. holtziana, C. kataf and C. myrrha were analysed. The identifications were aided by NMR, GC and GC-MS. PMID- 11864766 TI - Lotusine G: a new cyclopeptide alkaloid from Zizyphus lotus. AB - In addition to six cyclopeptide alkaloids lotusines A-F, a new compound lotusine G, was isolated from the root bark of Zizyphus lotus by centrifugal partition chromatography. PMID- 11864765 TI - Inhibitory action of kaurenoic acid from Viguiera robusta (Asteraceae) on phenylephrine-induced rat carotid contraction. AB - ent-kaurenoic acid (KA) isolated from Viguiera robusta was tested for activity on vascular smooth muscle contactility. Cumulative concentration-response curves were obtained for a stepwise increase (10(-10)-10(-05) mol/l) in the concentration of phenylephrine (Phe), a selective alpha(1)-adrenergic agonist. The effects in the presence of KA (0.2, 2.0 and 20.0 microg/ml), and 90 min after the removal of KA from the medium bath were compared to controls. At 20.0 microg/ml, KA inhibited the in vitro contractility of rat carotid artery elicited by Phe, but had no effect at lower concentrations (0.2 and 2.0 microg/ml). The effect elicited by KA was reversible after 90 minutes. PMID- 11864767 TI - Medicinal plants popularly used in the Brazilian Tropical Atlantic Forest. AB - A survey of medicinal plants used by rural and urban inhabitants of the three cities of the Tropical Atlantic Forest, Region of Vale do Ribeira, State of Sao Paulo, Brazil was performed by means of 200 interviews with medicinal plant users and extractors and, traditional healers. One hundred fourteen herbal remedies were recorded and the following information reported: Latin, vernacular and English names, plant part used, forms of preparation and application of the herbal remedies, medicinal or food uses, areas of plant collection, economic importance (when available) and other data. PMID- 11864768 TI - Antimicrobial activity and cytotoxicity of Aerva lanata. AB - Aerva lanata whole plant showed interesting antimicrobial activities (ethyl acetate and methanol extracts) and significant cytotoxic properties (petroleum ether, ethyl acetate and methanol extracts). PMID- 11864769 TI - A novel acylated flavonol glycoside from Morina nepalensis var. alba. AB - A novel acylated flavonol glycoside, quercetin 3-O-[2' "-O-(E)-caffeoyl]-alpha-L arabinopyranosyl-(1-->6)-beta-D-galactopyranoside (1), was isolated from whole plant of Morina nepalensis var. alba. Its structure was determined on the basis of chemical and spectroscopic methods. PMID- 11864770 TI - A new eudesmane derivative from Onopordon ambiguum. AB - Reinvestigation of Onopordon ambiguum extract afforded a new eudesmane derivative 1, in addition to the known elemanoid derivative 2. The structures of both compounds were determined by spectroscopic techniques, including 1H-, 13C-NMR, DEPT, 1H-1H and 1H-13C COSY analysis. PMID- 11864772 TI - An analysis of travel costs on transport of load and nest building in golden hamster. AB - We investigated the effects of travel costs on transporting nest material and nest-building activity in golden hamsters. Nest-deprived animals were submitted to run alleys 30, 90 and 180 cm long to access a source containing paper strips as nest material (Experiment 1) or were submitted to the same travel costs in 24 h experimental sessions (Experiment 2). We noted that increased travel costs were related to a decreased number of trips to the source, larger amounts (cm(2)) of nest material transported per trip (although total loads also decreased in longer alleys), longer intervals between trips, and increased time spent at the source and in nest building activity. Foraging efficiency (i.e. size of load divided by the time spent at the source) decreased as a function of travel costs, and animals transported their loads in two fundamental ways: in 30-cm alleys, they simply used their mouth to pull the paper strips, but in 90- or 180-cm alleys they transported the loads in their cheek pouches. The animals were faster when returning to the home-cage and their running speed (cm/s) increased as a function of the length of the alley, showing that animals are under different environmental pressures when searching for resources and subsequently running back with the load to the nest. Both male and female subjects were sensitive to travel costs, but males engaged in nest building activity more promptly and exhibited higher mean performances in most measures. We conclude that nest material is a good reinforcer, and our major results are in accordance with the predictions of microeconomic and optimal foraging theories. PMID- 11864771 TI - Acquired equivalence of flavour cues with a common antecedent in rats. AB - Equivalence of flavour cues, each of which had been paired with a common antecedent, was demonstrated with rats in a three-stage design. In the first stage, a group of thirsty rats were given each of two target flavour cues after a common antecedent flavour (Xright arrowA and Xright arrowB), while a second group of rats were given A and B after differential antecedent flavours (Xright arrowA and Yright arrowB). Another group of rats was allowed to drink A and B after familiar tap water. In the second stage, aversion to A was established by a lithium chloride injection after drinking A. The acquired equivalence effect was verified in the third stage by strong aversion to B in the group trained with the common antecedent compared with the remaining two groups. The representation mediation, rather than response-mediation, hypothesis seems to fit the backward acquired equivalence effect obtained here. PMID- 11864773 TI - Rule-governed versus contingency-governed behavior in a self-control task: effects of changes in contingencies. AB - Rule-governed behavior is typically acquired faster than contingency-governed behavior but is less sensitive than contingency-governed behavior to unverbalized contingency changes. The present study investigated these relationships in a computer task frequently used to study human self-control. Instructions for one group of participants contained a hint about how to maximize long-term reinforcement; the other group performed the task without the hint. Participants given the hint came closer to maximizing reinforcement in the long term, but their behavior was less sensitive to an unsignaled contingency change than that of those not given the hint. The study shows that, like other complex behaviors, self-control may be contingency-governed or rule-governed. PMID- 11864774 TI - Maternal-care behavior and life-history traits in house mice (Mus domesticus) artificially selected for high voluntary wheel-running activity. AB - To test the hypothesis that selective breeding for high voluntary wheel running negatively affects maternal performance in house mice, we observed maternal behavior and compared litter size and mass, in replicate lines of selected (N=4) and control (N=4) mice from generations 20 and 21 of an artificial selection experiment. At generation 21, selected-line females ran 2.8-times more revolutions per day than females from random-bred control lines, when tested at approximately 6 weeks of age as part of the normal selection protocol. After giving birth, dams from selected and control lines exhibited similar frequencies of maternal behaviors and also spent similar amounts of time in general locomotor activity at litter ages of both 9 and 16 days. Dams from selected lines also performed equally well as controls in repeated pup-retrieval trials. At first parturition, selected-line dams averaged 2.4 g smaller in body mass as compared with dams from the control lines; however, neither litter size nor litter mass at birth (generation 20) or at weaning (generation 21) differed significantly between selected and control lines. We conclude that, at least under the husbandry conditions employed, maternal behavior and reproductive output at first parturition are genetically independent of wheel-running behavior. PMID- 11864776 TI - Editorial. PMID- 11864775 TI - Instrumental responding remains under the control of the consequent outcome after extended training. AB - The effect of extensive training on the contribution of response--outcome learning to instrumental performance in rats was re-examined in two experiments using a transfer test. In each experiment, two discriminative stimuli were established as signals for different response--outcome combinations (e.g. light: nose poke-pellets and noise: handle pull-sucrose). Then, two different responses (lever press and chain pull) were concurrently trained with those outcomes. In Experiment 1, these responses underwent extensive training, each with a different outcome (e.g. lever press-pellets and chain pull-sucrose). In Experiment 2, these responses were trained moderately with one outcome (e.g. lever press-pellets and chain pull-sucrose) and extensively with a different outcome (e.g. lever press sucrose and chain pull-pellets). Finally, transfer tests were conducted in which the discriminative stimuli, noise and light, were tested periodically with the lever and chain. In Experiment 1, the stimuli consistently and preferentially elevated performance of the response trained with the same outcome relative to that trained with a different outcome. In Experiment 2, the stimuli elevated both responses nondifferentially. However, a drive manipulation (thirst) designed to increase the value of sucrose relative to pellets revealed a significant preference for the response that had been extensively trained with sucrose. Overall, the results of these experiments confirm previous findings that instrumental behaviors do not become increasingly independent of their consequent outcomes with extended training. However, the transfer results of Experiment 2 highlight a potential limitation of the transfer test for assessing variations in the strength of R--O associations. PMID- 11864777 TI - The role of advanced oxidation protein products in regulation of dendritic cell function. AB - The basis for this study was the "injury hypothesis," which holds that release of micro-environmental constituents, such as reactive oxygen species and oxidants, acts as a signal, and potential activator, of dendritic cell (DC)-mediated antigen presentation. Following this oxidative stress, dityrosine containing cross-linked proteins, advanced oxidation protein products (AOPP), are known to be generated, and we proposed that they may serve as moieties that mediate such signals. Therefore, the effect of AOPP on DCs has been examined in vitro. There were no AOPP-induced changes in DC phenotype as judged by expression of typical surface costimulatory molecules. However, at higher cell concentrations AOPP treated DCs were more potent inducers in an oxidative mitogenesis assay than controls. Thus, AOPP may act like superantigens, allowing for bypass of upregulation of costimulation, and, either alone or in synergy with oxidants themselves, serving as amplifiers of DC function. PMID- 11864778 TI - Flavonoids increase the intracellular glutathione level by transactivation of the gamma-glutamylcysteine synthetase catalytical subunit promoter. AB - Fruits and vegetables protect against cancer by so far not well-characterized mechanisms. One likely explanation for this effect is that dietary plants contain substances able to control basic cellular processes such as the endogenous defense against oxidative stress. Oxidative stress is pivotal in many pathological processes and reduced oxidative stress is implicated in prevention of disease. Our results demonstrate that extract from onion and various flavonoids induce the cellular antioxidant system. Onion extract and quercetin were able to increase the intracellular concentration of glutathione by approximately 50%. Using a reporter construct where reporter expression is driven by the gamma-glutamylcysteine synthetase (GCS) heavy subunit (GCS(h)) promoter we show that onion extract, quercetin, kaempferol, and apigenin increased reporter gene activity, while a fourth flavonoid, myricetin and sugar conjugates of quercetin were unable to increase reporter expression. Quercetin was also able to induce a distal part of the GCS(h) promoter containing only two antioxidant response/electrophile-response elements (ARE/EpRE). Our data strongly suggest that flavonoids are important in the regulation of the intracellular glutathione levels. This effect may be exerted in part through GCS gene regulation, and may also contribute to the disease-preventing effect of fruits and vegetables. PMID- 11864780 TI - Hydrogen peroxide regulation of bovine endothelin-converting enzyme-1. AB - Vascular injury leads to the production of reactive oxygen species (ROS), but the mechanisms by which ROS contribute to vascular pathology are not completely understood. We hypothesized that ROS increase endothelin converting enzyme (ECE 1) expression. We found that glucose oxidase (GO) increases ECE-1 mRNA, protein, and activity in bovine aortic endothelial cells. Catalase abolishes this effect. Glucose oxidase treatment of endothelial cells transactivates the ECE-1 promoter. The ECE-1 promoter element that mediates this response to GO is located between 444 and -216 bp. This region contains a STAT response element, and GO activates STAT-3 binding to this STAT response element. Our data suggest that STAT3 mediates hydrogen peroxide induction of ECE-1 expression. PMID- 11864779 TI - Imidazoline drugs stabilize lysosomes and inhibit oxidative cytotoxicity in astrocytes. AB - Oxidative stress is a primary pathogenesis in the brain, which is particularly vulnerable to oxidative stress. Maintenance of astrocyte functions under oxidative stress is essential to prevent neuronal injuries and to recover neuronal functions in various pathologic conditions. Imidazoline drugs have affinities for imidazoline receptors, which are highly distributed in the brain, and have been shown to be neuroprotective. This study presented the protective effects of several imidazoline drugs against oxidative cytotoxicity, in primary cultures of astrocytes. Imidazoline drugs, such as idazoxan, guanabenz, guanfacine, BU224, and RS-45041-190, showed protective effects against naphthazarin-induced oxidative cytotoxicity, as evidenced by LDH release and Hoechst 33342/propidium iodide staining. The imidazoline drugs stabilized lysosomes and inhibited naphthazarin-induced lysosomal destabilization, as evidenced by acridine orange relocation. Guanabenz inhibited, the leakage of lysosomal cathepsin D to cytosol, the decreased mitochondrial potential, and the release of mitochondrial cytochrome c, which were induced by naphthazarin. The lysosomal destabilization by oxidative stress and other apoptotic signals and subsequent cathepsin D leakage to the cytosol can induce apoptotic changes of mitochondria and eventually cell death. Therefore, lysosomal stabilization by imidazoline drugs may be ascribed to their protective effects against oxidative cytotoxicity. PMID- 11864782 TI - Control of oxygen free radical formation from mitochondrial complex I: roles for protein kinase A and pyruvate dehydrogenase kinase. AB - Human NADH CoQ oxidoreductase is composed of a total of 43 subunits and has been demonstrated to be a major site for the production of superoxide by mitochondria. Incubation of rat heart mitochondria with ATP resulted in the phosphorylation of two mitochondrial membrane proteins, one with a M(r) of 6 kDa consistent with the NDUFA1 (MWFE), and one at 18kDa consistent with either NDUFS4 (AQDQ) or NDUFB7 (B18). Phosphorylation of both subunits was enhanced by cAMP derivatives and protein kinase A (PKA) and was inhibited by PKA inhibitors (PKAi). When mitochondrial membranes were incubated with pyruvate dehydrogenase kinase, phosphorylation of an 18kDa protein but not a 6kDa protein was observed. NADH cytochrome c reductase activity was decreased and superoxide production rates with NADH as substrate were increased. On the other hand, with protein kinase A driven phosphorylation, NADH cytochrome c reductase was increased and superoxide production decreased. Overall there was a 4-fold variation in electron transport rates observable at the extremes of these phosphorylation events. This suggests that electron flow through complex I and the production of oxygen free radicals can be regulated by phosphorylation events. In light of these observations we discuss a potential model for the dual regulation of complex I and the production of oxygen free radicals by both PKA and PDH kinase. PMID- 11864781 TI - Supplementation with antioxidant vitamins prevents oxidative modification of DNA in lymphocytes of HIV-infected patients. AB - There is evidence suggesting that patients infected with human immunodeficiency virus (HIV) are under chronic oxidative stress. In the present study, the level of oxidatively modified bases in lymphocyte DNA and some other parameters of oxidative stress were measured in HIV-infected patients (n = 30), as well as in control groups (10 healthy volunteers and 15 HIV-seronegative injected drug users). Additional experiments were conducted using lymphocyte DNA samples from asymptomatic seropositive, HIV-infected patients who were supplemented with antioxidant vitamins A, C, and E or received placebo. Significant increases in the amount of the modified DNA bases were observed in HIV-infected patients when compared with the control group. The concentration of thiobarbituric acid reactive substances (TBARS) was higher and activities of antioxidant enzymes (superoxide dismutase and catalase) were lower in the group of HIV-infected patients in comparison to the control group. Vitamin supplementation resulted in the significant decrease in the levels of all modified DNA bases when compared to the patients who received placebo. The reduction of TBARS and the restoration of the activity of the enzymes were also observed. Our data suggest that people infected with HIV can benefit from treatment with antioxidant vitamins. PMID- 11864783 TI - Reactive oxygen species modulate Zn(2+)-induced apoptosis in cancer cells. AB - Some recent evidence has suggested a protective role of zinc against cancer. The mechanism by which zinc exerts this action has not been defined and, in particular, it has not been clarified whether zinc may directly act on cancer cells and the molecular mechanisms involved in this effect. In this study, we examined the in vitro effect of zinc on the apoptosis of mouse TS/A mammary adenocarcinoma cells, studying the zinc-dependent modulation of the intracellular levels of reactive oxygen species (ROS) and of p53 and Fas/Fas ligand pathways. We showed that zinc concentrations ranging from 33.7 to 75 muM Zn(2+) induced apoptosis in mammary cancer cells. The apoptosis was associated with an increased production of intracellular ROS, and of p53 and Fas/Fas ligand mRNA and protein. Zn(2+) induced a faint metallothionein response in TS/A cells in comparison with mouse lymphocytes. The treatment of tumor cells with the antioxidant N acetylcysteine was able to prevent Zn(2+)-induced apoptosis, as well as the increase of p53 and Fas ligand protein induced by zinc. The data demonstrate that zinc exerts a direct action on mammary cancer cells inducing ROS-mediated apoptosis and that the effect may be mediated by the ROS-dependent induction of p53 and Fas/Fas ligand. PMID- 11864784 TI - Overestimation of NADH-driven vascular oxidase activity due to lucigenin artifacts. AB - Several limitations have recently been described for lucigenin, a probe frequently used to assess the activity of vascular NAD(P)H oxidase, a major superoxide source. The preferential reducing substrate of such oxidase remains unclear. We assessed whether lucigenin artifacts could affect detection of NAD(P)H oxidase activity. Initial chemiluminescence assays were performed with vascular rings or homogenates at 5, 50, or 250 microM concentrations. Results showed preferential signals with NADPH (vs. NADH) with 5 and 50 microM lucigenin, which were blocked by diphenylene iodonium (DPI), superoxide dismutase (SOD), or its cell-permeable mimetic MnTBAP. With 250 microM lucigenin, the relative signal with NADH became larger than with NADPH, and was poorly inhibited by all three antagonists above. All SOD/DPI-resistant signals were effectively blocked by the electron acceptor nitrobluetetrazolium. Spin trapping with DMPO showed an approximate doubling of DMPO-OH radical adduct signal upon addition of 5 microM lucigenin to homogenates incubated with either NADPH or NADH. With 50 or 250 microM lucigenin, much larger increases were observed with NADH, as opposed to NADPH. Furthermore, oxygen consumption measurements showed analogous results. In summary, our data suggest that: (i) Lucigenin redox-cycling is detectable in vascular tissue even at 5 microM concentrations, while at 250 microM redox cycling becomes predominant and is markedly increased when NADH is the assayed substrate; and (ii) With 250 microM lucigenin, preferentially with NADH, signals are further overestimated by direct, oxidase-dependent, superoxide-independent two-electron transfer. Therefore, previous reports of preferential NADH affinity of the vascular oxidase may have been due to these artifacts. PMID- 11864785 TI - Ebselen suppresses late airway responses and airway inflammation in guinea pigs. AB - Although ebselen, a seleno-organic compound, inhibits inflammation in various animal models, its efficacy as an anti-asthma drug remains to be clarified. In this study, we investigated the inhibitory effect of ebselen on a guinea pig asthma model. Ebselen was orally administered at dosages of 1-20 mg/kg 2 h before an ovalbumin (OA) challenge, and then airway responses, airway inflammation, the generation of superoxide, H(2)O(2), and nitrotyrosine, and the induction of inducible nitric oxide synthase (iNOS) were evaluated. Sensitized animals challenged with OA aerosol showed dual airflow limitations, i.e., immediate and late airway responses (IAR and LAR). Ebselen significantly inhibited LAR at dosages greater than 10 mg/kg, but did not inhibit IAR at any dosage. Bronchoalveolar lavage (BAL) examination showed that airway inflammation was significantly suppressed by ebselen at 10 mg/kg. The generation of superoxide and H(2)O(2) occurred on endothelial cells of LAR bronchi, and was inhibited by 10 mg/kg of ebselen. Superoxide generation was inhibited by diphenyleneiodonium chloride (DPI), a NAD(P)H oxidase inhibitor, but not by allopurinol, a xanthine oxidase inhibitor. Immunoreactivities for iNOS and nitrotyrosine were also observed on endothelial cells of LAR bronchi and were abolished in ebselen treated animals. The present findings suggest that ebselen can be applied as a new therapeutic agent for asthma. The possible mechanisms by which ebselen inhibits LAR likely involve suppression of oxidant formation and iNOS induction in endothelial cells. PMID- 11864786 TI - Metal-independent production of hydroxyl radicals by halogenated quinones and hydrogen peroxide: an ESR spin trapping study. AB - The metal-independent production of hydroxyl radicals (*OH) from H(2)O(2) and tetrachloro-1,4-benzoquinone (TCBQ), a carcinogenic metabolite of the widely used wood-preservative pentachlorophenol, was studied by electron spin resonance methods. When incubated with the spin trapping agent 5,5-dimethyl-1-pyrroline N oxide (DMPO), TCBQ and H(2)O(2) produced the DMPO/*OH adduct. The formation of DMPO/*OH was markedly inhibited by the *OH scavenging agents dimethyl sulfoxide (DMSO), ethanol, formate, and azide, with the concomitant formation of the characteristic DMPO spin trapping adducts with *CH(3), *CH(CH(3))OH, *COO(-), and *N(3), respectively. The formation of DMPO/*OH and DMPO/*CH(3) from TCBQ and H(2)O(2) in the absence and presence, respectively, of DMSO was inhibited by the trihydroxamate compound desferrioxamine, accompanied by the formation of the desferrioxamine-nitroxide radical. In contrast, DMPO/*OH and DMPO/*CH(3) formation from TCBQ and H(2)O(2) was not affected by the nonhydroxamate iron chelators bathophenanthroline disulfonate, ferrozine, and ferene, as well as the copper-specific chelator bathocuproine disulfonate. A comparative study with ferrous iron and H(2)O(2), the classic Fenton system, strongly supports our conclusion that *OH is produced by TCBQ and H(2)O(2) through a metal-independent mechanism. Metal-independent production of *OH from H(2)O(2) was also observed with several other halogenated quinones. PMID- 11864787 TI - UV-A induces persistent genomic instability in human keratinocytes through an oxidative stress mechanism. AB - Ultraviolet-A (UV-A, 320 to 400 nm) radiation comprises 95% of the solar ultraviolet radiation (UVR) reaching the earth's surface. It has been associated experimentally and epidemiologically with malignant melanoma. In this study we investigated whether UV-A radiation can induce a persistent, heritable hypermutability in mammalian cells similar to that observed following ionising radiation (IR). Using the immortalized human skin keratinocyte cell line HaCaT we found that UV-A radiation does lead to a continuing reduction in plating efficiency, an increased "spontaneous" mutant fraction, and an increase in micronucleus formation up to 21 d after initial exposure. Reversal of these effects using catalase may indicate a role for hydrogen peroxide in this phenomenon. These results add to the significance of UV-A radiation as a risk factor in skin carcinogenesis. PMID- 11864788 TI - Role of inducible nitrogen oxide synthase in benzene-induced oxidative DNA damage in the bone marrow of mice. AB - We investigated the interaction of BZ and lipolysaccharide (LPS), a well-known inflammation-promoting agent, in wild-type and inducible nitrogen oxide synthase (iNOS) knockout mice. BZ generated DNA strand breaks (SB) in the liver of both wild-type and iNOS-deficient mice. In the bone marrow (BM) BZ and LPS generated SB only in wild-type mice. The effects were additive, suggesting that both a redox cycling and an iNOS-dependent pathway may be involved. Formamidopyrimidine DNA glycosylase sensitive sites were elevated by BZ in the BM in both types of mice, whereas endonuclease III sensitive sites were not affected by any treatment. Since BZ is associated with leukemia in humans, it suggests that oxidative DNA base damage rather than SB may be important in the development of leukemia. PMID- 11864790 TI - Predictors of attrition in a longitudinal cognitive aging study: the Maastricht Aging Study (MAAS). AB - A large sample of older participants of the Maastricht Aging Study (MAAS) were compared to drop-outs at the 3-year follow-up with respect to socio-demographic, health, and cognitive characteristics. In addition, the impact of selective drop out on measures of cognitive change was examined. To this end, hypothetical scores were estimated for drop-outs by using single and multiple imputation methods. Of the initial sample of 539 subjects, aged 49 years and older at baseline, 116 (22 %) did not return for the follow-up (n = 32 had died, n = 84 refused participation). Drop-outs who refused to participate in the follow-up were more often women, had lower educational levels, and had lower baseline scores on neurocognitive tests. Follow-up drop-outs who had died were more often men, older, and had a poorer performance on cognitive tests than the follow-up participants. Although follow-up participants and drop-outs differed in terms of socio-demographic and cognitive characteristics, attrition appeared to have little effect on the estimates of cognitive change. PMID- 11864791 TI - Psychological attributes and changes in disability among low-functioning older persons: does attrition affect the outcomes? AB - The impact of attrition was studied in a two-wave panel study on the associations between psychological attributes (mastery, self-efficacy expectancies) and disability increase among low-functioning older persons (n = 753, 76.4% participated at the follow-up 2 years later). The results showed that: (1) attrition at follow-up was clearly associated with age and level of disability at baseline. Older persons and those with higher levels of disability did not participate in the follow-up; and (2) the strength of the association between the selected psychological attributes and the increase in disability did not substantially alter when (a) missing follow-up scores of disability were replaced by several values (e.g., lower, equal or higher compared to those who actually participated at the follow-up), or (b) associations were computed excluding those participants at follow-up with highest increase in disability. We conclude that attrition may affect the descriptive outcomes of aging studies, particularly when such studies focus on health and function, but that attrition not always seems to be a serious problem when associations between variables are the focus of study. PMID- 11864792 TI - Long-term follow-up of participants in clinical studies. AB - We evaluated a multistep protocol to locate and ascertain data from 708 subjects who participated in clinical studies at our medical center up to 27 years ago. The protocol involved computerized and manual chart audits, introductory letters, internet searches, death registries, and a commercial person locator. We were able to ascertain vital status on 596 (84%) of study participants. Moreover, of the 475 persons known to be alive, 426 (90%) returned questionnaires and 275 (58%) completed physical examination and laboratory studies in our Clinical Research Center. The relatively modest cost of the protocol ($53,426) suggests that it is a reasonable strategy to test hypotheses involving long-term outcomes among well-characterized cohorts. The fact that the protocol can be easily implemented in most sites should encourage investigators to use these strategies to conduct similar studies. PMID- 11864793 TI - Urine testing to monitor adherence to TB preventive therapy. AB - This study examined the validity of the Arkansas urine test. One hundred ninety four adolescents submitted an unannounced urine specimen monthly (for 6 to 8 months). Duplicate specimens were blindly tested with high agreement (kappa >90%). Sensitivity and specificity were estimated. In 68% of test runs, adolescents recalled taking INH within 24 hr of specimen collection. For recall intervals of 24, 48, and 72 hr, sensitivity was 87, 85, and 83%, respectively. Females were less likely to test positive when INH was taken within the previous 24 hr (sensitivity 84 versus 92% males). Specificity was 57, 91, and 95% at 24, 48, and 72 hr, respectively. The Arkansas urine test was practical to use, and results correlated well with self-reported adherence to INH for treatment of latent tuberculosis infection (LTBI), over several months of follow-up. The test may be useful as part of an adherence-monitoring program when used in conjunction with self-reported measures. PMID- 11864794 TI - Measurement of fatigue. determining minimally important clinical differences. AB - The purpose was to determine the minimally important clinical difference (MICD) in fatigue as measured by the Profile of Mood States, Schwartz Cancer Fatigue Scale (SCFS), General Fatigue Scale, and a 10-point single-item fatigue measure. The MICD is the smallest amount of change in a symptom (e.g., fatigue) measure that signifies an important change in that symptom. Subjects rated the degree of change in their fatigue over 2 days on a Global Rating Scale. 103 patients were enrolled on this multisite prospective repeated measures design. MICD was determined following established procedures at two time points. Statistically significant changes were observed for moderate and large changes in fatigue, but not for small changes. The scales were sensitive to increases in fatigue over time. The MICD, presented as mean change, for each scale and per item on each scale is: POMS = 5.6, per item = 1.1, SCFS = 5.0, per item = 0.8, GFS = 9.7, per item = 1.0, and the single item measure of fatigue was 2.4 points. This information may be useful in interpreting scale scores and planning studies using these measures. PMID- 11864795 TI - Coaching patients with coronary heart disease to achieve the target cholesterol: a method to bridge the gap between evidence-based medicine and the "real world"- randomized controlled trial. AB - Community studies have demonstrated suboptimal achievement of lipid targets in the management of patients with coronary heart disease (CHD). An effective strategy is required for the application of evidence-based prevention therapy for CHD. The objective of this study was to test coaching as a technique to assist patients in achieving the target cholesterol level of <4.5 mmol/L. Patients with established CHD (n = 245) underwent a stratified randomization by cardiac procedure (coronary artery bypass graft surgery or percutaneous coronary intervention) to receive either the coaching intervention (n = 121) or usual medical care (n = 124). The primary outcome measure was fasting serum total cholesterol (TC), serum triglyceride (TG), high-density lipoprotein cholesterol (HDL-C), and calculated low-density lipoprotein cholesterol (LDL-C) level, measured at 6 months post-randomization. At 6 months, the serum TC and LDL-C levels were significantly lower in the coaching intervention group (n = 107) than the usual care group (n = 112): mean TC (95%CI) 5.00 (4.82-5.17) mmol/L versus 5.54 (5.36-5.72) mmol/L (P <.0001); mean LDL-C (95%CI) 3.11 (2.94-3.29) mmol/L versus 3.57 (3.39-3.75) mmol/L (P <.0004), respectively. Coaching had no impact on TG or on HDL-C levels. Multivariate analysis showed that being coached (P <.001) had an effect of equal magnitude to being prescribed lipid-lowering drug therapy (P <.001). The effectiveness of the coaching intervention is best explained by both adherence to drug therapy and to dietary advice given. Coaching may be an appropriate method to reduce the treatment gap in applying evidence based medicine to the "real world." PMID- 11864796 TI - Increasing incidence of serious hypoglycemia in insulin users. AB - Trials provide most of the data on the incidence of hypoglycemia in patients with diabetes. However, trial protocols may not reflect changes in diabetes management that occur over time. Using a retrospective cohort design, we calculated the incidence of hypoglycemia in 1,113 patients 20 to 64 years of age who used insulin monotherapy from 1993 through 1997 at Harvard Pilgrim Health Care, a health maintenance organization (HMO) in New England. Episodes included emergency department visits or hospital admissions for hypoglycemia identified through insurance claims and documented in patients' medical records. The incidence of hypoglycemia was 5.07 episodes per 100 person-years (95% CI, 4.14 to 6.00); 203 confirmed episodes occurred in 129 unique patients during 4,005 person-years of follow-up. The incidence increased by approximately 24% per year (rate ratio = 1.24; 95% CI, 1.11 to 1.38) from 1993 through 1997, perhaps reflecting physicians' and patients' efforts to intensify glucose management. PMID- 11864797 TI - Lost income and work limitations in persons with chronic respiratory disorders. AB - Chronic respiratory diseases are prevalent, disabling, and rank fifth in terms of cost of illness among major disease groups; however, the degree of work loss and lost income is largely unknown. Lost wages in adults with chronic respiratory conditions were calculated from the Survey of Income and Program Participation (SIPP). The National Health Interview Survey (NHIS) was used to estimate work loss and limitations in specific chronic respiratory conditions, including allergic rhinitis, asthma, chronic bronchitis, and emphysema. The SIPP indicated that adults with chronic respiratory conditions average $3,143 annual earnings loss ($5,272 for those between the ages of 25 and 64 and $1,267 for those over age 65). The NHIS indicated that 19.9% of persons with chronic respiratory conditions report being unable to work. Comparisons across respiratory diseases are made. PMID- 11864798 TI - The aging and dying processes and the health of older adults. AB - It is difficult to distinguish changes in health due to aging from those related to dying, because the two processes are highly related. Some potentially treatable conditions may mistakenly be dismissed as due to old age. The goal of this article was to examine the relationships of aging and of dying to changes in 10 health-related variables: self-rated health, depression, ADLs, IADLs, minimental state examination, body mass index, blocks walked per week, bed days, hospitalization, and walking speed (all coded so that higher values were better). We used longitudinal data from the Cardiovascular Health Study to estimate the changes in the variables associated with 5 years of aging and also in the 5 years before death, controlling for years from death and for age, respectively. All 10 health variables declined as death approached, and most of them also declined with age. The "effect" of the dying process was usually significantly larger than the effect of aging. Large declines in these health measures are probably not due to aging, and should be taken seriously by patients and their providers. PMID- 11864799 TI - Attitudinal factors among adult prescription recipients associated with choice of where to be vaccinated. AB - The objective of this article was to evaluate adult prescription recipients' choices among vaccine providers. The study setting was a cluster sample from 24 community pharmacies based on prescription records that suggested need for pneumococcal and influenza vaccines. Vaccination status, choice of vaccine provider, and opinions regarding vaccine providers were retrospectively surveyed by mail in spring 1999. Overall, 52% of survey recipients responded; 89% of respondents reported returning to the same type of vaccine provider in consecutive years. Two key factors affected choice of provider: convenience and provider experience. Convenience was a stronger factor for people younger than 65 taking chronic medications and those not vaccinated in the previous year. Most adult recipients of influenza vaccine returned to sites where they were vaccinated the previous year. Convenience was a major factor in vaccination decisions of adult prescription recipients. PMID- 11864800 TI - What is a clinically meaningful change on the Functional Assessment of Cancer Therapy-Lung (FACT-L) Questionnaire? Results from Eastern Cooperative Oncology Group (ECOG) Study 5592. AB - To assess the impact of disease and treatment on patients with advanced non-small cell lung cancer (NSCLC), we set out to determine a clinically meaningful change (CMC) on the Lung Cancer Subscale (LCS) and the Trial Outcome Index (TOI) of the Functional Assessment of Cancer Therapy-Lung (FACT-L) questionnaire. We used data from Eastern Cooperative Oncology Group study 5592 (E5592), a randomized trial comparing three chemotherapeutic regimens in 599 advanced NSCLC patients. Patients completed the FACT-L at baseline (pretreatment), 6 weeks, 12 weeks, and 6 months. Comparing across baseline performance status (0 vs. 1), prior weight loss (<5% vs. > or = 5%), and primary disease symptoms (< or = 1 vs. >1), LCS and TOI score differences ranged from 2.4 to 3.6 and 6.5 to 9.2, respectively (all Ps <.001). Mean improvement in LCS score from baseline to 12 weeks was 2.4 points in patients who had responded to treatment versus 0.0 points in patients who had progressive disease. Twelve-week LCS change scores for patients progressing early were 3.1 points worse than those of patients progressing later (mean = -1.2 vs.1.9, respectively). Similarly, the average TOI change score from baseline to 12 weeks was -6.1 for patients who had progressive disease versus -0.8 points for patients who had responded to treatment. Twelve-week TOI change scores for patients progressing early (mean = -8.1) were 5.7 points worse than those of patients progressing later (mean = -8.1 vs. -2.4, respectively). Analyses assuming nonrandom missing data resulted in slightly larger differences. Clinically relevant change scores were estimated as two to three points for the LCS and five to seven points for the TOI, setting upper limits for minimal CMCs. These values were comparable to suggested distribution-based criteria of a minimally important difference. These results support use of a two to three point change in the LCS and five to six point change on the TOI of the FACT-L as a CMC, and offer practical direction for inclusion of important patient-based endpoints in lung cancer clinical trials. PMID- 11864801 TI - The "Adequacy of Prenatal Care Utilization" (APNCU) index to study low birth weight: is the index biased? AB - A recent, nationwide study of 54 million births reported increasing trends toward more prenatal resource utilization from 1981 to 1995, when other indicators have shown worsening trends in birth outcomes. The Adequacy of Prenatal Care Utilization (APNCU) Index was used to measure resource utilization, but the Index appears to be biased because women grouped in the intensive category have the highest rates of low birth weight (LBW). The objective of this paper is to provide a systematic examination of the Index and to uncover biases that may preclude its use in analyzing the association between resource utilization and birth outcomes. This is a cross-sectional study including all singleton live births in 1993 through 1996 (n = 591,403) in Ohio. Birth certificate data are used to derive the Index, which categorizes women as follows: Adequate Plus (A+), Adequate, Intermediate, and Inadequate. The Index is based on the ratio of observed to expected (O/E) number of prenatal visits. The expected number of visits is based on the American College of Obstetricians and Gynecologists (ACOG) recommendations. The Index also considers the month of initiation of prenatal care. The outcome measures are low birth weight (LBW) and small-for-gestational age (SGA). The LBW rate is 11.8% in the (A+) category, compared to 9.4% in the Inadequate category, and 3.3% and 3.5% in each of the Intermediate and Adequate categories, respectively. Preterm births are disproportionately represented in the (A+) category: 61.2% of births prior to 37 weeks are (A+), whereas only 18.9% of term births are (A+). This apparent bias results from the fact that the ACOG schedule of prenatal visits allocates nearly one third of the total visits to the last 4-5 weeks of gestation. A shorter gestational age implies fewer number of expected visits, a smaller denominator in the O/E ratio, and O/E ratios exceeding 100% by large margins. In fact, the observed number of visits exceeds the expected number of visits by only one or two in 40.1% of all births grouped in the (A+) category. Consequently, the Index yields misleading results indicating that women grouped in the (A+) category (or O/E ratios > 110%) are most likely to deliver LBW infants. Contrary to the results obtained through the APNCU Index, our gestational age-specific analysis showed that increasing number of prenatal visits is associated with improved birth outcomes. We recommend that the use of the APNCU Index to study the association between prenatal resource utilization and LBW be discontinued. PMID- 11864802 TI - Assessing health-related quality of life following myocardial infarction: is the SF-12 useful? AB - We report on the responsiveness of the SF-12 to changes in quality of life following acute myocardial infarction. Scores at 1, 6, 12, and 24 weeks postdischarge were compared with pre-MI health. Statistically significant differences and standardized response means were examined. Results were compared with the SF-36 subscales and previous reports. Respondents (n = 65) reported the expected poorer physical health at every follow-up, while expected changes in emotional health were observed at 6 but not 24 weeks. Comparison with the SF-36 subscales showed that although the SF-12 reflected the expected pattern of physical health, the summary score obscured an important association between perceptions of general health and participation in usual activities. This information is relevant for developing and evaluating rehabilitation interventions and self-managed recovery following MI. The SF-12 scores obscure important distinctions between quality of life domains, and are therefore not recommended for use following acute MI. PMID- 11864803 TI - Validation of diagnoses of peptic ulcers and bleeding from administrative databases: a multi-health maintenance organization study. AB - The automated health plan data and data from medical chart abstractions from eight large health maintenance organizations were used to evaluate the positive predictive values (PPVs) of the International Classification of Diseases, 9th revision (ICD-9) codes for cases of peptic ulcers and upper gastrointestinal bleeding. Overall, 207 of 884 cases of peptic ulcers and upper gastrointestinal bleeding (23%) were confirmed by surgery, endoscopy, X-ray, or autopsy. The PPVs were 66% for hospitalizations with codes for duodenal ulcer (ICD-9-CM 532), 61% for gastric/gastrojejunal ulcer (ICD-9-CM 531, 534), 1% for peptic ulcer (ICD-9 CM 533), and 9% for gastrointestinal hemorrhage (ICD-9-CM578). The overall and diagnostic category-specific PPVs were generally similar for the various HMOs. This study, using data from a large number of health plans located in different geographical regions, underscores the importance of evaluating the accuracy of the diagnoses from automated health plan databases. PMID- 11864804 TI - Benzodiazepine use and risk of dementia: a nested case-control study. AB - The objective of this article was to examine the possible association between benzodiazepine use and the risk of dementia in the elderly. This was a nested case--control study set in community settings in Bordeaux area, France. The participants were a representative sample of 3,777 elderly persons (65 years of age and older) followed from 1989 to 1997. The main outcome measures were the use of benzodiazepines in incident cases of dementia versus nondemented controls. On the basis of medical and psychological data, 150 patients were diagnosed with dementia according to the criteria of the third revision of the Diagnostic and Statistical Manual of Mental Disorders. Information on benzodiazepine use was obtained by face-to-face interview and visual assessment of patient's medicine chest by a trained neuropsychologist. After controlling for age, gender, education level, living alone, wine consumption, psychiatric history, and depressive symptomatology, ever use of benzodiazepines was associated with a significantly increased risk of dementia [adjusted odds ratio (OR), 1.7; 95% confidence interval, 1.2-2.4]. Former use was associated with a significantly increased risk of dementia (adjusted OR, 2.3; 95% CI,1.2-4.5). No association was found between dementia and the current use of benzodiazepines (adjusted OR, 1.0; 95% CI, 0.6-1.6). Our finding suggest that former use of benzodiazepines could be a risk factor for dementia, but more detailed investigation are needed. PMID- 11864805 TI - The ventral lateral posterior nucleus of the thalamus in schizophrenia: a post mortem study. AB - The ventral lateral posterior thalamic nucleus (VLp) is an integral part of both the cerebello-thalamocortical and the basal ganglia-thalamocortical circuit. Although both circuits are thought to be involved in the pathophysiology of schizophrenia, the VLp has not yet been examined in schizophrenia. Using stereological techniques in the brains of eight patients with schizophrenia and in eight age- and sex-matched controls, we measured the nuclear volume of the VLp and obtained estimates of the total number of neurons in this nucleus. Whole brain volume did not differ between the schizophrenia group and the control group and was not correlated to the volume of the right VLp or left VLp. The volume (minus sign25%) and the total neuron number (minus sign27%) of the left VLp were significantly reduced in the schizophrenia group. There were no significant differences in the nuclear volume, neuron density and total neuron number in the right VLp between the schizophrenia group and the control group. There were no significant correlations between length of illness and the nuclear volume, neuron density and total neuron number of the left and right VLp. The present results suggest that the total neuron number of the left VLp is reduced in the schizophrenia group, which may reflect disturbed cerebello-thalamocortical and basal ganglia-thalamocortical circuits in this disease. PMID- 11864806 TI - A transcranial magnetic stimulation study of inhibitory deficits in the motor cortex in patients with schizophrenia. AB - It has been proposed that inhibitory deficits play a crucial role in the pathophysiological process of schizophrenia as suggested by post-mortem, neuropsychological and neurophysiological evidence. We hypothesised that patients with schizophrenia would demonstrate abnormalities of cortical inhibition in the motor cortex with single and paired pulse transcranial magnetic stimulation (TMS). Patients with DSM-IV schizophrenia (n=22) and normal volunteers (n=21) participated in the study. Electromyographic recordings from the abductor pollicis brevis (APB) muscle were made during focal TMS stimulation to the contra lateral motor cortex. The threshold intensity to produce a motor response, the size of the motor evoked potential, the duration of the silent period, and the cortical inhibition and facilitation to paired pulse TMS were measured. The patient group demonstrated a reduction in length of the silent period and a reduction in cortical inhibition with paired stimuli. No changes were found in motor threshold, motor evoked potential size, or cortical facilitation. The study demonstrated deficits of cortical inhibition in the motor cortex of patients with schizophrenia. These deficits appear to be of cortical origin. Their relationship to dysfunction in other cortical networks requires further elucidation. PMID- 11864807 TI - Age, sex and laterality effects on cerebral glucose metabolism in healthy adults. AB - Normal cerebral glucose metabolism (CMRglc) was assessed with positron emission tomography in 66 healthy adults (28 women, 38 men; mean age 39, range 20--69 years) to determine effects of age, sex and laterality on CMRglc using statistical parametric mapping. Significant age-related decreases in global metabolism (gCMRglc) were noted in the entire sample and in both sexes, as well as widespread and bilateral decreases in cortical absolute regional metabolism (rCMRglc) and more focal anterior paralimbic normalized rCMRglc. However, significant positive correlations of age with normalized rCMRglc were observed in cerebellum, thalamus and occipital areas. Although the declines in gCMRglc and rCMRglc with age did not significantly differ between sexes, men compared with women had significantly lower gCMRglc and widespread decreased cortical and subcortical absolute rCMRglc. In the entire sample, and similarly in both sexes, left greater than right asymmetry was observed in medial frontal gyrus, posterior thalamus, lingual gyrus, cuneus and superior cingulate. The opposite laterality appeared in mesio-anterior cerebellum, and lateral frontal and temporal regions. Few regions showed significant interactions of metabolic laterality with either age or sex. These findings contribute toward a convergence in the literature, and the regression models of CMRglc vs. age serve as a normative database to which patients may be compared. PMID- 11864808 TI - Changes in brain functional homogeneity in subjects with Alzheimer's disease. AB - Imaging studies have reported marked reductions in brain glucose metabolism in Alzheimer's Disease (AD). However, less is known about disruptions in the patterns of brain metabolic activity. Here we questioned whether AD affects the patterns of homogeneity/heterogeneity in brain metabolism. PET images of 35 AD subjects were compared with those of 35 controls. A template was applied to extract a cortical rim, which was partitioned into 990 contiguous regions. Estimates of metabolic homogeneity were obtained using the coefficient of variation (CV). The CV of the entire cortex was found to be significantly larger in AD, suggesting increased heterogeneity at the whole brain level. In contrast, regional CV was significantly lower in AD in temporal and parietal cortices, which were the regions that along with the precuneus had the largest metabolic decrements, though the precuneus had increased CV. The enhanced heterogeneity for the global cortical pattern most likely reflects variability in the degree of pathology among brain regions as well as neuroanatomical disconnection. The enhanced homogeneity in parietal and temporal cortices is likely to reflect loss of regional differentiation (i.e. macrocolumnar disorganization). The enhanced CV in precuneus, despite its marked reductions in metabolism, suggests that increases in regional homogeneity in parietal and temporal cortices are not a mere reflection of the decrement in metabolism. PMID- 11864809 TI - Increased presence of white matter hyperintensities in adolescent patients with bipolar disorder. AB - Several reports have noted an increase in white matter hyperintensities (WMH) on MRI scans of adult patients with bipolar disorder. We investigated whether this increase was also evident in a group of adolescent patients with bipolar disorder. The sample consisted of 15 bipolar patients, 19 patients with schizophrenia and 16 healthy comparison subjects. All subjects were adolescents. WMH were blindly rated on T2-weighted and PD-weighted MRI scans using our own scale with documented inter-rater reliability. WMH were present in 10 of 15 bipolar patients (67%), seven of 19 patients with schizophrenia (37%) and five of 16 comparison subjects (31%). The bipolar adolescent group had a statistically significant increased presence of WMH compared both with healthy comparison subjects and the schizophrenic group. The association between WMH and bipolar disorder appears to extend to the adolescent years. PMID- 11864810 TI - Anti-apoptotic protein kinase of herpes simplex virus. PMID- 11864811 TI - Bacteria killing their own kind: novel bacteriocins of Pseudomonas and other gamma-proteobacteria. PMID- 11864812 TI - New approaches and old problems in the shadow of bioterrorism. PMID- 11864813 TI - Split personalities. PMID- 11864814 TI - Polarized secretion. PMID- 11864816 TI - Crowd control: quorum sensing in Enterococcus. PMID- 11864815 TI - Soil amoebae help to unravel fungal pathogenesis. PMID- 11864817 TI - Unraveling the host immune response to vacuolar pathogens. PMID- 11864819 TI - Quorum sensing in Pseudomonas aeruginosa: yet another player. PMID- 11864820 TI - What are high-temperature bacteria doing in cold environments? PMID- 11864821 TI - Giardia: highly evolved parasite or early branching eukaryote? AB - The phylogeny of the commonest protozoal agent of intestinal disease, Giardia, is unclear. Although recent intensive research suggests this important human parasite is an early branching eukaryote that evolved before the endosymbiotic origin of mitochondria, there is also evidence to suggest that, as a highly evolved parasite, it has lost many of its ancestral characteristics. In this case, these organisms might have arisen much more recently from aerobic free living flagellates. PMID- 11864822 TI - Dressed for success: the surface coats of insect-borne protozoan parasites. AB - Three major human diseases, malaria, sleeping sickness and leishmaniasis, are caused by protozoan parasites that are transmitted by blood-sucking insects. These insects are not mere 'flying syringes' that mechanically transfer parasites from one mammal to the next. Instead, they provide a specific environment--albeit not a particularly hospitable one--in which the parasites differentiate, proliferate and migrate to the correct tissues to ensure transmission to the next mammalian host. Recent studies on the role of parasite surface molecules in insect vectors have delivered some surprises and could provide insights on ways to interrupt transmission. PMID- 11864823 TI - Cyanobacterial photosynthesis in the oceans: the origins and significance of divergent light-harvesting strategies. AB - Prochlorococcus and Synechococcus are abundant unicellular cyanobacteria and major participants in global carbon cycles. Although they are closely related and often coexist in the same ocean habitat, they possess very different photosynthetic light-harvesting antennas. Whereas Synechococcus and the majority of cyanobacteria use phycobilisomes, Prochlorococcus has evolved to use a chlorophyll a(2)/b(2) light-harvesting complex. Here, we present a scenario to explain how the Prochlorococcus antenna might have evolved in an ancestral cyanobacterium in iron-limited oceans, resulting in the diversification of the Prochlorococcus and marine Synechococcus lineages from a common phycobilisome containing ancestor. Differences in the absorption properties and cellular costs between chlorophyll a(2)/b(2) and phycobilisome antennas in extant Prochlorococcus and Synechococcus appear to play a role in differentiating their ecological niches in the ocean environment. PMID- 11864824 TI - Hijacking the host: survival of pathogenic mycobacteria inside macrophages. AB - Mycobacteria are among the most persistent pathogens known today: one-third of the global population is latently infected with Mycobacterium tuberculosis, an organism that is responsible for approximately 2.5 million deaths each year. One key to the pathogenic potential of the mycobacteria lies in their capacity to resist destruction by macrophages. Although it has long been recognized that mycobacteria achieve such resistance by interfering with phagosome--lysosome fusion, recent work has shed some more light on the molecular basis of this highly efficient survival strategy. PMID- 11864825 TI - Cytolethal distending toxin: limited damage as a strategy to modulate cellular functions. AB - The coevolution of bacterial pathogens and their hosts has contributed to the development of very complex and sophisticated functional pathogen--host interfaces. Thus, well-adapted pathogens have evolved a variety of strategies to manipulate host cell functions precisely. For example, a group of unrelated Gram negative pathogenic bacteria have evolved a toxin, known as cytolethal distending toxin (CDT), that has the ability to control cell cycle progression in eukaryotic cells. Recent studies have identified CdtB as the active subunit of the CDT holotoxin. Through its nuclease activity, CdtB causes limited DNA damage, thereby triggering the DNA-damage response that ultimately results in the observed arrest of the cell cycle. In addition, it has been established that CDT is a tripartite AB toxin in which CdtB is the active 'A' subunit and CdtA and CdtC constitute the heterodimeric 'B' subunit required for the delivery of CdtB into the target cell. The mechanism of action of CDT suggests that the infliction of limited damage could be a strategy used by pathogenic bacteria to modulate host cell functions. PMID- 11864827 TI - The role of BAFF in B-cell maturation, T-cell activation and autoimmunity. PMID- 11864828 TI - Complement component C1 and the collectins--first-line defense molecules in innate and acquired immunity. PMID- 11864829 TI - Immunology and multimodal system interactions in health and disease. PMID- 11864830 TI - Dichotomy of glucocorticoid action in the immune system. PMID- 11864835 TI - IL-7 and homeostasis of recent thymic emigrants. PMID- 11864836 TI - Dangers of therapeutic manipulation of the Th-Th2 balance. PMID- 11864837 TI - Importance of the state of activation and/or differentiation of CD4+ T cells in AIDS pathogenesis. PMID- 11864840 TI - Heat-shock proteins as activators of the innate immune system. AB - Peptides bound or linked to heat-shock proteins (HSPs) of microbial or mammalian origin have been shown to elicit potent antigen-specific immunity. Some members of the HSP family, such as hsp60, hsp70, hsp90 and gp96, are able also to stimulate cells of the innate immune system directly and thus, act as 'danger' signaling molecules. This effect is independent of HSP-associated peptides and, in many respects, resembles the effect of lipopolysaccharide (LPS). Here, we discuss the similarities between the responses to HSPs and LPS and also, emphasize that care must be taken when working with preparations of HSPs in experimental settings and interpreting experimental data. PMID- 11864841 TI - Does the shape of lipid A determine the interaction of LPS with Toll-like receptors? AB - Lipopolysaccharide (LPS) triggers the activation of the immune system through the induction of cytokine release. Although it was assumed initially that LPS molecules from different bacteria are similar, recent evidence suggests that structural and functional differences between LPS species are the rule rather than the exception. It has been proposed that the shape of the lipid A component determines the bioactivity of LPS, with lipid A that adopts a conical conformation being more active than lipid A that adopts a cylindrical shape. The mechanism linking the molecular conformation with the biological activity of LPS has not been elucidated. We propose that LPS with a conical shape (e.g. from Escherichia coli) induces cytokine production through Toll-like receptor 4 (TLR4), whereas more cylindrical LPS (e.g. from Porphyromonas gingivalis) induces expression of a different set of cytokines through TLR2. Strictly cylindrical LPS molecules (e.g. the lipid A precursor Ia or from Rhodobacter sphaeroides) have antagonistic properties at the level of TLRs. PMID- 11864842 TI - Cbl and Cbl-b in T-cell regulation. AB - Genetic studies indicate that Cbl and Cbl-b, two highly homologous adaptor proteins, are involved in the negative regulation of thymocyte development and peripheral T-cell activation, respectively. The recent identification of Cbl proteins as RING-type E3 ubiquitin ligases might provide insights into their distinct immune regulatory functions, involving the targeting of different substrates for ubiquitination. The structural similarity and ubiquitous expression of Cbl and Cbl-b suggest, however, that they might also have overlapping functions in setting the thresholds for thymocyte selection and mature T-cell signaling. PMID- 11864843 TI - Prostaglandins as modulators of immunity. AB - Prostaglandins are potent lipid molecules that affect key aspects of immunity. The original view of prostaglandins was that they were simply immunoinhibitory. This review focuses on recent findings concerning prostaglandin E2 (PGE2) and the PGD2 metabolite 15-deoxy-Delta(12,14)-PGJ2, and their divergent roles in immune regulation. We will highlight how these two seminal prostaglandins regulate immunity and inflammation, and play an emerging role in cancer progression. Understanding the diverse activities of these prostaglandins is crucial for the development of new therapies aimed at immune modulation. PMID- 11864844 TI - The role of mast cells in host defense and their subversion by bacterial pathogens. AB - Mast cells (MCs) play a prominent role in the early immune response to invading pathogenic bacteria. This newly discovered role for MCs involves the release of chemoattractants that recruit neutrophils and the direct phagocytosis and killing of opsonized bacteria. Whereas these activities are clearly beneficial to the host, certain pathogens have evolved mechanisms to evoke anomalous MC responses to the detriment of the host. These include evoking phagocytosis without killing of unopsonized bacteria and the production of toxins that corrupt the release of mediators by MCs. Elucidating how pathogens subvert the activities of MCs could provide clues to limiting the pathological activities of these cells during infectious diseases. PMID- 11864845 TI - Caveolae and caveolin in immune cells: distribution and functions. AB - Caveolae are small, cholesterol-rich, hydrophobic membrane domains, characterized by the presence of the protein caveolin and involved in several cellular processes, including clathrin-independent endocytosis, the regulation and transport of cellular cholesterol, and signal transduction. Recently, caveolae have been identified as providing a novel route by which several pathogens are internalized by antigen-presenting cells and as centers for signal transduction. Here, we review the distribution and role of caveolae and caveolin in mammalian immune cells. PMID- 11864846 TI - Towards a European nonhuman primate immune-tolerance network. PMID- 11864848 TI - Let's get serious about promoting physical activity. PMID- 11864849 TI - Parental feeding behavior and children's fat mass. PMID- 11864850 TI - Body-composition assessment via air-displacement plethysmography in adults and children: a review. AB - Laboratory-based body-composition techniques include hydrostatic weighing (HW), dual-energy X-ray absorptiometry (DXA), measurement of total body water (TBW) by isotope dilution, measurement of total body potassium, and multicompartment models. Although these reference methods are used routinely, each has inherent practical limitations. Whole-body air-displacement plethysmography is a new practical alternative to these more traditional body-composition methods. We reviewed the principal findings from studies published between December 1995 and August 2001 that compared the BOD POD method (Life Measurement, Inc, Concord, CA) with reference methods and summarized factors contributing to the different study findings. The average of the study means indicates that the BOD POD and HW agree within 1% body fat (BF) for adults and children, whereas the BOD POD and DXA agree within 1% BF for adults and 2% BF for children. Few studies have compared the BOD POD with multicompartment models; those that have suggest a similar average underestimation of approximate 2-3% BF by both the BOD POD and HW. Individual variations between 2-compartment models compared with DXA and 4 compartment models are partly attributable to deviations from the assumed chemical composition of the body. Wide variations among study means, -4.0% to 1.9% BF for BOD POD - HW and -3.0% to 1.7% BF for BOD POD - DXA, are likely due in part to differences in laboratory equipment, study design, and subject characteristics and in some cases to failure to follow the manufacturer's recommended protocol. Wide intersubject variations between methods are partly attributed to technical precision and biological error but to a large extent remain unexplained. On the basis of this review, future research goals are suggested. PMID- 11864851 TI - Resting metabolic rate, plasma leptin concentrations, leptin receptor expression, and adipose tissue measured by whole-body magnetic resonance imaging in women with Prader-Willi syndrome. AB - BACKGROUND: Obesity in Prader-Willi syndrome (PWS) may be related to abnormalities in the adipocyte-leptin-hypothalamic pathway and may be exacerbated by reductions in the resting metabolic rate (RMR). OBJECTIVE: We compared body composition, body-composition- adjusted RMR, and adiposity-adjusted plasma leptin between women with PWS and control women. We also examined leptin receptor expression in the PWS group. DESIGN: We studied body composition using whole-body magnetic resonance imaging and measured plasma leptin by radioimmunoassay in 45 control women aged 18-56 y and in 13 women with PWS aged 20-38 y. RMR was measured by indirect calorimetry in 41 control women and in 8 women with PWS. Age, body composition, and regional adipose tissue (AT) depots were corrected for by multiple regression analysis. Messenger RNA expression of the leptin receptor was examined by reverse transcriptase-polymerase chain reaction in lymphocytes. RESULTS: In the PWS group, fat mass was greater after correction for fat-free mass, and RMR was normal after correction for both fat-free mass and fat mass. Leptin was influenced primarily by subcutaneous AT volume in both subject groups. Leptin concentrations were not significantly different between the 2 groups after adjustment for age and AT content or distribution. Full-length leptin receptor messenger RNA was expressed in the lymphocytes of the PWS group. CONCLUSIONS: Differences in RMR in women with PWS are explained by abnormal body composition, suggesting that energy expenditure is normal at the tissue level in PWS. There is no evidence that defective leptin production causes obesity in PWS, and leptin receptor deficiency is not a primary consequence of the gene defects leading to leptin resistance. PMID- 11864852 TI - Eating behavior correlates of adult weight gain and obesity in healthy women aged 55-65 y. AB - BACKGROUND: The specific underlying causes of adult weight gain remain uncertain. OBJECTIVE: The objective was to determine the association of 3 measures of eating behavior with weight gain and body mass index (BMI; in kg/m(2)) in adults. DESIGN: Current dietary restraint, disinhibition, and hunger were assessed with the use of the Eating Inventory in 638 healthy, nonsmoking women aged 55-65 y. In addition, subjects reported their current weight and height, their weight for 6 age intervals, and changes in voluntary dietary energy restriction over the past 10 y. Current weight and height were validated in 10% of subjects. RESULTS: Current disinhibition strongly predicted weight gain and current BMI (partial r = 0.27 and 0.34, respectively, both P < 0.001). Neither restraint nor hunger was a significant independent predictor of either variable, but the positive associations between disinhibition and both weight gain and BMI were attenuated by restraint (P = 0.016 and 0.010, respectively, after adjustment for confounding variables). In the subpopulation of women who reported a stable level of voluntary dietary energy restriction, disinhibition also strongly predicted weight gain and higher BMI, and restraint was negatively associated with weight gain (partial r = -0.17, P = 0.019). CONCLUSIONS: Higher disinhibition is strongly associated with greater adult weight gain and higher current BMI, and dietary restraint may attenuate this association when disinhibition is high. These findings suggest that eating behavior has an important role in the prevention of adult-onset obesity and that further studies are warranted. PMID- 11864853 TI - Fatty acid saturation of the diet and plasma lipid concentrations, lipoprotein particle concentrations, and cholesterol efflux capacity. AB - BACKGROUND: The fatty acid content and saturation degree of the diet can modulate HDL composition and cholesterol efflux. OBJECTIVE: We studied the modifications in plasma lipoprotein particles and serum capacity to stimulate cholesterol efflux induced by different fatty acids. DESIGN: Seventeen women and 24 men followed in the same sequence 4 diets containing 35% of total energy as fat. The saturated fat diet contained 17% palm oil; the monounsaturated fat diet, 20.9% olive oil; the n-6 polyunsaturated fat diet, 12.5% sunflower oil; and the n-3 polyunsaturated fat diet, sunflower oil supplemented with 4-4.5 g fish oil/d. Each phase lasted 4-5 wk. RESULTS: In both sexes, apolipoprotein (apo) A-I concentrations were significantly lower with unsaturated fat diets than with the saturated fat diet, but concentrations of lipoproteins containing only apo A-I (Lp A-I) were lower only in the men. Concentrations of lipoproteins containing both apo A-I and apo A-II (Lp A-I:A-II) were lower with both polyunsaturated fat diets in the women but significantly higher in the men. Lp E concentrations were significantly higher with the 2 polyunsaturated fat diets. Lp E non-B particle concentrations were not modified in the men but were significantly higher in the women in both polyunsaturated fat phases. Lp C-III concentrations were higher with the saturated fat diet only in the men. The serum samples taken after the n 3 polyunsaturated fat phase were the most efficient for extracting cellular cholesterol in both sexes. CONCLUSIONS: The monounsaturated and polyunsaturated fat diets were healthier, producing a better lipid profile. The n-3 polyunsaturated fat diet increased the capacity of serum to promote the efflux of cholesterol from cells in culture. PMID- 11864854 TI - Relation between a diet with a high glycemic load and plasma concentrations of high-sensitivity C-reactive protein in middle-aged women. AB - BACKGROUND: Recent prospective data suggest that intake of rapidly digested and absorbed carbohydrates with a high dietary glycemic load is associated with an increased risk of ischemic heart disease. OBJECTIVE: We examined whether a high dietary glycemic load was associated with elevated hs-CRP concentrations and whether this association was modified by body mass index (BMI; in kg/m(2)). DESIGN: In 244 apparently healthy women, we measured plasma hs-CRP concentrations and determined average dietary glycemic loads with a validated semiquantitative food-frequency questionnaire. Using multiple regression models, we evaluated the association between dietary glycemic load and plasma hs-CRP after adjusting for age; treatment status; smoking status; BMI; physical activity level; parental history of myocardial infarction; history of hypertension, diabetes, and high cholesterol; postmenopausal hormone use; alcohol intake; and other dietary variables. RESULTS: We found a strong and statistically significant positive association between dietary glycemic load and plasma hs-CRP. The median hs-CRP concentration for the lowest quintile of dietary glycemic load was 1.9 mg/L and for the highest quintile was 3.7 mg/L; corresponding multivariate-adjusted geometric means were 1.4 and 3.8 mg/L, respectively (P for trend < 0.01). This association was significantly modified by BMI. Among women with a BMI greater than-or-equal 25, the multivariate-adjusted geometric mean hs-CRP concentration in the lowest quintile was 1.6 mg/L and in the highest quintile was 5.0 mg/L; however, among women with a BMI < 25, the corresponding means were 1.1 and 3.1 mg/L, respectively (P = 0.01 for interaction). CONCLUSIONS: Dietary glycemic load is significantly and positively associated with plasma hs-CRP in healthy middle aged women, independent of conventional risk factors for ischemic heart disease. Exacerbation of the proinflammatory process may be a mechanism whereby a high intake of rapidly digested and absorbed carbohydrates increases the risk of ischemic heart disease, especially in overweight women prone to insulin resistance. PMID- 11864855 TI - Free-living activity energy expenditure in women successful and unsuccessful at maintaining a normal body weight. AB - BACKGROUND: Although physical inactivity is believed to contribute to the rising prevalence of obesity, the role and magnitude of its contribution to weight gain are unknown. OBJECTIVE: We compared total free-living activity energy expenditure (AEE) and physical activity level in women successful and unsuccessful at maintaining a normal body weight. DESIGN: Premenopausal, generally sedentary women were studied at their normal weight and 1 y later after no intervention. Two groups were identified on the basis of extreme weight changes: maintainers (n = 27) had a weight gain of less-than-or-equal 3% of their initial body weight ( less-than-or-equal 2 kg/y) and gainers (n = 20) had a weight gain of >10% (>6 kg/y). At baseline and follow-up, evaluations were conducted during 4 wk of diet controlled, energy-balance conditions. Free-living AEE and physical activity were assessed with the use of doubly labeled water, exercise energy economy and muscle strength with the use of standardized exercise tests, and sleeping EE and substrate utilization with the use of chamber calorimetry. RESULTS: Maintainers lost a mean (plus minusSD) of 0.5 plus minus 2.2 kg/y and gainers gained 9.5 plus minus 2.1 kg/y. Gainers had a lower AEE (P < 0.02), a lower physical activity level (P < 0.01), and less muscle strength (P < 0.001); these differences between groups remained significant from baseline to follow-up. Sleeping EE, exercise economy, and sleeping or 24-h substrate utilization were not significantly different between the 2 groups. A lower AEE in the gainers explained approximately 77% of their greater weight gain after 1 y. CONCLUSION: The general US population should increase their daily physical activity levels to decrease the rising prevalence of obesity. PMID- 11864857 TI - Endogenous glycine and tyrosine production is maintained in adults consuming a marginal-protein diet. AB - BACKGROUND: The adequacy of indispensable amino acid supplies has received much attention in studies of protein requirements, but the availability of nitrogen for synthesis and maintenance of the supply of dispensable amino acids has been overlooked. OBJECTIVE: We aimed to determine whether nitrogen balance and the endogenous supply of the dispensable amino acids glycine and tyrosine can be maintained with a marginal protein intake. DESIGN: Phenylalanine, glycine, and tyrosine kinetics were measured in young adults (6 men, 6 women) on 4 occasions during a reduction in habitual protein intake (1.13 g x kg(-1) x d(-1)) to a marginal intake (0.75 g x kg(-1) x d(-1)) by using a multiple stable-isotope infusion protocol. RESULTS: During the 10-d period of marginal protein intake, nitrogen excretion fell initially, then remained constant such that nitrogen balance was negative for the first 2 d and then positive or zero thereafter. Whole-body protein degradation and synthesis predicted from phenylalanine kinetics declined significantly (P < 0.05) over the period of marginal protein intake. Despite the reduction in the amount of glycine and tyrosine derived from whole-body proteolysis, the fluxes of glycine and tyrosine were maintained. CONCLUSIONS: The results show that adaptation to a marginal intake of dietary protein consisted of an overall reduction in whole-body protein turnover, net protein catabolism, and the rate of nitrogen excretion. The conserved nitrogen was sufficient to maintain the endogenous synthesis and hence the supply of glycine and tyrosine. PMID- 11864856 TI - Extended effects of evening meal carbohydrate-to-fat ratio on fasting and postprandial substrate metabolism. AB - BACKGROUND: High-fat and high-carbohydrate diets lead to insulin resistance, gastrointestinal adaptation, and high plasma triacylglycerol concentrations. It is unclear, however, how rapidly these changes occur. OBJECTIVE: We sought to determine the effects of both high-fat and high-carbohydrate evening meals on parameters of insulin resistance, hypertriglyceridemia, and gastrointestinal hormones. DESIGN: Twelve healthy men were studied on 4 separate occasions. On 2 occasions, the subjects received a high-fat evening meal (62% of energy from fat) and on the other 2 occasions the subjects received a low-fat evening meal (16% of energy from fat). The morning after each meal the subjects were administered either an oral-fat-tolerance test or an oral-glucose-tolerance test. Plasma samples were analyzed for glucose, insulin, fatty acids, 3-hydroxybutyrate, triacylglycerol, pancreatic polypeptide, peptide YY, and cholecystokinin. Postchallenge data were analyzed by two-way analysis of variance with interaction and fasting concentrations analyzed by repeated-measures analysis of variance. RESULTS: Fasting plasma concentrations of triacylglycerol were significantly elevated 12 h after each evening meal, but fatty acid and 3-hydroxybutyrate concentrations were reduced. No effects on glucose or insulin concentrations were detected. The high-fat evening meals elevated plasma cholecystokinin concentrations, reduced fasting concentrations of pancreatic polypeptide, and had no significant effect on peptide YY concentrations. The ratio of fat to carbohydrate in the evening meal produced significant effects on plasma triacylglycerol and fatty acids during both the oral-fat-tolerance and oral glucose-tolerance tests. CONCLUSIONS: The present study showed that the effects of high-fat and high-carbohydrate evening meals persist at least overnight and suggests that knowledge of recent dietary history is essential to the effective design of metabolic studies. PMID- 11864858 TI - Comparison of energy expenditure estimates from doubly labeled water, a physical activity questionnaire, and physical activity records. AB - BACKGROUND: Various methods are used by epidemiologists to estimate the energy cost of physical activity; these include physical activity records and recalls. However, there is limited validation of these methods against the doubly labeled water technique for determining energy expenditure (EE). OBJECTIVE: We compared EE as estimated by indirect methods (physical activity records and recall questionnaires) used in epidemiologic studies with EE obtained from doubly labeled water (EE(DLW)) in free-living men. DESIGN: We determined EE(DLW), energy intake at weight maintenance, and EE from 7-d physical activity records (EE(Record)) and a 7-d physical activity recall questionnaire (EE(Recall)) in 24 men aged 41 plus minus 2.0 y ( plus minus SEM) with a body mass index (in kg/m(2)) of 25.1 plus minus 0.5. RESULTS: There was excellent agreement between EE(DLW) (13.27 plus minus 0.35 MJ/d) and energy intake (13.19 plus minus 0.36 MJ/d), with a difference of 0.5 plus minus 1.0% ( plus minus SE). The indirect measures of physical activity and EE were 14.17 plus minus 0.37 MJ/d for EE(Record) (difference from EE(DLW): 7.9 plus minus 3.2%) and 17.40 plus minus 1.45 MJ/d for EE(Recall) (difference from EE(DLW): 30.6 plus minus 9.9%). CONCLUSIONS: Seven-day physical activity records provide an acceptable estimate of EE in free-living adults compared with EE(DLW), but 7-d physical activity recalls have limited application to estimate daily EE. For optimal validity, the 7-d physical activity records require good subject compliance and the provision of careful instructions for their use. PMID- 11864859 TI - Vegetable-borne lutein, lycopene, and beta-carotene compete for incorporation into chylomicrons, with no adverse effect on the medium-term (3-wk) plasma status of carotenoids in humans. AB - BACKGROUND: The results of epidemiologic studies have consistently shown associations between dietary intake or plasma carotenoid status and incidence of cancers and cardiovascular and eye diseases. OBJECTIVE: The aim was to assess whether vegetable-borne carotenoids (lycopene, lutein, and beta-carotene) compete for intestinal absorption and whether this affects the plasma status of carotenoids in the medium term (ie, after 3 wk). DESIGN: During 3-wk periods separated by 3-wk washout periods, 20 women were supplemented with either 96 g tomato puree/d (14.98 mg lycopene + 1.50 mg beta-carotene), 92 g cooked chopped spinach/d (11.93 mg lutein + 7.96 mg beta-carotene), 96 g tomato puree/d + 92 g chopped spinach/d, 96 g tomato puree/d + 2 lutein pills (12 mg lutein), or 92 g chopped spinach/d + 1 lycopene pill (15 mg lycopene). Plasma carotenoids were measured before and after each supplementation period. The subjects also participated in postprandial experiments in which they ingested meals containing double amounts of the supplements described above. Carotenoids were measured in chylomicrons to assess the interaction of carotenoids on absorption. RESULTS: Adding a second carotenoid to a meal that provided a first carotenoid diminished the chylomicron response to the first carotenoid. However, cosupplementation with a second carotenoid of a diet supplemented with a first carotenoid did not diminish the medium-term plasma response to the first carotenoid. CONCLUSION: Consumption of carotenoids from different vegetable sources does not diminish plasma carotenoid concentrations in the medium term, despite the finding in postprandial testing of competitive inhibitory interactions among different carotenoids. PMID- 11864860 TI - Iron bioavailability from iron-fortified Guatemalan meals based on corn tortillas and black bean paste. AB - BACKGROUND: Corn masa flour is widely consumed in Central America and is therefore a potentially useful vehicle for iron fortification. OBJECTIVE: The goal was to evaluate the bioavailability of iron from meals based on corn tortillas and black bean paste that were fortified with ferrous fumarate, ferrous sulfate, or NaFeEDTA and to investigate the potential of Na(2)EDTA to increase the bioavailability of iron from ferrous fumarate. DESIGN: With use of a crossover study design, iron bioavailability was measured in Guatemalan girls aged 12-13 y by a stable-isotope technique based on erythrocyte incorporation 14 d after intake. RESULTS: Geometric mean iron bioavailability from test meals fortified with ferrous fumarate was 5.5-6.2% and was not improved significantly by the addition of Na(2)EDTA at molar ratios of 1:1 relative to fortification iron or to the total iron content of the fortified corn masa flour. Geometric mean iron bioavailability from test meals fortified with ferrous sulfate was 5.5% and was significantly higher in test meals fortified with NaFeEDTA (9.0%; P = 0.009, paired t test). CONCLUSIONS: The bioavailability of iron from ferrous fumarate was not improved by the addition of Na(2)EDTA, contrary to what was previously shown for ferrous sulfate in other cereal-based meals. However, the bioavailability of iron from the test meal was significantly enhanced when NaFeEDTA replaced ferrous sulfate. These results support the use of NaFeEDTA in the fortification of inhibitory staple foods such as corn masa flour. PMID- 11864862 TI - Low dietary magnesium increases supraventricular ectopy. AB - BACKGROUND: Magnesium has been suggested to be beneficial in counteracting all phases of the processes that lead to ischemic heart disease, including terminal events such as arrhythmia and sudden death. OBJECTIVE: We tested the hypothesis that an intake of magnesium considerably below the recommended dietary allowance can produce chemical and physiologic evidence of depletion. DESIGN: Twenty-two postmenopausal women were maintained in a metabolic unit and ate a diet of conventional foods containing less than one-half of or more than the recommended dietary allowance for magnesium (320 mg/d). Dietary assignments were random and double blind in a crossover design. Magnesium concentrations were measured by spectroscopy and ion-specific electrolyte analysis, and Holter electrocardiograms lasting approximate 21 h were recorded. RESULTS: Magnesium concentrations in erythrocytes, serum (total and ultrafilterable), and urine were significantly lower when dietary magnesium was lower. Holter monitors showed a significant increase in both supraventricular and supraventricular plus ventricular beats when the dietary magnesium concentration was low. Hypomagnesemia, hypocalcemia, and hypokalemia were not found. CONCLUSIONS: The magnesium requirement was defined with the use of biochemical and electrophysiologic criteria. The recommended dietary allowance of 320 mg/d seems correct; 130 mg is too little. Persons who live in soft water areas, who use diuretics, or who are predisposed to magnesium loss or ectopic beats may require more dietary magnesium than would others. PMID- 11864861 TI - Long-term intake of vitamins and carotenoids and odds of early age-related cortical and posterior subcapsular lens opacities. AB - BACKGROUND: Proper nutrition appears to protect against cataracts. Few studies have related nutrition to the odds of developing cortical or posterior subcapsular (PSC) cataracts. OBJECTIVE: We assessed the relation between usual nutrient intakes and age-related cortical and PSC lens opacities. DESIGN: We studied 492 nondiabetic women aged 53-73 y from the Nurses' Health Study cohort who were without previously diagnosed cataracts. Usual nutrient intake was calculated as the average intake from 5 food-frequency questionnaires collected over a 13-15-y period before the eye examination. Duration of vitamin supplement use was determined from 7 questionnaires collected during this same period. We defined cortical opacities as grade > or = 0.5 and subcapsular opacities as grade > or =0.3 of the Lens Opacities Classification System III. RESULTS: Some lenses had more than one opacity. No nutrient measure was related to prevalence of opacities in the full sample, but significant interactions were seen between age and vitamin C intake (P = 0.02) for odds of cortical opacities and between smoking status and folate (P = 0.02), alpha-carotene (P = 0.02), beta-carotene (P = 0.005), and total carotenoids (P = 0.02) for odds of PSC opacities. For women aged <60 y, a vitamin C intake > or = 362 mg/d was associated with a 57% lower odds ratio (0.43; 95% CI: 0.2, 0.93) of developing a cortical cataract than was an intake <140 mg/d, and use of vitamin C supplements for > or = 10 y was associated with a 60% lower odds ratio (0.40; 0.18, 0.87) than was no vitamin C supplement use. Prevalence of PSC opacities was related to total carotenoid intake in women who never smoked (P = 0.02). CONCLUSIONS: Our results support a role for vitamin C in diminishing the risk of cortical cataracts in women aged <60 y and for carotenoids in diminishing the risk of PSC cataracts in women who have never smoked. PMID- 11864863 TI - Incorporation of deuterated RRR- or all-rac-alpha-tocopherol in plasma and tissues of alpha-tocopherol transfer protein--null mice. AB - BACKGROUND: Most vitamin E supplements contain synthetic all-rac-alpha-tocopherol [2,5,7,8-tetramethyl-2RS-(4'RS,8'RS,12-trimethyltridecyl)-6-chromanol] with 8 stereoisomers; only 1 is identical to the natural stereoisomer, RRR-alpha tocopherol [2,5,7,8-tetramethyl-2R-(4'R,8'R,12-trimethyltridecyl)-6-chromanol]. In humans, 2R-alpha-tocopherol stereoisomers are preferentially maintained in the plasma, a function that has been attributed to hepatic alpha-tocopherol transfer protein (alpha-TTP), but this hypothesis has not been tested. OBJECTIVE: We sought to determine the functions of alpha-TTP by comparing mice that express alpha-TTP with mice that are genetically unable to express alpha-TTP. DESIGN: Adult alpha-TTP null (Ttpa(-/-); n = 5), heterozygous (Ttpa(+/-); n = 7), and wild-type (Ttpa(+/+); n = 3) mice consumed equimolar RRR-alpha-[5,7 (C(2)H(3))(2)]-(d(6))- and all-rac-alpha-[5-(C(2)H(3))]-(d(3))-tocopheryl acetates (30 mg/kg diet each) for 3 mo. Subsequently, we measured labeled and unlabeled alpha-tocopherols in plasma and 17 tissues. RESULTS: In all mice, plasma and tissue d(6)- + d(3)-alpha-tocopherols represented approximate 80-90% of total alpha-tocopherol. In the Ttpa(-/-) mice, low total alpha-tocopherol concentrations were found in plasma (5.4%) and most other tissues (2-20%), but liver concentrations were 39% of those of Ttpa(+/+) mice. Peripheral tissue ratios of d(6)- to d(3)-alpha-tocopherol were 1.1 plus minus 0.1 and 1.8 plus minus 0.2 in Ttpa(-/-) and Ttpa(+/+) mice, respectively (P < 0.0001), showing that alpha-TTP preferentially selects 2R-alpha-tocopherols for secretion into plasma. This 2:1 ratio does not support the currently defined international unit of 1.36:1 RRR-alpha-tocopherol to all-rac-alpha-tocopherol. CONCLUSION: Deletion of the alpha-TTP gene in mice results in an accumulation of dietary alpha tocopherol in the liver and depletion of peripheral tissue alpha-tocopherol. PMID- 11864864 TI - Anthropometry-based reference values for 24-h urinary creatinine excretion during growth and their use in endocrine and nutritional research. AB - BACKGROUND: Urinary creatinine reference values that take anthropometric data into account, which is mandatory during growth, are not available for healthy white children. OBJECTIVE: We sought to establish anthropometry-based reference values for 24-h urinary creatinine excretion in healthy white children aged 3-18 y. DESIGN: Anthropometric variables and 24-h urinary creatinine excretion rates were determined cross-sectionally (225 boys and 229 girls). Age and sex dependency of 24-h creatinine excretion (crude and related to individual anthropometric variables) were assessed to derive appropriate creatinine reference values. The applicability of these creatinine reference values for estimation of daily excretion of certain analytes was assessed in 40 additional children. RESULTS: Sex-specific, body-weight-related creatinine reference values were derived for the following age groups: 3, 4-5, 6-8, 9-13, and 14-18 y. The 5th percentile exceeded 0.1 mmol x kg(-1) x d(-1) in all age groups >3 y. The use of these creatinine reference values for estimating average 24-h excretion rates of certain analytes (determined as the ratio of analyte to creatinine in spot urine samples) yielded reasonable estimates of mean 24-h urinary excretion rates actually analyzed (spot and 24-h urine samples from the same children). Ideal 24 h creatinine excretion values for height were also derived for a potential determination of the creatinine height index. CONCLUSIONS: Established anthropometry-based creatinine reference values are recommended as a convenient, simple tool to 1) identify severe 24-h urine collection errors, 2) calculate average 24-h excretion rates of certain analytes (from respective ratios of analyte to creatinine) determined in spot urine samples, and 3) assess somatic protein status by determining the creatinine height index. PMID- 11864865 TI - A randomized controlled trial of long-chain polyunsaturated fatty acid supplementation of formula in term infants after weaning at 6 wk of age. AB - BACKGROUND: The critical period during which the dietary supply of long-chain polyunsaturated fatty acids (LCPs) may influence the maturation of cortical function in term infants is unknown. OBJECTIVE: The aim of the present study was to determine the relative importance for maturation of the visual cortex of the dietary supply of LCPs during the first 6 wk of life compared with that during weeks 7-52. DESIGN: A randomized controlled clinical trial of LCP supplementation in 65 healthy term infants who were weaned from breast-feeding at 6 wk of age was conducted to determine whether the dietary supply of LCPs after weaning influenced the maturation of visual acuity and stereoacuity. RESULTS: Despite a dietary supply of LCPs from breast milk during the first 6 wk of life, infants who were weaned to formula that did not provide LCPs had significantly poorer visual acuity at 17, 26, and 52 wk of age and significantly poorer stereoacuity at 17 wk of age than did infants who were weaned to LCP-supplemented formula. Better acuity and stereoacuity at 17 wk was correlated with higher concentrations of docosahexaenoic acid in plasma. Better acuity at 52 wk was correlated with higher concentrations of docosahexaenoic acid in plasma and red blood cells. No significant effects of diet on growth were found. CONCLUSION: The results suggest that the critical period during which the dietary supply of LCPs can influence the maturation of cortical function extends beyond 6 wk of age. PMID- 11864866 TI - Relation between mothers' child-feeding practices and children's adiposity. AB - BACKGROUND: The prevalence of obesity in American children is currently estimated to be 25%. Poor nutritional habits during childhood have been directly related to pediatric obesity. OBJECTIVE: Our objective was to evaluate the relation between mothers' child-feeding practices and children's adiposity in a sample of boys and girls from 2 ethnic groups. DESIGN: A total of 74 white (25 boys and 49 girls) and 46 African American (22 boys and 24 girls) children ( plus minus SD age: 11 plus minus 1.7 y) and their mothers participated in this study. The children's body composition was assessed by dual-energy X-ray absorptiometry. The mothers' child-feeding practices were assessed with the Child Feeding Questionnaire. Dietary intake data were based on three 24-h dietary recalls conducted by use of the multiple-pass technique. RESULTS: Two subscales of the Child Feeding Questionnaire, pressure to eat and concern for child's weight, explained 15% of the variance in total fat mass in both African American and white boys and girls (P < 0.001) after correction for total lean mass and energy intake (which explained 5% of the variance in total fat mass). Ethnicity, sex, and socioeconomic status did not contribute significantly to variance in total fat mass. CONCLUSIONS: Child-feeding practices are key behavioral variables that explain more of the variance in total fat mass than does energy intake in a biethnic population of boys and girls. These findings have important implications for the prevention of obesity in children because they suggest that prevention programs need to focus on the feeding behaviors of parents in addition to the macronutrient and energy intakes of children. PMID- 11864867 TI - Use of a Durnin-Womersley formula to estimate change in subcutaneous fat content in HIV-infected subjects. AB - BACKGROUND: HIV-infected individuals may develop malnutrition or lipodystrophy, leading to losses of subcutaneous adipose tissue (SAT). OBJECTIVE: We compared the ability of a Durnin-Womersley formula for total adipose tissue (TAT) to estimate change in SAT with the use of whole-body magnetic resonance imaging (MRI) as a criterion measure. DESIGN: We analyzed data from 2 clinical trials: a prospective randomized trial of protein supplements, progressive resistance training, or combined treatment in 29 malnourished, HIV-positive women, and a prospective open-label trial of recombinant human growth hormone in 25 HIV infected subjects with visceral adipose tissue (VAT) accumulation. Changes in fat by the Durnin-Womersley formula and in SAT, TAT, and VAT by MRI were compared by linear regression, and Bland-Altman analyses were used to assess the agreement between the prediction and criterion methods. The repeatability of the Durnin Womersley measurement was evaluated in 14 weight-stable, healthy adults studied twice within 1 y. RESULTS: At baseline, Durnin-Womersley fat was significantly associated with SAT (r(2) = 0.75, P < 0.001) and TAT (r(2) = 0.79, P < 0.001) but not with VAT. Change in Durnin-Womersley fat was significantly associated with change in SAT (r(2) = 0.66, P < 0.001) and in TAT (r(2) = 0.57, P < 0.001) but not in VAT. The limits of agreement for the Durnin-Womersley estimation of change in SAT were -3.4 to 2.6 kg and the SEE was 1.5 kg. The SEE for repeated measures of SAT in healthy control subjects was 0.84. CONCLUSIONS: The Durnin-Womersley formula can be used to predict change in SAT. The limits of agreement and the SEE for predicting change in SAT by MRI are approximately twice as great as the error of repeated Durnin-Womersley measures in control subjects. PMID- 11864868 TI - Moderate alcohol consumption lowers risk factors for cardiovascular disease in postmenopausal women fed a controlled diet. AB - BACKGROUND: Moderate alcohol consumption (1-2 drinks/d) may decrease cardiovascular disease risk in postmenopausal women by improving lipid profiles. OBJECTIVE: We measured the effect of moderate alcohol consumption on lipids and lipoproteins in postmenopausal women. DESIGN: Postmenopausal women (n = 51) consumed 0 (control), 15 (1 drink), and 30 (2 drinks) g alcohol (ethanol)/d for 8 wk each as part of a controlled diet in a randomized crossover design. The control diet provided approximately 15%, 53%, and 32% of energy from protein, carbohydrate, and fat, respectively. The energy provided from alcohol in the 15- and 30-g alcohol diets was replaced with energy from carbohydrate. RESULTS: Compared with concentrations after the control diet, plasma LDL cholesterol decreased from 3.45 to 3.34 mmol/L (P = 0.04) and triacylglycerol from 1.43 to 1.34 mmol/L (P = 0.05) after 15 g alcohol/d. There were no additional significant decreases in either lipid after an increase in alcohol intake from 15 to 30 g/d. Compared with concentrations after the control diet, plasma HDL cholesterol increased nonsignificantly from 1.40 to 1.43 mmol/L after 15 g alcohol/d but increased to 1.48 mmol/L after 30 g alcohol/d (P = 0.02). Apolipoprotein A-I increased significantly and apolipoprotein B decreased significantly after 30 g alcohol/d relative to the concentration after the control diet. CONCLUSIONS: Consumption of 15-30 g alcohol/d by postmenopausal women apparently decreases cardiovascular disease risk by improving lipid profiles. Plasma LDL-cholesterol and triacylglycerol concentrations improve after 15 g alcohol/d; plasma HDL cholesterol improves only after 30 g alcohol/d. PMID- 11864869 TI - Ginsenoside variability in American ginseng samples. PMID- 11864871 TI - Are we on a voyage to the unknown? PMID- 11864872 TI - Nystagmus surgery. PMID- 11864874 TI - The overseas ophthalmologist in Britain. PMID- 11864876 TI - Survey of blindness and visual impairment in Bioko, Equatorial Guinea. AB - AIMS: This study was designed to estimate the prevalence of blindness and its main aetiologies in Bioko, an onchocerciasis endemic zone of Equatorial Guinea. METHODS: Random sampling was applied, proportionally to population distribution into urban or rural areas. All the subjects underwent a basic eye examination by trained nurses. In the presence of any ocular affection or a visual acuity of less than 0.3, the subject was visited by the ophthalmologist. This visit included direct and indirect ophthalmoscopy, anterior segment examination with a slit lamp, and intraocular pressure recording. Blindness and visual impairment were defined by using the WHO criteria. RESULTS: 3218 subjects were screened, with a mean age of 34.1 (SD 21.6) years, ranging from 1 month to 102 years (median 34 years). The overall prevalence of blindness was 3.2% (95% CI: 2.6% to 3.9%). Unilateral blindness was present in 4.2%. Visual impairment was diagnosed in 200 patients (6.8%). More than 20% of the acuities inferior to 0.7 improved when explored with a pinhole. The main causes of blindness were cataracts (61.3%); macular affection (25.3%), optic atrophy (16%), and glaucoma (13.3%). Ocular onchocerciasis was detected in 12 cases (0.4%). CONCLUSION: Ocular onchocerciasis was very uncommon in an area of high endemicity. Results also pointed at the lack of basic ophthalmologist eye care and optician resources in the island. PMID- 11864877 TI - Risk factors for graft infection in India: a case-control study. AB - AIM: To study the demographic, clinical, and microbiological profile and the risk factors for graft infection following penetrating keratoplasty. METHODS: 50 eyes of 50 consecutive patients with graft infection after an optical penetrating keratoplasty were included as cases; 50 eyes of 50 patients with no graft infection were included as controls. The main variables evaluated in this study included the clinical and microbiological profile, sociodemographic status, suture related problems, persistent epithelial defects, and ocular surface disorders. RESULTS: Cultures were positive in 43 (86%) eyes and Staphylococcus epidermidis (67.4%) was the most common organism isolated. Infection could be resolved with treatment in 37 (74%) eyes. In eight (16%) eyes the graft melted and a repeat penetrating keratoplasty had to be performed. Only 6% of the cases could achieve a best corrected visual acuity of 6/18 or better after resolution of the infection. In multivariate logistic regression analysis persistent epithelial defect (OR (95% CI): 3.0 (1.17 to 8.33)), suture related problems (OR (95% CI): 3.6 (1.39 to 9.25)), and ocular surface disorders (OR (95% CI): 2.4 (0.93 to 6.03)) were found to be statistically significant risk factors for graft infection following an optical penetrating keratoplasty. CONCLUSIONS: Staphylococcus epidermidis is the commonest organism responsible for post keratoplasty microbial keratitis. Persistent epithelial defects, suture related problems, and ocular surface disorders are the major risk factors predisposing to graft infection. PMID- 11864878 TI - Modified Anderson procedure for correcting abnormal mixed head position in nystagmus. AB - BACKGROUND/AIM: Treatment of nystagmus is controversial mainly in cases where it is combined with abnormal head position. This study was carried out to demonstrate that patients with abnormal head position in all three axes associated with nystagmus show improvement in the torsional and vertical components if only horizontal factors are addressed by surgical weakening of the horizontal muscles. METHODS: 21 patients with horizontal nystagmus and abnormal head position were studied. All had an abnormal head position in all three axes with a predominant head turn. In all cases a modified Anderson procedure was performed---that is, 2 mm retroequatorial recessions of the horizontal yoke rectus muscles responsible for the blockage position, plus corrective surgery for strabismus when needed. RESULTS: The three components of the abnormal head position were improved with surgery of horizontal yoke rectus muscles only (p=0.001). CONCLUSIONS: Large recessions of the horizontal yoke rectus muscles in nystagmus with blockage position, when the head turn predominates over the vertical and torsional components, are effective in diminishing the abnormal head position on all three axes. PMID- 11864879 TI - Phototherapeutic keratectomy re-treatment for recurrent corneal erosion syndrome. AB - AIMS: Phototherapeutic keratectomy with an excimer laser is commonly used to treat recurrent corneal erosion syndrome. The aim of this study was to determine the success of a repeat phototherapeutic keratectomy for patients with persistent macroerosions following initial treatment with phototherapeutic keratectomy. METHODS: Eight patients who failed primary phototherapeutic keratectomy for recurrent corneal erosion syndrome were treated with repeat phototherapeutic keratectomy. All patients were treated with a superficial therapeutic ablation profile with a Visx, Nidek, or Summit 193 nm excimer laser. All patients were treated for both their primary treatment and re-treatment by the same surgeon. Retrospective analysis of case records of all patients requiring re-treatment was supplemented with a telephone interview. RESULTS: Over a 5 year period (October 1995 to October 2000) 76 eyes were treated for recurrent erosion syndrome with phototherapeutic keratectomy. All patients had documented macroerosions and had failed previous treatment with a lubricant at night. Eight eyes (11%) continued to have macroerosions after this initial treatment; all opted for re-treatment with phototherapeutic keratectomy. Following re-treatment none reported symptoms consistent with a macroerosion. Six of eight patients (75%) are now symptom free; 2/8 (25%) have an occasional foreign body sensation relieved by lubricants. Follow up ranged from 9-60 months with a mean of 25.5 months. CONCLUSION: Re treatment with phototherapeutic keratectomy appears to be successful for patients with macroerosions complicating recurrent corneal erosion syndrome who have failed conservative management with ocular lubricants and a primary phototherapeutic keratectomy. PMID- 11864880 TI - Corneal electrolysis for recurrence of corneal stromal dystrophy after keratoplasty. AB - AIMS: To evaluate corneal electrolysis as a treatment for recurrent diffuse corneal opacities at the host-graft interface of the stroma or at the subepithelial region in two types of granular corneal dystrophy (GCD). METHODS: Recurrence developed at the host-graft interface of the stroma after lamellar keratoplasty in a patient with Avellino corneal dystrophy (ACD). At surgery, the deep aspect of the graft in this patient was partially separated from host tissue to expose the deposits, with one third of the host-graft junction left intact. The graft was everted, and electrolysis was applied directly to remove the deposits attached to both surfaces of the host and the graft. Then the graft was returned to its place and sutured. In two patients with homozygous ACD and one patient with the superficial variant of GCD, diffuse subepithelial opacities developed following penetrating keratoplasty. Electrolysis was applied directly to the corneal surface. RESULTS: Deposits at the host-graft interface of the stroma and in the subepithelial region disappeared following treatment, and vision recovered in all patients. CONCLUSIONS: This method is a simple, easy, and inexpensive way to remove deposits that recur after lamellar or penetrating keratoplasty. PMID- 11864881 TI - Biotypes and serotypes of Haemophilus influenzae ocular isolates. AB - AIM: To determine which subtypes of Haemophilus influenzae are most commonly associated with ocular disease, and whether the site of ocular H influenzae infection is correlated with specific subtypes of the organism. METHODS: The biotypes and serotypes of ocular H influenzae isolates collected at the Francis I Proctor Foundation between March 1989 and January 2000 were examined. A total of 62 ocular isolates were retrieved from frozen storage and plated on chocolate agar. Biotypes were assigned based upon the ability of the isolates to produce indole, urease, and ornithine decarboxylase. Capsular subtypes a-f were determined by slide agglutination using commercially available subtype specific antisera. Identified biotypes and serotypes were then analysed with regard to site of infection. RESULTS: Patient age ranged from 1 to 92 years with a median age of 45 years. 38 (61%) of the isolates were biotype II, 23 (37%) were biotype III, and one (2%) was biotype VII. All of the isolates were non-encapsulated and thus serologically non-typable. H influenzae biotype II was found in 28 of 48 (58%) conjunctivitis cases, five of eight (63%) keratitis cases, and two of two (100%) endophthalmitis cases. Biotype III was found in 20 of 48 (42%) conjunctivitis cases, two of eight (25%) keratitis cases, and a single case of dacryocystitis. Biotype VII was associated with one of eight (13%) keratitis cases. CONCLUSION: Most ocular H influenzae isolates appear to be serologically non-typable strains from biotypes II and III, less virulent subtypes that frequently colonise the nasopharynx. In addition, the site of ocular H influenzae infections appears to be largely independent of species subtype. PMID- 11864883 TI - Corkscrew retinal vessels in neurofibromatosis type 1: report of 12 cases. AB - AIM: To describe a distinctive spectrum of retinal microvascular abnormalities in 12 patients with neurofibromatosis type 1 (NF-1). METHODS: This is an observational prospective study of the ocular fundus evaluated by direct ophthalmoscopy with or without fluorescein angiography, to investigate retinal microvascular abnormalities in 32 patients with NF-1 and in 30 control subjects. The evaluation included a complete general and neurological physical examination and in some cases computed tomography, magnetic resonance imaging with gadolinium DTPA, or both. RESULTS: The occurrence of a distinctive spectrum of retinal microvascular abnormalities is described in 12 patients with NF-1 (37.5%). At the lower end of the spectrum, present in 10 patients, the anomaly consisted of minuscule second or third order tortuous venules, which were called "corkscrew retinal vessels." These were usually isolated but in a few cases multiple. They flow towards the superior or inferior temporal veins. They had a length of one to two disc diameters. They ended either in a minute tuft or vanished on the retinal surface. The upper end of the spectrum was seen in only two patients. One of them had an exceptionally large venous anastomosis on the nasal retina and the other had an arteriovenous malformation extending over one retinal quadrant. None of the patients in the control group had such retinal microvascular abnormalities. CONCLUSION: The "corkscrew" retinal vessels described in this report constitute a broad spectrum of microvascular markers in NF-1 patients. PMID- 11864882 TI - Prediction of visual outcome after retinal detachment surgery using the Lotmar visometer. AB - AIM: To evaluate whether an achromatic interferometer, the Lotmar visometer, is useful in predicting postoperative visual outcome in patients with primary rhegmatogenous retinal detachment (RD) involving the macula. METHODS: This prospective study included 40 eyes of 40 non-consecutive patients with macula-off RD. The eyes were phakic or pseudophakic, had a clear optical media, and had a measurable potential vision on preoperative visometric examination. Preoperative variables included Snellen visual acuity, duration of macular detachment, extent of RD, and visometric potential acuity. Reattachment surgery consisted of radial scleral buckling in 33 patients, circumferential scleral buckling and encircling in seven patients, and subretinal fluid drainage in 10 patients. Retinal breaks were treated with cryotherapy or laser photocoagulation. Patients were followed up for at least 6 months after uncomplicated surgery. Best corrected visual acuity measured at any time during follow up was correlated with the preoperative variables. RESULTS: Preoperative visual acuity was less than 20/200 in 37 (93%) of 40 patients. Potential visual acuity of 20/200 or better was measured using the Lotmar visometer in 37 patients (93%). Postoperative visual acuity was correlated significantly with duration of macular detachment (r=0.55; p<0.001), and extent of RD approached statistical significance (r=0.31; p=0.05). There was a higher correlation between postoperative visual acuity and the visometric measurements (r=0.61; p<0.001). CONCLUSIONS: The Lotmar visometer may be a valuable method to estimate visual outcome after uncomplicated scleral buckling surgery in patients with RD involving the macula. PMID- 11864884 TI - Axonal loss from acute optic neuropathy documented by scanning laser polarimetry. AB - BACKGROUND/AIMS: Retinal nerve fibre layer analysis by scanning laser polarimetry has been shown to facilitate diagnosis of glaucoma while its role in glaucoma follow up is still unclear. A major difficulty is the slow reduction of retinal nerve fibre layer thickness in glaucomatous optic neuropathy. Eyes of patients were studied after acute retrobulbar optic nerve lesion in order to evaluate the usefulness of scanning laser polarimetry in documenting retinal nerve fibre layer loss over time. METHODS: Five patients who suffered severe retrobulbar optic neuropathy have had repeated measurements of the retinal nerve fibre layer using scanning laser polarimetry at various intervals, the first examination being within 1 week of injury. RESULTS: All eyes showed a marked decrease in peripapillary retinal nerve fibre layer thickness, which followed an exponential curve and occurred predominantly within 8 weeks of injury. Compared to a previous study using red-free photographs, scanning laser polarimetry showed retinal nerve fibre layer loss earlier in the course of descending atrophy. CONCLUSION: Scanning laser polarimetry is useful for early detection and documentation of retinal nerve fibre layer loss following acute injury to the retrobulbar optic nerve. It seems to be a promising tool for follow up of individual glaucoma patients. PMID- 11864885 TI - Use of bovine pericardium as a wrapping material for hydroxyapatite orbital implants. AB - AIM: To present the results of 27 patients who had enucleation for malignant melanoma of the choroid with hydroxyapatite implant wrapped in bovine pericardium. METHOD: A retrospective study was performed on 27 patients, 12 males and 15 females, who had enucleation as a primary treatment for their choroidal melanomas. The patients were followed up at 1 week, 1 month, 3 months, and then every 6 months. A conformer was fitted at 1 week and an artificial eye at 1 month. The average follow up was 1.7 years. RESULTS: No patient had extrusion of the implant. One patient needed repair of the wound, two patients required a lateral tarsal strip, and one patient developed a conjunctival granuloma, which did not need excision. In one patient there was shallowing of the inferior fornix. The cosmetic results and ocular movement were satisfactory in all but one patient. CONCLUSION: Use of bovine pericardium as wrapping material for the hydroxyapatite implants has shown promising results with minimal extrusion rates providing an effective alternative for sclera, eliminating the potential risks of CJD. PMID- 11864887 TI - Diamond burr superficial keratectomy for recurrent corneal erosions. AB - AIMS: To evaluate the efficacy and safety of diamond burr superficial keratectomy in the treatment of recurrent corneal erosions. METHODS: A retrospective review of 54 eyes (47 patients) with recurrent corneal erosions treated with diamond burr superficial keratectomy. Preoperative and postoperative visual acuities and refractions, slit lamp examination findings, and the incidence of recurrent erosion after keratectomy were studied. Specular microscopy was also performed in six patients before and after surgery. RESULTS: 30 eyes had underlying map dot fingerprint anterior basement membrane corneal dystrophy, while 24 eyes did not. Postoperative follow up time ranged from 3 to 53 months (mean 12.3 months). Corneal erosion recurred in three eyes (6%) after diamond burr superficial keratectomy. This procedure improved the best corrected visual acuity from 20/26 to 20/22 by logMAR statistical evaluation (p=0.002) and caused very little change in the refractive spherical equivalent. No endothelial cell loss or changes in morphology were noted on specular microscopy. CONCLUSION: Diamond burr superficial keratectomy appears to be an effective and safe method of treating recurrent erosions and is a good alternative therapy to needle stromal micropuncture, Nd:YAG induced epithelial adhesion, and excimer laser surface ablation. PMID- 11864886 TI - Neutrophil accumulation correlates with type IV collagenase/gelatinase activity in endotoxin induced uveitis. AB - BACKGROUND/AIM: Anterior uveitis is a common inflammatory ocular disease characterised by protein accumulation and leucocyte infiltration in the anterior chamber. The aim of this study was to determine the expression of gelatinases in the aqueous humour (AH) and uvea in an animal model of endotoxin induced uveitis (EIU). METHODS: EIU was established in Lewis rats following an intraperitoneal injection of lipopolysaccharide (LPS). AH and ocular tissue were obtained from control animals and those with EIU over a 1 week time course and the samples analysed immunohistochemically and by gelatin zymography. RESULTS: Matrix metalloproteinase (MMP) 2 and 9 levels were elevated in rat AH over a 1 week time course. MMP-2 and MMP-9 levels peaked at the time of maximum uveal inflammation, before returning to baseline levels as the inflammation subsided. MMP-9 was detected in the latent and functionally active form. Total protein extracted from inflamed rat uveal tissue displayed no significant gelatinolytic modulation throughout the time course of EIU. Anterior chamber neutrophils and ciliary body epithelial cells were the most abundant source of the gelatinases. CONCLUSION: This study has revealed a correlation between infiltrating neutrophils and the presence of elevated gelatinases in EIU. The results suggest that these proteolytically active enzymes may be important mediators of the inflammatory response and contribute to matrix remodelling observed in uveitis. Furthermore, the excess production of MMPs may be a mechanism by which leucocytes, such as neutrophils, gain access to uveal tissue and AH. Therapeutic strategies aimed at reducing MMP activity may be of some benefit in the treatment of uveitis. PMID- 11864889 TI - Value of two mortality assessment techniques for organ cultured corneal endothelium: trypan blue versus TUNEL technique. AB - BACKGROUND/AIM: It is known that trypan blue staining is not a good predictor of loss of corneal endothelial cells (ECs) during organ culture. As it is primarily an indicator of membrane integrity, it would also not be expected to identify ECs undergoing apoptosis. The aim of this study was to determine the ability of the in situ TdT dUTP mediated nick end labelling (TUNEL) technique to detect cell death in the corneal endothelium caused by apoptosis during organ culture, compared with conventional vital staining with trypan blue. METHODS: 31 human corneas were organ cultured at 31C for 3-35 days. Staurosporine was used to induce apoptosis in five control corneas. The endothelium was assessed by trypan blue and by the in situ TUNEL technique. The percentages of trypan and TUNEL positive ECs were compared. Their links with sex, donor age, time from donor death and organ culture, initial and final EC density and cell loss were studied. RESULTS: TUNEL stained ECs were observed in all corneas. TUNEL positive ECs were mostly located either in corneal folds or at the periphery of corneal folds showing central shedding. The mean percentage of cell death at the end of storage, assessed by the trypan blue technique, was 1.47% (SD 2.63, range 0.03 12); assessed by the TUNEL technique it was 12.7% (SD 16.4 range 0.6-65.5). There was a significant correlation between the two techniques (r = 0.7, p<0.001). The percentage of TUNEL stained ECs was correlated negatively with EC density at the end of storage (r = -0.47, p <0.005) and positively with percentage EC loss during storage (r = 0.46, p < 0.05). CONCLUSION: This study demonstrates that organ cultured corneas systematically carry non-viable ECs that are implicated in cell death by apoptosis and go undetected when trypan blue staining is used. Because the in situ TUNEL assay detects earlier events in the cell death process than does trypan blue, it should be used to quantify endothelial viability, especially for experiments with new storage media. PMID- 11864888 TI - Corneal toxicity secondary to inadvertent use of benzalkonium chloride preserved viscoelastic material in cataract surgery. AB - AIMS: To study the long term toxic effects of intraocular benzalkonium chloride (BAC). METHODS: 19 patients exposed to intraocular BAC preserved viscoelastic during cataract surgery in February 1999 developed severe striate keratopathy immediately postoperatively. 16 patients, including two who underwent penetrating keratoplasty, were studied in the period April to June 2000. Ocular symptoms, visual acuity, biomicroscopy, intraocular pressure, dilated funduscopy, specular endothelial microscopy, and corneal pachymetry findings were recorded. The corneal and iris specimens of the two patients who underwent keratoplasty were studied by light, transmission, and scanning electron microscopy. RESULTS: Six males and 10 females, aged 64-98 years, were studied 14-16 months postoperatively. All patients were symptomatic. 12 patients had best corrected visual acuity of 6/12 or better and four patients of between 6/18 and 6/60. Five patients had corneal epithelial oedema and 11 had Descemet's membrane folds. The central corneal thickness, 620 (SD 71) microm, in affected eyes was significantly higher (p<0.005, two tailed paired t test) than that of the contralateral eyes, 563 (SD 48) microm. The endothelial cell density was significantly lower (p<0.0001, two tailed paired t test) in affected eyes: 830 (SD 280) cells/mm2 v 2017 (SD 446) cells/mm2. The mean average cell area was significantly higher in the BAC treated eyes: 1317 (SD 385) microm2 v 521 (SD 132) microm2. There was no significant difference in the coefficient of variation of cell size between the two eyes (p=0.3, two tailed paired t test). Two corneal specimens displayed morphological features of bullous keratopathy and other non-specific abnormalities. Extracellular melanosomes were present in a portion of the iris of one case. CONCLUSION: BAC is toxic to the corneal endothelium when used intraocularly, leading to severe striate keratopathy. This cleared in most cases but left varying degrees of residual stromal thickening in all eyes. If penetrating keratoplasty is required the results are excellent. PMID- 11864890 TI - Angiotensin II and vascular endothelial growth factor in the vitreous fluid of patients with proliferative diabetic retinopathy. AB - AIMS: To investigate the correlation between the level of angiotensin II and vascular endothelial growth factor (VEGF) in the vitreous fluid and the severity of proliferative diabetic retinopathy (PDR). METHODS: During vitreoretinal surgery at the Tokyo Women's Medical University, vitreous fluid samples were obtained from 51 eyes of diabetic patients with PDR, six eyes of diabetic patients without retinopathy, and 16 eyes of non-diabetic patients with ocular disease (controls). The VEGF levels in vitreous fluid and plasma were determined by enzyme linked immunosorbent assay, while angiotensin II levels were measured by radioimmunoassay. RESULTS: The vitreous fluid levels of VEGF and angiotensin II were significantly higher in patients with PDR than in non-diabetic patients or diabetic patients without retinopathy (all p<0.0001). The vitreous fluid level of angiotensin II was significantly correlated with that of VEGF (p<0.0001), and the vitreous concentrations of both VEGF and angiotensin II were significantly higher in patients with active PDR than in those with quiescent PDR (p<0.0001 and p=0.0005, respectively). CONCLUSION: The authors found that both angiotensin II and VEGF levels were significantly higher in the vitreous fluid of patients with PDR than in that of non-diabetic patients or diabetic patients without retinopathy, and that the levels of both angiotensin II and VEGF were elevated in the active stage of PDR. These findings suggest that angiotensin II contributes to the development and progression of PDR in combination with VEGF. PMID- 11864891 TI - Diabetic macular oedema: a comparison of vitreous fluorometry, angiography, and retinopathy. AB - AIM: To evaluate the relation between the quantitative measurement of vitreous fluorescein with fluorescein angiography and retinopathy in diabetic patients with and without clinically significant macular oedema (CSMO). METHODS: In a prospective cross sectional study, passive permeability and active, outward transport of fluorescein across the blood-retinal barrier were quantitated with vitreous fluorometry in 61 eyes from 48 patients with CSMO and 22 fellow eyes without CSMO, after exclusion of eyes with previous macular laser treatment and vitreous liquification. All patients were recruited from the university hospital's outpatient clinic. Retinopathy and fluorescein angiograms were evaluated on 60 degree photographs. RESULTS: The passive permeability in CSMO was significantly correlated with the severity of leakage on fluorescein angiograms (r=0.73), the level of retinopathy (r=0.61), and visual acuity (r=0.45). Significant differences between eyes with CSMO and eyes without CSMO were found for passive permeability (p<0.001), fluorescein leakage (p<0.001), visual acuity (p=0.02), and retinopathy (p=0.002). CONCLUSION: Passive permeability of fluorescein quantitated with vitreous fluorometry was correlated both with semiquantitative fluorescein angiography and retinopathy, and a significant increase in passive permeability was found when comparing eyes with CSMO to eyes without CSMO. No such pattern was found for the active transport indicating that passive and not the outward, active transport is the factor of most importance in the development of CSMO. PMID- 11864892 TI - Polypoidal choroidal vasculopathy treated with macular translocation: clinical pathological correlation. AB - AIMS: To report the histopathology of two specimens of polypoidal choroidal vasculopathy (PCV) obtained from two eyes of Japanese patients. METHODS: Specimens were obtained under direct visualisation during macular translocation surgery with 360 degree retinotomy. The clinical findings were correlated with the light microscopic findings of the two specimens. RESULTS: One specimen from a 77 year old man was the central portion of the lesion that lay under the sensory retina on the retinal pigment epithelium (RPE). The specimen was made up mainly of fibrous tissue with small, thin walled vessels. Indocyanine green angiography after surgery revealed that active leaking polypoidal element remained under the RPE. Another specimen obtained from a 62 year old man was made up of a fibrovascular membrane situated within Bruch's membrane. The part of this specimen inferior to the foveal region included a collection of dilated, thin walled blood vessels without pericytes, surrounded by macrophages that stained positive for CD68. The dilated vessels appeared to be correlated with the orange coloured polyps observed by ophthalmoscopy, the polypoidal structure seen in indocyanine green angiograms, and the pyramidal elevation with intermediate reflectivity by optical coherence tomography. CONCLUSION: Polypoidal structures are located within Bruch's space. They are composed of clusters of dilated, thin walled blood vessels surrounded by macrophages and fibrin material. The positive immunohistochemical staining for vascular endothelial growth factor in the RPE and the vascular endothelial cells suggests that this fibrovascular complex is a subretinal choroidal neovascularisation. PMID- 11864893 TI - A novel mutation of the RP1 gene (Lys778ter) associated with autosomal dominant retinitis pigmentosa. AB - BACKGROUND: Besides the three known genes (RHO, RDS/Peripherin, NRL) involved in autosomal dominant retinitis pigmentosa (adRP), a fourth gene, RP1, has been recently identified. Initial reports suggest that mutations in the RP1 gene are the second most frequent cause of adRP. The clinical findings were described in a family with adRP and a novel mutation in the RP1 gene. METHOD: Index patients from 15 independent families with adRP in which RHO mutations had been excluded in previous examinations were screened for mutations in the RP1 gene by means of direct DNA sequencing. Evaluation of the RP1 phenotype in patients included funduscopy, kinetic perimetry, dark adapted final threshold test, standard electroretinography and, in one case, multifocal electroretinography. RESULTS: One novel nonsense mutation (Lys778ter) in one of these 15 patients was detected. Cosegregation of the mutation with the disease phenotype could be established in the index patient's family. The phenotype comprises variable expression of clinical disease probably including one case of incomplete penetrance, a onset of symptoms beginning in adulthood, and evidence of regionally varying retinal function loss. CONCLUSION: The Lys778ter mutation localises inside the critical region harbouring all mutations described so far. The ophthalmic findings support previous observations that variation of disease expression appears as a typical feature of the RP1 phenotype. PMID- 11864894 TI - Mode of presentation and time to treatment of uveal melanoma in Finland. AB - AIMS: To investigate the current referral pattern and delays in treatment of patients with primary uveal melanoma. METHODS: 184 consecutive Finnish patients with uveal melanoma diagnosed between July 1994 and June 1999 were eligible, and 159 were enrolled (inclusion rate, 86%). Their mean age was 60 years (range 14 87). The dates of visits to dispensing optician, physician, ophthalmologist and ocular oncologist, the presence of symptoms, and reason for consultation were determined by structured telephone interview. Time intervals to treatment planning and treatment were calculated. RESULTS: 139 patients (87%) had symptoms at presentation and 44 patients (28%) had been seen by an ophthalmologist less than 2 years previously. The median height of the tumour was 6 mm (range, 1.0 17.0) and its largest basal diameter 11 mm (range 2.5-22.0) at diagnosis. Melanoma developed from a previously detected presumed naevus in 13 patients (8%). When the first contact was a dispensing optician (15%) the median time to treatment planning was 22 days (range 1-1156). When a physician other than an ophthalmologist (19%) was contacted the delay was 68 days (range 0-1283) and when an ophthalmologist (65%) was seen it was 34 days (range 1-1426). These differences were not significant (p=0.32). The chance of being referred at first visit was 89%. Median time to treatment was not associated with symptoms (p=0.16) and tumour volume (p=0.29), but it was significantly different between patients who were and were not referred at first visit (140 days v 34 days; p<0.001) and between those treated by ruthenium and iodine brachytherapy (59 days v 33 days; p=0.009). CONCLUSIONS: Analysis of delays in management indicates that earlier treatment could be achieved if dilated fundus examinations were performed without exceptions, all suspicious naevi were referred for a second opinion, and if the patients with melanoma were referred to the ocular oncology service concurrently with staging examinations done at the regional hospital. PMID- 11864895 TI - Longitudinal study of trachomatous trichiasis in the Gambia. AB - AIM: Investigation of the natural history of trachomatous trichiasis in the Gambia and of the outcome of self epilation and surgery for the condition. METHODS: A 1 year longitudinal study of 190 subjects with trichiasis was performed. Major trichiasis cases (five lashes or more) were referred for surgery and minor trichiasis cases were advised to epilate. Outcome measures included progression of trichiasis and corneal scarring; attendance for and results of surgery. RESULTS: 34 of 148 (23%, 95% CI 16 to 31) subjects with major trichiasis attended for surgery over the year. Progression from minor to major trichiasis occurred in 18 of 55 subjects (33%, 95% CI 21 to 47). Progression of corneal scarring occurred in 60 of 167 patients (36%, 95% CI 29 to 44). Clinically active trachoma and conjunctival bacterial isolation predicted progression of corneal opacity. Surgery was successful in 39 of 54 (72%) eyes. CONCLUSIONS: Despite the overall decline in trachoma in the Gambia, patients with both minor and major trichiasis remain at risk of developing corneal opacity. Active trachomatous inflammation and additional infection with bacteria may accelerate this process. Antibiotic treatment for trichiasis patients (in addition to surgery) should be investigated. Surgery for minor trichiasis may be indicated. Regular audit of surgical results is necessary with retraining where needed. PMID- 11864896 TI - Evaluation of the validity and reliability of A-scan ultrasound biometry with a single use disposable cover. AB - BACKGROUND: The UK Medical Devices Agency has suggested that ophthalmic practitioners should, where practicable and not compromising clinical outcome, restrict corneal contact devices to single patient use to minimise a remote theoretical risk of transmission of new variant Creutzfeldt-Jakob disease (vCJD). This study reports on a modified technique of ultrasound A-scan biometry that complies with the MDA recommendations. METHODS: The right eyes of 37 consecutive hospital patients had a series of biometry readings taken with a Humphrey 820 A scan instrument with a plane wave transducer use d conventionally and with the addition of a disposable latex cover. RESULTS: Intrasessional repeatability of axial length measurements was similar for conventional readings--mean difference 0.027 mm, 95% confidence intervals (CI) +/- 0.44 mm and those taken with a disposable cover (0.028 mm, CI +/- 0.38). Intersessional repeatability was equivalent with (0.002 mm, CI +.- 0.51) and without a cover (0.03 mm, CI +/- 0.51). Readings with a cover were not significantly different from those without (paired t test; p >0.05), but tended to be greater (mean difference 0.085 mm, CI +/- 0.60). CONCLUSIONS: These findings suggest that corneal contact biometry with a disposable cover is a viable and theoretically safer alternative to the conventional technique. PMID- 11864898 TI - Henoch-Schonlein purpura with bilateral central retinal artery occlusion. PMID- 11864897 TI - The ocular surface toxicity of paraquat. PMID- 11864899 TI - Primary iris pigment epithelial hyperplasia and glaucoma. PMID- 11864900 TI - Unilateral vitreous haemorrhage secondary to caudal epidural injection: a variant of Terson's syndrome. PMID- 11864901 TI - Severe macular pucker after retinal detachment surgery in an infant. PMID- 11864902 TI - Microbial keratitis associated with extended wear of silicone hydrogel contact lenses. PMID- 11864903 TI - Serious corneoscleral complications after pterygium excision with mitomycin C. PMID- 11864904 TI - Extrascleral extension of a choroidal melanoma after argon photocoagulation and transpupillary thermotherapy. PMID- 11864905 TI - Macular infarction after intravitreal amikacin: mounting evidence against amikacin. PMID- 11864906 TI - Spontaneous involution of retinal and intracranial arteriovenous malformation in Bonnet-Dechaume-Blanc syndrome. PMID- 11864907 TI - These eyes are hot... PMID- 11864908 TI - Pcp1p, an Spc110p-related calmodulin target at the centrosome of the fission yeast Schizosaccharomyces pombe. AB - In the budding yeast Saccharomyces cerevisiae, the calmodulin-binding protein Spc110p/Nuf1p facilitates mitotic spindle formation from the fungal centrosome or spindle pole body (SPB). The human Spc110p orthologue kendrin is a centrosomal, calmodulin-binding pericentrin isoform that is specifically overexpressed in carcinoma cells. Here we establish an evolutionary and functional link between Spc110p and kendrin through identification and analysis of similar calmodulin binding proteins in the fission yeast Schizosaccharomyces pombe (Pcp1p, pole target of calmodulin in S. pombe) and the filamentous fungus Aspergillus nidulans. Like Spc110p and kendrin, Pcp1p and the A. nidulans protein contain predicted coiled-coil secondary structure and a COOH-terminal calmodulin-binding region. Green fluorescent protein fusions of Pcp1p localize to the SPB as analyzed by fluorescence and immunoelectron microscopy. Pcp1p overexpression causes chromosome missegregation, multiple mitotic spindle fragments, and multiple abnormal SPB-like structures, a phenotype remarkably similar to that of many human carcinoma lines, which exhibit chromosome and spindle defects, and supernumerary centrosomes. PMID- 11864909 TI - SV40 T antigen and telomerase are required to obtain immortalized human adult bone cells without loss of the differentiated phenotype. AB - In most human primary bone cells, SV40 T-antigen expression was able to expand life span for a few passages before cells undergo growth arrest, described as crisis. In this study, telomerase activity was reconstituted in human osteoblast precursors (hPOB cells) and marrow stromal cells (Saka cells) transformed with the SV40 T antigen. Bone cells with telomerase activity were able to bypass crisis and show unlimited life span. Despite chromosomal aberrations observed in hPOB-tert cells, these immortalized precursors were able to differentiate into osteoblasts like precrisis hPOB cells. Saka-tert cells enhanced the formation of human osteoclast-like cells in a similar manner as Saka cells. These results demonstrate that reconstitution of telomerase activity in transformed SV40 T antigen human osteoblast precursors or marrow stromal cells leads to the generation of immortalized cells with a preserved phenotype. PMID- 11864910 TI - Interaction between the hematopoietic Ets transcription factor Spi-B and the coactivator CREB-binding protein associated with negative cross-talk with c-Myb. AB - We have previously shown that the hematopoietic Ets transcription factor PU.1 interacts with the transcriptional coactivator CREB-binding protein (CBP). In this study, we further investigated whether Spi-B, another hematopoietic Ets transcription factor, also interacts with CBP. Direct physical interaction of Spi B with CBP was demonstrated by glutathione S-transferase binding assay. Analysis using several deletion mutants of Spi-B and CBP revealed that the NH2-terminal region including the activation domain of Spi-B interacted with the region spanning amino acid residues 1283-1915 of CBP in vitro. The interaction of Spi-B with CBP was also observed in vivo. CBP potentiated Spi-B-mediated transcription of the reporter gene driven by the multimerized PU.1/Spi-B binding sites. This transcriptional activation by Spi-B and CBP was inhibited by expression of c-Myb, and the transcriptional activation by c-Myb and CBP was inhibited by expression of Spi-B, suggesting competition for CBP between these two transcription factors. Our results suggest that CBP acts as a transcriptional coactivator of Spi-B and mediates synergistic or antagonistic interactions between other transcription factors. PMID- 11864911 TI - Carcinogen-induced S-phase arrest is Chk1 mediated and caffeine sensitive. AB - We have investigated the mechanism of S-phase arrest elicited by the carcinogen benzo(a)pyrene dihydrodiol epoxide (BPDE) in p53-deficient cells. Inhibition of DNA synthesis after BPDE treatment was rapid and dose dependent (approximately 50% inhibition after 2 h with 50 nM BPDE). Cells treated with low doses (50-100 nM) of BPDE resumed DNA synthesis after a delay of approximately 4-8 h, whereas cells that received high doses of BPDE (600 nM) failed to recover from S-phase arrest. The checkpoint kinase Chk1 (but not Chk2) was phosphorylated after treatment with low doses of BPDE. High concentrations of BPDE elicited phosphorylation of both Chk1 and Chk2. Adenovirus-mediated expression of "dominant-negative" Chk1 (but not dominant-negative Chk2) and the Chk1 inhibitor UCN-01 abrogated the S-phase delay elicited by low doses of BPDE. Consistent with a role for the caffeine-sensitive ATM or ATR protein kinase in low-dose BPDE induced S-phase arrest, both Chk1 phosphorylation and S-phase arrest were abrogated by caffeine. However, low doses of BPDE elicited Chk1 phosphorylation and S-phase arrest in AT cells (from ataxia telangiectasia patients), demonstrating that ATM is dispensable for S-phase checkpoint responses to this genotoxin. BPDE-induced Chk1 phosphorylation and S-phase arrest were abrogated by caffeine treatment in AT cells, suggesting that a caffeine-sensitive kinase other than ATM is an important mediator of responses to BPDE-adducted DNA. Overall, our data demonstrate the existence of a caffeine-sensitive, Chk1-mediated, S-phase checkpoint that is operational in response to BPDE. PMID- 11864912 TI - Insulin-like growth factor-I-induced migration of melanoma cells is mediated by interleukin-8 induction. AB - Successive events of growth factor-induced autocrine and paracrine activation promote tumor growth and metastasis. Insulin-like growth factor-I (IGF-I) stimulates melanoma cells to grow, survive, and migrate. Interleukin-8 (IL-8) is produced by melanoma cells and has been correlated with melanoma metastasis, but the biological functions of this cytokine have not been elucidated. We show here that IGF-I-induced migration of melanoma cells could be inhibited by neutralizing antibody against IL-8. IGF-I overexpression induced IL-8 production in melanoma cells, especially in biologically early melanomas by accelerating its transcription rate via activation of mitogen-activated protein kinase pathway. IGF-I treatment phosphorylated c-Jun and stimulated the binding of AP-1 but not NF-kappaB to the IL-8 promoter. These data identify IL-8 as a new target of IGF-I in melanoma and suggest that some of the biological functions of IGF-I are mediated by IL-8. PMID- 11864914 TI - Looking for the pony in the HERS data. Heart and Estrogen/progestin Replacement Study. PMID- 11864916 TI - Coronary magnetic resonance angiography in adolescents and young adults with kawasaki disease. AB - BACKGROUND: In patients with Kawasaki disease, serial evaluation of the distribution and size of coronary artery aneurysms (CAA) is necessary for risk stratification and therapeutic management. Although transthoracic echocardiography is often sufficient for this purpose initially, visualization of the coronary arteries becomes progressively more difficult as children grow. We sought to prospectively compare coronary magnetic resonance angiography (MRA) and x-ray coronary angiography findings in patients with CAA caused by Kawasaki disease. METHODS AND RESULTS: Six subjects (age 10 to 25 years) with known CAA from Kawasaki disease underwent coronary MRA using a free-breathing T2-prepared 3D bright blood segmented k-space gradient echo sequence with navigator gating and tracking. All patients underwent x-ray coronary angiography within a median of 75 days (range, 1 to 359 days) of coronary MRA. There was complete agreement between MRA and x-ray angiography in the detection of CAA (n=11), coronary artery stenoses (n=2), and coronary occlusions (n=2). Excellent agreement was found between the 2 techniques for detection of CAA maximal diameter (mean difference=0.4 +/- 0.6 mm) and length (mean difference=1.4 +/- 1.6 mm). The 2 methods showed very similar results for proximal coronary artery diameter (mean difference=0.2 +/- 0.5 mm) and CAA distance from the ostia (mean difference=0.1 +/- 1.5 mm). CONCLUSION: Free-breathing 3D coronary MRA accurately defines CAA in patients with Kawasaki disease. This technique may provide a non-invasive alternative when transthoracic echocardiography image quality is insufficient, thereby reducing the need for serial x-ray coronary angiography in this patient group. PMID- 11864915 TI - Targeting phospholamban by gene transfer in human heart failure. AB - BACKGROUND: Myocardial cells from failing human hearts are characterized by abnormal calcium handling, a negative force-frequency relationship, and decreased sarcoplasmic reticulum Ca2+ ATPase (SERCA2a) activity. In this study, we tested whether contractile function can be improved by decreasing the inhibitory effects of phospholamban on SERCA2a with adenoviral gene transfer of antisense phospholamban (asPL). METHODS AND RESULTS: Myocardial cells isolated from 9 patients with end-stage heart failure and 18 donor nonfailing hearts were infected with adenoviruses encoding for either the antisense of phospholamban (Ad.asPL), the SERCA2a gene (Ad.SERCA2a), or the reporter genes beta galactosidase and green fluorescent protein (Ad.betagal-GFP). Adenoviral gene transfer with Ad.asPL decreased phospholamban expression over 48 hours, increasing the velocity of both contraction and relaxation. Compared with cardiomyocytes infected with Ad.asPL (n=13), human myocytes infected with Ad.betagal-GFP (n=8) had enhanced contraction velocity (20.3 +/- 3.9% versus 8.7 +/- 2.6% shortening/second; P<0.01) and relaxation velocity (26.0 +/- 6.2% versus 8.6 +/- 4.3% shortening/second; P<0.01). The improvement in contraction and relaxation velocities was comparable to cardiomyocytes infected with Ad.SERCA2a. Failing human cardiomyocytes had decreased contraction and Ca2+ release with increasing frequency (0.1 to 2 Hz). Phospholamban ablation restored the frequency response in the failing cardiomyocytes to normal; increasing frequency resulted in enhanced sarcoplasmic reticulum Ca2+ release and contraction. CONCLUSION: These results show that gene transfer of asPL can improve the contractile function in failing human myocardium. Targeting phospholamban may provide therapeutic benefits in human heart failure. PMID- 11864917 TI - Platelets activated by collagen through immunoreceptor tyrosine-based activation motif play pivotal role in initiation and generation of neointimal hyperplasia after vascular injury. AB - BACKGROUND: Platelet adhesion on components of the extracellular matrix and platelet activation by those components are crucial for the arrest of posttraumatic bleeding, but they can also harm tissue by occluding diseased vessels. Recent studies have shown that the activation of platelets by collagen is mediated through the same pathway used by immune receptors, with an immunoreceptor tyrosine-based activation motif on the Fc receptor gamma chain (FcRgamma) playing a pivotal role. METHODS AND RESULTS: We examined the role of collagen-stimulated platelets in the development of injury-induced neointimal formation by using mice deficient in FcRgamma. The left femoral arteries of 8- to 12-week-old FcRgamma-deficient mice (n=16) and C57BL/6 (wild-type) mice (n=16) were injured by a straight spring wire (0.35-mm diameter). Segments of the injured and uninjured femoral arteries were excised at 7 days and 28 days after the vascular injury. Arterial segments were examined by immunohistochemistry and electron microscopy. Two hours after injury, electron microscopy showed marked decreases in platelet adhesion and neutrophil attachment to the vascular wall surface in FcRgamma-knockout mice compared with wild-type mice. At 7 days after injury, staining with anti-neutrophil antibody showed fewer neutrophils in FcRgamma-knockout mice than in wild-type mice. Computer-aided morphometry performed to measure the neointimal area, intima/media ratio, and stenotic area at 28 days after injury showed a significantly smaller ratio and area in FcRgamma knockout mice than in wild-type mice (for neointimal area, 16 635 +/- 1406 versus 31 483 +/- 2309 microm2, respectively; for intima/media ratio, 1.25 +/- 0.40 versus 2.68 +/- 0.04, respectively; and for stenotic area, 26.8 +/- 2.1% versus 49.3 +/- 4.1%, respectively). CONCLUSIONS: These results demonstrate that FcRgamma may play important roles in the initiation and generation of neointimal hyperplasia after balloon injury through the activation of platelets by collagen. PMID- 11864918 TI - Subgroup interactions in the Heart and Estrogen/Progestin Replacement Study: lessons learned. AB - BACKGROUND: The Heart and Estrogen/Progestin Replacement Study (HERS) showed no overall benefit of postmenopausal hormone treatment in women with coronary heart disease (CHD). We analyzed the HERS data to determine whether there were specific subgroups of women who responded differently to treatment, either during the first year or in the overall study. METHODS AND RESULTS: In the search for significant treatment interactions, we analyzed a total of 86 subgroups defined by baseline characteristics. These included demographics and lifestyle factors, laboratory and physical examination variables, medical history and symptoms by self-report, medication use, and prior CHD history by chart review. We examined within-subgroup treatment effects for baseline variables that significantly interacted with treatment assignment. Under the null hypothesis, 4 (5%) of the 86 interactions would be expected to be nominally significant (P<0.05) by chance alone at each time point. Six of the interaction values were P<0.05 at 1 year, and 3 were P<0.05 at trial completion. The findings are discussed in the context of known mechanisms of action and prior scientific knowledge. Use of digitalis and history of myocardial infarction emerged as 2 possible modifiers of the effect of hormone therapy during the first year, and lipoprotein(a) emerged as a possible modifier during the overall study. CONCLUSIONS: Extensive post hoc analyses did not identify any subgroup of HERS participants in which postmenopausal hormone treatment was clearly beneficial or harmful, but several possibilities emerged for testing in future trials. PMID- 11864919 TI - Detection of lipid pool, thin fibrous cap, and inflammatory cells in human aortic atherosclerotic plaques by near-infrared spectroscopy. AB - BACKGROUND: A method is needed to identify nonstenotic, lipid-rich coronary plaques that are likely to cause acute coronary events. Near-infrared (NIR) spectroscopy can provide information on the chemical composition of tissue. We tested the hypothesis that NIR spectroscopy can identify plaque composition and features associated with plaque vulnerability in human aortic atherosclerotic plaques obtained at the time of autopsy. METHODS AND RESULTS: A total of 199 samples from 5 human aortic specimens were analyzed by NIR spectroscopy. Features of plaque vulnerability were defined by histology as presence of lipid pool, thin fibrous cap (<65 microm by ocular micrometry), and inflammatory cell infiltration. An InfraAlyzer 500 spectrophotometer was used. Spectral absorbance values were obtained as log (1/R) data from 1100 to 2200 nm at 10-nm intervals. Principal component regression was used for analysis. An algorithm was constructed with 50% of the samples used as a reference set; blinded predictions of plaque composition were then performed on the remaining samples. NIR spectroscopy sensitivity and specificity for histological features of plaque vulnerability were 90% and 93% for lipid pool, 77% and 93% for thin cap, and 84% and 89% for inflammatory cells, respectively. CONCLUSIONS: NIR spectroscopy can identify plaque composition and features associated with plaque vulnerability in postmortem human aortic specimens. These results support efforts to develop an NIR spectroscopy catheter system to detect vulnerable coronary plaques in living patients. PMID- 11864920 TI - Effect of Abciximab on prothrombin activation and thrombin generation in acute coronary syndromes without ST-segment elevation: Global Utilization of Strategies to Open Occluded Coronary Arteries Trial IV in Acute Coronary Syndromes (GUSTO IV ACS) Italian Hematologic Substudy. AB - BACKGROUND: Abciximab is very effective in reducing major cardiac events in patients undergoing interventional procedures. Its antithrombotic effect is primarily attributable to the blocking of platelet glycoprotein IIb/IIIa receptors, but recent evidence suggests that it may have a direct antithrombin effect. No data are available concerning the effect of abciximab on the in vivo markers of prothrombin activation and thrombin generation in patients with acute coronary syndromes without ST elevation. METHODS AND RESULTS: We measured the plasma levels of prothrombin fragment 1+2 (a marker of prothrombin activation) and the thrombin/antithrombin complex (a marker of thrombin generation) in 167 patients with acute coronary syndromes without ST elevation enrolled in the GUSTO IV ACS trial who were randomized to receive abciximab for 24 hours (52 patients), abciximab for 48 hours (59 patients), or placebo (56 patients) in addition to heparin. Blood samples were obtained at baseline (before any treatment), after 24 and 48 hours (before study drug discontinuation), and 1 month later. There was a significant increase in the plasma levels of prothrombin fragment 1+2 after 48 hours and after 1 month in all 3 groups, placebo (P=0.0001), 24-hour abciximab (P=0.0002), and 48-hour abciximab (P=0.0001). The plasma thrombin/antithrombin complex levels were similar in the 3 groups at all time points and did not change during the study drug infusions. CONCLUSIONS: Abciximab does not decrease prothrombin activation and thrombin generation in patients with acute coronary syndromes without ST elevation not undergoing interventional procedures. PMID- 11864921 TI - Increased nitric oxide bioavailability in endothelial cells contributes to the pleiotropic effect of cerivastatin. AB - BACKGROUND: Although statins preserve endothelial function by reducing serum cholesterol levels, it has been suggested they may also stimulate nitric oxide (NO) synthase in endothelium with concurrent increase in superoxide (O2-) generation, leading to impairment of NO activity. Therefore, measurements of biologically active NO and O2- in endothelium after exposure to the HMG-CoA reductase inhibitor cerivastatin were undertaken to evaluate its potential effect on NO biological activity. METHODS AND RESULTS: Highly sensitive electrochemical NO and O2- microsensors were placed near the surface of a single human umbilical vein endothelial cell, and the kinetics of NO and O2- release were recorded in vitro. Cerivastatin demonstrated a time-dependent effect on NO release in endothelial cells. The initial release (approximately the first 3 minutes) was concentration-dependent (0.01 to 10 micromol/L) and was similar to that observed for typical NO synthase agonists calcium ionophore or acetylcholine. Cerivastatin stimulated NO release at a favorable rate and scavenged O2-, which led to the preservation of the active concentration of NO. The sustained effect (after approximately 6 hours) of cerivastatin on endothelium was associated with an approximately 35% increase in NO release as compared with the initial effect. In contrast to the initial effect, the sustained effect of cerivastatin was shown at concentrations approximately 100-fold lower and was dependent on inhibition of endothelial HMG-CoA reductase. CONCLUSIONS: These data provide direct evidence to prove that in the presence of cerivastatin, the NOS system in endothelium operates with high efficiency toward increasing NO activity by activation of NO release and by concurrent inactivation of O2-. PMID- 11864922 TI - Relationship between coronary artery remodeling and plaque vulnerability. AB - BACKGROUND: In vivo studies with intravascular ultrasound have shown that complex plaque anatomy and plaque rupture are more frequent in the presence of marked outward remodeling. A large lipid core and a high macrophage count are recognized histological markers for plaque vulnerability. The link between plaque vulnerability in terms of these markers and remodeling in coronary arteries has not been explored. METHODS AND RESULTS: In 88 male subjects who died suddenly with coronary artery disease, 108 plaques were studied. The percent remodeling was calculated. Lesions with remodeling > or = 0% were considered to have positive remodeling, and those in which remodeling was < 0% were considered to have negative remodeling. Percent lipid core and macrophage count at the plaque were assessed. Of 108 plaque sites, 64 (59.2%) had undergone no remodeling or positive remodeling, and 44 (40.7%) had negative remodeling (vessel shrinkage). Lesions with positive remodeling, compared with lesions with vessel shrinkage, had a larger lipid core (percent mean lipid core was 39.0 +/- 21.0% versus 22.3 +/- 23.1%, respectively; P < 0.0001) and a higher macrophage count (mean macrophage count was 15.6 +/- 12.3 versus 8.9 +/- 11.6, respectively; P = 0.005). CONCLUSIONS: We have shown that coronary artery plaques with positive remodeling have a higher lipid content and macrophage count, both markers of plaque vulnerability. These results may explain why plaque rupture is often apparent at sites with only modest luminal stenoses (but marked positive remodeling). PMID- 11864923 TI - Remodeling of left ventricular hypertrophy in elite athletes after long-term deconditioning. AB - BACKGROUND: The clinical significance and long-term consequences of left ventricular (LV) hypertrophy associated with intensive athletic conditioning remain unresolved. METHODS AND RESULTS: We prospectively evaluated 40 elite male athletes who had shown marked LV cavity enlargement of > or = 60 mm, wall thickness of > or = 13 mm, or both in a longitudinal fashion with serial echocardiograms, initially at peak training (age 24 +/- 4 years) and subsequently after a long-term deconditioning period (1 to 13 years; mean, 5.6 +/- 3.8). After detraining, LV cavity dimension decreased by 7% (61.2 +/- 2.9 to 57.2 +/- 3.1 mm; P<0.001), maximum wall thickness by 15% (12.0 +/- 1.3 to 10.1 +/- 0.8 mm; P<0.001), and mass normalized to height by 28% (194 +/- 25 to 140 +/- 21 g/m; P<0.001). However, individual subject analysis showed persistent substantial cavity dilatation (> or = 60 mm) in 9 athletes (22%); in contrast, wall thickness returned to normal in each athlete. Multiple regression analysis demonstrated that approximately 50% of the incomplete reduction in cavity dimension was explained by increased body weight and recreational physical activity performed during the follow-up period. No athlete had developed cardiac symptoms, impaired exercise performance, or evidence of LV dysfunction. CONCLUSIONS: LV remodeling was evident after long-term detraining, with significant reduction in cavity size and normalization of wall thickness. Resolution of cavity enlargement was, however, incomplete in most cases, and substantial chamber dilatation persisted in >20% of athletes. The possibility that this residual LV hypertrophy, apparently part of the athlete's heart syndrome, may have future long-term clinical implications in some individuals cannot be excluded with certainty. PMID- 11864924 TI - Peroxisome proliferator--activated receptor alpha gene regulates left ventricular growth in response to exercise and hypertension. AB - BACKGROUND: Left ventricular hypertrophy (LVH) occurs as an adaptive response to a physiological (such as exercise) or pathological (valvular disease, hypertension, or obesity) increase in cardiac work. The molecular mechanisms regulating the LVH response are poorly understood. However, inherited defects in fatty acid oxidation are known to cause severe early-onset cardiac hypertrophy. Peroxisome proliferator--activated receptor alpha (PPARalpha) regulates genes responsible for myocardial fatty acid oxidation and is downregulated during cardiac hypertrophy, concomitant with the switch from fatty acid to glucose utilization. METHODS AND RESULTS: The role of PPARalpha in left ventricular growth was investigated in 144 young male British Army recruits undergoing a 10 week physical training program and in 1148 men and women participating in the echocardiographic substudy of the Third Monitoring Trends and Determinants in Cardiovascular Disease (MONICA) Augsburg study. A G/C polymorphism in intron 7 of the PPARalpha gene significantly influenced left ventricular (LV) growth in response to exercise (P=0.009). LV mass increased by 6.7 +/- 1.5 g in G allele homozygotes but was significantly greater in heterozygotes for the C allele (11.8 +/- 1.9 g) and in CC homozygotes (19.4 +/- 4.2 g). Likewise, C allele homozygotes had significantly higher LV mass, which was greater still in hypertensive subjects, and a higher prevalence of LVH in the Third MONICA Augsburg study. CONCLUSIONS: We demonstrate that variation in the PPARalpha gene influences human left ventricular growth in response to exercise and hypertension, indicating that maladaptive cardiac substrate utilization can play a causative role in the pathogenesis of LVH. PMID- 11864925 TI - Aging attenuates the vestibulosympathetic reflex in humans. AB - BACKGROUND: The vestibular system contributes to sympathetic activation by engagement of the otolith organs. However, there is a significant loss of vestibular function with aging. Therefore, the purpose of the present study was to determine if young and older individuals differ in their cardiovascular and sympathetic responses to otolithic stimulation (ie, head-down rotation, HDR). We hypothesized that responses to otolithic stimulation would be attenuated in older adults because of morphological and physiological alterations that occur in the vestibular system with aging. METHODS AND RESULTS: Arterial blood pressure, heart rate, muscle sympathetic nerve activity (MSNA), and head rotation were measured during HDR in 11 young (26 +/- 1 years) and 11 older (64 +/- 1 years) subjects in the prone posture. Five older subjects performed head rotation (chin to chest) in the lateral decubitus position, which simulates HDR but does not alter afferent inputs from the vestibular system. MSNA responses to HDR were significantly attenuated in older as compared with young subjects (P<0.01). MSNA increased in the older subjects by only 12 +/- 5% as compared with 85 +/- 16% in the young. Furthermore, HDR elicited significant reductions in mean arterial blood pressure in older (Delta-6 +/- 1 mm Hg; P<0.01) but not young subjects (Delta1 +/- 1 mm Hg). In contrast to HDR, head rotation performed in the lateral decubitus position did not elicit hypotension. MSNA responses to baroreceptor unloading and the cold pressor test were not different between the age groups. CONCLUSIONS: These data indicate that aging attenuates the vestibulosympathetic reflex in humans and may contribute to the increased prevalence of orthostatic hypotension with age. PMID- 11864926 TI - HMG-CoA reductase inhibitors suppress intracellular calcium mobilization and membrane current induced by lysophosphatidylcholine in endothelial cells. AB - BACKGROUND: Lysophosphatidylcholine (LPC) is known to increase intracellular Ca2+ concentration ([Ca2+]i) in endothelial cells. This study was conducted to investigate the effects of HMG-CoA reductase inhibitors (statins) on the increase in [Ca2+]i and membrane current induced by LPC. METHODS AND RESULTS: [Ca2+]i was determined in cultured human aortic endothelial cells by fura-2 assay, and membrane current was measured by whole-cell patch clamp. The [Ca2+]i increase induced by LPC was abolished by inhibitors of phospholipase C (PLC). Statins markedly decreased the [Ca2+]i increase caused by LPC. This suppressive effect was quickly reversed by geranylgeranylpyrophosphate (GGPP) and was mimicked by inhibitors of Rho and Rho kinase. LPC induced the translocation of the GTP-bound active form of RhoA into membranes within 1 minute as determined by a pull-down assay and reduced the levels of RhoA in the cytoplasm, indicating that LPC quickly increases the GTP/GDP ratio of RhoA and induces membrane translocation. Statins prevented the GTP/GDP exchange of RhoA and its membrane translocation from the cytoplasm caused by LPC, and these effects of statins were reversed by GGPP. The responses of RhoA activation to statins and GGPP concurred with their effects on Ca2+ mobilization. LPC also induced a nonselective cation current after a lag. Statins prolonged the lag and decreased the current amplitude, and GGPP abolished the inhibitory effect on the current. CONCLUSIONS: LPC induced Ca2+ mobilization and membrane current via a Rho activation-dependent PLC pathway in endothelial cells, and statins blocked these effects by preventing the GGPP dependent lipid modification of Rho. The present study implicates Rho in LPC stimulation of Ca2+ movement. PMID- 11864927 TI - Protection of human endothelial cells from oxidative stress: role of Ras-ERK1/2 signaling. AB - BACKGROUND: Reactive oxygen species play a critical role in inducing apoptosis. The small GTPase p21 Ras and the ERK1/2 MAPK have been proposed as key regulators of the signaling cascade triggered by oxidative stress (H2O2). Harvey-Ras (Ha Ras) and Kirsten-Ras (Ki-Ras) isoforms are so far functionally indistinguishable, because they activate the same downstream effectors, including ERK1/2. Moreover, ERK1/2 signaling has been involved in both protection and induction of apoptosis. METHODS AND RESULTS: Human umbilical vein endothelial cells (HUVECs) were subjected to H2O2, and apoptosis was detected by fluorescence-activated cell sorting analysis, fluorescence microscopy, and caspase-3 activation. Transfection of Ha-Ras and Ki-Ras genes in HUVECs was performed to evaluate the response to H2O2. We have found that, whereas Ha-Ras decreases tolerance to oxidative stress, Ki-Ras has a potent antiapoptotic activity. Both effects are mediated by ERK1/2. Tolerance to H2O2 is encoded by a unique stretch of lysines at the COOH terminus of the Ki-Ras, lacking in Ha-Ras, and it is relatively independent of the farnesylated anchor. Inhibition of p21 Ras signaling by farnesylation inhibitors increased the resistance to apoptosis in Ha-Ras-expressing cells. CONCLUSIONS: These findings explain the opposite effects of ERK1/2 stimulation on apoptosis found in different cell types and suggest that local activation of ERK1/2 signaling may account for the opposing response to oxidative stress by Ha-Ras or Ki-Ras-expressing cells. Modulation of cell reactivity to oxidative stress by p21 Ras points to the specific and predictive effects of Ras inhibitors in vivo as potential therapeutic drugs in disorders produced by increase of reactive oxygen species inside the cells. PMID- 11864928 TI - Different effects of carvedilol, metoprolol, and propranolol on left ventricular remodeling after coronary stenosis or after permanent coronary occlusion in rats. AB - BACKGROUND: Although carvedilol attenuates left ventricular (LV) remodeling in coronary occlusion-reperfusion, it is not known whether it attenuates ischemic LV remodeling because of coronary stenosis (CS) or permanent coronary occlusion (CO). METHODS AND RESULTS: We administered a vehicle, carvedilol, propranolol (2, 10, and 30 mg/kg per day, each), metoprolol (6, 30, and 90 mg/kg per day), or bunazosin (0.2 and 1 mg/kg per day), orally for 12 weeks to a total of 608 rats with CS or CO. In these groups and the sham (n=40), we assessed LV function by echocardiography, CS severity, myocardial blood flow and coronary flow reserve, serum ascorbyl free radical, and vitamin C. Both CS and CO increased LV end diastolic and end-systolic diameters and decreased ejection fraction. The 4 agents failed to attenuate LV remodeling caused by CO. In contrast, the 3 beta blockers attenuated (P<0.01) or tended to attenuate the increase in LV end diastolic diameters caused by CS. With similar blood pressure and heart rate lowering by 3 beta-blockers, carvedilol additionally attenuated the increase in end-systolic diameters and improved ejection fraction. The CS reduced myocardial blood flow and coronary flow reserve, which was reversed by carvedilol without modifying the CS severity. Among the 4 agents, only carvedilol decreased ascorbyl free radical and increased vitamin C. CONCLUSIONS: The effects of beta blockade on ischemic cardiac dysfunction seem to depend on the different properties of the beta-blockers and the doses used. Among the beta-blockers tested, carvedilol provided potent cardioprotection for compromised ischemic but viable myocardium rather than for infarcted myocardium. PMID- 11864929 TI - Postischemic recovery of contractile function is impaired in SOD2(+/-) but not SOD1(+/-) mouse hearts. AB - BACKGROUND: Reactive oxygen species (ROS) contribute to myocardial stunning. Superoxide dismutase (SOD) is a major defense mechanism against ROS. The purpose of this study was to evaluate the contributions of cytosolic (SOD1) and mitochondrial (SOD2) isoforms to protect against myocardial stunning. METHODS AND RESULTS: Isolated hearts from wild-type, heterozygous (+/-) SOD1 and SOD2 knockout mice received 30 minutes of ischemia followed by 60 minutes of reperfusion. After 60 minutes of reperfusion, the heart rate multiplied by the developed pressure (HRxDP) in the wild-type and SOD1(+/-) hearts recovered to 92 +/- 9 and 85 +/- 7 of preischemic baseline values, respectively (P=NS). In contrast, the HRxDP was significantly lower (63 +/- 7%) in the SOD2(+/-) hearts compared with the wild-type hearts. Western blot analysis and enzymatic activity of tissue lysates confirmed reduction of activities of specific SOD isoforms without compensatory increase in the other isoform in the knockout animals studied. CONCLUSIONS: Postischemic functional recovery is more sensitive to a partial deficiency of SOD2 than a partial deficiency of SOD1. Therefore, modulation of the mitochondrial SOD isoform is a critical determinant in the tolerance of the heart to oxidative stress. PMID- 11864930 TI - Perfusion versus function: the ischemic cascade in demand ischemia: implications of single-vessel versus multivessel stenosis. AB - BACKGROUND: We hypothesized that during demand ischemia, abnormal perfusion will precede abnormal function, the spatial extent of perfusion abnormality will be greater than that of functional abnormality, and the spatiotemporal disparity between abnormal perfusion and abnormal function will be more marked in the presence of single-vessel stenosis (SVS) versus multivessel stenosis (MVS). METHODS AND RESULTS: Nine dogs each underwent either SVS or MVS placement. These noncritical stenoses were classified as mild, moderate, or severe on the basis of the transstenotic pressure gradient (10 to 14, 15 to 20, or >20 mm Hg). Dobutamine was infused starting at 10 and reaching 40 microg/kg(-1) x min(-1). Wall thickening (WT) and myocardial perfusion (myocardial contrast echocardiography) were assessed at each stage. Resting perfusion and function were normal in all dogs. In SVS, abnormal perfusion (delayed rate of microbubble replenishment) was seen at the lowest dose of dobutamine irrespective of the stenosis severity, whereas WT abnormality was seen only at high doses of dobutamine and was influenced by the stenosis severity. The spatial extent of abnormal perfusion exceeded that of WT abnormality at all but the highest dobutamine dose. This spatiotemporal discordance between abnormal perfusion and function was significantly less in MVS, where it was possible to identify separate regions with abnormal function at lower doses of dobutamine. CONCLUSIONS: These data support the occurrence of the ischemic cascade during demand ischemia. They also explain the higher sensitivity of abnormal perfusion compared with abnormal function for the detection of coronary stenosis as well as the higher sensitivity of dobutamine echocardiography for MVS compared with SVS. PMID- 11864931 TI - Progression and regression of atherosclerotic lesions: monitoring with serial noninvasive magnetic resonance imaging. AB - BACKGROUND: Modification or stabilization of atherosclerotic plaques has been proposed as the mechanism responsible for the beneficial clinical effect of lipid lowering therapies. An imaging modality able to quantify atherosclerotic plaque composition could potentially allow not only the identification of these vulnerable atherosclerotic lesions, but also monitoring of the effects of therapeutic interventions on plaque composition. Our aim was to monitor changes in burden and composition of atherosclerotic plaques in a rabbit model of complex atherosclerosis using serial noninvasive magnetic resonance imaging (MRI). METHODS AND RESULTS: Aortic atherosclerotic lesions were induced in rabbits and the animals randomized to continue an atherogenic diet (atherosclerosis progression) or resume normal chow (atherosclerosis regression) for 6 months. MRI of the aorta was performed at 3 time points in each rabbit, as follows: baseline, after atherosclerosis induction (9 months old), and after atherosclerosis regression or progression (15 months old). Histopathologic correlation with MRI was performed. There was a significant (P<0.0001) reduction in atherosclerotic burden in the regression group, and an increase in the progression group. There was a significant (P<0.001) reduction in the lipidic component of plaques in the regression group, and an increase in the progression group. A small, nonsignificant increase in the fibrotic plaque components was noted in the regression group, but a significant decrease in the fibrotic composition of lesions in the progression group. A significant correlation (P<0.001) was found between MRI and histopathology for atherosclerotic burden and plaque composition. CONCLUSIONS: These findings indicate that serial noninvasive MRI can monitor changes in atherosclerotic plaque composition under conditions of atherosclerotic progression and regression. PMID- 11864932 TI - Maximizing the natriuretic peptide system in experimental heart failure: subcutaneous brain natriuretic peptide and acute vasopeptidase inhibition. AB - BACKGROUND: A hallmark of congestive heart failure (CHF) is the elevation of the cardiac natriuretic peptides (NPs), which have natriuretic, renin-inhibiting, vasodilating, and lusitropic properties. We have reported that chronic subcutaneous (SQ) administration of brain natriuretic peptide (BNP) in experimental CHF improves cardiorenal function. Vasopeptidase inhibitors (VPIs) are single molecules that simultaneously inhibit both neutral endopeptidase 24.1 (NEP) and ACE. We hypothesized that acute VPI administration would potentiate the cardiorenal actions of SQ BNP in experimental CHF. METHODS AND RESULTS: We determined the cardiorenal and humoral responses to acute VPI alone with omapatrilat (OMA) (1 micromol/kg IV bolus) (n=6), acute low-dose SQ BNP (5 microg/kg) alone (n=5), acute VPI plus low-dose SQ BNP (n=5), and acute high-dose SQ BNP (25 microg/kg) alone in 4 groups of anesthetized dogs with experimental CHF produced by ventricular pacing for 10 days. Plasma BNP was greater with VPI+low-dose SQ BNP compared with VPI alone or low-dose SQ BNP alone and was similar to high-dose SQ BNP alone. Urinary BNP excretion was greatest with VPI+SQ BNP. Urinary sodium excretion was also highest with VPI+SQ BNP, with the greatest increase in glomerular filtration rate. VPI+SQ BNP resulted in a greater increase in cardiac output and reduction in cardiac filling pressures as compared with low dose SQ BNP, high-dose SQ BNP, or VPI alone. CONCLUSIONS: This study reports that acute VPI potentiates the cardiorenal actions of SQ BNP in experimental CHF. This study advances the concept that protein therapy with BNP together with vasopeptide inhibition represents a novel therapeutic strategy in CHF to maximize the beneficial properties of the natriuretic peptide system. PMID- 11864933 TI - Direct thrombin inhibitors in acute coronary syndromes: present and future. PMID- 11864934 TI - Heat shock proteins, inflammation, and cardiovascular disease. PMID- 11864935 TI - Images in cardiovascular Medicine. Magnetic resonance contrast enhancement of iatrogenic septal myocardial infarction in hypertrophic cardiomyopathy. PMID- 11864936 TI - Images in cardiovascular medicine. Left ventricular apical pseudoaneurysm: echocardiographic and intraoperative findings. PMID- 11864937 TI - Synergism among flavonoids in inhibiting platelet aggregation and H2O2 production. PMID- 11864938 TI - Serum triglyceride concentration and coronary heart disease. PMID- 11864939 TI - Paradoxical clinical value of another smoker's paradox. PMID- 11864941 TI - Second verse of HERS same as the first--no clear benefit or harm for cardiovascular disease. PMID- 11864940 TI - Large artery stiffness and baroreflex function. PMID- 11864942 TI - The in vivo-in vitro paradox in pneumococcal respiratory tract infections. PMID- 11864943 TI - Role of aminoglycosides in the treatment of bacterial endocarditis. AB - Aminoglycosides are often used in combination with glycopeptides and beta-lactams for the treatment of bacterial endocarditis caused by alpha-haemolytic streptococci, staphylococci and enterococci. Recommendations on dosing and duration of aminoglycoside therapy vary; we detail the laboratory and clinical evidence base for the treatment regimens advised. A number of deficiencies in the published guidelines are highlighted, as is the limited evidence base upon which many of the regimens are recommended. With the increasing use of outpatient treatment of severe infections and once daily administration of aminoglycosides the opportunity should be taken for larger, multicentre clinical trials to evaluate formally both old and new regimens. PMID- 11864944 TI - Pharmacodynamic considerations in the treatment of moderate to severe pseudomonal infections with cefepime. AB - Pseudomonas aeruginosa is one of the more common and clinically difficult-to treat causes of hospital-acquired infections. Cefepime is a broad-spectrum cephalosporin with potent in vitro activity against Gram-positive cocci, enteric Gram-negative bacilli and Pseudomonas aeruginosa. Cephalosporins exhibit time dependent bactericidal activity and lack prolonged post-antibiotic effects against Enterobacteriaceae and P. aeruginosa. In non-clinical models of infection against Enterobacteriaceae and P. aeruginosa, antibacterial effects are observed when serum levels are above the MIC for as little as 35% of the dosing interval and are maximized when levels exceed the MIC for 60-70% of the dosing interval. Based on the MIC distribution for P. aeruginosa and pharmacokinetic data obtained from patients with serious bacterial infections (including pneumonia and sepsis), time above MIC targets can be met in infected patients following 2 g doses of cefepime administered every 12 h. An understanding of the integration of target patient population pharmacokinetics and the MIC distribution is crucial for selecting effective dosage regimens, especially in the setting of empirical therapy. Moreover, sufficient clinical outcome data in infected patients exist and support these pharmacodynamic conclusions. PMID- 11864945 TI - In vitro and in vivo antibacterial activity of T-3912, a novel non-fluorinated topical quinolone. AB - The in vitro and in vivo activity of T-3912, a novel non-fluorinated topical quinolone, was compared with that of nadifloxacin, ofloxacin, levofloxacin, clindamycin, erythromycin and gentamicin. The in vitro activity of T-3912 against methicillin-susceptible Staphylococcus aureus, ofloxacin-resistant and methicillin-resistant S. aureus, Staphylococcus epidermidis, ofloxacin-resistant S. epidermidis, penicillin-resistant Streptococcus pneumoniae and Propionibacterium acnes was four-fold to 16 000-fold greater than that of other agents at the MIC90 for the clinical isolates. The activity of T-3912 was not influenced by grlA mutation in S. aureus, and the degree of MIC increase of T 3912 for grlA-gyrA double and triple mutants was lowest among the quinolones tested (nadifloxacin, levofloxacin and ofloxacin). The inhibitory activity of T 3912 was compared with other quinolones for DNA gyrase and topoisomerase IV of S. aureus SA113. T-3912 showed the greatest inhibitory activity for both enzymes among the quinolones tested. The isolation frequency of spontaneous mutants resistant to T-3912 was < 1.7 x 10(-9) and < 2.0 x 10(-9) for S. aureus SA113 and P. acnes JCM 6425, respectively. Furthermore, resistance to T-3912 could not be clearly detected in the 28th transfer by the serial passage method. T-3912 exhibited more potent bactericidal activity against S. aureus and P. acnes than nadifloxacin and clindamycin in a short time period. T-3912 in a 1% gel formulation showed good therapeutic activity against a burn infection model caused by S. aureus SA113, P. acnes JCM6425 and multidrug-resistant S. aureus F 2161. These results indicate that T-3912 is potentially a useful quinolone for the treatment of skin and soft-tissue infections and that its potent bactericidal activity might be able to shorten the treatment period. PMID- 11864946 TI - In vitro bactericidal activity of daptomycin against staphylococci. AB - MICs and minimum bactericidal concentrations (MBCs) of daptomycin, vancomycin, linezolid and quinupristin-dalfopristin (Q-D) were determined for 108 staphylococcal isolates. All strains were susceptible (MICs) to daptomycin (< or =2.0 mg/L) and Q-D (< or =1.0 mg/L). All but three isolates were susceptible to vancomycin (< or =4.0 mg/L) and all but one methicillin-resistant Staphylococcus aureus strain were susceptible to linezolid (< or =4.0 mg/L). Q-D had the lowest geometric mean MIC (0.29 mg/L) and daptomycin had the lowest geometric mean MBC (0.57 mg/L). Time-kill tests were performed on 25 isolates. Bactericidal activity (>99.9% kill) was observed with daptomycin at 2 mg/L and at 2 x MBC for 92% of strains tested. In comparison, the bactericidal rates for the other drugs at breakpoint concentrations and at 2 x MBC were 72% and 70% for vancomycin, 46% and 60% for Q-D, and 7% and 14% for linezolid. Of the four drugs tested, daptomycin was bactericidal against the most strains and had the most rapid cidal activity. PMID- 11864947 TI - Activity of clinafloxacin, compared with six other quinolones, against Acinetobacter baumannii clinical isolates. AB - The in vitro activity of clinafloxacin was studied in comparison with ciprofloxacin, levofloxacin, moxifloxacin, nalidixic acid, sparfloxacin and trovafloxacin against Acinetobacter baumannii clinical isolates. Clinafloxacin showed a MIC(90) of 4 mg/L, whereas the remaining quinolones showed a MIC(90) equal to or higher than 16 mg/L. MIC(50) determination in the presence of reserpine resulted in a two-fold decrease, except for trovafloxacin, which decreased four-fold, and for moxifloxacin and nalidixic acid, which did not change. The effect of reserpine was most pronounced among strains with a low level of resistance to quinolones. The MIC of clinafloxacin for strains with no mutation in either gyrA or parC genes ranged from 0.008 to 0.25 mg/L. In strains with a single mutation at amino acid codon Ser83 of the gyrA gene, the MIC of clinafloxacin ranged from 0.12 to 1 mg/L, whereas strains with a double mutation, one in the gyrA gene and another in the parC gene, showed a range of MIC of clinafloxacin from 1 to 8 mg/L. Therefore, clinafloxacin shows good activity against strains carrying a single mutation in the gyrA gene, and hence a second mutation is required for the microorganism to express resistance. PMID- 11864948 TI - Antibiotic resistance among clinical isolates of Acinetobacter in the UK, and in vitro evaluation of tigecycline (GAR-936). AB - A survey was conducted of the antimicrobial susceptibilities of 595 Acinetobacter spp. isolated from routine clinical specimens in 54 sentinel laboratories throughout the UK during 2000. Isolates of the Acinetobacter baumannii complex (genomic groups 2, 3 and 13TU; n = 443) were distinguished from other genomic groups (n = 152) by PCR fingerprinting of tDNA spacer regions. MICs of amikacin, cefotaxime, ceftazidime, ciprofloxacin, colistin, gentamicin, imipenem, meropenem, minocycline, piperacillin, piperacillin/tazobactam, rifampicin, sulbactam and tetracycline were determined on IsoSensitest agar and interpreted, wherever possible, using BSAC breakpoints. Tigecycline (GAR-936), a new glycylcycline, was also tested. Resistance to cephalosporins, aminoglycosides and ciprofloxacin was widespread, but carbapenems, colistin, sulbactam, minocycline and tigecycline were each active against >80% of the isolates. Isolates of A. baumannii were more often resistant to cefotaxime, ceftazidime, piperacillin, piperacillin/tazobactam, ciprofloxacin, gentamicin and tetracyclines than those belonging to other genomic groups, but were less often resistant to colistin; no significant differences between genomic groups were noted in the susceptibilities to amikacin, carbapenems, rifampicin or sulbactam. The relative activities of the tetracyclines were minocycline > tigecycline > tetracycline. Thirteen carbapenem resistant isolates (MICs > or =8 mg/L; 2.2%) were received from six centres; four centres sent single isolates; one sent three and one sent six. An allele of bla(IMP) was detected in one of these isolates, but the other 12 isolates either had carbapenemase-independent resistance, or undetectable carbapenemase activity combined with other resistance mechanisms. In conclusion, carbapenems, colistin and minocycline retained greatest activity against the Acinetobacter isolates collected. Tigecycline was less active than minocycline, but both agents overcame most tetracycline resistance. PMID- 11864950 TI - Antibiotic resistance of faecal enterococci in poultry, poultry farmers and poultry slaughterers. AB - The prevalence of resistance in enterococci to antibiotics, commonly used for therapy in poultry or as antimicrobial growth promoters (AMGPs), was determined in faecal samples of two chicken populations: broilers in which antibiotic and AMGP use is common and laying-hens with a low antibiotic usage. In addition faecal samples were examined from three human populations: broiler farmers, laying-hen farmers and poultry slaughterers. MICs of an extended panel of antibiotics for a randomly chosen gentamicin- or vancomycin-resistant enterococcal isolate from each faecal specimen were also determined. The prevalence of resistance for all antibiotics tested was higher in broilers than in laying-hens. Resistance in faecal enterococci of broiler farmers was for nearly all antibiotics higher than those observed in laying-hen farmers and poultry slaughterers. The overall resistance in broilers was correlated with the resistance in broiler farmers and in poultry slaughterers. No correlation between the results obtained in the laying-hens with any of the other populations was found. The 27 gentamicin-resistant isolates all showed high-level resistance to gentamicin and two of these isolates, both Enterococcus faecium, were resistant to all antibiotics tested, except vancomycin. The 73 vancomycin-resistant enterococci (VRE) isolated from the five populations belonged to four different species and in all isolates the vanA gene cluster was detected by blot hybridization. The pulsed-field gel electrophoresis (PFGE) patterns of these vancomycin-resistant enterococci were quite heterogeneous, but Enterococcus hirae isolates with the same or a closely related PFGE pattern were isolated at two farms from the broiler farmer and from broilers. Molecular characterization of vanA-containing transposons of these isolates showed that similar transposon types, predominantly found in poultry, were present. Moreover, similar vanA elements were not only found in isolates with the same PFGE pattern but also in other VRE isolated from both humans and chickens. The results of this study suggest transmission of resistance in enterococci from animals to man. For VRE this might be clonal transmission of animal strains, but transposon transfer seems to occur more commonly. PMID- 11864949 TI - Prevalence of resistance to macrolide, lincosamide and streptogramin antibiotics in Gram-positive cocci isolated in a Korean hospital. AB - To investigate the prevalence of resistance to macrolide, lincosamide and streptogramin (MLS) antibiotics in Gram-positive cocci isolated in a Korean hospital, we tested the antibiotic susceptibility of 1097 clinical isolates of Staphylococcus aureus, coagulase-negative staphylococci (CNS) and enterococci to the macrolides erythromycin, clarithromycin, azithromycin and josamycin, the lincosamide clindamycin and the streptogramin pristinamycin. These three groups of organisms were mostly resistant to macrolides and lincosamide, but were commonly susceptible to pristinamycin. The resistance phenotypes of erythromycin resistant isolates were determined by the double-disc test with erythromycin and clindamycin, which showed that most exhibited constitutive MLS resistance. In order to determine the prevalence of the resistance genotypes and the resistance mechanisms, the presence of the erm(A), erm(B), erm(C) and mef genes in the erythromycin-resistant isolates was identified by PCR analysis. The resistance was due mainly to the presence of erm(A) in S. aureus (82.5%), erm(B) in enterococci (55%) and erm(C) in CNS (47.2%). PMID- 11864951 TI - Experimental study of the efficacy of vancomycin, rifampicin and dexamethasone in the therapy of pneumococcal meningitis. AB - The object of the study was to assess the efficacy of rifampicin and the combination of rifampicin plus vancomycin in a rabbit model of experimental penicillin-resistant pneumococcal meningitis. We also studied the effect of concomitant dexamethasone on the CSF antibiotic levels and inflammatory parameters. The rabbit model of pneumococcal meningitis was used. Groups of eight rabbits were inoculated with 106 cfu/mL of a cephalosporin-resistant pneumococcal strain (MIC of cefotaxime/ceftriaxone 2 mg/L). Eighteen hours later they were treated with rifampicin 15 mg/kg/day, vancomycin 30 mg/kg/day or both plus minus dexamethasone (0.25 mg/kg/day) for 48 h. Serial CSF samples were withdrawn to carry out bacterial counts, antibiotic concentration and inflammatory parameters. Rifampicin and vancomycin promoted a reduction of >3 log cfu/mL at 6 and 24 h, and cfu were below the level of detection at 48 h. Combination therapy with vancomycin plus rifampicin was not synergic but it had similar efficacy to either antibiotic alone and it was able to reduce bacterial concentration below the level of detection at 48 h. Concomitant use of dexamethasone decreased vancomycin levels when it was used alone (P< 0.05), but not when it was used in combination with rifampicin. Rifampicin alone at 15 mg/kg/day produced a rapid bactericidal effect in this model of penicillin-resistant pneumococcal meningitis. The combination of vancomycin and rifampicin, although not synergic, proved to be equally effective. Using this combination in the clinical setting may allow rifampicin administration without emergence of resistance, and possibly concomitant dexamethasone administration without significant interference with CSF vancomycin levels. PMID- 11864952 TI - Heterogeneous mechanisms of azole resistance in Candida albicans clinical isolates from an HIV-infected patient on continuous fluconazole therapy for oropharyngeal candidosis. AB - Molecular mechanisms of azole resistance in Candida albicans include alterations in the target enzyme and increased efflux of drug, but the impact of specific treatment regimens on resistance has not been established. A patient with advanced AIDS was enrolled in a longitudinal study to receive continuous oral fluconazole (FLU) 200 mg/day for the treatment of oropharyngeal candidosis (OPC). Oral cultures were obtained at time of enrollment, during episodes of OPC and quarterly for surveillance. The patient had five symptomatic relapses on continuous FLU during 43 months. All OPC episodes were successfully treated with increasing doses of FLU although increased FLU MICs were detected for C. albicans isolates with progression of time. DNA-typing techniques demonstrated that resistance developed in a persistent strain of C. albicans. Both FLU-resistant and isogenic isolates with reduced susceptibility were detected in the same clinical samples through multiple episodes. Analysis of molecular mechanisms of resistance revealed overexpression of MDR and CDR genes encoding efflux pumps (but not ERG11) in isolates with decreased FLU susceptibility. In addition, the presence of the G464S amino acid substitution in their lanosterol demethylase, affecting its affinity for FLU, was also detected. However, other isogenic, but FLU-susceptible isolates recovered from the same samples did not harbour the mutation, indicating microevolution of yeast populations within the oral cavity. In this patient, the continuous antifungal pressure exerted by FLU resulted in development of resistance of multifactorial nature. Despite their clonal origin, different subpopulations of C. albicans demonstrated distinct resistance mechanisms, including concomitant presence and absence of functional point mutations in ERG11 genes. PMID- 11864953 TI - Liposomal amphotericin B and granulocyte colony-stimulating factor therapy in a murine model of invasive infection by Scedosporium prolificans. AB - We established a reproducible lethal disseminated infection by the opportunistic fungus Scedosporium prolificans in an immunosuppressed murine model. We compared the effectiveness of the combined administration of liposomal amphotericin B (LAMB) and granulocyte colony-stimulating factor (G-CSF) with that of either agent alone and with that of amphotericin B deoxycholate (AMB). LAMB + G-CSF and LAMB treatments improved survival significantly with respect to the untreated control. The mean survival times of these three groups were 13.2, 9.1 and 7.9 days, respectively. Culture results in terms of colony counts for samples of deep organs were lower in mice treated with the combined therapy, although differences were not significant. Combined LAMB + G-CSF therapy could be a promising approach for the treatment of disseminated infections of S. prolificans, although further studies are required to determine the most appropriate doses. PMID- 11864954 TI - Potential of rifamides to inhibit TNF-induced NF-kappaB activation. AB - Rifamides are important components in the treatment of tuberculosis. However, it is well documented that these drugs can have immunosuppressive activity, a property of these drugs that is particularly relevant to AIDS patients. In this study, we have shown that a number of rifamide analogues have the potential to block tumour necrosis factor (TNF)- or phorbol myristate acetate-induced NF kappaB activation. As TNF is important in the host defence against tuberculosis, suppression of this activity may provide a potential mechanism of rifamide immunosuppressive activity. PMID- 11864955 TI - BAL 9141, a new broad-spectrum pyrrolidinone cephalosporin: activity against clinically significant anaerobes in comparison with 10 other antimicrobials. AB - The in vitro potency of BAL 9141, a new pyrrolidinone cephalosporin, was tested against non-duplicate strains of anaerobic bacteria. The MIC(50) was 1 mg/L against Actinomyces species, Clostridium species, Gram-positive anaerobic cocci, Porphyromonas species, Fusobacterium species, Lactobacillus species, Prevotella species and Veillonella species. The MIC(50) was 16 mg/L for Bacteroides fragilis and other Bacteroides species. BAL 9141 was not active against cefoxitin resistant Bacteroides fragilis. PMID- 11864956 TI - In vitro activity of rifampicin against Helicobacter pylori isolated from children and adults. AB - Antibiotic resistance in Helicobacter pylori has been a growing problem with current treatment regimens. However, second-line therapy for cases with eradication failure remains to be established. The aim of this study was to evaluate (in vitro) the potential use of rifampicin for H. pylori eradication. The susceptibility of 52 H. pylori strains isolated from children and adults to amoxicillin, clarithromycin, metronidazole and rifampicin was determined. Forty H. pylori isolates susceptible to these antibiotics were investigated for in vitro inducement of resistance. All isolates were susceptible to rifampicin. None of the H. pylori isolates tested acquired rifampicin resistance during prolonged exposure in in vitro inducement experiments. It is suggested that rifampicin has stable activity against H. pylori. PMID- 11864957 TI - What about antibiotic resistance in Neisseria lactamica? AB - The in vitro activity of penicillin, ampicillin, cefotaxime, ceftriaxone, rifampicin and ciprofloxacin against 286 Neisseria lactamica isolates was determined by agar dilution and the category of susceptibility was analysed in accordance with the criteria used for Neisseria meningitidis. All isolates were considered to have intermediate susceptibility to penicillin. A total of 1.7% of the isolates were resistant to ampicillin but all were susceptible to cefotaxime and ceftriaxone. Rifampicin MICs ranged between 0.12 and 2 mg/L. Six isolates (2.1%) showed decreased susceptibility to ciprofloxacin. PMID- 11864958 TI - Use of a rapid mismatch PCR method to detect gyrA and parC mutations in ciprofloxacin-resistant clinical isolates of Escherichia coli. AB - Four amino acid substitutions, two in GyrA and two in ParC subunits of DNA gyrase and topoisomerase IV, respectively, are commonly responsible for fluoroquinolone resistance in Escherichia coli. In this study, an economical and time-efficient mismatch amplification mutation assay (MAMA) PCR was developed to detect mutations in the chromosomal gyrA and parC genes causing these substitutions. One hundred and twenty-one clinical E. coli isolates were tested by this assay, and the results confirmed that accumulation of amino acid alterations in GyrA and ParC correlates closely with stepwise increases in the MIC of ciprofloxacin. PMID- 11864959 TI - Non-PmrA-mediated multidrug resistance in Streptococcus pneumoniae. AB - The PmrA multidrug transporter protein gene was inactivated in Streptococcus pneumoniae strains CP1000 (wild-type) and EBR (mutant with enhanced active multidrug efflux). While the resistance to fluoroquinolones and ethidium bromide shown by EBR was reduced to the wild-type level, neither the susceptibility to reserpine in the presence of ethidium bromide and selected fluoroquinolones, nor the ability to produce ethidium bromide-resistant mutants was eliminated in the CP1000 pmrA mutant, indicating the presence of an additional multidrug export protein(s). PMID- 11864960 TI - The ACT-1 plasmid-encoded AmpC beta-lactamase is inducible: detection in a complex beta-lactamase background. AB - The purpose of this study was to identify the genetic organization and inducibility of bla(ACT-1) in a clinical isolate of Klebsiella pneumoniae possessing at least five different beta-lactamases. The genetic organization of the bla(ACT-1)/ampR region is identical to those of inducible chromosomal ampC genes. RNA analysis using primer extension demonstrated a five-fold increase in bla(ACT-1) transcript production on exposure to cefoxitin. These findings are significant because induction was detected in a complicated beta-lactamase background. In addition, this report is the first to describe an inducible plasmid-encoded AmpC beta-lactamase of Enterobacter cloacae origin. PMID- 11864961 TI - A nosocomial outbreak of Pseudomonas aeruginosa isolates expressing the extended spectrum beta-lactamase GES-2 in South Africa. AB - Eight Pseudomonas aeruginosa clinical strains that produce the clavulanic-acid inhibited beta-lactamase GES-2 were isolated from patients of a South African hospital from March to July 2000. They were clonally related and each harboured a 150 kb conjugative plasmid carrying a class 1 integron containing a gene cassette encoding GES-2, followed by those for beta-lactamase OXA-5 and an aminoglycoside modifying AAC(3)I-like enzyme. Hence, incidences of infection, several fatal, due to bacteria displaying clavulanate-inhibited resistance to extended-spectrum cephalosporins and reduced susceptibility to imipenem in Pretoria Academic Hospital, South Africa, can be explained, at least in part, by the spread of P. aeruginosa expressing the GES-2 beta-lactamase. PMID- 11864962 TI - Influence of cefazolin prophylaxis and hospitalization on the prevalence of antibiotic-resistant bacteria in the faecal flora. AB - Faecal samples from 180 patients on admission and 108 patients at discharge were analysed for the prevalence and degree of resistance of Escherichia coli and enterococci. Discharged patients had received 24 h cefazolin prophylaxis and had a mean hospital stay of 10 +/- 5 days. The prevalence of resistance of E. coli to amoxicillin (P < 0.05), cefazolin (P < 0.05) and oxytetracycline was higher for patients at discharge (41, 12 and 35%) than for patients at admission (28, 2 and 27%). The prevalence at discharge was higher after short-term (< or =7 days) than after long-term (>7 days) hospitalization, being significant for amoxicillin only (P < 0.05). No significant changes were observed for the enterococci. PMID- 11864963 TI - Identification of a cassette-borne dfrA7-like gene that shows a 97 bp extension at the 3'-end of the reading frame. PMID- 11864964 TI - In vitro activity of faropenem against respiratory pathogens. PMID- 11864965 TI - In vitro activity of linezolid against Gram-positive isolates causing infection in continuous ambulatory peritoneal dialysis patients. PMID- 11864966 TI - In vitro activity of new ketolides against macrolide-susceptible and -resistant Staphylococcus aureus isolates with defined resistance gene status. PMID- 11864967 TI - Antimicrobial resistance of invasive Streptococcus pneumoniae in Slovenia from 1997 to 2000. PMID- 11864968 TI - Purification, cloning, and characterization of Nek8, a novel NIMA-related kinase, and its candidate substrate Bicd2. AB - We describe the isolation, cloning, and characterization of human Nek8, a new mammalian NIMA-related kinase, and its candidate substrate Bicd2. Nek8 was isolated as a beta-casein kinase activity in rabbit lung and has an N-terminal catalytic domain homologous to the Nek family of protein kinases. Nek8 also contains a central domain with homology to RCC1, a guanine nucleotide exchange factor for the GTPase Ran, and a C-terminal coiled-coil domain. Like Nek2, Nek8 prefers beta-casein over other exogenous substrates, has shared biochemical requirements for kinase activity, and is capable of autophosphorylation and oligomerization. Nek8 activity is not cell cycle regulated, but like Nek3, levels are consistently higher in G(0)-arrested cells. During the purification of Nek8 a second protein co-chromatographed with Nek8 activity. This protein, Bicd2, is a human homolog of the Drosophila protein Bicaudal D, a coiled-coil protein. Bicd2 is phosphorylated by Nek8 in vitro, and the endogenous proteins associate in vivo. Bicd2 localizes to cytoskeletal structures, and its subcellular localization is dependent on microtubule morphology. Treatment of cells with nocodazole leads to dramatic reorganization of Bicd2, and correlates with Nek8 phosphorylation. This may be indicative of a role for Nek8 and Bicd2 associated with cell cycle independent microtubule dynamics. PMID- 11864969 TI - The role of ATP hydrolysis for kinesin processivity. AB - Conventional kinesin is a highly processive, plus-end-directed microtubule-based motor that drives membranous organelles toward the synapse in neurons. Although recent structural, biochemical, and mechanical measurements are beginning to converge into a common view of how kinesin converts the energy from ATP turnover into motion, it remains difficult to dissect experimentally the intermolecular domain cooperativity required for kinesin processivity. We report here our pre steady-state kinetic analysis of a kinesin switch I mutant at Arg(210) (NXXSSRSH, residues 205-212 in Drosophila kinesin). The results show that the R210A substitution results in a dimeric kinesin that is defective for ATP hydrolysis and a motor that cannot detach from the microtubule although ATP binding and microtubule association occur. We propose a mechanistic model in which ATP binding at head 1 leads to the plus-end-directed motion of the neck linker to position head 2 forward at the next microtubule binding site. However, ATP hydrolysis is required at head 1 to lock head 2 onto the microtubule in a tight binding state before head 1 dissociation from the microtubule. This mechanism optimizes forward movement and processivity by ensuring that one motor domain is tightly bound to the microtubule before the second can detach. PMID- 11864970 TI - Disruption of choline methyl group donation for phosphatidylethanolamine methylation in hepatocarcinoma cells. AB - Despite being widely hypothesized, the actual contribution of choline as a methyl source for phosphatidylethanolamine (PE) methylation has never been demonstrated, mainly due to the inability of conventional methods to distinguish the products from that of the CDP-choline pathway. Using a novel combination of stable-isotope labeling and tandem mass spectrometry, we demonstrated for the first time that choline contributed to phosphatidylcholine (PC) synthesis both as an intact choline moiety via the CDP-choline pathway and as a methyl donor via PE methylation pathway. When hepatocytes were labeled with d(9)-choline containing three deuterium atoms on each of the three methyl groups, d(3)-PC and d(6)-PC were detected, indicating that newly synthesized PC contained one or more individually mobilized methyl groups from d(9)-choline. The synthesis of d(3)-PC and d(6)-PC was sensitive to the general methylation inhibitor 3-deazaadenosine and were specific products of PE methylation using choline as a one-carbon donor. While the contribution to the CDP-choline pathway remained intact in hepatocarcinoma cells, contribution of choline to PE methylation was completely disrupted. In addition to a previously identified lack of PE methyltransferase, hepatocarcinoma cells were found to lack the abilities to oxidize choline to betaine and to donate the methyl group from betaine to homocysteine, whereas the usage of exogenous methionine as a methyl group donor was normal. The failure to use choline as a methyl source in hepatocarcinoma cells may contribute to methionine dependence, a widely observed aberration of one-carbon metabolism in malignancy. PMID- 11864972 TI - Salt-inducible kinase represses cAMP-dependent protein kinase-mediated activation of human cholesterol side chain cleavage cytochrome P450 promoter through the CREB basic leucine zipper domain. AB - Salt-inducible kinase (SIK), one of the serine/threonine protein kinases, was transiently expressed in Y1 cells during the early phase of the ACTH/cAMP dependent protein kinase (PKA)-mediated signal transduction. The overexpression of SIK(N), the SIK's N-terminal kinase domain, repressed the expression of the side chain cleavage cytochrome P450 (CYP11A) gene. To elucidate the mechanism of the repression by SIK, several CYP11A promoter constructs were tested for the promoter activities in the presence of PKA and/or SIK(N). A cAMP-response element (CRE)-like sequence present in the promoter was shown to be responsible not only for the PKA-mediated promoter activation but also for the SIK(N)-mediated repression. When the Gal4 DNA binding domain-linked full-length CRE-binding protein (CREB) construct was cotransfected with Gal4 reporter gene, SIK(N) repressed the PKA-induced reporter gene expression. However, SIK(N) could not repress the PKA-induced reporter activity conferred by Gal4 DNA binding domain linked basic leucine zipper (bZIP)-less CREB or bZIP-disrupted CREB. On the other hand, SIK(N) could repress the kinase-inducible domain-disrupted CREB-dependent reporter gene expression in the presence of PKA. The in vitro kinase reaction studies showed that SIK(N) could not phosphorylate CREB, and PKA failed to phosphorylate SIK(N). Taken together, these results suggest that SIK(N), cooperating with PKA, may act on the CREB's bZIP domain and repress the CREB mediated transcriptional activation of the CYP11A gene. PMID- 11864971 TI - Calcium-dependent involucrin expression is inversely regulated by protein kinase C (PKC)alpha and PKCdelta. AB - Calcium is an important physiologic regulator of keratinocyte function that may regulate keratinocyte differentiation via modulation of protein kinase C (PKC) activity. PKCalpha and PKCdelta are two PKC isoforms that are expressed at high levels in keratinocytes. In the present study, we examine the effect of PKCdelta and PKCalpha on calcium-dependent keratinocyte differentiation as measured by effects on involucrin (hINV) gene expression. Our studies indicate that calcium increases hINV promoter activity and endogenous hINV gene expression. This response requires PKCdelta, as evidenced by the observation that treatment with dominant-negative PKCdelta inhibits calcium-dependent hINV promoter activity, whereas wild type PKCdelta increases activity. PKCalpha, in contrast, inhibits calcium-dependent hINV promoter activation, a finding that is consistent with the ability of dominant-negative PKCalpha and the PKCalpha inhibitor, Go6976, to increase hINV gene expression. The calcium-dependent regulatory response is mediated by an AP1 transcription factor-binding site located within the hINV promoter distal regulatory region that is also required for PKCdelta-dependent regulation; moreover, both calcium and PKCdelta produce similar, but not identical, changes in AP1 factor expression. A key question is whether calcium directly influences PKC isoform function. Our studies show that calcium does not regulate PKCalpha or delta levels or cause a marked redistribution to membranes. However, tyrosine phosphorylation of PKCdelta is markedly increased following calcium treatment. These findings suggest that PKCalpha and PKCdelta are required for, and modulate, calcium-dependent keratinocyte differentiation in opposing directions. PMID- 11864973 TI - Ubiquitin/proteasome-dependent degradation of D-type cyclins is linked to tumor necrosis factor-induced cell cycle arrest. AB - Tumor necrosis factor-alpha (TNF) is well known for its cytotoxic effect on malignant cells. Its role in cell cycle control is relatively less known. In this study, we found that TNF induced G(1) arrest of TF-1 and MV4-11 cells while simultaneously causing apoptosis. Treatment of the cells with TNF for 48 h caused cell cycle arrest, accompanied by dephosphorylation of pRb and reduction in D type cyclin expression. The down-regulation of the D-type cyclins resulted in approximately 50-80% decrease of the cyclin-dependent kinase activities. Cells treated with calpain-dependent inhibitor ALLN and apoptosis inhibitor zVAD-FMK suppressed degradation of IkappaBalpha and activation of caspase 3, respectively. However, treatment of cells with these two inhibitors was not able to prevent TNF induced down-regulation of the D-type cyclins. In contrast, proteasome inhibitor MG-132 and lactacystin blocked both TNF-induced degradation of IkappaBalpha and down-regulation of D-type cyclins. These data suggest that down-regulation of D type cyclins by TNF may be proteasome-proteolysis dependent. Additional support for this conclusion was obtained from experiments showing an increase of proteasome activity in TNF-treated cells and in vitro degradation of cyclin D3 by 26 S proteasome. PMID- 11864974 TI - Minor alteration of microtubule dynamics causes loss of tension across kinetochore pairs and activates the spindle checkpoint. AB - We have previously identified the opium alkaloid noscapine as a microtubule interacting agent that binds stoichiometrically to tubulin and alters its conformation. Here we show that, unlike many other microtubule inhibitors, noscapine does not significantly promote or inhibit microtubule polymerization. Instead, it alters the steady-state dynamics of microtubule assembly, primarily by increasing the amount of time that the microtubules spend in an attenuated (pause) state. Further studies reveal that even at high concentrations, noscapine does not alter the tubulin polymer/monomer ratio in HeLa cells. Cells treated with noscapine arrest at mitosis with nearly normal bipolar spindles. Strikingly, although most of the chromosomes in these cells are aligned at the metaphase plate, the rest remain near the spindle poles, both of which exhibit loss of tension across kinetochore pairs. Furthermore, levels of the spindle checkpoint proteins Mad2, Bub1, and BubR1 decrease by 138-, 3.7-, and 3.9-fold, respectively, at the kinetochore region upon chromosome alignment. Our results thus suggest that an exquisite control of microtubule dynamics is required for kinetochore tension generation and chromosome alignment during mitosis. Our data also support the idea that Mad2 and Bub1/BubR1 respond to kinetochore-microtubule attachment and/or tension to different degrees. PMID- 11864975 TI - Differential characteristics and subcellular localization of two starch-branching enzyme isoforms encoded by a single gene in Phaseolus vulgaris L. AB - Starch-branching enzymes (SBE) have a dominant role for amylopectin structure as they define chain length and frequency of branch points. We have previously shown that one of the SBE isoforms of kidney bean (Phaseolus vulgaris L.), designated PvSBE2, has a molecular mass (82 kDa) significantly smaller than those reported for isologous SBEs from pea (SBEI), maize (BEIIb), and rice (RBE3). Additionally, in contrast to the dual location of the pea SBEI in both the soluble and starch granule fractions, PvSBE2 was found only in the soluble fraction during seed development. Analysis of a pvsbe2 cDNA suggested that PvSBE2 is generated from a larger precursor with a putative plastid targeting sequence of 156 residues. Here we describe the occurrence of a larger 100-kDa form (LF-PvSBE2) of PvSBE2 found both in the soluble and starch granule fractions of the developing seeds. The determined N-terminal sequence, VKSSHDSD, of LF-PvSBE2 corresponded to a peptide sequence located 111 amino acids upstream from the N terminus of purified PvSBE2, suggesting that LF-PvSBE2 and PvSBE2 are products of the same gene. Analysis of the products by 5'-RACE (rapid amplification of cDNA ends) and reverse transcription PCR indicated that the two transcripts for pre-LF-PvSBE2 and pre PvSBE2 are generated by alternative splicing. Recombinant LF-PvSBE2 (rLF-PvSBE2) was purified from Escherichia coli and the kinetic properties were compared with those of recombinant PvSBE2 (rPvSBE2). rLF-PvSBE2 had much higher affinity for amylopectin (K(m) = 4.4 mg/ml) than rPvSBE2 (18.4 mg/ml), whereas the V(max) of rLF-PvSBE2 (135 units/mg) for this substrate was much lower than that of rPvSBE2 (561 units/mg). These results suggest that the N-terminal extension of LF-PvSBE2 plays a critical role for localization in starch granules by altering its enzymatic properties. PMID- 11864976 TI - The course of etoposide-induced apoptosis from damage to DNA and p53 activation to mitochondrial release of cytochrome c. AB - Treatment of L929 fibroblasts by the topoisomerase II inhibitor etoposide killed 50% of the cells within 72 h. The cell killing was preceded by the release of cytochrome c from the mitochondria. Simultaneous treatment of the cells with wortmannin, cycloheximide, furosemide, cyclosporin A, or decylubiquinone prevented the release of cytochrome c and significantly reduced the loss of viability. Etoposide caused the phosphorylation of p53 within 6 h, an effect prevented by wortmannin, an inhibitor of DNA-dependent protein kinase (DNA-PK). The activation of p53 by etoposide resulted in the up-regulation of the pro apoptotic protein Bax, a result that was prevented by the protein synthesis inhibitor cycloheximide. The increase in the content of Bax was followed by the translocation of this protein from the cytosol to the mitochondria, an event that was inhibited by furosemide, a chloride channel inhibitor. Stably transfected L929 fibroblasts that overexpress Akt were resistant to etoposide and did not translocate Bax to the mitochondria or release cytochrome c. Bax levels in these transfected cells were comparable with the wild-type cells. The release of cytochrome c upon translocation of Bax has been attributed to induction of the mitochondrial permeability transition (MPT). Cyclosporin A and decylubiquinone, inhibitors of MPT, prevented the release of cytochrome c without affecting Bax translocation. These data define a sequence of biochemical events that mediates the apoptosis induced by etoposide. This cascade proceeds by coupling DNA damage to p53 phosphorylation through the action of DNA-PK. The activation of p53 increases Bax synthesis. The translocation of Bax to the mitochondria induces the MPT, the event that releases cytochrome c and culminates in the death of the cells. PMID- 11864977 TI - Pheromone-dependent ubiquitination of the mitogen-activated protein kinase kinase Ste7. AB - Many cell signaling pathways are regulated by phosphorylation, ubiquitination, and degradation of constituent proteins. As with phosphorylation, protein ubiquitination can be reversed, through the action of ubiquitin-specific processing proteases (UBPs). Here we have analyzed 15 UBP disruption mutants in the yeast Saccharomyces cerevisiae and identified one (ubp3 Delta) that acts specifically in the pheromone response pathway. Upon pheromone stimulation, ubp3 Delta mutants accumulate unconjugated polyubiquitin chains as well as polyubiquitinated forms of the mitogen-activated protein kinase kinase Ste7. The ubp3 Delta mutants exhibit a potentiated response to pheromone, as measured by in vivo MAP kinase activity, transcriptional induction, and cell cycle arrest. Signaling is likewise enhanced upon direct activation of Ste4 (G protein beta subunit) and Ste11 (Ste7 kinase) but not the downstream transcription factor Ste12. These findings reveal a mechanism by which pheromone-triggered ubiquitination of Ste7 can modulate the pheromone response in vivo. PMID- 11864978 TI - P2X4 receptor is a glycosylated cardiac receptor mediating a positive inotropic response to ATP. AB - Although P2X receptors are suggested to play a role in synaptic neurotransmission, the specific physiological role of each P2X receptor subtype remains largely unknown. We used cultured chick embryo ventricular myocytes as a model to study a potential physiological role of the P2X(4) receptor in mediating the positive inotropic effect of ATP. The chick P2X4 receptor (cP2X(4)R) mRNA was expressed in the heart and the pharmacological features of the ATP-induced positive inotropic response were similar to those of the cP2X(4)R in terms of insensitivity to blockade by known P2 receptor antagonists and the ineffectiveness of adenosine 5'-(alpha,beta-methylene)triphosphate as an agonist. Treatment of myocytes with antisense oligonucleotides specific to the 5' region of cP2X(4)R abrogated the P2 agonist-stimulated (45)Ca influx. Similarly, antisense oligonucleotide treatment also blocked the 2-methylthio-ATP-stimulated increase in contractile amplitude. The data suggest that the native P2X(4) receptor is involved in mediating the P2 agonist-stimulated response in the heart. In characterizing the biochemical property of the P2X(4) receptor, antibody against cP2X(4)R detected a 44-kDa and a 58-kDa protein in the immunoblot. Inhibition of N-linked glycosylation by tunicamycin converted the 58 kDa protein to the 44-kDa protein, suggesting that the 58-kDa protein was a glycosylated P2X(4) receptor. The nonglycosylated 44-kDa P2X(4) receptor was resistant to various detergent/aqueous extraction, consistent with a role of glycosylation in maintaining its detergent solubility and hydrophilicity. Cross linking the cell surface proteins with N-hydroxysuccinimide-SS-biotin followed by affinity precipitation with streptavidin-conjugated agarose and subsequent immunoblotting with anti-cP2X(4)R showed that only the glycosylated 58-kDa P2X(4) receptor was expressed on the cell surface, indicating an important role of glycosylation for the receptor's localization on the plasma membrane. These data revealed a novel physiologic function of the P2X(4) receptor and suggested the importance of N-linked glycosylation in its cell surface expression and detergent solubility. PMID- 11864979 TI - Effect of hyaluronan oligosaccharides on the expression of heat shock protein 72. AB - We have previously shown that intraarticular treatment with a hyaluronan (HA) preparation (840 kDa), HA84, up-regulates heat shock protein 72 (Hsp72) expression and suppresses degeneration of synovial cells in an arthritis model. In that study, the HA84 administered was degraded into HA oligosaccharides in the synovial tissue, suggesting that HA84 or degradation products of HA may up regulate Hsp72 expression. Thus, in the present study, we examined the effects of HA of various molecular sizes on Hsp72 expression and cell death in stressed cells. Western blotting analysis showed that treatment of K562 cells with HA tetrasaccharides up-regulated Hsp72 expression after exposure to hyperthermia. On the other hand, treatment of the cells with HA of other sizes (di-, hexa-, deca-, dodecasaccharides), HA84, or tetrasaccharides of keratan sulfate did not elicit any change in expression of the Hsp72 protein. Treatment of the cells with tetrasaccharides of HA up-regulated not only expression of the Hsp72 protein but also Hsp72 mRNA expression and enhanced activation of HSF1, a transcription factor controlling Hsp72 expression, after exposure to hyperthermia. Because the level of Hsp72 protein was not affected by tetrasaccharides of HA when the K562 cells were kept at 37 degrees C without any stress, it is evident that tetrasaccharides of HA did not act as a stress factor. In addition, tetrasaccharides of HA suppressed cell death in the case of K562 cells exposed to hyperthermia and of PC12 cells under serum deprivation. These results suggest that a certain size of oligosaccharides, i.e. the tetrasaccharides of HA, up regulates Hsp72 expression by enhancing the activation of HSF1 under stress conditions and suppresses cell death. PMID- 11864980 TI - The high specificities of Phaseolus vulgaris erythro- and leukoagglutinating lectins for bisecting GlcNAc or beta 1-6-linked branch structures, respectively, are attributable to loop B. AB - Despite very similar tertiary structures based upon a common framework, legume lectins exhibit an amazing variety of sugar binding specificities. While most of these lectins recognize rather discrete sugar linkages, Phaseolus vulgaris erythroagglutinating and leukoagglutinating lectins (E(4)- and L(4)-PHA) are unique in recognizing larger structures. E(4)- and L(4)-PHA are known to recognize complex type N-glycans containing bisecting GlcNAc or a beta1,6-linked branch, respectively. However, the detailed mechanisms of molecular recognition are poorly understood. In order to dissect the contributions of different portions of each lectin, we carried out region-swapping mutagenesis between E(4)- and L(4)-PHA. We prepared six chimeric lectins by exchanging different combinations of loop B and the central portion of loop C, two of four loops thought to be important for the recognition of monosaccharides (Sharma, V., and Surolia, A. (1997) J. Mol. Biol. 267, 433-445). The chimeric lectins' sugar binding activities were evaluated quantitatively by surface plasmon resonance. These comparisons indicate that the high specificities of E(4)- and L(4)-PHA toward bisecting GlcNAc and beta1,6-linked branch structures are almost solely attributable to loop B. The contribution of the central portion of loop C to the recognition of those structural motifs was found to be negligible. Instead, it modulates affinity toward LacNAc residues present at the nonreducing terminus. Moreover, some of the chimeric lectins prepared in this study showed even higher specificities/affinities than native E(4)- and L(4)-PHA toward complex sugar chains containing either a bisecting GlcNAc residue or a beta1,6-linked branch. PMID- 11864981 TI - Distinct functions of the unique C terminus of LAP2alpha in cell proliferation and nuclear assembly. AB - The non-membrane-bound lamina-associated polypeptide 2 isoform, LAP2alpha, forms nucleoskeletal structures with A-type lamins and interacts with chromosomes in a cell cycle-dependent manner. LAP2alpha contains a LEM (LAP2, emerin, and MAN1) domain in the constant N terminus that binds to chromosomal barrier-to autointegration factor, and a C-terminal unique region that is essential for chromosome binding. Here we show that C-terminal LAP2alpha fragment efficiently bound to mitotic chromosomes and inhibited assembly of endogenous LAP2alpha, nuclear membranes, and lamins A/C in in vitro nuclear assembly assays. Full length recombinant LAP2alpha, which bound to chromosomes, and N-terminal fragment, which did not bind, had no effect on assembly. This suggested an essential role for the LAP2alpha C terminus in chromosome association and for the N-terminal LEM domain in subsequent assembly stages. In vivo analysis upon transient expression of GFP-tagged LAP2alpha fragments confirmed that, unlike the N-terminal fragment, the C-terminal fragment was able to bind to chromosomes during mitosis, if expressed weakly. At higher expression levels, C-terminal LAP2alpha fragment and full-length protein led to cell cycle arrest in interphase and apoptosis, as shown by fluorescence-activated cell sorter analysis, time lapse microscopy, and BrdUrd incorporation assays. These data indicated distinct functions of LAP2alpha in cell cycle progression during interphase and in nuclear reassembly during mitosis. PMID- 11864982 TI - Oxygen adaptation. The role of the CcoQ subunit of the cbb3 cytochrome c oxidase of Rhodobacter sphaeroides 2.4.1. AB - The cbb(3) cytochrome c oxidase of Rhodobacter sphaeroides consists of four nonidentical subunits. Three subunits (CcoN, CcoO, and CcoP) comprise the catalytic "core" complex required for the reduction of O(2) and the oxidation of a c-type cytochrome. On the other hand, the functional role of subunit IV (CcoQ) of the cbb(3) oxidase was not obvious, although we previously suggested that it is involved in the signal transduction pathway controlling photosynthesis gene expression (Oh, J. I., and Kaplan, S. (1999) Biochemistry 38, 2688-2696). Here we go on to demonstrate that subunit IV protects the core complex, in the presence of O(2), from proteolytic degradation by a serine metalloprotease. In the absence of CcoQ, we suggest that the presence of O(2) leads to the loss of heme from the core complex, which destabilizes the cbb(3) oxidase into a "degradable" form, perhaps by altering its conformation. Under aerobic conditions the absence of CcoQ appears to affect the CcoP subunit most severely. It was further demonstrated, using a series of COOH-terminal deletion derivatives of CcoQ, that the minimum length of CcoQ required for stabilization of the core complex under aerobic conditions is the amino-terminal approximately 48-50 amino acids. PMID- 11864984 TI - Interactions between S4-S5 linker and S6 transmembrane domain modulate gating of HERG K+ channels. AB - Outward movement of the voltage sensor is coupled to activation in voltage-gated ion channels; however, the precise mechanism and structural basis of this gating event are poorly understood. Potential insight into the coupling mechanism was provided by our previous finding that mutation to Lys of a single residue (Asp(540)) located in the S4-S5 linker endowed HERG (human ether-a-go-go-related gene) K(+) channels with the unusual ability to open in response to membrane depolarization and hyperpolarization in a voltage-dependent manner. We hypothesized that the unusual hyperpolarization-induced gating occurred through an interaction between Lys(540) and the C-terminal end of the S6 domain, the region proposed to form the activation gate. Therefore, we mutated six residues located in this region of S6 (Ile(662)-Tyr(667)) to Ala in D540K HERG channels. Mutation of Arg(665), but not the other five residues, prevented hyperpolarization-dependent reopening of D540K HERG channels. Mutation of Arg(665) to Gln or Asp also prevented reopening. In addition, D540R and D540K/R665K HERG reopened in response to hyperpolarization. Together these findings suggest that a single residue (Arg(665)) in the S6 domain interacts with Lys(540) by electrostatic repulsion to couple voltage sensing to hyperpolarization-dependent opening of D540K HERG K(+) channels. Moreover, our findings suggest that the C-terminal ends of S4 and S6 are in close proximity at hyperpolarized membrane potentials. PMID- 11864983 TI - Desaturation and hydroxylation. Residues 148 and 324 of Arabidopsis FAD2, in addition to substrate chain length, exert a major influence in partitioning of catalytic specificity. AB - Exchanging the identity of amino acids at four key locations within the Arabidopsis thaliana oleate desaturase (FAD2) and the Lesquerella fendleri hydroxylase/desaturase (LFAH) was shown to influence partitioning between desaturation and hydroxylation (Broun, P., Shanklin, J., Whittle, E., and Somerville, C. (1998) Science 282, 1315-1317). We report that four analogous substitutions in the FAD2 sequence by their equivalents from the castor oleate hydroxylase result in hydroxy fatty acid accumulation in A. thaliana to the same levels as for the wild-type castor hydroxylase. We also describe the relative contribution of these substitutions, both individually and in combination, by analyzing the products resulting from their expression in A. thaliana and/or Saccharomyces cerevisiae. Yeast expression showed that M324V, a change reachable by a single point mutation, altered the product distribution approximately 49 fold, and that residue 148 is also a predominant determinant of reaction outcome. Comparison of residues at position 148 of FAD2, LFAH, and the Ricinus oleate hydroxylase prompted us to rationally engineer LFAH-N149I, a variant with approximately 1.9-fold increase in hydroxylation specificity compared with that of wild-type LFAH. Control experiments showed that the wild-type Arabidopsis thaliana FAD2 desaturase has inherent, low level, hydroxylation activity. Further, fatty acid desaturases from different kingdoms and with different regiospecificities exhibit similar intrinsic hydroxylase activity, underscoring fundamental mechanistic similarities between desaturation and hydroxylation. For LFAH mutants the hydroxylation:desaturation ratio is 5-9-fold higher for 18 carbon versus 16-carbon substrates, supporting our hypothesis that substrate positioning in the active site plays a key role in the partitioning of catalytic specificity. PMID- 11864985 TI - Mapping the binding site of a human ether-a-go-go-related gene-specific peptide toxin (ErgTx) to the channel's outer vestibule. AB - The goals of this study are to investigate the mechanism and site of action whereby a human ether-a-go-go-related gene (HERG)-specific scorpion peptide toxin, ErgTx, suppresses HERG current. We apply cysteine-scanning mutagenesis to the S5-P and P-S6 linkers of HERG and examine the resulting changes in ErgTx potency. Data are compared with the characteristics of charybdotoxin (ChTx, or its analogs) binding to the Shaker channel. ErgTx binds to the outer vestibule of HERG but may not physically occlude the pore. In contrast to ChTx. Shaker interaction, elevating [K](o) (from 2 to 98 mm) does not affect ErgTx potency, and through-solution electrostatic forces only play a minor role in influencing ErgTx.HERG interaction. Cysteine mutations of three positions in S5-P linker (Trp 585, Gly-590, and Ile-593) and 1 position in P-S6 linker (Pro-632) induce profound changes in ErgTx binding (DeltaDeltaG > 2 kcal/mol). We propose that the long S5-P linker of the HERG channel forms an amphipathic alpha-helix that, together with the P-S6 linker, forms a hydrophobic ErgTx binding site. This study paves the way for future mutant cycle analysis of interacting residues in the ErgTx.HERG complex, which, in conjunction with NMR determination of the ErgTx solution structure, will yield information about the topology of HERG's outer vestibule. PMID- 11864986 TI - Prometastatic effect of N-acetylglucosaminyltransferase V is due to modification and stabilization of active matriptase by adding beta 1-6 GlcNAc branching. AB - Oligosaccharide moieties of glycoproteins are structurally altered during development, carcinogenesis, and malignant transformations. It is well known that beta1-6 GlcNAc branching, a product of UDP-GlcNAc alpha-mannoside beta1-6-N acetylglucosaminyltransferase (GnT-V), is associated with malignant transformation as the results of such alterations. However, the mechanism by which beta1-6 GlcNAc branching is linked to metastasis remains unclear, because the identification of specific glycoprotein(s) that are glycosylated by GnT-V and its biological function have not been examined. We herein report that matriptase, which activates both urokinase-type plasminogen activator and hepatocyte growth factor, is a target protein for GnT-V. The overexpression of GnT-V in gastric cancer cells leads to severe peritoneal dissemination in athymic mice, which can be attributed to the increased expression of matriptase. This increase was due to the acquired resistance of matriptase to degradation, since it is glycosylated by GnT-V and a corresponding increase in the active form. These results indicate that this process is a key element in malignant transformation, as the direct result of oligosaccharide modification. PMID- 11864987 TI - Oxyresveratrol and hydroxystilbene compounds. Inhibitory effect on tyrosinase and mechanism of action. AB - Tyrosinase is responsible for the molting process in insects, undesirable browning of fruits and vegetables, and coloring of skin, hair, and eyes in animals. To clarify the mechanism of the depigmenting property of hydroxystilbene compounds, inhibitory actions of oxyresveratrol and its analogs on tyrosinases from mushroom and murine melanoma B-16 have been elucidated in this study. Oxyresveratrol showed potent inhibitory effect with an IC(50) value of 1.2 microm on mushroom tyrosinase activity, which was 32-fold stronger inhibition than kojic acid, a depigmenting agent used as the cosmetic material with skin-whitening effect and the medical agent for hyperpigmentation disorders. Hydroxystilbene compounds of resveratrol, 3,5-dihydroxy-4'-methoxystilbene, and rhapontigenin also showed more than 50% inhibition at 100 microm on mushroom tyrosinase activity, but other methylated or glycosylated hydroxystilbenes of 3,4'-dimethoxy 5-hydroxystilbene, trimethylresveratrol, piceid, and rhaponticin did not inhibit significantly. None of the hydroxystilbene compounds except oxyresveratrol exhibited more than 50% inhibition at 100 microm on l-tyrosine oxidation by murine tyrosinase activity; oxyresveratrol showed an IC(50) value of 52.7 microm on the enzyme activity. The kinetics and mechanism for inhibition of mushroom tyrosinase exhibited the reversibility of oxyresveratrol as a noncompetitive inhibitor with l-tyrosine as the substrate. The interaction between oxyresveratrol and tyrosinase exhibited a high affinity reflected in a K(i) value of 3.2-4.2 x 10(-7) m. Oxyresveratrol did not affect the promoter activity of the tyrosinase gene in murine melanoma B-16 at 10 and 100 microm. Therefore, the depigmenting effect of oxyresveratrol works through reversible inhibition of tyrosinase activity rather than suppression of the expression and synthesis of the enzyme. The number and position of hydroxy substituents seem to play an important role in the inhibitory effects of hydroxystilbene compounds on tyrosinase activity. PMID- 11864988 TI - Binding to Elongin C inhibits degradation of interacting proteins in yeast. AB - Elongin C is a highly conserved, low molecular weight protein found in a variety of multiprotein complexes in human, rat, fly, worm, and yeast cells. Among the best characterized of these complexes is a mammalian E3 ligase that targets proteins for ubiquitination and subsequent degradation by the 26 S proteasome. Despite its crucial role as a component of such E3 ligases and other complexes, the specific function of Elongin C is unknown. In yeast, Elongin C is a non essential gene and there is no obvious phenotype as associated with its absence. We previously reported that in Saccharomyces cerevisiae Elongin C (Elc1) interacts specifically and strongly with a class of proteins loosely defined as stress response proteins. In the present study, we examined the role of yeast Elc1 in the turnover of two of these binding partners, Snf4 and Pcl6. Deletion of Elc1 resulted in decreased steady-state levels of Snf4 and Pcl6 as indicated by Western blot analysis. Northern blot analysis of mRNA prepared from elc1 null and wild type strains revealed no difference in mRNA levels for Snf4 and Pcl6 establishing that the effects of Elc1 are not transcriptionally mediated. Reintroduction of either yeast or human Elongin C into Elc1 null strains abrogated this effect. Taken together, these data document that the levels of Snf4 and Pcl6 are dependent on the presence of Elc1 and that binding to Elc1 inhibits the degradation of these proteins. The results suggest a new function for yeast Elongin C that is distinct from a direct role in targeting proteins for ubiquitination and subsequent proteolysis. PMID- 11864989 TI - Polyunsaturated fatty acyl coenzyme A suppress the glucose-6-phosphatase promoter activity by modulating the DNA binding of hepatocyte nuclear factor 4 alpha. AB - Glucose-6-phosphatase confers on gluconeogenic tissues the capacity to release endogenous glucose in blood. The expression of its gene is modulated by nutritional mechanisms dependent on dietary fatty acids, with specific inhibitory effects of polyunsaturated fatty acids (PUFA). The presence of consensus binding sites of hepatocyte nuclear factor 4 (HNF4) in the -1640/+60 bp region of the rat glucose-6-phosphatase gene has led us to consider the hypothesis that HNF4 alpha could be involved in the regulation of glucose-6-phosphatase gene transcription by long chain fatty acid (LCFA). Our results have shown that the glucose-6 phosphatase promoter activity is specifically inhibited in the presence of PUFA in HepG2 hepatoma cells, whereas saturated LCFA have no effect. In HeLa cells, the glucose-6-phosphatase promoter activity is induced by the co-expression of HNF4 alpha or HNF1 alpha. PUFA repress the promoter activity only in HNF4 alpha cotransfected HeLa cells, whereas they have no effects on the promoter activity in HNF1 alpha-cotransfected HeLa cells. From gel shift mobility assays, deletion, and mutagenesis experiments, two specific binding sequences have been identified that appear able to account for both transactivation by HNF4 alpha and regulation by LCFA in cells. The binding of HNF4 alpha to its cognate sites is specifically inhibited by polyunsaturated fatty acyl coenzyme A in vitro. These data strongly suggest that the mechanism by which PUFA suppress the glucose-6-phosphatase gene transcription involves an inhibition of the binding of HNF4 alpha to its cognate sites in the presence of polyunsaturated fatty acyl-CoA thioesters. PMID- 11864990 TI - Quantitative determination of binding affinity of delta-subunit in Escherichia coli F1-ATPase: effects of mutation, Mg2+, and pH on Kd. AB - To study the stator function in ATP synthase, a fluorimetric assay has been devised for quantitative determination of binding affinity of delta-subunit to Escherichia coli F(1)-ATPase. The signal used is that of the natural tryptophan at residue delta28, which is enhanced by 50% upon binding of delta-subunit to alpha(3)beta(3)gammaepsilon complex. K(d) for delta binding is 1.4 nm, which is energetically equivalent (50.2 kJ/mol) to that required to resist the rotor strain. Only one site for delta binding was detected. The deltaW28L mutation increased K(d) to 4.6 nm, equivalent to a loss of 2.9 kJ/mol binding energy. While this was insufficient to cause detectable functional impairment, it did facilitate preparation of delta-depleted F(1). The alphaG29D mutation reduced K(d) to 26 nm, equivalent to a loss of 7.2 kJ/mol binding energy. This mutation did cause serious functional impairment, referable to interruption of binding of delta to F(1). Results with the two mutants illuminate how finely balanced is the stator resistance function. delta' fragment, consisting of residues delta1-134, bound with the same K(d) as intact delta, showing that, at least in absence of F(o) subunits, the C-terminal domain of delta contributes zero binding energy. Mg(2+) ions had a strong effect on increasing delta binding affinity, supporting the possibility of bridging metal ion involvement in stator function. High pH environment greatly reduced delta binding affinity, suggesting the involvement of protonatable side-chains in the binding site. PMID- 11864991 TI - Interactions and regulation of molecular motors in Xenopus melanophores. AB - Many cellular components are transported using a combination of the actin- and microtubule-based transport systems. However, how these two systems work together to allow well-regulated transport is not clearly understood. We investigate this question in the Xenopus melanophore model system, where three motors, kinesin II, cytoplasmic dynein, and myosin V, drive aggregation or dispersion of pigment organelles called melanosomes. During dispersion, myosin V functions as a "molecular ratchet" to increase outward transport by selectively terminating dynein-driven minus end runs. We show that there is a continual tug-of-war between the actin and microtubule transport systems, but the microtubule motors kinesin II and dynein are likely coordinated. Finally, we find that the transition from dispersion to aggregation increases dynein-mediated motion, decreases myosin V--mediated motion, and does not change kinesin II--dependent motion. Down-regulation of myosin V contributes to aggregation by impairing its ability to effectively compete with movement along microtubules. PMID- 11864992 TI - Ubiquitination and proteasomal activity is required for transport of the EGF receptor to inner membranes of multivesicular bodies. AB - EGF, but not TGF alpha, efficiently induces degradation of the EGF receptor (EGFR). We show that EGFR was initially polyubiquitinated to the same extent upon incubation with EGF and TGF alpha, whereas the ubiquitination was more sustained by incubation with EGF than with TGF alpha. Consistently, the ubiquitin ligase c Cbl was recruited to the plasma membrane upon activation of the EGFR with EGF and TGF alpha, but localized to endosomes only upon activation with EGF. EGF remains bound to the EGFR upon endocytosis, whereas TGF alpha dissociates from the EGFR. Therefore, the sustained polyubiquitination is explained by EGF securing the kinase activity of endocytosed EGFR. Overexpression of the dominant negative N Cbl inhibited ubiquitination of the EGFR and degradation of EGF and EGFR. This demonstrates that EGF-induced ubiquitination of the EGFR as such is important for lysosomal sorting. Both lysosomal and proteasomal inhibitors blocked degradation of EGF and EGFR, and proteasomal inhibitors inhibited translocation of activated EGFR from the outer limiting membrane to inner membranes of multivesicular bodies (MVBs). Therefore, lysosomal sorting of kinase active EGFR is regulated by proteasomal activity. Immuno-EM showed the localization of intact EGFR on internal membranes of MVBs. This demonstrates that the EGFR as such is not the proteasomal target. PMID- 11864993 TI - PKC alpha regulates the hypertrophic growth of cardiomyocytes through extracellular signal-regulated kinase1/2 (ERK1/2). AB - Members of the protein kinase C (PKC) isozyme family are important signal transducers in virtually every mammalian cell type. Within the heart, PKC isozymes are thought to participate in a signaling network that programs developmental and pathological cardiomyocyte hypertrophic growth. To investigate the function of PKC signaling in regulating cardiomyocyte growth, adenoviral mediated gene transfer of wild-type and dominant negative mutants of PKC alpha, beta II, delta, and epsilon (only wild-type zeta) was performed in cultured neonatal rat cardiomyocytes. Overexpression of wild-type PKC alpha, beta II, delta, and epsilon revealed distinct subcellular localizations upon activation suggesting unique functions of each isozyme in cardiomyocytes. Indeed, overexpression of wild-type PKC alpha, but not betaI I, delta, epsilon, or zeta induced hypertrophic growth of cardiomyocytes characterized by increased cell surface area, increased [(3)H]-leucine incorporation, and increased expression of the hypertrophic marker gene atrial natriuretic factor. In contrast, expression of dominant negative PKC alpha, beta II, delta, and epsilon revealed a necessary role for PKC alpha as a mediator of agonist-induced cardiomyocyte hypertrophy, whereas dominant negative PKC epsilon reduced cellular viability. A mechanism whereby PKC alpha might regulate hypertrophy was suggested by the observations that wild-type PKC alpha induced extracellular signal-regulated kinase1/2 (ERK1/2), that dominant negative PKC alpha inhibited PMA-induced ERK1/2 activation, and that dominant negative MEK1 (up-stream of ERK1/2) inhibited wild type PKC alpha-induced hypertrophic growth. These results implicate PKC alpha as a necessary mediator of cardiomyocyte hypertrophic growth, in part, through a ERK1/2-dependent signaling pathway. PMID- 11864994 TI - In vivo dissection of the chromosome condensation machinery: reversibility of condensation distinguishes contributions of condensin and cohesin. AB - The machinery mediating chromosome condensation is poorly understood. To begin to dissect the in vivo function(s) of individual components, we monitored mitotic chromosome structure in mutants of condensin, cohesin, histone H3, and topoisomerase II (topo II). In budding yeast, both condensation establishment and maintenance require all of the condensin subunits, but not topo II activity or phospho-histone H3. Structural maintenance of chromosome (SMC) protein 2, as well as each of the three non-SMC proteins (Ycg1p, Ycs4p, and Brn1p), was required for chromatin binding of the condensin complex in vivo. Using reversible condensin alleles, we show that chromosome condensation does not involve an irreversible modification of condensin or chromosomes. Finally, we provide the first evidence of a mechanistic link between condensin and cohesin function. A model discussing the functional interplay between cohesin and condensin is presented. PMID- 11864995 TI - Modulation of the F-actin cytoskeleton by c-Abl tyrosine kinase in cell spreading and neurite extension. AB - The nonreceptor tyrosine kinase encoded by the c-Abl gene has the unique feature of an F-actin binding domain (FABD). Purified c-Abl tyrosine kinase is inhibited by F-actin, and this inhibition can be relieved through mutation of its FABD. The c-Abl kinase is activated by physiological signals that also regulate the actin cytoskeleton. We show here that c-Abl stimulated the formation of actin microspikes in fibroblasts spreading on fibronectin. This function of c-Abl is dependent on kinase activity and is not shared by c-Src tyrosine kinase. The Abl dependent F-actin microspikes occurred under conditions where the Rho-family GTPases were inhibited. The FABD-mutated c-Abl, which is active in detached fibroblasts, stimulated F-actin microspikes independent of cell attachment. Moreover, FABD-mutated c-Abl stimulated the formation of F-actin branches in neurites of rat embryonic cortical neurons. The reciprocal regulation between F actin and the c-Abl tyrosine kinase may provide a self-limiting mechanism in the control of actin cytoskeleton dynamics. PMID- 11864996 TI - 14-3-3 transits to the nucleus and participates in dynamic nucleocytoplasmic transport. AB - 14-3-3 proteins regulate the cell cycle and prevent apoptosis by controlling the nuclear and cytoplasmic distribution of signaling molecules with which they interact. Although the majority of 14-3-3 molecules are present in the cytoplasm, we show here that in the absence of bound ligands 14-3-3 homes to the nucleus. We demonstrate that phosphorylation of one important 14-3-3 binding molecule, the transcription factor FKHRL1, at the 14-3-3 binding site occurs within the nucleus immediately before FKHRL1 relocalization to the cytoplasm. We show that the leucine-rich region within the COOH-terminal alpha-helix of 14-3-3, which had been proposed to function as a nuclear export signal (NES), instead functions globally in ligand binding and does not directly mediate nuclear transport. Efficient nuclear export of FKHRL1 requires both intrinsic NES sequences within FKHRL1 and phosphorylation/14-3-3 binding. Finally, we present evidence that phosphorylation/14-3-3 binding may also prevent FKHRL1 nuclear reimport. These results indicate that 14-3-3 can mediate the relocalization of nuclear ligands by several mechanisms that ensure complete sequestration of the bound 14-3-3 complex in the cytoplasm. PMID- 11864997 TI - Trypanin is a cytoskeletal linker protein and is required for cell motility in African trypanosomes. AB - The cytoskeleton of eukaryotic cells is comprised of a complex network of distinct but interconnected filament systems that function in cell division, cell motility, and subcellular trafficking of proteins and organelles. A gap in our understanding of this dynamic network is the identification of proteins that connect subsets of cytoskeletal structures. We previously discovered a family of cytoskeleton-associated proteins that includes GAS11, a candidate human tumor suppressor upregulated in growth-arrested cells, and trypanin, a component of the flagellar cytoskeleton of African trypanosomes. Although these proteins are intimately associated with the cytoskeleton, their function has yet to be determined. Here we use double-stranded RNA interference to block trypanin expression in Trypanosoma brucei, and demonstrate that this protein is required for directional cell motility. Trypanin(minus sign) mutants have an active flagellum, but are unable to coordinate flagellar beat. As a consequence, they spin and tumble uncontrollably, occasionally moving backward. Immunofluorescence experiments demonstrate that trypanin is located along the flagellum/flagellum attachment zone and electron microscopic analysis revealed that cytoskeletal connections between the flagellar apparatus and subpellicular cytoskeleton are destabilized in trypanin(minus sign) mutants. These results indicate that trypanin functions as a cytoskeletal linker protein and offer insights into the mechanisms of flagellum-based cell motility. PMID- 11864998 TI - Clinically useful monoclonal antibodies in treatment. AB - Monoclonal antibodies have been used in clinical diagnosis for many years but it is only now that these agents are being licensed for clinical treatments. This review will focus on UK licensed monoclonal antibodies highlighting their clinical benefits, limitations, and side effects. PMID- 11864999 TI - Breast cancer prognostication in the 21st century and the Nottingham prognostic index. PMID- 11865000 TI - Long term prognostic value of Nottingham histological grade and its components in early (pT1N0M0) breast carcinoma. AB - AIMS: To determine the prognostic usefulness of the Nottingham histological grade (NHG) and its components in a series of 270 patients with stage pT1N0M0 breast cancer with a median follow up of 12.5 years. METHODS: Microscopic slides were re examined and the degree of tubule formation, nuclear pleomorphism, and mitotic counts were assessed and scored according to the suggested guidelines. The association with cancer specific survival (CSS) was evaluated by univariate and multivariate analyses. RESULTS: Whereas tumour size, patient age, menopausal status, type of surgery, or adjuvant treatment were not related to prognosis, histological type (p < 0.01) and NHG (p < 0.005) were associated with CSS. When evaluating the components of NHG separately, survival was not related to the score for pleomorphism, but was significantly better in tumours with score 1 or 2 for tubule formation (p < 0.007) and in those with score 1 for mitotic counts (p < 0.006). The two components retained independent significance in multivariate analysis. When the proposed cut off points for mitotic counts were replaced by lower ones based on tertile values, the mitotic index became the strongest prognostic factor (p = 0.0001) and histological type was the only additional factor of independent prognostic significance. CONCLUSIONS: These findings confirm the prognostic value of NHG in pT1N0M0 breast carcinoma, show that the evaluation of tubule formation and mitotic rate provides independent prognostic information, and suggest that the proposed cut off points for mitotic counts may be too high for this particular group of tumours. PMID- 11865001 TI - Comparison of fine needle aspiration cytology and needle core biopsy in the diagnosis of radiologically detected abdominal lesions. AB - AIMS: To compare the sensitivity and specificity of percutaneous fine needle aspiration (FNA) cytology and needle core biopsy (NCB) in the diagnosis of suspected intra-abdominal tumours. METHODS: One hundred and forty one consecutive patients who underwent radiologically guided combined FNA/NCB of abdominal lesions over a four year period were reviewed. The diagnostic accuracy of both techniques and the value of rapid staining and assessment of cytological preparations were assessed. RESULTS: FNA cytology and NCB identified 111 of 129 (86%) and 104 of 129 (80.6%) malignant lesions, respectively; in combination, the sensitivity increased to 90.7%. The diagnostic specificity was 100% for both methods, although one case of phaeochromocytoma was misinterpreted as undifferentiated carcinoma on biopsy. More accurate tumour subtying was possible in two cases with FNA and four cases on NCB. The series included 12 benign lesions, of which 11 and nine were accurately identified on FNA and NCB, respectively. Two specific benign diagnoses (Budd-Chiari syndrome and hepatic infarct) were made only on biopsy. The use of rapid assessment cytology preparations ensured that appropriate samples were submitted for microbiology in three liver abscesses, and provided an accurate cytological diagnosis at the time of the procedure in 103 of 141 (73%) cases. None of the patients suffered biopsy related complications. CONCLUSIONS: FNA cytology is more sensitive and accurate than NCB in the diagnosis of abdominal lesions, and also offers more rapid diagnosis. However, the combination of these sampling techniques increases diagnostic sensitivity and occasionally provides more accurate classification of tumours and benign lesions. The techniques should be considered complementary in the investigation of abdominal lesions. PMID- 11865002 TI - Suprabasal p53 immunoexpression is strongly associated with high grade dysplasia and risk for malignant transformation in potentially malignant oral lesions from Northern Ireland. AB - AIMS: No good predictive marker for the malignant transformation of potentially malignant oral lesions (PMOLs) is currently available. This study re-evaluated the value of p53 immunoexpression to predict malignant transformation of PMOLs after discounting possible confounding factors. METHODS: PMOLs from 18 patients who showed progression to carcinoma, 16 of the respective carcinomas, and PMOLs from 18 matched controls were evaluated by immunohistochemistry (IHC) for p53 expression. A mouse monoclonal antibody that detects wild-type and mutant forms of human p53 was used. The p53 immunostaining pattern was also correlated with the degree of dysplasia. RESULTS: Suprabasal p53 staining was significantly associated with high grades of dysplasia (p < 0.01). The specificity and positive predictive value (PPV) for malignant transformation of suprabasal p53 staining were superior to the assessment of dysplasia, but sensitivity was inferior. All carcinomas derived from PMOLs with suprabasal p53 showed strong p53 immunostaining. However, the absence of suprabasal p53 staining and/or dysplastic changes did not preclude malignant transformation in a considerable proportion of PMOLs. CONCLUSIONS: This study confirms and extends previous findings that suprabasal p53 immunoexpression has a high PPV for malignant transformation of PMOLs and can be used as a specific marker for lesions that are at high risk for malignant transformation. The absence of suprabasal p53 staining (that is, absence of, or basal, p53 staining) is non-informative for prognostic purposes. Because of its limited sensitivity, p53 IHC is not a substitute for the assessment of dysplasia in the evaluation of PMOLs. Instead, p53 IHC emerges as a clinically useful supplement of histopathological assessment in the prognosis of PMOLs. PMID- 11865003 TI - Changes in blood gas samples produced by a pneumatic tube system. AB - AIMS: To investigate the effect of a pneumatic tube system (PTS) on the results of samples sent for blood gas analysis to a central laboratory. METHODS: Blood gas samples were analysed immediately or sent via the PTS to the laboratory for analysis. In addition, samples sent via the PTS in a pressure sealed container were compared with those sent non-pressure sealed to the laboratory. RESULTS: Samples sent via the PTS had significant alterations in their pO(2) values, which were not seen when samples were carried by hand to the laboratory. There was no effect on pCO(2) and pH values. The use of a pressure sealed container abolished the alteration in pO(2) values seen. CONCLUSIONS: Samples for blood gas analysis should be transported via a PTS using a pressure sealed container to avoid artefacts in the pO(2). PMID- 11865004 TI - Detection of human papillomavirus in large cell neuroendocrine carcinoma of the uterine cervix: a study of 12 cases. AB - AIM: To investigate the role of human papillomavirus (HPV) in large cell neuroendocrine carcinoma (LCNEC) of the uterine cervix. METHODS: Twelve archival, immunohistochemically and/or electron microscopically confirmed cases of cervical LCNEC were studied. Non-isotopic in situ hybridisation (NISH) was performed on the formalin fixed, paraffin wax embedded biopsies using digoxigenin labelled probes to HPV types 6, 11, 16, 18, 31, and 33. The tumours were then subjected to polymerase chain reaction (PCR) analysis using GP5+/GP6+ consensus primers to the HPV L1 gene, in addition to type specific primers to the E6 and E6/E7 genes. RESULTS: HPV-16 was detected by NISH and/or PCR in seven of the 12 carcinomas. Two additional tumours were HPV-18 positive by NISH and/or PCR. HPV DNA was not detected in the three remaining cases. CONCLUSION: Integration of high risk HPV, in particular type 16 and to a lesser extent type 18, is associated with this uncommon variant of cervical carcinoma. PMID- 11865005 TI - Seroepidemiology of Chlamydia pneumoniae in Japan between 1991 and 2000. AB - AIM: To clarify the endemic and epidemic status of Chlamydia pneumoniae in Japan, the incidence of anti-C pneumoniae antibody was evaluated over a period of 10 years. METHOD: Serum samples were collected from 4756 healthy individuals aged 6 months to 88 years (2488 male and 2268 female individuals) between 1991 and 2000. The antibody titre was determined by a microimmunofluorescence test. RESULTS: After stratification by age and sex in each year, distinct peaks with prevalences of 73.3% and 73.0% were noted in 1993 and 1999, respectively. The lowest prevalence rate was seen in 1996 (59.0%). The epidemic cycle has been estimated to be almost six years in this geographical area. CONCLUSIONS: Chlamydia pneumoniae infection is highly endemic in Japan, as it is in Western countries, and there is a year to year variability. Long term studies in Japan are needed to clarify the epidemic occurrence of C pneumoniae infection. PMID- 11865006 TI - Her-2/neu oncogene amplification in clinically localised prostate cancer. AB - AIM: To examine the incidence of Her-2/neu oncogene amplification in clinically localised prostate cancer using in situ hybridisation. METHODS: One hundred and seventeen patients, who had undergone radical prostatectomy, were identified and in situ hybridisation was performed on formalin fixed, paraffin wax embedded tissue using the Quantum Appligene probe for Her-2/neu. The enzyme peroxidase was used to detect the probe because this enabled a permanent record to be kept. Tumours in which there were five or more signals in each nucleus in > 20% of the tumour cells were considered to have a significantly increased copy number. A serial section from these tumours was then hybridised with the chromosome 17 alpha satellite probe. The ratio of the percentage of cells showing an increase in Her-2/neu copy number to the number showing polysomy for chromosome 17 was calculated. A ratio above 2 was considered amplified. RESULTS: Biochemical recurrence occurred in 50 (43%) patients and 24 (21%) had clinical recurrence. In situ hybridisation for Her-2/neu was accessible in 114 (97%) patients. A significant increase in copy number was present in two patients (1.75 %), but chromosome 17 hybridisation showed that the increase was the result of polysomy rather than true amplification. Both these patients had a Gleason score of 7 and stage T3; they also had recurrent clinical disease with distal metastasis within two and 19 months. CONCLUSIONS: Increased Her-2/neu oncogene copy number appears to be rare in clinically localised prostatic adenocarcinoma and is related to chromosome 17 polysomy rather than true amplification. As a result, it would not be a useful biomarker for identifying those patients who will have recurrences after radical prostatectomy. PMID- 11865007 TI - Correlation between cathepsin D expression and p53 protein nuclear accumulation in oesophageal squamous cell carcinoma. AB - AIM: The lysosomal protease cathepsin D has been reported to be associated with tumour progression in malignant tumours. Expression of the gene encoding cathepsin D is known to be stimulated by oestrogen in mammary cancer cells. Recent experiments revealed that a p53 DNA binding site is located in the promoter region of the cathepsin D gene. This fact indicates that cathepsin D expression may correlate with p53 protein expression. The purpose of this study is to evaluate the expression patterns of the cathepsin D and p53 proteins in oesophageal squamous cell carcinoma (SCC). METHODS: In 154 patients with oesophageal SCC, expression of the cathepsin D and p53 proteins was measured in tumours by means of immunohistochemistry using monoclonal antibodies against cathepsin D (clone, 1C11) and p53 (clone, BP53-12). RESULTS: Cathepsin D was detected in tumour cells, although it was not found in normal oesophageal epithelium adjacent to carcinoma. High cathepsin D expression (positive tumour cells > 10%) was detected in 76 of 154 cases (49%) and high p53 nuclear expression (positive tumour cells > 50%) was detected in 70 cases (46%). High cathepsin D expression was significantly associated with invasive tumour growth (p = 0.002), poor prognosis (p = 0.049), and nuclear accumulation of p53 protein (p = 0.001). Overexpression of both p53 and cathepsin D was seen in 45 of the 154 cases (29.2%). In addition, there was a positive correlation between the cathepsin D index (percentage of cathepsin D positive tumour cells) and Ki-67 labelling index (percentage of Ki-67 positive tumour cells) in 154 oesophageal SCCs (rho = 0.257; p = 0.009). However, in multivariate survival analysis, cathepsin D expression by the tumours was not an independent prognostic factor in patients with oesophageal SCC (p = 0.236). CONCLUSIONS: The expression of cathepsin D by cancer cells may play an important role in the invasive growth of oesophageal SCC. Overexpression of both p53 and cathepsin D was seen frequently in tumours; p53 gene abnormalities may correlate with cathepsin D overexpression in oesophageal SCC. PMID- 11865008 TI - The detection of apoptosis in a human in vitro skin explant assay for graft versus host reactions. AB - AIMS: Keratinocyte apoptosis is a major pathogenic mechanism in dermal complications, such as graft versus host disease (GVHD), after allogeneic bone marrow transplantation. However, the mechanisms by which recipient target cells undergo apoptosis in GVHD are still unclear, but may result from DNA damage caused by chemotherapeutic agents and/or by direct cytokine action. The basis of this investigation was to correlate keratinocyte apoptosis with (1) the severity of graft versus host reactions (GVHR) in vitro and (2) the clinical grade (0- III) of GVHD. METHODS: Skin sections generated from an in vitro skin explant model for detecting experimental or clinically relevant GVHR were investigated for the detection of apoptotic nuclei using the terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL) technique. This investigation also aimed to establish whether the TUNEL assay could be used as an additional, predictive method for the severity of GVHD before transplantation in potential patient/donor pairs given standard GVHD prophylaxis (cyclosporin A and methotrexate). RESULTS: By comparing mean values of apoptosis for each GVHR grade in a cohort of 83 retrospective skin sections it was shown that as the severity of GVHR increased there was a parallel increase in the percentage of apoptotic cells (p < 0.0001). However, the correlation between clinical GVHD grade II--III and overall keratinocyte apoptosis (> 2.6%) did not reach this degree of significance (chi(2): 4.2; degrees of freedom, 1; p = 0.04; Fisher's exact test: p = 0.06). CONCLUSIONS: The detection of apoptosis correlated with degree of GVHR using an in vitro assay and a higher degree of apoptosis tended to correlate with more severe GVHD. Further studies in a larger cohort of patients, using other methods to detect apoptosis in conjunction with the TUNEL assay, may give additional insight into the complex immunopathophysiology of GVHD. PMID- 11865009 TI - Helicobacter pylori associated gastric diseases and lymphoid tissue hyperplasia in gastric antral mucosa. AB - AIM: To investigate the relation between Helicobacter pylori associated gastroduodenal diseases and lymphoid tissue hyperplasia in the antral mucosa and to pursue its evolution after eradication of H pylori. METHODS: Gastric antral biopsy specimens were obtained from 438 patients with H pylori positive gastroduodenal diseases (185 chronic gastritis, 69 gastric ulcer, and 184 duodenal ulcer) and 50 H pylori negative healthy controls. Lymphoid follicles and aggregates were counted and other pathological features were scored according to the updated Sydney system for classification of chronic gastritis. After a course of anti-H pylori treatment, biopsy specimens were obtained at four to six weeks, 12 months, and 24 months in the chronic gastritis patient group. RESULTS: The total prevalence of lymphoid follicles and aggregates in the biopsies was 79.9% (350 of 438; 95% confidence intervals (CI), 0.76 to 0.84). The prevalence and density of lymphoid follicles and aggregates were significantly different in the various gastroduodenal diseases. The highest prevalence (89.9%; 95% CI, 0.83 to 0.97) and density (0.82) of lymphoid follicles and aggregates occurred in patients with gastric ulcers. The lowest prevalence of lymphoid follicles and aggregates was found in patients with chronic gastritis (74.6%; 95% CI, 0.68 to 0.81), and the lowest density of lymphoid follicles and aggregates (0.56) was seen in patients with duodenal ulcers. The prevalence and density of lymphoid follicles and aggregates correlated strongly with the activity and severity of gastric antral mucosal inflammation. The eradication of H pylori resulted in a decrease in the prevalence and density of lymphoid follicles and aggregates. CONCLUSION: The prevalence and density of lymphoid follicles and aggregates in gastric antral mucosal biopsies correlated closely with H pylori infection. PMID- 11865011 TI - Clostridium novyi causing necrotising fasciitis in an injecting drug user. AB - Necrotising fasciitis with pronounced local oedema is described in an injecting drug user. Clostridium novyi was an unexpected single pathogen isolated from infected tissue. The patient was among a cluster of cases, all injecting drug users, presenting with toxaemia and soft tissue infection. The causal role and pathogenicity of C novyi is discussed. PMID- 11865010 TI - Cryptal lymphocytic coloproctitis: a new phenotype of lymphocytic colitis? AB - BACKGROUND/AIMS: Lymphocytic colitis is a clinicopathological entity characterised by protracted watery diarrhoea and an increased number of intraepithelial lymphocytes (IELs) in the surface epithelium of the colonic mucosa. This report describes two patients with symptoms similar to those of lymphocytic colitis and an increased number of IELs, but within the cryptal epithelium. METHODS: The numbers of IELs were assessed in colorectal biopsies from the two patients. Sections were stained immunohistochemically for CD3, CD8, CD20, and TIA1. RESULTS: The colorectal biopsies had an abnormally high number of IELs in the epithelium of the crypts but not in the surface epithelium. The IELs in the crypts were CD3+++, CD8+, TIA1+, and CD20-. CONCLUSIONS: The histological diagnosis in these two patients was cryptal lymphocytic coloproctitis. Patients with similar symptoms and an increased number of IELs in the surface epithelium are now filed at this department as having surface lymphocytic coloproctitis. Immunohistochemistry showed that the cryptal IELs were cytotoxic suppressor T cells. Interestingly, a case of cryptal lymphocytic colitis was recently recorded in a non-human primate dying after years of protracted chronic diarrhoea. It is possible that antigens present in the lumen of the crypts elicit a lymphocytic reaction within the cryptal cells. PMID- 11865012 TI - Massive hepatosplenomegaly caused by Penicillium marneffei associated with human immunodeficiency virus infection in a Thai patient. AB - A 29 year old Thai woman presented with non-specific features and examination revealed left upper zone consolidation and hepatosplenomegaly. The initial clinical differential diagnosis included tuberculosis and melioidosis. She died four days after admission, while still under investigation. Postmortem examination revealed antibodies to human immunodeficiency virus, disseminated Penicillium marneffei infection, and Salmonella enteritidis infection. Penicillium marneffei is a well described AIDS defining pathogen in South East Asia but is very rare in the UK. Appropriate antifungal treatment may be associated with a successful outcome. Increased awareness of this clinical association may enable correct diagnosis in affected patients from South East Asia presenting within the UK. PMID- 11865013 TI - A multicentre evaluation of the diagnostic efficiency of serological investigations for C1 inhibitor deficiency. AB - AIM: To determine the diagnostic efficiency of assays routinely used in the investigation of hereditary angio-oedema. METHODS: Over a four year period, 1144 samples were received for analysis from 907 patients suspected of C1 inhibitor deficiency. Analyses were performed for C4 and C1 inhibitor (functional and immunochemical). Notes were reviewed retrospectively on patients with low serological indicators to determine diagnosis. RESULTS: These are the first data to indicate the sensitivity, specificity, and predictive values of the assays most frequently used to screen for C1 inhibitor deficiency. A combination of low C4 and low C1 inhibitor function has 98% specificity for C1 inhibitor deficiency in this population and a 96% negative predictive value, and is thus a very effective screen. All patients with untreated C1 inhibitor deficiency had a low C4 value. CONCLUSIONS: All patients considered for a diagnosis of C1 inhibitor deficiency should have serum examined to measure both C4 and functional C1 inhibitor. If either is normal at presentation this essentially excludes a diagnosis of C1 inhibitor deficiency. These tests can be performed sequentially. If C4 is normal it is not necessary to proceed to C1 inhibitor analysis. If C1 inhibitor function and C4 are both low then a repeat sample should be obtained to confirm the findings. PMID- 11865014 TI - Progression on metastatic neuroendocrine carcinoma from a recurrent prolactinoma: a case report. AB - A 54 year old man was referred to the department of neurosurgery for frontal headache and vomiting. The patient was known in the department because of previous multiple surgery for a locally invasive pituitary prolactinoma (eight years, three years, and one year previously). The neurological examination revealed a frontal mass, which adhered to the dura, suggesting a meningioma. One year later, a left temporal metastasis was removed. Three months later, the patient died, with spinal metastases, of massive lung embolism. Histology revealed a progression of adenohypophyseal prolactinoma on neuroendocrine carcinoma, with an increase in proliferating indexes and modification of hormone production. This study documents a 10 year history of a rare prolactin producing pituitary carcinoma, which metastasised via liquoral flow. PMID- 11865015 TI - Localisation of C reactive protein in infarcted tissue sites of multiple organs during sepsis. AB - This report hypothesises an active role for the acute phase protein, C reactive protein (CRP), in local inflammatory reactions. This was studied in infarction sites from liver and kidney in a patient who died as a result of multiple complications after cholecystectomy. In this patient, a general acute phase protein reaction was induced, with an increase in plasma CRP. In infarction sites of kidney and liver, colocalisation of CRP and activated complement were found, whereas non-infarct sites were negative for CRP and complement. These results suggest that CRP directly participates in local inflammatory processes, possibly via complement activation, after binding of a suitable ligand. PMID- 11865016 TI - Changes in age related seroprevalence of antibody to varicella zoster virus: impact on vaccine strategy. AB - AIM: To study changes in the seroprevalence of varicella zoster virus (VZV) antibody over the past 25 years with a view to determining the target age group for any future vaccination strategy. METHODS: Stored sera collected from different age groups over a period of 25 years were tested by a commercial VZV IgG enzyme immunoassay at a four year time interval. Data were analysed by logistic regression to investigate the evidence for changes in incidence and hence seroprevalence over that period. RESULTS: There was a significant rise in VZV antibody prevalence in the 1-4 year age group during the study period. CONCLUSIONS: A universal childhood VZV vaccination strategy will need to take account of the increase in incidence of VZV infection in children under the age of 4 years; hence, the suggested target age would be between 12 and 18 months-- soon after the disappearance of maternal antibody. PMID- 11865017 TI - Pulmonary mucinous cystic tumour of borderline malignancy: a rare variant of adenocarcinoma. PMID- 11865018 TI - Renal oncocytoma with a novel chromosomal rearrangement, der(13)t(13;16)(p11;p11), associated with a renal cell carcinoma. PMID- 11865019 TI - Differential regulation of ER Ca2+ uptake and release rates accounts for multiple modes of Ca2+-induced Ca2+ release. AB - The ER is a central element in Ca(2+) signaling, both as a modulator of cytoplasmic Ca(2+) concentration ([Ca(2+)](i)) and as a locus of Ca(2+)-regulated events. During surface membrane depolarization in excitable cells, the ER may either accumulate or release net Ca(2+), but the conditions of stimulation that determine which form of net Ca(2+) transport occurs are not well understood. The direction of net ER Ca(2+) transport depends on the relative rates of Ca(2+) uptake and release via distinct pathways that are differentially regulated by Ca(2+), so we investigated these rates and their sensitivity to Ca(2+) using sympathetic neurons as model cells. The rate of Ca(2+) uptake by SERCAs (J(SERCA)), measured as the t-BuBHQ-sensitive component of the total cytoplasmic Ca(2+) flux, increased monotonically with [Ca(2+)](i). Measurement of the rate of Ca(2+) release (J(Release)) during t-BuBHQ-induced [Ca(2+)](i) transients made it possible to characterize the Ca(2+) permeability of the ER ((~)P(ER)), describing the activity of all Ca(2+)-permeable channels that contribute to passive ER Ca(2+) release, including ryanodine-sensitive Ca(2+) release channels (RyRs) that are responsible for CICR. Simulations based on experimentally determined descriptions of J(SERCA), and of Ca(2+) extrusion across the plasma membrane (J(pm)) accounted for our previous finding that during weak depolarization, the ER accumulates Ca(2+), but at a rate that is attenuated by activation of a CICR pathway operating in parallel with SERCAs to regulate net ER Ca(2+) transport. Caffeine greatly increased the [Ca(2+)] sensitivity of ((~)P(ER)), accounting for the effects of caffeine on depolarization-evoked [Ca(2+)](i) elevations and caffeine-induced [Ca(2+)](i) oscillations. Extending the rate descriptions of J(SERCA), ((~)P(ER)), and J(pm) to higher [Ca(2+)](i) levels shows how the interplay between Ca(2+) transport systems with different Ca(2+) sensitivities accounts for the different modes of CICR over different ranges of [Ca(2+)](i) during stimulation. PMID- 11865020 TI - Single ion occupancy and steady-state gating of Na channels in squid giant axon. AB - The properties of the small fraction of tetrodotoxin (TTX)-sensitive Na channels that remain open in the steady state were studied in internally dialyzed voltage clamped squid giant axons. The observed Ussing flux ratio exponent (n') of 0.97 plus minus 0.03 (calculated from simultaneous measurements of TTX-sensitive current and (22)Na efflux) and nonindependent behavior of Na current at high internal [Na] are explained by a one-site ("1s") permeation model characterized by a single effective binding site within the channel pore in equilibrium with internal Na ions (apparent equilibrium dissociation constant K(Nai)(0) = 0.61 +/- 0.08 M). Steady-state open probability of the TTX-sensitive channels can be modeled by the product p(a)p(infinity), where p(a) represents voltage-dependent activation described by a Boltzmann distribution with midpoint V(a) = -7 mV and effective valence z(a) = 3.2 (Vandenberg, C.A., and F. Bezanilla. 1991. BIOPHYS: J. 60:1499--1510) coupled to voltage-independent inactivation by an equilibrium constant (Bezanilla, F., and C.M. Armstrong. 1977. J. Gen. Physiol. 70:549--566) K(eq) = 770. The factor p(infinity) represents voltage-dependent inactivation with empirical midpoint V(infinity)= -83 plus minus 5 mV and effective valence z(infinity) = 0.55 plus minus 0.03. The composite p(a)p(infinity)1s model describes the steady-state voltage dependence of the persistent TTX-sensitive current well. PMID- 11865021 TI - Natural bile acids and synthetic analogues modulate large conductance Ca2+ activated K+ (BKCa) channel activity in smooth muscle cells. AB - Bile acids have been reported to produce relaxation of smooth muscle both in vitro and in vivo. The cellular mechanisms underlying bile acid-induced relaxation are largely unknown. Here we demonstrate, using patch-clamp techniques, that natural bile acids and synthetic analogues reversibly increase BK(Ca) channel activity in rabbit mesenteric artery smooth muscle cells. In excised inside-out patches bile acid-induced increases in channel activity are characterized by a parallel leftward shift in the activity-voltage relationship. This increase in BK(Ca) channel activity is not due to Ca(2+)-dependent mechanism(s) or changes in freely diffusible messengers, but to a direct action of the bile acid on the channel protein itself or some closely associated component in the cell membrane. For naturally occurring bile acids, the magnitude of bile acid-induced increase in BK(Ca) channel activity is inversely related to the number of hydroxyl groups in the bile acid molecule. By using synthetic analogues, we demonstrate that such increase in activity is not affected by several chemical modifications in the lateral chain of the molecule, but is markedly favored by polar groups in the side of the steroid rings opposite to the side where the methyl groups are located, which stresses the importance of the planar polarity of the molecule. Bile acid-induced increases in BK(Ca) channel activity are also observed in smooth muscle cells freshly dissociated from rabbit main pulmonary artery and gallbladder, raising the possibility that a direct activation of BK(Ca) channels by these planar steroids is a widespread phenomenon in many smooth muscle cell types. Bile acid concentrations that increase BK(Ca) channel activity in mesenteric artery smooth muscle cells are found in the systemic circulation under a variety of human pathophysiological conditions, and their ability to enhance BK(Ca) channel activity may explain their relaxing effect on smooth muscle. PMID- 11865022 TI - Fast and slow voltage sensor movements in HERG potassium channels. AB - HERG encodes an inwardly-rectifying potassium channel that plays an important role in repolarization of the cardiac action potential. Inward rectification of HERG channels results from rapid and voltage-dependent inactivation gating, combined with very slow activation gating. We asked whether the voltage sensor is implicated in the unusual properties of HERG gating: does the voltage sensor move slowly to account for slow activation and deactivation, or could the voltage sensor move rapidly to account for the rapid kinetics and intrinsic voltage dependence of inactivation? To probe voltage sensor movement, we used a fluorescence technique to examine conformational changes near the positively charged S4 region. Fluorescent probes attached to three different residues on the NH(2)-terminal end of the S4 region (E518C, E519C, and L520C) reported both fast and slow voltage-dependent changes in fluorescence. The slow changes in fluorescence correlated strongly with activation gating, suggesting that the slow activation gating of HERG results from slow voltage sensor movement. The fast changes in fluorescence showed voltage dependence and kinetics similar to inactivation gating, though these fluorescence signals were not affected by external tetraethylammonium blockade or mutations that alter inactivation. A working model with two types of voltage sensor movement is proposed as a framework for understanding HERG channel gating and the fluorescence signals. PMID- 11865024 TI - All TRAFs are not created equal: common and distinct molecular mechanisms of TRAF mediated signal transduction. AB - The tumor necrosis factor (TNF) receptor associated factors (TRAFs) have emerged as the major signal transducers for the TNF receptor superfamily and the interleukin-1 receptor/Toll-like receptor (IL-1R/TLR) superfamily. TRAFs collectively play important functions in both adaptive and innate immunity. Recent functional and structural studies have revealed the individuality of each of the mammalian TRAFs and advanced our understanding of the underlying molecular mechanisms. Here, we examine this functional divergence among TRAFs from a perspective of both upstream and downstream TRAF signal transduction pathways and of signaling-dependent regulation of TRAF trafficking. We raise additional questions and propose hypotheses regarding the molecular basis of TRAF signaling specificity. PMID- 11865025 TI - Biological roles and mechanistic actions of co-repressor complexes. AB - Transcriptional repression, which plays a crucial role in diverse biological processes, is mediated in part by non-DNA-binding co-repressors. The closely related co-repressor proteins N-CoR and SMRT, although originally identified on the basis of their ability to associate with and confer transcriptional repression through nuclear receptors, have been shown to be recruited to many classes of transcription factor and are in fact components of multiple protein complexes containing histone deacetylase proteins. This association with histone deacetylase activity provides an important component of the mechanism that allows DNA-binding proteins interacting with N-CoR or SMRT to repress transcription of specific target genes. Both N-CoR and SMRT are important targets for cell signaling pathways, which influence their expression levels, subcellular localization and association with other proteins. Recently, the biological importance of these proteins has been revealed by studies of genetically engineered mice and human diseases such as acute promyelocytic leukemia (APL) and resistance to thyroid hormone (RTH). PMID- 11865026 TI - N-WASP activation by a beta1-integrin-dependent mechanism supports PI3K independent chemotaxis stimulated by urokinase-type plasminogen activator. AB - Urokinase-type plasminogen activator (uPA)-uPA receptor (uPAR) and epidermal growth factor (EGF)-EGF receptor (EGFR) expression is highly correlated with breast cancer metastasis. Phosphoinositide 3-kinase (PI3K), small Rho GTPases, such as Cdc42 and Rac1, and neuronal Wiskott Aldrich syndrome protein (N-WASP) are key effectors that regulate dynamic changes in the actin cytoskeleton and cell migration. uPA- and EGF-stimulated chemotaxis, cytoskeletal rearrangements and activation of Cdc42, Rac1 and N-WASP were studied in the highly metastatic human breast cancer cell line MDA MB 231. These studies reveal that divergent signalling occurs downstream of PI3K. The activity of PI3K was not necessary for uPA-induced chemotactic responses, but those induced by EGF were entirely dependent upon PI3K. Furthermore, PI3K-independent chemotactic signalling by uPA was shown to involve disruption of an interaction between beta(1)-integrins and N WASP and translocation of N-WASP to the actin cytoskeleton. PMID- 11865027 TI - Vimentin affects localization and activity of sodium-glucose cotransporter SGLT1 in membrane rafts. AB - It has been reported that vimentin, a cytoskeleton filament that is expressed only in mesenchymal cells after birth, is re-expressed in epithelial cells in vivo under pathological conditions and in vitro in primary culture. Whether vimentin re-expression is only a marker of cellular dedifferentiation or is instrumental in the maintenance of cell structure and/or function is a matter of debate. To address this issue, we used renal proximal tubular cells in primary culture from vimentin-null mice (Vim(-/-)) and from wild-type littermates (Vim(+/+)). The absence of vimentin did not affect cell morphology, proliferation and activity of hydrolases, but dramatically decreased Na-glucose cotransport activity. This phenotype was associated with a specific reduction of SGLT1 protein in the detergent-resistant membrane microdomains (DRM). In Vim(+/+) cells, disruption of these microdomains by methyl-beta-cyclodextrin decreased SGLT1 protein abundance in DRM, a change that was paralleled by a decrease of Na glucose transport activity. Importantly, we showed that vimentin is located to DRM, but it disappeared after methyl-beta-cyclodextrin treatment. In Vim(-/-) cells, supplementation of cholesterol with cholesterol-methyl-beta-cyclodextrin complexes completely restored Na-glucose transport activity. Interestingly, neither cholesterol content nor cholesterol metabolism changed in Vim(-/-) cells. Our results are consistent with the view that re-expression of vimentin in epithelial cells could be instrumental to maintain the physical state of rafts and, thus, the function of DRM-associated proteins. PMID- 11865028 TI - Domains of the Pavarotti kinesin-like protein that direct its subcellular distribution: effects of mislocalisation on the tubulin and actin cytoskeleton during Drosophila oogenesis. AB - The kinesin-like protein encoded by pavarotti (Pav-KLP) is essential for cytokinesis and associates with the central part of the late mitotic spindle and interphase nuclei in somatic cells (Adams et al., 1988). Here we define regions of the molecule that regulate its subcellular localisation and study the consequences of overexpressing mutant forms of the protein during oogenesis in Drosophila. Pav-KLP normally associates with the oocyte nucleus, but when over expressed at moderate levels, its GFP tagged form also accumulates in nurse cell nuclei. At high expression levels this leads to loss of the microfilaments that tether these nuclei, so that they block the ring canals and prevent the 'dumping' of nurse cell cytoplasm into the oocyte, which results in sterility. Localisation to these nuclei is prevented by mutations in either the conserved ATP-binding site of the motor domain or the nuclear localisation sequences in the C-terminal domain. Both such mutations lead to the formation of stable arrays of cytoplasmic microtubules and the progressive disruption of the actin cytoskeleton. The latter is evident by a breakdown of the cortical actin causing disruption of cell membranes; this breakdown ultimately results in the accumulation of cytoplasmic aggregates containing tubulin, actin and at least some of their binding proteins. Pav-KLP is also found associated with the ring canals, actin-rich structures built from remnants of the cytokinesis ring. The stalk domain alone is sufficient for the exclusive association of Pav-KLP to these structures, and this has no consequences for fertility. We discuss whether disruption of actin structures by full-length cytoplasmic forms of Pav-KLP is a consequence of the resulting stabilised cytoplasmic microtubules per se or accumulation of the motor protein at ectopic cortical sites to sequester molecules that regulate actin behaviour. PMID- 11865029 TI - Two members of the beige/CHS (BEACH) family are involved at different stages in the organization of the endocytic pathway in Dictyostelium. AB - Proteins of the Chediak-Higashi/Beige (BEACH) family have been implicated in the function of lysosomes, as well as in signal transduction, but their molecular role is still poorly understood. In Dictyostelium, at least six members of the family can be identified. Here cells with mutations in two of these genes, LVSA and LVSB, were analyzed. Interestingly both mutants exhibited defects in the organization of the endocytic pathway, albeit at distinct stages. In lvsB mutant cells, the regulated secretion of lysosomal enzymes was enhanced, a phenotype reminiscent of the Chediak-Higashi syndrome. LvsA mutant cells exhibited alterations in the organization and function of the early endocytic and phagocytic pathway. The LvsA protein may participate in the signaling pathway, which links adhesion of a particle to the subsequent formation of a phagocytic cup. Further genetic analysis will be necessary to determine whether other members of the BEACH family of proteins are also involved in controlling the organization of the endocytic pathway. PMID- 11865030 TI - Poly-gamma-glutamate synthesis during formation of nematocyst capsules in Hydra. AB - Nematocysts are explosive organelles found in all Cnidaria. Explosion of nematocyst capsules is driven by the high pressure within the capsule formed by the high concentration of poly-gamma-glutamate in the capsule matrix. Poly-gamma glutamate is a polyanion that binds cations tightly, including the fluorescent cationic dyes acridine orange and DAPI (4',6-diamidino-2-phenylindole). We have used acridine orange and DAPI staining to localize poly-gamma-glutamate within capsules and to follow the biosynthesis of poly-gamma-glutamate during capsule formation. The results indicate that poly-gamma-glutamate biosynthesis occurs late in capsule formation after invagination of the tubule and that it is accompanied by swelling of the capsule due to increasing osmotic pressure. The matrix in all four capsule types is homogeneously filled with poly-gamma glutamate. In vivo this poly-gamma-glutamate is complexed with monovalent cations. In addition, poly-gamma-glutamate is formed within the tubule lumen of stenoteles. We argue that this poly-gamma-glutamate is required to drive the two step explosion process in stenotele nematocysts. PMID- 11865031 TI - Bone morphogenetic proteins promote development of fetal pancreas epithelial colonies containing insulin-positive cells. AB - Extracellular signals that guide pancreas cell development are not well characterized. In an in vitro culture system of dissociated pancreas cells from the E15.5 mouse fetus we show that, in the presence of the extracellular matrix protein laminin-1, bone morphogenetic proteins (BMPs-4, -5 and -6) promote the development of cystic epithelial colonies. Transforming growth factor beta1 (TGF beta1) and activin A antagonise this effect of BMP-6 and inhibit colony formation. Histological analysis revealed that the colonies are composed of E cadherin-positive epithelial cells, which in localised areas are insulin positive. The colonies also contain occasional glucagon-positive cells, but no somatostatin- or alpha-amylase-positive cells. These findings indicate that members of the TGF-beta superfamily regulate pancreas epithelial cell development and can promote the formation of islet-like structures in vitro. PMID- 11865032 TI - P35-sensitive caspases, MAP kinases and Rho modulate beta-adrenergic induction of apoptosis in mollusc immune cells. AB - Apoptosis is an important mechanism for the preservation of a healthy and balanced immune system in vertebrates. Little is known, however, about how apoptotic processes regulate invertebrate immune defenses. In the present study, we show that noradrenaline, a catecholamine produced by the neuroendocrine system and by immune cells in molluscs, is able to induce apoptosis of oyster Crassostrea gigas hemocytes. The apoptosis-inducing effect of noradrenaline was mimicked by isoproterenol and blocked by propranolol, which indicates that noradrenaline triggers apoptosis via a beta-adrenergic signaling pathway. Exposure to the pan-caspase inhibitor Z-VAD-FMK or expression of the caspase inhibitor P35 under the transcriptional control of a mollusc hsp70 gene promoter reduced the number of apoptotic cells among noradrenaline-treated hemocytes. These results suggest that P35-sensitive caspases are involved in the apoptotic process triggered by beta-adrenergic signaling. Complementary experiments suggest that mitogen-activated protein kinases and Rho, a member of the Ras GTPase family, may be involved in antiapoptotic mechanisms that modulate the apoptotic effect of noradrenaline. Taken together, these results provide a first insight into apoptotic processes in mollusc immune cells. PMID- 11865033 TI - The T-box transcription factor Brachyury mediates cartilage development in mesenchymal stem cell line C3H10T1/2. AB - The BMP2-dependent onset of osteo/chondrogenic differentiation in the acknowledged pluripotent murine mesenchymal stem cell line (C3H10T1/2) is accompanied by the immediate upregulation of Fibroblast Growth Factor Receptor 3 (FGFR3) and a delayed response by FGFR2. Forced expression of FGFR3 in C3H10T1/2 is sufficient for chondrogenic differentiation, indicating an important role for FGF-signaling during the manifestation of the chondrogenic lineage in this cell line. Screening for transcription factors exhibiting a chondrogenic capacity in C3H10T1/2 identified that the T-box containing transcription factor Brachyury is upregulated by FGFR3-mediated signaling. Forced expression of Brachyury in C3H10T1/2 was sufficient for differentiation into the chondrogenic lineage in vitro and in vivo after transplantation into muscle. A dominant-negative variant of Brachyury, consisting of its DNA-binding domain (T-box), interferes with BMP2 mediated cartilage formation. These studies indicate that BMP-initiated FGF signaling induces a novel type of transcription factor for the onset of chondrogenesis in a mesenchymal stem cell line. A potential role for this T-box factor in skeletogenesis is further delineated from its expression profile in various skeletal elements such as intervertebral disks and the limb bud at late stages (18.5 d.p.c.) of murine embryonic development. PMID- 11865034 TI - Mechanisms of CFTR regulation by syntaxin 1A and PKA. AB - Activation of the chloride selective anion channel CFTR is stimulated by cAMP dependent phosphorylation and is regulated by the target membrane t-SNARE syntaxin 1A. The mechanism by which SNARE proteins modulate CFTR in secretory epithelia is controversial. In addition, controversy exists as to whether PKA activates CFTR-mediated Cl(-) currents (I(CFTR)) by increasing the number of channels in the plasma membrane and/or by stimulating membrane-resident channels. SNARE proteins play a well known role in exocytosis and have recently been implicated in the regulation of ion channels; therefore this investigation sought to resolve two related issues: (a) is PKA activation or SNARE protein modulation of CFTR linked to changes in membrane turnover and (b) does syntaxin 1A modulate CFTR via direct effects on the gating of channels residing in the plasma membrane versus alterations in membrane traffic. Our data demonstrate that syntaxin 1A inhibits CFTR as a result of direct protein-protein interactions that decrease channel open probability (P(o)) and serves as a model for other SNARE protein-ion channel interactions. We also show that PKA activation can enhance membrane trafficking in some epithelial cell types, and this is independent from CFTR activation or syntaxin 1A association. PMID- 11865035 TI - UV induces tyrosine kinase-independent internalisation and endosome arrest of the EGF receptor. AB - We have compared the activation and trafficking of epidermal growth factor receptor (EGFR) induced by UV light and EGF. Tyrosine phosphorylation of EGFR was not detected in UV-exposed cells by immunoblotting of whole cell lysates or EGFR immunoprecipitates with antibodies specific for each of the five activated autophosphorylation sites of EGFR. In addition, EGFR of UV-irradiated cells did not demonstrate increased (32)P-incorporation. However, UV-exposed cells demonstrated a gel mobility shift of EGFR, which was not abolished by alkaline phosphatase treatment. UV-exposure did not induce dimerisation of EGFR. Furthermore, UV induced internalisation of EGFR without polyubiquitination or degradation. UV-exposed EGFR was transferred to early endosomes and arrested in transferrin-accessible endosomes close to the cell surface. Whereas inhibition of the EGFR tyrosine kinase effectively inhibited tyrosine phosphorylation and internalisation of EGF-activated EGFR, internalisation of UV-exposed EGFR was unaffected. UV induced neither relocalisation of Shc and Grb2 nor activation of Raf, but activation of MEK and MAPK was observed. Our work indicates that UV induces internalisation of EGFR independent of its phosphorylation or receptor tyrosine kinase activation, and altered EGFR trafficking compared with ligand activated receptor. In addition, MAPK activation by UV does not appear to be mediated by EGFR activation. PMID- 11865036 TI - Mutational analysis of the variant surface glycoprotein GPI-anchor signal sequence in Trypanosoma brucei. AB - The variant surface glycoproteins (VSG) of Trypanosoma brucei are anchored to the cell surface via a glycosylphosphatidylinositol (GPI) anchor. All GPI-anchored proteins are synthesized with a C-terminal signal sequence, which is replaced by a GPI-anchor in a rapid post-translational transamidation reaction. VSG GPI signal sequences are extraordinarily conserved. They contain either 23 or 17 amino acids, a difference that distinguishes the two major VSG classes, and consist of a spacer sequence followed by a more hydrophobic region. The omega amino acid, to which GPI is transferred, is either Ser, Asp or Asn, the omega+2 amino acid is always Ser, and the omega+7 amino acid is almost always Lys. In order to determine whether this high conservation is necessary for GPI anchoring, we introduced several mutations into the signal peptide. Surprisingly, changing the most conserved amino acids, at positions omega+1, omega+2 and omega+7, had no detectable effect on the efficiency of GPI-anchoring or on protein abundance. Several more extensive changes also had no discernable impact on GPI-anchoring. Deleting the entire 23 amino-acid signal sequence or the 15 amino-acid hydrophobic region generated proteins that were not anchored. Instead of being secreted, these truncated proteins accumulated in the endoplasmic reticulum prior to lysosomal degradation. Replacing the GPI signal sequence with a proven cell surface membrane-spanning domain reduced expression by about 99% and resulted not in cell surface expression but in accumulation close to the flagellar pocket and in non-lysosomal compartments. These results indicate that the high conservation of the VSG GPI signal sequence is not necessary for efficient expression and GPI attachment. Instead, the GPI anchor is essential for surface expression of VSG. However, because the VSG is a major virulence factor, it is possible that small changes in the efficiency of GPI anchoring, undetectable in our experiments, might have influenced the evolution of VSG GPI signal sequences. PMID- 11865037 TI - Differential expression of receptors for Shiga and Cholera toxin is regulated by the cell cycle. AB - Cholera and Shiga toxin bind to the cell surface via glycolipid receptors GM1 and Gb3, respectively. Surprisingly, the majority of Vero cells from a non synchronized population bind either Cholera or Shiga toxin but not both toxins. The hypothesis that the differential expression of toxin receptors is regulated by the cell cycle was tested. We find that Cholera toxin binds preferentially in G0/G1, with little binding through S-phase to telophase, whereas Shiga toxin binds maximally through G2 to telophase but does not bind during G0/G1 and S phase. The changes result from the corresponding changes in Gb3 and GM1 synthesis, not from variations of receptor transport to the cell surface. The changes do not reflect competition of Gb3 and GM1 synthesis for lactosylceramide. Cells as diverse as Vero cells, PC12 cells and astrocytes show the same cell cycle-dependent regulation of glycosphingolipid receptors, suggesting that this novel phenomenon is based on a conserved regulatory mechanism. PMID- 11865038 TI - Identification and characterization of a novel human plant pathogenesis-related protein that localizes to lipid-enriched microdomains in the Golgi complex. AB - Group 1 of plant pathogenesis-related proteins (PR-1) and a variety of related mammalian proteins constitute a superfamily of proteins that share structural similarities. Little is known about their function, but all the family members identified to date are co-translationally translocated to the lumen of the endoplasmic reticulum and are secreted as soluble proteins or are targeted to vacuoles. Here we report the identification of a novel family member that localizes to the cytosolic site of the endomembrane system in mammalian cells. After detergent solubilization of isolated Golgi membranes, a 17 kDa protein was found associated with a low-density detergent-insoluble fraction. The amino-acid sequence, determined by microsequencing and molecular cloning, revealed a significant homology with the superfamily of PR-1 proteins. Golgi-associated PR-1 protein (GAPR-1) showed a brefeldin-A-sensitive Golgi localization in immunofluorescence. Interestingly, the protein remained associated with the microdomain fraction in the presence of Brefeldin A. By mass spectrometry, GAPR-1 was shown to be myristoylated. Immunoprecipitation of GAPR- 1 from Golgi membranes resulted in the coimmunoprecipitation of caveolin-1, indicating a direct interaction between these two proteins. Myristoylation, together with protein-protein or electrostatic interactions at physiological pH owing to the highly basic pI of GAPR-1 (pI 9.4) could explain the strong membrane association of GAPR-1. Tissue screening revealed that GAPR-1 is not detectably expressed in liver, heart or adrenal glands. High expression was found in monocytes, leukocytes, lung, spleen and embryonic tissue. Consistent with the involvement of PR-1 proteins in the plant immune system, these data could indicate that GAPR-1 is involved in the immune system. PMID- 11865040 TI - Fc-receptor-mediated phagocytosis is regulated by mechanical properties of the target. AB - Phagocytosis is an actin-based process used by macrophages to clear particles greater than 0.5 microm in diameter. In addition to its role in immunological responses, phagocytosis is also necessary for tissue remodeling and repair. To prevent catastrophic autoimmune reactions, phagocytosis must be tightly regulated. It is commonly assumed that the recognition/selection of phagocytic targets is based solely upon receptor-ligand binding. Here we report an important new criterion, that mechanical parameters of the target can dramatically affect the efficiency of phagocytosis. When presented with particles of identical chemical properties but different rigidity, macrophages showed a strong preference to engulf rigid objects. Furthermore, phagocytosis of soft particles can be stimulated with the microinjection of constitutively active Rac1 but not RhoA, and with lysophosphatidic acid, an agent known to activate the small GTP binding proteins of the Rho family. These data suggest a Rac1-dependent mechanosensory mechanism for phagocytosis, which probably plays an important role in a number of physiological and pathological processes from embryonic development to autoimmune diseases. PMID- 11865039 TI - Signaling through the EGF receptor controls lung morphogenesis in part by regulating MT1-MMP-mediated activation of gelatinase A/MMP2. AB - Epithelial-mesenchymal interactions during lung development require extracellular signaling factors that facilitate branching morphogenesis. We show here that matrix metalloproteinases (MMPs) originating in the mesenchyme are necessary for epithelial branching and alveolization. We found that the delayed lung maturation characterized by abnormal branching and poor alveolization seen in mice deficient in epidermal growth factor receptor (Egfr(-/-)) is accompanied by aberrant expression of MMPs. By in situ zymography, the lungs from newborn Egfr(-/-) mice had low gelatinolytic activity compared with wildtype. Inhibition of MMPs in developing lungs in vivo or in vitro severely retarded morphogenesis. Egfr(-/-) mice had low expression of MT1-MMP/MMP14, which is a potent activator of gelatinase A/MMP2, in their lungs. Egf ligand increased MT1-MMP mRNA by tenfold in lung fibroblasts from wild type, but not from Egfr(-/-) mice. Extracts from lungs of Egfr(-/-) mice showed a tenfold reduction in active MMP-2, but only a slight decrease in proMMP-2 by zymography. At birth, MMP-2(-/-) mice had a lung phenotype characterized by abnormal lung alveolization which phenocopied that of Egfr(-/-) mice, albeit somewhat less severe. We conclude that proteolysis mediates epithelial/mesenchymal interactions during lung morphogenesis. From the phenotypes of the Egfr(-/-) mice, we identify MT1-MMP as a major downstream target of Egfr signaling in lung in vivo and in vitro. MT1-MMP is, in turn, necessary for activation of MMP-2, a mesenchymal enzyme that is required for normal lung morphogenesis. PMID- 11865041 TI - The Caenorhabditis elegans histone hairpin-binding protein is required for core histone gene expression and is essential for embryonic and postembryonic cell division. AB - As in all metazoans, the replication-dependent histone genes of Caenorhabditis elegans lack introns and contain a short hairpin structure in the 3' untranslated region. This hairpin structure is a key element for post-transcriptional regulation of histone gene expression and determines mRNA 3' end formation, nuclear export, translation and mRNA decay. All these steps contribute to the S phase-specific expression of the replication-dependent histone genes. The hairpin structure is the binding site for histone hairpin-binding protein that is required for hairpin-dependent regulation. Here, we demonstrate that the C. elegans histone hairpin-binding protein gene is transcribed in dividing cells during embryogenesis and postembryonic development. Depletion of histone hairpin binding protein (HBP) function in early embryos using RNA-mediated interference leads to an embryonic-lethal phenotype brought about by defects in chromosome condensation. A similar phenotype was obtained by depleting histones H3 and H4 in early embryos, indicating that the defects in hairpin-binding protein-depleted embryos are caused by reduced histone biosynthesis. We have confirmed this by showing that HBP depletion reduces histone gene expression. Depletion of HBP during postembryonic development also results in defects in cell division during late larval development. In addition, we have observed defects in the specification of vulval cell fate in animals depleted for histone H3 and H4, which indicates that histone proteins are required for cell fate regulation during vulval development. PMID- 11865042 TI - Transcription activator interactions with multiple SWI/SNF subunits. AB - We have previously shown that the yeast SWI/SNF complex stimulates in vitro transcription from chromatin templates in an ATP-dependent manner. SWI/SNF function in this regard requires the presence of an activator with which it can interact directly, linking activator recruitment of SWI/SNF to transcriptional stimulation. In this study, we determine the SWI/SNF subunits that mediate its interaction with activators. Using a photo-cross-linking label transfer strategy, we show that the Snf5, Swi1, and Swi2/Snf2 subunits are contacted by the yeast acidic activators, Gcn4 and Hap4, in the context of the intact native SWI/SNF complex. In addition, we show that the same three subunits can interact individually with acidic activation domains, indicating that each subunit contributes to binding activators. Furthermore, mutations that reduce the activation potential of these activators also diminish its interaction with each of these SWI/SNF subunits. Thus, three distinct subunits of the SWI/SNF complex contribute to its interactions with activation domains. PMID- 11865043 TI - Competitive cofactor recruitment by orphan receptor hepatocyte nuclear factor 4alpha1: modulation by the F domain. AB - For most ligand-dependent nuclear receptors, the status of endogenous ligand modulates the relative affinities for corepressor and coactivator complexes. It is less clear what parameters modulate the switch between corepressor and coactivator for the orphan receptors. Our previous work demonstrated that hepatocyte nuclear factor 4alpha1 (HNF4alpha1, NR2A1) interacts with the p160 coactivator GRIP1 and the cointegrators CBP and p300 in the absence of exogenously added ligand and that removal of the F domain enhances these interactions. Here, we utilized transient-transfection analysis to demonstrate repression of HNF4alpha1 activity by the corepressor silencing mediator of retinoid and thyroid receptors (SMRT) in several cell lines and on several HNF4alpha-responsive promoter elements. Glutathione S-transferase pulldown assays confirmed a direct interaction between HNF4alpha1 and receptor interaction domain 2 of SMRT. Loss of the F domain resulted in marked reduction of the ability of SMRT to interact with HNF4alpha1 in vitro and repress HNF4alpha1 activity in vivo, although the isolated F domain itself failed to interact with SMRT. Surprisingly, loss of both the A/B and F domains restored full repression by SMRT, suggesting involvement of both domains in the SMRT interaction. Finally, we show that when coexpressed along with HNF4alpha1 and GRIP1, CBP, or p300, SMRT can titer out HNF4alpha1-mediated transactivation in a dose-dependent manner and that this competition derives from mutually exclusive binding. Collectively, these results suggest that HNF4alpha can functionally interact with both a coactivator and a corepressor without altering the status of any putative ligand and that the presence of the F domain may play a role in discriminating between the different coregulators. PMID- 11865044 TI - Peroxisomal targeting signal receptor Pex5p interacts with cargoes and import machinery components in a spatiotemporally differentiated manner: conserved Pex5p WXXXF/Y motifs are critical for matrix protein import. AB - Two isoforms of the peroxisomal targeting signal type 1 (PTS1) receptor, termed Pex5pS and (37-amino-acid-longer) Pex5pL, are expressed in mammals. Pex5pL transports PTS1 proteins and Pex7p-PTS2 cargo complexes to the initial Pex5p docking site, Pex14p, on peroxisome membranes, while Pex5pS translocates only PTS1 cargoes. Here we report functional Pex5p domains responsible for interaction with peroxins Pex7p, Pex13p, and Pex14p. An N-terminal half, such as Pex5pL(1 243), comprising amino acid residues 1 to 243, bound to Pex7p, Pex13p, and Pex14p and was sufficient for restoring the impaired PTS2 import of pex5 cell mutants, while the C-terminal tetratricopeptide repeat motifs were required for PTS1 binding. N-terminal Pex5p possessed multiple Pex14p-binding sites. Alanine scanning analysis of the highly conserved seven (six in Pex5pS) pentapeptide WXXXF/Y motifs residing at the N-terminal region indicated that these motifs were essential for the interaction of Pex5p with Pex14p and Pex13p. Moreover, mutation of several WXXXF/Y motifs did not affect the PTS import-restoring activity of Pex5p, implying that the binding of Pex14p to all of the WXXXF/Y sites was not a prerequisite for the translocation of Pex5p-cargo complexes. Pex5p bound to Pex13p at the N-terminal part, not to the C-terminal SH3 region, via WXXXF/Y motifs 2 to 4. PTS1 and PTS2 import required the interaction of Pex5p with Pex14p but not with Pex13p, while Pex5p binding to Pex13p was essential for import of catalase with PTS1-like signal KANL. Pex5p recruited PTS1 proteins to Pex14p but not to Pex13p. Pex14p and Pex13p formed a complex with PTS1-loaded Pex5p but dissociated in the presence of cargo-unloaded Pex5p, implying that PTS cargoes are released from Pex5p at a step downstream of Pex14p and upstream of Pex13p. Thus, Pex14p and Pex13p very likely form mutually and temporally distinct subcomplexes involved in peroxisomal matrix protein import. PMID- 11865046 TI - RNA destabilization by the granulocyte colony-stimulating factor stem-loop destabilizing element involves a single stem-loop that promotes deadenylation. AB - Granulocyte colony-stimulating factor (G-CSF) mRNA contains two distinct types of cis-acting mRNA destabilizing elements in the 3'-untranslated region. In addition to several copies of the AU-rich element the G-CSF mRNA also contains a destabilizing region that includes several predicted stem-loop structures. We report here that the destabilizing activity resides in a single stem-loop structure within this region. A consensus sequence for the active structure has been derived by site-directed mutagenesis, revealing that a three-base loop of sequence YAU and unpaired bases either side of the stem contribute to the activity. The helical nature of the stem is essential and the stem must be less than 11 bp in length, but the destabilizing activity is relatively insensitive to the sequence within the helix. The stem-loop increases the rate of mRNA deadenylation, most likely by enhancing the processivity of the deadenylation reaction. A protein that binds the stem-loop, but not an inactive mutant form, has been detected in cytoplasmic lysates. PMID- 11865045 TI - Phosphorylation of eukaryotic translation initiation factor 4E is critical for growth. AB - Eukaryotic translation initiation factor 4E (eIF4E) binds to the cap structure at the 5' end of mRNAs and is a critical target for the control of protein synthesis. eIF4E is phosphorylated in many systems in response to extracellular stimuli, but biochemical evidence to date has been equivocal as to the biological significance of this modification. Here we use a genetic approach to this problem. We show that, in Drosophila melanogaster, homozygous eIF4E mutants arrest growth during larval development. In Drosophila eIF4EI, Ser251 corresponds to Ser209 of mammalian eIF4E, which is phosphorylated in response to extracellular signals. We find that, in vivo, eIF4EI Ser251 mutants cannot incorporate labeled phosphate. Furthermore, transgenic Drosophila organisms expressing eIF4E(Ser251Ala) in an eIF4E mutant background have reduced viability. Escapers develop more slowly than control siblings and are smaller. These genetic data provide evidence that eIF4E phosphorylation is biologically significant and is essential for normal growth and development. PMID- 11865047 TI - Caspase cleavage of initiation factor 4E-binding protein 1 yields a dominant inhibitor of cap-dependent translation and reveals a novel regulatory motif. AB - Eukaryotic initiation factor 4E (eIF4E) binding proteins (4E-BPs) regulate the assembly of initiation complexes required for cap-dependent mRNA translation. 4E BP1 undergoes insulin-stimulated phosphorylation, resulting in its release from eIF4E, allowing initiation complex assembly. 4E-BP1 undergoes caspase-dependent cleavage in cells undergoing apoptosis. Here we show that cleavage occurs after Asp24, giving rise to the N-terminally truncated polypeptide Delta4E-BP1, which possesses the eIF4E-binding site and all the known phosphorylation sites. Delta4E BP1 binds to eIF4E and fails to become sufficiently phosphorylated upon insulin stimulation to bring about its release from eIF4E. Therefore, Delta4E-BP1 acts as a potent inhibitor of cap-dependent translation. Using a mutagenesis approach, we identify a novel regulatory motif of four amino acids (RAIP) which lies within the first 24 residues of 4E-BP1 and which is necessary for efficient phosphorylation of 4E-BP1. This motif is conserved among sequences of 4E-BP1 and 4E-BP2 but is absent from 4E-BP3. Insulin increased the phosphorylation of 4E-BP3 but not sufficiently to cause its release from eIF4E. However, a chimeric protein that was generated by replacing the N terminus of 4E-BP3 with the N-terminal sequence of 4E-BP1 (containing this RAIP motif) underwent a higher degree of phosphorylation and was released from eIF4E. This suggests that the N-terminal sequence of 4E-BP1 is required for optimal regulation of 4E-BPs by insulin. PMID- 11865048 TI - Requirements of the RNA polymerase II C-terminal domain for reconstituting pre mRNA 3' cleavage. AB - RNA polymerase II (RNAP II) has previously been shown to be required for the pre mRNA polyadenylation cleavage reaction in vitro. This activity was found to reside solely in the C-terminal domain (CTD) of the enzyme's largest subunit. Using a deletion analysis of glutathione S-transferase-CTD fusion proteins, we searched among the CTD's 52 imperfectly repetitive heptapeptides for the minimal subset that possesses this property. We found that heptads in the vicinity of 30 to 37 contribute modestly more than other sections, but that no specific subsection of the CTD is necessary or sufficient for cleavage. To investigate further the heptad requirements for cleavage, we constructed a series of all consensus CTDs having 13, 26, 39, and 52 YSPTSPS repeats. We found that the nonconsensus CTD heptads are together responsible for only 20% of the wild-type cleavage activity. Analysis of the all-consensus CTD series revealed that the remaining 80% of the CTD-dependent cleavage activity directly correlates with CTD length, with significant activity requiring approximately 26 or more repeats. These results are consistent with a scaffolding role for the RNAP II CTD in the pre-mRNA cleavage reaction. PMID- 11865049 TI - Control of intracellular dynamics of mammalian period proteins by casein kinase I epsilon (CKIepsilon) and CKIdelta in cultured cells. AB - Recent studies have shown that casein kinase I epsilon (CKIepsilon) is an essential regulator of the mammalian circadian clock. However, the detailed mechanisms by which CKIepsilon regulates each component of the circadian negative feedback loop have not been fully defined. We show here that mPer proteins, negative limbs of the autoregulatory loop, are specific substrates for CKIepsilon and CKIdelta. The CKI phosphorylation of mPer1 and mPer3 proteins results in their rapid degradation, which is dependent on the ubiquitin-proteasome pathway. Moreover, CKIepsilon and CKIdelta are able to induce nuclear translocation of mPer3, which requires its nuclear localization signal. The mutation in potential phosphorylation sites on mPer3 decreased the extent of both nuclear translocation and degradation of mPer3 that are stimulated by CKIepsilon. CKIepsilon and CKIdelta affected the inhibitory effect of mPer proteins on the transcriptional activity of BMAL1-CLOCK, but the inhibitory effect of mCry proteins on the activity of BMAL1-CLOCK was unaffected. These results suggest that CKIepsilon and CKIdelta regulate the mammalian circadian autoregulatory loop by controlling both protein turnover and subcellular localization of mPer proteins. PMID- 11865050 TI - The endothelial receptor tyrosine kinase Tie1 activates phosphatidylinositol 3 kinase and Akt to inhibit apoptosis. AB - Tie1 is an orphan receptor tyrosine kinase that is expressed almost exclusively in endothelial cells and that is required for normal embryonic vascular development. Genetic studies suggest that Tie1 promotes endothelial cell survival, but other studies have suggested that the Tie1 kinase has little to no activity, and Tie1-mediated signaling pathways are unknown. To begin to study Tie1 signaling, a recombinant glutathione S-transferase (GST)-Tie1 kinase fusion protein was produced in insect cells and found to be autophosphorylated in vitro. GST-Tie1 but not a kinase-inactive mutant associated with a recombinant p85 SH2 domain protein in vitro, suggesting that Tie1 might signal through phosphatidylinositol (PI) 3-kinase. To study Tie1 signaling in a cellular context, a c-fms-Tie1 chimeric receptor (fTie1) was expressed in NIH 3T3 cells. Ligand stimulation of fTie1 resulted in Tie1 autophosphorylation and downstream activation of PI 3-kinase and Akt. Stimulation of fTie1-expressing cells potently inhibited UV irradiation-induced apoptosis in a PI 3-kinase-dependent manner. Moreover, both Akt phosphorylation and inhibition of apoptosis were abrogated by mutation of tyrosine 1113 to phenylalanine, suggesting that this residue is an important PI 3-kinase binding site. These findings are the first biochemical demonstration of a signal transduction pathway and corresponding cellular function for Tie1, and the antiapoptotic effect of Tie1 is consistent with the results of previous genetic studies. PMID- 11865051 TI - Ral-GTPase influences the regulation of the readily releasable pool of synaptic vesicles. AB - The Ral proteins are members of the Ras superfamily of GTPases. Because they reside in synaptic vesicles, we used transgenic mice expressing a dominant inhibitory form of Ral to investigate the role of Ral in neurosecretion. Using a synaptosomal secretion assay, we found that while K(+)-evoked secretion of glutamate was normal, protein kinase C-mediated enhancement of glutamate secretion was suppressed in the mutant mice. Since protein kinase C effects on secretion have been shown to be due to enhancement of the size of the readily releasable pool of synaptic vesicles docked at the plasma membrane, we directly measured the refilling of this readily releasable pool of synaptic vesicles after Ca(2+)-triggered exocytosis. Refilling of the readily releasable pool was suppressed in synaptosomes from mice expressing dominant inhibitory Ral. Moreover, we found that protein kinase C and calcium-induced phosphorylation of proteins thought to influence synaptic vesicle function, such as MARCKS, synapsin, and SNAP-25, were all reduced in synaptosomes from these transgenic mice. Concomitant with these studies, we searched for new functions of Ral by detecting proteins that specifically bind to it in cells. Consistent with the phenotype of the transgenic mice described above, we found that active but not inactive RalA binds to the Sec6/8 (exocyst) complex, whose yeast counterpart is essential for targeting exocytic vesicles to specific docking sites on the plasma membrane. These findings demonstrate a role for Ral-GTPase signaling in the modulation of the readily releasable pool of synaptic vesicles and suggest the possible involvement of Ral-Sec6/8 (exocyst) binding in modulation of synaptic strength. PMID- 11865052 TI - Promoter scanning for transcription inhibition with DNA-binding polyamides. AB - When targeted to sequences adjacent to a TATA element, pyrrole-imidazole (Py-Im) polyamides inhibit the DNA binding activity of TATA box binding protein (TBP) and basal transcription by RNA polymerase II. In the present study, we scanned the human immunodeficiency virus type 1 promoter for polyamide inhibition of TBP binding and transcription using a series of DNA constructs in which a polyamide binding site was placed at various distances from the TATA box. Polyamide interference with either TBP-DNA or TFIID-TFIIA-DNA contacts both upstream and downstream of the TATA element resulted in inhibition of transcription. Our results define important protein-DNA interactions outside of the TATA element and suggest that transcription inhibition of selected gene promoters can be achieved with polyamides that target unique sequences within these promoters at a distance from the TATA element. Our studies also demonstrate the utility of the Py-Im polyamides for discovery of functionally important protein-DNA contacts involved in transcription. PMID- 11865053 TI - The regulation of hypoxic genes by calcium involves c-Jun/AP-1, which cooperates with hypoxia-inducible factor 1 in response to hypoxia. AB - Hypoxia causes the accumulation of the transcription factor hypoxia-inducible factor 1 (HIF-1), culminating in the expression of hypoxia-inducible genes such as those for vascular endothelial growth factor (VEGF) and NDRG-1/Cap43. Previously, we have demonstrated that intracellular calcium (Ca(2+)) is required for the expression of hypoxia-inducible genes. Here we found that, unlike with hypoxia or hypoxia-mimicking conditions, the elevation of intracellular Ca(2+) neither induced the HIF-1alpha protein nor stimulated HIF-1-dependent transcription. Furthermore, the elevation of intracellular Ca(2+) induced NDRG 1/Cap43 mRNA in HIF-1alpha-deficient cells. It also increased levels of c-Jun protein, causing its phosphorylation. The protein kinase inhibitor K252a abolished c-Jun induction and activator protein 1 (AP-1)-dependent reporter expression caused by Ca(2+) ionophore or hypoxia. K252a also significantly decreased hypoxia-induced VEGF and NDRG-1/Cap43 gene expression in both human and mouse cells. Using a set of deletion VEGF-Luc promoter constructs, we found that both HIF-1 and two AP-1 sites contribute to hypoxia-mediated induction of transcription. In contrast, only AP-1 sites contributed to Ca(2+)-mediated VEGF Luc induction. A dominant-negative AP-1 prevented Ca(2+)-dependent transcription and partially impaired hypoxia-mediated transcription. In addition, dominant negative AP-1 diminished the expression of the NDRG-1/Cap43 gene following hypoxia. We conclude that during hypoxia, an increase in intracellular Ca(2+) activates a HIF-1-independent signaling pathway that involves AP-1-dependent transcription. Cooperation between the HIF-1 and AP-1 pathways allows fine regulation of gene expression during hypoxia. PMID- 11865054 TI - Regulation of nuclear gamma interferon gene expression by interleukin 12 (IL-12) and IL-2 represents a novel form of posttranscriptional control. AB - Posttranscriptional control of gamma interferon (IFN-gamma) gene expression has not been extensively studied and is poorly understood. Our work describes a posttranscriptional mechanism that modulates IFN-gamma mRNA expression in stimulated natural killer (NK) cells through nuclear retention of the IFN-gamma mRNA. This is evidenced by the elevated and sustained nuclear accumulation of both precursor and processed IFN-gamma mRNAs in NK cells stimulated with interleukin-12 (IL-12). The elevated nuclear mRNA accumulation persists long after transcriptional activity has subsided and the rate of cytoplasmic IFN-gamma mRNA accumulation has dropped. The IL-12-induced nuclear retention of the IFN gamma mRNA prevails until a secondary cytokine stimulus is received. The secondary stimulus, which is initiated by IL-2, mediates transcription independent movement of the nuclear IFN-gamma mRNA. Concurrent with the nucleocytoplasmic movement of the IFN-gamma mRNA, we have observed increases in the amount of processed nuclear IFN-gamma mRNA that are greater than that seen for the unprocessed IFN-gamma mRNA. The increase in processed IFN-gamma mRNA appears to be due to increased mRNA stability which then promotes increased nucleocytoplasmic shuttling of the mature IFN-gamma mRNA. These data support a model whereby mobilization of nuclear IFN-gamma mRNA stores allows NK cells to rapidly and robustly respond to secondary cytokine activators in a transcription independent manner, thus shortening the time for overall cellular response to inflammatory signals. PMID- 11865056 TI - Coupling of enhancer and insulator properties identified in two retrotransposons modulates their mutagenic impact on nearby genes. AB - We recently reported a novel transposition system in which two retroelements from Drosophila melanogaster, ZAM and Idefix, are highly mobilized and preferentially insert within intergenic regions. Among the loci where new copies are detected, a hot spot for their insertion was identified at the white locus, where up to three elements occurred within a 3-kb fragment upstream of the transcriptional start site of white. We have used these insertions as molecular entry points to throw light on the mutagenic effect exerted by multiple insertions of retrotransposons within intergenic regions of a genome. Analysis of the molecular mechanisms by which ZAM and Idefix elements interfere with the regulation of the white gene has shown that ZAM bears cis-acting regulatory sequences able to enhance transcription of the white gene in the eyes of the flies. This activation may be counteracted by Idefix, which acts as an insulator able to isolate the white gene from the upstream ZAM enhancer. In addition to revealing a novel insulator sequence with its own specific features, our data clearly illustrate how retroelements can act as epigenetic factors able to interfere with the transcriptional regulation of their host. PMID- 11865055 TI - IAP suppression of apoptosis involves distinct mechanisms: the TAK1/JNK1 signaling cascade and caspase inhibition. AB - The antiapoptotic properties of the inhibitor of apoptosis (IAP) family of proteins have been linked to caspase inhibition. We have previously described an alternative mechanism of XIAP inhibition of apoptosis that depends on the selective activation of JNK1. Here we report that two other members of the IAP family, NAIP and ML-IAP, both activate JNK1. Expression of catalytically inactive JNK1 blocks NAIP and ML-IAP protection against ICE- and TNF-alpha-induced apoptosis, indicating that JNK1 activation is necessary for the antiapoptotic effect of these proteins. The MAP3 kinase, TAK1, appears to be an essential component of this antiapoptotic pathway since IAP-mediated activation of JNK1, as well as protection against TNF-alpha- and ICE-induced apoptosis, is inhibited when catalytically inactive TAK1 is expressed. In addition, XIAP, NAIP, and JNK1 bind to TAK1. Importantly, expression of catalytically inactive TAK1 did not affect XIAP inhibition of caspase activity. These data suggest that XIAP's antiapoptotic activity is achieved by two separate mechanisms: one requiring TAK1 dependent JNK1 activation and the second involving caspase inhibition. PMID- 11865057 TI - A novel and conserved protein-protein interaction domain of mammalian Lin-2/CASK binds and recruits SAP97 to the lateral surface of epithelia. AB - Mammalian Lin-2 (mLin-2)/CASK is a membrane-associated guanylate kinase (MAGUK) and contains multidomain modules that mediate protein-protein interactions important for the establishment and maintenance of neuronal and epithelial cell polarization. The importance of mLin-2/CASK in mammalian development is demonstrated by the fact that mutations in mLin-2/CASK or SAP97, another MAGUK protein, lead to cleft palate in mice. We recently identified a new protein protein interaction domain, called the L27 domain, which is present twice in mLin 2/CASK. In this report, we further define the binding of the L27C domain of mLin 2/CASK to the L27 domain of mLin-7 and identify the binding partner for L27N of mLin-2/CASK. Biochemical analysis reveals that this L27N domain binds to the N terminus of SAP97, a region that was previously reported to be essential for the lateral membrane recruitment of SAP97 in epithelia. Our colocalization studies, using dominant-negative mLin-2/CASK, show that the association with mLin-2/CASK is crucial for lateral localization of SAP97 in MDCK cells. We also report the identification of a novel isoform of Discs Large, a Drosophila melanogaster orthologue of SAP97, which contains a region highly related to the SAP97 N terminus and which binds Camguk, a Drosophila orthologue of mLin-2/CASK. Our data identify evolutionarily conserved protein-protein interaction domains that link mLin-2/CASK to SAP97 and account for their common phenotype when mutated in mice. PMID- 11865058 TI - CKA, a novel multidomain protein, regulates the JUN N-terminal kinase signal transduction pathway in Drosophila. AB - The Drosophila melanogaster JUN N-terminal kinase (DJNK) and DPP (decapentaplegic) signal transduction pathways coordinately regulate epithelial cell sheet movement during the process of dorsal closure in the embryo. By a genetic screen of mutations affecting dorsal closure in Drosophila, we have now identified a multidomain protein, connector of kinase to AP-1 (cka), that functions in the DJNK pathway and controls the localized expression of dpp in the leading-edge cells. We have also investigated how CKA acts. This unique molecule forms a complex with HEP (DJNKK), BSK (DJNK), DJUN, and DFOS. Complex formation activates BSK kinase, which in turn phosphorylates and activates DJUN and DFOS. These data suggest that CKA represents a novel molecule regulating AP-1 activity by organizing a molecular complex of kinases and transcription factors, thus coordinating the spatial-temporal expression of AP-1-regulated genes. PMID- 11865060 TI - Overexpression of c-Myc alters G(1)/S arrest following ionizing radiation. AB - Study of the mechanism(s) of genomic instability induced by the c-myc proto oncogene has the potential to shed new light on its well-known oncogenic activity. However, an underlying mechanism(s) for this phenotype is largely unknown. In the present study, we investigated the effects of c-Myc overexpression on the DNA damage-induced G(1)/S checkpoint, in order to obtain mechanistic insights into how deregulated c-Myc destabilizes the cellular genome. The DNA damage-induced checkpoints are among the primary safeguard mechanisms for genomic stability, and alterations of cell cycle checkpoints are known to be crucial for certain types of genomic instability, such as gene amplification. The effects of c-Myc overexpression were studied in human mammary epithelial cells (HMEC) as one approach to understanding the c-Myc-induced genomic instability in the context of mammary tumorigenesis. Initially, flow-cytometric analyses were used with two c-Myc-overexpressing, nontransformed immortal lines (184A1N4 and MCF10A) to determine whether c-Myc overexpression leads to alteration of cell cycle arrest following ionizing radiation (IR). Inappropriate entry into S phase was then confirmed with a bromodeoxyuridine incorporation assay measuring de novo DNA synthesis following IR. Direct involvement of c-Myc overexpression in alteration of the G(1)/S checkpoint was then confirmed by utilizing the MycER construct, a regulatable c-Myc. A transient excess of c-Myc activity, provided by the activated MycER, was similarly able to induce the inappropriate de novo DNA synthesis following IR. Significantly, the transient expression of full-length c Myc in normal mortal HMECs also facilitated entry into S phase and the inappropriate de novo DNA synthesis following IR. Furthermore, irradiated, c-Myc infected, normal HMECs developed a sub-G(1) population and a >4N population of cells. The c-Myc-induced alteration of the G(1)/S checkpoint was also compared to the effects of expression of MycS (N-terminally truncated c-Myc) and p53DD (a dominant negative p53) in the HMECs. We observed inappropriate hyperphosphorylation of retinoblastoma protein and then the reappearance of cyclin A, following IR, selectively in full-length c-Myc- and p53DD overexpressing MCF10A cells. Based on these results, we propose that c-Myc attenuates a safeguard mechanism for genomic stability; this property may contribute to c-Myc-induced genomic instability and to the potent oncogenic activity of c-Myc. PMID- 11865059 TI - Critical residues within the BTB domain of PLZF and Bcl-6 modulate interaction with corepressors. AB - The PLZF (promyelocytic leukemia zinc finger) transcriptional repressor, when fused to retinoic acid receptor alpha (RARalpha), causes a refractory form of acute promyelocytic leukemia. The highly conserved N-terminal BTB (bric a brac, tramtrack, broad complex)/POZ domain of PLZF plays a critical role in this disease, since it is required for transcriptional repression by the PLZF-RARalpha fusion protein. The crystal structure of the PLZF BTB domain revealed an obligate homodimer with a highly conserved charged pocket formed by apposition of the two monomers. An extensive structure-function analysis showed that the charged pocket motif plays a major role in transcriptional repression by PLZF. We found that mutations of the BTB domain that neutralize key charged pocket residues did not disrupt dimerization, yet abrogated the ability of PLZF to repress transcription and led to the loss of interaction with N-CoR, SMRT, and histone deacetylases (HDACs). We extended these studies to the Bcl-6 protein, which is linked to the pathogenesis of non-Hodgkin's lymphomas. In this case, neutralizing the charged pocket also resulted in loss of repression and corepressor binding. Experiments with purified protein showed that corepressor-BTB interactions were direct. A comparison of the PLZF, Bcl-6, and the FAZF (Fanconi anemia zinc finger)/ROG protein shows that variations in the BTB pocket result in differential affinity for corepressors, which predicts the potency of transcriptional repression. Thus, the BTB pocket represents a molecular structure involved in recruitment of transcriptional repression complexes to target promoters. PMID- 11865061 TI - Hypoxia links ATR and p53 through replication arrest. AB - Previous studies have demonstrated that phosphorylation of human p53 on serine 15 contributes to protein stabilization after DNA damage and that this is mediated by the ATM family of kinases. However, cellular exposure to hypoxia does not induce any detectable level of DNA lesions compared to ionizing radiation, and the oxygen dependency of p53 protein accumulation differs from that of HIF-1, the hypoxia-inducible transcription factor. Here we show that, under severe hypoxic conditions, p53 protein accumulates only in S phase and this accumulation correlates with replication arrest. Inhibition of ATR kinase activity substantially reduces hypoxia-induced phosphorylation of p53 protein on serine 15 as well as p53 protein accumulation. Thus, hypoxia-induced cell growth arrest is tightly linked to an ATR-signaling pathway that is required for p53 modification and accumulation. These studies indicate that the ATR kinase plays an important role during tumor development in responding to hypoxia-induced replication arrest, and hypoxic conditions could select for the loss of key components of ATR dependent checkpoint controls. PMID- 11865062 TI - Precipitous release of methyl-CpG binding protein 2 and histone deacetylase 1 from the methylated human multidrug resistance gene (MDR1) on activation. AB - Overexpression of the human multidrug resistance gene 1 (MDR1) is a negative prognostic factor in leukemia. Despite intense efforts to characterize the gene at the molecular level, little is known about the genetic events that switch on gene expression in P-glycoprotein-negative cells. Recent studies have shown that the transcriptional competence of MDR1 is often closely associated with DNA methylation. Chromatin remodeling and modification targeted by the recognition of methylated DNA provide a dominant mechanism for transcriptional repression. Consistent with this epigenetic model, interference with DNA methyltransferase and histone deacetylase activity alone or in combination can reactivate silent genes. In the present study, we used chromatin immunoprecipitation to monitor the molecular events involved in the activation and repression of MDR1. Inhibitors of DNA methyltransferase (5-azacytidine [5aC]) and histone deacetylase (trichostatin A [TSA]) were used to examine gene transcription, promoter methylation status, and the chromatin determinants associated with the MDR1 promoter. We have established that methyl-CpG binding protein 2 (MeCP2) is involved in methylation dependent silencing of human MDR1 in cells that lack the known transcriptional repressors MBD2 and MBD3. In the repressed state the MDR1 promoter is methylated and assembled into chromatin enriched with MeCP2 and deacetylated histone. TSA induced significant acetylation of histones H3 and H4 but did not activate transcription. 5aC induced DNA demethylation, leading to the release of MeCP2, promoter acetylation, and partial relief of repression. MDR1 expression was significantly increased following combined 5aC and TSA treatments. Inhibition of histone deacetylase is not an overriding mechanism in the reactivation of methylated MDR1. Our results provide us with a clearer understanding of the molecular mechanism necessary for repression of MDR1. PMID- 11865063 TI - The Rab27a/granuphilin complex regulates the exocytosis of insulin-containing dense-core granules. AB - Recently, we identified and characterized a novel protein, granuphilin, whose domain structure is similar to that of the Rab3 effector protein rabphilin3 (J. Wang, T. Takeuchi, H. Yokota, and T. Izumi, J. Biol. Chem. 274:28542-28548, 1999). Screening its possible Rab partner by a yeast two-hybrid system revealed that an amino-terminal zinc-finger domain of granuphilin interacts with Rab27a. Granuphilin preferentially bound to the GTP form of Rab27a. Formation of the Rab27a/granuphilin complex was readily detected in the pancreatic beta cell line MIN6. Moreover, the tissue distributions of Rab27a and granuphilin are remarkably similar: both had significant and specific expression in pancreatic islets and in pituitary tissue, but no expression was noted in the brain. Analyses by immunofluorescence, immunoelectron microscopy, and sucrose density gradient subcellular fractionation showed that Rab27a and granuphilin are localized on the membrane of insulin granules. These findings suggest that granuphilin functions as a Rab27a effector protein in beta cells. Overexpression of wild-type Rab27a and its GTPase-deficient mutant significantly enhanced high K(+)-induced insulin secretion without affecting basal insulin release. Although Rab3a, another exocytotic Rab protein, has some similarities with Rab27a in primary sequence, intracellular distribution, and affinity toward granuphilin, overexpression of Rab3a caused different effects on insulin secretion. These results indicate that Rab27a is involved in the regulated exocytosis of conventional dense-core granules possibly through the interaction with granuphilin, in addition to its recently identified role in lysosome-related organelles. PMID- 11865064 TI - Control of DNA replication and chromosome ploidy by geminin and cyclin A. AB - Alteration of the control of DNA replication and mitosis is considered to be a major cause of genome instability. To investigate the mechanism that controls DNA replication and genome stability, we used the RNA silencing-interference technique (RNAi) to eliminate the Drosophila geminin homologue from Schneider D2 (SD2) cells. Silencing of geminin by RNAi in SD2 cells leads to the cessation of mitosis and asynchronous overreplication of the genome, with cells containing single giant nuclei and partial ploidy between 4N and 8N DNA content. The effect of geminin deficiency is completely suppressed by cosilencing of Double parked (Dup), the Drosophila homologue of Cdt1, a replication factor to which geminin binds. The geminin deficiency-induced phenotype is also partially suppressed by coablation of Chk1/Grapes, indicating the involvement of Chk1/Grapes in the checkpoint control in response to overreplication. We found that the silencing of cyclin A, but not of cyclin B, also promotes the formation of a giant nucleus and overreplication. However, in contrast to the effect of geminin knockout, cyclin A deficiency leads to the complete duplication of the genome from 4N to 8N. We observed that the silencing of geminin causes rapid downregulation of Cdt1/Dup, which may contribute to the observed partial overreplication in geminin-deficient cells. Analysis of cyclin A and geminin double knockout suggests that the effect of cyclin A deficiency is dominant over that of geminin deficiency for cell cycle arrest and overreplication. Together, our studies indicate that both cyclin A and geminin are required for the suppression of overreplication and for genome stability in Drosophila cells. PMID- 11865065 TI - Heparan sulfate proteoglycans are ligands for receptor protein tyrosine phosphatase sigma. AB - RPTPsigma is a cell adhesion molecule-like receptor protein tyrosine phosphatase involved in nervous system development. Its avian orthologue, known as cPTPsigma or CRYPalpha, promotes intraretinal axon growth and controls the morphology of growth cones. The molecular mechanisms underlying the functions of cPTPsigma are still to be determined, since neither its physiological ligand(s) nor its substrates have been described. Nevertheless, a major class of ligand(s) is present in the retinal basal lamina and glial endfeet, the potent native growth substrate for retinal axons. We demonstrate here that cPTPsigma is a heparin binding protein and that its basal lamina ligands include the heparan sulfate proteoglycans (HSPGs) agrin and collagen XVIII. These molecules interact with high affinity with cPTPsigma in vitro, and this binding is totally dependent upon their heparan sulfate chains. Using molecular modelling and site-directed mutagenesis, a binding site for heparin and heparan sulfate was identified in the first immunoglobulin-like domain of cPTPsigma. HSPGs are therefore a novel class of heterotypic ligand for cPTPsigma, suggesting that cPTPsigma signaling in axons and growth cones is directly responsive to matrix-associated cues. PMID- 11865066 TI - Apolipoprotein J/clusterin prevents a progressive glomerulopathy of aging. AB - Apoliprotein J (apoJ)/clusterin has attracted considerable interest based on its inducibility in multiple injury processes and accumulation at sites of remodeling, regression, and degeneration. We therefore sought to investigate apoJ/clusterin's role in kidney aging, as this may reveal the accumulated effects of diminished protection. Aging mice deficient in apoJ/clusterin developed a progressive glomerulopathy characterized by the deposition of immune complexes in the mesangium. Up to 75% of glomeruli in apoJ/clusterin-deficient mice exhibited moderate to severe mesangial lesions by 21 months of age. Wild-type and hemizygous mice exhibited little or no glomerular pathology. In the apoJ/clusterin-deficient mice, immune complexes of immunoglobulin G (IgG), IgM, IgA, and in some cases C1q, C3, and C9 were detectable as early as 4 weeks of age. Electron microscopy revealed the accumulation of electron-dense material in the mesangial matrix and age-dependent formation of intramesangial tubulo fibrillary structures. Even the most extensively damaged glomeruli showed no evidence of inflammation or necrosis. In young apoJ/clusterin-deficient animals, the development of immune complex lesions was accelerated by unilateral nephrectomy-induced hyperfiltration. Injected immune complexes localized to the mesangium of apoJ/clusterin-deficient but not wild-type mice. These results establish a protective role of apoJ/clusterin against chronic glomerular kidney disease and support the hypothesis that apoJ/clusterin modifies immune complex metabolism and disposal. PMID- 11865068 TI - Activating transcription factor 1 and CREB are important for cell survival during early mouse development. AB - Activating transcription factor 1 (ATF1), CREB, and the cyclic AMP (cAMP) response element modulatory protein (CREM), which constitute a subfamily of the basic leucine zipper transcription factors, activate gene expression by binding as homo- or heterodimers to the cAMP response element in regulatory regions of target genes. To investigate the function of ATF1 in vivo, we inactivated the corresponding gene by homologous recombination. In contrast to CREB-deficient mice, which suffer from perinatal lethality, mice lacking ATF1 do not exhibit any discernible phenotypic abnormalities. Since ATF1 and CREB but not CREM are strongly coexpressed during early mouse development, we generated mice deficient for both CREB and ATF1. ATF1(-/-) CREB(-/-) embryos die before implantation due to developmental arrest. ATF1(+/-) CREB(-/-) embryos display a phenotype of embryonic lethality around embryonic day 9.5 due to massive apoptosis. These results indicate that CREB and ATF1 act in concert to mediate signals essential for maintaining cell viability during early embryonic development. PMID- 11865067 TI - Activated Fes protein tyrosine kinase induces terminal macrophage differentiation of myeloid progenitors (U937 cells) and activation of the transcription factor PU.1. AB - The c-fps/fes proto-oncogene encodes a 92-kDa protein tyrosine kinase that is preferentially expressed in myeloid and endothelial cells. Fes is believed to play a role in vascular development and myelopoiesis and in the inflammatory responses of granulocytes and macrophages. To help define the biological role of this kinase and identify its downstream targets, we have developed a gain-of function allele of Fes that has potent biological activity in myeloid cell progenitors. Introduction of constitutively active Fes into bipotential U937 cells induced the appearance of fully differentiated macrophages within 6 to 12 days. The Fes-expressing differentiated cells became adherent, had distinctive macrophage morphology, and exhibited increased expression of myelomonocytic differentiation markers, including CD11b, CD11c, CD18, CD14, and the macrophage colony-stimulating factor receptor. These cells acquired phagocytic properties and exhibited NADPH oxidase and nonspecific esterase activities, confirming that they were functionally active macrophages. Concomitantly, there was downregulation of the granulocytic marker granulocyte colony-stimulating factor receptor, indicating that the biological activity of Fes was coordinated in a lineage-specific manner. A constitutively active Src did not induce macrophage morphology or upregulation of myelomonocytic markers in U937 cells, suggesting that the biological activity we observed was not a general consequence of expression of an activated nonreceptor tyrosine kinase. Analysis of possible downstream targets of Fes revealed that this kinase activated the ets family transcription factor PU.1, which is essential for macrophage development. Our results strongly implicate Fes as a key regulator of terminal macrophage differentiation and identify PU.1 as a transcription factor that may mediate some of its biological activities in myeloid cells. PMID- 11865069 TI - BMAP-28, an antibiotic peptide of innate immunity, induces cell death through opening of the mitochondrial permeability transition pore. AB - BMAP-28, a bovine antimicrobial peptide of the cathelicidin family, induces membrane permeabilization and death in human tumor cell lines and in activated, but not resting, human lymphocytes. In addition, we found that BMAP-28 causes depolarization of the inner mitochondrial membrane in single cells and in isolated mitochondria. The effect of the peptide was synergistic with that of Ca(2+) and inhibited by cyclosporine, suggesting that depolarization depends on opening of the mitochondrial permeability transition pore. The occurrence of a permeability transition was investigated on the basis of mitochondrial permeabilization to calcein and cytochrome c release. We show that BMAP-28 permeabilizes mitochondria to entrapped calcein in a cyclosporine-sensitive manner and that it releases cytochrome c in situ. Our results demonstrate that BMAP-28 is an inducer of the mitochondrial permeability transition pore and that its cytotoxic potential depends on its effects on mitochondrial permeability. PMID- 11865072 TI - Nitric oxide triggers the expression of proinflammatory and protective gene products in mesangial cells and the inflamed glomerulus. PMID- 11865070 TI - Binding of the RING polycomb proteins to specific target genes in complex with the grainyhead-like family of developmental transcription factors. AB - The Polycomb group (PcG) of proteins represses homeotic gene expression through the assembly of multiprotein complexes on key regulatory elements. The mechanisms mediating complex assembly have remained enigmatic since most PcG proteins fail to bind DNA. We now demonstrate that the human PcG protein dinG interacts with CP2, a mammalian member of the grainyhead-like family of transcription factors, in vitro and in vivo. The functional consequence of this interaction is repression of CP2-dependent transcription. The CP2-dinG interaction is conserved in evolution with the Drosophila factor grainyhead binding to dring, the fly homologue of dinG. Electrophoretic mobility shift assays demonstrate that the grh dring complex forms on regulatory elements of genes whose expression is repressed by grh but not on elements where grh plays an activator role. These observations reveal a novel mechanism by which PcG proteins may be anchored to specific regulatory elements in developmental genes. PMID- 11865074 TI - Managing big issues on lean evidence: treating obesity hypertension. PMID- 11865073 TI - Statins and angiotensin II-induced vascular injury. PMID- 11865071 TI - Diverse effects of mutations in exon II of the von Hippel-Lindau (VHL) tumor suppressor gene on the interaction of pVHL with the cytosolic chaperonin and pVHL dependent ubiquitin ligase activity. AB - We examined the biogenesis of the von Hippel-Lindau (VHL) tumor suppressor protein (pVHL) in vitro and in vivo. pVHL formed a complex with the cytosolic chaperonin containing TCP-1 (CCT or TRiC) en route to assembly with elongin B/C and the subsequent formation of the VCB-Cul2 ubiquitin ligase. Blocking the interaction of pVHL with elongin B/C resulted in accumulation of pVHL within the CCT complex. pVHL present in purified VHL-CCT complexes, when added to rabbit reticulocyte lysate, proceeded to form VCB and VCB-Cul2. Thus, CCT likely functions, at least in part, by retaining VHL chains pending the availability of elongin B/C for final folding and/or assembly. Tumor-associated mutations within exon II of the VHL syndrome had diverse effects upon the stability and/or function of pVHL-containing complexes. First, a pVHL mutant lacking the entire region encoded by exon II did not bind to CCT and yet could still assemble into complexes with elongin B/C and elongin B/C-Cul2. Second, a number of tumor derived missense mutations in exon II did not decrease CCT binding, and most had no detectable effect upon VCB-Cul2 assembly. Many exon II mutants, however, were found to be defective in the binding to and subsequent ubiquitination of hypoxia inducible factor 1alpha (HIF-1alpha), a substrate of the VCB-Cul2 ubiquitin ligase. We conclude that the selection pressure to mutate VHL exon II during tumorigenesis does not relate to loss of CCT binding but may reflect quantitative or qualitative defects in HIF binding and/or in pVHL-dependent ubiquitin ligase activity. PMID- 11865075 TI - Erythropoietin, tumours and the von Hippel-Lindau gene: towards identification of mechanisms and dysfunction of oxygen sensing. PMID- 11865076 TI - Epoetin treatment: what are the arguments to expect a beneficial effect on renal disease progression? PMID- 11865077 TI - The inflammatory component in progressive renal disease--are interventions possible? PMID- 11865078 TI - Sexual hormone abnormalities in male patients with renal failure. PMID- 11865080 TI - The anastomosis of the arteriovenous fistula--common errors and their avoidance. PMID- 11865079 TI - Late referral: loss of chance for the patient, loss of money for society. PMID- 11865081 TI - Guidelines by an ad hoc European committee on adequacy of the paediatric peritoneal dialysis prescription. PMID- 11865082 TI - Influence of high glucose concentrations on the expression of glycosaminoglycans and N-deacetylase/N-sulphotransferase mRNA in cultured skin fibroblasts from diabetic patients with or without nephropathy. AB - BACKGROUND: The Steno hypothesis postulates that a genetic defect in the regulation of the production of heparan sulphate by renal and non-renal cells determines susceptibility for the development of proteinuria and macro-angiopathy in patients with diabetic nephropathy (DN). METHODS: To test this hypothesis, skin fibroblasts isolated from type II diabetic patients with overt DN, micro albuminuria, or without DN and from non-diabetic patients (n=8 for each group) were cultured in the presence of 5 or 25 mM D-glucose or in 25 mM L-glucose, and tested for the expression of N-deacetylase/N-sulphotransferase (NDST) 1 and 2 by semi-quantitative RT--PCR. Proteoglycan production was measured by means of metabolic labelling. RESULTS: In each group of patients, 25 mM D-glucose significantly reduced the incorporation of [3H]glucosamine (P<0.01), but not [35S]sulphate. The quantity of NDST 1 mRNA expression did not differ between the four groups. In the non-diabetic group only, 25 mM D-glucose significantly increased NDST 1 mRNA expression (P<0.01). In contrast, NDST 2 mRNA expression was reduced by 25 mM D-glucose in all groups (P<0.01). In the diabetic patients, NDST 2 mRNA was significantly reduced compared with the non-diabetic patients. No differences were found between patients with or without nephropathy. In mesangial cells (MC), NDST expression was not influenced by glucose. CONCLUSIONS: Since NDST 1 and 2 are not differentially expressed in patients with or without nephropathy and, in MC, the mRNA expression hereof is not influenced by glucose as in skin fibroblasts, our data do not support the Steno hypothesis. PMID- 11865083 TI - Mechanism of vascular smooth muscle cells activation by hydrogen peroxide: role of phospholipase C gamma. AB - BACKGROUND: Hydrogen peroxide (H2O2) formation is a critical factor in processes involving ischaemia/ reperfusion. However, the precise mechanism by which reactive oxygen species (ROS) induce vascular damage are insufficiently known. Specifically, activation of phospholipase C gamma (PLCgamma) is a probable candidate pathway involved in vascular smooth muscle cells (VSMC) activation by H2O2. METHODS: The activation of human venous VSMC was measured as cytosolic free calcium mobilization, shape change and protein phosphorylation, focusing on the role of tyrosine phosphorylation-activated PLCgamma. RESULTS: The exposure of VSMC to exogenous H2O2 caused a rapid increase in cytosolic free calcium concentration ([Ca2+]i), and induced a significant VSMC shape change. Both effects were dependent on a tyrosine kinase-mediated mechanism, as determined by the blockade of short-term treatment of VSMC with the protein tyrosine kinase inhibitor, genistein. Giving further support to the putative role of phospholipase C (PLC)-dependent pathways, the [Ca2+]i and VSMC shape change response were equally inhibited by the specific PLC blocker, 1-(6-((17-beta methoxyestra-1,3,5(10)trien-17-yl)amino)hexyl)-1H-pyrrole-2,5-dione (U73122). In addition, U73122 had a protective effect against the deleterious action (24 h) of H2O2 on non-confluent VSMC. As a further clarification of the specific pathway involved, the exposure to H2O2 significantly stimulated the tyrosine phosphorylation of PLCgamma with a concentration- and time-profile similar to that of [Ca(2+)](i) mobilization. CONCLUSIONS: The present study reveals that H(2)O(2) activates PLCgamma on VSMC through tyrosine phosphorylation and that this activation has a major role in rapid [Ca(2+)](i) mobilization, shape changing actions and damage by H(2)O(2) in this type of cells. These findings have direct implications for understanding the mechanisms of the vascular actions of H(2)O(2) and may help to design pharmacologically protective strategies. PMID- 11865084 TI - Administration of FR167653, a new anti-inflammatory compound, prevents renal ischaemia/reperfusion injury in mice. AB - BACKGROUND: Various types of chemokines/cytokines play important roles in ischaemia/reperfusion injury in kidneys. However, the roles of p38 mitogen activated protein kinase (MAPK) in the inflammatory processes of renal ischaemia/reperfusion injury remain to be investigated. We explored the effect of FR167653, a specific inhibitor of p38 MAPK, on renal ischaemia/reperfusion injury in mice. METHODS: The renal artery and vein of the left kidney were occluded with a vascular clamp for 60 min. FR167653 was injected 2 h before or 24 h after renal vessel clamp. Renal tissues were removed for pathological examination 4, 24 or 48 h after reperfusion. RESULTS: We observed a large number of infiltrated cells and marked acute tubular necrosis in outer medulla after renal ischaemia/reperfusion injury in mice. FR167653 significantly decreased cell infiltration into outer medulla, and the extent of acute tubular necrosis 24 and 48 h after reperfusion. FR167653 markedly decreased the transcription of interleukin-1beta, tumour necrosis factor-alpha, monocyte chemoattractant protein-1 and regulated upon activation, normal T cell expression and secreted in diseased kidneys. Moreover, FR167653 decreased the number of phosphorylated p38 MAPK-positive cells 4 h after reperfusion. CONCLUSION: These results suggest that FR167653 markedly ameliorated renal ischaemia/reperfusion injury, possibly by inhibiting cytokine/chemokine expression and consequent phosphorylation of p38 MAPK in renal tissue. PMID- 11865085 TI - Progression rate of Chinese herb nephropathy: impact of Aristolochia fangchi ingested dose. AB - BACKGROUND: Renal failure after ingestion of Chinese herbs between 1990 and 1992 was related to the replacement of Stephania tetrandra by Aristolochia fangchi (ST AF), containing nephrotoxic and carcinogenic aristolochic acids. However, the relationship between ST-AF and renal failure is still a matter of debate. We therefore tested the impact of the ST-AF ingested dose on the progression of renal function deterioration. METHODS: Analysis of medical charts and prescriptions between 1990 and 1992 was carried out to determine the presence of risk factors for kidney failure and the cumulative dose of pill components. Individual progression rate of renal impairment was studied by the time-course of the inverse of blood creatinine level (1/P(creat)). RESULTS: Patients were divided into an end-stage renal disease (ESRD) group (n=44) and a chronic renal failure (CRF) group (n=27) according to their P(creat) at the time of this study. The mean number of risk factors (+/-SD) was equally distributed within both groups (1.50+/-0.18 vs 1.59+/-0.17, P=0.74). Patients from the ESRD group ingested significantly higher cumulative doses of ST--AF (192+/-13.1 g vs 138+/- 16.3 g), Magnolia officinalis, (80.1+/-6.3 g vs 59.8+/-11.7 g), diethylpropion (14.7+/-1.4 g vs 10.0+/-1.4 g) and fenfluramine (14.1+/-1.6 g vs 8.7+/-1.3 g). In the ESRD group, some patients who had received steroids had a slower progression to ESRD than the others. In multiple regression analysis, ST-AF emerged as the only significant drug predicting the slope of the progression of renal failure. Moreover, hypothesizing a linear dose-response relationship, the risk of developing ESRD linearly increased with ST-AF doses. CONCLUSIONS: The relationship between the cumulative ST-AF dose and the renal failure progression rate confirms that regular ingestion of Aristolochia sp. extracts is causally involved in the onset of chronic interstitial nephropathy leading to ESRD. PMID- 11865086 TI - Free amino acids in plasma, red blood cells, polymorphonuclear leukocytes, and muscle in normal and uraemic children. AB - BACKGROUND: The aims of this study were to investigate free amino acid (AA) concentrations in plasma, red blood cells (RBC), polymorphonuclear granulocytes (PMN), and muscle, sampled at the same time, in normal and uraemic children. METHODS: Twelve apparently well-nourished chronically uraemic children (five females) aged a mean of 9.4+/-4.8 (range 1.7--17.7) years and 13 age-matched normal children were studied. Venous blood and muscle samples for AA analyses were taken simultaneously after an overnight fast. RESULTS: The intracellular AA patterns in the three cellular compartments were qualitatively similar, but the absolute intracellular concentrations were higher in muscle than in PMN, which had higher values than in RBC. The AA patterns in plasma, RBC, PMN, and muscle in the uraemic children have many similarities; typical features being low branched chain AA (BCAA), tyrosine, and serine concentrations and variably high concentrations of some non-essential AA. Among the individual AA, there were only few correlations between their concentrations in the three cell compartments. CONCLUSIONS: The lack of correlation between the concentrations in RBC, PMN, and muscle for most of the AA indicates that there is no close association in the same subject between individual free AA concentrations in various types of cells, presumably because of differences in metabolism and function. Consequently, one should be cautious in assuming that AA concentrations, determined in RBC or PMN, reflect the concentrations in muscle cells. Therefore, these preliminary observations do not support the hypothesis that RBC and PMN AA analysis can be considered as a suitable alternative to muscle AA determination. PMID- 11865087 TI - Convective and diffusive losses of vitamin C during haemodiafiltration session: a contributive factor to oxidative stress in haemodialysis patients. AB - BACKGROUND: Enhanced oxidative stress in haemodialysis (HD) patients may be considered as a risk factor for accelerated atherosclerosis. Reduced antioxidant defences include impairment in enzyme activities and decreased plasma levels of hydrophilic vitamin C (vit C), and cellular levels of lipophilic vitamin E (vit E). METHODS: We investigated plasma levels of vit C in 19 patients undergoing regular haemodiafiltration (HDF) (mean age 62+/-7 years) and in 1846 healthy elderly subjects (HS) (mean age 69+/-5 years). The contribution of convection and diffusion was determined using paired filtration dialysis (PFD), a modified HDF technique which physically separates convective from diffusive fluxes. Blood samples were collected before and after the HDF session; in addition at 60 min of HDF, samples were drawn from arterial lines (AL) and venous lines (VL), dialysate (D) and ultrafiltrate (UF). Blood levels of total vit C were determined using an HPLC fluorescence method. Markers of oxidative stress were also assessed in both populations as follows: levels of malondialdehyde (MDA) were determined by fluorometric assay, measurements of advanced oxidation protein products (AOPP) and glutathione peroxidase (GSH-Px) activity were performed by spectrophotometric assay, and plasma vit E content was obtained by an HPLC procedure. RESULTS: A significant reduction in plasma vit C level was observed in HDF patients when compared with HS (1.6+/-1.4 microg/ml in HDF vs 6.6+/-3.7 microg/ml in HS; P<0.01). The HDF session was associated with a dramatic reduction in vit C levels (1.87+/-1.57 microg/ml before HDF and 0.98+/-0.68 microg/ml after HDF); at 60 min of HDF, concentrations were as follows: AL=1.35+/-1.27 microg/ml; VL=0.37+/-0.31 microg/ml, D=0.40+/-0.34 microg/ml, UF=1.24+/-1.18 microg/ml; corresponding to a diffusive flux of 271 microg/min and a convective flux of 126 microg/min. Total loss of vit C could be assessed at 66 mg/session (8--230 mg/session). According to this loss of vit C, presence of an oxidative stress was demonstrated in HD population as shown by a significant increase in MDA (1.66+/-0.27 microM in HD vs 0.89+/-0.25 microM in HS; P<0.01) and AOPP (77.5+/-29.3 microM in HD vs 23.5+/ 13.2 microM in HS; P<0.01) levels, and a decrease in GSH-Px activity (259.2+/ 106.3 U/l in HD vs 661.2+/-92.2 U/l in HS; P<0.01). No change in plasma vit E between both populations (30.7+/-9.1 microM in HD vs 35.3+/-7.34 microM in HS) was observed. CONCLUSIONS: These results suggest that HDF with highly permeable membranes is associated with a significant loss of vit C. Diffusive transport is responsible for two-thirds whereas convective phenomenon accounts for only one third of this loss. PMID- 11865088 TI - The effect of LPS, uraemia, and haemodialysis membrane exposure on CD14 expression in mononuclear cells and its relation to apoptosis. AB - BACKGROUND: Both uraemia and bioincompatible haemodialysis membranes induce mononuclear cell apoptosis. Recent reports demonstrate that spontaneous apoptosis in normal monocytes is associated with the down-regulation of CD14 molecules, whereas LPS which prevents the down-regulation of CD14 favours monocyte survival. The aim of the present study was to evaluate a possible association between mononuclear cell apoptosis and low expression of CD14 molecules. This study also investigated whether LPS affects mononuclear cell CD14 expression and the apoptosis induced by uraemia and exposure to Cuprophan (CU) membrane. METHODS: The study was performed in vitro examining the effects of CU membrane and LPS on mononuclear cells from normal subjects and from end-stage renal failure patients. Cells were analysed by flow cytometry with fluorescent monoclonal antibodies to determine CD14 expression and with Annexin-V labelling to determine apoptosis. RESULTS: In mononuclear cells from uraemic patients cultured for 48 h, there was a subset of cells with low CD14 expression; this subset of cells was not observed in normal monocytes cultured for the same period of time. Cells with low CD14 expression were also observed when normal or uraemic mononuclear cells were cultured in the presence of CU membrane. Simultaneous measurement of apoptosis and CD14 expression revealed that cells with low CD14 expression underwent apoptosis. The addition of LPS to the medium markedly reduced the number of mononuclear cells with low CD14 expression and also reduced the rate of apoptosis in these cells. CONCLUSION: Our data suggest that mononuclear cell apoptosis induced by uraemia and the CU membrane is associated with low CD14 expression. Furthermore, LPS prevented the decrease in CD14 and reduced the rate of apoptosis. PMID- 11865089 TI - Circulating levels of ICAM-1, VCAM-1, and MCP-1 are increased in haemodialysis patients: association with inflammation, dyslipidaemia, and vascular events. AB - BACKGROUND: Increased levels of circulating adhesion molecules and chemokines have been reported in haemodialysis (HD) patients but the influence of the HD membranes on their secretion, as well as their pathophysiological implications, remains largely unknown. METHODS: Circulating levels of intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), and monocyte chemoattractant protein-1 (MCP-1) were measured by immunosorbent assay (ELISA) in 81 HD patients (45 male, mean age 57+/-13 years) and 35 normal subjects. All patients had been stabilized on renal replacement therapy for >3 months and were free of active infection. Thirty-three patients (40.7%) were routinely dialysed with modified cellulose membranes and 48 patients (59.3%) were dialysed with polysulfone membranes. Blood samples were taken directly from the arteriovenous fistula immediately before and at the end of a routine HD session. RESULTS: Pre dialysis levels were significantly elevated in HD patients compared with controls (ICAM-1 515+/-177 vs 238+/-664 ng/ml, P<0.0001; VCAM-1 2107+/-648 vs 1012+/-115 ng/ml, P<0.0001; MCP-1 427+/-148 vs 125+/-42 pg/ml, P<0.0001). The HD session resulted in a significant increase in the levels of all three molecules measured (515+/-177 vs 679+/-187 ng/ml, P<0.0001; 2107+/-648 vs 2662+/-800 ng/ml, P<0.0001; 427+/-148 vs 567+/-153 pg/ml, P<0.0001, respectively). There was no difference in pre- or post-dialysis levels of the above molecules between patients routinely dialysed with either modified cellulose or polysulfone membranes. MCP-1 levels had a positive correlation with ICAM-1 levels (r=0.41, P<0.0005). VCAM-1 levels had a negative correlation with HDL levels (r=-0.30, P<0.01) and were significantly elevated in patients with HDL <35 mg/dl compared with patients with HDL > or = 35 mg/dl (2300+/-606 vs 1890+/-633 ng/ml, P<0.005). Log-transformed exact C-reactive protein (CRP) values were significantly correlated with ICAM-1 and VCAM-1 levels (r=0.41, P<0.005 and r=0.43, P<0.005, respectively). In addition, compared with patients with normal CRP values, patients with elevated CRP had significantly increased levels of ICAM-1 (466+/ 166 vs 580+/-172 ng/ml, P<0.005). Patients with cardiovascular, cerebrovascular, or peripheral vascular diseases had significantly increased serum CRP and ICAM-1 levels compared with patients with no evidence of vascular disease (19.2+/-12.9 vs 7.9+/-11.8 mg/l, P<0.001 and 608+/-189 vs 474+/-155 ng/ml, P<0.005 respectively). CONCLUSIONS: Serum levels of ICAM-1, VCAM-1, and MCP-1 are increased in HD patients and probably result from either inadequate clearance or enhanced synthesis and release. HD session resulted in a significant increase of the above molecule levels but the exact mechanism(s) responsible for these alterations are yet to be fully elucidated. Increased levels of adhesion molecules are associated with inflammation, dyslipidaemia, and cardiovascular events. However, the potential link between these processes and its clinical significance warrants further investigation. PMID- 11865090 TI - Muscle mass index in haemodialysis patients: a comparison of indices obtained by routine clinical examinations. AB - BACKGROUND: Measurement of muscle mass is useful for evaluating protein nutritional status. Various methods for estimating muscle mass in haemodialysis patients have recently been developed. METHODS: The validity of the estimate of creatinine production calculated with the creatinine kinetic model (CKM) was examined in 46 haemodialysis patients by comparing it with the actual creatinine production, this being determined from the sum of creatinine appearing in the dialysate and the estimated metabolic degradation. The correlation of various other muscle mass indices with creatinine production was also investigated in these patients. RESULTS: The estimate of creatinine production using CKM was significantly correlated with creatinine production calculated from the spent dialysate plus an estimate for the extra-renal creatinine degradation (r=0.90, P<0.001). A Bland-Altman analysis revealed that the mean prediction error for the estimate of creatinine production by CKM was +0.10 g/day and the limits of agreement were +0.34 to -0.14 g/day. The cross-sectional area of the thigh muscle measured by computed tomography (CT) was also significantly correlated with creatinine production (r=-0.86, P<0.01). In contrast, the correlations of 3 methylhistidine production measured in the spent dialysate, the mid-upper arm muscle circumference and the skeletal muscle mass estimated by an anthropometric prediction model with creatinine production were lower (r<0.82). CONCLUSION: Creatinine production calculated using CKM and CT measurement of thigh muscle area are valid methods for estimating muscle mass during routine clinical examinations of haemodialysis patients. PMID- 11865091 TI - Fluoride and strontium accumulation in bone does not correlate with osteoid tissue in dialysis patients. AB - BACKGROUND: Osteomalacia is now a rare disease in dialysis patients in developed countries since the withdrawal of aluminium overload. The involvement of fluoride and strontium in the pathogenesis of the disease has been suggested. The aim of this study was to investigate a possible association between osteomalacia in dialysis patients and the fluoride or strontium contents of bone. METHODS: Of 271 bone biopsies from chronic haemodialysis patients referred to our centre, we studied the nine biopsies from patients with osteomalacia. They were compared with 23 biopsies from patients with hyperparathyroidism and 24 biopsies from patients with adynamic bone disease. Histomorphometric static and dynamic indices were measured. Bone fluoride and strontium contents were measured in biopsies from haemodialysis patients, and were compared with those of control patients. RESULTS: In the nine patients with osteomalacia, we found an absence of double labelled surfaces and increased osteoid thickness. Mild aluminium overload was observed in two of the nine patients. The bone strontium content of the entire dialysis population studied was not significantly different from control values (0.023+/-0.001 vs 0.019+/-0.002% mol/mol, P=0.15). However, bone strontium level was slightly but significantly increased in patients with osteomalacia (0.030+/ 0.005%), compared with both controls (0.019+/-0.002%, P<0.05) and the other bone diseases (0.021+/-0.002%, P<0.05). Bone fluoride content was significantly higher in the entire dialysis population than in the controls (0.33+/-0.04 vs 0.13+/ 0.018% (g/g ash weight), P=0.04). It was increased in osteomalacic patients compared with controls and with patients having hyperparathyroidism or adynamic bone disease. There was no correlation between formation indices (OV/BV, OS/BS, Ob.S/BS) and bone fluoride or strontium content. CONCLUSIONS: We found a prevalence of osteomalacia of 3.3% in our biopsy series for chronic dialysis patients. However, although bone strontium and fluoride contents were slightly increased, no causal relationship with these individual metals and osteomalacia could be firmly established in this small number of patients. The hypothesis of strontium- or fluoride-induced osteomalacia in renal patients merits further investigation. PMID- 11865092 TI - Hyperhomocysteinaemia, folate and vitamin B12 in unsupplemented haemodialysis patients: effect of oral therapy with folic acid and vitamin B12. AB - BACKGROUND: Hyperhomocysteinaemia, a risk factor for atherosclerosis, is common in dialysis patients and particularly in those homozygous for a common polymorphism in the 5,10-methylenetetrahydrofolate reductase (MTHFR) gene (C677T transition). B-complex vitamin supplements have been shown to lower plasma total homocysteine (tHcy) concentrations, but the respective effectiveness of folate and oral vitamin B12 is not yet known. Our objectives were: (i) to determine the status of folate and vitamin B12 in a cohort of unsupplemented dialysis patients (ii) to assess the homocysteine-lowering effect of a folate supplement and then of a folate supplement with added vitamin B12. The responses were analysed for the C677T genotypes of MTHFR. METHODS: Plasma tHcy, folate and vitamin B12 were measured in 51 haemodialysis patients genotyped for the C677T MTHFR mutation (homozygotes, TT; heterozygotes, CT; without mutation, CC). All patients were then given daily supplements of 15 mg of folic acid for 2 months. They were given daily supplements of 1 mg of vitamin B12 in addition to the folate supplements for a further 2 months. Plasma tHcy, folate and vitamin B12 were monitored after each intervention. RESULTS: At baseline folate and vitamin B12 deficiencies were found in 10% and 6% of the patients. Initial plasma tHcy concentrations were high in all patients (mean 38.1+/-15 micromol/l). CC patients tended to have a lower tHcy concentration than pooled CT and TT patients. After 2 months of folate therapy, tHcy concentration decreased significantly to 20.2+/-7 micromol/l (P<0.001) and no significant differences were observed between the different genotype subgroups (19.4+/-6 for CC, 21.3+/-8 for CT, 18.5+/-4 for TT). A significant positive relationship was found between the reduction of tHcy and its initial value (rho=0.615, P<0.0001). The impact of the added vitamin B12 was negligible since tHcy concentrations did not change for the patients as a whole (19.8+/-7 micromol/l, NS) or in any subgroup (19.1+/-5 for CC, 20.3+/-9 for CT and 20+/-7 micromol/l for TT). CONCLUSIONS: (i) Folate and vitamin B12 deficiencies were observed in 10% and 6% respectively of our unsupplemented dialysis patients. (ii) After folate therapy, tHcy levels decreased significantly in all patients and were identical between the three C677T MTHFR genotype subgroups. (iii) Vitamin B12 supplements are useful in folate treated patients to prevent cobalamin deficiency and its neurological consequences but they did not lower tHcy plasma levels for the patients as a group or for any of the MTHFR subgroups. PMID- 11865093 TI - Detection of microemboli in the subclavian vein of patients undergoing haemodialysis and haemodiafiltration using pulsed Doppler ultrasound. AB - BACKGROUND: The pathophysiology leading to pulmonary side effects during haemodialysis and haemodiafiltration is not yet fully understood. Chronic microembolization, which can be demonstrated by pulsed Doppler ultrasound, may be one cause. METHODS: The study cohort consisted of 24 long-term dialysis patients undergoing haemodialysis (n=21) and online-haemodiafiltration (n=3), respectively. The subclavian vein downstream to the venous access was investigated during different phases of the procedure using a 2-MHz pulsed ultrasound device. RESULTS: In all periods investigated (connection, dialysis, disconnection), numerous microembolic signals (MES) were found in the subclavian vein. The numbers of MES detected during haemodiafiltration (314-709 MES per 10 min) were higher than during haemodialysis (0-81 MES per 10 min). CONCLUSIONS: The composition (gaseous or solid) and origin (pump, tubing system or shunt) of the microemboli detected remains unclear. Chronic microembolization may be one cause of pulmonary complications of haemodialysis and haemodiafiltration. The detection method described in this article will help us to better understand this process and to determine what role microemboli might play in pulmonary and central nervous system disorders. It may also help to optimize the devices and techniques used. PMID- 11865094 TI - Percutaneous thrombolysis of thrombosed haemodialysis access grafts: comparison of three mechanical devices. AB - BACKGROUND: Percutaneous thrombolysis has become an accepted treatment of thrombosed haemodialysis grafts. Several devices have been developed for mechanical thrombolysis, which macerate the clot using different mechanisms such as aspiration and fragmentation. The aim of our study was to compare the efficacy of three devices for mechanical thrombolysis in removing the thrombus from thrombosed haemodialysis access grafts and to determine the initial technical and clinical success, complication rates of each device, and graft patency after the procedure. METHODS: Thrombolysis (i.e. clot removal followed by percutaneous transluminal angioplasty (PTA)) was performed in 68 thrombosed haemodialysis grafts using the Cragg brush catheter combined with urokinase in 13, the Hydrolyser in 18 and the Arrow-Trerotola Percutaneous Thrombolytic Device (PTD) in 37. Clot removal scores (CRS, the ability to thoroughly remove clot from the access), initial technical success, clinical success, patency at 30, 60, and 90 days, and complication rates were evaluated. RESULTS: CRS for the Cragg brush, Hydrolyser and PTD were good in 92, 44, and 95% of cases, respectively. Initial technical (85, 83, and 95%) and clinical success (62, 67, and 86%), mean patency rates at 30 (73, 60, and 55%), 60 (61, 53, and 49%), and 90 (49, 40, and 43%) days, stenosis after PTA (33, 46, and 21%) and complication rates (8, 6, and 0%) were not different for the three devices. Success rates and graft patency depended on the effect of PTA, irrespective of the device used. CONCLUSIONS: The rotational devices removed clots more effectively than the Hydrolyser, with the PTD having the advantage of not requiring urokinase. However, the result of PTA in the treatment of underlying stenoses was the only predictive value for graft patency. PMID- 11865095 TI - Attitudes of British Isles nephrologists towards dialysis modality selection: a questionnaire study. AB - BACKGROUND: Dialysis demographics are changing around the world. Within the UK a striking decrease in the overall use of peritoneal dialysis (PD) has been noted. We set out to determine the opinions and attitudes of British Isles nephrologists about dialysis modality decisions and optimal dialysis system design. METHODS: A survey questionnaire was mailed to a random selection of members of the Renal Association of Great Britain and Ireland. RESULTS: A 63% response rate was achieved. Decisions about dialysis modality were based mostly on patient preference (mean score 4.4 on a scale of 1-5), quality of life data (mean score 3.8), and morbidity and mortality data (mean scores for both 3.6). The least important factors when choosing the modality of dialysis care were the treatment costs to either the patient or the health care system. Respondents felt that both PD and hospital-based haemodialysis (HD) were over-utilized in today's practice. They suggested that an 'ideal dialysis system' (based on patient survival, wellness, and quality of life) should have 27% of patients dialysed using hospital-based HD, 24% in a satellite unit, 11% dialysed using home HD, and 38% on some form of PD (19, 16, and 3% for CAPD, automated PD and intermittent PD, respectively). Few differences were identified between an ideal system which optimized patient survival, wellness, and quality of life, compared with one which optimized cost-effectiveness. CONCLUSION: This survey suggests that most nephrologists in the British Isles feel that hospital-based HD and CAPD are being currently overused, and that future dialysis planning should include a higher proportion of patients on satellite dialysis, home HD, and automated PD to optimize both dialysis cost-effectiveness and patient outcomes. PMID- 11865097 TI - Soluble Fas is a marker of peripheral arterial occlusive disease in haemodialysis patients. AB - BACKGROUND: Peripheral arterial occlusive disease (PAOD) including lower extremity and cerebrovascular atherosclerosis is a leading cause of morbidity in haemodialysis patients. Recent evidence suggests that the expression of Fas, a molecule implicated in the initiation of apoptosis in various cell types, is increased at sites of atherosclerotic plaques. However, the significance of plasma levels of the soluble form of Fas (sFas) as a marker of peripheral arterial disease has yet to be defined. METHODS: The present report is based on a cross-sectional analysis of baseline data from an ongoing prospective study designed to evaluate the role of sFas as marker of PAOD in end-stage renal disease (ESRD). We evaluated the association between sFas levels and evidence of PAOD in a cohort of 107 chronic haemodialysis patients. RESULTS: Compared with subjects without evidence of disease (n=56), subjects with PAOD (n=51) had significantly higher plasma levels of sFas (30.0+/-8.9 vs 26.4+/-9.5 ng/ml; P=0.04). Using multiple regression, sFas was found to be associated with PAOD independently of classical risk factors for atherosclerosis (hypercholesterolaemia, diabetes, hypertension, and smoking), markers of inflammation (e.g. C-reactive protein, intercellular cell adhesion molecule type 1), and other risk factors (e.g. age, gender). An increase of one quintile in the plasma concentration of sFas was associated with an odds ratio of PAOD of 1.69 (95% CI: 1.09--2.63, P=0.01). In addition, models that incorporated sFas were significantly better at predicting PAOD than models limited to classical risk factors for atherosclerosis, alone or in combination with CRP levels (P=0.01). CONCLUSIONS: Increased plasma levels of sFas are associated with established PAOD. These results suggest that sFas may represent a novel and independent marker of atherosclerosis. PMID- 11865096 TI - Expression of nephrin in acquired human glomerular disease. AB - BACKGROUND: Nephrin is a recently identified protein, which is synthesized in the podocytes and localized in the slit diaphragm area. Nephrin is a cell adhesion molecule of the immunoglobulin superfamily, and presumably is a part of the zipper-like structure of the slit membrane. As the mutation of the gene coding nephrin induces congenital nephrotic syndrome of Finnish type, which is a prototype of nephrotic syndrome, it has been suggested that nephrin also plays a role in acquired proteinuric kidney disease. METHODS: To address the above issue, the expression of nephrin in acquired human glomerular disease was studied by immunoelectron microscopy employing a polyclonal antibody against nephrin. Four normal human kidneys from nephrectomy specimens and eight kidney biopsy specimens from glomerular disease patients (one minimal change disease, one membranous glomerulonephritis (GN), one membranoproliferative GN, four IgA nephropathy, and one lupus nephritis) were studied. Proteinuria of the patients ranged from 448 to 11725 mg/day. Effacement of the foot processes was observed in all patients. RESULTS: The study demonstrated that the number and distribution of gold particles in the glomerular region, where the podocyte foot process was well preserved, were similar to that found in normal kidneys; however, gold particles were almost always absent in regions where the foot processes were effaced. The number of gold particles per foot process interspace was not different between normal controls and GN patients; however, the number of gold particles per defined length (1000 nm) of the glomerular basement membrane underlying the foot processes was significantly reduced in GN patients. CONCLUSION: Using immunoelectron microscopy, we observed that the expression of nephrin in GN was lower in regions where the foot processes were effaced, and comparable with that of normal controls where the foot process interspaces were preserved. The significance of our observation in the context of proteinuria in acquired GN needs further clarification. PMID- 11865098 TI - An unusual evolution of the systemic capillary leak syndrome. PMID- 11865099 TI - Mycoplasma hominis infection in renal transplantation. PMID- 11865100 TI - A fatal case of bowel and cardiac involvement in Henoch-Schonlein purpura. PMID- 11865101 TI - Dialysis-associated acquired cystic kidney disease imitating autosomal dominant polycystic kidney disease in a patient receiving long-term peritoneal dialysis. PMID- 11865102 TI - Bone pain with scintigraphy suggestive of widespread metastases--do not forget phosphate. PMID- 11865103 TI - A 67-year-old kidney transplant patient with headache of uncertain origin. PMID- 11865104 TI - Migration of intra-vascular metallic stent into pulmonary artery. PMID- 11865105 TI - Transplant-associated inflammatory breast disease. PMID- 11865106 TI - A particularly private pain. PMID- 11865107 TI - Vascular access care and interventional radiology. PMID- 11865108 TI - Lupus nephritis in the new millennium. PMID- 11865109 TI - Dent's disease. PMID- 11865110 TI - Bartter syndrome type 3: an unusual cause of nephrolithiasis. PMID- 11865111 TI - Pulmonary hydatid disease associated with nephrotic syndrome in a paediatric patient. PMID- 11865112 TI - Aristolochic acid as a causative factor in a case of Chinese herbal nephropathy. PMID- 11865113 TI - Lupus nephritis: physicians' preparedness for communication of evidence-based therapy. PMID- 11865114 TI - Renal function, proteinuria and ACE-inhibitor therapy as determinants of plasma levels of endothelial markers. PMID- 11865115 TI - Accumulation of taurine in patients with renal failure. PMID- 11865116 TI - Can choice of dialyser membrane have a beneficial effect on HIV load in the HIV infected dialysis patient? PMID- 11865117 TI - Should low-dose methotrexate therapy be prescribed to dialysis patients? PMID- 11865118 TI - Pharmacokinetics and dosage adjustment of rifabutin in a haemodialysis patient. PMID- 11865119 TI - Heparin and dialysis: reasons to make a change? PMID- 11865120 TI - Intracerebral haemorrhage caused by cefazoline-induced hypoprothrombinaemia in a renal transplant recipient. PMID- 11865121 TI - Kidney donor evaluation with helical CT angiography. PMID- 11865123 TI - Attention deficit hyperactivity disorder and Tourette syndrome: what's the best treatment? PMID- 11865124 TI - Further evidence that for status epilepticus "one size fits all" doesn't fit. PMID- 11865125 TI - Opioids: a way to control central pain? PMID- 11865126 TI - Pharmaceutical industry support of the AAN Annual Meeting: policies and violation. PMID- 11865127 TI - Diagnostic criteria used in studies of reflex sympathetic dystrophy. AB - OBJECTIVE: Assessment of the diagnostic criteria of reflex sympathetic dystrophy (RSD) and evaluation of the impact of the introduction of the diagnostic criteria of complex regional pain syndrome (CRPS) on the international application of diagnostic criteria of RSD. METHODS: Randomized controlled trials and clinical investigations, published between January 1980 and June 2000, were evaluated with regard to the applied diagnostic criteria of RSD. RESULTS: One hundred seven studies were identified. Thirty-four of these studies were excluded because of inadequate reporting of diagnostic criteria. The 73 included studies were not homogeneous with regard to the diagnostic criteria because they applied many different aspects of sensory and autonomic features. Only 12% of the studies considered the presence of motor features, mostly vaguely described, as mandatory for the diagnosis RSD. Although 10 of the 23 studies published since the introduction of CRPS have applied this term, only 3 used the exact criteria without additions or other modifications. CONCLUSION: Diagnostic criteria sets of RSD focus on many different aspects of sensory and autonomic features that generally are described vaguely. This has not changed since the introduction of the CPRS criteria. These findings question whether the current criteria adequately define RSD. PMID- 11865128 TI - Treatment of ADHD in children with tics: a randomized controlled trial. AB - BACKGROUND: The treatment of children with attention deficit hyperactivity disorder (ADHD) and Tourette syndrome (TS) has been problematic because methylphenidate (MPH)--the most commonly used drug to treat ADHD--has been reported to worsen tics and because clonidine (CLON)--the most commonly prescribed alternative--has unproven efficacy. METHODS: The authors conducted a multicenter, randomized, double-blind clinical trial in which 136 children with ADHD and a chronic tic disorder were randomly administered CLON alone, MPH alone, combined CLON + MPH, or placebo (2 x 2 factorial design). Each subject participated for 16 weeks (weeks 1-4 CLON/placebo dose titration, weeks 5-8 added MPH/placebo dose titration, weeks 9-16 maintenance therapy). RESULTS: Thirty seven children were administered MPH alone, 34 were administered CLON alone, 33 were administered CLON + MPH, and 32 were administered placebo. For our primary outcome measure of ADHD (Conners Abbreviated Symptom Questionnaire--Teacher), significant improvement occurred for subjects assigned to CLON (p < 0.002) and those assigned to MPH (p < 0.003). Compared with placebo, the greatest benefit occurred with combined CLON + MPH (p < 0.0001). CLON appeared to be most helpful for impulsivity and hyperactivity; MPH appeared to be most helpful for inattention. The proportion of individual subjects reporting a worsening of tics as an adverse effect was no higher in those treated with MPH (20%) than those being administered CLON alone (26%) or placebo (22%). Compared with placebo, measured tic severity lessened in all active treatment groups in the following order: CLON + MPH, CLON alone, MPH alone. Sedation was common with CLON treatment (28% reported moderate or severe sedation), but otherwise the drugs were tolerated well, including absence of any evident cardiac toxicity. CONCLUSIONS: Methylphenidate and clonidine (particularly in combination) are effective for ADHD in children with comorbid tics. Prior recommendations to avoid methylphenidate in these children because of concerns of worsening tics are unsupported by this trial. PMID- 11865129 TI - Long-term mortality after a first episode of status epilepticus. AB - OBJECTIVE: To evaluate long-term mortality among people with status epilepticus (SE). METHODS: The authors performed a population-based retrospective cohort study to determine long-term mortality after SE. Between January 1, 1965, and December 31, 1984, all first episodes of SE receiving medical attention were ascertained through the Rochester Epidemiology Project Records-Linkage System. Cases surviving the first 30 days (n = 145) were followed until death or study termination (February 1996). RESULTS: At 10 years, cumulative mortality among 30 day survivors was 43%. The standardized mortality ratio (SMR) at 10 years was 2.8 (95% CI, 2.1-3.5). The mortality rate of those with idiopathic/cryptogenic SE was not increased (SMR = 1.1; 95% CI, 0.5-2.3). The following characteristics of SE increased long-term risk for mortality: SE > or = 24 hours in duration vs. SE < 2 hours (relative risk [RR] = 2.3; 95% CI, 1.1-5.1); acute symptomatic etiology vs idiopathic/cryptogenic etiology (RR = 2.2; 95% CI, 1.0-5.1) SE; myoclonic SE vs generalized convulsive SE (RR = 4.0; 95% CI, 1.3-13). CONCLUSION: Forty percent of subjects who survived the first 30 days after an incident episode of SE die within the next 10 years. The long-term mortality rate was threefold that of the general population over the same time period. The long-term mortality rate at 10 years was worse for those with myoclonic SE, for those who presented with SE lasting more than 24 hours, and for those with acute symptomatic SE. The long term mortality rate was not altered in those with idiopathic/cryptogenic SE. We conclude that SE alone does not modify long-term mortality. PMID- 11865130 TI - Seizure-associated brain injury in term newborns with perinatal asphyxia. AB - BACKGROUND: There is controversy over whether seizures, the most common manifestation of neonatal brain injury, may themselves damage the developing brain. OBJECTIVE: To determine if neonatal seizures are independently associated with brain injury in newborns with perinatal asphyxia. METHODS: Ninety term neonates were studied with MRI and single-voxel (1)H-MRS on median day of life 6 (range 1 to 13 days). The severity of MR abnormality in the (1)H-MRS regions of interest was scored using a validated scale. Seizure severity was scored based on seizure frequency and duration, EEG findings, and anticonvulsant administration. Multivariable linear regression tested the independent association of seizure severity with impaired cerebral metabolism measured by lactate/choline and compromised neuronal integrity measured by N-acetylaspartate/choline in both regions. RESULTS: Clinical seizures occurred in 33 of 90 infants (37%). Seizure severity was associated with increased lactate/choline in both the intervascular boundary zone (p < 0.001) and the basal nuclei (p = 0.011) when controlling for potential confounders of MRI abnormalities and amount of resuscitation at birth. Each increase in seizure score was independently associated with a 21% increase in lactate/choline in the intervascular boundary zone (95% CI, 5.1-38.2%) and a 15% increase in the basal nuclei (95% CI, 0.1-31.7%). Seizure severity was independently associated with diminished N-acetylaspartate/choline in the intervascular boundary zone (p = 0.034). CONCLUSION: The severity of seizures in human newborns with perinatal asphyxia is independently associated with brain injury and is not limited to structural damage detectable by MRI. PMID- 11865131 TI - Enzyme-inducing antiepileptic drugs in pregnancy and the risk of bleeding in the neonate. AB - BACKGROUND: Case reports suggest that maternal hepatic enzyme-inducing antiepileptic drugs (AED) increase the risk for neonatal bleeding. Antenatal administration of vitamin K(1) to mothers using these drugs therefore is widely recommended. There are, however, no studies on the incidence of this complication. OBJECTIVE: To assess the occurrence of bleeding complications in newborns exposed to maternal enzyme-inducing AED in utero. METHODS: The authors prospectively followed 662 pregnancies in women with epilepsy who used enzyme inducing AED. Of the 667 neonates, 463 were exposed to carbamazepine, 212 to phenytoin, 44 to phenobarbital, 11 to primidone, and 7 to oxcarbazepine. The control subjects were 1,324 nonepileptic pregnancies (1,334 neonates) matched for maternal age, parity, number of fetuses, and delivery date. None of the mothers received vitamin K(1) during pregnancy, but all infants received 1 mg vitamin K(1) intramuscularly at birth. RESULTS: A bleeding complication was observed in five (0.7%) of the offspring exposed to maternal enzyme-inducing AED and in five (0.4%) control subjects (p = 0.3). After logistic regression analysis was performed, bleeding was associated with birth at <32 weeks of gestation (adjusted OR = 13; 95% CI = 2.7 to 64) and alcohol abuse (adjusted OR = 17; 95% CI = 1.8 to 162) but not with exposure to enzyme-inducing AED (adjusted OR = 1.1; 95% CI = 0.3 to 4.6; p = 0.8). CONCLUSIONS: These data do not support the hypothesis that maternal enzyme-inducing AED increase the risk for bleeding in the offspring. Antenatal administration of vitamin K to these mothers may still be needed in selected cases. PMID- 11865132 TI - Effects of IV morphine in central pain: a randomized placebo-controlled study. AB - OBJECTIVE: To investigate the effects of IV morphine on central pain syndromes through quantitative sensory testing and to assess the long-term benefit of oral morphine. METHODS: After an initial open titration phase aiming to determine the maximal tolerated dosage of IV morphine, the efficacy of morphine infusion (9-30 mg; mean dosage, 16 mg) was assessed in a double-blind, placebo-controlled and crossover fashion in 15 patients with poststroke- (6 patients) or spinal cord injury- (9 patients) related pain. All of the patients subsequently received sustained oral morphine. RESULTS: Morphine significantly reduced the intensity of brush-induced allodynia but had no effect on other evoked pains (i.e., static mechanical and thermal allodynia/hyperalgesia). The effects of morphine on ongoing pain were not significantly different from those of the placebo, but 7 patients (46%) responded to morphine. There was a correlation between the effects of morphine on spontaneous pain and the decrease of the responses to suprathreshold thermal stimuli on the nonpainful contralateral side, suggesting that these effects were related to the general antinociceptive activity of the drug. The effects of IV morphine were correlated with those of oral morphine at 1 month, but only 3 patients (20%) were still taking morphine after 1 year. CONCLUSIONS: IV morphine induces analgesic effects on some components of central neuropathic pain syndromes, but only a minority of patients may benefit from long term opioid treatment. PMID- 11865133 TI - COMT genotype and effectiveness of entacapone in patients with fluctuating Parkinson's disease. AB - OBJECTIVE: To investigate the relationship between the catechol-O methyltransferase (COMT) genotype and the therapeutic efficacy of entacapone. METHODS: The efficacy of 2 months of entacapone treatment in 65 patients with PD with end-of-dose deterioration was studied. The efficacy of entacapone was assessed using the Unified Parkinson's Disease Rating Scale (UPDRS) score, the daily levodopa dosage, and the patients' diary card. RESULTS: Thirty-six patients (55.4%) had a high-activity COMT gene (COMT(HH)), 22 (33.8%) had an intermediate activity COMT gene (COMT(HL)), and 7 patients (10.8%) had a low-activity COMT gene (COMT(LL)). Two months of entacapone treatment resulted in a significant increase in "on" time, a reduction in "off" time, and a reduction in the total UPDRS score, but these results were independent of the COMT genotype of the patient. There was no significant difference in the frequency or severity of dyskinesias between the patients with different COMT genotypes. CONCLUSION: The COMT genotype seems to be a minor factor in judging the beneficial effects of entacapone administration. PMID- 11865134 TI - A multi-incident, Old-Order Amish family with PD. AB - BACKGROUND: PD is largely a sporadic condition of unknown etiology, but specific inherited mutations are a cause of PD. OBJECTIVE: To describe a large, multi incident Amish pedigree with PD. METHODS: Case ascertainment, calculation of population prevalence, and calculation of kinship coefficients (a measure of relatedness between two individuals) for affecteds and subjects in a large kindred with PD were conducted. Sequencing of genes with known mutations sufficient to cause PD and marker-by-marker haplotype analysis in chromosomal regions flanking previously described genes with known mutations were performed. RESULTS: The authors have examined 113 members of this pedigree and classified 67 as normal (no evidence of PD), 17 as clinically definite PD, 6 as clinically probable PD, and 23 as clinically possible PD. The mean age at onset of the clinically definite subjects was 56.7 years. The phenotype in this family is typical of idiopathic PD, including rest tremor, rigidity, bradykinesia, postural instability, and response to levodopa. In addition, dementia occurred in six of the clinically definite subjects, and many subjects experienced levodopa-related motor complications including wearing off and dopa-induced dyskinesias. In the index Amish community, a minimum prevalence of PD in the population 40 years and older of 552/100,000 was calculated. The mean kinship coefficient in the subjects with PD and those with PD by history (0.036) was higher (p = 0.007) than in a group of age-matched normal Amish control subjects (0.016), providing evidence that PD is inherited in this family. Sequence analysis did not detect any mutations in known PD genes. No single haplotype cosegregates with the disease in any of the chromosomal regions previously found to be linked to PD, and no marker in these regions exhibits increased homozygosity among definite PD cases. CONCLUSIONS: PD in this community is more common than in the general population, and this increased prevalence may be due in part to a novel gene(s). PMID- 11865135 TI - Diffusion-weighted MRI differentiates the Parkinson variant of multiple system atrophy from PD. AB - OBJECTIVE AND BACKGROUND: Routine MRI as well as MR volumetry and MRS have been shown to contribute to the differential diagnosis of the Parkinson variant of multiple system atrophy (MSA-P) and PD. However, it is currently unknown whether diffusion-weighted imaging (DWI) discriminates these disorders. METHODS: Ten patients with MSA-P (mean age, 64 years) were studied, 11 with PD (mean age, 64 years), and seven healthy volunteers (mean age, 59 years) matched for age and disease duration. Regional apparent diffusion coefficients (rADC) were determined in different brain regions including basal ganglia, gray matter, white matter, substantia nigra, and pons. RESULTS: Patients with MSA-P had higher putaminal rADC (median 0.791 x 10(3)/mm(2)/s) than both patients with PD (median 0.698 x 10(3)/mm(2)/s, p < 0.001) and healthy volunteers (median 0.727 x 10(3)/mm(2)/s, p < 0.001). There were no significant differences in putaminal rADC between patients with PD and healthy volunteers. Moreover, none of the putaminal rADC values in the PD and control group surpassed the lowest value in the MSA-P group. There were no significant group differences in the rADC values in other brain regions such as pons, substantia nigra, globus pallidus, caudate nucleus, thalamus, or gray and white matter. Putaminal rADC values correlated significantly with Unified PD Rating Scale OFF scores in patients with MSA as measured by the Spearman rank test. CONCLUSION: DWI, even if measured in the slice direction only, is able to discriminate MSA-P and both patients with PD and healthy volunteers on the basis of putaminal rADC values. The increased putaminal rADC values in Parkinson variant of multiple system atrophy are likely to reflect ongoing striatal degeneration, whereas most neuropathologic studies reveal intact striatum in PD. Diffusion-weighted imaging may represent a useful diagnostic tool that can provide additional support for a diagnosis of Parkinson variant of multiple system atrophy. PMID- 11865136 TI - Smoking and Parkinson's disease in twins. AB - OBJECTIVE: To test the hypothesis that cigarette smoking protects against the development of PD. BACKGROUND: Smoking has been inversely associated with PD in many studies, but whether this reflects a biologic effect on the underlying disease process or merely confounding or selection bias remains uncertain. METHODS: The authors compared smoking histories in male twin pairs identified from the National Academy of Sciences--National Research Council World War II Veteran Twins Cohort. The amount of cigarettes smoked (in pack-years) was collected until the time of PD onset in the affected twin or until the time of death for the unaffected twin, whichever came first. Differences in pack-years smoked until PD onset and until 10 and 20 years before onset were compared using paired t-tests. Comparisons were made overall and stratified by zygosity and concordance for PD. To assess the role of shared environment, correlation for smoking behaviors was compared between pairs concordant and discordant for PD. RESULTS: Detailed smoking histories were available for 113 twin pairs in which at least one twin had PD (discordant pairs: 43 monozygotic [MZ], 50 dizygotic [DZ]; concordant pairs: 10 MZ, 10 DZ). Within-pair correlation for ever smoking was high in MZ pairs (phi = 0.47, p = 0.001) but not in DZ pairs (phi = 0.007, p = 0.96). In 33 discordant MZ pairs and 39 discordant DZ pairs in which at least one twin had smoked, the twins without PD smoked more than their brothers smoked (32.5 vs. 22.7 pack-years, p = 0.026). This was more marked in the MZ pairs (37.1 vs. 25.3 pack-years, p = 0.077) than in the DZ pairs (28.6 vs. 20.5 pack-years, p = 0.17). A similar relationship was seen when smoking dose was calculated only until 10 years before PD onset, suggesting that the lower dose of smoking in the twin with PD was not the result of early, undiagnosed disease. CONCLUSION: Within twin pairs, risk of PD is inversely correlated with the dose (in pack-years) of cigarette smoking. This effect is most pronounced in MZ twins, despite the high correlation for smoking. Because MZ twins are genetically identical and are similar behaviorally, this difference is unlikely to result from either genetic factors or environmental confounders. These results are compatible with a true biologic protective effect of cigarette smoking. PMID- 11865137 TI - Increase of cdk5 is related to neurofibrillary pathology in progressive supranuclear palsy. AB - BACKGROUND: Progressive supranuclear palsy (PSP) is characterized by a pure neurofibrillary tau pathology involving mainly basal ganglia and brainstem nuclei. In addition to a haplotype of the tau gene potentially favoring tau aggregation, lipoperoxidation has been shown to be associated with PSP tau pathology. OBJECTIVE: To analyze cdk5/p35 complex, a kinase that regulates neurite outgrowth, as a potential cellular mechanism underlying tau phosphorylation in brain tissues from PSP and control cases and comparatively in cerebral cortex from subjects with AD. METHODS: Cdk5/p35 protein levels and distribution were evaluated by immunoblotting and immunocytochemistry in brain regions from seven PSP, six AD, and seven control cases, with similar postmortem intervals. RESULTS: Total cdk5 protein levels were significantly increased by more than threefold in PSP tissue and were augmented in PSP neurons, codistributed with tau immunoreactivity. P35, the regulatory subunit of cdk5, was degraded by postmortem proteolysis to the same extent in PSP, AD, and control tissues. CONCLUSIONS: The proteolysis in vivo of p35, the regulatory subunit of the kinase, is not ascertainable because it is masked by its postmortem degradation. The study, however, indicates that in PSP, the alteration of cdk5 is different from that described in AD and suggests that the absence of amyloid beta protein deposition may account for the different pathways responsible for the same kinase activation. PMID- 11865138 TI - Novel mutations in collagen VI genes: expansion of the Bethlem myopathy phenotype. AB - OBJECTIVE: To investigate the molecular basis of autosomal dominant limb-girdle muscular dystrophy (AD-LGMD) in three large new families. METHODS AND RESULTS: Genome-wide linkage was performed to show that the causative gene in all three families localized to chromosome 21q22.3 (Zmax = 10.3; theta = 0). This region contained the collagen VI alpha1 and alpha2 genes, which have been previously shown to harbor mutations causing a relatively mild congenital myopathy with contractures (Bethlem myopathy). Screening of the collagen VI alpha1 and alpha2 genes revealed novel, causative mutations in each family (COL6A1-K121R, G341D; COL6A2-D620N); two of these mutations were in novel regions of the proteins not previously associated with disease. Collagen VI is a ubiquitously expressed component of connective tissue; however, both limb-girdle muscular dystrophy and Bethlem myopathy patients show symptoms restricted to skeletal muscle. To address the muscle-specific symptoms resulting from collagen VI mutations, the authors studied three patient muscle biopsies at the molecular level (protein expression). A marked reduction of laminin beta1 protein in the myofiber basal lamina in all biopsies was found, although this protein was expressed normally in the neighboring capillary basal laminae. CONCLUSIONS: The authors' studies widen the clinical spectrum of Bethlem myopathy and suggest collagen VI etiology should be investigated in dominant limb-girdle muscular dystrophy. The authors hypothesize that collagen VI mutations lead to muscle-specific defects of the basal lamina, and may explain the muscle-specific symptoms of Bethlem and limb girdle muscular dystrophy patients with collagen VI mutations. PMID- 11865139 TI - Three novel COLQ mutations and variation of phenotypic expressivity due to G240X. AB - OBJECTIVE: To determine the molecular basis and consequences of endplate (EP) acetylcholinesterase (AChE) deficiency. BACKGROUND: The EP species AChE is an asymmetric enzyme consisting of a tail subunit composed of three collagenic strands (ColQ), each attached to a tetramer of catalytic subunits. The tail subunit is essential for insertion of AChE into the synaptic basal lamina. Human EP AChE deficiency is caused by mutations in COLQ. The authors report three novel COLQ mutations in eight kinships. METHODS: Immunocytochemistry, electron microscopy, microelectrode recordings, mutation analysis, and expression studies in COS cells were employed. RESULTS: Two mutations (275insC and Q211X) were heterozygous in one patient. EP studies in this patient revealed no EP AChE, small nerve terminals, reduced presynaptic membrane length, as well as abnormally low-evoked quantal release. The third mutation (G240X) was homozygous in six Palestinian Arab families of the same tribe and in an Iraqi Jewish patient. Expression studies of the three mutations in COS cells indicate that each abrogates formation of insertion competent asymmetric AChE. Although the three mutations have identical predicted consequences at the EP, their phenotypic expressivity varies as regards age at onset, rate of progression, and severity of symptoms. CONCLUSIONS: 1) After mutations in the AChR epsilon subunit, mutations in COLQ are emerging as second most common cause of congenital myasthenic syndromes. 2) A founder effect is likely for G240X in the Palestinian Arab families. 3) That mutations predicting total absence of AChE from the EP have variable phenotypic expressivity suggests that modifying genes or environmental factors can partially compensate for EP AChE deficiency. PMID- 11865140 TI - Infections and course of disease in mild forms of Guillain-Barre syndrome. AB - OBJECTIVE: Twenty-eight percent of patients with the Guillain-Barre syndrome remain able to walk unaided. Studying patients with the mild form of Guillain Barre syndrome can further contribute to knowledge of the spectrum of the syndrome and explore whether this subgroup may need treatment with IV immunoglobulin. METHODS: Patients fulfilling the National Institute of Neurologic and Communicative Disorders and Stroke criteria for Guillain--Barre syndrome were included in a nationwide survey over a 2-year period. Clinical characteristics and serum samples were collected prospectively. In addition, a questionnaire was completed concerning the course and outcome of the disease. RESULTS: A total of 139 patients were included. Nineteen of the patients (14%) included were mildly affected, and 120 (86%) were severely affected. Infections with Epstein-Barr virus were found more frequently in mildly affected patients (p = 0.02). Antiganglioside antibodies were less frequently found in the mildly affected patients (p = 0.03). The degree of severity of the disease between mildly and severely affected patients was different on the day of admission (p < 0.01). Thereafter, the groups showed a remarkably similar rate of progression. Thirty eight percent of mildly affected patients report problems in hand function and an inability to run at 3 and 6 months (all women, p = 0.02). CONCLUSION: The difference in severity of Guillain--Barre syndrome seems to be determined in an early phase of the disease. Preceding infections and antiganglioside antibodies may influence the initial immune attack, determining the severity of the disease. The presence of residual signs in patients with mild disease may advocate the use of early treatment in mildly affected patients. PMID- 11865141 TI - Axonal multifocal motor neuropathy without conduction block or other features of demyelination. AB - BACKGROUND: Conduction block is considered an essential finding for the distinction between motor neuropathies and lower motor neuron disorders. Only a small number of reports describe patients with multifocal motor neuropathies who lack overt conduction block, although in these cases other features of demyelination still suggest the presence of a demyelinating disorder. In contrast, a purely axonal multifocal motor neuropathy has not been described. METHODS: This report describes nine patients with slowly or nonprogressive multifocal motor neuropathies who had purely axonal electrodiagnostic features. RESULTS: GM1 antibodies titers were normal in all nine cases. Six patients were treated with either prednisone or IV immunoglobulin and three showed convincing improvement. CONCLUSIONS: These findings suggest an immune-mediated motor neuropathy with axonal electrophysiologic features that appears to be distinct from both multifocal motor neuropathy and established motor neuron disorders. PMID- 11865142 TI - Ptosis in patients with hemispheric strokes. AB - BACKGROUND: Cerebral ptosis is considered rare; it has been reported with unilateral, usually right hemispheric lesions. However, the frequency of cerebral ptosis in patients with strokes has not received systematic study. OBJECTIVE: To determine the frequency of ptosis in patients with acute hemispheric stroke and to identify stroke features associated with ptosis. METHODS: Eyelid function was studied in 64 consecutive patients with acute hemispheric stroke and 40 age matched subjects with no known neurologic disease. All underwent comprehensive neuro-ophthalmologic and general neurologic examination within 48 hours of admission, including measurement of palpebral fissures, marginal reflex distance, and range of upper lid movement. Only patients who could cooperate with eyelid testing were included. Brain CT scans were obtained for all patients who had had strokes. RESULTS: Twenty-four (37.5%) of the patients with strokes had neurogenic ptosis, which was bilateral in 10 and unilateral in 14. None of the control subjects had neurogenic ptosis. All patients with strokes with ptosis had a hemiparesis. Rightward gaze deviation and upgaze paresis were more common (p < 0.05) in the patients with ptosis compared with others who had had strokes. CT evidence of right-sided hemispheric cortical infarction was more common in patients with strokes with ptosis (p < 0.05). In five patients with large hemispheric infarction, complete bilateral or asymmetric ptosis was the first sign of imminent herniation, preceding pupillary dilation and ocular motor deficits. CONCLUSIONS: Ptosis occurs frequently in patients with hemispheric strokes, especially in association with right hemispheric lesions. Complete bilateral ptosis is usually caused by large infarctions and may be a premonitory sign of an impending herniation. PMID- 11865143 TI - Molecular signatures of brain injury after intracerebral hemorrhage. AB - BACKGROUND: The mechanisms of cellular death in the tissue surrounding an intracerebral hemorrhage (ICH) are not defined. OBJECTIVE: To investigate the relationship of markers of excitotoxicity and inflammation to brain injury after ICH. METHODS: A total of 124 consecutive patients with spontaneous ICH admitted within 24 hours of stroke onset were prospectively investigated. The volumes of the initial ICH, peripheral edema on days 3 to 4, and the residual cavity at 3 months were measured on CT scan. Glutamate, cytokines, and adhesion molecules were measured in blood samples obtained on admission. Stroke severity and neurologic outcome were evaluated with the Canadian Stroke Scale. RESULTS: Poor neurologic outcome at 3 months (Canadian Stroke Scale < 7) was observed in 53 patients (43%). Stroke severity and glutamate concentrations (by each increment of 10 micromol/L, odds ratio 1.23; 95% CI 1.09 to 1.41), but not the initial volume of ICH, were independent predictors of poor outcome. In the multiple linear regression analyses, tumor necrosis factor-alpha concentration was correlated (r = 0.83, p < 0.0001) with the volume of perihematoma edema, and glutamate concentrations were correlated (r = 0.78, p < 0.0001) with the volume of the residual cavity. These same results were observed when lobar (n = 58) and deep (n = 66) ICH were analyzed separately. CONCLUSIONS: High plasma levels of proinflammatory molecules within 24 hours of intracerebral hemorrhage onset are correlated with the magnitude of the subsequent perihematoma brain edema, whereas poor neurologic outcome and the volume of the residual cavity are related to increased plasma glutamate concentrations. PMID- 11865144 TI - Neurophysiological correlates of age-related changes in human motor function. AB - BACKGROUND: There are well-defined and characteristic age-related deficits in motor abilities that may reflect structural and chemical changes in the aging brain. OBJECTIVE: To delineate age-related changes in the physiology of brain systems subserving simple motor behavior. METHODS: Ten strongly right-handed young (<35 years of age) and 12 strongly right-handed elderly (>50 years of age) subjects with no evidence of cognitive or motor deficits participated in the study. Whole-brain functional imaging was performed on a 1.5-T MRI scanner using a spiral pulse sequence while the subjects performed a visually paced "button press" motor task with their dominant right hand alternating with a rest state. RESULTS: Although the groups did not differ in accuracy, there was an increase in reaction time in the elderly subjects (mean score plus minus SD, young subjects = 547 +/- 97 ms, elderly subjects = 794 +/- 280 ms, p < 0.03). There was a greater extent of activation in the contralateral sensorimotor cortex, lateral premotor area, supplementary motor area, and ipsilateral cerebellum in the elderly subjects relative to the young subjects (p < 0.001). Additional areas of activation, absent in the young subjects, were seen in the ipsilateral sensorimotor cortex, putamen (left > right), and contralateral cerebellum of the elderly subjects. CONCLUSIONS: The results of this study show that elderly subjects recruit additional cortical and subcortical areas even for the performance of a simple motor task. These changes may represent compensatory mechanisms invoked by the aging brain, such as reorganization and redistribution of functional networks to compensate for age-related structural and neurochemical changes. PMID- 11865145 TI - Ictal heart rate differentiates epileptic from non-epileptic seizures. AB - Video-EKG-EEG recordings of 67 epileptic seizures and 38 psychogenic nonepileptic seizures were reviewed. Ictal heart rate (HR) was higher during and after epileptic seizures for both convulsive and nonconvulsive spells (p < 0.01). When analyzing quiet staring spells separately, it was possible to identify which spells were epileptic in origin with a positive predictive value of 97% by using a cutoff of a > or =30% increase in HR over baseline. It was concluded that documenting an increase in HR during a spell of unresponsiveness usually can distinguish between epileptic and psychogenic causes. PMID- 11865146 TI - Functional changes in temporal lobe activity during transient global amnesia. AB - The integrity of temporal lobe activity during and after recovery from transient global amnesia (TGA) was assessed in a case study using functional MRI. TGA was associated with scene-encoding deficits in a temporolimbic circuit that recovered over time. Frontoparietal areas recruited during the amnesic state may signify a compensatory reliance on visuospatial or working memory strategies. Reduction of extrastriate cortex responses over repeated testing sessions possibly indicates intact visual priming in TGA. PMID- 11865147 TI - Ipsilateral thalamic MRI abnormality in an epilepsy patient. AB - In a 19-year-old patient with status epilepticus arising in the right parietal neocortex, unenhanced ictal MRI showed abnormalities mainly in the right cerebral cortex, contralateral cerebellum, and ipsilateral thalamus. The thalamus is considered a key site of functional abnormality in this patient. PMID- 11865148 TI - Familial nocturnal facio-mandibular myoclonus mimicking sleep bruxism. AB - A mother and son presented with a multi-decade history of nocturnal tongue biting and bleeding. In both patients, video polysomnographic recordings documented bursts of electromyographic activity originating in the masseter and spreading to orbicularis oris and oculi muscles, present only during sleep. Faciomandibular myoclonic activity during sleep mimics sleep bruxism and may be familial. PMID- 11865149 TI - Serial MRI findings in a case of primary lateral sclerosis. AB - A patient with primary lateral sclerosis (PLS) underwent MRI 3 times over 8.5 years. These images demonstrate progressive atrophy of the premotor, parietal, and primary sensorimotor cortex, with sparing of the temporal lobe, occipital lobe, and cerebellum. The pattern of cerebral atrophy was clearly restricted but was more extensive than expected from sensorimotor cortex pyramidal cell loss described in literature autopsy reports of PLS. Cortical motor system degeneration in PLS, whether primary or secondary, may be more extensive than previously believed. PMID- 11865150 TI - Not paralysis, but dystonia causes stridor in multiple system atrophy. AB - Electromyography (EMG) was performed in 10 patients with multiple system atrophy, laryngeal or pharyngeal symptoms, or both. In patients with stridor, EMG during quiet breathing revealed persistent tonic activity in both abductor and adductor vocal cord muscles. In patients with dysphagia, the cricopharyngeal muscle showed persistent EMG activity throughout all phases of swallowing. Botulinum toxin injection into the adductor muscle determined subjective improvement and reduced tonic EMG activity. Therefore, the cause of stridor in multiple system atrophy is dystonia of the vocal cords. PMID- 11865151 TI - Higher lipoprotein (a) levels in atherothrombotic than lacunar ischemic cerebrovascular disease. AB - To investigate the role of plasma lipid abnormalities in ischemic cerebrovascular disease related to primary vessel disease, the authors assess lipid profiles in a hospital-based cohort of 202 consecutive patients with atherothrombotic or lacunar stroke subtypes. Lipoprotein (a) was the unique lipid parameter that differs between these two subtypes being its value twofold higher in patients with atherothrombotic than in lacunar stroke. This suggests that lipoprotein (a) promotes large vessel atheromatosis rather than small vessel arteriolosclerosis and favors thrombosis on atheromatous plaques by suppressing local fibrinolysis. PMID- 11865152 TI - Diffusion abnormalities in patients with Wernicke encephalopathy. AB - Diffusion-weighted imaging (DWI) can help to diagnose acute ischemic stroke. Other nonischemic disorders may show abnormal signals with DWI. The authors report two cases of Wernicke encephalopathy with DWI signal changes in characteristic midline locations, one with reduction in apparent diffusion constant and one without. DWI abnormalities may suggest early thiamine deficiency and are useful in diagnosing Wernicke encephalopathy. PMID- 11865153 TI - Association and linkage of juvenile MS with HLA-DR2(15) in Russians. AB - The aim of this study was to determine the role of the HLA-DRB1 gene [6p21] in susceptibility to juvenile MS (JMS) (age at onset < or =15 years) of children of Russian descent. Association of DR2(15) with JMS has been found by the comparison of patients with JMS with both unrelated and affected family-based healthy controls. The linkage of DR2(15) with JMS was shown by transmission disequilibrium test. There were no significant differences in the frequencies of DRB1 alleles and genotypes between 56 patients with JMS and 234 patients with MS with age at onset > or =16 years. PMID- 11865154 TI - Cervical disc prolapse with cord compression presenting with choreoathetosis and dystonia. PMID- 11865155 TI - Zolpidem improves dystonia in "Lubag" or X-linked dystonia-parkinsonism syndrome. PMID- 11865156 TI - Disopyramide-induced neuropathy. PMID- 11865157 TI - A polymorphism in the cystatin C promoter region is not associated with an increased risk of AD. PMID- 11865158 TI - Cyclist's doping associated with cerebral sinus thrombosis. PMID- 11865159 TI - Effect of food on absorption of Dilantin Kapseals and Mylan extended phenytoin sodium capsules. PMID- 11865160 TI - The cost of delaying treatment in multiple sclerosis: what is lost is not regained. PMID- 11865161 TI - Headache characteristics in patients after migrainous stroke. PMID- 11865162 TI - Effects of internal globus pallidus stimulation on motor cortex excitability. PMID- 11865163 TI - Mitochondrial myopathy, parkinsonism and multiple mtDNA deletions in a Sephardic Jewish family. PMID- 11865164 TI - Prevalence of Alzheimer's disease in very elderly people: a prospective neuropathological study. PMID- 11865167 TI - What do patients really want to know? PMID- 11865168 TI - Cerebral venous sinus thrombosis. PMID- 11865169 TI - Survival of unselected stroke patients in a stroke unit compared with conventional care. AB - BACKGROUND: Recent meta-analyses have reported a beneficial effect of stroke units compared with traditional care, both on patient survival and on dependency after one year. AIM: To determine whether these results can be reproduced outside a clinical trial setting. SETTING: A medium-sized general hospital. METHODS: From 1993 to 1998, all patients aged >60 years with suspected acute stroke were allocated either to a stroke unit or general medical wards according to date of birth (day of the month). Patients were identified retrospectively, using a discharge diagnosis of ICD-9 codes 431, 434 and 436. We assessed 30-day and 1 year survival. RESULTS: In the stroke unit, 926/1128 patients survived at 30 days, vs. 905/1141 in the general medical wards (p=0.08). Beyond the first 30 days, there was no difference in survival (p=0.27). Under Cox regression analysis, there was a 20% reduction in mortality in the stroke unit after 30 days compared with the general medical wards (RR 0.80, p=0.02) after adjusting for age, gender, stroke type and season of the year. DISCUSSION: In this, the largest single-centre study to evaluate the survival benefit of a stroke unit, survival at 30 days was increased, although not significantly so. Survival at one year was unchanged. PMID- 11865170 TI - Management of hyperemesis gravidarum: the importance of weight loss as a criterion for steroid therapy. AB - BACKGROUND: Although the effectiveness of prednisolone therapy for severe hyperemesis gravidarum has been demonstrated, there is no consensus on how to assess severity to justify such treatment, nor any information on whether such therapy affects birth weight. AIM: To document the effect of prednisolone therapy in women with defined severity of hyperemesis gravidarum. DESIGN: Single centre, observational study of 30 consecutive pregnancies complicated by hyperemesis and weight loss of >5% of pre-pregnant weight between April 1995 and July 2000. Comparison of birth weight with a contemporaneous control series of women admitted with hyperemesis that was judged insufficiently severe to require steroids. RESULTS: Treatment with prednisolone 10 mg tid rapid resolved nausea and vomiting, allowing discharge in 3 (range 1-6.5) days. Steroid therapy, which was reduced in a stepwise manner, was discontinued at a median gestation of 20 weeks. Maternal weight gain in pregnancy was restored to normal. Median birth weight in the severe, steroid-treated group was 3.33 (range 2.80-3.27) kg vs. 3.27 (range 3.04-3.53) kg in the less severe group. CONCLUSION: Weight loss >5% served as a criterion to define a subset of women with severe hyperemesis gravidarum. In these women, steroid therapy was uniformly successful resulting in the prompt resolution of symptoms. Steroid therapy did not affect birth weight. PMID- 11865171 TI - Abnormal endothelin B receptor vasomotor responses in patients with Hirschsprung's disease. AB - BACKGROUND: Hirschsprung's disease is associated with defects in the endothelin-3 and endothelin B receptor genes. AIM: To assess the in vivo vasomotor responses to endothelin B receptor stimulation in patients with Hirschsprung's disease. METHODS: Forearm blood flow was measured using venous occlusion plethysmography in 10 patients with Hirschsprung's disease and 10 matched healthy controls during intra-brachial infusion of the highly selective endothelin B receptor agonist, sarafotoxin S6c. To simulate endothelin B receptor dysfunction, sarafotoxin S6c was co-infused with the highly selective endothelin B receptor antagonist, BQ 788, in six of the healthy controls. RESULTS: Sarafotoxin S6c caused a brief initial vasodilatation followed by a slow-onset, sustained vasoconstriction (p<0.001). Compared to control subjects, patients with Hirschsprung's disease had a substantial impairment of the initial vasodilatation whilst producing a more pronounced subsequent vasoconstriction (p<0.001). In healthy controls, co infusion of BQ-788 and sarafotoxin S6c caused a similar pattern of responses to those obtained in patients with Hirschsprung's disease: abolition of the initial vasodilatation and augmentation of subsequent vasoconstriction (p<0.001). DISCUSSION: In the majority of patients with Hirschsprung's disease, there is a functional defect of the vascular endothelin B receptor. PMID- 11865172 TI - Abnormal low-density lipoprotein subfraction profile in patients with untreated hypertension. AB - BACKGROUND: Low-density lipoprotein (LDL) consists of a heterogeneous group of particles of varying size and electrophoretic mobility. A predominance of small, more mobile particles is a risk factor for cardiovascular disease. AIM: To investigate the hypothesis that untreated patients with essential hypertension in the absence of vascular disease may exhibit abnormalities of LDL subfractions. SETTING: Specialist hypertension clinic. DESIGN: Cross-sectional study. METHODS: Following disc polyacrylamide gel electrophoresis, the mean (LDL locus) and heterogeneity (LDL spread) of mobility was recorded in 41 patients (mean age 52.6 years, 24 men) presenting with untreated essential hypertension (in the absence of vascular disease or diabetes mellitus) and in 45 healthy controls (age 56.9 years, 22 men) recruited from primary-care lists. RESULTS: Although there were no significant differences in total, low- or high-density lipoprotein cholesterol concentrations, LDL locus was significantly greater in the hypertensive group: mean (95%CI) 36.7 (35.7-37.6) vs. 34.8 (34.1-35.5), p=0.002. LDL locus was significantly elevated even in hypertensives with triglyceride concentrations below the median (<1.25 mmol/l). LDL spread was also greater in the hypertensive group, but not significantly: 5.6 (5.2-6.0) vs. 5.5 (5.3-5.8), p=0.10. DISCUSSION: Hypertensive patients have a preponderance of smaller LDL subfractions. This dyslipidaemia is not readily detected by conventional lipid assays. PMID- 11865173 TI - Complement activation in postpartum thyroiditis. AB - BACKGROUND: Postpartum thyroid dysfunction (PPTD) develops in 50% of pregnant women who have raised levels of circulating thyroid peroxidase autoantibodies (TPOAb) at booking. Although these antibodies are able to activate the complement cascade in vitro, it is not known whether complement activation plays any role in the pathogenesis of this disease. AIM: To investigate potential and actual activation of the complement system in women with postpartum thyroiditis. DESIGN: Complement activation was monitored on a weekly basis in 24 postpartum women who had raised TPOAb at 16 weeks gestation, attending an antenatal clinic in Mid Glamorgan, Wales. METHODS: ELISA procedures were used to measure both in-vitro complement C3 activation by TPOAb and circulating terminal complement complexes (TCC) in serum. RESULTS: Higher levels of bioactive TPOAb activity were seen in women who developed PPTD when compared to those who did not. However, TCC remained undetectable in serum throughout the period of study. CONCLUSIONS: In PPTD, despite the presence of circulating bioactive TPOAbs, the extent of complement activation is inadequate to cause detectable increases in peripheral blood TCC, suggesting that the complement system may not play a major role in PPTD pathogenesis. PMID- 11865174 TI - Medicines management: a sour taste. PMID- 11865175 TI - Accidental occupational exposure to CNS-toxic substances in bank employees. PMID- 11865176 TI - Occult papillary thyroid carcinoma presenting as extrathyroidal solitary neck cyst. PMID- 11865177 TI - Pericardial disease associated with Grave's thyrotoxicosis. PMID- 11865178 TI - Septic discitis presenting following intravenous cannulation. PMID- 11865179 TI - Get ahead! PMID- 11865180 TI - Darwin's dangerous idea. PMID- 11865181 TI - Signal transduction by MAP kinases: regulation by phosphorylation-dependent switches. AB - The kinases of mitogen-activated protein (MAP) kinase cascades transmit signals through sequential phosphorylation and activation of the enzymes. However, recent evidence indicates that protein-protein interactions between the kinases themselves or with substrates or other components are also a critical means of regulation. Whitmarsh and Davis summarize these findings with emphasis on new evidence from yeast that, when phosphorylated, a MAP kinase kinase actually switches from a negative regulator that binds to and inhibits its target MAP kinase to a positive regulator of that same enzyme. PMID- 11865182 TI - Growth factors and gene expression: fresh insights from arrays. AB - Gene array technology allows researchers to evaluate patterns of gene expression at a genome-wide level. Two recent papers have applied this powerful technique to characterize how gene expression is changed in response to growth factors and mitogens. The studies focus on two important questions concerning specificity in signal transduction. First, are the multiple signaling pathways activated by a single growth factor receptor used to activate gene expression, and if so, do these pathways act combinatorially? Second, how does the initial genetic response of a cell to a signal stimulus relate to the patterns of gene expression that determine that cell's ultimate biological response to the stimulus? Hill and Treisman take a critical look at what these array technology studies tell us concerning these questions and discuss technical issues arising from them. PMID- 11865183 TI - Getting directions: axon guidance receptors find the way. AB - As neuronal growth cones explore the terrain, their migration is dictated by guidance cues in the environment. Some cues are considered bifunctional because they can elicit an attractive or repulsive response. However, the cytoplasmic tails of guidance cue receptors appear to also control how growth cones respond. Merz and Culotti discuss recent evidence that the cytoplasmic domains of the DCC and UNC-5 family of receptors control the response of neurons to netrins, and propose a model for mechanisms by which axon guidance receptors might function. PMID- 11865184 TI - The NF-kappa B activation pathway: a paradigm in information transfer from membrane to nucleus. AB - Nuclear factor kappa B (NF-kappaB)/Rel proteins are dimeric, sequence-specific transcription factors involved in the activation of an exceptionally large number of genes in response to inflammation, viral and bacterial infections, and other stressful situations requiring rapid reprogramming of gene expression. In unstimulated cells, NF-kappaB is sequestered in an inactive form in the cytoplasm bound to inhibitory IkappaB proteins. Stimulation leads to the rapid phosphorylation, ubiquitinylation, and ultimately proteolytic degradation of IkappaB, which frees NF-kappaB to translocate to the nucleus and activate the transcription of its target genes. The multisubunit IkappaB kinase (IKK) responsible for the inducible phosphorylation of IkappaB appears to be the initial point of convergence for most stimuli that activate NF-kappaB. IKK contains two catalytic subunits, IKKalpha and IKKbeta, both of which phosphorylate IkappaB at sites phosphorylated in vivo. Gene knockout studies indicate that IKKbeta is primarily responsible for the activation of NF-kappaB in response to proinflammatory stimuli, whereas IKKalpha is essential for keratinocyte differentiation. The activity of IKK is regulated by phosphorylation. IKK contains a regulatory subunit, IKKgamma, which is critical for activation of IKK and is postulated to serve as a recognition site for upstream activators. When phosphorylated, the IKK recognition site on IkappaBalpha serves as a specific recognition site for the kappa-TrCP-like component of a Skp1-Cullin-F-box-type E3 ubiquitin-protein ligase. A variety of other signaling events, including phosphorylation of NF-kappaB, phosphorylation of IKK, new synthesis of IkappaBs, and the processing of NF-kappaB precursors provide mechanisms of modulating the amount and duration of NF-kappaB activity. PMID- 11865186 TI - All good things must come to an end: how is Notch signaling turned off? AB - Turning off signaling pathways can be just as important for proper biological regulation as turning them on. The Notch signaling pathway controls development of the nervous system in Drosophila. Proteolysis of Notch appears to initiate signaling, but further proteolysis may also terminate signals from this pathway. Kopan discusses mechanisms that limit signaling by Notch, including recent evidence that degradation of specifically targeted proteins by the proteasome is required. PMID- 11865187 TI - Signaling themes shared between peptide and steroid hormones at the plasma membrane. AB - Steroid hormone receptors are primarily thought of as transcriptional regulators that are activated by ligand binding. However, there is also evidence for "nongenomic" actions of steroid hormones. Cheryl Watson summarizes a recent meeting that brought together researchers exploring these rapid effects of steroid hormones that occur in the absence of new protein synthesis. Current evidence for the existence of membrane steroid receptors, which may or may not be related to the well-known nuclear receptors, is discussed. PMID- 11865188 TI - Tangled webs: evidence of cross-talk between c-Raf-1 and Akt. AB - The apparent cross-communication that can occur between different cell signaling pathways indicates that some signaling mechanisms may be more complex than originally envisaged. Jun et al. discuss recent studies suggesting that two signaling pathways that can be activated by the same growth factor receptor, the Ras-Raf pathway and the phosphatidylinositol 3-kinase (PI3K)--Akt (protein kinase B) pathway, can integrate with each other to generate a particular response, depending on the cell type and the stage of cell differentiation. PMID- 11865189 TI - EGF receptor transactivation mediated by the proteolytic production of EGF-like agonists. AB - The epidermal growth factor (EGF) receptor is activated not only by EGF-like ligands, but also by stimuli that do not directly act on the receptor, including agonists of G protein--coupled receptors and certain environmental stresses such as ionizing radiation. Carpenter discusses two reports that indicate EGF receptor activation by such heterologous stimuli may occur through the action of proteases that release cell surface EGF-like growth factor precursors. This mechanism of EGF receptor transactivation appears to involve the generation of soluble agonists. PMID- 11865191 TI - Manipulating Programmed Cell Death for Better Living! AB - Manolagas provides a succinct review of the book Apoptosis in Health and Disease. He praises the book for the clarity with which it covers the topics, but he also points out two missing health issues for which regulating apoptosis may prove clinically important: One is the health and regulation of the female reproductive system and the other is the treatment of osteoporosis. PMID- 11865192 TI - Thrombocytopenia in an animal model of malaria is associated with an increased caspase-mediated death of thrombocytes. AB - Infection of mice with Plasmodium Berghei Anka (PbA) leads to a thrombocytopenia, due to a reduced platelet life span, eventually associated with a syndrome of severe or cerebral malaria (CM). Thrombocytopenia was associated with an increase in the number of microparticles (mcp) in plasma. More than >60% of these mcp were of platelet origin, as seen by staining with an anti-platelet antibody. The thrombocytopenia and the amount of mcp were decreased in mice treated with anti CD40L mAb, suggesting that CD40L is the main effector of the thrombocytopenia. Caspase-1, -3, -6, -8, -9 were activated in platelets from infected mice, as seen by the binding of labeled probes or the amount of pro-caspase-3. Treatment of infected mice with the caspases inhibitor ZVAD-fmk decreased the number of mcp and the thrombocytopenia, showing that platelet caspases are responsible for platelet fragmentation. In addition, the caspase inhibitor also caused a decrease in the mortality associated with CM, indicating a critical role of caspases in the expression of CM. PMID- 11865193 TI - Effects of BAPTA-AM, Forskolin, DSF and Z.VAD.fmk on PDT-induced apoptosis and m THPC phototoxicity on B16 cells. AB - As many types of cells exposed to photodynamic therapy (PDT) appear to undergo apoptosis, various apoptosis inhibitors have already been used in studies of PDT induced apoptosis. Although these inhibitors decrease apoptosis, their real effect on the phototoxic efficacy of photosensitisers is unclear. The good phototoxicity of m-THPC was confirmed on murine melanoma B16-A45 cells. Toxicity and phototoxicity studies were then carried out using four apoptosis inhibitors: BAPTA-AM, Forskolin, DSF, and Z.VAD.fmk. Although all inhibitors tested blocked PDT-induced apoptosis, none produced a significant modification of the phototoxic effect of m-THPC on B16 cells. It has been suggested that apoptosis and necrosis share common initiation pathways and that the final outcome is determined by the presence of an active caspase. This implies that apoptosis inhibition reorients cells to necrosis, i.e. those cells sufficiently damaged by PDT appear to be killed, regardless of the mechanism involved. PMID- 11865195 TI - Calpeptin suppresses tumor necrosis factor-alpha-induced death and accumulation of p53 in L929 mouse sarcoma cells. AB - The cytokine tumor necrosis factor (TNF)alpha induces caspase-dependent cell death in a subset of tumor cells. In this report, we show a novel suppressive effect of calpeptin, a calpain inhibitor, on TNFalpha-induced cell death and accumulation of p53 in L929 mouse fibrosarcoma. Exposure to 10 ng/ml TNFalpha induced cell death in >50% of L929 cells within 12 h and stimulated accumulation of p53 (8-fold). Preincubation of cells with calpeptin blocked both TNFalpha induced cell death and accumulation of p53 as examined with Western blot. TNFalpha-induced accumulation of p53 was in part contributed by increase of p53 mRNA level (2.2-fold) in a calpeptin-insensitive manner. Interestingly, other calpain inhibitors tested did not show these effects like calpeptin and TNFalpha treatment did not increase apparent calpain activity in L929 cells, suggesting that calpeptin may have another function besides targeting calpain. Expression of dominant negative mutant p53Val(135) reduced the incidence of TNFalpha-mediated cell death. Taken together, our findings suggest that TNFalpha induces calpeptin dependent, but calpain-independent accumulation of p53 protein as a necessary step leading to death in L929 cells. PMID- 11865194 TI - Activation of caspase-8 is critical for sensitivity to cytotoxic anti-Fas antibody-induced apoptosis in human ovarian cancer cells. AB - Two ovarian cancer cell lines named NOS4 and SKOV-3 have been shown to have different sensitivities to a cytotoxic anti-Fas antibody, CH-11. Although both cell lines express Fas molecules on the cell surfaces at the same intensities, apoptosis is induced by CH-11 in NOS4 cells but not in SKOV-3 cells. In this study, the different apoptosis-sensitivities of these cells were assessed. Both cell lines express almost the same levels of FADD, RIP, c-FLIP, FAP-1, Bax, Bcl-2 and Bcl-XL. Evidence of caspase-8, caspase-9 and caspase-3 activation and of cleavage of PARP and Bid was obtained in NOS4 cells but not in SKOV-3 cells. When triggered by FasL protein, DNA fragmentation and caspase-8 activation were observed in SKOV-3 cells, though they were not as clear as in NOS4 cells. All the anti-Fas antibody-mediated signals for apoptosis induction in NOS4 cells were completely blocked by a caspase-8-specific inhibitor, Z-IETD-FMK. These results indicate that the different sensitivities to the anti-Fas antibody are solely dependent on the activation of caspase-8, which could be influenced by yet unknown qualitative or quantitative abnormalities in molecules involved in DISC formation. PMID- 11865196 TI - Commitment to apoptosis induced by tumour necrosis factor-alpha is dependent on caspase activity. AB - Tumour Necrosis Factor alpha binding at the cell surface induces a complex series of signaling events culminating in the caspase cascade, which is central to apoptosis. However, recent work from several laboratories has questioned caspase involvement in commitment to cell death. We have therefore investigated the involvement of caspases in the crucial commitment stage of tumour necrosis factor induced apoptosis in human T-leukaemic CEM-C7 cells and breast carcinoma MCF-7 cells, using both peptide-based and viral caspase inhibitors. Our observations converge on the conclusion that commitment to death in these systems is dependent on caspase activity, e.g. baculovirus p35 produces over 50-fold protection of colony-forming ability, the most stringent criterion of cell survival. These observations strongly support the view that the caspase family is of great biological and medical significance, since caspase dysfunction resulting in failure to commit to cell death after treatment with tumour necrosis factor or other stimuli may contribute to cancer development. PMID- 11865197 TI - Apoptosis and apoptosis related proteins in chronic viral liver disease. AB - BACKGROUND: Apoptosis may be an important mechanism of hepatocyte death in chronic viral liver disease. METHODS: We studied apoptosis in liver biopsies from 30 patients with chronic viral hepatitis and 8 patients with viral cirrhosis by the TUNEL method. 12 cases of non-alcoholic steatohepatitis and 12 cases of primary biliary cirrhosis were used as non-viral disease controls. Immunohistochemical expression of p53, p21/waf1, bcl-2 and mdm-2 proteins was also studied in the same patients. RESULTS: A statistically significant increase of apoptotic liver cells was found in severe chronic viral hepatitis (5.3 +/- 0.3%), cirrhosis (3.4 +/- 0.5%) and PBC (4.4 +/- 0.4%) cases compared to patients with non-alcoholic steatohepatitis (0.8 +/- 0.3%). The expression of p53 protein was increased in the cases of viral cirrhosis and in chronic severe viral hepatitis whereas in the cases of chronic mild hepatitis, PBC and non-alcoholic steatohepatitis we found no expression of p53. P21/waf1 expression was increased in severe chronic hepatitis, cirrhosis and PBC cases compared to mild hepatitis and non-alcoholic steatohepatitis cases. However no induction of mdm-2 was observed in the subgroups of chronic liver disease. Bcl-2 was expressed only in epithelium of bile ducts and mononuclear cells of the portal tracts and liver lobules. A weaker Bcl-2 expression was noted in the epithelium of bile ducts of 7/12 PBC cases. CONCLUSION: Our results provide evidence of increased apoptosis in severe chronic viral liver disease, suggesting that apoptotic cell death might be involved in the pathogenesis of hepatocellular damage of viral hepatitis and cirrhosis. Furthermore we analysed part of the apoptotic pathways implicated in the above process and found an increased expression of p21/waf1, probably p53 mediated, without overexpression of the apoptosis inhibiting bcl-2 and mdm-2 proteins. By contrast p21/waf1 overexpression in PBC seems to be propagated by a p53 independent mechanism. PMID- 11865198 TI - Actin involvement in apoptotic chromatin changes of hemopoietic cells undergoing hyperthermia. AB - During apoptosis, cell chromatin undergoes characteristic morphological changes, which have been long described in a variety of experimental models but are mostly not yet understood. The aim of the present study was to investigate the mechanisms underlying this phenomenon and the possible role of cytoskeleton, in particular actin. The chosen apoptotic model were HL60 hemopoietic cells undergoing hyperthermia and the starting point was the observation of thin filament bundles in decondensed chromatin of their early apoptotic nuclei. The characterization of these structures was undertaken by cytochemical, fluorescent and immunogold techniques, directed to actin identification. Taken together, our results suggest, in apoptotic cells, a deep actin rearrangement. Moreover, this cytoskeletal component, never present in normal nucleus, appears in the early apoptotic one, where it can be found in polymerized form, promptly recognizable both by conventional and immunogold electron microscopy. We suggest that, similarly to the role played by nuclear matrix in interphase and mitotic nucleus, actin could be directly involved in chromatin rearrangement occurring in apoptosis. PMID- 11865199 TI - The disposal of dying cells in living tissues. AB - Cells continuously die and disappear from the midst of living tissues. However, some of their constituents survive. DNA is horizontally transferred to phagocytic cells, and apoptotic cell antigens shape the immune repertoire. When massive apoptosis occurs, which overwhelms tissue scavenger cells, or when the function of phagocytes abates, dying cells escape clearance in vivo. Remnant dying cells come to phagocytes disguised: factors capable to envelop their membranes pervade the entire organism, or are generated in given tissues. Some are constitutively present, while other are generated during early or late phases of the inflammatory response, possibly to face the further burden of the dead inflammatory cells. This camouflage influences the disposal of the corpses: decoying molecules either bridge the corpse to the phagocyte or hide it. Furthermore, factors associated to the plasma membrane of the apoptotic cell shape the signals the phagocyte releases in situ. Finally, molecules contained or released by the dying cell alter the apprehension by the phagocyte of its prey, influencing its immunogenicity. PMID- 11865200 TI - Cell death regulation by the mammalian IAP antagonist Diablo/Smac. AB - In Drosophila, the genetic locus 75CI1,2 is essential for all developmental cell death. Within this region are the genes for three pro-death proteins, Grim, Reaper and HID. These proteins are transcriptionally regulated and their expression tightly associated with cell death in the developing fly embryo. When ectopically expressed in the retina, Grim, Reaper and HID cause apoptosis and eye ablation. They have a short region of similarity at their N-termini through which they can interact with inhibitor of apoptosis (IAP) proteins, and it is by antagonising IAP inhibition of caspases that Grim, Reaper and HID promote cell death. The observation that Grim, Reaper and HID can interact with mammalian IAPs and induce apoptosis in mammalian cells suggested that mammalian IAP antagonists might also exist. Diablo/Smac, identified six years after the first description of a Drosophila IAP antagonist, is the only mammalian protein identified to date that is clearly functionally related to the Drosophila proteins. Since its discovery, there have been numerous studies investigating how Diablo/Smac interacts with IAPs and promotes cell death. Here we review what is currently known about Diablo/Smac and speculate on other mammalian IAP antagonists. PMID- 11865201 TI - The regulation of APAF1 expression during development and tumourigenesis. AB - Apoptosis Protease-Activating Factor 1, APAF1, was originally isolated four years ago and shown to be the mammalian homologue of the C. elegans pro-apoptotic ced4 gene. Since then, the expression of APAF1 has been demonstrated to be involved in several cell death pathways, including the induction of apoptosis by the p53 tumour suppressor protein and neuronal apoptosis. In this review we will focus on the regulation of APAF1 expression, in particular with regard to recent developments in our understanding of the role of APAF1 in both tumourigenesis and mammalian development. PMID- 11865202 TI - Surviving apoptosis. AB - The concept that cells subjected to chromatin cleavage during apoptosis are destined to die is being challenged. The execution phase of apoptosis is characterized by the activation of effector caspases, such as caspase-3, that cleave key regulatory or structural proteins and in particular activate apoptotic nucleases such as the caspase activated deoxyribonuclease (CAD). It is apparent that caspases of this type may become active both through non-apoptotic processing and potentially within cells that exhibit apoptotic morphology but are subsequently able to survive. In such systems caspase suppressor molecules, the inhibitors of apoptotic proteins or IAP's, may rescue cells from apoptotic nuclease(s) attack initiated by transient caspase activation. The MLL gene is involved in leukemogenic translocations in ALL and AML and is a target of nuclease cleavage during apoptosis. Translocations initiated at the site of apoptotic nuclease attack within MLL have been identified and may offer a model, with clinical relevance, for DNA damage mediated by the apoptosis system in cells destined to survive. The specificity of apoptotic cleavage combined with the potential for recovery from the execution phase of apoptosis suggests a novel and pathogenic role for apoptosis in creating translocations with leukemogenic potential. PMID- 11865203 TI - Vitamin E analogs: a new class of multiple action agents with anti-neoplastic and anti-atherogenic activity. AB - The incidence of cancer and atherosclerosis, two most common causes of death in developed countries, has been stagnating or, even, increasing. Drugs effective against such conditions are needed and, in this regard, the potential anti atherosclerotic activity of vitamin E analogs has been studied extensively. Surprisingly, recent results indicate that these agents may also exert anti neoplastic effects. Here we review the evidence that particular analogs of vitamin E may act as both antiatherogenic and anti-cancer agents, and discuss the possible molecular bases for these actions. PMID- 11865204 TI - Walk-in centres: not yet on a firm footing. PMID- 11865205 TI - Clean intermittent self-catheterisation and children. AB - The best chance for good renal health and continence for children with a neuropathic bladder is clean intermittent self-catheterisation. In order to maximize potential for good renal health and continence the child and family need to be well supported by the multidisciplinary team, making good use of the knowledge of the children's urology specialist nurse who is able to provide relevant information, advice and education. With support many children with a neuropathic bladder are able to achieve continence while maintaining good renal health. PMID- 11865206 TI - Providing seamless community health and social services. AB - This article reviews a variety of Government community health policies and initiatives, from the NHS and Community Care Act 1990 to the emergence of primary care groups (PGCs). The Labour administration abolished GP fundholding and has placed considerable emphasis on its commitment to partnership with the care professions. The Government is also expecting a commitment to partnership between the community care professions in order to reflect the entwined social and health care needs of the individual requiring care in the community. The role of the district nursing service has been brought into focus with the opportunity of representation at PCG board level. Evaluating how a district nursing service may integrate with social services is therefore, and appropriate strategy for assessing whether recent policy initiatives go far enough at grass roots level in facilitating collaboration and quality community care. PMID- 11865207 TI - Managing gravitational eczema and allergic contact dermatitis. AB - Venous leg ulceration is a common chronic problem in the community, and gravitational eczema is a common complication of it. The presence of gravitational eczema can lead to further ulceration and may also impair wound healing. Treatment often requires more than one topical preparation, to gain control and prevent relapse. Knowledge of the benefits and risks of these therapeutic modalities is essential, in this article we aim to provide a practical approach to the treatment of gravitational eczema. We also discuss allergic contact dermatitis, a common complication of gravitational eczema. PMID- 11865208 TI - Common stoma problems: a brief guide for community nurses. AB - It is estimated that there are 100,000 people in the United Kingdom who have a stoma, some 47% of whom will suffer at least one complication (Lyon and Smith, 2001). District nurses work with specialist stoma care nurses to manage the majority of these complications, so it is vital that they maintain their knowledge. This article reviews some of the stoma complications that many arise, and uses two case studies to illustrate how these may be dealt with in the community. PMID- 11865209 TI - Threadworms: a starting point for family hygiene. AB - Owing to the embarrassing anal itch caused by threadworm infection, some sufferers feel they cannot even mention the problem to their doctor. Community nurses, often regarded as a more approachable source of support, will also be able to prescribe medication if they have prescribing status. With an adequate understanding of the complaint, they can offer a choice of treatment options, mechanical removal or drugs, necessarily backed by the appropriate personal and environmental hygiene measures. The starting point for relevant hygiene is cheerful good practice in hand washing at home and school, which is also key to preventing the spread of many other more serious infections. Implementation is of basic importance and should form part of any of the current initiatives from Sure Start to Healthy Schools. PMID- 11865210 TI - OpSite Plus in the management of exuding wounds. AB - This article discusses the use of OpSite Plus (formerly known as OpSite Post-Op ) in the community setting. This semi permeable film dressing with an absorbent pad was evaluated by the community hospital and by practice and district nurses within Plymouth's NHS Primary Care Trust. PMID- 11865211 TI - No place for prejudice or discrimination. PMID- 11865212 TI - Politics is undermining health care. PMID- 11865213 TI - Developing wound management guidelines for community nurses. AB - Quality improvements are at the forefront of the Government's agenda and the advent of clinical governance has increased interest in the evidence base on which clinical decisions are made. Clinical guidelines are seen as an important tool in achieving evidence-based practice and there has recently been a proliferation in guideline development, nationally and locally. This article describes the rationale for the development of wound management guidelines for community nurses and the key steps that were involved in this process. It discusses and highlights the importance of an implementation strategy, and the need to evaluate, review and ensure that guidelines are up to date. It is intended to be of use to those nurses in the community who are considering undertaking guideline development. PMID- 11865214 TI - Flexible cystoscopy: outpatients or domiciliary? AB - The cost of caring for people in the community with long-term indwelling catheters varies widely but represents a significant amount of NHS spending. A rapid access catheter clinic was established at a Bristol hospital, for people experiencing difficulties with their long-term catheters, to facilitate prompt assessment and treatment. Many patients were found to be severely disabled with mobility problems and relied on hospital transport to attend the clinic for flexible cystoscopy. As a consequence the concept of undertaking this procedure on a domiciliary basis was introduced with a view to cutting costs, improving time management and offering a more reliable service to patients. A prototype battery-powered flexible cystoscope was developed to facilitate this service and five visits were undertaken. Feedback from patients indicated a preference for the domiciliary service and cost benefits were identified. As a result of these initial experiences, we are now planning a pilot study to establish the feasibility and costs of providing a domiciliary service on a permanent basis and gather qualitative data from patients on quality-of-life issues. PMID- 11865215 TI - A survey of drinking and toilet facilities in local state schools. AB - The public health role of the school nurse has been formalized recently in several documents. Within that role, they are charged with assessing and responding to health needs of the school-aged population. Children and young people are often accused of making unhealthy choices with regard to their fluid intake, however, there have been informal reports that it is difficult to access water in school. Also, with inclusion for all children high on the agenda, do schools have adequate toileting facilities for children with special needs? This survey used school nurses to assess the access to Midlands schools. It also assessed the cleanliness and facilities available for hand washing and other sanitary processes. It found that many schools are failing to provide the facilities they are expected to. Overcrowding and potentially insanitary conditions were found in a significant proportion of the schools surveyed. Drinking facilities and access was found to be poor. Recommendations are made to begin to address the problems seen here. PMID- 11865216 TI - Do poor nutrition and display screens affect visual acuity in children? AB - Two studies conducted in Scotland have shown an increase in visual acuity (VA) screening failure among primary school-aged children in recent years. Two other trends were observed during the same period - an increase in children bringing packed lunches to school, and increased access to display screen equipment (DSE) including television, computers and hand-held computer games. This study set out to assess if either DSE use of poor diet could be linked with visual acuity screening failure in Scottish primary school children. Information was collected on diet and DSE use from 1384 children who had received VA screening in eight primary schools in Glasgow. After controlling for deprivation, DSE use other than watching television was associated with an increased risk of VA screening failure, as was a 'poor' diet. If these findings are confirmed by other studies and a causal link can be made, then health education at school could be designed to prevent VA screening failure. PMID- 11865217 TI - Maintaining and developing competencies in nurse prescribing. AB - The development of nurse prescribing in England has taken a step forward with the introduction of the new extended curriculum in January 2002. However, the extension will refocus attention on the safety and quality of the care delivered by nurses who prescribe. Individual nurses have a responsibility to practice safely, while their employing organizations have a responsibility to put in place the systems to ensure this. The recently published document 'Maintaining Competency in Prescribing: An Outline Framework to Help Nurse Prescribers' (National Prescribing Centre, 2001) aims to help both parties achieve this. PMID- 11865218 TI - The most effective products available to facilitate ear syringing. AB - Loss of hearing from accumulation of cerumen (ear wax) is a commonly seen problem. Treatment for this condition often involves use of a wax softening or dispersing agent (cerumenolytic) before syringing. A mini-review was undertaken to find out what is the best preparation to use before syringing to facilitate the procedure. Five databases were searched using subject searches combined with methodological filters to find the highest level of evidence to answer the question. Nine randomized controlled trials (RCTs) were found and seven in vitro studies that researched the efficacy or effectiveness of a range of products. The RCTs do not show any one product to be more effective. The in vitro studies show Waxsol to be better at dispersing wax followed by sodium bicarbonate. This needs to be demonstrated with in vivo testing comparing all products for effectiveness. Many of the studies suffered from a lack of information. Side effects were not adequately investigated. PMID- 11865219 TI - Complementary therapies and health promotion. AB - Nurses have an important role to play in health promotions and it is important that they have a comprehensive knowledge and understanding of the theories of health promotion and conventional interventions. However, they are increasingly likely to be asked about the efficacy of non-conventional interventions and the likelihood of these interventions being of value to the client. This paper offers a review of the evidence to support the role of complementary therapies as adjunctive treatments to enable nurses to discuss their use with their clients as part of their health choice options. PMID- 11865220 TI - The politics of nurse prescribing. PMID- 11865221 TI - Bringing the outsiders in: tackling social exclusion. PMID- 11865222 TI - Principles for pressure ulcer prevention in the community. AB - This article aims to examine the prevention of pressure ulcers in the community. Pressure ulcers continue to cause needless distress and anxiety for patients, carers and their families. Health professionals in the community often face extreme challenges when caring for patients who are at risk of developing pressure ulcers. This could be due to lack of appropriate assessment skills and having insufficient time to conduct an appropriate assessment. Having an understanding and awareness of the function of the skin and the aetiology of pressure ulcer development can assist the health professional in attempting to take a proactive approach to prevent pressure ulcer formation in the community setting. PMID- 11865223 TI - An assessment of dual-role primary care nurses in the inner city. AB - The integration of primary care nursing roles into single posts, closely associated with GPs, has been advocated in the UK and in Europe. However, there has been little exploration of the benefits and difficulties of integrating roles in the UK. In this article, we assess three posts in inner London that combine health visiting and district nursing. Our findings indicate potential benefits in collaborative working with small general practices, as well as high job satisfaction in post-holders. However, these benefits, when weighed against consequences such as increased stress for the post-holders in managing two separate caseloads and increased coasts for the nursing services, would need to be assessed in a larger-scale study. Our study suggests that other combinations of current, defined specialties in community nursing might capitalize on the benefits and diminish the negative consequences. PMID- 11865224 TI - The patient's view: the benefits and limitations of nurse prescribing. AB - This study was undertaken in a primary care group to explore nurse prescribing from the patient/client's viewpoint. All prescribing health visitors, district nurses and practice nurses were asked to recruit five patients for whom they had prescribed; 50 patients/clients participated in the study. Identified benefits of nurse prescribing included a more effective use of the nurse's and doctor's time; a quality relationship between the nurse and patient; nurses' awareness of their own professional limitations; their expertise in certain types of care; and their providing timely, convenient, practical and successful treatment. Limitations and the proposed options for change included the training and competency of nurse prescribers and the limitations of the Nurse Prescribers' Formulary. On a local level the study informs nurse prescribers that they are currently meeting the needs of the majority of recipients, and provides evidence of some of the benefits and limitations of nurse prescribing. PMID- 11865225 TI - Nurse prescribing: views on autonomy and independence. AB - The introduction of nurse prescribing throughout Scotland in the primary care setting has proved to be an interesting development for nurses. The aim of this study was to assess the impact of nurse prescribing on district nurses and health visitors in one NHS trust in Scotland. All prescribing nurses in the trust were asked to participate in the study. This explored their perceptions of autonomy and the level of dependence on GP colleagues since being able to prescribe. In addition, prescribing patterns were examined to ascertain whether there was a relationship between the area of nursing (e.g. district nursing or health visiting) or the length of prescribing experiences, and the number of prescriptions written. PMID- 11865226 TI - Compression hosiery in venous insufficiency: a natural nurse-led field. AB - The use of compression hosiery for the treatment and prevention of venous ulceration caused by venous insufficiency is commonplace in community practice. In recent years the role of the nurse in the management of these patients has grown supported by increasing levels of knowledge, nurse specialists and the implementation of nurse prescribing. The full list of compression hosiery items available on NHS prescription are included in the list of nurse prescribing items in the Nurse Prescribers' Formulary and the Drug Tariff. Central to a successful treatment outcome is the patient assessment. This forms the base of a partnership approach to care upon which product choices and advice can be tailored to meet the individual needs of that patient. An understanding of venous physiology, limb measurement skills and hosiery care place the nurse in an ideal position to lead in this area of nursing practice. PMID- 11865227 TI - The role of alternative medicine in treating postnatal depression. AB - Postnatal depression is a serious and debilitating condition. Due to the perceived stigma of mental illness, the incidence of it is under-reported and many mothers refuse psychiatric help, either assuming postnatal depression to be normal or because of the potential consequences of having a psychiatric history. Community practitioners who are in contact enhance the supportive care they give to these women. This article discusses the evidence for a number of these interventions which mothers may find more acceptable than orthodox treatment. PMID- 11865228 TI - Awful things can happen inside the home. PMID- 11865229 TI - The way in which we address patients can be undermining. PMID- 11865230 TI - Government response to Bristol inquiry: is it too ambitious? PMID- 11865231 TI - Nurse who tied down elderly patients and used cot slides. PMID- 11865232 TI - A review of amputation, phantom pain and nursing responsibilities. AB - Phantom pain is a common phenomenon among amputee patients. It is usually reported during the immediate postoperative period but can persist for up to 2 years. Sixty per cent of patients can still experience phantom pain 7 years postamputation and it has even been reported 30 years postamputation. Pain of any type, which persists for more than 7 months, is difficult to treat and is often unresponsive to analgesia, including opioids. A large percentage of patients undergoing amputation have experienced chronic pain before their operation. Chronic pain is known to affect adversely patients' quality of life, and ability to function and achieve quality sleep. Nurses, therefore, need to address not only the physical aspects of amputation and phantom pain but also the psychological aspects. PMID- 11865233 TI - Nurses' education and skills in bandaging the lower limb. AB - Given that community nurses are the primary professional carers of patients presenting with leg ulcers, this article considers and argues for nurses to be educated once more in the precise skills of bandaging the lower limb. Although in the early part of the 20th century bandaging techniques were precisely taught as part of basic nurse education, modern nursing texts and curriculum fail to give the subject the attention it deserves to ensure patients receive high-quality care. PMID- 11865234 TI - Self-disclosure in adult and mental health nursing students. AB - Self-disclosure, the process whereby people verbally reveal themselves to others, is an important interpersonal skill in initiating, developing, maintaining, and terminating therapeutic relationships. Studies of this interpersonal skill in nursing have, primarily, focused on adult nurses. They are dated, make no comparisons between this and other nursing specialisms, and have not considered the patient as a focus for nurses' self-disclosure. This study attempts to address these issues in relation to student nurses by: replicating a previous study of self-disclosure in adult nursing students (ANS) and offering a comparison of two nursing specialisms, reporting on the use of this skill in relation to the patient. A matched sample of 25 ANS and 25 mental health nursing students (MHNS) were asked to complete a modified version of Jourard's 25-item self-disclosure questionnaire. The findings suggest that: ANS were more self disclosing than students sampled in an earlier study (Burnard and Morrison, 1992); both sets of students disclosed significantly less items to the patient as target-person than to other categories; and MHNS disclosed significantly more items than the ANS to the patient category. PMID- 11865235 TI - A survey of 'teamwork' in mental health: is it achievable in practice? AB - As part of an ENB educational evaluation of teamworking in mental health, a report-and-respond survey was used as one of five data collection methods. This instrument disseminated earlier findings back to the participants and invited further comments. This article discusses the usefulness of this type of survey as a data collection tool and presents the findings of the survey which was administered to service professionals, educationalist, students, users and carers involved in mental health throughout England. The findings show that when groups of professionals are working within increasingly tight financial constraints and increased workloads, multiprofessional teamworking can be scarce in reality. These tensions present issues for nurse education and training at both pre- and post-registration levels in relation to the teaching of, for and about effective teamwork. PMID- 11865236 TI - Pulse oximetry: an audit of nursing and medical staff understanding. AB - Pulse oximetry provides continuous, non-invasive monitoring of the oxygen saturation of haemoglobin in arterial blood, which is updated with each pulse wave. The primary advantage is that it detects immediately deviations to a patient's baseline. Care givers can therefore detect changes before the patient becomes hypoxaemic. However, staff are reported to have limited education, if any, in the correct use of the pulse oximeter and what may affect the readings. In order to test this theory, an audit was conducted on 50 staff within a large general hospital, comprising trained and untrained nurses and medical staff. Participants' responses to a questionnaire and to six clinical scenarios were analysed. Overall, there was a deficit in participants' knowledge on pulse oximetry. The answers given by the medical staff to the clinical scenarios showed no greater level of knowledge than many of the trained nurses. The recommendations made following this audit were to improve the education and training of all staff in the understanding of this technique. PMID- 11865237 TI - Legal aspects of consent 21: amputations of healthy limbs. PMID- 11865238 TI - Avance: silver hydropolymer dressing for critically colonized wounds. AB - Avance is a hydropolymer dressing that has a silver compound bonded into it. The silver acts as a bacterial shield to prevent bacterial invasion, and as a bacterial barrier to impede cross-infection. This article aims to give an overview of the use of silver in eradicating surface bacteria, and provides case study evidence of the use of Avance on the leg ulcers of two patients. Both patients had complex medical histories and underlying aetiologies that delayed the wound-healing process. PMID- 11865240 TI - Nursing needs to start proving its value. PMID- 11865239 TI - Anticipating the patient's needs: a key nursing skill. PMID- 11865241 TI - Record keeping should be a key skill not a chore. PMID- 11865242 TI - Nurse who lacked the skill to care for a patient with diabetes. PMID- 11865243 TI - Psychological wellbeing of patients with rheumatoid arthritis. AB - The impetus for this multiphased project was the realization that rheumatoid arthritis (RA) is not only physically debilitating, but also a cause of severe psychological distress which is often unacknowledged. A small sample of patients (n=15) was interviewed to identify their main area of psychological distress caused by RA, and data were analysed thematically. These themes formed the basis of a questionnaire that was distributed to a random sample of inpatients and outpatients with RA. This larger sample (n=140) prioritised the areas of distress that were identified by the interviewed sample of patients. This led to the development of an education programme for nurses. The authors describe the impetus for the project, the activities and processes associated with data collection, and the development and evaluation of the education programme. PMID- 11865244 TI - Legal aspects of consent 17: not for resuscitation instruction. PMID- 11865245 TI - The pharmaceutical industry and mental health nursing. AB - Mental health nurses (MHNs) have a long and varied relationship with the pharmaceutical industry, including the attendance of educational seminars and the acceptance of promotional material. This article presents evidence that suggests MHNs may need to review this relationship regardless of their beliefs about the role of pharmacological interventions in the treatment of mental illness. It also explores the influence of advertising within professional journals and calls for greater debate on this issue. PMID- 11865246 TI - The dynamics and processes of 'ending' in clinical supervision. AB - While there is a growing number of papers in academic and professional nursing journals that focus on clinical supervision, there remain unanswered questions and unresolved issues. One such issue where there is a distinct paucity of theoretical or empirical work is that of 'ending' within clinical supervision. Accordingly, this article examines key issues and dynamics of ending within clinical supervision. These key issues are summarized as: ending when the supervisor is reluctant to let go; parallel processes in ending in clinical supervision and ending in clinical relationships; the different ending dynamics of different approaches to clinical supervision; endings in group supervision; ending as a form of bereavement; healthy endings in clinical supervision; and endings as a opportunity for growth and celebration. The authors posit that an understanding of these processes can help facilitate a 'healthy' ending in clinical supervision. Furthermore, there may be particular merit in considering dynamics that, when present, create the best chance of all parties experiencing a health ending, namely: the ending is negotiated; the ending is gradual rather than sudden and all parties work towards the ending; the supervisee retains (wherever possible) a degree of control over the timing of the ending; and both supervisor and supervisee achieve a sense of closure. PMID- 11865247 TI - The biosciences and fitness for practice: a time for review? AB - The learning of biosciences by preregistration students has been recognized as being problematic, and the need for better resourcing of education has been identified. The Project 2000 initiative (UKCC, 1988) expanded the breadth and dept of the curriculum content in order to support the delivery of holistic care, and so the resources for bioscience education seemed likely to remain limited. However, the UKCC has concluded that there has been a shortfall in the acquisition of practice skills by preregistration students (UKCC, 2001). To address this, new directives indicate that curricula should move to competency based outcomes and use student-focused learning. However, there is a lack of clarity as to what the basic expectation is for bioscience learning at registration, and how this might be continued in postregistration programmes. The authors argue that if resources for bioscience teaching in preregistration curricula cannot be improved then it is time to review the situation to ensure that expectations are realistic and attainable. Criteria are required to establish exactly what life-long learning means in relation to the biosciences, from registration through to specialist practice. PMID- 11865248 TI - Overview of research to investigate pressure-relieving surfaces. AB - Pressure-relieving equipment plays a key role in the prevention and treatment of pressure ulcers. Every year an increasing amount of equipment is launched on to the market. The efficacy of this equipment is traditionally qualified with interface pressure and case studies, but rarely with randomized controlled trials. With the advent of The New NHS: Modern, Dependable (Department of Health (DoH), 1997) and Pressure Ulcer Risk Assessment and Prevention (National Institute for Clinical Excellence (NICE), 2001), a clinical governance system has started to be put into place and hopefully more evidence will be produced by the manufacturers and the NHS. With some 200 different types of mattresses on the market, emphasis needs to be placed on good quality randomized clinical trials to establish effectiveness of what can be costly equipment. This article gives an overview of how, historically, pressure-relieving mattresses were marketed on case studies and interface measurement. Recently, randomized controlled clinical trials are being used to demonstrate the efficacy of the mattress and reduction in the incidence of pressure ulcers. Seating is also an important aspect in continuing 24-hours pressure area prevention and treatment. Patient posture in a chair needs to be fully understood by the practitioner and key strategies are made in this article. Little research into seating has been undertaken and is urgently required. PMID- 11865249 TI - Introducing the pressure support surfaces from Kaymed. AB - The range of support surfaces available is quite varied. It is important that both the purchaser and the users are satisfied with the quality, comfort and the pressure-reducing properties of the mattress. The mattress needs to be versatile, in that it can be used both in a patient's home or in a hospital or nursing home and also on profiling beds. The Kaymed range of mattresses offers increased comfort with low interface pressures for patients up to and including high risk. This product focus examines the Kaymed mattress and looks at the design and results of tests performed on the visco-elastic foam used in the manufacturing of the mattresses. PMID- 11865250 TI - How relevant to nursing is your degree? PMID- 11865252 TI - Undue influence and elder abuse: recognition and intervention strategies. AB - Undue influence is the substitution of one person's will for the true desires of another. Unlike common persuasion and sales techniques, such influence often entails fraud, duress, threats, or other deceits and pressures. Undue influence takes place when one person uses his or her role and power to exploit the trust, dependency, or fear of another to gain psychologic control over the weaker person's decision-making, usually for financial gain. Dependent and impaired people are particularly susceptible, but it can happen to anyone who otherwise would be considered capable and competent. The current interest in undue influence represents the union of three major forces: current and historical legal concepts, knowledge learned from domestic violence, specifically the field of elder abuse and neglect, and the distillation of relevant psychologic processes. PMID- 11865253 TI - Testing an intervention to reduce assaults on nursing assistants in nursing homes: a pilot study. AB - Nursing home staffs frequently encounter physical and emotional violence directed toward them by residents. This article addresses the problem from an occupational health perspective and describes a pilot study focused an educational intervention to reduce resident assaults on certified nursing assistants (CNAs) in nursing homes. After a 4-hour education intervention, the study CNAs reported fewer physical assaults by residents and increased levels of knowledge and confidence in their ability to manage residents' aggressive behavior compared with a similar CNA group. Limitations include a small convenience sample who works only the day shift. Results support education and training as effective in reducing resident physical assaults and enhancing nursing home safety. Further research should investigate assaultive resident behavior and evaluate the effectiveness of interventions to prevent, reduce, and manage violence in nursing homes. PMID- 11865254 TI - Initiating a long-term care nursing service for aging inmates. AB - In establishing a new long-term care nursing service in a correctional health care setting, the director of nursing (DON) sets the organizational milieu with a holistic humanistic perspective. She is expected to demonstrate the leadership attributes of vision, creativity, and accountability as manifested through outcomes. Based on her 6-year experience, the author delineates the DON's role in collaboratively initiating a program at a new facility, devising a method to identify geriatric inmates in need, and establishing a hospice program in a federal medical center where security takes precedence. PMID- 11865255 TI - Health promotion for aging adults. AB - This article describes the essentials of health promotion, aging in America, and factors to consider when developing health promotion materials for elders. It also provides a checklist to evaluate materials for appropriateness. PMID- 11865256 TI - Overview of the nursing home litigation process. AB - An increasing number of nursing home litigations have been filed as a result of the growing nursing home population and laws regulating their care. Understanding the litigation process and developing an awareness of the issues examined by attorneys and experts for both the plaintiff and defense are important for nurses working in long-term care. Knowing the standards of care and the litigation process can prepare nurses to anticipate and successfully defend their positions. PMID- 11865257 TI - Rita K. Chow, EdD, RNC, HNC, FAAN: a life in the light. PMID- 11865258 TI - Assessing the gifts, talents, and skills of nursing home residents. PMID- 11865259 TI - Recently approved and forthcoming drugs for elders. PMID- 11865260 TI - Social isolation: important construct in community health. PMID- 11865261 TI - Career development for physician-scientists: the model of the Pediatric Scientist Development Program. PMID- 11865262 TI - Cord serum ferritin levels, fetal iron status, and neurodevelopmental outcomes: correlations and confounding variables. PMID- 11865263 TI - The impact of molecular genetics on the clinical management of pediatric sensorineural hearing loss. PMID- 11865264 TI - Current status of radiofrequency ablation for common pediatric supraventricular tachycardias. PMID- 11865265 TI - Cystic fibrosis: a 2002 update. PMID- 11865266 TI - Cord serum ferritin concentrations and mental and psychomotor development of children at five years of age. AB - OBJECTIVE: Our purpose was to evaluate the association between fetal iron status and mental and psychomotor development at 5 years of age. STUDY DESIGN: We evaluated the association of fetal iron status (umbilical cord serum ferritin concentrations) with test scores of mental and psychomotor development of 278 children. Six tests were given, including full-scale intelligence quotient (FSIQ), language ability, fine- and gross-motor skills, attention, and tractability. RESULTS: Compared with children with cord ferritin in the 2 median quartiles, those in the lowest quartile scored lower on every test and had significantly worse language ability, fine-motor skills, and tractability. They were also 4.8-fold more likely to score poorly in fine-motor skills and 2.7-fold more likely to have poor tractability than children in the median quartiles. FSIQ in the highest quartile was slightly, but not significantly, lower than the median quartiles, but the odds ratio for having a FSIQ score of less than 70 for children in the highest quartile was 3.3 (95% CI 1.2-9.1). CONCLUSION: Poor iron status (low ferritin) in utero appears to be associated with diminished performance in certain mental and psychomotor tests. The reason for the association between high ferritin concentrations and low FSIQ scores is unknown. PMID- 11865267 TI - Chorioamnionitis, mechanical ventilation, and postnatal sepsis as modulators of chronic lung disease in preterm infants. AB - OBJECTIVE: This case-control study of chronic lung disease (CLD) evaluated the hypothesis that chorioamnionitis promotes CLD and interacts with other risk factors for CLD, including mechanical ventilation and postnatal infection. STUDY DESIGN: We identified a population of 193 infants who met our case criteria for CLD whose birth weights were 7 days and culture documented sepsis. In multivariable analyses, infants were at greatest risk for CLD when they had exposure to both chorioamnionitis and either mechanical ventilation >7 days (odds ratio, 3.2; 95% confidence interval, 0.9-11) or postnatal infection (odds ratio, 2.9; 95% confidence interval, 1.1-7.4). CONCLUSIONS: We conclude that prolonged mechanical ventilation or postnatal infection increases the risk of CLD among surviving preterm infants and that these 2 factors interact with antenatal infection to further increase the risk of CLD. PMID- 11865268 TI - Acute respiratory failure and short-term outcome after premature rupture of the membranes and oligohydramnios before 20 weeks of gestation. AB - OBJECTIVES: To study the effect of premature rupture of membranes and oligohydramnios before 20 weeks of gestation (PROM20) on acute respiratory morbidity and on short-term outcome in infants with a gestational age >or=24 weeks. STUDY DESIGN: A historic cohort study was performed of all infants born after PROM20 with a gestational age greater-than-or-equal24 weeks between 1990 and 1999. Control infants were matched for year of birth, gestational age, and birth weight. RESULTS: PROM20 infants had an increased acute respiratory morbidity (higher ventilator settings and increased incidence of hypoxemia, hypercapnia, and pulmonary hypertension) and a trend to more air leaks. Although not statistically different, PROM20 infants had more complications (neonatal survival, 68% vs 95%; severe intracranial hemorrhage, 31% vs 6%; chronic lung disease in surviving infants, 46% vs 17%). The relative risk for combined morbidity (death, intracranial hemorrhage, chronic lung disease) was increased (3.0, P =.019) when compared with matched control infants. However, 31% of the surviving PROM20 infants were discharged without apparent morbidity. CONCLUSIONS: Expectant treatment in women with PROM20 and present neonatal intensive care has improved the survival of PROM20 infants despite severe initial respiratory failure. However, chronic morbidity still occurred. PMID- 11865269 TI - Randomized trial of dobutamine versus dopamine in preterm infants with low systemic blood flow. AB - OBJECTIVE: Our purpose was to determine if dobutamine or dopamine results in greater improvements in systemic blood flow in very preterm infants with low flow during the first 24 hours of life. STUDY DESIGN: A 2-center, randomized, double blind study. Infants (n = 42) with low superior vena cava (SVC) flow (<41 mL/kg/min) in the first 12 hours were randomly assigned to receive 10 mL/kg normal saline solution, followed by 10 microg/kg/minute of dobutamine or dopamine. If low flow persisted or recurred, the inotrope was increased to 20 microg/kg/minute, with crossover to the other inotrope if treatment failed to maintain flow. RESULTS: Volume produced a more significant increase in SVC flow than dopamine (+43%). At the highest dose, dobutamine resulted in a significantly greater increase in SVC flow than dopamine (mean, +9.9 vs -3.2 mL/kg/min, P =.02). Dopamine resulted in a significantly greater increase in blood pressure. Infants receiving dobutamine only at 24 hours had a greater right ventricular output than infants receiving dopamine (mean, 295 vs 167 mL/kg/min, P <.001). Forty percent failed to increase or maintain SVC flow in response to either inotrope. No significant differences in mortality or morbidity were found. CONCLUSIONS: Dobutamine produced a greater increase in blood flow than dopamine. PMID- 11865270 TI - Preterm infants born at less than 31 weeks' gestation have improved growth in cycled light compared with continuous near darkness. AB - OBJECTIVES: Our purpose was to evaluate the benefits of cycled light (CL) versus near darkness (ND) on health in preterm infants born at <31 weeks' gestational age. STUDY DESIGN: Randomized, interventional study comparing infants receiving (1) CL from birth, (2) CL at 32 weeks' postconceptional age (PCA), and (3) CL at 36 weeks' PCA in transition for discharge home. Statistical significance was assessed with segmented mixed general linear models, analysis of covariance, general estimating equations, chi(2), and Fisher's exact procedure. RESULTS: Infants receiving CL at birth and 32 weeks' PCA gained weight faster than infants not receiving CL until 36 weeks' PCA. There were no differences among the groups in length of hospitalization stay or number of ventilator days, but the power was low for these variables. CONCLUSIONS: These findings suggest that CL has significant weight gain benefits over ND, and there are no short-term advantages of ND over cycled light for health in preterm infants. PMID- 11865272 TI - Lung tissue concentrations of nicotine in sudden infant death syndrome (SIDS). AB - OBJECTIVE: To compare lung concentrations of nicotine and cotinine in cases of sudden infant death syndrome (SIDS) and controls. DESIGN/METHODS: We measured lung tissue concentrations of nicotine and cotinine in SIDS (n = 44) and non-SIDS cases (n = 29) stratified according to household smoking status. RESULTS: When all the SIDS and non-SIDS cases were compared regardless of smoking status, there was a significantly higher nicotine concentration in the SIDS cases than in the non-SIDS cases, (P =.0001). Upon stratifying for smoking status, there was a nonsignificant trend toward more nicotine in SIDS versus non-SIDS lungs that had come from a reported smoking environment. In the nonsmoking group, there were significantly higher nicotine concentrations in SIDS than non-SIDS cases (P =.001). CONCLUSIONS: Children who died from SIDS tended to have higher concentrations of nicotine in their lungs than control children, regardless of whether smoking was reported. These results are based on an objective, biochemical test rather than history, and they further support the relationship between environmental tobacco smoke and the risk of SIDS. PMID- 11865271 TI - Risks of congenital anomalies in large for gestational age infants. AB - OBJECTIVES: To evaluate the association between large for gestational age (LGA) and demographic and medical risk factors as well as specific types of congenital anomalies. STUDY DESIGN: A retrospective, case-control study on 2,149,617 consecutive births was conducted. LGA was defined as 1.64 SD above the mean weight for gestational age, adjusted by sex and altitude. Risk factor frequency distributions were compared between LGA and normal birth weight neonates. Associations between LGA and 41 infants with isolated congenital anomalies were evaluated. RESULTS: Of 31,897 neonates with congenital anomalies, 1800 were LGA. Five anomalies were associated with LGA: talipes calcaneovalgus, hydrocephaly, combined angiomatoses, hip subluxation, and non-brown-pigmented nevi. Multiparity, vaginal bleeding, diabetes, and delivery by cesarean section were more frequent in LGA than in appropriate for gestational age infants' mothers. Several maternal but no paternal factors were statistically associated with an increased risk for LGA infants. CONCLUSIONS: The clinical observation that nevi are more commonly observed in LGA patients was supported. The higher frequencies of hip subluxation and talipes calcaneovalgus among LGA neonates reinforces their pathogenesis as deformations, whereas those of combined angiomatoses and hydrocephaly could reflect increased fluid or body mass. PMID- 11865273 TI - Effects of nutritional status on diarrhea in Peruvian children. AB - OBJECTIVES: We conducted a 4-year (1995-1998) field study in a Peruvian peri urban community (pueblo joven) to examine the relation between diarrhea and nutritional status in 230 children <3 years of age. METHODS: We followed the birth cohort daily for diarrhea and monthly for anthropometry. We modeled diarrheal incidence with a multivariate time-to-event regression model to account for multiple episodes per child and irregular follow-up periods and diarrheal duration with a mixed-effects gamma regression model to account for disease heterogeneity across children. RESULTS: During 159,551 child-days of follow-up, we identified 1387 diarrheal episodes, which yielded an average incidence of 3.2 episodes per child-year. Diarrhea was seasonal, for example, infants had up to 8 diarrheal episodes during the summer; however, these variations decreased noticeably with age. Nutritional status was significantly associated with diarrheal incidence. The frequency of diarrhea increased by 15% per standard deviation decrease in height-for-age z score. Diarrheal episodes in children <6 months of age lasted significantly longer than episodes among older children. CONCLUSIONS: These results identify infants and children of poor nutritional status as priority risk groups for prevention efforts aimed at reducing the burden of acute childhood diarrhea. PMID- 11865274 TI - Allergy to soy formula and to extensively hydrolyzed whey formula in infants with cow's milk allergy: a prospective, randomized study with a follow-up to the age of 2 years. AB - OBJECTIVES: We conducted a prospective, randomized study to evaluate the cumulative incidence of allergy or other adverse reactions to soy formula and to extensively hydrolyzed formula up to the age of 2 years in infants with confirmed cow's milk allergy. STUDY DESIGN: Infants (n = 170) with documented cow's milk allergy were randomly assigned to receive either a soy formula or an extensively hydrolyzed formula. If it was suspected that the formula caused symptoms, a double-blind, placebo-controlled challenge (DBPCFC) with the formula was performed. The children were followed to the age of 2 years, and soy-specific immunoglobulin E antibodies were measured at the time of diagnosis and at the ages of 1 and 2 years. RESULTS: An adverse reaction to the formula was confirmed by challenge in 8 patients (10%; 95% confidence interval, 4.4%-18.8%) randomly assigned to soy formula and in 2 patients (2.2%; 95% confidence interval, 0.3% to 7.8%) randomly assigned to extensively hydrolyzed formula. Adverse reactions to soy were similar in IgE-associated and non-IgE-associated cow's milk allergy (11% and 9%, respectively). IgE to soy was detected in only 2 infants with an adverse reaction to soy. Adverse reactions to soy formula were more common in younger (<6 months) than in older (6 to 12 months) infants (5 of 20 vs 3 of 60, respectively, P =.01). CONCLUSIONS: Soy formula was well tolerated by most infants with IgE associated and non-IgE-associated cow's milk allergy. Development of IgE associated allergy to soy was rare. Soy formula can be recommended as a first choice alternative for infants >or=6 months of age with cow's milk allergy. PMID- 11865275 TI - Effect of hydroxyurea on growth in children with sickle cell anemia: results of the HUG-KIDS Study. AB - OBJECTIVES: Although hydroxyurea is effective in treating adults with sickle-cell anemia (SCA), there is concern that it may adversely affect growth in children. We report the growth characteristics of patients in the Phase I-II pediatric hydroxyurea trial (HUG-KIDS) before and during treatment at the maximum tolerated dose for one year. STUDY DESIGN: Children and adolescents with SCA (n = 68), aged 5 to 16 years at baseline, reached the maximum tolerated dose and had serial height, weight, and Tanner stage measurements. Data from the Cooperative Study of Sickle Cell Disease (CSSCD) were used for comparison. Mixed-effects models were used to compare serial measurements as a function of age and group. RESULTS: In girls, there were no significant differences in height or weight among the pretreatment, on-treatment, and CSSCD groups. Compared with the CSSCD group, HUG KIDS boys were heavier starting at age 9 years, and pretreatment HUG-KIDS boys were taller starting at age 7 years. The Tanner stage transitions took place at appropriate ages. CONCLUSIONS: Hydroxyurea treatment had no adverse effect on height or weight gain or pubertal development in school-aged children with SCA. PMID- 11865276 TI - Dextranomer/hyaluronic acid copolymer implantation for vesico-ureteral reflux: a randomized comparison with antibiotic prophylaxis. AB - OBJECTIVES: Dextranomer/hyaluronic acid (Dx/HA) copolymer has favorable properties for endoscopic treatment of vesico-ureteral reflux (VUR). This open, randomized study was performed to compare the efficacy and safety of Dx/HA copolymer with antibiotic prophylaxis in children with VUR. METHODS: Children >1 year of age with VUR grade II to IV (confirmed by voiding cysto-urethrogram) received endoscopic treatment with Dx/HA copolymer (n = 40) or 12 months of antibiotic prophylaxis (n = 21). Patients in the latter group with reflux grade >or=II at month 3 received a second implantation. All patients were reassessed by voiding cysto-urethrogram at month 12. Scintigraphy and ultrasound were performed to investigate renal status. RESULTS: At month 12, 69% of patients in the Dx/HA copolymer group had reflux grade 1 cm) easily by ultrasonic dissection and a small-caliber pancreatic tube was inserted into the duct on the stump. Subsequently, pancreatic duct invagination could be easily done through a 10 G intravenous catheter passed through the gastrointestinal tract. The main duct was anchored to the adjacent serosa, but any pancreatic parenchymal sutures, possibly leading to internal laceration and/or parenchymal ischemia particularly in soft nonfibrotic pancreases, were avoidable during the procedures. All the anastomoses were done within 10 minutes. Only 1 patient (6.7%)developed pancreatic fistula, which resolved spontaneously in 21 days postoperatively. Neither anastomotic leakage nor remnant pancreatitis was seen in this series. Although a prospective, randomized study is needed, this technique may contribute to reduced morbidity after pancreaticoduodenectomy for a nonfibrotic pancreas with a nondilated main duct. PMID- 11865344 TI - Short-term effects of sacral nerve stimulation for idiopathic slow transit constipation. AB - This study assessed the short-term clinical and physiological effect of continuous sacral nerve stimulation in patients with slow transit constipation. Some patients with idiopathic slow transit constipation are unresponsive to conservative treatments, while colectomy has a variable and poorly predictable outcome. Sacral nerve stimulation is a less invasive and reversible procedure that enables direct neuromodulation of the pelvic floor and hindgut. It has been used successfully in the treatment of urologic disorders and fecal incontinence, and some of these patients with concurrent constipation have also noted improved stool frequency and rectal evacuation. Eight women (median age 47 years, median symptom duration 31 years, median stool frequency once per 6 days) were implanted with a temporary percutaneous stimulating S3 electrode for 3 weeks, attached to an external stimulator (Medtronic, Minneapolis,USA). A bowel symptom diary card, anorectal physiological studies, and a radiopaque marker transit study were completed before and during stimulation. Two patients had cessation or marked diminution of symptoms, including normalization of bowel frequency. Colonic transit did not return to normal in any patient. Rectal sensory threshold to distension was decreased during stimulation. Percutaneous temporary sacral nerve stimulation symptomatically improved a minority of patients with resistant idiopathic slow transit constipation. Sensory function was altered by stimulation. Further studies are required to identify patients who may benefit and to assess a range of stimulation parameters. PMID- 11865347 TI - Structural requirements and interactions of transplant centers. AB - Transplantation of solid organs such as the heart, lung, liver, pancreas, small intestine, and kidney is the only way by which a potentially life-threatening condition can be treated permanently. Transplantation comprises not only the equipment needed to perform the transplantation itself and to offer pre- and postoperative care for the patient but also the know-how of an experienced team of surgeons and all other specialties involved in the care of a transplant candidate (cardiologists, nephrologists, and hepatologists, to name a few). Organ transplantation is a team effort that requires many "wheels to turn" to obtain the successful results we have today. Frequently, enthusiastic individuals try to setup a transplant center "single-handedly." None has succeeded without institutional, financial, and continuous support and in particular without an effectively structured organizational backbone to support the effort. This article summarizes the requirement for a liver transplantation center to perform these procedures and focuses also on ethical considerations. PMID- 11865345 TI - Prospective study of pancreatic b-cell and exocrine function following duct decompression in tropical calcific pancreatitis. AB - Tropical calcific pancreatitis (TCP) is a chronic, nonalcoholic pancreatitis, which is limited to developing countries. In this condition, surgical decompression of the pancreatic duct consistently leads to relief of abdominal pain. However, no data are available on the effect of such intervention on pancreatic function. The aim of the present study was to prospectively evaluate b cell and exocrine function following ductal drainage in patients with TCP. We studied 14 consecutive TCP patients who underwent ductal decompression for abdominal pain (longitudinal pancreaticojejunostomyin 12 patients, endoscopic sphincterotomy and ductal stenting in 2 subjects). Six patients who refused similar intervention served as controls. Patients were evaluated prospectively (median follow-up 13 months) for pain score, fasting and oral glucose stimulated plasma C-peptide, serum trypsin, and fecal chymotrypsin. After intervention, 1 patient died 2 months after surgery, and 2 others were lost in follow-up. The pain score improved significantly following duct decompression (median 8.0 vs. 0, p < 0.01), while in the control group there was no change in pain score (7.0 vs. 7.0). There was no change in b-cell function after intervention (fasting plasma C peptide [mean +/- SEM] 0.41 +/- 0.08 vs. 0.42 +/- 0.05 nmol/l; peak plasma C peptide 2.24 +/- 0.20 vs. 2.32 +/- 0.24 nmol/l). Fecal chymotrypsin was diminished in all patients prior to intervention (1.9 +/- 0.7 U/g), and did not normalize after ductal drainage in any subject. Serum trypsin levels were variable, being elevated in 29% and diminished in 47% of subjects. All 4 subjects with elevated baseline trypsin levels had a sharp fall after intervention (1020 vs. 175 ng/ml). However, serum trypsin did not normalize after ductal drainage in any patient with a diminished baseline value. In conclusion, patients with TCP have significant reduction in abdominal pain after decompression of the main pancreatic duct. However, there is no significant change in b-cell function. A fall in elevated serum trypsin suggests that there may be relief of subclinical inflammation after intervention; however, there is no improvement in exocrine function after a follow-up of 1 year. PMID- 11865348 TI - Organ donors: heartbeating and non-heartbeating. AB - The limits of organ donation from heart-beating (HB) donors reached a plateau illustrated by the number of postmortem kidneys for transplantation. Programs such as the European Donor Hospital Education Program (EDHEP) and Donor Action have helped to stop a further decrease in the number instead of an expected increase. For kidneys, heart, liver, and lungs one must also explore the use of marginal donors as a possible additional source. Examples are donors with a horseshoe kidney, those at both ends of the age spectrum, and those with medical contraindication such as diabetes. We have enlarged our kidney donor pool considerably with non-heart-beating(NHB) donors. Because we preserve these kidneys in a preservation machine, we are able to perform viability testing. With glutathione S-transferase (GST) as a measure of tubular damage, we now decide whether to transplant based on GST values. For other organs, NHB donation does not seem to be an option other than for the liver when the warm ischemia time is short. PMID- 11865349 TI - Kidney transplantation: graft monitoring and immunosuppression. AB - Renal transplantation has become the preferred means of treating end-stage renal disease. Episodes of allograft rejection have become the exception rather than the rule. The development of real-time ultrasound-guided allograft biopsy and adoption of the Banff criteria for histologic evaluation permit safe,accurate monitoring of graft histology. New immunosuppressive agents have drastically reduced the number of episodes of both primary and refractory rejection. Novel biologic agents in the form of monoclonal antibodies and soluble receptor hybrid molecules may serve to reduce the required doses of toxic chemical immunosuppressants and provide more specific immune suppression directed at those elements of the immune system involved in rejection of a given allograft. Development of assays to identify patients who demonstrate donor antigen-specific hyporeactivity is now feasible. Hopefully, these assays will serve as a guide for the reduction and possible removal of immunosuppressive agents from stable renal allograft recipients. PMID- 11865350 TI - Technical and immunosuppressive advances in transplantation for insulin-dependent diabetes mellitus. AB - Pancreas transplantation has emerged as the single most effective way to achieve normal glucose homeostasis inpatients with type I insulin-dependent diabetes mellitus. Optimal immunosuppressive strategies for pancreas transplantation continue to evolve with the use of newer, more potent immunosuppressive agents,particularly tacrolimus, mycophenolate mofetil, and rapamycin. These agents have contributed to substantially lower rates of allograft rejection and improved graft survival. Regimens designed to avoid nephrotoxicity or spare corticosteroid therapy are emerging as the variety of drug options grows. Also contributing to progressively better results for solitary pancreas transplants are reductions in early graft loss rates and the development of safe, effective biopsy techniques, permitting accurate diagnosis of rejection. Collectively,these factors have allowed solitary pancreas allograft recipients, a group of patients with historically poor long-term graft survival, to enjoy successes nearly equivalent to those of combined kidney-pancreas transplants. Consequently, the American Diabetes Association strongly endorses pancreas transplantation in diabetic patients who have received prior kidney transplants and who have life threatening metabolic lability. PMID- 11865351 TI - Bridging procedures to heart transplantation. AB - Ventricular assist devices are used to support the failing circulation and consequently bridge patients with end-stage heart disease to heart transplantation. From 1988 through 2000 we mechanically supported 47 patients with a bridge to heart transplantation. Within the same time frame 118 patients were mechanically supported for recovery of the heart. Most presented in acute cardiac failure, and the severe shock leads to high early mortality. Mortality during mechanical support is 36%. During the early experience patients underwent transplantation urgently. With the more recent implantable devices, patients are fully mobilized and given transplants electively. Long-term survival of these patients bridged to transplantation is excellent and does not differ from that for non-bridged patients (5-year survival is 82% for the bridged patients and 84% for the non-bridged patients; p = 0.43). The most frequent device-related problem is excessive bleeding (38%). Thromboembolic phenomena are the most cumbersome complications (11%). The strategy of bridging to heart transplantation has evolved from acute resuscitation and urgent transplantation to medium- and long term support to optimize the patient's condition. New experience with long-term support and alternative support strategies has created possibilities in the field of "alternatives to heart transplantation." PMID- 11865352 TI - Heart and heart-lung transplantation: standards and improvements. AB - Standards and new developments of thoracic organ transplantation are reviewed with particular focus on current treatment strategies, alternatives to transplantation, and xenotransplantation. The current indications for heart, single and bilateral sequential lung, and heart-lung transplantation as well as the technical aspects of each procedure are presented. Criteria for transplant recipients and absolute and relative contraindications are pointed out. Criteria for donor selection are also reviewed. The results of single, double-sequential, and heart-lung transplantation over the past 10 years as reported by the International Society for Heart and Lung Transplantation Database are stated. In addition, the experience of the lung and heart-lung transplantation program at the Hannover Medical School is reviewed, including the current immunosuppression regimens. This experience includes 1075 heart,heart-lung, and lung transplantations since 1983. The 1- and 5-year actuarial survival rates for heart transplant recipients are 81% and 70%, for heart-lung recipients 76% and 61%, and for single and double lung transplant recipients 77% and 59%, respectively. During the past decade there has been continuous improvement in the results of heart, lung, and heart-lung transplantation. Alternatives to thoracic organ transplantation, living-related lobar transplantation, new antirejection agents, and xenograft transplantation are areas for continuing and future investigation. PMID- 11865353 TI - Intestinal and multivisceral transplantation. AB - Intestinal transplantation has been gradually instituted in the management of intestinal failure. More than 200 cases including isolated intestinal transplant, liver/intestinal transplant, and multivisceral transplant have been performed worldwide,with 1-year graft and patient survival rates of 66% and 54%,respectively. Indications for the procedure include short bowel syndrome and functional abnormalities secondary to a variety of diseases or conditions. Tacrolimus-based immunosuppression regimens have been used universally with improved outcomes. The major contributors to the morbidity and mortality include rejection,infection, and technical complications. Of those, control of rejection remains the most difficult dilemma and it will be the key to improved patient and graft survival. PMID- 11865354 TI - Living unrelated kidney transplantation. AB - The use of living donors, particularly if unrelated, in kidney transplantation is still not recommended although many transplant centers have come to accept the procedure. Usually the main argument against this approach is ethical. Acknowledging this problem, we accept biologically unrelated donors only if they have an emotional closeness to the recipient. From November 1982 to November 1997, 527 kidney allografts from living donors were performed at our institution. Of these 302 living donors were first-degree relatives of the recipient and shared one haplotype (LRD) and 172 were unrelated(LURD). Among the LURD group 146 donors were "emotionally related"--wife to husband 110 cases and husband to wife 35 cases and 1 case from a nun to a friar. Statistical analysis of the results was performed with the chi(2) method. Actuarial graft survival rates in the LRD and LURD groups were 91% and 87% at 1 year, 77% and 79% at 5 years, and 66% and 69% at 9 years (P =n.s.). In conclusion kidney transplantation between unrelated donors and recipients may be a valid alternative in view of the cadaver organ shortage. It is a procedure that can be performed successfully and that provides a "gift of life" for both the patient and the family. PMID- 11865355 TI - Recent advance in living donor liver transplantation. AB - Living donor liver transplantation (LDLT)has been performed in more than 2000 cases around the world. This procedure is considered to have certain advantages over cadaveric liver transplantation, because detailed preoperative evaluation of the donor liver is possible and superior graft quality is available. The indication has recently been widened to include adult patients. The results of LDLT have been reported to be very good. In this article,several considerations on LDLT,including living donor selection and application to adult patients, are discussed. Between June 1990 and March 2001, 143 patients underwent LDLT at Shinshu University Hospital. During this period, 160 patients were determined to be candidates for liver transplantation in our institution, and 185 candidates were evaluated as potential donors for these patients. Thirty-eight of 185 donor candidates were excluded for reasons including liver dysfunction and withdrawal of consent. The recipients included 60 adults, 50 (83%) of whom are currently alive. Taking into account the worldwide shortage of cadaveric organ donation,the importance of LDLT will probably never diminish. This procedure should be established on the basis of profound consideration of donor safety as well as accumulated expertise of hepatobiliary surgery. PMID- 11865356 TI - Liver transplantation for nonviral, nonmalignant diseases:problem of recurrence. AB - Liver transplantation has evolved rapidly from an experimental treatment to universally accepted therapy for end-stage liver disease. Indications for liver transplantation have expanded with the evolution of the procedure to include metabolic,viral, malignant, and acquired liver failure. As long-term liver transplant recipient follow-up data become available, the development of recurrent liver disease within the transplanted allograft is an increasing dilemma. The value of the transplant procedure must be assessed within the context of survival as well as the potential for recurrent disease and the associated need for re-transplantation. This study represents a compilation of data obtained from adult patients undergoing liver transplantation for nonviral, nonmalignant etiologies at the University of California San Francisco and a comparison of our data with other centers. Between February 1988 and January 1997, 654 liver transplants were performed on 623 patients. From this group, 406 recipients were identified as meeting study inclusion criteria: age above 18 years, and transplantation for a nonmalignant, nonviral etiology of liver failure. Indications for liver transplantation included primary biliary cirrhosis (n = 65), primary sclerosing cholangitis (n = 49), autoimmune hepatitis (n = 37),Budd-Chiari syndrome (n = 7), cryptogenic cirrhosis (n = 88),and alcoholic liver disease (n = 160). Mean follow-up within the diagnostic groups ranged from 4.9 to 5.6 years. Evaluation of clinical,immunosuppressive, and pathologic data for each diagnostic group was performed to determine the incidence, time to recurrence, clinical presentation, and sequelae of disease recurrence. PMID- 11865357 TI - Liver transplantation for malignant diseases: selection and pattern of recurrence. AB - Liver transplantation (LT) for malignant tumors should be accepted if, with adequate case selection, long-term results are similar to those in patients transplanted for benign diseases. The aim of the present study was to reexamine selection criteria for LT in malignant diseases with particular emphasis on hepatocellular carcinoma (HCC) in cirrhosis. One hundred-three of 369 patients transplanted in our unit had HCC in cirrhosis (28%), 15 of which were incidental tumors, and 234 patients underwent LT for non-cholestatic cirrhosis. Pretransplant arterial chemoembolization(TACE) was performed in 36 cases (41%) of known HCC. Only early,well-delimited tumors in advanced cirrhosis with no extrahepatic disease were accepted for LT. Hepatocellular carcinoma characteristics included mean tumor size (3.1 cm), multiple (59%), bilobular involvement (31%), and vascular invasion (9.2%). Postoperative mortality was 4%. Median follow-up was 67.5 months. Tumor recurrence rate was 14.5%, 33% (5/15) in incidental tumors and 11.4% (10/88) in known HCC and by tumor stage (pTNM): 7.7% (1/13) in stage I, 16.7%(5/30) in stage II, 15% (3/20) in stage III, and 17% (6/35) in stage IV. Mean time for recurrence was 20.6 months. Tumoral vascular invasion, tumor differentiation, and satellite tumors were significant factors for tumor recurrence in univariate analysis, whereas tumor vascular invasion was the only significant factor for tumor recurrence in multivariate analysis. Actuarial survival rates at 1, 3, and 5 years were 81%, 66%, 58%, respectively, in patients with HCC and were similar to those of cirrhotic patients 76%, 67%, 63%, respectively. In conclusion, patients with early HCC in cirrhosis are good candidates for LT; results are similar when compared with those of cirrhotic patients without tumor. Liver transplantation for other malignancies is admitted only in fibrolamellar hepatoma, hepatoblastoma, epithelioid hemangioendothelioma without extrahepatic disease, and in metastases from carcinoid tumors. PMID- 11865359 TI - Split-liver transplantation: future use of scarce donor organs. AB - The main obstacle to a expansion of human liver transplantation is the lack of donor organs. At present mortality reported for pediatric and adult patients on the waiting list is 10%to 20%. This article focuses on several techniques to alleviate this problem. Several years ago, application of reduced-size liver transplantation overcame the donor shortage among small infants through the use of grafts shaped to almost any size needed. Today reduced-size grafts are only rarely used, most commonly with traumatized donor livers or particularly small pediatric donor livers. Split liver transplantation also yields a net gain of organs, in that it uses one organ to save either an adult and a child or, recently, two adults. The technique of ex situ splitting is progressively being replaced by the in situ splitting technique, which yields better preserved grafts,optimization of graft/donor matching by pretransplant manipulation(preconditioning), avoidance of early rejection in the recipient,portal decompression, temporary liver support, if necessary, and induction of fast regeneration. In acute hepatic failure, auxiliary heterotopic liver transplantation might be sufficient to support liver function until regeneration of the native liver has begun. Domino transplantation in some patients with inborn errors of metabolism or storage disease should be considered. This article focuses on increasing the organ supply by using split liver transplantation techniques and living-donor liver transplantation. PMID- 11865358 TI - Auxiliary liver transplantation and bioartificial bridging procedures in treatment of acute liver failure. AB - The aim of this review is to discuss the place of auxiliary liver transplantation (ALT) and other cellular based bridging procedures such as hepatocyte transplantation, ex vivo liver perfusion, and bioartificial livers, in the treatment of acute liver failure, vis a vis conventional orthotopic liver transplantation. Hepacocyte transplantation, and ex vivo pig or human liver perfusion are still experimental procedures. Bioartificial livers using human tumoral hepatocytes or porcine hepatocytes have been used in clinical situations as a bridge to transplantation, i.e. to gain the time required to find a high quality graft for conventional or auxiliary liver transplantation. None of these techniques have yet proved capable of keeping a patient alive long enough for the native liver to recover. Conversely, ALT has been shown to be effective in the treatment of acute liver failure and now appears to be a satisfactory bridging procedure pending native liver (NL) regeneration. We report personal experience of 18 ALT procedures performed in 17 patients between October 1992 and December 1999. The ALT procedure was indicated when patients met criteria for conventional transplantation; it was ultimately selected when a fresh frozen biopsy of the NL did no show any fibrosis. Six patients died within the first 2 postoperative months. The remaining 11 patients are alive, with a follow-up ranging from 2 to 7 years. Regeneration of the NL occurred in 11 of the 17 patients (65%) and in 8 of the 11 survivors (72%), 6 of whom have permanently stopped immunosuppressive therapy. We conclude that liver failure should no longer be handled outside centers where all types of transplantation can be offered, and where innovative therapies such as hepatocyte transplantation and extracorporeal liver-assist devices are being developed and evaluated. PMID- 11865362 TI - Critical analysis of the treatment of non-palpable breast cancer: toward a less invasive future? AB - Results and experience with the treatment of non-palpable breast cancer are reported and related to the future of less invasive treatment of small breast cancers. The retrospective study included 102 patients treated between 1980 and 1993 at the University Hospital of Nijmegen. Most cancers were screen-detected. Tumor sizes on pathological examination proved 3 mm (SD 7.7 mm) larger than on mammography (p = 0.0029). pT1 tumors were encountered in 77 patients (75%); pT2 tumors in 17 patients (17%). Seventy-five patients were free of lymph node metastases, 26 patients showed stage pN1 (25%). Most patients had invasive ductal cancers. Only two patients died of breast cancer. A 10-year disease-free survival of 94% was calculated, after excluding four patients with known systemic disease (M1) at diagnosis. T1a, T1b, and T1c had a 10-year survival of 100%, 96%, and 96%, respectively. Early detection and multimodality treatment of breast cancer have significantly improved survival. Inpatients with small breast cancer tumors future developments in treatment must be aimed at the use of less invasive techniques, reducing morbidity while maintaining high levels of disease-free survival. PMID- 11865363 TI - Breast microcalcifications: multivariate analysis of radiologic and clinical factors for carcinoma. AB - Screening mammography contributes to the improvement of breast carcinoma survival through early detection and treatment of non-palpable lesions. Microcalcifications are of fundamental importance in this process. The percentage of malignant lesions found in biopsies for microcalcifications varies from 10% to 40%. The purpose of this study was to evaluate the relationship between clinical and radiologic records and the presence of malignant breast diseases. To establish the basis for the study, 211 mammographic files showing clustered microcalcifications from 204 women were prospectively reviewed and clinical records were retrospectively drawn. Definitive pathologic analysis was available for all. The value for cancer of each criterion was investigated by univariate and multivariate analyses. A first analysis was performed on the entire population and a second one was performed with stratification on morphologic subgroups. There were 99 malignant lesions (47%). In the entire group, no clinical criterion was significant. In the univariate analysis, five radiologic variables were significant: morphologic type(p < 0.0001), number of calcifications per cluster(p < 0.0001), linear or triangular distribution(p < 0.0002), diameter of the area (p < 0.01),and number of clusters (p = 0.011). In the multivariate analysis, two criteria remained significant: morphologic type 4 (irregularly punctiform) or 5 (vermicular) microcalcifications(Le Gal's classification) (p < 0.0001) and diameter of the cluster larger than 25 mm (p = 0.032). In subgroups,in the multivariate analysis, the "age > 60 years" criterion was statistically significant in the group of regular punctiform microcalcifications (type 2); for irregularly punctiform microcalcifications (type 4), "number of microcalcifications > 20" was significant. The morphologic features of microcalcifications must be the first criterion evaluated. They permit identification of characteristically benign (annular calcifications) or malignant calcifications (vermicular calcifications). For the remainder of the calcification types, other criteria must be taken into account, and their value vary with (according to) the morphologic aspect. These findings have implications for the management of women with microcalcifications and could help breast specialists make treatment decisions. PMID- 11865364 TI - Influence of exogenous fat emulsion on pulmonary gas exchange after major surgery. AB - The use of fat emulsion in total parenteral nutrition (TPN) is closely related to changes in respiratory function. The aim of this study was to evaluate the influence of exogenous fat emulsion on pulmonary gas exchange in the early period after major surgery. Total parenteral nutrition was administered to 18 patients for 6 days after esophagectomy for carcinoma. Half of the patients received glucose (glucose group), and the other half received glucose and fat (fat group). The fat emulsion was continuously infused for 24 hours over 6 days. Glucose utilization was significantly higher in the glucose group than in the fat group. Fat utilization was significantly higher in the fat group than in the glucose group. Carbon dioxide (CO2) production and respiratory quotient were significantly decreased in the fat group compared to the glucose group. There were no differences in the pulmonary vascular resistance index or alveolar arterial difference in oxygen tension between the two groups. Although exogenous fat emulsion utilized as energy substrate decreases CO2 production after major surgery, it does not clinically influence the pulmonary hemodynamics or diffusion capacity. PMID- 11865365 TI - Influence of postoperative epidural analgesia with bupivacaine on intestinal motility, transit time, and anastomotic healing. AB - Epidural application of bupivacaine has been suggested to have a sympatholytic effect on spinal reflex mechanisms that shortens postoperative paralysis and leads to an improved transit time. The influence on anastomitic healing remains controversial. Laparotomy was performed in eight dogs. A short segment of the distal colon was resected and five electrodes were fixed on the serosa to measure the myoelectric activity (e.g., Migrating Myoelectric Complex--MMC). After operation a peridural catheter was placed between L7 and the sacral crest. One milliliter of bupivacaine 0.25% for each 3 kg of body weight was injected every 4 hours. Barium pellets coated in wax were placed into the stomach to allow radiographic representation of transit time. After 5 days the colon anastomosis was resected to measure the bursting pressure. In the peridural analgesia group (PDA) we found one small bowel intussusception and one covered anastomotic leakage. Postoperative PDA led to early and severe myoelectric activity but did not influence the time until the first MMC occurred (44 +/- 0.8 h, PDA; 44.6 +/- 1.5 h,control). Neither the transit time to the colon (50.2 +/- 1.9h, PDA; 51.7 +/- 5.5 h, control) nor the anastomotic healing was influenced (bursting pressure: 176 +/- 21.1 mmHg, PDA; 152 +/- 27.7 mmHg, control). Postoperative epidural analgesia with bupivacaine shortens intestinal paralysis. Early myoelectric activity with a lack of propulsive activity can cause complications like small bowel intussusception. Hence early postoperative enteral nutrition after epidural analgesia is risky. Because the influence of epidural analgesia on propulsive motility remains unclear, it seems reasonable to recommend its limited use in colon surgery. PMID- 11865366 TI - Surgery and adjuvant therapy in patients with diffuse peritonitis: cost analysis. AB - In a prospective, randomized, controlled trial the effect of high dose intravenous antithrombin III and intraabdominal donor serum was analyzed in 36 patients with diffuse secondary peritonitis. The direct cost for treatment was 25,370 euros per patient, and the post acute hospital care costs and societal costs were 6273 euros. The cost for intensive care of these patients accounted for approximately 83% of the direct costs, while the expenditures for operating theater and general wards accounted for 9% each. The most expensive factors were staff, medication, and blood products. The hospital incurred a deficit of 3696 euros for each patient after reimbursement from public health insurance companies. Quality of life as assessed by the gastrointestinal quality of life index (GIQI) showed a good outcome. On average 11 quality adjusted life years (QALY) were achieved. The cost per QALY was 2631 euros. Use of adjuvant therapy was associated with a reduced duration of intensive care unit (ICU) treatment, times on mechanical respiration, and hemofiltration; the cost of treatment was reduced by 6614 euros per patient. The additional cost of antithrombin III (5155 euros) was more than offset by the savings made when adjuvant therapy was used. PMID- 11865367 TI - Cases with fewer than four parathyroid glands in patients with renal hyperparathyroidism at initial parathyroidectomy. AB - In the surgical treatment of secondary hyperparathyroidism (2HPT) due to uremia, it is considered necessary to remove all parathyroid glands from the neck to prevent persistent and recurrent parathyroid hyperfunction. However, in some cases fewer than four parathyroid glands can be recognized at initial operation; in the present study, we evaluated the long-term prognosis and estimated surgical strategy in such cases. Between March 1981 and January 1999, 822 patients underwent total parathyroidectomy (PTx) with forearm autograft for advanced 2HPT at the Department of Transplant Surgery of Nagoya Second Red Cross Hospital. In 21 cases (2.6%) fewer than four parathyroid glands were macroscopically found at the initial operation. These cases were followed up and their parathyroid function was evaluated by measurement of intact parathyroid hormone (PTH). In 20 of the 21 cases three glands were found, in 1 patient only two glands. In 5 of these cases the fourth gland was identified first after postoperative histopathologic evaluation. In all these cases the intact PTH level was normalized. In 8 of the remaining 16 cases high PTH levels persisted after the initial operation, including 3 patients who underwent neck reexploration. However, in the other 7 patients PTH levels dropped within normal range immediately after PTx and a fourth gland has never been recognized. One patient was lost to follow-up. Thus, using our operative strategy, 12 of 822 cases (0.85%) did not develop persistent or recurrent HPT even though only three glands were identified at the operation. To avoid postoperative hypoparathyroidism, autotransplantation should be performed when fewer than four parathyroid glands are found at the initial operation. PMID- 11865368 TI - Sentinel node concept in gastric carcinoma. AB - To assess the applicability of the sentinel node concept to gastric carcinoma. The location of metastatic lymph nodes was analyzed retrospectively in 119 patients with gastric carcinoma in whom metastasis was limited to one or two nodes. Intraoperative lymphatic mapping was attempted in 62 patients using indocyanine green injected endoscopically into the gastric submucosa adjacent to the tumor. Metastatic lymph nodes were distributed beyond the perigastric area in 4% of patients with a single node involved. The positive node was located along the greater curvature in 21% of the patients with a tumor on the lesser curvature. Two patients had a metastatic node totally occupied by cancer tissue. In 16% of patients with two nodes involved, a positive node was located on both the lesser and greater curvatures. Lymphatic mapping was successful in all subjects. A larger number and wider distribution of green-stained nodes were observed in patients injected with 8 ml of indocyanine green solution than in those injected with 4 ml. No metastasis was observed in any nodes in 47 (96%) of the 49 patients who had no metastasis in green nodes. In one patient showing metastasis in non-green nodes without metastasis in green nodes, the positive nodes were totally occupied by cancer tissue. Our results showed the complexity of lymphatic streams within and from the stomach. Lymphatic mapping using indocyanine green can be a tool for identifying sentinel nodes in gastric carcinoma although lymph nodes occupied by cancer tissue may not be detected by this technique. PMID- 11865369 TI - Prognostic significance of metastatic lymph node ratio in T3 gastric cancer. AB - The fifth International Union Against Cancer tumor node metastasis (UICC TNM) classification, based on the number of metastatic lymph nodes (LN), has proved to be a reliable and objective method for predicting the prognosis of patients with gastric cancer. However, the prognosis of patients with T3 gastric cancer is still heterogeneous. This study was carried out to investigate the validity of metastatic LN ratio as a prognostic factor in T3 gastric cancer. A retrospective analysis was performed on a total of 833 patients that had either T3N1M0 (n = 504) or T3N2M0 (n = 329) gastric cancer by the fifth UICC classification. A preliminary analysis revealed the cutoff values for T3N1M0 to be 10% and for T3N2M0 to be 25%. The mean metastatic LN ratio was 9.0% for T3N1M0 cancer and 26.9% for T3N2M0 cancer. For the T3N1M0 stage, the patients who showed less than 10% of the metastatic LN ratio were grouped as N1-low with the others grouped as N1-high. For the T3N2M0 stage group, those who had less than 25% of the metastatic LN ratio were grouped as N2-low, the remainder as N2-high. The metastatic LN ratio decreased in proportion to the extent of lymphadenectomy and it increased in relation to the increasing scale of the fourth N classification. The rates of recurrence were significantly different according to the metastatic LN ratio in N1 and N2 classification of the fifth UICC classification (p < 0.05). The 5-year survival rates after gastrectomy decreased significantly by increasing the metastatic LN ratio in both T3N1M0 cancers (p =0.0026) and T3N2M0 cancers (p = 0.0057). The metastatic LN ratio was an independent risk factor for recurrence and poor prognosis. Our data suggest that the metastatic LN ratio is a significant prognostic factor for T3 gastric cancer. Furthermore, the application of the metastatic LN ratio can provide information not only about the extent of LN metastasis but also about the extent of lymphadenectomy in T3 gastric cancer. PMID- 11865371 TI - Quality of life after gastrectomy: Longmire's reconstruction alone compared with additional pouch reconstruction. AB - The optimal reconstruction protocol after total gastrectomy is still a matter of debate. Pouch reconstructions are developed to create a larger reservoir for food, to provide a barrier against intestinoesophageal reflux, and to lengthen the food transit time. Preservation of the duodenal passage should result in better physiologic regulation of the ingested food. Controlled randomized clinical studies must be conducted to assess the quality of life after gastrectomy and various types of reconstruction. In the present trial, which compared Longmire's reconstruction without a pouch and Longmire's reconstruction with a pouch of varying sizes, we evaluated the quality of life for 41 patients during the first postoperative year. The quality of life was examined by an EORTC questionnaire and an organ-specific module. There were no statistically proven benefits for pouch reconstruction in comparison with Longmire's reconstruction alone. Some benefits are shown for the large pouch but it was only a trend. This is especially so with regard to diarrhea and food consumption. A definitive answer to the question about the value of a pouch reconstruction after gastrectomy has not yet been attained. It is possible that there will be significant differences between reconstruction groups during the long-term follow up. PMID- 11865370 TI - Early gastric cancer: ten years of experience. AB - Gastric cancer is a disease in which the main treatment is surgical extirpation. The modifications introduced in the surgical treatment over the last decades were accompanied by a clear increase of survival, which reaches global values of 61% at 5 years in Japan. One of the reasons that contribute to this improvement is early diagnosis of the lesions. In the period between January 1, 1990 and December 31, 1999 662 patients with gastric adenocarcinoma were treated in the Service of Surgery 1 of our hospital; 110 were refused surgical treatment. Of the resected patients, 91 (21.4%) were classified as early gastric cancer according to the definition of the Japanese Society of Digestive Endoscopy. There were 30 women and 61 men, with a median age of 60.2 +/- 15 years; 3 patients had a preoperative diagnosis of gastric ulcer; 2 others were operated without recent histology; and 1 patient was urgently resected for a bleeding ulcer. In all the remaining patients biopsy confirmed the presence of cancer (89%) or serious dysplasia (4.6%). The lesions had been distributed essentially in the medium 1/3 (48.3%) and distal 1/3 of the stomach. Subtotal gastrectomy was accomplished in 48 patients, total gastrectomy in 40, total desgastrogastrectomy in 3, and in 9 patients the surgery involved the spleen (8 patients) and the spleen and tail of the pancreas in 1 patient. Lymphadenectomy was not performed in 5 patients, lymph nodes by the first lymph node barrier were removed in 25 patients and by the second barrier in 61 patients (67%). Median tumor size was 26 +/- 1.8 mm. The lesion reached the mucosa in 46 patients and the mucosa and submucosa in 45. In 6 patients the removed lymph nodes were microscopically invaded (6.7%). Five patients died (5.7%). The median follow-up of the patients is 41 +/- 26 months; 7 patients died (8.1%) during this period; 4 died unequivocally of disease progression. The median survival of patients was 85% at 5 years and 80% at 10 years. In our series, survival was affected by the presence of invaded lymph nodes, not by the penetration in depth of the lesion or the size of the tumor. PMID- 11865372 TI - Results of end-to-end cavocavostomy during adult liver transplantation. AB - The results of end-to-end cavocavostomy during adult liver transplantation were analyzed with special regard to caval complications. In a series of 1000 liver transplants, we observed 17 patients who suffered from postoperative caval obstruction (6 patients) or caval stenosis (11 patients), for an incidence of 1.7%. Surgical therapy was performed in 10 patients (58.8%), and 5 patients required retransplantation (29.4%). Four patients died during the later postoperative course. Two fatalities were related to caval complications, resulting in a mortality rate of 11.8%. Our results indicate that end-to-end cavocavostomy is a safe technique for cavocaval anastomosis. For only a few exceptions, such as pediatric transplantation, reduced size livers, or size mismatch between donor and recipient, should alternative techniques such as end to-side or side-to-side cavocavostomy be performed. PMID- 11865373 TI - Intraoperative iatrogenic rupture of hepatocellular carcinoma. AB - Intraoperative iatrogenic rupture of hepatocellular carcinoma (HCC), which can occur during hepatic resection when large tumors are being mobilized, may adversely affect the operative outcome. Little information is available in the literature on this serious intraoperative complication. The aim of the present study is to document iatrogenic rupture of HCC as a serious complication during hepatic resection and its effects on the operative and long-term outcomes of patients with this complication. A retrospective study was performed on all patients with intraoperative iatrogenic rupture of HCC during hepatic resection from 1989 to 1997, and the operative and long-term survival outcomes were compared with those of patients without the complication. Among 194 patients who underwent hepatic resection for a large HCC (> or =5 cm) during the study period, 8 (4.1%) had intraoperative iatrogenic rupture of the tumor. When compared with 186 patients with similar clinical parameters but without intraoperative rupture, patients with intraoperative rupture had significantly more intraoperative blood loss (median 5.7 vs. 2.0 L;p = 0.01) and blood transfusion requirement (median 3.1 vs 0.9 L; p = 0.02). On follow-up, patients in the intraoperative rupture group had a significantly higher intraperitoneal extrahepatic recurrence rate (33.3% vs. 2.9%; p =0.02) and significantly shorter survival (median 11.5 vs. 37.9 months,p = 0.04) when compared with patients without the complication. Intraoperative iatrogenic rupture is a serious complication of hepatic resection for HCC because it is associated with increased intraoperative blood loss, increased incidence of intraperitoneal extrahepatic recurrence, and short survival. Extreme care should be taken during mobilization of the tumor, and an alternative operative approach in the presence of a difficult hepatic resection of a large HCC may be required to avoid the complication. PMID- 11865374 TI - Efficacy of diagnosis of mechanical cholestasis by magnetic resonance cholangiography. AB - The aims of this study were to evaluate the effectiveness of magnetic resonance cholangiography (MRC) in diagnosing patients with cholestasis and to compare these results to those obtained with endoscopic retrograde cholangiopancreaticography(ERCP). From January 1996 to December 1998 a total of 85 consecutive patients who were candidates for ERCP because of a suspected diagnosis of mechanical cholestasis were included in this study. All patients underwent MRC and consecutive ERCP 24 to 48 hours later. The sensitivity for detecting common bile duct stones was 93%,specificity 74%, positive predictive value 89%, and negative predictive value 82%. For the determination of tumorous bile duct stenosis the sensitivity and specificity were both 100%. MRC might be an ideal method for evaluating patients with unclear laboratory or ultrasonography findings before laparoscopic cholecystectomy, thereby avoiding unnecessary invasive diagnostic tests with possible harmful complications. PMID- 11865375 TI - Inhibitory effect of loxiglumide (CR 1505), a cholecystokinin receptor antagonist, on N-nitrosobis(2-oxopropyl) amine-induced biliary carcinogenesis in Syrian hamsters. AB - We evaluated the cholecystokinin (CCK) receptor antagonist loxiglumide (CR1505) for a possible inhibitory effect on biliary carcinogenesis in a hamster model. Experimental group I underwent cholecystoduodenostomy and ligation of the distal end of the common bile duct, after which the animals were injected with N nitrosobis(2-oxopropyl)amine (BOP) alone. Group II, after the same surgical procedure as in group I, were given injections of BOP and then given loxiglumide in their diet. The sham-operated group underwent simple laparotomy and then were given injections of BOP. Loxiglumide significantly inhibited BOP carcinogenicity in the gallbladder and extrahepatic bile duct but not in the intrahepatic bile ducts or pancreas. Autoradiography showed that loxiglumide significantly suppressed (125)I-Bolton-Hanter (BH)-CCK-8 binding to CCK receptors in the gallbladder and extrahepatic bile duct but not in the liver or pancreas, and CCK binding to its receptors was observed in an area identified as cancer tissue. CCK receptor antagonists have an inhibitory effect on BOP carcinogenesis in the extrahepatic biliary tract, including the gallbladder and extrahepatic bile duct, of Syrian hamsters. The difference in the inhibitory effect of loxiglumide on biliary carcinogenesis in hamsters according to site may be due to differences in CCK receptors or the affinity of loxiglumide for such biliary tract organs. A difference between carcinogenesis in the intrahepatic bile ducts and extrahepatic biliary tract may be another reason. PMID- 11865376 TI - Comparison of the functional outcome after pylorus-preserving pancreatoduodenectomy: pancreatogastrostomy and pancreatojejunostomy. AB - To determine if there is any difference in pancreatic function after pylorus preserving pancreatoduodenectomy(PPPD) according to the type of pancreatoenterostomy [pancreatojejunostomy (P-J) or pancreatogastrostomy (P-G)], we evaluated the long-term functional status of 34 patients who underwent PPPD and survived for more than 1 year without clinical evidence of recurrence. Altogether 20 patients underwent P-J and 14 P-G. To compare the two groups, we analyzed the (1) general nutritional status; (2) quality of life using three scoring systems; (3) gastrointestinal symptoms; and (4) pancreatic exocrine function by the stool elastase I test and endocrine function by oral glucose tolerance test (GTT). After PPPD, body weight decreased in both groups, with no difference between the two groups. No statistical differences were found in triceps skinfold thickness or serum protein/albumin. Regarding the quality of life and postoperative gastrointestinal symptoms, there were no differences between the two groups except steatorrhea. There were 4 mild and 15 severe cases of pancreatic exocrine insufficiency among those who underwent P-J, whereas all of the patients who underwent P-G showed severe pancreatic insufficiency. On GTT, excluding preoperative diabetes patients, 43.8% (7/16) of the P-J group had abnormal results after surgery, whereas 75.0% (9/12) of the PG group had an abnormal postoperative GTT (p = 0.11). Severe exocrine and endocrine pancreatic insufficiency developed after PPPD in both the P-J and P-G groups, but there was more functional deterioration in the P-G group than in the P-J group. General nutritional status and quality of life were not affected by the pancreatoenterostomy method in either group. PMID- 11865377 TI - Characteristics of infection with Candida species in patients with necrotizing pancreatitis. AB - This study focuses on the relevance of Candida infection (albicans and non albicans) in patients with necrotizing pancreatitis. Altogether, 92 patients with infected pancreatic necrosis were reviewed for Candida infection. All patients underwent surgical necrosectomy for infected pancreatic necrosis. Data from patients with Candida growth in intraoperative smears were compared to those obtained from patients without Candida infection. There were 22 patients (24%) with Candida infection. Patients with or without Candida infection were comparable regarding age, gender, etiology, and severity scores at admission. Candida patients suffered a higher mortality (64% vs.19%, p = 0.0001) and experienced more systemic complications (3.2 +/- 1.6 vs. 2.1 +/- 1.4; p= 0.004) than patients without Candida. Preoperative antibiotics were given significantly longer prior to Candida infection (19.0 +/- 13.2 vs. 6.4 +/- 10.3 days; p < 0.0001). With regard to the concomitant spectrum of bacteria, solitary gram negative infection was rare in Candida patients (5% vs. 43%, p =0.0006). The presence of Candida in patients with infected pancreatic necrosis is associated with increased mortality. Our data provide evidence that application of antibiotics contributes to the development of Candida infection and to changes in the bacterial spectrum of infected necrosis with an increase in the incidence of gram-positive infection. PMID- 11865378 TI - Prospective evaluation of laparoscopy-assisted colectomy versus laparotomy with resection for management of complex polyps of the sigmoid colon. AB - Laparoscopy-assisted colectomy is technically feasible, but objective evidence of its benefits remains scarce. This study was done to evaluate the outcomes and operative stress of laparoscopy-assisted colectomy versus the traditional open method in the management of sigmoid complex polyps that cannot be safely or adequately removed by colonofibroscopy. Between January 1997 and December 1999, a total of 42 patients were equally randomized to the laparoscopy group and the laparotomy group by the blocked randomization method. Three patients randomized to the laparoscopy group did not complete the trial; therefore 18 patients treated by laparoscopy-assisted sigmoidectomy and the other 21 treated by the open method were prospectively evaluated. These two groups of patients were well matched in age, gender, symptoms, tumor location, localization method, tumor size, morphology, histopathology, and the accuracy of the clinical diagnosis. Two standardized surgical strategies, the lateral-to-medial and medial-to-lateral dissection sequences, were performed in 14 and 4 patients of the laparoscopy group, respectively, according to whether their tumors were located above or below 20 cm above the anal verge. After evaluating the surgical outcomes, we found that the laparoscopy group was significantly better than the laparotomy group in regard to parameters that included severity of postoperative pain, wound size, postoperative complication rate, and the duration of postoperative ileus, hospitalization, and disability. There was no significant difference in the operating times for these two groups. However, the costs of the laparoscopy group were significantly higher. To evaluate the surgical stress, we measured the serum C-reactive protein (CRP) level, erythrocyte sedimentation rate (ESR), total lymphocyte count, and CD4+/CD8+ ratio 24 hours before and after surgery. We found that the postoperative serum CRP level and the ESR were significantly less elevated and the total lymphocyte counts and CD4+/CD8+ ratio were significantly less depressed in the laparoscopy group than in the laparotomy group. We thus concluded that laparoscopy-assisted sigmoidectomy can be safely performed with shorter convalescence and less operative stress but at a higher cost. We strongly recommended the use of this technique in the management of sigmoid complex polyps if the patient's economic status permits. PMID- 11865379 TI - Lymph node recovery from colorectal tumor specimens: recommendation for a minimum number of lymph nodes to be examined. AB - Lymph node involvement is the most important prognostic factor for patients who have undergone radical surgery for colorectal carcinoma. An accurate examination of the surgical specimens is mandatory for the correct assessment of the lymph node status of the tumor. The risk of understaging is particularly high for patients with tumors classified as Dukes B (TNM stage II). The aim of this study was to determine if a specified minimum number of lymph nodes examined per surgical specimen could have any effect on the prognosis of patients who had undergone radical surgery for Dukes B colorectal cancer. Between 1988 and 1995 a total of 140 patients underwent radical resection of Dukes B colorectal cancer by the same surgeon (C.C.). The relation between clinicopathologic variables and survival was estimated using the Kaplan-Meier method. The Cox proportional hazard regression model was used to identify the variables that can independently influence survival. A median of 12 lymph nodes (range 3-38) was examined per tumor specimen. The 5-year survival rate of Dukes B patients who had had eight or fewer lymph nodes examined after surgery was 54.9%, whereas the survival rate for those who had had nine or more lymph nodes examined was 79.9% (p < 0.001). Cox regression analysis identified the number of lymph nodes as the only independent prognostic factor (p = 0.01). Seventy patients with one to four metastatic lymph nodes (Dukes C patients) who had been operated on during the same period were included in the survival analysis for comparison. The 5-year survival rate of the Dukes B patients with eight or fewer lymph nodes examined was similar to that of the 70 Dukes C patients (54.9% and 51.8%, respectively). Examination of eight or fewer lymph nodes in Dukes B colorectal patients may be considered a high risk factor for missing positive lymph nodes in the surgical specimens. Our results suggest that harvesting and examining a minimum of nine lymph nodes per surgical specimen may be sufficient for reliable staging of lymph node-negative tumors. PMID- 11865380 TI - Immunohistochemical expression of P53 and oncogenes in ulcerative colitis associated colorectal carcinoma. AB - The role of the tumor suppressor gene p53 and proto-oncogenes mdm-2, waf-1,and bcl-2 in sporadic colorectal carcinoma (CRC) has been well investigated. However, little is known about the role of these genes in the development of ulcerative colitis-associated colorectal carcinoma (CAC). Colectomy specimens from patients with CAC, patients with ulcerative colitis (UC) and dysplasia, patients with long standing UC without carcinoma or dysplasia, and patients with CRC were investigated in comparison to normal colon (NC) specimens from patients with diverticulosis without histologic signs of inflammation. Immunohistochemistry was performed with antibodies against p53, mdm-2, waf-1, and bcl-2; and staining was evaluated semiquantitatively with an expression of more than 20% of tumor cell nuclei or epithelial cell nuclei in nontumor specimens considered "positive." Statistical analysis was performed using Fisher's exact test. In carcinomas, p53 was positive in 50% of CRC tissues and 60% of CAC tissues without statistical difference. Positive expression of p53 was found in most high-grade dysplasia but not in low-grade dysplasia (p < 0.01). Whereas mdm-2 and bcl-2 were only sporadically expressed, waf-1 was observed in most specimens, with a high prevalence in UC without carcinoma or dysplasia (11/15). NC specimens were always negative for all antibodies. Immunohistochemical expression of p53, mdm-2, waf-1, and bcl-2 is similar for CAC and CRC. The malignant potential of dysplasia in UC is partially confirmed by a high prevalence of p53 and waf-1 expression, suggesting that CAC may develop along pathways that are different from CRC. High expression of waf-1 in nonmalignant long-standing UC has to be proved over a long term course in its role as an independent cancer risk factor in UC patients. PMID- 11865381 TI - [Anesthesiologist and phylogenesis - what are the conclusions based on biology?]. PMID- 11865383 TI - [Phylogenetics of the immunosystem]. AB - The human immune system is composed of two closely cooperating entities: innate, "unspecific" immune defense on one hand and adaptive, "specific" immunity on the other. Innate immune defense mechanisms were already developed very early in the evolution of the animal kingdom. By contrast, adaptive immunity exclusively evolved in jawed vertebrates within a surprisingly short time span. New data especially from genome sequencing projects now allow first glimpses on those events that resulted in the formation of adaptive immunity. PMID- 11865384 TI - [Anesthesia in neuromuscular disorders. Part 1: introduction]. AB - Disorders of skeletal muscle and peripheral nervous system are collectively called neuromuscular disorders (NMD). Important for anesthesia is that these disorders show various symptoms and have a high risk during general anesthesia. Especially administration of succinylcholine and volatile anaesthetics may cause problems. Under special circumstances opioids, nondepolarising muscle relaxants and intravenous anaesthetics can interfere with this kind of disorder, too. Complications during and after anaesthesia may result in malignant hyperthermia, malignant hyperthermia-like reactions and primary or secondary changes relating to the underlying NMD. These include cardiac and respiratory problems, dysautonomia as well as hypothermia or hyperthermia. Thus the perioperative management must be determined individually to assure the best possible safety for each patient. Preoperative examination such as ECG, echocardiography, respiratory function test including arterial blood-gas analysis, x-ray of the thorax, neurological status, and extended serum chemistry (such as CK and myoglobin) needs to be done. For premedication no drugs suppressing respiratory function should be administered. Regional anesthesia should be used whenever possible, especially in patients with respiratory and cardiac problems. The dosage of all recommended drugs should be as low as possible. Volatile anaesthetics should not be administered in the majority of NMD and succinylcholine is contraindicated, with the exception of myasthenia gravis. Additionally to the usual intraoperative monitoring, the invasive measurement of blood pressure allows frequent blood-gas analysis. It is obligate to monitor neuromuscular function and body temperature. During recovery special attention should be paid to maintain normal body temperature and electrolytes and acid-base status. The discharge of the patient from the recovery area to the normal ward should be performed only after respiratory function is normalized. PMID- 11865385 TI - [Tumescent technique for local anesthesia]. AB - Tumescent local anesthesia was originally used in liposuction but is also carried out for other plastic, cosmetic, dermatological procedures and for surgery of the venous system, often in outpatients. For this purpose, large amounts of fluids containing diluted lidocaine or prilocaine and epinephrine are infused subcutaneously. In this review of the literature, this technique is assessed in view of potential anesthesiological complications such as intoxication with lidocaine, prilocaine, overdosage of epinephrine or overload with fluids. While originally a lidocaine dosage of 35 mg/kg b.w. was considered to be safe, dosages were then increased to 55 mg/kg b. w. and even 90 mg/kg b. w. without data showing the safety of such high doses. Published data of plasma concentrations were obtained from small numbers of patients, showing that the concentrations were below 5 microg/ml which is considered the nontoxic range. The maximum levels were observed after 4 - 12 hours, if epinephrine was used. In a few patients, however, the values had not yet begun to decrease at the end of the 23 hours observation period. Replacing lidocaine by prilocaine shifts the problem of toxicity to that of the formation of methemoglobine, which can reach levels of more than 10 %. Data about effects of high-dose epinephrine in the literature are sparse, but tachycardia, arrhythmias and hypertension remain a major concern. Although fluids are applied subcutaneously, an overload with fluids may occur. Cases of lung edema have been reported, however, hypovolemia caused by a loss of fluid into the third space cannot be excluded. Because of these possible complications, tumescent local anesthesia should be employed in outpatients with great care. Patients should be monitored during the procedure and for a sufficient period of time thereafter by adequately trained staff. Patients with cardiac or pulmonary risk factors should not undergo tumescent local anesthesia. PMID- 11865386 TI - [Clonidine vs. Midazolam for premedication - comparison of the anxiolytic effect by using the STAI-test]. AB - OBJECTIVE: Many investigations described a positive effect of clonidine for premedication (reduction of perioperative stress response, analgetic effect, anaesthetic-saving effect, prophylaxis of postoperative shivering). The most important objective in premedication is undisputed anxiolysis. The anxiolytic effect of clonidine is differently discussed. Therefore we investigated the anxiolytic effect of clonidine in comparison to midazolam. METHODS: 50 patients (ASA 1 - 3) were included in the present doubleblinded, prospective study. These patients got an oral premedication with 1 mg flunitrazepam in the evening before surgery and 5 microg/kg(-1) clonidine or 100 microg/kg(-1) midazolam 60 - 90 minutes before surgery. By using Spielbergers State-Trait Anxiety Inventory (STAI G X1) we registered the degree of state-anxiety before and after premedication. RESULTS: Both clonidine as well as midazolam showed a significant reduction of the state-anxiety over time (p < 0,001). Between the groups there were no differences in the STAI-scores at any time (p > 0,05). The degree of reduction in the state-anxiety were comparable between the groups. CONCLUSION: In addition to the higher up listed effects clonidine showed a anxiolytic effect, which is comparable to midazolam. Therefore clonidine is an interesting alternative to benzodiazepines, which are commonly used drugs for premedication. PMID- 11865387 TI - [The early axial traction of the cervical spine after anaesthesia with intubation and extreme reclination of the head]. AB - OBJECTIVE: Patients who have an operation under general anaesthesia with muscle relaxation often complain about neck pain if the head needs to be placed in extreme reclination to facilitate surgical access. Patients complain about vertigo, light muscle tenseness but also about severe joint blockages in the neck region. Due to this complication the standard practise in some hospitals is to refer these patients routinely to a physiotherapist postoperatively. This study investigated the influence of an axial traction - a treatment which can easily be learned by anaesthesiologists - on blockages of the cervical spine in those patients. METHODS: In two randomised groups (n = 15 each) of preoperative inconspicuous patients the following directions of motion were investigated: Ante and retroflexion of C0/1, side inclination C0/1, side nodding and side movement C2/3 to C6/T1, dorsal movement C5/Th2. The examinations took place at the preoperative anaesthetic round, shortly before extubation, two hours after extubation and the next day. Additionally the patients were asked about their discomfort. An axial traction of the cervical spine was performed in one group after extubation. The number of new blockages and the subjective discomfort of the patients was compared with the Chi-Square test. RESULTS: An axial traction of the cervical spine reduces the frequency and the intensity of symptoms significantly. CONCLUSION: It was investigated whether an axial traction of the cervical spine - a treatment that can easily be learned by anaesthesiologists - could improve patients' comfort. The study showed that an axial traction of the cervical spine immediately after extubation reduces the frequency and intensity of symptoms significantly. This treatment is highly effective, not very time consuming and, if done correctly, without any risk for the patient. By using this treatment routinely, additional expenses for physiotherapeutic interventions could be reduced. PMID- 11865388 TI - [Registration of material consumption in anesthesia using a data management system]. AB - BACKGROUND: The aim of this study was to investigate whether an Anesthesia Information Management System (AIMS) can provide reliable data on the consumption of single-use anesthetic material without necessitating an expensive and time consuming inventory. To this end, the number of selected anesthesia-related materials and the total costs in orthopedic theatres for which the department of anesthesia had been charged in the year 2000 were compared to the data calculated by the AIMS. METHODS: Anesthesia-related material is provided by a computer-based system of storage facilities (KLIMA II) in the Department of Anesthesiology and Intensive Care Medicine at the University Hospital Giessen. All costs arising in orthopedic theatres are exclusively charged to one single account. At the same time, the online-documentation software, NarkoData (IMESO GmbH, Huttenberg, Germany), collects all data on consumption of anesthetic single-use material. The total amount of peripheral (PVC) and central-venous catheters (CVC), urinary catheters (UC) and endotracheal tubes (ET) used in the year 2000 was ascertained by the AIMS and compared to the respective data accounted by the administration. RESULTS: In the year 2000, the number of patients treated in orthopedic theatres totaled 1,865. By means of the AIMS, a consumption of 783 CVCs, 644 UCs and 949 ETs could be documented. In contrast, hospital administration had billed 880 CVCs, 700 UCs, and 1,050 ETs: discrepancies of 11.0 % for CVCs, 8.0 % for UCs and 9.6 % for ETs. Concerning the two most frequently used CVCs, the AIMS failed to document costs of 3,238 DM. For PVCs (16 gauge and 14 gauge), the official cost was 10.8 % and 46.7 % higher compared to the number documented by the AIMS. Since the number of PVCs totaled 3,400, the AIMS failed to document costs of 1,900 DM. CONCLUSION: Comparison of both methods revealed substantial deficits in documenting cost-relevant materials. There were no detailed data available on the whereabouts of the materials used, i.e. whether tubes and catheters were undocumented, used or discarded. However, the AIMS may provide additional valuable information about possible sources of material wastefulness. This is especially true for infrequently used anesthesia-related materials. PMID- 11865389 TI - [Anesthesiological management in patients with sickle cell disease]. AB - We report on the general anesthesia in the case of an orthopedic surgery conducted on two brothers with sickle cell anemia. After concerted planning with a pediatric hematologist concerning the perioperative management an exchange transfusion was conducted ambulatorily before surgery. The following anesthesia under continuous fluid infusion, warming and regular hemoglobin and temperature control could be conducted without problems. One of the boys was treated with two heterologous red cell packs after an unexpected hemoglobin drop intraoperatively. The postoperative course was uneventful, especially concerning sickle cell relevant complications. The case reports are intended to show up the pathogenesis of patient-relevant sickle cell crises and critically evaluate central points of anesthesiological management, especially the choice of preoperative transfusion. PMID- 11865390 TI - [Caesarean section after severe trauma and cardiac arrest - a case report]. AB - A 40 year-old woman with a prominent abdomen was hit by a car on the motorway. The emergency call was "cardiac arrest after multiple trauma in a woman in the late stages of pregnancy". The patient was asystole as indicated by the electrocardiogram 6 minutes after the primary emergency call. Cardiopulmonary resuscitation and endotracheal intubation was performed and a total dose of 3 mg epinephrine was given intravenously. The patient remained asystole. In this preclinical situation the suspected pregnancy could neither be excluded nor confirmed. At least 30 minutes would have been necessary to transfer the patient to the next hospital to perform a caesarean section under controlled conditions. A second physician - trained in obstetrics - was called to perform caesarean section preclinically. In addition, a special ambulance was called to provide sufficient postdelivery care of the neonate. However, the second physician who arrived 18 minutes later was not an obstetrician but also an anaesthesiologist who was trained in neonatal intensive care. At this time - 33 minutes after the first emergency call - the emergency physicians decided to perform a caesarean section immediately. However, the patient was not pregnant. Obviously, the primary emergency call was misleading. The skills of the emergency physicians and the conditions at the site of the emergency have to be weighed carefully against the expected time that is needed to reach the next hospital where a caesarean section can be performed under controlled conditions. In this case, both emergency physicians were anaesthesiologists and they had no doubt that the patient was in the late stages of pregnancy. Due to the prolonged cardiac arrest that did not respond to cardiopulmonary resuscitation both physicians were convinced that immediate preclinical caesarean section represented the only therapeutical option for the unborn patient. PMID- 11865391 TI - The role of donor CD4(+) T cells in the reconstitution of oral immunity by herpes simplex virus type 1 in severe combined immunodeficiency mice. AB - Severe combined immunodeficiency (SCID) mice with ill-developed Peyer's patches develop neither antibodies nor protection against lethal herpes simplex virus type 1 (HSV-1) infection by oral immunization. However, SCID mice carrying spleen cells from immunocompetent BALB/c mice had serum anti--HSV-1 antibody; anti--HSV 1 IgA antibody was detected in eye wash samples, and the mice were protected against lethal HSV-1 infection (88% survival rate). Western blotting showed that antibodies in SCID mice carrying spleen cells from BALB/c mice recognized 60-kDa HSV-1. The effector cells in transferred spleen cells were CD4(+), not CD8(+), T cells. Donor T cells were detected in the submucosal layer of the gut in SCID mice 1 day after transfer. Rapid movement of donor T cells to the gut may have a role in mucosal immunity to HSV-1. Thus, the normal environment for mucosal immunity develops in SCID mice without prior presence of CD4(+) T cells. PMID- 11865392 TI - The origin of hepatitis C virus reinfecting transplanted livers: serum-derived versus peripheral blood mononuclear cell-derived virus. AB - When hepatitis C virus (HCV) infection recurs after liver transplantation, it is unclear whether the liver graft is colonized by virions present in the circulation or by those associated with peripheral blood mononuclear cells (PBMC). In 6 HCV-infected transplant recipients, HCV sequences were analyzed by the single-strand conformational polymorphism (SSCP) assay and direct sequencing in pretransplant-paired PBMC and serum samples and in posttransplant follow-up serum samples. In 2 patients, SSCP patterns for pretransplant PBMC-serum pairs were identical, while in 4 patients they were different. In 3 patients from the latter group, the posttransplant viral sequences resembled those found in pretransplant serum samples, whereas in the other patient from that group, viral sequences after transplantation were transiently identical to those found in pretransplant PBMC. In HCV-positive liver transplant recipients, the liver graft is colonized primarily by liver-derived virus remaining in the circulation. However, virus variants of likely extrahepatic origin can be detected in serum early after transplantation. PMID- 11865393 TI - Risk factors and outcome of varicella-zoster virus pneumonia in pregnant women. AB - To determine the factors associated with an increased risk of developing varicella-zoster virus (VZV) pneumonia during pregnancy, a case-control analysis was done in which 18 pregnant women with VZV pneumonia were compared with 72 matched control subjects. VZV infection was identified clinically, and VZV pneumonia was diagnosed by dyspnea and findings on chest radiographs. Of 347 pregnant women with VZV infection, 18 (5.2%) had pneumonia treated with acyclovir, and none died. Mean gestational age at rash onset was 25.8 plus minus 8.8 weeks for patients with pneumonia and 17.7 +/- 10.3 weeks for control subjects, which was not significant in the multivariable model. Women with VZV pneumonia were significantly more likely to be current smokers (odds ratio [OR], 5.1; 95% confidence interval [CI], 1.6-16.7) and to have > or = 100 skin lesions (OR, 15.9; 95% CI, 1.9-130.2). Pregnant women with VZV infection may be more likely to develop varicella pneumonia if they are smokers or manifest > or = 100 skin lesions. PMID- 11865395 TI - Antibody-dependent cell-mediated cytotoxicity in cervical lavage fluids of human immunodeficiency virus type 1--infected women. AB - Studies were designed to determine whether cervical antibodies in human immunodeficiency virus type 1 (HIV-1)-infected women participate in antibody dependent cell-mediated cytotoxicity (ADCC). Serum and cervical lavage fluid (CVL) ADCC titers were compared with plasma virus load and CD4 cell number in 45 infected and 10 uninfected women from the Women's Interagency HIV Study. Serum and CVL were incubated with normal peripheral blood lymphocytes and HIV-1 gp120 bearing target cells in a standard (51)Cr-release assay. When stringent criteria were used to define ADCC activity, 63% had activity in > or = 1 fluid sample, 56% had serum titers, and 16% had CVL titers. Serum titers did not predict CVL titers. Three women with CVL ADCC had no serum ADCC, which suggests that ADCC antibodies may be produced locally. ADCC antibodies are present in the cervicovaginal fluids, which indicates that this form of innate immunity can contribute to mucosal defense against HIV-1. PMID- 11865394 TI - Correlates of nontransmission in US women at high risk of human immunodeficiency virus type 1 infection through sexual exposure. AB - Seventeen women who were persistently uninfected by human immunodeficiency virus type 1 (HIV-1), despite repeated sexual exposure, and 12 of their HIV-positive male partners were studied for antiviral correlates of non-transmission. Thirteen women had > or = 1 immune response in the form of CD8 cell noncytotoxic HIV-1 suppressive activity, proliferative CD4 cell response to HIV antigens, CD8 cell production of macrophage inflammatory protein-1 beta, or ELISPOT assay for HIV-1 specific interferon-gamma secretion. The male HIV-positive partners without AIDS had extremely high CD8 cell counts. All 8 male partners evaluated showed CD8 cell related cytotoxic HIV suppressive activity. Reduced CD4 cell susceptibility to infection, neutralizing antibody, single-cell cytokine production, and local antibody in the women played no apparent protective role. These observations suggest that the primary protective factor is CD8 cell activity in both the HIV positive donor and the HIV-negative partner. These findings have substantial implications for vaccine development. PMID- 11865396 TI - T69D/N pol mutation, human immunodeficiency virus type 1 RNA levels, and syncytium-inducing phenotype are associated with CD4 cell depletion during didanosine therapy. AB - The contribution of virologic and host factors to CD4 cell depletion associated with human immunodeficiency virus (HIV) type 1 was evaluated in children drawn from a larger efficacy trial of 2 doses of didanosine (ddI) monotherapy (Pediatric AIDS Clinical Trials Group 144). Thirty children, half with stable CD4 cell counts (non-progressors) and half with a marked decline in CD4 cells (progressors), were studied during 60-72 weeks of ddI therapy. The children were matched for age and CD4 cell counts at study entry. Three viral parameters, syncytium-inducing phenotype, higher virus load, and mutation in HIV-1 pol encoding the T69D/N mutation, were associated with disease progression. Disease progression was not associated with mutations in the reverse-transcriptase gene previously associated with resistance to ddI (L74V, K65R, or M184V). The selection of the T69D/N mutation in children with HIV-1 disease progression during ddI therapy suggests that this mutation confers a fitness advantage to the virus that may include resistance to ddI. PMID- 11865397 TI - Antiretroviral therapy reduces markers of endothelial and coagulation activation in patients infected with human immunodeficiency virus type 1. AB - We investigated the effect of antiretroviral therapy on vascular activation in 41 human immunodeficiency (HIV)--infected patients receiving a regimen that included either at least 1 protease inhibitor (PI; n = 21) or a nonnucleoside reverse transcriptase inhibitor (NNRTI; n = 20). A control group of 21 healthy subjects was included for comparison. Levels of endothelial markers (soluble vascular cell adhesion molecule [sVCAM]--1, soluble intercellular adhesion molecule--1, and von Willebrand factor) were higher in HIV-infected persons before treatment than in control subjects and decreased significantly after 5--13 months of treatment. Levels of sVCAM-1 and von Willebrand factor correlated significantly with initial virus load. d-dimer concentrations also decreased significantly after initiation of treatment. PI- and NNRTI-containing regimens had similar effects. Therapy did not reduce levels of the soluble platelet (sP) activation markers sP-selectin and CD40 ligand. The inhibition of markers of vascular activation may counterbalance sequelae of therapy-induced dyslipidemia and potentially prevent development of atherosclerosis in HIV-infected patients. PMID- 11865398 TI - Early CD4(+) T cell recovery in human immunodeficiency virus-infected patients receiving effective therapy is related to a down-regulation of apoptosis and not to proliferation. AB - This prospective study investigated the contributions of apoptosis and proliferation of CD4(+) T cells obtained by the introduction of a new antiretroviral treatment for human immunodeficiency virus infection. Virus load; T cell counts; apoptosis of T cell subsets, including naive cells; and proliferation were determined from treatment initiation to the third month in a cohort of patients. An increase in CD4(+) T cell count > or = 100 cells/microL over baseline was considered to be a satisfactory immune reconstitution. Sixty nine patients completed the protocol, 22 of whom met our definition of a satisfactory immune reconstitution, showing a significantly more pronounced reduction in spontaneous CD4(+) T cell apoptosis at month 1 as well as month 3, compared with the other patients. In contrast, neither Fas-induced apoptosis down regulation nor Fas-induced increased proliferation capacity was associated with a satisfactory immune reconstitution. Down-regulation of CD4(+) T cell apoptosis by antiretroviral treatment is the main mechanism associated with early CD4(+) T cell increase. PMID- 11865399 TI - Predictors of long-term increase in CD4(+) cell counts in human immunodeficiency virus-infected patients receiving a protease inhibitor-containing antiretroviral regimen. AB - The temporal relationships between plasma human immunodeficiency virus (HIV) RNA levels and evolution of CD4(+) cell counts was studied, using a 2-slope longitudinal mixed model, in 988 patients prospectively enrolled at the initiation of a protease inhibitor--containing regimen of antiretroviral therapy. The short-term slope (baseline through month 4) for mean change in CD4(+) cell count was +21.2 cells/mm(3)/month, and the long-term slope (month 4 through month 24) was +5.5 cells/mm(3)/month. Compared with results from patients without viral response, the long-term slope was 2.5 cells/mm(3)/month higher in patients who had plasma HIV RNA levels of <500 copies/mL at month 4 (P <.001). It was significantly lower after a rebound in plasma HIV RNA level to > or = 500 copies/mL (P <.0001), varied according to plasma HIV RNA level at the time of rebound, and was negative only when the plasma HIV RNA level at rebound was > or = 10,000 copies/mL. If CD4(+) cell counts can remain elevated despite virologic treatment failure, such a discrepant response may be transient in patients who have a high plasma HIV RNA level at the time of treatment failure. PMID- 11865400 TI - Risk factors for the transmission of methicillin-resistant Staphylococcus aureus in an adult intensive care unit: fitting a model to the data. AB - Little is known about the amount of cross-transmission, the risk factors for infection, and the relative effectiveness of infection control procedures when methicillin-resistant Staphylococcus aureus (MRSA) infection occurs at highly endemic levels in intensive care units. A cohort study was done to identify exposures associated with cases that likely were the result of cross-transmission (i.e., occurring in clusters and with indistinguishable MRSA macrorestriction profiles). Fitting a simple stochastic model to the ascertained data allowed prediction of the effectiveness of infection control measures. Exposure to relative staff deficit (adjusted rate ratio, 1.05 independent; 95% confidence interval, 1.02-1.09) was the only factor significantly associated with potential transmission (P =.001). It was predicted that a 12% improvement in adherence to hand-hygiene policies might have compensated for staff shortage and prevented transmission during periods of overcrowding, shared care, and high workload but that this would be hard to achieve. PMID- 11865401 TI - Inhibition of pneumococcal carriage in mice by subcutaneous immunization with peptides from the common surface protein pneumococcal surface adhesin a. AB - Pneumococcal surface adhesin A (PsaA), a common protein expressed on all 90 pneumococcal serotypes, is a vaccine candidate. Three anti-PsaA monoclonal antibody phage display-expressed monopeptides (15 mers), in various formulations as lipidated or nonlipidated multiantigenic peptides or as bi- or tripeptide constructs, were studied in a mouse nasopharyngeal carriage model to determine the inhibitory effect of induced antibodies on carriage of pneumococcal serotypes 2, 4, and 6B. Antibodies to each of the various peptides tested reduced carriage of the 3 serotypes. Reduction in carriage by nonlipidated multiantigenic peptide antibodies was highly variable (39%-94%; mean, 59%; standard deviation [SD], 20.2%); however, more-consistent results were observed in mice immunized with lipidated (56%-98%; mean, 69%; SD, 13.6%) and combination or bipeptide (55%-91%; mean, 76%; SD, 13.1%) formulations. These peptides are immunogenic, and their induced antibodies reduce carriage in mice. PsaA peptides demonstrate potential for being important new vaccines against pneumococcal carriage, otitis media, and invasive pneumococcal disease. PMID- 11865402 TI - Prevalence of enteric pathogens among international travelers with diarrhea acquired in Kenya (Mombasa), India (Goa), or Jamaica (Montego Bay). AB - Stools from tourists from Europe and North America who acquired diarrhea in Mombasa (Kenya), Goa (India), or Montego Bay (Jamaica) were examined for enteric pathogens. Enterotoxigenic Escherichia coli (ETEC) was the most common pathogen (25%) identified in the 3 locations. Isolation of Shigella species was more frequent in Goa and Mombasa than in Montego Bay (10%, 9%, and 0.3%, respectively; P <.005). Viruses (rotaviruses and enteric adenoviruses) were found in 9% of travelers to the 3 areas. Of 275 ETEC isolates in this study, 158 (57%) produced a defined colonization factor antigen (CFA). Coli surface 6 (CS6) was the most frequent and was found in 41%-52% of CFA/CS-positive ETEC isolates. The frequency of resistance among bacterial enteropathogens to traditional antimicrobial agents was particularly high throughout the study period in all 3 regions. Quinolones were active against the bacterial enteropathogens in the 3 sites. PMID- 11865403 TI - Lewis antigen expression and other pathogenic factors in the presence of atrophic chronic gastritis in a European population. AB - To study the relationship between Helicobacter pylori cagA and vacA status and the expression of Lewis (Le) antigens and between these characteristics and atrophic chronic gastritis (ACG), H. pylori infection was assessed by culture and by histologic and serologic tests, cagA and vacA were assessed by a polymerase chain reaction--based reverse hybridization assay, and bacterial Le expression was assessed by immunoblotting. ACG was any form of antral or fundic atrophy with or without intestinal metaplasia. Of the 215 isolates, 64% were cagA(+) and 100% were vacA(+) (s1m1, 42%; s1m2, 29%; s2m2, 29%; and s2m1, 0). Le typing of 155 isolates showed that 6 (4%) were Le(x), 31 (20%) were Le(y), 87 (56%) were Le(x,y), and 31 (20%) were neither Le(x) nor Le(y). Two main clusters of isolates were identified by multiple correspondence analysis: s1a/m1/cagA(+)/Le(x)+/Le(y)+ (n=44; 29.7%) and s2/m2a/cagA(-)/Le(y)+ or Le(x)-/Le(y)- (n=29; 19.7%). Among patients with ACG, 54% of their isolates were from cluster s1m1/cagA(+)/Le(x)+/Le(y)+, which was associated with the presence of ACG (odds ratio, 7.4; 95% confidence interval, 1.5-37.0). PMID- 11865404 TI - Differential T and B cell responses against Mycobacterium tuberculosis heparin binding hemagglutinin adhesin in infected healthy individuals and patients with tuberculosis. AB - Because only 10% of individuals infected with Mycobacterium tuberculosis will eventually develop disease, antigens that are recognized differently by the immune systems of infected healthy and diseased subjects may constitute potential vaccine candidates. Here, the heparin-binding hemagglutinin adhesin (HBHA) is identified as such an antigen. Lymphocytes from 60% of healthy infected individuals (n=25) produced interferon (IFN)-gamma after stimulation with HBHA, compared with only 4% of patients with active tuberculosis (n=24). In the responders, both CD4(+) and CD8(+) cells secreted HBHA-specific IFN-gamma, and the antigen was presented by both major histocompatibility complex class I and II molecules. In contrast to the reduced ability of patients with tuberculosis to produce HBHA-specific IFN-gamma, most of them (82%) produced anti-HBHA antibodies, compared with 36% of the infected healthy subjects. These observations indicate that HBHA is recognized differently by the immune systems of patients with tuberculosis and infected healthy individuals and might provide a marker for protection against tuberculosis. PMID- 11865405 TI - Essential role of the Candida albicans transglutaminase substrate, hyphal wall protein 1, in lethal oroesophageal candidiasis in immunodeficient mice. AB - Oroesophageal candidiasis is caused by the combined action of fungal virulence factors and host inflammatory responses when protective immunity is absent. Hyphal wall protein 1 (Hwp1) on germ tubes and true hyphae of Candida albicans forms covalent cross-links to buccal epithelial cells in vitro by functioning as a substrate for mammalian transglutaminases. In this study, beige-athymic (bg/bg nu/nu) or transgenic epsilon 26 mice that have combined natural killer and T cell defects did not succumb to candidiasis after oral administration of C. albicans strains with inactivated HWP1 genes, whereas mice given isogenic strains of C. albicans that had a single copy of HWP1 survived only 2-3 weeks. Illness correlated with extensive alterations of the lingual and esophageal mucosa that were absent in mice given the hwp1/hwp1 mutant. HWP1 is a promising target for development of antifungal drugs for treatment of oroesophageal candidiasis. PMID- 11865406 TI - Triggering of innate immune responses by schistosome egg glycolipids and their carbohydrate epitope GalNAc beta 1-4(Fuc alpha 1-2Fuc alpha 1-3)GlcNAc. AB - To investigate the interactions of glycoconjugates with the innate immune system, peripheral blood mononuclear cells were stimulated with glycolipids derived from Schistosoma mansoni eggs and worms and with biochemically synthesized neoglycoconjugates. Egg glycolipids stimulated the production of interleukin (IL) -10, IL-6, and tumor necrosis factor--alpha in monocytes, whereas worm glycolipids failed to do so. When monoclonal antibodies that specifically recognize defined carbohydrate epitopes were used, the binding of a GalNAc beta 1 4(Fuc alpha 1-2Fuc alpha 1-3)GlcNAc (LDN-DF) reactive antibody was pronounced on egg glycolipids but was absent on worm glycolipids. The binding of antibodies that recognize Gal beta 1-4(Fuc alpha 1-3)GlcNAc (LewisX), GalNAc beta 1-4GlcNAc (LDN), and GalNAc beta 1-4(Fuc alpha 1-3)GlcNAc (LDN-F) was comparable for both preparations. Cytokine production in response to neoglycoconjugates containing enzymatically synthesized glycans also was measured. The LDN-DF neoglycoconjugate was the most potent cytokine inducer, which indicates that this difucosylated glycan can act at the host-parasite interface and can trigger innate immune responses. PMID- 11865408 TI - Persons with early syphilis identified through blood or plasma donation screening in the United States. AB - The number of persons with early syphilis who donated blood between 1995 and 2000 in the United States was estimated using data collected in the National Electronic Telecommunication System for Surveillance (NETSS). To distinguish paid from volunteer donors, cases reported in 2000 were analyzed. For the 6 years, 22 primary, 81 secondary, and 413 early latent syphilis cases were identified through donation screening. In 2000, 69 cases of early syphilis were identified through donation screening in 16 states. In 6 states that reported 53 of these cases, 31 case subjects (58%) were volunteer donors and 22 (42%) were paid donors. Eighty-one percent of volunteer donors and 64% of paid donors reported no risk factors for syphilis. After adjustment for variation in NETSS use, it was estimated that, over the 6 years, approximately 1200 cases of early syphilis were detected nationally through donation screening, and 58% of the case subjects were volunteer donors. PMID- 11865407 TI - Rhinovirus infection increases 5-lipoxygenase and cyclooxygenase-2 in bronchial biopsy specimens from nonatopic subjects. AB - Rhinovirus infections cause wheeze, cough, and bronchial hyperresponsiveness. To investigate the involvement of cysteinyl-leukotrienes and prostanoids in these symptoms, bronchial biopsy specimens from 9 normal subjects (nonatopic and with no history of chronic lung disease) were immunostained for 5-lipoxygenase (5-LO) and cyclooxygenase (COX) pathway enzymes 2 weeks before and 4 days after experimental infection with human rhinovirus serotype 16. 5-LO-positive cell counts increased 9-fold (from 0.48 to 4.4 cells/mm(2); P <.05), and 5-LO activating protein (FLAP)-positive cell counts increased 3.6-fold (from 1.8 to 6.5 cells/mm(2); P =.09). Levels of leukotriene A(4) hydrolase and leukotriene C(4) synthase were unchanged. COX-2--positive cell counts increased from 0 to 2.6 cells/mm(2) (P =.009), with no change in COX-1 levels. Increases of 3-4-fold were seen in levels of macrophages (P =.02) and mast cells (P =.07) but not of eosinophils (P >.4), and bronchoalveolar lavage fluid cysteinyl-leukotriene levels doubled (from 11.2 to 20.4 pg/mL; P =.13). Cold symptom scores correlated with bronchial immunostaining for FLAP (rho = 0.93; P =.001). In normal subjects, rhinovirus colds induce bronchial inflammation with markedly enhanced expression of 5-LO pathway proteins and COX-2. PMID- 11865409 TI - Ferrets as a model system for renal disease secondary to intestinal infection with Escherichia coli O157:H7 and other Shiga toxin-producing E. coli. AB - Ferrets were evaluated as a possible small animal model for the development of colitis and/or signs of the hemolytic uremic syndrome after oral infection with Escherichia coli O157:H7 or other Shiga toxin--producing E. coli (STEC). Ferrets treated with streptomycin (Stm) had higher counts of E. coli O157:H7 strain 86-24 Stm-resistant (Stm(r)) or O91:H21 strain B2F1 Stm(r) in their stools than non- Stm-treated animals. None of the animals displayed evidence of colitis, but Stm treated animals fed strain 86-24 Stm(r) exhibited weight loss significantly greater than that exhibited by ferrets fed an isogenic mutant negative for the adhesin intimin. Moreover, 11 (23%) of the 47 Stm-treated ferrets inoculated with 86-24 Stm(r) or B2F1 Stm(r) developed hematuria and/or histological damage to glomeruli or thrombocytopenia, compared with 0 of 14 uninfected control animals receiving Stm in water. Thus, the ferret may serve as a model for renal disease secondary to intestinal infection with STEC. PMID- 11865411 TI - Restricting the selection of antibiotic-resistant mutant bacteria: measurement and potential use of the mutant selection window. AB - The selection of antibiotic-resistant mutant bacteria is proposed to occur in a drug concentration range (the mutant selection window) that extends from the minimum inhibitory concentration (MIC) of susceptible cells to the MIC of the least susceptible, single-step bacterial mutants (the mutant prevention concentration [MPC]). MPCs were estimated for tobramycin, chloramphenicol, rifampicin, penicillin, vancomycin, and several fluoroquinolones by use of Escherichia coli and Staphylococcus aureus. Comparisons among reported serum drug levels indicate that new fluoroquinolones are the least likely to enrich populations of resistant mutant bacteria during monotherapy. These data partly explain the selective enrichment of populations of resistant mutant bacteria in medical practice. The mutant selection window range (MPC:MIC) was narrowed for fluoroquinolones by structure modification, pointing to a new direction in antibiotic refinement. The mutant selection window and the MPC were determined for combinations of rifampicin and tobramycin, using S. aureus, as a guide for combination therapy with compounds that alone cannot block enrichment of mutant bacterial populations. PMID- 11865410 TI - Temporal sequence and functional implications of V beta-specific T cell receptor down-regulation and costimulatory molecule expression following in vitro stimulation with the staphylococcal superantigen Toxic shock syndrome toxin-1. AB - The superantigen toxic shock syndrome toxin-1 (TSST-1) is implicated as the major cause of staphylococcal toxic shock syndrome. The temporal sequence of early signaling events in human peripheral blood mononuclear cells following TSST-1 stimulation was examined. TSST-1 induced rapid and complete down-regulation of V beta 2-specific T cell receptor (TCR), followed by transient CD154 expression on CD4(+) lymphocytes. This was sequentially followed by the up-regulation of CD86, CD80, CD40, and human leukocyte antigen-DR expression on CD14(+) monocytes. In contrast, S14N, a TSST-1 mutant toxin with a single amino acid substitution that is known to be impaired in interleukin (IL)--2, interferon (IFN)-gamma, and tumor necrosis factor (TNF)-alpha secretion, was deficient in both V beta 2-TCR down regulation and CD154 and CD80/CD86 expression. Furthermore, pretreatment with monoclonal antibodies against V beta 2-TCR, CD80/CD86, and CD154 significantly inhibited TSST-1-induced IL-2, IFN-gamma, and TNF-alpha secretion. Taken together, these results indicate that early V beta-specific TCR activation, along with CD80/CD86 and CD154 costimulation, are key determinants of the TSST-1 induced proinflammatory cytokine response. PMID- 11865412 TI - Maternal drug use is a preeminent risk factor for mother-to-child hepatitis C virus transmission: results from a multicenter study of 1372 mother-infant pairs. AB - This prospective multicenter study evaluated separately the significance of maternal injection drug use (IDU) and human immunodeficiency virus type 1 (HIV-1) coinfection in vertical transmission of hepatitis C virus (HCV). In all, 1372 consecutive, unselected HCV antibody-positive mothers and their infants were studied. Maternal HIV-1 coinfection (crude odds ratios [OR], 1.41; 95% confidence interval [CI], 1.16-1.66; P =.007) and IDU (OR, 1.58; 95% CI, 1.37-1.78; P <.00001) were linked to mother-to-child HCV transmission in unadjusted analysis when all anti-HCV-positive mothers were evaluated. When only HCV RNA-positive mothers were evaluated, maternal IDU, but not maternal HIV-1 coinfection, was significantly associated with mother-to-child HCV transmission. Multivariable analysis confirmed the link between maternal IDU and HCV transmission (adjusted OR [AOR], 1.51; 95% CI, 1.19-1.92; P =.0006), but no association was found with HIV-1 coinfection (AOR, 0.98; 95% CI, 0.73-1.33; P =.93). IDU, but not HIV-1 coinfection, seems to be a preeminent risk factor for vertical HCV transmission. PMID- 11865414 TI - Human cytomegalovirus enhances chemokine production by lipopolysaccharide stimulated lamina propria macrophages. AB - To elucidate the role of mucosal macrophages in intestinal human cytomegalovirus (HCMV) disease, primary lamina propria macrophages (LPM) were isolated from normal human jejunum, infected with HCMV, and studied for their cytokine responses. HCMV infection of LPM was confirmed by the presence of HCMV IE72 (UL123), pp65 (UL83), and glycoprotein B (UL55) proteins, which were detected by immunofluorescence, beginning at postinfection (pi) day 3, and were sustained through pi day 12 in 0.1%-0.5% of LPM. The late protein pp28 (UL99) was also detected up to pi day 12, consistent with productive infection. HCMV infection in LPM was characterized by quantitative competitive polymerase chain reaction, with maximum levels of HCMV DNA detected at pi day 7. HCMV infection of the LPM augmented lipopolysaccharide-inducible chemokine (interleukin [IL]-8 and macrophage inflammatory protein-1alpha) and cytokine (IL-6) production. These findings suggest that mucosal macrophages, via enhanced mediator production, play an important role in intestinal inflammation associated with HCMV infection. PMID- 11865413 TI - Variability of the nonstructural 5A protein of hepatitis C virus type 3a isolates and relation to interferon sensitivity. AB - The nonstructural 5A (NS5A) protein of hepatitis C virus (HCV) genotype 1 is thought to interact with several cellular proteins, including the double-stranded RNA-dependent protein kinase (PKR) induced by interferon (IFN). The PKR-binding domain (PKR-BD; aa 2209-2274), including the IFN sensitivity-determining region (aa 2209-2248) and other regions, could be linked to IFN resistance. Thus, the entire NS5A sequence of 27 isolates of HCV genotype 3a was investigated in relation to the clinical response to IFN. The NS5A 3a protein presented a low variability with some specific variable regions. Differential analysis between IFN-resistant and -sensitive isolates identified 5 regions in NS5A, 2 of them inside the PKR-BD and another around the variable 3 region. However, using the yeast growth suppression assay, no interaction was found between 5 resistant NS5A 3a proteins and PKR. Some amino acid changes of the NS5A protein of genotype 3a seemed to relate to IFN resistance independently of the PKR pathway. PMID- 11865415 TI - Accumulation of defective neuraminidase (NA) genes by influenza A viruses in the presence of NA inhibitors as a marker of reduced dependence on NA. AB - With the use of neuraminidase (NA) inhibitors (BCX-1812, oseltamivir, or zanamivir), drug-resistant variants of influenza A viruses were generated that lacked characteristic markers of resistance, such as substitutions in the NA active center or in the hemagglutinin. Drug resistance was associated with the accumulation of defective (Delta) RNA segments encoding NA. This phenomenon could be explained by reduced dependence of the virus on its NA activity. Analysis of the last isolates recovered from 11 volunteers, experimentally infected with influenza virus and treated with BCX-1812, revealed that they maintained full susceptibility to the drug in the NA inhibition assay (50% inhibitory concentration, 0.35-0.5 nM). The presence of DeltaRNA segments was detected in 1 of these isolates but was not found in the isolates recovered from placebo recipients (n = 8). Because of a lack of cell culture-based assays for susceptibility testing of human influenza viruses, detection of DeltaRNA segments should be considered an additional assay for monitoring of NA inhibitor resistance. PMID- 11865417 TI - Emergence of autologous neutralization-resistant variants from preexisting human immunodeficiency virus (HIV) quasi species during virus rebound in HIV type 1 infected patients undergoing highly active antiretroviral therapy. AB - The role of neutralizing antibodies (NAbs) during virus rebound in human immunodeficiency virus type 1 (HIV-1)-infected patients undergoing highly active antiretroviral therapy is poorly understood. Three patients in this study had NAbs to preexisting autologous HIV-1 and an episode of virus rebound after a prolonged period of virus suppression. To investigate the influence of NAbs on virus evolution, envelope genotypes of preexisting and rebound viruses were examined. Phylogenetic analysis of env (V1-V5) sequences indicated that rebound viruses had evolved from or preexisted in baseline populations. By use of envelope pseudotype viruses, rebound viruses were found to be significantly resistant to neutralization by autologous antibody in all 3 patients, indicating that rebound viruses were selected by NAbs. The site responsible for conferring neutralization resistance against autologous antibody was identified in the upstream C3 region in 2 of 3 patients. PMID- 11865416 TI - Safety and antiviral activity at 48 weeks of lopinavir/ritonavir plus nevirapine and 2 nucleoside reverse-transcriptase inhibitors in human immunodeficiency virus type 1-infected protease inhibitor-experienced patients. AB - The safety and antiviral activity of lopinavir (Lpv), a protease inhibitor (PI) coformulated with ritonavir (Rtv) to enhance its pharmacokinetic properties, were evaluated in 70 patients with plasma human immunodeficiency virus type 1 (HIV-1) RNA levels of 1000-100,000 copies/mL on a first PI-containing regimen. Patients were randomized to substitute only the PI with Lpv/Rtv, 400/100 mg or 400/200 mg twice daily. On day 15, nevirapine (200 mg 2x/day) was added, and nucleoside reverse-transcriptase inhibitors were changed. Despite a >4-fold reduction in phenotypic susceptibility to the preentry PI in 63% of patients, mean plasma HIV 1 RNA levels declined by 1.14 log(10) copies/mL after 2 weeks of Lpv/Rtv. At week 48, 86% of subjects receiving treatment had plasma HIV-1 RNA levels of <400 copies/mL; 76% had levels <50 HIV-1 RNA copies/mL (intent-to-treat: 70% and 60%, respectively). Mean CD4 cell counts increased by 125 cells/muL. Three patients discontinued therapy for drug-related adverse events. PMID- 11865418 TI - Prospective study of a serogroup X Neisseria meningitidis outbreak in northern Ghana. AB - After an epidemic of serogroup A meningococcal meningitis in northern Ghana, a gradual disappearance of the epidemic strain was observed in a series of five 6 month carriage surveys of 37 randomly selected households. As serogroup A Neisseria meningitidis carriage decreased, an epidemic of serogroup X meningococcal carriage occurred, which reached 18% (53/298) of the people sampled during the dry season of 2000, coinciding with an outbreak of serogroup X disease. These carriage patterns were unrelated to that of Neisseria lactamica. Multilocus sequence typing and pulsed-field gel electrophoresis of the serogroup X bacteria revealed strong similarity with other strains isolated in Africa during recent decades. Three closely related clusters with distinct patterns of spread were identified among the Ghanian isolates, and further microevolution occurred after they arrived in the district. The occurrence of serogroup X outbreaks argues for the inclusion of this serogroup into a multivalent conjugate vaccine against N. meningitidis. PMID- 11865419 TI - Sequence stability of the gene encoding outer membrane protein P2 of nontypeable Haemophilus influenzae in the human respiratory tract. AB - Nontypeable Haemophilus influenzae (NTHI) is an important cause of lower respiratory tract infections in patients with chronic obstructive pulmonary disease. Recent findings suggest that the major outer membrane protein P2 should be reconsidered as a vaccine candidate for NTHI. A P2-based vaccine would require a relative degree of sequence stability of the gene encoding P2 (ompP2) during colonization. To characterize the sequence stability of ompP2 during colonization of the human respiratory tract, ompP2 genes from 13 sets of isolates that persisted in patients with chronic obstructive pulmonary disease (mean colonization, 7 months) were sequenced. In 9 sets of isolates, ompP2 did not change. Sequence changes were noted in 4 sets of isolates. Most of these changes occurred within areas of repetitive DNA, suggesting that this type of DNA has a role in antigenic variation of P2. The sequence of ompP2 is relatively stable during persistence of NTHI in the human host. PMID- 11865420 TI - Systemic and mucosal antibody response to Moraxella catarrhalis after exacerbations of chronic obstructive pulmonary disease. AB - To characterize the immune response to Moraxella catarrhalis after exacerbations of chronic obstructive pulmonary disease (COPD), pre- and postexacerbation serum and sputum supernatant samples obtained during 21 exacerbations in 18 patients were studied, using the homologous infecting isolates. New serum immunoglobulin G (IgG) detected by whole-cell enzyme-linked immunosorbent assay developed after 12 (57.1%) of 21 exacerbations. Analysis of serum samples with flow cytometry, which detects antibodies that are exclusive to epitopes on the bacterial surface, revealed that 5 (23.8%) of the 21 exacerbations were associated with the development of new serum IgG to surface epitopes. Three of these serum samples and 2 other serum samples contained new IgG directed at lipooligosaccharide. Flow cytometry revealed that new mucosal IgA to surface-exposed epitopes of the infecting isolate developed in sputum supernatants after 42% of exacerbations. Therefore, adults with COPD develop variable humoral immune responses to M. catarrhalis after exacerbations, including new serum IgG and new mucosal IgA to epitopes on the bacterial surface. PMID- 11865421 TI - Identification and evolution of an IS6110 low-copy-number Mycobacterium tuberculosis cluster. AB - A cohort of 56 patients infected with related strains of Mycobacterium tuberculosis, the S75 group, was identified in a New Jersey population-based study of all isolates with a low number of copies of the insertion element IS6110. Genotyping was combined with surveillance data to identify the S75 group and to elucidate its recent evolution. The S75 group had similar demographic and geographic characteristics. Seventeen persons (30%) were linked epidemiologically. The S75 group was segregated from other low-copy-number isolates on the basis of several independent molecular methods. This group included 3 IS6110 genotype variants: BE, H6, and C28, containing 1, 2, and 3 IS6110 insertions, respectively. IS6110 insertion site mapping and comparative sequence analysis strongly suggest a stepwise acquisition of IS6110 elements from BE to H6 to C28. S75 represents a locally produced strain cluster that has recently evolved. The combination of multiple molecular tools with traditional epidemiology provides novel insights into dissemination, local transmission, and evolution of M. tuberculosis. PMID- 11865422 TI - The clinical significance of cerebrospinal fluid levels of kynurenine pathway metabolites and lactate in severe malaria. AB - A retrospective study of 261 Vietnamese adults with severe malaria was conducted to determine the relationship between cerebrospinal fluid (CSF) levels of metabolites of the kynurenine pathway, the incidence of neurologic complications, and the disease outcome. Three metabolites were measured: the excitotoxin quinolinic acid (QA); the protective receptor antagonist kynurenic acid (KA); and the proinflammatory mediator picolinic acid (PA). These measurements were related prospectively to CSF lactate levels. QA and PA levels were elevated, compared with those of controls. There was no difference in the levels of KA between these groups. Although >40% of malaria patients had QA CSF concentrations in the micromolar range, there was no association with convulsions or depth of coma. Levels of QA and PA were associated significantly with death, but a multivariate analysis suggested that these elevations were a consequence of impaired renal function. CSF lactate remained an independent and significant predictor of poor outcome. PMID- 11865423 TI - Merozoite surface protein 3 and protection against malaria in Aotus nancymai monkeys. AB - A blood-stage vaccine based on Plasmodium falciparum merozoite surface protein 3 (MSP3) was tested for efficacy in a primate model. Aotus nancymai monkeys were vaccinated with yeast-expressed MSP3 before a lethal challenge with Plasmodium falciparum parasites. Five of 7 control monkeys had acute infections and required treatment to control parasitemia. Only 1 of 7 monkeys vaccinated with MSP3 required this treatment. The efficacy of the MSP3 vaccination appeared to be comparable to that of MSP1(42), a leading asexual vaccine candidate, in response to which 2 monkeys experienced acute infections. In the MSP3-vaccinated group, protection correlated with prechallenge titers of antibody to MSP3. In the MSP1 and control groups, protection correlated with antibody to MSP3 raised by challenge infection. PMID- 11865424 TI - Age- and infection intensity-dependent cytokine and antibody production in human trichuriasis: the importance of IgE. AB - The cytokine and antibody response to Trichuris trichiura infection was determined for 96 persons living in an area where the parasite is highly endemic and infection exhibits a convex age intensity profile. In response to stimulation with T. trichiura antigen, a small proportion of the study group produced interleukin (IL)-4 (7%), IL-9 (5%), and IL-13 (17%). A larger proportion produced IL-10 (97%), tumor necrosis factor (TNF)-alpha (93%), and interferon (IFN)-gamma (32%). The levels of TNF-alpha (P =.016) and IFN-gamma (P =.012) significantly increased with age, suggesting a switch to a more chronic infection phenotype. The predominant parasite-specific antibodies produced were IgG1, IgG4, IgA, and IgE. Unlike the IgG subclasses and IgA, parasite-specific IgE correlated negatively with infection intensity, as defined by egg output (P =.008), and positively with host age (P =.010). These findings suggest a mixed cytokine response in trichuriasis and an IgE-associated level of protection. PMID- 11865425 TI - Leishmania-induced cellular recruitment during the early inflammatory response: modulation of proinflammatory mediators. AB - This study investigated whether Leishmania species, the etiologic agent of cutaneous (Leishmania major) and visceral (Leishmania donovani) leishmaniasis, could differentially elicit early inflammatory events in vivo correlating with the subsequent development of their reciprocal pathogenesis. By use of the murine air pouch system, injection of Leishmania led to a rapid and transient accumulation of a mixed population of leukocytes, and L. major recruited 31-fold more leukocytes than did controls, compared with 7-fold more leukocytes for L. donovani. L. major promastigotes were better than L. donovani promastigotes at inducing proinflammatory cytokine secretion and chemokine gene expression in pouch exudates. L. major infection elicited significantly increased chemokine receptor gene expression, compared with L. donovani infection. Collectively, the data reveal that L. major is a strong inducer of the early inflammatory response, compared with L. donovani, and suggest that such an immunologic event potentially could restrain this parasite to the inoculation site, favoring the development of local swelling and cutaneous lesions. PMID- 11865426 TI - Effect of aging on cytokine production in response to respiratory syncytial virus infection. AB - In vitro cytokine production in response to respiratory syncytial virus (RSV) and influenza infections was investigated in 11 "young" (mean age, 31 years) and "older" (mean age, 75 years) healthy volunteers by use of interferon (IFN)-gamma ELISPOT and ELISA analysis of cytokines in culture supernatants. Autologous dendritic cells (DCs), derived by culturing adherent peripheral blood mononuclear cells in granulocyte-macrophage colony--stimulating factor and interleukin-4, were used as antigen-presenting cells. Older subjects produced significantly fewer IFN-gamma ELISPOTs in response to RSV than the younger subjects. These results suggest that aging may be associated with a defect in the T cell response to RSV, even when DCs are used to maximize costimulation. This defect in cellular immunity may be related to the increased morbidity observed with RSV infection in elderly persons. PMID- 11865427 TI - Effect of previous or simultaneous immunization with canarypox expressing cytomegalovirus (CMV) glycoprotein B (gB) on response to subunit gB vaccine plus MF59 in healthy CMV-seronegative adults. AB - Development of a vaccine for prevention of congenital cytomegalovirus (CMV) disease is a priority. This study evaluated a "prime-boost" strategy by comparing the safety and immunogenicity of 3 doses of subunit CMV glycoprotein B (gB) vaccine plus MF59 (a squalene-in-water emulsion), 2 doses of a canarypox recombinant vaccine expressing CMVgB (ALVAC-CMVgB) followed by 2 doses of the subunit gB vaccine, 3 doses of both vaccines administered concomitantly, and placebo in 105 healthy, CMV-seronegative adults. Systemic adverse events were rare, but local reactions were common in all groups. After the first subunit vaccination, neutralizing antibody titers in the prime-boost group were comparable to those in subjects receiving 2 subunit vaccinations, indicating a priming effect of ALVAC-CMVgB. However, after the final dose, antibody and cell mediated immune responses were not significantly different among the groups. All 3 vaccine regimens induced high-titer antibody and lymphoproliferative responses, but no benefit for priming or simultaneous vaccination was detected. PMID- 11865428 TI - Genetic evidence of transmission of human T cell lymphotropic virus type 1 between spouses. AB - Sexual transmission of human T cell lymphotropic virus type 1 (HTLV-1) is considered to be an important route of infection in adults. However, no direct evidence has been reported that supports this observation. To address this issue, sequence variations of the gp46 (envelope)-coding region of HTLV-1 were determined in 13 patients infected with HTLV-1 who experienced seroconversion and in their spouses. Twenty-two nucleotide changes that were different from the reference sequence of lambdaATK-1 were identified. However, the gp46 sequences found were identical within each married couple. HTLV-1 proviral DNA loads measured in 11 of these couples varied from 10 to 3430 copies per 10(5) PBMC, and the proviral DNA loads of spouses often differed. This study provides the first genetic confirmation of the transmission of HTLV-1 from a carrier spouse to his/her partner. The findings also suggest that host-related factors play a more important role than do virus-specific factors in determining HTLV-1 proviral DNA load. PMID- 11865430 TI - Cocaine enhances human immunodeficiency virus replication in a model of severe combined immunodeficient mice implanted with human peripheral blood leukocytes. AB - Epidemiologic studies have identified cocaine as a cofactor for development of acquired immunodeficiency syndrome (AIDS). To evaluate this interaction, human peripheral blood leukocytes (PBL) were implanted into severe combined immunodeficient mice and infected with human immunodeficiency virus (HIV) in both the presence and absence of cocaine. Concurrent administration of cocaine resulted in significantly more PBL becoming infected with HIV in vivo (38.85% vs. 18.5%). The number of CD4(+) cells recovered from HIV-infected, cocaine-treated animals was significantly lower than that from mice infected with HIV in the absence of cocaine (6.5 x 10(4) vs. 19 x 10(4)) and was associated with a lower CD4:CD8 ratio and a dramatic increase in virus load. Exposure to cocaine alone did not affect the implantation of PBL, suggesting a specific interaction between cocaine and HIV. This report describes a model for evaluating HIV cofactors and supports cocaine's role in the development and progression of AIDS. PMID- 11865431 TI - Genetic basis of patients with bacille Calmette-Guerin osteomyelitis in Japan: identification of dominant partial interferon-gamma receptor 1 deficiency as a predominant type. AB - Interferon (IFN)-gamma-mediated immunity plays an important role in host defense against intracellular pathogens, especially mycobacteria. Six Japanese children with bacille Calmette-Guerin (BCG) osteomyelitis were evaluated (1 disseminated, 3 multiple, and 2 solitary types) for mutations of genes involved in interleukin 12-dependent, IFN-gamma-mediated immunity. Heterozygous small deletions with frameshift (818del4 and 811del4) that are consistent with the diagnosis of partial dominant IFN-gamma receptor 1 (IFN-gammaR1) deficiency were detected in 3 unrelated patients. Expression of IFN-gammaR1 on monocytes was significantly increased in all 3 patients. Screening of family members with recurrent and disseminated mycobacterial infections found the identical deletion in 1 of the fathers. Antimycobacterial treatment was effective in these patients and resulted in good clinical outcome. This study demonstrated that partial dominant IFN gammaR1 deficiency was the most common in Japanese patients who showed predisposition to curable BCG osteomyelitis. PMID- 11865429 TI - Prognostic value of the stromal cell-derived factor 1 3'A mutation in pediatric human immunodeficiency virus type 1 infection. AB - A mutation of the stromal cell-derived factor 1 gene (SDF-1 3'A) was shown to protect adults exposed to human immunodeficiency virus type 1 (HIV-1) from infection and to affect HIV disease progression in adults. The presence of this mutation in HIV-1-infected Kenyan children did not predict mother-to-child virus transmission. The SDF-1 3'A polymorphism was studied in 256 HIV-1-infected, 118 HIV-1-exposed but uninfected, and 170 unexposed and uninfected children of Italian origin, and the frequency of SDF-1 3'A heterozygosity and homozygosity in each of the 3 groups was similar. Of the 256 HIV-1-infected children, 194 were regularly followed up and were assigned to groups according to disease progression. The frequency of the SDF-1 3'A allele was substantially lower among children with long-term nonprogression than among children with rapid (P =.0329) or delayed (P =.0375) progression. We show that the presence of the SDF-1 3'A gene correlates with accelerated disease progression in HIV-1-infected children born to seropositive mothers but does not protect against mother-to-child HIV-1 transmission. PMID- 11865432 TI - A new viral agent, SEN virus (SENV), has been detected in patients from several countries: the pathogenic role of SENV in coinfections with hepatitis B virus or hepatitis C virus should be investigated. PMID- 11865433 TI - Mapping of a Plasmodium falciparum pfcrt K76T mutation: a useful strategy for controlling chloroquine resistance in Madagascar. PMID- 11865434 TI - The study of Mycobacterium leprae infection in interferon-gamma gene--disrupted mice as a model to explore the immunopathologic spectrum of leprosy. AB - Mycobacterium leprae infection was evaluated in interferon-gamma knockout (GKO) mice. At 4 months, growth of the bacilli in the footpads of GKO mice plateaued a log(10) higher than that in control mice. Control mice exhibited mild lymphocytic and histiocytic infiltrates, whereas GKO mice developed large, unorganized infiltrates of epithelioid macrophages and scattered CD4 and CD8 T cells. Flow cytometric analysis of popliteal lymph node cells demonstrated similar profiles of T cells; however, GKO cells exhibited an elevated proliferative response to M. leprae antigen. Expression of inducible nitric oxide synthase mRNA was decreased in GKO mice, whereas macrophage inflammatory protein-1alpha and interleukin-4 and -10 mRNA expression were augmented. Control and GKO activated macrophages inhibited bacterial metabolism and produced nitrite. Thus, although deficient in an important Th1 cytokine, GKO mice possess compensatory mechanisms to control M. leprae growth and feature elements resembling mid-borderline leprosy in humans. PMID- 11865436 TI - Listeria monocytogenes: clinical and experimental update. AB - Listeria monocytogenes, a small gram-positive bacillus, causes sepsis and meningitis in immunocompromised patients and a devastating maternal/fetal infection in pregnant women. Recent outbreaks demonstrated that L. monocytogenes can cause gastroenteritis in otherwise healthy individuals and more severe invasive disease in immunocompromised patients. Centralized processing in the food industry may be the cause of these large-scale listeriosis outbreaks. The mouse model of listeriosis, which was developed in the 1960s, has been extraordinarily useful for studying T cell-mediated immunity. Contrary to the original concept that macrophages are the principal effector cells in listeriosis, we found that immigrating neutrophils play the predominant role in early liver defenses. At later time points, CD8(+) T cells lyse infected hepatocytes by both perforin- and Fas-L/Fas--dependent mechanisms. Of interest, nonclassical major histocompatibility complex (MHC) class Ib--restricted cytolytic activity is expressed early during primary infection, whereas MHC class Ia--restricted activity is predominant through late primary and secondary infections. PMID- 11865435 TI - Infection and nitric oxide. AB - The author describes how his experience as an infectious disease fellow at Stanford University with Jack Remington from 1969 to 1971 influenced the direction of his subsequent laboratory investigation. The author reviews a series of studies that were intended to provide a mechanistic understanding of an in vitro cytotoxicity assay developed while he was a fellow with Jack Remington. These investigations resulted in the 1987 discovery of the synthesis of nitric oxide from L-arginine by cytokine-activated macrophages. This work provided the components (the precursor, products, and an inhibitor) of the enzymatic synthesis of nitric oxide by all three later-identified nitric oxide synthase isoforms. The significance of cytokine-induced nitric oxide synthesis during innate resistance and cell-mediated immune reactions is discussed briefly. PMID- 11865437 TI - The shikimate pathway and its branches in apicomplexan parasites. AB - The shikimate pathway is essential for production of a plethora of aromatic compounds in plants, bacteria, and fungi. Seven enzymes of the shikimate pathway catalyze sequential conversion of erythrose 4-phosphate and phosphoenol pyruvate to chorismate. Chorismate is then used as a substrate for other pathways that culminate in production of folates, ubiquinone, napthoquinones, and the aromatic amino acids tryptophan, phenylalanine, and tyrosine. The shikimate pathway is absent from animals and present in the apicomplexan parasites Toxoplasma gondii, Plasmodium falciparum, and Cryptosporidium parvum. Inhibition of the pathway by glyphosate is effective in controlling growth of these parasites. These findings emphasize the potential benefits of developing additional effective inhibitors of the shikimate pathway. Such inhibitors may function as broad-spectrum antimicrobial agents that are effective against bacterial and fungal pathogens and apicomplexan parasites. PMID- 11865439 TI - Approaches toward the directed design of a vaccine against Borrelia burgdorferi. AB - The overall efficacy of a recombinant vaccine for Lyme disease that is effective worldwide will depend upon the selection of one or more immunoprotective target(s) and the frequency of genetic variation, which can alter the antigenicity of the immunoprotective epitopes of the target proteins. Careful delineation of these protective epitopes on target antigens is essential for the development of vaccine candidates as well as for understanding the limitations of such vaccines. Structural models of these targets will provide critical information about conformation and specific residue surface accessibility for defining protective epitopes. Co-crystal structures with Fab fragments of protective antibodies will further delineate critical antigen surfaces. Population genetics will provide vital information on the heterogeneity of these proteins. Detailed epitope mapping will provide the information needed for the bioengineering of antigens needed to expand the specificity of a candidate vaccine. PMID- 11865438 TI - Epigenetic regulation of T cell fate and function. AB - During their development, T lymphocytes make sequential cell fate choices: T rather than B lymphocytes, then TCRalphabeta or TCRgammadelta, CD4 or CD8, and Th1 or Th2 lineage. These fate choices require the initiation of new programs of gene expression, and once initiated, these programs must be faithfully propagated in a heritable manner from parental cells to their progeny. With the exception of the T cell receptor, these changes in gene expression occur without a change in information encoded directly in the DNA sequence. Rather, these heritable programs of gene expression are imposed, at least in part, epigenetically through changes in chromatin structure and DNA methylation, allowing T cells to tune the threshold for expression of specific genes. PMID- 11865440 TI - Biotechnology and clinical trials. AB - Not surprisingly, clinical trials have been critically important to developments in the field of biotechnology. Perhaps less expectedly, the clinical trials of biotechnology products have been critically important to recent developments in the field of clinical trials design, conduct, and analysis. This manuscript explores three examples of biotechnology clinical trials--a trial in sepsis, a trial in fibrinolytics in myocardial infarction, and trials in gene therapy--and highlights their contributions to the theory and practice of clinical research. PMID- 11865441 TI - Host resistance in the brain against Toxoplasma gondii. AB - Interferon (IFN)-gamma-dependent, cell-mediated immunity plays the major role in resistance against development of toxoplasmic encephalitis (TE). Humoral immunity also participates in controlling Toxoplasma gondii in the brain. Resistance is operative under collaboration among T and B cells, IFN-gamma-producing non-T cells, microglia, astrocytes, and dendritic cells. A number of cytokines, including IFN-gamma, mediate interactions between these cells and activation of effector cells that prevent intracellular replication of the parasite. The L(d) gene confers resistance against development of TE in mice. In humans, the HLA-DQ1 and -DQ3 genes are involved in regulating the resistance and susceptibility. Since these genes are a part of the major histocompatibility complex, which regulates the immune responses, the regulation of the responses by these genes appears to be important for determining host resistance to this disease. Strains of T. gondii also affect development of TE. Genotypes of the parasite may be an important factor for determining development of TE. PMID- 11865442 TI - Suppression of NF-kappaB activation by infection with Toxoplasma gondii. AB - The interaction of host cells with microbial products or their invasion by pathogens frequently results in activation of the NF-kappaB family of transcription factors. The studies presented here reveal that in vivo, infection with Toxoplasma gondii results in the activation of NF-kappaB. To determine whether host cells could activate NF-kappaB in response to invasion by T. gondii, Western blots, immunofluorescence, and electrophoretic mobility shift assays were used to assess the response of host cells to infection. In these studies, infection of macrophages or fibroblasts with T. gondii did not result in the activation of NF-kappaB. In addition, the ability of lipopolysaccharide to activate NF-kappaB was impaired in cultures of macrophages infected with T. gondii. Together, these data demonstrate that invasion of cells by T. gondii does not lead to the activation of NF-kappaB and suggest that the parasite may actively interfere with the pathways that lead to NF-kappaB activation. PMID- 11865443 TI - Laboratory diagnosis of Toxoplasma gondii infection and toxoplasmosis. AB - For the past 40 years, the Toxoplasma Serology Laboratory at the Palo Alto Medical Foundation Research Institute (TSL-PAMFRI) has been dedicated to the laboratory diagnosis of Toxoplasma gondii infection and toxoplasmosis. TSL-PAMFRI is the "brain child" of Jack S. Remington. Jack's ceaseless devotion to objectivity and uncompromising excellence has made TSL-PAMFRI the Toxoplasma reference laboratory for the Centers for Disease Control and Prevention, the US Food and Drug Administration, and health care providers and clinical laboratories in the United States and other countries. Jack's leadership and vision created, defined, and significantly contributed to the development of laboratory methods for the diagnosis of the infection and diseases caused by T. gondii. A summary of the laboratory tests currently available at TSL-PAMFRI for the diagnosis of infection and disease caused by the parasite is presented here. PMID- 11865444 TI - CD154 and type-1 cytokine response: from hyper IgM syndrome to human immunodeficiency virus infection. AB - CD40 and CD154 (CD40 ligand) are surface molecules that are central to the cross talk between T cells and antigen-presenting cells. This article reviews the relevance of CD40-CD154 interaction for regulation of interleukin-12/interferon gamma production in response to Toxoplasma gondii as an example of an intracellular pathogen. The manner in which defects in CD154 signaling contribute to immunosuppression and susceptibility to opportunistic infections in patients with X-linked hyper IgM syndrome and in patients with human immunodeficiency virus infection is also discussed. PMID- 11865445 TI - Protective effects of immunization with a recombinant cyst antigen in mouse models of infection with Toxoplasma gondii tissue cysts. AB - A portion of the major Toxoplasma gondii tissue cyst antigen (MAG1) was expressed in bacteria as a fusion to glutathione S-transferase (GST) and the purified fusion protein (rMAG1) was used to immunize mice in an attempt to induce protective immunity against challenge with live cysts from the T. gondii ME49 strain. Sixty percent of mice immunized with rMAG1 and challenged with 500 cysts of the T. gondii ME49 strain survived, while only 20% of mice immunized with GST alone survived, suggesting a protective effect specific to the MAG1 portion of the recombinant protein. In a model of chronic infection with ME49 cysts, rMAG1 immunized mice had significantly fewer cerebral cysts and reduced inflammation in the brain compared with mice immunized with GST alone. PMID- 11865447 TI - Abstracts of the 8th European Meeting on Complement in Human Disease. 8-12 September 2001. Strasbourg, France. PMID- 11865446 TI - Oral infection of C57BL/6 mice with Toxoplasma gondii: a new model of inflammatory bowel disease? AB - Infection with Toxoplasma gondii is naturally acquired through the oral route by ingestion of undercooked or raw meat containing cysts of the parasite or through ingestion of contaminated water or food contaminated with cysts or oocysts. Following peroral infection with 100 cysts of the ME49 strain of T. gondii, C57BL/6 mice die within 13 days after infection, whereas BALB/c mice survive. At day 7 of infection, massive necrosis of the villi and mucosal cells in the ilea is observed in C57BL/6 but not BALB/c mice. CD4(+) T cells, interferon-gamma, tumor necrosis factor-alpha, and inducible nitric oxide synthase mediate the development of necrosis. These findings indicate a Th1-type immunopathology, with parasite replication appearing to be involved in the first 3 days of infection. Murine and human studies on the immunopathogenesis of inflammatory bowel disease (e.g., Crohn's disease) also indicate a Th1-type immunopathology. The shared and distinct features of oral infection of mice with T. gondii and murine models of inflammatory bowel disease are discussed herein. PMID- 11865448 TI - Priorities for research in complementary medicine. PMID- 11865449 TI - In silico mapping of mouse quantitative trait loci. PMID- 11865450 TI - Author and subject indexes. PMID- 11865451 TI - [Acute respiratory distress syndrome. Etiology, physiopathology, diagnosis]. PMID- 11865452 TI - [Non-cirrhotic alcoholic liver disease. Pathologic anatomy, diagnosis, development, treatment]. PMID- 11865453 TI - [Positive ionotropic substances (digitoxin and digoxin). Principles and rules of use]. PMID- 11865454 TI - [Tobacco advertising and public health]. PMID- 11865455 TI - [Discovery of the HLA system]. PMID- 11865457 TI - [Biology of the transmissible agent responsible for subacute spongiform encephalopathies]. AB - Transmissible spongiform encephalopathies, also called prion diseases, are brain degenerative ailments. These diseases, that are either infectious, or genetic, or sporadic in origin, are characterised by the accumulation of a host protein, the prion protein. Although the disease mainly implicates the brain, the immune system is involved in the propagation of the infectious agent which is always associated with the prion protein. This protein presents a normal structure in healthy brain but changes its conformation in association with the disease. It has been possible to reproduce some characteristics of this conversion in vitro. The protein only hypothesis is grounded on this conformational change which is correctly simulated by physico-chemical models. This hypothesis can explain most of the characteristics of prion diseases but the existence of an other element associated with infection cannot be ruled out. PMID- 11865456 TI - [Brief history of transmissible spongiform encephalopathies]. AB - Transmissible spongiform encephalopathies have a long and still unfinished history replete with spectacular discoveries and equally spectacular failures. The story begins in the 18th century with the first descriptions of scrapie, a naturally contagious disease of sheep that was not experimentally proven to be a transmissible infection until 1936. This discovery permitted veterinarian investigators to begin to explore aspects of pathogenesis and the nature of the infectious agent. The additional discovery, in the 1960's, that scrapie was similar to the human diseases kuru and Creutzfeldt-Jakob vastly widened what had been a comparatively restricted field of research, and led to rapid advances in molecular characterization of the disease agents, culminating in the proposal that they are self-replicating abnormally configured host proteins, or "prions". Despite these advances, we have seen in recent years 3 potentially preventable outbreaks of disease: iatrogenic Creutzfeldt-Jakob disease due to contaminated pituitary hormone extracts and dura mater grafts, and a new variant of the disease (Will-Ironside syndrome) linked to consumption of beef contaminated by the agent of bovine spongiform encephalopathy, underscoring the disparity between fundamental knowledge and its practical application. PMID- 11865458 TI - [Biopathology of transmissible subacute spongiform encephalopathies]. AB - Three lesions can be seen in the central nervous system: vacuolation ("spongiform state"), hypertrophy and proliferation of astrocytes ("astrogliosis"), and neuronal loss. These are poorly specific changes. In contrast, amyloid plaques and other deposits of PrPres are very specific lesions. The recent widening of the clinico-pathological spectrum of PrPres-induced disorders has shown that none of these lesions was constantly present. The mechanisms of PrPres occurrence and development, neuronal death, involvement of the central nervous system after peripheral inoculation are still hypothetical. B lymphocytes, monocyte macrophages, peripheral nerves, central synapses are important. New data will be provided by experimental models (transgenic mice, transplantations), and by a full study of all patients affected by prion diseases, which necessarily involves autopsy. PMID- 11865459 TI - [Creutzfeld-Jakob disease]. AB - Described in the early 1920s in Germany, Creutzfeldt-Jakob disease now comprises 4 entities: 1. familial forms are linked to mutations of the gene coding for prion protein (about 8% of the cases); 2. iatrogenic forms are due to interventions using contaminated material or human extracts (about 5% of the cases); 3. sporadic forms represent the majority of the cases (about 87%) without any established explanation; 4. recently described new variant, related to bovine spongiform encephalopathy, remains quite rare (41 known cases). In all its forms, the disease is constantly mortal after a usually short course. Major signs are rapidly evolving; dementia with myoclonus, ataxia, and electroencephalographic abnormalities (periodic activity). Formal diagnosis is histologic. PMID- 11865460 TI - [Genetics of human subacute spongiform encephalopathies]. AB - Variations in PRNP, the gene that encodes PrP in humans, play a major role in the genetic predisposition to prion diseases. Mutations in PRNP are found in patients with familial forms and are considered as causative. A coding polymorphism in PRNP (129 Met/Val) defines a predisposing factor to sporadic and acquired forms. Other favouring genes have been searched without real success. Both animal models and cellular models have been constructed to study the molecular mechanisms implied in the disease genesis. PMID- 11865461 TI - [Transmissible subacute spongiform encephalopathies in animals]. AB - Transmissible subacute spongiform encephalopathies are a group of neurodegenerative diseases of man and of several species of domestic mammals. Scrapie has been recognised in sheep and goats for more than 250 years. Descriptions in 1986 of the first cases of bovine spongiform encephalopathy (BSE) in United Kingdom, its expansion with an enzootic pattern, and in 1996 of a new variant of Creutzfeldt-Jakob disease, probably related to the same agent, focused the attention of scientists, medias and public on what seems to be a new zoonosis. The biopathological study of BSE and scrapie is useful for evaluating what risk these animal diseases represent for human health. PMID- 11865462 TI - [Risk prevention of transmissible subacute spongiform encephalopathies]. AB - The risk assessment of transmissible spongiform encephalopathies (TSEs or prion diseases) can be managed at three levels: 1. control of source material that could be potentially infected (biological products of human or animal origin, instruments in contact with these products); 2. control of the processes for the purification of biological products and instrument decontamination techniques; 3. evaluation of invasive medical or surgical acts. Highest infectivity is found in central nervous system. TSE agents do resist to almost all usual decontamination procedures. National recommended procedures include sodium hypochlorite 18,500 ppm or sodium hydroxide 1N for 1 hour and autoclaving at at least 134 degrees C for at least 18 min. PMID- 11865463 TI - [Therapeutic prospects for subacute transmissible spongiform encephalopathies]. AB - There is currently no effective therapy available for Creutzfeldt-Jakob disease. However, a limited number of drugs such as polyanions, the amyloid-binding dye Congo red, amphotericin B and anthracyclines have been found to delay the appearance of the clinical signs in experimental prion diseases. Today, the most promising agent appears to be less toxic derivative the amphotericin B, MS-8209. Indeed this compound has a wide spectrum of anti-prion activity and constitute the unique molecule able to prolong survival time when treatment is performed at the late stages of infection. This result represents an important progress in therapeutical approaches of prion diseases and justify the development of new polyene antibiotic derivatives. PMID- 11865464 TI - [Contempo 1998: the current status of telemedicine]. PMID- 11865465 TI - [Algodystrophy. Etiology, diagnosis, development, treatment]. PMID- 11865467 TI - [Thrombopenia. Diagnostic approach]. PMID- 11865466 TI - [Child services. Diagnosis, medical-legal guidance]. PMID- 11865468 TI - Handmaidens no more. PMID- 11865469 TI - Crisis in the classroom. AB - For once, the experts agree: there's trouble in the classroom. As Globe and Mail writer Jeffrey Simpson noted in a February 1999 column, experts have projected Canada will need at least 10,000 nursing graduates per year by 2011, compared to around 5,000 now coming out of RN diploma and basic baccalaureate programs each year. "Unless nursing schools beef up enrolments and nursing graduates can find employment, the mean age for nurses will keep on rising," he wrote. "...Finding, training and financing those thousands of additional nurses will be among the health-care system's most urgent demands in the years ahead." PMID- 11865470 TI - Recruiting nurses for the new millennium. AB - The challenge of finding the very best candidates to become the nurses of the future is not new. Even in the mid-1800s, Florence Nightingale struggled to seek out the best prospects for training at her new institutions. Today, however, with nursing colleges and universities in competition for the top candidates to fill their entrance quotas, recruitment has become not just a challenge but an art. PMID- 11865471 TI - Focus on function in geriatric assessment and rehab. AB - In 1994, the province of Alberta, driven by the need for deficit reduction, began a major restructuring of the provincial health care system. An integral component of this new health structure was accountability for service delivery, including the use of outcome measures to determine service effectiveness. This paper describes the efforts of one program to evaluate the effectiveness of its service delivery, in terms of functional patient outcomes, in the midst of restructuring and budget constraints. PMID- 11865473 TI - Nurse defends work of legal network. PMID- 11865472 TI - [The challenge of interdisciplinarity]. AB - The restructuring of health services and the shift to ambulatory care has resulted in several institutions considering the adoption of program management. This approach intensifies interdisciplinary team work, the latest care delivery method for various services. But what exactly is an interdisciplinary team? And what is the nurse's role within that team? These two questions are often asked in the literature, as well as in practice. The answers to these questions are still confusing. The inability of the nursing profession to define itself clearly in comparison with other health care professionals has negative consequences. This article presents a vision of interdisciplinarity as a means for nurses to define themselves within an interdisciplinary team. PMID- 11865474 TI - Questioning what we call "alternative". PMID- 11865475 TI - Examining life's value. PMID- 11865476 TI - The sexual assault nurse examiner. PMID- 11865477 TI - Nehinaw (Cree) understandings of diabetes. "He travels in the body". PMID- 11865478 TI - Therapeutic conversations that count. PMID- 11865479 TI - Girl child vulnerability. PMID- 11865480 TI - Preparing for expanded roles. PMID- 11865482 TI - Questionable ethics. PMID- 11865483 TI - Meaning found. PMID- 11865481 TI - Exposure to body fluids. PMID- 11865484 TI - Coping with neurofibromatosis. PMID- 11865485 TI - Refining your research question. PMID- 11865486 TI - The Manitoba Pediatric Cardiac Surgery inquest report. PMID- 11865487 TI - Regional planning study. Assessing the population with leg and foot ulcers. PMID- 11865488 TI - To stay or to go? Students' perspective on the recruitment and retention of nurses. PMID- 11865489 TI - Growing concern. Indoor air quality in Canadian schools. PMID- 11865490 TI - Perioperative nurses connect with families. PMID- 11865491 TI - Screening and tracking latex allergies. PMID- 11865492 TI - The home care dilemma. PMID- 11865493 TI - Moist healing versus wet-to-dry. PMID- 11865494 TI - Mental illness. Cloaked in secrecy and shame. PMID- 11865495 TI - Wellness. Healthy employees are good for business. PMID- 11865496 TI - Psychoeducational program. Adjusting to life with an ostomy. PMID- 11865497 TI - Urban core. "Home" nursing of HIV-positive clients. PMID- 11865498 TI - When CPR is not an option. AB - What is to be learned from this situation? First, although Mr. Lockwood's consent for the DNR order is not needed, there is an obligation to communicate openly and clearly with the family and ensure that Mrs. Lockwood's advance directive is respected. This might mean a DNR order needs to be written. Also, there is an obligation to discuss goals of care with the family. The second lesson is that you should reflect on your employer's CPR policies and practice, and ask the following questions: Do the policy and/or practices support saying "no" in a situation such as Mrs. Lockwood's? Also, how does the policy support staff when there is a request for futile CPR, either from a competent patient or from a patient's family? What are the expectations about communication with the family when there is an advance directive and/or when CPR is found to be futile? Knowing what you ought to do for patients is not sufficient. Often you cannot act on these decisions because of the environment. If the policies are not in accord with the CNA Statement on Resuscitative Interventions, you should collaborate with colleagues to revise the CPR policy and practices. By doing so, you will be meeting your obligation to help foster and support a practice environment that promotes ethical, competent and compassionate nursing care. PMID- 11865500 TI - Vitamin E and pruritus. PMID- 11865501 TI - End-of-life care in old age. PMID- 11865502 TI - Work concerns, family concerns. PMID- 11865503 TI - Nursing skills enhancement. PMID- 11865504 TI - Growing old with a developmental disability. PMID- 11865505 TI - High-risk antepartum patients. No place like home. PMID- 11865506 TI - Sleep and shiftwork. I. PMID- 11865507 TI - Nurses' experience of violence. PMID- 11865508 TI - Applauding lowry. PMID- 11865509 TI - Facing a worthy struggle. PMID- 11865511 TI - Why wear gloves? PMID- 11865510 TI - On breastfeeding. PMID- 11865512 TI - Researcher needed: please apply. PMID- 11865513 TI - Advocacy 101. Nursing leadership: unleashing the power. PMID- 11865514 TI - A regional approach to continence management. PMID- 11865515 TI - Universal newborn hearing screening programs. PMID- 11865516 TI - Complementary and alternative medicine: critical issues of nursing practice and policy. PMID- 11865517 TI - Pinpointing the causes of nurses' stress. PMID- 11865518 TI - Advancing breastfeeding. PMID- 11865519 TI - Older adults and advance care directives. PMID- 11865520 TI - Immigrant women's health. A different reality. PMID- 11865521 TI - Managing postoperative pain at home. PMID- 11865522 TI - An incredible learning journey. PMID- 11865523 TI - When parents and children disagree about care. PMID- 11865524 TI - Abuse unacceptable. PMID- 11865525 TI - A journey in professional growth. PMID- 11865526 TI - [Nephrolithiasis prophylaxis]. PMID- 11865527 TI - [HCV-HIV co-infection: a new clinical challenge]. PMID- 11865528 TI - [Blessed Josemaria Escriva's view on pain. Meeting on pain, open to the entire Medical School of Rome: faculty, students, administrators, etc...]. PMID- 11865529 TI - Effect of water of Anticolana Valley on urinary sediment of renal stone formers. AB - An investigation was carried out to ascertain the effect of drinking Fiuggi water on the microcrystalline structure of the calcium oxalate monohydrate present in urinary sediments provided from patients suffering from recurrent idiopathic oxalic calculosis. The experimental group was administered tap and Fiuggi water for ten days. The control group was administered tap and Fiuggi water according to the same procedure as for the experimental group. The comparative data show that drinking Fiuggi water leads to a strong reduction, and sometimes even to the elimination, of the calcium oxalate monohydrate present in the urinary sediment reducing the risk of oxalic calculosis. Fiuggi water contains organic molecules belonging to the fulvic acid family. These acids are capable of complexing the calcium ions and interact preferentially with the crystal lattice of the calcium oxalate monohydrate via the formation of a film and behave as pumping systems by linking the calcium ion, demolishing the crystal lattice and dissolving calcium and oxalate ions. Mineral water treatments must therefore be viewed as a function of the specific composition of the water administered. The ecosystem influences the composition of water, as a complex matrix containing a number of organic molecules which are potentially biologically active. PMID- 11865531 TI - Heart rate stationarity in heart transplanted patients. AB - The heart rate stationary was studied by tachographic of 24 Holter analysis in 15 normal subjects and in 13 recipient subjects that were heart transplanted at least 5 years and that, at the time of our research, presented a very good post operative course. To test the stationary heart rate we performed a scanning of Holter in qualified strips of four beats in which the three consecutive intervals demonstrate a constant acceleration or deceleration sequential variations of tachographic values. The results obtained demonstrated that in normal and transplanted subjects stationary and non stationary strips are evident. Both in normal and in transplanted subjects non-stationary strips are prevalent, although in different ways: in normal subjects the stationary and non-stationary ratio is 1:1.40; in transplanted subjects the ratio is 1:1.68. Non-stationary strips, that in the cartesian plane do not demonstrate any directional variation, in normal subjects are, on average, higher than in transplanted subjects. The same phenomenon is available for the strips with only one variation. The strips with three consecutive variations are much more evident (more 50%) in transplanted subjects. These strips are also more numerous compared to the sum of all the other strips with a single variation. The statistical analysis demonstrates that the difference between the normal and the transplanted subjects is significative. Our data can suggest an absent autonomic nervous system regulation and can confirm the results we obtained in these patients using a phase space analysis of the same Holter recording. PMID- 11865532 TI - [The medical mystery of hepatitis C]. PMID- 11865530 TI - [Daily sensation of hunger, before and after phototherapy, in subjects with depression-type seasonal affective disorder]. AB - OBJECTIVE: The present study investigates the daily pattern of hunger sensation (HS) in women affected by "seasonal affective disorder--depression type" (SAD DT), before and after therapeutic exposure to bright light (phototherapy). The aim is to detect whether there are disorders in daily HS during the active phase of the disease which can be normalized by an effective treatment of the depressive status, via phototherapy. MATERIALS AND METHODS: Volunteered for the study 4 women affected by SAD-DT, 32-58 years old (BMI: 17.8-29.6 Kg/m2); 10 clinically healthy women (CHW), 21-52 years old (BMI: 23 e 25 Kg/m2) were recruited as controls. Both of the SAD-DT patients and CHW were asked to compile the "orexigram", which was chronobiometrically analyzed by means of the 1. conventional statistical methods; 2. rhythmometric analysis; 3. spectral analysis. RESULTS: Before phototherapy, the SAD-DT patients were found to be characterized by an increased HS (hyperorexia), with the circadian rhythm of the orectic stimulus (OS) which is shifted in its acrophase, being prone to become "free running". After phototherapy, the SAD-DT patients were found to show a little insignificant decrease in their OS, which still maintains a delayed phase in its circadian rhythm as well as the tendency to be "free running". CONCLUSIONS: The pre-treatment findings suggest that the SAD-DT patients are affected by hyperorexia associated with a "phase-shift" for the circadian periodicity of their HS, which is prone to the desynchronization. Such a dyschronism reinforces the hypothesis that the SAD-DT may be pathogenetically sustained by a mechanism of "phase-shift". The post-treatment findings suggest that both the hyperorexia and dyschronism of the orectic circadian rhythm are uncorrected by the phototherapy, even though the SAD-DT patients seem to have had beneficial antidepressive effects from the therapeutic intervention. The persisting dyschronism indicates that the photic stimulus is not able to completely reset the biological clock of the suprachiasmatic nuclei, at least for the phasic modulation of the HS circadian rhythm. The orexigram, thus, could be enclosed among the clinical tools in order to assess the complete efficacy of the phototherapy in SAD-DP patients. PMID- 11865533 TI - [Pott disease: current diagnostic and therapeutic approaches]. AB - Spinal tuberculosis, or Pott's disease, has been known since ancient times; it was a common orthopaedic illness until 1950, but the progress of specific therapies confined it to third word countries; its resurgency in recent years in developed countries is due to drug resistance, immigration from endemic areas, and an increase in immunodeficient patients. Spinal disease is found in about 1% of all tuberculosis cases. Modern treatment consists of a diagnostic phase, based on clinical findings and imaging exams, and of a medical and surgical therapy. Our patients have been treated with a three antibiotic drugs scheme, and underwent early surgical treatment, in order to assure the curettage of the lesion and obtain spine stabilization, so as to avoid spinal deformity. PMID- 11865534 TI - [Role of magnetic resonance in shoulder disease]. AB - Next to the knee, the shoulder is the most common joint to be referred for MRI. Excellent soft tissue contrast and multiplanar acquisition provide optimal assessment of muscle, tendons, hyaline and fibrous cartilage, joint capsule, fat, bursae and bone marrow. In this article the most common indications for shoulder MRI are reviewed and discussed, but we focused primarily on the rotator cuff syndrome and shoulder instability. Correct diagnosis requires the use of appropriate pulse sequences and imaging planes, proper patient positioning, and a satisfactory surface coil. Moreover, technical improvements continuously augment the ability of MRI to study the shoulder; for example Magnetic Resonance arthrography is superior to the other imaging techniques in evaluation of glenohumeral joint. This interdependence between technical development in MRI and clinical advance in shoulder therapy ensures that MRI will continue to play an important role in the routine management of patients with shoulder disease. PMID- 11865535 TI - [The flu syndrome: update and therapeutic and prophylactic approach]. AB - Authors in the present work analyse the history of the flu from V century A.C. until our days. We can know the different flu epidemic the structure of the virus and to take note of the different preventing device; of these the core is the vaccine. Thanks the vaccine is possible to weaken the big epidemics. Authors attract the importance on the mass vaccine and conclude to mention the new antivirus drugs per os. PMID- 11865536 TI - [Central serous chorioretinopathy: a new extragastric manifestation of Helicobacter pylori?: Analysis of a clinical case]. AB - Central serous chorioretinopathy is classically characterized by a shallow, round, serous detachment of the macular retina. It preferentially affects men between 20 and 45 years of age and recurrences have been documented in most cases. Although the aetiopathogenesis of the disease is still unknown, it has been thought to be due to a focal leakage from one or more defects in the retinal pigment epithelium, which allow serous fluid from the choriocapillaris to diffuse into the subretinal space. Moreover, it is theorized that damage of the active fluid transport mechanisms through the retinal pigment epithelium may also play a contributing role. Finally, the reported correlation with psychophysical stress provides support for the concept of the disease being "adrenergically conditioned". Recently, an interesting association has been observed between this chorioretinal pathology and the Helicobacter pylori infection and, consequently, a new etiological hypothesis has been proposed. Here we report a retrospective analysis of a clinical case of central serous chorioretinopathy with Helicobacter pylori positivity. PMID- 11865537 TI - [Molecular treatment of cancer. Imatinib]. PMID- 11865538 TI - [What place for BCG vaccination in France?]. PMID- 11865539 TI - [Should infant walkers be forbidden?]. PMID- 11865540 TI - [From painful pronation of the elbow to severe hemophilia: an unusual step]. PMID- 11865541 TI - [Epidemiology and management of malnutrition in hospitalized children 0-5 years of age in Dakar]. PMID- 11865542 TI - [Integrated treatment of iron deficiency, vitamin A deficiency and intestinal parasitic diseases: impact on Senegalese children's growth]. PMID- 11865543 TI - [Clinical study and prevalence of fetal alcohol syndrome in medico-social institutions of the Reunion Island]. AB - BACKGROUND: Fetal alcohol syndrome (FAS) is a major problem in the Reunion Island and the Public Health Authorities decided to determine its prevalence in their medico-social centers on 31 December 1996. MATERIAL AND METHODS: A questionnaire was established to identify affected patients in the 20 medico-social centers in charge of 1320 children. Eighty-eight children were selected and 87 could be analyzed. RESULTS: Sixty-four of 87 (76.3%) were FAS and 23 of 87 (23.7%) had closely alcohol-related diseases. The prevalence was between 7.1 and 14.1% and lower than expected from available data. CONCLUSION: The study allowed to precise the social and familial factors predisposing to alcohol addiction during pregnancy. A TV prevention message will be broadcasted after this study. PMID- 11865544 TI - [Power Doppler sonography and acute pyelonephritis in children: comparison with Tc-DMSA scintigraphy]. AB - Acute pyelonephritis is a common infection in children. The clinical and biological diagnosis is still sometimes difficult. For most authors, Technecium 99m dimercaptosuccinic acid scintigraphy is considered as the gold standard tool for diagnosis but it is invasive and expensive. The aim of our study was to compare the sensitivity and the specificity of B-mode sonography and power doppler to DMSA-Tc scintigraphy in acute pyelonephritis. PATIENTS AND METHODS: Forty-nine children were enrolled in this study with suspicion of pyelonephritis. All infants underwent doppler sonography and scintigraphy within 48 hours after their hospitalization. Doppler sonography criteria were increased kidney size, thickness of sinus wall, vascular defect, and various echogenicity of the kidneys (focal or diffuse hyperechogenicity or focal hypoechogenicity). RESULTS: Among 28 children with a positive scintigraphy, 15 had a positive doppler sonography (sensitivity 54%) and 13 had a negative doppler sonography. Among 21 children with a negative scintigraphy, 20 had a negative doppler sonography (specificity 95%) and one had a positive doppler sonography. CONCLUSION: In clinically suspected acute pyelonephritis, doppler sonography has a high specificity. A positive doppler sonography should avoid the use of scintigraphy. PMID- 11865545 TI - [Prevalence of asthma in children in Auvergne: a safe region?]. AB - The aim of our study was to determine the prevalence of asthma in the Auvergne region. PATIENTS AND METHODS: Two thousand and nine pupils from primary (five-six years old) and secondary (14-15 years old) schools were given a questionnaire when attending their compulsory medical examination and asked to return it completed. RESULTS: The response rate was 82%. The prevalence of asthma was 5.6%. There was no difference neither between the two age groups nor between sexes. In contrast, there was a significant difference in prevalence according to whether the child lived in a town (7.3%) or in the countryside (4.6%) (p = 0.01). In the department of the Puy-de-Dome, this difference was even more distinct, with a prevalence of 10% in urban areas and 4.0% in the countryside (p = 0.001). Twelve percent of the children had experienced wheezing over the previous 12 months. The prevalence of bronchitis was higher in rural areas (17.3%) than in towns (11.8%) but not to a significant degree. Passive smoking significantly increased this risk (18.9% and 14%, p = 0.003). CONCLUSION: The prevalence of asthma in the Auvergne region is low in both age groups compared to other regions in France and to Europe. The difference of prevalence between town and countryside may evidence of the influence of environmental factors. PMID- 11865546 TI - [Progressive multifocal leukoencephalopathy revealing AIDS in a 13-year-old girl]. AB - CASE REPORT: A 13-year-old girl with an immunosuppression developed a right hemiparesis and a cerebellar syndrome. She was seropositive for HIV. A progressive multifocal leukoencephalopathy (PML) was suspected because of white matter lesions (MRI) and established on detection of JC virus in CSF by polymerase chain reaction (PCR). In spite of highly active antiretroviral treatment (HAART) and cidofovir, she died three months later. CONCLUSION: Late AIDS diagnosis was established during a neurological complication, 13 years after a neonatal transmission. PML caused by JC opportunistic virus still has a poor prognosis. PMID- 11865547 TI - [Kingella kingae: a rare cause of meningitis]. AB - Kingella kingae is the second most frequent germ involved in arthritis affecting young children. This germ isolation on ordinary environment is difficult, which may explain why it is seldom known. It is now widely accepted that a direct inoculate of articular and osseous samples on liquid substrate improves the culture sensitivity. Other septic localizations have been described such as endocarditis or, less commonly, meningitis. CASE REPORT: We report the observation of a five-year-old child, treated for meningitis, with CSF culture showing evidence of scarce colonies of Kingella kingae. CONCLUSION: By analogy with arthritis, Kingella kingae may regularly be undetected, not being isolated, in some cases of non-documented meningitis with a cerebrospinal fluid (CSF) cytology recalling a bacterial origin. It would be of interest to verify if the seeding of CSF in liquid substrate would increase the sensitiveness of the cultures. PMID- 11865549 TI - [Sudden death of a two-year-old boy with influenza A virus infection: study of an autopsy case]. AB - Influenza A virus infections are common in childhood and infancy and are often underdiagnosed while serious or lethal forms are rare. CASE-REPORT: We describe a case of sudden death in a two-year-old boy. Pathologic findings at autopsy were consistent with Myxovirus influenzae A virus infection and the virus was isolated by post mortem PCR. CONCLUSION: In the case of sudden death in infants, especially if pathologic findings are compatible with a viral infection, PCR may allow identification of the causative virus. PMID- 11865548 TI - [Therapeutic approaches in children immunized with vaccine Prevenar]. PMID- 11865550 TI - [Group B streptococcal infection with 2 recurrences in a newborn]. AB - Group B III streptococcus (GBS) is a predominant pathogen in neonates in France. Relapse is rare and two successive relapses are exceptional: only three cases have been yet reported. CASE REPORT: A newborn infant of 18 days of age presented a first episode of invasive group B III streptococcal infection with meningitis and ventriculitis. At 53 days of age, a second episode with bacteriemia and parotidis appeared. At 63 days of age, she presented a third episode with meningitis. Genome analysis of the three bacterial strains isolated during the three episodes showed the same clonal origin. COMMENTS: We discuss the incidence of the treatment, the eventual presence of a penicillin-tolerant GBS, the possible relapse or recurrence of the pathogen and the role of the relative immunodeficiency in infant. PMID- 11865551 TI - [Neurofibromatosis 1: recommendations for management]. AB - Twenty experts, members of a French medical network devoted to neurofibromatosis 1 have elaborated recommendations for the management of the disease. Bibliography was obtained through a Medline of articles from 1966 to 1999 for the terms neurofibromatosis, NF1, neurofibroma and from textbooks. A consensual document was written taking into account extracted data. An annual careful clinical examination is recommended except in cases with complications. Screening investigations are not recommended due to the rarity of complications, generally symptomatic and easily detected during the clinical follow-up. The only controversial exception might be magnetic resonance imaging for early detection of optic pathway gliomas in young children. A co-ordinated follow-up in specialised multidisciplinary centres, providing patients with a rational management, is recommended. PMID- 11865553 TI - [Treatment of subclinical congenital toxoplasmosis by sulfadiazine and pyrimethamine continuously during 1 year: apropos of 46 cases]. AB - In France, most of children suffering from congenital toxoplasmosis have an infraclinic or moderate type at birth. This study aimed at evaluating, on the mid term, tolerance and results of postnatal treatment previously given in severe toxoplasmosis. METHODS: A retrospective study considered 46 children with a mild or moderate congenital toxoplasmosis treated over 12 months with sulfadiazine pyrimethamine and treatment was completed since three months. RESULTS: Five children suffered from a lesion of chorioretinitis during treatment and two after. After a mean follow-up of 27.1 months, ten children (21.7% 95%CI [12.1 35.9]) had at least one ocular injury. Specific IgG titers and immune load were diminished to become almost non-existent at the end of the year of treatment (respectively p < 10(-5) and p = 0.0005). No thrombocytopenia was observed. Twenty-three children (50%) had at least one episode of neutropenia < 1000/mm3, 14 had only one, nine presented two or more installment. None was followed by an infection. CONCLUSION: This therapeutic pathway is more demanding but shorter than those usually offered when associating pyrimethamine-sulfadiazine. Yet, it does give identical result on the mid term. Longer follow-up is needed to appreciate. Active molecule on cysts should be introduced. PMID- 11865552 TI - [Nosocomial infections due to human coronaviruses in the newborn]. AB - Human coronaviruses, with two known serogroups named 229-E and OC-43, are enveloped positive-stranded RNA viruses. The large RNA is surrounded by a nucleoprotein (protein N). The envelop contains 2 or 3 glycoproteins: spike protein (or protein S), matrix protein (or protein M) and a hemagglutinin (or protein HE). Their pathogen role remains unclear because their isolation is difficult. Reliable and rapid methods as immunofluorescence with monoclonal antibodies and reverse transcription-polymerase chain reaction allow new researches on epidemiology. Human coronaviruses can survive for as long as 6 days in suspension and 3 hours after drying on surfaces, suggesting that they could be a source of hospital-acquired infections. Two prospective studies conducted in a neonatal and paediatric intensive care unit demonstrated a significant association of coronavirus-positive nasopharyngal samples with respiratory illness in hospitalised preterm neonates. Positive samples from staff suggested either a patient-to-staff or a staff-to-patient transmission. No cross-infection were observed from community-acquired respiratory-syncitial virus or influenza infected children to neonates. Universal precautions with hand washing and surface desinfection could be proposed to prevent coronavirus transmission. PMID- 11865554 TI - [Recent progress in pediatric neurological rehabilitation]. AB - New challenges in rehabilitation for children with neurological diseases directly depend on advances made in medical research and on the quality of the environment. This is relevant to motor function as a whole, to new therapeutic avenues in spasticity, to global approaches in the evaluation of cognitive and learning disabilities, as well as curative perspectives in neuromuscular disease. Networking with the family and other actors in the environmental field is essential to achieve a better social integration. A true collaboration between physicians and pediatricians is necessary to work toward more progress. PMID- 11865555 TI - [Mental retardation and environment: contribution of growth curves to etiological diagnosis]. AB - Although deprived environment is known for more than 50 years to be one of the etiology of mental retardation in infancy, this remains unrecognized by many paediatricians and family practitioners. Yet if appropriate therapeutic measures are instituted early enough, future development of the child is good. A study of the growth curves in relation with the environmental changes, can help to diagnose environmental mental retardation. As an illustration, we present the cases of 5 children first addressed for developmental delay, and secondly removed from their usual environment for psychosocial reasons. Mean duration of follow-up was 10 years, including at least 5 years after the removal. All 5 children had non-organic failure to thrive which was displayed only on the retrospective study of their records. Four recovered from their developmental retardation. PMID- 11865556 TI - [Contraception and voluntary termination of pregnancy: legislative measures derogatory to parental consent in France]. AB - Several derogatory clauses to parents' consent have been recently introduced in the French legislation concerning adolescents contraception and voluntary termination of pregnancy. Thus, from now on, adolescents may have free and anonymous access to contraception as well as detection and treatment of sexually transmitted diseases. Adolescents may also obtain emergency contraception without medical prescription. Parents' consent is no more necessary to authorize voluntary termination of pregnancy for an adolescent who wish to keep it secret. PMID- 11865557 TI - [Treatment of febrile convulsions]. AB - Information and reassurance of parents are essential before considering therapeutic measures following a first febrile seizure. It must be emphasized that, although impressive, febrile convulsions are essentially benign, and that in the majority of cases there will be no recurrence. Apart from that, therapeutic measures are of limited efficacy in order to prevent reocurrence. The correct use of antipyretic treatment during febrile episodes must be specified. Due to its several inconveniences, oral diazepam as an intermittent prophylaxis should be only considered in case of multiple reoccurrences. There is little place for continuous anticonvulsant treatment. PMID- 11865559 TI - [Post-traumatic Korsakoff's syndrome: clinical and anatomical report]. AB - Clinical and anatomical report of a post traumatic amnestic syndrome (Korsakoff's syndrome), associating anterograde amnesia persisting for fifteen years, with temporal disorientation, false recognitions, initially intense transitory confabulation and secondarily bursts of confabulation, intact remote memory and persistence of old learnings. Death after twelve years from mesenteric infarction. Anatomically, post-traumatic sequellae are limited to both cingulate gyri in their anterior part. This cingula involvement is easy to understand if one knows that post-traumatic Korsakoff's syndrome appears after severe cranial traumatisms, with at least three days of coma, and develops constantly, even if transitory, after long duration comas with 20 to 30 days of loss of consciousness. Anatomical explanation depends on the fact that Korsakoff's syndromes from various etiologies need, to be produced, a bilateral damage of the limbic circuit and that severe head traumatisms, when dying early in the evolution without possibility of a neuro-psychological investigation, have always a destruction of corpus callosum or cingulate gyri or both, resulting from crushing of these structures by the edge of the faulx cerebri. Consistent with these constatations, it is logical that a Korsakoff's syndrome develops after severe head traumas with bilateral lesions of the limbic circuit and especially of the cingulate gyri. But anatomical evidence remains rare, because early fatal evolution does not permit psychological evaluation and, reversely, long survivals who may die from another pathology would not have brain examination. We prefer the name of "Korsakoff's syndrome" rather than that of "amnesic syndrome" to denominate the anterograd amnesia (amnesie des faits recents) encountered in nutritional disorders due to B1 deficiency in true Korsakoff's disease, but also with other etiologies such as cerebral tumours, vascular cerebral disorders, post commital anoxia, herpetic encephalopathy, head traumas, all of them developing amnesia for recent events, formerly classified under the title of "korsakowian syndrome" or "mental syndrome of Korsakoff" and more recently under the denomination of "amnesic syndrome". But whatever is the etiology of the memory disorder, the amnesic syndrome remains identical and the advanced small differences, such as euphoria in alcoholics or mood depression in tumours, are often fallacious, so that the only way of differentiation deals with accessory symptoms such as intracranial hypertension in tumours, sudden onset in vascular etiologies or polyneuritis in B1 deficiency. Post-traumatic Korsakoff's syndrome joins with this scheme, for its clinical aspect is so similar to that of nutritional disorders that it might be difficult to reach the exact diagnosis when an alcohol addict develops, after a head trauma, an amnesia which could be the consequence of the trauma but also of a nutritional disorder developed after the accident with inadequate parenteral treatment. Our case, which is the first well documented observation of this disease reported with long clinical survey and final pathological examination, was presented in 1981 at a joint meeting of the French and Dutch neurological societies. It gives the proof of the importance, in limbic circuit, of the cingulate gyri. A comparison is made with four other clinical cases of post traumatic amnestic syndrome with MNR procedures which show, for two of them, cingula lesions explaining the clinical features, for one of them a bilateral lesion of Ammon's homs and for the last one extra limbic lesions, with destruction of the inferior part of both frontal lobes, associated with a possible deafferentation of the right cingula cortex. PMID- 11865558 TI - [Psychiatric and psychological aspects of premenstrual syndrome]. AB - Numerous, but heterogeneous studies have been performed about premenstrual syndrome, with finally a lack of credibility and interest among practitioners. More recently with the diagnosis criteria generalization, psychiatrists were more concerned about this syndrome, because of anxiety and mood symptoms involved in social impairment and need of medical care. In 1983 in the United States, the National Institute of Mental Health conference devoted to this topic proposed the first diagnosis criteria, requiring a prospective and daily assessment of the symptoms. In 1987, the American Psychiatric Association, in the DSM III-R, introduced the Late Luteal Phase Dysphoric Disorder diagnosis that became in 1994 in the DSM IV the Premenstrual Dysphoric Disorder, with the same diagnosis criteria. In the literature, prevalence rates are very heterogeneous according to the diagnosis criteria used and to the populations studied. One of the most relevant criteria is the induced impairment, such as avoidance of social activities, or search for medical care. Lifetime prevalence is thus estimated between 75 and 85% if considering the report of one or several symptoms, between 10 and 15% in case of medical care request, and between 2 and 5% in case of social activities interruption. To distinguish isolated complaints from a disabling disorder, self-questionnaires are the best way of assessment in a so complex and changing disease. Most of the epidemiological studies found a positive correlation between the premenstrual dysphoric symptoms and the lifetime major depressive disorder diagnosis. However, recent prospective studies failed to find an association between premenstrual syndrome and an increased risk of major depression. On the other hand, some studies showed that the premenstrual period is a risk period for associated psychiatric disorders exacerbations, as the obsessive-compulsive disorder, more severe alcohol intakes in case of alcoholism, symptoms increase in schizophrenics, or higher rates of suicide attempts. The most widely studied and frequently blamed etiopathogenic hypothesis is the serotonin dysregulation. Serotonin is particularly involved in expression of irritability and anger, but also in occurrence of depressive symptoms and specific food cravings, precisely found in the premenstrual dysphoric disorder. Among their different effects, estrogens increase the density of serotonin receptors and enhance the sensitivity to serotonin agonists. Moreover, some studies found a significantly different response to d-fenfluramine, a serotonin agonist, in women with premenstrual dysphoric disorder. In psychoanalytical theories the premenstrual syndrome was associated to a "femininity complex", to an ambivalent pregnancy desire, and to unconscious conflicts relating to sexual preference. In this context, Karen Horney, who took a great interest in the premenstrual period, was radically opposed to the Freudian theory of feminine sexuality, in particular the negation of the female sex. For Karen Homey, the "desire of penis" is more expressive of the woman's spite not to share the sexual, but also political, social and cultural benefits fallen to men. To understand the premenstrual period feelings it is also necessary to take into account the personal history of the woman and the psychosocial factors involved, as the social and cultural beliefs, and the mother-daughter communication. Medical cares are necessary when symptoms constitute a severe and disabling disorder. Among non-psychiatric treatments, progesterone was the most widely prescribed treatment, but relating to recent performed studies, it failed to prove its efficiency in such an indication. In the same way, the efficiency of the contraceptive pill was not demonstrated. The most prescribed psychiatric treatments are serotonin re-uptake inhibitors and benzodiazepines. First studies showing serotonin re-uptake inhibitors efficiency in premenstrual dysphoric disorder were performed in the beginning of the nineties, with clomipramine and fluoxetine, and later fluvoxamine, paroxetine, sertraline and citalopram. Studies having compared the efficiency of antidepressants according to their serotonin activity (paroxetine or sertraline versus maprotiline, that is a selective noradrenaline re-uptake inhibitor), showed that serotonin re-uptake inhibitors were significantly more efficient on all symptoms than maprotiline, that was not more efficient than placebo. Low doses of clomipramine (10 to 50 mg per day) seem to be sufficient and it appears also preferable to prescribe an intermittent treatment because of a possible tolerance effect, susceptible to be warned by phases free of treatment. Alprazolam was the most studied benzodiazepine in this indication. Most studies were positive, using daily posologies of 0.25 to 4 mg during the 6 days preceding the menses, with improvement of irritability, anxiety and depressive mood. The general practitioner frequently carries out psychological support, in particular in case of mild symptoms without consequences. Nevertheless, underestimate a more severe psychological suffering is a risk, firstly because there is no systematic interrelationship between the somatic symptoms intensity and the psychological distress, and secondly because premenstrual period is a special emotionally moment to put in evidence psychological or relational disruption. All kinds of psychotherapy can be relevant, even though the training of relaxation techniques is particularly suitable in such an indication. In conclusion, and in spite of the generalization of the diagnosis criteria in the international psychiatric classifications as the DSM, the premenstrual syndrome remains a complex and polymorphous disorder. The premenstrual syndrome was considered for a long time like a somatic disease, but now the psychiatric symptoms severity justifies most often the medical cares. In order to distinguish some isolated and mild complaints, of a disabling disorder, the standardized prospective auto-assessment is the most relevant method. Finally, intermittent prescription of serotonin re-uptake inhibitors appears to be the most effective treatment, the previously used hormonal treatments not having made proof of their efficiency in such an indication. PMID- 11865560 TI - [French version of TASTE (test for the ability and evaluation)]. AB - Ability to study and evaluation is only one example of performance among many others but research and publications concerning this issue for more than 50 years, especially in the context of test anxiety and need of achievement, conferred upon it a prototypical dimension. Investigations about motivation also stimulate many scientists and constitute another foundation of this study (13). The level of performance depends on knowledge and motivation (33). Time devoted to study is essential to succeed; so motivation and procrastination are in competition. The importance of reinforcement (extrinsical motivation) and the desire for learning and knowing (intrinsical motivation) are determinant. Other elements must be emphasized: guarantee of obtaining rewards, self efficacy and causal attribution. These considerations point out the multidimensional and interactive aspects of test anxiety (7, 31). The number of components is not described unanimously but experts agree with emotional, cognitive and behavioral dimensions (25). So, anxiety was approached in its motivational properties, and it was the case until the sixties, in terms of drive corresponding to a need like thirst or hunger (18); then it was conceptualized in a dynamic context broader than that of stress and coping (29, 30). Last, it constitutes the object of theories highlighting cognitive interference (9, 23, 26) or defective skills (8, 32). A lot of questionnaires were built without answering the different aspects and for instance without linking the theoretical and therapeutic components concerning this problem. Committed to the traditional fields of research (test anxiety and need of achievement), to Weiner's work about attribution theory (34) and that of Bandura in self efficacy (4, 5), E. Depreeuw (10) was particularly interested in Heckhausen's model (16, 17), trying to associate experimental conceptions with the clinical reality. On this basis, he elaborated the TASTE (10, 12, 20): test for ability to study and evaluation. At first constituted of 121 items, the self-questionnaire, after factorial analysis, was reduced to 78 items assessing 4 factors which represent the 8 components of Heckhausen's model (16). The first factor (30 items) concerns anxiety in its emotional and cognitive characteristics. Interesting data were observed by Depreeuw and confirmed in the Netherlands and in Greece (19), especially the fact that anxiety of girls is higher than that of boys when they are confronted with an exam. The second one (19 items) represents self-confidence: confidence in ability to succeed and on using adequate strategies. The third factor (14 items) squared with value attributed to performance. It especially comments on the intrinsical component of motivation. The fourth factor (15 items) corresponds to procrastination; study is postponed on behalf of activities which actually reduce the achievement process. The self-report questionnaire (where answers run from total disagreement to total agreement whether they correspond to the way of thinking or acting) takes account of the emotional, cognitive and behavioral dimensions of the model. The multidimensional aspect of this questionnaire, but mainly its connections with theory and clinic, are convincing; we adopted it with other tests in research about fear of failure. Validation of the French translation is the subject of this article. In order to realize this operation, we chose the initial version of TASTE (121 items) to assess a Belgian sample of french-speaking students in the first year of University of Liege (n = 617). They are differentiated by gender, faculty (Economy, Medicine, Philosophy, Psychology, Applied Sciences) and experience of failure, i.e. the fact of repeating a school year. Statistics were realized with SPSS and SAS. A comprehension test and a back translation are satisfactory. The stability over time too: the one week test-retest was achieved with a sample of 33 student nurses; comparing the factors two by two, intra-class correlations ranged from 0.5 to 0.95. The component analysis with Varimax rotation does not allow us to find the four factors of construction. We obtain the same disappointing results with the version of 78 items. According to the screentest, we adopted the solution of five factors which confirms the original construction. A fifth particularly strong factor (Cronbach's alpha 0.82) corresponds to devalorisation. The internal reliability is very satisfactory. If we consider the items strong loading (more than 0.30), the French version is constituted of 94 items. If we consider the items which are specifically loading each factor, we can reduce the questionnaire to 50 items. Internal reliability remains the same. Correlations between the data obtained with this brief version are satisfactory; comparing the factors in pairs. Pearson's coefficients range from 0.8 to 0.89. The study of E. Depreeuw was realized ten years ago with a Belgian sample of Flemish-speaking students; is the cultural context or an evolution in test anxiety which explains such a difference in the results? We are now assessing a sample in another country which used the TASTE in 1996 in order to have some information about comparison of samples. In any case, the questionnaire of E. Depreeuw respects the multidimensional aspect of test anxiety. Our choice of 5 factors according to principal components confirms the original structure of the test but also discriminates another factor which could be predictive of psychopathology (panic, depression). This new dimension measures negative perception of ability and achievement especially when compared with other students. One aim of such a study is to discern clusters of students i.e. groups of students with characteristics such as Covington (7), Depreeuw (20) and Zeidner (35) describe them. This description is very important; the French version is only a part of a very broader study including other tests and could differentiate student's profiles. The French version of 5 factors finally gives results in agreement with the original. The items (10 per factor) are selected according to their highest specificity and after elimination of redundancy. Validity remains in the short version, making it more useful in clinical practice. PMID- 11865562 TI - [Eye tracking disorders in schizophrenic patients and their parents]. AB - Several studies have confirmed the existence of genetic factors in schizophrenia. However, the genotype predisposing for the disease is not known yet. Nevertheless, those genetic factors in the families of schizophrenic patients urge us to search for genetic vulnerability markers of schizophrenia. Ocular pursuit disorders, in particular, could be one of those vulnerability markers. Eye movements have been often tested in schizophrenia. Most of the schizophrenic patients have eye-tracking disorders and their biological relatives demonstrate an increased prevalence of eye-tracking impairments. The aim of the study was to research if smooth pursuit eye movements could be a vulnerability marker of schizophrenia. In order to have an indication about this hypothesis, impairments of smooth pursuit eye movements were researched in both schizophrenics and their parents. METHODS: Fifteen DSM IV schizophrenic patients stabilized at the time of the inclusion and not treated with lithium, benzodiazepines, barbiturates, or chloral hydrate; 19 parents without history of schizophrenic spectrum disorders (SADSLA and IPDE), and 2 groups of healthy subjects matched in age and sex with probands and with the parents, were included in the study. Parents only were included (fathers or mothers) in order to have an homogeneous population for the genetic risk and age. The eye-tracking paradigm used was a smooth pursuit task. The stimulus was a sinusoidal wave form moving on a horizontal line, with a frequency of 0.4 Hz and an amplitude of 30 degrees. Different parameters were measured: gain (ratio between the eye velocity and the target velocity) and saccades frequencies (catch-up saccades, back-up saccades, anticipatory saccades and square-wave-jerks). For each parameter, analysis of covariance (ANCOVA) with age as covariable was carried out. For the results reaching the significance of 0.05, the Bonferroni correction was applied (level of significance 0.016). The effect size of the parameter was calculated ((the mean of the subjects minus the mean of the matched controls) divided by standard deviation of the two groups). According to Cohen, 0.20 indicates a small effect size, 0.50 indicates a medium effect size and 0.80 indicates a large effect size. RESULTS: Comparison between patients and matched controls: the means of global gain, of gain for the movements to the left and of gain for the movements to the right did not differ significantly between patients and their matched controls. The size effects are 0.31 for the global gain, 0.20 for the movements to the left and 0.41 for the movements to the right. The frequencies of total saccades, catch-up saccades, back-up saccades, anticipatory saccades and square-wave-jerks did not differ significantly between patients and their controls. The size effects for those parameters were 0.09, 0.03, 0.00, 0.39 and 0.63 respectively. Comparison between parents and matched controls: the means of global gain, of gain for the movements to the left and of gain for the movements to the right did not differ significantly between the two groups. The size effects for those parameters were 0.00, 0.05 and 0.17 respectively. The frequency of total saccades did not differ significantly between the groups whereas the size effect was 0.63. The frequency of catch-up saccades was significantly more important in parents than in controls (p = 0.006) and the size effect was 0.80. The other saccadic parameters did not differ significantly between groups, their size effects were 0.24 for the back-up saccades, 0.21 for the anticipatory saccades and 0.00 for the square-wave-jerks. Whereas the gain of the patients had a tendency to be lower than the gain of their controls, no significant difference was observed between patients and their controls. Only a size effect of 0.63 for the frequency of square-wave-jerks was obtained. This large effect size suggests that the difference between patients and controls might be significant in a larger sample. The catch-up saccades frequency between parents and controls was significant. The differences between our study and the previous studies could be due to several factors. The paradigms used were different between the studies and our sample was small (only 15 patients and 19 relatives). Moreover, some patients in the previous studies were treated by lithium, drug well known to modify ocular pursuit and, finally the relatives in the other studies were 10 years older than ours and age is known to alter ocular pursuit. Since an impairment of the smooth pursuit was observed in the relatives of schizophrenic patients but not in the probands, this study does not support the hypothesis that eye-tracking disorders could be considered as a marker of vulnerability of schizophrenia. PMID- 11865561 TI - [Current and lifetime prevalence of obsessive compulsive disorders in eating disorders]. AB - A significant proportion of patients suffering from Eating Disorders (ED) present a comorbidity with anxiety disorders. Among the anxiety disorders, Obsessive Compulsive Disorders (OCD) are the third most frequently diagnosis observed in ED. However, prevalence rates from the literature are contradictory depending on the diagnostic criteria and evaluation tools used. Studies concerning the chronology of appearance of OCD and ED and the role played by denutrition are even rarer and equally contradictory. OBJECTIVE: The aim of this study is to bring new empirical data to the study of the correlations between OCD and ED by exploring, in a significant clinical sample, the current and lifetime OCD comorbidity in the diagnostic sub-groups and sub-types of ED as defined by the DSM IV and to study the chronology of appearance of these disorders taking into account the role played by denutrition. We make the assumption that there should be a difference in the prevalence of obsessive compulsive disorders in the various ED sub-groups and sub-types and that purging anorexics should, at equivalent levels of denutrition, exhibit higher OCD prevalence rates than the other sub-types as a result of their more severe general psychopathology. METHODS: Current and lifetime prevalence were investigated using the Mini International Neuropsychiatric Interview (MINI) and the Yale-Brown Obsessive Compulsive Scale in a sample of 89 DSM IV eating disorders in and out-patients aged between 15 and 30 (58 AN and 31 BN) and 89 matched controls. In order to increase the validity of the current diagnosis of OCD only patients presenting an OCD diagnosis on the MINI (excluding obsessions and compulsions related to food and body image) and a score of 16 or more on the Y-BOCS were included in the study. RESULTS: Current and lifetime prevalence of OCD in ED is significantly higher than in general population (15.7% and 19% vs 0% and 1.1%, p < 0.05). Anorexic and bulimic patients do not show any difference in the current and lifetime comorbidity (19% and 22.4% vs 9.7% and 12.9%, ns). Concerning the diagnostic sub-types, following our assumption purging anorexics present the higher current and lifetime prevalences (29% and 43%), followed by restrictive anorexia (16%) and purging bulimia (13%). The only significant difference is found in the lifetime prevalence of OCD between purging and non purging anorexics. In the great majority of cases (65%) OCD diagnosis precedes ED diagnosis and OCD current prevalence and Y-BOCS scores of underweight patients are not significantly higher than normal weight patients, suggesting that there are only limited links between denutrition and obsessionality. LIMITS: Some methodological limitations must be considered. First of all, the small sample size of the diagnostic sub-types of ED do not enable us to draw definitive conclusions concerning the prevalence of OCD in the clinical forms of ED. Some of the diagnostic sub-types of ED, as BN-NP, appear at different ages: in order to better investigate the prevalence of OCD in all the diagnostic sub-types, larger age ranges should be considered. Secondly, our sample of anorexics is almost made of hospitalized inpatients recrutated in a specialized university unit, whereas bulimics are all consulting outpatients. It is possible that the higher than normal rates of OCD in anorexics could be related to the severity of this sample. Comparisons with samples of consulting anorexics should be performed. Lastly, it is necessary to evoke the limitation represented by the choice of a healthy control group. OCD are rare in the general population and the differences found between the clinical and the control groups do not offer strong arguments to support the specificity of the association between OCD and ED, which would be better explored by using a pathological control group. CONCLUSIONS: So far, none of the various models proposed to explain the comorbidity observed between ED and OCD appears completely satisfactory. ED and OCD would share a genetic vulnerability to a dysregulation of serotoninergic functioning which would predispose these subjects, depending on specific clinical and biological conditions, to develop an obsessional and compulsive symptomatology more or less focused on food and body image. The knowledge of the clinical and biological specificities of the ED diagnostic sub-types presenting an OCD comorbidity could point the way towards specific therapeutic interventions in these patients. Our study of the comorbidity of OCD in ED makes a further contribution to the identification of specific links between the OCD and the various clinical forms of ED. More clinical and biological studies are needed to further explore these relationships. PMID- 11865563 TI - [Study of dissociative disorders and depersonalization in a sample of young adult French population]. AB - Questioned by several researches about dissociative disorders, the authors study differences established on the nosographic register, through a quantitative study and a psychodynamic argumentation in a sample of french population. From the utilisation of the Dissociative Experiences Scale (DES) created by Bernstein E and Putnam FW (1986), which is an excellent screening tool for dissociative disorders and constructed on DSM II diagnostic criterions, the authors will show the interest of a psychodynamic analysis of dissociative disorders, in the face of the diagnostic difficulty in relation to several approaches of this concept. This difficulty is studied giving the background to dissociative disorders and depersonalization. Ionescu (1999) shows that between 1890 and 1910 dissociation represents one of major themes of psychology, psychopathology and psychiatry. Then, this interest about dissociation decreases and will be almost non-existent in the middle of the twentieth century. The interest for dissociative disorder will grow in the eighties with north-american studies about multiple personality disorders. Until 1980, dissociative disorders exist in DSM II as a list of symptoms included into hysterical neurosis, among the conversive disorders. In 1980, the publication of DSM III replaces the notion of hysteria with the notion of dissociative disorder. In this way, we can see on the one hand somatoform disorders quarterly corresponding to the ancient version of conversive hysteria, and on the other hand dissociative disorders characterized by a perturbation of consciousness, memory, identity or perception of environment. In 1994, The DSM IV delete the notion of hysteria and neurosis and keeps only the notion of dissociative disorders. They include now the five following categories: dissociative amnesia, dissociative fugue, depersonalization disorder, dissociative identity disorder, dissociative disorder not otherwise specified (including derealization). Depersonalization disorders consist of "persistent or recurrent episodes of depersonalization characterized by a feeling of detachment or estrangement from one's self. The individual may feel like an automation or like he or she is living in a dream or movie" (DSM IV). Depersonalization disorder cannot be diagnosed if it is part of schizophrenia, panic disorder, acute stress disorder or dissociative identity disorder. Various depressive disorders, hypocondriasis or obsessive-compulsive disorders can accompany depersonalization disorder. The first purpose of this study will search the frequency of dissociative disorders and depersonalization in a sample of normal population. Further, the inclusion of depersonalization amongst dissociative disorders seems not so evident: depersonalization belongs to self-consciousness disorder in french psychiatry. This fact seems more logical insofar as dissociative disorders have all together a memory and consciousness perturbation, and this perturbation is missing from depersonalization's feeling. The second purpose will be to clarify and specify the particularity of depersonalization among dissociative diorders, from the psychopathological point of view. METHODOLOGY: The sample (n = 248) is made up of french young adults aged 17 to 30 (mean age = 20, SD = 15 and 24% is male population). Subjects were streamming from universities. The screening tool which was used is the Dissociative Experiences Scale, a 28-item patient questionnaire regarding various dissociative symptoms. The subject is asked to indicate the percentage of time, to the nearest 5%, that particular symptom is experienced. The score is made by adding the various percentages and finding a mean that is expressed in numbers from 0 to 100. Normal scores are in the range of 5 to 15 in american adults. RESULTS: The utilization of principal component analysis (PCA) with varimax rotation is justified by the will to compare this study with American's studies. The mean score obtained is 17.44%, and 13.3% of the scores exceed a psychiatric threshold at 30%. The descriptive analysis shows that the component 1 (PCA without varimax rotation) represents 33.02% of total explained variance. This result demonstrates that the structure of the DES is based on one concept, the same as the american population, it is the concept of dissociation. The Principal Component Analysis with varimax rotation of the DES ratings yielded a tree-factor solution: imaginative absorption (F1), depersonalization-derealization (F2) and dissociative amnesia (F3). Mean score for each factor is respectively: F1 = 21.56%, F2 = 13.95%, F3 = 11.04%. DES reliability was studied through computation of Cronbach's coefficient (0.92). The PCA with varimax rotation brings to the fore a full dissociative disorder without any trouble of memory and consciousness. This fact questions again once more the link between hysteria and dissociative disorders. There is here a clinical distinction between depersonalization-derealization and other dissociative disorders. Indeed, the absence of significant alteration of memory and conscience is specific of depersonalization and derealization in this study. CONCLUSION: Finally, this study concurs with DSM IV dissociative criterions. At last, one factor of PCA is composed by the association of depersonalization and derealization, in contradiction with DSM IV definition. This result shows that, into the french population, we cannot divide the two concepts. PMID- 11865564 TI - [Restraint and seclusion in psychiatry: review and prospects]. AB - In this article, the authors reviewed the literature published since 1965 concerning restraint and seclusion. They synthesized the contents of the articles reviewed using the categories of indications and contraindications; rates of seclusion and restraint as well as demographic, clinical, and environmental factors that affect these rates; effects on patients and staff; implementation; and training. The literature on restraint and seclusion supports the following: 1) Seclusion and restraint are basically efficacious in preventing injury and reducing agitation; 2) It is nearly impossible to operate a program for severely symptomatic individuals without some form of seclusion or physical or mechanical restraint; 3) Demographic and clinical factors have limited influence on rates of restraint and seclusion; 4) Training in prediction and prevention of violence, in self-defense, and in implementation of restraint and/or seclusion is valuable in reducing rates and untoward effects; 5) Studies comparing well-defined training programs have potential usefulness. PMID- 11865565 TI - [Interest of propranolol in the treatment of school refusal anxiety: about three clinical observations]. AB - School refusal anxiety is a pathopsychological disorder which touches the young child, between 8 and 13 years. Even if the school refusal is studied for a long time, there is not still consensus as for the specific definition of this disorder or on the best way of treating it. Nevertheless, accountable of long lasting difficulties in school integration, its short and medium term consequences are serious and well known: school desertion, mood disorder and behavioral problems. Speed and quality of the medico-psychological and educational interventions represent a important factor for evolution and prognosis. Although, psychological interventions remain essential, sometimes the interest of an associated psychotropic medication should be discussed. This one can indeed either improve their results or supporting their installations. Despite more than twenty controlled trials in the pediatric population, no definitive psychopharmacological treatment data exist for anxiety disorder in childhood and especially for school refusal disorder. The majority of the studies stress as well the interest of benzodiazepines as tricyclic antidepressants but without being able to specify the possible superiority of a chemical on the other. On the other hand, the side effects of each one are well-documented, in particular for the benzodiazepines (potential abuse, sedation, potential desinhibition, mnemonic disorder), limiting thus their uses in child. In this work, we would like to emphasize the interest of propranolol in the treatment of somatic symptoms usually met in school refusal anxiety. Although beta-blockers have been used in the treatment of neurovegetative symptoms associated with situational anxiety disorders, there is no controlled data and only some open data to guide pediatric use for anxiety disorders in children. Nevertheless, prescribed with low posology and in substitution of benzodiazepine, this medication enabled us in three severe clinical cases to shorter notably the time of school rehabilitation. Well tolerated on the clinical level, with a greater efficiency on the somatic signs related to anxiety than benzodiazepines and with not having their side effects, this therapeutic can constitute a significant support in the psychological treatment of these children. However, these present results require to be confirm by other observations, which will be lead perhaps to a controlled study. PMID- 11865566 TI - [Confusion and delusions occurring during interferon-alpha treatment]. AB - A number of side effects occurring during IFN therapy have been described. Psychiatric side effects include cognitive, behavioural and affective changes, confusion and, less commonly, psychotic symptoms. We report the case of a 52-year old patient who had been treated for almost three years with IFN alpha for a chronic myelogenous leukemia and who presented an acute confusional episode with delusions, following the reintroduction of IFN alpha after a therapy suspension of 2 weeks. Clinical improvement occurred quite rapidly after cessation of the IFN treatment; this evolution and the normality of the biological tests, EEG and cerebral imagery suggest a causal relationship between this treatment and the psychiatric symptoms. PMID- 11865568 TI - Demography is destiny. PMID- 11865567 TI - [Psychiatric manifestations of lupus erythematosus systemic and Sjogren's syndrome]. AB - We present one case of Sjogren's syndrome (SS) secondary to systemic lupus erythematosus (SLE) with predominant psychiatric manifestations, treated with success by cyclophosphamide. From this case, we review the psychiatric aspects of these two autoimmune diseases as described in the literature and we present the etiopathogenic hypothesis and treatment of the psychiatric disorders. Case report -In August 1996, a 38 year old man was admitted in our psychiatric department for agitation. Primary SS had been diagnosed in July 1996. He had previously attempted to suicide but was never hospitalized in a psychiatric department. During the hospitalization in our department, the patient had auditive hallucinations and felt persecuted. He received loxapine 400 mg/day and was remitted in a few days. He was discharged to a convalescent home with the diagnosis of brief psychotic disorder. In October 1996, he was readmitted to our department for agitation. He had shown agitated behavior and aggression in the convalescent home. There were no hallucinations and no affective disorders. He became calm rapidly and was discharged home a few days later. In November 1996, he was found in a coma by a neighbor. He was admitted to an intensive care unit. The lumbar punction revealed blood cells. Cerebral computer tomography showed subarachnoid hemorrhage. The diagnosis was meningeal hemorrhage due to vasculitis. After regaining consciousness, the patient complained of reduced visual acuity. This was believed to be due to retrobulbar neuritis and the patient's vision improved slightly with corticosteroids. The third hospitalization in our department occurred in February 1997 for depression. The patient had shut himself away for days in his apartment. He had suicidal ideas. His mood improved progressively under fluoxetine 40 mg/day. He was discharged to a convalescent home with the diagnosis of major depressive disorder. The fourth and last admission in our department occurred in June 1997. There were disturbances of memory and orientation. He felt sad and guilty about accusation of sexual abuse on his daughter. He presented typical histrionic symptoms: he had catatonic attitudes only in public areas such as the corridors. Cerebral computer tomography and electroencephalogram were normal. There was no biological abnormality. Signs of confusion rapidly disappeared. He felt better after reintroduction of fluoxetine 40 mg/day. Diagnosis was non-specified depressive disorder, but this episode could be retrospectively seen as delirium. After being hospitalized on these four occasions in one year in our psychiatric department, the diagnosis of his systemic disease was revised by rheumatologists. The patient was diagnosed as suffering from systemic lupus erythematosus associated with secondary Sjogren's syndrome. From September 1997, he received cyclophosphamide 2 g intraveinously per month during 6 months. His vision improved dramatically. His ocular dryness became milder. His mood is now stable. He has not suffered from hallucinations or delusion since. Psychiatric disorders in SLE--During the course of SLE, the occurrence of psychiatric manifestations varies widely from 5 to 83%. They include psychotic disorders, major depressive disorders, subtle cognitive disorders and personality disorders of histrionic type. Etiopathogenic hypothesis are: direct activity of the disease on the central nervous system by autoantibodies (antiphospholipide and antiribosome P autoantibodies) (18, 19) or cytokines (interleukin 2, interleukin 6, alpha interferon) (38, 59), side-effects of glucocorticosteroids and hydroxychloroquine (16) or anxious reaction to a chronic and potentially lethal illness (43, 54). Nevertheless, immunologic and cerebral imagery research suggests that psychiatric disorders are related to vasculitis and non-inflammatory vasculopathy of the small cerebral blood vessels. The management of the patients should include treatment of the disease itself and specific psychotropic treatment. Glucocorticosteroids and especially intravenous infusions of immunosuppressive agents, such as cyclophosphamide, are effective. Psychotropic drugs must be used, making sure to avoid SLE-inducing drugs, like chlorpromazine, carbamazepine and lithium carbonate (19, 20, 45). In addition, psychologic care is essential. Psychiatric disorders in SS--During the course of the primary SS, the occurrence of psychiatric disorders is large as well: from 20 to 70% (47, 61, 62). They are mainly major depressive disorders, anxiety disorders, cognitive disorders and dementia. Brief psychotic disorders and delirium are rare. Etiopathogenic hypotheses are similar as those in SLE, with some differences: antiphospholipide and antiribosome P autoantibodies are not usually found in SS and anti-Ro (SSA) autoantibodies in serum are associated with psychiatric disorders (3-11, 61). According to Drosos et al. (29, 30), psychiatric disorders are explained by psychological distress. This slowly progressive fluctuating disease creates constant discomfort from dysphagia, dyspareunia and functional disability. Some of these manifestations can be treated by corticosteroids and psychotropic drugs. Drugs with anticholinergic side-effects, like phenothiazines, tricyclic antidepressants and hydroxyzine which can enhance the oral dryness have to be avoided. Social and psychological support is important too. DISCUSSION: The diversity of psychiatric morbidity in SLE and SS may be due to differences in patient selection and a lack of uniform clinical criteria. Studies which use standardized diagnostic criteria and control groups don't allow one to come to a conclusion about the relative prevalence of the psychiatric disorders in these autoimmune diseases. This will probably be resolved thanks to the recently published "American College of Rheumatology nomenclature and case definitions for neuropsychiatric lupus syndromes" (1). Finally, we can ask ourselves if there is a significant number of undiagnosed SLE and SS in psychiatric departments. Two studies report systematic search for SLE in psychiatric patients. In 1992, Hopkinson et al. (39) searched for several autoantibodies in serum samples of nearly 300 hospitalized psychiatric patients. In 1993, Van Dam et al. (65) did the same with more than 2,000 patients admitted to a psychiatric hospital. Hopkinson et al. found 1% undiagnosed SLE, which is much higher than in general population, and recommended to search SLE in every patient with a high erythrocyte sedimentation rate in psychiatric services. Results of the Van Dam et al. study suggest on the contrary, that SLE is not a common cause of admission to psychiatric hospitals. There is no study which report systematic search of Sjogren's syndrome in a psychiatric department. This is probably because most of patients receive or have recently received psychotropics with anticholinergic side-effects which is an exclusion criteria of SS. CONCLUSION: Psychiatrists should keep in mind that SLE and primary SS are potential causes of psychiatric manifestations when examining patients with multiple unexplained somatic complaints and psychiatric symptoms. They should then search for autoantibodies in the serum after careful physical examination. Diagnosis of SLE or SS could lead to a better adapted prescription of corticosteroids and/or immunosuppressive drugs and specific psychotropic drugs, making sure to avoid lupus-inducing drugs in SLE and drugs with anticholinergic effects in SS. The existence of psychiatric manifestations in SLE and SS constitutes an indisputable clinical reality that each practitioner must be able to recognize and treat. PMID- 11865569 TI - Deaf college students' reading comprehension and strategy use. AB - Two comprehension studies were conducted with 46 deaf college students. In the first, 20 deaf college students representing higher and lower reading-ability levels were tested for correctly stating the main idea of a passage, answering content questions, indicating their understanding of the words and phrases, and recognizing a topically incongruent sentence embedded in the passage. The results suggest that deaf students profess a better understanding of what they read than they are able to demonstrate. The students' inability to identify a topically incongruent sentence in the passage further suggests a need for them to more carefully and accurately evaluate their understanding of what they are reading. A second study investigated the effect of strategy review instruction on deaf college students' comprehension of short reading passages. Students reading at a higher level showed improved comprehension on the posttraining passage, but students reading at a lower level did not. Similarly, the control group of deaf students comparable to the higher-level readers did not show improved comprehension. PMID- 11865570 TI - A rationale and recommendations for sexuality education in schools for students who are deaf. AB - The authors evaluate and advocate the need for comprehensive sexuality education that meets the unique needs of youth who are deaf or hard of hearing, while calling for the expansion of teacher preparation in this critical area. Effective comprehensive sexuality education is designed to prepare young people to become more comfortable with, and informed about, their sexuality. Teachers and parents are key adults in this process. However, the responsibility for preparing teachers to handle sexuality education lies with both the postsecondary teacher preparation program and the administrative team at the individual school; their willingness to provide comprehensive training, current resources, and continued support are crucial to the success of any comprehensive sexuality program. In the individual school, effective guidance of youth who are deaf or hard of hearing in making appropriate decisions about their sexuality is built upon a team that includes not only school staff, but also parents and deaf adults in the community. PMID- 11865571 TI - A "consumer" survey of educational provision for deaf and hard of hearing students. AB - The researchers report the results of a survey of 140 deaf and hard of hearing students attending integrated or self-contained high school classrooms in the state of New South Wales, Australia. The survey was designed to gather information about educational experiences and preferences for various types of educational placements. There were no gender differences in placement, but there were hearing status differences (81.6% of the students in segregated placements had severe or profound hearing losses, compared to 52.4% of the students in integrated settings). Even though students were aware of the advantages and disadvantages of the different placement options, 80% indicated that they were generally satisfied with their current placement. One implication is that a range of options seems to be more appropriate than a one-size-fits-all model, at least from students' perspectives. The views and preferences of students should be considered when educational provisions are designed for students who are deaf or hard of hearing. PMID- 11865572 TI - The limits of narratives in understanding teacher thinking. AB - Researchers have argued that narrative provides insight into teachers' thinking and a model for the storage of knowledge about teaching. Concurrently, however, there are numerous cautions about using narratives as data sources. The present study addresses two problems: the limits of narrative as a data source and the feasibility of productively analyzing narratives. It also addresses the question of whether teachers actually store information as narratives. For the study, 23 deaf or hearing teachers of the Deaf participating in a project on integrating technology into teaching were interviewed about their experiences as teachers in general and in using technology in the classroom in particular. They rarely generated stories spontaneously. Rather, responses were related to the nature of the stimulus question. For example, when asked about their "worst class," teachers did not provide complete narratives but instead gave responses containing problems without resolutions. The study results suggest that teachers do not store information about teaching as narratives, but nonetheless can expertly construct narratives when given the right opportunity. PMID- 11865573 TI - Interpreting in mental health settings: issues and concerns. AB - Sign language interpreters in mental health settings face extreme linguistic and cultural difficulties in interpreting basic, everyday language used in these settings. This is particularly true when deaf clients have limited English proficiency, which often requires interpreters to use expansion techniques in order to render messages successfully. To examine how diagnostics may be affected by interpretation, Brauer (1993), Montoya et al. (2001), and Steinberg, Lipton, Eckhardt, Goldstein, and Sullivan (1998) translated two widely used psychological screening instruments into American Sign Language (ASL). The Minnesota Multiphasic Personality Inventory (MMPI) and the Diagnostic Interview Schedule-IV (DIS-IV) were selected for translation, and data from the three studies are presented and discussed. Their implications in terms of the expectations and stresses placed on interpreters are described within a framework of demand and control theory. Finally, sections of the Code of Ethics of the Registry of Interpreters for the Deaf (RID) are examined relative to both the issue of confidentiality and what the interpreter's contribution should be in mental health settings. PMID- 11865574 TI - Successful students who are deaf in general education settings. AB - The study identified successful students who were deaf and were receiving most of their educational services in general education settings, in order to examine factors contributing to their success. Teachers in a western state were asked to nominate students who were deaf who were in the upper elementary through high school grades and were receiving most of their educational services in general education classrooms. Qualitative procedures were used to gather information on 20 successful students who were deaf. Inquiry focused on observation of the students in general education settings and interviews to gather perceptions of (a) the successful students themselves, (b) deaf education teachers, educational interpreters, and paraprofessional note takers serving these students, (c) general education teachers working with these students, and (d) parents. The students' primary communication modes were closely divided between sign language and spoken English; communication mode did not seem to be a salient factor in success. Results of the interviews with each group, a summary of observations, and themes that emerged across groups are provided. PMID- 11865576 TI - [Morphologic and functional assessment of renal artery stenosis: use of combined MR angiography and MR renography--preliminary report]. AB - PURPOSE: To assess the possibility of use of multiphase magnetic resonance angiography for simultaneous acquiring of angiographic images and curves of contrast enhancement of renal cortex, medulla and pyelocalyceal system and evaluation of quality of obtained images. MATERIAL AND METHODS: Twenty-four patients suspected of having renal artery stenosis underwent power-injection of 30 ml of Gd-DTPA (3 ml/sec). From 10 to 360 seconds after the beginning of the injection, 3D MRA sequence was performed multiple times. Acquisition time of single phase of MR examination was 7.5-8 sec (TR = 5 ms, TE = 1:6 ms, single 7 cm thick slab with 35 partitions, 164 x 512 matrix). First three phases were used to obtain angiographic images. All phases were used to obtain curves of renal cortical, medullary and pyelocaliceal enhancement. Two readers evaluated quality of MRA images, as well as, quality of enhancement curves. RESULTS: 21 of 24 MRA examinations were of good, 2 of fair and none of poor quality. Quality of enhancement curves was good in 22 cases. It was suboptimal in 2 cases because of irregular breath-holding. Maximum number of acquisitions per minute was 4-5. Eight accessory, 2 obstructed and 9 stenosed renal arteries were visualized. Renographic curves were abnormal in 8 patients. CONCLUSIONS: MRA sequence with short acquisition time enables simultaneous acquisition of angiographic images and renographic curves of good quality. With further reduction in acquisition time it may be possible to obtain more points on MR renographic curves. PMID- 11865575 TI - [Renin-angiotensin system genes in chronic glomerulonephritis]. AB - Glomerulonephritis is a group of diseases with complex etiology, pathogenesis, morphological features and clinical course. The renin-angiotensin system genes are important group of candidate genes involved in pathogenesis of chronic renal diseases. The purpose of our study was to analyze the association of genetic polymorphisms of these genes with glomerular kidney diseases. The study population consisted of 52 patients with immunological glomerular kidney diseases and 50 hemodialyzed patients with end-stage renal failure with glomerulonephritis as primary disease. The control group consisted of 200 healthy subjects. By means of the polymerase chain reaction (PCR) the following polymorphisms were evaluated: insertion/deletion (I/D) polymorphism in intron 16 of the angiotensin converting enzyme gene (ACE), M235T polymorphism of the angiotensinogen gene (AGT) and A1166C polymorphism of the angiotensin II type 1 receptor gene (AT1R). No significant association was found between the ACE allele and genotype frequencies and the disease. The allele frequency of the M235T polymorphism was different from that observed in the control group, but differences in the genotype distribution were not statistically significant. The CC genotype of the AT1R gene polymorphism was significantly more frequent in patients than controls. This suggests an increased susceptibility to renal diseases in individuals carrying the CC genotype. This relationship is not associated with hypertension. Our results suggest that in the Polish population the AT1R gene polymorphism might be associated with increased susceptibility to chronic renal diseases. PMID- 11865577 TI - [Microvascular complications in pancreatic diabetes]. AB - Diabetes develops in more than half of the patients with chronic alcoholic pancreatitis (CAP), mostly due to increasing insulin deficiency. In this regard CAP-related diabetes (CAP-DM) is similar to the type 1 diabetes. Data on microvascular complications in CAP-DM are scarce. The aim of the study was the analysis of microvascular complications frequency in relation to metabolic control in comparison with type 1 diabetes mellitus. The study subjects were 50 patients divided into two groups: group 1-25 patients with CAP-DM (15 men, 10 women, mean age 44.6 +/- 8.4 yrs, duration of diabetes 3.7 +/- 2.1 yrs, body mass index (BMI) 22.4 +/- 2.9 kg/m2, duration of CAP 7.0 +/- 3.5 years), and group 2 25 well-matched type 1 diabetes patients (14 men, 11 women, mean age 42.3 +/- 7.6 yrs, duration of diabetes 4.1 +/- 2.8 yrs, BMI 24.0 +/- +/- 2.5 kg/m2). CAP was diagnosed on the basis of clinical examination, ultrasound and computed tomography scans, and in some cases upon the results of endoscopic retrograde pancreatography. Fasting plasma glucose, glycated hemoglobin (HbA1c), total serum cholesterol, triglycerides, urea and creatinine concentrations were measured. Fundoscopy was performed in all the subjects, in addition fluorescein examination was conducted in 15 and 18 patients from groups 1 as 2 respectively. Fasting plasma glucose, HbA1c level and insulin requirement were significantly lower in CAP-DM patients than in type 1 diabetes subjects (133 +/- 48 vs 174 +/- 59 mg/dl, p < 0.01; 8.3 +/- 2.0 vs 9.8 +/- 1.1%, p < 0.01; 36 +/- 15 vs 57 +/- 11 IU/day, p < 0.001 respectively). However, the prevalence of background retinopathy (group 1 13/25, group 2-11/25), and microalbuminuria (group 1-14/25, group 2-13/25) was similar in both groups. No statistically significant differences were found between CAP-DM and type 1 diabetic patients in regard to blood lipids, triglycerides, urea and creatinine concentrations. We conclude that microvascular complications may be encountered in pancreatic diabetes as often as in type 1 diabetes. Therefore this particular type of secondary diabetes should be regarded by no means as a "milder" type of the disease. PMID- 11865578 TI - [Effect of digoxin on atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP) and cyclic 3', 5'-guanosine monophosphate (cGMP) in patients with chronic congestive heart failure]. AB - Neurohumoral factors play important role in the pathogenesis of congestive heart failure (CHF) and digoxin (dig) is one of the most frequently used drugs in this condition. The aim of this study was to assess the effects of dig on atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP) and their "second messenger" cyclic 3',5'-guanosine monophosphate (cGMP). MATERIAL AND METHOD: The study group consisted of 25 patients (pts) aged 25-81 with CHF (NYHA II/III), 13 women and 12 men. Control group consisted of 10 healthy volunteers, 2 women and 8 men. The blood samples for evaluation of ANP, BNP and cGMP plasma level was taken at baseline conditions and 3 hours (h) after intravenous injection of 0.25 mg of dig. From the next day 0.25 mg dig was administrated orally for 6 days and plasma levels of ANP, BNP and cGMP were assessed on the 3rd and 6th day of treatment. Medium serum dig concentration on the 6th day was 0.98 ng/mL. RESULTS: The baseline ANP, BNP and cGMP plasma level was significantly higher in pts with CHD than in control group (ANP 144.4 vs 98.8 pg/ml p < 0.001; BNP 130.0 vs 97.2 pg/ml p < 0.001; cGMP 1.44 vs 0.86 pg/ml p < 0.001). In pts with CHF there was a significant increase ANP, BNP and cGMP plasma level 3 h after dig intravenous injection (ANP 205.2 vs 144.4 pg/ml p < 0.01; BNP 227.1 vs 130.0 pg/ml p < 0.01; cGMP 1.84 vs 1.44 pg/ml p < 0.01). The ANP and BNP plasma level after 3 days of oral dig was still significantly increased (ANP 171.1 vs 144.4 pg/ml p < 0.05; BNP 223.7 vs 130.0 pg/ml p < 0.01). ANP, BNP and cGMP plasma level was higher in pts with CHF also after 6 days of oral dig, but the difference was statistically significant. After 6 days of digoxin treatment there was a significant increase of ejection fraction (p < 0.005), with reduction of end-diastolic diameter of left ventricle (p < 0.05) and diameter of left atrium (p < 0.01). ANP at baseline correlated positively with baseline cGMP (r = 0.702 p < 0.05). On the 6th day BNP correlated positively with cGMP (r = 0.628 p < 0.05). 3 h after dig intravenous injection ANP correlated positively with BNP (r = 0.881 p < 0.05), but on the 3rd day of oral dig ANP correlated negatively with BNP (r = -0.536 p < 0.05). On the 6th day of oral dig end-diastolic diameter of left ventricle correlated negatively with BNP (r = -0.483 p < 0.05) and cGMP (r = -0.824 p < 0.05). CONCLUSIONS: 1. In pts with CHF a single intravenous digoxin injection increases ANP, BNP and cGMP plasma level. 2. Oral digoxin administration supports this beneficial neurohumoral effect and improves hemodynamic parameters of left ventricle as well as reduces left atrium diameter. PMID- 11865579 TI - [Is application of electrocardiographic exercise test always usefull in the diagnosis of coronary artery disease? Advantages and limitations of this method]. AB - To determine the diagnostic value of the electrocardiographic exercise testing (EET) in 551 patients with chest pain regarded as definite or probable stable angina pectoris (CAD), results of performed EET were compared with coronary angiography. All patients underwent exercise testing according to the Bruce protocol. The criterion for a positive exercise ST-segment response was > or = 1 mm of horizontal or down sloping depression 80 msec after J-point. The indications for cardiac catheterization in each patient were determined at the discretion of the attending physician. Clinically important coronary artery disease was defined as > 50 per cent narrowing of the diameter of at least one major vessel or > or = 50 per cent of the left main coronary artery. RESULTS: The sensitivity and specificity of EET for detection of CAD were for the entire group, in women and men respectively: 93%, 91%, 94% and 21%, 16%, 27%. CONCLUSION: 1. Indications for EET should be based on prior probability of coronary artery disease. 2. Application of higher than conventional ST depression criteria (> or = 2 mm) lowers sensitivity but increases specificity of EET. 3. Variables determining false positive results are as follows: age, sex (female), low probability of CAD, ST-segment depression in leads: II, III, aVF and mitral valve prolapse. 4. Variables determining false negative results are as follows: high probability of CAD, sex (male) and one vessel disease. PMID- 11865580 TI - [Deep hypotension with transient oliguria and severe heart failure in course of acute intentional poisoning with amlodipine]. AB - A case of acute poisoning with amlodipine with deep hypotension, transient oliguria and clinical signs of acute heart failure was described. A woman of 23 years swallowed intentionally 60 tablets of amlodipine (600 mg). After eleven hours of ingestion she was admitted to Warsaw Poison Control Centre. She was in severe clinical condition; tachycardia and deep hypotension were the prominent signs of poisoning. There was not CNS depression. Intensive treatment with i.v. catecholamines (dopamine, norepinephrine), crystalloids (with continuous control of central venous pressure), and i.v. calcium salts (with control of plasma calcium concentration) was started immediately. The patient did not improve but got worse. Acute heart failure developed, especially of left ventricle, so i.v. crystalloids were stopped and dubutamin, morphine, nitroglycerin and glucagon were introduced. Because of oliguria and insufficient effect of high doses of furosemide four-hours hemodiafiltration was set in. The patient's condition slowly improve after third and forth day of hospitalization. The systolic blood pressure rose, heart work was really better and on sixth day--the stabilization of diastolic blood pressure was definitely achieved. The patient was discharge in good condition with heart ejection fraction of 65% measured echocardiographically. PMID- 11865581 TI - [Pseudo-Gitelman's syndrome as consequence of loop diuretics abuse]. AB - We describe a case of 40-year old patient with chronic, resistant to treatment hypokalaemia. Differential diagnosis of renal potassium loss among Gitelman's syndrome, Bartter's syndrome and loop diuretic abuse was made. PMID- 11865583 TI - [A role of inflammatory and autoimmunizing factors in the etiology of MALT lymphoma]. PMID- 11865582 TI - [Induction of the immune tolerance--clinical trials]. PMID- 11865584 TI - [Enzymes conjugated with glutathione and their importance in medicine]. PMID- 11865585 TI - [Treadmill test for the assessment walking capacity of the patients with peripheral arterial occlusive disease]. PMID- 11865587 TI - [Adverse effects of parenteral administration of antisense oligonucleotides]. AB - To characterize the toxicity of phosphorothioate antisense oligodeoxynucleotides ([S]ODNs) in vivo, the mice received intravenously 26-mer bcr-abl antisense oligodeoxynucleotides (1 mg/mice/day) for 9 consecutive days. The organs and tissues were removed on the indicated days (+1, +7, +30) after the treatment. Our investigation revealed middle elevation of aminotransferases activity, lactate dehydrogenase level, total protein level and globulin level, decrease of glucose, albumin and blood urea nitrogen level in the peripheral blood. The mild anaemia and thrombocytopenia were observed too. The most significant treatment-related findings in the antisense treated mice were splenomegaly, reactive hepatitis and atrocytosis of kidney. These findings together with previous results demonstrate little and temporary toxicity effects mainly in organs known from cumulating of [S]ODNs. PMID- 11865588 TI - [Assessment of myocardial function at infarct zone after PTCA infarct-related artery--a study with tissue Doppler echocardiography]. AB - Angioplasty of an infarct-related artery (IRA) performed several weeks or months after myocardial infarction (MI) may improve myocardial function. It is still unclear though, how soon wall function is restored following the procedure. This study was designed to assess quantitatively changes of regional left ventricular function after PTCA of IRA by means of tissue Doppler echocardiography (TDE). Thirty nine patients (30 male, mean age 53.4 +/- 8.4 yrs) who had MI 13 +/- 6 weeks earlier were qualified for IRA angioplasty on the basis of dobutamine stress echocardiography (DSE) if a viable myocardium was demonstrated. Regional wall function at infarct zone was assessed by TDE one day before PTCA (exam 1), 1 3 days (exam 2) and 28-30 days (exam 3) after successful angioplasty. Myocardial velocities and time--derived TDE intervals were calculated both in systole (systolic peak velocity--S, pre-ejection period--PEP, contraction time--CT), and diastole (E and A velocity waves, E/A ratio, isovolumic relaxation time--IVRT, rapid filling time--RFT, atrial filling time--AFT). All parameters were measured in longitudinal direction (annulus, basal and medial segments) of posterior (20 pts), anterior (17 pts) and lateral (2 pts) walls. S wave velocity increased between exam 1 and 2 (4.9 +/- 1.2 cm/s vs 5.6 +/- 1.3 cm/s, p < 0.02), whereas E wave decreased between examinations. Pre-PTCA E/A ratio was significantly lower than in exam 2 and 3. PEP decreased between exam 1 and 2 (96 +/- 23 ms vs 84 +/- 16 ms, p < 0.01). Significant correlation was found in IVRT and RFT prior and immediately after PTCA (103 +/- 21 ms vs 87 +/- 20 ms, p < 0.001, 151 +/- 39 ms, vs 170 +/- 30 ms p < 0.01 respectively). CONCLUSIONS: Patients after infarction with persisted viable myocardium may benefit from late angioplasty of IRA. In regional wall function assessment TDE seems to be more sensitive technique than visual wall motion analysis alone. TDE parameters demonstrated a rapid initial improvement. Changes of myocardial velocities (S, E) and time--derived TDE intervals (IVRT, PEP, RFT) are sensitive markers of restored myocardial function. PMID- 11865589 TI - [Evaluation of expression of polymorphonuclear neutrophil surface receptors in patients with type 1 diabetes]. AB - Polymorphonuclear neutrophils (PMN) play an important role in the pathogenesis of diabetic microvascular complications. Stimulation of these cells is associated with the appearance of specific receptors on their surface. The aim of the study was to evaluate the expression of the receptors specific for PMN: CD 11b, CD 18. The study was performed in a group of 23 type 1 diabetic patients, aged from 19 to 47 years (mean 30.7 +/- 8.6 years), including 15 females and 8 males (mean diabetes duration time 13.7 +/- 7.5 years; mean HbA1c 7.9 +/- 2.5%). The expression of PMN surface receptors was measured by flow cytometry using a ORTHO DIAGNOSTIC SYSTEM cytofluorimeter. The results were presented as a PMN percentage indicating expression of CD 11b and CD 18. In comparison to healthy controls, there was a significant increase in the number of PMN, both with CD 11b and CD 18 receptors present--(CD 11b: 93.2 +/- 3.4 vs 98.0 +/- 1.9% p < 0.05), (CD 18: 98.5 +/- 0.47 vs 99.4 +/- 0.7%, p < 0.05). CONCLUSION: In patients with type 1 diabetes, PMNs demonstrate a pronounced expression of surface receptors which may indicate an enhanced activity of these cells. The increase of expression of surface receptors is independent of diabetes duration time and HbA1c. PMID- 11865590 TI - [Vitamin C and uric acid concentrations in plasma and gastric mucosa in patients regularly consuming vegetables, fruits and fruit juices]. AB - Vitamin C (Asc) as a reactive oxygen species scavenger inhibits detrimental oxidation reactions and counteracts formation of mutagenic compounds which damage DNA in the gastric and duodenal mucosa. It has been considered as an important factor in reducing risk of gastric carcinoma. However, the role of uric acid (UA) as an antioxidant factor and its influence on gastric mucosa has not been so well investigated. The aim of the study was to evaluate the relationship between vegetable, fruit and fruit juices intake and water soluble antioxidant (Asc and UA) concentrations in plasma and gastric mucosa. The study also assessed the prevalence and intensity of inflammatory changes in mucosa and Helicobacter pylori infection. 34 patients participated in the study. Asc and UA concentrations in plasma and gastric mucosa samples were examined with the use of liquid chromatography (HPLC) method. Histopathological examinations of gastric mucosa were also performed. Higher concentrations of Asc and UA in plasma and less frequent inflammatory changes of gastric mucosa were found in patients regularly consuming vegetables and fruits. It may suggest the protective effect of these compounds on mucosa. No significant relationship was found between H. pylori infection and antioxidant concentrations in plasma and gastric tissue in patients with chronic gastritis. CONCLUSIONS: Diet rich in fruits, vegetables and fruit juices helps in reducing prevalence of inflammatory changes in gastric mucosa. It can be associated with higher Asc and UA concentrations in plasma and higher UA concentrations in gastric mucosa. Lower Asc and UA concentrations in patients with gastritis have no significant relationship with H. pylori infection. PMID- 11865591 TI - [Uremic neuropathy is more frequent in male patients]. AB - The aim of this study was to analyse the relationship between clinical and electrophysiological features of somatic and autonomic neuropathy and gender of 51 patients on chronic hemodialysis. Apart from basic neurological examination, conduction velocities in peripheral nerves were determined, and the function of the autonomic nervous system was assessed with the help of two tests: R-R interval variation (RRIV) and sympathetic skin response (SSR). The incidence and intensity of clinical and electrophysiological signs of sensomotor neuropathy were statistically significantly more prevalent among male patients, whereas the symptoms suggesting autonomic involvement were more frequent in women. Results of parasympathetic electrophysiological tests were similar in both groups, but abnormal SSR results prevailed in male patients. PMID- 11865592 TI - [An oligosymptomatic case of pheochromocytoma]. AB - A case of 34-year old female with incidentally diagnosed adrenal tumour is discussed. The patient complained only of mild headaches and heart palpitations and was not previously treated for hypertension. A diagnosis of pheochromocytoma was made. The diagnostic controversies arose because of subclinical course of the disease, slightly elevated biochemical markers of pheochromocytoma (catecholemines urinary excretion) and non-characteristic result of glucagon stimulation test results. The diagnosis was confirmed by histologic examination of tumour tissue. Presented case indicates the need for thorough clinical and hormonal evaluation of patients with incidentaloma (particularly, when adrenal tumour diameter is larger than 3 cm) to avoid serious complication of surgery treatment in case of misdiagnosis. PMID- 11865593 TI - [Chylomicronemia syndrome in 45 years old man]. AB - The case of chylomicronemia syndrome in 45 year old man, previously misdiagnosed as hypercholesterolemia is described. Secondary causes of hyperlipoproteinemia were excluded. No symptoms, characteristic of familial lipoprotein lipase deficiency were observed. We concluded that the diagnosis of hyperlipoproteinemia has to be based on determination of all plasma lipids concentrations (total cholesterol, triglycerides, LDL cholesterol, HDL cholesterol). Otherwise a false diagnosis is quite possible. In rare cases additional laboratory tests are needed. PMID- 11865594 TI - [The role of alpha-1-antitrypsin phenotypes in the pathogenesis or rheumatic diseases]. PMID- 11865595 TI - [The role of costimulatory molecules in immune responses in acute leukemia]. PMID- 11865596 TI - [Autonomic neuropathy in uremic patients]. PMID- 11865597 TI - [Kawasaki disease]. PMID- 11865598 TI - [Osteoimmunology: new area of research on the associations between the immune and bone systems]. PMID- 11865599 TI - New way to detect cervical cancer. PMID- 11865600 TI - Nursing project takes aim at pediatric cancer. PMID- 11865601 TI - Health technology assessment. PMID- 11865602 TI - Men in nursing. An untapped resource. PMID- 11865603 TI - Patient-focused care. A dialogue with your patient. PMID- 11865604 TI - Midwifery what nurses know and think. PMID- 11865605 TI - Evening shift. PMID- 11865606 TI - Nursing research. PMID- 11865607 TI - The abuse stops here. PMID- 11865608 TI - AIDS prevention on the streets. PMID- 11865609 TI - A perioperative community link with families. PMID- 11865610 TI - Taking the leap to gerontological certification. PMID- 11865611 TI - Don't tolerate abuse. PMID- 11865613 TI - Fruitful questioning. PMID- 11865612 TI - A police officer's perspective. PMID- 11865614 TI - Nursing policy. Making the talk matter. Interview by Marla Fletcher. PMID- 11865615 TI - Safe babies. PMID- 11865616 TI - Female nurses in post-secondary education. PMID- 11865617 TI - Health & safety. Sleep and shiftwork. II. PMID- 11865619 TI - [Barrett's esophageal cancer]. AB - Barrett's esophageal cancer is defined as carcinoma developing in Barrett's esophagus. The esophagogastric junction is located at the distal end of a network of fine longitudinal vessels, and the columnar epithelium existing above it is Barrett's mucosa. Barrett's mucosa, especially specialized columnar epithelium is considered as precancerous lesion, and malignant potential is examined in various ways. For the surveillance of malignant lesions from Barrett's esophagus, periodic endoscopic examination is necessary with chromoendoscopy or magnifying endoscopy. Treatment strategies are EMR and other endoscopic treatment for mucosal cancer, and surgical treatment for submucosal and advanced cancer. Several surgical modalities are employed depending on the stage of cancerous progression, the location of the cancer in Barrett's esophagus, and the length of Barrett's esophagus. There remain many unexplained problems in Barrett's esophagus and Barrett's cancer. PMID- 11865618 TI - Changes in approach. PMID- 11865620 TI - [Present role and future prospect of superselective intra-arterial infusion chemotherapy for head and neck cancer]. AB - Patients with head and neck squamous cell cancers (HNSC) of stage III and IV have a poor outcome, often losing important functions such as swallowing and speech despite combined surgery and radiotherapy. Two-thirds of such patients die of local recurrence and disseminated metastasis in spite of salvage therapy. Conventional adjuvant chemotherapy is often difficult for HNSC patients, since many of them suffer from associated chronic oral, respiratory and gastrointestinal diseases due to tobacco and alcohol in addition to having poor nutritional and oral hygienic conditions. Ninety-seven patients with advanced head and neck cancers were treated by superselective intra-arterial chemotherapy using CDDP and sodium thiosulfate (STS) from 1995 to 2000. Once a week, 100-150 mg/m2 of CDDP were administered superselectively at 5 mg/min through each artery feeding the tumor. During the infusion of CDDP, STS at a dose of two hundred fold that of CDDP was injected through a catheter placed in the subclavian vein. The complete and partial response rates were 72% and 25%, respectively, with 100% preservation of the larynx and 90% preservation of the eyeball in all involved cases. We could suppress mucositis of normal tissue and chemotoxicities leading to conditions such as renal and hematological dysfunction. Though our method currently has disadvantages such as the risk of cerebral infarction, the fact that chemotherapy must always be done under fluoroscopy, and the occurrence of distant metastasis, there are good prospects for the future and we are now working toward solving the present problems. PMID- 11865621 TI - [Intra-arterial infusion chemotherapy for breast cancer]. AB - Intra-arterial infusion chemotherapy has a significant effect in down-staging locally advanced breast cancer by providing a high dose intensity. It is also used for the treatment of liver metastasis. A better response rate and lower incidence of adverse effects are reported when patients are treated with intra arterial infusion chemotherapy than with systemic chemotherapy using the same dose of the same drugs. Recent advances in devices such as access ports and portable infusion pumps make it possible to perform intra-arterial infusion chemotherapy repeatedly and safely. However, it remains uncertain whether or not intra-arterial infusion chemotherapy can improve the prognosis of cancer patients because of the lack of data from phase III trials. Accordingly, further studies including combined use of systemic chemotherapy are mandatory to the control of micrometastases outside the target organ. PMID- 11865622 TI - [Evidence for and practical use of arterial infusion chemotherapy for liver metastases]. AB - Previous clinical trials have shown a significantly higher tumor response to arterial infusion chemotherapy for liver metastases compared with systemic chemotherapy, but failed to show the survival impact. Thus, there has been no evidence to justify the use of this therapy for liver metastases. On the other hand, the results of previous clinical trials cannot be used to decide the role of this therapy, because the trials were performed under incomplete situations in which there was no consideration of the unreliability of imaging diagnoses and the importance of techniques. At present, therefore, we should use this therapy based on deep understanding and thought toward previous data and patients' conditions. Clinical trials involving considerations for this special therapy are required in order to decide the role of this therapy. PMID- 11865623 TI - [Recent advances in intraarterial chemotherapy in gynecologic malignancy]. AB - Anti-cancer chemotherapy (CTX) agents are mainly given intravenously (i.v.) in patients with gynecologic malignancy. The reasons intraarterial chemotherapy (IA CTX) are not more commonly used are the following: 1) cervical, corpus, and ovarian carcinoma have shown an appreciable response to i.v. cisplatin-based chemotherapy, 2) locally advanced carcinoma, suitable for IA-CTX, is a potentially systemic disease, 3) IA-CTX often requires a special technique, 4) IA CTX has not been a major concern for the Gynecologic Oncology Group (GOG) in the United States, and 5) the very small amount of medical evidence supporting IA-CTX makes cancer patients hesitate to give informed consent prior to receiving IA CTX. The major concern of gynecologic oncologists has already moved away from the administration route of agents to new agents themselves including paclitaxel, docetaxel, and CPT-11. Looking again at the clinical data of IA-CTX as a neoadjuvant CTX for advanced cervical carcinoma reported in the 1990s, the response rate seems to be superior to that achieved by i.v.-CTX. Thus, we must now reappraise the IA-CTX for gynecologic malignancy. PMID- 11865624 TI - [Intra-arterial chemotherapy in urological cancer]. AB - We herein describe the present status of intra-arterial chemotherapy for patients with renal cell carcinoma and bladder cancer, and discuss the prospect of intra arterial chemotherapy in these urological diseases. The combination of immunotherapy and intra-arterial chemoembolization with microcapsules may prolong the survival of patents with unresectable tumors. However, the role of intra arterial chemotherapy in renal cell carcinoma is limited. For bladder cancer, several modifications to increase the effect of intra-arterial chemotherapy have been documented. The combination of intra-arterial chemotherapy and radiotherapy resulted in a complete response rate of 84-91%, supporting the possibility of bladder preservation for patients with invasive bladder cancer. To establish the effectiveness of intra-arterial chemotherapy, large scaled prospective randomized studies are necessary. PMID- 11865625 TI - [Globalization of anti-cancer therapies]. AB - Based on short reviews of lung, gastric, colorectal, prostate, breast and ovarian cancer, there remain significant differences between Japan and the West in the therapeutic regimen for most cancers. Some of the differences are due to differences in stage of disease at diagnosis or historical factors affecting availability of products. In both Japan and the West, there are initiatives to prepare treatment guidelines based on published data. For Japan this initiative is limited by the lack of Japanese clinical trial data or even safety data. When guidelines are prepared from international data, many of the products have limited indications in Japan and therefore not reimbursed. Availability of the most appropriate therapies to Japanese patients will depend on a facilitation of clinical trials in both primary and additional indications. However, the experience in other countries is that, even where data and registration approval are available, guidelines are hard to agree and are not uniformly accepted by prescribers. The ICH E5 guideline on the use of bridging studies to interpolate Western data to Japanese regulatory dossiers provides an opportunity to accelerate availability of new medicines to Japanese prescribers and patients. The use of bridging studies has so far been limited for anti-cancer therapies. Where relevant pharmacodynamic endpoints can be measured, (e.g. aromatase inhibition) there can increase confidence in bridging. The newer types of agent which act to stabilise disease rather than tumour shrinkage present a special problem. In some cases surrogate markers can be valuable but in each case they need to be validated. As globalization continues, an alternative approach is to include a significant cohort of Japanese patients in Japanese patients but this depends on sufficient similarity in the patient population and background therapy. The most significant limitation to either large outcome studies in Japan or for Japanese centers to join international trials has been the environment for conduct of clinical trials. There have been some recent improvements and further progress is expected so that Japanese doctors can play a full role in the evaluation of new therapies. PMID- 11865626 TI - [Apoptosis as a measure of chemosensitivity to anti-cancer drugs in gynecological cancer]. AB - OBJECTIVE: To examine apoptosis as a measure of chemosensitivity to anti-cancer drugs in gynecological cancer xenografted into nude mice. METHODS: The TUNEL method was used to investigate the induction of apoptosis after exposure to anti cancer drugs in thirteen gynecological cancers xenografted into nude mice. Moreover, in vivo sensitivities of gynecologic cancer to anti-cancer drugs were determined according to the protocol developed by Battle Laboratories, Columbus, Ohio USA. Both methods were examined for correlation. RESULTS: The apoptosis index by the TUNEL method showed a correlation with the findings using the protocol developed by Battle Laboratories, Columbus, Ohio USA. CONCLUSIONS: Our findings indicate the ability to determine the measure of chemosensitivity by the apoptosis index. PMID- 11865627 TI - [Weekly paclitaxel therapy for metastatic breast cancer]. AB - We treated 12 patients with metastatic breast cancer with weekly paclitaxel therapy. Paclitaxel was administrated by 1 hour infusion at a dose of 80 mg/m2 after short premedication every week on an outpatient basis. Administration was continued for 3 weeks followed by 1 week rest. All patients had received prior metastatic chemotherapy, and prior anthracycline therapy was done in 66.7% of the patients. Partial responses were observed in 66.7% of the patients and progressive disease in 33.3%. The response rate was 66.7%. Responses were observed in 62.5% of the patients with prior anthracycline therapy. Grade 3/4 leukopenia and neutropenia occurred in 25% of the patients, respectively, and no grade 3/4 peripheral neuropathy was observed. Dyspnea occurred in 25% of the patients and was grade 3 in 16.7%. Dyspnea is thought to be one of the adverse events requiring caution with weekly paclitaxel administration. Weekly paclitaxel therapy is effective and well tolerated in patients with metastatic breast cancer. PMID- 11865628 TI - [Weekly administration of low-dose paclitaxel for advanced or metastatic breast cancer]. AB - The activity and toxicity of a weekly infusion of low-dose paclitaxel was studied. Twelve patients with metastatic or advanced breast cancer received paclitaxel (80 mg/m2 over 1 h) every week. Administration was continued for 6 weeks with two weeks rest until disease progression or limiting toxicity. Dexamethasone 20 mg, diphenhydramine 50 mg, and ranitidine 50 mg were given prior to each dose of paclitaxel. Six patients had received prior standard CMF therapy, and four patients had received CMF and docetaxel therapy. Two patients had not received prior therapy. The overall response rate was 58% with 17% complete responses and 42% partial responses. Responses were observed in both patients without prior therapy, and in five of 10 (50%) with prior therapy. Grade 3/4 neutropenia occurred in one patient; febrile neutropenia was not observed. There was no neuropathy or hypersensitivity. Weekly paclitaxel is active and well tolerated in patients with metastatic or advanced breast cancer. This schedule allows a high cumulative dose of paclitaxel without major myelo- or neurotoxicity. This weekly regimen deserves further exploration. PMID- 11865629 TI - [Preliminary clinical evaluation of continuous infusion of 5-FU and low-dose Cisplatin (LFP) therapy alone and combined with radiation therapy for treatment of advanced or recurrent esophageal cancer]. AB - We evaluated the clinical effect of 5-FU and low-dose Cisplatin (LFP) therapy alone and LFP therapy combined with radiation therapy in patients with advanced or recurrent esophageal cancer. From March 1995 to September 2000, 11 patients with inoperable esophageal cancer, 8 patients with adjuvant chemotherapy post operation, and 14 patients with recurrent esophageal cancer were treated with LFP therapy. 5-FU (160 mg/m2/day) was continuously infused over 24 hours, and CDDP (3 7 mg/m2/day) was infused for 30 minutes. The administration schedule consisted of 5-FU for 7 consecutive days and CDDP for 5 days followed by a 2-day rest, each for four weeks. We combined radiation therapy for the patients with all lesions that could be included in the radiation field. Of 30 patients with measurable lesions the response rates of LFP therapy alone and LFP therapy combined with radiation therapy were 33% and 60%, respectively. Toxicity over grade 3 appeared in 3 of 15 patients with LFP therapy combined with radiation therapy. There was no significant difference between LFP therapy alone and LFP therapy combined with radiation therapy with regard to survival rate of inoperable and recurrent esophageal cancer. In conclusion, LFP therapy alone may be effective for esophageal cancer. PMID- 11865630 TI - [The clinical effect of TS-1 in advanced and recurrent gastric cancer with peritoneal dissemination]. AB - Eighteen patients with far advanced and recurrent gastric cancer with peritoneal dissemination were treated with a novel oral anticancer drug, TS-1, and assessed according to clinical effect. TS-1 was administered at a dose of 80-120 mg/day. One course consisted of consecutive administration of TS-1 for 28 days followed by 14 days rest. The 1- and 2-year survival rates and median survival time after administration of TS-1 were 63.2%, 23.7% and 437 days, respectively. Eight patients (44.4%) survived for 1 year or more. Adverse reactions consisted of reduction in hemoglobin level and hyperbilirubinemia at grades 3 and 4, which were observed in 3 patients and 1 patient, respectively. TS-1 is a promising drug for gastric cancer with peritoneal dissemination. PMID- 11865631 TI - [Dihydropyrimidine dehydrogenase expression in gastric cancer using anti-human dihydropyrimidine dehydrogenase monoclonal antibody]. AB - The rate-limiting enzyme to catabolize 5-fluorouracil (5-FU) is dihydropyrimidine dehydrogenase (DPD), which expression in cancerous tissue is reported to have a relation with anti-tumor effect for 5-FU. In this study, we evaluated the immunohistochemical expression in gastric cancer using two different kinds of anti-human DPD antibody, KM1915 and KM1919. However, recognition of both antibody was mimetic in human gastric cancer, and strong expression of DPD was observed in the interstitial tissue when using KM1919. There is a positive relationship between immunohistochemical expression by KM1915 and mRNA expression in human gastric cancer. Immunohistochemical study with KM1915 might be a clinically useful method to evaluate the expression of DPD in paraffin-embedded materials. PMID- 11865632 TI - [Styrene maleic acid neocarzinostatin-transcatheter embolization for hepatocellular carcinoma--third report]. AB - To evaluate the effect of styrene maleic neocarzinostatin-transcatheter arterial embolization (SMANCS-TAE), 40 patients with unresectable hepatocellular carcinoma (HCC) of hypervascular radiological feature, associated with liver cirrhosis (LC), 18 in clinical stage 2 and 20 in stage 3, were treated by SMANCS-TAE. SMANCS with Lipiodol and then gelatin sponge particles were injected into the artery branch supplying HCC using selective catheterization, and its effect was evaluated by computed tomography (CT) Grade. In patients with Grade III or less (Lipiodol accumulation < 99% in the entire tumor) after the first course of therapy, SMANCS-TAE or arterial injection of SMANCS-Lipiodol was performed once or twice more. Consequently, 32 of 40 patients (80%) obtained Grade IV (100% Lipiodol accumulation in the entire tumor) after from once to thrice (median, 1.6 courses). Grade IV was maintained in 26 of 32 patients, and non-recurrence was found 16 of 40 (40%) at the primary tumor to the time at last of follow up. Severe side effects were not noted except in 10 cases with narrowness of hepatic artery and cases of 2 biloma in patients undergoing therapy two or more times. The 1-, 2-, 3-, and 5-year survival rate was 85, 64, 35, and 26%, respectively. No significant difference was noted in the survival rate between clinical stage 2 and 3 liver cirrhosis (LC). But the survival rate of patients who continued to exhibit Grade IV at the primary tumor was significantly better than in those exhibiting Grade III or less (96, 68, 56, and 43% vs 64, 29, 0, and 0%, respectively; p < 0.01). In conclusion, the HCC patients, even those with decompensated LC, who obtained and maintained Grade IV after SMANCS-TAE could reduce the courses of treatment without severe side effects and survived longer. SMANCS-TAE might be useful for the good quality of life of HCC patients. PMID- 11865633 TI - [Clinical usefulness of ondansetron injection in patients receiving cancer chemotherapy]. AB - Before and after launch of 5-HT3, antagonist, necessary dose and duration of chemotherapy were compared between the patients who were confirmed to have undergone chemotherapy before the launch of 5-HT3 antagonist (retrospective group) and the ones currently using ondansetron (OND) for chemotherapy-induced emesis (prospective group). Clinical usefulness of OND was evaluated through survey on quality of life (QOL) to patients and questionnaires to physicians, nurses & patients. Necessary dose of chemotherapy was evaluated by investigating actual dose of cisplatin (CDDP). As the result, necessary dose of CDDP was confirmed to be different between retrospective and prospective groups. The influence on the actual CDDP dose was observed with or without use of G-CSF or by recommended dose of CDDP, while no influence by 5-HT3 antagonist was observed. For necessary duration of chemotherapy, significant difference was not observed between retrospective or prospective groups. On the other hand, actual CDDP dose or necessary duration of chemotherapy were confirmed to be greatly affected by chemotherapy-induced adverse events such as blood disorder (e.g. bone marrow suppression) or renal disorder, rather than chemotherapy-induced emesis. As the result of QOL survey to patients and other questionnaires to medical staff & patients, the fact of chemotherapy-induced emesis to lower the patient's QOL as well as the importance of emetic control was confirmed. It was also confirmed that the workload of nurses or other medical staff has lessened since the launch of 5-HT3 antagonists. PMID- 11865634 TI - [Clinical usefulness of ondansetron hydrochloride for nausea and vomiting during repeated courses of chemotherapy for malignant lymphoma--impact of prognosis announcement on anti-emetic effect and evaluation of patient perception of chemotherapy-associated adverse events]. AB - We evaluated the efficacy and safety of ondansetron hydrochloride (OND) on nausea and vomiting during repeated courses of CHOP or ACOMP-B therapy in patients with malignant lymphoma. The impact of the prognosis announcement on the anti-emetic effect and chemotherapy-associated adverse events was also investigated. Forty two subjects with malignant lymphoma who underwent CHOP or ACOMP-B therapy including cyclophosphamide 600 mg/m2 and adriamycin 40 mg/m2 were investigated for a maximum of 6 courses. For acute nausea and vomiting, ondansetron was injected intravenously before the start of chemotherapy on the first day of each course of chemotherapy. For delayed emesis, ondansetron was administered orally for 4 days from the following day. The efficacy on acute nausea and vomiting was found to be 95.0% (1st course), 95.0% (2nd course), 90.9% (3rd course), 88.2% (4th course), 92.3% (5th course) and 91.7% (6th course), respectively. A high efficacy of > or = 85% was also obtained for delayed nausea and vomiting on each day. Though the adverse event of elevated GPT value developed in one subject. It was mild and resolved. No difference in efficacy was seen with or without announcement of prognosis to patients. Following the investigation on antiemetic effect, patient perception of chemotherapy-induced adverse events was evaluated. The most common event was hair loss, followed by taste abnormality and numbness and hyposthesia of the tips of the fingers. The incidence of nausea and vomiting was the 4th and 5th most common, which are less frequent than in the report of Coates in 1983. In conclusion, ondansetron is considered clinically useful with stable anti-emetic effect on both acute and delayed nausea and vomiting over repeated courses of chemotherapy, without any significant safety problem. PMID- 11865635 TI - [Development of a Japanese version of the FLIE]. AB - Nausea and vomiting induced by chemotherapy have an impact on cancer patients' quality of life (QOL). The Functional Living Index-Emesis (FLIE), which is designed to assess the change in QOL from the influence of nausea and vomiting is rarely used in Japan, regardless of its utility, because it is written in English. We investigated the use by cancer patients with the main object of designing a reliable and valid Japanese version of the FLIE. We also verified the validity of a Japanese translation and improved part to design a highly precise Japanese version FLIE. Consequently, we found a correlation between the FLIE Japanese version and the QOL questionnaire Quality of Life Assessment of Cancer Patients Receiving Chemotherapy (QOL-ACPRC), which was the external standard. Furthermore, we improved the questionnaire to raise the rate of patient response, and improve reliability and validity. We think that this FLIE Japanese version will become useful in assessing the change in patient QOL due to the influence of nausea and vomiting. PMID- 11865636 TI - [A case of advanced gastric cancer successfully treated with TS-1]. AB - A patient with advanced gastric cancer complicated with liver and lymph node metastases was successfully treated with a novel oral anticancer drug, TS-1, TS-1 was administered at a dose of 100 mg/day. One course consisted of consecutive administration of TS-1 for 28 days and withdrawal for 14 days. At the end of 3 courses a partial response of the liver metastases was achieved. Although the patient has had complications with ascites collection due to hypoalbuminemia, he has been well without regrowth of any metastases for over 8 months. PMID- 11865637 TI - [Dormant chemotherapy by low-dose FP and low-dose UFT-E in recurrent gastric cancer with long-term survival--a case report]. AB - We experienced a case of recurrent gastric cancer with a long-term survival. A 64 year-old man was admitted to the hospital for advanced gastric cancer in the upper stomach. Abdominal CT scan revealed para-aortic lymph nodal metastases. The patient underwent total gastrectomy, distal pancreatectomy, splenectomy, left adrenectomy, and left nephrectomy with D4 lymph node dissection, in what was a curability B resection. Conclusive findings were t2 (ss), n4, H0, P0, M0, and stage IVb. One year after the operation, para-aortic lymph node recurrence was evaluated. The patient was treated with low-dose cisplatin-5-FU therapy, and a partial response was observed and continued for over 2 years with an administration of UFT-E (300 mg/day). He died of repeated aggravation of para aortic lymph node metastases 6 years and 2 months after the operation. We considered that the long-term survival of this patient was attributable to a 3 year tumor dormancy induced by low-dose cisplatin-5-FU therapy and administration of low-dose UFT. PMID- 11865638 TI - [Three cases of advanced gastric cancer treated by TS-1 in combination with low dose cisplatinum]. AB - TS-1, a DPD inhibitory fluoropyrimidine, is a novel oral formation of 5 fluorouracil (5-FU). In patients with advanced gastric cancer, the response rate was reportedly over 40%. We report three cases of advanced gastric cancer treated using TS-1 in combination with a low-dose of cisplatinum (CDDP) that well responded. Case 1: A 62-year-old women underwent total gastrectomy. Ten weeks later, she suffered intestinal obstruction due to peritoneal recurrence of gastric cancer. Eighty mg of TS-1 in combination with bi-weekly administration of CDDP (10 mg) improved her intestinal obstruction. Case 2: A 50-year-old man suffered peritoneal recurrence of gastric cancer. Computed tomography (CT) showed intestinal obstruction, ascites, and hydronephrosis. After 100 mg of TS-1 in combination with bi-weekly administration of CDDP (20 mg) for 1 year, CT showed almost complete improvement of peritonitis carcinomatosa. Case 3: A 58-year-old man, who suffered advanced gastric cancer with peritonitis carcinomatosa, was administrated 100 mg of TS-1 in combination with bi-weekly administration of CDDP (20 mg). After 2 months of administration, remarkable improvement was observed in the upper gastrointestinal series. Adverse reactions, which were grade 1 for stomatitis, were observed only in case 1. All three patients are alive (case 1 and 2 have survived more than one year) and therapy is continuing. In conclusion, combined chemotherapy of TS-1 and low-dose CDDP was effective and well tolerable for advanced gastric cancer patients. It was suggested that effective biochemical modulation might be achieved by these two drugs. PMID- 11865639 TI - [A case of advanced ovarian carcinosarcoma that responded remarkably to neoadjuvant chemotherapy of combined CPT-11 and CDDP]. AB - Carcinosarcoma of the ovary is a very rare and highly malignant neoplasm that accounts for less than 1% of ovarian neoplasms. Survival of patients with advanced stage cancer is poor and the best treatment is not clear. We report the case of a 60-year-old woman who had Stage IV advanced heterogeneous ovarian carcinosarcoma with lung and liver metastases. The lesions were considered surgically incurable, so she was placed on neoadjuvant chemotherapy of combination CPT-11 (60 mg/m2, day 1, 15) and CDDP (60 mg/m2, day 1). Tumor markers of CA125 and LDH decreased remarkably to the normal level after 3 and 4 courses of chemotherapy, respectively. After 7 courses of chemotherapy, the ovarian tumor was obviously reduced, and the lung and liver metastases had disappeared. The patient was then able to undergo surgery. The current case suggests that combination CPT-11 and CDDP is effective against advanced ovarian carcinosarcoma. PMID- 11865641 TI - [A case of long surviving advanced recurrent breast cancer with multiple bone metastases responding to treatment with 5'-DFUR combined with MPA]. AB - The patient was a 69-year-old woman who had undergone right standard radical mastectomy on August 8, 1991, and was treated with chemo- and hormonal therapy of ADM, UFT and TMA. Three years later she showed multiple bone metastases with elevation of CEA, and 5'-DFUR 1,200 mg/day and MPA 800 mg/day were administered. Two years later her CEA levels were decreased, 5'-DFUR was discontinued and MPA 1,200 mg/day only was continued. Two months later a side effect of MPA, her body weight gain, was observed, and the dosage of MPA was reduced from 1,200 mg/day to 800 mg/day. Then the side effect was resolved. Bone scintigraphy and MRI showed that bone metastatic lesions were reduced 6 years after 5'-DFUR and MPA therapy. It is suggested that this combination therapy may be useful for advanced recurrent breast cancer patients with multiple bone metastases. PMID- 11865640 TI - [A case of extraovarian primary peritoneal carcinoma successfully treated with weekly paclitaxel]. AB - We report the use of paclitaxel in the successful treatment of a 74-year-old patient with extraovarian primary peritoneal carcinoma and acute renal failure caused by intraperitoneal cisplatin. The histological diagnosis was papillary serous adenocarcinoma. Paclitaxel was given intravenously at 70 mg/m2 in a 1-hour infusion weekly for 6 consecutive weeks followed by 2-weeks rest (one cycle). Toxicities including neutropenia and neurotoxicity were mild. She showed a clinical response after three cycles, and then underwent secondary cytoreductive surgery after which she received one additional cycle of therapy. She enjoyed a favorable quality of life without evidence of disease for 16 months after the completion of this therapy. Weekly 1-hour paclitaxel (70 mg/m2) was well tolerated, yet was effective for extraovarian primary peritoneal carcinoma. PMID- 11865642 TI - [Reversible cardiomyopathy secondary to interferon-alpha in chronic myelogenous leukemia]. AB - Interferon-alpha (IFN-alpha) has been accepted as an effective agent in the treatment of chronic myelogenous leukemias (CML). Cardiac toxicity of IFN-alpha has rarely been reported in cases of CML. A 62-year-old woman with a two-year history of chronic myelogenous leukemia who had been treated with IFN-alpha (10 million U/3 days/week) given subcutaneously with oral hydroxycarbamide 500 mg, presented with chest pain, dyspnea and subconsciousness. Chest X-ray revealed cardiomegaly and congestion, and ultrasonography showed diffuse hypokinesis of the heart with decreased left ventricular ejection fraction (LVEF) 34%. She was diagnosed cardiomyopathy caused by IFN-alpha administration. She was treated with furosemide, dobutamine hydrochloride, milrinone and carperitide. The administration of IFN-alpha was terminated. LVEF was improved to 50% within one month from the onset of events, and the patient was discharged. We discuss herein the cardiomyopathy caused by IFN-alpha in CML. PMID- 11865644 TI - [Circadian chronotherapy with cisplatin and 5-fluorouracil in three patients with advanced gastric cancer]. PMID- 11865643 TI - [A case of advanced hypopharyngeal carcinoma successfully treated with superselective intra-arterial infusion of docetaxel]. AB - Docetaxel is an excellent agent with a high antitumor effect for advanced/recurrent head and neck cancer. A 67-year-old male with advanced hypopharyngeal cancer (T3N2bM1: Stage IV) underwent two courses of superselective intra-arterial infusion of docetaxel and intravenous administration of CDDP and 5 FU. Using a coaxial technique, a microcatheter was placed in the feeding artery. Using imaging techniques docetaxel (60 mg/body and 30 mg/body) was infused into the vessels. During chemotherapy the patient received concomitant radiotherapy (50 Gy). MRI after chemoradiation showed a complete response for the primary tumor and a partial response for the neck metastasis. Grade 4 leukopenia and neutropenia and grade 3 pharyngitis/esophagitis were observed during chemoradiotherapy, but these adverse effects abated immediately and were not critical. We conclude that this superselective intra-arterial infusion of docetaxel will be useful and safe for head and neck cancer. PMID- 11865645 TI - [Tumor markers in uterine cancers]. AB - In diagnosing uterine cancers, cells and tissue samples can be directly obtained from the lesion. Cytologic and histologic investigation is the best method for screening and early detection of primary uterine cancers. Tumor markers may be useful for monitoring the clinical course of therapy and early detection of recurrence for which cytologic examination can not be done. Moreover, high levels of tumor markers may represent tumor invasiveness and metastasis to lymph nodes and/or other organs, and may indicate a poor prognosis for the patient. Strictly speaking, tumor markers are not tumor-specific but tumor-associated substances. They can be elevated in sera from healthy individuals under various conditions, and from patients with benign tumors. Squamous cell carcinoma-associated antigen (SCC) is relatively tumor-specific, and widely used for monitoring patients with squamous cell carcinoma not only of the uterine cervix. On the other hand, there is no specific tumor marker for uterine corpus carcinoma. Combination assay of several tumor markers including cancer antigen 125 (CA125) as a core marker may be of greater diagnostic value in cases of uterine corpus carcinoma. PMID- 11865646 TI - Nickel-catalyzed cross-coupling of bromophenols with Grignard reagents in the solid phase synthesis. AB - Polymer-bound substituted bromophenols were found to readily undergo a Ni(0) catalyzed cross-coupling reaction with Grignard reagents to give a variety of substituted phenols and hydroquinones, after cleavage from the support, in moderate to high yields. The requisite bromophenol derivatives, which were prepared from the corresponding phenols and hydroquinone using BTMA Br3 or TBA Br3, were attached to the solid support by the Mitsunobu reaction. PMID- 11865648 TI - Combinatorial chemistry and high-throughput screening in drug discovery: different strategies and formats. AB - Different strategies for the discovery of novel leads interacting with therapeutically relevant targets are thoroughly presented and discussed, using also three recent examples. Emphasis is given to approaches which do not require extensive resources and budgets, but rather prove how cleverness and creativity can provide active compounds in drug discovery. PMID- 11865647 TI - Combinatorial approaches to inhibitors of VLA-4: piperazine-peptoid-bisarylureas. AB - A combinatorial approach towards identifying inhibitors of VALA-4 was investigated. A library of piperazine-peptoid-bisarylureas was assembled in solid phase and screening in the novel v-well assay enabled the identification of active compounds. PMID- 11865649 TI - Combinatorial synthesis of unsymmetrical secondary amines. Application to arylethanolamines, arylpropanolamines and aryloxypropanolamines. AB - Combinatorial libraries of unsymmetrical secondary amines: arylethanolamine, arylpropanolamines and aryloxypropanolamines [1], in particular have been synthesized by four different routes using styrenes, aldehydes, hydroxyaromatic acids and bromoaliphatic acids. The structurally diverse libraries were generated in parallel format using solid phase methodology. The compounds corresponding to prototype I-IV were obtained in high yields and purities. PMID- 11865650 TI - [A retained gauze as a cause of unexplained abdominal complaints]. AB - Three patients, two men aged 36 and 62 years and one woman aged 41 years, developed serious symptoms after abdominal surgery which appeared to be based on a retained gauze. In one patient the interval was 15 years, in the second patient the surgery was a minor procedure and in the third patient a surgical towel was retained in the abdominal cavity. After repeated surgery and complementary treatment the patients made a good recovery. Despite all of the precautions taken during surgical procedures, retained foreign bodies continue to be a clinical problem. This can only be overcome if all the members in an operation team work together meticulously. PMID- 11865651 TI - [The retained surgical sponge, an ongoing surgical problem]. AB - The retained surgical sponge seems to be an ongoing problem. Despite the fact that reports of retained surgical sponges are comparatively rare, this problem seems to be more prevalent than is generally appreciated. Even though counting sponges is a tedious task, it should nonetheless be performed with the utmost attention. Although the presence of radiopaque wires in the sponges is helpful in locating these, it does not prevent surgical sponges from being forgotten. These retained sponges can result in serious conditions such as septic complications or pseudo-tumour formation, which in turn might lead to extensive diagnostic and secondary surgical procedures. PMID- 11865652 TI - [Congenital adrenal hyperplasia: clinical aspects and neonatal screening]. AB - Congenital adrenal hyperplasia (CAH) is a disorder of adrenal steroid synthesis. In 95% of CAH cases, it is caused by 21-hydroxylase deficiency, leading to cortisol deficiency and (in most cases) aldosterone deficiency. The compensatory increase in ACTH secretion by the pituitary gland leads to stimulation of the adrenal glands and, consequently, overproduction of androgens. The classic form is well known due to the congenital virilisation seen in affected girls. However, the cortisol and aldosterone deficiency is at least equally important in both sexes as it can cause an Addisonian crisis within the first weeks of life. For these reasons, a neonatal CAH screening program has been introduced in the Netherlands. Screening results in earlier detection and treatment. The prevalence of the classic form of the disease is 1:12.000 in the Netherlands. Non-classic 21 hydroxylase deficiency is more frequent, presenting with signs of androgen excess from childhood through to adulthood. Treatment of CAH consists of hormonal replacement and surgical correction in case of congenital virilisation in girls. Long-term treatment results, including height at adulthood, have improved over the last decades. Nevertheless, fertility problems can occur in both sexes. PMID- 11865654 TI - [Roaming through methodology. XXXVIII. Common misconceptions involving standard deviation and standard error]. AB - Standard deviation and standard error have a clear mutual relationship, but at the same time they differ strongly in the type of information they supply. This can lead to confusion and misunderstandings. Standard deviation describes the variability in a sample of measures of a variable, for instance the variability in ages of the members of a group. It represents the degree to which the values are scattered around their mean: the higher the standard deviation the wider the spread. The value of the standard deviation is not influenced by the number of observations in the sample. The standard error is always used for extrapolation: to estimate the intervals between which the true value of a statistic will occur, based on a sample of observations and with a certain degree of certainty. When interpreting a standard error, it is important to know which statistic (mean, percentage, relative risk, odds ratio) is to be estimated. The value of the standard error is strongly influenced by the number of observations in the sample: the bigger the sample, the smaller the standard error and the more accurate the estimation. To avoid confusion it is recommended to report no longer the standard error of the mean but instead the confidence intervals of the mean to estimate the true value of the mean. PMID- 11865653 TI - [Gastrointestinal surgery and gastroenterology. XIV. Mesenteric abnormalities in generalised vascular disease]. AB - Various forms of vasculitis may result in mesenteric ischaemia, ischaemic colitis or aneurysm formation in the aorta or intestinal blood vessels. Vasculitides may involve large- and/or medium-sized vessels, medium- and/or small-sized vessels, or small-sized vessels only. It is essential to differentiate between the different forms of vasculitis since diagnostic tests and therapies differ greatly. Gastrointestinal manifestations of vasculitis can generally be detected using angiography, digital subtraction angiography and/or magnetic resonance angiography (MRA). Various laboratory tests are helpful in establishing the diagnosis in patients in whom vasculitis is clinically suspected. In addition, the diagnosis should be confirmed using histology or angiography if possible. Treatment of vasculitis not caused by chronic infection consists of high dose corticosteroids and, in the case of polyarteritis nodosa or vasculitis associated with anti-neutrophil cytoplasmic antibodies (ANCA), cyclophosphamide. PMID- 11865655 TI - [From gene to disease: deafness and connexin 26]. AB - Deafness is genetically heterogeneous, yet it is estimated that approximately half of the heritable cases of autosomal recessive deafness are caused by mutations in the gene coding for connexin 26. Connexin 26 is thought to have an essential role in the transport of potassium ions back to the endolymph of the inner ear after sound stimulation. PMID- 11865656 TI - [Diagnostic image (75). A veterinary assistant with a swelling on the neck. Cowpox]. AB - In a 21-year-old woman, suffering from a moderate painful erythematous infiltrated lesion in the neck, cowpox was diagnosed. Contact with an infected cat was suggested. Healing was spontaneous. PMID- 11865658 TI - [Successful treatment and pregnancy in a women with the non-classic form of congenital adrenal hyperplasia]. AB - In an 18-year-old woman non-classic 21-hydroxylase deficiency was diagnosed and dexamethasone treatment was instituted. Ten years later, she became pregnant for the first time; at 37 weeks unexpected intrauterine foetal death was found to have occurred. A second pregnancy ended with a spontaneous abortion following a 12-week period of amenorrhoea. At the third pregnancy, the medication was replaced with hydrocortisone as it was suspected that the use of dexamethasone may have played a role in the intrauterine foetal death and the spontaneous abortion. The patient gave birth to a healthy, but dysmature, daughter. Female patients with non-classic congenital adrenal hyperplasia present with signs of androgen excess. Treatment with glucocorticoids reduces the symptoms and restores the menstrual cycle and fertility. Preconceptional advice by a clinical geneticist is recommended, because of the risk of an affected child. If there is no risk of having a child with congenital adrenal hyperplasia, hydrocortisone or prednisone is the treatment of choice during pregnancy as neither cross the placenta. PMID- 11865657 TI - [Births and abortions among Amsterdam teenagers according to ethnicity, 1996 1998]. AB - OBJECTIVE: To determine the number of teenage births and abortions in Amsterdam, the Netherlands, in total and according to age and ethnic origin. DESIGN: Retrospective analysis of routinely collected data. METHOD: Based on data collected from the municipal population register and the abortion clinics in Amsterdam, birth rates, abortion rates and abortion ratios were calculated for the period 1996-1998. The birth rate was the number of live births and the abortion rate the number of abortions per 1000 teenage girls. The abortion ratio was the number of abortions per 100 pregnancies. RESULTS: The average birth rate and abortion rate for the period 1996-1998 were 3.2 and 7.6 for 14- to 16-year old girls respectively, and 25.0 and 29.5 for 17- to 19-year-old girls respectively. Among 14- to 16-year-old girls 70 pregnancies out of 100 ended in an abortion and among 17- to 19-year-old girls 54 pregnancies out of 100. In both age groups the birth rate for Surinamese, Antillian, Moroccan, Turkish and Ghanian girls was higher than for Dutch girls. There was one exception: Moroccan girls aged 14-16 years did not have a higher birth rate than Dutch girls of the same age. Surinamese, Antillian and Ghanian girls aged 14-16 years and 17-19 years also had a higher abortion rate compared with Dutch girls of the same age. The Turkish and Moroccan girls did not have (much) higher abortion rates. In both age groups the abortion ratio was lowest for Turkish teenagers and highest for Ghanian teenagers. The figures for second-generation immigrant teenagers were more akin to those of the native Dutch girls than those of first-generation immigrants. CONCLUSION: In Amsterdam unwanted pregnancies were most frequent in Surinamese, Antillian and Ghanian teenage girls. To prevent these pregnancies further research into the (cultural) determinants of sexual behaviour is necessary. PMID- 11865659 TI - [Fainting caused by a neck tumor]. AB - An 86-year-old woman was admitted with progressive complaints of dizziness and the sensation of losing consciousness several times a day; this had led her to fall down on a few occasions. The patient had experienced these symptoms for a month prior to admission. Two weeks before admission her family had noticed a swelling on the left side of her neck. During admission, bradycardia and subsequently asystole occurred while she was eating or when turning her head to the left, followed by a slow junctional escape rhythm. A CT scan of the head and neck region revealed a large tumour in the left parapharyngeal space entrapping the left carotid artery. Histology disclosed a low-grade malignant non-Hodgkin lymphoma, sensitive to radiotherapy. The patient received a pacemaker and treatment of the lymphoma was uneventful. The symptomatic sinus bradycardia and asystole were caused by intermittent carotid massage by the lymphoma. PMID- 11865660 TI - [Roaming through methodology. XXXVI. About 'likelihood' ratios and the rules of Bayes]. PMID- 11865661 TI - [Roaming through methodology. XXXVI. About 'likelihood' ratios and the rules of Bayes]. PMID- 11865662 TI - [Medical mistakes: unavoidable but preventable]. PMID- 11865663 TI - [Erythromycin in premature rupture of membranes beneficial to the health of the child]. PMID- 11865664 TI - Toxicophores: groups and metabolic routes associated with increased safety risk. AB - Drugs and chemicals can be metabolized in living organisms in an effort to remove them from the body. During this excretion process, certain chemicals can be transformed into unstable, highly reactive species. This can result in the overwhelming of cellular defense, allowing the reactive metabolite to bind to cellular macromolecules, and leading to abnormal cellular function. It is widely accepted that this process can lead to clinically observed toxicities in the form of idiosyncratic adverse drug reactions. However, it is currently impossible to predict which chemical species will cause these reactions. This review focuses on drugs from which reactive metabolites have been detected and reiterates the need for a more thorough understanding of basic drug metabolism before attempting to relate chemical species formation to biological function. PMID- 11865665 TI - Towards a drug concentration effect relationship for QT prolongation and torsades de pointes. AB - Preclinical strategies to evaluate torsades de pointes (TdP) have progressed significantly over recent years with reliable and robust in vitro and in vivo methodologies available to assess QT prolongation. Increased emphasis is now being placed on collecting adequate pharmacokinetic data in preclinical studies in order to carry out pharmacokinetic/pharmacodynamic analysis. Free plasma concentrations are being utilized for inter-species comparisons and for relating in vivo and in vitro results, and this approach appears to be optimal. Data obtained in these assays are predictive of the potential of a compound to prolong QT and, by inference, cause TdP. Concentration/effect relationships are apparent such that compounds with positive results in preclinical assays may prove safe, provided that the maximum concentrations expected in the patient population are significantly lower than those required to prolong QT. PMID- 11865666 TI - Models for profiling the potential QT prolongation risk of drugs. AB - The appearance of QT prolongation and arrhythmic events associated with a compound undergoing clinical trials can greatly hamper drug development programs. Assessing the risk of a compound during preclinical studies to cause this cardiotoxicity is thus critically important to the pharmaceutical industry. A wide variety of preclinical approaches exist to evaluate potential QT issues, including in vitro, in vivo and in silico (i.e., computer simulation) methods. We present an evaluation of recent reports implementing these techniques, with an emphasis on the linkage between drug-induced cardiac action potential changes and QT prolongation both in vitro and in silico. We conclude with a strategy that integrates in silico modeling with in vitro and in vivo experimentation to create a compelling package for assessing potential proarrhythmic risk of a compound. PMID- 11865667 TI - Improving structure-linked access to publicly available chemical toxicity information. AB - Publicly available toxicity databases serve as the central resource in efforts to develop algorithms for assessing potential chemical toxicity. File standardization and linkage of chemical structures with chemical toxicity information are essential first steps in providing broad access to existing toxicity information, for deriving useful structure-activity relationship (SAR) models, performing analog searches, and estimating the potential toxicity of new chemicals. This review will focus on current efforts to improve structure-linked access to publicly available sources of toxicity information, outlining current web-based resources as well as two new database initiatives for standardizing and consolidating public chemical toxicity information. PMID- 11865668 TI - Genotype/phenotype comparisons: a probe for the effect of disease progression on drug metabolism. AB - The effect of disease state on drug metabolism has been investigated using the relationship between genotype and metabolic phenotype. The two polymorphic probes, N-acetyltransferases-2 (NAT2) and cytochrome P450 2C19 (CYP2C19), were respectively used in HIV+/AIDS patients and patients with advanced cancer. The results of the studies suggest that advanced disease produces discordances between genotype and phenotype, indicating a reduction in the metabolic capabilities of these individuals. Thus, polymorphic enzymes such as CYP2C19 and NAT2 can be used to probe changes in drug-metabolizing enzyme capacities. The development of genotype/phenotype discordances should reflect general changes in metabolic capabilities and, thus, alterations in the activities of other important enzymes such as CYP3A. The data also suggest that the genotype/phenotype probes can be used to optimize the clinical treatment of patients with advanced disease states. PMID- 11865669 TI - Structural insights into the promiscuity and function of the human pregnane X receptor. AB - The pregnane X receptor (PXR) is a promiscuous nuclear receptor that responds to a wide variety of drugs, xenobiotics and endogenous compounds, and plays a critical role in mediating drug-drug interactions in humans. PXR is the master regulator of the expression of the CYP3A4 gene, which encodes for the most abundant and promiscuous drug-metabolizing enzyme in humans. PXR also regulates the expression of other genes involved in xenobiotic metabolism, including CYP2C8, CYP2C9, CYP2B6, GSTA2 and MDR1, as well as genes critical to bile acid metabolism. While PXR functions as a xenobiotic sensor in numerous vertebrates, its relatively low sequence conservation across species causes the PXRs from different organisms to respond to distinct subsets of xenobiotics. Thus, PXR promiscuity is directed and not random. The recent determination of crystal structures of the ligand binding domain of human PXR has provided the first detailed molecular view of this promiscuous receptor, and has advanced our understanding of its varied biological functions. We review the evidence establishing the binding promiscuity of PXR and its directed specificity in different species, and analyze the structural determinants of these characteristics. In addition, we examine the relationship between the interaction of PXR with ligands and the manner in which CYP3A4 is thought to bind to substrate molecules. The accumulating structural and functional data on PXR may facilitate the development of improved methods for in vitro, in vivo and in silico screening for PXR activation. PMID- 11865670 TI - Web alert. The chemistry of metabolic and toxicological processes. PMID- 11865671 TI - High-throughput and in silico techniques in drug metabolism and pharmacokinetics. AB - The high-throughput screening (HTS) of large proprietary compound collections and combinatorial libraries has increased the pressure on gathering pharmacokinetic and drug metabolism data as early as possible. Properties related to absorption, distribution, metabolism and excretion (ADME) can be estimated by a range of in vivo and in vitro methods, most of which are now available or under development in high(er)-throughput modus. In addition, progress has been made in in silico methods using various quantitaTive structure-activity relationship (QSAR) and molecular modeling techniques that employ a range of recently introduced descriptors tailored to e-ADME. These in silico approaches are promising filters for virtual libraries to aid synthesis as well as the selection of compounds for acquisition and screening in the early stages of drug discovery. PMID- 11865672 TI - Evolutionary computational methods to predict oral bioavailability QSPRs. AB - This review discusses evolutionary and adaptive methods for predicting oral bioavailability (OB) from chemical structure. Genetic Programming (GP), a specific form of evolutionary computing, is compared with some other advanced computational methods for OB prediction. The results show that classifying drugs into 'high' and 'low' OB classes on the basis of their structure alone is solvable, and initial models are already producing output that would be useful for pharmaceutical research. The results also suggest that quantitative prediction of OB will be tractable. Critical aspects of the solution will involve the use of techniques that can: (i) handle problems with a very large number of variables (high dimensionality); (ii) cope with 'noisy' data; and (iii) implement binary choices to sub-classify molecules with behavior that are qualitatively different. Detailed quantitative predictions will emerge from more refined models that are hybrids derived from mechanistic models of the biology of oral absorption and the power of advanced computing techniques to predict the behavior of the components of those models in silico. PMID- 11865673 TI - Automated sample preparation and LC-MS for high-throughput ADME quantification. AB - Bioanalytical groups in the pharmaceutical industry provide quantitative data to support all stages of drug discovery. The increased use of 96-well plates and robotic liquid handling systems, the availability of robust triple quadruple mass spectrometers, and developments in chromatographic and samples preparation techniques, have all increased the rate at which this data can be generated. This review describes currently used methods and emerging technologies for automation of high-throughput quantitative bioanalysis. The focus is on recent applications of sample preparation and chromatography techniques compatible with detection by triple quadruple mass spectrometers. PMID- 11865674 TI - Intravenous administration of poorly soluble new drug entities in early drug discovery: the potential impact of formulation on pharmacokinetic parameters. AB - Currently, in early drug discovery, compounds that are formulated for first animal experiments are increasingly characterized as being lipophilic and poorly water-soluble. Typical examples of intravenous formulations for these compounds include aqueous solutions at non-physiologically high or low pH, co-solvent solutions, solutions in cyclodextrins (CDs), surfactant-based solutions, mixed micellar solutions, parenteral fat emulsions or nano- and microsuspensions. Experiments designed to determine the intrinsic pharmacokinetic behavior of a new drug entity (NDE) are complicated as, depending upon the formulation, disposition in the organism can be affected. This may be due to slow or incomplete dissolution of injected particles, precipitation in the bloodstream, delayed release from the dosing vehicle, competition between compound and formulation ingredients for transport and metabolism mechanisms, or altered binding to blood components. The most important determinant for the successful development of a 'non-interfering' dosing vehicle is the required dose. Provided the analytical technique used to determine drug concentration in the body is sensitive enough to allow compound administration at low doses, screening formulations at comparatively low concentrations may be feasible. In this way, formulation approaches that may potentially impact the pharmacokinetic behavior of the compound of interest can be avoided. PMID- 11865675 TI - Increasing throughput in lead optimization in vivo toxicity screens. AB - Lead optimization requires toxicity screening strategies to select a compound with a high likelihood of successful development. As numerous compounds need to be screened and resources to direct toward any single compound are limited, short turnaround times to generate and interpret data are needed. Utilization of in vivo toxicity screens is necessary for an effective screening strategy, however, if not appropriately implemented, they may consume excessive resources and prolong selection of a developable compound. Optimization of in vivo studies requires identifying effective placement into the screening strategy, selecting the appropriate study designs, implementing processes that allow rapid data generation and interpretation, and understanding the utilities of in vivo data. When implemented, an effective, high-throughput screening strategy will utilize adequate but minimal amounts of resources, and will prioritize processing near technical time limits. These require generating only the data from which decisions will be made and can be best achieved using as few animals as possible per study. PMID- 11865676 TI - Gene expression profiling for pharmaceutical toxicology screening. AB - Advances in medicinal chemistry and high-throughput pharmacological screening are creating a multitude of potential lead compounds. There is also heightened concern about drug-induced toxicity, which is all too often uncovered late in development or at the post marketing stage. Together, these factors have created a need for novel approaches to screen for toxicity. There have been technological advances that enable study of changes in the gene expression profile caused by toxic insults and important steps made toward unraveling target organ toxicity at the molecular level. Thus, gene expression profile-based screens hold the promise to revolutionize the way in which compounds are selected for development. For screens focused on specific mechanisms of toxicity, reporter gene systems have proven utility, albeit modest because of our limited knowledge of which genes are true surrogate markers for toxicity. For broader forecasts of toxicity, DNA microarrays hold great promise for delivering practical gene expression profile screens (GEPS). For this promise to be realized, however, a number of technological hurdles must be cleared: (i) cost; (ii) reproducibility; (iii) throughput; and (iv) data analysis. Of equal if not greater importance, issues relating to the test systems used, the requisite number of genes to be studied and the size and scope of the database upon which forecasts will be based must be addressed. At present, the proof-of-concept for GEPS for toxicity is in hand, and we are poised to realize the goal of creating practical GEPS for application in compound prioritization. PMID- 11865677 TI - Transcriptomics in predictive toxicology. AB - Once again, genomics is about to change drug development. Following its major impact on target discovery and assay development, which increased the number of compounds at early stages of the process, genomics is now zeroing in on the prediction of potential toxicological problems of compounds. Toxicogenomics is the analysis of toxicological processes at the transcriptome level of a target organ or cell. By simultaneously monitoring the effect of a compound on the transcription levels of hundreds to thousands of genes, toxicogenomics can provide an enormous amount of data. This data bears information on the way in which compounds act at the molecular level, reaching far beyond the mere conclusion of whether or not a particular toxicological outcome is elicited. By compiling transcription profiles for well-known toxicants, we are beginning to learn how to analyze this novel type of data in the context of mechanistic and predictive toxicology. PMID- 11865678 TI - Analysis of gene and protein expression for drug mode of toxicity. AB - The rapid sequencing of the genomes of a number of organisms, including humans, has led to major changes in the drug industry. The abundance of genome data, and the reagents generated from these genomes, have enabled the study of changes in large numbers of genes and proteins in parallel, using methods such as DNA microarrays to examine gene expression changes, or 2D polyacrylamide electrophoresis (2D-PAGE) to observe changes in the expression of proteins. While these techniques have been in use for several years, their application has primarily focused on the target discovery phase, with some early work carried out on drug- or toxin-induced changes in proteins using 2D-PAGE. In the last two years, a slew of publications have appeared on the application of array technologies to the study of toxicology, and the aim of this review is to highlight some recent examples of these applications. PMID- 11865679 TI - Longitudinal left ventricular function in a population of healthy adults: a tissue Doppler imaging study. AB - BACKGROUND: Longitudinal left ventricular function is a major determinant of global ventricular function and is probably more sensitive than radial function in the detection of disease. Tissue Doppler imaging of the mitral annulus allows the study of longitudinal left ventricular function. METHODS: Forty-five healthy volunteers divided into two groups (younger and older than 45 years) were studied with pulsed tissue Doppler imaging of the 4 sides of the mitral annulus (septal, lateral, inferior, anterior) in 4 and 2 apical chamber views. In each wave (systolic-s, rapid filling-e, atrial contraction-a) we analyzed velocities, time intervals and velocity-time integrals, as well as heterogeneity and asynchrony indexes. Data were compared between the different sides in each group, between groups and with conventional Doppler data. RESULTS: In contrast to the septal side, the lateral side of the annulus shows higher velocities and velocity-time integrals of the s and e waves, with non-significantly shorter isovolumic relaxation time and shorter time to peak e. There is functional agreement between the lateral and inferior sides versus the septal and anterior sides of the annulus. Most systolic parameters remained unchanged with aging; however, aging was associated with decreased e velocity, increased a velocity and inverted e/a ratio. The relation between s and fractional shortening also did not change with aging. Annular isovolumic contraction time and isovolumic relaxation time were shorter than their respective global time intervals. CONCLUSIONS: This study shows that there are physiological differences in velocities, time intervals and velocity-time integrals between the 4 sides of the mitral annulus, reflecting physiological heterogeneity and asynchrony, and that some of these parameters are age dependent. These data also contribute to a better understanding of longitudinal left ventricular function and may be useful in future studies as reference values in control groups. PMID- 11865680 TI - Study of myocardial function: why tissue Doppler. PMID- 11865681 TI - Cardiovascular manifestations in Marfan syndrome. AB - INTRODUCTION: Cardiovascular complications are the major cause of morbidity and mortality in Marfan syndrome (MS), a common connective tissue disorder. Currently it is considered that the prognosis and morphologic characteristics in infantile Marfan syndrome may be quite different from those reported in older patients. The objective of this study was to analyze the cardiovascular manifestations and evolution of the patients with Marfan syndrome followed at our pediatric cardiac unit. METHODS: The authors reviewed the clinical files of all the patients that fulfilled the diagnostic criteria for MS according to De Paepe et al. (1996). We analyzed the following parameters: gender, age at referral, race, family history, clinical examination, diagnostic exams, therapy and evolution. The patients were divided into two groups according to the age at diagnosis: infants (group 1) and older patients (group 2). RESULTS: Group 1 included 3 infants, two boys and one girl, sporadic cases, presenting congestive heart failure. The major cardiac diagnoses were aortic dilatation (1/3) and mitral valve prolapse with severe mitral regurgitation (2/3). Congenital heart disease was associated in two cases (patent ductus arteriosus and atrial septal defect). Two needed cardiac surgery at an early age and one was recently proposed for surgery. There were no deaths. Group 2 included 20 patients, 14 boys and 6 girls, first seen at a mean age of 8 years. Ten had a positive family history and none presented cardiac symptoms. The major cardiac manifestations were mitral valve prolapse (18/20) and aortic dilatation (17/20). There was no significant progression of the cardiac lesions, except for one case, during the 12 years of follow-up. CONCLUSION: Infantile MS presented high morbidity; mitral regurgitation was severe in two cases. All patients presented heart failure, two needing early operations. In classic MS evolution was favorable, and the commonest cardiac lesions were mitral valve prolapse and aortic dilatation. We emphasize the need for beta-blockers to prevent progression of aortic dilatation. The decision for surgery rests upon the severity of valve regurgitation and the rate of progression of aortic dilatation. It is important to inform patients and family about physical exercise, prevention of endocarditis, risks associated with pregnancy and genetic counseling. PMID- 11865682 TI - Prognosis in patients with heart failure and preserved left ventricular systolic function. AB - BACKGROUND: It is recognized that heart failure patients with preserved left ventricular systolic function have better prognosis; nevertheless, there are some studies with conflicting results. Also, there is a paucity of data concerning the prognostic factors in this group of patients. OBJECTIVES: To determine possible variables with prognostic relevance in heart failure patients with preserved left ventricular systolic function (ejection fraction > 40%). METHODS: 157 consecutive ambulatory patients with heart failure were assessed; those patients with ejection fraction > 40% were included in the study (n = 46). All patients were evaluated by clinical interview and physical examination, ECG, echocardiogram (M mode, 2D and pulsed Doppler of mitral flow), biochemical study and determination of type B natriuretic peptide (BNP). The patients were grouped according to the rhythm presented on ECG: Group I--patients with atrial fibrillation; Group II- patients in sinus rhythm Group II was further subdivided in two groups according to the presence or absence of restrictive left ventricular filling pattern. All patients had a clinical follow-up, with recording of events (death or hospitalization from cardiac cause). The mean follow-up time was 682.2 +/- 55 days. RESULTS: The mean age of the patients was 70.4 +/- 1.2 years; 54.3% were women; mean ejection fraction was 49.6 +/- 1%; mean BNP levels were 202.9 +/- 41.3 pg/ml. Mortality was 19.6% and the combined event death or hospitalization from cardiac cause) occurred in 26.1% of the patients. Among the clinical, demographic, biochemical, echocardiographic and neurohumoral parameters, only BNP levels had prognostic significance in the whole population. In Group II patients, BNP levels, heart rate and restrictive left ventricular filling pattern were identified as having prognostic significance. Kaplan-Meyer curve analysis showed that both BNP and restrictive left ventricular filling pattern seemed to be important prognostic markers. CONCLUSIONS: This preliminary study suggests thar neurohumoral activity (determined by plasma BNP levels) and a restrictive ventricular filling pattern may be important factors in prognostic stratification of heart failure patients with preserved left ventricular systolic function. PMID- 11865683 TI - Biventricular pacing--early experience. AB - INTRODUCTION: Biventricular pacing has been studied for the treatment of chronic heart failure (CHF). This technique seems to be able to improve symptoms and exercise tolerance, in patients with advanced CHF and prolonged QRS duration. OBJECTIVE: To present our experience with biventricular pacing in the management of severe CHF. METHODS: Between June 2000 and March 2001, 8 patients with dilated cardiomyopathy and left bundle branch block (LBBB) were selected for transvenous biventricular pacing system implantation. Mean age: 54.12 +/- 16.8 years; 5 males. The etiology was: idiopathic in 6 cases; operated congenital heart disease in 1, and ischemic in the other. Despite tailored treatment of CHF (with all patients taking diuretics and angiotensin-converting enzyme inhibitors), 7 patients remained in NYHA class III and 1 in IV. The quality of life score (assessed by the "Minnesota living with heart failure questionnaire") was 62.25 +/- 11.29. Seven patients had sinus rhythm and only one chronic atrial fibrillation; mean PQ duration--220 +/- 76.37 ms; mean QRS duration--168.75 +/- 20.31 ms. RESULTS: Implant failure, due to coronary sinus dissection and to excessive fluoroscopy time, with no coronary sinus catheterization, occurred in 2 cases (success rate: 75%). Implant data: mean implant procedure duration: 122.5 +/- 47.82 min; mean fluoroscopy time: 35.66 +/- 22.06 min; QRS duration, after implant: 133.33 +/- 15.05 ms. Left ventricular lead final position: anterolateral in 2 patients and lateral in 4. Pacing thresholds: biventricular--1.36 +/- 0.6 V; right ventricle--0.28 +/- 0.04 V; right atrium--0.32 +/- 0.08 V. Pacing impedance (left ventricle): 1013.33 +/- 147.87 omega. Follow-up (1st and 3rd month): one patient died, suddenly, 15 days after the procedure. In the others, an improvement in the quality of life index and functional class was found. These results were independent of echocardiography data. There were no significant differences in the pacing threshold and impedance during the follow-up. CONCLUSIONS: Patients with advanced CHF and widened QRS benefited from biventricular stimulation, in which improvement on the clinical status was evident. PMID- 11865684 TI - Echocardiographic patterns and prognosis in heart failure. AB - BACKGROUND: There are many variables with prognostic value in patients with heart failure (HF). Those related to left ventricular function are among the most important. Recently, the evaluation of the patterns of ventricular filling by pulsed Doppler echocardiography has been studied as a variable with prognostic value. OBJECTIVES: To evaluate the prognostic value of echocardiography variables (diastolic and systolic) in patients with HF. These variables were analysed in respect to hospital admission for cardiovascular reasons or death. MATERIAL AND METHODS: We evaluated 157 consecutive patients with HF and included 110 patients who were in sinus rhythm. The mean age was 68.2 +/- 0.9 years. HF was ischemic in 52.7%. Patients underwent echocardiography examination within the week of reference. The patients were grouped according to left ventricular (LV) systolic dysfunction (LV ejection fraction < 40%). We also classified patients in two groups according to the presence of a restrictive pattern in diastolic transmitral flow profile. Finally, we classified all patients in four groups according to their systolic function and diastolic pattern: Group I--systolic dysfunction and restrictive ventricular filling pattern. Group II--systolic dysfunction without restrictive ventricular filling pattern. Group III--without systolic dysfunction with restrictive ventricular filling pattern. Group IV- without systolic dysfunction without restrictive ventricular filling pattern. The events were death or hospital admission. The mean follow up time was 625 +/- 55 days. We did a statistical analysis and for all tests a p value < 0.05 was considered statistically significant. RESULTS: We found impaired LV systolic function (systolic HF) in 73.6% and restrictive ventricular filling pattern in 45.5%. During the follow-up 41.8% died or were admitted to hospital. Patients with systolic HF had lower admission free survival rate. Patients with restrictive ventricular filling pattern had lower admission free survival rate than those without. Group I had lower admission free survival rate than Group II and Group IV. Group IV had a higher admission free survival than all other Groups. CONCLUSIONS: These results support and expand previous observations that diastolic function variables, such as the pattern of ventricular filling (namely the restrictive) have independent prognostic value in patients with HF. PMID- 11865686 TI - Heart failure treatment in Portuguese hospitals: results of a survey. AB - The management of heart failure in Europe is largely conducted by primary care physicians in out-patient clinics and by cardiologists and internists in hospitals. Several reports suggest differences among these specialists regarding knowledge and actual practice, and indicate that the application of guidelines is far from optimal. In order to look for differences between cardiologists and internists in terms of implementation of guidelines a survey was carried out among the directors of 83 hospital departments of cardiology and internal medicine in Portugal. The survey included questions about diagnostic and treatment protocols, special areas for management, and suggestions to improve the quality of heart failure patient treatment. The answers suggest that in Portuguese hospitals at least half of the patients with HF are treated by internists. Treatment protocols exist in about 25% of the cardiology departments but are virtually non-existent in internal medicine. The use and availability of echocardiography are high in cardiology but no more than reasonable in internal medicine. There are neither special in-hospital areas nor specialized nurses for the treatment of HF. Cardiologists recognize the need for greater specialization in this field--doctors, nurses and clinics--but this is not a priority for internists. An effort should be made to improve in-hospital HF treatment. PMID- 11865685 TI - Poisoning with calcium channel blockers--a case report and review of the literature. AB - The incidence of poisoning with calcium channel blockers, accidental or intentional, has increased in recent years, associated with more frequent use. We present a clinical case of bradycardia and shock of unknown cause, which came to be revealed a poisoning by 3240 mg of slow-release diltiazem, managed with temporary transvenous pacing and dopamine in high concentration. We make a review of the cardiovascular manifestations of the three classic calcium channel blockers: verapamil, diltiazem and nifedipine; namely, hypotension, rhythm and conduction disturbances. We point out the late appearance of the beginning of manifestations with the use of slow releasing formulations. The toxicity by calcium channel blockers can lead to a wide variety of manifestations in the central nervous system, gastrointestinal system, endocrine-metabolic, hematologic and respiratory systems. There is a high clinical suspicion when the following factors are present: hypotension with bradycardia, mental state disturbances, lactic acidosis, hyperglycemia, sinus pauses and refractory shock. Treatment is based on general measures of intoxication support, decreasing the drug absorption and improvement of cardiac function. The bradyarrhythmias are corrected with the use of intravenous calcium, glucagon, atropine and pacemaker. If the intoxication causes depression of cardiac contractility, the use of calcium or/and glucagon is indicated. If there is refractoriness with these measures, catecholamines should be employed. There are alternative and adjuvant drugs such as amrinone, insulin glucose, 4-aminopyridine and calcium entry promoters. Charcoal hemoperfusion can be useful in the overdose of sustained release preparations, but hemodialysis is unworthy of therapeutical interest. PMID- 11865687 TI - Basic principles on selection and use of diagnostic tests: properties of diagnostic tests. AB - In clinical practice, in order to design and implement a specific therapeutic plan, as well as communicating an appropriate prognosis, the doctor needs to establish a precise diagnosis of the condition. Sometimes all one needs is a clinical impression. More often, however, the definition of an accurate diagnosis will mandate the interpretation of specific diagnostic tests as well. The rational use of diagnostic tests in cardiology--whether laboratorial or imaging- should be based on three factors: 1) validity of results of studies on the test; 2) diagnostic properties of the test; and 3) applicability of the test in the clinical setting. The rational use and the correct interpretation of diagnostic tests are based on these three factors. In a previous article we presented the basic principles concerning the validity of the results from the study that defined the specific test, and what level of evidence that constitutes. In this article we present the diagnostic properties of tests (sensitivity, specificity, positive and negative predictive values, likelihood ratios, odds). Finally, in a forthcoming paper we will discuss the applicability of the test in clinical cardiological practice. PMID- 11865688 TI - Unusual image. PMID- 11865689 TI - [Long-term use of a left ventricular assist device for end-stage heart failure]. PMID- 11865690 TI - [Essence and significance of surgery. Part I -- Birth of surgery: determining factors and contexts]. AB - The author intends to particularly analyse the origin of Surgery as regards its deterministic factors and contexts, resounding the essence and the meaning of Surgery itself. The primary core of the surgical practice dates back to Prehistoric Times, when, driven by his self-preservation instinct, the cave man, when suffering from some trauma, performed on himself a series of more or less immediate "actions" in order to remain healthy. At the same time, a second meaningful nucleus of the surgical experience rises contiguously to the operations the Prehistoric Man performed on another member of his clan. The third stage of this ongoing process, coincident with the origin of surgery in the strict sense of the word, goes back to the tribal context: in fact, in this social organisation only one member of the group was specifically assigned to treat diseases, based on group regulations. For the mediterranean area, the chronological development of this evolution is likely to have started 250,000 years ago in connection with the experience initially of Neanderthal Man and subsequently Cro-Magnon Man in Pleistocene and Holocene of the Quarternary Era respectively, and it could have finished at the beginning of the Neolithic, when the "ancient civilization" of the Mediterranean Basin arose in approximately 10,000 B.C. PMID- 11865691 TI - [Clinically non-palpable lesions of the breast: radiologic features, biologic factors, and surgical strategy]. AB - The use of mammography for early detection of breast cancer showed an increased detection of non-palpable breast-lesions (NPBL). The authors evaluate the radiologic findings, the biological factors and the surgical approach, trough the personal experience and the literature, for a correct treatment of these lesions. PMID- 11865692 TI - [Chromoendoscopy in the diagnosis of precancerous lesions of the esophagus. Our experience]. AB - The elements of an unfavourable prognosis for oesophageal cancer are frequent metastasis, high incidence of local recurrence and mainly the difficulty of an early diagnosis. Alcohol, tobacco and precancerous lesions are the most important risk factors of these tumours. According to literature, the authors suggest the method of chromoendoscopy, with vital staining by lugol or blue toluidine for endoscopic; guidance to biopsy in the aimed screening of patients, whose habits- alcohol, smoking--should cause, in time, the rising of lesions with neoplastic potentiality. Endoscopy with bioptic test is the best diagnostic investigation. In fact sensibility and specificity of these investigations increase using this method with vital staining. PMID- 11865693 TI - [Malignancy: treatment and prognosis of gastrointestinal and pancreatic endocrine neoplasia: retrospective study of a series of 16 cases]. AB - The aim of this study was to review our experience with endocrine tumours of the gastrointestinal tract and pancreas (ETGIP). Between February 1991 and March 2000, sixteen patients with ETGIP were operated on at our institution. Of these patients we reviewed preoperative symptoms, diagnostic techniques (ultrasound, CT, MRI, radiolabelled octreotide scintigraphy, angiography, immunohistochemical study), treatment (surgical operation, neoadjuvant and adjuvant chemotherapy, and radiometabolic therapy) and survival. Nine patients (56%) had a carcinoid tumour, three (19%) an unspecified endocrine tumour, and four (25%) an endocrine tumour associated with a non-endocrine neoplasm. Only five patients (31%) had a preoperative diagnosis of endocrine tumour. Eight patients (50%) had metastatic disease at the time of the operation. All patients without preoperative metastasis (eight patients, 50%) are still alive without recurrent disease, with a mean postoperative survival of 36 months (12-60 months). Of eight patients with metastatic disease, six (75%) died after a mean of 20.5 months (3-60 months) and two (25%) are still alive with the disease after 3 and 6 months, respectively. These data show that presence of metastasis strongly influence survival. Furthermore, survival of patients with metastatic disease seems to be longer as compared to other gastrointestinal tract malignancies. ETGIP are more common and aggressive than previously believed and, therefore, early diagnosis is crucial for cure. Nowadays, however, new diagnostic tools such as radiolabelled octreotide scintigraphy are available for diagnosis and postoperative follow-up. The optimal treatment for ETGIP is a multimodal approach with surgical operation, chemoradiation, radiometabolic, and genetic therapies. PMID- 11865694 TI - [Study of the possibility of an effect of biliary disease on the occurrence of chronic pancreatitis]. AB - The authors report a series of patients suffering from chronic pancreatitis and underline the high frequency of biliary pathology in these subjects. The authors remind, then a research performed, some years ago, by one of them. In such a research the performing of an anastomosis between the pancreatic duct and the biliary common duct in a series of experimental animals caused an acute pancreatitis or in the surviving animals a chronic pancreatitis. The authors believe that a biliary pathology may be responsible not only of an acute pancreatitis but also of a chronic pancreatitis. PMID- 11865695 TI - [Indications and surgical treatment of acute complicated diverticulitis]. AB - Although the surgical treatment of acute and complicated diverticulitis has been refined over the years, discussion is still ongoing about whether a single or two stage procedure (Hartmann's procedure) should be performed in patients with peritonitis from perforation of a colonic diverticulum. A retrospective review was completed for patients undergoing surgery for acute complicated diverticulitis from 1980 to 1997. All patients were categorized according to Hinchey classification. Of the 186 patients treated, emergency operation was performed in 52 (28 per cent; group I) and an elective operation in 134 (72 per cent; group II). All patients had a resection of the involved colon and anastomosis was performed using CEEA staplers. In the group I, 50 patients underwent resection with immediate reconstruction with or without colostomy and two an Hartmann's procedure. Among the group II, primary anastomosis with or without proximal diversion was performed in 132 patients; two patients underwent an Hartmann's procedure. Post-operative mortality was of 5.76% in the group I and 2.9% in the group II. Nine anastomotic leaks occurred in the group I and 18 in the group II. Comparing these results with the recent literature, we have concluded that primary resection is virtually always possible in acute and complicated diverticulitis. Primary anastomosis with or without colostomy, in expert hands, is a safe procedure for patients in stage I of Hinchey's classification or II, but should be considered on an individual basis in presence of peritonitis. In case of fecal peritonitis, persistent hypotension, ascites, severe immune compromise and extreme malnutrition the Hartmann procedure represent the procedure of choice. PMID- 11865696 TI - [Surgical treatment of acute colonic diverticulitis. Personal experience]. AB - BACKGROUND: Colonic acute diverticulitis presents in patients that suffered from colonic diverticula in 10-25% of cases higher percentage in the elder patient. Colonic acute diverticulitis mortality leads high rates in over 70 years old patients with a value a upper than 75-80%. Perforation and diffuse peritonitis are the more serious complications. Accurate clinical staging followed by correct surgical option are the base of the therapeutical success. METHODS: Between january 1995 and june 2000 we studied by a retrospective analysis 56 patients (34M/22F), mean age 67, hospitalized cause acute perforate colonic diverticulitis, all patients were recorded in order their clinical conditions APS score and Hinchey staging of the disease. At the time of the recovery all patients underwent at clinical examination, abdominal ultrasound, plane X-ray, and bloody tests. Surgical operations performed were: PRA; PRA and protective colostomy; Hartmann procedure. In the patients with APS score > 6-10 and aged > 70 Hartmann procedure was performed. RESULTS: Overall mortality was 9% (5 pazienti), overall morbility was 40%. On the basis of the I and II Hinchey stage mortality rate was 0%. In the III stage was recorded mortality of 4% and 33% in IV stage. Complication were: 6 anastomotic lekage; 7 infections of the laparotomic wound; 1 incisional hernia; 2 lekeages of the stomoa; 3 stomatitis; 5 proctitis. CONCLUSION: Therapeutical choise treatment has to consider Hinchey stage and APS score. Hartmann procedure is indicated in stage III-IV with score APS > 8-10. For the other patients primary resection anastomosis and if is necessary associated colostomy is the treatment recommended. PMID- 11865697 TI - [Functional results of colorectal and coloanal anastomosis with and without pouch]. AB - In the last two decades one of the main targets of anorectocolonic surgery has been to develop sphincter saving procedure able to achieve good results with acceptable five-years survivals, optimal local control of the diseases and low rate of local cancer recurrence. Partially the development of new operative techniques such as low colorectal and coloanal anastomoses with or without pouch, the TME operation and the nerve sparing procedure have reach this target. In fact, often after these operations we can observe a functional syndrome called "Post Anterior Resection Syndrome". The basis of this syndrome have to researched in anatomical and physiological alterations that followed a reconstructive operation. It is characterized by frequency and fragmentation of the stool, feeling of incomplete evacuation, tenesmus and urgency. Fecal continence may be compromised to different levels: usually with alteration limited to soiling and impaired control of flatus, occasionally with loss of liquid stool, rarely with loss of solid stools. The anorectal function will be altered for long time following the surgical procedure and the stabilization of functional results may require 1-3 years. On the basis of these considerations, the authors examine the etiopathogenesis and clinical presentation of the "Post Anterior Resection Syndrome", suggesting some expedients to prevent the functional problems. Analysing our experience and a wide specific bibliography, they also underline the indispensable point to achieve a good functional results after a reconstructive procedure. The author conclude asserting that the absence of these points have to be carefully valued because, in these situations, a simply colostomy is able to guarantee a better quality of life that a colorectal/coloanal anastomoses with or without pouch but associated to functional problems. PMID- 11865698 TI - [Polypropylene mesh in the surgical treatment of inguinal and crural hernia: our experience with 500 cases]. AB - The authors guided by their personal experience (612 abdominal hernioplasties performed between 1992 and 1998) after using for the cure of inguinal and femoral hernias diverse prosthesis illustrate the characteristics of the polipropene prosthesis which in their opinion possesses every requirement necessary to enable it to become the defining ideal prosthesis. The authors underline again the use of this routinely applied prosthesis, also in a local anaesthesia it has favoured the development of the tension-free and suture-less technique which hes allowed hernioplasty to spread in day-surgery. PMID- 11865699 TI - [Tension-free lumbar hernioplasty]. AB - The authors report a case of a 51 year-old woman with spontaneous Grynfeltt's hernia. Computerized tomography accurately displayed the anatomy of the lumbar region, thus aiding diagnosis and treatment. A double layer polypropylene mesh was used for the repair, placed in the pre peritoneum and sutured to the margins of the hernial defect avoiding muscles and fascias. The use of a double layer prosthesis and reduction of tension on the suture line allow a sound repair, with minimal postoperative pain and immediate resumption of daily activities. PMID- 11865700 TI - [Use of PTF prosthesis in hernia in the elderly]. AB - The paper reports the author's experience of the use intraperitoneal of Gore-Tex Dual Mesh Biomaterial in large incisional hernia operations in patients over seventy. From January 1999 to December 1999 we operated on 24 patients for treatment of abdominal wall defect. In all patients we used a Dual Mesh Plus Biomaterial. Overall mortality was 0. Morbility was 9.5%. Follow-up is too short for definitive considerations about the incidence of recurrences but our initial experience with this material encourages us to use it again for replacement of abdominal wall defects in the age. PMID- 11865701 TI - [Robotic and systems technology for advanced endoscopic procedures]. AB - The advent of endoscopic techniques changed surgery in many regards. This paper intends to describe an overview about technologies to facilitate endoscopic surgery. The systems described have been developed for the use in general surgery, but an easy application also in other fields of endoscopic surgery seems realistic. The introduction of system technology and robotic technology enables today to design a highly ergonomic solo-surgery platform. This consists of a system of devices for endoscopic surgery (HF, light source, etc...) with which the surgeon interacts directly, positioning systems for optic and instruments that the surgeon drives as the likes without assistance, and a chair to increase the comfort of the surgeon during surgery. The system of endoscopic devices named OREST (Dornier, Munchen) designed already in 1992 opened the way to a number of systems available today that allow to the surgeon a direct control of the instrumentation. A considerable step ahead in endoscopic technology is the introduction of robotic technology to design assisting systems for solo-surgery and microsurgical instrument manipulators. Results of a number of experimental trials on combinations of different positioning devices are presented and commented. A further step in the employment of robotic technology is the design of "master-slave manipulators" to provide the surgeon with additional degrees of freedom of instrumentation. In 1996 a first prototype of an endoscopic manipulator system, named ARTEMIS, designed in cooperation with the Research Center in Karlsruhe, could be used in experimental applications. Clinical use of the system, however, will require further development of the arm mechanics and the control system. The combination with the implementation of telecommunication technology will open new frontiers, such as teleconsulting, teleassistance and telemanipulation. PMID- 11865702 TI - [Multiple choledocholithiasis]. AB - OBJECTIVE: The authors report their experience about the endoscopic (ERCP) treatment of multiple choledocholithiasis. DESIGN: Report of 5 cases; evaluation of mortality and effectiveness of the treatment. SETTING: Operative Unit of General and Thoracic Surgery, Department of Surgical, Anatomical and Oncological Disciplines. Policlinico, University of Palermo. INTERVENTIONS: ERCP + ES + clearance of bile duct stones in all 5 patients. RESULTS: 100% successful. CONCLUSION: ERCP is choice treatment of multiple choledocholithiasis, only in well experienced teams. PMID- 11865703 TI - [Inflammatory markers after laparoscopy versus laparotomy cholecystectomy]. AB - OBJECTIVE: The study is carried out to determine whether the level of IL-6 is altered and in what way after surgery as well as if such a change could be an indicator of increased morbidity after surgical treatment. MATERIALS OF THE STUDY: Chemiluminesence immunoeassay system was used in order to establish IL-6 level in blood samples of 71 patients that underwent abdominal surgery, 36 Laparoscopic Cholecystectomy (LC) and 35 Open Cholecystectomy (OC) at time 0 (before the operation), 1 h, 2 h, 3 h, 6 h, 24 h and 48 h after the operation. RESULTS: Plasma IL-6 levels are significantly increased after OC; we observed 3 cases of post-operative infections, in which IL-6 returned to normal levels 6 days after surgery. Analogous variation to the IL-6 levels was noted for the C reactive protein levels. DISCUSSION: Laparoscopic cholecystectomy, a so called mini-invasive surgical procedure, is associated to a small increase of IL-6 serum levels and provides better post-operative conditions to the patients by reducing surgical stress and the infectious complications correlated to the surgical procedure. CONCLUSIONS: During OC there is a significant higher elevation of IL-6 serum levels than after laparoscopic cholecystectomy. Variation of C-reactive protein serum levels after surgery is analogous to variation of IL-6 levels. PMID- 11865704 TI - Th1-like and Th2-like cytokines in patients undergoing open versus laparascopic cholecystectomy. AB - The advantages of laparoscopic (LC versus, open cholecystectomy (OC) seems to be related to minimal invasive procedure and to the moderate inflammatory response. The aim of this study is to define the involvement of Th1 (IFN-gamma) and Th2 (IL 4, IL-6, IL-10, IL-13) cytokines production in vivo and in vitro in patients undergoing OC or LC. In 42 patients undergoing LC (n = 22) and OC (n = 20) Th1 like and Th2-like was evaluated before operation and at 6, 24 and 48 hours after operation for white blood cell counting and cytokines (IL-4, IL-6, IL-10, IL-13, IFN-gamma, TNF-alpha) in the sera and in the supernatants from circulating mononuclear cells stimulated with phytohemagglutinin or lipopolysaccharide. The acute phase response cytokine, IL-6, appeared significantly increased following OC than after LC. All other cytokines did not very significantly. In vitro data shows a reduction of IFN-gamma and increase in Th2-like cytokines in OC patients compared with the basal value. In LC subjects we observed an high production of IFN-gamma associated to an increase of Th2-like cytokines, like IL-10 and IL-13, even though IL-4 and IL-6 were unmodified. In contrast to OC, LC did not significantly affect immunocompetence, maintaining a moderate inflammatory response and an adequate balance between Th1 and Th2 cytokine. Furthermore, the strong activation of cells producing Th1-like cytokines in LC patients following mitogen activation indicated a consistent anti-microbial activity, that was not detectable in OC patients, that showed after activation only a Th2 response. PMID- 11865706 TI - When the plumbing failed: an allegory. PMID- 11865705 TI - [Chordoma. Diagnostic and therapeutic problems]. AB - The chordoma is a rare mesodermic tumor derived from the notochord which arises and growths inside the vertebral bodies. It is a slow development tumor with late clinical manifestations, rarely metastatic often with local reoccurrences. The treatment of choice of this tumor is surgical and the access must be individualized to the single clinical case: anterior, posterior or combined antero-posterior respect to the spine. Recently videolaparoscopy has been proposed when an anterior approach is indicated without increased morbidity or mortality. Chemotherapy is not indicated because low tumoral responsness. Radiotherapy is indicated as a palliative procedure when a surgical approach ca't be radical. Its application is useful to treat pains and to control the post operative course increasing the disease-free interval. The authors report the case of an old symptomatic lady with a chordoma in the sacral region. The patient underwent subtotal absportation an anterior transperitoneal approach. Because the extension of the tumor, its biological characteristics and the patient's age the authors adopted this less invasive approach. PMID- 11865708 TI - Interactions between vitamins and drugs. PMID- 11865707 TI - Brain function in fetal alcohol syndrome assessed by single photon emission computed tomography. AB - This case series reports results of single photon emission computed tomography (SPECT) studies in three patients with fetal alcohol syndrome (FAS), who had previously undergone structural magnetic resonance imaging (MR). The MR studies revealed several brain anomalies, including microcephaly, agenesis or hypoplasia of corpus callosum and agenesis of hippocampal commissure. The SPECT data revealed that the CBF was reduced by at least 25% in the temporal region relative to the cerebellum in all three patients. By contrast, the temporal-cerebellar differences were between 4% and 7% in two controls. The functional abnormalities in FAS, like neuroanatomical abnormalities, are likely to be multiple and varied because of heterogeneity of this syndrome. Our findings suggest the need for a larger study to test the hypothesis that temporal lobe abnormalities are a notable occurrence in FAS. Discovery of specific regional brain dysfunctions (such as temporal lobe dysfunction) that are particularly vulnerable to alcohol's teratogenic effect may allow clinicians and researchers to look for markers useful in FAS screening and may have implications for prevention and treatment of FAS. PMID- 11865709 TI - Genealogical inference of closely related species based on microsatellites. AB - Despite their unmatched popularity in many research areas, microsatellites have not yet become a major tool for the inference of genealogical relationships of closely related species. Recent studies have successfully extended the repertoire of microsatellite analysis beyond population genetics and demonstrate that phylogenetic relationships of closely related species can be inferred accurately with fewer loci than previously assumed. PMID- 11865710 TI - Map-based cloning of quantitative trait loci: progress and prospects. AB - Map-based cloning has been considered problematic for isolating quantitative trait loci (QTLs) due to the confounding phenotypic effects of environment and other QTLs. However, five recent studies, all in plants, have succeeded in cloning QTLs using map-based methods. We review the important features of these studies and evaluate the prospects for broader application of the techniques. Successful map-based cloning requires that QTLs represent single genes that can be isolated in near-isogenic lines, and that genotypes can be unambiguously inferred by progeny testing. In plants or animals for which map-based cloning of genes with discrete phenotypes is feasible, the modified procedures required for QTLs should not be limiting in most cases. The choice between map-based cloning and alternative methods will depend on details of the species and traits being studied. PMID- 11865711 TI - Numb suppresses the negative complementation at the Notch locus of Drosophila melanogaster, suggesting a putative mechanism for negative complementation. AB - The mutant form of the intracellular asymmetrically localized Numb membrane-bound protein of Drosophila melanogaster suppresses the negative complementation of certain Abruptex (Ax) mutations of the Notch (N) locus encoding a transmembrane receptor protein in which the Ax mutations are mutations in the epidermal growth factor (EGF)-like repeats of the extracellular domain of the receptor. One model for how Ax mutants affect N function is that they are refractory to an antagonistic signal generated by an excess of N ligands. Genetically numb (nb) is an antagonist of N. In the absence of nb, cells follow the same fate as they would in the presence of a gain-of-function N allele, such as Ax. Numb has been shown to interact with the cytoplasmic domain of Notch. It is therefore suggested that numb counteracts the effect of Abruptex on Notch ligand binding, i.e. that Numb is an antagonist to the activation of the Notch signal generated by Notch ligands. Numb might accomplish this by interfering with the proteolytic cleavage of the Notch intracellular domain at the cell membrane. Thus, it seems possible that the mechanism of negative complementation of certain Ax mutants is the failure of this cleavage. Other possible mechanisms for negative complementation are also discussed. PMID- 11865712 TI - The genetic basis of larval resistance to a host plant toxin in Drosophila sechellia. AB - The larvae of Drosophila sechellia are highly resistant to octanoic acid, a toxin found in D. sechellia's host plant, Morinda citrifolia. In contrast, close relatives of D. sechellia, D. simulans and D. melanogaster, are not resistant. In a series of interspecific backcrosses, 11 genetic markers were used to map factors affecting egg-to-adult ('larval') resistance in D. sechellia. The third chromosome harbours at least one partially dominant resistance factor. The second chromosome carries at least two mostly dominant resistance factors but no recessive factors. However, neither the X chromosome--which contains 20% of D. sechellia's genome--nor the fourth chromosome appear to affect resistance. These data suggest that larval resistance to Morinda toxin may involve only a handful of genes. These results, when compared with a previous analysis of adult resistance to Morinda toxin in D. sechellia, suggest that larval resistance may involve a subset of the genes underlying adult resistance. PMID- 11865713 TI - Control of female pheromones in Drosophila melanogaster by homeotic genes. AB - We have investigated the role of the Antennapedia and Bithorax complexes (ANT-C and BX-C) on the production of cuticular hydrocarbons in Drosophila melanogaster. In males, there is little, if any, influence of these complexes on the hydrocarbon pattern. In females, there are large and opposite effects of these complexes on diene production: two ANT-C mutations cause an increase in diene production and a reduction of monoenes, whereas most BX-C mutations result in a decrease in dienes and an increase in monoenes, although their sum remains constant. The effect is the highest in Mcp and iab6 females. It is suggested that a factor originating from the prothorax might activate the conversion of monoenes to dienes in females. The abdomen seems to have a crucial role in the production or control of pheromones: abdominal segments four to seven have the main effects, with a most dramatic effect for segments four and five. PMID- 11865714 TI - A pseudo-likelihood method for estimating effective population size from temporally spaced samples. AB - A pseudo maximum likelihood method is proposed to estimate effective population size (Ne) using temporal changes in allele frequencies at multi-allelic loci. The computation is simplified dramatically by (1) approximating the multi-dimensional joint probabilities of all the data by the product of marginal probabilities (hence the name pseudo-likelihood), (2) exploiting the special properties of transition matrix and (3) using a hidden Markov chain algorithm. Simulations show that the pseudo-likelihood method has a similar performance but needs much less computing time and storage compared with the full likelihood method in the case of 3 alleles per locus. Due to computational developments, I was able to assess the performance of the pseudo-likelihood method against the F-statistic method over a wide range of parameters by extensive simulations. It is shown that the pseudo-likelihood method gives more accurate and precise estimates of Ne than the F-statistic method, and the performance difference is mainly due to the presence of rare alleles in the samples. The pseudo-likelihood method is also flexible and can use three or more temporal samples simultaneously to estimate satisfactorily the NeS of each period, or the growth parameters of the population. The accuracy and precision of both methods depend on the ratio of the product of sample size and the number of generations involved to Ne, and the number of independent alleles used. In an application of the pseudo-likelihood method to a large data set of an olive fly population, more precise estimates of Ne are obtained than those from the F-statistic method. PMID- 11865715 TI - Population bottleneck and effective size in Bonamia ostreae-resistant populations of Ostrea edulis as inferred by microsatellite markers. AB - Genetic variability at five microsatellite loci was analysed in three hatchery propagated populations of the flat oyster, Ostrea edulis. These populations were part of a selection programme for resistance to the protozoan parasite Bonamia ostreae and were produced by mass spawns, without control of the genealogy. Evidence for population bottlenecks and inbreeding was sought. A reduction in the number of alleles, mainly due to the loss of rare alleles, was observed in all selected populations, relative to the natural population from which they were derived. Heterozygote excesses were observed in two populations, and were attributed to substructuring of the population into a small number of families. Pedigree reconstruction showed that these two populations were produced by at most two spawning events involving a limited number of parents. Most individuals within these populations are half or full-sib, as shown by relatedness coefficients. The occurrence of population bottlenecks was supported by estimates of effective number of breeders derived by three methods: temporal variance in allelic frequencies, heterozygote excess, and a new method based on reduction in the number of alleles. The estimates from the different methods were consistent. The evidence for bottleneck and small effective number of breeders are expected to lead to increasing inbreeding, and have important consequences for the future management of the three O. edulis selected populations. PMID- 11865716 TI - Two-generation analysis of pollen flow across a landscape. III. Impact of adult population structure. AB - The rate and distance of instantaneous pollen flow in a population are parameters of considerable current interest for plant population geneticists and conservation biologists. We have recently developed an estimator (phi ft) of differentiation between the inferred pollen clouds that fertilize several females, sampled within a single population. We have shown that there is a simple relation between phi ft and the average pollen dispersal distance (delta) for the case of a population with no geographic structure. Though forest trees usually show considerable pollen flow, assuming an absence of spatially distributed genetic structure is not always wise. Here, we develop analytical theory for the relation between phi ft and delta, for the case where the probability of Identity by Descent (IBD) for two individuals decreases with the physical distance between them. This analytical theory allows us to provide an effective method for estimating pollen dispersal distance in a population with adult genetic structure. Using real examples, we show that estimation errors can be large if genetic structure is not taken into account, so it is wise to evaluate adult genetic structure simultaneously with estimation of phi ft for the pollen clouds. We show that the results are only moderately affected by changes in the decay function, a result of some importance since no completely established theory is available for this function. PMID- 11865717 TI - Estimating genotypes with independently sampled descent graphs. AB - A method for estimating genotypic and identity-by-descent probabilities in complex pedigrees is described. The method consists of an algorithm for drawing independent genotype samples which are consistent with the pedigree and observed genotype. The probability distribution function for samples obtained using the algorithm can be evaluated up to a normalizing constant, and combined with the likelihood to produce a weight for each sample. Importance sampling is then used to estimate genotypic and identity-by-descent probabilities. On small but complex pedigrees, the genotypic probability estimates are demonstrated to be empirically unbiased. On large complex pedigrees, while the algorithm for obtaining genotype samples is feasible, importance sampling may require an infeasible number of samples to estimate genotypic probabilities with accuracy. PMID- 11865719 TI - Statistical power of QTL mapping methods applied to bacteria counts. AB - Most QTL mapping methods assume that phenotypes follow a normal distribution, but many phenotypes of interest are not normally distributed, e.g. bacteria counts (or colony-forming units, CFU). Such data are extremely skewed to the right and can present a high amount of zero values, which are ties from a statistical point of view. Our objective is therefore to assess the efficiency of four QTL mapping methods applied to bacteria counts: (1) least-squares (LS) analysis, (2) maximum likelihood (ML) analysis, (3) non-parametric (NP) mapping and (4) nested ANOVA (AN). A transformation based on quantiles is used to mimic observed distributions of bacteria counts. Single positions (1 marker, 1 QTL) as well as chromosome scans (11 markers, 1 QTL) are simulated. When compared with the analysis of a normally distributed phenotype, the analysis of raw bacteria counts leads to a strong decrease in power for parametric methods, but no decrease is observed for NP. However, when a mathematical transformation (MT) is applied to bacteria counts prior to analysis, parametric methods have the same power as NP. Furthermore, parametric methods, when coupled with MT, outperform NP when bacteria counts have a very high proportion of zeros (70.8%). Our results show that the loss of power is mainly explained by the asymmetry of the phenotypic distribution, for parametric methods, and by the existence of ties, for the non parametric method. Therefore, mapping of QTL for bacterial diseases, as well as for other diseases assessed by a counting process, should focus on the occurrence of ties in phenotypes before choosing the appropriate QTL mapping method. PMID- 11865720 TI - [Serving the public]. AB - What is meant by Public Service? Does it mean serving the public or, since the service is for everyone, must the public leave it up to the discretion of the administration, in the hopes of obtaining what it is entitled to? The question is important at a time when the public is no longer satisfied with a simple service, but demands acknowledgement, quality and performance from the general Administrations and in particular hospital institutions. With an aim for global efficiency, the annual report of the General Authority of Social Affairs "Les Institutions socials face aux usagers", has courageously tackled the eternal items: information for the public, administration of the users, quality of service rendered. This recent report offers the opportunity to review, from the users' point of view, the quality of public services and, with the malaise in this sector today, this represents a major social stake in the short-term. PMID- 11865718 TI - The power of the transmission disequilibrium test (TDT) with both case-parent and control-parent trios. AB - The transmission disequilibrium test (TDT) customarily uses affected children and their parents (often case-parent trios, TDTD). Control-parent trios are necessary to guard against spurious significant results due to segregation distortion but are not generally utilized in the identification of disease susceptibility loci (DSL). Controls are often easy to recruit and the TDT can easily be extended to include control-parent trios into the analyses with unrelated case-parent trios. We present an extension of the TDT (TDTDC) that incorporates unrelated cases and controls and their parents into a single analysis. We develop a simple and accurate analytical method for computing the statistical power of various TDT (e.g. the TDTD, TDTDC, TDTDC and TDTC that employ control-parent trios only) under any genetic model. We investigated the power of these TDT, and particularly compared the relative power of the TDTD and TDTDC. We found that the TDTDC is almost always more powerful than the TDTC and TDTD. The relative power of the TDTDC and TDTD depends largely upon a number of parameters identified in the study. This study provides a basis for efficient use of control-parent trios in DSL identification. PMID- 11865721 TI - [Micro-angiopathic and psychological risk in children and adolescents with type 1 diabetes]. AB - OBJECTIVE: Evaluate the correlation between psychopathological disorders in type 1 insulin-dependent diabetic (IDD) children and adolescents and the occurrence of micro-angiopathic complications. METHODS: Two hundred and five IDD children (104 girls and 101 boys with a mean age of 13.4 +/- 3.5 years on inclusion) were longitudinally monitored between 1994 and 2000. Somatic complications were systematically searched for during a yearly control visit to the day hospital. Psychiatric disorders had been assessed initially by self-administered questionnaires filled-in by the children (Spielberger's STAIC-Trait for anxiety, CESD for depression and Child-EAT for eating habits), by the parents (CBCL, CPRS or GHQ-28) or the principle teachers (CTRS) and diagnostic screening of psychopathological disorders every year. RESULTS: More than 140% of the IDD children (n = 86) were diagnosed DSM-IV, with predominant anxiety (n = 41), during the longitudinal evaluation. Positive screening was significantly associated with higher psychopathological scores before inclusion in the study, with children considered more perturbed by the parents, teachers and the children themselves and mothers who considered themselves more perturbed (GHQ-28 mean 5.6 +/- 7.5 vs. 3.5 +/- 4.8; t = 2.211 p = 0.028). These children were less compliant. Disorders were associated with metabolic imbalance, measured by HbA1c, overweight and an excess of micro-angiopathic, particularly retinopathic, complications. CONCLUSION: Our results show the frequency of mental disorders in IDD children, the importance of the parents' psychopathology and its association with increased somatic risk. The nature of this relationship requires clarification and the incidence of prevention measures targeted on the psychological problems. The interest of associating a paedopsychiatric element in the therapeutic management of these patients and their families is emphasized. PMID- 11865722 TI - [Granulomatous arteritis through sarcoidosis]. AB - INTRODUCTION: Digestive localisation of sarcoidosis is rare. OBSERVATION: A 35 year-old man presented with sarcoidosis revealed by a mediastinal hilum lymphadenopathy 13 years earlier. Epigastric pain led to oeso-gastroduodenal fibroscopy and biopsies, showing inflammatory mucosa and numerous giant-cell epithelioid granulomas, without concomitant necrosis or fibrosis. COMMENTS: The clinical manifestations and endoscopic profile of gastric localisations of sarcoidosis are not specific. Diagnosis relies on several elements: presence of epithelioid granulomas without necrosis, history of sarcoidosis or the simultaneous existence of other localisations, evocative biological signs and the absence of elements evoking any other diagnosis. Treatment relies on corticosteroid therapy and sometimes requires endoscopic or surgical management. PMID- 11865723 TI - [Role of endoprostheses in transluminal coronary angioplasty. Practice evaluation during the first half of 1997 and 1998 at the Mulhouse Hospital Center]. PMID- 11865724 TI - [Terminal renal failure in France. The situation of chronic renal insufficiency in France]. PMID- 11865725 TI - [Chronic renal insufficiency: towards a public health policy]. PMID- 11865726 TI - [Epidemiology of care demands]. AB - SOME FIGURES: Terminal renal failure (TRF) is a major public health problem in France in view of its increasing incidence (110 pmi/year), prevalence (700 pmi) and the costs of treatments supplied. In France, more than 6,500 new patients started treatment for TRF in 2001; around 42,000 patients with renal failure have been treated. The mean cost of treatment per patient is estimated to be of 350 KF per annum for dialysis, 450 KF for transplantation the first year and 50 KF per annum thereafter. Hence, more than 10 billion francs are spent every year on treating TRF, i.e., 1.5% of the Health Scheme. However, these costs do not include expensive treatments (erythropoietin), transport or hospitalisation. RECENT TENDENCIES: Our information system concerning TRF is fragmented and not coordinated. Identification of the cases is incomplete, their declaration is not always systematic and the quality control of the data has not been formalized. Nonetheless, major tendencies can be identified. The notable facts of the last 10 years are an aging TRF population and an increase in associated comorbidity. The diseases leading to TRF are changing. Vascular nephropathies predominate; ischemic renal diseases have become the first cause of TRF in elderly patients. The incidence of type 2 diabetes is increasing and strangely in the French overseas territories. Glomerular nephropathies are the third cause of TRF, particularly in the young. However, compared with other causes, their prevalence is decreasing. INSUFFICIENCIES: The morbidity and mortality with dialysis is dominated by cardiac and vascular causes. Renal transplantation has stagnated; the waiting lists increase and donations are insufficient. Conversely, transplantation survival is progressing. In a second part, we will examine the elements of health strategy necessary to adapt the supply of care and the organization of preventive measures. PMID- 11865727 TI - [Supply of care and prevention]. AB - THE SITUATION: The needs for care of terminal renal failure (TRF) has changed over the past 10 years, particularly with more elderly patients presenting with more comorbid states. The supply of care, the quota of which is fixed by the demand, is no longer adapted and reorganization is primordial. The delayed referral to a nephrologist is an increased source of morbidity and mortality. It is also the cause of increased costs of around 0.2 billion francs per year. In our paper we present 2 major axes of development, defined during the last National Health Conference. ORGANIZATION OF A NETWORK ON NEPHROLOGIC EPIDEMIOLOGY AND INFORMATION (RESEAU EPIDEMIOLOGIE ET INFORMATION EN NEPHROLOGIE -REIN): This network will pilot the development of the first axe, which is the supply of care adapted to requirements. The characteristics of this multidisciplinary program are presented together with the multi-source information system. The provisional annual budget for dialysis alone would correspond to the contribution of 1.00 FF per session for the Health Scheme and to 0.6 FF for the State. SECOND AXE OF DEVELOPMENT: Transplant planning is aimed to increase the collection of organs and thus the yearly number of transplants, clearly insufficient today. The principle elements of the program are presented. The yearly return on investment of such a program would be of half a billion Francs after five years. PREVENTION OF TRF: The notion of renal risk, risk factors for the progression of chronic renal failure and risk factors for a nephropathy must be introduced. There are two major axes to the prevention of TRF: slowing the progression of chronic renal failure and preventing the occurrence of subjacent nephropathies. These tasks come within the framework of the "Chronic Renal Failure Plan" presented by the Ministry of Health last September. PMID- 11865729 TI - Leg ulcer care: nursing attitudes and knowledge. PMID- 11865728 TI - Family-centred care in the NICU. PMID- 11865730 TI - Caring for survivors of the Holocaust. PMID- 11865731 TI - The forensic ABCs of trauma care. PMID- 11865732 TI - The legal risks of e-mail. PMID- 11865733 TI - Article captured nurses' frustrations. PMID- 11865734 TI - Vacancies to fill. PMID- 11865735 TI - Storming through the stigma. PMID- 11865736 TI - How supportive? PMID- 11865737 TI - Thanks for stigma article. PMID- 11865738 TI - Address stigma with education. PMID- 11865739 TI - Erasing term's gender association. PMID- 11865740 TI - A challenge.... PMID- 11865741 TI - Call me registered nurse. PMID- 11865742 TI - Screening mammography in older women: a pilot study of residents' decisions. AB - BACKGROUND: Screening mammography is a commonly employed preventive modality. The employment of mammography in older women is not supported by evidence from clinical trials, largely because elderly women were excluded from such trials. Most guidelines do not recommend routine screening in older women. How residents reason in this gray zone has been subject to little empirical study. PURPOSES: This study sought to answer two questions: How variable are residents' decision responses to mammography screening scenarios in older women where there is no clear evidence? What reasons do residents give to justify these decisions? METHODS: Residents were asked to respond to four scenarios and give their screening recommendations and the reasons justifying their decisions. RESULTS: There was considerable variability in resident responses to the four scenarios. Only in one scenario was there near unanimity on the preferred screening decision. Resident perceptions of quality of life, longevity and understanding of the guidelines were cited as justification for their decisions. CONCLUSION: Clinical preceptors should be aware of how the variability of resident perceptions of such factors as quality of life and prognosis may influence decision-making. PMID- 11865744 TI - Is that your final answer? Relationship of changed answers to overall performance on a computer-based medical school course examination. AB - BACKGROUND: Whether examinees benefit from the opportunity to change answers to examination questions has been discussed widely. PURPOSE: This study was undertaken to document the impact of answer changing on exam performance on a computer-based course examination in a second-year medical school course. METHODS: This study analyzed data from a 2 hour, 80-item computer delivered multiple-choice exam administered to 190 students (166 second-year medical students and 24 physician's assistant students). RESULTS: There was a small but significant net improvement in overall score when answers were changed: one student's score increased by 7 points, 93 increased by 1 to 4 points, and 38 decreased by 1 to 3 points. On average, lower-performing students benefited slightly less than higher-performing students. Students spent more time on questions for which they changed the answers and were more likely to change items that were more difficult. CONCLUSIONS: Students should not be discouraged from changing answers, especially to difficult questions that require careful consideration, although the net effect is quite small. PMID- 11865743 TI - Communication between consultants and referring physicians: a qualitative study to define learning and assessment objectives in a specialty residency program. AB - BACKGROUND: Outpatient consultation constitutes a major part of medical practice. However, little is known about the skills which should be taught to residents in order for them to improve their consultant-referring physician relationships. PURPOSES: To specify the consultant skills which are required to ensure an effective communication between specialists and referring physicians. METHODS: A qualitative study based on (a) a literature search and (b) focus group interviews. RESULTS: Skills thus identified and described are classified in two groups: observable skills and principles/attitudes. CONCLUSIONS: The consensual specification of these abilities permits a greater efficacy in the teaching of consultant skills. PMID- 11865746 TI - A statistical technique for the development of an alternate list when using constrained optimization to make admission decisions. AB - BACKGROUND: In an earlier study it was demonstrated that constrained optimization could be used to accurately translate admission goals. Constrained optimization differs from weighting equations in that it does not assign a rank ordering. Although constrained optimization is conceptually superior, some procedures within the admissions process require a rank ordering of applicants. PURPOSE: The purpose of this study is to describes and evaluate the use of two discriminant analysis procedures to obtain a rank order list by generating weights that model constrained optimization procedures. This study also evaluates the usefulness of an additional method that does not require a rank ordering. METHODS: Premium Solver selected a class from the 1998/99 applicant pool. A discriminant analysis was used to generate a discriminant function for modeling the dichotomous group classification selection variable. These weights were then applied and the discriminant function values calculated. The success of the procedure was evaluated by examining rank orders and the magnitude of the correlation and R square statistic. RESULTS: Discriminant analysis accounted for 70% of the decision variance generated by the constrained optimization procedure. Using real data allowed an estimate of the number of students impacted by inconsistent outcomes. CONCLUSION: Discriminant analysis could be used to manage an alternate list, however it will be based on somewhat different criteria than the initial selection procedure. Each method evaluated has advantages and disadvantages and the selection of one method over another depends on what outcomes are most valued by the college. PMID- 11865745 TI - Learning the thyroid examination--a multimodality intervention for internal medicine residents. AB - BACKGROUND: Many physicians have inadequate physical diagnosis skills and cannot detect thyroid abnormalities on physical examination. PURPOSE: To evaluate a multimodality intervention to improve thyroid examination skills using a prospective controlled trial in first-year residents enrolled in an academic internal medicine program. METHODS: The intervention group received a 60-minute educational session during which an endocrinologist described anatomical landmarks, thyroid abnormalities, and examination techniques using a slide show, computerized animation, videotape, and live demonstration on a volunteer with goiter. Residents examined a normal and a goitrous thyroid under the observation of a preceptor and received an evidence-based handout on the thyroid examination. The control group received no specific intervention. Examination technique and identification of thyroid abnormalities were blindly assessed in 2 stations of an objective structured clinical examination (OSCE). RESULTS: Of the 19 residents in the intervention group and the 20 in the control group, 6 (32%) and 3 (15%), respectively, observed the neck for thyroid abnormalities (P = 0.3), 17 (90%) and 16 (80%) used proper hand position (P = 0.7), and 13 (68%) and 15 (75%) had the patient swallow while the neck was palpated (P = 0.7). There was a significant difference in the mean scores based on thyroid physical findings during the OSCE between the intervention and control groups (100 vs. 52.5 [maximal possible score = 200], P = 0.047). CONCLUSION: A 1-hour multimodality learning session furthered the ability of first-year internal medicine residents to detect thyroid abnormalities. PMID- 11865747 TI - Validity of admissions measures in predicting performance outcomes: the contribution of cognitive and non-cognitive dimensions. AB - BACKGROUND: Admissions committees face the daunting task of selecting a small number of candidates who are most likely to succeed in medical school from a large pool of seemingly suitable applicants. While numerous studies have shown moderate correlations among measures of academic performance, predictors of the non-cognitive domain (e.g. interpersonal, communication, ethical) remain elusive, in part because of the absence of a sound criterion measure. PURPOSE: We examined the utility of several cognitive and non-cognitive criteria used in the admissions processes in predicting both cognitive and non-cognitive dimensions of the licencing examinations of the Medical Council of Canada (LMCC). METHODS: Predictors included: undergraduate GPA, undergraduate science GPA, an autobiographical letter, scores from a simulated tutorial, a personal interview and the MCAT. Of specific interest was the relation between measures of communication and problem-exploration skills as assessed during the admissions process and Part II of the LMCC Examination, a multi-station OSCE. RESULTS: Undergraduate GPAs were found to have the most utility in predicting both academic and clinical performance. Scores derived from the simulated tutorial did not predict future performance. The MCAT Verbal Reasoning score and the personal interview were found to be useful in predicting communication skills on the LMCC Part II. CONCLUSIONS: The results have implications for any school that uses the interview as an admissions tool. PMID- 11865748 TI - Validity of admissions measures in predicting performance outcomes: a comparison of those who were and were not accepted at McMaster. AB - BACKGROUND: In typical validity studies, regression analyses are used to examine the relation between admissions measures and subsequent performance. This approach is problematic as it generally yields low correlation coefficients, which are difficult to interpret. Further, it leaves unanswered the question of how those applicants rejected by the process would fare had they been admitted. PURPOSE: This study examines the validity of the admissions measures used to assess non-cognitive qualities at McMaster's Medical School in a unique manner. METHODS: Three cohorts: (a) those offered an admission on the first round, (b) those offered an admission on the second round and (c) those rejected by McMaster, but accepted to another Canadian medical school were compared on admissions evaluations and licencing examination performance. RESULTS: The results indicate that although the scores of those who were offered an admission were significantly greater than those rejected by McMaster on each of the admission tools, licencing examination performance was comparable. CONCLUSIONS: These results are consistent with a previous regression-based validity study and indicate the need for closer examination of admissions tools. PMID- 11865749 TI - Constructivism: the view of knowledge that ended philosophy or a theory of learning and instruction? AB - BACKGROUND: Constructivism is referred to in two very different ways in education including medical education: to refer to a view of knowledge and to refer to a theory of learning and hence instruction. SUMMARY AND CONCLUSIONS: This proposal (a) distinguishes between these two usages of constructivism and (b) concludes that constructivism is not a theory of learning and thus as such has little to offer that might be of value to medical education. PMID- 11865750 TI - The feasibility and acceptability of implementing formal evaluation sessions and using descriptive vocabulary to assess student performance on a clinical clerkship. AB - BACKGROUND: A systematized approach to descriptive evaluation of clinical performance using a vocabulary of global descriptors in the setting of formal evaluation and feedback sessions has been shown to be reliable and valid. The feasibility of this method beyond the institution at which it was developed has not been studied. PURPOSE: To determine the feasibility and acceptability of implementing formal evaluation and feedback sessions, using a vocabulary of global descriptors, in a third-year core clinical clerkship. METHODS: In 1997, the University of Utah internal medicine clerkship introduced an evaluation method in which student performance was discussed at formal sessions, using a taxonomy of global terms describing progressive development from "reporter" to "interpreter" to "manager/educator" (R-I-M-E). The sessions were face-to-face meetings between the clinical teachers and a clerkship director, at three-week intervals through the twelve-week clerkship at the inpatient teaching sites. Following the evaluation session students met individually with the clerkship director for feedback. To determine feasibility, the authors estimated the time and resources necessary to administer the system and recorded actual teacher attendance over 2 academic years (1997-99). Anonymous surveys, using a four point, Likert-type scale, were used to determine acceptance of the method by faculty, residents, and students. RESULTS: Attendance was high for residents (79%) and faculty (72%). Mean survey responses from residents and faculty rated the descriptive system "more valid" than the previous method. A majority of the students rated the method as either "helpful" (30%) or "very helpful" (50%). Time requirement for eight to ten students at each teaching site, for evaluation and feedback sessions was one-half day per week of the clerkship director, every three weeks. CONCLUSION: Our experience establishes the feasibility of implementing this system of formal evaluation and feedback, using descriptive vocabulary, beyond the institution at which it was developed. Students, residents, and faculty endorsed this evaluation system and the survey results suggest substantial utility and face validity. PMID- 11865751 TI - Theories of knowledge and instructional design: a response to Colliver. PMID- 11865752 TI - The White Coat Ceremony: turning trust into entitlement. AB - PURPOSE: To examine the symbolism of the white coat and the nature of authority as employed in the White Coat Ceremony for incoming first-year medical students. Little has been written concerning the rapid spread, bioethical implications or theoretical underpinnings of this ceremony despite adoption by over 100 medical schools in the United States, international recognition by medical schools in Israel and as the subject of recent editorial opinion in the United Kingdom. SUMMARY: The short white coat can be a highly useful tool allowing patients to identify practitioners in a liminal state. However, by officially sanctioning the white coat as a sign of the psychological contract of professionalism and empathy, the medical establishment may be responding to abrogations of its own authority and is teaching students that they are respected for their sartorial behavior separate from their behavior as individuals. CONCLUSION: The White Coat Ceremony fosters a sense of entitlement whereby authority based on title and uniform, and authority based on trust, are poorly distinguished. PMID- 11865753 TI - [New trends and attitude in vascular medicine]. AB - The author discusses in four points (indications in etalons, reduction of invasiveness, extension of indications, new technologies) the tremendous scientific progress that has been achieved nowadays in the field of cardiovascular surgery. A special emphasis is given to the role of interventional cardio-angiology that can be practiced only in surgical departments with suitable profile. Illustrative cases are presented along with a list of the most pertinent references. It is finally suggested that the mastering of the invasive and tissue sparing techniques is also a task of cardiovascular surgeons. A co-operation is indispensable. In the field of this new branch of science further developments are necessary in Hungary. The employment of the most up to date technology can be recommended and accepted only after the collection of one's own initial experiences. The introduction of the new technologies requires a substantial financial expenditure that will be hopefully recovered in terms of the expected considerable reduction of cardiovascular mortality which is at present 55% in Hungary. PMID- 11865755 TI - [Transposition of ovaries during radical hysterectomy for cervical cancer]. AB - INTRODUCTION: 422 radical hysterectomies were performed for cervical cancer between 1st July 1990. and 31st December 2000. in the 1st Department of Obstetrics and Gynecology, Semmelweis Medical University. At least one of the ovaries was conservated and suspended in 21 cases. AIM(S): The aim of this study is to preserve ovaries guaranteeing natural oestrogen production for young patients suffering in early stage cervical cancer, and to prove appropriate efficacy of their method. PATIENTS/METHOD(S): According to present examinations transposition of ovary (21 patients) and radiotherapy made as a part of the protocol can only slightly influence the endocrine function of ovary. RESULTS: Only in one case of 21 a hormonal substitution was needed for treating menopausal syndrome. Studying histopathologically ovaries removed during 422 radical hysterectomies the authors could not find any metastasis given by early stage (I/A2 or I/B) cervical cancer. Following and controlling their patients after ovarial transpositions for a long period they could not observe any pathological ovarial disorder (cyst). CONCLUSION(S): According to the data of international literature the authors conclude, that transposition of ovaries during radical hysterectomy for well-selected, younger aged, early staged cervical cancer cases can result only a low risk, but much better quality of life for the patients. PMID- 11865754 TI - [Acute gastrointestinal bleeding in patients with antithrombotic therapy]. AB - Approximately one-fifth of the patients, referred to the acute gastrointestinal bleeding unit took antithrombotic drugs (anticoagulants and antiplatelet agents) in a dose or with an effect, which made the causal relationship with acute bleeding episodes unequivocal. This paper analyzes the data of the first 100 such patients of the ward. The majority used acethyl-salicylic acid derivatives, however, a substantial number were on coumarol, and some bleeding patients with prosthetic heart valves were also observed. Haemostasis was forced by urgent, accurate, intervention based endoscopy along with simultaneous rapid efforts to correct the underlying clotting abnormalities. Prosthetic valve patients needed special care and attendance, and the reinstitution of anticoagulant treatment as soon as possible to maintain valve patency. The experiences and schedules of the authors, as well as the proposals of the literature are reviewed and summarized. PMID- 11865756 TI - [Acute abdomen in primary care]. AB - The author--using his 30 years practical medical experience, from the point of view of a family doctor giving the first diagnose, analyses the problem of acute abdomen. Gives an overview of anamnesis (heteroanamnesis) and the data and their significance investigated during the physical examination of patient (inspection, palpation, percussion, auscultation), and that of the usefulness for the right diagnose. The author orients the reader's attention to the fact, that the beginning of the third millennium in the world of instrumental and laboratory diagnostics, in case of certain illnesses--so in the case of acute abdomen- physical examinations have not lost a bit from their significance. PMID- 11865757 TI - [Cacao in traditional Central American medicine]. PMID- 11865758 TI - [Authentic data to the biography of the dental surgeon Lajos Mathe of Bikafalva (1864-1917)]. PMID- 11865759 TI - Recommended core collection of Web sites for hospital libraries. AB - Core lists of recommended resources have long been used as selection tools for collection development in hospital libraries. However, hospital librarians now rely on the Internet for access to up-to-date information to supplement materials available within their libraries. Many libraries now either maintain a library Web page or provide input for their institution's Web page. The difficulty is determining what sites to include on their page. This article is a basic guide to recommended links for hospital libraries. It is intended as a starting place for hospital librarians who wish to create (or add to) a Web page of good links to be used for answering reference questions, or as a list for library users. The list is available as a link from the Sharp Health Science Library Web page where it will be updated at least annually. Links will be checked on a regular basis. PMID- 11865760 TI - Do you Mini-Med School? Leveraging library resources to improve Internet consumer health information literacy. AB - Popular for engaging public interest in medical science while promoting health awareness, Mini-Med School (MMS) programs also afford important if largely unrealized opportunities to improve the health information literacy of attendees. With a growing population using the Internet to make health decisions, needed venues for improving Internet Consumer Health Information (CHI) literacy may be found in the MMS platform. Surveyed directors of MMS programs understand the need to include CHI, and successful programs at SUNY Stony Brook and elsewhere demonstrate the potential for collaboration with affiliated health sciences libraries to integrate CHI instruction into MMS curricula. PMID- 11865761 TI - Literature search improvement project. AB - This paper describes an ongoing improvement effort directed at increasing the quality of mediated searches at the Sladen Library and Center for Health Information Resources. The project is the result of an analysis of literature statistics for mediated searching for 1997. The improvement project utilizes Deming's Plan-Do-Check-Act or PDCA cycle. Henry Ford Health System encourages use of the PDCA methodology for improvement projects. A key component of this improvement effort was the introduction of a productivity standard that each searcher is required to meet. The library has global productivity goals, but this is the first time that individual searchers have been held to a quantitative performance standard. The outcome of the Literature Search Improvement Project has been favorable. PMID- 11865762 TI - Decision support with Ovid. PMID- 11865763 TI - Reference uses for OCLC's Cooperative Online Resource Catalog (CORC): it's not just for catalogers. PMID- 11865764 TI - S.C.A.L.E.--spiritual care at life's end: a multi-disciplinary approach to end-of life issues in a hospital setting. AB - End-of-life care is a hot topic in many hospitals of late, in part due to an aging society, which is forcing health care professionals to address the issue. A culture that places enormous value on youth and avoids the topic of death also contributes to the discomfort many health care professionals feel discussing this topic with their patients and family members. Inpatient satisfaction survey scores at Thunderbird Samaritan Medical Center (TSMC) were very inconsistent on the question of how the institution was meeting the emotional and/or spiritual needs of patients. A staff survey and needs assessment was also conducted regarding end-of-life care, which pointed out communication issues among staff members. All of these factors led to the development of an inter-disciplinary team to examine the issue of end-of-life care at TSMC. PMID- 11865765 TI - Training opportunities for librarians sponsored by the National Library of Medicine. PMID- 11865766 TI - Genetically modified feeds in animal nutrition. 1st communication: Bacillus thuringiensis (Bt) corn in poultry, pig and ruminant nutrition. AB - During the last few years, animal nutrition has been confronted with genetically modified organisms (GMO), and their significance will increase in the future. The study presents investigations on the substantial equivalence of the transgenic Bt (Bacillus thuringiensis) corn and the corresponding nontransgenic hybrid Cesar and parameters of nutrition physiology such as digestibility and energy content for poultry, pigs and ruminants. The results of the analysed corn samples as well as of the silage samples illustrated substantial equivalence in all investigated ingredients, such as crude nutrients, amino acids, fatty acids, minerals and non starch polysaccharides. The results of the experiments using poultry, pigs, wethers and fattening bulls were not influenced by the genetic modification of corn. The determined values for the digestibilities and the energy contents for poultry, pigs and wethers were not affected by the used corn variety. Neither the examined parameters of the fattening experiments with bulls nor the slaughter results showed any significant differences between the bulls fed on silages made from the nontransgenic or transgenic corn. PMID- 11865767 TI - Genetically modified feeds in animal nutrition. 2nd communication: glufosinate tolerant sugar beets (roots and silage) and maize grains for ruminants and pigs. AB - To analyse substantial equivalence of genetically modified sugar-beets and maize, in which the glufosinate-tolerant (Pat) gene is inserted, crude nutrients, the amino acid and the fatty acid profiles as well as the composition of the NDF fraction of maize grains were determined and compared with those of the corresponding non-transgenic cultivars. Due to the genetic manipulation differences in crude nutrient contents including sugar and starch were not detected. The amino acid profile of maize grains was analysed to be the same. Fatty acid profile and composition of cell wall constituents did not show any influences as well. Digestibility of Pat-sugar-beets and maize grains for pigs did not demonstrate meaningful differences as compared to the corresponding non transgenic cultivars. Digestibility of sugar-beet roots and sugar-beet top silage for ruminants proved to be also in the scope of natural variance. As the digestibility of the macro nutrients remained unaffected, the Pat-gene introduction into both crops did not show an influence on the energetic feeding value. For pigs the ME-content of Pat-sugar-beets was determined to be 14.1 MJ/kg DM versus 13.7 MJ of the non-transgenic cultivars. ME-content of Pat-maize grains was 16.0 MJ/kg DM versus 15.8 MJ for controls. For ruminants the feeding value of Pat-sugar-beets was found to be 8.5 MJ NEL/kg DM or 13.2 MJ ME/kg DM, regardless of whether the Pat-gene was inserted or not. The corresponding energy values of sugar-beet top silage ranged between 5.2 and 5.5 MJ NEL/kg DM or 8.6 and 9.1 MJ ME/kg DM, with differences considered in the biological range. PMID- 11865768 TI - Endogenous N-losses in piglets estimated by a [15N]-isotope dilution technique: effect of xylanase addition to a wheat and rye based diet. AB - The nitrogen pool of piglets weighing 19.4 +/- 1.4 kg at the beginning of the experiment was labeled with an oral application of ([15N]H4)2SO4 (1.26 [15N]-atom percent excess of dietary N) over a period of 7 d. The labeling period was followed by an equilibration period of 7 d without feeding the labeling compound. The two experimental diets were based on wheat (53%) and rye (25%) and were fed either with or without a xylanase containing enzyme preparation over both experimental periods. Additionally, diets were supplemented with an indigestible marker during the 2nd period of the experiment to allow the calculation of endogenous N-losses in subsequent segments of the digestive tract of the pigs. These endogenous N-losses were estimated at the end of the experiment by analyzing feces, ingesta and urine for [15N]-enrichment assuming that [15N] enrichment of urine represents the [15N]-enrichment of the precursor pool. Endogenous N-losses were not significantly affected by xylanase addition at any measurement site (stomach, 3 sections of the small intestine, total digestive tract). Endogenous N-proportions of total nitrogen amounted on average for the six pigs to 42 +/- 11% and 56 +/- 5% at the last section of the small intestine and over the whole digestive tract, respectively, which corresponded to endogenous N-losses of 2.8 +/- 1.3 g N/kg DM and 2.0 +/- 0.3 g N/kg DM, respectively. PMID- 11865769 TI - Comparative investigation of salinomycin and flavophospholipol in sheep fed different composed diets. AB - The effects of salinomycin and flavophospholipol, and their relationship with the diet, were studied in nine ruminally and duodenally cannulated wethers. Within the composition of the ration, the levels of rumen degradable protein (RDP) and non-structural carbohydrates (NSC) were changed (diet H: 74% RDP and 38% NSC; diet M: 57% RDP and 32% NSC; diet L: 48% RDP and 23% NSC). There was no clear treatment effect of flavophospholipol on propionate concentration. Salinomycin supplementation appeared to be more effective than flavophospholipol in the increase of propionate concentration at the expense of acetic acid. Salinomycin significantly reduced the ammonia concentration of the rumen fluid. Microbial N content of the duodenal digesta was significantly lower when salinomycin was used. Salinomycin inhibited proteolysis and reduced the efficiency of microbial protein synthesis. The effect of salinomycin on ruminal N metabolism was independent of the composition of substrate. Unlike salinomycin, flavophospholipol tended to increase proteolysis in the rumen and did not inhibit protein synthesis. The effect of salinomycin on ruminal fermentation and the duodenal flow of nutrients were independent of substrate composition. PMID- 11865770 TI - Influence of hay particle size at different concentrate and feeding levels on digestive processes and feed intake in ruminants. 2. Passage, digestibility and feed intake. AB - In order to study the main effects of particle size, three ruminally fistulated cows (550 to 580 kg BW) were fed a constant low concentrate level (3.56 kg DM/d, 20% of total DMI) and a fibre-rich hay (approximately 60% NDF in DM) in long (28.7 mm), chopped (9.2 mm) and fine ground (2.9 mm) form in a 3 x 3 Latin square design. In another three factorial experiment with 8 wethers (4 animals were ruminally fistulated, mean BW = 68 kg) the main effects and interactions of the above mentioned hay particle sizes at two concentrate levels (10.4 to 13.3% and 29.5 to 40.1% of DMI, resp.) and two intake levels (restricted and ad libitum) were investigated. In comparison to long hay (28.7 mm), feeding of chopped hay (9.2 mm) at low concentrate levels, increased not only the hay intake (7% in dairy cows and 13% in sheep) but also the intake of digestible organic matter (12% in dairy cows and 32% in sheep), due to an increase in the apparent digestibility of OM by 3.8% in dairy cows and 8.2% in sheep. Ad libitum feeding of fine ground hay in combination with low concentrate amount in the ration increased the passage rate in the hindgut and consequently the hay intake, but not the intake of DOM, due to a significant depression of digestibility, especially of fibre fractions (4 to 7% in dairy cows and 4.5 to 14% in sheep), in comparison to 28.7 and 9.2 mm hay particle sizes. The digestibility decreased significantly with restricted feeding of fine ground hay in sheep only in comparison to 9.2 mm particle size. A threefold increase of concentrate amount leveled out all effects of the particle size reduction. The effect of particle size was more pronounced in sheep than in dairy cows. PMID- 11865771 TI - Research note: effect of duration of the collection period on the estimated digestibility in dairy cows. AB - A digestion trial was performed with 14 lactating Friesian dairy cows to compare the estimated digestibility of crude nutrients after collecting the faeces for 5, 7 and 9 d. The experimental ration consisted of grass silage (7.3 kg DM) and concentrate (10.9 kg DM). Animals were adapted to the ration for 14 d. There was no effect of the duration of the collection period on the estimated mean digestibility of crude nutrients and on the respective standard deviation. In consequence, in digestibility trials with cows the collection period must not exceed 5 d. PMID- 11865772 TI - [What is your diagnosis? Dorsal cortical triquetum fracture]. PMID- 11865773 TI - [Effect of aerobic endurance exercise on immune function in elderly athletes]. AB - Recently, there have been several reports on the influence of physical activity on immune function. Most of the studies were conducted using maximal exercise in young and middle-aged subjects. Since human immune function undergoes adverse changes with aging, we investigated in the present study whether submaximal, aerobic exercise induces changes in immune function in elderly subjects. Leukocytes, differential blood count, subsets of lymphocytes, CD4/CD8 ratio and immunoglobulins were studied after submaximal aerobic exercise (mean lactate 2.57 + 0.3 mmol/l) in 15 elderly subjects (mean age 68 +/- 5.6 years). These parameters were measured before, immediately after and 4 hours after exercise. Mean hemoglobin was unchanged indicating no hemoconcentration. There was a small increase in mean total lymphocytes (p < 0.05 immediately after exercise), while there was a highly significant increase in leukocyte count both immediately and 4 hours after exercise (p < 0.01). We found a significant correlation between the increase in leukocytes and lactate concentration (p < 0.01, r = 0.784). Lactate levels of all subjects were below 4 mmol/l. These results might indicate that the effect of a single bout of aerobic exercise on immune function depends on the intensity and duration of exercise relative to the level of fitness in elderly subjects. There was a highly significant rise (p < 0.001) in CD4/CD8 ratio 4 hours after exercise. This increase was mainly due to a rise in CD4 cell number whereas T-suppressor cells were almost unchanged. Our data show a possible stimulation of immune function by a submaximal, aerobic exercise in elderly subjects. Further studies are required to clarify whether a stimulation of CD4/CD8 ratio by chronic training gives rise to a reduction of infections in physically active elderly persons. PMID- 11865774 TI - [Suspected neurological side-effects of tick-borne meningoencephalitis vaccination: experiences of the Swiss Adverse Drug Reaction Reporting Center]. AB - The number of patients affected by tick-born encephalitis (TBE) in Switzerland has increased in the last years and an extension of the endemic foci of TBE has been observed. Therefore, active immunization by TBE vaccination has become more important. The possible adverse vaccine reactions have to be known as exactly as possible. The Swiss Drug Monitoring Center SANZ received from 1987 until June 2000 33 spontaneous cases reporting on 39 neurological adverse reactions in a close temporal relationship with a TBE vaccination and a suspected causal relationship. The following adverse reactions were reported most frequently: headache in 36%, neuropathy in 18% and meningeal irritation in 13%. Twelve out of 33 patients were hospitalized due to the adverse reaction. All neurological reactions were reversible. The spontaneous reporting scheme of the SANZ does not allow to calculate the incidence of neurological reactions after TBE vaccination. In general, adverse neurological reactions after TBE vaccination seem to be rare. According to the experiences of SANZ all reported neurological reactions were reversible. PMID- 11865775 TI - [Clinical aspects, etiology, pathogenesis, diagnosis and therapy of aseptic bone necrosis--a current analysis of the literature]. AB - With continuous proliferation of magnetic resonance imaging early detection of avascular bone necrosis becomes progressively important for treatment of early disease stages. The last decades intensified research on the field of ostoenecrosis has lead to different etiopathogenetic models and to new treatment options. Thereby long term results of known operative interventions are now available. Those can be used as a standard of treatment, new innovative techniques (e.g. mosaic arthoplasty or chondrocyte transplantation) might also be added. The goal of this paper is to give a comprehensive review about Osteonecrosis, Osteochondrosis and Osteochondritis dissecans. Respectively the latest literature results we are able to subordinate these to one unit with mechanical stress as a significant etiological factor in common. Knowledge derived form single literature marks are perhaps comprehended to general picture. PMID- 11865776 TI - [New prospects in diagnosis and treatment of heart failure]. PMID- 11865777 TI - [25-year-old medical assistant, post-exposure HIV prevention after accidental needlestick injury]. PMID- 11865778 TI - Definition of a correlation between microbiological and physical particulate barrier performances for porous medical packaging materials. AB - An experimental set up, assembled principally from components of commercially available filter test equipment, has allowed assessment of physical particulate barrier performance of porous materials. Employing twelve specially made papers with graded and widely different barrier properties, a physical particulate/microbiological correlation has been derived covering the 50,000 fold range of microbiological barrier performances seen in commercial porous medical packaging materials. The derived correlation has been shown to be independent of sheet structure and is applicable to the diverse structures that comprise commercial materials. A simple measurement of particle penetration is proposed as a means of assessing microbial barrier properties of such materials. PMID- 11865779 TI - Pall Corporation advocates the "broad-scale substitution". PMID- 11865780 TI - Study of protein adsorption effects on crossflow filtration using BSA and milk protein. AB - Three membrane materials were tested under similar conditions to determine the effects of membrane material on the performance and cleanability of the filters. The membrane materials investigated were stabilized cellulose (Hydrosart, Sartorius Corporation), cellulose triacetate, and polyethersulfone; all having a 10 kilodalton molecular weight cutoff. Stabilized cellulose is a cellulose-based membrane material, modified for low non-specific protein adsorption combined with high pH-resistance. When analyzing the data, three phenomena were considered: adsorption of protein to the membrane, gel layer formation, and osmotic pressure due to concentration polarization. Throughout these studies, the effects of a gel layer and the osmotic pressure were approximately equivalent in all cassettes. However, the stabilized cellulose was resistant to protein adsorption while the other two membranes exhibited significant decreases in permeate flux due to adsorption. Using a 0.2% BSA (MW = 67,000 kD) solution with a crossflow rate of 5 L/min and transmembrane pressure of 35 psi, the permeate flux through the stabilized cellulose membrane was 3% lower than the baseline saline flux, whereas the cellulose triacetate and polyethersulfone membranes lost 33% and 60% of baseline flux, respectively. The decrease in flux occurring in the latter two membranes is due to adsorption. Another study evaluated adsorption by alternating between crossflow and static operation. After three cycles, the stabilized cellulose maintained the original crossflow flux level. The polyethersulfone lost cumulatively 17% of its crossflow flux after three cycles and the cellulose triacetate lost 13%. The stabilized cellulose and polyethersulfone membranes were also evaluated using a milk solution. The results indicate that the stabilized cellulose is not susceptible to adsorption of any of the milk components while the polyethersulfone permeate flux was limited by adsorption. The saline flux of the stabilized cellulose immediately after testing with the milk solution was 3% lower than the baseline flux, while the polyethersulfone membrane saline flux was 81% lower. The results consistently indicated that, unlike the cellulose triacetate and polyethersulfone membranes, the stabilized cellulose membrane was not subject to adsorption. PMID- 11865781 TI - "Visually clean" as a sole acceptance criterion for cleaning validation protocols. AB - The role of visual examination as the sole acceptance criterion in cleaning validation protocols is explored, including the proper definition of "visual limit" for a given residue. Such a visual limit is specific to the surface the residue is on, and is further defined by viewing conditions, such as lighting, distance, and angle of viewing. A visually clean standard may only be properly utilized if the visual limit is below any scientifically calculated residue limit, such as that determined by a traditional dose-based limit determination. While such an approach, properly applied, has scientific justification and appears to be accepted by the proposed Annex 15 to the European GMPs, the approach is still untested with regulatory authorities. PMID- 11865782 TI - Review of standard for evaluating the effectiveness of contact lens disinfectants. AB - A new standard has been developed by the International Organization for Standardization (ISO) for evaluating contact lens disinfectants. This paper reviews the ISO 14729 standard and applies it to marketed products. The historical significance, justifications, scope, interpretations, application, and worthiness are discussed. The standard provides a more consistent procedure and a higher standard for contact lens disinfectants. As a result, more effective contact lens disinfectants have been marketed. PMID- 11865783 TI - Considerations in sterile filtration--Part I: The changed role of filter integrity testing. PMID- 11865784 TI - [Delayed diagnosis of benign paroxysmal vertigo: economic impact]. AB - Some time has passed since the early 1980's when a group of vestibologists with the same singular passion introduced in Italy the concept of Paroxysmal Positional Vertigo (PPV). Since then great advances have been made, but, despite the energy focused on improving and expanding the knowledge on this common disorder, PPV still frequently goes unrecognized. The direct consequence of delayed diagnosis is prolonged patient discomfort and the execution of useless, costly clinical examinations. Today, in most cases, delayed diagnosis is unjustified. Within the modern concept of health care economics, such delay is costly to the health care provider and has obvious psycho-physical repercussions for the patient who has to undergo a fruitless series of examinations and treatments of all types without reaping any benefits. In this light the Authors have performed a study to identify and quantify the relative costs of the most common errors made in the diagnosis of PPV. For a sample population of 100 patients, the clinical-instrumental tests performed before the disorder was correctly diagnosed were gathered and analyzed. Then the costs for the management of PPV patients diagnosed late were compared with those for patients whose diagnosis was reached early on. The results indicate that for each delay in PPV diagnosis the average cost was Euro 317.66 vs. Euro 64.04 for early diagnosis: a difference of Euro 253.06. The Authors underline how in this brief period of time (March-September 2000) the prescription of excessive testing led to a significant waste of money (Euro 10137.01). PMID- 11865785 TI - [Non-auditory effects of environmental noise: a study of metallurgical and mechanical workers]. AB - The present work was performed to check the non-auditory effects of environmental noise on a homogeneous worker population. The study was performed on 200 workers (factory and office workers) in a metallurgical and mechanical company: each individual underwent thorough clinical-audiometric study and filled out a multiple-choice psycho-social-attitude questionnaire. Statistical analysis of the results obtained showed a significantly higher number of non-auditory effects in subjects exposed to noise vs. those not exposed. For all workers studied, this incidence increased proportionally with age and job seniority. Moreover, it was demonstrated that there is no direct correlation between non-auditory effects and chronic traumatic hearing loss. The limited number of cases examined, however, suggests a multi-center study in order to provide better clinical-scientific and statistical-epidemiological study of this complex question involving on-the-job noise. PMID- 11865786 TI - [Eutectic mixture of local anesthetics (EMLA): evaluation of the analgesic effectiveness during ND: YAG laser turbinoplasty]. AB - The analgesic effect of EMLA anesthetic cream is evaluated during contact turbinoplasty with a Neodymium: yttrium-aluminum-garnet (Nd: Yag) laser. A total of 48 patients were selected and randomized in two groups. EMLA was applied on the nasal mucosa, respectively, for 15 and 30 minutes after using vasoconstrictor cottonoids. Both groups underwent turbinate reduction by contact with Nd: Yag laser using three discharge incisions traced parallel on the medial surface of the lower turbinate working from the tail to the head. The pain induced by the surgical procedure was evaluated with a visual analogic scale. The score obtained was significantly lower in the 1st group than in the 2nd group and this difference was statistically significant (p < 0.01). The EMLA anesthetic cream applied on the mucosa for 15 minutes allows to obtain a good analgesia during the functional contact turbinate surgery with a Nd: Yag laser and thus can be considered a valid alternative to anesthesia by local infiltration. PMID- 11865788 TI - [Prosthetic rehabilitation with endosseous implants in fibular free flaps reconstructed mandibles]. AB - Mandibular defect following neoplasm resection involves severe alterations in morphology. Today, however, these alterations can be repaired with the use of revascularized free flaps. Chewing can be returned to normal, or at least to an acceptable level, thanks to the insertion of endosseous implants, used both to support the fixed prosthesis and the mobile prosthesis (overdenture). The present work reports the Authors' experience with 5 patients who underwent compound mandibular resection and reconstruction with revascularized fibula free flaps. In all cases the mandible was totally toothless and rehabilitation called for the insertion of a total prosthesis held by a bar secured to the endosseous implant. In 2 cases the implants were inserted at the time of reconstruction. Four patients completed the rehabilitation program, with a 93% (14/15) implant success rate. All were able to return to a normal diet. Likewise the aesthetic results were valid. The main problem was to achieve an adequate interface between the soft tissues and the implants. PMID- 11865787 TI - [Primary reconstruction of lateral oro-mandibular defects using titanium plates (Thorp) and revascularized non-osseous flaps: a personal experience]. AB - Reconstruction of a particular defect is an highly complex problem, that is reflected in the number and variety of techniques proposed. This is particularly true for reconstruction of mandibular segment defects usually together with the need to repair oro-pharnygeal soft tissues defects of various size. The high rate of successes that can be obtained today using microvascular reconstruction techniques explains the reason why bone free flaps have progressively become the "gold standard" used as benchmark for all other forms of primary mandibular reconstruction. However, for selected patients (elderly patients or individuals in poor general health; the presence of lateral oro-mandibular defects or soft tissue defects significantly exceeding the bone defect) the association of non osseous microvascular free flaps (such as forearm skin flaps or miocutaneous free flaps of the abdominal rectus muscle) with a titanium mandibular reconstruction plate (THORP) can prove a valid alternative. Indeed, in these cases reconstruction of choice would be the association of two free flaps, one osseous and the other non osseous. The use of THORP-type mandibular reconstruction plates in repairing lateral defects increases the overall risk of complications related to the reconstruction technique which is already hindered by the risks associated with the use of microvascular free flaps. The present work starts with a retrospective evaluation of 5 years series of primary mandibular reconstruction for lateral oro-mandibular defects following oncological surgery. The role of THORP-type titanium mandibular plates, used in combination with non-osseous free flaps in the treatment of selected patients, is then described and discussed. PMID- 11865789 TI - [Gastro-esophageal reflux and laryngeal phlogistic disorders: clinical evaluation and multi-electrode pH monitoring]. AB - Gastro-esophageal reflux (GER) is a complex pathology characterized by many clinical symptoms: gastroenterological (typical symptoms) and odontostomatological, pneumological and, more frequently, otorhinolaryngological (atypical symptoms). For this last manifestations no cause-effect relationship has yet been proven. Using a validated questionnaire about the typical reflux symptoms and the use of substances of abuse (tobacco and alcohol) and with the application of instrumental means such as electronic videolaryngoscopy and multi electrode 24 hours esophageal pH monitoring, the Authors have studied 34 selected patients (16 female, 18 male) suffering from chronic clinical laryngeal symptoms and presenting a peculiar pharyngeal-laryngeal objectivity. Both these manifestations could be ascribed to atypical GER symptoms. Analysis of the data showed that 67.6% of the subjects presented pathological reflux, with a significant prevalence of inflammatory lesions in the posterior portion of the larynx (in 47.8% laryngitis and retrocricoid edema). The validated questionnaire on typical reflux symptoms showed that 69.5% did not complain of the typical gastro-esophageal symptoms thus indicating that pharyngeal-esophageal reflux can be totally asymptomatic. While confirming the strong association between inflammatory pathologies and GER, not enough cases have been studied to identify a direct cause-effect relationship and this is complicated by the presence of other concomitant risk factors such as cigarette smoking or alcohol consumption. Therefore the Authors underline the importance of an in-depth, multi-discipline approach in the evaluation of pharyngeal-laryngeal phlogoses. This approach must make use of all available modern diagnostic techniques (Electronic videolaryngoscopy, multi-electrode pH monitoring) in order to obtain as much information as possible to throw light on the pathogenic mechanism responsible for the damage and find more effective forms of anti-reflux therapy. PMID- 11865790 TI - [Z-shaped surgical flaps in the treatment of acquired glottic stenosis: case report]. AB - The Authors report a case of laryngeal stenosis initially treated through cervicotomy with the application of an Eliachar prosthesis and finally resolved, after recurrence, with two Z-shaped mucosa flaps. The Authors feel that this method is worthy of note because: its is simple to perform; provides excellent flap vitality; makes a stent unnecessary; and, above all, provides positive results. PMID- 11865791 TI - [Dermoid cysts of the floor of the mouth: case report]. AB - Dermoid cysts are for the most part benign neoformations most likely deriving from ectodermic differentiation of totipotent cells. Although localization on the floor of the mouth has been described for more than a century it is still reported as rare and can give rise to problems in differential diagnosis. This work presents a case of dermoid cyst of the floor of the mouth in a 16-year-old patient and provides considerations on clinical approach, diagnosis and therapy for this type of lesion. PMID- 11865792 TI - [Acoustic neuroma: clinical-functional finding, results and surgical complication]. AB - The present work provides clinical-functional findings, results and surgical complications observed in a consecutive series of 100 subjects with acoustic neuroma (AN). Analysis of the data has made it possible to draw some important conclusions. Compromised hearing is found in 90% of the ears affected by AN. Indeed the percentage of normal hearing in such cases does not exceed 5%. There is, however, no clear correlation between degree of hearing and tumor size. The symptoms of AN do not always present unilateral or asymmetrical hearing loss, unilateral tinnitus and/or dizziness. At times AN presents atypical symptoms and can even be asymptomatic. Sudden onset of unilateral hearing loss, acute vertigo, persistent monolateral tinnitus and even isolated symptoms of the V or VI cranial nerve should lead one to suspect AN. Only by applying the diagnosis of suspected AN in a large number of cases is it possible to lower the time gap between the onset of symptoms and the definitive diagnosis of AN, increasing the number of cases diagnosed while the AN is still small. Auditory brainstem responses (ABR) are still the means of choice for screening and following up subjects where AN is suspected. Reduced ABR sensitivity reported in the literature for intracanal ANs must induce further testing with magnetic resonance imaging with gadolinium in all subjects where an AN is suspected, even when the ABR is normal. Recording of transient evoked otoacoustic emissions in the presence and in the absence of contralateral white noise has proved to be a simple, inexpensive, non-invasive test for the diagnosis of suspected retrocochlear pathologies. A deficit in vestibular function is most frequently encountered when the AN is already quite large and an alteration in the smooth pursuit test is only found when the AN involves the brainstem. These data have led us to conclude that vestibular reflex studies do not play any role in early diagnosis of AN. Surgical exeresis is the treatment of choice in those cases where "watch and scan" (only hearing ear in the absence of neurological complications; AN < 0.5 cm in the ponto-cerebellar angle, particularly in elderly patients) is not indicated. The enlarged translabyrinthine approach is indicated in all cases of AN, no matter what the tumor size and extent of pre-operative hearing. Promptly and correctly treating intra and postoperative complications, most frequently encountered in patients with AN > 2 cm, reduces the mortality and morbidity to a minimum. Modern otological microsurgery and monitoring techniques make it possible to preserve the VIIth facial nerve in more than 90% of the ears, consequently preserving or nearly preserving normal VIIth nerve function 1 year after surgery in at least three out of four patients. No matter what approach is used, hearing can be preserved measurably in approximately 50% of the ears undergoing surgery and to a socially useful or nearly useful level in a significantly lower proportion of patients. In this regard the most satisfactory results are obtained when preoperative hearing is normal and the AN is < 2 cm. PMID- 11865793 TI - The Youth Risk Behavior Surveillance System: updating policy and program applications. AB - To monitor behaviors that place adolescents at increased risk for premature morbidity and mortality, the Centers for Disease Control and Prevention developed the Youth Risk Behavior Surveillance System (YRBSS). This system measures six categories of behaviors, including behaviors that contribute to violence and unintentional injuries; tobacco use; alcohol and other drug use; sexual behaviors that contribute to unintended pregnancy and sexually transmitted diseases, including HIV infection; unhealthy dietary behaviors; and inadequate physical activity. This article summarizes how some education and health agencies and nongovernmental organizations, in collaboration with community agencies, school boards, parents, and youth, use YRBSS data to describe risk behaviors, create awareness, supplement staff development, set and monitor program goals, develop health education programs, support health-related legislation, and seek funding. Ways in which YRBSS data are distributed electronically also are summarized. PMID- 11865794 TI - Parent knowledge and opinions of school health services in an urban public school system. AB - Health services, provided through schools for more than 100 years, increasingly have expanded to meet preventive as well as acute health care needs of children. This article reports on a survey of parents of third-grade children in an urban public school system. The authors examined what parents know about school health services, what value they place on the services, and what barriers exist to health care access. Results indicated parents place a high value on health services offered in schools, but they know little about service availability and use. Parents often were unaware their children received many of the services listed, such as review of school health records, vision and hearing screening, and health education by school nurses. PMID- 11865795 TI - The Activated Health Education model: refinement and implications for school health education. AB - A behaviorally based, health education instructional model was developed from a review of successful health education interventions. The analysis identified three principles common to improvement of health behavior among program participants: 1) active involvement in the intervention process, 2) awareness of the positive and negative influences of the behavior on health, and 3) personal responsibility for managing health behavior. These principles and several theoretical premises then were organized into a structural framework for the model. Next, studies were conducted to validate applications of these conditions and refine strategic components. The model then was formatted into a curricular design to achieve optimum behavior and provide ease of replication. Research related to developing and field testing the model are included. The model's limitations and strengths, implications for school health education, and plans for the future are presented. PMID- 11865796 TI - Report of the 2000 Joint Committee on Health Education and Promotion Terminology. PMID- 11865797 TI - Environmental allergens and irritants in schools: a focus on asthma. AB - As part of the Partners in School Asthma Management Program, environmental data were collected from 385 rooms in 60 elementary schools in southeast Texas, using an Environmental Observation Checklist and a Q-TRAK Indoor Air Quality Monitor. Dust samples for allergen analysis were collected from floors, carpets, and area rugs in 80 classrooms in a subset of 20 schools. CO2 levels > 1,000 ppm were found in 86% of rooms; 69% had indoor humidity above recommended levels. Der p I dust mite allergen levels > 2,000 ng/g were present in 20% of rooms, but only 2.5% of rooms had Der f I mite allergen levels exceeding recommended tolerances. Detectable levels of cockroach allergen (Bla g II) were found in all schools (median 5.5 ng/g), with 10% of rooms over the recommended threshold. Almost two thirds of classrooms had mold spore counts > 10,000 col/g (median, 14,400 col/g; range, 2,000-52,000 col/g). PMID- 11865798 TI - The alarming increase of type 2 diabetes in children. PMID- 11865799 TI - "Tinkering" with the components of health. PMID- 11865800 TI - Opinions of Illinois voters about coordinated school health programs. AB - A random-digit dialed telephone survey was conducted in Illinois to determine support for coordinated school health programs (CSHP). Survey data were obtained from 469 registered voters in June and July 2000. Results demonstrated that most Illinois registered voters: 1) supported the concept and practice of CSHP; 2) were uncertain if current school health education and services were adequate; 3) believed Illinois schools should set a goal of creating a "healthier state;" and 4) although supportive of concept, were not familiar with term CSHP. Characteristics of supportive and nonsupportive registered voters are reported. Results of this study may assist in developing educational and marketing strategies to build support for coordinated school health programs in Illinois. PMID- 11865801 TI - Hemophilia Care in the New Millennium. Proceedings of a conference dedicated to Dr. Roberts. Chapel Hill, North Carolina, USA. April 8-10, 1999. PMID- 11865802 TI - Proceedings of the Hammersmith Perfusion Workshop: Perfusion haematology. December 2000. PMID- 11865803 TI - Proceedings of the 8th International Antiviral Symposium and Workshop. Kagoshima, Japan, 19-21 November 2000, in honour of Professor William Prusoff and Professor Shiro Shigeta. PMID- 11865804 TI - [Multiple Sclerosis. Consensus Conference Organized by the French Federation for Neurology. 7-8 June 2001]. PMID- 11865805 TI - Management of malignant pineal germ cell tumors with residual mature teratoma. PMID- 11865806 TI - Endoscopic endonasal cavernous sinus surgery: an anatomic study. PMID- 11865807 TI - High serum S100B levels for trauma patients without head injuries. PMID- 11865810 TI - Salary survey. Clinical laboratory personnel. PMID- 11865809 TI - Challenges associated with chronic medication compliance and adherence. PMID- 11865811 TI - First meeting of the Gene Vector Production network, 4-5 October 2001, Evry, France. PMID- 11865812 TI - A question that won't go away. PMID- 11865813 TI - Gemcitabine in metastatic nasopharyngeal carcinoma of the undifferentiated type. AB - BACKGROUND: We conducted two parallel phase II trials in chemonaive and previously treated patients with metastatic nasopharyngeal carcinoma (NPC) to evaluate the tumour response, progression-free and overall survival, and toxicity of gemcitabine. PATIENTS AND METHODS: Gemcitabine 1250 mg/m2 was given on days 1 and 8 of a 21-day cycle. Patients with an Eastern Cooperative Oncology Group performance status <2, adequate renal, hepatic and bone marrow function, and radiologically measurable NPC were eligible. RESULTS: Twenty-five chemonaive and 27 previously treated patients were enrolled. The overall response rate was 28% [95% confidence interval (CI) 14% to 48%] for the chemonaive and 48% (95% CI 31% to 66%) for previously treated patients. Toxicities greater than or equal to grade 3 occurred in 15 (60%) chemonaive and 13 (48%) previously treated patients. Neutropenia was uncommon in chemonaive patients, but occurred in 37% of previously treated patients. The median time to progression was 3.6 months (range 0.9-7.9) for chemonaive and 5.1 months (0.9-13.1) for previously treated patients. Median overall survival time was 7.2 months (1.4-15.6) and 10.5 months (2.4-15.0) for chemonaive and previously treated patients, respectively. CONCLUSIONS: Gemcitabine has moderate activity in NPC with minimal toxicity, and is also an effective salvage agent for patients who have failed or progressed after treatment with other agents. PMID- 11865814 TI - Trimetrexate as biochemical modulator of 5-fluorouracil/leucovorin in advanced colorectal cancer: final results of a randomised European study. AB - BACKGROUND: Trimetrexate (TMTX) is a biochemical modulator of 5-fluorouracil (5 FU) and leucovorin (LV). Phase II trials have shown promising activity of 5 FU/LV/TMTX in patients with advanced colorectal cancer (ACC). This trial evaluated the effect of TMTX in combination with 5-FU/LV as first-line treatment in ACC. PATIENTS AND METHODS: Patients with ACC were randomised to receive either intravenous LV 200 mg/m2/5-FU 600 mg/m2 or TMTX 110 mg/m2 followed 24 h later by LV 200 mg/m2/5-FU 500 mg/m2 plus oral LV rescue. Both schedules were given weekly for 6 weeks every 8 weeks. Patients were evaluated for progression-free survival (PFS), overall survival (OS), tumour response, quality of life (QoL) and toxicity. RESULTS: A total of 365 patients were randomised. A statistically significant prolongation of median PFS was seen in patients treated with TMTX/5 FU/LV compared with 5-FU/LV (5.4 months versus 4.1 months, respectively; P = 0.03), and a trend towards a significant benefit for OS (13.4 months versus 10.5 months, respectively; P = 0.08). Tumour response, QoL and toxicity were comparable between the two arms. Diarrhoea was the most frequently occurring grade 3 or 4 toxicity (22% and 30%, respectively). CONCLUSIONS: The addition of TMTX to a weekly regimen of 5-FU/LV results in a small but significant improvement in PFS without adding toxicity or worsening QoL in patients with ACC. PMID- 11865815 TI - Integrated analysis of overall survival in two randomised studies comparing 5 fluorouracil/leucovorin with or without trimetrexate in advanced colorectal cancer. AB - BACKGROUND: Two randomised studies were performed with trimetrexate (TMTX) as a biochemical modulator of 5-fluorouracil (5-FU)/leucovorin (LV) in advanced colorectal cancer (ACC), one in Europe and one in the United States. Both studies were similarly designed to detect a statistically significant difference in progression-free survival (PFS). Overall survival (OS), however, was later adopted as the primary outcome measure for approvability of agents for first-line treatment of ACC. Therefore, an integrated analysis of survival data from the European and USA trials was performed to detect a clinically relevant difference in survival. PATIENTS AND METHODS: The experimental arm was identical in both studies and consisted of TMTX 110 mg/m2 intravenously (i.v.) followed 24 h later by i.v. LV 200 mg/m2/5-FU 500 mg/m2 plus oral LV rescue. The 5-FU dose in the control arm was 600 mg/m2 in the European study and 500 mg/m2 in the USA study, and the USA study was placebo-controlled. Treatment was given weekly for 6 weeks every 8 weeks. RESULTS: A total of 746 patients were analysed. Median OS was 13.0 months for 5-FU/LV and 14.6 months for TMTX/5-FU/LV (P = 0.15; Wilcoxon). Median PFS was 4.4 months and 5.4 months, respectively (P = 0.07; Wilcoxon). CONCLUSIONS: The addition of TMTX to a weekly regimen of 5-FU/LV does not improve the outcome for patients with ACC. PMID- 11865816 TI - Source levels of clicks from free-ranging white-beaked dolphins (Lagenorhynchus albirostris Gray 1846) recorded in Icelandic waters. AB - This study reports the source levels of clicks recorded from free-ranging white beaked dolphins (Lagenorhynchus albirostris Gray 1846). A four-hydrophone array was used to obtain sound recordings. The hydrophone signals were digitized on line and stored in a portable computer. An underwater video camera was used to visualize dolphins to help identify on-axis recordings. The range to a dolphin was calculated from differences in arrival times of clicks at the four hydrophones, allowing for calculations of source levels. Source levels in a single click train varied from 194 to 211 dB peak-to-peak (p-p) re: 1 microPa. The source levels varied linearly with the log of range. The maximum source levels recorded were 219 dB (p-p) re: 1 microPa. PMID- 11865817 TI - A wide angle and high Mach number parabolic equation. AB - Various parabolic equations for advected acoustic waves have been derived based on the assumptions of small Mach number and narrow propagation angles, which are of limited validity in atmospheric acoustics. A parabolic equation solution that does not require these assumptions is derived in the weak shear limit, which is appropriate for frequencies of about 0.1 Hz and above for atmospheric acoustics. When the variables are scaled appropriately in this limit, terms involving derivatives of the sound speed, density, and wind speed are small but can have significant cumulative effects. To obtain a solution that is valid at large distances from the source, it is necessary to account for linear terms in the first derivatives of these quantities [A. D. Pierce, J. Acoust. Soc. Am. 87, 2292 2299 (1990)]. This approach is used to obtain a scalar wave equation for advected waves. Since this equation contains two depth operators that do not commute with each other, it does not readily factor into outgoing and incoming solutions. An approximate factorization is obtained that is correct to first order in the commutator of the depth operators. PMID- 11865818 TI - Acoustic radiation impedance of rectangular pistons on prolate spheroids. AB - The self and mutual radiation impedances for rectangular piston(s) arbitrarily located on a rigid prolate spheroidal baffle are formulated. The pistons are assumed to vibrate with uniform normal velocity and the solution is expressed in terms of a modal series representation in spheroidal eigenfunctions. The prolate spheroidal wave functions are obtained using computer programs that have been recently developed to provide accurate values of the wave functions at high frequencies. Results for the normalized self and mutual radiation resistance and reactance are presented over a wide frequency range for different piston sizes and spheroid shapes. PMID- 11865819 TI - So you want to be a specialist registrar?--What to put in your CV. AB - Dentists applying to a specialist training programme often receive conflicting advice over what to put in their curriculum vitae (CV). We conducted a survey of the Training Programme Directors of the dental specialties to determine what aspects of CV content and presentation styles are considered important. This has allowed us to construct guidelines for what to put in a CV. Recently, structured application forms have become increasingly popular and may be a more objective way to carry out the shortlisting process. The guidelines presented could also be used as a framework for medical personnel departments if structured application forms eventually replace the CV. PMID- 11865820 TI - Changing patterns and the need for quality. AB - This series of articles is aimed at anybody who places crowns and other extra coronal restorations (ie veneers and shims) on individual teeth. We hope that everyone from experienced practitioners to undergraduate students may find something of value. Whoever reads them, we would ask to do so with an open mind. We have tried not to be dogmatic, and the techniques and materials described are not the only ones available, but are the ones which accord with the principles we describe. PMID- 11865821 TI - Type I membranoproliferative glomerulonephritis in an HIV-infected individual without hepatitis C co-infection. AB - Type I membranoproliferative glomerulonephritis (MPGN) is an uncommon manifestation of human immunodeficiency virus (HIV)-associated renal disease in patients co-infected with hepatitis C virus (HCV). We describe a case of Type I MPGN in an HIV-positive diabetic man with nephrotic-range proteinuria and renal insufficiency who was not co-infected with HCV. Tubuloreticular inclusions were present but there was no evidence for either cryoglobulinemia or cryoglobulin deposits in the kidney. This finding suggests that Type I MPGN may represent a reaction of the kidney to HIV independent of the effects of HCV co-infection. Clinical suspicion must be maintained for Type I MPGN in all HIV infected patients presenting with significant proteinuria regardless of HCV infection status. PMID- 11865822 TI - Focal and segmental glomerulosclerosis and non-Hodgkin's lymphoma. PMID- 11865823 TI - Developmental risk factors for sexual offending. AB - OBJECTIVE: The aim of the study was to identify the general, common, and specific developmental risk factors for pedophilia, exhibitionism, rape, and multiple paraphilia, and to address five methodological issues observed in this area of research. METHOD: This study involved 64 sex offenders and 33 nonsex, nondrug related, and nonviolent property offenders. The group of 64 sex offenders was further divided into eight subgroups, some of which overlapped in memberships because of multiple diagnoses. To overcome the methodological problem associated with overlapping group memberships, a special approach involving comparisons of sets of logistic regression analyses was adopted. Offenders were clinically assessed for evidence of paraphilias, and their adverse childhood experiences were measured by a battery of tests. RESULTS: Childhood Emotional Abuse and Family Dysfunction, Childhood Behavior Problems, and Childhood Sexual Abuse were found to be general developmental risk factors for paraphilias. Furthermore, Childhood Emotional Abuse and Family Dysfunction was found to be a common developmental risk factor for pedophilia, exhibitionism, rape, or multiple paraphilia. Additional analyses revealed that childhood emotional abuse contributed significantly as a common developmental risk factor compared to family dysfunction. Besides, Childhood Sexual Abuse was found to be a specific developmental risk factor for pedophilia. CONCLUSIONS: The study has supported the value of conceptualizing certain childhood adversities as developmental risk factors for paraphilic behaviors. The role of childhood emotional abuse as an important developmental risk contributor, and the relationship between childhood sexual abuse and pedophilia are of theoretical significance. Furthermore, the results have significant implications for the prevention of childhood abuse and treatment of sex offenders. PMID- 11865824 TI - Biochemical properties and mechanism of action of a vanadyl(IV)-aspirin complex on bone cell lines in culture. AB - A recently synthesized vanadyl(IV) complex with aspirin [VO(aspirin)ClH2O]2, has been thoroughly investigated by physicochemical techniques. In order to support the proposed structure, stoichiometry and the coordination sphere of the vanadium center, some studies such as elemental analysis, electronic (diffuse reflectance) and vibrational (infrared) spectroscopies, magnetic susceptibility, as well as the thermal behavior, were carried out. The bioactivity of the vanadium complex (VOAspi) was evaluated on two osteoblast-like cell lines in culture, being its cytotoxic effects stronger than the vanadyl cation as assessed by morphological changes and lipid peroxidation. These effects may be partially explained through the induction of the expression of Erks (Extracellular signal-regulated kinases) and the inhibition of the PTPases (Phosphotyrosine phosphatases) present in the cellular extracts. PMID- 11865825 TI - Hemodynamic consequences of neuropeptide Y-induced obesity. AB - BACKGROUND: Acute central nervous system administration of neuropeptide Y (NPY) elicits variable hemodynamic responses. Chronic intracerebroventricular (ICV) administration of NPY produces obesity in rats. Obesity has been shown to increase arterial pressure. METHODS: In this study we examined the chronic hemodynamic effects of NPY-induced obesity. Sprague-Dawley rats were implanted with radiotelemetry transmitters to continuously record heart rate and arterial pressure in the conscious state. Neuropeptide Y or vehicle was delivered into the third cerebral ventricle by osmotic minipumps over 2 weeks. Three groups were studied: vehicle, NPY-treated (free-fed), and NPY-treated (pair-fed to vehicle treated rats). RESULTS: Neuropeptide Y increased food intake and body weight in free-fed animals, and substantially augmented visceral adiposity in both free- and pair-fed rats. Despite increased adiposity, chronic ICV administration of NPY in conscious unstressed rats did not increase arterial pressure. Neuropeptide Y decreased heart rate, suggesting a sympathoinhibitory effect. CONCLUSIONS: Obesity induced by 2-week ICV administration of NPY does not increase arterial pressure, perhaps indicating inhibition of sympathetic outflow that may oppose the pressor effect of adiposity. PMID- 11865827 TI - Canadian Digestive Diseases Week. Abstracts of the 41st annual meeting of the Canadian Association of Gastroenterology in conjunction with the Canadian Association for the Study of the Liver. January 30-February 5, 2002. Montreal, Quebec, Canada. PMID- 11865826 TI - Selective high-performance liquid chromatographic assay for itraconazole and hydroxyitraconazole in plasma from human immunodeficiency virus-infected patients. AB - A sensitive and selective reversed-phase liquid chromatographic assay for itraconazole and hydroxyitraconazole in human plasma has been developed and validated. Itraconazole and hydroxyitraconazole were extracted from the matrix using solid-phase extraction on a strong cation-exchange sorbent. All compounds were detected using fluorescence at 265 and 363 nm for excitation and emission, respectively. The assay has been validated over the range 10-1,000 ng/ml for both compounds, 10 ng/ml being the lower limit of quantification. Accuracies ranged from 104 to 113% for itraconazole and from 91 to 103% for hydroxyitraconazole. The intra-assay precisions were all below 9% for itraconazole and below 8% for hydroxyitraconazole. The selectivity has been evaluated with respect to all registered anti-human immunodeficiency virus (HIV) drugs and other potential co medications and a few of their metabolites, commonly used by HIV-infected individuals. Both itraconazole and hydroxyitraconazole were stable under relevant conditions for HIV-inactivation and storage of samples. The applicability of the assay was demonstrated for samples collected from a treated HIV-infected patient. PMID- 11865829 TI - Treatment of long-distance intermittent claudication with pentoxifylline: a 12 month, randomized trial. AB - The efficacy, safety, and cost of pentoxifylline (PXF) in long-range (>400 m interval) intermittent claudication was studied comparing PXF and placebo in a 12 month study. A standardized treadmill test was performed at inclusion and at 6 and 12 months. A training plan based on walking was associated with the control of risk factor levels. Of the 194 included patients, 135 completed the study: 75 in the PXF group and 60 in the placebo group. There were 59 dropouts (due to low compliance). The authors observed a 148% increase in total walking distance (TWD) at 6 months with PXF (vs 110% with placebo; p<0.05); at 12 months, the increase was 170% with PXF (vs 131% with placebo; p<0.02). There was a 38% difference at 6 months and 39% at 12 months in favor of PXF. Treatment was well tolerated. In conclusion, PXF improved walking distance significantly better than placebo. PMID- 11865828 TI - Treatment of severe intermittent claudication with pentoxifylline: a 40-week, controlled, randomized trial. AB - The efficacy, safety, and cost of pentoxifylline (PXF) in the treatment of severe intermittent claudication were studied comparing PXF and placebo in a randomized 40-week study. A treadmill test was performed at inclusion and at the end of weeks 20 and 40. A progressive training plan and the control of risk factors (with antiplatelet treatment) were used in both groups. Of the 200 included patients, 178 completed the study: 88 in the PXF group and 90 in the placebo group. There were 22 dropouts. The two groups were comparable for age, sex distribution, and for the presence of risk factors and smoking. There was a significant increase in pain-free walking distance (PFWD) in both groups. The absolute and percent increase in PFWD was significantly greater in the PXF group (p<0.05). At 20 weeks, the increase was 360.5% in the PXF vs 252% in the placebo group. At 40 weeks, the increase was 386% in the PXF and 369% in the placebo group (p<0.02). Total walking distance (TWD) increased at 20 weeks (up to 254%) and up to 329% at 40 weeks. In the placebo groups the increase was 158% at 20 weeks and 183% at 40 weeks. The excess increase produced by PXF treatment was 30% at 20 weeks and 38% at 40 weeks (p<0.02). Unwanted effects treatment was well tolerated. No serious drug-related side effects were observed. In summary, between-group analysis favors PXF considering walking distance and costs. Results indicate good efficacy and tolerability. PMID- 11865830 TI - Treatment of vascular inner ear disease with pentoxifylline: a 4-week, controlled, randomized trial. AB - The efficacy of pentoxifylline (PXF) in vascular inner ear disease (VIED) was studied comparing PXF and placebo in a 4-week study; 60 patients with unilateral loss of hearing, vertigo, dizziness, tinnitus (analyzed with an analogue scale line), and cochlear flow reduction were included. The aim of the study was to study the effect of PXF (1800 mg/day) in VIED considering clinical outcome and cochlear flow. All patients completed the study. Improvement in cochlear flow (p<0.05) and a decrease in score in both groups were observed. The cochlear flow increase was 287.5% in the PXF group vs 168% in the placebo group (119.5% difference; p<0.02). There was a difference in score decrease (44.1% larger) in the PXF group (p<0.05). PXF was more effective considering flow and symptoms. PMID- 11865831 TI - Treatment of vascular inner ear disease in vascular patients with pentoxifylline: a controlled, randomized trial. AB - The efficacy of Pentoxifylline in vascular inner ear disease (VIED) was studied comparing PXF and placebo in a 4-week study; 40 patients with vascular disease and monolateral loss of hearing, vertigo, dizziness, tinnitus (analyzed with an analogue scale line), and cochlear flow reduction were included. The aims of the study were to study the effects of PXF (1600 mg daily) in VIED considering clinical outcome and cochlear flow. Of the 20 included patients, 19 completed the study. One dropout in the placebo group was due to low compliance. Intention-to treat analysis indicated an improvement in cochlear flow (p<0.05) and a decrease in score in both groups. The cochlear flow increase was 287.5% in the PXF group vs 168% in the placebo group (119.5% difference; p<0.02). There was a difference in score decrease (44.1% larger) in the PXF group (p<0.05).Between-group analysis favors PXF considering blood flow and symptoms. Results indicate good efficacy and tolerability of PXF in VIED. PMID- 11865832 TI - Short-range intermittent claudication and rest pain: microcirculatory effects of pentoxifylline in a randomized, controlled trial. AB - The efficacy of pentoxifylline (PXF) in severe intermittent claudication was studied comparing PXF and placebo. Patients were randomized into two treatment plans: PXF (2400 mg, four 600-mg tablets daily) or equivalent placebo was administered for 10 days. The exercise protocol associated with treatment was conducted under supervision. Skin flux (RF) was measured at rest and after 1 minute of exercise (AEF = after exercise flux; 3 km/hr, 12% inclination) with laser Doppler. PO2 and PCO2 were measured at the dorsum of the foot. All 20 included patients completed the study. The two groups were comparable. In the PXF group there was a significant increase in RF, AEF, and in PO2 (p<0.05); PCO2 was decreased (p<0.05). There were also changes in the placebo group, significantly lower than those observed in the PXF group (p<0.05). In conclusion high-dose PXF treatments improved all microcirculatory parameters in subjects with short-range claudication even with a short period of treatment. PMID- 11865833 TI - Treatment of vascular retinal disease with pentoxifylline: a controlled, randomized trial. AB - The aim of this study was to evaluate the effect of PXF (1800 mg daily) in patients with sudden loss of vision (SLV) in a 4-week trial, evaluating clinical outcome and retinal flow parameters. Inclusion criteria were SLV associated with thrombosis of the retinal artery; decrease in retinal blood flow (PSF: peak systolic flow; EDF: end-diastolic flow velocity) and asymmetry between the two retinal arteries (>40%) documented by duplex scanning. All 10 included patients completed the study. The groups were comparable. No side effects were reported. A significant improvement in flow velocity (p<0.05) and a decrease in analogue score in both groups were observed. PSF increase was 550% in the PXF group vs 288% in the placebo group (262% difference). EDF increase was 400% in the PXF group vs 200% in the placebo group (200% difference). There was a significant difference in the analogue score decrease (33.3% difference larger in the PXF group; p<0.05). In conclusion, PXF treatment improved retinal flow after retinal artery occlusion better than placebo and should be considered as an important option in this condition. PMID- 11865834 TI - Treatment of retinal vein thrombosis with pentoxifylline: a controlled, randomized trial. AB - The aim of this study was to evaluate PXF (pentoxifylline; 1600 mg daily vs placebo) in patients with retinal vein thrombosis (RVT) in a 4-week trial, evaluating clinical outcome and retinal flow. Inclusion criteria were sudden loss of vision (SLV); retinal vein thrombosis (RVT); decrease in retinal vein flow; asymmetry between retinal veins (>40%) documented by duplex scanning (retinal vein thrombosis flow = RVTF). All 18 included patients completed the study. The groups were comparable. No side effect was observed. An improvement in arterial flow (p<0.05) and a decrease in analogue score (p<0.05) were observed in both groups (due to the spontaneous evolution with partial thrombus lysis in 4 weeks). The increase in arterial flow (PSF and EDF) were greater (p<0.05) in the PXF group. The RVFV increase was better in the PXF group (350% increase vs 200% increase in the placebo group; p<0.05). There was a significant difference in the analogue score decrease (4 vs 7) in the PXF group (p<0.05). In conclusion, PXF improved retinal flow after RVT better than placebo. It should be considered as an important treatment option. PMID- 11865835 TI - Intermittent claudication in diabetics: treatment with exercise and pentoxifylline--a 6-month, controlled, randomized trial. AB - The aims of this study were to evaluate the effect of PXF (1600 mg daily) in diabetic patients with intermittent claudication. Of the 60 included patients, 53 completed the study (27 in the PXF group). There were seven dropouts. The groups were comparable for age, sex distribution, and total walking distance (TWD), and risk factors. There was an increase in TWD at 3 and 6 months in both groups (p<0.05) possibly due to exercise. However the increase (both absolute and percentage) in TWD was significantly larger in the PXF group. At 6 months, PXF produced a 292% increase in TWD (vs 180% produced by placebo) (p<0.02). The excess increase produced by PXF treatment was 112% at 6 months in comparison with placebo (p<0.02). Treatment was well tolerated. Between-group analysis favors PXF considering TWD, and results indicate good efficacy and tolerability. PMID- 11865836 TI - Treatment of venous ulcers with pentoxifylline: a 6-month randomized, double blind, placebo controlled trial. AB - The aim of this study was the evaluation of treatment with pentoxifylline in patients with venous ulcers in a 6-month, randomized, controlled trial. Treatment with placebo or pentoxifylline (PXF; 400 mg, 3 times daily) lasted 6 months and was associated to elastic bandaging. The endpoints were the number of limbs with complete healing and the variation in the area of ulceration. A group of 172 patients were included: 82 in the PXF group and 88 in the placebo group; 82 completed the study in the PXF group and 78 in the placebo group. Results. The two groups were comparable for age and sex distribution. The treatment was well tolerated. Complete healing was obtained in 67% of patients in the PXF group and 30.7% in the placebo group (p<0.02). The variations in the average area of ulceration were 86.7% (decrease) in the PXF group and 47% in the placebo group. The cost of treatment increased 21% with PXF but the cost due to non-healing of the ulcer was equivalent to a 44% increase (in comparison with the PXF group). In conclusion PXF is effective and cost-effective in improving ulcer healing in patients with chronic venous hypertension. PMID- 11865837 TI - Treatment of venous ulcers with pentoxifylline: a 12-month, double-blind, placebo controlled trial. Microcirculation and healing. AB - The aim of this study was the evaluation of the treatment of venous ulcers with pentoxifylline (PXF) in 12 months. This double-blind, placebo-controlled trial was conducted in GCP. Patients were included after consent and randomized for PXF (400 mg three times daily) or placebo. The number of healed ulcers, the variation in the area of ulceration, and the variation in microcirculatory parameters (laser-Doppler flux and skin PO2) were considered as endpoints. Of the 85 patients included, 80 completed the study (41 in the PXF and 39 in the placebo group). The groups were comparable. Healing occurred in 88% of limbs in the PXF and in 44% in the placebo group (44% difference; p<0.02). The average area reduction was 93% in the PXF and 56% in the placebo group. Microcirculation was significantly improved in both groups at 12 months. The decrease in flux and the increase in PO2 were greater in the PXF group (p<0.05). Considering the proportion of healed ulcers a better outcome was observed with PXF (p<0.02; 12% of nonhealed ulcers in the PXF vs 56% in the placebo group). The ulcerated area decreased 93% in the PXF vs 56% in the placebo group (40% difference; p<0.02). Tolerability was comparable (6% in the PXF group and 5% in the placebo group complained for treatment). No important side effects were observed. In conclusion, treatment with PXF in venous ulcers improves healing and microcirculation and should be considered in slow-healing venous ulcers. PMID- 11865838 TI - Treatment of intermittent claudication with pentoxifylline: a 12-month, randomized trial--walking distance and microcirculation. AB - The efficacy, safety and cost of pentoxifylline (PXF) in severe intermittent claudication was studied comparing PXF and placebo in a 12-month study. A treadmill test and microcirculatory evaluation with laser Doppler flowmetry were performed at inclusion and at the end of 6 and 12 months. A physical training plan (based on walking) and reduction in risk factor levels plan was used in both groups. Of the 120 included patients, 101 completed the study: 56 in the PXF group and 45 in the placebo group. There were 19 dropouts (due to low compliance). The two groups were comparable for age, sex distribution, walking distance, and the presence of risk factors and smoking. Intention-to-treat analysis indicated a 268% increase in walking distance in the PXF group (vs 198% in the placebo group; p<0.05) at 6 months and an increase of 404% (vs 280% in the placebo group; p<0.02) at 12 months. The absolute and percent increase in pain free walking distance (PFWD) was greater in the PXF group (p<0.05). Treatment was well tolerated. No serious drug-related side effects were observed. Microcirculatory evaluation indicated an increase in flux (p < 0.05) in the PXF group (not significant in the placebo group); the after-exercise flux (AEF) was increased (p<0.05) in both groups at 6 months but the increase in AEF was greater in the PXF group at 12 month. In conclusion, between-group analysis favors PXF considering walking distance and microcirculatory parameters. Results indicate good efficacy and tolerability. PMID- 11865839 TI - Ischemic renal disease in Kuwait. AB - Ischemic renal disease (IRD) is a frequent cause of end-stage renal disease. Its prevalence is mainly known from autopsy or retrospective arteriographic studies. This prospective study was conducted in 115 subjects selected from 732 patients with advanced chronic renal failure (CRF). Only patients with clinical features suggestive of IRD were selected for this study. In addition to detailed clinical and laboratory evaluation, captopril renal scintigraphy was performed in selected cases. All subjects underwent renal arteriography and all were followed up for 18.4 +/- 11.4 months. Renovascular disease was seen in 15 patients and significant bilateral renal artery disease leading to IRD was observed in 13 (11.3%). Hence the prevalence of IRD in the advanced CRF patients was 1.7%. The majority of patients with IRD (8 [61%]) were above 46 years of age and there were more men than women (10:3). Atherosclerotic renovascular disease was the most common (10 [77%]), even though arthritis (1 [7.6%]), and fibromuscular dysplasia (2 [15.3%]) were also observed. Serum creatinine at time of presentation was significantly higher in patients with IRD (784 +/- 292, p = 0.043) compared to those who did not have IRD (359 +/- 126). Corrective procedures were performed in 5 patients. After treatment the improvement in serum creatinine in patients with IRD at 3 and 6 months (166 +/- 32 and 173 +/- 47, respectively) was significantly different (p < or = 0.05) compared to those who were not treated (610 +/- 194 and 645 +/- 220, respectively). Hyperlipidemia, coronary artery disease and peripheral vascular disease were more prevalent in patients who had IRD compared to those with renal failure. The incidence of diabetes mellitus were similar in both groups. This study denotes a lower prevalence of IRD in the advanced CRF population; they had more severe renal failure at presentation but specific corrective treatment delayed progression of renal disease significantly. PMID- 11865841 TI - Abstracts of the 27th Annual Meeting of the American Society of Andrology. April 24-27, 2002. Seattle, Washington, USA. PMID- 11865840 TI - Emergency coronary stenting for complete thrombotic occlusion of an unprotected left main coronary artery in acute myocardial infarction complicated by cardiogenic shock in an octogenarian patient--a case report. AB - This report concerns an 82-year-old white man, who was admitted with cardiogenic shock secondary to an acute anterior myocardial infarction with right bundle branch block requiring an intra-aortic balloon pump for hemodynamic support and mechanical ventilatory support for respiratory distress. An immediate cardiac catheterization with coronary angiography revealed a complete thrombotic occlusion of the left main coronary artery. Prompt stent-supported percutaneous transluminal coronary angioplasty to the occluded left main coronary artery, a critical stenosis of the ostial left anterior descending artery, and the left circumflex coronary artery, allowed for recovery from this life-threatening condition and subsequent discharge from the hospital of this octogenarian patient. It is suggested that in a critical clinical condition with particularly challenging coronary anatomical findings, stent-supported coronary angioplasty can be lifesaving treatment in selected patients with octogenarian status with acute myocardial infarction. PMID- 11865842 TI - Expression of receptors for verotoxin 1 from Escherichia coli O157 on bovine intestinal epithelium. AB - Human enterohaemorrhagic Escherichia coli (EHEC) infection most commonly arises, either directly or indirectly, from cattle, which act as a reservoir host for these bacteria. In man, EHEC disease can be severe, whereas EHEC do not normally cause disease in cattle. Verotoxins (VTs) are the main virulence factors in human disease but no role for VT has been ascribed in cattle; however, this study shows for the first time that VT receptor is expressed by the bovine intestinal tract. VT bound to crypt epithelial cells of the small (ileum and jejunum) and large (caecum and colon) intestine independently of the animals' age. VT also bound to discrete cell subsets in the bovine kidney and to submucosal lymphoid cells but not to vasculature. Analysis of tissues for isoforms of the VT receptor, Gb3, confirmed the presence of the receptor in the bovine intestinal epithelium and kidney. A distinct pattern of Gb3 receptor isoform mixtures was observed in the bovine kidney. This, together with the general absence of receptors on vasculature, could contribute to the apparent resistance of cattle to systemic effects of VT. Expression of Gb3 on the bovine intestinal epithelium, together with previously described effects, may affect EHEC colonisation in its reservoir hosts and hence the potential for distribution to man. PMID- 11865843 TI - Evaluation of a simple and rapid dipstick assay for the diagnosis of typhoid fever in Indonesia. AB - To support the clinical diagnosis of typhoid fever in Indonesia, where most hospitals and health centres have no facilities for culture, a rapid dipstick assay for the detection of Salmonella typhi-specific IgM antibodies was evaluated on serum samples from 127 patients clinically suspected of having typhoid fever. In a single blood sample collected on admission to hospital, the sensitivity of the dipstick assay was 69.8% when compared with bone marrow culture and 86.5% when compared with blood culture. The specificity as calculated for the group of patients with suspected typhoid fever but a negative culture result was calculated to be 88.9%. Of 80 patients with febrile illnesses other than typhoid fever, reactivity was observed in only three patients with dengue haemorrhagic fever. The assay uses stabilised components that can be stored outside the refrigerator, does not require special equipment, and may be of use in remote health facilities that have no culture facilities. PMID- 11865844 TI - A world-wide internet survey of public knowledge about tuberculosis. AB - Four simple multiple-choice questions about tuberculosis (TB) were posted on a non-medical internet site for a 2-month period. A total of 564 responses were received. Sixty-two were excluded as individuals had made multiple attempts at the questions. Sixty-five per cent of responses were from North America, 14.5% from Europe and 12% from Australia and New Zealand, with only a small number of responses from Africa, the Indian subcontinent and South America. Of the respondents 49.5% correctly answered that cough is the commonest symptom of TB, 45% knew that TB was transmitted mainly by air-borne droplets, 37.8% knew that TB was caused by a bacterium. Only 19.5% knew that the most important risk factor for developing TB was HIV infection and only 4% answered all questions correctly. This survey suggests that knowledge about tuberculosis is limited in computer literate individuals throughout the world. PMID- 11865845 TI - Immunohistochemical staining for KIT (CD117) in soft tissue sarcomas is very limited in distribution. AB - We performed immunohistochemical analysis for KIT in 365 soft tissue sarcomas. Most tumors evaluated were completely negative for KIT, including all cases of leiomyosarcoma, rhabdomyosarcoma, myxofibrosarcoma, liposarcoma, solitary fibrous tumor, synovial sarcoma, dermatofibrosarcoma protuberans, schwannoma, malignant peripheral nerve sheath tumor, clear cell sarcoma, low-grade endometrial stromal sarcoma, and follicular dendritic cell sarcoma. Tumors showing occasional immunoreactivity for KIT included extraskeletal myxoid chondrosarcoma (2/20), Ewing sarcoma/malignant primitive peripheral neuroectodermal tumor (4/20), melanotic schwannoma (3/5), metastatic melanoma (4/20), and angiosarcoma (5/20). In most cases, staining for KIT was focal. Rare tumor cells showing KIT positivity were identified in a small number of other tumors. This study demonstrates very limited expression of KIT in soft tissue tumors other than gastrointestinal stromal tumors and underscores the discriminatory value of KIT immunohistochemical analysis for differential diagnosis. As some of these findings differ markedly from previous reports, it is evident again that variations in immunohistochemical technique can lead to major discrepancies in positive staining. Since treatment eligibility for selective tyrosine kinase inhibitors such as STI571 hinges on positive immunostaining, standardization and reproducibility of meaningful results are critically important. PMID- 11865846 TI - Nucleolated variant of mantle cell lymphoma with leukemic manifestations mimicking prolymphocytic leukemia. AB - Chronic lymphoproliferative disorders sometimes can be difficult to classify. We report 4 cases characterized by large cells with distinct central nucleoli, reminiscent of prolymphocytic leukemia, but shown on further workup to represent mantle cell lymphoma. At initial examination, the patients had generalized lymphadenopathy, splenomegaly, and a leukemic blood picture. The peripheral blood showed many large cells with round to slightly irregular nuclei, single central nucleoli, and a fair amount of pale cytoplasm. The picture was not typical of prolymphocytic leukemia because of the presence of generalized lymphadenopathy and the large size of the circulating abnormal cells. Immunophenotypic study showed that the large lymphoid cells were CD5+ CD23- mature B cells with overexpression of cyclin D1, and cytogenetic study demonstrated the translocation t(11;14)(q13;q32) in 3 patients. Lymph node biopsy confirmed a diagnosis of mantle cell lymphoma, pleomorphic variant, in all 4 patients. This study documents the existence of an unusual leukemic form of mantle cell lymphoma with prominent nucleoli; the clinicopathologic features that distinguish it from other chronic lymphoproliferative disorders are discussed. PMID- 11865847 TI - UK patients group hits back at anti-vivisectionists. PMID- 11865848 TI - Woman damaged by aspirin loses court claim. PMID- 11865849 TI - Harvard School of Public Health refuses tobacco funds. PMID- 11865850 TI - Urgent reform of NHS governance needed. PMID- 11865851 TI - Mandela presents his concerns about AIDS policy to ANC committee. African National Congress. PMID- 11865852 TI - United States starts to consider paying organ donors. PMID- 11865853 TI - Spain to ban drinking in the street. PMID- 11865855 TI - Forum calls for action on child health to tackle heart disease. PMID- 11865856 TI - Abstracts of the VIII World Congress on Endometriosis. San Diego, California, USA. February 24-27, 2002. PMID- 11865857 TI - Use of the Critical Thinking Disposition Inventory in critical care orientation. AB - In the current health care environment it is imperative that orientation programs be effective and efficient in the preparation of nurses entering critical care nursing. Institutions strive to develop orientation programs that use the least amount of resources necessary to achieve the desired outcome. The desired outcome of the critical care orientation process is for nurses to use critical thinking skills to make sound clinical judgments based on scientific knowledge of critical care nursing. Identifying areas of strengths and deficits in critical thinking could prove beneficial in assisting educators to individualize nursing orientation programs using critical thinking skills in practice. The purpose of this article is to describe a method used to measure nurses' dispositions toward critical thinking and the application of the California Critical Thinking Disposition Inventory (CCTDI) in critical care orientation programs. PMID- 11865858 TI - Postdiploma nursing education: after the year 2000. AB - The emergence of new health care needs related to changing demographics, financial restraints on health care, and increased rates of chronic illness has contributed to the need for nurses with a range of highly specialized skills. Yet traditional education programs for nurses have focused on diploma level preparation for bedside staff positions. This study explores the assessments of 714 clinical nurses with diploma level preparation and 56 nurse managers about the need for postdiploma nursing education. The results reveal a preference for clinically focused university education over noncredit coursework or a generic degree program. Nurses and their managers agree on the necessity for opportunities for more preparation in gerontology and palliative care, as well as the need for all practicing nurses to learn many skills cutting across clinical areas including clinical decision-making, communication, and leadership. Strategies for assisting nurses to access university education including innovative distance education methods are discussed. PMID- 11865859 TI - Implications of the situated learning model for teaching and learning nursing research. AB - Situated learning theory is gaining increased attention in the fields of human cognition and learning. The authors discuss the key researchers and basic assumptions of situated learning, and outline implications for the design and development of instruction. The authors describe how they applied the situated model to teaching and learning nursing research, an area identified as problematic for staff, administrators, and educators. They describe their personal observations and discuss feedback from participants at the workshop. The authors conclude that the situated model is useful for exposing nurses with little prior research experience to this domain of knowledge. PMID- 11865860 TI - Continuing education about physically abusive relationships: does education change the perceptions of health care practitioners? AB - BACKGROUND: The purpose of this study is to determine the effectiveness of mandatory continuing education for domestic violence. METHOD: This study surveyed health care professionals attending the 1993 and 1997 National Perinatal Association Conference in Florida to determine change their awareness and identification of pregnant women involved in physically abusive relationships. RESULTS: Even though in 1993 Florida initiated a mandatory 1-hour continuing education credit on domestic violence, the findings report that there were no statistically significant changes for awareness and identification. However, there was a statistically significant increase in the availability of patient education materials within the 4-year period. CONCLUSION: Both the 1993 and 1997 respondents expressed a desire for more professional education related to domestic violence. PMID- 11865861 TI - Health professionals' knowledge of women's health care. AB - Until 1986, the only issues in women's health which received direct attention were those related to childbearing. At that time the National Institutes of Health made inclusion of women in research a criteria for funding. Since then, the knowledge base to guide disease prevention and treatment of women has grown dramatically. Unfortunately, the incorporation of these data into clinical practice has been much slower. The purpose of this needs assessment was to establish a comprehensive data base on which future programming decisions could be based. Areas of interest identified by health care providers included stress reduction, breast health, prevention of heart disease, and osteoporosis. Other topics that emerged were menopause and reproduction issues, cancer prevention, domestic violence, substance abuse, nutrition, and weight control. These data suggest topics for future programming. PMID- 11865862 TI - Lessons learned: developing education for a system-wide documentation project. AB - BACKGROUND: As Summa Health System implemented an integrated interdisciplinary documentation system, educational sessions for staff had to be developed and presented. METHOD: Structure of classes, scheduling of sessions, ongoing revisions and refinements, and key elements of success are shared. RESULTS: Staff members incorporated the documentation changes into practice. CONCLUSION: Other organizations may benefit and learn from the experiences of our two-hospital system in developing and scheduling education for a system-wide change project. PMID- 11865863 TI - Abstracts from the 2001 Annual Conference of the Canadian Pain Society. May 10 12, 2001. Montreal, Quebec, Canada. PMID- 11865864 TI - Microbial contamination of spacecraft. AB - Spacecraft and space habitats supporting human exploration contain a diverse population of microorganisms. Microorganisms may threaten human habitation in many ways that directly or indirectly impact the health, safety, or performance of astronauts. The ability to produce and maintain spacecraft and space stations with environments suitable for human habitation has been established over 40 years of human space flight. An extensive database of environmental microbiological parameters has been provided for short-term (< 20 days) space flight by more than 100 missions aboard the Space Shuttle. The NASA Mir Program provided similar data for long-duration missions. Interestingly, the major bacterial and fungal species found in the Space Shuttle are similar to those encountered in the nearly 15-year-old Mir. Lessons learned from both the US and Russian space programs have been incorporated into the habitability plan for the International Space Station. The focus is on preventive measures developed for spacecraft, cargo, and crews. On-orbit regular housekeeping practices complete with visual inspections are essential, along with microbiological monitoring. Risks associated with extended stays on the Moon or a Mars exploration mission will be much greater than previous experiences because of additional unknown variables. The current knowledge base is insufficient for exploration missions, and research is essential to understand the effects of space flight on biological functions and population dynamics of microorganisms in spacecraft. Equally important is a better understanding of the immune response and of human microorganism-environment interactions during long-term space habitation. PMID- 11865865 TI - Plants, plant pathogens, and microgravity--a deadly trio. AB - Plants grown in spaceflight conditions are more susceptible to colonization by plant pathogens. The underlying causes for this enhanced susceptibility are not known. Possibly the formation of structural barriers and the activation of plant defense response components are impaired in spaceflight conditions. Either condition would result from altered gene expression of the plant. Because of the tools available, past studies focused on a few physiological responses or biochemical pathways. With recent advances in genomics research, new tools, including microarray technologies, are available to examine the global impact of growth in the spacecraft on the plant's gene expression profile. In ground-based studies, we have developed cDNA subtraction libraries of rice that are enriched for genes induced during pathogen infection and the defense response. Arrays of these genes are being used to dissect plant defense response pathways in a model system involving wild-type rice plants and lesion mimic mutants. The lesion mimic mutants are ideal experimental tools because they erratically develop defense response-like lesions in the absence of pathogens. The gene expression profiles from these ground-based studies will provide the molecular basis for understanding the biochemical and physiological impacts of spaceflight on plant growth, development and disease defense responses. This, in turn, will allow the development of strategies to manage plant disease for life in the space environment. PMID- 11865866 TI - Psychosocial issues in long-term space flight: overview. AB - Anecdotal evidence of the individual and interpersonal problems that occurred during the Shuttle-Mir Space Program (SMSP) and other long-duration Russian/Soviet missions, and studies of personnel in other isolated and confined extreme (ICE) environments suggest that psychosocial elements of behavior and performance are likely to have a significant impact on the outcome of long duration missions in space. This impact may range from individual decrements in performance, health and well being, to catastrophic mission failure. This paper reviews our current understanding of the psychosocial issues related to long duration space missions according to three different domains of behavior: the individual domain, the interpersonal domain and the organizational domain. Individual issues include: personality characteristics that predict successful performance, stress due to isolation and confinement and its effect on emotions and cognitive performance, adaptive and maladaptive coping styles and strategies, and requirements for the psychological support of astronauts and their families during the mission. Interpersonal issues include: impact of crew diversity and leadership styles on small group dynamics, adaptive and maladaptive features of ground-crew interactions, and processes of crew cohesion, tension and conflict. Organizational issues include: the influence of organizational culture and mission duration on individual and group performance, and managerial requirements for long duration missions. Improved screening and selection of astronaut candidates, leadership, coping and interpersonal skills training of personnel, and organizational change are key elements in the prevention of performance decrements on long-duration missions. PMID- 11865868 TI - Psychosocial issues in space: future challenges. AB - As the duration of space flights increases and crews become more heterogeneous, psychosocial factors are likely to play an increasingly important role in determining mission success. The operations of the International Space Station and planning of interplanetary missions represent important future challenges for how to select, train and monitor crews. So far, empirical evidence about psychological factors in space is based on simulations and personnel in analog environments (i.e. polar expeditions, submarines). It is apparent that attempts to transfer from these environments to space requires a thorough analysis of the human behavior specific to the fields. Recommendations for research include the effects of multi-nationality on crew interaction, development of tension within crews and between Mission Control, and prediction of critical phases in adaptation over time. Selection of interpersonally compatible crews, pre-mission team training and implementation of tools for self-monitoring of psychological parameters ensure that changes in mission requirements maximize crew performance. PMID- 11865867 TI - Psychosocial issues in space: results from Shuttle/Mir. AB - Important psychosocial issues involving tension, cohesion, leader support, and displacement of negative emotions were evaluated in a 4 1/2-year study involving five U.S. and four Russian Shuttle/Mir space missions. Weekly mood and group climate questionnaires were completed by five U.S. astronauts, eight Russian cosmonauts, and 42 U.S. and 16 Russian mission control subjects. There were few findings that supported our hypothesized changes in tension, cohesion, and leader support in crew and ground subjects using various time models, although crewmembers reported decreasing leader support in the 2nd half of the missions, and astronauts showed some evidence of a novelty effect in the first few weeks. There was no evidence suggesting a 3rd quarter effect among crewmembers on any of the 21 subscales evaluated. In contrast, there was strong evidence to support the hypothesized displacement of tension and negative emotions from crewmembers to mission control personnel and from mission control personnel to management. There were several significant differences in response between Americans vs. Russians, crewmembers vs. mission control personnel, and subjects in this study vs. people in comparable groups on Earth. Subject responses before, during, and after the missions were similar, and we did not find evidence for asthenia in space. Critical incidents that were reported generally dealt with events on-board the Mir and interpersonal conflicts, although most of the responses were from a relatively small number of subjects. Our findings have implications for future training and lead to a number of countermeasures. PMID- 11865869 TI - Clinostats and bioreactors. AB - The environment created on Earth within a clinostat or Rotating Wall Vessel (RWV) bioreactor is often referred to as "simulated microgravity". Both devices utilize constant reorientation to effectively nullify cumulative sedimentation of particles. Neither, however, can fully reproduce the concurrent lack of structural deformation, displacement of intercellular components and/or reduced mass transfer in the extracellular fluid that occur in actual weightlessness. Parameters including density, viscosity, and even container geometry must each be considered to determine the overall gravity-dependent effects produced by either a clinostat or the RWV bioreactor; in addition, the intended application of these two devices differs considerably. A state of particle "motionlessness" relative to the surrounding bulk fluid, which is nearly analogous to the extracellular environment encountered under weightless conditions, can theoretically be achieved through clinorotation. The RWV bioreactor, on the other hand, while similarly maintaining cells in suspension as they continually "fall" through the medium under 1 g conditions, can also purposefully induce a perfusion of nutrients to and waste from the culture. A clinostat, therefore, is typically used in an attempt to reproduce the quiescent, unstirred fluid conditions achievable on orbit; while the RWV bioreactor ideally creates a low shear, but necessarily mixed, fluid environment that is optimized for suspension culture and tissue growth. Other techniques for exploring altered inertial environments, such as freefall, neutral buoyancy and electromagnetic levitation, can also provide unique insight into how gravity affects biological systems. Ultimately, all underlying biophysical principles thought to give rise to gravity-dependent physiological responses must be identified and thoroughly examined in order to accurately interpret data from flight experiments or ground-based microgravity analogs. PMID- 11865870 TI - Human and rodent ground-based models of space flight environments. PMID- 11865871 TI - Consequences of contamination of the spacecraft environment: immunologic consequences. AB - Long-term space voyages pose numerous known and unknown health hazards, to the human immune system. Well-studied clinical examples of secondary immunodeficiencies created on Earth, lead one to predict that the conditions of prolonged space flight would weaken the human immune responses that normally hold infection and cancer in check. From evidence gathered from humans flown for prolonged periods in space and from human models of space flight studied on Earth it is reasonable to suspect that space travelers to the planet Mars would experience a weakening of immunity. Subtle defects of immune cell structure and function have been observed in astronauts, such as weakening of specific T lymphocyte recall of specific antigens. Ground-based models also have demonstrated alterations of immune function, such as the elevation of neuroendocrine immune system messengers, interleukin-6, and soluble tumor necrosis factor-alpha receptor in sleep deprivation. Since severe immune compromise the clinical consequences of reactivation of latent virus infections and the development of cancer, has yet to be seen in space flight or in the Earth models, it is extremely important to begin to quantify early changes in immunity to predict the development of immune system collapse with poor clinical outcomes. This approach is designed to validate a number of surrogate markers that will predict trouble ahead. Inherent in this research is the development of countermeasures to reduce the risks of infection and cancer in the first humans going to Mars. PMID- 11865872 TI - Transfer of multiple drug resistance plasmids between bacteria of diverse origins in natural microenvironments. AB - Plasmids harboring multiple antimicrobial-resistance determinants (R plasmids) were transferred in simulated natural microenvironments from various bacterial pathogens of human, animal, or fish origin to susceptible strains isolated from a different ecological niche. R plasmids in a strain of the human pathogen Vibrio cholerae O1 E1 Tor and a bovine Escherichia coli strain were conjugated to a susceptible strain of the fish pathogenic bacterium Aeromonas salmonicida subsp. salmonicida in marine water. Conjugations of R plasmids between a resistant bovine pathogenic E. coli strain and a susceptible E. coli strain of human origin were performed on a hand towel contaminated with milk from a cow with mastitis. A similar conjugation event between a resistant porcine pathogenic E. coli strain of human origin was studied in minced meat on a cutting board. Conjugation of R plasmids between a resistant strain of the fish pathogenic bacterium A. salmonicida subsp. salmonicida and a susceptible E. coli strain of human origin was performed in raw salmon on a cutting board. R plasmids in a strain of A. salmonicida subsp. salmonicida and a human pathogenic E. coli strain were conjugated to a susceptible porcine E. coli strain in porcine feces. Transfer of the different R plasmids was confirmed by plasmid profile analyses and determination of the resistance pattern of the transconjugants. The different R plasmids were transferred equally well under simulated natural conditions and under controlled laboratory conditions, with median conjugation frequencies ranging from 3 x 10(-6) to 8 x 10(-3). The present study demonstrates that conjugation and transfer of R plasmids is a phenomenon that belongs to the environment and can occur between bacterial strains of human, animal, and fish origins that are unrelated either evolutionarily or ecologically even in the absence of antibiotics. Consequently, the contamination of the environment with bacterial pathogens resistant to antimicrobial agents is a real threat not only as a source of disease but also as a source from which R plasmids can easily spread to other pathogens of diverse origins. PMID- 11865873 TI - A case for permitting altruistic surrogacy. AB - Canada's Royal Commission on New Reproductive Technologies rejects all forms of surrogacy arrangement under the rubric of objecting to commercial surrogacy. Noncommercial surrogacy arrangements, however, can be defended against the commission's objections. They can be viewed as cases of giving a benefit or service to another in a way that expresses benevolence, and establishes a relationship between surrogates and prospective 'social' parents that allows mutual understanding and reciprocal personal interaction between them. PMID- 11865874 TI - Sex-selective abortion: a relational approach. AB - A critical application of Ruddick's model of maternal thinking is the best way to grapple with the ethical dilemmas posed by sex-selective abortion which I view as a "moral mistake." Chief among these is the need to be sensitive to local cultural practices in countries where sex-selective abortion is prevalent, while simultaneously developing consistent international standards to deal with the dangers posed by the use of sex-selective abortion to eliminate female fetuses. PMID- 11865875 TI - Public and private eugenics: what can no longer be accomplished through overt government intervention may be largely achieved by private, indirect means. PMID- 11865876 TI - Eugenics, euthenics, euphenics: the underlying issue is that someone decides, based on values, which characteristics should be part of society and which are not. PMID- 11865877 TI - Preliminary report on informed consent for mental capacity assessments. AB - The goal of the study was to investigate the issue of informed consent for mental capacity. Seventeen clients referred to the Competency Clinic at the Baycrest Centre for Geriatric Care were interviewed after the assessment had been completed. Their responses to questions regarding the reasons for and possible consequences of the assessment indicated a range of understanding and of capacity to give informed consent. The results are discussed in terms of the appropriateness of having a fairly low threshold for informed consent in situations where other capacities are already in question. PMID- 11865878 TI - The ultimate test of autonomy: should minors have a right to make decisions regarding life-sustaining treatment? PMID- 11865879 TI - IVF and the internet. PMID- 11865880 TI - Disclosure preferences about terminal illness: an examination of decision-related factors. AB - Twenty-six male and 86 female, predominantly White, non-terminal cancer patients addressed preferences for disclosure of terminal prognosis, should their disease advance to that stage. Specific inquiries were made about desired levels of disclosure (full, partial,or non-disclosure) and desired pathways of disclosure (from physician to patient only, from physician to patient in the presence of a loved one, or from physician to loved one only). Gender, previous experience with death, and trait anxiety were associated with level preference. Education, previous experience with death, and trait anxiety were associated with pathway preference.Variables predictive of level and pathway preference were identified, benefits to physicians and patients were explored, and sampling limitations were discussed. PMID- 11865881 TI - Bereavement support for couples following death of a baby: program development and 14-year exit analysis. AB - Program development, implementation, and a 14-year exit analysis of a bereavement support program for couples whose baby died in the Neonatal Intensive Care Unit (NICU) is presented. A closed, hospital-based, time-limited (12 weeks) format was used. Team leadership was used and 54% of bereaved NICU parents participated. Each group was structured with a 2-week introductory period, open format grief focused weekly discussions,evaluation in Week 11, and summary session with termination in Week 12. The exit analysis details program strengths, weaknesses, and recommendations. Bereavement support groups are one part of what we contend should be a comprehensive bereavement program,organized to care for families prior to, during, and after a baby's death. A sensitive, spiritually aware, supportive environment should be maintained throughout with relationship building as a cornerstone of the program. PMID- 11865882 TI - Correlates of death anxiety in Pakistan. AB - To ascertain the effect of gender, age, and religiosity on death anxiety, 132 participants were interviewed using Templer Death Anxiety Scale and Collett Lester Fear of Death Scale (CLS). Women, older participants, and less religious participants were found to be more scared of their impending death. Gender effect was more pronounced, however, on the CLS. Women and less religious people reported to experience greater anxiety than their respective counterparts about different dimensions of death, for example, the shortness of life, total isolation of death, fear of not being, and disintegration of body after dying. The findings of the current work indicate that the general predictors of death anxiety, gender, age, and religiosity reported in Western, predominantly Christian samples also hold in an Eastern, Muslim sample. PMID- 11865883 TI - No-suicide contracts: an overview and recommendations. AB - No-suicide contracts, in their various forms, can deepen commitment to a positive action, strengthen the therapeutic alliance, facilitate communication, lower anxiety, aid assessment, and document precautions. Conversely, they can anger or inhibit the client, introduce coercion into therapy, be used disingenuously, and induce false security in the clinician. Research on no-suicide contracts (frequency surveys, assessments of behavior after contracting, and opinions of users) has limitations common to naturalistic studies, and is now ready for more rigorous methods. Mental health professions should be trained to deal with suicidal individuals, including how to use no-suicide contracts. Good contracts are specific, individualized, collaborative, positive, context-sensitive, and copied. However, they are not a thorough assessment, a guarantee against legal liability, nor a substitute for a caring, sensitive therapeutic interaction. No suicide contracts are no substitute for sound clinical judgment. PMID- 11865884 TI - 2.2 million Americans lost health insurance in 2001. PMID- 11865885 TI - New guidelines offer better diagnosis and treatment of ventilator-associated pneumonia. PMID- 11865886 TI - Evidence shows little benefit of treating middle ear infections with antibiotics. PMID- 11865887 TI - Primary stroke centers recommended; association develops implementation guidelines. PMID- 11865889 TI - Unlocking the mystery and misdiagnosis of central auditory processing disorder. PMID- 11865888 TI - Stent and glycoprotein inhibitor use linked to better outcomes and lower costs. PMID- 11865890 TI - American Heart Association revises guidelines for emergency care. PMID- 11865891 TI - Patients with low brain blood oxygen levels after surgery have longer stays. PMID- 11865892 TI - FDA issues final guidance on requirements governing single-use medical devices. PMID- 11865893 TI - Joint Commission releases new pain management standards. PMID- 11865894 TI - Audio weight loss program curbs obesity. PMID- 11865895 TI - Disease management program cuts costs, improves outcomes for Virginia Medicaid. PMID- 11865896 TI - Claims data may alert California plan to important treatment information. PMID- 11865897 TI - Guidelines introduced to help providers with newest HEDIS measure: menopause counseling. PMID- 11865898 TI - Updated guidelines include new drugs and therapies for the treatment of osteoarthritis. PMID- 11865899 TI - Lice treatment guidelines emphasize careful medication use. PMID- 11865900 TI - C-section guidelines offer benchmarks, easier rate standardization. PMID- 11865901 TI - Studies on pediatric length of stay guidelines draw controversy. PMID- 11865902 TI - Physicians more likely to face quality incentives than incentives that may restrain care. AB - Concerns that physician financial incentives may lead to withholding needed care have caught the attention of legislators, regulators and even the U.S. Supreme Court. While the spotlight has been on how health plans reimburse physician practices, this Issue Brief provides unique nationally representative data on physician practices' use of incentives, which have a more direct effect on physician behavior. According to 1999 data from the Center for Studying Health System Change (HSC), physicians are more likely to be subject to incentives that may encourage use of services, such as patient satisfaction (24 percent) and quality (19 percent), than to financial incentives that may restrain care, such as profiling (14 percent). The complexity of physician financial incentives and their relatively low prevalence raise questions about effective regulation and public reporting of their use. PMID- 11865903 TI - Consumers face higher costs as health plans seek to control drug spending. AB - Faced with relentless growth in pharmaceutical spending during the 1990s, health plans in recent years have tried to rein in costs by negotiating lower drug prices, encouraging more cost-conscious physician prescribing patterns and moderating the volume and mix of drugs demanded by consumers. Because of limited success with these strategies, plans have moved rapidly to three-tier benefit packages that offer broader drug choices but shift more costs to consumers. The move to three-tier pharmacy benefits appears to have slowed drug-spending growth for some plans--at least for the short term--but raises questions about the cost and quality of pharmaceutical care for consumers. Based on interviews with health plan executives in the 12 nationally representative communities the Center for Studying Health System Change (HSC) visits every two years, this Issue Brief examines plans' strategies to contain drug spending and the possible consequences for consumers. PMID- 11865904 TI - Employer health insurance premium subsidies unlikely to enhance coverage significantly. AB - State and local efforts to reduce the number of uninsured workers include three major approaches: public insurance expansions, subsidies paid directly to low income workers to help pay their share of employer-sponsored insurance premiums or buy individual insurance and subsidies paid directly to small employers to reduce the cost of health insurance premiums. Based on a national study by the Center for Studying Health System Change (HSC), premium subsidies paid directly to small firms are unlikely to significantly reduce the number of uninsured. About 16 million people work in firms with fewer than 50 workers that do not offer health insurance. A hypothetical 30 percent premium subsidy targeted to the employers of these workers--slightly more generous than the average in existing small firm subsidy programs across the country--would extend coverage to only about half a million uninsured workers if implemented nationally. PMID- 11865905 TI - Health care power of attorney and combined advance directive legislation (as of January 1, 1997). PMID- 11865906 TI - Physician-assisted suicide -- is it a constitutional right? PMID- 11865907 TI - Premium subsidies for employer-sponsored health coverage: an emerging state and local strategy to reach the uninsured. AB - With nearly 75 percent of the uninsured living in households with at least one full-time worker, there has been renewed policy interest in strategies to expand coverage by subsidizing employer-sponsored insurance. Six of the 12 nationally representative communities that the Center for Studying Health System Change (HSC) tracks have premium assistance, or subsidy, programs planned or underway. Policy makers are enthusiastic about the potential to expand coverage through these programs, but enrollment has been modest to date. This Issue Brief examines operational challenges facing subsidy programs, such as how to structure a benefits package within budgetary and regulatory constraints and how to attract employers and employees without displacing existing private contributions to premiums. It also discusses the trade-offs policy makers may face to resolve these challenges in the context of rising premiums and a slowing economy. PMID- 11865908 TI - Is managed care good for what ails you? Ruminations on race, age and class. PMID- 11865909 TI - Triple jeopardy: low income, chronically ill and uninsured in America. AB - At least 7.4 million working-age Americans with chronic conditions--such as diabetes, heart disease and depression--lacked health insurance in 1999, according to new research findings from the Center for Studying Health System Change(HSC). Uninsured people with chronic conditions report worse health and more functional limitations and are three times more likely not to get needed medical care compared to those who are privately insured. The vast majority of uninsured people with chronic conditions delayed or did not get needed care because of cost. About 63 percent of the uninsured with chronic conditions- roughly 4.7 million Americans--have family incomes below 200 percent of poverty, or about $35,000 a year for a family of four in 2001. Faced with the triple threat of low income, ongoing health problems and no health insurance, this group confronts great difficulty getting and paying for needed care. PMID- 11865910 TI - Options for expanding health insurance for people with chronic conditions. AB - As policy makers explore options such as tax credits and expansion of public programs to cover uninsured Americans, working-age adults with chronic conditions merit special attention. Because their medical needs are likely to be greater than healthy people's, coverage expansion proposals that don't factor in the greater need of people with chronic conditions are likely to fall short of reaching this vulnerable group. Often perceived primarily as a problem of the elderly, chronic conditions are widespread among working-age adults, according to a new study by the Center for Studying Health System Change (HSC). This Issue Brief examines existing coverage sources for insured working-age people with chronic conditions and assesses how various coverage proposals might affect uninsured people with chronic conditions. PMID- 11865911 TI - Empowering and protecting patients: lessons for physician-assisted suicide from the African-American experience. PMID- 11865912 TI - Birth and death: doctor control vs. patient choice. PMID- 11865913 TI - Pragmatism in the face of death: the role of facts in the assisted suicide debate. PMID- 11865914 TI - Symposium. Physician-assisted suicide: facing death after Glucksberg and Quill. Foreword: facing death. PMID- 11865915 TI - On the meaning and impact of the physician-assisted suicide cases. PMID- 11865916 TI - The death with dignity movement: protecting rights and expanding options after Glucksberg and Quill. PMID- 11865917 TI - Physician-assisted suicide in the courts: moral equivalence, double effect, and clinical practice. PMID- 11865918 TI - Disorder in the court: physician-assisted suicide and the Constitution. PMID- 11865920 TI - 'The freest nation in the world'? PMID- 11865919 TI - The future of euthanasia and physician-assisted suicide: beyond rights talk to informed public policy. PMID- 11865921 TI - Legal and policy issues in genetics and insurance. AB - Genetic screening and testing techniques provide a powerful diagnostic tool for the acquisition of predictive information. The potential value of such diagnostic techniques cannot be overstated. However, commercial organisations such as insurance companies and employers are also highly interested in the acquisition and use of genetic information. Concerns about the potential abuse of genetic information have stimulated a counter-current of public pressure for restrictions on the use which can be made of genetic diagnostic information. In a number of countries this pressure has generated enough concern to stimulate legislatures to enact laws which curtail the use and acquisition of genetic information. This pattern has clearly emerged in the United States of America and there are indications that similar trends are developing in Europe. This paper examines the law and policy issues arising from the interface between genetics and insurance. PMID- 11865922 TI - HUGO urges genetic benefit-sharing. AB - In view of the fact that for-profit enterprise exceeds public expenditures on genetic research and that benefits from the Human Genome Project may accrue only to rich people in rich nations, the HUGO Ethics Committee discussed the necessity of benefit-sharing. Discussions involved case examples ranging from single-gene to multi-factorial disorders and included the difficulties of defining community, especially when multifactorial diseases are involved. The Committee discussed arguments for benefit-sharing, including common heritage, the genome as a common resource, and three types of justice: compensatory, procedural, and distributive. The Committee also discussed the importance of community participation in defining benefit, agreed that companies involved in health have special obligations beyond paying taxes, and recommended they devote 1-3% of net profits to healthcare infrastructure or humanitarian efforts. PMID- 11865923 TI - The dangerous patient exception to the psychotherapist-patient privilege: the Tarasoff duty and the Jaffee footnote. AB - With the U.S. Supreme Court's 1996 decision in Jaffee v. Redmond, all U.S. jurisdictions have now adopted some form of evidentiary privilege for confidential statements by patients to psychotherapists for the purpose of seeking treatment. The majority of states, following the decision of the Supreme Court of California in Tarasoff v. Regents of the University of California, have also adopted some form of duty by psychotherapists to breach confidentiality and warn potential victims against foreseeable violence by their patients. Largely unresolved is whether there should be a dangerous patient exception to the evidentiary privilege parallel to the Tarasoff exception to confidentiality. This Article argues that exception to the evidentiary privilege should be evaluated separately from the exception to confidentiality. Whether or not a Tarasoff duty to warn existed at an earlier time, exception to the evidentiary privilege should be made only where psychotherapists' testimony is necessary to prevent future harm to patients or identified potential victims. Applying this standard, the dangerous patient exception generally would not apply in criminal actions against patients, but would apply only in proceedings for the purpose of protecting patients or third parties, such as restraining order hearings or proceedings to hospitalize patients. PMID- 11865924 TI - Permitting suicide of competent clients in counseling: legal and moral considerations. AB - State statutes, case law, and professional codes of ethics in the mental health professions typically stress either a duty or the permissibility of disclosing confidential information in order to prevent clients from seriously harming themselves. These sources are intended to address cases where clients are deemed to be suffering from cognitive dysfunction for which paternalistic intervention, including involuntary hospitalization, is considered necessary to prevent self destructive behavior. The counselor's moral and legal responsibility is less apparent when mentally competent clients desire suicide as release from irremediable suffering due to severe physical illness, and this desire is defensible within these clients' value systems. This paper will explore moral and legal dimensions of a counselor's decision not to intervene in such cases. The concept of permitted suicide will be introduced and defined, and guidelines for its application developed. PMID- 11865925 TI - British panel supports cloning of human cells for research. PMID- 11865926 TI - Changing practice based on evidence: changing education too. PMID- 11865927 TI - Developing interactive continuing education on the Web. AB - BACKGROUND: To meet the continuing education needs of advanced practice nurses, on-line continuing education modules were designed as part of a grant-funded nurse practitioner learning project. METHOD: Three modules were planned from the results of a nurse practitioner (n=187) survey of computer availability and skills, topic preference, and their potential for taking on-line continuing education. RESULTS: The development and implementation of two of the three modules demonstrates the interactive framework, extensive use of links, pre- and posttesting, and on-line registration. The two modules were designed to be interactive, realistic self-studies that closely resemble clinical practice. CONCLUSIONS: Outcome data are currently being collected on-line and will be analyzed for efficacy of this delivery system for continuing education. PMID- 11865928 TI - The impact of AIDS continuing education on psychiatric and non-psychiatric nurses' knowledge. AB - BACKGROUND: Because an increasing number of chronically mentally ill individuals are HIV-positive, continuing education must include AIDS content to ensure psychiatric nurses provide competent care. The Fundamentals of Mental Health and HIV/AIDS Program was tested to determine its effectiveness in augmenting the AIDS knowledge of mental health professionals. METHOD: Two hundred forty RNs attended the Fundamental of Mental Health and HIV/AIDS Program. They completed pretest and posttest measures. RESULTS: The program significantly increased AIDS knowledge. The increase in AIDS knowledge was more dramatic for psychiatric nurses than for non-psychiatric nurses. CONCLUSION: This study reinforces the need for AIDS continuing education. PMID- 11865929 TI - Learning group: facilitating the adaptation of new nurses to the specialty unit. AB - BACKGROUND: The nursing shortage creates major problems in finding qualified, experienced nurses in the critical care area. METHOD: A learning group was formed to expedite the new nurses' acquisition of skills and knowledge needed to care for critical care patients. RESULTS: After 4 months of continuing education provided in the learning group, all group members had demonstrated their interest in learning. They also had demonstrated they were able to perform different procedures and skills according to the standards listed in the trauma intensive care unit nurses' manual. CONCLUSION: The learning group is able to facilitate quicker adaptation and smoother transition of new nurses to the trauma intensive care unit. The continuing education provided by the learning group demonstrated support from the workplace and peers to the new nurses. PMID- 11865930 TI - Polished, professional presentation: unlocking the design elements. AB - Using available computer graphics software programs, a novice can develop a presentation. Going beyond the basics, however, requires knowledge of the use of the elements of design: fonts, colors, and graphics. This article describes how to select fonts and colors and provides guidelines for use of pictures and graphs. Guidelines for use of default settings and templates, and considerations related to the use of animation, sound, and projection are included. Examples are provided to demonstrate important points and creating a polished, professional product. PMID- 11865931 TI - Continuing education in nursing: a 10-year retrospective study of CE offerings presented by the Kentucky Nurses Association. AB - BACKGROUND: The purpose of this retrospective study was to evaluate continuing education (CE) offerings presented by the Kentucky Nurses Association during a 10 year period. METHOD: Data were collected and analyzed from 37 CE offerings (N = 474 respondents). RESULTS: Findings revealed that the majority of the respondents reported that the CE offerings were useful, stimulated them to seek further information, and resulted in acquisition of significant knowledge or skills for nursing education or practice. However, fewer than half of the respondents reported they were able to implement changes in nursing education or practice as a result of the CE offerings. CONCLUSION: Findings supported the usefulness of CE in nursing education and practice and the need for further research evaluating CE and the correlation between CE and nursing education and practice. PMID- 11865933 TI - Faculty practice in joint appointments: implications for nursing staff development. AB - Faculty joint appointments can capitalize on the talents and unique knowledge of nurses in varied settings. Altered role expectations, recurring emphases on cost control, and demands for accountability in the transforming health care system urge academic nursing faculty and clinically-based nurses to work together to achieve requisite health care and educational outcomes. This article addresses the types of faculty practice models, discusses three major role types of joint appointments, and presents the benefits and challenges of faculty functioning in joint appointments. Implications for nursing staff development are examined. PMID- 11865932 TI - Australian nurse educators identify gaps in expert practice. AB - In Australia, nurses face a double-barreled challenge to their role. With the rapid adoption of new health care technologies coupled with increasing economic constraints, they find themselves "doing more with less." In this context of continuous change, it is useful to determine what expert nurse clinicians deem the most essential skills, attitudes and knowledge required for practice in complex technological environments. Separate panels of 28 educators and 43 cardiac nurse clinicians participated in a national Delphi study rating the importance to the nursing role of 107 items drawn from the international literature on expert practice and technology. Indicating the importance of each item in both the "real" and "ideal" worlds of practice, educators identified 58 items where they felt actual practice was substantially far from the ideal. For 16 of these items relating to empowerment of patients, nursing research, and technology policy, the educators rated clinical behavior below the median of the real world scale, indicating substandard performance of a role or inadequate assimilation of a concept. The implications for the definition of expert practice and for curricula development are discussed. PMID- 11865934 TI - Conditions of anesthesia practice on the Ivory Coast. AB - The Ivory Coast is one of the most developed countries in Africa, but conditions of practice of anesthesia are difficult. From anesthesia equipment to medications that are available, all resources are limited, and nurse anesthetists as well as sole anesthesia providers, have to manage with isolation and lack of information in uncomfortable situations. PMID- 11865935 TI - Starting a nurse anesthesia program in Palestine. AB - In spite of the fact that nonphysician anesthesia providers are used in many areas of the world, some countries continue to have shortages in the anesthesia workforce. As a result of many requests from around the world for information on starting a nurse anesthesia program, the Education Committee of the International Federation of Nurse Anesthetists (IFNA) developed a document entitled "Guidelines for Starting A New Program and Sample Curriculum." It was hoped that this document, along with IFNA's "Educational Standards for Preparing Nurse Anesthetists," would be of assistance to individuals starting programs internationally. This article is a case example of necessary steps in establishing a nurse anesthesia program. In addition to beginning stages and organization of the school, it describes the challenges encountered. It highlights the importance of follow-up and continued assistance for success of new programs. PMID- 11865936 TI - Volunteer activities in developing countries: Haiti, Costa Rica, Uganda, Eritrea, and Guyana. AB - I have volunteered 16 times in 5 different developing countries. Each of these trips have involved my profession as a CRNA. My students have been health care providers of different types from these countries. Fourteen of these trips have been in conjunction with Health Volunteers Overseas. I have taught clinical and didactic anesthesia and helped develop nurse anesthesia programs in Uganda and Guyana. The purpose of this activity has been to increase the quality of health care in these countries by helping to provide safer anesthesia. PMID- 11865937 TI - The volunteer anesthetist: a personal view. AB - The most common opportunities for nurse anesthetists to be involved in volunteer anesthesia overseas are usually on a surgical team to provide direct care for selected types of patients or to be involved in teaching local anesthesia providers. The challenges are numerous and unique in each setting. Sharing of knowledge, directly as an instructor or indirectly as a provider, provides for a great impact on health care delivery in many emerging nations. The anesthesia provider who is working to deliver care in another country must consider many variables before taking on such a venture. The type of surgical team or teaching assignment one accepts will determine the type of preparation that must be done before departing. In addition to the many organizational items, one should consider some personal issues as well. Lost wages, transportation costs, and lodging expenses are often paid for by the volunteer. Time away from one's family may also be a consideration. Health care, such as updated immunizations, must be attended to before to departure. Volunteers must be able to provide good care in less than ideal situations, and often with much less equipment and medications than are available in the United States. A review of some of the issues that one should consider before deciding to be a volunteer are outlined in this article. The nurse anesthetist who successfully anticipates and deals with the challenges of volunteer anesthesia will be rewarded with an intense degree of personal satisfaction. The ability to give back to the profession and those who would otherwise not have access to one's skills or knowledge produces a sense of accomplishment that is unique. PMID- 11865938 TI - The International Federation of Nurse Anesthetists: its role in the globalization of nurse anesthesia education and practice. AB - Since its beginning in 1989, the International Federation of Nurse Anesthetists (IFNA) has had as its major goal in the advancement of educational standards and practices that support and enhance patient care and safety. This article highlights the development of the IFNA, its purpose, and its structure. Educational and practice standards are discussed and quality assurance in international education highlighted. PMID- 11865939 TI - Untoward pathophysiological events: simulation as an experiential learning option to prepare anesthesia providers. AB - Untoward pathophysiological conditions coupled with anesthesia delivery can lead to critical incidents. Anesthetists must be armed with a set of complex skills to respond effectively in acute situations. A survey of nurse anesthetists was conducted in which respondents were asked to report the frequency that they had experienced one of 24 select events in their practice in the last 60 days. They were asked to rank how meaningful it would be for graduate students to experience the same or a similar event in their training in order to be prepared for practice. Also, they were asked to rate, among 6 proposed curricular changes, those which would best address student learning needs. Seven events were identified as having occurred more than 13 times; hypertension, hypotension, bradycardia, acute hemorrhage, hypothermia, coronary constriction, and oliguria. The mean score for how important it was thought for students to experience untoward events in their training was 4.11, with a range of 3.54 for Addisonian crisis to 4.54 for experience in managing hypertension. Recommended curricular change scores ranged from a low of 3.33 for adding more theory in course work to 4.58 for adding simulator experience in the laboratory for critical events. This article identifies anesthetist learning needs from the practitioner's perspective and assists programs in adapting curricula accordingly. Overwhelmingly, respondents believe experiential learning is required to prepare anesthetists to manage untoward events. The rare occurrence of most critical events is a strong rationale for the use of simulator technology as the preferred method to address these learning needs. PMID- 11865940 TI - Anesthetic drug interactions: quarterly update. PMID- 11865942 TI - Nurse anesthesia practice and research--a worldwide need. AB - In 1992, nurse anesthetists from 94 countries participated in our transnational research by completing surveys (in English, French, German, or Spanish) and providing descriptions about their practice, education, and regulation. Five years later (1997), nurse anesthetists from 81 countries completed expanded and revised questionnaires. Their responses validated the 1992 findings that nurses may be the main administerers of anesthesia in many countries. They are administering both general and regional anesthesia, in rural and urban areas in all regions of the world, and in countries from all levels of development, working with and without anesthesiologists. Their major duties are primarily those that are performed inside operating rooms, a factor that may be contributing to a lack of visibility of the profession. In order to contribute to both clinical and health policy decision making, nurse anesthetists must first document their practice and then participate in collaborative research at the local, regional, national, and international levels. PMID- 11865941 TI - Thoughts on the "new nursing shortage". AB - The nursing shortage is a frequently recurring phenomena in the United States and in many developed countries worldwide. It is time that serious consideration be given to the necessary changes that are essential for supplying an adequate nursing force worldwide. This article compares the late 1980s nursing shortage in the United States with the current one and finds that many of the same causes are prevalent. These causes stem from the gender-based history of modern nursing and the cultural- and professional-based problems that nursing has had great difficulty changing and are discussed in this article. The decentralization of health care has also created greater demand for nurses. The nursing component of a hospital's budget representing its largest component has also been targeted by hospital administrations and ownership as a source for supporting other departments or functions at the expense of nursing personnel. Finally, the vast opportunities that have opened up to women since the Civil Rights Act in the 1960s and today's booming economy and high-technology environments make nursing much less attractive and competitive in today's market. Also, the managed care movement has significantly contributed to problems of nursing. Nursing educators have also found that the 3 levels of nursing education has confused today's high school graduates, leading to their rejection of nursing as a career. Thus, efforts are underway in nursing education to correct this problem--one that is beginning to be corrected through role differentiation that is now seen in many hospitals, but most prevalent in so-called "magnet" hospitals. Investigators have done much research--first in distinguishing the difference between magnet and nonmagnet hospitals, and now, regarding patient outcomes from those hospitals. These can lead to the efforts of nursing to come to grips with multiple entry points for a single licensure. However, the problems associated with the barriers to practice, prompted by long outdated mores and cultural influences discussed in this article that must change if nursing is to remain a viable and attractive profession in the 21st century. Investigators have found higher mortality rates between magnet and nonmagnet hospitals, favoring the magnet hospitals that have a better nurse-to-patient ratio, higher baccalaureate registered nurse staffs, and greater nursing autonomy. It is time that decision makers take heed and make changes. Nothing less than the lives of patients are at stake. PMID- 11865943 TI - Health care and regional anesthesia education for nurse anesthetists in Ghana. AB - In this article, an overview of economical, educational, and health care conditions in Ghana are discussed. The foundation for an educational course in regional anesthesia is laid, steps are taken for development and implementation is identified. Benefits of the experience to the people of Ghana are identified and personal rewards highlighted. PMID- 11865944 TI - Analysis of genetic aberrations in uterine adenomyosis using comparative genomic hybridization. AB - OBJECTIVE: To use comparative genomic hybridization (CGH) to analyzefrozen tissuesfrom adenomyosis cases to discover and map genomic regions for chromosomal gains and losses. STUDY DESIGN: In a retrospective study, upper and lower threshold values of 1.20 and 0.80, respectively, were used to define positive findings. RESULTS: No positive recurrent gene copy number alterations were detected in the 25 cases of pathologically proven adenomyosis. CONCLUSION: Although CGH is extremely useful in investigating candidate genes in the development of adenomyosis, CGH was not useful in this study. Genetic changes might be indeed extremely rare in adenomyosis, or CGH was not sensitive enough to detect candidate genes. PMID- 11865945 TI - Measuring microvascular density in tumors by digital dissection. AB - OBJECTIVE: To develop and validate a digital dissection techniquefor measuring the cross-sectional area of blood vessels in histologic sections of tumors routinely stained with hematoxylin and eosin. STUDY DESIGN: The procedure was first validated in four experimental tumors in rats by comparing the results of the digital dissection technique to functional estimates of the blood volume in the tumors as measured by dynamic, contrast-enhanced magnetic resonance imaging. The method was then tested on a variety of experimental and human tumors. RESULTS: The digital dissection technique yielded results that exactly matched the functional measurements of blood volume infour experimental tumors. Digital dissection of 40 additional tumors in rats showed that 21 infiltrating ductal carcinomas had significantly greater microvascular density (MVD) than 19 benign fibroadenomas (12% vs. 7.9%, P=.028 by two-tailed t test). In 10 human breast carcinomas the MVD was consistently greater than the measurement of blood vessel density as identified by immunohistochemical staining for factor VIII. The between-run coefficients of variation for the MVD assay were 12% (n = 5) for a human breast cancer and 18% (n = 5)for an experimental rat tumor. CONCLUSION: The digital dissection technique is a reproducible, objective and accurate method of measuring MVD in sections of tumors that are routinely stained with hematoxylin and eosin. PMID- 11865946 TI - DNA cytometric analysis of surgically treated squamous cell cancer of the uterine cervix, stage pT1b1-pT2b. AB - OBJECTIVE: To determine the utility of DNA content and DNA-related variables of proliferative activity regarding prognosis in cervical cancer. STUDY DESIGN: DNA image (ICM) andflow cytometry (FCM) were performed to determine the DNA index (DI), 5c-exceeding rate (5c-ER), S-phase fraction (SPF) and proliferation index (PI) in 163 patients with surgically staged pT1b1-pT2b squamous cell cancer of the uterine cervix and treated with primary radical hysterectomy. ICM was performed on imprint cytology, obtained from fresh tumor tissue, which was also used for FCM. Results were analyzed using the chi2 test and Cox regression analysis for risk of pelvic lymph node involvement, tumor recurrence and recurrence-free survival (RFS). RESULTS: ICM was performed on all 163 and FCM on 133 samples. One-third of the tumors showed DNA aneuploidy. Analysis demonstrated prognostic significance of a DI > or = 1.70, with a (70:30) 2.3-fold risk of recurrence (P=.024) and reduced RFS of 10 months (P=.003) in cases of DI > or = 1.70. A high 5c-ER > 11% was associated with pelvic lymph node involvement and decreased RFS (P < or = .04). Significantly more relapses were found in tumors with SPF > 12% (70.8% vs. 29.2%, P=.007). RFS was markedly reduced in tumors with high SPF (52.3 vs. 61.1 months, P=.011). Low proliferative tumors (PI<25%) were associated with lower stage (P=.036) and increased RFS (61.2 vs. 47.1 months, P=.028). In multivariate analysis of clinicopathologic variables (pT category, nodal status, lymphovascular space involvement) and DNA related variables, pelvic lymph node involvement was the only significant predictor of RFS. In patients with nodal involvement, tumors with DI >1.70 were associated with lessfavorable outcomes. CONCLUSION: DNA-related variables of cell cycle analysis were valuablefor predicting prognosis in cervical cancer patients. Tumors with DI>1.70, 5c-ER >11% and high proliferative activity (SPF>12%, PI>25%) represent a subgroup with a poor prognosis. PMID- 11865947 TI - Potential of the learning vector quantizer in the cell classification of endometrial lesions in postmenopausal women. AB - OBJECTIVE: To investigate the potential of artificial neural networks for cell identification in endometrial lesions from postmenopausal women. STUDY DESIGN: The study was performed on cytologic material obtained by the Gynoscann endometrial cell samplerfrom 12 cases of atrophic endometrium, 48 cases of hyperplasia without cytologic atypia (18 cases of simple hyperplasia and 30 cases of complex hyperplasia), 12 cases of hyperplasia with cytologic atypia (complex atypical hyperplasia) and 48 cases of adenocarcinoma (30 cases of well differentiated, 12 cases of moderately differentiated and 6 cases of poorly differentiated carcinoma). From each case approximately 100 cells were examined using a custom image analysis system. A learning vector quantizer (LVQ) identified the collected data. RESULTS: Investigation of cells from Endometrial Alterations with LVQ proved that according to the nuclear characteristics, as expressed by morphometric and textural measures, the endometrial cells from postmenopausal women may be identified as belonging to one of thefollowing three groups: atrophy, hyperplasia without cytologic atypia (simple and complex hyperplasia) and malignant neoplastic lesions (atypical complex and adenocarcinoma). CONCLUSION: The role of nuclear morphologic features in the cytologic diagnosis of endometrial alterations was confirmed. The overlap in thefeature space observed indicates that cell characteristics do not form strictly separate clusters. Thatfact explains the difficulty that morphologists have with the reproducible identification of cells from endometrial lesions in postmenopausal women. Application of LVQ offers a good classification at the cell level and promises to be a powerful toolfor classification on the individual patient level andfor the clarification of the natural history of endometrial pathology. PMID- 11865949 TI - Prognostic evaluation of breast cancer in cytologic specimens. AB - OBJECTIVE: To evaluate prognostic factors in breast cancer using cytologic samples and to determine the correlation between those factors and ploidy. STUDY DESIGN: Two hundred sixteen fine needle aspirates from patients with primary breast cancer were analyzed for expression of estrogen receptors (ERs), progesterone receptors (PRs), Ki-67 antigen, expression of p53 tumor suppressor gene and overexpression of c-erbB-2 using a standard immunochemical method. Not all subjects had all biomarker information because of the study design (c-erbB2 added later). The specimens were analyzed also for ploidy. We used the SAMBA 4000 image analysis system for quantification of the percent of cells stained positively by the different immunocytochemical stains andfor ploidy. RESULTS: A significant correlation wasfound between ER and PR and between Ki-67 and positive p53. Steroid receptor content was not significantly related to p53, Ki-67 or c erbB2. No correlation was found between c-erbB2 and the other biomarkers. Ploidy had a significant correlation with all the biomarkers used. CONCLUSION: A reliable and rapid evaluation of markers for breast cancer can be achieved by measuring cells stained positively by immunocytochemical stains, as well as ploidy, by means of an image analysis system. ER, PR Ki-67, p53 and c-erbB2 had a significant correlation with ploidy and overall prognostic value in breast cancer. PMID- 11865948 TI - Quantification of vascular density using a semiautomated technique for immunostained specimens. AB - OBJECTIVE: To develop a semiautomated, quantitative techniquefor the assessment of vascular density in immunohistochemically stained tissue sections using diaminobenzidine tetrahydrochloride (DAB) and hematoxylin as chromagens. STUDY DESIGN: A semiautomated thresholding technique was developed to quantitate vascular density in tissue sections stained with anti-CD31 (1 degrees antibody). The immunohistochemically stained specimens were digitally imaged using a 24-bit color camera. The blue component of the RGB image was segmented using a variable high-pass filter. After thresholding, the segmented areas (CD31 positive) were quantified and vascular density determined. The validity of the method was verified by calculating the precision of the technique using the coefficient of repeatability and by quantifying its agreement with manual analysis according to the Bland-Altman approach. RESULTS: Vascular endothelial cells were specifically selected using anti-CD31 as the primary antibody and the appropriate horseradish peroxidase-conjugated secondary antibody. Utilizing the semiautomated thresholding technique, the separation of DAB-stained tissuefrom non-DAB-stained tissue was achieved. The method developed possesses a low coefficient of repeatability (0.49%), agrees well with manual assessment (mean difference = 0.29 +/- 0.92%), is highly automated and is user friendly. CONCLUSION: A novel semiautomated technique for the quantification of vascular density was developed. This technique provides a method for reproducible measurement of immunostaining procedures (immunohistochemistry, immunocytochemistry and in situ hybridization) utilizing immunoperoxidase techniques with DAB as a chromagen. PMID- 11865950 TI - Immunohistochemical evaluation of HER-2/neu expression in infiltrating breast carcinoma: a study of reproducibility. AB - OBJECTIVE: To determine interobserver and intraobserver reproducibility in the assessment of the HercepTest- and TAB250-immunostained slides. STUDY DESIGN: Three independent expert pathologists (two with and one without training in HercepTest assessment) evaluated the HercepTest and TAB250-immunostained slides of 108 infiltrating breast carcinomas with a triple-blind method. The evaluation was repeated, with the same method and sequence of view, after 60 days. RESULTS: Expert pathologists, after adequate training in HercepTest evaluation, could reach excellent interobserver (K=.911, P<.001) and intraobserver reproducibility (K of .863-.926; P <.001 for all). The percentage of disagreement in intraobserver reproducibility ranged from 0.9% to 3.7%. Interobserver and intraobserver reproducibility in the evaluation of TAB250-immunostained slides was good (K = .658, P < .001) and from good to excellent (K of .600-.895, P < .001 for all), respectively. CONCLUSION: Optimization of the level of accuracy in HercepTest evaluation is mandatory because the decision to initiate therapy with Herceptin depends on the result. Moreover, considering that the percentage of disagreement in intraobserver reproducibility ranges from 0.9% to 3.7%, it is advisable that two expert pathologists evaluate all HercepTest slides with a double-blind method. If there are discordant results, they must be discussed by the same pathologists. PMID- 11865951 TI - Comparison of quantitative histomorphometry and DNA ploidy in tissue sections of prostate carcinoma. AB - OBJECTIVE: To examine the ability of quantitative histomorphometry to predict DNA ploidy of prostate carcinoma in biopsy tissue sections assigned after quantitation by nuclear digital image analysis. STUDY DESIGN: Thirty-five diploid, 35 tetraploid and 35 aneuploid prostatic carcinomas in biopsies, assessed by the CAS 200 image analyzer (Bacus Laboratories, Lombard, Illinois, U.S.A.), were reevaluated by the Bacus Laboratories Incorporated Slide Scanner, a microscope that quantifies histologic images. Thirty-one histomorphometric features from cancer cells were captured at 40 x magnification, averaged across tilesfor each case and incorporated into a multivariate discriminant model to determine which features predicted ploidy interpretation by nuclear image analysis using the CAS 200. RESULTS: On average, 60 and 15 minutes were required to perform nuclear image analysis and histomorphometry, respectively. The multivariate discriminant model identified configurable run length, difference variance, contrast, inverse difference moment, sum entropy and diagonal variance as histomorphometry features capable of distinguishing diploid from nondiploid tumors (P < .05). Cross-validation studies showed the model correctly classified 74.3% of the diploid and 57.1% of the nondiploid cases. CONCLUSION: Quantitative histomorphometry can predict the ploidy of prostate carcinoma in biopsy tissue sections. Quantitative histomorphometry has potential as a method of rapidly assessing DNA ploidy otherwise earmarked for nuclear image analysis, resulting in savings of time and expense. PMID- 11865952 TI - The special senses. PMID- 11865953 TI - Squamous cell carcinoma of the conjunctiva: clinicopathological features in 287 cases. AB - BACKGROUND: Squamous cell carcinoma is the most frequently encountered malignant tumour of the conjunctiva. The objective of this study was to describe the clinicopathological features of patients with squamous cell carcinoma of the conjunctiva seen at a large ophthalmologic hospital in Mexico City. METHODS: We reviewed the clinical and pathological files of all patients with documented squamous cell carcinoma of the conjunctiva seen at the hospital between 1957 and 1996. RESULTS: A total of 287 cases (286 patients) were reviewed. The mean age of the patients was 60.4 (range 12 to 99) years; 55% were male. The clinical diagnosis was accurate in 41% of cases. Typical keratinized squamous cell carcinoma of the conjunctiva accounted for 98% of the lesions, and there were small numbers of histologic variants: lymphoepithelioma-like carcinoma (three cases), spindle cell carcinoma (two cases) and mucoepidermoid carcinoma (one case). Evidence of local extension of the tumour was found in 150 patients (52%), with the cornea being most frequently involved (108 cases [38%]). Regional metastasis was found in two patients, to a submandibular lymph node in one and to a preauricular lymph node in the other. The most common form of treatment was local resection (258 cases [90%]). The mean length of follow-up was 7.7 (range 2 to 24) months. The recurrence rate was 5.2%. INTERPRETATION: We report a large series of patients with squamous cell carcinoma of the conjunctiva. The incidence of local extension was high. Three cases of lymphoepithelioma-like carcinoma, a variant not previously reported in the conjunctiva, were encountered and were confirmed by immunohistochemical reactions. PMID- 11865954 TI - Allergic reactions to brimonidine in patients treated for glaucoma. AB - BACKGROUND: Allergic reactions to ophthalmic drugs have not been studied extensively in ophthalmology. We performed a study to estimate the incidence of allergy to brimonidine in patients treated for glaucoma. METHODS: We identified all patients in a private glaucoma practice who started therapy with brimonidine between Mar. 19, 1998, and Aug. 14, 1999. We recorded the patient's diagnosis, age, sex, concomitant glaucoma medication, previous allergy to glaucoma medication and allergy to brimonidine. Allergy was defined as allergic contact dermatoconjunctivitis or follicular conjunctivitis. RESULTS: Of the 140 patients identified, 36 (25.7%) had had an allergic reaction to brimonidine. Contact dermatoconjunctivitis was noted in 19 patients (52.8%) and follicular conjunctivitis in 18 (50.0%). The rate of development of those two manifestations was linear and almost parallel throughout the study period. In logistic regression analysis previous allergy to a topically given antiglaucoma medication (t = -5.13) and concurrent use of levobunolol (t = 3.46) were retained as the most probable predictor variables of allergy to brimonidine. Life-table analysis showed a fairly linear curve, with no peak in allergy rate. Allergic reactions occurred throughout the year, with a small peak in March. INTERPRETATION: We found a rate of allergy to brimonidine of 25.7%. Concomitant levobunolol use and allergy to another glaucoma medication were associated with a higher allergy rate. PMID- 11865955 TI - Ultrasound biomicroscopy in eyes with anterior chamber flattening after trabeculectomy. AB - BACKGROUND: Information about changes in the trabeculectomy ostium and adjacent structures related to flattening of the anterior chamber in the early postoperative period may be important for management. We assessed the ostium and adjacent structures by means of ultrasound biomicroscopy (UBM) in order to identify common patterns related to postoperative flattening of the anterior chamber. METHODS: We studied 15 eyes of 15 consecutive patients. A first UBM examination was performed when the patient was referred. Changes in the ostium (evidence of tissue adhesions to the ostium) were evaluated with UBM after the chamber flattening resolved, at least 60 days after the first examination. The following features were noted: depth of the anterior chamber, size and patency of the ostium, rotation of ciliary processes, presence of uveal effusion and ultrasound functioning of the bleb. RESULTS: All the patients had ring-shaped ciliochoroidal effusions; in nine eyes the effusions were found only with UBM. In four eyes an anterior-posterior deformation of the ostium was found. Synechiae in the ostium were identified in the three eyes in which iris remnants had been recognized at the site on the initial UBM examination. INTERPRETATION: Our observations confirm the presumed pathophysiology of flat anterior chamber without bleb leak. Ring-shaped effusions were detected on UBM even in cases in which conventional ultrasonography showed no positive results. UBM is a helpful tool in the diagnosis and management of flattening of the anterior chamber after trabeculectomy. PMID- 11865956 TI - Persistent visual loss following retrobulbar hemorrhage. PMID- 11865957 TI - Wernicke's encephalopathy in hyperemesis gravidarum. PMID- 11865958 TI - Multiple epidermal inclusion cysts of the eyelid: a simple technique for removal. PMID- 11865959 TI - The spectrum of postmortem ocular findings in victims of shaken baby syndrome. PMID- 11865960 TI - Hydroxyapatite and calcium phosphate coatings on aluminium oxide orbital implants. AB - BACKGROUND: Hydroxyapatite and calcium phosphate have been used as bone graft substitutes as they facilitate and promote tissue ingrowth. We carried out a study to examine uncoated and coated aluminium oxide (alumina) spherical orbital implants and assess whether the coatings influence fibrovascular ingrowth. METHODS: The aluminium oxide spheres (three coated with hydroxyapatite, three coated with calcium metaphosphate and three uncoated) were manufactured at the School of Materials Engineering, Yeungnam University, Kyongsan, Kyongbuk, Korea. The implants were examined macroscopically and with scanning electron microscopy and were analysed chemically by means of x-ray powder diffraction and x-ray fluorescence spectrophotometry. Implantation of three hydroxyapatite-coated, three calcium metaphosphate-coated and three uncoated aluminium oxide spheres was done in nine adult male New Zealand albino rabbits. Implant vascularization was evaluated at 4, 8 and 12 weeks by means of histopathological sectioning. RESULTS: All three types of implant had multiple interconnected pores. The coatings increased the size of the trabeculae from 150 microm to 300 microm. As a result, the pores appeared slightly smaller but still ranged in size from 300 microm to 750 microm, compared to 400 microm to 800 microm in the uncoated implants. The coatings also increased the weight of the implants slightly. The implants were all strong mechanically. They were made up primarily of aluminium oxide. The coated implants contained significant amounts of calcium oxide (a contaminant). There was no clinical difference in the socket response between the three groups. Histopathologically, fibrovascularization occurred uniformly throughout each implant at 4, 8 and 12 weeks after implantation. INTERPRETATION: The hydroxyapatite and calcium metaphosphate coatings did not appear to facilitate or inhibit fibrovascular ingrowth at 4, 8 and 12 weeks. Longer-term studies are need to determine whether the coatings play a role in long-term acceptance and retention of the implants. PMID- 11865961 TI - The influence of hormones and pharmaceutical agents on DHEA and DHEA-S concentrations: a review of clinical studies. AB - Low endogenous levels of dehydroepiandrosterone (DHEA) and/or its sulfoconjugated derivative DHEA-S have been associated with diseases such as lupus, cancer, and diabetes. Circulating concentrations of DHEA and DHEA-S resulting from endogenous production or hormone supplementation may also be relevant in psychiatric illness. Drugs may significantly increase or decrease circulating concentrations of these adrenal androgens by various mechanisms. Some agents, such as dexamethasone, affect the HPA axis by inhibiting ACTH and therefore decrease DHEA and DHEA-S concentrations. Central nervous system agents, including carbamazepine and phenytoin, induce the P450 enzymes that metabolize DHEA and DHEA-S and therefore decrease circulating concentrations of these hormones. Danazol alters the ratio between DHEA and DHEA-S by inhibiting sulfatase. As research moves forward to better understand the relationships of these adrenal androgens with health and disease, it is essential that studies be designed to control for the influence of administered pharmaceuticals on DHEA and DHEA-S. PMID- 11865963 TI - Pemoline ingestion in children: a report of five cases and review of the literature. AB - The authors describe five pediatric cases of excessive pemoline ingestion. Based on their experience compared with previously reported cases in the literature, they describe the clinical presentation and rational treatment recommendations for acute pemoline ingestion. Overall, patients experienced a relatively benign clinical course following pemoline ingestion. Symptoms of pemoline ingestion appear to be primarily an accentuation of the drug's pharmacological effects on the central nervous and cardiovascular systems with sinus tachycardia, hypertension, hyperactivity, choreoathetoid movements, and hallucinations being most commonly observed. These findings are consistent with previously reported cases. Possible rhabdomyolysis manifested by evaluation of serum CPK was also observed in 3 of 4 patients in whom this laboratory parameter was measured and appears to be a common finding in acute pemoline poisoning. After acute ingestion, symptoms occurred within 6 hours, lasting up to 48 hours in all patients. Gastric lavage and/or activated charcoal would be effective decontamination measures, whereas ipecac-induced emesis should be avoided after massive ingestion due to the possibility of seizures. Aggressive use of a benzodiazepine appears a reasonable first choice to treat associated involuntary movements, tremor, hyperactivity, irritability, and agitation. Phenothiazines or butyrophenones may also be used especially for serious life-threatening symptoms, including hypertensive crisis and severe hyperthermia, although these serious complications of stimulant overdose have not been reported after pemoline ingestion. If a patient should experience pemoline-induced hypertensive crisis, individual dose titration of labetalol or sodium nitroprusside would appear reasonable pharmacologic approaches for rapid stabilization of blood pressure. PMID- 11865962 TI - Serum concentrations, efficacy, and safety of a new, intravenously administered varicella zoster immune globulin in pregnant women. AB - Chickenpox is teratogenic in humans, and varicella zoster immune globulin (VZIG) is given to pregnant women believed to be susceptible to the virus after contact with chickenpox. Available VZIG is given as intramuscular injections. The objective of this study was to evaluate the efficacy, safety, and serum concentrations of a new VZIG that can be given intravenously. The new VZIG (Cangene Pharm., Inc.) was compared to the standard VZIG (Massachusetts Public Health Biologic Laboratories) in a randomized protocol in 57 pregnant women seronegative to varicella zoster virus (VZV). Pregnant women received 125 units per 10 kg body weight to a maximal dose of 625 units. Women were evaluated on days 2, 7, 14, and 28 and at other times if symptoms developed into clinical varicella, which was scored by the Constitutional Illness Score. The new VZIG was comparable to the standard VZIG on all parameters of efficacy and safety. Levels of VZV antibodies at day 2 postinjection were significantly higher among those receiving the new preparation intravenously. The authors concluded that the new intravenous form of VZIG confers higher initial levels of VZV antibodies and is comparable in terms of its maternal efficacy and safety to the standard form of VZIG. PMID- 11865964 TI - Influence of food intake and formulation on the pharmacokinetics and metabolism of bosentan, a dual endothelin receptor antagonist. AB - The purpose of the study was to investigate the effect of food intake on the pharmacokinetics and metabolism as well as the relative bioavailability of bosentan. Sixteen healthy male subjects were treated in a randomized, four-way, crossover design with single oral doses of 125 mg bosentan, given as one tablet (with or without food), two tablets of 62.5 mg (with food), and a suspension (without food). The pharmacokinetic parameters of bosentan (and also three of its metabolites) were very similar after the four treatments: geometric means for Cmax and AUC0-infinity, ranged from 1.3 to 1.6 microg/ml and from 7.8 to 8.9 microg x h/ml, respectively, and median t(max) from 3.0 to 4.0 hours. The bioavailability of the 125 mg tablet relative to that of the suspension, both given fasted, was 102%. In the presence of food, Cmax and AUC0-max increased by 22% and 10%, respectively, whereas the two 62.5 mg tablets were bioequivalent to the 125mg tablet, both under fed conditions. The pharmacokinetics of the metabolites was independent of the treatment administered. In conclusion, bosentan bioavailability from the newly developed 125 mg tablet formulation is similar to that of the suspension, and food intake does not influence its pharmacokinetics to a clinically relevant extent. PMID- 11865965 TI - Short-term effects of new synthetic conjugated estrogens on biochemical markers of bone turnover. AB - Fifty early postmenopausal women completed a double-blind, placebo-controlled study evaluating the short-term effect of a new synthetic conjugated estrogens formulation (Cenestin) on bone turnover. Subjects were randomized to either 0.625 mg/day synthetic conjugated estrogens (n = 35) or placebo (n = 15) for 3 months. Biochemical markers were evaluated at baseline (three measurements at Days -2, 1, and 0) and Days 30, 60, and 90. Bone resorption assessed by urinary NTX ( 31.4%) and serum CTX (-34.2%) was significantly (p < 0.01) decreased in the estrogen-treated group compared to the placebo group within 1 month of treatment. The mean percent decreases for urinary NTX from baseline during estrogen treatment were -58.0% (p < 0.01 vs. placebo) and -34.1% (ns) after 2 and 3 months, respectively. For serum CTX, the percent changes from baseline were 17.6% (p < 0.01) and -16.9% (p < 0.01) at 2 and 3 months, respectively. As expected, the decrease of both bone formation markers (bone ALP and PINP) was delayed compared to that of bone resorption and significant (p < 0.05-0.01) only after 2 months of treatment in the estrogen-treated group compared to the placebo group. Synthetic conjugated estrogens significantly decreased bone resorption and bone formation comparable to that previously reported for estrogen treatments proven efficacious in preventing postmenopausal bone loss. PMID- 11865966 TI - The new phosphodiesterase 4 inhibitor roflumilast is efficacious in exercise induced asthma and leads to suppression of LPS-stimulated TNF-alpha ex vivo. AB - Roflumilast is a new phosphodiesterase 4 (PDE4) inhibitor developed by Byk Gulden Pharmaceuticals for the treatment of chronic obstructive pulmonary disease and asthma. A placebo-controlled, randomized, double-blind, two-period crossover study was performed to investigate the safety and efficacy of roflumilast in 16 patients with exercise-induced asthma. The patients received placebo or roflumilast (500 microg/day) for 28 days, each according to the randomly determined treatment sequences roflumilast/placebo and placebo/roflumilast. In both study periods, exercise challenge was performed 1 hour after dosing on days 1, 14, and 28. FEV1 was measured before exercise challenge, immediately after the end of exercise challenge, and then at 1, 3, 5, 7, 9, and 12 minutes after the end of challenge. Blood samples for the determination of lipopolysaccharide (LPS) stimulated tumor necrosis factor alpha (TNF-alpha) in whole blood ex vivo as a surrogate marker for the inhibition of inflammatory cell activation were taken predose on days 1 and 28. Serial safety measurements were performed during both study periods. Analysis of variance for the crossover design showed a significant superiority of roflumilast over placebo on day 28. The mean percentage fall of FEV1 after exercise was reduced by 41% as compared to placebo (p = 0.021). An improvement of lung function during roflumilast treatment was also observed on days 1 and 14. The median TNF-alpha level decreased by 21% (p = 0.009) during roflumilast treatment but remained essentially constant under placebo. It is concluded that roflumilast is effective in the treatment of exercise-induced asthma. This result was accompanied by a significant reduction of TNF-alpha levels ex vivo. Treatment with roflumilast was safe and well tolerated. PMID- 11865967 TI - Effect of calcineurin inhibitor therapy on P-gp expression and function in lymphocytes of renal transplant patients: a preliminary evaluation. AB - Cyclosporine and tacrolimus are substrates and potent inhibitors of the multidrug transporter, P-glycoprotein, in vitro. The authors have investigated the effect of chronic therapy with these and other immunosuppressive drugs on the expression and function of P-glycoprotein in T lymphocytes. Using a P-gp antibody, the authors studied the level of expression of P-gp in CD4 and CD8 T cells over a period of time in renal transplant patients. For comparison, a group of healthy volunteers and patients who did not receive any calcineurin inhibitors but were maintained on mycophenolate mofetil was included. The P-gp expression on lymphocytes from these two groups remained constant (over several months' time). However, patients who were started on tacrolimus or cyclosporine had an initial decline in expression of P-gp on CD4 T cells. Patients who were initiated on calcineurin therapy on day 1 posttransplant also had a decrease in expression of P-gp on CD4 T lymphocytes. This preliminary analysis suggests that the calcineurin inhibitors might be modulating the expression and function of transporters in lymphocytes, thus changing not only the drug concentration but also the apparent efficacy of these drugs. Further understanding and elucidation of such effects would be important in understanding the relationship between pharmacokinetics and pharmacodynamics of these and other drugs, especially for immunosuppressive and anti-AIDS therapy. PMID- 11865968 TI - Pharmacokinetics of fluvastatin in subjects with renal impairment and nephrotic syndrome. AB - The pharmacokinetics (PK) and safety of fluvastatin, a hydroxymethylglutaryl coenzyme A reductase inhibitor, were assessed in subjects with renal impairment and nephrotic syndrome. In a single-center, open-label, parallel-group study, a single dose of fluvastatin 40 mg was administered to subjects (8 per group, n = 48) with nephrotic syndrome (group II), healthy subjects (group I), and subjects with various degrees of renal impairment (groups III to VI). Subjects undergoing hemodialysis received two doses, one 2 days before and one just prior to hemodialysis. Blood samples to determine the PK parameters of fluvastatin were collected from 0 to 12 hours after drug intake. Noncompartmental PK evaluation and statistical analysis (descriptive and ANOVA) were performed. Safety was evaluated and vital signs were monitored. There was no difference in the PK parameters AUC0-infinity and Cmax of fluvastatin between healthy subjects and subjects with renal impairment. Fluvastatin was not removed from plasma by hemodialysis. In patients with nephrotic syndrome, the values for AUC0-infinity and Cmax were less than half of those obtained in the other groups; terminal half life values, however, were comparable. Fluvastatin was well tolerated in all study participants. Only few adverse events of mild to moderate intensity were reported. There were no clinically relevant changes in laboratory parameters in the subjects with renal impairment. Renal impairment did not affect the PK of fluvastatin after a single oral dose. Exposure to fluvastatin was lower in subjects with nephrotic syndrome. Fluvastatin also was well tolerated in subjects with nephrotic syndrome. PMID- 11865969 TI - A comparison of methods for assessing hypothalamic-pituitary-adrenal (HPA) axis activity in asthma patients treated with inhaled corticosteroids. AB - Suppression of the hypothalamic-pituitary-adrenal (HPA) axis is an accepted indicator of potential side effects from inhaled corticosteroids. Although cortisol monitoring is frequently used to detect changes in HPA axis activity, the optimal method for identifying the subset of asthma patients on inhaled steroids who experience severe cortisol suppression of potential clinical significance has not been established. The objective of this study was to compare several methods for assessing HPA axis activity in asthma patients taking inhaled corticosteroids. After screening, 153 patients with mild to moderate asthma were randomly assigned to receive inhaled fluticasone propionate (110, 220, 330, or 440 microg bid), flunisolide (500 microg or 1000 microg bid), or one of two control regimens (prednisone or placebo) for 21 days. Salivary (8 a.m.) and urinary (24-h) cortisol determinations were compared against 22-hour area under the serum cortisol concentration-time curve (AUC0-22 h) measured at baseline and on day 21. Comparisons were also made against 8 a.m. serum cortisol. A significant positive correlation was found between AUC0-22 h of serum cortisol and 8 a.m. serum cortisol level (r = 0.5140; p = 0.0001). The AUC0-22 h of serum cortisol was weakly correlated with 24-hour urinary cortisol levels, both corrected (r = 0.4388; p = 0.0001) and uncorrected (r = 0.3511; p = 0.0001) for creatinine excretion. The 8 a.m. salivary cortisol level correlated positively with the 8 a.m. serum cortisol level (r = 0.5460; p = 0.0001). Salivary cortisol was both sensitive and specific for the detection of a 50% decline in AUC0-22 h of serum cortisol. Cortisol reductions of this magnitude have been observed following repeated use of inhaled steroids. Because it is noninvasive, salivary cortisol measurement offers distinct advantages as a screening method for detecting pronounced HPA axis suppression in asthma patients receiving corticosteroid therapy. PMID- 11865970 TI - Pharmacokinetics of diethylcarbamazine after single oral dose at two different times of day in human subjects. AB - In most Wuchereria bancrofti and Brugia malayi infections, the microfilaria are found in the blood in greatest number between 10 p.m. and 2 a.m., indicating that chronotherapy may be beneficial in treating such infections. This study reports the influence of time of administration on the pharmacokinetics of diethylcarbamazine (DEC) in healthy volunteers. The study was conducted in 12 healthy volunteers by administering a 150 mg single oral dose of diethylcarbamazine citrate at 0600 or 1800 h in a balanced crossover design with the approval of an institutional ethics committee. The subjects fasted for about 10 hours before and 3 hours after drug treatment. Blood samples were collected at predetermined time intervals, and the drug content in the serum was estimated using HPLC with an electrochemical detector. Pharmacokinetic analysis was performed using noncompartmental methods employing WinNonlin (version 3.1), and the means of various pharmacokinetic parameters were compared for any dosing time related changes using a paired t-test at a probability level of 95%. The mean +/- SD values of pharmacokinetic parameters of DEC for the treatments at 0600 versus 1800 h were as follows: Cmax, 500+/-227 versus 637+/-401 ng/ml; tmax, 2.3+/-0.7 versus 2.7+/-1 h; Ka, 2.23+/-0.72 versus 1.96+/-0.97 h(-1); t1/2, 14.6+/-6.7 versus 11.4+/-4.9 h; AUC0-t, 5,334+/-1,853 versus 6,901+/-4,203 ng x h/ml; AUC0 infinity, 5,840+/-1,922 versus 7,220+/-4,205 ng x h/ml; CL/F, 36,058+/-19,011 versus 32,189+/-25,293 ml/h/kg; Vd/F, 570+/-225 versus 533+/-447 L; and MRT 17.7+/-5.9 versus 15.3+/-5.2 h. None of the parameters was significantly changed (p > 0.05) as a function of time of administration. PMID- 11865971 TI - Evaluation of single- and multiple-dose pharmacokinetics of synthetic conjugated estrogens, A (Cenestin) tablets: a slow-release estrogen replacement product. AB - A multiple-dose, placebo-controlled, randomized pharmacokinetic study was performed in 15 early (i.e., 1-3 years) postmenopausal women to evaluate the single and steady-state pharmacokinetics of 0.625 mg Cenestin (Synthetic Conjugated Estrogens, A) tablets, administered once daily for 90 days. Plasma concentration-time profiles for both total (conjugated and unconjugated) estrone and equilin, two major estrogens in Cenestin, were nearly superimposable between Day 1 (single dose) and Day 90 (multiple dose), indicating equivalent drug exposure from one dose to the next. For total estrone, the mean estimates of Cmax and AUC0-24 were 2.5 ng/ml and 35.0 ng x h/ml for Day 1 and 3.0 ng/ml and 39.8 ng x h/ml for Day 90, respectively. Similarly, Cmax and AUC0-24 mean values for total equilin were 1.4 ng/ml and 17.4 ng x h/ml after Day 1 and 1.5 ng/ml and 17.3 ng x h/ml after Day 90, respectively. The mean tmax value for total estrone was 8.3 hours on Day 1 and 8.6 hours on Day 90, indicating a slower rate of absorption. The average estimate for observed drug accumulation index for the 24 hour dosing interval was calculated to be 1.1 for total estrone and 1.0 for total equilin. These data, taken together, indicate a slow and steady rate of absorption, minimal drug accumulation at steady state, and consistent drug exposure between Cenestin doses. PMID- 11865972 TI - Lack of interaction between amiodarone and mexiletine in cardiac arrhythmia patients. AB - Amiodarone has pharmacokinetic interactions with various therapeutic agents, including phenytoin, flecainide, and cyclosporine. Mexiletine is metabolized by CYP2D6 and CYP1A2. The objective of this study is to evaluate the effect of amiodarone on the pharmacokinetics of mexiletine through its inhibition of various cytochrome P450 (CYP) subtypes. In a series of 181 inpatients with supraventricular tachyarrhythmias, 26 inpatients received mexiletine and amiodarone therapy (MEX + AMD group), and the others received mexiletine therapy (MEX group). In 10 inpatients of the MEX + AMD group, the mexiletine clearance (CL(MEX)/F) before and after coadministration of amiodarone was compared. CL(MEX)/F was also compared in the MEX and MEX + AMD groups after the start of amiodarone therapy. Serum mexiletine, amiodarone, and desethylamiodarone concentrations were measured by an HPLC method. The CL(MEX)/F was estimated by the Bayesian method using population pharmacokinetic analysis. There was no significant difference in CL(MEX)/F before and after 1-month coadministration of amiodarone in 10 inpatients of the MEX + AMD group. Although serum amiodarone and desethylamiodarone concentrations gradually increased with time after the start of amiodarone therapy in these patients, CL(MEX)/F showed no change at 3 and 5 months after the start of amiodarone therapy. There was no significant difference in CL(MEX)/F of the MEX group and the MEX + AMD group. The results suggest that the pharmacokinetics of mexiletine is not affected by amiodarone in patients with cardiac arrhythmias. PMID- 11865973 TI - 100 years of peroxynitrite chemistry and 11 years of peroxynitrite biochemistry. AB - The paper on the unusual properties of a mixture of hydrogen peroxide and nitrous acid by Baeyer and Villiger from 1901 can be regarded as the first report on peroxynitrite. In 1990, Beckman and co-workers suggested that peroxynitrite, formed from the reaction of superoxide with nitrogen monoxide, could be a transient oxidizing species in vivo, a report that revolutionized investigations in the field of oxidative stress. PMID- 11865974 TI - Hypochlorous acid-modified low-density lipoprotein inactivates the lysosomal protease cathepsin B: protection by ascorbic and lipoic acids. AB - Unregulated uptake of oxidized LDL by the scavenger receptor(s) of macrophages is thought to be an early event in atherosclerotic lesion development. Accumulation of oxidized LDL within macrophages may result from resistance of the modified LDL to enzymatic hydrolysis or from direct inactivation of lysosomal enzymes by reactive LDL-associated moieties. Since HOCl-modified LDL has been detected in vivo, the effects of HOCI-modified LDL on the activities of the cysteine protease cathepsin B and the aspartyl protease cathepsin D were investigated. LDL (0.5 mg protein/ml), which had been exposed to HOCl (25-200 microM), caused rapid dose dependent inactivation of cathepsin B, but not of cathepsin D. Exposure of LDL to HOCl results primarily in the formation of LDL-associated chloramines, and the model chloramine N(alpha)-acetyl-lysine chloramine also caused dose-dependent inactivation of cathepsin B. Incubation of HOCl-modified LDL with ascorbic and lipoic acids (25-200 microM) resulted in dose-dependent reduction of LDL associated chloramines and concomitant protection against cathepsin B inactivation. Thus, the data indicate that HOCl-modified LDL inactivates cathepsin B by a chloramine-dependent mechanism, most likely via oxidation of the enzyme's critical cysteine residue. Furthermore, small molecule antioxidants, such as ascorbic and lipoic acids, may be able to inhibit this potentially pro atherogenic process by scavenging LDL-associated chloramines. PMID- 11865976 TI - Development of a fecal occult blood test using a monoclonal antibody to haptoglobin. AB - Monoclonal antibodies (mAbs) were produced to human haptoglobin by immunising with fecal extracts from patients with colon cancer. An enzyme-linked immunosorbent assay was developed with one of the mAbs (FE14.1), and its ability to diagnose colorectal carcinoma evaluated. Patients with colorectal cancer were positive (43/46 = 93.5%) compared to normal individuals (4/211 = 1.9%). The assay has a specificity 93.5% and sensitivity 98.1% and has several advantages over current fecal occult blood tests. The test is potentially useful for bowel cancer diagnosis and to quantitate the level of haptoglobin in other body fluids such as urine and in effusions. PMID- 11865977 TI - Haptoglobin, inflammation, and tumorigenesis in the MIN mouse. AB - We have determined that the MIN mouse, which is predisposed to adenomas of the small and large intestine, exhibits chronic inflammation characterized by the induction of haptoglobin (HP), as well as other acute phase proteins. Inflammation is initiated at about 40-70 days of age, and is maintained throughout the life of the animal. Delayed onset of inflammation brought on by dietary means is associated with a reduction in tumor number and longer life span. Knockout mice that lack haptoglobin exhibit an increase in tumor number, indicating that the acute phase reactant suppresses tumorigenesis, perhaps through inhibition of the inflammatory response. The MIN mouse is, therefore, a useful model for studying the roles of HP and inflammation in tumorigenesis in vivo. PMID- 11865975 TI - BCL-2 is involved in preventing oxidant-induced cell death and in decreasing oxygen radical production. AB - It has been hypothesized that programmed cell death is mediated, in part, through the formation of free radicals via oxidative pathways. Furthermore, it has been proposed that BCL-2 acts to inhibit cell death by interfering with the production of oxygen-derived free radicals induced by a wide variety of stimuli. In order to examine the antioxidant function of BCL-2, we transfected mouse epidermal cells JB6 clone 41 with the expression vector pD5-Neo-BCL-2 and studied the effect of BCL-2 overexpression on oxidant-induced cell death and on the production of reactive oxygen species. Compared to Neo control cells, BCL-2-expressing cells are more resistant to the killing and growth retardation induced by hydrogen peroxide, superoxide, or by the oxygen radical-generating quinone-containing compounds menadione, diaziquone and adriamycin. The latter compounds generate reactive oxygen species during bioreductive metabolism. In addition, the exposed cells die by necrosis rather than apoptosis. Hydroxyl radical levels generated by the quinone-containing agents were low in BCL-2-expressing JB6 cells compared to control Neo cells. BCL-2, however, does not change the activities of the major cellular antioxidant enzymes superoxide dismutase, catalase or glutathione peroxidase. On the other hand, the glutathione concentrations increased in BCL-2 overexpressing cells after oxidative challenge, while the opposite was true for control cells. Thus, our results suggest that BCL-2 inhibition of oxidant-induced cell death is mediated, at least in part, through an antioxidant pathway, and that this pathway involves glutathione. PMID- 11865978 TI - Haptoglobin and the Th1/Th2 balance: hints from in vitro and in vivo studies. AB - A variety of immunomodulatory effects have previously been attributed to haptoglobin (Hp). These are supposed to be partly mediated through binding of Hp to CD11b. In the present study, we assessed its effects on T-helper (Th) cytokine production following both in vitro and in vivo stimulation of T-cells. Hp exhibits a dose-dependent inhibitory effect on human T lymphocyte release of the Th2 cytokines (IL-4, IL-5, IL-10 and IL-13) in vitro, whereas it has no clear effect on Th1 cytokine (IL-2 and IFN-gamma) release. When administered an anti CD3 monoclonal antibody, Hp knockout mice produced more IL-4 and less IFN-gamma than did their wild-type litter-mates. Our findings imply that Hp may be regarded as a regulator of the Th1/Th2 balance in both human and murine immune systems. PMID- 11865979 TI - Haptoglobin in lung defence. AB - Haptoglobin (Hp) has been known to be associated with the host defence response to infection and inflammation. The biological functions of Hp can be related to its ability to bind haemoglobin or to modulate immune response. Hp is expressed at a high level in lung cells, yet its protective role(s) in the lung is not known. Using transgenic mice overexpressing Hp, we demonstrated that Hp can reduce blood-induced lung injury. Hp-mediated haemoglobin catabolism in lung cells appears to be linked to iron mobilization, and may be an efficient mechanism to reduce oxidative damage associated with haemolysis. PMID- 11865980 TI - Consequences of haemolysis without haptoglobin. AB - Haptoglobin (Hp), a conserved plasma glycoprotein, forms very stable soluble complexes with free plasma haemoglobin. Haemoglobin binding by haptoglobin is thought to be important in the rapid hepatic clearance of haemoglobin from the plasma and in the inhibition of glomerular filtration of haemoglobin. It is thought to reduce haemoglobin-induced renal damage during haemolysis. To evaluate these functions, Hp knockout (Hp-/-) mice were created. The Hp-/- mouse was generated by a standard gene replacement technique in mouse embryonic stem cells. These mice were evaluated with and without haemolysis using several parameters: mortality, haemoglobin clearance, renal tissue damage and function. Hp-/- mice were viable but had a small, significant reduction in postnatal viability. The lack of Hp did not impair clearance of free plasma haemoglobin. Induction of severe haemolysis by phenylhydrazine caused extensive haemoglobin precipitation in the renal tubular cells. However, haemoglobin precipitation in the kidney was not increased in Hp-/- mice. Nevertheless, Hp-/- mice were more susceptible to phenylhydrazine with a mortality rate of 55% in Hp-/- mice versus 18% in Hp+/+ mice. In general, phenylhydrazine-treated Hp-/- mice suffered greater tissue damage, as evidenced by the induction of a hepatic acute phase response, resulting in increased plasma alpha1-acidic glycoprotein (AGP) levels and higher plasma malonaldehyde (MDA) and 4-hydroxy-2(E)-nonenal (HNE) levels. Gross pathological analysis indicated that the kidney was the most affected tissue in phenylhydrazine-treated Hp-/- and Hp+/+ mice, and Hp-/- mice were more severely affected. They had lower mitotic indices in their kidneys, higher basal levels of renal lipid peroxidation, as evidenced by levels of malonaldehyde and 4-hydroxy 2(E)-nonenal (MDA/HNE) and elevated levels of 8-hydroxyguanine (but not other products of oxidative DNA damage). There also was increased induction of haem oxygenase-1. The more severe renal damage in Hp-/- mice was also evident in the delayed erythropoietin gene expression and poorer renal clearance of [3H]-inulin. The reduction in glomerular filtration function in Hp+/+ and Hp-/- mice could be restored to baseline by vasodilators (prazosin or diazoxide), implicating renal vasoconstriction as a major mechanism of acute renal failure during induced haemolysis. These data suggest that Hp plays a pivotal role in reducing renal oxidative damage during haemolysis. PMID- 11865981 TI - Haptoglobin, an inflammation-inducible plasma protein. AB - Sterile tissue injury or infection initiates a local inflammatory response that mobilizes a systemic acute phase reaction resulting in, among other things, the induction of genes encoding the acute phase plasma proteins (APPs). In all vertebrates, a common set of APPs is increased and exerts essential protective functions. Haptoglobin (HP), one of the major APPs, acts as a high-affinity hemoglobin-binding protein and antioxidant. Liver is the major site of HP synthesis; however, regulated, low level expression is also detected in other organs. Induction of the Hp gene is mediated by interleukin-6-type cytokines and is synergistically enhanced by glucocorticoids. Growth stimulation of hepatic cells in vivo or in vitro suppresses the Hp gene-inducing effects of inflammatory cytokines. Receptors for IL-6 cytokines mediate induction of the Hp gene by the transcription factors signal transducer and activator of transcription-3 (STAT3) and CAAT/enhancer binding protein beta (C/EBPbeta), but attenuate the stimulation through co-activated STAT5 and mitogen-activated protein kinases, ERK-1 and ERK 2. The specificity by which the related cytokines, IL-6, oncostatin M, and leukemia inhibitory factor, regulate Hp gene transcription is determined by the profile of the cytokine receptor subunits expressed on the target cells and the relative extents by which these receptors activate the intracellular signaling pathways. The current hypothesis is that HP exerts an anti-inflammatory activity and that by the degree with which HP attenuates the inflammatory process, including the production of IL-6 cytokines, it determines the level and duration of acute phase expression of the Hp gene. PMID- 11865983 TI - Haptoglobin and malaria. AB - Haptoglobin gene knockout mice and wild-type controls were infected with Plasmodium berghei ANKA or Plasmodium chabaudi. The peak parasitaemia and parasite burden were higher in Hp-/- mice than in Hp+/+ mice. The increase in spleen weight following malaria infection was smaller in Hp-/- mice than in Hp+/+ animals. The occurrence of cerebral malaria in P. berghei ANKA infection was not different in Hp gene knockout mice and their controls. PMID- 11865982 TI - Haptoglobin and CD163: captor and receptor gating hemoglobin to macrophage lysosomes. AB - The plasma protein haptoglobin and the endocytic hemoglobin receptor HbSR/CD163 are key molecules in the process of removing hemoglobin released from ruptured erythrocytes. Hemoglobin in plasma is instantly bound with high affinity to haptoglobin--an interaction leading to the recognition of the complex by HbSR/CD163 and endocytosis in macrophages. The haptoglobin-dependent HbSR/CD163 scavenging system for hemoglobin clearance prevents toxic effects of hemoglobin in plasma and kidney and explains the decrease in the haptoglobin plasma concentration in patients with accelerated hemolysis. The HbSR/CD163 activity may be of quantitative importance for iron uptake in macrophages in general and for some iron-associated pathological processes, e.g. the atherogenesis-promoting oxidation of LDL leading to foam cell formation and apoptosis in the vessel wall. PMID- 11865984 TI - An indeterministic Monte Carlo technique for fast time of flight photon transport through optically thick turbid media. AB - A time-resolved indeterministic Monte Carlo (IMC) simulation technique is proposed for the efficient construction of the early part of the temporal point spread function (TPSF) of visible or near infrared photons transmitted through an optically thick scattering medium. By assuming a detected photon is a superposition of photon components, the photon is repropagated from a point in the original path where a significant delay in forward propagation occurred. A weight is then associated with each subsequently detected photon to compensate for shorter components. The technique is shown to reduce the computation time by a factor of at least 4 when simulating the sub-200 picosecond region of the TPSF and hence provides a useful tool for analysis of single photon detection in transillumination imaging. PMID- 11865985 TI - High frequency x-ray generator basics. AB - The purpose of this paper is to present basic functional principles of high frequency x-ray generators. The emphasis is put on physical concepts that determine the engineering solutions to the problem of efficient generation and control of high voltage power required to drive the x-ray tube. The physics of magnetically coupled circuits is discussed first, as a background for the discussion of engineering issues related to high-frequency power transformer design. Attention is paid to physical processes that influence such factors as size, efficiency, and reliability of a high voltage power transformer. The basic electrical circuit of a high frequency generator is analyzed next, with focus on functional principles. This section investigates the role and function of basic components, such as power supply, inverter, and voltage doubler. Essential electronic circuits of generator control are then examined, including regulation of voltage, current and timing of electrical power delivery to the x-ray tube. Finally, issues related to efficient feedback control, including basic design of the AEC circuitry are reviewed. PMID- 11865986 TI - Reference dosimetry calculations for neutron capture therapy with comparison of analytical and voxel models. AB - As clinical trials of Neutron Capture Therapy (NCT) are initiated in the U.S. and other countries, new treatment planning codes are being developed to calculate detailed dose distributions in patient-specific models. The thorough evaluation and comparison of treatment planning codes is a critical step toward the eventual standardization of dosimetry, which, in turn, is an essential element for the rational comparison of clinical results from different institutions. In this paper we report development of a reference suite of computational test problems for NCT dosimetry and discuss common issues encountered in these calculations to facilitate quantitative evaluations and comparisons of NCT treatment planning codes. Specifically, detailed depth-kerma rate curves were calculated using the Monte Carlo radiation transport code MCNP4B for four different representations of the modified Snyder head phantom, an analytic, multishell, ellipsoidal model, and voxel representations of this model with cubic voxel sizes of 16, 8, and 4 mm. Monoenergetic and monodirectional beams of 0.0253 eV, 1, 2, 10, 100, and 1000 keV neutrons, and 0.2, 0.5, 1, 2, 5, and 10 MeV photons were individually simulated to calculate kerma rates to a statistical uncertainty of <1% (1 std. dev.) in the center of the head model. In addition, a "generic" epithermal neutron beam with a broad neutron spectrum, similar to epithermal beams currently used or proposed for NCT clinical trials, was computed for all models. The thermal neutron, fast neutron, and photon kerma rates calculated with the 4 and 8 mm voxel models were within 2% and 4%, respectively, of those calculated for the analytical model. The 16 mm voxel model produced unacceptably large discrepancies for all dose components. The effects from different kerma data sets and tissue compositions were evaluated. Updating the kerma data from ICRU 46 to ICRU 63 data produced less than 2% difference in kerma rate profiles. The depth-dose profile data, Monte Carlo code input, kerma factors, and model construction files are available electronically to aid in verifying new and existing NCT treatment planning codes. PMID- 11865987 TI - Computerized diagnosis of breast lesions on ultrasound. AB - We present a computer-aided diagnosis (CAD) method for breast lesions on ultrasound that is based on the automatic segmentation of lesions and the automatic extraction of four features related to the lesion shape, margin, texture, and posterior acoustic behavior. Using a database of 400 cases (94 malignant lesions, 124 complex cysts, and 182 benign solid lesions), we investigate the marginal benefit of each feature in our CAD method and the performance of our CAD method in distinguishing malignant lesions from various classes of benign lesions. Finally, independent validation is performed on our CAD method. Eleven independent trials yielded an average Az value of 0.87 in the task of distinguishing malignant from benign lesions. PMID- 11865988 TI - Monte Carlo study of a highly efficient gas ionization detector for megavoltage imaging and image-guided radiotherapy. AB - The imaging characteristics of an arc-shaped xenon gas ionization chamber for the purpose of megavoltage CT imaging were investigated. The detector consists of several hundred 320 microm thick gas cavities separated by thin tungsten plates of the same thickness. Dose response, efficiency and resolution parameters were calculated using Monte Carlo simulations. The calculations were compared to measurements taken in a 4 MV photon beam, assuming that the measured signal in the chambers corresponds to the therein absorbed dose. The measured response profiles for narrow and broad incident photon beams could be well reproduced with the Monte Carlo calculations. They show, that the quantum efficiency is 29.2% and the detective quantum efficiency at zero frequency DQE(0) is 20.4% for the detector arc placed in focus with the photon source. For a detector placed out of focus, these numbers even increase. The efficiency of this kind of radiation detector for megavoltage radiation therefore surpasses the reported efficiency of existing detector technologies. The resolution of the detector is quantified with calculated and measured line spread functions. The corresponding modulation transfer functions were determined for different thicknesses of the tungsten plates. They show that the resolution is only slightly dependent on the plate thickness but is predominantly determined by the cell size of the detector. The optimal plate thickness is determined by a tradeoff between quantum efficiency, total signal generation and resolution. Thicker plates are more efficient but the total signal and the resolution decrease with plate thickness. In conclusion, a gas ionization chamber of the described type is a highly efficient megavoltage radiation detector, allowing to obtain CT images with very little dose for a sufficient image quality for anatomy verification. This kind of detector might serve as a model for a future generation of highly efficient radiation detectors. PMID- 11865989 TI - Intensity and energy modulated radiotherapy with proton beams: variables affecting optimal prostate plan. AB - Inverse planning for intensity- and energy-modulated radiotherapy (IEMRT) with proton beams involves the selection of (i) the relative importance factors to control the relative importance of the target and sensitive structures, (ii) an appropriate energy resolution to achieve an acceptable depth modulation, (iii) an appropriate beamlet width to modulate the beam laterally, and (iv) a sufficient number of beams and their orientations. In this article we investigate the influence of these variables on the optimized dose distribution of a simulated prostate cancer IEMRT treatment. Good dose conformation for this prostate case was achieved using a constellation of I factors for the target, rectum, bladder, and normal tissues of 500, 50, 15, and 1, respectively. It was found that for an active beam delivery system, the energy resolution should be selected on the basis of the incident beams' energy spread (sigmaE) and the appropriate energy resolution varied from 1 MeV at sigmaE = 0.0 to 5 MeV at sigmaE= 2.0 MeV. For a passive beam delivery system the value of the appropriate depth resolution for inverse planning may not be critical as long as the value chosen is at least equal to one-half the FWHM of the primary beam Bragg peak. Results indicate that the dose grid element dimension should be equal to or no less than 70% of the beamlet width. For this prostate case, we found that a maximum of three to four beam ports is required since there was no significant advantage to using a larger number of beams. However for a small number (< or = 4) of beams the selection of beam orientations, while having only a minor effect on target coverage, strongly influenced the sensitive structure sparing and normal tissue integral dose. PMID- 11865990 TI - Dosimetric properties of the new 125I BrachySeed model LS-1 source. AB - The BrachySeed model LS-1 is one of the latest in a series of new brachytherapy 125I seeds that have recently become available commercially for interstitial implants. The dosimetric properties of the seed were investigated analytically, experimentally, and by Monte Carlo simulation. Following the AAPM Task Group 43 formalism, the radial dose function, dose rate constant, and anisotropy parameters were determined. Experimental measurements were made in solid water equivalent phantoms using GafChromic MD-55-2 films, with correction for the low energy film response. Analyses were carried out from absolute measurements, as well as relative measurements against the Nycomed Amersham OncoSeed Model 6711, which also served to validate our experimental methodology. A small, but systematic difference in the absolute measurements was observed depending on the duration of the irradiation. Monte Carlo simulation was performed using the Integrated Tiger Series CYLTRAN code. We benchmarked the code by comparing the dose parameters of Model 6702 with published values. The radial dose function, g(r), of the Model LS-1 seed was computed at distances from 0.25 to 10 cm by analytical and Monte Carlo calculations with reasonably good agreement. The suggested dose rate constant, A, based on the Monte Carlo simulation is 0.90+/ 0.03 cGy h(-1) U(-1). This value is smaller than, but overlaps the experimental determination of 0.98+/-0.06 cGy h(-1) U(-1). The anisotropy function, F(r, theta), and anisotropy factor, phi(an)(r), compared favorably with those of the Model 6711. PMID- 11865991 TI - A comparative study of surface- and volume-based techniques for the automatic registration between CT and SPECT brain images. AB - Image registration of multimodality images is an essential task in numerous applications in three-dimensional medical image processing. Medical diagnosis can benefit from the complementary information in different modality images. Surface based registration techniques, while still widely used, were succeeded by volume based registration algorithms that appear to be theoretically advantageous in terms of reliability and accuracy. Several applications of such algorithms for the registration of CT-MRI, CT-PET, MRI-PET, and SPECT-MRI images have emerged in the literature, using local optimization techniques for the matching of images. Our purpose in this work is the development of automatic techniques for the registration of real CT and SPECT images, based on either surface- or volume based algorithms. Optimization is achieved using genetic algorithms that are known for their robustness. The two techniques are compared against a well established method, the Iterative Closest Point-ICP. The correlation coefficient was employed as an independent measure of spatial match, to produce unbiased results. The repeated measures ANOVA indicates the significant impact of the choice of registration method on the magnitude of the correlation (F = 4.968, p = 0.0396). The volume-based method achieves an average correlation coefficient value of 0.454 with a standard deviation of 0.0395, as opposed to an average of 0.380 with a standard deviation of 0.0603 achieved by the surface-based method and an average of 0.396 with a standard deviation equal to 0.0353 achieved by ICP. The volume-based technique performs significantly better compared to both ICP (p<0.05, Neuman Keuls test) and the surface-based technique (p<0.05, Neuman Keuls test). Surface-based registration and ICP do not differ significantly in performance. PMID- 11865992 TI - A slot-scanned photodiode-array/CCD hybrid detector for digital mammography. AB - We have developed a novel direct conversion detector for use in a slot-scanning digital mammography system. The slot-scan concept allows for dose efficient scatter rejection and the ability to use small detectors to produce a large-area image. The detector is a hybrid design with a 1.0 mm thick silicon PIN photodiode array (the x-ray absorber) indium-bump bonded to a CCD readout that is operated in time-delay integration (TDI) mode. Because the charge capacity requirement for good image quality exceeds the capabilities of standard CCDs, a novel CCD was developed. This CCD consists of 24 independent sections, each acting as a miniature CCD with eight rows for TDI. The signal from each section is combined off-chip to produce a full signal image. The MTF and DQE for the device was measured at several exposures and compared to a linear systems model of signal and noise propagation. Because of the scanning nature of TDI imaging, both the MTF(f) and DQE(f) are reduced along the direction of the scanning motion. For a 26 kVp spectrum, the DQE(0) was measured to be 0.75+/-0.02 for an exposure of 1.29 x 10(-5) C/kg (50 mR). PMID- 11865993 TI - Comparison of beam characteristics in intensity modulated radiation therapy (IMRT) and those under normal treatment condition. AB - In the step-and-shoot delivery of an IMRT plan with a Siemens Primus accelerator, radiation is turned off by desynchronizing the injector while the field parameters are being changed. When the machine is ready again a trigger pulse is sent to the injector to start the beam instantaneously. The objective of this study is to investigate the beam characteristics of the machine operating in the IMRT mode and to study the effect of the Initial Pulse Forming Network (IPEN) on the dark current. The central axis (CAX) output for a 10 x 10 cm2 field over the range 1-100 MU was measured with an ion chamber in a polystyrene phantom for both 6 and 15 MV x rays. Beam profiles were also measured over the range of 2-40 MU with the machine operating in the IMRT mode and compared with those in the normal mode. By adjusting the IPFN value, dark current radiation (DCR) was measured using ion chamber measurements. For both the normal and IMRT modes, dose versus MU is nonlinear in the range 1-5 MUs. Above 5 MU, dose varies linearly with MU for both 6 and 15 MV x rays. For stability of dose profiles, the 2 MU-IM group exhibit 20% variation from one subfield to another. The variation is about 5% for the 8 MU-IM group and <5% for 10 MU and higher. The results are similar in the normal treatment mode. With the IPFN at >80% of the PFN value, a spurious radiation associated with dark current at approximately 0.7% of the dose at isocenter for a 10 x 10 cm2 field is detected during the "PAUSE" state of the accelerator for 15 MV x rays. When the IPFN is lowered to <80% of the PFN value, no DCR is detected. For 6 MV x rays, no measurable DCR was detected regardless of the IPFN setting. PMID- 11865994 TI - Reducing dose calculation time for accurate iterative IMRT planning. AB - A time-consuming component of IMRT optimization is the dose computation required in each iteration for the evaluation of the objective function. Accurate superposition/convolution (SC) and Monte Carlo (MC) dose calculations are currently considered too time-consuming for iterative IMRT dose calculation. Thus, fast, but less accurate algorithms such as pencil beam (PB) algorithms are typically used in most current IMRT systems. This paper describes two hybrid methods that utilize the speed of fast PB algorithms yet achieve the accuracy of optimizing based upon SC algorithms via the application of dose correction matrices. In one method, the ratio method, an infrequently computed voxel-by voxel dose ratio matrix (R = D(SC)/D(PB)) is applied for each beam to the dose distributions calculated with the PB method during the optimization. That is, D(PB) x R is used for the dose calculation during the optimization. The optimization proceeds until both the IMRT beam intensities and the dose correction ratio matrix converge. In the second method, the correction method, a periodically computed voxel-by-voxel correction matrix for each beam, defined to be the difference between the SC and PB dose computations, is used to correct PB dose distributions. To validate the methods, IMRT treatment plans developed with the hybrid methods are compared with those obtained when the SC algorithm is used for all optimization iterations and with those obtained when PB-based optimization is followed by SC-based optimization. In the 12 patient cases studied, no clinically significant differences exist in the final treatment plans developed with each of the dose computation methodologies. However, the number of time-consuming SC iterations is reduced from 6-32 for pure SC optimization to four or less for the ratio matrix method and five or less for the correction method. Because the PB algorithm is faster at computing dose, this reduces the inverse planning optimization time for our implementation by a factor of 2 to 8 compared with pure SC optimization, without compromising the quality or accuracy of the final treatment plan. PMID- 11865995 TI - Improvement of computerized mass detection on mammograms: fusion of two-view information. AB - Recent clinical studies have proved that computer-aided diagnosis (CAD) systems are helpful for improving lesion detection by radiologists in mammography. However, these systems would be more useful if the false-positive rate is reduced. Current CAD systems generally detect and characterize suspicious abnormal structures in individual mammographic images. Clinical experiences by radiologists indicate that screening with two mammographic views improves the detection accuracy of abnormalities in the breast. It is expected that the fusion of information from different mammographic views will improve the performance of CAD systems. We are developing a two-view matching method that utilizes the geometric locations, and morphological and textural features to correlate objects detected in two different views using a prescreening program. First, a geometrical model is used to predict the search region for an object in a second view from its location in the first view. The distance between the object and the nipple is used to define the search area. After pairing the objects in two views, textural and morphological characteristics of the paired objects are merged and similarity measures are defined. Linear discriminant analysis is then employed to classify each object pair as a true or false mass pair. The resulting object correspondence score is combined with its one-view detection score using a fusion scheme. The fusion information was found to improve the lesion detectability and reduce the number of FPs. In a preliminary study, we used a data set of 169 pairs of cranio-caudal (CC) and mediolateral oblique (MLO) view mammograms. For the detection of malignant masses on current mammograms, the film-based detection sensitivity was found to improve from 62% with a one-view detection scheme to 73% with the new two-view scheme, at a false-positive rate of 1 FP/image. The corresponding cased-based detection sensitivity improved from 77% to 91%. PMID- 11865996 TI - Physics concepts that cannot be explained from a clinical context should be omitted in physics courses for radiologists. For the proposition. PMID- 11865998 TI - Comment on "Self-absorption correction for 32P, 198Au, and 188Re stents: dose point kernel calculations versus Monte Carlo". PMID- 11865999 TI - Tiotropium in the management of chronic obstructive pulmonary disease. PMID- 11866000 TI - A breakthrough in the functional evaluation of the inspiratory muscle pump. PMID- 11866001 TI - A long-term evaluation of once-daily inhaled tiotropium in chronic obstructive pulmonary disease. AB - Currently available inhaled bronchodilators used as therapy for chronic obstructive pulmonary disease (COPD) necessitate multiple daily dosing. The present study evaluates the long-term safety and efficacy of tiotropium, a new once-daily anticholinergic in COPD. Patients with stable COPD (age 65.2+/-8.7 yrs (mean+/-SD), n=921) were enrolled in two identical randomized double-blind placebo-controlled 1-yr studies. Patients inhaled tiotropium 18 microg or placebo (mean screening forced expiratory volume in one second (FEV1) 1.01 versus 0.99 L, 39.1 and 38.1% of the predicted value) once daily as a dry powder. The primary spirometric outcome was trough FEV1 (i.e. FEV1 prior to dosing). Changes in dyspnoea were measured using the Transition Dyspnea Index, and health status with the disease-specific St. George's Respiratory Questionnaire and the generic Short Form 36. Medication use and adverse events were recorded. Tiotropium provided significantly superior bronchodilation relative to placebo for trough FEV1 response (approximately 12% over baseline) (p<0.01) and mean response during the 3 h following dosing (approximately 22% over baseline) (p<0.001) over the 12 month period. Tiotropium recipients showed less dyspnoea (p<0.001), superior health status scores, and fewer COPD exacerbations and hospitalizations (p<0.05). Adverse events were comparable with placebo, except for dry mouth incidence (tiotropium 16.0% versus placebo 2.7%, p<0.05). Tiotropium is an effective, once daily bronchodilator that reduces dyspnoea and chronic obstructive pulmonary disease exacerbation frequency and improves health status. This suggests that tiotropium will make an important contribution to chronic obstructive pulmonary disease therapy. PMID- 11866002 TI - Magnetic resonance analysis of abnormal diaphragmatic motion in patients with emphysema. AB - The purpose of this study was to quantitatively evaluate paradoxical diaphragmatic motion using magnetic resonance (MR) imaging. A total of 27 subjects were examined, including 12 normal young adults, six control individuals, and nine patients with emphysema. With subjects in the supine position, 30 sequential sagittal MR images of the entire right lung were obtained during tidal and deep slow breathing. Diaphragmatic movement between sequential images was estimated as the displacement area and the total diaphragmatic movement in a respiratory cycle was calculated. The paradoxical motion of the diaphragm, representing the inverted movement to increase or decrease lung area, since paradoxical movement ratio (Mpr=(total paradoxical diaphragmatic movement/total diaphragmatic movement)x100), was evaluated. In patients with emphysema, paradoxical diaphragmatic motion was observed on MR images during deep breathing. The mean Mpr in emphysematous patients during deep breathing was 10+/ 4%, which was significantly higher than 0.5+/-0.2% in young adults (p<0.05), and 1.2+/-0.6% in aged-matched controls (p<0.05). The present results indicate that magnetic resonance images could be used to detect paradoxical diaphragmatic motion in patients with emphysema. PMID- 11866003 TI - A novel clinical test of respiratory muscle endurance. AB - Impaired respiratory muscle endurance (RME) could reduce exercise tolerance and contribute to ventilatory failure. The aim of the present study was to develop a clinically-feasible method to measure RME using negative-pressure inspiratory threshold loading. It was hypothesized that endurance time (tlim) could be predicted by normalizing oesophageal pressure-time product (PTP) per total breath cycle (PTPoes) for maximum oesophageal pressure (Poes,max); the load/capacity ratio. The corresponding mouth pressures, PTPmouth and Pmouth,max were also measured. The RME test was performed on 30 healthy subjects exposed to the same target pressure (70% of Poes,max). Eight patients with systemic sclerosis/interstitial lung disease were studied to assess the validity and acceptability of the technique. Normal subjects showed a wide intersubject variation in tlim (coefficient of variation, 69%), with a linear relationship demonstrated between log tlim and PTPoes/Poes,max (r=0.88). All patients with systemic sclerosis/interstitial lung disease had normal respiratory muscle strength, but six out of eight had a reduction in RME. In conclusion, endurance time can be predicted from the load/capacity ratio, over a range of breathing strategies; this relationship allows abnormal respiratory muscle endurance to be detected in patients. Oesophageal and mouth pressure showed a close correlation, thus suggesting that the test could be applied noninvasively. PMID- 11866004 TI - Misuse of corticosteroid metered-dose inhaler is associated with decreased asthma stability. AB - This study assessed whether the improper use of pressurized metered-dose inhalers (pMDIs) is associated with decreased asthma control in asthmatics treated by inhaled corticosteroids (ICS). General practitioners (GPs) included consecutive asthmatic outpatients treated by pMDI-administered ICS and on-demand, short acting beta2-agonists. They measured an asthma instability score (AIS) based on daytime and nocturnal symptoms, exercise-induced dyspnoea, beta2-agonist usage, emergency-care visits and global perception of asthma control within the preceding month; the inhalation technique of the patient also was assessed. GPs (n=915) included 4,078 adult asthmatics; 3,955 questionnaires were evaluable. pMDI was misused by 71% of patients, of which 47% was due to poor coordination. Asthma was less stable in pMDI misusers than in good users (AIS: 3.93 versus 2.86, p<0.001). Among misusers, asthma was less stable in poor coordinators (AIS: 4.38 versus 3.56 in good coordinators, p<0.001). To conclude, misuse of pressurized metered-dose inhalers, which is mainly due to poor coordination, is frequent and associated with poorer asthma control in inhaled corticosteroid treated asthmatics. This study highlights the importance of evaluating inhalation technique and providing appropriate education in all patients, especially before increasing inhaled corticosteroid dosage or adding other agents. The use of devices which alleviate coordination problems should be reinforced in pressurized metered-dose inhaler misusers. PMID- 11866005 TI - European Community Respiratory Health Survey calibration project of dosimeter driving pressures. AB - Two potential sources of systematic variation in output from Mefar dosimeters, the system used in the European Community Respiratory Health Survey (ECRHS) study have been evaluated: individual nebulizer characteristics and dosimeter driving pressure. Output variation from 366 new nebulizers produced in two batches for the second ECRHS were evaluated, using a solute tracer method, at a fixed driving pressure. The relationship between dosimeter driving pressure was then characterized and between-centre variation in dosimeter driving pressure was evaluated in an Internet-based survey. A systematic difference between nebulizers manufactured in the two batches was identified. Batch one had a mean+/-SD output of 7.0+/-0.8 mg x s(-1) and batch two, 6.3+/-0.7 mg x s(-1) (p<0.005). There was a wide range of driving pressures generated by Mefar dosimeters as set, ranging between 70-245 kPa, with most outside the quoted manufacturer's specification of 180+/-5%. Nebulizer output was confirmed as linearly related to dosimeter driving pressure (coefficient of determination (R2)=0.99, output=0.0377 x driving pressure-0.4151). The range in driving pressures observed was estimated as consistent with a variation of about one doubling in the provocative dose causing a 20% fall in forced expiratory volume in one second. Systematic variation has been identified that constitutes potentially significant confounders for between centre comparisons of airway responsiveness in the European Community Respiratory Health Survey, with the dosimeter driving pressure representing the most serious issue. This work confirms the need for appropriate quality control of both nebulizer output and dosimeter driving pressure, in laboratories undertaking field measurements of airway responsiveness. In particular, appropriate data on driving pressures need to be collected and factored into between-centre comparisons. Comprehensive collection of such data to optimize quality control is practicable and has been instigated by the organizing committee for the European Community Respiratory Health Survey II. PMID- 11866006 TI - Fluticasone induces apoptosis in peripheral T-lymphocytes: a comparison between asthmatic and normal subjects. AB - Apoptosis is an important mechanism allowing inflammation to be limited. Glucocorticoids are the most effective anti-inflammatory agents in asthma therapy and induce cell apoptosis. Since T-lymphocytes are critically involved in airway inflammation in asthma, the effects of fluticasone propionate (FP) on apoptosis in unstimulated and in interleukin (IL)-2 stimulated peripheral blood T lymphocytes (PBTs) isolated from 14 normal and 19 mild-to-moderate asthmatic subjects were evaluated. Apoptosis was evaluated by: deoxyribonucleic acid (DNA) fragmentation electrophoresis, DNA content, annexin V binding, apoptosis related markers (Fas, B-cell lymphona leukaemia-2 (Bcl-2), Bax, and CD25), and by electron microscopy. FP induced apoptosis in unstimulated PBTs of normal and asthmatic subjects in a time-dependent fashion. In asthma, this effect was associated with a significant decrease of Bcl-2 expression, and with an increase of Bax/Bcl-2 ratio. In PBTs of asthmatics, FP also reduced Fas and CD25 expression. Moreover, in IL-2-stimulated PBTs from both asthmatics and normal subjects, FP was able to induce apoptosis and to reduce Bcl-2, Fas and CD25 expression, whereas negligible effects were detected on Bax expression. This study shows that the glucocorticosteroid, fluticasone, increases apoptosis and modulates expression of apoptosis-related markers in unstimulated and in interleukin-2 stimulated T-lymphocytes. This points towards a potential mechanism by which fluticasone exerts its anti-inflammatory effects. PMID- 11866007 TI - Anti-Chlamydophila immunoglobulin prevalence in sarcoidosis and usual interstitial pneumoniae. AB - Sarcoidosis and usual interstitial pneumoniae (UIP) are diseases of unknown aetiology affecting the lower respiratory tract. Although there are a number of studies investigating the causal role of these disorders, no micro-organism could be identified as the causal agent. The high incidence of Chlamydophila pneumoniae infections associated with lung injury encouraged the present investigations to screen patients with sarcoidosis and with UIP for their Chlamydophila-specific immune response. Thirty-nine patients with sarcoidosis, 26 patients with UIP and 34 controls were tested for the prevalence of Chlamydophila-specific antibodies in bronchoalveolar lavage fluids (BALF) and sera. Samples were tested for the presence of antibodies in a genus-specific test for Chlamydophila lipopolysaccharide (LPS) and in a species-specific test for C. pneumoniae. This study revealed a significantly higher prevalence of Chlamydophila LPS-specific immunoglobulin (Ig)-G in the BALF of sarcoidosis patients (36.8%) compared to controls (8.8%) and patients with UIP (12.0%). Similar findings were observed in sera. The prevalence of C. pneumoniae-specific antibodies in BALF was significantly higher in sarcoidosis patients for IgG and IgA (IgG: 74.4%; IgA: 46.2%) and in UIP for IgG (IgG: 50.0%; IgA: 11.5%) compared to controls (IgG: 14.7%; IgA: 14.7%). The elevated prevalence of Chlamydophila-specific antibodies in sarcoidosis patients might implicate Chlamydophila as a causal agent. However, considering the high prevalence of Chlamydophila antibodies in the healthy population, the data presented might reflect Chlamydophila co-infections in pre injured lungs seen in these patients. PMID- 11866009 TI - Dose-finding and 24-h monitoring for efficacy and safety of aerosolized Nacystelyn in cystic fibrosis. AB - The aim of the present studies was to investigate the tolerability and activity of a novel mucolytic drug, Nacystelyn (NAL), for the treatment of cystic fibrosis (CF) lung disease. In study 1, involving 10 CF patients, the main objective was to determine the tolerability and potential efficacy of a range of single doses of NAL in comparison to a placebo, in order to establish an optimal dose for further testing. On five consecutive scheduled treatment days, patients inhaled either from two (4 mg) to eight puffs (16 mg) of a single dose of NAL from the range, administered in an open-label fashion, or 12 puffs of active NAL (24 mg) versus 12 puffs of placebo, administered in a randomized double-blind fashion. Pulmonary function data were unaffected and clinically-adverse effects were limited to wheezing in some patients that inhaled 12 puffs of either placebo or active drug. Subsequent rheological analysis of their sputum showed a dose dependent decrease in sputum viscoelasticity, accompanied by a decrease in sputum solids content and an increase in chloride and sodium concentrations. In study 2, involving 12 CF patients, the clinical safety and mucolytic activity of a single dose of NAL was monitored over 24 h. On different scheduled treatment days, 7 days apart, patients inhaled a single dose of 12 puffs of active NAL (24 mg) or 12 puffs of placebo drug in a randomized, double-blind sequence, with sputum samples taken at intervals before and after inhalation. Mucus rigidity decreased following NAL inhalation, with the maximum effect observed at 4 h; the 1-, 2- and 4-h NAL rheology results were significantly different from placebo. No adverse effects were observed. The drug was well tolerated in both studies. Sputum results were predictive of improved clearability by ciliary and cough transport mechanisms. PMID- 11866008 TI - Clinical significance of histological classification of idiopathic interstitial pneumonia. AB - Patients with idiopathic interstitial pneumonias (IIPs) can be subdivided into groups based on the histological appearance of lung tissue obtained by surgical biopsy. The quantitative impact of histological diagnosis, baseline factors and response to therapy on survival has not been evaluated. Surgical lung biopsy specimens from 168 patients with suspected IIP were reviewed according to the latest diagnostic criteria. The impact of baseline clinical, physiological, radiographic and histological features on survival was evaluated using Cox regression analysis. The predictive value of honeycombing on high-resolution computed tomography (HRCT) as a surrogate marker for usual interstitial pneumonia (UIP) was examined. The response to therapy and survival of 39 patients treated prospectively with high-dose prednisone was evaluated. The presence of UIP was the most important factor influencing mortality. The risk ratio of mortality when UIP was present was 28.46 (95% confidence interval (CI) 5.5-148.0; p=0.0001) after controlling for patient age, duration of symptoms, radiographic appearance, pulmonary physiology, smoking history and sex. Honeycombing on HRCT indicated the presence of UIP with a sensitivity of 90% and specificity of 86%. Patients with nonspecific interstitial pneumonia were more likely to respond or remain stable (9 of 10) compared to patients with UIP (14 of 29) after treatment with prednisone. Patients remaining stable had the best prognosis. The risk ratio of mortality for stable patients compared to nonresponders was 0.32 (95% CI 0.11 0.93; p=0.04) in all patients and 0.33 (95% CI 0.12-0.96; p=0.04) in patients with UIP. The histological diagnosis of usual interstitial pneumonia is the most important factor determining survival in patients with suspected idiopathic interstitial pneumonia. The presence of honeycombing on high-resolution computed tomography is a good surrogate for usual interstitial pneumonia and could be utilized in patients unable to undergo surgical lung biopsy. Patients with nonspecific interstitial pneumonia are more likely to respond or remain stable following a course of prednisone. Patients remaining stable following prednisone therapy have the best prognosis. PMID- 11866010 TI - Once-daily tobramycin in the treatment of adult patients with cystic fibrosis. AB - The aim of this study was to test the equivalence of once- and thrice-daily dosing with tobramycin by comparing efficacy and safety in adult patients with cystic fibrosis. Sixty adult patients with an acute respiratory exacerbation were randomized to receive either 10 mg x kg(-1) tobramycin once-daily or 3.3 mg x kg( 1) tobramycin thrice-daily. Primary efficacy and safety endpoints were defined as changes in respiratory function and changes in renal function and hearing. Both groups showed a significant increase in respiratory function without a clinically significant change in renal function. For changes in forced vital capacity % predicted and serum potassium and magnesium levels, equivalence was demonstrated. For the variables forced expiratory volume in one second and forced mid expiratory flow % pred and serum creatinine levels, there was insufficient power to demonstrate equivalence. One patient in each group showed bilateral impairment in pure tone audiogram after treatment. This study demonstrated significant clinical improvement with both once- and thrice-daily tobramycin dosing. Equivalence between the two regimens was shown for some, but not all primary endpoints. Once-daily dosing should be used with careful monitoring of safety and efficacy until large multicentre studies confirm these encouraging results. PMID- 11866011 TI - Noninvasive ventilation in cystic fibrosis patients with acute or chronic respiratory failure. AB - The experience of using noninvasive ventilation (NIV) in 113 adult cystic fibrosis (CF) patients with chronic respiratory failure, during episodes of acute deterioration in respiratory function is reported. The patients aged 15-44 yrs were divided into three groups. Group A consisted of 65 patients (median forced expiratory volume in one second (FEV1)/forced vital capacity (FVC) 0.7/1.4 L) who were on a lung transplant waiting list. Group B consisted of 25 patients (median FEV1/FVC 0.7/1.4 L) who were being evaluated for lung transplantation. Group C consisted of 23 patients (median FEV1/FVC 0.6/1.2 L) who were not being considered for lung transplantation. The mean duration of NIV support for groups A, B and C was 61 (range: 1-600) days, 53 (1-279) days and 45 (0.5-379) days respectively. Twenty-three patients in group A subsequently received lung transplantation and 12 of these patients had a median survival of 39 months postsurgery. Thirty-nine patients died and three awaited transplantation. Five patients in group B received a transplant four of whom survived; thirteen patients died and seven awaited transplantation. Twenty patients in group C died. Noninvasive ventilation improved hypoxia but failed to correct hypercapnia in these cystic fibrosis patients. Noninvasive ventilation is useful in the treatment of acute episodes of respiratory failure in cystic fibrosis patients with end-stage lung disease who have been accepted, or are being evaluated, for lung transplantation. For these patients, there is a possibility of prolonging life if they are successfully treated for their acute episode of respiratory failure until transplantation. In this group, treatment is not merely prolonging the process of dying. PMID- 11866012 TI - Management of lower respiratory tract infections by French general practitioners: the AIR II study. Analyse Infections Respiratoires. AB - The Analyse Infections Respiratoires (AIR) II study is a prospective, multicentre survey of the management of lower respiratory tract infections in patients aged 15-65 yrs by general practitioners (GPs) in France. To obtain real-time data recording, practitioners were required to submit an anonymous copy of their drug prescriptions. They were then interviewed over the telephone about the patients' sociodemographic data, signs and symptoms, as well as their presumptive diagnosis and the investigations they had decided upon. GPs (n=3,144) reported 5,469 evaluable cases. Pneumonia accounted for 9.6% of diagnoses, acute exacerbations of chronic bronchitis 14.9% and acute bronchitis 72.5%. The symptomatology covered an extremely wide range of clinical features, which, although statistically different in terms of incidence, overlapped to a large extent across diagnoses. By contrast, hospitalization, investigations or referral to a specialist were much more prevalent in pneumonia, although still very infrequent in general terms (0.5, 1.2 and 10.8%, respectively). Antibiotics were prescribed in 96.5% of patients, with minor differences between diagnoses. However, other medications such as nonsteroid, anti-inflammatory drugs, steroids, nonspecific antitussives and bronchial liquefiers accounted for two-thirds of the prescriptions. This study demonstrates the lower respiratory tract infections encountered by general practitioners are usually mild. However, antibiotic prescription was more systematic than in previous studies and the prescription of nonspecific symptomatic treatments was twice as frequent. General practitioners did not perform additional examinations or refer on a regular basis. There was a high prescription rate for symptomatic treatment. PMID- 11866013 TI - Pleural-fluid myeloperoxidase in complicated and noncomplicated parapneumonic pleural effusions. AB - The diagnostic accuracy of myeloperoxidase (MPO) in pleural fluid, for differentiating between complicated and noncomplicated parapneumonic pleural effusions (PPE) evaluated prospectively. Seventy patients aged >18 yrs with PPE (36 complicated and 34 noncomplicated) were studied after admission to a tertiary referral teaching hospital. MPO concentration was measured in plasma and pleural fluid using a double-antibody competitive radioimmunoassay. The concentrations of MPO in complicated and noncomplicated PPE were compared using a Mann-Whitney U test and multiple logistic regression models were used to predict the odds that an effusion was complicated. MPO pleural-fluid concentrations were significantly higher in complicated than in noncomplicated PPE. After excluding purulent effusions, pleural-fluid MPO was the marker that best differentiated between the two types of PPE: the area under the receiver operating characteristic curve was 0.912, the sensitivity was 87.5% and the specificity was 85.1% at a cut-point limit of 3.000 microg x L(-1). The authors concluded that the concentration of pleural-fluid myeloperoxidase helps to differentiate between nonpurulent complicated and noncomplicated parapneumonic pleural effusions. PMID- 11866014 TI - Bronchial bacterial colonization in patients with resectable lung carcinoma. AB - The pattern and clinical implications of bronchial bacterial colonization have been widely investigated in patients with chronic lung disease, particularly chronic obstructive pulmonary disease. The main aim of this study was to determine the frequency and risk factors for bronchial colonization in lung cancer patients who have undergone surgical resection. Forty-one patients with resectable lung cancer (22 (54%) active smokers, 52+/-23 pack-yrs) with a mean forced expiratory volume in one second of 80+/-16% predicted, were studied with bilateral protected specimen brush and lung tissue biopsy during the surgical procedure. Quantitative bacterial culture, susceptibility tests and histological examination of samples were performed. Bronchial colonization with > or = 1 potential pathogenic micro-organism was found in 17 of 41 (41%) patients. The most frequent strains isolated were: Haemophilus influenzae (35%), Streptococcus pneumoniae (13%) and Pseudomonas spp. (9%). The risk factors for bronchial colonization were central location of the tumour (odds ratio (OR)=9.2, confidence interval (CI) 95%=2.1-39.6, p=1.003) and increased body mass index (OR=1.6, CI 95%=1.2-2.2, p=0.005). The frequency of postoperative infectious pulmonary complications was low (five cases (12%)) and no relationship was observed with bronchial colonization. Patients with resectable lung carcinoma had a high rate of bronchial colonization (41%), mainly with potential pathogenic microorganisms. The independent risk factors for colonization in these patients were central location of the tumour and a high body mass index. PMID- 11866015 TI - Mechanisms and markers of airway inflammation in cystic fibrosis. AB - Airway inflammation is now recognized as a major factor in the pathogenesis of lung disease in cystic fibrosis (CF). Its most characteristic feature is a marked and persistent influx into the airways of neutrophils, which damage the lung by releasing noxious mediators, such as reactive oxygen species and proteolytic enzymes. Recent studies suggest that inflammation occurs very early and may even happen in the absence of infection. Furthermore, links between CF transmembrane conductance regulator dysfunction and both infection and inflammation are postulated; dysregulation of cytokine production and abnormal epithelial host defences have been regarded as causes of sustained inflammation. Bronchoalveolar lavage and the evaluation of neutrophils and inflammatory mediators provide the most accurate picture of airway inflammation. Routine bronchoscopy with bronchoalveolar lavage, however, is unpleasant for the patient and usually is of no immediate benefit to the management of individual cases. Therefore, surrogate markers collected by noninvasive procedures would be of great assistance in the follow-up of cystic fibrosis patients. Several markers have been evaluated in the sputum, serum and urine of cystic fibrosis patients and related to the degree of airway inflammation. Long-term studies are needed to confirm their potential clinical utility and specificity, and to determine which can be used clinically to monitor disease outcome and efficacy of treatment. PMID- 11866017 TI - ERS/ATS statement on interventional pulmonology. European Respiratory Society/American Thoracic Society. PMID- 11866016 TI - Respiratory viral infections as promoters of allergic sensitization and asthma in animal models. AB - Respiratory virus infections can trigger exacerbations of asthma and may also contribute to allergic sensitization to aeroallergens and the development of asthma. Conversely, atopy may predispose to more severe virus-induced airway disease. The animal models reviewed in this article support the hypothesis that respiratory virus infections can promote allergic sensitization and the development of asthma. Respiratory viruses can prevent induction of tolerance and enhance sensitization to inhaled allergens resulting in increased airway inflammation and airway hyperresponsiveness. Probable mechanisms involved in this enhanced sensitization are increased permeability of the airway mucosa to allergens and recruitment of dendritic cells to the respiratory epithelium during acute infection. Factors involved in augmenting the consequences of allergic airway sensitization appear to be T-cells, especially CD8+ T-cells as regulators of this process, interleukin-5 as a pivotal cytokine for eosinophilic airway inflammation and eosinophils themselves as effector cells triggering airway hyperresponsiveness. Depending on the timing of allergen exposure, respiratory virus infections which elicit a significant type 1 T-helper cell cytokine response may also downregulate allergic sensitization. Respiratory virus infections in animals previously sensitized to aeroallergens result in prolonged increases in inflammation and airway responsiveness, indicating that critical interactions between immune responses to allergen sensitization and the responses to infection can lead to more severe disease. Taken together, animal models have proved valuable in generating a number of plausible pathogenetic concepts, and can be used to address a host of unresolved questions regarding the immunology of respiratory virus infections, allergic sensitization and asthma. PMID- 11866018 TI - Identification of a new cystic fibrosis transmembrane regulator mutation in a severely affected patient. AB - By using a combination of multiplex polymerase chain reaction and allele-specific labelled probes, the oligo-ligation assay is designed to detect known cystic fibrosis transmembrane regulator mutations. This study shows that this assay may also be useful to detect new mutations. The second child of a family of Bosnic origin showed all the symptoms of intestinal and pulmonary manifestations of cystic fibrosis. No signal could be obtained for the allele-specific probe 1898+1G>A. This could be explained by a nearby localized sequence change that prevented polymerase chain reaction primers or oligonucleotide probes from binding to the target sequence. Indeed, sequence analysis revealed a new 1894G>T exchange (Glu587Stop). Both parents and the healthy brother carried this mutation. Thus, the index patient was homozygous for 1894G>T, which was inherited from both parents. PMID- 11866019 TI - Haemorrhagic hypersensitivity pneumonitis due to naphthylene-1,5-diisocyanate. AB - Symptoms of hypersensitivity pneumonitis and massive pulmonary haemorrhage occurred in a 24-yr-old male shortly after occupational exposure to naphthylene 1,5-diisocyanate (NDI). The present examination was performed approximately 1-yr after the initial life-threatening haemoptysis and following an uneventful recovery after resection of the middle lobe, which had been identified bronchoscopically as the bleeding source. Histological re-examination of the lung was compatible with hypersensitivity pneumonitis. After a chamber challenge with NDI (5 parts per billion (ppb) for 10 min, 10 ppb for 110 min), rales were heard in both lungs, and a fall in vital capacity and partial pressure of arterial oxygen as well as a rise in body temperature were documented. Isocyanate-specific immunoglobulin-G antibodies could not be detected in the patient's serum, possibly due to the long period without exposure to isocyanates. The authors conclude that naphthylene-1,5-diisocyanate may cause immunological pulmonary haemorrhage. The underlying disease is consistent with hypersensitivity pneumonitis and may be triggered by low concentrations of the diisocyanate. PMID- 11866020 TI - Chlorine exposure and the upper respiratory tract. PMID- 11866021 TI - Usefulness of transcutaneous Doppler jugular venous echo to predict pulmonary hypertension in COPD patients. PMID- 11866022 TI - Contemporary methods in hypnotic preparation for childbirth. AB - Hypnosis is a viable adjunct to any medical procedure and is not intended to replace conventional medical techniques. In recent years, many of us who practice hypnosis have seen a re-emergence of interest in mind-body approaches to health care. Hypnotic methods for preparation for childbirth are a logical contribution to that mind-body perspective. PMID- 11866023 TI - The use of pre-, intra-, and posthypnotic suggestion in anesthesia and surgery. AB - While under hypnosis, patients can be taught to alter their psychophysiological functions. With this ability to alter these functions, patients can overcome the anxiety associated with surgery. Patients with high anxiety often experience more depression, can have increased complications, need more anesthesia and medication, have suppressed immune function, and often take longer to heal. The purpose of this article is to review the research literature related to the use of hypnosis in preparing the patient for surgery and to present 2 approaches used by the authors to prepare patients for surgery. The first approach is used when there is enough time to condition the patient, and the second approach is used when the anesthetist meets the patient shortly before the surgery is to begin and there is no time to induce formal trance. PMID- 11866024 TI - Clinical applications of acupuncture in anesthesia practice. AB - The most widely successful use of acupuncture in Western medicine has been in the treatment of chronic and intractable pain syndromes. Thus, it is especially important for the nurse anesthetist who is the practice of chronic pain management to be familiar with this treatment choice. This article provides the nurse anesthetist with basic information about the practice of acupuncture, the patient who may ask for acupuncture, application of segmental acupuncture techniques, and legislative and licensure issues. PMID- 11866025 TI - Healing pathways through energy work in the perianesthesia care setting. AB - Energy-medicine therapy such as healing touch is a powerful way to promote relaxation and enhance the healing process. Healing touch is a sacred healing art and a way of caring in which practitioners use their hands as channels to assess and balance the energy field that encircles the body in order to promote the innate ability to heal. A collection of energy-based treatment modalities are used to assess and treat the human energy system. The energy system that is life is influenced by healing touch, which is used extensively in the nursing profession. This energy-medicine therapy is used in all areas of nursing. This article discusses the concepts of healing touch, the human energy field, and applications of healing touch in professional practice in the perianesthesia setting. PMID- 11866026 TI - The use of hypnosis in a primary care setting. AB - An aware clinician using the modality of hypnosis either themselves or induced by a trained practitioner will find many indications on a daily basis. The application can be used for control of symptoms such as nausea or pain or, in the case of some clinical conditions, as a primary or adjunctive therapy. An overview of the commonly occurring opportunities for the use of hypnosis is presented in this article. PMID- 11866027 TI - Hypnosis principles and applications: an adjunct to health care. AB - Hypnosis has existed since the beginning of humankind, and is a part of everyday life. It is a valuable addition to the methods and techniques available to all health care-providers, as well as a safe and uncomplicated method used to enhance patient health care. It is simply a state of complete physical and mental relaxation which produces an altered state of consciousness acceptable to suggestions. It is characterized by an increased ability to produce desirable changes in habit patterns, motivation, self-image, lifestyle, and personal health. PMID- 11866028 TI - Death of a journal: lost opportunities, new challenges, or both? AB - The announced cessation of publication of CRNA: The Clinical Forum for Nurse Anesthetists brings into focus the problems that confront smaller subscription based professional journals in a print or paper mode. We are in an age in which there is a high level of competition for peoples' time and attention, in an environment where there is a danger of information overload. Electronic media are competing with print media, and there are advantages and disadvantages to both. This is not only within the realm of news, sports, and entertainment, but professional information as well. Debates will abound as to how best, and in what medium, research should be made available to professions and the public. Unfortunately, the revelations in the past few years of the poor state of published research in our print journals, despite peer review, makes it more difficult to advocate them in a climate where the shift may well be toward electronic journals. Although many print journals have also gone online, they usually limit the viewership to abstracts or a few articles. Some of thejournals that have been created as online journals and have their articles peer reviewed have not been in existence long enough to examine the extent to which the peer review system is any better than for print journals. It is exceedingly important that research and its conclusions that make its way into either print or online journals are reliable and valid before we apply them to our practice. Relying on individual readers to make that determination is problematic, because most readers are not that astute in sophisticated research methods and statistics. With the loss of this journal, CRNA opportunities for research and commentary publication are lost. It will produce new challenges to interested CRNAs who choose to balance this loss with new opportunities. PMID- 11866029 TI - Measuring communication and social skills in a high security forensic setting using the behavioural status index. AB - Assessing patient functioning in the areas of communication and social skills is a core area of practice within psychiatric nursing. Difficulties within these areas can often represent the root of a number of presenting problems. Objective assessment can be difficult without a validated assessment schema. The Behavioural Status Index (BSI) offers such a system of baseline assessment and longitudinal monitoring as a basis for treatment, further specialised assessment, or measurement of outcome during and after interventions. This paper introduces some basic theory and describes the function and purpose of the BSI. This is followed by data analysis for the BSI communications and social skills sub-scale. Data were collected, using a repeated measures method by primary nurses, from a sample of 503 individual patients in two high security mental health hospitals. Results are reported for descriptive statistics, and factor analysis; and differences between the independent groups of Mental Health Act 1983 classification, ward dependency and gender. Data trends are indicative of clinically interesting relationships. A distinct factorial structure emerged suggesting groupings of assertive-interpersonal, non-verbal, social-perceptive and para-linguistic behaviours. Results are reported to be underpinning current European studies. PMID- 11866030 TI - Depression among immigrant Mexican women and Southeast Asian refugee women in the U. S. AB - Although immigrants and refugees share the experience of adapting to a new country, life experiences and circumstances surrounding leaving their homelands are vastly different. The most salient difference is their motivation for leaving. Immigrants typically leave their homeland to seek improved economic opportunities and/or to join other family members. Refugees leave their homeland under the threat of injury or loss of life due to political or religious persecution and severe deprivation of basic life necessities. Since the decision to migrate is often viewed as a positive change for immigrant women in comparison to refugee women, mental health problems may be under -detected. The researchers will describe the prevalence of depression in two of the largest groups of migrant women in the U.S., immigrant Mexican women (N=220) and refugee Southeast Asian women (N=163). The purpose of this paper is to compare and contrast life circumstances that may impact on the prevalence of depression in both groups of women. The issues presented are important for nurses internationally who assess and design interventions for immigrant and refugee populations of women. PMID- 11866031 TI - Measuring risk in a high security forensic setting through the behavioural status index. AB - This paper reports data analysis for the Behavioural Status Index risk sub-scale. Data were collected, using a repeated measures method by primary nurses, from a sample of 503 individual patients in two high security mental health hospitals. Results are reported for descriptive statistics, and factor analysis; and differences between the independent groups of Mental Health Act 1983 classification, ward dependency and gender. Data trends are indicative of clinically interesting relationships. A distinct factorial structure emerged suggesting groupings of attentional, violent, destructive, iconic-compensatory and escapist behaviours. Results are reported to be underpinning current European studies. PMID- 11866032 TI - Heme oxygenase as an intrinsic defense system in vascular wall: implication against atherogenesis. AB - Recent developments in our understanding of the atherosclerotic process and factors that trigger ischemic cardiovascular disease have led to the consideration of antioxidative responses or exogenous antioxidants, which are proposed to inhibit multiple proatherogenic and prothrombotic events in arterial wall. Heme oxygenases (HO), an enzyme essential for heme degradation, have been shown to have such antioxidative properties via the production of bile pigments, carbon monoxide and ferritin induction. We have demonstrated that mildly oxidized LDL markedly induces HO-1, an inducible form of HO, in human aortic endothelial and smooth muscle cell cocultures and that its induction results in the attenuation of monocyte chemotaxis induced by mildly oxidized LDL. We also confirmed abundant expression of HO-1 in human, murine and rabbit atherosclerotic lesions. By modulating HO activities in LDL-receptor knockout mice and Watanabe heritable hyperlipidemic rabbits during their atherosclerotic lesion developments, anti-atherogenic properties of HO have demonstrated as judged by the quantitative analyses of atherosclerotic lesion formation. HO expression was inversely correlated with the levels of plasma and tissue lipid peroxides. HO also influenced on nitric oxide pathway. These observations may suggest that HO, induced during atherosclerotic process, functions as an intrinsic protective pathway in vascular wall. PMID- 11866033 TI - Four strains of spontaneously hyperlipidemic (SHL) mice: phenotypic distinctions determined by genetic backgrounds. AB - Spontaneously hyperlipidemic (SHL) mice are Japanese wild mice (KOR) with disruption of the apolipoprotein E (Apo E) gene. These mice (KOR-Apoe(shl)) are superhypercholesterolemic and develop severe xanthoma, but their atherosclerosis is relatively mild compared with Apo E knockout mice. First, we tested whether this distinction is due to additional mutation of the Apoc1 and/or Apoc2 genes in KOR-Apoe(shl). Southern blot analysis, but found no gross disruption of these genes. Next, we tested whether the phenotypic distinction is due to differences in the genetic background. To this end, we established three lines of congenic SHL mice with a genetic background of C57BL/6, BALB/c or C3H/He, and named them, respectively, C57BL/6.KOR-Apoe(shl) (B6.KOR-Apoe(shl)), BALB/c.KOR-Apoe(shl) (C.KOR-Apoe(shl)) and C3H/He.KOR-Apoe(shl) (C3.KOR-Apoe(shl)). Hypercholesterolemia was most severe in KOR-Apoe(shl) followed the by others as follows; KOR-Apoe(shl)>>C3.KOR-Apoe(shl)>C.KOR-Apoe(shl)>B6.KOR-Apoe(shl). In contrast, atherosclerosis was most severe in B6.KOR Apoe(shl) followed by the others: B6.KOR-Apoe(shl)>C.KOR-Apoe(shl)>>C3.KOR-Apoe(shl)> or =KOR-Apoe(shl). This order, however, did not match that in xanthoma, which was highly prominent in KOR-Apoe(shl) but mild in B6.KOR-Apoe(shl), C.KOR-Apoe(shl) and C3.KORApoe(shl). This order, however, did not match that in xanthoma, which was highly prominant in KOR-Apoe(shl) but mild in B6.KOR-Apoe(shl), C.KOR-Apoe(shl) and C3.KOR-Apoe(shl). These distinctions suggest that the severity of each of the phenotypes is determined by distinct genetic backgrounds which probably are composed of polymorphism of lipid metabolism-related proteins. We found that apolipoprotein A-I is decreased in each SHL strain and polymorphic between B6.KOR Apoe(shl) and the other strains examined. This polymorphism may be related to the most severe atherosclerosis observed in B6.KOR-Apoe(shl). It is most likely that combination of such polymorphisms is due to the genetic background accountable for phenotype distinctions. PMID- 11866034 TI - No association found between the Ala54Thr polymorphism of FABP2 gene and obesity and obesity with dyslipidemia in Japanese schoolchildren. AB - To investigate whether the Ala54Thr polymorphism of the fatty acid binding protein 2 gene is associated with obesity and obesity with dyslipidemia in Japanese schoolchildren, we analyzed 370 children with morbid obesity and 463 control children of normal weight. The allele frequencies did not differ significantly between the control group and the morbidly obese group. The odds ratio (95% confidence interval CI) in obesity of the The54 allele was 1.0 (0.9 1.3). There were no significant differences in obesity index and metabolic characteristics between the two groups. The odds ratio (95% CI) in dyslipidemia of the Thr54 allele was 1.1 (0.8-1.4) in the morbidly obese group. Our data suggested that Ala54Thr polymorphism of the FABP2 gene is not a major contributing factor for obesity and obesity with dyslipidemia in Japanese children. PMID- 11866035 TI - Comparison of direct methods and HPLC for the measurement of HDL- and LDL cholesterol with ultracentrifugation. AB - Although ultracentrifugation is the gold standard for lipoprotein analysis, inexpensive and easy direct methods for HDL- and LDL-cholesterol (C) have recently been developed. In this study, we compared representative methods of lipoprotein analysis, namely, ultracentrifugation, direct assay methods, and HPLC, to measure LDL- and HDL-C. A good correlation was observed between HDL-C by ultracentrifugation and HDL-C by direct methods or HPLC. A good correlation was also observed between LDL-C (d1.006-1.063) by ultracentrifugation and LDL-C by direct methods or HPLC. Although the correlation between LDL-C (d1.019-1.063) by ultracentrifugation and LDL-C by direct methods was also good, the correlation coefficient was significantly decreased, suggesting that 'LDL-C' by direct methods correlates better with LDL-C (d1.006-1.063) than LDL-C (d1.019-1.063) by ultracentrifugation. Although the correlation between IDL-C (d1.006-1.019) by ultracentrifugation and the difference in LDL-C by direct methods and LDL-C (d1.019-1.063) by ultracentrifugation was investigated, no significant correlation was observed. The IDL-C contained in LDL-C (d1.006-1.063) varied from 2-28%. In homozygous CETP-deficient and LCAT-deficient subjects, the dissociation was marked. It is crucial to understand that 'LDL-C' in the Guidelines for the Diagnosis and Treatment of Hyperlipidemias in Adults by the Japanese Atherosclerosis Society should be considered to be LDL-C (d1.006-1.063) and that 'LDL-C' by direct assay methods means LDL-C (d1.006-1.063) by ultracentrifugation. PMID- 11866036 TI - Association of mild hyperhomocysteinemia with aortic calcification in hypercholesterolemic patients. AB - Homocysteine is considered to be an independent risk factor for atherosclerosis. Experimental animal models of hyperhomocysteinemia show aortic calcification, suggesting that this disorder is associated with aortic calcification in humans. A total of 28 patients with hyperlipidemia were enrolled into this study. The degree of aortic calcification at the level of the bifurcation and 1 cm proximal to the bifurcation was assessed by computed tomography of the aorta and the association between calcification of the aorta and the plasma level of homocysteine was then analyzed. The mean plasma homocysteine level in 28 patients was 8.7 microM. They were divided into 2 groups, high homocysteine level group (HHL; homocysteine level >8.7 microM) and low homocysteine level group (LHL; homocysteine level < = 8.7 microM). The degree of aortic calcification at the level of the bifurcation differed significantly between the two groups (19.1% vs. 10.5%; p < 0.01). We found that mild hyperhomocysteinemia was associated with aortic calcification, which suggests that interventions to reduce the plasma level of homocysteine may also reduce the severity of aortic calcification. PMID- 11866037 TI - Significance of a polymorphism (G-->A transition) in the -75 position of the apolipoprotein A-I gene promoter on serum high density lipoprotein-cholesterol levels in Japanese hyperlipidemic subjects. AB - High density lipoprotein-cholesterol (HDL-C) levels are inversely related to the incidence of coronary artery disease. We studied the influence of a G(-75)-->A transition in the promoter of the apolipoprotein (apo) A-I gene, a major protein component of HDL, on serum HDL-C levels in hyperlipidemic subjects. Seventy three hyperlipidemic subjects with serum levels of high HDL-C (HDL-C > or = 70 mg/dl, Group H) were compared with hyperlipidemic subjects with levels of HDL-C between 40 and 70 mg/dl (Group N) and those with HDL-C < 40 mg/dl (Group L). Group H showed a higher incidence (45.2%) of low plasma cholesteryl ester transfer protein (CETP) activity than Groups N (9.1%) and L (5.3%) (p < 0.001). Group H had a higher incidence of the G(-75)-->A transition (0.275) than Groups N (0.117, p < 0.05) and L (0.056, p < 0.01), among subjects with normal CETP activities. The HDL-C levels in subjects with the transition (84 +/- 16 mg/dl) were higher than those in subjects without the transition (56 +/- 12 mg/dl) (p < 0.05). These data suggest that a G(-75)-->A transition of the apo A-I gene promoter, in addition to the common mutation of CETP gene, contributes to high HDL-C levels among hyperlipidemic patients in Japan. PMID- 11866038 TI - Meningeal enhancement on magnetic resonance imaging in 15 dogs and 3 cats. AB - A retrospective study of 15 dogs and three cats was done to characterize the appearance of meningeal enhancement on magnetic resonance (MR) images of the brain, and to correlate this appearance with its underlying cause. Two patterns of meningeal enhancement (pial and dural) were identified. Enhancement of the pia mater was evident in four dogs and one cat, while enhancement of the dura mater was seen in 11 dogs and 2 cats. A variety of causes of meningeal enhancement were identified, including bacterial and cryptococcal meningitis, plasmacytic meningitis with associated subdural fluid accumulation, granulomatous meningoencephalomyelitis, inflammation secondary to otitis interna, feline infectious peritonitis, and neoplasia. The present study confirms that pial or dural meningeal enhancement may be present on MR images of the brain of dogs or cats in association with a variety of central nervous system diseases. A larger prospective study is required to further establish the incidence of specific patterns of meningeal enhancement seen in association with specific diseases. PMID- 11866039 TI - Computed tomographic anatomy of the canine inner and middle ear. AB - A series of high-resolution computed x-ray tomography (CT) images of the normal canine middle and inner ear are presented to serve as a reference for optimal interpretation of clinical CT images of animals with diseases affecting this region. PMID- 11866040 TI - Osteoid osteoma in a dog. PMID- 11866041 TI - Cretinism in a North American black bear (Ursus americanus). AB - Congenital hyperplastic goiter and cretinism were documented in a 16 month-old male North American black bear (Ursus americanus). The cub was captured at approximately 8 months of age and maintained for an additional 8 months in captivity. Clinical signs included growth retardation, clumsiness, and facial dysmorphism. Hypothyroidism was documented by determining serum triiodothyronine (T3) and thyroxine (T4) levels. Lysosomal storage disease was ruled out by measuring various lysosomal enzyme activities. Serologic, radiographic, computed tomographic, necropsy, and histopathologic findings were consistent with congenital hypothyroidism and cretinism. PMID- 11866043 TI - An unusual case of upper respiratory obstruction in a horse. PMID- 11866042 TI - Superparamagnetic iron oxide-enhanced magnetic resonance imaging of the liver in beagle dogs. AB - The role of superparamagnetic iron oxide as a tissue-specific contrast medium has been established in humans, especially for hepatic imaging. Superparamagnetic iron oxide particles exhibit a tissue-specific biodistribution to the reticuloendothelial system, where they predominantly shorten transverse T2 relaxation time. Most hepatic tumors lack Kupffer cells; therefore, the T2 of tumors remains virtually unchanged after administration of superparamagnetic iron oxide. The resulting loss of signal intensity from the liver, with unchanged tumor signal intensity, increases lesion-to-liver contrast. In this study, MR images were acquired with fast gradient echo recalled at steady state (FGRE) in five Beagle dogs before and after injection of superparamagnetic iron oxide. The effect of superparamagnetic iron oxide on signal intensity of the liver with time was assessed. A signal intensity decrease of 65.7+/-10.0% was detected at 20 minutes, and it continued to decrease until the last time point of MR scanning (200 minutes). The liver intensity of all dogs dropped to half its value after 20 minutes. The effect of motion was minimized by breath holding. Superparamagnetic iron oxide did not have any adverse effects on the dogs. PMID- 11866044 TI - Congenital facial infiltrative lipoma in a calf. AB - The clinical and pathologic findings related to an infiltrative facial lipoma in a 5-month-old female Holstein Friesian calf are reported. The tumor was congenital, increased in size with time, and deformed the left side of the face and cranial bones. The clinical condition of the calf was good; the only detectable abnormalities were poor weight gain and decreased motility of the left upper lip. Clinical and radiologic data indicated the mass was not removable. Muscle and lymph node invasion by tumor tissue were detected histologically. Infiltrative lipoma is a rare variety of lipoma that has been reported in dogs, cats, horses, and humans. Although its cytologic characteristics are those of a benign tumor, in this patient the invasiveness of the neoplasm was associated with poor prognosis. PMID- 11866045 TI - Computed tomographic findings in 35 dogs with nasal aspergillosis. AB - The purpose of this study was to describe the computed tomographic (CT) features of nasal aspergillosis in dogs. Initial (n = 35) and follow-up (n = 12) CT images were available from 35 dogs. The most commonly encountered CT findings were (1) moderate to severe cavitary destruction of the turbinates with presence of a variable amount of abnormal soft tissue in the nasal passages, (2) non-specific thickening of the mucosa adjacent to the inner surface of bones of the frontal sinus, maxillary recess and nasal cavity and, (3) thickened reactive bone. The findings were consistent with a disease initially affecting one nasal cavity then progressing into the ipsilateral frontal sinus, the contralateral nasal cavity and the contralateral frontal sinus. Two dogs with associated nasal foreign body had a more localized invasion of the nasal cavity. Attenuation values and contrast enhancement were not specific. With follow-up examinations, a reduction in the amount of abnormal soft tissue was observed in all dogs except one, but this reduction could not be quantified. PMID- 11866046 TI - Needle-tract implantation following us-guided fine-needle aspiration biopsy of transitional cell carcinoma of the bladder, urethra, and prostate. AB - Localized tumor implantation of the ventral abdominal wall was found at 2, 5, and 8 months following percutaneous ultrasound-guided fine-needle aspiration biopsy (FNAB) of transitional carcinoma of the bladder, urethra, or prostate in 3 dogs. To our knowledge this complication has not been reported in dogs following FNAB. Despite the rarity of needle-tract implantation, the potential for this complication with transitional cell carcinomas is apparently not negligible and warrants consideration. We recommend traumatic urethral catheterization to obtain a cytologic diagnosis of potential transitional cell carcinomas of the lower urinary tract or prostate whenever possible until more information becomes available. However, needle-track implantation is so rare that it should not influence the decision to perform a percutaneous FNAB if the urethra cannot be catheterized. PMID- 11866047 TI - Transorbital echoencephalography in cattle. AB - The purpose of this study was establishment of the criteria of transorbital echoencephalography in cattle and experimental applications to bovine practice. Quantitative investigations using magnetic resonance (MR) imaging revealed that this examination could be applied to cattle under 3 months of age. The method of transducer positioning was established in Japanese Black (J.B.) and filial (F1) cattle (turning caudally at an angle of about 16 degrees and dorsally at an angle of about 23 degrees) or in Holstein cattle (turning caudally at an angle of about 20 degrees and dorsally at an angle of about 21 degrees). Examinations in clinical calves revealed that the cerebral parenchyma and the lateral ventricle could be detected antemortem or postmortem. In this study, the diagnoses of hydrocephalus or hydranencephaly was possible using antemortem transorbital echoencephalography. Transorbital echoencephalography was especially useful as the imaging method for bovine hydranencephaly. PMID- 11866048 TI - A lipoma of the extensor tendon sheaths in a horse. AB - This report describes the identification and surgical removal of a lipoma from the extensor tendon sheaths of a horse. To the authors' knowledge, this is the first report of a lipoma of the extensor tendon sheaths of a horse. Ultrasonographic evaluation was crucial to patient management, providing information regarding the soft tissue mass characteristics, tumor margins and synovial involvement prior to surgical exploration. Surgical removal was performed and was curative. PMID- 11866049 TI - Images from the 2001 ACVR certifying examination: CT/MR elective section. PMID- 11866050 TI - Image from the 2001 radiation oncology certifying examination: clinical aspects of radiation oncology, including image interpretation. PMID- 11866051 TI - Integrated microfluidic electrophoresis system for analysis of genetic materials using signal amplification methods. AB - An isothermal signal amplification technique for specific DNA sequences, known as cycling probe technology (CPT), was performed within a microfluidic chip. The presence of DNA from methicillin-resistant Staphylococcus aureus was determined by signal amplification of a specific DNA sequence. The microfluidic device consisted of four channels intersecting to mix the sample and reagents within 55 s, as they were directed toward the reactor coil by electrokinetic pumping. The 160-nL CPT reactor occupied approximately 220 mm2. Gel-free capillary electrophoresis separation of the biotin- and fluorescein-labeled probe from the probe fragments was performed on-chip following the on-chip reaction. An off-chip CPT reaction, with on-chip separation gave a detection limit of 2 fM (0.03 amol) target DNA and an amplification factor of 85,000. Calibration curves, linear at <5% probe fragmentation, obeyed a power law relationship with an argument of 0.5 [target] at higher target DNA concentrations for both on-chip and off-chip CPT reaction and analysis. An amplification factor of 42,000 at 250 fM target (25,000 target molecules) was observed on-chip, but the reaction was approximately 4 times less sensitive than off-chip under the conditions used. Relative SD values for on-chip CPT were 0.8% for the peak migration times, 9% for the area of intact probe peak, and 8% for the fragment/probe peak area ratio. PMID- 11866052 TI - "Internal residue loss": rearrangements occurring during the fragmentation of carbohydrates derivatized at the reducing terminus. AB - Rearrangement reactions involving migration of fucose and, occasionally, other residues have been found in the CID spectra of [M + H]+ and [M + 2H]2+ ions, but not [M + Na]+ ions, generated from several O-linked carbohydrates and milk sugars derivatized at their reducing termini with aromatic amines such as 2 aminobenzamide. Such rearrangements, which are similar to those reported by other investigators from several underivatized carbohydrates and glycosides, cause an apparent loss of sugar residues from within a carbohydrate chain and can produce ambiguous results during spectral interpretation. A mechanism, involving initial protonation of the amine nitrogen atom of the derivative, is proposed to account for the formation of the observed ions. PMID- 11866053 TI - Characterization of transthyretin variants in familial transthyretin amyloidosis by mass spectrometric peptide mapping and DNA sequence analysis. AB - Transthyretin (TTR) is a 127-amino acid residue transport protein. In plasma, TTR exists as a tetramer and binds the hormone thyroxine and the retinol-binding protein-vitamin A complex. Amino acid substitutions in TTR are hypothesized to destabilize the tetramer and cause the protein to form intermediates that self associate into amyloid fibrils. Familial transthyretin amyloidosis (ATTR) is associated with extracellular deposition of wild-type TTR, its variants or fragments as amyloid fibrils in various tissues and organs. A definitive diagnosis of ATTR depends on the detection and identification of TTR variants. Electrospray ionization (ESI) and matrix-assisted laser desorption/ionization (MALDI) time-of-flight (TOF) mass spectrometry (MS), in combination with trypsin digestion, have been shown to be powerful tools in characterizing TTR variants. Typically, TTR or its tryptic digest is analyzed by MALDI-TOF MS, liquid chromatography ESI MS, or both. Analysis of tryptic digests by MALDI-TOF MS does not provide enough sequence coverage in TTR to identify all possible modifications. To improve sequence coverage, aliquots of immunoprecipitated TTR samples were digested with trypsin, lysyl endopeptidase Lys-C, or endoproteinase Asp-N. Identification of the peptides from each digest by MALDI-TOF MS provided preliminary information about the sites and mass shifts due to amino acid substitutions from genetic mutations and to posttranslational modifications. The location and identity of the modifications in the variant proteins were then confirmed by tandem mass spectrometry, accurate mass measurements, and direct DNA sequence analysis. Using these methodologies, we achieved 100% sequence coverage. The detection of two nonpathologic variants (Thr119Met and Gly6Ser) and four pathologic variants (Phe64Leu, Asp38Ala, Phe44Ser, and previously unreported Trp41Leu) are described as illustrations of this approach. PMID- 11866054 TI - Sol-gel capillary microextraction. AB - Sol-gel capillary microextraction (sol-gel CME) is introduced as a viable solventless extraction technique for the preconcentration of trace analytes. To our knowledge, this is the first report on the use of sol-gel-coated capillaries in analytical microextraction. Sol-gel-coated capillaries were employed for the extraction and preconcentration of a wide variety of polar and nonpolar analytes. Two different types of sol-gel coatings were used for extraction: sol-gel poly(dimethylsiloxane) (PDMS) and sol-gel poly(ethylene glycol) (PEG). An in house-assembled gravity-fed sample dispensing unit was used to perform the extraction. The analysis of the extracted analytes was performed by gas chromatography (GC). The extracted analytes were transferred to the GC column via thermal desorption. For this, the capillary with the extracted analytes was connected to the inlet end of the GC column using a two-way press-fit fused silica connector housed inside the GC injection port. Desorption of the analytes from the extraction capillary was performed by rapid temperature programming (at 100 degrees C/min) of the GC injection port. The desorbed analytes were transported down the system by the helium flow and further focused at the inlet end of the GC column maintained at 30 degrees C. Sol-gel PDMS capillaries were used for the extraction of nonpolar and moderately polar compounds (polycyclic aromatic hydrocarbons, aldehydes, ketones), while sol-gel PEG capillaries were used for the extraction of polar compounds (alcohols, phenols, amines). The technique is characterized by excellent reproducibility. For both polar and nonpolar analytes, the run-to-run and capillary-to-capillary RSD values for GC peak areas remained under 6% and 4%, respectively. The technique also demonstrated excellent extraction sensitivity. Parts per quadrillion level detection limits were achieved by coupling sol-gel CME with GC-FID. The use of thicker sol-gel coatings and longer capillary segments of larger diameter (or capillaries with sol-gel monolithic beds) should lead to further enhancement of the extraction sensitivity. PMID- 11866055 TI - Fundamentals of capillary electrochromatography: migration behavior of ionized sample components. AB - The mechanism of separating charged species by capillary electrochromatography (CEC) was modeled with the conditions of ideal/linear chromatography by using a simple random walk. The most novel aspect of the work rests with the assumption that in sufficiently high electric field ionized sample components can also migrate in the adsorbed state on the ionized surface of the stationary phase. This feature of CEC leads to the introduction of three dimensionless parameters: alpha, reduced mobility of a sample component with the electrosmotic mobility as the reference; beta, the CEC retention factor; and gamma, the ratio of the electrophoretic migration velocity and the velocity of surface electrodiffusion. Since the interplay of retentive and electrophoretic forces determines the overall migration velocity, the separation mechanism in CEC is governed by the relative importance of the above parameters. The model predicts conditions under which the features of the CEC system engender migration behavior that manifests itself in a relatively narrow elution window and in a gradient like elution pattern in the separation of peptides and proteins by using pro forma isocratic CEC. It is believed that such elution patterns, which resemble those obtained by the use of external gradient of the eluent, are brought about by the formation of an internal gradient in the CEC system that gave rise to concomitant peak compression. The peculiarities of CEC are discussed in the three operational modalities of the technique: co-current, countercurrent, and co-counter CEC. The results suggest that CEC, which is often called "liquid chromatography on electrophoretic platform" is an analytical tool with great potential in the separation of peptides and proteins. PMID- 11866056 TI - Effect of analyte adsorption on the electroosmotic flow in microfluidic channels. AB - The predictability and constancy over time of the electroosmotic flow in microchannels is an important consideration in microfluidic devices. A common cause for alteration of the flow is the adsorption of analytes to channel walls, for example, during capillary electrophoresis of proteins. It is shown that certain experimental data, published by Towns and Regnier (Towns, J. K; Regnier, F. E. Anal. Chem. 1992, 64, 2473-2478.), on the anomalous elution times for proteins in capillary electrophoresis can be explained using a simple model for analyte adsorption that uses a result first reported by Anderson and Idol (Anderson, J. L.; Idol, W. K Chem. Eng. Commun. 1985, 38, 93-106.) on the electroosmotic flux in capillaries with axial variations in zeta-potential. It is suggested that it might be possible to use such a model to dynamically correct for altered elution times in capillary electrophoretic devices. PMID- 11866057 TI - Phospholipid bilayer coatings for the separation of proteins in capillary electrophoresis. AB - The double-chained, zwitterionic phospholipid 1,2-dilauroyl-sn phosphatidylcholine (DLPC, C12) was investigated for its use as a wall coating for the prevention of protein adsorption in capillary electrophoresis. DLPC forms a semipermanent coating at the capillary wall, which allows excess phospholipid to be removed from the capillary prior to electrophoretic separation. A DLPC coated capillary allowed for the separation of both cationic and anionic proteins with efficiencies as high as 1.4 million plates/m. Migration time reproducibility was less than 1.3% RSD from run to run and less than 4.0% RSD from day to day. Protein recovery was as high as 93%. Cationic and anionic proteins could be separated over a pH range of 3-10, all yielding good efficiencies (N up to 1 million plates/m). The chain length of the phospholipid affected the performance of the wall coating. The C10 analogue of DLPC (DDPC) did not form a coating on the capillary wall while the C14 analogue of DLPC (DMPC) formed a stable coating that prevented protein adsorption to the same extent as its C12 counterpart. PMID- 11866059 TI - Determination of polybrominated diphenyl ethers and polychlorinated biphenyls in human adipose tissue by large-volume injection-narrow-bore capillary gas chromatography/electron impact low-resolution mass spectrometry. AB - A new analytical method has been developed for the quantification of polybrominated diphenyl ethers (PBDEs) in human adipose tissue samples. After Soxhlet extraction and a cleanup procedure using two successive solid-phase extraction cartridges containing acid silica and acid silica: neutral silica:deactivated basic alumina (from top to bottom), detection can been achieved by narrow-bore (0.10-mm i.d.) capillary gas chromatography/electron impact low-resolution mass spectrometry using a large-volume injection technique. Using narrow-bore capillaries, it is possible to analyze complex mixtures in a short time (up to 10 min), saving 50% or more of the analysis time of conventional columns while maintaining a similar resolution power. The method allows the determination of five major PBDE congeners (BDE 28, 47, 99, 100, and 153) at concentrations below 1 ng/g lipid weight. Detection limits in the selected ion mode varied between 0.05 and 0.30 ng/g lipid weight, depending on the degree of bromination. The sensitivity of this method can compete with low resolution mass spectrometry with electron capture ionization, while a much better selectivity is obtained. Levels of PBDEs in 20 Belgian human adipose tissue samples ranged between 2.18 and 11.70 ng/g lipid weight and were similar to previously reported values from Europe. PMID- 11866058 TI - Electrochromatography in microchips: reversed-phase separation of peptides and amino acids using photopatterned rigid polymer monoliths. AB - A microfabricated glass chip containing fluidic channels filled with polymer monolith has been developed for reversed-phase electrochromatography. Acrylate based porous polymer monoliths were cast in the channels by photopolymerization to serve as a robust and uniform stationary phase. UV light-initiated polymerization allows for patterning of polymer stationary phase in the microchip, analogous to photolithography, using a mask and a UV lamp for optimal design of injection, separation, and detection manifolds. The monoliths are cast in situ in less than 10 min, are very reproducible with respect to separation characteristics, and allow easy manipulation of separation parameters such as charge, hydrophobicity, and pore size. Moreover, the solvent used to cast the polymer enables electroosmotic flow, allowing the separation channel to be conditioned without need for high-pressure pumps. The microchip was used for separation of bioactive peptides and amino acids labeled with a fluorogenic dye (naphthalene-2,3-dicarboxaldehyde) followed by laser-induced fluorescence detection using a Kr+ ion laser. The microchip-based separations were fast (six peptides in 45 s), efficient (up to 600,000 plates/m), and outperformed the capillary-based separations in both speed and efficiency. We have also developed a method for complete removal of polymer from the channels by thermal incineration to regenerate the glass chips. PMID- 11866060 TI - Fundamental studies of liquid chromatography at the critical condition using enhanced-fluidity liquids. AB - The improvement in the analysis of telechelic polymer matrixes continues to be a pursuit for many scientists of varying disciplines. This quest for a new technique has led to the continued development of liquid chromatography at the critical condition (LCCC) or liquid chromatography at the critical adsorption point (LC-CAP). LCCC allows for the isolation of one area of the polymer matrix so that other areas of the polymer can be probed with size-exclusion or adsorptive chromatographic modes. Although this technique has been successfully applied to the analysis of telechelic polymers, the practice of LCCC can be difficult. These difficulties include finding and maintaining a solvent system appropriate for the practice of LCCC as well as deterioration of peak shape once the system is operating at the LCCC mode. Because of the specificity of the mobile phase required for the practice of LCCC, the work is routinely practiced by premixing solvents. Previous work with enhanced-fluidity liquid mobile phases demonstrated that these mobile phases removed many of the aforementioned challenges associated with working at the LCCC mode. These mobile phases utilize both pressure and temperature variation in order to maintain the specific solvent strength necessary for the LCCC work. This work studies the coupling and optimization of enhanced-fluidity, EF, liquid mobile phases for LCCC. Several EF LCCC systems, differing in mobile phase composition, temperature, and pressure, were routinely established, resulting in the effective practice of critical chromatography. The practice of LCCC with on-line mobile phase preparation is demonstrated using commercially available instrumentation. Finally, EF-LCCC is used to analyze triblock and diblock copolymers. PMID- 11866061 TI - An automated on-line multidimensional HPLC system for protein and peptide mapping with integrated sample preparation. AB - A comprehensive on-line two-dimensional 2D-HPLC system with integrated sample preparation was developed for the analysis of proteins and peptides with a molecular weight below 20 kDa. The system setup provided fast separations and high resolving power and is considered to be a complementary technique to 2D gel electrophoresis in proteomics. The on-line system reproducibly resolved approximately 1000 peaks within the total analysis time of 96 min and avoided sample losses by off-line sample handling. The low-molecular-weight target analytes were separated from the matrix using novel silica-based restricted access materials (RAM) with ion exchange functionalities. The size-selective sample fractionation step was followed by anion or cation exchange chromatography as the first dimension. The separation mechanism in the subsequent second dimension employed hydrophobic interactions using short reversed-phase (RP) columns. A new column-switching technique, including four parallel reversed-phase columns, was employed in the second dimension for on-line fractionation and separation. Gradient elution and UV detection of two columns were performed simultaneously while loading the third and regenerating the fourth column. The total integrated workstation was operated in an unattended mode. Selected peaks were collected and analyzed off-line by MALDI-TOF mass spectrometry. The system was applied to protein mapping of biological samples of human hemofiltrate as well as of cell lysates originating from a human fetal fibroblast cell line, demonstrating it to be a viable alternative to 2D gel electrophoresis for mapping peptides and small proteins. PMID- 11866062 TI - An optical fiber chemical sensor for mercury ions based on a porphyrin dimer. AB - The synthesis of a novel fluoroionophore, 5-p-[[4-(10',15',20'-triphenyl-5' porphinato) phenyloxyl]-1-butyloxyl]phenyl-10,15,20-triphenylporphine (DTPP), and its application for preparation of a Hg(II)-sensitive optical fiber chemical sensor are described. The response of the sensor is based on the fluorescence quenching of DTPP by coordination with Hg(II). The porphyrin dimer-based sensor shows a linear response toward Hg(II) in the concentration range 5.2 x 10(-7)-3.1 x 10(-4) mol x L(-1), with a working pH range from 2.4 to 8.0. The sensor shows excellent selectivity for Hg(II) over transition metal cations including Cd(II), Co(II), Cu(II), Ni(II), Pb(II), Zn(II), and Fe(III). As a sensing agent, the porphyrin dimer shows obviously better fluorescence response characteristics toward Hg(II) compared to porphyrin monomer or metalloporphyrin. The effect of the composition of the sensor membrane was studied, and the experimental conditions were optimized. The sensor has been used for determination of Hg(II) in water samples. PMID- 11866063 TI - Sequential injection 90Sr determination in environmental samples using a wetting film extraction method. AB - A sequential injection procedure involving a flow-reversal wetting-film extraction method for the determination of the radionuclide 90Sr has been developed. The methodology is based on the coating of the inner walls of an open tubular reactor with a film prepared from a 0.14 M 4,4'(5')-bis(tert butylcyclohexano)-18-crown-6 (BCHC) solution in 1-octanol, which allows the selective isolation of strontium from the sample matrix. Selection of the optimum extractant diluent attending its physical properties, investigation of the extraction kinetics features, and choice of the proper elution procedure are discussed in detail in this paper. The noteworthy aspects of using a wetting-film phase instead of a solid-phase material described to date in the literature are the reduction of crown ether consumption and the simplification of both the operational sequence and the automation of the extractant-phase renewal between consecutive samples, which is of interest to avoid analyte carryover and reduction of the resin capacity factor caused by irreversible interferences. The proposed method has been successfully applied to different spiked environmental samples (water, milk, and soil), with 90Sr total activities ranging between 0.07 and 0.30 Bq, measured using a low-background proportional counter. The standard deviation of the automated analytical separation procedure is lower than 3% (n = 10), and the 90Sr isolation process under the studied conditions may be carried out with a yield up to 80%. PMID- 11866064 TI - Label-free parallel screening of combinatorial triazine libraries using reflectometric interference spectroscopy. AB - The parallel reflectometric interference spectroscopy is presented as a label free optical detection method. A new setup was adapted to accommodate sample carriers in a 96-well microplate. It allows for the first time simultaneous plate imaging by a CCD camera for the parallel detection of specific biomolecular interaction in the microplate wells at heterogeneous phase using direct optical monitoring. The detection of binding events with time resolution enables a highly parallel functional biomolecular interaction analysis (BIA). The combination of this new screening setup with combinatorial solid-phase synthesis is performed in the wells of glass-bottom microplates to accomplish the synthesis and the screening platform within one device. As a model system for a solid-phase substance library, synthesis of a triazine library and the subsequent BIA with four different antibodies were carried out. The presented setup enables a time resolution of 18 s with a total screening time of less than 35 min including baseline adjustment, BIA, and regeneration of the screening device for 96 samples in parallel. The binding studies reveal a fast classification of the different monoclonal and polyclonal antibodies and enable the detection of triazines with high binding affinity. The presented prototype is the first parallelized optical label-free detection system for biomolecular interaction analysis that is suitable for a high-throughput screening based on the 96-well microplate format. PMID- 11866065 TI - Conjugation of luminescent quantum dots with antibodies using an engineered adaptor protein to provide new reagents for fluoroimmunoassays. AB - We describe the preparation and characterization of bioinorganic conjugates made with highly luminescent semiconductor CdSe-ZnS core-shell quantum dots (QDs) and antibodies for use in fluoroimmunoassays. The conjugation strategy employs an engineered molecular adaptor protein, attached to the QDs via electrostatic/hydrophobic self-assembly, to link the inorganic fluorophore with antibodies. In this method, the number of antibodies conjugated to a single QD can be varied. In addition, we have developed a simple purification strategy based on mixed-composition conjugates of the molecular adaptor and a second two domain protein that allows the use of affinity chromatography. QD-antibody conjugates were successfully used in fluoroimmunoassays for detection of both a protein toxin (staphylococcal enterotoxin B) and a small molecule (2,4,6 trinitrotoluene). PMID- 11866066 TI - Size characterization of incinerator fly ash using sedimentation/steric field flow fractionation. AB - Fly ash particles emitted from municipal solid waste-incinerators are of environmental concern. This study aims to investigate the applicability of sedimentation/steric field-flow fractionation (Sd/StFFF) and to develop a Sd/StFFF method for the separation and size characterization of incinerator fly ash. This study focuses on the fly ash particles larger than approxiamtely 1 microm, which comprise more than 90% (w/w) of the fly ash. Fly ash is a complex mixture of particles having various chemical compositions, sizes, shapes, and densities. Prior to Sd/StFFF analysis, fly ash particles are prefractionated into six density classes using a modified centrifugal procedure. It was found that fly ash particles are most abundant in the density range between 2.4 and 2.8 g/cm3. Different density fractions seem to contain particles of different chemical compositions. The Sd/StFFF conditions for the size-characterization of fly ash are sample concentration, approximately 0.3% (w/v); dispersing medium, 50% ethanol in water; and carrier liquid, water with 1.0% FL-70 (ionic strength approximately 0.012 M). Sd/StFFF data show no significant differences in size distribution among different density fractions. Generally, the sizes obtained from Sd/StFFF are larger than those obtained from a Coulter Multisizer and microscopy, probably because of the irregular shapes of the fly ash particles. PMID- 11866067 TI - Surface characterization of laser-ablated polymers used for microfluidics. AB - Fabrication of microfluidic devices by excimer laser ablation under different atmospheres may provide variations in polymer microchannel surface characteristics. The surface chemistry and electroosmotic (EO) mobility of polymer microchannels laser ablated under different atmospheres were studied by X ray photoelectron spectroscopy and current monitoring mobility measurements, respectively. The ablated surfaces of PMMA were very similar to the native material, regardless of ablation atmospheres due to the negligible absorption of 248-nm light by that polymer. The substrates studied that exhibit nonnegligible absorption at this energy, namely, poly(ethylene terephthalate glycol), poly(vinyl chloride), and poly(carbonate), showed significant changes in surface chemistry and EO mobility when the ablation atmospheres were varied. Ablation of these three polymer substrates under nitrogen or argon resulted in low EO mobilities with a loss of the well-defined chemical structures of the native surfaces, while ablation under oxygen yielded surfaces that retained native chemical structures and supported higher EO mobilities. PMID- 11866068 TI - Scanning tunneling microscopic and voltammetric studies of the surface structures of an electrochemically activated glassy carbon electrode. AB - The microscopic surface structures of electrochemically activated glassy carbon have been examined by scanning tunneling microscopy. Experimental results demonstrate that there are two different types of electrode surface sites, corresponding to the bundles and bundle edges of the fibrous graphite microcrystallities. Electrochemical activation results in the formation of new void spaces of different sizes, and the structures are affected by the electrochemical activation procedures employed. The void volume resulting from cyclic polarization is usually smaller than that obtained by potentostatic activation. Electrochemical behaviors of the activated electrode are related to both the new void structures and the size of the electroactive species employed. On the basis of the STM voltammetric results, the microscopic structure of the activated electrode is proposed. PMID- 11866069 TI - Aluminum surface corrosion and the mechanism of inhibitors using pH and metal ion selective imaging fiber bundles. AB - The localized corrosion behavior of a galvanic aluminum copper couple was investigated by in situ fluorescence imaging with a fiber-optic imaging sensor. Three different, but complementary methods were used for visualizing remote corrosion sites, mapping the topography of the metal surface, and measuring local chemical concentrations of H+, OH-, and Al3+. The first method is based on a pH sensitive imaging fiber, where the fluorescent dye SNAFL was covalently attached to the fiber's distal end. Fluorescence images were acquired as a function of time at different areas of the galvanic couple. In a second method, the fluorescent dye morin was immobilized on the fiber-optic imaging sensor, which allowed the in situ localization of corrosion processes on pure aluminum to be visualized over time by monitoring the release of Al3+. The development of fluorescence on the aluminum surface defined the areas associated with the anodic dissolution of aluminum. We also investigated the inhibition of corrosion of pure aluminum by CeCl3 and 8-hydroxyquinoline. The decrease in current, the decrease in the number of active sites on the aluminum surface, and the faster surface passivation are all consistent indications that cerium chloride and 8 hydroxyquinoline inhibit corrosion effectively. From the number and extent of corrosion sites and the release of aluminum ions monitored with the fiber, it was shown that 8-hydroxyquinoline is a more effective inhibitor than cerium chloride. PMID- 11866070 TI - Electrochemical oxidation of chlorophenols at a boron-doped diamond electrode and their determination by high-performance liquid chromatography with amperometric detection. AB - Anodically pretreated diamond electrodes have been used for the detection of chlorophenols (CPs) in environmental water samples after high-performance liquid chromatographic (HPLC) separation. The anodization of as-deposited boron-doped polycrystalline diamond thin-film electrodes has enabled the stable determination of phenols over a wide concentration range. Prior to the HPLC analysis, a comparative study with ordinary glassy carbon, as-deposited diamond, and anodized diamond was made to examine the oxidative behavior of phenols by cyclic voltammety and flow injection analysis with amperometric detection. At anodized diamond electrodes, reproducible, well-defined cyclic voltammograms were obtained even at high CP concentration (5 mM), due to a low proclivity for adsorption of the oxidation products on the surface. In addition, after prolonged use, the partially deactivated diamond could be reactivated on line by applying a highly anodic potential (2.64 Vvs SCE) for 4 min, which enabled the destruction of the electrodeposited polymer deposits. Hydroxyl radicals produced by the high applied potential, in which oxygen evolution occurs, are believed to be responsible for the oxidation of the passivating layer on the surface. When coupled with flow injection analysis (FIA), anodized diamond exhibited excellent stability, with a response variability of 2.3% (n = 100), for the oxidation of a high concentration (5 mM) of chlorophenol. In contrast, glassy carbon exhibited a response variability of 39.1%. After 100 injections, the relative peak intensity, for diamond decreased by 10%, while a drastic decrease of 70% was observed for glassy carbon. The detection limit obtained in the FIA mode for 2,4-dichlorophenol was found to be 20 nM (S/N = 3), with a linear dynamic range up to 100 microM. By coupling with the column-switching technique, which enabled on-line preconcentration (50 times), the detection limit was lowered to 0.4 nM (S/N = 3). By use of this technique, anodized diamond electrodes were demonstrated for the analysis of CPs in drainwater that was condensed from the flue gas of waste incinerators. PMID- 11866071 TI - Amperometric quantification of total coliforms and specific detection of Escherichia coli. AB - The quantitative determination of total and fecal coliforms, as indicators of fecal pollution, is essential for water quality control. We developed a sensitive, inexpensive amperometric enzyme biosensor based on the electrochemical detection of beta-galactosidase activity, using p-amino-phenyl-beta-D galactopyranoside as substrate, for determining the density of coliforms, represented by Escherichia coli and Klebsiella pneumoniae. The specific detection of E. coli was achieved using an antibody-coated electrode that specifically binds the target bacteria. Amperometric detection enabled the determination of 1000 colony-forming units/mL within 60-75 min. Preincubation for 5-6 h further increased the sensitivity more than 100-fold. The present experimental setup allowed the simultaneous analysis of up to eight samples, using disposable screen printed electrodes. PMID- 11866072 TI - Ultra-low-volume, real-time measurements of lactate from the single heart cell using microsystems technology. AB - The fabrication of microelectrodes integrated within ultra-low-volume microtiter chambers for the amperometric determination of metabolites continues to be of interest in the subject of single-cell and high-throughput screening. The microsystem described in this paper consists of a two-microelectrode sensor with a microfluidic dispensation technology, which is able to deliver both very low titers (6.5 pL) and single heart cells into a low-volume microphotoelectrochemical cell. Devices were fabricated using photolithography and liftoff giving reproducible sensors integrated within high aspect ratio titer chambers (with a volume of 360 pL), made of the photoepoxy SU8. In this paper, the determination of lactate was optimized using an enzyme-linked assay based upon lactate oxidase, involving the amperometric determination of hydrogen peroxide at +640 mV versus an internal Ag/AgCl pseudoreference. The microsystem (including the microfluidic dispensers and structures as well as the microsensor) was subsequently used to measure the lactate content of single heart cells. Dynamic electrochemical measurements of lactate during cell permeabilization are presented. We also show the use of respiratory uncouplers to simulate ischemia in the single myocyte and show that, as expected, the rate of lactate production from the hypoxic heart cell is greater than that within the normoxic healthy myocyte. PMID- 11866073 TI - Flow-through cell for on-line amperometric determination of Ce(lV) during polymerization reactions. AB - In this paper, a method is described for the chronoamperometric determination of Ce(IV) in acidic aqueous solutions in order to study the kinetics and the mechanism of polymerization reactions with Ce(IV) as the initiator. The method is based on the FIA principle, in which small samples are injected in a continuous 1.0 mol L(-1) H2SO4 flow, and Ce(IV) is detected chronoamperometrically by reduction at the surface of a gold CDtrode. Such an electrode is made from a commercial CD that has a gold coating acting as a reflecting layer for the laser beam. It was found that a detection limit of 1.5 10(-7) mol L(-1) could be obtained with this method, that high concentrations (order of 10(-2) mol L(-1)) can be detected without IR drop and that the system can be used over several days without renewal or recalibration of the gold CDtrode. PMID- 11866074 TI - A method for screening total mercury in water using a flow injection system with piezoelectric detection. AB - An automatic microgravimetric screening system based on piezoelectric detection and the use of acidic stannous chloride as reductant was developed for the fast detection and determination of total mercury in water. Reduced mercury is detected as an amalgam by using a gold-coated piezoelectric crystal, the sensor subsequently being regenerated by passing it through a peroxydisulfate solution. The gold-coated piezoelectric crystal is a highly efficient retention unit for the main soluble mercury species (inorganic, complexed, and organometallic) previously reduced to elemental mercury and is free of interferences from other metal ions. This detector exhibits good sensitivity: it allows the determination of mercury at sub-parts-per-billion concentration levels (0.30-1.00 microg/L). The precision, expressed as relative standard deviation, was +/- 2.7% (n = 11; P = 0.05) at 0.5 microg/L total mercury. The proposed method was successfully used as a rapid screening method for mercury monitoring in natural waters. PMID- 11866076 TI - Interobserver reliability in the interpretation of radiologic signs in Legg-Calve Perthes disease. AB - An accurate interpretation of radiographs is crucial in determining therapeutic choices in Legg-Calve-Perthes disease. The aim of this study was to measure the interobserver reliability of a group of surgeons in this condition. Twenty-three radiographs were shown to nine pediatric orthopedic surgeons at nine different medical centers. Differences in coding between members of the group was assessed using a two-way random factor analysis of variance. Reliability was excellent for the Catterall classification [intraclass correlation coefficient (ICC) = 0.94], and good for the stage of disease and Head at Risk signs (ICC ranging from 0.74 to 0.85) except for lateral subluxation, whose reliability is fair (ICC = 0.68). Analysis of results shows that lateral subluxation should be expressed in a quantitative measurement rather than a binary answer. The reliability of Gage's sign would be improved if multiple definitions did not exist in the literature. In experienced hands, Catterall's classification can be used with reliability. Difficulties are still noted in separating group II frorm group III. PMID- 11866075 TI - Infantile spinal cord tumor: diagnostic difficulties. A case report. AB - We report an infant with a thoracic intradural extramedullary tumor to illustrate the presentation of an infantile spinal cord tumor and the difficulty in establishing a diagnosis. An infant presented with edema of the legs, motor loss in the lower extremities, and anuria at 3 months of age. Magnetic resonance imaging of the lumbar region did not reveal any abnormalities. At 1 year of age, the patient underwent surgery to correct urinary incontinence of unknown etiology. The patient developed gait disturbance and had abdominal pain at 17 months of age. Magnetic resonance imaging of the thoracic spine revealed an intradural extramedullary tumor at T2-T5. The spinal cord tumor was completely resected and pathologic findings indicated an endodermal cyst. All symptoms improved after the operation. Infants who present with weakness of the lower legs, urinary incontinence and unknown abdominal pain should be evaluated for a spinal cord tumor. PMID- 11866077 TI - Comparison between Salter's innominate osteotomy and augmented acetabuloplasty in the treatment of patients with severe Legg-Calve-Perthes disease. Analysis of 90 hips with special reference to roentgenographic sphericity and coverage of the femoral head. AB - This is a roentgenographic and retrospective study comparing the results of two different operative procedures used in the treatment of Legg-Calve-Perthes disease: Salter innominate osteotomy, and Augmented Acetabuloplasty. Group I comprised 43 patients (50 hips) who underwent Salter osteotomy. Another 40 patients (40 hips) were treated by Augmented Acetabuloplasty and formed Group 2. In Group 1, 11 (25.58%) patients were girls and 32 (74.42%) were boys. In Group 2, there were 4 (10%) girl and 36 (90%) boy patients. The average age was 6.62 years in Group 1 and 6.35 years in Group 2. Follow-up varied from 2 years to 10 years in both Groups 1 and 2. In Group 1, 18 (36%) hips were in the stage of necrosis, 28 (56%) in fragmentation, and 4 (8%) in reossification; in Group 2, 16 (40%) hips were in necrosis, 23 (57.5%) in fragmentation, and only 1 (2.5%) in reossification. All hips in necrosis were included in Salter and Thompson group B; all hips in fragmentation were classed as Catterall group 3 or 4. Preoperative arthrographs of the 50 hips in Group I were graded according to Laredo: 29 (58%) in group III, 18 (36%) in group IV, and 3 (6%) in group V. Only 23 of the 43 hips of Group 2 had preoperative arthrographs; there were 14 (60.87%) in group III, 8 (34.78%) in group IV, and 1 (4.35%) in group V. The preoperative Wiberg's Center Edge (CE) angle of Group 2 (19.4 degrees) was significantly greater than that of Group 1 (16.7 degrees). The percentage difference (A%) between immediate postoperative and preoperative CE angle was significantly greater in Group 2. The delta% between final and immediate postoperative CE angle was significantly greater in Group 1, and within Group 1 it was significantly greater at age 4 years to 6 years. The A% between final and preoperative CE angle was statistically the same in both Group I and Group 2. The distribution of good, fair and poor Mose ratings did not differ in Group 1 and Group 2, but there was a tendency of better results after Augmented Acetabuloplasty in children older than 6 years. Younger patients of Group 1 (4-6 years) ended up with good results at a significantly higher frequency than the older ones. Group 1 children with preoperative Laredo III arthrography had good results in a significantly greater number of cases, as compared with Laredo grade IV. The delta% between final and immediate postoperative CE angle was greater in those hips with good Mose rating. PMID- 11866078 TI - Seasonal variation of slipped capital femoral epiphysis. AB - During the 18-year period 1980-1997, 1103 patients were treated as in-patients for slipped capital femoral epiphysis (SCFE) in Scottish hospitals. This paper reports a significant seasonal variation, especially in male patients, with an excess admitted in the autumn. Examining the physiological seasonality of the monthly increment of growth in height and weight in childhood, we hypothesised that these physiological rhythms, while not pathogenetic, may be responsible for the timing of the seasonal features of SCFE. Previously published studies suggest that the average time between first symptoms and diagnosis is 3 to 5 months. The condition may start with the spring peak in growth and become more symptomatic with the autumn peak in weight. In conclusion, we report a new epidemiological finding, but, in accordance with other studies, we cannot provide a certain aetiological explanation. The timing, but not the pathogenesis, of admission to hospital for a SCFE might be related to the timing in the year of seasonal increments of height in spring and weight in autumn. It is tempting to draw attention to associations with environmental features. For example, in autumn there is the most rapid annual fall in temperature, and the hours of darkness are increasing from the autumn equinox to the winter solstice. However, there is no reason to conclude that these associations have anything to do with aetiology. PMID- 11866080 TI - Evaluation of the risk factors of avascular necrosis of the femoral head in developmental dysplasia of the hip in infants younger than 18 months of age. AB - Sixty-seven dysplastic hips of 54 infants with an average age of 11.9 months were treated by the same surgical technique, including iliopsoas and adductor tenotomies via medial approach and arthrography for verification of the concentric reduction. The average follow-up period was 44.5 months. The rate of overall avascular necrosis (AVN) was 27% and that of severe AVN was 7%. With the numbers available in the study, a statistically significant relationship could not be found between the rate and severity of AVN and several preoperative and intraoperative components such as age, sex, side, dislocation grade, development of the ossific nucleus and qualitv of intraoperative reduction. Nevertheless, hips of infants treated between 13 months and 15 months of age, hips of male patients and left hips had slightly higher ratios of ischemic changes that were not statistically significant. PMID- 11866079 TI - Retroversion of the acetabular dome after Salter and triple pelvic osteotomy for congenital dislocation of the hip. AB - Retroversion of the acetabular dome has been associated with hip pain and osteoarthritis in several studies. Nevertheless, this acetabular alignment received little attention when studying the radiological outcome of pelvic osteotomies in childhood. We therefore reviewed the charts and X-rays of 73 patients with congenital dislocations, who underwent 97 pelvic osteotomies in childhood. We focussed our attention on anterior overcoverage or retroversion of the acetabular dome, respectively. Eighty-six Salter and 11 LeCoeur osteotomies were performed on patients with a mean age of 4.8 years. The mean age at last X ray documentation was 16.5 years. The version of the acetabular dome was estimated qualitatively from the relationship of the anterior and posterior border to each other and measured semiquantitatively using templates developed by Hefti. According to Lequesne's criteria 94% of the hips were normal or borderline at maturity. A retroverted acetabular dome was present in 27% and averaged -15 degrees. It was more frequent (60% versus 24%) and more pronounced (-16 degrees versus +5 degrees) after Le Coeur's than after Salter's osteotomy and in the residually dysplastic hips (83% versus 22%; -15 degrees versus +3 degrees). It is suggested that anterior overcoverage of the femoral head results from ignorance, when choosing the type and performing the pelvic osteotomy, of where coverage needs to be improved in an individual hip. We question the long-term outcome of these hips because such rotational misalignment has been suggested to be associated with early hip pain and osteoarthritis. In order to avoid such misalignment, preoperative evaluation of where coverage needs to be improved in an individual hip and efforts for better intraoperative control of acetabular reorientation should be made. PMID- 11866082 TI - A new knee arthroplasty versus Brown procedure in congenital total absence of the tibia: a preliminary report. AB - The rates of successful knee arthroplasties according, for example, to Brown in patients with congenital total absence of the tibia are disappointing. A new form of knee arthroplasty is demonstrated in a 15-month-old infant with a congenital total absence of the tibia. With the help of two crossed capsular flaps and a Z plasty of the quadriceps tendon, the patella is transposed under the femoral condyles to function as a tibial plateau. The proximal fibula is fused centrally to the external, cortical surface of the patella. A mini-ringfixator construction is used to stabilize the components, although permitting mobilization of the knee arthroplasty in the immediate postoperative period. This new type of knee arthroplasty offers distinct advantages compared with the previously published procedures. PMID- 11866081 TI - Neurovascular calcaneo-cutaneus pedicle graft for stump capping in congenital pseudarthrosis of the tibia: preliminary report of a new technique. AB - The operative treatment of congenital pseudarthrosis of the tibia, especially when associated with neurofibromatosis type I (Recklinghausen's disease), often leads to failure. Frequently, regardless of the type of deformity, multiple operative procedures end in the amputation of the affected limb. Soft tissue coverage of the amputation stump may confront the surgeon with new problems. Secondary perforation of the soft tissue envelope of the stump owing to terminal overgrowth is not a rare complication. A new technique of stump capping is demonstrated in a 10-year-old boy and a 14-year old girl, both with congenital pseudarthrosis of the tibia of the right leg and neurofibromatosis. During this procedure, a radical resection of the pseudarthrotic tissue is performed although all neurovascular structures supplying the calcaneus are carefully maintained (Arteria, vena et nervus tibialis posterior et peronealis). Subsequently, the tibia and fibula are inserted into the calcaneus. This construct is stabilized with two crossed Kirschner wires. Afterwards, the skin of the lower limb and the hindfoot are sutured together covering the neurovascular bundles, which are arranged in a loop-like fashion. The documented active range of motion was similar to that of the unaffected knee joint. Twelve weeks after operation in both patients, full weight bearing was achieved with a lower-leg prosthesis. This new procedure leads to a durable, full weight-bearing stump with complete sensitive innervation without the risk of future soft tissue perforation caused by the growing bone. The stump fits with an end-bearing lower-leg prosthesis. PMID- 11866083 TI - Permanent thoracic myelopathy resulting from herniation of a calcified intervertebral disc in a child. AB - This retrospective study was made to illustrate the rare occurrence of neurologic deficits resulting from intervertebral disc calcification (IDC) in a child. Most authors agree that juvenile IDC is usually a benign, self-limiting disease with excellent prognosis. The symptoms subside spontaneously in 95% of patients. Conservative treatment is therefore usually sufficient. Reviewing the English speaking literature, only two further cases of operated juvenile IDC with myelopathy have been published. In the current report, we describe a case of permanent thoracic myelopathy resulting from juvenile IDC treated by urgent decompressive thoracic laminectomy. At the 3-year follow-up examination, the patient had not recovered fully. Persisting deficits in motor and sensory function were observed. PMID- 11866084 TI - Single stage surgical correction of congenital vertical talus by complete subtalar release and peritalar reduction by using the Cincinnati incision. AB - Seventeen feet of 12 patients with congenital vertical talus, which were treated with a single-stage surgical correction of complete subtalar release and peritalar reduction by using the Cincinnati incision, were reviewed. Five of the feet were associated with neuromuscular disorders, four with neural tube defects, and eight were idiopathic. All patients were operated on by the same surgeon and all were available for clinical and radiographic follow-up averaging 42.9 months (range 19-81) from the time of surgery. There were no wound complications or avascular necrosis of the talus. None required subsequent reoperation. At final follow-up, results were based on clinical and radiographic outcomes and included five excellent, eight good, two fair, and two poor. All families were satisfied with the results and appearance of the feet. Radiographically, there was a significant improvement in the anteroposterior and lateral talocalcaneal and talo first metatarsal angles, and at follow-up, the group averages for each of these angles were within the normal ranges. In the treatment of congenital vertical talus, good clinical and radiographic outcomes can be obtained, with a low incidence of complications, using this single-stage surgical correction procedure. PMID- 11866085 TI - Foot lengthening using the Ilizarov device: the transverse tarsal joint resection versus osteotomy. AB - The aim of the study was the evaluation of both the foot correction and foot lengthening obtained using the distraction method with osteotomies versus distraction after the transverse tarsal joint resection. Ten patients (10 feet) aged from 5 years to 24 years (average, 10.5 years) were analyzed. Seven of them were treated for severe equinovarus deformity: six of congenital and one of post traumatic etiology. In three patients, the indication for treatment was foot shortening due to hypoplasia with tibial shortening, combined with foot deformity. Preoperative shortening of the foot ranged from 1.5 cm to 10 cm (average, 4.5 cm). In four patients, osteotomy between the tarsometatarsal and transverse tarsal joints was carried out. In two cases, 'V-shaped' osteotomy through the hindfoot and midfoot was performed. In the remaining four patients, wedge resection of the transverse tarsal joint was performed. The follow-up was a mean of 32 months (range, 12-55 months). It was observed that foot lengthening after transverse osteotomy of the midfoot is difficult and unpredictable, because of distraction at the adjacent joints level instead of osteotomy site. The greatest lengthening of the foot (mean, 4 cm) was observed in the patients with the transverse tarsal joint resection. It was concluded that the transverse tarsal joint resection following callus distraction in the place of the resected joint is the effective method for foot lengthening, which can be combined with deformity correction. PMID- 11866087 TI - Pre-axial polydactyly: outcome of the surgical treatment. AB - From 1988 to 1999 we operated on 22 patients for pre-axial polydactyly. We retrospectively reviewed all files and we clinically reviewed 16 patients. We report on the epidemiology, the surgical technique, the outcome and the complications. All patients were operated on by the same hand surgeon (L.D.S.) and reviewed by an independent observer (S.K.). Aesthetic and functional outcome is reported. In general the surgical treatment for pre-axial duplications gives excellent functional and good aesthetic results. According to the Egawa score, 13 patients obtained an excellent, one a fair and two a poor result. The overall satisfaction rate is very high for both patient and parents and the complication rate was low. The functional outcome seems to be related to the stability of the interphalangeal and metacarpo-phalangeal joints. The most frequent aesthetic problem is a deviation of the axis. Loss of mobility of the thumb seems to be of less importance. PMID- 11866086 TI - Avascular necrosis of the talus in children with haemophilia. AB - Patients who have haemophilia often present with joint problems. Haemarthrosis can occur spontaneously or after minor trauma. Later, degenerative changes can cause pain and loss of motion. Avascular necrosis of the talus has a high incidence after serious trauma. Furthermore, avascular necrosis of the talus has been associated with multiple systemic conditions. The association of haemophilia and avascular necrosis of the femoral head has been described. This study reports three cases (four ankles) where we suspect an avascular necrosis of the talus in children with haemophilia. To the authors' knowledge, the possible association of haemophilia and avascular necrosis of the talus has not yet been described in children. PMID- 11866088 TI - Congenital high scapula. AB - Congenital high scapula is also known as Sprengel's deformity. The elevation of the scapula is accompanied by its rotation to a varus position. A series of 19 cases is presented, with 4 bilateral cases. A modified Woodward procedure was performed in all the surgical cases using the basic Woodward technique modified by correcting the tilting of the glena. The operative results were judged on cosmetic and functional criteria. The age of the patients and the presence of an omovertebral bone did not influence the results. Associated cervical spine anomalies were of negative prognosis. Results in this series showed only three fair or poor results; the other cases (79%) were all improved cosmetically and functionally with normal shoulder abduction following reorientation of the scapulo-humeral joint. PMID- 11866089 TI - Growth arrest of the distal radius following a metaphyseal fracture: case report and review of the literature. AB - We report a 12-year-old girl who developed growth arrest of the distal radius physis 9 months after sustaining a complete fracture of the distal radial and ulnar metaphysis with no involvement of the physis evident at time of injury. The girl sustained a fracture of the metaphysis of her right distal radius and ulna after a fall. Anterior-posterior, lateral and oblique radiographs at injury, and during subsequent healing show no evidence of the fracture involving the physis. She was treated with closed reduction and casting for 6 weeks and healed uneventfully. She returned 4 month later concerned about distal ulnar prominence. Radiographs revealed a loss of radial tilt and with suspicion of a physeal bar. Magnetic resonance imaging confirmed a physeal bar located in the dorsal radial region. A literature search of the Medline database was used to obtain prior case reports for review purpose. The patient underwent an epiphysiodesis of the distal radius and ulna along with an opening wedge osteotomy and bone grafting of the distal radius to restore radial height and inclination. She healed without complication and with restoration of the normal relationship of the distal radius and ulna. A review of the literature reveals five reported case of distal radial metaphyseal fractures not invloving the physis leading to growth arrest. By comparison, there are 31 reported cases of distal radius physeal arrest following fractures involving the physis. The physician should be aware that common distal radius metaphyseal fractures may rarely lead to growth arrest. PMID- 11866090 TI - Moonlighting functions of polypeptide elongation factor 1: from actin bundling to zinc finger protein R1-associated nuclear localization. AB - Eukaryotic polypeptide elongation factor EF-1 is not only a major translational factor, but also one of the most important multifunctional (moonlighting) proteins. EF-1 consists of four different subunits collectively termed EF 1alphabeta beta'gamma and EF-1alphabeta gammadelta in plants and animals, respectively. EF-1alpha x GTP catalyzes the binding of aminoacyl-tRNA to the A site of the ribosome. EF-1beta beta'gamma (EF-1beta and EF-1beta'), catalyzes GDP/GTP exchange on EF-1alpha x GDP to regenerate EF-1alpha x GTP. EF-1gamma has recently been shown to have glutathione S-transferase activity. EF-2 catalyzes the translocation of peptidyl-tRNA from the A-site to the P-site on the ribosome. Recently, molecular mimicry among tRNA, elongation factors, releasing factor (RF), and ribosome recycling factor (RRF) has been demonstrated and greatly improved our understanding of the mechanism of translation. Moreover, eukaryotic elongation factors have been shown to be concerned or likely to be concerned in various important cellular processes or serious diseases, including translational control, signal transduction, cytoskeletal organization, apoptosis, adult atopic dermatitis, oncogenic transformation, nutrition, and nuclear processes such as RNA synthesis and mitosis. This article aims to overview the recent advances in protein biosynthesis, concentrating on the moonlighting functions of EF-1. PMID- 11866091 TI - On the interaction of ribosomal protein L5 with 5S rRNA. AB - Ribosomal protein L5, a 5S rRNA binding protein in the large subunit, is composed of a five-stranded antiparallel beta-sheet and four alpha-helices, and folds in a way that is topologically similar to the ribonucleprotein (RNP) domain [Nakashima et al., RNA 7, 692-701, 20011. The crystal structure of ribosomal protein L5 (BstL5) from Bacillus stearothermophilus suggests that a concave surface formed by an anti-parallel beta-sheet and long loop structures are strongly involved in 5S rRNA binding. To identify amino acid residues responsible for 5S rRNA binding, we made use of Ala-scanning mutagenesis of evolutionarily conserved amino acids occurred at beta-strands and loop structures in BstL5. The mutation of Lys33 at the beta 1-strand caused a significant reduction in 5S rRNA binding. In addition, the Arg92, Phe122, and Glu134 mutations on the beta2-strand, the alpha3-beta4 loop, and the beta4-beta5 loop, respectively, resulted in a moderate decrease in the 5S rRNA binding affinity. In contrast, mutation of the conserved residue Pro65 at the beta2-strand had little effect on the 5S rRNA binding activity. These results, taken together with previous results, identified Lys33, Asn37, Gln63, and Thr90 on the beta-sheet structure, and Phe77 at the beta2-beta3 loop as critical residues for the 5S rRNA binding. The contribution of these amino acids to 5S rRNA binding was further quantitatively evaluated by surface plasmon resonance (SPR) analysis by the use of BIAcore. The results showed that the amino acids on the beta-sheet structure are required to decrease the dissociation rate constant for the BstL5-5S rRNA complex, while those on the loops are to increase the association rate constant for the BstL5-5S rRNA interaction. PMID- 11866092 TI - Effects of amino acid alterations on the transglycosylation reaction of endoglucanase I from Trichoderma viride HK-75. AB - Endoglucanase I (EGI) from Trichoderma viride HK-75 catalyzes not only hydrolysis but also transglycosylation reactions of cellooligosaccharides. In order to characterize the important amino acid residues in transglycosylation of EGI, three Tyr, one Leu, and two Glu residues of EGI were replaced by Trp or Asp. The seven resulting EGI, except for L200W, had reduced activities toward carboxymethyl-cellulose compared to that of wild type EGI. The results from the mutations in the catalytic residues of E196 and E201 indicate that the space just around the catalytic residues is not directly related to the transglycosylation reactions of EGI. Analyses of the enzymes with mutations in the substrate-binding residues showed that Y146, Y170, and L200 of EGI are closely involved in the mode of transglycosylation and that several amino acid residues within the active site are involved in the transglycosylation reaction of EGI. PMID- 11866093 TI - Effects of protein deprivation on alpha1(I) and alpha1(III) collagen and its degrading system in rat skin. AB - Protein malnutrition affects the status of dermal collagen, the major structural protein in the skin. However, the molecular mechanism underlying the alteration of collagen fibers in the skin by protein deficiency remains unknown. In the present study, the effect of dietary protein deprivation on collagen metabolism was studied by analyzing the status of the synthesis and degradation of collagen in the dorsal skin of rats. Feeding on a protein-free diet for 8 days caused a dramatic decrease in both types I and III tropocollagen with a concomitant decrease in their mRNA levels, with type III collagen being more severely affected. The active form of collagenase was significantly decreased by protein deprivation, whereas the latent form was not affected. The mRNA levels of collagenase and its inhibitors (TIMP-1 and 2) were also decreased by protein deprivation. These results suggest that both the synthesis and degradation of types I and III collagen were affected by protein deficiency. PMID- 11866094 TI - Substrate specificity at the P1' site of Escherichia coli OmpT under denaturing conditions. AB - Though OmpT has been reported to mainly cleave the peptide bond between consecutive basic amino acids, we identified more precise substrate specificity by using a series of modified substrates, termed PRX fusion proteins, consisting of 184 residues. The cleavage site of the substrate PRR was Arg140-Arg141 and the modified substrates PRX substituted all 19 natural amino acids at the P1' site instead of Arg141. OmpT under denaturing conditions (in the presence of 4 M urea) cleaved not only between two consecutive basic amino acids but also at the carboxyl side of Arg140 except for the Arg140-Asp141, -Glu141, and -Pro141 pairs. In addition to Arg140 at the P1 site, similar results were obtained when Lys140 was substituted into the P1 site. In the absence of urea, an aspartic acid residue at the P1' site was unfavorable for OmpT cleavage of synthetic decapeptides, the enzyme showed a preference for a dibasic site. PMID- 11866095 TI - 3-(4-Methyl-3-pentenyl)-2(5H)-furanone, alpha,alpha-acariolide and 4-(4-methyl-3 pentenyl)-2(5H)-furanone, alpha,beta-acariolide: new monoterpene lactones from the astigmatid mites, Schwiebea araujoae and Rhizoglyphus sp. (Astigmata: Acaridae). AB - A new monoterpene lactone from the acarid mite, Schwiebea araujoae, was elucidated without its isolation by GC/FT-IR and GC/MS analyses to be 3-(4-methyl 3-pentenyl)-2(5H)-furanone (1) and tentatively named as alpha,alpha-acariolide. The structure of 1 was identified by its synthesis from alpha-bromo-gamma butyrolactone via 4 reaction steps. The synthesized compound gave the same GC/MS and GC/FT-IR spectra as those of the natural product. The other monoterpene lactone was likewise elucidated from the unidentified Rhizoglyphus mite to be 4 (4-methyl-3-pentenyl)-2(5H)-furanone (2) and named as alpha,beta-acariolide; it was also identified by its synthesis in 5 reaction steps from the same butyrolactone as the starting material. GC/MS and GC/FT-IR spectra of the preparation were identical to those of the natural product. PMID- 11866097 TI - The primary structure of cassowary (Casuarius casuarius) goose type lysozyme. AB - The complete amino acid sequence of cassowary (Casuarius casuarius) goose type lysozyme was analyzed by direct protein sequencing of peptides obtained by cleavage with trypsin, V8 protease, chymotrypsin, lysyl endopeptidase, and cyanogen bromide. The N-terminal residue of the enzyme was deduced to be a pyroglutamate group by analysis with a LC/MS/MS system equipped with the oMALDI ionization source, and then confirmed by a glutamate aminopeptidase enzyme. The blocked N-terminal is the first reported in this enzyme group. The positions of disulfide bonds in this enzyme were chemically identified as Cys4-Cys60 and Cys18 Cys29. Cassowary lysozyme was proved to consist of 185 amino acid residues and had a molecular mass of 20408 Da calculated from the amino acid sequence. The amino acid sequence of cassowary lysozyme compared to that of reported G-type lysozymes had identities of 90%, 83%, and 81%, for ostrich, goose, and black swan lysozymes, respectively. The amino acid substitutions at PyroGlu1, Glu19, Gly40, Asp82, Thr102, Thr156, and Asn167 were newly detected in this enzyme group. The substituted amino acids that might contribute to substrate binding were found at subsite B (Asn122Ser, Phe123Met). The amino acid sequences that formed three alpha-helices and three beta-sheets were completely conserved. The disulfide bond locations and catalytic amino acid were also strictly conserved. The conservation of the three alpha-helices structures and the location of disulfide bonds were considered to be important for the formation of the hydrophobic core structure of the catalytic site and for maintaining a similar three-dimensional structure in this enzyme group. PMID- 11866098 TI - Amino acid sequence and carbohydrate-binding analysis of the N-acetyl-D galactosamine-specific C-type lectin, CEL-I, from the Holothuroidea, Cucumaria echinata. AB - CEL-I is one of the Ca2+-dependent lectins that has been isolated from the sea cucumber, Cucumaria echinata. This protein is composed of two identical subunits held by a single disulfide bond. The complete amino acid sequence of CEL-I was determined by sequencing the peptides produced by proteolytic fragmentation of S pyridylethylated CEL-I. A subunit of CEL-I is composed of 140 amino acid residues. Two intrachain (Cys3-Cys14 and Cys31-Cys135) and one interchain (Cys36) disulfide bonds were also identified from an analysis of the cystine-containing peptides obtained from the intact protein. The similarity between the sequence of CEL-I and that of other C-type lectins was low, while the C-terminal region, including the putative Ca2+ and carbohydrate-binding sites, was relatively well conserved. When the carbohydrate-binding activity was examined by a solid-phase microplate assay, CEL-I showed much higher affinity for N-acetyl-D-galactosamine than for other galactose-related carbohydrates. The association constant of CEL-I for p-nitrophenyl N-acetyl-beta-D-galactosaminide (NP-GalNAc) was determined to be 2.3 x 10(4) M(-1), and the maximum number of bound NP-GalNAc was estimated to be 1.6 by an equilibrium dialysis experiment. PMID- 11866096 TI - Differential effect of walnut oil and safflower oil on the serum cholesterol level and lesion area in the aortic root of apolipoprotein E-deficient mice. AB - Walnut oil (WO) is a good source of alpha-linolenic acid. We compared the effects of WO and high-linoleic safflower oil (HLSO) on the serum lipid level and atherosclerosis development in male and female apolipoprotein (apo) E-deficient mice. The WO diet resulted in a higher level of serum cholesterol than with HLSO. Female mice fed on the WO diet had a greater lesion area in the aortic root than did those on the HLSO diet. There was no diet-dependent difference in the level of cholesterol and its oxidation products in the abdominal and thoracic aorta. These results suggest that the unpleasant effects of the WO diet on apo E deficient mice may be attributable to alpha-linolenic acid. PMID- 11866099 TI - Purification and some properties of a keratinolytic enzyme from an alkaliphilic Nocardiopsis sp. TOA-1. AB - A novel alkaliphilic Nocardiopsis sp., strain TOA-1, was isolated from a tile joint of a bathroom. Strain TOA-1 produced a variety of alkaline hydrolytic enzymes. An alkaline protease, designated NAPase, was purified and characterized. NAPase had a very high keratinolytic activity and high stability under acidic conditions. PMID- 11866100 TI - Geraniol-inducible glutathione S-transferase in cultured soybean cells. AB - When the cultured cells of Glycine max (soybean) were treated with 5 mM geraniol as a chemical stress, an mRNA level was elevated in a rapid but transient increase. The mRNA was cloned and sequenced, and found to correspond to the mRNA encoding glutathione S-transferase (GST). The GST mRNA level and GST activity were elevated to maxima at 4-6 h and 8 h, respectively, after treatment of the cultures with geraniol. These indicate that GST is one of the geraniol-responsive factors in soybean cells. PMID- 11866101 TI - A variant of Orpinomyces joyonii 1,3-1,4-beta-glucanase with increased thermal stability obtained by random mutagenesis and screening. AB - A thermostable variant of an Orpinomyces joyonii beta-glucanase was identified by screening a mutant library constructed using error-prone PCR products. The mutant, designated 2011D, had one amino acid substitution (Val replaced Asp-70). 2011D showed similar catalytic efficiency to its wild-type enzyme, LicA. The temperature at which 50% inactivation occurred after heat treatment for 10 min was increased by 14 degrees C for 2011D, in comparison to those of wild-type enzyme. PMID- 11866102 TI - Inhibition of specific degradation of 57-kDa protein in royal jelly during storage by ethylenediaminetetraacetic acid. AB - We have previously shown that 57-kDa protein in royal jelly (RJ) was specifically degraded in proportion to both storage temperature and storage period, and we suggested that it could be useful as a marker of freshness of RJ (Kamakura, M., Fukuda, T., Fukushima, M. and Yonekura, M., Biosci. Biotechnol. Biochem., 65, 277 284 (2001).). Here, we investigated the effects of various proteinase inhibitors on proteinase activity in RJ and on the specific degradation of 57-kDa protein during storage. Ethylenediaminetetraacetic acid (EDTA), but not other inhibitors, inhibited the proteinase activity in RJ, and dose-dependently suppressed storage dependent degradation of 57-kDa protein. These results suggest that EDTA inhibits a specific proteinase activity in RJ, thereby suppressing the degradation of 57 kDa protein during storage at high temperature. PMID- 11866104 TI - Inhibition of translation and progesterone-induced maturation of Xenopus oocytes by expressing the amino-terminal portion of the eukaryotic translation initiation factor 4G. AB - The eukaryotic translation initiation factor 4G (eIF4G) plays a pivotal role in translation. EIF4G interacts with several other factors including eIF4E, which is a cap-binding protein, and the poly(A)-binding protein (PABP). In this work, we demonstrate that the expression of the amino-terminal one-third of eIF4G, which interacts with eIF4E and PABP, in Xenopus oocyte inhibits translation and progesterone-induced maturation. PMID- 11866103 TI - Anti-inflammatory compounds from the bitter mushroom, Sarcodon scabrosus. AB - A bioassay-guided purification procedure from the methanol extract of Sarcodon scabrosus led to the isolation of several anti-inflammatory compounds: sarcodonin A (1) and G (2), and related compounds (3, 4 and 5). We named these related compounds neosarcodonin A (3), B (4) and C (5) and elucidated their structures on the basis of spectral data. Topical application of each of these compounds to mouse ears suppressed TPA-induced inflammation. Neosarcodonin C (5) exhibited the highest activity and inhibited the TPA-induced edema on mouse ears by up to 87% with a 200-microg application. PMID- 11866105 TI - Isolation of yeast Kurtzmanomyces sp. I-11, novel producer of mannosylerythritol lipid. AB - Yeast strains were screened for producers of glycolipid-type biosurfactants from soybean oil as a sole carbon source. The structure of the glycolipid (MEL-I-11) produced by strain I-11 was analyzed. The hydrophilic sugar moiety was mannosylerythritol and the fatty acid components were C8:0 (36.4%), C12:0 (11.9%), and C14:2 (25.9%). The MEL-I-11 was identified as 6-O-acetyl-2,3-di-O alkanoyl-beta-D-mannopyranosyl-(1-->4)-O-meso-erythritol. The strain I-11 was identified as a Kurtzmanomyces species, a novel producer of mannosylerythritol lipid. PMID- 11866106 TI - Changes in N-acetyl-beta-D-glucosaminidase activity in the urine and urinary albumin excretion in magnesium deficient rats. AB - To discover the details of the effects of magnesium (Mg) deficiency on kidney function, the course of changes in N-acetyl-beta-D-glucosaminidase (NAG) activity in the urine and in urinary albumin excretion were examined in rats fed a Mg deficient diet. NAG activity in the urine and urinary albumin excretion in rats fed the Mg-deficient diet significantly increased from 7 d until the end of the feeding period. We suggest that Mg-deficient diet rapidly induces kidney function insufficiency. PMID- 11866107 TI - Uptake and physiological function of vitamin B12 in a photosynthetic unicellular coccolithophorid alga, Pleurochrysis carterae. AB - The photosynthetic coccolithophoid alga, Pleurochrysis (Hymenomonas) carterae, could take up and accumulate exogenous vitamin B12, most of which was converted into the coenzyme forms of vitamin B12. Two vitamin B12-dependent enzyme activities (methylmalonyl-CoA mutase, 2.6+/-0.4 nmol/min/mg protein and methionine synthase, 85.1+/-38.9 pmol/min/mg protein) could be found in a cell homogenate of the vitamin B12-supplemented alga. Most of the methylmalonyl-CoA mutase activity and 19.2% of the vitamin B12 accumulated by the algal cells were recovered in the macromolecular fractions with Mr of 150 kDa, although the remaining vitamin B12 was found only in free vitamin B12 fractions. PMID- 11866108 TI - Use of molecular-genetically bred Coprinus cinereus strains for an efficient isolation of cellulose from rice straw. AB - Molecular-bred Coprinus cinereus monokaryotic strains with high lignin- and xylan degrading activities were mixed-cultured at 27 degrees C in the liquid medium containing 0.5% (w/v) cut rice straw and 0.025% MnCl2. After 3 weeks, the culture supernatant was extensively treated with crude cellulase, showing the presence in it of 9.3% of the total cellulose of rice straw. When rice straw treated with 0.1 N NaOH or cultured with Ganoderma applanatum were used, the recoveries of the cellulose increased up to 29%. The same experiments were done by using a non-bred control strain, showing the recoveries of the cellulose from the treated or cultured rice straw to be 8%. PMID- 11866109 TI - Effects of heat and high-pressure treatments on antigenicity of beef extract. AB - The sera of bovine gamma globulin (BGG) positive beef allergic patients were used in this study in order to investigate changes in IgE-specific binding activity with regard to beef extract altered by heat or high-pressure treatment. In inhibition-ELISA, the sample treated at 60 degrees C did not show any significant changes in the antigenicity of BGG, but the sample treated at 100 degrees C showed a decrease of the antigenicity. In the case of the treatment with heating at 100 degrees C, heat-coagulation occurred in the beef extract. The resulting supernatant and precipitate of the sample by centrifugation were analyzed by immunoblotting. Only the fraction of precipitate showed a specific binding activity with the sera. Based on this result, it was speculated that the persistent antigenicity found even after the treatment at 100 degrees C in inhibition-ELISA remained principally in the heat-coagulated fraction, which indicated the importance of the method of handling the heat-coagulation in heat treatment. High-pressure treatments (200 MPa-600 MPa) of beef extract did not show any significant changes in the binding with the sera. PMID- 11866110 TI - A major decomposition product, citrinin H2, from citrinin on heating with moisture. AB - Citrinin is one of the mycotoxins produced by Penicillium citrinum. We examined the decomposition products after heating citrinin in water at 140 degrees C and isolated a major product, citrinin H2 (3-(3,5-dihydroxy-2-methylphenyl)-2 formyloxy-butane). Citrinin H2 did not show significant cytotoxicity to HeLa cells up to a concentration of 200 microg/ml (% cytotoxicity: 39%) in 63 h of incubation, but citrinin showed severe toxicity at a concentration of 25 microg/ml (% cytotoxicity: 73%). HPLC analysis of citrinin after heating under various conditions indicates that citrinin H2 is mainly yielded from citrinin. PMID- 11866111 TI - TMSCl as a mild and effective source of acidic catalysis in Fischer glycosidation and use of propargyl glycoside for anomeric protection. AB - Practical Fischer glycosidation was effected at room temperature or 60 degrees C by using 5 to 10 equiv. of TMSCl. The anomeric propargyl group formed by this method was found to be a versatile new protecting group, being stable in neat TFA but readily cleaved by treatment with Co2(CO)8 and TFA in CH2Cl2 via the formation of an alkyne-Co complex. PMID- 11866113 TI - Decrease in cytoplasmic pH-homeostastatic activity of the alkaliphile Bacillus lentus C-125 by a cell wall defect. AB - Cytoplasmic pH homeostatic activities of cell wall-defective derivatives of the alkaliphile Bacillus lentus C-125 were assessed using a pH-sensitive fluorescent probe, BCECF. It was shown that the acidic cell wall components took part in maintenance of the cytoplasmic pH neutrality at alkaline pH. PMID- 11866112 TI - Increased antibody production by retinoids is related to the fusion partner of human hybridomas. AB - An increase of human monoclonal antibody production caused by retinyl acetate and retinoic acid was influenced by the fusion partner rather than the original B lymphocyte used for the human hybridoma generation. Retinoid response of human hybridomas may be at least related to retinoid X receptor-alpha gene expression, which seemed to originate from their fusion partner. PMID- 11866114 TI - Inhibitory effect of isoflavones on peroxynitrite-mediated low-density lipoprotein oxidation. AB - Peroxynitrite, a potent oxidant formed in vivo from the reaction of nitric oxide with superoxide, can mediate low-density liprotein (LDL) oxidation which is thought to increase the risk of atherosclerosis. This study investigates the inhibitory effect of the isoflavones, genistein and daidzein, together with their glycosidic forms, genistin and daidzin, on the peroxynitrite-mediated LDL oxidation and nitration of tyrosine. Genistein and daidzein were observed to dose dependently inhibit peroxynitrite-mediated LDL oxidation, while their glucoside conjugates showed less activity. Moreover, all the isoflavones used in this study were found to be potent peroxynitrite scavengers, preventing the nitration of tyrosine. The ability of the isoflavones at 50 microM to decrease the tyrosine nitration induced by peroxynitrite (1 mM) was in the ratios of genistein (49%), daidzein (40%), daidzin (41%) and genistin (42%) when compared to the control (tyrosine incubated only with peroxynitrite). These results suggest that an intake of isoflavones could contribute to protecting against cardiovascular diseases and chronic inflammatory diseases. PMID- 11866116 TI - New antioxidative phenolic glycosides isolated from Kokuto non-centrifuged cane sugar. AB - Nine compounds, 3-hydroxy-4,5-dimethoxyphenyl-beta-D-glucopyranoside (1), beta-D fructfuranosyl-alpha-D-(6-vanilloyl)-glucopyranoside (2), beta-D-fructfuranosyl alpha-D-(6-syringyl)-glucopyranoside (3), 3-hydroxy-1-(4-hydroxy-3-methoxyphenyl) 2-[4-(3-hydroxy-1-(E)-propenyl)-2-methoxyphenoxy]propyl-beta-D-glucopyranoside (4), 3-hydroxy-1-(4-hydroxy-3-methoxyphenyl)-2-[4-(3-hydroxy-1-(E)-propenyl)-2,6 dimethoxyphenoxy] propyl-beta-D-glucopyranoside (5), dehydrodiconiferyl alcohol 9'-beta-D-glucopyranoside (6), 4-[ethane-2-[3-(4-hydroxy-3-methoxyphenyl)-2 propen]oxy]-2,6-dimethoxyphenyl-beta-D-glucopyranoside (7), 4-[ethane-2-[3-(4 hydroxy-3-methoxyphenyl)-2-propen]oxy]-2-methoxyphenyl-beta-D-glucopyranoside (8), and 3-hydroxy-1-(4-hydroxy-3,5-dimethoxyphenyl)-2-[4-(3-hydroxy-1-(E) propenyl)-2,6-dimethoxyphenoxy]propyl-beta-D-glucopyranoside (9), were isolated from Kokuto non-centrifuged cane sugar. Their structures were elucidated by spectroscopic evidence, mainly based on the NMR technique. Among them, seven new glycosides were identified. The 2-deoxyribose oxidation method was used to measure their antioxidative activity. All of these compounds showed antioxidative activities. PMID- 11866115 TI - Preparation and catalytic performance of lipases encapsulated in sol-gel materials. AB - Three kinds of lipases (from Candida antarctica, Pseudomonas cepacia, and Pseudomonas fluorescens) were encapsulated in inorganic matrices by the sol-gel method in order to synthesize chiral compounds by kinetic resolution. Sol-gel lipases prepared with vinyltriethoxysilane had higher hydrolysis activity for 2 octyl acetate than those with other silane precursors: tetramethoxysilane, methyltrimethoxysilane, and propyltrimethoxysilane. PMID- 11866117 TI - Effect of carnosine and related compounds on the inactivation of human Cu,Zn superoxide dismutase by modification of fructose and glycolaldehyde. AB - Glycolaldehyde, an intermediate of the Maillard reaction, and fructose, which is mainly derived from the polyol pathway, rapidly inactivate human Cu,Zn-superoxide dismutase (SOD) at the physiological concentration. We employed this inactivation with these carbonyl compounds as a model glycation reaction to investigate whether carnosine and its related compounds could protect the enzyme from inactivation. Of eight derivatives examined, histidine, Gly-His, carnosine and Ala-His inhibited the inactivation of the enzyme by fructose (p<0.001), and Gly His, Ala-His, anserine, carnosine, and homocarnosine exhibited a marked protective effect against the inactivation by glycolaldehyde (p<0.001). The carnosine-related compounds that showed this highly protective effect against the inactivation by glycolaldehyde had high reactivity with glycolaldehyde and high scavenging activity toward the hydroxyl radical as common properties. On the other hand, the carnosine-related compounds that had a protective effect against the inactivation by fructose showed significant hydroxyl radical-scavenging ability. These results indicate that carnosine and such related compounds as Gly His and Ala-His are effective anti-glycating agents for human Cu,Zn-SOD and that the effectiveness is based not only on high reactivity with carbonyl compounds but also on hydroxyl radical scavenging activity. PMID- 11866118 TI - Cloning and overexpression of Bacillus cereus penicillin-binding protein 3 gene in Escherichia coli. AB - The pbp3 gene encoding PBP3 of Bacillus cereus was cloned and sequenced. For this purpose, PBP3 was first purified from B. cereus ts-4, and N-terminal amino acid sequences of the peptides obtained from the protease digests of the protein were analyzed. The B. cereus ts-4 pbp3 gene consisted of an open reading frame of 1,986 bp encoding 662 amino acid residues with a calculated molecular mass of 73,044 Da. The active site-motifs SXXK, SXN, and KTG are present at the positions 393, 452, and 590, respectively, in the deduced amino acid sequence. The pbp3 structural gene was ligated into the pET17 x b expression vector and pET-pbp3 was constructed. A protein was produced by the cells of E. coli carrying pET-pbp3. The produced protein migrated at about 75 kDa in SDS-polyacrylamide gel and strongly reacted with biotinylated ampicillin. PMID- 11866119 TI - Leaf-closing substance in Leucaena leucocephala. AB - Potassium (2R,3R)-2,3,4-trihydroxy-2-methylbutanoate (1) was identified as a leaf closing substance in the nyctinastic plant, Leucaena leucocephala. Compound 1 showed strong leaf-closing activity toward L. leucocephala and was not effective against other nyctinastic plants. The potassium ion was indispensable for the bioactivity of 1. Compound 1 gradually lost its bioactivity because of the exchange of the counter cation during isolation. A leaf-opening substance was also observed in the same plant. PMID- 11866120 TI - Purification and properties of membrane-bound D-sorbitol dehydrogenase from Gluconobacter suboxydans IFO 3255. AB - D-Sorbitol dehydrogenase was solubilized from the membrane fraction of Gluconobacter suboxydans IFO 3255 with Triton X-100 in the presence of D sorbitol. Purification of the enzyme was done by fractionation with column chromatographies of DEAE-Cellulose, DEAE-Sepharose, hydroxylapatite, and Sephacryl HR300 in the presence of Triton X-100. The molecular mass of the enzyme was 800 kDa, consisting of homologous subunits of 80 kDa. The optimum pH of the enzyme activity was 6.0, and the optimum temperature was 30 degrees C. Western blot analysis suggested the occurrence of the enzyme in all the Gluconobacter strains tested. PMID- 11866121 TI - Polyphenol increases in safflower and cucumber seedlings exposed to strong visible light with limited water. AB - To assess effects of the environmental stress on polyphenol compounds (polyphenols) in plants, the polyphenol contents were investigated in the seedlings of safflower (Carthamus tinctrius L.) and cucumber (Cucumis sativus L.) grown under three types of growth conditions: control; light stress, irradiated with strong light in the visible wavelength range; and light/water stress, irradiated with strong visible light with a limited water supply. The total polyphenol contents and the amounts of the major polyphenols, especially luteolin 7-O-glucoside in safflower cotyledons, and luteolin 7-O-glucoside and luteolin in safflower foliage leaves, increased in response to both stresses. The polyphenol increasing effect of light/water stress was clearly observed in safflower compared to cucumber, suggesting that plants that are resistant to these stresses can accumulate substantial amounts of polyphenols compared to the plants which respond weakly to the stresses. PMID- 11866122 TI - Kinetic expression for the oxidation of linoleic and arachidonic acid esters in their mixed system. AB - Two polyunsaturated fatty acids (PUFAs) or their esters were mixed, and their oxidation processes were measured at 65 degrees C and ca. 0% relative humidity. Except when a PUFA ester was mixed with a free PUFA, the oxidation of the less oxidative PUFA was promoted as its content in the mixture decreased, while the oxidation of the more-oxidative PUFA was delayed with a decrease in its content. A kinetic model is proposed whereby a PUFA acts as the diluent for another PUFA, and the oxidation rate of the PUFA is proportional to the product of the unoxidized PUFA concentration and the sum of the concentrations of the oxidized PUFA and the other oxidized PUFA. This model well expressed the oxidation processes of the PUFAs in their mixed system. PMID- 11866123 TI - Diverse bacteria related to the bacteroides subgroup of the CFB phylum within the gut symbiotic communities of various termites. AB - Phylogenetically diverse clones of the partial 16S rDNA (ca. 850 bp) of bacteria belonging to the bacteroides subgroup of the cytophaga-flavobacter-bacteroides phylum were collected from the symbiotic microbial communities in the guts of six termite species without cultivation. Combined with the sequences reported previously, a total of thirty phylotypes of the subgroup were identified and classified into five phylogenetic clusters. One that was comprised of the phylotypes from a single termite species was related to the genus Rikenella. Two were clustered each with some cultured strains, genera of which have not been clearly defined yet. The remaining two clusters had no culturable representatives, suggesting the presence of yet-uncultivated genera within the termite guts. From these sequence data, we designed a specific primer for the bacteroides subgroup, which was successful in the terminal-restriction fragment length polymorphism analysis to detect the phylotypes of the subgroup in the termite gut. PMID- 11866124 TI - Effects of GroESL coexpression on the folding of nicotinoprotein formaldehyde dismutase from Pseudomonas putida F61. AB - The overexpression of fdm, which encodes the formaldehyde dismutase from Pseudomonas putida F61, resulted in the formation of inclusion bodies made up of aggregated enzyme, leaving little activity in the soluble fraction of the transformant cells. On the other hand, coexpression of groESL along with fdm facilitated in vivo solubilization of the enzyme protein in its active form. When coexpressed with groESL, formaldehyde dismutase purified from E. coli had the same crystalline form (i.e., a regular octahedron) as the native enzyme, and like the native enzyme, it bound 1 mol of NAD(H) and 2 mol of zinc in each subunit. PMID- 11866125 TI - Effects of dietary protein of proso millet on liver injury induced by D galactosamine in rats. AB - In this paper, we examined the effects of dietary protein from proso millet on liver injury induced by D-galactosamine or carbon tetrachloride in rats using serum enzyme activities as indices. D-galactosamine-induced elevations of serum activities of aspartate aminotransferase, alanine aminotransferase, and lactate dehydrogenase were significantly suppressed by feeding the diet containing 20% protein of proso millet for 14 days as compared with those of rats fed a 20% casein diet, but not in the case of carbon tetrachloride. The results showed that proso millet protein is effective at lower dietary protein levels than that of dietary gluten reported previously. Therefore, the findings reported here may suggest that proso millet protein is considered to be another preventive food for liver injury. PMID- 11866126 TI - A novel protein toxin from the deadly box jellyfish (Sea Wasp, Habu-kurage) Chiropsalmus quadrigatus. AB - The deadly box jellyfish (Sea Wasp, Habu-kurage in Japanese) Chiropsalmus quadrigatus Haeckel (Cubozoa) is distributed widely in the tropical Pacific region. In Japan, three fatal cases due to stings from this species have been reported officially. We successfully isolated C. quadrigatus toxin-A (CqTX-A, 44 kDa), a major proteinaceous toxin, for the first time, from the nematocysts of C. quadrigatus. CqTX-A showed lethal toxicity to crayfish when administered via intraperitoneal injection (LD50 = 80 microg/kg) and hemolytic activity toward 0.8% sheep red blood cells (ED50 = 160 ng/ml). Furthermore, we sequenced the cDNA encoding CqTX-A. The deduced amino acid sequence of CqTX-A (462 amino acids) showed 25.2% and 21.6% sequence similarity to Carybdea rastoni toxins (CrTXs) and Carybdea alata toxin-A (CrTX-A), respectively, which are Cubozoan jellyfish toxins. PMID- 11866128 TI - The effects of 3-methylcholanthrene on macrophage respiratory burst and biotransformation activities in the common carp (Cyprinus carpio L.). AB - The sensitivity of phagocytic cell function as a bioindicator of pollution stress by polycyclic aromatic hydrocarbons was evaluated in the common carp (Cyprinus carpio L). The time course response of the head-kidney macrophage respiratory burst was measured 1, 2, 3, 5 and 7 days after intraperitoneal injection of a prototypical Cyp 1A inducer (3-methylcholanthrene). This immune activity was compared to the rate of induction of total cytochrome P450, ethoxyresorufin O deethylase activity (EROD) and glutathione S-transferase activity (GST) in the liver and head-kidney. 3-methylcholanthrene (40 mg kg(-1)) caused a rapid increase in the macrophage respiratory burst. This response was maximal at day 3 post exposure and coincided with maximum induction of cytochrome P450 and EROD activity in liver and head-kidney. Moreover, alpha-naphtoflavone, which functions as both an Ah receptor antagonist and an inhibitor of cytochrome P450 1A activity, reversed the 3-methylcholanthrene induction of immune and enzymatic parameters measured, suggesting metabolic processes. Taken together these results suggest that the induction of macrophage oxidative function may be an equally sensitive marker of exposure to polycyclic aromatic hydrocarbon as the induction of biotransformation activities and confirm that responses mediated by the Ah receptor are similar, if not identical, to those of mammals. PMID- 11866127 TI - The effect of dietary immunomodulation upon Edwardsiella tarda vaccination in healthy and immunocompromised Indian major carp (Labeo rohita). AB - In order to determine the impact on disease resistance of four dietary immunomodulators viz., beta-1,3 glucan, levamisole, vitamins C and E, in an important farmed Indian major carp species, rohu (Labeo rohita Ham.), fish were fed diets containing various levels of these substances during a 60 day trial. Aflatoxin B1 (AFB,) at 125 mg kg(-1) body weight was injected intraperitoneally (i.p.) into fish to induce an immunosuppressive state on the first day of the experiment in some individuals. The fish were vaccinated against formalin-killed Edwardsiella tarda vaccine on day 30 of the experiment. Specific immunity, as measured by bacterial agglutination titre and disease resistance against E. tarda, was determined at the end of the trial. The results demonstrate that all the four immunomodulators were capable of significantly (P<0.05) increasing specific immunity and reducing mortality in immunocompromised fish but failed to enhance specific immunity and protection in healthy fish. The increased bacterial agglutination titre by beta-1,3 glucan, and reduced mortality losses by both beta 1,3 glucan and levamisole were marked in healthy vaccinated fish compared with their controls. Similarly, all four substances significantly reduced the mortality rates in immunocompromised and healthy unvaccinated fish. Out of these four substances, glucan was recorded to be the most effective immunomodulator in rohu. The present results suggest that the introduction of these substances into the diet of fish grown in farms under immunosuppressive/stressful conditions could increase their resistance to infection by reducing mortality rates and offer economic benefits. PMID- 11866129 TI - Temperature dependent activation of leucocyte populations of rainbow trout, Oncorhynchus mykiss, after intraperitoneal immunisation with Aeromonas salmonicida. AB - The temperature dependence of in vivo activation of rainbow trout Oncorhynchus mykiss, leucocyte populations after intraperitoneal injection (i.p.) of fish with a T-cell independent antigen Aeromonas salmonicida (strain MT423) was investigated using a proliferation assay and flow cytometric analysis with mab specific for trout leucocyte surface markers. In trout kept at 15-17 degrees C a prominent activation of blood and spleen leucocytes was found. Also, drastic changes of the percentage of the leucocyte populations in blood and spleen occurred: the amount of monocytes in the blood increased between day 2 and day 7 post injection (p.i.), whereas in spleen the amount of monocytes stayed at a high level (approximately 35%) after a depression between day 4 and day 7 p.i. The percentage of B-lymphocytes was increased first in spleen and then in blood. The percentage of granulocytes in blood was elevated during the whole experiment compared to control fish. In trout kept at 10-12 degrees C only blood leucocytes showed a weak activation after i.p. injection of A. salmonicida, whereas spleen leucocytes showed nearly no reaction. Only the percentage of granulocytes in the blood (day 2-14 p.i.) and of monocytes in the spleen (day 2 and day 8 p.i.) was changed compared to phosphate buffered saline (PBS)-injected fish. However, the development of A. salmonicida specific antibodies was contrary to the cellular reaction. Whereas antibodies could first be detected after 16-18 days p.i. in both groups the amount of antibodies was significantly higher in sera of trout kept at 10-12 degrees C at day 22 and day 28 p.i. than in sera of trout kept at 15-17 degrees C. These results indicate stronger A. salmonicida induced activation of monocytes, granulocytes and B-lymphocytes at higher temperature. However, the development of a specific antibody response against A. salmonicida seemed to be more effective at lower temperatures. PMID- 11866130 TI - Effects of four anaesthetics on the innate immune response of gilthead seabream (Sparus aurata L.). AB - Anaesthesia may depress the immune system in mammals, but there is no available information on this topic in fish. In the present work, four anaesthetics that are used in aquaculture, MS222 (0 19 mM), benzocaine (0.21 mM), 2-phenoxyethanol (16 mM) and quinaldine sulphate (0.083 mM), were tested in order to observe their effects on the gilthead seabream (Sparus aurata L.) innate immune system. The results showed that the four anaesthetics produced increased blood glucose levels after an hour. In addition, benzocaine and 2-phenoxyethanol depressed complement activity and phagocytosis, while MS222 and quinaldine sulphate did not. Some anaesthesia is a common practice in aquaculture, the data obtained should be taken into account to avoid possible immunodepression in farmed fish. PMID- 11866131 TI - The effect of temperature on non-specific defence parameters of three strains of juvenile Atlantic halibut (Hippoglossus hippoglossus L.). AB - The effect of temperature (8, 12, 15 and 18 degrees C) on a variety of non specific defence and haematological parameters was examined in three geographically distinct reared strains (Canadian, Icelandic, Norwegian) of Atlantic halibut. The results indicate that temperature exerts a considerable influence on some blood parameters (packed cell volume and the percentage population of leucocytes in peripheral blood) and on some humoral parameters (serum lysozyme activity and serum protein levels) of halibut. A high temperature of 18 degrees C caused a decrease in the number of circulating blood cells and an increase in serum lysozyme levels; effects consistent with those reported within the literature for stress. The different strains of halibut exhibited differing responses with respect to differential counts of peripheral blood lymphocytes and thrombocytes, and to serum protein concentrations, serum lysozyme activity, serum iron content, unsaturated iron binding capacity of serum and O2- production by kidney macrophages. PMID- 11866132 TI - Pathogenesis and immune response in Atlantic salmon (Salmo salar L.) parr experimentally infected with salmon pancreas disease virus (SPDV). AB - Atlantic salmon parr were injected intraperitoneally with salmon pancreas disease virus (SPDV) grown on CHSE-214 cells. The viraemia, the histopathological changes in target organs and some immune parameters were taken at intervals up to 30 days post-infection (dpi). The earliest kind of lesion was necrosis of exocrine pancreas, appearing as soon as 2 dpi. It progressed towards complete tissue breakdown at 9 dpi before resolving gradually. Concurrent to this necrosis, a strong inflammatory response was in evidence from 9 dpi in the pancreatic area for a majority of fish. A necrosis of the myocardial cells of the ventricle occurred in infected fish mainly at 16 dpi and it faded thereafter. The monitoring of the plasma viral load showed a rapid haematogenous spreading of SPDV, peaking at 4 dpi, but also the absence of a secondary viraemia. No interferon (IFN) was detected following the infection of parr with SPDV, probably owing to an IFN activity in Atlantic salmon below the detection level of the technique. Neutralising antibodies against SPDV were in evidence from 16 dpi and they showed a time-related increasing titre and prevalence. The phagocytic activity in head-kidney leucocytes was always significantly higher in the infected fish than in the control fish, being particularly high by 9 dpi. Lysozyme and complement levels were both increased and they peaked significantly in the infected fish at 9 and 16 dpi respectively. These results demonstrated that an experimental infection of Atlantic salmon parr with SPDV provoked a stimulation of both specific and non-specific immunity with regards to the viraemia and the histopathology. PMID- 11866134 TI - Current status of genetic testing for colorectal cancer susceptibility. AB - Over 130,000 new cases of colon cancer are diagnosed annually. Approximately 20% to 30% of these are attributable to familial risk, and 3% to 5% belong to a hereditary colorectal cancer predisposition syndrome. Recent discoveries of the genes responsiblefor the inherited colorectal cancer conditions have expanded the field of commercial genetic testing. Health-care providers who use genetic testing in clinical practice are aware of the benefits that genetic testing can confer on screening, prevention, and treatment options for patients with a personal and/or family history of colon cancer. When genetic test results are correctly interpreted, the information they provide can offer medical guidance for the entire family. The psychological impact, however, of presymptomatic testing can be multifaceted. There are unprecedented benefits but also complex issues surrounding genetic testing. For these reasons, the practice of offering genetic testing to individuals at high risk for colon cancer is heavily fortified with guidelines and recommendations. This review covers the current availability and limitations of genetic testing for inherited colorectal cancer syndromes and focuses on guidelines that address the psychological, ethical, and social concerns stemming from genetic testing. PMID- 11866133 TI - State tobacco settlement funds not being spent on vigorous tobacco control efforts. PMID- 11866135 TI - Surgery in the older patient. AB - Surgery is still the most important treatment for solid tumors, regardless of the age of the patient. In this article, we discuss the physiology of aging as it relates to risk assessment in the elderly surgical oncology patient. A brief review of the role of surgery in the treatment of breast, colorectal, pancreatic, and gastric cancer is provided, because these solid tumors primarily affect elderly patients. Options for palliation are discussed. We conclude that older patients should not be deprived of curative surgery based on chronologic age alone. PMID- 11866136 TI - Clinical trials referral resource. Clinical trials and NCI resources for cancer in HIV-positive patients. PMID- 11866137 TI - Chronic inflammation and cancer. AB - A substantial body of evidence supports the conclusion that chronic inflammation can predispose an individual to cancer, as demonstrated by the association between chronic inflammatory bowel diseases and the increased risk of colon carcinoma. Chronic inflammation is caused by a variety of factors, including bacterial, viral, and parasitic infections, chemical irritants, and nondigestible particles. The longer the inflammation persists, the higher the risk of associated carcinogenesis. This review describes some of the underlying causes of the association between chronic inflammation and cancer. Inflammatory mediators contribute to neoplasia by inducing proneoplastic mutations, adaptive responses, resistance to apoptosis, and environmental changes such as stimulation of angiogenesis. All these changes confer a survival advantage to a susceptible cell. In this article, we discuss the contribution of reactive oxygen and nitrogen intermediates, prostaglandins, and inflammatory cytokines to carcinogenesis. A thorough understanding of the molecular basis of inflammation associated neoplasia and progression can lead to novel approaches to the prevention and treatment of cancer. PMID- 11866138 TI - Carcinomatous Meningitis: it does not have to be a death sentence. AB - Carcinomatous meningitis, specifically leptomeningeal metastases from solid tumors, has a dismal prognosis, with an overall median survival of 2 to 4 months. Lymphomatous meningitis has a better outlook, with a median survival of more than 6 months, but diagnosis may be delayed and treatment is not curative. Despite these poor statistics, there are subsets of patients who do relatively well. Moreover, there are a number of new approaches to the diagnosis and treatment of leptomeningeal metastases that promise to extend life and prevent disability. These include molecular techniques of diagnosis, expanding the repertoire of drugs available for intrathecal administration, using systemic chemotherapy to treat leptomeningeal disease, and applying strategies such as gene therapy and immunotoxins to the management of leptomeningeal metastases. These novel approaches offer the hope of liberating patients from a death sentence and providing clinicians with effective weapons in the fight against a dreaded neurologic complication of systemic cancer. PMID- 11866139 TI - Metastatic bone disease. PMID- 11866140 TI - Health care is a privilege, not a right. PMID- 11866141 TI - Health care is a privilege, not a right. PMID- 11866142 TI - Shoulder arthroplasty in the patient with a deficient rotator cuff: an old concept reinvented. PMID- 11866143 TI - Radiologic case study. Pyogenic left sacroiliac infection. AB - A wide variety of inflammatory, infectious, metabolic, and neoplastic diseases can affect the sacroiliac joints. Early in their course, some of these diseases produce only subtle and nonspecific signs and symptoms. Demonstration of characteristic joint changes, knowledge of specific clinical manifestations, and familiarity with imaging modalities can lead to early diagnosis and treatment of these patients. PMID- 11866144 TI - Volar neutralization plate fixation of dorsally displaced distal radius fractures. AB - Open reduction and volar neutralization plate fixation avoids problems associated with dorsal plate fixation (eg, extensor tenosynovitis and tendon rupture) and minimizes the necessity for secondary plate removal. Volar plating is a useful technique for treating dorsally displaced distal radius fractures with minimal scarring, infrequent bone grafting, and safe early wrist mobilization. PMID- 11866145 TI - Early results of a reverse design prosthesis in the treatment of arthritis of the shoulder in elderly patients with a large rotator cuff tear. AB - Results of shoulder arthroplasty in patients with a deficient rotator cuff often are suboptimal with significant limitations in postoperative active mobility. Short-term results using a reverse design prosthesis in the treatment of the cuff deficient arthritic shoulder are encouraging. This prosthesis compares favorably, particularly with regard to postoperative active anterior elevation, to other treatment options in this challenging patient population. PMID- 11866146 TI - Use of an allograft bone button for rotator cuff repair. AB - Bone button anchors constructed from allograft bone were investigated as an alternative for rotator cuff repair. Bone button anchors were used in 59 consecutive procedures after mechanical pullout strength and cyclical loading bench testing. Sequential radiographs demonstrated that the anchors locked in cancellous bone and provided consistent fixation with no loosening or lysis in 1 year of clinical follow-up. Mechanical testing demonstrated mean, pullout strength of 160.2 N (36 lbs), and the anchors did not migrate when cyclically loaded. Clinical results showed an allograft anchor is viable for rotator cuff tendon repair. PMID- 11866147 TI - One-stage bilateral total hip arthroplasty in patients > or = 75 years. AB - Since 1991, 43 one-stage sequential bilateral total hip arthroplasties (THAs) inpatients > or = 75 years have been performed by the authors. The results were assessed to determine whether thiss procedure can be performed safely and with good outcome in this subset of patients. These results were compared to those of 1 36 patients < 75 years who underwent the same procedure. The one-stage bilateral procedure was performed in all patients who had signiificant arthritic disease of both hips. There was no diffference in length of surgery or length of hospital stay. Patients > or = 75 years were more likely to experience minor postoperative morbidity such as ileus and urinary tract infections. The incidence of myocardial ischemia was 2.3% in the elderly group With a mean follow-up of 2.5 years, 90% of the elderly patients lived independently, 80% walked without assistive devices, and 90% were pain free. Therefore, one-stage sequential bilateral THA is a safe and effective option for patients > or = 75 years when pain and functional limitations affect quality of life. PMID- 11866148 TI - A level one trauma center's experience with the posterior approach to the pelvis. AB - A retrospective review of the experience at Los Angeles County and University of Southern California Medical Center was conducted as part of a multicenter study to evaluate the true complication rate of the posterior approach to the sacroiliac joint. Between 1995 and 1997, 35 patients underwent 42 approaches, representing all patients who underwent the posterior approach by a single staff surgeon at these medical centers. All patients underwent follow-up > or = 1 year postoperatively. (11%) neurologic complications were found postoperatively, all of which resolved prior to discharge. There was 1 (2.4%) wound complication. There was 1 gluteal flap for closure (open fracture) and 1 secondary wound closure. Five patients had prolonged wound drainage (> 5 days). There were no skin sloughs in the series and only 1 patient developed a deep wound infection. Contrary to reports by advocates of anterior approaches and closed reductions, the posterior approach allows anatomic reduction of posterior lesions with an acceptable complication rate. PMID- 11866149 TI - Vertebral body fracture of the lumbar spine in elderly women: more severe in osteoarthritis of the knee than in femoral neck fracture. AB - This study investigated bone fragility by comparing fractures of the vertebral body of the spine in elderly women receiving total knee replacement (TKR) (group 1) due to severe osteoarthritis of the knee and those with femoral neck fractures (group 2) attributable to osteoporosis. Forty-two women each were selected retrospectively for group 1 and prospectively for group 2. Patient age ranged from 64-83 years. Vertebral body fractures of the lumbar spine were significantly more severe in group 1 than in group 2 (P<.001). Patients undergoing TKR due to osteoarthritis of the knee had systemic bone fragility, which included the spine. PMID- 11866150 TI - Distal radius fractures in adults. AB - Although distal radius fractures are a common injury in the elderly and young adult population, the classification, treatment options, and assessment of outcomes of these fractures remain controversial. Since there is no uniform fracture classification system, it is difficult to compare studies. An evidence based model of management needs to be developed. PMID- 11866151 TI - Identical pediatric chance fractures in identical twins. PMID- 11866152 TI - Multiple gamma nail failure. PMID- 11866153 TI - Floating elbow in an infant. PMID- 11866155 TI - The clinical history and development of the low contact stress total knee arthroplasty. AB - Contemporary total knee arthroplasty (TKA) has evolved over the past 30 years as a result of early prosthetic design failure. Hinge-type implants demonstrated a high loosening rate because of overconstraint. Curved-on-flat prostheses, which were popular in the late 1970s, became associated with a high incidence of polyethylene failure. The Low Contact Stress (LCS) Total Knee System (DePuy Orthopaedics Inc, Warsaw, Ind), a mobile-bearing knee arthroplasty design, was introduced in 1977. The LCS Total Knee System, now in its 25th year without alterations in articular geometry, has demonstrated clinical success and has had an impact on evolving prosthetic designs. Introduced in 2001, the LCS Complete Total Knee System provides further refinements in patient fit without changing the fundamental design principles. Surgeons are cautioned to use a well-proven clinical design, as biomechanical variations critically impact clinical outcomes. PMID- 11866154 TI - Popliteal pseudoaneurysm after total knee arthroplasty: MRI of the vascular anatomy. PMID- 11866156 TI - World experience with low contact stress mobile-bearing total knee arthroplasty: a literature review. AB - This article reviews recent data regarding the low contact stress (LCS) mobile bearing total knee prosthesis recently published in the literature; conclusions are based on literature controls. The LCS Total Knee System (DePuy Orthopaedics Inc, Warsaw, Ind) has addressed the issues of implant polyethylene wear and implant-generated osteolysis. Patellar implant complications have been minimal despite the use of a mobile, metal-backed component. The original US Food and Drug Administration clinical investigation demonstrated favorable experience with cementless fixation. Results from numerous series indicate that the main problem with a mobile-bearing device is an early prosthetic failure resulting from bearing dislocation or breakage. Inadequate surgical technique will lead to femorotibial malalignment or ligamentous instability, which are factors that commonly result in revision. With the LCS Total Knee System, surgeons can use a tibia-cut-first approach with careful ligamentous balancing and flexion block spacing. The valgus lateral knee approach is an additional technique that has proved useful with this method. PMID- 11866157 TI - In vivo kinematics of the low contact stress rotating platform total knee. AB - The Rocky Mountain Musculoskeletal Research Laboratory uses in vivo video fluoroscopy to evaluate kinematics of total knee arthroplasty. This method allows us to evaluate in vivo weight bearing activity for a variety of knee designs, as well as kinematics of the normal knee. Our proprietary method of automated model fitting allows for three-dimensional analysis obtained from two-dimensional fluoroscopic images with minimal error. This article outlines the details of our kinematic analysis of the Low Contact Stress (LCS) Total Knee System (DePuy Orthopaedics lnc, Warsaw, Ind) rotating platform total knee and compares this design with fixed-bearing posterior cruciate-retaining and posterior cruciate substituting total knee and normal knee kinematics. PMID- 11866158 TI - Polyethylene wear in mobile-bearing prostheses. PMID- 11866159 TI - Factors affecting polyethylene wear in total knee arthroplasty. AB - A complication of total knee arthroplasty (TKA) is fatigue-type wear, which can destroy a tibial inlay in <10 years. This deleterious wear mechanism occurs during cyclic loading if the yield stress of polyethylene is exceeded. Because increased stress on and within the polyethylene inlay is associated with increased wear, it is important to reduce the inlay stress by either activity restrictions or conformity changes of design. All stress parameters are more sensitive to conformity changes (eg, design changes) than to load changes (eg, activity restrictions). However, the reduction of stress on and within the polyethylene through increased conformity will increase the stress at the tibial fixation interfaces. An attempt was made to solve this problem with the introduction of mobile-bearing designs. Many mobile-bearing designs exist with good long-term results. One important difference among the various designs is the amount of flexion range with full conformity between the femoral component and the tibial inlay. Although a single radius design reduces polyethylene stress throughout the flexion range, it may be disadvantageous for a revision design to intraoperatively adapt to different degrees of constraint. Aseptic loosening and osteolysis due to small abrasive and adhesive wear particles have also been reported as a cause of failure. The design and material parameters affecting polyethylene wear in TKAs, as well as the potential detrimental effects of wear particle size, are the key issues in defining the life of a TKA. PMID- 11866160 TI - The influence of sterilization method on wear performance of the low contact stress total knee system. AB - Sterilization method changes to the Low Contact Stress (LCS) Total Knee System (DePuy Orthopaedics Inc, Warsaw, Ind) have resulted in progressive improvements in wear resistance. Initially, the system used the traditional gamma in air irradiation sterilization method, but was transitioned to Gas plasma sterilization. The Gas plasma technique did not introduce free radicals that subsequently oxidize with extended shelf aging found with gamma in air irradiation. This oxidation has demonstrated a significant reduction in mechanical properties leading to increased wear. The introduction of Gas plasma was also accompanied by a transition to polyethylene resins that no longer contained calcium stearate, surmised to influence the rate of oxidation. More recently, gamma irradiation and storage of the polyethylene components in an essentially oxygen-free environment imparted by gamma irradiation in a vacuum foil pouch (GVF), has been shown to protect the implant from oxidation. Additionally, the GVF process imparts the beneficial effects of moderate crosslinking that has been demonstrated to improve wear resistance. PMID- 11866161 TI - The clinical history of mobile-bearing patella components in total knee arthroplasty. AB - Two hundred fifty-six primary cementless meniscal-bearing total knee arthroplasties were performed between May 1985 and January 1989. All knees were replaced with a low contact stress metal-backed anatomic mobile patella. The average patient follow-up was 11.5 years. No patellae were revised for failure of fixation and no dislocations or subluxations were reported. One patella was revised for polyethylene wear, and one well-functioning component was removed at the time of revision to facilitate range of motion and wound closure. The survival estimate at 12 years was 99.5%. Compared to the high incidence of failure of metal-backed fixed-bearing patellae components, the anatomic rotating patella provides durable long-term results with a low incidence of complications. PMID- 11866162 TI - Fixed- versus mobile-bearing total knee arthroplasty: technical issues and surgical tips. AB - Mobile-bearing knee arthroplasties have been used clinically for 25 years. The success of this technology depends on strict adherence to the principles of flexion-extension gap technique. The use of the fixed femoral landmarks for the rotational positioning of the femoral component (measured resection technique) is not acceptable if bearing dislocation is to be avoided. The principles of flexion extension gap balancing, as well as the surgical technique, are reviewed in this article. PMID- 11866163 TI - Alternate surgical approaches in mobile-bearing total knee arthroplasty. AB - Surgical approaches in nonconstrained mobile-bearing total knee arthroplasty (TKA) should provide the best access for specific deformity correction. Alternatives to the standard medial approach include the medial midvastus and subvastus approaches, and the direct lateral approach, with or without tibial tubercle osteotomy or proximal "rectus snip." The direct lateral approach is a recommended alternative for fixed valgus knees, especially if there is lateral patellar subluxation and significant rotational deformity. The subvastus and midvastus medial approaches preserve quadriceps integrity and patella stability, and are recommended for severely unstable knees in nonobese, non-fixed valgus, and medial unicompartmental knee replacements. PMID- 11866164 TI - Advanced mobile-bearing surgical technique. AB - Consistent success in total knee arthroplasty (TKA) is dependent on achieving basic surgical goals that include a neutral mechanical axis, a restored joint line, and equally balanced flexion and extension gaps. Although these surgical goals apply to both fixed-bearing and mobile-bearing TKAs, concerns about bearing stability in mobile-bearing implants require surgeons to be aware of flexion gap stability. The increased awareness required to achieve flexion gap stability with the mobile-bearing implant not only provides stability but also encourages surgeons to adhere to basic surgical principles. PMID- 11866165 TI - Selective attention and emotional vulnerability: assessing the causal basis of their association through the experimental manipulation of attentional bias. AB - Although it is well-established that vulnerability to negative emotion is associated with attentional bias toward aversive information, the causal basis of this association remains undetermined. Two studies addressed this issue by experimentally inducing differential attentional responses to emotional stimuli using a modified dot probe task, and then examining the impact of such attentional manipulation on subsequent emotional vulnerability. The results supported the hypothesis that the induction of attentional bias should serve to modify emotional vulnerability, as revealed by participants' emotional reactions to a final standardized stress task. These findings provide a sound empirical basis for the previously speculative proposal that attentional bias can causally mediate emotional vulnerability, and they suggest the possibility that cognitive experimental procedures designed to modify selective information processing may have potential therapeutic value. PMID- 11866166 TI - The DRD4 VNTR polymorphism influences reactivity to smoking cues. AB - Recent research has indicated that craving for tobacco can be reliably elicited by exposure to smoking cues, suggesting that cue-elicited craving for tobacco may be a useful phenotype for research on genetic factors related to nicotine dependence. Given the potential role of dopamine in cue-elicited craving, the authors examined whether the DRD4 VNTR polymorphism is associated with cue elicited craving for tobacco. Participants who were homozygous or heterozygous for the 7 repeat (or longer) allele were classified as DRD4 L, and all other participants were classified as DRD4 S. Participants were exposed to smoking cues before smoking either high-nicotine cigarettes or control cigarettes. Analyses suggested that participants in the L group demonstrated significantly greater craving, more arousal, less positive affect, and more attention to the smoking cues than did the participants in the S group. PMID- 11866167 TI - Test of the plausibility of adolescent substance use playing a causal role in developing adulthood antisocial behavior. AB - DSM-IV antisocial personality disorder diagnosis requires that conduct disorder be exhibited before age 15. However, recent studies have reported on men and women without conduct disorder before age 15 but qualified for the adulthood antisocial personality criterion (AAB). This general-population, retrospective study investigated the plausibility of causal relationships between adolescent drug and alcohol misuse (ADAM) and AAB among subgroups who reported childhood onset conduct problems (CP), adolescent-onset CP, or no more than one conduct problem. Data from the Epidemiological Catchment Area Study (N = 8,724) suggested that persons with childhood-onset CP are at much greater risk for AAB than persons with adolescent-onset CP. Nevertheless, large proportions of men and women with AAB had adolescent-onset CP or no CP. Regardless of CP history, being drunk by age 18 or having a drug use-related symptom before age 18 increased AAB risk, even after controlling for having a substance use-related disorder in adulthood. Mechanisms that potentially explain these associations are discussed. PMID- 11866168 TI - Individual differences in the emergence of depressive symptoms in children and adolescents: a longitudinal investigation of parent and child reports. AB - The authors address questions about the rate that depressive symptoms emerge, developmental and gender differences in this rate, and differences between parent and child estimates of this rate. In a 12-wave, cohort-sequential, longitudinal design, 1,570 children (Grades 4-11) and parents completed reports about children's depression. Cross-domain latent growth curve analysis revealed that (a) the rate of symptom growth varied with developmental level. (b) gender differences symptom growth preceded emergence of mean level gender differences, (c) the rate of symptom development varied with age, and (d) parent-child agreement about rate of symptom change was stronger than agreement about time specific symptoms. The authors suggest that predictability of depressive symptoms varies with age and the dimension under investigation. PMID- 11866169 TI - Acculturation and peritraumatic dissociation in young adult Latino survivors of community violence. AB - This study examined the relationship between acculturation and peritraumatic dissociation in a sample of 304 physically injured Latino survivors of community violence. Item response theory analyses were conducted to document the measurement equivalence of English- and Spanish-language versions of a scale measuring peritraumatic dissociation. After establishing equivalence, structural equation modeling was used to determine the impact of acculturation on peritraumatic dissociation after controlling for other relevant covariates, including assault characteristics, intoxication before the assault, and trauma exposure history. Acculturation emerged as a significant and negative predictor of dissociation, so that high levels of acculturation were associated with low levels of peritraumatic dissociation. These findings offer a counterinstance to the emerging consensus that retention of Latin American cultural traditions serves to promote mental health. PMID- 11866170 TI - Directed forgetting in acute stress disorder. AB - The rationale underpinning the diagnosis of acute stress disorder is that cognitive mechanisms result in avoidant processing of aversive experiences. This study investigated acutely traumatized participants with either acute stress disorder (ASD; n = 15) or no ASD (n = 14) and nontraumatized comparison participants (n = 16). Participants were administered intermixed presentations on a computer screen of positive, neutral, and trauma-related words that were followed by instructions to either remember or forget each word. On a subsequent recall test, ASD participants displayed poorer recall of to-be-forgotten trauma related words than did non-ASD participants. Severity of psychopathology was negatively correlated with to-be-remembered positive words. These findings are consistent with the proposal that people who develop ASD display an aptitude for superior forgetting of aversive material. PMID- 11866171 TI - Distress tolerance and duration of past smoking cessation attempts. AB - The present study tested the hypothesis that limited ability to tolerate physical and psychological distress is associated with early relapse from smoking cessation. Specifically, the authors exposed 16 current smokers who had failed to sustain any previous quit attempt for more than 24 hr (immediate relapsers) and 16 smokers with at least 1 sustained quit attempt of 3 months or longer (delayed relapsers) to psychological (mental arithmetic) and physical (carbon dioxide inhalation-breath holding) stressors. Relative to delayed relapsers, immediate relapsers were characterized by higher baseline levels of affective vulnerability, by greater levels of dysphoria and urge to smoke after 12 hr of nicotine deprivation, and by less task persistence on the stressors, suggesting that these may be risk factors for early lapse in the context of quitting smoking. PMID- 11866172 TI - Emotional valence and reference disturbance in schizophrenia. AB - The authors examined the impact of emotional valence on reference disturbance in patients with schizophrenia and identified factors that moderate individual differences in this relationship. Thirty-nine patients with schizophrenia were given an interview that elicited speech samples in response to 10 neutral and 6 negatively valenced emotional questions. Speech samples were analyzed using the Communication Disturbances Index (N. M. Docherty, M. DeRosa, & N. C. Andreasen, 1996). Participants were also assessed with the Positive and Negative Symptom Scale (S. Kay, 1991) and a single-trial Stroop task (Stroop, 1935). Group analyses indicated that negative questions elicited more unclear references than neutral questions. Both the severity of disorganization symptoms and selective attention deficits predicted increased reference errors in response to emotional questions. PMID- 11866173 TI - Depressive symptoms and lymphocyte proliferation in older adults. AB - In an 18-month prospective study, community-dwelling older adults, including both spousal caregivers of dementia patients and noncaregiving controls, were examined. Participants were selected on the basis of the presence or absence of chronic depressive symptoms that exceeded a cutoff score for clinically relevant depressive symptoms on a self-report symptom measure. Compared with nondepressed older adults, those with chronic, mild depressive symptoms had poorer T cell responses to 2 mitogens from baseline to follow-up. Additionally, among individuals with depressive symptoms, older age was associated with the poorest blastogenic response to the mitogens at follow-up. These findings extend the association between depression and immune function to community-dwelling older adults with chronic, mild depressive symptoms. PMID- 11866174 TI - Reasons for suicide attempts and nonsuicidal self-injury in women with borderline personality disorder. AB - Self-reported reasons for suicide attempts and nonsuicidal self-injury were examined using the Parasuicide History Interview within a sample of chronically suicidal women meeting criteria for borderline personality disorder (N = 75). Overall, reasons given for suicide attempts differed from reasons for nonsuicidal self-injury. Nonsuicidal acts were more often reported as intended to express anger, punish oneself, generate normal feelings, and distract oneself, whereas suicide attempts were more often reported as intended to make others better off. Almost all participants reported that both types of parasuicide were intended to relieve negative emotions. It is likely that suicidal and nonsuicidal parasuicide have multiple intents and functions. PMID- 11866175 TI - Hypnotically induced emotional numbing: a real-simulating analysis. AB - This study compared 20 real, hypnotized and 20 simulating, unhypnotized participants who were administered a hypnotic induction and then presented with emotionally distressing and neutral visual images. Half were administered a hypnotic suggestion for emotional numbing. Reals and simulators who received the emotional numbing suggestion reported comparably less responsivity to distressing stimuli than others. Whereas emotionally numb reals displayed little change in electromyographic (EMG) activity during the distressing stimuli, simulators displayed marked reduction in EMG activity. Reals (not simulators) displayed a dissociation between their affective response and awareness of the negative content of the stimuli following the numbing suggestion. PMID- 11866176 TI - Response-monitoring dysfunction in schizophrenia: an event-related brain potential study. AB - Error-monitoring abnormalities may underlie positive symptoms of schizophrenia. Response-synchronized event-related potentials during picture-word matching yielded error- and correct-response-related negativity (ERN, CRN) and positivity (Pe, Pc) and preresponse lateralized readiness potentials (LRP) from 18 schizophrenic patients and 18 controls. Both groups responded faster to matches than nonmatches, although patients were generally slower and made more errors to nonmatches. Compared with controls, patients, particularly with paranoid subtype, had smaller ERNs and larger CRNs, which were indistinguishable. LRPs showed evidence of more response conflict before errors than before correct responses in controls but not patients. Despite ERN/CRN abnormalities, post-error slowing and Pe were normal in patients, suggesting a dissociation of ERN and error awareness. Anterior cingulate and dorsolateral prefrontal cortical dysfunction in schizophrenia are implicated. PMID- 11866177 TI - Moderators of the relation between substance use level and problems: test of a self-regulation model in middle adolescence. AB - The authors tested predictions, derived from a self-regulation model, about variables moderating the relationship between level of substance use (tobacco, alcohol, and marijuana) and problems associated with use. Data were from two independent studies of adolescents, with mean ages of 15.4 and 15.5 years (Ns = 1,699 and 1,225). Factor analysis indicated correlated dimensions of control problems and conduct problems. Protective moderation was found for variables indexing good self-control; risk-enhancing moderation was found for variables indexing poor self-control. These effects were generally independent of deviance prone attitudes and externalizing symptomatology. Multiple-group structural modeling indicated moderation occurred for paths from life stress and coping motives and for paths from level to control and conduct problems. Moderation effects were also found for parental variables, peer variables, and academic competence. PMID- 11866178 TI - Lateralized perceptual organization deficits on the global-local task in schizotypal personality disorder. AB - Right and left hemisphere contributions to perceptual organization functions were examined using a divided-attention version of the global-local task in a sample of 21 unmedicated participants diagnosed with schizotypal personality disorder (SPD) and 20 controls. The SPD participants showed an abnormal global processing advantage. When the visual angle of the hierarchical stimuli was increased from 3 degrees to 9 degrees, the controls showed an increasing local processing advantage, but the SPD participants continued to show an abnormal global processing advantage. These findings suggest a local processing deficit on divided-attention versions of the global-local task in schizophrenia spectrum disorders. Female SPD participants, who had less severe interpersonal deficit symptoms, showed a more abnormal global processing advantage. Hemispheric and processing resource mechanisms that might explain these findings are discussed. PMID- 11866179 TI - Presleep attributions about arousal during sleep: nocturnal panic. AB - This study examined the role of presleep attributions about physiological events during sleep in nocturnal panic attacks. Patients who regularly experienced nocturnal panic were physiologically monitored as audio signals were presented during sleep. They were randomly assigned to 1 of 3 conditions: expected, in which signals of intense physiological changes were expected; unexpected, in which signals of intense physiological changes were not expected; or control, involving distinctly different signals unrelated to physiological responses. The unexpected condition led to substantially more self-reported distress and panic attacks. The experimental conditions did not elicit different autonomic reactions, but those who panicked showed stronger physiological responses than those who did not panic. The findings are consistent with a cognitive model of nocturnal panic attacks. PMID- 11866180 TI - Reciprocal relations between adolescent substance use and delinquency: a longitudinal latent variable analysis. AB - Reciprocal relations between self-reported substance use and delinquency were examined through the analysis of 4-wave panel data collected from 1,218 high school students. A longitudinal, latent-variable framework was used to investigate interrelationships between changing patterns of generalized involvement in these 2 problem behaviors, while simultaneously accounting for specificity in drug use (e.g., marijuana use) and delinquency (e.g., theft). Analyses revealed that a model of bidirectional effects between polysubstance use and general delinquency was plausible for boys but not for girls. For boys, the effect of delinquency on substance use was small but consistent over time, whereas the effect of substance use on delinquency was larger but restricted to the earlier waves of the study. Implications for the question of whether or not substance use and delinquency have causal connections are discussed. PMID- 11866181 TI - A longitudinal study of children's depressive symptoms, self-perceptions, and cognitive distortions about the self. AB - This longitudinal study examined how depressive symptoms relate to children's self-perceptions and to estimates of children's cognitive distortions about the self in a nonclinical sample of children who were followed from 4th grade (n = 248) through 6th grade (n = 227). Report card grades measured children's academic competence, and teachers' ratings of children's level of peer acceptance at school indicated social acceptance. Self-reported depressive symptoms predicted a change in children's negative views of the self. Moreover, the self-perceptions of children who exhibited more symptoms of depression appeared to reflect an underestimation of their actual competence. Children's negative self-perceptions and underestimations about the self were not associated with a subsequent change in depressive symptoms. The implications of the findings for cognitive theories of depression and future research with this population are discussed. PMID- 11866182 TI - Effects of nicotine dose, instructional set, and outcome expectancies on the subjective effects of smoking in the presence of a stressor. AB - The balanced placebo design (BPD) was used to evaluate the independent effects of nicotine dose and smoking-related expectancies on self-reported anxiety, urge to smoke, and withdrawal symptoms. After anxious mood was induced, participants smoked either a de-nicotinized cigarette or one with standard nicotine content. Nicotine dose was crossed with instructions that the cigarette was either de nicotinized or standard. Nicotine cigarettes produced greater anxiety reduction than de-nicotinized cigarettes. Nicotine instructions attenuated anxiety only among those who held relevant expectancies. Nicotine dose and instructional set interacted such that either nicotine cigarettes or instructions that the cigarettes contained nicotine were sufficient to reduce urge to smoke. Implications of these findings and methodological issues regarding use of the BPD with cigarettes are discussed. PMID- 11866183 TI - Clinical features of major depressive disorder in adolescents and their relatives: impact on familial aggregation, implications for phenotype definition, and specificity of transmission. AB - Three questions were addressed using family study data from a community sample: (a) Which clinical features of major depressive disorder (MDD) in adolescents are associated with elevated rates of MDD in relatives? (b) Which features of MDD in relatives distinguish family members of depressed adolescents from relatives of adolescents without mood disorders (NMD)? and (c) Do depressed adolescents with particular features have higher proportions of depressed relatives with the same features? Participants included 268 MDD adolescents, 401 NMD adolescents, and their 2,202 first-degree relatives. Rates of MDD were highest among relatives of depressed adolescents with recurrent episodes and greater impairment. Depression severity best distinguished the relatives of depressed adolescents from relatives of controls. Specific clinical features did not aggregate in families. PMID- 11866184 TI - Temperature sensitivity of the electric organ discharge waveform in Gymnotus carapo. AB - At the southern boundary of gymnotiform distribution in America. water temperature changes seasonally, and may be an environmental cue for the onset of breeding. In this study, we aim to describe the role of temperature upon electric organ discharge waveform in Gymnotus carapo, order Gymnotiformes, family Gymnotidae, and to analyze its interactions with the effects of steroid hormones. The effects of water temperature within its natural range were explored using different protocols. All fish tested had temperature-sensitive electric organ discharge waveforms: the amplitude of the last head-negative component (V4) decreased as temperature increased. Rate increases elicited by electrical stimulation had similar but smaller effect on waveform. Temperature sensitivity is a peripheral phenomenon that depends on the conductivity of the aquatic media. We found hormonal-dependent changes in the electric organ discharge waveform not previously described in this species. The amplitude and duration of V4 increased after testosterone administration. Both testosterone treatment and acclimation by sustained temperature at 27-28 degrees C (environmental simulation of breeding conditions) induced a decrease in temperature sensitivity. As in the related species Brachyhypopomus pinnicaudatus, our data strongly suggest interactions between temperature sensitivity of the electric organ discharge waveform and sexual maturity that might be crucial for reproduction. PMID- 11866185 TI - Temporal population code of concurrent vocal signals in the auditory midbrain. AB - Unique patterns of spike activity across neuron populations have been implicated in the coding of complex sensory stimuli. Delineating the patterns of neural activity in response to varying stimulus parameters and their relationships to the tuning characteristics of individual neurons is essential to ascertaining the nature of population coding within the brain. Here, we address these points in the midbrain coding of concurrent vocal signals of a sound-producing fish, the plainfin midshipman. Midshipman produce multiharmonic vocalizations which frequently overlap to produce beats. We used multivariate statistical analysis from single-unit recordings across multiple animals to assess the presence of a temporal population code. Our results show that distinct patterns of temporal activity emerge among midbrain neurons in response to concurrent signals that vary in their difference frequency. These patterns can serve to code beat difference frequencies. The patterns directly result from the differential temporal coding of difference frequency by individual neurons. Difference frequency encoding, based on temporal patterns of activity, could permit the segregation of concurrent vocal signals on time scales shorter than codes requiring averaging. Given the ubiquity across vertebrates of auditory midbrain tuning to the temporal structure of acoustic signals, a similar temporal population code is likely present in other species. PMID- 11866186 TI - Visual pigments and optical habitats of surfperch (Embiotocidae) in the California kelp forest. AB - We studied the optical microhabitat use and visual pigment variation among a group of closely related teleosts (surfperch: Embiotocidae) living along the nearshore central California coast. We employed a diver-operated spectroradiometer to record the optical microhabitat use of eight surfperch species in Monterey Bay. and microspectrophotometry to measure visual pigment absorbance for nine surfperch species. Species were dichromatic with mixtures of A1- and A2-based visual pigments exhibiting extensive maximum absorbance (lambda(max)) variation across species: 455-482 nm for SWS cones and 527-546 nm for LWS cones. Interspecific variation in sidewelling irradiance measurements (mean lambdaFmaxs) significantly accounted for 63% of the variation in surfperch LWS visual pigments and 83% of the interspecific variation in SWS visual pigments using a phylogenetically-corrected regression technique. Optimality models for maximizing relative photon capture of background radiance demonstrate that the LWS cone lambda(max) values are tuned for maximizing photon capture of the species-specific horizontal visual field, while the SWS cone lambda(max), are well offset from the dominant background radiance. This study is one of the first to demonstrate species-specific differences in habitat usage at microhabitat scales accounting for differences in photoreceptor peak absorbance among closely related, sympatric species. PMID- 11866187 TI - Satiation differentially affects performance in a learning assay by nurse and forager honey bees. AB - When not satiated prior to training, there were no differences between foragers and nurse honey bees in the acquisition of an appetitively based conditioned response in an olfactory associative learning assay, but when satiated foragers showed faster acquisition than did nurses. Satiation-related differences between foragers and nurses were more a function of behavioral state than age, because satiated precocious foragers also showed faster acquisition rates than did satiated nurse bees, despite their similar ages. Tests of sucrose responsiveness and retention of conditioned responses indicate that the observed performance differences between nurses and foragers were more likely due to differential sensitivity of sensory and motor processes related to satiation rather than differences in cognitive ability. PMID- 11866189 TI - Neurons with complex receptive fields in the stratum griseum centrale of the zebra finch (Taeniopygia guffata castanotis Gould) optic tectum. AB - The electrophysiological and morphological features of visually driven neurons of the stratum griseum centrale of the zebra finch optic tectum were studied by extracellular recording and staining techniques. Stratum griseum centrale neuron responses are sustained in most cases. Receptive fields are big, up to 150 degrees of the visual field. The excitatory center (hot spot) varies in size from 1 degrees to 15 degrees. It can be mapped by small static stimuli, adapts slower than the surround, and has a shape comparable to the excitatory fields of upper layer neurons. In contrast, the big surround shows responses only to small moving objects which elicit a typical pattern of alternating bursts and silent periods. Alternatively, the same stimuli elicit long-lasting bursts followed by strong adaption. Anatomically, stratum griseum centrale neurons are characterized by far reaching dendrites which terminate with "bottlebrush"-like endings in the upper retinorecipient layers. In addition, they are connected with retinorecipient structures by an interneuron located between layers 10 and 11. The role of the structure of inputs for the organization of the receptive fields is discussed. PMID- 11866188 TI - Temporal selectivity by single neurons in the torus semicircularis of Batrachyla antartandica (Amphibia: Leptodactylidae). AB - We investigated the response selectivities of single auditory neurons in the torus semicircularis of Batrachyla antartandica (a leptodactylid from southern Chile) to synthetic stimuli having diverse temporal structures. The advertisement call for this species is characterized by a long sequence of brief sound pulses having a dominant frequency of about 2000 Hz. We constructed five different series of synthetic stimuli in which the following acoustic parameters were systematically modified, one at a time: pulse rate, pulse duration, pulse rise time, pulse fall time, and train duration. The carrier frequency of these stimuli was fixed at the characteristic frequency of the units under study (n=44). Response patterns of TS units to these synthetic call variants revealed different degrees of selectivity for each of the temporal variables. A substantial number of neurons showed preference for pulse rates below 2 pulses s(-1), approximating the values found in natural advertisement calls. Tonic neurons generally showed preferences for long pulse durations, long rise and fall times, and long train durations. In contrast, phasic and phasic-burst neurons preferred stimuli with short duration, short rise and fall times and short train durations. PMID- 11866190 TI - Interference with the GABAergic system in the dorsolateral telencephalon and modulation of the electric organ discharge frequency in the weakly electric fish Gymnotus carapo. AB - The functional role of the dorsal portions of dorsolateral telencephalon in modulating the stable electric organ discharge was determined by microinjection of an agonist or antagonist GABAergic drug in the awake weak electric fish, Gymnotus carapo. The dorsolateral telencephalon, which is interconnected with the preglomerular complex and the dorsocentral telencephalic area was microinjected through a guide cannula previously implanted, with different doses of bicuculline, muscimol and saline. Microinjection of bicuculline into the dorsolateral telencephalon induced a complex response consisting of increase, decrease and abrupt interruptions in the frequency of electric organ discharges and an increase in motor activity. Motor activity and modulations in the electric organ discharge are dose dependent. The somatic, but not the electric, effect is abolished under anesthesia by urethane, suggesting that the two responses are parallel but unrelated in terms of occurrence. These data, together with former neuroanatomical findings by this laboratory, suggest two parallel pathways by which the blockage of GABAA receptors in the dorsolateral telencephalon causes modulations in the firing of the medullary pacemaker nucleus. A possible route for the motor effect through reticular projection from the torus semicircularis dorsalis is discussed. PMID- 11866191 TI - An evaluation of warning habits and beliefs across the adult life span. AB - Beliefs about warnings and habits associated with reading them were assessed for 863 individuals of various ages. Information gathered for various common household products included (a) how frequently people attend to warning information, (b) the degree of risk they believe is involved during product usage, and (c) how important they believe warnings are for different product types. Also assessed were perceived helpfulness and comprehension for symbols commonly found on product labels or on signs in the environment. Respondents 55 years and older reported reading product warnings more frequently than did younger adults, although they generally perceived warnings as less important. However, no overall age-related differences were found for perceived level of risk involved in using different product types. Although older adults generally perceived symbols to be very helpful when using a particular product, their comprehension levels were poorer than those of younger adults for half of the symbols. Overall, these data suggest that adults of all ages do read warnings on a variety of product types and that they believe warning information is important. This research illustrates the importance of including older adults in usability studies during the development of warning systems, given age-related effects may be associated with some aspects of the warning processing but not others. PMID- 11866192 TI - Display signaling in augmented reality: effects of cue reliability and image realism on attention allocation and trust calibration. AB - This experiment seeks to examine the relationships among three advanced technology features (presentation of target cuing, reliability of target cuing, and level of image reality and the attention and trust given to that information). The participants were 16 military personnel who piloted an unmanned air vehicle and searched for targets camouflaged in terrain, which was presented at two levels of image realism. Cuing was available for some targets, and the reliability of this information was manipulated at two levels (100% and 75%). The results showed that the presence of cuing aided target detection for expected targets but drew attention away from the presence of unexpected targets. Cuing benefits and attentional tunneling were both reduced when cuing became less reliable. Increasing image realism was compelling but increased reliance on the cuing information when those data were reliable. Potential applications include a cost-benefit analysis of how trust modulates attention in the use of automated target recognition systems and the extent to which increased realism may influence this trust. PMID- 11866193 TI - Monitoring task loading with multivariate EEG measures during complex forms of human-computer interaction. AB - Electroencephalographic (EEG) recordings were made while 16 participants performed versions of a personal-computer-based flight simulation task of low, moderate, or high difficulty. As task difficulty increased, frontal midline theta EEG activity increased and alpha band activity decreased. A participant-specific function that combined multiple EEG features to create a single load index was derived from a sample of each participant's data and then applied to new test data from that participant. Index values were computed for every 4 s of task data. Across participants, mean task load index values increased systematically with increasing task difficulty and differed significantly between the different task versions. Actual or potential applications of this research include the use of multivariate EEG-based methods to monitor task loading during naturalistic computer-based work. PMID- 11866194 TI - Effect of flooring on standing comfort and fatigue. AB - This study investigated the influence of flooring on subjective discomfort and fatigue during standing and on potentially related objective measures. Participants stood for 4 h on each of 7 flooring conditions while performing computer tasks. During the 3rd and 4th h, floor type had a significant effect on a number of subjective ratings, including lower-leg and lower-back discomfort/fatigue and 2 of 4 objective variables (center of pressure weight shift and lower-extremity skin temperature). In addition, lower-leg volumetric measurements showed tendencies toward greater lower-extremity swelling on uncomfortable floors. The hard floor and 1 floor mat condition consistently had the worst discomfort/fatigue and objective ratings. Significant relationships were noted between the affected subjective ratings and objective variables. In general, floor mats characterized by increased elasticity, decreased energy absorption, and increased stiffness resulted in less discomfort and fatigue. Thus flooring properties do affect low-back and lower-leg discomfort/fatigue, but the result may be detectable only after 3 h of standing. Potential applications of this research include the reduction of work-related health problems associated with long-term standing. PMID- 11866195 TI - Flying personal planes: modeling the airport choices of general aviation pilots using stated preference methodology. AB - This study employed stated preference (SP) models to determine why general aviation pilots choose to base and operate their aircraft at some airports and not others. Thirteen decision variables identified in pilot focus groups and in the general aviation literature were incorporated into a series of hypothetical choice tasks or scenarios. The scenarios were offered within a fractional factorial design to establish orthogonality and to preclude dominance in any combination of variables. Data from 113 pilots were analyzed for individual differences across pilots using conditional logit regression with and without controls. The results demonstrate that some airport attributes (e.g., full-range hospitality services, paved parallel taxiway, and specific types of runway lighting and landing aids) increase pilot utility. Heavy airport congestion and airport landing fees, on the other hand, decrease pilot utility. The importance of SP methodology as a vehicle for modeling choice behavior and as an input into the planning and prioritization process is discussed. Actual or potential applications include the development of structured decision-making instruments in the behavioral sciences and in human service programs. PMID- 11866196 TI - Effect of geometric field of view on stereoscopic spatial judgments. AB - Within a stereoscopic display the field of view (FOV) was held constant at 13.86 degrees while the geometric field of view (GFOV) was varied across four levels: 0 degrees (parallel), 13.86 degrees (veridical), 50 degrees and 100 degrees. Participants performed a distance-matching task where they adjusted the distance of a standard track from the centre of the display to match the distance of a target track from the same point. The results indicated that while the least error occurred in the veridical GFOV condition, small variations of GFOV away from the veridical have little effect. Large differences between FOV and GFOV (36 degrees and 86 degrees) increased errors markedly. A trend toward better performance in the veridical GFOV condition relative to the parallel GFOV condition suggests that the use of linear perspective information in a stereoscopic display may facilitate more accurate spatial perception. Actual or potential applications of this work include stereoscopic display design in aviation and non-aviation settings. PMID- 11866197 TI - Configural display design techniques considered at multiple levels of evaluation. AB - Two studies were conducted to examine issues in the design and evaluation of configural displays. Four design techniques (bar graphs/extenders, scale markers/ scale grids, color coding/color layering/color separation, and annotation with digital values) were applied, alone and in combination, to a baseline configural display, forming 10 displays. Two qualitatively different evaluations assessed performance for (A) low-level data probes (quantitative estimates of individual variables) and (B) system control and fault detection tasks. Three of the four design techniques improved performance significantly for low-level data probes (color coding was the exception). A display with digital values only (i.e., no analog configural display) produced the poorest performance for control/fault detection tasks. When both levels of evaluation are considered, a composite display (configural display with all four techniques applied) was clearly the most effective. Overall, the findings obtained in the two experiments provide very limited evidence for the generalization of results between evaluations. The two levels of evaluation, the display manipulations, and the patterns of results are considered in terms of a cognitive systems engineering evaluation framework. General implications for the evaluation of displays and interfaces are discussed. Actual or potential applications include design techniques to improve graphical displays and methodological insights to focus and improve evaluation efforts. PMID- 11866198 TI - Cursor orientation and computer screen positioning movements. AB - Implicit directional cues in arrowhead cursors could influence positioning of a cursor on the screen of the computer. Performance during cursor placement may benefit from compatibilities between cursor orientation and direction of movement. Arrowheads could also elicit illusory processes that may affect judgments of (a) the distances on the screen or (b) the location of the point of the arrowhead. To address the impact of the cursor's orientation on its positioning, we had 12 participants move cursors (crosshairs, leftward, or rightward arrow) leftward or rightward to targets (near, far) on a computer screen. Movement amplitude was more important than cursor orientation for initiation of rightward movements, whereas cursor orientation affected the duration of leftward movements and movements to farther targets. Arrowhead orientation contributed to the greater overshooting of far targets. There was little evidence that compatibility of orientation and direction of movement assisted response initiation or execution, and there was little indication that arrowhead cursors led to illusory effects that influenced cursor placement. Arrowhead cursors can provide irrelevant stimulus dimensions that distract users. This work can be applied to the design of cursors in graphical user interfaces. The use of orientation-neutral cursors or cursors whose stimulus dimensions are more relevant is recommended. PMID- 11866199 TI - Distance perception of vehicle rear lights in fog. AB - Perceptual difficulty is one of the main explanations given for the behavioral modifications and high accident rate associated with driving in fog. The present study investigates how fog and the characteristics of vehicle rear lights affect distance perception. Two experiments in a fog chamber (meteorological visibility range of 5-15 m) were run under conditions simulating nighttime fog. The participants gave verbal estimates of the distance (8-28 m) of vehicles simulated by rear-light arrangements. The results revealed an average increase of 60% in the perceived distance of vehicles in fog as compared with normal visibility conditions. Distance overestimation was particularly pronounced when the vehicle had only 1 fog light instead of 2 and when the lights were close together. No effect of light height was observed. These results suggest that the perception of vehicle distance in nighttime fog could be significantly improved by the presence of 2 lights with maximal spacing. Actual or potential applications of this research include vehicle rear light design. PMID- 11866200 TI - Effects of navigation speed on motion sickness caused by an immersive virtual environment. AB - This study investigated the effects of navigation speed on the level of motion sickness during and after a 30-min head-steered virtual environment. Root-mean squares for 8 speeds in the fore-and-aft axis were 3, 4, 6, 8, 10, 24, 30, and 59 m/s. Participants were 96 Chinese men. Both the nausea and vection ratings increased significantly with speeds increasing from 3 m/s to 10 m/s. At speeds exceeding 10 m/s, the ratings stabilized. Navigation speeds were found to significantly affect the onset times of vection and nausea but did not affect their rates of increase with duration of exposure. For the various Simulator Sickness Questionnaire scores, navigation speed had a significant influence on only the oculomotor subscore. Actual or potential applications of this research include the prediction of sickness associated with simulation tours in a virtual environment at different navigation speeds. PMID- 11866201 TI - Human performance models and rear-end collision avoidance algorithms. AB - Collision warning systems offer a promising approach to mitigate rear-end collisions, but substantial uncertainty exists regarding the joint performance of the driver and the collision warning algorithms. A simple deterministic model of driver performance was used to examine kinematics-based and perceptual-based rear end collision avoidance algorithms over a range of collision situations, algorithm parameters, and assumptions regarding driver performance. The results show that the assumptions concerning driver reaction times have important consequences for algorithm performance, with underestimates dramatically undermining the safety benefit of the warning. Additionally, under some circumstances, when drivers rely on the warning algorithms, larger headways can result in more severe collisions. This reflects the nonlinear interaction among the collision situation, the algorithm, and driver response that should not be attributed to the complexities of driver behavior but to the kinematics of the situation. Comparisons made with experimental data demonstrate that a simple human performance model can capture important elements of system performance and complement expensive human-in-the-loop experiments. Actual or potential applications of this research include selection of an appropriate algorithm, more accurate specification of algorithm parameters, and guidance for future experiments. PMID- 11866202 TI - Role of lateral acceleration in curve driving: driver model and experiments on a real vehicle and a driving simulator. AB - Experimental studies show that automobile drivers adjust their speed in curves so that maximum vehicle lateral accelerations decrease at high speeds. This pattern of lateral accelerations is described by a new driver model, assuming drivers control a variable safety margin of perceived lateral acceleration according to their anticipated steering deviations. Compared with a minimum time-to-lane crossing (H. Godthelp, 1986) speed modulation strategy, this model, based on nonvisual cues, predicts that extreme values of lateral acceleration in curves decrease quadratically with speed, in accordance with experimental data obtained in a vehicle driven on a test track and in a motion-based driving simulator. Variations of model parameters can characterize "normal" or "fast" driving styles on the test track. On the simulator, it was found that the upper limits of lateral acceleration decreased less steeply when the motion cuing system was deactivated, although drivers maintained a consistent driving style. This is interpreted per the model as an underestimation of curvilinear speed due to the lack of inertial stimuli. Actual or potential applications of this research include a method to assess driving simulators as well as to identify driving styles for on-board driver aid systems. PMID- 11866203 TI - What determines whether observers recognize targeted behaviors in modeling displays? AB - Observational learning is based on a critical assumption that trainees can and do recognize critical modeled behaviors. This assumption has been virtually untested in applied settings. We studied the effects of work experience and instructions on the ability of 59 observers to recognize target behaviors in an observational learning paradigm similar to existing ones. Additionally, we investigated the effects of two key factors that were hypothesized to affect the recognition process in observational learning. The results indicated that only observers who had a minimum of work experience (i.e., intermediate and experienced observers in the study) were able to consistently recognize targeted behaviors. Additionally, recognition was influenced by the level of detail of instructions given to the participants. Finally, characteristics of the modeled behaviors greatly affected recognition: Overall, examples of negative behaviors were better recognized than were positive examples. Behaviors whose consequence was shown were also better recognized than those that were neither reinforced nor punished in the video. The results are discussed in terms of their implications for the design of observational learning as a training strategy in complex and applied social learning situations. The applications of this work include the design of training, and the training of evaluators and observers. PMID- 11866204 TI - Long-term follow-up and prognostic significance of angiogenic basic fibroblast growth factor (bFGF) expression in patients with breast cancer. AB - The development of distant metastasis in breast cancer patients is the key step towards worse prognosis. The angiogenic basic fibroblast growth factor (bFGF) has been associated with tumorigenesis and metastasis in several human cancers. Therefore, bFGF expression was studied by immunohistochemistry in 111 patients with primary breast cancer. The results were correlated with prognostically relevant clinico-pathological features. such as tumor stage, grading. nodal stage and survival. bFGF was expressed in approximately 70% of the breast cancer tissues; 30% of the tumors showed strongly positive staining. With the exception of histological grading (p < 0.05), no correlation was found between the extent of bFGF expression and prognostic parameters. Analysis of survival showed a significantly (p < 0.05) prolonged survival for patients with a concomitant absence of axillary lymph node metastasis and bFGF immunoreactivity. Our data suggest that increased bFGF expression is a novel parameter for worse prognosis in nodal-negative breast cancer patients. PMID- 11866205 TI - DNA ploidy and S-phase fraction analyses of hyperplastic, atypical and cancerous endometrium using flow cytometry from paraffin-embedded tissues. AB - Atypical hyperplasia of the endometrium is an entity that needs to be clearly distinguished from other florid hyperplastic states on the one hand and from well differentiated adenocarcinomas on the other. This may at times be difficult on pure morphological grounds. In this study, DNA ploidy and S-phase fraction of hyperplastic, atypical and cancerous endometrium were evaluated using flow cytometry from paraffin-embedded tissues. In total, 72 cases (24 hyperplastic, 24 atypical and 24 adenocarcinomas) were selected. All hyperplastic endometria showed a diploid stemline, while 2/24 atypical hyperplasias showed aneuploid (Near-diploid) peaks. Both of these cases were severely atypical and one of these, on hysterectomy, showed early invasive carcinoma. There was no significant difference in mean S-phase fractions of hyperplastic vs. atypical endometria. DNA aneuploidy was significantly more common with much higher S-phase fractions in poorly and moderately differentiated carcinomas than in well-differentiated ones. It was concluded that aneuploid (near-diploid) peaks, if ever present in atypical hyperplasias, may indicate an aggressive disease/neoplastic transformation. PMID- 11866206 TI - Chlamydia pneumoniae in different coronary artery segments in the young. AB - Chlamydia pneumoniae has emerged as the most likely pathogen to have a causative role in the development and/or for progression of atherosclerosis. Evidence for this is based on epidemiological and pathological studies. In an effort to better understand the significance of finding C. pneumoniae in atheromata, we examined coronary artery segments of young adults (15-34 years) with and without atherosclerosis. Left anterior descending coronary arteries (LAD) of 74 young adults who died suddenly were examined histologically and for the presence of C. pneumoniae by immunohistochemistry. C. pneumoniae was identified in advanced lesions (Stary types III to VI) in 17 of 32 cases (53%), and in early lesions (Stary type I-II) in 8 of 37 cases (21%), mainly at the proximal segments of the LAD. C. pneumoniae was not found in the intimal and medial layer of normal appearing coronary arteries. C. pneumoniae was detected in the adventitia in 51 (67%) coronary arteries: in 27 of normal arteries and early lesions (64%), and in 24 of atherosclerotic lesions (75%). C. pneumoniae was found most often in macrophages, less offen in smooth muscle cells. We also observed a correlation between C. pneumoniae positivity and cigarette smoking. IN CONCLUSION: C. pneumoniae may relate to the severity of atherosclerosis in young people, and it may thus initiate atherosclerotic injury or facilitate its progression with other risk factors. PMID- 11866207 TI - Pathologic findings in the adrenal glands of autopsied patients with acquired immunodeficiency syndrome. AB - A morphologic evaluation was carried out on adrenal glands from 128 autopsied patients with the acquired immunodeficiency syndrome (AIDS). The adrenal gland was compromised in 99.2% of the cases, with distinct pathological features and infectious agents. Inflammatory infiltrates were observed in 99.2% of the cases with a predominance of mononuclear cells in 97.4%, affecting mainly the medulla. Necrosis, fibrosis, hemorrhages and neoplasias were observed. We also described 3 (2.3%) cases of calcification located in the adrenal gland central vein (AGCV). This is seldom mentioned in the literature. Cytomegalovirus was the most frequent infectious agent, observed in 48.4% of cases. Balamuthia mandrillaris, a free living ameba, was found in one case affecting the entire gland. We also found a nest of Trypanosoma cruzi in the musculature of the AGCV. The presence of the nest of T cruzi in AGCV may play a role in the reactivation of this infection in immunosuppressed individuals. Pathologic processes and opportunistic infections may contribute to the alterations in the adrenal gland that lead to multiple organ failure observed in terminal AIDS patients. PMID- 11866208 TI - Tuberculous otitis media: a difficult diagnosis and report of four cases. AB - Tuberculous otitis media is a rare disease. Due to the condition's rarity and its usually indolent course, the diagnosis is often delayed. This can lead to irreversible complications, such as permanent hearing loss or facial nerve paralysis. Tuberculosis of the middle ear cleft, as this disease's first presentation, is indeed very rare. Surgery may be carried out prior to diagnosis occasionally, i.e., middle ear exploration for chronic middle ear disease. We present four cases of tuberculous otitis media which occurred as the first presentation of the disease. The patients did not present with the classic symptoms of middle ear tuberculosis. The diagnosis was based on the histology following middle ear exploration for chronic middle ear disease. None of the patients presented any other systemic involvement. We present a review of this disease's clinical symptoms and the diagnostic tests available. PMID- 11866209 TI - Intrarenal solitary fibrous tumor of the kidney report of a case with emphasis on the differential diagnosis in the wide spectrum of monomorphous spindle cell tumors of the kidney. AB - Solitary fibrous tumor (SFT) is a neoplasm that can occur in the urogenital tract, and is also reported occurring in the spermatic cord, seminal vesicles, urinary bladder, prostate, and kidney. Furthermore, it is most important to consider its existence in the kidney, because it is usually diagnosed as renal cell carcinoma pre-operatively. To our knowledge, only 10 cases of SFT have been reported in the kidney to date. We report the clinico-pathological features of an intrarenal SFT occurring in a 31-year-old woman. The tumor, measuring 8.6 cm in its greatest diameter, completely replaced the cortex and the medulla of the middle region of the right kidney, compressing the pelvis. Radiological imaging was consistent with a renal cell carcinoma. Histologically, the tumor was composed of a proliferation of bland-looking vimentin+, CD34+, bcl2+ and CD99+ spindle cells exhibiting a haphazard to storiform growth pattern, pushing borders, and a low mitotic rate (2 mitoses x 10 HPF). We placed emphasis on the differential diagnostic problems, i.e., its differentiation from other primary monomorphous benign and malignant spindle cell tumors of the kidney, such as fibroma, benign fibrous histiocytoma, hemangiopericytoma, inflammatory myofibroblastic (pseudo-)tumor, leiomyoma, angiomyolipoma with predominant spindle cell smooth muscle component, benign peripheral nerve sheath tumors, renal mixed epithelial/stromal tumors, adult type mesoblastic nephroma, fibrous type monophasic synovial sarcoma, malignant peripheral nerve sheath tumors, fibrosarcoma, and low-grade fibromyxoid sarcoma. PMID- 11866210 TI - Extensive plasma cell infiltration with crystal IgG inclusions and mutated IgV(H) gene in an osteoarthritis patient with lymphoplasmacellular synovitis. A case report. AB - The presence of immunoglobulin crystal inclusions in plasma cells from plasmacytomas and B-NHLs (linked to overstimulation and overproduction) has been frequently reported. Our case describes a lymphoplasmacellular synovitis in a patient with osteoarthritis (OA) showing an unusually high plasma cell infiltration and for the first time crystals in plasma cells. Using immunohistochemistry. these crystals were identified as being IgG with a balanced lambda/kappa ratio. IgV(H) gene analysis (n = 5 clones) showed that they were somatically mutated (R/S of CDR > 3): in one case, an insertion of 9 nucleotides on the CDR2 region was observed. High R/S values in the CDR indicated antigen selectivity and affinity (4/5). Since no germinal centers could be detected and the analyzed B cells showed antigen selectivity, it may be concluded that already antigenically activated B cells migrated into the synovium and locally differentiated into plasma cells, leading to the extensive infiltration observed. Rheumatoid fibroblasts were shown to support terminal B cell differentiation. Our data suggests that the ability of fibroblasts to activate B cells is not only restricted to RA, but also occurs in OA. The intense plasma cell infiltration contributed to further cartilage damage by altering the microenvironment of the nourishing synovial tissue or by the local production of pathogenic autoantibodies. PMID- 11866211 TI - Ancient schwannoma arising in a lymph node: report of a case and review of the literature. AB - We report a case of an intranodal schwannoma in a 79-year-old woman. The patient underwent a left colectomy for a colonic adenocarcinoma, with regional lymph node dissection. Macroscopic examination of the specimen revealed a well-circumscribed 4 cm nodule located in the pericolic fat. Macroscopically, it was interpreted as a metastatic lymph node. Microscopically, the nodule was composed of a proliferation of bland spindle cells, was immunohistochemically positive for S100 protein, and negative for smooth muscle actin, desmin, and cytokeratin. The pathological findings led to the diagnosis of a very rare case of primary schwannoma of the lymph node. PMID- 11866212 TI - Complex pathological diagnosis of granulocytic sarcoma: apropos of a case. AB - The diagnosis of granulocytic sarcoma can be very difficult when there is no demonstrable abnormality in the peripheral blood or bone marrow. We present the diagnostic algorithm of granulocytic sarcoma by reporting on a case mimicking large cell lymphoma without previous manifestation of acute myeloid leukemia or a myeloproliferative disorder. After standard histoprocessing, we used immunohistochemical and molecular biological methods to analyze our case. The lymph node showed diffuse infiltration of immature blast cells resembling large cell lymphoma. However, immunohistochemistry did not support this diagnosis. The tumor cells showed LCA, bcl-2, CD43, CD34 and myeloperoxidase positivity. We also detected bcl-2 gene rearrangement. In case of a lack of a specific histological picture, particularly in poorly differentiated tumors, only some minor histological signs in combination with immunohistochemistry and molecular diagnostic methods can help to render the correct diagnosis. PMID- 11866213 TI - Brain heterotopia in pharyngeal region. A morphological and immunohistochemical study. AB - Pharyngeal brain heterotopia is a congenital and generally biologically benign lesion. In contrast to brain heterotopia in the nose, the most common site of this lesion, brain heterotopia in the pharynx is very rare. Pharyngeal heterotopic tissue can be composed of various components, i.e., astrocytes, neurons, ependyma or choroid plexus, oligodendrocytes, retina, and, occasionally, neoplastic nodules. In contrast, nasal lesions are often only composed of astrocytes. We report a case of brain heterotopia in the pharyngeal region, diagnosed in a newborn female infant, causing serious respiratory distress. The infant underwent surgical excision of the lesion, and after 1 year of follow-up, she is recurrence-free. The mass, about 3 cm in diameter and showing no connection with encephalic structures, was characterized by numerous papillary structures and areas containing stellate-like or spindle cells focally forming nodules. Moreover, there was inflammatory infiltration, whereas mitoses, hemorrhages, and necroses were absent. Immunohistochemistry revealed a choroid plexus nature of the papillary formations (S-100, cytokeratins, transthyretin and vimentin-positive) and the presence of glial and neuronal cells in the remaining areas (glial fibrillary acidic protein, neuron-specific enolase, neurofilaments, synaptophysin, and S-100 positive). This case report confirms that the presence of choroid plexus is not uncommon and that pharyngeal brain heterotopia is usually benign. PMID- 11866214 TI - Acute carpal tunnel syndrome due to filarial infection. AB - A case of acute carpal tunnel syndrome (CTS) due to filarial infection is reported in a resident of Newfoundland, Canada. The patient presented with classical symptoms and signs of acute CTS and a filarial worm was identified in synovial tissue removed during surgery that was morphologically consistent with the genus Brugia. Antifilarial antibodies were present in serum. Surgical removal of the infected tissue, without administration of antifilarial drugs, proved adequate in eradicating the infection and a subsequent serum sample was negative for antifilarial antibodies. The patient remains asymptomatic more than two years after surgery. This case is unusual in its clinical presentation and geographic location. PMID- 11866215 TI - Prognostic importance of tumor angiogenesis in breast carcinoma with adjuvant chemotherapy. AB - Tumor angiogenesis is believed to be related to prognostic factors involved in tumor development and metastasis. Using immunohistochemical methods, we evaluated tumor angiogenesis in 42 early invasive breast cancer patients (T1-2, NO-1-2, M0). Four patients received tamoxifen, 25 patients received CAF or CA, and 15 patients received CMF as adjuvant therapy. The median follow-up was 47 (range 24 119) months. Ten patients (43.5%) in the node-positive group and 2 patients (10.5%) in the node-negative group relapsed (p = 0.019). The mean microvessel count (MVC) was 60.3 3.05 per 200x field (range: 16-95). MVCs of postmenopausal and premenopausal patients were 50.13 +/- 5.74 and 68.64 +/- 4.11, respectively, in the axillary lymph node (ALN)-negative patient group (p = 0.04). Staining was moderate to strong in 13 (68%) ALN-negative and in 17 (74%) ALN-positive patients (p > 0.05), and was also moderate to strong in 82% of premenopausal patients and in 50% of postmenopausal patients (p = 0.037). There was no significant relationship between angiogenesis and p53, nor was angiogenesis significantly associated with the patient ER status and tumor size. No significant correlations were found between OS/DFS and Factor VIII staining or p53 (log rank test, p > 0.05). Of all ALN-negative patients with increased angiogenesis, one patient of the CMF group relapsed, but no recurrence occurred in patients undergoing anthracycline-based chemotherapy (p > 0.05). On the other hand, of all ALN positive patients with increased angiogenesis, 5/14 patients treated with anthracylcine and 2/2 CMF-treated patients relapsed (p = 0.175). Despite the statistical insignificance, anthracycline-based adjuvant chemotherapy appears to be more effective than CMF as regards relapse prevention particularly in early ALN-positive breast cancer patients with increased angiogenesis. Additional studies are necessary to demonstrate the clinical importance of angiogenesis. PMID- 11866216 TI - Cirrhosis and chronic hepatitis. PMID- 11866217 TI - MR contrast agents in hepatic cirrhosis and chronic hepatitis. AB - Chronic liver disease alters the gross architecture of the liver and its arterial and portal blood supply. The relative proportion of regenerative hepatocytes, necrosis, extracellular interstitial space, and fibrosis is responsible for liver enhancement after the administration of a contrast agent. Because contrast agents can be directed toward either the extracellular or the intracellular spaces, knowledge of the different parenchymal enhancement alterations seen after the administration of these agents is necessary to understand how chronic liver disease pathologic changes influence contrast-enhanced magnetic resonance (MR) images. This article reviews the effect of chronic liver disease on MR contrast enhancement, as well as the effect of altered enhancement on lesion detection and characterization. Both extracellular and intracellular contrast agents are considered. PMID- 11866218 TI - Radiology in liver transplantation. AB - Imaging studies are becoming essential in the management of orthotopic liver transplantation (LT). They have a very important role in the preoperative evaluation and selection of suitable candidates. At the same time, they are essential in the early detection of postoperative complications, the recognition of which allows the prompt institution of appropriate therapeutic measures. Timely recognition of complications improves the success of LT; furthermore, some complications can be treated with interventional radiologic procedures, avoiding additional surgery. This article reviews the current application of diagnostic and interventional imaging in liver transplantation, both for cadaveric and living donor transplants. PMID- 11866219 TI - Vascular imaging and interventional procedures in hepatic cirrhosis. AB - Changes in the vascularization of the cirrhotic liver are related to the progression of the disease. Knowledge of normal hepatic vascular anatomy and anatomic is essential for understanding the altered hepatic circulation seen in cirrhosis. We analyze the changes in liver perfusion with special interest in the anatomic features that are important in interventional procedures. The indications, technical notes, and complications of transjugular liver biopsy, transiugular intrahepatic portosystemic shunt (TIPS), and embolization of hepatocellular carcinoma, are reviewed. PMID- 11866220 TI - Doppler in hepatic cirrhosis and chronic hepatitis. AB - Doppler ultrasound and color Doppler are being used routinely in the study of vascular structures of the abdomen, and more particularly the liver. Reports published in the literature to date have shown that all patients with hepatic cirrhosis and chronic hepatitis should be studied in the first stage of their illness and in follow-up by using Doppler techniques. With new ultrasound software and the latest generation of contrast agents, we can obtain a better and more complete vascular study of the liver, which previously could be achieved only with triple-phase CT. Therefore, hepatic cirrhosis and chronic hepatitis, as well as their vascular complications, can be followed up more closely. The vascular complications include flow alterations in the hepatic artery and veins, portal hypertension, portal vein thrombosis, portosystemic shunts, and vascularization associated with liver tumors. PMID- 11866221 TI - Gray-scale ultrasound in hepatic cirrhosis and chronic hepatitis: diagnosis, screening, and intervention. AB - Chronic hepatitis and hepatic cirrhosis are pathologies with high prevalence in the world population. Ultrasound (US) allows for a quick and precise examination of the liver parenchyma, the vascular structures, the biliary tract, and the abdominal cavity. Changes can be detected in the pattern of liver echostructure that suggest the presence of chronic liver disease, portal hypertension, and the presence of liver tumors. Moreover, US guidance provides an easy way for performing interventional procedures, such as biopsies for classifying the degree and type of liver disease, biopsies of focal liver lesions, and the application of percutaneous treatments for hepatocellular carcinoma (HCC). In this article we discuss the multiple applications of US in the management of patients with chronic liver disease. PMID- 11866222 TI - CT in hepatic cirrhosis and chronic hepatitis. AB - Cirrhosis is a diffuse liver disease with premalignant potential in which hepatocellular carcinoma (HCC) frequently develops. The hemodynamics of contrast material are the key to diagnosis of focal liver lesions with computed tomography (CT). Lesions with arterial-dominant vascularity, such as HCC, show brisk enhancement during the arterial phase, whereas lesions with portal blood supply can appear as hyperenhancing lesions in the portal phase. The advent of helical CT has significantly improved the CT examination of the liver because the arterial phase can be displayed independently of the portal phase. The addition of arterial phase imaging to conventional portal phase imaging seems to improve tumor detection and characterization. Although HCC is the single most frequent tumor seen in chronic liver disease, other lesions such as peripheral cholangiocarcinoma and hemangioma should be considered in the differential diagnosis. Optimization of helical CT techniques may allow better detection and characterization of these lesions. In addition to tumor detection, CT plays an important role in preoperative staging of HCC as well as in preoperative assessment of patient candidates to hepatic transplantation. The use of CT angiography with maximum intensity projection techniques may allow for better preoperative work-up and vascular mapping in HCC patients. This article shows the spectrum of helical CT findings in chronic liver disease and specifically in the imaging of HCC and other focal lesions. PMID- 11866224 TI - MR imaging in chronic hepatitis and cirrhosis. AB - The role of magnetic resonance imaging (MRI) in the evaluation of diffuse parenchymal abnormalities of the liver has been expanded by recent technical advances of MR systems as well as the evolution of intravenous contrast media. Currently, MR is undoubtedly the most useful imaging modality for detecting the presence of chronic liver disease. Tailored sequences allow acurate depiction of specific disorders, including steatohepatitis and iron-overload states. Morphologic changes and signal intensity effects not only facilitate the diagnosis of chronic liver disease with MRI but they also help to distinguish between different etiologies, and they assist in staging the histologic severity of certain chronic conditions. Moreover, the faster MRI scanning techniques presently available permit the dynamic assessment of contrast enhancement, which permits improved characterization of focal hepatic lesions, including regenerative nodules, dysplastic nodules, and hepatocellular carcinoma (HCC). Although overlap in MRI findings still may exist among different types of chronic liver disease and among focal liver lesions, familiarity with certain specific imaging features may be diagnostic in the proper clinical setting. Finally, comprehensive MRI examination, including MR angiography and MR cholangiography, is the most sensitive and cost-effective technique for detecting extrahepatic disease, diagnosing vascular disorders, and evaluating the patient before or after liver transplantation. This article focuses on the current role of MR imaging in patients with chronic liver disease. The subjects covered include the detection and characterization of chronic hepatitis and cirrhosis, specific findings seen in steatohepatitis and certain metabolic diseases, the evaluation of extrahepatic vascular complications of cirrhosis, and patient assessment before and after liver transplantation. The characterization of hepatic masses is also included briefly. This subject is covered in greater depth elsewhere in this issue. PMID- 11866223 TI - Magnetic resonance of focal liver lesions in hepatic cirrhosis and chronic hepatitis. AB - Detection of focal liver nodules in patients with cirrhosis continues to be a radiologic challenge despite progressive advances in liver imaging in the past 2 decades. Patients with hepatic cirrhosis have a high predisposition to develop hepatocellular carcinoma (HCC), and the early detection and diagnosis of this tumor is very important because the most effective treatment is surgical resection, transplantation, or local ablation therapy when the tumor is small. Cirrhotic livers are mainly composed of fibrosis, together with a broad spectrum of focal nodular lesions ranging from regenerative nodules to premalignant dysplastic nodules to overt HCC. Awareness of such lesions and interpretation of imaging studies in these patients requires a critical review to detect subtle tumors, and a thorough understanding of the imaging appearance of the malignant and benign masses that can occur in the cirrhotic liver. Although the recent advances in liver imaging techniques, especially computed tomography (CT) and magnetic resonance (MR), have facilitated the detection and characterization of focal liver nodules in cirrhotic patients, discriminating between HCC and precancerous nodules remains problematic with all available imaging techniques. Nevertheless, MR imaging appears to have more potential than other imaging techniques in the study of cirrhotic patients and MR may be more appropriate than the other imaging modalities for the detection of small HCCs. In this article we review the imaging characteristics of nodular focal lesions that arise in cirrhotic livers, with special attention to MR imaging features. PMID- 11866225 TI - Pharmacologic management of childhood hypertension: current status, future challenges. AB - The treatment of childhood hypertension has been hampered by several factors, including a lack of extensive scientific data regarding drug efficacy and pharmacokinetics in children, a lack of manufacturer's recommendations for the use of antihypertensive agents in children, and a lack of age-appropriate drug formulations. These problems have forced clinicians either to rely on limited data from older studies of agents empirically no longer considered first-line choices for use in children, or to adapt agents studied in adults for pediatric use. Recent developments, including publication of single-center studies of newer agents and the proliferation of industry-sponsored trials spurred by the Food and Drug Administration Modernization Act, have increased the amount of data available. Investigators involved in such studies must ensure that the unique aspects of using antihypertensive drugs in children are adequately addressed. PMID- 11866226 TI - Antihypertensive therapy: basic pharmacokinetic and pharmacodynamic principles as applied to infants and children. AB - Increased attention has been paid to the study of antihypertensive agents in children following the Food and Drug Administration Modernization Act. Many short term studies of drug effectiveness and safety are underway or recently completed. Defining the disposition of antihypertensive agents in the pediatric population has become an important part of ongoing efforts to label antihypertensive agents for use in children. In comparison with larger trials done in the adult population, relatively few pediatric subjects will be studied. Hence, it is important to gather as much information as possible for each agent during pediatric trials. Appropriate study design is dependent not only on a working knowledge of the unique features of hypertension in children, but also on a practical assessment of factors that affect drug disposition in children. Basic principles of pharmacokinetics (PK), pharmacodynamics (PD), and PK-PD modeling as applied to pediatric antihypertensive therapy are briefly considered. PMID- 11866227 TI - Ambulatory blood pressure monitoring in clinical trials in adults and children. AB - Ambulatory blood pressure (BP) monitoring is a technique associated with several operational characteristics that make its use in adult hypertension trials attractive. To extend its use effectively to hypertension trials in children, additional studies are needed to assess its performance in this population. It is likely that ambulatory monitoring will be important in this population for the study of drug efficacy and safety. In general, ambulatory monitoring has better short-term and long-term reproducibility than office BP. Some data suggest that this superiority of ambulatory over office BP is also true in children. This better reproducibility should allow a drug effect to be detected with a smaller sample size, especially in crossover studies. Ambulatory BP averages are also free from the office-induced increase in BP, making it is easier to include adults and children in trials who have ambulatory hypertension. Such a requisite for a trial has to be planned to include more patients in the recruitment and run in phases as use of an ambulatory criterion for hypertension will lead to some patients with elevated office BP not qualifying based on ambulatory readings. Of additional major importance, ambulatory monitoring of BP correctly identifies the temporal effects of a drug and hence allows accurate calculation of its trough-to peak ratio. Ambulatory monitoring promises to be a significant tool for the study of hypertension in children including pathophysiology and the efficacy of antihypertensive drugs. PMID- 11866228 TI - Birth weight as a predictor of future hypertension. AB - According to the concept of "fetal programming," intrauterine conditions that impair fetal growth can program the fetus to express hypertension, as well as other chronic diseases, in later life. A model for the development of hypertension that includes the effect of the intrauterine environment on fetal growth as a mechanistic pathway contributes a new dimension in causality and is of great interest. Experimental studies that apply maneuvers to impose fetal undernutrition in animal models have developed data that support this concept. The evidence developed from epidemiologic investigations on humans is less conclusive. Overall, there tends to be a small but consistent relationship of lower birth weight with higher blood pressure (BP). For each kilogram increase in birth weight, the systolic BP is 1 to 2 mm Hg lower. PMID- 11866229 TI - Perinatal programming, renal development, and adult renal function. AB - Substantial data indicate that maternal diet during gestation, as well as perinatal events, affect organogenesis and, furthermore, may affect organ function at maturity, thus determining whether an individual is prone or resistant to a variety of health problems such as hypertension, cardiovascular disease, or renal dysfunction. This concept is called perinatal programming, or the fetal origins of adult disease (FOAD), a concept that arose after the publication of epidemiologic observations by Barker and colleagues that birth weight among apparently normal individuals studied in midlife was inversely associated with the presence of cardiovascular disease and hypertension. We now review evidence from studies in experimental animal models that have used nutritional and pharmacologic manipulation to extend epidemiologic observations. PMID- 11866230 TI - Obesity hypertension. AB - The association between obesity and hypertension is well recognized. However, the exact mechanisms whereby obesity causes hypertension are complex and multifactorial. The current article summarizes some of the known mechanisms responsible for obesity hypertension. PMID- 11866231 TI - Effect of hypertension on the progression of chronic renal failure in children. AB - This article reviews the current state of knowledge concerning the vicious cycle of hypertension and progressive loss of renal function in renal disease, as well as the renoprotective potential of antihypertensive treatment, with a specific focus on children and adolescents. Deficient arteriolar autoregulation renders damaged kidneys particularly sensitive to systemic high blood pressure (BP). Intraglomerular hypertension promotes proteinuria, which further activates the renin-angiotensin system (RAS). Angiotensin II, apart from its vasoconstrictor effects, induces local proinflammatory and profibrotic signaling molecules resulting in renal scarring. The activity of the scarring process with the resultant loss of functional renal mass appears to be modulated, in part, by a polymorphism in the angiotensin converting enzyme (ACE) gene. Clinical studies in adults have demonstrated convincingly the high risk of progression of chronic renal failure (CRF) associated with high BP, the benefit of lowering BP to even the low normal range, and the specific benefit of drugs that inhibit the RAS on the progression of CRF. In children, even moderately elevated BP and moderate proteinuria have been shown to be significant risk factors for progression and CRF. The optimal target BP for children with CRF is currently being determined in a multinational, randomized, prospective trial. PMID- 11866232 TI - Prevalence and consequence of systolic hypertension in children. AB - Systolic blood pressure (SBP) has become the major criterion for the diagnosis, staging, and treatment of hypertension in adults, based on the epidemiology and pathophysiology of adult hypertension, linkage between SBP levels and disease, and benefits of treatment of isolated SBP hypertension. Although children do not typically suffer overt hypertensive disease, an accumulation of data suggests that SBP elevation is as important a factor in the morbidity of hypertension in children as in adults. Systolic BP hypertension is more common in children, whether examining an unselected sampling of patients by routine screening or a selected sampling of referred hypertensive patients. Mild-to-moderate BP elevation in children is associated with increased left ventricular mass (LVM), with SBP more closely linked to LV morphology than diastolic BP (DBP). Furthermore, SBP is associated with increased LVM even in patients with SBP within the normal range. Among hypertensive children, the reported prevalence of left ventricular hypertrophy (LVH) ranges from 30% to 70%, and LVH is more closely related to SBP than to DBP. These data suggest that treatment of hypertension should be directed at normalization of SBP, even when DBP is within the normal range. In addition, trials of antihypertensive medications in children should incorporate SBP hypertension into study inclusion criteria. PMID- 11866233 TI - Cardiovascular sequelae of childhood hypertension. AB - Blood pressure (BP) elevation is a well-established risk factor for cardiovascular morbidity and mortality in adults. More recently, it has been established through autopsy studies and studies using noninvasive imaging techniques that BP elevation is also a risk factor for the development of atherosclerosis in childhood and adolescence. Hypertension is a powerful risk factor for left ventricular hypertrophy and for congestive heart failure in adults. It is now clear that BP elevation is also associated with left ventricular hypertrophy in children and adolescents. If cardiovascular disease morbidity and mortality are to be prevented, then children with elevated BP must be identified and appropriately treated. PMID- 11866234 TI - Blood pressure, hypertension, and ambulatory blood pressure monitoring in children and adolescents. AB - Fixed essential hypertension in children is uncommon. Confusion surrounding the definitions and tools used in measuring blood pressure (BP), and the variable capacity to apply the definitions despite the device used explains this in part. In children, hypertension is defined statistically, based on a large normative population that is ethnically diverse. The standards were developed from the first BP measurement obtained in large studies, and the measurements were obtained using standard auscultatory sphygmomanometry. Technologic advances have seen the widespread introduction of oscillometric devices, which determine BP in a different fashion from auscultation, and the two values are not identical. In fact, as oscillometric devices use proprietary algorithms to calculate the BP, results are not readily interchangeable. Ambulatory BP devices have been added to this mix and offer unique opportunities for accurate diagnosis, and more effective therapies. Most ambulatory BP devices have neither been validated for use in children, nor have passed the validation process. A consensus document may be needed to optimize use and interpretation of data from ambulatory BP monitoring. PMID- 11866235 TI - Arm cuff in the measurement of blood pressure. AB - Selection of an arm cuff of the right size is necessary for accurate measurement of blood pressure (BP). Incorrect measurement can result either in unnecessary investigations, treatment, and follow-up for the mistaken diagnosis of hypertension, or no treatment for hypertension in individuals mistakenly thought to have normal BP. This is of particular importance in children, who have age dependent variations in body size and BP. Studies have revealed that health care providers often do not follow published guidelines concerning cuff selection for children. Perceptions about cuff selection and published recommendations are frequently discrepant. Our studies have revealed that larger BP cuffs are required if upper arm length is used as a criterion for cuff selection. In contrast, smaller cuffs are required if 40% of the upper mid-arm circumference is used as a criterion for cuff selection. No single cuff, when used in accordance with current recommendations, yields accurate systolic as well as diastolic BP. The century-old issue of how to select a right size arm cuff for an accurate measurement of BP remains unresolved. PMID- 11866236 TI - Reproducibility and validity of ambulatory blood pressure monitoring in children. AB - During the past several years ambulatory blood pressure monitoring (ABPM) has been introduced into the study of hypertension and has become a useful tool in making clinical decisions. The ABPM improves the reproducibility of the estimates of a subject's casual blood pressure (BP) both in normotensive and in hypertensive subjects, independent of age. The advantages of ambulatory BP (ABP) over its office counterpart have been studied in children to observe the relationship between BP measurement and early markers of organ damage. In different groups of subjects that have included normotensives, essential hypertensives, renal transplant recipients, or patients with repair of an aortic coarctation, averages of ambulatory systolic BP appear to have the best correlation with left ventricular mass. Moreover, a positive relationship between microalbuminuria and ABP was also observed in a high-risk population with type I diabetes. The assessment of prognostic value of ABPM in children is more difficult because the surrogate end points take time to develop. Pediatric populations with a high risk of developing organ damage markers in a relatively short period of time should be the focus of such studies. Even given its apparent advantage over office BP measurements, much work needs to be carried out so as to better establish the validity and more precise use of ABPM in children. PMID- 11866237 TI - Current concepts in preimplantation genetic diagnosis (PGD): a molecular biologist's view. AB - The first clinically applied preimplantation genetic diagnosis (PGD) was reported more than a decade ago and since then PGD has known an exponential growth. This first report described the use of PCR to sex embryos from couples at risk for X linked diseases. Not surprisingly, in the first years, the development of PCR based tests led to PGD for well-known monogenic diseases such as cystic fibrosis and thalassaemia. When fluorescent in-situ hybridization (FISH) was introduced it quickly replaced PCR-based methods, which had led to misdiagnoses, for sexing of embryos. FISH was also quickly introduced for aneuploidy screening, which has as its main aim the improvement of IVF results in patients with poor reproductive outcome, and later for PGD in translocation carriers. In this review, PGD for patients with a pre-existing genetic risk will be discussed, i.e. the monogenic diseases and the translocations, as well as different biopsy methods and promising new developments. PMID- 11866238 TI - Expression in in-vivo and in-vitro growing and maturing oocytes: focus on regulation of expression at the translational level. AB - Studies of expression in in-vivo and in-vitro maturing oocytes have the potential to elucidate signalling pathways involved in the intricate crosstalk between the oocyte and its somatic compartment during differentiation and morphogenetic processes, and the origin of disturbances in oocyte maturation possibly involved in reduced fertility. This review summarizes data on expression studies with focus on regulation of expression at the translational level in the maturing oocyte. The regulation of gene expression at the translational level as analysed in in-vitro maturing oocytes is complex and highly conserved between different species. It is characterized by differential degradation, and by storage and recruitment of distinct maternal mRNAs involving conserved consensus sequences in the 5' or 3' untranslated regions (UTRs) of mRNAs. Proteins interacting with such sequences affect the temporal 3' polyadenylation, and bring the 5' and 3' UTRs of mRNAs into close proximity for efficient initiation of translation. Post translational modifications of mRNA-associated proteins contribute to maturation- and developmentally controlled and to cell cycle-dependent expression. New methodologies for analysis of ovary-specific gene expression and function of genes in oogenesis are also reviewed, e.g. RT-PCR, SAGE-PCR, real-time rapid cycle fluorescence monitored RT-PCR, differential display techniques, and microinjection of anti-sense RNA, double-stranded RNAs or mRNAs expressing green fluorescent protein-tagged proteins into maturing oocytes. PMID- 11866239 TI - Endometrial factors in recurrent miscarriage. AB - Recurrent pregnancy loss may be a consequence of an abnormal embryonic karyotype, or maternal factors affecting the endometrium resulting in defective implantation. In order to study the endometrial factors responsible for recurrent pregnancy loss, endometrial biopsy samples should be precisely timed according to the LH surge, and the investigation should be carried out in a non-conception cycle, prior to the next pregnancy. The various methods of studying the endometrium including morphological studies, morphometry, immunohistrochemistry, measurement of endometrial protein in plasma and uterine flushings, cytokine expression in endometrial cells, leukocyte populations in the endometrium and ultrasonographic and hysteroscopic studies, were reviewed. The clinical relevance of the observed abnormality depends on whether or not the abnormality is persistent in subsequent cycles, and if the observed abnormality is of significant prognostic value. Very little is known about the treatment of endometrial defect associated with recurrent pregnancy loss, but preliminary data suggest that the use of HMG may be of benefit. PMID- 11866240 TI - Varicocele--the most common cause of male factor infertility? AB - Varicocele is often cited as the most common cause of male factor infertility. Arguments in support of this statement include reports of increased prevalence of varicocele in populations of infertile men compared with fertile or otherwise unselected men, association of varicocele with abnormal semen parameters, and improvements in semen parameters and/or pregnancy rates after varicocele repair. Logically, there would appear to be three possibilities regarding the relationship between varicocele and fertility: (i) varicocele has no association with or effect on male fertility; (ii) varicocele may be associated with, but is not the cause of, male subfertility; and (iii) varicocele is a direct cause of male subfertility. In the following, we review evidence from the literature for and against these three possibilities: at the current time, available evidence appears inadequate to confirm or deny any of these three possibilities. Since the ultimate goal of infertile couples is to conceive, it seem logical that future varicocele research should focus primarily on adequately powered, controlled clinical trials in well-characterized infertile couples, randomized to intervention or appropriate controlled observation, with pregnancy as the primary outcome. PMID- 11866241 TI - Mechanisms of abnormal uterine bleeding. AB - Over the past 10 years there has been an upsurge of interest in the mechanisms underlying normal and disturbed menstrual bleeding. These studies have particularly focused on the mechanisms underlying the common problems of menorrhagia associated with ovulatory and anovulatory dysfunctional uterine bleeding (DUB) and of unpredictable breakthrough bleeding during hormonal contraceptive use. A wide range of abnormalities of endometrial morphology and function have been demonstrated, but it is still not clear how all the pieces of this complex jigsaw puzzle fit together. Ovulatory DUB is predominantly associated with decreased endometrial vasoconstriction and vascular haemostatic plug formation, leading to defective control of the volume of blood which is lost during menstruation. By contrast, breakthrough bleeding is associated with a wide range of molecular disturbances which appear to result in unpredictable vessel breakdown through disturbed endometrial angiogenesis, increased vascular fragility and loss of the integrity of the endothelial, epithelial and stromal supporting structures. Anovulatory DUB is very poorly understood, but may be associated with disturbed angiogenesis, fragile vessels and defective haemostatic processes. Little is known about the actual mechanisms of the common problem of abnormal bleeding associated with specific genital tract pathologies such as uterine myomata. PMID- 11866242 TI - The surgical management of menorrhagia. AB - Surgical treatment of menorrhagia is likely to be both successful and satisfactory to the patient. Correct diagnosis of the cause of menorrhagia is essential, and management should be directed to the specific cause of the problem. The question of which treatment is best is a complex one and involves balancing patient wishes, expected outcomes, complications, cost-effectiveness and quality of life issues. For the subset of women in whom dysfunctional uterine bleeding is diagnosed, the literature suggests that there is a hierarchy of treatments that, in descending order for both efficacy and patient acceptability, are: (i) hysterectomy; (ii) endometrial ablation (either first- or second generation); (iii) the levonorgestrel intrauterine system; and (iv) medical treatments. All four of these options should be discussed with the patient and the relative advantages and disadvantages considered before a treatment decision is made. For patients in whom a pathological cause is diagnosed, specific treatments should be aimed at removal of the lesion and observation of the effect on menstrual status. In addition to the treatment options above, specific treatments such as hysteroscopic, laparoscopic or open excision of the lesion need to be considered. For interventional radiological procedures such as uterine artery embolization, further study is recommended before it can be considered as a safe and effective treatment for menorrhagia. PMID- 11866244 TI - Endometriosis, retrograde menstruation and peritoneal inflammation in women and in baboons. AB - The aim of this review was to assess critically the importance of the frequency, quantity and quality of retrograde menstruation and its relationship to peritoneal inflammation. The basis was the current evidence in women and in baboons supporting the Sampson hypothesis that retrograde menstruation is a key factor in the pathogenesis of endometriosis. It is not proven that retrograde menstruation is a universal phenomenon occurring similarly in women with and without endometriosis. A more thorough understanding of the physiological, cytological and immunological events in peritoneal fluid, peritoneum, endometrium and uterus during menstruation in women with and without endometriosis is critical in order to understand the pathogenesis of this enigmatic disease. PMID- 11866243 TI - Mini symposium on pathogenesis of endometriosis and treatment of endometriosis associated subfertility. Introduction: the endometriosis enigma. PMID- 11866245 TI - The efficacy of medical and surgical treatment of endometriosis-associated infertility: arguments in favour of a medico-surgical aproach. AB - This review discusses the efficacy of a combined, medical (GnRH agonist) and surgical, therapy in endometriosis-associated infertility. Because of the limited information currently available on the activity of lesions in minimal and mild endometriosis, any absolute statement is inappropriate at this time, although some arguments exist in favour of treating endometriosis at laparoscopy. In moderate and severe endometriosis, this review provides arguments in favour of a medico-surgical approach and discusses the possibility of combining medical and surgical therapy. PMID- 11866246 TI - The endometrium versus embryonic quality in endometriosis-related infertility. AB - In spite of a great deal of effort over many decades, the mechanisms that lead to infertility in women with endometriosis remain unknown. Moreover, controversial results in the literature add even more difficulties in the understanding of this issue. Since the introduction of IVF, we have had the opportunity to study the effects of endometriosis at specific stages of the reproductive process including folliculogenesis, fertilization, embryo development and implantation. Depending on the research group, there are conflicting data on the results of IVF in patients with endometriosis. Some researchers found impaired implantation in patients with endometriosis, but such a defect in implantation may be caused by either defective embryos or altered endometrium. The observation of a higher rate of arrested embryos in patients with endometriosis compared with disease-free women, and that women with this pathological situation undergoing ovum donation had the same chance of implantation as patients without endometriosis, initially suggested that impaired oocyte/embryo quality may be responsible for reduced implantation. Further investigations on follicular components have also found differences between women with endometriosis and those without the disease. In addition, recent advances on implantation research, mainly on markers of endometrial receptivity, also show features in the eutopic endometrium of women with endometriosis that are not found in endometrium of women without the disease, although there is no agreement on this point. In this review, we will focus on infertility-related endometriosis based on our own research and the available literature. PMID- 11866247 TI - The brief abstinence test: effects of continued incentive availability on cocaine abstinence. AB - This study compared the effects of 4 voucher incentive conditions of a brief abstinence test on continuous cocaine abstinence. In 3 conditions, cocaine abusing methadone patients could earn $100 for 2 days of cocaine abstinence; 2 of these conditions offered, on either a continuous or interrupted schedule, an additional $300 for evidence of sustained abstinence over the next 9 days. In the 4th condition, no incentives were available. In incentive conditions, 70-80% of patients initiated abstinence, compared with 48% in the no-incentive condition. Both continuing reinforcement conditions produced higher rates of sustained abstinence than the single and no-voucher conditions. The study confirmed the utility of quantitative urine-testing methods combined with high valued incentives to promote cocaine abstinence initiation in methadone maintenance patients. PMID- 11866248 TI - Anxiolytic effects of mecamylamine in two animal models of anxiety. AB - Clinical and preclinical evidence suggests that mecamylamine, a nicotinic receptor antagonist, may have anxiolytic properties. The purpose of this study was to further investigate the anxiolytic properties of mecamylamine in rats as measured by the Elevated Plus Maze and the Social Interaction models of anxiety and to determine if manipulation of the testing environment (either brightly lit or dimly lit conditions) influenced the results. Results indicated that mecamylamine had significant anxiolytic effects in both the Elevated Plus Maze and Social Interaction Tests and that these effects were dependent on dose administered and the level of anxiety produced under different testing conditions. If confirmed by further clinical research, nicotinic receptor antagonists like mecamylamine may represent a novel class of anxiolytics. PMID- 11866250 TI - The observing-response procedure: a novel method to study drug-associated conditioned reinforcement. AB - In this experiment, the observing-response procedure was adapted for use with drug self-administration. Rats' responding for oral ethanol was sometimes reinforced on a random-ratio schedule, whereas at other times it had no effect (i.e., extinction). Behavior producing stimuli associated with the otherwise unsignaled random-ratio and extinction periods (i.e., observing behavior) was acquired and maintained. In a vehicle control condition, both self-administration and observing behavior decreased, but observing decreased less rapidly proportionally to baseline than vehicle consumption. Thus, conditioned reinforcers may have persistent effects that are relatively independent of the current status of the primary reinforcer. The procedure allows long-term study of drug-associated conditioned reinforcement and provides independent indexes of the conditioned reinforcing and discriminative stimulus effects of drug stimuli. PMID- 11866249 TI - Possible amotivational effects following marijuana smoking under laboratory conditions. AB - Human participants earned money by responding on a progressive-ratio (PR) schedule (initial value $50) or received money without responding on a fixed-time (FT) schedule. During the session, participants could terminate the PR schedule and initiate an FT 200-s schedule. In Experiment 1, increases in monetary value produced increased number of responses, time spent, and money earned in the PR component. In Experiment 2, marijuana smoking produced potency-related reductions in the number of responses, time spent, and money earned in the PR component, effects that can be interpreted as amotivational. Increasing the monetary value of the reinforcer diminished the acute marijuana effects on PR responding, suggesting that marijuana exerted an effect primarily on reinforcers of a smaller magnitude. PMID- 11866251 TI - Heroin craving and drug use in opioid-maintained volunteers: effects of methadone dose variations. AB - Recent data indicate that opioid agonist and antagonist challenges decrease and increase (respectively) heroin craving in physically dependent individuals. This study investigated effects of methadone dose variations on craving and new drug use in 18 outpatients who were given money contingencies. In Phase 1, volunteers were told in different test sessions that methadone dose would increase, decrease, or stay the same; drug-abstinence contingencies were suspended for 24 hr. Craving significantly increased and new heroin use marginally increased (relative to maintenance dose) only when a dose reduction was paired with a dose decrease instruction. In Phase 2 (detoxification), craving and heroin use significantly increased as methadone dose decreased. Thus, loss of micro-receptor agonist effect increased craving and risk of relapse. PMID- 11866252 TI - Contingency management to enhance naltrexone treatment of opioid dependence: a randomized clinical trial of reinforcement magnitude. AB - Fifty-five detoxified opioid-dependent individuals were randomly assigned to 1 of 3 treatments delivered over 12 weeks: standard naltrexone maintenance, standard naltrexone plus low-value contingency management (CM), or standard naltrexone plus high-value CM. Results suggest that (a) assignment to either CM condition was associated with significant reductions in opioid use over time compared with standard naltrexone treatment; (b) contrasts of high- versus low-value reinforcement magnitude were not significant, suggesting no relative benefit of higher over lower value incentives in this population; (c) participants assigned to either CM group reported significant reductions in readiness to change compared with participants assigned to standard naltrexone treatment. These findings suggest that targeted behavioral therapies can play a substantial role in broadening the utility of available pharmacotherapies. PMID- 11866253 TI - Rectal bleeding: is it always an indication for colonoscopy? PMID- 11866254 TI - Screening for hepatocellular carcinoma--answers to some simple questions. PMID- 11866255 TI - A population-based cohort study comparing laparoscopic and open cholecystectomies: research in the new millennium. PMID- 11866256 TI - Anorectal functional testing: review of collective experience. AB - Anorectal manometry includes a number of specific tests that are helpful in the diagnostic assessment of patients with fecal incontinence and constipation; their purpose is to delineate the pathophysiological mechanism for these symptoms. Some of these tests may also provide helpful information in the assessment of patients with rectal pain or diarrhea, but their sensitivity and specificity are less well established for these symptoms. Tests for which there is consensus regarding their clinical utility include 1) resting anal canal pressure, 2) anal canal squeeze pressure (peak pressure and duration), 3) the rectoanal inhibitory reflex elicited by balloon distension of the rectum, 4) anal canal pressure in response to a cough, 5) anal canal pressure in response to defecatory maneuvers, 6) simulated defecation by means of balloon or radiopaque contrast, 7) compliance of the rectum in response to balloon distension, and 8) sensory thresholds in response to balloon distension. Anal endosonography and pelvic floor electromyography from intra-anal plate electrodes are nonmanometric tests that are also specifically useful in the evaluation of constipation and fecal incontinence. The clinical utility of all anorectal manometric tests is limited by the relative absence of 1) standardization of test protocols and 2) normative data from a large number of healthy individuals. The interpretation of these diagnostic tests is also complicated by the fact that patients are able to compensate for deficits in specific physiological mechanisms maintaining continence and defecation by utilizing other biological and behavioral mechanisms. PMID- 11866257 TI - Brazilian consensus on gastroesophageal reflux disease: proposals for assessment, classification, and management. AB - The Brazilian Consensus on Gastroesophageal Reflux Disease considers gastroesophageal reflux disease to be a chronic disorder related to the retrograde flow of gastroduodenal contents into the esophagus and/or adjacent organs, resulting in a variable spectrum of symptoms, with or without tissue damage. Considering the limitations of classifications currently in use, a new classification is proposed that combines three criteria-clinical, endoscopic, and pH-metric-providing a comprehensive and more complete characterization of the disease. The diagnosis begins with the presence of heartburn, acid regurgitation, and alarm manifestations (dysphagia, odynophagia, weight loss, GI bleeding, nausea and/or vomiting, and family history of cancer). Also, atypical esophageal, pulmonary, otorhinolaryngological, and oral symptoms may occur. Endoscopy is the first approach, particularly in patients over 40 yr of age and in those with alarm symptoms. Other exams are considered in particular cases, such as contrast radiological examination, scyntigraphy, manometry, and prolonged pH measurement. The clinical treatment encompasses behavioral modifications in lifestyle and pharmacological measures. Proton pump inhibitors in manufacturers' recommended doses are indicated, with doubling of the dose in more severe cases of esophagitis. The minimum time of administration is 6 wk. Patients who do not respond to medical treatment, including those with atypical manifestations, should be considered for surgical treatment. Of the complications of gastroesophageal reflux disease, Barrett's esophagus presents a potential development of adenocarcinoma; biopsies should be performed, independent of Barrett's esophagus extent or location. In this regard the designation "short Barrett's" is not important in terms of management and prognosis. PMID- 11866258 TI - Complications of gallstone disease: Mirizzi syndrome, cholecystocholedochal fistula, and gallstone ileus. AB - Gallstone is a common disease with a 10% prevalence in the United States and Western Europe. However, it is only symptomatic in 20-30% of patients, with biliary pain "colic" being the most common symptom. Complications of asymptomatic gallstone disease are generally rare, with an incidence of <1 %/yr. The most common complications of gallstone disease are acute cholecystitis, acute pancreatitis, ascending cholangitis, and gangrenous gallbladder. Less frequent complications include Mirizzi syndrome, cholecystocholedochal fistula, and gallstone ileus. Mirizzi syndrome and cholecystocholedochal fistula are two manifestations of the same process that starts with impaction of a gallstone in the gallbladder neck that results in obstruction of the bile duct, causing jaundice. The gallstone may erode into the bile duct, causing cholecystocholedochal fistula. Gallstone ileus refers to small bowel obstruction resulting from the impaction of one or more gallstones after they have migrated through a cholecystoenteric fistula. An accurate diagnosis is essential to the management and prevention of further complications. A variety of imaging and endoscopic modalities are used to make the diagnosis once the condition is suspected clinically. Treatment should be tailored to each individual patient. Management choices include ERCP, lithotripsy (endoscopic or extracorporeal), and surgery. Prognosis is frequently related to early recognition, management of any comorbid conditions, and careful selection of treatment modalities. PMID- 11866259 TI - Anorexia nervosa: manifestations and management for the gastroenterologist. AB - Anorexia nervosa is a complex psychiatric disorder with significant morbidity and mortality. It is important for gastroenterologists to be aware of the physiological effects and potential complications of anorexia nervosa, as they are frequently involved in treating patients with this disorder. We review the classic, GI, and neuroendocrinological features of anorexia nervosa. We also discuss gender differences and treatment options in anorexia nervosa. Further studies of GI physiology and pharmacology are needed to determine whether any disturbances may be amenable to therapeutic intervention. Future treatments directed at improving GI sensorimotor function and neurohormonal abnormalities in patients with anorexia nervosa may impact their nutritional rehabilitation and may have important health economic implications as patients avoid hospitalization and are restored to full activities in society. The current team approach, which incorporates psychiatrists, psychologists, nutritionists, pediatricians, internists, and gastroenterologists in the treatment of patients with anorexia nervosa, will continue to be essential. PMID- 11866260 TI - Impact of esophageal acid exposure on the endoscopic reversal of Barrett's esophagus. AB - OBJECTIVE: Normalization of esophageal acid exposure had been assumed to be necessary for the reversal of Barrett's esophagus with endoscopic therapy. This assumption is examined by evaluating the esophageal pH in a group of patients undergoing reversal therapy with fixed high-dose proton pump inhibitor therapy and endoscopic multipolar electrocoagulation (MPEC). METHODS: Patients with Barrett's esophagus of 2-6 cm in length were treated with omeprazole (40 mg b.i.d.). They underwent 24-h esophageal pH monitoring 5 cm above the upper margin of the lower esophageal sphincter determined by a water-perfused catheter. They then underwent MPEC therapy to an endpoint of elimination of Barrett's (reversal) by both endoscopy and biopsy 6 months after the last MPEC session or failure to achieve visual (endoscopic) reversal after six treatment sessions. RESULTS: Twenty patients had 24-h pH testing and reached a reversal endpoint. Three patients had abnormal pH tests, two total and supine and one supine only. These patients had documented reversal. The remaining 17 patients had normal pH testing but five failed reversal therapy. CONCLUSION: Barrett's esophagus can be completely reversed with endoscopic therapy despite abnormal esophageal acid exposure. Also, patients can fail reversal even with normal esophageal acid exposure. The necessary reduction of esophageal acid exposure for reversal therapy has yet to be defined. PMID- 11866261 TI - Barrett's esophagus and reflux esophagitis: is there a missing link? AB - OBJECTIVES: Barrett's esophagus (BE) is associated with esophageal reflux. The development stage of BE is not well described. Epidemiological evidence indicates that the columnar epithelium in BE is acquired and reaches its full length rapidly. We tested the hypothesis that BE might result from direct replacement of erosions in reflux esophagitis (RE). METHODS: At endoscopy, we compared the length and distribution of esophageal erosions in 50 patients with RE with the length and distribution of columnar epithelium in 50 patients with BE. RESULTS: The median length of erosions in RE was 2 cm, less than the median length of columnar epithelium in BE, 5 cm (p < 0.001). Erosions in RE were usually multiple and scattered, involving the entire circumference of the esophagus in only 10% of cases, but circumferential involvement by columnar epithelium was found in 68% of BE cases (p < 0.001). Circumferential involvement, 3 cm or longer, was found in 0% of cases of RE versus 56% of BE cases (p < 0.001). Two patients without RE or BE had large areas of epithelial loss of uncertain etiology. CONCLUSIONS: The length and distribution of erosions in RE differ greatly from the length and distribution of columnar epithelium in BE. It is unlikely that BE arises directly from areas of esophagitis. We suggest that BE may develop after loss of a long segment of squamous epithelium, with columnar replacement in the presence of continuing acid reflux. PMID- 11866262 TI - The adoption of ablation therapy for Barrett's esophagus: a cohort study of gastroenterologists. AB - OBJECTIVES: Although ablation therapy may be useful in the treatment of Barrett's esophagus. evidence of effectiveness is scarce, and little is known about current utilization of ablation. We aimed to determine whether the use of ablation was increasing in a cohort of gastroenterologists, and to identify physician beliefs and characteristics associated with ablation use. METHODS: We surveyed a national sample of gastroenterologists about ablation use, with an 18-month follow-up. The self-administered instrument included questions about demographic characteristics, attitudes about ablation therapy, and prior experience with ablation. Case scenarios were also included. We used logistic regression to identify factors associated with the use of ablation in patients with Barrett's esophagus. RESULTS: Two hundred seventy-nine (50.3% of those eligible) responded to the baseline survey. Few agreed that ablation lowers the risk of adenocarcinoma (15%) or is supported by the medical literature (19%). However, 25% of respondents reportedly had used ablation at baseline, and this increased to 36% in the follow-up survey (p = 0.0003). The use of ablation was significantly associated with physician age greater than 54 yr (odds ratio [OR] = 2.77, 95% CI = 1.04-7.37) and the belief that ablation was used by colleagues (OR = 13.27, 95% CI = 4.44-39.64) or decreases medical costs (OR = 5.07, 95% CI = 1.00-25.74). CONCLUSIONS: Although few gastroenterologists agreed that ablation is effective, a significant proportion had adopted its use. There was a significant increase in ablation use during our study period, and the characteristic that was most strongly associated with ablation use was the belief that colleagues used it. PMID- 11866263 TI - Esophageal tuberculosis: is it so rare? Report of 12 cases and review of the literature. AB - OBJECTIVE: Mycobacterial involvement of the esophagus is rare in both immunocompetent and immunocompromised hosts with advanced pulmonary tuberculosis, even in countries with high prevalence rates like India. Most of the reported cases of esophageal tuberculosis (ET) are secondary to tuberculosis elsewhere in the body, most commonly pulmonary tuberculosis. Very few cases of isolated or primary ET have been reported, and most of them have been from developing countries. The upsurge in reported cases of tuberculosis linked to the AIDS epidemic has increased the incidence of this infection in developed countries also. Our aim is to study tuberculosis as an etiological factor in the causation of dysphagia, the role of cytology and histopathology in establishing its diagnosis, and the outcome of antitubercular treatment in these patients. METHODS: A hospital-based, retrospective study was performed. We reviewed records of all of the patients who underwent upper GI endoscopic examination for complaints of persistent dysphagia (>6 wk) in a tertiary care hospital in India between 1995 and 1999. Patients with abnormal endoscopic findings were subjected to endoscopic fine needle aspiration/brush cytology and biopsies. Those with pathological findings suggestive of tuberculosis were treated with antitubercular therapy. RESULTS: Tubercular involvement of the esophagus, confirmed by pathological examination, was found in 12 patients. They constituted 0.5% of all patients with persistent dysphagia, and 1.3% of all patients having abnormal esophagoscopic findings. Cytological examination provided a very useful diagnostic parameter in detection of these cases. Patients diagnosed as having ET responded well to antitubercular therapy. CONCLUSIONS: Tuberculosis as a causative factor for dysphagia should be considered in developing countries with high incidences of tuberculosis and in immunocompromised hosts. Detection of these cases by careful examination of cytological smears and biopsies and treatment with standard antitubercular therapy appear effective. PMID- 11866264 TI - Conservative treatment of primary gastric low-grade B-cell lymphoma of mucosa associated lymphoid tissue: predictive factors of response and outcome. AB - OBJECTIVES: Primary gastric low-grade B-cell lymphoma of mucosa-associated lymphoid tissue may regress with conservative treatment such as anti Helicobacterpylori therapy or monochemotherapy. The aims of the present study were to analyze the predictive factors of response to anti-H. pylori treatment, to assess the effects of an adjuvant therapy in responding patients, and to evaluate an alternative therapy in nonresponding patients. METHODS: From 1995 to 2000, 48 H. pylori-infected patients with localized primary gastric low-grade B cell lymphoma of mucosa-associated lymphoid tissue were treated with anti-H. pylori therapy. Endoscopic and endoscopic ultrasonography features and histological grading of large cells' proportion were analyzed. Eradication of H. pylori and tumoral response were assessed at 2 and 6 months, respectively. From 1996, patients in remission at 6 months were randomized to receive either chlorambucil p.o. for 6 months or no treatment. Patients who did not respond to H. pylori eradication received chlorambucil p.o. for 1 yr. RESULTS: Among the 48 treated patients, 33 (69%) were in complete (n = 28) or in partial (n = 5) remission, and 15 (31%) were in treatment failure at 6 months. H. pylori was eradicated in 47 patients. The response was not correlated with the endoscopic features or with the histological grade. In contrast, it was related to ultrasonographic features: remission was achieved in 76% of patients when no perigastric lymph node was detected versus only 33% when endoscopic ultrasonography showed presence of lymph nodes (p = 0.025). All responding patients remained in remission (median 34 months) whatever the treatment they received (no treatment or chlorambucil). Remission could be achieved with chlorambucil in 58% of the nonresponding patients to anti-H. pylori treatment. CONCLUSIONS: The major negative predictive factor of the tumoral response to anti H. pylori treatment in patients with primary gastric low-grade B-cell lymphoma of mucosaassociated lymphoid tissue was the presence of perigastric lymph nodes on endoscopic ultrasonography. In responding patients, remission remained stable, suggesting that adjuvant chemotherapy was not useful. In patients who failed to respond to H. pylori eradication, monochemotherapy with chlorambucil proved to be efficient, but new therapeutic modalities should be evaluated to improve the control of the tumoral process. PMID- 11866265 TI - Gastric polyps in pediatrics: an 18-year hospital-based analysis. AB - OBJECTIVE: Gastric polyps are recognized as either an incidental finding on routine gastroscopy or a frequent occurrence in patients with polyposis syndromes. The aim of this study is to determine the prevalence, clinical presentation, and histological subclassification of gastric polyps identified on esophagogastroduodenoscopy in pediatric patients. METHODS: We performed an 18-yr retrospective study of all pediatric (<21 yr) patients with gastric polyps diagnosed between 1983 and 2000 at The Johns Hopkins Children's Center. The histology slides were all evaluated at the time of the study according to the accepted histological classification of gastric polyps. RESULTS: Gastric polyps were reported in 40 procedures (0.7%) [corrected] performed in 35 (male:female 1.3:1) patients with a mean (SEM) age at diagnosis of 14.4 (0.9) yr. Polyps were more frequent in white than in black patients (adjusted ratio 1.4:1). The histological subtypes included hyperplastic-inflammatory (42%), fundic gland (40%), hamartomatous (10%), adenomatous (5%), and heterotopic polyps (3%). Fundic gland polyps were frequently encountered in patients with familial adenomatous polyposis (81%). These patients tended to be asymptomatic at the time of their surveillance esophagogastroduodenoscopy, and frequently harbored histological changes of either dysplasia (31%) or indeterminate of dysplasia (19%). CONCLUSIONS: Hyperplastic polyps are the most frequently identified gastric polyps in our pediatric population. Fundic gland polyps are common in patients with familial adenomatous polyposis wherein they tend to harbor histological changes of dysplasia. Future longitudinal studies are needed to evaluate the temporal progression of dysplasia to gastric cancer in patients with fundic gland polyps, and to establish esophagogastroduodenoscopy surveillance guidelines. PMID- 11866266 TI - Helicobacter pylori infection, not gastroesophageal reflux, is the major cause of inflammation and intestinal metaplasia of gastric cardiac mucosa. AB - OBJECTIVE: The etiology of inflammation below the normal Z-line is an area of intense debate. Some suggest this is the earliest change of chronic gastroesophageal reflux disease (GERD), whereas others indict Helicobacter pylori (H. pylori) as the main cause. The aim of this study was to evaluate the relationship among inflammation of gastric cardiac mucosa (carditis), H. pylori infection, and intestinal metaplasia in patients with GERD and Barrett's esophagus compared with age-matched controls. METHODS: Patients with GERD and Barrett's esophagus were compared with controls undergoing endoscopy for a variety of other conditions. Endoscopic biopsy specimens from the gastric cardia (obtained on retroflexed view), fundus, and antrum were evaluated for inflammation, H. pylori infection, and intestinal metaplasia. RESULTS: The prevalence of H. pylori infection did not significantly differ among the study populations: controls (42%), GERD (33%), and Barrett's esophagus (27%) (p = 0.20). However, the prevalence of carditis significantly decreased from the control group (30%) to those with GERD (23%) and Barrett's esophagus (11%) (p = 0.03). Overall, 42 of 51 (82%) patients with carditis had H. pylori; all had pangastritis. The prevalence of cardia intestinal metaplasia also significantly decreased from the control group (15%) to those with GERD (4%) and Barrett's esophagus (0%) (p = 0.003). Of 13 patients with cardia intestinal metaplasia, 12 had carditis, 10 had H. pylori infection, and seven had intestinal metaplasia elsewhere in the stomach. CONCLUSIONS: Inflammation of gastric cardiac mucosa decreases in prevalence from controls to patients with GERD and Barrett's esophagus and correlates strongly with H. pylori infection. Cardia intestinal metaplasia is associated with H. pylori-related cardiac inflammation and intestinal metaplasia elsewhere in the stomach. PMID- 11866267 TI - Cytokine gene profile in gastric mucosa in Helicobacter pylori infection and Zollinger-Ellison syndrome. AB - OBJECTIVE: Helicobacter pylori-associated inflammation is mediated, in part, by inflammatory cytokines. In contrast, the mucosal disease associated with Zollinger-Ellison syndrome (ZES) is acid driven, and the role of cytokines is not known. The aim of this study was to elucidate the role of cytokines in these two diseases, as we quantitated proinflammatory cytokine messenger RNA (mRNA) levels in the gastric mucosa from patients with H. pylori infection and ZES. METHODS: The study population included 11 patients with H. pylori infection, 12 with ZES, 17 with both H. pylori infection and ZES, and three control subjects with neither. Using a competitive polymerase chain reaction for interleukin (IL) 1beta, IL-6, IL-8, and tumor necrosis factor-alpha, the polymerase chain reaction products in gastric biopsies were quantitated by capillary electrophoresis and laser-induced fluorescence. RESULTS: The levels of IL-1beta, IL-6, IL-8, and tumor necrosis factor-a mRNA in gastric tissue of patients with H. pylori infection only and ZES only exceeded the levels in the control gastric tissue (p < 0.05 to p < 0.005). Unexpectedly, the number of molecules of IL-1beta and IL-8 mRNA in gastric tissue from ZES patients exceeded the levels in gastric tissue from patients with H. pylori only (p < 0.05). The local levels of cytokine mRNA in patients with both diseases exceeded the levels in patients with H. pylori only (IL-6, p < 0.05; IL-8, p < 0.05) and ZES only (IL-6, p < 0.05; tumor necrosis factor-a, p < 0.05). CONCLUSIONS: Levels of proinflammatory cytokine mRNA are increased in acid-driven as well as infection-driven gastric inflammation, and the presence of both disease processes appears to have an additive effect on local cytokine message expression. Inflammatory cytokines may mediate both infection- and acid-driven gastric inflammation. PMID- 11866268 TI - Cortical mapping of visceral pain in patients with GI disorders using functional magnetic resonance imaging. AB - OBJECTIVE: We sought to identify central loci that activate in response to visceral stimuli (stool and pain). We had a particular interest in observing the anterior cingulate gyrus and frontal cortex in normals and in patients with intestinal disease, including inflammatory bowel disease (IBD) and irritable bowel syndrome (IBS). METHODS: Subjects underwent rectal balloon distention to a sensation of stool and to a sensation of pain while undergoing blood oxygenation level-dependent functional magnetic resonance imaging. Experiments were conducted in a Magnex 3.0-T whole body magnet with a Bruker Biospec console and a quadrature head coil. Four contiguous 5.0-mm oblique axial slices designed to optimize coverage of areas believed to be responsive to noxious stimulation were acquired. Activations were detected by using cross-correlation maps (p < 0.001) for individual subjects. The experimental groups were compared using both an analysis of variance and profile analysis. RESULTS: A significantly higher percentage of pixels activated in the anterior cingulate gyrus over both pain and stool conditions for the control group than for the IBS group and for the IBS group than for the IBD group (p < 0.035). Deactivation of left somatosensory cortex was greater for the IBS group than for the IBD group and greater for the IBD group than for the controls (p < 0.0065) in the boxcar condition. Frontal deactivation in controls compared with disease groups bordered on statistical significance. Profile analysis of the three groups across six regions of interest revealed that the control and IBD groups were distinguished by different profiles of response (p < 0.005). Nonparametric evaluation of the data suggests that, among the pixels in the anterior cingulate activating to pain, there are two patterns of response to pain-on/off and graded. This was true for both controls and disease groups. CONCLUSIONS: Normal controls and subjects with IBD and IBS share similar loci of activations to visceral sensations of stool and pain. Both activation and deactivation of particular regions of interest differentiate the three groups, as do profiles of patterned response across six of the regions of interest for the control and IBD groups. PMID- 11866269 TI - Yield of colonoscopy in patients with nonacute rectal bleeding: a multicenter database study of 1766 patients. AB - OBJECTIVE: There are few data to guide the choice between colonoscopy and flexible sigmoidoscopy in patients with nonacute rectal bleeding, especially in younger age groups. Our aim was to determine the yield of colonoscopy for significant proximal large bowel disease in the absence of significant distal disease, with special reference to young patients. METHODS: This was a retrospective study of data collected prospectively in 1766 patients (median age 57 yr, 711 women). The endoscopic database (GI-Trac) contained 152 discrete fields for data input. Multiple logistic regression analysis was performed to identify variables independently associated with the presence of isolated significant proximal disease. RESULTS: Young patients had a higher percentage of normal examinations than did older patients. The incidence of diverticular disease, small polyps, large polyps, and cancer rose with increasing age. No patient aged <40 yr had an isolated proximal cancer, but 7% had other significant isolated proximal disease. There was no overall association between age and significant proximal disease in the absence of significant distal disease (p = 0.66). The only variable associated with isolated proximal disease was anemia (odds ratio = 1.81; 95% CI = 1.11-2.93; p = 0.02). CONCLUSION: The yield of colonoscopy (beyond the range of sigmoidoscopy) for neoplasia is low in patients aged <40 yr, but other significant disease may be missed if age is the only criterion determining colonoscopy use. PMID- 11866270 TI - A population-based cohort study comparing laparoscopic cholecystectomy and open cholecystectomy. AB - OBJECTIVES: Laparoscopic cholecystectomy (LC) has become a popular alternative to open cholecystectomy (OC). Previous studies comparing outcomes in LC and OC used small selected cohorts of patients and did not control for comorbid conditions that might affect outcome. The aims of this study were to characterize the morbidity, mortality, and costs of LC and OC in a large unselected cohort of patients. METHODS: We used the population-based North Carolina Discharge Abstract Database (NCHDAD) for January 1, 1991, to September 30, 1994 (n = 850,000) to identify patients undergoing OC and LC. We identified the indications for surgery, complications, and type of perioperative biliary imaging used. We compared length of stay, hospital charges, complications, morbidity, and mortality between OC and LC patients. To account for variations in outcomes from differences in age and comorbidity between the OC and LC groups, we used the age adjusted Charlson Comorbidity Index in regression analyses quantifying the association between type of surgery and outcome. RESULTS: Our cohort consisted of 43,433 patients (19,662 LC and 23,771 OC). The mean age-adjusted Charlson Comorbidity Index score was slightly higher for the OC compared to the LC group (4.3 vs 4.1, p < 0.05). The OC patients had longer hospitalizations, generated more charges ($12,125 vs $9,139, p < 0.05), and required home care more often. The crude risk ratio comparing risk of death in OC to LC was 5.0 (95% CI = 3.9 6.5). After controlling for age, comorbidity, and sex, the odds of dying in the OC group was still 3.3 times (95% CI = 1.4-7.3) greater than in the LC group. In the LC group, the number of patients with acute cholecystitis rose over the study period, whereas the number of patients with chronic cholecystitis declined. In the OC group, the number of patients with acute and chronic cholecystitis declined. The use of intraoperative cholangiography was greater in the OC group but declined in both groups over the study period. The use of ERCP was greater in the LC group and increased in both groups over time. CONCLUSIONS: The introduction of LC has resulted in a change in the management of cholecystitis. Despite a higher proportion of patients with acute cholecystitis, the risk of dying was significantly less in LC than in OC patients, even after controlling for age and comorbidity. Based on lower costs and better outcomes, LC seems to be the treatment of choice for acute and chronic cholecystitis. PMID- 11866271 TI - Mutational screening of patients with nonalcoholic chronic pancreatitis: identification of further trypsinogen variants. AB - OBJECTIVES: Mutations of the cationic trypsinogen (CT) and the serine protease inhibitor, Kazal type 1 (SPINK 1) are associated with chronic pancreatitis. After mutational screening of a cohort of patients with nonalcoholic chronic pancreatitis, we report three novel variants of the trypsinogen molecule and the clinical characteristics of the carriers. METHODS: The coding region of the exon 2 and 3 of the CT gene of 523 patients with chronic nonalcoholic pancreatitis (108 patients with suspected hereditary pancreatitis (HP) and 415 patients with "idio pathic" pancreatitis [IP]) and 82 controls was analyzed after polymerase chain reaction amplification. Clinical characteristics were obtained by questioning the patients and their relatives and physicians. HP was suspected when two members of a family had chronic pancreatitis. A restriction digestion was used to analyze the N34S mutation SPINK1. RESULTS: The mutation R122H of the cationic trypsinogen was found in 21 index patients, N291 in six index patients, and A16V and D22G in one index patient, all from HP families. The N34S mutation of SPINK1 was found in two index patients with a family history of HP. In three patients, the novel point mutations L104P, R116C, and C139F of the cationic trypsinogen were found. A clear autosomally dominant inheritance of chronic pancreatitis was not present in these families. In 75 index patients from HP families (69.4%), no mutation could be found. The SPINK 1-mutation N34S was detected in only one patient carrying a CT mutation, and was found in 68 (16.4%) of patients with IP. CONCLUSIONS: The R122H and N291 mutations of CT are the most common disease-associated mutations in HP; the N34S mutation of SPINK I is the most frequent genetic risk factor associated with IP. The CT gene carries several variations that could be associated with chronic pancreatitis. To avoid overestimating the pathogenetic impact of novel trypsinogen variants, a detailed clinical characterization of all patients with early onset chronic pancreatitis is mandatory. PMID- 11866272 TI - Comparison between magnetic resonance cholangiopancreatography and ERCP for evaluation of the pancreatic duct. AB - OBJECTIVE: To date, ERCP has been the reference technique in the diagnosis of pancreatic duct pathology. The aim of the present study was to evaluate the accuracy of magnetic resonance cholangiopancreatography (MRCP), a relatively new, noninvasive diagnostic method, in studying the pancreatic duct. METHODS: A prospective 15-month study was done of 78 patients with suspected biliopancreatic pathology and indications for ERCP. In all cases, MRCP was performed <72 h before ERCP. Both techniques were used to assess the size of the pancreatic tract (normal or dilated), the presence or absence of obstruction, the level of obstruction where present, and its etiology. RESULTS: Both techniques found the pancreatic tract to be normal and nondilated in 60 patients. The specificity and sensitivity of MRCP in evaluating the normal pancreatic duct were 98% and 94%, respectively. In nine cases, a stenotic duct of Wirsung was detected (seven at the head of the pancreas and two in the body), with the same correlation being shown by ERCP (sensitivity 100%). The sensitivity and specificity of MRCP relating to diffuse irregular and dilated morphologies of the entire pancreatic duct was 86% and 100%, respectively. Nine patients were diagnosed with pancreatic neoplasms by both techniques, with an accuracy of 100%. Four patients presented chronic pancreatitis, which was correctly diagnosed by MRCP in each case. CONCLUSIONS: MRCP is an effective technique for evaluating both the normal and diseased duct of Wirsung, and for establishing the underlying pathology. PMID- 11866273 TI - Plain abdominal radiographic features are not reliable markers of disease extent in active ulcerative colitis. AB - OBJECTIVES: The aims of this study were to assess the reliability of the distribution of fecal residue, mucosal irregularity, and colonic wall thickening on plain abdominal x-ray as markers of disease extent assessed by technetium-99 m hexamethylpropylene amine oxine-labeled leukocyte scans in active ulcerative colitis (UC). METHODS: Plain abdominal radiographs were blindly assessed for the distribution of fecal residue, mucosal irregularity, and colonic wall thickening (>3 mm) in 30 patients with active UC. Most patients were too sick to safely allow total colonoscopy. In 11 patients for whom details of disease extent were available from total colonoscopy or surgery undertaken at the time, there was a close correlation with disease extent defined by technetium-99 m hexamethylpropylene amine oxine-labeled leukocyte scans. Contemporaneous radiolabeled leukocyte scans were therefore used to assess disease extent in comparison with plain abdominal radiographs. RESULTS: Of 30 patients, 15 had pancolitis and 15 had subtotal or distal disease as indicated by radiolabeled leukocyte scans. The distribution of fecal residue on plain abdominal radiographs correctly identified disease extent defined on radiolabeled leukocyte scans in 40% of patients, overestimating it in 13% and underestimating it in 47%. There was no significant correlation between distribution of fecal residue on plain abdominal radiographs and disease extent on radiolabeled leukocyte scans or colonoscopy or surgery. Of patients with pancolitis, 60% had fecal residue present on plain abdominal radiograph with 40% showing stool distal to the hepatic flexure as well as in the right colon. For total UC on radiolabeled leukocyte scanning, the sensitivity and specificity of absence of fecal residue on plain abdominal radiographs were 40% and 80% respectively. Irregularity of mucosal edge and colonic wall thickening were even less accurate than fecal residue in defining disease extent. CONCLUSIONS: The distribution of fecal residue, irregularity of mucosal edge, and colonic wall thickening on plain abdominal radiography do not provide a reliable guide to disease extent in active UC. PMID- 11866274 TI - Lactoferrin in whole gut lavage fluid as a marker for disease activity in inflammatory bowel disease: comparison with other neutrophil-derived proteins. AB - OBJECTIVES: We investigated which neutrophil-derived proteins in whole gut lavage fluid (WGLF) most accurately reflect disease activity in inflammatory bowel disease. METHODS: WGLF was obtained from patients undergoing whole gut lavage as a bowel preparation for colonoscopy. Twenty-seven patients with ulcerative colitis (UC), 23 patients with Crohn's disease (CD), and 35 control subjects were examined. The concentrations of lactoferrin, polymorphonuclear neutrophil elastase (PMN-E), myeloperoxidase, and lysozyme in WGLF were measured by ELISA. For the assessment of stability, WGLF samples were stored at 37 degrees C for various periods. RESULTS: In UC, the concentrations of lactoferrin, myeloperoxidase, and lysozyme in WGLF had good correlations with colonoscopic grading. Zero, 12, five, and 10 of 28 samples from active UC patients showed normal concentrations of lactoferrin, PMN-E, myeloperoxidase, and lysozyme, respectively. In CD, the concentrations of lactoferrin and myeloperoxidase had good correlations with the Crohn's disease activity index. Thirteen and seven of 36 samples from inactive CD patients (Crohn's disease activity index < or = 150) showed high concentrations of lactoferrin and myeloperoxidase, respectively. Most of them (11/13, 6/7) were found to have ulceration by colonoscopy or small bowel x-ray. The ratio of the lactoferrin concentration in the WGLF supernatant to that in total WGLF was highest among these proteins in all disease groups and control subjects. Lactoferrin and myeloperoxidase showed good stability in WGLF, whereas PMN-E and lysozyme did not. CONCLUSION: Lactoferrin is the most suitable of these proteins for use as a neutrophil-derived WGLF marker of intestinal inflammation. PMID- 11866275 TI - Prolapsing mucosal polyps: an underrecognized form of colonic polyp--a clinicopathological study of 15 cases. AB - OBJECTIVE: Prolapsing intestinal mucosa occurs in many forms throughout the GI tract. We describe 15 patients with polypoid masses in the sigmoid colon and histological features of mucosal prolapse. METHODS: Fifteen patients with colon polyps demonstrating endoscopic and histological features of mucosal prolapse were retrospectively identified from our database. RESULTS: Twelve patients presented with signs and symptoms that were nonspecific, but consistent with mucosal prolapse, such as occult or gross intestinal bleeding and lower abdominal pain. Three patients were asymptomatic. The polyps occurred in the sigmoid colon, usually in association with diverticular disease, and appeared more often in men. Endoscopically, the polyps appeared to be well-circumscribed, hyperemic masses that contrasted sharply with normal-appearing adjacent mucosa. Histological features include glandular crypt abnormalities, fibromuscular obliteration of the lamina propria, and thickened and splayed muscularis mucosa. CONCLUSIONS: Prolapsing mucosal polyps of the colon are histologically similar to other mucosal prolapsing conditions in the GI tract, such as the solitary rectal ulcer syndrome, inflammatory cloacogenic polyps, inflammatory "cap" polyps, and gastric antral vascular ectasia, and should therefore be designated as part of the "mucosal prolapse syndrome." PMID- 11866276 TI - Alterations in pulmonary function in inflammatory bowel disease are frequent and persist during remission. AB - OBJECTIVES: Information on the occurrence and frequency of pulmonary involvement in patients with inflammatory bowel disease (IBD) is inconsistent. The aim of this prospective study was to determine the frequency and type of pulmonary dysfunction in patients with IBD. METHODS: Sixty-six patients with IBD (35 with Crohn's disease [CD] and 31 with ulcerative colitis [UC]) and 30 control patients were investigated with respect to the following pulmonary function tests: forced expiratory volume in 1 s (FEV1), inspiratory vital capacity (IVC), Tiffeneau value (FEV1/IVC), and lung CO transfer capacity (D(LCO)). Disease activity in IBD patients was assessed by the CD activity index for CD and the Truelove index for UC, respectively. Smoking habits and medication were documented in every patient. RESULTS: Fourteen of 36 CD patients (39%) and 14 of 31 UC patients (45%) but only one of the controls exhibited at least one pathological (<80% of predicted value) pulmonary function test. In both CD and UC lung function tests were significantly decreased in comparison to the control group. This could be shown for FEV1 (-14% of predicted value in CD and -17% in UC, p < 0.01), IVC (-10% in CD and -12% in UC, p < 0.05), and DLCO (-20% in CD and -31% in UC, p < 0.01) without significant differences between both disease entities. The impairment of pulmonary function tests was more pronounced in patients with active disease than in those with inactive disease (FEV1, 81.4% vs 93.4% predicted, p < 0.02; IVC, 84.4% vs 93.7%, p < 0.05; DLCO, 80.4% vs 95.8%, ns). CONCLUSIONS: IBD patients show significantly decreased lung function tests in comparison to healthy controls. The impairment in active disease exceeded that during remission. PMID- 11866277 TI - Complementary medicine use in children and young adults with inflammatory bowel disease. AB - OBJECTIVES: We examined the use of complementary alternative medicine (CAM) in children and young adults with inflammatory bowel disease. METHODS: After validation of a questionnaire and completion of a pilot survey, children and young adults with inflammatory bowel disease were enrolled in three centers of pediatric gastroenterology (Boston, Detroit, and London). RESULTS: Two hundred eight questionnaires were completed in total (Boston, 120; Detroit, 37; London, 51). Ages ranged from 3.8 to 23.0 yr, 58% were male, 57% had Crohn's disease, and 35% had ulcerative colitis. The frequency of CAM use was 41%. The most common CAMs were megavitamin therapy (19%), dietary supplements (17%), and herbal medicine (14%). Parental CAM use and the number of adverse effects from conventional medicines were predictors of CAM use (odds ratio = 1.9, 95% CI = 1.2 3.1, p = 0.02; odds ratio = 1.3, 95% CI = 1.2-1.5, p < 0.001, respectively). The most important reasons respondents gave for using CAM were side effects from prescribed medicines, prescribed medicines not working as well as they had hoped, and hoping for a cure. Fifty-nine percent of respondents not taking CAM were interested in learning more about it. CONCLUSIONS: In our survey over 40% of children with chronic inflammatory bowel disease used complementary medicine in addition to conventional therapies. Parental CAM use and number of adverse effects from conventional therapies were the only independent predictors of CAM use. Some complementary therapies have potential for adverse effects and for drug interactions with conventional treatments. Physicians should take a thorough history of CAM use in children with chronic inflammatory bowel disease. PMID- 11866278 TI - Quality of life in inflammatory bowel disease in remission: the impact of IBS like symptoms and associated psychological factors. AB - OBJECTIVES: Quality of life is reduced in inflammatory bowel disease (IBD). Whether or not this is true in IBD patients in long-standing remission is unclear. Symptoms compatible with irritable bowel syndrome (IBS) are common in IBD patients in remission. The importance of psychological factors in this process is a matter of controversy. METHODS: Forty-three patients with ulcerative colitis (UC) and 40 with Crohn's disease (CD), who had been in remission for at least 1 yr according to laboratory parameters and clinical and endoscopical appearance, were included. These patients completed four different self administered questionnaires, evaluating GI symptoms, anxiety, depression, and psychological general well-being. The two patient groups were compared with the general population, and within-group comparisons in psychometric scores were made between patients with and without IBS-like symptoms. RESULTS: The psychological well-being in IBD patients in long-standing remission was similar to that of the general population, despite the presence of more severe GI symptoms. CD patients reported more psychosocial dysfunction, reduced well-being, and GI symptoms than UC patients. Thirty-three percent of UC patients and 57% of CD patients had IBS like symptoms. The group with IBS-like symptoms (both UC and CD) had higher levels of anxiety and depression and more reduced well-being than those without. Anxiety and reduced vitality were found to be independent predictors for IBS-like symptoms in these patients. CONCLUSION: The prevalence of IBS-like symptoms in IBD patients in long-standing remission is two to three times higher than that in the normal population. Psychological factors seem to be of importance in this process. However, as a group IBD patients in remission demonstrate psychological well-being comparable to that of the general population. PMID- 11866279 TI - Diagnosis of liver nodules observed in chronic liver disease patients during ultrasound screening for early detection of hepatocellular carcinoma. AB - OBJECTIVES: The aim of our study was to evaluate the nature of focal liver lesions detected during the ultrasound follow-up of a population (prevalently anti-hepatitis C virus [anti-HCV] positive) with chronic liver disease. METHODS: The study population consisted of 1827 consecutive newly diagnosed chronic liver disease cases without liver nodules at enrollment. Patients were screened at 4 month intervals by ultrasound and serum alpha-fetoprotein assessment. All lesions detected on imaging studies (except those accompanied by diagnostic a-fetoprotein levels) were subjected to biopsy (histology and cytology). RESULTS: During the 7 yr follow-up period (mean = 43.1 months), one or more solid focal lesions were found in 287 patients. a-Fetoprotein was diagnostic for hepatocellular carcinoma in 51 patients. Ultrasound-guided fine-needle biopsy was performed in the remaining 236 patients, yielding a diagnosis in 214: 198 hepatocellular carcinomas, 11 dysplastic nodules, and five B-cell non-Hodgkin's lymphomas (all confined to the liver and all in patients with chronic HCV infection). Twenty-two patients with nondiagnostic biopsies received diagnoses of hepatocellular carcinoma (20) or dysplastic nodules (two) based on arteriography or surgical biopsy. CONCLUSIONS: Focal lesions arising in patients with HCV-related chronic liver disease can be other than hepatocellular carcinoma, and ultrasound-guided fine-needle biopsy plays an important role in their diagnosis. The prevalence of non-Hodgkin's lymphoma in this selected population was 0.31%. The fact that all five lymphoma patients had cirrhosis related to hepatitis C strengthens the hypothesis of an etiological correlation between the latter infection and B-cell lymphoproliferative disorders. PMID- 11866280 TI - Viral genotype and hepatitis B virus DNA levels are correlated with histological liver damage in HBeAg-negative chronic hepatitis B virus infection. AB - OBJECTIVES: We aimed to study the relationship between the hepatitis B virus (HBV) genotypes, core promoter/precore stop codon mutations, and histological liver damage among hepatitis B e antigen (HBeAg)-negative patients. METHODS: Liver biopsy specimens of 55 HBeAg-negative chronic HBV-infected patients were studied. A histological activity index was scored for degree of necroinflammation (HAI-NI) and fibrosis (HAI-F) as described by Knodell et al. HBV DNA was determined by a cross-linking assay and polymerase chain reaction (PCR) at the core promoter/precore region and the S region. PCR-positive samples were directly sequenced for core promoter and precore mutations and examined by restriction fragment length polymorphism for genotyping. RESULTS: Forty-one males and 14 females at a median age of 43 were studied. HBV DNA was detectable in 32 (58%) and 37 (67%) patients by the cross-linking assay and PCR, respectively, at the time of liver biopsy. The median (range) HAI-NI and HAI-F scores were 5 (1-10) and 2 (0-4), respectively. HBV DNA detectable by either the cross-linking assay or PCR was associated with a higher HAI-NI score. Eleven and 31 patients had genotypes B and C HBV, respectively. Genotype C HBV was associated with higher HAI-NI than genotype B HBV. Core promoter mutations and precore stop codon mutation were detected in 74% and 40% patients, respectively, but they were not associated with higher HAI-NI or HAI-F scores. CONCLUSIONS: Detectable HBV DNA and genotype C HBV, but not core promoter or precore stop codon mutations, are associated with more severe liver damage in HBeAg-negative patients. PMID- 11866281 TI - Antibodies to soluble liver antigen/liver pancreas and HLA risk factors for type 1 autoimmune hepatitis. AB - OBJECTIVE: Antibodies to soluble liver antigen/liver-pancreas are highly specific markers of type 1 autoimmune hepatitis that have been associated with relapse. Our aim was to determine if these antibodies are reflective of a genetic predisposition for recrudescent disease. METHODS: One hundred forty-four white North American patients were evaluated by an enzyme immunoassay and by Western blot using recombinant soluble liver antigen/liver-pancreas; 122 were assessed for class II human leukocyte antigens (HLAs). RESULTS: Twenty-two patients (15%) had antibodies to soluble liver antigen/liver-pancreas. These patients were indistinguishable from seronegative patients by clinical, laboratory, and histological features at presentation. Patients with antibodies to soluble liver antigen/liver pancreas had HLA DR3 (79% vs 50%, p = 0.02) more commonly and HLA DR4 less often (16% vs 47%, p = 0.02) than patients with smooth muscle antibodies and/or antinuclear antibodies. Seropositivity was associated with DRB1*0301 and seronegativity was associated with DRB1*0401. Relapse after drug withdrawal occurred in all patients with antibodies to soluble liver antigen/liver-pancreas and at a higher frequency than in patients with conventional antibodies (100% vs 78%, p = 0.05). CONCLUSIONS: Antibodies to soluble liver antigen/liver pancreas are associated with HLA DR3 and the susceptibility allele, DRB1*0301. Antibodies to soluble liver antigen/liver-pancreas may be surrogate markers of a genetic propensity for recrudescent disease or the target autoantigen. They may be complementary to antinuclear antibodies and smooth muscle antibodies in diagnosis and management. PMID- 11866282 TI - Changes in serum levels of hepatitis C virus genotype 1b monitored by real-time quantitative polymerase chain reaction as a predictor of long term response to interferon-alpha treatment. AB - OBJECTIVE: The aim of this study was to find whether there is a relationship between the changes in the amounts of hepatitis C virus (HCV) at the start of interferon treatment and the long term response to therapy. METHODS: In 20 patients with HCV genotype 1b each given 880 MU of interferon-alpha, the changes in serum HCV RNA during the first 2 wk of therapy were monitored by real-time quantitative polymerase chain reaction (PCR). RESULTS: Real-time quantitative PCR detected HCV RNA at 10(1)-10(8) copies/ml. Serum HCV RNA decreased rapidly between 8 and 24 h after the first administration (first phase) and more slowly thereafter (second phase), with median exponential decays of 2.14 and 0.11 log10/day, respectively. Four patients had sustained virological responses, nine patients had transient responses, and seven patients had no responses. The differences in the rate of first-phase viral decline among the three groups were not significant (p = 0.34), but the differences in the rate of second-phase viral decline were significant (p = 0.0004); the median viral decline (interquartile range) in the second phase was 0.48 (0.42-0.50) log10/day in patients with sustained responses, 0.16 (0.10-0.19) log10/day in patients with transient responses, and 0.026 (0.017-0.040) log10/day in patients with no responses. CONCLUSIONS: Changes in serum levels of HCV genotype 1b in the first 2 wk of interferon-alpha treatment, monitored by real-time quantitative PCR, can be used for prediction of the long term therapeutic response. PMID- 11866283 TI - The cost-effectiveness of vaccinating chronic hepatitis C patients against hepatitis A. AB - OBJECTIVES: Although hepatitis A vaccination is recommended for persons with chronic liver disease, the cost-effectiveness of vaccinating patients with chronic hepatitis C virus has not been extensively studied. We evaluated its costs and benefits. METHODS: A Markov model was used to assess cost-effectiveness from the health system and societal perspectives. Costs of hepatitis A screening and vaccination were compared with savings from reduced hepatitis A treatment and work loss to determine net costs of a "screen and vaccinate" strategy. Net costs were compared with longevity gains to assess cost-effectiveness. RESULTS: Based on hypothetical cohorts of 100,000 patients, vaccination would reduce the number of hepatitis A cases 63-72%, depending on patient age. Screening and vaccination costs of $5.2 million would be partially offset by $1.5-$2.8 million reductions in hepatitis A treatment costs and $0.2-$1.0 million reductions in work loss costs. From the health system perspective, vaccination would cost $22,256, $50,391, and $102,064 per life-year saved for patients vaccinated at ages 30, 45, and 60 yr, respectively. Cost-effectiveness ratios improve when work loss prevention is considered. Results are most sensitive to hepatitis A infection and hospitalization rates, and the rate used to discount future benefits to their present values. CONCLUSIONS: Hepatitis A vaccination of chronic hepatitis C patients would substantially reduce morbidity and mortality in all age groups examined. Consistent with other medical interventions for chronic hepatitis C patients, cost-effectiveness is most favorable for younger patients. PMID- 11866284 TI - The effect of high dose and short interval HBV vaccination in individuals with chronic hepatitis C. AB - OBJECTIVES: The efficacy of the standard hepatitis B virus (HBV) vaccination schedule in individuals with chronic hepatitis C is reported to be reduced. Our aim was to assess the response rate to high dose, short interval HBV vaccination in such individuals. METHODS: A total of 152 individuals with chronic hepatitis C were vaccinated with 40 microg of vaccine administered monthly for 3 months. Twenty-six individuals with no evidence of liver disease underwent the same vaccination schedule and were considered to be the control group. Hepatitis C virus (HCV)-positive subjects who did not seroconvert to anti-hepatitis B surface positivity after the third dose of the vaccine (nonresponder) were vaccinated with a fourth dose of vaccine (booster dose, 80 microg). RESULTS: One hundred nine of the 152 individuals with chronic hepatitis C (72%) seroconverted to anti hepatitis B surface positivity (> 10 mIU/ml), as compared to 24 of the 26 controls (92%, p < 0.05). Although individuals with chronic hepatitis C responded less frequently to high dose, short interval HBV vaccination than did the controls, no differences in terms of effective immunity (>100 mIU/ml) were evident among the two groups of responders (51% vs 54%). Also, no difference in response was reported between individuals with chronic active hepatitis C and controls (92% vs 80%). The response rate was significantly lower in cirrhotics than in the noncirrhotic group (54% vs 80%, p < 0.001). Besides cirrhosis, no other demographic or bioclinical factor was found to influence the response to vaccination. After the additional booster dose, the overall response was increased to 74% of the cirrhotics and 88% of the noncirrhotics. No major HBV vaccine-related adverse effects were seen. CONCLUSIONS: A high dose, short interval HBV vaccination schedule is safe in individuals with chronic hepatitis C. From these data, it is suggested that a high dose and a short interval between HBV vaccinations may produce an effective and early antibody response in such patients. PMID- 11866285 TI - Lifetime cancer-attributable cost of care for long term survivors of colorectal cancer. AB - OBJECTIVES: We aimed to determine cancer-related medical care costs for long term survivors of colorectal cancer. METHODS: The SEER-Medicare database was used to measure lifetime cancer-attributable costs of care for those with colorectal cancer surviving at least 5 yr versus age- and gender-matched controls. Costs were directly estimated, stratified by age at diagnosis and stage at diagnosis, for years 6-11 after diagnosis and then modeled to estimate lifetime costs. Cost differences between cancer cases and controls were compared to expected costs based on published guidelines for postcancer surveillance. RESULTS: Lifetime medical costs for long term survivors (future years not discounted) were up to $19,516 higher than control costs, and were highest for younger age groups and those with early-stage disease. Excess costs for cancer survivors exceeded expected surveillance costs by $2,223-8,822 for years 6-10 from the date of initial diagnosis. CONCLUSIONS: Cancer-attributable medical costs can be substantial for long term survivors, and exceed expected costs of surveillance. Future research is need to determine the components of excess cost in this survivor group. PMID- 11866286 TI - A population-based, community estimate of total colon examination: the impact on compliance with screening for colorectal cancer. AB - OBJECTIVE: Colorectal cancer screening is underutilized. Total colon examination (TCE), such as with colonoscopy, can have a significant effect on the measured compliance with screening, as colonoscopy may be able to be performed as infrequently as once every 10 yr. In a population-based survey we determined the prevalence and validated the self-reporting of TCE and assessed its impact on compliance with screening. METHODS: We interviewed an age- and sex-stratified random sample of 50- to 79-yr-old residents in two communities in southwestern Pennsylvania. Subjects reported ever having had and duration since last use of fecal occult blood testing (FOBT), flexible sigmoidoscopy (FS), rigid proctoscopy, barium enema, and colonoscopy. Self-reports of colorectal testing were validated via retrieval of procedure reports. RESULTS: Out of 1223 individuals sampled, 496 completed a telephone interview (40.6% overall and 58.3% of eligible contacts). In those without personal or family histories of colorectal cancer or personal histories of polyps (n = 377), 50%, 19.6%, 39.8%, and 17.5% reported ever having had FOBT, FS, barium enema, and colonoscopy, respectively. Thirty-one percent reported having FOBT within the previous year or FS within the previous 5 yr. Including TCE within the previous 5 yr increased the measured compliance to 39.7%. Compliance was significantly greater among subjects with family histories of colorectal cancer (62.9% vs 39.7%, odds ratio = 2.6, 95% CI = 1.3-5.2). Self-reports of recent colonoscopy were verified in 29 of 35 instances (83%). CONCLUSION: The prevalence of TCE in this population was significant, and including TCE substantially increased measured compliance with colorectal cancer screening. Self-reported use of colonoscopy was validated as accurate. PMID- 11866287 TI - A cost analysis of endoscopic ultrasound in the evaluation of esophageal cancer. AB - OBJECTIVE: The use of endoscopic ultrasound (EUS) with guided fine needle aspiration (FNA) of suspicious lymph nodes has become an important aid in the staging of esophageal carcinoma. The economic impact of this staging strategy has not yet been described. We applied a decision analysis model to compare the costs of EUS FNA, CT-guided FNA, and surgery in the management of esophageal tumors. A cost-minimization approach was employed, as viewed from the perspective of the payer. METHODS: A decision analysis model with three management arms was designed using DATA 3.5 software, taking the entry criteria as esophageal carcinoma without evidence of distant metastases as determined by CT. Detection of tumor on celiac lymph node (CLN) FNA signified unresectability and prompted palliative treatment: chemoradiotherapy with endoscopic esophageal stenting rather than surgery. Baseline probabilities were varied through plausible ranges using sensitivity analysis. Cost inputs were based on Medicare professional fees plus Medicare facility fees. The endpoint was the cost of management per patient. RESULTS: EUS FNA was the least costly strategy ($13,811), compared to CT FNA ($14,350) and surgery ($13,992). The model outcome was sensitive to changes in both EUS FNA sensitivity and prevalence of CLN metastases. EUS FNA remained the least costly option provided the prevalence of CLN involvement was >16%; below this value, surgery became the most economical strategy. CONCLUSION: By minimizing unnecessary surgery, primarily by detecting CLN involvement, EUS FNA is the least costly staging strategy in the workup of patients with nonmetastatic esophageal cancer. Under certain circumstances, surgery is the preferred strategy. PMID- 11866288 TI - Gastroenterology training and career choices: a prospective longitudinal study of the impact of gender and of managed care. AB - OBJECTIVE: We aimed to determine if gender differences exist in the selection and training of female and male gastroenterology fellows. METHODS: One hundred seventy-six of 218 training program directors returned an 18-question survey about their programs, including leave policies, training, and prevalence of female faculty. Two cohorts of graduating trainees from 1993 and 1995 (N = 393) returned anonymous surveys regarding their training program experiences, demographics, and business training. RESULTS: Female gastroenterology trainees are more likely to choose programs according to parental leave policies (p < 0.05), female faculty (0.2990 correlation coefficient), and "family reasons" (p < 0.04) than the male trainees. Female trainees were more likely to remain childless (p < 0.001) or have fewer children at the end of training despite marital status not unlike their male colleagues. Female trainees altered their family planning because of training program restrictions (20% vs 7%, p < 0.001). They perceived gender discrimination (39%) and sexual harassment (19%) during gastroenterology training. Trainees of both sexes had mentorship during training (65% vs 71%, ns); female trainees were more likely to have an opposite sex mentor (71% vs 3.4%) despite an almost 50% prevalence of female full-time and clinical faculty. Female trainees were apt to be less trained in advanced endoscopy (p < 0.005). Trainees of both sexes were influenced by the changing health care environment in career choice (49% vs 42%, ns); neither gender felt adequately prepared for the business aspects of gastroenterology. CONCLUSION: Alterations in gastroenterology training are needed to attract qualified female applicants. New graduates of both sexes lack practice management education. PMID- 11866289 TI - The otoscope. PMID- 11866290 TI - Controlled transperitoneal percutaneous cecostomy as a therapeutic alternative to the endoscopic decompression for Ogilvie's syndrome. AB - Acute colonic pseudo-obstruction, the so-called Ogilvie's syndrome, results in massive colonic dilation without mechanical obstruction. In most cases, a conservative treatment with or without endoscopic decompression is sufficient. In rare cases of relapses or failures, a cecostomy has to be performed. A surgical cecostomy is associated with high morbidity and mortality. However, a percutaneous cecostomy could be an interesting alternative treatment. We report the case of a 67-yr-old male with colonic pseudoobstruction for which both the conservative and the endoscopic treatments were unsuccessful. A percutaneous cecostomy was performed, and for the first time in this indication, a transperitoneal access was used with the help of nylon T-fasteners. PMID- 11866291 TI - A case of severe benign intrahepatic cholestasis treated with liver transplantation. AB - A male patient with benign recurrent cholestasis since age 2.5 yr developed unremitting cholestasis with incapacitating pruritus and hepatic fibrosis by age 21. He was tried on numerous medical therapies for pruritus with transient or no relief. He responded only temporarily to biweekly plasmapheresis, which was carried out for 4 yr. He underwent orthotopic liver transplantation at age 25 with immediate resolution of his pruritus. At age 30 he is a happy, asymptomatic, fully employed professional. PMID- 11866292 TI - Rapidly progressive fibrosing cholestatic hepatitis--hepatitis C virus in HIV coinfection. AB - Fibrosing cholestatic hepatitis (FCH) is a severe and progressive form of liver dysfunction seen in organ transplant recipients infected with hepatitis B virus or hepatitis C virus (HCV) and has been attributed to cytopathic liver injury. To date, no case of FCH due to HCV has been reported in HIV-positive individuals. We describe two cases of HCV-induced FCH in two patients coinfected with HIV, culminating in rapidly progressive liver failure and death. Histological features and progression in both cases were not consistent with drug effect or obstruction. Late institution of interferon-based therapy was ultimately unsuccessful. The HCV RNA was not markedly elevated in these cases, suggesting that the cytopathic effect of HCV in these patients was not simply a consequence of viral load. FCH may in part explain the accelerated development of cirrhosis previously observed among coinfected patients. Clinicians should remain vigilant for FCH in the HIV/HCV population and consider antiviral treatment in this setting. PMID- 11866293 TI - P-ANCA for ulcerative colitis management-hype or hope? PMID- 11866294 TI - Liver injury during gastroenteritis in adults: a retrospective study in 727 cases. PMID- 11866295 TI - Irregular regeneration of hepatocytes and development of hepatocellular carcinoma in primary biliary cirrhosis. PMID- 11866296 TI - Real-time measurement of anti-HBs level and donor-specific transfusion via portal vein may reduce amount of HBIG after living related liver transplantation. PMID- 11866297 TI - Clarithromycin use preceding fulminant hepatic failure. PMID- 11866298 TI - Genetic and nutritional predictors of hyperhomocysteinemia in inflammatory bowel disease. PMID- 11866299 TI - Diagnosis of chronic liver disease: reproducibility and validation of liver biopsy. PMID- 11866300 TI - Three times weekly versus daily dose alpha-interferon treatment in patients with acute hepatitis C. PMID- 11866301 TI - Lamivudine and rapid regeneration of the atrophic liver in decompensated cirrhosis due to hepatitis B. PMID- 11866302 TI - Hyperinsulinemia in nondiabetic, both obese and nonobese patients with nonalcoholic steatohepatitis. PMID- 11866303 TI - Use of midazolam and propofol during colonoscopy: 7 years of experience. PMID- 11866304 TI - 24 weeks of interferon-ribavirin therapy should not routinely be given to all hepatitis C patients. PMID- 11866305 TI - Resolution of collagenous colitis after treatment for Helicobacter pylori. PMID- 11866306 TI - Treatment of acute uveitis associated with Crohn's disease and sacroileitis with infliximab. PMID- 11866307 TI - Three-dimensional ultrasound cholangiography: a new noninvasive technique for evaluation of biliary obstruction. PMID- 11866308 TI - Pioglitazone-associated fulminant hepatic failure. PMID- 11866309 TI - In support of "step-up therapy" for gastroesophageal reflux disease. PMID- 11866310 TI - Rectovesical ectopic varix intraperitoneal hemorrhage with fatal outcome. PMID- 11866311 TI - Acute pancreatitis during primary HIV-1 infection. PMID- 11866312 TI - Pemphigus vulgaris and ulcerative colitis. PMID- 11866313 TI - Mucosal IgA defect in primary biliary cirrhosis. PMID- 11866314 TI - Enteric and disseminated Campylobacter species infection during HIV disease: a persisting but significantly modified association in the HAART era. PMID- 11866315 TI - The clinical epidemiology of hysteria: vanishingly rare, or just vanishing? PMID- 11866316 TI - Sex differences in genetic and environmental risk factors for irrational fears and phobias. AB - BACKGROUND: For irrational fears and their associated phobias, epidemiological studies suggest sex differences in prevalence and twin studies report significant genetic effects. How does sex impact on the familial transmission of liability to fears and phobias? METHODS: In personal interviews with over 3000 complete pairs (of whom 1058 were opposite-sex dizygotic pairs), ascertained from a population based registry, we assessed the lifetime prevalence of five phobias and their associated irrational fears analysed using a multiple threshold model. Twin resemblance was assessed by polychoric correlations and biometrical model-fitting incorporating sex-specific effects. RESULTS: For agoraphobia, situational and blood/injury fear/phobia, the best fit model suggested equal heritability in males and females and genetic correlations between the sexes of less than +0.50. For animal fear/phobias by contrast, the best fit model suggested equal heritability in males and females and a genetic correlation of unity. No evidence was found for an impact of family environment on liability to these fears or phobias. For social phobias, twin resemblance in males was explained by genetic factors and in females by familial-environmental factors. CONCLUSION: The impact of sex on genetic risk may differ meaningfully across phobia subtypes. Sex specific genetic risk factors may exist for agoraphobia, social, situational and blood-injury phobias but not for animal fear/phobia. These results should be interpreted in the context of the limited power of twin studies, even with large sample sizes, to resolve sex-specific genetic effects. PMID- 11866317 TI - Psychiatric, demographic and personality characteristics of elderly sex offenders. AB - BACKGROUND: Psychiatric disorders are purported to play a role in the aetiology of violent crime, but evidence for their role in sexual offending is less clear. The authors investigated the prevalence of psychiatric morbidity and personality disorders in elderly incarcerated sex offenders compared with elderly non-sex offenders. METHOD: One hundred and one sex offenders and 102 non-sex offenders aged over 59 years wereinterviewed using standardized semi-structured interviews for psychiatric illness (the Geriatric Mental State) and the personality disorder (Structured Clinical Interview for DSM-IV personality disorders). Data on demographic, offence and victim characteristics were collected. RESULTS: Six per cent of the elderly sex offenders had a psychotic illness, 7% a DSM-IV major depressive episode and 33% a personality disorder; and 1% had dementia. These prevalence figures were not different from the elderly non-sex offenders interviewed in this study. Differences emerged at the level of personality traits with sex offenders having more schizoid, obsessive-compulsive, and avoidant traits, and fewer antisocial traits compared with non-sex offenders. CONCLUSIONS: Elderly sex offenders and non-sex-offenders have similar prevalence rates of mental illness. However, elderly sex offenders have increased schizoid, obsessive compulsive, and avoidant personality traits, supporting the view that sex offending in the elderly is associated more with personality factors than mental illness or organic brain disease. PMID- 11866318 TI - Mortality in psychiatric patients 5 to 21 years after hospital admission in Italy. AB - BACKGROUND: Mental disorders have an increased mortality risk. However, most data have been provided by few countries, some mental disorders have received little attention, long-term studies of large samples are scarce, and insufficient control for confounding variables has lead to artefactual inconsistencies across studies. The aims of this study were: to quantify the mortality risk in psychiatric patients 5 to 21 years after hospital admission and to investigate temporal trends in mortality risk and predictive factors associated with mortality. METHOD: All patients admitted to an in-patient psychiatric unit in Italy between 1978 and 1994 were included and vital status and death causes were determined up to 21 years after admission. The observed number of deaths in the sample was compared with the expected number of deaths in the general population. Cox proportional hazard models were fitted to identify predictors of mortality. RESULTS: Mortality from natural and unnatural causes was higher than expected across all mental disorders. Standardized mortality risk was higher in males (SMR = 4.55; 95% CI 4.17-4.97) than in females (SMR = 3.43; 95% CI 3.07-3.83). Individuals aged less than 40 years were at higher risk in both sexes. The first several years following admission were characterized by a faster decline in survival. Several demographic and clinical factors were predictors of mortality. CONCLUSIONS: Mortality is high in individuals with mental disorders. Prevention of unnatural death causes is an important goal though insufficient to abate excess mortality, since natural death causes account for it to a larger extent. PMID- 11866319 TI - Suicide in female nurses in England and Wales. AB - BACKGROUND: Female nurses appear to have an increased risk of suicide but the reasons are unknown. METHOD: We have concluded a study of nurse suicides (N = 106) in England and Wales, including a psychological autopsy study (N = 42) and case-control comparison with living nurses (N = 84). RESULTS: Nearly three quarters of the nurse suicides had previous contact with psychiatric services and almost half had been psychiatric in-patients in the past. There were particularly marked differences between the cases and controls for current psychiatric disorder (90.5% v. 7.1%, OR = 68.5), personality disorder (38.1% v. 12%, OR = 32), and history of deliberate self-harm (71.4% v. 2.4%, OR = 58.5). Family background and social factors (especially concerning interpersonal relationships) also distinguished the two groups. Smoking and serious alcohol abuse were much more frequent in the suicides. There was some indication that while many of the suicides were in contact with psychiatric services, care may not have been optimal in some cases. CONCLUSIONS: The most important strategies for suicide prevention in nurses are in prevention, detection and management of psychiatric disorders. In assessing suicide risk a history of DSH and the presence of comorbid psychiatric and personality disorders are particularly important. PMID- 11866320 TI - Recollection memory deficits in patients with major depressive disorder predicted by past depressions but not current mood state or treatment status. AB - BACKGROUND: Neuropsychological studies have suggested that memory systems reliant on medial temporal lobe structures are impaired in patients with depression. There is less data regarding whether this impairment is specific to recollection memory systems, and whether clinical features predict impairment. This study sought to address these issues. METHOD: A computerized process-dissociation memory task was utilized to dissociate recollection and habit memory in 40 patients with past or current major depression and 40 age, sex and IQ matched non psychiatric control subjects. The Cognitive Failures Questionnaire was used to assess patients' perceptions of day-to-day memory failures. RESULTS: Patients had impaired recollection memory (t = 4.7, P < 0.001), but no impairment in habit memory when compared to controls. Recollection memory performance was not predicted by indices of current mood state, but was predicted by self-assessments of impairment (beta = -0.33; P = 0.008) and past number of depressions (beta = 0.41; P = 0.001). There was no evidence that standard therapy with antidepressant medication either improved or worsened memory performance. CONCLUSIONS: The results confirm that patients with multiple past depressions have reduced function on recollection memory tasks, but not on habit memory performance. The memory deficits were independent of current mood state but related to past course of illness and significant enough that patients detected impairment in day-to-day memory function. PMID- 11866321 TI - Evidence of an early information processing speed deficit in unipolar major depression. AB - BACKGROUND: Slowing of the speed of information processing has been reported in geriatric depression, but it is not clear if the impairment is present in younger patients, if motor retardation is responsible, or if antidepressant medications play a role. METHOD: Twenty unmedicated unipolar depressed inpatients were compared with 19 medicated depressed in-patients and 20 age-, sex- and verbal IQ matched controls on inspection time (IT), a measure of speed of information processing that does not require a speeded motor response. We also examined the relationship between IT and current mood and length of depressive illness. RESULTS: Unmedicated depressed patients showed slowing of information processing speed when compared to both medicated depressed patients and controls. The latter two groups were not significantly different from each other. Slowing of IT was not associated with current mood, but was negatively correlated with length of illness since first episode. No differences in IT were found between patients receiving medication with anticholinergic effects and patients receiving medication with no anticholinergic effects. CONCLUSIONS: The findings indicate that unipolar depression is associated with a slowing of speed of information processing in younger patients who have not received antidepressant medication. This does not appear to be a result of motor slowing. PMID- 11866322 TI - Categoric overgeneral autobiographical memory in adolescents with major depressive disorder. AB - BACKGROUND: Categoric, overgeneral autobiographical memory is more common in depressed adults than controls and predicts persistence of depression. This cross sectional study investigated whether, compared with non-depressed psychiatric cases and community controls, first episode major depressive disorder (MDD) in adolescents is associated with categoric overgeneral memory retrieval. METHODS: Ninety-six clinically referred adolescents (aged 12-17 years) with MDD, 26 non depressed psychiatric cases and a sample of 33 community controls were recruited. All subjects were assessed using the Kiddie-Schedule for Schizophrenia and Affective Disorders, and completed Williams' cued Autobiographical Memory Test and the Mood and Feelings Questionnaire. Hamilton Depression Rating Scales were completed with MDD subjects, as an index of depression severity. RESULTS: Adolescents with current first episode MDD retrieved more categoric overgeneral memories than controls, but not than non-depressed psychiatric cases. Adolescents in full remission from a recent episode of MDD retrieved more categoric memories to positive cues than controls. Categoric memory in MDD was related to observer rated and self-reported severity, but not to the pattern of co-morbid diagnoses. There were negative correlations between IQ and categoric memories in both clinical cases and controls. A positive correlation between categoric memory to negative cues and self-reported depressive symptoms was found in clinical cases (but not controls). CONCLUSIONS: In adolescents, increased categoric overgeneral memory is associated with, but not specific to first episode MDD. Positive categoric memories are also increased in fully remitted MDD as compared to controls. PMID- 11866323 TI - Acts of control in schizophrenia: dissociating the components of inhibition. AB - BACKGROUND: Inhibitory deficits have been frequently reported in schizophrenia. Such deficits are usually associated with activities of prefrontal cortex and related networks. An understanding of intentional inhibitory control requires knowledge of how actions are planned and initiated and the components involved in stopping these actions. METHODS: Patients with schizophrenia, a psychosis comparison group and a healthy control group participated in a visual choice reaction time (go) task and attempted to inhibit their responses to the go task when an auditory 'stop' signal was heard. RESULTS: Schizophrenia patients demonstrated significantly slower response execution but the estimated speed of inhibition was not significantly different from that of healthy controls. Both patient groups were impaired in their ability to inhibit a response across a range of stop-signal delays. The poorer performance of schizophrenia patients only was related to a difficulty in reliably triggering the inhibitory response. CONCLUSIONS: Impaired response inhibition is not unique to schizophrenia. However, the nature of their problem is markedly different from that of other psychopathological groups. Possible neural mechanisms underpinning difficulties in triggering inhibitory responses and in the voluntary initiation of actions in schizophrenia are considered. PMID- 11866324 TI - Perceived need for mental health care: influences of diagnosis, demography and disability. AB - BACKGROUND: Recent major epidemiological studies have adopted increasingly multidimensional approaches to assessment. Several of these have included some assessment of perceived need for mental health care. The Australian National Survey of Mental Health and Wellbeing, conducted in 1997, included a particularly detailed examination of this construct, with an instrument with demonstrated reliability and validity. METHODS: A clustered probability sample of 10641 Australians responded to the field questionnaire for this survey, including questions on perceived need either where there had been service utilization, or where a disorder was detected by administration of sections of the Composite International Diagnostic Interview. The confidentialized unit record file generated from the survey was analysed for determinants of perceived need. RESULTS: Perceived need is increased in females, in people in the middle years of adulthood, and in those who have affective disorders or co-morbidity. Effects of diagnosis and disability can account for most of the differences in gender specific rates. With correction for these effects through regression, there is less perceived need for social interventions and possibly more for counselling in females; disability is confirmed as strongly positively associated with perceived need, as are the presence of affective disorders or co-morbidity. CONCLUSIONS: The findings of this study underscore the imperative for mental health services to be attentive and responsive to consumer perceived need. The substantial majority of people who are significantly disabled by mental health problems are among those who see themselves as having such needs. PMID- 11866325 TI - Combined effect of mental disorder and low social support on care service use for mental health problems in the Dutch general population. AB - BACKGROUND: People with a mental disorder have high rates of service utilization for emotional or addiction problems. Little is known about the role of functional impairments and low social support in such service use. This article investigates: (1) whether the presence of multiple functional impairments mediates the link between mental disorder and service use; and (2) whether social support modifies that association. METHODS: Data were derived from the Netherlands Mental Health Survey and Incidence Study, NEMESIS, a prospective general population study. Predictors of service use (mental disorder; functional impairments; social support) were recorded in the second wave of the study, and service use in the third wave. RESULTS: Persons with a DSM-III-R disorder and persons with multiple functional impairments were three to seven times as likely to use primary or mental health care. People with low perceived social support were two to three times as likely to use them, and living alone increased the likelihood by 30% to 80%. The effect of mental disorder on service use was mediated by multiple functional impairments. In people with a mental disorder, low levels of social support intensified mental health service use. CONCLUSION: Service utilization by people with mental problems can be better understood through a model incorporating: (1) independent effects of mental disorder, functional impairments and social support on service use; (2) a mediating effect of multiple functional impairments on the link between mental disorder and service use; and (3) interaction effects of mental disorder and low social support on service use. PMID- 11866326 TI - Personality and the perception of health in the general population. AB - BACKGROUND: Several population-based studies have shown that self-perceived health is a powerful predictor of health outcomes. The extent to which self perceived health is associated with personality characteristics is, however, largely unknown. We aimed to study the relationship between self-perceived health and personality among adults in the community. METHOD: Data were drawn from the Midlife Development in the United States Survey, a representative sample of adults age 25-74. MANOVA was used to determine the relationship between self perception of health and personality using the five-factor model. RESULTS: Personality factors were significantly associated with perception of poor health. Among those without self-reported medical problems (N = 834), openness to experience, extraversion and conscientiousness were associated with perception of good health, while neuroticism was associated with the perception of poor health. In subjects with self-reported medical problems (N = 2772), high scores on agreeableness, openness to experience, extraversion and conscientiousness, and low neuroticism scores were associated with perception of good health. These associations remained significant after adjustments for age, gender, race, marital status and education. CONCLUSIONS: Self-perceived health is strongly associated with personality characteristics, both in subjects with and without self-reported medical problems. It is suggested that personality characteristics could contribute to the previously reported associations between self-perceived health and health outcomes. PMID- 11866327 TI - Evidence that three dimensions of psychosis have a distribution in the general population. AB - BACKGROUND: The aims of the study were: first to examine, using clinical symptoms of patients as a template, whether the correlated but independent dimensions of positive, negative and depressive symptoms that have been identified in clinical psychosis, also have a distribution as non-clinical experiences in the general population; and second, to establish to what degree population variation in experience of positive and negative features of psychosis is actually independent of experience of depression. METHOD: In a representative population sample of 932 young men, we measured experiences of positive, negative and depressive features of psychosis, using a 40-item self-report instrument. Confirmatory factor analysis was used to compare the fit of hypothesized one-, two- and three-factor solutions. RESULTS: A three-factor model of separate depressive, positive and negative dimensions provided a better fit to the data than either a two-factor or unidimensional model. All three dimensions were correlated with each other, but also showed good discriminant validity in relation to established scales, confirming their relative independence. CONCLUSION: The data suggest that the correlated dimensions of clinical psychosis also have a distribution in the general population, and that depressive symptoms may form an integral part of psychosis-like experiences in the general population. PMID- 11866328 TI - Vascular risk factors in late onset mania. AB - BACKGROUND: Previous researches have suggested that late onset mania is a distinct subtype associated with medical and neurological disorders. Few studies, however, have focused on vascular risk factors. METHODS: Records of 366 bipolar patients were reviewed and age of first psychiatric hospitalization determined. Late-onset cases were determined empirically from a distribution histogram. Late onset cases were matched to early onset cases and histories of vascular disease/risks and current cholesterol levels compared. RESULTS: The distribution of age of first psychiatric hospitalization was bimodal with an intermode at age 47. Using that threshold, 6.3% of the cohort was classified as having late onset mania. Vascular risks factors were greater and current cholesterol levels higher in the late onset group. CONCLUSIONS: Late onset mania is associated with greater vascular risk factors. The bimodal appearance of age of first psychiatric hospitalization in this study provides further support of late onset mania as a distinct manic subtype with possibly a different, vascular aetiology. Control of these vascular risks may impact on the incidence of late onset mania, as well as on its clinical management. PMID- 11866329 TI - Spousal resemblance for history of major depressive episode in the previous year. AB - BACKGROUND: There is discrepancy in findings on spousal concordance for major depression. Here we report the risk of depression and its determinants in spouses of persons with or without depression, taking into account several known risk factors for major depression. METHODS: A random sample of non-institutionalized Finnish individual aged 15-75 years was interviewed in the 1996 National Health Care Survey. The sample included 1708 male-female spouse pairs. Major depressive episode (MDE) during the last 12 months was assessed using the Short Form of the University of Michigan version of the Composite International Diagnostic Interview (the UM-CIDI Short Form). Risk factors were assessed in the same interview. RESULTS: Factors associating with MDE were spouse's MDE, own alcohol intoxication at least once a week and own chronic medical conditions. In addition, there was a strong association between female's current smoking and male's MDE, independently of other risk factors and spousal MDE. The association of MDE with spouses's MDE was not affected by taking into account other assessed risk factors (own or spouse's). CONCLUSIONS: The results indicate elevated spouse concordance for MDE independent of the risk factors assessed in the present study. PMID- 11866330 TI - Procalcitonin: a marker of neonatal sepsis. AB - This study was designed to assess the value of procalcitonin in establishing the diagnosis and evaluating the prognosis of neonatal sepsis. Thirty-four infants with neonatal sepsis were included in the study. Procalcitonin values of the cases with sepsis were (2.21 +/- 2.48 ng/ml) significantly higher than the values in the control group (0.71 +/- 0.5 ng/ml; p = 0.01). On the 7th day of therapy neonates who had achieved clinical recovery had a significant decrease of procalcitonin levels (0.55 +/- 0.27 ng/ml) compared to the initial values (p = 0.001). Initial mean procalcitonin levels of the cases resulting in death were 4.31 +/- 3.66 g/ml. This was significantly higher than the initial values of the patients who had clinical recovery (1.18 +/- 1.24 ng/ml;p = 0.02). Procalcitonin is a valuable marker for diagnosis, for evaluating prognosis and response to therapy in neonatal sepsis. PMID- 11866331 TI - The characterization of bacterial isolates from acute otitis media in Ile-Ife, southwestern Nigeria. AB - The incidence of acute otitis media (AOM) in a comprehensive healthcare setting was investigated in Ile-Ife, Nigeria. Out of the 617 children examined, 53 (11.64 per cent) suffered from the condition based on the criteria used. Staphylococci constituted the predominant organisms associated with the condition with Staphylococcus aureus (25.0 per cent) being the most frequent single microbe recovered from the subjects. This was followed by Proteus mirabilis (16.2 per cent), Staphylococcus sp. (8.8 per cent), Streptococcus pneumoniae (8.8 per cent), Pseudomonas aeruginosa and Haemophilus influenzae (7.4 per cent each). Most isolates tested were multiply resistant to the antibiotics commonly employed in treating infections caused by these organisms. The study highlights the prevalence of multi-resistant organisms amongst the subjects and recommends prompt therapeutic intervention to avert ineffectiveness of antibiotics when used in treating infections caused by these organisms in the community. PMID- 11866332 TI - Targeting aid programmes on diseases that cause poverty. PMID- 11866333 TI - Predictive value of tuberculin induration at 24 h in healthy schoolchildren. AB - Tuberculin (1 TU PPD RT23 Tween 80) was administered to 500 healthy schoolchildren, aged 5-9 years. The induration at 24 h was compared with that at 72 h. The mean (standard deviation) induration of the tuberculin reaction at the end of 24,48 and 72 h was 4.5 (6.75), 4.4 (7.11) and 4.0 (6.91) mm, respectively. When the tuberculin reaction was 0-9 mm, a significant difference was noted between the 24- and 72-h reading (p = 0.0001). There was no difference in the size of the tuberculin reaction between the 24- and 72-h readings when the reaction size was > or = 10 mm (p > 0.05). Considering a tuberculin induration of < 10 mm as negative tuberculin reaction for our reference population in South India, the sensitivity, specificity, positive predictive value and the negative predictive value in our study for the 24-h reading were found to be high. The predictive value of 24-h induration was not affected by age. Reading and interpretation of tuberculin test after 24 h needs due consideration in clinical practice. PMID- 11866334 TI - Evaluation of serum soluble transferrin receptors and erythropoietin levels as indicators for erythropoietic activity among multi-transfused beta-thalassemic patients. AB - Eighty multi-transfused beta-thalassemics--40 of them on regular transfusion (RT) (every 3-4 weeks) and 40 irregularly transfused (IT)-and 20 age- and-sex-matched controls were evaluated for serum erythropoietin (sEpo) and soluble serum transferrin receptors (sTfr) as indicators of erythropoietic activity. All subjects were studied for pre-transfusion hemoglobin (Hb), reticulocytic index (RI), serum ferritin, sEpo and sTfr. Results showed that the mean RI, sEpo and sTfr values were significantly higher in the IT group (2.8; 80 mU/ml; and 19 microg/ml, respectively) compared to the RT group (1.2; 35.2 mU/ml; and 13.9 microg/ml, respectively) and controls (1.1; 22.6 mU/ml; and 7 microg/ml, respectively) (p < 0.05); while only the mean sTfr value was significantly higher in the RT group compared to controls. The mean pre-transfusion Hb was significantly lower in the IT group (8 g/dl) compared to both the RT group (9.3 g/dl) and controls (12.5 g/dl) (p < 0.05); while the RT group level was significantly lower than controls. There was an inverse correlation between Hb level on one hand and sEpo (r = 0.324,p < 0.05), sTfr (r = -0.651, p < 0.05) and RI (r = -0.451, p < 0.05) on the other. In summary, sEpo, sTfr and RI could be used as accurate and reliable indicators of successful erythroid marrow suppression and for the determination of optimal pre-transfusion Hb level in thalassemia on an individual basis, with sTfr being the most sensitive indicator. PMID- 11866335 TI - Association of gastroesophageal reflux disease in young children with persistent respiratory symptoms. AB - Forty children aged between 3 months and 3 years (median age 14 months) with persistent respiratory symptoms beyond 4 weeks or recurrence of respiratory symptoms were investigated for gastroesophageal reflux (GER). Diagnostic tests included upper gastrointestinal endoscopy, oesophageal biopsy, gastroesophageal scintiscan and 24 h ambulatory oesophageal pH monitoring. GER was detected in 14 (35 per cent) of these patients; which included 38 per cent of the enrolled cases of recurrent bronchopneumonia, 40 per cent cases of reactive airway disease, and 22 per cent cases of persistent cough. Amongst the cases detected to have GER, the age of onset of respiratory symptoms was less than 1 year in 86 per cent of cases (p < 0.01), nocturnal symptoms of cough and wheeze were reported in 78 per cent (p < 0.05), and 86 per cent cases did not present with typical gastrointestinal symptoms (p < 0.01). Family history of asthma was absent in all cases of GER-related reactive airway disease (p < 0.01). Cases detected to have GER were followed for 3-6 months after starting anti-reflux therapy. A significant (p < 0.01) decrease was noticed in the number of further episodes in children with GER-related recurrent bronchopneumonia and reactive airway disease after starting anti-reflux therapy. Improvement was also noticed in nocturnal symptoms and nutritional status after anti-reflux therapy was started. Our results suggest that GER may be one of the possible contributing factors in any child with recurrent and persistent respiratory complaints. Early diagnosis and anti-reflux therapy in cases with GER-related respiratory complaints can result in significant improvement in symptoms. PMID- 11866336 TI - Neonatal outcomes at a subdistrict hospital in north India. AB - Subdistrict hospitals form the first referral level facilities for essential newborn care services. However, there is a paucity of information on the pattern of neonatal admission and outcomes at this level. The objective of this study was to describe the spectrum of neonatal mortality at a subdistrict hospital. The study was conducted at a 50-bed hospital at Ballabgarh in northern India. The data of the neonates born in this hospital (inborns) and those admitted with sickness after being delivered at home (outborns) were separately analysed for the period 1994-1999. The main outcomes of interests were incidence, distribution, and primary causes of neonatal mortality. Of 6746 inborns and 385 outborns admitted (total 7,137) there were 56 deaths (0.8 per cent) and 38 (0.6 per cent) referrals among inborn and 70 deaths (18.2 per cent) and 37 (9.6 per cent) referrals among outborns. The deaths or referral rates among inborn for different weight groups were 27.7 per cent for < 1,500 g, 7.2 per cent for 1500 1999 g, 1.2 per cent for 2,000-2,499 g, and 0.6 per cent for weight > or = 2,500 g. Most deaths under 7 days of age were related to prematurity [41 per cent (28/69)] and birth asphyxia [38 per cent (26/68)], while those aged between 7 and 27 days were mostly due to sepsis [91 per cent (42/46)]. The results of this study indicate that babies with a birthweight above 1,500 g have a good outcome at this level. Deaths under 7 days of age were mostly due to birth asphyxia and prematurity, while those after 7 days were almost entirely due to sepsis. Referral is an important outcome at this level of service. PMID- 11866337 TI - CT and MR patterns of hypoxic ischemic brain damage following perinatal asphyxia. AB - The objectives were to study the clinical and neurological abnormalities in children with cerebral palsy and to attempt to correlate the signs with radiological abnormalities detected by CT scan and/or MRI of the brain. In a prospective, hospital-based study, 65 children with cerebral palsy were examined neurologically and their perinatal history was reviewed. Their cranial CT scan, and/or magnetic resonance images were studied. The association between the gestational ages, perinatal history, neurological deficits, and the radiological appearances were studied. Of the 24 preterm-born and 41 term-born children, 23 had spastic diplegia; 57 per cent of these children has significant periventricular leucomalacia, which was more common among preterm-born children. Of the 13 children with hemiplegia, 12 had unilateral lesions on neuroimaging. Spastic tetraplegia was associated with extensive, bilateral, diffuse brain damage. Extrapyramidal cerebral palsy was far more common among term-born infants and 80 per cent of these showed significant abnormalities in the basal ganglia region. Ataxic cerebral palsy was an uncommon variety and there was no significant correlation between neurological signs and abnormalities on brain imaging. In conclusion, the radiological findings were closely related to the type of cerebral palsy and the neurological deficit except in the ataxic type. We believe that CT and MRI imaging are helpful in understanding the pathology and the timing of the lesion in cerebral palsy. PMID- 11866338 TI - Endocrine complications in patients with beta-thalassemia major. AB - Thirty-seven patients with thalassemina major (TM) were studied to determine the extent and rate of endocrine complications. Mean haemoglobin and ferritin concentrations were 8.8 +/- 0.6 and 3,597 +/- 1,931, respectively. Provocation tests for growth hormone secretion were applied in patients with standing heights below the third centile and/or growth velocities below the 10th centile. Sexual maturation was assessed by using the criteria of Tanner. Glucose metabolism was assessed by fasting plasma glucose and glucose tolerance test. Basal thyroid function was measured and thyrotropin-releasing hormone tolerance test was carried out. Growth retardation was found in 40 per cent of patients and growth hormone deficiency was a prominent cause of growth retardation. Gonadal dysfunction was detected in 47 per cent of patients. Hypothyroidism was observed in 16 per cent and impaired glucose metabolism in 10.8 per cent patients. The high rate of endocrine disturbances indicates the importance of regular follow-up of thalassemia major patients with regard to endocrine complications of the disease. PMID- 11866339 TI - Cervical intramedullary tuberculoma: acute presentation and rapid response to medical therapy. AB - A case of an 11-year-old boy with rapidly progressing quadriparesis with bowel and bladder incontinence is reported. MRI of the spine revealed an intramedullary tuberculoma at the level of C5-7. Investigations further revealed evidence of tuberculosis in the brain and lungs as well. The child made a rapid recovery with medical management alone. PMID- 11866340 TI - Value of neuron-specific enolase levels in cerebrospinal fluid in evaluating the prognosis of asphyxiated neonates. PMID- 11866341 TI - Interaction of maize zein with wheat gluten in composite dough and bread as determined by confocal laser scanning microscopy. AB - Protein body-free maize zein, when mixed at 35 degrees C (above its glass transition temperature range), significantly (p < 0.01) improved the rheological and leavening properties of sorghum-wheat composite flour dough, resulting in improved loaf volume. Confocal laser scanning microscopy was used to observe the structure of zein fibrils and the interaction between zein and gluten proteins in the composite dough and bread systems. Autofluorescence and immunolocalization techniques were used to locate gluten and zein, respectively. Optical sections were collected every 0.4 microm through the samples and digitally processed to produce reconstructed three-dimensional images. Results showed that zein fibrils form an outer layer that intermittently coats the gluten networks, thereby strengthening them. This type of microstructure is able to withstand the pressure exerted by gas cell expansion during yeast fermentation to increase loaf volume. PMID- 11866342 TI - Development of preparation methods for and insights obtained from atomic force microscopy of fluid spaces in cortical bone. AB - Several preparation methods were developed to investigate the dimensions and surface structure of fluid spaces within cortical bone, using atomic force microscopy (AFM). Of special interest was the morphology of the lacunocanalicular system, which serves as a conduit between osteocytes encased in bone tissue, the intramedullary cavity, blood vessels running through the bone, and the periosteal surface of bone. Fracture and the removal of either the mineral or the organic component is a method by which each component can be investigated at a very high resolution in situ. Although fractured bone was too rough to image details of the lacunocanalicular system, post-treatment with ethylene diamine tetraacetic acid (EDTA) or papain allowed for investigation of the collagen matrix or the mineral crystals of bone, respectively. Cut and polished bone was smooth enough for identifying the lacunae of bone using AFM, but unambiguous differentiation between the canaliculi and cracks in the bone surface was not possible. However, when the lacunocanalicular system was filled with polymethylmethacrylate (PMMA), it was possible to image casts of the lacunocanalicular system by selectively etching away the surrounding bone matrix. Using this method, we identified individual canaliculi and measured their dimensions. Furthermore, by carefully etching away the bone matrix in successive etches, it was shown that the wall structure of the canaliculus is dominated by collagen fibrils. These observations have important implications for fluid flow in bone. PMID- 11866343 TI - A quantitative cryo-scanning X-ray microanalysis protocol for the examination of the eye. AB - Analysis of elements present in fluids contained in small, poorly accessible sections of biological tissue is challenging. The choroid of the eye, which is a vascular tissue approximately 100 microm thick, surrounds the retina for the purposes of nutrient supply and metabolite removal, and which in the chick shows dramatic volumetric change in response to visual experiences. Because fluid homeostasis is critical to good vision, a complete understanding of the ionic changes driving large shifts in ocular fluids is required. However, the structure of the choroid and retina make extraction of pure fluids for analysis extremely difficult. Elemental x-ray analysis on a transverse chorioretinal specimen was performed after rapid freezing of a whole chick eye in liquid nitrogen, and mechanically fracturing the frozen globe. Using a Polaron Cryotrans System on a Cambridge S-360 scanning electron microscope and a Kevex Quantum detector, spectra were obtained for blood vessels, lymphatic vessels and vitreous that were readily visible at 265x. Analysis was performed on a frozen control solution of the elements found in the vessels. The elements and their concentrations found in blood vessels by x-ray analysis compared well with those from whole blood as established by conventional means. The analysis for lymph yielded results compatible with expectations; no other published data for small lymphatics enable a direct comparison. In conclusion, x-ray analysis can be used to acquire information that is otherwise unobtainable from tissue in situ. The same bulk frozen elemental microanalysis protocol would have application to other organs and tissues when access to the site would destroy the integrity of the tissue under investigation. PMID- 11866344 TI - Line profile reconstruction from simultaneously recorded secondary and backscattered electron signals. AB - We propose a reconstruction method of surface morphology using a combination of secondary and back-scattered electron signals from the scanning electron microscope (SEM). Compared with multiple-detector methods, the proposed system requires only conventional secondary and backscattered electron detectors for a line profile reconstruction in one direction. This method is an application of genetic algorithms to the measurement of surface morphology in SEM. We use the chi-square distribution of the reconstruction error as the objective function within a scheme to minimize the number of vertices in the reconstructed surface profile. (The reconstruction error is the relative difference between the calculated and experimental data.) To evaluate the efficacy of our method, a surface profile is successfully reconstructed from a pair of line scans across the center of a latex particle. PMID- 11866345 TI - An empirical energy loss equation of electrons. AB - A modified Love-Cox-Scott (1978) equation of electron energy loss has been suggested. The stopping powers predicted by the modified Love-Cox-Scott equation are compared with those by the Tung et al. (1979) model, the Joy and Luo (1989) equation, and the experimental data given in database of Joy at: http://web.ukt. edu/-scrutk. In the energy range of E0< or = 5 keV, the Monte Carlo simulations of the electron scattering in Al, Ag, and Au have been performed, applying the Mott cross section for elastic scattering and the modified Love-Cox-Scott equation (1978) and the equations by Love et al. (1978) and Joy and Luo (1989), respectively, for the inelastic scattering. The calculated results on the backscattering coefficients, the energy distributions of the backscattered electrons, and the energy dissipation of the electron based on the three equations are compared. PMID- 11866346 TI - Convolution and correlation: a case study of scanning imaging and analysis systems. AB - The relationship between convolution/correlation operation and the data acquisition process of the scanning microscope and spectrometer families is analyzed. It is shown that a coordinate or event sensitive detector, and the intrinsic or extrinsic property of the specimen response, are two important factors in distinguishing the data acquisition mode of such systems. Four types of convolution- and correlation-based modes are extracted to illuminate the physical characteristics of scanning imaging and analysis systems by focusing on the probe, specimen, detector, and their relationships. Criteria for identifying these modes are explored. In addition, the physical meanings of general existing coefficients between the independent variables of convolution and correlation are investigated. PMID- 11866347 TI - Joining together. PMID- 11866348 TI - Vascular factors in carpal tunnel syndrome. AB - An increase in hand and forearm volume was induced without hand movement in ten subjects who had carpal tunnel syndrome (CTS). A tester, unaware of sides affected by CTS, performed the Volume Provocation Test (VPT) by inflating a sphygmomanometer cuff around the upper arm to 15 mm Hg less than diastolic pressure for four minutes. Pre- and post-test volumes, intensity of discomfort, and quality and distribution of produced symptoms were recorded. The VPT induced significant increases of segment volumes bilaterally (P1 tailed <0.05), but not more on the side of strongest symptoms compared to the less affected, or asymptomatic side (P1 tailed = 0.07). Mean discomfort on the side of strongest symptoms (5.4/10) was significantly higher (P1 tailed <0.01) than on the contralateral side (2.4/10). In 44% of the affected arms, some or all of the subjects' nocturnal symptoms were reproduced after volume increase, suggesting that CTS is a vascular phenomenon in these patients. PMID- 11866349 TI - A research agenda for hand therapy. AB - A survey, to which 401 hand therapists responded, was conducted to establish research priorities in the field of hand therapy and to investigate barriers to hand therapy research. Results indicated that reimbursement issues, particularly those generated by managed care, were reported to have adversely affected quality of patient care, job satisfaction, job security, and ability to conduct research. Research priorities were defined in this study, and the need for clinical trials to determine the effectiveness of hand therapy interventions was found to be urgent. Research questions identified by respondents reflected both the traditional disorders treated by hand therapists, such as tendon repairs, and a greater concern with issues concerning cumulative trauma. Barriers to research were reported by respondents, and ways in which professional organizations could meet clinicians' needs in conducting and utilizing research were highlighted. PMID- 11866350 TI - Compliance with hand therapy programs: therapists' and patients' perceptions. AB - AIM: This study explored patients' and hand therapists' perceptions of compliance with hand therapy and compared these perceptions. METHOD: Forty-one patients attending hand therapy at a large metropolitan hospital and 69 hand therapists, all members of the Australian Hand Therapy Association, were interviewed by telephone using a questionnaire developed for the study. Demographic data were analyzed descriptively and patients' and therapists' perceptions compared using chi-square. RESULTS: Patients' and therapists' perceptions differed at a statistically significant level (p < 0.01, adjusted alpha rate) for 24 of the 33 items measuring perceptions of compliance with hand therapy. CONCLUSIONS: Therapists and patients have differing perceptions of compliance. PMID- 11866352 TI - The effects of normal human variability and hand activity on sensory testing with the full Semmes-Weinstein monofilaments kit. AB - The first part of this study aimed to determine the effect of human variability on assessment using Semmes-Weinstein monofilaments. Fifteen healthy subjects were tested using a full monofilament kit of known calibration. When the subjects were retested, they responded to a different filament 48% of the time. The second part of the study aimed to determine the effects of vigorous hand activity on sensation, as measured with Semmes-Weinstein monofilaments. Forty randomly selected healthy subjects were assessed before and after they performed a brief, standardized gripping activity. After hand activity, the subjects responded to a different filament 53% of the time. In both parts of this study, the changes in monofilament results were random in direction and magnitude. The number of changes did vary between test sites, the radial nerve sites being most variable and the ulnar nerve sites least variable. It is concluded that the results of testing with the five smallest monofilaments in the full Semmes-Weinstein kit were not reliable in normal subjects and that they did not detect change in sensory function in normal subjects. PMID- 11866351 TI - An alternative splinting and rehabilitation protocol for metacarpophalangeal joint arthroplasty in patients with rheumatoid arthritis. AB - This prospective study was completed to establish results obtained using a static splinting regimen as an alternative to the dynamic extension splint. Fifteen patients with rheumatoid arthritis, who had undergone metacarpophalangeal (MCP) joint arthroplasties and a postoperative rehabilitation program of alternating MCP joint flexion and extension static splints, were assessed pre-operatively and reviewed postoperatively. Total active arc of MCP joint motion and ulnar deviation were measured, and an activities-of-daily-living questionnaire was completed by each patient at 19 months (mean) postoperatively. Mean total active arc of MCP joint motion showed statistical improvement from 21.6 degrees (range, 5 degrees-60 degrees) pre-operatively to 47.2 degrees (range, 15 degrees-84 degrees) postoperatively. The little finger gained the most improvement, with a 50.2 degree arc, showing that this regimen does not compromise flexion gains at this joint. Ulnar deviation improved from a mean of 30.4 degrees (range, 5 degrees-65 degrees) pre-operatively to 9.7 degrees (range, 0 degrees-30 degrees) postoperatively. These initial results reinforce the clinical impression that this alternating static splint regimen can be used as an effective alternative to the dynamic extension splint. PMID- 11866354 TI - A survey of the satisfaction of upper limb amputees with their prostheses, their lifestyles, and their abilities. AB - Seventy Australian upper limb amputees responded to a detailed postal questionnaire asking how often they wore their prostheses and their level of satisfaction with both their prostheses and their functional abilities. Fifty-six percent of amputees wore their limbs "once in a while" or "never." Prostheses were most often worn "all the time" for work and social activities. The amount of time amputees wore their prostheses was moderately associated with their level of satisfaction with their prostheses. The association between the amount of time amputees wore their prostheses and their level of satisfaction with their functional abilities was very low. Their prostheses were rated as "fair" or "not acceptable" by 64% of amputees. Sweating was rated as "not acceptable" by 55%. This may well be a significant contributing factor to the low prosthetic use. The amputees who did not wear prostheses did not have any greater satisfaction with their ability to do the tasks they want to do than the amputees who wore prostheses. PMID- 11866353 TI - How do therapists administer the rapid exchange grip test? A survey. AB - The rapid exchange grip (REG) test is one of many methods devised to detect insincere grip strength efforts. Studies investigating the REG test have used different testing protocols and different interpretation criteria for what constitutes a sincere effort. Since therapists get information about evaluation tools from the literature, inconsistencies among researchers may lead to a lack of standardization in the administration of the REG test among therapists. The purpose of the study was to survey therapists to determine what protocol they follow during REG test administration and how they interpret the REG test results. Fourteen therapists working in hand clinics completed the survey. The ways in which the therapists administered the REG test varied widely. Differences among therapists in administering the REG test included the position of the patient, the handling of the dynamometer, the number of repetitions, the hand switch rate, and the comparative test used to interpret the REG test. On the basis of these results, we concluded that the REG test does not meet the most basic testing and measurement criteria. This may impede the ability of therapists to report the outcomes of the REG test accurately and correctly. The implications for the profession due to the lack of standardization of the REG test are discussed. PMID- 11866355 TI - Evaluation of impaired fingertip texture discrimination and wrist position sense in patients affected by stroke: comparison of clinical and new quantitative measures. AB - Discriminative sensory loss is common following stroke but may not be adequately detected by routine clinical measures. Quantitative tests of texture discrimination and limb position sense have been recently developed. These tests provide reliable estimates of discrimination, differentiate impaired performance following stroke, and have standardized criteria of abnormality. The purpose of this study was to compare predictions of impairment based on current clinical measures with predictions based on these quantitative, standardized measures. Thirty-five patients who had strokes were tested on the new quantitative measures and clinical measures of texture discrimination and limb position sense. The findings indicated poor agreement between impairment defined using current clinical measures and statistically abnormal performance on the new quantitative, norm-referenced measures. The findings suggest that the quantitative tests may provide additional assessment information to supplement that of the existing clinical measures of texture discrimination and limb position sense. Further development of these new quantitative measures is indicated. PMID- 11866356 TI - A metacarpophalangeal joint stabilization splint: the Galindo-Lim thumb metacarpophalangeal joint stabilization splint. AB - Creation of a splint should allow for maximal functional use of the hand as well as comfort. The design of the Galindo-Lim splint enhances functional use of the hand not only because it stabilizes the MCP joint but also because it does not cross the wrist crease, thus allowing full wrist motion. Independence in the performance of daily activities is easily achieved with the Galindo-Lim splint (Figure 5). The splint is appropriate for patients with a variety of diagnoses, including MCP joint volar plate injuries, collateral ligament injuries, joint hypermobility, and arthritic conditions. PMID- 11866357 TI - A word about qualitative research. PMID- 11866358 TI - Hepatitis A vaccine: time for universal immunization. PMID- 11866359 TI - Fever: to treat or not to treat. Counterpoint. PMID- 11866360 TI - Fever: to treat or not to treat. Commentary. PMID- 11866361 TI - Self-concept in male and female adolescents with congenital heart disease. AB - Cognitive achievement, behavioural problems, and various dimensions of personality were assessed in 48 male and female patients with congenital heart disease (CHD) aged from 12 to 16 years in comparison to a control group. The CHD group showed a lower speed of cognitive processing but seemed to have less state anxiety and to possess a higher superego strength. Male adolescents with CHD presented with a reduced perceived capacity and self-esteem. This was not true for adolescent girls with CHD. The negative self-concept of boys with CHD may be partly explained by reduced physical ability interfering with peer relationships. PMID- 11866362 TI - Factors affecting the yield of pediatric EEGs in clinical practice. AB - Clinical factors affecting the yield of 2,500 pediatric electroencephalograms were analyzed. Electroencephalograms were interpreted as epileptiform in 40% of children with epilepsy. Most electroencephalograms were ordered for seizure in children not taking anti-epileptic drugs; just 15% showed epileptiform features. Six percent of electroencephalograms were epileptiform in non-seizure patients. The neurologist significantly influenced the odds of epileptiform interpretation (P = 0.022) and the recommendation to repeat the electroencephalogram (P < 0.001). In practice most electroencephalograms ordered for seizure are actually for non-seizure. In routine pediatric practice, electroencephalography has a low yield and appears to be over-used. PMID- 11866363 TI - Parental knowledge of and attitudes toward LYMErix (recombinant Osp--a lyme vaccine). PMID- 11866364 TI - Vocal cord dysfunction mimics asthma and may respond to heliox. AB - Vocal cord dysfunction (VCD), an under appreciated cause of wheezing, may be mistaken for or coexist with asthma. The vocal cords involuntarily adduct during inspiration, leading to inspiratory or biphasic wheezing. Asthma therapy offers no benefit and may result in injury. Proof of diagnosis requires endoscopy during an episode. Definitive therapy involves voice training by a speech pathologist, but heliox (20% to 40% oxygen in helium) has been used to reduce symptoms, resulting in dramatic improvement in wheezing and less anxiety. A retrospective review of recent experience with heliox treatment for patients with VCD was conducted, using a search of computerized inpatient and outpatient physician dictation reporting at Scott & White Memorial Hospital and Clinic. Five patients age 10 to 15 years were treated with a favorable response in four. There were no complications of therapy. A high index of suspicion can lead to the diagnosis of VCD, avoiding expensive, inappropriate, and harmful therapy. A trial of heliox inhalation for patients with symptomatic VCD may prove beneficial, analogous to the "reliever" role of beta agonists for asthma. Home or school use of heliox may reduce acute care visits, while voice training ("controller" therapy) is instituted. PMID- 11866365 TI - A young child with altered mental status. PMID- 11866366 TI - Prevention of serious bacterial infections in new-onset nephrotic syndrome: a survey of current practices. PMID- 11866367 TI - Propofol: risk vs. benefit. PMID- 11866368 TI - The use of haloperidol in agitated critically ill children. PMID- 11866369 TI - Reversal of obstructive sleep apnea in osteopetrosis following bone marrow transplantation. PMID- 11866370 TI - Bullous pemphigoid complicating Crohn's disease in a child. PMID- 11866371 TI - Osteonecrosis as a complication of treating acute lymphoblastic leukemia in children: a report from the Children's Cancer Group. PMID- 11866372 TI - Chest radiographs in the evaluation of febrile infants under 3 months of age. PMID- 11866373 TI - Achalasia in a child with tuberous sclerosis complex. PMID- 11866374 TI - Pediatric sledding injuries. PMID- 11866375 TI - Fever: to treat or not to treat. Point. PMID- 11866376 TI - Geriatric pathology a l'ancienne. AB - Long before modern medicine thought of old age as a disease, the Greeks and Romans appear to have referred to it as such and their learned men to have worked out theories on similar lines. Aristotle distinguished old age as a "natural" disease; both he and the Hippocratic school sought to devise a pathology of aging on the basis of the four "humours." Both were agreed that a loss of heat lay at the root of the matter, but Aristotle thought this was accompanied by dryness, the Hippocratic school by humidity. In the end it was the Aristotelian position which posterity accepted and embraced into the last century. PMID- 11866377 TI - Health concerns among India's elderly. AB - This article critically examines the health situation among elderly from two sets of independent sources. National Sample Survey in its 42 Round (July 1986-June 1987) canvassed a nationwide survey covering 50,000 households in 8312 villages and 4546 urban blocks in India to understand the socio-economic profile of aged persons. Secondly, an aging survey (1993) was conducted in Tamil Nadu, Kerala, Gujarat, and Karnataka states in India by the authors to study the elderly life in terms of general feeling, living arrangements, living support, social security, health, nutrition, their involvement in social and religious matters, views of old age homes and particularly life preparatory measures. The survey is based on interviews with about 2,253 persons aged 60 years and above, of whom 1325 are males and 928 are females. In fact, this survey is a part of a larger study conducted for the Social Development Section, Economic and Social Commission for the Asia and Pacific (ESCAP), United Nations, Bangkok, along with four other South East Asian countries-China, Korea, Singapore, and Thailand. From the survey conducted by us, we report only the health aspects of the elderly in this paper. The last section examines the major cause of death among elderly as reported from the Model Registration System of the Registrar General of India and concludes with feasible policy prescriptions. PMID- 11866378 TI - Comparing grandchildren's and grandparents' stake in their relationship. AB - This paper examines the intergenerational stake hypothesis in the grandparent grandchild relationship. The hypothesis predicts that older generations will perceive more closeness in a relationship than younger generations. Grandparent grandchild dyads are surveyed concerning various aspects of their relationship. Using measures of closeness and an established typology of grandparent-grandchild relationship style (Cherlin & Furstenberg, 1985), the study finds that grandparents perceive their relationship with their grandchildren to be considerably closer than their grandchildren perceive the relationship. However, their grandchildren perceive the relationship to be more active than their grandparents do. Explanations of these findings consistent with the intergenerational stake hypothesis are advanced. It is found that the two parties often disagree as to the nature of their relationship on the Cherlin and Furstenberg typology. Details of these disagreements are examined in detail, and implications for the grandparent-grandchild relationship are discussed. PMID- 11866379 TI - Meaning in life as an important component of functioning in old age. AB - The purpose was to locate different measures of meaning in life in a multidimensional space of functioning in old age. Fifty-five persons born in 1910 and 1914 participated in laboratory tests, a medical examination, and interviews over a three-year period from 1994 to 1997. Measures of meaning in life included a sense of meaning in life, a sense of coherence, and four indices derived from a life-line drawing (linearity, trend, mean level in the past, and mean level in the future). The other measures of functioning were the number of chronic diseases, functional disability, walking speed, self-rated health, cognitive capacity, provision of social relationships, loneliness, depressive mood, number of activities, life satisfaction, and wisdom. Using a multidimensional scaling method, a two-dimensional model of functioning was arrived at. In the first dimension, "subjective vs. objective," all the measures of meaning in life were located in the subjective half. The location of the measures along the second dimension, "psychosocial vs. physical," varied considerably. Some of the measures of meaning in life (a sense of coherence, life-line trend, and linearity) were located in the physical half, while others (sense of meaning in life, life-line mean level in the past) were in the psychosocial half The life-line mean level in the future was located in the center of the dimension. The space of functioning provided a novel and illustrative approach to functioning as a multifaceted behavioral competence with meaning in life as an integral part of it. PMID- 11866380 TI - Caregiver and elder experiences of Cambodian, Vietnamese, Soviet Jewish, and Ukrainian refugees. AB - Our purpose was to describe and compare Cambodian, Vietnamese, Soviet Jewish, and Ukrainian refugee caregivers and elders on life experiences, health status, and knowledge of available services. Detailed interviews were conducted with 105 female caregivers and 52 elders. Similar patterns emerged across all groups with regard to filial obligation, minimal knowledge of services, impact of immigration, and retention of cultural ties. Findings confirmed the special health and social service needs of refugee families in transition. PMID- 11866381 TI - High-performance liquid chromatographic analysis of free amino acids in fruit juices using derivatization with 9-fluorenylmethyl-chloroformate. AB - A simple, rapid, and reliable reversed-phase high-performance liquid chromatographic method for the analysis of 16 amino acids of main interest in commercial fruit juices (pear, orange, grapefruit, pineapple, peach, and apricot) is described. No sample cleanup is required. The pH of the fruit juices is adjusted to alkaline value (8.5) using 200 mM borate buffer, then amino acid is converted to stable derivatives using 9-fluorenylmethyl-chloroformate. The excess of derivatization reagent is removed by a hydrophobic amine, 1-amino-adamantane hydrochloride. The derivatization procedure is simple, fast, and described in detail. Amino acids are detected at 263 nm and eluted within 35 min. The calibration, precision (< or = 6.1%), and recovery (102% +/- 4%) of the method are reported. The conditions of separation are optimized; however, serine partially overlapped with aspartic acid. The amino acid profile of fruit juices is consistent with data from the literature. PMID- 11866382 TI - Automated headspace solid-phase microextraction and in-matrix derivatization for the determination of amphetamine-related drugs in human urine by gas chromatography-mass spectrometry. AB - An automated extraction and determination method for the gas chromatography (GC) mass spectrometry (MS) analysis of amphetamine-related drugs in human urine is developed using headspace solid-phase microextraction (SPME) and in-matrix derivatization. A urine sample (0.5 mL, potassium carbonate (5 M, 1.0 mL), sodium chloride (0.5 g), and ethylchloroformate (20 microL) are put in a sample vial. Amphetamine-related drugs are converted to ethylformate derivatives (carbamates) in the vial because amphetamine-related drugs in urine are quickly reacted with ethylchloroformate. An SPME fiber is then exposed at 80 degrees C for 15 min in the headspace of the vial. The extracted derivatives to the fiber are desorbed by exposing the fiber in the injection port of a GC-MS. The calibration curves show linearity in the range of 1.0 to 1000 ng/mL for methamphetamine, fenfluramine, and methylenedioxymethamphetamine; 2.0 to 1000 ng/mL for amphetamine and phentermine; 5.0 to 1000 ng/mL for methylenedioxyamphetamine; 10 to 1000 ng/mL for phenethylamine; and 50 to 1000 ng/mL for 4-bromo-2,5-dimethoxyphenethylamine in urine. No interferences are found, and the time for analysis is 30 min for one sample. Furthermore, this proposed method is applied to some clinical and medico legal cases by taking methamphetamine. Methamphetamine and its metabolite amphetamine are detected in the urine samples collected from the patients involved in the clinical cases. Methamphetamine, amphetamine, and phenethylamine are detected in the urine sample collected from the victim of a medico-legal case. PMID- 11866383 TI - A rapid and sensitive high-performance liquid chromatography assay for rofecoxib in human serum. AB - Rofecoxib is a selective cyclooxygenase (COX)-2 inhibitor that is approved for the treatment of acute pain and osteoarthritis in adults. A sensitive and rapid high-performance liquid chromatographic (HPLC) method of determining rofecoxib in human serum is described. Alkalinized plasma samples are extracted into an organic solvent containing an internal standard and evaporated under nitrogen. The dried sample residues are reconstituted with mobile phase and analyzed by HPLC. The method uses 100 microL of the sample and is linear from 20 to 2000 ng/mL of rofecoxib. Precision and accuracy studies are performed. Stability of the drug in serum over four weeks is documented. This new method is simple, sensitive, precise, and accurate. Its use will translate into faster laboratory turnaround time, and the small sample volume required (100 microL) makes this assay suitable for pediatric patients. This assay will expedite pharmacokinetic studies and the therapeutic drug monitoring of rofecoxib and possibly other COX-2 inhibitors. PMID- 11866384 TI - Simple determination of 22 organophosphorous pesticides in human blood using headspace solid-phase microextraction and gas chromatography with mass spectrometric detection. AB - A simple and rapid procedure for the determination of 22 organophosphorous pesticides (bromophos-ethyl, bromophos-methyl, chlorfenvinphos, chlorpyriphos, demethon-S-methylsulfon, diazinon, dichlorvos, dicrotophos, dimethoate, disulfoton, edifenphos, fenitrothion, fenthion, malathion, methidathion, mevinphos, monocrotophos, omethoate, parathion-ethyl, parathion-methyl, phosphamidon, and quinalphos) in human blood using headspace (HS) solid-phase microextraction (SPME) and gas chromatography (GC)-mass spectrometry (MS) is presented. The effects of various sample additions, incubation temperatures, absorption times, desorption times, and depths of fiber insertion into the injection port of the GC are optimized to enhance the sensitivity of the procedure. The recoveries of spiked blood samples are determined between 70% and 95% compared with samples prepared in water, and absolute recoveries are in the range between 0.1% and 19.6%. For quantitation in the single ion monitoring mode, linearity is established over concentration ranges from 0.025 to 5.0 microg/g with excellent coefficients of correlation (0.991-0.998). The detection limits are in the range between 0.01 and 0.3 microg/g. The time for analysis is 44 min per sample including extraction and GC-MS analysis. HS-SPME in combination with GC-MS is an effective method for the determination of organophosphorous pesticides in human blood and shows a great potential for use in rapid on-site analytical work, which is highly demanded in clinical and forensic toxicology. PMID- 11866386 TI - A rapid and sensitive high-performance liquid chromatography-electrospray ionization-triple quadrupole mass spectrometry method for the quantitation of oxycodone in human plasma. AB - A sensitive method for the determination of oxycodone concentrations in plasma by high-performance liquid chromatography (HPLC)-electrospray ionization-triple quadrupole mass spectrometry is described. The method is rugged, reliable, selective, and rapid with a run time of 2 min. One milliliter of plasma is made basic and extracted with 2-mL duplicate portions of 2% isoamyl alcohol in n-butyl chloride. The combined extracts are then evaporated to dryness, reconstituted in 100 microL of the mobile phase (15% methanol-85% water containing 0.1% acetic acid), and injected onto the HPLC. The limit of quantitation is 1 ng/mL, and the estimated limit of detection is 33 pg/mL (signal-to-noise = 3). Standard curves are linear over the range of 1 to 100 ng/mL with all correlation coefficient values greater than 0.9989. The method is used to determine the concentration of oxycodone in human plasma following the intravenous infusion of doses ranging from 5 to 15 mg in which the analysis of over 3000 plasma samples is required. PMID- 11866385 TI - Simultaneous and direct determination of oxalic acid, tartaric acid, malic acid, vitamin C, citric acid, and succinic acid in Fructus mume by reversed-phase high performance liquid chromatography. AB - A method for the simultaneous separation and direct determination of oxalic acid (OA), tartaric acid (TA), malic acid (MA), vitamin C (VC), citric acid (CA), and succinic acid (SA) in Fructus mume using reversed-phase high-performance liquid chromatography with a UV detector in an acidic medium is presented in this study. In the experiment, the optimization of chromatographic conditions (i.e., the pH and flow rate of the mobile phase, the absorption wavelength, and temperature of column) that affect the separation degree and peak shape of organic acids has been obtained. The linear ranges are found to be 0.05-4.7 microg for OA (r = 0.9999), 0.11-10.5 microg for TA (r = 0.9999), 0.114-11.4 microg for MA (r = 0.9999), 0.033-3.30 microg for VC (r = 0.9999), 0.155-15.5 microg for CA (r = 0.9998), and 0.194-19.4 microg for SA (r = 0.9996). For OA, TA, MA, VC, CA, and SA, the even recovery (n = 3) of six effective components are 100.9%, 99.97%, 101.2%, 102.1%, 101.1%, and 100.7%, respectively, and the largest relative standard deviation (n = 11) for the six components is less than 1.7%. The detection limits are 0.01 microg for OA, TA, and VC; 0.05 microg for MA; 0.03 microg for CA; and 0.1 microg for SA. In a single chromatographic run, OA, TA, MA, VC, CA, and SA can be determined in less than 7 min. The method can be used for the purpose of routine analysis and the quality control of a botanic (Fructus mume) containing these effective components. PMID- 11866387 TI - Analysis of amlodipine in human plasma by liquid chromatography-mass spectrometry. AB - A simple, sensitive, and specific liquid chromatographic method coupled with electrospray ionization (ESI)-mass spectrometry for the determination of amlodipine is developed. After extraction by ethyl acetate using nicardipine as the internal standard, solutes are separated on a C18 column with a mobile phase of methanol-1% HAc (65:35). Detection is performed on an air pressure ionization single quadruple mass spectrometer equipped with an ESI interface and operated in positive-ionization mode. Amlodipine quantitation is realized by computing the peak-area ratio (amlodipine-nicardipine) of the extracts analyzed in single ion monitoring mode (m/z 431 and m/z 480 for amlodipine and nicardipine, respectively) and comparing them with the calibration curve (r = 0.9991). PMID- 11866388 TI - What does HILIC mean? PMID- 11866389 TI - Why does the resolution decrease when carrier gas is changed from helium to hydrogen? PMID- 11866390 TI - Molecular modeling of quantitative structure retention relationship studies: retention behavior of polychlorinated dibenzofurans on gas chromatographic stationary phases of varying polarity by a novel molecular distance edge vector. AB - Chemical structures of polychlorinated dibenzodioxin (PCDD) congeners are described by a novel molecular distance edge vector (VMDE), developed in our laboratory, that consists of the modified molecular distance edge parameters based on the identical group as a pseudo-atom instead of a traditional atom. Quantitative structure retention relationships (QSRRs) between the new VMDE parameters and the gas chromatographic retention behavior of PCDDs are then generated by a multiple linear regression method for nonpolar, moderately polar, and polar stationary phases. All QSRR models with a high correlation (R > 0.99) are developed for nonpolar, moderately polar, and polar columns (DB-5, SP-2100, SE-54, and OV-1701). Cross validation with the leave-one-out procedure is performed, and satisfactory results are obtained with high correlation. The obtained results show that the new VMDE vector is adapted to characterize the chemical structure and model the retention behavior of PCDDs on various polar stationary phases. PMID- 11866391 TI - Proposed tenets of osteopathic medicine and principles for patient care. PMID- 11866392 TI - Managed care policy regarding OMT reimbursement illogical. PMID- 11866393 TI - Spirituality and end-of-life care. PMID- 11866394 TI - More on end-of-life care. PMID- 11866395 TI - Physician-patient interaction: what do elders want? AB - The purpose of this study was to identify communication styles and physician characteristics that correlate with improved patient adherence and satisfaction during geriatric healthcare interactions. A multiphase study design, incorporating the use of focus groups, socialization hours, educational seminars, and survey questionnaires, was used to discover the most effective methods for improving communication between physicians and their geriatric patients. Elders favored direct, interactive verbal communication over alternative communication styles such as role-playing activities or the use of visual aids. Chi-square analyses showed that men desired more time with medical providers than women, who instead expressed a preference for more thorough explanations of disease processes. Further, men--and African American men in particular--sought medical advice from trusted friends more frequently than did women, who often preferred to solicit medical advice from family members. The most significant barriers affecting physician-patient interaction were created by patients' inflated expectations for consultation time and by physicians' ineffective presentation styles. This study also reveals that physicians' characteristics and patients' gender and race also impacted the success of medical encounters. PMID- 11866396 TI - Obsessive-compulsive disorder: identification, neurobiology, and treatment. AB - Obsessive-compulsive disorder (OCD) is a common psychiatric disorder. It may go unrecognized, however, as many patients are embarrassed by their symptoms and are thus reluctant to report them. Recent research findings on OCD point to neurologic dysfunction in the circuitry of the orbitofrontal cortex and basal ganglia. The advent of the use of serotonin reuptake inhibitors (SRIs) as well as behavioral therapy has greatly improved treatment outcomes for patients with OCD. Given the likelihood that these patients are encountered in primary care consultations, physicians have the opportunity to play a crucial role in the early identification and proper treatment of OCD. PMID- 11866397 TI - Evaluation of spine injury in blunt trauma. AB - Although prehospital trauma guidelines call for spine immobilization for many trauma victims, there is a lack of clarity in medical institutions as to how trauma or emergency medicine physicians should proceed to remove cervical immobilization devices (CIDs) and "clear" the spine. Despite wide variations in physicians' approaches to such matters, however, certain specific aspects of vertebral assessment in such circumstances are well documented. The authors describe and explore several of these issues with respect to initial approach to the immobilized patient, clinical clearance of the spine, radiographic evaluation of the vertebrae in victims of blunt trauma, management of spine tenderness or pain, removal of CIDs, and indications for subspecialty consultation. Critical care physicians should be reminded that the responsibility lies with them for removing CIDs and halting other spine precautions--underlying the importance of careful consultation with radiologists and other specialists. PMID- 11866398 TI - Divalproex sodium in the treatment of migraine and cluster headaches. AB - The discovery of a new class of effective migraine-abortive medications, the triptans, has sparked a new interest in the study of vascular headache. Over the past few years, the Food and Drug Administration (FDA) has approved six new abortive pharmacologic therapies, with several others in various stages of clinical trials. Unfortunately, concurrent pharmacologic changes in headache prophylaxis have not kept pace with their abortive counterparts. However, divalproex sodium (Depakote), which is approved by the FDA as a migraine prophylactic agent, is the first in the anticonvulsant class of medication for migraine headache and has expanded the options in headache treatment. The objective of this retrospective multicenter study of 284 patients with migraine or cluster headaches was to examine the clinical efficacy and safety of divalproex sodium as prophylaxis in monotherapy and in polytherapy. Sixty-one percent of migraineurs and 73% of cluster patients noted a decrease in pain with divalproex sodium and continued that therapy for more than 3 months. Reported negative side effects included weight gain, nausea, somnolence, tremor, alopecia, dysequilibrium, and rash. However, only 14% of subjects discontinued therapy due to these side effects. Overall, divalproex sodium was found to be an effective and generally well-tolerated prophylactic treatment option as monotherapy or in polytherapy for migraine and cluster headache. PMID- 11866399 TI - Pain management in the culture of critical care. AB - Pain of critically ill patients is undertreated. Undertreatment of pain may be related, in part, to the culture of critical care practice, where nurses are challenged to meet competing patient demands. Implementation of appropriate pain management strategies is within the critical care nurse's scope of practice and must be a priority when delivering patient care. Although the multidisciplinary team can make the best holistic pain management plan, nurses have extensive independence and latitude in administration of pharmacologic and nonpharmacologic interventions. It is, in fact, "primarily the nurses' responsibility to administer the proper drug and dose at the proper time". Fostering a culture within critical care units that promotes optimal pain management is influenced by unit leadership, the values held by the staff, nurse competency, and an effective quality program that includes process and outcome indicators of pain management. PMID- 11866400 TI - Pathophysiology of pain in critically ill patients. PMID- 11866401 TI - Assessment of pain in the critically ill. AB - Accurate assessment of pain in the critically ill is undoubtedly a challenge. In this setting, however, the nurse can rise to the challenge and have a significant impact on pain management. Some careful thought and planning may allow the nurse to adapt usual assessment tools for use by patients who have difficulty in communicating. When patients cannot communicate, having accurate knowledge about pain and about the patient's condition can help nurses to make appropriate use of behavioral and physiologic signs. More research is necessary to test specific pain assessment tools in ICU settings. In addition, more studies are needed to document the reliability and validity of scales based on behavioral and physiologic indicators of pain in critical care settings. In the meantime, nurses can take several steps to ensure adequate pain assessment: (1) use all means possible to document the patient's self-report of the pain experience, (2) supplement these ratings with behavioral and physiologic indicators of pain status, and (3) document findings to comunicate the patient's pain to others caring for the patient. A careful thorough attempt to assess pain can ensure better pain, outcomes as well as a quicker and more positive return to health. PMID- 11866402 TI - Multimodal balanced analgesia in the critically ill. AB - Pain is common but is often undertreated in critically ill patients. A multimodal balanced analgesic approach is recommended for the management of pain in these patients. Balanced analgesia uses combined analgesic regimens, thereby reducing the likelihood of significant effects from a single agent or method. It may include several different drugs given to prevent or aggressively treat continuous and breakthrough pain as well as pain from procedures. PMID- 11866403 TI - Analgesia in the intensive care unit. Pharmacologic and pharmacokinetic considerations. AB - Morphine is the preferred analgesic agent for the critically ill patient. Fentanyl is the preferred analgesic agent for critically ill patients with hemodynamic instability or for patients manifesting symptoms of histamine release with morphine or morphine allergy. Hydromorphone is an acceptable alternative to morphine for patients with significant adverse affects from morphine or severe renal dysfunction. Meperidine and intravenous codeine should be avoided if at all possible. APAP and NSAIDs may be useful in the management of acute pain secondary to their opioid-sparing effects. Use of nonopioid analgesics may reduce the dose of opioid required for adequate pain control and help to minimize opioid-induced side effects. NSAIDs should be used only when the benefit-to-risk ratio is favorable. APAP should be used as an adjunct but not as the sole analgesic agent in critically ill patients. Regardless of which agent or agents are used to optimize pain control, it is imperative that caregivers recognize that the optimal analgesic dose and regimen vary widely between patients. Based on the pharmacokinetics and pharmacodynamics of the agent(s) selected, enough time for an adequate trial should be allowed before switching to other agents. Employing these principles optimizes the use of medications in the management of the complex physiologic response to pain. PMID- 11866404 TI - Nonpharmacologic treatment of pain. AB - Nonpharmacologic interventions for pain treatment are important complementary therapies but are not substitutes for pharmacologic management of pain. Use of nonpharmacologic pain treatments in critical care settings is helpful to decrease pain, but the challenge remains for nurses to have the knowledge, time, and skill to use these interventions in a busy daily practice with severely ill patients. Although numerous studies testing the effectiveness of nonpharmacologic interventions for pain management are available, the varying methods and interventions used in these studies make it difficult to draw conclusions. Further research on the use of these interventions for pain reduction is necessary to determine the most effective treatments and the conditions under which they should be used. PMID- 11866405 TI - Management of surgical and procedural pain in a critical care setting. AB - Critical care nurses can serve as pain management advocates so that those patients who undergo surgery and other procedures do not suffer needless pain. Research findings indicate that surgical and procedural pain can contribute to pathologic pain states related to nerve injury, tissue inflammation, and prolonged peripheral nociceptive input. Animal research findings support clinical practices that avoid the development of these conditions by effectively interrupting ongoing nociceptive input from the injured site. Knowledge of analgesic interventions, including pharmacologic and nonpharmacologic techniques, is essential to the professional practice of nursing the critically ill. The critical care nurse plays a pivotal role in preventing suffering, discomfort, and long-term negative consequences related to surgical and procedural pain. PMID- 11866406 TI - Managing the pain of traumatic injury. AB - Management of pain in the trauma patient is a complex issue requiring the ability to selectively match different injuries and patient situations with the most optimal pain management methods. Having an understanding of the various stages of trauma care helps clinicians to best support the goals of patient care while decreasing the detrimental effects of the stress response through good pain control interventions. When nurses have a good understanding of the various pain management interventions they are better able to assess the effectiveness, potential side effects, and goals of therapy. The following is a list of clinical pearls to help guide nurses to better manage the pain of traumatic injuries: Encourage your trauma team to standardize pain medications (particularly opioids). A protocol that uses a couple of opioids with varving routes of administration, onset, duration, mechanism of action, and side effects helps the team to become extremely familiar with them and better able to assess effectiveness and side effects. Frequent motor and sensory assessments are necessary in the injured-patient (especially with extremity and head injuries), and drug therapy choices must allow for a thorough baseline assessment and periodic checks to follow. Patients with multiple rib fractures or flail segments (particularly elderly patients) and no contraindications deserve serious consideration for treatment with an epidural. When using various pain management techniques, the nurse needs to be prepared to treat complications if they should arise. Airway equipment, drugs (i.e., oxygen, opioid antagonists, pressors), and resuscitation means must be immediately available. Nurses need to be extremely careful when receiving pain medication and other central nervous system depressant orders from various doctors involved in patients' care. If a pain management specialist is involved, all pain medication therapies should be supervised and ordered by that individual, particularly when spinal analgesia is employed. Nurses must be knowledgeable regarding the effects of spinal medications (local anesthetics and opioids) at varying spinal levels so as to assess therapeutic as well as untoward effects. Institute a diligent bowel protocol when using opioids. Opioid administration combined with the immobility and altered nutrition often associated with trauma can easily result in constipation, abdominal distention, and bowel obstruction. It is not uncommon for epidural blocks to need supplementation with other drug therapy, and this should not be considered a failure of the epidural. Any addition needs to be ordered and closely supervised by one primary team of doctors. Use of nonopioid drugs, if not contraindicated should be considered in all trauma patients. This is especially true for patients sustaining trauma and being discharged to home within 24 hours. They need to be educated about the pain they can expect the next day and how to judge if it is normal and expected or possibly the sign of a missed injury or developing complication (i.e., compartment svndrome infection). PMID- 11866407 TI - Assessment and management of acute cardiac chest pain. PMID- 11866408 TI - Managing cancer and noncancer chronic pain in critical care settings. Knowledge and skills every nurse needs to know. AB - Nurses and physicians caring for people with all types of pain are responsible for providing effective pain management in all clinical settings. Although the principles for managing all types of pain are similar, people with chronic pain have special needs to sustain optimal analgesia in critical care settings. Nursing practice must be guided by national standards, guidelines, and recommendations for managing chronic and acute pain. Pain assessment, reassessment, and follow-up; titration of medications to individual responses; aggressive management of side effects; prevention of pain; and routine evaluation of the effectiveness of the plan are basic skills for all health professionals. Every person's baseline level of pain must be determined, and preexisting interventions for pain relief must be maintained to assure continuity of care. Chronic pain complicates clinical problems and could have a profound effect on patient outcome. Managing chronic pain improves function, outcome, and quality of life. The knowledge, skills, medications, nondrug interventions, and technology are available to manage nearly all types of pain. Pain relief is a responsibility of all health care professionals. Our patients deserve our best efforts to optimize their comfort, and we must be accountable. PMID- 11866409 TI - Assessment and management of pain in the critically ill pediatric patient. AB - Alhough considerable progress has been made in the past decade in the study of pain in children, numerous myths, inadequate assessment, and insufficient knowledge of available treatment continue to contribute to the lack of effective management of children's pain. Clinicians who lack knowledge have misconceptions about pain contribute more to the problem than to the problem's solution. Adequate analgesia is to be provided to even the sickest child the medications, techniques, and approaches reviewed here. PMID- 11866410 TI - Pain management. An organizational perspective. AB - To sustain optimal pain-relieving care for acutely and critically ill patients, organizations must have systems in place to support evidence-based interventions, facilitate expert practice in the area of pain management, and encourage ongoing communication among patients, families, and providers. The key to success may be to create an organizational culture that is supportive of all practitioners to use effectively the many therapeutic options available to manage pain. Although redesigning structures to support pain management care can be done with relative ease, altering the culture of the practice environment and the behavior of care providers may prove more challenging. Cultural changes occur slowly, but as the practice changes do take hold, so do the results. The steps to change an organizational culture around pain management include understanding the existing system and involving key formal and informal leaders. Most experts recommend not attacking the existing culture head-on but living the culture you are trying to create and understanding that the transformation is not going to take place overnight. The use of evidence-based collaborative practice and quality improvement models may be a key to improving the practice environment for pain management. Evidenced-based pain management practice combined with an organizational commitment to optimal pain management for all patients helps to create and sustain the environment to effect the change. PMID- 11866411 TI - Monitoring and improving pain management practices. A quality improvement approach. AB - The development of a multidisciplinary pain management committee is a critical step in monitoring and improving pain management practices. The systematic evaluation of the quality of pain management using the principles of QI establishes a process that can lead to improvements in the care that patients receive. Clinicians need to develop a 1- to 2-year plan to improve the quality of pain management. Changes in clinicians' behaviors occur slowly; thus, members of pain management committees should not become discouraged but continue to move forward one step at a time. PMID- 11866412 TI - Moving to evidence-based practice for pain management in the critical care setting. AB - Widespread application of EBPM by bedside providers is needed to demonstrate the success of pain management strategies on patient outcomes. This goal is not easy to attain and generally requires time, patience, and a multidisciplinary team approach. Implementation and evaluation of pain interventions increase awareness and knowledge of pain management strategies and can result in an overall improvement in pain management. The literature and guidelines recommend the use of specific strategies to ensure practice change. Studies suggest that a more intensive or "active" effort to alter practice is generally most successful. The pain management program should be marketed so that both the health care providers and patients are aware of the goal and resources available. It may take 3 to 5 years to infuse the change and see the improvement. Reinfusion over time also needs to be planned. Bedside practitioners need to have knowledge of the current best evidence in pain management of the critically ill patient. Barriers to implementation must be eliminated so that practitioners can conscientiously and judiciously implement strategies to relieve pain. Opinion leaders and change agents need to be available to continually champion EBPM, and prompts to ask about pain should be provided to practitioners and patients. PMID- 11866413 TI - Hospital-acquired bacteraemia--surveillance and guidelines for practice. AB - In the Surveillance Report, bacteraemia was not defined specifically within the document. However, it was implied that bacteraemia was present if blood cultures were positive. No clinical information concerning the effect of bacteraemia on the patients or the degree of haemodynamic support required was described. Nor was the movement of patients between specialties indicated. This information would have been helpful to clinicians because it would demonstrate the increased severity of illness experienced by patients and the concomitant increase in services required to meet these needs, although of course prevention should be the key response to the data provided. It is evident from the two reports that infection of the bloodstream is described utilizing different terms, i.e. bacteraemia and catheter-related bloodstream infection. This is potentially confusing and efforts should be made to encourage the use of consistent terminology across specialties, e.g. infection control, critical care, oncology The potential for confusion also arises where evidence is pooled utilizing both CRI and CR-BSI as endpoints for the research reviewed. The recommendation associated with flushing the CVC with heparinized saline solution does not consider a patient's coagulopathy to be a contraindication, only the manufacturer's recommendations. This appears to be a limitation of the guidelines and may place the patient at risk. The recommendation also does not indicate if this is only to be performed when the catheter or lumen is not in use, or if heparin should be administered if it is in use. Finally, the categories used to denote the level of evidence are not defined in the document. It can be assumed that Category 1 relates to randomized controlled trials, which demonstrate homogeneity or/and narrow confidence intervals; whilst categories 2 and 3 relate to cohort studies and pooled data. It should be expected that a document of this nature should establish criteria or refer the reader to the primary source for categorization. Eggimann and Pittet (2000) have undertaken an excellent review of central venous catheter related infections in intensive care units. The evidence accrued and recommendations made mirrors the two reports outlined above. It is imperative we take action to reduce the incidence of CR-BSI and it is hoped that this Editorial has provided a basis for discussion and action; and will stimulate debate. PMID- 11866414 TI - Guidelines for nurse staffing in intensive care: a consultation document (3rd draft, July 2001). PMID- 11866415 TI - NHS modernisation agency--critical care programme. PMID- 11866417 TI - Working in collaboration with patients and relatives. PMID- 11866416 TI - The national service framework for coronary heart disease 'A blueprint for collaboration'. PMID- 11866418 TI - No going back: narratives by close relatives of the braindead patient. AB - The aim of this narrative study was to illuminate the meaning of being a relative of a patient diagnosed as brain dead. This has so far been explored only to a limited extent. By phenomenological-hermeneutic analysis of 14 narratives, a chronological narrative in 4 steps was identified: the disquieting event; the uncertain vigil; the arduous struggle; and the difficult road ahead. From the analysis, the metaphor of an inner journey emerged, starting from a life situation taken for granted and experienced as safe, and moving towards the unknown, the unfamiliar. During the journey, a series of events of decisive importance in the relative's life were found to take place. Major stages along the road were efforts made to comprehend the reality of death, saying farewell and taking leave of the loved one. The importance of the ICU nurse taking part in the relative's inner journey is discussed in relation to Eriksson's theory of suffering and Martinsen's theory of caring. Implications and suggestions for nursing care activities are discussed. PMID- 11866419 TI - The effects of music intervention on anxiety in the patient waiting for cardiac catheterization. AB - BACKGROUND: Hospitalization causes anxiety for many patients. It increases when patients anticipate their turn for cardiac catheterization. Music therapy reduces the psychophysiologic effects of anxiety and stress through the relaxation response. AIM: To determine the effects of music therapy an anxiety, heart rate and arterial blood pressure in patients waiting for their scheduled cardiac catheterization. METHODS: In a quasi-experimental, pretest-posttest design, 101 subjects were randomly assigned to either the test group: those who listened to 20 minutes of preselected music, or the control group: those who received treatment as usual. Subject anxiety levels and physiological values were measured while waiting their turn for cardiac catheterization and just prior to departure to the cardiac lab. RESULTS: 63 males and 38 females participated in the study. There was a statistically significant reduction in anxiety in the test group alone (P = 0.003) and in comparing the test to the control group (P = 0.004). In comparing the initial and departure physiologic values, it was noted that both heart rate and systolic blood pressure dropped in the test group, but increased in the control group. Within gender groups, there were no statistically significant differences in hemodynamics or STAI scores, but between gender groups there were significantly higher diastolic blood pressure in males and STAI initial and departure scores for females. DISCUSSION: Patients waiting for their cardiac catheterization benefit from music therapy. Anxiety and the heightened physiological values elicited by the stress response are reduced. Results also suggest that women waiting for cardiac catheterization experience a higher level of anxiety than males. PMID- 11866420 TI - Does extubation result in haemodynamic instability in patients following coronary artery bypass grafts? AB - Coronary heart disease and its management continue to be at the centre of Government health policy. The present political climate demands clinical effectiveness and best practice should be established, while maintaining the philosophy of cost-effectiveness and resource management. These directives have led practitioners to question the care of patients following coronary artery bypass surgery, in particular the role of mechanical ventilation and the subsequent act of extubation. A retrospective study of 89 patients who had coronary artery bypass grafts (emergency and elective) was undertaken, to establish if extubation had a significant effect on the haemodynamic status of patients with variable degrees of left ventricular function (19% with poor left ventricular function). The study found that extubation was achieved within a mean time of 4.97 hours following return from surgery. Extubation resulted in a significant increase in heart rate (P = 0.001), as well as a respiratory acidosis (pCO2: P = 0.000; pH: P = 0.000). However, the stability of the patient was not compromised, with neither mean arterial blood pressure (P = 0.825) nor oxygenation levels (P = 0.267) being significantly altered by extubation. On multivariate analysis, the act of extubation had no significant effect on any of the dependent variables. These results suggest that it is not extubation alone that has an impact on the haemodynamic stability of patients following coronary artery bypass grafts, but that this is indeed multifactorial. Therefore extubation is 'safe' practice for patients with varying degrees of left ventricular function following coronary artery bypass grafts. Limitations of the study are acknowledged. PMID- 11866421 TI - Perceptions and experiences of primary nursing in an ICU: a combined methods approach. AB - With the introduction of the Patient's Charter, greater emphasis has been placed on 'named nursing' (Department of Health 1991). While there is much literature extolling the benefits of this method of delivering care to patients (Reed 1988; Manley 1989; Macguire 1991); there is a dearth of empirical studies exploring primary nursing in an adult intensive care unit (ICU). In addition, little is known about how nurses feel about being a 'primary nurse'. The aim of this study was to determine qualified nurses' perceptions and experiences of the effect of primary nursing on patient care in an intensive care environment (ICU) and to explore nurses' experiences of being a primary nurse. This study was carried out in an ICU in Northern Ireland. Primary team nursing was the system of organizing care within this unit. This system consisted of teams of 'primary' and 'associate' nurses. A total of 65 qualified nurses were surveyed. Sixteen nurses took part in focus group interviews. A combined methods approach comprising a questionnaire and focus group interviews was employed for the study. Part one of the questionnaire provided data on professional and demographic details. Part two facilitated nurses to elaborate on their feelings and perceptions of the concept of primary nursing. The focus groups facilitated in-depth exploration of the respondents' personal feelings and experiences about their role as a primary nurse. The data obtained from the two-part questionnaire were analysed using descriptive statistic and content analysis. The data obtained from the focus groups were analysed using content analysis and the development of emerging themes. Analysis of the data revealed that the nurses' views were similar to those highlighted in the existing literature. Nurses perceived primary nursing to relate to the responsibility for the care of one patient, from admission to discharge with the primary nurse assessing, planning, implementing and evaluating care and the associate nurse assisting in the delivery of this care. Thus, continuity of care was seen as the major advantage of primary nursing. The findings, however, suggested that further teaching on the concept of primary nursing was required. In addition, many of the nurses admitted they experienced considerable stress, particularly in relation to their close proximity over a long period of time, with patients and their relatives. This is an important finding and highlights the need for primary nurses in ICU to have the opportunity (in some instances), to be relieved of their responsibility for a named patient, thereby reducing some of the potential for stress. It is acknowledged that the findings of this study may not be generalized beyond the identified sample. Further empirical studies exploring nurses' perceptions and experiences of primary nursing in an ICU are therefore needed. PMID- 11866422 TI - Crystalloid, colloid or small volume resuscitation? AB - Small volume resuscitation fluids are a combination of hypertonic crystalloid with a colloid. SVR fluids have the advantage of logistical convenience in the field situation. Clinical trials point to improved outcome in patients with penetrating trauma injury or traumatic brain injury. HSD may reduce the inflammatory process and has shown improved outcome in trauma patients who require intensive care. Trials so far have been carried out in small numbers of patients and improved survival benefit has only become evident on meta-analysis and sub-group analysis. Further large scale blinded randomized trials are required to confirm the promise of survival advantages. PMID- 11866423 TI - Chest X-ray quiz. A left-to-right shunt. PMID- 11866424 TI - Insights of high-density lipoprotein apolipoprotein-mediated lipid efflux from cells. AB - High-density lipoprotein (HDL) protects against cardiovascular diseases by removal of excess lipids from cells. HDL apolipoprotein-mediated lipid efflux involves multiple cellular proteins to remove both cholesterol and phospholipids that are otherwise stored in the cells. This article reviews recent progress in the understanding of receptors, signal mediators, Golgi and vesicle transport related to the pathway and proposes a model of HDL apolipoprotein receptor mediated exocytosis of cellular cholesterol. Such an exocytotic pathway could provide the most effective mechanism to remove excess cellular lipids and prevent atherogenesis. PMID- 11866425 TI - Gurken, a TGF-alpha-like protein involved in axis determination in Drosophila, directly binds to the EGF-receptor homolog Egfr. AB - The establishment of axial polarity in the Drosophila egg and embryo depends on intercellular communication between two cell types in the ovary, the germline, and the soma. The genes gurken and egfr encode two essential players of this communication pathway. Gurken protein, a TGF-alpha-like molecule, is expressed in the germline, while the EGF-receptor homolog, Egfr, is expressed in the somatic cells of the ovary. Using the yeast two-hybrid system we show here, for the first time, that Gurken protein directly binds to the extracellular domain of Egfr. This direct physical association requires the presence of an intact EGF motif within Gurken protein. Furthermore, we provide evidence that this characteristic motif may be sufficient for interaction with the receptor, at list in vitro. Our results firmly establish Gurken as the germline ligand of Drosophila Egfr. PMID- 11866427 TI - The CrkL adapter protein is required for type I interferon-dependent gene transcription and activation of the small G-protein Rap1. AB - We sought to determine the functional role of the CrkL adapter protein and downstream pathways in interferon signaling. In experiments using CrkL(--) mouse embryonic fibroblasts, we found that CrkL is required for IFN alpha-dependent gene transcription via GAS elements, apparently via the formation of DNA-binding complexes with Stat5. On the other hand, gene transcription via ISRE elements is intact in the absence of CrkL, indicating that the regulatory effects on gene transcription are mediated only via the formation of CrkL:Stat5 complexes. Our studies also indicate that activation of the small GTPase Rap1 by IFN alpha is defective in cells lacking CrkL, indicating that the protein plays a critical role in regulating activation of the growth inhibitory C3G/Rap1 pathway. The IFN alpha-inducible activation of the small GTPase Rap1 requires a functional N terminus SH3 domain in the CrkL protein, while the C-terminus SH3 domain does not appear to play a role in such a CrkL-function. We also demonstrate that both the Tyk-2 and Jak-1 kinases are required for activation of the CrkL/Rap1 pathway, as the Type I IFN-dependent GTP-bound form of Rap1 is inhibited by overexpression of dominant-negative Tyk-2 or Jak-1 mutants and is defective in cells lacking Tyk-2 or Jak-1. Taken altogether, these findings indicate that CrkL provides an important link between Jak-kinases and downstream cascades that play critical roles in IFN-dependent transcriptional regulation and induction of growth inhibitory responses. PMID- 11866426 TI - Mechanisms of the protective effect of L-alanine to D-galactosamine-induced hepatocellular injury: comparative studies of L-alanine and pyruvate. AB - The addition of L-alanine reduced lactate dehydrogenase leakage from primary cultured rat hepatocytes treated with galactosamine (D-gal), while D-alanine and other amino acids did not. However, the mechanisms have not yet been entirely clarified. In this study, we used various inhibitors of metabolism, i.e., aminooxyacetate, oligomycin, and quinolinic acid, to examine the relation between this protective effect and the metabolism of L-alanine. Quinolinic acid (10 mM) did not affect the hepatoprotective effect of L-alanine, while oligomycin (0.1 mug/ml) and aminooxyacetate (1 mM) eliminated the hepatoprotective effect of L alanine. L-Alanine also increased the albumin secretion by cultured hepatocytes treated with D-gal, while pyruvate had little effect. It was revealed that the intracellular content of pyruvate did not increase as a result of addition of L alanine. These results are consistent with the hypothesis that L-alanine metabolism is important for hepatoprotection, but pyruvate cannot be used as a substitute for L-alanine. PMID- 11866428 TI - Lysine can be replaced by histidine but not by arginine as the ER retrieval motif for type I membrane proteins. AB - The OST48 subunit of the oligosaccharyltransferase complex is a type I membrane protein containing three lysines in its cytosolic domain. The two lysines in positions 3 and 5 from the C-terminus are able to direct protein localisation within the endoplasmic reticulum (ER) by COPI-mediated retrieval. Substitution of these lysines by arginine resulted in cell-surface expression of OST48, whereas ER residency was maintained when either Lys-5 or Lys-3 but not both was replaced with arginine. Localisation of OST48 was not affected by substitution of the two lysines by histidine, indicating that a His-Xaa-His sequence, in contrast to Arg Xaa-Arg, contains ER-specific targeting information. These differences show that simple charge interactions are not sufficient for ER retention and that other structural factors also play a role. The His-Xaa-His sequence could represent a new and independent signal for directing ER localisation differing from both the arginine motif in type II proteins and the lysine motif in type I proteins. Our data do not exclude, however, that the histidine sequence simply mimics the lysine motif as a sorting signal, being recognised by and interacting with the same receptor subunit(s) in COP-I-coated vesicles. Conclusions arising from this assumption involving the conformation of lysine at the putative COP-I binding site and the failure of Arg-Xaa-Arg to mediate ER localisation for type I proteins are discussed. PMID- 11866429 TI - Detoxification of oxidized LDL by transferring its oxidation product(s) to lecithin:cholesterol acyltransferase. AB - In the present study, we isolated modified LCAT (m-LCAT) by hydroxyapatite column chromatography after incubation of crude LCAT (after DEAE SephadexA-50 column chromatography, penultimate step of LCAT purification) with oxidized LDL (oxLDL) at 37 degrees C for 1 h. The activity was found to be about 30% lower than that of native LCAT (n-LCAT). When activity was determined in the presence of oxLDL, m LCAT was less inhibited than n-LCAT by oxLDL. Treatments of purified LCAT either at 56 degrees C for 30 min, at 100 degrees C for 10 min, or with 6 mM 5-5' dithiobis-2-nitrobenzoic acid or 9 mM diisopropyl fluorophosphates (each at 37 degrees C for 30 min) resulted in the loss of its cholesterol-esterifying activity. When examined for their ability to detoxify oxLDL, native LCAT and LCAT treated at 56 degrees C for 30 min were found to detoxify oxLDL. These results indicate that oxidation product(s) of LDL is transferred and bound to LCAT in a way that does not depend on its cholesterol-esterifying activity, but rather on the availability of the sulfhydryl group of cysteine residue and the hydroxyl group of serine residue. PMID- 11866430 TI - Poly-L-lysine dissolves fibrillar aggregation of the Alzheimer beta-amyloid peptide in vitro. AB - beta-Amyloid peptide (beta A) is a major fibrillar component of neuritic plaques in Alzheimer's disease (AD) brains and is related to the pathogenesis of the disease. In this study, using electron microscopy, we describe herein the results concerning the efficacy of compounds that can dissolve preformed beta A fibrils in vitro. For such a purpose, two hydrosoluble and biocompatible polymers such as polyethylene glycol and poly-L-lysine were used. The poly-L-lysine appears as a potent dissolver of preformed beta A fibrils in vitro. Its efficiency is instantaneous. Poly-L-lysine can be used as a universal dissolver of all types of oligomeric beta-sheet conformation, precursor of the fibrils. This finding provides the basis for future investigation of the therapeutic potential of poly L-lysine in terms of preventing and/or retarding amyloidogenesis in AD and other types of amyloid-related disorders. PMID- 11866431 TI - Pyrococcus prefoldin stabilizes protein-folding intermediates and transfers them to chaperonins for correct folding. AB - A molecular chaperone prefoldin/GimC from the hyperthermophilic archaeum Pyrococcus horikoshii OT3 was characterized. Pyrococcus prefoldin protected porcine heart citrate synthase from thermal aggregation whereas each subunit alone afforded little protection. It also arrested the spontaneous refolding of acid-denatured green fluorescent protein and then transferred it not only to a group II chaperonin from the hyperthermophilic archaeum Thermococcus sp. strain KS-1, but also to a group I chaperonin from the thermophilic bacterium Thermus thermophilus HB8 for subsequent ATP dependent refolding. PMID- 11866433 TI - Cytotoxicity of IFN-gamma and TNF-alpha for vascular endothelial cell is mediated by nitric oxide. AB - Endothelial cell injury is a critical event in tissue damage accompanying inflammation, in which both inflammatory cytokines and reactive oxygen species may play pivotal roles, although the exact mechanism has not yet been clarified. We found that combined stimulation with interferon-gamma (IFN-gamma) and tumor necrosis factor-alpha (TNF-alpha) induced both cytotoxicity to murine vascular endothelial cell line F-2 and an increase in nitric oxide (NO). Therefore, in the present study, the implication of NO in cytotoxicity was examined. A potent iNOS specific inhibitor ONO-1714 completely blocked both cytokine-induced cytotoxicity and NO production. NO scavengers such as carboxy-PTIO and hemoglobin blocked cytotoxicity. Moreover, exogenous NO from NOC 18 also caused cytotoxicity. These results together demonstrated that cytotoxicity of IFN-gamma and TNF-alpha for endothelial cell F-2 was mediated by NO, suggesting a pathogenic role of cytokine induced NO production in endothelial damage under inflammatory conditions. PMID- 11866432 TI - Functional diversity among Notch1, Notch2, and Notch3 receptors. AB - To clarify functional diversities among the Notch receptors, we generated truncated forms of Notch1, Notch2, and Notch3 comprising the intracellular domain (aN1, aN2, and aN3) and investigated their transcriptional activities for HES1 and HES5 promoters driving the luciferase reporter gene (HES1-Luc and HES5-Luc). The reporter assays demonstrated that the transcriptional activities of aNs were markedly different from each other and dependent on the promoters examined. Furthermore, relative activities between some aN and another for each promoter were altered by the expression level of RBP-J kappa. We also found that the activities of aN1 and aN3 were reduced by coexpression of aN2. These observations suggest that each Notch receptor has a diverse role in the downstream gene expression and that the levels of HES1 and HES5 gene expression are complexly determined by various factors, such as the type and combination of the Notch receptors which confer the downstream signals and the expression level of RBP-J kappa. PMID- 11866434 TI - Global analysis of gene expression in renal ischemia-reperfusion in the mouse. AB - Ischemia-induced acute renal failure (ARF) is a relatively common disorder with major morbidity and mortality. To study global gene expression during ARF, 6-week old C57BL/6 male mice underwent 30 min of bilateral renal ischemia followed by reperfusion [I/R] or sham operation. Oligonucleotide microarrays [Affymetrix] with approximately 10,000 genes, 6,643 of which were present in mouse kidney, were used to analyze mRNA expression for up to 4 days following I/R. Fifty-two genes at day 1 and 40 at day 4 were up-regulated more than 4-fold [400%]. Seventy genes at day 1 and 30 genes at day 4 were down-regulated to under 0.25-fold from baseline [25%]. Real-time quantitative RT-PCR confirmed changes in expression for 8 genes of interest. Most of the induced transcripts are involved in cell structure, extracellular matrix, intracellular calcium binding, and cell division/differentiation. Our data identified several novel genes that may be important in renal repair after ischemia. PMID- 11866435 TI - Linear correlation between thermal stability and folding kinetics of lysozyme. AB - We have studied the refolding and thermal denaturation of hen egg white lysozyme in a wide range of pH values (from 1.5 to 9.4) using stopped-flow circular dichroism (CD) and differential scanning calorimetry (DSC). A linear correlation was found between the thermal denaturation temperature (T(m)) and the logarithm of the refolding rate of the slow folding phase of hen egg white lysozyme (lnk(2)). PMID- 11866436 TI - Molecular cloning and developmental expression of zinc finger transcription factor MTF-1 gene in zebrafish, Danio rerio. AB - Metal-responsive transcription factor, MTF-1 is a zinc finger protein, shown to be essential for embryonic development. Homozygous knockout mouse embryos for MTF 1 die in utero at day 14 of gestation, due to liver decay. In the present study, we report the complete nucleotide sequence of cDNA encoding zebrafish MTF-1 and the amino acid sequence similarity with that of mouse, human, fish and Drosophila. The size of the zebrafish MTF-1 cDNA is 3,379 bp and the coding region (1,779 bp) encodes a polypeptide of 593 amino acids. The putative zinc finger and transactivation domains comprised by zebrafish MTF-1 were also determined. The zebrafish MTF-1 shows high identity of 97, 93, 93 and 67% in the DNA binding zinc finger domain and 51, 44, 48 and 20% overall identity with fugu, human, mouse and Drosophila, respectively. RT-PCR results show the maternal expression of MTF-1 transcripts. The pattern of MTF-1 gene expression during embryonic and early larval development was studied by whole-mount in situ hybridization using DIG-labeled anti-sense RNA probe. Stronger and ubiquitous expression was observed during the embryonic stages whereas, specific expression, especially in the neural parts, was observed throughout the stages studied after hatching. PMID- 11866438 TI - The atomic structure of human methemalbumin at 1.9 A. AB - The high resolution structure of hemalbumin was determined by single crystal X ray diffraction to a resolution of 1.9 A. The structure revealed the protoporphyrin IX bound to a single site within a hydrophobic cavity in subdomain IB, one of the principal binding sites for long chain fatty acid. The iron is penta coordinated with the fifth ligand comprised of the hydroxyl oxygen of Tyr 161 (phenolic oxygen to heme plane distance: 2.73 A) in an otherwise completely hydrophobic pocket. The heme propionic acid residues form salt bridges with His 142 and Lys-190, which together with a series of hydrophobic interactions, enclose and secure the heme within the IB helical motif. A detailed discussion of the structure together with its implications for the development of potential blood substitutes is presented. PMID- 11866437 TI - Molecular basis for the inhibition of hypoxia-induced apoptosis by 2-deoxy-D ribose. AB - An angiogenic factor, platelet-derived endothelial cell growth factor/thymidine phosphorylase (PD-ECGF/TP), stimulates the chemotaxis of endothelial cells and confers resistance to apoptosis induced by hypoxia. 2-deoxy-D-ribose, a degradation product of thymidine generated by TP enzymatic activity partially prevented hypoxia-induced apoptosis. 2-Deoxy-D-ribose inhibits a number of components of the caspase-mediated hypoxia-induced apoptotic pathway. It inhibits hypoxia-induced caspase 3 activation, mitochondrial cytochrome c release, downregulation of Bcl-2 and Bcl-x(L), upregulation of hypoxia-inducible factor (HIF)-1 alpha, and loss of mitochondrial transmembrane potential in human leukemia HL-60 cell line. These findings suggest a molecular mechanism by which 2 deoxy-d-ribose confers the resistance to apoptosis. Thus 2-deoxy-D-ribose modulated suppression of HIF-1 alpha expression could prevent the hypoxia-induced decrease of the anti-apoptotic Bcl-2 and Bcl-x(L) on the mitochondria. 2-Deoxy-L ribose and its analogs may enhance apoptosis and suppress the growth of tumors by competitively inhibiting the activities of 2-deoxy-d-ribose and thus these analogs show promise for anti-tumor therapy. PMID- 11866439 TI - TEL Induces Aggregation in Transformed Cells and Induces Tube Formation in NIH3T3 UCLA Cells. AB - TEL/ETV6 is the frequent target of translocations associated with lymphoid and myeloid leukemias and solid tumors. We show that TEL induces aggregation of immortalized and transformed fibroblasts, endothelial cells and astrocytes. These aggregates form cellular cords in NIH3T3-UCLA by a cell autonomous process, which occurs when the monolayer is made up of over 75% of cells expressing exogenous TEL. Cords with a diameter of 15-25 microm contain a lumen and occur as tube structures. The possible relevance for vasculogenic mimicry is discussed. By contrast TEL did not induce aggregation of regular NIH3T3 cells, an effect that could only be induced by co-expression of oncogenic RAS/Lys12. Also transduction of TEL and RAS retroviral vectors into the endothelial MS1 cell line and TEL alone in the highly transformed glioblastoma cell lines EH-A and EH-B resulted in extensive aggregation. Thus, the induction of cellular aggregation by TEL correlates with transformation. PMID- 11866440 TI - Hyaluronan binding protein 1 (HABP1)/C1QBP/p32 is an endogenous substrate for MAP kinase and is translocated to the nucleus upon mitogenic stimulation. AB - The role of hyaluronan binding protein 1 (HABP1) in cell signaling was investigated and in vitro kinase assay demonstrated that it is a substrate for MAP kinase. Phosphorylation of endogenous HABP1 was also observed following treatment of J774 cells with PMA. HABP1 was coimmunoprecipitated with activated ERK, confirming their physical interaction in the cellular context. Upon PMA stimulation of normal rat fibroblast (F111) and transformed (HeLa) cells, the HABP1 level in the cytoplasm gradually decreased with a parallel increase in the nucleus. In HeLa cells, within 6 h of PMA treatment, HABP1 was completely translocated to the nucleus, which was prevented by PD98059, a selective inhibitor of ERK. We also observed that the nuclear translocation of HABP1 is concurrent with that of ERK, suggesting that ERK activation is a requirement for the translocation of HABP1. It is thus established for the first time that HABP1 is a substrate for ERK and an integral part of the MAP kinase cascade. PMID- 11866441 TI - Exip, a new alternative splicing variant of p38 alpha, can induce an earlier onset of apoptosis in HeLa cells. AB - One of the major families of the mitogen-activated kinases (MAPK), p38, has been shown to transduce extracellular stress stimuli into cellular responses. Among them, p38 alpha is the best characterized isoform and many biological phenomena, especially in the inflammatory responses, were attributed to the specific inhibitor-sensitive isoforms, namely p38 alpha and p38 beta. However, the roles played by each member are still unclear. Here, we report the identification of a new splice variant of p38 alpha, Exip (for exon skip), by RT-PCR using mRNA derived from a renal tumor cell line, OS-RC-2. Exip is predicted to encode a 307 amino-acid protein and the absence of exons 10, 11, and 11' results in the shift of the reading frame at the exon 9-12 junction to produce a unique 53-amino-acid C-terminus. The expression of mRNA was barely observed in cultured cells tested, but substantial amounts of mRNA were detected in mouse tissues. Unlike p38 alpha, Exip lost a common docking domain well conserved in major MAPK families for their specific interactions with upstream kinases or downstream substrates. Even though Exip is not phosphorylated at conserved TGY motif by p38-activating treatments, such as an osmotic shock or coexpression with a constitutive active form of MKK6 in HeLa cells, Exip can induce an earlier onset of apoptosis in HeLa cells. These results indicate that Exip has unique properties as a member of p38 alpha and may play role(s) in the signal transduction pathway(s) different from those of p38 alpha. PMID- 11866442 TI - Specific granules of human eosinophils have lysosomal characteristics: presence of lysosome-associated membrane proteins and acidification upon cellular activation. AB - Eosinophils possess characteristic specific granules. Their content may be important during host defense but it can also cause damage after release at sites of inflammation. We investigated possible lysosomal characteristics of these granules. Lysosome-associated membrane protein (LAMP)-1 and 2, were detected by Western blot, subcellular fractionation, and immunoelectron microscopy (IEM) and were localized to the membrane of specific granules and in vesicles of the cytoplasm, separate from secretory vesicles. No binding of mannose 6-phosphate receptor to proteins of specific granules could be detected, indicating that they are dephosphorylated and mature. Cellular activation by interleukin-5 caused acidification of specific granules, as detected by pH-dependent probes. The acidification was inhibited by concanamycin A (inhibitor of vacuolar H(+) ATPase). Activation of eosinophils by serum-treated zymosan (STZ) caused degranulation into STZ-containing phagosomes and incorporation of LAMPs to their membranes. In conclusion, specific granules of eosinophils can be regarded as specialized primary lysosomes, a feature that may be important for their function and integrity. PMID- 11866443 TI - Suicide gene therapy of human hepatoma and its peritonitis carcinomatosis by a vector of replicative-deficient herpes simplex virus. AB - Herpes simplex virus type 1 (HSV-1) deleted for the immediate-early gene was applied for treatment of hepatoma cells of SKHep 1 and Huh-7. Hepatoma cells were cultured in medium containing HSV1 expressing GFP gene (QOZ/HG) to determine its transfection rate, and both cell lines infected by MOI 1 of QOZ/HG were found to have high expression of GFP without cytotoxicity. Subcutaneous growth of SKHep 1 cell tumor in nude mice was significantly reduced by injection of replicative deficient herpes virus (TOZ.1) containing Tk-gene with administration of GCV, in comparison with that of noninjected tumor. SCID mice of peritonitis carcinomatosis due to Huh-7 hepatoma cells infected with TOZ.1 could survive longer under administration of GCV than those without TOZ.1. Therefore replicative-deficient HSV1 is a useful vector for treatment of human hepatoma cells, and TOZ.1 with GCV may be applied to suicide gene therapy for hepatoma and peritonitis carcinomatosis of hepatoma cells. PMID- 11866444 TI - Arsenic trioxide-induced apoptosis and its enhancement by buthionine sulfoximine in hepatocellular carcinoma cell lines. AB - We treated four hepatocellular carcinoma cell lines, HLE, HLF, HuH7, and HepG2 with ATO and demonstrated that arsenic trioxide (ATO) at low doses (1--3 muM) induced a concentration-dependent suppression of cell growth in HLE, HLF, and HuH7. HLE cells underwent apoptosis at 2 microM ATO, which was executed by the activation of caspase-3 through the mitochondrial pathway mediated by caspase-8 activation and Bid truncation. When these cell lines were exposed to ATO in combination with l-S,R-buthionine sulfoximine (BSO) which inhibits GSH synthesis, a synergistic growth suppression was induced, even in HepG2 showing a lower sensitivity to ATO than other cell lines tested. The intracellular GSH levels after the treatment with ATO plus BSO were considerably decreased in HLE cells compared with those after the treatment with ATO or BSO alone. The production of reactive oxygen species (ROS) which was examined by 2' ,7' dichlorodihydrofluorescein diacetate, increased significantly after the treatment with ATO plus BSO in HLE cells. These findings indicate that ATO at low concentrations induces growth inhibition and apoptosis, and furthermore that the ATO-BSO combination treatment enhances apoptosis through increased production of ROS in hepatocellular carcinoma cells. PMID- 11866445 TI - Helicobacter pylori activates the proto-oncogene c-fos through SRE transactivation. AB - Epidemiological studies have demonstrated a strong association between Helicobacter pylori infection and gastric cancer. However, there have been few detailed studies on the mechanism of cellular proliferation by H. pylori. Thus, we examined activation of the proto-oncogene c-fos to elucidate the underlying mechanism of cell proliferation caused by H. pylori. Activation of c-fos was evaluated in human gastric cancer cells (TMK1) by Northern blot and reporter assays with deletion analysis of the c-fos transcriptional control region. c-fos promoter activation and transcription were enhanced when cocultured with cag positive strains. H. pylori-mediated c-fos promoter activation was inhibited by MEK1/2 inhibitor (U0126). The deletion analysis indicated that serum response element (SRE) was required for the activation of c-fos by H. pylori. In conclusion, c-fos promoter activation and transcription were enhanced through the activation of extracellular signal-regulated kinases (ERK)/mitogen-activated protein kinase (MAPK) cascade in gastric cancer cells when cocultured with H. pylori possessing intact cag PAI. SRE is required for the activation of c-fos by H. pylori. These results suggest a direct involvement of H. pylori infection in cellular proliferation, which may play a role in neoplastic transformation. PMID- 11866446 TI - Biophysical characterization of cyclic nucleotide phosphodiesterases. AB - We have compared selected biophysical properties of three phosphodiesterases, from Arabidopsis thaliana, Saccharomyces cerevisiae, and Escherichia coli. All of them belong to a recently identified family of cyclic nucleotide phosphodiesterases. Experiments elucidating folding stability, protein fluorescence, oligomerization behavior, and the effects of substrates were conducted, revealing differences between the plant and the yeast protein. According to CD spectroscopy, the latter protein exhibits an (alpha + beta) fold rather than an (alpha/beta) fold as found with CPDase (A. thaliana). The redox dependent structural reorganization recently found for the plant protein by X-ray crystallography could not be detected by CD spectroscopy due to its only marginal effect on the total percentage of helical content. However, in the present study a redox-dependent effect was also observed for the yeast CPDase. The enzymatic activity of wild type CPDase (A. thaliana) as well as of four mutants were characterized by isothermal titration calorimetry and the results prove the requirement of all four residues of the previously identified tandem signature motif for the catalytic function. Within the comparison of the three proteins in this study, the PDase Homolog/RNA ligase (E. coli) shares more similarities with the plant than with the yeast protein. PMID- 11866447 TI - Identification and characterization of a novel type of membrane-associated prostaglandin E synthase. AB - Membrane-associated prostaglandin E synthase (mPGE synthase) was previously purified to apparent homogeneity from the microsomal fraction of bovine heart (Watanabe, K., et al., Biochim. Biophys. Acta 1439, 406--414, 1999). The N terminal 22-amino acid sequence of the purified enzyme was identical to that of the 88th to 109th amino acids deduced from the monkey (AB046026) or human (AK024100) cDNA that encodes a hypothetical protein with unknown function. The primary structure has the consensus region of glutaredoxin and of thioredoxin. We constructed an expression plasmid, using the vector (pTrc-HisA) and the monkey cDNA for the 290-amino-acid polypeptide. The recombinant protein with a M(r) of 33 kDa exhibited PGE synthase activity and was purified to apparent homogeneity by nickel-chelating column chromatography. The V(max) and K(m) values for PGH(2) of the purified recombinant mPGE synthase were about 3.3 mumol/min center dot mg of protein and 28 muM, respectively. The recombinant enzyme was activated by various SH-reducing reagents, i.e., dithiothreitol, glutathione (GSH), and beta mercaptoethanol, in order of decreasing effectiveness. Moreover, the mRNA distribution was high in the heart and brain, but the mRNA was not expressed in the seminal vesicles. These results indicate that the recombinant mPGE synthase is identical to the enzyme purified from the microsomal fraction of bovine heart, and is a novel type of mPGE synthase based on the primary structure, a broad specificity of thiol requirement, and tissue distribution. PMID- 11866448 TI - Inhibitory effects of IFN-gamma on HIV-1 replication in latently infected cells. AB - The progress in the use of HAART for the treatment of HIV-infected individuals has been limited by the development of viral resistance and the maintenance of viral latency. New therapeutic strategies geared toward improvement in the host's immune response are now being considered. We found that IFN-gamma induces CIITA through the JAK-STAT pathway and inhibits HIV-1 replication in latently infected cells. Its effect appears to be mediated through the reciprocal action of Tat and CIITA. With this beneficial effect, IFN-gamma and its inducers can be considered as an adjunct to the currently available therapy. We also addressed the safety of using simvastatin, an HMG-CoA reductase inhibitor, to treat dyslipidemia often associated with the use of protease inhibitors. Simvastatin did not show any unfavorable effects on HIV replication, thus could be used safely unless there are any drug interactions when administered. PMID- 11866449 TI - GC-rich elements flanking the transcription start site govern strong activation on the SNURF gene. AB - To study the regulation of the murine small nuclear RING finger protein SNURF (RNF4) gene, approximately 0.7 kb of its TATA-less promoter was isolated. This fragment conferred strong activation in reporter gene assays, yielding > or = 30% of the activity of the SV40 virus promoter/enhancer construct. Interestingly, the short region from -38 to +36 flanking the transcription start site was sufficient for potent basal promoter activity in various mammalian cell lines. Mutation of the conserved GC box at +9 abolished nuclear protein binding to the proximal promoter and severely compromised promoter activity, suggesting that this element is critical for the assembly of the transcription apparatus to regulate SNURF gene expression. Furthermore, our results show that the Wilms' tumor 1 gene product is one of the potential activators of the SNURF gene. PMID- 11866450 TI - Manipulation of unfolding-induced protein aggregation by peptides selected for aggregate-binding ability through phage display library screening. AB - A phage-displayed library of peptides (12-mer) was screened for the ability to bind to thermally aggregated bovine carbonic anhydrase (BCA), with a view toward examining whether peptides possessing this ability might bind to partially structured intermediates on the protein's unfolding pathway and, therefore, constitute useful tools for manipulation of the kinetic partitioning of molecules between the unfolded and aggregated states. Two peptides [N-HPSTMGLRTMHP-C and N TPSAWKTALVKA-C] were identified and tested. While neither showed thermal aggregation autonomously, both peptides individually elicited remarkable increases in the levels of thermal aggregation of BCA. A possible explanation is that both peptides bind to surfaces on molten BCA that are not directly involved in aggregation. Such binding could slow down interconversions between folded and unfolded states and stabilize aggregation-prone intermediate(s) to make them more prone to aggregation, while failing to achieve any steric prevention of aggregation. The approach has the potential of yielding useful aggregation aiding/inhibiting agents, and may provide clues to whether amorphous aggregates are "immobilized" forms of folding intermediates. PMID- 11866451 TI - Stabilization of vascular endothelial growth factor mRNA by hypoxia-inducible factor 1. AB - Hypoxia regulates expression of vascular endothelial growth factor (VEGF) by increasing its transcription and by stabilizing its mRNA. Despite the pivotal role of hypoxia-inducible factor 1 (HIF-1) in transcriptional activation of hypoxia-responsive genes, it is not known whether HIF-1 mediates hypoxia-induced stabilization of VEGF mRNA. We constructed adenoviral vectors expressing either the wild-type HIF-1 alpha (Ad2/HIF-1 alpha/FL), a constitutively stable hybrid form of HIF-1 alpha (Ad2/HIF-1 alpha/VP16), or no transgene (Ad2/CMVEV). In rat glioma (C6) cells and human cardiac, vascular smooth muscle, and endothelial cells, infection with Ad2/HIF-1 alpha/VP16 or Ad2/HIF-1 alpha/FL increased VEGF expression at both the mRNA and protein levels. Under normoxic conditions, the half-life of VEGF mRNA was 42 min in C6 cells. Hypoxia and Ad2/HIF-1 alpha/VP16 increased the half-life of VEGF mRNA to 3.3 and 2.7 h, respectively, while Ad2/CMVEV had no effect. These studies are the first to demonstrate that overexpression of HIF-1 alpha increases VEGF mRNA stability. Our results also suggest that stabilization of VEGF mRNA by hypoxia is mediated, at least in part, by HIF-1. PMID- 11866452 TI - Gonadotropin-releasing hormone stimulates annexin 5 messenger ribonucleic acid expression in the anterior pituitary cells. AB - We previously reported that annexin 5 is found specifically in gonadotropes and that the expression is dramatically enhanced after ovariectomy. In the present study, the expression of annexin 5 was examined in the primary culture of rat anterior pituitary cells using semiquantitative RT-PCR to determine if it is under the direct control of gonadotropin-releasing hormone (GnRH). Continuous administration of GnRH analog for 1 h enhanced the expression of both FSH beta subunit and annexin 5 mRNA. The expression of annexin 5 mRNA was also augmented by phorbol 12-myristate 13-acetate but not by forskolin. Administration of recombinant rat annexin 5 to the culture increased LH beta mRNA expression. These data clearly demonstrate that the expression of annexin 5 mRNA is directly controlled by GnRH and suggest that annexin 5 is involved in mediating GnRH action in the pituitary gland. PMID- 11866453 TI - Catalytic function of a novel protein protochlorophyllide oxidoreductase C of Arabidopsis thaliana. AB - In Arabidopsis thaliana Por C has been identified only on sequence homology to that of por A and por B. To demonstrate its catalytic function Arabidopsis thaliana protochlorophyllide oxidoreductase C gene (por c) that codes for the mature part of POR C protein having 335 amino acids was expressed in Escherchia coli cells. The POR C enzyme in the presence of NADPH and protochlorophyllide when incubated in dark formed a ternary complex. When it was excited at 433 nm, it had a fluorescence emission peak at 636 nm. After illumination with actinic cool white fluorescent light, a peak at 673 nm due to chlorophyllide gradually increased with concomitant decrease of 636 nm emission, demonstrating the gradual phototransformation of protochlorophyllide to chlorophyllide. The significance of differential por gene expression in light and dark among different species is discussed. PMID- 11866454 TI - Selection and stabilization of the RNA aptamers against the human immunodeficiency virus type-1 nucleocapsid protein. AB - The nucleocapsid (NC) protein of the human immunodeficiency virus-1 (HIV-1) plays an important role in the encapsidation of viral RNA and assembly of viral particle. Since the NC protein is resistant for mutation, it might be an excellent target for the anti-viral therapy. RNA aptamers that bind to the mature form of the NC protein were isolated from a RNA library. Surface plasmon resonance measurement and gel shift assay showed that the RNA aptamers specifically bind to the NC protein with high affinity and compete for the psi RNA binding to the NC protein. Mapping of the RNA aptamer showed at least two sites for the protein binding, suggesting a multiple and cooperative binding by the NC to RNA. In addition, the circular form of RNA avidly binds to the NC protein as a linear counter does. Stabilized RNA aptamer is expected to act as an inhibitor for the viral packaging. PMID- 11866455 TI - Mutational analysis of a mammalian reovirus mRNA capping enzyme. AB - The amino-terminal 42-kDa region of the 144-kDa mammalian reovirus lambda 2 protein is a guanylyltransferase. It catalyzes the transfer of GMP from GTP to the 5' end of 5' -diphosphorylated mRNA via a phosphoamide with Lys-190. This amino acid is located at the base of a deep cleft. Based on sequence comparisons, the Kx[V/L/I]S motif is present in all known and proposed guanylyltransferases of the family Reoviridae. The requirement for this conserved sequence and other regions of the enzyme was analyzed by site-directed mutagenesis. Based on the enzymatic activity of the mutants, Lys-190 and Asp-191 are the only amino acids of the (190)KDLS sequence that are necessary for enzymatic activity. Since Asp 191 has its side chain oriented away from the cleft, most likely it plays an indirect role in forming a functional guanylyltransferase. PMID- 11866456 TI - Cloning and expression of a lombricine kinase from an echiuroid worm: insights into structural correlates of substrate specificity. AB - Phosphagen kinases constitute a large family of enzymes catalyzing the reversible phosphorylation of guanidino acceptor compounds. These guanidino substrates differ substantially in size and chemical properties. In spite of the appearance of X-ray crystal structures for two members of this family, creatine kinase (CK) and arginine kinase (AK), the structural correlates of substrate specificity remain to be fully elucidated. We have determined the cDNA and deduced amino acid sequences for lombricine (guanidinethylphosphoserine) kinase (LK) from the echiuroid worm Urechis caupo and expressed the cDNA in Escherichia coli. The recombinant protein was purified by affinity chromatography and showed high capacity for phosphorylation of lombricine. Phosphagen kinases consist of a small, N-terminal domain and a much larger domain connected by a linker sequence. A key event in catalysis in CK and AK, and certainly all other phosphagen kinases, is a large conformational change involving involving a rotation of the two domains and the movement of two highly conserved flexible loops (one located in the small domain; the other located in the large domain of these enzymes) which clamp down on the substrates. Multiple sequence alignments of Urechis LK with the only other LK sequence available and CK, AK and glycocyamine kinase sequences, confirm the importance of the small flexible loop located in the N terminal domain of phosphagen kinases as one component of the structural determinants of guanidine specificity. The role of the other flexible loop in the large domain in terms of substrate specificity remains questionable. PMID- 11866457 TI - Overexpression of protein-tyrosine phosphatase PTP sigma is linked to impaired glucose-induced insulin secretion in hereditary diabetic Goto-Kakizaki rats. AB - The impaired glucose-induced insulin release in type 2 diabetes mellitus may be accounted for by reduced B-cell ATP/ADP ratio or decreased phosphorylation of proteins regulating exocytosis of insulin. This, in turn, could be due to enhanced phosphatase activity. Using in situ hybridization techniques to assess the expression of 11 different phosphotyrosine phosphatases (PTPases), known to be present in the B-cells, overexpression by approximately 60% of PTP sigma (also known as LAR-PTP2 or PTP NE-3) was demonstrated in pancreatic islets and liver of spontaneously type 2 diabetic Goto-Kakizaki (GK) rats. In agreement with these findings Western blot of islet lysates, using a polyclonal PTP sigma antiserum, showed increased amounts of the protein in GK relative to control rat islets. Exposure of isolated islets for 20 h to 5 muM antisense to PTP sigma, composed of an antisense PNA sequence of 15 bases linked to the cell penetrating peptide transportan, increased glucose-induced insulin secretion from GK rat islets, but not from control islets. In parallel, the amounts of the phosphatase decreased. In conclusion, increased expression of PTP sigma may be of pathogenetic significance for the defective insulin secretion in GK rat islets. PMID- 11866458 TI - Identification of AHNAK as a novel autoantigen in systemic lupus erythematosus. AB - To identify candidate autoantigens associated with arthritis, a rat chondrocyte cDNA library was immunoscreened with serum from a patient with rheumatoid arthritis. One isolated cDNA encoded part of AHNAK, a 700-kDa phosphoprotein with DNA binding properties, that appears to be involved in several signal transduction pathways. Immunoreactivity against an in vitro translated human AHNAK fragment was detected in 4.6% (5/109) of patients with rheumatoid arthritis, 29.5% (18/61) of patients with systemic lupus erythematosus (SLE), and 1.2% (2/172) of blood donors. Anti-AHNAK antibodies reacted with a recombinant human AHNAK fragment and with native AHNAK from C32 cell lysates. In vitro translated AHNAK fragment could be cleaved by granzyme B and caspase-3. Anti AHNAK positive SLE patients had a higher frequency of homogeneous antinuclear antibody staining patterns and a lower frequency of recent mucosal ulcerations. This is the first report that AHNAK can be targeted by the immune system in autoimmune disease. PMID- 11866459 TI - Antiperoxidase antibodies enhance refolding of horseradish peroxidase. AB - The effect of monoclonal antibodies on protein folding was studied using horseradish peroxidase refolding from guanidine hydrochloride as a model process. Among the five antiperoxidase clones tested, one was found to increase the yield of catalytically active peroxidase after guanidine treatment. The same clone also increased the activity of the native peroxidase by a factor of 2-2.5. While peroxidase refolding under standard conditions resulted in the recovery of only 7 8% of the initial catalytic activity, antibody-assisted refolding increased the yield to 50-100% (or 20-40% from the activity of native enzyme with antibodies). Kinetics of autorefolding and antibody-assisted refolding differed significantly. In the course of autorefolding the catalytic activity was recovered within the first 2.5 min and did not change further within a 2.5- to 60-min interval, whereas in the course of antibody-assisted refolding maximal catalytic activity was attained only in 60 min. The yield of active peroxidase for the antibody assisted refolding depended linearly on the antibody concentration. The observed effect was strongly specific. Other antiperoxidase clones tested as well as nonspecific antithyroglobulin antibody affected neither kinetics, no the yield of peroxidase refolding. PMID- 11866460 TI - A three-residue signal confers localization of a reporter protein in the inner nuclear membrane. AB - By alignment to the carboxy-terminal-deduced aa sequence of human cytomegalovirus glycoprotein B (gB), conserved hexameric aa motifs with putative function for localization in the inner nuclear membrane (INM) were identified in the nucleoplasmic tails of herpes simplex virus type 1 gB and of the cellular lamin B receptor. Fusion of the respective hexamers to CD8 as a reporter redirected transport of the chimeras into the INM, suggesting their functioning as modular signal motifs. Consecutive experiments showed that the three-residue motif RxR represents a consensus sequence which is sufficient for localization of the CD8 reporter in the INM. PMID- 11866461 TI - Constitutive TrkA activity in receptor-overexpressing PC12 clones. AB - We have studied ligand-independent signaling by the nerve growth factor receptor TrkA in PC12 clones, under conditions of receptor overexpression. Our results indicate that TrkA-overexpressing PC12 clones display constitutive receptor activation, involving both the mature, 140-kDa form and the immature, intracellular 110-kDa form of the receptor. Phosphorylation of Tyr 674/675, located in the activation loop domain and reflecting TrkA kinase activity, appears particularly prominent in the immature form of the receptor. Constitutive receptor activation is able to chronically stimulate the PI-3 kinase/Akt as well as the mitogen-activated protein kinase pathways, leading to ligand-independent neurite extension. Under conditions of overexpression, a significant fraction of the receptor is retained intracellularly by thiol-mediated mechanisms. Exposure of the cells to reducing agents promotes translocation of the intracellular pool of the receptor to the plasma membrane and suppresses ligand-independent neurite outgrowth. Our results suggest that the levels of expression of TrkA, both intracellularly and at the cell surface, may act to modulate its kinase activity and generate ligand-independent downstream signaling. PMID- 11866462 TI - Evidence for "pre-recruitment" as a new mechanism of transcription activation in Escherichia coli: the large excess of SoxS binding sites per cell relative to the number of SoxS molecules per cell. AB - In response to the oxidative stress imposed by redox-cycling compounds like paraquat, Escherichia coli induces the synthesis of SoxS, which then activates the transcription of approximately 100 genes. The DNA binding site for SoxS dependent transcription activation, the "soxbox," is highly degenerate, suggesting that the genome contains a large number of SoxS binding sites. To estimate the number of soxboxes in the cell, we searched the E. coli genome for SoxS binding sites using as query sequence the previously determined optimal SoxS binding sequence. We found approximately 12,500 sequences that match the optimal binding sequence under the conditions of our search; this agrees with our previous estimate, based on information theory, that a random sequence the size of the E. coli genome contains approximately 13,000 soxboxes. Thus, fast-growing cells with 4-6 genomes per cell have approximately 65,000 soxboxes. This large number of potential SoxS binding sites per cell raises the interesting question of how SoxS distinguishes between the functional soxboxes located within the promoters of target genes and the plethora of equivalent but nonfunctional binding sites scattered throughout the chromosome. To address this question, we treated cells with paraquat and used Western blot analysis to determine the kinetics of SoxS accumulation per cell; we also determined the kinetics of SoxS activated gene expression. The abundance of SoxS reached a maximum of 2,500 molecules per cell 20 min after induction and gradually declined to approximately 500 molecules per cell over the next 1.5 h. Given that activation of target gene expression began almost immediately and given the large disparity between the number of SoxS molecules per cell, 2,500, and the number of SoxS binding sites per cell, 65,000, we infer that SoxS is not likely to activate transcription by the usual "recruitment" pathway, as this mechanism would require a number of SoxS molecules similar to the number of soxboxes. Instead, we propose that SoxS first interacts in solution with RNA polymerase and then the binary complex scans the chromosome for promoters that contain a soxbox properly positioned and oriented for transcription activation. We name this new pathway "pre-recruitment." PMID- 11866463 TI - A novel interaction between thyroid hormones and 1,25(OH)(2)D(3) in osteoclast formation. AB - Thyroid hormones enhance osteoclast formation and their excess is an important cause of secondary osteoporosis. 3,5,3' -Triiodo-L-thyronine (T3) induced the mRNA expression of receptor activator of nuclear factor-kappa B ligand (RANKL), which is a key molecule in osteoclast formation, in primary osteoblastic cells (POB). This effect was amplified in the copresence of 1 alpha,25-dihydroxyvitamin D(3) (1,25(OH)(2)D(3)). Although T3 alone did not induce octeoclasts in coculture of bone marrow cells with POB, T3 enhanced 1,25(OH)(2)D(3)-induced osteoclast formation. Thyroxine (T4) also enhanced 1,25(OH)(2)D(3)-induced osteoclast formation. These data suggested that T4 was locally metabolized to T3 for its action, since T4 is a prohormone with little hormonal activity. The mRNA expression of type-2 iodothyronine deiodinase (D2), which is responsible for maintaining local T3 concentration, was induced by 1,25(OH)(2)D(3) dose- and time dependently. Our data would facilitate our understanding of the mechanism of osteoclast formation by thyroid hormones and suggest a novel interaction between thyroid hormones and 1,25(OH)(2)D(3). PMID- 11866464 TI - G beta gamma counteracts G alpha(q) signaling upon alpha(1)-adrenergic receptor stimulation. AB - In rat neonatal myocytes, a constitutively active G alpha(q) causes cellular injury and apoptosis. However, stimulation of the alpha(1)-adrenergic receptor, one of the G(q) protein-coupled receptors, with phenylephrine for 48 h causes little cellular injury and apoptosis. Expression of the G beta gamma-sequestering peptide beta ARK-ct increases the phenylephrine-induced cardiac injury, indicating that G beta gamma released from G(q) counteracts the G alpha(q) mediated cellular injury. Stimulation with phenylephrine activates extracellular signal-regulated kinase (ERK) and Akt, and activation is significantly blunted by beta ARK-ct. Inhibition of Akt by inhibitors of phosphatidylinositol 3-kinase increases the cellular injury induced by phenylephrine stimulation. In contrast to the inhibition of Akt, inhibition of ERK does not affect the phenylephrine induced cardiac injury. These results suggest that G beta gamma released from G(q) upon alpha(1)-adrenergic receptor stimulation activates ERK and Akt. However, activation of Akt but not ERK plays an important role in the protection against the G alpha(q)-induced cellular injury and apoptosis. PMID- 11866465 TI - Effect of resveratrol on growth of 4T1 breast cancer cells in vitro and in vivo. AB - In vitro, resveratrol inhibited growth of 4T1 breast cancer cells in a dose- and time-dependent manner. In vivo, however, resveratrol had no effect on time to tumor take, tumor growth, or metastasis when administered intraperitoneally daily (1, 3, or 5 mg/kg) for 23 days starting at the time of tumor inoculation. Resveratrol had no effect on body weight, organ histology, or estrous cycling of the tumor-bearing mice. Resveratrol, therefore, is a potent inhibitor of 4T1 breast cancer cells in vitro; is nontoxic to mice at 1-5 mg/kg; and has no growth inhibitory effect on 4T1 breast cancer in vivo. PMID- 11866466 TI - Antifungal mechanism of an antimicrobial peptide, HP (2--20), derived from N terminus of Helicobacter pylori ribosomal protein L1 against Candida albicans. AB - The antifungal activity and mechanism of HP (2-20), a peptide derived from the N terminus sequence of Helicobacter pylori Ribosomal Protein L1 were investigated. HP (2--20) displayed a strong antifungal activity against various fungi, and the antifungal activity was inhibited by Ca(2+) and Mg(2+) ions. In order to investigate the antifungal mechanism(s) of HP (2-20), fluorescence activated flow cytometry was performed. As determined by propidium iodide staining, Candida albicans treated with HP (2-20) showed a higher fluorescence intensity than untreated cells and was similar to melittin-treated cells. The effect on fungal cell membranes was examined by investigating the change in membrane dynamics of C. albicans using 1,6-diphenyl-1,3,5-hexatriene as a membrane probe and by testing the membrane disrupting activity using liposome (PC/PS; 3:1, w/w) and by treating protoplasts of C. albicans with the peptide. The action of peptide against fungal cell membrane was further examined by the potassium-release test, and HP (2-20) was able to increase the amount of K(+) released from the cells. The result suggests that HP (2-20) may exert its antifungal activity by disrupting the structure of cell membrane via pore formation or directly interacts with the lipid bilayers in a salt-dependent manner. PMID- 11866467 TI - YC-1-mediated vascular protection through inhibition of smooth muscle cell proliferation and platelet function. AB - YC-1, a synthetic benzyl indazole derivative, is capable of stimulating endogenous vessel wall cyclic guanosine monophosphate (cGMP) production and attenuating the remodeling response to experimental arterial angioplasty. In an effort to investigate the mechanisms of this YC-1-mediated vasoprotection, we examined the influence of soluble YC-1 or YC-1 incorporated in a polyethylene glycol (PEG) hydrogel on cultured rat vascular smooth muscle cell (SMC) cGMP synthesis, SMC proliferation, and platelet function. Results demonstrate that soluble YC-1 stimulated SMC cGMP production in a dose-dependent fashion, while both soluble and hydrogel-released YC-1 inhibited vascular SMC proliferation in a dose-dependent fashion without effects on cell viability. Platelet aggregation and adherence to collagen were both significantly inhibited in a dose-dependent fashion by soluble and hydrogel-released YC-1. Arterial neointima formation following experimental balloon injury was significantly attenuated by perivascular hydrogel-released YC-1. These results suggest that YC-1 is a potent, physiologically active agent with major anti-proliferative and anti-platelet properties that may provide protection against vascular injury through cGMP dependent mechanisms. PMID- 11866468 TI - A cellular mechanism that reversibly inactivates pancaspase inhibitor zAsp-CH(2) DCB: a potential pitfall causing discrepancy between in vitro and in vivo caspase assays. AB - Cell-permeable pancaspase inhibitors such as zAsp-CH2-DCB and zVAD-fmk are widely used to examine the involvement of caspases in cell death models. While examining the caspase-dependence of staurosporine (STS)-induced neuroblastoma cell death, we found that zVAD-fmk but not zAsp-CH2-DCB inhibits apoptosis. Time course analysis revealed that, in contrast to zVAD-fmk which constantly inhibited the processing of endogenous caspase substrates, zAsp-CH2-DCB inhibited substrate processing only for the first few hours after its addition to the culture medium. However, when the caspase activity in lysates prepared from cells treated with STS and zAsp-CH2-DCB was measured in vitro, quite unexpectedly, it was found that zAsp-CH2-DCB completely inhibits the STS-mediated activation of caspases throughout the observation period even when it apparently failed to inhibit the processing of caspase substrates within intact cells. These findings together suggest that there exists a cellular mechanism that inactivates zAsp-CH2-DCB in a reversible manner. This reversible inactivation was an active, intracellular process requiring de novo protein synthesis and was observed in another cell line HeLa and with different apoptotic stimuli such as ultraviolet irradiation. Our results have important implications that require consideration when designing experiments involving the use of caspase inhibitors as well as interpreting their results. PMID- 11866469 TI - Proteome analysis of hepatocellular carcinoma. AB - Development of hepatocellular carcinoma (HCC) is a complex process involving multiple changes in gene expression and usually occurs in the presence of liver cirrhosis. In this research, we observed proteome alterations of three tissue types isolated from livers of HCC patients: normal, cirrhotic, and tumorous tissue. Proteome alterations were observed using two-dimensional polyacrylamide gel electrophoresis and matrix-assisted laser desorption/ionization time-of flight mass spectrometry. Comparing the tissue types with each other, a significant change in expression level was found in 21 proteins. Of these proteins, sarcosine dehydrogenase, liver carboxylesterase, peptidyl-prolyl isomerase A, and lamin B1 are considered novel HCC marker candidates. In particular, lamin B1 may be considered as a marker for cirrhosis, because its expression level changes considerably in cirrhotic tissue compared with normal tissue. The proteins revealed in this experiment can be used in the future for studies pertaining to hepatocarcinogenesis, or as diagnostic markers and therapeutic targets for HCC. PMID- 11866470 TI - Down-regulated expression of exocytotic proteins in pancreatic islets of diabetic GK rats. AB - Exocytosis is regulated by exocytotic proteins, which are present in insulin secreting beta-cells and play regulatory roles in insulin secretion. Non-insulin dependent diabetes mellitus (type 2 diabetes) is a disease characterized by impaired insulin secretion and insulin resistance. Exocytotic protein immunoreactivities were studied in pancreatic islets of type 2 diabetic Goto Kakizaki (GK) rats using immunofluorescence histochemistry. The immunoreactivities for vesicle-associated membrane protein-2 (VAMP-2), synaptotagmin III, cysteine string protein (CSP), mammalian homologue of the unc 18 gene (Munc-18), alpha-soluble N-ethylmaleimide-sensitive attachment protein (alpha-SNAP), N-ethylmaleimide-sensitive factor (NSF) and synaptosomal-associated protein of 25 kDa (SNAP-25) exhibited weaker immunofluorescence intensity in islets of GK rats as compared to control Wistar rats. Insulin immunoreactivity was also decreased in GK rat beta-cells, whereas no detectable alterations in the expression of actin immunoreactivity could be detected. The data suggest that reduced expression of exocytotic proteins and decreased insulin content may contribute to the diabetic syndrome in the GK rat. PMID- 11866471 TI - Nuclear factor-kappa B-independent regulation of lipopolysaccharide-mediated interleukin-6 biosynthesis. AB - The possible involvement of nuclear factor (NF)-kappa B in mediating the regulation of interleukin (IL)-6 biosynthesis in response to E. coli-derived lipopolysaccharide-endotoxin (LPS) was investigated in vitro. In alveolar epithelial cells, irreversible inhibition of the proteasome complex by carbobenzoxy-L-leucyl-L-leucyl-L-leucinal (MG-132; 1-50 muM) did not affect LPS mediated IL-6 secretion. Whereas the selective inhibition of the NF-kappa B pathway by the action of caffeic acid phenyl ethyl ester (CAPE; 1-100 microM) attenuated LPS-dependent IL-6 production at 100 muM, sulfasalazine (SSA; 0.1--10 mM), a potent and irreversible inhibitor of NF-kappa B, did not inhibit LPS dependent IL-6 secretion. Incorporation of a selectively permeant inhibitor of NF kappa B, SN-50 (1-20 microM), a peptide which contains the nuclear localization sequence (NLS) for the p50 NF-kappa B subunit and the amino-terminal sequence of Kaposi fibroblast growth factor to promote cell permeability, did not reduce LPS mediated release of IL-6. These data indicate a NF-kappa B-independent pathway mediating LPS-dependent regulation of IL-6 biosynthesis in the airway epithelium. PMID- 11866472 TI - Biomarkers of human skin cells identified using DermArray DNA arrays and new bioinformatics methods. AB - Biomarker genes of human skin-derived cells were identified by new simple bioinformatic methods and DNA microarray analysis utilizing in vitro cultures of normal neonatal human epidermal keratinocytes, melanocytes, and dermal fibroblasts. A survey of 4405 human cDNAs was performed using DermArray DNA microarrays. Biomarkers were rank ordered by "likelihood ratio" algorithms and stringent selection criteria that have general applicability for analyzing a minimum of three RNA samples. Signature biomarker genes (up-regulated in one cell type) and anti-signature biomarker genes (down-regulated in one cell type) were determined for the three major skin cell types. Many of the signature genes are known biomarkers for these cell types. In addition, 17 signature genes were identified as ESTs, and 22 anti-signature biomarkers were discovered. Quantitative RT-PCR was used to verify nine signature biomarker genes. A total of 158 biomarkers of normal human skin cells were identified, many of which may be valuable in diagnostic applications and as molecular targets for drug discovery and therapeutic intervention. PMID- 11866473 TI - Macrosphelide B suppressed metastasis through inhibition of adhesion of sLe(x)/E selectin molecules. AB - Macrosphelide B (MSB), a 16-membered macrolide from Microsphaeropsis sp. FO-5050, inhibits adhesion of sialyl Lewis(x) (sLe(x))-expressing HL-60 cells to LPS activated (E-selectin-expressing) human umbilical vein endothelial cells (HUVECs) in vitro. This study examines MSB effects on metastasis of B16/BL6 mouse melanoma cells (B16/BL6 cells) and L5178Y-ML mouse lymphoma cells in vivo and analyzes the MSB antimetastatic activity mechanism. When administered MSB at 20 mg/kg/day, lung metastatic nodules of B16/BL6 cells were significantly decreased (T/C = 45%). However, no inhibition of metastasis of L5178Y-ML cells to the spleen and liver was observed. Flow cytometry analysis showed that B16/BL6 cells expressed high levels of sLe(x) antigen, whereas expression on L5178Y-ML cells was low. Under in vitro conditions, B16/BL6 cells demonstrated a greater degree of adhesion to LPS-activated HUVECs than L5178Y-ML cells, but adhesion was significantly inhibited by MSB and sLe(x) antibody. Combined therapy of MSB and cisplatin (CDDP) induced remarkable lung metastasis inhibition without adverse effects of CDDP to the host. All these findings suggest that MSB suppresses lung metastasis of B16/BL6 cells by inhibiting cell adhesion to endothelial cells through the sLe(x) molecule. PMID- 11866474 TI - NF-IL6 and HSF1 have mutually antagonistic effects on transcription in monocytic cells. AB - We have examined the functional antagonism between the regulator of the heat shock response, HSF1, and NF-IL6, which plays a major role in control of the acute phase response (APR). Agents that activate HSF1 such as heat shock and sodium salicylate inhibit NF-IL6 induced transcription while NF-IL6 activators such as lipopolysaccharide (LPS) and interleukin 6 (IL-6) repressed the stress responsive HSP70B promoter. In transfection studies, the inhibitory effects of HSF1 and NF-IL6 on the c-fms promoter were shown to be highly dose-dependent. Furthermore, heat shock is inhibitory to differentiation-linked expression of macrophage colony stimulating factor (M-CSF) receptor, product of the c-fms gene, which is transcriptionally activated by NF-IL6 but repressed by HSF1. Our studies suggest a strong mutual antagonism between the heat shock response and APR, which may influence the sensitivity and duration of inflammatory responses. PMID- 11866475 TI - Glucose-stimulated insulin secretion does not require activation of pyruvate dehydrogenase: impact of adenovirus-mediated overexpression of PDH kinase and PDH phosphate phosphatase in pancreatic islets. AB - Glucose-stimulated increases in mitochondrial metabolism are generally thought to be important for the activation of insulin secretion. Pyruvate dehydrogenase (PDH) is a key regulatory enzyme, believed to govern the rate of pyruvate entry into the citrate cycle. We show here that elevated glucose concentrations (16 or 30 vs 3 mM) cause an increase in PDH activity in both isolated rat islets, and in a clonal beta-cell line (MIN6). However, increases in PDH activity elicited with either dichloroacetate, or by adenoviral expression of the catalytic subunit of pyruvate dehydrogenase phosphatase, were without effect on glucose-induced increases in mitochondrial pyridine nucleotide levels, or cytosolic ATP concentration, in MIN6 cells, and insulin secretion from isolated rat islets. Similarly, the above parameters were unaffected by blockade of the glucose induced increase in PDH activity by adenovirus-mediated over-expression of PDH kinase (PDK). Thus, activation of the PDH complex plays an unexpectedly minor role in stimulating glucose metabolism and in triggering insulin release. PMID- 11866476 TI - Reduction of toxic metabolite formation of acetaminophen. AB - Acetaminophen is a widely used over-the-counter drug that causes severe hepatic damage upon overdose. Cytochrome P450-dependent oxidation of acetaminophen results in the formation of the toxic N-acetyl-p-benzoquinone-imine (NAPQI). Inhibition of cytochrome P450 enzymes responsible for NAPQI formation might be useful--besides N-acetylcysteine treatment--in managing acetaminophen overdose. Investigations were carried out using human liver microsomes to test whether selective inhibition of cytochrome P450s reduces NAPQI formation. Selective inhibition of CYP3A4 and CYP1A2 did not reduce, whereas the inhibition of CYP2A6 and CYP2E1 significantly decreased NAPQI formation. Furthermore, selective CYP2E1 inhibitors that are used in human therapy were tested for their inhibitory effect on NAPQI formation. 4-Methylpyrazole, disulfiram, and diethyl-dithiocarbamate were the most potent inhibitors with IC(50) values of 50 microM, 8 microM, and 33 microM, respectively. Although cimetidin is used in the therapy of acetaminophen overdose as an inhibitor of cytochrome P450, it is not able to reduce NAPQI formation. PMID- 11866477 TI - Modulation of sodium current in mammalian cells by an epilepsy-correlated beta 1 subunit mutation. AB - The syndrome of generalized epilepsy with febrile seizure plus (GEFS+) is associated with a single point mutation on the gene SCN1B that results in a substitution of the cysteine 121 with a tryptophane in the sodium channel beta 1 subunit protein. We have studied, in the HEK cells permanently transfected with the skeletal muscle sodium channel alpha-subunit (SkM1), the effects of a transient transfection of the wild type (WT) or C121W mutant beta 1-subunit. Coexpression of the WT beta 1 produces two effects on the sodium currents expressed in mammalian cells: the increase in the density of sodium channels, and the modulation of the inactivation of the sodium currents, inducing a hastening of the recovery from the inactivation. This modulation is less severe as observed when sodium channels are expressed in frog oocytes. We have observed that mutant C121W lacks this modulatory property, but maintains its property to increase the current density. Our observation suggests a possible involvement of this lack of modulation in the development of the GEFS+, providing the first hypothesis based on the observation of the functional properties of the beta 1-subunit C121W mutant in mammalian cells, which certainly represents a more physiological preparation, instead of in Xenopus oocytes, where the modulatory properties of the beta 1-subunit are artificially amplified. PMID- 11866478 TI - Peptide probe study of the critical regulatory domain of the cardiac ryanodine receptor. AB - The recently devised domain peptide probe technique was used to identify and characterize critical domains of the cardiac ryanodine receptor (RyR2). A synthetic peptide corresponding to the Gly(2460)-Pro(2495) domain of the RyR2, designated DPc10, enhanced the ryanodine binding activity and increased the sensitivity of the RyR2 to activating Ca(2+): the effects that resemble the typical phenotypes of cardiac diseases. A single Arg-to-Ser mutation made in DPc10, mimicking the recently reported Arg(2474)-to-Ser(2474) human mutation, abolished all of these effects that would have been produced by DPc10. On the basis of the principle of the domain peptide probe approach (see Model 1), these results indicate that the in vivo RyR2 domain corresponding to DPc10 plays a key role in the cardiac channel regulation and in the pathogenic mechanism. This domain peptide approach opens the new possibility in the studies of the regulatory and pathogenic mechanisms of the cardiac Ca(2+) channel. PMID- 11866480 TI - Ageing: definitions, mechanisms and the magnitude of the problem. AB - All multi-cellular organisms undergo change with time. Conception heralds the onset of growth and development, leading to reproductive competence and propagation of the species. With time, organisms age, leading to death as a final end-point. Whilst our knowledge and definitions of growth and reproduction are firmly established, the concept of ageing remains less well understood. One of the reasons for the lack of a singular definition of ageing is that it can be considered in many different ways, according to social, behavioural, physiological, morphological, cellular and molecular changes. Research has led to a number of theories being proposed that may explain the ageing process. In this chapter, we will review some of these theories and address some of the following fundamental questions: What is ageing? How can ageing be measured? When does ageing begin? When is an organism defined as old? PMID- 11866479 TI - Dilinoleoylphosphatidylcholine is responsible for the beneficial effects of polyenylphosphatidylcholine on ethanol-induced mitochondrial injury in rats. AB - Chronic ethanol consumption depletes phosphatidylcholines (PC) in membranes and hepatic mitochondria are an early target of this toxicity. Our previous studies showed that soybean-derived polyenylphosphatidylcholine (PPC), attenuated mitochondrial liver injury. Since dilinoleoylphosphatidylcholine (DLPC) is the major component of PPC, we assessed whether it is responsible for the protection of PPC. Forty-two male rats were fed the following liquid diets for 8 weeks: Control; Control with DLPC (1.5 g/1000 Calories (Cal); Alcohol (36% of Cal); Alcohol with DLPC (1.5 g/1000 Cal) and Alcohol with PPC (3 g/1000 Cal). As expected, ethanol feeding diminished the capacity of hepatic mitochondria to oxidize glutamate and palmitoyl-1-carnitine, and also decreased the activity of mitochondrial cytochrome oxidase. These effects were equally prevented by either PPC or DLPC. In conclusion, DLPC fully reproduced PPC's protective action and may be effective in the prevention or delay of more severe liver damage. PMID- 11866481 TI - Assessing and interviewing the elderly: interpretation of signs and symptoms. AB - Interviewing the elderly patient, typified by poor memory, often confused and sometimes hard of hearing, requires great patience and perseverance on the part of the physician to extract pertinent information from a complicated history involving past and present illnesses, multiple medications (both prescribed and over-the-counter), and social as well as economic issues. These circumstances may include retirement, death of a spouse and a change of living conditions. Assessment of these issues, followed by a careful physical examination, must lead to a diagnostic programme that is thorough yet practical, with consideration of the benefit of each procedure contemplated. The ultimate goal must be to renew the patient's ability to function as well as to improve the patient's quality of life. Many illnesses characteristic of the aged person are treatable but not curable. The goal is to improve the quality of life and to make the declining years as comfortable as possible. Typical cases illustrating these points are presented and discussed and their resolutions described. PMID- 11866482 TI - Nutrition in the elderly. AB - Malnutrition is more common in elderly persons than in younger adults. Ageing itself, however, neither leads to malabsorption nor to malnutrition with the exception of a higher frequency of atrophic gastritis in older persons. Malnutrition in elderly people is therefore a consequence of somatic, psychic or social problems. Typical causes are chewing or swallowing disorders, cardiac insufficiency, depression, social deprivation and loneliness. Undernutrition is associated with a worse prognosis and is an independent risk factor for morbidity and mortality. Awareness of this problem is therefore important. For the evaluation of nutritional status, it must be remembered that most normal values are derived from younger adults and may not necessarily be suitable for elderly persons. Suitable tools for evaluating the nutritional status of elderly persons are e.g. the body mass index, weight loss within the last 6 months, the Mini Nutritional Assessment (MNA) or the Subjective Global Assessment (SGA). An improvement in the nutritional status can be achieved by simple methods such as the preparation of an adequate diet, hand feeding, additional sip feeding or enteral nutrition. PMID- 11866483 TI - Nutrition and oral health. AB - Reduced chewing function in community-dwelling older people with adequate general health is linked to having fewer than 20 teeth present or to wearing removable dentures. By chewing for longer periods of time or swallowing larger food particles they are normally able to compensate for the impaired function. The masticatory function can be restored by adequate prosthetic therapy, which results in increased activity of the masticatory muscles during chewing and reduces the chewing time and the number of chewing strokes until swallowing. In frail or dependent elderly people undernutrition is prevalent because of health problems, reduced appetite and poor quality of life. Poor oral health and xerostomia are often associated with a reduced body mass index and serum albumin level and the avoidance of difficult-to-chew foods. Maintenance or re establishment of masticatory function is an integral part of the medical health care of these patients, with the aim of improving their nutritional status and quality of life. PMID- 11866484 TI - Drug metabolism and drug interactions in the elderly. AB - In the elderly concomitant use of several drugs (polypharmacy) is very common. Thus, the risk for drug interactions might be increased in this population. Since most drugs are hepatically eliminated by various metabolic pathways, liver function has to be considered as an additional factor modifying drug response. This chapter focuses on the hepatic mechanisms of interactions, especially on various inhibitors and inducers of the most important cytochrome P450 isoenzymes involved in drug metabolism. In addition, age-dependent changes in liver function are addressed. Based on pharmacokinetic results with different probe drugs, some inconsistencies in this area are discussed. The most important metabolic drug drug interactions are independent of the age of the patients. However, since elderly patients consume a greater proportional share of drugs, they represent a population at risk for interactions. Awareness of this clinical problem may help to diminish those risks. PMID- 11866485 TI - Physiology and pathology of the oesophagus in the elderly patient. AB - In the 1960s, the term 'presbyoesophagus' was introduced for what were felt to be the characteristics of the oesophagus in old age. Since then a number of sophisticated studies using modern manometric, radiological, pH-metric or endoscopic equipment have been performed to better study this subject. Although results in some aspects are still contradictory, these studies have provided us with some more detailed information on the physiology of the ageing oesophagus. Beginning with an overview of what is currently known and discussed about age induced physiological changes in oesophageal function, this review will then focus on specific problems of oesophagus-related diseases in the elderly. The main topics discussed will include presentation, diagnosis and treatment of primary and secondary motility disorders, oesophageal manifestations of neuromuscular and neurological disease, gastro-oesophageal reflux disease and oesophageal cancer in the elderly. PMID- 11866486 TI - Physiology and organ-related pathology of the elderly: stomach ulcers. AB - Peptic ulcer disease, particularly as a result of its complications, is a burden that is focused on the elderly through their higher Helicobacter pylori prevalence and use of non-steroidal anti-inflammatory drugs (NSAIDs). In these patients, senescence may further increase ulcer susceptibility, particularly in the stomach, by the loss of mucosal protection and repair mechanisms. Age is mainly a marker for the increased prevalence of other complicated ulcer risk factors such as previous ulcer history and use of anti-coagulants, steroids and aspirin. The development of selective cyclo-oxygenase inhibitors (coxibs) has reduced the specific risk of NSAID ulceration, but the residual incidence in high risk patients remains substantially higher than that in young patients without other risk factors. The argument for early surgery versus endoscopic therapy in high risk patients with bleeding ulcers has not been resolved, both having a high mortality. There is still potential for the development of new strategies to prevent primary and secondary ulcers, either by new drug development or by expanding existing co-prescription strategies. PMID- 11866487 TI - Gastrointestinal bleeding in the elderly. AB - Gastrointestinal bleeding in elderly individuals is a frequent cause of consultation with a physician and of hospital admissions. Co-morbidity and greater medication use in this steadily growing patient group influence the clinical course and adversely affect outcome. Clinical presentation is often predictable and guides subsequent patient management. Due to a surprising lack of prospective controlled data in the area of gastrointestinal bleeding, the selection of diagnostic and therapeutic manoeuvres often depends more on local expertise and availability than on an algorithmic approach. Advances in endoscopic, medical, radiological and surgical treatment modalities offer promising new diagnostic and therapeutic tools, particularly in concerted applications. Outcome studies on the appropriate sequence and linking of these modalities are urgently needed. This chapter will address clinical presentation, aetiology, diagnosis and treatment of both upper and lower gastrointestinal bleeding in the elderly. PMID- 11866488 TI - Atrophy-metaplasia-dysplasia-carcinoma sequence in the stomach: a reality or merely an hypothesis? AB - The results of recent investigations have suggested that the old hypothesis of an atrophy-metaplasia-dysplasia-carcinoma sequence in the stomach needs to be qualified. The most common cause of intestinal metaplasia is Helicobacter pylori gastritis. The consequence of this intestinal metaplasia is focal atrophy. Helicobacter pylori infection may also trigger an autoimmune gastritis of the corpus mucosa, with atrophy and intestinal metaplasia. Most intestinal metaplasias are only 'paracancerous' but not 'precancerous' lesions. Diffuse gastric carcinomas, such as the signet ring cell carcinoma, arise independently of intestinal metaplasia. Histogenetically, numerous carcinomas of the stomach are primarily of the gastric type, and may secondarily change into the intestinal type.High-grade intra-epithelial neoplasias (dysplasias) detected during the biopsy-based diagnostic work-up appear to be a marker for carcinoma and must, therefore, be removed endoscopically. The detection of intestinal metaplasia in routinely obtained biopsy material is subject to sampling error and is, therefore, not a suitable marker for an increased risk of a gastric carcinoma developing. As an alternative, the concept of gastritis of the carcinoma phenotype, which is more frequently found in early gastric carcinomas and in the relatives of gastric carcinoma patients, has been developed. In this concept, the diffuse parameters of grade and activity of the gastritis in the antrum and corpus, which are independent of sampling error, are subjected to a comparative analysis. A risk gastritis of the carcinoma phenotype is diagnosed when the grade and activity of the gastritis in the corpus are at least equally as pronounced as in the antrum. Currently, this concept is being tested in a prospective ongoing study. Future studies must show whether, and if so which, immunohistochemical or molecular-genetically detectable changes can be applied as risk markers in the diagnostic work-up. Helicobacter pylori eradication probably does not lead to complete regression of the intestinal metaplasia and ensuing focal atrophy. However, eradication of H. pylori does lead to the normalization of changes that can lead to mutations of the stem cells of the gastric mucosa (free radicals, nitric oxide, cell proliferation and vitamin C secretion). PMID- 11866489 TI - Endoscopy in the elderly patient. AB - In the elderly who require endoscopy for diagnosis and /or intervention, the endoscopist should be aware of the special risks related to the presence of concomitant systemic diseases. This is especially pertinent in the use of sedatives and analgesics due to the altered physiological functions related to ageing. This can be further complicated by the fact that elderly patients are often prescribed multiple drugs, which makes for the possibility of serious drug interactions. Endoscopy is a minimally invasive technique that is safer than conventional surgery in many conditions. The endoscopist must take into consideration the important factors related to quality of life, as well as the wishes of the patients and their families. The endoscopist must be sympathetic to their wishes and realize when investigation and treatment are not appropriate. PMID- 11866490 TI - Care of the elderly with gastrointestinal problems in family practice. AB - Gastrointestinal problems in older people cause a great amount of anxiety, morbidity and mortality. In general these diseases present for the first time to family practitioners. The management of gastrointestinal problems is more difficult because in an older age group, functional diseases can present in the same way as organic diseases. In addition, family practitioners see a different kind of patient than speciality physicians and may not have immediate access to diagnostic investigations. In this chapter, the role of the family practitioner in screening for gastrointestinal problems in asymptomatic older people is explored. In addition, how they differentiate between organic and non-organic disease and refer appropriately to secondary care is discussed. The role of family practitioners in the on-going maintenance of gastrointestinal diseases and in the management of Helicobacter pylori in community dwelling older people is also considered. PMID- 11866493 TI - Forceps delivery. AB - This chapter reviews the role for instrumental delivery in current practice and gives a critical account of current techniques of operative vaginal delivery by forceps. Important issues, including case choice, documentation, forceps application, avoidance of complications, and training and education are included. PMID- 11866494 TI - Vacuum-assisted delivery. AB - Unsuccessful vacuum extraction, cup detachment and failed anterior rotation in occipitoposterior positions are commonly associated with obstetric factors that are avoidable or correctable. These factors include the preferential use of soft vacuum cups, incorrect cup applications and attempts to deliver with the vacuum extractor before the cervix is completely dilated. Evidence from randomized trials demonstrates that soft cups cause fewer cosmetic effects and scalp lacerations than rigid cups. Soft cups do not reduce the incidence of cephalhaematomas nor have they been shown to provide any advantage over rigid cups for the prevention of subgaleal haemorrhage. Clinically significant subgaleal haemorrhage and intracranial injury are almost always preceded by difficult vacuum extraction. Although the vacuum extractor is less likely than forceps to injure the mother's genital tract and anal sphincters at delivery, no significant differences have been demonstrated between the instruments in terms of subsequent urinary or bowel disturbances. PMID- 11866495 TI - Breech delivery. AB - Fetuses that present by the breech are at increased risk of trauma and hypoxia during delivery. The threshold for Caesarean section for breech presentation had been low for several years. The result of the term breech trial confirms that planned Caesarean section is the best method of delivering the singleton frank or complete breech at term. The best mode of delivery for the pre-term breech is less clear. Vaginal breech delivery will be unavoidable in certain circumstances and it is therefore important to be adept with the techniques of vaginal breech delivery. The atraumatic technique of delivery of the baby presenting by the breech at times of Caesarean section is similar to that of assisted vaginal breech delivery. The number of vaginal breech deliveries is falling, and regular teaching using video clips or practising with mannequins will be necessary to preserve the skills of vaginal breech delivery. PMID- 11866496 TI - Twin delivery. AB - The incidence of twin pregnancy has increased worldwide over the past 10 years largely as a consequence of the assisted reproductive technologies. Issues such as intrapartum monitoring and operative interventions, especially with regard to the second twin, provide a unique challenge in labour and delivery. Epidemiological data suggest that the term twin has a threefold higher mortality rate than the singleton. It is the authors' view that many aspects of twin delivery deserve as much import as those features of twin gestations such as pre term birth and intrauterine growth restriction that, to date, have received much of the research and clinical interest in this area. Indications for elective Caesarean section are presented, incorporating new data derived from the delivery of the term singleton breech, and implications on the timing thereof are discussed. Vaginal delivery of both twins presenting by the vertex is recommended as safe as long as guidelines for the conduct of such delivery are followed. The recommended time interval between twins as well as the use of epidural, fetal monitoring and ultrasound in the delivery room are discussed. The second twin presenting as a non-vertex presents an urgent dilemma for accoucheurs. Data suggest that internal version and breech extractions are safer than external cephalic version provided that the appropriate techniques are applied. It is revealed, however, that the use of elective Caesarean section in this group of babies has not been subject to randomized controlled studies of sufficient power to determine the best method of delivery of the second twin - particularly in the low-birth-weight baby. PMID- 11866498 TI - Uterine rupture. AB - Uterine rupture is an uncommon obstetric event. It is important because it continues to be associated with maternal mortality, especially in developing countries, and with major maternal morbidity, particularly peripartum hysterectomy. It is also associated with a high incidence of perinatal mortality and morbidity worldwide. This chapter examines the incidence, aetiology, clinical presentation, complications and prevention of uterine rupture. The key factor in the cause of rupture is whether or not the uterus is scarred. Rupture of an unscarred uterus is rare, usually traumatic, and its incidence decreases with improvement in obstetric practice. Rupture of the scarred uterus is more common, and usually occurs after a trial of labour in a patient with a previous Caesarean section. This chapter also explores how the incidence and complications of uterine rupture may be minimized, and yet the incidence of vaginal birth after Caesarean section (VBAC) optimized, in clinical practice. PMID- 11866497 TI - Shoulder dystocia. AB - Shoulder dystocia is an uncommon but not rare obstetric emergency. Death of the infant is unusual but perinatal morbidity is frequent and can result in permanent injury. These cases carry significant medico-legal implications. This chapter covers the mechanisms, predisposing factors and management of shoulder dystocia. A well-rehearsed sequence of manoeuvres to manage shoulder dystocia will minimize fetal trauma. PMID- 11866499 TI - The surgical management of post-partum haemorrhage. AB - Life-threatening post-partum haemorrhage (PPH) occurs with a frequency of 1 per 1000 deliveries in the developed world. In the 1994-1996 Triennial Confidential Enquiry into Maternal Deaths in the United Kingdom primary PPH was responsible for five deaths. In this chapter we discuss briefly the assessment and initial medical management of the patient with primary PPH but concentrate on the surgical management where medical treatment has failed. The surgical management discussed includes both traditional or long-established management strategies together with newer, less radical surgical options, such as embolization techniques, uterine compression sutures and methods involving uterine tamponade, which are less hazardous to perform and have the advantage of preserving reproductive function. The recommendations of the reports from the Confidential Enquiries into Maternal Deaths in the UK are summarized at the end of the chapter. PMID- 11866500 TI - Lower genital tract and anal sphincter trauma. AB - Anal incontinence occurs more frequently in women but its incidence is grossly underestimated because of under-reporting. Obstetric trauma is a major cause of anal incontinence but it is only recently that attention has been focused on this subject. Episiotomy and choice of instrument at assisted delivery have been subjected to randomized trials but some issues, such as the benefit of episiotomy in instrumental delivery, have not been addressed. The management of acute anal sphincter rupture is inconsistent and, although studies report on the sub-optimal outcome, evidence-based guidelines are currently awaiting publication. Training in perineal anatomy and repair is poorly taught, and there is wide variation in classification of perineal tears. Consequently anal sphincter tears are being missed at delivery and/or inappropriately managed. This chapter aims to highlight these issues based on previous and current teaching and to recommend a protocol based on the best available evidence. PMID- 11866501 TI - Symphysiotomy and fetal destructive operations. AB - Symphysiotomy and fetal destructive operations - while rarely, if ever, performed in developed countries - are an option in developing nations of the world. Problems endemic to developing countries not infrequently predispose to patients arriving at health care facilities in a moribund state with neglected labour. The health care provider then has to decide on the options available to him to deliver the mother by the safest route without incurring morbidity and mortality. Under the circumstances, the outcome for the baby will depend on factors prevalent at the time. If the fetus is alive, then the choice is between a Caesarean section and symphysiotomy, and if the fetus has died, a destructive procedure is an option to abdominal-route delivery which carries considerable risk to the mother. The following review outlines the role of symphysiotomy and fetal destructive operations and their role in modern obstetrics. PMID- 11866506 TI - A small reservoir of disabled ORFs in the yeast genome and its implications for the dynamics of proteome evolution. AB - We surveyed the sequenced Saccharomyces cerevisiae genome (strain S288C) comprehensively for open reading frames (ORFs) that could encode full-length proteins but contain obvious mid-sequence disablements (frameshifts or premature stop codons). These pseudogenic features are termed disabled ORFs (dORFs). Using homology to annotated yeast ORFs and non-yeast proteins plus a simple region extension procedure, we have found 183 dORFs. Combined with the 38 existing annotations for potential dORFs, we have a total pool of up to 221 dORFs, corresponding to less than approximately 3% of the proteome. Additionally, we found 20 pairs of annotated ORFs for yeast that could be merged into a single ORF (termed a mORF) by read-through of the intervening stop codon, and may comprise a complete ORF in other yeast strains. Focussing on a core pool of 98 dORFs with a verifying protein homology, we find that most dORFs are substantially decayed, with approximately 90% having two or more disablements, and approximately 60% having four or more. dORFs are much more yeast-proteome specific than live yeast genes (having about half the chance that they are related to a non-yeast protein). They show a dramatically increased density at the telomeres of chromosomes, relative to genes. A microarray study shows that some dORFs are expressed even though they carry multiple disablements, and thus may be more resistant to nonsense-mediated decay. Many of the dORFs may be involved in responding to environmental stresses, as the largest functional groups include growth inhibition, flocculation, and the SRP/TIP1 family. Our results have important implications for proteome evolution. The characteristics of the dORF population suggest the sorts of genes that are likely to fall in and out of usage (and vary in copy number) in a strain-specific way and highlight the role of subtelomeric regions in engendering this diversity. Our results also have important implications for the effects of the [PSI+] prion. The dORFs disabled by only a single stop and the mORFs (together totalling 35) provide an estimate for the extent of the sequence population that can be resurrected readily through the demonstrated ability of the [PSI+] prion to cause nonsense-codon read-through. Also, the dORFs and mORFs that we find have properties (e.g. growth inhibition, flocculation, vanadate resistance, stress response) that are potentially related to the ability of [PSI+] to engender substantial phenotypic variation in yeast strains under different environmental conditions. (See genecensus.org/pseudogene for further information.) PMID- 11866507 TI - Amino acid activation of a dual-specificity tRNA synthetase is independent of tRNA. AB - Transfer RNA can play a role in amino acid activation by aminoacyl-tRNA synthetases. For the prolyl-tRNA synthetase (ProRS) of Methanococcus jannaschii, which activates both proline and cysteine, the role of tRNA in amino acid selection and activation is of interest in the effort to understand the mechanism of the dual-specificity. While activation of proline does not require tRNA, whether or not tRNA is required in the activation of cysteine has been a matter of debate. Here, investigation of a series of buffer conditions shows that activation of cysteine occurs without tRNA in a wide-range of buffers. However, the extent of cysteine activation is strongly buffer-dependent, varying over a 180-fold range. In contrast, the extent of proline activation is much less sensitive to buffer conditions, varying over only a 36-fold range. We also find that addition of tRNA has a small threefold stimulatory effect on cysteine activation. The lack of a major role of tRNA in activation of cysteine suggests that the dual-specificity enzyme must distinguish cysteine from proline directly, without the assistance of each cognate tRNA, to achieve the necessary specificity required for protein synthesis. PMID- 11866508 TI - Troponin-I interacts with the Met47 region of skeletal muscle actin. Implications for the mechanism of thin filament regulation by calcium. AB - Striated muscles are regulated by Ca(2+) via the thin filament proteins troponin (Tn) and tropomyosin (Tm). In the absence of Ca(2+), contraction is inhibited, whereas myosin-actin interaction and contraction can take place in its presence. Although it is well established that the interaction of troponin-I (TnI), the inhibitory subunit of Tn, with actin is required for the inhibition process and that there are two separate actin-binding regions in TnI that interact with actin, the molecular mechanism of this inhibition process is still not clear. Using TnI mutants with photocrosslinking probes attached to genetically engineered cysteine residues in each of the two actin-binding regions, we show that both regions are close to Met47 of actin in its outer domain. It has been proposed that the Ca(2+)-induced activation of contraction involves the movement of Tm from the outer to the inner domain of the actin filament. On the basis of our results presented here, we propose that the position of Tm at the outer domain of actin in the Ca(2+)-free state is stabilized by the presence of TnI over actin's outer domain via mutual interactions of all three components. In the presence of Ca(2+), TnI's actin-binding regions dissociate from actin allowing Tm to move toward actin's inner domain. PMID- 11866509 TI - Chiral mutagenesis of insulin's hidden receptor-binding surface: structure of an allo-isoleucine(A2) analogue. AB - The hydrophobic core of vertebrate insulins contains an invariant isoleucine residue at position A2. Lack of variation may reflect this side-chain's dual contribution to structure and function: Ile(A2) is proposed both to stabilize the A1-A8 alpha-helix and to contribute to a "hidden" functional surface exposed on receptor binding. Substitution of Ile(A2) by alanine results in segmental unfolding of the A1-A8 alpha-helix, lower thermodynamic stability and impaired receptor binding. Such a spectrum of perturbations, although of biophysical interest, confounds interpretation of structure-activity relationships. To investigate the specific contribution of Ile(A2) to insulin's functional surface, we have employed non-standard mutagenesis: inversion of side-chain chirality in engineered monomer allo-Ile(A2)-DKP-insulin. Although the analogue retains native structure and stability, its affinity for the insulin receptor is impaired by 50 fold. Thus, whereas insulin's core readily accommodates allo-isoleucine at A2, its activity is exquisitely sensitive to chiral inversion. We propose that the Ile(A2) side-chain inserts within a chiral pocket of the receptor as part of insulin's hidden functional surface. PMID- 11866511 TI - The reverse transcriptase of the R2 non-LTR retrotransposon: continuous synthesis of cDNA on non-continuous RNA templates. AB - R2 is a non-long terminal repeat (non-LTR) retrotransposon that inserts into the 28 S rRNA genes of arthropods. The element encodes two enzymatic activities: an endonuclease that specifically cleaves the 28 S gene target site, and a reverse transcriptase (RT) that can use the 3' end of the cleaved DNA to prime reverse transcription. R2 RT only utilizes RNA templates that contain the 3' untranslated region of the R2 element as templates in this target primed reverse transcription (TPRT) reaction. Here, detailed biochemical characterization of the R2 RT indicates that the enzyme is capable of making multiple, consecutive jumps between RNA templates. The terminal 3' nucleotide of the "acceptor" RNA and the 5' nucleotide of the "donor" RNA are frequently reverse transcribed in these jumps, indicating that the acceptor RNA does not anneal to the cDNA derived from the donor RNA template. These template jumps occur during TPRT as well as in non specific extension reactions in which reverse transcription is primed by an oligonucleotide annealed to the RNA template. Analysis of these RT assays done in the absence of the target DNA also revealed that the R2 RT can initiate reverse transcription near the 3' end of any RNA molecule using the 3' end of a second RNA molecule as primer. Again there is no requirement for sequence complementarity between the RNA used as template and the RNA used as primer. These properties of the R2 RT differ substantially from those of retroviral RTs but have similarities to the RT of the Mauriceville retroplasmid of Neurospora crassa. We present a model which relates these unusual properties of the R2 RT to structural differences from retroviral RTs as well as correlates these properties to the likely retrotransposition mechanism of R2 and other non-LTR retrotransposons. PMID- 11866510 TI - Defining the Bacillus subtilis sigma(W) regulon: a comparative analysis of promoter consensus search, run-off transcription/macroarray analysis (ROMA), and transcriptional profiling approaches. AB - The Bacillus subtilis extracytoplasmic function (ECF) sigma factor sigma(W) controls a large regulon that is strongly induced by alkali shock. To define the physiological role of sigma(W) we have sought to identify the complete set of genes under sigma(W) control. Previously, we described a promoter consensus search procedure to identify sigma(W) controlled genes. Herein, we introduce a novel method to identify additional target promoters: run-off transcription followed by macroarray analysis (ROMA). We compare the resulting list of targets with those identified in conventional transcriptional profiling studies and using the consensus search approach. While transcriptional profiling identifies genes that are strongly dependent on sigma(W) for in vivo expression, some sigma(W) dependent promoters are not detected due to the masking effects of other promoter elements, overlapping recognition with other ECF sigma factors, or both. Taken together, the consensus search, ROMA, and transcriptional profiling approaches establish a minimum of 30 promoter sites (controlling approximately 60 genes) as direct targets for activation by sigma(W). Significantly, no single approach identifies more than approximately 80% of the regulon so defined. We therefore suggest that a combination of two or more complementary approaches be employed in studies seeking to achieve maximal coverage when defining bacterial regulons. Our results indicate that sigma(W) controls genes that protect the cell against agents that impair cell wall biosynthesis but fail to reveal any connection to operons likely to function in adaptation to alkaline growth conditions. This is consistent with the observation that a sigW mutant is unaffected in its ability to survive alkali shock. We conclude that in B. subtilis sudden imposition of alkali stress activates the sigma(W) stress response, perhaps by impairing the ability of the cell wall biosynthetic machinery to function. PMID- 11866512 TI - The role of the zinc finger motif and of the residues at the amino terminus in the function of yeast ribosomal protein YL37a. AB - YL37a is an essential yeast ribosomal protein that has a C(2)-C(2) zinc finger motif. Replacement of the cysteine residues had yielded variants that lacked the capacity to bind zinc but still supported cell growth. In a continuation of an examination of the relation of the structure of YL37a to its function, the contribution of amino acid residues in the intervening sequence between the internal cysteine residues of the motif was evaluated. Substitutions of alanine for the lysine residues at positions 44, 45, or 48, or for arginine 49 slowed cell growth. The most severe effect was caused by a double-mutation, K48A-R49A. A mutation of tryptophan 55 to alanine was lethal. Mutations to alanine of six conserved residues (K6, K7, K13, Y14, R17, and Y18) in the amino-terminal region decreased cell growth; the Y14 mutation was lethal. An in vitro assay for binding of YL37a to individual 26 S rRNA domains was developed. Binding of the recombinant fusion protein MBP-YL37a was to domains II and III; the K(d) for binding to domain II was 79 nM; for domain III it was 198 nM. There was a close correspondence between the effect of mutations in YL37a on cell growth and on binding to 26 S rRNA. In the atomic structure of the 50 S subunit of Haloarcula marismortui, the archaebacteria homolog of yeast YL37a, L37ae, coordinates a zinc atom and the finger motif is folded and interacts mainly with domain III of 23 S rRNA; whereas the amino-terminal region of L37ae interacts primarily with domain II. The biochemical and genetic experiments complement the three-dimensional structure and define for the first time the functional importance of a subset of the residues in close proximity to nucleotides. PMID- 11866513 TI - The Saccharomyces cerevisiae histone acetyltransferase Gcn5 has a role in the photoreactivation and nucleotide excision repair of UV-induced cyclobutane pyrimidine dimers in the MFA2 gene. AB - How DNA repair enzymes or complexes gain access to chromatin is still not understood. Here, we have studied the role of the S. cerevisiae histone acetyltransferase Gcn5 in photoreactivation (PR) and nucleotide excision repair (NER) at the level of the genome, the MFA2 and RPB2 genes, and at specific nucleotides within MFA2. The deletion of GCN5 markedly reduced the PR and NER of UV-induced cyclobutane pyrimidine dimers in MFA2 but much less so in RPB2, whereas no detectable defect was seen for repair of the genome overall. In Delta(gcn5), the MFA2 mRNA level is reduced by fourfold, while transcription from RPB2 is reduced only to 80 %. These changes in transcription correlate with the changes in NER and PR found in the Delta(gcn5) mutant. However, changes in MFA2 transcription cannot account for the decrease in NER in the non-transcribed strand and the control region of MFA2 where global genome repair (GGR) operates. We conclude that the histone acetyltransferase Gcn5 influences PR and NER at MFA2 in both its transcribed and non-transcribed DNA, yet it has little effect on these processes for most of the yeast genome. As a result, we speculate that histone acetylation allows efficient access of the repair machinery to chromosomal DNA damages either indirectly via influencing transcription or directly via modifying chromatin structure irrespective of transcription. PMID- 11866514 TI - Architecture of Fis-activated transcription complexes at the Escherichia coli rrnB P1 and rrnE P1 promoters. AB - The transcription factor Fis activates the Escherichia coli rRNA promoters rrnB P1 and rrnE P1 by binding to sites centered at -71 and -72, respectively, and interacting with the C-terminal domain of the alpha subunit of RNA polymerase (RNAP alphaCTD). To understand the mechanism of activation by Fis at these promoters, we used oriented alpha-heterodimeric RNAPs and heterodimers of Fis to determine whether one or both subunits of alpha and Fis participate in the alphaCTD-Fis interaction. Our results imply that only one alphaCTD in the alpha dimer and only one activation-proficient subunit in the Fis dimer are required for activation by Fis. A library of alanine substitutions in alpha was used to identify the alphaCTD determinants required for Fis-dependent transcription at rrnB P1 and rrnE P1. We propose that the transcriptional activation region of the promoter-proximal subunit of the Fis dimer interacts with a determinant that includes E273 of one alphaCTD to activate transcription. We further suggest that the Fis contact to alphaCTD results in alphaCTD interactions with DNA that differ somewhat from those that occur at UP elements in the absence of Fis. The accompanying paper shows that the 273 determinant on alphaCTD is also targeted by Fis at the proP P2 promoter where the activator binds overlapping the -35 hexamer. Thus, similar Fis-alphaCTD interactions are used for activation of transcription when the activator is bound at very different positions on the DNA. PMID- 11866515 TI - The C-terminal domains of the RNA polymerase alpha subunits: contact site with Fis and localization during co-activation with CRP at the Escherichia coli proP P2 promoter. AB - Fis is a versatile transactivator that functions at many different promoters. Fis activates transcription at the RpoS-dependent proP P2 promoter when bound to a site that overlaps the minus sign35 hexamer by a mechanism that requires the C terminal domain of the alpha subunit of RNA polymerase (alphaCTD). The region on Fis responsible for activating transcription through the alphaCTD has been localized to a short beta-turn near the DNA-binding determinant on one subunit of the Fis homodimer. We report here that Fis-dependent activation of proP P2 transcription requires two discrete regions on the alphaCTD. One region, consisting of residues 264-265 and 296-297, mediates DNA binding. A second patch, comprising amino acid residues 271-273, forms a ridge on the surface of the alphaCTD that we propose interacts with Fis. The accompanying paper shows that these same regions on alphaCTD are utilized for transcriptional activation at the rrnB and rrnE P1 promoters by Fis bound to a site upstream of the core promoter (centered at minus sign71/minus sign72). In addition to stimulation of proP P2 transcription by Fis, CRP co-activates this promoter when bound to a remote site upstream from the promoter (centered at -121.5). RNA polymerase preparations lacking one alphaCTD of the alpha dimer were employed to demonstrate that the beta'-associated alpha(II)CTD was utilized preferentially by Fis at proP P2 in the presence and absence of CRP. These experiments define the overall architecture of the proP P2 initiation complex where Fis and CRP each function through a different alphaCTD. PMID- 11866516 TI - Thermodynamics of E. coli cytidine repressor interactions with DNA: distinct modes of binding to different operators suggests a role in differential gene regulation. AB - Interactions between the Escherichia coli cytidine repressor protein (CytR) and its operator sites at the different promoters that comprise the CytR regulon, play an important role in the regulation of these promoters. The natural operators are palindromes separated by variable length central spacers (0-9 bp). We have suggested that this variability affects the flexibility of CytR-DNA contacts, thereby affecting the critical protein-protein interactions between CytR and the cAMP receptor protein (CRP) that underlie differential repression and activation of CytR-regulated genes. To assess this hypothesis, we investigated the thermodynamics of CytR binding to the natural operator sequences found in udpP and deoP2. To separate effects due to spacing from effects due to the differing sequences of the recognition half-sites of these two operators, we also investigated CytR binding to artificial hybrid operators, in which the half site sequences of udpP and deoP2 were exchanged. Thermodynamic parameters, DeltaS(o), DeltaH(o) and DeltaC(o)(p), were determined by van't Hoff analysis of CytR binding, monitored by changes in the steady-state fluorescence anisotropy of dye-conjugated, operator-containing oligonucleotides. Large differences in thermodynamics were observed that depend primarily on the central spacer rather than the sequences of the recognition half-sites. Binding to operators with deoP2 spacing results in a very large, negative DeltaC(o)(p). Association is strongly favored enthalpically and strongly disfavored entropically at ambient temperature. By contrast, binding to operators with udpP spacing results in a small, negative DeltaC(o)(p). Association is weakly favored both enthalpically and entropically at ambient temperature. A difference of such magnitude in DeltaDeltaC(o)(p) has not been reported previously for specific binding of a transcription factor to different sites. The identical salt dependence of CytR binding to deoP2 and udpP operators indicates that ion-dependent processes do not contribute significantly to this difference. Thus, the different thermodynamic effects appear to reflect distinctly different modes of site-specific DNA binding. We discuss similarities to operator binding by CytR homologs among LacI family repressors, and we consider how different CytR binding modes might affect interactions with other components of the gene regulatory machinery that contribute to differential gene regulation. PMID- 11866517 TI - The large subunit of bacteriophage lambda's terminase plays a role in DNA translocation and packaging termination. AB - The DNA packaging enzyme of bacteriophage lambda, terminase, is a heteromultimer composed of a small subunit, gpNu1, and a large subunit, gpA, products of the Nu1 and A genes, respectively. The role of terminase in the initial stages of packaging involving the site-specific binding and cutting of the DNA has been well characterized. While it is believed that terminase plays an active role in later post-cleavage stages of packaging, such as the translocation of DNA into the head shell, this has not been demonstrated. Accordingly, we undertook a generalized mutagenesis of lambda's A gene and found ten lethal mutations, nine of which cause post-cleavage packaging defects. All were located in the amino terminal two-thirds of gpA, separate from the carboxy-terminal region where mutations affecting the protein's endonuclease activity have been found. The mutants fall into five groups according to their packaging phenotypes: (1) two mutants package part of the lambda chromosome, (2) one mutant packages the entire chromosome, but very slowly compared to wild-type, (3) two mutants do not package any DNA, (4) four mutants, though inviable, package the entire lambda chromosome, and (5) one mutant may be defective in both early and late stages of DNA packaging. These results indicate that gpA is actively involved in late stages of packaging, including DNA translocation, and that this enzyme contains separate functional domains for its early and late packaging activities. PMID- 11866518 TI - Extrusion of an imperfect palindrome to a cruciform in superhelical DNA: complete determination of energetics using a statistical mechanical model. AB - We present a detailed study of the extrusion of an imperfect palindrome, derived from the terminal regions of vaccinia virus DNA and contained in a superhelical plasmid, into a cruciform containing bulged bases. We monitor the course of extrusion by two-dimensional gel electrophoresis experiments as a function of temperature and linking number. We find that extrusion pauses at partially extruded states as negative superhelicity increases. To understand the course of extrusion with changes in linking number, DeltaLk, we present a rigorous semiempirical statistical mechanical analysis that includes complete coupling between DeltaLk, cruciform extrusion, formation of extrahelical bases, and temperature-dependent denaturation. The imperfections in the palindrome are sequentially incorporated into the cruciform arms as hairpin loops, single unpaired bases, and complex local regions containing several unpaired bases. We analyze the results to determine the free energies, enthalpies and entropies of formation of all local structures involved in extrusion. We find that, for each unpaired structure, the DeltaG, DeltaH and DeltaS of formation are all approximately proportional to the number of unpaired bases contained therein. This surprising result holds regardless of the arrangement or composition of unpaired bases within a particular structure. Imperfections have major effects on the overall energetics of cruciform extrusion and on the course of this transition. In particular, the extent of the DeltaLk change necessary to extrude an imperfect palindrome is considerably greater than that required for extrusion of the underlying perfect palindrome. Our analysis also suggests that, at higher temperatures, significant denaturation at the base of an imperfect cruciform can successfully compete with extension of the cruciform arms. PMID- 11866519 TI - Complete cDNA sequence of SAP-like pentraxin from Limulus polyphemus: implications for pentraxin evolution. AB - The serum amyloid P component (SAP)-like pentraxin Limulus polyphemus SAP is a recently discovered, distinct pentraxin species, of known structure, which does not bind phosphocholine and whose N-terminal sequence has been shown to differ markedly from the highly conserved N terminus of all other known horseshoe crab pentraxins. The complete cDNA sequence of Limulus SAP, and the derived amino acid sequence, the first invertebrate SAP-like pentraxin sequence, have been determined. Two sequences were identified that differed only in the length of the 3' untranslated region. Limulus SAP is synthesised as a precursor protein of 234 amino acid residues, the first 17 residues encoding a signal peptide that is absent from the mature protein. Phylogenetic analysis clusters Limulus SAP pentraxin with the horseshoe crab C-reactive proteins (CRPs) rather than the mammalian SAPs, which are clustered with mammalian CRPs. The deduced amino acid sequence shares 22% identity with both human SAP and CRP, which are 51% identical, and 31-35% with horseshoe crab CRPs. These analyses indicate that gene duplication of CRP (or SAP), followed by sequence divergence and the evolution of CRP and/or SAP function, occurred independently along the chordate and arthropod evolutionary lines rather than in a common ancestor. They further indicate that the CRP/SAP gene duplication event in Limulus occurred before both the emergence of the Limulus CRP variants and the mammalian CRP/SAP gene duplication. Limulus SAP, which does not exhibit the CRP characteristic of calcium-dependent binding to phosphocholine, is established as a pentraxin species distinct from all other known horseshoe crab pentraxins that exist in many variant forms sharing a high level of sequence homology. PMID- 11866520 TI - Functional analysis of a carboxyl-terminal phosphorylation mutant of the bovine papillomavirus E1 protein. AB - The papillomavirus E1 protein is essential for viral DNA replication, and phosphorylation of E1 appears to regulate protein function and DNA replication. Serine 584 of bovine papillomavirus E1 is in a conserved motif resembling a CK2 consensus site, and is phosphorylated by CK2 in vitro. Mutation of serine 584 to alanine eliminates replication of the viral genome in transient replication assays. Wild-type and mutant E1 proteins were expressed from recombinant baculoviruses and used to assess biochemical functions of the amino acid 584 substitution. Helicase enzyme activity, E1 binding to the viral E2 protein and to cellular DNA polymerase alpha-primase were all unaffected in the mutant protein. Binding of E1 to viral replication origin DNA sequences was reduced in the mutant, but not eliminated. The carboxyl-terminal region of the protein appears to play a role in regulating E1 function, and adds to a complex picture emerging for papillomavirus DNA replication control. PMID- 11866521 TI - Effects of temperature on the dynamic behaviour of the HIV-1 nucleocapsid NCp7 and its DNA complex. AB - The nucleocapsid protein NCp7 of human immunodeficiency virus type 1 (HIV-1) contains two highly conserved CCHC zinc fingers and is involved in many crucial steps of the virus life-cycle. A large number of physiological roles of NCp7 involve its binding to single-stranded nucleic acid chains. Several solution structures of NCp7 and its complex with single-stranded RNA or DNA have been reported. We have investigated the changes in the dynamic behaviour experienced by the (12-53)NCp7 peptide upon DNA binding using (15)N heteronuclear relaxation measurements at 293 K and 308 K, and fluorescence spectroscopy. The relaxation data were interpreted using the reduced spectral density approach, which allowed the high-frequency motion, overall tumbling rates and the conformational exchange contributions to be characterized for various states of the peptide without using a specific motional model. Analysis of the temperature-dependent correlation times derived from both NMR and fluorescence data indicated a co-operative change of the molecular shape of apo (12-53)NCp7 around 303 K, leading to an increased hydrodynamic radius at higher temperatures. The binding of (12-53)NCp7 to a single-stranded d(ACGCC) pentanucleotide DNA led to a reduction of the conformational flexibility that characterized the apo peptide. Translational diffusion experiments as well as rotational correlation times indicated that the (12-53)NCp7/d(ACGCC) complex tumbles as a rigid object. The amplitudes of high frequency motions were restrained in the complex and the occurrence of conformational exchange was displaced from the second zinc finger to the linker residue Ala30. PMID- 11866522 TI - Concerted evolution between duplicated genetic elements in Helicobacter pylori. AB - The Helicobacter pylori genome includes a family of outer membrane proteins (OMPs) with substantial N and C-terminal identity. To better understand their evolution, the nucleotide sequences for two members, babA and babB, were determined from a worldwide group of 23 strains. The geographic origin of each strain was found to be the major determinant of phylogenetic structure, with strains of Eastern and Western origin showing greatest divergence. For strains 96 10 (Japan) and 96-74 (USA), the 5' regions of babB are replaced with babA sequences, demonstrating that recombination occurs between the two loci. babA and babB have nearly equivalent variation in nucleotide and amino acid identity, and frequencies of synonymous and non-synonymous substitutions. Both genes have segmental conservation but within the 3' segment, substitution patterns are nearly identical. Although babA and babB 5' and midregion segment phylogenies show strong interstrain similarity, the 3' segments show strong intrastrain similarity, indicative of concerted evolution. Within these 3' segments, the lower intrastrain than interstrain frequencies of nucleotide substitutions, which are below mean background H. pylori substitution frequencies, indicate selection against intrastrain diversification. Since babA/babB gene conversions likely underlie the concerted evolution of the 3' segments, in an experimental system, we demonstrate that gene conversions can frequently (10(-3)) occur in H. pylori. That these events are recA-dependent and DNase-resistant indicates their likely cause is intragenomic recombination. PMID- 11866523 TI - Analysis of shuffled gene libraries. AB - In vitro recombination of homologous genes (family shuffling) has been proposed as an effective search strategy for laboratory evolution of genes and proteins. Few data are available, however, on the composition of shuffled gene libraries, from which one could assess the efficiency of recombination and optimize protocols. Here, probe hybridization is used in a macroarray format to analyze chimeric DNA libraries created by DNA shuffling. Characterization of hundreds of shuffled genes encoding dioxygenases has elucidated important biases in the shuffling reaction. As expected, crossovers are favored in regions of high sequence identity. A sequence-based model of homologous recombination that captures this observed bias was formulated using the experimental results. The chimeric genes were found to show biases in the incorporation of sequences from certain parents, even before selection. Statistically different patterns of parental incorporation in genes expressing functional proteins can help to identify key sequence-function relationships. PMID- 11866524 TI - Protein translocase of the outer mitochondrial membrane: role of import receptors in the structural organization of the TOM complex. AB - The mitochondrial outer membrane contains a multi-subunit machinery responsible for the specific recognition and translocation of precursor proteins. This translocase of the outer membrane (TOM) consists of three receptor proteins, Tom20, Tom22 and Tom70, the channel protein Tom40, and several small Tom proteins. Single-particle electron microscopy analysis of the Neurospora TOM complex has led to different views with two or three stain-filled centers resembling channels. Based on biochemical and electron microscopy studies of the TOM complex isolated from yeast mitochondria, we have discovered the molecular reason for the different number of channel-like structures. The TOM complex from wild-type yeast contains up to three stain-filled centers, while from a mutant yeast selectively lacking Tom20, the TOM complex particles contain only two channel-like structures. From mutant mitochondria lacking Tom22, native electrophoresis separates an approximately 80 kDa subcomplex that consists of Tom40 only and is functional for accumulation of a precursor protein. We conclude that while Tom40 forms the import channels, the two receptors Tom22 and Tom20 are required for the organization of Tom40 dimers into larger TOM structures. PMID- 11866525 TI - Analysis of Rous sarcoma virus capsid protein variants assembled on lipid monolayers. AB - During assembly and morphogenesis of Rous sarcoma virus (RSV), proteolytic processing of the structural precursor (Pr76Gag) protein generates three capsid (CA) protein variants, CA476, CA479, and CA488. The proteins share identical N terminal domains (NTDs), but are truncated at residues corresponding to gag codons 476, 479, and 488 in their CA C-terminal domains (CTDs). To characterize oligomeric forms of the RSV CA variants, we examined 2D crystals of the capsid proteins, assembled on lipid monolayers. Using electron microscopy and image analysis approaches, the CA proteins were observed to organize in hexagonal (p6) arrangements, where rings of membrane-proximal NTD hexamers were spaced at 95 A intervals. Differences between the oligomeric structures of the CA variants were most evident in membrane-distal regions, where apparent CTDs interconnect hexamer rings. In this region, CA488 connections were observed readily, while CA476 and CA479 contacts were resolved poorly, suggesting that in vivo processing of CA488 to the shorter forms may permit virions to adopt a dissembly-competent conformation. In addition to crystalline arrays, the CA479 and CA488 proteins formed small spherical particles with diameters of 165-175 A. The spheres appear to be arranged from hexamer or hexamer plus pentamer ring subunits that are related to the 2D crystal forms. Our results implicate RSV CA hexamer rings as basic elements in the assembly of RSV virus cores. PMID- 11866526 TI - Crystal structure of activated ribulose-1,5-bisphosphate carboxylase/oxygenase from green alga Chlamydomonas reinhardtii complexed with 2-carboxyarabinitol-1,5 bisphosphate. AB - Ribulose-1,5-bisphosphate carboxylase/oxygenase (rubisco) catalyzes the initial steps of photosynthetic carbon reduction and photorespiratory carbon oxidation cycles by combining CO(2) and O(2), respectively, with ribulose-1,5-bisphosphate. Many photosynthetic organisms have form I rubiscos comprised of eight large (L) and eight small (S) subunits. The crystal structure of the complex of activated rubisco from the green alga Chlamydomonas reinhardtii and the reaction intermediate analogue 2-carboxyarabinitol-1,5-bisphosphate (2-CABP) has been solved at 1.84 A resolution (R(cryst) of 15.2 % and R(free) of 18.1 %). The subunit arrangement of Chlamydomonas rubisco is the same as those of the previously solved form I rubiscos. Especially, the present structure is very similar to the activated spinach structure complexed with 2-CABP in the L-subunit folding and active-site conformation, but differs in S-subunit folding. The central insertion of the Chlamydomonas S-subunit forms the longer betaA-betaB loop that protrudes deeper into the solvent channel of rubisco than higher plant, cyanobacterial, and red algal (red-like) betaA-betaB loops. The C-terminal extension of the Chlamydomonas S-subunit does not protrude into the solvent channel, unlike that of the red algal S-subunit, but lies on the protein surface anchored by interactions with the N-terminal region of the S-subunit. Further, the present high-resolution structure has revealed novel post-translational modifications. Residue 1 of the S-subunit is N(alpha)-methylmethionine, residues 104 and 151 of the L-subunit are 4-hydroxyproline, and residues 256 and 369 of the L-subunit are S(gamma)-methylcysteine. Furthermore, the unusual electron density of residue 471 of the L-subunit, which has been deduced to be threonine from the genomic DNA sequence, suggests that the residue is isoleucine produced by RNA editing or O(gamma)-methylthreonine. PMID- 11866527 TI - X-ray diffraction of heavy-atom labelled two-dimensional crystals of rhodopsin identifies the position of cysteine 140 in helix 3 and cysteine 316 in helix 8. AB - We have used site-specific heavy-atom labelling and X-ray diffraction to localize single amino acid residues in the cytoplasmic domain of the integral membrane protein rhodopsin, the dim-light photoreceptor of retinal vertebrate rod cells. Two-dimensional orthorhombic crystals of the space group p22(1)2(1) (a=59.5(+/-1) A and b=82.7(+/-1.5) A) were produced from detergent-solubilized, partially delipidated rhodopsin. To obtain milligram amounts of two-dimensional crystals, which are required for X-ray diffraction, the yield of the crystalline material was significantly increased by reconstitution of rhodopsin in the presence of cholesterol (1:2 to 1:10 mol/mol) and by adding polar organic solvents to the dialysis buffer. The native cysteine residues C140 and C316 were then selectively labelled with mercury using the sulphydryl-specific reagent p chloromercuribenzoate (1.6-2.1 mol Hg per mol rhodopsin). The labelling did not affect the unit cell dimensions. Optical absorption spectra of labelled and native two-dimensional rhodopsin crystals showed the characteristic 11-cis retinal peak at 498 nm, which corresponds to the dark state of rhodopsin. The in plane position of the mercury label was calculated at 9.5 A resolution from the intensity differences in the X-ray diffraction patterns of labelled and native crystals using Fourier difference methods and the phase information from electron crystallography. The label positions were in excellent agreement with the positions of C140 at the cytoplasmic end of helix 3 and of C316 in the cytoplasmic helix 8 recently obtained from three-dimensional rhodopsin crystals. Whereas these high-resolution diffraction studies were performed under cryogenic conditions (100 K), our results were obtained at room temperature with fully hydrated membranes and in the absence of loop-loop crystal contacts. To study the structural changes of the cytoplasmic loops involved in activation and signal transduction, our more physiological conditions offer important advantages. Furthermore, the localization of C316 is the first direct proof that the electron density on top of helix 1 observed by cryo-electron microscopy is a part of the C terminal loop. Our approach is of particular interest for investigations of other membrane proteins, for which 3D crystals are not available. Structural constraints from heavy-atom labels at strategic sites enable the assignment of a position in the amino acid sequence to features visible in a low-resolution density map and the study of conformational changes associated with different functional states of the membrane protein. PMID- 11866528 TI - Crystallographic studies on human BST-1/CD157 with ADP-ribosyl cyclase and NAD glycohydrolase activities. AB - cADPR is the novel second messenger that elicits calcium release from intracellular calcium stores and works independently of IP(3). In mammals, the ADP-ribosyl cyclase function is found in two membrane proteins, CD38 and BST 1/CD157. These enzymes, exposed extracellularly, bear cADPR hydrolase and NAD glycohydrolase activities. In spite of its functional importance, the structural basis of these enzymatic reactions remains elusive. We determined the crystal structures of the extracellular region of human BST-1 at atomic resolution in the free form and in complexes with five substrate analogues: nicotinamide, NMN, ATPgammaS, ethenoNADP, and ethenoNAD. The three-dimensional structural views of the reaction centre with these ligands revealed the mode of substrate binding and the catalytic mechanism of the multifunctional enzymatic reactions. In each catalytic cleft of the dimeric enzyme, substrates are recognized predominantly through van der Waals interactions with two tryptophan residues, and thereby the N-glycosidic bond of NAD is correctly exposed near a catalytic glutamate residue. Its carboxyl side-chain stabilizes the catalytic intermediate of the S(N)-1 type reaction. This conformation of the catalytic cleft also implies the mechanism of cyclization between the adenine base and the ribose. The three key residues are invariant among the sequences of BST-1, CD38, and Aplysia cyclase, and hence this substrate recognition mode and catalytic scheme appear to be common in the cyclase family. PMID- 11866529 TI - Crystal structure of the 30 S ribosomal subunit from Thermus thermophilus: structure of the proteins and their interactions with 16 S RNA. AB - We present a detailed analysis of the protein structures in the 30 S ribosomal subunit from Thermus thermophilus, and their interactions with 16 S RNA based on a crystal structure at 3.05 A resolution. With 20 different polypeptide chains, the 30 S subunit adds significantly to our data base of RNA structure and protein RNA interactions. In addition to globular domains, many of the proteins have long, extended regions, either in the termini or in internal loops, which make extensive contact to the RNA component and are involved in stabilizing RNA tertiary structure. Many ribosomal proteins share similar alpha+beta sandwich folds, but we show that the topology of this domain varies considerably, as do the ways in which the proteins interact with RNA. Analysis of the protein-RNA interactions in the context of ribosomal assembly shows that the primary binders are globular proteins that bind at RNA multihelix junctions, whereas proteins with long extensions assemble later. We attempt to correlate the structure with a large body of biochemical and genetic data on the 30 S subunit. PMID- 11866531 TI - Three topologically equivalent core residues affect the transition state ensemble in a protein folding reaction. AB - The single domain protein, interleukin-1beta, is representative of a distinct class of proteins characterized by their beta-trefoil topology. Each subdomain of this structural class is composed of a beta beta beta loop beta (betabetabetaLbeta) motif comprised of approximately 50 residues and gives the protein a pseudo- 3-fold axis of symmetry. A common feature of proteins in this topological family appears to be that they are slow folders, which reach the native state on the order of tens to 100s of seconds. Sequence analysis of interleukin-1beta indicates that three phenylalanine residues located at positions 42, 101, and 146 are well conserved, separated by approximately 50 residues in the primary sequence, located in similar positions in the pseudo symmetric units of the trefoil, and are juxtaposed to one another in conformational space. These residues surround the hydrophobic cavity and "pin" the hairpin triplet cap to the core beta-barrel. To determine if cap-barrel interactions are involved in maintaining the structural stability and cooperativity or in controlling the slow formation of the native state, we performed a series of mutational studies. The results indicate that interleukin 1beta tolerates large increases in side-chain volume at these three topologically conserved sites with little effect on stability, while the kinetics show significant differences in both the unfolding and refolding rates. Taken together, our results indicate that these conserved core residues are essential contacts in the transition-state ensemble for folding. PMID- 11866530 TI - Solution structure of a phage-derived peptide antagonist in complex with vascular endothelial growth factor. AB - Vascular endothelial growth factor (VEGF) is a potent endothelial cell-specific mediator of angiogenesis and vasculogenesis. VEGF is involved pathologically in cancer, proliferative retinopathy and rheumatoid arthritis, and as such represents an important therapeutic target. Three classes of disulfide constrained peptides that antagonize binding of the VEGF dimer to its receptors, KDR and Flt-1, were identified previously using phage display methods. NMR studies of a representative peptide from the most potent class of these peptide antagonists, v107 (GGNECDAIRMWEWECFERL), were undertaken to characterize its interactions with VEGF. v107 has no defined structure free in solution, but binding to VEGF induces folding of the peptide. The solution structure of the VEGF receptor-binding domain-v107 complex was determined using 3940 (1970 per VEGF monomer) internuclear distance and 476 (238 per VEGF monomer) dihedral angle restraints derived from NMR data obtained using samples containing either (13)C/(15)N-labeled protein plus excess unlabeled peptide or (13)C/(15)N-labeled peptide plus excess unlabeled protein. Residual dipolar coupling restraints supplemented the structure determination of the complex and were found to increase significantly both the global precision of VEGF in the complex and the agreement with available crystal structures of VEGF. The calculated ensemble of structures is of high precision and is in excellent agreement with the experimental restraints. v107 has a turn-helix conformation with hydrophobic residues partitioned to one face of the peptide and polar or charged residues at the other face. Contacts between two v107 peptides and the VEGF dimer are mediated by primarily hydrophobic side-chain interactions. The v107-binding site on VEGF overlaps partially with the binding site of KDR and is similar to that for domain 2 of Flt-1. The structure of the VEGF-v107 complex provides new insight into how binding to VEGF can be achieved that may be useful for the design of small molecule antagonists. PMID- 11866532 TI - Motifs of serine and threonine can drive association of transmembrane helices. AB - Known sequence motifs containing key glycine residues can drive the homo oligomerization of transmembrane helices. To find other motifs, a randomized library of transmembrane interfaces was generated in which glycine was omitted. The TOXCAT system, which measures transmembrane helix association in the Escherichia coli inner membrane, was used to select high-affinity homo oligomerizing sequences in this library. The two most frequently occurring motifs were SxxSSxxT and SxxxSSxxT. Isosteric mutations of any one of the serine and threonine residues to non-polar residues abolished oligomerization, indicating that the interaction between these positions is specific and requires an extended motif of serine and threonine hydroxyl groups. Computational modeling of these sequences produced several chemically plausible structures that contain multiple hydrogen bonds between the serine and threonine residues. While single serine or threonine side-chains do not appear to promote helix association, motifs can drive strong and specific association through a cooperative network of interhelical hydrogen bonds. PMID- 11866533 TI - Differences between the prion protein and its homolog Doppel: a partially structured state with implications for scrapie formation. AB - The key event in the pathogenesis of prion diseases is a conformational change in the prion protein (PrP). Models for conversion of PrP(C) into PrP(Sc) typically implicate an, as yet, unidentified intermediate. In an attempt to identify such an intermediate, we used native-state hydrogen exchange monitored with NMR. Although we were unable to detect an intermediate directly, we observed substantial protection above that expected based upon measurements of the global stability of PrP (>2 kcal mol(-1) super protection). This super protection implicates either structure in the denatured state or the presence of an intermediate. Similar experiments with Doppel, a homolog of PrP that does not form infectious prions, failed to demonstrate such super protection. This suggests that the partially structured state of PrP encompassing portions of the B and C helices, may be a significant factor in the ability of PrP to convert from PrP(C) to PrP(Sc). PMID- 11866534 TI - XKCM1 acts on a single protofilament and requires the C terminus of tubulin. AB - The stability of microtubules during the cell-cycle is regulated by a number of cellular factors, some of which stabilize microtubules and others that promote breakdown. XKCM1 is a kinesin-like protein that induces microtubule depolymerization and is required for mitotic spindle assembly. We have examined the binding and depolymerization effects of XKCM1 on different tubulin polymers in order to learn about its mechanism of action. Zinc-induced tubulin polymers, characterized by an anti-parallel protofilament arrangement, are depolymerized by XKCM1, indicating that this enzyme acts on a single protofilament. GDP-tubulin rings, which correspond to the low-energy state of tubulin, are stable only under conditions that inhibit XKCM1 depolymerizing activity, but can be stabilized by XKCM1 bound to AMPPNP. Tubulin polymers made of subtilisin-treated tubulin (lacking the tubulin C-terminal tail) are resistant to XKCM1-induced depolymerization, suggesting that the interaction of the acidic tail of tubulin with basic residues in XKCM1 unique to Kin I proteins is required for depolymerization. PMID- 11866535 TI - Reaction mechanism of GTP cyclohydrolase I: single turnover experiments using a kinetically competent reaction intermediate. AB - GTP cyclohydrolase I catalyses the transformation of GTP into dihydroneopterin 3' triphosphate, which is the first committed precursor of tetrahydrofolate and tetrahydrobiopterin. The kinetically competent reaction intermediate, 2-amino-5 formylamino-6-ribosylamino-4(3H)-pyrimidinone, was used as substrate for single turnover experiments monitored by multiwavelength photometry. The early reaction phase is characterized by the rapid appearance of an optical transient with an absorption maximum centred at 320. This species is likely to represent a Schiff base intermediate at the initial stage of the Amadori rearrangement of the carbohydrate side-chain. Deconvolution of the optical spectra suggested four linearly independent processes. A fifth reaction step was attributed to photodecomposition of the enzyme product. Pre-steady state experiments were also performed with the H179A mutant which can catalyse a reversible conversion of GTP to 2-amino-5-formylamino-6-ribosylamino-4(3H)-pyrimidinone but is unable to form the final product, dihydroneopterin triphosphate. Optical spectroscopy failed to detect any intermediate in the reversible reaction sequence catalysed by the mutant protein. The data obtained with the wild-type and mutant protein in conjunction with earlier quenched flow studies show that the enzyme-catalysed opening of the imidazole ring of GTP and the hydrolytic release of formate from the resulting formamide type intermediate are both rapid reactions by comparison with the subsequent rearrangement of the carbohydrate side-chain which precedes the formation of the dihydropyrazine ring of dihydroneopterin triphosphate. PMID- 11866536 TI - SnapDRAGON: a method to delineate protein structural domains from sequence data. AB - We describe a method to identify protein domain boundaries from sequence information alone based on the assumption that hydrophobic residues cluster together in space. SnapDRAGON is a suite of programs developed to predict domain boundaries based on the consistency observed in a set of alternative ab initio three-dimensional (3D) models generated for a given protein multiple sequence alignment. This is achieved by running a distance geometry-based folding technique in conjunction with a 3D-domain assignment algorithm. The overall accuracy of our method in predicting the number of domains for a non-redundant data set of 414 multiple alignments, representing 185 single and 231 multiple domain proteins, is 72.4 %. Using domain linker regions observed in the tertiary structures associated with each query alignment as the standard of truth, inter domain boundary positions are delineated with an accuracy of 63.9 % for proteins comprising continuous domains only, and 35.4 % for proteins with discontinuous domains. Overall, domain boundaries are delineated with an accuracy of 51.8 %. The prediction accuracy values are independent of the pair-wise sequence similarities within each of the alignments. These results demonstrate the capability of our method to delineate domains in protein sequences associated with a wide variety of structural domain organisation. PMID- 11866537 TI - Determinants in nuclease specificity of Ape1 and Ape2, human homologues of Escherichia coli exonuclease III. AB - Abasic sites and non-conventional 3'-ends, e.g. 3'-oxidized fragments (including 3'-phosphate groups) and 3'-mismatched nucleotides, arise at significant frequency in the genome due to spontaneous decay, oxidation or replication errors. To avert the potentially mutagenic or cytotoxic effects of these chromosome modifications/intermediates, organisms are equipped with apurinic/apyrimidinic (AP) endonucleases and 3'-nucleases that initiate repair. Ape1, which shares homology with Escherichia coli exonuclease III (ExoIII), is the major abasic endonuclease in mammals and an important, yet selective, contributor to 3'-end processing. Mammals also possess a second protein (Ape2) with sequence homology to ExoIII, but this protein exhibits comparatively weak AP site-specific and 3'-nuclease activities. Prompted by homology modeling studies, we found that substitutions in the hydrophobic pocket of Ape1 (comprised of F266, W280 and L282) reduce abasic incision potency about fourfold to 450,000-fold, while introduction of an ExoIII-like pocket into Ape2 enhances its AP endonuclease function. We demonstrate that mutations at F266 and W280 of Ape1 increase 3' to 5' DNA exonuclease activity. These results, coupled with prior comparative sequence analysis, indicate that this active-site hydrophobic pocket influences the substrate specificity of a diverse set of sequence-related proteins possessing the conserved four-layered alpha/beta-fold. Lastly, we report that wild-type Ape1 excises 3'-mismatched nucleotides at a rate up to 374-fold higher than correctly base-paired nucleotides, depending greatly on the structure and sequence of the DNA substrate, suggesting a novel, selective role for the human protein in 3'-mismatch repair. PMID- 11866538 TI - Vascular endothelial growth factor modulates the Tie-2:Tie-1 receptor complex. AB - The receptor tyrosine kinase Tie-1 is expressed predominantly in endothelial cells where it physically associates with the related receptor Tie-2. Positive signalling through Tie-2 is associated with microvessel stability and suppression of this signal is thought to be required for vascular endothelial growth factor (VEGF)-induced microvessel remodelling or growth. Here we examine the effects of VEGF on Tie-1 and the Tie-2:Tie-1 complex. We show that VEGF induces generation of the Tie-1 endodomain and loss of the full-length receptor. The effects of VEGF on endodomain formation are not suppressed by inhibitors of protein kinase C and do not involve the nitric oxide signalling pathway. Tyrosine kinase inhibitors, in contrast, do abolish endodomain generation in response to the endothelial growth factor. VEGF stimulation of cells does not cause dissociation of the Tie 2:Tie-1 complex; rather the complex is converted to a form comprising the full length-Tie-2 and Tie-1 endodomain. VEGF can therefore switch the Tie-2:Tie-1 complex between two different forms in endothelial cells. The ability of VEGF to modulate Tie-1 and the Tie-2:Tie-1 complex provides a mechanism whereby this initiator of vessel growth and remodelling can directly modulate receptors involved in vessel stabilization. Such cross-talk is likely to be important in the coordinate control of blood vessel formation during development and in postnatal angiogenesis. PMID- 11866539 TI - Identification of endothelial cell genes expressed in an in vitro model of angiogenesis: induction of ESM-1, (beta)ig-h3, and NrCAM. AB - Blood vessel growth by angiogenesis plays an essential role in embryonic development, wound healing, and tumor growth. To understand the molecular cues underlying this process we have used the PCR-based subtractive hybridization method, representational difference analysis, to identify genes upregulated in endothelial cells (EC) forming tubes in 3D collagen gels, compared to migrating and proliferating cells in 2D cultures. We identified several previously characterized angiogenic markers, including the alpha(v) chain of the alpha(v)beta3 integrin and plasminogen activator inhibitor-1, suggesting overlap in gene expression between tube-forming cells in vitro and in vivo. We also found a 2- to 10-fold upregulation of (beta)ig-h3 (a collagen-binding extracellular matrix protein), NrCAM (a "neural" cell adhesion molecule), Annexin II (a tPA receptor), ESM-1 (an EC-specific molecule of unknown function), and Id2 (an inhibitory bHLH transcription factor). We identified a novel splice variant of the ESM-1 gene and also detected dramatically enhanced expression of ESM-1 and (beta)ig-h3 in several tumors. Antisense oligonucleotides to (beta)ig-h3 blocked both gene expression and tube formation in vitro, suggesting that (beta)ig-h3 may play a critical role in EC-matrix interactions. These data expand the suite of genes implicated in vascular remodeling and angiogenesis. PMID- 11866541 TI - Arterial blood flow and microcirculatory changes in a rat groin flap after thrombosis induced by electrical stimulation of the artery. AB - An experimental model of acute arterial thrombosis was developed in a rat groin flap model. Electrical stimulation was delivered to the flap artery while measuring blood flow in the artery and in the flap microcirculation using a laser Doppler system. Electrical stimulation produced an occlusive thrombosis in 10 rats within 68.1 +/- 19.26 (mean +/- SE) min. Thrombosis formation produced a rapid decrease of the LDF readings in the artery (90%) and in the flap (70%), the decrease being fairly parallel. Following the spontaneous thrombolysis, the artery perfusion recovered to baseline level but the flap blood flow only recovered by 10-20%. During electrical stimulation there was no change of the systemic blood pressure. The dynamic course of thrombus formation was documented on a videotape recorder through a microscope-mounted video camera and monitored on a television screen. Segments of the artery were obtained during and at the end of the experiments. The histological examination revealed arterial thrombosis composed of red blood cells, fibrin, and white blood cells, without any significant architectural and endothelial changes in the vessel walls. The study suggests that this model using electric vessel stimulation is effective for inducing arterial thrombosis and provides a simple method for recording the dynamic course of thrombus formation. PMID- 11866540 TI - p42/44MAPK regulates baseline permeability and cGMP-induced hyperpermeability in endothelial cells. AB - We tested the hypothesis that p42/44MAPK and p38MAPK (mitogen-activated protein kinases; MAPK) signaling pathways regulate endothelial cell permeability to macromolecules. Passage 2-4 human umbilical vein endothelial cells (HUVEC) were grown to confluence on fibronectin-coated Snapwell membranes. The flux of fluorescein isothiocyanate-labeled dextran-70 across the HUVEC monolayers served to determine permeability. Application of 1 mM 8-bromo 3' 5'-cyclic guanosine monophosphate (8-Br-cGMP) increased permeability from 7.0 +/- 1.6 x 10(-6) to 12.5 +/- 2.8 x 10(-6) cm/s (P < 0.05). Pretreatment of HUVEC for 60 min with a selective p42/44MAPK inhibitor (AG126 at 2.7 and 27 microM) blocked 8-Br-cGMP induced hyperpermeability. However, inhibition of p38MAPK (SB203580 at 0.6 microM) did not influence the cGMP-induced hyperpermeability response. AG126, administered at 27 microM, decreased baseline permeability from 7.9 +/- 0.5 x 10( 6) to 5.9 +/- 0.5 x 10(-6) cm/s (P < 0.05). Our results indicate that the p42/44MAPK signaling pathway is important in the regulation of baseline permeability and cGMP-induced hyperpermeability. PMID- 11866542 TI - Advanced glycation end products induce angiogenesis in vivo. AB - Advanced glycation end products (AGEs) have been thought to participate in diabetic microangiopathy. However, the effects of AGEs on angiogenesis have so far been mainly examined either in vitro or by using cultured cells. In the present study, we have analyzed whether AGEs induce angiogenesis in vivo by using the chorioallantoic membrane (CAM) assay. The CAM assay was carried out in embryonated hen eggs to determine the effects of AGEs. Following generation of AGEs based on bovine serum albumin (BSA), either AGE-BSA or nonglycated BSA was administered to the CAM and their effects on angiogenesis were assessed, together with an inhibitory effect of an anti-AGE antibody against AGE-BSA-induced angiogenesis. The histological features of AGE-induced vascular lumens were examined by immunohistochemical analysis for Factor VIII and smooth muscle alpha actin. AGE-BSA induced angiogenesis in CAM in a dose- and time-dependent manner. AGE-induced angiogenesis on CAM was neutralized by the anti-AGE antibody. Immunohistochemical analysis demonstrated that AGE-induced vascular lumens were devoid of pericytes. Our data demonstrated that AGEs are an angiogenetic factor and that our system of AGE-induced abnormal vessels in CAMs is useful in further investigations of the mechanism of diabetic retinal angiogenesis and can also be used to provide a therapeutic model for diabetic angiopathy. PMID- 11866543 TI - Distinction between atropine-sensitive control of microvascular and cardiac oscillatory activity. AB - This study tested whether "high frequency" oscillations (HF; >0.12 cycles/s) in microvascular flow during a vasoconstrictive challenge constitute a homeostatic cholinergic response at the level of the microvasculature or simply represent oscillations that originate at the heart and are transmitted to passive microvascular beds. Heart rate, blood pressure, respiratory rate, and laser Doppler flowmetry of the forehead and finger were monitored in healthy volunteers at baseline, during systemic infusion of phenylephrine (0.4-0.6 microg/kg/min), and during subsequent addition of intravenous atropine (< or = 2.0 mg/70 kg). Spectral-domain analysis documented that atropine-sensitive oscillatory power of the R-wave to R-wave intervals of the electrocardiogram was predominant at the respiratory frequency (0.20 Hz) at baseline and during phenylephrine infusion. In contrast, arteriolar-capillary networks of the forehead developed a prominent atropine-sensitive oscillatory peak at 0.14 +/- 0.02 Hz during phenylephrine infusion (P < 0.05 for differences in oscillatory magnitude and frequency between forehead flow and R-R intervals). The cross-power spectral density confirmed the lack of common power between forehead flow and R-R oscillations. Post hoc assessments showed that---similar to heart rate---systemic pressure and systemic flow also had persistent power at 0.20 Hz and did not develop a peak at the forehead oscillatory frequency; phenylephrine likewise did not induce atropine sensitive oscillations in the finger, a finding attributable to adrenergic predominance in this region. We conclude that atropine-sensitive oscillatory activity in the forehead microvasculature in response to a vasoconstrictive challenge constitutes a local response that is not due to, nor associated with, mechanical transmission from the heart and proximal vasculature. PMID- 11866544 TI - Shear-induced changes in endothelin-1 secretion of microvascular endothelial cells. AB - Human glomerular microvascular endothelial cell (HGMEC) culture monolayers were maintained in static culture as controls or subjected to steady laminar shear stress of 0.5, 1.0, or 1.5 N/m2. Over 25 h of shear, the cumulative secretion of ET-1 was 705.4 pg/cm2 in the control, 820.7 pg/cm2 at 0.5 N/m2, 1063.2 pg/cm2 at 1.0 N/m2, and 644.7 pg/cm2 at 1.5 N/m2. The average ET-1 secretion rate for the HGMEC monolayers exposed to 0.5, 1.0, or 1.5 N/m2 of shear stress was 32.83 +/- 2.01 pg/cm2 x h, 42.53 +/- 3.74 pg/cm2 x h, and 25.79 +/- 1.29 pg/cm2 x h, respectively. The average ET-1 secretion rate of the static controls was 28.22 +/ 3.11 pg/cm2 x h. The results showed that low shear stress (0.5 N/m2) elevated and high shear stress (1.5 N/m2) suppressed secretion of ET-1, while an intermediate level of shear stress (1.0 N/m2) led to the maximum secretion of ET 1, and furthermore, ET-1 secretion varied with the duration of shear in a nonlinear fashion, and the logistic equations may be used to describe relationship between the duration of shear and the ET-1 secretion. The major secretion period of ET-1 occurred between 5.3 and 22.3 h, with the peak secretion rate occurring at approximately 10.7-15.2 h. Our findings showed also that the major secretion period and peak secretion rate of HGMECs varied with the level of shear stress. Thus, the response of cultured human microvascular endothelial cells to shear stress differed from that of large-vessel endothelial cell cultures in terms of ET-1 secretion. In addition to the level of shear stress, the duration of shear is an important determinant in ET-1 secretion. Consequently, the heterogeneity of vascular endothelial cells and the duration of shear should both be considered in future research on the secretion of vascular endothelial cell cultures. PMID- 11866545 TI - Studies on single-cell adhesion probability between lymphocytes and endothelial cells with micropipette technique. AB - An in vitro model with micropipette technique was used to investigate single-cell adhesion probability between lymphocytes and endothelial cells. The basal adhesion probability between lymphocytes and endothelial cells was low and was significantly increased when either lymphocytes were activated by phytohemagglutinin (PHA) or endothelial cells were stimulated by tumor necrosis factor. The adhesion probability of lymphocytes to human umbilical vein endothelial cells was similar to that of lymphocytes to human brain microvascular endothelial cells (HB-MVEC). However, lymphocyte adhesion probability was higher in HB-MVEC than in mouse brain microvascular endothelial cells (MB-MVEC) under both resting and activated conditions. Furthermore, lymphocytes preincubated with monoclonal antibodies to lymphocyte function-associated antigen-1 (LFA-1) or HB MVEC preincubated with monoclonal antibodies to intercellular adhesion molecule 1 (ICAM-1) significantly down-regulated the adhesion probability between lymphocytes and endothelial cells, indicating that the adhesion probability is related to the expression of LFA-1 on lymphocytes and to the expression of ICAM-1 on endothelial cells. Lymphocytes isolated from patients with cerebral stroke exhibited increased adhesion probability to HB-MVEC as compared with lymphocytes from healthy donors. Preincubation of lymphocytes with tetramethylpyrazine (TMP), an extract from a Chinese traditional herb, effectively inhibited the adhesion probability to HB-MVEC, suggesting that TMP has a potential therapeutic value. These results indicate that the micropipette technique is a useful model for investigating single-cell adhesion probability between lymphocytes and endothelial cells in vitro. PMID- 11866546 TI - Inspiration-induced vascular responses in finger dorsum skin. AB - A rapid and deep inspiration triggers a sympathetically mediated transient vasoconstriction of skin arterioles (inspiratory gasp vascular response, IGVR). Because the IGVR has been most often measured and studied in skin that is rich in arteriovenous anastomoses (AVAs), such as the palmar aspect of the distal phalanx or plantar aspect of the toes, there is little information on its features in skin areas not dominated by thermoregulatory AVAs. Thus, the dependence of the magnitude of the IGVR on AVAs is unclear. We reasoned that if responses in a region of low AVA density, such as the finger dorsum distal phalanx, were comparable to those in AVA-rich skin, this would clarify the issue. Further, it might then be possible to use such areas to provide a useful complementary target for future study of sympathetically induced vasoconstriction. To test this, we determined the features of the finger dorsum IGVR in 28 healthy volunteers (age 19-57 years, 14 males) in whom distal phalanx skin blood perfusion (SBF) was monitored by laser-Doppler during 21 sequential IGVRs, each separated by 2 min. IGVR was quantified as the minimum SBF during each IGVR, expressed as a percentage of each immediately preceding 2-min SBF average. Results (mean +/- SD) revealed an overall IGVR of 72.2 +/- 16.7%, which is very near that reported from studies on the AVA-rich palmar finger pad. We therefore conclude that the IGVR does not depend on the presence of AVAs and that the dorsal distal phalanx is a viable alternative for the study of sympathetically related neurovascular responses. PMID- 11866547 TI - In vivo evidence for brain-to-blood efflux transport of valproic acid across the blood-brain barrier. PMID- 11866548 TI - Lipopolysaccharide-induced secretory phospholipase A2 activity in pericytes: a possible mechanism for mediating relaxation. PMID- 11866549 TI - Chemoselective protection of the amino groups of chitosan by controlled phthaloylation: facile preparation of a precursor useful for chemical modifications. AB - A simple and convenient procedure for chemoselectively protecting the amino groups of chitosan has been developed to provide N-phthaloyl-chitosan that is indispensable as a soluble N-protected precursor for further controlled modification reactions of chitosan. Although the conventional N-phthaloylation of chitosan in N,N-dimethylformamide was accompanied by partial phthaloylation of the hydroxy groups, the addition of a small amount of hydroxy-containing compounds effectively suppressed the O-phthaloylation. Of some compounds examined, water proved particularly suitable, resulting in the formation of chemoselectively N-phthaloylated chitosan without any appreciable O-phthaloyl groups. The resulting N-phthaloyl-chitosan was found to be crystalline despite the presence of a bulky substituent. A solubility test indicated that N-phthaloyl chitosan exhibited considerable affinity for organic solvents. PMID- 11866550 TI - The unique ribbon morphology of the major ampullate silk of spiders from the genus Loxosceles (recluse spiders). AB - The morphology of silk produced by recluse spiders (Loxosceles arizonica) was investigated by scanning electron microscopy, atomic force microscopy, and transmission electron microscopy. This silk consisted entirely of very long, thin ribbons of width 2-4 microm and thicknesses of no more than 40 nm. The correspondence in shape and dimension between the silk ribbons and the elongated aperture of the major ampullate spigot indicated that these ribbons were major ampullate silk. Selected area electron diffraction patterns from single ribbons were indexed with an orthorhombic unit cell (a = 9.43(2) A, b = 8.96(3) A, c = 6.96(1) A). This unit cell is in good agreement with that previously reported for synthetic poly(L-alanylglycine). Thus it is likely that the crystalline regions of the major ampullate silk of L. arizonica consist of an alternating glycine-L alanine motif that has adopted a beta-sheet structure. The amino acid composition achieved with the silk of L. arizonica as well as that of L. laeta confirmed that the major amino acid constituents of this silk were glycine and L-alanine in nearly equal amounts. As it was noticed that the dry ribbons were highly electrostatic, it is suggested that the electrostatic interaction plays an important role in prey capture for Loxoseles. PMID- 11866551 TI - A multitechnique approach in protein/surfactant interaction study: physicochemical aspects of sodium dodecyl sulfate in the presence of trypsin in aqueous medium. AB - Interaction of sodium dodecyl sulfate (SDS) with a globular protein, trypsin, has been physicochemically studied in aqueous medium in detail using tensiometric, conductometric, calorimetric, fluorimetric, viscometric, and circular dichroism techniques. The results indicate that SDS-trypsin aggregates start to form at a surfactant concentration higher than the critical micelle concentration of pure SDS micelle. In contrast, the counterion binding decreases in the presence of trypsin. The free energies and enthalpies of micellization, interfacial adsorption, and entropy of micellization associated with the interaction have also been calculated. The values show that the interaction phenomenon is entropy controlled and endothermic in nature. The increase in viscosity is observed for the system of SDS-trypsin cluster above the critical micelle concentration of SDS micelle only. The aggregation number and interface polarity decrease compared to the values of micelles without protein. Circular dichroism spectra show the high alpha-helical content and unfolded structure of trypsin in the presence of SDS due to strong electrostatic repulsion leading to a probable "necklace and bead" model in the case of biopolymer-surfactant complexes. PMID- 11866552 TI - The polymer-supported phospholipid bilayer: tethering as a new approach to substrate-membrane stabilization. AB - We present a new molecular engineering approach in which a polymer-supported phospholipid bilayer is vertically stabilized by controlled covalent tethering at both the polymer-substrate and polymer-bilayer interfaces. This approach is based on lipopolymer molecules, which not only form a polymer cushion between the phospholipid bilayer and a solid glass substrate but also act as covalent connections (tethers) between the bilayer and cushion. Our approach involves Langmuir-Blodgett transfer of a phospholipid-lipopolymer monolayer followed by Schaefer transfer of a pure phospholipid monolayer and is capable of varying the tethering density between the polymer layer and the phospholipid bilayer in a very controlled manner. Further stabilization is achieved if the glass substrate is surface-functionalized with a benzophenone silane. In this case, a photocross linking reaction between the polymer and benzophenone group allows for the covalent attachment of the polymer cushion to the glass substrate. This approach is similar to that recently reported by Wagner and Tamm in which double tethering is achieved via lipopolymer silanes (Wagner, M. L.; Tamm, L. K. Biophys. J. 2000, 79, 1400). To obtain a deeper understanding of how the covalent tethering affects the lateral mobility of the bilayer, we performed fluorescence recovery after photobleaching (FRAP) experiments on polymer-tethered bilayers at different tethering densities (lipopolymer/phospholipid molar ratios). The FRAP data clearly indicate that the hydrophobic lipopolymer moieties act as rather immobile obstacles within the phospholipid bilayer, thereby leading to hindered diffusion of phospholipids. Whereas the high lateral diffusion coefficient of D = 17.7 mum(2)/s measured at low tethering density (5 mol % lipopolymer) indicates rather unrestricted motion within the bilayer, corresponding values at moderate (10 mol % lipopolymer) and high (30 mol % lipopolymer) tethering densities of D = 9.7 mum(2)/s and D = 1.1 mum(2)/s, respectively, show significant hindered diffusion. These results are contrary to the recent findings on similar membrane systems reported by Wagner and Tamm in which no significant change in phospholipid diffusion was found between 0 and 10 mol % lipopolymer. Our experimental report leads to a deeper understanding of the complex problem of interlayer coupling and offers a path toward a compromise between stability of the whole system and lateral mobility within the bilayer. Furthermore, the FRAP measurements show that polymer-tethered membranes are very interesting model systems for studying problems of restricted diffusion within two-dimensional fluids. PMID- 11866553 TI - Study of blood compatibility with poly(2-methoxyethyl acrylate). Relationship between water structure and platelet compatibility in poly(2-methoxyethylacrylate co-2-hydroxyethylmethacrylate). AB - Previously, we reported that poly(2-methoxyethylacrylate) (PMEA) showed excellent blood compatibility and implied that the water structure in PMEA contributed to the blood compatibility. In this study, the relationship between the water structure and the blood compatibility is clarified by studying the influence of the monomer composition of poly(MEA-co-HEMA) on the water structure and the blood compatibility of the copolymers. The water in the polymer was classified into three types: free water, freezing bound water, and nonfreezing water. The polymers with 0-30 mol % of HEMA content had a significant amount of freezing bound water, and the amount decreases greatly when the composition of HEMA exceeded 30 mol %. On the other hand, the amount of other water increased simply with an increase of HEMA content. The evaluation of the platelet compatibility of poly(MEA-co-HEMA) revealed that the adhesion number and the morphological change of platelet on the copolymer surface were least when the HEMA content was 0-20 mol %. These two results strongly suggest that the freezing bound water relates to the platelet compatibility of the polymers. PMID- 11866554 TI - Mechanical characterization of an unusual elastic biomaterial from the egg capsules of marine snails (Busycon spp.). AB - Egg capsule material serves as a putative protection mechanism for developing snail embryos facing the perils of the marine environment. We conducted the first quantitative study of this acellular structural protein with the goals of characterizing its chemical and mechanical properties and the relationship of these properties to its biological protective function. We have found that this protein polymer exhibits long-range elasticity with an interesting recoverable yield evidenced by an order of magnitude decrease in elastic modulus (apparent failure) that begins at 3%-5% strain. This material differs significantly from other common structural proteins such as collagen and elastin in mechanical response to strain. Qualitative similarities in stress/strain behavior to keratin, another common structural protein, are more than coincidental when composition and detailed mechanical quantification are considered. This suggests the possibility of alpha-helical structure and matrix organization that might be similar in these two proteins. Indeed, the egg capsule protein may be closely related to vertebrate keratins such as intermediate filaments. We conclude that while this material's bimodal tensile properties may serve as useful protection against the impact loading egg capsules encounter in the intertidal zone, the full biological importance of these capsules is not known. PMID- 11866555 TI - Modulating fibroblast cell proliferation with functionalized poly(methyl methacrylate) based copolymers: chemical composition and monomer distribution effect. AB - Poly(methyl methacrylate)-based terpolymers bearing sulfonate and carboxylate groups have been synthesized by radical copolymerization leading to polymers with random distributions of ionic monomer units. Fibroblast cells were seeded on terpolymers of various molar compositions of ionic groups. Kinetics of the cell proliferation were examined and systematically compared to the nonfunctionalized control polymer, poly(methyl methacrylate). Modulation of cell proliferation was observed on 15% ionic monomer content copolymers of various compositions (R = COO(-)/(COO(-) + SO(3)(-)) and varies from 0 to 1). The inhibition percentage of cell proliferation calculated for each polymer by comparison to the cell proliferation on the control was plotted against R and gave a maximum value for R close to 0.55. Copolymers with ionic group contents higher or lower than 15% exhibit inhibition percentages fitting with those previously observed for the same R values, showing that the hydrophilic properties are not sufficient to explain the modulation effect of this material toward cells. Moreover, for each polymer tested, cells, even if inhibited in growth, were shown to be viable, indicating that the synthesized terpolymers exhibit cytostatic properties excluding any cytotoxic effect. Such polymers may be used for the fabrication of biocompatible intraocular lenses and prevent secondary cataract. PMID- 11866556 TI - Oxypropylation of cork and the use of the ensuing polyols in polyurethane formulations. AB - Cork particles, recovered as byproducts of the processing of this natural material, were oxypropylated under pressure and relatively high temperature in the presence of KOH as catalyst. Various parameters were explored in order to assess the most suitable conditions, which led to the almost complete conversion of the solid cork into a viscous polyol. This product was a mixture of oxypropylated cork macromolecules and propylene oxide oligomers, which were thoroughly characterized. The use of these polyols as macromonomers in the synthesis of polyurethane foams gave promising results, thus showing that it should be possible to exploit the residues of this important renewable resource to manufacture original materials. PMID- 11866557 TI - Biomimetic poly(methyl methacrylate)-based terpolymers: modulation of bacterial adhesion effect. AB - Adherence of Staphylococcus aureus, responsible for major foreign body infections, was assessed onto functionalized poly(methyl methacrylate)-based terpolymers bearing sulfonate and carboxylate groups and onto poly(methyl methacrylate) as control. These terpolymers, have been synthesized by radical copolymerization of methyl methacrylate, methacrylic acid, and sodium styrene sulfonate by varying the ratio R = [COO(-)]/[COO(-) + SO(3)(-)] from 0 to 1 and keeping ionic monomer content between 7 and 18%. Adsorption of fibronectin onto poly(methyl methacrylate) was shown to dramatically promote bacterial adherence, whereas a strong inhibition of bacteria adherence was observed onto functionalized terpolymers containing both carboxylate and sulfonate groups. When terpolymers were predominantly functionalized by carboxylate groups, bacteria adherence was favored and reached values close to those obtained for poly(methyl methacrylate). These results have been related to the distribution of the anionic groups along the macromolecular chains, creating active sites responsible for specific interactions with fibronectin and inducing modifications of its conformation. The conformation of the adsorbed adhesive protein was then suggested to have an influence on the availability of its interaction sites to bacteria adhesins and therefore on modulation of bacteria adherence. Inhibition of Staphylococcus aureus adherence by functionalized poly(methyl methacrylate) based terpolymers is of great interest in the field of biomedical implants and especially in the case of ophthalmic applications. PMID- 11866558 TI - Comparison of metal-ion-dependent cleavages of RNA by a DNA enzyme and a hammerhead ribozyme. AB - Joyce's DNA enzyme catalyzes cleavage of RNAs with almost the same efficiency as the hammerhead ribozyme. The cleavage activity of the DNA enzyme was pH dependent, and the logarithm of the cleavage rate increased linearly with pH from pH 6 to pH 9 with a slope of approximately unity. The existence of an apparent solvent isotope effect, with cleavage of RNA by the DNA enzyme in H(2)O being 4.3 times faster than cleavage in D(2)O, was in accord with the interpretation that, at a given pH, the concentration of the active species (deprotonated species) is 4.3 times higher in H(2)O than the concentration in D(2)O. This leads to the intrinsic isotope effect of unity, demonstrating that no proton transfer occurs in the transition state in reactions catalyzed by the DNA enzyme. Addition of La(3+) ions to the Mg(2+)-background reaction mixture inhibited the DNA enzyme catalyzed reactions, suggesting the replacement of catalytically and/or structurally important Mg(2+) ions by La(3+) ions. Similar kinetic features of DNA enzyme mediated cleavage of RNA and of hammerhead ribozyme-mediated cleavage suggest that a very similar catalytic mechanism is used by the two types of enzyme, despite their different compositions. PMID- 11866559 TI - Crystallinity and morphology in films of starch, amylose and amylopectin blends. AB - Films of potato starch, amylose, and amylopectin and blends thereof were prepared by solution casting and examined using X-ray diffraction, light microscopy, transmission electron microscopy, and differential scanning calorimetry. Amylose films had a relative crystallinity of about 30% whereas amylopectin films were entirely amorphous. Blending of amylose and amylopectin resulted in films with a considerably higher degree of crystallinity than could be predicted. This is explained by cocrystallization between amylose and amylopectin and possibly by crystallization of amylopectin. The crystallized material gave rise to an endotherm detected with differential scanning calorimetry. The enthalpy and peak temperature of the transition also increased as the water content decreased. When the amylose proportion in the blends was low, separate phases of amylose and amylopectin were observed by light microscopy. At higher amylose proportions, however, the phase separation was apparently prevented by amylose gelation and the formation of a continuous amylose network. The amylose network in the films, observed with transmission electron microscopy, consisted of stiff strands and open pores and became less visible as the amylose proportion decreased. The water content of the films was dependent on the microstructure and the crystallinity. PMID- 11866560 TI - Microheterogeneity and microrheology of wheat gliadin suspensions studied by multiple-particle tracking. AB - By monitoring the thermally driven displacements of imbedded polystyrene microspheres via video fluorescence microscopy, we quantified the microstructural and micromechanical heterogeneities of wheat gliadin suspensions. We found that the degree of heterogeneity of the suspensions, as measured by the width and skewness of the microspheres' mean squared displacement (MSD) distribution, increased dramatically over a narrow range of gliadin concentrations. The ensemble-averaged MSD of a 250 mg/mL gliadin suspension exhibited a power-law behavior scaling linearly with time, a behavior similar to that observed for a homogeneous aqueous glycerol solution. However, the MSD distribution was wider and more asymmetric than for glycerol. With increasing concentration of gliadin, the ensemble-averaged MSD rapidly displayed a plateau at small time scales, the MSD distribution became wider and more asymmetric, and the local viscoelastic moduli extracted from multiple-particle-tracking measurements showed an increasingly wide range. PMID- 11866562 TI - Biological thermal detection: micromechanical and microthermal properties of biological infrared receptors. AB - Bioinspired design of biomimetic sensors relies upon the complete understanding of properties and functioning of biological analogues in conjunction with an understanding of their microstructural organization at various length scales. In the spirit of this approach, the microscopic properties of infrared (IR) receptors of snakes with "infrared vision" were studied with scanning thermal microscopy and micromechanical analysis. Low surface thermal conductivity of 0.11 W/(m K) was measured for the IR receptor surfaces as compared to the nonspecific skin areas. This difference in surface thermal conductivity should result in a significant local temperature gradient around the receptor areas. Micromechanical analysis showed that pit organs were more compliant than surrounding skin areas with an elastic modulus close to 40 MPa. In addition, the maximum elastic modulus was detected for the outermost layer with gradually reduced elastic resistance for the interior. The porous microstructure of the underlying tissue combined with the highly branched microfibrillar network (Biomacromolecules 2001, 2, 757) is thought to be responsible for such a combination of biomaterial properties. Considering these biomaterials features, we postulated a possible design of an artificial photothermal detector inspired by the microstructure of natural receptors. This bioinspired design would include a microfabricated cavity filled with an ordered lattice of microspheres with a gradient periodicity from the surface to the interior. Such a "photonic cavity" could provide an opportunity for multiple scattering at wavelength tuned to 8-12 microm as a range of highest sensitivity. PMID- 11866561 TI - Characterization of the spontaneously forming hydrogels composed of water-soluble phospholipid polymers. AB - Spontaneously forming hydrogels composed of 2-methacryloyloxyethyl phosphorylcholine (MPC) copolymers, poly(MPC-co-methacrylic acid) (PMA), and poly(MPC-co-n-butyl methacrylate) (PMB) were examined. The MPC copolymer hydrogel was observed to have a spontaneous gelation property. To determine the properties of the hydrogels and why the gelation takes place, we have studied the properties of the hydrogels by scanning electron microscopy, X-ray photoelectron spectroscopy (XPS), and differential scanning calorimetry (DSC). The morphologies of the hydrogels were spongelike with a homogeneous structure. By XPS analysis in terms of the molecular distributions in the hydrogels, it was observed that a stabilization time was required for the hydrogel to undergo chain rearrangement. DSC thermograms of the hydrogels were different from their components, PMA and PMB. For the hydrogel, a crystallization peak around -30 degrees C was observed. This result indicated that some ordered structures existed in the hydrogels. To determine the role of the MPC groups, aqueous solutions of poly(methacrylic acid) (PMAc) and PMB were mixed. The mixture of PMAc-PMB turned into a sol state, and the sol state remained for a week. When the mixture was cooled, a very weak hydrogel was prepared. This result suggested that the MPC groups were the dominant unit for spontaneously forming the hydrogels. PMID- 11866563 TI - Amphiphilic block copolymers as bile acid sorbents: 1. Synthesis of polystyrene-b poly(N,N,N-trimethylammoniumethylene acrylamide chloride). AB - The systematic investigation of the synthesis of polystyrene-b-poly(N,N,N trimethylammoniumethylene acrylamide chloride) was accomplished by employing both polystyrene-b-poly(tert-butyl acrylate) and its hydrolyzed derivative, polystyrene-b-poly(acrylic acid) (PS-b-PAA) as starting materials, and coupling them with N,N-dimethylethylenediamine (DMED). The various reactions and intermediates we examined include aluminum amides, acid chlorides, and imides derived from carbodiimides, all in a variety of solvents. We present below our investigation of several synthetic routes and conclude that the carbodiimide coupling of PS-b-PAA with DMED followed by quaternization and counterion exchange is the most effective method of achieving the target. A brief discussion of the merits of each procedure in the context of block copolymers is given, and IR spectroscopic evidence for the postpolymerization synthesis of the poly(acrylamide) block is provided. PMID- 11866564 TI - Amphiphilic block copolymers as bile acid sorbents: 2. Polystyrene-b-poly(N,N,N trimethylammoniumethylene acrylamide chloride): self-assembly and application to serum cholesterol reduction. AB - This paper presents morphological studies and preliminary bile salt binding properties of the new amphiphilic diblock copolymer polystyrene-b-poly(N,N,N trimethylammoniumethylene acrylamide chloride) (PS-b-PTMEACl)(1) (see Figure 1), a derivative of PS-b-poly(tert-butylacrylate) (PS-b-PtBuA). In an aqueous environment, PS-b-PTMEACl forms simple spheres (approximately 20 nm diameter), large compound micelles (>100 nm diameter), and larger, more complex architectures as presented and discussed below. The colloidal stability with respect to sodium chloride and as a function of particle concentration is also considered. Finally, PS-b-PTMEACl aggregates were prepared and tested as an alternative to the commercially available bile salt sequestrant resins that target coronary heart disease due to elevated cholesterol levels. Electron micrographs were employed to visualize the colloid-based polyelectrolyteminus signbiosurfactant interaction and chromatographic separation analytical methods were used to quantify the sequestration. The results indicate that although at this preliminary stage they require laborious preparation, self-assembled aggregates may present an interesting alternative to the clinically used bile salt sequestrants. PMID- 11866565 TI - Side-chain effect of second monomer units on crystalline morphology, thermal properties, and enzymatic degradability for random copolyesters of (R)-3 hydroxybutyric acid with (R)-3-hydroxyalkanoic acids. AB - Three types of random copolymers with 94 mol % (R)-3-hydroxybutyric acid (3HB) and 6 mol % (R)-3-hydroxyalkanoic acids with different side-chain lengths, (R)-3 hydroxypentanoic acid (3HV), (R)-3-hydroxyhexanoic acid (3HHx), and medium-chain length (R)-3-hydroxyalkanoic acids (mcl-3HA, C8-C12), were prepared by biological synthetic techniques. The solid-state structure and thermal properties of melt crystallized films for copolymers were characterized by means of wide-angle X-ray diffraction, small-angle X-ray scattering, differential scanning calorimetry, and optical microscopy. The randomly distributed second monomer units, except for 3HV in copolyesters, act as defects of the P(3HB) crystal and are excluded from the P(3HB) crystalline lamellae. The lamellar thickness of copolymers decreased with an increase in the side-chain length of second monomer units. In addition, the growth rate of spherulites decreased with an increase in the carbon numbers of second monomer units at an identical crystallization temperature. These results indicate that a steric bulkiness of the second monomer unit affects the crystallization of (R)-3HB segments in random copolyesters. An enzymatic degradation test of melt-crystallized copolymer films was carried out in the presence of PHB depolymerase from Alcaligenes faecalis T1. Erosion rate of copolyesters was dependent on both the crystallinity and the lamellar thickness of samples. As the result, the rate of enzymatic degradation for copolymer films increased with an increase in the carbon numbers of second monomer units. PMID- 11866567 TI - Graft copolymerization of methyl methacrylate onto mercaptochitin and some properties of the resulting hybrid materials. AB - The graft copolymerization of methyl methacrylate onto mercaptochitin and some properties of the resulting graft copolymers have been studied. Methyl methacrylate was efficiently graft copolymerized onto mercaptochitin in dimethyl sulfoxide, and the grafting percentage reached 1300% under appropriate conditions. Although the side-chain ester groups were resistant to aqueous alkali, hydrolysis could be achieved with a mixture of aqueous sodium hydroxide and dimethyl sulfoxide. Subsequent treatment with acetic anhydride in methanol transformed the sodium carboxylate groups into carboxyl groups. Although the graft copolymers exhibited an improved affinity for organic solvents, those having sodium carboxylate or carboxyl units were characterized by a much more enhanced solubility and were soluble in common solvents. The hygroscopic nature of chitin decreased with an increase in the grafting extent but increased significantly upon hydrolysis of the ester groups. The enzymatic degradability of the graft copolymers, as evaluated with lysozyme, was also dependent on the grafting extent and much higher than that of the original chitin. DSC measurements revealed the presence of a glass transition phenomenon, which could be ascribed to the poly(methyl methacrylate) side chain. PMID- 11866566 TI - Assembly of electroactive layer-by-layer films of hemoglobin and polycationic poly(diallyldimethylammonium). AB - Layer-by-layer (PDDA/Hb)(n) films were assembled by alternate adsorption of positively charged poly(diallyldimethylammonium) (PDDA) and negatively charged hemoglobin (Hb) at pH 9.2 from their aqueous solutions on pyrolytic graphite electrodes and other substrates. The assembly process was monitored and confirmed by quartz crystal microbalance (QCM), UV-vis spectroscopy, and cyclic voltammetry (CV). CVs of (PDDA/Hb)(n) films showed a pair of well-defined, nearly reversible peaks at about -0.34 V vs SCE at pH 7.0, characteristic of Hb heme Fe(III)/Fe(II) redox couple. Positions of Soret absorption band and infrared amide II band of Hb in (PDDA/Hb)(8) films suggest that Hb in the films keeps its secondary structure similar to its native state. The electrochemical parameters of (PDDA/Hb)(8) films were estimated by square wave voltammetry, and the thickness of the PDDA/Hb bilayer was estimated by QCM and scanning electron microscopy. Trichloroacetic acid and nitrite (NO(2)(-)) were catalytically reduced at (PDDA/Hb)(8) film electrodes. The electrochemical catalytic reactions of O(2) and H(2)O(2) on (PDDA/Hb)(8) films were also studied. PMID- 11866568 TI - Synthesis and characterization of triblock copolymers of methoxy poly(ethylene glycol) and poly(propylene fumarate). AB - Amphiphilic block copolymers were synthesized by transesterification of hydrophilic methoxy poly(ethylene glycol) (mPEG) and hydrophobic poly(propylene fumarate) (PPF) and characterized. Four block copolymers were synthesized with a 2:1 mPEG:PPF molar ratio and mPEGs of molecular weights 570, 800, 1960, and 5190 and PPF of molecular weight 1570 as determined by NMR. The copolymers synthesized with mPEG of molecular weights 570 and 800 had 1.9 and 1.8 mPEG blocks per copolymer, respectively, as measured by NMR, representing an ABA-type block copolymer. The number of mPEG blocks of the copolymer decreased with increasing mPEG block length to as low as 1.5 mPEG blocks for copolymer synthesized with mPEG of molecular weight 5190. At a concentration range of 5-25 wt % in phosphate buffered saline, copolymers synthesized with mPEG molecular weights of 570 and 800 possessed lower critical solution temperatures (LCST) between 40 and 45 degrees C and between 55 and 60 degrees C, respectively. Aqueous solutions of copolymer synthesized with mPEG 570 and 800 also experienced thermoreversible gelation. The sol-gel transition temperature was dependent on the sodium chloride concentration as well as the mPEG block length. The copolymer synthesized from mPEG 570 had a transition temperature between 40 and 20 degrees C with salt concentrations between 1 and 10 wt %, while the sol-gel transition temperatures of the copolymer synthesized from mPEG molecular weight 800 were higher in the range 75-30 degrees C with salt concentrations between 1 and 15 wt %. These novel thermoreversible copolymers are the first biodegradable copolymers with unsaturated double bonds along their macromolecular chain that can undergo both physical and chemical gelation and hold great promise for drug delivery and tissue engineering applications. PMID- 11866569 TI - Characterization of microbial polythioesters: physical properties of novel copolymers synthesized by Ralstonia eutropha. AB - Various samples of polythioesters with different contents of 3-mercaptopropionic acid (3MP) or 3-mercaptobutyric acid (3MB) as one comonomer and with 3 hydroxybutyric acid (3HB) as the second constituent were produced by cultivating cells of Ralstonia eutropha strain H16 in mineral salts medium containing 3MP or 3MB plus gluconate as carbon sources. Fermentations were done also at the 30-L scale. The various samples were cast as films from chloroform and the following were recorded: melting point, solid-state NMR, X-ray diffraction. The copolyester poly(3HB-co-3MP) displayed mutiple melting peaks corresponding to separate phases rich in 3MP and 3HB. The copolyester poly(3HB-co-3MB) displayed very low crystallinity and melting points higher than that of poly(3HB) when the 3HB content was 40% or less. PMID- 11866570 TI - Cell membrane as a model for the design of ion-active nanostructured supramolecular systems. AB - The synthesis and polymerization of six AB(3) tapered self-assembling methacrylate monomers (5a, 5b, 5c,5d, 17a, and 17b) based on first generation alkyl substituted benzyl ether monodendrons (i.e., minidendrons) containing oligooxyethylene units at their focal point and the polymerizable group on their periphery are described. The corresponding polymers (6a, 6b, 6c, 6d, 18a, and 18b) self-assemble and subsequently self-organize in supramolecular networks that form a 2-D hexagonal lattice. This network consists of a continuous phase based on a paraffin barrier material perforated in a hexagonal array by ion-active channels constructed from the oligooxyethylenic units protected by the aromatic groups of the taper. Complexation of the oligooxyethylene channels of 6a-d with LiCF(3)SO(3) salt enhances the thermal stability of their hexagonal columnar (phi(h)) liquid crystalline phase. The enhancement of the thermal stability of the phi(h) phase of both monomers and polymers up to 86 degrees C is also achieved by shifting the placement of the polymerizable group from the 3 position to the 4 position of the 3,4,5-trisubstituted AB(3) benzoate monodendrons. The design of these macromolecules was inspired by the bilayer fluid mosaic structure of the cell membrane. The lipid bilayer of the cell membrane that acts in its ordered state as a barrier to the passage of polar molecules was replaced with the paraffinic barrier, while the protein-based ionic channels were replaced with oligooxyethylenic-based channels. The resulted supramolecular material has the mechanical integrity required for the design of ion-active nanostructured supramolecular systems. PMID- 11866571 TI - X-ray microbeam and electron diffraction experiments on developing xylem cell walls. AB - This paper describes the first successful application of the novel technique of X ray microbeam diffraction to the study of wood cell walls in developing xylem tissue. The method enabled us to obtain quantitative diffraction diagrams from single cell walls. It was further combined with selected area electron diffraction. We find that the crystal size of cellulose is increasing from the primary wall (P, <19 A) over the outer layer (S(1), 19 A) to the middle layer of the secondary wall (S(2), 24 A). In particular, the cellulose crystals formed in the primary wall consist of an extremely low-crystalline state of cellulose I. We suggest the more applicable concept of microfibrils with increasing lateral disorder, similar to cellulose IV(I). PMID- 11866572 TI - Peroxidase-catalyzed oxidative polymerization of bisphenols. AB - Oxidative polymerization of bisphenolic monomers has been performed using peroxidase as catalyst in an aqueous organic solvent. Peroxidase induced the polymerization of an industrial product, bisphenol F, consisting of 2,2'-, 2,4'-, and 4,4'-dihydroxydiphenylmethanes. Under the selected conditions, the quantitative formation of the polymer was observed. Among the isomers, 2,4'- and 4,4'-dihydroxydiphenylmethanes were polymerized to give the polymer in high yields, whereas no polymerization of the 2,2'-isomer occurred. These data suggest that the radical transfer reaction between a phenoxy radical of the enzymatically polymerizable monomer and the enzymatically nonpolymerizable monomer frequently took place during the polymerization. Various 4,4'-dihydroxyphenyl compounds were also polymerized by peroxidase catalyst. The polymerization behaviors, and solubility and thermal properties of the resulting polymers strongly depended on the bridge structure as well as the enzyme origin. Polymers from dihydroxydiphenylmethanes showed relatively high thermal stability. PMID- 11866573 TI - Amine-reactive biodegradable diblock copolymers. AB - A new class of diblock copolymers was synthesized from biodegradable poly(lactic acid) and poly(ethylene glycol)minus signmonoamine. These polymers were activated by covalently attaching linkers such as disuccinimidyl tartrate or disuccinimidyl succinate to the hydrophilic polymer chain. The polymers were characterized by (1)H NMR spectroscopy, (13)C NMR spectroscopy and gel permeation chromatography (GPC). These investigations indicated that the polymers were obtained with the correct composition, in high purities, and the expected molecular weight. By using dyes containing primary amine groups such as 5-aminoeosin as model substrates, it was possible to show that the polymers are able to bind such compounds covalently. The diblock copolymers were developed to suppress unspecific protein adsorption and allow the binding of bioactive molecules by instant surface modification. The polymers are intended to be used for tissue engineering applications where surface immobilized cell adhesion peptides or growth factors are needed to control cell behavior. PMID- 11866574 TI - Formation of and coalescence from the inclusion complex of a biodegradable block copolymer and alpha-cyclodextrin. 2: A novel way to regulate the biodegradation behavior of biodegradable block copolymers. AB - A biodegradable block copolymer (PCL-b-PLLA, M(n) = 1.72 x 10(4), M(w)/M(n) = 1.37) of poly(epsilon-caprolactone) (PCL) and poly(L-lactide) (PLLA) with very low crystallinity was obtained by forming the inclusion complex between alpha cyclodextrin molecules and PCL-b-PLLA followed by coalescence of the guest polymer chains. Films of the as-synthesized and coalesced copolymer samples, PCL and PLLA homopolymers of approximately the same chain lengths as the corresponding blocks of PCL-b-PLLA, and a physical blend of PCL/PLLA homopolymers with the same molar composition as PCL-b-PLLA were prepared by melt-compression molding between Teflon plates. Subsequently, the in vitro biodegradation behavior of these films was studied in phosphate buffer solution containing lipase from Rhizopus arrhizus, by means of ultraviolet spectra, attenuated total reflectance FTIR spectra, differential scanning calorimetry, wide-angle X-ray diffraction measurements, and weight loss analysis. PCL segments were found to degrade much faster than PLLA segments, both in the pure state and in copolymer or blend samples. Consistent with our expectation, suppression of the phase separation, as well as a decrease of crystallinity, in the coalesced copolymer sample led to a much faster enzymatic degradation than that of either as-synthesized copolymer or the PCL/PLLA physical blend sample, especially during the early stages of biodegradation. Thus the biodegradation behavior of biodegradable block copolymers, which is of decisive importance in drug delivery and controlled release systems, may be regulated by the novel and convenient means recently reported by us.(1) PMID- 11866575 TI - Formation of short chain length/medium chain length polyhydroxyalkanoate copolymers by fatty acid beta-oxidation inhibited Ralstonia eutropha. AB - Ralstonia eutropha has been considered as a bacterium, incorporating hydroxyalkanoates of less than six carbons only into polyhydroxyalkanoates (PHAs). Cells of the wild type cultivated with sodium octanoate as the carbon source in the presence of the fatty acid beta-oxidation inhibitor sodium acrylate synthesized PHAs composed of the medium chain length hydroxyalkanoates (3HA(MCL)) 3-hydroxyhexanoate (3HHx) and 3-hydroxyoctanoate (3HO) as well as of 3 hydroxybutyrate and 3-hydroxyproprionate as revealed by gas chromatography, (1)H NMR spectroscopy, and mass spectroscopy. The characterization of the polymer as a tetrapolymer was confirmed by differential solvent extraction and measurement of melting and glass transition temperature depression in the purified polymer compared to PHB. These data suggested that the R. eutropha PHA synthase is capable of incorporating longer chain substrates than suggested by previous in vitro studies. Furthermore, expression of the class II PHA synthase gene phaC1 from P. aeruginosa in R. eutropha resulted in the accumulation of PHAs consisting of 3HA(MCL) contributing about 3-5% to cellular dry weight. These PHAs were composed of nearly equal molar fractions of 3HO and 3-hydroxydecanoate (3HD) with traces of 3HHx. These data indicated that 3HA(MCL)-CoA thioesters were diverted from the fatty acid beta-oxidation pathway towards PHA biosynthesis in recombinant R. eutropha. PMID- 11866577 TI - Thermal degradation of poly(3-hydroxyalkanoates): preparation of well-defined oligomers. AB - The thermal degradation of the biodegradable bacterial polyesters poly(3 hydroxybutyrate), PHB, poly(3-hydroxyvalerate), PHV, and poly(3-hydroxybutyrate co-3-hydroxyvalerate), 0-21 mol % of hydroxyvalerate, was studied. At moderately low temperatures (170-200 degrees C), the main product is a well-defined oligomer, especially a 500-10,000 g/mol macromolecule, which contains one unsaturated end group, predominantly a trans-alkenyl end group, as well as a carboxylic end group. The process was studied regarding the effect of the copolymer composition and reaction time at 190 degrees C. During the first few hours of reaction, the thermal degradation of PHB and PHV followed a kinetic model of random scission, but eventually auto-acceleration of the pyrolysis was detected, probably due to the influence of the crotonate end groups of the oligomers formed. Ten-time scale-up experiments on a Brabender instrument were successfully undertaken. PMID- 11866576 TI - NMR imaging of high-amylose starch tablets. 1. Swelling and water uptake. AB - Pharmaceutical tablets made of modified high-amylose starch have a hydrophilic polymer matrix into which water can penetrate with time to form a hydrogel. Nuclear magnetic resonance imaging was used to study the water penetration and the swelling of the matrix of these tablets. The tablets immersed in water were imaged at different time intervals on a 300 MHz NMR spectrometer. Radial images show clearly the swelling of the tablets and the water concentration profile. The rate constants for water diffusion and the tablet swelling were extracted from the experimental data. The water diffusion process was found to follow case II kinetics at 25 degrees C. NMR imaging also provided spin density profiles of the water penetrating inside the tablets. PMID- 11866578 TI - Calculations of the contribution of ring currents to the chemical shielding anisotropy. AB - Ring currents can exert a large effect upon the chemical shielding of NMR resonances. The analytical expression developed by Waugh and Fessenden (Waugh, J. S.; Fessenden, R. W. J. Am. Chem. Soc. 1957, 79, 846) and Johnson and Bovey (Johnson, C. E.; Bovey, F. A. J. Chem. Phys. 1957, 29, 1012) only quantifies the contribution of ring currents to the isotropic component of the shielding tensor. In the work described here an additional analytical expression is developed so that the contribution of ring currents to the full shielding tensor can be calculated, allowing an estimate of the influence of ring currents upon the chemical shielding anisotropy (CSA, Deltasigma). To test that this pair of analytical expressions can provide a reasonable estimate of the contribution of ring currents to the full shielding tensor a series of density functional calculations (DFT) were carried out. A shielding tensor in a model compound was calculated in two distinct ways. For the first series, DFT shielding calculations of the model compound were carried out in the presence of a benzene ring. For the second series a ring current contribution to the shielding tensor was calculated via the new expressions, and this was added to the result of a DFT shielding calculation which used in place of benzene the nonaromatic analogue 1,3 cyclohexadiene. The two series of results proved to be in excellent agreement. The pair of analytical expressions are used to calculate ring current contributions to the CSA (Deltasigma) of 1H(N) backbone amide resonances in a structure of the second type 2 module from the protein fibronectin. Significant CSA variations are predicted in particular for the 1H(N) of G42 which is most likely involved in a N-H...tpi aromatic hydrogen bond. PMID- 11866579 TI - New phosphate fluorosurfactants for carbon dioxide. AB - Anionic phosphate fluorosurfactants were shown to self-assemble into water-in carbon dioxide microemulsions. The surfactants, having either two fluorinated chains or one fluorinated chain and one hydrocarbon chain, facilitated significant water uptake in CO2. Small angle neutron scattering (SANS) measurements of surfactant/water/CO2 solutions confirmed the presence of nanometer-scale aggregates, indicative of microemulsion formation. PMID- 11866580 TI - Adding L-3-(2-Naphthyl)alanine to the genetic code of E. coli. AB - An unnatural amino acid, L-3-(2-naphthyl)alanine, has been site-specifically incorporated into proteins in Escherichia coli. An orthogonal aminoacyl-tRNA synthetase was evolved that uniquely aminoacylates the unnatural amino acid onto an orthogonal amber suppressor tRNA, which delivers the acylated amino acid in response to an amber nonsense codon with translational fidelity greater than 99%. This result, together with the in vivo site-specific incorporation of O-methyl-L tyrosine reported previously, demonstrate that this methodology may be applicable to a host of amino acids. The expansion of the genetic code to include amino acids beyond the common 20 would provide an opportunity to better understand and possibly enhance protein (and perhaps organismal) function. PMID- 11866581 TI - Toward synthetic transcription activators: recruitment of transcription factors to DNA by a PNA-peptide chimera. AB - A PNA-peptide chimera designed to mimic the biochemical function of transcription activators has been synthesized and characterized. The bis-PNA segment binds specifically to a DNA site while the 20-residue peptide is capable of binding to the transcription factors Gal11 and Gal80. The PNA-peptide chimera thus mimics one of the central functions of a native transcription activator, recruitment of transcription factors to a specific DNA site. PMID- 11866582 TI - The effect of varied ion distributions on long-range DNA charge transport. AB - Long-range oxidative damage to DNA was utilized as a probe to delineate the effects of different ion distributions on DNA charge transport. DNA assemblies were constructed, containing a tethered rhodium intercalating photooxidant, spatially separated from two 5'-GG-3' sites of oxidative damage, with either an A6-tract or a mixed DNA sequence intervening between the guanine doublets; the extent of charge transport was assessed through measurements of the ratio of yields of damage at the guanine doublet distal versus that proximal to the metal binding site. The distal/proximal damage ratios were compared after photooxidation of otherwise identical Rh-tethered assemblies, except for 32P labeling either at the 5'- or 3'-end; this labeling difference corresponds, in the absence of charge neutralization by condensed counterions, to a shift in negative charge from one end of the duplex to the other. Both with assemblies containing the mixed sequence and the A6-tract, we observed that moving the negative charges to the proximal end of the duplex significantly decreased hole transport to the distal end. We propose that these results reflect variations in the thermodynamic potential at the proximal and distal guanine sites because of the change in charges at the termini of the oligomer. High values for the internal dielectric constant of the stacked base pairs are suggested by these data. Hence, the longitudinal polarizability of DNA may be important to consider in mechanisms for long-range DNA charge transport. PMID- 11866583 TI - A highly enantioselective amino acid-catalyzed route to functionalized alpha amino acids. AB - The development of syntheses providing enantiomerically pure alpha-amino acids has intrigued generations of chemists and been the subject of intense research. This report describes a general approach to functionalized alpha-amino acids based on catalytic asymmetric synthesis. Proline catalyzed Mannich-type reactions of N-PMP-protected alpha-imino ethyl glyoxylate with a variety of unmodified ketones to provide functionalized alpha-amino acids in high yields with excellent regio-, diastereo-, and enantioselectivities. Study of seven examples yielded six with product ee values of > or = 99%. In reactions involving ketone donors where diastereoisomeric products could be formed, two adjacent stereogenic centers were created simultaneously upon carbon-carbon bond formation with complete syn stereocontrol. Significantly, this methodology utilizes readily available and rather inexpensive starting materials, does not require any preactivation of substrates or metal ion assistance, and can be carried out on a gram scale under operationally simple reaction conditions. The keto-functionality present in the products provides a particularly attractive site for versatile modifications. This study compliments and extends our bioorganic approach to asymmetric synthesis to a versatile synthon class. Given that we have shown that a variety of optically active amino acids can be synthesized with proline catalysis, where an L-amino acid begets other L-amino acids, our results may stimulate thoughts concerning prebiotic syntheses of optically active amino acids based on this route. PMID- 11866584 TI - Microsomal P450-catalyzed N-dealkylation of N,N-dialkylanilines: evidence for a C(alpha)-H abstraction mechanism. AB - The early proposal that P450-catalyzed N-dealkylation of N,N-dialkylamines proceeds through a single-electron-transfer (SET) mechanism was later challenged in favor of the C(alpha)-H abstraction mechanism. In the present study, a series of N-alkyl-N-cyclopropyl-p-chloroaniline probes have been used to examine whether the P450-catalyzed N-dealkylations proceed through a C(alpha)-H abstraction and/or a SET mechanism, using phenobarbital-induced rat liver microsomal P450 enzymes as a model system. While the findings are highly consistent with a C(alpha)-H abstraction mechanism, further experimental evidence may be necessary to completely rule out the SET mechanism. PMID- 11866585 TI - Protein dynamics and cytochrome c: correlations between ligand vibrations and redox activity. AB - A recently developed method to directly observe specific protein vibrations, based on deuteration, has been employed to examine the redox-dependent structural and fluctional properties of cytochrome c. The dynamics of the protein-based methionine heme ligand were examined by selectively deuterating the ligand's methyl group. The frequency and line width of the C-D bonds were easily observable and shown to be sensitive to mutation-induced changes in the protein redox potential. However, of seven mutants examined, the C-D line widths were independent of the redox-state of the protein. Therefore, although the ligand dynamics depend on the protein's redox state, there are no detected differences in protein dynamics of the oxidized and reduced proteins. PMID- 11866586 TI - SCMTR: a chloride-selective, membrane-anchored peptide channel that exhibits voltage gating. AB - The design, synthesis, and characterization of a synthetic chloride membrane transporter, SCMTR (synthetic chloride membrane transporter, "scimitar"), are presented. The compound [CH3(CH2)17]2NCOCH2OCH2CO-GGGPGGG-OBzl inserts rapidly into liposomes and planar lipid bilayers. SCMTR has a 1.3 +/- 0.01 nS chloride diffusion pathway (>10:1 Cl/K selectivity). Evidence was also obtained for voltage gating behavior. PMID- 11866587 TI - Biocatalytic route to well-defined macromers built around a sugar core. AB - By using 4-C-hydroxymethyl-alpha-D-pentofuranose as the sugar core and lipase catalyzed transformations, a macromer was constructed with exceptional control of substituent placement around the carbohydrate core. The key synthetic transformations performed were as follows: (1) selective lipase-catalyzed acrylation along with prochiral selection of 4-C-hydroxymethyl-1,2-O isopropylidene-alpha-D-pentofuranose (diastereomeric excess up to 93%); (2) the ring-opening of epsilon-caprolactone, epsilon-CL, from the remaining primary hydroxyl group to give an acryl-sugar capped macromer (M(n) = 11 300, M(w)/M(n) = 1.36, initiator efficiency 50-55%, <5% water initiated PCL chains); (3) selective lipase-catalyzed esterification of the terminal hydroxyl of oligo(epsilon-CL) chains; (4) hydrolysis of the 1,2-O-isopropylidene group at the sugar core without any substantial loss in macromer molecular weight; and (5) homopolymerization of the corresponding macromer. In principle, the method developed is flexible so that it can be used to generate a wide array of unusual macromers and heteroarm stars. In the absence of biocatalytic transformation, such structural control would be extremely difficult or currently impossible to obtain. PMID- 11866588 TI - An effective method for the discrimination of motional anisotropy and chemical exchange. AB - Analysis of the ratio of transverse and longitudinal relaxation rates (R2/R1) is an approach commonly used for estimation of overall correlation time and identification of chemical exchange in biological macromolecules. However, this analysis fails to distinguish between chemical exchange and motional anisotropy. We describe a simple method for identifying chemical exchange and motional anisotropy using the product, R1R2. In the slow tumbling regime, the R1R2 product results in a constant value that is independent of overall correlation time and motional anisotropy. This analysis provides a simple method for rapidly estimating and dissociating the effects of motional anisotropy and chemical exchange in NMR heteronuclear spin relaxation data. We demonstrate the utility of the method with 15N relaxation data collected on the proteins E. coli ribonuclease H and the trimeric E. coli membrane associated lipoprotein lpp. PMID- 11866589 TI - Diazo group electrophilicity in kinamycins and lomaiviticin A: potential insights into the molecular mechanism of antibacterial and antitumor activity. AB - Theoretical and chemical studies of the reactivity of isoprekinamycin, the kinamycins, and the lomaiviticins support the proposal that these natural products exhibit enhanced diazonium salt character and may owe their antitumor antibiotic properties to their ability to act as electrophilic azo-coupling agents in vivo. PMID- 11866590 TI - The synthesis and biological characterization of a ceramide library. AB - A facile synthesis of a combinatorial ceramide library and their activities in the NF-kappaB pathway and in apoptosis induction/prevention were demonstrated. A novel NF-kappaB activating molecule was discovered among ceramide containing beta galactose, and the structural requirements of ceramides for apoptosis induction was elucidated. PMID- 11866591 TI - Can chiral single walled carbon nanotubes be used as enantiospecific adsorbents? AB - Single-walled carbon nanotubes can exist in chiral forms and can adsorb a range of molecules. We use atomistic simulations to consider whether enantiopure carbon nanotubes might be effective enantiospecific adsorbents for chiral molecules. We examine the adsorption of both enantiomers of trans-1,2-dimethylcyclopropane and trans-1,2-dimethylcyclohexane in a range of chiral nanotubes. Our simulations indicate that these molecules are strongly adsorbed in nanotubes, that is, they have large heats of adsorption, but the energy differences between adsorbed enantiomers are negligible. We argue that this result is generic for chiral organic molecules adsorbed in carbon nanotubes, suggesting that these materials will not be effective enantiospecific adsorbents. PMID- 11866592 TI - Unexpected substituent effects in offset pi-pi stacked interactions in water. AB - We have measured the rotational barriers of meta- and para-substituted N-benzyl-2 (2-fluorophenyl)pyridinium bromides in aqueous solution by dynamic NMR as a model system for offset-stacking interactions in proteins. Because the benzyl ring can stack with the 2-fluorophenyl ring in the offset conformation in the ground state, but not the transition state, the rotational barrier reflects the magnitude of the stacking interaction. Only a small (0.1 kcal/mol) change in rotational barrier was found for para substituents relative to hydrogen. A much larger energy difference was found for electronegative meta substituents (up to 0.66 kcal/mol for CF3). Evidence suggests that this is due at least in part to an electrostatic interaction between electron-poor hydrogens on one ring with the electronegative substituents on the other ring. PMID- 11866594 TI - Catalytic dealkylation of phosphates with binuclear boron compounds. AB - The Salen(tBu) ligand and its derivatives were used to prepare binuclear boron complexes. These compounds have the formula, L(BBr2)2 (L = Salpen(tBu) and Salben(tBu)). These are formed from the reaction of the corresponding L[B(OMe)2]2 with BBr3. They represent a new type of binuclear boron compound. These compounds are active towards the dealkylation of many phosphates. They are also catalytically active with a stoichiometric amount of BBr3 to trimethylphosphate. PMID- 11866593 TI - A rationally designed genotoxin that selectively destroys estrogen receptor positive breast cancer cells. AB - We describe a novel strategy to increase the selective toxicity of genotoxic compounds. The strategy involves the synthesis of bifunctional molecules capable of forming DNA adducts that have high affinity for specific proteins in target cells. It is proposed that the association of such proteins with damaged sites in DNA can compromise protein function and/or DNA repair resulting in increased toxicity. We describe the synthesis of a bifunctional compound consisting of an aniline mustard linked to the 7alpha position of estradiol. This novel compound can form covalent DNA adducts that have high affinity for the estrogen receptor. Breast cancer cells that express high levels of the estrogen receptor showed increased sensitivity to the cytotoxic effects of the new compound. PMID- 11866596 TI - Genius: a genetic algorithm for automated structure elucidation from 13C NMR spectra. AB - The automated structure elucidation of organic molecules from experimentally obtained properties is extended by an entirely new approach. A genetic algorithm is implemented that uses molecular constitution structures as individuals. With this approach, the structure of organic molecules can be optimized to meet experimental criteria, if in addition a fast and accurate method for the prediction of the used physical or chemical features is available. This is demonstrated using 13C NMR spectrum as readily obtainable information. By means of artificial neural networks a fast and accurate method for calculating the 13C NMR spectrum of the generated structures exists. The method is implemented and tested successfully for organic molecules with up to 18 non-hydrogen atoms. PMID- 11866597 TI - Nonaqueous biocatalytic synthesis of new cytotoxic doxorubicin derivatives: exploiting unexpected differences in the regioselectivity of salt-activated and solubilized subtilisin. AB - Two enzymes, Mucor javanicus lipase and subtilisin Carlsberg (SC), catalyzed the nonaqueous acylation of doxorubicin (DOX). Compared to the untreated enzyme the rate of DOX acylation at the C-14 position with vinyl butyrate in toluene was 25 fold higher by lipase ion-paired with Aerosol OT (AOT) and 5-fold higher by lipase activated by 98% (w/w) KCl co-lyophilization (3.21 and 0.67 mumol/min g lipase, respectively, vs 0.13 mumol/min g-lipase). Particulate subtilisin Carlsberg (SC) was nearly incapable of DOX acylation, but ion-paired SC (AOT-SC) catalyzed acylation at a rate of 2.85 mumol/min g-protease. The M. javanicus formulations, AOT-SC, and SC exclusively acylated the C14 primary hydroxyl group of DOX. Co-lyophilization of SC with 98% (w/w) KCl expanded the enzyme's regiospecificity such that KCl-SC additionally acylated the C4' hydroxyl and C3' amine groups. The total rate of DOX conversion with KCl-SC was 56.7 mumol/min g protease. The altered specificity of KCl-SC is a new property of the enzyme imparted by the salt activation, and represents the first report of unnatural regioselectivity exhibited by a salt-activated enzyme. Using AOT-SC catalysis, four unique selectively acylated DOX analogues were generated, and KCl-SC was used to prepare DOX derivatives acylated at the alternative sites. Cytotoxicities of select derivatives were evaluated against the MCF7 breast cancer cell line (DOX IC50 = 27 nM) and its multidrug-resistant sub-line, MCF7-ADR (DOX IC50 = 27 muM). The novel derivative 14-(2-thiophene acetate) DOX was relatively potent against both cell lines (IC50 of 65 nM and 8 muM, respectively) and the 14 (benzyl carbonate) DOX analogue was as potent as DOX against the MCF7 line (25 nM). Activated biocatalysts and their novel regioselectivity differences thus enabled single-step reaction pathways to an effective collection of doxorubicin analogues. PMID- 11866595 TI - A highly enantioselective route to either enantiomer of both alpha- and beta amino acid derivatives. AB - This report describes the unprecedented use of unmodified aldehydes as donors in a catalytic asymmetric Mannich-type reaction. The proline-catalyzed reaction of N PMP-protected alpha-imino ethyl glyoxylate with unmodified aliphatic aldehydes provided a general and very mild entry to either enantiomer of beta-amino and alpha-amino acids and derivatives in high yield and stereoselectivity. Six of the seven aldehydes studied yielded products with ee values of 99% or greater. The diastereoselectivity of the reaction increased with the bulkiness of the substituents of the aldehyde donor in the order R = Me < Et < i-Pr < n-Pent. In five of the cases studied, excellent syn stereoselectivities were achieved. In addition, the corresponding chiral beta-amino aldehyde adducts can be readily converted to the corresponding amino acid derivatives. Most significantly, this approach provides facile access to substituted beta-lactams. PMID- 11866598 TI - Photoregulation of DNA triplex formation by azobenzene. AB - Formation and dissociation of DNA triplex are reversibly photoregulated by cis <- > trans isomerization of the azobenzene tethered to the third strand. When the azobenzene takes the trans from, a stable triplex is formed. Upon the isomerization of trans-azobenzene to its cis form by UV light irradiation (300 < lambda < 400 nm), however, the modified oligonucleotide is removed from the target duplex. The triplex is re-formed on photoinduced cis --> trans isomerization (lambda > 400 nm). The photoregulating activity significantly depends on the position of azobenzene in the third strand, as well as on the geometric position (meta or para) of its amido substituent. For m amidoazobenzene, the photoregulation is the most effective when it is tethered to the 5'-end of the third strand. However, p-amidoazobenzene should be introduced into the middle of the strand for effective regulation. In the optimal cases, the change of T(m) of the triplex, caused by the cis <--> trans isomerization of azobenzene, is greater than 30 degrees C. UV-visible and CD spectroscopy, as well as computer modeling studies, clearly demonstrate that the trans-azobenzene intercalates between the base pairs in the target duplex and thus stabilizes the triplex by stacking interactions. On the other hand, nonplanar cis-azobenzene destabilizes the triplex due to its steric hindrance against the adjacent base pairs. PMID- 11866599 TI - Influence of backbone conformation on protein aggregation. AB - Effect(s) of organic solvents on an all beta-sheet protein are investigated to understand the influence of backbone conformation on protein aggregation. Results obtained in the present study reveal that protein aggregation is accompanied by the formation of non-native beta-sheet conformation. In contrast, induction of non-native helical segments in the protein is found to inhibit aggregation. The differential effects of the secondary structures on protein aggregation are proposed to stem from the disparity in the nature of the hydrogen bonds and packing of the side chains of hydrophobic residues in the beta-sheet and alpha helix conformation. In our opinion, the results of the present study provide useful hints to develop methods to alleviate the problems of both in vitro and in vivo protein aggregation. PMID- 11866601 TI - Efficient photoacids based upon triarylamine dialkylsulfonium salts. AB - New triarylamine dialkylsulfonium salts that are photosensitive in the near ultraviolet have been prepared. The quantum yields of photoacid generation were found to be approximately 0.5 and are independent of the counterion. On the other hand, the efficiencies of the sulfonium salts toward the photopolymerization of cyclohexene oxide depend on the counterion and the sulfonium substituents. Photopolymerization kinetic studies demonstrate that these triphenylamine sulfonium salts are highly efficient cationic photoinitiators. PMID- 11866600 TI - An experimental and computational investigation of the enantioselective deprotonation of Boc-piperidine. AB - The asymmetric deprotonation of N-Boc-piperidine (3) by the 1:1 complex of a sec alkyllithium and (-)-sparteine has been investigated both experimentally and computationally. The lithiation of 3 with sec-BuL-(-)-sparteine at -78 degrees C, which is a much slower process than is the analogous deprotonation of N-Boc pyrrolidine (1) and a minor reaction relative to the competing addition of sec BuLi to the carbamate, proceeds with a moderate degree of selectivity (er = 87:13) for removal of the pro-S hydrogen of 3. The related deprotonation of N-Boc 4-tosyloxypiperidine (6) with two molar equiv of sec-BuL-(-)-sparteine also involves preferential transfer of the pro-S hydrogen. The computational study of the deprotonation of (3) by i-PrL-(-)-sparteine found that the proton that is preferentially transferred within three-component intermediate complex is the thermodynamically least acidic alpha-hydrogen of 3. The asymmetric deprotonation of 3 is calculated to proceed with poor enantioselectivity and to have an activation energy considerably higher than that calculated for deprotonation of N Boc-pyrrolidine (1). The experimental and computational results are in good agreement. PMID- 11866602 TI - Quantitative evaluation of weak nonbonded Se...F interactions and their remarkable nature as orbital interactions. AB - To evaluate weak intramolecular nonbonded Se...F interactions recently characterized for a series of o-selenobenzyl fluoride derivatives (Iwaoka et al., Chem. Lett. 1998, 969-970), the temperature dependence of the nuclear spin coupling between Se and F (J(Se...F)) was investigated for 2 (fluoromethyl)phenylselenenyl cyanate (1a) and bis[2-(fluoromethyl)phenyl] diselenide (1e) in CD2Cl2 and CD3CN. A significant increase in the magnitude of J(Se...F) was observed for both 1a and 1e upon lowering temperature, whereas the values of J(Se...F) for the corresponding trifluoromethyl compounds slightly reduced or remained unchanged at low temperatures. Application of the rapid equilibrium model between two possible conformers revealed that conformer A with an intramolecular Se...F interaction is more stable in enthalpy (DeltaH) by 1.23 kcal/mol for 1a (in CD2Cl2) and by 0.85 and 0.83 kcal/mol for 1e (in CD2Cl2 and CD3CN, respectively) than conformer B, which does not have close Se...F contact. The negligible solvent effects for 1e suggested marginal electrostatic nature of the Se...F interactions. Instead, importance of the n(F) -->sigma*(Se-X) orbital interaction was suggested by quantum chemical (QC) calculations and the natural bond orbital (NBO) analysis. PMID- 11866603 TI - Bioinspired growth of crystalline carbonate apatite on biodegradable polymer substrata. AB - Mineralization in biological systems is a widespread, yet incompletely understood phenomenon involving complex interactions at the biomacromolecule-mineral nucleus interface. This study was aimed at understanding and controlling mineral formation in a poly(alpha-hydroxy ester) model system, to gain insight into biological mineralization processes and to develop biomaterials for orthopaedic tissue regeneration. We specifically hypothesized that providing a high surface density of anionic functional groups would enhance nucleation and growth of bonelike mineral following exposure to simulated body fluids (SBF). Polymer surface functionalization was achieved via hydrolysis of 85:15 poly(lactide-co glycolide) (PLG) films. This treatment led to an increase in surface carboxylic acid and hydroxyl groups, resulting in a substantial increase in polymer surface energy from 42 to 49 dynes/cm2. Treated polymers exhibited a 3-fold increase in heterogeneous mineral grown and growth of a continuous mineral film on the polymer surface. The mineral grown on PLG surfaces is a carbonate apatite, the major mineral component of vertebrate bone tissue. Mineral crystal size and morphology were dependent on the solution characteristics but unaffected by the degree of surface prehydrolysis. The mechanism of heterogeneous carbonate apatite growth was examined via ion binding assays, which indicated that calcium binding is mediated independently by the presence of soluble phosphate counterions and surface functional groups. These findings indicate that poly(alpha-hydroxy ester) materials can be readily mineralized using a biomimetic process, and that the impetus for mineral nucleation in this system appears more complicated than the simple electrostatic interactions proposed in previous biomineralization theory. PMID- 11866604 TI - Measurement of the hydride donor abilities of [HM(diphosphine)2]+ complexes (M = Ni, Pt) by heterolytic activation of hydrogen. AB - [M(diphosphine)2]2+ complexes (where M = Ni and Pt) react with hydrogen in the presence of bases to form the corresponding hydrides, [HM(diphosphine)2]+. In seven cases, equilibria have been observed from which the hydride donor ability (DeltaGdegrees(H-)) of the hydrides can be calculated. For six of these complexes, the DeltaGdegrees(H-) values calculated using heterolytic activation of hydrogen are compared with those based on thermodynamic cycles using pK(a) measurements and electrochemical half-wave potentials. The agreement between these two methods is good (within 1 kcal/mol). The reactivity of the various [M(diphosphine)2]2+ complexes toward hydrogen parallels their measured hydride acceptor abilities. PMID- 11866605 TI - Hydride donor abilities and bond dissociation free energies of transition metal formyl complexes. AB - The hydride complex [Pt(dmpe)2H]+ (dmpe = 1,2-bis(dimethylphosphino)ethane) reversibly transfers H- to the rhenium carbonyl complex [CpRe(PMe3)(NO)(CO)]+, giving the formyl CpRe(PMe3)(NO)(CHO). From the equilibrium constant for the hydride transfer (16.2), the DeltaGdegrees for the reaction was determined (-1.6 kcal/mol), as was the hydride-donating ability of the formyl (44.1 kcal/mol). The hydride-donating ability, DeltaGdegrees(H-), is defined as the energy required to release the hydride ion into solution by the formyl complex [i.e. M(CHO) right arrow M(CO)+ + H-]. Subsequently, the hydride-donating ability of a series of formyl complexes was determined, ranging from 44 to 55 kcal/mol. With use of this information, two rhenium carbonyl complexes, [CpRe(NO)(CO)2]+ and [Cp*Re(NO)(CO)2]+, were hydrogenated to formyls, employing [Pt(dmpp)2]2+ and Proton-Sponge. Finally, the E(1/2)(I/0) values for five rhenium carbonyl complexes were measured by cyclic voltammetry. Combined with the known DeltaGdegrees(H-) values for the complexes, the hydrogen atom donating abilities could be determined. These values were all found to be approximately 50 kcal/mol. PMID- 11866606 TI - Interaction of biotin with Mg-O bonds: bifunctional binding and recognition of biotin and related ligands by the Mg(15-crown-5)2+ unit. AB - The interaction between biotin and the macrocyclic magnesium complex Mg(15-crown 5)(Otf)2 (15-crown-5 is 1,4,7,10,13-pentaoxacyclopentadecane, Otf(-) is trifluoromethanesulfonate anion) in solution was studied as a model for metal biotin interactions that may be important in its speciation and function. Shifts in the solution IR spectrum establish that the interaction is dominated by ligation between the carbonyl oxygen of the ureido ring of biotin and the Mg2+ cation. However, comparative binding studies using NMR spectroscopy and conductivity reveal a substantial enthalpic contribution to binding that arises from interactions between the ureido -NH moiety and the macrocyclic ring. This is interpreted in terms of a weak-to-moderate hydrogen bond formed between the -NH group and an oxygen from the crown, which is simultaneously coordinated to Mg2+. This hypothesis is reinforced by quantitative examination of the binding of N methylated derivatives of 2-imidazolidone, which shows that N,N'-dimethylation decreases the affinity of Mg(15-crown-5)(Otf)2 for the ligand by 2 orders of magnitude. This can be understood in terms of the structure of Mg(15-crown 5)(Otf)2. It shows a pentagonal bipyramidal coordination geometry where the five equatorial positions are occupied by the macrocyclic oxygen donors. The axial positions are occupied by weakly coordinating Otf(-) anions, which are readily displaced by biotin and related derivatives. The M-O(crown) bond distance ranges from 2.1 to 2.3 A, providing structural complementarity for the 2.2 A C=O...HN- bite distance in the ureido group, which leads to strong interaction. The contribution from hydrogen bonding illustrates the importance of second-shell interactions in the biocoordination chemistry of Mg2+. These can serve to organize cofactor interactions with biomolecules, as was recently demonstrated for a biotin-selective RNA aptamer that depends on a direct biotin-magnesium interaction for recognition of biotin (Nix, J.; Sussman, D.; Wilson, C. J. Mol. Biol. 2000, 296, 1235-1244). These results are significant in the context of the observed magnesium requirement in biotin-dependent carboxylase enzymes, where noncovalent interactions with biotin may be important in its activation toward carboxylation in the first step of biotin-dependent CO2 transfer. The synthetic system presented here also suggests that the Mg-O bond may be considered a constituent design element in the rational preparation of complexes to bind and recognize biotin. PMID- 11866607 TI - Formation and characterization of carbon monoxide adducts of iron "twin coronet" porphyrins. Extremely low CO affinity and a strong negative polar effect on bound CO. AB - The carbon monoxide (CO) adducts of iron "twin coronet" porphyrins (TCPs) are characterized by UV-vis, resonance Raman (RR), IR, and 13C NMR spectroscopies. A superstructured porphyrin, designated as TCP, was used as a common framework for the four different types of iron complexes. TCP bears two binaphthalene bridges on each side and creates two hydrophobic pockets surrounded by the bulky aromatic rings. In the CO-binding cavities, the hydroxyl groups are oriented toward the center above the heme. The iron complexes investigated are as follows: TCP (which is without a covalently linked axial ligand), TCP-PY (which has a linked pyridine ligand), and TCP-TB and TCP-TG (both of which have a linked thiolate ligand). These complexes were synthesized as ferric forms and identified by the various spectroscopic methods. The UV-vis spectra of TCP-CO and TCP-PY-CO exhibit lambda(max) at 432, 546 and 428, 541 nm, respectively. On the other hand, the CO adducts of TCP-TB and TCP-TG show typical hyperporphyrin spectra for a thiolate ligated iron(II) porphyrin-CO complex. In the RR spectra, the nu(Fe-CO) bands were observed at 506, 489 cm(-1) (TCP), 465 cm(-1) (TCP-PY), 458, 437 cm(-1) (TCP TG) and 429 cm(-1) (TCP-TB). Compared with the reported nu(Fe-CO) frequencies of hemoproteins and their model systems, these observed values are unusually low. Further, abnormally high nu(C-O) bands are observed at 1990 cm(-1) (TCP-CO) and 2008 cm(-1) (TCP-PY-CO) in IR spectra. The lower nu(Fe-CO) and the higher nu(C-O) frequencies can be ascribed to the strong negative polar effect caused by the vicinal hydroxyl groups in the cavity. This prediction is further supported by the observation of significant 13C shieldings exhibited by TCP-CO (delta = 202.6 ppm) and TCP-PY-CO (delta = 202.3 ppm), in comparison to hemoproteins and other heme models. The CO affinity of TCP-PY (P1/2CO = 0.017 Torr at 25 C) is unusually lower than other heme models. The unique behavior of these CO adducts is discussed in context of the TCP structures. PMID- 11866608 TI - Structural modulation of molybdenyl iodate architectures by alkali metal cations in AMoO3(IO3) (A = K, Rb, Cs): a facile route to new polar materials with large SHG responses. AB - Three new molybdenyl iodates, KMoO3(IO3) (1), RbMoO3(IO3) (2), and CsMoO3(IO3) (3), have been prepared through the hydrothermal reactions of MoO3 with AIO4 (A = K, Rb, or Cs) at 180 C. These compounds are isolated as nearly colorless, air stable crystals. Single-crystal X-ray diffraction experiments reveal that 1 possesses a corrugated layered structure constructed from molybdenum oxide chains that are bridged by iodate anions. The puckering of the layers is caused by the alignment of bent molybdenyl (MoO2(2+)) groups along one side of the molybdenum oxide chains. The K+ cations separate these layers from one another and serve to balance charge. In contrast, compounds 2 and 3, which are isostructural, form three-dimensional structures with small cavities filled with Rb+ or Cs+ cations. The differences between the structures of 1 and those of 2 and 3 are due to rotation of the molybdenyl units as translation occurs down the molybdenum oxide chains in order to accommodate the increased size of the Rb+ and Cs+ cations. This rotation allows for the iodate anions to bridge the molybdenum oxide chains in an additional dimension, creating a three-dimensional network structure. Furthermore, while 1 crystallizes in a centrosymmetric space group, 2 and 3 crystallize in polar space groups. Second-harmonic generation measurements on 2 and 3 show large responses of 400x alpha-quartz. Differential scanning calorimetry measurements demonstrate that 2 and 3 are thermally stable to 494 and 486 C, respectively. UV-vis diffuse reflectance spectra of these compounds show a high degree of transparency from 1 to 3 eV and a band gap of 3.1 eV. PMID- 11866609 TI - Hydrogen and higher shell contributions in Zn2+, Ni2+, and Co2+ aqueous solutions: an X-ray absorption fine structure and molecular dynamics study. AB - A detailed investigation of the hydration structure of Zn2+, Ni2+, and Co2+ in water solutions has been carried out combining X-ray absorption fine structure (EXAFS) spectroscopy and Molecular Dynamics (MD) simulations. The first quantitative analysis of EXAFS from hydrogen atoms in 3d transition metal ions in aqueous solutions has been carried out and the ion-hydrogen interactions have been found to provide a detectable contribution to the EXAFS spectra. An accurate determination of the structural parameters associated with the first hydration shell has been performed and compared with previous experimental results. No evidence of significant contributions from the second hydration shell to the EXAFS signal has been found for these solutions, while the inclusion of the hydrogen signal has been found to be important in performing a quantitative analysis of the experimental data. The high-frequency contribution present in the EXAFS spectra has been found to be due to multiple scattering (MS) effects inside the ion-oxygen first coordination shell. MD has been used to generate three-body distribution functions from which a reliable analysis of the MS contributions to the EXAFS spectra of these systems has been carried out. PMID- 11866610 TI - Development and validation of an integrated computational approach for the study of ionic species in solution by means of effective two-body potentials. The case of Zn2+, Ni2+, and Co2+ in aqueous solutions. AB - In this paper we have developed an effective computational procedure for the structural and dynamical investigation of ions in aqueous solutions. Quantum mechanical potential energy surfaces for the interaction of a transition metal ion with a water molecule have been calculated taking into account the effect of bulk solvent by the polarizable continuum model (PCM). The effective ion-water interactions have been fitted by suitable analytical potentials, and have been utilized in molecular dynamics (MD) simulations to obtain structural and dynamical properties of the ionic aqueous solutions. This procedure has been successfully applied to the Co2+-H2O open-shell system and, for the first time, Co-oxygen and Co-hydrogen pair potential functions have been determined and employed in MD simulations. The reliability of the whole procedure has been assessed by applying it also to the Zn2+ and Ni2+ aqueous solutions, and the structural and dynamical properties of the three systems have been calculated by means of MD simulations and have been found to be in very good agreement with experimental results. The structural parameters of the first solvation shells issuing from the MD simulations provide an effective complement to extended X-ray absorption fine structure (EXAFS) experiments. PMID- 11866611 TI - Preparation and photophysical studies of a fluorous phase-soluble fullerene derivative. AB - We report the first synthesis of a well-characterized "Teflon ponytail" fullerene adduct (3) via the Hirsch-Bingel reaction with a malonate bearing two perfluorinated alkyl chains. This C3 tris-adduct shows excellent solubility in perfluorinated solvents, such as FC-72 and FC-75. Compound 3 was found to be an efficient sensitizer for singlet oxygen formation in fluorous media, which has potential in biphasic systems and in photobiology. PMID- 11866612 TI - Modern spectrum analysis in multidimensional NMR spectroscopy: comparison of linear-prediction extrapolation and maximum-entropy reconstruction. AB - NMR spectroscopy is an inherently insensitive technique, and many challenging applications such as biomolecular studies operate at the very limits of sensitivity and resolution. Advances in superconducting magnet, cryogenic probe, and pulse sequence technologies have resulted in dramatic improvements in both sensitivity and resolution in the past decade. Conversely, the signal-processing method used most widely in NMR spectroscopy, extrapolation of the time domain signal by linear prediction (LP) followed by discrete Fourier transformation (DFT), was developed in the early 1980s and has not been subjected to detailed scrutiny for its impact on sensitivity and resolution. Here we report the first systematic investigation of the accuracy and precision of spectra obtained by LP extrapolation followed by DFT. We compare the results to spectra obtained by maximum-entropy (MaxEnt) reconstruction, which was developed contemporaneously to LP extrapolation but is not widely employed in NMR spectroscopy. Although it reduces truncation artifacts and increases the amplitudes of strong peaks, we find that LP extrapolation generates false-positive peaks and introduces frequency errors. These defects of LP extrapolation become less pronounced for longer data records and higher signal-to-noise ratio. MaxEnt generally yields more detectable peaks for a given number of data samples, more accurate peak frequencies, and fewer false-positive peaks than LP extrapolation. MaxEnt also permits the use of nonlinear sampling, which can give dramatic improvements in resolution. These results show that the use of MaxEnt together with nonlinear sampling, rather than LP extrapolation, could reduce the amount of instrument time required for adequate sensitivity and resolution by a factor of 2 or more. PMID- 11866613 TI - Production, analysis, and application of spatially resolved shells in solid-phase polymer spheres. AB - Two-photon fluorescence microscopy has been used to interrogate the interior functionality of polymer resin beads. By employing this technique, the spatial distribution of the initial functionality contained within the polymer matrix has been determined. Spatially resolved, concentric shells were then produced synthetically in these polymer spheres via a series of protection/deprotection reactions in which two-photon fluorescence microscopy was employed to monitor each successive step. To demonstrate the potential utility of these techniques in combinatorial screening, a set of beads was prepared containing a unique tripeptide sequence in each of the three concentric shells within each individual bead. The set was then screened for the binding affinity of each tripeptide toward a fluorescent ligand. PMID- 11866614 TI - Multichannel digital transmission in an optical network of communicating molecules. AB - In present telecommunication networks, information transfer relies on the interplay of optical and electrical signals. Data are communicated optically but processed electronically. Methods to maintain the propagating signals solely at the optical level must be developed to overcome the transmission capacities and speed limits imposed by the electronic components. We have demonstrated that molecular switches can be used to gate optical signals in response to optical signals. We have realized a simple optical network consisting of three light sources, one cell containing a solution of three fluorescent molecules, one cell containing a solution of a three-state molecular switch and a detector. The light emitted by the three fluorophores is absorbed by the three states of the molecular switch. Using this simple operating principle, we have shown that multichannel digital transmission can be implemented on an ensemble of communicating molecules relying exclusively on the interplay of optical inputs and optical outputs. PMID- 11866615 TI - The leap-frog effect of ring currents in benzene. AB - Symmetry arguments show that the ring-current model proposed by Pauling, Lonsdale, and London to explain the enhanced diamagnetism of benzene is flawed by an intrinsic drawback. The minimal basis set of six atomic 2p orbitals taken into account to develop such a model is inherently insufficient to predict a paramagnetic contribution to the perpendicular component of magnetic susceptibility in planar ring systems such as benzene. Analogous considerations can be made for the hypothetical H(6) cyclic molecule. A model allowing for extended basis sets is necessary to rationalize the magnetism of aromatics. According to high-quality coupled Hartree-Fock calculations, the trajectories of the current density vector field induced by a magnetic field perpendicular to the skeletal plane of benzene in the pi electrons are noticeably different from those typical of a Larmor diamagnetic circulation, in that (i) significant deformation of the orbits from circular to hexagonal symmetry occurs, which is responsible for a paramagnetic contribution of pi electrons to the out-of-plane component of susceptibility, and (ii) a sizable component of the pi current density vector parallel to the inducing field is predicted. This causes a waving motion of pi electrons; streamlines are characterized by a "leap-frog effect". PMID- 11866616 TI - Efficient photocyclization of dithienylethene dimer, trimer, and tetramer: quantum yield and reaction dynamics. AB - Multi-dithienylethene arrays, in which two, three, or four 1,2-bis(2,4 dimethylthiophen-3-yl)perfluorocyclopentenes are ethynylene-bridged, were synthesized. Upon irradiation with ultraviolet light the hexane solutions of the arrays turned violet-blue and the color disappeared by irradiation with visible light. The quantum yields of photocyclization reactions successively increased from 0.21 to 0.40 by increasing the number of the dithienylethene moieties in the arrays from one to four. Picosecond laser photolysis as well as the fluorescence depolarization experiment confirmed that efficient excited energy migration in the arrays from the photochemically inactive parallel conformer to the photoactive antiparallel conformer resulted in the high quantum yields. PMID- 11866617 TI - pH and pK determinations by high-resolution solid-state 13C NMR: acid-base and tautomeric equilibria of lyophilized L-histidine. AB - Acid-base properties of lyophilized powders of L-histidine have been systematically investigated using parent solutions at pH varying from 1.8 to 10. For the first time, high-resolution solid-state 13C NMR was shown to allow separate observation of all three acid-base pairs in the successive deprotonations of the carboxylic end, the imidazolium cation, and the terminal ammonio group of histidine. 1H CRAMPS NMR spectra directly visualize the absence of the N3-H(pi) tautomer in neutral and anionic species. Solid-state titration shifts are enlarged by approximately 1-4 ppm with respect to those measured in solution, permitting unambiguous observation of conjugate acid-base pairs. Calculated pK's from solid-state acid-to-base ratios r are found equal to those classically measured in solution at 0 degrees C with a similar ionic strength of 0.1 mol x dm(-3). This proves that natural-abundance 13C solid-state determinations of r can be used to measure pK's in parent solutions without recourse to full titration curves and subsequent curve-fitting procedures. Such an approach also leads to noninvasive characterizations of the acidity of lyophilized powders, i.e., to the prediction of in situ pH of products obtained after rehydration and solubilization of powders. These results show the possibility of measuring the pK of nonvolatile acidic substrates dissolved in any sublimable solvent through lyophilization of the investigated solutions; this leads the way to pH and pK determinations when electrochemical or spectrophotometric measurements are impossible or ambiguous, e.g., for concentrated solutions, polyacids, or mixtures of acidic solutes, and possibly to the establishment of pK scales in nonaqueous solvents and in melts. PMID- 11866618 TI - An ab initio quantum-chemical study of the blue-copper site of azurin. AB - The electronic structure of the blue-copper site of Pseudomonas aeruginosa azurin has been investigated by ab initio multireference determinantal configuration interaction (MRD-CI) calculations. A truncated site consisting of copper and its three equatorial ligands has been studied with emphasis on the g tensor and the nitrogen hyperfine tensors of the coordinating histidines. In the ground state the singly occupied molecular orbital (SOMO) involves a copper 3d orbital pi antibonded to the cysteine sulfur and sigma antibonded to the histidine nitrogens. A proper description of the electron-paramagnetic-resonance parameters has been achieved through the use of an effective core potential for copper up to and including the 3s electrons. Both the complete g tensor and the anisotropic hyperfine tensors at the nitrogens are essentially reproduced. Mulliken spin densities of 35 and 59% on copper and sulfur, respectively, and 2.1 and 1.7% on the respective coordinating nitrogens reflect the delocalized character of the SOMO and the inequivalence of the histidines. PMID- 11866619 TI - Multiexponential electronic spin relaxation and Redfield's limit in Gd(III) complexes in solution: consequences for 17O/1H NMR and EPR simultaneous analysis. AB - Multiple experiments (17O NMR, 1H NMR, and EPR) have been performed in the past to understand the microscopic parameters that control the magnetic relaxation rate enhancement induced by paramagnetic molecules on neighboring water protons, the so-called relaxivity. The generally accepted theories of the electron spin relaxation of S = 7/2 ions such as Gd3+ (Solomon-Bloembergen-Morgan or simplified Hudson-Lewis) are unsatisfactory for a simultaneous analysis. Recently, an improved theory, where the electron spin relaxation is due to the combination of a static (thus explicitly linked to the molecular structure) and a dynamic zero field splitting, has been developed and tested on experimental EPR data. The model has also been extended beyond the electronic Redfield limit using Monte Carlo simulations. Using the aqua ion [Gd(H2O)8]3+ as a test case, we present here the first simultaneous analysis of 17O NMR, 1H NMR, and EPR relaxation data using this rigorous approach of the electron spin relaxation. We discuss the physical meaning of the calculated parameters. The consequences on future experiments are also considered, especially regarding the analysis of nuclear magnetic relaxation dispersion (NMRD) profiles in the study of Gd3+ complexes. PMID- 11866620 TI - Control of photoluminescence properties of CdSe nanocrystals in growth. AB - The photoluminescence (PL) quantum yield (QY) of CdSe nanocrystals during their growth under a given set of initial conditions increases monotonically to a certain maximum value and then decreases gradually. Such a maximum is denoted as a PL "bright point", which does not always overlap with the minimum point of the PL peak width for the same reaction. The experimental results suggest that the existence of the PL bright point is a general phenomenon during the growth of semiconductor nanocrystals and likely is a signature of an optimal surface structure/reconstruction of the nanocrystals grown under a given set of initial conditions. The position of the bright point, the highest PL QY, the types of the bright points (sharp or flat), the sharpness of the PL peak, etc., were all strongly dependent on the initial Cd:Se ratio of the precursors in the solution. A large excess of the selenium precursor, with 5-10 times more selenium precursor than the amount of the cadmium precursor, was found necessary to achieve a high PL QY value and a narrow emission profile. The existence of the PL bright point and the sensitive temporal variation of the PL QY during the growth of semiconductor nanocrystals can explain the unpredictable nature and poor reproducibility of the PL properties of the as-prepared semiconductor nanocrystals observed previously. Furthermore, the knowledge gained in this study enabled us to reproducibly synthesize highly luminescent CdSe nanocrystals through a relatively simple and safe synthetic scheme. In a traditionally weak emission window for CdSe nanocrystals, the orange-red optical window, the PL QY of the as-prepared CdSe nanocrystals reached as high as 85% at room temperature, and the full width at half-maximum of the corresponding PL peak was as narrow as 23 nm, about 65-80 meV depending on the emitting position. The PL properties of the as-prepared CdSe nanocrystals are stable upon aging for at least several months. These as-prepared nanocrystals represent a series of best emitters that are highly efficient, highly pure in emission color, stable, and continuously tunable by simply varying the size of the nanocrystals. PMID- 11866621 TI - Noncovalent binding of sensitizers for lanthanide(III) luminescence in an EDTA bis(beta-cyclodextrin) ligand. AB - EDTA-linked beta-cyclodextrin dimer 3 was synthesized from EDTA bis(anhydride) 1 and mono(propylamino)-appended beta-cyclodextrin 2. p-tert-butylbenzoate 5, bound by the beta-cyclodextrin cavities of 3 with an association constant of 10(4) M( 1) in water, acts as a sensitizer for the Eu(III) and Tb(III) complexes of 3. Luminescence spectroscopy, microcalorimetry, and Gd(III)-induced NMR relaxation rate measurements prove that 3 forms a 1:2 complex with 5 and that one of the beta-cyclodextrin-bound sensitizers coordinates to the EDTA-encapsulated Ln(III) ion. The Eu(III) complex of 3 forms strong 1:1 complexes (K approximately 10(7) M(-1)) with bis(propylamido adamantyl)-functionalized biphenyl sensitizers 7 and 8 in water. Both beta-cyclodextrins of 3 are involved in the binding of these guests. The amide functionality adjacent to the biphenyl unit in 7 and 8 coordinates to the EDTA-encapsulated Ln(III) ion. For these biphenyl-based antennae both binding to beta-cyclodextrin and coordination to the Ln(III) center are crucial for efficient sensitization. PMID- 11866622 TI - The aromaticity of pyracylene: an experimental and computational study of the energetics of the hydrogenation of acenaphthylene and pyracylene. AB - In this work, the aromaticity of pyracylene (2) was investigated from an energetic point of view. The standard enthalpy of hydrogenation of acenaphthylene (1) to acenaphthene (3) at 298.15 K was determined to be minus sign(114.5 +/- 4.2) kJ x mol(-1) in toluene solution and minus sign(107.9 +/- 4.2) kJ x mol(-1) in the gas phase, by combining results of combustion and reaction-solution calorimetry. A direct calorimetric measurement of the standard enthalpy of hydrogenation of pyracylene (2) to pyracene (4) in toluene at 298.15 K gave (249.9 plus minus 4.6) kJ x mol(-1). The corresponding enthalpy of hydrogenation in the gas phase, computed from the Delta(f)H(o)m(cr) and DeltaH(o)m(sub) values obtained in this work for 2 and 4, was -(236.0 +/- 7.0) kJ x mol(-1). Molecular mechanics calculations (MM3) led to Delta(hyd)H(o)m(1,g) = -110.9 kJ x mol(-1) and Delta(hyd)H(o)m(2,g) = -249.3 kJ x mol(-1) at 298.15 K. Density functional theory calculations [B3LYP/6-311+G(3d,2p)//B3LYP/6-31G(d)] provided Delta(hyd)H(o)m(2,g) = -(244.6 +/- 8.9) kJ x mol(-1) at 298.15 K. The results are put in perspective with discussions concerning the "aromaticity" of pyracylene. It is concluded that, on energetic grounds, pyracylene is a borderline case in terms of aromaticity/antiaromaticity character. PMID- 11866625 TI - Sleep deprivation and clinical performance. PMID- 11866627 TI - Guidelines offer "roadmap" for chronic kidney disease. PMID- 11866626 TI - Study concludes that moderate PSA levels are unrelated to prostate cancer outcomes. PMID- 11866632 TI - Risk factors for violent death in children. PMID- 11866635 TI - Evaluation of inhalational anthrax. PMID- 11866633 TI - Risk factors for violent death in children. PMID- 11866636 TI - Evaluation of inhalational anthrax. PMID- 11866638 TI - Usefulness of positron emission tomography in evaluating dementia. PMID- 11866639 TI - Usefulness of positron emission tomography in evaluating dementia. PMID- 11866641 TI - Patient safety in trials of therapy for acute ischemic stroke. PMID- 11866643 TI - Therapy for cocaine addiction. PMID- 11866644 TI - Poverty and responsibility for health care. PMID- 11866646 TI - Evaluation of investigations conducted to detect and prevent transmission of tuberculosis. AB - CONTEXT: Contact investigations are routinely conducted by health departments throughout the United States for all cases of active pulmonary tuberculosis (TB) to identify secondary cases of active TB and latent TB infection and to initiate therapy as needed in these contacts. Little is known about the actual procedures followed, or the results. OBJECTIVES: To evaluate contact investigations conducted by US health departments and the outcomes of these investigations. DESIGN, SETTING, AND SUBJECTS: Review of health department records for all contacts of 349 patients with culture-positive pulmonary TB aged 15 years or older reported from 5 study areas in the United States during 1996. MAIN OUTCOME MEASURES: Number of contacts identified, fully screened, and infected per TB patient; rates of TB infection and disease among contacts of TB patients; and type and completeness of data collected during contact investigations. RESULTS: A total of 3824 contacts were identified for 349 patients with active pulmonary TB. Of the TB patients, 45 (13%) had no contacts identified. Of the contacts, 55% completed screening, 27% had an initial but no postexposure tuberculin skin test, 12% were not screened, and 6% had a history of prior TB or prior positive tuberculin skin test. Of 2095 contacts who completed screening, 68% had negative skin test results, 24% had initial positive results with no prior test result available, 7% had documented skin test conversions, and 1% had active TB at the time of investigation. Close contacts younger than 15 years (76% screened vs 65% for older age groups; P<.001) or exposed to a TB patient with a positive smear (74% screened vs 59% for those with a negative smear; P<.001) were more likely to be fully screened. Close contacts exposed to TB patients with both a positive smear and a cavitary chest radiograph were more likely to have TB infection or disease (62% vs 33% for positive smear only vs 44% for cavitary radiograph only vs 37% for neither characteristic; P<.001). A number of factors associated with TB patient infectiousness, contact susceptibility to infection, contact risk of progression to active TB, and amount of contact exposure to the TB patient were not routinely recorded in health department records. CONCLUSIONS: Improvement is needed in the complex, multistep process of contact investigations to ensure that contacts of patients with active pulmonary TB are identified and appropriately screened. PMID- 11866647 TI - Predictive model to identify positive tuberculosis skin test results during contact investigations. AB - CONTEXT: Budgetary constraints in tuberculosis (TB) control programs require streamlining contact investigations without sacrificing disease control. OBJECTIVE: To develop more efficient methods of TB contact investigation by creating a model of TB transmission using variables that best predict a positive tuberculin skin test among contacts of an active TB case. DESIGN, SETTING, AND SUBJECTS: After standardizing the interview and documentation process, data were collected on 292 consecutive TB cases and their 2941 contacts identified by the Alabama Department of Public Health between January and October 1998. Generalized estimating equations were used to create a model for predicting positive skin test results in contacts of active TB cases. The model was then validated using data from a prospective cohort of 366 new TB cases and their 3162 contacts identified between October 1998 and April 2000. MAIN OUTCOME MEASURE: Tuberculin skin test result. RESULTS: Using generalized estimating equations to build a predictive model, 7 variables were found to significantly predict a positive tuberculin skin test result among contacts of an active TB case. Further testing showed this model to have a sensitivity, specificity, and positive predictive value of approximately 89%, 36%, and 26%, respectively. The false-negative rate was less than 10%, and about 40% of the contact workload could be eliminated using this model. CONCLUSIONS: Certain characteristics can be used to predict contacts most likely to have a positive tuberculin skin test result. Use of such models can significantly reduce the number of contacts that public health officials need to investigate while still maintaining excellent disease control. PMID- 11866648 TI - Residual lifetime risk for developing hypertension in middle-aged women and men: The Framingham Heart Study. AB - CONTEXT: The long-term risk for developing hypertension is best described by the lifetime risk statistic. The lifetime risk for hypertension and trends in this risk over time are unknown. OBJECTIVES: To estimate the residual lifetime risk for hypertension in older US adults and to evaluate temporal trends in this risk. DESIGN, SETTING, AND PARTICIPANTS: Community-based prospective cohort study of 1298 participants from the Framingham Heart Study who were aged 55 to 65 years and free of hypertension at baseline (1976-1998). MAIN OUTCOME MEASURES: Residual lifetime risk (lifetime cumulative incidence not adjusted for competing causes of mortality) for hypertension, defined as blood pressure of 140/90 mm Hg or greater or use of antihypertensive medications. RESULTS: The residual lifetime risks for developing hypertension and stage 1 high blood pressure or higher (greater-than or-equal to 140/90 mm Hg regardless of treatment) were 90% in both 55- and 65 year-old participants. The lifetime probability of receiving antihypertensive medication was 60%. The risk for hypertension remained unchanged for women, but it was approximately 60% higher for men in the contemporary 1976-1998 period compared with an earlier 1952-1975 period. In contrast, the residual lifetime risk for stage 2 high blood pressure or higher (greater-than-or-equal to 160/100 mm Hg regardless of treatment) was considerably lower in both sexes in the recent period (35%-57% in 1952-1975 vs 35%-44% in 1976-1998), likely due to a marked increase in treatment of individuals with substantially elevated blood pressure. CONCLUSION: The residual lifetime risk for hypertension for middle-aged and elderly individuals is 90%, indicating a huge public health burden. Although the decline in lifetime risk for stage 2 high blood pressure or higher represents a major achievement, efforts should be directed at the primary prevention of hypertension. PMID- 11866649 TI - Effectiveness of safety measures recommended for prevention of workplace homicide. AB - CONTEXT: Homicide is the second leading cause of death on the job for US workers. Government agencies recommend that employers prevent violence against workers by adopting interventions originally designed to prevent robbery, but the effectiveness of these interventions is unknown. OBJECTIVE: To investigate the effectiveness of existing administrative and environmental interventions recommended for preventing workplace homicide. DESIGN, SETTING, AND PARTICIPANTS: Population-based case-control study of North Carolina workplaces where a worker had been killed between January 1, 1994, and March 31, 1998, identified through a statewide medical examiner system (cases; n = 105) and an industry-matched random sample of workplaces at risk during the same period, selected from business telephone listings (controls; n = 210). MAIN OUTCOME MEASURE: Risk of death of a worker due to homicide. RESULTS: Among environmental interventions, strong and consistent reductions in the risk of a worker being killed on the job were associated with bright exterior lighting (odds ratio [OR], 0.5; 95% confidence interval [CI], 0.3-1.0). Among administrative interventions, the largest beneficial effect was for staffing practices that prevented workers from being alone at night (OR, 0.4; 95% CI, 0.2-0.9). Keeping doors closed during working hours was also associated consistently with substantially reduced risk (OR, 0.4; 95% CI, 0.1-1.1) but was not statistically significant. Combinations of 5 or more administrative measures were associated with significantly lower levels of risk (OR, 0.1; 95% CI, 0.0-0.5). CONCLUSIONS: We found evidence suggesting that eliminating solo work at night could reduce the risk of homicide for workers. Keeping doors closed and using bright exterior lighting or combinations of administrative interventions also appear to be beneficial, but there was no evidence of effectiveness for a number of other recommended measures. PMID- 11866650 TI - Preimplantation diagnosis for early-onset Alzheimer disease caused by V717L mutation. AB - CONTEXT: Indications for preimplantation genetic diagnosis (PGD) have recently been expanded to include disorders with genetic predisposition to allow only embryos free of predisposing genes to be preselected for transfer back to patients, with no potential for pregnancy termination. OBJECTIVE: To perform PGD for early-onset Alzheimer disease (AD), determined by nearly completely penetrant autosomal dominant mutation in the amyloid precursor protein (APP) gene. DESIGN: Analysis undertaken in 1999-2000 of DNA for the V717L mutation (valine to leucine substitution at codon 717) in the APP gene in the first and second polar bodies, obtained by sequential sampling of oocytes following in vitro fertilization, to preselect and transfer back to the patient only the embryos that resulted from mutation-free oocytes. SETTING: An in vitro fertilization center in Chicago, Ill. PATIENTS: A 30-year-old AD-asymptomatic woman with a V717L mutation that was identified by predictive testing of a family with a history of early-onset AD. MAIN OUTCOME MEASURES: Results of mutation analysis; pregnancy outcome. RESULTS: Four of 15 embryos tested for maternal mutation in 2 PGD cycles, originating from V717L mutation--free oocytes, were preselected for embryo transfer, yielding a clinical pregnancy and birth of a healthy child free of predisposing gene mutation according to chorionic villus sampling and testing of the neonate's blood. CONCLUSION: This is the first known PGD procedure for inherited early onset AD resulting in a clinical pregnancy and birth of a child free of inherited predisposition to early-onset AD. PMID- 11866651 TI - Home visits to prevent nursing home admission and functional decline in elderly people: systematic review and meta-regression analysis. AB - CONTEXT: The effects of home visitation programs to prevent functional decline in elderly persons have been inconsistent, and the value of these programs is controversial. OBJECTIVE: To evaluate the effect of preventive home visits on functional status, nursing home admission, and mortality. DATA SOURCES: Studies published in English, French, German, Italian, or Spanish reporting randomized trials of the effects of preventive in-home visits in older people (mean age >70 years) living in the community were identified through searches of MEDLINE, PSYCHINFO, and EMBASE (January 1985--November 2001). We also searched the Cochrane Controlled Trials Register, checked reference lists of earlier reviews and book chapters, searched conference proceedings and specialty journals, and contacted experts. STUDY SELECTION: We screened 1349 abstracts and excluded those that did not test in-home interventions or in which the mean age of the study population was younger than 70 years. After further exclusions, 17 articles describing 18 trials were analyzed. DATA EXTRACTION: Two reviewers independently screened abstracts. Discrepancies were resolved by consensus with a third reviewer. For each included trial, we extracted data on the study population and the characteristics of the intervention. Two of us extracted information on 3 end points: nursing home admissions, mortality, and functional status. One of us assessed trial quality, including an examination of the method of randomization, blinding of caregivers and research staff ascertaining outcomes, and proportion of patients included in analyses of the 3 end points. DATA SYNTHESIS: The 18 trials included 13 447 individuals aged 65 years and older. The effect on nursing home admissions depended on the number of visits performed during follow-up. The pooled relative risk (RR) was 0.66 (95% confidence interval [CI], 0.48-0.92) for trials in the upper tertile (>9 visits) but was 1.05 (95% CI, 0.85-1.30) in the lower tertile (0-4 visits). Functional decline was reduced in trials that used multidimensional assessment with follow-up (RR, 0.76; 95% CI, 0.64-0.91) but not in other trials (RR, 1.01; 95% CI, 0.92-1.11). Functional decline was reduced (RR, 0.78; 95% CI, 0.64-0.95) in trials with a control group mortality rate in the lower tertile (3.4%-5.8%) but not (RR, 0.98; 95% CI, 0.84-1.13) in those with a control-group mortality rate in the upper tertile (8.3%-10.7%). A beneficial effect on mortality was evident in younger study populations (RR, 0.76; 95% CI, 0.65-0.88 for ages 72.7-77.5 years) but not in older study populations (RR, 1.09; 95% CI, 0.92-1.28 for ages 80.2-81.6 years). CONCLUSION: Preventive home visitation programs appear to be effective, provided the interventions are based on multidimensional geriatric assessment and include multiple follow-up home visits and target persons at lower risk for death. Benefits on survival were seen in young-old rather than old-old populations. PMID- 11866652 TI - A 44-year-old woman with borderline personality disorder. PMID- 11866653 TI - A 45-year-old woman with obsessive-compulsive disorder, 1 year later. PMID- 11866654 TI - Ethics of preimplantation diagnosis for a woman destined to develop early-onset Alzheimer disease. PMID- 11866655 TI - Contact investigations and the continued commitment to control tuberculosis. PMID- 11866656 TI - The burden of out-of-pocket payments for health care in Tbilisi, Republic of Georgia. AB - CONTEXT: In the 1990s, the Republic of Georgia instituted health care reforms to convert the centralized, state-operated health care system inherited from the Soviet Union to a decentralized, market-driven system of health care delivery. Under the new system, 87% of health care expenditures are financed through out-of pocket payments at the point of service. OBJECTIVE: To describe the effects of health care reforms on access to care and health care financing among ill residents of Tbilisi, Georgia. DESIGN, SETTING, AND PARTICIPANTS: A probability proportionate-to-size cluster survey conducted in 1999 of 248 households containing 306 household members who had been ill in the past 6 months in Tbilisi, Georgia. MAIN OUTCOME MEASURES: Reported health care utilization, out-of pocket expenditures, and financing practices. RESULTS: Of sick household members, 51% used official health care services at hospitals and clinics; 49% did not use official services and sought advice from relatives or friends, used traditional medicines, or did nothing. Those with serious illness were more likely to seek care through official services (82%) than those with nonserious illness (27%). Ninety-three percent of respondents said costs were the major deterrent to obtaining health care. Ten percent of ill household members reported that they were unable to obtain health care because of high costs; 16% reported being unable to afford all the medications necessary to treat their illness. Sixty-one percent of ill household members used savings to pay for health care expenditures and 19% of those able to obtain care had to use strategies such as borrowing money or selling personal items to pay for health care. Total out-of-pocket health care expenditures (53%) were paid for by borrowing money or selling personal items. A significant portion of households with ill members (87%) reported an interest in purchasing health care insurance. CONCLUSIONS: Economic disruption and health care reforms have led to access problems and out-of-pocket financing strategies that include reliance on personal savings, selling personal items, and borrowing money. Future reforms should consider an appropriate system for health care insurance risk pooling for the population of Tbilisi, Georgia. PMID- 11866662 TI - Oral contraceptives and the risk of thrombosis and atherosclerosis. AB - Oral contraceptives containing synthetic oestrogens have been used successfully as birth control for > 40 years and are currently prescribed to > 100 million women worldwide. Several new progestins have been introduced and the third generation of progestins has now been available for two decades. Oral contraceptives are prescribed over a prolonged period of time and therefore substantially impact on hormonal, metabolic and plasmatic functions. Oral contraceptives increase the risk for venous thrombosis and pulmonary embolism, particularly if associated with confounding factors, such as genetic predisposition, smoking, hypertension or obesity. The risk of developing coronary artery disease is also increased in users with cardiovascular risk factors. This article discusses mechanistic and clinical issues and reviews the need for novel approaches targeting the considerable side effects in order to reduce cardiovascular morbidity in women using oral contraceptives. PMID- 11866663 TI - Advances in male contraception. AB - Advances in contraception have been almost exclusively female-directed despite the widespread use of male methods worldwide and increasing calls for the burden of contraception to be more evenly shared. Of the several potential approaches to novel male methods, the hormonal approach is the nearest to fruition. The use of testosterone as a reversible contraceptive agent in men has been demonstrated in studies undertaken by the WHO over the last decade. However, an agent that results in universal azoospermia without significant side effects remains elusive. Consequently, combination approaches with progestogens, anti-androgens, 5 alpha-reductase inhibitors and gonadotrophin-releasing hormone (GnRH) antagonists have been evaluated with the aim of improving contraceptive efficacy. Different methods of androgen delivery are also being developed in order to minimise extra-testicular effects and improve acceptability. This review will focus on efforts to develop a safe, acceptable, efficacious hormonal contraceptive for men. PMID- 11866664 TI - Therapeutic potential of CPT I inhibitors: cardiac gene transcription as a target. AB - Inhibitors of carnitine palmitoyl-transferase I (CPT I), the key enzyme for the transport of long-chain acyl-coenzyme A (acyl-CoA) compounds into mitochondria, have been developed as agents for treating diabetes mellitus Type 2. Findings that the CPT I inhibitor, etomoxir, has effects on overloaded heart muscle, which are associated with an improved function, were unexpected and can be attributed to selective changes in the dysregulated gene expression of hypertrophied cardiomyocytes. Also, the first clinical trial with etomoxir in patients with heart failure showed that etomoxir improved the clinical status and several parameters of heart function. In view of the action of etomoxir on gene expression, putative molecular mechanisms involved in an increased expression of SERCA2, the Ca(2+) pump of sarcoplasmic reticulum (SR) and alpha-myosin heavy chain (MHC) of failing overloaded heart muscle are described. The first 225 bp of human, rabbit, rat and mouse SERCA2 promoter sequence have high identity. Various cis-regularory elements are also given for the promoter of the rat cardiac alpha MHC gene. It is hypothesised that etomoxir increases glucose-phosphate intermediates resulting in activation of signalling pathway(s) mediated by phosphatases. Regarding the possible direct action of etomoxir on peroxisome proliferator activated receptor alpha (PPAR-alpha) activation, it could upregulate the expression of various enzymes that participate in beta-oxidation, thereby modulating some effects of CPT 1 inhibition. Any development of alternative drugs requires a better understanding of the signal pathways involved in the altered gene expression. In particular, signals need to be identified which are altered in overloaded hearts and can selectively be re-activated by etomoxir. PMID- 11866665 TI - Anti-androgens for the treatment of hirsutism. AB - Many alternatives exist for treating hirsutism. Based on an analysis of scientific literature and on the experiences of the author, the most common anti androgen agents are discussed in this review. Androgen receptor blockers (cyproterone acetate, flutamide and spironolactone), 5 alpha-reductase inhibitors (finasteride) and androgen-suppressing agents (gonadotrophin-releasing hormone [GnRH] agonists, oestroprogestins, corticosteroids and insulin-sensitising agents) are evaluated and compared. The importance of diagnosis in choosing the most appropriate anti-androgen treatment is also discussed. PMID- 11866666 TI - New developments in the treatment of inflammatory bowel disease. AB - Therapy of inflammatory bowel disease (IBD) is rapidly changing with the advent of new discoveries in disease pathogenesis. The need for targeted therapies against the uncontrolled immuno-inflammatory reaction in IBD together with a prerequisite for minimal side effects is driving improvement in old medicines and is leading to the development of new drugs. This review introduces emerging changes in IBD treatment, such as improvements in conventional IBD medications or their use. Balsalazide, budesonide and changes in the use of 5-aminosalicylate (5 ASA) products and purine analogues, such as azathioprine, are discussed. Additionally, studies examining the role of drugs newly introduced into IBD therapy, such as mycophenolate mofetil (MMF), thalidomide and heparin, are stated. Emerging biological therapies, such as therapies against TNF, therapies to enhance anti-inflammatory cytokines, therapeutic manoeuvres to disrupt immune cell trafficking, anti-oxidant therapies, as well as non-conventional treatments, such as diet therapies, prebiotics and probiotics, and helminth therapies are discussed. PMID- 11866667 TI - HIV-1 protease inhibitors in development. AB - Several pharmaceutical companies have developed an increasing number of second generation protease inhibitors (PI) during the last few years. Many of these compounds have been in preclinical trials and some are now in clinical use. All drugs in this category have been designed to be well absorbed and overcome the crucial problem of cross-resistance within this class of compounds. Taking into account the rapid occurrence of PI cross-resistance, clinicians who are treating patients with the HIV-1 infection will need new active PIs in the near future. The clinical and antiviral efficacy of the new molecules versus the older PIs will be investigated through comparative trials that are likely to be completed over the next 12 months. These third-generation PIs currently in development will be the subject of our review. PMID- 11866668 TI - Fondaparinux: a synthetic heparin pentasaccharide as a new antithrombotic agent. AB - Fondaparinux (Arixtra, Sanofi-Synthelabo/Organon) is the first of a new class of antithrombotic agents distinct from low molecular weight heparins (LMWHs) and heparin. It is a chemically synthetic pentasaccharide mimicking the site of heparin that binds to antithrombin III (AT). It exhibits only factor (F) Xa (FXa) inhibitor activity via binding to AT, which in turn inhibits thrombin generation. In contrast to heparin and LMWH, plasma anti-Xa activity corresponds directly to levels of fondaparinux. It does not release tissue factor pathway inhibitor (TFPI). There is nearly complete bioavailability by the sc. route, rapid onset of action, a prolonged half-life in both iv. and sc. (14 - 20 h) dosing regimens and no metabolism preceding renal excretion. Phase IIb clinical studies have identified a dose of 2.5 mg once-daily for prophylaxis of venous thrombosis. Four Phase III studies (n > 7000) have demonstrated a combined 50% relative risk reduction of venous thromboembolic events in orthopaedic surgery patients in comparison to the LMWH, enoxaparin. Haemmorrhagic complications for fondaparinux were either comparable to or higher than those for LMWH. The activated partial thromboplastin time (aPTT) is not affected by fondaparinux. At present, laboratory monitoring is not recommended. Clinical trials for treatment of established thrombosis, coronary syndromes and adjunct to thrombolytic therapy are in progress. PMID- 11866669 TI - Review of the safety and efficacy of voriconazole. AB - Voriconazole is a new triazole antifungal agent structurally related to fluconazole, but with improved potency and spectrum of activity. Voriconazole has good in vitro activity against Candida species, Cryptococcus neoformans, Aspergillus spp. and other mould spp. Initial clinical studies and case reports demonstrate efficacy with voriconazole against invasive aspergillosis and infections caused by C. neoformans, Scedosporium apiospermum, Blastomyces dermatitidis, Coccidioides immitis and Histoplasma capsulatum. Voriconazole is available both as oral and iv. preparations and exhibits complex pharmacokinetics. This drug is metabolised by the cytochrome (CYP) P450 enzyme system and therefore, has potential drug interactions. This review evaluates the current literature regarding the safety and efficacy of voriconazole. PMID- 11866670 TI - Endothelin receptor antagonists in cardiology clinical trials. AB - Endothelin-1 (ET-1) is enhanced and has been demonstrated to be a prognostic marker in patients with advanced stages of heart failure, acute ischaemic syndromes, myocardial infarction and pulmonary hypertension. Activation of the endothelin (ET) system is associated with adverse haemodynamic consequences in patients with congestive heart failure and results in coronary vasoconstriction in patients with coronary artery disease (CAD). Moreover, ET-1 raises blood pressure, induces vascular and myocardial hypertrophy and acts as the natural counterpart of nitric oxide (NO), which exerts vasodilating, antithrombotic and antiproliferative effects. This article reviews recently completed and ongoing clinical trials examining the effects of ET receptor antagonists in patients with heart failure, CAD, arterial hypertension and pulmonary hypertension. PMID- 11866671 TI - Lopinavir/ritonavir (ABT-378/r). AB - Despite major advances in HIV research, eradication of HIV from the body is not yet possible. However, current antiretroviral (ARV) therapy can achieve disease control via viral suppression below the limits of detection of currently available assays. This has led to a marked decline in morbidity and mortality associated with the development of opportunistic infections and malignancies. Since viral suppression appears to be the most achievable goal of current therapy, there arises a need for new and more potent ARV agents in order to maintain viral suppression. Many of the currently available protease inhibitors (PIs) have a high protein-binding ability, short plasma half-life [1] and pharmacokinetic interactions with food or other drugs [2]. This can result in sub optimal plasma drug concentrations, which may allow the virus to break through and replicate, leading to the development of resistant mutants [1]. Lopinavir/ritonavir (LPV/r; Kaletra, Abbott Laboratories) is a new PI consisting of a co-formulation of lopinavir and low-dose ritonavir. The sub-therapeutic dose of ritonavir (a potent cytochrome P450 [CYP] 3A4 inhibitor) inhibits the metabolism of lopinavir, resulting in higher lopinavir concentrations than when lopinavir is administered alone [3]. This pharmacokinetic interaction is associated with a high lopinavir trough level:wild type median effective concentration (EC(50)) ratio and good general tolerability when compared with other currently licensed PIs [4]. The concept of pharmacokinetic enhancement - boosting - is not new as ritonavir has previously been utilised in this context with other PIs. The relationship between plasma and intracellular drug levels has yet to be clarified. What has been ascertained from pharmacokinetic studies thus far is the correlation between ARV trough plasma concentrations (C(min)) and virological outcome [5,6]. LPV/r exemplifies how the pharmacokinetic profile of a drug can be modified to attain sufficient C(min) to suppress pheno- and genotypically resistant viral strains, as well as provide a pharmacological barrier to the emergence of resistance [7]. LPV/r reduces pill-burden and aids compliance, as shown by encouraging results in the treatment of both ARV-naive and -experienced patients. PMID- 11866672 TI - The economic impact of chronic obstructive pulmonary disease. AB - The cost burden associated with chronic bronchitis and emphysema, collectively known as chronic obstructive pulmonary disease (COPD), is large. The disease impacts not only on patients but caregivers and society as well. An estimated 16 million people in the US are currently diagnosed with COPD, the majority having chronic bronchitis. Mortality associated with this disease is on the upswing, as is its prevalence in the female population and the elderly. It is currently the fourth most common cause of death both in the US and worldwide. To date, the only proven cost-effective therapies for the disease are the cessation or prevention of smoking, which is the single most common cause of COPD, and vaccination to prevent influenza and pneumococcal infection. Hospitalisation and associated costs represent the greatest healthcare expenditures for people with the disease. Long-term oxygen therapy is also among the most costly interventions in terms of total money spent on direct medical costs for COPD treatment, although it is probably cost-effective because of its positive impact on rates of mortality. In fact, oxygen therapy is the only intervention to date that has been shown to decrease death rates due to COPD. Appropriate treatment with medication has the potential to decrease resource utilisation but does not appear to affect death rates. Similarly, pulmonary rehabilitation programs appear to benefit patients in terms of quality of life; however, long-term cost-effectiveness and effects on mortality have yet to be elucidated. Indirect costs also contribute a substantial part of the economic burden of the disease but are significantly harder to quantify. PMID- 11866673 TI - Pharmacoeconomics of opioid therapy for chronic non-malignant pain. AB - Chronic non-malignant pain (CNMP) is widely prevalent and associated with significant costs. Costs related to chronic pain include medical services and medications, treatment of medication-related toxicity and work absenteeism. The use of non-narcotic analgesics is associated with inadequate pain-relief for many patients, as well as significant and costly organ toxicity. When used appropriately and judiciously, opioid medications can be a useful addition to the treatment plan for patients with CNMP. Opioids can provide long-term, safe and cost-effective pain relief. PMID- 11866674 TI - Pharmacoeconomic evidence and considerations for triptan treatment of migraine. AB - The 5-hydroxytriptamine (5-HT)(1B/1D) agonists (i.e., the triptans) are highly effective in migraine but their high cost relative to other treatments necessitates careful analysis of their cost-effectiveness. The majority of pharmacoeconomic evaluations, nearly all of which deal with sumatriptan, indicate that from the societal perspective triptans are dominant: health outcomes are improved while the overall cost of migraine is reduced. However, the results are more ambiguous from the perspective of the health care payer: reductions in non drug medical costs are unlikely to offset fully the high drug cost. Thus far, few analyses have explicitly included quality of life (QOL) in their analyses and pharmacoeconomic analyses comparing the different triptans are scant. More research is required in these areas. PMID- 11866675 TI - Pharmacoeconomics of drug therapy for atopic dermatitis. AB - Atopic dermatitis is an increasingly prevalent common childhood disease. While the majority of patients have mild disease, atopic dermatitis can cause considerable distress to patients and their caregivers, with significant social and financial cost to families. With a prevalence of 15 - 20% in Western countries, atopic dermatitis also has a considerable health and societal cost to the community. Many new treatments have been shown to be therapeutically effective, particularly in severe disease, including cyclosporin A (Neoral, Novartis AG), interferon, tacrolimus (Fujisawa Pharmaceutical Co. Ltd.) and iv. immunoglobulin. These are expensive when compared to standard treatments like emollients and topical corticosteroids and have significant adverse effects that limit their use. Additional costs related to monitoring are incurred and the long term safety of these treatments is yet to be determined. However, an advantage over more traditional therapies is their ability to produce benefits even after treatment ceases. Treatments that produce long-term remissions have a greater likelihood of being cost-effective. With monetary constraints on healthcare and the importance governments place on reducing drug costs, economic evaluations are becoming an increasingly important factor for drug acceptance. Those evaluating cost-effectiveness should pay particular attention to the potential reduction in indirect and intangible costs. Unfortunately, there is a dearth of cost effectiveness studies in atopic eczema and this needs to be addressed with some urgency. PMID- 11866677 TI - Gastrointestinal side effects of NSAIDs - pharmacoeconomic implications. AB - NSAIDs are frequently used as analgesics. They have a high incidence of GI adverse effects that have both social and economic costs. These costs impose a considerable strain on healthcare resources. This review discusses the epidemiology and economic cost of these adverse GI events. It also highlights the variability in risk and the impact of newer drugs. Risk stratification and the subsequent guided use of NSAIDs is likely to limit the pharmacoeconomic implications. PMID- 11866676 TI - Economic aspects of medical erectile dysfunction therapies. AB - The introduction of safe and effective oral therapies has changed the significance of erectile dysfunction (ED) forever. Seldom has a new drug changed the landscape of pharmacotherapy in so many ways as sildenafil (Viagra, Pfizer). This article highlights the various oral, intracavernosal and intraurethral therapies currently available for ED and evaluates the socio-economic impact of each method. PMID- 11866678 TI - Cost-effectiveness of antibiotic prophylaxis for bacterial arthritis. AB - The outcome of bacterial arthritis is generally poor: the mortality is 10 - 15% and there is loss of joint function in 25 - 50% of the survivors. The incidence of bacterial arthritis is low: 2 - 6 cases per 100,000 people per year. Risk factors are age, joint disease (especially rheumatoid arthritis [RA]), diabetes mellitus and the presence of a prosthetic joint. The predominant situations that can lead to bacterial arthritis are skin infections of the feet and rarely invasive medical or dental procedures. Due to the severity of the disease, antibiotic prophylaxis of haematogenous bacterial arthritis in patients with prosthetic joints is advocated. However, due to the rarity of the disease it is unclear whether the advantages of prophylaxis outweigh the disadvantages of the large-scale use of antibiotics, such as side effects, costs and bacterial resistance. In a decision-analysis of a large group of patients with joint diseases, antibiotic treatment of skin infections appeared to be cost-effective in the prevention of haematogenous bacterial arthritis, mainly in high-risk patients. On the other hand, prophylaxis around medical or dental procedures was not cost-effective, except possibly in a small group of patients with increased risk. PMID- 11866679 TI - Ketolides in the treatment of respiratory infections. AB - The ketolides are a new class of macrolides specifically designed to combat respiratory tract pathogens that have acquired resistance to macrolides. The ketolides are semi-synthetic derivatives of the 14-membered macrolide erythromycin A. There are currently two ketolides in the late stages of clinical development in the US (telithromycin [HMR-364, Kelek; Aventis] and ABT-773 [Abbot Laboratories]), as well as newer compounds in earlier stages of testing. Ketolides have a mechanism of action very similar to that of erythromycin A. They potently inhibit protein synthesis by interacting close to the peptidyl transferase site of the bacterial 50S ribosomal subunit. Ketolides bind to ribosomes with higher affinity than macrolides. The ketolides exhibit good activity against Gram-positive and some Gram-negative aerobes and have are active against macrolide-resistant Streptococcus species, including most mef A and erm B strains of Streptococcus pneumoniae. Ketolides have pharmacokinetics which allow once-daily dosing and extensive tissue distribution with very high uptake into respiratory tissues and fluids relative to serum. Evidence suggests the ketolides are primarily metabolised by the cytochrome P450 (CYP) enzyme system in the liver and that elimination is a combination of biliary, hepatic and urinary excretion. Clinical trial data are only available for telithromycin and have focused on respiratory tract infections (RTIs) including community-acquired pneumonia (CAP), acute exacerbations of chronic bronchitis (AECB), sinusitis and streptococcal pharyngitis. Bacteriological and clinical cure rates have been similar to comparators. Ketolides have similar safety profiles to the newer macrolides. In summary, early clinical trials support the clinical efficacy of the ketolides in common RTIs, including activity against macrolide-resistant pathogens. PMID- 11866680 TI - The use of retinoids in the prevention and treatment of skin cancer. AB - There has been a significant increase in the number of cases of skin cancer diagnosed in the US in the past few years. Thus, it seems appropriate to review the available compounds that might be used in the chemoprevention of these lesions. This review focuses on the retinoids and details their results in clinical trials for treatment and prevention of skin cancer. Side effects of the various retinoid derivatives are also discussed. It is concluded that isotretinoin (13-cis-retinoic acid) is the most effective retinoid for the prevention of non-melanoma skin cancers in high-risk patients in clinical trials. Current basic research is focused on developing receptor-selective retinoids which would have a higher therapeutic index for the treatment and chemoprevention of skin cancer. PMID- 11866681 TI - Pharmacological treatments for rhinosinusitis. AB - Acute bacterial rhinosinusitis (ABRS) is a well-recognised and common problem confronting many primary care physicians but the abuse of antibiotic therapy for viral aetiologies of ABRS has lead to widespread bacterial resistance. The once easily-eradicated pathogens have developed many mechanisms to resist antimicrobial therapies. The most common pathogens Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis are still found in cultures of sinus cavities but Staphylococcus and possibly some anaerobes play an important role in the development of the chronic stage of rhinosinusitis. The choice of antibiotic therapy for both ABRS and chronic rhinosinusitis (CRS) are reviewed, along with considerations for ancillary therapy. Note is made of the role of chemical mediators in the inflammatory stage possible strategies to reduce this problem. Allergic fungal rhinosinusitis results from a hypersensitivity reaction to demetiaceous fungi. Cure of the effects of rhinosinusitis depends on the establishment of a healthy sinus cycle. PMID- 11866682 TI - Lamivudine for the treatment of chronic hepatitis B. AB - Lamivudine (Zeffix, Epivir, GlaxoSmithKline) is the most important recent advance in the treatment of chronic hepatitis B in both adults and children. It is the only available oral treatment and has an excellent safety profile, which makes it even more attractive. It increases the rate of hepatitis B e antigen (HBeAg) loss and seroconversion in compensated chronic HBeAg-positive carriers, with subsequent improvement of histology at a similar rate as IFN-alpha. Lamivudine is mostly active in patients with elevated transaminases and is not effective in compensated patients with quiescent disease. Long-term follow-up studies are still required to evaluate long-term benefits, including those on hepatitis B surface antigen (HBsAg) seroconversion rate and disease evolution control. In decompensated patients, the drug can stabilise and improve liver function, allowing the patient to wait safely for transplantation. Patients may improve to such an extent that transplantation can be postponed. Combined with hepatitis B immunoglobulin (HBIG), lamivudine considerably decreases the risk of graft re infection after transplantation. It is also active in chronic HBeAg-negative hepatitis patients, for whom IFN is less efficient. The major drawback is the emergence of the tyrosine-methionine-aspartate-aspartate (YMDD) mutation, which prevents further efficacy of the drug and may lead to flares of hepatitis. Due to the questions the YMDD mutation raises and because hepatitis B is a complex disease, indications for treatment must be established with care and only by physicians with expert knowledge of the disease, the drug and YMDD mutation related issues. PMID- 11866683 TI - Seretide for obstructive lung disease. AB - Seretide (Advair [North America], GlaxoSmithKline) is an inhaler combination formulation intended for the maintenance therapy of obstructive airways disease. Seretide was developed and made available initially as three multi-dose, dry powder inhaler formulations delivering 50 microg/puff of the long acting beta(2) agonist salmeterol and either 100, 250 or 500 microg/puff of the inhaled corticosteroid fluticasone propionate. In addition to the initial multi-dose dry powder inhaler system (Diskus or Accuhaler), a chlorofluorocarbon (CFC)-free pressurised aerosol formulation has become available. Studied mostly extensively as a maintenance therapy for patients with persistent asthma, the combination inhaler is at least equivalent to its components administered separately and is superior to monotherapy with salmeterol or inhaled corticosteroid in both paediatric and adult populations. The combination has a logical role in the treatment of moderate-to-severe asthma, offering the advantage of increased convenience and possibly improved compliance. In addition to improvements in lung function, symptom scores and quality of life, the combination therapy reduces exacerbation rates, an outcome that contributes to favourable cost-effectiveness. A role as initial maintenance therapy in all forms of persistent asthma is also plausible but there are fewer data concerning the impact of Seretide in milder forms of persistent asthma. Clinical trials are underway to examine the potential role of Seretide in patients with chronic obstructive pulmonary disease (COPD). Salmeterol has been shown to be an effective first-line bronchodilator in COPD and fluticasone has been shown to reduce the frequency and or severity of exacerbations in COPD patients in two key trials. At a time when the prevalence of both asthma and COPD is increasing, Seretide is a valuable step in the management of these common obstructive lung diseases. PMID- 11866684 TI - Clopidogrel: a CURE in acute coronary syndromes? AB - The standard approach to preventing acute coronary syndromes (ACSs)has been to inhibit platelet aggregation with aspirin and to inhibit blood coagulation with low molecular-weight heparin (LMWH). Even with this combination there is still a substantial short and long-term cardiovascular risk. The Clopidogrel in Unstable angina to prevent Recurrent Events (CURE) trial [1] compared clopidogrel plus aspirin against aspirin alone in patients with ACSs. The clopidogrel regimen was a loading dose of 300 mg p.o. followed by 75 mg/day and the recommended dose of aspirin was 75 - 325 mg/day. The first primary outcome was a composite of death from cardiovascular causes, non-fatal myocardial infarction (MI) or stroke and this occurred significantly less often in the clopidogrel than the placebo group (9.3 vs. 11.4%). Although there were more clopidogrel patients with life threatening bleeding (clopidogrel 2.2%, placebo 1.8%), this represented GI haemorrhages and bleeding at sites of arterial puncture rather than fatal bleeding. This trial suggests a role for clopidogrel in the long-term treatment of ACSs. PMID- 11866685 TI - Future developments in assisted reproduction in humans. AB - The advent of human in vitro fertilization (IVF) over 30 years ago has made the oocyte and preimplantation embryo uniquely accessible. This accessibility has given rise to new micromanipulation techniques, such as intracytoplasmic sperm injection for treatment of male infertility, as well as embryo biopsy for preimplantation diagnosis of both genetic disease and aneuploidy, a major cause of early embryo demise and miscarriage. In the UK, average pregnancy rates after IVF and embryo transfer are < 25%, even after transfer of several embryos. Unfortunately, a third of these pregnancies involve multiple gestations. Research is currently focusing on methods to improve IVF success rates while reducing twin and triplet pregnancies and their associated increased morbidity and mortality. One approach is to develop screening methods to identify the most viable embryos, so that transfer of fewer healthy embryos will result in a higher proportion of singleton pregnancies. Screening methods include optimizing culture conditions for prolonged culture and selection of viable blastocysts for transfer, or embryo biopsy and aneuploidy screening. Assisted reproduction is also increasingly important in other branches of medicine: survival rates for cancer sufferers are improving continually and there is now a significant need for approaches to preserve fertility after sterilizing chemo-and radiotherapy treatment. Techniques for cryopreserving male and female gametes or gonadal tissue are being developed, although systems to grow and mature these gametes are in their infancy. Finally, there are also concerns regarding the safety of these new assisted reproductive technologies. PMID- 11866686 TI - The earliest stages of folliculogenesis in vitro. AB - In recent years several follicle culture systems have been pioneered in different mammalian species for studying ovarian folliculogenesis and culturing immature oocytes. Applications of these in vitro techniques include fertility preservation for humans, conservation of rare animals and development of oocyte banks for research purposes. Immature female gametes in the ovarian cortex can be cryopreserved for later use if culture techniques are available afterwards to promote growth and maturation. This review focuses on biochemical and biophysical factors related to oocyte culture in mice, the only animal in which live offspring have been produced after folliculogenesis in vitro. The advantage of using mice for these studies is that, in parallel to development of follicle culture systems, essential knowledge on folliculogenesis can be obtained from knockout mouse models. Recent experiments in mice stressed the principal role of the oocyte in follicle development and the strict timing of the biological processes underlying oogenesis in vitro. In large domestic animals and humans, study of oocyte culture is confounded by the constitutively prolonged nature of ovarian follicle development. In humans, only some aspects of follicle development have been studied because of the limited availability of suitable material for experimentation, technical difficulties related to manipulation of very small structures and lack of knowledge on physiological regulation of the early stages of follicle growth. Only a few reports describe ovarian follicular growth in vitro. In this review, relevant information on hormonal and growth factor regulation of the earliest stages of follicle growth in mammals is reviewed. Techniques are becoming available for the precise isolation of distinct classes of follicle and powerful molecular biology techniques can be used in studies of ovarian tissue culture. PMID- 11866687 TI - Voltage-operated calcium channels in male germ cells. AB - The acrosome reaction is a key event in fertilization. Current models for induction of the acrosome reaction incorporate a necessary influx of Ca(2+), which is mediated by agonist-induced gating of ion channels in the sperm plasma membrane. The difficulty of applying electrophysiological techniques to spermatozoa has severely hampered studies on the expression of functional ion channels in these cells. However, during the last few years, a combination of molecular and physiological techniques (applied to immature spermatogenic cells) has elucidated both the expression of Ca(2+) channels in male germ cells and their role in induction of the acrosome reaction. It now appears that a range of voltage-operated Ca(2+) channels, similar to those that occur in somatic cells, is expressed in spermatozoa. Male rodent germ cells express a low-voltage activated (T-type) channel that is regulated by membrane potential and provides the primary Ca(2+) influx mechanism in zona pellucida-stimulated spermatozoa. In human spermatozoa, similar channels are apparently expressed, but their function in induction of the acrosome reaction has yet to be established. A range of other, high voltage-activated channels also appear to be present in rodent and human spermatozoa, but their roles are not yet known. In this review, the structure and characteristics of voltage-operated Ca(2+) channels are outlined and the evidence for their expression and function in male germ cells is assembled and discussed. PMID- 11866688 TI - Peritoneal environment, cytokines and angiogenesis in the pathophysiology of endometriosis. AB - Endometriosis, defined by the presence of viable endometrial tissue outside the uterine cavity, is a common condition affecting 2-3% of women of reproductive age. Today, a composite theory of retrograde menstruation with implantation of endometrial fragments in conjunction with peritoneal factors to stimulate cell growth is the most widely accepted explanation. There is substantial evidence that immunological factors and angiogenesis play a decisive role in the pathogenesis of endometriosis. In women with endometriosis, there appears to be an alteration in the function of peritoneal macrophages, natural killer cells and lymphocytes. Furthermore, growth factors and inflammatory mediators in the peritoneal fluid, produced mainly by peritoneal macrophages, are altered in endometriosis, indicating a role for these immune cells and mediators in the pathogenesis of this disease. PMID- 11866689 TI - Expression of a novel reticulon-like gene in human testis. AB - Identification of genes that are specifically expressed in the adult testis or the fetal testis is important for the study of genes related to the development of the testis. In this study, a human testis cDNA microarray was established. PCR products of 9216 clones from a human testis cDNA library were dotted on a nylon membrane; mRNA from adult and fetal testes were purified and probes were prepared by a reverse transcription reaction with testis mRNA as template. The microarray was hybridized with probes of adult and fetal testes, and 96.8 and 95.4% of clones were positive, respectively. In total, 731 clones were differentially expressed: 592 were highly expressed in adult testis and 139 were highly expressed in fetal testis. Among these genes, a new reticulon (Rtn)-like gene was detected and named Rtn-T. Rtn-T was highly expressed in adult human testis. The cDNA of Rtn-T contains 3491 bp and the putative protein had 968 amino acids. This protein is homologous to the six known members of the Rtn family (KIAA0886, Rtn xL, reticulon 4a, Nogo-A, Nogo-A short form, and brain my043) but was different at the 5' end. All homologues originate from one gene, and result from both different promotor regions and different splicing. Rtn-T lacks the first exon and contains a second exon that is lacking in the other homologues. Rtn-T is shorter than KIAA0886, Rtn xL, reticulon 4a and Nogo-A, but longer than the Nogo-A short form and brain my043. Sequence analysis showed that Rtn-T protein has two hydrophobic regions that may be membrane-spanning domains. Expression profiles showed that Rtn-T is specifically and strongly expressed in testis. The results of the present study indicate that the Rtn-T gene is differentially expressed in adult and fetal testes and encodes a membrane protein that may have a function in testis development. PMID- 11866690 TI - Oestrogenic activity of the hop phyto-oestrogen, 8-prenylnaringenin. AB - The female flowers of the hop plant (hop cones) are used as a preservative and as a flavouring agent in beer. A novel phyto-oestrogen, 8-prenylnaringenin, was recently identified in hops and this study was undertaken to characterize the oestrogenic activity of this compound using a combination of in vitro and in vivo assays. Natural and semi-synthetic 8-prenylnaringenin showed similar bioactivities both in a yeast screen transfected with the human oestrogen receptor and in oestrogen-responsive human Ishikawa Var-I cells. 8 Prenylnaringenin showed comparable binding activity to both oestrogen receptor isoforms (ER alpha and ER beta). 8-Prenylnaringenin extracted from hops contains similar amounts of both (R)- and (S)- enantiomers, indicating that the compound is normally formed non-enzymatically. Both enantiomers showed similar bioactivity in vitro and similar binding characteristics to ER alpha and ER beta. The oestrogenic activity of 8-prenyl-naringenin in vitro was greater than that of established phyto-oestrogens such as coumestrol, genistein and daidzein. The high oestrogenic activity was confirmed in an acute in vivo test using uterine vascular permeability as an end point. When the compound was given to ovariectomized mice in their drinking water, oestrogenic stimulation of the vaginal epithelium required concentrations of 100 mug ml(-1) (about 500-fold greater than can be found in any beer). PMID- 11866691 TI - Hormonal regulation of apoptosis in rabbit granulosa cells in vitro: evaluation by flow cytometric detection of plasma membrane phosphatidylserine externalization. AB - Annexin V and propidium iodide bivariate analysis and the TUNEL method were used to quantify hormonal regulation of apoptosis in rabbit granulosa cells from preovulatory follicles in vitro. The aim of this study was to analyse comparatively the effects of gonadotrophins and their second messenger in the regulation of granulosa cell apoptosis in (i) cultured isolated granulosa cells and (ii) granulosa cells scraped from cultured follicles. The results showed that increasing doses of FSH had no effect on apoptosis of cultured isolated cells but caused a decrease in the number of apoptotic granulosa cells from preovulatory follicles cultured in serum-free conditions. Unlike FSH, addition of hCG did not modify apoptosis of granulosa cells significantly. In contrast, dibutyryl cAMP had an apoptotic effect in the two cellular models in the presence of serum. Moreover, a biphasic effect of dibutyryl cAMP in isolated granulosa cells was observed with an increase in the incorporation of [(3)H]thymidine into DNA at the lowest dose and an increase in apoptotic cell death at the highest dose. It was concluded that, in rabbits: (i) FSH requires follicle integrity to exert its anti apoptotic effect in granulosa cells; (ii) dibutyryl cAMP induces a dose-dependent apoptotic effect in granulosa cells cultured alone or obtained from cultured preovulatory follicles; and (iii) cAMP signals induce opposite effects on growth and apoptosis in granulosa cells. PMID- 11866692 TI - Use of energy substrates by various stage preimplantation pig embryos produced in vivo and in vitro. AB - The aim of in vitro embryo systems is to produce embryos of comparable quality to those derived in vivo. Comparison of embryo metabolism as an indicator of viability may be useful in optimization of culture conditions. The aim of the present study was to determine glucose, glutamine and pyruvate use by various stage pig embryos produced in vitro and in vivo. The results indicate that pig embryos use glucose via glycolysis in significant amounts at all stages examined, regardless of embryo origin. In vitro-derived embryos have significantly increased glycolytic activity after the eight-cell stage, whereas in vivo-derived embryos have increased glycolysis at the blastocyst stage. In vivo-derived embryos have higher rates of glycolysis compared with in vitro-derived embryos. Glucose usage through the Krebs cycle for in vitro- and in vivo-derived embryos increased significantly at the blastocyst stage. Pig embryos produced in vitro used constant amounts of glutamine throughout development, whereas in vivo derived embryos increased glutamine usage after the eight-cell stage. Pyruvate use was minimal at all stages examined for both in vitro- and in vivo-derived pig embryos, showing significant increases at the blastocyst stage. Krebs cycle metabolism of pyruvate, glutamine and glucose by in vivo-derived embryos was higher than that by in vitro-derived embryos. Current in vitro culture conditions produce pig embryos with altered metabolic activity, which may compromise embryo viability. PMID- 11866693 TI - Prostaglandin E(2) and F(2 alpha) production by equine conceptuses and concentrations in conceptus fluids and uterine flushings recovered from early pregnant and dioestrous mares. AB - A growing equine conceptus must suppress the cyclical release of PGF(2 alpha) from the endometrium to effect maternal recognition of its presence in the uterus. Paradoxically, the conceptus itself secretes PGF(2 alpha), together with other prostaglandins. In this study, the PGF(2 alpha) and PGE(2) content of, and production in vitro by, day 10-32 equine conceptuses were measured and the influence of pregnancy on the concentrations of these prostaglandins in the uterine lumen was examined. In vitro, the release of both prostaglandins per mg conceptus tissue was very high on day 10 after ovulation and lower thereafter. However, while PGF(2 alpha) production decreased further after day 18 of gestation, PGE(2) production remained high until day 32. Prostaglandin concentrations in yolk sac fluid were unaffected by gestational age and PGE(2) concentrations in this compartment were two to five times higher than PGF(2a) concentrations. PGF(2 alpha) concentrations reached high values in uterine flushings recovered from cyclic mares during days 14-16 after ovulation, the expected time of luteolysis, but were negligible in flushings recovered from pregnant mares at this time. Beyond day 18 of gestation, PGF(2 alpha) concentrations in uterine flushings were high and strikingly similar to those recorded during cyclical luteolysis. It is concluded that the equine conceptus effects maternal recognition of pregnancy primarily by inhibiting the ability of the endometrium to release PGF(2 alpha) during days 12-16 after ovulation. However, the conceptus appears to delay, rather than prevent, the development of the uterine PGF(2 alpha) release pathway and an alternative mechanism must prevent luteolysis from being triggered during days 18-32 of gestation. PMID- 11866694 TI - Role of FSH, numbers of FSH receptors and testosterone in the regulation of inhibin secretion during the seasonal testicular cycle of adult rams. AB - The regulation of inhibin secretion has not been elucidated fully in male ruminants. The aim of this study was to determine the relative importance of FSH and testosterone concentrations, and FSH receptors, in the control of secretion of immunoactive inhibin in rams. In Expt 1, temporal changes in hormone concentrations and testicular FSH binding were determined for two groups of rams (n = 4) kept under opposite, alternating 4 month periods of long (16 h light:8 h dark) and short (8 h light:16 h dark) days. Testicular biopsies (1-2 g) were collected when the testes were regressed, redeveloping, redeveloped and regressing. In Expt 2, separate groups of rams (n = 4) kept under natural photoperiod (latitude 45 degrees 48 minutes N) were designated as controls or passively immunized (for 3 weeks) with sufficient oestradiol antiserum to increase testosterone secretion without altering LH and FSH; this was done when the testes were regressed (non-breeding season) and redeveloped (breeding season). In both groups of rams (Expt 1), 'seasonal' increases in FSH concentrations began a few weeks earlier than did increases in inhibin concentrations. FSH reached maximum concentrations during testicular recrudescence, whereas numbers of FSH receptors in the testis and circulatory inhibin concentrations did not reach peak values until the testes were fully developed. Numbers of FSH receptors per testis, but not FSH concentration, were positively correlated (r = 0.65) with inhibin concentrations across the four stages of the testicular cycle. Near the end of testicular recrudescence early in the breeding season (Expt 2), relatively high FSH concentration was associated with increased abundance of FSH receptor mRNA (90%) and number of receptors (45%) in the testis and increased inhibin concentrations (50%), compared with when the testes were regressed. Moderate, physiological increases in testosterone secretion in immunized rams did not affect inhibin in either season. These results indicate that: (i) FSH stimulation of immunoactive inhibin secretion by Sertoli cells as testes recrudesce is via increases in secretion (early) and cognate receptors (late); (ii) FSH upregulates the synthesis of its own receptor late in recrudescence; and (iii) the positive correlation (r = 0.70) observed between circulatory testosterone and immunoactive inhibin does not reflect a causal relationship. PMID- 11866695 TI - Modulation of sex hormone secretion in cows by acute infection with bovine viral diarrhoea virus. AB - Bovine viral diarrhoea virus (BVDV) is a major pathogen of cattle and is responsible for considerable reproductive loss. In this study, the in vivo responses in six multiparous cows were investigated after a non-cytopathogenic BVDV challenge (strain Pe 515; 5 x 10(6) tissue culture infective dose 50) given 9 days before a synchronized ovulation. Six similar cows challenged with non infectious culture medium served as controls. The experimental noncytopathogenic BVDV infection was followed by a viraemia and leucopenia at days 5-9 after challenge, but no other clinical signs of infection were detected. However, the BVDV infection altered endocrine function. Mean LH pulse frequency immediately before CIDR withdrawal was lower (P < or = 0.05) in the BVDV-infected (2.17 +/- 0.34 pulses per 8 h) compared with the sham-infected (4.83 +/- 1.04 pulses per 8 h) animals. At day 3 after CIDR withdrawal, plasma oestradiol concentrations remained high (P < 0.05) in the infected cows (2.19 +/- 0.51 pg ml(-1)) compared with the sham-infected controls (0.72 +/- 0.29 pg ml(-1)). However, there was no difference in the peak oestradiol concentration (BVDV: 2.31 +/- 0.29 versus sham: 2.34 +/- 0.41 pg ml(-1)). In addition, non-cytopathogenic BVDV significantly (P < 0.05) increased the duration of the interval between ovulation and onset of the postovulatory progesterone increase (values 1.0 ng ml(-1)) (BVDV: 3.0 +/- 0.26 versus sham: 4.0 +/- 0.26 days). The viral infection also significantly (P < 0.01) decreased mean plasma progesterone concentrations between day 3 and day 11 after ovulation (BVDV: 2.59 +/- 0.32 versus sham: 4.13 +/- 0.27 ng ml(-1)). These data show that non-cytopathogenic BVDV viraemias during the follicular phase can modulate the secretion of gonadotrophins and sex steroids, in particular progesterone, during a synchronized oestrous cycle. Therefore, viraemias during the follicular phase may reduce the fertility of cattle by disrupting the capacity of the ovulatory follicle to form a competent corpus luteum, thereby compromising early embryo development and maternal recognition of pregnancy. PMID- 11866696 TI - Modulatory effects of gonadotrophins and insulin-like growth factor on the secretion of inhibin A and progesterone by granulosa cells from chicken preovulatory (F1-F3) follicles. AB - The aim of this study was to compare the actions and interactions of gonadotrophins (LH and FSH) and an analogue of insulin-like growth factor I (LR3 IGF-I) on the secretion of inhibin A, inhibin B and progesterone by cultured chicken granulosa cells derived from the three largest (F1--F3) follicles of the preovulatory hierarchy. Treatment with LH or FSH promoted marked dose-(P < 0.0001) and time- (P < 0.0001) dependent increases in both inhibin A and progesterone secretion, with the magnitude of response (< 15-fold compared with basal) increasing over time in culture. Concentrations of inhibin B were below the detection limit in all samples. Initially, F1 cells were more LH-responsive than were F3 cells in terms of progesterone secretion (P < 0.02) but this difference between follicles decreased over time in culture. In contrast, LH induced inhibin A secretion tended to be highest from F3 cells, although this was not significant. Cells from F3 follicles were consistently more FSH-responsive than F1 cells in terms of both progesterone (P < 0.01) and inhibin A (P < 0.02) secretion. Initially, F1 cells were more responsive to LR3-IGF-I than were F3 cells in terms of progesterone secretion (P < 0.001) but were less responsive in terms of inhibin A secretion (P < 0.001). Again, these inter-follicle differences decreased over time in culture (not significant on day 3 of treatment). Co treatment experiments showed that LR3-IGF-I enhanced both LH- and FSH-induced secretion of inhibin A and progesterone in a time- (P < 0.001) and follicle- (P < 0.001) dependent way. Initially, F1 cells showed highest LR3-IGF-I enhancement of LH-induced inhibin A and progesterone secretion; in contrast, F3 cells showed the highest LR3- IGF-1 enhancement of FSH-induced inhibin A and progesterone secretion. These inter-follicle differences persisted over time in the case of FSH-induced hormone responses but not in the case of LH-induced responses, even though the relative degree of LR3-IGF-I enhancement increased markedly over time. Collectively, these data support a positive role for IGF-I, presumably of thecal origin, as an amplifier of gonadotrophin action on granulosa cell inhibin A and progesterone production by preovulatory chicken follicles. PMID- 11866697 TI - Induction of birth in the bandicoot (Isoodon macrourus) with prostaglandin and oxytocin. AB - As in eutherians, maturation of the fetal pituitary and adrenal glands together with an increase in prostaglandin and mesotocin or oxytocin production initiates birth in marsupials. In this study, prostaglandin (Lutalyse) or oxytocin (Syntocinon) were administered to pregnant bandicoots at 05:00 h on the calculated day of birth and the resultant effects were filmed for analysis. The administration of prostaglandin caused the bandicoot to adopt the birth position several minutes after injection (n = 2). However, the bandicoot did not give birth for several hours. Birth occurred at a similar time of day to that observed for untreated bandicoots (n = 7), between 08:00 h and 12:00 h. After an injection of oxytocin, the bandicoot assumed the birth position and birth occurred within several minutes. The young were alive while still connected to their allantoic stalks. However, they were unable to attach to the teats and did not survive (n = 4). The induced young were the colour of venous blood and died soon after the umbilicus was separated, indicating that the cardiopulmonary system of these neonates was underdeveloped and inadequate to maintain life. The results from this study demonstrate that prostaglandin is required to prepare the bandicoot for birth, and mesotocin is required for contraction of the uterus and for birth to occur. PMID- 11866698 TI - Effect of immunization of hamsters against recombinant P26h on fertility rates. AB - Despite the various contraceptive methods available, an effective and inexpensive method remains to be established. Immunocontraception may help to achieve this goal. P26h has been proposed as a candidate for the development of a male contraceptive vaccine. P26h, a hamster sperm protein, interacts with the zona pellucida. Furthermore, in vivo fertilization can be blocked completely by active immunization of male hamsters against P26h. Maltose binding protein (MBP)-P26 shares antigenic determinants with the native P26h present on cauda epididymal spermatozoa. The aim of the present study was to reproduce the immunocontraceptive properties of native P26h by immunizing male hamsters against a recombinant P26h fused with a maltose binding protein (MBP). Active immunization of male hamsters with the MBP-P26h showed that specific anti-P26h circulating IgGs could be generated. Mating of immunized male hamsters with superovulated females resulted in a significant decrease, 20-25%, in the fertilization rate. This result is in agreement with results from in vitro sperm zona pellucida binding assays. Indeed, the anti-recombinant P26h IgGs showed lower inhibitory properties when compared with anti-native P26h IgG. Despite the high anti-P26h IgG titres generated in hamsters, histological studies showed that active immunization has no pathological sequelae to the reproductive tissues. The potential of P26h as a candidate for a contraceptive vaccine is discussed. PMID- 11866699 TI - Importance of cooling rate and animal variability for boar sperm cryopreservation: insights from the cryomicroscope. AB - A series of experiments was set up to investigate the effect of different cooling rates on boar sperm cryosurvival using cryomicroscopy. The cooling protocols were split into two stages: (i) from +5 degrees C to -5 degrees C and (ii) from -5 degrees C to -50 degrees C. Fluorescent probes (SYBR14 and propidium iodide) were used to monitor plasma membrane integrity during the entire process. Cooling rates in the range 3 degrees C min(-1) to 12 degrees C min(-1) did not cause significant damage to the sperm plasma membrane between +5 degrees C and -5 degrees C; however, spermatozoa cooled at 24 degrees C min(-1) to -5 degrees C were slightly damaged. Motility was not particularly sensitive to variations in cooling rate. Cooling rates in the range 15 degrees C min(-1) to 60 degrees C min(-1) did not produce differences in sperm cryosurvival during freezing between -5 degrees C and -50 degrees C, or after thawing. In addition, cooling rates in the range 3 degrees C min(-1) to 80 degrees C min(-1) did not produce significant differences in sperm cryosurvival. However, slow freezing (3 degrees C min(-1)) induced a slight increase in the percentage of plasma membrane-damaged spermatozoa (propidium iodide-positive) at -50 degrees C. Inter-ejaculate and inter-boar differences in sperm cryosurvival were manifested independently of cooling rate. The sperm plasma membrane remained intact (SYBR14-positive) during cooling and freezing, but upon rewarming, the plasma membrane of a high proportion of spermatozoa was damaged (propidium iodide-positive), indicating that rewarming is a critical step of the freezing-thawing process. PMID- 11866700 TI - Quantitative assessment of testicular germ cell production and kinematic and morphometric parameters of ejaculated spermatozoa in the grey mouse lemur, Microcebus murinus. AB - Germ cell production and organization of the testicular epithelium in a prosimian species, the grey mouse lemur, Microcebus murinus, was investigated to extend knowledge of comparative primate spermatogenesis. In addition, semen samples collected from adult male lemurs (body weight 53-92 g; n = 16) by rectal probe electroejaculation were evaluated using computer-assisted morphometric and kinematic analysis of spermatozoa. Epididymidal spermatozoa were collected from six animals after hemicastration; the testes were weighed and prepared for stereological analysis and flow cytometry. The relative testis mass (as a percentage of body weight) ranged between 1.17 and 5.6%. Twelve stages of testicular seminiferous epithelium as described for macaques were applied and only a single stage was observed in most of the seminiferous tubule cross sections. On average (mean SD), a single testis contained 1870 +/- 829 x 10(6) germ cells and 35 +/- 12 x 10(6) Sertoli cells. Germ cell ratios (preleptotene:type B spermatogonia = 2, round spermatid:pachytene = 3; elongated spermatid:round spermatids = 1) indicated high spermatogenic efficacy. Sperm head dimensions and tail lengths of the ejaculated and epididymidal spermatozoa were similar. Percentages of defects (neck/mid-piece and tail) were low ( 10%) and similar for ejaculated and epididymidal spermatozoa. Spermatozoa were highly motile, characterized by extensive lateral head displacement, but relatively low progressive motility. In conclusion, the grey mouse lemur has unusually large testes with a highly efficient spermatogenic process and large sperm output. These features, together with the high proportion of morphologically normal and highly motile spermatozoa in the ejaculates, indicate that Microcebus murinus is a species in which sperm competition after ejaculation is likely to occur. The predominantly single spermatogenic stage system seems to be an ancestral feature among primates. PMID- 11866702 TI - Finding heroes, choosing good. PMID- 11866703 TI - An adolescent with acute lymphoblastic leukemia. PMID- 11866704 TI - Processes of change related to smoking behavior among cancer patients. AB - PURPOSE: The purpose of this study was to examine the degree to which transtheoretical model processes of change (methods and strategies for cessation) were associated with smoking status and quitting behavior (ie, intentions and attempts to quit) among patients with head and neck or lung cancer. The relationship between medical variables and processes of change was also explored. DESCRIPTION OF STUDY: Twenty-nine smokers and 45 abstainers who were recruited from treatment clinics within a comprehensive cancer center completed a brief survey. Multivariate analysis of variance and Pearson correlation procedures were used to evaluate hypothesized relationships. RESULTS: As hypothesized, quitters used behavioral processes such as counter-conditioning and reinforcement management significantly more than smokers and used self-reevaluation, an experiential process, significantly less than smokers. Contrary to the hypothesis, however, quit attempts and intentions were associated with both experiential (ie, consciousness raising and self-reevaluation) and behavioral (ie, reinforcement management and self-liberation) processes of change. Use of the processes of change was not influenced by medical variables, including cancer type, illness phase, disease stage, type of current medical treatment, and duration of illness. CLINICAL IMPLICATIONS: These findings suggest that behavioral counseling to promote smoking cessation for patients with cancer should involve assisting the patient to do the following: develop an awareness of the health risks related to continued smoking; devise and use alternative behaviors; implement the use of reinforcement strategies for cessation successes; and develop a sense of confidence and commitment about quitting as well as healthy lifestyle values. These strategies are discussed within the context of models and guidelines for smoking cessation in clinical practice. PMID- 11866705 TI - Community implementation of home care palliative care education. AB - PURPOSE: The purpose of this National Cancer Institute-funded training project was to improve end-of-life care agencies through implementation of a palliative care curriculum. The Home Care Outreach for Palliative Care Education (HOPE) program was designed to improve the knowledge and skills of home healthcare professionals who provide care to the terminally ill and their families at home. DESCRIPTION OF PROGRAM: A five-module palliative care education curriculum was designed and implemented in five home care agencies in the Los Angeles area. One hundred twenty-five nurses and 28 home health aides participated. The methods of data collection included two written surveys conducted before and after intervention, chart audit tool and a case study analysis form. RESULTS: The precourse data revealed deficiencies in home-care agency staff knowledge in and clinical aspects of end-of-life care. Regarding agency nurses' knowledge on end of-life issues, the overall percentage scores from preeducation to posteducation rose from 84.6% to 89.0% (P =.0001). Participants rated the education program as extremely valuable for the home care agency staff. CLINICAL IMPLICATIONS: As home care will likely continue to be a primary setting for end-of-life healthcare, improvement of palliative care education in home health agencies is needed. The HOPE project provided insight and experience in the education of nonhospice home care staff in end-of-life care. Participant responses and evaluations indicated that end-of-life care education can improve the quality of care provided by home health agencies. Finally, this education program may result in a heightened awareness of hospice care and, although not objectively quantified, perhaps an increase in referrals to hospice. PMID- 11866707 TI - Healthcare professionals' awareness of cancer support services. AB - PURPOSE: This study was conducted to provide baseline information about the awareness, attitude, and willingness of oncology healthcare providers (HCPs) to recommend quality-of-life cancer education and support services to their patients. DESCRIPTION OF STUDY: A brief survey questionnaire was mailed to a random sample of 2000 physicians, nurses, and social workers who were active members of their respective oncology associations. RESULTS: A total of 1241 questionnaires were returned (62% reply rate), resulting in a final sample of 1180 HCPs (61% response rate). Among HCPs who referred patients to cancer organizations, the American Cancer Society (ACS; 83%), the National Cancer Institute (NCI; 55%), and the Leukemia and Lymphoma Society ([formerly Leukemia Society of America] 42%) ranked at the top. More than 70% of HCPs indicated that they had heard about specific ACS programs and services or the NCI information hotline. Less than 60% of HCPs recommended or thought these same services were helpful to patients. Awareness, referrals, and attitudes toward cancer support services among HCPs were significantly related (P <.001). Qualitative comments qualified quantitative results. CLINICAL IMPLICATIONS: Although a surprising number of HCPs were aware of support services, fewer recommended or reported satisfaction with these services. HCPs' referrals and attitudes toward cancer psychosocial support services may be improved by assembling a concise directory of locally available resources, directing this information to HCPs who refer directly to cancer organizations, improving collaboration between agencies, increasing staff support, and targeting particular services in specific areas of the country for program monitoring and development. PMID- 11866706 TI - Clinical trial enrollment of rural patients with cancer. AB - PURPOSE: The goal of this study was to examine the effect of a rural community clinical oncology program-based cancer-care intervention program that was launched to increase the number of rural patients with cancer enrolled in clinical trials. DESCRIPTION OF STUDY: Five rural counties in eastern North Carolina served as intervention communities, and five rural counties in South Carolina served as the comparison region. The intervention counties used a rapid tumor-reporting system, a nurse facilitator who identified and prompted oncologists to enter patients into clinical trials, a quarterly newsletter to primary-care physicians about cancer treatment and clinical trials, and a health educator who focused on community-wide education regarding cancer prevention, treatment, and clinical trial information. Outcomes included changes in knowledge and attitudes about clinical trials among the primary-care providers who were surveyed and enrollment in clinical treatment trials for breast and colorectal cancer, as analyzed by comparing practice pattern data from before and after the intervention. RESULTS: The results indicate that the intervention was not effective. The proportion of primary-care physicians who were aware of clinical trials for their patients with cancer rose slightly in comparison counties (26% to 34%) but remained constant (41% to 43%) in intervention counties. Perceived patient and actual physician barriers toward clinical trial participation were reported by the physicians. A minority of potentially eligible patients with breast or colon cancer in both North Carolina and South Carolina were enrolled in clinical trials. CLINICAL IMPLICATIONS: These data suggest that different types of interventions may be needed to improve accrual to cancer treatment trials in rural communities. In addition, the role that primary-care providers play in encouraging patients with cancer to participate in clinical treatment trials needs further exploration. PMID- 11866708 TI - Primary factor in declining breast cancer mortality rates: early detection or adjuvant therapy? PMID- 11866710 TI - Financial assistance from national organizations for cancer survivors. PMID- 11866712 TI - C-3prime prime or minute Quaternary Ammonium Cephems, a New Wave of Cephalosporins. PMID- 11866713 TI - On the Antimicrobial Activity of NO. PMID- 11866711 TI - Epidermal growth factor receptor inhibitors: a novel anticancer therapy. PMID- 11866714 TI - Retroviral Transmission and Breast-feeding. AB - Transmission of animal retroviruses has been demonstrated both for oncogenic retroviruses and animal lentiviruses. In humans, breast-feeding is the major route for mother-to-child transmission of Human T-cell Leukemia/Lymphoma Virus type I (HTLV-I). HTLV-I transmission by breast milk is associated with ingestion of infected cells and can be prevented by formula-feeding. Breast-feeding transmission of the Human Immunodeficiency Virus type 1 (HIV-1) has only been recently recognized as responsible for one to two thirds of mother-to-child transmission in breast-fed populations. A primary HIV-1 infection acquired in mothers after the baby has begun breast-feeding is associated with a particularly high risk of transmission. Breast milk transmission appears to result from the coexistence of HIV-1 and an inadequate humoral response in milk. Due to the dramatic impact of formula-feeding on child morbidity and mortality, it is suggested that present recommendation continue to promote breast-feeding in women living in settings where infectious diseases and malnutrition are the primary causes of infant deaths, as in many developing countries. On the other hand, in settings where infectious diseases and malnutrition are not the primary causes of infant deaths, as in most of the developed world, mothers with a proven HIV-1 infection should be advised not to breast-feed their babies. PMID- 11866715 TI - The Comparative In Vitro Activity of FK-037 (Cefoselis), a New Broad-Spectrum Cephalosporin. AB - FK-037, a new parenteral cephalosporin, was active against clinical isolates of both gram-positive and gram-negative aerobic bacteria. The activity of FK---037 was similar to that of cefpirome and cefepime and generally superior to that of ceftazidime. Methicillin-resistant staphylococci were less susceptible than methicillin-sensitive isolates but most remained within the sensitive range. All streptococci and Enterobacteriaceae were sensitive as were the majority of isolates of Acinetobacter spp. and Pseudomonas aeruginosa. There was resistance amongst other non-fermentative gram-negative bacilli but this was species specific and most pseudomonads were sensitive, with resistance seen in Stenotrophomonas maltophilia, Alcaligenes spp. and Flavobacterium spp. PMID- 11866716 TI - Rhodococcus equi Virulence-Associated Antigens and Specific Antibody Response in AIDS Patients Infected with R. equi. AB - OBJECTIVES: To analyze the expression of the 15- to 17-kDa plasmid-encoded antigens from Rhodococcus equi isolates of 7 AIDS patients and determine the immunologic response to these proteins in the patients' sera. METHODS: The expression of the virulence proteins in R. equi isolates and the specific antibody response were investigated by immunoblotting. Plasmid DNA was analyzed by agarose gel electrophoresis. RESULTS: The only patient infected with a strain carrying the virulence 85-kb plasmid and expressing the 15- to 17-kDa antigens developed a fatal pneumonia and did not produce specific antibodies to the virulence proteins. Of the 6 patients infected with R. equi strains lacking both proteins and plasmid, only 1 subject had a pulmonary disease with poor clinical outcome and exhibited a negligible humoral immune response to R. equi antigenic components, whereas the other patients who produced a remarkable antibody response developed either an asymptomatic infection (1 case) or pneumonia (4 cases) which completely cleared up. CONCLUSIONS: Our findings suggest that R. equi disease can be induced without the expression of the 15- to 17-kDa virulence associated plasmid-encoded antigens in HIV-infected patients with very low CD4+ cell counts. Nevertheless, both the synthesis of the virulence proteins and a defective humoral immune response to R. equi may contribute to the severity of rhodococcal disease. PMID- 11866717 TI - Antifungal Prophylaxis in Severely Neutropenic Patients: How Much Fluconazole is Necessary? AB - OBJECTIVES: To evaluate the efficacy of low dose fluconazole treatment for the prevention of yeast colonization and infection in severely neutropenic patients. METHODS: An open randomized trial, comparing fluconazole (100 mg per day) with nystatin (800,000 IU per day), in a University Hospital setting. RESULTS: Antifungal prophylaxis was given during the period of neutropenia, defined as less than 500 polymorphonuclear cells (PMN)/mm3). Thirty-six patients were randomly assigned to fluconazole and 33 to nystatin treatment groups. New oropharyngeal colonizations were significantly reduced by fluconazole (P=0.005), and oropharyngeal infections occurred less frequently in the fluconazole group (3% versus 16%, P=0.07). Stool colonization was identical between both groups. Systemic fungal infections were rare; one fluconazole patient had pulmonary aspergillosis and one nystatin patient developped Candida pseudotropicalis fungemia. Empiric amphotericin B was given with the same frequency in both groups. No side effects were associated with fluconazole. However, the administration of nystatin became impossible for three patients because of vomiting and lack of compliance. CONCLUSIONS: Fluconazole (100 mg per day) is more effective than nystatin for the prevention of oropharyngeal yeast colonization. Comparison with results in the literature suggests that a 100-mg dose of fluconazole has similar effects to 200 or 400 mg per day. PMID- 11866718 TI - Inhibition of Mononuclear Cell Procoagulant Activity by Lipophosphoglycan of Leishmania donovani. AB - BACKGROUND: Since fibrin formation is an expression of the response of the host to parasite spread, the lipophosphoglycan (LPG) of Leishmania donovani and its carbohydrate fragment (PG) were examined for their capacity to inhibit procoagulant activity (PCA) production by human mononuclear cells stimulated with Escherichia coli endotoxin in vitro. METHODS: the putative inhibitory effect of LPG and its PG fragment was evaluated on the basis of their in vitro capacity to prolong significantly the time required for coagulation induced by endotoxin stimulated mononuclear cells. RESULTS: LPG exhibited the most inhibitory activity, whereas the carbohydrate domain was not effective. These results are in agreement with the notion that LPG (but not PG) has an inhibitory effect on protein kinase C activity which plays a key role in the production of PCA by human monocytes. CONCLUSIONS: From a pathophysiological point of view, these data suggest the possibility that Leishmania avoids fibrin entrapment in the host through this inhibitory mechanism. PMID- 11866719 TI - Antimicrobial Activity of SCH 27899, Oligosaccharide Member of the Everninomycin Class with a Wide Gram-Positive Spectrum. AB - The in vitro antimicrobial activity of SCH 27899 (everninomycin), a novel oligosaccharide compound of the everninomycin class, was compared with vancomycin, chloramphenicol, clinafloxacin, teicoplanin and doxycycline against 428 clinical strains of bacteria. Everninomycin base exhibited the greatest antimicrobial activity compared to other formulations against all strains tested (MIC90: 0.25 microg/ml) followed by clinafloxacin and teicoplanin (MIC90: 0.5 microg/ml), vancomycin (MIC90: 2 microg/ml), and doxycycline (MIC90: 16 microg/ml). Everninomycin demonstrated the best activity against Streptococcus spp. (serogroups A, B, C, F, G) and Streptococcus pneumoniae, and lower activity against coagulase-negative staphylococci (MIC90: 0.5 microg/ml). All enterococci had an everninomycin MIC of 0.5 microg/ml or less. Everninomycin had no measurable antimicrobial activity against gram-negative aerobic organisms except Flavobacterium meningosepticum (MIC50: 2 microg/ml). Some everninomycin activity was observed against Clostridium spp., Peptostreptococcus spp., and the Prevotella bivius-disiens group. Everninomycin showed excellent activity (MIC90: 0.25 microg/ml) against the fluoroquinolone-resistant strains and all gram positive strains resistant to vancomycin (MICs less-than-or-equal 4 microg/ml). The MBC/MIC ratios and killing curve data suggest that everninomycin is not uniformly or rapidly bactericidal. These in vitro data indicate that everninomycin could be useful against emerging gram-positive strains resistant to other contemporary antimicrobials. PMID- 11866720 TI - Endocarditis Caused by Multiply Resistant Bacteroides fragilis: Case Report and Review. AB - A 78-year-old woman developed fatal endocarditis of her prosthetic aortic valve, caused by Bacteroides fragilis fragilis, and associated with ovarian carcinoma. The strain showed multiple antibiotic resistance, including resistance to beta lactam agents and combinations with beta-lactamase inhibitors. Seventeen previously described cases of endocarditis caused by Bacteroides spp. have been found in the literature. The mean age of the 18 patients was 50.3 years, the gastro-intestinal tract was the most common site of associated disease, embolism occured in ten cases and eight patients died. Previous isolates showed the antibiotic susceptibility customarily associated with the B. fragilis group. PMID- 11866721 TI - Evaluation of CrystalTM Rapid Stool/Enteric ID System for Identification of Aerobic Gram-negative Bacilli. AB - OBJECTIVE: The evaluation of the new miniaturized CrystalTM Rapid Stool/Enteric System (Becton-Dickinson, USA) for identification of aerobic gram-negative bacilli. METHODS: a total of 154 clinical organisms (Enterobacteriaceae: 120 strains; oxidase-positive fermenters: 13 strains; non-fermenters: 21 strains) were tested. Results were compared with those obtained with the PASCOR system (Difco, USA) and divergent identifications were evaluated by standard biochemical tests. RESULTS: without additional testing, correct identification was obtained for 146 strains (Enterobacteriaceae: 95%; oxidase-positive fermenters 87%; non fermenters 100%). For adequate identification of Yersinia enterocolitica strains, however, panels had to be incubated for 5 instead of 3 hours. CONCLUSIONS: the CrystalTM Rapid Stool/Enteric system offers a safe, accurate and rapid method for the identification of frequent isolates of the family Enterobacteriaceae and bacterial stool pathogens. PMID- 11866722 TI - Evaluation of the BBLreg CrystalTM MRSA ID System for Rapid Detection of Methicillin Resistance in Staphylococcus aureus. AB - Twenty-four clinical isolates of Staphylococcus aureus collected from various geographic areas and four reference strains were studied by (i) agar diffusion with disks impregnated with 5 microg oxacillin and reading after incubation at 30C for 24 hours, (ii) Southern hybridization with a probe specific for the mecA gene, and (iii) the BBLreg CrystalTM MRSA ID system. There was perfect correlation between the three methods: the BBLreg CrystalTM MRSA ID system detected methicillin resistance in the fifteen strains hybridizing with the mecA probe and classified as resistant by the oxacillin disk diffusion test; the thirteen remaining strains were susceptible by agar diffusion and by the BBLreg test and did not hybridize with the mecA probe. The BBLreg CrystalTM MRSA ID System, therefore, appears to be an accurate method for rapid detection of Staphylococcus aureus exhibiting homogeneous resistance to methicillin. PMID- 11866723 TI - Glycopeptide Resistance in Gram-positive Bacteria. PMID- 11866725 TI - Eugene Yourassowsky. PMID- 11866724 TI - In Vivo Selection of Streptococcus pneumoniae, Resistant to Quinolones, Including Sparfloxin. PMID- 11866726 TI - European Collaboration in Infectious Diseases Surveillance: Where to Go? PMID- 11866727 TI - Antibiotic Susceptibility Testing and Breakpoint Determinations. PMID- 11866728 TI - First Meeting of the Presidents of the European Medical Societies. PMID- 11866731 TI - LETTER FROM THE EDITOR-IN-CHIEF. PMID- 11866730 TI - LETTER FROM PRESIDENT OF THE SOCIETY. PMID- 11866732 TI - Animal models and Lyme disease. PMID- 11866733 TI - Skin and post-surgical wound infections due to Staphylococcus lugdunensis. PMID- 11866734 TI - Human microsporidiosis: Clinical, diagnostic and therapeutic aspects of an increasing infection. AB - Human microsporidiosis is a parasitic infection due to species of four different genera: Encephalitozoon; Enterocytozoon; Nosema; and Pleistophora. Although well known as a cause of disease in animals, microsporidiosis was only occasionally reported in humans. Recently, in human immunodeficiency virus (HIV)-infected patients, microsporidia belonging to Encephalitozoon and Enterocytozoon species have proved to be important opportunistic pathogens. Enterocytozoon bieneusi is associated with chronic intermittent diarrhea, cholangiopathy and sinusitis whereas Encephalitozoon intestinalis, Encephalitozoon hellem and Encephalitozoon cuniculi, the three Encephalitozoon species found in humans, are associated with diarrhea, rhinosinusitis, keratoconjunctivitis, nephritis and hepatitis. Diagnosis of microsporidial infections in humans was until recently an invasive, laborious procedure including electron microscopy of small intestine biopsies. However, new simple staining methods using Uvitex 2B or modified trichrome stain for feces and other body fluids have facilitated clinical diagnosis as well as drug evaluation and epidemiological studies. The application of monoclonal antibodies and molecular techniques such as the polymerase chain reaction have further improved microsporidial diagnosis. Treatment of Entero. bieneusi has, until now, been unsuccessful whereas albendazole has proved to be an effective treatment for Encephalitozoon species infection. Identification of effective treatment for Entero. bieneusi infections and further study of the pathogenicity of these microsporidial infections in immunocompetent hosts are important future challenges. PMID- 11866735 TI - Comparative in vitro activity of apalcillin alone and combined with Ro 48-1220, a novel penam beta-lactamase inhibitor. AB - OBJECTIVE: The in vitro activity of apalcillin plus Ro 48-1220, a novel penam sulfone beta-lactamase inhibitor, was compared with apalcillin alone, piperacillin/tazobactam, ticarcillin/clavulanic acid, amoxicillin/clavulanic acid, imipenem, ceftazidime and cefepime. METHODS: Agar dilution and broth microdilution testing of 854 bacterial strains, subcultured from frozen stocks incubated for 24 h in 5% carbon dioxide, was carried out to determine the minimum bactericidal (MBC) and minimum inhibitory concentrations (MIC) of each of the study drugs in accordance with the NCCLS M26-T method. RESULTS: Apalcillin/Ro 48 1220 was active against all gram-negative aerobic and anaerobic isolates except Klebsiella oxytoca (MIC90 32 microg/mL). Among the Enterobacteriaceae, synergy for apalcillin/Ro 48-1220 (4 microg/mL fixed concentration) vs apalcillin alone was demonstrated for nearly all species when comparing MIC90 results. Apalcillin/Ro 48-1220 was highly potent against beta-lactamase-producing Moraxella catarrhalis, Haemophilus influenzae and Neisseria gonorrhoeae (MICs less-than-or-equal 1 microg/mL). However, much of this activity was due to the direct antimicrobial action of Ro 48-1220 alone (MICs less-than-or-equal 4 microg/mL). All Pseudomonas aeruginosa, Stenotrophomonas (Xanthomonas) maltophilia and Acinetobacter species were inhibited by apalcillin/Ro 48-1220 (MIC90 0.25 to 4 microg/mL). For the aerobic gram-positive organisms, none of the drugs tested were consistently effective against oxacillin-resistant staphylococci, Corynebacterium jeikeium and Enterococcus species other than E. faecalis. Apalcillin/Ro 48-1220 was as effective as piperacillin/tazobactam against Escherichia coli producing extended-spectrum TEM enzymes, but less active against isolates producing SHV-type beta-lactamases. When tested against 204 ceftazidime-, gentamicin- or fluoroquinolone-resistant organisms, 78%, 91% and 66% of strains, respectively, were susceptible to apalcillin/Ro 48-1220 (less than-or-equal 16 microg/mL). CONCLUSIONS: Apalcillin/Ro 48-1220 is bactericidal with a modest inoculum effect; its wide spectrum of activity favors continued studies of spectrum, pharmacokinetics and in vivo efficacy. PMID- 11866736 TI - Granulocyte colony-stimulating factor (G-CSF) and interleukin (IL)-8 in sera from patients with Staphylococcus aureus septicemia. AB - OBJECTIVE: To determine the concentrations of granulocyte colony-stimulating factor (G-CSF) and interleukin (IL)-8 in sera from patients with Staphylococcus aureus septicemia and to correlate the results to peripheral neutrophil counts and the clinical outcome. METHODS: Serum samples from 64 consecutive patients with S. aureus septicemia were sequentially collected in a prospective study. RESULTS: The mean plus minus standard deviation (SD) serum G-CSF value on admission was 348 plus minus 830 with a range of 8 to 5400 pg/mL. G-CSF concentrations were elevated (> 76 pg/mL) in 38/64 patients (59%) as were serum IL-8 concentrations (> 67 pg/mL) in 23/64 patients (36%) on admission. The mean plus minus SD IL-8 value was 266 plus minus 422 pg/mL with a range of 2 to 1366 pg/mL. A correlation was found between serum IL-8 and white blood cell count on admission (p=0.008). CONCLUSIONS: Patients with uncomplicated septicemia frequently have elevated G-CSF values (84%) in comparison to patients with complicated septicemia (49%; p=0.02), indicating a possible protective effect of G-CSF in septic complications. PMID- 11866737 TI - Patterns of resistance of Mycobacterium tuberculosis in Spanish patients infected with human immunodeficiency virus type 1 (HIV-1). PMID- 11866738 TI - Survival of Chlamydia pneumoniae following contact with various surfaces. AB - OBJECTIVE: In this study, the survival and recovery of Chlamydia pneumoniae (Cp) strains TW-183, AR-39, AR-388 and CWL-029 were measured after inoculation on glass, stainless steel, FormicaR laminate, paper, fabric and human skin. METHODS: Inoculum in throat washes from healthy volunteers was applied to each surface. Samples were taken immediately after inoculum application and at specified intervals thereafter to determine infectivity. RESULTS: Infectious Cp was recovered from glass for up to 4 h, from paper and fabric for up to 3 h, from FormicaR laminate for up to 2 h, from stainless steel for up to 60 min and from human skin for up to 30 min. Drying of the inoculated area had no significant effect on the recovery of infectious Cp. Further experiments demonstrated that infectious Cp could be transferred to hands by touching these contaminated surfaces and could be recovered from these hands for up to 3 min. Addition of albumin, surfactant or phosphatidylcholine had no significant effect on the survival of Cp. CONCLUSIONS: These results suggest that contact with contaminated surfaces may be a potential mode of transmission of Cp. PMID- 11866739 TI - Risk factors and a clinical index for diagnosis of Pseudomonas aeruginosa bacteremia. AB - OBJECTIVE: To define risk factors significantly and independently associated with Pseudomonas aeruginosa bacteremia and to combine them in a diagnostic index which will define groups of septic patients at low or high risk of bloodstream infection caused by P. aeruginosa. METHODS: Logistic regression analysis was used to identify risk factors associated with pseudomonal bacteremia among all patients with bacteremia at St Thomas' Hospital, London, from 1969 to 1989. The coefficients of the final logistic model were used to compile a linear model for the diagnosis of pseudomonal bacteremia. The index was validated in another set of patients with bacteremia detected at Beilinson Medical Center, Petah Tiqva, Israel, from 1988 to 1991. RESULTS: Seven factors were significantly and independently predictive of pseudomonal bacteremia: 1) neutropenia; 2) previous or current treatment with antibiotics; 3) cytotoxic or corticosteroid treatment; 4) hospital acquisition of infection; 5) detection in the intensive care unit; 6) male gender; and 7) focus of infection. High-risk foci were the urinary tract with catheter or post-instrumentation, or unknown source. Low-risk foci were bone, joint, meninges, female genital tract and upper respiratory tract. The index score divided patients into three groups with increasing likelihood of P. aeruginosa bacteremia: 1%, 7% and 19%, respectively (p=0.0001). In the validation set, the percentage of P. aeruginosa bacteremia in the three respective groups defined by the index were 5%, 18% and 22% (p=0.0001). CONCLUSIONS: The use of simple clinical and laboratory data known within hours of detection of an infectious episode can define patients at high and low risk for P. aeruginosa bacteremia. PMID- 11866740 TI - Fungemia: An increasing problem in a Danish university hospital 1989 to 1994. AB - OBJECTIVE: To present data on episodes of fungemia in a Danish tertiary-care university hospital admitting all types of patients and to compare the data with previous findings from the same hospital. METHODS: Retrospective identification of episodes of fungemia from 1989 to 1994 and collection of data from computerized files at the Clinical Microbiology Department at Rigshospitalet and the Mycology Reference Laboratory at Statens Seruminstitut. RESULTS: The incidence of fungemia increased gradually from 19 episodes in 1989 to 57 episodes in 1994. An earlier report from the same hospital showed 20 to 25 episodes of fungemia per year between 1984 and 1988. Candida albicans was the dominating species during both periods, accounting for 73% of isolates during 1984 to 1988 and 67% during 1989 to 1994. However, in the hematology department where fluconazole has been used extensively, C. albicans constituted 47% of isolates with Candida krusei and Candida glabrata comprising 25%. CONCLUSIONS: The incidence of fungemia in our tertiary-care hospital has increased threefold from 1989 to 1994. Candida albicans was the dominating cause of fungemia but, in the hematology department, this yeast accounted for less than half of the isolates during the same time period. PMID- 11866741 TI - Comparison of macrorestriction analysis of genomic DNA by pulsed-field gel electrophoresis and ribotyping with conventional methods for differentiation of Escherichia coli 0124 isolates. AB - OBJECTIVE: To evaluate the usefulness of different phenotypic and genotypic markers for epidemiological typing of enteroinvasive Escherichia coli 0124 (EIEC 0124). METHODS: Seven sporadic EIEC 0124 isolates and 22 isolates from two different outbreaks were characterized. Chromosomal deoxyribonucleic acid (DNA) macrorestriction analysis with XbaI resolved by pulsed-field gel electrophoresis (PFGE) and ribotyping with each of the three restriction endonucleases BglII EcoRI, and ClaI were compared with biotyping, antimicrobial susceptibility patterns and plasmid profiles. RESULTS: Biotypes and antimicrobial resistance profiles of the outbreak-associated strains showed considerable variation, thereby limiting the usefulness of such phenotypic markers. Only 57% of the sporadic isolates harbored plasmids. Three different ribotypes based on 5 to 7 bands were recorded among sporadic isolates whereas all outbreak-associated strains showed the same ribotype. BglII appeared to give the best discrimination whereas EcoRI and ClaI provided no additional information. Sporadic EIEC 0124 isolates showed a marked diversity of macrorestriction patterns (similarity coefficient 58 to 93%) and five different patterns were detected. In contrast, the outbreak isolates were closely related (similarity coefficient 90 to 100%). Genomic DNA macrorestriction analysis correlated well with ribotyping, but PFGE was more discriminating. CONCLUSIONS: PFGE is a useful method for epidemiological comparison and differentiation of EIEC 0124 isolates. PMID- 11866742 TI - Evaluation of the ATB 32 C system for identification of clinical yeast isolates. AB - OBJECTIVE: To compare the ATB 32 C system for routine identification of clinical yeast isolates in a clinical microbiology laboratory with identification carried out by conventional methods in a mycology reference laboratory. METHODS: A total of 113 strains initially isolated at our hospital and identified in the reference laboratory were returned in duplicate, under separate code numbers, to the microbiology laboratory where the ATB 32 C system was used for identification by: 1) visual assessment of turbidity at 72 h with use of identification table; 2) visual assessment at 72 h with use of ATB 32 C analytical profile index; and 3) automatic readings with the ATB reader at 48 h and 72 h with results of growth assessments transmitted to a computer and interpreted by the ATB 32 C software. RESULTS: Visual assessment plus identification table and visual assessment plus profile index provided correct identification in 98% and 91% of strains, respectively. Visual assessment was, however, sometimes difficult and required more experience than is usually available in a routine clinical microbiology laboratory. Automatic readings with computer identification plus supplementary tests correctly identified 87% and 86% after 48 h and 72 h, respectively. CONCLUSIONS: The ATB 32 C system with automatic readings and computer identification is a satisfactory system for identification of clinical yeast isolates in a routine clinical microbiology laboratory. PMID- 11866743 TI - Diphtheria in the 1990s: Return of an old adversary. PMID- 11866744 TI - Epidemic vancomycin-resistant Enterococcus faecium in the UK. PMID- 11866746 TI - Ehrlichia species as possible causative agents of blood culture-negative endocarditis. PMID- 11866745 TI - Listeriosis associated with pregnancy and human immunodeficiency virus (HIV) infection. PMID- 11866747 TI - Penicillin resistance in Streptococcus pneumoniae in Istanbul, Turkey. PMID- 11866748 TI - Endocarditis with negative blood cultures. PMID- 11866749 TI - Microsporidia and other opportunistic protozoa in patients with acquired immunodeficiency syndrome (AIDS). PMID- 11866750 TI - Value of the hospital epidemiologist. PMID- 11866751 TI - Secular trends in mortality associated with bloodstream infections in 4268 patients hospitalized in a cancer referral center between 1975 and 1989. AB - OBJECTIVE: To study the trends in mortality over 15 years in hospitalized cancer patients with bloodstream infection. METHODS: The yearly incidence rates and risk of death, by type of microorganism, were calculated for 4268 cancer patients hospitalized between 1975 and 1989 in a French cancer referral center. The relative risk of death (RR) associated with each type of microorganism was estimated using the proportional hazards model, taking into account age, hospital ward, underlying disease, geographical origin and year of the first positive blood culture. RESULTS: The incidence of these infections was five-fold higher in 1989 than in 1975. The largest increases were for coagulase-negative staphylococci (CNS), yeasts and Staphylococcus aureus. For the 3756 patients who had a single-microorganism bloodstream infection, the risk of death compared with that of patients with CNS infection was significantly increased in those with Pseudomonadaceae (RR=5.0), yeasts (RR=3.4), Enterobacteriaceae (RR=3.2), S. aureus (RR=2.8) and streptococci (RR=2.1). The risk of death was not significantly different between patients with a single or several positive blood cultures nor between those with nosocomial or non-nosocomial infections. When the study period was divided in two time periods (1975 to 1982 vs 1983 to 1989), a significant variation (p=0.001) in risk of death associated with the different microorganisms was observed. Most risks were lower from 1983 to 1986 than before 1982. This decrease reached 60% for both S. aureus and Pseudomonadaceae. CONCLUSIONS: These data support of continuing use of aggressive empirical antimicrobial therapy for cancer patients with fever. PMID- 11866752 TI - Antibiotic susceptibilities of Haemophilus influenzae in central Scotland. AB - OBJECTIVE: To ascertain the incidence of antibiotic resistance in Haemophilus influenzae in central Scotland and the beta-lactamases produced by these isolates. METHODS: A total of 213 H. influenzae isolates from four medical centers in Scotland [Aberdeen (n=58), Edinburgh (n=55), Glasgow (n=64) and Dundee (n=36)] were tested for susceptibility to a range of antimicrobials including beta-lactams, beta-lactam/beta-lactamase-inhibitor combinations, and a representative 4-quinolone, antifolate and macrolide. Susceptibility testing of the beta-lactam/beta-lactamase-inhibitor combination amoxicillin plus clavulanic acid was conducted at both 2:1 and 4:1 ratios and with clavulanic acid fixed at a concentration of 2 mg/L. Each strain was further investigated for the presence of beta-lactamase activity. RESULTS: Although the incidence of resistance to amoxicillin was 15%, in the presence of clavulanic acid, this resistance was reduced to 4.2%, 5.6% and 4.2% with the 2:1 ratio, 4:1 ratio and 2 mg/L fixed concentration, respectively. Sixteen percent of the isolates demonstrated immediate beta-lactamase production. Isoelectric focusing showed that 77.4%, 16.1% and 6.5% of the beta-lactamase-positive strains were found to contain TEM 1, VAT-1 and both TEM-1 and VAT-1 beta-lactamases, respectively. A further 29% of the strains were recognized as being beta-lactamase-positive after prolonged incubation with nitrocephin. CONCLUSIONS: This study suggests that current testing for beta-lactamases may underestimate the prevalence of beta-lactamase production in H. influenzae. PMID- 11866753 TI - Screening for Escherichia coli O157:H7 in Illinois. AB - OBJECTIVE: To determine the incidence of infection with Escherichia coli O157:H7 in a tertiary referral center in Chicago, where a similar study had been performed in 1984, to evaluate cases of disease reported to the Illinois Department of Public Health (IDPH) in 1993, and to determine laboratory practices used to detect this infection throughout the state. METHODS: During a 6-month period in 1993, all stool specimens at Rush-Presbyterian-St Luke's Medical Center (RPSLMC) were tested for E. coli O157:H7. Reports of diagnosed E. coli O157:H7 cases investigated by IDPH were also reviewed. A survey of 73 hospitals in the Chicago area was performed to determine routine culturing practices, specifically, the selection of stool specimens for evaluation for this pathogen. RESULTS: In the RPSLMC survey, two cases were identified among 1985 samples (incidence 0.1%), similar to the 0.08% incidence detected in a similar study conducted at the same institution in 1984. Through passive surveillance, the IDPH received 44 reports of E. coli O157:H7 in 1993. The hospital survey revealed that, in the seven labs testing all stool specimens for E. coli O157:H7, an incidence of 16/8137 specimens (0.2%) was determined. CONCLUSIONS: These data suggest that sporadic E. coli O157:H7 remains uncommon in Illinois and that the incidence may not have changed over a 9-year period. The low yield and substantial cost of culturing all stools suggest that only specimens from patients with bloody diarrhea should be evaluated routinely in areas of low endemicity. PMID- 11866754 TI - Enhancement of growth of group A beta-hemolytic streptococci in mixed infections with aerobic and anaerobic bacteria. AB - OBJECTIVE: To evaluate the potential for mutual enhancement of growth of group A beta-hemolytic streptococci (GABHS), and 11 commonly associated aerobic and anaerobic microorganisms frequently isolated in tonsillar infections. METHODS: Enhancement was assessed by measuring the relative increase in colony-forming units (CFU) of GABHS and each of the 11 microorganisms inducing a subcutaneous abscess in mice. RESULTS: Of the 11 combinations of GABHS and aerobe or anaerobe, GABHS was enhanced in 10 cases and the other microorganism in 5, namely, Staphylococcus aureus, Haemophilus influenzae type b, Klebsiella pneumoniae, Prevotella melaninogenica and Bacteroides fragilis. CONCLUSIONS: These findings confirm the mutual symbiotic enhancement of growth of GABHS in the presence of other aerobic and anaerobic bacteria. PMID- 11866755 TI - In-vitro interaction of azithromycin and fluoroquinolones against gram-positive and gram-negative bacteria. AB - OBJECTIVE: To determine the combined in-vitro effects of azithromycin plus the fluoroquinolone ofloxacin or lomefloxacin against gram-positive and gram-negative bacteria. METHODS: Fractional inhibitory (FIC) and fractional bactericidal concentration indices of azithromycin and the fluoroquinolone were determined using a microtiter-checkerboard method. Clinical isolates of Staphylococcus aureus, Streptococcus pneumoniae, Neisseria gonorrhoeae, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Pseudomonas cepacia, Haemophilus influenzae, Xanthomonas maltophilia and Acinetobacter calcoaceticus were studied. Fourteen strains of S. aureus were also studied in time-kill curves with azithromycin (4 mg/L), lomefloxacin (6 mg/L) and the two in combination. RESULTS: No synergism or antagonism was found in inhibitory assays. However, bactericidal assays revealed antagonism with some strains of S. aureus, S. pneumoniae, X. maltophilia, A. calcoaceticus, P. aeruginosa, P. cepacia, K. pneumoniae and E. coli. Kill-curve results with 14 strains of S. aureus showed no antagonism with four strains of methicillin-resistant S. aureus (MRSA), and antagonism with one strain of MRSA and seven methicillin-susceptible S. aureus (MSSA). CONCLUSIONS: In-vitro exposure to combinations of azithromycin and a fluoroquinolone does not produce a synergistic effect. Antagonism was found in bactericidal assays against some gram-negative bacteria and MSSA; caution is therefore recommended in the use of macrolides and quinolones against these organisms. PMID- 11866756 TI - Reevaluation of contemporary laboratory methods for detection of antimicrobial resistance among enterococci. AB - OBJECTIVE: To evaluate broth microdilution, disk diffusion, Etest and Vitek Systems for susceptibility testing of enterococci. METHODS: Susceptibility testing of a panel of 149 enterococci (99 vancomycin-resistant) strains, using the study methods, was performed and the results compared. RESULTS: For vancomycin susceptibility testing, categorical agreement of disk diffusion, Etest and Vitek with the reference broth microdilution test was > 95%. For aminoglycoside and ampicillin testing, categorical agreement between Etest and Vitek was 98 to 100%. CONCLUSIONS: Disk diffusion, Etest and Vitek have acceptable performance for detection of vancomycin resistance of Van A and Van B phenotypes among enterococci. PMID- 11866757 TI - Comparison of polymerase chain reaction and pp65 antigen test for early detection of human cytomegalovirus in blood leukocytes of cardiac transplant recipients. AB - OBJECTIVE: To establish whether polymerase chain reaction (PCR) for cytomegalovirus deoxyribonucleic acid (DNA) can provide clinical information for the management of the infection. METHODS: Leukocytes in 30 heart transplant recipients were monitored by pp65 antigen testing and PCR for 82 to 365 days after transplantation. RESULTS: Of the 30 patients, 26 developed cytomegalovirus infection, nine of whom were symptomatic. Altogether, 300 leukocyte samples were examined. The concordance between PCR and pp65 antigen test was 82.6%. In symptomatic patients after surgery, PCR detected cytomegalovirus infection after 38 plus minus 16 days and the pp65 antigen test, after 48 plus minus 15 days. Symptomatic infection correlated with a higher number of pp65-positive leukocytes than did asymptomatic infection: 310 plus minus 356 vs 24 plus minus 35 (p < 0.005)/200,000 examined, respectively. Clearance of virus was observed by PCR after 125 plus minus 73 days (range 29 to 225) in symptomatic, and after 82 plus minus 70 days (range 16 to 301) in asymptomatic, cases of infection. CONCLUSIONS: The positive predictive value of PCR for symptomatic infection was 34.6%. Our findings correlate with previous reports and show that the qualitative detection of cytomegalovirus DNA is not associated with overt disease whereas quantitation of pp65-positive leukocytes closely correlate with symptom onset. Insofar as the results are not quantitative, PCR is not a marker of clinically apparent infection. Careful monitoring of cytomegalovirus infection based on quantitative pp65 antigen assay can fulfill all clinical needs for early diagnosis and proper management of the infection PMID- 11866758 TI - Facilitating learning and change in physicians: Implications for a system of continuing medical education in Europe. PMID- 11866759 TI - Antibiotic susceptibility testing and breakpoint determination. PMID- 11866761 TI - Risk-factor analysis and diagnostic index for Pseudomonas aeruginosa bacteremia. PMID- 11866760 TI - Continuing medical education (CME) credits and multiple-choice questions. PMID- 11866762 TI - Malignant ventricular arrhythmia during treatment with terfenadine and itraconazole: A pharmacokinetic study. PMID- 11866763 TI - In-vitro release of vancomycin and netilmicin from bone cement. PMID- 11866764 TI - Thrice-weekly maintenance therapy for cytomegalovirus retinitis in patients with acquired immunodeficiency syndrome (AIDS). PMID- 11866765 TI - Infection caused by Ochrobactrum anthropi. PMID- 11866766 TI - Review of meeting: The envelope in bacterial physiology and antibiotic action. PMID- 11866767 TI - Review of publication: Coping with Change. PMID- 11866768 TI - Get ready for the next influenza pandemic ! PMID- 11866769 TI - Cycling of antibiotics: an approach to circumvent resistance in specialized units of the hospital. PMID- 11866770 TI - Antimicrobial peptides: broad-spectrum antibiotics from nature. PMID- 11866771 TI - Aseptic meningitis after neurosurgery: a demonstration of bacterial involvement. AB - OBJECTIVE: To evaluate the presence of bacteria in samples from patients suffering from 'aseptic' meningitis following craniotomy. METHODS: Prospective study in which cerebrospinal fluid (CSF) from patients suffering from post craniotomy meningitis and negative control patients were submitted to conventional culture and to polymerase chain reaction (PCR) using bacterial 16S rRNA universal primers, followed in some cases by DNA sequencing of the PCR product and phylogenetic analysis. RESULTS: CSF from patients with either culture positive or culture-negative meningitis yielded positive amplifications, whereas no amplification was obtained with CSF from control patients. All positive signals were confirmed by Southern hybridization with a prokaryote 16S RNA specific probe. Six PCR products, of which three were collected from later cases of culture-negative meningitis, were cloned and sequenced. Sequence analysis suggested affinities with Pseudomonas in three cases, with Escherichia in two cases and with Rhodococcus in one case. CONCLUSIONS: Many cases of culture negative (aseptic) meningitis are probably bacterial meningitis and justify antibiotic treatment. The bacteria responsible for these cases of culture negative meningitis might have peculiar growth requirements in vitro. PMID- 11866772 TI - Comparative pharmacokinetics of macrolide antibiotics and concentrations achieved in polymorphonuclear leukocytes and saliva. AB - OBJECTIVES: The pharmacokinetics of macrolide antibiotics --- erythromycin (ER), clarithromycin (CL), roxithromycin (RO), azithromycin (AZ), dirithromycin (DI) and the concentrations achieved in polymorphonuclear neutrophils (PMNs) and saliva were investigated. METHODS: In a four-way crossover trial, 10 healthy volunteers received 1000 mg ER twice a day, 500 mg CL twice a day, 150 mg RO twice a day and 500 mg AZ every day over a period of 3 days. In a second trial, 10 healthy volunteers received 500 mg DI every day over a period of 5 days. Concentrations of these antibiotics were measured in serum, urine, saliva and PMNs by high-performance liquid chromatography (HPLC) on days 1 and 3 in the first trial and on days 1 and 5 in the second trial. RESULTS: We found considerable differences in the pharmacokinetics, not only in serum, but also in PMNs and saliva. All substances except RO exhibited higher concentrations in PMNs than in serum, indicating excellent intraphagocytic distribution. In contrast, concentrations in saliva were lower than those measured in serum, with the exception of AZ. ER is characterized by low serum concentrations and moderate concentrations in saliva and PMNs. CL reached considerable concentrations in serum, saliva and PMNs. RO achieved the highest serum levels, but concentrations in saliva and in PMNs were below the detection limit. In contrast, AZ and DI yielded the lowest serum concentrations and the highest saliva and PMN concentrations. CONCLUSIONS: Our data emphasize the importance of tissue distribution, in addition to serum kinetics, in evaluating the pharmacokinetic profiles of antibiotics. PMID- 11866773 TI - Effect of subinhibitory concentrations of antimicrobial agents on adherence to silicone and hydrophobicity of coagulase-negative staphylococci. AB - OBJECTIVES: To study the effect of 0.25 MIC of antimicrobial agents on adherence to silicone and hydrophobicity of a slime-producing Staphylococcus epidermidis (ATCC 35984) and non-slime-producing Staphylococcus hominis (ATCC 35982). METHODS: Adherence was assessed in vitro, using silicone rubber immersed in a suspension of bacteria, pretreated with 0.25 MIC of oxacillin, ceftriaxone, vancomycin or pefloxacin. After a 24-h period, adherent bacteria, detached by trypsin and sonication, were counted. Hydrophobicity was assessed by measuring the affinity of pretreated bacteria to p-xylene. RESULTS: For slime-producing S. epidermidis, adherence was significantly decreased by 81%, 91% and 77% with oxacillin, vancomycin and pefloxacin respectively. For non-slime-producing S. hominis, adherence was significantly decreased by 75% and 94% with oxacillin and ceftriaxone respectively. Hydrophobicity of both strains was significantly decreased with oxacillin only. CONCLUSIONS: Adherence of coagulase-negative staphylococci onto silicone can be modified by sub-MICs of some of the antimicrobial agents tested. This effect was different in the slime-producing and non-slime-producing strains, and was not correlated with the mechanism of the inhibitory effect of these antimicrobial agents, or the modification of hydrophobicity. This suggests that some surface components, not involved in hydrophobicity, could play a role in in vitro adherence to silicone. PMID- 11866774 TI - Asymptomatic oral yeast carriage in HIV-infected patients: frequency and fluconazole susceptibility profile. AB - OBJECTIVES: Fluconazole-resistant oropharyngeal candidiasis (OPC) is a rapidly growing problem in HIV-infected patients. To better understand the pathogenesis of fluconazole resistance in this setting, asymptomatic candidal carriage was determined by means of oral swabs regularly performed in all patients without clinical signs of OPC seen at our HIV outpatient clinic. Controls were 204 asymptomatic healthcare workers without previous exposure to fluconazole. METHODS: Swabs were plated on three solid media and put in a Sabouraud broth. Phenotypically different colonies were identified to the species level. Susceptibility to fluconazole was determined using a disk diffusion test with 50 microg fluconazole disks on yeast nitrogen agar, with a cut-off value of 25 mm. RESULTS: Swabs were performed in 538 consecutive HIV-positive patients, of whom 216 (40%) had had prior episode(s) of OPC and/or were previously exposed to fluconazole. Yeasts were grown in 418/538 HIV-positive patients (78%), compared to 57/204 controls (28%) (p < 0.05). In HIV-positive patients, yeasts were grown in 189/216 (88%) of those with past fluconazole exposure, and in 229/322 (71%) without exposure (p < 0.05). A total of 589 isolates were grown in the 538 HIV positive patients (451 C. albicans, 88 C. glabrata, 22 C. tropicalis, 11 C. krusei, and 17 isolates from 12 other species). Resistance to fluconazole was present in 121/589 (21%) Candida species isolates in HIV-positive patients and in 2/59 (3%) in controls. Among C. albicans isolates, there were 18 fluconazole resistant strains in HIV-positive patients (4%) and none in controls.CONCLUSIONS: Using sensitive culture methods, oral yeast colonization was detected significantly more frequently in HIV-infected patients (78%) than in a control group of HIV-negative persons (28%). In addition, yeast colonization was quantitatively more important in patients with lower CD4+ lymphocyte counts and for those who had been exposed to fluconazole for episode(s) of OPC. Fluconazole resistant C. albicans isolates were observed only in HIV-positive patients, and all patients (17/18) for whom this information could be ascertained had had prior exposure to fluconazole. PMID- 11866775 TI - Prevalence of Chlamydia trachomatis, Chlamydia psittaci and Chlamydia pneumoniae antibodies in blood donors and attendees of STD clinics. AB - OBJECTIVE: To estimate the prevalence of Chlamydia trachomatis, C. psittaci and C. pneumoniae antibodies in sera from altogether 931 blood donors, patients with symptoms of urethritis, assumed salpingitis and sexually acquired reactive arthritis (SARA), and women with fertility problems. METHODS: IgG antibodies to C. trachomatis, C. psittaci and C. pneumoniae were determined by microimmunofluorescence (MIF) tests. All patients were also tested for genital C. trachomatis infection using direct immunofluorescence (DIF) tests. RESULTS: The DIF-positive cases had a significantly (p < 0.0001) higher prevalence of C. trachomatis antibodies than the DIF negatives, i.e. 88.5% versus 14% in men with urethritis, 94.3% versus 36.4% in women with salpingitis, 66.7% versus 16.7% in SARA patients and 90.6% versus 20.8% in women with fertility problems. Antibody reactivity to all three chlamydial species was found significantly (p < 0.0001) more often in the patient groups and in those with a DIF-confirmed genital C. trachomatis infection than in blood donors. CONCLUSIONS: Presence of serum antibodies to C. trachomatis is tightly associated with the presence of chlamydiae in the genital tract, which also influences the cross-reactivities occurring in the MIF tests between chlamydial species. PMID- 11866777 TI - Infections in asplenic patients. PMID- 11866776 TI - Typing of Pseudomonas aeruginosa strains from patients with cystic fibrosis: phenotyping versus genotyping. AB - OBJECTIVE: To assess the discriminatory power of two genotypic and two phenotypic techniques by analysis of Pseudomonas aeruginosa sputum isolates obtained with long term intervals from 29 independent cystic fibrosis (CF) patients. METHODS: Fifty-eight strains of P. aeruginosa were subjected to serotyping and pyocin production was assessed. Arbitrarily primed polymerase chain reaction (AP PCR) and pulsed-field gel electrophoresis (PFGE) were applied in order to detect genetic polymorphisms. RESULTS: From the results of different typing techniques, it appeared that the numbers of separate types varied between 11 and 43, and the percentage of identical P. aeruginosa pairs from individual patients varied between 51% and 72%, depending on the test system used. AP PCR and PFGE displayed enhanced resolution when compared to serotyping and pyocin typing; both DNA typing techniques generated concordant results, although differences in resolution are apparent. This results in 15% discordance, which may be the result of differences in the definitions of (sub)clonal relatedness as applied for AP PCR and PFGE, respectively. CONCLUSIONS: Molecular typing techniques are superior to phenotyping where P. aeruginosa is concerned. AP PCR is a fast and useful technique for determining clonality among P. aeruginosa strains from chronically colonized CF patients. It is clear, however, that the interpretation of data and comparative analysis of PFGE and AP PCR results necessitates additional (international) standardization and the development of practical guidelines. PMID- 11866778 TI - In-vitro acquisition of iron from the iron---desferrioxamine complex by Aeromonas hydrophila. PMID- 11866779 TI - Bartonella (Rochalimaea) quintana endocarditis in an Algerian farmer. PMID- 11866780 TI - Neonatal bacteremia caused by Agrobacterium radiobacter: a case and a review of the pediatric literature. PMID- 11866781 TI - A case of severe adenovirus pneumonia in a neonate. PMID- 11866783 TI - Review of meeting: Antibiotic discovery: Exploiting new understanding of mechanism of action. PMID- 11866784 TI - Tuberculosis as an AIDS-defining disease in Europe. PMID- 11866785 TI - Definition of intracellular pathogens. AB - Most microorganisms are destroyed by the host tissues through processes which usually involve phagocytosis and lysosomal disruption. However, some organisms are capable of growing inside macrophages and avoiding destruction. This paper describes the normal phagocytic process and how it is avoided by certain viruses, bacteria and protozoa, and the complex cycle of intracellular replication by species of Chlamydia. PMID- 11866787 TI - DISCUSSION. PMID- 11866786 TI - Diagnostic methods for intracellular pathogens. AB - Diagnosis of infection with Chlamydia pneumoniae is difficult and the optimal diagnostic procedure has yet to be established. C. pneumoniae is more difficult to isolate in tissue culture than C. trachomatis. Attention must be paid to the site from which the specimen is taken, and all specimens should be processed within 24 h or stored at 4 C. Serologic diagnosis largely depends on microimmunofluorescence testing which, despite proposed criteria, still has a large subjective component. Serology has limitations in terms of both sensitivity and specificity and antibodies may be difficult to detect in individuals with positive C. pneumoniae cultures. Non-culture methods include enzyme immunoassays, fluorescent-antibody techniques and DNA probes. The most promising appears to be PCR. Co-infection of C. pneumoniae and other respiratory organisms seem to be common. PMID- 11866788 TI - Epidemiology of intracellular pathogens. AB - Epidemiologic studies of Chlamydia pneumoniae have been conducted throughout the world. A rate of antibody prevalence of approximately 50% has been found in adults, with small regional differences. We studied the antibody prevalence of C. pneumoniae in healthy persons and patients with acute respiratory infections in Japan. The microplate immunofluorescence antibody (MFA) technique was used for measuring antibodies, and the cases with IgG titers of greater-than-or-equal 1:64 were considered to have had past chlamydial exposure. Using this criterion, positive serum antibodies against C. pneumoniae were found in 67.4% of healthy subjects and 74.2% of adult patients with respiratory infections. Some seroepidemiologic studies in Western countries have shown that epidemics of C. pneumoniae respiratory infections occur in a 4--6-year cycle, like a Mycoplasma pneumoniae infection. However, during our 7-year survey from 1988 to 1994, we were not able to observe any epidemic episodes. The prevalence of the antibody was lower than 10% in children 5 years old or younger. It increased rapidly between ages 6 and 15, reaching a plateau of 60%. Results suggest that in Japan a high infection rate might occur in children at nursery schools, kindergartens and elementary schools. We encountered an outbreak of C. pneumoniae infection and we were able to trace the source of the infection to a family and the spread of the infection to the schools which the children attended. Transmission is of the non vector type. The incubation period is 3 to 4 weeks. Transmission occurs only after repeated and close contacts. Small outbreaks may occur in households and schools where persons have prolonged close contacts. Unlike acute viral infections, it may spread slowly. PMID- 11866789 TI - Clinical features of Chlamydia pneumoniae acute respiratory infection. AB - Chlamydia pneumoniae is a worldwide respiratory pathogen involved in 6--20% of community-acquired pneumonias and in about 5% of acute exacerbations of chronic bronchitis. Preliminary data also indicate a possible association between Chlamydia pneumoniae infection and asthma. Further studies are needed to elucidate whether Chlamydia pneumoniae is merely a precipitant of asthma symptoms or is actually one of the causes of asthma. PMID- 11866790 TI - Chlamydia pneumoniae and cardiovascular diseases. AB - Chlamydia infection of the cardiovascular system is associated with pericarditis, endocarditis and myocarditis. Chlamydia particles can also be observed in damaged heart valves. There is now good evidence that the lesions of arteriosclerosis and aortic aneurysm as well as valvular disease may be associated with C. pneumoniae infection. Patients with acute myocardial infarction show seroconversion against Chlamydia lipopolysaccharide. In a prospective study of 4000 healthy hypercholesterolemic men, signs suggestive of chronic C. pneumoniae infection increased the risk of a cardiac event three---fold. This risk factor is synergistic with the smoking risk. Immunohistochemistry also demonstrated Chlamydia lipopolysaccharide in samples of aortic aneurysm. Chlamydial inflammation may play a role in the oxidation of low density lipoprotein in atherosclerotic lesions. PMID- 11866791 TI - DISCUSSION. PMID- 11866793 TI - Future trends in Chlamydia pneumoniae research and the place of macrolides in treatment. PMID- 11866792 TI - Intracellular bioactivity of macrolides. AB - A brief overview is provided of the bioactivity of macrolides against a range of bacterial species. Topics considered include the cellular pharmacokinetics of uptake and efflux of these drugs and the importance of intra- or extracellular and cytoplasmic or granular location on their activity. Emphasis is placed on the importance of synergy between macrolides and host defenses, with drug accumulation producing modification of cellular function, such as enhancement of phagocytosis, and exocytosis of polymorphonuclear neutrophils. Such interaction may explain the activity of such agents against organisms which normally inhibit fusion of phagolysosomes. PMID- 11866794 TI - DISCUSSION. PMID- 11866795 TI - Chairman's Introduction. PMID- 11866796 TI - Community-acquired pneumonia: epidemiologic and clinical considerations. AB - An incidence of between 2 and 44 per 1000 population has been reported for community-acquired pneumonia. Epidemiologic studies describe a wide range of causative organisms, including Streptococcus pneumoniae, Haemophilus influenzae, Mycoplasma pneumoniae, Legionella spp., Moraxella catarrhalis, Chlamydia pneumoniae and viruses such as influenza A and B. However, the frequency with which they are reported varies widely. On analysis of these studies, the variation can be explained by a number of factors. The results depend on the definition of pneumonia and the criteria for assigning a causative role to any particular organism. Older studies have not included diagnostic methods for newly described organisms such as C. pneumoniae and Legionella spp. The improved diagnostic methods for these organisms and for Mycoplasma pneumoniae are reflected in more recent studies. Further variation depends on the population studied. As many patients with mild pneumonia are successfully treated in the community, those studies that are hospital-based include patients with more severe pneumonia often in the elderly or in patients with underlying diseases such as chronic obstructive pulmonary disease. The prior use of antibiotics not only contributes to the high percentage of cases for which no etiologic agent is found, but also ensures that treatment failures are selected for hospitalization. This further changes the result, depending on the antibiotic agents used most commonly in the community. The inclusion of nursing home patients or groups where alcoholism is more common will also favor particular organisms. Finally, the timing of the study may be such that an epidemic is included. This has relevance mostly for Mycoplasma pneumoniae, C. pneumoniae, Legionella spp. and influenza. In the assessment of the patient with community-acquired pneumonia, any one of the above organisms can be considered to be responsible. As initial treatment is empirical, other information can be used to ensure that an antibiotic with an adequate spectrum is chosen. Factors of importance are age, underlying illness, severity of disease and any locally recognized epidemics or endemic organisms. Differences in clinical presentation are not sufficiently distinct to allow for accurate prediction of the causative agent. Similarly, chest radiograph changes are not sufficiently specific to discriminate reliably between diverse organisms such as S. pneumoniae, Mycoplasma pneumoniae and Legionella spp. Current recommendations for choice of an empirical antibiotic agent are therefore based, not on the assumption of a single etiologic agent indicated by clinical presentation or radiographic appearances, but on age of the patient, severity of illness, the presence of underlying conditions and the range of possible organisms in that patient group. PMID- 11866797 TI - Bacteriologic diagnosis of respiratory tract infections. AB - The use of the Gram stain for determination of the presence of bacteria in respiratory secretions is described. Success of the procedure depends on the use of fresh samples of high cellular quality. The Gram stain can be used to distinguish the presence (among other organisms) of Streptococcus pneumoniae, Haemophilus influenzae, Branhamella catarrhalis enterobacteria and Pseudomonas. This paper also discusses the use of sputum samples for bacteriologic culture and the value of bacterial counts in expectorated sputum, and describes the use of transtracheal aspiration and transthoracic aspiration puncture. Finally, it considers the optimum procedures for the testing of antibiotic susceptibility of presumptive pathogens isolated from the lower respiratory tract. PMID- 11866798 TI - Diagnosis of acute respiratory tract infections: serology and new methods. AB - No test is available that can identify all potential pathogens in acute respiratory tract infections. Each diagnostic test is associated with limitations with respect to sensitivity and/or specificity and/or speed, and thus a combination of tests has to be used. Even so, no etiologic agent is found in 30- 60% of cases. The following methods are recommended for use in the routine laboratory for the diagnosis of infections: (1) Legionella --- direct immunofluorescence test, culture, serology and antigen detection (available only in specialized laboratories); (2) Chlamydia pneumoniae --- micro immunofluorescence test, complement fixation, but not direct antigen detection by immunofluorescence; (3) Mycoplasma pneumoniae - complement fixation and/or particle agglutination or other evaluated methods; (4) Coxiella burnetii --- immunofluorescence test (IgG and IgM) and complement fixation; (5) viruses --- complement fixation or other similar test (ELISAs often lack adequate evaluation). Culture for Chlamydia, Mycoplasma pneumoniae, Coxiella burnetii and viruses should only be performed by very experienced laboratories. Most procedures deliver results only retrospectively or too late. The most promising diagnostic tools for the future are nucleic acid amplification techniques (NAT) or PCR, which could solve many of the problems connected with conventional techniques, but there are enormous contamination problems. Further research and worldwide aid in the development of standardized NAT, especially by industrial companies, is urgently encouraged to improve the laboratory diagnosis of these pneumonias. PMID- 11866799 TI - Non-antibiotic effect of antibiotics. AB - In the present study, we demonstrated the efficacy of low-dose, long-term therapy with erythromycin and an erythromycin-related compound, roxithromycin, in patients with chronic lower respiratory tract disease including diffuse panbronchiolitis (DPB), and investigated the anti-inflammatory action of such drugs. Treatment significantly improved the mean value of respiratory function tests and arterial blood gas analysis, except for PaCO2 in the erythromycin treated group and residual volume/total lung capacity (RV/TLC) and PaCO2 in the roxithromycin-treated group. Bronchoalveolar lavage (BAL) analysis revealed that neutrophils had accumulated in the pre-treatment lavage fluid, compared with that of healthy volunteers, and the levels of neutrophil chemotactic activity (NCA), interleukin-1beta (IL-1beta), interleukin-8 (IL-8) and leukotriene B4 (LTB4) in BAL fluid of the patients were significantly higher than those in healthy volunteers. Macrolide therapy caused a significant reduction in the percentage of neutrophils, NCA and mean IL-1beta, IL-8 and LTB4 concentration in BAL fluid of the patients. The quantitative expression of Mac---1 on peripheral neutrophils significantly increased, compared with that in healthy volunteers, before therapy and significantly decreased after therapy. We also evaluated the in vitro inhibitory effects of macrolides on IL---8 production by vitamin D3-induced human monocytic cell line THP-1 cells when stimulated with lipopolysaccharide and serum. The present study has also provided evidence for T-cell activation in BAL fluid of the patients, and a significantly reduced number of activated T-cells was observed after macrolide therapy. These results indicate that macrolides may act by reducing pulmonary inflammation through reduction of neutrophil accumulation, as a consequence of reduced chemotactic gradient and cytokine production at the inflammatory sites in the lung or of reduced neutrophil adhesion molecules in the circulation. Ultimately the mechanism may involve suppression of neutrophil oxidative and proteolytic products. Lymphocytes are important cellular components of bronchial inflammation, in addition to neutrophils, in this disease, and macrolide antibiotics may also act as an immunosuppressant to inhibit T-cell activation. This is believed to be one reason why macrolides are effective in chronic lower respiratory tract disease. PMID- 11866800 TI - Macrolides as first line therapy in adult lower respiratory tract infections: pros and cons. AB - Certain guidelines exist to facilitate the decision as to which patients with lower respiratory tract infections require antibiotics. The decision as to which antibiotic to use must depend on a detailed knowledge of the disease and the patient's condition. Recent recommendations recognize four categories of patients with community-acquired pneumonia and adopt somewhat different treatment strategies for each. In industrialized countries, macrolides are regarded as either first- or second-choice therapy for community-acquired pneumonias of all grades of severity. PMID- 11866801 TI - Antibacterial treatment of community-acquired pneumonia: the role of therapeutic agents other than the macrolides. AB - The optimal guidelines for the treatment of community-acquired pneumonia are not well established, and the relevant pathogen is often unknown. Initial choice of an antimicrobial agent should depend on the initial severity of the patient's illness, site of acquisition, age, coexisting illness and treatment setting. Few reliable studies have been performed to establish the supremacy of any antibiotic agent in this condition, either for ambulatory or hospital treatment. PMID- 11866802 TI - Final Discussion. PMID- 11866803 TI - LETTER FROM THE EDITOR-IN-CHIEF. PMID- 11866804 TI - Consensus guidelines for appropriate use and evaluation of microbial epidemiologic typing systems. PMID- 11866805 TI - Switch therapy: the theory and practice of early change from parenteral to non parenteral antibiotic administration. AB - OBJECTIVE: The idea behind switch therapy is that antibiotic treatment should be changed from the parenteral to another suitable route (usually oral) as soon as the patient's condition allows. This option is cost-effective in terms of both acquisition costs (oral antibiotics are less expensive than their parenteral counterparts) and indirect costs, and patients may be discharged home sooner. This not only releases hospital beds but is also popular with patients and has other advantages. There are relatively few formal clinical trials, most often using oral third-generation cephalosporins and fluoroquinolones; these agents at present seem the most appropriate to use after parenteral antibiotics have been stopped (usually after 2 to 3 days). Logistic aspects are important, and close collaboration is required between pharmacists, physicians and microbiologists. Further trials are needed in specific patient groups and with other antibiotic regimens to validate the efficacy of switch therapy. PMID- 11866806 TI - Hepatitis C virus genotypes in the liver and serum of patients with chronic hepatitis C. AB - OBJECTIVE: To evaluate the prevalence of hepatitis C virus (HCV) genotypes in a central area of Italy (Umbria); to analyze the correspondence of the genotypes detected in serum and liver samples; to study the relationship between HCV genotypes and severity of liver disease; to test whether co-infection with more than one HCV subtype could be influenced by the source of infection. METHODS: Genotyping by polymerase chain reaction with core-specific primers (Okamoto method) was performed in the serum and liver from 48 consecutive patients with histologically confirmed chronic C hepatitis. RESULTS: HCV genotype 1b was the prevalent strain and was not associated with more severe histologic damage. Data show a very good correspondence between genotypes identified in serum and liver specimens (91%). Mixed infections (with subtypes 1b and 2a) correlated significantly with intravenous drug abuse (p=0.001). CONCLUSION: We confirmed that subtype 1b is prevalent in central Italy. Co-infection with more than one subtype is not rare in intravenous drug abusers. PMID- 11866807 TI - IgG avidity in the serodiagnosis of acute Toxoplasma gondii infection: a multicenter study. AB - OBJECTIVE: To evaluate the validity of the IgG avidity test in the serodiagnosis of acute T. gondii infection; to verify the maturation of IgG avidity during the course of infection; to observe whether the kinetics of IgG maturation could be affected by antibiotic treatment. METHODS: Serial serum samples, collected in three Italian hospitals (Perugia, Treviso and Bologna), from untreated and antibiotic-treated patients with primary toxoplasmic infection, were assayed for IgG avidity, and IgM and IgA positivity. Single serum samples from patients at different stages of infection were assayed for IgG avidity and the results were correlated to the likely stage of infection. RESULTS: The IgG avidity value increased from 3.5% in the first month to 38.7% 1 year from the onset of infection. Antibiotic-treated patients showed significantly different values of IgG avidity at 2 and 4 months after the onset of infection. In single serum samples the IgG avidity values correlated with the likely stage of infection. CONCLUSIONS: The IgG avidity test was confirmed as a useful tool in the serodiagnosis of acute T. gondii infection and could be predictive of the stage of infection. Antibiotic treatment may affect the kinetics of the maturation of IgG avidity. PMID- 11866808 TI - Twice-weekly dapsone for primary prophylaxis against Pneumocystis carinii pneumonia in HIV-1 infection: efficacy, safety and pharmacokinetic data. AB - OBJECTIVES: In this study we evaluated the pharmacokinetics, efficacy and safety of dapsone given 100 mg twice weekly as primary prophylaxis against Pneumocystis carinii pneumonia (PCP) in patients with HIV-1 infection. METHODS: This was a prospective open trial, evaluating a total of 55 HIV-1-infected patients with CD4 cell counts below 200/mm3 and without previous episodes of PCP. Plasma concentrations of dapsone were determined with high-performance liquid chromatography (HPLC). After a mean follow-up of 471 days, the PCP rates per year of observation were 6.79%. Discontinuation of treatment as a result of severe side effects was required in four patients (7.5%). At steady state, mean plasma concentrations 24, 72, 96 and 144 h following the administration of dapsone were 1.46plus minus0.8, 0.28plus minus0.20, 0.30plus minus0.21 and 0.37plus minus0.27 mg/L, respectively. Dapsone plasma levels showed a high interpatient variability. The values for the pharmacokinetic parameters were comparable to those described for healthy volunteers. CONCLUSIONS: The administration of 100 mg twice weekly of dapsone seems appropriate to maintain effective plasma concentrations of the drug and to prevent PCP with good safety in patients with HIV-1-related immunodeficiency. PMID- 11866809 TI - Anti-enolase antibodies partially protective against systemic candidiasis in mice. AB - OBJECTIVE: The evaluation of the effect of protective capacities of antibodies directed against the immunodominant enolase antigen of C. albicans on the survival rate of mice with systemic candidiasis. METHODS: Passive transfer of heterologous rabbit antibodies (IRS) directed against a crude Candida albicans extract and homologous mice anti-enolase antibodies (IMS) in a non-acute but lethal systemic Candida model in mice. RESULTS: Protection could only be demonstrated by repeated administration of homologous as well as heterologous immune sera. The protective effect was not due to non-specific, heat-labile, serum factors, since fractionated anti-cytoplasmic rabbit immunoglobulins also gave a decreased mortality rate. These findings could not be ascribed to enhanced opsonization as observed in in vitro opsonization experiments using peritoneal macrophage monolayers. CONCLUSIONS: Anti-enolase antibodies are partially protective against lethal C. albicans infections. The protection was not due to enhanced initial clearance of viable C. albicans from the internal organs, but was presumably the result of as yet unknown effects of antibodies occurring later in the infectious process. PMID- 11866810 TI - Multiresistant Staphylococcus epidermidis in a neonatal care unit. AB - OBJECTIVE: To study the presence and diversity of types of Staphylococcus epidermidis in the neonatal intensive care unit of a university hospital. METHODS: During a period of 6 weeks, samples were taken from nose, external auditory canal, axilla, groin and umbilicus from consecutively admitted patients. Patients were sampled two times a week for up to 2 weeks. Isolates of S. epidermidis were characterized by antibiogram, plasmid pattern and biotype. RESULTS: Fifteen patients were included. Each patient was sampled in one to four successive surveys, depending on the admission period. A total of 128 isolates of S. epidermidis were obtained and allocated to seven antibiogram types, 36 plasmid types and 14 biotypes. One plasmid type found in 58 isolates (six patients) corresponded with one multiresistant antibiogram type. The number of isolates with these characteristics increased per neonate from the first survey to the fourth. Nineteen isolates from four patients were allocated to a second plasmid type and were of a common antibiogram type. The remaining 34 plasmid types were sporadic. No clear correspondence of biotypes with antibiogram or plasmid types was found. CONCLUSIONS: The present study revealed the increase in colonization of a multiresistant type of S. epidermidis in the compromised patients during admission to the ward. Further studies have to assess whether this type remains persistent in the ward. PMID- 11866811 TI - Toxin A detection on Clostridium difficile colonies from 24-h cultures. AB - OBJECTIVE: Performance of a combined approach for the detection of toxigenic strains in patients suspected of having Clostridium difficile-associated disease was evaluated. METHODS: In this approach, stools were cultured for 24 h on a selective medium supplemented with sodium taurocholate (TCCFA), in anaerobic conditions created with the Martreg Anoxomat system, and toxin A detection was performed directly on C. difficile colonies, by enzyme immunoassay (EIA). This method was compared with three others: cytotoxigenic culture consisting of a 48-h culture on selective medium followed by detection of in vitro cytotoxin production on cell monolayers, fecal cytotoxin detection and fecal toxin A detection by EIA. RESULTS: From 548 stools, 108 yielded a positive culture by at least one of the methods, and 81 isolates were cytotoxin producers. Cultures for 24 h on TCCFA were positive in 106 cases and EIA performed on colonies gave 73 positive results, giving a sensitivity of 90.1% and a specificity of 100%. By comparison, the sensitivity and specificity of cytotoxigenic culture, stool cytotoxin and stool toxin A were respectively 96.2% and 100%, 61.7% and 100%, and 66.7% and 95.9%. CONCLUSIONS: Performing EIA on colonies recovered after 24 h culture allows us to improve the detection of toxigenic strains in patients suspected of having C. difficile-associated disease. PMID- 11866812 TI - Rapid and sensitive detection of Shigella sonnei in feces by the use of an O antigen-specific monoclonal antibody in a combined immunomagnetic separation polymerase chain reaction assay. AB - OBJECTIVE: The aim of the present work was to develop a rapid, specific and highly sensitive diagnostic method for the detection of Shigella sonnei directly from stool samples. DESIGN AND METHODS: An immunomagnetic separation-polymerase chain reaction (IMS-PCR) combined assay for diagnosis of S. sonnei was developed. For this, a monoclonal antibody (Mab) specific for the O-antigen of S. sonnei was coated to magnetic beads for capture, concentration and separation of S. sonnei from feces. Bacterial DNA, was amplified by the PCR with specific primers. The amplified products were developed by dot blot hybridization with a specific alkaline phosphatase-conjugated probe. RESULTS: The purified MASS MAb reacted specifically with the Plesiomonas shigelloides (the same O-antigen as Shigella sonnei) LPS. The primers were specific for invasive Shigella and enteroinvasive Escherichia coli. Only invasive Shigella sonnei strains gave a positive result in the IMS-PCR assay. The detection limit was 10 to 15 c.f.u. CONCLUSIONS: The availability of IMS-PCR assays provides an improved method for the diagnosis of shigellae directly from feces. The assay is rapid, highly sensitive and specific for the detection of Shigella sonnei directly from stool samples. PMID- 11866813 TI - Enterobacterial intergenic consensus sequence polymerase chain reaction as a typing method for Burkholderia (Pseudomonas) cepacia. AB - OBJECTIVE: We developed a rapid polymerase chain reaction (PCR) method, which utilizes primers from an enterobacterial repetitive consensus sequence, to differentiate among strains of Burkholderia cepacia. We then applied this method to isolates from a device-associated pseudoinfection and also compared the discriminative typing potential with that of another PCR-based method. METHODS: A simplified lysis procedure was used to generate DNA substrate for amplification by a PCR method which utilized primer ERIC2 and a reverse complement of ERIC1R. The alternative PCR-based method employed primers which are directed to Bordetella pertussis repetitive sequences. Amplification products were visually assessed after conventional agarose gel electrophoresis and staining. RESULTS: The ERIC-based method appropriately clustered strains for the device-associated pseudo-infection. In addition, there was consistency in the definition of similar or dissimilar strains between the two PCR---based systems. CONCLUSIONS: The ERIC based PCR typing method offers sufficient discriminatory power for it to be used for epidemiologic purposes. PMID- 11866814 TI - Activity in vitro of 22 antimicrobial agents against clinical isolates of Listeria monocytogenes. PMID- 11866815 TI - Multiresistant strain of Listeria monocytogenes in Greece. PMID- 11866816 TI - Fatal septic shock due to Gemella morbillorum in two HIV-positive patients. PMID- 11866817 TI - Postoperative Cryptococcus neoformans endocarditis. PMID- 11866818 TI - Review of publication: Prevalencia de las Infecciones Nosocomiales en los Hospitales Espanoles EPINE 1990--1994. PMID- 11866819 TI - Resurgence of diphtheria in Europe. PMID- 11866821 TI - Antibiotic-resistant bacteria may outweigh their susceptible homologs. PMID- 11866820 TI - Direct cell-to-cell contact mediating upregulation of HIV-1 replication. PMID- 11866822 TI - Hemofiltration: treating and preventing infection. PMID- 11866823 TI - Imported malaria: presentation and outcome of 111 cases. AB - OBJECTIVE: To evaluate the presentation and outcome of imported malaria. PATIENTS AND METHODS: Retrospective charts review of hospitalized patients with smear proven malaria from January 1989 to October 1994. RESULTS: Of a total of 111 cases, 95 were caused by Plasmodium falciparum. Chemoprophylaxis was used in 57% of patients but only 41% were compliant. Parasitemia ranged from 0.1% to 22%. Clinical and biological signs at admission were unspecific. In all cases where both platelets and C-reactive protein were measured, at least one abnormality was noted. More than two WHO gravity criteria were present in 14 cases. Outcome was marked by two deaths and 18 adverse drug reactions. Plasmodium falciparum was associated to only one independent factor in multivariate analysis: symptom onset less than 30 days after return. Chemoprophylaxis use did not modify clinical presentation, mean parasitemia or outcome. CONCLUSIONS: P. falciparum is the most usual cause of imported malaria in France. Normal platelet count and C-reactive protein value probably exclude the diagnosis of malaria in febrile travelers. PMID- 11866824 TI - Meropenem in cystic fibrosis patients infected with resistant Pseudomonas aeruginosa or Burkholderia cepacia and with hypersensitivity to beta-lactam antibiotics. AB - OBJECTIVE: To assess the efficacy and safety of meropenem, administered on a compassionate basis to 62 cystic fibrosis (CF) patients (age: 24plus minus6 years) with hypersensitivity reactions to beta-lactam antibiotics and/or infection by bacteria resistant to other antibiotics. METHODS: Fifty-seven patients were chronically infected with Pseudomonas aeruginosa and 5 with Burkholderia cepacia. In total, 124 courses (1 to 6/patient) of meropenem, 2 g three times a day by intravenous infusion (10 to 15 min) for 14 days, were administered. RESULTS: During treatment for P. aeruginosa the mean increase in pulmonary function (as a percentage of the predictive values) was 5.6% for FEV1 (forced expiratory volume in the first second) and 8.6% for FVC (forced vital capacity). C-reactive protein and erythrocyte sedimentation rate (ESR) and leukocyte count decreased significantly. In courses administered for chronic infection with B. cepacia the post treatment FEV1 and FVC values were higher than the pre-treatment values, and all the inflammatory parameters decreased. The geometric means of minimal inhibitory concentration (MICs) (microg/mL) for P. aeruginosa (B. cepacia) were: tobramycin 6 (59), ciprofloxacin 1.2 (9.7), piperacillin 49 (16.3), ceftazidime 26 (23), aztreonam 26 (35), imipenem 6.4 (not determined) and meropenem 5.1 (4.8). No statistically significant increase in the MICs of meropenem for either pathogen occurred during therapy. Of the 124 courses, 115 were tolerated without any clinical complaint. The following side effects were observed: nausea (0.8%), itching (4%), rash (3.2%), drug fever (1.6%). CONCLUSIONS: Meropenem proved to be a valuable drug in the treatment of CF patients with chronic pulmonary infection with multiresistant P. aeruginosa and B. cepacia and with hypersensitivity reactions to other beta-lactam drugs. PMID- 11866825 TI - Determination of in vitro postantibiotic effects in Staphylococcus aureus and Escherichia coli by [3H]thymidine incorporation. AB - Postantibiotic effects (PAE) and control-related effective regrowth time (CERT) of dicloxacillin, vancomycin, rifampin and gentamicin in Staphylococcus aureus and imipenem, gentamicin, tobramycin, doxycycline and rifampin in Escherichia coli were measured by standard viability counting and [3H]thymidine incorporation. For PAE determination, the two methods correlated well; r2=0.821 for S. aureus and r2=0.939 for E. coli. For viable counts below the detection limits of 105 to 106 log10 CFU/mL, the PAE was overestimated by the [3H]thymidine method. Quantitation of CERT by both methods showed a good correlation, r2=0.867 for S. aureus and r2=0.997 for E. coli. Measuring [3H]thymidine incorporation in bacteria is a novel alternative method for the determination of PAE and CERT. PMID- 11866826 TI - Persistent parvovirus B19 infections with different clinical outcomes in renal transplant recipients: diagnostic relevance of polymerase chain reaction (PCR) and of quantification of B19 DNA in sera. AB - OBJECTIVE: To study parvovirus B19 infection in immunocompromised subjects such as renal transplantation recipients. METHODS: Two cases of B19 infection in renal transplant recipients have been included in the study. The outcome of the infection has been studied by both serologic and virologic methods. A monitoring of the DNAemia was done by a nested PCR in endpoint titration assays. RESULTS: In one patient with severe anemia an acute B19 infection was diagnosed by PCR 26 days after the transplant; a high level of DNAemia persisted until an intravenous immunoglobulin treatment. Then a sharp decrease of the DNAemia was shown, without full clearance of B19 virus. In a lymphocyte suspension from the organ donor, B19 DNA was detected. In the other patient, who recovered spontaneously from anemia, a persistent B19 infection was demonstrated at day 106 after transplantation and was still demonstrable after 470 days. CONCLUSIONS: A high level of B19 DNAemia was associated with symptomatic infection, with severe anemia, whereas low-level DNAemia was long-lasting in asymptomatic subjects with impaired immunologic responses. The endpoint titration assay by nested PCR was very useful for the monitoring of B19 infection, particularly following the therapeutic intravenous immunoglobulin administration. PMID- 11866827 TI - Detection of enteroviral infection in myocardial tissues by polymerase chain reaction (PCR). AB - OBJECTIVE: To determine if enteroviral infection is linked to myocarditis and dilated cardiomyopathy. Enteroviruses, especially coxsackieviruses, appear to be the most common agents of viral myocarditis. METHODS: We collected 53 endomyocardial biopsies and two autopsy specimens from 41 patients affected by myocarditis or dilated cardiomyopathy. The patients were diagnosed clinically, hemodynamically, virologically and histologically (Dallas classification). We tested for the presence of enteroviral sequences by PCR, using 5prime prime or minute non-coding (coxsackievirus B3, CB3, map position 450--474, 584--603) derived primers. Specificity was confirmed using the Southern blot. We used a fraction of CB3 acutely infected mouse myocardial tissue as a control. RESULTS: We detected enteroviral sequences in four patients with active myocarditis, borderline myocarditis or cardiomyopathy. The patient with active myocarditis had shown neutralizing antibodies in serologic analysis for coxsackievirus B3 and B5. CONCLUSIONS: The data support a weak link of enteroviral infection to human myocarditis and dilated cardiomyopathy, at least when using a PCR assay on biopsies. PMID- 11866828 TI - Evaluation of the Rapid ID 32A system for the identification of the Bacteroides fragilis group. AB - OBJECTIVE: To evaluate the use of a rapid identification system, Rapid ID 32A (bioMerieux), for the identification of clinically important species in the B. fragilis group. METHODS: The use of Rapid ID 32A was validated on 249 clinical isolates, all of which were tested by conventional techniques, and in selected instances API 20A. Rapid ID 32A (and API 20A as appropriate) was then applied in a central laboratory to the identification of 1289 B. fragilis group clinical isolates from 22 laboratories in 15 European countries. RESULTS: Improvements in the initial database permitted the accurate identification of isolates of B. fragilis, B. thetaiotaomicron and B. vulgatus, but further tests, especially for catalase production, were required to distinguish between B. ovatus and B. uniformis, while an identification of B. distasonis could be accepted only after careful review of results. There were too few isolates of B. caccae, B. merdae and B. stercoris for us to reach satisfactory conclusions, but further tests are clearly necessary. CONCLUSIONS: The study emphasizes the importance of including sufficient numbers of isolates of different species in the validation of identification methods. Rapid ID 32A is a reliable system for the identification of the common species in the B. fragilis group, especially B. fragilis and B. thetaiotaomicron. PMID- 11866829 TI - Use of the ligase chain reaction on urine of men and their female sexual partners for detection of genital Chlamydia trachomatis infection. AB - OBJECTIVE: The purpose of the present study was to evaluate an in vitro DNA amplification assay named the ligase chain reaction (LCR) for the detection of Chlamydia trachomatis cryptic plasmid DNA in urine from men and women, in comparison with urethral swab culture in men and cervical swab culture in women. METHODS: 591 patients (394 men with urethritis and 197 female sex partners) attending a center for sexually transmitted diseases in northern Italy between January 1994 and January 1995 were enrolled in this study. A cervical swab was collected from women and a urethral swab from men for standard tissue cell culture. From each patient 20 mL of the first stream of the urine (FVU), taken at least 2 h after the last urination, were collected for LCR analysis. Discrepant results were further analyzed by direct fluorescence and a LCR with alternative primers. RESULTS: In men the prevalence of C. trachomatis infection by urethral culture was 13.45% and, after resolution of discordant results, the LCR method performed on FVU showed a sensitivity, specificity, positive predictive value and negative predictive value of 89.4%, 100%, 100% and 98.2%, respectively; the sensitivity of tissue cell culture was 92.8%. In female sex partners, the prevalence of C. trachomatis infection by cervical culture was 3.04%; LCR detected eight true positive samples, two more than tissue cell culture, and no false-negative results. CONCLUSION: LCR analysis of FVU is a rapid, non-invasive technique and represents a good alternative to tissue cell culture. Further study is needed to investigate possible LCR inhibitors present in urine samples. PMID- 11866830 TI - Direct detection of Mycobacterium tuberculosis in respiratory samples from patients in Scandinavia by polymerase chain reaction. AB - OBJECTIVE: To investigate the use of DNA amplification by the polymerase chain reaction (PCR) for the detection of Mycobacterium tuberculosis directly in human respiratory specimens. METHODS: The PCR assay employed was the Amplicor M. tuberculosis Test (Roche Diagnostics, Switzerland), which uses the 16S rDNA as the target template. Nine hundred and sixty samples from 741 patients in two clinical microbiology laboratories in Norway and Sweden were processed by routine culture analysis and PCR. RESULTS: Of the 56 specimens containing cultivatable M. tuberculosis, 49 (87.5%) were detected by PCR. Among the 904 culture-negative specimens, 897 samples were negative also by PCR and seven (0.8%) were positive by PCR. In comparison with culture, the sensitivity, specificity, and positive and negative predictive values of PCR were 91.7%, 99.6%, 94.2% and 99.4% for laboratory 1 and 80.0%, 98.7%, 76.2% and 99.0% for laboratory 2, respectively. For both laboratories combined the values were 87.5%, 99.2%, 87.5% and 99.2%. CONCLUSIONS: These results indicate that multiple (two or three) respiratory samples from each patients should be tested, to allow sufficient accuracy in detecting M. tuberculosis in the specimens. Still, the labor-intensive format of this test necessitates strong clinical indications and patient prioritization to provide a service feasible within the current limits of routine laboratories. PMID- 11866831 TI - An overview of the hepatitis viruses. PMID- 11866832 TI - Streptococcal perianal disease in two healthy adult women. PMID- 11866833 TI - Clinical impact of a 5-day versus 7-day blood culture incubation period. PMID- 11866834 TI - Asymptomatic carriage of Neisseria meningitidis in a randomly sampled population. Serogroup, serotype and subtype distribution and associated risk factors. PMID- 11866836 TI - 1997 ESCMID Awards. PMID- 11866837 TI - New molecular techniques and the study of infection. PMID- 11866835 TI - Endogenous endophthalmitis due to Streptococcus agalactiae: case report and review. PMID- 11866838 TI - Susceptibility of coagulase-negative staphylococci to teicoplanin. PMID- 11866839 TI - Novel human herpesviruses (human herpesviruses 6, 7 and 8). AB - The number of members in the family Herpesviridae has increased in the last 10 years due to the description of three novel human herpesviruses: human herpesvirus 6 (HHV-6) in 1986, human herpesvirus 7 (HHV-7) in 1990, and human herpesvirus 8 (HHV-8), also known as Kaposi's sarcoma-associated herpesvirus (KSHV), in 1994. HHV-6 and HHV-7 were first isolated from blood lymphocyte cultures, while HHV-8 was identified following a specific molecular biology approach in the search for the etiologic agent of Kaposi's sarcoma. The three viruses are lymphotropic, T-cells being the targets of HHV-6 and HHV-7, and B cells being probably those of HHV-8. The ability to be propagated in cell cultures in vitro differs according to the virus concerned: this can be done readily with HHV-6, with more difficulties in the case of HHV-7, and has not yet been achieved in the case of HHV-8. Human infection with HHV-6 and HHV-7 is ubiquitous, widespread and acquired early in life. HHV-8 epidemiology is still unclear, and there are two hypotheses: a restricted dissemination in the general population like herpes simplex virus type 2, or a widespread infection like all other human herpesviruses. The polymerase chain reaction is the common method for the detection of infection using specific primers and probes for HHV-6, HHV-7 and HHV-8 respectively. Serologic assays are only available for HHV-6 and HHV-7, with limitations being due, in particular, to possible cross-reactions with cytomegalovirus. HHV-6 is the causative agent of exanthem subitum (sixth disease). Its role as an opportunistic agent and immune dysfunction inducer is debated and currently under investigation. The pathogenic role of HHV-7 seems to be modest, with one case of exanthem subitum reported so far. HHV-8 is strongly associated with three diseases: Kaposi's sarcoma, Castleman's disease and body cavity-based lymphomas. The therapy against these novel viruses has to be considered in the future. PMID- 11866840 TI - Endemic meningococcal disease in Cerdanyola, Spain, 1987--93: molecular epidemiology of the isolates of Neisseria meningitidis. AB - OBJECTIVE: To establish the relationships between 30 Neisseria meningitidis strains isolated in Cerdanyola (Spain) from 30 out of 36 sporadic cases of meningococcal disease (MD) during 1987--93 and their spread in this population by multilocus enzyme electrophoresis (MEE) and by pulsed-field gel electrophoresis (PFGE), and to evaluate the usefulness of PFGE versus serologic typing methods and MEE as an alternative epidemiologic marker to study meningococcal infection. METHODS: Serotyping, electrophoretic mobility of seven isoenzymes determined by MEE and chromosomal DNA macrorestriction with NheI resolved by PFGE were analyzed. RESULTS: Of these 30 strains, 25 were serogroup B and the remaining five were serogroup C, with the 4:P1.15 and the 2b:NT as the most common antigenic phenotypes, respectively. There were 13 electrophoretic types (ETs) by MEE, with 14 isolates showing an identical ET, 8. Sixteen pulse types (PTs) were generated by PFGE. The 14 ET 8 isolates were clustered into six PTs, A1, A2, A4, A5, A6 and A8. However, by combining both methods, 19 genetically distinct groups were obtained. Eleven of these groups (20 serogroup B strains) and two of these (four serogroup C strains) were genetically related. CONCLUSIONS: We conclude that, according to the clonal population structure, these 30 N. meningitidis strains are heterogeneous although a great number are related. Moreover, PFGE is a useful method to establish clonal structure in N. meningitidis strains under endemic conditions. Finer discrimination of these strains was achieved by combining both MEE and PFGE methods. PMID- 11866841 TI - Stomatococcus mucilaginosus septicemia in leukemic patients. AB - OBJECTIVE: To report an unexpectedly high number of cases of septicemia with Stomatococcus mucilaginosus, and try to identify predisposing factors. METHODS: All blood cultures obtained during 1991--93 from patients treated at the hematologic ward were bacteriologically identified. The medical records of patients with S. mucilaginosus-positive blood cultures were retrospectively reviewed and evaluated. The antibiotic susceptibility pattern and restriction fragment length polymorphism (RFLP) of S. mucilaginosus were tested. RESULTS: S. mucilaginosus blood isolates from patients with hematologic malignancies were found to be as common as isolates of Staphylococcus aureus. Eleven patients with myelogenous leukemia and isolation of S. mucilaginosus from the blood are reported on. One patient had concomitant meningitis. All patients were neutropenic and most had oral mucositis and had been given ciprofloxacin prophylaxis. S. mucilaginosus isolates from these patients were resistant to ciprofloxacin in contrast to isolates from patients who had received other prophylactic regimens and seven isolates found in healthy individuals not recently treated with antibiotics. The resistant S. mucilaginosus were found to be of diverse genetic origin as determined by RFLP. CONCLUSIONS: The appearance of resistant strains during ciprofloxacin prophylaxis may be a predisposing factor for S. mucilaginosus septicemia. There was no evidence of a nosocomial spread of S. mucilaginosus strains. PMID- 11866842 TI - Pharmacokinetics of once-daily amikacin in pediatric patients. AB - OBJECTIVE: To study the pharmacokinetic parameters of a once-daily regimen of amikacin (15 mg/kg) in association with other antimicrobial agents in 35 children with severe Gram-negative infections. METHODS: A Bayesian approach was developed to optimize the amikacin regimen. The predictive performance was assessed by computing bias and precision. Each patient was evaluated for toxicity after 5 days of treatment. RESULTS: Peak amikacin concentrations on days 2 and 5 of therapy averaged 31.3 plus minus 9.0 mg/L and 32.4 plus minus 7.4 mg/L, respectively. To achieve peak serum concentrations between 30 and 40 mg/L, individualized dosage was necessary in 19 of 35 children. The pharmacokinetic parameters showed large interindividual variations, with a mean half-life of 2 h and a mean volume of distribution of 0.36 L/kg. No nephrotoxicity was observed in any of the children. After individualization of dosage on the basis of one measurement of peak concentration, no significant differences were observed between predicted and subsequently measured amikacin concentrations. CONCLUSIONS: Once-daily dosage of amikacin (15 mg/kg) is well tolerated in pediatric patients; however, a loading dose of 20 mg/kg is recommended to achieve a therapeutic peak value between 30 and 40 mg/L. Initial serum monitoring is essential in a population such as children, with wide interpatient variability. Using the Bayesian approach, the amikacin regimen in children can then be predicted with minimal bias and good precision. PMID- 11866843 TI - Surveillance of intestinal colonization and of infection by vancomycin-resistant enterococci in hospitalized cancer patients. AB - OBJECTIVE: To study epidemiologic features of and risk factors for intestinal colonization and infection by vancomycin-resistant enterococci (VRE) in cancer patients. METHODS: During a 41-month period, over 7600 fecal samples and all samples from sterile sites from hospitalized cancer patients were screened for VRE. Species were identified and isolates analyzed by pulsed-field gel electrophoresis (PFGE) of SmaI DNA restriction fragments. Antibiotic resistance was characterized by MIC determinations, and polymerase chain reaction for vanA, vanB, and vanC1 genes. Plasmid contents were analyzed before and after PstI and HindIII restriction, and by Southern hybridization with a vanA probe. Two case control studies were performed to identify risk factors for colonization or infection by VRE, respectively. RESULTS: Eighty-two isolates were recovered from 81 patients. Most (72%) isolates were Enterococcus faecium VanA/vanA, with 37 different PFGE types, each of which was found in only one to four patients, except for type P1, which was found in 20 patients hospitalized over a 3-month period in the pediatric wards. Plasmid analysis suggested that only two types of plasmid were carrying gene vanA, as part of a transposon related to transposon Tn 1546 from reference strain E. faecium BM4147. Seventy-seven patients were colonized during the study period. Six of them became infected. Four patients were infected but not colonized. Only one patient died during the course of infection, but intestinal colonization persisted for months in the survivors. Case-control analysis revealed that cephalosporin treatment was a significant risk factor for colonization. No significant risk factor for infection was found in colonized patients. CONCLUSION: Colonization by VRE was mostly endemic and the colonized patients were not often infected. However, when clustered cases of colonization occurred, they were then associated with an increased rate of infection. PMID- 11866844 TI - Routine use of CHROMagar Candida medium for presumptive identification of Candida yeast species and detection of mixed fungal populations. AB - OBJECTIVE: To assess the value of the new differential culture medium CHROMagar Candida for routine investigation of clinical specimens. METHODS: During a whole year, 6150 clinical samples were plated on CHROMagar Candida medium. After incubation, the green colonies were considered to be Candida albicans. The colonies of other colors were identified using Bichrolatex-krusei, or by their assimilation pattern on ID 32C test strips and their morphology on rice cream agar-Tween. RESULTS: Among the 6150 clinical samples, 1643 were positive for fungi. Aspergillus fumigatus and Geotrichum sp. were the predominant filamentous fungi isolated. Candida albicans was the most common species isolated (1274 of the positive samples; 77.5%), and Candida glabrata was the second most common yeast isolated (174 positive samples; 10.6%). Other yeast species were detected at lower frequencies, mainly Candida tropicalis (3.8%), Candida krusei (2.7%), Saccharomyces cerevisiae (2.7%) and Candida kefyr (2.3%), and 16 samples revealed a lipophilic species, Malassezia furfur. Mixed fungal populations accounted for 14.7% of the positive samples. Two or more yeast species were detected in 206 of the 242 specimens containing mixed fungal populations, and five yeast species were detected in one sample. Additionally, we did not observe significant differences in the isolation of yeasts or filamentous fungi from the 366 samples simultaneously plated on CHROMagar Candida and Sabouraud dextrose agar. Close agreement between the two culture media was observed for 89.9% of these samples. CONCLUSIONS: CHROMagar Candida medium was shown to be extremely helpful in a routine clinical mycology service, facilitating the detection of mixed cultures of yeasts and allowing direct identification of C. albicans, as well as rapid presumptive identification of the other yeasts: C. glabrata, C. tropicalis, C. krusei and S. cerevisiae. This chromogenic medium thus appears to be suitable as a primary culture medium, particularly for the mycologic surveillance of immunocompromised patients. PMID- 11866845 TI - Disk diffusion, agar dilution and the E-test for susceptibility testing of Corynebacterium jeikeium. AB - OBJECTIVE: The susceptibilities to penicillin, tetracycline, erythromycin, gentamicin, vancomycin and teicoplanin of 58 strains of Corynebacterium jeikeium were assessed by disk diffusion and agar dilution reference methods. METHODS: Zone sizes and minimal inhibitory concentrations (MIC) by agar dilution were interpreted using the ranges in the NCCLS tables for organisms other than Haemophilus, Neisseria gonorrhoeae, and Streptococcus pneumoniae. RESULTS: By agar dilution, 14%, 88%, 17% and 26% of the 58 isolates were susceptible to penicillin, tetracycline, erythromycin, and gentamicin, respectively. Using the breakpoints for Listeria monocytogenes, all strains showed concordant results for penicillin by disk diffusion. Discrepancies in the interpretative categories by disk diffusion were found in four cases (two very major and two minor) for tetracycline, in nine (two very major, two major, and five minor) for erythromycin, and in 1 case (very major) for gentamicin. All 58 strains were susceptible to vancomycin and teicoplanin by agar dilution and disk diffusion. The overall agreement of interpretative disk diffusion for all six antibiotics was 95.9%. In addition, all strains were susceptible to both glycopeptides by E test. However, for vancomycin the MIC results in 58.6% were two log2 dilutions and in 1.7% more than two log2 dilutions higher by E-test than by agar dilution, whereas for teicoplanin agreement within one log2 dilution was 100%. CONCLUSIONS: Further evaluation of methodologies of disk diffusion is required to obtain a better agreement for erythromycin and tetracycline. The criteria of the NCCLS for interpretation of disk diffusion are adequate for susceptibility testing of C. jeikeium to penicillin, gentamicin, vancomycin and teicoplanin. PMID- 11866846 TI - Comparison of A60 and three glycolipid antigens in an ELISA test for tuberculosis. AB - OBJECTIVES: To compare the diagnostic usefulness in tuberculosis of the serodiagnostic enzyme-linked immunosorbent assay (ELISA) kit A60 (Anda Biologicals, Strasbourg, France) and of our domestic ELISA based on three purified cell wall glycolipid antigens. METHODS: The presence and concentrations of IgG and IgM anti-A60 antibodies and anti-LOS, anti-DAT and anti-PGLTb1 antibodies against the glycolipid antigens were determined by ELISA in 50 HIV seronegative and 46 HIV-seropositive patients, with documented active tuberculosis. The specificity of these ELISAs was determined with use of sera from 50 healthy blood donors, 29 patients with non-mycobacterial pulmonary diseases and 24 HIV-positive patients with disseminated Mycobacterium avium infection. RESULTS: With a calculated cut-off for each antigen and immunoglobulin that gave a specificity higher than or equal to 98%, the cumulative ELISA results showed that only 36.5% of the patients with tuberculosis had a positive response in the A60 test, as compared with 84.4% who showed a response to the three glycolipid antigens (p<0.001). This striking difference persisted when the cumulative sensitivities were calculated according to the HIV status of the patients and the localization of tuberculosis. The anti-A60 antibody (IgG and IgM) levels and the degree of sensitivity of the ELISA for detection of A60 antigen were always lower in HIV-positive patients with pulmonary and extrapulmonary tuberculosis than in HIV-negative patients with tuberculosis. The sensitivity of A60 ELISA was further decreased in HIV-positive patients with low CD4+ lymphocytes counts, in contrast to the results with the three glycolipid antigens. CONCLUSIONS: These results show the limitations of the A60 ELISA, and confirm the potencies of the glycolipid antigens in serodiagnosis of tuberculosis in HIV-positive and HIV-negative patients. PMID- 11866847 TI - Immunology and management of sepsis. PMID- 11866848 TI - Disseminated adenovirus 20 infection with pancreatitis in a patient with AIDS. PMID- 11866849 TI - Bacterial endocarditis due to Lactococcus lactis subsp. cremoris: case report. PMID- 11866850 TI - Observation of microcolonies on Middlebrook 7H11 medium for rapid detection of Mycobacterium tuberculosis and the Mycobacterium avium complex in clinical samples. PMID- 11866852 TI - Erratum from Volume 2 Number 1, page 5. Consensus guidelines for appropriate use and evaluation of microbial epidemiologic typing systems. PMID- 11866851 TI - Francisella tularensis isolation from various clinical specimens. PMID- 11866854 TI - Indexes to Volume 2. PMID- 11866855 TI - Why a National Committee? PMID- 11866856 TI - Overview of the Comite de l'Antibiogramme de la Societe Francaise de Microbiologie. PMID- 11866858 TI - Technical recommendations for in vitro susceptibility testing. PMID- 11866857 TI - Definition and determination of in vitro antibiotic susceptibility breakpoints for bacteria. PMID- 11866859 TI - Interpretive reading of in vitro antibiotic susceptibility tests (the antibiogramme). PMID- 11866860 TI - Detection of extended-spectrum plasmid-mediated beta-lactamases by disk diffusion. PMID- 11866862 TI - Statement 1996 CA-SFM Zone sizes and MIC breakpoints for non-fastidious organisms. PMID- 11866861 TI - Definition of the Clinical Antibacterial Spectrum of Activity. PMID- 11866863 TI - Assessing health-related quality of life in patients with inflammatory bowel disease, in Crete, Greece. AB - BACKGROUND: Health Related Quality of Life (HRQoL) is an important outcome measure in Inflammatory Bowel Disease (IBD). The aim of our study was to assess HRQoL in a population of 135 Greek patients with IBD. METHODS: A cohort of 135 patients with IBD, 81 with ulcerative colitis (UC) and 54 with Crohn's disease (CD) were enrolled in our study. Demographic and disease-related data were recorded. HRQoL was assessed by a disease-specific and a generic questionnaire, IBDQ and SF-36, respectively. Disease activity was assessed by Harvey-Bradshaw Index and the Colitis Activity Index for CD and UC patients, respectively. RESULTS: Among all variables recorded in our study, only disease activity had a significant effect on HRQoL. Patients with active disease scored significantly lower on both IBDQ and SF-36 when compared to those in remission. Only two among the four IBDQ dimensions, bowel and systemic, had significant ability in distinguishing best patients in remission from those with active disease. CONCLUSIONS: IBD has a negative impact on HRQoL. Patients with active disease are more impaired than patients in remission. In our population of patients bowel and systemic dimensions had a predominant value in patients' perception of quality of life. Patients in our study using the same instrument scored higher than previously reported. PMID- 11866864 TI - A prospective, cross-sectional study of anaemia and peripheral iron status in antiretroviral naive, HIV-1 infected children in Cape Town, South Africa. AB - BACKGROUND: Anaemia is a common manifestation of paediatric HIV infection. Although there are many causes, anaemia of chronic diseases is the most frequent type. In poor countries iron deficiency is widespread. It is probable that many HIV-infected children in these countries are also iron deficient. This study describes the relationship between paediatric HIV infection and anaemia, and documents the peripheral iron status of antiretroviral naive, HIV-infected children. METHODS: Sixty children were evaluated prospectively. Investigations included CD4+ count, haemoglobin concentration (Hb), red blood cell (RBC) morphology, and iron studies. RESULTS: Anaemia was present in 73% of children. Compared to mild HIV infection, median Hb was lower in children with moderate clinical infection (104 g/L v 112 g/L, p = 0.04) and severe clinical infection (96 g/L v 112 g/L, p = 0.006), and more children with severe infection were anaemic (92% v 58%, 0.04). There was a significant relationship between immunological status and Hb. 68% had abnormal RBC morphology. Significantly more children with moderate and severe disease, and severe immunosuppression had abnormal RBC morphology. 52% were iron-depleted, 20% had iron-deficient erythropoiesis and 18% iron deficiency anaemia (IDA). 16% (7/44) of anaemic children had microcytosis and hypochromia. Median soluble transferrin receptor concentration was significantly higher in those with microcytic hypochromic anaemia (42.0 nmol/L v 30.0 nmol/L, p = 0.008). CONCLUSIONS: Both the proportion of anaemic children and the median Hb were associated with disease status. Iron depletion and IDA are major problems in HIV-infected children in South Africa. PMID- 11866865 TI - Schonlein-Henoch purpura during pregnancy with successful outcome for mother and newborn. AB - BACKGROUND: Schonlein-Henoch purpura is a systemic vasculitis that affects vessels of a small caliber and rarely reported in the literature. CASE PRESENTATION: We report on a 35-year-old woman who developed palpable purpura with necrotizing cutaneous lesions on the lower limbs at 27 weeks of gestation. She also complained of epigastric pain and arthralgias. Histologic examination of a skin biopsy showed leukocytoclastic vasculitis with intravascular fibrin thrombi. The direct immunofluorescence analysis evidenced vascular deposits of IgA and C3 in the upper and mid-dermis. These findings were consistent with Schonlein-Henoch purpura. There was no evidence of renal involvement or placental dysfunction. The patient was treated with low-dose oral corticosteroids and a healthy infant was delivered by cesarean section. Examination of the placenta and the navel string disclosed no signs of vasculitis or infarction. CONCLUSION: Schonlein-Henoch purpura is rarely reported in pregnancy. Treatment with orally administered corticosteroids may lead to a beneficial outcome for mother and newborn. PMID- 11866866 TI - Diagnostic criteria for diabetes revisited: making use of combined criteria. AB - BACKGROUND: Existing cut-offs for fasting plasma glucose (FPG) and post-load glucose (2hPG) criteria are not equivalent in the diagnosis of diabetes and glucose intolerance. Adjusting cut-offs of single measurements have not helped so we undertook this project to see if they could be complementary. METHODS: We performed oral glucose tolerance tests and mean levels of hemoglobin A1c (HbA1c) measurements on 43 patients referred to a diabetes clinic for possible diabetes. Results of single and combined use of the FPG and 2hPG criteria were evaluated against the levels of HbA1c and results re-interpreted in the light of existing reports in the literature. RESULTS: Our results confirm that the FPG and the 2hPG, being specific and sensitive respectively for the presence of glucose intolerance or diabetes, are not equivalent. They are shown to be indeed complementary and a re-definition of diagnostic criteria based on their combined use is proposed. CONCLUSIONS: We conclude that altering single measurement cut offs for the diagnosis of diabetes and altered glucose tolerance will not result in better outcomes. We present the case for a combined criteria in the diagnosis and definition of diabetes with a FPG>/=7 mmol/L AND 2-hour glucose >/=7.8 mmol/L being used to define diabetes while a FPG<7 mmol/L AND 2-hour glucose <7.8 mmol/L being used to define normality. Discordant values will define impaired glucose tolerance (IGT). This proposal requires prospective evaluation in a large cohort. PMID- 11866867 TI - Direct assessment and diminished production of morphine stimulated NO by diabetic endothelium from saphenous vein. AB - AIM: To directly measure in real time basal and stimulated levels of NO released from human saphenous vein endothelium and to quantify the expression of the mu opiate receptor, which has been linked with NO release. METHODS: Saphenous vein segments from patients with type 2 diabetes (n=12) and patients without diabetes (n=8) were obtained. The release of NO was measured directly from the endothelium using a NO-specific amperometric probe. N(Omega)-nitro-L-arginine methyl ester (L NAME, 0.1 mmol/L), a NO synthase (NOS) inhibitor, or morphine (1 mumol/L), a stimulant, was administered and the measurements were repeated. Values were reported relative to the mean initial measurement of NO release from diabetic endothelium, which was defined as the relative zero level of NO release. A RT-PCR was then performed on the endothelium to measure mu opiate receptor expression. RESULTS: Diabetic patients (n=12) showed a relative and significantly diminished basal level of released NO, (0.049+/-0.012) nmol/L, compared with non diabetic patients (n=8), (0.42+/-0.12) nmol/L (P<0.05). Application of L-NAME to nonstimulated tissues resulted in no change in NO release from the diabetic group and a decrease in NO release of (0.21+/-0.09) nmol/L from the non diabetic group (P<0.05). Morphine stimulation of the diabetic endothelium resulted in a lower peak and shorter duration of NO release compared to the non-diabetic tissue, (21+/-6) nmol/L vs (38+/-4) nmol/L and (7.3+/-1.4) min vs (12.2+/-2.2) min, respectively (P<0.01). Lastly, evaluation of the mu opiate receptor expression was found to be diminished in the diabetics by 59.1 %. CONCLUSION: Maturity-onset diabetes attenuates both the constitutive basal and morphine stimulated NO release from human saphenous vein endothelium. In this study, after NOS inhibition, the actual basal NO release in diabetes was negligible. One explanation for the impaired capacity of diabetic endothelium to release NO was the diminished mu opiate receptor that was seen in diabetic endothelium. PMID- 11866868 TI - Determination of arterial baroreflex-blood pressure control in conscious rats. AB - AIM: To study the determination of arterial baroreflex-blood pressure control (ABR-BP) in conscious rats. METHODS: Blood pressure was continuously recorded with a computerized system in conscious freely moving rats. The principle of ABR BP measurement is to compare the pressor response to a vasoactive drug (angiotensin II) before and after the interruption of this reflex. RESULTS: (1) ABR-BP values revealed by angiotensin II were closely correlated with those by phenylephrine. Doses of angiotensin II did not influence the results within certain range. (2) ABR-BP was well correlated with arterial baroreflex-heart period control (ABR-HP). (3) Anesthesia inhibited ABR-BP. There existed a circadian variation of ABR-BP in WKY rats. (4) Blood pressure variability was closely related to ABR-BP, but not to ABR-HP. (5) ABR-BP was impaired in hypertensive rats. CONCLUSION: ABR-BP is an important parameter to reflect the function of ABR. The present work makes it possible to determine ABR-BP in conscious rats. ABR-BP plays an important role in maintaining blood pressure stability and it is impaired in hypertension. PMID- 11866869 TI - Characterization of transient outward K+ current and ultra-rapid delayed rectifier K+ current in isolated human atrial myocytes from patients with congestive heart failure. AB - AIM: To study the properties of transient outward K+ current (Ito) and ultra rapid delayed rectifier K+ current (IKur) in isolated human atrial myocytes from patients with congestive heart failure (CHF). METHODS: Single cells were isolated from CHF patients with collagenase and protease. Ito and IKur were recorded using whole cell patch-clamp technique. RESULTS: The activation and inactivation of I(to) were voltage-dependent and time-dependent. The half-activation and half inactivation voltage were (15 +/- 12) mV and (-45 +/- 4) mV respectively. When membrane potential went up from -40 mV to +60 mV, the activation time constant means decreased from (6.9 +/- 2.3) ms to (1.40 +/- 0.20) ms, while the inactivation time constant means decreased from (69 +/- 17) ms to (21 +/- 14) ms. Otherwise, the mean reactivation time constants was (125 +/- 65) ms when the membrane potential was held at -80 mV, but the recovery was not complete during the interval observed. Ito showed less frequency-dependent reduction at test frequency between 0.2-2 Hz. Compared with Ito, the activation of IKur only showed voltage-dependence, without time-dependence. Its mean current densities was (3.4 +/- 0.7) pA/pF when test potential was +60 mV. The half activation voltage of IKur was (23 +/- 14) mV. No clear frequency-dependence was observed at the same frequency range of Ito either. CONCLUSION: I(to) and IKur are important outward potassium channel currents in isolated human atrial myocytes from CHF patients and they have different kinetic properties. PMID- 11866870 TI - A new model for random screening inhibitors of vascular endothelial growth factor receptor 1 kinase. AB - AIM: To establish a 96-well plate based kinase assay using a recombinant vascular endothelial growth factor (VEGF) receptor 1 kinase domain protein. METHODS: A human VEGF receptor 1 kinase domain protein was expressed in E coli, and its activity was monitored by its ability of phosphorylating the polyE4Y substrate coated on the walls of 96-well plates with antibody recognition and a colorimetric readout. A random screening of a sample organic compound library was carried out, and the hits were characterized with a transformed cell line stably expressing VEGF receptor 1 protein. RESULTS: An efficient E coli expression system for human VEGF receptor 1 kinase domain protein was constructed, and the purified recombinant protein was used to establish a practical screening assay for kinase inhibitors in vitro. Two thousand eight hundred organic compounds were screened, and two disubstituted furans (A1 and A5) with new structure showed inhibition of VEGF receptor 1 kinase. Compound A1 inhibited only phosphorylation of substrate, while compound A5 inhibited both autophosphorylation and substrate phosphorylation. Both inhibitors affected phosphorylation in the transformed cells. CONCLUSION: The recombinant receptor kinase based assay is simple and effective in identifying kinase inhibitors. PMID- 11866871 TI - Effects of simvastatin on activities of endogenous antioxidant enzymes and angiotensin-converting enzyme in rat myocardium with pressure-overload cardiac hypertrophy. AB - AIM: To investigate effects of simvastatin (Sim) on the activities of antioxidant enzymes and angiotensin-converting enzyme in rat myocardium with pressure overload cardiac hypertrophy. METHODS: Left ventricular hypertrophy (LVH) was induced by partly constricting rat abdominal aorta between the left and right renal artery. Rats were given ig Sim 1.8 and 3.6 mg . kg-1 . d-1 for 8 weeks following 6 weeks aortic constriction. Activities of antioxidant enzymes and angiotensin-converting enzyme, and lipid peroxidation of left ventricular (LV) tissue were determined. RESULTS: Contents of angiotensin II and thiobarbituric acid reactive substances (TBARS), and activity of ACE in LVH group (n = 8) were increased by 163 %, 90 %, and 130 %, respectively (P < 0.01)compared with sham operated group (n = 7), and were decreased by 30 %, 37 %, and 51 %, respectively (P < 0.01) in high dosage Sim treatment group (n = 9) compared with LVH group. Activities of catalase and glutathione peroxidase of LV tissue in LVH group were decreased by 29 % and 23 % (P < 0.01) compared with sham-operated group, and were increased by 32 % and 22 % (P < 0.01) in high dosage Sim treatment group compared with LVH group. Activity of Cu, Zn-superoxide dismutase (SOD) of LV tissue was increased by 33 % in LVH group compared with sham-operated group. Sim treatment did not significantly affect activity of Cu,Zn-SOD. CONCLUSION: Alteration of redox status in myocardium is associated with cardiac hypertrophy and inhibitory effects of Sim on cardiac hypertrophy in rats model might be linked to its antioxidant effects. PMID- 11866872 TI - Inhibitory effects of praeruptorin C on cattle aortic smooth muscle cell proliferation. AB - AIM: To study the effects of praeruptorin C (pra-C) on proliferation of cattle aortic smooth muscle cells (SMC). METHODS: The DNA synthesis of SMC was measured using the incorporation of [3H]thymidine([3H]TdR). Cell cycle phase was evaluated by flow cytometry and cytotoxicity was evaluated by measuring lactic dehydrogenase (LDH) activity. RESULTS: Whether or not treated with angiotensin II (Ang-II), SMC proliferation was suppressed by pra-C in a concentration-dependent manner at range from 0.001 micromol/L to 10 micromol/L. The inhibitory effects appeared to be related to G1-S block in cell cycle traverse while the LDH activities did not change dramatically. CONCLUSION: Pra-C can completely inhibit SMC proliferation induced by Ang II and partly inhibit the growth of SMC- induced by bovine serum, which is important in prevention and treatment of vascular hyperplastic disease. PMID- 11866873 TI - Comparison of norepinephrine-dobutamine to dopamine alone for splanchnic perfusion in sheep with septic shock. AB - AIM: To compare the effect of norepinephrine-dobutamine with dopamine alone on splanchnic perfusion in sheep with septic shock. METHODS: Twenty sheep with septic shock induced by lipopolysaccharides were divided into two groups. When systolic pressure decreased by 5.3 kPa, basic values of hemodynamic parameters and intestinal intramucosal pH (pHi) were recorded. Each group was randomized to receive an intravenous infusion of norepinephrine-dobutamine or dopamine, and titrated to obtain mean arterial pressure (MAP) > 12 kPa with an optimal cardiac preload. Hemodynamic parameters and mucosal pHi were repeated at 1, 2, 3, and 4 h after basic measurement. RESULTS: After norepinephrine-dobutamine or dopamine infusion, MAP, cardiac output, and oxygen delivery increased in all animals compared with basic values in both groups (P < 0.05). Compared with baseline values, lactate concentrations decreased at 3 h and 4 h [from (4 +/- 2) mmol/L to (2 +/- 1) mmol/L] in the norepinephrine-dobutamine group (P < 0.05 ). Arterial lactate concentrations had no change in dopamine group, but arterial pH decreased from 7.40 +/- 0.05 to 7.26 +/- 0.06 at 1 h (P < 0.05 ). No difference in pHi was found in dopamine group, but in the norepinephrine-dobutamine group, compared with baseline, pHi increased from 7.19 +/- 0.04 to 7.36 +/- 0.07 at 3 h (P < 0.05). CONCLUSION: Both norepinephrine-dobutamine and dopamine alone could improve systemic hemodynamics in sheep with septic shock, but norepinephrine dobutamine was better than dopamine on splanchnic perfusion. PMID- 11866874 TI - Inhibitory effect and mechanism of action of sanggenon C on human polymorphonuclear leukocyte adhesion to human synovial cells. AB - AIM: To examine the effect of sanggenon C on human polymorphonuclear leukocyte (PMN) adhesion to human synovial cell(HSC), and explore its mechanism. METHODS: Adhesion of PMN to HSC was measured by MTT colorimetry. Cell-ELISA and RT-PCR methods were used to examine the expression of adhesion molecules ICAM-1 and VCAM 1. Activation of nuclear factor-kappa B(NF-kappaB) was measured by electrophoretic mobility shift assays(EMSA) method. RESULTS: Sanggenon C effectively inhibited TNF-alpha (50 kU/L for 12 h) and IL-1beta (50 kU/L for 12 h) induced adhesion of PMN to HSC (IC50 27.29 nmol/L and 54.45 nmol/L, respectively) in a concentration-dependent manner. Adhesion molecule VCAM-1 surface protein and mRNA expression induced by TNF-alpha 50 kU/L were significantly inhibited by sanggenon C, nevertheless, for ICAM-1 only surface protein expression being inhibited. The activation of NF-kappaB was also extensively inhibited by sanggenon C. CONCLUSION: Sanggenon C inhibited TNF-alpha -stimulated PMN-HSC adhesion and expression of VCAM-1 by suppressing the activation of NF-kappaB. PMID- 11866875 TI - Expression and purification of catalytic domain of human macrophage elastase for high throughput inhibitor screening. AB - AIM: To obtain a catalytically active human macrophage elastase catalytic domain (hMECD) and to establish an efficient high-throughput method for screening macrophage elastase inhibitors. METHODS: Catalytic domain of human macrophage elastase was expressed in E coli and characterized to establish a high-throughput screening assay using a colorimetric method. A set of 8560 pure compounds and mixtures were screened. RESULTS: We have constructed an efficient E coli system for this human protein expression, and the recombinant hMECD protein was purified to homogeneity using anion-exchange chromatography after in vitro refolding from inclusion bodies. The yield of active hMECD protein was 23 mg from one liter of E coli culture after purification. Calcium and zinc ions were required both in refolding and enzymatic activity, but high concentration of zinc inhibited the refolding and activity. The hMECD cleaved several synthetic substrates including a chromogenic thiopeptolide and fluorogenic peptides with optimal activity around pH 8.0. Screening of 8560 compounds and mixtures led to identify 27 pure compounds and 14 natural products with inhibitory activity higher than 80 % at 20 mg/L. CONCLUSION: An efficient expression and purification method for hMECD protein has been established, and the assay is effective, reliable, and fast in identifying the recombinant protein inhibitors. PMID- 11866877 TI - VCAM-1 expression, eosinophil infiltration, and pharmacological modulation in rat allergic airway inflammation. AB - AIM: To determine vascular cell adhesion molecule-1 (VCAM-1) expression and eosinophil infiltration in the lungs of the rats with allergic airway inflammation, and their possible modulation by dexamethasone and neurokinin-1 receptor antagonist SR140333. METHODS: Sensitized rats were challenged with inhalation of 1 % ovalbumin aerosol. Protein expression of VCAM-1 in the lungs and eosinophil infiltration around bronchi in different groups were determined 24 h after challenge. SR140333 (0.01 and 0.10 mg/kg) and dexamethasone (0.50 mg/kg) were injected ip, twice a day, for 3 d before challenge. RESULTS: The expression of VCAM-1 was increased in the sensitized rat lungs as compared with the non sensitized rats (P<0.05). The increment was inhibited by pretreatment with dexamethasone (P<0.05), but not with SR140333 (P>0.05). On the other hand, antigen challenge in sensitized rats evoked eosinophil infiltration (P<0.05) but no further increase in VCAM-1 expression (P>0.05). Furthermore, SR140333 inhibited eosinophil infiltration (P<0.05) but had no effect on VCAM-1 expression (P>0.05), whereas dexamethasone inhibited both responses (P<0.05). CONCLUSION: The expression of VCAM-1 increases during antigen sensitization in rat lungs, and dexamethasone and SR140333 may inhibit allergic airway inflammation in different mechanisms. PMID- 11866876 TI - Acetamide-45 inhibits histamine-and methacholine-induced contraction of isolated guinea pig trachea. AB - AIM: To investigate the effects of the new antiallergic agent N-(pyridin-4-yl) (indol-3-yl) acetamide-45 (acetamide-45) on histamine- and methacholine-induced contraction of the isolated guinea pig trachea. METHODS: Cumulative histamine or methacholine concentration-contraction studies were carried out in the absence or presence of acetamide-45. Changes in isometric force were measured by force transducers and recorded on a multi-channel polygraph recorder. RESULTS: Acetamide-45 (1 - 30 micromol/L) concentration-dependently inhibited histamine- or methacholine-induced contractile response of isolated guinea pig trachea. At concentrations of 3, 10, 30 mumol/L, acetamide-45 significantly decreased maximum contractile response to histamine by 21 % - 51 %, and EC50 values (95 % confidence limits) were 31.1 (24.4 - 39.8), 34.7(26.8 - 45.0), and 134.4 (82.2 - 220.0) micromol/L, respectively. Similarly, acetamide-45 also inhibited the contraction induced by methacholine in isolated guinea pig trachea. CONCLUSION: Acetamide-45 inhibited histamine- or methacholine-induced contraction of isolated guinea pig trachea, and these effects might be non-specific for either histamine receptor or cholinoceptor. PMID- 11866878 TI - Structural characterization and biological activities of SC4, an acidic polysaccharide from Salvia chinensis. AB - AIM: To study the chemical characterization and some biological activities of an acidic polysaccharide, named SC4 from Salvia chinensis. METHODS: Chemical and spectral methods were employed to identify the structural characterization of SC4. Effect of SC4 on the proliferation of T- and B-lymphocytes both in vivo and in vitro was measured at various concentrations, and the protective effects of SC4 on PC12 cells against H2O2-induced injury was observed. RESULTS: SC4 was a highly branched polysaccharide with mean molecular weight of 4.5 10(5), composed of Rha, Xyl, Ser, Gal, and GalA in the molar ratio of 1.0 7.0 5.3 1.2 4.2. Methylation analysis and NMR identified the linkages of the residues of SC4. SC4 increased the B-lymphocyte proliferation and spleen weight remarkably while no obvious changes were examined in T-lymphocyte proliferation, thymus weight, and the level of IgG and C3 in blood of mice. Moreover, SC4 enhanced the PC12 cells viability after H2O2 treatment. CONCLUSION: SC4, the acidic polysaccharide with complicated structure, was a B-lymphocyte stimulator and protected PC12 cells at the concentration of 20 mg/L against H2O2-induced injury. PMID- 11866879 TI - Synergism between heparin and adriamycin on cell proliferation and apoptosis in human nasopharyngeal carcinoma CNE2 cells. AB - AIM: To measure the effect of addition of heparin to adriamycin (ADM) on cell proliferation and apoptosis in CNE2 cells and investigate the possible molecular mechanisms of heparin and ADM interactions. METHODS: Cell viability and cell cycle were determined by MTT assay and flow cytometry. Apoptosis was evaluated by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) and agarose gel electrophoresis. The expression of Bax, Bcl-2, p53, and p21 was examined by Western blot. RESULTS: ADM (5 mg/L) alone inhibited the growth of CNE2 cells, which was magnified when heparin was added. ADM elicited typical apoptotic morphologic changes. Compared with ADM or heparin alone, ADM plus heparin obviously enhanced the number of TUNEL positive cells from 12.6 % 1.1 % to 65.7 % 1.3 %, and the DNA ladder was more clearly observed. After exposure to different concentrations of heparin (with or without ADM) for 24 h, CNE2 cells were accumulated in G0/G1 phase. There was a decrease in the number of cells in S phase by the combined heparin and ADM treatment compared to heparin or ADM alone. The ratio of Bax/Bcl-2 was elevated, and p53 and p21 mRNA were over-expression. CONCLUSION: Heparin and ADM appear to interact in a synergistic manner, which may be related to the over-expression of p53 and p21 mRNA and the elevated ratio of Bax/Bcl-2 mRNA. PMID- 11866880 TI - Isolation of acetylcholinesterase G4 and G1 molecular isoforms from rat cortex. AB - AIM: To isolate the G4 and G1 molecular forms of acetylcholinesterase in rat cortex for interpreting the therapeutical effect of the AChE inhibitor drugs. METHODS: Size exclusion chromatography and ultracentrifugation were used to isolate and ascertain the G4 and G1 molecular forms of AChE. RESULTS: After size exclusion chromatography, two AChE active peaks were gotten. The putative molecular weights of the two highest AChE active fractions were around Mr 239 000 and Mr 68 000 respectively, which represented G4 and G1 AChE isoforms. The sedimentation coefficients of the Mr 239 000 and Mr 68 000 fractions were 10 S and 4 S, which also assigned to G4 and G1 AChE isoforms. CONCLUSION: The G4 and G1 isoforms of AChE could be gotten by size exclusion chromatography method. PMID- 11866882 TI - Alzheimer-like tau phosphorylation induced by wortmannin in vivo and its attenuation by melatonin. AB - AIM: To investigate the in vivo induction of Alzheimer-like tau phosphorylation by wortmannin and the attenuation by melatonin. METHODS: Stereotaxic technique was used for administrating wortmannin or melatonin into rat lateral ventricle. The phosphorylation of tau was analyzed by immunocytochemistry and Western blot, and ultrastructural alteration in neuronal processes was detected by electron microscopy. RESULTS: Level of phosphorylated tau at paired helical filament-1 (PHF-1) epitope was elevated at 1 h, peaked at 6 h, and then decreased at 12 h and 24 h after injection of wortmannin 10 mumol/L. The increased tau phosphorylation at particular epitope determined at 6 h was arrested by preinjection of melatonin 10 mumol/L or 100 mumol/L for 12 h. The wortmannin induced hyperphosphorylation of tau was mainly detected in pyramidal neuron of hippocampus, and swelled axons and depredated myelin sheaths were also seen in the same region of the brain. CONCLUSION: Wortmannin induced in vivo Alzheimer like hyperphosphorylation of tau at Ser396/404, and melatonin inhibited partially the pathological processes. PMID- 11866881 TI - Effect of mifepristone on uterine receptivity in guinea pigs. AB - AIM: To determine the effect of a low-dose mifepristone regimen on endometrial receptivity in guinea pigs. METHODS: Histologic evaluation for endometrial staging and immunoreactivity of cytokines [leukemia inhibitory factor (LIF) and interleukin-6 (IL-6)] were preformed on fixed endometrial tissue, and relative abundance of endometrial estrogen (E), and progesterone (P) receptor mRNA were evaluated with semiquantitative reverse transcriptase polymerase chain reaction (RT-PCR) in which cyclophilin mRNA, a housekeeping gene product, was co-amplified as the reference standard. RESULTS: Mifepristone at 0.03 mg.kg-1 . d-1 induced a delay in endometrial maturation. The staining intensity of LIF and IL-6 decreased in treated guinea pigs. E and P receptor mRNA increased significantly and uterine stage delayed in treated guinea pigs. CONCLUSION: Mifepristone at 0.03 mg.kg-1 . d-1 delayed development of endometrium. This study provides further evidence that endometrial maturation can be altered without affecting ovarian ovulatory function. PMID- 11866883 TI - Nonassociation of aryl hydrocarbon receptor genotypes with susceptibility to bladder cancer in Shanghai population. AB - AIM: To assess two polymorphic forms of aryl hydrocarbon receptor (AHR) gene, G1721A (R554K), and G1768A (V570I) in Chinese population and to explore the possible association of human AHR gene polymorphism with elevated incidence of bladder cancer among Chinese Han subjects in east of China. METHODS: An allele specific PCR-based procedure for AHR gene polymorphism genotyping was developed by this work. Genotyping on three groups of subjects in Shanghai area had been performed: a bladder cancer group with the occupational exposure to benzidine, a non-occupational bladder cancer patient group whose members lack an obvious aromatic amine exposure record, and a normal population in the same city as controls. RESULTS: A significant difference (P < 0.01) in frequency distribution at locus G1721A between normal population in Shanghai and a Caucasian population reported by other authors was observed. No mutant allele(A1768) at locus G1768A had ever been detected in our study. The observed frequencies were similar between both genders in the normal population (P = 0.54), and there were no significant difference confirmed between the case group and the control group. CONCLUSION: The locus G1768 of human AHR gene seems to be monomorphic among Chinese in this area. The significant distribution difference at locus G1721A of human AHR gene between Chinese Han and Caucasian was confirmed. This study did not support the association of AHR G1721A polymorphism with higher risk to bladder cancer among the residents in this area, either in a group of occupationally benzidine-exposed individuals or among the persons who never have an obvious aromatic amine exposure record. PMID- 11866884 TI - [Relationship of bcl-2 gene expression with cell proliferation and apoptosis in human gliomas]. AB - OBJECTIVE: To investigate the relationship of bcl-2 gene expression level in human gliomas with the malignant degree, cell proliferative activity and apoptosis of the tumors. METHODS: The expression of bcl-2 mRNA, bcl-2 protein and proliferating cell antigen, and the apoptosis in sixty-nine human glioma specimens with different malignant grades were studied using in situ hybridization, in situ cell death detection (TUNEL method) and immunohistochemistry. RESULTS: Of the 69 gliomas, 64 (92.8%) and 60 (87.0%) expressed bcl-2 mRNA and bcl-2 protein, respectively. The expression levels of bcl-2 mRNA and bcl-2 protein were correlated positively with each other (r(s) = 0.999, P < 0.01). The expression levels of bcl-2 mRNA and bcl-2 protein were both higher in WHO grade IV gliomas than in grade III gliomas, and the expression was lowest in grade I-II gliomas (P < 0.02 approximately 0.001). With the increase in the expression of bcl-2 protein, the cell proliferating activity increased and apoptosis decreased in the tumor cells. There was significant difference of cell proliferation and apoptosis between + + + group and + + group of bcl-2 protein expression (P < 0.05) as well as between both the former groups, and the negative and + group (P < 0.01) respectively. CONCLUSIONS: Over-expression of bcl-2 gene inhibits apoptosis of glioma cells, and the inhibitory intensity increases with the ascending of bcl-2 gene expression level in glioma cells. Both the decrease in apoptosis caused by bcl-2 gene over-expression and the excessive cell proliferation promoted by other gene abnormalities may result in unlimited cell accumulation, which may play an important role in the development and malignant progression of gliomas. PMID- 11866885 TI - [Expression of telomerase genes in human tumors]. AB - OBJECTIVE: To investigate the correlation between the expression of telomerase genes and malignant phenotypes of human tumors and to compare the expression of telomerase genes and its activity reported in order to evaluate the role of detection of telomerase genes in tumor diagnosis. METHODS: With in situ hybridization, the expression and distribution of telomerase hTR and hTRT genes were observed in 78 cases of human cancer tissues, 20 cases of precancerous lesions and 28 of benign lesions. The results were statistically analyzed. RESULTS: hTR and hTRT were detected in 85% (66/78) and 82% (64/78) of the primary cancers. While in the adjacent tissues, the positive rates were only 3% (2/78) and 5% (4/78). In 20 precancerous cases, the positive rate for hTR and hTRT were 20% (4/20) and 15% (3/20) and the positive cases in 28 benign lesions were 0 and 1/28 (4%), respectively. The positive detection of hTR and hTRT expression in cancer group were significantly different from those in the adjacent tissues, precancerous cases and benign lesion group (P < 0.01). In analysing main types of the human cancers, the positive frequency of hTR and hTRT were 94% (15/16) and 88% (14/16) for the breast cancer, 85% (17/20) and 90% (18/20) for the colon cancer, 80% (8/10) and 80% (8/10) for the gallbladder cancer, 75% (6/8) and 75% (6/8) for the lung cancer, 75% (6/8) and 75% (6/8) for the stomach cancer, and 83% (5/6) and 83% (5/6) for the esophagus cancer, respectively. The expression level of telomerase hTR and hTRT correlated well with the malignancy and metastatic potentials in breast cancer, colon cancer and bladder cancer. Besides, the expressions of hTR and hTRT were noticed to be also highly correlated (P < 0.01). CONCLUSIONS: The expression of telomerase genes correlates with tumor malignant phenotypes, and may reflect the progression of tumors, and the detection of telomerase gene expression may be useful in a retrospective cancer research. It is worthwhile for further study to clarify whether screening of telomerase gene expression is able to become a new index for tumor diagnosis and prognosis. PMID- 11866886 TI - [p16, p53 and c-erbB-2 gene expression in bladder carcinoma]. AB - OBJECTIVE: To investigate the expression of p16, p53 and c-erbB-2 proteins in bladder carcinoma and to analyze their correlation with tumor grade, clinic stage and metastasis. METHODS: Immunohistochemical method was used to detect the p16, p53 and c-erbB-2 gene expressions in 75 cases of bladder carcinoma. RESULTS: Positive rates of p16, p53 and c-erbB-2 in 75 cases of bladder cancer were 41.3%, 44.0% and 40.0%, respectively. p16 and c-erbB-2 gene expressions were related to the tumor grade and clinic stage (P < 0.05); p53 and c-erbB-2 gene expressions were related to the clinic stage and metastasis (P < 0.01). Oncogenes and/or antioncogenes altered expressions were found in 77.3% cases of bladder carcinoma, and oncogenes and/or antioncogenes with multiple altered expressions were found in 53.3% cases. CONCLUSION: The analysis of multiple oncogene and/or antioncogene is more valuable than that of single oncogene and/or antioncogene. The altered expression of oncogene c-erbB-2 and antioncogenes p16 and p53 might play a promotive role in the tumorigenesis and development of bladder tumors. PMID- 11866887 TI - [Analysis of TCR-gamma gene rearrangement in single H/R-S cells of Hodgkin disease]. AB - OBJECTIVE: To detect the rearrangement of T cell receptor gamma chain gene in single H/R-S cell as an indication of early T lymphocyte differentiation. METHODS: CD30 positive H/R-S cells were isolated with micromanipulator from the frozen sections and analyzed by single-cell PCR. TCR-gamma chain gene rearrangement was detected through semi-nested PCR. In the first round of PCR, a pair of TCR-gamma primers which amplify the segments between Vgamma and Jgamma region (including the N region) of TCR-gamma gene mixing with a pair of beta globin primers were used. In the second round, only the products positive beta globin were amplified with the primers for TCR-gamma gene. RESULTS: A 268 bp beta globin gene specific product was obtained in 24 out of 56 H/R-S cells from 6 cases of Hodgkin's disease. After the first round of PCR, no rearrangement of the TCR-gamma gene was noticed at a single cell level. CONCLUSION: H/R-S cells from the selected cases were not derived from the T-cell lineage but possibly from the B-cell lineage. PMID- 11866888 TI - [Effects of platelet-derived growth factor on the proliferation of hepatic stellate cells and their expressions of genes of collagens and platelet-derived growth factor]. AB - OBJECTIVE: To investigate the effects of PDGF on the proliferation of rat HSC, its gene expression of collagens and the autocrine excretion of PDGF in vitro. METHODS: (3)H-TdR incorporation was used to estimate the DNA synthesis elicited by PDGF, TGF-beta1 and EGF in HSC cultured in an absolutely serum-free medium. Northern blot was employed to detect the levels of mRNA expressions of type I and III procollagens and PDGF induced by PDGF added in the same medium for HSC. RESULTS: Both PDGF and EGF enabled to stimulate the proliferation of HSC in a dose-dependent manner, and the effect of the former was stronger. The proliferation was dose-dependently increased in PDGF and EGF with a dosage between 0.5 and 10 ng/ml, and the synthesis of DNA became saturated when the dosage was over 10 ng/ml. TGF-beta1 gave no effects on the proliferation of HSC under current experimental condition. Furthermore, PDGF was able to enhance the mRNA expression for type I and III procollagens and PDGF itself. The promoting effect for the expressions of type I and III procollagen mRNAs by PDGF was bi phasic between 6 to 48 hours of the cultivation. For type I procollagen, mRNA expression began to increase 12 hours after PDGF stimulation, and the increase became prominent at 48 hours. For type III procollagen, an increase of mRNA expression was seen before the stimulation of PDGF, and the expression level was comparatively increased 48 hours later. The mRNA expression of PDGF was increased by 2-3 times 6 hours after PDGF stimulation, and back to the original level 24 hours later. CONCLUSIONS: PDGF enabled to promote the proliferation of HSC and expression of collagens in HSC. The bi-phasic effect on the expression of collagens may be interrelated with the autocrine excretion of PDGF. PMID- 11866889 TI - [The effect of nordihydroguaiaretic acid on endothelial cell migration induced by glioma cells]. AB - OBJECTIVE: To investigate the effect of nordihydroguaiaretic acid (NDGA) on the migration of endothelial cells (ECs) induced by glioma cells (GCs) in vitro. METHODS: Cells of human umbilical vein endothelial cell line ECV-304 and human malignant glioma cell line SHG-44 were co-cultured in the Falcon Cell Culture Insert system, and the effect of NDGA on the migration of ECs induced by GCs was investigated. The expression of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) was examined with immunohistochemistry. RESULTS: The expressions of VEGF and bFGF in SHG-44 cells were reduced after treatment with 100 micromol/L NDGA for 1 to 3 days. 100 micromol/L NDGA significantly inhibited not only the chemotactic migration of ECs induced by either glioma cells or glioma-conditioned media, but also the random motility of ECs (P < 0.01). NDGA at a concentration between 50 micromol/L and 200 micromol/L inhibited the chemotactic migration of ECs in a dose-dependent manner (the inhibition ratios were 12.0%, 37.4%, 86.7% and 82.2%). CONCLUSION: The results confirm the inhibitory effects of NDGA on the migration and angiogenic factor expression of ECs, which suggests that NDGA may suppress angiogenesis of glioma. PMID- 11866891 TI - [Malignant transformation of mouse embryonic fibroblast induced by mitochondrial DNA fragments]. AB - OBJECTIVE: To investigate the malignant transforming effect and mechanism of mitochondrial DNA (mtDNA) fragments. METHODS: Tumorigenicity of mtDNA-transformed mouse embryonic fibroblast (NIH3T3) in nude mice was studied using transgenic techniques. Transformed tumors were detected by pathological examination and hybridization signals of mtDNA probe were analyzed by fluorescence in situ hybridization (FISH) techniques. RESULTS: Hybridization signals were observed on the nuclei of 18% - 20% NIH 3T3 cells 1 week after mtDNA fragments transforming. Tumor from mtDNA-transformed NIH 3T3 cells was developed in all 8 nude mice (8/8) respectively 2 weeks after the transformation. The pathological characteristics of the tumors developed were similar to that of fibrosarcoma. CONCLUSIONS: Auto integration of mtDNA fragments into nuclear genome is a new factor involved in carcinogenesis. PMID- 11866890 TI - [Different susceptibility of cell lines to thymidine kinase/ganciclovir-mediated killing effect and the way of cell death]. AB - OBJECTIVE: To investigate the mechanism of variability of herpes simplex virus (HSV) thymidine kinase (TK)/ganciclovir (GCV)-mediated suicide effect obtained in three different cell lines and the ways in which this system-mediated cell killing occurs. METHODS: Recombinant retroviral vector expressing HSV-TK was transduced into three cell lines known with different growth rate (PC-2, PC-7 and LLC-PK1). The doubling time of the transduced and parental cells was calculated. MTT method was used to detect the concentration of GCV at which cell growth was inhibited by 50% (IC(50) value). Cell cycle was analyzed by flow cytometry. Necrosis or apoptosis of cultured cells and/or xenografts was observed under light and electron microscope, and by in situ apoptosis detection. RESULTS: The doubling time of the three parental cell lines was (31.2 +/- 0.1) h, (48.3 +/- 0.1) h, and (53.9 +/- 0.1) h, separately. The IC(50) values of their HSV-TK transduced cell lines to GCV were (0.73 +/- 0.12) micromol/L, (0.93 +/- 0.16) micromol/L and (1.22 +/- 0.06) micromol/L, respectively and the IC(50) value was correlated with the cell doubling time. Flow cytometry revealed S arrest. The majority of cells under treatment of GCV displayed swelling and collapse, but very few cells showed apoptosis. Large areas of necrosis were observed in the xenografts. CONCLUSIONS: The cells with high growth rate are more susceptible to HSV-TK/GCV-mediated killing effect. Necrosis is the main way in which cells of the three HSV-TK-transduced cells lines die. PMID- 11866892 TI - [Changes of surfactant A and B in alveolar type II cells in hamster with elastase induced emphysema]. AB - OBJECTIVE: To observe the changes of surfactant A and B (SP-A and SP-B) in alveolar type II cells of lungs in hamsters with elastase-induced emphysema. METHODS: After intratracheal instilling of elastase, hamsters were killed at the 30th, 60th and 90th day of the experiment respectively. Slides of lung tissue were examined under light microscope and measured by calculating the mean linear intercepts. Then the slides were stained under immunohistochemistry procedures and measured by using a morphometric analysis system. Meanwhile, slides were also checked by electron microscopy. RESULTS: In comparison with the normal group, MLI of the elastase group was significantly increased (P < 0.01). Immunohistochemistry showed that the percentage of both SP-A and SP-B positive cells was significantly decreased 30 days after the intratracheal instilling of elastase (from 48.0% +/- 3.0% to 9.5% +/- 4.0% and from 28.0% +/- 4.0% to 13.3% +/- 4.1%, respectively; P < 0.01), however, a time-dependent recovery 90 days after instilling was seen. Electron microscopy showed a significantly decrease of the number of lamellae. CONCLUSION: The decrease of SP-A and SP-B in alveolar type II cells seems to play an important role in the development of emphysema. PMID- 11866893 TI - [Effects of IL-1beta, TNF-alpha and lipopolysaccharide on the expression of MCP-1 in human umbilical vein endothelial cells]. AB - OBJECTIVE: To investigate whether the proinflammatory cytokines IL-1beta, TNF alpha and lipopolysaccharide (LPS) enable to induce the expression of MCP-1 mRNA and protein in human umbilical vein endothelial cells (HUVECs). METHODS: After a four-hour exposure to 2 ng/ml IL-1beta, 20 ng/ml TNF-alpha or 100 ng/ml LPS, total RNA of HUVECs was extracted by single-step method. The expression of MCP-1 mRNA in HUVECs was examined by dot blot analysis using a probe of gamma-(32)P-end labelled 35 mer oligonucleotide. Meanwhile, MCP-1 protein in the cytoplasm was detected by SABC immunostaining. RESULTS: Dot blot analysis showed that cultured HUVECs were able to express MCP-1 mRNA at a low level. Exposure to IL-1beta, TNF alpha and LPS resulted in a 7.8-fold, 2.6-fold and 1.2-fold induction of MCP-1 mRNA expression in HUVECs, respectively. The cells on the coverglass in all groups revealed MCP-1 immunoreactivity. Densitometry scans showed that the mean absorbance (A) values of the cells in LPS, TNF-alpha and IL-1beta groups were 0.078 +/- 0.113, 0.102 +/- 0.005 and 0.117 +/- 0.010, respectively; whereas the absorbance values of the control group was 0.051 +/- 0.004. There were significant differences between all the experimental groups and the control group (F = 193.25, P < 0.01). CONCLUSIONS: IL-1beta and TNF-alpha induce a strong expression of MCP-1 mRNA and protein in HUVECs. Both cytokines may be involved in the atherogenesis by inducing the liberation of MCP-1 by endothelial cells and increasing the recruitment of monocytes into the subendothelial space. PMID- 11866894 TI - [Effect of sodium fluoride on bone turnover and bone loss of vertebrae and tibia in ovariectomized rats]. AB - OBJECTIVE: To investigate the effect of sodium fluoride on bone turnover and bone loss of vertebra and tibia of ovariectomized rats. METHODS: 34 female Wistar rats, 16 weeks old, were divided into 3 groups: sham-ovariectomy (CON), bilateral ovariectomy (OVX) and both OVX and NaF-treatment groups (OVX + NaF, 1.0 mg/kg BW). All of the rats were sacrificed 12 weeks after the treatment. Serum was collected for the analysis of estradiol, calcium, phosphorum and alkaline phosphotase. Temur was used for the analysis of calcium content. Tibiae and the first lumbar vertebral bodies were decalcified for histomorphometry. RESULTS: After treatment with NaF, bone resorption was not inhibited, but bone formation was significantly increased. The trabecular bone volume of vertebrae (18.73% +/- 3.04%) and tibiae (14.30% +/- 5.87%) was significantly increased in comparing with of OVX rats (15.07% +/- 2.45% and 8.81% +/- 2.60%, P < 0.05), but it was markedly lower than that of CON rats (22.44% +/- 1.82% and 24.87% +/- 4.02%, P < 0.05 and P < 0.01). CONCLUSIONS: NaF could not decrease bone turnover and it, has only a partial protective effect against bone loss of either vertebra or tibia. PMID- 11866895 TI - [Loss and inactivation of PTEN/MMAC1/TEP1 gene in lung cancer]. AB - OBJECTIVE: The recently identified PTEN/MMAC1/TEP1 gene is the first tumor suppressor encoding a phosphatase, which mutated in multiple tumors. Our research aims to detect loss of heterozygosity and inactivation of PTEN in lung cancer. METHODS: 24 lung cancer fresh samples paired with normal tissue, 18 archival paraffin-embedded sections of small cell lung cancer (SCLC) were included. For 24 fresh samples, PCR-LOH was used to detect Loss of heterozygosity (LOH) in or near PTEN loci. In situ hybridization, Western blot and or immunohistochemistry were proceeded for mRNA and protein expression of PTEN in lung cancer samples. Southern, Northern and Western blot were done to exam PTEN abnormality in one NSCLC and three SCLC cell lines. RESULTS: Among these 24 fresh samples PTEN LOH was detected in 11 out of 24 cases (45.8%) and 6 cases had no PTEN mRNA and protein expression. Results in situ hybridization were in conformity with that of immunohistochemistry/Western blot in our research. No protein expression was detected in 8/18 (44%) archival paraffin cases of SCLC. Southern, Northern and Western blot confirmed homozygous deletion in one SCLC cell line. CONCLUSION: Lung cancers had LOH and inactivation of PTEN. PMID- 11866896 TI - [Telomerase activity in human non-small cell lung cancer (NSCLC)]. AB - OBJECTIVE: To investigate the activity of telomerase in human non-small cell lung cancer (NSCLC) and the correlation of telomerase activity with the cancer development. METHODS: Totally 48 cases of NSCLC tissues were studied accompanied with 7 cases of normal lung tissues and 12 samples of lung tissues adjacent to the tumor. The telomerase activity was detected using Telomerase PCR ELISA kit. Statistics study included chi(2)-test and the exact probabilities. The significance level chosen was P value less than 0.05 (P < 0.05), and all tests were two-sided. RESULTS: 75% (36/48) of NSCLC tissues possessed the expression of telomerase activity and only 8.33% (1/12) of tissues adjacent to tumor and none of the normal lung tissue enabled to express telomerase activity. The statistical difference was significant (P < 0.05). No significant difference was found between different histological types of NSCLC, however, the frequency of telomerase expression differed significantly between tumors of well/mordeately differentiated and poorly differentiated; stage I and stage II/III. CONCLUSION: Telomerase activity might be a good marker in estimating the occurrence, development and diagnosis of NSCLC. PMID- 11866897 TI - [Study on spontaneous apoptosis of human non-small cell lung cancer tissues]. AB - OBJECTIVE: To observe the spontaneous apoptosis in human non- small cell lung cancer (NSCLC) tissues, and the impact of PCNA and microvessel density on it. METHODS: TDT in situ labeling and immunohistochemistry were used to display apoptotic cells, PCNA and microvessels. RESULTS: The total forty-six cases of NSCLC were observed. Apoptosis index (AI) was 2.8% +/- 2.3% on average, and higher in squamous carcinomas (3.4% +/- 1.7%) than in adenocarcinomas (1.2% +/- 2.0%). AI in the poorly differentiated carcinomas (3.9% +/- 2.7%) was significantly higher than in the middle- and well-differentiated carcinomas (2.8% +/- 2.0%, 0.5% +/- 0.2% respectively). AI was higher in NSCLC with PCNA-positive cell rate >or= 25% (3.4% +/- 2.2%) than with PCAN-positive cell rate < 25% (1.9% +/- 2.2%). Microvessel density was higher in NSCLC with AI < 2% ( 74.4 +/- 29.6 counts/200 field) than with AI >or= 2% (42.0 +/- 12.8 counts/200 field). Apoptotic cells were frequently seen in the regions with less microvessels and distal to microvessels. CONCLUSIONS: Spontaneous apoptotic cells mainly occur in the tissues of squamous carcinomas and poorly-differentiated carcinomas and NSCLC with higher PCNA-positive cell rates. Hypoxia could induce apoptosis of lung cancer cells. PMID- 11866898 TI - [Methylation status of p16 gene in colorectal carcinoma and normal colonic mucosa]. AB - OBJECTIVE: To investigate the methylation status and its significance of 5' CpG island of p16 gene in colorectal carcinoma and normal colonic mucosa. METHODS: PCR-based methylation assay was applied to examine a total of 60 samples of colorectal carcinoma and paired 60 samples of the normal colonic mucosa for p16 gene methylation in Hpa II, Sac II, Sma I sites. RESULTS: Methylation was identified in 5' CpG island of p16 gene in both colorectal carcinoma and normal colonic mucosa, but no significant difference obtained between normal mucosa and carcinoma. The methylation status in 5' CpG island of p16 gene in colorectal carcinoma was not related to the tumor size, type, and metastases of the lymph nodes. Nevertheless, there was a significant difference between the methylation status of Hpa II, Sac II and Sma I sites. CONCLUSIONS: The methylation in 5' CpG island of p16 gene in colonic mucosa is different in various restriction enzyme sites, and the methylation process of p16 gene in both normal colonic mucosa and colorectal carcinomas is complicated. Anyhow, there is a tendency that the increase of methylation in p16 gene is age dependent. PMID- 11866899 TI - [The expression and clinical significance of p80 protein in anaplastic large cell lymphoma]. AB - OBJECTIVE: To investigate the relationship of expression of NPM-ALK cohesive gene protein with the subtypes and prognosis of anaplastic large cell lymphoma (ALCL). METHODS: Nineteen cases of ALCL with CD30 positivity were studied. Immunohistochemical staining was performed in paraffin sections by using polyclonal antibody anti-p80 and other monoclonal antibodies. The clinical data of these patients were analyzed and all patients were followed-up. RESULTS: The neoplastic cells in 9 out of 19 ALCL expressed p80 protein. Histopathologically, all 9 cases with p80 expression were common type and small cell type. None of these 9 cases were Hodgkin like type (P < 0.05). Immunologically, all the 9 positive cases were T and null cell phenotype and none of them was B cell or other phenotype (P < 0.05). Clinically, the patients with p80 expression were younger than those without p80 expression (P < 0.05). The prognosis of the former was worse than in the later (P < 0.05). CONCLUSIONS: p80 positive ALCL could be a new clinicopathological entity. The polyclonal antibody anti-p80 might be an useful marker for the differential diagnosis and prognosis estimation of ALCL. PMID- 11866901 TI - [Inhibition of monocytes adhesion to the intima of arterial wall by local expression of antisense monocyte chemotactic protein-1]. AB - OBJECTIVE: To study the mechanism of monocyte recruitment in atherogenesis and to clarify the effect of monocyte chemotactic protein-1 (MCP-1) in this process. METHODS: Femoral arteries isolated from the rabbits which had been fed with a high cholesterol diet and locally perfused with MM-LDL within the artery beforehand, were used as the models. Antisense MCP-1cDNA was transferred into the arterial wall by injecting recombinant LNCX-anti-MCP-1/liposomal complex in the femoral sheath and the periarterial tissue. RESULTS: Expression of antisense MCP 1 mediated by recombinant LNCX plasmid/lipsomal complex gene transfer enabled to inhibit MCP-1 gene expression and adhesion of monocyte to the intima. CONCLUSION: MCP-1 plays an important role on the recruitment of monocytes in the arterial wall, which provides a potential clue in developing a gene therapy project for the prevention and treatment of atherogenesis. PMID- 11866900 TI - [Subcutaneous panniculitis-like T-cell lymphoma: expression of cytotoxic-granule associated protein TIA-1 and its relation with Epstein-Barr virus infection]. AB - OBJECTIVE: To study the clinicopathologic features, the expression of cytotoxic granule-associated protein TIA-1 and Epstein-Barr virus (EBV) infection in subcutaneous panniculitis-like T-cell lymphoma (SPTCL). METHODS: Totally 17 cases of SPTCL were studied. Immunohistochemical staining was done for TIA-1, CD45RO, CD3, CD20 and CD68, and in situ hybridization for EBV encoded small nuclear RNA (EBER1/2). RESULTS: In the 17 cases of SPTCL, the ratio of male-to-female was 1:1.1 and the median age was 24 years old. The main clinical manifestations were asymptomatic plaques, nodules and masses. Histologically, the pattern of lymphoid cells infiltration was panniculitis. The infiltrated lymphoid cells were pleomorphic and their mitotic figures were few (0 - 2/hpt). In the tumor tissue, there might be epithilioid granulomas, multinuclear cells, beanbag cells, small pieces of necrosis and small vessels with lymphoid cells infiltration in the vascular wall. Immunohistochemically, all the cases showed a T-cell phenotype and the expression rate of TIA-1 was 100% (16/16). By in situ hybridization, the positive rate of EBV-EBER1/2 was 29.4% (5/17). CONCLUSIONS: SPTCL has a specific histopathologic picture. It may originate from cytotoxic T lymphocytes or NK cells. Some of them may be accompanying with EBV latent infection. PMID- 11866902 TI - [The expression and effects of isoforms of macrophage colony stimulating factor in human leukemic cell lines]. AB - OBJECTIVE: To explore the expression and effects of isoforms of macrophage colony stimulating factor (M-CSF) in human leukemic cell lines. METHODS: Three normal human peripheral blood mononuclear cells (PBMCs) and 4 human myelomonocytic leukemic cell lines including J6-1, J6-2, K562 and HL-60 were studied using ABC immunoperoxidase assay, indirect immunofluorescence staining, flow cytometry, Western blot and reverse enzyme-linked DNA-protein interaction assay (reverse ELDIA). RESULTS: M-CSF was noticed to be localized in the cytoplasm, nucleus and at the cell membrane in 4 human leukemic cell lines; expression of M-CSF was not detected in normal human PBMCs without PHA stimulation. Human PBMCs stimulated by PHA expressed a low level of M-CSF. Frequencies of membrane bound M-CSF expression in J6-1, J6-2, K562 and HL-60 were 71.6%, 69.7%, 42.7% and 57.4% respectively. Frequencies of cytoplasm and nucleus associated M-CSF were 65.7%, 45.4%, 36.5% and 72.5% respectively. The cytosolic bound M-CSF was expressed in J6-1 cell as four isoforms with a molecular weight of 14,000, 16,000, 20,000 and 44,000. While nucleus associated M-CSF expressed as two isoforms with a molecular weight of 16,000 and 20,000. Anti-M-CSF monoclonal antibody could dramatically inhibit proliferation of leukemic cells and its inhibitory effect was related to the levels of membrane bound M-CSF expression in leukemic cells. Reverse ELDIA showed that M-CSF could bind with DNA in vitro. CONCLUSIONS: Expression of M-CSF isoforms is heterogeneous and polymorphous in leukemic cells. Membrane bound M CSF is crucial for the proliferation of leukemic cells, which might be a DNA bound protein and could be involved in the transformation and tumorigenesis of hematopoietic cells. PMID- 11866903 TI - [An experimental study on cerulenin induced apoptosis of human colonic cancer cells]. AB - OBJECTIVE: To investigate whether cerulenin could induce apoptosis and suppress the proliferation of human colonic cancer cells. METHODS: Cell morphology, microculture tetrazolium assay (MTT), agarose gel electrophoresis and flow cytometry assays were performed to identify the inhibitory effect on cell proliferation and the induction of apoptosis in human colonic cancer cells (LoVo). RESULTS: Cerulenin was able to induce LoVo cells death via apoptosis. Cerulenin also inhibited the proliferation of LoVo cells in vitro and was dose dependent. Nuclear DNA of apoptotic cells displayed ladder bands characteristic of internucleosomal DNA fragmentation and apoptotic peak. The inhibitory effect of cerulenin on proliferation of LoVo cells was associated with cell cycle arrest from S to G(2)-M phase transition and with induction of apoptosis. In contrast, cerulenin could not affect the proliferation of human fibroblasts in vitro. CONCLUSION: The fatty acid synthase inhibitor, cerulenin, enables to induce cell apoptosis and to suppress the growth of human colonic cancer cells by inhibition of the synthesized fatty acids endogenously in the cancer cells. PMID- 11866904 TI - [The expression of sialyl Lewis-X antigen in LoVo, HT29 cell lines of colorectal cancer and its correlation with metastatic potential]. AB - OBJECTIVE: To study the correlation of sialyl Lewis-X (SLeX) antigen expression with the metastatic potential of human colorectal cancer. METHODS: The expression of biosynthetic enzyme of SLeX antigen--alpha1, 3Fuc-T mRNA in various cell lines of colorectal cancer possessed with different metastatic potentials (LoVo and HT(29)) was investigated by in situ hybridization. The expression of SLeX antigen was studied using immunohistochemistry and fluorescence-activated flow cytometry qualitatively and quantitatively. RESULTS: The expression level of alpha1, 3Fuc-T mRNA and SLeX antigen in highly metastatic LoVo cells was high (21.2 +/- 7.7, 32.8 +/- 10.9, P < 0.05), and the expression level was low in HT(29) cells (10.8 +/- 5.2, 21.9 +/- 8.8) which are known possessed with a lower metastatic potential. CONCLUSION: The expression of SLeX antigen is correlated to the metastatic potentiality of human colorectal cancer. PMID- 11866905 TI - [The relationship between laminin-receptor and nm23 protein expression and its correlation with interstitial microvascular density and tumor metastasis in breast carcinoma]. AB - OBJECTIVE: To study the relationship between 67,000 laminin, laminin-receptor (LN R) and nm23 protein expression, interstitial microvascular density (MVD) and tumor metastasis in breast carcinomas. METHODS: The expression of laminin (LN), LN-R, FVIIIRAg and nm23 protein were detected in 73 cases of breast carcinoma with immunohistochemical technique and analyzed. RESULTS: A significant difference in LN expression was found in breast carcinoma according to their pathological grade. A positive relation exists between LN-R expression and lymph node metastasis. A positive relationship exists between the degree of LN-R and MVD expression. A significant difference in lymph node metastasis cases exists between nm23 protein positive expression group and negative expression group. When the expression of nm23 protein was inhibited, the degree of LN-R, MVD expression increased and positively related with lymph node metastasis. CONCLUSION: LN-R expression enhancement, microvascular density increase and nm23 expression inhibition may be the reliable markers for predicting tumor metastasis and prognosis. PMID- 11866906 TI - [The relationship between vascular endothelial growth factor, microvascular density, lymph node metastasis and prognosis of breast carcinoma]. AB - OBJECTIVE: To investigate the relationship between vascular endothelial growth factor (VEGF) microvascular density (MVD), lymph node metastasis and prognosis of breast carcinoma (BC). METHODS: VEGF protein expression and MVD in 92 cases of BC and VEGF mRNA expression in part of the cases were studied by immunohistochemistry methods and reverse-transcription polymerase chain reaction (RT-PCR) technique. RESULTS: VEGF mRNA expression in BC tumor tissues were higher than those in adjacent normal tissues. Increment of both VEGF(121) and VEGF(165) showed significant difference (P < 0.05 approximately 0.01). In addition, VEGF(145) expression was also observed in BC. There was a close positive correlation between VEGF and MVD (r = 0.702, P < 0.01). VEGF protein expression and MVD correlated significantly with lymph node metastasis and tumor relapse (P < 0.05 approximately 0.01). The time period of relapse-free-survival (RFS) in the patient group with high VEGF expression and MVD was significantly lower than RFS in the group with low VEGF expression and MVD (P < 0.01). CONCLUSION: VEGF is highly related to angiogenesis of BC. The increase of VEGF and MVD may promote lymph node metastasis and relapse of BC. VEGF and MVD may have prognostic value in RFS of BC patients. PMID- 11866907 TI - [Ultrastructural and immunohistochemical characteristics of pericytes during neovascularization in breast carcinoma]. AB - OBJECTIVE: To study the morphology of pericyte, the relationship between pericytes and neovascularization of breast carcinoma. METHODS: Ultrastructural, immunohistochemical and quantitative morphology analysis techniques were used to study 89 cases of human breast carcinoma. 4 human granulation tissues cases were used as control. RESULTS: The positive cells for FVIIIRAg were endothelial cells situated inside the vascular wall with abundant cytoplasm containing caveolae and Weibel-palade bodies. The positive cells for alpha-SMA were pericytes with cytoplasm containing abundant myofilament, and connections with endothelial cells. The pericyte-endothelial junctions can be found. Quantitative morphological analysis showed that the number of pericytes in the low microvascular density areas are much higher than that in the high- microvascular density areas (P < 0.0005). CONCLUSION: Ultrastructural observation and immunohistochemical staining of alpha-SMA are useful in identifying pericytes. The results of this study imply that pericytes may possibly inhibit angiogenesis. PMID- 11866908 TI - [Sarcomatoid carcinoma in breast]. AB - OBJECTIVE: To observe the distinctive pathomorphological features of sarcomatoid carcinoma (SC) in mammary gland as well as its diagnosis and differential diagnosis. METHODS: 1 538 malignant tumors of the breast were reviewed. 15 cases (0.98%) were diagnosed as SC. AE1/AE3, carcinoembryonic antigen (EMA), vimentin, S-100 protein, actin, estrogen receptor (ER) and progesterone (PR) were performed with SP immunohistochemical staining methods. RESULTS: The SC were divided into four morphological types according to the distinctive features of their sarcomatoid components. (1) Polymorphic sarcoma type: sarcomatoid components are polymorphic; (2) Spindle cell type: sarcomatoid components consisted of spindle cells; (3) Osteoclastic giant cell type: a huge number of osteoclastic giant cells in the sarcomatoid components; (4) Myxocartilage type: myxocartilaginoid components are seen in the tumor tissue. With immunohistochemical staining, the expression rates of AE1/AE3, EMA, vimentin, S-100 protein and actins in the epithelial components were in 11, 15, 11, 12 and 8 cases respectively, and sarcomatoid components were in 10, 13, 13, 13 and 10 cases respectively, while ER and PR were positive only in 2 cases. CONCLUSION: Sarcomatoid carcinoma of the breast should be distinguished from carcinosarcoma, desmoplastic carcinoma, sarcoma, nodular fasciitis and fibromatosis. PMID- 11866910 TI - [The inhibitory effect of antisense adenovirus vector of telomerase on telomerase activity in breast cancer cells MCF-7]. AB - OBJECTIVE: To inhibit telomerase activity in breast cancer cells MCF-7 by antisense ribonucleotide technique. METHODS: Recombinant adenovirus was chosen as gene delivery vector to transfer and express the antisense RNA component of telomerase in tumor cells. The recombinant adenovirus of anti-telomerase RNA was obtained through gene recombination and co-transfection with lipofectin. The integrated potential of antisense virus was certified by Southern blot and the telomerase activity of MCF-7 was analysed by TRAP-silver staining. RESULTS: The MCF-7 cell line is a typical model for malignant breast cancer. In this study, the telomerase activity of MCF-7 was significantly inhibited after antisense virus infection when compared with the control cell lines. CONCLUSION: The introduction of antisense cDNA of telomerase RNA can significantly inhibit the activity of telomerase in MCF-7 cells. PMID- 11866909 TI - [Endometrial nuclear progesterone receptors in infertile women]. AB - OBJECTIVE: To investigate the endometrial nuclear progesterone receptors (PgR) as well as its relationship with retarded endometrial development (RED) in infertile women. METHODS: The progesterone profile, endometrial nuclear progesterone receptors (PgR) and endometrial biopsies were studied by using (125)I radioimmunoassay and immunohistochemical technique in 53 infertile women during median luteal phase. When the endometrial dating lagged 2 days behind the dating as determined by luteinizing hormone (LH) surge, diagnosis of luteal phase defect (LPD) was made. According to progesterone profile and endometrial morphology, the patients were divided into three groups: normal, luteal phase defect (LPD) and pseudocorpus luteum insufficiency (PLI). RESULTS: During median luteal phase, RED were found in all of 24 cases with LPD and PLI, but PgR contents were different in them. Mean progesterone profile of 9 cases of PLI cycles was normal (60.2 nmol/L) but the endometrial nuclear PgR appeared to be prominently lower than in normal cycles during the median luteal phase (P < 0.01). Average progesterone level of 15 cases of LPD cycles were significantly lower (29.9 nmol/L) but endometrial nuclear PgR were similar to normal group (P > 0.05). CONCLUSION: Retarded endometrial development may have different causes. Measurement of endometrial nuclear PgR is of assistance in differential diagnosis. PMID- 11866911 TI - [The effects of cyclin E on the growth and other cell cycle related genes of breast carcinoma cells MCF-7]. AB - OBJECTIVE: To investigate the effects of cyclin E in high and low expression on the cell growth and other cell cycle associated genes of MCF-7 cells. METHODS: Eukaryotic expression vehicles of sense and antisense cyclin E were constructed and transferred into the MCF-7 cell line using lipofectAMINE. The integration and expression of cyclin E were conformed by Southern and Western blot. The cells growth was observed and the changes in cell cycle were analyzed by flow cytometry. The expression of other cell cycle associated genes was assayed using Western blot. RESULTS: A high expression of cyclin E enabled to promote the cell growth and DNA synthesis and accelerated the proceeding of G(1) phase to S phase, It also promoted the phosphorylatin of pRB and up-regulate the expression of p27 while a low expression of cyclin E showed an opposite effect. CONCLUSION: Different expression of cyclin E enables to affect growth of MCF-7 cell by the changes of cell cycle related genes. PMID- 11866912 TI - [Effects of high-affinity nerve growth factor (NGF) receptor gene on NGF-induced differentiation of neuroblastoma cell line]. AB - OBJECTIVE: To study the effects of transfection of high-affinity nerve growth factor receptor gene (trkA) on NGF-induced differentiation of human neuroblastoma cell line IMR-32. METHODS: The recombinant retrovirus vector containing exogeneous trkA gene was constructed and packed by PA317 packaging cell line. The neuroblastoma cell line IMR-32 was transfected by virus containing supernatant. The transformant cell line was confirmed by Southern blot and RT-PCR techniques. The NGF was used to induce cellular differentiation of the transformant cells. RESULTS: The trkA gene was successfully transferred and expressed in the neuroblastoma cells. After NGF treatment, the transformant cells displayed apparent neuron-like differentiation morphologically, and a slower rate of cell growth (MTT value 0.028 +/- 0.003) compared with original cell line (0.258 +/- 0.017) and empty virus transformed cell line (0.237 +/- 0.011). The cells remained in differentiated status after withdrawing the NGF from the medium. The transformant tumor cells rarely formed colonies in soft agar and failed to form tumor in nude mice. CONCLUSION: Restoration of high-affinity NGF receptor (trkA) expression in neuroblastoma cells could induce non-reversal differentiation. The trkA might be the important factor during NGF-induced differentiation of neuroblastoma cell. PMID- 11866913 TI - [Morphological changes of mitochondria in apoptosis of esophageal carcinoma cells induced by As(2)O(3)]. AB - OBJECTIVE: In order to make clear how As(2)O(3) targets the cells of esophageal carcinoma, the morphology of mitochondria and expression of bcl-2/bax were selected to study the SHEEC1 cell line during early apoptosis. METHODS: The esophageal carcinoma cell line SHEEC1 was treated by As(2)O(3) with concentrations of 1, 2 and 3 micromol/L for the periods of 2, 4, 6, 12 and 24 h. Annexin-V-Fluorescein and DNA histogram were used to detect the early apoptotic cells by flow cytometry. The morphologic changes of the apoptotic cells were examined by means of light and electron microscopy. Changes in mitochondria of living cells were detected with Rhodamin 123 fluorescent probe by fluorescent microscopy. The expression of bcl-2 and bax in floating cells were determined by immunohistochemical procedures. RESULTS: Typical apoptotic morphological changes were found in SHEEC1 at 24 h after As(2)O(3) treatment. Cells in the early stage of apoptosis were detected by Annexin-V labeled at 4 h. The ultrastructural alterations of mitochondria were found as following: within 4 h after administrating As(2)O(3), mitochondrial hyperplasia with small electron-dense materials in its matrix was the earliest manifestation of cell injury. At 6 h progressive mitochondrial swelling lead to balloon-like appearance and its outer membrane disrupted at 12 h. In all groups, down regulated expression of bcl-2 and over expression of bax were always found in damaged cells. CONCLUSION: In the SHEEC1 cell line morphological changes of mitochondria are the early events in apoptosis induced by As(2)O(3) and apoptosis is closely related to over expression of bax and down-regulating expression of bcl-2. It is possible that As(2)O(3) is a mitochondriotoxic anticancer agent for esophageal carcinoma. PMID- 11866914 TI - [Inhibition of hTR activity for suppression of lung cancer cell proliferation using antisense phosphorothioate oligonucleotides]. AB - OBJECTIVE: Telomerase activity was measured in 8 lung tumor cell lines and transfected antisense phosphorothiate oligodeoxynucleotides (anti-PS-ODN) of hTR as therapeutic agents into target cell LTEP-a-2 to investigate the inhibitory effect on telomerase activity and tumor cell growth. METHODS: 1. Telomerase activity assay in 8 human lung tumor cell lines using telomerase PCR ELISA. 2. Synthesized anti-PS-ODNs of hTR and random PS-ODN transfected with Clonfectin into LTEP-a-2 cell lines for 72 hr. 3. Measure telomerase activity by telomerase PCR ELISA, SDH activity by MTT assay and cell growth. RESULTS: Eight cell lines showed positive expression of telomerase activity. Various anti-PS-ODNs could inhibit telomerase activity, SDH activity and cell growth. The inhibition became more marked with the increase of anti-PS-ODNs concentrations. Concentrations of 5 - 40 micromol/L anti-PS-ODNs of hTR specifically reduced the telomerase activity by 25.7% - 84.0%. PS-ODNs of anti-hTR were also able to reduce SDH activity by 19.4% - 74.7% at 5 - 40 micromol/L. The dose of 5 - 40 micromol/L PS-ODNs of anti hTR had the ability to inhibit cell growth by 14.8% - 72.5%. The results indicate that random sequence of PS-ODN(9) failed to inhibit telomerase, SDH activity and cell growth. A statistically significant difference exists between random PS-ODN and three anti-PS-ODNs (P < 0.01). CONCLUSIONS: Human lung tumor cell lines express high telomerase activity. PS-ODNs of anti-hTR had the ability to inhibit telomerase and reduce LTEP-a-2 cell growth and metabolism. PMID- 11866915 TI - [The role of bFGF in renal tubular injury and the interstitial lesion]. AB - OBJECTIVE: The role of bFGF in renal tubular injury, regeneration and interstitial lesion of rats was studied. METHODS: The renal tubular injury model of rat was induced by intraperitoneal injection of gentamicin. The expression of bFGF and FGFR1 was examined by Northern blot, in situ hybridization and immunohistochemistry. (3)H-TdR incorporation was used to detect the role of bFGF in promoting the proliferation of renal tubular epithelial cells and renal fibroblasts. RESULTS: The expression of bFGF mRNA and protein synthesis of bFGF and FGFR1 increased during the process of injury and regeneration of tubular epithelial cells. In contrast, normal tubular epithelium failed to express bFGF, except renal interstitial fibroblasts and monocytes/macrophages. bFGF promoted proliferation of renal tubular epithelium as well as renal interstitial fibroblasts. CONCLUSION: bFGF is secreted by tubular epithelial cells damaged by nephrotoxicity of gentamicin, and FGFR1 expressed in damaged renal tubules and interstitial fibroblasts. bFGF not only can promote tubular epithelia to proliferate, but can also stimulate interstitial fibroblasts to proliferate. Correlation exists between bFGF, renal tubular injury and interstitial lesions. PMID- 11866916 TI - [A study on the mechanism of anisodamine against thrombosis]. AB - OBJECTIVE: To investigate the effects and the mechanism of Chinese herb anisodamine on PAI-1 expression in LPS stimulated endothelial cells (ECs). METHODS: Human umbilical vein endothelial cells (HUVECs) were cultured by trypsin digestion method. PAI-1 protein in HUVEC conditioned medium was measured by sandwich enzyme-linked immunosorbent assay (ELISA), and PAI-1 mRNA expression in HUVEC was determined by Northern blot analysis. HUVEC nuclear factor-kappa B (NF kappaB) nuclear translocation was assessed by immunocytochemistry technique. RESULTS: LPS treatment of cultured HUVECs resulted in a significant increase of PAI-1 protein as well as mRNA expression by these cells. However, when HUVECs were incubated with LPS plus anisodamine, the upregulation of PAI-1 by LPS was significantly abated. The expression of PAI-1 protein and mRNA was decreased by 30% and 24% respectively compared with LPS treatment alone. Moreover, anisodamine was able to decrease the basal level of PAI-1 protein and mRNA by 74% and 70% respectively. Furthermore, anisodamine could block LPS induced NF-kappaB of HUVECs nuclear translocation. CONCLUSION: Anisodamine inhibits both basal and LPS induced PAI-1 protein as well as mRNA expression in ECs. The inhibition of anisodamine on LPS-induced PAI-1 in ECs may take place via the NF-kappaB pathway. PMID- 11866917 TI - [Clinicopathologic study of angiogenesis in human hepatocellular carcinoma]. AB - OBJECTIVE: To study clinicopathologic significance of microvessel density (MVD) and the expression of vascular endothelial growth factor (VEGF) in human hepato cellular carcinoma (HCC). METHODS: 30 HCC cases with neighboring noncancerous tissues, 10 liver cirrhosis cases, 5 mild chronic hepatitis cases and 4 normal liver tissue samples were stained with CD34, VEGF by immunohistochemistry method. Quantitation of sinusoid-like vessels and semi-quantitation of VEGF expression were performed and analyzed in conjunction with the clinico-pathological features of HCC. RESULTS: All HCC tissues showed diffuse CD34 expression and its site of distribution was somewhat corresponding to the original structure of sinusoid like pattern, while CD34 was negative in the "sinusoid" regions of normal and chronic liver disease. Both MVD and VEGF were correlated to portal vein invasion of HCC, MVD was also correlated to the tumor size. The MVD of tumor less than 2 cm in diameter was significantly lower than that of tumors with a diameter of 2 similar 5 cm. The positive rate of VEGF in HCC was 83.3%. The expression of VEGF was also remarkable in the neighboring noncancerous tissues in which inflammatory cell infiltration was apparent. CONCLUSIONS: MVD and expression of VEGF were remarkably increased in HCC. Both were correlated with portal vein invasion. VEGF derived from tumor cells and noncancerous tissues may be associated with the angiogenesis of HCC. PMID- 11866918 TI - [Morphologic analysis of 14 cases of hepatic angiomyolipoma]. AB - OBJECTIVE: To study the morphology of hepatic angiomyolipoma (HAML). METHODS: HE staining, PAS staining, immunohistochemistry and electronic microscopy were used to study 10 cases from our department and 4 cases from consultation. RESULTS: All 14 HAML cases showed perivascular epithelioid cells (PEC) with 4 different types of morphology. They presented characteristic PAS positivity premelanosome like structures and dense body under electronic microscopy. Immunohistochemistry demonstrated positive reaction for both actin and HMB45. CONCLUSION: Light and electronmicroscopic features and immunophenotype of HAML are characteristic of PEC related lesion. PMID- 11866919 TI - [Detection of alterations of the DPC4 gene in paraffin-embedded tissues of pancreatic carcinomas]. AB - OBJECTIVE: To examine the alteration and significance of the DPC4 gene in paraffin-embedded tissues of pancreatic carcinomas. METHODS: Polymerase chain reaction and single-strand conformation polymorphism analysis were used to search for deletions and mutations in the DPC4 gene in 46 cases of pancreatic carcinomas. RESULTS: Thirteen of forty-six (28.3%) cases were found to have homozygous deletions in exon 1, 2, 3, 4, 8 and 11. One was in exon 11, one in exon 1 and 11, one in exon 2 and 3, one in exon 3 and 8, one in exon 1, 2 and 8, one in exon 2, 4 and 11, one in exon 3, 4 and 11, three in exon 3, 4 and 8, one in exon 2, 3, 4, and 8, one in exon 2, 3, 8 and 11, one in exon 2, 3, 4, 8 and 11. Intragenic mutations were found in 10 of 46 cases (21.7%). One case was in exon 1, one in exon 2, three in exon 8, four in exon 11, and one in exon 4 and 11. The total frequency of intragenic changes of DPC4 in paraffin-embedded tissues was 45.6% (21/46). CONCLUSION: Inactivation of tumor-suppressor gene DPC4 may play an important role during the tumorigenesis of pancreatic carcinomas. PMID- 11866920 TI - [Significance of fucose expression in lung carcinoma and their brain metastases]. AB - OBJECTIVE: To investigate the expression of fucose in brain metastases and their primary tumors. METHODS: Affinity histochemistry method was used to detect the expression of fucose in lung carcinoma and their brain metastases of 39 cases with available survival data. RESULTS: Of the 39 cases, 21 cases were positive for expression of fucose and 12 cases were negative both in the primary tumor and their metastases. 2 cases showed positive only in primary tumors and 4 cases only in metastatic tumors. The positive intensity (Dm) and percentage of the positive area (Sm) of fucose in brain metastases were stronger and higher than in their primary cancers (Dp, Sp), both were statistically significant (P = 0.0015 and P = 0.001 respectively). Multivariate analysis showed that the percentage of the positive area of the primary tumor and the positive intensity of the brain metastases was the significant predictor of survival after manifestation of the brain metastasis (Tm). CONCLUSION: Adenocarcinomas of the lung were prone to metastasize to the brain compared with other types of lung cancer. The Dm and Sm of fucose in brain metastasis was significantly stronger and higher than that of their primary tumor. The outcome of patients with brain metastasis could be predicated by Sp and Dm. Surgical resection of the primary lung cancer and their brain metastasis(es) resulted in better prognosis. PMID- 11866921 TI - [Matrix metalloproteinase-1 and coronary atheroslerotic plaque rupture]. AB - OBJECTIVE: To investigate the relationship between matrix metalloproteinase-1 (MMP-1) and coronary atherosclerotic plaque rupture, and the cellular source of MMP-1 within the plaques. METHODS: 42 cases, among which 20 died of acute myocardial infarction, 10 with unstable angina history and 12 with stable angina history but died of other diseases, were selected. All the branch of coronary arteries were examined, parts of the segments were selected for immunohistochemical staining, 5 markers against alpha-smooth muscle actin, CD20, CD45RO, CD68 and MMP-1 were performed. RESULTS: Plaque rupture and thrombosis were found in almost all the cases of acute myocardial infarction and unstable angina. But in the cases of stable angina, the majority of the plaques were stable ones. The expression of MMP-1 in the ruptured plaques were stronger than the unruptured ones (t = -8.07, P < 0.05); Positive relationship was also noted between the expression of CD68 and MMP-1 (r = 0.75, P < 0.05). CONCLUSION: Macrophages are capable of degrading extracellular matrix by secreting MMP-1; Enhanced secretion of MMP-1 within the coronary atherosclerotic plaques has significant relationship with plaque rupture. PMID- 11866922 TI - [A histopathologic and immunohistochemical study of prognostic factors in esophageal squamous cell carcinoma]. AB - OBJECTIVE: To investigate the prognostic value of various histopathological features and expression of immuno-phenotypes in esophageal squamous cell carcinoma (ESCC). METHODS: The tumor samples from 97 primary ESCC patients with potentially curative resection were evaluated for the following parameters: tumor size, grade of differentiation, pattern of invasion, depth of invasion, degree of lymphocyte infiltration, lymph node metastasis, TNM stage and p53, EGFR, c-erbB 2, nm23, cathepsin D protein expression. The prognostic impact of these parameters were analyzed by univariate and multivariate survival analyses. RESULTS: In the univariate Kaplan Meier analysis, poor differentiation, infiltrating growth pattern, invasion reaching adventitia, absent lymphocytic infiltration, lymph node metastasis, high staging, EGFR positive expression and nm23 negative expression were all associated with lower survival rate (Log-rank test, P < 0.05, respectively). However, the multivariate Cox analysis demonstrated that only pattern of invasion (RR = 2.000), degree of lymphocyte infiltration (RR = 0.509), TNM stage (RR = 1.733), EGFR expression (RR = 1.812) and c-erbB-2 expression (RR = 2.364) were significant prognostic factors (P < 0.05, respectively). CONCLUSION: For ESCC, the pattern of invasion, degree of lymphocyte infiltration, EGFR and c-erbB-2 protein expression had independent prognostic values as well as the TNM stage. Comprehensive analysis of these parameters can be helpful to identify biological behavior of ESCC and predict more accurately the prognosis of patients with ESCC. PMID- 11866923 TI - [Study on the needle biopsies of 100 prostatic adenocarcinoma]. AB - OBJECTIVE: To study the morphological diagnostic criteria of prostatic adenocarcinoma in needle biopsy. METHODS: 250 cases of prostatic needle biopsy including 100 cases of adenocarcinoma were observed retrospectively. The diagnostic significance of 20 parameters of morphological criteria including the architectural changes, cytologic features, invasion and some additional histologic features (intraepithelial neoplasm, intraluminal crystelloid body, intraluminal basophilic mucin secretion, collagen nodule and coagulative necrosis) were analyzed. RESULTS: All 20 parameters of morphological criteria had practical significance in pathologic diagnosis, the most significant ones being architectural changes, prominent nucleoli, perineural and stromal invasion. CONCLUSION: The diagnosis of prostatic adenocarcinoma is based on a constellation of cytologic and architectural features. PMID- 11866924 TI - [Zymographic analysis of matrix metalloproteinases in mouse carcinoma cell lines with different lymphatic metastasis potential]. AB - OBJECTIVE: To investigate the correlation between lymphatic metastasis and matrix metalloproteinases (MMPs) activity. METHODS: Zymographic analysis was used to detect the difference of MMPs product and activities between high lymphatic metastatic cell line (HCa-F) and low lymphatic metastatic cell line (HCa-P), that were cultured in the medium including homogenate of lymph node or liver or spleen tissue. RESULTS: HCa-F and HCa-P cells produced small quantities of MMP-9 in 1640 medium. After adding lymph node homogenate to the culture medium, F cells produced a large quantity of MMP-9 as well as active MMP-9 and MMP-2. P cells also produced MMP-9, active MMP-9 and MMP-2, but were much lower than that of F cells. Neither F cells nor P cells produced MMPs in the media with homogenate of liver or spleen added. CONCLUSION: The metastatic potential of F cells is stronger than that of P cells may be due to F cell's ability to secrete MMPs is stronger than P cell's, therefore the production of MMPs of hepatocarcinoma cells is closely correlated to their metastatic potential. PMID- 11866925 TI - [Study on the modulation of the inflammatory response in mouse hepatic vasculitis with sodium selenite and vitamin E antioxidants]. AB - OBJECTIVE: To identify the regulatory effect of sodium selenite and vitamin E on complement-neutrophil-reactive oxygen species (ROS)-activation feedback mechanism mediated inflammatory response. METHODS: The modulation of inflammatory response through the complement-neutrophil-ROS-activation feedback cycle with sodium selenite and alpha-tocopherol was verified both in vitro tests by chemiluminescense technique and complement fixation to detect ROS production and complement activation as well as in vivo mouse hepatic vasculitis models to test for the regulation of the inflammatory response. RESULTS: Convincing results were observed as the incidence of hepatic vasculitis dropped from 100% in the control group down to 20% in the seleno-antioxidant vitamin E treated group, indicating down regulation of the inflammatory response. CONCLUSIONS: Elucidation of the mechanism of complement mediated inflammatory response is a promising area for new therapeutic developments in the modulation of inflammatory response. This study suggests that selenium, vitamin E and other antioxidants may be the useful therapeutic agents in those disorders in which upregulation of inflammatory response has been implicated, e.g. ischemia, reperfusion injury, severe sepsis, rheumatoid arthritis and hepatitis. PMID- 11866926 TI - [The effect of TGF-beta1 on the expression of plasminogen activator inhibitors 1 and matrix metalloproteinases in cultured rat mesangial cells transfected with antisense-TGF-beta1 vector]. AB - OBJECTIVE: To study the plasminogen activator inhibitors 1 (PAI-1) and matrix metalloproteinases (MMPs) expression on the cultured rat mesangial cells (MsC) transfected with antisense-TGF-beta1 vector. METHODS: Lipofectin method was used to transfect antisense-TGF-beta1 vector into MsC, Western blot and Northern blot analysis for detecting TGF-beta1 peptides and antisense TGF-beta1 RNA level. The expression of PAI-1 and MMPs were determined by Northern blot, Western bolt and zymography assay. RESULTS: The MsC clone (MAT1) which blocked the expression of TGF-beta1 peptides was successfully established. The MsC clone showed increased expression of MMP-2 mRNA protein and enzyme activity of MMP-2, 3, 7. However, expression of PAI-1 mRNA of MsC clone was suppressed. CONCLUSION: Blocked expression of TGF-beta1 can upregulate the expression of MMP-2, 3, 7 and downregulate the expression of PAI-1 mRNA in MsC. It is possible that antisense TGF-beta1 may be used for gene therapy in glomerular diseases. PMID- 11866927 TI - [The expression of connexin 43 and desmin in viral myocarditis]. AB - OBJECTIVE: To study the cytobiological basis of constriction dysfunction and arrhythmia in cardiac muscle cells with viral myocarditis. METHODS: The expression of connexin 43 and desmin in cardiac muscle cells of mice with experimental viral myocarditis was determined by immunohistochemistry. RESULTS: In normal mice, connexin 43 and desmin are located in the intercalated disks of cardiac muscle, and the latter also revealed a positive immunoreactivity in the cross striations of sarcomeres. In viral myocarditis, the expression of both became much weakened, can even become negative. CONCLUSION: In myocarditis, expression of connexin 43 and desmin in the involved cardiac muscle cells was inhibited, resulting in dysfunction of gap junctional communication and arrhythmia. PMID- 11866928 TI - [Expression and clinical significance of p53 protein and p21(WAF1) in non-small cell lung carcinoma]. AB - OBJECTIVE: To investigate the relationships between expression of tumor suppressor gene p53 protein, p21(WAF1), clinicopathology and prognosis in human non-small-cell lung carcinoma (NSCLC). METHODS: SP immunohistochemistry performed in all cases. RESULTS: One hundred forty-seven resected NSCLC were examined. There were 63 squamous cell carcinomas, 66 adenocarcinomas, 14 adenosquamous carcinomas and 4 large cell carcinomas. Overall 90 (61.2%) of the 147 cases had positive p53 staining. The positive ratios of p53 overexpression of squamous cell carcinomas, adenocarcinomas, adenosquamous carcinomas and large cell carcinomas were 63.5% (40/63), 57.6% (38/66), 71.4% (10/14) and 50% (2/4) respectively. The average positive ratio of p21(WAF1) expression was 40.1% (59/147) in lung carcinoma. The positive ratios of p21(WAF1) expression of squamous cell carcinomas, adenocarcinomas, adenosquamous carcinomas and large cell carcinomas were 41.3% (26/63), 42.4% (28/66), 28.6% (4/14) and 25% (1/4) respectively. Over expression of p53 protein was associated with prognosis of the patients with adenocarcinomas. The positive ratio of p53 expression in over 3 year survival patients was 75% (21/28) and in less than 3 year survival, 44.7% (17/38) (P < 0.025). The positive expression of p21(WAF1) was related to prognosis of NSCLC. The ratio of 3-year survival (64.4%) in p21(WAF1) positive NSCLC was much higher than in p21(WAF1) negative patients (46.6%) (P < 0.05). Prognosis of patients with p53 positive but p21(WAF1) negative expression was poorer than those with p53 negative and p21(WAF1) positive NSCLC (P < 0.01). CONCLUSION: These findings suggest that over expression of p53 in adenocarcinomas could be used as a prognostic factor for patients. p21(WAF1) is of protein has a definite value in judging the prognosis in NSCLC. Combined expression of p53 and p21(WAF1) can be used as the prognostic markers in NSCLC and are of significance in estimating the prognosis of patients. PMID- 11866929 TI - [The morphological characteristics of desmoplastic fibroblastoma]. AB - OBJECTIVE: To study the clinical and pathological characteristics of desmoplastic fibroblastoma. METHODS: Three cases of desmoplastic fibroblastoma were evaluated by light microscopy and immunohistochemistry. RESULTS: The patients were all adults, 2 males and 1 female, age range 46 - 62 years (mean 53 years). Clinically, the tumors presented as slowly growing painless masses, which were located in the deep soft tissue of the lower extremity, the subcutaneous tissue of the forearm and the ankle respectively. Grossly, they were well-circumscribed neoplasms with a firm consistency. Histologically, the tumors were composed of paucicellular spindle or stellate-shaped fibroblastic cells embedded in a densely fibrous or fibromyxoid stroma. The neoplastic fibroblastic cells resembled the reactive fibroblasts by containing fine chromatin and small distinct nucleoli without cellular atypia or mitotic figures. Immunohistochemically, the neoplastic cells showed diffuse and strong immunoreactivity for vimentin, focal staining for alpha-SMA, MSA and desmin. CONCLUSION: Desmoplastic fibroblastoma is a benign fibroblastic neoplasm with distinctive clinical and pathological characteristics. It should be differentiated from a variety of reactive or low-grade fibroblastic lesions. PMID- 11866930 TI - [A clinicopathological study of hepatic focal nodular hyperplasia]. AB - OBJECTIVE: To study the clinicopathological features of focal nodular hyperplasia (FNH). METHODS: The clinicopathological characteristics of 25 cases of FNH were studied retrospectively. 20 cases followed for 6 to 47 months. All were evaluated by use of paraffin-embedded sections, special and immunohistochemical staining (EnVision method) and electron microscope. RESULTS: 17 female and 8 male FNH patients aged 14 to 58 (median 38) years of age, all alpha-fetoprotein negative, asymptomatic and normal biochemical liver tests in most cases. The macroscopic hallmark is a central stellate fibrotic scar, composed of fibrous connective tissue and tortuous blood vessels, the fibrous tissue radiated peripherally, dividing the mass into multiple, variably sized nodules, simulating the pattern of cirrhosis. Microscopically, multinodular proliferation of benign-appearing hepatocytes separated by bile-duct-containing fibrous septae that radiate from the central scar. Internodular bile duct proliferation is abundant and merge imperceptibly with the hepatocyte elements near the fibrous septa. Immunohistochemically, the keratin expessed by hepatocytes and by the proliferated bile ducts were similar. Electron microscopic examination found that the ultrastructure of tumor cells were similar to normal hepatocytes. 20 cases were followed for 6-47 months, all survived with no recurrence. CONCLUSION: FNH is a benign hepatocytic lesion, i.e. a reactive proliferation of hepatic cells to local blood vessel anomalies. PMID- 11866931 TI - [Histological diagnosis of regional lymph node micrometastasis with or without stromal reaction in gastric carcinoma]. AB - OBJECTIVE: To define lymph node micrometastasis and to investigate the difference between micrometastasis and microinvolvement. METHODS: A total of 1,087 regional lymph nodes were collected from surgical specimens of 17 patients with gastric carcinoma. 71 nodes were found to contain microlesions, 60 sections per node) and immunohistochemistical staining for CK18, EMA and SAM. Another 20 nodes containing macrometastasis were also studied. RESULTS: After serial sections, the microlesions disappeared in 34% (24 nodes), increased in 49% (35 nodes), microlesions transformed into macrometastasis in 17% (12 nodes). Microlesions presented as a cluster of tumor cells ( 75 g alcohol/day. A tendency of a mild persistent elevation of plasma HDL-C and apoA I was noticed corresponding to an increase of the amount of alcohol intake, and with no evidence of threshold effect observed in the samples of both the Yi-farmers and the Han people. Similar phenomenon was obtained in the sample of Yi-emigrants, but no statistical significance. The frequencies of the A allele of all 3 samples were similar. In Han and Yi-emigrant samples, the drinkers with genotypes of GG had a higher plasma HDL-C and apoA I level than that of the non-drinkers with the same genotypes. Drinkers with A allele had a lower plasma HDL-C and apoA I level than that of drinkers without A allele, and the non-drinkers with A allele had a higher levels of apoA I than of non-drinkers with genotypes of GG. It is estimated that 18% of the variability of plasma apoA I level could be explained by the G/A polymorphism in non-drinkers of Yi-emigrants (F = 8.94, P < 0.01). CONCLUSIONS: Current data suggest that a moderate alcohol consumption or a G to A substitution could make a lower incidence of CHD, but the beneficial effect of one will be negated by the other when both factors occur simultaneously. This finding is seemed valuable for a further study on the effect of the environmental factor or genetic factor in effecting the plasma apoA I level afterwards. PMID- 11866933 TI - [Aortic calcification and matrix metalloproteinases]. AB - OBJECTIVE: To study the relationship between arterial calcification and matrix metalloproteinases including MMP-1 and MMP-3. METHODS: 15 human aorta specimens from autopsies were stained with HE to identify calcification. Immunohistochemical method was used to determine the secretion of MMP-1 and MMP 3. RESULTS: Among the 15 aorta specimens, calcification was found in 10 and no calcification in the remaining 5. There was no significant difference in the percentage of MMP-1 positive cells between two groups, with 83.8% +/- 5.2% in calcified specimens vs 84.0% +/- 7.5% in non calcified ones. 9 of the 10 calcified aortas were positive for MMP-3 staining. Serial sections showed that the localization of positivity was in concordance with calcified regions. The 5 normal aortas were MMP-3 negative. The difference between the two groups was significant. CONCLUSION: MMP-3 secretion relates closely with arterial calcification. And MMP-3 may participate in calcium minerali-zation in arteries. MMP-1 has no relation with arterial calcification. PMID- 11866934 TI - [HPV16 E6E7 fragments transform immortalized human bronchial epithelial cells into neoplastic cells]. AB - OBJECTIVE: To study the effects of HPV16 E6E7 fragments on the biological behavior of immortalized human bronchial epithelial cells TR. METHODS: E6E7 fragments were constructed into retrovirus expression vector and then transfected into TR cells to observe changes in growth character and tumorigenesis in nude mice. IP-Western blots were done to analyze function of p27 protein as well as expression and phosphorylation of FAK and paxillin. RESULTS: Southern, Northern, Western blot confirmed the stable expression of E6E7 fragments in puromycin resistant cell strain TR/E6E7. TR/E6E7 proliferated faster, showed stronger growth ability in soft agar and eventually formed tumor after being inoculated into nude mice. IP-Western blot also showed sequestering of p27 protein from cyclin E/CDK2 complex, which implied increased activity of cyclin E/CDK2 and promoting G1 to S cell cycle. CONCLUSION: TR/E6E7 cells formed tumor in nude mice indicating that HPV infection played a role in pulmonary carcinogenesis. Sequestering of p27 protein from cyclin E/CDK2 complex may be responsible for increased proliferation in cells transfected by HPV16 E6E7. PMID- 11866935 TI - [Effects of adenovirus-mediated p16 and p53 genes transfer on apoptosis and cell cycle of lung carcinoma cells]. AB - OBJECTIVE: To explore the synergistic inhibition effect and apoptosis induction of p16 and p53 genes on lung carcinoma cells. METHODS: E1-deficient and replication-defective recombinant p16 and p53 adenoviruses were generated by liposome-mediated co-transfection of recombinant plasmid pAdCMV-p16 or pAdCMV-p53 along with pJM17 and homologous recombination in 293 packaging cell. The lung cancer cell line H358, which had a homozygous deletion of p53 gene and no expression of p16 mRNA and protein, was infected with recombinant p16 and p53 adenovirus either individually or together. RESULTS: Immunohistochemical analysis showed that recombinant adenovirus could transfer p53 gene into tumor cell with 98% efficiency. Western blot indicated that p16 and p53 proteins were expressed at a high level in infected H358 cell. Inhibition effect of p53 gene on proliferation of H358 cell was weaker than that of p16 gene, and the combined use of both genes could completely prevent the proliferation of H358 cell. In situ end-labeling and flow cytometry indicated that p16 could result in G(1) arrest of cell cycle and did not induce H358 cells to undergo apoptosis; p53 also induced apoptosis of few cells besides G(1) arrest; and the simultaneous use of p16 and p53 genes could induce marked apoptosis of H358 cells. CONCLUSION: p16 and p53 genes possess the synergistic inhibiting effect on growth of lung cancer cells and can cooperate to induce apoptosis of H358 cell. The combined application of recombinant p16 and p53 adenoviruses can be used as a new strategy for cancer gene therapy. PMID- 11866936 TI - [Effect of mouse p53 minigene on lung cancer cells with different 172 structures regulated by tetracycline]. AB - OBJECTIVE: To investigate the effect of mouse 172 wild-type p53 (Arg), pseudo wild-mutant-type p53 (Arg-->Leu) and mutant-type p53 (Arg-->His) induced by absence of tetracycline on the growth of PG cell line. METHODS: Three variant types of p53 minigene were sub-cloned by gene recombination into an expression vector which was controlled by tetracycline. Through LipofectAMINE, the vectors were transfected into p53 defective PG (248CGG-->CTT) cells, and the transfectants were screened in the selecting medium containing puromycin. Tumor suppressing effects were studied by MTT absorption, flow cytometry and Western blotting. RESULTS: Wild-type p53 and pseudo-wild-mutant-type p53 could lead cells to decrease their growth rates, arrest cell cycle and transactivation of p21(WAF1). Mutant-type p53 was defective in tumor suppression. CONCLUSION: Wild type p53 and pseudo-wild-type p53 may inhibit cell growth and induce cell cycle arrest. Some p53 variants such as 172 Arg-->Leu can still retain the tumor suppression function of the wild-type. PMID- 11866937 TI - [Inhibition of growth and metastasis of human giant cell carcinoma of the lung by transfection of antisense VEGF121 cDNA]. AB - OBJECTIVE: To explore the effects of blocking the VEGF/VEGF receptor paracrine pathway on growth and metastasis of human lung carcinoma cell line PG and to evaluate its potential application in gene therapy of cancer. METHODS: The eukaryotic expression vectors bearing either sense-VEGF121 cDNA or antisense VEGF121 cDNA was constructed and transfected into PG cells. In vitro and in vivo tests such as Northern blotting hybridization, Western blotting immunochemistry analysis, as well as xenografting in nude mice were used to analyze the effect of antisense-VEGF. RESULTS: The transfectants stably expressing antisense VEGF121 were observed to produce markedly reduced 3.3 kb VEGF mRNA and 45 KD, 41 KD, 32KD VEGF proteins. When xenografted s.c. into nude mice, growth and metastasis of the antisense-VEGF transfected cell lines were greatly inhibited when compared with control cells. CONCLUSION: Antisense VEGF gene significantly inhibited tumor growth and metastasis and may provide an experimental example for the development of antiangiogenic gene therapy. PMID- 11866938 TI - [Effects of antisense human telomerase RNA on growth of human gastric cancer cells]. AB - OBJECTIVE: To observe the inhibitory effects of antisense human telomerase RNA (hTR) on growth of human gastric cancer cells. METHODS: SGC7901 cell line was transfected with antisense hTR expression vector (pBBS-hTR) by lipofectAMINE. The gene transfected cells were cultured in vitro and then inoculated into nude mice. RESULTS: Antisense hTR expression was enhanced and sense hTR expression inhibited in clones containing antisense hTR after hygromycin resistant selection which induced tumor cells apoptosis. The growth rate of the major gene transfected cells in culture and tumorigenesis of the transfected cells in nude mice were significantly reduced in comparison with those of controls. The survival time of nude mice inoculated with the gene transfected cells was markedly prolonged when compared to controls. CONCLUSIONS: The malignant proliferation of gastric cancer cell can be reversed by antisense hTR induced cell apoptosis. Thus antisense hTR may serve as a potential therapeutic approach to the treatment of human cancer. PMID- 11866939 TI - [Expression of proliferation and apoptosis related genes in colorectal adenoma with malignant transformation]. AB - OBJECTIVE: The effect of proliferation and apoptosis related genes of colorectal adenoma with malignant transformation and its clinicopathologic significance was studied. METHODS: p53, bcl-2, Fas, bax expression were detected immunohistochemically in 61 cases of colorectal adenoma with either mild or severe dysplasia and carcinoma. The rate of proliferation was evaluated with Ki 67 and apoptosis with TUNEL. RESULTS: The positive rates for p53 and bcl-2 protein in carcinomatous area, adenoma with severe and mild dysplasia were 67.2%, 57.4%, 19.7% and 77.1%, 80.3%, 49.2% respectively. The mean Ki-67 index and PCD index were 38.4%, 36.5%, 16.1% and 2.2%, 1.9%, 0.5% respectively. The difference of the expression of p53 protein, bcl-2 protein and Ki-67 index, PCD index in carcinomatous region and adenoma with severe dysplasia compared with mild dysplasia was statistically significant (P < 0.05). CONCLUSION: It is apparent that both apoptosis inhibition and abnormal proliferation play an important role in tumor formation. Adenoma with severe dysplasia is a lesion very close to carcinoma in nature, which possesses active tendency to become fully malignant. Detection of these proteins is important in the early diagnosis of malignant transformation of an adenoma. PMID- 11866940 TI - [The prognostic significance of expression of c-met oncogene and its relation to gastric mucosal lesions]. AB - OBJECTIVE: To determine whether there is a relationship between the expression of c-met oncogene and the stage of gastric mucosal lesions or the prognosis of gastric carcinomas. METHODS: Expression of c-met was investigated in 169 gastric mucosal lesions by using immunohistochemistry. Survival analysis was studied with the Kaplan-Meier test. RESULTS: For chronic superficial gastritis, chronic atrophic gastritis, intestinal metaplasia, dysplasia, early gastric carcinomas and advanced gastric carcinomas, the expression rates were 23.5%, 36.8%, 51.5%, 61.3%, 66.7% and 73.7% respectively. The positive rate was higher in intestinal metaplasia, dysplasia and gastric carcinomas than in chronic superficial gastritis (P < 0.05). Expression of c-met increased with the increasing proliferation of gastric mucosa in rank correlation test, which showed a close relationship (P < 0.01). Expression of c-met correlated significantly with histologic type, serosal invasion and lymph node metastases. In particular, the expression of c-met was significantly higher in Borrmann type IV gastric carcinomas (87.5%) than in early gastric carcinomas (66.7%) or Borrmann type I, II (68.7%, P < 0.01). The survival rate of patients with expression of c-met was significantly lower than that of patients without c-met expression. CONCLUSION: The expression of c-met is associated with proliferation and malignant transformation of gastric mucosa. These results suggest that expression of c-met might be a new prognostic factor in gastric carcinoma. PMID- 11866941 TI - [The expression of DNA mismatch repair genes and detection of DNA ploidy in young patients with colorectal carcinoma]. AB - OBJECTIVE: To investigate the clinicopathologic characters and carcinogentic pathways of young (age < 36) colorectal carcinomas (CRCs) in Guangzhou, China. METHODS: Immunohistochemistry and flow cytometry methods were used to detect the expression of hMSH(2) and hMLH(1), status of DNA ploidy in 63 cases of young CRCs from Guangzhou, China, and analyze their correlations with patient's clinicopathological characters. RESULTS: Of the 63 young CRCs studied, forty-four (69.8%) tumors were non-mucinous carcinomas, thirty-nine (61.9%) patients were in Dukes' C or D stage. Of the 59 CRCs which were successfully detected by immunohistochemistry and flow cytometry, ten (16.9%) CRCs lost either hMSH(2) or hMLH(1) and showed DNA diploid or near-diploid, while twenty-six (44.1%) had aneuploid DNA content and all with the normal expression of hMSH(2) and hMLH(1). In addition, there existed a significant percentage (23/59, 39%) of young CRCs showing no loss of either of these two mismatch repair proteins and having a diploid or near diploid DNA content. CONCLUSION: The overall percentage of young CRCs in Guangzhou is significantly higher than those in Caucasian predominant countries and about seventy percent of young CRCs in Guangzhou are conventional carcinomas. 39% of young CRCs in Guangzhou showed no evidence of either chromosomal instability or microsatellite instability carcinogentic pathway, indicating that there must be at least a third pathway which triggers the CRCs in these special subgroups of young patients in Guangzhou, China. PMID- 11866942 TI - [Cushing syndrome--a pathologic analysis based on 216 resected specimens]. AB - OBJECTIVE: To study the morphological features of adrenal cortex in Cushing's syndrome and the correlation of morphological changes with clinical manifestations. METHODS: 216 cases of Cushing's syndrome were collected. Among these there were 160 cases of adrenal hyperplasia, 50 cases of adenoma and 6 cases of adenocarcinoma of adrenal cortex. The morphometric method was used to measure the thickness of adrenal cortex microscopically. The weight of the resected specimens were carefully recorded and correlated with clinical laboratory findings and clinical manifestations. RESULTS: In adrenal cortex hyperplasia, the thickness and weight of the adrenal cortex increased gradually in the order of mild, moderate and severe hyperplasia. The hyperplasia and hypertrophy of the compact cell are very common in these specimens. The frequency of compact cell predominant cases was also increased in severe hyperplasia. The weight and size of the tumor differed significantly between adenoma and adenocarcinoma of the adrenal cortex. CONCLUSION: In Cushing's syndrome, adrenal cortex hyperplasia is the most common. The grade of hyperplasia was well correlated with clinical laboratory findings. Increase of frequency of compact cell predominance is important morphological feature of hyperplasia of adrenal cortex. Adenoma should be considered when atrophy of adjacent cortex is present. The tumor size and weight as well as mitosis are important parameters in differential diagnosis between adenoma and adenocarcinoma of the adrenal cortex, but the infiltration of capsule and blood vessels and/or metastasis still remain the gold criteria for diagnosis of adrenal adenocarcinoma. PMID- 11866943 TI - [An etiological and pathologic study of Rhinoscleroma]. AB - OBJECTIVE: To investigate the clinicopathologic characteristics and mechanism of tissue injury caused by Klebsiella rhinoscleromatis (KR) infection. METHODS: Thirty-three cases of rhinoscleromas in the nasal and pharyngeal region seen in this hospital from Jan. 1980 to Mar. 2000 were studied retrospectively. Warthin Starry (W-S) stain was used to study all cases. Five cases were studied further with transmission electron microscope. RESULTS: W-S stain revealed a great number of KR in Mikulicz cells within the granuloma. Electron microscopy demonstrated a number of phagosomes in the cytoplasm of Mikulicz cells where many KR were found. A small number of other organelles such as endoplasmic reticulums and lysozymes were squeezed to the side of cells. There were many granular substances on the surface of intracellular bacteria, which were not found on the extracellular KR. CONCLUSION: KR is a facultative intracellular bacteria that is able to resist the digestion of macrophages and to proliferate in them. Formation of granulomas and fibrosis in KR infiltrated regions is the major cause of tissue injury. PMID- 11866944 TI - [Pathological study of the pleomorphic carcinoma of the lung]. AB - OBJECTIVE: A clinicopathological study of 10 patients with pleomorphic carcinoma of the lung. METHODS: Histopathological and immunohistochemical staining for keratin, vimentin, Mac387, desmin, actin and S-100 protein were used for this study. RESULTS: Pleomorphic carcinoma of the lung was found to often occur in males above 50 years of age and with clinical symptoms including cough, expectoration, haemoptysis and chest pain. The most frequent microscopic diagnosis was squamous cell carcinoma, and adenocarcinoma, accompanied by spindle and giant cells. The epithelial component of pleomorphic carcinoma of the lung displayed positivity for keratin and the spindle cells displayed positivity for vimentin. In some cases the neoplastic epithelial component and spindle cells showed positive expression of both keratin and vimentin. CONCLUSION: Pleomorphic carcinoma of the lung may display various histopathological changes making it easy to be misdiagnosed as carcinosarcoma. Understanding its pathogenesis and histopathology is important for the diagnosis and differential diagnosis. PMID- 11866945 TI - [Pathological features of liver biopsy from 66 cases of long term blood donors with chronic hepatitis C]. AB - OBJECTIVE: To study the pathological features of liver biopsy from long-term blood donors with chronic hepatitis C. METHODS: From 250 long-term blood donors with positive serum anti-HCV antibody for more than five years, 66 donors with abnormal ALT level were selected and enrolled in this study. Liver biopsy was performed on these cases and the specimens stained with HE, and reticulin. Modified Schemer's classification for grading and staging was used during pathological observation. Twenty biopsies with serum HCV-RNA levels higher than 5.0 Meq/ml (bDNA) were studied immunohistochemically (SABC) for NS5. RESULTS: All cases displayed mild pathological changes. Grade 1 (G(1)) inflammation was 4.8% (3/63), G(2) 63.5% (40/63), G(3) 28.6%(18/63), G(4) two case. Stage 1 (S(1)) fibrosis was 60.3% (38/63), S(2) 34.9% (22/63), S(3) 3.2% (2/63), S(4) only one case. The main lesion was periportal inflammation, such as bile duct damage (100.0%), dense lymphoid aggregates (82.5%) and piecemeal necrosis (89.5%). Intralobular injury of hepatocytes was rather mild. Bridging necrosis and steatosis were not common and observed in only 12.3% (8/66) and 9.2% (6/66) respectively. As for fibrosis, proliferation and deposition of collagen were mainly distributed in the portal and perisinusoidal areas. Among 20 specimens with serum HCV-RNA levels higher than 5.0 Meq/ml (bDNA), all bile duct epithelial cells were positive for HCV-NS5 antigen, while hepatocytes in 4 cases were negative. CONCLUSION: (1) The pathological changes in this series show overall similarity with that of patients with posttransfusion hepatitis C. The main damages are portal and periportal inflammation, such as lymphoid aggregates, bile duct damage, piecemeal necrosis, and extensive periportal and perisinusoidal fibrosis. Intralobular damage was mild with occasional steatosis, which may be related to malnutrition, low level of serum iron due to long term donation. (2) Epithelial cells of the bile duct may be of another site for virus replication, thus it is related with bile duct damage and periportal inflammation. PMID- 11866946 TI - [Endometrial stromal sarcoma with multi-differentiation: a study of 17 cases]. AB - OBJECTIVE: To investigate the clinical and pathomorphological features of multi differentiated endometrial stromal sarcoma of the uterus and to discuss their behaviour and differential diagnosis. METHODS: The histological characteristics of all cases were observed by pathological examination, some of them have been studied by immunohistochemical and/or ultrastructural techniques. RESULTS: Multi differentiation was present in 13 cases of low grade and 4 cases of high grade endometrial stromal sarcoma, of which, 13 cases had sex-cord differentiation, 10 cases had smooth muscle differentiation, osseous differentiation in 2 cases and striated muscle differentiation in 1 case. Two types of multi-differentiation was present in 9 cases. CONCLUSIONS: Both low-grade and high-grade endometrial stromal sarcoma of uterus can display multi-differentiation. Sex-cord and smooth muscle differentiation are the most common types. Osseous and striated muscle differentiation are very rare. There is no definite correlation between prognosis and the amount or types of multi-differentiation components. PMID- 11866947 TI - [Cerulenin inhibits growth of human colonic carcinoma in nude mice]. AB - OBJECTIVE: To determine the effects of fatty acid synthase inhibitor, cerulenin, on tumor growth of human colonic carcinoma (LoVo) in nude mice. METHODS: The transplantable colonic carcinoma cell line LoVo model was established in BALB/CA nu/nu mice by subcutaneous implantation. Cerulenin was administrated by i.p. injection (80 mg/kg, 160 mg/kg each time) for 10 times. 17 days later the mice were killed and tumor size evaluated. RESULTS: The size of tumors in cerulenin treated group were significantly smaller than in control group and were similar to that of the 5-Fu treated group. Transmission electronic microscopy (TEM) revealed that death of LoVo cells treated by cerulenin was associated with apoptosis characterized by cell shrinkage, condensation and chromosome fragmentation. bcl-2 oncoprotein expression rate was significantly lower in cerulenin-treated group in comparing with the control group. The expression rate of bax was the opposite to that of bcl-2. CONCLUSIONS: Cerulenin effectively suppresses the growth of human colonic carcinoma cells in vivo. The mechanism of cell death may be correlated with apoptosis, and bcl-2 and bax gene may play an important role in regulating cerulenin-induced apoptosis. PMID- 11866948 TI - [Effects of hypoxia alone or exercise combined on capillarization of rat gastrocnemius muscle and its mechanism]. AB - OBJECTIVE: To study the effects of hypoxia and hypoxia-combined-exercise on capillary density and vascular endothelial growth factor (VEGF) and its receptor KDR of skeletal muscle in rats. METHODS: Myosin-ATPase histochemistry was used to assay the size and capillary density of skeletal muscle. VEGF and its receptor KDR were studied by immunohistochemistry. RESULTS: Five-week hypoxia (simulated 5,000 m altitude) resulted in a decrease in cross-sectional area of skeletal muscle fiber and an increase in capillary density (CD), but the capillary/fiber ratio (C/F) remained unchanged. After 5-week-exercise at high altitude (1 h/d, 6 d/w), the muscle fibers did not undergo atrophy. At the same time, CD and C/F were increased. VEGF protein was found primarily in the matrix between muscle fibers; VEGF receptor-KDR was shown mainly in endothelial cells of capillary. VEGF was more strongly stained in the skeletal muscle of hypoxia-combined exercise rats than the other two groups. CONCLUSION: Hypoxia itself can not induce neovascularization, while hypoxia-combined-exercise rats show capillary proliferation in skeletal muscle. VEGF and its receptor might play roles in this process. PMID- 11866949 TI - [Myoid/myofibroblastic differentiation in dermatofibrosarcoma protuberans: a clinicopathologic study of six cases]. AB - OBJECTIVE: The nature of myoid/myofibroblastic differentiation in dermatofibrosarcoma protuberans (DFSP) and its clinical and pathological significance were studied. METHODS: 124 DFSPs were reviewed by light microscopy. 6 cases with areas of myoid/myofibroblastic differentiation were assayed with immunohistochemical technique and electron microscopy was applied in two cases. RESULTS: Myoid/myofibroblastic differentiation occurred most commonly in fibrosarcomatous DFSP (FS-DFSP). It was recognized histologically as peripherally distributed or randomly scattered small eosinophilic nodules or short bundles, which were composed of bland spindle cells, closely resembling smooth muscle cells or myofibroblasts. Immunohistochemically, cells in myoid/myofibroblastic areas showed positive staining for alpha-SMA, MSA and vimentin, but negative for desmin and CD34. Electron microscopic study displayed the presence of microfilament bundles, focal dense bodies and micropinocytic vesicles, consistent with those of myofibroblasts. CONCLUSION: Myoid/myofibroblastic areas in DFSP possibly represents the hyperplasia of stromal myofibroblasts, rather than true myofibroblastic differentiation of the neoplastic cells. PMID- 11866950 TI - [Detection of methylation status of p16 tumor suppressor gene in soft tissue leiomyosarcoma]. AB - OBJECTIVE: To observe the methylation status of p16 tumor suppressor gene in human soft tissue leiomyosarcoma (LMS) and its relationship with p16 protein expression. METHODS: Methylation-specific PCR (MSP) assay was used to analyze the methylation status of 5' CpG island promoter/exon I region of p16 tumor suppressor gene in 38 LMS, 10 leiomyomas and 5 normal smooth muscle tissues. p16 expression in these tissues were examined by immunohistochemical assay. RESULTS: Hypermethylation of p16 was observed in 23.7% (9/38) of cases, of which 7 with negative pl6 staining and 2 with faint positive staining. In the cases without protein expression, hypermethylation rate of p16 was 50% (7/14). CONCLUSIONS: Epigenetic change due to 5' CpG methylation is the main cause of inactivation of p16 (INK4) gene which may be involved in tumor pathogenesis. PMID- 11866951 TI - [Studies of the clinicopathological changes of eight patients with lipid storage myopathy]. AB - OBJECTIVE: The clinicopathological analysis of eight patients with lipid storage myopathy are presented. The pathogeny and therapeutic effect are probed into. METHODS: Eight cases of lipid storage myopathy diagnosed by muscle biopsies with microscopic and electron-microscopic examination are analyzed. Quadriceps or biceps were biopsied. Muscle samples were stained with routine histology and histochemical enzyme and inspected by microscopy. Thin sections were stained with uranyl acetate followed by lead citrate prior to examination in a electron microscopy. Also, the therapeutic drugs of eight patients were evaluated. RESULTS: Vacuole or crack of muscular fibers involved all eight patients. Sudan Black B and Oil Red O stains demonstrated increase of lipid droplets within muscle fibers. Ultrastructural examination revealed numerous lipid droplets dispersed throughout the residual myofilaments. Three cases with pathologic changed muscular fibers occupying less than 1/5 were belong to low-grade, two cases (between 1/5 to 1/3) were moderate, three cases (more than 1/2) were severe. There was one case accompanying glycogen storage disease. One case was concomitant with deficiency of cytochrome C oxidase. After prednisone treatment, seven cases had greatly improved and one case failed to respond to. Treatment using vitamin B(2) together with other vitamins brought about a striking effect. Carnitine was very effective on the patients with system deficiency of carnitine. CONCLUSIONS: The pathogeny of lipid storage myopathy is varied. The confirmed diagnosis is depend on pathological features of muscle biopsy. Treatment with prednisone, carnitine, vitamins and food containing carnitine rich is very effective. It should be select the special treatment method if the pathogeny is clear. PMID- 11866952 TI - [A quantitative pathological study on angiogenesis, vascular endothelial growth factor and inducible nitric oxide synthase in astrocytomas]. AB - OBJECTIVE: To investigate whether the expression of vascular endothelial growth factor (VEGF) and inducible nitric oxide synthase (iNOS) are associated with angiogenesis and malignancy of astrocytomas. METHODS: Immunohistochemistry, semiquantitation and image analysis were used to study the expression features of VEGF and iNOS in 61 cases of astrocytomas having follow-up data. Factor VIII related antigen (FVIII RAg) was also analysed in reflecting the epithelial cell amount and the microvessel density. RESULTS: Microvessels in astrocytomas could be divided into seven types. The positive reaction areas and IA values of FVIIIRAg expression in the endothelium were significantly increased with the increase of tumor grade (P < 0.001) and VEGF labeling index (LI) (P < 0.05). The patients with high VEGF LIs (LI >or= 25%) had a poor prognosis. VEGF expression was significantly related to that of iNOS (P < 0.001), i.e., VEGF level was reduced along with the decrease of iNOS reaction. CONCLUSION: Detection of FVIIIRAg could reflect the angiogenesis activities in astrocytomas. VEGF and iNOS could up-regulate each other to promote angiogenesis and are of importance in determining the malignancy degree of astrocytomas. PMID- 11866953 TI - [E-cadherin associated protein expression and its significance in invasive lobular carcinoma and invasive ductal carcinoma of breast]. AB - OBJECTIVE: To study E-cadherin associated proteins alpha-, beta-, gamma-catenin expression and their significance in invasive lobular carcinoma (ILC) and invasive ductal carcinoma (IDC) of breast. METHODS: Immunohistochemistry staining (labeled streptavidin-biotin method) was used to detect alpha-, beta-, gamma catenin expression in invasive breast carcinoma (19 cases of ILC and 32 cases of IDC ). RESULTS: The loss and reduction rate of alpha-, beta-, gamma-catenin expression in ILC was 78.9% (15 cases), 52.6% (10 cases) and 84.2% (16 cases), respectively. The rate of alpha-, beta-, gamma-catenin expression showing loss and reduction in IDC was 75.0% (24 cases), 43.8% (14 cases) and 81.3% (26 cases), respectively. The staining intensity of alpha-, beta-, gamma-catenin expression in invasive carcinoma of breast was weaker than that in foci of carcinoma in situ (semiquantitative grading). There was a positive relationship between the expression of alpha-catenin and beta-catenin in invasive breast carcinoma. No significant association was seen between reduced protein expression of alpha-, beta-, gamma-catenin and the development of lymph node metastasis. CONCLUSION: alpha-, beta-, gamma-catenin expression was almost identical in ILC and IDC of breast, showing significant loss and reduction of protein expression in the carcinoma cells of invasive breast carcinoma, suggesting that alpha-, beta-, gamma-catenin may lose their normal adherent ability in the pathogenesis of invasive breast carcinoma. PMID- 11866954 TI - [Detection of Immunoglobulin light chain restriction of mucosa-associated lymphoid tissue type gastric lymphoma using mRNA in situ hybridization]. AB - OBJECTIVE: To evaluate the usefulness of in situ hybridization for Immunoglobulin kappa and lambda light chain mRNA to detect light chain restriction in the diagnosis of primary gastric lymphoma (PGL) of mucosa-associated lymphoid tissue (MALT) type. METHODS: Twenty-seven cases of PGL of MALT type and 5 cases of gastritis were analyzed using in situ hybridization with fluorescent isothiocyanate (FITC)-labeled oligonucleotide probes for kappa and lambda light chain mRNA. The kappa:lambda ratios of tumor cells, lymphocytes and plasma cells were analyzed to detect light chain restriction and clonal plasma cells. RESULTS: Light chain restriction was found in 10 of 27 (37%) cases of PGL cases (in 5/9 low grade and 5/18 in high grade). Clonal plasma cells were detected in low grade but not in high-grade cases. No light chain restriction was found in gastritis specimens, the adjacent tissue of tumors or the distant stomach mucosa in lymphoma cases. CONCLUSIONS: The detection of clonality using mRNA in situ hybridization could be a helpful diagnostic tool for gastric lymphoma. The presence of clonal plasma cells can be a marker of the early lesion of low-grade PGL. PMID- 11866955 TI - [Mutations of APC and K-ras gene in aberrant crypt foci from human colon]. AB - OBJECTIVE: To analyze the mutations of K-ras and APC gene in normal colorectal mucosa, aberrant crypt foci (ACF), adenoma and colorectal carcinoma (CRC); To study the genetic alteration in ACF and its possibility of being a molecular marker of very early colon cancer and to explore the relationship of ACF and colorectal adenoma. METHODS: DNA from 35 CRC, 15 adenomas, 34 ACF and 15 cases of normal mucosa with mucous glands were isolated by micro-dissection. Direct gene sequencing of k-ras gene including codon 12, 13 and 61 as well as the mutation cluster region (MCR) of APC gene. RESULTS: Mutation frequency of k-ras gene in ACF, adenoma and carcinoma was 17.6% (6/34), 13.3% (2/15), and 14.3% (5/35) respectively, showing no difference between the three pathologic changes. The ras gene mutation sites in adenomas, carcinomas and 4 ACF were limited to codon 12 (GGT-->GAT), but in 2 ACF, they were located in codon 13 (GGC-->GAC). K-ras gene mutation in ACF was found more frequently in old patients and in patients with polypoid cancers. No mutations were found in codon 61 in three types of tissue. Mutation rate of APC genes in adenomas and carcinomas was 22.9% (8/35) and 26.7% (4/15) respectively, which was higher than that of ACF 2.9%, (P < 0.05). APC gene mutation in carcinomas was not correlated with age, location, size and differentiation. CONCLUSION: The morphological changes and gene mutation status are different in ACF and adenomas. ACF is a very early morphological lesion in the carcinogenesis of colorectal carcinoma. Therefore, ACF is considered to be the putative "microadenoma" where the development of colorectal carcinomas may be from "normal epithelium to ACF and then to carcinomas". PMID- 11866957 TI - [Clinical pathological analysis of adenomatoid tumor in uterus and ovaries]. AB - OBJECTIVE: To study the immunohistochemical feature and the differential diagnosis of adenomatoid tumors in uterus and ovaries. METHODS: Clinical pathological analysis and immunohistochemical studies were performed on 24 cases of adenomatoid tumors in the uterus and ovaries. RESULTS: Of the 24 cases, 21 cases were in the uterus, 2 cases in the ovaries and 1 cases in both the uterus and the ovary. Grossly, the mean diameter of the 22 uterus tumors was 2.2 cm, ranging from 0.2 - 5.5 cm. 14 (63.6%) were located in the subserosa or near by the subserosa of the uterine cornua. The other 8 tumors located in the myometrium. The cut surface presented a nodular pattern with grayish white or yellowish in color, partially cystic. 3 ovarian tumors became all cystic, without a clear-cut margin from the surroundings. Microscopically, the tumor consisted of various gland-like structure or luminal spaces lined with flat, cuboidal or low columnar cells, similar to blood vessels in structure. Among the tumor cells, there were scattered vesicular cells with large or small vacuoles, but no nuclear atypia and mitotic figures detected. Immunohistochemical staining showed the tumor cells positive for vimentin, AE(1)/AE(3) and calretinin, but negative for F VIII-Rag. S-100 and EMA were positive in 20 (83.3%) and 4 (16.7%) cases respectively. CONCLUSION: Adenomatoid tumor of the female genital tract is mesothelial in origin and uterus was considered as the most common site of occurance. Immunohistochemical phenotypes can be used as an important evidence for differential diagnosis. The biological behavior of adenomatoid tumor is benign and with a good prognosis. PMID- 11866956 TI - [DNA ploidy analysis and Ki-67 examination on renal neoplasms: investigation of degree of malignancy and biological behavior of small renal tumors]. AB - OBJECTIVE: To investigate the degree of malignancy and the biological behavior of small renal tumors and provide a pathological basis for clinicians in determination of further treatment and prognosis. METHODS: Flow cytometry and immunohistochemical studies were performed on 48 small tumors with diameters less than 3 cm. and 39 renal cell carcinomas with diameters larger than 3 cm in comparison to assess their DNA ploidy status and Ki-67 PR (proliferation rate). RESULTS: Of the 48 small renal tumors, 6 adenoma were all diploid with low PR (< 1%). Aneuploid rate of 42 small renal tumors with clear and granular cell type was 16.7%, showing no statistical difference with the 33.3% aneuploid rate of 39 large renal cell carcinomas. 42 small renal tumors had a lower mean PR than large renal cell carcinoma. DNA ploidy correlated closely with nuclear grade, stage and histological type. There existed a close relation between PR and grade. CONCLUSION: Small renal tumors consist of both adenoma and carcinoma, therefore tumor size can not be used as a criterion in their differential diagnosis. DNA ploidy and Ki-67 PR are important indicators for predicting the biological behavior and prognosis of renal parenchymal tumors. PMID- 11866959 TI - [Gene rearrangement and p53 expression in defining the nature of angioimmunoblastic lymphadenopathy]. AB - OBJECTIVE: To investigate gene rearrangement and p53 expression in defining the nature of angioimmunoblastic lymphadenopathy. METHODS: DNA was extracted from paraffin-embedded tissue blocks of 44 angioimmunoblastic lymphadenopathy (AIL) patients and analyzed with polymerase chain reaction (PCR) for IgH and TCRgamma gene rearrangement. Immunohistochemistry staining was used to detect p53 protein expression. Thirty-five cases were followed-up. RESULTS: 12 out of 44 cases (27.3%) showed TCRgamma gene rearrangement and 2 (4.5%) showed IgH gene rearrangement. Rearrangement of both IgH and TCRgamma genes were detected in 2 cases (4.5%). 14 cases (31.8%) showed p53 positive expression, among which 12 showed positive rearrangement and 2 showed negative (P < 0.01). Eight out of 11 patients of positive gene rearrangement died in one year, while only 3 patients were still alive at the eighteenth month of follow-up, three of 24 patients of negative gene rearrangement were found dead at the time of the one year follow up, while the rest 21 patients were alive and the longest survival time was 96 months. CONCLUSIONS: Gene rearrangement can define the pathological nature of AIL. The expression of p53 is highly related to gene rearrangement, and thus an important immunological marker in research on AIL. PMID- 11866958 TI - [Pathogen detection in the myocardial lesion using in situ RT-PCR in mice induced by experimental CoxB(3m) virus infection]. AB - OBJECTIVE: To detect the location of pathogens in myocardium using in situ RT-PCR technique in order to study the pathogenetic course of the myocardial lesion induced by CoxB(3m) virus infection in mice. METHODS: (1) Thirty and fifty Balb/c mice were used respectively to establish the acute and chronic CoxB(3m) infected models, with another 25 healthy mice as the controls; (2) KS400 image analysis system (Germany) was used to measure the cardiac chamber area and the left ventricular wall thickness of the chronic infected mice and the controls; (3) CoxB(3m) virus in myocardial tissue was detected using in situ RT-PCR by direct incorporated technique which employed nucleotide labeling by anti-digoxin antibody and bonded with alkaline phosphatase (anti-dig-AKP method). RESULTS: Picture analysis indicated that the left ventricular chamber area was enlarged and the left ventricular wall was thinner in the chronic repeated virus infected models than those of the controls. With in situ RT-PCR, positive signals for Coxsackie virus B(3m) RNA were detected not only in the myocardium of the acute Balb/c mice models but also in the myocardium of the chronic mice models. CONCLUSION: Coxsackie virus B(3m) is able to induce pathologic lesions by exhibiting positive CVB-RNA signals in both acute and chronic models in mice. In the chronic experimental models, the cardiac chamber is enlarged while the ventricular wall is thinned which demonstrates the association with persistent infection of Coxsackie virus B(3m) virus. PMID- 11866960 TI - [Mycobacterial spindle cell pseudotumor of lymph nodes]. AB - OBJECTIVE: To study the clinicopathological of mycobacterial spindle cell pseudotumor in lymph nodes. METHODS: Two cases of spindle cell pseudotumor in the lymph nodes of infants were examined using light microscopy. The sections were stained with HE, Ziehl-Neelsen and studied immunohistochemically. RESULTS: Both infants were 1 year of age and had a history of Bacilli Calmette Guerin (BCG) vaccination before discovery of their lymph node lesions. Clinically, the patients presented fever and enlarged lymph nodes. Histopathologically, the lesions showed proliferation of spindle cells arranged in bundles, storiform patterns with abundant capillaries and lymphocyte, plasma cell and neutrophil infiltration forming a spindle cell pseudotumor. Most of the spindle cells were phagocytic cells which contained large amounts of mycobactrium as observed by Ziehl-Neelsen staining. Immunohistochemical staining showed strongly positive result for Mac387 and lysosome, but negative for S-100 in spindle cells and histocytes. CONCLUSIONS: Mycobacterial spindle cell pseudotumor of lymph nodes may occur not only in immunodeficient patients (such as AIDS) but also in infants after BCG vaccination. The lesions may mimic spindle cell tumor in lymph nodes, therefore it is very important to recognize these lesions in order to treat the patient properly. PMID- 11866961 TI - [The relationship of clinicopathology and immunophenotypes in non-Hodgkin's lymphomas of the spleen]. AB - OBJECTIVE: To investigate the relationship between clinicopathologic features and immunophenotypes in non-Hodgkin's lymphoma (NHL) of the spleen. METHODS: Clinicopathologic analysis and follow up; immunohistologic staining by SP method and EBER in situ hybridization. RESULTS: (1) Splenomegaly was found in all 19 cases of spleen NHL, 10 of the 19 (52.6%) cases had masses or a mass in their enlarged spleen; (2) 14 of the 19 (73.7%) cases were of B-cell type (including 5 cases of follicular type and 9 cases of diffuse type), in which 8 cases were centroblastic, 3 of centroblast/centrocyte and 4 cases of small lymphocyte types; all 10 cases of primary splenic lymphomas (PSL) were B-cell type; (3) 5 of the 19 cases (26.3%) had peripheral T-cell origin, including 4 cases of large cell type and 1 case of small cell type; of the 3 TIA-1 positive cases, 2 were also positive for EBER 1/2 (one CD 8+ and another CD56+), the remaining case was CD8-, CD56-, EBER1/2-; all 5 cases were secondary NHL of the spleen; (4) Follow up data were obtained for 14/19 cases (73.7%). 8 of 9 surviving cases were primary NHL. The range of survival period varied from 8 to 10 years. All 5 deceased cases had secondary NHL and their survival period was 2 to 6 months. CONCLUSIONS: Some relationship exists between the clinicopathology and origin of the tumor cells in splenic NHL, the prognosis of primary NHL is better than that of the secondary NHL and the diagnosis of primary NHL should be made with discretion. PMID- 11866962 TI - [Primary cutaneous CD30-positive anaplastic large cell lymphoma]. AB - OBJECTIVE: To analyse the clinical manifestations, pathological and immunohistochemical features of 9 cases of primary cutaneous CD30-positive anaplastic large cell lymphoma (ALCL) in order to improve the criteria for its early and differential diagnosis. METHODS: Histopathological method was used to study the morphological characteristics. Immunohistochemical stainings (SP or ABC method) for leukocyte common antigen (LCA), CD20, CD30, CD45RO, CD68, epithelium membrane antigen (EMA), cytokeratin (CK) and HMB45 were applied to the routine paraffin sections of all the 9 cases. RESULTS: The 9 cases of primary cutaneous CD30-positive ALCL consisting of 6 males and 3 females, aged 31 to 84 years (mean 58.2 years). All patients presented with subcutaneous masses or papular eruptions on lower trunk and extremities. Histopathologically, the lesions were composed of numerous large round, oval or pleomorphic cells with abundant cytoplasm, amphophilic or basophilic, and large nucleus with distinct nucleolus. Mitosis was often observed. Multinucleated giant cells and Reed-Sternberg cells may be seen. The neoplastic cells displayed positive antigen markers for CD30 in 9 cases, 6 positive for CD45RO and 3 cases negative for both CD45RO and CD20. Two patients died of metastasis, 2 patients experienced recurrence and 5 patients still well during follow-up. CONCLUSIONS: Primary cutaneous CD30-positive ALCL is a morphologically distinctive neoplasm with a comparatively favorable prognosis. This tumor can be differentiated from other malignant tumors on the basis of histopathologic features and positive CD30 which is of significance. PMID- 11866963 TI - [Cytotoxic natural killer/T-cell lymphomas of the lymph nodes]. AB - OBJECTIVE: To investigate the clinicopathologic features of cytotoxic natural killer (NK)/T-cell lymphomas of the lymph nodes. METHODS: Clinicopathologic observations and follow up on 5 cases of cytotoxic NK/T-cell lymphomas of the lymph nodes, immunohistochemical staining for CD45RO, CD8, CD56, CD30, CD20, TIA 1 and in situ hybridization for EBER1/2 were performed. RESULTS: (1) The clinicopathologic features of cytotoxic NK-T-cell lymphomas of the lymph nodes were summarized; (2) The tumor cells were positive for CD45RO in 4 of the 5 cases, in which 3 cases were also positive for CD56. One case was of null cell type. The tumor cells were positive for TIA-1 and EBER1/2 in all 5 cases. CONCLUSIONS: The clinicopathological features and special immunophenotypes of cytotoxic NK/T-cell lymphomas of the lymph nodes were present in all 5 cases. Clinically, this entity tends to be an aggressive process with poor prognosis. PMID- 11866964 TI - [Clinicopathological significance of grading on thymic epithelial tumors]. AB - OBJECTIVE: To study the clinicopathologic relevance of a thymic epithelial tumor (TET) grading standard with the WHO classification. METHODS: A grading system for TET was proposed based on the application of WHO histological typing of thymic tumors and analyzed in relation to clinical therapy results and follow-up data of 200 TET cases. RESULTS: In this series, 8 patients (4.0%) belonged to type A, 68 (34.0%) were type AB, 17 (8.5%) were type B1, 39 (19.5%) were type B2, 27 (13.5%) were type B3 and 36 (18.0%) were type C. The remaining 5 cases were rare thymomas. The overall postoperative survival data showed highly significant differences among the histological subtypes (P < 0.001). Type A & type AB thymomas showed excellent prognosis, none of these patients died of tumor; in type B1, only 1 case (5.9%) died at 22 months postoperatively. Types B2, B3 and C thymomas shared the bad, worse and worst prognosis. Ninety-six patients (48.0%) were in stage I, 26 (13.0%) in stage II, 65 (32.5%) in stage III and 13 (6.5%) in stage IV. Clinical stage is also highly significant in predicting survival (P < 0.001). It was found that tumor histology could predict survival expectancies well in stage I and stage II cases. It was also found that type B2, B3 and C thymomas had a statistically significant worse prognosis than type A, AB and B1 thymomas (P < 0.001). According to the histology, clinical data, biological behavior and prognosis, it is proposed that thymomas be divided into 4 grades: grade I, II, III and IV. Follow-up is the best strategy for grade I & II patients after radical surgery. In this series, the 30 patients (15.0%) presenting clinical signs of myasthenia gravis were mostly in type B2 and B3 groups (P < 0.01). CONCLUSIONS: The WHO classification for TET provides good pathological definitions and criteria for diagnosis, which can independently predict the invasiveness and prognosis of TET. TET grading is of use in unifying pathological and clinical findings, in selection of proper therapy and in predicting prognosis. PMID- 11866965 TI - [Fine mapping of the loss of heterozygosity for chromosome 1pter-p36.11 in nasopharyngeal carcinoma]. AB - OBJECTIVE: To construct a detailed mapping of the Chromosome 1pter-p36.11 deleted region (63.4 cM) in nasopharyngeal carcinoma (NPC) by polymerase chain reaction loss of heterozygosity (PCR-LOH) analysis for the further research of NPC-related gene(s). METHODS: Biopsies from 47 cases of NPC patients were studied. DNA extracted from separated cancer cells and their corresponding non-cancer lymphocytes were amplified with PCR, followed by the analysis of LOH and microsatellite instability (MI) for 20 loci spanning Chromosome 1pter-p36.11 region with an average interval of 3.0 cM. RESULTS: 82.2% of NPC cases (37/47) showed at least one loci of LOH. The highest frequency of LOH was found at loci D1S234 on 1p36.13 (50.0%), with the LOH at loci D1S2644 on 1p36.22 slightly less (37.5%). The occurrence of LOH at D1S234 showed no significant difference for the cases at early stage and at advanced stage [60% (9/15) vs 50.0% (8/16) respectively, P > 0.05]. High frequencies of MI were detected at D1S243 on 1p36.33 (37.5%) and D1S199 on 1p36.21 (30.2%). CONCLUSIONS: There are two common deletion regions: one localized at 1p36.13 (D1S234, 2.0 cM) and the other at 1p36.22 (D1S436-D1S2644, 6.3 cM), with a MI loci at D1S199 between them. This suggests that one or more putative tumor suppressor gene(s) related to the early stage of NPC tumorigenesis may be encompassed in this zone. PMID- 11866966 TI - [Expression of cytokeratin 7 and 20 in ovarian metastatic carcinomas]. AB - OBJECTIVE: To study the distinctive clinicopathologic and immunohistochemical difference between ovarian metastatic carcinomas and primary ovarian carcinomas. METHODS: The clinical and pathological features of 27 cases of ovarian metastatic carcinomas (gastric carcinomas 12 cases, colon carcinomas 11 cases, others 4 cases) obtained from our department were reviewed. Immunostainings for CK (AE1/AE3), CK7, CK20, CEA, vimentin, nm23 were performed with SP staining methods. RESULTS: On gross examination, metastasis from gastric adenocarcinoma were usually bilateral, while solid (11/12) and metastases from colonic adenocarcinoma were more often unilateral and cystic (7/11). Microscopically, metastases from gastric adenocarcinoma revealed signet ring cells or poorly differentiated adenocarcinomas (12/12), whereas metastases from colonic adenocarcinomas showed similar morphology of endometrioid adenocarcinoma (8/11). The majority of ovarian metastases of gastric carcinoma (7/12) and colon carcinoma (8/11) were CK20 positive. In particular, CK20 was invariably expressed in colon cancer metastases. Most of the ovarian metastatic carcinomas from the gastrointestinal tract failed to react with immunostaining of CK7. A combined use of CEA, vimentin and nm23 had made a correct classification for 11/12 cases of the gastric carcinoma, 10/11 cases of the colonic cancer. CONCLUSIONS: CK7 and CK20 have been proved to be useful antibodies in distinguishing between metastatic carcinomas and primary carcinomas of the ovary. Combined use of a panel of antibodies can give more significant results. PMID- 11866967 TI - [Morphological observation of Mahaim's fibers in 7 cases]. AB - OBJECTIVE: To investigate the Mahaim's fibers of the cardiac conduction system (CCS) of human hearts and study their morphological features. METHODS: The CCS from 165 cases of sudden death without extracardiac causes and those from 760 cases of non-cardiac death were sampled using a method established by the authors. Serial sections were prepared for histological observation. RESULTS: Mahaim's fibers were discovered in 7 cases, 5 cases were in the sudden death group, accounting for 3% in that group, 2 cases were in the non-cardiac death group, accounting for 0.26% in that group. All cases belonged to the bundle ventricular branch type. The characteristics of Mahaim's fibers were found to be as follows. Their transverse diameter was 10 - 35 microns. The bodies of the Mahaim's fibers that originated from A-V bundle were slender with nucleus in the middle of the cell. While the shape of cells from the left bundle branch varied according to the origin of the Mahaim's fiber with size enlarging from up downward. Both types of cells showed abundant lightly stained cytoplasm, similar to A-V bundle and left bundle branch cells. The cells were not bifurcated and some of them were surrounded by a thin layer of fibrous tissue. CONCLUSION: Mahaim's fibers are not a rarity and it is an abnormal development of an A-V by path. PMID- 11866968 TI - [The effects of sodium butyrate on proliferation, differentiation and apoptosis in immortalized esophageal epithelial cells]. AB - OBJECTIVE: To study the effects of sodium butyrate on proliferation, differentiation and apoptosis of immortalized esophagus epithelial cells. METHODS: SHEE, an immortalized human fetal esophageal epithelial cell line induced by HPV18 E6E7, was cultivated in culture flasks and 24-well plates. Two experiment groups of cultured cells were treated with 1 and 5 mmol/L of sodium butyrate respectively for 4 days, and one group of untreated cells set aside as control. The numbers of cloned cells were calculated. The ultra-structure of SHEE cells was examined by transmission electron microscopy (TEM). The cell cycle and number of apoptotic cells were measured by flow cytometry, Ki-67 and cytokeratin of cells were detected by immunohistochemistry method and F-actin of cells labeled with phalloidin was examined by laser confocal scanning microscopy. RESULTS: Colony formations showed a significant decrease in the 2 experiment groups after 4 days of culture (P < 0.01). In the 1 mmol/L group, the cells at S phase were diminished and arrested at G(0)/G(1) phase. Compared with control group, Ki-67 positive cells were found decreased, while F-actin and cytokeratin were increased. Apoptotic cells in 5 mmol/L group were increased markedly. CONCLUSIONS: Sodium butyrate may induce SHEE cells growth arrest, differentiation and apoptosis. The effects depend on sodium butyrate concentration and time of exposure. Whether it can be used in combination with other anticancer drugs should be further studied. PMID- 11866969 TI - [Profile of telomerase and telomerase RNA expression in nasopharyngeal carcinogenesis of rats induced by N, N'dinitrosopiperazine (DNP)]. AB - OBJECTIVE: To investigate the profile of telomerase and telomerase RNA expression in nasopharyngeal carcinogenesis (NPC) induced by N, N'dinitrosopiperazine (DNP) and examined histolofically. METHODS: Nasopharyngeal carcinomas of rats were induced by DNP and examined histologically. PCR-ELISA and nested RT-PCR were used to assay telomerase and telomerase RNA expression at different stages in the nasopharyngeal tissues of rats. RESULTS: During the carcinogenesis process, telomerase activity increased along with the formation of a nasopharyngeal carcinoma. Telomerase expression was positively related with nasopharyngeal carcinogenesis. Telomerase RNA expression was present and did not change during the NPC process. Expression of telomease RNA was earlier than telomerase activation. Telomerase activation and telomerase RNA expressin were also detected in the pre-cancerous nasopharyngeal lesions. CONCLUSION: Telomerase activation may participate in the onset and progression of NPC, and is an early step in NPC. PMID- 11866970 TI - [The malignant potential of ovarian atypical endometriosis]. AB - OBJECTIVE: To investigate the malignant potential of ovarian atypical endometriosis and ovarian endometrioid adenocarcinoma. METHODS: 12 cases of ovarian endometroid adenocarcinoma with associated endometriosis were reviewed retrospectively. Microsatellite markers were used to detect the loss of heterozygosity on chromosome 10q23 in ovarian carcinoma and their coexisting endometriosis. RESULTS: Among the 12 cases, benign eosinophilic metaplasia was observed in 8 cases, 5 of which (41.7%) with atypia. In these 5 cases, a spectrum of benign-atypical endometriosis with a transition to endometrioid adenocarcinoma was identified, and common genetic lesions were also detected in 2 of the 5 atypical endometriosis and their coexisting carcinomas. CONCLUSION: Histological observations as well as the molecular genetic evidence indicate that ovarian atypical endometriosis may possess a precancerous potential or play a role in the pathogenesis of certain malignant ovarian cancers. PMID- 11866971 TI - [Ossifying fibromyxoid tumor of soft parts: a clinicopathological analysis of eight cases]. AB - OBJECTIVE: To study the morphological characteristics and immunophenotype of ossifying fibromyxoid tumor of soft parts (OFT) with a discussion of its histogenesis. METHODS: The clinical, pathological and immunohistochemical features of 8 cases of OFT were evaluated. RESULTS: All 8 cases were middle to old aged patients, ranged from 43 -- 78 years (mean 63 years). Clinically, the majority presented as slowly growing painless masses that located in subcutis of the proximal extremities. Histologically, the tumor was characterized by the following three unique features which have diagnostic value. (1) The tumor was well circumscribed and encapsulated with an incomplete bony shell composed of metaplastic bone within the capsule in most cases; (2) The tumor parenchyma consisted of lobules of variable size and cellularity. The tumor cells within each lobule were rounded to short spindled with pale-staining or eosinophilic cytoplasm; (3) The tumor cells arranged in nests, cords, or laciform pattern, and were embedded in a characteristic fibromyxoid to collagenized stroma. Immunohistochemically, all 7 cases tested were positive for vimentin and NSE, while 6 cases expressed S-100 protein and 2 cases expressed desmin. Follow-up information showed recurrences in two patients 2 and 15 years after local excision. CONCLUSIONS: OFT is a distinctive soft tissue tumor of potentially low grade malignancy which occurred predominantly in middle to old aged patients. The characteristic bony shell, the unique cytological appearance and arrangement of the cells are pathognomonic features of OFT. Our immunohistochemical result supported a Schwann-cell origin. PMID- 11866973 TI - [Clinical significance of c-myc oncogene amplification in primary hepatocellular carcinoma by interphase fluorescence in situ hybridization]. AB - OBJECTIVE: To investigate the relationships between c-myc oncogene amplification in primary hepatocellular carcinoma (HCC) and its clinical prognosis. METHODS: To detect c-myc oncogene amplification in 100 cases with primary HCC and in 6 cases with satellite nodes by interphase fluorescence in situ hybridization (FISH). RESULTS: c-myc amplification was detected in 92 of 100 (92%) cases with HCC, 37 of which (37%) showed high copy amplification (HC), 55 (55%) low copy amplification (LC) and 8 (8%) single copy or no amplification (SC). There wasn't statistically different significance between the frequency of c-myc amplification and clinical stage and tumor size, and pathological types (P > 0.05), however, in the 90 follow-up resected cases, the one-year rates of recurrence in HC, LC and SC were 70.6% (24/34), 39.6% (19/48) and 12.5% (1/8) respectively (P < 0.05) but the recurrence rates after more than one year were 14.70% (5/34), 14.6% (7/48) and 0 (0/8) respectively (P > 0.05). Meanwhile, two-year survival rate of patients with HC was significantly lower in comparison with those with LC and SC (P = 0.001). CONCLUSIONS: There is a high frequency of c-myc oncogene amplification in primary HCC and the fact that the patients with HC showed poorer prognosis indicates that the level of c-myc oncogene amplification may be a sensitive index in the prognosis of HCC. PMID- 11866972 TI - [A clinicopathologic study on 9 cases of epithelioid hemangioendothelioma]. AB - OBJECTIVE: To study the clinicopathological features of epithelioid (histiocytoid) hemangioendothelioma (EHE). METHODS: 9 cases of EHE were examined by light microscopy and immunohistochemistry, 1 case with ultrastructural observation. RESULTS: The patients' age ranged from 16 to 47 years (mean 32 years). The lesions occurred in both sexes and presented at a variety of sites including head and facial area (4 cases), upper limbs (3 cases) and lower limbs (2 cases). The morphological characters were: (1) "epithelioid" or "histiocytoid" tumor cells; (2) rounded or polygonal tumor cells arranged in nests, cords and irregular patterns; (3) the tumor cells were embedded in myxoid or hyaline matrix; (4) primitive lumen formation within a single tumor cell (intracytoplasmic lumens); (5) lack of pleomorphism, mitotic activity and necrosis; (6) presence of spindle cell hemangioendothelioma in some cases; (7) osteoclast like giant cells scattered throughout the entire lesion were observed in one case. Immunohistochemistry studies on 7 cases found that tumor cells were positive for vimentin (7/7), factor VIII-RAg (5/7), CD31 (5/7), CD34 (5/7), cytokeratin (2/7), CD68 and alpha-AT (1/7). Ultrastructural study of 1 case demonstrated Weibel-Palade bodies and an abundance of intermediate filaments in the cytoplasm. The follow-up of 6 cases found that 3 cases developed local recurrences but none of them metastasized. CONCLUSIONS: EHE is a distinctive low grade sarcoma whose etiology remains unknown. Recognizing its morphological characters is important to avoid confusion with other similar lesions. PMID- 11866974 TI - [Relationship between genetic alterations and clinicopathological features in intrahepatic cholangiocarcinoma]. AB - OBJECTIVE: To evaluate the pedigree of genetic alterations during the tumorigenesis of intrahepatic cholangiocarcinoma (ICC) and their correlation with clinicopathological features by analysis of loss of heterozygosity (LOH) in 6 tumor suppressor genes (APC, MCC, DCC, OGG1, p53 and RB1) and point mutations in Ki-ras-2 oncogene. METHODS: Genomic DNA was isolated from paraffin-embedded slides of 22 surgically resected ICC cases by microdissection-based PCR amplification and agarose gel electrophoresis. Genetic alterations were analyzed by direct DNA sequencing. RESULTS: The total frequency of alterations in 7 genes studied was 86.4% (19/22). Based on the pattern of altered genes and their correlation with clinicopathological parameters, the genetic alterations were classified into two groups: Group I (9/19, 47.4%): alterations in APC, MCC, DCC and Ki-ras-2,); Group II (10/19, 52.6%): alterations in p53, OGG1 and RB1. The average age of patients in Group I (mean age, 57.2 years) was significantly younger than those in Group II (mean age, 69.1 years) (P < 0.05). CONCLUSIONS: The occurrence and development of ICC was closely related with the accumulation and cooperation of multiple genetic alterations. The genetic alterations of APC, MCC, DCC and Ki-ras-2 may play crucial roles in the early stage of development of ICC, and the genetic alterations of p53, OGG1 and RB1 may play important roles in accelerating advanced progression of ICC. The detection of the pedigree of genetic alterations in ICC may provide useful information for evaluating the state of tumor progression and clinic prognosis. PMID- 11866975 TI - [The pathology and DNA quantitative study of renal clear cell carcinoma in children]. AB - OBJECTIVE: To investigate the pathological features of renal clear cell carcinoma (RCCC) in children and its relationship to the DNA content of cancer cells and DNA ploidy. METHODS: The pathologic morphology of 4 cases of RCCC in children were observed. Using image analysis instruments the quantity of tumor cell DNA was measured. RESULTS: The cytoplasm of the tumor cells were clear in all four cases. Papillary structures were present in these tumors, of which the papillary structure in two cases was over 50%, three cases had small calcified bodies with prominent bleeding and necrosis. Around tumor the glomerular and tubular of kidney always is normal the average DNA index was 1.31. They presented diploidy, high diploidy or subtetraploidy. CONCLUSIONS: The specific features of most RCCC in children are clear cytoplasm, most contain papillary structures, calcified bodies, often with bleeding and necrosis. Around tumor the glomerular and tubular of kidney is always normal. The DNA contents of cancer cells present diploidy, high diploidy or subtetraploidy. PMID- 11866976 TI - [A study of the correlation between p16 protein expression and DNA quantitative analysis in human gastric carcinoma cells]. AB - OBJECTIVE: To investigate p16 protein expression and its correlation with DNA quantity in gastric carcinoma (GC) cells. METHODS: p16 protein expression was quantitatively determined by flow cytometry (FCM) in 41 cases of GC and 10 cases of normal gastric tissues. DNA quantitative analysis of 41 cases of GC was performed by image cytometry (ICM). RESULTS: p16 protein expression in GC was significantly lower than that in normal gastric tissues (respective FI: 0.81 plus minus 0.09 and 1.00 plus minus 0.11, P < 0.01). The expression of p16 protein did not show correlation with such factors as sex and age in GC, but it was significantly stronger in well differentiated GC than that in poorly differentiated GC (P < 0.05). The mean of DNA relative ploid (U value) in GC was from 1.14 to 2.98. U value becomes higher with the increase in grading (P < 0.01). There was significant negative linear correlation between p16 protein expression and U value in GC (P < 0.01). CONCLUSION: Abnormal expression of p16 protein is closely correlated to the occurrence, development and malignancy degree of GC. The level of p16 protein expression implies important clinical significance for GC treatment and may serve as an indicator to predict the prognosis of patients. PMID- 11866977 TI - [Activation of p38 and c-Jun NH2-terminal kinase mitogen-activated protein kinases in human prostate carcinoma cell lines with different metastatic potentials]. AB - OBJECTIVE: To investigate the influence of P2 purinoceptor agonist ATP on p38 mitogen-activated protein kinases (MAPK) and JNK (c-Jun NH(2)-terminal kinase) signaling pathway in two human prostate cancer cell sublines (1E8 and 2B4) with different metastasis potentials. METHODS: Activated p38 and JNK were detected by Western blot with phospho-specific antibodies directed against the dually phosphorylated active forms of p38 or JNK. RESULTS: Exposure of 1E8 and 2B4 prostate cancer cells to ATP resulted in p38 activation in a concentration- and time-dependent manner. ATP-induced p38 activities were higher in metastatic 1E8 cells when compared with nonmetastatic 2B4 cells. JNK signaling pathway was not activated by ATP. An MEK inhibitor PD 98059 failed to inhibit ATP-induced p38 activation, while p38 inhibitor SB 203580 and P2 purinoceptor antagonist suramin effectively inhibited activation of p38, the inhibition rate being 1E8 83% and 79%, 2B4 81% and 69%. The response of ATP-stimulated p38 to G-protein modulator pertussis toxin in 1E8 and 2B4 cells was not obvious. CONCLUSIONS: p38 but not JNK signaling pathway can be activated by P2 purinoceptor agonist ATP. Differences of p38 activation by ATP are noted between metastatic 1E8 cells and non-metastatic 2B4 cells. These results provide instructive clues to cancer malignant growth and metastasis research. PMID- 11866978 TI - [Using Hsp70 promoter to regulate target gene expression in tumor]. AB - OBJECTIVE: To regulate gene expression in the desired tumor cells both in vitro and in vivo. METHODS: Heat inducible green fluorescent protein (GFP) expression plasmid and adenovirus were built by using different sizes of 5' end regulating sequence from human heat shock protein (Hsp) gene as promoter and GFP cDNA as report gene. GFP expression turned on by heating was observed in the cultured cells and the tumor grown in the dorsal skin window chamber. RESULTS: A 400 bp of Hsp gene 5' end regulating sequence can be activated by heating and it can drive report gene expression effectively both in vitro and in vivo. CONCLUSIONS: Heating can selectively induce gene expression in the targeted tumor. It provides an useful tool for cancer gene therapy because it possibly maximizes tumor cell killing and minimizes normal tissue damage. PMID- 11866979 TI - [Effects of hypoxia on the expression of protein kinase C mRNA in pulmonary artery smooth muscle and endothelial cell]. AB - OBJECTIVE: To investigate the effects of hypoxia on the expression of PKC mRNA in pulmonary artery smooth muscle cells (PASMC) and endothelial cells. METHODS: In situ hybridization technique was used to detect the expression of PKC mRNA in different segments of pulmonary artery smooth muscle and endothelial cell (EC) in rats, and the effect of hypoxia on the expression of PKC alpha mRNA in pulmonary artery smooth muscle and endothelial cells in vivo and in vitro. RESULTS: PKC alpha mRNA was expressed in normal PASMC and EC in rats. The expression of PKC alpha mRNA was higher in the PASMC of intra-acinar pulmonary arteriole than that of muscular arteriole (P < 0.01). In 14 and 28 days of hypoxia, the expression of PKC alpha mRNA was higher both in EC of intraacinar pulmonary arteriole and muscular arteriole and PASMC of intra-acinar pulmonary arteriole (P < 0.01). After 14 days of hypoxia, the expression of PKC alpha mRNA was not significantly changed. But by 28 days, it became significantly higher (P < 0.01). PKC alpha mRNA was expressed in normal PASMC and EC in pigs in vitro. After 1 h of hypoxia, no change was detected, but PKC alpha mRNA expression was significantly higher after 48 h and 72 h of hypoxia in PASMC and EC in vitro (P < 0.01). CONCLUSIONS: Hypoxia can promote expression of PKC alpha m RNA in PASMC and EC, especially in the PASMC of intra-acinar pulmonary arterioles. PKC alpha may play an important role in the development of hypoxia pulmonary hypertension. PMID- 11866980 TI - [Overexpression of bcl-2 protects HCC-9204 hepatoma cells from ethanol-induced apoptosis]. AB - OBJECTIVE: To explore the effect of apoptosis-associated gene bcl-2 overexpression on ethanol-induced apoptosis in hepatocellular carcinoma (HCC) cells. METHODS: The plasmid of retrovirus expression vector pDOR-SB, which contains human bcl-2 cDNA, was transfected into a HCC cell line HCC-9204 cells whose bcl-2 protein expression is below detectable level. The expression of bcl-2 protein was detected by immunohistochemical ABC method. The cells were cloned continually until a monoclonal cell strain which expressed bcl-2 protein at a 100% positive rate was obtained, and the cells were detected with flow cytometry. After the above cells were treated with 6% ethanol for 6 h, the happening of apoptosis were detected by MTT assay, TUNEL assay and flow cytometry. RESULTS: bcl-2 protein expression was detected in most of the cells which had been transfected with pDOR-SB, while it was bcl-2 protein negative in cells transfected with pDOR empty vector or non-transfected HCC-9204 cells. The obtained monoclonal cell strain expressed bcl-2 protein at a 100% positive rate under flow cytometer after being cloned 3 times in succession, and was named HCC bcl-2. After being treated with 6% ethanol, the optical absorbance data of HCC bcl-2 cells and non-transfected HCC-9204 cells detected by MTT assay were 0.519 +/- 0.053 and 0.366 +/- 0.046, respectively, the former was significantly higher than the latter (P < 0.01). Their TUNEL index was 0.387 and 0.613 respectively, and their sub-G1 apoptotic peak scale was 3.8% and 10.7% respectively, the former were all lower than the latter (P < 0.05). CONCLUSION: Overexpression of bcl-2 protein can suppress ethanol-induced apoptosis in HCC-9204 hepatoma cells. PMID- 11866981 TI - [Analysis of loss of heterozygosity on chromosome 6 in human prostate carcinoma and prostatic intraepithelial neoplasia]. AB - OBJECTIVE: To detect the significance of loss of heterozygosity (LOH) on chromosome 6 in prostate carcinoma and high grade prostatic intraepithelial neoplasia (PIN). METHODS: Pure DNA was obtained from prostate neoplasms and normal tissues after tissue microdissection. LOH of chromosome 6 was detected by PCR based microsatellite polymorphism analysis technique using 20 pairs of microsatellite primers in 10 prostate carcinoma cases and 10 high grade PIN cases. RESULTS: Allelic loss at one or more loci was observed on chromosome 6 in 8 of 10 prostate carcinoma cases. 6q21-6q23 and 6q25-6q27 were two of LOH high frequency regions. 5 high grade PIN cases had LOH detected on chromosome 6. CONCLUSIONS: Two high frequency LOH regions were detected on chromosome 6 of prostate carcinoma. Cyclin C, IGF2R genes were two candidate tumor suppressor genes located in these two regions, they may be involved in the initiation and progression of prostate cancer. PMID- 11866982 TI - [Pathomorphologic features of clinical stage A prostate carcinoma and causes for misdiagnosis and missed diagnosis]. AB - OBJECTIVE: To study the pathomorphologic features and causes for misdiagnosis and missed diagnosis of clinical stage A prostate carcinoma. METHODS: Prostate samples from a series of 1 020 prostate resections performed in five Shanghai hospitals were obtained, from which 50 clinical stage A carcinoma were studied by immunohistochemistry, according to tumor differentiation and their volume, they were divided into stage A1 and A2 disease. The pathomorphologic features of stage A1 and A2 disease were compared, and the causes for misdiagnosis were analysed. RESULTS: Stage A1 cancer were well- or moderately well-differentiated, of low volume and tended to be multifocal; Stage A2 cancers were poorly differentiated, of high volume, diffuse infiltration and with high grade prostatic intraepithelial neoplasia. 8 cases were misdiagnosed of which 7 cases were stage A1 and misdiagnosed as benign proliferous small gland lesions and 1 case of A2 disease was misdiagnosed as epithelioid histiocytic reactive proliferation. CONCLUSIONS: Stage A1 cancer tended to be initiated in the transition zone and the central zone of the hyperplastic prostate. Stage A2 cancers were mostly middle grade to high grade cancers initiated in the peripheral region and then invaded the central area. The lower incidence of stage A carcinoma in China is related to the small amount of biopsy tissue, careless microscopic observation, or lack of the ability to diagnose stage A1 cancer. PMID- 11866983 TI - [A clinicopathological study of clear cell sarcoma of the kidney]. AB - OBJECTIVE: To study the clinicopathological, immunohistochemical features and the histogenesis of clear cell sarcoma of the kidney (CCSK). METHODS: CCSK specimens from 45 pediatric cases, including 31 male and 14 female with an age range from 3 months to 12 years (mean of 3.2 years), were retrieved. Routine pathological, immunohistochemical and electron microscopic methods were utilized to analyze the CCSK specimens. RESULTS: 35 of the 45 cases were followed from 6 to 192 months. 15 patients presented with bone metastases, 6 had lung or liver metastases, 8 recurred and 20 died. Age and clinical stage at diagnosis correlated with the rate of survival. Histologically, the classic pattern of CCSK consisted of cells with pale cytoplasm, fine nuclear chromatin and indistinct nucleoli separated by an arborizing fibrovascular stroma. Other patterns were identified, including myxoid, spindle, palisading, epithelioid, sclerosing, cellular, cystic, and angiectatic. All tumors contained multiple patterns. Immunohistochemically, all cases were positive for vimentin, but negative for EMA, CK, desmin, actin, S-100, NSE, CD99, CD34 and LCA. Electron microscopy of 9 cases showed features of primitive cell conjunction and few organelles. CONCLUSION: CCSK is a common renal neoplasm of childhood. CCSK may arise from renal mesenchymal cells and has the propensity to metastasize to the bone with poor clinical outcome. PMID- 11866984 TI - [Detection of SYT-SSX fusion gene in paraffin-embedded tissues and its clinicopathologic significance for synovial sarcoma]. AB - OBJECTIVE: To investigate the feasibility of detecting SYT-SSX fusion gene in paraffin-embedded tissues and its diagnostic significance for synovial sarcoma (SS). METHODS: Formalin-fixed, paraffin-embedded samples of 38 cases of SS and 40 cases of control tumors (including spindle cell sarcoma and metastatic adenocarcinoma) were retrived from archival materials. SYT-SSX fusion transcripts were detected in all samples by RT-PCR. House-keeping gene Porphobilinogen Deaminase (PBGD) was used to detect the quality of mRNA. RESULTS: PBGD mRNA was detected in 64 of the 78 tumor cases (82.1%). SYT-SSX fusion transcripts was detected in 33 of the 38 synovial sarcoma specimens. No SYT-SSX mRNA expression was found in control tumors. SYT-SSX mRNA expression rate reached 89.2% (33/37) in synovial sarcomas after exclusion of 1 case which was negative for both SYT SSX and PBGD. In 33 SYT-SSX positive synovial sarcomas, 22 had SYT-SSX1 and 6 had SYT-SSX2 fusion transcripts. In 5 cases, the fusion type could not be distinguished. There was an association between SYT-SSX fusion type and histologic subtype. All 10 biphasic synovial sarcoma cases had the SYT-SSX1 fusion, whereas 12 of 18 monophasic synovial sarcoma had the SYT-SSX1 and 6 had the SYT-SSX2 fusion gene (P < 0.05). CONCLUSIONS: (1) Detection of SYT-SSX fusion gene in paraffin-embedded tissues was found to be a sensitive and specific method for the diagnosis and differential diagnosis of synovial sarcoma. (2) There was an association between SYT-SSX fusion type and histologic subtype.SYT-SSX2 fusion transcript could only be found in monophasic synovial sarcomas. PMID- 11866985 TI - [Detection and analysis of SYT-SSX fusion gene in synovial sarcoma]. AB - OBJECTIVE: To demonstrate that a characteristic SYT-SSX fusion gene resulting from chromosomal translocation t (X; 18) (p11; q11) was detectable in archival paraffin-embedded synovial sarcoma tissues. METHODS: SYT-SSX fusion transcripts in 20 synovial sarcomas (histologic subtypes, 15 monophasic and 5 biphasic) were detected and analyzed by the reverse-transcriptase polymerase chain reaction and compared with the results from relevant pathological data. RESULTS: A specific SYT-SSX RT-PCR product was found in 19 of 20 (95%) synovial sarcomas tested and of the 13 tumors containing SYT-SSX2, 10 were monophasic. CONCLUSION: SYT-SSX fusion transcripts are considered as a defining diagnostic marker of synovial sarcomas and the subtypes of SYT-SSX fusion transcripts (SYT-SSX1 and SYT-SSX2) may yield prognostic information. PMID- 11866986 TI - [Study on expression of caspase-3 and bcl-2 proteins and their relations with cell apoptosis and proliferation in non-Hodgkin's lymphoma]. AB - OBJECTIVE: To explore the possible roles of caspase-3 and bcl-2 proteins in the development and progression of non-Hodgkin's lymphoma (NHL) and their relationship. METHODS: TdT-mediated dUTP nick end labeling (TUNEL) and immunohistochemistry were used to study cell apoptosis and expression of proliferating cell nuclear antigen (PCNA), caspase-3 and bcl-2 in 119 NHL cases. RESULTS: The caspase-3 and bcl-2 positive rates were 86.6% (103/119) and 53.8% (64/119) respectively. Contrasting expression of these two proteins were found in reactive and malignant lymphoid tissues: In reactive lymphoid follicles, caspases 3 exhibited strong immunoactivity in cells located in germinal centers but not in mantle zone lymphocytes, whereas bcl-2 exhibited strong immunoactivity in mantle zone lymphocytes but not in cells located in germinal centers, and in neoplastic follicles, caspase-3 was negatively or weakly expressed, whereas bcl-2 was often positively expressed. In B cell lymphomas, the high grade group had a higher caspase-3 immunointensity and a lower bcl-2 immunointensity than the low grade group (P < 0.01). Apoptotic index correlated positively with expression of caspase-3 (r = 0.512, P < 0.01) and inversely with expression of bcl-2 (r = 0.436, P < 0.01). In addition, apoptotic index and proliferative index were positively related (r = 0.710, P < 0.01). CONCLUSIONS: caspase-3 may participate in the regulation mechanism of lymphoma cell apoptosis. The contrasting expression of caspase-3 and bcl-2 is often seen in reactive and malignant lymphoid tissues, and it may indicate a close relationship between these two proteins in the regulation of lymphocyte proliferation kinetics. PMID- 11866987 TI - [Detection of the expression of Smad4, transforming growth factor beta(1) and beta receptor II proteins in paraffin-embedded human pancreatic cancer tissues]. AB - OBJECTIVE: To explore the relationships between Smad4, TGF beta1 and TbetaR II in the TGF-beta pathway and the possible mechanisms by which they effect pancreatic carcinoma. METHODS: The expression of Smad4, TGF beta1 and TbetaR II in paraffin embedded pancreatic carcinoma tissues was detected by using antibodies against Smad4, TGF beta1 and TbetaR II with EnVision immunohistochemistry. RESULTS: The positive rates of Smad4, TGF beta1 and TbetaR II were 58.93% (33), 66.07% (37) and 60.71% (34), respectively. The positive rates of the above three proteins in matched normal pancreatic tissues were 89.29% (50), 25.00% (14) and 25.00% (14) respectively. There was a significant relationship between the expression of TGF beta1, clinical stage and the metastasis of the tumor (P < 0.05). There was also a significant relationship between the expressions of TGF beta1 and TbetaR II (P < 0.05). CONCLUSIONS: The expression of Smad4 in pancreatic carcinoma tissue is significantly decreased but the expression of TGF beta1 and TbetaR II are increased. Smad4 may play an important role in the regulation of TGFbeta inducible gene expression and subsequent growth inhibition, but TGFbeta may also act in a Smad4-independent manner. PMID- 11866988 TI - [Effect of hepatitis C virus nonstructural protein NS3 on telomerase activity]. AB - OBJECTIVE: To study the effect of hepatitis C virus nonstructural protein NS3 (HCV NS3) on telomerase activity and its carcinogenesis, and to observe the practical use of in situ telomerase activity labeling. METHODS: NIH3T3 cells were transfected with plasmid pRcHCNS3-5prime prime or minute (expressing HCV NS3 C terminal deleted protein), pRcHCNS3-3prime prime or minute (expressing HCV NS3 N terminal deleted protein) and blank plasmid pRcCMV. Positive clones, 11 with plasmid pRcHCNS3-5prime prime or minute, 11 with pRcHCNS3-3prime prime or minute and 8 with plasmid pRcCMV were harvested respectively. Streptavidin-peroxidase conjugated method (SP) was used to detect the expression of HCV NS3 protein in NIH3T3 cells transfected with plasmid pRcHCNS3-5prime prime or minute and pRcHCNS3-3prime prime or minute. Telomerase activity was detected by in situ telomerase activity labeling method and telomerase PCR ELISA technique in the transfected and non-transfected NIH3T3 cells. RESULTS: HCV NS3 protein was expressed in the cells transfected with plasmid pRcHCNS3-5prime prime or minute or plasmid pRcHCNS3-3prime prime or minute. The positive signals of HCV NS3 protein were localized in the cytoplasm of transfected NIH3T3 cells, and the signal intensity of the former was stronger (chi(2) = 6.667 P < 0.05). There was significant difference on telomerase activity between each group (F = 134.083 P < 0.01). Telomerase activity in all 11 clones with plasmid pRcHCNS3-5prime prime or minute was stronger than cells with plasmid pRcHCNS3-3prime prime or minute (P < 0.01), whereas telomerase activity in NIH3T3 cells transfected with the plasmid pRcCMV or non-transfected NIH3T3 cells were weaker than the cells with the plasmid pRcHCNS3-3prime prime or minute. The expression level of HCV NS3 protein were correlated significantly with the strength of telomerase activity (rs = 0.8084 P < 0.01). The results obtained with in situ telomerase activity labeling corresponded to those with telomerase PCR ELISA technique (rs = 0.50196 P < 0.01). CONCLUSIONS: (1) HCV NS3 protein may activate telomerase through endogenous mechanism to induce host cells transformation. (2) The effect of HCV NS3 C-terminal deleted protein on telomerase activity in host cells may be more intense than that of HCV NS3 N-terminal deleted protein. (3) In situ telomerase activity labeling is a reliable technique for studying pathological morphology and telomerase activity in tissues and cells. PMID- 11866989 TI - [Studies of intercellular communication in human rhabdomyosarcoma cell lines of different metastatic potential]. AB - OBJECTIVE: To investigate the relationship between intercellular communication and three human rhabdomyosarcoma (RMS) cell lines of different metastatic potential. METHODS: Indirect immunofluorescent staining and laser scanning confocal microscope (ISCM) were used to detect connexin 43 (CX43), a molecule related with gap junctional intercellular communication (GJIC). Fluorescence redistribution after photobleaching (FRAP) was used to detect function of GJIC. RESULTS: A high level of CX43 protein expression was revealed in normal human myoblasts. CX43 protein was mainly localized on the surface of cells and sometimes in cytoplasm. CX43 expression was decreased in RMS cells. Both the detection rates and fluorescent intensity of CX43 decreased when the metastatic potential of RMS increased (P < 0.05). In contrast to normal, the fluorescence recovery rates of the RMS cells decreased and there was a negative correlation between the function of GJIC and malignancy of RMS cell lines (P < 0.05). CONCLUSION: Varying degrees of GJIC inhibition may correlate with different metastatic potential of RMS. This may help to determine the malignant behavior of RMS and be used as a prognostic index of RMS. PMID- 11866992 TI - Editorial: Herpesvirus Infections in Transplant Recipients. PMID- 11866990 TI - [Expression of matrix metalloproteinase-2 in rat pulmonary fibrosis]. AB - OBJECTIVE: To study the role of matrix metalloproteinase-2 (MMP(2)) in rat pulmonary fibrosis. METHODS: (1) Dynamic changes of MMP(2) expression were observed in bleomycin-induced rat pulmonary fibrosis by use of immunohistochemical technique. (2) RT-PCR was carried out to show the dynamic changes of MMP(2) mRNA expression of isolated pulmonary fibroblasts (PFbs) during rat pulmonary fibrosis. (3) Northern blot analysis and immunocyto chemistry techniques were performed to detect the expression of MMP(2) mRNA and their relevant protein in cultured rat PFbs after treatment with transforming growth factor 1 (TGF beta(1)). RESULTS: (1) On day 1-7th after receiving bleomycin, the infiltrated pulmonary macrophages and septa mesenchymal cells showed positive reaction with anti-MMP(2) and an increase in number. After 14 days and on 28 th of bleomycin treatment, the number of positive staining macrophages and mesenchymal cells were decreased respectively. (2) The gene expression of MMP(2) mRNA of PFbs were enhanced after bleomycin treatment from the Day 1 and decreased by Day 28. (3) The expression of MMP(2) mRNA and their relevant protein were enhanced in rat PFbs after TGF beta(1) treatment. CONCLUSIONS: Over expression of MMP(2) in pulmonary tissue may be an important factor in the initiation of pulmonary fibrogenesis. PMID- 11866991 TI - [Study on the correlation between the expression of protein p53 at stage III nasopharyngeal carcinoma and its biological behavior, prognosis, p53 gene mutation and the expression of LMP-1]. AB - OBJECTIVE: To investigate p53 protein expression at stage III nasopharyngeal differentiated non-keratinizing squamous carcinoma (DNKSC) and their correlation with biological behavior and prognosis;the relationship between p53 protein expression and p53 gene mutation and LMP-1 protein expression at 58 cases of stage III DNKSC. METHODS: Immunohistochemistry (Envision) was applied to detect p53 protein and LMP-1 protein expression. Non-isotopic PCR-SSCP was used to detect p53 gene mutation. RESULTS: The p53 protein expression showed general positive rates of 65.5% (38/58) in the patients with nasopharyngeal DNKSC, being significantly higher in the group surviving less than 5 years 82.1% (32/39) than in the group surviving longer than 5 years 31.6% (6/19) (P < 0.05). Besides, it was closely correlated with the local invasion to the cranial base, there was a significant difference between invasion 74.4% (29/39) and non-invasion 47.4% (9/19) (P < 0.05). The expression of LMP-1 protein rated 72.4% (42/58), exhibiting a positive correlation with the expression of p53 protein (r = 0.504). The p53 gene mutation was found in none of the cases (0/15). CONCLUSIONS: In stage III nasopharyngeal DNKSC, the expression of p53 protein is not related to p53 gene mutation, whereas it relates to the expression of LMP-1 protein. Declined expression of p53 protein is closely related to the time of survival, and the p53 protein expression is higher in the invasive carcinoma. PMID- 11866993 TI - Post-transplantation Lymphoproliferative Disease. AB - Post-transplant lymphoproliferative disorders are a varied group of disease processes that follow both solid organ and bone marrow transplantation. The disorders are typically associated with the Epstein-Barr virus and, as a consequence, have contributed greatly to our understanding of the role of human viruses in the pathogenesis of malignant disease. They have also expanded our understanding of the role of immunosurveillance in the prevention and control of malignancies which has, in turn, led to exciting developments in the use of transferred or adoptive immunotherapy for the treatment of cancer. This technology holds great potential for the treatment of these and other diseases associated with immunodeficiency states. When recognized early in their natural history, the post-transplant lymphoproliferative disorders appear to be curable; later detection frequently leads to a fatal outcome. Prompt diagnosis of these disorders by the clinician caring for the transplant patient is therefore critical. PMID- 11866994 TI - Adoptive Transfer of Herpesvirusspecific Cytotoxic T Lymphocytes in Transplant Recipients. AB - Profound suppression of T-cell immunity observed after transplantation may permit the emergence of life-threatening viral complications. Prevention or treatment of viral infections in immunocompromised patients through the infusion of small numbers of antigen-specific T-cell lines or clones is the most successful example of adoptive immunotherapy in humans. In fact, adoptive transfer of T cells of donor origin has been demonstrated to be able to restore protective immunity towards these infectious agents and to control established disease. This article reviews human trials of adoptive immunotherapy for herpesvirus-related complications, with the aim of defining principles and problems encountered so far to aid the design of future clinical studies. PMID- 11866995 TI - CMV Infection in Bone Marrow and Solid Organ Transplant Patients in the Era of Antiviral Prophylaxis. AB - Recent developments in the diagnosis and therapy of cytomegalovirus (CMV) infection have helped to reduce CMV-associated morbidity and mortality following allogeneic bone marrow and solid organ transplantation. The clinical symptoms of active CMV infection and the prevalence of life-threatening CMV disease vary widely between different patient populations according to the type of transplant and the intensity of immunosuppression employed. Antiviral prophylaxis with aciclovir, valaciclovir and ganciclovir has been shown to reduce CMV infection and disease following organ transplantation. Antiviral drugs, in particular ganciclovir and foscarnet, have varying sideeffects, however, and antiviral resistance due to prolonged administration of ganciclovir and foscarnet has been reported recently. Short courses of pre-emptive antiviral therapy for documented CMV infection help to reduce the duration and sideeffects of therapy, offering an alternative strategy to antiviral prophylaxis. Studies are required to compare the efficacy and costs of antiviral prophylaxis with pre-emptive therapy. PMID- 11866996 TI - Kaposi's Sarcoma and Transplantation. AB - Kaposi's sarcoma is a tumour of multicentric origin, composed of endothelium lined vascular spaces and spindle-shaped cells. The incidence of Kaposi's sarcoma in transplant recipients is 400--500 times greater than that in the general population, and is rising within the transplant population, currently comprising more than 5% of all de novo neoplasms in this group. The exact pathogenesis is still unknown but DNA sequences from human herpesvirus 8 (HHV-8) are present in the different clinical variants of Kaposi's sarcoma. Risk factors associated with development of these tumours posttransplantation include the geographical origin of the patient, HSV-8 infection before and after transplantation, and the immunosuppressive regimen used, but the importance of each factor remains to be determined. Apart from conventional treatment with surgical excision, radiotherapy or chemotherapy, cessation, reduction or modification of immunosuppressive therapy has been shown to be effective in a significant number of patients. This article reviews recent advances in our understanding of Kaposi's sarcoma after transplantation. PMID- 11866997 TI - Post-transplant HHV-6 Diseases. AB - Human herpesvirus 6 (HHV-6) infection is common in the general population. Primary infection occurs mainly in childhood, after which the virus establishes latency in the host and can be reactivated in immunocompromised individuals such as recipients of a solid organ or bone marrow transplant (BMT). HHV-6 infection occurs in 38--60% of BMT recipients and in 14--82% of solid organ recipients, usually 2--4 weeks post-transplantation. Two HHV-6 variants are recognized (HHV 6A and HHV-6B) and transplant patients are infected almost exclusively with variant B. Following BMT, the clinical diseases most commonly associated with HHV 6 infection are bone marrow suppression, interstitial pneumonitis, encephalitis and graft versus host disease. It has also been suggested that HHV-6 infection may be associated with fever and bone marrow suppression following liver transplantation and may contribute to rejection episodes after kidney transplantation. Infection with HHV-6 induces marked immunodepression, and an association both with cytomegalovirus disease and with fungal infection in transplant recipients has been reported. A direct causal relationship between HHV 6 infection and disease following transplantation, however, has yet to be proven conclusively. PMID- 11866998 TI - Editorial: Controversies in Herpesvirus Infections: Implications for Clinical Management. PMID- 11866999 TI - Human Herpesvirus 6 and Febrile Convulsions. AB - Human herpesvirus 6 (HHV-6) is a ubiquitous virus that causes the childhood febrile illness exanthem subitum. Primary infection usually occurs in the first few years of life and the virus is considered to be one of the most common causes of acute febrile illness in childhood. HHV-6 can infect the central nervous system, causing meningitis or encephalitis after an acute attack of exanthem subitum. Febrile convulsions are common in early childhood, affecting 2--3% of children between 6 months and 5 years of age. Primary HHV-6 infection has been reported to be complicated by febrile convulsions in 8--50% of cases. An association between HHV-6 and febrile convulsions was established on the basis of: the similarity between the age groups in which febrile convulsions and HHV-6 infection occur, the high incidence of febrile convulsions after HHV-6 infection and the neurotropic properties of the virus. The exact role of HHV-6 in the pathogenesis of febrile convulsions remains unclear. PMID- 11867000 TI - Human Herpesvirus 8 and Protection from AIDS Dementia Complex. AB - Infection with human herpesvirus 8 (HHV-8) has been associated with the development of three distinct conditions: Kaposi's sarcoma, body cavitybased lymphoma and Castleman's disease. HHV-8 produces chemokinelike proteins including viral macrophage inflammatory protein II, which has been shown to block human immunodeficiency virus 1 (HIV-1) infection of CD4-positive cells expressing CCR 3. As CCR-3 is a receptor for HIV-1 into microglial cells, it has been hypothesized that HHV-8 infection may inhibit HIV-1 infection of the brain, thereby decreasing the incidence of AIDS dementia complex. We reviewed published studies of the incidence of AIDS dementia complex in individuals with and without Kaposi's sarcoma. The data are consistent in showing a negative association between Kaposi's sarcoma and AIDS dementia complex and, although sparse, support the hypothesis that productive HHV-8 infection decreases HIV-1 infection of the brain sufficiently to decrease the incidence of AIDS dementia complex. This negative association should be examined in further cohorts of HIV-1-infected subjects, to exclude alternative explanations. PMID- 11867001 TI - Chronic Fatigue Syndrome and Herpesviruses: the Fading Evidence. AB - Herpesviruses, in particular Epstein-Barr virus (EBV), cytomegalovirus (CMV) and human herpesvirus 6 (HHV-6), have, for the past two decades, come under considerable scrutiny as aetiological agents of chronic fatigue syndrome (CFS). However, virological findings of herpesviruses in CFS have not been consistent between different studies, and the unusual patterns of serological responses to EBV, CMV and HHV-6 have not been specific for CFS, being observed also in asymptomatic individuals. In addition, patients with symptomatology suggestive of CFS do not appear to have an increased frequency of these herpesviruses, as detected by culture or polymerase chain reaction, compared with controls, which argues against an ongoing active herpetic infection. Studies have also shown that the presumable elevation of antibody titres to EBV, CMV or HHV-6 in CFS are not observed only with these viruses, but also with other organisms such as herpes simplex virus and measles. PMID- 11867002 TI - Does Cytomegalovirus Play a Role in Atherosclerosis? AB - In vitro experiments, as well as experiments using animal models, suggest that cytomegalovirus (CMV) infection can stimulate atherosclerosis. There is also evidence that CMV stimulates the development of restenosis and transplantation associated arteriosclerosis. Activation of arterial cells, enhancement of the inflammatory response and induction of cytokine production may all be important in mediating these effects. Further experiments, especially in animals susceptible to atherosclerosis and in patients with cardiovascular disease, will be necessary to elucidate the role of CMV, and also that of other infectious agents, in atherosclerosis. PMID- 11867003 TI - Editorial: Our Evolving Knowledge of Herpesviruses. PMID- 11867004 TI - History of Varicella Zoster Virus. AB - The earliest reports of vesicular rashes of the type we now recognize to be caused by herpes simplex and zoster date to the ancient civilizations. It was not until 1888, however, that a relationship between herpes zoster and chickenpox was suggested. Establishing this link represented one of the major hurdles in the history of varicella zoster virus. There was no animal host and this meant that much of the evidence needed to be obtained by clinical and epidemiological observation. Since the link was proven, in the 1950s, the advent of the live attenuated vaccine virus, in 1974, and aciclovir in the 1980s, has had a huge impact on prevention and treatment, respectively. The complete DNA sequence of VZV was established in 1986. A more complete understanding of the VZV genome and its gene products may enable recombinant vaccines and specific therapies to be advanced. We also need to determine the long-term effects of the use of the varicella vaccine. The ultimate aim: to prevent VZV infection completely. PMID- 11867005 TI - Cytomegaloviruses: a Historical Perspective. AB - The cellular changes associated with the cytomegaloviruses result in enlarged cells with prominent nuclei. These were first described by pathologists and for many years they were considered to be parasitic protozoa. In 1926, however, it was suggested that the aetiological agent was probably a virus, and the viruses responsible were isolated independently by three groups in 1956. Demonstration of antigenic heterogeneity between the isolates led to the term cytomegaloviruses (CMV). Clinical studies first concentrated on overt congenital infection in CMV infected individuals who exhibited neurological defects; however, the picture of congenital infection was altered and expanded by subsequent longitudinal studies. Asymptomatic CMV-infected newborns were often found to develop deafness. It was discovered that infected children could excrete CMV in their urine for several months and, from this source, the virus was able to spread rapidly in environments such as day-care centres. While CMV infection acquired by adults was often asymptomatic, a CMV mononucleosis syndrome was later recognized. This was followed, in 1966, by the discovery that CMV mononucleosis could develop after the transfusion of CMV-infected blood. Subsequently, CMV infection in the immunosuppressed subject became recognized as a serious cause of morbidity and mortality. PMID- 11867006 TI - Human Herpesvirus 6: an Evolving Story. AB - First isolated in 1986 from patients with lymphoproliferative disorders, human herpesvirus 6 (HHV-6) is a B-herpesvirus with two variants: HHV-6A and HHV-6B. HHV-6B is the major causal pathogen of exanthem subitum, a predominantly benign exanthematous disease of infants with occasional complications in the central nervous system. Infection with HHV-6 is common among the general population and the virus mainly seems to be transmitted from mother to infant via saliva. Following primary infection, HHV-6 persists latently and can reactivate in immunocompromised hosts, such as in individuals with AIDS or in transplant or multiple sclerosis patients where it possibly causes encephalitis and pneumonitis. Its precise role in these conditions is not well understood and further study is needed. PMID- 11867007 TI - An Overview of Epstein-Barr Virus: from Discovery to Future Directions for Treatment and Prevention. AB - Discovered in 1964 by Epstein and Barr, the Epstein-Barr virus (EBV) is widespread in all areas of the world, infecting over 95% of the adult population and earning it the informal name, 'Every Body's Virus'. It was shown to be the causal agent of infectious mononucleosis (IM) in 1968, and many reports followed linking EBV to several other clinical syndromes. In 1970, the virus was shown to be able to immortalize B lymphocytes, which are one of its target cells. This oncogenic potential underlies the role of EBV in Burkitt's lymphoma, post transplant lymphoproliferative disorders and lymphoma in HIV-infected patients. In addition, EBV can infect epithelial cells in the nasopharynx and is a co factor for development of nasopharyngeal carcinoma. There is no treatment available for IM to date; however, virus-associated complications may be treated with various modes of immune modulation. Currently, radiotherapy and chemotherapy remain the main treatments for EBV-associated malignancies. Recent evidence has suggested that adoptive therapy through infusions of human leukocyte-associated antigen-matched EBV cytotoxic T cells may form a novel strategy for both prophylaxis and treatment of EBV-induced lymphoproliferative disorders. Additionally, a vaccine based on immunization with a structural antigen, gp350, is under evaluation. PMID- 11867009 TI - Use of type-specific HSV serology in the management of a 37-year-old woman diagnosed with primary genital herpes. AB - A 37-year-old woman attended a Genitourinary Medicine Clinic in London, UK, concerned about the reliability of a culture-based diagnosis of genital herpes that she had received while abroad. She was in a longterm monogamous relationship with a partner who recalled no history of genital herpes. She was unsure as to how she had acquired genital herpes and wished to know whether she was newly infected. Typespecific serological testing indicated a true primary herpes simplextype-2 (HSV-2) infection. The patient's partner did not attend the clinic but reported a (negative blood test for herpes) at a clinic elsewhere. No details were provided as to the type of assay used, so the possibility that this result was inaccurate cannot be ruled out. It is possible that he may have been an asymptomatic carrier of HSV (either HSV-2 or both HSV-1 and-2) or had mild unrecognized symptoms of genital HSV. This case highlights the advantages and pitfalls of using a type-specific serological test for the diagnosis of genital herpes and also the need for appropriate counselling. An initial explanation of the significance of the HSV culture result she received abroad may have reduced the patient's anxiety about the reliability of the initial diagnosis. PMID- 11867010 TI - Potential for immunotherapy in the treatment of herpesvirus infections. AB - The concept of using immunotherapy to treat recurrent herpesvirus infections dates back to the 1930s, although many of the initial studies were seriously flawed. Since the late-1980s, however, the use of the guinea pig model of genital herpes has allowed investigators to evaluate carefully several vaccine and immunomodulatory strategies for the control of recurrent herpesvirus infections. These investigations have clearly shown that both approaches can significantly decrease recurrence rates of genital herpes, and the most effective routes, adjuvants and treatment regimens have been identified. Similar strategies have also been shown to decrease herpes simplex virus (HSV)-1 recurrences in animal models of ocular infection. To date, only moderate success has been reported for human trials, although the optimum strategies that were identified in the animal models have not yet been evaluated. PMID- 11867011 TI - Recent developments in herpesvirus therapy. AB - The antiherpes drugs, aciclovir and ganciclovir, are considered the standard treatments and prophylactic agents for infections caused by herpes simplex virus (HSV), varicella zoster virus (VZV) and cytomegalovirus (CMV). Until a decade ago, the impact of aciclovir on the control of severe and life-threatening herpesvirus infections was unprecedented. During the past few years, we have witnessed approval of new therapeutic drugs for infections caused by HSV and VZV (i.e. penciclovir and the oral prodrugs, valaciclovir and famciclovir), CMV (i.e. ganciclovir, cidofovir and fomivirsen) or HSV, VZV and CMV (i.e. foscarnet). A few agents, such as brivudin and benzimidavir, are in ongoing clinical development; others have been suspended because of safety concerns. New antiherpes agents are needed to face clinical issues such as drug resistance, increased use of antiherpes prophylaxis in transplantation and safety concerns in small children or pregnant women. PMID- 11867012 TI - Genetically engineered human herpes simplex virus in the treatment of brain tumours. AB - Central nervous system malignancies--particularly glioblastoma multiforme--pose significant problems for the development of novel therapeutics. In the absence of advances with standard surgical and chemotherapeutic approaches, the utilization of genetically engineered viruses--both as direct oncolytic agents (virus therapy) and for the delivery of foreign proteins (gene therapy)--represents a significant advance in the experimental approach to the management of patients with incurable tumours. Among other viruses, herpes simplex virus (HSV) offers an opportunity to influence the replication of tumour cells directly within the central nervous system. The propensity for HSV to replicate in tumour cells, and its large coding capacity, provide an experimental model for the development of novel therapeutics. The status of these experimental approaches and Phase I studies are summarized. PMID- 11867013 TI - The nine ages of herpes simplex virus. AB - Hippocrates was the first to describe lesions that could have been caused by herpes simplex virus (HSV), but the clinical conditions caused by this virus have only been described in more detail over the past 3 centuries. Most of the key findings relating to HSV infection and treatment have been made since the early 20th Century. These range from discovering some of the mechanisms behind virus latency and reactivation, to the development of the drug aciclovir, which was the first selective inhibitor of HSV replication. This review provides an evolutionary understanding of HSV, but HSV research is still in its golden age. New facts continue to emerge about HSV, and manipulation of the virus is providing much information. Genetic engineering of this virus is likely to have a most significant impact on future medical therapies, which could extend to specialties beyond virology. PMID- 11867016 TI - Changing trends in cytomegalovirus disease in HIV-infected patients. AB - Treatment with combination antiretroviral therapy has transformed the natural history of human immunodeficiency virus (HIV) infection. Effective antiretroviral therapy results in both reduced risk of cytomegalovirus (CMV) disease and prolonged survival following CMV disease. These effects seem to be mediated by the reconstitution of immune responses against CMV, which results in re established host control of CMV replication. As a result, some individuals have been able to discontinue maintenance therapy for CMV with a low risk of disease recurrence to date. The risk of development or progression reappears in the context of antiretroviral failure--a problem increasingly recognized in clinical practice. However, improved immune function is not always beneficial. Indeed, previously uncommon inflammatory complications have been associated with immune reconstitution following highly active antiretroviral therapy. The natural history of CMV disease at a time of rapid changes in treatment strategies remains uncertain and, therefore, individuals at risk continue to require careful management. PMID- 11867015 TI - Prevention and treatment of VZV infections in patients with HIV. AB - Varicella zoster virus (VZV) infections in human immunodeficiency virus (HIV) infected patients are known to have a different disease spectrum from that seen in other types of patients. Varicella in children with HIV infection is likely to be more serious than in otherwise healthy children and routine antiviral therapy is recommended. There is evidence that the development of varicella in HIV infected children is not associated with progression to AIDS, suggesting that it may be safe to immunize HIV-infected children with live attenuated varicella vaccine. There are no published data on varicella in HIV-infected adults, however, probably because most adults have already experienced varicella prior to HIV infection. Zoster in HIV-infected children differs somewhat from that in HIV infected adults. In particular, HIV-infected children who develop varicella in the setting of severe immunodeficiency are at an especially high risk to develop zoster. Given the low rate of toxicity of aciclovir as well as its ease of administration and its efficacy in hastening the healing of VZV infections, prompt treatment with this antiviral agent is recommended for both HIV-infected children and adults. Foscarnet should be used for zoster that is strongly suspected or proven to be caused by aciclovir-resistant VZV. Patients with HIV for whom there is no evidence of significant immunosuppression and who have not had varicella should be immunized with live attenuated varicella vaccine as a preventative measure for both varicella and zoster. It is hoped that immunization of VZV seropositive HIV-infected patients will decrease the incidence of zoster. Studies to determine this are under way. PMID- 11867017 TI - Management of genital herpes in HIV-infected patients. AB - The management of genital herpes in patients infected with human immunodeficiency virus (HIV) differs from that in individuals with genital herpes because of the significant interaction between the two viruses involved. HIV-induced immunodeficiency increases the frequency and severity of recurrent anogenital herpes simplex virus (HSV) shedding and disease as well as the risk of developing drug-resistant HSV infection. HSV infection, in turn, increases HIV replication and the risk of HIV transmission. The advent of highly active antiretroviral therapy has facilitated therapy for genital herpes, but important unanswered questions remain about the optimal therapy of drug-sensitive and -resistant genital herpes and the role of antiherpes drugs in reducing HIV disease progression and the risk of HIV transmission. PMID- 11867018 TI - The role of HSV in the transmission and progression of HIV. AB - Herpes simplex virus (HSV) is a common co-infection in persons infected with human immunodeficiency virus type 1 (HIV-1). Chronic perianal ulceration from herpes simplex virus type 2 (HSV-2) was one of the first opportunisitc infections identified among patients with AIDS. Subsequent research has established that the natural history of HSV-2 is altered in co-infected persons as they experience more frequent clinical and subclinical reactivation of HSV than persons infected only with HSV-2. In addition, there are accumulating data to suggest a significant biological interaction between these two viruses that result in more efficient sexual transmission of HIV-1 and an increased rate of HIV replication during both clinical and subclinical HSV reactivation. PMID- 11867019 TI - Molecular basis of the interactions between herpes simplex viruses and HIV-1. AB - Herpes simplex virus types 1 and 2 (HSV-1 and -2) are two of the major opportunistic agents involved in the pathogenesis of AIDS, which is caused by human immunodeficiency virus types 1 and 2 (HIV-1 and HIV-2). A body of evidence suggests that they can also act as co-factors by interacting with HIV-1, thereby influencing disease progression. Indeed, the HIV-1 life cycle can be affected by HSV at different levels of interaction, both in vitro and in vivo: (i) transactivation of the HIV-1 long terminal repeat can be mediated, probably through different pathways, by HSV-1-infected cell protein (ICP)0, ICP4, ICP27 and US11 gene products; the HSV-1 transactivator viral protein 16 is not able to transactivate the long terminal repeat; (ii) cytokine release and antigen presentation from HSV-infected cells are both able to stimulate HIV-1 expression; (iii) Pseudotyping of the HIV-1 core particle with HSV-1 envelope glycoproteins can expand HIV-1 tropism to new cell types. Moreover, in vivo studies report that aciclovir treatment can produce a survival benefit in HIV-1-infected patients and that recurrent genital herpes appears to be linked to HIV-1 transmission by both boosting plasma retroviral load and providing a portal of entry and exit for HIV 1. PMID- 11867021 TI - Herpesvirus and multiple sclerosis. AB - Evidence suggests that multiple sclerosis (MS) is caused by host genetic factors in association with one or more environmental agents. The clinical and pathological features of MS implicate viral infections as either cofactors in its aetiology or triggers of relapses, although no specific environmental factors have been identified. In particular, several herpesviruses have attracted interest because their ability to cause latent infections that periodically reactivate has some familiarity with the relapsing-remitting course of MS. Further, most human herpesviruses can be readily found within the central nervous system and several are known to be capable of inducing demyelination, both in humans and in experimentally infected animals. This brief review takes a systematic look at reported associations between herpesviruses and MS and suggests some criteria that must be met in further studies. One of the greatest challenges in confirming or refuting a role for herpesviruses in chronic diseases is their ubiquitous nature. It is stated from the outset that the associations described here remain controversial. PMID- 11867022 TI - The association of herpes simplex virus and Alzheimer's disease: a potential synthesis of genetic and environmental factors. AB - The chronic and sporadic nature of Alzheimer's disease (AD), in which the majority of presenting cases are over 65 years of age, suggests that compounded injury or long-term exposure to environmental factors is involved in the aetiology, rather than inherited genetic alterations. Great hope has been placed on identifying a causative infectious agent that would present the potential for therapeutic or preventative vaccines or drugs. Although not the sole causative agent, herpes simplex virus type 1 has been the most clearly established pathogen to be associated with AD development to date. Recent advances have begun to suggest mechanisms by which this ubiquitous, neurotropic pathogen may predispose an individual to the development of AD. Such progress should ultimately lead to therapeutic strategies that limit predisposing neurological damage and potentially reverse the process, increasing the quality and quantity of life in this patient population. PMID- 11867023 TI - Acute retinal necrosis: insights into pathogenesis from the mouse model. AB - Acute retinal necrosis (ARN) is a relatively rare syndrome that is caused by infection with one of several members of the human herpesvirus family. ARN usually occurs in otherwise healthy patients, although it has also been observed in immunocompromised individuals. It is characterized by retinal vasculitis and haemorrhaging, areas of retinal necrosis, vitreous and aqueous inflammation and optic neuritis. It may affect one or both eyes and frequently results in severely reduced vision or blindness in the affected eye. Results using the mouse model of ARN have provided insight into the pathogenesis of this disease. However, many unanswered questions remain, such as why does only a very small fraction of individuals infected with one or more herpesvirus develop ARN? Increased understanding of the interactions of herpesviruses with T cells and cytokines may enable the development of therapeutic strategies targeted specifically to control viral infection in the eye and/or brain. PMID- 11867024 TI - Serological testing for human herpesvirus 8. AB - Human herpesvirus 8 (HHV-8) has been identified in skin lesions of patients with Kaposi's sarcoma and is now considered a co-factor for the development of the neoplasm associated with this disease. Several laboratory tests are currently available for use in the detection of HHV-8 antibodies--immunofluorescence and enzyme-linked immunosorbant assays being the most frequently used tests. Epidemiological studies have demonstrated the existence of hyperendemic regions where HHV-8 infection is ubiquitous and often acquired early in childhood. The role of HHV-8 in the development of Kaposi's sarcoma has yet to be clarified completely and data collected from the epidemiological studies of HHV-8 prevalence may be useful in gaining a better understanding of the different factors involved in tumour development. PMID- 11867025 TI - Case study: rapid emergence of a cytomegalovirus UL97 mutant in a heart transplant recipient on pre-emptive ganciclovir therapy. AB - A cytomegalovirus (CMV) strain containing a mutation in the UL97 gene (Leu595Ser), which is known to confer ganciclovir resistance, emerged rapidly in a heart-transplant recipient after 54 days of antigenaemia-guided ganciclovir therapy. The emergence of this viral mutant, while the patient was receiving oral ganciclovir 1000 mg three times daily, was associated with increasing CMV pp65 antigenaemia levels despite the re-instatement of intravenous ganciclovir 5 mg/kg twice daily. The antiviral regimen was then switched to intravenous foscarnet 90 mg/kg, administered either twice daily, once daily or every other day, which resulted in a rapid decrease of CMV antigenaemia levels and prevented the development of CMV-associated disease. Frequent monitoring of the CMV viral load and/or mutational studies are warranted in highly immunocompromised transplant recipients to prevent the development of CMV disease caused by ganciclovir resistant strains. PMID- 11867026 TI - Immunobiology of herpes simplex virus (HSV) infections. PMID- 11867027 TI - Immunity to herpes simplex virus: a hypothesis. AB - This article discusses some possible reasons why people vary in susceptibility to primary infection by herpes simplex virus (HSV), and, in addition, why some individuals suffer lesions upon viral reactivation from latency. An underlying hypothesis is that the effectiveness of various aspects of innate immunity, conditioned by the person's microbial exposure in early life, may affect susceptibility to HSV infection, the nature of the initial adaptive immune response, and the efficacy of the memory-recall reaction following reactivation. Could domicile in a super-sanitized environment result in more troublesome problems with HSV such as is suspected to explain the rising tide of allergy? PMID- 11867028 TI - The Holy Grail: immune control of human herpes simplex virus infection and disease. AB - To develop an effective vaccine against genital herpes, the mechanisms of immune control of primary and recurrent genital herpes need to be elucidated thoroughly in humans, using animal models as a guide. The types of effector cells and their main viral target/stimulator proteins are especially important. In human recurrent herpes simplex virus (HSV) infections, the major effectors are CD4 and CD8 lymphocytes and T-helper 1 cytokines, interferon-in particular. Glycoprotein D, B2-tegument proteins and proteins produced early in viral replication are stimulatory for CD4 and CD8 lymphocytes, respectively. These in vitro results are consistent with the findings of the most recent HSV vaccine trials. PMID- 11867029 TI - Acute coronary syndrome is a common clinical presentation of in-stent restenosis. AB - Coronary stents have been the major advancement in percutaneous coronary intervention in the last decade and are used in 60% to 80% of patients. However, in-stent restenosis continues to be a problem, occurring in 20% to 30% of cases. The clinical presentation of patients who develop restenosis after stenting has not been well characterized. In this study we compared the clinical presentation of in-stent restenosis with that of restenosis without stenting. Of 739 patients who underwent percutaneous coronary intervention and had repeat catheterization between October 1, 1997, and June 30, 2000, 262 consecutive patients with recurrent ischemia and restenosis were identified: 191 patients with (group A) and 71 without (Group B) stenting. Patients who underwent interventions in bypass grafts and those who developed early acute stent thrombosis were excluded from the study. Recurrent clinical ischemia occurred at a mean of 5.5 months in group A and 6.5 months in group B (p = 0.24). Rest angina (Braunwald class II and III) was more frequent in group A (48% vs 32%, p = 0.032). Acute coronary syndromes, the combination of rest angina, and acute myocardial infarction were also more frequent in group A (68% vs 46%, p = 0.03). Patients in group A were more likely to have angiographically visible thrombus than those in group B (9% vs 0%, p = 0.02). Thus, acute coronary syndromes are a common clinical presentation of restenosis among patients whose follow-up angiogram is obtained for clinical reasons, and occur more frequently in patients with in-stent restenosis than in those with restenosis without stenting. PMID- 11867030 TI - Relation of homocysteine, vitamin B(12), and folate to coronary in-stent restenosis. AB - Coronary in-stent restenosis represents a clinical problem. Because homocysteine is being discussed as a new risk factor for atherosclerosis and thrombosis, this study investigated the relations of homocysteine, folate, and vitamin B(12) to the rate of in-stent restenosis. Patients undergoing successful percutaneous transluminal coronary angioplasty of native coronary lesions with stent implantation were investigated for fasting total serum homocysteine, folic acid, and vitamin B(12). The rate of in-stent restenosis was determined angiographically after 6 months, or earlier if clinically indicated. Of 292 enrolled patients, 262 (90%) (189 men and 73 women) underwent control angiography on an average of 6.3 +/- 1.0 (SD) months after intervention. The rate of in-stent restenosis was 36%. Univariate and multivariate analyses revealed no significant differences between patients with or without restenosis with regard to total homocysteine (median [interquartile range]: 12.9 [11.2 to 14.8] and 12.4 [10.3 to 15.4] micromol/L, respectively), folate (16.1 [12.4 to 20.5] and 15.4 [12.5 to 19.5] nmol/L, respectively), or vitamin B(12) (239.0 [182.5 to 322.1] and 258.4 [205.8 to 330.5] pmol/L, respectively). These results suggest that homocysteine, folate, and vitamin B(12) are not related to the angiographically determined rate of coronary in-stent restenosis after 6 months. PMID- 11867031 TI - Chest pain after coronary artery stent implantation. AB - A sizeable proportion of patients who undergo successful coronary artery stent implantation experiences chest pain immediately after the procedure and/or in the following months in the absence of in-stent restenosis. We investigated this phenomenon in 57 consecutive patients with stable angina who underwent successful stent implantation. Chest pain characteristics were assessed before stent implantation and during 6-month follow-up. All patients underwent coronary angiography within 6 months of the procedure 48 hours after exercise thallium-201 perfusion scintigraphy. Patients who did not exhibit in-stent restenosis underwent an ergonovine test at the end of routine coronary angiography. During follow-up, 15 patients complained of chest pain. Six of these patients exhibited scintigraphic evidence of myocardial ischemia and in-stent restenosis at angiography. In the remaining 9 patients, chest pain occurred in the absence of in-stent restenosis at angiography. In 8 of these patients intracoronary ergonovine administration reproduced their habitual pain, whereas it did not cause any pain in the 42 patients who were completely asymptomatic at follow-up and without in-stent restenosis. Ergonovine caused more intense vasoconstriction and nitroglycerin caused more intense vasodilation of the reference coronary diameter in patients with than in patients without ergonovine-induced pain (-17 +/- 3 vs -9 +/- 3%, p <0.001; 9 +/- 6 vs 5 +/- 4%, p <0.02, respectively). In conclusion, chest pain with features similar to habitual angina occurs in the absence of in-stent restenosis in 1/5 of patients after stent implantation and appears to be associated with more intense coronary vasoreactivity. PMID- 11867032 TI - Mechanisms of acute gain and late lumen loss after atherectomy in different preintervention arterial remodeling patterns. AB - The main mechanism of restenosis after directional coronary atherectomy (DCA) remains obscure. We investigated mechanisms of restenosis after DCA in different coronary artery remodeling patterns. DCA was performed in 51 de novo lesions. The lesions were evaluated by intravascular ultrasound (IVUS) before, immediately after, and 6 months after the procedure. According to the IVUS findings before DCA, we classified the lesions into the following 3 groups: (1) positive (n = 10), (2) intermediate (n = 25), and (3) negative (n = 16) remodeling. We measured lumen area, vessel area, and plaque area using IVUS before DCA, immediately after DCA, and at follow-up. Lumen area increase after DCA was mainly due to plaque area reduction in the positive and intermediate remodeling groups (90 plus minus 15% and 80 plus minus 25% increase in lumen area, respectively), whereas that in the negative remodeling group was due to both plaque area reduction (57 plus minus 22% increase in lumen area) and vessel area enlargement (43 plus minus 33% increase in lumen area). The plaque area increase correlated strongly with late lumen area loss in the positive and intermediate remodeling groups (r = 0.884, p <0.001; r = 0.626, p <0.001, respectively), but the decrease in vessel area was not correlated with lumen area loss. In contrast, both an increase in plaque area and a decrease in vessel area were correlated with late lumen area loss (r = 0.632, p = 0.009; r = 0.515, p = 0.041) in the negative remodeling group. Coronary artery restenosis after atherectomy was primarily due to an increase in plaque in the positive and/or intermediate remodeling groups. However, in the negative remodeling group, late lumen loss might have been caused by both an increase in plaque and vessel shrinkage. PMID- 11867033 TI - Gender differences in acute myocardial infarction in the era of reperfusion (the MITRA registry). AB - There is conflicting information about gender differences in presentation, treatment, and outcome after acute ST elevation myocardial infarction (STEMI) in the era of thrombolytic therapy and primary percutaneous coronary intervention. From June 1994 to January 1997, we enrolled 6,067 consecutive patients with STEMI admitted to 54 hospitals in southwest Germany in the Maximal Individual TheRapy of Acute myocardial infarction (MITRA), a community-based registry. Women were 9 years older than men, more often had hypertension, diabetes mellitus, and congestive heart failure, and had a history of previous myocardial infarction less often. Women had a longer prehospital delay (45 minutes), had anterior wall infarction more often (odds ratio [OR] 1.21; 95% confidence interval [CI] 1.08 to 1.36), and received reperfusion therapy less often (OR 0.83; 95% CI 0.74 to 0.94). The percentage of patients who were eligible for thrombolysis and received no reperfusion was higher in women (OR 1.7; 95% CI 1.56 to 1.89). Women had recurrent angina (OR 1.45; 95% CI 1.23 to 1.71) and congestive heart failure (OR 1.26; 95% CI 1.01 to 1.56) more often. There was a trend toward a higher hospital mortality in women (age-adjusted OR 1.16, 95% CI 0.99 to 1.35; multivariate OR 1.21, 95% CI 0.96 to 1.51), but there was no gender difference in long-term mortality after multivariate analysis (age-adjusted OR 0.95, 95% CI 0.78 to 1.15; multivariate OR 0.93, 95% CI 0.72 to 1.19). Thus, women with STEMI receive reperfusion therapy less often than men. They experience recurrent angina and congestive heart failure more often during their hospital stay. The age-adjusted long-term mortality is not different between men and women, but there is a trend for a higher short-term mortality in women. PMID- 11867034 TI - Prognostic value of left atrial enlargement in patients with idiopathic dilated cardiomyopathy and ischemic cardiomyopathy. AB - Previous studies have shown that abnormal mitral flow patterns and left atrial (LA) enlargement are independently associated with survival in patients with left ventricular (LV) dysfunction. However, it is not known whether these outcome indicators can provide different information in patients of various age groups. This study was designed to assess the prognostic value of the restrictive mitral flow pattern (RMFP) and increased LA size in patients with LV dysfunction (ejection fraction <45%) grouped into those < or = 70 years old (n = 102; mean age 61) and those >70 years old (n = 105; mean age 78). Echocardiographic and Doppler indexes were recorded in patients with LV systolic dysfunction due to dilated cardiomyopathy who were followed up for 22 plus minus 14 months. In patients >70 years, indexed LA size (>26 mm/m(2)) was the single best predictor of death (hazard ratio [HR] 3.0, p = 0.018) and emerged as the most important outcome variable of the combined end point (HR 2.2, p = 0.016) on multivariate analysis. In patients < or =70 years, RMFP, characterized by an early wave deceleration time <140 ms, was independently associated with cardiac death or heart failure hospitalization (HR 5.7, p = 0.0013). When demographics, clinical, echocardiographic, and Doppler measurements were analyzed in hierarchical order, indexed LA size yielded the most valuable contribution in predicting the combined end point in older patients (global chi-square from 11.5 to 18.7). RMFP was associated with the higher additional prognostic value in younger patients (global chi-square from 14.4 to 24.1). These data suggest that LA enlargement has an independent and additional prognostic value in elderly patients with LV dysfunction. PMID- 11867035 TI - Cardiovascular risk factors in ethnic minority women aged < or =30 years. AB - Men and women of African and South Asian ancestry in the United States are increasingly recognized as being at greater risk for coronary heart disease (CHD) than Caucasians of European ancestry. Relatively little data on the genetic and lifestyle risk factors that predispose women to CHD in these ethnic minorities are available. We compared coronary risk factors in a volunteer sample of African American, Asian Indian American, and Caucasian American women of college age. Life style, dietary, hemodynamic and anthropometric parameters, and laboratory data were sought from 70 subjects in each ethnic group. African-American women were found to have lower triglyceride levels and higher apolipoproten A-1, high density lipoprotein (HDL), lipoprotein (a) (Lp(a)), fibrinogen, and fasting insulin levels. They also consumed more fat and cholesterol than their peers, had a higher percentage of body fat, body weight, and body mass indexes, and reported less physical activity than Caucasians. Asian Indian American women had higher Lp(a), HDL, and fibrinogen levels than Caucasian American women, and also reported less physical activity. Thus, young African- American and Asian Indian American women have several modifiable risk factors as well as some nontraditional lipid risk factors that warrant consideration for explaining the increased prevalence of CHD in these ethnic groups. The tendency toward peripheral insulin insensitivity and increased body fat in this age group of African-American women suggests diet and exercise may reduce the risk of subsequent CHD. PMID- 11867036 TI - Prevalence and prognostic significance of psychiatric disorders in patients evaluated for recurrent unexplained syncope. AB - We aimed to assess the psychiatric profile and prognostic value of psychiatric disorders (PDs) in patients presenting with unexplained syncope. Forty patients with recurrent unexplained syncope referred for head-up tilt testing were compared with age- and sex-matched patients free of known chronic PDs referred for arrhythmia. All patients underwent a semistandardized psychiatry questionnaire (Mini-International Neuropsychiatric Interview) to assess their profile. Additional stress coping was performed to study adaptational processes to stressful situations. After tilt testing and psychiatric evaluation, a drug free follow-up was performed in patients with syncope. Of the 80 patients who referred to the psychiatric interview, 40 had evidence of at least 1 psychiatric disorder. They were 26 patients (65%) in the syncope group and 14 patients (35%) in the control group (p = 0.01). Detailed analysis revealed a more frequent subprofile of anxiety and panic disorders in patients with syncope than in controls (30% vs 12% and 20% vs 10%, respectively), whereas the subprofile of depression was similar in both groups. Moreover, those with syncope were more likely to have a high anxiety index (25 +/- 5 vs 22 +/- 4, p = 0.004), and were more prone to avoidance-oriented coping strategies when experiencing undesirable life events than controls. Considering syncope patients, no difference could be found between the 25 with a positive tilt test and the 15 with a negative tilt test with respect to the number of syncopal episodes and psychiatric profile. After a 3-year drug-free follow-up, 15 patients (37.5%) had at least 1 recurrent syncope. The recurrence rate was similar in patients with positive and negative head-up tilt test results (9 of 25 vs 6 of 15, respectively). In contrast, the syncopal recurrence rate was higher in patients who fulfilled criteria for affective disorders (13 of 26 vs 2 of 14, 95% confidence interval 1.09 to 2.55, relative risk 1.7, p = 0.04). Thus, patients with recurrent unexplained syncope are more anxious and are more prone to panic disorders and avoidance-oriented coping strategies than control patients with arrhythmia. The presence of a psychiatric disorder is associated with an increased risk of recurrence. The outcome of such patients may be improved with recognition and treatment of PDs. PMID- 11867037 TI - Blood pressure responses to acute stress and left ventricular mass (The Hypertension Genetic Epidemiology Network Study). AB - Left ventricular (LV) hypertrophy is a major risk factor for cardiovascular morbidity. This study examined the extent to which exaggerated blood pressure (BP) reactivity is associated with LV mass and relative wall thickness (RWT) in a large sample of middle-aged hypertensive men and women. Participants included 1,012 African-Americans and 655 Caucasians recruited for the Hypertension Genetics Epidemiology Network (HyperGEN) study. A laboratory stress protocol was implemented, consisting of BP measurement during rest, handgrip, and a mental arithmetic stressor. This was followed by a 2-dimensional guided M-mode echocardiographic evaluation to measure LV mass. LV mass was greater among African-Americans relative to Caucasians (p <0.01). Systolic BP at rest was positively associated with LV mass and RWT in African-Americans (p <0.001) and in Caucasian women (p <0.0001). LV mass and RWT were compared in participants and participants were classified as high or low reactors based on their systolic BP responses to the 2 laboratory stressors. RWT demonstrated positive associations with systolic BP responses to both tasks. Associations with LV mass were mostly nonsignificant with the exception of a negative association in African-Americans between systolic BP responses to the arithmetic task and LV mass. We conclude that in this cohort of hypertensive middle-aged men and women, BP reactivity did not predict LV mass. Positive associations of systolic BP responses with RWT suggest that exaggerated reactivity may particularly be related to LV changes influenced by vascular tonus. PMID- 11867038 TI - Long-term follow-up of patients after coarctation of the aorta repair. AB - Late cardiovascular complications after operative repair of coarctation of the aorta include systemic hypertension, premature coronary artery disease, aortic valve abnormalities, aortic aneurysm, and recoarctation. We report the outcome in 274 subjects greater-than-or-equal50 years after coarctation repair. Operative repair of simple coarctation was performed on 274 patients at the University of Minnesota Hospital between 1948 and 1976. Twenty patients (7%) died in the immediate postoperative period. Of the 254 survivors, 2 were lost to follow-up, 45 (18%) died at a mean age of 34 years, and 207 (81%) were alive greater-than-or equal50 years after the original operation. Coronary artery disease and perioperative deaths at the time of a second cardiac operation accounted for 17 of the 45 late deaths. Predictors of survival were age at operation and blood pressure at the first postoperative visit. Of the 207 long-term survivors, 92 (48%) participated in a clinical cardiovascular evaluation. Thirty-two of the 92 subjects had systemic hypertension that was predicted by age at operation, blood pressure at the first postoperative visit, and paradoxic hypertension at operative repair. New cardiovascular abnormalities detected at follow-up evaluation included evidence of a previous myocardial infarction, cardiomyopathy, atrial fibrillation, moderate to severe left ventricular outflow tract obstruction, moderate aortic valve regurgitation, recoarctation, and ascending aortic dilation. Thus, long-term survival is significantly affected by age at operation, with the lowest mortality rates observed in patients who underwent surgery between 1 and 5 years of age. More than 1/3 of the survivors developed significant late cardiovascular abnormalities. PMID- 11867039 TI - Clinical significance of echo-free space detected by transesophageal echocardiography in patients with type B aortic intramural hematoma. AB - To evaluate the prevalence and clinical significance of echo-free space (EFS) in aortic intramural hematoma (AIH) during transesophageal echocardiography (TEE), TEE performed during the acute phase in 71 consecutive patients with type B AIH was reviewed. Forty-four patients (62%) had EFS including 24 patients with a large EFS occupying >1/2 of the hematoma thickness. Among 59 patients who also underwent computed tomography, focal contrast enhancement in the hematoma area was observed in only 7 patients with a large EFS. Hospital mortality and incidence of surgical intervention in patients with EFS were 0% and 2%, respectively, which was similar to 4% in those without EFS. Follow-up imaging studies in 57 patients (80%) revealed development of typical aortic dissection (AD) in 6 patients and complete resorption of hematoma in 35; the incidence of either the development of AD or a complete resorption of hematoma was not significantly different between those with and without EFS. EFS by TEE is not rare in patients with type B AIH. It is not a poor prognostic factor and is not associated with the development of AD. PMID- 11867040 TI - Analysis of left atrial volume change rate during left ventricular diastolic phase with M-mode echocardiography for differentiation between normal and pseudonormal mitral inflow. AB - Pseudonormalization of mitral inflow is a diagnostic problem in clinical practice. An excellent correlation exists between the change in the left atrial (LA) angiographic area and posterior aortic wall motion. Therefore, we sought to define the role of LA wall motion, indicating LA volume change rate, in patients with normal and pseudonormal mitral inflow. We performed echocardiography after cardiac catheterization in 62 patients with a velocity ratio of early-to-late mitral inflow (E/A ratio) >1. Study patients were classified into 2 groups according to the response of mitral inflow to phase II of the Valsalva maneuver: patients with E/A >1 after the Valsalva maneuver (n = 31, control group), and patients with <1 after the Valsalva maneuver (n = 31, pseudonormal group). The slopes (slope E and A) of early diastolic and late diastolic motion of the LA wall were derived from M-mode analysis together with the time constant of left ventricular (LV) isovolumic relaxation from cardiac catheterization. The values of slope E (41 plus minus 11 vs 61 plus minus 12 mm/s, p <0.001) and slope E/A (0.69 plus minus 0.13 vs 1.32 plus minus 0.35, p <0.001) were significantly lower in the pseudonormal group and were inversely correlated with the time constant of LV isovolumic relaxation (r = 0.64, p <0.001 and r = 0.73, p <0.001, respectively). Using slope E/A <1 as an indicator of relaxation abnormality, the sensitivity, specificity, positive predictive value, and negative predictive value for the detection of pseudonormalization were 94%, 100%, 100%, and 94%, respectively. The slope of LA wall motion, indicating LA volume change rate, during the LV diastolic phase is useful for evaluating pseudonormal LV diastolic dysfunction in the selected patient population. PMID- 11867041 TI - Effects of rapid saline infusion on orthostatic intolerance and autonomic tone after 20 days bed rest. AB - To test whether acute volume expansion can normalize orthostatic intolerance and autonomic tone after prolonged bed rest (BR), 23 men were subjected to 20 days BR. Left ventricular (LV) echocardiography was performed during the lower body negative pressure (LBNP) test before and after BR with and without preceding rapid infusion of saline (1,500 ml/30 min). Saline infusion restored heart rate, LV dimension, and stroke volume during LBNP, increased cardiac output (from 4.1 +/- 1 to 5.3 +/- 1 L/min), and normalized LBNP tolerance time (from 11 +/- 4 to 23 +/- 6 minutes). In 9 men, a Holter electrocardiogram was recorded on the day before BR, the fourth and twentieth days of BR, and the day after BR. The high frequency component of heart rate variability during sleep gradually decreased and reached the lowest level on the day after BR (100%, 66 +/- 16%, 39 +/- 18%, 10 +/- 8%). Thus, restoring decreased blood volume is an effective countermeasure for orthostatic intolerance after BR. However, decreased vagal tone persisted, suggesting reset autonomic tone. PMID- 11867044 TI - Joseph Kayle Perloff, MD: a conversation with the editor. PMID- 11867042 TI - Effect of losartan in aging-related endothelial impairment. AB - Aging is associated with progressive deterioration in endothelial function. We hypothesized that losartan may represent a useful therapeutic strategy to ameliorate endothelial function in aged subjects. Eighteen healthy older subjects (mean age 75 +/- 3 years) were prospectively randomized in a double-blind, crossover fashion to receive either losartan 50 mg/day or placebo for 6 weeks. Subjects were switched to the opposite arm after a 2- week washout period. Flow mediated dilation (FMD) in the brachial artery and plasma levels of vascular cell adhesion molecule-1, intercellular adhesion molecule (ICAM), moncocyte chemoattractant 1 protein, and E-selectin were measured in both arms at the beginning and end of the 6-week period. Losartan resulted in a 6-mm Hg decrease in systolic blood pressure (from 130 +/- 12 to 124 +/- 13 mm Hg), which was no different from placebo (132 +/- 12 to 127 +/- 13 mm Hg). FMD increased from 3.1 +/- 0.6% to 3.9 +/- 0.6% after losartan, and decreased from 3.3 +/- 0.3% to 2.4 +/- 0.6% after placebo (p = NS for both). In contrast, losartan reduced circulating concentrations of vascular cell adhesion molecule 1 (750 +/- 73 to 572 +/- 39), ICAM (405 +/- 26 to 196 +/- 10), and moncocyte chemoattractant 1 protein (560 +/- 56 to 423 +/- 35) (p <0.01 for all by analysis of variance), but not E-selectin. On univariate analyses, the strongest predictor of baseline endothelial function and change in FMD with losartan was low-density lipoprotein. There was a negative correlation between baseline endothelial function and change in FMD in response to losartan (r(2) = -0.75, p = 0.0003). Baseline ICAM levels alone significantly correlated with low-density lipoprotein cholesterol (r(2) = 0.54, p = 0.02) and weakly correlated with total cholesterol (r(2) = 0.47, p = 0.05). Thus, administration of losartan for a duration of 6 weeks has favorable effects on inflammatory markers in healthy older subjects, but does not alter peripheral conduit endothelial function. PMID- 11867043 TI - Need for a moratorium on percutaneous transluminal coronary angioplasty in stable coronary artery disease. PMID- 11867045 TI - Postprocedure creatine kinase-MB elevation and baseline left ventricular dysfunction predict one-year mortality after percutaneous coronary intervention. PMID- 11867046 TI - Comparison of effects of dalteparin and enoxaparin on hemostatic parameters and von Willebrand factor in patients with unstable angina pectoris or non--ST- segment elevation acute myocardial infarction. PMID- 11867047 TI - Diastolic dysfunction, infarct size, and exercise capacity in remote myocardial infarction: a combined approach of mitral E-wave deceleration time and color M mode flow propagation velocity. PMID- 11867048 TI - Blood glucose concentrations < or = 125 mg/dl and coronary heart disease risk. PMID- 11867049 TI - Results and complications of the carotid sinus massage performed according to the "method of symptoms". PMID- 11867050 TI - Long-term outcome of painful left bundle branch block. PMID- 11867051 TI - Relation between neurohormonal activation and enhanced ventilatory response to exercise in patients with chronic congestive heart failure. PMID- 11867052 TI - Effects of percutaneous mitral balloon valvuloplasty on P-wave dispersion in patients with mitral stenosis. PMID- 11867053 TI - Usefulness of Doppler tissue imaging analysis of tricuspid annular motion for determination of right ventricular function in normal infants and children. PMID- 11867054 TI - Bedside balloon atrial septostomy is safe, efficacious, and cost-effective compared with septostomy performed in the cardiac catheterization laboratory. PMID- 11867055 TI - Gender comparisons in pulmonary embolism (results from the International Cooperative Pulmonary Embolism Registry [ICOPER]). PMID- 11867056 TI - Circadian, weekly, and seasonal variation at the onset of acute aortic dissection. PMID- 11867057 TI - Iatrogenic aortic dissection. PMID- 11867058 TI - Paradoxical inferior-posterior wall systolic expansion in patients with end-stage liver disease. PMID- 11867060 TI - Aortic valve calcium on spiral computed tomography is associated with calcification of the thoracic aorta in hypertensive patients. PMID- 11867059 TI - Association between n-3 fatty acid status in blood and electrocardiographic predictors of arrhythmia risk in healthy volunteers. PMID- 11867061 TI - Intracardiac echocardiography in the interventional catheterization laboratory: preliminary experience with a novel, phased-array transducer. PMID- 11867062 TI - Assessment of myocardial perfusion during pharmacologic contrast stress echocardiography. PMID- 11867063 TI - Effect of early administration of atropine on paradoxic sinus deceleration during dobutamine stress echocardiography. PMID- 11867065 TI - Double counting of acute myocardial infarction makes estimates of occurrence and case fatality inaccurate. PMID- 11867064 TI - Suboptimal early inhibition of platelets by treatment with tirofiban and implications for coronary interventions. PMID- 11867066 TI - Cardioprotective effects of preinfarction angina in elderly: same patients, different results? PMID- 11867067 TI - Correlation of aortic stiffness and restenosis after coronary angioplasty. PMID- 11867068 TI - Determination of cadmium and zinc in Alzheimer's brain tissue using inductively coupled plasma mass spectrometry. AB - In this work, brain tissue was taken from Alzheimer's Disease (AD) subjects (n=11), 'normal' subjects (n=10) and from subjects with senile involutive cortical changes (SICC) (n=6). Concentrations of Cd and Zn were determined in all samples, using Inductively Coupled Plasma Mass Spectrometry (ICP-MS). The brain tissue was selected and obtained from the Netherlands Brain Bank. Samples were taken in each case, from both hemispheres of the superior frontal gyrus, the superior parietal gyrus, the medial temporal gyrus, the hippocampus and the thalamus of the same brain.Cd which is known to have no essential role in the brain was found to follow, as expected, a lognormal distribution of concentrations in 'normal' subjects (Shapiro-Wilk's test (0.98) (p<0.18)). For the Alzheimer's Disease subjects and SICC subjects, the data tends to follow a lognormal distribution, rather than a normal distribution, but is still significantly different from it (Shapiro-Wilk's test (0.97) (p<0.03); (0.93) (p<0.0067), respectively)). In the case of Zn concentrations, the data tends to follow a normal distribution for the 'normal' subject group, even though the data is significantly different from it (Shapiro-Wilk's test (0.95) (p<0.001)). Whereas in the Alzheimer's Disease and SICC subject groups, the data follows a normal distribution (Shapiro-Wilk's test (0.98) (p<0.21); (0.97) (p<0.2002), respectively)). When comparing age-matched groups, for all regions and both hemispheres, no significant differences (p>0.1) for Cd were found between 'normals' and Alzheimer's Disease subjects and Alzheimer's Disease subjects and SICC but at a low level of significance, lower concentrations of Cd were found in the SICC group compared to the 'normals'. For all regions and both hemispheres, Zn was found to be significantly decreased in the Alzheimer's Disease group, compared to the 'normal' and SICC groups. Zn concentrations were also found to be significantly decreased in the 'normals' compared to the SICC group. It is also of interest that Cd negatively correlates with the scale of tangles in both 'normals' (p<0.001) and Alzheimer's Disease subjects (p<0.01). In the SICC subjects Cd correlates negatively with the tangles but not significantly so (p>0.1). PMID- 11867070 TI - Frontal lobe dysfunction in amyotrophic lateral sclerosis. AB - The aim of the present study was to investigate the involvement of frontal lobe dysfunction in amyotrophic lateral sclerosis (ALS) using ocular motor paradigms and neuropsychological testing. Fifty-one patients with ALS participated in the following ocular motor tasks: (1) a three-choice task and (2) a remembered saccade task. The patients underwent a clinical and neuropsychological evaluation. One-third of ALS patients presented with signs of frontal dysfunction, as determined by their high distractibility factors (DF) in the three-choice task and their performances in both the Wisconsin and Stroop tests. ALS patients exhibited longer latencies to eye movement than controls in the performance of the remembered saccade task, specifically in performance of both remembered and delayed saccades, but saccade accuracy was not impaired. Finally, performance indices of the ocular motor tasks, in particular the DF, was correlated only with the degree of dysarthria. PMID- 11867071 TI - Macrophage infiltration and death in the nerve during the early phases of experimental diabetic neuropathy: a process concomitant with endoneurial induction of IL-1beta and p75NTR. AB - This study describes the infiltration and death of monocyte/macrophages and concomitant endoneurial expression of the pro-inflammatory cytokine interleukin 1beta (IL-1beta) and neurotrophin receptor p75 (p75NTR) in the sciatic nerve at the early phases of experimental diabetic neuropathy induced in Lewis rats by streptozotocin (STZ) intraperitoneal injection. Immunocytochemistry and single nerve fiber immunostaining showed the presence of macrophages in diabetic nerves by weeks 2 and 3 after STZ administration, and the 15% of these cells were TUNEL positive. IL-1beta was evident in scattered macrophages, and along few isolated nerve fibers until week 5, when it became undetectable, in concomitance with complete endoneurial clearance of macrophages. p75NTR showed an up-regulation in the sciatic nerve of diabetic rats that began by week 3 after STZ administration, reached its peak by week 5, and returned then to a barely detectable level by week 6. These findings seem to indicate that macrophages and IL-1beta may be involved in the pathogenesis of diabetic neuropathy, participating not only to nerve damage but also to the promotion of an attempt of regeneration via p75NTR induction. PMID- 11867072 TI - IP-10 and MCP-1 levels in CSF and serum from multiple sclerosis patients with different clinical subtypes of the disease. AB - Interferon-gamma-inducible Protein-10 (IP-10) and Monocyte Chemotactic Protein-1 (MCP-1) levels were measured by enzyme-linked immunosorbent assay (ELISA) in the CSF and in the serum from 74 patients affected by different clinical forms of Multiple Sclerosis (MS), including 39 patients with Relapsing Remitting (RR) MS in an active phase, 14 patients in a stable phase of the disease, 12 patients with Secondary Progressive (SP) MS and 9 patients with Primary Progressive (PP) MS. IP-10 and MCP-1 levels were also determined in 19 subjects with no neurological diseases or major systemic disorders, 18 patients with non inflammatory neurological diseases, as well as in 15 patients with other inflammatory neurological diseases.IP-10 levels were significantly elevated in CSF and serum from RR and SP, but not PP-MS patients. On the contrary, MCP-1 levels were decreased in CSF and serum of all MS patients. CSF concentrations of IP-10 and MCP-1 did not significantly correlate neither with each other, nor with CSF mononuclear cell count, albumin quotient or CSF IgG index. No correlation between disease duration, clinical course or EDSS score and chemokine levels was found.IP-10 and MCP-1 undergo modifications in different subtypes of the disease: IP-10 levels in CSF and serum samples are markedly increased when inflammation is prominent, and not in PP--MS patients, where inflammation is less evident. MCP-1 decrease in CSF and serum from MS patients could be related to the regulation of T-cell polarization. PMID- 11867069 TI - Abnormal palatopharyngeal muscle morphology in sleep-disordered breathing. AB - The aim of the present study was to investigate whether histopathological changes can be detected in two soft palate muscles, the palatopharyngeus and the uvula, in 11 patients with long duration of sleep-disordered breathing (SDB). Muscle samples were collected from patients undergoing uvulo-palatopharyngoplasty (UPPP). Reference samples from the corresponding areas were obtained at autopsy from five previously healthy subjects. Muscle morphology, fibre type and myosin heavy chain (MyHC) compositions were analysed with enzyme-histochemical, immunohistochemical and biochemical techniques. The muscle samples from the patients, and especially those from the palatopharyngeus, showed several morphological abnormalities. The most striking findings were (i) increased amount of connective tissue, (ii) abnormal variability in fibre size, (iii) increased proportion of small-sized fibres, (iv) alterations in fibre type and MyHC compositions, (v) increased frequency of fibres containing developmental MyHC isoforms. Our findings point towards a pathological process of denervation and degeneration in the patient samples. Conclusively, the morphological abnormalities suggest a neuromuscular disorder of the soft palate in SDB patients. PMID- 11867073 TI - Castleman's disease-associated neuropathy: no evidence of human herpesvirus type 8 infection. AB - Castleman's disease (CD) is a rare lymphoproliferative disorder that may be associated with a neuropathy. In a recent report, the presence of human herpesvirus type 8 (HHV-8) DNA sequences were detected in an HIV-negative patient with polyneuropathy, organomegaly, endocrinopathy, M-protein and skin changes (POEMS) associated with the multicentric hyaline vascular variant of CD. It was proposed that the presence of these sequences may have a role in the pathophysiology of the neuropathy. We describe an HIV-negative woman with the multicentric plasma cell form of CD who presented with a disabling neuropathy. In addition to a severe demyelinating polyneuropathy, she had some of the other features of POEMS including an IgA lambda gammopathy and lymphadenopathy. We were unable to detect the presence of either HHV-8 DNA or proteins in this patient. The significance of our results and the relationship between CD, POEMS and neuropathy are discussed. PMID- 11867074 TI - Cytokine regulation of E-selectin in rat CNS microvascular endothelial cells: differential response of CNS and non-CNS vessels. AB - We have compared the induced expression of E-selectin in primary cultures of rat brain microvascular endothelial cells (EC), pericytes and in non-CNS microvascular endothelium stimulated with the cytokines, IL-1beta (20 ng/ml), and tumor necrosis factor (TNF)-alpha (75 ng/ml). Expression was studied at both the protein and mRNA levels. Fluorescence in-situ hybridization (FISH) was used to examine de novo synthesis of E-selectin mRNA. Laser cytometric analysis was used as a novel approach to the quantitaion of FISH. In-situ hybridization was performed using two PCR-generated probes. The first probe (517 bp) spanned the lectin and epidermal growth factor (EGF)-like domain. The second probe (562 bp) spanned the CR3, 4, and 6 domains. E-selectin-specific mRNA was localized to the perinuclear regions of the EC. Both cytokines, IL-1beta and TNF-alpha significantly increased E-selectin gene expression in CNS EC but not pericytes. IL-1beta induced higher E-selectin mRNA levels than TNF-alpha. The maximum number of mRNA-positive cells was observed after stimulation for 4--6 h. Surface protein expression was sustained for up to 48 h following addition of cytokine. This was in contrast to the transient expression in non-CNS EC indicating that pure primary CNS EC display slightly different kinetics of E-selectin expression than non-CNS EC. PMID- 11867075 TI - ABO-incompatible auxiliary partial orthotopic liver transplant for late-onset familial amyloid polyneuropathy. AB - A 60-year-old Japanese man with late-onset familial amyloid polyneuropathy type I (FAP transthyretin Met30) showed clinical improvement following auxiliary partial orthotopic liver transplantation (APOLT) from an ABO-incompatible living related donor. Preoperatively, plasmapheresis and immunosuppressant drugs were used to reduce serum antibodies against the donor's ABO type. APOLT was chosen so the residual liver could sustain the patient in the event of hyperacute rejection. OLT is applicable to late-onset FAP transthyretin Met30, and APOLT can be considered in ABO-incompatible cases. PMID- 11867076 TI - The serum level of free testosterone is reduced in amyotrophic lateral sclerosis. AB - Sporadic amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disorder affecting upper and lower motoneurons. There is an approximately 2:1 higher incidence of ALS in men compared to women, and this has raised the hypothesis of an involvement of sex hormones in the etiopathogenesis of the disorder. In this work, the serum levels of dehydroepiandrosterone sulphate (DHEAS), 17-betaestradiol, free and total testosterone were measured in 35 patients with defined or probable ALS, according to the El-Escorial/WFN revisited criteria, and compared to those obtained from 57 disease controls, matched for age and gender to the ALS group. We found no differences between ALS cases and disease controls in the serum levels of DHEAS, 17-betaestradiol and total testosterone. Conversely, free testosterone was significantly decreased in the ALS group. Given that testosterone crosses the blood-brain barrier only as unbound form, we suggest a possible involvement of this sex hormone in the pathophysiology of this severe motor neuron disease. PMID- 11867077 TI - Autosomal dominant palatal myoclonus and spinal cord atrophy. AB - We report a new family with palatal myoclonus, pyramidal tract signs, cerebellar signs, marked atrophy of the medulla oblongata and spinal cord, and autosomal dominant inheritance. These findings were almost identical with those in patients previously reported to have histopathologically confirmed adult-onset Alexander disease. Recently, heterozygous point mutations in the coding region of glial fibrillary acidic protein (GFAP) in patients with an infantile form of Alexander disease have been reported. We found a new heterozygous amino acid substitution, Val87Gly in exon 1 of GFAP, in the affected individuals in this family but not in 100 spinocerebellar ataxia (SCA) patients and 100 controls. Therefore, this family might have new clinical entities related to adult-onset Alexander disease and GFAP mutation. PMID- 11867078 TI - Sympathetic and parasympathetic pupillary dysfunction in familial dysautonomia. AB - Objective assessment of autonomic dysfunction in familial dysautonomia (FD) is largely based on the analysis of cardiovascular responses to challenge maneuvers such as orthostatic stress. Infrared pupillometry (IPM) provides an additional reliable method for cranial autonomic evaluation and has the advantage of requiring minimal cooperation.This study was performed to determine whether IPM contributes to the assessment of autonomic function in FD patients. In 14 FD patients and 14 healthy controls, we studied absolute and relative light reflex amplitude, pupillary constriction velocity (v(constr)), pupillary diameter, early and late pupillary re-dilatation velocity (v(dil 1), v(dil 2)) after dark adaptation. Prior to IPM, all patients had an ophthamological examination to evaluate refraction and corneal integrity. In comparison to controls, patients had a significant reduction of the parameters reflecting parasympathetic pupillary function (absolute light reflex amplitude 1.34 +/- 0.21 vs. l.86 +/- 0.14 mm, relative light reflex amplitude 22.74 +/- 7.11% vs. 30.76 +/- 3.57%, v(constr) 3.75 +/- 1.09 vs. 5.80 +/- 0.59 mm/s) and of the parameters reflecting sympathetic pupillary function (diameter 5.69 +/- 0.66 vs. 6.35 +/- 0.60 mm, v(dil 1) 1.29 +/- 0.23 vs. 1.95 +/- 0.23 mm/s, v(dil 2) 0.64 +/- 0.13 vs. 0.72 +/ 0.l2 mm/s; Mann-Whitney U-test: p<0.05). The non-invasive technique of IPM demonstrates dysfunction not only of the cranial parasympathetic, but also of the cranial sympathetic nervous system and, thus, further characterizes autonomic dysfunction in FD. PMID- 11867079 TI - Relationship in the formation process between neurofibrillary tangles and Lewy bodies in the hippocampus of dementia with Lewy bodies brains. AB - Using tau immunohistochemistry and alpha-synuclein immunohistochemistry, we quantitatively investigated the most frequent sites and the formation process of neurofibrillary tangles (NFT) and Lewy bodies (LB) in the hippocampus from 20 patients with dementia with Lewy bodies (DLB). NFT were most frequently found in the CA2 and the subiculum-pre-CA1, while LB were most frequently found in the CA3 4 and the subiculum-pre-CA1. In the intrahippocampal routes of the perforant pathway, tau immunoelectron microscopy demonstrated distal axons containing aggregated tau-positive microtubules, while alpha-synuclein immunoelectron microscopy revealed terminal axons containing aggregated alpha-synuclein-positive tubular or filamentous components. These findings suggest that NFT and LB are first formed in the CA2 and the CA3-4 related to degeneration of the nonperforating route of the perforant pathway, respectively, and subsequently in the subiculum-pre-CA1 chiefly related to degeneration of the perforating route. Coexistence of NFT and LB in the same neurons was found most frequently in the subiculum-pre-CA1. In addition, coexistence of tau and alpha-synuclein was found in terminal axons of the perforant pathway, and tau accumulated not in paired helical filaments but in the periphery of alpha-synuclein-positive components immunoelectron-microscopically, suggesting that alpha-synuclein stimulates the accumulation of phosphorylated tau in terminal axons. PMID- 11867080 TI - Sharpening spots: correcting for bleedover in cDNA array images. AB - For cDNA array methods that depend on imaging of a radiolabel, we show that bleedover of one spot onto another, due to the gap between the array and the imaging media, can be a major problem. The images can be sharpened, however, using a blind convolution method based on the EM algorithm. The sharpened images look like a set of donuts, which concurs with our knowledge of the spotting process. Oversharpened images are actually useful as well, in locating the centers of each spot. PMID- 11867081 TI - Thresholding rules for recovering a sparse signal from microarray experiments. AB - We consider array experiments that compare expression levels of a high number of genes in two cell lines with few repetitions and with no subject effect. We develop a statistical model that illustrates under which assumptions thresholding is optimal in the analysis of such microarray data. The results of our model explain the success of the empirical rule of two-fold change. We illustrate a thresholding procedure that is adaptive to the noise level of the experiment, the amount of genes analyzed, and the amount of genes that truly change expression level. This procedure, in a world of perfect knowledge on noise distribution, would allow reconstruction of a sparse signal, minimizing the false discovery rate. Given the amount of information actually available, the thresholding rule described provides a reasonable estimator for the change in expression of any gene in two compared cell lines. PMID- 11867082 TI - A systematic statistical linear modeling approach to oligonucleotide array experiments. AB - We outline and describe steps for a statistically rigorous approach to analyzing probe-level Affymetrix GeneChip data. The approach employs classical linear mixed models and operates on a gene-by-gene basis. Forgoing any attempts at gene presence or absence calls, the method simultaneously considers the data across all chips in an experiment. Primary output includes precise estimates of fold change (some as low as 1.1), their statistical significance, and measures of array and probe variability. The method can accommodate complex experiments involving many kinds of treatments and can test for their effects at the probe level. Furthermore, mismatch probe data can be incorporated in different ways or ignored altogether. Data from an ionizing radiation experiment on human cell lines illustrate the key concepts. PMID- 11867083 TI - A simple method to improve probe set estimates from oligonucleotide arrays. AB - A popular commercially available oligonucleotide microarray technology employs sets of 25 base pair oligonucleotide probes for measurement of gene expression levels. A mathematical algorithm is required to compute an estimate of gene expression from the multiple probes. Previously proposed methods for summarizing gene expression data have either been substantially ad hoc or have relied on model assumptions that may be easily violated. Here we present a new algorithm for calculating gene expression from probe sets. Our approach is functionally related to leave-one-out cross-validation, a non-parametric statistical technique that is often applied in limited data situations. We illustrate this approach using data from our study seeking a molecular fingerprint of STAT3 regulated genes for early detection of human cancer. PMID- 11867084 TI - Multivariate approach for selecting sets of differentially expressed genes. AB - An important problem addressed using cDNA microarray data is the detection of genes differentially expressed in two tissues of interest. Currently used approaches ignore the multidimensional structure of the data. However it is well known that correlation among covariates can enhance the ability to detect less pronounced differences. We use the Mahalanobis distance between vectors of gene expressions as a criterion for simultaneously comparing a set of genes and develop an algorithm for maximizing it. To overcome the problem of instability of covariance matrices we propose a new method of combining data from small-scale random search experiments. We show that by utilizing the correlation structure the multivariate method, in addition to the genes found by the one-dimensional criteria, finds genes whose differential expression is not detectable marginally. PMID- 11867085 TI - Variable selection and pattern recognition with gene expression data generated by the microarray technology. AB - Lack of adequate statistical methods for the analysis of microarray data remains the most critical deterrent to uncovering the true potential of these promising techniques in basic and translational biological studies. The popular practice of drawing important biological conclusions from just one replicate (slide) should be discouraged. In this paper, we discuss some modern trends in statistical analysis of microarray data with a special focus on statistical classification (pattern recognition) and variable selection. In addressing these issues we consider the utility of some distances between random vectors and their nonparametric estimates obtained from gene expression data. Performance of the proposed distances is tested by computer simulations and analysis of gene expression data on two different types of human leukemia. In experimental settings, the error rate is estimated by cross-validation, while a control sample is generated in computer simulation experiments aimed at testing the proposed gene selection procedures and associated classification rules. PMID- 11867086 TI - Statistical inference for simultaneous clustering of gene expression data. AB - Current methods for analysis of gene expression data are mostly based on clustering and classification of either genes or samples. We offer support for the idea that more complex patterns can be identified in the data if genes and samples are considered simultaneously. We formalize the approach and propose a statistical framework for two-way clustering. A simultaneous clustering parameter is defined as a function theta=Phi(P) of the true data generating distribution P, and an estimate is obtained by applying this function to the empirical distribution P(n). We illustrate that a wide range of clustering procedures, including generalized hierarchical methods, can be defined as parameters which are compositions of individual mappings for clustering patients and genes. This framework allows one to assess classical properties of clustering methods, such as consistency, and to formally study statistical inference regarding the clustering parameter. We present results of simulations designed to assess the asymptotic validity of different bootstrap methods for estimating the distribution of Phi(P(n)). The method is illustrated on a publicly available data set. PMID- 11867087 TI - Dimension reduction strategies for analyzing global gene expression data with a response. AB - The analysis of global gene expression data from microarrays is breaking new ground in genetics research, while confronting modelers and statisticians with many critical issues. In this paper, we consider data sets in which a categorical or continuous response is recorded, along with gene expression, on a given number of experimental samples. Data of this type are usually employed to create a prediction mechanism for the response based on gene expression, and to identify a subset of relevant genes. This defines a regression setting characterized by a dramatic under-resolution with respect to the predictors (genes), whose number exceeds by orders of magnitude the number of available observations (samples). We present a dimension reduction strategy that, under appropriate assumptions, allows us to restrict attention to a few linear combinations of the original expression profiles, and thus to overcome under-resolution. These linear combinations can then be used to build and validate a regression model with standard techniques. Moreover, they can be used to rank original predictors, and ultimately to select a subset of them through comparison with a background 'chance scenario' based on a number of independent randomizations. We apply this strategy to publicly available data on leukemia classification. PMID- 11867088 TI - Temporal classification of Drosophila segmentation gene expression patterns by the multi-valued neural recognition method. AB - In order to reconstruct the establishment of the body pattern over time in Drosophila embryos, we have developed automated methods for detecting the age of an embryo on the basis of knowledge about its gene expression patterns. In this paper we perform temporal classification of confocal images of expression patterns of genes controlling segmentation by means of a neural network based on multi-valued neurons (MVN). MVN are artificial neural processing elements with complex-valued weights and high functionality, which proved to be efficient for solving the image recognition problems. The results obtained by this method confirm its efficiency for image recognition and indicate that the method can detect characteristic features of expression patterns which mark their development over time. PMID- 11867090 TI - The destruxins: synthesis, biosynthesis, biotransformation, and biological activity. AB - Destruxins, secondary metabolites first reported in 1961, are cyclic hexadepsipeptides composed of an alpha-hydroxy acid and five amino acid residues. The name "destruxin" is derived from "destructor" from the species Oospora destructor, the entomopathogenic fungus from which these metabolites were first isolated. Individual destruxins differ on the hydroxy acid, N-methylation, and R group of the amino acid residues; where established, the configurations of the amino acid residues are S, and those of the hydroxy acids are R. Destruxins exhibit a wide variety of biological activities, but are best known for their insecticidal and phytotoxic activities. The great interest in destruxins derives from their potential role as virulence factors in fungi, whether such microorganisms are useful insect biocontrol agents or detrimental, causing great plant disease epidemics. Reports on isolation, chemical structure determination, total synthesis, transformation by diverse organisms, and biological activity of destruxins and related metabolites are reviewed for the first time. PMID- 11867091 TI - De novo production of (+)-aristolochene by sporulated surface cultures of Penicillium roqueforti. AB - The de novo production of the fungal metabolite, (+)-aristolochene by sporulated surface cultures of Penicillium roqueforti is reported for the first time. The biosynthesis of fungal volatiles by various sporulated surface cultures was monitored by solid phase micro-extraction (SPME). When comparing malt extract agar with sabouraud dextrose agar, the highest yield of the fungal metabolite (0.04 mg/ml of culture) was obtained with the latter medium. The biosynthesis of (+)-aristolochene showed a maximum during the fourth day after inoculation. PMID- 11867092 TI - Nepenthes insignis uses a C2-portion of the carbon skeleton of L-alanine acquired via its carnivorous organs, to build up the allelochemical plumbagin. AB - Tropical pitcher plants (Nepenthes) catch animals in their specialized cup-shaped leaves, digest the prey by secreting enzymes, and actively take up the resulting compounds. The benefit of this behaviour is the ability to grow and compete in nutrient-poor habitats. Our present in vitro study shows that not only the nitrogen of alanine fed to the carnivorous organs is used by the plant but that in addition intact C2-units derived from C-2 and C-3 of stable isotope labelled L alanine serve as building blocks, here exemplarily for the synthesis of the secondary metabolite plumbagin, a potent allelochemical. This result adds a new facet to the benefit of carnivory for plants. The availability of plumbagin by a de novo synthesis probably enhances the plants' fitness in their defence against phytophagous and pathogenic organisms. A missing specific uptake or CoA activation mechanism might be the reason that acetate fed to the pitchers was not incorporated into the naphthoquinone plumbagin. The dihydronaphthoquinone glucosides rossoliside and plumbaside A, here isolated for the first time from Nepenthes, by contrast, showed no incorporation after feeding of any of the two precursors, suggesting these compounds to be storage forms with probably very low turnover rates. PMID- 11867093 TI - Chemical defenses of crucifers: elicitation and metabolism of phytoalexins and indole-3-acetonitrile in brown mustard and turnip. AB - The metabolism of the cruciferous phytoalexins brassinin and cyclobrassinin, and the related compounds indole-3-carboxaldehyde, glucobrassicin, and indole-3 acetaldoxime was investigated in various plant tissues of Brassica juncea and B. rapa. Metabolic studies with brassinin showed that stems of B. juncea metabolized radiolabeled brassinin to indole-3-acetic acid, via indole-3-carboxaldehyde, a detoxification pathway similar to that followed by the "blackleg" fungus (Phoma lingam/Leptosphaeria maculans). In addition, it was established that tetradeuterated brassinin was incorporated into the phytoalexin brassilexin in B. juncea and B. rapa. On the other hand, the tetradeuterated indole glucosinolate glucobrassicin was not incorporated into brassinin, although the chemical structures of brassinins and indole glucosinolates suggest an interconnected biogenesis. Importantly, tetradeuterated indole-3-acetaldoxime was an efficient precursor of phytoalexins brassinin, brassilexin, and spirobrassinin. Elicitation experiments in tissues of Brassica juncea and B. rapa showed that indole-3 acetonitrile was an inducible metabolite produced in leaves and stems of B. juncea but not in B. rapa. Indole-3-acetonitrile displayed antifungal activity similar to that of brassilexin, was metabolized by the blackleg fungus at slower rates than brassinin, cyclobrassinin, or brassilexin, and appeared to be involved in defense responses of B. juncea. PMID- 11867094 TI - Synthesis and bioactivity of C-2 and C-3 methyl ether derivatives of brassinolide. AB - The following six novel methyl ether derivatives of brassinolide were prepared and their brassinosteroid activity was measured by means of the rice leaf lamina inclination bioassay: 2-O-methylbrassinolide, 3-O-methylbrassinolide, 2,22,23-tri O-methylbrassinolide, 3,22,23-tri-O-methylbrassinolide, 2-O-methyl-25 methoxybrassinolide and 3-O-methyl-25-methoxybrassinolide. Brassinolide was used as a standard for comparison. All six compounds were also tested in the presence of 1000 ng of indole-3-acetic acid (IAA), an auxin that synergizes the effects of brassinosteroids. The 2-O-methyl- and 3-O-methylbrassinolide derivatives showed weak activity at high doses, which was enhanced by IAA, especially in the case of the 3-O-methyl derivative. Similarly, the 2,22,23-tri-O-methyl- and 3,22,23-tri-O methyl derivatives displayed weak bioactivity on their own, but significantly stronger activity when applied with IAA. The 3-O-methyl and 3,22,23-tri-O-methyl analogues plus IAA were comparable in bioacivity to brassinolide alone, but were less active than brassinolide plus IAA. In each case, O-methylation at C-2 resulted in a greater loss of activity than O-methylation at C-3 under the same conditions. The relatively strong activity of 3,22,23-tri-O-methylbrassinolide in the presence of IAA is especially noteworthy as it indicates that free hydroxyl groups at C-3, C-22, and C-23 are not essential for bioactivity. Finally, 2-O methyl- and 3-O-methyl-25-methoxybrassinolide were essentially inactive alone, and showed only a modest increase in bioactivity when coapplied with IAA. PMID- 11867096 TI - Gerronemins A-F, cytotoxic biscatechols from a Gerronema species. AB - The gerronemins A-F (1-6) were isolated as the cytotoxic components of an extract of a Gerronema species detected in a screening for new cytotoxic metabolites from basidiomycetes. Their structures were elucidated by spectroscopic techniques, and they are composed of a C12-C16 alkane or alkene substituted at both ends by 2,3 dihydroxyphenyl groups. The gerronemins blocked the inducible expression of a hCOX-2 and iNOS promoter driven reporter gene with IC50-values of 1-5 microg/ml. In addition, cytotoxic activities were observed which were due the inhibition of cellular macromolecular syntheses. PMID- 11867095 TI - Cytotoxic withanolides from Acnistus arborescens. AB - Three cytotoxic withanolides, two with new structures, were isolated from the leaves of Acnistus arborescens and their structures determined by a combination of 1D and 2D NMR, mass spectral, and molecular modeling studies. Dereplication analysis of the ethyl ether extract was useful for evaluating the components showing significant cytotoxic activity. PMID- 11867097 TI - Coumarins and gamma-pyrone derivatives from Prangos pabularia: antibacterial activity and inhibition of cytokine release. AB - The n-hexane and ethyl acetate extracts of the stems and the ethyl acetate extracts of roots from Prangos pabularia afforded an gamma-pyrone derivative and furanocoumarin derivatives with three glucose and gamma-pyrone (pabularin A, B and C), along with 26 previously known compounds (18 coumarins, six terpenoids and two glycosides). Their structures were established on the basis of spectroscopic studies. Of these, 16 coumarin derivatives isolated from P. pabularia were tested for antibacterial activity and inhibition of cytokine release. PMID- 11867099 TI - Benzoic acid glucosinolate esters and other glucosinolates from Arabidopsis thaliana. AB - The spectacular recent progress in Arabidopsis thaliana molecular genetics furnishes outstanding tools for studying the formation and function of all metabolites in this cruciferous species. One of the major groups of secondary metabolites in A. thaliana is the glucosinolates. These hydrophilic, sulfur-rich glycosides appear to serve as defenses against some generalist herbivores and pathogens, and as feeding and oviposition stimulants to specialist herbivores. To help study their biosynthesis and role in plant-insect interactions, we wanted to determine the complete glucosinolate content of A. thaliana. In previous studies, 24 glucosinolates had been identified from ecotype Columbia. We reinvestigated Columbia as well as additional ecotypes and mutant lines, and identified 12 further glucosinolates, including five novel compounds. Structures were elucidated by MS and NMR spectroscopy of their desulfated derivatives, and by enzymatic cleavage of the attached ester moieties. Four of the novel glucosinolates are benzoate esters isolated from the seeds. In all but one of these compounds, esterification is on the glucose moiety rather than the side chain, a very unusual feature for glucosinolates. Among additional glucosinolates identified were the first non-chain elongated, methionine-derived glucosinolate from A. thaliana and the first compounds that appear to be derived from leucine. PMID- 11867098 TI - Volatiles from rhizomes of Rhodiola rosea L. AB - Terpenes and aroma volatiles from rhizomes of Rhodiola rosea L. from Norway have been isolated by both steam distillation and headspace solid-phase micro extraction coupled with gas chromatography and mass spectrometry analysis. The dried rhizomes contained 0.05% essential oil with the main chemical classes: monoterpene hydrocarbons (25.40%), monoterpene alcohols (23.61%) and straight chain aliphatic alcohols (37.54%). n-Decanol (30.38%), geraniol (12.49%) and 1,4 p-menthadien-7-ol (5.10%) were the most abundant volatiles detected in the essential oil, and a total of 86 compounds were identified in both the SD and HS SPME samples. Geraniol was identified as the most important rose-like odour compound besides geranyl formate, geranyl acetate, benzyl alcohol and phenylethyl alcohol. Floral notes such as linalool and its oxides, nonanal, decanal, nerol and cinnamyl alcohol highlight the flowery scent of rose root rhizomes. PMID- 11867101 TI - Gibberellins in shoots and developing capsules of Populus species. AB - Extracts of stems of growing shoots of Populus deltoides and P. trichocarpa, and developing capsules of P. deltoides were analysed for gibberellins (GAs) by gas chromatography-mass spectrometry. The following known GAs were identified by comparison of their Kovats retention indices (KRIs) and mass spectra with those of standards: GA1, GA8, GA9, GA19, GA20, 16 beta,17-dihydro-17-hydroxy GA20, GA23, GA28, GA29, GA34, GA44, and GA97. Several of these have not been previously reported from Populus. In addition, two new GAs were identified as 12 beta hydroxy GA53 (GA127) and 16 beta,17-dihydro-17-hydroxy GA53 and their structures were confirmed by partial synthesis. Evidence was found of 16,17-dihydro-16,17 dihydroxy GA9, 16,17-dihydro-16,17-dihydroxy GA12, 12-hydroxy GA14, and GA34 catabolite by comparison of mass spectra and KRIs with published data. Several putative GAs (hydroxy- and dihydroxy-GA12-like) were also found. The catabolites of active GAs or of key precursors, hydroxylated at C-2 in stems and either C-2, C-12, C-17, or C-16,17 in capsules, were the major proportion of the GAs. PMID- 11867100 TI - Structure and stereochemistry of dimeric proteracacinidins possessing the rare C 4(C) --> C-5(D) interflavanyl linkage. AB - The rare series of (4-->5)-linked proteracacinidins is extended by identification of oritin-(4alpha-->5)-epioritin-4beta-ol, ent-epioritin-(4alpha-->5)-epioritin 4beta-ol, epioritin-(4beta-->5)-epioritin-4alpha-ol and ent-oritin-(4beta-->5) epioritin-4alpha-ol from the heartwoods of Acacia galpinii and Acacia caffra. PMID- 11867102 TI - Undermining international family-planning programmes. PMID- 11867104 TI - Linear growth retardation and cognition. PMID- 11867103 TI - Adverse reactions to TNF-alpha inhibitors in rheumatoid arthritis. PMID- 11867105 TI - Role of intracoronary brachytherapy for in-stent restenosis? PMID- 11867106 TI - Serious mental disorder in 23000 prisoners: a systematic review of 62 surveys. AB - BACKGROUND: About 9 million people are imprisoned worldwide, but the number with serious mental disorders (psychosis, major depression, and antisocial personality disorder) is unknown. We did a systematic review of surveys on such disorders in general prison populations in western countries. METHODS: We searched for psychiatric surveys that were based on interviews of unselected prison populations and included diagnoses of psychotic illnesses or major depression within the previous 6 months, or a history of any personality disorder. We did computer-assisted searches, scanned reference lists, searched journals, and corresponded with authors. We determined prevalence rates of serious mental disorders, sex, type of prisoner (detainee or sentenced inmate), and other characteristics. FINDINGS: 62 surveys from 12 countries included 22790 prisoners (mean age 29 years, 18530 [81%] men, 2568 [26%] of 9776 were violent offenders). 3.7% of men (95% CI 3.3--4.1) had psychotic illnesses, 10% (9--11) major depression, and 65% (61--68) a personality disorder, including 47% (46--48) with antisocial personality disorder. 4.0% of women (3.2--5.1) had psychotic illnesses, 12% (11--14) major depression, and 42% (38--45) a personality disorder, including 21% (19--23) with antisocial personality disorder. Although there was substantial heterogeneity among studies (especially for antisocial personality disorder), only a small proportion was explained by differences in prevalence rates between detainees and sentenced inmates. Prisoners were several times more likely to have psychosis and major depression, and about ten times more likely to have antisocial personality disorder, than the general population. INTERPRETATION: Worldwide, several million prisoners probably have serious mental disorders, but how well prison services are addressing these problems is not known. PMID- 11867107 TI - Use of localised intracoronary beta radiation in treatment of in-stent restenosis: the INHIBIT randomised controlled trial. AB - BACKGROUND: In-stent restenosis is a major limitation of intracoronary stenting. Ionising gamma radiation has been shown to reduce recurrence of restenosis after stent placement. We aimed to compare the effects of intracoronary beta radiation treatment with those of placebo for clinical and angiographic outcomes of patients with diffuse in-stent restenosis. METHODS: 332 patients with in-stent restenosis underwent successful coronary intervention, and were then randomly allocated to intracoronary beta radiation with a phosphorus-32 source (n=166) or placebo (166) delivered into a centreing balloon catheter through an automatic afterloader. Longer lesions (>22 mm of dilated length) were treated with tandem positioning of the study wire. The primary safety endpoint was major adverse cardiac events, defined as death, myocardial infarction, and repeat target-lesion revascularisation at 9 months. The primary efficacy endpoint was binary angiographic restenosis rate in the analysis segment during 9-months' follow-up. Analysis was by intention to treat. FINDINGS: Procedural success, and in-hospital and 30-day complications were similar among the two groups. 24 (15%) patients in the radiated group had the primary safety endpoint of death, myocardial infarction, or repeat target-lesion revascularisation over 290 days compared with 51 [corrected] (31%) in the placebo group (difference 16% [95% CI 7-25], p = 0.0006). Binary angiographic restenosis rate was lower in the radiated group than the placebo group for the entire analysed segment (difference 25% [14--37], p < 0.0001). INTERPRETATION: Vascular brachytherapy using pure beta-emitter 32P delivered into a centreing catheter via an automatic afterloader can be used to reduce overall revascularisation in patients undergoing treatment for diffuse in stent restenosis. PMID- 11867108 TI - Clinical picture: Takayasu's arteritis. PMID- 11867109 TI - Blood lactate as an early predictor of outcome in paracetamol-induced acute liver failure: a cohort study. AB - BACKGROUND: Although the King's College Hospital (KCH) selection criteria for emergency liver transplantation in paracetamol-induced acute liver failure are widely used, strategies to improve sensitivity and facilitate earlier transplantation are required. We investigated the use of arterial blood lactate measurement for the identification of transplantation candidates. METHODS: In a single-centre study, we measured arterial blood lactate early (median 4 h) and after fluid resuscitation (median 12 h) in patients admitted to a tertiary referral intensive-care unit. Threshold values that best identified individuals likely to die without transplantation were derived in a retrospective initial sample of 103 patients with paracetamol-induced acute liver failure and applied to a prospective validation sample of 107 patients. Predictive value and speed of identification were compared with those of KCH criteria. FINDINGS: In the initial sample, median lactate was significantly higher in non-surviving patients than in survivors both in the early samples (8.5 [range 1.7--21.0] vs 1.4 [0.53--7.9] mmol/L, p<0.0001) and after fluid resuscitation (5.5 [1.3--18.6] vs 1.3 [0.26- 3.2], p<0.0001). Applied to the validation sample, a threshold value of 3.5 mmol/L early after admission had sensitivity 67%, specificity 95%, positive likelihood ratio 13, and negative likelihood ratio 0.35; the corresponding values for a threshold of 3.0 mmol/L after fluid resuscitation were 76%, 97%, 30, and 0.24. Combined early and postresuscitation lactate concentrations had similar predictive ability to KCH criteria but identified non-surviving patients earlier (4 [3--13] vs 10 [3.5--19.5] h, p=0.01). Addition of postresuscitation lactate concentration to KCH criteria increased sensitivity from 76% to 91% and lowered negative likelihood ratio from 0.25 to 0.10. INTERPRETATION: Arterial blood lactate measurement rapidly and accurately identifies patients who will die from paracetamol-induced acute liver failure. Its use could improve the speed and accuracy of selection of appropriate candidates for transplantation. PMID- 11867110 TI - Effects of stunting, diarrhoeal disease, and parasitic infection during infancy on cognition in late childhood: a follow-up study. AB - BACKGROUND: Chronic malnutrition during infancy, marked by stunting, has been associated with poor cognitive function. We assessed the effect of stunting, diarrhoeal disease, and parasitic infections during infancy on cognitive function in late childhood. METHODS: We followed up from birth to 2 years, a cohort of 239 Peruvian children for anthropometrics, stool samples, and diarrhoeal status. At 9 years of age, we assessed cognitive function in 143 (69%) with the full-scale intelligence quotient of the Wechsler intelligence scale for children-revised (WISC-R). Findings All findings were adjusted for socioeconomic status and schooling; in addition, findings related to diarrhoea prevalence, Giardia lamblia, and Cryptosporidium parvum were adjusted for severe stunting. During the first 2 years of life, 46 (32%) of 143 children were stunted. Children with severe stunting in the second year of life scored 10 points lower on the WISC-R test (95% CI 2.4--17.5) than children without severe stunting. Children with more than one episode of G lamblia per year scored 4.1 points (0.2--8.0) lower than children with one episode or fewer per year. Neither diarrhoea prevalence nor Cparvum infection was associated with WISC-R scores. INTERPRETATION: Malnutrition in early childhood, indexed by stunting, and potentially G lamblia, are associated with poor cognitive function at age 9 years. If the observed associations are causal, then intervention programmes designed to prevent malnutrition and G lamblia early in life could lead to significant improvement in cognitive function of children in similar lower-income communities throughout the less-developed world. PMID- 11867112 TI - Use of proteomic patterns in serum to identify ovarian cancer. AB - BACKGROUND: New technologies for the detection of early-stage ovarian cancer are urgently needed. Pathological changes within an organ might be reflected in proteomic patterns in serum. We developed a bioinformatics tool and used it to identify proteomic patterns in serum that distinguish neoplastic from non neoplastic disease within the ovary. METHODS: Proteomic spectra were generated by mass spectroscopy (surface-enhanced laser desorption and ionisation). A preliminary "training" set of spectra derived from analysis of serum from 50 unaffected women and 50 patients with ovarian cancer were analysed by an iterative searching algorithm that identified a proteomic pattern that completely discriminated cancer from non-cancer. The discovered pattern was then used to classify an independent set of 116 masked serum samples: 50 from women with ovarian cancer, and 66 from unaffected women or those with non-malignant disorders. FINDINGS: The algorithm identified a cluster pattern that, in the training set, completely segregated cancer from non-cancer. The discriminatory pattern correctly identified all 50 ovarian cancer cases in the masked set, including all 18 stage I cases. Of the 66 cases of non-malignant disease, 63 were recognised as not cancer. This result yielded a sensitivity of 100% (95% CI 93- 100), specificity of 95% (87--99), and positive predictive value of 94% (84--99). INTERPRETATION: These findings justify a prospective population-based assessment of proteomic pattern technology as a screening tool for all stages of ovarian cancer in high-risk and general populations. PMID- 11867113 TI - A seropneumothorax? PMID- 11867114 TI - Drug-induced systemic lupus erythematosus associated with etanercept therapy. AB - Specific antagonists of tumour necrosis factor (TNF)-alpha have rapidly gained popularity for the treatment of rheumatoid arthritis. The monoclonal antibody against TNF-alpha, infliximab, has been associated with induction of systemic lupus erythematosus (SLE); however, there have been no published reports of drug induced SLE associated with the soluble TNF-alpha receptor etanercept. We describe four female patients who developed signs and symptoms of SLE during treatment with etanercept; in two SLE was unambiguous. On diagnosis of SLE, etanercept was discontinued and the SLE-related symptoms promptly resolved. Etanercept should be considered in the list of agents associated with drug induced SLE. PMID- 11867115 TI - Early growth, menarche, and adiposity rebound. AB - Adiposity rebound is thought to arise earlier in children with a high body-mass index (BMI) at age 3 years. To test this hypothesis we followed up a birth cohort. Our results indicate that earlier rebound arose in children who were tall at age 3 years, but that timing was not associated with BMI at age 3 years in boys, and arose later in girls with higher BMIs at age 3. Adiposity rebound also arose earlier in children whose parents had high rather than low BMIs. Magnitude of BMI at rebound was associated with height and BMI at age 3, and with parental BMI. There was a correlation between age at rebound and age at menarche, which suggests that timing of rebound is an indicator of physical maturity rather than obesity. PMID- 11867116 TI - W135 meningococcal disease in England and Wales associated with Hajj 2000 and 2001. AB - An outbreak of W135 meningococcal disease was seen in 2000 and 2001 among pilgrims returning from Saudi Arabia and their contacts. The Public Health Laboratory Service set up enhanced surveillance to monitor spread and virulence of the outbreak strain, and to collect data on case characteristics. The number of cases reported from England and Wales in 2001 was similar to the number reported during 2000, despite a change in the recommendation for pilgrims to receive quadrivalent vaccine (against serogroup A, C, W135, and Y) instead of the A/C vaccine recommended in 2000. Both the recommendation and vaccine were not available until January, 2001, and coverage among pilgrims to the Hajj 2001 was low. The case--fatality ratio was high, and sustained transmission of the virulent outbreak strain was seen. Current control policies, both in primary and secondary prevention, might need better implementation. PMID- 11867118 TI - European tobacco control reaches a critical phase. PMID- 11867119 TI - Cloned mice have a shorter lifespan than "normal" mice. PMID- 11867117 TI - Risk of early febrile seizure with perinatal exposure to nucleoside analogues. AB - The frequency of seizures was studied in a prospective cohort of French children born to HIV-1-infected mothers. The analysis was restricted to the 4426 uninfected children, whether or not exposed to antiretrovirals. 81 convulsions were reported up to the age of 18 months, and 30 children fulfilled the criteria for simple febrile seizures. The risk of first febrile seizure was higher for children perinatally exposed to antiretrovirals than for those not exposed (log rank test: p=0.0198). A similar trend was noted for other non-neonatal seizures (p=0.0537) but not for neonatal seizures. PMID- 11867120 TI - Researchers identify a possible new target for treatment of malaria. PMID- 11867121 TI - Health checks before facing tourism's final frontier. PMID- 11867122 TI - A season of U-turns for UK health politics. PMID- 11867123 TI - Dutch investigators recommend prescription of heroin to addicts. PMID- 11867126 TI - What role should the EU have in health-care regulation? PMID- 11867127 TI - Health ministers support pan-European transplantation standards. PMID- 11867128 TI - South African President hints at expansion of HIV-drug pilot programmes. PMID- 11867129 TI - Experts call for surveillance of drug-resistant typhoid at a global level. PMID- 11867130 TI - Atrial fibrillation: strategies to control, combat, and cure. AB - Atrial fibrillation is the commonest clinical arrhythmia, is increasing in incidence and prevalence, and is associated with substantial morbidity and mortality. The arrhythmia may be paroxysmal (self-limiting), persistent (amenable to cardioversion), or permanent. Especially in its paroxysmal form, atrial fibrillation may be initiated by rapidly firing foci, generally located in the proximal pulmonary veins. Sustained atrial fibrillation is maintained by an atrial tissue substrate capable of accommodating many meandering wavelets. With continuing arrhythmia, the electrophysiological properties of the atria change and further facilitate continuing fibrillation. Treatment is aimed at prevention of thromboembolic complications, restoration and maintenance of sinus rhythm, and control of ventricular rate during atrial fibrillation. With greater understanding of the arrhythmia mechanisms, it is becoming possible to offer targeted curative treatments to more and more patients. PMID- 11867131 TI - Liver and kidney preservation by perfusion. AB - The clinical boundaries of transplantation have been set in an era of simple cold storage. Research in organ preservation has led to the development of flush solutions that buffer the harsh molecular conditions which develop during ischaemia, and provide stored organs that are fit to sustain life after transplantation. Although simple and efficient, this method might be reaching its limit with respect to the duration, preservation, and the quality of organs that can be preserved. In addition, flush preservation does not allow for adequate viability assessment. There is good evidence that preservation times will be extended by the provision of continuous cellular substrate. Stimulation of in vivo conditions by ex-vivo perfusion could also mean that marginal organs will be salvaged for transplantation. Perfusion will also allow for assessing the viability of organs before transplantation in a continuous fashion. The cumulative effect of these benefits would include expansion of the donor pool, less risk of primary non-function, and extension of the safe preservation period. Use of non-heart-beating donors, international organ sharing, and precise calculation of the risk of primary organ failure could become standard. PMID- 11867132 TI - Allocation concealment in randomised trials: defending against deciphering. AB - Proper randomisation rests on adequate allocation concealment. An allocation concealment process keeps clinicians and participants unaware of upcoming assignments. Without it, even properly developed random allocation sequences can be subverted. Within this concealment process, the crucial unbiased nature of randomised controlled trials collides with their most vexing implementation problems. Proper allocation concealment frequently frustrates clinical inclinations, which annoys those who do the trials. Randomised controlled trials are anathema to clinicians. Many involved with trials will be tempted to decipher assignments, which subverts randomisation. For some implementing a trial, deciphering the allocation scheme might frequently become too great an intellectual challenge to resist. Whether their motives indicate innocent or pernicious intents, such tampering undermines the validity of a trial. Indeed, inadequate allocation concealment leads to exaggerated estimates of treatment effect, on average, but with scope for bias in either direction. Trial investigators will be crafty in any potential efforts to decipher the allocation sequence, so trial designers must be just as clever in their design efforts to prevent deciphering. Investigators must effectively immunise trials against selection and confounding biases with proper allocation concealment. Furthermore, investigators should report baseline comparisons on important prognostic variables. Hypothesis tests of baseline characteristics, however, are superfluous and could be harmful if they lead investigators to suppress reporting any baseline imbalances. PMID- 11867133 TI - Overcoming restenosis with sirolimus: from alphabet soup to clinical reality. PMID- 11867134 TI - Asthma and farming. PMID- 11867136 TI - Community-based approach to HIV treatment. PMID- 11867137 TI - Pathological correlates of dementia. PMID- 11867138 TI - Early Pseudomonas aeruginosa colonisation in cystic fibrosis patients. PMID- 11867141 TI - Early Pseudomonas aeruginosa colonisation in cystic fibrosis patients. PMID- 11867142 TI - Neonatal screening for medium-chain acyl-CoA dehydrogenase deficiency. PMID- 11867143 TI - Chagas' disease. PMID- 11867144 TI - Neonatal screening for medium--chain acyl-CoA dehydrogenase deficiency. PMID- 11867145 TI - Hyperkalaemia in a patient with diabetes mellitus. PMID- 11867146 TI - Norwegian obstetricians prefer vaginal route of delivery. PMID- 11867147 TI - Encouragement of cancer patients. PMID- 11867148 TI - The future of nuclear medicine. PMID- 11867150 TI - New law on male circumcision in Sweden. PMID- 11867151 TI - Careful naming of disorders. PMID- 11867161 TI - A medical education. PMID- 11867162 TI - The highest priority: what microbial genomes are telling us about immunity. AB - Study of microbial genomes has provided new insight into the functions that pathogens require for survival in the animal host. Small genome bacterial pathogens, defined as those < or = 1/3 the size of Escherichia coli, include chlamydiae, rickettsiae and ehrlichiae, mycoplasmas, and spirochetes. The small genome size is believed to result from reductive evolution, a process of initial mutation with loss of function followed by progressive accumulation of mutations and eventual gene deletion. This is most notable in the 1.1 Mb genome of Rickettsia prowazekki in which 24% of the genome is non-coding, as compared to approximately 10% in the 4.4 Mb E. coli. Consequently, these pathogens are thus presumed to retain only the most important functions for survival and propagation. There is consistent evidence from small genomes that the genetic deletion is primarily related to the loss of metabolic function and especially reduction of multiple overlapping pathways and duplicated genes. Thus, these pathogens undergo progressive reduction in their genomes yet maintain the ability to infect, survive within, and cause disease in animals. In the face of this reductive process, what genes and associated functions are maintained? Strikingly, these pathogens devote a high percentage of their genomes to paralogous families of polymorphic surface molecules. This retention suggests that evasion of the immune response is the highest priority of obligate microbial pathogens and provides a strategy for identifying protective antigens for vaccine development to control disease. PMID- 11867163 TI - Characterization of a feline homologue of the alphaE integrin subunit (CD103) reveals high specificity for intra-epithelial lymphocytes. AB - The characteristics of a feline homologue of the alphaE integrin (CD103), defined by two murine monoclonal antibodies, Fe7.1B8 (IgG1) and Fe7.2D8 (IgG1), are described. These antibodies recognized 75% of intra-epithelial (range 59-88%) and 40% of lamina proprial (range 28-46%) T cells of the intestinal mucosal tissue of the small intestine in contrast with approximately 2% of peripheral blood lymphocytes. Both antibodies immunoprecipitated a 180 kDa protein from biotinylated feline intra-epithelial mucosal leukocytes consistent with the alphaE integrin subunit in conjunction with a 120 kDa protein consistent with the beta7 subunit. The nucleotide sequence of feline alphaE integrin, generated from molecular cloning of the feline alphaE encoding cDNA, is also reported. This feline molecule shares 72% sequence homology with human and 69% homology with murine and rat counterparts. Homology includes the presence of an X (extra) domain, that appears unique to alphaE molecules as described for human, rat and mouse, as well as areas of homology common to other alpha integrins. Of note is a typical I (inserted) domain, the presence of seven repeat regions, and highly conserved sequences in the cytoplasmic tail. Transfection studies demonstrated that both antibodies recognized an extracellular component which encompassed the X and I domains of the cloned alphaE integrin subunit. These studies demonstrate that the pattern of tissue distribution, biochemical characteristics, and cDNA sequence of the feline alphaE integrin subunit are largely similar to that described for other species. PMID- 11867165 TI - Uptake of colostral leukocytes in the intestinal tract of newborn calves. AB - The role of colostral immunoglobulins for the protection of newborn calves has been studied extensively, but little is known about the importance of colostral leukocytes. To study the uptake of colostral leukocytes in the intestine of calves and to determine preferential sites for this uptake, FITC-labelled colostral cells derived from the respective dams were injected into intestinal loops with/without Peyer's patches of three male Holstein Frisian calves about 5h post natum. In adjacent loops, PBS was injected as control. Loops were excised after an exposure of 1.5-2h. FITC-labelled material and cells were detected by the direct immunoperoxidase method in paraplast sections. Twenty-five consecutive sections were evaluated from each localization. Uptake of labelled material and cells was observed in all three calves in the jejunal Peyer's patch and in two calves in the ileal Peyer's patch as well. In the jejunal Peyer's patch, labelled material and cells were present in epithelium, domes and sinuses around lymphoid follicles, whereas in the ileal Peyer's patch, they were found in the sinuses only. These findings confirm that uptake of colostral leukocytes through the intestinal barrier is possible and that the preferential route of uptake is through follicle-associated epithelium of Peyer's patches. PMID- 11867164 TI - Immunisation of cattle against heartwater by infection with Cowdria ruminantium elicits T lymphocytes that recognise major antigenic proteins 1 and 2 of the agent. AB - There is growing evidence that immunity of cattle to Cowdria ruminantium infection is mediated by T lymphocytes. C. ruminantium antigens that stimulate these responses are therefore of considerable importance to the development of a sub-unit vaccine against the disease. We have examined T cell responses against recombinant analogues of the surface-exposed C. ruminantium major antigen 1 (MAP1) a 28.8 kDa protein and MAP2 (21 kDa) antigen in cattle immunised by infection and treatment. Vigorous and sustained proliferative responses to both antigens were observed in peripheral blood mononuclear cells from immune cattle. MAP1-specific responses were predominantly restricted to cluster of differentiation four antigen positive T cells (CD4+ T cells). Reverse transcription polymerase chain reaction (RT-PCR) analysis of cytokine expression by T cell lines derived from this population revealed strong expression of interferon gamma (IFN-gamma), interferon alpha (IFN-alpha), tumour necrosis factor alpha (TNF-alpha), tumour necrosis factor beta (TNF-beta), interleukin-2 receptor alpha (IL-2Ralpha) transcripts, and weak expression of IL-2 and IL-4. Supernatants from these T cell cultures contained IFN-gamma protein. CD4+ T cell clones specific for MAP1 were generated. Two of these clones proliferated in the presence of autologous infected endothelial cells. In contrast, the response to MAP2 was characterised largely by proliferation of gamma delta (gammadelta) T cells. RT-PCR analysis of cytokine expression by T cell lines which were dominated by gammadelta T cells revealed expression of IFN-gamma, TNF-alpha, TNF beta, IL-2Ralpha transcripts. Supernatants of these T cell cultures also contained IFN-gamma protein. Our findings indicate that immunisation of cattle by infection with C. ruminantium results in generation of MAP1- and MAP2-specific T cell responses that may play a role in protection against the pathogen. PMID- 11867166 TI - Oral administration of yeast, Saccharomyces cerevisiae, enhances the cellular innate immune response of gilthead seabream (Sparus aurata L.). AB - The effects of including lyophilised whole yeast, Saccharomyces cerevisiae, in the diet on the seabream innate immune response were investigated. Gilthead seabream (Sparus aurata L.) specimens were fed four different diets for 4 weeks: a commercial diet as control and the same diet supplemented with 1, 5 or 10 g/kg yeast. After 1, 2 and 4 weeks, serum complement titres, as a humoral parameter, and phagocytic, respiratory burst, myeloperoxidase and natural cytotoxic activities of head-kidney leucocytes, as cellular parameters, were evaluated. The results showed that yeast supplements enhanced all the latter responses, but not the humoral response. This enhancement was dose-dependent except for the cytotoxic activity that was only stimulated by the lower dose of yeast assayed. As yeast cell walls are able to enhance the seabream cellular innate immune response, these results support the possible use of whole yeast as natural inmunostimulants in common fish diets. PMID- 11867167 TI - Cellular immune response in the small intestine of two broiler chicken lines orally inoculated with malabsorption syndrome homogenates. AB - We studied the cellular immune response against malabsorption syndrome (MAS) in two broiler chicken lines, A and B. We determined the number of pan T-lymphocytes (CD3), helper T-lymphocytes (CD4), cytotoxic T-lymphocytes (CD8) and macrophages/monocytes in the small intestine in the first 2 weeks after oral inoculation of two MAS homogenates, MAS80 and MAS97-1. The immune cells were detected on cryostat tissue by immunohistochemistry and counted by villus area. In trial 1, we compared the two broiler lines for weight gain depression, intestinal lesion and number of CD3, CD4, CD8 cells and macrophages/monocytes after MAS80 inoculation. Although there was no significant difference in weight gain depression between the two broiler lines, line B had significantly higher numbers of CD8+ T-cells per villus area than had line A. To confirm part of the results of trial 1, trial 2 was done in which we compared different homogenates in broiler line B. Broiler line B was orally inoculated with either MAS97-1, intestinal homogenate obtained from healthy chickens (healthy homogenate), or phosphate buffered saline (PBS). In this trial, the MAS97-1 homogenate also induced weight gain depression and intestinal lesions, whereas the "healthy homogenate" and PBS did not induce weight gain depression or intestinal lesions. The broilers inoculated with MAS97-1 homogenate had significantly more CD8+ T cells per villus area than had broilers inoculated with "healthy homogenate" or PBS. Increased CD8+ T-cells per villus area in the affected small intestines of broilers suggests an increase of cytotoxic T-cell activity. PMID- 11867168 TI - Similar pattern of iNOS expression, NO production and cytokine response in genetic and vaccination-acquired resistance to Marek's disease. AB - NO is produced by macrophages through activation of the inducible enzyme NOS and its production is triggered as an antiviral and antitumoral immune mechanism. Replication of Marek's disease herpes virus (MDV) is inhibited by NO in vitro. MDV induces T-lymphomas in the chicken and a genetic resistance to tumor development has been linked to the B21 major histocompatibility complex. During the first initial week of viral replication after inoculation of the highly virulent RB-1B MDV strain, histocompatible B21/B21 chickens developed strong iNOS expression and NO production capacity in the spleen, in parallel with strong systemic NO production in the serum. Comparable NO response was not seen with the vaccinal strain HVT. In contrast, reduction in spleen macrophage number and delay in iNOS gene expression was observed in genetically susceptible B13/B13 chickens after MDV infection, in addition to suppression of IFN-gamma-inducible NO production. However, vaccination with HVT 3 days before RB-1B inoculation restored strong iNOS gene expression in the spleen 1 week later and inducible NO production 3 weeks later. Following the pattern of iNOS gene expression, early strong expression of cytokines with powerful iNOS-inducing activity such as IFN gamma and CC chemokines from the MIP family (MIP-1beta, K203) was observed in genetic resistance and resistance acquired after vaccination with HVT. In conclusion, resistance to MDV appeared preferentially linked in both types of resistance to the early establishment of cytokine induction characteristic of a Th1 immune response, thus favoring the development of an early and strong NO response. PMID- 11867169 TI - Differential alloreactivity at SLA-DR and -DQ matching in two-way mixed lymphocyte culture. AB - The major histocompatibility complex (MHC) class II molecules are heterodimeric cell surface glycoproteins important for antigen presentation to CD4+ T lymphocytes. Class II molecules of the pig MHC, termed SLA, identified so far include DR and DQ. Thus far, functional differences between products of different loci in SLalpha class II have not been well characterized. For detailed research on this issue, SLalpha-DRbeta1 and -DQbeta typings were newly developed by restriction fragment length polymorphism (RFLP) of the reverse transcriptase polymerase chain reaction (RT-PCR) products. Using this method, several RFLP types were chosen from 13 CSK miniature pigs, and alloreactivities in two-way mixed lymphocyte culture (MLC) derived from these pigs were examined by cell proliferation assay using flow cytometry. The responses in MLC varied according to the degree of phenotype difference. In MLC from individuals of the same RFLP type in both SLA-DRbeta1 and -DQbeta, the proliferative responses showed slight reaction indicating that they were not so stimulated by each other. On the other hand, for the RFLP type-mismatching combination, the responses were strong indicating that they recognized each others alloantigens. The reactivity of only the DQbeta mismatching combination was as strong as those of only the DRbeta1 mismatching combination. These data indicate the important role of the DQ as well as DR molecule on alloreactivity in MLC. PMID- 11867170 TI - Mucosal and systemic isotype-specific antibody responses and protection in conventional pigs exposed to virulent or attenuated porcine epidemic diarrhoea virus. AB - Eleven-day-old conventionally reared piglets were inoculated orally with two different doses of the cell-culture adapted strain CV-777 of the porcine epidemic diarrhoea virus (PEDV) or the virulent isolate of the same strain and challenged with the same virulent PEDV 3 weeks later. Pigs inoculated with the two doses of the attenuated virus did not show any typical sign of the disease, and virus shedding was not frequent. In contrast, 31% of pigs exposed to the virulent PEDV developed diarrhoea and virus shedding was demonstrated in 100%. At different postinoculation day (PID) and postchallenge day (PCD) virus-specific antibody secreting cells (ASC) in gut associated lymphoid tissues (duodenum and ileum lamina propria and mesenteric lymph nodes) and systemic locations (blood and spleen) were assessed by enzyme-linked immunospot (ELISPOT). Only a small response was detected in the groups inoculated with attenuated PEDV, whereas in the group previously exposed to the virulent virus on PID 21 a large number of IgG and IgA ASC was detected. Isotype-specific antibody responses in serum were investigated by ELISA. IgG responses were detected in all groups, although the highest response corresponded to the group inoculated with virulent virus and only this group showed an IgA response. The pigs exposed to virulent PEDV were completely protected against the challenge with a higher dose of the same virulent virus on PID 21 and none of them shed the virus. The pigs inoculated with the attenuated strain were partially protected against the challenge, and 25% of the low dose- and 50% of the high dose-exposed pigs did not shed virus after challenge. All the pigs from a control group, not previously exposed to the virus, excreted the virus in faeces. A strong positive correlation was established between protection and the ASC responses detected in gut associated lymphoid tissues and blood at the challenge day and also between protection and serum isotype-specific antibody titers on that day. In addition, the IgA and IgG ASC responses detected in the blood on PID 21 also correlated with the responses found in the gut associated lymphoid tissues. The ASC and serum antibody responses after the challenge corresponded to a secondary immune response in the groups inoculated with attenuated virus, whereas a primary response was evident in the control group. No increase was seen in any of the parameters studied in the pigs inoculated with virulent PEDV. PMID- 11867171 TI - Effect of sulphadiazine and trimethoprim on the immune response of rainbow trout (Oncorhynchus mykiss). AB - A combination of sulphadiazine and trimethoprim at a ratio of 5:1 (SDZ/TMP) was tested for possible immunomodulatory effects. The aim of the study was to follow the immune response after vaccination with simultaneous drug treatment. The fish were immunised with a commercial oil-based divalent (furunculosis/vibriosis) vaccine and were simultaneously given oral drug treatment. The specific immune response was monitored by analysing the levels of specific antibodies with ELISA. As indicators of the nonspecific immune response, the lysozyme activity of serum was measured and the phagocytic activity of circulating leucocytes was monitored by a chemiluminescence assay. Total circulating leucocyte counts and differentials were also monitored. The disease resistance was evaluated by challenge tests at the end of the experiment. The results indicate that SDZ/TMP at a ratio of 5:1 does not interfere negatively with the immune response in rainbow trout after vaccination. A slight stimulation in the antibody response as well as in the chemiluminescence response of circulating granulocytes was recorded in fish treated with the drugs in connection with vaccination. The drugs did not significantly affect the survival after challenge. PMID- 11867173 TI - Hecker R, Sherlock S. Electrolyte and circulatory changes in terminal liver failure [Lancet 1956;2:1221-1225]. PMID- 11867174 TI - Hepatic and extrahepatic malignancies in primary sclerosing cholangitis. AB - BACKGROUND/AIMS: To assess the risk of hepatic and extrahepatic malignancies in a large cohort of Swedish primary sclerosing cholangitis (PSC) patients compared with that of the general Swedish population. METHODS: The study cohort comprised 604 PSC patients identified between 1970 and 1998. Follow-up was provided through linkages to the Swedish Cancer and Death registries. Cumulative incidence of malignancies and standard incidence ratio were calculated with the incidence rates in the Swedish population, taking into account: sex, age and calendar year as comparison group. RESULTS: Median time of follow-up was 5.7 years (range 0 27.8). Seventy-nine percent had concomitant inflammatory bowel disease. The cause of death was cancer in 44%. The frequency of hepatobiliary malignancies was 13.3% (81/604). Thirty-seven percent (30/81) of all hepatobiliary malignancies were diagnosed less than 1 year after the diagnosis of PSC. The risk for hepatobiliary malignancy was increased 161 times, for colorectal carcinoma 10 times and for pancreatic carcinoma 14 times, compared with that of the general population. CONCLUSIONS: In this national-based study including the largest cohort of PSC patients ever presented, the frequency of cholangiocarcinoma is 13%. The risk of hepatobiliary carcinoma is constant after the first year after PSC diagnosis with an incidence rate of 1.5% per year. The risk of pancreatic carcinoma is increased 14 times compared with the general Swedish population. These results are suggestive of an increased risk of pancreatic carcinoma in patients with PSC. PMID- 11867175 TI - Contribution of the two Gs-coupled PGE2-receptors EP2-receptor and EP4-receptor to the inhibition by PGE2 of the LPS-induced TNFalpha-formation in Kupffer cells from EP2-or EP4-receptor-deficient mice. Pivotal role for the EP4-receptor in wild type Kupffer cells. AB - BACKGROUND/AIMS: Prostaglandin E2 (PGE2) is known to inhibit the lipopolysaccharide (LPS)-induced tumor necrosis factor alpha (TNFalpha) formation in Kupffer cells via an increase in cAMP. Four receptor-subtypes have been cloned for PGE2 so far. Two of them, the EP2-receptor and the EP4-receptor are linked to stimulatory Gs-proteins and could mediate the inhibition by PGE2 of TNFalpha formation. METHODS: The significance of both receptors for PGE2-dependent inhibition of LPS-induced TNFalpha-formation was studied using Kupffer cells of mice in which either one of the two receptors had been eliminated by homologous recombination. RESULTS: The mRNAs of both receptors were expressed in wild type mouse Kupffer cells. Exogenous PGE2 inhibited TNFalpha-formation in Kupffer cells lacking either EP2-receptor or EP4-receptor to a similar extent as in control cells, however, 10-fold higher PGE2 concentrations were needed for half maximal inhibition in cells lacking the EP4-receptor than in control or EP2-receptor deficient cells. The response to endogenous PGE2 was blunted in EP4-receptor deficient mice only and especially after prolonged incubation. CONCLUSIONS: The data indicate, that PGE2 can inhibit TNFalpha-formation via both the EP2- and the EP4-receptor and that, however, the EP4-receptor appears to be physiologically more relevant in Kupffer cells since it conferred a high affinity response to PGE2. PMID- 11867176 TI - Bilirubin directly disrupts membrane lipid polarity and fluidity, protein order, and redox status in rat mitochondria. AB - BACKGROUND/AIMS: Unconjugated bilirubin (UCB) impairs crucial aspects of cell function and induces apoptosis in primary cultured neurones. While mechanisms of cytotoxicity begin to unfold, mitochondria appear as potential primary targets. METHODS: We used electron paramagnetic resonance spectroscopy analysis of isolated rat mitochondria to test the hypothesis that UCB physically interacts with mitochondria to induce structural membrane perturbation, leading to increased permeability, and subsequent release of apoptotic factors. RESULTS: Our data demonstrate profound changes on mitochondrial membrane properties during incubation with UCB, including modified membrane lipid polarity and fluidity (P<0.01), as well as disrupted protein mobility (P<0.001). Consistent with increased permeability, cytochrome c was released from the intermembrane space (P<0.01), perhaps uncoupling the respiratory chain and further increasing oxidative stress (P<0.01). Both ursodeoxycholate, a mitochondrial-membrane stabilising agent, and cyclosporine A, an inhibitor of the permeability transition, almost completely abrogated UCB-induced perturbation. CONCLUSIONS: UCB directly interacts with mitochondria influencing membrane lipid and protein properties, redox status, and cytochrome c content. Thus, apoptosis induced by UCB may be mediated, at least in part, by physical perturbation of the mitochondrial membrane. These novel findings should ultimately prove useful to our evolving understanding of UCB cytotoxicity. PMID- 11867177 TI - Polymorphonuclear neutrophils are a source of hepatocyte growth factor in patients with severe alcoholic hepatitis. AB - BACKGROUND/AIMS: Hepatocyte growth factor (HGF) is a pleiotropic cytokine involved in liver regeneration. Plasma HGF levels correlate with survival and hepatocyte proliferation in alcoholic hepatitis (AH). As AH is accompanied by inflammation, neutrophilia and polymorphonuclear neutrophil (PMN) infiltration of the liver, we postulated that PMN could be a source of HGF in such patients. METHODS: We studied 25 patients with severe AH in comparison with 20 alcoholic cirrhotic patients without AH and 20 healthy controls; the impact of a 28-day course of corticosteroids was evaluated in patients with AH. RESULTS: On day 0, HGF plasma and homogenized liver tissue levels were markedly increased in AH patients as compared to controls. The role of PMN in HGF production during AH was confirmed by a significantly higher ex-vivo HGF production capacity of lipopolysaccharide-stimulated blood PMN from AH patients relative to both control groups. Formyl-Methionyl-Leucyl-Phenylalanine-induced PMN release of HGF (degranulation conditions) was also higher in AH patients. In this setting, we found that HGF release by PMN ex vivo correlated strongly with HGF plasma levels, and that the degree of hepatic PMN correlated strongly with hepatic HGF levels. HGF plasma levels and ex-vivo HGF release by PMN were unaffected by steroid therapy. CONCLUSIONS: These findings suggest that, by releasing HGF, PMN could participate in liver regeneration during severe alcoholic hepatitis. PMID- 11867178 TI - Increased plasma levels of neuropeptide Y in hepatorenal syndrome. AB - BACKGROUND/AIMS: To investigate the relationship between neuropeptide Y (NPY), a potent renal vasoconstrictor peptide released upon marked stimulations of sympathetic nervous system (SNS), and renal and circulatory function in cirrhosis. METHODS: Plasma levels of NPY (radioimmunoassay) and norepinephrine and renal function parameters were determined in 17 healthy controls, nine patients with cirrhosis without ascites, and 37 patients with cirrhosis and ascites, of whom 12 had hepatorenal syndrome (HRS). RESULTS: Patients with ascites showed circulating levels of NPY similar to those of patients without ascites and controls (73+/-4, +/-4 and 68+/-4 pmol/l, respectively; NS). However, patients with HRS had significantly increased levels of NPY with respect to the other groups (110+/-6 pmol/l; P<0.001). NPY levels correlated inversely with renal plasma flow and glomerular filtration rate and directly with norepinephrine. In patients with HRS (n=6) treatment with terlipressin and albumin was associated with a marked improvement in circulatory and renal function and marked suppression of NPY and norepinephrine levels. CONCLUSIONS: Patients with HRS have increased levels of NPY which are related to circulatory dysfunction and SNS activation and may contribute to renal vasoconstriction. PMID- 11867179 TI - Total effective vascular compliance in patients with cirrhosis. Effects of propranolol. AB - BACKGROUND: Total effective vascular compliance (TEVC), may be increased in cirrhosis. However, its significance is unclear. AIMS: To investigate TEVC in patients with cirrhosis, and the effects of propranolol. METHODS: Seven patients without liver disease and 44 cirrhotic patients were studied before and after double-blind administration of propranolol (n=33) or placebo (n=11). MEASUREMENTS: TEVC (right atrial pressure response to rapid central volume expansion), hepatic venous pressure gradient (HVPG) and systemic hemodynamics. RESULTS: TEVC (ml x mmHg(-1) x kg(-1)) was increased in cirrhotics (1.67 +/- 0.66 versus 1.33 +/- 0.32 in controls; P<0.05). TEVC was not modified by placebo, but was markedly reduced by propranolol (from 1.74 +/- 0.75 to 1.33 +/- 0.56; P<0.01). Propranolol decreased HVPG >10% in 20 patients ('responders': -20 +/- 9%) but <10% in 13 'non-responders'. TEVC was normalized by propranolol in HVPG 'responders' (from 1.76 +/- 0.88 to 1.21 +/- 0.51; P<0.01), but not in 'non responders' (1.69 +/- 0.48 to 1.52 +/- 0.59; NS). Reduction of TEVC in responders was accompanied by increased free hepatic vein pressure (+21 +/- 20%, P=0.05; approximately 60% of the fall in HVPG), which was not observed in non-responders (+3 +/- 11%, NS). CONCLUSIONS: TEVC is increased in cirrhosis. This abnormality is corrected by propranolol in patients exhibiting a >10% fall in HVPG, suggesting that changes in vascular compliance may influence the portal pressure response to propranolol. PMID- 11867180 TI - Regulation of hepatic stellate cell proliferation and collagen synthesis by proteinase-activated receptors. AB - BACKGROUND/AIMS: Thrombin and MC tryptase, which are agonists for proteinase activated receptors-1 and -2, respectively, are both increased in injured liver. We have examined if rat stellate cells express these receptors and if receptor agonists influence stellate cell activation. METHODS: Expression of mRNA for proteinase activated receptors-1 and -2 were examined by RT-PCR and Northern blotting in lysates of cultured stellate cells and receptor protein examined by Western blotting. The effects of receptor agonists on cell proliferation and collagen synthesis were examined by 3H-thymidine and 3H-proline incorporation assays, respectively. RESULTS: Rat stellate cells activated by culture on plastic showed a progressive increase in expression of proteinase-activated receptor-1 and -2 mRNA and proteinase-activated receptor-2 protein as they transformed to a myofibroblastic phenotype. Proteinase-activated receptor-1 agonists thrombin and the peptide SFFLRN, and proteinase-activated receptor-2 agonists tryptase and the peptide SLIGRL induced stellate cell proliferation and the rapid phosphorylation of 44 and 42 kDa mitogen-activated protein kinases. PD98059, an inhibitor of these kinases, inhibited this proliferative response. Both tryptase and SLIGRL increased collagen secretion by stellate cells. CONCLUSIONS: This study indicates that the natural proteinase-activated receptor agonists thrombin and MC tryptase might sustain liver fibrosis by promoting stellate cell proliferation and collagen synthesis. PMID- 11867181 TI - Pyrrolidine dithiocarbamate protects against thioacetamide-induced fulminant hepatic failure in rats. AB - BACKGROUND/AIMS: Reactive oxygen species and nuclear factor kappa B (NF-kappaB) activation have been implicated in the pathogenesis of cell injury in experimental models of liver damage. The aim of the present study was to examine whether pyrrolidine dithiocarbamate (PDTC), an anti oxidant and inhibitor of NF kappaB activation, would prevent hepatic damage induced in a rat model of thioacetamide (TAA)-induced liver failure. METHODS: Fulminant hepatic failure was induced in the control and treatment groups by two intraperitoneal injections of TAA (either 300 or 400 mg/kg) at 24-h intervals. In the treatment groups, rats were treated also with PDTC (60 mg/kg/24 h, i.p.), initiated 24 h prior to TAA. RESULTS: Liver enzymes, blood ammonia, and hepatic levels of thiobarbituric acid reactive substances (P<0.001) and protein carbonyls (P<0.05) were significantly lower in rats treated with PDTC compared to TAA only. Liver histology and the survival rate in the PDTC-treated rats were also improved (P<0.01 compared to TAA only). NF-kappaB activation, 2 and 6 h after TAA administration, was inhibited by PDTC. CONCLUSIONS: In a rat model of fulminant hepatic failure, the administration of PDTC attenuated liver damage and improved survival. This effect may be due to decreased oxidative stress and inhibition of NF-kappaB activation. PMID- 11867182 TI - Activation of signal transducer and activator transcription 3 and expression of suppressor of cytokine signal 1 during liver regeneration in rats. AB - BACKGROUND/AIMS: Recent work has shown that the signal transducer and activator of transcription (STAT) 3 activation is important for the initiation of the proliferative response following partial hepatectomy (PH). However, the issue of where STAT3 is activated and how it is regulated is unclear. The aims of this study were to identify STAT3-activated cells and to clarify the expression of suppressor of cytokine signal (SOCS), a negative feedback molecule of STAT3, after PH. METHODS: STAT3-activated cells and SOCS1-positive cells were identified by immunohistochemistry after a two-thirds PH in rats. SOCS mRNA was examined by Northern analysis. RESULTS: STAT3 was activated in hepatocytes from those localized in the periportal zones of hepatic lobules after PH. STAT3 activation was also detected in Kupffer cells and sinusoidal endothelial cells prior to its detection in hepatocytes. After STAT3 activation, SOCS1 protein in response to PH was detected immunohistochemically in regenerating liver. SOCS1 and SOCS3 mRNA were induced in regenerating liver after PH. CONCLUSIONS: STAT3 signaling can occur in Kupffer cells and sinusoidal endothelial cells prior to in hepatocytes from those localized in the periportal zones. SOCS1 as well as SOCS3 may regulate STAT3 signaling negatively after PH. PMID- 11867184 TI - The presence and the role of chromatin cholesterol in rat liver regeneration. AB - BACKGROUND/AIMS: It has been shown that cholesterol is necessary in early G1 phase during cell duplication. In the present research we have studied the presence of cholesterol in the hepatocyte chromatin lipid fraction and its behaviour in liver regeneration. METHODS: Hepatocyte nuclei and chromatin were isolated from normal and regenerating rat liver. The lipid fraction was extracted and analysed by chromatography. The activity of sphingomyelin-synthase in the chromatin was evaluated using labelled phosphatidylcholine. RESULTS: In the chromatin, the amount of cholesterol is similar to that of sphingomyelin, and it increases in chromatin digested with exogenous sphingomyelinase or proteinase K. It may be concluded that a complex, formed by cholesterol, sphingomyelin and proteins, is present in the chromatin. The particular affinity between sphingomyelin and cholesterol in chromatin with respect the nuclear membrane may be tentatively explained as due to the enrichment in saturated fatty-acids of the chromatin sphingomyelin. Moreover the cholesterol inhibits the chromatin sphingomyelin-synthase activity. During liver regeneration, an increase in chromatin cholesterol is observed between 6 and 18 h after hepatectomy, when the neutral-sphingomyelinase activity increases and the sphingomyelin-synthase is inhibited. CONCLUSIONS: The cholesterol is present in the chromatin and its amount changes in relation to cell proliferation in regenerating liver. PMID- 11867183 TI - Overexpression of the mouse Fas gene in human Hep3B hepatoma cells overcomes their resistance to Fas-mediated apoptosis. AB - BACKGROUND/AIMS: Fas-induced apoptosis is one of the main forms of apoptosis occurring in hepatocytes. We have previously demonstrated that the human hepatoma cell line Hep3B is resistant to Fas-mediated apoptosis. In this study, we investigated whether the human Fas receptor itself, or the Fas transduction pathway was responsible for the resistant phenotype. METHODS: Clones of Hep3B cells overexpressing the mouse Fas gene (Hep3B(mfas)) were generated by transfection, and apoptosis was studied by (i) chromatin condensation and nuclear fragmentation, (ii) flow cytometry, (iii) DNA fragmentation and (iv) poly (ADP ribose) polymerase cleavage. RESULTS: Use of the species-specific and agonistic anti-mFas monoclonal antibody (JO2), showed that the mFas receptor was correctly routed to the plasma membrane of Hep3B(mfas) cells. Using the four above mentioned criteria, we demonstrated that JO2 triggered mFas-mediated apoptosis of Hep3B(mfas), but not of Hep3B(pCi) cells (transfected with an empty vector). CONCLUSIONS: Our data show (i) that the Fas signaling pathway can be completed when a functional mFas receptor is expressed in Hep3B cells, and thus, (ii) that the death-inducing signaling complex components and the effector caspases are functional in Hep3B cells. Moreover, they suggest that the Fas subunits are not pre-assembled at the cell membrane before receptor-ligand interaction. PMID- 11867185 TI - Emotional distress in chronic hepatitis C patients not receiving antiviral therapy. AB - BACKGROUND/AIMS: The aim of our study was to determine the prevalence, type, and severity of emotional distress in a large group of consecutive chronic hepatitis C (CHC) patients not receiving anti-viral therapy. METHODS: The brief symptom inventory and a 67-item questionnaire with the SF-36 embedded within it were used to study 220 outpatients with compensated CHC. RESULTS: Seventy-seven (35%) participants reported significantly elevated global severity index (GSI) T-scores compared to an expected frequency of 10% in population controls. In addition, significantly elevated depression, anxiety, somatization, psychoticism, and obsessive-compulsive subscale T-scores were reported in 28-40% of subjects. Subjects with an active psychiatric co-morbidity had significantly higher GSI and subscale T-scores compared to subjects with active medical co-morbidities and subjects without medical or psychiatric co-morbidities (P<0.01). However, patients with CHC alone also had a higher frequency of elevated GSI T-scores compared to population controls (20 versus 10%). GSI and subscale T-scores were strongly associated with SF-36 summary scores (P<0.001). CONCLUSIONS: Clinically significant emotional distress was reported in 35% of CHC patients not receiving antiviral therapy. In addition to depression, a broad array of psychological symptoms were observed. Further investigation into the etiopathogenesis and treatment of emotional distress in CHC patients is warranted. PMID- 11867186 TI - Features of the CD4+ T-cell response in liver and peripheral blood of hepatitis C virus-infected patients with persistently normal and abnormal alanine aminotransferase levels. AB - BACKGROUND/AIMS: The liver is the primary site of hepatitis C virus (HCV) replication; intrahepatic T-cell responses may influence liver disease severity. METHODS: HCV-specific CD4(+) T-cell reactivity was investigated ex vivo in paired liver tissue and peripheral blood from 42 chronic HCV patients. RESULTS: The frequencies with which HCV-specific HLA class-II-restricted CD4(+) T-cell proliferation were observed were 29% in liver and 36% in peripheral blood. Among responses, non-structural-3 protein (NS3)-specific T-cell proliferation was dominant but non-exclusive and did rarely occur concurrently in liver infiltrate and peripheral blood suggesting liver compartmentalization of a CD4(+) T-cells population. Compared with 24 patients with abnormal ALT levels, 18 HCV carriers with persistently normal ALT levels had similar serum and liver viral loads but showed: (i) a low-activity grade and stage chronic hepatitis (P<0.001); (ii) less intrahepatic CD4(+) T-lymphocytes (P<0.01); (iii) less frequent intrahepatic (17 vs. 33%) and peripheral (17 vs. 38%) NS3-specific CD4(+) T-cell proliferation; (iv) less often in vitro T-helper (Th)1 (interferon-gamma) cytokine production (2 vs. 18%; P<0.001). CONCLUSIONS: Our data show a low frequency of intrahepatic HCV specific HLA class-II-restricted CD4(+) Th1 responses in patients with chronic HCV. However, these Th1 responses are detected more often in those patients with overt clinical and histological disease. PMID- 11867187 TI - Human and swine hepatitis E viruses from Western India belong to different genotypes. AB - BACKGROUND/AIMS: Hepatitis E is endemic in India. Earlier, we showed prevalence of IgG antibodies to hepatitis E virus (IgG-anti-HEV) in different animal species and inability of at least one human hepatitis E virus (HEV) strain to infect pigs. In the US where hepatitis E is not endemic in humans, zoonotic spread of HEV was suspected as swine and human HEV were closely related and cross-species infection was documented. The present study attempts to identify and partially characterize swine HEV from India. METHODS: Serum samples from 284 pigs were screened for the presence of HEV-RNA (nested polymerase chain reaction; PCR) and IgG-anti-HEV (enzyme-linked immunosorbent assay; ELISA). PCR products (Open Reading Frame-2 region) were sequenced and subjected to phylogenetic analysis. Two sero-negative pigs were inoculated with swine HEV-positive serum pool. RESULTS: ELISA and PCR positivity were 42.9 and 4.6%, respectively. All Indian swine HEV sequences clustered with genotype IV. Pigs could be experimentally infected with swine HEV. CONCLUSIONS: Swine HEV circulates in Indian pigs. In contrast to US and Taiwan wherein both human and swine HEV isolates belong to same genotype, Indian human HEV isolates belong to genotype I whereas genotype IV circulates in swine. Though experimental infection with Indian swine HEV was possible, at least one human HEV strain could not infect pigs. PMID- 11867188 TI - Genomic heterogeneity of hepatitis B virus (HBV) and outcome of perinatal HBV infection. AB - BACKGROUND/AIMS: Data regarding hepatitis B virus (HBV) genomic heterogeneity in perinatal infection are incomplete, although HBV variants might be involved in neonatal fulminant hepatitis (ALF). We investigated HBV variability in infected babies showing different clinical courses. METHODS: We analyzed HBV genomes isolated from nine vertically infected babies and the mothers of four of them. Two infants born to HBe-antigen (HBeAg)-positive women developed a chronic infection; seven babies (six born to anti-HBe mothers) developed acute hepatitis that had a fulminant course in four cases and a benign course in three. Two babies developing ALF received anti-HBV immunoprophylaxis at birth. RESULTS: Viruses carrying no significant mutation infected infants born to HBeAg-positive women. HBeAg-defective viruses were detected both in children with benign and fulminant hepatitis and their mothers. A double nucleotide mutation at positions 1762 and 1764 of the HBV core-promoter was found in two of the four infants with ALF, although it was not detected in isolates from the mother of one of them. No significant S gene mutation was found in HBV from any of the babies. CONCLUSIONS: This study indicates that HBV genomic heterogeneity is not primarily involved either in the evolution of the infection or the failure of neonatal HBV immunoprophylaxis. PMID- 11867189 TI - Primary sclerosing cholangitis: neoplastic potential in bile ducts, colon and the pancreas? PMID- 11867190 TI - Reduced quality of life in hepatitis C--is it all in the head? PMID- 11867191 TI - Benign recurrent intrahepatic cholestasis progressing to progressive familial intrahepatic cholestasis: low GGT cholestasis is a clinical continuum. AB - Benign recurrent intrahepatic cholestasis (BRIC) is an autosomal recessive liver disease, characterised by intermittent attacks of cholestasis, which can start at any age and last for several weeks to months. Characteristically serum GGT activity is low and normal liver structure is preserved. Progressive familial intrahepatic cholestasis (PFIC) is another liver disease, characterised by severe cholestasis, starting almost invariably before 6 months of age. All patients progress to cirrhosis, liver failure and death, unless a liver transplantation is performed. We now identified four patients who presented in childhood with recurrent attacks of cholestasis, while in the course of the disease the cholestasis gradually became permanent. Although liver biopsies performed in the early stages of the disease showed normal liver architecture, late stage biopsies revealed evident fibrosis with porto-portal septa formation. In conclusion, the disease of these patients started with the clinical and histopathological characteristics of BRIC but progressed to PFIC. PMID- 11867192 TI - Spontaneous bacterial arthritis in a cirrhotic patient. PMID- 11867193 TI - Is hepatitis B virus subtype testing useful in predicting virological response and resistance to lamivudine? PMID- 11867194 TI - A long-awaited small animal model for hepatitis C. PMID- 11867195 TI - Chemopreventive effects of Aloe arborescens on N-nitrosobis(2-oxopropyl)amine induced pancreatic carcinogenesis in hamsters. AB - The modification effects of freeze-dried aloe (Aloe arborescens) whole leaf powder during the initiation phase of carcinogenesis were investigated in hamsters treated with N-nitrosobis(2-oxopropyl)amine (BOP). Female Syrian hamsters were given four weekly subcutaneous injections of BOP at a dose of 10mg/kg and then given 0, 1 or 5% aloe in their diet for 5 weeks. At week 54 of the experiment, all surviving animals were sacrificed and development of neoplastic and preneoplastic lesions was assessed histopathologically. The incidences of pancreatic adenocarcinomas, atypical hyperplasias or total atypical hyperplasias plus adenocarcinomas were significantly (P<0.05) decreased with BOP+5% aloe, and that of adenocarcinomas were also significantly (P<0.05) reduced in the BOP+1% aloe as compared to the BOP alone group. Multiplicities of pancreatic adenocarcinomas, atypical hyperplasias or total lesions were also significantly (P<0.01 or P<0.05) lower in the BOP+5% aloe group than with the BOP alone. Quantitative data for neoplastic lesions in the lung, liver, gall bladder, kidney and urinary bladder of hamsters were not significantly different among the three groups. In a satellite experiment, pretreatment with aloe significantly (P<0.01) reduced the formation of O6-methyldeoxyguanosine in epithelial cells of pancreatic ducts as compared to the BOP alone value. Our results thus indicate that aloe prevents BOP-induced pancreatic neoplasia in hamsters in relation to decreased DNA adduct formation in the target tissue. PMID- 11867196 TI - Serum retinol and risk of hepatocellular carcinoma in patients with child-Pugh class A cirrhosis. AB - Liver cirrhosis is the main risk factor for the development of human hepatocellular carcinoma (HCC). In this condition, the liver and plasma suffer a drastic depletion of retinoids. This study was conducted to investigate whether any relation exists between serum retinol levels and HCC development in cirrhotics. Seventy child-Pugh class A cirrhotic patients, 16 child-Pugh class A cirrhotic patients with HCC, and 140 age- and sex-matched subjects were included in this study. At the time of enrollment, fasting blood samples were taken to determine serum retinol levels. For the following 7 years, the 70 cirrhotic patients were also followed up for the occurrence of HCC by periodic screening with ultrasonography and serum alpha-fetoprotein assays. The serum retinol levels in both cirrhotic patients and HCC patients were significantly lower than those in healthy subjects. Among the 70 cirrhotic patients, 14 HCC were detected during follow-up. The prediagnostic retinol levels were significantly lower in cirrhotic patients who developed HCC compared with patients who did not. The odds ratio of cirrhotic patients who developed HCC in the lowest tertile to highest tertile of retinol status was 6.75 (95% CI=1.26--36.0; P=0.015). Our results suggest that a state of retinoid deficiency may promote hepatocarcinogenesis in patients at high risk such as cirrhotics. PMID- 11867197 TI - Inhibition of conjugated fatty acids derived from safflower or perilla oil of induction and development of mammary tumors in rats induced by 2-amino-1-methyl-6 phenylimidazo[4,5-b]pyridine (PhIP). AB - Chemopreventive effects of conjugated fatty acids derived from safflower oil (CFA S), which contains large amounts of conjugated linoleic acid, and from perilla oil (CFA-P) with abundant conjugated alpha-linolenic acid were examined in a 2 amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP)-induced rat mammary carcinogenesis model. Groups of 20-22 6-week-old female Sprague-Dawley (SD) rats were given eight intragastric injections of PhIP at a dose of 100 mg/kg b.w. during the initial 8 week period. Powdered basal diets containing 0.1% CFA-S or CFA-P were applied during or after PhIP treatment until week 40. In the rats receiving CFA-S or CFA-P together with PhIP treatment, retardation of mammary tumor emergence was observed until week 27. The groups given CFA-S or CFA-P after PhIP treatment, in contrast, demonstrated significant decrease in the final incidences of mammary adenocarcinomas. The indices of proliferating cell nuclear antigen positive cells in mammary adenocarcinomas were significantly reduced with both CFA-S and CFA-P in the post-initiation phase. Formation of aberrant crypt foci in the colon and basophilic foci of the pancreas due to the PhIP treatment group were not affected by CFA-S or CFA-P. In a second short-term experiment, female SD rats were maintained on powdered basal diet containing 0.03% PhIP alone or together with 0.1% CFA-S or CFA-P for 4 weeks. Immunohistochemically, CFA-S and CFA-P were revealed to suppress PhIP-DNA adduct formation in the epithelial cells of mammary gland (duct and alveolar cells), colon and pancreas. These results indicated that CFA-P and CFA-S may retard development of PhIP-induced mammary tumors with inhibition of PhIP-DNA adduct formation, and decreased mammary carcinogenesis in the post-initiation period with inhibition of cell proliferation. PMID- 11867198 TI - Comparison of idarubicin and daunorubicin regarding intracellular uptake, induction of apoptosis, and resistance. AB - Anthracycline antibiotics are widely used as anticancer agents. Idarubicin (4 demethoxydaunorubicin; Ida), a semisynthetic derivative of daunorubicin (Dnr) is more potent than the parent compound in vitro and in vivo. The equitoxic dose of Ida in patients is about one-fourth of that of Dnr. We compared these drugs regarding cytotoxicity, apoptosis induction, and resistance mechanisms in human leukaemic cell lines. Cytotoxicity was studied by means of the 3-[4,5 dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide assay and drug-induced apoptosis by means of the Annexin V-fluorescein isothiocyanate method at similar intracellular concentrations (extracellular concentrations of 0.35 microM for Ida and 1 microM for Dnr). Ida was at least twice as potent as Dnr in MOLT-4, HL60, CEM, and K562 cell lines. It took 8 h for Ida to induce approximately 20% apoptosis, but at least 22 h for Dnr to reach 20% apoptosis at identical intracellular concentration. Ida induces a faster and higher apoptosis rate compared with Dnr. The human chronic myelogenous leukaemia cell line (K562) was selected for resistance to Dnr and Ida with and without verapamil (Ver). Continuous incubation with Dnr, but not with Ida, led to an increased mdr1 gene expression as assessed by real-time PCR. The development of mdr1 gene expression in Dnr-resistant cells could be reversed by the presence of Ver. Ver also reversed the cytotoxicity to Dnr, but not to Ida, in K562/Dnr cells. The results show that Ida is more effective than Dnr in inducing apoptosis and that there are differences in resistance mechanisms between the drugs. PMID- 11867200 TI - Dibenzoylmethane induces cell cycle deregulation in human prostate cancer cells. AB - Dibenzoylmethane (DBM), a minor beta-diketone constituent of licorice and sunscreens, has been shown to exhibit anti-neoplastic effects in chemically induced skin and mammary cancers in several animal models. To date, no mechanism for the growth inhibitory effects of DBM on prostate cancer cells has been proposed. In this study, we examined the effects of DBM on the growth and cell cycle kinetics of several human prostate carcinoma cell lines. Using an MTT cytotoxicity assay, IC50 values of 25-100 microM were observed following 72 h exposure to DBM. LNCaP, DU145, and PC-3 prostate carcinoma cell lines were particularly sensitive in comparison to the cells with the vehicle alone. Flow cytometric analyses showed deregulation of the cell cycle, which correlated with the observed cytostatic effects of DBM in prostate carcinoma cells. These data suggest a potential role for DBM in the prevention and treatment of prostate cancer. PMID- 11867199 TI - Reduction of in vivo tumor growth by MMI-166, a selective matrix metalloproteinase inhibitor, through inhibition of tumor angiogenesis in squamous cell carcinoma cell lines of head and neck. AB - Matrix metalloproteinases (MMPs) have been implicated in tumor invasion, metastasis, and angiogenesis. We have recently shown that MMI-166, a new orally active MMP inhibitor specific for MMP-2 and -9, suppressed experimental metastasis of Lewis lung cancer, C-H1 human colon cancer, and pancreatic cancer without affecting tumor growth in vitro. In the present study, we determined whether oral administration of MMI-166 reduces tumor growth not only in such tumors but also in squamous cell carcinoma of head and neck (SCCHN). MMI-166 inhibited both activity of MMP-2 and -9 without affecting steady state levels of their mRNAs in SCCHN. Interestingly, protein levels of MMP-2 and -9 from the cultures were drastically diminished by culturing with MMI-166. This was also observed in xenografts of MMI-166-administered mice. In addition, daily oral administration of MMI-166 (100mg/kg) inhibited local tumor growth accompanied by the reduction of blood vessel density and Ki-67-positivity and increase in terminal deoxynucleotidyl transferase-mediated cUDP nick-end labeling (TUNEL) positivity. These results suggested that orally administered MMI-166 reduced in vivo tumor growth of SCCHN through inhibition of angiogenesis and induction of apoptosis accompanied by the reduction of MMP productions and activities. Therefore, MMI-166 seems to be useful for tumor dormancy therapy of SCCHN. PMID- 11867201 TI - Apoptosis and expression of its regulatory proteins in soft tissue sarcomas. AB - Information for the occurrence and extent of apoptosis in soft tissue sarcoma (STS) and their clinical implication are limited. In 102 cases of STSs, apoptosis was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling method and expression of Bcl-2, Bax, MIB-1 and p53 protein was examined immunohistochemically. The apoptotic index of the STSs ranged from 0 to 15% with an average value of 1.9%. The mean values of positive cell staining for Bcl-2, Bax, MIB-1 and p53 protein were 32.1, 40.8, 17.0 and 20.3%, respectively. The extent of apoptosis and expression of Bcl-2 protein were correlated to the histologic types of the tumor. Synovial sarcoma had a significantly higher expression of Bcl-2 protein, and lower incidence of apoptosis. STS underwent apoptosis at a constitutional level. There were no significant relationships between extent of the apoptosis, expression of its regulatory proteins and prognosis of the patients. PMID- 11867202 TI - Polymorphisms in the estrogen receptor-alpha gene and breast cancer risk. AB - The estrogen receptor-alpha (ERalpha) has been known to play a role in the development and progression of breast cancer. Several genetic polymorphisms in the ERalpha gene have been related to breast cancer risk and/or different tumor characteristics. In this study, PCR and direct sequencing based methods were used to examine this issue further in a Korean study population consisting of 155 women, 110 with breast cancer and 45 without cancer. We also assessed the potential role of the ERalpha genotype in ER, PR, p53, c-erbB2, and bcl-2 expression. Only one of the allelic variants of ERalpha gene was found in our study subjects; the (C(975)G) change was present in half of the study subjects. Although this allele had no direct effect in individual breast cancer risk, it was positively associated with tumor PR (P for trend=0.04) and ER expression (P for trend=0.06) and negatively associated with p53 expression (P for trend=0.02). PMID- 11867203 TI - XRCC1 polymorphisms and head and neck cancer. AB - Inter-individual differences in DNA repair capacity have been demonstrated using a variety of phenotypic assays, including reduced repair among patients with squamous cell carcinoma of the head and neck (SCCHN). The XRCC1 DNA repair gene may facilitate DNA strand break and base excision repair. A recent case-control study of SCCHN reported associations with two polymorphisms of the XRCC1 including the exon 6, 194Arg/Arg genotype and the exon 10, 399 Gln/Gln genotype. We conducted an analysis of these two XRCC1 polymorphisms using data from a case control study of SCCHN. Among white subjects, we found a weak elevation in risk associated with the Arg194Trp polymorphism [odds ratio (OR)=1.3; 95% confidence interval (CI)=0.6-2.9] and a decreased risk for the Arg399Gln polymorphism (OR=0.6; CI=0.4-1.1). We found a markedly decreased odds ratio for the Gln/Gln genotype among whites (OR=0.1; CI=0.04-0.6) and blacks (OR=0.01; CI=0.0004-0.3). We also found a suggestion of an interaction between the Arg194Trp and Arg399Gln polymorphisms and tobacco use. Additional epidemiologic and functional studies are needed to resolve the importance of these XRCC1 polymorphisms in SCCHN. PMID- 11867204 TI - Telomerase overexpression in K562 leukemia cells protects against apoptosis by serum deprivation and double-stranded DNA break inducing agents, but not against DNA synthesis inhibitors. AB - Telomeres are specialized DNA/protein structures that act as protective caps to prevent end fusions. The maintenance of telomeres is essential for chromosomal stability. Telomerase is regulated by human telomerase reverse transcriptase (hTERT). c-Myc oncoprotein is also implicated in the positive regulation of hTERT expression. We show here that two clones of hTERT-transfected K562 erythroleukemia cells have elongated telomeres (22.5 and 24.0 kb), whereas telomere length of both c-Myc-transfected K562 cells and parental K562 cells is 6.5 kb. Telomerase activity and hTERT mRNA expression increased in hTERT transfected K562 cells, while the expression levels of telomerase activity and hTERT in c-Myc-transfected K562 cells were similar to that in parental K562 cells, despite an overexpression of c-Myc. Importantly, we found that hTERT transfected K562 cells are protected against apoptosis induced by serum deprivation and double-stranded DNA break inducing agents (ionizing irradiation, and etoposide (VP-16)), but not against DNA synthesis inhibitors (1-beta-D arabinofuranosylcytosine and hydroxyurea). These findings suggest that overexpression of telomerase by transfecting hTERT confers telomere-elongation and resistance to double-stranded DNA break inducing agents. PMID- 11867205 TI - Genetic analysis of the liver putative tumor suppressor (LPTS) gene in hepatocellular carcinomas. AB - Recently, a novel liver-related putative tumor suppressor (LPTS), which has a growth inhibitory function in the hepatocellular carcinoma (HCC) cell line, has been identified at chromosome 8p23. To determine the relationship of the LPTS with the development or progression of HCC, we analyzed the genetic alterations and the expression pattern of the LPTS gene in a series of 80 HCCs, six dysplastic nodules, and eight large regenerating nodules, determining the genomic structures. We identified a total of seven exons, of which two were alternative, and three LPTS isoforms, short (LPTS-S), medium (LPTS-M), and long-sizes (LPTS L). In the genetic alteration study of the LPTS gene, no mutation was detected in the large regenerating nodules, dysplastic nodules, and HCC, whereas ten (34.5%) of 29 informative cases at one or more intragenic polymorphic sites showed loss of heterozygosity (LOH). Interestingly, LOH was identified only in HCC samples with hepatitis B virus (HBV) infection and the frequency of LOH was not statistically related with histologic grade and clinical stage, suggesting that allelic loss of the LPTS gene may occur as an early event in the development of HCC, especially in the cases with HBV infection. PMID- 11867206 TI - The influence of combretastatin A-4 and vinblastine on interstitial fluid pressure in BT4An rat gliomas. AB - The influence of combretastatin A-4 disodium phosphate (CA-4, 50mg/kg intraperitoneally (i.p.)) and vinblastine (2mg/kg i.p.) on interstitial fluid pressure (IFP) was assessed in BT4An rat gliomas implanted subcutaneously in the neck. Furthermore the growth inhibitory effect of vinblastine and the distribution of fluorescence-conjugated vinblastine (BODIPY-vinblastine) were investigated. Tumors at different volumes were compared. Whereas CA-4 had no major influence on IFP, independent of tumor size, vinblastine increased the IFP in neoplasms above 8 cm(3) (P=0.03). Vinblastine yielded a significant tumor response only in tumors below 2.1 cm(3) (P=0.03). The distribution of BODIPY vinblastine was heterogeneous and comparable despite tumor volume differences. We conclude that the influence of vinblastine on IFP is more pronounced than that of CA-4 in BT4An neck tumors, and that vinblastine may reduce subsequent drug delivery to solid tumors by increasing the IFP. PMID- 11867207 TI - Molecular dissection of the human Y-chromosome. AB - Human Y chromosome, earlier thought to be gene deficient, has attracted a great deal of attention owing to its supremacy in male sex determination and unique haplotype status in the genome. Studies on Y chromosome have shown the presence of different types of satellite DNA and several genes implicated with a variety of physical and physiological functions. The interaction of these repetitive DNA with genes in normal individuals and in patients with Y-chromosome-related genetic anomalies is still an unresolved issue and is actively being pursued. The fast changing scenario of the human genome project is likely to effect our overall understanding of the Y chromosome and Y-linked genetic anomalies in a big way. We provide a brief overview of the organization of Y chromosome with respect to several important loci encompassing both the arms and their likely involvement/modulation in genetic anomalies. The experimental approaches discussed here are envisaged to be of clinical relevance for the molecular diagnosis of the Y-linked disorders. PMID- 11867208 TI - Molecular organization of internal telomeric sequences in Chinese hamster chromosomes. AB - In Chinese hamster extended blocks of telomeric-like repeats were previously detected by in situ hybridization at the pericentromeric region of most chromosomes and short arrays were localized at several interstitial sites. In this work, we analyzed the molecular organization of internal telomeric sequences (ITs) in the Chinese hamster genome. In genomic transfers hybridized with a telomeric probe, multiple Bal31 insensitive fragments were detected. Most of the fragments ranged in size between less than 1 kb and more than 100 kb and some were polymorphic. Fluorescence in situ hybridization experiments on DNA fibers and on elongated chromosomes showed that the pericentromeric ITs are composed of extensive and essentially continuous arrays of telomeric-like sequences. We then isolated three genomic regions which contain short ITs. These ITs are localized at interstitial sites (3q13-15, 3q21-26, 1p26) and are composed of 29-126 bp of (TTAGGG)(n) repeats. A peculiar feature of all the three ITs is the AT richness of the flanking sequences. Since AT-rich DNA is known to be unstable and characteristic of several mammalian fragile sites, we propose that the three ITs were inserted at these sites during the repair of double strand breaks. PMID- 11867209 TI - Molecular characterization and expression analysis of Mtmr2, mouse homologue of MTMR2, the Myotubularin-related 2 gene, mutated in CMT4B. AB - Charcot-Marie-Tooth type 4B (CMT4B) is caused by mutations in the myotubularin related 2 gene, MTMR2, on chromosome 11q22. To date, six loss of function mutations and one missense mutation have been demonstrated in CMT4B patients. It remains to be determined how dysfunction of a ubiquitously expressed phosphatase causes a demyelinating neuropathy. An animal model for CMT4B would provide insights into the pathogenesis of this disorder. We have therefore characterized the mouse homologue of MTMR2 by reconstructing the full-length Mtmr2 cDNA as well as the genomic structure. The 1932 nucleotide open reading frame corresponds to 15 coding exons, spanning a genomic region of approximately 55 kilobases, on mouse chromosome 9 as demonstrated by fluorescence in situ hybridization analysis. A comparison between the mouse and human genes revealed a similar genomic structure, except for the number of alternatively spliced exons in the 5' untranslated region, two in mouse and three in man. In situ hybridization analysis of mouse embryos showed that Mtmr2 was ubiquitously expressed during organogenesis at E9.5, with some areas of enriched expression. At E14.5, Mtmr2 mRNA was more abundant in the peripheral nervous system, including in dorsal root ganglia and spinal roots. PMID- 11867210 TI - Phenotypic consequences of mutations in the conserved motifs of the putative helicase domain of the human Cockayne syndrome group B gene. AB - Cockayne syndrome (CS) is a human genetic disorder characterized by several neurological and developmental abnormalities. Two genetic complementation groups, CS-A and CS-B, have been identified. The CSB protein belongs to helicase superfamily 2, and to the SWI/SNF family of proteins. The CSB protein is implicated in transcription-coupled repair (TCR), basal transcription and chromatin remodeling. In addition, CS cells undergo UV-induced apoptosis at much lower doses than normal cells. However, the molecular function of the CSB protein in these biological pathways has remained unclear. Evidence indicates that the integrity of the Walker A and B boxes (motifs I and II) are important for CSB function, but the functional significance of the helicase motifs Ia, III--IV has not been previously examined. In this study, single amino acid changes in highly conserved residues of helicase motifs Ia, III, V, VI and a second putative nucleotide-binding motif (NTB) of the CSB protein were generated by site-directed mutagenesis to analyze the genetic function of the CSB protein in survival, RNA synthesis recovery and apoptosis after UV treatment. The survival analysis of these CS-B mutant cell lines was also performed after treatment with the chemical carcinogen, 4-nitroquinoline-1-oxide (4-NQO). The lesions induced by UV light, cyclobutane pyrimidine dimers, are known to be repaired by TCR whereas the lesions induced by 4-NQO are repaired by global genome repair. The results of this study demonstrate that the point mutations in highly conserved residues of helicase motifs Ia, III, V and VI abolished the genetic function of the CSB protein in survival, RNA synthesis recovery and apoptosis after UV treatment. Similarly, the same mutants failed to complement the sensitivity toward 4-NQO. Thus, the integrity of these helicase motifs is important for the biological function of the CSB protein. On the contrary, a point mutation in a C-terminal, second, NTB motif of the CSB protein showed full complementation in the ability to repair damage induced by UV light or 4-NQO, suggesting that this motif is not important for the CSB repair function. PMID- 11867211 TI - Regulation of novel members of the Arabidopsis thaliana CCAAT-binding nuclear factor Y subunits. AB - Nuclear factor Y (NF-Y) is a highly conserved trimeric activator that recognizes with high specificity and affinity the widespread CCAAT box promoter element. We previously cloned the genes of 23 NF-Y genes of Arabidopsis thaliana (Gene 264 (2001) 173). Now that the Arabidopsis genome sequencing project is complete, we present the cloning, alignments and expression profiles of the remaining six genes coding for the three NF-Y subunits. Consistent with our previous reports, most of the new members of the three subunits show a unique tissue-specific pattern, while another AtNF-YC9 is rather ubiquitous. PMID- 11867212 TI - Cloning, expression analysis, and structural characterization of seven novel human ADAMTSs, a family of metalloproteinases with disintegrin and thrombospondin 1 domains. AB - ADAMTS (A Disintegrin And Metalloproteinase domain, with ThromboSpondin type-1 modules) is a recently described family of zinc-dependent proteases which play important roles in a variety of normal and pathological conditions, including arthritis and cancer. In this work, we report the identification and cloning of cDNAs encoding seven new human ADAMTSs. These novel enzymes have been called ADAMTS-13, -14, -15, -16, -17, -18, and -19. All of them show a domain organization similar to that of previously characterized family members, consisting of a signal sequence, a propeptide, a metalloproteinase domain, a disintegrin-like domain, a cysteine-rich region, and a variable number of TS-1 repeats. Expression analysis revealed that these ADAMTS genes are mainly expressed in fetal tissues, especially in lung (ADAMTS14, ADAMTS16, ADAMTS17, ADAMTS18, and ADAMTS19), kidney (ADAMTS14, ADAMTS15, and ADAMTS16), and liver (ADAMTS13, ADAMTS15 and ADAMTS18). Reverse transcriptase--polymerase chain reaction analysis also revealed the expression of some of these new ADAMTSs in different human adult tissues, such as prostate (ADAMTS13, ADAMTS17, and ADAMTS18), and brain (ADAMTS13, ADAMTS16, ADAMTS17, and ADAMTS18). High levels of ADAMTSs transcripts were also observed in some tumor biopsies and cells lines, including osteosarcomas (ADAMTS19), melanoma and colon carcinoma cells (ADAMTS13). Chromosomal location analysis indicated that the seven identified ADAMTS genes are dispersed in the human genome mapping to 9q34, 10q21, 11q25, 5p15, 15q24, 16q23, and 5q31, respectively. According to these results, together with a comparative analysis of ADAMTSs in other eukaryotic organisms, we conclude that these enzymes, with at least 18 distinct members encoded within the human genome, represent an example of a widely expanded protease family during metazoan evolution. PMID- 11867213 TI - In vitro selection of high affinity HspR-binding sites within the genome of Helicobacter pylori. AB - The major chaperone genes of Helicobacter pylori are negatively regulated by HspR, a homologue of the repressor of the dnaK operon of Streptomyces coelicolor. Using an in vitro selection and amplification approach we identified two new chromosomal binding sites of the HspR protein. Both binding sites were characterized by footprinting analysis with purified HspR protein. Intriguingly, these HspR binding sites are located at the 3prime prime or minute ends of two genes coding for predicted proteins with functions unrelated to those of chaperones. This suggests that H. pylori HspR may regulate the expression of genes encoding proteins with diverse functions. Nucleotide sequence alignment of HspR-binding sites highlights conserved nucleotides extending outside the previously proposed consensus binding sequence with structural features predicting geometry of HspR binding as an oligomer. PMID- 11867214 TI - An integrated genetic, radiation hybrid, physical and transcription map of a region of distal mouse chromosome 12, including an imprinted locus and the 'Legs at odd angles' (Loa) mutation. AB - A variety of loci with interesting patterns of regulation such as imprinted expression, and critical functions such as involvement in tumour necrosis factor pathways, map to a distal portion of mouse chromosome 12. This region also contains disease related loci including the 'Legs at odd angles' mutation (Loa) that we are pursuing in a positional cloning project. To further define the region and prepare for comparative sequencing projects, we have produced genetic, radiation hybrid, physical and transcript maps of the region, with probes providing anchors between the maps. We show a summary of 95 markers and 91 genomic clones that has enabled us to identify 18 transcripts including new genes and candidates for Loa which will help in future studies of gene context and regulation. PMID- 11867215 TI - Identification and characterization of a novel member of olfactomedin-related protein family, hGC-1, expressed during myeloid lineage development. AB - We have cloned a novel hematopoietic granulocyte colony-stimulating factor (G CSF)-induced olfactomedin-related glycoprotein, termed hGC-1 (human G-CSF stimulated clone-1). mRNA differential display was used in conjunction with a modified two-phase liquid culture system. Cultures were enriched for early precursors of erythroid, myeloid, and megakaryocytic lineages, which were isolated after induction with erythropoietin, G-CSF, and thrombopoietin, respectively. RNA from the enriched cells was subjected to differential display analysis to identify lineage-specific expressed genes. One clone specifically induced by G-CSF, hGC-1, was characterized. The 2861 bp cDNA clone of hGC-1 contained an open reading frame of 1530 nucleotides, translating into a protein of 510 amino acids with a signal peptide and six N-linked glycosylation motifs. The protein sequence of hGC-1 showed it to be a glycoprotein of the olfactomedin family, which includes olfactomedin, TIGR, Noelin-2 and latrophilin-1. Olfactomedin-like genes show characteristic tissue-restricted patterns of expression; the specific tissues expressing these genes differ among the family members. hGC-1 was strongly expressed in the prostate, small intestine, and colon, moderately expressed in the bone marrow and stomach, and not detectable in other tissues. In vitro translation and ex vivo expression showed hGC-1 to be an N-linked glycoprotein. The hGC-1 gene locus mapped to chromosome 13q14.3. Together, our findings indicate that hGC-1 is primarily expressed as an extracellular olfactomedin-related glycoprotein during normal myeloid-specific lineage differentiation, suggesting the possibility of a matrix-related function for hGC-1 in differentiation. PMID- 11867216 TI - Characterization of omp200, a porin gene complex from Bacteroides fragilis: omp121 and omp71, gene sequence, deduced amino acid sequences and predictions of porin structure. AB - The high MW porin protein complex (Omp200, composed of Omp121 and Omp71) from Bacteroides fragilis ATCC 25285 was purified and tryptic peptide sequences were used to design degenerate oligonucleotide primers which were then used as a first step in amplification, identification and sequencing of the omp121 gene (GenBank Accession Number AF357210). Sequence analysis revealed an open reading frame of 3378 bases. The deduced amino acid sequence (which contained the experimentally determined peptide sequences) has 1125 or 1116 amino acids (depending on which start codon is used); the mature protein consists of 1096 amino acids, has a predicted MW of 121.4 and a theoretical pI of 6.32. It is preceded by a 29 or 18 amino acid signal peptide which includes a typical hydrophobic region near the N terminus (VLVLVL). Hydropathy plots of the deduced amino acid sequence of B. fragilis Omp121 display striking similarity with those of Escherichia coli OmpC (a 16-stranded porin) and FepA (a 22-stranded ligand-gated transport protein). Three-dimensional modeling of B. fragilis Omp121 (based on 1D and 3D sequence profiles, coupled with secondary structure and solvation potential information) indicated that the closest homologues in terms in fold conservation were the E. coli 16-stranded porins (e.g. OsmA) and 22-stranded ligand gated transport proteins (e.g. FepA). The omp71 gene sequence was identified using the tryptic peptides to search the published Bacteroides genome data base. We found that omp71 is located immediately downstream of omp121 and confirmed this with PCR analysis. Omp71 has no known homologues but does share some characteristics with the Porphyromonas RagB antigen. PMID- 11867217 TI - A carboxylesterase from the hyperthermophilic archaeon Sulfolobus solfataricus: cloning of the gene, characterization of the protein. AB - A genomic library of the hyperthermophilic archaeon Sulfolobus solfataricus strain MT4 was constructed in Escherichia coli using a cloning vector not designed for heterologous gene expression. One positive clone exhibiting acquired thermophilic acetylesterase activity was directly detected by an in situ plate assay using a colony staining procedure with the chromogenic substrate beta naphthyl acetate. The plasmid isolated from the clone contained a 3.3 kb genomic fragment from S. solfataricus and a full-length esterase coding sequence could be identified. Expression of the active thermostable esterase in E. coli was independent of isopropyl-beta-D-thiogalactopyranoside and of the kind of vector, suggesting that the archaeal esterase gene was controlled by fortuitous bacterial like sequences present in its own 5' flanking region, not by the bacterial lac promoter or other serendipitous vector-located sequences. The protein, partially purified by thermoprecipitation of the host proteins at high temperature and gel exclusion chromatography, showed a homo-tetrameric structure with a subunit of molecular mass of 32 kDa which was in perfect agreement with that deduced from the cloned gene. The same protein was revealed in S. solfataricus cell extracts, thus demonstrating its functional occurrence in vivo under the cell culture conditions tested. The recombinant enzyme exhibited high thermal activity and thermostability with optimal activity between pH 6.5 and 7.0. The hydrolysis of p nitrophenyl esters of fatty acids (from C(2) to C(8)) allowed the enzyme to be classified as a short length acyl esterase. PMID- 11867218 TI - Murine Tbx2 contains domains that activate and repress gene transcription. AB - T-box (Tbx) genes represent a phylogenetically conserved family of transcription factors that play important roles during embryonic development. Tbx family members have been shown to either activate or inhibit gene expression. However, little is known about the domains within Tbx proteins responsible for mediating gene transcription. While Tbx2 is known to repress gene expression, the domain(s) within Tbx2 remains poorly defined. Deletion of the carboxy-terminus of Tbx2, which contains a domain that is highly conserved with Tbx3 and ET, which has been demonstrated to contain a repression domain, only minimally diminishes the ability of Tbx2 to repress gene expression. However, in combination with the carboxy-terminal truncation, deletion of the amino acids located amino-terminal to the T-box abolished the ability of Tbx2 to repress gene expression. Both of these domains were capable of repressing gene expression when linked to the GAL4 DNA binding domain. In contrast to these two repression domains, the T-box was capable of weakly activating gene expression depending on the promoter context. Deletion analysis of the T-box suggests that this activation domain is located in the amino-terminal end of the T-box. These results reveal a novel transcription repression domain, confirm the presence of a previously implicated domain, and suggest a novel role for the T-box. Taken together, these results provide the basis for understanding the molecular mechanism whereby Tbx2 regulates gene expression and subsequently controls embryonic development. PMID- 11867219 TI - Diversity of Streptococcus mutans bacteriocins as confirmed by DNA analysis using specific molecular probes. AB - Mutacin-producing strains have been classified into 24 groups (designated by letters A to X) by similarity in activity spectra and cross-immunity. Similarity in primary structure among these groups can be revealed using DNA hybridization. The amino acid sequences of four mutacins (B-Ny266, 1140/mutacin III and mutacin II) were used to design two DNA probes in order to detect similar genes among groups of Streptococcus mutans strains demonstrating inhibitory activity. In addition to the appropriate parent strain, each probe hybridized with the total DNA from only two out of the 24 mutacin group type strains. Thus, the remaining 18 groups of strains produce mutacins that differ from the mutacins sequenced to date. In order to explore the similarity between genes coding for mutacins B Ny266 and JH1140, the group B specific probe was utilized to detect a DNA fragment of 1.9 kb in the genome of S. mutans strain Ny266. The sequence of the cloned fragment codes for three open reading frames (lanA, lanA' and lanB) similar to those of strains JH1140 and UA787. The gene lanA' is strongly similar to the structural gene lanA (67%), but only one RNA transcript of about 300 bases was detected by Northern hybridization using the lanA-lanA' probe. Transcription of lanA alone was verified by RT-PCR. PMID- 11867220 TI - Functional analysis of the promoter of human sterol 27-hydroxylase gene in HepG2 cells. AB - Human sterol 27-hydroxylase catalyses the first step in the alternative pathway of bile acids biosynthesis in hepatocytes. However the gene encoding this enzyme (CYP27 gene) is expressed in every tissue and some evidence suggests that this enzyme plays a role in cholesterol homeostasis. Although modulation of CYP27 expression has been reported, the mechanisms underlying the regulation of this gene in human tissues is still poorly understood. To elucidate the mechanism governing CYP27 expression we cloned a 4.3 kb fragment of the 5' flanking region of the human CYP27 gene and constructed deletion mutants which were transfected into HepG2 cells. Functional assays showed that the -217/-10 nucleotide region from the translation start site (minimal promoter), devoid of TATA and CAAT boxes, contains all the elements for basal transcription. Foot-printing analysis of minimal promoter showed four protected regions (A-D). Regions A, B and D each contain one Sp1 binding site, and region C contains a HNF4 site. Electrophoretic mobility shift assays demonstrated that Sp1, Sp3 and HNF4 transcription factors bind these sites. Mutagenesis of any of these sites resulted in the loss of promoter activity. Co-transfection of the minimal promoter with Sp1 and Sp3 expression vectors transactivated CYP27 gene promoter in Drosophila SL2 cells, which lack endogenous Sp proteins. Transactivation of the minimal promoter was also observed in HeLa cells co-transfected with HNF4 expression vector. Therefore, Sp1, Sp3 and HNF4 co-operate in the expression of the human CYP27 gene in HepG2 cells. PMID- 11867221 TI - Characterization of a novel gene up-regulated during anoxia exposure in the marine snail, Littorina littorea. AB - Gene expression was investigated during anoxia exposure in the marine snail, Littorina littorea. Differential screening of a cDNA library made from hepatopancreas of anoxic L. littorea yielded a 525 bp clone coding for the novel gene kvn. The deduced amino acid sequence of the KVN protein contained 99 amino acid residues with a predicted molecular weight of 12 kDa and showed an N terminal secretory signal. Analysis of hepatopancreas samples over a time course of anoxia exposure showed a maximum increase in transcript levels of 5.8-fold after 48 h relative to normoxic animals, with a subsequent decrease in transcript levels during normoxic recovery. Nuclear run-off assays confirmed the observed transcriptional up-regulation of kvn during anoxia. Organ culture experiments were performed to determine a possible pathway of up-regulation of kvn, with data indicating a putative role for cGMP in signal transduction. Profiles of ribosome distribution in polysomes versus monosomes revealed a reduction in the polysome peak during anoxia and a shift in the position of kvn transcripts to association with the lower density polysome/higher density monosome region. The data suggest that the kvn transcript is both transcribed and translated during anoxia, indicating a possible significant role for the KVN protein in the survival of anoxia by L. littorea. PMID- 11867222 TI - Structure of the rabbit alphas1- and beta-casein gene cluster, assignment to chromosome 15 and expression of the alphas1-casein gene in HC11 cells. AB - Several casein (CSN) genes (CSN1, 2, 10 and alphas2-CSN) have been described and shown to be clustered in mouse, man and cattle. These genes are expressed simultaneously in the mammary gland during lactation, but they are silent in most mammary cell lines, even in the presence of lactogenic hormones. However, it has been shown that the CSN2 gene, and this gene only, can be induced in certain mammary cell lines, such as HC11. In the present paper, we describe three overlapping bacterial artificial chromosome (BAC) clones which harbor both the rabbit CSN1 and CSN2 genes. These two genes are in a convergent orientation, separated by an intergenic region of 15 kb. DNA from one of the CSN/BAC clones was used as a probe for in situ hybridization to show that the CSN1 and CSN2 gene cluster is located on chromosome 15 band q23 and not on chromosome 12 as had been previously reported. Each of the three CSN/BAC DNAs was transfected into HC11 cells. In the presence of lactogenic hormones, the rabbit CSN1 gene was clearly expressed from all three CSN/BAC DNAs, whereas the rabbit CSN2 gene, which at the most possesses a 1 kb upstream region in one of the CSN/BAC DNAs, was not expressed at detectable levels on Northern blots. The transfected HC11 cells now express both rabbit CSN1 and mouse CSN2 genes. These transfected cells will be used as a model to study the role of CSN1 in milk protein secretion. PMID- 11867223 TI - An ADAM family member with expression in thymic epithelial cells and related tissues. AB - We have analyzed the tissue-specific expression, mRNA isoforms, and genomic structure of murine ADAM28, an ADAM family member recently discovered in human and mouse. While human ADAM28 is expressed in lymphocytes (J. Biol. Chem. 274 (1999) 29251), we observe expression of murine ADAM28 in thymic epithelial cells and developmentally related tissues including the trachea, thyroid, stomach, and lung, but not in lymphocytes. The expression patterns in adult and day 15.5 embryos are similar. We have detected multiple mRNA isoforms varying in the cytoplasmic domain coding sequence and 3prime prime or minute untranslated region due to alternative polyadenylation and splicing events that occur in the final four exons and three introns. The entire ADAM28 gene spans 55 kb and contains 23 exons. The protein sequence contains all conserved residues required for metalloprotease activity, indicative of a role in ectodomain shedding and extracellular matrix modeling. Given its unique expression pattern and potential functions, murine ADAM28 may play a role in organogenesis and organ-specific functions such as thymic T cell development. PMID- 11867224 TI - Cloning and characterization of two subunits of Arabidopsis thaliana nuclear cap binding complex. AB - In this report we characterize two Arabidopsis thaliana proteins, named AtCBP20 and AtCBP80, that are homologues of human subunits of a nuclear cap-binding protein complex (CBC). AtCBP20 has a calculated molecular mass of 29.9 kDa, and AtCBP80 is a 96.5 kDa protein. AtCBP20 exhibits 68% identity and 82% similarity to human CBP20. Like its human homologue, AtCBP20 contains a canonical RNA binding domain (RBD) with single RNP2 and RNP1 motifs. In addition to the N terminal part, which is similar to the human protein, AtCBP20 has a long C terminus rich in arginine, glycine and aspartate residues. The second subunit of the Arabidopsis cap-binding complex, AtCBP80, shows 28% identity and 50% similarity to its homologue from HeLa cells. The protein contains a MIF4G domain at its N-terminus, the feature characteristic to all analyzed CBP80s. This domain, described also in eIF4G and NMD2 proteins, is thought to be involved in protein-protein and also in protein--RNA interactions. Both proteins AtCBP20 and AtCBP80 are encoded by single-copy genes in the A. thaliana genome. The AtCBP20 gene is located on chromosome V, and the AtCBP80 gene is encoded by chromosome II. Among introns identified in the AtCBP20 gene, we discovered an U12 type intervening sequence (an AT-AC intron). This intron is spliced out very efficiently in plants, but when isolated and tested for splicing in tobacco protoplasts, the efficiency of the U12 intron excision was low. Splicing efficiency of the U12 intron is improved by the addition of exon and intron sequences upstream or downstream of the U12 intron. AtCBP20 and AtCBP80 are constitutively expressed in all examined organs of A. thaliana, including roots, stems, leaves and flowers. Interestingly, the steady-state level of both transcripts seem to be very similar in all tissues analyzed. PMID- 11867226 TI - Expression from cell type-specific enhancer-modified retroviral vectors after transduction: influence of marker gene stability. AB - The enhanced green fluorescent protein (EGFP) is increasingly used as a reporter gene in viral vectors for a number of applications. To establish a system to study the activity of cis-acting cellular regulatory sequences, we deleted the viral enhancer in EGFP-carrying retroviral vectors and replaced it with cell type specific elements. In this study, we use this system to demonstrate the activity of the human CD2 lymphoid-specific and the Tie2 endothelial cell type-specific enhancers in cell lines and in primary cells transduced by retroviral vectors. Furthermore, we compare findings obtained with EGFP as the reporter gene to those obtained replacing EGFP with d2EGFP, an unstable variant of EGFP characterized by a much shorter half-life compared to EGFP, and by reduced accumulation in the cells. d2EGFP-carrying vectors were generated at titers which were not different from those generated by the corresponding vectors carrying EGFP. Moreover, the activity of a Moloney murine leukemia virus enhancer could be readily detected following transduction of target cells with either EGFP- or d2EGFP-carrying vectors. However, the activity of the relatively weak CD2 and Tie2 enhancers was exclusively detected using EGFP as the reporter gene. These findings indicate that enhancer replacement is a feasible and promising approach to address the function of cell type-specific regulatory elements in retroviral vectors carrying the EGFP gene. PMID- 11867225 TI - Functional and binding studies of HS3.2 of the beta-globin locus control region. AB - The distal locus control region (LCR) is required for high-level expression of the complex of genes (HBBC) encoding the beta-like globins of mammals in erythroid cells. Several major DNase hypersensitive sites (HSs 1-5) mark the LCR. Sequence conservation and direct experimental evidence have implicated sequences within and between the HS cores in function of the LCR. In this report we confirm the mapping of a minor HS between HS3 and HS4, called HS3.2, and show that sequences including it increase the number of random integration sites at which a drug resistance gene is expressed. We also show that nuclear proteins including GATA1 and Oct1 bind specifically to sequences within HS3.2. However, the protein Pbx1, whose binding site is the best match to one highly conserved sequence, does not bind strongly. GATA1 and Oct1 also bind in the HS cores of the LCR and to promoters in HBBC. Their binding to this minor HS suggests that they may be used in assembly of a large complex containing multiple regulatory sequences. PMID- 11867227 TI - hUNC93B1: a novel human gene representing a new gene family and encoding an unc 93-like protein. AB - We have identified a novel human gene UNC93B1 encoding a protein related to unc 93 of Caenorhabditis elegans. The combined sequence derived from several cDNA clones is 2282 bp and comparison with genomic sequence shows that the gene contains 11 exons. The longest open reading frame encodes a deduced sequence of 597 amino acids. Homology analysis shows that the hUNC93B1 gene is highly conserved and related to sequences in Arabidopsis thaliana, C. elegans, Drosophila melanogaster, chicken and mouse. Structural analysis of the deduced amino acid sequence of hUNC93B1 points to possible existence of multiple membrane spanning domains. hUNC93B1 protein also displays some similarities to the bacterial ABC-2 type transporter signature and to ion transporters of Deinococcus radiodurans and Helicobacter pylori. As revealed by Northern analysis, the level of expression varies significantly between tissues, with the highest level detected in the heart. The gene was mapped to chromosomal band 11q13 by fluorescence in situ hybridization. We suggest that this gene is a member of a novel hUNC93B-related gene family. PMID- 11867228 TI - Genomic organization of profilin-III and evidence for a transcript expressed exclusively in testis. AB - Profilins are small, widely expressed actin binding proteins, thought to be key regulators of actin dynamics in living cells. So far, three profilin-genes have been described: profilin-I (PFN1), profilin-II (PFN2) with two splice variants and the recently identified profilin-III (PFN3). Here we describe the genomic organization of the genes encoding human and mouse profilin-III. Both are single exon genes and lie in close vicinity to the renal sodium-phosphate transport gene 2 (SLC34A1, NPT2) which is highly expressed in kidney. Northern hybridization to rat tissues has previously demonstrated expression of an approximately 4.5 kb long profilin-III mRNA transcript in kidney and a mRNA transcript of approximately 1 kb in length in testis. Here we show that mouse profilin-III expression is restricted to testis and that the 4.2 kb profilin-III mRNA in kidney is the result of a slc34a1 transcript which includes the antisense profilin-III open reading frame in its 3'-untranslated region. Finally, we demonstrate by in situ hybridization that profilin-III mRNA is localized to cells in the late stage of spermatogenesis. PMID- 11867229 TI - Effects of signaling molecules, protein phosphatase inhibitors and blast pathogen (Magnaporthe grisea) on the mRNA level of a rice (Oryza sativa L.) phospholipid hydroperoxide glutathione peroxidase (OsPHGPX) gene in seedling leaves. AB - With a specific focus on rice self-defense response(s), the effects of global signaling molecules, jasmonic acid (JA), salicylic acid (SA), abscisic acid (ABA), and ethylene (using the ethylene generator, ethephon), and protein phosphatase (PP) inhibitors, cantharidin and endothall on expression of a rice phospholipid hydroperoxide glutathione peroxidase (OsPHGPX) gene in rice seedling leaves were investigated. We provide first evidence for a potent up-regulation of the OsPHGPX mRNA accumulation by these signaling molecules and PP inhibitors that strongly suggest its potential role in defense/stress. The OsPHGPX gene also showed a weak constitutive expression and responsiveness to cut. These inductions were influenced by light signal(s), and did not show a requirement for de novo synthesized protein factor(s). A potential interaction amongst these signaling molecules, especially JA, SA, ABA and kinetin, in modulating the OsPHGPX expression was found. The blast pathogen, Magnaporthe grisea also elicited the accumulation of OsPHGPX mRNA in leaves. This is a first systematic report in rice (and in plants) demonstrating the inducible nature (and expression) of the OsPHGPX gene by a variety of defense/stress-related stimuli, and modulation by the PPs of the kinase-signaling cascade(s). PMID- 11867230 TI - Cell line-specific translation of two laminin 5 beta3 chain isoforms. AB - In sequencing the beta3 chain of laminin 5 mRNA from LNCaP cells, we observed three different human cDNA clones (XM_001716, NM_000228 and L25541) in the GenBank that identified different sequences in the untranslated regions (UTR). XM_001716 and NM_000228 are almost identical cDNA clones with approximately 99% homology. However, they are quite different from L25541 in both the 5' UTR and the 3' UTR. Development of a PCR assay to specifically detect two of these different forms of the message led to the observation that they were differentially expressed in various cell lines. The message designated B3A (NM_000228, and XM_001716) was absent in LNCaP and MCF7 and greatly reduced in PC3-N, but was present in eight other epithelial cell lines. B3B (L25541) was present in all cell lines studied. The cell lines that failed to express the B3A form also failed to express the protein based on both immunoblotting and immunohistochemical analysis. It appears from this data that there are two isoforms of the beta3 mRNA, and that the 5' UTRs of the mRNAs play an important role in regulating translation of the beta3 protein. Since laminin 5 is lost in prostate carcinoma, the mechanism of control that results in the translation of the two forms of message may be important in tumorigenesis. PMID- 11867231 TI - Identification and characterization of ameloblastin gene in a reptile. AB - Ameloblastin (AMBN) is one of the enamel sheath proteins which presumably has a role in determining the prismatic structure of growing enamel crystals. There may therefore be a relationship between the molecular evolution of the AMBN gene and the development of enamel prismatic structures. To investigate whether such a relationship exists, it was necessary to identify the homologues of the AMBN gene in a reptile whose teeth lack an enamel prismatic structure. To this end, several clones containing AMBN cDNA were isolated from caiman jaws using the reverse transcription-polymerase chain reaction (RT-PCR) method. Sequence analysis of the AMBN cDNA revealed an open reading frame of 1221 bp encoding a 407-amino-acid protein. Translation of the caiman cDNA starts at the methionine corresponding to the second of two putative start codons conserved in mammalian AMBN genes. The N terminal part of the caiman AMBN shows high amino acid sequence similarities to human, pig, cattle, rat and mouse AMBN sequences, as well as several other features that have been conserved throughout the evolution of reptiles and mammals. Unexpectedly, the nucleotide sequences of the 3' untranslated region (UTR) are also conserved, not only within mammalian genes but also between reptilian and mammalian genes. The caiman AMBN gene is a single-copy gene, transcribed only in the jaws, presumably in teeth. PMID- 11867233 TI - DNA-PKcs mutations in dogs and horses: allele frequency and association with neoplasia. AB - Previously, spontaneous genetic immunodeficiencies in mice, Arabian foals, and recently in Jack Russell terriers have been ascribed to defects in DNA-PKcs (catalytic subunit of the DNA dependent protein kinase) expression. In severe combined immunodeficiency (SCID) foals, a 5 bp deletion at codon 9480 results in a frameshift and a 967 amino acid deletion from the C terminus (including the entire PI3 kinase domain) and an unstable mutant protein. In SCID mice, a single base pair mutation results in a premature stop codon and deletion of 83 amino acids; as in SCID foals, the mutant protein is unstable. Here, we define the mutation within the canine DNA-PKcs gene that results in SCID. In this case, a point mutation results in a stop codon at nucleotide 10,828 and premature termination at a position 517 amino acids before the normal C terminus resulting in a functionally null allele. Thus, this is the third documentation of a spontaneous germline mutation in the C terminus of DNA-PKcs. Emerging data implicate DNA repair factors as potential tumor suppressors. Here, we have ascertained the carrier frequency of the defective DNA-PKcs genes in Arabian horses and in Jack Russell terriers. Our data indicate (in good agreement with a previous report) that the carrier frequency of the equine SCID allele is approximately 8%; in contrast, the carrier frequency of the canine SCID allele is less than 1.1%. We also assessed the frequency of the equine SCID allele in a series of 295 tumors from Arabian horses. We find a statistically significant correlation between the development of a virally induced tumor (sarcoid) and heterozygosity for the equine SCID allele. These data provide further support for an emerging consensus: that DNA-PK may normally act as a tumor suppressor through its caretaker role in maintaining chromosomal stability. PMID- 11867232 TI - Sequencing of the rat beta-catenin gene (Ctnnb1) and mutational analysis of liver tumors induced by 2-amino-3-methylimidazo[4,5-f]quinoline. AB - beta-Catenin, a protein that functions in cadherin-mediated cell-cell adhesion as well as in signal transduction, has received increasing attention in recent years due to its role as an oncogene in various human cancers. The primary sequence of the human beta-catenin gene (CTNNB1) has been known for some time, but that of the rat beta-catenin gene (Ctnnb1) has not heretofore been studied in detail. We report here the primary structure of Ctnnb1 using PCR-based methods and direct sequencing. The size of the complete Ctnnb1 gene was determined to be 9082 bp. We found the rat Ctnnb1 gene to contain 14 exons, ranging in size from 61 to 356 bp, and 13 introns ranging in size from 76 to 2524 bp. The transcription start site appears to be 157 bp upstream of the ATG codon located in exon 1. The resulting transcript is 2650 nucleotides long (encoding a protein of 781 amino acids). We found the 5' UTR to consist of 157 nucleotides and the 3' UTR to be 147 nucleotides long. The region coding for the glycogen synthase kinase-3beta domain of beta-catenin is located in exon 2 of rat Ctnnb1, in contrast to human CTNNB1 in which it is found in exon 3. Based on the newly acquired knowledge of the primary sequence, more than a dozen 2-amino-3-methylimidazo[4,5-f]quinoline (IQ) induced rat liver tumors were screened for the presence or absence of mutations in all 14 exons of rat Ctnnb1. Surprisingly, no mutations were found. The results are discussed in the context of the organ-specificity of IQ-induced mutations in beta-catenin, being highly prevalent in colon tumors, but much less common in liver tumors. PMID- 11867234 TI - Cloning the AFURS1 gene which is up-regulated in senescent human parenchymal kidney cells. AB - To study the changes in gene expression in senescent cells we applied the suppression subtractive hybridization of two cDNA pools isolated from human parenchymal kidney cells in the phase of exponential growth and cellular senescence in vitro. In addition to several genes known to be associated with cellular senescence, we identified a new gene, which is overexpressed in senescent kidney parenchymal cells. The full-length cDNA consists of 5226 nucleotides with an open reading frame (ORF) encoding 701 amino acids (Accession number: AJ306929). The gene product has a predicted molecular mass of 77.31 kDa. The ORF of the new gene shows significant homology to P-type ATPase family gene products and therefore was called AFURS1 (ATPase family homolog up-regulated in senescence cells). The consensus sequence phosphorylation site (DKTGTLT) is highly conserved. According to the GenBank database AFURS1 is mapped to the sequence segment NT_005535.3 at chromosomal region 3q26.32 and has 18 exons. The AFURS1 gene might have a role in cellular aging and tumor suppression as well. PMID- 11867235 TI - Cloning and sequence analysis of a novel metalloprotease gene from Vibrio parahaemolyticus 04. AB - The metalloprotease gene (vppC) from Vibrio parahaemolyticus 04 has been cloned and sequenced. The vppC gene contains an open reading frame of 2442 nucleotides encoding a polypeptide of 814 amino acids with a calculated molecular mass of 89,833 Da. The predicted amino acid sequence of VppC containing a zinc metalloprotease HEXXH consensus motif displays extensive homology to the collagenase from Vibrio alginolyticus. The activity of the recombinant protease produced in Escherichia coli was examined by gelatin zymography and proteolytic activity assays. The substrate specificity study showed that the type I collagen and synthetic collagenase substrate carbobenzoxy-glycyl-L-prolyl-glycyl-glycyl-L prolyl-L-alanine were the best substrates, indicating that the cloned metalloprotease is indeed a collagenase. Multiple alignment analysis of the amino acid sequences and the enzymatic properties such as molecular mass and substrate specificity revealed three distinct classes of Vibrio metalloproteases. The identification of a new metalloprotease gene expands the role of Vibrio metalloproteases as a virulence factor for host infection. PMID- 11867236 TI - Expression of novel lipocalin-like milk protein gene is developmentally-regulated during lactation in the tammar wallaby, Macropus eugenii. AB - We have identified a novel whey protein (late lactation protein B; LLPB) that is first secreted in the milk of the tammar wallaby around day 200 of lactation. The LLPB cDNA clone of 843 base pairs encodes a mature protein of 156 amino acids. LLPB shares 65 and 48% nucleotide and deduced amino acid identity, respectively, with a previously identified late lactation protein A (LLPA). Both these proteins share significant amino acid sequence homology with the lipocalin protein family. Expression of the LLPB gene is induced between days 200 and 240 of lactation, in contrast to expression of the LLPA gene, which is induced at around 145 days of lactation. Maximal expression of both genes in mammary explants from tammars at 213 days of lactation required a combination of prolactin, insulin and hydrocortisone. Transcripts of LLPA, LLPB and beta-lactoglobulin (TBLG) were localized to the same cells by in situ hybridization. A substantial level of alveolar maturation is required for expression of the LLP genes, unlike TBLG, which is expressed in immature alveoli. We hypothesize that the temporal expression of the LLPB and LLPA genes may be regulated both by endocrine stimuli and factors intrinsic to the mammary gland. PMID- 11867237 TI - Molecular cloning of XTP, a tau-like microtubule-associated protein from Xenopus laevis tadpoles. AB - The microtubules of the mammalian nervous system are stabilised by several microtubule-associated proteins (MAPs), including the tau and MAP-2 protein families. The most prominent feature of mammalian tau and MAP-2 proteins is a common and highly homologous microtubule-binding region consisting of three or four imperfect tandem repeats. In this paper we report the cloning and characterisation of a Xenopus laevis tau-like protein (XTP) from tadpole tails. This protein encompasses two isoforms of 673 or 644 amino acids with four tandem repeats that are highly homologous to mammalian tau repeats. Both isoforms share a common amino terminal half, whereas the carboxyl terminus downstream of the repeat region is unique for each isoform. Northern blot analysis revealed that both isoforms are preferentially expressed in the tail of X. laevis tadpoles, whereas a shorter version of XTP is expressed in the head. Recombinant proteins of both XTP isoforms were able to bind microtubules. The longest isoform, however, was more effective at promoting tubulin polymerisation, indicating that sequences downstream of the repeat region affect the microtubule assembling capacity. These results demonstrate that tau-like proteins are found in non mammalian vertebrate species, where they may support the stability of microtubules. PMID- 11867238 TI - The treatment of HIV/AIDS was revolutionized in the mid 1990's with the advent of highly active antiretroviral therapy (HAART). PMID- 11867239 TI - Patient adherence to HIV medication regimens: a review of published and abstract reports. AB - A literature search was conducted to collect published articles reporting correlates of HIV medication adherence or interventions designed to increase HIV medication adherence. Proceedings from seven HIV/AIDS-related conferences were searched for relevant abstracts. We found 18 descriptive studies in published articles and 57 in conference proceedings producing over 200 separate variables falling into four broad areas: (1) factors related to treatment regimen; (2) social and psychological factors; (3) institutional resources; and (4) personal attributes. More complex regimens were related to decreased adherence, but were often successfully mitigated by regimen aids. Social and psychological factors reflecting emotional adjustment to HIV/AIDS and provider support were related to adherence. Access to institutional resources was associated with better adherence. Personal attributes showed a mixed relationship; gender was not consistently related to adherence, but younger age, minority status, and a history of substance abuse were often related to non-adherence. The intervention search yielded 16 interventions employing a wide range of behavioral, cognitive and affective strategies. Evidence of effectiveness was weak. We conclude the abstracts are a useful source of information as part of a systematic review, particularly when available published literature is limited, if results and study characteristics are reported in an adequate and standard manner. PMID- 11867240 TI - Internet access and Internet use for health information among people living with HIV-AIDS. AB - Widespread access to the Internet has the potential to improve the health care and quality of life of people with chronic illnesses, including people living with HIV-AIDS. However, the Internet is not equally accessible to all persons. We surveyed 96 men and 51 women living with HIV-AIDS regarding their experiences using the Internet. Results showed that persons with 12 or fewer years of education were significantly less likely to have used the Internet and were less likely to have been instructed in Internet use. A broad range of health-related Internet activities was reported including searching for health, AIDS-specific information, and using the Internet to communicate with providers. Among current Internet users, individuals who had an Internet connection in their home reported significantly more experiences using the Internet, including Internet use for interpersonal communication and search functions. A digital divide therefore exists among people living with HIV-AIDS, and the benefits of the Internet appear better achieved with home access. PMID- 11867241 TI - Development of a tuberculosis education booklet for Latino immigrant patients. AB - The incidence and prevalence of tuberculosis are far more common among third world populations immigrating to the United States than among US-born citizens. Immigrants' failure to comply with an anti-tuberculosis treatment can impede completion of care and further confound this public health problem. Barriers to patient--provider communication can negatively influence adherence to a medical regimen. Patients who are unable to comprehend medical advice and do not see it as personally salient are less likely to follow their provider's medical advice. In this paper, the authors focus on efforts to develop a patient education tool targeting Spanish-speaking Latino immigrant patients to facilitate communication with tuberculosis clinicians. A description of the multi-stage developmental processes is presented including conducting a needs assessment, development of visual and written messages, review/critique by tuberculosis experts, field testing, revisions, and distribution. Formative evaluation and field testing indicates promise for improving communication using this tool. PMID- 11867242 TI - Changes in eating and weight control habits after myocardial infarction. AB - The focus of this study was to describe changes the patients make in eating and weight control habits 3--4 months after diagnosis of myocardial infarction, and to identify difficulties associated with the reported changes 1 month later. Subjects were 50 male patients (mean age 48 years, mean body mass index 27.9kg/m(2)) participating in an in-patient cardiac rehabilitation programme. The changes were assessed with a questionnaire (46 items) at the beginning of the rehabilitation programme and 4 days food diaries were used to assess the diet at home. Difficulties were assessed with a questionnaire (34 items) 1 month later. Nearly all patients (96%) had changed their habits after myocardial infarction. The majority of the patients (76%) reported that they had consumed foods containing less fat and many of them anticipated difficulties to eat healthily at workplace. More attention could be paid to self-change without the assistance of formal treatment among patients with cardiac disease. PMID- 11867243 TI - Timing of patient satisfaction assessment: effect on questionnaire acceptability, completeness of data, reliability and variability of scores. AB - The present study compared the performance of a multidimensional patient satisfaction questionnaire according to the timing of questionnaire administration. Comparisons were made in terms of: (a) the completeness and representativeness of the data set (number of missing questionnaires, number missing item responses, respondents' representativeness to the target population); (b) the questionnaire acceptability to respondents (time and difficulty to complete); (c) the questionnaire reliability; and (d) variability of scores. One hundred and ten consecutive breast cancer patients hospitalised for surgery were randomised between being sent the comprehensive assessment of satisfaction with care (CASC) at 2 weeks (T(2W)) or at 3 months (T(3M)) after hospital discharge. The time to complete the CASC was shorter at T(3M) than at T(2W) and the mean percentage of item omission was lower at T(3M) (1.68) than at T(2W) (3.82). However, the response rate was much higher at T(2W) (87%) than at T(3M) (66%), making item omission non-significant. At both times of questionnaire administration samples were equally biased towards patients having undergone a less invasive surgery. Moreover, the multi-item scales of the CASC demonstrated adequate internal consistency coefficients, except the general satisfaction scale at T(3M), and fairly symmetrical distribution of scores. Response rate should be considered in priority. This criteria favoured an administration of the CASC shortly after discharge. Besides in a cancer patient population care experience and perception may vary in a 6 weeks time lapse. The timing of assessment needs to be clearly specified in cancer patients satisfaction survey. PMID- 11867244 TI - Correlates of tobacco use among smokers and recent quitters diagnosed with cancer. AB - Smoking after a cancer diagnosis shortens survival time, increases risk of recurrence and the development of another primary tumor, reduces treatment efficacy, and increases treatment complications. Nevertheless, many patients who smoked prior to their illness continue to smoke after diagnosis and treatment. The development of effective smoking cessation interventions for cancer patients has been slowed by the lack of data concerning psychological correlates of smoking in this population. This study, with 74 cancer patients, showed that smoking and lower readiness to quit was associated with: having relatives at home who smoke, a longer time between diagnosis and assessment, completion of medical treatment, greater nicotine dependence, lower self-efficacy, quitting pros, and risk perceptions, and higher quitting cons, fatalistic beliefs, and emotional distress. Thus, smoking cessation treatments for cancer patients should include pharmacotherapy, relapse prevention, and counseling designed to facilitate self efficacy, quitting pros, and risk awareness and to reduce the quitting cons, fatalism, and distress. PMID- 11867245 TI - A step-wise role playing approach for teaching patient counseling skills to medical students. AB - We describe a step-wise role playing approach to bedside teaching during the clinical training of medical students. The objective of this approach is to teach them the skills which are required to practice patient-centered medicine. "Patient-centered medicine" refers to a style of practice which relates to patients' needs rather than to the doctor's own plan, and which moves from professional control to patient empowerment. Our approach is based on: (a) interviews with real patients, and (b) re-play of doctor--patient encounters in small group teaching sessions using the instructor (a physician) as a simulated patient, while a student assumes the role of the physician. The objective of the simulation is to assess the student's ability to provide health-related information, involve patients in making clinical decisions and plan their management in a manner which suits their preferences and lifestyle. The medical background of the instructor who simulates the patient eliminates barriers in communication and allows these objectives to be easily accomplished. The discussion which follows, attempts to: (a) identify discrepancies between the optimal counseling which was offered to the simulated patient and that offered to the real patient; and (b) show that although inevitable, these discrepancies are not irreducible. We have no formal evaluation of our approach in terms of whether it achieved its objective, produced changes in students' attitudes and bedside manners, or in terms of students' ratings of the teaching approach. However, student participation and occasional verbal feedback have indicated that the teaching intervention may be a valid contribution to the clinical training of medical students and that it may be of use for other clinical instructors. PMID- 11867246 TI - Effects of a novel arginine-vasopressin derivative, NC-1900, on the spatial memory impairment of rats with transient forebrain ischemia. AB - NC-1900, an arginine-vasopressin derivative, has been reported to enhance memory for avoidance behavior. Specifically, NC-1900 ameliorated cycloheximide-induced learning impairments in a passive avoidance test in rats. In the present study, we investigated that effects of NC-1900 on place learning in rats with selective lesions in the CA1 subfield of the hippocampal formation produced by transient forebrain ischemia. NC-1900 was administered daily (1 microg/kg, p.o.) 1 h before the place learning task. A rat was required to alternate between 2 small circular areas located diametrically opposite each other on the circumference of an open field in order to obtain intracranial electrical stimulation reward (the spatial navigation task). Rats with hippocampal lesions showed severe place learning impairments both in task performance (indicated by number of rewards obtained per a session) and in navigation performance (forming efficient trails) over the 30 day test period. Treatment with NC-1900 ameliorated deficits in the place learning exhibited by rats with the same hippocampal lesions, such that their performance reached normal levels. There were no significant differences in the ischemic hippocampal lesions, spontaneous locomotor activity, and stimulation current intensity between the treated and untreated rats. The results demonstrated that NC-1900 reduced place learning impairments produced by hippocampal lesions. PMID- 11867247 TI - Human temporal-lobe response to vocal sounds. AB - Voice is not only the vehicle of speech, it is also an 'auditory face' that conveys a wealth of information on a person's identity and affective state. In contrast to speech perception, little is known about the neural bases of our ability to perceive these various types of paralinguistic vocal information. Using functional magnetic resonance imaging (fMRI), we identified regions along the superior temporal sulcus (STS) that were not only sensitive, but also highly selective to vocal sounds. In the present study, we asked how neural activity in the voice areas was influenced by (i) the presence or not of linguistic information in the vocal input (speech vs. nonspeech) and (ii) frequency scrambling. Speech sounds were found to elicit greater responses than nonspeech vocalizations in most parts of auditory cortex, including primary auditory cortex (A1), on both sides of the brain. In contrast, response attenuation due to frequency scrambling was much more pronounced in anterior STS areas than at the level of A1. Importantly, only right anterior STS regions responded more strongly to nonspeech vocal sounds than to their scrambled version, suggesting that these regions could be specifically involved in paralinguistic aspects of voice perception. PMID- 11867248 TI - The N400 is modulated by unconsciously perceived masked words: further evidence for an automatic spreading activation account of N400 priming effects. AB - It is a matter of debate whether the N400 component of the event-related brain potential (ERP) is sensitive to unconscious automatic priming mechanisms or to strategic mechanisms only. Recent studies demonstrated N400 modulation by masked primes at a short SOA supporting an automatic spreading activation account. However, it cannot be ruled out that strategic mechanisms based upon partial prime identification contributed to the observed priming effects. The present study was set up to substantiate masked N400 priming effects as an index of automatic spreading activation. It was assessed whether partial identification of the masked words due to backward priming could have supported strategic priming to occur. In experiment 1, ERPs were recorded while subjects performed lexical decisions on targets preceded by masked and unmasked primes at an SOA of 67 ms. Masked words, which were not consciously perceived, as well as visible words were shown to modulate the N400 to meaningfully related target words. Experiment 2 required subjects to perform decisions on visual, lexical and semantic features of masked words presented with or without semantically related context words. Subjects performed at chance in all tasks. Furthermore, the results exclude the possibility that backward priming has rendered the masked words partially visible. The present study therefore demonstrates that N400 priming effects can be reliably obtained from unconsciously perceived masked words at a very short SOA and strengthens the notion that the N400 is modulated by automatic spreading activation and not exclusively by strategic semantic processes. PMID- 11867249 TI - Selective and delay adaptation of human saccades. AB - The consistently triggered step back of a target during primary saccades of a human subject induced a gradual change in gain, the ratio of the saccade amplitude to the target eccentricity. After a few hundred trials, subjects were able to foveate the displaced target in a single saccade. Presentation of a displaced target showed that human memory guided saccades have gain adaptation just like the well-established adaptation of visually guided saccades. Examining the transfer of adaptation between the memory guided saccade and two other types of visually guided saccades showed that each saccade transferred a 10-25% adapted gain change to the other saccades. However, any pair of the three saccades acquired different gains by adaptation in the same horizontal direction simultaneously, hence each saccade had adaptive capability independent of the others. Adaptation took place even when the appearance of a displaced target was delayed by 400-600 ms from the end of a primary saccade. These findings have important implications about the adaptation, particularly the location and temporal property of the adaptive mechanism in saccade generation. PMID- 11867250 TI - Corticostriatal circuitry mediates fast-track visual categorization. AB - Previous studies have shown that briefly presented natural scenes containing non animals elicited more negative potentials than images with animals even at 150 ms after stimulus onset (dN150). Cognitive models suggest that both feed-forward and feature weighting processes are involved in the rapid categorization of complex natural scenes. Here we examined the possible neuronal substrates of this model. Patients with Alzheimer's disease (AD) exhibited a delayed dN150, but in their case non-animals evoked more negative potentials similarly to the controls (presence of dN150). In contrast, in patients with Parkinson's disease (PD) animal and non-animal stimuli elicited nearly identical early responses (absence of dN150). The results indicate that when cortico-cortical pathways mediating feed-forward mechanisms are impaired (as in the case of AD), dN150 appears later, while in the case of corticostriatal dysfunctions (as in the case of PD) no differential response is present. This supports the hypothesis that corticostriatal circuits mediate perceptual feature weighting and integration in complex situations requiring categorical judgements. PMID- 11867251 TI - The size of corpus callosum and functional connectivities of cortical regions in finger and shoulder movements. AB - The correlations between the size of corpus callosum and the inter- and intra hemispheric EEG coherence and the spatial EEG synchronization during finger and shoulder movements were analyzed in nine right-handed men. The cross-sectional surface areas of corpus callosum (CC) and of seven callosal regions were measured from the mid-sagittal slice of the anatomical MRI. Movement-related coherence between pairs of EEG electrodes overlying the central and parietal regions of both hemispheres was computed after spatially filtering EEG data by the Laplacian operator method. The spatial EEG synchronization was evaluated using omega complexity, a novel measure which quantifies the number of independent sources of spontaneous EEG oscillations. The amplitude of coherence between the left and right S1/M1 areas after movement onset in the lower alpha band (7.8-9.8 Hz) correlated with the size of the callosal body in both types of movement. The size of the callosal body also correlated with the C3-Cz coherence in the 15.6-19.5 Hz band in finger movement, and in the 15.6-23.5 Hz band in shoulder movements. The size of the rostral, anterior intermediate and posterior intermediate truncus of CC correlated with omega-complexity in both types of movements indicating more foci of synchronized EEG oscillations in subjects with a large callosal truncus. The results suggest that the size of callosal truncus which is known to connect the primary sensorimotor and the supplementary motor areas of both hemispheres contributes to the coupling of EEG oscillations during voluntary finger and shoulder movements. PMID- 11867252 TI - Topography of evoked potentials associated with illusory motion perception as a motion aftereffect. AB - Motion aftereffect (MAE) is a type of motion illusion. After visual focusing on an object moving in one direction, an illusory perception of motion in the opposite direction occurs while the object suddenly stops moving. In this study we explored components and distribution of evoked potentials related to this motion illusion using MAE caused by motion of concentric rings. When a single array of moving rings was placed to straddle right and left visual fields, a significant bilateral increase of a positive component at about 160 ms (P160) was observed in the occipitotemporal region at the time subjects perceived the motion illusion; this increase was most prominent in the right posterior temporal region. Thus, an early positive component P160 occurs in relation to motion illusion, in agreement with previous results concerning perception of actual motion. When stimuli were presented to produce MAE limited to either the right or left visual hemifield, we also observed a P160 distributed mainly in the right temporal and parietal region. A significant increase in this component was observed in the right posterior temporal region with left hemifield stimulation, while no significant increase was observed with right hemifield stimulation. The right hemispheric dominance of P160 seemed to result partly from functional specialization of the right hemisphere, but hemispheric differences in attentional mechanisms also might contribute to the asymmetric distribution of P160. PMID- 11867253 TI - The experimental combination of rTMS and fMRI reveals the functional relevance of parietal cortex for visuospatial functions. AB - We combined repetitive transcranial magnetic stimulation (rTMS) and functional magnetic resonance imaging (fMRI) to investigate the functional relevance of parietal cortex activation during the performance of visuospatial tasks. fMRI provides information about local transient changes in neuronal activation during behavioural or cognitive tasks. Information on the functional relevance of this activation was obtained by using rTMS to induce temporary regional deactivations. We thereby turned the physiological parameter of brain activity into an independent variable controlled and manipulated by the experimenter and investigated its effect on the performance of the cognitive tasks within a controlled experimental design. We investigated cognitive tasks that were performed on the same visual material but differed in the demand on visuospatial functions. For the visuospatial tasks we found a selective enhancement of fMRI signal in the superior parietal lobule (SPL) and a selective impairment of performance after rTMS to this region in comparison to a control group. We could thus show that the parietal cortex is functionally important for the execution of spatial judgements on visually presented material and that TMS as an experimental tool has the potential to interfere with higher cognitive functions such as visuospatial information processing. PMID- 11867254 TI - Different brain activation patterns during production of animals versus artefacts: a PET activation study on category-specific processing. AB - To study neural correlates of category-specific processing, we measured relative cerebral blood flow changes by PET (oxygen-15) in young healthy subjects while they produced exemplars of animals or artefacts to written subcategory prompts. In comparison to a baseline (word reading), production of animal names elicited increased rCBF in the right inferior temporal region. This fits to recent lesion data on semantic impairment with animals, as well as imaging data on object recognition and semantic retrieval. In our study, it may represent an involvement of visual imagery in generation of animal names. In contrast, production of artefact names elicited increased rCBF in frontoparietal regions previously related to attention and mental effort. PMID- 11867255 TI - Event-related potential features indexing central auditory discrimination by newborns. AB - Behavioral research has produced little evidence on sound feature discrimination in neonates. Sensory processes underlying sound perception can be studied using the mismatch negativity (MMN) component of auditory event-related potentials (ERPs), which is not contingent on conscious perception and response. Thus, MMN is suitable for studying newborns, who are difficult to obtain behavioral responses from. The present study thus utilized spectrally rich sounds, known to elicit the most replicable MMN in adults, to investigate newborns' preattentive analysis of sound duration and frequency changes. An attempt was also made to control for the obligatory ERP effects on the MMN. Three-partial harmonic tones were presented in Duration and in Frequency oddball conditions to 55 newborns. In the other two, Equiprobable duration and Equiprobable frequency, conditions frequency and duration deviants of the oddball paradigms were presented with equal probabilities among sounds of other durations and frequencies. MMN was elicited in 81% of newborns in Frequency oddball condition and in 78% of newborns in Duration oddball condition. No significant amplitude differences between the duration and frequency MMNs were found, but MMN latency was delayed in Duration condition. The obligatory components seemed to contribute significantly to the deviant-standard difference in Duration but not in Frequency condition. The majority of neonates appear to possess effective sound frequency and duration discrimination mechanisms. Their preattentive sound discrimination is facilitated by spectrally rich sound content. The present findings support a change-detection nature of MMN in neonates; however, sound duration-related obligatory effects need to be taken into account in infant MMN studies. PMID- 11867256 TI - The effect of item sequence on brain activity during recognition memory. AB - Two basic types of experimental designs are available to measure the brain indices of successful recognition memory. Both designs have in common that brain activity elicited by correctly recognized studied (old) items is compared to brain activity elicited by correctly rejected new items. In blocked designs, the two item types are presented in separate lists. In random-mixed designs, on the other hand, they are randomly intermixed within the same list. Early studies reported that in random-mixed designs, brain activity can be affected by the sequence in which different types of items are presented. Here, we investigated to what extent such sequence effects also occur in random-mixed designs of recognition memory. Event-related potentials (ERPs) were obtained while subjects made recognition judgments on pseudorandom sequences of visually presented new and old words. Changes in the sequence from one item-type to another (change from old to new and from new to old) were associated with a prominent P300 potential for new words following old words but not vice versa. Subjects with larger P300 potentials made less false alarms to such new words. Successive presentations of the same item-type (sequence 'blocks': three old words in a row and three new words in row) were associated with a left parietal positive slow shift for successive old words but not for successive new words. The data suggest that new words require extensive context-updating if they are presented within the context generated by preceding old words. Due to these sequence effects, the present data question the assumption that random-mixed designs are generally favorable over block designs. PMID- 11867257 TI - Consistency of inter-trial activation using single-trial fMRI: assessment of regional differences. AB - Recently, the technique of single-trial fMRI was introduced, which allows the assessment of hemodynamic responses to single task executions (e.g. sensory, motor, or cognitive). In this study, single-trial fMRI was used to examine regional differences in the inter-trial consistency (ITC) of brain activity related to the processing of a dynamic visuo-spatial imagery task. For every single trial, a t-statistic assessing task-related activation was calculated and thresholded at a p-value of P < or =l0.05 (uncorrected). The percentage of trials with t-values above the threshold was used to assess differences in the consistency of brain activity in occipital, parietal, premotor and prefrontal regions of interest. While most of these regions showed activation which was highly consistent across trials, the consistency of activation was significantly reduced in the dorsolateral prefrontal cortex. We hypothesize that the consistency of activation across trials may be interpreted as an indicator of the functional relevance of a brain region for processing and solving a cognitive task. Thus, the analysis concept presented here has the potential to provide new insights into the neuro-cognitive mechanisms of human information processing. In addition, the results of this study confirm that averaging across trials might result in a significant loss of information about functional neuroanatomy. Regions which are active in some trials only, which show only weak activity increases, or whose activity is not constant and time-locked, might not show up in averaged activation maps and could thus erroneously be interpreted as irrelevant for task processing. PMID- 11867258 TI - Early involvement of the temporal area in attentional selection of grating orientation: an ERP study. AB - The aim of the present study was to investigate the neural mechanisms of stimulus orientation selection in humans by recording event-related potentials (ERPs) of the brain with a 32-channel montage. Stimuli were isoluminant black-and-white gratings (3 cpd) having an orientation of 50, 70, 90, 110 and 130, randomly presented in the foveal portion (2 of visual angle) of the central visual field. The task consisted in selectively attending and responding to one of the five grating orientations, while ignoring the others. ERP results showed that orientation selection affected neural processing starting already at an early post-stimulus latency. The P1 component (80-140 ms) measured at temporal area, which might well be reflecting the activity of the ventral stream (i.e. 'WHAT' system) of the visual pathways, showed an enhanced amplitude for target orientations. These effects increased with progressive neural processing over time as reflected by selection negativity (SN) and P300 components. In addition, both reaction times (RTs) and ERPs showed a strong 'oblique' effect, very probably reflecting the perceptual predominance of orthogonal versus oblique stimulus orientation in the human visual system: RTs were much faster, and SN and P300 components much larger, to gratings presented vertically than in other orientations. PMID- 11867259 TI - DNA methylation of nuclear receptor genes--possible role in malignancy. AB - The members of the nuclear receptor superfamily are known to mediate a wide array of basic biological processes, such as regulation of cell growth and differentiation, and induction of apoptosis. In several human malignancies, this central control function of nuclear receptors is disturbed, which seems to play an important role in tumor development and progression. Many nuclear receptor genes have been reported to be downregulated in malignancies; however, only a few mutations, gene arrangements, deletions or similar genetic changes have been shown to occur in these tumors. During the last decade, increasing attention has been directed towards epigenetic mechanisms of gene regulation such as DNA methylation. Many nuclear receptor genes can be silenced through aberrant methylation in tumors; epigenetic silencing, therefore, represents an additional mechanism that modifies expression of key genes during carcinogenesis. This review will give insights into the role of DNA methylation in the silencing of nuclear receptor genes and its involvement in human malignancies. PMID- 11867261 TI - European sea bass (Dicentrarchus labrax L.) cytochrome P450arom: cDNA cloning, expression and genomic organization. AB - Cytochrome P450arom, a key enzyme in the hormonal steroidogenic pathway, mediates the conversion of androgens to estrogens. This work describes the molecular cloning of the cDNA encoding the European sea bass (Dicentrarchus labrax L.) cytochrome P450arom by means of reverse transcriptase and polymerase chain reaction (RT-PCR) and 5' and 3'-rapid amplification of cDNA ends (RACE) analyses. The cDNA is 1822bp in length and encodes a putative protein of 517 amino acids. Northern blot analysis revealed that the ovary expressed a transcript of about 2.2kb in size. Analysis of the deduced amino acid sequence indicated 62-86% identity with ovarian P450arom of other teleost fish, the highest identity being found with the Japanese flounder, Paralichthys olivaceous. Identity was lower (56 65%) with the P450arom forms first reported in teleost brain. Only 52% identity was observed with the corresponding fragment of the cartilaginous fish, Dasyatis sabina. RT-PCR revealed that the sea bass P450arom mRNA was also expressed, at low levels, in testis and brain. Between the 5' and 3'-untranslated terminal regions (UTR), the sea bass CYP19 gene contains eight introns. All introns conform to the GT/AG rule for RNA splicing and are inserted in exactly the same positions as those found in Oryzias latipes and the human CYP19 gene. PMID- 11867260 TI - Digital expression profiles of the prostate androgen-response program. AB - The androgen receptor (AR) and cognate ligands regulate vital aspects of prostate cellular growth and function including proliferation, differentiation, apoptosis, lipid metabolism, and secretory action. In addition, the AR pathway also influences pathological processes of the prostate such as benign prostatic hypertrophy and prostate carcinogenesis. The pivotal role of androgens and the AR in prostate biology prompted this study with the objective of identifying molecular mediators of androgen action. Our approach was designed to compare transcriptomes of the LNCaP prostate cancer cell line under conditions of androgen depletion and androgen stimulation by generating and comparing collections of expressed sequence tags (ESTs). A total of 4400 ESTs were produced from LNCaP cDNA libraries and these ESTs assembled into 2486 distinct transcripts. Rigorous statistical analysis of the expression profiles indicated that 17 genes exhibited a high probability (P>0.9) of androgen-regulated expression. Northern analysis confirmed that the expression of KLK3/PSA, FKBP5, KRT18, DKFZP564K247, DDX15, and HSP90 is regulated by androgen exposure. Of these, only KLK3/PSA is known to be androgen-regulated while the other genes represent new members of the androgen-response program in prostate epithelium. LNCaP gene expression profiles defined by two independent experiments using the serial analysis of gene expression (SAGE) method were compared with the EST profiles. Distinctly different expression patterns were produced from each dataset. These results are indicative of the sensitivity of the methods to experimental conditions and demonstrate the power and the statistical limitations of digital expression analyses. PMID- 11867262 TI - Vanadate increases glucocorticoid receptor-mediated gene expression: a novel mechanism for potentiation of a steroid receptor. AB - Transition metal oxyanions, such as molybdate, tungstate and vandadate, have been shown to prevent in vitro hormone-induced activation of the glucocorticoid receptor (GR) by blocking dissociation of the GR/heat shock protein heterocomplex. In this work, we report a novel effect of vanadate: in vivo potentiation of GR-mediated gene expression. In cells stably-transfected with complex (mouse mammary tumor virus (MMTV)) or minimal GR-regulated CAT reporters, treatment with 500muM vanadate caused CAT gene expression to dramatically increase, even at saturating concentrations of dexamethasone; while no such effect was seen in response to RU486 antagonist. Similar treatment with molybdate had no effect on GR activity, suggesting that the response to vanadate was not a general property of transition metal oxyanions. Treatment with vanadate after hormone-induced nuclear translocation of the GR also caused potentiation, demonstrating that vanadate was acting on a post-transformation event, perhaps by affecting the transactivation function of DNA-bound GR. Paradoxically, vanadate caused an apparent but temporary "loss" of GR protein immediately after treatment (as measured by loss of reactivity to BuGR2 antibody and of hormone-binding capacity) that returned to normal at approximately 8h post-treatment, suggesting that potentiation of GR transactivation function (as measured by our CAT assays) was probably occurring during the later stages (8-24h) of this assay. However, gel shift analyses revealed that vanadate could induce binding of the hormone free GR to glucocorticoid response element (GRE)-containing oligonucleotides immediately after treatment. Thus, the rapid vanadate-induced "loss" of GR was not due to degradation of GR protein. Yet, vanadate in the absence of hormone had no effect on CAT reporter expression, demonstrating that this form of the GR still requires agonist for its enhanced transcriptional activity. As an indication of the potential mechanism of vanadate action, vanadate was found to dramatically stimulate the mitogen-activated protein kinases, ERK-1 and ERK-2. In addition, vanadate potentiation of GR reporter gene expression was completely blocked by the tyrosine kinase inhibitor herbimycin A. Taken as a whole, our results suggest that vanadate can have dramatic and complex effects on GR structure and function, resulting in hormone-free activation of GR DNA-binding function, as well as alterations to the BuGR2 epitope and hormone-binding domains -while at the same time stimulating tyrosine phosphorylation pathways controlling GR-mediated gene transcription. PMID- 11867263 TI - Oestrogenic activity of parabens in MCF7 human breast cancer cells. AB - Parabens (4-hydroxybenzoic acid esters) have been recently reported to have oestrogenic activity in yeast cells and animal models. Since the human population is exposed to parabens through their widespread use as preservatives in foods, pharmaceuticals and cosmetics, we have investigated here whether oestrogenic activity of these compounds can also be detected in oestrogen-sensitive human cells. We report on the oestrogenic effects of four parabens (methylparaben, ethylparaben, n-propylparaben, n-butylparaben) in oestrogen-dependent MCF7 human breast cancer cells. Competitive inhibition of [3H]oestradiol binding to MCF7 cell oestrogen receptors could be detected at 1,000,000-fold molar excess of n butylparaben (86%), n-propylparaben (77%), ethyl-paraben (54%) and methylparaben (21%). At concentrations of 10(-6)M and above, parabens were are able to increase expression of both transfected (ERE-CAT reporter gene) and endogenous (pS2) oestrogen-regulated genes in these cells. They could also increase proliferation of the cells in monolayer culture, which could be inhibited by the antiestrogen ICI 182,780, indicating that the effects were mediated through the oestrogen receptor. However, no antagonist activity of parabens could be detected on regulation of cell proliferation by 17 beta-oestradiol at 10(-10)M. Molecular modelling has indicated the mode by which paraben molecules can bind into the ligand binding pocket of the crystal structure of the ligand binding domain (LBD) of the oestrogen receptor alpha (ERalpha) in place of 17beta-oestradiol; it has furthermore shown that two paraben molecules can bind simultaneously in a mode in which their phenolic hydroxyl groups bind similarly to those of the meso hexoestrol molecule. Future work will need to address the extent to which parabens can accumulate in hormonally sensitive tissues and also the extent to which their weak oestrogenic activity can add to the more general environmental oestrogen problem. PMID- 11867264 TI - Effects of the environmental estrogens bisphenol A, o,p'-DDT, p-tert-octylphenol and coumestrol on apoptosis induction, cell proliferation and the expression of estrogen sensitive molecular parameters in the human breast cancer cell line MCF 7. AB - In the presented study, we have analysed effects of the environmental estrogens bisphenol A (BPA), p-tert-octylphenol (OCT), o,p'-DDT (DDT) and coumestrol (COU) on cell proliferation, apoptosis induction, progesterone receptor (PR) and androgen receptor (AR) mRNA expression and ER alpha protein expression in comparison to estradiol (E2) and the selective ER modulator (SERM) raloxifene (RAL) and the pure antiestrogen faslodex (ICI 182780) in the human breast cancer cell line MCF-7. A dose dependent analysis of the cell cycle distribution of MCF 7 cells after administration of OCT, DDT and COU revealed a significant induction of cell proliferation and reduced rate of apoptosis. Maximum induction of cell proliferation and the lowest rate of apoptosis could be observed at a dose of 10( 6)M. Interestingly, administration of BPA reduces the rate of apoptosis, but does not enhance proliferation at any dose analysed. PR mRNA expression in MCF-7 cells was up regulated after administration of COU and DDT, whereas treatment with BPA and OCT did not effect PR mRNA expression. AR mRNA expression was down regulated by COU, but not effected by BPA, DDT and OCT. The expression of ER alpha protein in the breast cancer cells was slightly down regulated by COU and DDT, but unaffected by BPA and OCT. In summary and in comparison to the effects observed after administration of E2, RAL and ICI our data indicate that none of the analysed compounds exhibit properties comparable to RAL and ICI. COU and DDT exhibit properties which are very similar to E2. Administration of BPA and OCT did not effect any of the estrogen sensitive molecular parameters analysed. Nevertheless OCT is a very potent stimulator of cell proliferation in MCF-7 cells. Surprisingly, BPA is not able to induce the proliferation of MCF-7 breast cancer cells, but turns out to be a very potent inhibitor of apoptosis. For this reason and in agreement to the effects of BPA on the molecular parameters analysed, we conclude that BPA does not act in a classical estrogen like manner in MCF-7 breast cancer cells. PMID- 11867265 TI - Assessment of the ability of type 2 cytochrome b5 to modulate 17,20-lyase activity of human P450c17. AB - The 17 alpha-hydroxylase and 17,20-lyase activities of P450c17 lead to the production of 17 alpha-hydroxypregnenolone (17 alpha-OH-Preg) and dehydroepiandrosterone (DHEA), respectively, in different tissues. The mechanisms of differential regulation of these two activities are not yet fully elucidated. It has been previously shown that cytochrome b5 (cyt-b5) could facilitate the 17,20-lyase activity of human P450c17. Recently, a cDNA (type 2 cyt-b5) sharing 45.8% homology with type 1 cyt-b5 has been isolated from human testis. Since high 17,20-lyase activity is required for the production of androgens in the testis, we wanted to determine the importance of this second cDNA in the modulation of P450c17 17,20-lyase activity and hence, its role in the formation of active androgens. We therefore isolated type 2 cyt-b5 from human testis by RT-PCR and analyzed, by transient transfection in transformed human embryonic kidney cells (HEK-293) of various amounts of vectors expressing cyt-b5, P450-reductase and P450c17, its ability to modulate the 17,20-lyase activity of human P450c17. Results show that, in the presence of NADPH cytochrome P450 reductase (P450-red), type 2 cyt-b5 increases 17,20-lyase activity to a level comparable to that of type 1. These results support the idea that types 1 and 2 cyt-b5 could be involved in the differential modulation of 17 alpha-hydroxylase and 17,20-lyase activities of P450c17. Furthermore, the analysis of mRNA expression of types 1 and 2 cyt-b5 by RT-PCR using primers specific to each type showed that both types are present in the liver but also in the adrenal and testis. PMID- 11867266 TI - Expression of Bcl-2, Bcl-x, Mcl-1, Bax and Bak in human uterine leiomyomas and myometrium during the menstrual cycle and after menopause. AB - To investigate the expression of Bcl-2, Bcl-x, Mcl-1, Bax and Bak proteins in human uterine leiomyomas and homologous myometrium during the menstrual cycle and after menopause. The expression of Bcl-2, Bcl-x, Mcl-1, Bax and Bak in leiomyomas (n=24) and myometrial samples (n=22) from women with leiomyomas was measured by immunohistochemistry and Western blot. Measured by immunohistochemistry, a significant difference between leiomyomas and myometrium was observed only for the Bax protein, in tissues obtained from women in the secretory phase of the menstrual cycle. The Bcl-2 staining was more abundant in leiomyomas than in myometrium only in tissues obtained in the proliferative phase of the cycle. Bcl 2 was more abundant in leiomyomas from women of fertile age than in leiomyomas from menopausal women. No significant differences were observed for the Bcl-x or Bak proteins, whereas the Mcl-1 protein was significantly less abundant in secretory phase leiomyomas than in leiomyomas from menopausal women. Western blot analysis based on pools of tissue extracts from the different groups essentially confirmed the data obtained by immunohistochemistry. Bcl-2 family proteins are expressed in leiomyomas and myometrium in different phases related to and influenced by gonadal steroids. These proteins are suggested to interact with each other in the regulation of programmed cell death, apoptosis, but their specific role in growth control of uterine leiomyomas remains to be investigated. PMID- 11867267 TI - Inhibition of MEK1,2/ERK mitogenic pathway by estrogen with antiproliferative properties in rat aortic smooth muscle cells. AB - Proliferation and migration of vascular smooth muscle cells (VSMCs) are believed to contribute significantly to intimal thickening in atheroscleosis, restenosis, and venous bypass graft disease. Estrogen inhibits proliferation and migration of VSMCs. However, antiproliferative mechanisms of estrogen were not well elucidated yet. In this study, we investigated the antiproliferative effect of estrogen to determine whether the transduction signals and protooncogenes were affected in rat aortic smooth muscle cells (RASMCs). Estrogen inhibited the proliferative response stimulated by 5% fetal bovine serum (FBS) dose-dependently in RASMCs (IC50: 40 nM). In 0.5% serum-treated RASMCs, estrogen dramatically inhibited the activity of extracellular signal-regulated kinases (ERK) followed by inhibition of MEK1,2 activity in dose-dependent manner without affecting the other mitogen activating protein kinases (MAPKs), c-jun N-terminal kinases (JNK) and p38. Induction of Elk-1 mRNA was significantly reduced dose-dependently up to 100 nM of estrogen. These results indicate that the antiproliferative effects of estrogen in RASMCs involved ERK inhibition followed by the inactivation of MEK1,2 and downregulation of Elk-1 expression. PMID- 11867268 TI - Hypothalamic 5 alpha-reductase and 3 alpha-oxidoreductase activity in the male rat. AB - The aim of the present study was to assess the progesterone (Pr) transforming 5 alpha-reductase (5 alpha-R) and 3 alpha-oxidoreductase (3 alpha-OR) activities in the hypothalamus of the male rat as a function of age and following castration and/or adrenalectomy performed at the sixth day of life. The hypothalamic activity of these enzymes was estimated from the sum of the 5 alpha- or 3 alpha reduced metabolites produced from 14C-labeled Pr incubated "in vitro" with hypothalamic tissue. Plasma levels of testosterone (T), progesterone (Pr), estrone (E1), luteinizing hormone (LH) and follicle stimulating hormone (FSH) were measured simultaneously. Special attention was paid to the GC/MS analysis of the endogenous content of the hypothalamic Pr-metabolites 3 alpha-hydroxy-pregn-4 en-20-one (3 alpha-Pr), 5 alpha-pregnane-3,20-dione (5 alpha-Pr) and 3 alpha hydroxy-5 alpha-pregnan-20-one (5 alpha,3 alpha-Pr). The high 5 alpha-R and 3 alpha-OR activities estimated in the hypothalamus of prepubertal rats are not related to the action of gonadal or adrenal steroids. Substantial and comparable endogenous 3 alpha- and/or 5 alpha-Pr-metabolites were found in hypothalami from both prepubertal and mature rats. The results of the present study do not provide evidence for a contributory role of the 3 alpha-hydroxylated Pr derivative to the regulation of gonadotropin secretion in the male rat. PMID- 11867269 TI - Comparison of the hamster and human adrenal P450c17 (17 alpha-hydroxylase/17,20 lyase) using site-directed mutagenesis and molecular modeling. AB - In order to understand the activity specificity of the hamster cytochrome P450 17 alpha-hydroxylase/17,20-lyase (P450c17), we have studied its structure/activity using three hamster P450c17 recombinant mutants (T202N/D240N/D407H). In transiently transfected COS-1 cells, the mutation T202N reduced 17 alpha hydroxylation of pregnenolone and progesterone to 24 and 44% of wild type (WT), respectively, followed by reduced 17,20-cleavage to 71 and 67%, respectively. On the other hand, the mutation D240N decreased specifically 17,20-lyase activity to 61% of WT when incubated with pregnenolone while the mutation D407H only decreased 17 alpha-hydroxylation to 46% when incubated with progesterone.To comprehend the altered activity profiles of these hamster P450c17 mutants, we have elaborated a 3D model of the hamster P450c17 and compared it to our preceding model of the human P450c17. Analysis of the mutants with this model showed that, without direct contact to the substrates, these mutations transmit structural changes to the active site. By analogy, these results support the concept that any cellular changes modifying the external structure of P450c17, such as phosphorylation, could have influence on its active site and enzymatic activities. PMID- 11867270 TI - Evaluation of potential implication of membrane estrogen binding sites on ERE dependent transcriptional activity and intracellular estrogen receptor-alpha regulation in MCF-7 breast cancer cells. AB - The potential involvement of membrane estrogen binding sites in the induction of ERE-dependent transcriptional activity as well as in the regulation of intracellular estrogen receptor alpha (ER-alpha) level under estradiol (E2) stimulation was investigated. Our approach relied upon the use of two DCC-treated E2-BSA (bovine serum albumin) solutions (E2-6-BSA and E2-17-BSA). The absence of detectable free E2 in these solutions was established. Both E2-BSA conjugates led to a transient dose-dependent stimulation of the expression of ERE-luciferase (LUC) reporter gene in MVLN cells (MCF-7 cells stably transfected with a pVit-tk LUC reporter plasmid), a property not recorded with free E2, which maintained enhanced transcriptional activity during the whole experiment. A very low concentration of E2 (10 pM) synergistically acted with E2-BSA conjugates. Hence, ERE-dependent transcriptional activity induced by these conjugates appeared to result from their known interactions with membrane estrogen binding sites. Anti estrogens (AEs: 4-OH-TAM and RU 58,668), which antagonize genomic ER responses, abrogated the luciferase activity induced by E2-BSA conjugates, confirming a potential relationship between membrane-related signals and intracellular ER. Moreover, induction of luciferase was recorded when the cells were exposed to IBMX (3-isobutyl-1-methylxanthine) and cyclic nucleotides (cAMP/cGMP), suggesting the implication of the latter in the signal transduction pathway leading to the expression of the reporter gene. Growth factors (IGF-I, EGF and TGF-alpha) also slightly stimulated luciferase and synergistically acted with 10 pM E2, or 1 microM E2-BSA conjugates, in agreement with the concept of a cross-talk between steroids and peptides acting on the cell membrane. Remarkably, E2-BSA conjugates, IBMX and all investigated growth factors failed to down-regulate intracellular ER in MCF-7 cells, indicating the need for a direct intracellular interaction of the ligand with the receptor to regulate its level. ER elimination was, however, found in the presence of conditioned media (CMs) prepared from cells pre-exposed to E2-BSA conjugates, suggesting that they may produce (a) modulator(s) that may enhance receptor down-regulation when released within the medium. PMID- 11867271 TI - Antiestrogenic activities of Cimicifuga racemosa extracts. AB - Despite the wide use of extracts from the rhizome of black cohosh (Cimicifuga racemosa) for the treatment of menopausal complaints, surprisingly little is known on their potential estrogenic properties, e.g. on estrogen dependent gene transcription. In addition, available informations on the effects on cell proliferation are contradictory. We therefore, tested for estrogenic and antiestrogenic effects of Cimicifuga racemosa extracts on proliferation of MCF-7 cells and on gene expression using ethanolic and iso-propanolic extracts of this medical plant. Estrogenic properties of plant extracts could neither be detected in proliferation assays, nor on gene expression using an estradiol-inducible yeast assay or the estrogen-inducible MVLN cells. In contrast, in all three experimental systems Cimicifuga racemosa antagonized estradiol induced activities. Estradiol induced stimulation of proliferation was inhibited by a dosage >1 microg/ml of extract concentration, gene expression was suppressed by doses of 100-1000 microg/ml of Cimicifuga racemosa extracts. From these results we conclude, that extracts from the rhizome of Cimicifuga racemosa contain compounds with antiestrogenic properties. PMID- 11867272 TI - Conversion of liposomal 4-androsten-3,17-dione by A. simplex immobilized cells in calcium pectate. AB - Arthrobacter simplex ATCC 6946 free and immobilized cells were assayed for their ability to convert 4-androsten-3,17-dione (AD) to 1,4-androstadien-3,17-dione (ADD) in aqueous and liposomal media. Bioconversions were carried out in a 100 ml flask containing 25 ml of AD liposomal or aqueous medium for 3h, and AD concentrations ranging from 0.3 to 1.0 mM were tested. AD/ADD ratios in samples were determined by HPLC. Biotransformation of substrate entrapped in multilamellar vesicles (MLV) was demonstrated to be better than the corresponding free form. In the former case, 2h were necessary to completely bioconvert 1 mM AD. By contrast, 3h were needed to reach 50% bioconversion in (4%) ethanol medium containing 0.63 mM AD. The liposomal medium allows us to perform steroid conversions at high concentrations of AD, reusing immobilized cells in suitable conditions which are non-toxic for microorganisms. PMID- 11867273 TI - From molecular and cellular to integrative heat defense during exposure to chronic heat. AB - Heat acclimation induces adaptive changes that improve the ability to cope with extreme environmental heat. Acclimatory homeostasis is manifested by an expanded dynamic thermoregulatory span (TRS), reflected in the intact organism by a lower temperature threshold (T(sh)) for heat dissipation, and delayed T(sh) for thermal injury. This principle shares common adaptive features with each of the thermoregulatory effectors. In the splanchnic circulation, e.g. the TRS of the thermally induced vasomotor response increases due to greater cardiac output distribution to the splanchnic vasculature, thereby increasing circulatory reserves and delaying thermal injury. During short-term heat acclimation (STHA), accelerated autonomic excitability plays a major role in the control of body temperature. Acclimatory homeostasis, however, is achieved only following long term heat acclimation (LTHA), and is characterized by increased thermal effector efficiency, namely [effector organ output/autonomic signal] ratio >1. Two acclimatory responses, derived from our data on the acclimating rat model, are discussed: (1) acclimation of the cholinergic-muscarinic signaling for water secretion in the submaxillary gland; and (2) acclimatory mechanisms for increased contractile efficiency in the heart. Our data indicate that increased efficiency upon LTHA develops by reprogramming of gene expression. A reduced thyroid hormone level is responsible for some of the molecular adaptive cascades. Delayed thermal injury observed upon acclimation is due to enhanced cytoprotective mechanisms of which the inducible heat shock protein (HSP) 72 kDa plays a major role. Our data indicate that heat acclimation predisposes the HSP molecular machinery to respond faster and increases the constitutive level of the protein. STHA is the time window during which most LTHA adaptations are switched on. PMID- 11867274 TI - Adrenoceptor and local modulator control of cutaneous blood flow in thermal stress. AB - Blood flow to the skin is controlled by body temperatures in two ways: core and mean skin temperature combine in the central nervous system to form a reflex mechanism that controls the frequency of activity in sympathetic nerves to the cutaneous blood vessels; and local mechanisms independent of reflex effects control contractile response to the sympathetic transmitter norepinephrine (NE) at different temperatures. Cutaneous vessels differ in responsiveness to NE across temperatures: in limbs and tails, the superficial vessels constrict more strongly to NE when cooled, while the deep vessels show weaker responses to NE when cooled. This allows the limb to dissipate heat when warm and to conserve heat when cool. The mechanism for this difference in thermal response of deep and superficial vessels is not completely known, but may relate to differences in the adrenoceptors on which NE acts, and/or to the actions of locally produced substances that modulate the responses to NE in different ways at different temperatures. This paper discusses the alpha1- and alpha2-adrenoceptors involved in contraction of deep and superficial cutaneous vessels and also describes the roles of the local modulator nitric oxide, which interacts with adrenoceptors to affect cutaneous blood flow. PMID- 11867276 TI - Lessons from nature: the role of sugars in anhydrobiosis. AB - A review of the role of sugars in anhydrobiosis is presented. PMID- 11867275 TI - Cooling by cutaneous water evaporation in the heat-acclimated rock pigeon (Columba livia). AB - The present study provides an up-to-date overview of the cutaneous water evaporation cooling mechanism in the rock pigeon. Cutaneous water evaporation fully replaces the classic respiratory cooling mechanism in the resting, heat acclimated bird, and is more economical in terms of water conservation. It enables the pigeon to maintain homeostasis, and to breed successfully in harsh environments. Adrenergic signaling is involved in the initiation of this novel mechanism, either by deactivation of the beta-adrenergic receptors (ARs), or activation of the alpha-AR. The adrenergic signaling results in a marked increase in cutaneous blood flow and in the arterial-to-venous blood-flow ratio. This is associated with alterations in the cutaneous capillary wall ultrastructure, which increase its permeability to plasma proteins and water. The end result of this process might be an increase in water efflux from the capillary lumen. The properties of beta-ARs were measured in the cardiac muscle of thermal-acclimated pigeons. Significant down-regulation in the density of beta-ARs, associated with increased affinity of these receptors, was measured in the heat-acclimated pigeon. Concomitantly, changes in the skin ultrastructure and lipid composition were found in very well defined patches in the epidermis of heat-acclimated pigeons. These suppress the skin resistance to water transfer. We suggest that this cooling mechanism involves finely orchestrated adjustments in the ultrastructure of the skin and the cutaneous capillaries, and in skin blood flow. Adrenergic signals are among those factors that regulate this cooling mechanism during exposure to a hot environment. PMID- 11867277 TI - Looking beyond sugars: the role of amphiphilic solutes in preventing adventitious reactions in anhydrobiotes at low water contents. AB - Plants and animals that can survive dehydration accumulate high concentrations of disaccharides in their cells and tissues during desiccation. These sugars are necessary both for the depression of the membrane phase transition temperature of the dry lipid and for the formation of a carbohydrate glass. In the past decade, however, it has become clear that certain types of adventitious enzymatic reactions are possible at low water contents, which along with free-radical mediated damage, can cause hydrolysis of lipids and loss of membrane barrier function. Disaccharides do not necessarily prevent these types of reactions, which suggests that other compounds might also be necessary for protecting organisms from this type of degradation during anhydrobiosis. Arbutin, one possible example, accumulates in large quantities in certain resurrection plants and has been shown to inhibit phospholipase A(2) activity at low water contents. The direct effect of arbutin on membranes under stress conditions depends on the membrane lipid composition. It can serve a protective function during desiccation or freeze/thaw-induced stress in the presence of nonbilayer-forming lipids or a disruptive function in their absence. Other possible amphiphiles, including certain naturally occurring flavonols, may serve as anti-oxidants and some might have similar lipid composition-dependent effects. Such compounds, therefore, are likely to be localized near specific membranes, where they might provide the greatest benefit at the least liability to the organism. PMID- 11867278 TI - The role of amphiphiles. AB - This paper reviews our work on the partitioning of amphiphilic compounds from the cytoplasm into membranes during drying of plant systems, and discusses how relevant this phenomenon might be for anhydrobiosis. Amphiphilic guest molecules do partition into membranes and oil bodies, as demonstrated by the results of in vivo electron paramagnetic resonance spectroscopy on incorporated spin probes. Arguments for the likelihood of endogenous cytoplasmic amphiphiles behaving similarly during dehydration and rehydration of plant systems are presented. Negative and positive aspects of the partitioning are summarized. Positive aspects are the automatic insertion of amphiphilic antioxidants into membranes of the dehydrating organism, and the control of membrane fluidity and the phase transition temperature. A negative aspect is the perturbation of membrane structure, leading to increased permeability and loss of function. The finding that after an initial fluidization during dehydration, the membrane surface becomes immobilized in desiccation-tolerant systems and not in desiccation sensitive systems, is discussed in the light of a strict control of the effect of partitioning. The adaptive significance of amphiphile partitioning into the membranes of anhydrobiotes is discussed. PMID- 11867279 TI - From anhydrobiosis to freeze-drying of eukaryotic cells. AB - Using what has been learned from nature, it has become possible to stabilize biological structures, including intact cells, in the dry state. Stabilization of cells or tissues in the dried state is of considerable practical significance, as is described in this review. The need for stabilization of cells in the dried state is particularly urgent in bloodbanks, where proper storage of blood cells (platelets and erythrocytes) is still a major problem. Human blood platelets are stored in blood banks for 5 days, after which they are discarded according to Federal regulation. This short lifetime has led to a chronic shortage of platelets. We report here that platelets can be preserved by freeze-drying them with trehalose, a sugar found at high concentrations in organisms that naturally survive drying. We suggest that this finding will obviate the storage problem with platelets and that the principles established here may be extended to more complex eukaryotic cells. PMID- 11867280 TI - Membrane behavior as influenced by partitioning of amphiphiles during drying: a comparative study in anhydrobiotic plant systems. AB - During cellular desiccation, reduction in volume can in principle cause amphiphilic compounds to partition from the cytoplasm into membranes, with structural perturbance as the result. Here, we studied the effect of partitioning of endogenous amphiphiles on membrane surface dynamics in desiccation-tolerant and -intolerant, higher and lower plant systems, using electron paramagnetic resonance (EPR) spin probe techniques. Labeling cells with the amphiphilic spin probe perdeuterated TEMPONE (PDT) enabled partitioning into the various phases to be followed. During drying, PDT molecules preferentially partitioned from the aqueous cytoplasm into the membrane surface and, at advanced stages of water loss, also into oil bodies. There was no specific partition behavior that could be correlated with lower/higher plants or with desiccation-tolerance. In vivo labeling with 5-doxylstearate (5-DS) enabled membrane surface fluidity to be characterized. In hydrated plants, the 5-DS spectra contained an immobile and a fluid component. The characteristics of the immobile component could not be specifically correlated with either lower or higher plants, or with desiccation tolerance. The relative contribution of the fluid component to the 5-DS spectra was higher in lower plants than in higher plants, but considerably decreased with drying in all desiccation-tolerant organisms. In contrast, the proportion of the fluid component in desiccation-sensitive wheat seedling root was higher than that in desiccation-tolerant wheat axis and considerably increased at the onset of water loss. We suggest that partitioning of amphipaths fluidize the membrane surface, but that in desiccation-tolerant systems the membranes are protected from excessive fluidization. PMID- 11867281 TI - The Antarctic Psychrobacter sp. TAD1 has two cold-active glutamate dehydrogenases with different cofactor specificities. Characterisation of the NAD+-dependent enzyme. AB - Psychrobacter sp. TAD1 is a psychrotolerant bacterium from Antarctic frozen continental water that grows from 2 to 25 degrees C with optimal growth rate at 20 degrees C. The new isolate contains two glutamate dehydrogenases (GDH), differing in their cofactor specificities, subunit sizes and arrangements, and thermal properties. NADP+-dependent GDH is a hexamer of 47 kDa subunits and it is comparable to other hexameric GDHs of family-I from bacteria and lower eukaria. The NAD+-dependent enzyme, described in this communication, has a subunit weight of 160 kDa and belongs to the novel class of GDHs with large size subunits. The enzyme is a dimer; this oligomeric arrangement has not been reported previously for GDH. Both enzymes have an apparent optimum temperature for activity of approximately 20 degrees C, but their cold activities and thermal labilities are different. The NAD+-dependent enzyme is more cold active: at 10 C it retains 50% of its maximal activity, compared with 10% for the NADP+-dependent enzyme. The NADP+-dependent enzyme is more heat stable, losing only 10% activity after heating for 30 min, compared with 95% for the NAD+-dependent enzyme. It is concluded that in Psychrobacter sp. TAD1 not only does NAD+-dependent GDH have a novel subunit molecular weight and arrangement, but that its polypeptide chains are folded differently from those of NADP+-dependent GDH, providing different cold-active properties to the two enzymes. PMID- 11867282 TI - Generation and application of chicken egg-yolk antibodies. AB - Despite the fact that the use of chicken as immunization host brings many advantages to the production of polyclonal antibodies, the generation of egg yolk immunoglobulins (IgY) is rarely chosen. In this review, we report on the fast and efficient method for generation and affinity purification of IgY, in this case raised against the alpha-subunit of hypoxia-inducible factor-1 (HIF-1). The IgY antibody was successfully applied in a variety of methods and a number of different species for HIF-1alpha detection. In electrophoretic mobility shift assays, the IgY antibody recognized the native HIF-1 complex. The IgY antibody also detected HIF-1alpha protein on Western blots with extracts derived from human, monkey, pig, dog and mouse cell lines grown under hypoxic conditions. Immunofluorescence and immunoprecipitation experiments using the IgY antibody allowed detection and subcellular localization of HIF-1alpha in the nuclei of hypoxic cells. Chicken antibody production brings great benefit concerning the welfare of the immunized animals, due to non-invasive antibody harvesting with the added convenience of simple egg collection. An additional advantage is the fast and simple IgY isolation from egg yolk. IgY technology is a great improvement and should be considered as a good alternative to conventional polyclonal antibody production in mammals. PMID- 11867283 TI - Fish at high pressure: a hundred year history. AB - Two main periods can be considered in the history of fish metabolism under pressure. The first period (roughly from 1870 to 1970) was mainly descriptive: survival times and behavior were studied and some authors described an increase in oxygen consumption under pressure; later, the counteracting effects of high temperature on pressure were mentioned. The second period (from 1970 onwards) was more integrative and two major ways were explored. The first was to use shallow water fish, experimentally exposed to hydrostatic pressure, which can induce a metabolic state resembling histotoxic hypoxia. The second way was to use deep living fish which have, when compared to surface fish, muscle enzymes with higher structural stability, lower activity (in relationship with habitat depth) and kinetics that are less sensitive to pressure increase. Using this approach, it was also shown that muscle composition and function were somewhat different at depth and that deep fish are well adapted to pressure partly by maintaining membrane fluidity (homeoviscous theory). Since about 1990, the two above mentioned approaches have still been pursued but by fewer researchers. Studies on deep-living fish are mainly concerned with enzyme kinetics whereas shallow water fish are used mainly for cellular energetic studies. Regarding this topic, it has been shown that yellow freshwater eels are able to acclimate to high-pressure effects, by optimizing membrane fluidity and composition (as achieved by deep living fish), by improving oxidative phosphorylation (increase of P/O ratio) and the glycolytic pathway. PMID- 11867284 TI - Ion channels under high pressure. AB - Hydrostatic pressure (<100 MPa) affects the kinetics of ion channels but not their conductance. In voltage-gated channels, pressure acts on the movement of the charge sensor and on the conformational change involved in opening the channel pore. It has also been shown to act on N-type inactivation ball-binding, C-type inactivation and to activate BK channels. There is little doubt that these are sites of adaptation to high pressure in the channels of deep-sea animals. Pressure studies should not be regarded in isolation; they relate well to experiments using other variables such as osmotic pressure, solvent viscosity and temperature. Furthermore ion channels could transduce pressure in the sensory system of aquatic animals, providing information about the animals' depth, a prediction supported by our knowledge of heat-activated channels in mammals. PMID- 11867285 TI - Influence of activation and germination on high pressure inactivation of ascospores of the mould Eurotium repens. AB - We investigated heat activation and germination of Eurotium repens ascospores to follow high pressure inactivation. Activation energy and entropy values strengthen the idea of protein denaturation as the underlying mechanism of heat activation. Preceding activation, germination or a combination of both affected high pressure inactivation in different ways. Activation followed immediately by high pressure treatment led to the most efficient improvement in inactivation. However, a pause after activation caused a partial re-establishment of the spores' stability and less efficient high pressure inactivation. Germination stabilized the spores against high pressure. A combined treatment of activation and germination led to an initially fast inactivation, but compared to high pressure treatment of only activated spores the time course of inactivation was slowed down. PMID- 11867286 TI - Air exposure and functionality of Chamelea gallina haemocytes: effects on haematocrit, adhesion, phagocytosis and enzyme contents. AB - The Venus clam Chamelea gallina is fairly common along the western coasts of the Adriatic and is subjected to intense fishing. Since over the last 20 years extensive hypoxic and anoxic conditions have repeatedly damaged this natural resource, we decided to study the effects of anoxic stress on the functionality of clam haemocytes and the consequences on immune responses. Clams, exposed to air, close their valves and tissues become anoxic and metabolism processes switch to anaerobiosis. In these conditions, a significant decrease in the haematocrit value and in the percentage of acid phosphatase-positive haemocytes was observed, while the number of cells with beta-glucuronidase significantly increased after day 1. The above indices generally returned to control values when clams were re immersed in seawater after 1 day of treatment. Clams exposed to air for 2 days and then re-immersed, attempted to recover in the subsequent 3 days. Animals had fully recovered on day 4. Three-day-exposed clams did not recover. Phagocytic and adhesion indices decreased significantly after the first day of air exposure. The change in frequency of three types of circulating cells (spreading, round, apoptotic) was also monitored. PMID- 11867288 TI - Free amino acids in claw muscle and haemolymph from Australian freshwater crayfish at different stages of the moult cycle. AB - Amino acids were measured in claw muscle and haemolymph in the freshwater decapod crustacean, Cherax destructor, at different stages of the moult cycle. The total pool of amino acids in muscles from animals in intermoult (97+/-13 mmol kg(-1) muscle), premoult (80+/-20 mmol kg(-1)) and postmoult (97+/-19 mmol kg(-1)) were not significantly different. Despite the relatively stable total pool of amino acids, there were changes in the concentrations of alanine, glutamine and proline over the moult cycle. Compared to intermoult, claw muscles from animals in premoult had a lower concentration of proline, and animals in postmoult had higher concentrations of alanine and glutamine, but lower concentrations of proline. Concentrations of alanine and glutamine in claw muscle of animals in postmoult were higher and proline concentrations lower than in the same animals during the premoult stage. The concentration of proline in haemolymph was lower in animals in premoult and postmoult compared to intermoult. The total amino acid pool in the claw muscle of Cherax destructor did not change significantly over the moult which is distinctly different to the changes in amino acids reported in the claw muscles of marine decapod crustaceans. PMID- 11867287 TI - Anoxic survival potential of bivalves: (arte)facts. AB - The anoxic survival time of the bivalves Chamelea gallina, Cerastoderma edule and Scapharca inaequivalvis from two different ecosystems and differing anoxia tolerances was studied in static (closed) and flow-through systems. The antibiotics chloramphenicol, penicillin and polymyxin were added, and molybdate (specific inhibitor of the process of sulfate reduction). Survival in (near) anoxic seawater of Chamelea was studied in a static system by comparing untreated seawater with autoclaved seawater and untreated clams with clams incubated in well-aerated seawater, containing the broad-spectrum antibiotic chloramphenicol, prior to the anoxic survival test. With untreated clams and natural seawater (median mortality time 2.4 days) a decrease in pH and exponential accumulation of sulfide and ammonium was observed in the anoxic medium, indicating excessive growth of (sulfate reducing) bacteria. In sterilized seawater LT50 (2.1 days) was not significantly different and again considerable amounts of ammonium and sulfide accumulated. However, pre-treatment of clams with chloramphenicol resulted in an increase of LT50 (11.0 days) by approximately fivefold. Accumulation of ammonium and sulfide was retarded, but was finally even stronger than in the medium containing untreated clams. Median mortality times were 2.5 and 2.4 days for Chamelea and 2.7 and 2.9 days for Cerastoderma for static and flow-through incubations, respectively. Addition of chloramphenicol increased strongly survival time in both systems with corresponding values of 11.0 and 16.3 days for Chamelea, and 6.4 and 6.5 days for Cerastoderma. LT50 of Scapharca in anoxic seawater was 14.4 days. Chloramphenicol and penicillin increased median survival time to 28.5 and 28.7 days, respectively, whereas polymyxin displayed no effect (LT50=13.6 days). Molybdate added to artificial sulfate free seawater blocked biotic sulfide formation, but did not improve survival time (LT50=13.7 days). Overall the results indicate that proliferation of anaerobic pathogenic bacteria, firmly associated with the bivalves, is a main cause of death besides lack of oxygen. Bacterial damage is probably caused by injury of the tissues of the clams and not by the release of noxious compounds to the medium. PMID- 11867289 TI - Effects of anaesthesia on blood gases, acid-base status and ions in the toad Bufo marinus. AB - It is common practice to chronically implant catheters for subsequent blood sampling from conscious and undisturbed animals. This method reduces stress associated with blood sampling, but anaesthesia per se can also be a source of stress in animals. Therefore, it is imperative to evaluate the time required for physiological parameters (e.g. blood gases, acid-base status, plasma ions, heart rate and blood pressure) to stabilise following surgery. Here, we report physiological parameters during and after anaesthesia in the toad Bufo marinus. For anaesthesia, toads were immersed in benzocaine (1 g l(-1)) for 15 min or until the corneal reflex disappeared, and the femoral artery was cannulated. A 1 ml blood sample was taken immediately after surgery and subsequently after 2, 5, 24 and 48 h. Breathing ceased during anaesthesia, which resulted in arterial Po(2) values below 30 mmHg, and respiratory acidosis developed, with arterial Pco(2) levels reaching 19.5+/-2 mmHg and pH 7.64+/-0.04. The animals resumed pulmonary ventilation shortly after the operation, and oxygen levels increased to a constant level within 2 h. Acid--base status, however, did not stabilise until 24 h after anaesthesia. Haematocrit doubled immediately after cannulation (26+/ 1%), but reached a constant level of 13% within 24 h. Blood pressure and heart rate were elevated for the first 5 h, but decreased after 24 h to a constant level of approximately 30 cm H2O and 35 beats min(-1), respectively. There were no changes following anaesthesia in mean cellular haemoglobin concentration, [K+], [Cl-], [Na+], [lactate] or osmolarity. Toads fully recovered from anaesthesia after 24 h. PMID- 11867290 TI - Interrelationships among plasma progesterone concentrations, luteal anatomy and function, and placental ontogeny during gestation in a viviparous lizard (Niveoscincus metallicus: Scincidae). AB - Plasma progesterone concentrations were measured at six stages of gestation in the viviparous lizard Niveoscincus metallicus. Anatomical and functional parameters of luteal activity were also investigated. The diameter of the corpus luteum (CL) decreased gradually though gestation, as did the diameter of the luteal cells. Major degenerative changes were observed in CLs post-partum. Plasma progesterone concentrations were basal both prior to, and just after, ovulation; a rapid increase occurred in early gestation. Plasma progesterone concentrations remained elevated until late gestation, but fell some 2 weeks before parturition. In vitro production of progesterone was greater in CLs in mid- than in late gestation, and the addition of prostaglandin F(2alpha) to the incubation medium had no effect on progesterone production. Non-luteal ovarian tissue and adrenals produced progesterone, but at approximately one-tenth the rate of production by CLs. Temporal correlations between the plasma progesterone profile and stages of placental development were also assessed. The rise in plasma progesterone concentrations occurs before differentiation of the chorioallantoic placenta, but progesterone is still high when it degenerates. We conclude that the CLs are the major source of gestational progesterone in N. metallicus. PMID- 11867291 TI - Digestibility and energy value of non-starch polysaccharides in young chickens, ducks and geese, fed diets containing high amounts of barley. AB - The purpose of the present investigation is to compare the ability of chickens, ducks and geese to digest and utilise a diet containing a relatively large amount of barley (40%) rich in beta-glucan (18 g kg(-1)) and NSP (137 g kg(-1)) of which 35 g kg(-1) were soluble non-starch polysaccharides (NSP). The diets were offered to the birds (50 chickens, 40 ducks and 30 geese) in the period from hatching to 42 days of age. The digestibility of NSP was measured during the last week of the growth period using chromic oxide as an indigestible marker. Emphasis was on total NSP, soluble and insoluble NSP and their constituent sugar residues (rhamnose, fucose, arabinose, xylose, mannose, galactose, glucose and glucuronic acid). The degradation of NSP to short chain fatty acids (SCFA) was determined in the small intestine, caeca and large intestine. Although significant differences were found between species to the extent of degradation of individual soluble, insoluble and total NSP residues in the small intestine and caeca, the overall apparent digestibility of total NSP was similar (39-42%). On the basis of the digestibility of the NSP sugar residues and the formation of SCFA in the gut, the energy value of NSP was estimated on 2.8, 3.2 and 2.7 kJ g(-1) NSP ingested (P>0.05) in chickens, ducks and geese, respectively. On average, NSP contributed approximately 3.5% of metabolisable energy (ME) in the three poultry species. PMID- 11867292 TI - Feeding and wounding responses in Hydra suggest functional and structural polarization of the tentacle nervous system. AB - The nervous system of Hydra, a freshwater cnidaria, occurs as dispersed, or diffuse, nerve net throughout the animal. It is widely accepted that in a diffuse nervous system an external stimulus is conducted in all directions over the net. Here I report observations that hydra tentacles respond to feeding and wounding stimuli in a unidirectional manner. Upon contact of a tentacle with a brine shrimp larva during feeding, tissue on the proximal side of the point of contact contracted strongly, whereas tissue on the distal side contracted only very weakly. Feeding a tentacle to which a second tentacle was grafted to the proximal end in the reversed orientation showed that unidirectional conduction, once initiated, was blocked by the reversal of polarity, demonstrating that the distal to proximal polarity of tissue is crucial for unidirectional conduction. Unidirectional conduction was obtained also by mechanically pinching the tissue. The response of tentacles devoid of neurons examined was bidirectional, demonstrating that the nervous system is responsible for the unidirectional responses. These observations suggest that polarized property of the nerve net in hydra tentacles is responsible for the unidirectional tentacle contraction. PMID- 11867293 TI - Reproduction response of Chinese mitten-handed crab (Eriocheir sinensis) fed different sources of dietary lipid. AB - The effect of feeding three semi-purified diets containing different lipid sources (anchovy oil, soybean oil and pork lard) on fecundity, hatchability and egg fatty acid composition of Chinese mitten-handed crab (Eriocheir sinensis) broodstock was compared with a fresh clam diet in a 6-month feeding trial. Broodstock crabs fed the diet containing pork lard showed poor fecundity and low hatchability. Crabs fed the diet containing soybean oil showed improved fecundity; however, no significant improvement in hatchability was observed. Broodstock fed the diet containing anchovy oil showed the highest fecundity and egg hatchability. Eggs from broodstock fed anchovy oil as sole dietary lipid had a higher n-3 polyunsaturated fatty acid (PUFA) content (33.3%) compared with those of crabs fed diets with soybean oil (20.1%) and pork lard (16.3%) as lipid sources. The results indicate a close correlation between: (1) the 20:5n-3 content of the egg lipid and fecundity; (2) the 22:6n-3 content and hatchability; and (3) fecundity, hatchability and n-3/n-6 fatty acid ratio. The results also suggest that each of these n-3 HUFAs may play different and specific roles in crab reproduction and that either must be adequate in the broodstock diet. PMID- 11867294 TI - Energy expenditure and blood flows in thermoregulatory organs during microgravity simulation in rat. Emphasis on the importance of the control group. AB - Rat tail suspension is commonly used to mimic human physiology in space. However, energy metabolism adaptation and related autonomic responses are unknown. To give new insights in energy homeostasis, we determined total energy expenditure (TEE) and blood flow redistribution in thermoregulatory organs during suspension using two control groups of animals widely accepted in the literature: the individually housed (isolated) and restraint rats (horizontally attached to the suspension device). Rats (n=33) were randomly assigned during 14-days to three experimental groups: isolated, suspended, or attached. TEE was assessed by a doubly labeled water method throughout the 14 days, and regional blood flow by radiolabeled microsphere procedure at the end of the protocol. Attachment vs. suspension resulted in a significant decrease in TEE (25%), skin (54%), adrenal (55%) and kidney (42%) blood flows, cardiac index (33%), and plasma corticosterone (50%), whereas total peripheral resistances increased (50%). Isolation vs. attachment triggered an inverse response, of similar amplitude, for all above variables. By comparing isolation and suspension, no overall effect was observed. The striking conclusion of this study is that no clear conclusion can be drawn. The choice of the isolated or attached animals as control profoundly influences the outcome results regarding the effects of simulated weightlessness. Further studies are needed but we favor the attached group as the true control since, from a theoretical point of view, a suspended rat is attached plus suspended. In such conditions, TEE decreases to the same extent in rat and humans during simulated microgravity. When reviewing published experiments, we recommend special attention to the control group used rather than on the effects of suspension as compared to an undefined control. PMID- 11867295 TI - The serostatus approach to HIV prevention and care: cautions and caveats. PMID- 11867296 TI - An analysis of the serostatus approach to fighting the HIV epidemic. PMID- 11867299 TI - Clinical assessment software application (CASA) and immunization coverage rates. PMID- 11867301 TI - The UN General Assembly Special Session on HIV/AIDS: were some lessons of the last 20 years ignored? PMID- 11867302 TI - Ensuring timely access to care for people with HIV infection: a public health imperative. PMID- 11867303 TI - HIV/AIDS stigma: an impediment to public health. PMID- 11867304 TI - The global HIV/AIDS pandemic, structural inequalities, and the politics of international health. AB - In spite of recent advances in treatment and care available in most developed countries, the HIV/AIDS pandemic continues to spread throughout the developing world. Structural inequalities continue to fuel the epidemic in all societies, and HIV infection has increasingly been concentrated in the poorest, most marginalized sectors of society in all countries. The relationship between HIV/AIDS and social and economic development has therefore become a central point in policy discussions about the most effective responses to the epidemic. Important progress has been made in recent United Nations initiatives. Maintaining long-term commitment to initiatives such as the Global Fund to Fight AIDS, Tuberculosis and Malaria is especially important in the wake of September 11 and ensuing events, which threaten to redirect necessary resources to seemingly more urgent security concerns. PMID- 11867305 TI - Ensuring a public health impact of programs to reduce HIV transmission from mothers to infants: the place of voluntary counseling and testing. AB - Since 1999, many African governments have launched programs to offer short-course antiretroviral drug regimens to reduce mother-to child transmission of HIV. HIV testing in prenatal care is the gateway to these antiretroviral regimens. Pilot projects in Africa show an uptake of antiretroviral drugs in 8% to 50% of pregnant women presumed to be HIV infected; often, a minority of eligible women in care received these regimens. Use of lay counselors and rapid onsite HIV testing may alleviate health service barriers. Community education to promote voluntary counseling and testing, which involves men, is the long-term solution. In the short term, possibilities to enhance delivery of an effective intervention include group pretest counseling, universal offer of testing with women having the right to "opt out," universal treatment (mass treatment for those whose HIV status is not determined by voluntary counseling and testing), universal testing with women having the right to "opt out" of learning their test results, and mass treatment for all without testing. PMID- 11867306 TI - Corporate speech and the Constitution: the deregulation of tobacco advertising. AB - In a series of recent cases, the Supreme Court has given businesses powerful new First Amendment rights to advertise hazardous products. Most recently, in Lorillard Tobacco Co v Reilly (121 SCt 2404 [2001]), the court invalidated Massachusetts regulations intended to reduce underage smoking. The future prospects for commercial speech regulation appear dim, but the reasoning in commercial speech cases is supported by only a plurality of the court. A different First Amendment theory should recognize the importance of population health and the low value of corporate speech. In particular, a future court should consider the low informational value of tobacco advertising, the availability of alternative channels of communication, the unlawful practice of targeting minors, and the magnitude of the social harms. PMID- 11867308 TI - Expressive freedom and tobacco advertising: a Canadian perspective. AB - In 1989, Canada enacted the Tobacco Products Control Act (TPCA), which prohibited tobacco advertising, required health warnings on tobacco packaging, and restricted promotional activities. Canada's tobacco companies challenged the TPCA's constitutionality, arguing that it infringed on freedom of expression. Although it seemed likely that the Canadian Supreme Court would uphold the legislation, in 1995 the court declared the impugned provisions to be unconstitutional. The decision is testimony to the constraining force of liberalism on tobacco regulation, but it is also evidence of the power of political will. While the Canadian government could have used the decision to justify withdrawing from further confrontations with powerful commercial interests, it chose instead to enact new tobacco control legislation in 1997. PMID- 11867307 TI - Tobacco, commercial speech, and libertarian values: the end of the line for restrictions on advertising? AB - In June of 2001, the Supreme Court overturned a set of antitobacco measures adopted by the state of Massachusetts designed to protect young people from advertising. Once again, the court expressed its hostility toward measures designed to restrict commercial speech in the name of the social good. In so doing, the court underscored the enduring tension between the libertarian and social welfare dimensions of contemporary democracy and placed into relief the divisions within the American liberal tradition. PMID- 11867309 TI - Medical education for women, 1870. PMID- 11867310 TI - How common is choosing to discontinue treatment for HIV? PMID- 11867311 TI - Effect of nevirapine toxicity on choice of perinatal HIV prevention strategies. AB - OBJECTIVES: This study evaluated the validity of concerns about the toxicity of nevirapine (NVP) that have delayed its implementation as a perinatal HIV prevention strategy. METHODS: A decision analysis model compared 3 strategies: single-dose NVP, short-course zidovudine (ZDV), and no intervention. RESULTS: NVP would prevent more deaths than ZDV and no intervention as long as the rate of NVP toxicity did not exceed, respectively, 9 times that observed in the earlier NVP clinical trial and 42 times that observed in the clinical trial. NVP would be economically preferable to ZDV as long as the rate of toxicity did not exceed 22 times that observed in the clinical trial. CONCLUSIONS: Field implementation of NVP should not be delayed by concerns about its toxicity. PMID- 11867312 TI - Violence during pregnancy among women with or at risk for HIV infection. AB - OBJECTIVES: This study estimated the prevalence of violence during pregnancy in relation to HIV infection. METHODS: Violence, current partnerships, and HIV risk behaviors were assessed among 336 HIV-seropositive and 298 HIV-seronegative at risk pregnant women. RESULTS: Overall, 8.9% of women experienced recent violence; 21.5% currently had abusive partners. Violence was experienced by women in all partnership categories (range = 3.8% with nonabusive partners to 53.6% with physically abusive partners). Neither experiencing violence nor having an abusive partner differed by serostatus. Receiving an HIV diagnosis prenatally did not increase risk. Disclosure-related violence occurred, but was rare. CONCLUSIONS: Many HIV-infected pregnant women experience violence, but it is not typically attributable to their serostatus. Prenatal services should incorporate screening and counseling for all women at risk for violence. PMID- 11867313 TI - HIV-related stigma and knowledge in the United States: prevalence and trends, 1991-1999. AB - OBJECTIVES: This study assessed the prevalence of AIDS stigma and misinformation about HIV transmission in 1997 and 1999 and examined trends in stigma in the United States during the 1990s. METHODS: Telephone surveys with national probability samples of English-speaking adults were conducted in the period 1996 to 1997 (n = 1309) and in 1998 to 1999 (n = 669). Findings were compared with results from a similar 1991 survey. RESULTS: Overt expressions of stigma declined throughout the 1990s, with support for its most extreme and coercive forms (e.g., quarantine) at very low levels by 1999. However, inaccurate beliefs about the risks posed by casual social contact increased, as did the belief that people with AIDS (PWAs) deserve their illness. In 1999, approximately one third of respondents expressed discomfort and negative feelings toward PWAs. CONCLUSION: Although support for extremely punitive policies toward PWAs has declined, AIDS remains a stigmatized condition in the United States. The persistence of discomfort with PWAs, blame directed at PWAs for their condition, and misapprehensions about casual social contact are cause for continuing concern and should be addressed in HIV prevention and education programs. PMID- 11867314 TI - Relationships of stigma and shame to gonorrhea and HIV screening. AB - OBJECTIVES: The purpose of this study was to assess the relationships between stigma and shame associated with seeking treatment for sexually transmitted diseases (STDs) and undergoing testing for gonorrhea and HIV. METHODS: Participants were 847 males and 1126 females (mean age: 24.9 years) in 7 cities. Two scales assessed STD-related stigma and STD-related shame. RESULTS: Rates of stigma and shame were higher among participants without a gonorrhea test in the past year and among those without an HIV test. Sex, age, health service use, previous suspicion of gonorrhea, and low levels of stigma were independently associated with gonorrhea testing. Age, enrollment site, use of health services, gonorrhea testing, and low levels of stigma were independently associated with HIV testing. CONCLUSIONS: Shame is part of the experience of seeking STD-related care, but stigma may be a more powerful barrier to obtaining such care. PMID- 11867315 TI - Changes in HIV seroprevalence and related behaviors among male injection drug users who do and do not have sex with men: New York City, 1990-1999. AB - OBJECTIVES: This study examined HIV prevalence and risk behaviors among male injection drug users (IDUs) who have sex with men and among other male IDUs. METHODS: Male IDUs were interviewed and tested for HIV at a detoxification clinic during 1990 to 1994 and 1995 to 1999. Analyses compared male IDUs who do and do not have sex with men within and between periods. RESULTS: Initially, HIV seroprevalence and risk behaviors were higher among IDUs who have sex with men. Seroprevalence (initially 60.5% vs 48.3%) declined approximately 15% in both groups, remaining higher among those who have sex with men. Generally, injection prevalence, but not sexual risk behaviors, declined. CONCLUSIONS: Male IDUs who have sex with men are more likely to engage in higher-risk behaviors and to be HIV infected. Improved intervention approaches for male IDUs who have sex with men are needed. PMID- 11867316 TI - Age-specific seroprevalence of HIV, hepatitis B virus, and hepatitis C virus infection among injection drug users admitted to drug treatment in 6 US cities. AB - OBJECTIVES: This study measured age-specific seroprevalence of HIV, hepatitis B virus, and hepatitis C virus (HCV) infection among injection drug users (IDUs) admitted to drug treatment programs in 6 US cities. METHODS: Remnant sera collected from persons entering treatment with a history of illicit drug injection were tested for antibodies to HIV, hepatitis C (anti-HCV), and hepatitis B core antigen (anti-HBc). RESULTS: Prevalence of anti-HBc and anti-HCV increased with age and reached 80% to 100% among older IDUs in all 6 cities. Although overall age-specific HIV prevalence was lower than anti-HCV or anti-HBc, this prevalence was greater in the Northeast than in the Midwest and West. CONCLUSIONS: The need continues for effective primary prevention programs among IDUs specifically targeting young persons who have recently started to inject drugs. PMID- 11867318 TI - Analysis of a population-based Pneumocystis carinii pneumonia index as an outcome measure of access and quality of care for the treatment of HIV disease. AB - OBJECTIVES: A population-based Pneumocystis carinii pneumonia (PCP) Index was developed in New York City to identify geographic areas and subpopulations at increased risk for PCP. METHODS: A zip code-level PCP Index was created from AIDS surveillance and hospital discharge records and defined as (number of PCP-related hospitalizations)/(number of persons living with AIDS). RESULTS: In 1997, there were 2262 hospitalizations for PCP among 39 740 persons living with AIDS in New York City (PCP Index =.05691). PCP Index values varied widely across neighborhoods with high AIDS prevalence (West Village =.02532 vs Central Harlem =.08696). Some neighborhoods with moderate AIDS prevalence had strikingly high rates (Staten Island =.14035; northern Manhattan =.08756). CONCLUSIONS: The PCP Index highlights communities in particular need of public health interventions to improve HIV-related service delivery. PMID- 11867317 TI - Impact of highly active antiretroviral treatment on HIV seroincidence among men who have sex with men: San Francisco. AB - OBJECTIVES: This study assessed the countervailing effects on HIV incidence of highly active antiretroviral treatment (HAART) among San Francisco men who have sex with men (MSM). METHODS: Behavioral risk was determined on the basis of responses to cross-sectional community interviews. HIV incidence was assessed through application of an enzyme-linked immunoassay testing strategy. RESULTS: Use of HAART among MSM living with AIDS increased from 4% in 1995 to 54% in 1999. The percentage of MSM who reported both unprotected anal intercourse and multiple sexual partners increased from 24% in 1994 to 45% in 1999. The annual HIV incidence rate increased from 2.1% in 1996 to 4.2% in 1999 among MSM who sought anonymous HIV testing, and the rate was high (5.3%) but stable in a blinded survey of MSM seeking sexually transmitted disease services. CONCLUSIONS: Any decrease in per contact risk of HIV transmission due to HAART use appears to have been counterbalanced or overwhelmed by increases in the number of unsafe sexual episodes. PMID- 11867319 TI - Improving the health of infants on Medicaid by collocating special supplemental nutrition clinics with managed care provider sites. AB - OBJECTIVES: This study tested whether collocation of Special Supplemental Nutrition Program for Women, Infants, and Children (WIC) clinics at managed care provider sites improved health care for infants enrolled in Medicaid and WIC. METHODS: Weights and immunization rates were studied for the 1997 birth cohort of African American infants enrolled in WIC and Medicaid in Detroit, Mich. Infants using traditional WIC clinics and health services were compared with those enrolled under Medicaid in 2 managed care organizations (MCOs), of whom about half obtained WIC services at MCO provider sites. RESULTS: Compared with other infants, those who used collocated WIC sites either were closer to their age appropriate weight or had higher immunization rates when recertified by WIC after their first birthday. Specific benefits (weight gain or immunizations) varied according to the priorities at the collocated sites operated by the 2 MCOs. CONCLUSIONS: Collocation of WIC clinics at MCO sites can improve health care of low-income infants. However specific procedures for cooperation between WIC staff and other MCO staff are required to achieve this benefit. PMID- 11867320 TI - No care for the caregivers: declining health insurance coverage for health care personnel and their children, 1988-1998. AB - OBJECTIVES: This study examined trends in health insurance coverage for health care workers and their children between 1988 and 1998. METHODS: We analyzed data from the annual March supplements of the Current Population Survey (CPS), a Census Bureau survey that collects information about health insurance from a nationally representative sample of noninstitutionalized US residents. RESULTS: Of the health care personnel younger than 65 years, 1.36 million (90% confidence interval [CI] = 1.28 million, 1.45 million) were uninsured in 1998, up 83.4% from 1988; the proportion uninsured rose from 8.4% (90% CI = 7.8%, 9.1%) to 12.2% (90% CI = 11.5%, 12.9%). Declining coverage rates in the growing private-sector health care workforce---and declining health employment in the public sector, which provided health insurance benefits to more of its workers---accounted for the increases. Households with a health care worker included 1.12 million (90% CI = 1.05 million, 1.20 million) uninsured children, accounting for 10.1% (90% CI = 9.5%, 10.8%) of all uninsured children in the United States. CONCLUSIONS: Health care personnel are losing health insurance coverage more rapidly than are other workers. Increasingly, the health care sector is consigning its own workers and their children to the ranks of the uninsured. PMID- 11867321 TI - When the caregiver needs care: the plight of vulnerable caregivers. AB - OBJECTIVES: This study examined the characteristics, activities, and challenges of high-risk informal caregivers. METHODS: Telephone interviews were conducted with a nationally representative cross-section of 1002 informal caregivers. Vulnerable caregivers with poor health or a serious health condition were compared with nonvulnerable caregivers. RESULTS: Thirty-six percent of caregivers were vulnerable. Compared with nonvulnerable caregivers, vulnerable caregivers were more likely to have difficulty providing care, to provide higher-intensity care, to report that their physical health had suffered since becoming a caregiver, to be aged 65 years or older, to be married, and to have less than 12 years of education. CONCLUSIONS: Reliance on informal caregivers without considering the caregiver's ability to provide care can create a stressful and potentially unsafe environment for the caregiver and the care recipient. PMID- 11867322 TI - Smooth moves: bar and nightclub tobacco promotions that target young adults. AB - OBJECTIVES: This article describes the tobacco industry's use of bars and nightclubs to encourage smoking among young adults. METHODS: Previously secret tobacco industry marketing documents were analyzed. RESULTS: Tobacco industry bar and nightclub promotions in the 1980s and 1990s included aggressive advertising, tobacco brand--sponsored activities, and distribution of samples. Financial incentives for club owners and staff were used to encourage smoking through peer influence. Increased use of these strategies occurred concurrently with an increase in smoking among persons aged 18 through 24 years. CONCLUSIONS: The tobacco industry's bar and nightclub promotions are not yet politically controversial and are not regulated by the 1998 Master Settlement Agreement between the industry and the states. Tobacco control advocates should include young adults in research and advocacy efforts and should design interventions to counter this industry strategy to solidify smoking patterns and recruit young adult smokers. PMID- 11867323 TI - Spatial variability in toxicity indicators used to rank chemical risks. AB - OBJECTIVES: This study used 6 different measures of toxicity to explore spatial and statistical variations in relative risk indicators of Toxic Release Inventory emissions. METHODS: Statistical and spatial correlations between the 6 indices were computed for individual South Carolina facilities. RESULTS: Although the 6 toxicity indices are not highly correlated in theory, they have more commonality in practice. There was significant spatial variation in the indices by individual facility level. CONCLUSIONS: Environmental justice researchers must be cognizant of differences in toxicity indices because the choice of the toxicity measure can alter (statistically and spatially) the results of equity analyses and lead to erroneous conclusions. PMID- 11867324 TI - Timing of insurance coverage and use of prenatal care among low-income women. AB - OBJECTIVES: This study examined the relationship between timing of insurance coverage and prenatal care among low-income women. METHODS: Timeliness of prenatal care initiation and adequacy of number of visits were studied among 5455 low-income participants in a larger cross-sectional statewide survey of postpartum women in California during 1994-1995. RESULTS: Although only 2% of women remained uninsured throughout pregnancy, one fifth lacked coverage during the first trimester. Rates of untimely care were highest (> or =64%) among women who were uninsured throughout their pregnancy or whose coverage began after the first trimester; rates were lowest (about 10%) among women who obtained coverage during the first trimester. Women who first obtained Medi-Cal coverage during pregnancy were at low risk of having too few visits. CONCLUSIONS: Timing of prenatal coverage should be considered in research on the relationship between coverage and care use among low-income women. Earlier studies that relied solely on principal payer information, without data on when coverage began, may have led to inaccurate inferences about lack of coverage as a barrier to prenatal care. PMID- 11867325 TI - Neighborhood context and youth cardiovascular health behaviors. AB - OBJECTIVES: This study sought to determine the relationships between race/ethnicity, socioeconomic status (SES), and cardiovascular health behaviors among youths and whether neighborhood characteristics are associated with such behaviors independently of individual characteristics. METHODS: Linear models determined the effects of individual and neighborhood characteristics (SES, social disorganization, racial/ethnic minority concentration, urbanization) on dietary habits, physical activity, and smoking among 8165 youths aged 12 to 21 years. RESULTS: Low SES was associated with poorer dietary habits, less physical activity, and higher odds of smoking. After adjustment for SES, Black race was associated with poorer dietary habits and lower odds of smoking. Hispanic ethnicity was associated with healthier dietary habits, lower levels of physical activity, and lower odds of smoking than non-Hispanic ethnicity. Low neighborhood SES and high neighborhood social disorganization were independently associated with poorer dietary habits, while high neighborhood Hispanic concentration and urbanicity were associated with healthier dietary habits. Neighborhood characteristics were not associated with physical activity or smoking. CONCLUSIONS: Changes in neighborhood social structures and policies that reduce social inequalities may enhance cardiovascular health behaviors. PMID- 11867327 TI - Effect of racial/ethnic misclassification of American Indians and Alaskan Natives on Washington State death certificates, 1989-1997. AB - OBJECTIVES: This study examined effects of racial/ethnic misclassification of American Indians and Alaskan Natives on Washington State death certificates. METHODS: Probabilistic record linkage were used to match the 1989-1997 state death files to the Northwest Tribal Registry. RESULTS: We identified matches for 2819 decedents, including 414 (14.7%) who had been misclassified as non-American Indians and Alaskan Natives on the death certificates. The likelihood of being correctly classified increased 3-fold for each higher level of American Indian and Alaskan Native ancestry (odds ratio = 2.88; 95% confidence interval [CI] = 2.51, 3.30) and decreased by 6.9% per calendar year (95% CI = 2.0, 11.5). CONCLUSIONS: Systematic biases on death certificates in Washington State persist. Methods to reduce misclassification can improve data quality and enhance efforts to measure and reduce racial/ethnic health disparities. PMID- 11867326 TI - Effect on smoking cessation of switching nicotine replacement therapy to over-the counter status. AB - OBJECTIVES: This study examined whether the change in nicotine replacement therapy sales from prescription to over the counter (OTC) status affected smoking cessation. METHODS: We used the 1993-1999 Massachusetts Tobacco Surveys to compare data from adult current smokers and recent quitters before and after the OTC switch. RESULTS: No significant change over time occurred in the proportion of smokers who used nicotine replacement therapy at a quit attempt in the past year (20.1% pre-OTC vs 21.4% post-OTC), made a quit attempt in the past year (48.1% vs 45.2%), or quit smoking in the past year (8.1% vs 11.1%). Fewer non Whites used nicotine replacement therapy after the switch (20.7% pre-OTC vs 3.2% post-OTC, P =.002), but the proportion of Whites using nicotine replacement therapy did not change significantly (20.6% vs 24.0%). CONCLUSIONS: We observed no increase in Massachusetts smokers' rates of using nicotine replacement therapy, making a quit attempt, or stopping smoking after nicotine replacement therapy became available for OTC sale. There appear to be other barriers to the use of nicotine replacement therapy besides visiting a physician, especially among minority smokers. PMID- 11867328 TI - Subgroups of refusers in a disability prevention trial in older adults: baseline and follow-up analysis. AB - OBJECTIVES: This study explored differences between refusers and participants in a longitudinal study with extensive baseline and follow-up information. METHODS: Results of a trial comparing 791 participants and 401 community-residing older adults who refused to participate in a study concerning preventive home visits were examined. Information was collected from interviews, insurance records, and government files. RESULTS: Despite similarities in terms of age, sex, and self perceived health at baseline, 3-year follow-up data indicated that refusers had a 1.58-fold higher risk of entering a nursing home than participants. There were additional differences between refusers and participants when refusers were categorized in 4 subgroups based on self-reported reason for refusal (too ill, too healthy, no interest, and other reasons). CONCLUSIONS: Future studies should include follow-up data to allow comparisons between refusers and participants and should address the presence of multiple subgroups of refusers. PMID- 11867330 TI - Distinct roles for lung collectins in pulmonary host defense. PMID- 11867331 TI - Understanding the mechanisms of infant respiratory distress and chronic lung disease. PMID- 11867332 TI - Neutrophil elastase: path clearer, pathogen killer, or just pathologic? PMID- 11867333 TI - m3 muscarinic acetylcholine receptor regulation in the airway. PMID- 11867334 TI - When there is a fungus among us, what makes it virulent? PMID- 11867335 TI - Surfactant protein-A--deficient mice display an exaggerated early inflammatory response to a beta-resistant strain of influenza A virus. AB - Surfactant protein (SP)-A is a member of the collectin family of proteins. In vitro, SP-A binds influenza A virus (IAV), neutralizes infectivity, and enhances uptake by macrophages. SP-D also binds and neutralizes certain strains of IAV. To determine if SP-A has a role in protecting the intact animal against IAV infection, we inoculated gene-targeted SP-A-deficient mice (-/-) and littermate controls (+/+) with either saline or increasing doses of an IAV strain that binds SP-A but not SP-D. IAV was more virulent in SP-A-/- compared with +/+ mice, with a significantly lower mean lethal dose (LD(50)) and significantly greater weight loss during infection. SP-A-/- mice also had increased airway epithelial injury and more alveolar cellular infiltrates than +/+ mice. On Day 2, SP-A-/- mice had more neutrophils and higher MIP-2 levels in the lung than +/+ mice. We conclude the altered host response and increased susceptibility to X-79Delta167 infection in SP-A-/- mice reflects a protective role for SP-A in regulating the host response to IAV. Because the recovery of virus from lung homogenates on Days 2 and 6 after inoculation was comparable in -/- and +/+ mice, we speculate SP-A reduces IAV virulence independently of direct viral neutralization. PMID- 11867336 TI - Pro- and anti-inflammatory cytokines regulate insulin-like growth factor binding protein production by fetal rat lung fibroblasts. AB - The inflammatory response of the lung to noxious factors contributes to the pathogenesis of chronic lung injury. Inflammatory mediators regulate the insulin like growth factor (IGF) system, a key modulator of lung fibroblast proliferation. The activity of IGFs is regulated by IGF-binding proteins (IGFBPs) secreted by lung cells. To investigate the regulation of lung fibroblast IGFBPs by cytokines, we exposed 19-d fetal rat lung fibroblasts to various pro- and anti inflammatory mediators. IGFBP abundance in conditioned medium (CM) was measured by ligand blot and RNA transcript abundance by RNase protection assays. Fetal rat lung fibroblasts exposed to interleukin (IL)-1beta or tumor necrosis factor (TNF) alpha for 48 h demonstrated increased abundance of CM IGFBP-3 (5.9- and 4.7-fold increases for IL-1beta and TNF-alpha, respectively) and IGFBP-4 (5.7- and 7.4 fold increases for IL-1beta and TNF-alpha, respectively) that was accompanied by a small increase in IGFBP-4 mRNA and a larger increase in IGFBP-3 mRNA abundance. IGFBP-4 specific proteolysis was examined in CM collected from fetal rat lung fibroblasts after incubation with serum-free medium (SFM), IL-1beta, or TNF-alpha for 48 h. Cell-free aliquots of SFM-CM incubated at 37C for 24 h showed a 65% decrease in IGFBP-4 abundance that was inhibited by 1,10-phenanthroline. In contrast, CM from cells exposed to IL-1beta or TNF-alpha incubated at 37 degrees C for 24 h did not show a significant decrease in IGFBP-4 abundance unless IGF-I was present during the cell-free incubation. Addition of IGFBP-3 to aliquots of SFM-CM reversed the IGF-I-mediated acceleration of IGFBP-4 proteolysis. Similarly, addition of IGFBP-3 to cells in culture increased the accumulation of CM IGFBP-4. These results demonstrate that cytokines regulate IGFBP production and clearance by fetal lung cells and suggest a mechanism by which cytokines regulate cell proliferation following lung injury. PMID- 11867337 TI - Protection against acute lung injury by intravenous or intratracheal pretreatment with EPI-HNE-4, a new potent neutrophil elastase inhibitor. AB - Excessive accumulation of active neutrophil elastase (NE) in pulmonary fluids and tissues of patients with cystic fibrosis (CF) is thought to act on the lungs, compromising their structure and function. The aim of this study was to investigate the in vitro and in vivo protective effect of a new, rapidly acting, potent (Ki = 5.45 x 10(-12) M and Kon = 8 x 10(6) M(-1) s(-1)) and specific human NE inhibitor, EPI-HNE-4, engineered from the Kunitz domain. The results demonstrated that this inhibitor was able to (i) effectively inhibit in vitro the high levels of active NE present in a medium as complex as sputum from children with CF, with a measured IC(50) equal or close to the calculated IC(50) in 60% of cases, and (ii) almost completely block (91%) the N-formyl-methionine-leucine phenylalanine-induced migration of purified human neutrophils across a Matrigel basement membrane. Intratracheal administration (250, 175, or 100 microg per rat) of the inhibitor 5 min before instillation of pure human NE (HNE) (150 microg per rat) to rats induced effective, dose-dependent protection of the lungs, 4 h later, from hemorrhage, serum albumin leakage, residual active NE, and discrete neutrophil influx in air spaces induced by instillation of pure HNE. Intravenous administration (3 mg per rat) of EPI-HNE-4, 15 min before instillation of the soluble fraction of pooled sputum (delivering 120 microg of active NE per rat) from children with CF, effectively reduced (64%), 4 h later, the massive neutrophil influx induced by sputum instillation. Overall, these data strongly suggest that associated aerosol and systemic administration of EPI-HNE-4 would be beneficial in the treatment of CF. PMID- 11867338 TI - Structure and transcription of the human m3 muscarinic receptor gene. AB - We have isolated and characterized the human m3 muscarinic receptor gene and its promoter. Using 5' rapid amplification of cDNA ends (RACE), internal polymerase chain reaction (PCR), and homology searching to identify EST clones, we determined that the cDNA encoding the m3 receptor comprises 4,559 bp in 8 exons, which are alternatively spliced to exclude exons 2, 4, 6, and/or 7; the receptor coding sequence occurs within exon 8. Analysis of P1 artificial chromosome (PAC) and bacterial artificial chromosome (BAC) clones and of PCR- amplified genomic DNA, and homology searching of human chromosome 1 sequence provided from the Sanger Centre (Hinxton, Cambridge, UK) revealed that the m3 muscarinic receptor gene spans at least 285 kb. A promoter fragment containing bp -1240 to +101 (relative to the most 5' transcription start site) exhibited considerable transcriptional activity during transient transfection in cultured subconfluent, serum-fed canine tracheal myocytes, and 5' deletion analysis of promoter function revealed the presence of positive transcriptional regulatory elements between bp 526 and -269. Sequence analysis disclosed three potential AP-2 binding sites in this region; five more AP-2 consensus binding motifs occur between bp -269 and +101. Cotransfection with a plasmid expressing human AP-2alpha substantially increased transcription from m3 receptor promoter constructs containing 526 or 269 bp of 5' flanking DNA. Furthermore, m3 receptor promoter activity was enhanced by long-term serum deprivation of canine tracheal myocytes, a treatment that is known to increase AP-2 transcription-promoting activity in these cells. Together, these data suggest that expression of the human m3 muscarinic receptor gene is regulated in part by AP-2 in airway smooth muscle. PMID- 11867339 TI - Complementation of a capsule deficient Cryptococcus neoformans with CAP64 restores virulence in a murine lung infection. AB - Cryptococcosis is a systemic infection in humans caused by the opportunistic fungal pathogen, Cryptococcus neoformans. The infection usually presents as chronic meningoencephalitis, but infects via the respiratory tract. A polysaccharide capsule is a major virulence factor, which allows the yeast to resist host defenses. However, the essential role of the capsule in allowing it to resist host defenses during the initial lung infection has not been clearly shown. A mutant acapsular C. neoformans strain 602 was complemented with the CAP64 gene to obtain an encapsulated strain, TYCC38-602. TYCC38-602 persisted in the lungs of C.B-17 mice after intratracheal inoculation and disseminated to the brain, whereas the mutant acapsular 602 and the plasmid control transformant CIP3 602 strains grew less readily in the lung and were infrequently detected in the brain. T cell-mediated immunity, developed to the encapsulated organism, was required to control growth within the lungs and had a significant impact on numbers of yeasts detected in the brain. The parent acapsular strain, but not the transformant control, also required T cells for optimal inhibition of growth within the lung, but not for maintaining control of the colony-forming units (cfu) in the brain. In summary, the cryptococcal capsule plays an important role in lung virulence and dissemination to the brain, and intact immunity is required to control lung growth of the encapsulated yeast. PMID- 11867340 TI - Effect of IgA on respiratory burst and cytokine release by human alveolar macrophages: role of ERK1/2 mitogen-activated protein kinases and NF-kappaB. AB - Human alveolar macrophages (HAM) express FcalphaR receptors for immunoglobulin (Ig)A which could link humoral and cellular branches of lung immunity. Here, we investigate the effects of polymeric (p-IgA) and secretory (S-IgA) IgA interaction with Fc(alpha)R on lipopolysaccharide (LPS)- and phorbol myristate acetate (PMA)-activated respiratory burst and TNF-alpha release by HAM. Activation of HAM with LPS and PMA increases the respiratory burst and TNF-alpha release through activation of the extracellular signal-related protein kinases 1 and 2 (ERK1/2) pathway, because these effects are inhibited by treatment of HAM with PD98059, a selective inhibitor of mitogen-activated protein (MAP)/ERK kinases (MEK) pathway. S-IgA and p-IgA downregulate the LPS-increased respiratory burst in HAM through an inhibition of ERK1/2 activity. In contrast, p- and S-IgA induce an increase in the respiratory burst of PMA-treated HAM. This effect is associated with an upregulation by IgA of the PMA-induced phosphorylation of ERK1/2 and is also inhibited by PD98059. Moreover, p-IgA and S-IgA enhance TNF alpha release by HAM through an alternative pathway distinct from ERK1/2. Because LPS is known to activate nuclear factor-kappaB (NF-kappaB) in HAM, we evaluate the effect of IgA on NF-kappaB. Treatment of HAM with LPS, p- and S-IgA, but not PMA, induces NF-kappaB activation through IkappaBalpha phosphorylation and subsequent proteolysis. Antioxidants, namely N-acetylcysteine (NAC) and glutathione (GSH), have no effects on IgA-mediated NF-kappaB nuclear translocation and only a minor and late effect on that of LPS, suggesting that reactive oxygen intermediates (ROI) play a minor role in HAM activation through NF-kappaB. TNF-alpha release by LPS-activated HAM is sensitive to NF-kappaB inhibition and only partly to oxidant scavenging. In contrast, TNF-alpha release by IgA-treated HAM is not dependent on oxidants and only partly dependent on NF kappaB. Our results show a differential HAM regulation by IgA through both dependent and independent modulation of ERK pathway. In addition, IgA activates NF-kappaB and this effect was independent on oxidants. These data may help to understand the role of IgA in both lung protection and inflammation. PMID- 11867341 TI - Origin, differentiation, and maturation of human pulmonary veins. AB - Recent studies on human embryonic and fetal lungs show that the pulmonary arteries form by vasculogenesis. Little is known of the early development of the pulmonary veins. Using immunohistochemical techniques and serial reconstruction, we studied 18 fetal and neonatal lungs. Sections were stained with antibodies specific for endothelium (CD31, von Willebrand factor) and smooth muscle (alpha and gamma smooth muscle actin, smooth muscle myosin, calponin, caldesmon, and desmin) and antibodies specific for the matrix glycoprotein tenascin, the receptor protein tyrosine kinase EphB4, and its ligand ephrinB2. Kiel University raised antibody number 67 (Ki67) expression allowed qualitative assessment of cell replication. By 34 d gestation, there was continuity between the aortic sac, pulmonary arteries, capillaries, pulmonary veins, and atrium. The pulmonary veins formed by vasculogenesis in the mesenchyme surrounding the terminal buds during the pseudoglandular period and probably by angiogenesis in the canalicular and alveolar stages. EphB4 and ephrinB2 did not distinguish between presumptive venous and arterial endothelium as they do in mouse. All venous smooth muscle cells derived directly from the mesenchyme, gradually acquiring smooth muscle specific proteins from 56 d gestation. Thus, both pulmonary arteries and veins arise by vasculogenesis, but the origins of their smooth muscle cells and their cytoskeletal protein content are different. PMID- 11867342 TI - Modulation of cadherin and catenins expression by tumor necrosis factor-alpha and dexamethasone in human bronchial epithelial cells. AB - Asthma is an inflammatory disease, and the epithelial mesenchymal unit appears to be of importance in regulating the disease mechanisms. Cell-cell adhesion plays an important role in tissue morphogenesis and homeostasis and is commonly mediated by cadherins, a family of Ca(2+)-dependent transmembrane adhesion receptors. The cadherin family is involved in control of the cellular architecture. Proinflammatory cytokines such as tumor necrosis factor (TNF)-alpha are involved in asthma and may interfere with epithelial integrity. In the present study, we investigated the role of TNF-alpha and dexamethasone on the expression of E-cadherin, beta-catenin, and gamma-catenin. We used two bronchial epithelial cell models: primary small airway epithelial cell cultures and primary culture obtained from human bronchial tubes. After 48 h of TNF-alpha stimulation with or without dexamethasone expression of E-cadherin, beta-catenin and gamma catenin were analyzed using Western blot analysis and immunofluorescence. This study showed a decrease in the expression of adhesion molecules in both epithelial cell cultures after stimulation. Dexamethasone and anti-TNF-alpha inhibited this effect. In unstimulated cells, E-cadherin and beta- and gamma catenin expression was membranous, expressed only on the lateral cell wall with minimal cytoplasmic expression. Immunoreactivity was cytoplasmic in stimulated cells. We demonstrated, using Western blot analysis and immunofluorescence, that proinflammatory cytokines could be responsible for structural damage to the epithelium and that this process was potentially reversed by steroids. PMID- 11867343 TI - L-Arginine uptake and metabolism following in vivo silica exposure in rat lungs. AB - Pulmonary inflammation increases nitric oxide (NO) production via inducible nitric oxide synthase (iNOS). This study was performed to determine some of the factors that affect the availability of the NOS substrate, L-arginine (L-arg), in the intact lung subjected to silica-induced inflammation. Nitrate production, as an index of NO production, was significantly greater in silica-exposed lungs (53.5 +/- 12.1 nmol/90 min) compared with controls (22.5 +/- 5.1 nmol/90 min, P < 0.05). This was accompanied by greater (P < 0.0001) 90-min [(3)H]L-arg uptake (62 +/- 3% control, 82 +/- 1% silica), a significantly (P < 0.005) increased permeability-surface area product for L-arg (0.28 +/- 0.05 ml/min control, 0.63 +/- 0.07 ml/min silica), and a significantly (P < 0.001) increased urea production (1.16 +/- 0.08 micromol/90 min control, 1.77 +/- 0.06 micromol/90 min silica). There was no difference in eNOS protein between groups and eNOS mRNA was not detectable in either group, whereas silica exposure resulted in the appearance of both iNOS protein and mRNA. Silica exposure increased CAT-1 and CAT 2 mRNA approximately 8-fold compared with controls. We conclude that the increase in NO production in silica-exposed lungs was associated with increased L-arg uptake from the vasculature, presumably resulting from increased CAT-1 and CAT-2, and by increased L-arg metabolism via arginase. PMID- 11867344 TI - Elastolysis by proteinase 3 and its inhibition by alpha(1)-proteinase inhibitor: a mechanism for the incomplete inhibition of ongoing elastolysis. AB - An excess of proteinase 3 (Pr3) is an assumed risk factor for elastin loss in chronic obstructive pulmonary disease. This study compared the degradation of [(14)C]elastin by Pr3 and its inhibition by alpha(1)-proteinase inhibitor (alpha(1)-PI) with the analogous reactions involving two other neutrophil serine proteases, human leukocyte elastase (HLE) and cathepsin G (CatG). The elastolytic rate catalyzed by Pr3 was estimated to be half of that of CatG and one-eighth of that of HLE. Evidence was obtained that indicated that absorption of Pr3 by the substrate was much less than that of HLE or CatG, and that the majority of absorbed Pr3 was highly mobile. These properties are consistent with the observation that elastolysis by Pr3 was almost completely and stoichiometrically inhibited by alpha(1)-PI even under conditions in which the protease had been preincubated with the substrate. In contrast, alpha(1)-PI in large molar excess was unable to inhibit completely ongoing elastolysis of the same substrate by HLE or CatG. An interfacial nonisotropic reaction mechanism has been proposed to address the incomplete inhibition of ongoing elastolysis. Pr3 was identified as being the most abundant neutrophil serine protease. However, two findings reported here, namely the low rate of elastolysis by Pr3 and the high efficacy of alpha(1)-PI against ongoing elastolysis by Pr3, imply that Pr3 might not necessarily be a major contributor to neutrophil-mediated elastin loss. PMID- 11867345 TI - Isolation and expression of the human hPF20 gene orthologous to Chlamydomonas PF20: evaluation as a candidate for axonemal defects of respiratory cilia and sperm flagella. AB - Primary ciliary dyskinesia (PCD) is a heterogeneous congenital disorder characterized by bronchiectasis and chronic sinusitis, sometimes associated with situs inversus (i.e., Kartagener's syndrome) and male infertility. At the cell level, the disease phenotype includes various axonemal abnormalities of respiratory cilia and sperm flagella. We have previously isolated DNAI1, the first gene involved in these diseases in patients lacking outer dynein arms. In this study, designed to find additional genes for other axonemal defects, we report the isolation of a novel human gene, hPF20, which is orthologous to Chlamydomonas pf20. The hPF20 gene is expressed as two major transcripts: one is expressed in testis only, whereas the second is weakly expressed in many other tissues. As flagella of Chlamydomonas strains carrying pf20 mutations lack the axonemal central complexes, we tested the involvement of the hPF20 gene in the disease phenotype of five patients in whom cilia or flagella display abnormal central complexes. Five intragenic polymorphisms were identified and used to exclude hPF20 in two consanguineous patients, while no mutation was found in the remaining patients. However, given the genetic heterogeneity of PCD, we consider that this gene remains a good candidate to be investigated in patients with abnormal central complexes. PMID- 11867346 TI - Inward rectifier K(+) current in human bronchial smooth muscle cells: inhibition with antisense oligonucleotides targeted to Kir2.1 mRNA. AB - Inward rectifier K(+) (Kir) channels play an important role in forming membrane potential and then modulating muscle tone in certain types of smooth muscles. In cultured human bronchial smooth muscle cells (hBSMCs), Kir current was identified using whole-cell voltage clamp techniques and explored by using RT-PCR analysis of mRNA, Western blotting, and antisense oligonucleotide methods to block the synthesis of Kir channel protein. The K(+) current with strong inward rectification and high K(+) ion selectivity was observed. The current was unaffected by 4-aminopyridine, glibenclamide, and charybdotoxin, and hardly inhibited by tetraethylammonium, but was potently inhibited by extracellular Ba(2+). The IC(50) value of external Ba(2+) was approximately 1.3 microm. RT-PCR analysis of mRNA showed transcripts for Kir2.1, but not Kir1.1, Kir2.2, or Kir2.3. Treatment of cells with antisense oligonucleotides targeted to Kir2.1 resulted in a decrease in the current density of the Kir current and Kir protein expression, as compared with the mismatch-treated cells, whereas the current density of 4-AP-sensitive K(+) currents (K(V)) remained unaffected. The application of Ba(2+) markedly depolarized the membrane. These results demonstrate that Kir channel is present in human bronchial smooth muscle cells, and the Kir2.1 gene encodes the Kir channel protein in these cells. PMID- 11867347 TI - Snoring as a risk factor for type II diabetes mellitus: a prospective study. AB - To examine the association between snoring and risk of developing type II diabetes mellitus, the authors analyzed data from the Nurses' Health Study cohort. This analysis included 69,852 US female nurses aged 40-65 years without diagnosed diabetes, cardiovascular disease, or cancer at baseline in 1986. Snoring patterns were ascertained by questionnaire. During 10 years of follow-up, 1,957 women were diagnosed with type II diabetes. In analyses adjusted for age and body mass index, snoring was associated with risk of diabetes (for occasional snoring vs. nonsnoring, relative risk (RR) = 1.48 (95% confidence interval (CI): 1.29, 1.70); for regular snoring vs. nonsnoring, RR = 2.25 (95% CI: 1.91, 2.66); p for trend < 0.0001). Further adjustment for other diabetes risk factors and sleeping-related covariates only slightly attenuated the risk (for occasional snoring, RR = 1.41 (95% CI: 1.22, 1.63); for regular snoring, RR = 2.03 (95% CI: 1.71, 2.40); p for trend < 0.0001). Analyses stratified by body mass index, smoking history, or parental history of diabetes showed a consistent association between snoring and diabetes within the categories of these variables. These results suggest that snoring is independently associated with elevated risk of type II diabetes. PMID- 11867348 TI - Invited commentary: to sleep, perchance to discover. PMID- 11867350 TI - Exploratory spatial analysis of pilot fatality rates in general aviation crashes using geographic information systems. AB - Geographic information systems and exploratory spatial analysis were used to describe the geographic characteristics of pilot fatality rates in 1983-1998 general aviation crashes within the continental United States. The authors plotted crash sites on a digital map; rates were computed at regular grid intersections and then interpolated by using geographic information systems. A test for significance was performed by using Monte Carlo simulations. Further analysis compared low-, medium-, and high-rate areas in relation to pilot characteristics, aircraft type, and crash circumstance. Of the 14,051 general aviation crashes studied, 31% were fatal. Seventy-four geographic areas were categorized as having low fatality rates and 53 as having high fatality rates. High-fatality-rate areas tended to be mountainous, such as the Rocky Mountains and the Appalachian region, whereas low-rate areas were relatively flat, such as the Great Plains. Further analysis comparing low-, medium-, and high-fatality rate areas revealed that crashes in high-fatality-rate areas were more likely than crashes in other areas to have occurred under instrument meteorologic conditions and to involve aircraft fire. This study demonstrates that geographic information systems are a valuable tool for injury prevention and aviation safety research. PMID- 11867351 TI - Effects of Maryland's law banning "Saturday night special" handguns on homicides. AB - Small, inexpensive, often poorly made handguns known as "Saturday night specials" are disproportionately involved in crime. Maryland banned the sale of Saturday night specials effective January 1, 1990. During the 2 years between the law's passage in 1988 and its effective date, legal handgun sales in Maryland were 34% higher than expected (p = 0.09). Interrupted time-series analysis of age-adjusted homicide rates for 1975-1998 with statistical controls for trends in two neighboring states, social and economic variables, and temporal patterns in Maryland's homicide rates was used to assess the effect of the law. Estimates of the Saturday night special ban effects depended on the assumption made about the timing of the law's effects. Models that assumed a delayed or gradual effect of the Saturday night special ban produced estimates indicating that firearm homicide rates were 6.8-11.5% lower than would have been expected without the Saturday night special ban (p < or =0.05). The model that assumed an immediate, constant change in response to the law showed no law effect, unless an outlier was excluded from the analysis. Excluding this outlier, the model estimated a 15% increase in firearm homicides associated with the Saturday night special ban. None of the models revealed significant law effects on nonfirearm homicides. PMID- 11867352 TI - Suicide rates and religious commitment in young adult males in Utah. AB - Previous studies have used population data to demonstrate an inverse association between suicide rates and religious commitment. This report examines Utah suicide rates for young men aged 15-34 years, stratified by their membership in and commitment to the Church of Jesus Christ of Latter-day Saints (LDS), the predominant religion in Utah. All state death records for males from 1991 to 1995 were obtained and linked to LDS church deceased membership records to obtain a measure of religious commitment that is not self-reported. Religious commitment for LDS church members was determined by age-appropriate priesthood office. Of the 27,738 male deaths reported, 15,555 (56%) linked to an LDS church record using a probabilistic linking program. Using active (high religious commitment) LDS as the reference group, the less-active (low religious commitment) LDS group had relative risks of suicide ranging from 3.28 (ages 15-19 years) to 7.64 (ages 25-29 years); nonmembers of the LDS church had relative risks ranging from 3.43 (ages 15-19 years) to 6.27 (ages 20-24 years). Although the mechanism of the association is unclear, higher levels of religiosity appear to be inversely associated with suicide. PMID- 11867353 TI - Impact of breastfeeding on the mobilization of lead from bone. AB - To evaluate the hypothesis that lactation stimulates lead release from bone to blood, the authors analyzed breastfeeding patterns and bone lead concentrations as determinants of blood lead levels among 425 lactating women in Mexico City for 7 months after delivery (1994-1995). The authors measured in vivo patella and tibia lead concentrations at 1 month postpartum using K x-ray fluorescence. Maternal blood samples and questionnaire information were collected at delivery and at 1, 4, and 7 months postpartum. Blood lead was analyzed using graphite furnace atomic absorption spectroscopy. Mean blood lead level at delivery was 8.4 microg/dl (range: 1.8--23.4). Mean cortical and trabecular lead levels were 10.6 microg/g (range: nondetectable to 76.5) and 15.3 microg/g (range: nondetectable to 85.9), respectively, reflecting a population with elevated and diverse past and current lead exposure. The association of bone lead and breastfeeding with blood lead was estimated using generalized estimating equations. Breastfeeding practices and maternal bone lead were important predictors of blood lead level. After adjustment for bone lead and environmental exposure, women who exclusively breastfed their infants had blood lead levels that were increased by 1.4 microg/dl and women who practiced mixed feeding had levels increased by 1.0 microg/dl, in relation to those who had stopped lactation. These results support the hypothesis that lactation is directly related to the amount of lead released from bone. PMID- 11867354 TI - Effect of caffeine exposure during pregnancy on birth weight and gestational age. AB - Epidemiologic studies have been unable to conclusively evaluate whether caffeine intake during pregnancy is associated with reduced birth weight and/or fetal growth restriction. The authors conducted a prospective, population-based cohort study to investigate the effect of caffeine on birth weight, gestational age, and birth weight standardized for gestational age (birth weight ratio). Of 953 women recruited in early pregnancy in Uppsala County, Sweden, from 1996 to 1998, 873 women delivering liveborn singleton infants were included in the analysis. Caffeine exposures were ascertained from in-person interviews at 6-12 and 32-34 completed gestational weeks, and maternal plasma was analyzed for cotinine levels as an indicator of smoking. Analysis of variance was used to estimate the effect of caffeine on birth weight, gestational age at delivery, and birth weight ratio after accounting for the effects of other covariates, such as maternal sociodemographic characteristics, plasma cotinine, and pregnancy symptoms. There were no associations between caffeine consumption and birth weight, gestational age, and birth weight ratio, neither when caffeine exposure was averaged from conception to the 32nd to 34th gestational weeks, nor when caffeine exposure was stratified by trimesters of pregnancy. These results do not support an association between moderate caffeine consumption and reduced birth weight, gestational age, or fetal growth. PMID- 11867356 TI - Residential magnetic fields and the risk of breast cancer. AB - Chronic exposure to 60-Hz magnetic fields may increase the risk of breast cancer by suppressing the normal nocturnal production of melatonin. This population based case-control study investigated whether such exposure is associated with an increased risk of breast cancer in women aged 20-74 years from the greater Seattle, Washington, area. Cases were diagnosed between November 1992 and March 1995 (n = 813); controls were identified by random digit dialing and were frequency matched by 5-year age groups (n = 793). Exposure was estimated using magnetic field measurements in the home at diagnosis, wiring configuration of all homes occupied in the 10 years prior to diagnosis, and self-reported measures of at-home electric appliance use. Odds ratios and 95% confidence intervals were estimated using conditional logistic regression with adjustment for other potential risk factors. Risk did not increase with measured nighttime bedroom magnetic field level, wiring configuration of the home at diagnosis, weighted summary wire codes of all homes occupied 5 and 10 years prior to diagnosis, or reported use of common household appliances, including bed-warming devices. These data do not support the hypothesis that exposure to residential magnetic fields is associated with an increased risk of developing breast cancer. PMID- 11867355 TI - Endogenous postmenopausal hormones and carotid atherosclerosis: a case-control study of the atherosclerosis risk in communities cohort. AB - Studies examining the relation between endogenous postmenopausal hormone levels and cardiovascular disease have yielded conflicting results. After excluding women with a history of hormone replacement therapy (HRT) use, the authors conducted a US case-control study in 1987-1992 comparing endogenous postmenopausal hormone levels in women with and without significant carotid atherosclerosis in the Atherosclerosis Risk in Communities (ARIC) cohort. Atherosclerosis was assessed by using B-mode ultrasound to measure carotid artery intimal-medial thickness (IMT). Cases (n = 182) were postmenopausal women with average IMT measurements greater-than-or-equal the 95th percentile. Controls (n = 182) were frequency matched to cases on age and ARIC center and had IMT measurements < the 75th percentile. After adjustment for cardiovascular risk factors, no association was found between the odds of atherosclerosis and increasing quartiles of estrone, dehydroepiandrosterone sulfate, or androstenedione. Compared with participants in the lowest quartile of sex hormone binding globulin (SHBG), those in the highest quartile had a significantly lower odds of atherosclerosis (odds ratio = 0.48, 95% confidence interval: 0.24, 0.97). Similarly, participants in the highest quartile of total testosterone had a lower odds of atherosclerosis (odds ratio = 0.38, 95% confidence interval: 0.20, 0.74). The authors found higher total testosterone and SHBG to be inversely related to carotid atherosclerosis, suggesting their potential importance in reducing atherosclerotic risk in postmenopausal women not using HRT. PMID- 11867357 TI - Gender, alcohol consumption, and renal cell carcinoma. AB - The nature of the association between alcohol consumption and renal cell carcinoma (RCC) is not well understood, but there are indications of effect modification by gender. The authors report data from a population-based case control study conducted in Iowa from 1986 to 1989. RCC cases (261 men and 145 women) were identified through the Iowa Cancer Registry, while controls (1,598 men and 831 women) were randomly selected from the general population, frequency matched on age and gender. Subjects provided detailed information on a mailed questionnaire regarding demographic, anthropometric, lifestyle, dietary, and medical history risk factors. In age-adjusted analysis, there was a decrease in risk for women who reported consuming more than three servings (median among drinkers) of alcohol per week (odds ratio = 0.5, 95% confidence interval: 0.2, 0.9) compared with never drinkers. No evidence of an association among men was noted (odds ratio = 1.1, 95% confidence interval: 0.8, 1.5). Multivariate adjustment for anthropometric, lifestyle, smoking, and dietary factors did not alter the findings. Analysis by type of alcohol suggested that the inverse association was strongest for beer consumption, but estimates were imprecise. These findings suggest an inverse association of alcohol consumption and RCC development among women but not among men. PMID- 11867358 TI - Lung function in relation to intake of carotenoids and other antioxidant vitamins in a population-based study. AB - Accumulating evidence suggests that dietary antioxidant vitamins are positively associated with lung function. No evidence exists regarding whether dietary carotenoids other than beta-carotene are related to pulmonary function. In 1995- 1998 the authors studied the association of forced expiratory volume in 1 second and forced vital capacity as the percentage of the predicted value (FEV(1)% and FVC%, respectively) after adjustment for height, age, gender, and race with the intakes of several carotenoids (alpha-carotene, beta-carotene, beta cryptoxanthin, lutein/zeaxanthin, and lycopene) in a random sample of 1,616 men and women who were residents of western New York State, aged 35--79 years, and free from respiratory disease. They observed significant associations of lutein/zeaxanthin and vitamins C and E with FEV(1)% and FVC% using multiple linear regression after adjustment for total energy intake, smoking, and other covariates. When they analyzed all of these antioxidant vitamins simultaneously, they observed the strongest association of vitamin E with FEV(1)% and of lutein/zeaxanthin with FVC%. The differences in forced expiratory volume in 1 second and forced vital capacity associated with a decrease of 1 standard deviation of dietary vitamin E or lutein/zeaxanthin were equivalent to the influence of approximately 1--2 years of aging. Their findings support the hypothesis that carotenoids, vitamin C, and vitamin E may play a role in respiratory health and that carotenoids other than beta-carotene may be involved. PMID- 11867359 TI - Use of oral antimicrobials decreases serum enterolactone concentration. AB - The lignan enterolactone, a phytoestrogen, may protect against hormone-dependent cancers and cardiovascular diseases. It is produced by the intestinal microflora from dietary precursors. Because of the pronounced impact of antimicrobials on the intestinal microflora, the authors examined whether serum enterolactone concentration is affected by previous use of oral antimicrobials. Enterolactone was measured by time-resolved fluoroimmunoassay in 2,753 Finnish men and women aged 25--64 years who participated in a cross-sectional national survey in 1997. Background information was collected by self-administered questionnaire, and data on antimicrobial treatment were gathered from the nationwide prescription database of the Social Insurance Institution. Serum enterolactone concentration was significantly lower in those who had used oral antimicrobials up to 12--16 months before serum sampling than in nonusers (16.4 vs. 19.3 nmol/liter). The concentration was associated with the number of treatments and the time from the last treatment. Modest differences were present between various antimicrobials. The authors' findings support the crucial role of gut microflora in the metabolism of lignans. Furthermore, recent use of antimicrobials should be considered when the association between serum enterolactone concentration and risk of chronic diseases is studied. PMID- 11867361 TI - Rapacuronium redux. PMID- 11867360 TI - Sample size requirements for association studies of gene-gene interaction. AB - In the study of complex diseases, it may be important to test hypotheses related to gene-gene (G x G) interaction. The success of such studies depends critically on obtaining adequate sample sizes. In this paper, the author investigates sample size requirements for studies of G x G interaction, focusing on four study designs: the matched-case-control design, the case-sibling design, the case parent design, and the case-only design. All four designs provide an estimate of interaction on a multiplicative scale, which is used as a unifying theme in the comparison of sample size requirements. Across a variety of genetic models, the case-only and case-parent designs require fewer sampling units (cases and case parent trios, respectively) than the case-control (pairs) or case-sibling (pairs) design. For example, the author describes an asthma study of two common recessive genes for which 270 matched case-control pairs would be required to detect a G x G interaction of moderate magnitude with 80% power. By comparison, the same study would require 319 case-sibling pairs but only 146 trios in the case-parent design or 116 cases in the case-only design. A software program that computes sample size for studies of G x G interaction and for studies of gene-environment (G x E) interaction is freely available (http://hydra.usc.edu/gxe). PMID- 11867362 TI - Cerebral hyperemia, systemic hypertension, and perioperative intracranial morbidity: is there a smoking gun? PMID- 11867363 TI - Rapacuronium and the risk of bronchospasm in pediatric patients. AB - We conducted this study to determine the risk factors for the development of bronchospasm after the administration of rapacuronium and to determine if children with bronchospasm on induction of anesthesia were more likely to have received rapacuronium compared with other muscle relaxants. In a retrospective cohort study, all anesthetic records in which rapacuronium was administered were reviewed to determine which patients developed bronchospasm during induction of anesthesia. Two-hundred-eighty-seven patients were identified, of whom 12 (4.2%; 95% confidence interval [CI], 2.2%--7.2%) developed bronchospasm during induction of anesthesia. Significant risk factors for the development of bronchospasm with administration of rapacuronium included rapid sequence induction (relative risk [RR], 17.9; 95% CI, 2.9--infinity) and prior history of reactive airways disease (RR, 4.6; 95% CI, 1.5--14.3). In a case-control study, all cases of bronchospasm during induction of anesthesia in the 5-mo time period that rapacuronium was available for clinical use were identified. Aside from the 12 cases of bronchospasm with rapacuronium, 11 additional cases of bronchospasm were associated with the use of other muscle relaxants. Four controls were randomly selected for each of the 23 cases of bronchospasm. Children with bronchospasm during induction of anesthesia were several times more likely (odds ratio, 10.1; 95% CI, 3.5--28.8) for having received rapacuronium compared with other muscle relaxants. IMPLICATIONS: In a retrospective cohort study, significant risk factors for the development of bronchospasm with the administration of rapacuronium on induction of anesthesia included rapid sequence induction and prior history of reactive airways disease. In a case-control study, children with bronchospasm during induction of anesthesia were several times more likely to have received rapacuronium compared with other muscle relaxants. PMID- 11867364 TI - The effect of chin lift, jaw thrust, and continuous positive airway pressure on the size of the glottic opening and on stridor score in anesthetized, spontaneously breathing children. AB - Chin lift and jaw thrust are two common maneuvers used to improve the patency of the upper airway during general anesthesia. We investigated the effect of these maneuvers combined with continuous positive airway pressure (CPAP) on the size of glottic opening and on stridor score. Forty children, aged 2--9 yr, premedicated with midazolam and spontaneously breathing end-tidal 1% halothane and equal parts of nitrous oxide and oxygen, were studied. A flexible fiberoptic bronchoscope was placed via mask and one nostril to the level of the junction of the soft palate and oropharynx. Video recordings and simultaneous stridor scores were obtained during six conditions: 1) chin unsupported, 2) manual chin lift, 3) chin lift and CPAP 10 cm H(2)O, 4) repeat chin unsupported, 5) manual jaw thrust, and 6) jaw thrust and CPAP 10 cm H(2)O. Videos were analyzed to determine the percentage of glottic opening (POGO) score. POGO score increased (P < 0.05) in Conditions 2, 3, 5, and 6. With increasing POGO score there was a decrease in stridor score (P < 0.05). IMPLICATIONS: Chin lift and jaw thrust maneuvers combined with continuous positive airway pressure improve the view of the glottic opening as viewed by flexible nasal laryngoscopy and decrease stridor in anesthetized, spontaneously breathing children. PMID- 11867365 TI - The effects of low-pressure carbon dioxide pneumoperitoneum on cerebral oxygenation and cerebral blood volume in children. AB - We examined the effects of low-pressure carbon dioxide pneumoperitoneum on regional cerebral oxygen saturation (ScO(2)) and cerebral blood volume (CBV) in children. Fifteen children, ASA I--III, scheduled for laparoscopic fundoplication, were investigated in the head-up position (10) and ventilated to a baseline end-tidal CO(2) (PETCO(2)) between 25 and 33 mm Hg. Ventilatory settings remained unchanged during the operation. ScO(2) and CBV were assessed with near-infrared spectroscopy and recorded together with end-tidal and arterial carbon dioxide (PaCO(2)) at 5 time points: before insufflation, 30, 60, and 90 min after the start of CO(2) insufflation, and 10 min after desufflation. The intraabdominal pressure was kept between 5 and 8 mm Hg. During insufflation, PETCO(2) increased from 30.0 plus minus 2.8 to 38.3 plus minus 5.1 mm Hg (P < 0.001) and PaCO(2) increased from 32.0 plus minus 4.7 to 40.4 plus minus 5.9 mm Hg (P < 0.001). ScO(2) increased by 15.7% plus minus 8.8% (from 61 plus minus 9 to 70 plus minus 9 arbitrary units ) (P < 0.001). CBV increased by 4.6% plus minus 8.8% (from 123 plus minus 66 to 128 plus minus 66 arbitrary units [P = 0.048]). After desufflation, PETCO(2) and PaCO(2) decreased, but did not return to preinsufflation values. ScO(2) and CBV also decreased after desufflation. In conclusion, hyperventilation and the head-up position before CO(2) insufflation are not sufficient to prevent the CO(2)-mediated cerebral hemodynamic effects of low-pressure pneumoperitoneum (5--8 mm Hg) in children. IMPLICATIONS: Peritoneal CO(2) absorption during laparoscopic surgery causes hypercapnia and CO(2) mediated cerebral hemodynamic effects. Hyperventilation and the head-up position before CO(2) insufflation is not sufficient to counteract these effects of low pressure pneumoperitoneum (5--8 mm Hg) in children. PMID- 11867366 TI - The correlation of the bispectral index monitor with clinical sedation scores during mechanical ventilation in the pediatric intensive care unit. AB - In patients who are mechanically ventilated in the pediatric intensive care unit (PICU), sedative and/or analgesic medications are routinely provided and titrated to effect based on clinical assessment of the patient. The bispectral index (BIS) monitor uses a modified electroencephalogram to quantify the effects of central nervous system-acting drugs on the level of consciousness. To evaluate the usefulness of the BIS monitor to predict clinical sedation levels in the PICU, we compared BIS values with simultaneously obtained clinical sedation scores in 24 mechanically ventilated pediatric patients aged 5.7 plus minus 6.1 yr. For each sedation scale used, the BIS value correlated with increasing depth of sedation (P < 0.0001) and was independent of the drug(s) used for sedation. To differentiate adequate from inadequate sedation, a BIS value <70 had a sensitivity of 0.87--0.89 and a positive predictive value of 0.68--0.84. To differentiate adequate from excessive sedation, a BIS value <50 had a sensitivity of 0.67--0.75 and a positive predictive value of 0.07--0.52. We conclude that the BIS monitor may be a useful adjunct for the assessment of sedation in PICU patients. IMPLICATIONS: We demonstrate the usefulness of the bispectral index monitor for assessing sedation in pediatric intensive care unit patients. The bispectral index monitor correlated with clinically assessed sedation levels and was useful for differentiating adequate from inadequate sedation, which would be of value when the clinical examination is unavailable. PMID- 11867367 TI - Tension pneumothorax during flexible fiberoptic bronchoscopy in a newborn. AB - IMPLICATIONS: A newborn undergoing flexible fiberoptic bronchoscopy before repair of a tracheoesophageal fistula developed a life-threatening tension pneumothorax. The mechanisms of this complication are discussed and recommendations are made for preventing this complication. PMID- 11867368 TI - The pharmacokinetics and cardiovascular effects of a single intravenous dose of protamine in normal volunteers. AB - Despite its long use in clinical medicine, protamine concentrations and pharmacokinetics in humans have not been reported. The occasional reoccurrence of anticoagulation after protamine reversal of heparin led us to hypothesize that protamine plasma concentrations decrease rapidly. We developed a method for the measurement of protamine in plasma. Eighteen fit volunteers gave their consent to receive 0.5 mg/kg protamine sulfate administered IV by an infusion pump over 10 min. Heart rate, mean arterial blood pressure, and cardiac output, all measured noninvasively, were recorded and blood samples obtained during and after protamine infusion. Blood plasma was subjected to solid-phase extraction and high performance liquid chromatography. The administration of protamine was associated with no significant changes in heart rate, mean arterial blood pressure, or cardiac output. Plasma protamine concentrations decreased rapidly, becoming nondetectable within approximately 20 min. Protamine elimination differed significantly between men and women: men had significantly larger areas under the concentration versus time curve. Model-independent pharmacokinetic analysis revealed median (range) values as follows: volume of distribution at steady state, 12.3 (6.9--63.1) L; clearance, 2.2 (1.1--12.1) L/min; and t1/2, 7.4 (5.9- 9.3) min. Concentration versus time plots revealed an atypical pattern inconsistent with usual exponential models. The Schwartz-Bayesian criterion identified a one-compartment Michaelis-Menten model and a two-compartment exponential model with irreversible binding as performing better than conventional one- or two-compartmental exponential models; however, performance errors were large with both Michaelis-Menten and exponential models. All models described rapid decreases in protamine blood concentrations. IMPLICATIONS: We developed a method for measurement of protamine in human blood. In volunteers, protamine concentrations decreased rapidly after administration. The rapid disappearance of protamine from the circulation, as defined by a median half-life of 7.4 min, could contribute to cases of "heparin rebound" after initial adequate reversal of heparin. PMID- 11867369 TI - Tramadol infusion for postthoracotomy pain relief: a placebo-controlled comparison with epidural morphine. AB - We compared continuous IV tramadol as an alternative to neuraxial or systemic opioids for the management of postthoracotomy pain in a prospective, randomized, double-blinded, controlled study. General anesthesia was supplemented by thoracic epidural analgesia with 0.25% bupivacaine. At rib approximation, patients received one of the following: IV tramadol (150-mg bolus followed by infusion, total 450 mg/24 h, n = 29), epidural morphine (2 mg, then 0.2 mg/h, n = 30), or patient-controlled analgesia (PCA) morphine only (n = 30). All patients received PCA morphine and rescue morphine as necessary postoperatively. For the first 24 h, pain and sedation scores and respiratory, cardiovascular, and side effect measures were monitored. There was no significant difference in pain scores and PCA morphine use between tramadol and epidural morphine. Pain scores at rest and on coughing were lower in the Tramadol and Epidural Morphine groups than in the PCA Morphine group at various time points over the first 12 h. The Tramadol and Epidural Morphine groups used significantly less hourly PCA morphine than the PCA Morphine group at specific time points in the first 10 h. Vital capacities in the Tramadol group were significantly closer to baseline values at the 20-h point than in the PCA Morphine group. We conclude that an intraoperative bolus of tramadol followed by an infusion was as effective as epidural morphine and avoided the necessity of placing a thoracic epidural catheter. IMPLICATIONS: A prospective, randomized, double-blinded, placebo-controlled study of postthoracotomy pain relief showed that IV tramadol in the form of a bolus followed by continuous infusion was as effective as epidural morphine. The use of tramadol avoids the necessity of placing a thoracic epidural catheter. PMID- 11867370 TI - The effect of universal leukodepletion of packed red blood cells on postoperative infections in high-risk patients undergoing abdominal aortic surgery. AB - We evaluated, by using a before-and-after study, the influence of leukoreduction by filtration on postoperative infections and adverse outcomes in patients undergoing elective major aortic surgery. From January 1995 to October 2000, all patients who underwent elective abdominal aortic surgery were included in the analysis. Before the introduction of systematic leukodepletion of packed red blood cells (RBCs), on April 1, 1998, 192 patients received standard or buffy coat-depleted packed RBCs. Then, 195 patients were transfused with exclusively filtered leukodepleted packed RBCs. No major significant difference was observed between the groups of patients with regard to preoperative cardiac and pulmonary status, anesthetic and surgical techniques, or transfusion policy. No significant difference in mortality was observed between the two groups. The incidence of postoperative infections was 31% (95% confidence interval, 25%--38%) in the Control group versus 27% (95% confidence interval, 21%--33%) in the Leukodepleted group; severe infectious complications and pneumonia were not significantly different between the two groups of patients. Cardiovascular and respiratory outcomes were not significantly different between the groups. Data from this study suggest that the effect of using leukodepleted RBC on postoperative infections is not of obvious importance. IMPLICATIONS: We evaluated the influence of leukocyte reduction by filtration of packed red blood cells (RBC) on postoperative infections and adverse outcomes in patients undergoing elective major aortic surgery by comparing two epochs with and without filtration. Data from this study suggest that the effect of using filtered RBC on postoperative infections is not of obvious importance. PMID- 11867371 TI - Hydroxyethyl starch in balanced electrolyte solution (Hextend)--pharmacokinetic and pharmacodynamic profiles in healthy volunteers. AB - Hextend is a new plasma volume expander containing 6% hydroxyethyl starch (HES) in a physiologically balanced medium of electrolytes, glucose, and lactate (weight average, molecular weight 670 kDa, molar substitution 0.75). This open label study was designed to investigate the pharmacokinetic and pharmacodynamic profiles of Hextend in 21 healthy volunteers. We infused Hextend 10 ml/kg IV over 20 min and determined serum concentrations of HES at selected intervals over a 7 day period. Serum concentration-time curves indicated mixed pharmacokinetic behavior reflecting a two-compartment model in most subjects. The median serum half-life over 7 days was 38.2 h. The balanced formulation of the suspension medium did not seem to affect distribution, metabolism, or excretion of Hextend when compared with similar HES. Pharmacodynamic analysis demonstrated decreases in some plasma components compatible with the infusion of that volume of fluid and the duration of plasma volume expansion. Other plasma components remained unchanged, reflecting the benefit of a balanced electrolyte solution. Hemodilution was observed for 24--48 h after short-term infusion of Hextend. Some hemostatic indices showed moderate changes, and serum amylase demonstrated a temporary increase. Our study suggested that Hextend has pharmacokinetic and pharmacodynamic profiles that are similar to those of other HES. IMPLICATIONS: Hextend is a new plasma volume expander containing 6% hydroxyethyl starch in a physiologically balanced medium. This open-label volunteer study demonstrated that it has pharmacokinetic and pharmacodynamic profiles similar to those of established HES. PMID- 11867372 TI - The direct myocardial effects of xenon in the dog heart in vivo. AB - Xenon has minimal hemodynamic side effects, but no data are available on its direct myocardial effects in vivo. We examined myocardial function during the global and regional administration of xenon in the dog heart. Anesthetized (midazolam/piritramide) dogs (n = 8) were instrumented for measurement of left ventricular pressure, cardiac output, and blood flow in the left anterior descending coronary artery (LAD) and circumflex coronary artery. Regional myocardial function was assessed by sonomicrometry in the antero-apical and the postero-basal wall. Hemodynamics were recorded during baseline conditions and during inhalation of 50% or 70% xenon, respectively. Subsequently, a bypass containing a membrane oxygenator was installed from the carotid artery to the LAD, allowing xenon administration only to the LAD-dependent myocardium. No changes in myocardial function were observed during inhalation of xenon. The regional administration of 50% xenon had no significant effect on regional myocardial function (systolic wall thickening and mean velocity of systolic wall thickening). Seventy percent xenon reduced systolic wall thickening by 7.2% +/- 4.0% and mean velocity of systolic wall thickening by 8.2% +/- 4.0% in the LAD perfused area (P < 0.05). There were no changes of global hemodynamics, coronary blood flow, and regional myocardial function in the circumflex coronary artery dependent myocardium. Xenon produces a small but consistent direct negative inotropic effect in vivo. IMPLICATIONS: Regional administration of xenon direct to the left anterior descending-perfused myocardium resulted in a small but consistent negative inotropic effect of the noble gas in the dog heart in vivo. PMID- 11867373 TI - Ketamine suppresses norepinephrine-induced inositol 1,4,5-trisphosphate formation via pathways involving protein kinase C. AB - Inositol 1,4,5-trisphosphate (IP(3)) is not only involved in the physiologic regulation of excitation-contraction coupling, but could also play a role in cardiac pathophysiology. We investigated the mechanism of ketamine modulation of norepinephrine (NE)-induced IP(3) formation in neonatal rat cardiomyocytes. Ketamine 1 and 10 microM significantly decreased the IP(3) response to 1 microM NE by 27% and 43%, respectively. One micromolar TMB-8 (an intracellular calcium antagonist) produced 42% more decreases in IP(3) production than produced by ketamine alone. One hundred micromolar anthranilic acid (a phospholipase A(2) inhibitor) significantly decreased NE (1 microM)-induced IP(3) formation, and the inhibition was further enhanced by ketamine. Ten micromolar U 73122 (a phospholipase C inhibitor) did not significantly affect NE-induced IP(3) in the presence or absence of ketamine. One micromolar ketamine significantly inhibited staurosporine (a nonselective protein kinase C antagonist)-, bisindolylmaleimide (a selective protein kinase C antagonist)-, and wortmannin (a phosphatidylinositide 3-kinase antagonist)-stimulated IP(3) formation. In conclusion, ketamine suppresses NE-induced IP(3) production, and the inhibition is caused through pathways including protein kinase C and a decrease in intracellular Ca(2+) concentrations. IMPLICATIONS: Ketamine inhibits norepinephrine-induced inositol 1,4,5-triphosphate formation in a dose-dependent manner via pathways that involve protein kinase C and a decrease in intracellular Ca(2+) concentrations. PMID- 11867374 TI - Dynamic left ventricular outflow tract obstruction complicating bilateral lung transplantation. AB - Dynamic left ventricular outflow tract obstruction (DLVOTO) has been observed in a variety of clinical circumstances but not previously reported in the setting of orthotopic lung transplantation. Diagnosis and effective management of this adverse event were facilitated by transesophageal echocardiography (TEE). IMPLICATIONS: Dynamic left ventricular outflow tract obstruction is not an uncommon phenomenon, but it is often unrecognized. Its rapid recognition and effective treatment was only possible with transesophageal echocardiography (TEE). This contributes to the support for TEE being routinely available for assessment of hemodynamic instability. PMID- 11867375 TI - Fast-track eligibility of geriatric patients undergoing short urologic surgery procedures. AB - Our primary objective was to assess the feasibility of geriatric patients (>65 yr) bypassing the postanesthesia care unit (PACU) after ambulatory surgery. A secondary objective was to compare recovery profiles when using three different maintenance anesthetics. Ninety ASA physical status I--III consenting outpatients (>65 yr) undergoing short urologic procedures were randomly assigned to one of three anesthetic treatment groups. After a standardized induction with fentanyl and propofol, anesthesia was maintained with propofol (75-150 microg center dot kg(-1) center dot min(-1) IV), isoflurane (0.7%-1.2% end tidal), or desflurane (3%-6% end tidal), in combination with nitrous oxide 70% in oxygen. In all three groups, the primary anesthetic was titrated to maintain an electroencephalographic-bispectral index value of 60-65. Recovery times, postanesthesia recovery scores, and therapeutic interventions in the PACU were recorded. Although emergence times were similar in the three groups, the time to achieve a fast-track discharge score of 14 was significantly shorter in patients receiving desflurane compared with propofol and isoflurane (22 +/- 23 vs 33 +/- 25 and 44 +/- 36 min, respectively). On arrival in the PACU, a significantly larger percentage of patients receiving desflurane were judged to be fast-track eligible compared with those receiving either isoflurane and propofol (73% vs 43% and 44%, respectively). The number of therapeutic interventions in the PACU was also significantly larger in the Isoflurane group when compared with the Propofol and Desflurane groups (21 vs 11 and 7, respectively). In conclusion, use of desflurane for maintenance of anesthesia should facilitate PACU bypass ("fast tracking") of geriatric patients undergoing short urologic procedures. IMPLICATIONS: Geriatric outpatients undergoing brief urologic procedures more rapidly achieve fast-tracking discharge criteria after desflurane (versus isoflurane and propofol) anesthesia. Use of isoflurane was also associated with an increased need for nursing interventions in the early recovery period compared with desflurane and propofol. PMID- 11867376 TI - Selective spinal anesthesia versus desflurane anesthesia in short duration outpatient gynecological laparoscopy: a pharmacoeconomic comparison. AB - We compared the cost and effectiveness of selective spinal anesthesia (SSA) with a desflurane-based general anesthetic (DES) for outpatient gynecological laparoscopy. A prospective analysis was undertaken of 10 patients randomized to receive SSA and compared with 10 patients randomized to receive DES. The groups were well matched in their demographic characteristics. The mean cost (in 2000 Canadian dollar values) of anesthesia supplies, drugs, and nursing for the SSA group of $62.31 was less than that for the DES group of $92.31 (P < 0.01). Recovery costs of both groups were similar. Time to administer anesthesia and time spent in the postanesthetic care unit were also similar. Postoperative analgesia was required by 50% of the DES group but in no patient receiving SSA (P < 0.01). SSA is a cost-effective alternative to DES in these patients. IMPLICATIONS: Small-dose spinal anesthesia is an effective alternative to a desflurane general anesthetic in terms of cost and recovery profiles in ambulatory gynecological laparoscopy. PMID- 11867377 TI - Physostigmine does not antagonize sevoflurane anesthesia assessed by bispectral index or enhances recovery. AB - In this double-blinded study, we investigated the effect of physostigmine on sevoflurane anesthesia and recovery. Forty female patients scheduled for breast biopsy were randomly assigned to receive either physostigmine 2 mg IV or an equal volume of normal saline after skin closure. Anesthesia was induced and maintained with sevoflurane in oxygen. After skin closure, a steady state of 0.6% inspired and end-tidal sevoflurane concentration was obtained, heart rate, blood pressure, and Bispectral index (BIS) baseline values were recorded, and physostigmine or normal saline was administered. Hemodynamics and BIS values were also recorded 5, 8, and 10 min after treatments. Anesthesia was discontinued, and orientation, sedation, sitting ability, and "picking up matches" scores were recorded immediately after extubation and 15 and 30 min later. No differences were found between the two groups in BIS (69, 70, 70, and 71 in the Physostigmine group versus 70, 74, 75, and 76 in the Control group) or blood pressure. Only heart rate was increased 8 min after physostigmine (P < 0.05 versus the control). Scores assessing early recovery were similar in the two groups at all time points. We conclude that physostigmine does not change BIS or enhance recovery after sevoflurane anesthesia. IMPLICATIONS: This double-blinded, randomized study investigated the impact of physostigmine of BIS values during 0.6% sevoflurane anesthesia as well as in the postoperative recovery, when sevoflurane is administered as a sole anesthetic. Physostigmine has no effect on BIS values or on the tests assessing recovery. PMID- 11867378 TI - Pretreatment before succinylcholine for outpatient anesthesia? AB - IMPLICATIONS: This study demonstrated that pretreatment of succinylcholine with rocuronium failed to decrease the incidence or the severity of postoperative myalgia. However, in most patients, pretreatment was associated with muscle weakness before loss of consciousness. Thus, there is no convincing evidence supporting routine pretreatment with succinylcholine. PMID- 11867379 TI - The role of non-opioid analgesic techniques in the management of pain after ambulatory surgery. PMID- 11867380 TI - A comparison of the INVOS 4100 and the NIRO 300 near-infrared spectrophotometers. AB - We determined whether two different devices for measuring near-infrared spectroscopy (NIRS)---the INVOS 4100 and the NIRO 300---produce similar cerebral oxygenation data during the CO(2) challenge test. Nineteen patients anesthetized with sevoflurane, 67% nitrous oxide in oxygen, and fentanyl were studied. A series of measurements of regional cerebral oxygen saturation (rSO(2)), measured by the INVOS 4100, and tissue oxygen index (TOI), measured by the NIRO 300, were performed in the following conditions: 1) normocapnia (PaCO(2), 35--45 mm Hg); 2) hypocapnia (PaCO(2), 25--35 mm Hg); 3) normocapnia; and 4) hypercapnia (PaCO(2), 45--55 mm Hg). Hemodynamic variables, including arterial blood gases and cerebral blood flow velocity, were measured at the same time with transcranial Doppler. The values and percentage changes of rSO(2) and TOI were compared by using regression analysis and Bland and Altman analysis. The rSO(2) showed a significant positive correlation with TOI (r = 0.58, P < 0.01). The percentage change of rSO(2) also showed a significant positive correlation with the percentage change of TOI during the CO(2) challenge (r = 0.85, P < 0.01). Bland and Altman analysis revealed a bias of -0.5% with 2 SD of 15.6% when comparing the rSO(2) value with the TOI value, and it showed a bias of -3.4% with 2 SD of 15.2% when comparing the percentage change of rSO(2) with the percentage change of TOI, indicating unacceptable disagreement of these data. These results indicate that cerebral oxygen saturation and its relative change during the CO(2) challenge may vary depending on the type of NIRS used. Because the measurement technique and algorithm were different in each device, we should carefully consider the clinical application of the values produced by NIRS. IMPLICATIONS: Near-infrared spectroscopy (NIRS) has been proposed as a noninvasive clinical method for assessing cerebral oxygenation. The acceptable reliability and validity of NIRS values have not been established despite their widespread use. The INVOS 4100 and the NIRO 300 can display cerebral oxygen saturation as regional cerebral oxygen saturation and tissue oxygenation index, but they produce differing results. PMID- 11867381 TI - The neuromuscular transmission module versus the relaxometer mechanomyograph for neuromuscular block monitoring. AB - The neuromuscular transmission module (M-NMT) is an integrated piezoelectric motion sensor module incorporated in the AS/3(TM) anesthesia monitor. We compared the neuromuscular block of 0.6 mg/kg rocuronium (twice the 95% effective dose) monitored by the M-NMT with that monitored by the Relaxometer mechanomyograph (MMG). The two monitors were alternately allocated to the left or right hands of 20 patients. T(1)%, the first twitch of the train-of-four (TOF), and the TOF ratio (T(4)/T(1)) were used for evaluating the neuromuscular block. There was no significant difference in the mean (min) plus minus SD onset time or time to 0.8 TOF ratio recovery measured by the M-NMT (1.5 plus/minus 0.3, 49.4 plus/minus 8.1) compared with MMG (1.8 plus/minus 0.6, 50.9 plus/minus 9.9), respectively. However, the time (min) to 25% T(1) recovery was significantly longer when monitored by the M-NMT (25.6 plus/minus 8) than by the MMG (20.2 plus/minus 6.3). During recovery from neuromuscular block, the difference between the TOF ratios measured by the two monitors showed a bias of -0.031, and the limits of agreement (bias plus/minus 1.96 SD) were -0.281 and +0.22. The M-NMT monitor could determine the time to tracheal intubation as well as full recovery from neuromuscular block, but it lagged behind the MMG in determining the time to rocuronium repeat dose administration. IMPLICATIONS: Compared with the Relaxometer mechanomyograph, the neuromuscular transmission module could equally indicate time to tracheal intubation and full recovery from 0.6 mg/kg rocuronium neuromuscular block. Its small quick-fit sensor has the advantage, in an often crowded and busy operating room, of being incorporated in the AS/3(TM) anesthesia workstation. PMID- 11867382 TI - The potency of new muscle relaxants on recombinant muscle-type acetylcholine receptors. AB - We studied the inhibition of fetal (gamma-nAChR) and adult (epsilon-nAChR) muscle type nicotinic acetylcholine receptors by the two new nondepolarizing muscle relaxants (NDMRs) rocuronium and rapacuronium, the metabolite 3-desacetyl rapacuronium (Org 9488), and five other, longer-used NDMRs (pancuronium, vecuronium, mivacurium, d-tubocurarine, and gallamine). Receptors were expressed in Xenopus laevis oocytes by cytoplasmic injection of subunit complementary RNAs. Functional channels were activated with 10 microM acetylcholine, alone or in combination with various concentrations of the NDMRs. Currents were recorded with a whole-cell two-electrode voltage clamp technique. All NDMRs reversibly inhibited acetylcholine-activated currents in a dose-dependent fashion. Potencies of rapacuronium and Org 9488 were not statistically different at either gamma nAChR (half-maximal response = 58.2 and 36.5 nM, respectively) or epsilon-nAChR (half-maximal response = 80.3 and 97.7 nM, respectively). The rank order of potencies at the epsilon-nAChR (pancuronium > vecuronium similar mivacurium > rocuronium similar d-tubocurarine > rapacuronium similar Org 9488 > gallamine) correlated highly with the clinical doses needed to produce 50% twitch depression at the adductor pollicis muscle in adults. Neuromuscular blockade by rapacuronium may be enhanced by its metabolite Org 9488. Different drug-receptor affinities of the tested NDMRs contribute to the differences in clinical dose requirements of these drugs needed to achieve appropriate muscle relaxation. IMPLICATIONS: Potencies of nondepolarizing muscle relaxants, studied at muscle nicotinic acetylcholine receptors expressed in a recombinant expression system, correlate highly with the clinical doses needed in adults to produce 50% twitch depression at the adductor pollicis muscle. PMID- 11867384 TI - Oral clonidine premedication reduces the awakening concentration of propofol. AB - To investigate the effects of oral clonidine premedication on emergence from propofol/fentanyl anesthesia, we studied 72 healthy male patients who were undergoing elective orthopedic surgery: the Control group, the 2.5 microg/kg Clonidine group, and the 5.0 microg/kg Clonidine group (n = 24 each). Nothing was administered to the Control group. Clonidine (2.5 or 5.0 microg/kg) was orally administered 90 min before the induction of anesthesia in the Clonidine groups. Patients were anesthetized with computer-assisted continuous infusion of propofol and fentanyl, with the three groups receiving the same concentrations of propofol (3 microg/mL) and fentanyl (1 ng/mL) starting 20 to 30 min before the end of surgery. Propofol infusion was then abruptly discontinued at the end of surgery in all patients. After propofol was discontinued, the response to verbal commands was evaluated every 30 s, and arterial blood samples for propofol and clonidine concentrations were taken when the patients opened their eyes. The time required to respond to a verbal command was 14.9 plus/minus 8.3 min for the 5.0 microg/kg Clonidine group, and this was significantly longer than the Control (8.2 plus/minus 5.0 min) and the 2.5 microg/kg Clonidine (9.0 plus/minus 3.7 min) groups (P < 0.01). Serum propofol concentration at awakening in the 5.0 microg/kg Clonidine group was 1.0 plus/minus 0.4 microg/mL, which was significantly smaller than the Control (1.6 plus/minus 0.4 microg/mL) and the 2.5 microg/kg Clonidine (1.4 plus/minus 0.3 microg/mL) groups (P < 0.01). The blood clonidine concentration was associated with a decrease in the awakening propofol concentration. In conclusion, 5 microg/kg oral clonidine premedication decreases the awakening propofol concentration and delays arousal from propofol/fentanyl anesthesia. IMPLICATIONS: Preanesthetic medication with 5 microg/kg oral clonidine, but not 2.5 microg/kg clonidine, is associated with prolonged recovery from propofol/fentanyl anesthesia. PMID- 11867383 TI - Muscle relaxation does not alter hypnotic level during propofol anesthesia. AB - Electromyographic (EMG) activity can contaminate electroencephalographic signals. Paralysis may therefore reduce the Bispectral Index (BIS) by alleviating artifact from muscles lying near the electrodes. Paralysis may also reduce signals from muscle stretch receptors that normally contribute to arousal. We therefore tested the hypothesis that nondepolarizing neuromuscular block reduces BIS. Ten volunteers were anesthetized with propofol at a target effect site concentration of 3.8 plus/minus 0.4 microg/mL. A mivacurium infusion was adjusted to vary the first twitch (T1) in a train-of-four to 80%, 30%, 20%, 15%, 10%, 5%, or 2% of the prerelaxant intensity. At each randomly assigned T1, we measured BIS and frontal temporal EMG intensity. BIS averaged 95 plus/minus 4 before induction of anesthesia, and decreased significantly to 40 plus/minus 5 after propofol administration. However, there were no significant differences at the designated block levels. Frontal-temporal EMG intensity averaged 47 plus/minus 3 dB before induction of anesthesia, and decreased significantly to 27 plus/minus 1 dB after propofol administration. However, there were no significant differences at the designated block levels. These data suggest that the BIS level and EMG tone are unaltered by mivacurium administration during propofol anesthesia. IMPLICATIONS: Neuromuscular block level did not alter Bispectral Index (BIS) during propofol anesthesia, either by reducing electromyographic artifact or by decreasing afferent neuronal input. The BIS will thus comparably estimate sedation in deeply unconscious patients who are paralyzed, partially paralyzed, or unparalyzed. PMID- 11867385 TI - Preoperative clonidine blunts hyperadrenergic and hyperdynamic responses to prolonged tourniquet pressure during general anesthesia. AB - Although the mechanism of tourniquet-induced hypertension is still unclear, plasma norepinephrine concentrations continuously increase in parallel to arterial blood pressure during tourniquet inflation. Clonidine attenuates hyperadrenergic and hyperdynamic responses. We investigated the effects of clonidine on prolonged tourniquet inflation. Twenty-nine patients scheduled for elective orthopedic surgery were randomly assigned to receive IV clonidine (3 microg/kg; n = 14) or placebo (n = 15) before tourniquet inflation of the lower limbs under general anesthesia in a double-blinded manner. Arterial blood pressure, heart rate, epinephrine, and norepinephrine plasma concentrations were measured before tourniquet inflation, 60 min after tourniquet inflation, just before tourniquet deflation, and 20 min after tourniquet deflation. Mean arterial blood pressure and norepinephrine plasma-concentrations were significantly lower in the Clonidine group compared with Control after 60 min tourniquet inflation (P = 0.016; P = 0.006). Immediately before deflation of the tourniquet, the difference for mean arterial pressure between groups was even more pronounced (P = 0.005). Twenty minutes after deflation mean arterial blood pressure in the Control group was still increased and significantly higher compared with the Clonidine group (P = 0.002). In conclusion, preoperative IV clonidine blunts hyperadrenergic and hyperdynamic responses resulting from prolonged tourniquet inflation under general anesthesia in ASA class I--II patients. IMPLICATIONS: Tourniquet inflation is associated with a continuous increase in arterial blood pressure and sympathetic outflow. This study shows that IV clonidine effectively blunts increases of both arterial blood pressure and plasma norepinephrine concentrations. PMID- 11867386 TI - Tramadol has no effect on cortical renal blood flow--despite increased serum catecholamine levels--in anesthetized rats: implications for analgesia in renal insufficiency. AB - Tramadol is an analgesic that inhibits norepinephrine (NE) reuptake. Although NE released from renal sympathetic nerves causes renal hypoperfusion, the effects of tramadol on renal hemodynamics have not been well characterized. We investigated the effects of tramadol on renal blood flow (RBF), mean arterial blood pressure (MAP), and heart rate (HR) by using a laser Doppler flowmeter, both in normal anesthetized rats and in rats with experimentally-induced nephritis secondary to anti-Thy 1.1 antibody administration. We also studied the effects of tramadol on serum NE levels. Tramadol increased MAP and decreased HR without changing RBF in normal rats at clinical doses. Serum NE levels increased up to 176% of control after a 2 mg/kg bolus injection of tramadol. Continuously infused, increasing doses of tramadol (0.5-4 mg.kg(-1).h(-1)) did not affect MAP, HR, or RBF. Tramadol also increased MAP and decreased HR without changing RBF in rats with experimentally induced renal insufficiency. These findings suggest that a bolus injection of tramadol does not alter RBF, although it causes a decrease in HR and an increase in MAP and serum NE in both normal rats and in rats with renal insufficiency. These results suggest that tramadol may have little effect on RBF during the postoperative period. IMPLICATIONS: A bolus and continuous injection of tramadol does not alter renal blood flow (RBF) in normal rats. A bolus injection of tramadol has little effect on RBF in rats with experimentally induced renal insufficiency. These results suggest that tramadol would be a safe analgesic for maintaining RBF during the postoperative period. PMID- 11867387 TI - The effects of urapidil on thermoregulatory thresholds in volunteers. AB - In a previous study we have shown that the antihypertensive drug, urapidil, stops postanesthetic shivering. One possible mechanism in the inhibition of postanesthetic shivering by urapidil may be alterations in thermoregulatory thresholds. We therefore studied the effects of urapidil on vasoconstriction and shivering thresholds during cold-induced shivering in volunteers. Seven healthy male volunteers were cooled by an infusion of saline at 4 degrees C on two study days separated by 48 h. Thermoregulatory vasoconstriction was estimated using forearm minus fingertip skin-temperature gradients, and values exceeding 0 degrees C were considered to represent significant vasoconstriction. The rectal core temperatures at the beginning of shivering and at vasoconstriction were considered the thermoregulatory thresholds. Before cooling, either 25 mg of urapidil or placebo was administered randomly and blindly to each volunteer. When shivering occurred continuously for 10 min, another 25 mg of urapidil was administered IV to completely stop shivering. Urapidil led to a decrease in core temperature at vasoconstriction and shivering threshold by 0.4 degrees C plus/minus 0.2 degrees C (P < 0.001) and 0.5 degrees C plus/minus 0.3 degrees C (P < 0.01), respectively. Oxygen consumption increased during shivering by 70% plus/minus 30% (P < 0.01) in comparison with baseline and decreased levels after shivering stopped, despite the continued low core temperature. Our investigation shows that urapidil stops postanesthetic shivering by decreasing important thermoregulatory thresholds. This means that shivering, not hypothermia, is treated, and hypothermia will need more attention in the postanesthesia care unit. IMPLICATIONS: In this study we show that the antihypertensive drug urapidil stops cold-induced shivering and decreases normal thermoregulatory responses, i.e., the thresholds for vasoconstriction and shivering, in awake volunteers. PMID- 11867388 TI - Short-term memory resists the depressant effect of the nonimmobilizer 1-2 dichlorohexafluorocyclobutane (2N) more than long-term memory. AB - The nonimmobilizer 1,2-dichlorohexafluorocyclobutane (2N, also termed F6) does not suppress movement to noxious stimuli but does suppress learning of fear potentiated startle. The mechanism whereby 2N suppresses this learning is unknown. Herein, we report the effect of 2N on suppression of two other forms of learning, fear conditioning to context and to tone. Because 2N does not cause sedation, we could study the effect of 2N on short-term memory (memory for fear conditioning measured during or immediately after training) as well as on long term memory (measured 24 h after training). The EC(50) for suppression of long term memory (the concentration decreasing memory by 50%) of fear conditioning to context was 2.00% plus/minus 0.01% (mean plus/minus SEM), and for fear conditioning to tone was 3.45% plus/minus 0.26%, (P < 0.05). The EC(50) for suppression of short-term memory of fear conditioning to context was 2.59% plus/minus 0.21% (P < 0.05, compared with long-term memory of context conditioning), whereas short-term memory of fear conditioning to tone was not suppressed by 3.5%, the largest concentration studied. Thus, short-term memory resists the depressant effect of 2N more than long-term memory, fear conditioning to tone is less vulnerable to the effect of 2N than fear conditioning to context, and 3.5% 2N does not preclude transmission of tone and shock signals to the site where tone-shock associations are formed. IMPLICATIONS: The nonimmobilizer 1,2 dichlorohexafluorocyclobutane has a greater depressant effect on long-term memory than short-term memory, suggesting that it impairs the processes responsible for the retention of memory more than for the formation of memory itself. PMID- 11867389 TI - Statistical power analysis to estimate how many months of data are required to identify operating room staffing solutions to reduce labor costs and increase productivity. AB - We performed a statistical power analysis to determine how many historical data are needed for optimal operating room (OR) management decision making. The work applies to hospitals that provide service for all of its surgeons' elective cases on whatever workday the surgeons and patients choose. The hospital and anesthesia group adjust OR staffing and patient scheduling to care for the patients while minimizing OR staffing costs and maximizing labor productivity. Two years of data were obtained from a seven-OR surgical suite. The data were repeatedly split into training and testing datasets. The optimal staffing solution was calculated for each training dataset to maximize the efficiency of OR time usage and was then applied to the corresponding testing dataset. Training datasets ranged in size from 30 to 270 consecutive workdays. With 30 workdays of data, the statistical method identified staffing solutions that had an average of 35% decreased costs and 27% increased productivity as compared to the existing staffing plan. There was no significant improvement in performance with more than 210 workdays (10 mo) of data. With 30 workdays of OR or anesthesia group data, the optimization method can significantly reduce staffing costs and increase productivity compared with existing staffing. When applied routinely for adjusting staffing (e.g., on a quarterly basis), 9 to 12 mo of data should be used. IMPLICATIONS: With 30 workdays of operating room or anesthesia group data, the optimization method can propose staffing solutions that significantly decrease costs and increase productivity compared with existing staffing solutions. We recommend that, when the statistical method is applied routinely for adjusting staffing (e.g., on a quarterly basis), 9 to 12 mo of data be used. PMID- 11867390 TI - The effect of outpatient preoperative evaluation of hospital inpatients on cancellation of surgery and length of hospital stay. AB - To evaluate the possible effects of outpatient preoperative evaluation (OPE) for new surgical patients who will be inpatients, we conducted an observational study at a university hospital in The Netherlands. Various outcomes before and after the introduction of an OPE clinic were compared. The study population comprised all 21,553 elective adult inpatients operated on between January 1, 1997 and December 31, 1999. Cardiac surgery, obstetric and pediatric patients, and patients operated on in same-day surgery were excluded. The main outcome measures were surgical cases canceled for medical reasons, rate of same-day admissions (who were expected to increase), and length of hospital stay. After introduction of OPE, the rate of cancellations for medical reasons decreased from 2.0% to 0.9% (adjusted odds ratio 0.7, 95% CI, 0.5--0.9). The rate of same-day admissions increased from 5.3% before to 7.7% after OPE introduction (adjusted odds ratio 1.2, 95% CI, 1.01--1.39), and the total hospital length of stay (in days) significantly decreased by a factor of 0.92 (0.90--0.94), which was partly the result of a reduction in preoperative admission time. We concluded that, although smaller than anticipated, the use of OPE for potential inpatients leads to a significant reduction of cancelled cases and of length of admission. Further increase of these benefits from OPE requires changes in institutional policy, such as forcing surgical departments to increase their number of same-day admissions. IMPLICATIONS: An observational study was conducted to compare various outcomes before and after the introduction of outpatient preoperative evaluation (OPE). Although smaller than anticipated, OPE for potential inpatients leads to a significant reduction of canceled cases and of length of admission. PMID- 11867391 TI - Cerebral hyperemia during recovery from general anesthesia in neurosurgical patients. AB - Changes in the cerebral circulation during recovery from neurosurgical anesthesia are poorly understood. We used transcranial Doppler to compare cerebral blood flow velocity changes (Vmca) during recovery after anesthesia. In the first part of the study, 30 patients were randomized to propofol- or isoflurane-based anesthesia. Vmca, mean arterial pressure (MAP), and CO(2) partial pressure (PaCO(2)) were measured before anesthesia, at tracheal extubation, at 5 to 60 min after extubation, and at 24 h after anesthesia. There was a 60% increase in Vmca above the awake value at extubation. The increase in Vmca was significant at least for 30 min after extubation. There was no difference between the Propofol and Isoflurane anesthesia groups. There was no correlation between Vmca and MAP or PaCO(2) at any time. In the second part of the study, Vmca, MAP, and jugular venous bulb saturation in oxygen (SjvO(2)) were measured after isoflurane anesthesia. SjvO(2) increased significantly at extubation, consistent with cerebral hyperemia. In conclusion, cerebral hyperemia occurs during recovery from general anesthesia independently of the anesthetic technique or hemodynamic or ventilatory changes. It is speculated that cerebral hyperemia is a nonspecific response to stress during emergence from anesthesia. IMPLICATIONS: Cerebral hyperemia occurs during emergence from general anesthesia. It might be one mechanism of cerebral complications in the early postoperative period. PMID- 11867392 TI - The effects of thiopental and propofol on cell swelling induced by oxygen/glucose deprivation in the CA1 pyramidal cell layer of rat hippocampal slices. AB - Cellular swelling has been implicated as an early process after cerebral ischemia. We compared the effects of two commonly used IV anesthetics, thiopental and propofol, on hippocampal CA1 pyramidal cell swelling induced by oxygen/glucose deprivation (OGD) in vitro. Experiments were performed in rat hippocampal slices. Cell swelling in the CA1 pyramidal cell layer was evaluated by determining light transmittance (LT) change through the slices and by histopathological examination. For LT experiments, OGD was induced for 10 min by superfusing slices with glucose-free artificial cerebrospinal fluid equilibrated with 95% nitrogen and 5% CO(2). Thiopental and propofol were present 10 min before and during the period of OGD. The results showed that thiopental (100 and 400 microM), but not propofol (40 and 160 microM), significantly prolonged latency to the peak of LT increase after the onset of OGD. Consistent with the LT experiments, histopathological examination revealed that thiopental, but not propofol, attenuated CA1 pyramidal cell expansion and the gap diminution between CA1 pyramidal cells induced by OGD. These results suggest that thiopental, but not propofol, reduces the neuronal cell swelling caused by OGD. Whether the reduction of cell swelling is related to reduction in cell injury caused by OGD remains to be investigated. IMPLICATIONS: We demonstrated that thiopental, but not propofol, attenuates ischemic neuronal swelling induced by oxygen/glucose deprivation in an in vitro ischemic model. PMID- 11867393 TI - Cerebral blood volume and blood flow responses to hyperventilation in brain tumors during isoflurane or propofol anesthesia. AB - Using computerized tomography, we measured absolute cerebral blood flow (CBF) and cerebral blood volume (CBV) in tumor, peri-tumor, and contralateral normal regions, at normocapnia and hypocapnia, in 16 rabbits with brain tumors (VX2 carcinoma), under isoflurane or propofol anesthesia. In both anesthetic groups, CBV and CBF were highest in the tumor region and lowest in the contralateral normal tissue. For isoflurane, a significant decrease in both CBV and CBF was observed in all tissue regions with hyperventilation (P < 0.05), but without accompanying changes in intracranial pressure. However, the percent reduction in regional CBF with hypocapnia was two times larger than that observed in the CBV response (P < 0.01). In contrast, there were no significant changes in CBV and CBF in the Propofol group with hyperventilation for all regions (P > 0.10). In addition, there were no differences between CBV values for isoflurane at hypocapnia when compared with CBV values for propofol at normo- or hypocapnia (P > 0.34 and P > 0.35, respectively, in the tumor regions). Our results indicate that propofol increases cerebral vascular tone in both neoplastic and normal tissue vessels compared with isoflurane. CBV and CBF during normocapnia were significantly greater in all regions (tumor, peri-tumor, and contralateral normal tissue) with isoflurane than with propofol. CBV and CBF remained responsive to hyperventilation only with isoflurane. IMPLICATIONS: In rabbits with brain tumors, brain blood flow and volume were significantly larger in all regions (tumor, peri-tumor, and contralateral normal tissue) with isoflurane than with propofol during normocapnia, and remained responsive to a reduction in PaCO(2). Consequently, during hypocapnia, brain blood flow and volume values with isoflurane were similar to values with propofol. PMID- 11867394 TI - A comparison of two tracheal intubation techniques with Trachlight and Fastrach in patients with cervical spine disorders. AB - Optimal airway management strategies in patients with an unstable cervical spine remain controversial. A newly designed lightwand device (Trachlight) or an intubating laryngeal mask (Fastrach) may avoid hyperextension of the neck. However, there are few objective data that guide us in selecting the appropriate devices. We conducted a prospective randomized study in 148 patients who received general anesthesia for whom the operations were related to the clinical and/or radiographic evidence of cervical abnormality. Trachlight or Fastrach was used for tracheal intubation with the head and neck held in a neutral position. In the Trachlight group, intubation was successful at the first attempt in 67 of 74 (90.5%) cases and at the second attempt in 5 (6.8%) cases. In contrast, in the Fastrach group, 54 of 74 (73.0%) patients were intubated within our protocol. The mean time for successful tracheal intubation at the first attempt was significantly shorter in the Trachlight group than in the Fastrach group. The Trachlight may be more advantageous for orotracheal intubation in patients with cervical spine disorders than the Fastrach with respect to reliability, rapidity, and safety. IMPLICATIONS: The Trachlight may be more advantageous for orotracheal intubation in patients with cervical spine disorders than the Fastrach with respect to reliability, rapidity and safety. PMID- 11867395 TI - Extravasation injury of the upper extremity by intravenous phenytoin. AB - IMPLICATIONS: Extravasation or IV injury attributable to phenytoin can cause severe soft tissue damage. Anesthesiologists are likely to encounter this problem and should recognize it and the potential complications. With increased awareness, the occurrence may be minimized. PMID- 11867396 TI - The primary action of epidural fentanyl after cesarean delivery is via a spinal mechanism. AB - We tested the hypotheses that the primary mechanism of action of epidural fentanyl after cesarean delivery is spinal and that very small dose epidural bupivacaine with epinephrine enhances this effect. After elective cesarean delivery, 100 parturients were randomized in a double-blinded design to four groups. Group I and II patients received a continuous 12 mL/h epidural infusion of bupivacaine 0.015% with epinephrine 1 microg/mL for 48 h and Groups III and IV received a 12 mL/h saline epidural infusion instead. Fentanyl 20 microg/mL was administered via a patient-controlled analgesia device either into the epidural infusion (Groups I and IV) or IV (Groups II and III). When compared to patients receiving epidural fentanyl, those receiving IV fentanyl required larger mean infused and total dose of fentanyl (P < 0.0001), reported more pain (P < 0.001), and had a more frequent incidence of excessive sedation (P < 0.01), nausea (P < 0.01), and vomiting (P < 0.01). Plasma concentrations of fentanyl were larger for Group II and III than for Groups I and IV (P < 0.001) at 24 and 48 h. Our results support the hypothesis that the primary mechanism of analgesia of epidural fentanyl after cesarean delivery is spinal. Our data also show that the total required dose of epidural, but not IV, fentanyl is reduced by very small dose epidural bupivacaine and epinephrine (Group I versus Group IV, P < 0.02 and Group II vs Group III, not significant). IMPLICATIONS: Fentanyl administered epidurally to parturients after cesarean delivery has a primarily spinal mechanism of action and this effect is enhanced by very small dose epidural bupivacaine and epinephrine. PMID- 11867397 TI - Spinal ropivacaine for cesarean delivery: a comparison of hyperbaric and plain solutions. AB - We compared, in this prospective, randomized, double-blinded study, the characteristics of spinal anesthesia with plain and hyperbaric ropivacaine for elective cesarean delivery. We hypothesized that the addition of glucose would change the onset, offset, and extent of motor and sensory block from intrathecal ropivacaine. Forty ASA physical status I--II women were given 25 mg of either ropivacaine (n = 20) or ropivacaine in 8.3% glucose (n = 20) intrathecally, via a combined spinal/epidural technique in the right lateral position. Sensory changes to ice and pinprick and motor block (Bromage score) were recorded at 2.5-min intervals. Adequate anesthesia for surgery was achieved in all patients in the Hyperbaric group, whereas in the Plain group, five (25%) patients required epidural top-up because of insufficient rostral spread (P < 0.05). With hyperbaric ropivacaine, we found the following: higher cephalic spread (median [range] maximum block height to pinprick, T1 [T4 to C2] versus T3 [T11 to C3], P < 0.001); lower coefficient of variation of maximum block height (17.7% vs 21.9%); faster onset to T4 dermatome (mean [SD] 7.7 [4.9] vs 16.4 [14.1] min, P = 0.015); and faster recovery to L1 (189.0 [29.6] vs 215.5 [27.0] min, P = 0.01). The onset of complete motor block (9.9 [5.3] vs 13.8 [5.4] min, P = 0.027) and complete recovery (144.8 [28.4] vs 218.5 [56.8] min, P < 0.001) was also faster. No neurologic symptoms were found at 24 h. IMPLICATIONS: We compared hyperbaric and plain ropivacaine for combined spinal/epidural analgesia in the lateral position in patients undergoing elective cesarean delivery. Hyperbaric ropivacaine produced more rapid block with faster recovery and less requirement for epidural supplementation compared with plain ropivacaine. PMID- 11867398 TI - Prolonged neuromuscular block after rocuronium in postpartum patients. AB - Postpartum patients have not completely lost the weight gained during pregnancy. Drug dosing according to total body weight (TBW) can cause exaggerated effects and dosing by lean body mass (LBM) may provide a more consistent response despite the increased weight. We compared the duration of a rocuronium neuromuscular block in 22 women undergoing postpartum tubal ligation 31--79 h after delivery, with that in 22 women undergoing gynecological surgery. Anesthesia was induced and maintained with propofol and alfentanil. Half of the patients in each of the Postpartum and Control groups received a bolus dose of rocuronium 0.6 mg/kg TBW and the remaining half received rocuronium 0.6 mg/kg LBM. Neuromuscular block was monitored by electromyography and the ulnar nerve was stimulated transcutaneously using a train-of-four pattern. When rocuronium was given by TBW, median (range) duration of neuromuscular block until 25% recovery of the first twitch response was longer in the Postpartum group, 35.3 (29.7--48.7) min, compared with the Control group, 24.8 (21.5--28.6) min (P < 0.001). After dosing by LBM, the duration of block was similar between groups. The prolonged block with rocuronium in the Postpartum patients can be explained by relative drug overdose when dose calculation is based on their temporarily increased body weight. IMPLICATIONS: Neuromuscular block is prolonged in the postpartum period after standard doses of rocuronium. Drug administration according to lean body mass will produce a more consistent duration of block. PMID- 11867399 TI - Herbal medicine use in parturients. AB - Alternative medicine use has increased dramatically over the last decade. Recently a 22% incidence of herbal medicine use in presurgical patients was reported. Of concern is the potential for these medications to cause adverse drug herb interactions or other effects such as bleeding complications. We sought to determine the prevalence and pattern of use of herbal remedies in parturients. A one-page questionnaire examining the use of all prescription and nonprescription medications, including herbal remedies, was sent to parturients expected to deliver within 20 wk who had preregistered with the hospital's admissions office. Sixty-one percent of the parturients responded to the survey, with 7.1% of parturients reporting the use of herbal remedies. Only 14.6% of users considered them to be medications. Parturients in the 41--50 yr age bracket (5.6% of parturients) were the most likely to use herbal remedies (17.1% rate of use in this age group). Many parturients who took herbal remedies (46%) did so on the recommendation of their health care provider. IMPLICATIONS: Herbal medicine use may be less prevalent in the parturient population, however, some health care providers may be promoting their use during pregnancy. PMID- 11867400 TI - Spinal cord stimulation in postherpetic neuralgia and in acute herpes zoster pain. AB - We studied the effects of spinal cord stimulation (SCS) on postherpetic neuralgia (PHN). Data of 28 patients were prospectively investigated over a median period of 29 (quartiles 9--39) mo. In addition, four patients with acute herpes zoster (HZ) pain were studied simultaneously. After intractable pain for more than 2 yr, long-term pain relief was achieved in 23 (82%) PHN patients (median, 70 yr) during SCS treatment confirmed by a median decrease from 9 to 1 on the visual analog scale (P < 0.001). In five cases with serious comorbidity, the initial pain alleviation could not be stabilized. Spontaneous improvement was always confirmed or excluded by SCS inactivation tests at quarterly intervals. Eight patients discontinued SCS permanently because of complete pain relief after stimulation periods of 3--66 mo, whereas 2 reestablished SCS because of recrudescence after 2 and 6 mo. Considerable impairments in everyday life, objectified by the pain disability index, were also significantly improved (P < 0.001). In 4 patients with acute HZ pain, SCS was promptly effective and after periods of 2.5 (quartiles 2--3) months the pain had subsided. SCS seems to offer a therapeutic option for pharmacological nonresponders. IMPLICATIONS: In many patients with postherpetic neuralgia and acute herpes zoster pain is not satisfactorily alleviated with pharmacological approaches. We report on 23 of 28 patients with postherpetic neuralgia and 4 of 4 with acute herpes zoster whose chronic pain was improved by electrical spinal cord stimulation. PMID- 11867401 TI - Apraclonidine attenuates the increases in spinal excitatory amino acid release in rats with adjuvant-induced inflammation. AB - The release of excitatory amino acids (EAAs), nitric oxide, and prostaglandins plays a critical role in the development of peripheral tactile and thermal hypersensitivity after the induction of knee joint inflammation. In this study, we used a model of chronic spinal microdialysis to examine the effect of complete Freund's adjuvant (CFA)-induced inflammation on the spinal release of EAAs and also assessed the antinociceptive effect of a new alpha(2)-adrenergic agonist, apraclonidine, by using this model. Male Sprague-Dawley rats were implanted with microdialysis catheters. CFA was injected into the plantar surface of the left hindpaw to induce inflammation. Concentrations of amino acids in dialysate and thermal and tactile withdrawal latency were evaluated for 1 wk. Intraplantar injection of CFA evoked a significant release of glutamate, aspartate, and citrulline for 6 days. Three milligrams of intraperitoneal apraclonidine significantly suppressed the release of EAAs and citrulline. Apraclonidine was given intraperitoneally 2--3 days after CFA injection. Prominent thermal and tactile allodynia was observed for 6 days. Our results show that the significant modulatory effect of the alpha(2)-adrenergic agonist apraclonidine on the release of EAAs may account for its antinociceptive properties in adjuvant-induced inflammation. IMPLICATIONS: This study showed a novel finding that the hypersensitivity state seems to be dependent on increased release of spinal excitatory amino acids (EAAs), and the significant modulatory effect of the alpha(2)-adrenergic agonist apraclonidine on the release of spinal EAAs accounts for its analgesic properties in adjuvant-induced inflammation. PMID- 11867402 TI - Ultrasound guidance for the psoas compartment block: an imaging study. AB - We conducted this study to develop an ultrasound-guided approach to the psoas compartment and to assess its feasibility and accuracy by means of computed tomography (CT). Two examiners performed ultrasound-guided approaches at three levels (L2-3, L3-4, and L4-5) on 10 embalmed cadavers, which were seated prone. After each needle had been advanced into the psoas compartment under ultrasound guidance, the positions of their tips were computed by using two coordinates (A and B). Subsequently, axial transverse CT scans were made to verify the ultrasound measurements by using the same coordinates. In total, 48 approaches were performed (Examiner 1, n = 20; Examiner 2, n = 28). CT revealed that 47 of 48 ultrasound-guided approaches were performed exactly. In 1 of 48 approaches (L3 4), the tip of the needle was located posterior to the psoas muscle. The median differences between ultrasound and CT coordinates were 0.3 plus minus 0.3 cm for A and 0.2 plus minus 0.3 for B. Kendall's coefficient of concordance was 0.9 (P < 0.001) between ultrasound and CT measurements for both coordinates. These results indicate that ultrasound enables exact needle placement, as proved by CT. We conclude that ultrasound guidance might be a useful adjunct to increase the safety and efficacy of the psoas compartment block at these levels. IMPLICATIONS: We developed an ultrasound-guided approach to the psoas compartment at the levels L2-3, L3-4, and L4-5. Feasibility and accuracy were tested on embalmed cadavers and verified by means of computed tomography. Ultrasound guidance proved to be feasible and accurate for the performance of psoas compartment blocks. PMID- 11867403 TI - The antinociceptive and histologic effect of sciatic nerve blocks with 5% butamben suspension in rats. AB - Butamben, a lipophilic local anesthetic of the ester class, produces a differential nerve block of long duration. Epidural and peripheral nerve blocks with butamben, formulated as a 5%--10% suspension, result in prolonged analgesia without significant motor blockade. We evaluated the effect of butamben sciatic nerve block on antinociception using the rat paw formalin test, as well as withdrawal latencies to thermal stimulation, and assessed histologic changes in the nerve. After right sciatic nerve block with butamben 5% or saline, responses to intradermal injection of 5% formalin were recorded in randomly selected groups of 6 animals each on days 1, 2, 5, 10, 21, and 28. In an additional group of 8 thermal challenges to both hind paws were recorded at 1, 2, 5, 7, 10, 14, 17, 21, and 28 days after right sciatic butamben 5% blocks. Butamben injection decreased the formalin-induced flinches on days 2, 5, 10, 21 and 28 and decreased thermal challenges on days 1 through 17. Histologic changes were minimal. This study demonstrates a prolonged antinociceptive effect from butamben nerve block to both formalin-induced nociception and heat hyperalgesia, without an effect on gross motor function or histologic morphology. IMPLICATIONS: Butamben 5% nerve blocks produced a prolonged antinociceptive effect to formalin-induced nociception and heat hyperalgesia, without significant motor effect or evidence of substantial histologic changes. PMID- 11867404 TI - The efficacy of hemodynamic and T-wave criteria for detecting intravascular injection of epinephrine test dose in propofol-anesthetized adults. AB - A recent study demonstrated 100% effectiveness of hemodynamic criteria during propofol anesthesia, when a full dose of an epinephrine (15 microg)-containing test dose was injected intravascularly. We designed this dose-response study to determine minimal effective epinephrine doses and efficacies of hemodynamic and T wave criteria for detecting intravascular injection of the epinephrine test dose in propofol-anesthetized adults. Eighty healthy adult patients were randomly assigned to one of four groups according to a simulated IV test dose using propofol (133 microg center dot kg(-1) center dot min(-1)) and nitrous oxide (FIO(2) = 0.33) anesthesia after endotracheal intubation (n = 20 each). The Saline group received 3 mL of normal saline IV; the Epinephrine-15 group received 3 mL of 1.5% lidocaine containing 15 microg epinephrine; and the Epinephrine-10 and -5 groups received 2 and 1 mL of the test dose of the identical components, respectively. Heart rate (HR), systolic blood pressure (SBP), and lead II of the electrocardiogram were recorded continuously for 5 min after the IV injection of the study drug via a peripheral vein. Sensitivities and specificities of 100% were obtained based on the modified HR (positive if greater-than-or-equal to 10 bpm increase) and the T-wave (positive if greater-than-or-equal 25% in amplitude) criteria if greater-than-or-equal 5 microg of epinephrine was injected IV. Based on the SBP criterion (positive if greater-than-or-equal 15 mm Hg increase), however, 100% sensitivity and specificity were associated only with greater-than or-equal 10 microg of epinephrine doses. These results suggest that the minimal effective epinephrine doses for detecting unintentional intravascular injection are 5 microg based on the HR and T-wave criteria, and 10 microg based on the SBP criterion in adult patients anesthetized with propofol and nitrous oxide. IMPLICATIONS: Accidental migration of an epidural catheter into a blood vessel is often detected by hemodynamic changes after injecting an epidural test dose containing epinephrine. Our results suggest that 5 microg of epinephrine is not adequate to reliably produce hemodynamic and T-wave alterations in adult patients during propofol anesthesia. PMID- 11867405 TI - Assessing sedation with regional anesthesia: inter-rater agreement on a modified Wilson sedation scale. AB - A valid and reliable means for measuring sedation during regional anesthesia would be valuable for both research and practice. Current methods of monitoring sedation include machine-, patient-, and observer-based assessment. The reliability of machine-based methods is limited at lower levels of sedation, whereas patient-based methods are impractical at higher levels. Observer-based methods offer the best alternative for assessing sedation during regional anesthesia; however, their reliability has not been adequately documented. We examined the interrater reliability of the Wilson sedation scale. Sedation was assessed by pairs of anesthesia care providers in 100 patients undergoing surgical procedures with regional anesthesia. On the basis of the findings, the scale was modified, and 50 additional patients were assessed. The study protocol called for a series of standardized stimuli administered by a research assistant. Raters were blinded to each other's ratings. Interrater reliability was assessed by using the kappa statistic, a measure of actual agreement beyond agreement by chance. When continuing checks on its operationalization and reliability are included, the modified Wilson scale provides a simple and reliable means by which to assess and monitor intraoperative sedation. IMPLICATIONS: We evaluated the interrater reliability of the Wilson scale for measuring sedation during regional anesthesia. Paired anesthesia care providers' ratings of patient sedation indicated very good interrater reliability in both the original scale and a modified version. The modified Wilson scale provides a quick noninvasive means of monitoring sedation during regional anesthesia. PMID- 11867406 TI - Bacterial growth in ropivacaine hydrochloride. AB - IMPLICATIONS: Drugs affecting bacterial growth may influence the occurrence of postoperative infections. Ropivacaine 10 mg/mL killed Staphylococcus aureus and Escherichia coli; ropivacaine 2 mg/mL supported the growth of E. coli. PMID- 11867407 TI - Morbid obesity and tracheal intubation. AB - The tracheas of obese patients may be more difficult to intubate than those of normal-weight patients. We studied 100 morbidly obese patients (body mass index >40 kg/m(2)) to identify which factors complicate direct laryngoscopy and tracheal intubation. Preoperative measurements (height, weight, neck circumference, width of mouth opening, sternomental distance, and thyromental distance) and Mallampati score were recorded. The view during direct laryngoscopy was graded, and the number of attempts at tracheal intubation was recorded. Neither absolute obesity nor body mass index was associated with intubation difficulties. Large neck circumference and high Mallampati score were the only predictors of potential intubation problems. Because in all but one patient the trachea was intubated successfully by direct laryngoscopy, the neck circumference that requires an intervention such as fiberoptic bronchoscopy to establish an airway remains unknown. We conclude that obesity alone is not predictive of tracheal intubation difficulties. IMPLICATIONS: In 100 morbidly obese patients, neither obesity nor body mass index predicted problems with tracheal intubation. However, a high Mallampati score (greater-than-or-equal to 3) and large neck circumference may increase the potential for difficult laryngoscopy and intubation. PMID- 11867408 TI - The Laryngeal Mask Airway ProSeal(TM) as a temporary ventilatory device in grossly and morbidly obese patients before laryngoscope-guided tracheal intubation. AB - We determined the efficacy of the laryngeal mask airway ProSeal(TM) (PLMA) as a temporary ventilatory device in morbidly obese patients before laryngoscope guided tracheal intubation. Sixty patients (body mass index 35--60 kg/m(2)) scheduled for elective surgery, who preferred airway management under general anesthesia, were studied. The induction of anesthesia was with midazolam/fentanyl/propofol and maintenance was with sevoflurane 1%--3% in oxygen 100%. The PLMA was inserted and an effective airway established. Rocuronium was given IV for paralysis. Oropharyngeal leak pressure, ease of gastric tube placement, residual gastric volume, fiberoptic position of the airway/drainage tube, and ease of ventilation at a tidal volume of 8 mL/kg was determined. The PLMA was then removed and laryngoscope-guided tracheal intubation attempted. The number of insertion/intubation attempts (maximum two each) and time taken to establish an effective airway with each device were recorded. An effective airway was obtained at the first insertion attempt in 90% of patients (54/60) and at the second attempt in 10% (6/60). The time taken to provide an effective airway was 15 plus minus 7 s (7--42 s). Oropharyngeal leak pressure was 32 plus minus 8 cm H(2)O (12--40 cm H(2)O). The residual gastric volume was 36 plus minus 46 mL (0- 240 mL). Positive pressure ventilation without air leaks was possible in 95% of patients (57/60). The vocal cords were seen from the airway tube in 75% of patients (45/60), but the esophagus was not seen. The fiberoptic view from the drainage tube revealed mucosa in 93% of patients (56/60) and an open upper esophageal sphincter in 7% (4/60). Tracheal intubation was successful at the first attempt in 90% of patients (54/60), at the second attempt in 7% (4/60), and failed in 3% (2/60). In these latter two patients, the PLMA was reinserted and surgery performed uneventfully with the PLMA. The time taken to tracheally intubate the patient was 13 plus minus 10 s (8--51 s). There were no episodes of hypoxia (SpO(2) <90%) or other adverse events. There were no differences in insertion success rate, or the time to successful insertion between the PLMA and laryngoscope-guided intubation. We conclude that the PLMA is an effective temporary ventilatory device in grossly or morbidly obese patients before laryngoscope-guided tracheal intubation. IMPLICATIONS: The laryngeal mask airway ProSeal(TM) is an effective temporary ventilatory device in grossly and morbidly obese patients before laryngoscope-guided tracheal intubation. PMID- 11867409 TI - The effects of abdominal opening on respiratory mechanics during general anesthesia in normal and morbidly obese patients: a comparative study. AB - Morbid obesity has a profound effect on respiratory mechanics and gas exchange. However, most studies were performed in morbidly obese patients before or after anesthesia. We tested the hypothesis that anesthesia and abdominal opening could modify the elastic and resistive properties of the respiratory system. Eleven morbidly obese and eight normal-weight patients scheduled for gastric binding and cancer treatment, respectively, under laparotomy were studied. Respiratory mechanics, partitioned into its lung and chest wall components, were investigated during surgery by means of the end-inspiratory inflation occlusion method and esophageal balloon at five time points. Static respiratory and lung compliance were markedly reduced in obese patients; on the contrary, static compliance of chest wall presented comparable values in both groups. Obese patients also presented higher resistances of the total respiratory system, lung and chest wall, as well as "additional" lung resistance. Mainly in obese patients, laparotomy provoked a significant increase in lung compliance and decrease in "additional" lung resistance 1 h after the peritoneum was opened, which returned to original values after the peritoneum had been closed (P < 0.005). In obese patients, low respiratory compliance and higher airway resistance were mainly determined by the lung component. PMID- 11867410 TI - The prophylactic effect of tropisetron on epidural morphine-related nausea and vomiting: a comparison of dexamethasone with saline. AB - Tropisetron is a 5-hydroxytryptamine subtype 3 receptor antagonist that is primarily used in the prevention of chemotherapy-induced nausea and vomiting. We evaluated the prophylactic effect of tropisetron on postoperative nausea and vomiting associated with epidural morphine. Dexamethasone and saline served as controls. One-hundred twenty women (n = 40 in each of three groups) undergoing abdominal total hysterectomy under epidural anesthesia were enrolled in this randomized, double-blinded, and placebo-controlled study. At the end of surgery, Group 1 received IV tropisetron 5 mg, whereas Groups 2 and 3 received dexamethasone 5 mg and saline, respectively. We found that tropisetron did not significantly reduce the occurrence of nausea and vomiting associated with epidural morphine. Dexamethasone, however, reduced the total incidence of nausea and vomiting from 59% to 21% (P < 0.01) and the percentage of patients requiring rescue antiemetic from 38% to 13% (P < 0.05). We conclude that IV tropisetron 5 mg did not prevent the occurrence of postoperative nausea and vomiting associated with epidural morphine. IV dexamethasone 5 mg was effective for this purpose. IMPLICATIONS: We compared the prophylactic IV administration of tropisetron 5 mg to prevent postoperative nausea and vomiting (PONV) associated with epidural morphine with dexamethasone 5 mg and saline in women undergoing hysterectomy. We found that tropisetron 5 mg did not significantly reduce the occurrence of PONV associated with epidural morphine. Dexamethasone 5 mg was effective for this purpose. PMID- 11867411 TI - Latex allergy desensitization by exposure protocol: five case reports. AB - Two clinical reports dealing with subcutaneous desensitization to latex are available, but these treatments were reported to have significant side effects. To investigate an alternative method for latex desensitization, we present five cases of immunoglobulin E-mediated latex allergy in which a specific desensitization was successfully performed by means of an original exposure protocol. Five female patients with proven latex allergy underwent an original contact desensitization by wearing latex gloves daily, progressively increasing exposure to latex. All patients reached a final latex exposure time of 60 min in both hands. The desensitization protocol was completely successful in all patients. Twelve months after the beginning of the desensitization program, all the treated subjects were able to wear latex gloves daily without any clinical manifestations. This study provides evidence that a successful contact desensitization to latex is possible with no side effects. The adopted exposure protocol seems to be completely safe and effective. However, further studies in larger groups of subjects are necessary to fully confirm these preliminary results. IMPLICATIONS: We present five patients with latex allergy who underwent desensitization by a new cutaneous exposure protocol. This study provides evidence that a safe therapeutic approach to latex allergy is possible. PMID- 11867412 TI - A streamlined pharynx airway liner: a pilot study in 22 patients in controlled and spontaneous ventilation. AB - IMPLICATIONS: An inexpensive, single-use alternative device to the laryngeal mask and cuffed oropharyngeal airway appears useful in controlled and spontaneous ventilation. The hollow soft plastic airway may act as a sump for pooled secretions, possibly minimizing regurgitation risks. Without a cuff, important cost savings are likely. PMID- 11867413 TI - Gastroesophageal reflux and aspiration of gastric contents. PMID- 11867414 TI - Place the bubble solution with your fingertip. PMID- 11867415 TI - Clinical productivity metrics. PMID- 11867416 TI - Intrapleural--another approach to sensory phrenic nerve block. PMID- 11867417 TI - Epidural catheter placement using electrical stimulation test. PMID- 11867418 TI - The second gas effect is not statistically valid. PMID- 11867419 TI - Continuous patient oxygenation during endotracheal intubation through the LMA Fastrack. PMID- 11867421 TI - Your Journal, Our Journal. PMID- 11867420 TI - Withdrawal of antihypertensive drugs before anesthesia. PMID- 11867422 TI - Enhancing Continuity of Care: Residency Training in an Integrated Inpatient- Partial Hospital Program. AB - This paper describes a teaching/service model that integrates inpatient, partial hospital, and intensive outpatient treatment. In this model, individual multidisciplinary treatment teams retain responsibility for a patient's care for any or all of three levels of intensity of services accessed during an episode of illness. This teaching/service model allows residents to follow patients for an average of 2.5 weeks across an entire acute episode of care compared with the 7.3 days of the average inpatient stay at the inpatient facility. The opportunity to continue treatment in step-down settings over longer periods of time allows residents and medical students to develop a fuller understanding of their patients. The authors believe that this continuum-care service model more efficiently trains residents in multiple aspects of psychiatric practice and provides patients with better care than the traditional inpatient-care service model. PMID- 11867423 TI - Residents' Morning Report in Psychiatry Training: Description of a Model and a Survey of Resident Attitudes. AB - A residents' morning report, adapted from the traditional internal medicine model, was introduced into a psychiatry residency curriculum to supplement bedside teaching for junior residents during inpatient service rotations by focusing on case presentations, clinical reasoning, and evidence-based decision making skills. This paper describes the design of the report and findings from surveys of residents who participated in the report over a 3-year period. The surveys demonstrate high levels of satisfaction with the exercise and offer insight into factors that may optimize or impede learning in the morning report setting in psychiatry. PMID- 11867424 TI - Family Evaluation Clinic: Training Psychiatrists to Think Systemically. AB - A family evaluation training program, Family Evaluation Clinic (FEC), was conducted in an outpatient child psychiatry clinic serving a low-income, African American, urban population. To determine the success of the program in meeting its training goals, the authors reviewed patient attendance rates and collected survey data from trainees who participated in evaluations conducted between 1994 and 2000. Patient attendance rates were high, and trainees indicated that the program was helpful and valuable to them in their current practice. The authors discuss advantages and disadvantages of FEC as a training method and share recommendations for future family evaluation training in a general residency program. PMID- 11867425 TI - The Challenge of Empathy: A Pilot Study of the Use of Standardized Patients to Teach Introductory Psychopathology to Medical Students. AB - This pilot study compares standardized and actual psychiatric patients used to teach introductory psychopathology. In a blinded manner, students and psychiatric faculty interviewed both types of patients, using a small group format. Before and after the course, students and faculty completed a questionnaire about expectations and effectiveness. Students were divided in their opinions of standardized patients but generally preferred actual patients. Faculty were initially noncommittal, but after the course they strongly preferred actual patients. Although standardized patients offer some advantages over actual patients, limitations included difficulty developing empathy. Standardized patients could potentially be trained to convey emotions realistically, but further study of this is needed. PMID- 11867426 TI - Standards for Psychiatry Clerkship Directors. AB - The authors review the literature relevant to the position of Psychiatry Clerkship Director (PCD) and propose standards regarding the expectations for this position. The standards address qualifications, duties, and competencies in the areas of leadership, administration, education, mentoring, and scholarship, as well as the resources of time, administrative assistance, budget, and compensation required to carry out these duties. This paper has been endorsed by the Council of the Association of Directors of Medical Student Education in Psychiatry (ADMSEP), by the American Psychiatric Association's Committee on Medical Student Education, and by the Executive Committee of the American Association of Chairmen of Departments of Psychiatry. PMID- 11867427 TI - Lost in the DSM-IV Checklist : Empathy, Meaning, and the Doctor--Patient Relationship. PMID- 11867428 TI - That's My Doctor. PMID- 11867430 TI - Rebellious Rhapsody: Metal, Rap, Community, and Individuation. AB - Music can be a powerful force and tool in the life of an adolescent. It forms a social context and informs the adolescent about the adult world through the lens of artists' lives, language, and presence as models. Allegiance to a form of music is allegiance to those who make it, a way to friendship and kinship, and a road to personal identity through belonging. In their relationships formed through music, teens can create a sense of community that may be lacking in the life of family. The rebellious music of earlier generations has given rise to complex musical genres, rap and heavy metal, that are strong in defiance and controversial in their violent and sexual content. What do these musical affiliations tell us about certain segments of adolescent development and culture? The authors consider this question by exploring the form and content of the music while using it to illuminate psychodynamic and psychosocial aspects of adolescent development. PMID- 11867431 TI - Chromatophore vesicles of Rhodobacter capsulatus contain on average one F(O)F(1) ATP synthase each. AB - ATP synthase is a unique rotary machine that uses the transmembrane electrochemical potential difference of proton (Delta(H(+))) to synthesize ATP from ADP and inorganic phosphate. Charge translocation by the enzyme can be most conveniently followed in chromatophores of phototrophic bacteria (vesicles derived from invaginations of the cytoplasmic membrane). Excitation of chromatophores by a short flash of light generates a step of the proton-motive force, and the charge transfer, which is coupled to ATP synthesis, can be spectrophotometrically monitored by electrochromic absorption transients of intrinsic carotenoids in the coupling membrane. We assessed the average number of functional enzyme molecules per chromatophore vesicle. Kinetic analysis of the electrochromic transients plus/minus specific ATP synthase inhibitors (efrapeptin and venturicidin) showed that the extent of the enzyme-related proton transfer dropped as a function of the inhibitor concentration, whereas the time constant of the proton transfer changed only marginally. Statistical analysis of the kinetic data revealed that the average number of proton-conducting F(O)F(1) molecules per chromatophore was approximately one. Thereby chromatophores of Rhodobacter capsulatus provide a system where the coupling of proton transfer to ATP synthesis can be studied in a single enzyme/single vesicle mode. PMID- 11867432 TI - New stochastic strategy to analyze helix folding. AB - We propose an alternative stochastic strategy to search secondary structures based on the generalized simulated annealing (GSA) algorithm, by using conformational preferences based on the Ramachandran map. We optimize the search for polypeptide conformational space and apply to peptides considered to be good alpha-helix promoters above a critical number of residues. Our strategy to obtain conformational energies consist in coupling a classical force field (THOR package) with the GSA procedure, biasing the Phi x Psi backbone angles to the allowed regions in the Ramachandran map. For polyalanines we obtained stable alpha-helix structures when the number of residues were equal or exceeded 13 amino acids residues. We also observed that the energy gap between the global minimum and the first local minimum tends to increase with the polypeptide size. These conformations were generated by performing 2880 stochastic molecular optimizations with a continuum medium approach. When compared with molecular dynamics or Monte Carlo methods, GSA can be considered the fastest. PMID- 11867433 TI - Molecular mechanisms of calcium and magnesium binding to parvalbumin. AB - Molecular dynamics simulations have been used to investigate the relationship between the coordinating residues of the EF-hand calcium binding loop of parvalbumin and the overall plasticity and flexibility of the protein. The first simulation modeled the transition from Ca(2+) to Mg(2+) coordination by varying the van der Waals parameters for the bound metal ions. The glutamate at position 12 could be accurately and reversibly seen to be a source of selective bidentate ligation of Ca(2+) in the simulations. A second simulation correlated well with the experimental observation that an E101D substitution at EF loop position 12 results in a dramatically less tightly bound monodentate Ca(2+) coordination by aspartate. A final set of simulations investigated Ca(2+) binding in the E101D mutant loop in the presence of applied external forces designed to impose bidentate coordination. The results of these simulations illustrate that the aspartate is capable of attaining a suitable orientation for bidentate coordination, thus implying that it is the inherent rigidity of the loop that prevents bidentate coordination in the parvalbumin E101D mutant. PMID- 11867434 TI - Vibrational resonances in biological systems at microwave frequencies. AB - Many biological systems can be expected to exhibit resonance behavior involving the mechanical vibration of system elements. The natural frequencies of such resonances will, generally, be in the microwave frequency range. Some of these systems will be coupled to the electromagnetic field by the charge distributions they carry, thus admitting the possibility that microwave exposures may generate physiological effects in man and other species. However, such microwave excitable resonances are expected to be strongly damped by interaction with their aqueous biological environment. Although those dissipation mechanisms have been studied, the limitations on energy transfers that follow from the limited coupling of these resonances to the electromagnetic field have not generally been considered. We show that this coupling must generally be very small and thus the absorbed energy is so strongly limited that such resonances cannot affect biology significantly even if the systems are much less strongly damped than expected from basic dissipation models. PMID- 11867435 TI - A mechano-electrochemical model of radial deformation of the capillary glycocalyx. AB - A mechano-electrochemical theory of the surface glycocalyx on capillary endothelial cells is presented that models the structure as a mixture of electrostatically charged macromolecules hydrated in an electrolytic fluid. Disturbances arising from mechanical deformation are introduced as perturbations away from a nearly electroneutral equilibrium environment. Under mechanical compression of the layer, such as might occur on the passing of stiff leukocytes through capillaries, the model predicts that gradients in the electrochemical potential of the compressed layer cause a redistribution of mobile ions within the glycocalyx and a rehydration and restoration of the layer to its equilibrium dimensions. Because of the large deformations of the glycocalyx arising from passing leukocytes, nonlinear kinematics associated with finite deformations of the layer are accounted for in the theory. A pseudo-equilibrium approximation is invoked for the transport of the mobile ions that reduces the system of coupled nonlinear integro-differential equations to a single nonlinear partial differential equation that is solved numerically for the compression and recovery of the glycocalyx using a finite difference method on a fixed grid. A linearized model for small strains is also obtained as verification of the finite difference solution. Results of the asymptotic analysis agree well with the nonlinear solution in the limit of small deformations of the layer. Using existing experimental and theoretical estimates of glycocalyx properties, the glycocalyx fixed-charge density is estimated from the analysis to be approximately 1 mEq/l, i.e., we estimate that there exists approximately one fixed charge on the glycocalyx for every 100 ions in blood. Such a charge density would result in a voltage differential between the undeformed glycocalyx and the capillary lumen of approximately 0.1 mV. In addition to providing insight into the mechano electrochemical dynamics of the layer under deformation, the model suggests several methods for obtaining improved estimates of the glycocalyx fixed-charge density and permeability in vivo. PMID- 11867436 TI - Continuum and atomistic modeling of ion partitioning into a peptide nanotube. AB - Continuum and atomistic descriptions of the partitioning of ions into a self assembled (D,L)-octapeptide nanotube, cyclo[-(L-Ala-D-Ala)(4)-], are presented. Perturbation free energy calculations, including Ewald electrostatics, are used to estimate the electrostatic component of the excess free energy of charging Li(+), Na(+), Rb(+), and Cl(minus sign) ions inside the nanotube. The radial density and orientational distribution of water around the ion is calculated for the ion at two different positions inside the tube; it is seen that the calculated distributions are sensitive to the location of the ions. Two different continuum electrostatic models are formulated to describe the ion solvation inside the nanotube. When enhanced orientational structuring of water dipoles is evidenced, explicitly including the first solvation shell as part of the low dielectric nanotube environment provides good agreement with molecular dynamics simulations. When water orientational structuring is as in the reference bulk solvent, we find that treating the first shell water explicitly or as a high dielectric continuum leads to similar results. These results are discussed, and their importance for continuum electrostatic modeling of ion channels are highlighted. PMID- 11867438 TI - Molecular modeling and dynamics of the sodium channel inactivation gate. AB - The intracellular linker L(III-IV) of voltage-gated sodium channels is known to be involved in their mechanism of inactivation. Its primary sequence is well conserved in sodium channels from different tissues and species. However, the role of charged residues in this region, first thought to play an important role in inactivation, has not been well identified, whereas the IFM triad (I1488 M1490) has been characterized as the crucial element for inactivation. In this work, we constructed theoretical models and performed molecular dynamics simulations, exploring the role of L(III-IV)-charged residues in the presence of a polar/nonpolar planar interface represented by a dielectric discontinuity. From structural predictions, two alpha-helical segments are proposed. Moreover, from dynamics simulations, a time-conserved motif is detected and shown to play a relevant role in guiding the inactivation particle toward its receptor site. PMID- 11867437 TI - Structure and dynamics of zymogen human blood coagulation factor X. AB - The solution structure and dynamics of the human coagulation factor X (FX) have been investigated to understand the key structural elements in the zymogenic form that participates in the activation process. The model was constructed based on the 2.3-A-resolution x-ray crystallographic structure of active-site inhibited human FXa (PDB:1XKA). The missing gamma-carboxyglutamic acid (GLA) and part of epidermal growth factor 1 (EGF1) domains of the light chain were modeled based on the template of GLA-EGF1 domains of the tissue factor (TF)-bound FVIIa structure (PDB:1DAN). The activation peptide and other missing segments of FX were introduced using homology modeling. The full calcium-bound model of FX was subjected to 6.2 ns of molecular dynamics simulation in aqueous medium using the AMBER6.0 package. We observed significant reorientation of the serine-protease (SP) domain upon activation leading to a compact multi-domain structure. The solution structure of zymogen appears to be in a well-extended conformation with the distance between the calcium ions in the GLA domain and the catalytic residues estimated to be approximately 95 A in contrast to approximately 83 A in the activated form. The latter is in close agreement with fluorescence studies on FXa. The S1-specificity residues near the catalytic triad show significant differences between the zymogen and activated structures. PMID- 11867439 TI - Temperature dependence of protein dynamics: computer simulation analysis of neutron scattering properties. AB - The temperature dependence of the internal dynamics of an isolated protein, bovine pancreatic trypsin inhibitor, is examined using normal mode analysis and molecular dynamics (MD) simulation. It is found that the protein exhibits marked anharmonic dynamics at temperatures of approximately 100-120 K, as evidenced by departure of the MD-derived average mean square displacement from that of the harmonic model. This activation of anharmonic dynamics is at lower temperatures than previously detected in proteins and is found in the absence of solvent molecules. The simulation data are also used to investigate neutron scattering properties. The effects are determined of instrumental energy resolution and of approximations commonly used to extract mean square displacement data from elastic scattering experiments. Both the presence of a distribution of mean square displacements in the protein and the use of the Gaussian approximation to the dynamic structure factor lead to quantified underestimation of the mean square displacement obtained. PMID- 11867440 TI - Calculating the bulk modulus for a lipid bilayer with nonequilibrium molecular dynamics simulation. AB - Nonequilibrium molecular dynamics (NEMD) computer simulations are used to calculated the bulk modulus for a dimyristoylphosphatidylcholine bilayer. A methodology is developed whereby NEMD can be effectively used to calculate material properties for complex systems that undergo long time-scale conformational changes. It is found that the bulk modulus upon expansion from a zero stress state agrees well with experimental estimates. However, it is also found that the modulus upon contraction from a zero stress state is larger. From a molecular perspective, it is possible to explain this phenomena by examining the molecular origins of the pressure response. The finding that the two moduli are not equal upon compression and expansion is in apparent contradiction to osmotic stress experiments where the area modulus was found to be the same upon expansion and contraction. This issue is addressed. PMID- 11867441 TI - The brownian ratchet and power stroke models for posttranslational protein translocation into the endoplasmic reticulum. AB - A quantitative analysis of experimental data for posttranslational translocation into the endoplasmic reticulum is performed. This analysis reveals that translocation involves a single rate-limiting step, which is postulated to be the release of the signal sequence from the translocation channel. Next, the Brownian ratchet and power stroke models of translocation are compared against the data. The data sets are simultaneously fit using a least-squares criterion, and both models are found to accurately reproduce the experimental results. A likelihood ratio test reveals that the optimal fit of the Brownian ratchet model, which contains one fewer free parameter, does not differ significantly from that of the power stroke model. Therefore, the data considered here cannot be used to reject this import mechanism. The models are further analyzed using the estimated parameters to make experimentally testable predictions. PMID- 11867442 TI - Piezoelectric reciprocal relationship of the membrane motor in the cochlear outer hair cell. AB - It has been shown that the membrane motor in the outer hair cell is driven by the membrane potential. Here we examine whether the motility satisfies the reciprocal relationship, the characteristic of piezoelectricity, by measuring charge displacement induced by stretching the cell with known force. The efficiency of inducing charge displacement was membrane potential dependent. The maximum efficiency of inducing charge displacement by force was approximately 20 fC/nN for 50-microm-long lateral membrane. The efficiency per cell stretching was 0.1 pC/microm. We found that these values are consistent with the reciprocal relationship based on the voltage sensitivity of approximately 20 nm/mV for 50 microm-long cell and force production of 0.1 nN/mV by the cell. We can thus conclude that the membrane motor in the outer hair cell satisfies a necessary condition for piezoelectricity and that the hair cell's piezoelectric coefficient of 20 fC/nN is four orders of magnitude greater than the best man-made material. PMID- 11867443 TI - Model of magnetic field-induced mitotic apparatus reorientation in frog eggs. AB - Recent experiments have shown that intense static magnetic fields can alter the geometry of the early cell cleavages of Xenopus laevis eggs. The changes depend on field orientation, strength, and timing. We present a model that qualitatively accounts for these effects and which presumes that the structures involved in cell division are cylindrically symmetric and diamagnetically anisotropic and that the geometry of the centrosome replication and spreading processes dictates the nominal cleavage geometry. Within this model, the altered cleavage geometry results from the magnetic field-induced realignment of mitotic structures, which causes a realignment of the centrosome replication and spreading processes. PMID- 11867444 TI - Effects of quercetin on single Ca(2+) release channel behavior of skeletal muscle. AB - Quercetin, a bioflavonoid, is known to affect Ca(2+) fluxes in sarcoplasmic reticulum, although its direct effect on Ca(2+) release channel (CRC) in sarcoplasmic reticulum has remained to be elucidated. The present study examined the effect of quercetin on the behavior of single skeletal CRC in planar lipid bilayer. The effect of caffeine was also studied for comparison. At very low [Ca(2+)](cis) (80 pM), quercetin activated CRC marginally, whereas at elevated [Ca(2+)](cis) (10 microM), both open probability (P(o)) and sensitivity to the drug increased markedly. Caffeine showed a similar tendency. Analysis of lifetimes for single CRC showed that quercetin and caffeine led to different mean open-time and closed-time constants and their proportions. Addition of 10 microM ryanodine to CRC activated by quercetin or caffeine led to the typical subconductance state (approximately 54%) and a subsequent addition of 5 microM ruthenium red completely blocked CRC activity. When 6 microM quercetin and 3 mM caffeine were added together to the cis side of CRC, a time-dependent increase of P(o) was observed (from mode 1 (0.376 +/- 0.043, n = 5) to mode 2 (0.854 +/- 0.062, n = 5)). On the other hand, no further activation was observed when quercetin was added after caffeine. Quercetin affected only the ascending phase of the bell-shaped Ca(2+) activation/inactivation curve, whereas caffeine affected both ascending and descending phases. [(3)H]ryanodine binding to sarcoplasmic reticulum showed that channel activity increased more by both quercetin and caffeine than by caffeine alone. These characteristic differences in the modes of activation of CRC by quercetin and caffeine suggest that the channel activation mechanisms and presumably the binding sites on CRC are different for the two drugs. PMID- 11867445 TI - Cysteine residues in the nucleotide binding domains regulate the conductance state of CFTR channels. AB - Gating of cystic fibrosis transmembrane conductance regulator (CFTR) channels requires intermolecular or interdomain interactions, but the exact nature and physiological significance of those interactions remains uncertain. Subconductance states of the channel may result from alterations in interactions among domains, and studying mutant channels enriched for a single conductance type may elucidate those interactions. Analysis of CFTR channels in inside-out patches revealed that mutation of cysteine residues in NBD1 and NBD2 affects the frequency of channel opening to the full-size versus a 3-pS subconductance. Mutating cysteines in NBD1 resulted in channels that open almost exclusively to the 3-pS subconductance, while mutations of cysteines in NBD2 decreased the frequency of subconductance openings. Wild-type channels open to both size conductances and make fast transitions between them within a single open burst. Full-size and subconductance openings of both mutant and wild-type channels are similarly activated by ATP and phosphorylation. However, the different size conductances open very differently in the presence of a nonhydrolyzable ATP analog, with subconductance openings significantly shortened by ATPgammaS, while full-size channels are locked open. In wild-type channels, reducing conditions increase the frequency and decrease the open time of subconductance channels, while oxidizing conditions decrease the frequency of subconductance openings. In contrast, in the cysteine mutants studied, altering redox potential has little effect on gating of the subconductance. PMID- 11867446 TI - Outer and central charged residues in DIVS4 of skeletal muscle sodium channels have differing roles in deactivation. AB - We tested the effects of charge-neutralizing mutations of the eight arginine residues in DIVS4 of the rat skeletal muscle sodium channel (rNa(V)1.4) on deactivation gating from the open and inactivated states. We hypothesized that neutralization of outer or central charges would accelerate the I-to-C transition as measured by recovery delay because these represent a portion of the immobilizable charge. R1Q abbreviated recovery delay as a consequence of reduced charge content. R4Q increased delay, whereas R5Q abbreviated delay, and charge substitutions at these residues indicated that each effect was allosteric. We also hypothesized that neutralization of any residue in DIVS4 would slow the O-to C transition with reduction in positive charge. Reduction in charge at R1, and to a lesser extent at R5, slowed open-state deactivation, while charge neutralizations at R2, R3, R4, R6, and R7 accelerated open-state deactivation. Our findings suggest that arginine residues in DIVS4 in rNa(V)1.4 have differing roles in channel closure from open and inactivated states. Furthermore, they suggest that deactivation in DIVS4 is regulated by charge interaction between the electrical field with the outermost residue, and by local allosteric interactions imparted by central charges. PMID- 11867449 TI - On the origin of closing flickers in gramicidin channels: a new hypothesis. AB - The submillisecond closing events (flickers) and the single channel conductances to protons (g(H)) were studied in native gramicidin A (gA) and in the SS and RR diastereoisomers of dioxolane-linked gA channels in planar bilayers. Bilayers were formed from glycerylmonooleate (GMO) in various solvents. In GMO/decane (thick) bilayers, the largest flicker frequency occurred in the SS channel (39 s( 1)), followed by the RR (4 s(-1)) and native gA channels (3 s(-1)). These frequencies were attenuated in GMO/squalene (thin) bilayers by 100-, 30-, and 70 fold in the SS, RR, and native gA channels, respectively. In thin bilayers, the average burst duration of native gA channels was 30-fold longer than in thick bilayers. The RR dioxolane-linked gA dimer "inactivated" in GMO/decane but not in squalene-containing bilayers. The mean closed time of flickers (approximately 0.12 ms) was essentially the same in various gA channels. In thin bilayers, g(H) values were larger by approximately 10% (SS), 30% (RR), and 20% (native gA) in relation to thick bilayers. It is concluded that flickers are not related to pre dissociation or dissociation states of gA monomers, and do not seem to be caused by intrinsic conformational changes of channel proteins. It is proposed that flickers are caused by undulations of the bilayer that obliterate the openings of gA channels. Differences between flicker frequencies in various gA channels are likely to result from variations in channel geometries at the bilayer/channel interface. The smaller g(H) in thick bilayers suggests that the deformation of these bilayers around the gA channel creates a diffusional pathway next to the mouths of the channel that is longer and more restrictive than in thin GMO bilayers. A possible molecular interpretation for these effects is attempted. PMID- 11867447 TI - Polyanions decelerate the kinetics of positively charged gramicidin channels as shown by sensitized photoinactivation. AB - The effects of different anionic polymers on the kinetic properties of ionic channels formed by neutral gramicidin A (gA) and its positively charged analogs gramicidin-tris(2-aminoethyl)amine (gram-TAEA) and gramicidin-ethylenediamine (gram-EDA) in a bilayer lipid membrane were studied using a method of sensitized photoinactivation. The addition of Konig's polyanion caused substantial deceleration of the photoinactivation kinetics of gram-TAEA channels, which expose three positive charges to the aqueous phase at both sides of the membrane. In contrast, channels formed of gram-EDA, which exposes one positive charge, and neutral gA channels were insensitive to Konig's polyanion. The effect strongly depended on the nature of the polyanion added, namely: DNA, RNA, polyacrylic acid, and polyglutamic acid were inactive, whereas modified polyacrylic acid induced deceleration of the channel kinetics at high concentrations. In addition, DNA was able to prevent the action of Konig's polyanion. In single-channel experiments, the addition of Konig's polyanion resulted in the appearance of long lived gram-TAEA channels. The deceleration of the gram-TAEA channel kinetics was ascribed to electrostatic interaction of the polyanion with gram-TAEA that reduces the mobility of gram-TAEA monomers and dimers in the membrane via clustering of channels. PMID- 11867450 TI - Sustained overexpression of IGF-1 prevents age-dependent decrease in charge movement and intracellular Ca(2+) in mouse skeletal muscle. AB - In this work we tested the hypothesis that transgenic sustained overexpression of IGF-1 prevents age-dependent decreases in charge movement and intracellular Ca(2+) in skeletal muscle fibers. To this end, short flexor digitorum brevis (FDB) muscle fibers from 5-7- and 21-24-month-old FVB (wild-type) and S1S2 (IGF-1 transgenic) mice were studied. Fibers were voltage-clamped in the whole-cell configuration of the patch-clamp technique according to described procedures (Wang, Z. M., M. L. Messi, and O. Delbono. 1999. Biophys. J. 77:2709-2716). Charge movement and intracellular Ca(2+) concentration were recorded simultaneously. The maximum charge movement (Q(max)) recorded in young wild-type and transgenic mice was (mean +/- SEM, in nC microF(-1)): 52 +/- 2.1 (n = 46) and 54 +/- 1.9 (n = 38) (non-significant, ns), respectively, whereas in old wild-type and old transgenic mice the values were 36 +/- 2.1 (n = 32) and 49 +/- 2.3 (n = 35), respectively (p < 0.01). The peak intracellular calcium [Ca(2+)](i) recorded in young wild-type and transgenic mice was (in muM): 14.5 +/- 0.9 and 16 +/- 2.1 (ns), whereas in old wild-type and transgenic mice the values were 9.9 +/- 0.1 and 14 +/- 1.1 (p < 0.01), respectively. No significant changes in the voltage distribution or steepness of the Q-V or [Ca(2+)]-V relationship were found. These data support the concept that overexpression of IGF-1 in skeletal muscle prevents age-dependent reduction in charge movement and peak [Ca(2+)](i). PMID- 11867448 TI - Imperatoxin a enhances Ca(2+) release in developing skeletal muscle containing ryanodine receptor type 3. AB - Most adult mammalian skeletal muscles contain only one isoform of ryanodine receptor (RyR1), whereas neonatal muscles contain two isoforms (RyR1 and RyR3). Membrane depolarization fails to evoke calcium release in muscle cells lacking RyR1, demonstrating an essential role for this isoform in excitation-contraction coupling. In contrast, the role of RyR3 is unknown. We studied the participation of RyR3 in calcium release in wild type (containing both RyR1 and RyR3 isoforms) and RyR3-/- (containing only RyR1) myotubes in the presence or absence of imperatoxin A (IpTxa), a high-affinity agonist of ryanodine receptors. IpTxa significantly increased the amplitude and the rate of release only in wild-type myotubes. Calcium currents, recorded simultaneously with the transients, were not altered with IpTxa treatment. [(3)H]ryanodine binding to RyR1 or RyR3 was significantly increased in the presence of IpTxa. Additionally, IpTxa modified the gating and conductance level of single RyR1 or RyR3 channels when studied in lipid bilayers. Our data show that IpTxa can interact with both RyRs and that RyR3 is functional in myotubes and it can amplify the calcium release signal initiated by RyR1, perhaps through a calcium-induced mechanism. In addition, our data indicate that when RyR3-/- myotubes are voltage-clamped, the effect of IpTxa is not detected because RyR1s are under the control of the dihydropyridine receptor. PMID- 11867452 TI - Lateral diffusion coefficients in membranes measured by resonance energy transfer and a new algorithm for diffusion in two dimensions. AB - We describe measurements of lateral diffusion in membranes using resonance energy transfer. The donor was a rhenium (Re) metal-ligand complex lipid, which displays a donor decay time near 3 micros. The long donor lifetime resulted in an ability to measure lateral diffusion coefficient below 10(-8) cm(2)/s. The donor decay data were analyzed using a new numerical algorithm for calculation of resonance energy transfer for donors and acceptors randomly distributed in two dimensions. An analytical solution to the diffusion equation in two dimensions is not known, so the equation was solved by the relaxation method in Laplace space. This algorithm allows the donor decay in the absence of energy transfer to be multiexponential. The simulations show that mutual lateral diffusion coefficients of the donor and acceptor on the order of 10(-8) cm(2)/s are readily recovered from the frequency-domain data with donor decay times on the microsecond timescale. Importantly, the lateral diffusion coefficients and acceptor concentrations can be recovered independently despite correlation between these parameters. This algorithm was tested and verified using the donor decays of a long lifetime rhenium lipid donor and a Texas red-lipid acceptor. Lateral diffusion coefficients ranged from 4.4 x 10(-9) cm(2)/s in 1,2-dimyristoyl-sn glycero-3-[phospho-rac-(1-glycerol)] (DMPG) at 10 degrees C to 1.7 x 10(-7) cm(2)/s in 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) at 35 degrees C. These results demonstrated the possibility of direct measurements of lateral diffusion coefficients using microsecond decay time luminophores. PMID- 11867451 TI - An improved method for real-time monitoring of membrane capacitance in Xenopus laevis oocytes. AB - Measurements of membrane capacitance (C(m)) in Xenopus laevis oocytes offer unique experimental possibilities but are difficult to perform with current methods. To improve C(m) measurements in the two-electrode voltage clamp (TEVC) mode, we developed a paired-ramp protocol and tested its performance in a model circuit (with tunable C(m), membrane resistance R(m), and series resistance R(s)) and in Xenopus oocytes. In the cell model and with R(s) = 0 Omega, inaccuracy of C(m) estimates was <1% under widely varying conditions (R(m) ranging from 100 to 2000 kOmega, and C(m) from 50 to 1000 nF). With R(s) > 0 Omega, C(m) was underestimated by a relative error epsilon closely approximated as epsilon approximate 2 x R(s)/(R(s) + R(m)), in keeping with the theoretical prediction. Thus, epsilon may be neglected under standard conditions or, under extreme conditions, corrected for if R(s) is known. Relative imprecision of C(m) estimates was small, independent of R(s), and inversely related to C(m) (<1.5% at 50 nF, <0.4% at 200 nF). Averaging allowed reliable detection of C(m) deviations from 200 nF of 0.1 nF, i.e., 0.05%. In Xenopus oocytes, we could resolve C(m) changes that were small (e.g., DeltaC(m) approximate 2 nF upon 100 muM 8-Br cAMP), fast (e.g., DeltaC(m)/Deltat approximate 20nF/30s upon 1 muM phorbol myristate acetate (PMA)) or extended and complex (e.g., fast increase, followed by prolonged C(m) decrease upon 1 muM PMA). Rapidly alternating between paired ramps and a second, step protocol allowed quasi-simultaneous monitoring of additional electrical parameters such as R(m), slope conductance g(m), and reversal potential E(rev). Taken together, our method is suited to monitor C(m) in Xenopus oocytes conveniently, with high temporal resolution, accuracy and precision, and in parallel with other electrical parameters. Thus, it may be useful for the study of endo- and exocytosis and of membrane protein regulation and for electrophysiological high-throughput screening. PMID- 11867453 TI - Normal and lateral forces between lipid covered solids in solution: correlation with layer packing and structure. AB - We report on the normal and lateral forces between controlled-density mono- and bilayers of phospholipid co-adsorbed onto hydrophobic and hydrophilic solid supports, respectively. Interactions between 1,2-dioleoyl-sn-glycero-3 phosphocholine layers were measured using an atomic force microscope. Notable features of the normal force curves (barrier heights and widths) were found to correlate with the thickness and density of the supported lipid layers. The friction and normal force curves were also found interrelated. Thus, very low friction values were measured as long as the supported layer(s) resisted the normal pressure of the tip. However, as the applied load exceeded the critical value needed for puncturing the layers, the friction jumped to values close to those recorded between bare surfaces. The lipid layers were self-healing between measurements, but a significant hysteresis was observed in the force curves measured on approach and retraction, respectively. The study shows the potential of using atomic force microscopy for lipid layer characterization both with respect to structure and interactions. It further shows the strong lubricating effect of adsorbed lipid layers and how this varies with surface density of lipids. The findings may have important implications for the issue of joint lubrication. PMID- 11867455 TI - Nuclear Overhauser enhancement spectroscopy cross-relaxation rates and ethanol distribution across membranes. AB - Measurement of nuclear Overhauser enhancement spectroscopy cross-relaxation rates between ethanol and palmitoyloleoylphosphatidylcholine bilayers was combined with atomic-level molecular dynamics simulations. The molecular dynamics trajectories yielded autocorrelation functions of proton dipole-dipole interactions, and, consequently, relaxation times and cross-relaxation rates. These analyses allow the measured cross-relaxation rates to be interpreted in terms of relative interaction strengths with the various segments of the lipid molecule. We determined that cross-relaxation between ethanol and specific lipid resonances is primarily determined by the sites of interaction with some modulation due to lipid disorder and to local differences in intramolecular lipid dynamics. The rates scale linearly with the lifetime of temporary ethanol-lipid associations. Ethanol interacts with palmitoyloleoylphosphatidylcholine bilayers primarily via hydrophilic interactions, in particular the formation of hydrogen bonds to the lipid phosphate group. There is a weak contribution to binding from hydrophobic interaction with lipid chain segments near the glycerol. However, the strength of hydrophobic interactions is insufficient to compensate for the energetic loss of locating ethanol in an exclusively hydrophobic environment, resulting in a probability of locating ethanol in the bilayer center that is three orders of magnitude lower than locating ethanol at the lipid/water interface. The low cross relaxation rates between terminal methyl protons of hydrocarbon chains and ethanol are as much the result of infrequent chain upturns as of brief excursions of ethanol into the region of lipid hydrocarbon chains near the glycerol. The combination of nuclear magnetic resonance measurements and molecular dynamics simulations offers a general pathway to study the interaction of small molecules with the lipid matrix at atomic resolution. PMID- 11867454 TI - Membrane tether formation from outer hair cells with optical tweezers. AB - Optical tweezers were used to characterize the mechanical properties of the outer hair cell (OHC) plasma membrane by pulling tethers with 4.5-microm polystyrene beads. Tether formation force and tether force were measured in static and dynamic conditions. A greater force was required for tether formations from OHC lateral wall (499 +/- 152 pN) than from OHC basal end (142 +/- 49 pN). The difference in the force required to pull tethers is consistent with an extensive cytoskeletal framework associated with the lateral wall known as the cortical lattice. The apparent plasma membrane stiffness, estimated under the static conditions by measuring tether force at different tether length, was 3.71 pN/microm for OHC lateral wall and 4.57 pN/microm for OHC basal end. The effective membrane viscosity was measured by pulling tethers at different rates while continuously recording the tether force, and estimated in the range of 2.39 to 5.25 pN x s/microm. The viscous force most likely results from the viscous interactions between plasma membrane lipids and the OHC cortical lattice and/or integral membrane proteins. The information these studies provide on the mechanical properties of the OHC lateral wall is important for understanding the mechanism of OHC electromotility. PMID- 11867456 TI - Characterization of the thermotropic behavior and lateral organization of lipid peptide mixtures by a combined experimental and theoretical approach: effects of hydrophobic mismatch and role of flanking residues. AB - A combined experimental and theoretical study was performed on a series of mixtures of dipalmitoylphosphatidylcholine (DPPC) and synthetic peptides to investigate their thermotropic behavior and lateral organization. The experimental study was based on differential scanning calorimetry (DSC) and phosphorous nuclear magnetic resonance ((31)P-NMR) techniques; the theoretical study was based on calculations on a microscopic molecular interaction model, where the lipid-peptide interaction is built on the hydrophobic matching principle. The chosen peptides, WALP and KALP, consist of a hydrophobic stretch, of variable length, of alternating leucine and alanine residues, flanked on both ends with tryptophan and lysine residues, respectively. By systematically varying the peptide hydrophobic length it was thus possible to explore different matching conditions between the peptide's hydrophobic length and the lipid bilayer hydrophobic thickness, and to investigate the potential role of flanking residues. The results show that both the WALP and the KALP peptides tend to favor the liquid-crystalline (or fluid) phase of the system; i.e., they tend to depress the main-transition temperature, T(m), of pure DPPC. However, the detailed effects of both peptides on the lateral phase behavior of the lipid-peptide system are dependent on the peptide length and the type of flanking residues. The results suggest that below T(m), the shortest among the WALP and KALP peptides induce gel-fluid phase separation in the system within an extensive temperature composition region. The longer the hydrophobic length of the peptides is, the more narrow this region appears to become. PMID- 11867458 TI - From lanosterol to cholesterol: structural evolution and differential effects on lipid bilayers. AB - Cholesterol is an important molecular component of the plasma membranes of mammalian cells. Its precursor in the sterol biosynthetic pathway, lanosterol, has been argued by Konrad Bloch (Bloch, K. 1965. Science. 150:19-28; 1983. CRC Crit. Rev. Biochem. 14:47-92; 1994. Blonds in Venetian Paintings, the Nine-Banded Armadillo, and Other Essays in Biochemistry. Yale University Press, New Haven, CT.) to also be a precursor in the molecular evolution of cholesterol. We present a comparative study of the effects of cholesterol and lanosterol on molecular conformational order and phase equilibria of lipid-bilayer membranes. By using deuterium NMR spectroscopy on multilamellar lipid-sterol systems in combination with Monte Carlo simulations of microscopic models of lipid-sterol interactions, we demonstrate that the evolution in the molecular chemistry from lanosterol to cholesterol is manifested in the model lipid-sterol membranes by an increase in the ability of the sterols to promote and stabilize a particular membrane phase, the liquid-ordered phase, and to induce collective order in the acyl-chain conformations of lipid molecules. We also discuss the biological relevance of our results, in particular in the context of membrane domains and rafts. PMID- 11867457 TI - Rapid transbilayer movement of spin-labeled steroids in human erythrocytes and in liposomes. AB - The transbilayer movement and distribution of spin-labeled analogs of the steroids androstane (SLA) and cholestane (SLC) were investigated in the human erythrocyte and in liposomes. Membranes were labeled with SLA or SLC, and the analogs in the outer leaflet were selectively reduced at 4C using 6-O phenylascorbic acid. As shown previously, 6-O-phenylascorbic acid reduces rapidly nitroxides exposed on the outer leaflet, but its permeation of membranes is comparatively slow and thus does not interfere with the assay. From the reduction kinetics, we infer that transbilayer movement of SLA in erythrocytes is rapid at 4C with a half-time of approximately 4.3 min and that the probe distributes almost symmetrically between both halves of the plasma membrane. We have no indication that a protein-mediated transport is involved in the rapid transbilayer movement of SLA because 1) pretreatment of erythrocytes with N-ethyl maleimide affected neither flip-flop nor transbilayer distribution of SLA and 2) flip-flop of SLA was also rapid in pure lipid membranes. The transbilayer dynamics of SLC in erythrocyte membranes could not be resolved by our assay. Thus, the rate of SLC flip-flop must be on the order of, or even faster than, that of probe reduction rate on the exoplasmic leaflet (half-time approximately 0.5 min). The results are discussed with regard to the transbilayer dynamics of cholesterol. PMID- 11867459 TI - Application of pressure perturbation calorimetry to lipid bilayers. AB - Pressure perturbation calorimetry (PPC) is a new method that measures the heat consumed or released by a sample after a sudden pressure jump. The heat change can be used to derive the thermal volume expansion coefficient, alpha(V), as a function of temperature and, in the case of phase transitions, the volume change, DeltaV, occurring at the phase transition. Here we present the first report on the application of PPC to determine these quantities for lipid bilayers. We measure the volume changes of the pretransition and main transition of 1,2 dimyristoyl-sn-glycero-3-phosphocholine (DMPC), and the thermal expansivity of the fluid phase of DMPC and of two unsaturated lipids, 1-palmitoyl-2-oleoyl-sn glycero-3-phosphocholine and 1,2-dioleoyl-sn-glycero-3-phosphocholine. The high sensitivity of PPC instrumentation gives accurate data for alpha(V) and DeltaV even upon the application of relatively low pressures of approximately 5 bar. PMID- 11867460 TI - Direct observation of poloxamer 188 insertion into lipid monolayers. AB - P188, a triblock copolymer of the form poly(ethylene oxide)-poly(propylene oxide) poly(ethylene oxide) helps seal electroporated cell membranes, arresting the leakage of intracellular materials from the damaged cells. To explore the nature of the interaction between P188 and cell membranes, we have constructed a model system that assesses the ability of P188 to insert into lipid monolayers. Using concurrent Langmuir isotherm and fluorescence microscopy measurements, we find that P188 changes the phase behavior and morphology of the monolayers. P188 inserts into both dipalmitoylphosphatidlycholine and dipalmitoylphosphatidylglycerol monolayers at surface pressures equal to and lower than approximately 22 mN/m at 30 degrees C; this pressure corresponds to the maximal surface pressure attained by P188 on a pure water subphase. Similar results for the two phospholipids indicate that P188 insertion is not influenced by headgroup electrostatics. Because the equivalent surface pressure of a normal bilayer is on the order of 30 mN/m, the lack of P188 insertion above 22 mN/m further suggests the poloxamer selectively adsorbs into damaged portions of electroporated membranes, thereby localizing its effect. P188 is also found to be "squeezed out" of the monolayers at high surface pressures, suggesting a mechanism for the cell to be rid of the poloxamer when the membrane is restored. PMID- 11867461 TI - Molecular dynamics simulation of proton transport near the surface of a phospholipid membrane. AB - The structural and dynamical properties of a hydrated proton near the surface of DMPC membrane were studied using a molecular dynamics simulation. The proton transport between water molecules was modeled using the second generation multistate empirical valence bond model. The proton diffusion was found to be inhibited at the membrane surface. The potential of mean force for the proton adsorption to the membrane surface and its release back into the bulk water was also determined, yielding a small barrier in each direction. An efficient algorithm for Ewald summation calculations for the multistate empirical valence bond model is also introduced. PMID- 11867463 TI - Role of the Na(+)-Ca(2+) exchanger as an alternative trigger of CICR in mammalian cardiac myocytes. AB - Ca(2+) influx through the L-type Ca(2+) channels is the primary pathway for triggering the Ca(2+) release from the sarcoplasmic reticulum (SR). However, several observations have shown that Ca(2+) influx via the reverse mode of the Na(+)-Ca(2+) exchanger current (I(Na-Ca)) could also trigger the Ca(2+) release. The aim of the present study was to quantitate the role of this alternative pathway of Ca(2+) influx using a mathematical model. In our model 20% of the fast sodium channels and the Na(+)-Ca(2+) exchanger molecules are located in the restricted subspace between the sarcolemma and the SR where triggering of the calcium-induced calcium release (CICR) takes place. After determining the strengths of the alternative triggers with simulated voltage-clamps in varied membrane voltages and resting [Na](i) values, we studied the CICR in simulated action potentials, where fast sodium channel current contributes [Na](i) of the subspace. In low initial [Na](i) the Ca(2+) influx via the L-type Ca(2+) channels is the major trigger for Ca(2+) release from the SR, and the Ca(2+) influx via the reverse mode of the Na(+)-Ca(2+) exchanger cannot trigger the CICR. However, depending on the initial [Na](i), the contribution of the Ca(2+) entry via the exchanger may account for 25% (at [Na](i) = 10 mM) to nearly 100% ([Na](i) = 30 mM) of the trigger Ca(2+). The shift of the main trigger from L-type calcium channels to the exchanger reduced the delay between the action potential upstroke and the intracellular calcium transient. This may contribute to the function of the myocyte in physiological situations where [Na](i) is elevated. These main results remain the same when using different estimates for the most crucial parameters in the modeling or different models for the exchanger. PMID- 11867462 TI - Structure, composition, and peptide binding properties of detergent soluble bilayers and detergent resistant rafts. AB - Lipid bilayers composed of unsaturated phosphatidylcholine (PC), sphingomyelin (SM), and cholesterol are thought to contain microdomains that have similar detergent insolubility characteristics as rafts isolated from cell plasma membranes. We chemically characterized the fractions corresponding to detergent soluble membranes (DSMs) and detergent resistant membranes (DRMs) from 1:1:1 PC:SM:cholesterol, compared the binding properties of selected peptides to bilayers with the compositions of DSMs and DRMs, used differential scanning calorimetry to identify phase transitions, and determined the structure of DRMs with x-ray diffraction. Compared with the equimolar starting material, DRMs were enriched in both SM and cholesterol. Both transmembrane and interfacial peptides bound to a greater extent to DSM bilayers than to DRM bilayers, likely because of differences in the mechanical properties of the two bilayers. Thermograms from 1:1:1 PC:SM:cholesterol from 3 to 70 degrees C showed no evidence for a liquid ordered to liquid-disordered phase transition. Over a wide range of osmotic stresses, each x-ray pattern from equimolar PC:SM:cholesterol or DRMs contained a broad wide-angle band at 4.5 A, indicating that the bilayers were in a liquid crystalline phase, and several sharp low-angle reflections that indexed as orders of a single lamellar repeat period. Electron density profiles showed that the total bilayer thickness was 57 A for DRMs, which was approximately 5 A greater than that of 1:1:1 PC:SM:cholesterol and 10 A greater than the thickness of bilayers with the composition of DSMs. These x-ray data provide accurate values for the widths of raft and nonraft bilayers that should be important in understanding mechanisms of protein sorting by rafts. PMID- 11867464 TI - Mammalian cardiac muscle thick filaments: their periodicity and interactions with actin. AB - Cardiac muscle has been extensively studied, but little information is available on the detailed macromolecular structure of its thick filament. To elucidate the structure of these filaments I have developed a procedure to isolate the cardiac thick filaments for study by electron microscopy and computer image analysis. This procedure uses chemical skinning with Triton X-100 to avoid contraction of the muscle that occurs using the procedures previously developed for isolation of skeletal muscle thick filaments. The negatively stained isolated filaments appear highly periodic, with a helical repeat every third cross-bridge level (43 nm). Computed Fourier transforms of the filaments show a strong set of layer lines corresponding to a 43-nm near-helical repeat out to the 6th layer line. Additional meridional reflections extend to at least the 12th layer line in averaged transforms of the filaments. The highly periodic structure of the filaments clearly suggests that the weakness of the layer lines in x-ray diffraction patterns of heart muscle is not due to an inherently more disordered cross-bridge arrangement. In addition, the isolated thick filaments are unusual in their strong tendency to remain bound to actin by anti-rigor oriented cross bridges (state II or state III cross-bridges) under relaxing conditions. PMID- 11867465 TI - Sustained release of calcium elicited by membrane depolarization in ryanodine injected mouse skeletal muscle fibers. AB - The effect of micromolar intracellular levels of ryanodine was tested on the myoplasmic free calcium concentration ([Ca(2+)](i)) measured from a portion of isolated mouse skeletal muscle fibers voltage-clamped at -80 mV. When ryanodine injected fibers were transiently depolarized to 0 mV, the early decay phase of [Ca(2+)](i) upon membrane repolarization was followed by a steady elevated [Ca(2+)](i) level. This effect could be qualitatively well simulated, assuming that ryanodine binds to release channels that open during depolarization and that ryanodine-bound channels do not close upon repolarization. The amplitude of the postpulse [Ca(2+)](i) elevation depended on the duration of the depolarization, being hardly detectable for pulses shorter than 100 ms, and very prominent for duration pulses of seconds. Within a series of consecutive pulses of the same duration, the effect of ryanodine produced a staircase increase in resting [Ca(2+)](i), the slope of which was approximately twice larger for depolarizations to 0 or +10 mV than to -30 or -20 mV. Overall results are consistent with the "open-locked" state because of ryanodine binding to calcium release channels that open during depolarization. Within the voltage-sensitive range of calcium release, increasing either the amplitude or the duration of the depolarization seems to enhance the fraction of release channels accessible to ryanodine. PMID- 11867466 TI - Ca(2+)-induced movement of tropomyosin in skeletal muscle thin filaments observed by multi-site FRET. AB - To obtain information on Ca(2+)-induced tropomyosin (Tm) movement in Ca(2+) regulated muscle thin filaments, frequency-domain fluorescence energy transfer data were collected between 5-(2-iodoacetyl-amino-ethyl-amino)naphthalene-1 sulfonic acid at Cys-190 of Tm and phalloidin-tetramethylrhodamine B isothiocyanate bound to F-actin. Two models were used to fit the experimental data: an atomic coordinate (AC) model coupled with a search algorithm that varies the position and orientation of Tm on F-actin, and a double Gaussian distance distribution (DD) model. The AC model showed that little or no change in transfer efficiency is to be expected between different sites on F-actin and Tm if Ca(2+) causes azimuthal movement of Tm of the magnitude suggested by structural data (C. Xu, R. Craig, L. Tobacman, R. Horowitz, and W. Lehman. 1999. Biophys. J. 77:985 992). However, Ca(2+) produced a small but significant change in our phase/modulation versus frequency data, showing that changes in lifetime decay can be detected even when a change of the steady-state transfer efficiency is very small. A change in Tm azimuthal position of 17 on the actin filament obtained with the AC model indicates that solution data are in reasonable agreement with EM image reconstruction data. In addition, the data indicate that Tm also appears to rotate about its axis, resulting in a rolling motion over the F-actin surface. The DD model showed that the distance from one of the two chains of Tm to F-actin was mainly affected, further verifying that Ca(2+) causes Tm to roll over the F-actin surface. The width of the distance distributions indicated that the position of Tm in absence and in presence of Ca(2+) is well defined with appreciable local flexibility. PMID- 11867467 TI - Unzipping DNA with optical tweezers: high sequence sensitivity and force flips. AB - Force measurements are performed on single DNA molecules with an optical trapping interferometer that combines subpiconewton force resolution and millisecond time resolution. A molecular construction is prepared for mechanically unzipping several thousand-basepair DNA sequences in an in vitro configuration. The force signals corresponding to opening and closing the double helix at low velocity are studied experimentally and are compared to calculations assuming thermal equilibrium. We address the effect of the stiffness on the basepair sensitivity and consider fluctuations in the force signal. With respect to earlier work performed with soft microneedles, we obtain a very significant increase in basepair sensitivity: presently, sequence features appearing at a scale of 10 basepairs are observed. When measured with the optical trap the unzipping force exhibits characteristic flips between different values at specific positions that are determined by the base sequence. This behavior is attributed to bistabilities in the position of the opening fork; the force flips directly reflect transitions between different states involved in the time-averaging of the molecular system. PMID- 11867468 TI - Intrinsic conformational energetics associated with the glycosyl torsion in DNA: a quantum mechanical study. AB - The glycosyl torsion (chi) in nucleic acids has long been recognized to be a major determinant of their conformational properties. chi torsional energetics were systematically mapped in deoxyribonucleosides using high-level quantum mechanical methods, for north and south sugar puckers and with gamma in the g(+) and trans conformations. In all cases, the syn conformation is found higher in energy than the anti. When gamma is changed from g(+) to trans, the anti orientation of the base is strongly destabilized, and the energy difference and barrier between anti and syn are significantly decreased. The barrier between anti and syn in deoxyribonucleosides is found to be less than 10 kcal/mol and tends to be lower with purines than with pyrimidines. With gamma = g(+)/chi = anti, a south sugar yields a significantly broader energy well than a north sugar with no energy barrier between chi values typical of A or B DNA. Contrary to the prevailing view, the syn orientation is not more stable with south puckers than with north puckers. The syn conformation is significantly more energetically accessible with guanine than with adenine in 5-nucleotides but not in nucleosides. Analysis of nucleic acid crystal structures shows that gamma = trans/chi = anti is a minor but not negligible conformation. Overall, chi appears to be a very malleable structural parameter with the experimental chi distributions reflecting, to a large extent, the associated intrinsic torsional energetics. PMID- 11867470 TI - Two-photon excited fluorescence from higher electronic states of chlorophylls in photosynthetic antenna complexes: a new approach to detect strong excitonic chlorophyll a/b coupling. AB - Stepwise two-photon excitation of chlorophyll a and b in the higher plant main light-harvesting complex (LHC II) and the minor complex CP29 (as well as in organic solution) with 100-fs pulses in the Q(y) region results in a weak blue fluorescence. The dependence of the spectral shape of the blue fluorescence on excitation wavelength offers a new approach to elucidate the long-standing problem of the origin of spectral "chlorophyll forms" in pigment-protein complexes, in particular the characterization of chlorophyll a/b-heterodimers. As a first result we present evidence for the existence of strong chlorophyll a/b interactions (excitonically coupled transitions at 650 and 680 nm) in LHC II at ambient temperature. In comparison with LHC II, the experiments with CP29 provide further evidence that the lowest energy chlorophyll a transition (at approximately 680 nm) is not excitonically coupled to chlorophyll b. PMID- 11867469 TI - Entrapment and condensation of DNA in neutral reverse micelles. AB - DNA condensation and compaction is induced by a variety of condensing agents such as polycations. The present study analyzed the structure of plasmid DNA (DNA) in the small inner space of reverse micelles formed from nonionic surfactants (isotropic phase). Spectroscopic studies indicated that DNA was dissolved in an organic solvent in the presence of a neutral detergent. Fluorescent quenching of ethidium bromide and of rhodamine covalently attached to DNA suggested that the DNA within neutral, reverse micelles was condensed. Circular dichroism indicated that the DNA structure was C form (member of B family) and not the dehydrated A form. Concordantly, NMR experiments indicated that the reverse micelles contained a pool of free water, even at a ratio of water to surfactant (Wo) of 3.75. Electron microscopic analysis also indicated that the DNA was in a ring-like structure, probably toroids. Atomic force microscopic images also revealed small, compact particles after the condensed DNA structures were preserved using an innovative cross-linking strategy. In the lamellar phase, the DNA was configured in long strands that were 20 nm in diameter. Interestingly, such DNA structures, reminiscent of "nanowires," have apparently not been previously observed. PMID- 11867471 TI - Pathways for energy transfer in the core light-harvesting complexes CP43 and CP47 of photosystem II. AB - The pigment-protein complexes CP43 and CP47 transfer excitation energy from the peripheral antenna of photosystem II toward the photochemical reaction center. We measured the excitation dynamics of the chlorophylls in isolated CP43 and CP47 complexes at 77 K by time-resolved absorbance-difference and fluorescence spectroscopy. The spectral relaxation appeared to occur with rates of 0.2-0.4 ps and 2-3 ps in both complexes, whereas an additional relaxation of 17 ps was observed only in CP47. Using the 3.8-A crystal structure of the photosystem II core complex from Synechococcus elongatus (A. Zouni, H.-T. Witt, J. Kern, P. Fromme, N. Krauss, W. Saenger, and P. Orth, 2001, Nature, 409:739-743), excitation energy transfer kinetics were calculated and a Monte Carlo simulation of the absorption spectra was performed. In both complexes, the rate of 0.2-0.4 ps can be ascribed to excitation energy transfer within a layer of chlorophylls near the stromal side of the membrane, and the slower 2-3-ps process to excitation energy transfer to the calculated lowest excitonic state. We conclude that excitation energy transfer within CP43 and CP47 is fast and does not contribute significantly to the well-known slow trapping of excitation energy in photosystem II. PMID- 11867472 TI - Fourier transform infrared study of the effect of different cations on bacteriorhodopsin protein thermal stability. AB - The effect of divalent ion binding to deionized bacteriorhodopsin (dI-bR) on the thermal transitions of the protein secondary structure have been studied by using temperature-dependent Fourier transform infrared (FT-IR) spectroscopy. The native metal ions in bR, Ca(2+), and Mg(2+), which we studied previously, are compared with Mn(2+), Hg(2+), and a large, synthesized divalent organic cation, ((Et)(3)N)(2)Bu(2+). It was found that in all cases of ion regeneration, there is a pre-melting, reversible conformational transition in which the amide frequency shifts from 1665 to 1652 cm(-1). This always occurs at approximately 80 degrees C, independent of which cation is used for the regeneration. The irreversible thermal transition (melting), monitored by the appearance of the band at 1623 cm( 1), is found to occur at a lower temperature than that for the native bR but higher than that for acid blue bR in all cases. However, the temperature for this transition is dependent on the identity of the cation. Furthermore, it is shown that the mechanism of melting of the organic cation regenerated bR is different than for the metal cations, suggesting a difference in the type of binding to the protein (either to different sites or different binding to the same site). These results are used to propose specific direct binding mechanisms of the ions to the protein of deionized bR. PMID- 11867473 TI - Spectroscopic studies on the interaction of a water soluble porphyrin and two drug carrier proteins. AB - The interaction of meso-tetrakis(p-sulfonatophenyl)porphyrin (TSPP) sodium salt to human serum albumin and beta-lactoglobulin was studied by steady-state and dynamic fluorescence at different pH of aqueous solutions. The formation of TSPP J-aggregates and a noncovalent TSPP-protein complex was monitored by fluorescence titrations, which depend on pH and on the protein nature and concentration. The complex between TSPP and protein displays a heterogeneous equilibrium with large changes in the binding strength versus pH. The large reduction of the effective binding constant from pH 2 to 7 suggests that electrostatic interactions are a major contribution to the binding of TSPP to the aforementioned proteins. TSPP aggregates and TSPP-protein complex exhibit circular dichroism induced by the presence of the protein. Circular dichroism spectra in the ultraviolet region show that the secondary structure of both proteins is not extensively affected by the TSPP presence. Protein-TSPP interaction was also examined by following the intrinsic fluorescence of the tryptophan residues of the proteins. Fluorescence quenching by acrylamide and TSPP itself also point to small changes on the protein tertiary structure and a critical distance R(0) approximately 56 A, between tryptophan and bound porphyrin, was estimated using the long distance Forster-type energy transfer formalism. PMID- 11867474 TI - Rapid measurement of protein osmotic second virial coefficients by self interaction chromatography. AB - Weak protein interactions are often characterized in terms of the osmotic second virial coefficient (B(22)), which has been shown to correlate with protein phase behavior, such as crystallization. Traditional methods for measuring B(22), such as static light scattering, are too expensive in terms of both time and protein to allow extensive exploration of the effects of solution conditions on B(22). In this work we have measured protein interactions using self-interaction chromatography, in which protein is immobilized on chromatographic particles and the retention of the same protein is measured in isocratic elution. The relative retention of the protein reflects the average protein interactions, which we have related to the second virial coefficient via statistical mechanics. We obtain quantitative agreement between virial coefficients measured by self-interaction chromatography and traditional characterization methods for both lysozyme and chymotrypsinogen over a wide range of pH and ionic strengths, yet self interaction chromatography requires at least an order of magnitude less time and protein than other methods. The method thus holds significant promise for the characterization of protein interactions requiring only commonly available laboratory equipment, little specialized expertise, and relatively small investments of both time and protein. PMID- 11867476 TI - Stability diagram and unfolding of a modified cytochrome c: what happens in the transformation regime? AB - We determined the stability diagram of a modified cytochrome c protein in a glycerol water mixture by measuring the first and the second moment of the fluorescence from the chromophore as a function of temperature and pressure. Temperature and pressure were varied between 273 and 363 K and 0.0001 and 1 GPa, respectively. The shift of the fluorescence maximum showed a characteristic sigmoid-like pattern from which information on the microscopic processes during unfolding is obtained: as the transformation regime is entered, the fluorescence shows a significant blue shift. The conclusion is that water molecules get into contact with the chromophore. They lead to strong electrostatic contributions in the solvent shift, which counteract the red shifting dispersion interactions. Assuming that there are just two relevant states that determine the stability diagram, the complete set of thermodynamic parameters can be determined from the data. However, under certain pressure-temperature conditions the fluorescence pattern is more complicated, pointing toward reentrant transitions and, possibly, to consecutive steps in the unfolding process. PMID- 11867475 TI - Transient exposure of hydrophobic surface in the photoactive yellow protein monitored with Nile Red. AB - In this study we have investigated binding of the fluorescent hydrophobicity probe Nile Red to the photoactive yellow protein (PYP), to characterize the exposure and accessibility of hydrophobic surface upon formation of the signaling state of this photoreceptor protein. Binding of Nile Red, reflected by a large blue shift and increase in fluorescence quantum yield of the Nile Red emission, is observed exclusively when PYP resides in its signaling state. N-terminal truncation of the protein allows assignment of the region surrounding the chromophore as the site where Nile Red binds to PYP. We also observed a pH dependence of the affinity of Nile Red for pB, which we propose is caused by pH dependent differences of the structure of the signaling state. From a comparative analysis of the kinetics of Nile Red binding and transient absorption changes in the visible region we can conclude that protonation of the chromophore precedes the exposure of a hydrophobic surface near the chromophore binding site, upon formation of the signaling state. Furthermore, the data presented here favor the view that the signaling state is structurally heterogeneous. PMID- 11867477 TI - DNA-binding interactions and conformational fluctuations of Tc3 transposase DNA binding domain examined with single molecule fluorescence spectroscopy. AB - The fluorescent dye tetramethylrhodamine (TMR) was conjugated to a synthetic peptide containing the sequence-specific DNA binding domain of Tc3 transposase. Steady-state and single molecule fluorescence spectroscopy was used to investigate protein conformational fluctuations and the thermodynamics of binding interactions. Evidence is presented to show that the TMR-Tc3 conjugate exists in at least two conformational states. The most stable conformation is one in which the TMR fluorescence is quenched. Upon binding to DNA, the total fluorescence from TMR-Tc3 increases by three- to fourfold. Single molecule measurements of TMR Tc3 bound to DNA shows that this complex also fluctuates between a fluorescent and quenched form. The fluorescent form of the conjugate is stabilized when bound to DNA, and this accounts for part of the increase in total fluorescence. In addition, the inherent photodynamics of the dye itself is also altered (e.g., fluorescent lifetime or triplet yield) in such a way that the total fluorescence from the conjugate bound to DNA is enhanced relative to the unbound form. PMID- 11867479 TI - Research realpolitik. PMID- 11867478 TI - Imaging the electrostatic potential of transmembrane channels: atomic probe microscopy of OmpF porin. AB - The atomic force microscope (AFM) was used to image native OmpF porin and to detect the electrostatic potential generated by the protein. To this end the OmpF porin trimers from Escherichia coli was reproducibly imaged at a lateral resolution of approximately 0.5 nm and a vertical resolution of approximately 0.1 nm at variable electrolyte concentrations of the buffer solution. At low electrolyte concentrations the charged AFM probe not only contoured structural details of the membrane protein surface but also interacted with local electrostatic potentials. Differences measured between topographs recorded at variable ionic strength allowed mapping of the electrostatic potential of OmpF porin. The potential map acquired by AFM showed qualitative agreement with continuum electrostatic calculations based on the atomic OmpF porin embedded in a lipid bilayer at the same electrolyte concentrations. Numerical simulations of the experimental conditions showed the measurements to be reproduced quantitatively when the AFM probe was included in the calculations. This method opens a novel avenue to determine the electrostatic potential of native protein surfaces at a lateral resolution better than 1 nm and a vertical resolution of approximately 0.1 nm. PMID- 11867480 TI - Boxing and the brain. Revisiting chronic traumatic encephalopathy. PMID- 11867481 TI - Should we treat concussion pharmacologically? The need for evidence based pharmacological treatment for the concussed athlete. PMID- 11867482 TI - Summary and agreement statement of the First International Conference on Concussion in Sport, Vienna 2001. Recommendations for the improvement of safety and health of athletes who may suffer concussive injuries. PMID- 11867483 TI - New criteria for female athlete triad syndrome? As osteoporosis is rare, should osteopenia be among the criteria for defining the female athlete triad syndrome? PMID- 11867484 TI - Intensive training in elite young female athletes. Effects of intensive training on growth and maturation are not established. PMID- 11867485 TI - Banning pregnant netballers--is this the answer? PMID- 11867487 TI - Does rugby headgear prevent concussion? Attitudes of Canadian players and coaches. AB - OBJECTIVES: To examine the attitudes of players and coaches to the use of protective headgear, particularly with respect to the prevention of concussion. METHODS: A questionnaire designed to assess attitudes to headgear was administered to 63 players from four different Canadian teams, each representing a different level of play (high school, university, community club, national). In addition, coaches from all four levels were questioned about team policies and their personal opinions about the use of headgear to prevent concussion. RESULTS: Although the players tended to believe that the headgear could prevent concussion (62%), the coaches were less convinced (33%). Despite the players' belief that headgear offers protection against concussion, only a minority reported wearing headgear (27%) and few (24%) felt that its use should be made mandatory. Common reasons for not wearing headgear were "its use is not mandatory", "it is uncomfortable", and "it costs too much". CONCLUSION: Although most players in the study believe that rugby headgear may prevent concussion, only a minority reported wearing it. Coaches tended to be less convinced than the players that rugby headgear can prevent concussion. PMID- 11867486 TI - Who owns the information? Databases of injuries in professional sport are valuable resources which should not suffer confidentiality restraints. PMID- 11867488 TI - Incidence of injury in amateur rugby league sevens. AB - OBJECTIVES: To investigate the incidence, site, and nature of injuries sustained in amateur rugby league sevens tournaments. METHODS: A total of 168 players competing in three amateur rugby league sevens tournaments were studied. All injuries sustained during matches were recorded. Information recorded included the name of the injured player and the time, cause, anatomical site, and nature of the injury. RESULTS: The incidence of injury was 283.5 per 1000 playing hours. Some 40% (113.4 per 1000 playing hours) of all injuries sustained were to the lower limb (chi(2) = 5.3, df = 1, p<0.05). Contusions were the most common type of injury (113.4 per 1000 hours, 40%, chi(2) = 9.5, df = 4, p<0.05), with most (198.4 per 1000 hours, 70%, chi(2) = 31.5, df =4, p<0.001) occurring in physical collisions and tackles. An increasing injury incidence was observed over the first (99.2 per 1000 hours), second (198.4 per 1000 hours), third (347.2 per 1000 hours), and fourth (694.4 per 1000 hours) matches played during the tournaments (chi(2) = 9.2, df = 3, p<0.05). CONCLUSIONS: The results of this study suggest that amateur rugby league sevens tournaments, which require players to compete repeatedly on the same day, may hasten the onset of fatigue and predispose to injury. PMID- 11867489 TI - The impact of face shield use on concussions in ice hockey: a multivariate analysis. AB - OBJECTIVE: To identify specific risk factors for concussion severity among ice hockey players wearing full face shields compared with half face shields (visors). METHODS: A prospective cohort study was conducted during one varsity hockey season (1997-1998) with 642 male ice hockey players (median age 22 years) from 22 teams participating in the Canadian Inter-University Athletics Union. Half of the teams wore full face shields, and half wore half shields (visors) for every practice and game throughout the season. Team therapists and doctors recorded on structured forms daily injury, participation, and information on face shield use for each athlete. The main outcome measure was any traumatic brain injury requiring assessment or treatment by a team therapist or doctor, categorised by time lost from subsequent participation and compared by type of face shield worn. RESULTS: Players who wore half face shields missed significantly more practices and games per concussion (2.4 times) than players who wore full face shields (4.07 sessions (95% confidence interval (CI) 3.48 to 4.74) v 1.71 sessions (95% CI 1.32 to 2.18) respectively). Significantly more playing time was lost by players wearing half shields during practices and games, and did not depend on whether the athletes were forwards or defence, rookies or veterans, or whether the concussions were new or recurrent. In addition, players who wore half face shields and no mouthguards at the time of concussion missed significantly more playing time (5.57 sessions per concussion; 95% CI 4.40 to 6.95) than players who wore half shields and mouthguards (2.76 sessions per concussion; 95% CI 2.14 to 3.55). Players who wore full face shields and mouthguards at the time of concussion lost no playing time compared with 1.80 sessions lost per concussion (95% CI 1.38 to 2.34) for players wearing full face shields and no mouthguards. CONCLUSIONS: The use of a full face shield compared with half face shield by intercollegiate ice hockey players significantly reduced the playing time lost because of concussion, suggesting that concussion severity may be reduced by the use of a full face shield. PMID- 11867490 TI - Perceptions of retired professional soccer players about the provision of support services before and after retirement. AB - OBJECTIVE: To determine the views of retired players about the provision of support services in English professional soccer before and after retirement and to assess the impact of career ending injury on these views. METHOD: An anonymous self administered questionnaire was distributed to 500 former players registered with the English Professional Footballers' Association. The questions asked about personal details, current medical status, reasons for retirement, perceptions of the provision and quality of support services, and use of prophylactic treatments while injured. RESULTS: Of the 500 questionnaires distributed, 185 (37%) were returned. The Professional Footballers' Association provided significantly (p<0.001) more help and advice to retired players on medical, financial, career, and educational matters than any other organisation. Although respondents showed some satisfaction with the provision of medical support, they were significantly (p<0.001) less satisfied with the provision of sports science (23%) and education/welfare (19%) support. Respondents who had retired through injury, however, were more dissatisfied with the provision of all services. Significantly (p<0.001) more respondents agreed with the view that injuries would reduce income earning potential, contribute to medical problems, and restrict the duration of their playing career. Respondents who had been medically diagnosed with osteoarthritis were significantly more likely, at some time, to have regularly received steroid injections while injured. CONCLUSION: The results presented are consistent with other evidence that the provision of injury prevention and socioeconomic services at professional soccer clubs is inadequate. The soccer industry should therefore develop a long term strategy for managing the needs of players who are forced to retire through injury. PMID- 11867491 TI - Epidemiology of injuries in the Australian Football League, seasons 1997-2000. AB - OBJECTIVE: To describe the epidemiology of injuries in the Australian Football League (AFL) over four seasons. METHODS: An injury was defined as "any physical or medical condition that caused a player to miss a match in the regular season." The rationale for this definition was to eliminate a previously noted tendency of team recorders to interpret injury definitions subjectively. Administrative records of injury payments to players who did not play matches determined the occurrence of an injury. RESULTS: The seasonal incidence of new injuries was 39 per club (of 40 players) per season (of 22 matches). The match injury incidence for AFL games was 25.7 injuries per 1000 player hours. The injury prevalence (percentage of players missing through injury in an average week) was 16%. The recurrence rate of injuries was 17%. The most common and prevalent injury was hamstring strain (six injuries per club per season, resulting in 21 missed matches per club per season), followed in prevalence by anterior cruciate ligament and groin injuries. CONCLUSIONS: The injury definition of this study does not produce incidence rates that are complete for all minor injuries. However, the determination of an injury is made by a single entity in exactly the same manner for all teams, which overcomes a significant methodological flaw present in other multiteam injury surveillance systems. PMID- 11867492 TI - Identifying college athletes at risk for pathogenic eating. AB - OBJECTIVES: To evaluate the effectiveness of a discriminant function that predicts risk of pathogenic eating in comparison with a standard self report measure (EAT) and a clinical interview. In addition, to determine the effectiveness of this discriminant function using a variety of collegiate athletes. METHODS: A total of 319 participants were asked to complete a series of self report measures that assessed dietary practices. In addition, anthropometric measures were obtained, and a random sample of 15% participated in a structured clinical interview. RESULTS: Correlational analyses indicated that the discriminant function categorisation of risk was significantly related to both the clinical interview and EAT (p < or =0.05). The discriminant function was accurate in predicting risk category in this diverse group of athletes, particularly with respect to those at low risk (83.1%) and those at high risk (72.7%). CONCLUSION: This information may be helpful in the development of a simple, accessible tool to identify athletes at risk of engaging in pathogenic eating behaviours. PMID- 11867493 TI - Beneficial effects of air inclusions on the performance of ethylene vinyl acetate (EVA) mouthguard material. AB - OBJECTIVE: To investigate the impact characteristics of an ethylene vinyl acetate (EVA) mouthguard material with regulated air inclusions, which included various air cell volumes and wall thickness between air cells. In particular, the aim was to identify the magnitude and direction of forces within the impacts. METHOD: EVA mouthguard material, 4 mm thick and with and without air inclusions, was impacted with a constant force impact pendulum with an energy of 4.4 J and a velocity of 3 m/s. Transmitted forces through the EVA material were measured using an accelerometer, which also allowed the determination of force direction and magnitude within the impacts. RESULTS: Statistically significant reductions in the transmitted forces were observed with all the air inclusion materials when compared with EVA without air inclusions. Maximum transmitted force through one air inclusion material was reduced by 32%. Force rebound was eliminated in one material, and reduced second force impulses were observed in all the air inclusion materials. CONCLUSION: The regulated air inclusions improved the impact characteristics of the EVA mouthguard material, the material most commonly used in mouthguards world wide. PMID- 11867494 TI - Identifying some determinants of "jet lag" and its symptoms: a study of athletes and other travellers. AB - BACKGROUND: Travelling across multiple time zones disrupts normal circadian rhythms and induces "jet lag". Possible effects of this on training and performance in athletes were concerns before the Sydney Olympic Games. OBJECTIVE: To identify some determinants of jet lag and its symptoms. METHODS: A mixture of athletes, their coaches, and academics attending a conference (n = 85) was studied during their flights from the United Kingdom to Australia (two flights with a one hour stopover in Singapore), and for the first six days in Australia. Subjects differed in age, sex, chronotype, flexibility of sleeping habits, feelings of languor, fitness, time of arrival in Australia, and whether or not they had previous experience of travel to Australia. These variables and whether the body clock adjusted to new local time by phase advance or delay were tested as predictors for jet lag and some of its symptoms by stepwise multiple regression analyses. RESULTS: The amount of sleep in the first flight was significantly greater in those who had left the United Kingdom in the evening than the morning (medians of 5.5 hours and 1.5 hours respectively; p = 0.0002, Mann-Whitney), whereas there was no significant difference on the second flight (2.5 hours v 2.8 hours; p = 0.72). Only the severity of jet lag and assessments of sleep and fatigue were commonly predicted significantly (p<0.05) by regression analysis, and then by only some of the variables. Thus increasing age and a later time of arrival in Australia were associated with less jet lag and fatigue, and previous experience of travel to Australia was associated with an earlier time of getting to sleep. Subjects who had adjusted by phase advance suffered worse jet lag during the 5th and 6th days in Australia. CONCLUSIONS: These results indicate the importance of an appropriate choice of itinerary and lifestyle for reducing the negative effects of jet lag in athletes and others who wish to perform optimally in the new time zone. PMID- 11867495 TI - Views of chartered physiotherapists on the psychological content of their practice: a preliminary study in the United Kingdom. AB - BACKGROUND: Although research into the psychological aspects of sports injury is increasing and psychological interventions have been identified as important in the rehabilitation process, few studies have focused on how sports medicine practitioners deal with psychological problems. OBJECTIVE: To investigate the perceptions of English chartered physiotherapists on the psychological content of their practice. METHODS: The Physiotherapist and Sport Psychology Questionnaire (PSPQ) was adapted slightly from the Athletic Trainer and Sport Psychology Questionnaire and used in this study. A survey package comprising a PSPQ, introductory letter, and self addressed envelope was mailed to 179 chartered physiotherapists registered in the England Eastern Region Sports Medicine Directory. RESULTS: In total, 90 (50% response rate) questionnaires were returned. The sample consisted of 67 women and 23 men with a mean (SD) age of 40.1 (5.4) years and 9.2 (3.1) years of experience as chartered physiotherapists. Descriptive statistical and qualitative analysis showed that physiotherapists believed athletes were often psychologically affected by injury. The physiotherapists also reported often using psychological techniques when treating injured athletes, but few reported having access to a sport psychologist for referral. CONCLUSIONS: This research indicates that future physiotherapy education may need more emphasis on the psychological aspect of injury, and seek to increase knowledge on the potential of using psychological interventions within a physiotherapy rehabilitation programme. Furthermore, some form of referral network should be established between chartered physiotherapists and sport psychologists. PMID- 11867497 TI - Syncopal episodes in a young amateur body builder. AB - A 36 year old male weight training enthusiast suffered several syncopal episodes. An electrocardiogram confirmed atrial fibrillation with normal ventricular response. The patient admitted to taking anabolic steroids and bromocriptine. The atrial fibrillation was considered to be due to bromocriptine misuse. PMID- 11867496 TI - Two cases of peroneus brevis tendon tear. AB - A longitudinal tear of the peroneal tendon is thought to be the result of repetitive peroneal subluxation. However, this report documents two cases of longitudinal split of the peroneus brevis tendon that had no peroneal tendon subluxation. Primary suture was performed. Subluxation of the peroneal tendons was not identified surgically in either case. PMID- 11867498 TI - Haemodynamics during a complete exercise induced atrioventricular block. AB - The case is reported of an athlete who, during an exercise test, had a complete atrioventricular block without symptoms of cardiac output failure. Heart rate, stroke index, cardiac index, and myocardial contractility were monitored during the episode by an impedance cardiograph. The most important findings of this report are the continuous increase in stroke index, which compensated for the lack of heart rate response, and the normal cardiac index values achieved during the exercise. This stroke index response was mainly due to an increase in myocardial contractility. PMID- 11867499 TI - Osteomyelitis pubis versus osteitis pubis: a case presentation and review of the literature. AB - An athletic 23 year old man presented with suprapubic tenderness, fever, and raised inflammatory blood variables. A diagnostic laparoscopy was performed, with a presumed diagnosis of retrocaecal appendicitis, but no abnormalities were found, apart from free fluid in the pouch of Douglas. Imaging of the pubic area suggested bony infection and inflammation. Biopsy and culture confirmed the presence of Staphylococcus aureus, a very common pathogen. The final diagnosis was osteomyelitis pubis, an infectious disease, and osteitis pubis, an inflammatory disease. PMID- 11867500 TI - Diastasis of the pubic symphysis peculiar to horse riders: modern aspects of pelvic pommel injuries. AB - Diastasis of the pubic symphysis is a well documented injury typically associated with high energy trauma. Three cases in horse riders are here described, emphasising the appropriate modern investigation, including computed tomography, and orthopaedic and urological management. PMID- 11867501 TI - The "piriformis syndrome"--myth or reality? PMID- 11867502 TI - Oral cancer risk perception among participants in an oral cancer screening program. AB - Oral cancer screening provides an opportunity for early detection and for education to high-risk tobacco and alcohol users. To plan interventions that would motivate oral cancer screening and risk reduction, we surveyed oral cancer risk perception and risk behaviors among participants in a free oral-cancer screening. Participants (N = 803) were racially diverse; 43% had a history of smoking and 9%, a history of alcohol abuse. Current smokers and those with higher lifetime tobacco exposure perceived themselves to be at higher risk for cancer than nonsmokers or those with less lifetime tobacco exposure (all ps, <0.01). Alcohol use was unrelated to oral cancer risk perception (p > 0.05). Compared with women, men (p = 0.01) felt more at risk; compared with other racial groups, Asians (p < 0.05) felt less at risk. Demographic differences were explained by differences in risk behaviors. Current smoking status (beta, 0.196; p < 0.001) and level of tobacco exposure (beta, 0.100; p < 0.05) were the only significant independent predictors of heightened risk perception (R(2) = 0.09). Those with alcohol abuse histories, and older smokers, may need increased education. These findings support the need for health education materials that incorporate the oral cancer risk perception of high-risk individuals. PMID- 11867503 TI - Generating longitudinal screening algorithms using novel biomarkers for disease. AB - Recent advances in molecular technology are leading to the discovery of new tumor biomarkers that may be useful for cancer screening and early diagnosis. Translating a potential screening biomarker from the laboratory to its use in patient care may require an algorithm or screening rule for its application. An algorithm that can detect the smallest deviation from a defined norm is likely to achieve the highest sensitivity, but any practical screening algorithm must do so with strict controls on test specificity to avoid false-positive results, and unnecessary patient alarm and risk. Longitudinal algorithms that make use of previous tumor marker values and trends are likely to obtain improvements over single threshold rules. Thus far, a few longitudinal screening algorithms have been proposed (e.g., using serial prostate-specific antigen values for the detection of prostate cancer and serial CA125 values for the detection of ovarian cancer), but these algorithms are not appropriate for novel tumor marker discoveries, because they rely on unverifiable assumptions that may not translate to the behavior of the new marker. The algorithm presented here is motivated by: (a) the need to develop an algorithm for early detection using novel markers; (b) the practical demands on data and specimen availability; and (c) the need to be robust enough to accommodate a wide range of tumor growth behavior. We use Parametric Empirical Bayes statistical theory to model the trajectory of markers over time in a cohort of asymptomatic healthy subjects, and use the estimated trajectory to produce person-specific thresholds that depend on the screening history of each person. The thresholds are chosen to give the person (or population) a specified false-positive rate. The resulting algorithm is simple and can be represented in a simple graph or a chart. The statistical analysis needed to generate the algorithm can be found in nearly every basic statistical package. The algorithm is highly robust and can detect a wide range of tumor behaviors. The Parametric Empirical Bayes screening algorithm should take a central role when evaluating marker discoveries for use in screening. The algorithm is particularly useful when screening with a new marker of which the behavior in the preclinical period is not well known. PMID- 11867505 TI - Diagnostic X-rays and ultrasound exposure and risk of childhood acute lymphoblastic leukemia by immunophenotype. AB - The objective of this study was to evaluate the association between in utero diagnostic X-rays and childhood acute lymphoblastic leukemia (ALL) and the less well-studied relationship of this malignancy to preconception and postnatal diagnostic X-rays or fetal ultrasound exposures. The Children's Cancer Group conducted a case-control study including interviews with parents of 1842 ALL cases diagnosed under the age of 15 years and 1986 individually matched controls. Associations of self-reported parental preconception, in utero, and postnatal X ray exposure with risk of childhood ALL were examined using odds ratios (ORs) and corresponding 95% confidence intervals (CIs) obtained from logistic regression models among the overall group of ALL cases as well as immunophenotypic and age specific subgroups. Overall, in utero pelvimetric diagnostic X-rays were not associated with the risk of pediatric ALL (OR, 1.2; 95% CI, 0.8-1.7). Childhood ALL, all types combined (OR, 1.1; 95% CI, 0.9-1.2) and specific types were also not linked with postnatal diagnostic X-ray exposures. Neither maternal (OR, 0.9; 95% CI, 0.8-1.2) nor paternal (OR, 1.1; 95% CI, 0.8-1.4) lower abdominal preconception diagnostic X-rays were associated with risk of childhood ALL. Among the multiple comparisons for age-, sex-, and subtype-specific subgroups, we observed an elevated risk of total ALL among children ages 11-14 at diagnosis (OR, 2.4; 95% CI, 1.1-5.0) in relation to in utero pelvimetric diagnostic X-ray exposures and a small increase in pre-B ALL for all ages combined (OR, 1.7; 95% CI, 1.1-2.7) in relation to postnatal diagnostic X-rays. In utero diagnostic ultrasound tests were not linked with risk of childhood ALL. We found little consistent evidence that in utero diagnostic ultrasound tests or X-rays were linked with an increased risk of childhood ALL. Small increases in total or pre-B ALL risks for children in selected age groups to very low ionizing radiation exposures from postnatal or preconception diagnostic X-ray exposures may represent chance findings or biases. Future studies of diagnostic X-rays and childhood leukemia in the United States will require extensive additional efforts and resources to quantify risk because of declining in utero exposures in the general population (thus necessitating large numbers of subjects, particularly cases) and the difficulty in validating reported exposures. PMID- 11867506 TI - Carbohydrates and colorectal cancer risk among Chinese in North America. AB - Previous studies have analyzed total carbohydrate as a dietary risk factor for colorectal cancer (CRC) but obtained conflicting results, perhaps attributable in part to the embedded potential confounder, fiber. The aim of this study was to analyze the nonfiber ("effective") carbohydrate component (eCarb) separately and to test the hypothesis that effective carbohydrate consumption is directly related to CRC risk. The data (473 cases and 1192 controls) were from a large, multicenter, case-control study of Chinese residing in North America. Multivariate logistic regression was used to perform a secondary analysis controlling for age; sex; consumption of fat, fiber, calcium, and total kilocalories; body mass (Quetelet's) index; family history; education; and years in North America. Odds ratios (ORs) and 95% confidence intervals (CIs) were calculated to estimate risk among subgroups by sex and cancer site. A statistically significant positive association was observed between eCarb consumption and risk of CRC in both men (OR, 1.7 comparing highest with lowest tertile of eCarb consumption; 95% CI, 1.1-2.7) and women (OR, 2.7; 95% CI, 1.5 4.8). As expected, the ORs for total carbohydrate were somewhat lower than those for effective carbohydrate, but the differences were not large. A sex difference in risk by colorectal subsite was observed, with risk concentrated in the right colon for women (OR, 6.5; 95% CI, 2.4-18.4) and in the rectum for men (OR, 2.4; 95% CI, 1.2-4.8). These data indicate that increased eCarb and total carbohydrate consumption are both associated with increased risk of CRC in both sexes, and that among women, relative risk appears greatest for the right colon, whereas among men, relative risk appears greatest for the rectum. PMID- 11867504 TI - Effects of oral administration of N-acetyl-L-cysteine: a multi-biomarker study in smokers. AB - N-Acetyl-L-cysteine (NAC) has been shown to exert cancer-protective mechanisms and effects in experimental models. We report here the results of a randomized, double-blind, placebo-controlled, Phase II chemoprevention trial with NAC in healthy smoking volunteers. The subjects were supplemented daily with 2 x 600 mg of oral tablets of NAC (n = 20) or placebo (n = 21) for a period of 6 months, and internal dose markers [plasma and bronchoalveolar lavage (BAL) fluid cotinine, urine mutagenicity], biologically effective dose markers [smoking-related DNA adducts and hemoglobin (Hb) adducts], and biological response markers (micronuclei frequency and antioxidants scavenging capacity) were assessed at both pre- and postsupplementation times (T(0) and T(1), respectively). Overall, the internal dose markers remained unchanged at T(1) as compared with T(0) in both NAC and placebo groups. When quantifying the biologically effective dose markers, we observed an inhibitory effect of NAC toward the formation of lipophilic-DNA adducts (5.18 +/- 0.73 versus 4.08 +/- 1.03/10(8) nucleotides; mean +/- SE; P = 0.05) as well as of 7,8-dihydro-8-oxo-2'-deoxyguanosine adducts in BAL cells (3.9 +/- 0.6 versus 2.3 +/- 0.2/10(5) nucleotides; P = 0.003). There was no effect of NAC on the formation of lipophilic-DNA adducts in peripheral blood lymphocytes or polycyclic aromatic hydrocarbon-DNA adducts in mouth floor/buccal mucosa cells or 4-aminobiphenyl-Hb adducts. Likewise, quantification of the biological response markers showed an inhibitory effect of NAC on the frequency of micronuclei in mouth floor and in soft palate cells (1.3 +/- 0.2 versus 0.9 +/- 0.2; P = 0.001) and a stimulating effect of NAC on plasma antioxidant scavenging capacity (393 +/- 14 versus 473 +/- 19 microM Trolox; P = 0.1) but not on BAL fluid antioxidant scavenging capacity. We conclude that NAC has the potential to impact upon tobacco smoke carcinogenicity in humans because it can modulate certain cancer-associated biomarkers in specific organs. PMID- 11867507 TI - A randomized isoflavone intervention among premenopausal women. AB - Isoflavones, phytoestrogens contained in soy foods, may play a role in breast cancer prevention. This randomized double-blinded trial with 34 premenopausal women investigated whether 100 mg of isoflavones per day versus placebo affects the ovulatory cycle during 1 year. Compliance with the study regimen was confirmed by the increase of urinary isoflavone excretion among the intervention group. Blood samples were taken 5 days after ovulation as determined by an ovulation kit, at baseline, and at months 1, 3, 6, and 12. Serum levels of estrone, estradiol, estrone sulfate, progesterone, sex hormone-binding globulin, follicle-stimulating hormone, and luteinizing hormone were quantified by immunoassay; free estradiol was calculated. We applied the method of least squares to fit general linear models to test for an intervention effect while taking into account the repeated measurement design. Except for a small difference in age, the two groups were comparable at baseline. Menstrual cycle length did not change significantly during the intervention [F(1,32) = 0.69; P = 0.44]. During 1 year, we did not observe any significant changes in hormone levels by treatment group. The difference in change between intervention and control group was -13.0 pg/ml (95% confidence interval, -57.5 to 31.5) for estradiol and 6.9 pg/ml (95% confidence interval, -17.8 to 31.5) for estrone. Exclusion of 22 non-ovulatory cycles, noncompliant women, or non-Asian women did not affect the results. These findings do not support the hypothesis that isoflavones affect the ovulatory cycles of premenopausal women over a 1-year period. However, isoflavones alone may have different effects on the reproductive cycle than isoflavones present in soy foods. PMID- 11867508 TI - Increasing fruit and vegetable intake among adults attending colorectal cancer screening: the efficacy of a brief tailored intervention. AB - Fruits and vegetables appear to confer protection against several cancers, but most adults in the United Kingdom eat substantially less than the recommended amounts. Cancer screening services could provide a valuable context in which to provide advice on increasing fruit and vegetable intake. This study examined the efficacy of a brief, tailored, psycho-educational intervention for increasing fruit and vegetable intake, carried out in a cancer screening clinic. The study was a randomized, controlled trial. 742 participants, 55-64 years of age, recruited from three cancer screening clinics, completed a baseline questionnaire. They were assigned either to the tailored intervention group or to an untreated control group. The primary outcome measure was self-reported consumption of fruit and vegetables. At 6 week follow-up there were significant increases in daily servings of fruit and vegetables in the tailored intervention group (CI, 0.87-1.25) compared with the untreated group (CI, 0.08-0.43). These results support the efficacy of a simple, written message, which is tailored to the intake and knowledge levels of the individual, for modifying cancer protective dietary behaviors, at least in the short term. They also suggest that cancer screening clinics may be a good context for providing this service. PMID- 11867509 TI - Early life factors in relation to breast cancer risk in postmenopausal women. AB - We evaluated the role of early life factors in a large, population-based, case control study of breast cancer risk in postmenopausal women. Case women in Massachusetts, New Hampshire, and Wisconsin were ascertained through state cancer registries; control women were randomly selected from drivers license lists (50 65 years of age) or Medicare beneficiary lists (65-79 years of age). Information concerning factors of interest was obtained through structured telephone interviews. Overall, 83% of eligible cases and 78% of eligible controls participated, and data from more than 2900 women were available for this analysis. We observed a weak J-shaped relationship between birth weight and breast cancer risk; the increased risk was not statistically significant for either the lowest or the highest birth weight. Parental smoking during the pregnancy was not associated with risk of breast cancer in the adult daughter. Breast cancer risk increased significantly with father's education (P = 0.01). Risk also increased with greater age of the mother at the time of the subject's birth (P = 0.04). The subject's birth rank was inversely associated with risk (P = 0.03), as was the number of older sisters (P = 0.03), but the number of older brothers, number of younger siblings, sibship gender ratio, and total sibship size were unrelated to risk. Overall, our results are consistent with previous studies and suggest that these early life factors have a modest influence on breast cancer risk in postmenopausal women. PMID- 11867510 TI - Transforming growth factor beta receptor 1 polyalanine polymorphism and exon 5 mutation analysis in breast and ovarian cancer. PMID- 11867511 TI - No association between a single nucleotide polymorphism in CYP19 and breast cancer risk. PMID- 11867512 TI - Correspondence re: Terry et al., No association between fat and fatty acids intake and risk of colorectal cancer. Cancer Epidemiol. Biomark. Prev., 10: 913 914, 2001. PMID- 11867513 TI - Correspondence re: Weinberg et al., Cholecystokinin and gastrin levels are not elevated in pancreatic carcinoma. Cancer Epidemiol. Biomark. Prev., 10: 721-722, 2001. PMID- 11867515 TI - H5 avian and H9 swine influenza virus haemagglutinin structures: possible origin of influenza subtypes. AB - There are 15 subtypes of influenza A virus (H1-H15), all of which are found in avian species. Three caused pandemics in the last century: H1 in 1918 (and 1977), H2 in 1957 and H3 in 1968. In 1997, an H5 avian virus and in 1999 an H9 virus caused outbreaks of respiratory disease in Hong Kong. We have determined the three-dimensional structures of the haemagglutinins (HAs) from H5 avian and H9 swine viruses closely related to the viruses isolated from humans in Hong Kong. We have compared them with known structures of the H3 HA from the virus that caused the 1968 H3 pandemic and of the HA--esterase--fusion (HEF) glycoprotein from an influenza C virus. Structure and sequence comparisons suggest that HA subtypes may have originated by diversification of properties that affected the metastability of HAs required for their membrane fusion activities in viral infection. PMID- 11867514 TI - On the genetics of retinitis pigmentosa and on mutation-independent approaches to therapeutic intervention. AB - Retinitis pigmentosa (RP), the group of hereditary conditions involving death of retinal photoreceptors, represents the most prevalent cause of visual handicap among working populations in developed countries. Here we provide an overview of the molecular pathologies associated with such disorders, from which it becomes clearly apparent that RP is one of the most genetically heterogeneous of hereditary conditions for which molecular pathologies have so far been elucidated. While heterogeneity of such magnitude would appear to represent a major impediment to the development of therapeutics, mutation-independent approaches to therapy are being developed to effectively by-pass such diversity in genetic aetiology. The implications of such technologies in terms of therapeutic intervention in RP, and indeed other genetically heterogeneous conditions, will be addressed. PMID- 11867516 TI - The morphogenic linker peptide of HBV capsid protein forms a mobile array on the interior surface. AB - Many capsid proteins have peptides that influence their assembly. In hepatitis B virus capsid protein, the peptide STLPETTVV, linking the shell-forming 'core' domain and the nucleic acid-binding 'protamine' domain, has such a role. We have studied its morphogenic properties by permuting its sequence, substituting it with an extraneous peptide, deleting it to directly fuse the core and protamine domains and assembling core domain dimers with added linker peptides. The peptide was found to be necessary for the assembly of protamine domain-containing capsids, although its size-determining effect tolerates some modifications. Although largely invisible in a capsid crystal structure, we could visualize linker peptides by cryo-EM difference imaging: they emerge on the inner surface and extend from the capsid protein dimer interface towards the adjacent symmetry axis. A closely sequence-similar peptide in cellobiose dehydrogenase, which has an extended conformation, offers a plausible prototype. We propose that linker peptides are attached to the capsid inner surface as hinged struts, forming a mobile array, an arrangement with implications for morphogenesis and the management of encapsidated nucleic acid. PMID- 11867517 TI - The rhesus rotavirus VP4 sialic acid binding domain has a galectin fold with a novel carbohydrate binding site. AB - Cell attachment and membrane penetration are functions of the rotavirus outer capsid spike protein, VP4. An activating tryptic cleavage of VP4 produces the N terminal fragment, VP8*, which is the viral hemagglutinin and an important target of neutralizing antibodies. We have determined, by X-ray crystallography, the atomic structure of the VP8* core bound to sialic acid and, by NMR spectroscopy, the structure of the unliganded VP8* core. The domain has the beta-sandwich fold of the galectins, a family of sugar binding proteins. The surface corresponding to the galectin carbohydrate binding site is blocked, and rotavirus VP8* instead binds sialic acid in a shallow groove between its two beta-sheets. There appears to be a small induced fit on binding. The residues that contact sialic acid are conserved in sialic acid-dependent rotavirus strains. Neutralization escape mutations are widely distributed over the VP8* surface and cluster in four epitopes. From the fit of the VP8* core into the virion spikes, we propose that VP4 arose from the insertion of a host carbohydrate binding domain into a viral membrane interaction protein. PMID- 11867518 TI - SHEPHERD is the Arabidopsis GRP94 responsible for the formation of functional CLAVATA proteins. AB - The Arabidopsis shepherd (shd) mutant shows expanded shoot apical meristems (SAM) and floral meristems (FM), disorganized root apical meristems, and defects in pollen tube elongation. We have discovered that SHD encodes an ortholog of GRP94, an ER-resident HSP90-like protein. Since the shd phenotypes in SAM and FM are similar to those of the clavata (clv) mutants, we have explored the possibility that CLV complex members could be SHD targets. The SAM and FM morphology of shd clv double mutants are indistinguishable from those of clv single mutants, and the wuschel (wus) mutation is completely epistatic to the shd mutation, indicating that SHD and CLV act in the same genetic pathway to suppress WUS function. Moreover, the effects of CLV3 overexpression that result in the elimination of SAM activity were abolished in the shd mutant, indicating that CLV function is dependent on SHD function. Therefore, we conclude that the SHD protein is required for the correct folding and/or complex formation of CLV proteins. PMID- 11867520 TI - The Ppz protein phosphatases are key regulators of K+ and pH homeostasis: implications for salt tolerance, cell wall integrity and cell cycle progression. AB - The yeast Ppz protein phosphatases and the Hal3p inhibitory subunit are important determinants of salt tolerance, cell wall integrity and cell cycle progression. We present several lines of evidence showing that these disparate phenotypes are connected by the fact that Ppz regulates K+ transport. First, salt tolerance, cell wall integrity and cell cycle phenotypes of Ppz mutants are dependent on the Trk K+ transporters. Secondly, Ppz mutants exhibit altered activity of the Trk system, as measured by rubidium uptake. Thirdly, Ppz mutants exhibit altered intracellular K+ and pH, as expected from H+ efflux providing electrical balance during K+ uptake. Our unifying picture of Ppz phenotypes contends that activation of Trk by decreased Ppz activity results in plasma membrane depolarization (reducing uptake of toxic cations), increased intracellular K+ and turgor (compromising cell integrity), and increased intracellular pH (augmenting the expression of pH-regulated genes and facilitating alpha-factor recovery). In addition to providing a coherent explanation for all Ppz-dependent phenotypes, our results provide evidence for a causal relationship between intracellular cation homeostasis and a potential cell cycle checkpoint. PMID- 11867519 TI - Transformation of local Ca2+ spikes to global Ca2+ transients: the combinatorial roles of multiple Ca2+ releasing messengers. AB - In pancreatic acinar cells, low, threshold concentrations of acetylcholine (ACh) or cholecystokinin (CCK) induce repetitive local cytosolic Ca2+ spikes in the apical pole, while higher concentrations elicit global signals. We have investigated the process that transforms local Ca2+ spikes to global Ca2+ transients, focusing on the interactions of multiple intracellular messengers. ACh-elicited local Ca2+ spikes were transformed into a global sustained Ca2+ response by cyclic ADP-ribose (cADPR) or nicotinic acid adenine dinucleotide phosphate (NAADP), whereas inositol 1,4,5-trisphosphate (IP3) had a much weaker effect. In contrast, the response elicited by a low CCK concentration was strongly potentiated by IP3, whereas cADPR and NAADP had little effect. Experiments with messenger mixtures revealed a local interaction between IP3 and NAADP and a stronger global potentiating interaction between cADPR and NAADP. NAADP strongly amplified the local Ca2+ release evoked by a cADPR/IP3 mixture eliciting a vigorous global Ca2+ response. Different combinations of Ca2+ releasing messengers can shape the spatio-temporal patterns of cytosolic Ca2+ signals. NAADP and cADPR are emerging as key messengers in the globalization of Ca2+ signals. PMID- 11867521 TI - C/EBPalpha triggers proteasome-dependent degradation of cdk4 during growth arrest. AB - CCAAT/enhancer binding protein alpha (C/EBPalpha) causes growth arrest via direct interaction with the cyclin-dependent kinases cdk2 and cdk4. In this paper, we present evidence showing that C/EBPalpha enhances a proteasome-dependent degradation of cdk4 during growth arrest in liver of newborn mice and in cultured cells. Overexpression of C/EBPalpha in several biological systems leads to a reduction of cdk4 protein levels, but not mRNA levels. Experiments with several tissue culture models reveal that C/EBPalpha enhances the formation of cdk4 ubiquitin conjugates and induces degradation of cdk4 through a proteasome dependent pathway. As a result, the half-life of cdk4 is shorter and protein levels of cdk4 are reduced in cells expressing C/EBPalpha. Gel filtration analysis of cdk4 complexes shows that a chaperone complex cdk4-cdc37-Hsp90, which protects cdk4 from degradation, is abundant in proliferating livers that lack C/EBPalpha, but this complex is weak or undetectable in livers expressing C/EBPalpha. Our studies show that C/EBPalpha disrupts the cdk4-cdc37-Hsp90 complex via direct interaction with cdk4 and reduces protein levels of cdk4 by increasing proteasome-dependent degradation of cdk4. PMID- 11867522 TI - The Tim9p-Tim10p complex binds to the transmembrane domains of the ADP/ATP carrier. AB - The soluble Tim9p-Tim10p (Tim, translocase of inner membrane) complex of the mitochondrial intermembrane space mediates the import of the carrier proteins and is a component of the TIM22 import system. The mechanism by which the Tim9p Tim10p complex assembles and binds the carriers is not well understood, but previous studies have proposed that the conserved cysteine residues in the 'twin CX3C' motif coordinate zinc and potentially generate a zinc-finger-like structure that binds to the matrix loops of the carrier proteins. Here we have purified the native and recombinant Tim9p-Tim10p complex, and show that both complexes resemble each other and consist of three Tim9p and three Tim10p. Results from inductively coupled plasma--mass spectrometry studies failed to detect zinc in the Tim9p-Tim10p complex. Instead, the cysteine residues seemingly formed disulfide linkages. The Tim9p-Tim10p complex bound specifically to the transmembrane domains of the ADP/ATP carrier, but had no affinity for Tim23p, an inner membrane protein that is inserted via the TIM22 complex. The chaperone-like Tim9p-Tim10p complex thus may prevent aggregation of the unfolded carrier proteins in the aqueous intermembrane space. PMID- 11867523 TI - Epstein-Barr virus RNA confers resistance to interferon-alpha-induced apoptosis in Burkitt's lymphoma. AB - We investigated whether Epstein--Barr virus (EBV) infection could counteract the antitumor effect of interferon (IFN)-alpha. EBV-negative subclones isolated from EBV-positive Burkitt's lymphoma (BL) cell lines Akata, Daudi and Mutu were found to fall into apoptosis after IFN-alpha treatment. On the other hand, EBV-positive counterparts exhibited striking resistance against IFN-alpha-induced apoptosis. Transfection of an individual EBV latent gene into EBV-negative BL cells revealed that EBV-encoded poly(A)(-) RNAs (EBERs) were responsible for IFN resistance. EBERs bound double-stranded (ds) RNA-activated protein kinase (PKR), a key mediator of the antiviral effect of IFN-alpha, and inhibited its phosphorylation. Transfection of dominant-negative PKR, which was catalytically inactive and could block phosphorylation of endogenous PKR, made EBV-negative BL cells resistant to IFN-alpha-induced apoptosis. Furthermore, EBERs did not bind mutant PKR, which was catalytically active but lacked dsRNA-binding activity, nor did they inhibit its phosphorylation. These results indicate that EBERs confer resistance to IFN alpha-induced apoptosis via binding to PKR and inhibition of its phosphorylation. This is the first report that the virus counteracts IFN-induced apoptosis in virus-associated tumors. PMID- 11867524 TI - The Wnt antagonist Dickkopf-1 is regulated by Bmp signaling and c-Jun and modulates programmed cell death. AB - Dickkopf-1 (Dkk-1) has been shown to be a potent inhibitor of Wnt/beta-catenin signaling in a variety of assays and organisms. In this study, we show that expression of Dkk-1 overlaps significantly with the sites of programmed cell death in normal as well as mutant vertebrate limb development, and identify several of its upstream regulators, one of which is Bmp-4. Interestingly, Bmp-4 only activates Dkk-1 when it concomitantly induces apoptosis. Moreover, Dkk-1 is heavily up-regulated by UV irradiation and several other genotoxic stimuli. We further show that normal expression of Dkk-1 is dependent on the Ap-1 family member c-Jun and that overexpression of Dkk-1 enhances Bmp-triggered apoptosis in the vertebrate limb. Taken together, our results provide evidence for an important role of Dkk-1-mediated inhibition of Wnt/beta-catenin signaling in response to different stress signals that all converge on the activation of c-Jun in vivo. PMID- 11867525 TI - The planar polarity gene strabismus regulates convergent extension movements in Xenopus. AB - The signaling mechanisms that specify, guide and coordinate cell behavior during embryonic morphogenesis are poorly understood. We report that a Xenopus homolog of the Drosophila planar cell polarity gene strabismus (stbm) participates in the regulation of convergent extension, a critical morphogenetic process required for the elongation of dorsal structures in vertebrate embryos. Overexpression of Xstbm, which is expressed broadly in early development and subsequently in the nervous system, causes severely shortened trunk structures; a similar phenotype results from inhibiting Xstbm translation using a morpholino antisense oligo. Experiments with Keller explants further demonstrate that Xstbm can regulate convergent extension in both dorsal mesoderm and neural tissue. The specification of dorsal tissues is not affected. The Xstbm phenotype resembles those obtained with several other molecules with roles in planar polarity signaling, including Dishevelled and Frizzled-7 and -8. Unlike these proteins, however, Stbm has little effect on conventional Wnt/beta-catenin signaling in either frog or fly assays. Thus our results strongly support the emerging hypothesis that a vertebrate analog of the planar polarity pathway governs convergent extension movements. PMID- 11867526 TI - The G-protein gamma subunit gpc-1 of the nematode C.elegans is involved in taste adaptation. AB - Caenorhabditis elegans has two heterotrimeric G-protein gamma subunits, gpc-1 and gpc-2. Although GPC-1 is specifically expressed in sensory neurons, it is not essential for the detection of odorants or salts. To test whether GPC-1 is involved in sensory plasticity, we developed a water soluble compound adaptation assay. The behaviour of wild-type animals in this assay confirms that prolonged exposure to salts can abolish chemo-attraction to these compounds. This process is time and concentration dependent, partly salt specific and reversible. In contrast, gpc-1 mutant animals show clear deficits in their ability to adapt to NaAc, NaCl and NH4Cl, but normal wild-type adaptation to odorants. Two other loci previously implicated in odorant adaptation, adp-1 and osm-9, are also involved in adaptation to salts. Our finding that G proteins, OSM-9 and ADP-1 are involved in taste adaptation offer the first molecular insight into this process. PMID- 11867527 TI - The SecYEG preprotein translocation channel is a conformationally dynamic and dimeric structure. AB - Escherichia coli preprotein translocase comprises a membrane-embedded trimeric complex of SecY, SecE and SecG. Previous studies have shown that this complex forms ring-like assemblies, which are thought to represent the preprotein translocation channel across the membrane. We have analyzed the functional state and the quaternary structure of the SecYEG translocase by employing cross-linking and blue native gel electrophoresis. The results show that the SecYEG monomer is a highly dynamic structure, spontaneously and reversibly associating into dimers. SecG-dependent tetramers and higher order SecYEG multimers can also exist in the membrane, but these structures form at high SecYEG concentration or upon overproduction of the complex only. The translocation process does not affect the oligomeric state of the translocase and arrested preproteins can be trapped with SecYEG or SecYE dimers. Dissociation of the dimer into a monomer by detergent induces release of the trapped preprotein. These results provide direct evidence that preproteins cross the bacterial membrane, associated with a translocation channel formed by a dimer of SecYEG. PMID- 11867528 TI - Role of the PLC-related, catalytically inactive protein p130 in GABA(A) receptor function. AB - The protein p130 was isolated from rat brain as an inositol 1,4,5-trisphosphate binding protein with a domain organization similar to that of phospholipase C delta1 but lacking PLC activity. We show that p130 plays an important role in signaling by the type A receptor for gamma-aminobutyric acid (GABA). Yeast twohybrid screening identified GABARAP (GABA(A) receptor-associated protein), which is proposed to contribute to the sorting, targeting or clustering of GABA(A) receptors, as a protein that interacts with p130. Furthermore, p130 competitively inhibited the binding of the gamma2 subunit of the GABA(A) receptor to GABARAP in vitro. Electrophysiological analysis revealed that the modulation of GABA-induced Cl- current by Zn2+ or diazepam, both of which act at GABA(A) receptors containing gamma subunits, is impaired in hippocampal neurons of p130 knockout mice. Moreover, behavioral analysis revealed that motor coordination was impaired and the intraperitoneal injection of diazepam induced markedly reduced sedative and antianxiety effects in the mutant mice. These results indicate that p130 is essential for the function of GABA(A) receptors, especially in response to the agents acting on a gamma2 subunit. PMID- 11867529 TI - The C. elegans ric-3 gene is required for maturation of nicotinic acetylcholine receptors. AB - Mutations in ric-3 (resistant to inhibitors of cholinesterase) suppress the neuronal degenerations caused by a gain of function mutation in the Caenorhabditis elegans DEG-3 acetylcholine receptor. RIC-3 is a novel protein with two transmembrane domains and extensive coiled-coil domains. It is expressed in both muscles and neurons, and the protein is concentrated within the cell bodies. We demonstrate that RIC-3 is required for the function of at least four nicotinic acetylcholine receptors. However, GABA and glutamate receptors expressed in the same cells are unaffected. In ric-3 mutants, the DEG-3 receptor accumulates in the cell body instead of in the cell processes. Moreover, co expression of ric-3 in Xenopus laevis oocytes enhances the activity of the C.elegans DEG-3/DES-2 and of the rat alpha-7 acetylcholine receptors. Together, these data suggest that RIC-3 is specifically required for the maturation of acetylcholine receptors. PMID- 11867531 TI - Conversion of raft associated prion protein to the protease-resistant state requires insertion of PrP-res (PrP(Sc)) into contiguous membranes. AB - Prion protein (PrP) is usually attached to membranes by a glycosylphosphatidylinositol-anchor that associates with detergent-resistant membranes (DRMs), or rafts. To model the molecular processes that might occur during the initial infection of cells with exogenous transmissible spongiform encephalopathy (TSE) agents, we examined the effect of membrane association on the conversion of the normal protease-sensitive PrP isoform (PrP-sen) to the protease-resistant isoform (PrP-res). A cell-free conversion reaction approximating physiological conditions was used, which contained purified DRMs as a source of PrP-sen and brain microsomes from scrapie-infected mice as a source of PrP-res. Interestingly, DRM-associated PrP-sen was not converted to PrP-res until the PrP-sen was either released from DRMs by treatment with phosphatidylinositol-specific phospholipase C (PI-PLC), or the combined membrane fractions were treated with the membrane-fusing agent polyethylene glycol (PEG). PEG-assisted conversion was optimal at pH 6--7, and acid pre-treating the DRMs was not sufficient to permit conversion without PI-PLC or PEG, arguing against late endosomes/lysosomes as primary compartments for PrP conversion. These observations raise the possibility that generation of new PrP-res during TSE infection requires (i) removal of PrP-sen from target cells; (ii) an exchange of membranes between cells; or (iii) insertion of incoming PrP-res into the raft domains of recipient cells. PMID- 11867530 TI - K-Ras mediates cytokine-induced formation of E-cadherin-based adherens junctions during liver development. AB - The E-cadherin-based adherens junction (AJ) is essential for organogenesis of epithelial tissues including the liver, although the regulatory mechanism of AJ formation during development remains unknown. Using a primary culture system of fetal hepatocytes in which oncostatin M (OSM) induces differentiation, we show here that OSM induces AJ formation by altering the subcellular localization of AJ components including E-cadherin and catenins. By retroviral expression of dominant-negative forms of signaling molecules, Ras was shown to be required for the OSM-induced AJ formation. Fetal hepatocytes derived from K-Ras knockout (K Ras-/-) mice failed to form AJs in response to OSM, whereas AJ formation was induced normally by OSM in mutant hepatocytes lacking both H-Ras and N-Ras. Moreover, the defective phenotype of K-Ras-/- hepatocytes was restored by expression of K-Ras, but not by H-Ras and N-Ras. Finally, pull-down assays using the Ras-binding domain of Raf1 demonstrated that OSM directly activates K-Ras in fetal hepatocytes. These results indicate that K-Ras specifically mediates cytokine signaling for formation of AJs during liver development. PMID- 11867533 TI - Structure of a functional IGF2R fragment determined from the anomalous scattering of sulfur. AB - Insulin-like growth factor II receptor (IGF2R) is a multifunctional cell surface receptor implicated in tumour suppression. Its growth inhibitory activity has been associated with an ability to bind IGF-II. IGF2R contains 15 homologous extracellular domains, with domain 11 primarily responsible for IGF-II binding. We report a 1.4 A resolution crystal structure of domain 11, solved using the anomalous scattering signal of sulfur. The structure consists of two crossed beta sheets forming a flattened beta-barrel. Structural analysis identifies the putative IGF-II binding site at one end of the beta-barrel whilst crystal lattice contacts suggest a model for the full-length IGF2R extracellular region. The structure factors and coordinates of IGF2R domain 11 have been deposited in the Protein Data Bank (accession codes 1GP0 and 1GP3). PMID- 11867532 TI - Visualization of the ER-to-cytosol dislocation reaction of a type I membrane protein. AB - The human cytomegalovirus gene products US2 and US11 induce proteasomal degradation of MHC class I heavy chains. We have generated an enhanced green fluorescent protein-class I heavy chain (EGFP-HC) chimeric molecule to study its dislocation and degradation in US2- and US11-expressing cells. The EGFP-HC fusion is stable in control cells, but is degraded rapidly in US2- or US11-expressing cells. Proteasome inhibitors induce in a time-dependent manner the accumulation of EGFP-HC molecules in US2- and US11-expressing cells, as assessed biochemically and by cytofluorimetry of intact cells. Pulse-chase analysis and subcellular fractionation show that EGFP-HC proteins are dislocated from the endoplasmic reticulum and can be recovered as deglycosylated fluorescent intermediates in the cytosol. These results raise the possibility that dislocation of glycoproteins from the ER may not require their full unfolding. PMID- 11867535 TI - Drosophila Gp150 is required for early ommatidial development through modulation of Notch signaling. AB - Cellular signaling activities must be tightly regulated for proper cell fate control and tissue morphogenesis. Here we report that the Drosophila leucine-rich repeat transmembrane glycoprotein Gp150 is required for viability, fertility and development of the eye, wing and sensory organs. In the eye, Gp150 plays a critical role in regulating early ommatidial formation. Gp150 is highly expressed in cells of the morphogenetic furrow (MF) region, where it accumulates exclusively in intracellular vesicles in an endocytosis-independent manner. Loss of gp150 function causes defects in the refinement of photoreceptor R8 cells and recruitment of other cells, which leads to the formation of aberrant ommatidia. Genetic analyses suggest that Gp150 functions to modulate Notch signaling. Consistent with this notion, Gp150 is co-localized with Delta in intracellular vesicles in cells within the MF region and loss of gp150 function causes accumulation of intracellular Delta protein. Therefore, Gp150 might function in intracellular vesicles to modulate Delta-Notch signaling for cell fate control and tissue morphogenesis. PMID- 11867534 TI - A Caenorhabditis elegans TGF-beta, DBL-1, controls the expression of LON-1, a PR related protein, that regulates polyploidization and body length. AB - Using cDNA-based array analysis combined with double-stranded RNA interference (dsRNAi), we have identified yk298h6 as a target gene of Caenorhabditis elegans TGF-beta signaling. Worms overexpressing dbl-1, a TGF-beta ligand, are 16% longer than wild type. Array analysis shows yk298h6 to be one of several genes suppressed in such worms. Disruption of yk298h6 function by dsRNAi also resulted in long worms, suggesting that it is a negative regulator of body length. yk298h6 was then mapped to, and shown to be identical to, lon-1, a known gene that affects body length. lon-1 encodes a 312 amino acid protein with a motif sequence that is conserved from plants to humans. Expression studies confirm that LON-1 is repressed by DBL-1, suggesting that LON-1 is a novel downstream component of the C.elegans TGF-beta growth regulation pathway. Consistent with this, LON-1 is expressed mainly in the larval and adult hypodermis and has dose-dependent effects on body length associated with changes in hypodermal ploidy, but not hypodermal cell proliferation. PMID- 11867536 TI - The roX genes encode redundant male-specific lethal transcripts required for targeting of the MSL complex. AB - The roX1 and roX2 genes of Drosophila produce male-specific non-coding RNAs that co-localize with the Male-Specific Lethal (MSL) protein complex. This complex mediates up-regulation of the male X chromosome by increasing histone H4 acetylation, thus contributing to the equalization of X-linked gene expression between the sexes. Both roX genes overlap two of approximately 35 chromatin entry sites, DNA sequences proposed to act in cis to direct the MSL complex to the X chromosome. Although dosage compensation is essential in males, an intact roX1 gene is not required by either sex. We have generated flies lacking roX2 and find that this gene is also non-essential. However, simultaneous removal of both roX RNAs causes a striking male-specific reduction in viability accompanied by relocation of the MSL proteins and acetylated histone H4 from the X chromosome to autosomal sites and heterochromatin. Males can be rescued by roX cDNAs from autosomal transgenes, demonstrating the genetic separation of the chromatin entry and RNA-encoding functions. Therefore, the roX1 and roX2 genes produce redundant, male-specific lethal transcripts required for targeting the MSL complex. PMID- 11867537 TI - Limited demethylation leaves mosaic-type methylation states in cloned bovine pre implantation embryos. AB - Cloning by nuclear transfer (NT) has been riddled with difficulties: most clones die before birth and survivors frequently display growth abnormalities. The cross species similarity in abnormalities observed in cloned fetuses/animals leads us to suspect the fidelity of epigenetic reprogramming of the donor genome. Here, we found that single-copy sequences, unlike satellite sequences, are demethylated in pre-implantation NT embryos. The differential demethylation pattern between genomic sequences was confirmed by analyzing single blastocysts. It suggests selective demethylation of other developmentally important genes in NT embryos. We also observed a reverse relationship between methylation levels and inner cell mass versus trophectoderm (ICM/TE) ratios, which was found to be a result of another type of differential demethylation occurring in NT blastocysts where unequal methylation was maintained between ICM and TE regions. TE-localized methylation aberrancy suggests a widespread gene dysregulation in an extra embryonic region, thereby resulting in placental dysfunction familiar to cloned fetuses/animals. These differential demethylations among genomic sequences and between differently allocated cells produce varied overall, but specified, methylation patterns, demonstrating that epigenetic reprogramming occurs in a limited fashion in NT embryos. PMID- 11867538 TI - Hyperacetylation of chromatin at the ADH2 promoter allows Adr1 to bind in repressed conditions. AB - We report that in vivo increased acetylation of the repressed Saccharomyces cerevisiae ADH2 promoter chromatin, as obtained by disrupting the genes for the two deacetylases HDA1 and RPD3, destabilizes the structure of the TATA box containing nucleosome. This acetylation-dependent chromatin remodeling is not sufficient to allow the binding of the TATA box-binding protein, but facilitates the recruitment of the transcriptional activator Adr1 and induces faster kinetics of mRNA accumulation when the cells are shifted to derepressing conditions. PMID- 11867541 TI - RNA quality control: degradation of defective transfer RNA. AB - The distinction between stable (tRNA and rRNA) and unstable (mRNA) RNA has been considered an important feature of bacterial RNA metabolism. One factor thought to contribute to the difference between these RNA populations is polyadenylation, which promotes degradation of unstable RNA. However, the recent discovery that polyadenylation also occurs on stable RNA led us to examine whether poly(A) might serve as a signal for eliminating defective stable RNAs, and thus play a role in RNA quality control. Here we show that a readily denaturable, mutant tRNA(Trp) does not accumulate to normal levels in Escherichia coli because its precursor is rapidly degraded. Degradation is largely dependent on polyadenylation of the precursor by poly(A) polymerase and on its removal by polynucleotide phosphorylase. Thus, in the absence of these two enzymes large amounts of tRNA(Trp) precursor accumulate. We propose that defective stable RNA precursors that are poorly converted to their mature forms may be polyadenylated and subsequently degraded. These data indicate that quality control of stable RNA metabolism in many ways resembles normal turnover of unstable RNA. PMID- 11867539 TI - Control of cytomegalovirus lytic gene expression by histone acetylation. AB - Permissiveness for human cytomegalovirus (HCMV) infection is dependent on the state of cellular differentiation and has been linked to repression of the viral major immediate early promoter (MIEP). We have used conditionally permissive cells to analyze differential regulation of the MIEP and possible mechanisms involved in latency. Our data suggest that histone deacetylases (HDACs) are involved in repression of the MIEP in non-permissive cells as inhibition of HDACs induces viral permissiveness and increases MIEP activity. Non-permissive cells contain the class I HDAC, HDAC3; super-expression of HDAC3 in normally permissive cells reduces infection and MIEP activity. We further show that the MIEP associates with acetylated histones in permissive cells, and that in peripheral blood monocytes the MIEP associates with heterochromatin protein 1 (HP1), a chromosomal protein implicated in gene silencing. As monocytes are believed to be a site of viral latency in HCMV carriers and reactivated virus is only observed upon differentiation into macrophages, we propose that chromatin remodeling of the MIEP following cellular differentiation could potentially play a role in reactivation of latent HCMV. PMID- 11867540 TI - Central role of Drosophila SU(VAR)3-9 in histone H3-K9 methylation and heterochromatic gene silencing. AB - Su(var)3-9 is a dominant modifier of heterochromatin-induced gene silencing. Like its mammalian and Schizosaccharomyces pombe homologues, Su(var) 3-9 encodes a histone methyltransferase (HMTase), which selectively methylates histone H3 at lysine 9 (H3-K9). In Su(var)3-9 null mutants, H3-K9 methylation at chromocentre heterochromatin is strongly reduced, indicating that SU(VAR)3-9 is the major heterochromatin-specific HMTase in Drosophila. SU (VAR)3-9 interacts with the heterochromatin-associated HP1 protein and with another silencing factor, SU(VAR)3-7. Notably, SU(VAR)3-9-HP1 interaction is interdependent and governs distinct localization patterns of both proteins. In Su(var)3-9 null mutants, concentration of HP1 at the chromocentre is nearly lost without affecting HP1 accumulation at the fourth chromosome. By contrast, in HP1 null mutants SU(VAR)3 9 is no longer restricted at heterochromatin but broadly dispersed across the chromosomes. Despite this interdependence, Su(var)3-9 dominates the PEV modifier effects of HP1 and Su(var)3-7 and is also epistatic to the Y chromosome effect on PEV. Finally, the human SUV39H1 gene is able to partially rescue Su(var)3-9 silencing defects. Together, these data indicate a central role for the SU(VAR)3 9 HMTase in heterochromatin-induced gene silencing in Drosophila. PMID- 11867543 TI - Protein 61K, encoded by a gene (PRPF31) linked to autosomal dominant retinitis pigmentosa, is required for U4/U6*U5 tri-snRNP formation and pre-mRNA splicing. AB - In each round of nuclear pre-mRNA splicing, the U4/U6*U5 tri-snRNP must be assembled from U4/U6 and U5 snRNPs, a reaction that is at present poorly understood. We have characterized a 61 kDa protein (61K) found in human U4/U6*U5 tri-snRNPs, which is homologous to yeast Prp31p, and show that it is required for this step. Immunodepletion of protein 61K from HeLa nuclear extracts inhibits tri snRNP formation and subsequent spliceosome assembly and pre-mRNA splicing. Significantly, complementation with recombinant 61K protein restores each of these steps. Protein 61K is operationally defined as U4/U6 snRNP-specific as it remains bound to this particle at salt concentrations where the tri-snRNP dissociates. However, as shown by two-hybrid analysis and biochemical assays, protein 61K also interacts specifically with the U5 snRNP-associated 102K protein, indicating that it physically tethers U4/U6 to the U5 snRNP to yield the tri-snRNP. Interestingly, protein 61K is encoded by a gene (PRPF31) that has been shown to be linked to autosomal dominant retinitis pigmentosa. Thus, our studies suggest that disruptions in tri-snRNP formation and function resulting from mutations in the 61K protein may contribute to the manifestation of this disease. PMID- 11867542 TI - MRM2 encodes a novel yeast mitochondrial 21S rRNA methyltransferase. AB - Mitochondria of the yeast Saccharomyces cerevisiae assemble their ribosomes from ribosomal proteins, encoded by the nuclear genome (with one exception), and rRNAs of 15S and 21S, encoded by the mitochondrial genome. Unlike cytoplasmic rRNA, which is highly modified, mitochondrial rRNA contains only three modified nucleotides: a pseudouridine (Psi(2918)) and two 2'-O-methylated riboses (Gm(2270) and Um(2791)) located at the peptidyl transferase centre of 21S rRNA. We demonstrate here that the yeast nuclear genome encodes a mitochondrial protein, named Mrm2, which is required for methylating U(2791) of 21S rRNA, both in vivo and in vitro. Deletion of the MRM2 gene causes thermosensitive respiration and leads to rapid loss of mitochondrial DNA. We propose that Mrm2p belongs to a new class of three eukaryotic RNA-modifying enzymes and is the orthologue of FtsJ/RrmJ, which methylates a nucleotide of the peptidyl transferase centre of Escherichia coli 23S rRNA that is homologous to U(2791) of 21S rRNA. Our data suggest that this universally conserved modified nucleotide plays an important function in vivo, possibly by inducing conformational rearrangement of the peptidyl transferase centre. PMID- 11867545 TI - The hepatitis C viral NS3 protein is a processive DNA helicase with cofactor enhanced RNA unwinding. AB - The RNA helicase/protease NS3 plays a central role in the RNA replication of hepatitis C virus (HCV), a cytoplasmic RNA virus that represents a major worldwide health problem. NS3 is, therefore, an important drug target in the effort to combat HCV. Most work has focused on the protease, rather than the helicase, activities of the enzyme. In order to further characterize NS3 helicase activity, we evaluated individual stages of duplex unwinding by NS3 alone and in complex with cofactor NS4A. Despite a putative replicative role in RNA unwinding, we found that NS3 alone is a surprisingly poor helicase on RNA, but that RNA activity is promoted by cofactor NS4A. In contrast, NS3 alone is a highly processive helicase on DNA. Phylogenetic analysis suggests that this robust DNA helicase activity is not vestigial and may have specifically evolved in HCV. Given that HCV has no replicative DNA intermediate, these findings suggest that NS3 may have the capacity to affect host DNA. PMID- 11867544 TI - The Khd1 protein, which has three KH RNA-binding motifs, is required for proper localization of ASH1 mRNA in yeast. AB - RNA localization is a widespread mechanism for achieving localized protein synthesis. In Saccharomyces cerevisiae, Ash1 is a specific repressor of transcription that localizes asymmetrically to the daughter cell nucleus through the localization of ASH1 mRNA to the distal tip of the daughter cell. This localization depends on the actin cytoskeleton and five She proteins, one of which is a type V myosin motor, Myo4. We show here that a novel RNA-binding protein, Khd1 (KH-domain protein 1), is required for efficient localization of ASH1 mRNA to the distal tip of the daughter cell. Visualization of ASH1 mRNA in vivo using GFP-tagged RNA demonstrated that Khd1 associates with the N element, a cis-acting localization sequence within the ASH1 mRNA. Co-immunoprecipitation studies also indicated that Khd1 associates with ASH1 mRNA through the N element. A khd1Delta mutation exacerbates the phenotype of a weak myo4 mutation, whereas overexpression of KHD1 decreases the concentration of Ash1 protein and restores HO expression to she mutants. These results suggest that Khd1 may function in the linkage between ASH1 mRNA localization and its translation. PMID- 11867546 TI - The mRNA export factor Dbp5 is associated with Balbiani ring mRNP from gene to cytoplasm. AB - The DEAD box RNA helicase Dbp5 is essential for nucleocytoplasmic transport of mRNA-protein (mRNP) complexes. Dbp5 is present mainly in the cytoplasm and is enriched at the cytoplasmic side of nuclear pore complexes (NPCs), suggesting that it acts in the late part of mRNP export. Here, we visualize the assembly and transport of a specific mRNP particle, the Balbiani ring mRNP in the dipteran Chironomus tentans, and show that a Dbp5 homologue in C.tentans, Ct-Dbp5, binds to pre-mRNP co-transcriptionally and accompanies the mRNP to and through the nuclear pores and into the cytoplasm. We also demonstrate that Ct-Dbp5 accumulates in the nucleus and partly disappears from the NPC when nuclear export of mRNA is inhibited. The fact that Ct-Dbp5 is present along the exiting mRNP fibril extending from the nuclear pore into the cytoplasm supports the view that Ct-Dbp5 is involved in restructuring the mRNP prior to translation. Finally, the addition of the export factor Dbp5 to the growing transcript highlights the importance of the co-transcriptional loading process in determining the fate of mRNA. PMID- 11867547 TI - Sequence-specific interaction of U1 snRNA with the SMN complex. AB - The survival of motor neurons (SMN) protein complex functions in the biogenesis of spliceosomal small nuclear ribonucleoprotein particles (snRNPs) and prob ably other RNPs. All spliceosomal snRNPs have a common core of seven Sm proteins. To mediate the assembly of snRNPs, the SMN complex must be able to bring together Sm proteins with U snRNAs. We showed previously that SMN and other components of the SMN complex interact directly with several Sm proteins. Here, we show that the SMN complex also interacts specifically with U1 snRNA. The stem--loop 1 domain of U1 (SL1) is necessary and sufficient for SMN complex binding in vivo and in vitro. Substitution of three nucleotides in the SL1 loop (SL1A3) abolishes SMN interaction, and the corresponding U1 snRNA (U1A3) is impaired in U1 snRNP biogenesis. Microinjection of excess SL1 but not SL1A3 into Xenopus oocytes inhibits SMN complex binding to U1 snRNA and U1 snRNP assembly. These findings indicate that SMN complex interaction with SL1 is sequence-specific and critical for U1 snRNP biogenesis, further supporting the direct role of the SMN complex in RNP biogenesis. PMID- 11867548 TI - The structure of the Dead ringer-DNA complex reveals how AT-rich interaction domains (ARIDs) recognize DNA. AB - The AT-rich interaction domain (ARID) is a DNA-binding module found in many eukaryotic transcription factors. Using NMR spectroscopy, we have determined the first ever three-dimensional structure of an ARID--DNA complex (mol. wt 25.7 kDa) formed by Dead ringer from Drosophila melanogaster. ARIDs recognize DNA through a novel mechanism involving major groove immobilization of a large loop that connects the helices of a non-canonical helix-turn-helix motif, and through a concomitant structural rearrangement that produces stabilizing contacts from a beta-hairpin. Dead ringer's preference for AT-rich DNA originates from three positions within the ARID fold that form energetically significant contacts to an adenine-thymine base step. Amino acids that dictate binding specificity are not highly conserved, suggesting that ARIDs will bind to a range of nucleotide sequences. Extended ARIDs, found in several sequence-specific transcription factors, are distinguished by the presence of a C-terminal helix that may increase their intrinsic affinity for DNA. The prevalence of serine amino acids at all specificity determining positions suggests that ARIDs within SWI/SNF related complexes will interact with DNA non-sequence specifically. PMID- 11867549 TI - Structural basis for cooperative DNA binding by two dimers of the multidrug binding protein QacR. AB - The Staphylococcus aureus multidrug-binding protein QacR represses transcription of the qacA multidrug transporter gene and is induced by multiple structurally dissimilar drugs. QacR is a member of the TetR/CamR family of transcriptional regulators, which share highly homologous N-terminal DNA-binding domains connected to seemingly non-homologous ligand-binding domains. Unlike other TetR members, which bind approximately 15 bp operators, QacR recognizes an unusually long 28 bp operator, IR1, which it appears to bind cooperatively. To elucidate the DNA-binding mechanism of QacR, we determined the 2.90 A resolution crystal structure of a QacR-IR1 complex. Strikingly, our data reveal that the DNA recognition mode of QacR is distinct from TetR and involves the binding of a pair of QacR dimers. In this unique binding mode, recognition at each IR1 half-site is mediated by a complement of DNA contacts made by two helix-turn-helix motifs. The inferred cooperativity does not arise from cross-dimer protein-protein contacts, but from the global undertwisting and major groove widening elicited by the binding of two QacR dimers. PMID- 11867550 TI - Functional proteomics of transforming growth factor-beta1-stimulated Mv1Lu epithelial cells: Rad51 as a target of TGFbeta1-dependent regulation of DNA repair. AB - Transforming growth factor-beta (TGFbeta) conveys regulatory signals through multiple intracellular pathways, subsequently affecting various cellular functions. To identify new targets for TGFbeta, we studied the changes in the proteome of Mv1Lu lung epithelial cells in response to TGFbeta1 treatment. Thirty eight non-abundant protein spots, affected by TGFbeta1, were selected, and proteins were identified by peptide mass-fingerprinting (PMF). Among them, proteins involved in regulation of immune response, apoptosis, regulation of TGFbeta signalling, metabolism and DNA repair were identified. Twenty-eight of the 38 proteins are new targets for TGFbeta1, thus suggesting novel ways of integration of TGFbeta signalling in intracellular regulatory processes. We show that TGFbeta1-dependent decrease in expression of one of the new targets, Rad51, correlates with a decrease in DNA repair efficiency. This was evaluated by formation of nuclear Rad51-containing DNA repair complexes in response to DNA damage, by single cell gel electrophoresis and by cell survival assay. The TGFbeta1-dependent inhibition of DNA repair was reversed by ectopic overexpression of Rad51. Therefore, TGFbeta can promote DNA instability through down-regulation of Rad51 and inhibition of DNA repair. PMID- 11867560 TI - Modeling the presentation of C3d-coated antigen by B lymphocytes: enhancement by CR1/2-BCR co-ligation is selective for the co-ligating antigen. AB - We have used a set of single-chain variable fragment antibodies (sc) genetically fused with an influenza hemagglutinin-derived peptide as a means to investigate the role of CR1 and CR2 in antigen presentation by B cells. When incubated with the B cell lymphoma 2PK3, peptide-containing sc specific for either CR1 or CR1/2 mediated activation of the hemagglutinin peptide-specific T cell line IP-12-7, as assessed by IL-2 production. Efficient presentation was dependent on the binding of the constructs to CR1/2, implying that receptor-mediated endocytosis is responsible for the effect. Cross-linkage of CR1/2 or CD19 by mAb did not increase the extent of T cell activation. However, when CR1/2 was co-ligated with the BCR--using either polyclonal goat anti-mouse IgG or recombinant protein LA- the antigen concentration required to activate T cells decreased by two orders of magnitude. Moreover, this enhancement was selective for the antigen included in these complexes and did not affect the presentation of a free peptide or of antigen bound to CR1/2 excluded from the complexes. These results suggest that B cells may bind various C3d-coated antigens at a time, but only the one which reacts with the BCR will be processed with high efficiency. This mechanism may ensure the specificity of cognate T cell help. PMID- 11867561 TI - CD53, a thymocyte selection marker whose induction requires a lower affinity TCR MHC interaction than CD69, but is up-regulated with slower kinetics. AB - The molecular mechanisms that govern the survival, maturation and export of thymocytes are the subject of intense study, and candidates for involvement in these processes might be identified by their differential expression during thymocyte selection. One such molecule is the tetraspanin CD53, which is not expressed on most CD4(+)CD8(+) double-positive (DP) cells in the normal mouse. We have examined CD53 expression on DP from several class I- and class II-restricted TCR transgenic (Tg) mice, and have found a strong correlation between CD53 expression and positive selection. CD53 expression in DP was formally demonstrated to be dependent upon MHC recognition as evidenced by studying DP from MHC-deficient mice which totally lack expression of this molecule. This link between selection and CD53 expression was reminiscent of CD69, and indeed the majority of selected DP from normal mice that express CD53 also express CD69. We compared CD53 and CD69 induction in vitro using pre-selected thymocytes from TCR Tg mice that were stimulated either with mAb against TCR or with antigen presenting cells (APC) pulsed with peptides. The data shows that with either stimulus, CD69 is induced rapidly on the thymocyte surface with expression detected in as little as 2 h. CD53 induction is slower with maximal expression taking up to 20 h. We also stimulated pre-selected thymocytes from the OT-1 TCR Tg strain with APC pulsed with peptides of varying affinities for the TCR. Here low-affinity peptides which induce CD69 expression poorly were able to induce significant levels of CD53 expression. These data demonstrate that the induction of CD53 and CD69 upon selection is not identical. Thus a combination of the CD69 and CD53 selection markers may be a powerful tool to isolate thymocytes that have either been very recently selected or have arisen from differing MHC--TCR affinity interactions during selection. PMID- 11867562 TI - Reversible CD8 expression induced by common cytokine receptor gamma chain dependent cytokines in a cloned CD4(+) T(h)1 cell line. AB - T cells that are intrathymically lineage committed are believed to maintain their CD4 or CD8 co-receptor expression. Here, we investigated whether intrathymic lineage commitment involves irreversible genetic modification or whether co receptor expression can be reprogrammed depending on external stimuli. The CD4(+) T(h)1 clone 2D6 established from splenic T cells as an IL-12-dependent line survived in culture with IL-2, IL-7 or IL-15 alone. Surprisingly, CD8 expression occurred in 2D6 cells upon replacement of IL-12 with any one of the three cytokines that stimulate the common cytokine receptor gamma chain, yielding CD4(+)CD8(+) 2D6 cells. CD8 expression declined when IL-2 was replaced with IL-12 and CD8 induction was inhibited when IL-12 was included in IL-2 or IL-7 culture. Our observations show that even a lineage-committed mature T cell can be reprogrammed for co-receptor expression in response to particular external stimuli. PMID- 11867551 TI - A genetic screen for improved plasmid segregation reveals a role for Rep20 in the interaction of Plasmodium falciparum chromosomes. AB - Bacterial plasmids introduced into the human malaria parasite Plasmodium falciparum replicate well but are poorly segregated during mitosis. In this paper, we screened a random P.falciparum genomic library in order to identify sequences that overcome this segregation defect. Using this approach, we selected for parasites that harbor a unique 21 bp repeat sequence known as Rep20. Rep20 is one of six different repeats found in the subtelomeric regions of all P.falciparum chromosomes but which is not found in other eukaryotes or in other plasmodia. Using a number of approaches, we demonstrate that Rep20 sequences lead to dramatically improved episomal maintenance by promoting plasmid segregation between daughter merozoites. We show that Rep20(+), but not Rep20(-), plasmids co localize with terminal chromosomal clusters, indicating that Rep20 mediates plasmid tethering to chromosomes, a mechanism that explains the improved segregation phenotype. This study implicates a direct role for Rep20 in the physical association of chromosome ends, which is a process that facilitates the generation of diversity in the terminally located P.falciparum virulence genes. PMID- 11867563 TI - Antioxidants inhibit mercuric chloride-induced early vasculitis. AB - In the Brown Norway (BN) rat, mercuric chloride (HgCl(2)) induces a T(h)2 dominated autoimmune syndrome which includes an early phase of mast cell dependent vasculitis. We have shown in vitro that oxidative stress up-regulates IL-4 in mast cells and predisposes to degranulation. The aim of this study was to determine whether administration of antioxidants inhibits HgCl(2)-induced early vasculitis in vivo, and, if so, to examine whether modulation of the oxidative/antioxidative balance influences IgE and IL-4 expression by mast cells in situ. Groups of rats were given HgCl(2) + saline, HgCl(2) + N-acetyl-L cysteine (NAC), saline + saline or saline + NAC respectively and blood was taken and animals killed 48 h later. NAC significantly reduced both HgCl2-induced early vasculitis and HgCl(2)-enhanced IgE expression on mast cells with a trend to a decrease in HgCl(2)-enhanced IL-4 expression in these cells. In addition, there was an increased rat mast cell protease (RMCP) II concentration in the serum after HgCl(2) injection and the elevated levels of RMCP II stimulated by HgCl(2) were totally abolished by the administration NAC in the HgCl(2) + NAC group. However, there was no significant change in serum total IgE concentrations between the HgCl(2) + saline group and the HgCl(2) + NAC group. The non sulphydryl-containing antioxidants desferrioxamine and pyruvate demonstrated a similar effect in inhibiting HgCl(2)-induced early vasculitis. Our data show that administration of an antioxidant to BN rats reduces HgCl(2)-induced early vasculitis, suggesting that oxidative stress plays a role in the pathogenesis of HgCl(2)-induced early vasculitis. This finding may have implications for the understanding of the initiation in this experimental model of T(h)2 cell-driven autoimmunity and possibly of analogous human diseases. PMID- 11867564 TI - Expression and function of 4-1BB and 4-1BB ligand on murine dendritic cells. AB - 4-1BB (CDw137) and its ligand (4-1BBL) have been implicated in cellular immune responses. To further characterize the expression and function of 4-1BBL, we newly generated an anti-mouse 4-1BBL mAb (TKS-1), which can inhibit the interaction of 4-1BBL with 4-1BB. Flow cytometric analyses using TKS-1 and an anti-mouse 4-1BB mAb indicated that 4-1BB was inducible on both CD4(+) and CD8(+) splenic T cells by stimulation with immobilized anti-CD3 mAb, but 4-1BBL was not expressed on resting or activated T cells. 4-1BBL expression was inducible on splenic B cells by stimulation with anti-IgM antibody plus anti-CD40 mAb, on peritoneal macrophages by stimulation with lipopolysaccharide (LPS) and on splenic dendritic cells (DC) by stimulation with anti-CD40 mAb or LPS. Interestingly, splenic DC expressed 4-1BB constitutively, which was down regulated by anti-CD40 stimulation. Co-culture of splenic DC with 4-1BBL transfected cells or 4-1BBL-expressing tumor cell lines led to cytokine (IL-6 and IL-12) production and co-stimulatory molecule up-regulation by splenic DC, indicating that 4-1BBL can directly activate DC. Moreover, IL-12 production by anti-CD40-stimulated DC was partially inhibited by TKS-1. These results suggest that 4-1BB expressed on DC may be involved in DC activation through DC--tumor interaction and DC--DC interaction. PMID- 11867565 TI - S100A8, S100A9 and the S100A8/A9 heterodimer complex specifically bind to human endothelial cells: identification and characterization of ligands for the myeloid related proteins S100A9 and S100A8/A9 on human dermal microvascular endothelial cell line-1 cells. AB - The natural ligands of the S100 EF hand proteins S100A8 and A9 [myeloid-related proteins 8 and 14] have long been searched for in order to further the understanding of the role of the S100A8/A9-expressing monocyte subpopulation in progressing inflammatory processes. We demonstrate that S100A8, S100A9 and the S100A8/A9 heterodimeric complex bind to human dermal microvascular endothelial cell line (HMEC)-1 with an increasing binding capacity progressing from S100A8 < or = S100A9 < or = S100A8/A9. Similar results were obtained in the apolipoprotein E knockout mouse model, where preferably recombinant S100A9 but no S100A8 bound to the endothelium of the aorta ascendens. The binding of the S100A8/A9 heterodimer complex to activated HMEC-1 is specific as demonstrated by a dose responding and satiable binding curve and the competition of FITC-labeled versus unlabeled protein. The protein character of the binding site was proven by treatment with trypsin. S100A8/A9 binding to HMEC-1 is inducible by lipopolysaccharide and tumor necrosis factor-alpha, and in the presence of calcium. A 163-kDa protein was isolated from a cell lysate of activated HMEC-1 cells using an affinity-chromatography protocol. The endothelial cell-associated ligand proteins isolated by the use of the S100A9 monomer and the S100A8/A9 dimer were subjected to mass spectrometry for protein identification. Clearly, alpha(2) macroglobulin was identified as a binding partner for the S100A9 monomer, whereas no protein could be identified from the database for the ligand of the S100A8/A9 dimer. PMID- 11867566 TI - The Trypanosoma cruzi trans-sialidase is a T cell-independent B cell mitogen and an inducer of non-specific Ig secretion. AB - Polyclonal lymphocyte activation and hypergammaglobulinemia characterize the acute phase of many parasitic diseases, including Chagas' disease, a debilitating condition caused by Trypanosoma cruzi. Polyclonal lymphocyte activation correlates with disease susceptibility inT. cruzi infection. Thus, identifying factors that drive such reactivities should provide insight into mechanisms of parasite evasion from host immunity and of disease pathogenesis. Sensitization of mice with small doses of T. cruzi trans-sialidase (TS) turns the mice into highly susceptible hosts to T. cruzi. In addition, TS heterologously expressed in Leishmania major greatly enhances virulence of the parasite to mice. In attempt to study the mechanism of TS-induced virulence, we found that TS and its C terminal long tandem repeat (LTR) are T-independent polyclonal activators for mouse B cells. While B cells deficient/defective in L-6, CD40 or Toll-like receptor-4 are similarly activated by TS as compared to wild-type cells, B cells from Bruton's tyrosine kinase-defective X-linked immunodeficient mice are remarkably insensitive to TS activation. TS-induced B cell activation in vitro is accompanied by Ig secretion independent of T cells. Furthermore, administration of TS into normal mice leads to non-specific Ig secretion that peaks 4-6 days after injection. Thus TS, through its LTR, induces abnormal polyclonal B cell activation and Ig secretion, which could explain in part its virulence-enhancing activity. PMID- 11867567 TI - Overexpression of rab7 enhances the kinetics of antigen processing and presentation with MHC class II molecules in B cells. AB - rab7 is an intracellular GTPase involved in early to late endosome fusion. By overexpressing rab7 in a B lymphoma we show that the rate of antigen presentation with MHC class II molecules is increased for four different peptide-MHC combinations, under conditions where levels of other components of the antigen processing pathway remained constant. Resting B cells were shown to express significantly lower levels of rab7 when compared to adherent macrophages or to 'immature' or 'mature' dendritic cells. rab7 expression was up-regulated by stimulation of B cells with lipopolysaccharide or CD40 ligand. Other components of the endocytic pathway were also up-regulated in activated B cells, suggesting that B cell activation leads to a general enlargement of the endocytic compartment, correlating with the increased ability of activated B cells to process antigen. Taken together, our results suggest that rab7 levels regulate the rate of antigen presentation in B cells, and that rab7 and late endocytic compartments are important in MHC class II-restricted antigen presentation in B cells. PMID- 11867568 TI - Dissection of B cell differentiation during primary immune responses in mice with altered CD40 signals. AB - CD40 is essential for efficient humoral immune responses. CD40 has two cytoplasmic domains required for binding of tumor necrosis factor receptor associated factors (TRAF). The TRAF6-binding site is within the membrane proximal cytoplasmic (Cmp) region, while a PXQXT motif in the membrane distal cytoplasmic (Cmd) region needs to engage TRAF2/3/5. To dissect CD40 signals necessary for B cell differentiation, we generated transgenic mice expressing wild-type and mutant human CD40 (hCD40) molecules in a mouse CD40-deficient (mCD40(-/-)) background. The B cell-specific expression of hCD40 in mCD40(-/-) mice resulted in T-dependent antibody responses including germinal center (GC) formation. Mutant hCD40 molecules that carry either a point mutation of the TRAF2/3/5 binding site or a deletion of the Cmd region rescued extrafollicular B cell differentiation but not GC formation. A mutant hCD40 that comprises of only the TRAF2/3/5-binding site in the cytoplasmic region also rescued low but significant titers of antigen-specific IgG1 without GC formation. These results demonstrated that two distinct signals either from the Cmp or from the Cmd region induced the extrafollicular B cell differentiation and Ig class switching; however, GC formation required both. We conclude that combinations of these two signals determine which of the extrafollicular or the follicular (GC) differentiation pathway B cells enter. PMID- 11867569 TI - Functional demonstration of intrathymic binding sites and microvascular gates for prothymocytes in irradiated mice. AB - Quantitative intrathymic (i.t.) and i.v. adoptive transfer assays for prothymocytes show strict log dose saturation kinetics, consistent with a finite number of i.t. binding sites (microenvironmental niches). This inference is supported here by demonstration of competitive antagonism obeying one-on-one receptor occupancy kinetics during the establishment of thymic chimerism in irradiated adult mice. The results of primary and secondary transfer experiments suggested that hematogenous precursors (i) enter specific i.t. niches between 4 and 24 h after injection, (ii) compete reversibly with subsequently introduced precursors, (iii) establish insurmountable competition within 5-7 days, (iv) mature through the initial stages of thymocytopoiesis preceding proliferative expansion, and (v) vacate the niches between 7 and 14 days after entry. The results also suggested that, as in non-irradiated mice, prothymocyte importation in irradiated mice is a gated phenomenon. Gate closure was indicated by the inability of i.v.-, but not i.t.-, injected bone marrow (BM) cells to induce thymic chimerism when administered 7--14 days after a primary injection and gate opening by the ability of i.v.-injected BM cells to induce thymic chimerism in competition with circulating host prothymocytes. Gate closing was log dose responsive and could be induced in individual thymic lobes by unilateral i.t. injection, whereas gate opening, which occurs bilaterally, was not initiated until most of the niches for prothymocytes had been vacated. We therefore posit the existence of a series of associated microvascular gates and microenvironmental niches that act in concert to regulate prothymocyte importation and early thymocyte differentiation. PMID- 11867570 TI - Characterization of nitrergic neurons in the porcine and human ciliary nerves. AB - PURPOSE: To further characterize a subpopulation of choroidal ganglion cells associated with the ciliary nerves. METHODS: Isolated long ciliary nerves of porcine and human eyes containing ciliary nerve-associated ganglion cells (CNGCs) were embedded in Epon for ultrastructural investigation, or wholemounts were stained with antibodies against nitric oxide synthase (NOS), vasoactive intestinal polypeptide (VIP), vesicular acetylcholine transporter, neuropeptide Y (NPY), tyrosine hydroxylase (TH), calcitonin gene-related peptide (CGRP), substance P (SP), and synaptophysin. In addition, wholemount preparations of the choroid and of the anterior segment were stained for reduced nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-D). Serial sections through choroid and anterior segment were stained with the prior antibodies listed. RESULTS: In the porcine choroid only CNGCs were present. They stained for brain (b)NOS and VIP and were surrounded by SP and VIP-immunoreactive (IR) nerve terminals. The axonal processes of the CNGCs followed the ciliary nerves to the anterior eye segment, where they formed a nerve fiber plexus that terminated in the trabecular meshwork. None of the axons passed into the sparse NOS-IR nerve fiber plexus surrounding the choroidal vasculature. The CNGCs in the human choroid morphologically resembled those seen in the pig. CONCLUSIONS: The CNGC proportion of choroidal ganglion cells is presumably involved in the intrinsic (peripheral) innervation of the aqueous outflow tissues and of the choroid. PMID- 11867571 TI - Cytoarchitecture of the retinal ganglion cells in the rat. AB - PURPOSE: To determine the number and cytoarchitecture of retinal ganglion cells (RGCs) in the female Wistar rat, by using a newly devised procedure for rapid RGC counting in the entire retina that avoids assumptions about RGC spatial arrangement. METHODS: RGCs of normal female Wistar rats were retrogradely labeled with a fluorescent tracer. Automated counting was accomplished by applying standard imaging software to analysis of all labeled cells in retinal flatmounts. The method was validated by comparison of automated and manual counts of 70,000 RGCs in frames covering the density range in the normal rat retina of 600 to 3600 RGC/mm(2). RGC numbers were determined for each retina and compared with the contralateral retina of the same animal. RGC density maps were constructed for each retina. RGC size distribution was determined. RESULTS: Automated RGC counting showed a good linear correlation with manual counting (R(2) = 0.9416). Mean total RGC count in 10 rat eyes was 97,609 +/- 3,930 (SEM) per eye. Contralateral eyes differed by an average of 4.1% (3983 plus minus 5098 RGCs). Size analysis calculated from cell areas confirmed that the majority of rat RGCs are between 7 and 21.5 microm in equivalent diameter. The RGC counts for all frames at the same eccentricity in all 10 of the retinas showed that variability increased with eccentricity and increased further as the fractional area of the retina sampled at each eccentricity was reduced. There was also significant variability in the spatial density of the RGCs at the same eccentricity location between different eyes. Comparison of total RGC counts between left and right eyes estimated from RGC counts in sectors of the retina (hemiretinas or quadrants) showed increased variability compared with counting all the RGCs in a retina. CONCLUSIONS: RGCs in the Wistar rat display significant variability in their cytoarchitecture. Such variability can make quantification by sampling problematic for diffuse, and particularly, for focal RGC losses resulting from experimental interventions, unless virtually the entire RGC population is counted. PMID- 11867572 TI - Structure and function of the N-cadherin/catenin complex in retinoblastoma. AB - PURPOSE: To identify in human retinoblastoma and normal retinal tissue the type of cadherin, its relationship with cytoplasmic catenins, and its participation in invasion. METHODS: The cadherin/catenin complex was characterized in surgical retinoblastoma specimens from five patients and human retinas from four donor eyes by immunocytochemistry, flow cytometry, and coimmunoprecipitation with antibodies against N-cadherin, alpha-catenin, and beta-catenin, followed by Western blot analysis or autoradiography. Y79 and WERI-Rb-1 retinoblastoma cell lines serve the evaluation of the cadherin/catenin complex in aggregation and collagen type I invasion in vitro. The association of the cadherin/catenin complex with the cytoskeleton was examined by an antibody-capping assay. RESULTS: In retinoblastoma and normal retina N-cadherin associated with alpha-catenin and beta-catenin but not E- or P-cadherin. The N-cadherin/catenin complex formed a regular, linear, and continuous honeycomb pattern in normal retina that was irregular, clustered, and interrupted in retinoblastoma. The N-cadherin/catenin complex was found also in the retinoblastoma cell lines WERI-Rb and Y79, in which it also showed an irregular pattern. Both cell lines were invasive in collagen type I, and invasion was inhibited by the GC-4 antibody, which functionally neutralizes N-cadherin. Less GC-4 antibody was needed to inhibit invasion of Y79 cells, which expressed N-cadherin at a lower level, than to inhibit invasion of WERI-Rb-1 cells. In both cell lines, antibody capping of the N-cadherin/catenin complex indicated that its linkage with the cytoskeleton were weak or absent. CONCLUSIONS: Retinoblastoma cells, in contrast with normal retina, express an N cadherin/catenin complex that is irregularly distributed and weakly linked to the cytoskeleton. In retinoblastoma, this complex acts as an invasion promoter. PMID- 11867573 TI - Gene expression profile of native human retinal pigment epithelium. AB - PURPOSE: To generate a profile of genes expressed in the native human retinal pigment epithelium and identify candidate genes for retinal and macular diseases. METHODS: Two cDNA libraries (one amplified, the other unamplified) were constructed using RNA isolated from native human RPE sheets. The sequence from the 5' end was obtained for randomly selected clones from the two libraries. Of these, more than 2000 expressed sequence tags (ESTs) were analyzed for similarity to sequences and gene clusters in public databases. RESULTS: EST analysis revealed several known RPE-expressed genes and more than 500 genes that have been characterized previously but were not known to be expressed in the RPE. Transthyretin and 90-kDa heat shock protein represent the most abundant transcripts identified in these RPE libraries. More than 200 novel ESTs and putative proteins were identified. An additional 344 sequences matched only the human genomic sequence. CONCLUSIONS: High-complexity cDNA libraries were generated from native human RPE. Analysis of ESTs generated from these libraries has yielded a profile of genes expressed in the native RPE. Several of the identified genes are known to play a significant role in the RPE. Novel ESTs, putative proteins, and genomic hits may represent as yet unidentified RPE expressed genes and many of these, mapping in the region of retinal disease loci, may serve as candidate genes. In addition, the nonredundant set of more than 1100 genes and ESTs described herein will be a valuable resource for generating gene microarrays, which can assist in delineating RPE expression profiles during human disease pathogenesis. PMID- 11867574 TI - Blindness, visual impairment and the problem of uncorrected refractive error in a Mexican-American population: Proyecto VER. AB - PURPOSE: To report the prevalence of blindness and visual impairment and the contribution of uncorrected refractive error to visual loss, in a population based sample of Mexican Americans aged 40 and older. METHODS: Proyecto VER is a population-based study of blindness and visual impairment in Mexican Americans in Arizona. Block groups in Tucson and Nogales were randomly selected with probability proportional to the size of the Mexican-American population aged 40 and older. Participants had a complete ophthalmic evaluation, including assessment of presenting and best corrected visual acuity using standardized procedures. Those with presenting visual acuity worse than 20/30 had refraction to determine best corrected vision. A home questionnaire and a clinic examination provided data on education, perception of visual impairment, income, and acculturation. RESULTS: The prevalence of presenting visual acuity worse than 20/40 was 8.2%, with uncorrected refractive error accounting for 73% of the impaired acuity. In multivariate models comparing those who improved two or more lines on the acuity chart with proper refraction with those who had adequate optical correction, uncorrected refractive error showed a strong association with age, less than 13 years of education (odds ratio [OR] 1.6, 95% confidence interval [CI] 1.5-2.0), low acculturation index (OR 1.3, CI 1.1-1.3), lack of insurance coverage (OR 1.4, CI 1.1-1.7), and not having seen an eye-care provider in the past 2 years (OR 2.5, CI 2.1-3.0). Prevalence of best corrected acuity worse than 20/40 increased from 0.3% in those aged 40 to 49 years to 18% in those aged 80 years or more. CONCLUSIONS: Visual loss in this Mexican-American population is higher than has been reported in whites and is comparable to that in African Americans. Almost three quarters of those with visual acuity impairment would improve with optical correction. Socioeconomic factors that are probable markers of limited access to health care services were associated with uncorrected refractive error. These data suggest that education programs and interventions to improve access to eye care could significantly decrease the burden of visual loss among Mexican Americans. PMID- 11867575 TI - Refractive error in children in a rural population in India. AB - PURPOSE: To assess the prevalence of refractive error and related visual impairment in school-aged children in the rural population of the Mahabubnagar district in the southern Indian state of Andhra Pradesh. METHODS: Random selection of village-based clusters was used to identify a sample of children 7 to 15 years of age. From April 2000 through February 2001, children in the 25 selected clusters were enumerated in a door-to-door survey and examined at a rural eye center in the district. The examination included visual acuity measurements, ocular motility evaluation, retinoscopy and autorefraction under cycloplegia, and examination of the anterior segment, media, and fundus. Myopia was defined as spherical equivalent refractive error of at least -0.50 D and hyperopia as +2.00 D or more. Children with reduced vision and a sample of those with normal vision underwent independent replicate examinations for quality assurance in seven clusters. RESULTS: A total of 4414 children from 4876 households was enumerated, and 4074 (92.3%) were examined. The prevalence of uncorrected, baseline (presenting), and best corrected visual acuity of 20/40 or worse in the better eye was 2.7%, 2.6%, and 0.78%, respectively. Refractive error was the cause in 61% of eyes with vision impairment, amblyopia in 12%, other causes in 15%, and unexplained causes in the remaining 13%. A gradual shift toward less-positive values of refractive error occurred with increasing age in both boys and girls. Myopia in one or both eyes was present in 4.1% of the children. Myopia risk was associated with female gender and having a father with a higher level of schooling. Higher risk of myopia in children of older age was of borderline statistical significance (P = 0.069). Hyperopia in at least one eye was present in 0.8% of children, with no significant predictors. CONCLUSIONS: Refractive error was the main cause of visual impairment in children aged between 7 and 15 years in rural India. There was a benefit of spectacles in 70% of those who had visual acuity of 20/40 or worse in the better eye at baseline examination. Because visual impairment can have a significant impact on a child's life in terms of education and development, it is important that effective strategies be developed to eliminate this easily treated cause of visual impairment. PMID- 11867576 TI - Refractive error in children in an urban population in New Delhi. AB - PURPOSE: To assess the prevalence of refractive error and related visual impairment in school-aged children in an urban population in New Delhi, India. METHODS: Random selection of geographically defined clusters was used to identify a sample of children 5 to 15 years of age. From December 2000 through March 2001, children in 22 selected clusters were enumerated through a door-to-door survey and examined at a local facility. The examination included visual acuity measurements, ocular motility evaluation, retinoscopy and autorefraction under cycloplegia, and examination of the anterior segment, media, and fundus. Myopia was defined as spherical equivalent refractive error of at least -0.50 D and hyperopia as +2.00 D or more. Children with reduced vision and a sample of those with normal vision underwent independent replicate examinations for quality assurance in four of the clusters. RESULTS: A total of 7008 children from 3426 households were enumerated, and 6447 (92.0%) examined. The prevalence of uncorrected, baseline (presenting), and best corrected visual acuity of 20/40 or worse in the better eye was 6.4%, 4.9%, and 0.81%, respectively. Refractive error was the cause in 81.7% of eyes with vision impairment, amblyopia in 4.4%, retinal disorders in 4.7%, other causes in 3.3%, and unexplained causes in the remaining 5.9%. There was an age-related shift in refractive error from hyperopia in young children (15.6% in 5-year-olds) toward myopia in older children (10.8% in 15-year olds). Overall, hyperopia was present in 7.7% of children and myopia in 7.4%. Hyperopia was associated with female gender. Myopia was more common in children of fathers with higher levels of education. CONCLUSIONS: Reduced vision because of uncorrected refractive error is a major public health problem in urban school aged children in India. Cost-effective strategies are needed to eliminate this easily treated cause of vision impairment. PMID- 11867578 TI - Novel characterization of MHC class II-negative population of resident corneal Langerhans cell-type dendritic cells. AB - PURPOSE: The presence of antigen-presenting cells (APCs) such as Langerhans cells (LCs), an epithelial form of dendritic cells (DCs), in corneal tissue is critical for generation of immune responses, including graft rejection and herpetic keratitis. The purpose of this study was to characterize the distribution and major histocompatibility complex (MHC) antigen expression of corneal LCs. METHODS: Normal and inflamed corneas were excised from BALB/c mice, and immunofluorescence staining for CD11c, CD11b, CD45, CD80 (B7.1), CD86 (B7.2), CD3, and MHC class II (Ia) was performed by confocal microscopy on wholemount corneal epithelium. RESULTS: CD11c(+) MHC class II-positive LCs were found in the limbus and corneal periphery, but not in the center of the normal cornea. These cells were CD45 positive, exhibiting bone marrow derivation, and CD3 and CD11b negative, confirming a DC lineage. Additionally, these cells were CD80 and CD86 negative, reflecting an immature phenotype. In the central and paracentral areas, however, significant numbers of CD11c(+) LCs were detected that expressed no MHC class II. It is important to note that although the density of the LCs declined from the limbus toward the center of the cornea, they were always present. In the inflamed cornea, the expression of MHC class II and costimulatory molecules CD80 and CD86 was significantly enhanced, and present in all parts of the cornea, in contrast to the normal cornea. CONCLUSIONS: The present study demonstrates for the first time the phenotype and distribution of MHC class II-negative LCs in the murine corneal epithelium. In the inflamed cornea, the LCs become activated as reflected by expression of B7 costimulatory markers. These changes in activation markers may provide additional information for devising novel immunomodulatory strategies. PMID- 11867577 TI - A mouse model of keratoconjunctivitis sicca. AB - PURPOSE: To evaluate the effects of pharmacologic inhibition of aqueous tear production and desiccating environmental stress on aqueous tear production, tear clearance, corneal epithelial permeability, and conjunctival epithelial morphology, proliferation, and conjunctival goblet cell differentiation. METHODS: Aqueous tear production was inhibited by applying transdermal scopolamine (scop) patches to the depilated midtail of female MC, CBA mice. Desiccating environmental stress was created by placing mice in a hood with a continuous airflow blower. Aqueous tear production and volume, tear clearance, and corneal barrier function were compared in four experimental groups: untreated control mice, mice placed in the blower hood, mice treated with a scop patch, and mice treated with a scop patch and blower hood (scop patch + blower). Cotton threads were used to evaluate aqueous tear production and volume. Tear clearance was assessed by fluorometric measurement of collected tear fluid 15 minutes after instillation of 1% sodium fluorescein. Corneal epithelial barrier function was assessed by fluorometric measurement of carboxyfluorescein uptake. Conjunctival morphology and goblet cell density were evaluated in stained histologic sections. Conjunctival epithelial proliferation was assessed by bromodeoxyuridine (BrdU) labeling. RESULTS: Significant decreases in cotton thread wetting and tear clearance were observed in mice treated with a scop patch (P < 0.001) or with a scop patch and blower desiccation (P < 0.001), with a greater reduction in tear clearance in the latter group. Significantly increased corneal carboxyfluorescein uptake was noted in the scop patch group compared with untreated mice (P = 0.05) and in the scop patch + blower group compared with all the other groups (P < 0.0001). Changes in conjunctival epithelial morphology and a significant decrease in conjunctival goblet cell density (P < 0.001) were observed in the scop patch + blower group compared with the untreated control group. The number of proliferating conjunctival epithelial cells was significantly greater in the scop patch + blower group. CONCLUSIONS: Pharmacologic inhibition of tear production in mice is accompanied by ocular surface epithelial changes resembling human keratoconjunctivitis sicca (KCS) that are exacerbated by desiccating environmental stress. Cholinergic stimulated tear production appears to be essential for maintaining a healthy ocular surface. PMID- 11867579 TI - Effect of immunosuppression on outcome measures in a model of rat limbal transplantation. AB - PURPOSE: To examine the effect of immunosuppression with intramuscularly injected cyclosporine and topical corticosteroid on limbal allograft survival in a new model in the inbred rat. METHODS: Orthotopic limbal tissue harvested from male Fischer 344 (isografts) or male Wistar-Firth donors (allografts) was sutured into superior and inferior lamellar excision sites in female recipient Fischer 344 rats. Grafts were examined clinically for at least 55 days. Superficial epithelial cells were sampled weekly, and the DNA extracted and probed for the male-specific gene Sry by polymerase chain reaction. Recipients were killed at established intervals for immunohistochemistry. Graft-recipient animals were randomly assigned to receive either intramuscular cyclosporine plus topical prednisolone phosphate or vehicle for 4 weeks from the time of transplantation. RESULTS: Isografts survived for a median of more than 55 days. Allografts underwent clinical rejection at a median of 6 to 7 days after grafting. Acutely rejecting allografts showed a dense mononuclear infiltrate consisting of activated CD4(+) and CD8(+) T cells with some macrophages. Genomic Sry was usually detectable in cells sampled from the ocular surface for more than 55 days in isografts, but not beyond 7 days in allografts. Immunosuppression prolonged allograft survival significantly, as assessed clinically, but did not prolong donor cell survival on the ocular surface, as assessed by detection of genomic Sry. CONCLUSIONS: A robust model of limbal transplantation was developed in the rat. Isografts survived for the long term, whereas allografts underwent rapid rejection. Although clinical allograft survival was prolonged to a modest extent by immunosuppression, donor cell survival on the ocular surface was not improved. PMID- 11867580 TI - Molecular properties of wild-type and mutant betaIG-H3 proteins. AB - PURPOSE: BetaIG-H3 is a TGF-beta-induced cell adhesion molecule, the mutations of which are responsible for a group of 5q31-linked corneal dystrophies. The characteristic findings in these diseases are accumulation of protein deposits of different ultrastructures. To understand the mechanisms of protein deposits in 5q31-linked corneal dystrophies, the molecular properties of betaIG-H3 and the effects of mutation on these properties were studied in vitro. METHODS: Substitution mutations were generated by two-step PCR. Wild-type and mutant recombinant betaIG-H3 proteins were raised in Escherichia coli. For structural study, nondenaturing gel electrophoresis, cross-linking experiments, and electron microscopy examination were performed. A solid-phase interaction assay was performed for the interaction of betaIG-H3 with other matrix proteins. Wild-type and mutant betaIG-H3 cDNAs were cloned into a mammalian expression vector and overexpressed in the corneal epithelial cells by transient transfection. Immunoprecipitation and immunoblot analysis were performed with an antibody against human betaIG-H3. Cell adhesion was assayed by measuring enzyme activities of N-acetyl-beta-D-glucosaminidase. RESULTS: The recombinant betaIG-H3 protein self-assembled to form multimeric bands and appeared to have a fibrillar structure. Solid-phase in vitro interaction assay showed that it bound strongly to type I collagen, fibronectin, and laminin; moderately to collagen type II and VI; and minimally to collagen type IV. Five recombinant mutant forms of betaIG-H3 (R124C, R124H, R124L, R555W, and R555Q) commonly found in 5q31-linked corneal dystrophies did not significantly affect the fibrillar structure, interactions with other extracellular matrix proteins, or adhesion activity in cultured corneal epithelial cells. In addition, the mutations apparently produced degradation products similar to those of wild-type betaIG-H3. CONCLUSIONS: BetaIG H3 polymerizes to form a fibrillar structure and strongly interacts with type I collagen, laminin, and fibronectin. Mutations found in the 5q31-linked corneal dystrophies do not significantly affect these properties. The results suggest that mutant forms of betaIG-H3 may require other cornea-specific factors, to form the abnormal accumulations in 5q31-linked corneal dystrophies. PMID- 11867581 TI - Assessment of ocular counterroll during head tilt using binocular video oculography. AB - PURPOSE: According to recent literature, the presence and the amount of true compensatory ocular counterroll is still debatable. The purpose of the current study was to assess compensatory counterroll in response to lateral head tilt using a new noninvasive recording technique, and, furthermore, to find out whether the amount of counterroll is influenced by the presence or absence of spatial orientation. METHODS: Eye movement recordings were performed using the infrared three-dimensional video oculography (3D-VOG) technique. Objective cycloposition of five healthy individuals was measured in presumed primary position and in head tilt positions of 15 degrees, 30 degrees, and 45 degrees to the right and left. The same paradigm was performed under three viewing conditions: binocularly without spatial orientation and both binocularly and monocularly with spatial orientation. RESULTS: A consistent ocular counterroll corresponding to the amount of head tilt was observed in all subjects. Maximum torsional amplitude was 10 degrees at a 45-degree head tilt. The relative amount of compensation ranged between 13% and 22% of the actual head tilt, decreasing with increasing head tilt. Compensatory counterroll and torsional conjugacy between both eyes revealed minor differences between the experimental paradigms. Incomplete cycloductional reorientation in primary position after head tilt was detected in all subjects, regardless of the stimulus. CONCLUSIONS: A consistent compensatory ocular counterroll was demonstrated in response to static lateral tilting of the head in healthy individuals. The amplitude of counterroll and the gain of compensatory cycloversion were higher than has been generally reported. Infrared 3D-VOG technique was a reliable and comfortable method for the assessment of ocular cycloduction. It can be considered to be a promising tool for advanced evaluation of disturbances of the oblique eye muscles. PMID- 11867582 TI - Changes in Listing's plane after sustained vertical fusion. AB - PURPOSE: To determine whether prolonged fusion of an imposed vertical disparity leads to a change in the orientation of Listing's plane, even when measured during monocular viewing. METHODS: Four normal subjects (age range, 24-37 years) wore Fresnel prisms of increasing power for 72 hours to produce a final left-over right disparity (range, 7-11 prism diopters [approximately 3.9 - 6.2 degrees]) that was still fusible. Eye movements were measured binocularly, using three-axis search coils, as subjects fixed on an array of light-emitting diodes (LEDs) arranged on a flat screen, 124 cm away. A regression was used to fit the data points to a plane (Listing's plane) during monocular and binocular viewing. From each planar fit, the horizontal and vertical components of primary position (the direction of gaze that is perpendicular to Listing's plane) were calculated. Baseline data were collected in the unadapted state, either just before or at least 4 days after wearing the prisms. RESULTS: After the period of viewing through the prisms, there was a change in vertical phoria (prism adaptation) ranging from 1.6 to 3.3. There was a significant (P < 0.01) shift of the relative orientation of the vertical component of primary position between the two eyes of 6.3 +/- 1.7 degrees (right eye value minus left eye, up being positive, each measured during monocular viewing). There was no consistent pattern of change in the horizontal component of primary position. CONCLUSIONS: Prolonged fusion of a vertical disparity is associated with a change in the orientation of Listing's plane that persists under monocular viewing. Possible mechanisms include phoria adaptation, the prolonged fusional effort itself, and the residual disparity that must be overcome by sensory mechanisms. PMID- 11867583 TI - Impaired vertical phoria adaptation in patients with cerebellar dysfunction. AB - PURPOSE: To determine whether phoria adaptation to a vertical prism disparity is altered in patients with cerebellar dysfunction. METHODS: With a computer-aided haploscope, adaptive responses of fusion-free eye position to a 10- or 30-minute period was measured in subjects wearing a 3-prism diopter vertical prism over one eye. Thirteen patients with well-documented cerebellar diseases who did not have manifest ocular misalignment or limited versional eye movement and age-matched healthy subjects participated. RESULTS: The mean +/- SD percentage of vertical phoria adaptation was 13% +/- 22% and 20% +/- 16% for the 10- and 30-minute adaptations, respectively. These levels were significantly smaller than the respective ones in the age-matched control group (P < 0.001, repeated measures MANOVA). Seven (54%) of 13 patients, including two with genetically confirmed pure cerebellar lesions (spinocerebellar ataxia type 6), showed markedly reduced responses to both the 10- and 30-minute adaptations. In all three patients with acute cerebellar ataxia, the adaptive response was improved at the same time as remission of cerebellum-associated neurologic deficits. CONCLUSIONS: Phoria adaptation to vertical binocular disparity is frequently impaired in patients with cerebellar dysfunction. These results bolster the hypothesis that phoria adaptation is a cerebellar-dependent response. PMID- 11867584 TI - Long-term effects of ricin-mAb 35 on extraocular muscles of rabbits: potential treatment for strabismus. AB - PURPOSE: The immunotoxin, ricin-mAb 35, composed of ricin conjugated to a monoclonal antibody against the nicotinic acetylcholine receptor of skeletal muscle, has been proposed as a potential new agent for treatment of focal muscle dystonias. It has been demonstrated that direct injection of ricin-mAb 35 into rabbit extraocular muscle (EOM) results in significant muscle loss within 1 week. In this study, the long-term myopathic effects of ricin-mAb 35 on extraocular muscle were investigated. METHODS: Rabbit superior rectus muscles were injected with ricin-mAb 35 at a dose of 0.2 microg/kg, with the contralateral superior rectus muscle serving as the control. After 56 days, 105 days, and 1 year, the superior rectus muscles were removed and prepared for light or electron microscopy. Postinjection changes in muscle fiber morphometry and ultrastructure were examined. Immunohistochemical markers were used to identify inflammatory cellular infiltrate and myosin heavy chain (MHC) isoform expression. RESULTS: Despite evidence of ongoing regeneration, treated muscles continued to show a decrease in both myofiber number and in total cross-sectional area 56 and 105 days after injection. Individual myofiber cross-sectional areas were markedly heterogeneous at 56 days. Myofiber number and muscle cross-sectional area returned to normal 1 year after injection, but pronounced heterogeneity of myofiber size remained. The most significant changes in myosin heavy chain (MHC) isoform expression occurred in the orbital layer, where, at 56 and 105 days, there were increased numbers of fast and neonatal myofibers and decreased numbers of slow myofibers. In the global layer, after both 105 days and 1 year, there was a decrease in myofibers that were positive for slow, neonatal, and developmental MHC expression. CONCLUSIONS: EOM injection with ricin-mAb 35 results in a sustained decrease in muscle mass at 105 days after injection, with subtler morphometric changes persisting even to 1 year. Changes in muscle force development as a result of ricin-mAb 35 injection are currently under investigation. This novel immunotoxin may be useful in the treatment of strabismus if these studies show sustained weakness in treated muscles. PMID- 11867585 TI - Slow target-directed eye movements in ataxia-telangiectasia. AB - PURPOSE: To analyze the slow eye movements that shift the direction of gaze in patients with ataxia-telangiectasia (A-T). METHODS: Eye and head movements were recorded with search coils in three patients with A-T during attempted gaze shifts, both with the head immobilized and free to move. RESULTS: Gaze shifts frequently included both saccadic and slow components. The slow movements were recorded after 42% of saccades and had an average peak velocity of 6.1 deg/sec and a mean amplitude of 2.0. They occurred with the head stationary and moving, could be directed centripetally or centrifugally, had velocity waveforms that were relatively linear or exponential, and always moved the eyes toward the visual target. CONCLUSIONS: The slow movements appear to differ from pursuit and vestibular eye movements and are not fully explained by the various types of abnormal eye movements that can follow saccades, such as gaze-evoked nystagmus or postsaccadic drift. Their origin is uncertain, but they could represent very slow saccades, due to aberrant inhibition of burst cell activity during the saccade. PMID- 11867586 TI - Apoptosis gene expression and death receptor signaling in mitomycin-C-treated human tenon capsule fibroblasts. AB - PURPOSE: To examine the effect of mitomycin-C on the expression of apoptosis genes in human Tenon capsule fibroblasts and to evaluate whether death receptor signaling modulates mitomycin-C cytotoxicity. METHODS: Bcl-2, Bax, Bcl-x, Fas (CD95) and tumor necrosis factor (TNF) receptor expression was determined by flow cytometry in control and mitomycin-C-treated Tenon fibroblasts. Fibroblast death was quantified using a lactate dehydrogenase release assay. The effect of Fas and TNF-receptor signaling was evaluated using Fas-specific antibodies and soluble TNF-alpha. RESULTS: Tenon fibroblasts constitutively express Bcl-2, Bax, and Bcl x in culture. Mitomycin-C (0.4 mg/mL) induced a small but consistent increase in the expression of all three proteins. Tenon fibroblasts express low levels of Fas but are resistant to the effects of Fas-receptor ligation. Mitomycin-C (0.01-1.0 mg/mL) led to a significant increase in Fas expression at all concentrations tested (P < 0.01). Pretreatment with mitomycin-C (0.4 mg/mL) rendered fibroblasts susceptible to agonistic anti-Fas monoclonal IgM antibodies (50-500 ng/mL) and led to a further 50% reduction in viable fibroblasts at 48 hours, compared with mitomycin-C alone (P < 0.05). Antibodies that block the Fas receptor did not inhibit mitomycin-C-induced apoptosis. CONCLUSIONS: Mitomycin-C alters apoptosis gene expression and primes fibroblasts to the effects of Fas receptor ligation. Factors other than the level of Fas receptor expression modulate the response to Fas receptor signaling. Determining the signals that regulate fibroblast apoptosis may help to refine therapeutic strategies for switching off the subconjunctival healing response and maintaining intraocular pressure control. PMID- 11867587 TI - Computational evaluation of the role of accommodation in pigmentary glaucoma. AB - PURPOSE: Accommodation has been proposed as the cause of the bowing of the posterior iris that occurs in eyes with pigmentary dispersion syndrome. A mathematical model of the anterior eye is needed to explore the elastohydrodynamic effects of accommodation on both the aqueous humor dynamics and the contour of the iris. METHODS: A mathematical model of the coupled aqueous humor-iris system was used to predict the effects of accommodation on the iris position and pressure distribution in the aqueous humor. RESULTS: The mathematical model predicts that accommodation produces a pressure reversal--the anterior chamber pressure being higher than the posterior chamber--and iris movement into a more concave configuration. Total time for accommodation, iris modulus, iris attachment point, and trabecular meshwork permeability all had little or no effect on the iris contour and pressure change. The amount of accommodation, however, had a dramatic effect on both the amount of iris curvature and especially the pressure reversal. For accommodation resulting in a 0.6-mm change in anterior chamber depth, the pressure in the anterior chamber was more than 1.0 mm Hg higher than that in the posterior chamber, compared with a pressure difference of less than 0.1 mm Hg for accommodation resulting in a 0.2 mm change in anterior chamber depth. CONCLUSIONS: The results confirm that accommodation produces bowing of the posterior iris and the magnitude of the bowing is a strong function of the amount of accommodation. PMID- 11867588 TI - Performance of efficient test procedures for frequency-doubling technology perimetry in normal and glaucomatous eyes. AB - PURPOSE: To validate the clinical performance of two new efficient threshold estimation procedures for frequency-doubling technology (FDT) perimetry in both visually normal individuals and patients with glaucomatous visual field loss. METHODS: Forty-one normal subjects (mean age, 48.3 +/- 11.6 years) and 50 patients with glaucomatous visual field loss (mean age, 72.7 +/- 10.0 years) were tested. Some of these participants were retested within a 3-month period. FDT perimetry was performed on a color monitor driven by a visual-stimulus-generating video board, with stimulus parameters designed to closely mimic those of the commercial FDT test. Visual field sensitivity was measured using three procedures: a modified binary search (MOBS) identical with the one used in the commercial FDT device, a rapid efficient binary search (REBS), and a procedure the uses Bayesian methods (zippy estimation of sequential testing; ZEST). The selection of optimum parameters for REBS and ZEST were based on results from previous simulations. RESULTS: Both ZEST and REBS were 40% to 50% faster than MOBS. All three methods produced similar visual field sensitivity measures, with 95% of the differences occurring between +/-2 dB for normal subjects and +/-3 dB for glaucoma patients. Test-retest performance was similar for all three procedures. CONCLUSIONS: The test time for full-threshold FDT perimetry can be approximately halved, by using either the ZEST or REBS procedure, without affecting the accuracy or reliability of the measurements. These findings in normal subjects and patients with glaucoma provide clinical confirmation of our previous investigations of these test strategies that use computer simulation. PMID- 11867589 TI - Metalloproteinase gene transcription in human ciliary muscle cells with latanoprost. AB - PURPOSE: The present study was undertaken to determine whether treatment of ciliary muscle cells with the prostaglandin (PG) analogue latanoprost acid alters transcription of mRNA for matrix metalloproteinase (MMP)-1, -2, -3, and -9. METHODS: Human ciliary smooth muscle cell cultures were grown to confluence and treated for 24 hours with medium supplemented with latanoprost acid or vehicle. Total RNA was then isolated, and the expression of mRNAs for MMP-1, -2, -3, and 9 were determined using Taqman and energy-transfer real-time PCR analyses. All results were normalized according to glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA in each sample. RESULTS: Specificity and linearity of each real-time PCR assay were confirmed by electrophoresis and serial dilution analysis of oligonucleotides containing the amplicon sequence. Addition of latanoprost acid for 24 hours increased expression of MMP-1 by 3- to 13-fold in three of five primary ciliary muscle lines. Addition of 8, 40, and 200 nM latanoprost acid for 24 hours increased MMP-1 mRNA in a dose-dependent manner. Analysis of cultures exposed to 200 nM latanoprost acid for 4, 6, 12, or 24 hours revealed an increase and then a decline of MMP-1 mRNA, with peak expression at 6 to 12 hours after initiation of treatment. Parallel assessments of RNA from ciliary muscle cultures exposed to latanoprost acid for 24 hours revealed increased MMP-1, -3, and -9 mRNAs and reduced MMP-2 mRNA, when compared with RNA from vehicle-treated cultures. CONCLUSIONS: Latanoprost acid induced a dose-dependent increase of MMP 1, -3, and -9 gene transcription in cultured human ciliary smooth muscle cells. These results are consistent with increased MMPs contributing to the increased uveoscleral outflow facility observed after topical latanoprost. PMID- 11867590 TI - Reliability of confocal choroidal laser Doppler flowmetry. AB - PURPOSE: To assess the variability in choroidal laser Doppler flowmetry (LDF). METHODS: Five measurements of choroidal blood flow were obtained on 5 days in both eyes of 10 subjects. Reliability, sensitivity, and sample size calculations were performed. Yield, the intensity of the returning light (direct current [DC]) divided by the signal amplification (gain) used during recording, was calculated (yield = DC/gain). The correlation between yield and the LDF parameters velocity, volume, and flux was analyzed. Reliability, sensitivity, and sample size calculations were repeated after correcting for this relationship. The influence of different light-scattering properties on the returning signal was assessed in a model eye. RESULTS: Yield and the LDF parameters, velocity, volume, and flux, correlated significantly in a regression model. After the influence of yield was partialized out, reliability, reproducibility, sensitivity, and statistical power improved markedly. The high reliability of yield in a given eye (87%) suggests, that, besides varying recording settings on different days, individual specific scattering properties within the eye influence LDF measurements. Comparison of model eye data with human data disclosed a low signal-to-noise ratio with decreasing yield. Correcting for yield did not affect the sensitivity to blood flow changes induced by hypercapnia or a suction cup. CONCLUSIONS: Tissue scattering properties alter the characteristics of the returning light, and recording settings affect the outcome of the analysis of the returning light during LDF measurements. Partializing out the influence of yield markedly improves the reproducibility of choroidal LDF. PMID- 11867591 TI - Corneal epithelial rejection in the rat. AB - PURPOSE: To investigate clinical and histologic changes in the epithelium during corneal graft rejection in the rat. METHODS: LEW (RT1(l)) or PVG (RT1(c)) strain corneas were transplanted to PVG strain recipients and examined by slit lamp for clinical signs of rejection. Recipients were killed, and corneal epithelial sheets were removed and examined by adenosine diphosphatase (ADPase) staining for Langerhans cells (LC) and by immunohistology for leukocytes and adhesion molecules (T cells, macrophages, granulocytes, major histocompatibility complex [MHC] class II, CD2 and CD54 intercellular adhesion molecule [ICAM]-1) at a range of time points before, during, and after rejection, depending on the cell type sought. Normal and contralateral eyes were examined for ADPase(+) and MHC class II(+) cells. RESULTS: Clinical rejection, as defined by stromal opacity, occurred between days 10 and 15 after transplantation. In 94% of allografts, a curved clinical epithelial rejection line was observed in which ADPase(+)/MHC class II(+), CD4(+), or CD8(+) T cells were identified. There were significantly more infiltrating cells of all types in epithelia of allografts than in those of isografts. The most numerous cells were CD4(+) and CD8(+) T cells, suggesting preferential migration of these cells into the epithelium from underlying layers. Expression of MHC class II and ICAM-1 was induced on epithelial cells. CONCLUSIONS: Epithelial rejection in rats is clinically similar to that in humans and occurs simultaneously with stromal infiltration. It may be mediated by T cells rather than macrophages. In isolation, its recognition in humans may be a useful indication that the patient is at high risk of endothelial rejection. PMID- 11867592 TI - Linkage of IL-6 with neutrophil chemoattractant expression in virus-induced ocular inflammation. AB - PURPOSE: Herpes simplex virus (HSV)-1 infection of the murine cornea is known to stimulate a vigorous interleukin (IL)-6 response, but whether this pleiotropic cytokine is an essential participant in corneal inflammation is unclear. This study was designed to compare the early inflammatory response in IL-6 gene deficient mice to that in wild-type hosts. METHODS: Gene knockout and wild-type mice (C57BL/6 background) were infected intracorneally with HSV-1 (strain RE) and observed through clinical examination and immunohistochemistry for the development of corneal opacity. Virus corneal titers were determined by standard plaque assay on Vero cells. Cytokine and chemokine levels in corneal lysates were measured with commercial ELISA kits. RESULTS: Corneal opacity in IL-6(-/-) mice was substantially diminished in comparison with IL-6(+/+) hosts 24 to 48 hours after intracorneal viral infection, and corneal levels of (MIP)-2 and MIP-1alpha were significantly reduced. Local administration of IL-6 at the time of infection restored corneal opacity and chemokine levels to that of wild-type hosts. Antibody neutralization of endogenous IL-6 in IL-6(+/+) animals reduced corneal opacity scores and MIP-2 levels to that of IL-6(-/-) mice. Ex vivo studies with excised corneal buttons revealed that uninfected IL-6(-/-) corneas injected with IL-6 produced MIP-2 and MIP-1alpha at levels comparable to that seen in IL-6(+/+) hosts. CONCLUSIONS: Collectively, these results suggest that IL-6 promotes corneal inflammation by acting in an autocrine-paracrine fashion to induce resident corneal cells to make MIP-2 and MIP-1alpha, which in turn recruit neutrophils to the virus infection site. PMID- 11867593 TI - Inhibitory effects of pyrrolidine dithiocarbamate on endotoxin-induced uveitis in Lewis rats. AB - PURPOSE: To determine the effect of pyrrolidine dithiocarbamate (PDTC), an antioxidant nuclear factor (NF)-kappaB inhibitor, on the ocular inflammation induced by lipopolysaccharide (LPS). METHODS: Endotoxin-induced uveitis (EIU) was produced by a footpad injection of 200 microg LPS in male Lewis rats. PDTC (200 mg/kg) was injected intraperitoneally 30 minutes before the LPS administration. The number of infiltrating cells and protein concentration in the aqueous humor (AqH) was determined from the AqH collected at 24 hours. Immunohistochemical staining with a monoclonal antibody against activated NF-kappaB was performed to evaluate the effect of PDTC on NF-kappaB activation. Interleukin (IL)-1beta, IL 6, and tumor necrosis factor (TNF)-alpha mRNA expression in the iris-ciliary body (ICB) was determined by RNase protection assay (RPA). The levels of these cytokines and nitric oxide (NO) production were also determined. RESULTS: The number of cells in the AqH was 1100 +/- 254 cells/microL in rats injected with LPS and 90 +/- 43 cells/microL in rats pretreated with PDTC (P < 0.001). The concentration of proteins was significantly lower in the AqH of rats pretreated with PDTC than in those without PDTC. The number of activated NF-kappaB-positive cells in the ICB was reduced by the PDTC treatment. The ICB at 6 hours after LPS injection exhibited increased expression of IL-1beta, IL-6, and TNF-alpha mRNAs, which was decreased after PDTC pretreatment. PDTC also significantly diminished the levels of these cytokines and nitrite-nitrate in the AqH. CONCLUSIONS: These results suggest that PDTC reduces ocular inflammation in eyes with EIU by downregulating proinflammatory cytokine expression and by inhibiting the NF kappaB-dependent signaling pathway. PMID- 11867594 TI - Inducible adeno-associated virus vector-delivered transgene expression in corneal endothelium. AB - PURPOSE: To investigate whether recombinant adeno-associated virus (rAAV) vector- mediated transgene expression is induced by inflammation in corneal endothelial cells in vivo. METHODS: The ocular anterior chamber of New Zealand White rabbits was injected with rAAV-LacZ (10(7) units of infection). Transient ocular anterior segment inflammation was induced by an intravitreal injection of lipopolysaccharide (LPS). The effect of inflammation on LacZ gene expression in corneal endothelial cells was evaluated by histochemical staining and reverse transcription-polymerase chain reaction (RT-PCR). The influence of rAAV on endothelial cell function was monitored by measuring corneal thickness. RESULTS: Inflammatory reaction peaked at 1 day after LPS treatment and, at the same time, most of the endothelial cells (91.3% plus minus 7.2%) showed prominent LacZ gene expression. The transgene expression gradually diminished to basal level (3.4% plus minus 2.1%) when the inflammation subsided at 15 days after LPS treatment. The diminished transgene expression was efficiently reactivated to a high level (86.1% plus minus 8.7%) by a second LPS injection 60 days later. Moreover, the transgene expression remained low for a long period (60 days) in the absence of LPS treatment, but was increased to high levels (87.3% plus minus 8.1%) 1 day after LPS treatment. Throughout the observation period, endothelial cell function remained intact. CONCLUSIONS: The rAAV vector can deliver genes into endothelial cells, and transgene expression is dramatically induced by inflammation. The rAAV delivered transgene is stable and does not compromise endothelial cell function. Inducible rAAV-mediated transgene expression in corneal endothelial cells is a potential strategy in the treatment and prevention of ocular diseases. PMID- 11867595 TI - Induction of ocular inflammation by T-helper lymphocytes type 2. AB - PURPOSE: Sight-damaging ocular inflammation is often mediated by T-helper (Th) lymphocytes. The population of Th cells is divided into two major subsets, designated Th1 and Th2, that differ by their cytokine production and biological activities. In the present study, the capacity of Th1 and Th2 cells to induce ocular inflammation was examined. METHODS: Ocular inflammation was induced in transgenic (Tg) mice that express hen egg lysozyme (HEL) in their lens, by adoptively transferring Th cells that transgenically express HEL-specific receptor. Th1 and Th2 populations were polarized in vitro, and their selective cytokine production was determined by conventional methods. Levels of ocular inflammation were monitored by conventional histologic methods. Infiltrating cells were collected from sections of inflamed eyes by microdissection, and their cytokine production was examined by RT-PCR. RESULTS: Th1 cells were highly immunopathogenic, producing disease in naive recipients at numbers as low as 0.12 x 10(6), whereas Th2 cells were inactive in these recipients, even at 30 x 10(6). Th2 cells, however, produced inflammation when transferred into sublethally irradiated recipients. Distinctive histopathologic changes characterized ocular inflammation induced by the two types of Th cells. Cytokine analysis of infiltrating cells in recipient mouse eyes, as well as of splenocytes of these mice demonstrated that the transferred cells retained their type specificity. Coinjecting Th2 and Th1 cells did not alleviate the ocular disease in naive recipients and even exacerbated the immunopathogenic process in irradiated recipients. CONCLUSIONS: Th2 cells are capable of inducing ocular inflammation, but only in immunodeficient mice, and are profoundly inferior to Th1 cells in their immunopathogenic capacity. PMID- 11867596 TI - P2Y receptor-mediated stimulation of Muller glial DNA synthesis. AB - PURPOSE: To determine whether activation of P2Y receptors may increase the DNA synthesis rate of cultured Muller cells and to investigate whether adenosine 5' triphosphate (ATP)-induced Muller cell proliferation is mediated by an intracellular calcium increase. METHODS: Primary cultures of Muller cells of the guinea pig were treated with test substances for 16 hours. The DNA synthesis rate was assessed by a bromodeoxyuridine immunoassay, and ATP-induced elevations of the intracellular calcium concentration were recorded by fura-2 imaging. RESULTS: ATP or uridine triphosphate (UTP) increased the DNA synthesis rate whereas alpha,beta-methylene-ATP, 2-methyl-thio-ATP, and adenosine were ineffective, indicating that the action of ATP was through P2Y receptors. The effect of ATP was dose dependent, with an EC(50) of 5.9 microM. The mitogenic effect of ATP required an elevation of the intracellular calcium and a calcium influx into Muller cells. Blockers of calcium-permeable channels (nickel ions) or of calcium dependent potassium (BK) channels (iberiotoxin, charybdotoxin) inhibited the ATP stimulated DNA synthesis. In calcium-imaging experiments, ATP-evoked intracellular calcium transients were significantly shortened in the presence of extracellular nickel ions or of iberiotoxin. A correlation was found between the duration of the ATP-evoked calcium transients and the basal proliferation rate of the cultures. CONCLUSIONS: The results indicate that the ATP-induced elevation of Muller glial DNA synthesis is dependent on an influx of calcium ions from the extracellular space and that the inhibiting effect of BK channel blockers on ATP evoked DNA synthesis is caused by an inhibition of this influx. The amount of the calcium influx seems to be directly correlated to the strength of the ATP-evoked proliferation. PMID- 11867597 TI - Induction of xCT gene expression and L-cystine transport activity by diethyl maleate at the inner blood-retinal barrier. AB - PURPOSE: In this study, the expression and regulation of the L-cystine transporter, system x(-)(c), at the inner blood-retinal barrier (inner BRB) was investigated using a conditionally immortalized rat retinal capillary endothelial cell line (TR-iBRB2) as an in vitro model. METHODS: For the uptake study, TR iBRB2 cells were cultured at 33C in the presence or absence of diethyl maleate (DEM), and the uptake rate of [(14)C]L-cystine was measured at 37C. The mRNA levels of system x(-)(c), which consists of xCT and 4F2hc, were determined by quantitative real-time RT-PCR analysis with specific primers. RESULTS: The xCT and 4F2hc mRNAs were expressed in TR-iBRB2 cells. The [(14)C]L-cystine uptake by TR-iBRB2 cells appeared to be mediated through a saturable Na(+)-independent process. The corresponding Michaelis-Menten constant was 9.18 microM. At 100 microM DEM, the xCT mRNA level and L-cystine uptake activity in TR-iBRB2 cells were enhanced in a time-dependent manner. Concomitantly, the glutathione concentration in TR-iBRB2 cells was increased. In contrast, the 4F2hc mRNA level was unchanged up to 24 hours and was induced for more than 24 hours by DEM treatment. Under both normal and DEM treatment conditions, the uptake of [(14)C]L cystine was strongly inhibited by L-glutamic acid, L-alpha-aminoadipic acid, L homocysteic acid, and L-quisqualic acid, whereas L-aspartic acid and L-arginine had no effect, which is evidence of the induction of system x(-)(c). CONCLUSIONS: System x(-)(c)-mediated L-cystine uptake appears to be present at the inner BRB. DEM induces L-cystine transport through system x(-)(c) at the inner BRB by enhanced transcription of the xCT gene. PMID- 11867599 TI - Evidence that nitric oxide is involved in autoregulation in optic nerve head of rabbits. AB - PURPOSE: To determine the role played by nitric oxide (NO) in the autoregulation of circulation in the optic nerve head (ONH). METHODS: The intraocular pressure (IOP) was increased and maintained at 50 mm Hg by the infusion of balanced saline solution (BSS) into the anterior chamber of albino rabbits. Experiments were performed with or without an intravenous injection of 10, 20, or 50 mg/kg N(G) nitro-L-arginine methyl ester (L-NAME), a nitric oxide synthase (NOS) inhibitor. The blood flow in the ONH was evaluated by the hydrogen clearance method, and NO metabolites (nitrite and nitrate) were measured in the ONH under the same experimental conditions in other rabbits. Visual evoked potentials (VEPs) were recorded before the IOP elevation and every 15 minutes during the 60 minutes of elevation. The effect of elevated IOP on the VEPs and the hemodynamics and NO levels in the ONH were determined. The effect of pretreatment with a NOS inhibitor on the IOP-induced changes was also investigated. RESULTS: The implicit time of the VEP was prolonged after L-NAME in a dose-dependent manner, whereas the implicit time in the control group (saline) was less affected. Blood flow in the ONH was not reduced by an elevation of the IOP (50 mm Hg) but was significantly reduced by L-NAME (20 mg/kg). The NO metabolites, which were elevated in the ONH during IOP elevation in the control, were also depressed by L NAME pretreatment. CONCLUSIONS: These results indicate that NO may play a role in the autoregulation of circulation in the ONH during elevated IOP. This would mean that NO provides some neuroprotection during an acute phase of ischemia in the ONH. PMID- 11867600 TI - Studies on endothelin release and Na,K transport in porcine lens. AB - PURPOSE: In an earlier study it was reported that thrombin significantly reduces the rate of Na,K-adenosine triphosphatase (ATPase)-mediated ion transport by porcine lens. Because thrombin stimulates the release of endogenous endothelin (ET)-1 stores from some tissues, and because ET-1 can cause Na,K-ATPase inhibition, this study was designed to determine whether thrombin causes release of ET-1 from the lens. METHODS: Intact porcine lenses were incubated in Krebs solution. The concentration of ET-1 in the solution was determined by ELISA. The rate of Na,K-ATPase-dependent ion transport was determined by measurement of ouabain-sensitive (86)Rb uptake. RESULTS: Thrombin (1 U/mL) reduced the rate of ouabain-sensitive (86)Rb uptake by approximately 40%. PD145065 (2 microM), an ET receptor antagonist, abolished the inhibitory effect of thrombin on (86)Rb uptake. Added alone, PD145065 did not alter (86)Rb uptake. After an incubation period of 30 minutes, thrombin increased the concentration of ET-1 in the bathing medium in a dose-dependent manner. The time course of ET-1 appearance in the bathing medium of thrombin-treated lenses showed a peak at 30 minutes followed by a gradual decline. Consistent with the idea that release of ET-1 from the lens is tightly regulated, neither the calcium ionophore A23187 (1 microM) nor depolarization by potassium-rich solution caused significant release. However, exposing the lens to insulin (150 nM) significantly increased the appearance of ET-1 in the bathing medium. In parallel studies, mRNA for prepro-ET-1 was detected in the epithelium of freshly isolated lenses. CONCLUSIONS: The results of the study suggest that ET-1 is produced in porcine lens cells and that thrombin and insulin are capable of stimulating the release of ET-1 from the lens. Thrombin-induced inhibition of Na,K-ATPase-dependent ion transport may be mediated in part through the activation of ET-1 receptors by ET-1 released from the lens. PMID- 11867601 TI - Accumulation of topically applied porcine insulin in the retina and optic nerve in normal and diabetic rats. AB - PURPOSE: To explore the pharmacokinetics of topical insulin administration in relation to retinal and optic nerve retention. METHODS: Insulin eye drops (approximately 15 microL: 0.75% porcine insulin + 0.5% permeation enhancer) were applied to the eyes of normal and diabetic rats. The rats were killed at various intervals up to 16 hours, and the retinas and optic nerves from both eyes were analyzed for the presence of insulin in an ELISA. The extent to which systemically absorbed insulin accounted for the findings of insulin in the retina was explored by examining the effects of intravenously injected insulin on retinal insulin levels and by examining the effects of eye drop administration in decapitated rats. RESULTS: Insulin levels rose significantly and peaked in the retina of normal rats 20 minutes after eye drop application (0.7 pg/microg; P < 0.00001). Levels in diabetic retinas peaked at 60 minutes (0.66 pg/microg; P < 0.004) and remained elevated for a longer period than in normal rats. The contralateral retina showed delayed accumulation of lesser amounts of insulin in both normal and diabetic rats. Significant elevations also occurred in the optic nerves in normal and diabetic rats, with concentrations reaching 13 pg/microg in normal rats at 20 minutes and 26 pg/microg in diabetic rats at 5 hours. Topical insulin application resulted in a decrease in serum glucose concomitant with an increase in serum porcine insulin. It did not appear, however, that the systemic absorption of insulin contributed to the accumulation of insulin in the ipsilateral retinas, for two reasons: The intravenous injection of a high concentration of insulin did not appreciably influence retinal insulin levels, and the application of insulin eye drops to decapitated rats still resulted in the accumulation of insulin in the retina. CONCLUSIONS: These results led to the conclusion that topically applied insulin accumulates in the retina and optic nerve in normal and diabetic rats, with levels remaining elevated longer in diabetic animals. It did not appear that systemically absorbed insulin, resulting from ocular drainage, contributed to this effect. PMID- 11867598 TI - Effect of pituitary adenylate cyclase-activating peptide on isolated rabbit iris sphincter and dilator muscles. AB - PURPOSE: Pituitary adenylate cyclase-activating peptide (PACAP) is a sensory neuropeptide in the eye that is released by noxious stimuli and considered to be a mediator of the neurogenic ocular injury response, including miosis. The purpose of this study was to clarify the functional role of PACAP in iris sphincter and dilator muscles. METHODS: Iris sphincter and dilator muscles were isolated from rabbit eyes, and the effect of PACAP on mechanical responses of these muscles using isometric tension-recording methods was investigated. RESULTS: The iris sphincter responded to electric field stimulation with contractions composed of fast twitch and subsequent slow components. Both PACAP 27 and PACAP 38 enhanced the twitch response, but neither had an effect on the slow response. The effect of both PACAPs on the twitch response was dose dependent. Neither PACAP had an effect on the amplitude of contraction evoked by exogenously applied Ach. For the iris dilator muscle, PACAP 27 inhibited the contractions induced by field stimulation or phenylephrine, whereas PACAP 38 had no effect. CONCLUSIONS: Both PACAP 27 and PACAP 38 enhance cholinergic transmission in sphincter muscle. The PACAP 27 induces relaxation of the dilator muscle by a direct effect on the muscle itself. The PACAP released during an ocular inflammatory response may induce miosis by the enhancement of cholinergic stimulation of the iris sphincter and by direct relaxation of the dilator muscles. PMID- 11867602 TI - Altered retinal function and structure after chronic placental insufficiency. AB - PURPOSE: To consider whether growth restriction secondary to chronic placental insufficiency results in postnatal deficits in retinal structure and function. METHODS: Chronic placental insufficiency was induced just before midgestation in guinea pigs through unilateral ligation of the uterine artery. Eight weeks after birth, electroretinograms were recorded from prenatally compromised (PC, n = 6) and control (n = 15) animals. Data were collected for b-wave amplitude and implicit time, also the modeled receptoral (P3) response and oscillatory potentials were extracted. After electroretinography, retinas were prepared for structural analysis (PC, n = 6; control, n = 7). A separate cohort of PC (n = 8) and control (n = 9) animals underwent tyrosine hydroxylase immunoreactivity (TH IR, dopaminergic neurons) and nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) histochemistry (neuronal nitric oxide synthase, nNOS)--these being markers of amacrine cell subpopulations. RESULTS: Electroretinography revealed two PC guinea pigs with marked changes to saturated receptoral amplitude (Rm(P3)), sensitivity (log S) and postreceptoral waveforms. Grouped PC data revealed significantly reduced Rm(P3), whereas log S was not affected. The b-wave amplitudes were normal, but b-wave implicit times were delayed (P < 0.05) in PC animals. Amplitudes and peak times of oscillatory potentials were also significantly reduced and delayed (P < 0.05). Morphologic analysis revealed significant reductions in all cellular and plexiform (synaptic) layers in both the central (P < 0.05) and peripheral (P < 0.05) retina in PC animals. The outer retina, which contains the photoreceptors and the outer plexiform layer was particularly affected. The reduced growth of plexiform layers suggests a reduction in the growth of the neuropile in PC animals compared with control animals. The total number (P < 0.03) and density (P < 0.05) of TH-IR neurons was reduced, whereas the total number and density of nNOS-positive amacrine cells was not significantly different between PC and control animals. CONCLUSIONS: Chronic placental insufficiency results in morphologic and functional alterations to the retina. Electroretinogram deficits in PC animals indicated both inner and outer retinal anomalies. Such affects could contribute to the visual impairments reported in very-low-birth-weight children, some of whom are growth restricted. PMID- 11867603 TI - Long-term effects of light damage on the retina of albino and pigmented rats. AB - PURPOSE: To observe the morphology and physiology of the retina in rats 11 weeks after a constant (24-hour) but moderate (500-lux) illumination for 1 week. METHODS: Levels of aspartate, gamma-aminobutyric acid (GABA), glutamate, glutamine, and taurine were measured by high-pressure liquid chromatography (HPLC) in the retina and vitreous humor of albino (Wistar) and pigmented (Long Evans) rats. Semithin sections were used to determine retinal morphology. The TUNEL method was used to detect cells degenerating by apoptosis. Because the GABAergic system has been shown to be particularly sensitive to the loss of photoreceptors, an additional immunohistochemical study using anti-GABA, anti glutamate decarboxylase (GAD)(67) and anti-GAD(65) antibodies was performed. RESULTS: No apparent morphologic changes were found in the retina of pigmented rats after constant illumination, whereas in albino rats disappearance of photoreceptors (except in the extreme retinal periphery) and cell bodies was observed. A significant number of TUNEL-positive nuclei also occurred in the remaining nuclear and ganglion cell layers. However, no change in the distribution of GABA, GAD(67), and GAD(65) immunoreactivities was found in either strain under constant illumination compared with control animals. Constant illumination affected the retinal levels of aspartate, glutamate, glutamine, glycine in both strains, whereas GABA contents did not change and taurine was decreased only in albino rats. A significant increase of vitreal glutamate levels was also found in both strains and of taurine levels only in albino rats. CONCLUSIONS: Phototoxicity can provoke durable retinal alterations beyond the period of lighting, suggesting progressive and probably continuous modifications of retinal physiology, even in pigmented animals in which the retina seems morphologically normal. PMID- 11867604 TI - Pigment epithelium-derived factor suppresses ischemia-induced retinal neovascularization and VEGF-induced migration and growth. AB - PURPOSE: To determine the effect of pigment epithelium-derived factor (PEDF) in a mouse model of ischemia-induced retinal neovascularization and on vascular endothelial growth factor (VEGF)--induced migration and growth of cultured microvascular endothelial cells. METHODS: Human recombinant PEDF was expressed in the human embryonic kidney 293 cell line and purified by ammonium sulfate precipitation and cation exchange chromatography. C57BL/6 mice were exposed to 75% oxygen from postnatal day (P)7 to P12 and then returned to room air. Mice received intravitreal injections of 2 microg PEDF in one eye and vehicle in the contralateral eye on P12 and P14. At P17, mice were killed and eyes enucleated for quantitation of retinal neovascularization. The mitogenic and motogeneic effects of VEGF on cultured bovine retinal and adrenal capillary endothelial cells were examined in the presence or absence of PEDF, using cell counts and migration assays. RESULTS: Two species of human recombinant PEDF, denoted A and B, were purified to apparent homogeneity. PEDF B appeared to comigrate on SDS PAGE with PEDF from human vitreous samples. Changes in electrophoretic mobility after peptide-N-glycosidase F (PNGase F) digestion suggest that both PEDF forms contain N-linked carbohydrate. Analyses of the intact proteins by liquid chromatography-electrospray mass spectrometry (LC-ESMS) revealed the major molecular weight species for PEDF A (47,705 +/- 4) and B (46,757 +/- 5). LC-ESMS analysis of tryptic peptides indicated that PEDF A and B exhibit differences in glycopeptides containing N-acetylneuraminic acid (NeuAc) and N-acetylhexosamine (HexNAc). Intravitreal administration of either species of PEDF significantly inhibited retinal neovascularization (83% for PEDF A and 55% for PEDF B; P = 0.024 and 0.0026, respectively). PEDF A and B (20 nM) suppressed VEGF-induced retinal microvascular endothelial cell proliferation by 48.8% and 41.4%, respectively, after 5 days (P < 0.001) and VEGF-induced migration by 86.5% +/- 16.7% and 78.1% +/- 22.3%, respectively, after 4 hours (P = 0.004 and P = 0.008, respectively). CONCLUSIONS: These data indicate that elevated concentrations of PEDF inhibit VEGF-induced retinal endothelial cell growth and migration and retinal neovascularization. These findings suggest that localized administration of PEDF may be an effective approach for the treatment of ischemia-induced retinal neovascular disorders. PMID- 11867605 TI - Photodynamic effects on choroidal neovascularization and physiological choroid. AB - PURPOSE: To evaluate the effect of photodynamic therapy (PDT) on perfusion and vascular integrity of choroidal neovascularization (CNV) and collateral physiological choroid. METHODS: In a prospective clinical trial, patients with subfoveal CNV were treated with PDT and verteporfin. Indocyanine green angiography (ICG-A), using a confocal laser scanning system with tomographic sections, was performed continuously 1 week before and 1, 4, and 12 weeks after and a mean long-term follow-up of 16.5 months after the final PDT. Vascular changes were localized tomographically and quantified on the level of the CNV and collateral choroid according to early lesion size, late hyperfluorescence, and persistence or recurrence. Data were analyzed separately from 38 eyes in a single and 12 eyes in a multiple-treatment regimen. RESULTS: CNV lesions were significantly reduced in size and late hyperfluorescence. However, 54% of lesions primarily demonstrated persistence, typically of the choroidal feeding complex, which was only detectable by ICG-A. Regrowth from the feeding vessel occurred regularly, but did not reach baseline dimensions. Collateral choroid exposed to photoactivation exhibited choriocapillary occlusion. Progressive recanalization was documented within 4 to 12 weeks after both single and multiple PDT. Residual changes in the choroidal filling pattern often persisted during long-term follow up. CONCLUSIONS: Tomographic ICG-A after PDT reveals persistence of CNV and/or the feeder vessel and a reduction in perfusion within the entire photosensitized area, including the surrounding choroid. Repair mechanisms occur slowly in neovascular and normal choroidal structures. PMID- 11867606 TI - Targeting of interferon to choroidal neovascularization by use of dextran and metal coordination. AB - PURPOSE: Bioactive proteins such as interferon (IFN) have been reported to be combined with water-soluble polymers, such as dextran, through metal coordination, without need for complicated procedures. In the current study, the targeting and inhibitory effects of IFN combined with dextran on experimental choroidal neovascularization (CNV) were studied in vivo. METHODS: Interferon (IFN)beta was conjugated to dextran, which has metal-chelating, diethylenetriaminepentaacetic acid (DTPA) residues. Based on metal coordination, conjugation of IFNbeta with DTPA-dextran resulted from simply mixing both substances in an aqueous solution containing Zn(2+). The effects of IFNbeta on the proliferation of human umbilical vein endothelial cells (HUVECs) and bovine retinal pigment epithelial cells (BRPECs) were evaluated. To evaluate the activity loss of IFNbeta by conjugation, the effect of the conjugate on HUVECs was compared with that of free IFNbeta. Experimental CNV was induced by subretinal injection of gelatin microspheres containing basic fibroblast growth factor in rabbits. The rabbits with CNV were intravenously treated twice weekly with 7.5 million international units (MIU)/kg per day free IFNbeta (for 4 weeks), with IFNbeta-DTPA-dextran conjugate containing 7.5 (for 2 weeks) or 0.75 (for 4 weeks) MIU/kg per day IFNbeta, or with saline. The effects of these substances were evaluated by fluorescein angiography and histology. To observe the accumulation of conjugate, the doses of IFNbeta in CNV tissues were measured by enzyme-linked immunosorbent assay. RESULTS: IFNbeta inhibited the growth of HUVECs and enhanced the proliferation of BRPECs. The conjugate seemed to preserve approximately 44% of IFNbeta activity. Although both doses of IFNbeta-DTPA dextran inhibited progression of CNV in rabbits, longer term administration of a lower dose of IFNbeta-DTPA-dextran had a sustained inhibitory effect on progression of CNV (P < 0.05). Histologic studies revealed the inhibitory effect of IFNbeta-DTPA-dextran on progression of CNV. This conjugate prolonged the plasma half-life of IFNbeta and enabled IFNbeta to accumulate in the CNV in rabbits. CONCLUSIONS: In this study, human IFNbeta was successfully used to target CNV, an enhanced antiangiogenic effect was achieved by combining it with dextran, based on metal coordination. This targeted delivery of IFNbeta may have potential as a treatment modality for CNV. PMID- 11867607 TI - Altered expression patterns of VEGF receptors in human diabetic retina and in experimental VEGF-induced retinopathy in monkey. AB - PURPOSE: The vascular endothelial growth factor (VEGF) family is involved in vascular leakage and angiogenesis in diabetic retinopathy (DR) in the eye, but may also have physiological functions. Based on the hypothesis that differential VEGF receptor (VEGFR) expression in the retina is an important determinant of effects of VEGF, this study was conducted to investigate VEGFR expression in the diabetic retina and in an experimental monkey model of VEGF-A-induced retinopathy. METHODS: In retinas of 27 eyes of diabetic donors, 18 eyes of nondiabetic control donors, and 4 monkey eyes injected with PBS or VEGF-A, expression patterns of VEGFR-1, -2, and -3 in relation to leaky microvessels, as identified by the marker pathologische anatomie Leiden-endothelium (PAL-E) were studied by immunohistochemistry. RESULTS. In control human retinas and retinas of PBS-injected monkey eyes, all three VEGFRs were expressed in nonvascular areas, but only VEGFR-1 was constitutively expressed in retinal microvessels. In diabetic eyes, increased microvascular VEGFR-2 expression was found in association with PAL-E expression, whereas microvascular VEGFR-3 was present in a subset of PAL-E-positive cases. In VEGF-A-injected monkey eyes, VEGFR-1, -2, and 3 and PAL-E were expressed in retinal microvessels. CONCLUSIONS: The VEGFR-1, -2, and -3 expression patterns in control retinas suggest physiological functions of VEGFs that do not involve the vasculature. Initial vascular VEGF signaling may act primarily through VEGFR-1. In diabetic eyes, expression of retinal VEGFR-2 and -3 is increased, mainly in leaky microvessels, and VEGF-A induces vascular expression of the VEGF-A receptor VEGFR-2 and the VEGF-C/D receptor VEGFR-3. These findings indicate a dual role of VEGFs in the physiology and pathophysiology of the retina and suggest that microvascular VEGFR-2 and -3 signaling by VEGFs occurs late in the pathogenesis of DR, possibly initiated by high levels of VEGF-A in established nonproliferative DR. PMID- 11867608 TI - Distribution of organic anion-transporting polypeptide 2 (oatp2) and oatp3 in the rat retina. AB - PURPOSE: To examine the expression of multifunctional Na(+)-independent organic anion-transporting polypeptides, termed oatp1, oatp2, and oatp3, involving the transport of thyroid hormone in the rat retina at the protein and mRNA levels. METHODS: Northern blot analysis was performed using oatp1, -2, and -3 cDNAs. Reverse transcription-polymerase chain reaction (RT-PCR) was also performed using gene-specific primers for oatp1, -2, and -3. mRNA distribution of these oatps in the rat retina was examined by in situ hybridization. Western blot analysis and immunohistochemistry were also performed by raising specific antibodies against oatp2 and -3. RESULTS: Northern blot analysis showed that the mRNAs for oatp2 and -3 were expressed in the rat retina and retinal pigment epithelium (RPE). Amplified cDNA products by RT-PCR for oatp2 and -3 were also detected in the rat retina-RPE. In contrast, no specific band for oatp1 was detected by Northern blot analysis or RT-PCR. By in situ hybridization, oatp2-specific mRNA signals were seen in the RPE and inner nuclear layer, whereas the oatp3 mRNA signal was localized to the ganglion cell. At the protein level, a single band for oatp2 and -3 proteins was detected in the rat retina-RPE by Western blot analysis. Immunohistochemistry revealed that oatp2 immunostaining was predominantly expressed at the apical surface of the RPE. Weak immunostaining for oatp2 was also seen in the inner nuclear layer and the ganglion cell layer. In contrast, apparent immunostaining for oatp3 was seen in the nerve fiber layer, ganglion cell layer, inner plexiform layer, and outer aspect of the inner nuclear layer. In addition, oatp3 immunostaining was detected predominantly in the optic nerve fiber. CONCLUSIONS: These results reveal that oatp2 is localized mainly in the RPE, suggesting a role for organic anion transport in this specialized ocular tissue. In contrast, oatp3 is localized mainly in optic nerve fibers, suggesting that oatp3 is a specific transporter in the visual nervous system. In conclusion, these data suggest that oatp2 and -3 may be involved in the transport of thyroid hormone in the rat retina. PMID- 11867610 TI - Upregulation of extracellular ATP-induced Muller cell responses in a dispase model of proliferative vitreoretinopathy. AB - PURPOSE: To test whether in an animal model of proliferative vitreoretinopathy (PVR) the Muller glial cells displayed an upregulation of purinergic P2 receptor mediated responses. METHODS: PVR was induced by intravitreal injection of the proteolytic enzyme, dispase, in the eyes of adult rabbits. The developing PVR was examined ophthalmoscopically. After 3 weeks, small retinal pieces were wholemounted and used for calcium imaging, freshly dissociated Muller cells were subjected to calcium imaging, and patch-clamp recordings were made. The presence of P2 receptor-mediated Ca(2+) responses was determined both directly--that is, fluorometrically--and indirectly, by electrophysiological recording of Ca(2+) activated K(+) currents. RESULTS: According to earlier observations in another model of retinal detachment and PVR, the reactive Muller cells displayed hypertrophy, downregulation of inwardly rectifying K(+) currents, and depolarization of the resting membrane potential, all dependent on the severity of the PVR. Further, significant PVR-induced increase was observed in the number of Muller cells responding to adenosine 5'-triphosphate (ATP), with a transient elevation of their [Ca(2+)](i). If isolated Muller cells were exposed to ATP, 13% of the control cells, but 29% (moderate PVR) or 53% (massive PVR) of the reactive cells, showed fluorometric Ca(2+) increases. An increase of Ca(2+)-activated K(+) currents was measured in 11% of the control cells, but in 83% (moderate PVR) and 90% (massive PVR) of the reactive cells. Confocal images of retinal wholemounts revealed similar results. Because similar responses were elicited by uridine triphosphate (UTP), the dominant involvement of metabotropic (P2Y type) purinergic receptors is suggested. CONCLUSIONS: An upregulation of purinergic receptors is part of the reactive changes of Muller cells during PVR. It is suggested that ATP-evoked Ca(2+) responses may support the proliferation of Muller cells during PVR. PMID- 11867609 TI - In vitro characterization of a spontaneously immortalized human Muller cell line (MIO-M1). AB - PURPOSE: To characterize a spontaneously immortalized human Muller cell line and to determine whether it retains the characteristics of primary isolated cells without undergoing differentiation in vitro. METHODS: An immortalized cell line obtained from human retina was investigated for the expression of known markers of Muller cells, including cellular retinaldehyde binding protein (CRALBP), glutamine synthetase, epidermal growth factor receptor (EGF-R), alpha-smooth muscle actin (alpha-SMA), and glial fibrillary acidic protein (GFAP). Also examined were the morphologic features of these cells, by scanning and transmission electron microscopy, and their functional characteristics, by electrogenic responses to glutamate. In addition, comparative studies were made of these cells with primary cultures of freshly isolated human Muller cells. RESULTS: The cells expressed CRALBP, EGF-R, glutamine synthetase, and alpha-SMA, as judged by confocal microscopy and Western blot analysis of cell lysates. Western blot analysis did not detect GFAP in cell lysates, but confocal microscopy showed that occasional cells expressed GFAP after detachment from the monolayer. The morphologic features of the cells examined, as judged by scanning and transmission electron microscopy, resemble those of cells derived from primary cell cultures. They possess villous projections on their apical surfaces and contain loose bundles of microtubules aligned parallel to one another and the long axis of the cell process. Characteristically, they contain abundant deposits of glycogen particles that do not differ from those seen in primary isolated cells. Preliminary recordings with intracellular electrodes revealed that these cells have properties similar to those described for mammalian Muller cells and depolarize in response to L-glutamate without significant change in membrane resistance, consistent with the well-established electrogenic uptake of this amino acid. CONCLUSIONS: A spontaneously immortalized Muller cell line was characterized that retains the characteristics of primary isolated cells in culture. To the authors' knowledge, it constitutes the first human Muller cell line reported in the literature. It has been named MIO-M1 (Moorfields/Institute of Ophthalmology-Muller 1) after the authors' institution. Availability of this human cell line will facilitate studies designed to obtain a better understanding of the role of Muller cells in normal and pathologic conditions. PMID- 11867611 TI - Endothelin-induced changes in the physiology of retinal pericytes. AB - PURPOSE: Pericytes are positioned on the abluminal wall of capillaries and are thought to play a role in regulating retinal blood flow. Although endothelin (ET) 1 is a putative endothelium-pericyte signal, the mechanisms by which this molecule regulates pericyte function remain unclear. Because ion channels play a vital role in the response of pericytes to extracellular signals, this study was undertaken to assess the effects of ET-1 on ionic currents. METHODS: The perforated-patch configuration of the patch-clamp technique was used to monitor whole-cell currents of pericytes located on microvessels freshly isolated from the rat retina. To assay cell-to-cell coupling within retinal microvessels, a gap junction--permeant tracer was loaded through patch pipettes into pericytes and the spreading of the tracer detected by immunohistochemistry. RESULTS: ET-1 acting through ET(A) receptors altered pericyte currents and caused depolarization of the membrane potential. The effects on pericyte currents were dynamic over time. Initially, the nonspecific cation (NSC) and calcium-activated chloride (Cl(Ca)) currents were activated and the adenosine triphosphate (ATP) sensitive potassium (K(ATP)) current inhibited. Subsequently, by a mechanism sensitive to a protein kinase C (PKC) inhibitor, the NSC, Cl(Ca), and voltage dependent potassium currents diminished as gap junction pathways closed within the microvessels. CONCLUSIONS: ET-1 regulates pericyte conductances by multiple mechanisms. One process involves a PKC-dependent closure of gap junction pathways resulting in loss of electrotonic input from neighboring cells. Thus, ET-1 not only affects individual microvascular cells, but also regulates the effective size of the multicellular functional units that may serve to control capillary blood flow. This regulation of intercellular communication within pericyte containing microvessels may be an important, previously unrecognized, action of ET-1. PMID- 11867612 TI - MT(1) melatonin receptor in the human retina: expression and localization. AB - PURPOSE: Melatonin's function in human vision is far from understood, in part because of the lack of information on its cellular targets. Therefore, expression and localization of the MT(1) melatonin receptor in human retina was examined. METHODS: Postmortem nonpathologic human eyes from nine donors were investigated, three by reverse transcription-polymerase chain reaction (RT-PCR) for MT(1) and MT(2) transcripts and six by immunocytochemistry, using a peptide-specific anti MT(1) receptor antibody. RESULTS: RT-PCR suggested that both MT(1) and MT(2) transcripts had similar levels of expression. Vertical slices of human retina demonstrated MT(1) immunoreactivity in cell bodies along the outer border of the inner nuclear layer (INL), along the inner border of the INL, in cell bodies within the ganglion cell layer (GCL), and in the inner segments (IS) of photoreceptors. Double immunolabeling using anti-MT(1) and tyrosine hydroxylase revealed that 69% of CA1 and 63% of CA2 dopaminergic neurons exhibited MT(1) immunoreaction. Double immunolabeling with anti-parvalbumin, a horizontal cell marker, showed that MT(1)-positive cells along the outer INL border were exclusively horizontal cells, and that 18% of horizontal cells in central retina expressed MT(1). Double staining with MT(1) and markers for both rod and cone photoreceptors suggest that IS staining is present on rod cells. CONCLUSIONS: The MT(1) receptor is expressed in diverse neuronal cell types in the human retina, providing evidence of a significant role for melatonin and this receptor subtype in human vision. PMID- 11867613 TI - Cone-specific mediation of rod sensitivity in trichromatic observers. AB - PURPOSE: The slope of the rod threshold versus the illuminance (TVI) function changes with the wavelength of the background light. This study was conducted to determine whether the changes in slope are due to the stimulation of specific cone classes. METHODS: An eight-channel optical system was used to generate lights that differed in cone and rod photoreceptor illuminance. Rod flicker TVI functions were measured in normal trichromatic observers at mesopic light levels. The independent variables were (1) the relative contribution of the short (S)- and long (L)- wavelength cones to the background light (i.e., the background lights varied along S-only and L-only lines), and (2) the temporal frequency of the flickering lights (4, 7.5, and 15 Hz). RESULTS: The 4-Hz rod flicker TVI function had a slope of 0.87 when measured near W (MacLeod-Boynton chromaticity of 0.66, 1.0). At 4 and 7.5 Hz, an increase in the relative L-cone illuminance steepened the slope of the rod-only TVI curve, but an increase in the relative S cone illuminance had no effect. The slope of the 7.5-Hz TVI function decreased at higher illuminance levels. At 15 Hz, the thresholds could be measured over only a limited range. CONCLUSIONS: The L-cone system contributes to the desensitization of the rod system at mesopic light levels, whereas, in the range of lights used in these experiments, the S-cone system apparently does not. The possibility that S-cone stimulation desensitizes the response to rod signals at higher levels of S cone illumination cannot be eliminated. PMID- 11867614 TI - DNA binding and gene activation properties of the Nmp4 nuclear matrix transcription factors. AB - Splice variants of the Nmp4 gene include nuclear matrix transcription factors that regulate the type I collagen alpha1(I) polypeptide chain (COL1A1) promoter and several matrix metalloproteinase (MMP) genes. To date, these are the only Cys(2)His(2) zinc finger proteins known to bind within the minor groove of homopolymeric (dA.dT) DNA. Nmp4 isoforms contain from 5 to 8 Cys(2)His(2) zinc fingers, an SH3-binding domain that overlaps with a putative AT-hook and a polyglutamine-alanine repeat (poly(QA)). To determine the mechanistic significance of Cys(2)His(2) zinc finger association with this unusual consensus DNA binding element, we identified the Nmp4 DNA-binding and transcriptional activation domains. Zinc fingers 2, 3, and 6 mediated association with the homopolymeric (dA.dT) COL1A1/MMP DNA consensus element. The N terminus of the Nmp4 protein exhibited a strong trans-activation capacity when fused to the GAL4 DNA-binding domain, but this activity was masked within the context of the full length Nmp4-GAL4 DNA-binding domain chimera. However, upon binding to the COL1A1/MMP homopolymeric (dA.dT) element, the native Nmp4 protein up-regulated transcription, and the poly(QA) domain acquired a significant role in trans activation. We propose that allosteric effects induced upon zinc finger association with the homopolymeric (dA.dT) minor groove confer context-specific functionality to this unusual family of Cys(2)His(2) transcription factors. PMID- 11867615 TI - Codon-anticodon interaction at the P site is a prerequisite for tRNA interaction with the small ribosomal subunit. AB - The arrival of high resolution crystal structures for the ribosomal subunits opens a new phase of molecular analysis and asks for corresponding analyses of ribosomal function. Here we apply the phosphorothioate technique to dissect tRNA interactions with the ribosome. We demonstrate that a tRNA bound to the P site of non-programmed 70 S ribosomes contacts predominantly the 50 S, as opposed to the 30 S subunit, indicating that codon-anticodon interaction at the P site is a prerequisite for 30 S binding. Protection patterns of tRNAs bound to isolated subunits and programmed 70 S ribosomes were compared. The results suggest the presence of a movable domain in the large ribosomal subunit that carries tRNA and reveal that only approximately 15% of a tRNA, namely residues 30 +/- 1 to 43 +/- 1, contact the 30 S subunit of programmed 70 S ribosomes, whereas the remaining 85% make contact with the 50 S subunit. Identical protection patterns of two distinct elongator tRNAs at the P site were identified as tRNA species independent phosphate backbone contacts. The sites of protection correlate nicely with the predicted ribosomal-tRNA contacts deduced from a 5.5-A crystal structure of a programmed 70 S ribosome, thus refining which ribosomal components are critical for tRNA fixation at the P site. PMID- 11867616 TI - Calcium influx factor from cytochrome P-450 metabolism and secretion-like coupling mechanisms for capacitative calcium entry in corneal endothelial cells. AB - Notwithstanding extensive efforts, the mechanism of capacitative calcium entry (CCE) remains unclear. Two seemingly opposed theories have been proposed: secretion-like coupling (Patterson, R. L., van Rossum, D. B., and Gill, D. L. (1999) Cell 98, 487-499) and the calcium influx factor (CIF) (Randriamampita, C., and Tsien, R. Y. (1993) Nature 364, 809-814). In the current study, a combinatorial approach was taken to investigate the mechanism of CCE in corneal endothelial cells. Induction of cytochrome P-450s by beta-naphthoflavone (BN) enhanced CCE measured by Sr(2+) entry after store depletion. 5,6 Epoxyeicosatrienoic acid (5,6-EET), a proposed CIF generated by cytochrome P-450s (Rzigalinski, B. A., Willoughby, K. A., Hoffman, S. W., Falck, J. R., and Ellis, E. F. (1999) J. Biol. Chem. 274, 175-182), induced Ca(2+) entry. Both BN-enhanced CCE and the 5,6-EET-induced Ca(2+) entry were inhibited by the CCE blocker 2 aminoethoxydiphenyl borate, indicating a role for cytochrome P-450s in CCE. Treatment with calyculin A (CalyA), which causes condensation of cortical cytoskeleton, inhibited CCE. The actin polymerization inhibitor cytochalasin D partially reversed the inhibition of CCE by CalyA, suggesting a secretion-like coupling mechanism for CCE. However, CalyA could not inhibit CCE in BN-treated cells, and 5,6-EET caused a partial activation of CCE in CalyA-treated cells. These results further support the notion that cytochrome P-450 metabolites may be CIFs. The vesicular transport inhibitor brefeldin A inhibited CCE in both vehicle and BN-treated cells. Surprisingly, Sr(2+) entry in the absence of store depletion was enhanced in BN-treated cells, which was also inhibited by 2 aminoethoxydiphenyl borate. An integrative model suggests that both CIF from cytochrome P-450 metabolism and secretion-like coupling mechanisms play roles in CCE in corneal endothelial cells. PMID- 11867618 TI - Transcriptional activation of cytochrome P450 CYP2C45 by drugs is mediated by the chicken xenobiotic receptor (CXR) interacting with a phenobarbital response enhancer unit. AB - Cytochromes P450 (CYP)-2C enzymes fulfill an important role in xenobiotic metabolism and therefore have extensively been studied in rodents and humans. However, no CYP2C genes have been described in avian species to date. In this paper, we report the cloning, functional analysis, and regulation of chicken CYP2C45. The sequence shares up to 58% amino acid identity with CYP2Cs in other species. The overexpression of CYP2C45 in chicken hepatoma cells leghorn male hepatoma (LMH) led to increased scoparone metabolism. CYP2C45 regulation was studied in LMH cells at the mRNA level and in reporter gene assays using a construct containing 2.6 kb of its 5'-flanking region. Exposure of LMH cells to phenobarbital or metyrapone led to a 95- or 210-fold increase in CYP2C45 mRNA and a 140- or 290-fold increase in reporter gene expression, respectively. A phenobarbital response enhancer unit (PBRU) of 239 bp containing a DR-4 nuclear receptor binding site was identified within the 2.6-kb fragment. Site-specific mutation of the DR-4 revealed the requirement of this motif for CYP2C45 induction by drugs. The chicken xenobiotic receptor CXR interacted with the PBRU in electromobility shift and transactivation assays. Furthermore, the related nuclear receptors, mouse PXR and mouse CAR, transactivated this enhancer element, suggesting evolutionary conservation of nuclear receptor-DNA interactions in CYP2C induction. PMID- 11867617 TI - Analysis of tissue transglutaminase function in the migration of Swiss 3T3 fibroblasts: the active-state conformation of the enzyme does not affect cell motility but is important for its secretion. AB - Increasing evidence suggests that tissue transglutaminase (tTGase; type II) is externalized from cells, where it may play a key role in cell attachment and spreading and in the stabilization of the extracellular matrix (ECM) through protein cross-linking. However, the relationship between these different functions and the enzyme's mechanism of secretion is not fully understood. We have investigated the role of tTGase in cell migration using two stably transfected fibroblast cell lines in which expression of tTGase in its active and inactive (C277S mutant) states is inducible through the tetracycline-regulated system. Cells overexpressing both forms of tTGase showed increased cell attachment and decreased cell migration on fibronectin. Both forms of the enzyme could be detected on the cell surface, but only the clone overexpressing catalytically active tTGase deposited the enzyme into the ECM and cell growth medium. Cells overexpressing the inactive form of tTGase did not deposit the enzyme into the ECM or secrete it into the cell culture medium. Similar results were obtained when cells were transfected with tTGase mutated at Tyr(274) (Y274A), the proposed site for the cis,trans peptide bond, suggesting that tTGase activity and/or its tertiary conformation dependent on this bond may be essential for its externalization mechanism. These results indicate that tTGase regulates cell motility as a novel cell-surface adhesion protein rather than as a matrix cross-linking enzyme. They also provide further important insights into the mechanism of externalization of the enzyme into the extracellular matrix. PMID- 11867619 TI - Dual promoter structure of mouse and human fatty acid translocase/CD36 genes and unique transcriptional activation by peroxisome proliferator-activated receptor alpha and gamma ligands. AB - Fatty acid translocase (FAT)/CD36 is a glycoprotein involved in multiple membrane functions including uptake of long-chain fatty acids and oxidized low density lipoprotein. In mice, expression of the gene is regulated by peroxisome proliferator-activated receptor (PPAR) alpha in the liver and by PPAR gamma in the adipose tissues (Motojima, K., Passilly, P. P., Peters, J. M., Gonzalez, F. J., and Latruffe, N. (1998) J. Biol. Chem. 273, 16710-16714). However, the time course of PPAR alpha ligand-induced expression of FAT/CD36 in the liver, and also in the cultured hepatoma cells, is significantly slower than those of other PPAR alpha target genes. To study the molecular mechanism of the slow transcriptional activation of the gene by a PPAR ligand, we first cloned the 5' ends of the mRNA and then the mouse gene promoter region from a genomic bacterial artificial chromosome library. Sequencing analyses showed that transcription of the gene starts at two initiation sites 16 kb apart and splicing occurs alternatively, producing at least three mRNA species with different 5'-noncoding regions. The PPAR alpha ligand-responsive promoter in the liver was identified as the new upstream promoter where we found several possible binding sites for lipid metabolism-related transcriptional factors but not for PPAR. Neither promoter responded to a PPAR alpha ligand in the in vitro or in vivo reporter assays using cultured hepatoma cells and the liver of living mice. We also have cloned the human FAT/CD36 gene from a bacterial artificial chromosome library and identified a new independent promoter that is located 13 kb upstream of the previously reported promoter. Only the upstream promoter responded to PPAR alpha and PPAR gamma ligands in a cell type-specific manner. The absence of PPRE in the responding upstream promoter region, the delayed activation by the ligand, and the results of the reporter assays all suggested that transcriptional activation of the FAT/CD36 gene by PPAR ligands is indirectly dependent on PPAR. PMID- 11867621 TI - Aspartic acid 564 in the third cytoplasmic loop of the luteinizing hormone/choriogonadotropin receptor is crucial for phosphorylation-independent interaction with arrestin2. AB - Arrestin2 binding to the active but unphosphorylated luteinizing hormone/choriogonadotropin receptor (LH/CG R) in ovarian follicles is triggered by activation of ADP-ribosylation factor 6 (ARF6) and leads to uncoupling of this receptor from cAMP signaling. We sought to determine how arrestin2 binds to LH/CG R, if binding is of high affinity, and if the receptor also binds arrestin3. Desensitization of intact LH/CG R was equally sensitive to ectopic constructs of arrestin2 that bind other G protein-coupled receptors (GPCRs) either in a phosphorylation-independent or -dependent manner. Intact LH/CG R was not desensitized by ectopic arrestin3 constructs. Surface plasmon resonance studies showed that arrestin2 bound a synthetic third intracellular (3i) LH/CG R loop peptide with picomolar affinity; arrestin3 bound with millimolar affinity. To determine whether Asp-564 in the 3i loop mimicked the phosphorylated residue of other GPCRs, human embryonic kidney (HEK) cells were transfected with wild-type (WT) and D564G LH/CG R. An agonist-stimulated ARF6-dependent arrestin2 undocking pathway to drive desensitization of WT receptor was recapitulated in HEK cell membranes, and ectopic arrestin2 promoted desensitization of WT LH/CG R. However, D564G LH/CG R in HEK cells was not desensitized, and synthetic 3i D564G peptide did not bind arrestin2. Synthetic 3i loop peptides containing D564E, D564V, or D564N also did not bind arrestin2. We conclude that the ARF6-mediated mechanism to release a pool of membrane-delimited arrestin to bind GPCRs may be a widespread mechanism to deliver arrestin to GPCRs for receptor desensitization. Unlike other GPCRs that additionally require receptor phosphorylation, LH/CG R activation is sufficient to expose a conformation in which Asp-564 in the 3i loop confers high affinity binding selectively to arrestin2. PMID- 11867620 TI - RhoA and Rho kinase-dependent phosphorylation of moesin at Thr-558 in hippocampal neuronal cells by glutamate. AB - When we were studying phosphorylated proteins in the rat brain after electroconvulsive shock (ECS), we observed the rapid phosphorylation of a 75-kDa protein, which cross-reacted with the anti-phospho-p70 S6 kinase antibody. The phosphorylated protein was purified and identified as moesin, a member of the ezrin/radixin/moesin (ERM) family and a general cross-linker between cortical actin filaments and plasma membranes. The purified moesin from rat brain was phosphorylated at serine and threonine residues. Moesin was rapidly phosphorylated at the threonine 558 residue after ECS in the rat hippocampus, peaked at 1 min, and returned to the basal level by 2 min after ECS. To investigate the mechanism of moesin phosphorylation in neuronal cells, we stimulated a rat hippocampal progenitor cell, H19-7/IGF-IR, with glutamate, and observed the increased phosphorylation of moesin at Thr-558. Glutamate transiently activated RhoA, and constitutively active RhoA increased the basal level phosphorylation of moesin. The inhibition of RhoA and its effector, Rho kinase, abolished increased Thr-558 phosphorylation by glutamate in H19-7/IGF-IR cells, suggesting that the phosphorylation of moesin at Thr-558 in H19-7/IGF-IR cells by glutamate is mediated by RhoA and Rho kinase activation. PMID- 11867622 TI - Cytochrome P450 2C9-induced endothelial cell proliferation involves induction of mitogen-activated protein (MAP) kinase phosphatase-1, inhibition of the c-Jun N terminal kinase, and up-regulation of cyclin D1. AB - Cytochrome P450 (CYP)-derived epoxyeicosatrienoic acids (EETs) are important modulators of endothelial cell homeostasis. We investigated the signaling pathway linking the activation of CYP 2C9 to enhanced endothelial cell proliferation. Overexpression of CYP 2C9 in cultured human endothelial cells markedly increased proliferation. This effect was paralleled by an up-regulation of the G(1) phase regulatory protein, cyclin D1. The specific CYP 2C9 inhibitor, sulfaphenazole, prevented both the enhanced cell proliferation and up-regulation of cyclin D1. CYP 2C9 overexpression also decreased the activity of the c-Jun N-terminal kinase (JNK). Coexpression of wild type JNK with CYP 2C9 attenuated the CYP 2C9-induced increase in cyclin D1 expression and abolished the CYP 2C9-induced proliferation response. In contrast, cotransfecting dominant negative JNK with CYP 2C9 restored the CYP 2C9-mediated up-regulation of cyclin D1 and proliferation. The inactivation of JNK is linked to its dephosphorylation by dual specificity mitogen-activated protein (MAP) kinase phosphatases (MKPs). Overexpression of CYP 2C9 significantly increased the expression of MKP-1, as did incubation with 11,12 EET. These data demonstrate that the mitogenic effect of CYP 2C9 is due to the generation of EETs, which promote the MKP-1-mediated dephosphorylation and inactivation of JNK, effects ultimately culminating in the expression of cyclin D1 and endothelial cell proliferation. PMID- 11867623 TI - Role of Sp1 and Sp3 in the nutrient-regulated expression of the human asparagine synthetase gene. AB - The human asparagine synthetase (AS) gene responds to depletion of mammalian cells for either amino acids or carbohydrates. Five specific cis-elements have been implicated: three GC boxes (GC-I, GC-II and GC-III) and two nutrient-sensing response elements (NSRE-1, -2). This study shows that all three GC boxes are required to maintain basal transcription and to obtain maximal induction of the AS gene by amino acid limitation. However, there is not complete redundancy among the three GC boxes, and there is a hierarchy of importance with regard to transcription (GC-III > GC-II > GC-I). In vitro, two GC boxes formed protein-DNA complexes (GC-II and GC-III) with Sp1 and Sp3. Although transcription of the AS gene is elevated by nutrient limitation, the absolute amount of these protein-DNA complexes and the total pools of Sp1 and Sp3 did not increase. A small, but detectable portion of Sp1 was modified by phosphorylation following amino acid deprivation. In vivo, expression of Sp1 and Sp3 in Drosophila SL2 cells increased AS promoter activity. Sp1 expression increased basal transcription but did not cause a further increase when SL2 cells were amino acid-deprived. Sp3 expression enhanced both the basal and the starvation-induced transcription. PMID- 11867624 TI - Leukocyte elastase negatively regulates Stromal cell-derived factor-1 (SDF 1)/CXCR4 binding and functions by amino-terminal processing of SDF-1 and CXCR4. AB - Activation of CXCR4 by the CXC chemokine stromal cell-derived factor-1 (SDF-1) requires interaction of the amino-terminal domains of both molecules. We report that proteinases released from either mononucleated blood cells or polymorphonuclear neutrophils degranulated by inflammatory stimuli generate an SDF-1 fragment that is deleted from amino-terminal residues Lys(1)-Pro(2)-Val(3), as characterized by mass spectrometry analysis. The proteolyzed chemokine fails to induce agonistic functions and is unable to prevent the fusogenic capacity of CXCR4-tropic human immunodeficiency viruses. Furthermore, we observed that exposure of CXCR4-expressing cells to leukocyte proteinases results in the proteolysis of the extracellular amino-terminal domain of the receptor, as assessed by flow cytometry analysis and electrophoretic separation of immunoprecipitated CXCR4. Blockade of SDF-1 and CXCR4 proteolysis by the specific leukocyte elastase inhibitor, N-methoxysuccinyl-alanine-alanine-proline-valine chloromethyl ketone, identified elastase as the major enzyme among leukocyte secreted proteinases that accounts for inactivation of both SDF-1 and CXCR4. Indeed, purified leukocyte elastase generated in either SDF-1 or CXCR4 a pattern of cleavage indistinguishable from that observed with leukocyte-secreted proteinases. Our findings suggest that elastase-mediated proteolysis of SDF 1/CXCR4 is part of a mechanism regulating their biological functions in both homeostatic and pathologic processes. PMID- 11867625 TI - FOR, a novel orphan nuclear receptor related to farnesoid X receptor. AB - We have identified and characterized a new amphibian orphan member of the nuclear receptor superfamily and termed it FOR1 (farnesoid X receptor (FXR)-like Orphan Receptor) because it shares the highest amino acid identity with the mammalian FXR. We also identified a variant of FOR1, called FOR2, which has 15 additional C terminal amino acids. Both variants include an unusual insertion of 33 amino acids in the helix 7 region of the canonical ligand binding domain sequence, suggesting a unique structure for FOR. Northern blot analysis demonstrates that the FOR gene is highly expressed in adult and tadpole liver, kidney, and tail bud stage of the embryo. Detailed expression analysis using in situ hybridization indicates that FOR expression is first detectable at stage 30/31 in the presumptive liver region lasting until stage 41 with a peak level evident at stage 35/36. FOR forms heterodimeric complexes with retinoid X receptor (RXR) as demonstrated by biochemical and mammalian two-hybrid approaches. Gel mobility shift assays demonstrate that FORs form specific DNA-protein complexes on an FXR binding element consisting of an inverted repeat DNA element with 1 nucleotide spacing (IR1) from the phospholipid transfer protein gene promoter. Finally, although FORs do not exhibit constitutive transcriptional activity, frog gallbladder extract significantly augments the transcriptional activities of FORs. PMID- 11867626 TI - Folding problems of the 5' splice site containing the P1 stem of the group I thymidylate synthase intron: substrate binding inhibition in vitro and mis splicing in vivo. AB - We developed an in vitro cleaving assay for the thymidylate synthase (td) group I intron and observed that the off-rate of the substrate is faster than cleavage. From the sequence stems P1 and P2 can vary from 4 to 8 and from 6 to 10 base pairs, respectively, with folding of a long P1 stem being in competition with folding of a long P2 stem. Shorter substrates, which cannot compete with the formation of an extended P2, result in faster cleavage, suggesting that binding of the substrate indeed interferes with folding of stem P2. In vivo splicing analyses of mutants containing alterations in stems P1 and P2 indicate that the wild-type exon sequence of P1 is suboptimal for splicing. Furthermore, folding of P1 in vivo is in competition with an alternative cryptic P1 stem resulting in mis splicing. Translation promotes splicing at the correct 5' splice site, whereas in the absence of translation, mis-splicing is favored. The combination of the in vitro and in vivo assays clearly displays the folding problems for correct splice site selection in this group I intron. PMID- 11867627 TI - c-SRC mediates neurite outgrowth through recruitment of Crk to the scaffolding protein Sin/Efs without altering the kinetics of ERK activation. AB - SRC family kinases have been consistently and recurrently implicated in neurite extension events, yet the mechanism underlying their neuritogenic role has remained elusive. We report that epidermal growth factor (EGF) can be converted from a non-neuritogenic into a neuritogenic factor through moderate activation of endogenous SRC by receptor-protein-tyrosine phosphatase alpha (a physiological SRC activator). We show that such a qualitative change in the response to EGF is not accompanied by changes in the extent or kinetics of ERK induction in response to this factor. Instead, the pathway involved relies on increased tyrosine phosphorylation of, and recruitment of Crk to, the SRC substrate Sin/Efs. The latter is a scaffolding protein structurally similar to the SRC substrate Cas, tyrosine phosphorylation of which is critical for migration in fibroblasts and epithelial cells. Expression of a dominant negative version of Sin interfered with receptor-protein-tyrosine phosphatase alpha/EGF- as well as fibroblast growth factor-induced neurite outgrowth. These observations uncouple neuritogenic signaling in PC12 cells from sustained activation of ERK kinases and for the first time identify an effector of SRC function in neurite extension. PMID- 11867628 TI - p53 Activation by nitric oxide involves down-regulation of Mdm2. AB - Nitric oxide (NO) is an important bioactive molecule involved in a variety of physiological and pathological processes. At the same time, NO is also an inducer of stress signaling, owing to its ability to damage proteins and DNA. NO was reported to be a potent activator of the p53 tumor suppressor protein. However, the mechanisms underlying p53 activation by NO remain to be elucidated. We report here that NO induces the accumulation of transcriptionally active p53 in a variety of cell types and that NO signaling to p53 does not require ataxia telangiectasia-mutated (ATM), poly(ADP-ribose) polymerase 1, or the ARF tumor suppressor protein. In mouse embryonic fibroblasts, NO elicits a down-regulation of Mdm2 protein levels that precedes the rise in p53. NO-induced down-regulation of Mdm2 protein but not its mRNA also occurs in several p53-deficient cell types and is thus p53-independent. The drop in endogenous Mdm2 levels following NO treatment is accompanied by a corresponding reduction in the rate of p53 ubiquitination. Thus, the down-regulation of Mdm2 by NO is likely to contribute to the activation of p53. PMID- 11867629 TI - Catalytic properties, thiol pK value, and redox potential of Trypanosoma brucei tryparedoxin. AB - The dithiol protein tryparedoxin is a component of the unique trypanothione/trypanothione reductase metabolism of trypanosomatids and is involved in the parasite synthesis of deoxyribonucleotides and the detoxication of hydroperoxides. Tryparedoxin is a highly abundant protein in all life stages of Trypanosoma brucei, the causative agent of African sleeping sickness. As shown here, its functional properties are intermediate between those of classical thioredoxins and glutaredoxins. The redox potential of T. brucei tryparedoxin of 249 mV was determined by protein-protein redox equilibration with Escherichia coli thioredoxin. The trypanothione/tryparedoxin couple is probably the most significant factor determining the cytosolic redox potential of the parasites. The pK value of Cys(40), the first thiol in the WCPPC motif, is 7.2 as derived from the thiolate absorption at 240 nm and the rate of carboxymethylation. Alteration of the active site into that of thioredoxin (CGPC) did not affect the pK value. In contrast, in the mutant with the glutaredoxin motif (CPYC) the pK dropped to < or =4.0. The fact that the pK value of tryparedoxin coincides with the intracellular pH of the parasite may contribute to the reactivity of tryparedoxin in thiol disulfide exchange reactions. PMID- 11867630 TI - Glucocorticoids synergistically enhance nontypeable Haemophilus influenzae induced Toll-like receptor 2 expression via a negative cross-talk with p38 MAP kinase. AB - The recognition of invading microbes followed by the induction of effective innate immune response is crucial for host survival. Human surface epithelial cells are situated at host-environment boundaries and thus act as the first line of host defense against invading microbes. They recognize the microbial ligands via Toll-like receptors (TLRs) expressed on the surface of epithelial cells. TLR2 has gained importance as a major receptor for a variety of microbial ligands. In contrast to its high expression in lymphoid tissues, TLR2 is expressed at low level in epithelial cells. Thus, it remains unclear whether the low amount of TLR2 expressed in epithelial cells is sufficient for mediating bacteria-induced host defense and immune response and whether TLR2 expression can be up-regulated by bacteria during infection. Here, we show that TLR2, although expressed at very low level in unstimulated human epithelial cells, is greatly up-regulated by nontypeable Hemophilus influenzae (NTHi), an important human bacterial pathogen causing otitis media and chronic obstructive pulmonary diseases. Activation of an IKKbeta-IkappaBalpha-dependent NF-kappaB pathway is required for TLR2 induction, whereas inhibition of the MKK3/6-p38alpha/beta pathway leads to enhancement of NTHi-induced TLR2 up-regulation. Surprisingly, glucocorticoids, well known potent anti-inflammatory agents, synergistically enhance NTHi-induced TLR2 up-regulation likely via a negative cross-talk with the p38 MAP kinase pathway. These studies may bring new insights into the role of bacteria and glucocorticoids in regulating host defense and immune response and lead to novel therapeutic strategies for modulating innate immune and inflammatory responses for otitis media and chronic obstructive pulmonary diseases. PMID- 11867631 TI - Accelerating the rate of disassembly of karyopherin.cargo complexes. AB - Transport of macromolecules across the nuclear pore complex (NPC) occurs in seconds and involves assembly of a karyopherin.cargo complex and docking to the NPC, translocation of the complex across the NPC via interaction with nucleoporins (Nups), and dissociation of the complex in the nucleoplasm. To identify rate-limiting steps in the Kap95p.Kap60p-mediated nuclear import pathway of Saccharomyces cerevisiae, we reconstituted key intermediate complexes and measured their rates of dissociation and affinities of interaction. We found that a nuclear localization signal-containing protein (NLS-cargo) dissociates slowly from Kap60p monomers and Kap60p.Kap95p heterodimers with half-lives (t(12)) of 7 and 73 min, respectively; that Kap60p and Kap60p.NLS-cargo complexes dissociate slowly from Kap95p (t(12) = 36 and 73 min, respectively); and that Kap95p.Kap60p.NLS-cargo complexes and Kap95p.Kap60p heterodimers dissociate rapidly from the nucleoporin Nup1p (t(12) < or = 21 s) and other Nups. A search for factors that accelerate disassembly of the long-lived intermediates revealed that Nup1p and Nup2p accelerate 16- and 19-fold the rate of dissociation of NLS cargo from Kap60p.Kap95p heterodimers; that Gsp1p-GTP accelerates > or = 447-fold the rate of dissociation of Kap60p.NLS-cargo from Kap95p; and that Nup2p and the Cse1p.Gsp1p-GTP complex independently accelerate > or = 22- and > or = 39-fold the rate of dissociation of NLS-cargo from Kap60p. We suggest that Nup1p, Nup2p, Cse1p, and Gsp1p accelerate disassembly of Kap95p.Kap60p.NLS-cargo complexes by triggering allosteric mechanisms within Kaps that cause rapid release of binding partners. In that way, Nup1p, Nup2p, Cse1p, and Gsp1p may function as karyopherin release factors (or KaRFs) in the nuclear basket structure of the S. cerevisiae NPC. PMID- 11867632 TI - Calcium-independent and cAMP-dependent modulation of soluble guanylyl cyclase activity by G protein-coupled receptors in pituitary cells. AB - It is well established that G protein-coupled receptors stimulate nitric oxide sensitive soluble guanylyl cyclase by increasing intracellular Ca(2+) and activating Ca(2+)-dependent nitric-oxide synthases. In pituitary cells receptors that stimulated adenylyl cyclase, growth hormone-releasing hormone, corticotropin releasing factor, and thyrotropin-releasing hormone also stimulated calcium signaling and increased cGMP levels, whereas receptors that inhibited adenylyl cyclase, endothelin-A, and dopamine-2 also inhibited spontaneous calcium transients and decreased cGMP levels. However, receptor-controlled up- and down regulation of cyclic nucleotide accumulation was not blocked by abolition of Ca(2+) signaling, suggesting that cAMP production affects cGMP accumulation. Agonist-induced cGMP accumulation was observed in cells incubated in the presence of various phosphodiesterase and soluble guanylyl cyclase inhibitors, confirming that G(s)-coupled receptors stimulated de novo cGMP production. Furthermore, cholera toxin (an activator of G(s)), forskolin (an activator of adenylyl cyclase), and 8-Br-cAMP (a permeable cAMP analog) mimicked the stimulatory action of G(s)-coupled receptors on cGMP production. Basal, agonist-, cholera toxin-, and forskolin-stimulated cGMP production, but not cAMP production, was significantly reduced in cells treated with H89, a protein kinase A inhibitor. These results indicate that coupling seven plasma membrane-domain receptors to an adenylyl cyclase signaling pathway provides an additional calcium-independent and cAMP-dependent mechanism for modulating soluble guanylyl cyclase activity in pituitary cells. PMID- 11867633 TI - Recognition of hybrid peptidyl carrier proteins/acyl carrier proteins in nonribosomal peptide synthetase modules by the 4'-phosphopantetheinyl transferases AcpS and Sfp. AB - The acyl carrier proteins (ACPs) of fatty acid synthase and polyketide synthase as well as peptidyl carrier proteins (PCPs) of nonribosomal peptide synthetases are modified by 4'-phosphopantetheinyl transferases from inactive apo-enzymes to their active holo forms by transferring the 4'-phosphopantetheinyl moiety of coenzyme A to a conserved serine residue of the carrier protein. 4' Phosphopantetheinyl transferases have been classified into two types; the AcpS type accepts ACPs of fatty acid synthase and some ACPs of type II polyketide synthase as substrates, whereas the Sfp type exhibits an extraordinarily broad substrate specificity. Based on the previously published co-crystal structure of Bacillus subtilis AcpS and ACP that provided detailed information about the interacting residues of the two proteins, we designed a novel hybrid PCP by replacing the Bacillus brevis TycC3-PCP helix 2 with the corresponding helix of B. subtilis ACP that contains the interacting residues. This was performed for the PCP domain as a single protein as well as for the TycA-PCP domain within the nonribosomal peptide synthetase module TycA from B. brevis. Both resulting proteins, designated hybrid PCP (hPCP) and hybrid TycA (hTycA), were modified in vivo during heterologous expression in Escherichia coli (hPCP, 51%; hTycA, 75%) and in vitro with AcpS as well as Sfp to 100%. The designated hTycA module contains two other domains: an adenylation domain (activating phenylalanine to Phe-AMP and afterward transferring the Phe to the PCP domain) and an epimerization domain (converting the PCP-bound l-Phe to d-Phe). We show here that the modified PCP domain of hTycA communicates with the adenylation domain and that the co-factor of holo-hPCP is loaded with Phe. However, communication between the hybrid PCP and the epimerization domain seems to be disabled. Nevertheless, hTycA is recognized by the next proline-activating elongation module TycB1 in vitro, and the dipeptide is formed and released as diketopiperazine. PMID- 11867634 TI - Identification of a familial hyperinsulinism-causing mutation in the sulfonylurea receptor 1 that prevents normal trafficking and function of KATP channels. AB - Mutations in the pancreatic ATP-sensitive potassium (K(ATP)) channel subunits sulfonylurea receptor 1 (SUR1) and the inwardly rectifying potassium channel Kir6.2 cause persistent hyperinsulinemic hypoglycemia of infancy. We have identified a SUR1 mutation, L1544P, in a patient with the disease. Channels formed by co-transfection of Kir6.2 and the mutant SUR1 in COS cells have reduced response to MgADP ( approximately 10% that of the wild-type channels) and reduced surface expression ( approximately 19% that of the wild-type channels). However, the steady-state level of the SUR1 protein is unaffected. Treating cells with lysosomal or proteasomal inhibitors did not improve surface expression of the mutant channels, suggesting that increased degradation of mutant channels by either pathway is unlikely to account for the reduced surface expression. Removal of the RKR endoplasmic reticulum retention/retrieval trafficking motif in either SUR1 or Kir6.2 increased the surface expression of the mutant channel by approximately 35 and approximately 20%, respectively. The simultaneous removal of the RKR motif in both channel subunits restored surface expression of the mutant channel to the wild-type channel levels. Thus, the L1544P mutation may interfere with normal trafficking of K(ATP) channels by causing improper shielding of the RKR endoplasmic reticulum retention/retrieval trafficking signals in the two channel subunits. PMID- 11867635 TI - Functional studies on recombinant domains of Mac-2-binding protein. AB - Mac-2-binding protein (M2BP) is a secreted glycoprotein suggested to have a role in host defense. It forms linear and ring-shaped oligomers, with each ring segment being composed of two monomers. We have produced recombinant human M2BP fragments comprising domains 1 and 2 (M2BP-1,2) and domains 3 and 4 (M2BP-3,4) in 293 human kidney cells to characterize structural and functional properties of M2BP. Both fragments were obtained in a native and glycosylated form, as analyzed by CD spectroscopy, trypsin susceptibility, and enzymatic deglycosylation. These results strongly suggest that both fragments are autonomous folding units. All three potential N-glycosylation sites in M2BP-1,2 and all four in M2BP-3,4 were found to be occupied. M2BP-1,2 expressed in tunicamycin-treated cells contained no glycosyl residues, indicating that O-glycosylation is not occurring. Ultracentrifugation revealed that M2BP-1,2 is homogeneously dimeric in the nanomolar range reflecting the properties of intact M2BP. Domain 2 (BTB/POZ domain) is thus identified as the dimerization domain of M2BP, because it has been formerly shown that recombinant domain 1 is monomeric. M2BP-3,4 showed a concentration-dependent self-association, and aggregates of different size and shape were shown by electron microscopy. In contrast to this irregular aggregation of M2BP-3,4, it has been formerly shown that a fragment comprising domains 2-4 still has the ability to form ring-like structures, although the rings are protein-filled, and thus domain 2 appears to be indispensable for ring formation. Solid phase assays showed that M2BP-3,4 contains binding sites for galectin-3, nidogen, and collagens V and VI, whereas M2BP-1,2 is inactive in binding. Both fragments showed no cell adhesive activity in contrast to native M2BP, suggesting that a concerted binding action and/or multivalent interactions of rings are necessary for cell attachment. PMID- 11867636 TI - The heme environment of recombinant human indoleamine 2,3-dioxygenase. Structural properties and substrate-ligand interactions. AB - Indoleamine 2,3-dioxygenase is a heme enzyme that catalyzes the oxidative degradation of L-Trp and other indoleamines. We have used resonance Raman spectroscopy to characterize the heme environment of purified recombinant human indoleamine 2,3-dioxygenase (hIDO). In the absence of L-Trp, the spectrum of the Fe(3+) form displayed six-coordinate, mixed high and low spin character. Addition of L-Trp triggered a transition to predominantly low spin with two Fe-OH(-) stretching modes identified at 546 and 496 cm(-1), suggesting H-bonding between the NH group of the pyrrole ring of L-Trp and heme-bound OH(-). The distal pocket of Fe(3+) hIDO was explored further by an exogenous heme ligand, CN(-); again, binding of L-Trp introduced strong H-bonding and/or steric interactions to the heme-bound CN(-). On the other hand, the spectrum of Fe(2+) hIDO revealed a five coordinate and high spin heme with or without L-Trp bound. The proximal Fe-His stretching mode, identified at 236 cm(-1), did not shift upon L-Trp addition, indicating that the proximal Fe-His bond strength is not affected by binding of the substrate. The high Fe-His stretching frequency suggests that Fe(2+) hIDO has a strong "peroxidase-like" Fe-His bond. Using CO as a structural probe for the distal environment of Fe(2+) hIDO revealed that binding of L-Trp in the distal pocket converted IDO to a peroxidase-like enzyme. Binding of L-Trp also caused conformational changes to the heme vinyl groups, which were independent of changes of the spin and coordination state of the heme iron. Together these data indicate that the strong proximal Fe-His bond and the strong H-bonding and/or steric interactions between l-Trp and dioxygen in the distal pocket are likely crucial for the enzymatic activity of hIDO. PMID- 11867637 TI - Fluoride exposure attenuates expression of Streptococcus pyogenes virulence factors. AB - Fluoridation causes an obvious reduction of dental caries by interference with cariogenic streptococci. However, the effect of fluoride on group A streptococci that causes rheumatic fever and acute poststreptococcal glomerulonephritis is not known. We have used proteomic analysis to create a reference proteome map for Streptococcus pyogenes and to determine fluoride-induced protein changes in the streptococci. Cellular and extracellular proteins were resolved by two dimensional polyacrylamide gel electrophoresis and identified by matrix-assisted laser desorption ionization mass spectrometry. 183 protein spots were visualized, and 74 spots representing 60 unique proteins were identified. A 16-h exposure to sodium fluoride caused decreased expression of proteins required to respond to cellular stress, including anti-oxidants, glycolytic enzymes, transcriptional and translational regulators, and protein folding. Fluoride caused decreased cellular expression of two well-characterized S. pyogenes virulence factors. Fluoride decreased expression of glyceraldehyde-3-phosphate dehydrogenase, which acts to bind fibronectin and promote bacterial adherence. We also performed proteomic analysis of protein released by S. pyogenes into the culture supernatant and observed decreased expression of M proteins following fluoride exposure. These data provide evidence that fluoride causes decreased expression by S. pyogenes proteins used to respond to stress, virulence factors, and implicated in non suppurative complications of S. pyogenes, including glomerulonephritis and rheumatic fever. PMID- 11867638 TI - E2Ls, E2F-like repressors of Arabidopsis that bind to E2F sites in a monomeric form. AB - E2F transcription factors are major regulators of cell proliferation, and each factor contributes differently to cell cycle control. Arabidopsis contains six E2F homologs, of which three are proteins that exhibit an overall similarity to animal E2Fs and interact with DPa and DPb to stimulate DNA binding. Here we describe the other three E2F-like proteins from Arabidopsis, E2L1-3, which have two copies of a domain with a limited similarity only to the DNA binding domain of E2F. Unlike known E2Fs, the three E2L proteins failed to interact with DPa and DPb and could efficiently bind E2F sites in a monomeric form through the dual type domain. Transfection assays revealed that E2Ls repress the transcription of reporter genes under the control of E2F-regulated promoters, indicating that E2Ls function to antagonize transactivation mediated by E2F.DP. When fused to green fluorescence protein, E2L1 and E2L3 were predominantly localized to the nucleus whereas E2L2 was detected in both the nucleus and cytoplasm. Because the transcripts of E2Ls were abundant in meristematic rather than fully differentiated tissues, E2Ls may balance the activities of E2F.DP and play a role in restraining cell proliferation. PMID- 11867639 TI - The Aes protein directly controls the activity of MalT, the central transcriptional activator of the Escherichia coli maltose regulon. AB - MalT, the transcriptional activator of the maltose regulon from Escherichia coli, is the prototype of a new family of transcription factors. Its activity is controlled by multiple regulatory signals. ATP and maltotriose (the inducer) are two effectors of the activator that positively control its multimerization, a critical step in promoter binding. In addition, MalK, the ABC component of the maltodextrin transport system, and the two enzymes MalY and Aes down-regulate MalT activity in vivo. By using a biochemical approach, we demonstrate here that (i) Aes controls MalT activity through direct protein-protein interaction, (ii) Aes competes with maltotriose for MalT binding, (iii) ATP and ADP differentially affect the competition between Aes and the inducer, and (iv) part, if not all, of the Aes binding site is located in DT1, the N-terminal domain of the activator, which also contains the ATP binding site. All of these characteristics point toward an identical mode of action for MalY and Aes. However, we have identified an amino acid substitution in MalT that suppresses MalT inhibition by Aes without interfering with its inhibition by MalY, suggesting that the binding sites of the two inhibitory proteins do not coincide. The differential effects of ATP and ADP on the competition between the inducer and Aes (or MalY) suggest that the ATPase activity displayed by MalT plays a role in the negative control of its activity. PMID- 11867640 TI - STO-609, a specific inhibitor of the Ca(2+)/calmodulin-dependent protein kinase kinase. AB - STO-609, a selective inhibitor of Ca(2+)/calmodulin-dependent protein kinase kinase (CaM-KK) was synthesized, and its inhibitory properties were investigated both in vitro and in vivo. STO-609 inhibits the activities of recombinant CaM-KK alpha and CaM-KK beta isoforms, with K(i) values of 80 and 15 ng/ml, respectively, and also inhibits their autophosphorylation activities. Comparison of the inhibitory potency of the compound against various protein kinases revealed that STO-609 is highly selective for CaM-KK without any significant effect on the downstream CaM kinases (CaM-KI and -IV), and the IC(50) value of the compound against CaM-KII is approximately 10 microg/ml. STO-609 inhibits constitutively active CaM-KK alpha (glutathione S-transferase (GST)-CaM-KK-(84 434)) as well as the wild-type enzyme. Kinetic analysis indicates that the compound is a competitive inhibitor of ATP. In transfected HeLa cells, STO-609 suppresses the Ca(2+)-induced activation of CaM-KIV in a dose-dependent manner. In agreement with this observation, the inhibitor significantly reduces the endogenous activity of CaM-KK in SH-SY5Y neuroblastoma cells at a concentration of 1 microg/ml (approximately 80% inhibitory rate). Taken together, these results indicate that STO-609 is a selective and cell-permeable inhibitor of CaM-KK and that it may be a useful tool for evaluating the physiological significance of the CaM-KK-mediated pathway in vivo as well as in vitro. PMID- 11867641 TI - Heterogeneous nuclear ribonucleoprotein (hnRNP) K is a component of an intronic splicing enhancer complex that activates the splicing of the alternative exon 6A from chicken beta-tropomyosin pre-mRNA. AB - Splicing of the chicken beta-tropomyosin exon 6A is stimulated, both in vivo and in vitro, by an intronic pyrimidine-rich element (S4) located 37 nucleotides downstream of exon 6A. Several pyrimidine-rich sequences are able to substitute for the natural S4 enhancer with various stimulatory effects. We show that the different enhancer sequences recruit U1 small nuclear ribonucleoprotein (SnRNP) to the exon 6A 5' splice site, with an efficiency that correlates with the splicing activation. By using RNA affinity and two-dimensional gel electrophoresis, we characterized several proteins that bind to the different enhancer sequences. Heterogeneous nuclear ribonucleoprotein (hnRNP) K and hnRNP I (polypyrimidine track-binding protein, PTB) exhibit a higher level of interaction with the strong enhancer sequences (S4) than with the weakest enhancers. Functional analysis shows that hnRNP K is a component of the enhancer complex that promotes exon 6A splicing through the wild-type S4 sequence. The addition of recombinant hnRNP K to nuclear extracts preincubated with poly(rC) RNA competitor completely restores splicing efficiency to the original level. hnRNP I (PTB) was also found associated with the strong enhancer sequences. Its function in the splicing of exon 6A is discussed. PMID- 11867642 TI - Proteins from Mucuna pruriens and enzymes from Echis carinatus venom: characterization and cross-reactions. AB - Mucuna pruriens seeds have been widely used against snakebite in traditional medicine. The antivenin property of a water extract of seeds was assessed in vivo in mice. The serum of mice treated with extract was tested for its immunological properties. Two proteins of Echis carinatus venom with apparent molecular masses of 25 and 16 kDa were detected by Western blot analysis carried out using IgG of mice immunized with extract or its partially purified protein fractions. By enzymatic in-gel digestion and electrospray ionization-mass spectrometry/mass spectrometry analysis of immunoreactive venom proteins, phospholipase A(2,) the most toxic enzyme of snake venom, was identified. These results demonstrate that the observed antivenin activity has an immune mechanism. Antibodies of mice treated with non-lethal doses of venom reacted against some proteins of M. pruriens extract. Proteins of E. carinatus venom and M. pruriens extract have at least one epitope in common as confirmed by immunodiffusion assay. PMID- 11867643 TI - The low density lipoprotein receptor-related protein mediates fibronectin catabolism and inhibits fibronectin accumulation on cell surfaces. AB - Low density lipoprotein receptor-related protein (LRP) is a member of the low density lipoprotein receptor family, which functions as an endocytic receptor for diverse ligands. In this study, we demonstrate that murine embryonic fibroblasts (MEF-2 cells) and 13-5-1 Chinese hamster ovary cells, which are LRP-deficient, accumulate greatly increased levels of cell-surface fibronectin (Fn), compared with LRP-expressing MEF-1 and CHO-K1 cells. Increased Fn was also detected in conditioned medium from LRP-deficient MEF-2 cells; however, biosynthesis of Fn by MEF-1 and MEF-2 cells was not significantly different. When LRP-deficient cells were dissociated from monolayer culture, increased levels of Fn remained with the cells, as determined by cell-surface protein biotinylation, suggesting an intimate relationship with cell surface-binding sites. The LRP antagonist, receptor-associated protein (RAP), promoted Fn accumulation in association with MEF-1 cells, whereas expression of full-length LRP in MEF-2 cells substantially decreased Fn accumulation, confirming the role of LRP in this process. Purified LRP bound directly to immobilized Fn, and this interaction was inhibited by RAP. Furthermore, MEF-1 cells degraded (125)I-Fn at an increased rate, compared with MEF-2 cells. (125)I-Fn degradation by MEF-1 cells was inhibited by RAP. These results demonstrate that LRP functions as a catabolic receptor for Fn. The function of LRP in Fn degradation and the ability of LRP to regulate levels of other plasma membrane proteins represent possible mechanisms whereby LRP prevents Fn accumulation on cell surfaces. PMID- 11867644 TI - Heat shock changes the heterogeneity distribution in populations of Caenorhabditis elegans: does it tell us anything about the biological mechanism of stress response? AB - In this paper we analyze survival data of populations of sterilized nematodes, Caenorhabditis elegans, exposed to heat shocks of different duration at the beginning of their adult lives. There are clear hormesis effects after short exposure to heat and clear debilitation effects after long exposure. Intermediate durations result in a mixture of these two effects. In this latter case, the survival curves for the control and experimental populations intersect. We show that observed effects may be explained by using a model of discrete heterogeneity. According to this model, each population of worms in the experiment is a mixture of subcohorts of frail, normal, and robust individuals; exposure to heat changes the initial proportion of worms in the subcohorts (heterogeneity distribution); and these changes depend on the duration of exposure. In other words, exposure to heat does not influence mortality rates (survival functions) in the subcohorts but does cause individuals to move from one subcohort to another. In a biological interpretation of this finding we hypothesize that, when coping with stress, the organisms of worms use several lines of defense. Switching these lines on and off in response to stress in individual organisms generates the spectrum of observed survival effects at the population level. We discuss possible molecular biological mechanisms of stress response and directions for further research. PMID- 11867645 TI - Histone acetyltransferase activities of cAMP-regulated enhancer-binding protein and p300 in tissues of fetal, young, and old mice. AB - CBP, a protein that binds to cyclic adenosine monophosphate-regulated enhancer binding protein, and homologue protein, p300, have histone acetyltransferase (HAT) activity and are important in gene transcription, although their physiological functions in vivo remain to be further elucidated. By using immunoprecipitation and HAT activity assay we have found that p300 and CBP have similar tissue patterns of HAT activities, with the highest level in the brain, a relatively high level in the lung, spleen, and heart, an intermediate level in testes and muscle, and a lower level in liver and kidney; that HAT activities of p300 and CBP are relatively stable with advancing age in most examined tissues, but in liver, muscle, and testes, the activities are attenuated with aging; and that HAT activities of p300 and CBP are high in the brain and liver of E14 fetal and newborn mice. These data suggest that the HAT activities of p300 and CBP are important for gene transcription involved in tissue-specific expression, aging, and developing processes. PMID- 11867646 TI - Gene expression profile of long-lived snell dwarf mice. AB - To gain further insight into the basis for the extended longevity and delayed aging of Snell dwarf (dw/dw) mice, we have measured levels of expression of 2352 genes in liver of mice at 6 months of age. We find 60 genes for the which the Student's t statistic meets the arbitrary criterion of p <.001, and among these 17 meet the Bonferroni-adjusted significance criterion at p <.05, which corresponds to a nominal value of p <.00002. Using the Bonferroni criterion, we find that dwarf mice show increases in liver mRNA for two mannose-binding lectins, two DNA binding proteins, serum amyloid P component, corticosteroid binding globulin, and insulin-like growth factor-binding protein 2, as well as decreases in a two phosphodiesterases, a pheromone-binding urinary protein, insulin-like growth factor-I (IGF-I), a calcium-binding protein calgranulin B, a deubiquitinating enzyme, a hydroxysteroid dehydrogenase, a DNA methyltransferase, a glycine transporter, and a placental lactogen. We also use this data set to compare the results of different suggested criteria for evaluating intergroup differences in gene expression. Of the 2352 genes examined, 524 (22%) showed a twofold difference between dwarf and normal mice, but most of these fail to meet the conventional significance criterion of p <.05, let alone criteria that have been adjusted to compensate for multiple comparison artifacts. The list of genes that show reliable differences between dwarf and control animals provides new insights into the range of changes induced by deficiencies in growth hormone, thyroid-stimulating hormone, and prolactin, and it will help to guide further studies of the pathways by which these hormone deficiencies contribute to delayed aging in these mutant mice. PMID- 11867647 TI - Multiple stressors in Caenorhabditis elegans induce stress hormesis and extended longevity. AB - We demonstrate here that the nematode Caenorhabditis elegans displays broad hormetic abilities. Hormesis is the induction of beneficial effects by exposure to low doses of otherwise harmful chemical or physical agents. Heat as well as pretreatment with hyperbaric oxygen or juglone (a chemical that generates reactive oxygen species) significantly increased subsequent resistance to the same challenge. Cross-tolerance between juglone and oxygen was also observed. The same heat or oxygen pretreatment regimens that induced subsequent stress resistance also increased life expectancy and maximum life span of populations undergoing normal aging. Pretreatment with ultraviolet or ionizing radiation did not promote subsequent resistance or increased longevity. In dose-response studies, induced thermotolerance paralleled the induced increase in life expectancy, which is consistent with a common origin. PMID- 11867648 TI - Dynamics of stability: the physiologic basis of functional health and frailty. AB - Under basal resting conditions most healthy physiologic systems demonstrate highly irregular, complex dynamics that represent interacting regulatory processes operating over multiple time scales. These processes prime the organism for an adaptive response, making it ready and able to react to sudden physiologic stresses. When the organism is perturbed or deviates from a given set of boundary conditions, most physiologic systems evoke closed-loop responses that operate over relatively short periods of time to restore the organism to equilibrium. This transiently alters the dynamics to a less complex, dominant response mode, which is denoted "reactive tuning." Aging and disease are associated with a loss of complexity in resting dynamics and maladaptive responses to perturbations. These alterations in the dynamics of physiologic systems lead to functional decline and frailty. Nonlinear mathematical techniques that quantify physiologic dynamics may predict the onset of frailty, and interventions aimed toward restoring healthy dynamics may prevent functional decline. PMID- 11867649 TI - The prevalence of depressive symptoms. PMID- 11867650 TI - What was the disease of the bones that affected King David? AB - BACKGROUND: Elderly people have suffered from pain in their bones, which may be associated with various diseases, for thousands of years. METHODS: This report analyzes the disease that affected the Biblical King David, the second and greatest of Israel's Kings, who ruled the country 3000 years ago. RESULTS: The sentences "My strength failed.and my bones are consumed," and "My bones wasted away through my anguished roaring all day long" indicate that King David suffered from osteoporosis, which affected his bones. Among the various diseases that may be associated with osteoporosis, the most likely are senile osteoporosis, hyperparathyroidism, or malignant disease. Among these diseases, the diagnosis of malignancy is the most acceptable. CONCLUSION: This report demonstrates that the roots of contemporary modern gerontology can be traced to Biblical times. PMID- 11867651 TI - Prevalence rate and correlates of depressive symptoms in older individuals: the Veneto Study. AB - BACKGROUND: Depressive symptoms (DS) are very common in elders, particularly among women, and their prevalence rates vary widely across countries. METHODS: A random sample of noninstitutionalized men (n = 867) and women (n = 1531), aged 65 years and older, from the Veneto region, northeastern Italy, were interviewed and participated in a brief physical examination in their homes. The prevalence rates of DS and the associated physical, social, and psychological factors were analyzed. RESULTS: The overall prevalence rate of DS was 58% in women and 34% in men (p <.0001), but there was no significant trend with age. Women were at higher risk of DS (OR = 1.63) than men, even after adjusting for traditional risk factors, such as fair-poor self-rated health, sleep disturbances and use of sleep medications, lack of support from social and family network, and physical and cognitive impairment. CONCLUSIONS: This study provides evidence that older women more frequently report DS than men, independently from the presence of traditional risk and associated factors. Cultural and lifestyle factors throughout life might explain this gender difference. PMID- 11867652 TI - Looking at the future of geriatric care in developing countries. AB - During the next 20 years, many less developed countries (LDC) will have age structures approaching those of the present time in more developed countries (MDC). This is occurring more rapidly in the LDC of Asia and Latin America. The future of aging populations in LDC is dependent on the degree of poverty in these countries. Poverty is a major determinant of disability and mortality in older persons. With the march of globalization, diseases in LDC are changing from infectious to noncommunicable diseases, such as diabetes. Nevertheless, infections such as tuberculosis still take a major toll on the elderly. The epidemiological transition in LDC has created a need for health care transitions from systems based on cure to ones that highlight prevention and long-term care. LDC have the opportunity to develop systems that differ from those in MDC by capitalizing on the lack of infrastructure to produce more home-based rather than institution-based long-term care systems. Involvement of the elderly in the planning of their own futures is of paramount importance. Appropriate planning now will decide the future of the elderly in LDC during the next 20 to 40 years. PMID- 11867653 TI - Anaerobic power and physical function in strength-trained and non-strength trained older adults. AB - BACKGROUND: Challenging daily tasks, such as transferring heavy items or rising from the floor, may be dependent on the ability to generate short bursts of energy anaerobically. The purposes of this study were to determine if strength trained (ST) older adults have higher anaerobic power output compared with non strength-trained (NST) older adults and to determine the relationship between anaerobic power and performance-based physical function. METHODS: Thirty-five men and women (age 71.5 +/- 6.4 years, mean +/- SD; NST: n = 18, ST: n = 17) were grouped by training status. Outcome variables included relative anaerobic power (Wingate test), physical function measured with the Continuous Scale Physical Functional Performance Test (CS-PFP, scaled 0 to 100), and anthropometric lean thigh volume (LTV). Analysis of covariance (with age and sex as covariates) was used to determine group differences in the dependent variables listed above. Pearson's r was used to determine the relationship between anaerobic power, CS PFP total score (TOT), and CS-PFP lower body strength domain score (LBS). RESULTS: The ST group had significantly higher mean anaerobic power (NST 58.9 +/- 16 W/l, ST 96.3 +/- 23 W/l), CS-PFP total (NST 61.2 +/- 13, ST 73.7 +/- 8), and LBS (NST 54.1 +/- 17, ST 70.9 +/- 8) compared with the NST group (p <.05). However, LTV was similar for both groups (NST 3.323 +/- 0.75; ST 3.179 +/- 0.79), which suggests that the ST group had higher muscle quality compared with the NST group. Anaerobic power was significantly related to TOT (r =.611, p =.001) and LBS (r =.650, p =.001). CONCLUSIONS: High levels of physical function in ST older adults may in part be explained by higher levels of anaerobic power associated with strength training. PMID- 11867654 TI - Prevalence and impact of medical comorbidity in Alzheimer's disease. AB - BACKGROUND: We examined the prevalence of comorbid medical illnesses in Alzheimer's disease (AD) patients at different severity levels. We also examined the effect of cumulative medical comorbidity on cognition and function. METHODS: Analyses of data from 679 AD patients (Mini-Mental State Exam score range 0-30, mean +/- SD = 11.8 +/- 8) from 13 sites (four dementia centers assessing outpatients, four managed care organizations, two assisted living facilities, and three nursing homes) prospectively recruited using a stratification approach including dementia severity and care setting. Medical comorbidity was quantified using the Cumulative Illness Rating Scale-Geriatric. RESULTS: Across patients, 61% had three or more comorbid medical illnesses. Adjusting for age, gender, race, and care setting, medical comorbidity increased with dementia severity (mild to moderate, p <.01; moderate to severe, p <.001). Adjusting for age, educational level, gender, race, and care setting, higher medical comorbidity was associated with greater impairment in cognition (p <.001) and in self-care (p <.001). CONCLUSIONS: Despite the limitation of a cross-sectional design, our initial findings suggest that there is a strong association between medical comorbidity and cognitive status in AD. Optimal management of medical illnesses may offer potential to improve cognition in AD. PMID- 11867655 TI - Prevalence and association of ventricular tachycardia and complex ventricular arrhythmias with new coronary events in older men and women with and without cardiovascular disease. AB - BACKGROUND: We report the prevalence of ventricular tachycardia (VT) and of complex ventricular arrhythmias (VA) and their association with new coronary events in older men and women. METHODS: The prevalence of VT and of complex VA detected by 24-hour ambulatory electrocardiograms and the incidence of new coronary events in older persons with coronary artery disease (CAD), with hypertension, valvular disease, or cardiomyopathy without CAD, and with no cardiovascular disease was investigated in 915 men (mean age 80 +/- 8 years) and in 1,874 women (mean age 81 +/- 8 years) in a long-term health care facility. Follow-up was 45 +/- 30 months in men and 47 +/- 30 months in women. RESULTS: The prevalence of VT was 16% in men and 15% in women with CAD, 9% in men and 8% in women with hypertension, valvular disease, or cardiomyopathy without CAD, and 3% in men and 2% in women with no cardiovascular disease. The prevalence of complex VA was 69% in men and 68% in women with CAD, 54% in men and 55% in women with hypertension, valvular disease, or cardiomyopathy without CAD, and 31% in men and 30% in women with no cardiovascular disease. In men and in women with CAD or with hypertension, valvular disease, or cardiomyopathy, VT and complex VA increased the incidence of new coronary events (p <.0001). Within each of the groups of patients, the incidences of new coronary events in men and in women with and without VT or complex VA were similar. CONCLUSIONS: The prevalence of VT and of complex VA were similar in older men and women. VT and complex VA were associated with a higher incidence of new coronary events in men and women with CAD or with hypertension, valvular disease, or cardiomyopathy without CAD, but not in men and women with no cardiovascular disease. PMID- 11867656 TI - Predictors of decline in MMSE scores among older Mexican Americans. AB - BACKGROUND: The purpose of this analysis was to examine the association of sociodemographic variables and health-related conditions with 5-year declines in cognitive function among Mexican American elderly persons. METHODS: The cognitive function of 1759 participants was assessed by using the Mini-Mental State Examination (MMSE) in 1993/1994 and again in 1998/1999. Cognitive decline was defined by two sets of criteria: (1) a drop to 17 or below on the MMSE at follow up, and (2) a decline of at least three points, the mean change in MMSE scores among respondents who obtained scores at or above the 5th percentile distribution at baseline. RESULTS: Cognitive decline was significantly associated with sociodemographic variables including age, education, marital status, and household composition. In addition, respondents with reported vision impairment, stroke, and diabetes were at increased risk for cognitive decline after controlling for multiple potential confounders. CONCLUSION: Although age and education have been reported as the more salient predictors of cognitive deterioration, other sociodemographic and several medical conditions including stroke and diabetes should be considered as part of cognitive aging studies among Mexican American elders. PMID- 11867658 TI - Variability in reaction time performance of younger and older adults. AB - Age differences in three basic types of variability were examined: variability between persons (diversity), variability within persons across tasks (dispersion), and variability within persons across time (inconsistency). Measures of variability were based on latency performance from four measures of reaction time (RT) performed by a total of 99 younger adults (ages 17--36 years) and 763 older adults (ages 54--94 years). Results indicated that all three types of variability were greater in older compared with younger participants even when group differences in speed were statistically controlled. Quantile-quantile plots showed age and task differences in the shape of the inconsistency distributions. Measures of within-person variability (dispersion and inconsistency) were positively correlated. Individual differences in RT inconsistency correlated negatively with level of performance on measures of perceptual speed, working memory, episodic memory, and crystallized abilities. Partial set correlation analyses indicated that inconsistency predicted cognitive performance independent of level of performance. The results indicate that variability of performance is an important indicator of cognitive functioning and aging. PMID- 11867657 TI - Risk factors for motor vehicle crashes in older women. AB - BACKGROUND: Motor vehicle crash and fatality rates are higher per mile driven for elderly drivers, with an exponential increase above age 75. Identifying elderly drivers who are at risk for automobile crashes may help direct interventions to reduce their high rate of injuries and deaths. METHODS: Subjects were 1416 women aged 65 to 84 enrolled in the Portland, Ore. site of the Study of Osteoporotic Fractures. Motor vehicle crash information for the years 1986-1995 for each participant was obtained from the Oregon State Department of Transportation. Items from questionnaires, interviews, and physical examinations were tested prospectively for associations with the occurrence of motor vehicle crashes. RESULTS: About one third of participants (415 of 1416) had a motor vehicle crash during a mean follow-up time of 5.7 years. After adjustment for age and weekly driving mileage, risk factors significantly associated with motor vehicle crashes were a fall in the previous year [hazard ratio (HR) 1.53, 95% confidence interval (CI) 1.26-1.86], a greater orthostatic systolic blood pressure drop (HR 1.11 per 12.5 mm Hg, 95% CI 1.01-1.22), and increased foot reaction time (HR 1.10 per 0.06 second, 95% CI 1.00-1.22). Other neuromuscular tests, functional status, medical diagnoses, vision tests, and cognitive tests did not predict motor vehicle crashes in this study population. CONCLUSIONS: This prospective study with extended follow-up of a large cohort of elderly women has identified crash risk factors that can be measured in the clinical setting. Further study is needed to determine if interventions aimed at these risk factors can decrease the risk of motor vehicle crashes. PMID- 11867659 TI - Personality traits and existential concerns as predictors of the functions of reminiscence in older adults. AB - This study examines to what extent personality and existential constructs predict the frequency of reminiscence, in general, and its various functions, in particular. Eighty-nine older adults completed the NEO-Five Factor Inventory, the Life Attitude Profile--Revised, and the Reminiscence Functions Scale. Neuroticism predicted total reminiscence frequency, as well as reminiscence for self understanding and ruminating about a negative past. Extraversion predicted total reminiscence frequency, as well as reminiscence for generating stimulation, conversation, and maintaining memories of departed loved ones. Openness to experience predicted total reminiscence frequency and reminiscence for addressing life meaning and death. Existential concerns, and in particular low desire to seek new challenges, added significant additional predictive power for total reminiscence frequency and for such uses as generating stimulation, preparing for death, and ruminating about the past. The discussion draws the implications of the finding that the combination of personality traits and existential concerns predicted the overall reminiscence frequency together with the intrapersonal functions of reminiscence. PMID- 11867661 TI - Evidence for genetic mediation of executive control: a study of aging male twins. AB - The objective of this study was to investigate the genetic and environmental influences on indexes of executive control in elderly male twins (members of the National Heart, Lung, and Blood Institute Twin Study). Multivariate genetic modeling was applied to performance on four tests: Digit Symbol Substitution, color--word interference, Trail Making B, and verbal fluency. After exclusion of individuals with a positive stroke history, data were available for 80 monozygotic and 78 dizygotic twin pairs of ages 69--80 years. Performance on all measures was adjusted for age and education. Significant genetic and environmental influences to performance on each measure of executive control were identified (range of heritability = 34%--68%). Multivariate analyses revealed that a model with a latent executive control factor most adequately fit the observed covariances on test performance, chi(2)(58, N = 316) = 69.7, p =.14. The shared executive control factor had a heritability of 79% and accounted for 10%- 56% of the genetic variance in performance on each of the four tests. Of the 4 tests examined in this analysis, Digit Symbol Substitution appeared to be the marker of executive control with the largest genetic component, whereas verbal fluency stood out as displaying a pattern of genetic and environmental influences distinct from the other 3 measures. PMID- 11867660 TI - Exercise and depressive symptoms: a comparison of aerobic and resistance exercise effects on emotional and physical function in older persons with high and low depressive symptomatology. AB - This study examines and compares the effect of aerobic and resistance exercise on emotional and physical function among older persons with initially high or low depressive symptomatology. Data are from the Fitness, Arthritis and Seniors Trial, a trial among 439 persons 60 years or older with knee osteoarthritis randomized to health education (control), resistance exercise, or aerobic exercise groups. Depressive symptoms (assessed by the Center for Epidemiologic Studies--Depression scale) and physical function (disability, walking speed, and pain) were assessed at baseline and after 3, 9, and 18 months. Compared with results for the control group, aerobic exercise significantly lowered depressive symptoms over time. No such effect was observed for resistance exercise. The reduction in depressive symptoms with aerobic exercise was found both among the 98 participants with initially high depressive symptomatology and among the 340 participants with initially low depressive symptomatology and was the strongest for the most compliant persons. Aerobic and resistance exercise significantly reduced disability and pain and increased walking speed both, and to an equal extent, in persons with high depressive symptomatology and persons with low depressive symptomatology. PMID- 11867662 TI - Age changes and differences in personality traits and states of the old and very old. AB - The purpose of this study was to examine stability and change in personality traits and states for 3 age groups: centenarians, octogenarians, and sexagenarians. One hundred seventy-nine older adults participated in the 2-wave study. Results concerning age-group differences indicated that centenarians scored higher in Suspiciousness but lower in Intelligence and Stress when compared with the other 2 age groups. Octogenarians were lower in the personality traits Intelligence, Dominance, and Conscientiousness when compared with sexagenarians. Octogenarians were lower in the personality state Arousal, but higher on Regression. Results from the longitudinal analyses for centenarians indicated lower scores for Sensitivity, but higher scores for Radicalism (both personality traits), as well as higher scores of Fatigue and Depression (personality states) at follow-up. For the younger 2 age groups, age changes included higher scores for Sensitivity and Suspiciousness (personality traits). Stability scores for traits and states were considerably lower for centenarians when compared with the younger age groups. PMID- 11867663 TI - Activity in older adults: cause or consequence of cognitive functioning? A longitudinal study on everyday activities and cognitive performance in older adults. AB - The impact of three types of everyday activities (i.e., social, experiential, and developmental) on four cognitive functions (i.e., immediate recall, learning, fluid intelligence, and information-processing speed) and one global indicator of cognitive functioning (Mini-Mental State Exam score) over a period of 6 years was studied in a large 55--85 year-old population-based sample (N = 2,076). A cross lagged regression model with latent variables was applied to each combination of 1 cognitive function and 1 type of activity, resulting in 15 (3 x 5) different models. None of the activities were found to enhance cognitive functioning 6 years later when controlling for age, gender, level of education, and health, as well as for unknown confounding variables. Conversely, one cognitive function (i.e., information-processing speed) appeared to affect developmental activity. It is suggested that no specific activity, but rather socioeconomic status to which activities are closely connected, contributes to maintenance of cognitive functions. PMID- 11867664 TI - Education, wealth, and cognitive function in later life. AB - Population-based studies of health often use education as the sole indicator of socioeconomic status (SES); the independent contributions of education and other SES covariates are rarely delineated. Using Wave 1 of the Asset and Health Dynamics Among the Oldest Old study, the authors examined the extent to which educational attainment influences performance on three separate domains of cognitive status by race and Latino ethnicity and introduced controls for wealth and household income. Results indicate that the education effect is minimally weakened after adjusting for wealth; the wealth effect, however, is greatly attenuated after adjusting for education. Blacks and Whites exhibited a similar education--cognition relationship; Latino elderly did not experience commensurate gains in cognitive function with increasing education. Results suggest that although the education--cognition relationship may in part reflect an SES gradient, the association is more likely due to the process and consequences of education itself. PMID- 11867665 TI - Differential age effects in semantic and episodic memory. AB - Results from 4 experimental tasks and 8 data sets (the 4 tasks involved either multiple sessions or different stimuli) as well as a vocabulary test conducted on the same 80 participants (40 younger and 40 older adults) are reported. The authors employed 2 semantic memory tasks (lexical decision and multiplication verification) using data from 2 sessions (for a total of 4 semantic data sets) and 2 episodic memory tasks (hybrid visual search and memory search with digits and with words as stimuli). Factor analyses using slope and intercept data from the 8 experimental data sets indicated the presence of 3 latent factors: a single intercept factor for both episodic and semantic tasks and separate slope factors for episodic and semantic tasks. A structural equation model with paths from age to 3 different 1st-order latent factors (episodic central processes, semantic central processes, and combined episodic and semantic peripheral processes) fit better than general factor models. These data are consistent with a theoretical framework in which there are age-related dissociations between peripheral and central processes across semantic and episodic memory. PMID- 11867666 TI - Usefulness of a 15-item version of the Boston Naming Test in neuropsychological assessment of low-educational elders with dementia. AB - The purpose of this study was to validate a reduced version (15 items) of the Boston Naming Test (BNT) in a sample of 78 low-educational elderly persons with or without dementia, as determined by independent assessment with a battery of cognitive tests. The reduced version was found to be equivalent to the complete BNT, and to have criterion validity with respect to other measures of dementia. We conclude that the reduced version is a useful instrument for assessing patients who require shorter testing methods because of severe cognitive deterioration or their low level of education. PMID- 11867668 TI - The Therapeutic Environment Screening Survey for Nursing Homes (TESS-NH): an observational instrument for assessing the physical environment of institutional settings for persons with dementia. AB - OBJECTIVE: To develop an observational instrument that describes the ability of physical environments of institutional settings to address therapeutic goals for persons with dementia. METHODS: A National Institute on Aging workgroup identified and subsequently revised items that evaluated exit control, maintenance, cleanliness, safety, orientation/cueing, privacy, unit autonomy, outdoor access, lighting, noise, visual/tactile stimulation, space/seating, and familiarity/homelikeness. The final instrument contains 84 discrete items and one global rating. A summary scale, the Special Care Unit Environmental Quality Scale (SCUEQS), consists of 18 items. Lighting items were validated using portable light meters. Concurrent criterion validation compared SCUEQS scores with the Professional Environmental Assessment Protocol (PEAP). RESULTS: Interrater kappa statistics for 74% of items were above.60. For another 10% of items, kappas could not be calculated due to empty cells, but interrater agreement was above 80%. The SCUEQS demonstrated an interrater reliability of.93, a test--retest reliability of.88, and an internal consistency of.81--.83. Light meter ratings correlated significantly with the Therapeutic Environment Screening Survey for Nursing Homes (TESS-NH) lighting items (r =.29--.38, p =.01--.04), and the SCUEQS correlated significantly with global PEAP ratings (r =.52, p <.01). DISCUSSION: The TESS-NH efficiently assesses discrete elements of the physical environment and has strong reliability and validity. The SCUEQS provides a quantitative measure of environmental quality in institutional settings. PMID- 11867669 TI - Dying away from home: quandaries of migration for elders in two ethnic groups. AB - OBJECTIVES: The purpose was to examine how Cambodian Americans and Filipino Americans view their homeland in old age and how those views affect the contemplation of death, using a multifaceted theoretical framework that encompasses transnationality, place, ethnic identity, continuity, and cultural phenomenology. METHODS: Three in-depth interviews over a 1-year period were conducted with 126 respondents (48 Cambodian Americans and 78 Filipino Americans). Both open-ended and semistructured questions were asked to determine how respondents viewed their eventual deaths. RESULTS: Many Cambodian Americans and Filipino Americans expressed a desire to die in their homelands. Whether or not they desired to return to the homeland to die was mediated by the presence or absence of the extended family, memories of the homeland, and the availability of traditional ritual practices in the United States. DISCUSSION: The preoccupation of respondents with where to die apparently reflects the desire to create continuity in their lives. In their efforts to reconcile issues of continuity, cultural meanings surrounding memory, ritual, and the family were paramount and appeared to reflect a desire to bring closure to unresolved conflicts in their lifetime. This research, including its theoretical framework, has implications for using the concept of transnationality and related concepts in gerontological research. PMID- 11867670 TI - Religion, death of a loved one, and hypertension among older adults in Japan. AB - OBJECTIVE: To see whether three dimensions of religion (private religious practices, religious coping, and belief in the afterlife) buffer the effect of the death of a significant other on change in self-reported hypertension over time. METHODS: Interviews were conducted with a nationally representative sample of people aged 60 and older in Japan at two points in time, 1996 and 1999. Complete data were available on 1,723 older Japanese. Respondents were asked a series of questions about their religious beliefs and practices, whether a family member or close friend had died in the past year, and whether they had hypertension. RESULTS: The data suggest that older adults in Japan who experienced the death of a loved one but who believed in a good afterlife were less likely to report they had hypertension at the follow-up interview than elderly people in Japan who lost a close other but did not believe in a good afterlife. DISCUSSION: The results suggest how one overlooked dimension of religion (i.e., religious beliefs) may bolster the health of older people in the face of adversity. PMID- 11867671 TI - Life course effects of early parental loss among very old African Americans. AB - OBJECTIVE: To analyze the life course effects of the early loss of one or both parents on very old Black Americans. METHODS: Open-ended, semistructured interviews were used with a sample of 109 respondents aged 85 years and older. Correlations identified significant associations, and qualitative data illustrate life course trajectories of selected respondents. RESULTS: Those who lost a parent through death or desertion were less integrated into family and friendship groups in late life, and they had fewer social resources in general. Qualitative data describe three outcomes in the sample: those who grew up with both parents present, those who lost a parent but still reported a contented childhood, and those with disrupted families and negative effects. DISCUSSION: The respondents' open-ended commentary about their past lives and their current situation enhances understanding of connections between early life events and adaptation in old age. PMID- 11867672 TI - Interlocking trajectories of loss-related events and depressive symptoms among elders. AB - OBJECTIVES: As people age, their peers (who are also aging) become increasingly susceptible to health decline and death, implying potential growth in stressful loss-related events over time for the individual. Yet little research has examined trajectories of stress and their relationship to trajectories of depression among elders. The purpose of this research was to determine whether growth in loss-related events occurs for elders and whether stress growth is related to the well-known growth in depressive symptomatology in later life. METHODS: Three waves of National Institute on Aging Established Populations for Epidemiologic Studies of the Elderly (Duke University site) data were used in the analyses. Latent growth curve models were estimated for stress, for depressive symptoms, and for stress predicting depression net of several covariates. RESULTS: Findings include that (a) loss-events evidence clear growth across age at the aggregate level, but with much variation within the sample, and (b) variation in growth in stress is strongly related to variation in growth in depressive symptoms. DISCUSSION: The results suggest that stress in later life may be conceived of as a growth process, with strong consequences for trajectories of mental health. PMID- 11867673 TI - Another look at aggregate changes in severe cognitive impairment: further investigation into the cumulative effects of three survey design issues. AB - OBJECTIVES: This study explored whether previously reported declines in severe cognitive impairment were robust to cumulative effects of potentially confounding survey design issues. METHODS: Using the 1993 Asset and Health Dynamics of the Oldest Old study (n = 7,443) and 1998 Health and Retirement Survey (HRS; n = 7,624) the proportion of persons ages 70 and older with severe cognitive impairment was calculated under various assumptions about item nonresponse, differential loss to follow-up, and the size and composition of the nursing home population. Impairment was measured for self-respondents using a modified version of the Telephone Interview Cognitive Screen; for proxy respondents ratings of memory and judgment were used. Chi-square tests were adjusted to account for complex survey designs. RESULTS: Ignoring loss to follow-up, excluding nursing home residents, and assigning a low score to those refusing subscales yielded a statistically significant decline in severe cognitive impairment from 5.8% in 1993 to 3.8% in 1998, or an average annual decline of 6.9%. When cumulative effects of survey design issues were considered and design effects incorporated into statistical tests, statistically significant declines persisted, albeit at a reduced average annual rate, ranging from 2.5% to 6.9% per year. DISCUSSION: Previously reported improvements in severe cognitive impairment appear to be robust to a variety of specifications. Replication with future waves of the HRS and other data is warranted. PMID- 11867675 TI - The role of growth hormone in T-cell development and reconstitution. AB - Growth hormone (GH), directly or through GH-induction of insulin-like growth factor (IGF)-1, has been implicated in lymphocyte development and function. Recent studies have questioned the role of GH and IGF-1 in immune responses. This review examines experimental data describing the immunoregulatory function of GH and attempts to reconcile the literature. PMID- 11867676 TI - The role of PPARs in inflammation and immunity. AB - The family of transcription factors termed peroxisome proliferator-activated receptors (PPARs) has recently been the focus of much interest for their possible role in the regulation of inflammation and immune responses. PPARalpha and PPARgamma have been implicated in the regulation of macrophage and endothelial cell inflammatory responses. Although PPAR activation has generally been shown to have anti-inflammatory effects, opposite effects have been noted, and results often appear to depend on the ligands being used and the inflammatory parameters being measured. Recently, my laboratory and others have described a role for PPARgamma in the responses of T lymphocytes. Ligands for PPARgamma have been found to inhibit proliferation of activated T cells, and this appears to involve inhibition of IL-2 secretion and/or the induction of apoptosis. However, one problem in the interpretation of many of the studies of PPARgamma, inflammation, and immunity is that ligands thought to be specific for PPARgamma may have regulatory effects on inflammatory parameters that are PPARgamma-independent. Future studies of the role of the PPARs in inflammatory and immune responses should include further studies of T cells, T-cell subsets, and dendritic cells but will have to re-examine the issue of PPAR specificity of the ligands being used. This may require further knockout studies and technology, together with the identification of endogenous and perhaps more specific synthetic PPAR ligands. PMID- 11867677 TI - Emerging evidence that molecules expressed by mammalian tissue grafts are recognized by the innate immune system. AB - The innate immune system existed prior to the emergence of adaptive immunity in sharks and higher vertebrates. Homologues of many mammalian innate immune-system elements such as the toll-like receptors exist in species as distant as Drosophila. Selective pressure has led to the development of highly conserved, soluble, and cell-surface receptors that recognize functionally essential molecules shared by microbial pathogens. It is thought that molecular patterns that exquisitely distinguish pathogenic cells from mammalian cells are recognized. Therefore, it would seem unlikely that innate immune-system elements should recognize mammalian tissues. However, there is increasing evidence to suggest that this is the case and that innate immunity promotes rejection of transplanted mammalian tissues, particularly those from other species (xenografts). Evidence for innate recognition of mammalian grafts, the nature of this recognition, and the bi-directional interactions between innate and adaptive immunity that contribute to graft rejection are discussed in this review, with the emphasis on nonvascular xenografts. PMID- 11867679 TI - Enhanced proinflammatory response to endotoxin after priming of macrophages with lead ions. AB - Exposure to lead ions strongly enhances the susceptibility of rodents to endotoxin shock and parasitical infections. Macrophages play a key role during the immune response to lipopolysaccharide (LPS) and during the defense against parasites and might be a target of lead. In the present study, bone marrow derived macrophages (BMMphi) pretreated with lead chloride prior to stimulation with LPS were analyzed for their release of immune mediators. Lead-pretreated cells released up to tenfold increased amounts of tumor necrosis factor-alpha (TNF-alpha), interleukin (IL)-6, IL-12, and prostaglandin E(2) (PGE(2)) but less IL-10 compared with controls. These effects were paralleled by enhanced mRNA levels and were dependent on the duration of lead pretreatment. Inhibition of protein kinase C or of protein synthesis during the priming phase blocked the lead-induced increase of TNF-alpha and IL-6 release. In conclusion, lead ions prime BMMphi for enhanced proinflammatory cytokine secretion in response to LPS, likely by activation of protein kinase C and subsequent synthesis of an unidentified mediator. PMID- 11867678 TI - Thioglycollate peritonitis in mice lacking C5, 5-lipoxygenase, or p47(phox): complement, leukotrienes, and reactive oxidants in acute inflammation. AB - Leukotriene B(4) (LTB(4)) is an easily diffusible proinflammatory chemotactic factor that has been posited to prime the initial inflammatory response for the action of other mediators, including C5a. 5-Lipoxygenase-deficient (5LX(-/-)) and C5-deficient mice only generated about 50% as much peritoneal leukocytosis as wild-type mice following intraperitoneal (IP) challenge with the sterile irritant, thioglycollate (P<0.005). Pretreatment of C5- mice with the specific 5 lipoxygenase inhibitor, zileuton, reduced peritoneal leukocytosis to almost unstimulated levels, suggesting that LTB(4) can act independently of C5a. Previously, LTB(4) and C5a have been shown in vitro to be inactivated by metabolites of superoxide. In the current study, we examined the fate of LTB(4) in the p47(phox-/-) mouse model of chronic granulomatous disease (CGD) in which the phagocyte NADPH oxidase is unable to produce superoxide. p47(phox-/-) mice generated more thioglycollate-elicited peritoneal leukocytosis than wild-type mice. Pretreatment with zileuton caused a 76% reduction in peritoneal leukocytosis in p47(phox-/-) mice (P<0.005) and a 54% reduction in wild-type mice (P<0.05), whereas pretreatment with dexamethasone or toradol (a cyclooxygenase inhibitor) had no effect. Following IP LTB(4) (1 microg/mouse), total recovered peritoneal LTB(4) was similar between p47(phox-/-) and wild-type mice at 10 and 30 min, but was approximately fivefold greater in p47(phox-/-) mice at 180 min. These data suggest that LTB(4) and C5a have separate but overlapping roles in thioglycollate-elicited peritonitis, and at least the leukotriene component is, in turn, regulated by reactive oxidants. PMID- 11867680 TI - Functional characterization of podia formation in normal and malignant hematopoietic cells. AB - Hematopoietic cells extend multiple podia of yet unknown function. Our morphological studies using scanning electron microscopy and functional studies using time-lapse video microscopy suggest that podia formed by CD34+ hematopoietic stem cells (HSC) on the bone marrow stroma component fibronectin are characteristic of lamellipodia at the leading edge and uropodia at the trailing edge, cytoskeletal structures that have previously been shown to be responsible for cell locomotion of lymphocytes. In the leukemic cells studied here, stroma-derived factor-1alpha (SDF-1alpha) led to a significant eightfold increase in transmigration (BCR-ABL-positive BV173 leukemia cell line; P<0.05) and podia formation in all BCR-ABL-positive leukemic cell lines studied (BV173, K562, 32Dp210) and in two of three BCR-ABL-negative lines (HL60, 32D, not KG1a). We could show that SDF-1alpha exposure led to a down-regulation of the gene expression of the chemokine receptors CCR4, CXCR4, and CXCR5, which are associated with cell motility and podia formation, indicating a negative feedback control. In BCR-ABL-positive leukemic cells, the effects of SDF-1alpha on podia formation and cell migration were independent of BCR-ABL-tyrosine kinase activity. Our data are compatible with the hypothesis that formation of specific podia by hematopoietic cells is associated with egression of these cells from the bone marrow. PMID- 11867681 TI - RAGE-mediated neutrophil dysfunction is evoked by advanced glycation end products (AGEs). AB - The accumulation of advanced glycation end products (AGEs) in the tissue and serum of subjects with diabetes has been linked to the pathogenesis of vascular complications. Because diabetes may be also complicated by increased susceptibility to recurrent infection, we investigated the effects of AGEs on human neutrophils, because their burst of activity immediately upon engagement of pathogens or other inflammatory triggers is critical to host response. We demonstrate the presence of receptor for advanced glycation end products (RAGE) at the message and protein levels. We also demonstrate that AGE albumin (but not control albumin) binds with high affinity to human neutrophils (K(d) of 3.7 +/- 0.4 nM). The binding was blocked almost completely by excess soluble RAGE, anti RAGE antibodies, or antibodies to CML-modified albumin. AGE albumin induced a dose-dependent increase in intracellular-free calcium as well as actin polymerization. Further, AGE albumin inhibited transendothelial migration and Staphylococcus aureus-induced but not fMLP-induced production of reactive oxygen metabolite. Moreover, although AGE albumin enhanced neutrophil phagocytosis of S. aureus, it inhibited bacterial killing. We conclude that functional RAGE is present on the plasma membrane of human neutrophils and is linked to Ca(2)(+) and actin polymerization, and engagement of RAGE impairs neutrophil functions. PMID- 11867682 TI - Constitutive and induced expression of DC-SIGN on dendritic cell and macrophage subpopulations in situ and in vitro. AB - DC-SIGN is a C-type lectin, highly expressed on the surface of immature dendritic cells (DCs), that mediates efficient infection of T cells in trans by its ability to bind HIV-1, HIV-2, and SIV. In addition, the ability of DC-SIGN to bind adhesion molecules on surfaces of naive T cells and endothelium also suggests its involvement in T-cell activation and DC trafficking. To gain further insights into the range of expression and potential functions of DC-SIGN, we performed a detailed analysis of DC-SIGN expression in adult and fetal tissues and also analyzed its regulated expression on cultured DCs and macrophages. First, we show that DC-SIGN expression is restricted to subsets of immature DCs in tissues and on specialized macrophages in the placenta and lung. There were no overt differences between DC-SIGN expression in adult and fetal tissues except that DC SIGN expression in alveolar macrophages was only present after birth. Similarly, in tissues, DC-SIGN was observed primarily on immature (CD83-negative) DCs. Secondly, in the peripheral blood, we found expression of DC-SIGN on a small subset of BDCA-2+ plasmacytoid DC precursors (pDC2), concordant with our finding of large numbers of DC-SIGN-positive cells in allergic nasal polyps (previously shown to be infiltrated by DC2). Triple-label confocal microscopy indicated that DC-SIGN was colocalized with BDCA-2 and CD123 on DCs in nasal polyp tissue. Consistent with this finding is our observation that DC-SIGN can be up-regulated on monocyte-derived macrophages upon exposure to the Th2 cytokine, IL-13. In summary, our data demonstrate the relevant populations of DC and macrophages that express DC-SIGN in vivo where it may impact the efficiency of virus infection and indicate that DC-SIGN expression may be involved in the Th2 axis of immunity. PMID- 11867683 TI - Distinct fates of monocytes and T cells directly activated by Pseudomonas aeruginosa exoenzyme S. AB - Gram-negative infections can cause overwhelming inflammatory responses. Although factors other than LPS are clearly involved, these factors and their mechanisms of action have been poorly defined. During studies of LPS-independent inflammatory responses of the gram-negative pathogen Pseudomonas aeruginosa, an important virulence factor (exoenzyme S) was shown to be a potent mitogen for T cells. The current work demonstrates that exoenzyme S selectively induced transcription and secretion of biologically active cytokines and chemokines (chemotactic for neutrophils and T cells) from monocytes. Exoenzyme S stimulated highly purified monocytes independent of T cells. In addition, exoenzyme S stimulated T cells directly; neither T-cell activation (CD69) nor apoptosis (hypodiploidy) required the presence of monocytes. However, T-cell activation was enhanced via a noncontact-dependent mechanism as a result of the secretion of TNF alpha and IL-6. This study identifies a unique property of a gram-negative derived microbial product capable of activating multiple cell types and suggests a mechanism by which exoenzyme S contributes to the immunopathogenesis of cystic fibrosis and sepsis in patients infected with P. aeruginosa. PMID- 11867684 TI - IL-12 suppresses the expression of ocular immunoinflammatory lesions by effects on angiogenesis. AB - Topical application of plasmid DNA encoding IL-12 to the cornea of mice prior to ocular infection with Herpes simplex virus type 1 (HSV) results in diminished corneal immunoinflammatory lesions. Such herpetic stromal keratitis (HSK) reactions in humans represent an important cause of blindness. The effect of IL 12 pretreatment acted via inhibitory effects on corneal neovascularization rather than by inhibiting viral replication or the function of CD4(+) T cells that mediate HSK. The antiangiogenesis induced by IL-12 DNA application was mediated indirectly via the cytokine IFN-gamma and one or both of two chemokine molecules, IP-10 and MIG. Thus IL-12 DNA administration lacked modulatory effects on HSK in GKO mice, indicating the necessary involvement of IFN-gamma induction for antiangiogenesis. In contrast, exposure of GKO mice to IP-10 DNA did suppress the severity of HSK. Furthermore, treatment with specific antisera to IP-10 and MIG in HSV-infected mice abrogated the IL-12-induced inhibitory effect on lesion severity. Taken together, our data indicate that the HSV-induced ocular immunoinflammatory lesions can be modulated by IL-12 and that this effect results from chemokine inhibition of angiogenesis. The use of antiangiogenesis therapy might represent a useful control measure against HSK. PMID- 11867685 TI - Hypertonicity suppresses ionophore-induced product formation and translocation of 5-lipoxygenase in human leukocytes. AB - 5-Lipoxygenase (5-LO) initiates the biosynthesis of proinflammatory leukotrienes from arachidonic acid (AA). Here, we demonstrate that hypertonicity suppresses ionophore-induced 5-LO product formation reversibly in isolated human polymorphonuclear leukocytes (PMNL) and in Mono Mac 6 cells. Hypertonicity blocked the liberation of AA and abrogated translocation of 5-LO to the nuclear membrane. Accordingly, in the presence of exogenous AA, 5-LO product formation was less affected. The effects of hypertonicity were a result of cell shrinkage and not cytosolic hyperosmolarity. Hypertonicity did not inhibit the rapid increase in intracellular Ca(2)(+) induced by ionophores but prevented the ionophore-induced activation of p38 MAPK-regulated MAPKAP kinases, which can phosphorylate and activate 5-LO (and cPLA(2)). In summary, we show that hypertonicity blocks agonist-induced release of AA, 5-LO product formation, and translocation and in parallel, prevents activation of p38 MAPK and downstream 5 LO kinases in leukocytes. PMID- 11867686 TI - Beryllium fluoride-induced cell proliferation: a process requiring P21(ras) dependent activated signal transduction and NF-kappaB-dependent gene regulation. AB - We studied the effect of beryllium fluoride on murine peritoneal macrophages and determined its effects on signal transduction and genetic regulation. At low concentration (1-5 nM), BeF(2) caused an approximate twofold increase in [(3)H]thymidine uptake and cell number, but above 5 nM, it showed cytotoxic effects. BeF(2) increased cellular inositol (1,4,5)trisphosphate (IP(3)) and [Ca(2)(+)](i) about twofold. The rise in [Ca(2)(+)](i) occurred consequent to release from IP(3)-sensitive Ca(2)(+) stores and from influx, mainly via L-type channels. A significant increase in the levels of MEK1, ERK1, p38 MAPK, and JNK phosphorylation was observed in BeF(2)-exposed macrophages. The levels of NF kappaB and CREB transcription factors and the proto-oncogenes c-fos and c-myc were also elevated significantly. Intracellular Ca(2)(+) chelation blocked the effect of BeF(2). We conclude that BeF(2) at low concentration exerts its mitogenic effects in peritoneal macrophages by elevating [Ca(2)(+)](i), which triggers the activation of p21(ras)-dependent MAPK signaling cascades. PMID- 11867687 TI - Engagement of alpha4beta1 integrin by fibronectin induces in vitro resistance of B chronic lymphocytic leukemia cells to fludarabine. AB - B-cell chronic lymphocytic leukemia is characterized by the accumulation of malignant B lymphocytes as a result of abnormal survival signals operating in vivo. Previously, we showed that adhesion of B-CLL cells to the fibronectin fragment H89, a ligand for alpha4beta1 integrin, prevents their spontaneous apoptosis in vitro. We have now studied whether alpha4beta1/H89 interaction affected the response of B-CLL cells to the therapeutic drug fludarabine. B-CLL cells cultured on H89 during treatment with fludarabine showed significantly higher mean viability (P<0.05) than cells cultured on the control polylysine for all doses of drug tested. Similar results were obtained with the EHEB cell line. Analysis of the expression of Bcl-2-family proteins after 48 h of fludarabine treatment revealed that Bcl-xL levels were significantly higher (P<0.05) for cells cultured on H89 than on polylysine and correlated (r=0.56, P<0.05) with the increased cell viability observed on H89 cultures. These results indicate that Bcl-xL is involved in the survival signals induced by alpha4beta1 ligation and may contribute to the progressive drug resistance observed in B-CLL. PMID- 11867688 TI - Toll-like receptor 2 (TLR2) mediates activation of stress-activated MAP kinase p38. AB - Early events in the response of cells to lipopolysaccharide (LPS) include activation of NF-kappaB and stress-activated MAP kinase p38. Recent studies have shown that the human Toll-like receptor 2 (TLR2) mediates activation of NF-kappaB in response to commercial preparations of LPS (comLPS), membrane lipoproteins, and Gram-positive bacterial products. Here, we show that expression of TLR2 in human embryonic kidney 293 cells enabled p38 phosphorylation in response to comLPS, a synthetic bacterial lipoprotein, and B. subtilis. Activation of p38 was confirmed by an in vitro kinase assay using ATF2 as substrate and by an assay measuring activation of the downstream effector of p38, MAP kinase-activated protein kinase in cells. Thus, TLR2 initiated the signaling pathway for p38 in response to bacterial products. PMID- 11867689 TI - Granulocyte-macrophage colony-stimulating factor (GM-CSF)-induced STAT5 activation and target-gene expression during human monocyte/macrophage differentiation. AB - GM-CSF signals through JAK2 and STAT5 and stimulates the expression of STAT5 target genes, such as pim-1 and CIS. Analyzed by EMSA, GM-CSF stimulation led to much stronger STAT5 DNA-binding to pim-1 or CIS GAS elements in primary human monocytes compared with mature macrophages. Similarly, GM-CSF-induced expression of pim-1 and CIS mRNAs was much stronger in monocytes. These differencies were not a result of downregulation of the GM-CSF receptor system or STAT5 expression, because monocytes and macrophages readily expressed GM-CSF receptor, JAK2, STAT5A, and STAT5B mRNAs and proteins. Monocytes expressed significant amounts of truncated STAT5 forms that took part in STAT5-DNA complex formation in GM-CSF stimulated monocytes. This resulted in faster moving STAT5 complexes compared with macrophages in EMSA. Our results demonstrate that STAT5 isoform expression, GM-CSF-induced STAT5 activation, and STAT5 target-gene expression are altered significantly during monocyte/macrophage differentiation. PMID- 11867690 TI - LFA-1 integrin and the microtubular cytoskeleton are involved in the Ca(2)(+) mediated regulation of the activity of the tyrosine kinase PYK2 in T cells. AB - Lymphocyte function-associated antigen (LFA-1) is a member of the beta2 family of integrins that is selectively expressed on leukocytes. Herein, we show that Ca(2)(+) mobilizing agents A23187, thapsigargin, and ionomycin induce an increase in adhesion to the LFA-1 ligand intercellular adhesion molecule-1 (ICAM-1) and activation and redistribution of the proline-rich tyrosine kinase-2 (PYK2) to the microtubule-organizing center (MTOC) in T-lymphoblasts. These effects are similar to those observed upon direct induction of activation of LFA-1 with the stimulatory mAb KIM-127. Most importantly, Ca(2)(+) mobilization did not induce activation of PYK2 when the LFA-1/ICAM-1 interaction was prevented with function blocking mAb, implying that the Ca(2)(+)-induced activation of PYK2 requires integrin engagement. Furthermore, pretreatment of the cells with the Ca(2)(+) chelator EGTA, which depletes the intracellular Ca(2)(+), inhibited the effects of mAb KIM-127 on cell morphology and PYK2 activation. This inhibition with EGTA was not reversed by cross-linking integrin LFA-1 with specific antibodies, indicating that Ca(2)(+) exerts its effects through a target downstream of this integrin. In this regard, immunofluorescence and Western blot analysis showed that Ca(2)(+) chelators affect the organization of the microtubular cytoskeleton and the localization of PYK2 to the MTOC area, suggesting that these agents could inhibit the activation of PYK2 by interfering with the microtubular network of T cells. Taken together, our results demonstrate for the first time an important role for the integrin LFA-1 and the microtubular cytoskeleton in the Ca(2)(+) mediated activation of PYK2 in T-lymphocytes. PMID- 11867691 TI - Beta(3)-mediated engulfment of apoptotic tumor cells by dendritic cells is dependent on CAMKII: inhibition by HIV-1 Tat. AB - In this paper, we show that the engulfment of apoptotic tumor cells by DC requires the activation of the calcium-calmodulin kinase II (CAMKII). Indeed, DC phagocytosis of apoptotic lymphoma cells is consistently inhibited by KN62 and KN93, two blockers of CAMKII, but not by the inactive compound KN92. Wortmannin and LY294002, two inhibitors of the phosphatidyl-inositol-3 kinase, slightly decrease the phagocytosis of apoptotic cells, at variance with PD98059, an inhibitor of the mitogen-activated protein kinase. It is interesting that the addition of synthetic HIV-1 Tat, which we demonstrated to inhibit phagocytosis and calcium influx in DC, blocks the activation of CAMKII elicited via beta(3) integrin, which is involved in apoptotic body engulfment by DC. Experiments performed with Tat-derived peptides showed that this inhibition is mediated by the C-terminal domain of Tat. Finally, pertussis toxin can prevent HIV-1 Tat mediated inhibition, suggesting the involvement of a guanosine triphosphate binding (G) protein in DC-mediated phagocytosis. PMID- 11867692 TI - Toll-like receptor 9 mediates CpG-DNA signaling. AB - Among the bacterial products known to activate the innate immune '1system is bacterial DNA. This activity resides within the nonmethylated CpG motifs of the DNA and is recapitulated using appropriate synthetic CpG containing oligodeoxynucleotides (CpG-ODN). TLR9-deficient mice were shown to exhibit a nonresponsive phenotype-to-bacterial DNA and CpG-ODN. Here, we describe a model system to further characterize CpG-ODN and TLR9 interactions using ectopically expressed TLR9 in HEK293 cells. Expression of TLR9 confers cellular responsiveness to CpG-ODN but not to the other bacterial products. Previous studies identified species-specific CpG-containing sequences; here, we show that expression of murine TLR9 favors responses to CpG-ODN motifs specific to mouse cells, and expression of human TLR9 favors CpG-ODN known to preferentially activate human cells. Response patterns to various CpG-ODN motifs were parallel when cells containing an ectopically expressed TLR9 and endogenous receptor were compared. Here, we also show that TLR9 acts at the cell surface and engages an intracellular signaling pathway that includes MyD88, IRAK, and TRAF6. PMID- 11867693 TI - Activation of Arabidopsis vacuolar processing enzyme by self-catalytic removal of an auto-inhibitory domain of the C-terminal propeptide. AB - Vacuolar processing enzyme (VPE) is a cysteine proteinase responsible for the maturation of various vacuolar proteins in higher plants. To clarify the mechanism of maturation and activation of VPE, we expressed the precursors of Arabidopsis gamma VPE in insect cells. The cells accumulated a glycosylated proprotein precursor (pVPE) and an unglycosylated preproprotein precursor (ppVPE) which might be unfolded. The N-terminal sequence of pVPE revealed that ppVPE had a 22-amino-acid signal peptide to be removed co-translationally. Under acidic conditions, the 56-kDa pVPE was self-catalytically converted to a 43-kDa intermediate form (iVPE) and then to the 40-kDa mature form (mVPE). N-terminal sequencing of iVPE and mVPE showed that sequential removal of the C-terminal propeptide and N-terminal propeptide produced mVPE. Both iVPE and mVPE exhibited the activity, while pVPE exhibited no activity. These results imply that the removal of the C-terminal propeptide is essential for activating the enzyme. Further removal of the N-terminal propeptide from iVPE is not required to activate the enzyme. To demonstrate that the C-terminal propeptide functions as an inhibitor of VPE, we expressed the C-terminal propeptide and produced specific antibodies against it. We found that the C-terminal propeptide reduced the activity of VPE and that this inhibitory activity was suppressed by specific antibodies against it. Our findings suggest that the C-terminal propeptide functions as an auto-inhibitory domain that masks the catalytic site. Thus, the removal of the C-terminal propeptide of pVPE might expose the catalytic site of the enzyme. PMID- 11867694 TI - Cloning of gibberellin 3 beta-hydroxylase cDNA and analysis of endogenous gibberellins in the developing seeds in watermelon. AB - We have isolated Cv3h, a cDNA clone from the developing seeds of watermelon, and have demonstrated significant amino acid homology with gibberellin (GA) 3 beta hydroxylases. This cDNA clone was expressed in Escherichia coli as a fusion protein that oxidized GA(9) and GA(12) to GA(4) and GA(14), respectively. The Cv3h protein had the highest similarity with pumpkin GA 2 beta,3 beta hydroxylase, but did not possess 2 beta-hydroxylation function. RNA blot analysis showed that the gene was expressed primarily in the inner parts of developing seeds, up to 10 d after pollination (DAP). In the parthenocarpic fruits induced by treatment with 1-(2-chloro-4-pyridyl)-3-phenylurea (CPPU), the embryo and endosperm of the seeds were undeveloped, whereas the integumental tissues, of maternal origin, showed nearly normal development. Cv3h mRNA was undetectable in the seeds of CPPU-treated fruits, indicating that the GA 3 beta-hydroxylase gene was expressed in zygotic cells. In our analysis of endogenous GAs from developing seeds, GA(9) and GA(4) were detected at high levels but those of GA(20) and GA(1) were very low. This demonstrates that GA biosynthesis in seeds prefers a non-13 hydroxylation pathway over an early 13-hydroxylation pathway. We also analyzed endogenous GAs from seeds of the parthenocarpic fruits. The level of bioactive GA(4 )was much lower there than in normal seeds, indicating that bioactive GAs, unconnected with Cv3h, exist in integumental tissues during early seed development. PMID- 11867695 TI - Molecular characterization of a phosphoenolpyruvate carboxylase from a thermophilic cyanobacterium, Synechococcus vulcanus with unusual allosteric properties. AB - A gene for phosphoenolpyruvate carboxylase (PEPC) was isolated from a thermophilic cyanobacterium, Synechococcus vulcanus, by screening a genomic DNA library using the coding region of Anacystis nidulans 6301 PEPC as a probe. The S. vulcanus PEPC gene (SvPEPC) had an open reading frame for a polypeptide of 1,011 amino acid residues with a calculated molecular mass of 116.4 kDa. SvPEPC was expressed in E. coli BL21 Codonplus (DE3), using pET32a as a vector. The purified recombinant SvPEPC protein with a tag showed a single band of 120 kDa on SDS-PAGE. The enzyme forms homotetramer as judged by gel filtration. SvPEPC retained full activity even after incubation at 50 degrees C for 60 min or exposure to 0.5 M guanidine-HCl at 30 degrees C for 20 h, being more stable than C4-form PEPC from Zea mays (ZmPEPC(C4)). SvPEPC activity showed a sharp optimum temperature of 42 degrees C at pH 7.5 and an optimum pH of 9.0 at 30 degrees C. The enzyme, unlike most plant PEPCs, was predominantly activated by fructose 1,6 bisphosphate (Fruc-1,6-P(2)), and slightly stimulated by 3-phosphoglycerate (3 PGA), glucose 6-phosphate (Gluc-6-P), glucose 1-phosphate, Glu and Gln. Acetyl CoA known as a strong activator of most bacterial PEPCs but not of plant PEPCs, showed no effect on the enzyme activity. SvPEPC was more sensitive to the inhibition by Asp at higher pH (9.0) than lower pH (7.0), contrary to Coccochloris peniocystis PEPC and plant PEPCs. I(0.5) for Asp was increased about 2-fold by Gluc-6-P while markedly decreased by Fruc-1,6-P(2), Glu and Gln about 3 to 4-fold. The regulation mechanism of SvPEPC is not readily interpretable by conventional allosteric models. PMID- 11867696 TI - Recovery of photosynthetic systems during rewetting is quite rapid in a terrestrial cyanobacterium, Nostoc commune. AB - Recovery processes of photosynthetic systems during rewetting were studied in detail in a terrestrial, highly drought-tolerant cyanobacterium, Nostoc commune. With absorption of water, the weight of N. commune colony increased in three phases with half-increase times of about 1 min, 2 h and 9 h. Fluorescence intensities of phycobiliproteins and photosystem (PS) I complexes were recovered almost completely within 1 min, suggesting that their functional forms were restored very quickly. Energy transfer from allophycocyanin to the core-membrane linker peptide (L(CM)) was recovered within 1 min, but not that from L(CM) to PSII. PSI activity and cyclic electron flow around PSI recovered within 2 min, while the PSII activity recovered in two phases after a time lag of about 5 min, with half times of about 20 min and 2 h. Photosynthetic CO(2) fixation was restored almost in parallel with the first recovery phase of the PSII reaction center activity. Although the amount of absorbed water became more than 20 times the initial dry weight of the N. commune colony in the presence of sufficient water, about twice the initial dry weight was enough for recovery and maintenance of the PSII activity. PMID- 11867697 TI - A gene encoding multidrug resistance (MDR)-like protein is induced by aluminum and inhibitors of calcium flux in wheat. AB - A cDNA clone exclusively induced by aluminum (Al) was isolated from root apices of wheat (Triticum aestivum L.) by the differential display method. The predicted amino acid sequence exhibited homology to the multidrug resistance (MDR) proteins that is known as a member of the ATP-binding cassette (ABC) protein superfamily. Thus this gene was named TaMDR1 (Triticum aestivum MDR). TaMDR1 was induced as a function of Al concentration in the range from 5 to 50 microM, which is in the range of Al content in natural acid soil environment. The concentration required for the induction was lower in the Al-sensitive cultivar than in the Al-tolerant cultivar, indicating that the accumulation of TaMDR1 mRNA was associated with the events caused by Al toxicity rather than Al tolerance. TaMDR1 was significantly induced by the exposure to lanthanum, gadolinium and ruthenium red, which are known as inhibitors of calcium channels. Furthermore, decreasing of calcium ion in growth medium caused stimulation of the gene expression. These results suggested that the induction of TaMDR1 is caused by the breaking of calcium homeostasis which occurred at early stage of Al toxicity. PMID- 11867698 TI - Toward understanding the different function of two types of parenchyma cells in bamboo culms. AB - The bamboo, woody monocot, has two types of parenchyma cells in the ground tissues of its culm, in contrast to a single type of parenchyma cell in rice, maize and other major crop species. The distribution of cell wall components, including lignin, (1-->3), (1-->4)-beta-D-glucans (MGs), the highly-substituted glucuronoarabinoxylans (hsGAXs) and low-branched xylans (lbXs) in ground parenchyma tissue of Phyllostachys heterocycla var. pubescens culms was studied at various developmental stages using light microscopy (LM), UV-microscopy, transmission electron microscopy (TEM) and immunolabeling techniques. The short parenchyma cell walls were lignified in 2-month-old bamboo culms just as the long parenchyma cell walls were. The lignified regions were confined to the portions in contact with the long parenchyma cell walls, while the walls at the cell corner region never lignified, even in 7-year-old culms. Significant differences were also found in the hemicellulose distribution between the short and long parenchyma cell walls. In bamboo parenchyma tissue, MGs were localized in short parenchyma cell walls and few were found in long parenchyma cell walls in both young and 7-year-old culms. The distribution of hsGAXs was similar to that of MGs in young culms, but they only appeared in the cell corner region of short parenchyma cells in old culms. Low-branched xylans were distributed in the lignified, but not in unlignified parenchyma cell walls. Based on this evidence, the differences of function in both short and long parenchyma cells in a bamboo culm are discussed. PMID- 11867699 TI - Enhanced susceptibility of photosynthesis to low-temperature photoinhibition due to interruption of chill-induced increase of S-adenosylmethionine decarboxylase activity in leaves of spinach (Spinacia oleracea L.). AB - The possible involvement of polyamines in the chilling tolerance of spinach (Spinacia oleracea L.) was investigated focusing on photosynthesis. During chilling at 8/5C (day/night) for 6 d, S-adenosylmethionine decarboxylase (SAMDC) activity increased significantly in leaves in parallel with the increase in putrescine and spermidine (Spd) content in leaves and chloroplasts. Treatment of leaves with methylglyoxal-bis(guanylhydrazone) (MGBG), an SAMDC inhibitor, resulted in the deterioration of plant growth and photosynthesis under chilling conditions, which was reversed by the concomitant treatment with Spd through the roots. Plants treated with MGBG showed lower photochemical efficiency of PSII than either the control or plants treated with MGBG plus Spd during chilling and even after transfer to warm conditions, suggesting an increase of photoinhibition due to low Spd in chloroplasts. Indeed, MGBG-treated plants had much lower activities of thylakoid electron transport and enzymes in carbon metabolism as well as higher degrees of lipid peroxidation of thylakoid membranes compared to the control. These results indicate that the enhanced activity of SAMDC with a consequential rise of Spd in chloroplasts is crucial for the cold acclimation of the photosynthetic apparatus in spinach leaves. PMID- 11867701 TI - A high temperature-sensitive mutant of Synechococcus sp. PCC 7002 with modifications in the endogenous plasmid, pAQ1. AB - To study thermal adaptations in the cyanobacterium, Synechococcus sp. PCC 7002, we screened about 3,000 mutants for their tolerance to high temperature, and found one, SHT1, that is sensitive to high-temperature stress. The mutant had a modified gene construct in the endogenous plasmid, pAQ1. One of the four ORFs, ORF93, was duplicated, and its mRNA level was higher than in the wild type. At 38 degrees C, the growth of SHT1 was retarded as compared with the wild type, and above 38 degrees C, almost all the cells of SHT1 died. This temperature is much lower than that required for induction of heat shock proteins. Interestingly, in both the wild type and SHT1, the thermal stability of oxygen-evolving machinery increased upon acclimation to high temperatures. These findings indicate that the lack of thermal tolerance in the SHT1 strain is likely independent of the adaptation of the PSII complex and heat shock responses, whereas there are essential contributions of genes in the endogenous plasmid to the adaptation to high temperature. Thus, understanding the role of pAQ1 in the adaptation of Synechococcus sp. PCC 7002 to high-temperature environments is the first step in elucidating the function of this plasmid. PMID- 11867700 TI - Aluminum-induced rapid changes in the microtubular cytoskeleton of tobacco cell lines. AB - Aluminum (Al) is a major factor that limits plant growth in acid soils. It causes a cessation of root growth and changes in root morphology suggesting a role of the root cytoskeleton as a target of Al-toxicity. Here we report a rapid effect of Al on the microtubular cytoskeleton of the suspension tobacco cell lines BY-2 and VBI-0. Viability studies showed that the cells were more sensitive to Al during exponential phase as compared to stationary cells. During the first hours of exposure, Al induced the formation of additional bundles of cortical microtubules (cMTs), whereas the thickness of the individual bundles decreased. Prolonged exposure resulted in disorientation of cMTs. These changes of cMTs preceded the decrease of cell viability by several hours and were accompanied by an increase in the levels of alpha-tubulin (in its tyrosinated form) and elements of the tubulin-folding chaperone CCT. These findings suggest that the microtubular cytoskeleton is one of the early targets of Al toxicity. PMID- 11867702 TI - Gamma-tubulin in barley and tobacco: sequence relationship and RNA expression patterns in developing leaves during mitosis and post-mitotic growth. AB - gamma-Tubulin is typically associated with microtubule organising centres, such as the centrosome, and appears to mediate microtubule nucleation. Centrosomes are usually not found in higher plants, but active genes homologous to gamma-tubulin have been identified in the plant kingdom, including the angiosperms Arabidopsis, maize and rice. We have isolated and characterised gamma-tubulin cDNA sequences of two further angiosperm species, barley and tobacco. Sequence comparison revealed a phylogenetic tree with distinct clusters corresponding to the systematic position of the species. Furthermore, domains, thought to be exposed in the folded protein and to be candidates for interaction with associated, nucleation-site related proteins, exhibited motifs highly specific of multicellular plants. Strong expression of the gamma-tubulin genes, as determined by Northern blotting, correlated with mitotic activity. Expression dropped distinctly when mitotic activity ceased. Thus, in post-mitotic tissues that showed intricate reshuffling of cortical microtubule arrays related to cell shaping only very low gamma-tubulin steady-state RNA levels were found, contrasting with the situation for alpha-tubulin. The findings indicate that gamma-tubulin expression in plants may be more tightly linked to mitosis, although there is some gamma-tubulin expression at the RNA level even after mitosis. It follows that the post-mitotic changes in microtubular arrays may be less dependent on concurrent gamma-tubulin RNA expression than mitotic cells. PMID- 11867703 TI - Characterization of tobacco MADS-box genes involved in floral initiation. AB - MADS-box genes encode regulatory factors that are involved at various stages in plant development. These genes function not only during early floral meristem identity, but also when the fate of floral organ primordia is determined in a later step. Here, we screened a floral bud cDNA library to isolate a tobacco MADS box gene, NtMADS4, using the rice MADS-box gene, OsMADS1, as a probe. We previously reported that OsMADS1 plays a critical role in flower development in rice. Ectopic expression of NtMADS4 caused phenotypes of extremely early flowering as well as dwarfism. Plant MADS proteins have a K domain that mediates the formation of dimers. This dimerization appears to be an essential step for a functional protein complex. NtMADS11 was isolated as an interacting partner of NtMADS4 by yeast two-hybrid screening. The latter was included in the AGAMOUS like 2 (AGL2) family whereas the former was categorized in the SQUAMOSA (SQUA) family. While the transcript of NtMADS4 was detectable only in reproductive organs, that of NtMADS11 was seen in both reproductive and vegetative organs. Expression levels were high for both genes during early developmental stages. Ectopic expression of NtMADS11 and OsMADS14 was able to rescue the floral organ defects seen in the strong ap1-1 mutant. Roles of NtMADS4 and NtMADS11 in the floral initiation are discussed. PMID- 11867704 TI - Brassinosteroids control the proliferation of leaf cells of Arabidopsis thaliana. AB - The growth of leaves in the model plant, Arabidopsis thaliana (L.) Heynh., is determined by the extent of expansion of individual cells and by cell proliferation. Mutants of A. thaliana with known defects in the biosynthesis or perception of brassinosteroids develop small leaves. When the leaves of brassinosteroid-related mutants, det2 (de-etiolated2 = cro1) and dwf1 (dwarf1 = cro2) were compared to wild-type plants, an earlier cessation of leaf expansion was observed; a detailed anatomical analysis further revealed that the mutants had fewer cells per leaf blade. Treatment of the det2 mutants with the brassinosteroid, brassinolide, reversed the mutation and restored the potential for growth to that of the wild type. Restoration of leaf size could not be explained solely on the basis of an increase in individual cell volume, thus suggesting that brassinosteroids play a dual role in regulating cell expansion and proliferation. PMID- 11867705 TI - High-quality life extension by the enzyme peptide methionine sulfoxide reductase. AB - Cumulative oxidative damages to cell constituents are considered to contribute to aging and age-related diseases. The enzyme peptide methionine sulfoxide reductase A (MSRA) catalyzes the repair of oxidized methionine in proteins by reducing methionine sulfoxide back to methionine. However, whether MSRA plays a role in the aging process is poorly understood. Here we report that overexpression of the msrA gene predominantly in the nervous system markedly extends the lifespan of the fruit fly Drosophila. The MSRA transgenic animals are more resistant to paraquat-induced oxidative stress, and the onset of senescence-induced decline in the general activity level and reproductive capacity is delayed markedly. The results suggest that oxidative damage is an important determinant of lifespan, and MSRA may be important in increasing the lifespan in other organisms including humans. PMID- 11867706 TI - Revised estimates of human cochlear tuning from otoacoustic and behavioral measurements. AB - We develop an objective, noninvasive method for determining the frequency selectivity of cochlear tuning at low and moderate sound levels. Applicable in humans at frequencies of 1 kHz and above, the method is based on the measurement of stimulus-frequency otoacoustic emissions and, unlike previous noninvasive physiological methods, does not depend on the frequency selectivity of masking or suppression. The otoacoustic measurements indicate that at low sound levels human cochlear tuning is more than twice as sharp as implied by standard behavioral studies and has a different dependence on frequency. New behavioral measurements designed to minimize the influence of nonlinear effects such as suppression agree with the emission-based values. A comparison of cochlear tuning in cat, guinea pig, and human indicates that, contrary to common belief, tuning in the human cochlea is considerably sharper than that found in the other mammals. The sharper tuning may facilitate human speech communication. PMID- 11867707 TI - Conserved regulation of the lymphocyte-specific expression of lck in the Fugu and mammals. AB - The lck gene encodes a lymphocyte-specific protein-tyrosine kinase that is implicated in T cell maturation and signaling. In mammals, the transcription of the lck gene is regulated by two independent promoters, the proximal promoter, which is active in thymocytes, and the distal promoter, which dominates in mature T cells. In the human and mouse lck gene loci, the two promoter elements are separated by at least 40 kb and 10 kb, respectively. In this study, we have cloned and sequenced 60 kb from the pufferfish (Fugu rubripes) lck locus. The promoter region of the Fugu lck spans only 4.2 kb and contains a proximal and a distal promoter in the 2.3-kb region adjacent to the coding sequence. By generating transgenic mice, we have demonstrated that the compact promoter of the Fugu lck contains regulatory elements that direct expression to lymphoid organs of mice. We were able to localize the regulatory elements to a short region of 830 bp without losing specificity to cultured human T cell line. These results show that the basic mechanisms that mediate lymphocyte-specific expression are conserved between teleosts and mammals. The short promoter of the Fugu lck isolated by us offers a powerful tool for labeling T cells, targeting expression, and manipulating T cell activity in fishes as well as in mammals. PMID- 11867708 TI - Structural basis for the guanine nucleotide-binding activity of tissue transglutaminase and its regulation of transamidation activity. AB - Tissue transglutaminase (TG) is a Ca2+-dependent acyltransferase with roles in cellular differentiation, apoptosis, and other biological functions. In addition to being a transamidase, TG undergoes a GTP-binding/GTPase cycle even though it lacks any obvious sequence similarity with canonical GTP-binding (G) proteins. Guanine nucleotide binding and Ca2+ concentration reciprocally regulate TG's transamidation activity, with nucleotide binding being the negative regulator. Here we report the x-ray structure determined to 2.8-A resolution of human TG complexed with GDP. Although the transamidation active site is similar to those of other known transglutaminases, the guanine nucleotide-binding site of TG differs markedly from other G proteins. The structure suggests a structural basis for the negative regulation of transamidation activity by bound nucleotide, and the positive regulation of transamidation by Ca2+. PMID- 11867709 TI - Framing effects and risky decisions in starlings. AB - Animals are predominantly risk prone toward reward delays and risk averse toward reward amounts. Humans in turn tend to be risk-seeking for losses and risk averse for gains. To explain the human results, Prospect Theory postulates a convex utility for losses and concave utility for gains. In contrast, Scalar Utility Theory (SUT) explains the animal data by postulating that the cognitive representation of outcomes follows Weber's Law, namely that the spread of the distribution of expected outcomes is proportional to its mean. SUT also would explain human results if utility (even if it is linear on expected outcome) followed Weber's Law. We present an experiment that simulates losses and gains in a bird, the European Starling, to test the implication of SUT that risk proneness/aversion should extend to any aversive/desirable dimension other than time and amount of reward. Losses and gains were simulated by offering choices of fixed vs. variable outcomes with lower or higher outcomes than what the birds expected. The subjects were significantly more risk prone for losses than for gains but, against expectations, they were not significantly risk averse toward gains. The results are thus, in part, consistent with Prospect Theory and SUT and show that risk attitude in humans and birds may obey a common fundamental principle. PMID- 11867710 TI - Alpha-helical stabilization by side chain shielding of backbone hydrogen bonds. AB - We study atomic models of the thermodynamics of the structural transition of peptides that form alpha-helices. The effect of sequence variation on alpha-helix formation for alanine-rich peptides, Ac-Ala21-methyl amide (A21) and Ac-A5 (AAARA)3A-methyl amide (Fs peptide), is investigated by atomic simulation studies of the thermodynamics of the helix-coil transition in explicit water. The simulations show that the guanidinium group in the Arg side chains in the Fs peptide interacts with the carbonyl group four amino acids upstream in the chain and desolvates backbone hydrogen bonds. This desolvation can be directly correlated with a higher probability of hydrogen bond formation. We find that Fs has higher helical content than A21 at all temperatures. A small modification in the amber force field reproduces the experimental helical content and helix-coil transition temperatures for the Fs peptide. PMID- 11867712 TI - Nitric oxide down-regulates brain-derived neurotrophic factor secretion in cultured hippocampal neurons. AB - The regulation of neurotrophin (NT) secretion is critical for many aspects of NT mediated neuronal plasticity. Neurons release NTs by activity-regulated secretion pathways, initiated either by neurotransmitters and/or by existing NTs by a positive-feedback mechanism. This process depends on calcium release from intracellular stores. Little is known, however, about potential pathways that down-regulate NT secretion. Here we demonstrate that nitric oxide (NO) induces a rapid down-regulation of brain-derived neurotrophic factor (BDNF) secretion in cultured hippocampal neurons. Similar effects occur by activating a downstream target of intracellular NO, the soluble guanylyl cyclase, or by increasing the levels of its product, cGMP. Furthermore, down-regulation of BDNF secretion is mediated by cGMP-activated protein kinase G, which prevents calcium release from inositol 1,4,5-trisphosphate-sensitive stores. Our data indicate that the NO/cGMP/protein kinase G pathway represents a signaling mechanism by which neurons can rapidly down-regulate BDNF secretion and suggest that, in hippocampal neurons, NT secretion is finely tuned by both stimulatory and inhibitory signals. PMID- 11867711 TI - Neurotoxin-induced degeneration of dopamine neurons in Caenorhabditis elegans. AB - Parkinson's disease is a complex neurodegenerative disorder characterized by the death of brain dopamine neurons. In mammals, dopamine neuronal degeneration can be triggered through exposure to neurotoxins accumulated by the presynaptic dopamine transporter (DAT), including 6-hydroxydopamine (6-OHDA) and 1-methyl-4 phenylpyridinium. We have established a system for the pharmacological and genetic evaluation of neurotoxin-induced dopamine neuronal death in Caenorhabditis elegans. Brief (1 h) exposure of green fluorescent protein-tagged, living worms to 6-OHDA causes selective degeneration of dopamine neurons. We demonstrate that agents that interfere with DAT function protect against 6-OHDA toxicity. 6-OHDA-triggered neural degeneration does not require the CED-3/CED-4 cell death pathway, but is abolished by the genetic disruption of the C. elegans DAT. PMID- 11867714 TI - Dynamics of bacterial phenotype selection in a colonized host. AB - The population dynamics of Helicobacter pylori during colonization in an infected animal host provide a quantifiable experimental model of in vivo microbial phenotype evolution. Phenotype variability in H. pylori populations can be typed as polymorphic expression of Lewis antigens on their cell surfaces. The high mutational frequency of H. pylori for Lewis expression provides substrate for differential selection by the host. Experimental challenge and successful colonization of mice and gerbils allows tracking of H. pylori phenotype variability from the initial inoculation to the ultimate establishment of a quasispecies. Colonization data provide a quantitative experimental model of phenotype evolution in a relatively large population (>10(4) individuals) over a relatively long evolutionary time scale (>10(3) generations). A mathematical model is developed to interpret the data in terms of the dynamic processes occurring during colonization. The mathematical model distinguishes the roles of selection and mutation; quantifies the effects of initial phenotype diversity, mutational frequency, and selective advantage; and applies generally to phenotype evolution in biological populations. PMID- 11867713 TI - Cytotoxic T lymphocyte antigen-4 (CTLA-4) limits the expansion of encephalitogenic T cells in experimental autoimmune encephalomyelitis (EAE) resistant BALB/c mice. AB - We and others previously reported that cytotoxic T lymphocyte antigen-4 (CTLA-4) regulates the severity of peptide-induced experimental autoimmune encephalomyelitis (EAE) in mouse strains that are inherently susceptible to the disease. In this report, we show that CTLA-4 engagement also controls disease susceptibility in BALB/c mice, a strain considered to be resistant to EAE induction. Although immunization of BALB/c mice with syngeneic spinal cord homogenate or an I-A(d)-binding myelin peptide antigen failed to result in EAE, immunization with either antigen preparation in conjunction with anti-CTLA-4 resulted in both clinical and histological EAE. CTLA-4 blockade also resulted in a preferential increase in the frequency of antigen-specific T cells secreting IFN-gamma. We conclude that CTLA-4 controls susceptibility in BALB/c mice by limiting the expansion of autoreactive T cells present in the periphery, suggesting a mechanism whereby CTLA-4 contributes to the maintenance of peripheral T cell tolerance to self antigens. PMID- 11867715 TI - Whole-gene APC deletions cause classical familial adenomatous polyposis, but not attenuated polyposis or "multiple" colorectal adenomas. AB - Familial adenomatous polyposis (FAP) is a dominantly inherited colorectal tumor predisposition that results from germ-line mutations in the APC gene (chromosome 5q21). FAP shows substantial phenotypic variability: classical polyposis patients develop more than 100 colorectal adenomas, whereas those with attenuated polyposis (AAPC) have fewer than 100 adenomas. A further group of individuals, so called "multiple" adenoma patients, have a phenotype like AAPC, with 3-99 polyps throughout the colorectum, but mostly have no demonstrable germ-line APC mutation. Routine mutation detection techniques fail to detect a pathogenic APC germ-line mutation in approximately 30% of patients with classical polyposis and 90% of those with AAPC/multiple adenomas. We have developed a real-time quantitative multiplex PCR assay to detect APC exon 14 deletions. When this technique was applied to a set of 60 classical polyposis and 143 AAPC/multiple adenoma patients with no apparent APC germ-line mutation, deletions were found exclusively in individuals with classical polyposis (7 of 60, 12%). Fine-mapping of the region suggested that the majority (6 of 7) of these deletions encompassed the entire APC locus, confirming that haploinsufficiency can result in a classical polyposis phenotype. Screening for germ-line deletions in APC mutation negative individuals with classical polyposis seems warranted. PMID- 11867716 TI - Models of the extracellular domain of the nicotinic receptors and of agonist- and Ca2+-binding sites. AB - We constructed a three-dimensional model of the amino-terminal extracellular domain of three major types of nicotinic acetylcholine receptor, (alpha7)5, (alpha4)2(beta2)3, and (alpha1)2beta1gammadelta, on the basis of the recent x-ray structure determination of the molluscan acetylcholine-binding protein. Comparative analysis of the three models reveals that the agonist-binding pocket is much more conserved than the overall structure. Differences exist, however, in the side chains of several residues. In particular, a phenylalanine residue, present in beta2 but not in alpha7, is proposed to contribute to the high affinity for agonists in receptors containing the beta2 subunit. The semiautomatic docking of agonists in the ligand-binding pocket of (alpha7)5 led to positions consistent with labeling and mutagenesis experiments. Accordingly, the quaternary ammonium head group of nicotine makes a pi-cation interaction with W148 (alpha7 numbering), whereas the pyridine ring is close to both the cysteine pair 189-190 and the complementary component of the binding site. The intrinsic affinities inferred from docking give a rank order epibatidine > nicotine > acetylcholine, in agreement with experimental values. Finally, our models offer a structural basis for potentiation by external Ca2+. PMID- 11867717 TI - Experimentally based model of a complex between a snake toxin and the alpha 7 nicotinic receptor. AB - To understand how snake neurotoxins interact with nicotinic acetylcholine receptors, we have elaborated an experimentally based model of the alpha cobratoxin-alpha7 receptor complex. This model was achieved by using (i) a three dimensional model of the alpha7 extracellular domain derived from the crystallographic structure of the homologous acetylcholine-binding protein, (ii) the previously solved x-ray structure of the toxin, and (iii) nine pairs of residues identified by cycle-mutant experiments to make contacts between the alpha-cobratoxin and alpha7 receptor. Because the receptor loop F occludes entrance of the toxin binding pocket, we submitted this loop to a dynamics simulation and selected a conformation that allowed the toxin to reach its binding site. The three-dimensional structure of the toxin-receptor complex model was validated a posteriori by an additional double-mutant experiment. The model shows that the toxin interacts perpendicularly to the receptor axis, in an equatorial position of the extracellular domain. The tip of the toxin central loop plugs into the receptor between two subunits, just below the functional receptor loop C, the C-terminal tail of the toxin making adjacent additional interactions at the receptor surface. The receptor establishes major contacts with the toxin by its loop C, which is assisted by principal (loops A and B) and complementary (loops D, F, and 1) functional regions. This model explains the antagonistic properties of the toxin toward the neuronal receptor and opens the way to the design of new antagonists. PMID- 11867718 TI - Impaired retention of spatial memory after transection of longitudinally oriented axons of hippocampal CA3 pyramidal cells. AB - Longitudinally oriented axon collaterals of CA3 pyramidal cells may be critical for integrating distributed information in the hippocampus. To investigate the possible role of this pathway in the retention of spatial memory, we made a single transversely oriented cut through the dorsal CA3 region of each hippocampus. Although the lesion involved <3% of the hippocampal volume, it nonetheless disrupted memory retention in a water maze in preoperatively trained rats. New learning in a different water maze was attenuated. No significant impairment occurred in rats with longitudinally oriented cuts, or in animals with ibotenic acid-induced lesions of similar magnitude. To characterize the effect of a focal lesion on the integrity of longitudinally projecting axons, we stained degenerating cells and fibers in rats with unilateral CA3 transections by using FluoroJade-B. Degenerating terminals were seen across a wide region posterior to the cut, and were present in the strata of areas CA3 and CA1 that are innervated by CA3 pyramidal cells. These results suggest that the integrity of longitudinally oriented, translamellar axons of CA3 pyramidal cells may be necessary for efficient acquisition and retention of spatial memory. PMID- 11867719 TI - Visualization of functionally activated circuitry in the brain. AB - We have used a transgenic approach to visualize functionally activated neurons and their projections. The transgenic mice contain a tau-lacZ fusion gene regulated by the promoter for c-fos, an immediate early gene that is rapidly induced in neurons after functional stimulation. Constitutive expression of beta galactosidase (beta-gal), the lacZ product, was low and in accord with previous reports of c-fos expression. However, expression of beta-gal in positive neurons was clearly in cell bodies, axons, and dendrites. Treatment of the mice with kainic acid, a strong inducer of c-fos expression, resulted in high induction of beta-gal. beta-gal was induced in the same defined populations of neurons in the brain as those that express c-fos after kainic acid induction. Furthermore, the pattern of beta-gal expression within the neurons changed over time after kainic acid treatment. Early after kainate treatment, beta-gal was found mainly in cell bodies; at later times, expression extended further along the neuronal processes. This expression pattern is consistent with induction and anterograde transport of the Fos-Tau-beta-gal protein in the neurons. To test whether a functionally activated pathway could be visualized, transgenic mice were deprived of water, which activates nuclei involved in body fluid homeostasis. beta-gal induction was traced in neurons and their processes in the lamina terminalis, in magnocellular neurons of the supraoptic and paraventricular nuclei, and in their projections to the posterior pituitary gland. This strategy allowed the mapping of an activated osmoregulatory pathway. This transgenic approach may have general application in the mapping of functionally activated circuitry in the brain. PMID- 11867720 TI - Regulation of transferrin-mediated iron uptake by HFE, the protein defective in hereditary hemochromatosis. AB - The protein defective in hereditary hemochromatosis, called HFE, is similar to MHC class I-type proteins and associates with beta2-microglobulin (beta2M). Its association with beta2M was previously shown to be necessary for its stability, normal intracellular processing, and cell surface expression in transfected COS cells. Here we use stably transfected Chinese hamster ovary cell lines expressing both HFE and beta2M or HFE alone to study the effects of beta2M on the stability and maturation of the HFE protein and on the role of HFE in transferrin receptor 1 (TfR1)-mediated iron uptake. In agreement with prior studies on other cell lines, we found that overexpression of HFE, without overexpressing beta2M, resulted in a decrease in TfR1dependent iron uptake and in lower iron levels in the cells, as evidenced by ferritin and TfR1 levels measured at steady state. However, overexpression of both HFE and beta2M had the reverse effect and resulted in an increase in TfR1-dependent iron uptake and increased iron levels in the cells. The HFE-beta2M complex did not affect the affinity of TfR1 for transferrin or the internalization rate of transferrin-bound TfR1. Instead, HFE beta2M enhanced the rate of recycling of TfR1 and resulted in an increase in the steady-state level of TfR1 at the cell surface of stably transfected cells. We propose that Chinese hamster ovary cells provide a model to explain the effect of the HFE-beta2M complex in duodenal crypt cells, where the HFE-beta2M complex appears to facilitate the uptake of transferrin-bound iron to sense the level of body iron stores. Impairment of this process in duodenal crypt cells leads them to be iron poor and to signal the differentiating enterocytes to take up iron excessively after they mature into villus cells in the duodenum of hereditary hemochromatosis patients. PMID- 11867722 TI - Network of coupled promoting motions in enzyme catalysis. AB - A network of coupled promoting motions in the enzyme dihydrofolate reductase is identified and characterized. The present identification is based on genomic analysis for sequence conservation, kinetic measurements of multiple mutations, and mixed quantum/classical molecular dynamics simulations of hydride transfer. The motions in this network span time scales of femtoseconds to milliseconds and are found on the exterior of the enzyme as well as in the active site. This type of network has broad implications for an expanded role of the protein fold in catalysis as well as ancillaries such as the engineering of altered protein function and the action of drugs distal to the active site. PMID- 11867721 TI - Genomic DNA breakpoints in AML1/RUNX1 and ETO cluster with topoisomerase II DNA cleavage and DNase I hypersensitive sites in t(8;21) leukemia. AB - The translocation t(8;21)(q22;q22) is one of the most frequent chromosome translocations in acute myeloid leukemia (AML). AML1/RUNX1 at 21q22 is involved in t(8;21), t(3;21), and t(16;21) in de novo and therapy-related AML and myelodysplastic syndrome as well as in t(12;21) in childhood B cell acute lymphoblastic leukemia. Although DNA breakpoints in AML1 and ETO (at 8q22) cluster in a few introns, the mechanisms of DNA recombination resulting in t(8;21) are unknown. The correlation of specific chromatin structural elements, i.e., topoisomerase II (topo II) DNA cleavage sites, DNase I hypersensitive sites, and scaffold-associated regions, which have been implicated in chromosome recombination with genomic DNA breakpoints in AML1 and ETO in t(8;21) is unknown. The breakpoints in AML1 and ETO were clustered in the Kasumi 1 cell line and in 31 leukemia patients with t(8;21); all except one had de novo AML. Sequencing of the breakpoint junctions revealed no common DNA motif; however, deletions, duplications, microhomologies, and nontemplate DNA were found. Ten in vivo topo II DNA cleavage sites were mapped in AML1, including three in intron 5 and seven in intron 7a, and two were in intron 1b of ETO. All strong topo II sites colocalized with DNase I hypersensitive sites and thus represent open chromatin regions. These sites correlated with genomic DNA breakpoints in both AML1 and ETO, thus implicating them in the de novo 8;21 translocation. PMID- 11867723 TI - Effect of the human cytomegalovirus IE86 protein on expression of E2F-responsive genes: a DNA microarray analysis. AB - We have previously reported that the immediate early (IE)-86 protein of human cytomegalovirus (HCMV) pushes the cell cycle toward S phase but inhibits cell division [Murphy, E. A., Streblow, D. N., Nelson, J. A. & Stinski, M. F. (2000) J. Virol. 74, 7108-7118]. We determined the cellular genes activated by the IE86 protein in permissive human fibroblast cells. A 4-fold or greater increase in the steady-state RNA from many cellular genes that regulate the cell cycle, the enzymes for DNA precursor synthesis, and the initiation of cellular DNA replication was detected by high-density DNA microarray analysis. Northern blot analysis confirmed the DNA microarray data. The viral IE86 protein induced a significant increase in the cellular steady-state RNA level from the B-myb, cyclin E, cdk-2, E2F-1, ribonucleotide reductase 1, ribonucleotide reductase 2, thymidylate synthetase, MCM3, and MCM7 genes, but actin RNA was not affected. Cellular genes regulated by the E2F transcription factors were strongly activated by the IE86 protein. In most cases, the cellular genes induced by the IE86 protein were also induced by HCMV infection. This study demonstrates the global array of cellular genes activated by the IE86 protein that pushes progression of the cell cycle from G0/G1 toward the G1/S transition point. PMID- 11867724 TI - Rapid topology mapping of Escherichia coli inner-membrane proteins by prediction and PhoA/GFP fusion analysis. AB - We present an approach that allows rapid determination of the topology of Escherichia coli inner-membrane proteins by a combination of topology prediction and limited fusion-protein analysis. We derive new topology models for 12 inner membrane proteins: MarC, PstA, TatC, YaeL, YcbM, YddQ, YdgE, YedZ, YgjV, YiaB, YigG, and YnfA. We estimate that our approach should make it possible to arrive at highly reliable topology models for roughly 10% of the approximately 800 inner membrane proteins thought to exist in E. coli. PMID- 11867725 TI - Cortex mapping reveals regionally specific patterns of genetic and disease specific gray-matter deficits in twins discordant for schizophrenia. AB - The symptoms of schizophrenia imply disruption to brain systems supporting higher order cognitive activity, but whether these systems are impacted differentially against a background of diffuse cortical gray-matter deficit remains ambiguous. Some unaffected first-degree relatives of schizophrenics also manifest cortical gray-matter deficits, but it is unclear whether these changes are isomorphic with those in patients, and the answer is critical to understanding the neurobiological conditions necessary for disease expression given a predisposing genotype. Here we report three-dimensional cortical surface maps (probabilistic atlases matching subjects' anatomy point by point throughout cortex) in monozygotic (MZ) and dizygotic (DZ) twins discordant for chronic schizophrenia along with demographically matched control twins. A map encoding the average differences between schizophrenia patients and their unaffected MZ co-twins revealed deficits primarily in dorsolateral prefrontal cortex, superior temporal gyrus, and superior parietal lobule. A map encoding variation associated with genetic proximity to a patient (MZ co-twins > DZ co-twins > control twins) isolated deficits primarily in polar and dorsolateral prefrontal cortex. In each case, the statistical significance was confirmed through analysis of 10,000 Monte Carlo permutations, and the remaining cortex was shown to be significantly less affected by contrast analysis. The disease-related deficits in gray matter were correlated with measures of symptom severity and cognitive dysfunction but not with duration of illness or antipsychotic drug treatment. Genetic and disease specific influences thus affect gray matter in partially nonoverlapping areas of predominantly heteromodal association cortex, changes that may act synergistically in producing overt behavioral features of the disorder. PMID- 11867726 TI - Interaction with glycosaminoglycans is required for cyclophilin B to trigger integrin-mediated adhesion of peripheral blood T lymphocytes to extracellular matrix. AB - Cyclophilins A and B (CyPA and CyPB) are cyclosporin A-binding proteins that are involved in inflammatory events. We have reported that CyPB interacts with two types of cell-surface-binding sites. The first site corresponds to a functional receptor and requires interaction with the central core of CyPB. This region is highly conserved in cyclophilins, suggesting that CyPA and CyPB might share biological activities mediated by interaction with this receptor. The second site is identified with glycosaminoglycans (GAGs), the binding region located in the N terminus of CyPB. The difference in the N-terminal extensions of CyPA and CyPB suggests that a unique interaction with GAGs might account for selective activity of CyPB. To explore this hypothesis, we analyzed the lymphocyte responses triggered by CyPA, CyPB, and CyPB(KKK-), a mutant unable to interact with GAGs. The three ligands seemed capable enough to elicit calcium signal and chemotaxis by binding to the same signaling receptor. In contrast, only CyPB enhanced firm adhesion of T cells to the extracellular matrix. This activity depended on the interactions with GAGs and signaling receptor. CyPB-mediated adhesion required CD147 presumably because it was a costimulatory molecule and was related to an activation of alpha4beta1 and alpha4beta7 integrins. Finally, we showed that CyPB was capable mainly to enhance T cell adhesion of the CD4+CD45RO+ subset. The present data indicate that CyPB rather than CyPA is a proinflammatory factor for T lymphocytes and highlight the crucial role of CyPB-GAG interaction in the chemokine-like activity of this protein. PMID- 11867727 TI - Survival and regeneration of rubrospinal neurons 1 year after spinal cord injury. AB - Scientific interest to find a treatment for spinal cord injuries has led to the development of numerous experimental strategies to promote axonal regeneration across the spinal cord injury site. Although these strategies have been developed in acute injury paradigms and hold promise for individuals with spinal cord injuries in the future, little is known about their applicability for the vast majority of paralyzed individuals whose injury occurred long ago and who are considered to have a chronic injury. Some studies have shown that the effectiveness of these approaches diminishes dramatically within weeks after injury. Here we investigated the regenerative capacity of rat rubrospinal neurons whose axons were cut in the cervical spinal cord 1 year before. Contrary to earlier reports, we found that rubrospinal neurons do not die after axotomy but, rather, they undergo massive atrophy that can be reversed by applying brain derived neurotrophic factor to the cell bodies in the midbrain. This administration of neurotrophic factor to the cell body resulted in increased expression of growth-associated protein-43 and Talpha1 tubulin, genes thought to be related to axonal regeneration. This treatment promoted the regeneration of these chronically injured rubrospinal axons into peripheral nerve transplants engrafted at the spinal cord injury site. This outcome is a demonstration of the regenerative capacity of spinal cord projection neurons a full year after axotomy. PMID- 11867728 TI - Activity-dependent isolation of the presenilin- gamma -secretase complex reveals nicastrin and a gamma substrate. AB - Presenilin heterodimers apparently contain the active site of gamma-secretase, a polytopic aspartyl protease involved in the transmembrane processing of both the Notch receptor and the amyloid-beta precursor protein. Although critical to embryonic development and the pathogenesis of Alzheimer's disease, this protease is difficult to characterize, primarily because it is a multicomponent complex of integral membrane proteins. Here the functional gamma-secretase complex was isolated by using an immobilized active site-directed inhibitor of the protease. Presenilin heterodimers and nicastrin bound specifically to this inhibitor under conditions tightly correlating with protease activity, whereas several other presenilin-interacting proteins (beta-catenin, calsenilin, and presenilin associated protein) did not bind. Moreover, anti-nicastrin antibodies immunoprecipitated gamma-secretase activity from detergent-solubilized microsomes. Unexpectedly, C83, the major endogenous amyloid-beta precursor protein substrate of gamma-secretase, was also quantitatively associated with the complex. These results provide direct biochemical evidence that nicastrin is a member of the active gamma-secretase complex, indicate that beta-catenin, calsenilin, and presenilin-associated protein are not required for gamma activity, and suggest an unprecedented mechanism of substrate-protease interaction. PMID- 11867730 TI - Heritability of lobar brain volumes in twins supports genetic models of cerebral laterality and handedness. AB - Although the left and right human cerebral hemispheres differ both functionally and anatomically, little is known about the environmental or genetic factors that govern central nervous system asymmetry. Nevertheless, cerebral asymmetry is strongly correlated with handedness, and handedness does have a significant genetic component. To explore the relative contribution of environmental and genetic influences on cerebral asymmetry, we examined the volumes of left and right cerebral cortex in a large cohort of aging identical and fraternal twins and explored their relationship to handedness. Cerebral lobar volumes had a major genetic component, indicating that genes play a large role in changes in brain volume that occur with aging. Shared environment, which likely represents in utero events, had about twice the effect on the left hemisphere as on the right, consistent with less genetic control over the left hemisphere. To test the major genetic models of handedness and cerebral asymmetry, twin pairs were divided into those with two right handers and those with at least one left hander (nonright handers). Genetic factors contributed twice the influence to left and right cerebral hemispheric volumes in right-handed twin pairs, suggesting a large decrement in genetic control of cerebral volumes in the nonright-handed twin pairs. This loss of genetic determination of the left and right cerebral hemispheres in the nonright-handed twin pairs is consistent with models postulating a right-hand/left-hemisphere-biasing genetic influence, a "right shift" genotype that is lost in nonright handers, resulting in decreased cerebral asymmetry. PMID- 11867729 TI - Persistence of oligodendrocyte precursor cells and altered myelination in optic nerve associated to retina degeneration in mice devoid of all thyroid hormone receptors. AB - Thyroid hormone (3,5,3'-triiodo-l-thyronine or T3) exerts a pleiotropic activity during central nervous system development. Hypothyroidism during the fetal and postnatal life results in an irreversible mental retardation syndrome. At the cellular level, T3 is known to act on neuronal and glial lineages and to control cell proliferation, apoptosis, migration, and differentiation. Oligodendrocyte precursor cells (OPC) found at birth in the optic nerves are self-renewing cells that normally differentiate during the first 3 weeks of rodent postnatal life into postmitotic myelinating oligodendrocytes. In vitro, the addition of T3 to OPC is sufficient to trigger their terminal differentiation. The present analysis of T3 receptor knockout mice reveals that the absence of all T3 receptor results in the persistence of OPC proliferation in adult optic nerves, in a default in myelination, and sometimes in the degeneration of the retinal ganglion neurons. Thus, T3 signaling is necessary in vivo to promote the complete differentiation of OPC. PMID- 11867731 TI - Cloning of nitroalkane oxidase from Fusarium oxysporum identifies a new member of the acyl-CoA dehydrogenase superfamily. AB - The flavoprotein nitroalkane oxidase (NAO) from Fusarium oxysporum catalyzes the oxidation of nitroalkanes to the respective aldehydes with production of nitrite and hydrogen peroxide. The sequences of several peptides from the fungal enzyme were used to design oligonucleotides for the isolation of a portion of the NAO gene from an F. oxysporum genomic DNA preparation. This sequence was used to clone the cDNA for NAO from an F. oxysporum cDNA library. The sequence of the cloned cDNA showed that NOA is a member of the acyl-CoA dehydrogenase (ACAD) superfamily. The members of this family share with NAO a mechanism that is initiated by proton removal from carbon, suggesting a common chemical reaction for this superfamily. NAO was expressed in Escherichia coli and the recombinant enzyme was characterized. Recombinant NAO has identical kinetic parameters to enzyme isolated from F. oxysporum but is isolated with oxidized FAD rather than the nitrobutyl-FAD found in the fungal enzyme. NAO purified from E. coli or from F. oxysporum has no detectable ACAD activity on short- or medium-chain acyl CoAs, and medium-chain acyl-CoA dehydrogenase and short-chain acyl-CoA dehydrogenase are unable to catalyze oxidation of nitroalkanes. PMID- 11867732 TI - Members of the PIAS family act as SUMO ligases for c-Jun and p53 and repress p53 activity. AB - The activity of the p53 tumor suppressor protein and the c-Jun protooncogene is regulated by posttranslational modifications, such as phosphorylation or ubiquitination. In addition, covalent attachment of the ubiquitin-like modifier SUMO appears to modulate their transcriptional activity. Sumoylation proceeds via an enzymatic pathway that is mechanistically analogous to ubiquitination, but requires a different E1-activating enzyme and Ubc9, a SUMO-specific E2 conjugating enzyme. Here, we show that two members of the PIAS family, PIAS1 and PIASxbeta, act as specific E3-like ligases that promote sumoylation of p53 and c Jun in vitro and in vivo. The PIAS proteins physically interact with both p53 and c-Jun. In addition, they bind to Ubc9, suggesting that they recruit the E2 enzyme to their respective substrate. The SUMO ligase activity requires the conserved zinc-finger domain, which is distantly related to the essential RING-finger motif, found in a subset of ubiquitin ligases. Furthermore, similar to RING-type ubiquitin ligases, PIASxbeta can catalyze its own modification. Hence, these data further extend the analogy between the ubiquitin and SUMO pathway. Strikingly, PIAS proteins strongly repress the transcriptional activity of p53, suggesting that the PIAS-SUMO pathway plays a crucial role in the regulation of p53 and presumably other transcription factors. PMID- 11867733 TI - Quantitative in vivo assessment of lung microstructure at the alveolar level with hyperpolarized 3He diffusion MRI. AB - The study of lung emphysema dates back to the beginning of the 17th century. Nevertheless, a number of important questions remain unanswered because a quantitative localized characterization of emphysema requires knowledge of lung structure at the alveolar level in the intact living lung. This information is not available from traditional imaging modalities and pulmonary function tests. Herein, we report the first in vivo measurements of lung geometrical parameters at the alveolar level obtained with 3He diffusion MRI in healthy human subjects and patients with severe emphysema. We also provide the first experimental data demonstrating that 3He gas diffusivity in the acinus of human lung is highly anisotropic. A theory of anisotropic diffusion is presented. Our results clearly demonstrate substantial differences between healthy and emphysematous lung at the acinar level and may provide new insights into emphysema progression. The technique offers promise as a clinical tool for early diagnosis of emphysema. PMID- 11867735 TI - The mammalian oocyte orchestrates the rate of ovarian follicular development. AB - The development of both the mammalian oocyte and the somatic cell compartments of the ovarian follicle is highly coordinated; this coordination ensures that the ovulated oocyte is ready to undergo fertilization and subsequent embryogenesis. Disruption of this synchrony results in oocyte developmental failure. Communication between the oocyte and companion somatic cells is essential for successful development of both follicular compartments. However, it was not previously known whether one cell type, either the somatic or the germ cell compartment, determines the overall rate of follicular development. To test the hypothesis that the oocyte orchestrates the rate of follicle development, mid sized oocytes isolated from secondary follicles were transferred back to primordial follicles, the earliest stage of follicular development. This transfer doubled the rate of follicular development and the differentiation of follicular somatic cells. Oocyte development in these accelerated follicles appeared normal; recovered oocytes were competent to undergo fertilization and embryonic development. These results demonstrate that oocytes orchestrate and coordinate the development of mammalian ovarian follicles and that the rate of follicular development is based on a developmental program intrinsic to the oocyte. PMID- 11867734 TI - Thyroid hormone is a critical determinant for the regulation of the cochlear motor protein prestin. AB - The most impressive property of outer hair cells (OHCs) is their ability to change their length at high acoustic frequencies, thus providing the exquisite sensitivity and frequency-resolving capacity of the mammalian hearing organ. Prestin, a protein related to a sulfate/anion transport protein, recently has been identified and proposed as the OHC motor molecule. Homology searches of 1.5 kb of genomic DNA 5' of the coding region of the prestin gene allowed the identification of a thyroid hormone (TH) response element (TRE) in the first intron upstream of the prestin ATG codon. Prestin(TRE) bound TH receptors as a monomer or presumptive heterodimer and mediated a triiodothyronine-dependent transactivation of a heterologous promotor in response to triiodothyronine receptors alpha and beta. Retinoid X receptor-alpha had an additive effect. Expression of prestin mRNA and prestin protein was reduced strongly in the absence of TH. Although prestin protein typically was redistributed to the lateral membrane before the onset of hearing, an immature pattern of prestin protein distribution across the entire OHC membrane was noted in hypothyroid rats. The data suggest TH as a first transcriptional regulator of the motor protein prestin and as a direct or indirect modulator of subcellular prestin distribution. PMID- 11867736 TI - Enhanced signaling through the IL-2 receptor in CD8+ T cells regulated by antigen recognition results in preferential proliferation and expansion of responding CD8+ T cells rather than promotion of cell death. AB - Multiple cytokines, including IL-2, can affect T cell proliferation and survival. However, IL-2 can lead to apoptosis as well as proliferation, making unclear whether IL-2 receptor (IL-2R) signals ultimately have a predominantly positive or negative effect. To address this issue, we examined the effect of enhancing IL-2R signals in CD8(+) T cells after antigen stimulation by engineering a transgenic (Tg) mouse strain with CD8(+) T cells capable of augmented, regulated, autocrine IL-2R signaling after target recognition by means of expression of a chimeric granulocyte-macrophage colony-stimulating factor (GM-CSF)/IL-2R. The Tg CD8(+) T cells can bind the granulocyte-macrophage colony-stimulating GM-CSF produced by antigen stimulation, but the GM-CSF binding results in delivery of an IL-2R signal. After antigen stimulation in vivo, the Tg T cells demonstrated marked increases in the initial proliferative response and cell expansion and displayed continued increases in cell expansion after repeated antigen exposure. These data suggest that the predominant role of IL-2R signals delivered to responding CD8(+) T cells is to set the size of the initial response to antigen by promoting T cell proliferation and survival and not cell death. PMID- 11867737 TI - Functional recovery following traumatic spinal cord injury mediated by a unique polymer scaffold seeded with neural stem cells. AB - To better direct repair following spinal cord injury (SCI), we designed an implant modeled after the intact spinal cord consisting of a multicomponent polymer scaffold seeded with neural stem cells. Implantation of the scaffold neural stem cells unit into an adult rat hemisection model of SCI promoted long term improvement in function (persistent for 1 year in some animals) relative to a lesion-control group. At 70 days postinjury, animals implanted with scaffold plus-cells exhibited coordinated, weight-bearing hindlimb stepping. Histology and immunocytochemical analysis suggested that this recovery might be attributable partly to a reduction in tissue loss from secondary injury processes as well as in diminished glial scarring. Tract tracing demonstrated corticospinal tract fibers passing through the injury epicenter to the caudal cord, a phenomenon not present in untreated groups. Together with evidence of enhanced local GAP-43 expression not seen in controls, these findings suggest a possible regeneration component. These results may suggest a new approach to SCI and, more broadly, may serve as a prototype for multidisciplinary strategies against complex neurological problems. PMID- 11867738 TI - Analysis of the ultraviolet B response in primary human keratinocytes using oligonucleotide microarrays. AB - UV radiation is the most important environmental skin aggressor, causing cancer and other problems. This paper reports the use of oligonucleotide microarray technology to determine changes in gene expression in human keratinocytes after UVB treatment. Examination of the effects of different doses at different times after irradiation gave a global picture of the keratinocyte response to this type of insult. Five hundred thirty-nine regulated transcripts were found and organized into nine different clusters depending on behavior patterns. Classification of these genes into 23 functional categories revealed that several biological processes are globally affected by UVB. In addition to confirming a majority up-regulation of the transcripts related to the UV-specific inflammatory and stress responses, significant increases were seen in the expression of genes involved in basal transcription, splicing, and translation as well as in the proteasome-mediated degradation category. On the other hand, those transcripts belonging to the metabolism and adhesion categories were strongly downregulated. These results demonstrate the complexity of the transcriptional profile of the UVB response, describe several cellular processes previously not known to be affected by UV irradiation, and serve as a basis for the global characterization of UV-regulated genes and pathways. PMID- 11867739 TI - Genes, pseudogenes, and Alu sequence organization across human chromosomes 21 and 22. AB - Human chromosomes 21 and 22 (mainly the q-arms) were the first complete parts of the human genome released. Our analysis of genes, pseudogenes (Psig), and Alu repeats across these chromosomes include the following findings: The number of gene structures containing untranslated exons exceeds 25%; the terminal exon tends to be the largest among exons, whereas, the initial intron tends to be the largest among introns; single-exon gene length is approximately the mean gene exon number times the mean internal exon length; processed Psig lengths are on average approximately the same as single-exon gene length; and the G+C content and length of genes are uncorrelated. The counts and distribution of genes, Psig, and Alu sequences and G+C variation are evaluated with respect to clusters and overdispersions. Other assessments concern comparisons of intergenic lengths, properties of Psig sequences, and correlations between Alu and Psig sequences. PMID- 11867740 TI - Isolation of a diterpenoid substance with potent neuroprotective activity from fetal calf serum. AB - Excess activation of glutamate receptors and production of free radicals including nitric oxide may result in severe and irreversible damage to the mammalian central nervous system (CNS), but endogenous defense systems that protect neurons from these insults are poorly understood. Here, we purified and isolated a neuroprotective substance, which has been named "serofendic acid," from a lipophilic fraction of FCS based on the ability to protect rat primary cortical neurons against nitric oxide cytotoxicity. Mass spectrometry and NMR spectroscopy revealed the chemical structure of serofendic acid (15-hydroxy-17 methylsulfinylatisan-19-oic acid) as a sulfur-containing atisane-type diterpenoid, which is unique among known endogenous substances. Synthetic serofendic acid exhibited potent protective actions on cortical neurons against cytotoxicity of a nitric oxide donor as well as of glutamate, although it did not show appreciable influences on glutamate receptor-mediated responses in these neurons. Electron spin resonance analysis demonstrated that serofendic acid had no direct scavenging activity on nitric oxide radicals but was capable of inhibiting the generation of hydroxyl radical, a presumed "executor" radical in the nitric oxide-mediated neurotoxic cascade. These findings suggest that serofendic acid is a low-molecular-weight bioactive factor that promotes survival of CNS neurons, probably through the attenuation of free radical-mediated insults. PMID- 11867741 TI - Helix formation via conformation diffusion search. AB - The helix-coil transition kinetics of an alpha-helical peptide were investigated by time-resolved infrared spectroscopy coupled with laser-induced temperature jump initiation method. Specific isotope labeling of the amide carbonyl groups with 13C at selected residues was used to obtain site-specific information. The relaxation kinetics following a temperature jump, obtained by probing the amide I' band of the peptide backbone, exhibit nonexponential behavior and are sensitive to both initial and final temperatures. These data are consistent with a conformation diffusion process on the folding energy landscape, in accord with a recent molecular dynamics simulation study. PMID- 11867743 TI - Invariant chain induces B cell maturation in a process that is independent of its chaperonic activity. AB - Early stages of B cell development take place in the bone marrow, resulting in formation of immature B cells, which migrate to the spleen for their final differentiation into mature cells. This final maturation step is essential for B cells to become responsive to antigens and to participate in the immune response. Previously, we showed that the MHC class II chaperone, invariant chain (Ii), controls the differentiation of B cells from the immature to the mature stage. In this study, by generating transgenic mice expressing truncated Ii lacking its luminal domain, we could dissect the chaperonin activity of Ii from its role in B cell maturation. We demonstrate in vivo that Ii N-terminal domain is directly involved in the maturation of B cells and is sufficient to promote B cell differentiation. PMID- 11867744 TI - The human pathogen Pseudomonas aeruginosa utilizes conserved virulence pathways to infect the social amoeba Dictyostelium discoideum. AB - Genetically accessible host models are useful for studying microbial pathogenesis because they offer the means to identify novel strategies that pathogens use to evade immune mechanisms, cause cellular injury, and induce disease. We have developed conditions under which the human pathogen Pseudomonas aeruginosa infects Dictyostelium discoideum, a genetically tractable eukaryotic organism. When D. discoideum is plated on nutrient agar plates with different P. aeruginosa strains, the bacteria form lawns on these plates with amoebae embedded in them. Virulent P. aeruginosa strains kill these amoebae and leave an intact bacterial lawn. A number of P. aeruginosa mutants have been identified that are avirulent in this assay. Amoebae feed on these bacteria and form plaques in their bacterial lawns. One avirulent mutant strain carries an insertional mutation in the lasR gene. LasR is a transcription factor that controls a number of virulence genes in a density-dependent fashion. Another class of avirulent P. aeruginosa mutants is defective in type III secretion. One mutant lacks the PscJ protein, a structural component of the secretion apparatus, suggesting that cytotoxins are injected into the D. discoideum cell. One of these cytotoxins is ExoU, and exoU mutants are avirulent toward D. discoideum. Complementation of the lasR and exoU mutations restores virulence. Therefore, P. aeruginosa uses conserved virulence pathways to kill D. discoideum. PMID- 11867742 TI - The antitumor histone deacetylase inhibitor suberoylanilide hydroxamic acid exhibits antiinflammatory properties via suppression of cytokines. AB - Suberoylanilide hydroxamic acid (SAHA) is a hydroxamic acid-containing hybrid polar molecule; SAHA specifically binds to and inhibits the activity of histone deacetylase. Although SAHA, like other inhibitors of histone deacetylase, exhibits antitumor effects by increasing expression of genes regulating tumor survival, we found that SAHA reduces the production of proinflammatory cytokines in vivo and in vitro. A single oral administration of SAHA to mice dose dependently reduced circulating TNF-alpha, IL-1-beta, IL-6, and IFN-gamma induced by lipopolysaccharide (LPS). Administration of SAHA also reduced hepatic cellular injury in mice following i.v. injection of Con A. SAHA inhibited nitric oxide release in mouse macrophages stimulated by the combination of TNF-alpha plus IFN gamma. Human peripheral blood mononuclear cells stimulated with LPS in the presence of SAHA released less TNF-alpha, IL-1-beta, IL-12, and IFN-gamma (50% reduction at 100-200 nM). The production of IFN-gamma stimulated by IL-18 plus IL 12 was also inhibited by SAHA (85% at 200 nM). However, SAHA did not affect LPS induced synthesis of the IL-1-beta precursor, the IL-1 receptor antagonist, or the chemokine IL-8. In addition, IFN-gamma induced by anti-CD3 was not suppressed by SAHA. Steady-state mRNA levels for LPS-induced TNF-alpha and IFN-gamma in peripheral blood mononuclear cells were markedly decreased, whereas IL-8 and IL-1 beta mRNA levels were unaffected. Because SAHA exhibits antiinflammatory properties in vivo and in vitro, inhibitors of histone deacetylase may stimulate the expression of genes that control the synthesis of cytokines and nitric oxide or hyperacetylate other targets. PMID- 11867745 TI - Thyroid hormone activates oligodendrocyte precursors and increases a myelin forming protein and NGF content in the spinal cord during experimental allergic encephalomyelitis. AB - Remyelination in the adult central nervous system has been demonstrated in different experimental models of demyelinating diseases. However, there is no clear evidence that remyelination occurs in multiple sclerosis, the most diffuse demyelinating disease. In this article, we explore the possibility of promoting myelination in experimental allergic encephalomyelitis, a widely used experimental model of multiple sclerosis, by recruiting progenitors and channeling them into oligodendroglial lineage through administration of thyroid hormone (T4). A large number of proliferating cells (BrdUrd uptake and Ki67-IR) and the expression of markers for undifferentiated precursors (nestin) increased in the subventricular zone and spinal cord of experimental allergic encephalomyelitis animals. T4 administration reduces proliferation and nestin immunoreactivity and up-regulates expression of markers for oligodendrocyte progenitors [polysialylated-neural cell adhesion molecule (PSA-NCAM), O4, A2B5] and mature oligodendrocytes (myelin basic protein) in the spinal cord, olfactory bulb, and subventricular zone. PMID- 11867746 TI - NQO1 stabilizes p53 through a distinct pathway. AB - Wild-type p53 is a tumor-suppressor gene that encodes a short-lived protein that, upon accumulation, induces growth arrest or apoptosis. Accumulation of p53 occurs mainly by posttranslational events that inhibit its proteosomal degradation. We have reported previously that inhibition of NAD(P)H: quinone oxidoreductase 1 (NQO1) activity by dicoumarol induces degradation of p53, indicating that NQO1 plays a role in p53 stabilization. We now have found that wild-type NQO1, but not the inactive polymorphic NQO1, can stabilize endogenous as well as transfected wild-type p53. NQO1-mediated p53 stabilization was especially prominent under induction of oxidative stress. NQO1 also partially inhibited p53 degradation mediated by the human papilloma virus E6 protein, but not when mediated by Mdm-2. Inhibitors of heat shock protein 90 (hsp90), radicicol and geldanamycin, induced degradation of p53 and suppressed p53-induced apoptosis in normal thymocytes and myeloid leukemic cells. Differences in the effectiveness of dicoumarol and hsp90 inhibitors to induce p53 degradation and suppress apoptosis in these cell types indicate that NQO1 and hsp90 stabilize p53 through different mechanisms. Our results indicate that NQO1 has a distinct role in the regulation of p53 stability, especially in response to oxidative stress. The present data on the genetic and pharmacologic regulation of the level of p53 have clinical implications for tumor development and therapy. PMID- 11867747 TI - Melanin-concentrating hormone 1 receptor-deficient mice are lean, hyperactive, and hyperphagic and have altered metabolism. AB - Melanin-concentrating hormone (MCH) is a cyclic 19-aa hypothalamic neuropeptide derived from a larger prohormone precursor of MCH (Pmch), which also encodes neuropeptide EI (NEI) and neuropeptide GE (NGE). Pmch-deficient (Pmch-/-) mice are lean, hypophagic, and have an increased metabolic rate. Transgenic mice overexpressing Pmch are hyperphagic and develop mild obesity. Consequently, MCH has been implicated in the regulation of energy homeostasis. The MCH 1 receptor (MCH1R) is one of two recently identified G protein-coupled receptors believed to be responsible for the actions of MCH. We evaluated the physiological role of MCH1R by generating MCH1R-deficient (Mch1r-/-) mice. Mch1r-/- mice have normal body weights, yet are lean and have reduced fat mass. Surprisingly, Mch1r-/- mice are hyperphagic when maintained on regular chow, and their leanness is a consequence of hyperactivity and altered metabolism. Consistent with the hyperactivity, Mch1r-/- mice are less susceptible to diet-induced obesity. Importantly, chronic central infusions of MCH induce hyperphagia and mild obesity in wild-type mice, but not in Mch1r-/- mice. We conclude that MCH1R is a physiologically relevant MCH receptor in mice that plays a role in energy homeostasis through multiple actions on locomotor activity, metabolism, appetite, and neuroendocrine function. PMID- 11867748 TI - GATA-1 binding sites mapped in the beta-globin locus by using mammalian chIp-chip analysis. AB - The expression of the beta-like globin genes is intricately regulated by a series of both general and tissue-restricted transcription factors. The hemapoietic lineage-specific transcription factor GATA-1 is important for erythroid differentiation and has been implicated in regulating the expression of the erythroid-specific genes including the genes of the beta-globin locus. In the human erythroleukemic K562 cell line, only one DNA region has been identified previously as a putative site of GATA-1 interaction by in vivo footprinting studies. We mapped GATA-1 binding throughout the beta-globin locus by using chIp chip analysis of K562 cells. We found that GATA-1 binds in a region encompassing the HS2 core element, as was previously identified, and an additional region of GATA-1 binding upstream of the gammaG gene. This approach will be of general utility for mapping transcription factor binding sites within the beta-globin locus and throughout the genome. PMID- 11867749 TI - The peroxisome proliferator-activated receptor delta, an integrator of transcriptional repression and nuclear receptor signaling. AB - The three PPAR (peroxisome proliferator-activated receptor) isoforms are critical regulators of lipid homeostasis by controlling the balance between the burning and storage of long chain fatty acids. Whereas PPARalpha and PPARgamma have been studied extensively, the function of PPARdelta remains the most elusive. Intriguingly, in cotransfection experiments, PPARdelta is a potent inhibitor of ligand-induced transcriptional activity of PPARalpha and PPARgamma. This inhibition is achieved, in part, by binding of PPARdelta to a peroxisome proliferator response element and the association of nonliganded PPARdelta with corepressors SMRT (silencing mediator for retinoid and thyroid hormone receptors), SHARP (SMRT and histone deacetylase-associated repressor protein), and class I histone deacetylases. Stable expression of PPARdelta inhibits the expression of endogenous PPARalpha target gene expression in 3T3-PPARalpha cells, whereas a PPARdelta mutant that does not interact with the corepressor SMRT loses its ability to repress the induction of PPARalpha target gene. Similarly, stable expression of PPARdelta in 3T3-PPARgamma cells leads to inhibition of PPARgamma target gene expression and PPARgamma-mediated adipogenesis. Given the widespread expression of PPARdelta and the restricted pattern for PPARalpha and PPARgamma, these results suggest a role for PPARdelta as a gateway receptor whose relative levels of expression can be used to modulate PPARalpha and PPARgamma activity. PMID- 11867750 TI - Apoptosis and melanogenesis in human melanoma cells induced by anthrax lethal factor inactivation of mitogen-activated protein kinase kinase. AB - Lethal factor, the principal virulence factor of Bacillus anthracis, inhibits mitogen-activated protein kinase (MAPK) signaling by proteolytically cleaving MAPK kinases. Edema factor, another component of anthrax toxin, is an adenylate cyclase, which increases intracellular cAMP. Inhibition of MAPK signaling with either anthrax lethal toxin (LeTx) or small molecule MAPK kinase inhibitors triggers apoptosis in human melanoma cells. Normal melanocytes do not undergo apoptosis in response to MAPK inhibition but arrest in the G1 phase of the cell cycle. Importantly, in vivo treatment of human melanoma xenograft tumors in athymic nude mice with LeTx results in significant or complete tumor regression without apparent side effects, suggesting that inhibiting the MAPK signaling pathway may be a useful strategy for treating melanoma. Additionally, interrupting MAPK signaling with LeTx and elevating cAMP with anthrax edema toxin in both melanoma cells and melanocytes lead to dramatic melanin production, perhaps explaining the formation of blackened eschars in cutaneous anthrax. PMID- 11867751 TI - Structural views of the ligand-binding cores of a metabotropic glutamate receptor complexed with an antagonist and both glutamate and Gd3+. AB - Crystal structures of the extracellular ligand-binding region of the metabotropic glutamate receptor, complexed with an antagonist, (S)-(alpha)-methyl-4 carboxyphenylglycine, and with both glutamate and Gd3+ ion, have been determined by x-ray crystallographic analyses. The structure of the complex with the antagonist is similar to that of the unliganded resting dimer. The antagonist wedges the protomer to maintain an inactive open form. The glutamate/Gd3+ complex is an exact 2-fold symmetric dimer, where each bi-lobed protomer adopts the closed conformation. The surface of the C-terminal domain contains an acidic patch, whose negative charges are alleviated by the metal cation to stabilize the active dimeric structure. The structural comparison between the active and resting dimers suggests that glutamate binding tends to induce domain closing and a small shift of a helix in the dimer interface. Furthermore, an interprotomer contact including the acidic patch inhibited dimer formation by the two open protomers in the active state. These findings provide a structural basis to describe the link between ligand binding and the dimer interface. PMID- 11867752 TI - Ex vivo cell labeling with 64Cu-pyruvaldehyde-bis(N4-methylthiosemicarbazone) for imaging cell trafficking in mice with positron-emission tomography. AB - We have used copper-64-pyruvaldehyde-bis(N4-methylthiosemicarbazone) (64Cu-PTSM) to radiolabel cells ex vivo for in vivo positron-emission tomography (PET) imaging studies of cell trafficking in mice and for eventual application in patients. 2-[18F]-Fluoro-2-deoxy-d-glucose (FDG) cell labeling also was evaluated for comparison. 64Cu-PTSM uptake by C6 rat glioma (C6) cells increased for 180 min and then stabilized. The labeling efficiency was directly proportional to 64Cu-PTSM concentration and influenced negatively by serum. Label uptake per cell was greater with 64Cu-PTSM than with FDG. However, both 64Cu-PTSM- and FDG labeled cells showed efflux of cell activity into supernatant. The 64Cu-PTSM labeling procedure did not interfere significantly with C6 cell viability and proliferation rate. MicroPET images of living mice indicate that tail-vein injected labeled C6 cells traffic to the lungs and liver. In addition, transient splenic accumulation of radioactivity was clearly detectable in a mouse scanned at 3.33 h postinfusion of 64Cu-PTSM-labeled lymphocytes. In contrast, the liver was the principal organ of tracer localization after tail-vein administration of 64Cu-PTSM alone. These results indicate that in vivo imaging of cell trafficking is possible with 64Cu-PTSM-labeled cells. Given the longer t(1/2) of 64Cu (12.7 h) relative to 18F (110 min), longer cell-tracking periods (up to 24-36 h) should be possible now with PET. PMID- 11867753 TI - An RBCC protein implicated in maintenance of steady-state neuregulin receptor levels. AB - Despite numerous recent advances in our understanding of the molecular mechanisms underlying receptor tyrosine kinase down-regulation and degradation in response to growth factor binding, relatively little is known about ligand-independent receptor tyrosine kinase degradation mechanisms. In a screen for proteins that might regulate the trafficking or localization of the ErbB3 receptor, we have identified a tripartite or RBCC (RING, B-box, coiled-coil) protein that interacts with the cytoplasmic tail of the receptor in an activation-independent manner. We have named this protein Nrdp1 for neuregulin receptor degradation protein-1. Northern blotting reveals ubiquitous distribution of Nrdp1 in human adult tissues, but message is particularly prominent in heart, brain, and skeletal muscle. Nrdp1 interacts specifically with the neuregulin receptors ErbB3 and ErbB4 and not with epidermal growth factor receptor or ErbB2. When coexpressed in COS7 cells, Nrdp1 mediates the redistribution of ErbB3 from the cell surface to intracellular compartments and induces the suppression of ErbB3 and ErbB4 receptor levels but not epidermal growth factor receptor or ErbB2 levels. A putative dominant-negative form of Nrdp1 potentiates neuregulin-stimulated Erk1/2 activity in transfected MCF7 breast tumor cells. Our observations suggest that Nrdp1 may act to regulate steady-state cell surface neuregulin receptor levels, thereby influencing the efficiency of neuregulin signaling. PMID- 11867754 TI - Genetic instability favoring transversions associated with ErbB2-induced mammary tumorigenesis. AB - It has been argued that genetic instability is required to generate the myriad mutations that fuel tumor initiation and progression and, in fact, patients with heritable cancer susceptibility syndromes harbor defects in specific genes that normally maintain DNA integrity. However, the vast majority of human cancers arise sporadically, in the absence of deficiencies in known "mutator" genes. We used a cII-based mutation detection assay to show that the mean frequency of forward mutations in primary mammary adenocarcinomas arising in mouse mammary tumor virus-c-erbB2 transgenic mice harboring multiple copies of the lambda bacteriophage genome was significantly higher than in aged-matched, wild-type mammary tissue. Analysis of the cII mutational spectrum within the mammary tumor genomic DNA demonstrated a >6-fold elevation in transversion mutation frequency, resulting in a highly unusual inversion of the transition/transversion ratio characteristic of normal epithelium; frameshift mutation frequencies were unaltered. Arising oncogenic point mutations within the c-erbB2 transgene of such tumors were predominantly transversions as well. Data from this model system support the notion that elaboration of a mutator phenotype is a consequential event in breast cancer and suggest that a novel DNA replication/repair gene is a relatively early mutational target in c-erbB2-induced mammary tumorigenesis. PMID- 11867755 TI - Crystal structure and electron transfer kinetics of CueO, a multicopper oxidase required for copper homeostasis in Escherichia coli. AB - CueO (YacK), a multicopper oxidase, is part of the copper-regulatory cue operon in Escherichia coli. The crystal structure of CueO has been determined to 1.4-A resolution by using multiple anomalous dispersion phasing and an automated building procedure that yielded a nearly complete model without manual intervention. This is the highest resolution multicopper oxidase structure yet determined and provides a particularly clear view of the four coppers at the catalytic center. The overall structure is similar to those of laccase and ascorbate oxidase, but contains an extra 42-residue insert in domain 3 that includes 14 methionines, nine of which lie in a helix that covers the entrance to the type I (T1, blue) copper site. The trinuclear copper cluster has a conformation not previously seen: the Cu-O-Cu binuclear species is nearly linear (Cu-O-Cu bond angle = 170 degrees) and the third (type II) copper lies only 3.1 A from the bridging oxygen. CueO activity was maximal at pH 6.5 and in the presence of >100 microM Cu(II). Measurements of intermolecular and intramolecular electron transfer with laser flash photolysis in the absence of Cu(II) show that, in addition to the normal reduction of the T1 copper, which occurs with a slow rate (k = 4 x 10(7) M(-1)x (-1)), a second electron transfer process occurs to an unknown site, possibly the trinuclear cluster, with k = 9 x 10(7) M(-1) x (-1), followed by a slow intramolecular electron transfer to T1 copper (k approximately 10 s(-1)). These results suggest the methionine-rich helix blocks access to the T1 site in the absence of excess copper. PMID- 11867756 TI - Construction of a rationally designed human cytomegalovirus variant encoding a temperature-sensitive immediate-early 2 protein. AB - We generated a set of cysteine-to-glycine mutations and screened them to identify a temperature-sensitive allele of the human cytomegalovirus UL122 gene, which encodes the immediate-early 2 transcriptional activating protein. The mutant allele contains a single base pair substitution at amino acid 510. In transcription activation assays, the mutant protein activated the simian virus 40 early and human cytomegalovirus UL112 promoters at 32.5 degrees C but not at 39.5 degrees C. We constructed a mutant virus, BTNtsUL122, in which the wild-type UL122 locus is substituted with the mutant allele. The mutant produced progeny at 32.5 degrees C but not at 39.5 degrees C. Although the mutant virus accumulated immediate-early transcripts and proteins at the nonpermissive temperature, it did not produce any early (UL44 and UL54) and late (UL82) transcripts and it did not replicate its DNA. The mutant's defect at the nonpermissive temperature results, at least in part, from the inability of the temperature-sensitive immediate-early 2 protein to activate early viral promoters, whose products are required for DNA replication and progression into the late phase of the virus growth cycle. PMID- 11867757 TI - Drug selection with paclitaxel restores expression of linked IL-2 receptor gamma chain and multidrug resistance (MDR1) transgenes in canine bone marrow. AB - Unstable expression of transferred genes is a major obstacle to successful gene therapy of hematopoietic diseases. We have investigated in a canine large-animal model whether expression of transduced genes can be recovered in vivo. Mixed breed dogs had undergone autologous bone marrow transplantation (BMT) with stem cell factor and granulocyte-colony-stimulating factor-mobilized retrovirally marked hematopoietic cells. The bicistronic retroviral vector construct allowed for coexpression of MDR1 and human IL-2 receptor common gamma-chain cDNAs. The latter gene is deficient in X-linked severe combined immunodeficiency. After initial high-level expression, P-glycoprotein and the gamma-chain were undetectable in blood and bone marrow 17 months post-BMT. Six months later, one dog was treated i.v. with 125 mg/m2 paclitaxel. Three administrations restored expression of the two linked genes to high levels in blood and bone marrow. Two dogs treated with higher paclitaxel doses died from myelosuppression after the first administration. As determined by flow cytometry, both genes were expressed in granulocytes, monocytes, and lymphocytes of the surviving animal. PCR analysis of DNA from peripheral blood confirmed that the retroviral cDNA was increased after paclitaxel treatment, suggesting enrichment of transduced cells. P glycoprotein was detectable for more than 1 year after cessation of paclitaxel. Repeated analyses of blood and bone marrow aspirates gave no indication of hematopoietic disturbance after BMT with transduced cells and paclitaxel treatment. In summary, we have shown that with the use of a drug-selectable marker gene, chemotherapy can select for cells that express an otherwise nonselected therapeutic gene in blood and bone marrow. PMID- 11867758 TI - Cybr, a cytokine-inducible protein that binds cytohesin-1 and regulates its activity. AB - Cytokines regulate lymphocyte development and differentiation, but precisely how they control these processes is still poorly understood. By using microarray technology to detect cytokine-induced genes, we identified a cDNA encoding Cybr, which was increased markedly in cells incubated with IL-2 and IL-12. The mRNA was most abundant in hematopoietic cells and tissues. The predicted amino acid sequence is similar to that of GRP-1-associated protein (GRASP), a recently identified retinoic acid-induced cytohesin-binding protein. Physical interaction, dependent on the coiled-coil domains of Cybr and cytohesin-1, was demonstrated by coimmunoprecipitation of the overexpressed proteins from 293T cells. Cytohesin-1, in addition to its role in cell adhesion, is a guanine nucleotide-exchange protein activator of ARF GTPases. Acceleration of guanosine 5prime prime or minute-O-(thiotriphosphate) binding to ARF by cytohesin-1 in vitro was enhanced by Cybr. Because the binding protein modified activation of ADP ribosylation factor by cytohesin-1, we designate this cytokine-inducible protein Cybr (cytohesin binder and regulator). PMID- 11867759 TI - Targeted point mutations of p53 lead to dominant-negative inhibition of wild-type p53 function. AB - The p53 tumor suppressor gene is the most frequently mutated gene in human cancers, and germ-line p53 mutations cause a familial predisposition for cancer. Germ-line or sporadic p53 mutations are usually missense and typically affect the central DNA-binding domain of the protein. Because p53 functions as a tetrameric transcription factor, mutant p53 is thought to inhibit the function of wild-type p53 protein. Here, we studied the possible dominant-negative inhibition of wild type p53 protein by two different, frequently occurring point mutations. The R270H and P275S mutations were targeted into the genome of mouse embryonic stem cells to allow the analysis of the effects of the mutant proteins expressed in normal cells at single-copy levels. In embryonic stem cells, the presence of a heterozygous point-mutated allele resulted in delayed transcriptional activation of several p53 downstream target genes on exposure to gamma irradiation. Doxorubicin-induced apoptosis was severely affected in the mutant embryonic stem cells compared with wild-type cells. Heterozygous mutant thymocytes had a severe defect in p53-dependent apoptotic pathways after treatment with gamma irradiation or doxorubicin, whereas p53-independent apoptotic pathways were intact. Together these data demonstrate that physiological expression of point-mutated p53 can strongly limit overall cellular p53 function, supporting the dominant-negative action of such mutants. Also, cells heterozygous for such mutations may be compromised in terms of tumor suppression and response to chemotherapeutic agents. PMID- 11867760 TI - Protection of cardiac mitochondria by diazoxide and protein kinase C: implications for ischemic preconditioning. AB - Mitochondrial ATP-sensitive K (mitoK(ATP)) channels play a central role in protecting the heart from injury in ischemic preconditioning. In isolated mitochondria exposed to elevated extramitochondrial Ca, P(i), and anoxia to simulate ischemic conditions, the selective mitoK(ATP) channel agonist diazoxide (25-50 microM) potently reduced mitochondrial injury by preventing both the mitochondrial permeability transition (MPT) and cytochrome c loss from the intermembrane space. Both effects were blocked completely by the selective mitoK(ATP) antagonist 5-hydroxydecanoate. The protective effect against Ca induced MPT was most evident under conditions in which the ability of electron transport to support membrane potential (Deltapsi(m)) was decreased and inner membrane leakiness was increased moderately. Under these conditions, mitoK(ATP) channel activity strongly regulated Deltapsi(m), and diazoxide prevented MPT by inhibiting the driving force for Ca uptake. Phorbol 12-myristate 13-acetate mimicked the protective effects of diazoxide, unless 5-hydroxydecanoate was present, indicating that protein kinase C activation also protects mitochondria by activating mitoK(ATP) channels. Because Deltapsi(m) recovery ultimately is required for heart functional recovery, these results may explain how mitoK(ATP) channel activation mimics ischemic preconditioning by protecting mitochondria as they pass through a critical vulnerability window during ischemia/reperfusion. PMID- 11867761 TI - Inhibition of cyclooxygenase 2 blocks human cytomegalovirus replication. AB - Cyclooxygenase 2 (COX-2) mRNA, protein, and activity are transiently induced after infection of human fibroblasts with human cytomegalovirus. Prostaglandin E(2), the product of COX-2 activity, is transiently increased by a factor of >50 in cultures of virus-infected fibroblasts. Both specific (BMS-279652, 279654, and 279655) and nonspecific (indomethacin) COX-2 inhibitors can abrogate the virus mediated induction of prostaglandin E(2) accumulation. Levels of COX-2 inhibitors that completely block the induction of COX-2 activity, but do not compromise cell viability, reduce the yield of human cytomegalovirus in human fibroblasts by a factor of >100. Importantly, the yield of infectious virus can be substantially restored by the addition of prostaglandin E(2) together with the inhibitory drug. This finding argues that elevated levels of prostaglandin E(2) are required for efficient replication of human cytomegalovirus in fibroblasts. COX-2 inhibitors block the accumulation of immediate-early 2 mRNA and protein, but have little effect on the levels of immediate-early 1 mRNA and protein. Viral DNA replication and the accumulation of some, but not all, early and late mRNAs are substantially blocked by COX-2 inhibitors. Elevated levels of prostaglandin E(2) apparently facilitate the production of immediate-early 2 protein. The failure to produce normal levels of this critical viral regulatory protein in the presence of COX-2 inhibitors might block normal progression beyond the immediate-early phase of human cytomegalovirus infection. PMID- 11867763 TI - Identification of a germ-line pro-B cell subset that distinguishes the fetal/neonatal from the adult B cell development pathway. AB - Studies presented here show that the expression of CD4, MHC class II (Ia,) and B220 cleanly resolves a major and a minor subset within the earliest pro-B cell population (germ-line pro-B) in adult bone marrow (BM). The major subset expresses intermediate B220 and low CD4 levels. The minor subset, which constitutes roughly 20% of the adult germ-line pro-B, expresses very low B220 levels and does not express CD4. Ia is clearly detectable at low levels on the major germ-line pro-B subset, both in wild-type adult mice and in gene-targeted mice (RAG2-/- and microMT), in which B cell development terminates before the pre B cell stage. A small proportion of cells in the more mature pro-B cell subsets (Hardy Fractions B and C) also express Ia at this level. In contrast, Ia levels on the minor subset are barely above (or equal to) background. Surprisingly, the major germ-line pro-B cell subset found in adults is missing in fetal and neonatal animals. All of the germ-line pro-B in these immature animals express a phenotype (very low B220, no CD4, or Ia) similar to that of the minor pro-B cell subset in adult BM. Because B cell development in fetal/neonatal animals principally results in B-1 cells, these findings demonstrate that the B-1 development pathway does not include the major germ-line pro-B subset found in adult BM and hence identify a very early difference between the B-1 and -2 development pathways. PMID- 11867764 TI - Conserved tryptophan in the core domain of transglutaminase is essential for catalytic activity. AB - Transglutaminase 2 (TG2) is a distinctive member of the family of Ca2+-dependent enzymes recognized mostly by their abilities to catalyze the posttranslational crosslinking of proteins. TG2 uniquely binds and hydrolyzes GTP; binding GTP inhibits its crosslinking activity but allows it to function in signal transduction (hence the G(h) designation). The core domain of TG2 (residues 139 471, rat) comprises the papain-like catalytic triad and the GTP-binding domain (residues 159-173) and contains almost all of the conserved tryptophans of the protein. Examining point mutations at Trp positions 180, 241, 278, 332, and 337 showed that, upon binding 2'-(or 3')-O-(N-methylanthraniloyl)GTP (mantGTP), the Phe-332 mutant was the weakest (35% less than wild type) in resonance energy transfer from the protein (lambda(exc, max) = 290 nm) to the mant fluorophore (lambda(em) = 444 nm) and had a reduced affinity for mantGTP. Trp-332, situated near the catalytic center and the nucleotide-binding area of TG2, may be part of the allosteric relay machinery that transmits negative effector signals from nucleotide binding to the active center of TG2. A most important observation was that, whereas no enzyme activity could be detected when Trp-241 was replaced with Ala or Gln, partial preservation of catalytic activity was seen with substitutions by Tyr > Phe > His. The results indicate that Trp-241 is essential for catalysis, possibly by stabilizing the transition states by H-bonding, quadrupole-ion, or van der Waals interactions. This contrasts with the evolutionarily related papain family of cysteine proteases, which uses Gln-19 (papain) for stabilizing the transition state. PMID- 11867762 TI - Interferon regulatory factor-3 is an in vivo target of DNA-PK. AB - Eukaryotic cells have evolved complex signaling networks to sense environmental stress and to repair stress-induced damage. IFN regulatory factor-3 (IRF-3) is a transcription factor that plays a central role in the host response to viral infection. Although the main activity of IRF-3 characterized to date has been its role in the induction of IFN-alpha and -beta after virus infection, recent evidence indicates additional roles for IRF-3 in the response to DNA damage and in virus-induced apoptosis. Here we identify IRF-3 as the first in vivo target for DNA-dependent protein kinase (DNA-PK). Phosphorylation of IRF-3 by DNA-PK after virus infection results in its nuclear retention and delayed proteolysis. These results expand the known roles of DNA-PK and provide a functional link between the cellular machineries that regulate the innate immune response and that sense and respond to DNA damage. As such this study contributes to a more integrated view of the cellular responses to various cellular stress signals. PMID- 11867765 TI - Analysis of the AAA sensor-2 motif in the C-terminal ATPase domain of Hsp104 with a site-specific fluorescent probe of nucleotide binding. AB - Hsp104 from Saccharomyces cerevisiae is a hexameric protein with two AAA ATPase domains (N- and C-terminal nucleotide-binding domains NBD1 and NBD2, respectively) per monomer. Our previous analysis of the Hsp104 ATP hydrolysis cycle revealed that NBD1 and NBD2 have very different catalytic properties, but each shows positive cooperativity in hydrolysis. There is also communication between the two domains, in that ATP hydrolysis at NBD1 depends on the nucleotide that is bound to NBD2. Here, we extend our understanding of the Hsp104 ATP hydrolysis cycle through mutagenesis of the AAA sensor-2 motif in NBD2. To do so, we took advantage of the lack of tryptophan residues in Hsp104 to place a single tryptophan in the C-terminal domain (Y819W). The Y819W substitution has no significant effects on folding stability of the C-terminal domain or on ATP hydrolysis by NBD1 or NBD2. The fluorescence of this tryptophan changes in response to ATP and ADP binding, allowing the K(d) and Hill coefficient to be determined for each nucleotide. By using this site-specific probe of binding, we analyze the effect of mutating the conserved arginine residue in the sensor-2 motif in Hsp104 NBD2. An R826M mutation causes nearly equal decreases in affinity of NBD2 for both ATP and ADP, indicating that at this site, the sensor-2 provides binding energy, but does not act to sense the difference between these nucleotides. In addition, the rate of ATP hydrolysis at NBD1 is decreased by the R826M mutation, providing further evidence for interdomain communication in the Hsp104 ATP hydrolysis cycle. PMID- 11867767 TI - Prevalence of somatic alterations in the colorectal cancer cell genome. AB - Although a small fraction of human cancers have increased rates of somatic mutation because of known deficiencies in DNA repair, little is known about the prevalence of somatic alterations in the vast majority of human cancers. To systematically assess nonsynonymous somatic alterations in colorectal neoplasia, we used DNA sequencing to analyze approximately 3.2 Mb of coding tumor DNA comprising 1,811 exons from 470 genes. In total, we identified only three distinct somatic mutations, comprising two missense changes and one 14-bp deletion, each in a different gene. The accumulation of approximately one nonsynonymous somatic change per Mb of tumor DNA is consistent with a rate of mutation in tumor cells that is similar to that of normal cells. These data suggest that most sporadic colorectal cancers do not display a mutator phenotype at the nucleotide level. They also have significant implications for the interpretation of somatic mutations in candidate tumor-suppressor genes. PMID- 11867766 TI - Neuronal nitric-oxide synthase localization mediated by a ternary complex with synapsin and CAPON. AB - The specificity of the reactions of nitric oxide (NO) with its neuronal targets is determined in part by the precise localizations of neuronal NO synthase (nNOS) within the cell. The targeting of nNOS is mediated by adapter proteins that interact with its PDZ domain. Here, we show that the nNOS adapter protein, CAPON, interacts with synapsins I, II, and III through an N-terminal phosphotyrosine binding domain interaction, which leads to a ternary complex comprising nNOS, CAPON, and synapsin I. The significance of this ternary complex is demonstrated by changes in subcellular localization of nNOS in mice harboring genomic deletions of both synapsin I and synapsin II. These results suggest a mechanism for specific actions of NO at presynaptic sites. PMID- 11867768 TI - Phosphatidylinositol 4,5-bisphosphate stimulates vesicle formation from liposomes by brain cytosol. AB - As a step toward the elucidation of mechanisms in vesicle budding, a cell-free assay that measures cytosol-induced vesicle generation from liposomes was established. This assay then was used to explore the role of phosphoinositides in vesicle formation. Liposomes incubated with brain cytosol in the presence of ATP and GTP massively generated small vesicles, as assessed both quantitatively and qualitatively by a dynamic light-scattering assay. Both ATP and GTP were required. Vesicle formation was inhibited greatly by the immunodepletion of dynamin 1 from the cytosol, indicating a major contribution of this GTPase in this reaction and suggesting that it mimics endocytic vesicle fission. Increasing the concentration of l-alpha-phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P2] but not of l-alpha-phosphatidylinositol 4-monophosphate or l alpha-phosphatidylinositol in the lipid membranes enhanced vesicle formation. Lipid analysis revealed rapid degradation of PtdIns(4,5)P2 to l-alpha phosphatidylinositol during the incubation with the reaction reaching a maximum within 5 sec, whereas vesicle formation proceeded with a longer time course. PtdIns(4,5)P2 degradation was independent of vesicle formation and occurred also in the presence of guanosine 5'-O-(thiotriphosphate), where few vesicle formations occurred. These results suggest that PtdIns(4,5)P2 plays a critical role in the early step of vesicle formation, possibly in the recruitment of coats and fission factors to membranes. PMID- 11867770 TI - Study design for the new millennium: changing how we perform research and practice medicine. PMID- 11867769 TI - The TRAP/Mediator coactivator complex interacts directly with estrogen receptors alpha and beta through the TRAP220 subunit and directly enhances estrogen receptor function in vitro. AB - Target gene activation by nuclear hormone receptors, including estrogen receptors (ERs), is thought to be mediated by a variety of interacting cofactors. Here we identify a number of nuclear extract-derived proteins that interact with immobilized ER ligand binding domains in a 17beta-estradiol-dependent manner. The most prominent of these are components of the thyroid hormone receptor-associated protein (TRAP)/Mediator coactivator complex, which interacts with ERalpha and ERbeta in both unfractionated nuclear extracts and purified form. Studies with extracts from TRAP220(-/-) fibroblasts reveal that these interactions depend on TRAP220, a TRAP/Mediator subunit previously shown to interact with ER and other nuclear receptors in a ligand-dependent manner. The physiological relevance of the in vitro interaction is documented further by the isolation of an ERalpha TRAP/Mediator complex from cultured cells expressing an epitope-tagged ERalpha. Finally, the complete TRAP/Mediator complex is shown to enhance ER function directly in a highly purified cell-free transcription system. These studies firmly establish a direct role for TRAP/Mediator, through TRAP220, in ER function. PMID- 11867771 TI - US for postmenopausal bleeding: consensus development and patient-centered outcomes. PMID- 11867772 TI - Teaching the teachers. PMID- 11867773 TI - Study design for concurrent development, assessment, and implementation of new diagnostic imaging technology. AB - With current constraints on health care resources and emphasis on value for money, new diagnostic imaging technologies must be assessed and their value demonstrated. The state of the art in the field of diagnostic imaging technology assessment advocates a hierarchical step-by-step approach. Although rigorous, such a hierarchical assessment is time-consuming, and, given the current rapid advances in technology, results are often too late to influence management and policy decisions. The purpose of this article is to discuss a study design in which development, assessment, and implementation of new diagnostic imaging technology take place concurrently in one integrated process. An empirically based pragmatic study design is proposed for imaging technology assessment. To minimize bias and enable comparison with current technology, a randomized controlled design is used whenever feasible and ethical. Outcome measures should reflect the clinical decision-making process based on imaging information and acceptance of the new test. Outcome measures can include additional imaging studies requested, costs of diagnostic work-up and treatment, physicians' confidence in therapeutic decision making, recruitment rate, and patient outcome measures related to the clinical problem. The key feature of the proposed study design is analysis of trends in outcome measures over time. PMID- 11867774 TI - Trends in the utilization of MR angiography and body MR imaging in the US Medicare population: 1993-1998. AB - PURPOSE: To determine the trends in utilization of non-neurologic (ie, body) magnetic resonance (MR) imaging and of MR angiographic examinations performed from 1993 to 1998, the trends in non-radiologist participation in MR imaging, and the relative reimbursements for these examinations compared with those for all other noninvasive imaging studies performed in 1998. MATERIALS AND METHODS: By using the 1993, 1996, and 1998 nationwide Medicare Part B databases, utilization rates per 100,000 Medicare beneficiaries and physician reimbursements were determined for seven MR angiography and 14 body MR imaging CPT-4 (Current Procedural Terminology, version 4) codes. Medicare specialty codes were used to categorize physicians as radiologists or non-radiologists. RESULTS: The utilization rate per 100,000 Medicare beneficiaries for all 21 MR angiography and body MR imaging codes increased from 649 in 1993 to 1,253 in 1996 and to 1,876 in 1998--a 189% increase. These rates represented 0.55% of the total noninvasive imaging volume in 1998 and 2.8% of physician reimbursements. Musculoskeletal MR imaging utilization increased 142% from 1993 to 1998 compared with a 58% increase in the utilization of other body MR imaging studies. Non-radiologist participation in musculoskeletal MR imaging increased from 2.9% in 1993 to 3.6% in 1996 and to 5.6% in 1998. CONCLUSION: MR angiography and body MR imaging utilization rates increased substantially from 1993 to 1998. However, these studies still account for a minor fraction of all noninvasive imaging examinations performed and fees reimbursed. MR angiography and musculoskeletal MR imaging utilization has increased rapidly. Non-radiologist participation in musculoskeletal MR imaging is increasing. PMID- 11867777 TI - Adrenal masses: characterization with combined unenhanced and delayed enhanced CT. AB - PURPOSE: To assess the accuracy of a dedicated adrenal computed tomographic (CT) protocol. MATERIALS AND METHODS: One hundred sixty-six adrenal masses were evaluated with a protocol consisting of unenhanced CT, and, for those with attenuation values greater than 10 HU, contrast material-enhanced and delayed enhanced CT. Attenuation values and enhancement washout calculations were obtained. An adenoma was diagnosed if a mass had an attenuation value of 10 HU or less at unenhanced CT or a percentage enhancement washout value of 60% or higher. RESULTS: The final diagnosis was adenoma in 127 masses and non-adenoma in 39. Masses measuring more than 10 HU on unenhanced CT scans were confirmed at biopsy (n = 28) or were examined for stability or change in size at follow-up CT performed at a minimum interval of 6 months (n = 33). Thirty-six (92%) of 39 non adenomas and 124 (98%) of 127 adenomas were correctly characterized. The sensitivity and specificity of this protocol were 98% and 92%, respectively. This protocol correctly characterized 160 (96%) of 166 masses. CONCLUSION: With a combination of unenhanced and delayed enhanced CT, nearly all adrenal masses can be correctly categorized as adenomas or non-adenomas. PMID- 11867775 TI - Cost analysis model: US versus endometrial biopsy in evaluation of peri- and postmenopausal abnormal vaginal bleeding. AB - PURPOSE: To develop a cost minimization analysis model from the societal perspective of Medicare reimbursement to determine whether endometrial biopsy or transvaginal ultrasonography (US) is less expensive in evaluating peri- and postmenopausal women with abnormal vaginal bleeding and to assess whether this strategy is equally effective in populations at low and high risk for endometrial carcinoma. MATERIALS AND METHODS: Clinical algorithms were constructed that detailed diagnostic evaluation of the target population by using office-based endometrial biopsy versus transvaginal US as starting points. An economic model based on Medicare reimbursement and average wholesale drug price data and using disease prevalences and modality sensitivities from the scientific literature was then created to examine common bleeding causes in this population. All models included the cost of obtaining a tissue diagnosis for focal or diffuse endometrial thickening found at US. Modality sensitivities and prevalences of disease states were varied within the model to discover limits at which each modality became cheaper versus the other for assessing a population of women. RESULTS: Population prevalence of neoplastic disease is the principal factor governing total cost between competing diagnostic algorithms. In populations with 31% or less combined prevalence of endometrial carcinoma/atypical adenomatous hyperplasia, algorithms utilizing transvaginal US as the initial test are most cost minimizing. At combined endometrial carcinoma/atypical adenomatous hyperplasia prevalence of 10%, savings of up to 11% and 16% over pathways initiated with endometrial biopsy are predicted. In populations with a high incidence of neoplastic disease (>31%), biopsy-based algorithms should become least costly. CONCLUSION: Transvaginal US-initiated triage predicts substantial cost savings versus biopsy-based algorithms in evaluating typical populations of peri- and postmenopausal women with abnormal vaginal bleeding seen in clinical practice. PMID- 11867778 TI - High-frequency Doppler US of the prostate: effect of patient position. AB - PURPOSE: To evaluate cancer detection with directed biopsy of the prostate on the basis of high-frequency Doppler ultrasonographic (US) findings, and to determine the effect of patient position on the observed flow pattern. MATERIALS AND METHODS: Thirty-two patients were evaluated in the left lateral decubitus position with gray-scale, color Doppler, and power Doppler transrectal US. Up to four directed biopsy specimens were obtained on the basis of gray-scale and Doppler US findings, and modified sextant biopsy followed. Analysis of variance and the Wilcoxon signed rank test were used to evaluate the distribution of Doppler signals within the prostate. Three healthy volunteers with no known prostate disease were also examined in supine and both decubitus positions. RESULTS: In the patient group, both color and power Doppler US demonstrated increased flow on the left side of the prostate, with greater flow toward the base of the gland (P <.002). Consequently, 62 of 90 directed-biopsy cores were obtained in the left base and mid-gland. The positive biopsy rate for directed biopsy was not significantly different from that of sextant biopsy (P =.4). Seven patients had cancer that was identified with sextant biopsy, but only four cancers were identified with directed biopsy. Each of the three healthy volunteers demonstrated increased Doppler flow on the dependent side when the subject was in the lateral decubitus position. CONCLUSION: The positive yield of directed biopsy was similar to the yield of sextant biopsy. On the basis of observations made in healthy volunteers, the authors conclude that flow asymmetry in patients who underwent biopsy may have been related to patient position. PMID- 11867779 TI - Embolization of bleeding residual uterine vascular malformations in patients with treated gestational trophoblastic tumors. AB - PURPOSE: To retrospectively evaluate embolotherapy of bleeding residual uterine vascular malformations in patients with gestational trophoblastic tumors. MATERIALS AND METHODS: Fourteen patients were treated over the past 20 years. Embolizations were performed with a common femoral artery approach. Duplex ultrasonography was performed before and after embolization to document the uterine vascularity. The technique and materials used for each embolization, control of hemorrhage, need for repeat embolization, complications, and outcome of subsequent pregnancies were assessed. RESULTS: Hemorrhage was controlled in 11 of the 14 patients; two patients required hysterectomy and one required uterine artery ligation for failure to control hemorrhage after initial embolization. Six patients required repeat embolization for recurrence of bleeding. Therapeutic benefit and success were associated with the ability to selectively embolize the uterine artery and to achieve a greater than 80% reduction in vascular malformation size. Pulsatility indexes of the uterine arteries and endometrial encroachment were not predictive of recurrent hemorrhage. Two patients delivered a total of three full-term infants, one patient experienced a miscarriage, and another experienced a termination of pregnancy following embolotherapy. Pain requiring opiate analgesia was a frequent complication of treatment. CONCLUSION: Selective uterine artery embolization is a safe and effective treatment for severe bleeding from residual uterine vascular malformations in patients with treated gestational trophoblastic tumors. PMID- 11867780 TI - Percutaneous abscess drainage in Crohn disease: technical success and short- and long-term outcomes during 14 years. AB - PURPOSE: To determine technical success with percutaneous abscess drainage (PAD) in patients with Crohn disease during 14 years. MATERIALS AND METHODS: Medical records of 32 patients with Crohn disease who underwent PAD from 1985 to 1999 were reviewed. Results of abscess drainage and nature of subsequent surgical procedures were recorded. Factors assessed included postoperative or spontaneous nature of the abscess, documentation of a proved fistula, history of occurrence of prior abscesses, duration of Crohn disease, and use of steroid treatment. Technical success was defined as complete abscess drainage. Short-term success was defined as avoidance of surgery within 60 days of drainage. Long-term success was defined as avoidance of surgery beyond the initial 60-day period. Short-term avoidance of surgery was assessed as a predictor of the need for surgery in the long term. Statistical analysis was performed with the chi(2) test to evaluate predictors of short-term success and to assess short-term success as a predictor of long-term success. RESULTS: The technical success rate was 96%. In 16 (50%) of 32 patients, the need for surgery in the short term was avoided, and surgery was more likely to be avoided in patients with postoperative abscesses than in those with spontaneous abscesses (P =.07). At long-term follow-up, short-term avoidance of surgery did not significantly increase the likelihood of need for surgery in the long term, which occurred in nine of 16 short-term successes versus five of 15 short-term failures (P =.55). Recurrent abscesses occurred in seven (22%) patients, a rate comparable to that with surgical abscess drainage; four (44%) of nine cases of re-drainage were successful. CONCLUSION: PAD has a high technical success rate of 96%. Half of patients may avoid surgery in the short term. PMID- 11867781 TI - Crohn disease with endoscopic correlation: single-shot fast spin-echo and gadolinium-enhanced fat-suppressed spoiled gradient-echo MR imaging. AB - PURPOSE: To compare T2-weighted breath-hold single-shot fast spin-echo (SE) and gadolinium-enhanced spoiled gradient-echo (GRE) MR imaging with contrast material administered orally and rectally for evaluating patients with Crohn disease. MATERIALS AND METHODS: Twenty-eight patients with Crohn disease received 2% barium sulfate and water enema. The abdomen and pelvis were imaged with transverse and coronal single-shot fast SE and gadolinium-enhanced spoiled GRE MR imaging. Two radiologists reviewed the two types of images for bowel disease. The extent, severity, and conspicuity of the disease were determined. Proof of bowel disease at MR imaging was compared with that at endoscopy, barium study, and surgery. Statistical analysis was performed with the McNemar test. RESULTS: Twenty-five of 28 patients had proven abnormal bowel segments. The per-patient sensitivity of gadolinium-enhanced spoiled GRE MR imaging for the two radiologists was 100% and 96% versus 60% and 60% (P <.05) with single-shot fast SE MR imaging. Gadolinium-enhanced spoiled GRE MR images depicted more segments (54 and 52 of 61 segments; sensitivity, 89% and 85%, respectively) of the diseased bowel than did single-shot fast SE MR images (31 and 32 of 61 segments; sensitivity, 51% and 52%, respectively; P <.001). Severity of Crohn disease was correctly depicted at gadolinium-enhanced spoiled GRE imaging in 93% of patients versus in 43% of patients at single-shot fast SE imaging. CONCLUSION: In patients with Crohn disease, gadolinium-enhanced fat-suppressed spoiled GRE MR imaging better depicted the extent and severity of intestinal disease compared with single-shot fast SE imaging. PMID- 11867782 TI - Superparamagnetic iron oxide-mediated hepatic signal intensity change in patients with and without cirrhosis: pulse sequence effects and Kupffer cell function. AB - PURPOSE: To analyze superparamagnetic iron oxide (SPIO)-mediated hepatic signal intensity change in cirrhotic and noncirrhotic liver and to investigate the relationship between pulse sequence effects in SPIO-enhanced magnetic resonance (MR) imaging for hepatic cirrhosis. MATERIALS AND METHODS: Twelve patients with and 12 patients without cirrhosis underwent T2-weighted fast spin-echo, T2* weighted gradient-echo (GRE), and T1-weighted GRE MR imaging before and twice (early and late phase) after SPIO administration. To assess the effect of SPIO, postcontrast relative signal-to-noise ratio (SNR) changes were statistically analyzed with repeated measurements analysis of variance for each pulse sequence. RESULTS: No interaction was shown between groups and data time points for any pulse sequence. There was no significant difference in mean hepatic relative SNR change on T2-weighted fast spin-echo images between the cirrhotic group and noncirrhotic group (-38.6% and -40.7%, early phase; -42.2% and -49.6%, late phase, respectively). For GRE images, statistically significant differences in mean hepatic relative SNR change were found between the cirrhotic group and noncirrhotic group (-14.2% and -44.5%, early phase; -28.5% and -56.4%, late phase on T2*-weighted GRE images (P <.001); 31.8% and 12.9%, early phase; 23.8% and 2.2%, late phase on T1-weighted GRE images (P <.05), respectively. CONCLUSION: Decreased overall phagocytic activity in cirrhotic liver is more likely due to Kupffer cell dysfunction than to Kupffer cell depletion, since magnetic susceptibility effects on T2*-weighted GRE images depend on intracellular SPIO cluster size. PMID- 11867783 TI - Primary hepatic angiosarcoma: findings at CT and MR imaging. AB - PURPOSE: To evaluate and describe cross-sectional imaging findings in patients with pathologically confirmed primary hepatic angiosarcoma. MATERIALS AND METHODS: Findings from imaging examinations in 13 patients with pathologically confirmed primary hepatic angiosarcoma were retrospectively reviewed (computed tomographic [CT] images obtained in 10 patients and magnetic resonance [MR] images obtained in five patients were available for review). Two gastrointestinal radiologists evaluated lesion number, size, attenuation and signal intensity characteristics, and the pattern and degree of contrast material enhancement. Medical records were reviewed for clinical features associated with angiosarcoma. RESULTS: Angiosarcoma appeared as multiple nodules (n = 6), as dominant masses (n = 6), or as a diffusely infiltrating lesion (n = 1). Multiple nodules were hypoattenuating at unenhanced and contrast material--enhanced CT (six of six patients). When dominant masses were encountered at MR imaging, T2-weighted MR imaging demonstrated heterogeneous internal architecture (four of four patients) similar to that of hepatocellular carcinoma. Multiphase contrast-enhanced CT and MR images showed dominant masses to have heterogeneous and progressive enhancement (three of three patients). Clinical features associated with angiosarcoma included splenic metastases (six of 13 patients), thrombocytopenia (seven of 13 patients), disseminated intravascular coagulation (four of 13 patients), and hemolytic anemia (three of 13 patients). CONCLUSION: Primary hepatic angiosarcoma exhibits a spectrum of appearances that reflect its varied pathologic features. PMID- 11867784 TI - Pancreatic duct: morphologic evaluation with MR cholangiopancreatography after secretin stimulation. AB - PURPOSE: To evaluate the pancreatic duct after administration of secretin. MATERIALS AND METHODS: Single-shot turbo spin-echo T2-weighted dynamic magnetic resonance cholangiopancreatography (MRCP) was performed in 85 patients who did not have pancreatic diseases (group 1) and in 50 patients who had focal severe stenosis of the main pancreatic duct (group 2). The visualization and diameter of the pancreatic duct before and after secretin administration were assessed. RESULTS: In group 1, after secretin administration, the best visualization of the main pancreatic duct in the head, body, accessory pancreatic duct, and branch ducts was achieved in 4.7 minutes +/- 1.6 (SD), 4.8 minutes +/- 1.6, 4.6 minutes +/- 1.6, and 4.7 minutes plus minus 1.3, respectively. Improvement in the delineation of the main pancreatic duct in the head (78 [92%] patients), body (80 [94%] patients), accessory pancreatic duct (35 [41%] patients), and branch ducts (14 [16%] patients) was achieved. Overlap of fluid in the organ and the pancreatic duct was observed in 20 (24%) of 85 patients. Overlap was especially observed after 5 minutes of secretin injection. In group 2, the best visualization of the distal main pancreatic duct was achieved 4.9 minutes +/- 1.4 after secretin administration. Improvement in the delineation of the distal main pancreatic duct was achieved in 17 (85%) of 20 patients. CONCLUSION: MRCP is best performed during the first 5 minutes after secretin administration. PMID- 11867785 TI - Conformal therapy for pancreatic cancer: variation of organ position due to gastrointestinal distention--implications for treatment planning. AB - PURPOSE: To quantify nonrespiratory organ motion in the pancreatic region and its effect on clinical target volume. MATERIALS AND METHODS: Three-dimensional translations of the geometric centers of the volumes of interest--pancreatic head, body, and tail; left and right kidney; and the superior mesenteric artery- were measured in 20 patients by analyzing three spiral computed tomographic (CT) protocols performed at static exhalation and representing differential gastrointestinal distention. Wilcoxon test for paired differences was applied to determine statistical significance (P <.05). Spearman rank correlation coefficients were calculated between combinations of statistically significant translations. With the assumption that the organ positions were represented by a three-dimensional Gaussian distribution that occurs during treatment, clinical target volume expansions were calculated to account for organ motion and a typical setup error. RESULTS: Significant translations of the volume of interest were observed. The most mobile parts of the target organs were the pancreatic tail (P =.001) and the superior mesenteric artery (P =.01). Larger variations from the mean in the planning CT protocol in which negative contrast material was used usually resulted in a slightly larger clinical target volume expansion. CONCLUSION: Our data may provide a basis for further studies of organ motion and ways of modifying treatment margins. PMID- 11867786 TI - MR cholangiography in the evaluation of neonatal cholestasis: initial results. AB - PURPOSE: To retrospectively analyze prospective magnetic resonance (MR) cholangiographic interpretations of findings and compare them with clinical outcome and to determine the accuracy of MR cholangiography in depicting extrahepatic biliary atresia and helping to distinguish it from other causes of neonatal jaundice. MATERIALS AND METHODS: Twenty-six infants (15 male, 11 female; median age, 2 months) underwent MR cholangiography with a 1.5-T MR imaging unit. Original interpretations were compared with clinical outcome. Statistical analysis was performed to determine the accuracy of MR cholangiography in depicting extrahepatic biliary atresia. Equivocal cases and any cases lost to follow-up were excluded. RESULTS: Findings in six of 26 infants were interpreted as normal, and none of five patients (one lost to follow-up) had biliary atresia or other surgical lesions; two were abnormal but not suggestive of biliary atresia (one false-negative finding); 12 were consistent with biliary atresia (three false-positive findings); four demonstrated a choledochal cyst; and two were equivocal. MR cholangiography accuracy was 82% (19 of 23); sensitivity, 90% (nine of 10); and specificity, 77% (10 of 13) for the detection of extrahepatic biliary atresia, with a positive predictive value of 75% (nine of 12) and a negative predictive value of 91% (10 of 11). CONCLUSION: Results of this study found that MR cholangiography is 82% accurate, 90% sensitive, and 77% specific for depicting extrahepatic biliary atresia. Contrary to previous reports, false positive and false-negative findings occur at MR cholangiography. PMID- 11867787 TI - Angiographic classification of hepatic hemangiomas in infants. AB - PURPOSE: To review the angiograms in patients with hepatic hemangiomas referred to two North American children's hospitals to determine the variability in angiographic findings and to propose a classification system that is based on these findings. MATERIALS AND METHODS: Angiograms obtained in 15 infants with a diagnosis of hepatic hemangioma who were examined at or referred to two tertiary pediatric hospitals in North America from 1981 through 2000 were reviewed. The angiographic findings were then used to classify hemangiomas into types on the basis of a number of features, including high-flow nodules, early filling of veins, and the type of direct shunt present. Clinical data, including age at presentation, presence of cardiac insufficiency, and treatment, were also recorded. RESULTS: Lesions were classified into five types on the basis of angiographic findings. In three of 15 patients, angiograms demonstrated the classic appearance of hepatic hemangiomas, with early filling of abnormal vascular channels, stagnation of contrast material, and no evidence of a direct shunt (type 1). In four patients, images showed high-flow nodules without direct shunts (type 2). In eight patients, direct shunts were demonstrated: arteriovenous shunts (type 3) in one, portovenous shunts (type 4) in three, and both arteriovenous and portovenous shunts (type 5) in four. CONCLUSION: Hepatic hemangioma in infants is a heterogeneous lesion with variable angioarchitecture and a spectrum of angiographic findings. PMID- 11867788 TI - Posttransplantation lymphoproliferative disorder: manifestations in pediatric thoracic organ recipients. AB - PURPOSE: To describe and correlate the clinical and imaging features of posttransplantation lymphoproliferative disorder (PTLD) in young thoracic organ transplant recipients. MATERIALS AND METHODS: The authors retrospectively reviewed the medical and imaging records of 31 PTLD episodes in 27 patients with PTLD out of 183 young patients who survived for at least 1 month after thoracic organ transplantation: 18 (14%) heart transplant recipients and nine (16%) lung or heart-lung transplant recipients. Four patients had two separate PTLD episodes. The distribution, timing, and imaging features of the disease were analyzed. RESULTS: Seventeen (55%) of 31 episodes involved intrathoracic PTLD manifesting as multiple pulmonary nodules (n = 10), a solitary nodule (n = 3), alveolar consolidation (n = 3), and/or mediastinal adenopathy (n = 8). Extrathoracic PTLD occurred in 21 (68%) of 31 episodes and involved the abdomen (n = 15), head and neck (n = 11), and/or central nervous system (n = 3). The imaging findings of these episodes included bowel wall thickening, lymphadenopathy, and focal masses. Intrathoracic PTLD occurred more commonly in lung transplant recipients (89%) than in heart transplant recipients (44%); no cases of lymphoma involved the thorax. The frequency of extrathoracic manifestations was higher in heart transplant recipients (83%) than in lung transplant recipients (33%). In lung transplant recipients, the prevalence of early-onset PTLD was significantly greater than that in heart transplant recipients (P <.05). Intrathoracic PTLD tended to manifest early. CONCLUSION: PTLD in young thoracic transplant recipients involves the lungs and extrathoracic organs, tends to have an early onset, and manifests predominantly in the thorax in lung transplant and heart-lung transplant recipients, as opposed to heart transplant recipients. PMID- 11867789 TI - Screening for stroke in sickle cell anemia: comparison of transcranial Doppler imaging and nonimaging US techniques. AB - PURPOSE: To determine whether criteria for screening patients with sickle cell anemia for stroke established with a nonimaging transcranial Doppler ultrasonographic (US) technique are applicable to studies performed with a transcranial Doppler US imaging technique. MATERIALS AND METHODS: One hundred sixty-eight examinations in 66 children were performed for sickle cell stroke screening. Children were examined with nonimaging and imaging transcranial Doppler US techniques on the same day, for a total of 84 paired examinations. The time-averaged maximum mean velocity (V(mean)) and resistive index (RI) were calculated in the middle cerebral arteries, bifurcations of the distal internal carotid arteries, distal internal carotid arteries, anterior cerebral arteries, posterior cerebral arteries, and basilar arteries. The maximum systolic velocity (V(max)) was evaluated in the distal internal carotid arteries and middle cerebral arteries. V(mean), V(max), and RI measurements were subjected to repeated-measures multivariate analysis of covariance, and the Pearson product moment correlation was used for middle cerebral artery velocity, age, and hemoglobin. RESULTS: V(mean) measurements obtained with nonimaging and imaging techniques varied substantially for the bifurcation of the distal internal carotid artery, the posterior cerebral artery, and the basilar artery. Substantial differences were found in RIs for every vessel. Examination time was shorter with the nonimaging technique. CONCLUSION: V(mean) measurements in the middle cerebral artery, distal internal carotid artery, and anterior cerebral artery did not vary substantially between nonimaging and imaging transcranial Doppler US. RI data did not yield comparable measurements. PMID- 11867790 TI - High-grade gliomas and solitary metastases: differentiation by using perfusion and proton spectroscopic MR imaging. AB - PURPOSE: To determine whether perfusion-weighted and proton spectroscopic MR imaging can be used to differentiate high-grade primary gliomas and solitary metastases on the basis of differences in vascularity and metabolite levels in the peritumoral region. MATERIALS AND METHODS: Fifty-one patients with a solitary brain tumor (33 gliomas, 18 metastases) underwent conventional, contrast material -enhanced perfusion-weighted, and proton spectroscopic MR imaging before surgical resection or stereotactic biopsy. Of the 33 patients with gliomas, 22 underwent perfusion-weighted MR imaging; nine, spectroscopic MR imaging; and two underwent both. Of the 18 patients with metastases, 12 underwent perfusion-weighted MR imaging, and six, spectroscopic MR imaging. The peritumoral region was defined as the area in the white matter immediately adjacent to the enhancing (hyperintense on T2-weighted images, but not enhancing on postcontrast T1-weighted images) portion of the tumor. Relative cerebral blood volumes in these regions were calculated from perfusion-weighted MR data. Spectra from the enhancing tumor, the peritumoral region, and normal brain were obtained from the two-dimensional spectroscopic MR acquisition. The Student t test was used to determine if there was a statistically significant difference in relative cerebral blood volume and metabolic ratios between high-grade gliomas and metastases. RESULTS: The measured relative cerebral blood volumes in the peritumoral region in high-grade gliomas and metastases were 1.31 +/- 0.97 (mean +/- SD) and 0.39 +/- 0.19, respectively. The difference was statistically significant (P <.001). Spectroscopic imaging demonstrated elevated choline levels (choline-to-creatine ratio was 2.28 +/- 1.24) in the peritumoral region of gliomas but not in metastases (choline-to creatine ratio was 0.76 +/- 0.23). The difference was statistically significant (P =.001). CONCLUSION: Although conventional MR imaging characteristics of solitary metastases and primary high-grade gliomas may sometimes be similar, perfusion-weighted and spectroscopic MR imaging enable distinction between the two. PMID- 11867791 TI - Systemic lupus erythematosus: diagnostic application of magnetization transfer ratio histograms in patients with neuropsychiatric symptoms--initial results. AB - PURPOSE: To explore the diagnostic potential of magnetization transfer ratio (MTR) histogram analysis in patients with neuropsychiatric systemic lupus erythematosus (SLE) by using multivariate discriminant analysis (MDA). MATERIALS AND METHODS: Volumetric magnetization transfer imaging was performed in nine patients with active non-thromboembolic, neuropsychiatric SLE, 10 patients with SLE who had had neuropsychiatric SLE previously, 10 patients with SLE but no history of neuropsychiatric SLE, 10 patients with inactive multiple sclerosis, and 10 healthy control subjects. For each subject, an MTR histogram of the whole brain was generated, and an MDA score was produced for each histogram. Each patient was assigned to a clinical subgroup on the basis of these MDA scores. For assignment, binary comparisons between subgroups were made. The accuracy of this classification method was assessed and compared with that of conventional MTR histogram analysis. RESULTS: With MDA, the success rate of binary classification was 60%-100%, depending on which two groups were compared. When the different clinical subgroups were separated, MDA parameters were always better than conventional MTR histogram parameters, with P values ranging from.05 to less than 1 x 10(-6) of those attained with the best conventional parameter. CONCLUSION: With MDA, MTR histograms of brain tissue may provide diagnostic information for individual patients in the clinical context of SLE. PMID- 11867792 TI - Multiple sclerosis: diffusion tensor MR imaging for evaluation of normal appearing white matter. AB - PURPOSE: To determine whether the normal-appearing white matter (NAWM) regions surrounding and remote from multiple sclerosis (MS) plaques have abnormal diffusional anisotropy and to compare anisotropy maps with apparent diffusion coefficient (ADC) maps for sensitivity in the detection of white matter (WM) abnormalities. MATERIALS AND METHODS: Conventional and diffusion tensor magnetic resonance (MR) imaging examinations were performed in 26 patients with MS and in 26 age-matched control subjects. Fractional anisotropy (FA) and ADC maps were generated and coregistered with T2-weighted MR images. Uniform regions of interest were placed on plaques, periplaque white matter (PWM) regions, NAWM regions in the contralateral side of the brain, and WM regions in control subjects to obtain FA and ADC values, which were compared across the WM regions. RESULTS: The mean FA was 0.280 for plaques, 0.383 for PWM, 0.493 for NAWM, and 0.537 for control subject WM. The mean ADC was 1.025 x 10(-3) mm(2)/sec for plaques, 0.786 x 10(-3) mm(2)/sec for PWM, 0.739 x 10(-3) mm(2)/sec for NAWM, and 0.726 x 10(-3) mm(2)/sec for control subject WM. Significant differences in anisotropy and ADC values were observed among all WM regions (P <.001 for all comparisons, except ADC in NAWM vs control subject WM [P =.018]). CONCLUSION: The anisotropy and ADC values were abnormal in all WM regions in the patients with MS and were worse in the periplaque regions than in the distant regions. Diffusion tensor MR imaging may be more accurate than T2-weighted MR imaging for assessment of disease burden. PMID- 11867793 TI - Unilateral transpedicular percutaneous vertebroplasty: initial experience. AB - PURPOSE: To compare the radiographic and clinical outcomes of a unipediculate approach with those of standard bipediculate vertebroplasty. MATERIALS AND METHODS: Retrospective review of percutaneous vertebroplasties yielded 18 vertebrae in 17 patients that were treated with a standard bipediculate approach and 57 vertebrae in 32 patients that were treated with a modified unipediculate approach. Anteroposterior radiographs obtained after the procedure were viewed to calculate the percentage of cement opacification in both vertebral halves, and mean values were compared between uni- and bipediculate approaches by using a two tailed Student t test. Clinical outcomes, including pain relief and change in pain medication requirements, were compared in the two groups by using the chi(2) or Fisher exact test. RESULTS: With the unipediculate approach, filling across the midline was achieved in 55 (96%) of 57 injections. Mean opacification of vertebral body halves was 83% +/- 19 (SD) and 77% +/- 16 for the bipediculate and unipediculate approaches, respectively (P =.19). Among patients with available follow-up data, 16 (94%) of 17 patients who underwent the bipediculate procedure and 28 (88%) of 32 patients who underwent the unipediculate procedure achieved adequate pain relief (P =.65) with mean decreases in pain severity of 7.3 +/- 3.1 and 6.6 +/- 2.9, respectively. CONCLUSION: Use of a unipediculate approach in percutaneous vertebroplasty allows filling of both vertebral halves from a single puncture site with no statistically significant difference in clinical outcome from that of bipediculate vertebroplasty. PMID- 11867795 TI - Case 44: Adenocarcinoma of the urachus. PMID- 11867797 TI - Finger pulley injuries in extreme rock climbers: depiction with dynamic US. AB - PURPOSE: To determine the ability of dynamic ultrasonography (US) to depict finger pulley injuries in extreme rock climbers. MATERIALS AND METHODS: Sixty four extreme rock climbers (climbing levels 8-11 on a scale ranging from 1 to 11; Union Internationale des Associations d'Alpinisme) with finger injuries (75 symptomatic and 181 asymptomatic fingers) were examined by using US, with the transducer operating at 12 MHz. The distance between the flexor tendon and phalanx was evaluated in extension and forced flexion at the level of the A2 and A4 annular pulleys as an indicator of tendon bowstringing. A distance between the flexor tendon and phalanx greater than 1.0 mm was interpreted as positive for a pulley injury. US findings were compared with those of magnetic resonance imaging. Surgical correlation was available in seven cases. Statistical analysis was performed by using analysis of variance, the Student t test, and the Bonferroni method. RESULTS: US depicted 16 (100%) of 16 complete A2 pulley ruptures, nine (100%) of nine complete A4 pulley ruptures, six (86%) of seven surgically proved complete combined A2 and A3 pulley ruptures, and 15 (100%) of 15 incomplete A2 pulley ruptures. Measurement of distance between the flexor tendon and phalanx was significantly different among patient subsets without pulley ruptures and those with incomplete, complete, or complete combined pulley ruptures (P <.001). The sensitivity of US for depiction of finger pulley injuries was 98%, and specificity was 100%. CONCLUSION: Dynamic US allows excellent depiction of finger pulley injuries in extreme rock climbers. PMID- 11867796 TI - Blunt abdominal trauma in children: evaluation with emergency US. AB - PURPOSE: To assess the accuracy of emergency abdominal ultrasonography (US) in the detection of both hemoperitoneum and parenchymal organ injury in children. MATERIALS AND METHODS: Imaging findings were recorded prospectively in 744 consecutive children who underwent emergency US from January 1995 to October 1998; free fluid and parenchymal abnormalities of specific organs were also noted. Patients with intraabdominal injuries were identified retrospectively. Computed tomographic (CT) findings, intraoperative findings, and clinical outcome were compared with the initial US findings. Sensitivity, specificity, and positive and negative predictive values were calculated for patients who underwent CT, laparotomy, or both after US. RESULTS: Seventy-five (10%) of 744 patients had intraabdominal injuries, and US depicted free fluid in 42 of them. US had 56% sensitivity, 97% specificity, 82% positive predictive value, and 91% negative predictive value for detection of hemoperitoneum alone. US helped identify parenchymal abnormalities that corresponded to actual organ injury without accompanying free fluid in nine patients (12%). Inclusion of identification of parenchymal organ injury at US increased the sensitivity of US to 68%, with an accuracy of 92%. CONCLUSION: US for blunt abdominal trauma in children is highly accurate and specific, but moderately sensitive, for detection of intraabdominal injury. PMID- 11867799 TI - The tree-in-bud sign. PMID- 11867800 TI - Screening for early lung cancer with low-dose spiral CT: prevalence in 817 asymptomatic smokers. AB - PURPOSE: To present prevalence screening data from a nonrandomized screening trial by using low-dose computed tomography (CT) and a simple algorithm based on the size and attenuation of detected nodules to guide diagnostic work-up. MATERIALS AND METHODS: Eight hundred seventeen asymptomatic volunteers (age range, 40-78 years; median age, 53 years; median tobacco consumption, 45 pack years) underwent spiral low-dose CT of the chest without contrast material enhancement. We regarded all noncalcified pulmonary nodules greater than 10 mm in diameter as potentially malignant and recommended histologic examination or follow-up after 3, 6, 12, and 24 months to exclude growth. For noncalcified pulmonary nodules of 10 mm or smaller, repeat low-dose CT was recommended to exclude growth. RESULTS: In 43% (350 of 817) of individuals, 858 noncalcified pulmonary nodules were found. Thirty-two nodules in 29 subjects were larger than 10 mm. Biopsy of 15 lesions revealed lung cancer in 12 lesions in 11 subjects (prevalence for all ages, 1.3% [11 of 817 subjects]; >50 years of age, 2.1% [11 of 519 subjects]; >60 years of age, 3.9% [eight of 206 subjects]), with a high proportion of early tumor stages (seven tumors, stage I; two, stage II; and three, stage III); three lesions were benign. In 17 nodules larger than 10 mm, follow-up with low-dose CT for a minimum of 24 months did not demonstrate growth. CONCLUSION: Lung cancer screening with low-dose CT demonstrated a prevalence of asymptomatic cancers in 1.3% of a smoking population, including a high proportion of early tumor stages and a 20% (three of 15) rate of invasive procedures for benign lesions. PMID- 11867798 TI - Pisotriquetral joint: assessment with MR imaging and MR arthrography. AB - PURPOSE: To evaluate magnetic resonance (MR) imaging and MR arthrographic findings in the pisotriquetral joint (PTJ) and their contribution to assessment of PTJ osteoarthritis. MATERIALS AND METHODS: Images of 22 fresh human cadaveric PTJs were obtained with both conventional and arthrographic MR techniques. The MR appearances of all intraarticular and periarticular structures were analyzed and correlated with anatomic slices. Two readers graded visibility of anatomic structures and severity of joint abnormalities. Differences in the visibility ratings at standard MR imaging and at MR arthrography were calculated. Association between the type of pisiform insertion of ligament or muscle with cartilaginous abnormalities of the PTJ was assessed. The association between cartilaginous lesions and osteoarthritic changes was calculated. RESULTS: The tendon sheath, the fibrous capsule, and cartilaginous surfaces were better visualized at MR arthrography than at MR imaging. Pisohamate and pisometacarpal ligaments were slightly better seen on MR arthrograms. Tendons, muscles, and retinacular structures were well demonstrated at both conventional MR and MR arthrography. Cartilaginous lesions and osteophytes were easily identified and were detected more often in the pisiform bone than in the triquetral bone. Communication of the PTJ with the radiocarpal joint was noted in 18 (82%) of 22 wrists. CONCLUSION: MR imaging and/or MR arthrography allows visualization of all anatomic structures of the PTJ. MR arthrography improves visualization of findings of osteoarthritis. PMID- 11867801 TI - Pulmonary arterial hypertension: thin-section CT predictors of epoprostenol therapy failure. AB - PURPOSE: To correlate pretherapeutic thin-section computed tomographic (CT) findings in patients with pulmonary hypertension with the risk of fatality with treatment with epoprostenol. MATERIALS AND METHODS: Seventy-three consecutive patients with severe pulmonary hypertension treated with epoprostenol were retrospectively separated into two groups. The first group included 12 patients who had a fatal outcome with epoprostenol therapy. The second group (n = 61) was a reference group of patients with epoprostenol-induced clinical improvement. Pretherapeutic thin-section CT scans of each patient were reviewed. RESULTS: Poorly defined nodular opacities (P =.003), septal lines (P =.04), pleural effusion (P =.01), and adenopathy (P =.009) strongly correlated with a risk of clinical worsening with treatment. In six patients in group 1, postmortem examination of the lung revealed either pulmonary veno-occlusive disease or pulmonary capillary hemangiomatosis. CONCLUSION: On pretherapeutic thin-section CT scans, poorly defined nodular opacities, septal lines, pleural effusion, and adenopathy should raise suspicion for pulmonary veno-occlusive disease or pulmonary capillary hemangiomatosis and provoke possible further evaluation before epoprostenol therapy. PMID- 11867802 TI - Morphologic phenotyping with MR microscopy: the visible mouse. AB - A method for rapid morphologic phenotyping is demonstrated by using magnetic resonance microscopy. Whole fixed C57BL/6J mice were imaged at 110-microm isotropic resolution; limited volumes of the intact specimen, at 50-microm isotropic resolution; and isolated organs, at 25-microm isotropic resolution. The three-dimensional imaging technique was applied to uricase knockout mice to demonstrate the method for the evaluation of morphologic phenotype. PMID- 11867803 TI - Clip migration after 11-gauge vacuum-assisted stereotactic biopsy: case report. AB - A 68-year-old woman underwent stereotactic biopsy of a small cluster of calcifications. The post-biopsy mammograms showed the biopsy-marking clip to be located correctly at the biopsy site. Follow-up mammograms 1 year later showed that the clip migrated to another quadrant of the breast. Findings in this case demonstrate that at long-term follow-up a biopsy-marking clip may not be accurately marking the biopsy site. PMID- 11867804 TI - Percutaneous tumor ablation: increased necrosis with combined radio-frequency ablation and intravenous liposomal doxorubicin in a rat breast tumor model. AB - PURPOSE: To determine whether a combination of intravenous liposomal doxorubicin and radio-frequency (RF) ablation increases tumor destruction compared with RF alone in an animal tumor model. MATERIALS AND METHODS: R3230 mammary adenocarcinoma 1.4-1.8-cm- diameter nodules were implanted subcutaneously in 132 female Fischer rats. Initially, tumors were treated with (a) conventional, monopolar RF (mean, 250 mA +/- 25 [SD] at 70 degrees C +/- 1 for 5 minutes) ablation alone, (b) RF ablation followed by intravenous administration of 1 mg of liposomal doxorubicin, (c) RF ablation followed by intravenous administration of 1 mg of empty liposomes, (d) RF ablation and direct intratumoral administration of liposomal doxorubicin, or (e) no treatment. Subsequently, the dose (0.06-2.00 mg) of liposomal doxorubicin, the timing of administration (3 days before to 3 days after RF ablation), and the time of pathologic examination (0-72 hours after treatment) were varied. RESULTS: Mean coagulation diameter for treated tumors follows: 6.7 mm +/- 0.6, RF ablation alone; 11.1 mm +/- 1.5, RF ablation and intravenous administration of empty liposomes (P <.05, compared with RF ablation alone); and 8.4 mm +/- 1.1, RF ablation with intratumoral administration of liposomal doxorubicin (P <.05, compared with RF ablation alone). Maximal increased mean coagulation diameter (13.1 mm +/- 1.5) was observed with a combination of liposomal doxorubicin and RF ablation (P <.001, for all comparisons). The increased coagulation for combination therapy developed over 48 hours after therapy. Coagulation diameter did not vary with the doxorubicin concentration range and was not dependent on the timing of administration of liposomal doxorubicin from 3 days before to 24 hours after RF ablation. CONCLUSION: Intravenous administration of liposomal doxorubicin can improve RF ablation, since it increases coagulation diameter in solid tumors compared with RF ablation alone or a combination of RF ablation with administration of empty liposomes. PMID- 11867805 TI - Radio-frequency tumor ablation: internally cooled electrode versus saline enhanced technique in an aggressive rabbit tumor model. AB - PURPOSE: To compare two methods of radio-frequency (RF) ablation, saline enhancement technique and internally cooled electrodes, for the treatment of small breast cancers in an animal model--highly aggressive VX2 rabbit tumors surrounded by adipose tissue. MATERIALS AND METHODS: Twenty-seven tumors were implanted into retroperitoneal fat of 14 New Zealand White rabbits. RF ablation was performed with ultrasonographic (US) guidance after tumors had grown to 15 mm. Fourteen tumors in seven animals were treated with internally cooled electrodes (30-mm-tip single electrode, 60 W, 10 min); 13 tumors in seven animals, with saline enhancement (0.5 mL/min of saline, 25-mm tip, 30 W, 10 min). Autopsy and histopathologic assessment were performed 3 weeks after therapy. RESULTS: Real-time US of RF ablation was not possible with either method because of obscuration by the increasing hyperechogenicity of the tumor and the surrounding adipose tissue. Equivalent efficacy was demonstrated with the two methods. Significantly greater complications were observed with the saline technique: Free retroperitoneal fluid was detected in one of seven animals with internally cooled electrodes and in all seven animals with saline enhancement (P <.01). Damage to remote structures such as the kidney, spine muscle, and skin was observed at autopsy in one of seven animals with internally cooled technique versus five of seven with saline enhancement (P <.01). CONCLUSION: Given a lower complication rate and similar treatment efficacy in an animal tumor model, internally cooled RF electrode may be advantageous to adjuvant saline infusion for the minimally invasive treatment of breast tumors. PMID- 11867806 TI - Imaging of differential protease expression in breast cancers for detection of aggressive tumor phenotypes. AB - PURPOSE: To determine if different expression levels of tumor cathepsin-B activity in well differentiated and undifferentiated breast cancers could be revealed in vivo with optical imaging. MATERIALS AND METHODS: A well differentiated human breast cancer (BT20, n = 8) and a highly invasive metastatic human breast cancer (DU4475, n = 8) were implanted orthotopically in athymic nude mice. Tumor-bearing animals were examined in vivo with near-infrared fluorescence (NIRF) imaging 24 hours after intravenous injection of an enzyme-sensing imaging probe. Immunohistochemistry, Western blotting (on cells and whole tumor samples), and correlative fluorescence microscopy were performed. RESULTS: Both types of breast cancers activated the NIRF probe so that tumors became readily detectable. However, in tumors of equal size, there was a 1.5-fold higher fluorescence signal in the highly invasive breast cancer (861 arbitrary units +/- 88) compared with the well differentiated lesion (566 arbitrary units +/- 36, P <.01). Western blotting confirmed a higher cathepsin-B protein content in the highly invasive breast cancer (DU4475) of about 1.4-fold (whole tumor samples) to 1.7-fold (cells). Immunohistochemistry and fluorescence microscopy findings confirmed the imaging findings. CONCLUSION: Cathepsin-B enzyme activity can be determined in vivo with NIRF optical imaging, while differences in tumoral expression may correlate with tumor aggressiveness. PMID- 11867807 TI - Minimizing diaphragmatic injury during radio-frequency ablation: efficacy of subphrenic peritoneal saline injection in a porcine model. AB - PURPOSE: To investigate if a targeted subphrenic peritoneal infusion of normal saline to separate liver from diaphragm before radio-frequency (RF) ablation could minimize or eliminate diaphragmatic injury. MATERIALS AND METHODS: With a 2 cm-diameter, eight-prong RF needle electrode, 37 hepatic dome RF lesions were created in 10 pigs. Seventeen lesions were created before (non-saline group) and 20 lesions after (post-saline group) intraperitoneal infusion of approximately 500 mL of normal saline. Ten non-saline lesions were created deep (centered 1-2 cm from the liver surface) and seven superficially (centered within 1 cm of the capsule). All 20 post-saline lesions were created superficially. Helical enhanced computed tomography was performed 24-48 hours after ablation. The pigs were killed immediately, and the diaphragm was visually inspected and sectioned. Diaphragmatic injury was graded as 0, no injury; 1, injury up to one-third thickness; 2, injury to two-thirds thickness; 3, full-thickness injury. Representative grade 3 injuries and all partial injuries underwent gross and histologic analysis. RESULTS: All 10 deep non-saline RF lesions caused grade 0 injury. All seven superficial non-saline lesions caused grade 3 injury. Of the 20 superficial post-saline lesions, 13 (65%) caused grade 0 injury; four (20%), grade 1; and three (15%), grade 3. The post-saline group caused significantly less diaphragmatic injury (P <.05). CONCLUSION: Intraperitoneal saline infusion may reduce the frequency and severity of diaphragmatic injury when adjacent liver is treated with RF ablation. PMID- 11867808 TI - Hepatic tumor detection: MR imaging and conventional US versus pulse-inversion harmonic US of NC100100 during its reticuloendothelial system-specific phase. AB - PURPOSE: To compare conventional ultrasonography (US) and magnetic resonance (MR) imaging with contrast agent-enhanced US for detection of VX-2 liver tumors in rabbits. MATERIALS AND METHODS: Conventional gray-scale liver US was performed in 65 rabbits, 38 of which had VX-2 hepatic tumor implants. Twenty minutes after contrast agent injection, gray-scale pulse-inversion harmonic US images of the liver-specific phase were obtained. Following sacrifice of the animals, T1- and T2-weighted MR imaging was performed at 4-mm intervals. Pathologic analysis was performed as the reference standard. The capability of each imaging modality to correctly depict tumor presence or absence and the number of tumors was compared. RESULTS: Conventional US correctly depicted the presence or absence of tumors in 54 rabbits, for an accuracy of 83%, sensitivity of 71%, and specificity of 100%. With contrast-enhanced US, accuracy increased to 92% (60 correct cases); sensitivity, to 87%; and specificity, to 100%. MR imaging facilitated 56 correct diagnoses, for an accuracy of 86%, sensitivity of 82%, and specificity of 93%. There was a marginally significant difference between US with and US without contrast agent (P =.07) but not between MR imaging and contrast-enhanced US (P > or = .34). When the numbers of correctly detected tumors were compared, contrast enhanced US performed significantly better than MR imaging (P =.02) and conventional US (P =.04). CONCLUSION: There was no significant difference between contrast-enhanced US and MR imaging in the detection of hepatic tumors, whereas contrast-enhanced US had the highest accuracy (92%) of the three modalities studied. PMID- 11867809 TI - Comparison of MR imaging breast coils. AB - In one volunteer, five breast coils were evaluated for signal-to-noise ratio (SNR), uniformity, comfort, subject orientation, access to the breast, and unilateral imaging options. The four-coil arrays provided superior SNR, imaging flexibility, and access. Uniformity and comfort were issues with all coils. Substantial design differences exist between coils; purchasers should ensure that their specific requirements are met. PMID- 11867811 TI - Spinal dural arteriovenous fistula localization with a technique of first-pass gadolinium-enhanced MR angiography: initial experience. AB - Nine patients with initial magnetic resonance (MR) imaging and clinical findings suggestive of spinal dural arteriovenous fistula (AVF) underwent spinal MR angiography with an autotriggered elliptic centric ordered three-dimensional gadolinium-enhanced technique (hereafter, this MR angiographic technique) before conventional intraarterial angiography. In all nine patients, findings with this MR angiographic technique correctly and precisely localized the spinal dural AVF. Observer error resulted in one case in which the site of the fistula was not prospectively reported but was easily identified retrospectively on the spinal MR angiogram. PMID- 11867810 TI - Cardiac function: MR evaluation in one breath hold with real-time true fast imaging with steady-state precession. AB - In 12 healthy volunteers and eight patients with cardiac disease, cine magnetic resonance (MR) imaging in the heart was performed with real-time true fast imaging with steady-state precession (FISP), which permitted evaluation of the entire left ventricle in one breath hold (91 msec per frame, 13 frames per section position, nine short-axis section positions per breath hold). Contrast-to noise ratios (CNRs) and left ventricular mass and function measurements with this technique were compared in all subjects with single-section true FISP imaging and, in the volunteers only, with segmented fast low-angle shot (FLASH) MR imaging. Myocardium-to-blood CNR was significantly higher for both true FISP sequences compared with the FLASH sequence. Measurements of resting left ventricular function with real-time true FISP imaging were comparable with those derived from a series of separate breath-hold single-section true FISP acquisitions. PMID- 11867812 TI - Vertebroplasty: reusable flange converter with hub lock for injection of polymethylmethacrylate with screw-plunger syringe. AB - During percutaneous vertebroplasty, a screw-plunger syringe provides a powerful controlled injection, but coupling of the screw plunger with interchangeable disposable standard syringe barrels results in syringe flange bending failure during the middle of the injection. A flange converter produced from stainless steel and a hub lock produced from plastic or aluminum were used during vertebroplasty at 172 levels in 86 patient-treatment sessions. The flange converter and hub lock increase the achievable volume that can be injected, can be fabricated with hand machining, and cost much less than other commercially available devices. PMID- 11867813 TI - Importance of quality breast imaging in symptomatic women. PMID- 11867814 TI - Negative mammographic and US findings do not help exclude breast cancer. PMID- 11867815 TI - Intraarterial MR angioplasty in iliac arterial stenosis. PMID- 11867819 TI - The dawn of the SPPARMs? AB - Thiazolidinediones (TZDs) are used as antidiabetic agents in the treatment of type II diabetes. These compounds are ligands for the nuclear hormone receptor PPARgamma, which is highly expressed in adipose tissue. PPARgamma acts as a molecular switch in the process of fat cell development. The quest for the ideal antidiabetic agent is challenged by the need to develop PPARgamma ligands that improve insulin sensitivity, but do not promote fat cell formation. A newly described PPARgamma ligand may represent an initial step in this direction and could lead to improved agents for treating insulin resistance in type II diabetes. PMID- 11867820 TI - Macrolide antibiotics and cystic fibrosis. PMID- 11867821 TI - Psychological factors in asthma control and attack risk. PMID- 11867822 TI - Non-invasive ventilation in acute respiratory failure. PMID- 11867823 TI - Effect of long term treatment with azithromycin on disease parameters in cystic fibrosis: a randomised trial. AB - BACKGROUND: Relentless chronic pulmonary inflammation is the major contributor to morbidity and mortality in patients with cystic fibrosis (CF). While immunomodulating therapies such as prednisolone and ibuprofen may be beneficial, their use is limited by side effects. Macrolides have immunomodulatory properties and long term use dramatically improves prognosis in diffuse panbronchiolitis, a condition with features in common with the lung disease of CF. METHODS: To determine if azithromycin (AZM) improves clinical parameters and reduces inflammation in patients with CF, a 3 month prospective randomised double blind, placebo controlled study of AZM (250 mg/day) was undertaken in adults with CF. Monthly assessment included lung function, weight, and quality of life (QOL). Blood and sputum collection assessed systemic inflammation and changes in bacterial flora. Respiratory exacerbations were treated according to the policy of the CF Unit. RESULTS: Sixty patients were recruited (29 men) of mean (SD) age 27.9 (6.5) years and initial forced expiratory volume in 1 second (FEV1) 56.6 (22.3)% predicted. FEV1% and forced vital capacity (FVC)% predicted were maintained in the AZM group while in the placebo group there was a mean (SE) decline of -3.62 (1.78)% (p=0.047) and -5.73 (1.66)% (p=0.001), respectively. Fewer courses of intravenous antibiotics were used in patients on AZM (0.37 v 1.13, p=0.016). Median C reactive protein (CRP) levels declined in the AZM group from 10 to 5.4 mg/ml but remained constant in the placebo group (p<0.001). QOL improved over time in patients on AZM and remained unchanged in those on placebo (p=0.035). CONCLUSION: AZM in adults with CF significantly improved QOL, reduced CRP levels and the number of respiratory exacerbations, and reduced the rate of decline in lung function. Long term AZM may have a significant impact on morbidity and mortality in patients with CF. Further studies are required to define frequency of dosing and duration of benefit. PMID- 11867824 TI - Attack context: an important mediator of the relationship between psychological status and asthma outcomes. AB - BACKGROUND: The importance of psychosocial variables in asthma is increasingly recognised, although attempts to relate these to asthma outcomes often produce only weak relationships. This study aimed to identify whether such relationships might be obscured by the effects of recent asthma experience on psychological status. METHODS: An adult community sample of 37 patients who had suffered a recent attack of asthma and 37 with stable asthma were given measures of panic fear and control confidence. The relationship with subsequent emergency service use was examined using two way ANOVA and correlational analyses. Covariate influences (psychiatric morbidity, age, sex, treatment level, asthma duration, social status) were considered. RESULTS: Control confidence predicted emergency service use in different ways for recent attack and stable asthma patients. This interaction was highly significant (F(1,69) = 10.32, p<0.005) with high confidence relating to an increased risk of an attack in the recent attack group and low confidence relating to increased risk for the stable asthma group. There was also an interaction between panic fear and attack context (F(1,69) = 11.05, p<0.005) with low panic fear resulting in more attacks for recent attack cases. CONCLUSIONS: Attack context (having a recent attack) is an important mediator of psychological status. Strong cognitive/affective responses to attacks may motivate improved self-care and this represents a window of opportunity for self care interventions. Weak cognitive/affective responses to attacks may reflect denial and require different intervention approaches. For those with recently stable asthma the relationships are qualitatively and quantitatively different, and the implications for intervention are also discussed. PMID- 11867825 TI - Laser acupuncture in children and adolescents with exercise induced asthma. AB - BACKGROUND: Laser acupuncture, a painless technique, is a widely used alternative treatment method for childhood asthma, although its efficacy has not been proved in controlled clinical studies. METHODS: A double blind, placebo controlled, crossover study was performed to investigate the possible protective effect of a single laser acupuncture treatment on cold dry air hyperventilation induced bronchoconstriction in 44 children and adolescents of mean age 11.9 years (range 7.5-16.7) with exercise induced asthma. Laser acupuncture was performed on real and placebo points in random order on two consecutive days. Lung function was measured before laser acupuncture, immediately after laser acupuncture (just before cold dry air challenge (CACh)), and 3 and 15 minutes after CACh. CACh consisted of a 4 minute isocapnic hyperventilation of -10 degrees C absolute dry air. RESULTS: Comparison of real acupuncture with placebo acupuncture showed no significant differences in the mean maximum CACh induced decrease in forced expiratory volume in 1 second (27.2 (18.2)% v 23.8 (16.2)%) and maximal expiratory flow at 25% remaining vital capacity (51.6 (20.8)% v 44.4 (22.3)%). CONCLUSIONS: A single laser acupuncture treatment offers no protection against exercise induced bronchoconstriction in paediatric and adolescent patients. PMID- 11867826 TI - Influence of cigarette smoking on inhaled corticosteroid treatment in mild asthma. AB - BACKGROUND: Although inhaled corticosteroids have an established role in the treatment of asthma, studies have tended to concentrate on non-smokers and little is known about the possible effect of cigarette smoking on the efficacy of treatment with inhaled steroids in asthma. A study was undertaken to investigate the effect of active cigarette smoking on responses to treatment with inhaled corticosteroids in patients with mild asthma. METHODS: The effect of treatment with inhaled fluticasone propionate (1000 microg daily) or placebo for 3 weeks was studied in a double blind, prospective, randomised, placebo controlled study of 38 steroid naive adult asthmatic patients (21 non-smokers). Efficacy was assessed using morning and evening peak expiratory flow (PEF) readings, spirometric parameters, bronchial hyperreactivity, and sputum eosinophil counts. Comparison was made between responses to treatment in non-smoking and smoking asthmatic patients. RESULTS: There was a significantly greater increase in mean morning PEF in non-smokers than in smokers following inhaled fluticasone (27 l/min v -5 l/min). Non-smokers had a statistically significant increase in mean morning PEF (27 l/min), mean forced expiratory volume in 1 second (0.17 l), and geometric mean PC20 (2.6 doubling doses), and a significant decrease in the proportion of sputum eosinophils (-1.75%) after fluticasone compared with placebo. No significant changes were observed in the smoking asthmatic patients for any of these parameters. CONCLUSIONS: Active cigarette smoking impairs the efficacy of short term inhaled corticosteroid treatment in mild asthma. This finding has important implications for the management of patients with mild asthma who smoke. PMID- 11867827 TI - Occupational asthma due to low molecular weight agents: eosinophilic and non eosinophilic variants. AB - BACKGROUND: Despite having a work related deterioration in peak expiratory flow (PEF), many workers with occupational asthma show a low degree of within day diurnal variability atypical of non-occupational asthma. It was hypothesised that these workers would have a neutrophilic rather than an eosinophilic airway inflammatory response. METHODS: Thirty eight consecutive workers with occupational asthma induced by low molecular weight agents underwent sputum induction and assessment of airway physiology while still exposed at work. RESULTS: Only 14 (36.8%) of the 38 workers had sputum eosinophilia (>2.2%). Both eosinophilic and non-eosinophilic groups had sputum neutrophilia (mean (SD) 59.5 (19.6)% and 55.1 (18.8)%, respectively). The diurnal variation and magnitude of fall in PEF during work periods was not significantly different between workers with and without sputum eosinophilia. Those with eosinophilia had a lower forced expiratory volume in 1 second (FEV1; 61.4% v 83% predicted, mean difference 21.6, 95% confidence interval (CI) 9.2 to 34.1, p=0.001) and greater methacholine reactivity (geometric mean PD20 253 microg v 1401 microg, p=0.007). They also had greater bronchodilator reversibility (397 ml v 161 ml, mean difference 236, 95% CI of difference 84 to 389, p=0.003) which was unrelated to differences in baseline FEV(1). The presence of sputum eosinophilia did not relate to the causative agent, duration of exposure, atopy, or lack of treatment. CONCLUSIONS: Asthma caused by low molecular weight agents can be separated into eosinophilic and non-eosinophilic pathophysiological variants with the latter predominating. Both groups had evidence of sputum neutrophilia. Sputum eosinophilia was associated with more severe disease and greater bronchodilator reversibility but no difference in PEF response to work exposure. PMID- 11867829 TI - Correlation between the bronchial subepithelial layer and whole airway wall thickness in patients with asthma. AB - BACKGROUND: The epithelial reticular basement membrane (Rbm) of the airway wall thickens in patients with asthma. However, whether the thickening parallels whole airway wall thickening, which limits airflow, is unknown. The aim of this study was to examine the correlation between the bronchial Rbm thickening and whole airway wall thickening in asthma. In addition, the association of Rbm and whole wall thickening with airflow obstruction was examined. METHODS: Forty nine patients with asthma and 18 healthy control subjects took part in the study. The Rbm thickness was measured in bronchial biopsy specimens and whole airway wall thickness was assessed with high resolution computed tomographic (HRCT) scanning after pretreatment with oral steroids for 2 weeks and inhaled beta2 agonist to minimise reversible changes of the airway walls. The percentage airway wall area (WA%; defined as (wall area/total airway area) x 100) and percentage airway wall thickness (WT%; defined as [(ideal outer diameter - ideal luminal diameter)/ideal outer diameter] x 100) were determined from HRCT scans to assess whole airway wall thickness. Spirometric tests were also performed. RESULTS: WA% and WT% were higher in patients with asthma than in healthy subjects. Both WA% and WT% were strongly correlated with Rbm thickness. Moreover, these three indices of airway wall thickness were inversely correlated with the percentage of predicted forced expiratory volume in 1 second in patients with asthma. CONCLUSIONS: These findings indicate that Rbm thickening parallels whole airway wall thickening which can cause irreversible airflow obstruction in patients with asthma. PMID- 11867828 TI - Additive anti-inflammatory effect of formoterol and budesonide on human lung fibroblasts. AB - BACKGROUND: It has been shown that treatment with a long acting beta2 agonist in addition to a glucocorticoid is beneficial in the treatment of asthma. In asthma inflammatory cells, particularly eosinophils, migrate into the pulmonary tissue and airway lumen by means of adhesion molecules expressed on resident tissue cells--that is, fibroblasts--and become activated by cytokines and adhesive interactions. A study was undertaken to determine whether an interaction exists between the long acting beta2 agonist formoterol and the glucocorticoid budesonide on inhibition of adhesion molecule expression, as well as chemo/cytokine production by human lung fibroblasts. METHODS: Lung fibroblasts were preincubated with therapeutically relevant drug concentrations of 10(-8) M to 10(-10) M. Cells were stimulated with interleukin (IL)-1beta (1 or 10 U/ml) for 8 hours and supernatants were collected for measurement of GM-CSF and IL-8 concentrations. The cells were fixed and subjected to a cell surface ELISA technique to measure the expression of ICAM-1 and VCAM-1. RESULTS: Formoterol exerted an additive effect on the inhibition of IL-1beta stimulated ICAM-1 and VCAM-1 upregulation and GM-CSF production by budesonide in concentrations of 10( 9) M and above (p<0.05). IL-8 production was not influenced by formoterol. CONCLUSION: Formoterol exerts an additive effect on the anti-inflammatory properties of budesonide. In vitro data support the finding that the combination of budesonide and formoterol in asthma treatment strengthens the beneficial effect of either drug alone. PMID- 11867830 TI - High resolution computed tomographic assessment of airway wall thickness in chronic asthma: reproducibility and relationship with lung function and severity. AB - BACKGROUND: In some patients chronic asthma results in irreversible airflow obstruction. High resolution computed tomography (HRCT) has been advocated for assessing the structural changes in the asthmatic lung and permits investigation of the relationships between airway wall thickening and clinical parameters in this condition. METHODS: High resolution CT scanning was performed in 49 optimally controlled asthmatic patients and measurements of total airway and lumen diameter were made by two independent radiologists using electronic callipers. Wall area as % total airway cross sectional area (WA%) and wall thickness to airway diameter ratio (T/D) were calculated for all airways clearly visualised with a transverse diameter of more than 1.5 mm, with a mean value derived for each patient. Intra- and inter-observer variability was assessed for scope of agreement in a subgroup of patients. Measurements were related to optimum forced expiratory volume in 1 second (FEV1), forced mid expiratory flow, carbon monoxide gas transfer, two scores of asthma severity, airway inflammation as assessed with induced sputum, and exhaled nitric oxide. RESULTS: Neither observer produced a statistically significant difference between measurements performed on two occasions but there was a significant difference between observers (limits of agreement -2.6 to 6.8 for WA%, p<0.0001). However, mean WA% measured on two occasions differed by no more than 5.4% (limits of agreement -4.0 to 5.4; mean (SD) 0.7 (2.4)). Statistically significant positive associations were observed between both WA% and T/D ratio and asthma severity (r(S)=0.29 and 0.30, respectively, for ATS score), and an inverse association with gas transfer coefficient was observed (r(S)=-0.43 for WA% and r(S)=-0.41 for T/D). No association was identified with FEV1 or airway inflammation. CONCLUSIONS: The airway wall is thickened in more severe asthma and is associated with gas transfer coefficient. This thickening does not relate directly to irreversible airflow obstruction as measured with FEV1. PMID- 11867831 TI - Inspiratory muscle maximum relaxation rate measured from submaximal sniff nasal pressure in patients with severe COPD. AB - BACKGROUND: Slowing of the inspiratory muscle maximum relaxation rate (MRR) is a useful index of severe inspiratory muscle loading and potential fatigue and has been measured from the oesophageal pressure during sniffs in patients with chronic obstructive pulmonary disease (COPD). The purpose of this study was to investigate whether it is possible to measure MRR and detect slowing using sniff nasal pressure in patients with COPD and to investigate the relationship between sniff oesophageal and sniff nasal MRR. METHODS: Eight patients with severe COPD (mean FEV1 0.7 l; 26% predicted) were studied. Each subject performed submaximal sniff manoeuvres before and after walking to a state of severe dyspnoea on a treadmill. Oesophageal and gastric pressures were measured using balloon tipped catheters and nasal pressure was measured using an individually modelled nasal cast. MRR (% pressure fall/10 ms) was determined for each sniff and any change following exercise was reported as percentage of baseline to allow comparison of sniff nasal and oesophageal MRR. RESULTS: At rest the mean (SE) sniff Poes MRR was 7.1 (0.3) and the mean Pnasal MRR was 8.6 (0.1). At 1 minute following exercise there was a mean decrease in sniff Poes MRR of 33.7% (range 20.7-53.4%) and a mean decrease in sniff Pnasal MRR of 28.2% (range 8.1-52.8%). The degree of slowing and time course of recovery was similar, with both returning to baseline values within 5-10 minutes. A separate analysis of the sniff pressures using only the nasal pressure traces demonstrated a similar pattern of slowing and recovery. CONCLUSIONS: It is possible to detect slowing of the inspiratory muscles non invasively using sniff nasal pressures in patients with COPD. This could be a useful technique with which to measure severe and potentially fatiguing inspiratory muscle loading, both in clinical settings and during exercise studies. PMID- 11867832 TI - Effect of assist negative pressure ventilation by microprocessor based iron lung on breathing effort. AB - BACKGROUND: The lack of patient triggering capability during negative pressure ventilation (NPV) may contribute to poor patient synchrony and induction of upper airway collapse. This study was undertaken to evaluate the performance of a microprocessor based iron lung capable of thermistor triggering. METHODS: The effects of NPV with thermistor triggering were studied in four normal subjects and six patients with an acute exacerbation of chronic obstructive pulmonary disease (COPD) by measuring: (1) the time delay (TDtr) between the onset of inspiratory airflow and the start of assisted breathing; (2) the pressure-time product of the diaphragm (PTPdi); and (3) non-triggering inspiratory efforts (NonTrEf). In patients the effects of negative extrathoracic end expiratory pressure (NEEP) added to NPV were also evaluated. RESULTS: With increasing trigger sensitivity the mean (SE) TDtr ranged from 0.29 (0.02) s to 0.21 (0.01) s (mean difference 0.08 s, 95% CI 0.05 to 0.12) in normal subjects and from 0.30 (0.02) s to 0.21 (0.01) s (mean difference 0.09 s, 95% CI 0.06 to 0.12) in patients with COPD; NonTrEf ranged from 8.2 (1.8)% to 1.2 (0.1)% of the total breaths in normal subjects and from 11.8 (2.2)% to 2.5 (0.4)% in patients with COPD. Compared with spontaneous breathing, PTPdi decreased significantly with NPV both in normal subjects and in patients with COPD. NEEP added to NPV resulted in a significant decrease in dynamic intrinsic PEEP, diaphragm effort exerted in the pre-trigger phase, and NonTrEf. CONCLUSIONS: Microprocessor based iron lung capable of thermistor triggering was able to perform assist NPV with acceptable TDtr, significant unloading of the diaphragm, and a low rate of NonTrEf. NEEP added to NPV improved the synchrony between the patient and the ventilator. PMID- 11867833 TI - Serum sialyl Lewis X-i antigen in lung adenocarcinoma and idiopathic pulmonary fibrosis. AB - BACKGROUND: Sialyl Lewis X-i antigen has been used as a diagnostic tool for lung adenocarcinoma. However, serum levels of the antigen are also raised in some patients with idiopathic pulmonary fibrosis (IPF) without coexistent malignancy. Expression of the antigen in serum samples of patients with lung adenocarcinoma was evaluated and compared with that of patients with IPF by Western blotting in order to establish a specific laboratory test to differentiate lung adenocarcinoma from IPF. METHODS: The pattern of antigen expression in serum samples from 23 patients with either lung adenocarcinoma or non-malignant lung disease in whom serum levels of sialyl Lewis X-i antigen (>50 U/ml) were significantly increased was studied by Western blotting. RESULTS: Thirteen of the 14 serum samples from patients with lung adenocarcinoma had a molecular weight band at 120 or 130 kD, while five of the six patients with IPF had two or three bands at <97.4 kD. The pattern of antigen expression was apparently different between the two diseases. The sensitivity, specificity, positive likelihood ratio, and negative likelihood ratio of this test in 20 patients with lung adenocarcinoma and IPF were 92.9%, 83.3%, 5.57, and 0.09, respectively. CONCLUSIONS: Western blotting analysis of serum samples from patients with raised levels of sialyl Lewis X-i antigen provides some clinical information for a differential diagnosis between lung adenocarcinoma and IPF. PMID- 11867834 TI - The pulmonary physician in critical care . 3: critical care management of community acquired pneumonia. AB - Severe community acquired pneumonia carries a high mortality. Early recognition of the severity of the illness, rapid and appropriate resuscitation, targeted antibiotic treatment, and the critical care support of multiple failing organ systems are all important in this group of patients. Only by improving all these aspects of care is it likely that survival will increase. PMID- 11867835 TI - Proportional assist ventilation (PAV): a significant advance or a futile struggle between logic and practice? AB - Proportional assist ventilation is a promising addition to other more conventional modes of mechanical ventilation with the theoretical advantage of improving patient-ventilator interaction. It may also be of use as a diagnostic tool in the control of breathing in mechanically ventilated patients. PMID- 11867837 TI - Mechanical insufflation. PMID- 11867836 TI - BAL findings in a patient with pulmonary alveolar proteinosis successfully treated with GM-CSF. AB - BACKGROUND: Idiopathic pulmonary alveolar proteinosis (PAP) has recently been recognised as a disease of impaired alveolar macrophage function caused by neutralising anti-granulocyte-macrophage colony-stimulating (anti-GM-CSF) autoantibodies. Subcutaneous recombinant human GM-CSF is a novel treatment for PAP, but its mechanism of action is unclear. METHODS: Clinical, functional, and bronchoalveolar lavage (BAL) findings were prospectively evaluated in a patient with PAP treated with daily subcutaneous GM-CSF 8 microg/kg for 12 weeks. RESULTS: Treatment resulted in improvements in dyspnoea, lung function, and peak cycle ergometry performance. In serum and BAL fluid the titre of anti-GM-CSF autoantibodies was raised at baseline and markedly reduced on treatment. At baseline the BAL fluid cellular profile showed a decrease in the absolute number and the percentage of macrophages (50%) and an increase in lymphocytes (45%), predominantly CD4+. This cellular distribution remained unchanged after 6 and 12 weeks of treatment while macrophages became morphologically normal and functionally improved. Extracellular proteinaceous material completely disappeared. CONCLUSIONS: Clinically successful treatment of PAP with GM-CSF was associated with a profound reduction in GM-CSF neutralising autoantibodies, improvement in alveolar macrophage morphology and function, but persistent BAL lymphocytosis. PMID- 11867838 TI - Diagnosing TB. PMID- 11867839 TI - Omega-3s and childhood asthma. PMID- 11867840 TI - Inhaled sodium cromoglycate in children with asthma. PMID- 11867841 TI - Current issues for the prevention and treatment of iron deficiency anemia. PMID- 11867842 TI - Effect of delayed cord clamping on iron stores in infants born to anemic mothers: a randomized controlled trial. AB - OBJECTIVE: To study the effects of cord clamping on iron stores of infants born to anemic mothers at 3 months of age. DESIGN: Randomized controlled trial. SETTING: Teaching hospital. METHODS: Infants born to mothers with hemoglobin (Hb)<100 g/L were randomized at delivery to either immediate cord clamping (early group) or cord clamping delayed till descent of placenta into vagina (delayed group). The outcome measures were infant's hemoglobin and serum ferritin 3 months after delivery. RESULTS: There were 102 neonates randomized to early (n = 43) or delayed cord clamping (n = 59). The groups were comparable for maternal age, parity, weight and supplemental iron intake, infant s birth weight, gestation and sex. The mean infant ferritin and Hb at 3 months were significantly higher in the delayed clamping group (118.4 microg/L and 99 g/L) than in the early clamping group (73 microg/L and 88 g/L). The mean decrease in Hb (g/L) at 3 months adjusted for co-variates was significantly less in the delayed clamping group compared to the early clamping group (-1.09, 95% CI-1.58 to -0.62, p >0.001). The odds for anemia (<100 g/L) at 3 months was 7.7 (95% CI 1.84-34.9) times higher in the early compared to the delayed clamping group. CONCLUSION: Iron stores and Hb in infancy can be improved in neonates born to anemic mothers by delaying cord clamping at birth. PMID- 11867843 TI - Iron status of children aged 9-36 months in an urban slum Integrated Child Development Services project in Delhi. AB - OBJECTIVE: To assess the magnitude/severity and possible etiology of anemia and iron deficiency among children 9-36 months of age. METHODS: A population-based study on the prevalence, etiology of anemia and iron status in 545 children, 9-36 months of age, was conducted in an urban slum ICDS (Integrated Child Development Services) project in North-East Delhi. Hemoglobin and serum ferritin was estimated and information on socio-economic, demographic, parasitic infection/infestation and dietary intake was collected. RESULTS: Prevalence of anemia (using WHO cut-off values of Hb >11.0 g/dl) among children, 9-36 months of age, was 64%, of these 7.8% had severe anemia (Hb >7.0 g/dl). Using 10.0 g/dl as the Hb cut-off point 44% children less than 18 months of age in the present study population were anemic. On a sub-sample study, 88% children were estimated to be iron deficient, with serum ferritin concentration less than 12 microg/L. The peripheral smear red cell morphology showed 33.9% as microcytic-hypochromic and 37.1% as dimorphic. Dimorphic anemia was 55% in moderate anemia group. The energy and iron intakes were 56% and 45%, respectively of the Recommended Dietary Allowances (RDA). The parasitic infestation/infection was not related to the prevalence or severity of anemia. CONCLUSION: In Delhi, high prevalence of moderate to severe anemia and iron deficiency with vitamins folate and/or B12 among children under 3 years of age in an ICDS block in operation for 20 years is of concern. Dietary origin was the main cause of anemia in this age group. PMID- 11867844 TI - Management of empyema thoracic in children. PMID- 11867845 TI - Chronic lung disease in neonates: emerging problem in India. PMID- 11867846 TI - Atypical hemolytic uremic syndrome. PMID- 11867847 TI - Lipid profile in children aged 5 to 15 years with parental history of ischemic heart disease, hypertension and diabetes mellitus. PMID- 11867848 TI - Initiating the process of relactation: an Institute based study. PMID- 11867849 TI - Efficacy of nimesulide in pain relief after day care surgery. PMID- 11867851 TI - Fulminant hepatitis A in children with sickle cell disease. PMID- 11867850 TI - Treatment of Menkes disease with parenteral copper histidine. PMID- 11867852 TI - Liver transplantation for fulminant hepatitis A infection. PMID- 11867854 TI - Nephrogenic diabetes insipidus with intracranial calcifications. PMID- 11867856 TI - Scleroderma--CREST syndrome. PMID- 11867853 TI - Neonatal hypernatremia due to high breast-milk sodium. PMID- 11867855 TI - Cutis marmorata telangiectatica congenita. PMID- 11867857 TI - Association of crackles and/or wheezing with tachypnea or chest indrawing in children with pneumonia. PMID- 11867858 TI - Non-bullous ichthyosiform erythroderma with rickets. PMID- 11867859 TI - The dilemma of removing umbilical venous catheters in high-risk neonates with nosocomial sepsis. PMID- 11867860 TI - Early diagnosis of Duchenne muscular dystrophy with high level of transaminases. PMID- 11867861 TI - Measles without rash and encephalopathy: more information is the need of the hour. PMID- 11867862 TI - Other option in bronchiolitis. PMID- 11867863 TI - Intrapartum intervention for prevention of HIV transmission. PMID- 11867865 TI - Autoimmunity and chronic pancreatitis: a concealed relationship. PMID- 11867866 TI - Trypsin activation peptide (TAP) in acute pancreatitis: from pathophysiology to clinical usefulness. PMID- 11867867 TI - Primary culture of porcine pancreatic acinar cells. AB - OBJECTIVE: To develop a method for the primary culture of porcine pancreatic acinar cells. INTERVENTIONS: Dispersed pancreatic acinar cells available utilizing RPMI-1640 medium containing collagenase III. After purification, the isolated acinar cells were cultured in RPMI-1640 medium with the addition of 2.5% fetal bovine serum. MAIN OUTCOME MEASURES: The morphological characteristics of acinar cells were described. (3)H-thymidine incorporation of acinar cells and the activity of amylase or lipase were determined during the culture process. RESULTS: There were no remarkable morphological changes in the pancreatic acinar cells during the 20 days culture. The acini showed a tendency to gather but did not attach to the walls of the culture disks. A good (3)H-thymidine incorporation of acinar cells in the primary culture was maintained. The secretion of amylase or lipase from the acini decreased with the length of time of the culture. DISCUSSION: The primary culture of acinar cells from a porcine pancreas which was carried out in this study maintained the normal morphology of the acinar cells and their ability to grow but not their secretion of amylase or lipase. The method would benefit by the further experiments on acini of porcine pancreas. PMID- 11867869 TI - Respiratory failure. AB - Acute lung injury (ALI) and acute respiratory distress syndrome (ARDS) are common causes of hypoxemic respiratory failure. Multiple etiologies lead to direct and indirect pulmonary injury that progresses through an acute exudative phase, fibroproliferative phase, and recovery phase. Inflammatory mechanisms are thought to play a predominant role in the pathophysiology of ALI/ARDS. Mechanical ventilation with a lower tidal volume and an inspiratory plateau pressure of < or = 30 cm H2O is one intervention that has demonstrated a reduction in mortality. A clinical trial to determine the role of restrictive versus liberal fluid management is underway. Inhaled nitric oxide has been used to improve oxygenation but has not resulted in any outcome benefit. Glucocorticoids may be beneficial in the fibroproliferative phase of lung injury by suppressing chronic inflammation. Rigorous clinical trials of new and established interventions are required to determine optimum therapy and reduce mortality in ALI/ARDS. PMID- 11867870 TI - Fluid resuscitation: colloids vs. crystalloids. AB - The administration of intravenous fluids is perhaps the most common treatment given in the intensive care unit. According to biologic rationale, ongoing fluid losses should be replaced to maintain fluid homeostasis and relative or absolute deficiencies in circulating blood volume should be prevented or rapidly corrected. There is agreement that insensible fluid losses and isotonic fluid losses should be replaced with a judicious mixture of water and crystalloid solutions. There is, however, a great deal of controversy on which fluids should be used during the acute resuscitation phase to deal with acute relative or absolute hypovolemia. In particular, there is much controversy on whether colloidal or crystalloid solutions should be used. The controversy remains despite multiple meta-analyses. There are suggestions that albumin might increase mortality but the data supporting these concerns are weak. Starch solutions might adversely affect renal function. Crystalloids might be favored in trauma patients. These views remain inadequately supported by evidence. A randomized controlled trial now under way should increase the evidence base for practice in this area. PMID- 11867871 TI - Vasopressors and the kidney. AB - The changes in renal perfusion induced by vasopressors depend on their effects on systemic hemodynamics and renal vascular resistance. Both effects are largely influenced by the patient's underlying condition such as myocardial contractility and vascular responsiveness. A beneficial effect can be expected if mean arterial pressure increases without decreasing cardiac output and if the effect on renal vascular resistance is less pronounced than on systemic vascular resistance. Acute renal failure is associated with loss of renal autoregulation and sepsis is associated with blood pressures below the autoregulatory threshold. Both conditions might therefore benefit from the administration of vasopressors. Many experimental and clinical data indeed suggest a beneficial effect of norepinephrine on the urine output in sepsis. A beneficial effect on renal function (glomerular filtration) is a less consistent finding suggesting that pressure diuresis might be partially responsible for the pressor-induced diuresis. Administration of vasopressors to patients with oliguria should be considered in fluid-resuscitated patients with distributive shock. Whether other vasopressors offer advantages over norepinephrine requires further investigation. PMID- 11867868 TI - Association between ACE gene polymorphism and diabetic nephropathy in South Indian patients. AB - OBJECTIVE: To study the association of ACE gene polymorphism and diabetic nephropathy in South Indian subjects. SETTING: Outpatient clinic of a specialized hospital. PATIENTS: The study included 109 South Indian type 2 diabetic patients (72 males and 37 females; age 56.7 plus/minus 9.0 years, mean plus/minus SD). The patients were subdivided into two groups: nephropathic (n=86) and normoalbuminuric patients (n=23). INTERVENTIONS: Genomic DNA was isolated from the peripheral blood leukocytes. To determine the ACE genotype, genomic DNA was amplified by PCR initially using a flanking primer pair and, subsequently when necessary, with a primer pair that recognizes the insertion specific sequence for confirmation of the specificity of the amplification reactions. MAIN OUTCOME MEASURES: ACE genotype distribution in the two study groups. RESULTS: In the nephropathic patients, ID and DD genotypes were present in 52.3% and 27.9% of the patients, respectively as compared to 34.8% and 21.7% respectively in those with normoalbuminuria. The D allele was present in 80.2% of the nephropathic patients and 56.5% of the normoalbuminuric patients (chi-squared=4.28, P=0.039; odds ratio 3.12). Therefore, the higher percentage of II genotype in the normoalbuminuric group was 43.5% as compared to the 19.8% in nephropathic patients. CONCLUSIONS: This study showed a positive association between the D allele (ID and DD genotype) of the ACE polymorphism and diabetic proteinuria in South Indian type 2 diabetic patients. Our findings are in keeping with several earlier studies showing a strong association of the D allele of the ACE gene with diabetic nephropathy. PMID- 11867872 TI - Acute-on-chronic liver failure: pathophysiological basis of therapeutic options. AB - The vast majority of patients that are referred to a specialist hepatological centre suffer from acute deterioration of their chronic liver disease. Yet, this entity of acute-on-chronic liver failure remains poorly defined. With the emergence of newer liver support strategies, it has become necessary to define this entity, its pathophysiology and the short- and long-term prognosis. This review focusses upon how a precipitant such as an episode of gastrointestinal bleeding or sepsis may start a cascade of events that culminate in end-organ dysfunction and liver failure. We briefly review the pathophysiological basis of the therapeutic modalities that are available. Our current strategy for the management of liver failure involves supportive therapy for the end-organs with the hope that liver function would recover if sufficient time for such a recovery is allowed. Because liver failure, whether of the acute or acute-on-chronic variety, is potentially reversible, the stage is set for the application of newer liver-support strategies to enhance the recovery process. PMID- 11867873 TI - Acid-base and electrolyte management in continuous renal replacement therapy. AB - Continuous renal replacement techniques are often utilized to manage acid-base and electrolyte problems in the critically ill patient. These techniques have an inherent capacity to manipulate the plasma composition and can be utilized efficiently to maintain homeostasis and metabolic control. Unfortunately, the efficacy of these techniques also permits wide variation in their use and can result in complications if they are not used appropriately. In most instances complications can be prevented by recognition of the operating principles and careful attention to detail. This article provides an overview of the principles of acid-base and electrolyte management with continuous renal replacement therapy. PMID- 11867874 TI - Continuous renal replacement therapy. Keeping pace with changes in technology and technique. AB - The rapidly changing nature of new technologies and techniques in acute health care means it can be difficult keeping pace. Most facilities, large or small, are usually in continuous evaluation of a new technology. Published reviews and professional group guidelines can assist the process of change for continuous renal replacement therapy (CRRT) technologies and techniques. The current techniques and technologies are a mixed application of old and new technologies providing a combination of convective and diffusive solute clearance methods. There are a variety of anticoagulation approaches. New, purpose-built CRRT machines offer many advantages over old technology but their costs can be prohibitive and users do not always meet them with rapid behavioral change. Reading journal publications and texts, scientific meetings, education and training, Internet web site review/participation, quality improvement activities and an accurate local data base are the keys to keeping pace with changes and identifying whether a benefit can be anticipated and demonstrated. Possible changes for the future of techniques and technologies may be in the areas of modified approaches to continuous therapy with tailored approaches for specific patient care settings. Improved membrane characteristics for wider indications and the bio-artificial kidney are emerging along with blood pump and circuit design improvements, with new machine/operator interfaces. PMID- 11867875 TI - Continuous renal replacement therapy. Keeping the circuit open: lessons from the lab. AB - BACKGROUND: Hemofiltration circuits generally require anticoagulation to prevent the membrane from clotting. Understanding the mechanisms involved in premature clotting of the filtration circuit is useful to optimize anticoagulation and maintain filter patency. AIMS: To discuss research performed at our institution which throws light on the causes of premature clotting of the hemofilter, and to highlight our approach to anticoagulation. DISCUSSION: Premature clotting of the circuit is related to low baseline levels of antithrombin III (AT-III), heparin co-factor II and tissue factor pathway inhibitor. Clotting is preceded by a precipitous rise in thrombin-antithrombin complexes, suggesting thrombin generation. Our standard anticoagulant is unfractionated heparin, where the primary mode of action is potentiation of endogenous AT-III. However, AT-III levels are diminished in sepsis and other causes of systemic inflammation. To supplement or prevent consumption of AT-III we use fresh frozen plasma or aprotinin (as cheaper alternatives to AT-III) in situations where filter life span is limited and mechanical obstruction has been excluded. Anticoagulation can be managed in heparin-induced thrombocytopenic patients with danaparoid, prostaglandins and/or predilution. PMID- 11867876 TI - A review of plasma exchange in sepsis. AB - BACKGROUND: Severe sepsis involves a generalised inflammatory response, mediated by a number of cellular and humoral factors. Modulation of this response holds the promise of improved survival. Plasma exchange has been suggested as an adjunctive therapy in grave infective illnesses such as meningococcaemia, because it might remove harmful bacterial products and excessive endogenous inflammatory mediators. AIMS: The aim of this article is to outline plasma exchange as an adjunctive therapy in sepsis, with an emphasis on the available clinical and experimental evidence for its use. METHODS: A literature review was performed using Medline including all English language references to exchange transfusion, plasma exchange, plasmapheresis, plasma filtration and sepsis. Relevant texts and conference proceedings booklets were also hand searched. RESULTS: Uncontrolled human data from case reports of more than 40 patients treated with plasma exchange for severe sepsis suggest a survival rate of over 70%. Animal studies produced conflicting results depending on the species and the model of sepsis employed. The only controlled clinical trial was too small to make conclusions regarding mortality. CONCLUSIONS: Plasma exchange remains an intuitively attractive but unproven therapy in sepsis. More controlled clinical trials are needed. PMID- 11867878 TI - Acute renal failure after cardiothoracic surgery: a review of three years experience. AB - 2,683 cardiothoracic operations were carried out over a 3-year period. Patients requiring haemofiltration after surgery had a much greater mortality than those not haemofiltered. Of the 1,177 cardiothoracic intensive care unit (ICU) patients, 91 required haemofiltration for acute renal failure (ARF; 7.7%). Of the 1,506 cardiothoracic high-dependency unit patients 13 were transferred to the renal unit for dialysis (0.86%). Mortality for cardiothoracic patients overall was 14.4% and for those who required haemofiltration 58.7%. 74 of these haemofiltered patients had normal renal function preoperatively; mortality 61%. 15 patients had pre-existing renal impairment; mortality 53.3%. 15 patients were on dialysis prior to surgery; mortality 60%. Age was not a predictor of requirement for renal replacement therapy or of mortality. Operation type was a risk factor for ARF: of patients having a coronary artery bypass graft (CABG) 2.4% were filtered (mortality 37.8%), of patients having valve replacements 14.2% were haemofiltered (mortality 60.9%), and of the patients having redo-CABGs or redo-valve replacements 12.3% required haemofiltration (mortality 100%). PMID- 11867877 TI - Coupled plasma filtration adsorption. AB - Both pro-inflammatory and anti-inflammatory mediators participate in the pathogenesis of sepsis and explain the failure of specific therapies to improve survival. Continuous extracorporeal therapies have been proposed as a therapeutic option in sepsis. We have studied the effects of plasma filtration associated with adsorption in patients with septic shock. We have shown that such treatment may lead to improved survival in a rabbit model of sepsis and to improved hemodynamics, reduced norepinephrine dose and restoration of near-to-normal responsiveness of blood leukocytes to endotoxin in humans. It is anticipated that treatment of plasma, as a device modular to conventional hemofiltration, may pave the way to innovative approaches to the extracorporeal treatment of septic patients. PMID- 11867879 TI - How to feed patients with renal dysfunction. AB - Renal dysfunction is common in critically ill patients and its presence has, in the past, posed serious challenges to nutritional support. Such challenges were due to the increased azotemia induced by protein or amino acid administration, the fluid overload caused by the administration of nutrients and the difficulties associated with the control of these complications by means of conventional dialytic techniques. The development and increasing application of continuous renal replacement therapy (CRRT) has removed such concerns, because control of azotemia and fluid balance can be predictably and reliably achieved in all patients. Accordingly, the presence of renal failure should in no way influence the amount or type of nutritional support administered to a critically ill patient. We recommend that approximately 30-35 kcal/kg/day be administered enterally and begun within the first few hours of admission to the intensive care unit and that protein intake be kept in the 1.5-2 g/kg/day range. Accumulating evidence also suggests that immune-enhancing enteral preparations decrease the duration of hospital stay, the number of infections and perhaps mortality. Such preparations should be used in these patients. Finally adequate vitamin and trace element supplementation is recommended to counterbalance the decrease in antioxidants and the loss of some vitamins during CRRT. Available evidence suggests that if these steps are applied as part of a protocol-based approach to the nutritional support of patients with renal failure, their morbidity and perhaps mortality can be significantly decreased. PMID- 11867881 TI - Hyperglycemia in patients with focal cerebral ischemia after intravenous thrombolysis: influence on clinical outcome and infarct size. AB - The aim of the present prospective study was to investigate whether hyperglycemia influences the clinical outcome or the infarct size after intravenous thrombolysis of focal cerebral ischemia. A consecutive series of hyperglycemic (n = 14) and normoglycemic patients (n = 17) with acute focal cerebral ischemia (<3 h) in the middle cerebral artery (MCA) territory received rtPA (0.9 mg/kg body weight) intravenously. Clinical outcome was measured using the NIH Stroke Score on admission and was followed up until day 28. Infarct volume was measured by diffusion-weighted MR imaging on admission, on days 3 and 7. There was a significantly better neurological outcome on day 28 in the normoglycemic patients than in the hyperglycemic group (NIH SS 4.0 versus 7.4; p < 0.05). The infarction volume increased significantly in the hyperglycemic patients Delta = 39.9 plus minus 17.4% compared to normoglycemic patients Delta = 27.1 plus minus 14.1% (p < 0.05). The present study suggests that hyperglycemia in patients with a focal MCA ischemia can cause a worse clinical outcome despite recanalization of the occluded vessel by thrombolysis therapy. This correlates with a markedly larger increase of the infarction volume in the hyperglycemic group. These results may be explained by an accentuated lactate accumulation and pH decrease by elevated energy levels which cannot be compensated by restoration of blood flow alone. PMID- 11867880 TI - Intracranial dural arteriovenous fistulas: analysis of 60 patients. AB - OBJECTIVE: To analyze and update the clinical symptomatology, CT and MRI findings, angiographic features, and therapeutic outcomes of patients with dural arteriovenous fistulas (DAVFs). BACKGROUND: Studies of DAVFs in a large number of patients have seldom appeared in neurology literature. METHODS: The authors investigated 60 consecutive patients with DAVFs who were admitted between January 1991 and January 2001. The DAVFs were graded into five types according to the classification of Cognard et al. [Radiology 1995;194:671-680]. Presumable etiologies, clinical features, imaging findings and therapeutic outcomes were evaluated on the basis of the location and type of DAVFs. RESULTS: Sinus thrombosis, previous trauma, craniotomy, cerebral infarction and acupuncture were detected as possible etiologic factors of DAVFs. The cavernous sinus (57%) was the most common location of DAVFs. Although the neurological symptoms were closely related to the location of the DAVFs, in some patients, there were also symptoms that did not reflect the location. Although the women outnumbered the men, the men presented with aggressive neurological manifestations more often (p < 0.05). Ten out of 12 patients (83%) with DAVFs involving locations other than the large sinuses presented with aggressive neurological manifestations. 70% of brain CTs and 81% of brain MRIs showed abnormal findings suggestive of DAVFs. Of 33 patients who underwent only endovascular embolization, 29 patients (88%) were cured or improved. Radiosurgery and surgical excision done in some patients provided fair results. Patients with DAVFs involving large sinuses showed a better therapeutic outcome than those in whom locations other than the large sinuses were involved, while patients without venous ectasia had more a favorable outcome than those with it (each, p < 0.01). CONCLUSIONS: DAVFs result from various etiologic factors, show diverse manifestations usually reflecting the location and can be treated successfully in most patients. Factors related to poor clinical outcome include male sex, the presence of venous ectasia and involved locations other than the large sinuses. PMID- 11867883 TI - Patent foramen ovale, a possible cause of symptomatic migraine: a study of 74 patients with acute ischemic stroke. AB - Recent studies reported an increased prevalence of patent foramen ovale (PFO) in patients with migraine with aura (MA+). To investigate the possible relationship between MA+ and paradoxical embolism, we studied the prevalence of both conditions. Investigation of PFO was undertaken in 74 consecutive patients presenting with an acute stroke of undetermined origin. The patients were questioned about MA+ or migraine without aura (MA-) according to the criteria of the International Headache Society. Follow-up was performed to investigate the evolution of MA+ and MA- according to different treatments of stroke. PFO was found in 44 of 74 patients, 16 of whom had MA+ (36%), compared to 4 (13%) MA+ patients without PFO (p = 0.03). Of 25 patients in whom the PFO was considered to play a causal role in the stroke, 13 (52%) had MA+, whereas only 3 (16%) of 19 patients in whom PFO was considered unrelated had MA+ (p = 0.014). Thirty-nine of the patients with MA+ and MA- were studied over a mean follow-up of 13 months. Seven of 15 patients with MA+ and PFO, treated either with surgical closure or anticoagulants, noticed complete disappearance of MA+ attacks. The prevalence of MA+ is high among stroke patients with PFO. In patients with a high presumption of paradoxical embolism, the proportion of MA+ is increased, and this suggests a possible role of this association in the occurrence of the cerebrovascular event. Forty-seven percent of patients with PFO and MA+ reported complete suppression of their aura attacks after surgical closure or anticoagulant treatment. This finding suggests that at least in some patients, MA+ attacks may be due to paradoxical embolism. PMID- 11867882 TI - Effect of acute glycaemic index on clinical outcome after acute stroke. AB - Studies have shown that hyperglycaemia acutely after stroke independently predicts poorer survival and independence. Whether the change in glycaemic index in the acute phase of stroke has any effect on stroke outcome is unclear. Glycated serum proteins (GSP) reflect blood glucose concentration during the preceding 2 weeks. The aim of this study is to measure the association between the change in GSP % in the first 2 weeks after stroke and outcome. 167 patients were included. 117 (70%) patients were alive at 3 months. Admission glucose was higher in dead patients (7.8 mmol/l) compared to survivors (6.6 mmol/l) (p < 0.01). GSP at day 14 was higher in non survivors (21.8%) compared with survivors (19.1%) (p < 0.0001) as was the change in GSP (2.0 %) in non survivors compared with survivors (0.1%) (p < 0.0001). After adjusting for case mix, the change in GSP % was significantly associated with stroke mortality (p = 0.04). The odds ratio for death at 3 months after stroke associated with every 1% increase in change between GSP day 14 and GSP day 0, was 1.28 (95% CI: 1.1-1.62). Increases in glycaemic index as determined by GSP % are associated with excess in stroke mortality after adjusting for case mix. Intervention trials are required to test the hypothesis that improving glycaemic index after acute stroke improves outcome. PMID- 11867884 TI - Angioplasty and stenting for total symptomatic atherosclerotic occlusion of the subclavian or innominate arteries. AB - Bypass grafting is considered the treatment of choice for occlusions of supraaortic vessels. Percutaneous transluminal angioplasty (PTA) is a well established treatment for patients with subclavian stenosis, but its efficacy in total subclavian or innominate occlusions is so far not well established. Since 1995, we have carried out 9 PTA revascularizations in 9 patients with total symptomatic atherosclerotic occlusion of supraaortic vessels. The technical success rate was 100%, without any major (death or stroke) or minor complications. In the follow-up period (mean 37.4 months), 1 patient had a symptomatic left internal carotid artery occlusion and an asymptomatic occlusion of the treated subclavian artery. In expert hands, stenting-PTA should be considered as the primary therapeutic option. PMID- 11867885 TI - Carotid angioplasty with cerebral protection and stenting: report of 164 patients (194 carotid percutaneous transluminal angioplasties). AB - BACKGROUND: The introduction of cerebral protection devices with systematic stent placement has changed the nature of carotid artery stenosis treatment, reducing the immediate periprocedural complications and delayed restenosis. METHODS: We treated 164 patients with 194 carotid artery stenosis procedures; 92% of them were symptomatic patients. RESULTS: The morbidity rate of our series was 1.03% and the mortality was 1.9%. CONCLUSIONS: In the future, carotid stenosis treatment should perhaps be performed as a preventative measure and not used as a cure for full-blown symptoms. This could be effective in reducing the morbidity and mortality rates of this pathology. PMID- 11867886 TI - The VITATOPS (Vitamins to Prevent Stroke) Trial: rationale and design of an international, large, simple, randomised trial of homocysteine-lowering multivitamin therapy in patients with recent transient ischaemic attack or stroke. AB - BACKGROUND: Epidemiological studies suggest that raised plasma concentrations of total homocysteine (tHcy) may be a common, causal and treatable risk factor for atherothromboembolic ischaemic stroke. Although tHcy can be lowered effectively with small doses of folic acid, vitamin B(12) and vitamin B(6), it is not known whether lowering tHcy, by means of multivitamin therapy, can prevent stroke and other major atherothromboembolic vascular events. PURPOSE: To determine whether vitamin supplements (folic acid 2 mg, B(6) 25 mg, B(12) 500 mug) reduce the risk of stroke, and other serious vascular events, in patients with recent stroke or transient ischaemic attacks of the brain or eye (TIA). METHODS: An international, multi-centre, randomised, double-blind, placebo-controlled clinical trial. RESULTS: As of November 2001, more than 1,400 patients have been randomised from 10 countries in four continents. CONCLUSION: VITATOPS aims to recruit and follow up 8,000 patients between 2000 and 2004, and provide a reliable estimate of the safety and effectiveness of dietary supplementation with folic acid, vitamin B(12), and vitamin B(6) in reducing recurrent serious vascular events among a wide range of patients with TIA and stroke. PMID- 11867887 TI - Stroke in hospitalized patients: are there special risk factors? AB - BACKGROUND AND PURPOSE: The occurrence of stroke in patients hospitalized for various illnesses remains a particular challenge for neurologists. Determining the potential causes for these particular cerebrovascular events may help to define the population at risk and to take measures in order to prevent stroke during hospitalization. The aim of our study was to evaluate the potential risk factors associated with stroke, which occurred in patients hospitalized for other illnesses. METHODS: This retrospective case-control study based on data of patients who underwent an ischemic stroke while being hospitalized not because of stroke and a control group of patients admitted during the same period who were matched for age and sex to the study patients. Common vascular risk factors, e.g. fever, leukocytosis, blood pressure, hemoglobin, cardiac arrhythmia and dehydration, were compared between the study and control groups. RESULTS: Of 2,247 consecutive patients with ischemic stroke, the stroke had occurred during hospitalization not related to any surgical procedure in 80 (3.5%). Six parameters were found as being significant independent risk factors for in hospital stroke: fever and leukocytosis during hospitalization, elevated diastolic and unstable blood pressure, dehydration and past history of myocardial infarction. CONCLUSION: Careful monitoring of temperature, blood count, blood pressure, and clinical and laboratory signs of dehydration is needed to prevent in-hospital stroke. PMID- 11867888 TI - Socioeconomic aspects of postacute care for patients with brain infarction in France. AB - BACKGROUND: In France, the socioeconomic aspects of stroke have never been addressed. Such analyses are essential for health authorities to justify the establishment of new stroke units when resources are low, provided it can be shown that stroke units are effective in reducing both the morbidity and mortality of stroke. Only 6 dedicated stroke services exist for 60 million inhabitants in France. Our aim was to study acute and postacute pathways and to determine the factors that influence destination after discharge, handicap evolution and costs. METHODS: In a cohort of 494 consecutive patients with brain infarction, we collected information on medical and socioeconomic variables, handicap and its evolution using the modified Rankin scale and Mini-Mental Status score at the 10th day, 6th month and 18th-40th month. These data were recorded during the initial hospital stay, at the follow-up clinic visit and in a home interview done 18-40 months after discharge by research nurses. We used multiple logistic regression for analyses. RESULTS: The most important factor for not returning home was having a Rankin score greater than 3 with an odds ratio of 41.7 (95% confidence interval 19.2-90.0; p = 0.001). Multivariate analysis showed that when the Rankin score was 0, 1 or 2, the main factors for not returning home were socioeconomic variables and serious medical disorders. When the Rankin score was 4 or 5, the main reason for not being sent for rehabilitation was medical status. After adjustment for the Rankin score, patients who returned home or were transferred to rehabilitation were quite similar regarding socioeconomic and medical variables. Other patients transferred to a geriatric ward, nursing home or new housing were more frequently living alone, 60 years of age or older, had less than 2 children, low level of education, dementia or cancer. Overall, the mean cost was 19,513 Euros over an 18-month period and was mainly driven by the level of the Rankin score (e.g. 10,530 vs. 34,809 Euros for Rankin scores of 0-1 and 4-5, respectively). CONCLUSION: These data showed that not only handicap level but also socioeconomic variables are important in determining the destination of stroke patients after discharge. They may help health authorities to make decisions to establish new approaches to treat stroke. This study can also serve as a basis for future cost-effectiveness studies of new drugs being evaluated in therapeutic trials or of new management strategies of stroke patients. PMID- 11867890 TI - A rare case of stroke in Wegener's granulomatosis. PMID- 11867889 TI - Aortic embolism in Claude's syndrome. PMID- 11867891 TI - Isolated facial paresis due to a centrum ovale infarction. PMID- 11867892 TI - Scalp necrosis--a rare complication of temporal arteritis. PMID- 11867893 TI - Surgical research--a discipline at the interface of the basic sciences and the clinic. Festschrift dedicated to Konrad Messmer on the occasion of his 65th birthday. PMID- 11867895 TI - The role of research in the training of residents. PMID- 11867896 TI - Surgical research and Helicobacter pylori infection -- a contradiction? PMID- 11867894 TI - Surgical research and clinical routine in the new century. PMID- 11867898 TI - Twenty years of shock wave research at the Institute for Surgical Research. PMID- 11867897 TI - Quality of life in animals as a new outcome for surgical research: G-CSF as a quality of life improving factor. AB - Sepsis is still a major problem in human medicine with a high mortality rate. Nearly all attempts to improve the outcome of septic patients with immune modulators failed. In most of these trials only mechanistic endpoints such as mortality rate, complication rate, cytokine levels and physiological parameters were assessed. Only in a very few trials quality of life had been chosen as primary endpoint. In basic research and especially in animal experiments in the field of sepsis and oncology, only molecular investigations which explain drug and treatment interactions were in the focus of the scientific community. Animal models simulating clinical complexity and investigating outcomes like quality of life were very rare. The aim of this study was to demonstrate alterations in sickness behaviour -- the animal equivalent to quality of life in man -- in rats as a response to sepsis after prophylaxis with G-CSF and antibiotics. Sickness behaviour was assessed by measurement of core body temperature, food and water intake, locomotor activity and circadian rhythm of these parameters. Complex animal experiments in rats were performed including anaesthesia, antibiotic and cytokine (G-CSF) prophylaxis, volume substitution, laparotomy, contamination and infection with human faecal suspension and postoperative analgesia. In group A (sham) and D (antibiotic + G-CSF) the mortality rate was 0%, but in group B (no prophylaxis) 33% (3/9) and in group C (antibiotic prophylaxis) 11% (1/9) of the animals died. Before infection all rats showed clear circadian patterns of locomotor activity and body temperature with physiologically higher values during the night-time. Immediately after operation and infection temperature increased, water and food intake, locomotor activity decreased and circadian rhythms were lost. Body temperature and water consumption were already normalised at day 2 after infection in all groups. Normal food intake was re-established in group C and D at day 3 while one more day was needed for recovery in Group C. Restoration of locomotor activity occurred in group D at day 5, in group C at day 7. In group B locomotor activity remained suppressed during the whole observation period of 8 days postoperatively. In conclusion, in septic rats sickness behaviour, an equivalent to quality of life in humans, is improved rapidly by a prophylaxis with G-CSF in combination with antibiotics and can be used as a new outcome in preclinical surgical research. PMID- 11867899 TI - Function of BID -- a molecule of the bcl-2 family -- in ischemic cell death in the brain. AB - Mitochondrial mechanisms, particularly the release of cytochrome c, play a role in the death of nerve and glial cells in cerebral ischemia. We have currently investigated whether BID, a proapoptotic molecule of the bcl-2 family and promoter of the release of cytochrome c is expressed in the brain, activated by cerebral ischemia in vivo, and contributes to ischemic cell death. We found BID in the cytosol of mouse brain and of primary cultured mouse neurons and showed that neuronal BID is a substrate for caspase 8. BID was cleaved in vivo 4 h after transitory occlusion of the middle cerebral artery. Further, BID(-/-) mice had a significant attenuation of infarction (-67%) and significantly lower release of cytochrome c (-41%). The findings indicate that the proapoptotic molecule BID may contribute to the demise of nerve cells from cerebral ischemia by release of cytochrome c and activation of caspase. PMID- 11867900 TI - Prospective system analysis of the pre- and early hospital care in severe head injury in Bavaria at a population-based level. PMID- 11867901 TI - Response of alveolar macrophages to inhaled particulates. PMID- 11867902 TI - L-arginine: a unique amino acid for improving depressed wound immune function following hemorrhage. AB - OBJECTIVE: To determine whether L-arginine has any salutary effects on wound immune cell function following trauma-hemorrhage. BACKGROUND: Depressed wound immune function contributes to an increased incidence of wound infections following hemorrhage. Although administration of L-arginine has been shown to restore depressed cell-mediated immune responses following hemorrhage potentially by maintaining organ blood flow, it remains unknown whether L-arginine has any salutary effects on the depressed local immune response at the wound site. METHODS: Male mice were subjected to a midline laparotomy and polyvinyl sponges were implanted subcutaneously in the abdominal wound prior to hemorrhage (35 +/- 5 mm Hg for 90 min and resuscitation) or sham operation. During resuscitation mice received 300 mg/kg body weight L-arginine or saline (vehicle). Sponges were harvested 24 h thereafter, wound fluid collected and wound immune cells cultured for 24 h in the presence of LPS. Pro- (IL-1 beta, IL-6) and anti-inflammatory (IL 10) cytokines were determined in the supernatants and the wound fluid. In addition, wounds were stained for IL-6 immunohistochemically. In a separate set of animals, skin and muscle blood flow was determined by microspheres. RESULTS: The capacity of wound immune cells to release IL-1 beta and IL-6 in vitro was significantly depressed in hemorrhaged mice receiving vehicle. Administration of L-arginine, however, improved wound immune cell function. In contrast, in vivo the increased IL-6 release at the wound site was decreased in L-arginine-treated mice following hemorrhage. Moreover, IL-10 levels were significantly increased in the wound fluid in hemorrhaged animals receiving L-arginine compared to vehicle treated mice. In addition, the depressed skin and muscle blood flow after hemorrhage was restored by L-arginine. CONCLUSIONS: Thus, L-arginine might improve local wound cell function by decreasing the inflammatory response at the wound site. Since L-arginine protected wound immune cell function this amino acid might represent a novel and useful adjunct to fluid resuscitation for decreasing wound complications following hemorrhage. PMID- 11867903 TI - Chronic porcine two-hit model with hemorrhagic shock and Pseudomonas aeruginosa sepsis. AB - BACKGROUND: Sepsis is still a major cause of death despite well-developed therapeutical strategies such as antibiotics and supportive medication. The aim of this study was to characterize the long-term effects of a two-hit porcine sepsis model with a hemorrhagic shock as 'first hit' followed by a Pseudomonas aeruginosa infusion as 'second hit'. MATERIALS AND METHODS: Twelve juvenile healthy pigs were anesthetized and hemodynamically monitored. The two-hit group (n = 6) underwent a hemorrhagic shock with a 50% reduction of the mean arterial pressure and/or cardiac index for 45 min, followed by resuscitation, while the control group (n = 6) received no pretreatment. All chronically catheterized conscious pigs were challenged with a P. aeruginosa infusion (1.6 x 10(7) CFU/kg/h for the first 24 h followed by 1.6 x 10(6) CFU/kg/h for the next 24 h) and observed for another 48 h. RESULTS: The two-hit group showed the following significant differences to the control group: higher APACHE II scores prior to sepsis induction, increased persisting mean pulmonary arterial pressure (MPAP) and pulmonary vascular resistance index (PVRI) during bacterial challenge. In contrast, systemic vascular resistance (SVRI) was reduced at the end of the study. Throughout the observation period, the mean arterial pressure (MAP) was significantly reduced. CONCLUSIONS: The present study shows that the clinical course and hemodynamic effects of a P. aeruginosa sepsis will be aggravated by a preceding hemorrhagic shock during an observation period of 96 h. This two-hit model represents a valid, clinically relevant experimental protocol in sepsis research. PMID- 11867904 TI - Endotoxin inactivation by enterally applied colostrum of different composition. AB - BACKGROUND AND PURPOSE: Enteral applied bovine colostrum can significantly reduce endotoxin concentration in plasma. Since colostrum is a mixture of biological active ingredients 3 possible substances which are able to influence the endotoxin elimination were concentrated in 3 different colostrum products. Immunoglobulin-, lactoferrin- and casein-enriched colostra and lactoferrin alone were orally administered to endotoxinaemic rats. METHODS: Endotoxinaemia was induced to rats by enteral application of 10(10) E. coli together with 40 mg Nebacetin. Control animals received albumin. From all rats plasma samples were taken over the time of 5 h and endotoxin concentration determined with limulus lysate and chromogenic substrate. RESULTS: Whereas in control animals as well as in animals treated with casein-enriched colostrum a marked increase of endotoxin values to over 130 EU/dl could be observed after 5 h, the oral application of gammaglobulin-enriched and especially lactoferrin-enriched colostrum decreased endotoxin values by more than 50%. The most effective endotoxin elimination was seen with lactoferrin alone. CONCLUSIONS: From this results it can be concluded that not only gammaglobulin but especially lactoferrin seems to be responsible for the elimination of endotoxin with regard to enterally applied colostrum preparations. PMID- 11867905 TI - Stentgrafts in the thoracic aorta -- the best application of a new idea -- but still in its beginning? PMID- 11867906 TI - Improvement of lung preservation -- from experiment to clinical practice. AB - BACKGROUND: Reperfusion injury represents a severe early complication following lung transplantation. Among the pathogenetic factors, the high potassium content of Euro-Collins(reg) solution is discussed. MATERIAL AND METHODS: In a pig model of orthotopic left-sided lung transplantation we investigated the effect of Euro Collins solution (EC: n = 6) versus low potassium dextran (LPD: Perfadex: n = 6). Sham-operated (n = 6) animals served as control. Transplant function, cellular energy metabolism and endothelial morphology served as parameters. In a clinical investigation, 124 patients were evaluated following single (EC: n = 31; LPD n = 37) or double (EC: n = 17; LPD n = 39) lung transplantation, whose organs where preserved with EC (n = 48) or LPD (n = 76). Duration of ischemia, duration of ventilation and stay on ICU were registered. Primary transplant function was evaluated according to AaDO(2) values. Cause of early death (30 days) was declared. RESULTS: Experimental results: After flush with EC and 18 h ischemia, a reduction of tissue ATP content (p < 0.01 vs inital value and LPD) was noted. Endothelial damage after ischemia was severe (p < 0.05 vs control), paO(2) was significantly decreased. CLINICAL RESULTS: In the LPD group, duration of ischemia was longer for the grafts transplanted first (SLTx and DLTx: p = 0.0009) as well as second (2. organ DLTx: p = 0.045). Primary transplant function was improved (day 0: SLTx: p = 0.0015; DLTx: p = 0.0095, both vs EC). Duration of ventilation and stay on ICU were shorter (n.s.). Reperfusion injury-associated death was reduced from 8% (EC) to 0 (LPD). CONCLUSION: In experimental lung preservation, LPD lead to an improved graft function. These results were confirmed in clinical lung transplantation. Clinical lung preservation, therefore, should be carried out by use of LPD. PMID- 11867908 TI - The marginated pool. AB - The pulmonary circulation harbors a large intravascular reservoir of leukocytes referred to as the Marginated Pool. This marginated pool is balanced by propelling and retaining forces acting on leukocytes during their passage through the pulmonary circulation. The present paper discusses these factors and their underlying mechanisms. PMID- 11867907 TI - Viewing the microcirculation through the window: some twenty years experience with the hamster dorsal skinfold chamber. AB - Intravital microscopy represents a sophisticated technique to study the microcirculation in health and disease. While most preparations used for those studies are acute in nature, the use of chamber preparations in the skinfold bear the advantage to allow for chronic studies with repeated analysis of the microcirculation over a prolonged period of time. The skinfold chamber model for microcirculatory analysis has been adapted to mice, rats and hamsters. Although the use of rats and, in particular, the use of mice has the advantage of the availability of species-specific tools, the use of the hamster as the experimental animal may be preferred due to anatomical reasons, which facilitate the microsurgical preparation and improve the quality of microscopic imaging. The use of the hamster dorsal skinfold chamber, firstly described by Endrich and coworkers in 1980, has brought out during the last two decades a considerable number of experimental studies within the fields of microcirculation physiology, inflammation and sepsis, ischemia-reperfusion, angiogenesis, and transplantation, indicating that the model has to be considered a versatile tool to study the microcirculation in health and disease. PMID- 11867909 TI - Blood and plasma viscosity and microvascular function in hemodilution. A perspective from La Jolla, California. PMID- 11867910 TI - Effects of ultra-purified polymerized bovine hemoglobin on local tissue oxygen tension in striated skin muscle -- an efficacy study in the hamster. AB - The development of hemoglobin-based oxygen carriers has been propagated for replacement of the oxygen carrying properties of red blood cells for almost one century. Using a Clark-type multi-wire oxygen surface electrode and the dorsal skin fold chamber model of the awake Syrian golden hamster, local tissue pO(2) was analyzed in the thin striated skin muscle before and after administration of an ultra-purified polymerized bovine hemoglobin solution (U-PBHb, Biopure Corp., Boston, Mass., USA) under the following experimental conditions: (a) hypervolemic infusion with U-PBHb at approximately 10% of calculated blood volume, and (b) isovolemic exchange transfusion with U-PBHb by replacing approximately 50% of calculated blood volume. Control animals of group a received equivalent treatment with either isotonic saline or dextran 60, control animals of group b received dextran 60. Local tissue pO(2) was found slightly decreased after both hypervolemic infusion and isovolemic exchange transfusion with U-PBHb, while frequency distribution curves of local tissue pO(2) were found more narrow (less values <10 mm Hg and >25 mm Hg), suggesting a more homogeneous tissue pO(2) distribution. The data thus indicate that U-PBHb slightly decreases mean tissue pO(2) after both hypervolemic infusion and isovolemic exchange transfusion which is accompanied by an effective homogenization of local tissue pO(2) distribution as compared to dextran 60. PMID- 11867911 TI - Realisation of a technique for the non-invasive, clinical assessment of microvascular parameters in man; the KM factor. PMID- 11867912 TI - Comparison of the new OPS imaging technique with intravital microscopy: analysis of the colon microcirculation. AB - BACKGROUND: The OPS imaging technique has been introduced for in vivo assessment of microcirculation in humans. The aim of this study was to validate the new technique against intravital fluorescence microscopy (IFM) for the visualization of colon microcirculation in a murine model of inflammatory bowel disease (IBD). METHOD: IBD was induced in Balb/c mice by dextran sulfate sodium, controls received normal water. In each animal, both the CYTOSCAN A/R and IFM were used to image the microcirculation (n = 7 in each group). The postcapillary venular diameter was analyzed on the colon muscularis and mucosa. RESULTS: The venular diameter correlated significantly between both methods representing the good correspondence between both methods. CONCLUSION: Our study demonstrates that the new technique for visualization of microcirculation without use of fluorescent dyes, the OPS imaging, allows for quantitative measurement of a key microcirculatory parameters of the mouse colon. PMID- 11867913 TI - Extracorporeal circulation and intestinal microcirculation: pathophysiology and therapeutical options. An intravital microscopic study in a large animal model. PMID- 11867914 TI - Small-volume resuscitation from hemorrhagic shock by hypertonic saline dextran -- conceptional basis and historical background. PMID- 11867915 TI - Different optical methods for clinical monitoring of the microcirculation. PMID- 11867917 TI - Tolerance: then and now. PMID- 11867916 TI - Munich and Cambridge -- a long collaboration in friendship and science. PMID- 11867918 TI - Postischemic reperfusion injury to allografts -- a case for 'innate immunity'? PMID- 11867919 TI - Adverse drug reactions: mechanisms and assessment. AB - Adverse drug reactions (ADR) are an important clinical problem. They account for about 5% of all hospital admissions and cause death in approximately 0.01% of surgical patients. The mechanisms leading to ADR beyond IgE-mediated allergy are still poorly understood. The importance of chemically reactive drug metabolites and the involvement of T-lymphocytes in many drug hypersensitivity reactions have been highlighted in recent years. ADR are diagnosed on clinical grounds and the temporal relation between drug intake and the appearance of the symptoms. Allergy tests are required in the further assessment of the reaction. By means of skin tests, in vitro tests and provocation tests information about the culprit drug, the mechanism involved and possible alternatives can be obtained. PMID- 11867920 TI - Chaos -- no randomness in cardiac physiology. PMID- 11867921 TI - Hyperoxia in extreme hemodilution. AB - Intraoperative surgical blood loss is initially replaced by infusion of red cell free, cristalloidal or colloidal solutions. When normovolemia is maintained the ensuing dilutional anemia is compensated by an increase of cardiac output and of arterial oxygen extraction. In the ideal case, a surgical blood loss can entirely be 'bridged' without transfusion by intraoperative normovolemic hemodilution. However major blood loss results in extreme hemodilution and the transfusion of red blood cells may finally become necessary to increase arterial oxygen content and to preserve tissue oxygenation. When transfusion has to be started before surgical control of bleeding has been achieved, parts of the red blood cells transfused will get lost, thereby increasing intraoperative transfusion needs. Beside red blood cell transfusion, arterial oxygen content can be rapidly increased by ventilating the patient with 100% oxygen (hyperoxic ventilation), thus enhancing the amount of physically dissolved oxygen in plasma (hyperoxia). In experimental and clinical studies hyperoxic ventilation has emerged as a simple, safe and effective intervention to enlarge the margin of safety for hemodynamic compensation and tissue oxygenation in hemodiluted subjects experiencing major bleeding. The hyperoxia-associated microcirculatory dysregulation and impaired tissue oxygenation known to take place in the presence of a physiologic hemoglobin concentration are not encountered in hemodiluted subjects. Hyperoxic hemodilution i.e. the combination of intraoperative extreme hemodilution and hyperoxic ventilation may therefore be considered a cost effective, safe and efficient supplement to reduce allogeneic transfusion during surgical interventions associated with high blood losses. The vast majority of the experimental and clinical investigations this new concept is based on was initiated and performed under the guidance of Prof. Konrad Messmer. PMID- 11867922 TI - Nephrotoxicity of halogenated inhalational anaesthetics: fictions and facts. PMID- 11867923 TI - Searching the ideal inhaled vasodilator: from nitric oxide to prostacyclin. AB - Today, the technique to directly administer vasodilators via the airway to treat pulmonary hypertension and to improve pulmonary gas exchange is widely accepted among clinicians. The flood of scientific work focussing on this new therapeutic concept had been initiated by a fundamental new observation by Pepke-Zaba [1]and Frostell in 1991 [2]: Both scientists reported, that inhalation of exogenous nitric oxide (NO) gas selectively dilates pulmonary vessels without a concomittant systemic vasodilation. No more than another decade ago NO was identified as an important endogenous vasodilator [3]while having merely been regarded an environmental pollutant before that time. Although inhaled NO proved to be efficacious, alternatives were sought-after due to NO's potential side effects. In search for the ideal inhaled vasodilator another group of endogenous mediators -- the prostanoids -- came into the focus of interest. The evidence for safety and efficacy of inhaled prostanoids is -- among a lot of other valuable work -- based on a series of experimental and clinical investigations that have been performed or designed at the Institute for Surgical Research under the guidance and mentorship of Prof. Dr. med. Dr. h.c. mult. K. Messmer [4-19]. In the following, the current and newly emerging clinical applications of inhaled prostanoids and the experimental data which they are based on, will be reviewed. PMID- 11867924 TI - Risk/benefit evaluation of drugs: the role of the pharmaceutical industry in Germany. AB - Drugs that are efficacious have usually also undesired side effects. When applying for marketing authorization of a drug, demonstration of a positive risk/benefit ratio is a prerequisite for approval by the competent authorities. Once on the market, risk/benefit evaluation has to be continued in order to determine whether the risk/benefit ratio is still positive or not. The German Drug Law, by means of article 5, poses this responsibility on the pharmaceutical entrepreneur. Specific instructions on how to perform the risk/benefit evaluation can be derived from article 5, and a decision matrix was developed to support taking action if the risk/benefit ratio has changed. Historical and current examples are presented to illustrate changes of the risk/benefit ratio, problems associated with detecting risk signals, and the resulting regulatory actions. PMID- 11867925 TI - Physicians and culture. PMID- 11867926 TI - The influence of Andalusia on German culture. PMID- 11867927 TI - Age-related bone loss: old bone, new facts. AB - The human skeleton serves several functions for the body: support, locomotion, protection of vital organs, and housing of bone marrow. Bone remodeling is the result of the interactions of multiple elements, including osteoblasts, osteoclasts, hormones, growth factors, and cytokines, the end result being the maintenance of the bone architecture and to maintain systemic calcium homeostasis. In early life, a careful balance exists between bone formation by osteoblasts and bone resorption by osteoclasts. With aging, the process of coupled bone formation is affected by the reduction of osteoblast differentiation, activity, and life span which is further potentiated in the perimenopausal years with hormone deprivation and increased osteoclast activity. Age-related bone loss is thus not only a consequence of hormone deprivation, but also the result of changes in bone formation and cell-cell interactions with a unique pathophysiology. In this review, we describe the cellular and metabolic changes associated with aging bone and present recent evidence regarding cell differentiation within the bone marrow. We also consider the mechanism of programmed cell death, apoptosis, as being an important determinant of aging in bone as well as describe possible future interventions to prolong the life span of osteoblasts. PMID- 11867928 TI - Nonhuman primate models of skeletal and reproductive aging. AB - BACKGROUND: Bone loss, osteoarthritis (OA) and age-related changes in reproduction are some of the most apparent and troubling results of advancing age in the human population. The significance of these changes underscores the need for developing appropriate animal models to study aging of bone and the reproductive system. OBJECTIVE: This review surveys the literature regarding the current state of nonhuman primates (NHP) as models for skeletal and reproductive aging in humans. RESULTS: Several species of NHP exhibit age-related bone loss as well as changes in the mineral content and architecture of bone that are similar to those reported in humans. In addition, since aged females of some species of NHP exhibit cessation of menses and serum hormone profiles consistent with those described in peri- and premenopausal women, they provide useful models of postmenopausal bone loss. Sensitivity to surgical estrogen depletion and hormone replacement has also been demonstrated in female NHP. Spontaneous development of degenerative arthritis, analogous to humans in age at onset, joint involvement and histologic changes, suggest that NHP are suitable for the study of human OA. Many similarities exist between female NHPs and women in age-related changes in reproductive function including hormone concentrations, menstrual cycling, fertility and the development of endometriosis. Changes in male sexual behavior and fertility with age are also comparable in men and male NHP. CONCLUSIONS: Genetic similarity, long lives and similar reproductive endocrinology suggest that NHPs are likely candidates as models of skeletal and reproductive aging in humans. Current data confirm that several species of NHP exhibit changes in bone and reproduction that are comparable to those known to occur in humans. PMID- 11867929 TI - Adenine nucleotide and creatine phosphate pool in adult and old rat heart during immobilization stress. AB - BACKGROUND: Disturbances in the heart energy provision is important in the stress induced injury of myocardium. In order to learn the causes of age-dependent differences in myocardial sensitivity to stress, we determined the level of adenylates, creatine and creatine phosphate in adult and old rat heart during an immobilization stress. METHODS: Fifty male Wistar rats were used to study the myocardial adenine nucleotides, creatine and creatine phosphate pool and creatine kinase activity during an immobilization period. RESULTS: The concentration of adenine nucleotides in old and adult rat hearts was similar. The total concentration of adenylate pool, ATP and ADP in the heart of both age groups was reduced during stress. However, in old rats under stress the concentration of ATP in a less measure and ADP in a greater measure is decreased against that observed in adult animals. The level of inorganic phosphate in old rat heart remains the same as in adult rats. CONCLUSION: The creatine kinase system may be important in stabilizing the ATP level in myocardium of adult rats during stress. The role of this system in heart energy metabolism during stress is decreased in old rats. Disturbances of isoenzyme creatine kinase activity are responsible for these disorders. PMID- 11867930 TI - Changes in serum cortisol with age in critically ill patients. AB - BACKGROUND: Mortality in the intensive care unit (ICU) rises with age, a high basal serum cortisol and a small response to adrenocorticotropin (ACTH) stimulation. Even slight impairment of the adrenal response during severe illness can be lethal. OBJECTIVES: To determine if age is associated with changes in basal or stimulated serum cortisol in critically ill patients. METHODS: We studied 2 groups of surgical ICU patients with hypotension despite > or = 6 h of catecholamine therapy. Group 1 comprised 7 patients aged <30 (mean 22.9 +/- 3.7) years, and group 2 comprised 8 patients aged >60 (mean 75.8 +/- 10.3) years (p < 0.001). We compared baseline serum cortisol levels and the serum cortisol response 30 and 60 min after stimulation with low-dose (1 microg) and 2 h later standard-dose (250 microg) ACTH. We also determined the incidence of adrenal insufficiency in each group using standard criteria and compared selected clinical variables. RESULTS: There was no significant difference in the mean serum cortisol at baseline although it tended to be higher in older patients. Group 2 patients had a significantly smaller response to the low-dose test at 30 min (p = 0.002), and to the standard-dose test at both 30 (p = 0.02) and 60 min (p = 0.04). There was no significant difference in the incidence of adrenal insufficiency between the 2 groups: 1/7 or 14.3% in group 1 vs. 1/8 or 12.5% in group 2 (p = 1.0). There was no significant difference between the 2 groups in the mean acute physiology score, blood pressure, serum albumin, dopamine, or dobutamine dose (p > 0.05). Creatinine clearance was significantly lower in group 2 (p > 0.001) and endogenous ACTH significantly higher (p = 0.04). Significantly more patients in group 1 (5/7, 72%) than group 2 (1/8, 12.5%) had a diagnosis of trauma on admission (p = 0.04). Seven of the eight patients (88%) in group 2 vs. 1/7 (14%) of patients in group 1 died in the ICU, but this difference was not statistically significant (p = 0.18). CONCLUSION: In this small exploratory study, baseline serum cortisol tended to be higher in older patients and older patients had a significantly smaller response to ACTH stimulation on both low dose and standard-dose tests. In view of the high death rate in the older ICU patients, the findings in this study need to be confirmed in a larger study. The mechanism and clinical significance of these findings remain to be determined but may be related to deterioration in renal function with age. PMID- 11867931 TI - Comprehension of written health care information in an affluent geriatric retirement community: use of the Test of Functional Health Literacy. AB - BACKGROUND: The comprehension of written health care information, e.g. prescription bottle labels or an informed consent form, impacts on communication with patients and their ability to understand medical procedures, as well as goals of therapy. OBJECTIVE: To determine if the problem of literacy was present in a population of affluent Albuquerque retirement community dwellers. METHODS: We administered the Test of Functional Health Literacy in Adults (TOFHL) to 93 residents (71 women and 22 men), using a one-on-one directed examination conducted by one of the authors (J.G.B.). RESULTS: The group studied had a mean age of >70 years, were not acutely ill, had no known loss of mental function, were better educated and had a high standard of living compared to national standards. However, 30% (95% confidence level) of our group were unable to adequately comprehend written information as measured by the TOFHL. The subgroup with poor comprehension (21 women and 7 men) had a mean age of 85 years (versus 82 years in the group with good comprehension) and a mean of 13 years (versus 15 years in the group with good comprehension) of formal education. Multivariate analysis was consistent with a negative effect of age (p = 0.04), a positive effect of years of formal education (p = 0.0005) and no effect of gender on the TOFHL score. CONCLUSION: Two recent studies, one in an emergency room setting and the other among Medicare enrollees, demonstrated similar results to ours. The high prevalence of poor comprehension in these groups should alert providers to the need to develop additional or different communication skills that can address the problem of poor reading comprehension across at least three diverse groups of people, with particular attention to those over the age of 65. PMID- 11867932 TI - Differential diagnosis of osteoporosis. AB - BACKGROUND: Dual x-ray absorptiometry (DEXA) is widely used to identify persons at increased risk for osteoporotic fractures. It does not, per se, give us any diagnostic information that is necessary to determine the best therapeutic approach in an individual case. OBJECTIVE: Identify conditions and tests that may impact on the diagnosis and the treatment of subjects with low bone densitometry scores. SUBJECTS AND METHODS: We evaluated clinically, by history, physical examination and laboratory tests, 272 persons with low spinal and/or femoral bone absorptiometry for potential causes of osteoporosis. RESULTS: We found that 25.3% of the patients had significant underlying conditions: 17.9% had subclinical vitamin D deficiency (osteomalacia), 6.7% had hypercalciuria and 0.7% had primary hyperparathyroidism. CONCLUSION: It is mandatory to go through a differential diagnosis in every case with absorptiometric data compatible with the diagnosis of osteoporosis. At the minimum, blood calcium and 25-hydroxyvitamin D and 24 hour urinary calcium excretion should be examined. This will allow the physician to determine the appropriate course of therapy in any individual patient who has abnormal DEXA findings. PMID- 11867933 TI - A strategy for the management of elderly women with primary hyperparathyroidism: a comparison of etidronate therapy with parathyroidectomy. AB - OBJECTIVE: To evaluate the efficacy of etidronate (EHDP) on lumbar spine bone mineral density (LSBMD) and total bone mineral density (TBMD) in elderly women with primary hyperparathyroidism (PHPT), we compared changes in LSBMD and TBMD between patients treated by EHDP therapy and parathyroidectomy (PTX). SUBJECTS AND METHODS: Twenty-two PHPT patients were enrolled and randomized into two groups; 9 received EHDP and 13 underwent PTX. All patients were followed up for 1 year by measuring LSBMD, TBMD, serum calcium, inorganic phosphate, parathyroid hormone, 1,25-dihydroxyvitamin D, serum alkaline phosphatase, intact osteocalcin, urinary pyridinoline (U(pyd)) and urinary deoxypyridinoline (U(dpd)). The presence of spinal fractures was evaluated by X-ray photography before and after treatment. RESULTS: EHDP treatment produced a significant increase in LSBMD of 10% compared with pretreatment levels after 1 year (p < 0.03, compared to baseline), while PTX produced a significant increase in LSBMD of 20% compared to pretreatment levels (p < 0.01). However, TBMD remained unchanged for 1 year after both EHDP administration and PTX. Among biochemical bone turnover markers, EHDP administration resulted in significant decreases in alkaline phosphatase by 78%, U(pyd) by 64% and U(dpd) by 37% after 12 months compared with the pretreatment levels (p < 0.05) and intact osteocalcin by 67% after 6 months (p < 0.05). There were no differences in the fracture rate between the EHDP and PTX groups during 1 year. CONCLUSION: EHDP administration results in a somewhat lower increase in LSBMD than that following PTX and suppresses bone formation and resorption in elderly PHPT patients for 1 year. We conclude that PTX is preferable to EHDP therapy for the management of elderly PHPT patients; however, EHDP administration should also be considered for elderly patients with many complications or who are unfit for surgery. PMID- 11867934 TI - Are stress resistance and longevity really linked in normal living conditions? AB - BACKGROUND: The existence of a positive relationship between stress resistance and longevity is often accepted, but this conclusion mainly stems from studies putting animals in adverse conditions such as food restriction. OBJECTIVE: It is argued that a positive correlation between stress resistance and longevity has not been clearly observed at the individual level in animals or humans living in nonstressful conditions. Experiments should verify this point. CONCLUSIONS: If experiments would fail to show this correlation, there would be no ground for the proposal that treatments stimulating stress resistance could increase longevity and health. PMID- 11867935 TI - Impact of age, gender and cognitive functioning on pain perception. AB - BACKGROUND: Experimental data on nociception in the elderly have so far been contradictory and most of these have been obtained using psychophysical methods with little attention paid simultaneously to the state of cognition and the psychometric performance of the subjects. OBJECTIVE: The aim of this study was twofold: (1) to evaluate the impact of age on experimental nociception thresholds, and (2) to investigate the interactions of age, sex, cognition and psychometric performance with nociception thresholds. METHODOLOGY: (1) Two groups, one young and one elderly, of 42 healthy participants each, 21 males and 21 females, were compared as regards nociception thresholds with thermal and mechanical stimuli (heat and pressure detection and tolerance thresholds). (2) The elderly group took cognitive (mini-mental test), psychometric (choice reaction time) and psychophysical tests in auditory (sensibility, tolerance, discrimination) and nociceptive fields. RESULTS: This study shows that (1) pressure nociception decreases with age especially in males, while thermal thresholds are not modified, and (2) correlations exist between cognitive function, psychometric performance, tolerance to loud sounds and tolerance to mechanical noxious stimuli. CONCLUSION: Our results suggest that cognitive and psychomotor parameters have to be taken into account when assessing experimental nociception in the elderly. Further studies are needed to evaluate possible report biases and to assess these interactions in old patients with pain and in mildly sensory, cognitive or motor impaired elderly subjects. PMID- 11867936 TI - Hypophyseal-portal somatostatin (SRIH) and jugular venous growth hormone secretion in the conscious unrestrained ewe. AB - Somatostatin (SRIH) release into hypophyseal portal blood varies reciprocally with growth hormone (GH) pulse generation in the male rat. However, few studies have directly evaluated this relationship in the female of any species. To address this issue, we carried out intensive (5 min) and extended (240 min) simultaneous monitoring of hypophyseal portal SRIH and internal jugular GH secretion in 7 unanesthetized ewes. Bihormonal synchrony was assessed by three statistically independent but complementary analyses: (i) cross-approximate entropy (X-ApEn) analysis to appraise the conditional regularity of SRIH/GH release patterns; (ii) cross-correlation analysis of paired sample SRIH and GH release rates, and (iii) probability analysis of random versus nonrandom SRIH and GH discrete pulse concordance. From a one-variable perspective, ApEn analysis documented consistently more irregular patterns of SRIH than GH release (94 +/- 4.3 and 72 +/- 8.1%, respectively, of the mean irregularity of 1,000 individual random-shuffled cognate series, p = 0.034). From a two-variable perspective, X ApEn analysis revealed a nearly mean random relationship between SRIH and GH release patterns (group mean +/- SEM, 94 +/- 4.5% of the mean asynchrony of 1,000 randomly shuffled SRIH/GH pairs). Cross-correlation analysis disclosed highly variable linkages between SRIH and GH secretion; viz, negative cross-correlations in 5 sheep, positive relationships in 4, and both positive and negative SRIH/GH associations in 2 animals, wherein changes in SRIH secretion either preceded or followed those of GH. Peak detection by model-free cluster analysis quantified a total of 28 SRIH and 31 GH release episodes. Corresponding interpulse intervals (min) were comparable (37 +/- 4 (SRIH) and 43 +/- 12 (GH)), but the mean fractional (%) amplitude of SRIH peaks was 3.5-fold lower (60 +/- 10%) than that for GH (225 +/- 50%) (p = 0.024). Pulse-concordance probability testing showed that discrete peaks of SRIH and GH secretion coincided only 33% of the time, although this value exceeded chance expectation (p < 10(-4)). In summary, the present analysis applies intensive (5 min) and extended (240 min) simultaneous sampling of hypophyseal-portal and jugular venous blood to quantitate the degree of coordinate SRIH and GH secretion in the unanesthetized ovariectomized ewe. Thereby, we unmask highly irregular SRIH release patterns, and nearly random SRIH and GH associations. We conclude that, to the extent that in vivo sampling reflects physiological SRIH/somatotrope activity, the female sheep maintains complex time-varying interactions between SRIH and GH release. PMID- 11867937 TI - Evidence for a central inhibitory role of growth hormone secretagogues and ghrelin on gastric acid secretion in conscious rats. AB - We examined the possible central and peripheral effects of synthetic growth hormone secretagogues (GHS), hexarelin (Hexa) and EP 40737 (D-Thr-D-Trp (2-Me) Ala- Trp-D-Phe-Lys-NH2), and of their endogenous counterpart, ghrelin, on gastric acid secretion. The compounds were administered intracerebroventricularly (i.c.v.) or subcutaneously (s.c.) in conscious male rats and the volume of gastric secretion and gastric acid output were examined 3 h after pylorus ligation (Shay-test). Central Hexa, EP 40737 and ghrelin administration (from 0.1 pmol to 1 nmol/rat, i.c.v.) significantly inhibited gastric acid secretion. The maximum inhibitory effect on gastric acid output was detected at the dose of 10 pmol/rat, i.c.v. for Hexa (-51.3%), of 100 pmol/rat, i.c.v. for EP 40737 (-70%) and of 1 pmol/rat, i.c.v. for ghrelin (-60%). All peptides were less effective at the highest dose used (1 nmol/rat, i.c.v.). Hexa, EP 40737 and ghrelin injected s.c. did not modify gastric acid secretion. The inhibitory action of Hexa on gastric acid secretion seems to involve brain somatostatinergic system since Hexa (10 pmol/rat, i.c.v.) did not inhibit gastric acid secretion in rats pretreated (4 h before) with cysteamine (300 mg/kg, s.c.), a depletor of endogenous somatostatin. These results show that synthetic GHS and ghrelin exert a central long-lasting inhibitory effect on gastric acid secretion in conscious pylorus ligated rats. The fact that very low doses of ghrelin and GHS inhibit gastric secretion, provide evidence for a tonic inhibitory role of the peptides in the central control of gastric secretory function. PMID- 11867938 TI - Direct visualization of internalization and intracellular trafficking of dopamine releasing protein-36aa. AB - Dopamine-releasing protein (DARP) is a potent regulator of dopamine (DA) release known to be involved in the development of rat catecholaminergic systems. In the present study, we examined the internalization and transport of DARP-36aa, a synthetic peptide derived from the N-terminal sequence of DARP, in rat C6 glioma cells. A colloidal gold DARP-36aa conjugate (DARP-36aa: AU) and biotinylated DARP 36aa were employed to visualize internalization and intracellular transport of DARP-36aa. Electron microscopy demonstrated that DARP-36aa: AU was rapidly incorporated into C6 glioma cells. Internalization via clathrin-coated pits and vesicles was clearly observed followed by transport and sorting of DARP-36aa: AU into multivesicular bodies, tubulo-vesicular endosomes, and lysosomes. Internalization of DARP-36aa: AU was also examined in primary mesencephalic cell cultures where a similar pattern of internalization and transport via clathrin coated pits and vesicles was observed. Fluorescence microscopy using a biotinylated DARP-36aa/avidin-rhodamine conjugate revealed that DARP-36aa is diffusely distributed on the plasmalemma prior to internalization at 4 degrees C. Following a 30-min incubation at 37 degrees C DARP-36aa was concentrated in the cytosol, particularly in areas surrounding cellular projections and the perikaryon. Immunocytochemical studies employing biotinylated DARP-36aa and an anti-clathrin heavy chain antibody demonstrated that DARP-36aa and clathrin colocalize during DARP-36aa internalization. We also observed a marked increase in tyrosine phosphorylation of a 45-kD protein in response to DARP-33a stimulation in C6 glioma cells. Genistein, a specific tyrosine kinase inhibitor, significantly inhibited DARP-36aa: AU internalization and transport in C6 glioma cells. These findings suggest that tyrosine kinase activity may result in DARP 36aa receptor-mediated endocytosis. PMID- 11867939 TI - Up-regulation of vasopressin and angiotensin II receptors in the thalamus and brainstem of inbred polydipsic mice. AB - Vasopressin (AVP), angiotensin II (Ang II) and oxytocin (OT) receptors were mapped in the brain of inbred polydipsic mice of the STR/N strain by quantitative in vitro autoradiography and receptor binding levels, compared with those found in control non-polydipsic mice of the ICR strain. A remarkable difference was evidenced in the thalamic paraventricular nucleus where AVP receptor binding was 7- to 10-fold higher in polydipsic mice than in control mice. Another disparity was observed in the hypothalamic paraventricular nucleus, which contained AVP binding sites in the control mice, but was unlabelled in the polydipsic animals. Ang II receptor binding was reduced in the hypothalamic paraventricular nucleus of the polydipsic mice, whereas it was abundant in the brainstem region, encompassing area postrema and the nucleus of the solitary tract. The distribution and amount of OT receptor binding were similar in the polydipsic and control mice. Strain-related differences of AVP and Ang II receptor binding were observed both in male and female animals. A sex-related difference was seen only for OT receptor binding in the hypothalamic ventromedial nucleus, where labelling was less intense in males than in females of both strains. Altogether, our results support the view that central AVP and Ang II systems are involved in the mechanisms responsible for polydipsia in STR/N mice. PMID- 11867941 TI - Elevated expression of galanin receptors in childhood neuroblastic tumors. AB - The neuropeptide galanin (GAL) has been shown to be present in certain brain tumors. In order to learn more about GAL and its receptors in human tumors of the peripheral nervous system, we investigated the expression of the GAL peptide and the GAL receptors in tumor tissue from childhood neuroblastic tumors. GAL peptide concentrations up to 674 +/- 166 fmol/mg of tissue were detected by radioimmunoassay, but no significant correlation with standard tumor markers or the prognosis of the 14 patients investigated was observed. Ligand binding experiments showed different levels of GAL binding in all 28 primary neuroblastomas and 7 ganglioneuromas investigated. All three human GAL receptor subtypes cloned to date could be detected, with the GALR1 receptor subtype being expressed most prominently. GAL binding did not significantly correlate with genetic markers such as unfavorable DNA ploidy, amplification of the oncogene MYCN and allelic loss of chromosome 1p. However, low galanin binding was significantly correlated with survival (p = 0.021) in this limited analysis of neuroblastic tumor samples. These results raise the possibility that the expression of GAL binding sites may play a role in neuroblastic tumor biology and behavior. PMID- 11867942 TI - Functional insulin-like growth factor-1/insulin-like growth factor-1 receptor mediated circuit in human and murine thymic epithelial cells. AB - Interactions between thymocytes and thymic epithelial cell (TEC) can be modulated by growth hormone via insulin-like growth factor-1 (IGF-1). In this study, we showed IGF-1 and IGF-1 receptor mRNA expression by human and murine TEC and thymocytes. Functionally, IGF-1 stimulates extracellular matrix production by human TEC. Moreover, pretreatment of murine TEC with IGF-1 increases their adhesion to thymocytes. Interestingly, we observed an increase in the frequency of CD4-CD8-CD90+ T cells which adhered to pretreated TEC, supporting the concept that IGF-1 may also act indirectly on intrathymic T cell differentiation and migration through the thymic epithelium. PMID- 11867940 TI - Increased response of plasma allopregnanolone to corticotropin-releasing hormone in obese patients. AB - Allopregnanolone is a neuroactive steroid present in the brain, but also measurable in systemic circulation. It exhibits anxiolytic and anticonvulsant effects and is able to produce hyperphagia. Since eating behavior disturbances and increased peripheral basal sympathetic activity have been reported in obese subjects, the present study investigated allopregnanolone and catecholamine (epinephrine and norepinephrine) responses to corticotropin-releasing hormone (CRH) in obese subjects. Blood was sampled from 39 obese (14 men and 25 women) and 57 normal-weight subjects (20 men and 37 women) and assayed for cortisol, allopregnanolone and catecholamines concentrations. In addition, 13 obese patients (5 men and 8 women) and 18 control subjects (9 men and 9 women) were submitted to a CRH test. Plasma allopregnanolone and norepinephrine levels were significantly higher in obese than in control subjects (p < 0.01), but plasma cortisol and epinephrine concentrations were comparable in both groups. No correlation was found in any group between plasma allopregnanolone and norepinephrine or epinephrine levels. Acute CRH administration significantly stimulated allopregnanolone secretion, with peak levels at 15 min in obese subjects, whereas maximal concentrations were reached after 60 min only in controls. In obese patients the allopregnanolone secretory incremental area was significantly higher than in controls (p < 0.02). CRH injection increased cortisol levels to a comparable extent in both groups. Plasma norepinephrine or epinephrine levels did were not significantly affected by CRH test in either group. In conclusion, obese subjects present higher allopregnanolone levels and a greater response to CRH than normal subjects. Since allopregnanolone has hyperphagic effects in rats, hypersecretion of the hormone in obese patients may represent one of the mechanisms underlying obesity. PMID- 11867943 TI - Hepatitis B virus and renal transplantation. PMID- 11867944 TI - Value and determinants of urine anion gap. PMID- 11867945 TI - Angiotensin II type 1 receptor antagonist, losartan, causes regression of left ventricular hypertrophy in end-stage renal disease. AB - Left ventricular hypertrophy (LVH) commonly occurs in patients with end-stage renal disease (ESRD) and is an independent risk factor for cardiovascular events. Angiotensin II type 1 receptor (AT1-R) antagonists may be able to reverse LVH independent to the hypotensive effect in the ESRD setting. Thirty chronically hemodialyzed uremic patients with hypertension were randomly assigned to receive the AT1-R antagonist losartan (n = 10), the angiotensin-converting enzyme (ACD) inhibitor enalapril (n = 10), or calcium antagonist amlodipine (n = 10). Left ventricular mass (LVM) index was measured by echocardiography before and 6 months after treatment. The baseline demographic and clinical characteristics did not differ between the three groups. The mean baseline LVM index also did not differ in the three groups. After 6 months of treatment, losartan treatment significantly reduced the LVM index (-24.7 +/- 3.2%) than amlodipine (-10.5 +/- 5.2%) or enalapril (-11.2 +/- 4.1%) therapy. All three groups had a similar decrease in the mean blood pressure with treatment. The plasma angiotensin II concentration increased 5-fold with losartan treatment. In contrast, the plasma angiotension II concentration did not change with enalapril and only increased 2 fold with amlodipine. Thus, the present study indicates that losartan more effectively regresses LVH in patients with ESRD than do enalapril and amlodipine despite a comparable depressor effect between the three drugs. PMID- 11867946 TI - Intravenous immunoglobulin G in the treatment of patients with chronic glomerulonephritis: clinical experience lasting 15 years. AB - In our study, we collected data on 116 patients with biopsy-proven idiopathic or lupus glomerulonephritis who were treated with high doses of intravenous immunoglobulin G (IVIG) (Veinoglobuline or Immunovenin-intact). In all patients a severe nephrotic syndrome (edema, proteinuria >6 g/24 h, serum albumin <22 g/24 h) had been observed. 34 patients had renal failure (serum creatinine up to 504 micromol/l) and 96 hypertension. 98 patients were previously for a long time treated with corticosteroids, immunosuppressors and anticoagulants without any effect. 18 patients had no therapy before IVIG. IVIG had been applied in a dose of 85 mg/kg/24 h 3 times every other day. Depending on the clinical improvement afterwards (in case of therapy resistance or relapse) these boli had been repeated in 84 patients after 1 month (and every 3 months for maintenance of remission) to 7 years. Proteinuria disappeared and full remission occurred in 36 patients. Partial remission was present in 48 patients. 32 patients went into end stage renal failure and/or died (15 of them of a nonrenal cause). In 13/34 patients with impaired renal function serum creatinine levels go back to normal after treatment. Our results suggested that IVIG therapy may be recommended in patients unresponsive to aggressive conventional treatment. PMID- 11867948 TI - Changes in Mac-1 and CD14 expression on monocytes and serum soluble CD14 level during push/pull hemodiafiltration. AB - BACKGROUND/AIM: Employment of treated dialysate as replacement fluid raises concerns about exposure of patients to pyrogenic substances. This study was undertaken to evaluate the safety of treated dialysate as the replacement fluid for push/pull hemodiafiltration. METHODS: In the present study, changes in the expressions of Mac-1 and CD14 on monocytes, which are upregulated by monocyte activation, were analyzed by flow cytometry, and the serum level of sCD14 which elevates by monocyte activation was measured by enzyme-linked immunosorbent assay (ELISA) during treatment in 7 patients on hemodialysis with regenerated cellulose (RC) membrane, polysulfone (PS) membranes and by push/pull hemodiafiltration (HDF) with PS membranes in a cross-over fashion. RESULTS: During hemodialysis with RC, hemodialysis with PS or push/pull hemodiafiltration with PS, both Mac-1 and CD14 expressions on monocytes significantly increased by passing through the artificial kidneys, and, accordingly, the respective values downstream of the artificial kidneys were significantly higher than the predialysis values, even when the lipopolysaccharide level in dialysate was not detectable by Limulus assay. There was no significant variation in serum sCD14 levels during any of the hemodialysis with RC, hemodialysis with PS or push/pull hemodiafiltration. However, during hemodialysis with PS or push/pull hemodiafiltration with PS, changes in Mac-1 and CD14 expression on monocytes were significantly smaller than those during hemodialysis with RC. CONCLUSION: Monocytes are activated to a greater extent during hemodialysis with RC membranes than during push/pull HDF with PS membranes. We consider that push/pull HDF may be safer than hemodialysis with RC membrane and that it is as safe as hemodialysis with the PS membrane in terms of monocyte activation, when pyrogen-free dialysate is employed. PMID- 11867947 TI - The renal glomerulus and vasculature in 'aggregation' chimeric mice. AB - In glomerular development, the glomerular epithelium is derived from the lower loop of the S-shaped renal vesicle. However, it is unclear whether the capillary endothelium is derived directly by vasculogenesis (e.g. differentiated directly from local metanephric mesenchyme) or whether they are derived by angiogenesis (i.e. derived from pre-existing vasculature in the metanephros). This question has been addressed in other laboratories using surgically created chimeric kidney model systems. In the present study, chimeric kidneys were developed by aggregating the cells from 4- to 8-cell embryos from Mus musculus with ones from Mus caroli and implanting the aggregated embryos into pseudopregnant hosts [Goldowitz D: Neuron 1989;3:705-713]. Species specific DNA clones were used in conjunction with in situ hybridization to identify the species origin of cells. Interspecies aggregate chimeras had varying proportions of renal cells derived from Mus caroli and Mus musculus; however, regions were identified in which the renal tubular and Bowman's capsule or parietal epithelia were from one species while vessel endothelium and cells in the interstitium were from the other species. In those regions, glomeruli always contained an admixture of cells from both species however; many of the glomerular endothelial cells appear to be from the same species as the vessel endothelium and interstitial cells. These findings support the hypothesis that angiogenesis may contribute cells that help form the glomerular capillary endothelium. Most intrarenal arteries contained cells from both species. However a few vessels were found in which the endothelium was derived from one species while the smooth muscle cells were from the other species. This finding suggests that intrarenal arterial development has two cells of origin: the endothelial tube develops and is surrounded by mesenchymal cells that form the tunica media. The aggregation chimeric mouse kidney may become a useful model system for studying in situ aspects of the complex processes involved in kidney development. PMID- 11867949 TI - Insulin-like growth factor system components in relation to erythropoietin therapy and bone metabolism in dialyzed patients and kidney transplant recipients. AB - Insulin-like growth factor (IGF) system components appear to be the most important regulators of bone cell function. On the other hand, IGF-1 is shown to be an important regulator for erythropoiesis. The aim of the study was to examine the relationships between IGF system, requirements of erythropoietin, endogenous erythropoietin levels, bone metabolism assessed by biochemical markers, markers of nutrition such as cholesterol and albumin in recombinant human erythropoietin (rHuEPO)-treated patients maintained on chronic hemodialyses or peritoneal dialyses as well as in kidney transplant recipients. The studies were performed on 79 chronically hemodialyzed patients; 28 of them did not receive rHuEPO, 51 subjects received rHuEPO, 34 patients on continuous ambulatory peritoneal dialysis (CAPD), 16 of them did not receive rHuEPO, 18 were given rHuEPO and 46 kidney allograft recipients. Endogenous erythropoietin concentration, bone specific alkaline phosphatase and serum CrossLaps were assayed by ELISA. Intact PTH, osteocalcin, 1,25-(OH)(2) D(3), 25-OH D(3), IGF-1, procollagen type I carboxy-terminal extension peptide (PICP) and procollagen type I cross-linked carboxy-terminal telopeptide (ICTP) were studied by RIA, whereas IGFBP-1 and IGFBP-3 concentrations were assayed by IRMA. We found a significantly higher IGF 1 and IGFBP-3 in rHuEPO-treated HD patients when compared to CAPD subjects given rHuEPO as well as to hemodialysis (HD) patients not treated with rHuEPO. IGF-1 was significantly higher in kidney transplant recipients when compared to dialyzed patients without rHuEPO therapy. IGFBP-1 was similar in all groups of patients (including kidney transplant recipients) studied. In CAPD patients not given rHuEPO concentrations of ICTP and PICP were significantly lower when compared to rHuEPO-treated CAPD subjects and HD patients not receiving rHuEPO therapy. Serum CrossLaps in CAPD patients treated with rHuEPO were significantly higher when compared to CAPD subjects without rHuEPO treatment and to kidney transplant recipients. In rHuEPO-treated CAPD subjects IGF-1 and IGFBP-1 correlated positively with serum CrossLaps (r = 0.61, p < 0.05 and r = 0.64, p < 0.05, respectively), whereas in hemodialyzed patients without rHuEPO a significant negative correlation between IGFBP-3 and serum CrossLaps was found (r = --0.69, p < 0.001) as well as between IGFBP-3 and aluminium (r = 0.51, p < 0.05), IGF-1 and ICTP (r = --0.43, p < 0.05). In conclusion, our data indicate a probable functional relationship between IGF system components, erythropoietin treatment in dialyzed patients and bone metabolism in renal replacement therapy in a form of hemodialyses, peritoneal dialyses and kidney transplantation. Dialyzed patients exhibit more pronounced renal osteodystrophy than kidney allograft recipients. IGF system components are influenced by erythropoietin therapy, but are not related to serum erythropoietin levels and rHuEPO requirements. PMID- 11867950 TI - Comorbidity and mortality in peritoneal dialysis: a comparative study of type 1 and 2 diabetes versus nondiabetic patients. Peritoneal dialysis and diabetes. AB - We conducted a retrospective study with 750 peritoneal dialysis (PD) patients in a Spanish multicenter registry between 1993 and 1999 to analyze comorbidity and mortality in type 1 diabetes (T1D), type 2 diabetes (T2D) and nondiabetic (ND) patients. 163 patients (21.7%) were diabetic--96 T1D (58.8%) and 67 T2D (42.2%)- while 587 were not (78.3%). Different comorbidity factors such as the presence of cardiovascular disease, age over 70 and dyslipidemia at the start of PD were analyzed as well as the incidence of peritonitis, the peritonitis-free interval, need for hospitalization, mortality rate, early mortality rate, survival curves (log rank) and the impact factor (Cox) on mortality for the different variables. The comorbidity index (number of comorbidity factors when starting the treatment) and the peritonitis incidence were higher for T2D. Hospitalization rates were similar, but mortality rates were higher for T2D and early mortality rates (death during the 1st year of treatment) were higher for T1D. The actuarial survival curves showed a higher mortality for T2D with no differences between ND and T1D after adjustment for age. The mortality odds ratio was 1.78 for T2D and 1.13 for T1D, differences which were not significant after adding age over 70 and cardiovascular disease to the variables analyzed. Our results show that associated comorbidity is the most important difference between ND, T1D and T2D. While cardiovascular comorbidity is responsible for the higher percentage of early mortality found in T1D when compared to ND, both age and cardiovascular disease are responsible for the higher comorbidity and mortality found in T2D. PMID- 11867951 TI - Combination therapy with benazepril and oral adsorbent ameliorates progressive renal fibrosis in uremic rats. AB - BACKGROUND/AIMS: The administration of an angiotensin-converting enzyme (ACE) inhibitor or an oral adsorbent, AST-120 (Kremezin), prevents the progression of renal failure. This study was designed to determine the additional effects of AST 120 combined with an ACE inhibitor, benazepril, on the progression of renal fibrosis in uremic rats. METHODS: 5/6-nephrectomized uremic rats were divided into control uremic rats (CRF group), benazepril-treated uremic rats (CRF+B group) and uremic rats receiving benazepril and AST-120 (CRF+BK group). After 14 weeks of treatment renal function and pathological changes were investigated. RESULTS: The progression of renal dysfunction was delayed in both the CRF+B and CRF+BK groups as compared with the CRF group. In the CRF+BK group, the level of serum and urinary indoxyl sulfate and the tubular accumulation of indoxyl sulfate decreased. Both the CRF+B and CRF+BK groups showed lower glomerular sclerosis indices than the CRF group. In the CRF+BK group, but not the CRF+B group, the interstitial fibrosis area and the expression of transforming growth factor (TGF) beta1 and tissue inhibitor of metalloproteinases (TIMP) 1 were decreased as compared with the CRF group. Furthermore, the CRF+BK group showed a smaller interstitial fibrosis area and a lower renal osteopontin expression than the CRF+B group. CONCLUSION: Combination therapy of benazepril and AST-120 is more effective than benazepril alone in retarding the progression of interstitial fibrosis by reducing the expression of TGF-beta 1, TIMP-1 and osteopontin. PMID- 11867952 TI - Dietary quercetin augments activator protein-1 and does not reduce nuclear factor kappa B in the renal cortex of rats with established chronic glomerular disease. AB - BACKGROUND/AIMS: Quercetin, a naturally occurring dietary flavonoid, is a potent in vitro inhibitor of the transcription factors, nuclear factor (NF)-kappa B and activator protein (AP)-1. Here we investigated the efficacy of quercetin to suppress renal cortical NF-kappa B/AP-1 activation and tubulointerstitial injury, in vivo, in a nephrotic rat model of chronic glomerular disease. METHODS: Adult male Wistar rats with (n = 27) or without (n = 15) Adriamycin nephropathy (AN) were stratified into six groups (according to proteinuria and endogenous creatinine clearance, measured on day 10) and pair-fed a semipurified diet supplemented with or without quercetin (0.5, 2%) from day 14 to 36. RESULTS: Quercetin-fed rats had minimal weight gain during the study (p < 0.05). In AN, proteinuria, the decline in endogenous creatinine clearance, hypercholesterolaemia and cortical tubulointerstitial injury were not reduced by quercetin. Quercetin had no effect on renal cortical NF-kappa B activation, but unexpectedly exacerbated the induction of AP-1 in AN (by 67%, p < 0.05). In normal rats, quercetin increased proteinuria (by 75%, p < 0.05), renal cortical AP-1 activation (by 198%, p = 0.06), and malondialdehyde production (by 110%, p < 0.05). CONCLUSION: These data suggest that supplementation of the diet with quercetin is not an effective therapeutic strategy alone to reduce NF-kappa B/ AP 1 activation in chronic glomerular diseases, and paradoxically could be associated with adverse renal effects. PMID- 11867953 TI - Experimental nephropathy induced by Haemophilus parainfluenzae antigens. AB - BACKGROUND: We have demonstrated that outer membrane antigens of Haemophilus parainfluenzae (OMHP) are potentially involved in IgA nephropathy (IgAN). In this study, we established an experimental model of IgAN using OMHP antigens and investigated the nephritogenicity of OMHP antigens. METHODS: One hundred and twenty C3H/HeN mice were administered OMHP antigens orally (PO group) or intraperitoneally (IP group). Mice were sacrificed at 10, 20, 30, 40, and 50 weeks of age to examine sequential glomerular changes and to measure levels of IgG, IgA, and IgM antibody against OMHP by ELISA. RESULTS: Glomerular deposition of IgA and increases in the amount of mesangial matrix were observed in the PO group and the IP group from 40 and 30 weeks of age, respectively. Mice in both groups showed glomerular deposition of OMHP antigens from 30 or 40 weeks of age. Levels of IgA antibodies against OMHP were significantly increased in the PO and IP groups compared with controls. There was a significant correlation between mesangial proliferation and glomerular deposition of IgA. CONCLUSIONS: Administration of OMHP antigens to mice may induce glomerular deposition of IgA and mesangial proliferation, resembling the changes seen in IgAN, with increases in IgA antibodies against OMHP antigens. This is the first use of OMHP antigens to establish an active model of IgAN. PMID- 11867954 TI - Tryptophan metabolism via the kynurenine pathway in experimental chronic renal failure. AB - BACKGROUND: Kidneys are involved in tryptophan (TRP) metabolism in two ways. They eliminate TRP derivatives on the one hand, and they produce several enzymes taking part in TRP metabolism mainly via the kynurenine pathway on the other. The aim of the present study was to examine the time-course of changes in the peripheral kynurenine products degradation during experimental chronic renal failure in rats. METHODS: Tryptophan, kynurenine, 3-hydroxykynurenine, kynurenic acid, xanthurenic acid, anthranilic acid and quinolinic acid were determined in plasma using high-performance liquid chromatography technique with UV, fluorescence and electrochemical detection. RESULTS: A decreased TRP level and significant increase in kynurenine pathway metabolite concentrations in plasma of uremic rats were found. CONCLUSIONS: Substantial disturbances in the peripheral kynurenic pathway were observed in experimental chronic renal failure. They may contribute to several symptoms of uremia. PMID- 11867955 TI - Remission of nephrotic syndrome due to AA amyloidosis and initiation of glomerular repair after surgical resection of localized Castleman's disease. AB - To shed further light on the eventual destiny of amyloid kidney deposits after interruption of amylogenic stimulus, we report a case of a 47-year-old woman with nephrotic syndrome due to renal amyloidosis, complicating abdominal Castleman's disease. After 5 courses of therapy with melphalan and prednisolone which failed to improve the nephrotic syndrome or her general clinical condition, and 1 year after the diagnosis of renal amyloidosis, surgical excision of the abdominal mass was performed. Whereas her clinical symptoms and other laboratory findings rapidly improved, the proteinuria took 18 months to disappear. A second renal biopsy, performed 30 months after surgical resection, showed persistence of the amyloid deposits in the same extent. However, electron microscopy revealed subtle reparative phenomena at the epithelial site of the basement membrane. We conclude that proteinuria associated with amyloidosis does not only depend on structural damage and that the new synthesized segment of basement membrane observed by us probably represents a mechanism of repair and the start of a long healing process. PMID- 11867956 TI - Elevated plasma vascular endothelial growth factor levels in non-diabetic predialysis uraemia. AB - Chronic renal impairment is associated with increased vascular morbidity and mortality. The pathogenesis of this aggressive vascular pathology is unknown but believed to be multi-factorial in origin. There is accumulating in vivo and in vitro evidence to suggest that the vascular endothelium is dysfunctional in uraemia, including the propensity of endothelial cells to produce VEGF in response to acidosis. There is also preliminary data to suggest abnormally increased endothelial permeability in uraemia. To investigate the potential abnormal circulating levels of VEGF in uraemia, EDTA plasma samples were collected from 20 non-diabetic predialysis patients and matched controls. Free plasma VEGF levels were detected using a commercially available ELISA kit. There were significantly higher plasma levels of VEGF predialysis group (median 351 pg/ml, range 70-636 pg/ml) compared to matched controls (median 125.5 pg/ml, range 22-450 pg/ml), p < 0.002. In conclusion, free plasma VEGF levels are high in chronic renal impairment. We hypothesise that this potent growth and permeability factor may contribute to the endothelial dysfunction of uraemia. PMID- 11867957 TI - Prevalence of Cryptosporidium infection in hemodialysis patients. AB - The prevalence of Cryptosporidium infection in patients who are on chronic hemodialysis due to end-stage renal failure is investigated and compared with the incidence in the healthy population. Stool specimens of 74 adult hemodialysis patients treated on an outpatient basis and 50 healthy individuals are examined for Cryptosporidium oocysts by using the modified acid-fast method. While 20.27% (15/74) of patients in the dialysis group had Cryptosporidium oocysts in their stools, none (0/50) of the controls had such an infection (p < 0.001). Ten of 15 patients who had Cryptosporidium oocysts in their stool specimens were asymptomatic and the remaining 5 had diarrhea. Four of these 5 diarrheic patients had Cryptosporidium oocysts in their stools and 1 had both Cryptosporidium oocysts and Giardia trophozoites. Since hemodialysis patients with chronic renal failure are candidates for renal transplantation, general preventive measures against infection must be taken and these patients must be screened for their carrier state before transplantation. PMID- 11867958 TI - Beta(2)-microglobulin clearance decreases with Renalin reuse. AB - The HEMO study revealed that beta(2)-microglobulin clearance decreases over time with Renalin reuse in the high-flux group. It was suggested that the reuse of polysulfone or cellulose triacetate high-flux dialyzers with Renalin (without bleach) results in degradation of the high-flux capacity. At our haemodialysis unit (Vila Real, Portugal) we reused dialyzers until January 2000 (limited to 10 reuses), with an automatic machine Renatron (Renal Systems, Minntech. All of our 31 patients who started with postdilution haemodiafiltration on-line (HDFol) were always dialyzed with F-80 polysulfone (Fresenius). The reposition rate was 10 litres/session until 1998 and 20 litres/session thereafter. Reuse techniques were abandoned in our country in January 2000 following an EEC directive. Thereafter, we have decided to maintain HDFol with the same dialyzers without reuse. The mean beta(2)-microglobulin predialysis values did not decrease over time until reuse was terminated (1995 with low-flux haemodialysis: 25.4 +/- 6.4 microg/l; 1997: 24.7 +/- 6.6 microg/l; 1998: 29.2 +/- 8.9 microg/l; 1999: 33.7 +/- 4.7 microg/l) whereas beta(2)-microglobulin clearances were reasonable with HDFol (1998: 56.4 +/- 25.9 ml/min; 1999: 47.9 +/- 16.4 ml/min). After stopping reuse we have noticed that predialysis beta(2)-microglobulin values decreased (2000: 23.0 +/- 3.9 microg/l) in accordance with beta(2)-microglobulin clearance duplication (2000: 84.1 +/- 25.0 ml/min; p < 0.01). It is our opinion that the reuse of polysulfone dialyzers with Renalin should be abandoned in the field of high-flux haemodialysis. It causes deterioration in the beta(2)-microglobulin clearance and probably interferes with the high-flux haemodialysis benefits, namely the reduction of dialysis-related amyloidosis. PMID- 11867959 TI - Reflex anuria from unilateral ureteral obstruction. AB - Renal function is usually normal or only marginally affected in patients with unilateral ureteral obstruction due to the vicarious function of the contralateral kidney. Few reports exist in which unilateral renal obstruction is associated with anuria (reflex anuria, RA) and acute renal failure. We report the clinical case of a female patient who was referred to the emergency department due to anuria of 72 h duration and acute renal failure (serum creatinine 9 mg/dl) associated with several episodes of violent right flank pain with hematuria following extracorporeal shock wave lithotripsy (ESWL). A few weeks before ESWL, urography showed a 2-cm stone located in the right pelvis whilst the left kidney was functionally normal. On admission, renal ultrasound documented a normal left kidney, whilst the right pelvis was hydronephrotic and there were two indwelling stones at the right pyeloureteral junction. After the patient passed a urinary stone, diuresis restarted and acute renal failure was resolved. Thereafter, urography confirmed that the left kidney, the left ureter and bladder were functionally and morphologically normal. RA with acute renal failure has been so scarcely documented that it is considered to be legend by many clinicians. Major textbooks do not discuss RA with acute renal failure. Vascular or ureteral spasm related in part to a peculiar hyperexcitability of the autonomic nervous system may explain RA. We suggest that nephrologists should always consider RA when evaluating acute renal failure. On the other hand, RA might be relatively common and we cannot rule out that only the most severe and/or better-documented cases have been reported in the medical literature. PMID- 11867960 TI - Stomach phytobezoars in two uremic anorexic patients. AB - Conglomerates of food and mucus or phytobezoars composed of vegetable matter are sometimes found in the stomach in the general population. Reports of phytobezoars in uremic patients are, however, scarce. Here we describe 2 uremic patients in which esophagogastroduodenoscopy was performed due to dyspepsia associated with weight loss and in which stomach phytobezoars were discovered. Theoretically, uremic patients should be at risk for producing bezoars. In fact, these patients frequently present predisposing conditions such as autonomic neuropathy, diabetes mellitus and delayed gastric emptying. Gastric bezoars cause anorexia. Anorexia is a frequent symptom in dialysis patients and is associated with malnutrition. In these patients, malnutrition is strongly associated with mortality and is quite difficult to reverse. Similarly, phytobezoars cause chronic anorexia. We suggest that clinicians working in dialysis units should consider the possibility of a gastric bezoar when evaluating anorexic uremic patients. PMID- 11867961 TI - High level of negative emotions in hemodialysis patients suggests the need for therapy. PMID- 11867963 TI - High-frequency ultrasound in the evaluation of pharmacological effects on the skin. AB - In 1987, the introduction of two 20-MHz B- (brightness-) scan ultrasound systems with axial and lateral resolution of 200 microm and 75-80 microm, respectively, rendered two-dimensional sonographic imaging in dermatology possible. Since then cutaneous sonography has increasingly been used to investigate drug effects on human skin. Both unwanted and wanted effects have been studied, including skin thinning due to repeated glucocorticoid application and reduction of skin thickening in hyperplastic disorders such as psoriasis vulgaris due to various treatment approaches. The resolution of 20-MHz ultrasound is sufficient for studying pathological changes and pharmacological effects in the dermis and the subcutaneous fat; for assessing the epidermis, frequencies between 50 and 100 MHz may prove to be more effective. PMID- 11867962 TI - Novel trial for the treatment of steroid dependency in minimal change disease: combined therapy of camostat mesilate and glycyrrhizin. PMID- 11867964 TI - Different skin thinning potential of equipotent medium-strength glucocorticoids. AB - In this study, we investigated the effect of prednicarbate, mometasone furoate and betamethasone 17-valerate on total skin thickness over a treatment period of 6 weeks. The study was conducted as a double-blind, placebo-controlled randomized clinical trial with a confirmatory approach. The influence of these drugs on healthy human skin under non-occlusive conditions was assessed by measuring total skin thickness and epidermal thickness using 20 and 50 MHz sonography, respectively. Epidermal surface structure was evaluated using profilometry. Visual assessment addressed signs of atrophy and formation of telangiectasia. The reduction of total skin thickness induced by prednicarbate was clearly less than that caused by betamethasone 17-valerate and mometasone furoate. Prednicarbate led to a higher degree of skin thinning than vehicle. For technical reasons, epidermal thickness could not be reliably evaluated with 50 MHz sonography. Profilometry did not demonstrate any differences between treatments. Visible signs of atrophy or telangiectasia were detected in two subjects each upon betamethasone 17-valerate and mometasone furoate, but not upon prednicarbate or its vehicle. Prednicarbate is a topical glucocorticoid with an improved benefit/risk ratio, as it causes less skin atrophy than the equipotent betamethasone 17-valerate. PMID- 11867965 TI - Flow cytometric assessment of clearance and relapse characteristics in psoriasis vulgaris after treatment with weekly clobetasol lotion under hydrocolloid occlusion versus twice-daily clobetasol ointment. AB - Clearance and relapse characteristics of clobetasol lotion under hydrocolloid occlusion once weekly versus clobetasol ointment twice daily were assessed in a comparative flow cytometric study. Quantitative analysis of markers for epidermal proliferation, differentiation and inflammation was performed on epidermal single cell suspensions prepared from 3-mm punch biopsies taken from 15 patients with psoriasis vulgaris before therapy, at clearance and 6 weeks after clearance. After treatment both therapy regimens resulted in substantial changes of all flow cytometric parameters, but clearance was induced earlier in the corticosteroid under hydrocolloid occlusion-treated group. With respect to the relapse phase no difference was observed between both treatments. Although it is remotely possible that the outcome in the treatment of more extensive psoriatic lesions might be different, the present study suggests that the robust clinical efficacy of the treatment with a topical corticosteroid under hydrocolloid occlusion is not associated with a rebound phenomenon. PMID- 11867966 TI - Modulation of atopy patch test reactions by topical treatment of human skin with a fatty acid-rich emollient. AB - Measures directed at improving the skin barrier function are thought to be effective in preventing reexacerbation of atopic dermatitis, but direct proof of a prophylactic effect of emollients has been elusive. In the present study, the atopy patch test has been employed as a model for the initiation phase of atopic dermatitis in order to assess whether pretreatment of non-lesional skin with a fatty acid-rich emollient (Eucerin Omega Creme) has a prophylactic effect in patients with atopic dermatitis. Pretreatment of test sites with Eucerin Omega Creme either prevented or diminished the development of eczema, as compared with untreated control test sites in the same patients (n = 38). These studies indicate that the use of fatty acid-rich emollients prevents the development of atopic eczema. They also demonstrate that the atopy patch test can be used to assess the capacity of a given regimen to exert prophylactic effects in this disease. PMID- 11867968 TI - Lymecycline and minocycline in inflammatory acne: a randomized, double-blind intent-to-treat study on clinical and in vivo antibacterial efficacy. AB - BACKGROUND: Some antibiotics represent a mainstay in acne treatment. However, studies comparing their efficacies are rare. AIM: To evaluate the clinical and in vivo antibacterial effect of lymecycline and minocycline at different dosages. METHOD: Eighty-six patients with moderate to severe acne were enrolled in a randomized, double-blind, intent-to-treat study comparing in three parallel groups the effect of (1) lymecycline 300 mg daily for 12 weeks, (2) minocycline 50 mg daily for 12 weeks and (3) minocycline 100 mg daily for 4 weeks followed by 50 mg daily for 8 weeks. Evaluations were made at the screening visit and at five on-treatment visits. They consisted of clinical counts of acne lesions and evaluations of bacterial viability using dual flow cytometry performed on microorganisms collected from sebaceous infundibula by cyanoacrylate strippings. RESULTS: Patients receiving minocycline 100/50 mg had the best clinical outcome, particularly in the reduction of the number of papules. By the end of the trial, the microbial response to minocycline 100/ 50 mg was also superior to either of the other two treatments. There were less live and more dead bacteria. CONCLUSION: In this trial, minocycline 100/50 mg was superior for the treatment of inflammatory acne when compared to lymecycline 300 mg and minocycline 50 mg. PMID- 11867967 TI - Comparison of analgesic and anti-inflammatory activity of meloxicam gel with diclofenac and piroxicam gels in animal models: pharmacokinetic parameters after topical application. AB - Meloxicam, a non-steroidal anti-inflammatory drug, is a preferential inhibitor of cyclooxygenase-2 and has demonstrated potent analgesic and anti-inflammatory activity after oral administration. The present work was carried out to elucidate the anti-inflammatory and analgesic activity of a newer topical gel formulation of meloxicam (1% w/w gel) and compare it with 0.5% w/w piroxicam and 1% w/w diclofenac gels in experimental animal models. The study was also extended to determine the pharmacokinetic profile of a newer formulation of meloxicam gel after topical application on depilated skin of rats. The anti-inflammatory activities of meloxicam, piroxicam and diclofenac gels were compared using carrageenan-induced acute paw oedema and complete Freund's adjuvant-induced chronic paw oedema in rats. Meloxicam gel showed increased protection against inflammation as compared to piroxicam and diclofenac gels. Acetic acid-induced writhing and formalin-induced phase I and phase II pain models were used to compare their analgesic activity. Meloxicam gel showed significant protection in formalin-induced phase II pain whereas its analgesic activity was less as compared to diclofenac and piroxicam gels in writhing test and formalin-induced phase I pain. The pharmacokinetic studies showed peak plasma drug concentration (C(max)) of 48.48 +/- 6.57 microg/ml at 2 h (T(max)) after topical application of 500 mg of meloxicam gel formulation. The area under the curve as calculated from 0 to 6 h was found to be 114.18 +/- 4.23 and 194.13 +/- 3.78 microg x h/ml for 0 to infinity. The results indicate that topical preparation of meloxicam could be an effective alternative to diclofenac and piroxicam gels in inflammatory conditions and its associated pain with the possibility of less systemic side effects. PMID- 11867970 TI - Anti-inflammatory efficacy of topical preparations with 10% hamamelis distillate in a UV erythema test. AB - In 40 volunteers the efficacy of three lotions with 10% hamamelis distillates from different suppliers, two vehicles, dimethindene maleate 0.1% gel, hydrocortisone 1% cream and hydrocortisone 0.25% lotion were investigated in a modified UV erythema test with three UV dosages (1.2, 1.4 and 1.7 MED). The test preparations were applied occlusively over a 48-hour period following irradiation. Chromametric measurement of redness and visual assessment were performed 24, 48 and 72 h after induction of erythema. The hydrocortisone formulations were most effective in erythema suppression. An anti- inflammatory effect was noted for all three hamamelis lotions as well as for the vehicles. A significantly greater suppression of erythema than seen with the vehicles was noted for one of the hamamelis lotions at 1.4 MED. The efficacy of the antihistamine dimethindene maleate did not surpass the hamamelis lotions or the vehicles. Even though the differences between the hamamelis lotions were slight, it was possible to make an objective selection of the best hamamelis distillate for aftersun purposes. PMID- 11867969 TI - Modulating effects of oatmeal extracts in the sodium lauryl sulfate skin irritancy model. AB - The aim of the present study was to assess the anti-inflammatory activity of two topically applied oatmeal extracts, i.e. Avena sativa and Avena Rhealba, using the sodium lauryl sulfate (SLS) irritation model. At baseline, test areas on the volar surface of the upper arms of 12 healthy individuals were pretreated with the two extracts and their vehicle (petrolatum ointment) under occlusion for 2 h, and one site was left untreated. Then a patch with a 1% SLS solution was applied to the test sites for 24 h. Irritation was determined at each period by measuring by chromametry and laser-Doppler. In a dose-ranging study with the Avena Rhealba extract alone, the 20 and 30% concentrations exerted a slight inhibition of the a* parameter increase and a marked reduction of the blood flow increase (p < 0.05, compared to vehicle). Then, the effects of the two extracts at the concentration of 20% were compared. All extracts displayed a statistically significant counteracting effect on both parameters (p < 0.05), but no statistically significant difference between treatment groups could be demonstrated. In conclusion, this study demonstrates the preventive effects of oatmeal extracts on skin irritation in the SLS model. PMID- 11867971 TI - Comparative development of SIRC rabbit corneal cells grown on polycarbonate- and polyester-based filters. AB - Statens Seruminstitut Rabbit Cornea (SIRC) cells were grown on sterile polycarbonate- and polyester-based filter inserts to evaluate the feasibility of substituting the polyester-based filter inserts for use in a previously developed permeability model. The two filter types were compared for differences in cellular growth rate and morphological changes following days 3, 7 and 10 in culture. There were no significant differences in the number of cell layers formed on any day examined. The numbers of cell layers formed with the polycarbonate-based filter inserts were 1.75 +/- 0.09, 2.53 +/- 0.14 and 2.93 +/- 0.16, for days 3, 7 and 10, respectively. The numbers of cell layers formed with the polyester-based filter inserts were 1.79 +/- 0.07, 2.23 +/- 0.11 and 2.90 +/- 0.13, for days 3, 7 and 10, respectively. The SIRC cells had a similar comparative morphology on each of the above days. Active cell growth was seen on each day with signs of maturation evident by day 7 in culture. The polyester based filter inserts can be substituted in the previously established permeability assay without alteration of morphology or rate of growth, thus allowing confirmation of cell confluency prior to use in the permeability assay. It will also allow for photographic documentation of cell injury prior to recovery studies, without necessitating the preparation of extra samples for fixation at the time of initial injury. Future studies will be required to determine if there will be any alteration in the rate of permeability with a previously tested standard, before adoption of the new filter. PMID- 11867972 TI - Treatment patterns of isolated benign headache in US emergency departments. AB - STUDY OBJECTIVE: I sought to describe and analyze the treatment of a large representative sample of adult US emergency department patients with isolated primary headache. METHODS: Information on adult patients with an isolated diagnosis of migraine headache or unspecified headache was extracted from the 100.4 million ED visits represented by the 1998 National Hospital Ambulatory Medical Care Survey. Demographic and clinical information are presented with descriptive statistics. The treatment of migraine headache was assessed in light of Canadian and US practice parameters. RESULTS: The migraine headache and unspecified headache cohorts included 811,419 and 604,977 participants, respectively. The majority of patients were young, white, and female. Patients received a mean of 1.8 medications from a pharmacopoeia of 36 drugs. Most patients (84.8%) given a diagnosis of migraine headache received a parenteral agent. The most commonly used medications were meperidine (30.0%), ketorolac (21.4%), and prochlorperazine (16.7%). Adjunct antiemetics were commonly administered with parenteral opioids (89.8%). Promethazine and hydroxyzine, antiemetics without anti-headache effects, were used 6 times more commonly as adjuncts than the dopamine antagonists that have established anti-headache effects (ie, prochlorperazine, metoclopramide, droperidol; 78.0% versus 11.8%). The US and Canadian recommendations for the use of nonopioid abortive medications (dopamine-antagonist antiemetics, dihydroergotamine, and 5-hydroxytrypamine(1) [5 HT(1)] receptor agonists) are supported by strong evidence. However, parenterally treated patients with migraines received opioids as their only anti-headache medication more commonly than they received any of the aforementioned non-opioids in their regimen (45.7% versus 26.0%). Of all the opioid recipients, most (77%) did not receive any nonopioid abortive headache medication. Meperidine was the most commonly administered opioid (70%). CONCLUSION: Polypharmacy and a broad pharmacopoeia characterize the US ED treatment of isolated benign headache. Opioid use, particularly meperidine, exceeds that of recommended nonopioid abortive migraine medications. PMID- 11867973 TI - Effect of vital signs on triage decisions. AB - STUDY OBJECTIVE: We sought to determine whether knowledge of vital signs changes nurse triage designations (TDs). We also sought to determine whether patient age and ability to communicate modify the effect of vital signs on triage decisions. METHODS: We performed a prospective observational study, in 24 emergency departments, of nurse-assigned TDs of all ED patients undergoing triage. Nurses performed their typical triage routine, except that they chose 1 of 5 hypothetical TDs (call 911, ED <2 hours, physician's office 2 to 8 hours, physician's office 8 to 24 hours, or home care) before and after measurement of vital signs. The main outcome measure was the change of TD after knowledge of a patient's vital signs, with stratification on the basis of patient age and communication barriers. The secondary outcome was the final ED disposition. RESULTS: Six hundred twenty-five experienced triage nurses at 24 different EDs collected data on 14,285 patients. TDs were downgraded (decreased in urgency) in 2.4% of patients, and 5.5% were upgraded (increased in urgency) after vital signs were known. Changes were more likely to occur in the young (< or = 2 years old; 11.4%) and the elderly (> or = 75 years old; 9.9%) than in those 3 to 74 years of age (7.5%). When nurses reported a communication barrier, a change in post-vital signs TD was also more common (11.2% versus 7.7%). The post-vital signs TD better predicted patient ED disposition. CONCLUSION: In this sample, 92.1% of the nurses' TDs were not affected by the knowledge of patient vital signs. For the other 7.9%, including many patients from vulnerable populations, the vital signs changed the nurses' assessments of the patients' triage designation. Methods of triage that do not determine vital signs may not adequately reflect the urgency of the patient's presentation. PMID- 11867974 TI - The reliability of vital sign measurements. AB - STUDY OBJECTIVE: Vital sign measurements, specifically heart rate, respiratory rate, and blood pressure, play a fundamental role in many medical evaluations, yet little is known about the reliability of noninvasive vital sign measurements. We sought to determine whether trained observers can reproducibly assess vital signs in the clinical setting. METHODS: Two trained observers independently measured vital signs on 140 patients presenting to an urban emergency department with acute medical complaints. Heart rate and respiratory rate were each measured by auscultation of heart and breath sounds for 1 minute. Systolic and diastolic blood pressures were determined by auscultating Korotkoff sounds while viewing pressure measurements from a standard cuff and mercury manometer. The mean value of each vital sign and Bland-Altman statistics (mean difference between observers [MDO], expected range of agreement [ERA]) were used to provide absolute and relative indices of reliability. RESULTS: The observers found a mean heart rate of 78.5 beats/min, with an MDO of 0.02 beats/min (0.03%), and an ERA of +/- 10.6 beats/min (+/- 13.5%). Respiratory rate exhibited a mean of 17.5 breaths/min, an MDO of 0.04 breaths/min (0.2%), and an ERA of +/- 6.2 breaths/min (+/- 35.5%). The mean systolic blood pressure of 127.1 mm Hg was associated with an MDO of 1.3 mm Hg (1.0%), and an ERA of +/- 24.2 mm Hg (+/- 19.0%). Diastolic blood pressure exhibited a mean of 77.4 mm Hg, an MDO of 0.3 mm Hg (0.4%) with an ERA of +/- 19.9 mm Hg (+/- 25.7%). CONCLUSION: The reproducibility of vital sign measurements may be limited by significant interobserver variability. Clinicians should recognize this inherent variability and interpret vital signs with caution. PMID- 11867975 TI - Older adults in the emergency department: a systematic review of patterns of use, adverse outcomes, and effectiveness of interventions. AB - STUDY OBJECTIVE: We sought to synthesize the literature on patterns of use of emergency services among older adults, risk factors associated with adverse health outcomes, and effectiveness of intervention strategies targeting this population. METHODS: Relevant articles were identified by means of an English language search of MEDLINE, HealthSTAR, CINAHL, Current Contents, and Cochrane Library databases from January 1985 to January 2001. This search was supplemented with literature from reference sections of the retrieved publications. A qualitative approach was used to synthesize the literature. RESULTS: Compared with younger persons, older adults use emergency services at a higher rate, their visits have a greater level of urgency, they have longer stays in the emergency department, they are more likely to be admitted or to have repeat ED visits, and they experience higher rates of adverse health outcomes after discharge. The risk factors commonly associated with the negative outcomes are age, functional impairment, recent hospitalization or ED use, living alone, and lack of social support. Comprehensive geriatric screening and coordinated discharge planning initiatives designed to improve clinical outcomes in older emergency patients have provided inconclusive results. CONCLUSION: Older ED patients have distinct patterns of service use and care needs. The current disease-oriented and episodic models of emergency care do not adequately respond to the complex care needs of frail older patients. More research is needed to determine the effectiveness of screening and intervention strategies targeting at-risk older ED patients. PMID- 11867976 TI - The prevalence and documentation of impaired mental status in elderly emergency department patients. AB - STUDY OBJECTIVE: We sought to determine the prevalence of mental status impairment in elderly emergency department patients and to assess documentation of and referrals by emergency physicians for mental status impairment after discharge from the ED. METHODS: We performed a prospective, observational study of a convenience sample of 297 patients 70 years or older presenting to an urban teaching hospital ED over a 12-month period. Patients were screened with the Orientation-Memory-Concentration examination for cognitive impairment and the Confusion Assessment Method for delirium. Documentation, dispositions, and referrals were abstracted from chart review. RESULTS: Two hundred ninety-seven of the 337 eligible patients were enrolled. Seventy-eight of the 297 (26%; 95% confidence interval [CI] 21% to 31%) patients had mental status impairment; 30 (10%; 95% CI 7% to 14%) had delirium; 48 (16%; 95% CI 12% to 20%) had cognitive impairment without delirium; 17 (6%; 95% CI 3% to 9%) screened positive on both examinations. Only 22 (28%; 95% CI 19% to 40%) of the 78 patients had any documentation of mental status impairment by the emergency physician. Specific mention of delirium, cognitive impairment, or an acceptable synonym was noted in 13 (17%; 95% CI 9% to 27%). Of 34 (44%; 95% CI 32% to 55%) patients with mental status impairment discharged home, only 6 (18%; 95% CI 7% to 35%) had plans documented by the emergency physician to address impairment. Eleven (37%; 95% CI 20% to 56%) of the 30 patients with delirium were discharged home. Sixteen (70%; 95% CI 47% to 87%) of the 23 patients with cognitive impairment who were discharged home had no prior history of dementia; these patients were less likely to have specialized assistance with care (13%; 95% CI 4% to 27%) than those with known dementia (58%; 95% CI 28% to 85%). CONCLUSION: Impaired mental status is common among older ED patients. Lack of documentation, admission, or referral by emergency physicians suggests a lack of recognition of this important problem. PMID- 11867977 TI - Serum cleaved tau protein levels and clinical outcome in adult patients with closed head injury. AB - STUDY OBJECTIVE: Intracranial injuries (ICI) are associated with high mortality and morbidity. Unfortunately, tools for diagnosis and risk stratification of ICIs are limited in the emergency department setting. We determine the relationship between the presence or absence of a detectable cleaved serum tau protein (tau(c)), ICI, and outcome at hospital discharge in adults with closed head injuries (CHI). METHODS: This was a prospective pilot study of adult patients with CHI presenting to the ED of an urban Level I trauma center. Patients presenting within 10 hours of a CHI who underwent a head computed tomographic (CT) scan were eligible. A blood sample was collected at presentation and the tau(c) level was measured. Initial Glasgow Coma Scale scores and demographic information were recorded. A chart review was performed to determine outcome and final readings on the initial head CT scan. RESULTS: Patients were dichotomized to 1 of 2 groups, those with an ICI (n=17) and those with an isolated skull fracture or no CT abnormality (NICI) (n=11). The 2 groups were similar in regard to demographic composition, mechanism of injury, and coexisting injuries. A tau(c) level of more than 0 was associated with an increased chance of an ICI on the initial head CT scan (odds ratio 11.25; 95% confidence interval [CI] 1.17 to 108.4) and a greater chance of poor outcome, defined as death while in hospital or transfer to a nursing home at hospital discharge (odds ratio 8.17; 95% CI 1.42 to 47). CONCLUSION: A tau(c) level of more than 0 is associated with a greater chance of intracranial injury and poor outcome in patients with CHI. PMID- 11867978 TI - Adverse drug events in emergency department patients. AB - STUDY OBJECTIVE: Adverse drug events (ADEs) have been studied in hospitalized patients. Less is known about this common type of injury in emergency department patients. This study seeks to measure the risks, incidence, severity, and costs of ADEs in an ED population. METHODS: ED charts of visits to a university affiliated tertiary-care ED occurring between March 1 and May 31, 1997, were retrospectively reviewed. The main outcome measures were ADE incidence, severity, and total cost. Visits identified by investigators as containing a suspected ADE were further assessed by using the Naranjo Adverse Drug Reaction (ADR) probability scale. Events judged as probable ADEs (Naranjo ADR probability scale score of >4) were compared with ED control visits best matched by age for disposition, survival, severity, payer, sex, race, age, number of drugs, and total cost. RESULTS: Of 13,602 visits, 13,004 records were available. Three hundred twenty-one had suspected and 217 had probable ADEs (1.7% of evaluable encounters); these were compared with visits by 217 age-matched control patients. Insulin and warfarin were the most commonly responsible drugs. Patients with ADEs were older (mean age 45.1 versus 36.8 years; mean difference 8.3; 95% confidence interval [CI] 3.7 to 12.9), were more often women (odds ratio [OR] 1.48; 95% CI 1.01 to 2.16), took more drugs (mean number of drugs 4.1 versus 1.9; mean difference 2.2; 95% CI 1.7 to 2.8), and were hospitalized more frequently (OR 2.29; 95% CI 1.33 to 3.94) than control patients. CONCLUSION: ADEs encompassed an important segment of ED encounters and annual health care costs. ED screening may provide useful information about the epidemiology of outpatient ADEs. PMID- 11867979 TI - Pediatric orogastric and nasogastric tubes: a new formula evaluated. AB - STUDY OBJECTIVE: We sought to compare the traditional method of determining depth of gastric tube insertion, by measuring from the external landmarks of the nose or mouth, to the earlobe, to the xiphoid process (NEX method), with a graph for determining depth of gastric tube insertion that is based on patient height (graphic method). METHODS: A prospective, randomized, double-blinded study comparing NEX and graphic methods for gastric tube depth of insertion was undertaken. This study included a convenience sample of pediatric emergency department patients in need of gastric intubation. Patients were block randomized, and their gastric tubes were placed to the depth derived from the particular method employed. Alternate depth of insertion was measured on all patients. Abdominal radiographs were used to determine the distance that the end of the tube was from the center of the stomach. RESULTS: Forty-four patients each were in the NEX and graphic groups. The mean distance from the center of the stomach was -1.12 cm (SD 1.36) for the graphic group, compared with 1.31 cm (SD 3.39) for the NEX method. The difference between the 2 methods was 2.43 cm (95% confidence interval [CI] 1.33 to 3.54). Using absolute values, the mean distance from the center of the stomach was 1.26 cm (SD 1.23) for the graphic group compared with 2.60 cm (SD 2.51) for the NEX method. Using these values, the difference between the groups is 1.34 cm (95% CI 0.50 to 2.18). CONCLUSION: When compared with the NEX method, the graphic method demonstrates a significant ability to more consistently and accurately determine the depth of pediatric gastric tube insertion. PMID- 11867980 TI - The role of activated charcoal and gastric emptying in gastrointestinal decontamination: a state-of-the-art review. AB - Gastrointestinal decontamination has been practiced for hundreds of years; however, only in the past few years have data emerged that demonstrate a clinical benefit in some patients. Because most potentially toxic ingestions involve agents that are not toxic in the quantity consumed, the exact circumstances in which decontamination is beneficial and which methods are most beneficial in those circumstances remain important topics of research. Maximum benefit from decontamination is expected in patients who present soon after the ingestion. Unfortunately, many overdose patients present at least 2 hours after taking a medication, when most of the toxin has been absorbed or has moved well into the intestine, beyond the expected reach of gastrointestinal decontamination. Decontamination probably does not contribute to the outcome of many such patients, especially those without symptoms. However, if absorption has been delayed or gastrointestinal motility has been slowed, activated charcoal may reduce the final amount absorbed. The use of activated charcoal in these cases may be beneficial and is associated with few complications. Therefore, administration of activated charcoal is recommended as soon as possible after emergency department presentation, unless the agent and quantity are known to be nontoxic, the agent is known not to adsorb to activated charcoal, or the delay has been so long that absorption is probably complete. The use of gastric emptying in addition to activated charcoal has generated intense debate. Several large comparative studies have failed to demonstrate a benefit of gastric emptying before activated charcoal. Because complications of such 2-step decontamination include a higher rate of intubation, aspiration, and ICU admission, gastric emptying in addition to activated charcoal cannot be considered the routine approach to patients. However, there are several infrequent circumstances in which the data are inadequate to accurately assess the potential benefit of gastric emptying in addition to activated charcoal: symptomatic patients presenting in the first hour after ingestion, symptomatic patients who have ingested agents that slow gastrointestinal motility, patients taking sustained release medications, and those taking massive or life threatening amounts of medication. These circumstances represent only a small subset of ingestions. In the absence of convincing data about benefit or lack of benefit of gastric emptying for these patients, individual physicians must act on a personal valuation: Is it better to use a treatment that might have some benefit but definitely has some risk or not to use a treatment that has any risk unless there is proven benefit? PMID- 11867981 TI - Coping with medical mistakes and errors in judgment. AB - Attention has recently been focused on medical errors as a cause of morbidity and mortality in clinical practice. Although much has been written regarding the cognitive aspects of decision making and the importance of systems management as an approach to medical error reduction, little consideration has been given to the emotional impact of errors on the practitioner. Evidence exists that errors are common in clinical practice and that physicians often deal with them in dysfunctional ways. However, there is no general acknowledgment within the profession of the inevitability of medical errors or of the need for practitioners to be trained in their management. This article focuses on the affective aspects of physician errors and presents a strategy for coping with them. PMID- 11867982 TI - Evidence-based emergency medicine. Improving the dissemination of systematic reviews in emergency medicine. PMID- 11867983 TI - Evidence-based emergency medicine. Use of thrombolytic therapy in patients with acute ischemic stroke. PMID- 11867984 TI - Evidence-based emergency medicine. Angioplasty versus intravenous thrombolysis for acute myocardial infarction. PMID- 11867985 TI - Evidence-based emergency medicine. How to find evidence when you need it, part 1: databases, search programs, and strategies. PMID- 11867986 TI - Thallium and arsenic poisoning in a small midwestern town. AB - Thallium and arsenic have been used as a means of criminal poisoning. Although both manifest characteristically with peripheral neuropathies, thallium is associated with alopecia and arsenic with gastrointestinal symptoms. We describe the symptoms, physical findings, diagnostic test results, and outcomes in a group of men poisoned with thallium and arsenic. Seven patients had evidence of elevated thallium levels, and 2 patients had elevated arsenic and thallium levels. The most commonly reported symptoms included myalgias, arthralgias, paresthesias, and dysesthesias. Five patients developed alopecia. All patients with symptoms and peripheral neuropathies had characteristic blackening of their hair roots. Initially treated with dimercaptosuccinic acid, patients were switched to multiple-dose activated charcoal after testing revealed thallium poisoning. By 6 months, all patients' symptoms and peripheral neuropathies improved, but 5 patients had ongoing psychiatric problems. Thallium remains a means of criminal poisoning and should be considered in any patient with a rapidly progressing peripheral neuropathy with or without alopecia. PMID- 11867988 TI - Residents as teachers in the emergency department. PMID- 11867987 TI - Mercuric oxide poisoning treated with whole-bowel irrigation and chelation therapy. AB - Most reported cases of inorganic mercury poisoning are from mercuric chloride. We report a case of mercuric oxide (HgO) powder ingestion. A 31-year-old man presented to an emergency department after ingestion of approximately 40 g of HgO. Soon after ingestion, he developed nausea, vomiting, and abdominal cramping. Abdominal radiograph revealed densely radiopaque material in the stomach. Gastrointestinal decontamination was accomplished with activated charcoal and whole-bowel irrigation with polyethylene glycol solution (Golytely) for 24 hours until repeat abdominal radiographs no longer demonstrated the substance in the gastrointestinal tract. He was also chelated with British anti-Lewisite for 5 days, followed by succimer for 10 days. He had markedly elevated urine and blood mercury levels after ingestion, but except for a mildly depressed serum bicarbonate (19 mEq/L), his chemistry results remained normal including blood urea nitrogen and creatinine. He had an uncomplicated hospital course and remained asymptomatic at 6 months postingestion. Despite elevated urine and blood mercury levels after ingestion of HgO, our patient did not develop the end-organ toxicity typical of inorganic mercury poisoning. PMID- 11867989 TI - The management of occupational exposures to blood and body fluids: revised guidelines and new methods of implementation. PMID- 11867990 TI - Abridged version of the updated US Public Health Service guidelines for the management of occupational exposures to hepatitis B virus, hepatitis C virus, and human immunodeficiency virus and recommendations for postexposure prophylaxis. PMID- 11867991 TI - Brain cramp: the emergency physician's worst nightmare. PMID- 11867992 TI - Panic. PMID- 11867993 TI - Headaches from practice guidelines. PMID- 11867994 TI - Missed delirium in older emergency department patients: a quality-of-care problem. PMID- 11867995 TI - Rule out TBI? Serum markers for traumatic brain injury. PMID- 11867996 TI - Dealing with failure: the aftermath of errors and adverse events. PMID- 11867997 TI - Preventing complications of adenosine administration. PMID- 11867998 TI - Two cases of rattlesnake envenomation with delayed coagulopathy. PMID- 11867999 TI - Use of computed tomographic scans for patients with minor head injury. PMID- 11868000 TI - Shift work and physician performance. PMID- 11868001 TI - Procedural success and complications of large-scale screening colonoscopy. AB - BACKGROUND: Indirect evidence and modeling analyses suggest that colonoscopy may be the most cost-effective way to screen the average-risk population for colorectal neoplasia. However, the success and safety of primary colonoscopic screening has not been prospectively evaluated in a multicenter trial. METHODS: Asymptomatic subjects age 50 to 75 years who had not undergone examination of the colon within 10 years were recruited from the general medicine clinics of 13 Department of Veterans Affairs Medical Centers. Eligible patients underwent colonoscopy by study coinvestigators, at which time all polyps were measured, photographed, and removed. Patients were contacted at 24 hours and 1 week to track procedure-related complications. RESULTS: Primary screening colonoscopy was performed in a cohort of 3196 asymptomatic subjects. A "good" preparation was reported in 81% of patients, and colonoscopy to the cecum was successful in 97.2% of cases. Mean insertion time to the cecum and total procedure times were 10.5 (8.7) and 30.6 (19.1) minutes, respectively. No preprocedural patient characteristics were identified that were predictive of an incomplete procedure. At least one polyp was resected in 1672 patients. There was no perforation and no death attributed to colonoscopy. Major morbidity considered to be definitely related to colonoscopy occurred in 9 of 3196 procedures (0.3%): lower GI bleeding requiring intervention (6), myocardial infarction and/or cerebrovascular accident (2), and thrombophlebitis (1). In subjects undergoing only diagnostic procedures, the major complication rate was 0.1%. CONCLUSIONS: Screening colonoscopy can be performed in multiple centers with a high degree of success and safety in large numbers of asymptomatic, average-risk men. PMID- 11868002 TI - Flexible sigmoidoscopy: the patients' perception. AB - BACKGROUND: Anxiety, discomfort, and high levels of concern can affect patient attitude toward endoscopic procedures as well as compliance and adherence to current recommendations for the examination. This study evaluated how patients perceive flexible sigmoidoscopy. METHODS: A prospective study was conducted at two sites of 764 patients presenting for GI endoscopy (flexible sigmoidoscopy 175, colonoscopy 384, EGD 205). Before these procedures, patients rated their anticipated (preprocedure) difficulty and degree of concern for each of 30 specific concerns. After the procedure, the patients rated their actual (postprocedure) difficulty. Patients' levels of concerns and difficulty were compared before and after the procedure and among the 3 procedure types. RESULTS: Before the procedure, patients who were to undergo flexible sigmoidoscopy had fewer concerns and lower scores for the severity of the concerns than did patients having colonoscopy or EGD. After the procedure, patients who had flexible sigmoidoscopy rated it as more difficult than patients who had colonoscopy or EGD. Patients who had colonoscopy and EGD graded their actual difficulty as less than their anticipated difficulty. However, patients who had flexible sigmoidoscopy rated the actual difficulty the same as the anticipated difficulty. CONCLUSIONS: Although patients have fewer concerns regarding flexible sigmoidoscopy than for the other endoscopic procedures, after the procedure they rate sigmoidoscopy as more difficult compared with postprocedure ratings by patients who underwent colonoscopy or EGD. This suggests that it may be necessary to change the perception of flexible sigmoidoscopy as being the best tolerated of the endoscopic procedures. PMID- 11868003 TI - Utilization of health care resources for low-risk patients with acute, nonvariceal upper GI hemorrhage: an historical cohort study. AB - BACKGROUND: Adults hospitalized with acute, nonvariceal upper GI hemorrhage can be accurately stratified according to their risk of subsequent adverse outcomes by using the Rockall score. Low-risk patients (Rockall score less-than-or-equal 2) may be candidates for early discharge. METHODS: Cases were identified with ICD 9-CM codes for calendar years 1997 and 1998. Medical record data to determine patient Rockall risk score, health care resource utilization, and adverse outcomes were abstracted with standardized forms. RESULTS: Fifty-three of 175 (30%) cases had Rockall scores < or =2. As predicted, those patients with Rockall scores < or =2 had a low risk of adverse outcomes with only 2 of 53 (4%) meeting criteria for recurrent bleeding as defined by the "Rebleed" variable, and no mortality. These low-risk patients had a mean hospital stay of 2.6 plus minus 2.1 days; 49% were admitted to an intermediate or intensive care unit bed and 57% were given H2 receptor antagonists intravenously. CONCLUSIONS: The proportion of patients admitted with acute, nonvariceal, upper GI hemorrhage with Rockall Scores < or =2 was substantial. Adverse outcomes were rare. In contrast, the level of health care resource utilization appeared high. The Rockall score has potential as a clinically based concurrent decision rule to improve the quality of care by finding those patients less likely to require intensive health care services. PMID- 11868004 TI - A resource utilization projection study of EUS. AB - BACKGROUND: EUS has emerged as standard practice with respect to the diagnosis and staging of GI malignancies. Whether currently available resources are sufficient to meet the potential need for EUS is uncertain. This study examines the hypothetical demand for EUS in the United States. METHODS: EUS cases performed at 3 centers with well-established expertise in EUS in 1997 were retrospectively reviewed and trends were extrapolated to national cancer statistics. Indications for EUS fell into 3 categories: (1) diagnosis/staging of esophageal, gastric, pancreatic, or rectal cancers (established indications); (2) suspected GI malignancy (obligate "rule out"); and (3) "other" (emerging indications). Hypothetical total numbers of cases in which EUS could be performed in the United States were calculated taking into consideration the expected number of GI malignancies for which EUS would be appropriate (based on cancer statistics for 2000), the fraction unresectable by CT, the fraction of elderly nonsurgical candidates, proportionate "rule out" cases, as well as "other" emerging indications. RESULTS: The calculated hypothetical number of cases (United States) in which EUS would be indicated is 79,568 per year (10,287 esophagus, 10,666 stomach, 23,069 pancreas, and 35,546 rectal). If "other" indications remained constant at 12%, there would potentially be 89,116 EUS procedures performed per year, with a conservative estimate of 79,572 per year. CONCLUSIONS: This model suggests that currently available EUS resources are not sufficient to meet hypothetical demand. Future considerations include the number of endoscopy units in which EUS is performed, the capacity of individual units, and the implications for training programs in the United States. PMID- 11868005 TI - Endoscopic implantation of a biopolymer in the lower esophageal sphincter for gastroesophageal reflux: a pilot study. AB - BACKGROUND: GERD is the most frequent disorder of the esophagus. Endoscopic minimally invasive treatment is desirable. However, the results of injection techniques have been disappointing. METHODS: A pilot study was conducted in patients with GERD, who required continuous therapy with a proton pump inhibitor, in which ethylene-vinyl-alcohol was injected into the muscle of the gastric cardia. Primary endpoints were the safety of the procedure, the effect on lower esophageal sphincter pressure and the stability of the injected material. A secondary endpoint was the effect on heartburn score after discontinuation of treatment with a proton pump inhibitor. RESULTS: Ethylene-vinyl-alcohol injection into the cardia resulted in circular diffusion of the product in 10 of 15 cases, suggesting that implantation into the muscle is feasible. Lower esophageal sphincter pressure was increased in 13 of 15 cases at 1 month and was sustained at a median follow-up of 6 months (range 4-12 months). Mean plus minus SEM of lower esophageal sphincter pressures (15 patients) were 12.2 plus minus 0.9, 18.7 plus minus 1.5 (p = 0.001 at baseline), and 16.7 plus minus 1.3 mm Hg (p = 0.038 from baseline) at, respectively, baseline, 1 month follow-up, and final follow up. There was also a sustained reduction in heartburn score (off proton pump inhibitor) (3.40 plus minus 0.13 vs. 1.53 plus minus 0.24 and 1.87 plus minus 0.26 at baseline vs. 1 month and final follow-up, respectively; p < 0.01). Nine of the 15 patients had more than 50% of the injected material in place at second follow-up (at 6 months for 8 patients; at 12 months for 1 patient). In only 2 patients was there loss of more than 75% of injected ethylene-vinyl-alcohol. Persistence of greater than 50% of the material was associated with achievement of a circular injection. Only 4 patients had to resume therapy with a proton pump inhibitor. Mild retrosternal discomfort was observed in 8 patients; this disappeared in all cases after a maximum of 3 days. CONCLUSIONS: Ethylene-vinyl alcohol implantation in the muscle of the cardia is feasible and safe. It leads to a sustained increase in resting lower esophageal sphincter pressure. This is associated with a sustained improvement in heartburn score for patients who previously required continuous therapy with a proton pump inhibitor. PMID- 11868006 TI - The influence of mutation site and age on the severity of duodenal polyposis in patients with familial adenomatous polyposis. AB - BACKGROUND: The present study was undertaken to identify factors affecting the severity of the duodenojejunal polyposis in patients with familial adenomatous polyposis. METHODS: Duodenojejunal polyposis was evaluated in 41 consecutive patients with familial adenomatous polyposis (mean age 41 years, range 21-63 years), 33 (80%) with known APC mutation, by using a standardized endoscopic protocol. The severity of the polyposis was graded with the Spigelman scoring system (0-12 points), the Spigelman score/age ratio, and the presence or absence of advanced adenomas (>1 cm in diameter and/or high-grade dysplasia). RESULTS: The Spigelman score (median 8, range 3-12) was higher in patients older than 50 years (median 10, range 3-12) as compared with younger patients (median 7.5, range 3-11; p = 0.043). A significant association between age and the presence of advanced adenomas was also observed. Patients with a mutation in the central part of the APC gene (codons 279-1309) had a higher Spigelman score and Spigelman score/age ratio as compared with patients with other mutations: median Spigelman score/age ratio 0.21 (range 0.14-0.40) versus 0.10 (range 0.06-0.20) (p < 0.001). CONCLUSIONS: Older age and APC mutation in the central part of the gene are risk factors for the development of severe duodenojejunal polyposis. PMID- 11868007 TI - Epinephrine versus epinephrine plus fibrin glue injection in peptic ulcer bleeding: a prospective randomized trial. AB - BACKGROUND: Peptic ulcer bleeding remains a disease with considerable morbidity and mortality. Epinephrine is the most widely used endoscopic injection agent, but bleeding recurs in 20% of high-risk cases. Fibrin glue might be an ideal injection agent, based on its physiologic properties, despite its demanding injection technique and high cost. The aim of this study was to determine whether the injection of fibrin glue in combination with epinephrine improves outcome for patients at high risk of recurrent bleeding. METHODS: Patients were prospectively randomized to injection of epinephrine alone (n = 70) or epinephrine plus fibrin glue (n = 65). Endoscopy was repeated daily until the ulcer base was clean. All patients were treated with high-dose omeprazole. RESULTS: Initial hemostasis was 100% in both groups. There was no significant overall difference in rates of recurrent bleeding (24.3% and 21.5%, respectively, for epinephrine and epinephrine plus fibrin). When patients were stratified according to Forrest criteria, no significant difference could be found, although there was a trend toward less recurrent bleeding after fibrin injection of actively bleeding ulcers. There was no significant difference in the proportions of patients who required surgery (10% and 6%, respectively, for epinephrine and epinephrine plus fibrin). Mortality was the same (3%) in each group. CONCLUSIONS: Adding fibrin glue to epinephrine for injection treatment of bleeding peptic ulcers does not improve outcome. Fibrin glue might be of some value in selected cases. PMID- 11868008 TI - "Tannenbaum" Teflon stents versus traditional polyethylene stents for treatment of malignant biliary stricture. AB - BACKGROUND: Premature stent clogging is the major limitation with plastic stents used in the treatment of malignant biliary structures. A pilot study suggested improved duration of patency of the Tannenbaum stent compared with polyethylene stents. The aim of this prospective, multicenter randomized trial was to compare the Tannenbaum Teflon stent with a conventional polyethylene endoprosthesis (Cotton-Leung biliary stent set) for the treatment of malignant biliary strictures. METHODS: Patients over age 18 years with symptoms caused by nonhilar malignant biliary strictures were enrolled. Patients were randomized to receive a 10F Tannenbaum or polyethylene stent after a guidewire was passed beyond the stricture. One hundred six patients (mean age 72 years and 71 years, respectively) were enrolled (54 Tannenbaum, mean age 72 years; 52 polyethylene, mean age 71 years). RESULTS: Tannenbaum and polyethylene stent placement was successful in, respectively, 100% and 96% of procedures without complications. The mean (SD) 90-day stent patency of the Tannenbaum stent was 67% (7%) compared with 73% (7%) for the polyethylene stents. CONCLUSIONS: The present study demonstrated no difference in ease of implantation or stent patency between Tannenbaum and polyethylene stents. PMID- 11868010 TI - Polyurethane-covered metal stent for management of distal malignant biliary obstruction. AB - BACKGROUND: The efficacy of polyurethane-covered metal expandable stents for treatment of neoplastic distal biliary obstruction was analyzed. METHODS: Twenty one patients with unresectable malignant tumors involving the mid to distal bile duct who were seen with obstructive jaundice were consecutively enrolled. Eighteen patients underwent endoscopic implantation of a polyurethane-covered metal stent and 3 patients underwent transhepatic insertion because of duodenal obstruction. The stent patency, complications, and patient survival were analyzed. RESULTS: Effective biliary decompression was achieved in all patients. Adverse events were minor: mild pancreatitis (1) and cholecystitis (2) occurred within 7 days of stent insertion. Mean survival and stent patency were 233 days and 206 days, respectively. Stent occlusion occurred in 3 patients (14%) after a mean of 188 days. Tumor ingrowth through the stent mesh was not observed. However, 2 stents were occluded by tumor overgrowth and 1 by compacted alimentary debris. CONCLUSIONS: A polyurethane membrane may prevent tumor ingrowth and reduce the occlusion rate for expandable metal stents implanted in patients with malignant obstruction of the distal bile duct. PMID- 11868009 TI - Features of malignant biliary obstruction affecting the patency of metallic stents: a multicenter study. AB - BACKGROUND: Although placement of a metallic stent is an established therapeutic option for the palliation of patients with malignant biliary obstruction, it remains unclear which stricture-related or stent-related factors influence stent patency. METHODS: Metallic stents were inserted endoscopically in 68 patients (38 men, 30 women; mean age 70.2 plus minus 8.5 years) with malignant biliary obstruction. Patency rates were prospectively analyzed according to malignant stricture characteristics, including length, morphologic type, and degree of narrowing. Furthermore, patient age, initial serum bilirubin level, length of stent, time to adequate expansion, and the location of the distal end of the stent were evaluated as possible factors affecting the stent patency. Stent patency was assessed by using the survival analysis of the Kaplan-Meier estimation and Cox regression analysis. RESULTS: Median overall stent patency was 231 days (range 27-379 days) and the overall rate of stent occlusion 41.2% (28/68). The causes of occlusion were tumor ingrowth in 23 patients (33.8%), distal overgrowth in 3 (4.4%), proximal overgrowth in 1 (1.5%), and encrustation with sludge in 1 patient (1.5%). No significant differences in patency rates according to patient age, initial serum bilirubin level, primary tumor type, length and morphologic type of stricture, and length and location of the distal end of the stent were found. Cox regression analysis demonstrated that the degree of narrowing assessed by cannula or guidewire passage and the time to adequate expansion of the stent were independent factors associated with stent patency. CONCLUSION: Early expansion of the stent and easy passage of larger-caliber instruments through the stricture were favorable factors for long-term patency of the Wallstent. PMID- 11868011 TI - Treatment with argon plasma coagulation reduces recurrence after piecemeal resection of large sessile colonic polyps: a randomized trial and recommendations. AB - BACKGROUND: Recurrence is frequent after piecemeal snare resection of large sessile colorectal polyps. The aim of this study was to evaluate the safety and efficacy of argon plasma coagulation (APC) in preventing recurrence when applied to the edge and base of the polypectomy site after apparently complete piecemeal resection. METHODS: Patients with large (>1.5 cm) sessile polyps removed by piecemeal snare cautery were placed into 2 groups. The first consisted of patients with polyps believed by the endoscopist to be completely excised. These patients were randomized to either no further therapy (control) or to APC of the rim and any residual mucosal or submucosal tissue in the base of the polypectomy site. The second group comprised patients in whom polyps, as judged by the endoscopist, were incompletely excised by snare polypectomy; APC was routinely applied without randomization to all visible remaining adenomatous tissue. Follow up colonoscopy was performed within 3 months and 1 year; biopsy specimens were taken routinely from the resection site and further polypectomy was performed as indicated. RESULTS: There were fewer recurrences after APC in the randomized group (1/10 APC, 7/11 no APC; p = 0.02). In the group with initial incomplete snare polypectomy, recurrence was detected at 3 months in 6 of 13 despite APC. One patient was hospitalized with abdominal pain and minor rectal bleeding but required no intervention. There were no other episodes of significant late bleeding caused by piecemeal polypectomy. One patient was referred for surgery after unsuccessful endoscopic management. CONCLUSIONS: In patients with apparent complete endoscopic snare resection of large adenomas, postpolypectomy application of APC reduces adenomatous recurrence. PMID- 11868012 TI - Long-term results of endoscopic management of pancreas divisum with recurrent acute pancreatitis. AB - BACKGROUND: The long-term efficacy of endoscopic treatment in pancreas divisum is controversial. This study evaluated the long-term results of dorsal duct stent insertion and endoscopic sphincterotomy of the minor papilla in patients presenting with recurrent acute pancreatitis or chronic pain. METHODS: Pancreas divisum was diagnosed in 175 patients between 1980 and 1998. Twenty-four patients seen with recurrent acute pancreatitis without underlying chronic calcifying pancreatitis or significant alcohol consumption were included in this study with a follow-up of at least 24 months. Eight were treated by sphincterotomy of the minor papilla alone, and 16 underwent dorsal duct stent insertion for a median duration of 8 months. RESULTS: The median duration of follow-up after endoscopic management was 39 months (range 24-105; interquartile range 40.5). All patients had recurrent acute pancreatitis before endoscopic treatment during a median period of 5 years. At the end of the follow-up there were only 2 recurrences of acute pancreatitis (p < 0.01). The number of patients with chronic pain before endoscopic treatment and at the end of follow-up decreased from 20 of 24 (83%) to 7 of 24 (29%) without reaching statistical significance. The 25% recurrence rate was estimated at 50 months by Kaplan-Meier analysis. Nine patients presented with a dilated dorsal duct before endoscopic treatment. After stent insertion, dorsal duct dilatation appeared in all 16 patients who underwent stent placement, and pancreatic duct stenosis developed in 3. Four patients (19%) required further treatment for pain recurrence or acute pancreatitis, with surgical procedures being performed in 2 cases. Complications occurred in 9 of 24 patients (38%), mainly acute pancreatitis or stenosis of the minor papilla. All complications except one were managed conservatively. Complications seemed to be less frequent after minor papilla sphincterotomy than after pancreatic stent insertion (25% vs. 44%). CONCLUSION: In patients with pancreas divisum, both dorsal duct stent insertion and minor papilla sphincterotomy decrease the rate of recurrent acute pancreatitis, whereas relief of chronic pain was less obvious. PMID- 11868013 TI - Comparative cytotoxicity of low-osmolar nonionic and high-osmolar ionic contrast media to dog gallbladder epithelial cells. AB - BACKGROUND: Most studies of the adverse effects of x-ray contrast media used in ERCP have focused on post-ERCP pancreatitis. However, the biliary epithelial cells are also exposed to contrast media during ERCP and acute cholangitis is also a serious complication of ERCP. The present study compared the cytotoxicity with gallbladder epithelial cells of ionic and nonionic contrast agents. METHODS: A high-osmolar ionic contrast agent (meglumine ioxithalamate) and a low-osmolar nonionic contrast agent (iopromide) were tested. Monolayer cell cultures of dog gallbladder epithelial cells were used. The cells were exposed to the 2 contrast agents with increasing iodine concentration and osmolality for 2 days. Cell number, S-phase fraction, aneuploidy, and supernatant LDH activities were measured each day. RESULTS: Cell growth was more severely inhibited by ioxithalamate than iopromide (p < 0.05) and strongly dependent on the osmolality of contrast agent. The cytostatic effect estimated by S-phase fraction was more pronounced for ioxithalamate. Chromosomal damage determined by aneuploidy was more frequently detected with ioxithalamate. CONCLUSIONS: High-osmolar ionic contrast media are more cytotoxic than low-osmolar nonionic contrast media to gallbladder epithelial cells. Animal and clinical studies are needed to estimate the clinical implications of these findings. PMID- 11868014 TI - Is endoscopic follow-up needed in pediatric patients who undergo surgery for GERD? AB - BACKGROUND: This study evaluated the role of endoscopy in the postoperative management of pediatric patients who undergo fundoplication for GERD. METHODS: Medical records of 109 otherwise healthy children who underwent operation for GERD from 1979 to 1996 were reviewed. Patients with respiratory symptoms or esophageal stenosis were excluded. All patients underwent endoscopic surveillance with endoscopy being performed in the early (within 1 year) and late (between 1 and 2 years) postoperative periods. Specifically evaluated were the appearance of the wrap and evidence of esophagitis. The risk of a recurrence of esophagitis based on wrap appearance and the presence of clinical symptoms in patients with endoscopic evidence of esophagitis were also evaluated. RESULTS: At early endoscopy 3 patients with an intact wrap and 8 with a defective wrap had esophagitis (not significant). At late endoscopy, 5 patients with an intact wrap and 17 with a defective wrap had esophagitis (p < 0.05). CONCLUSIONS: An intact wrap does not prevent recurrence of GERD. Such an occurrence is even more likely when endoscopy demonstrates a defective wrap. For all patients who have undergone fundoplication, endoscopic evaluation at 1 to 2 years is recommended to detect esophagitis in the absence of symptoms so treatment can be initiated before symptoms occur. PMID- 11868015 TI - Efficacy, safety, and clinical outcomes of endoscopic mucosal resection: a study of 101 cases. AB - BACKGROUND: Endoscopic mucosal resection (EMR) is an alternative to surgery for removal of superficial neoplastic lesions of the GI tract. The aim of this study was to assess the safety, efficacy, and clinical outcomes of EMR. METHODS: Data from consecutive EMR procedures performed by using suction cap-assisted and/or saline solution-assisted snare resection techniques over a 45-month period were reviewed retrospectively. EUS was performed before EMR in the majority of cases. Immediate and delayed complications were recorded. Survival was assessed in patients with carcinoma or high-grade dysplasia on final histopathology in whom EMR achieved complete resection. RESULTS: One hundred one lesions were removed by EMR in 92 patients. Indications were adenoma (67%), high-grade dysplasia (13%), intramucosal carcinoma (11%), and lesions of uncertain histopathology (10%). Locations were esophagus 19%, stomach 14%, duodenum 27%, rectum 12%, and colon 29%. Suction cap-assisted technique was used in 26% and saline solution-assisted polypectomy in 74% of cases. Complete resection was achieved in 89%. For complete resection, 17% required more than 1 session. Post-EMR histopathology was adenoma 47%, high-grade dysplasia 13%, carcinoma 16%, carcinoid 3%, benign 19%, and low grade dysplasia 3%. EMR resulted in upgrading of histopathologic staging to carcinoma or high-grade dysplasia in 44%. Bleeding was the only complication (early 16, delayed 6). The median cancer-free survival in patients with adenocarcinoma who underwent complete resection by EMR was 27 months (interquartile range: 17-28 months). CONCLUSION: EMR achieves complete resection in a majority of patients but is associated with a higher risk of bleeding compared with standard polypectomy. EMR changes pathologic stage in a significant number of patients. Survival data are encouraging, but long-term follow-up studies are needed. PMID- 11868016 TI - Evaluation of the pancreaticobiliary ductal systems by intraductal US. PMID- 11868017 TI - Rectal varices caused by thrombosis of intra-abdominal vessels. PMID- 11868018 TI - Xanthoma of the colon. PMID- 11868019 TI - Splenic, portal, and superior mesenteric vein confluens at EUS. PMID- 11868020 TI - Ileocecal ulcer caused by chemotherapy. PMID- 11868021 TI - A new stereoscopic endoscopy system: accurate 3-dimensional measurement in vitro and in vivo with distortion-correction function. AB - BACKGROUND: The purpose of this study was to evaluate the accuracy and reproducibility of measurements obtained with a new 3-dimensional stereoscopic endoscopy system in vitro and in vivo. METHODS: The system includes an electronic stereoscopic endoscope and a personal computer. After correction of barrel distortion by an image-processing technique, 3-dimensional data are computed with triangulation. Initially, graph paper and models of gastric lesions, both elevated and depressed, were used as objects. For evaluating accuracy and reproducibility, the absolute error (/E/) and the coefficient of variation (CV) were calculated: /E/ = average value of absolute value (measured value minus sign true value)/true value. Then, the sizes of 50 gastric lesions were measured by 2 endoscopists, one experienced and the other inexperienced, and the findings were compared with measurements made radiographically. RESULTS: The maximum for /E/ and for the CV for the distance on the graph paper were, respectively, 5.6% and 7.0%. The /E/ and CV for the diameter and for the height of the elevated model were, respectively, 5.9% and 5.7% for the former and, respectively, 16.8% and 16.5% for the latter. The /E/ and CV for the area of the depressed model were, respectively, 9.0% and 1.6%. The correlation coefficients between the sizes of lesions measured by stereoscopic endoscopy and radiographically were 0.91: 99% CI [0.81, 1.00], and 0.98: 99% CI [0.85, 1.00], respectively, for the experienced and inexperienced endoscopists. CONCLUSIONS: The new stereoscopic endoscopy system is useful for measuring gastric lesion size in clinical practice. PMID- 11868022 TI - Photodynamic effects on rabbit auricular veins after photosensitization with porfimer sodium: Implications of the results with respect to the treatment of esophageal varices with photodynamic therapy. AB - BACKGROUND: There are numerous clinical applications for photodynamic therapy in the GI tract. The principal reason for the wide variety of lesions amenable to photodynamic therapy is the ability to treat large areas of mucosa without the need for complete visualization. This report describes observed hemodynamic and histologic changes in rabbit auricles after photodynamic therapy and the feasibility of photodynamic therapy for esophageal varices. METHODS: Porfimer sodium and an argon-dye laser (630 nm, 300 mW/cm(2)) were used. Twenty rabbits were grouped according to porfimer sodium dose: group 1 (2.0 mg/kg, n = 10); group 2 (1.0 mg/kg, n = 6); group 3 (0.2 mg/kg, n = 4). Rabbit auricular veins were classified according to time duration of laser illumination: V(0), no illumination; V(5), 5 minutes; V(10), 10 minutes; V(15), 15 minutes. Hemodynamic changes were observed with a laser Doppler blood flow meter. Histologic changes were evaluated by light microscopy. RESULTS: For groups 1 and 2, there was a significant decrease in blood flow for V(15) after photodynamic therapy, but not in group 3. There was a significant difference in the grade of thrombus between V(5) and V(15) in groups 1 and 2, and between V(10) and V(15) in group 2. There was a significant difference in the grade of venous dilation (congestion) for V(15) between groups 1 and 3 (p < 0.05, Kruskal-Wallis test). CONCLUSIONS: Endoscopic photodynamic therapy could possibly improve the outcome for endoscopic treatment of esophageal varices beyond that achieved by sclerotherapy or band ligation alone. PMID- 11868023 TI - Intramural incision technique. AB - BACKGROUND: Therapeutic ERCP is technically complex. A variety of techniques can be used to achieve biliary access when direct cannulation is difficult. Pre-cut papillotomy can provide immediate access, but this technique is associated with an increased risk of complications. METHODS: An intramural incision technique is described that uses the false tract created with a guidewire to place a papillotome though the intramural portion of the papilla and unroof the biliary orifice. OBSERVATIONS: Biliary access was achieved in 6 consecutive patients in whom access could not be obtained with either a papillotome or guidewire. No major complication occurred. CONCLUSIONS: By virtue of the incision depth, the intramural incision technique offers a safer approach to biliary access than conventional pre-cut techniques. The technique is simple and will be useful when there is aberrant passage of a guidewire. PMID- 11868024 TI - Sphincter of Oddi dysfunction in patients addicted to opium: an unrecognized entity. AB - BACKGROUND: Opiate-induced sphincter of Oddi dysfunction (SOD) as a clinical entity has not been described. METHODS: Eight chronic opium addicts (all men, mean age 61.3 years, mean duration of addiction 24.75 years) presenting with pancreatobiliary pain and a dilated bile duct with or without dilated pancreatic duct on abdominal US were studied. All patients underwent ERCP and biliary sphincterotomy. In addition, pancreatic sphincterotomy was performed in 4 patients with a dilated pancreatic duct. OBSERVATIONS: At ERCP, the bile duct was dilated in 8 and pancreatic duct in 4 patients. There was delayed drainage of contrast (>45 minutes) from the bile duct in all 7 patients studied, whereas delayed drainage from the pancreatic duct (>9 minutes) was incidentally observed in 3 patients. In 6 patients followed after sphincterotomy for at least 2 years, there was marked relief of symptoms. Transabdominal US at 2 years follow-up revealed a normal bile duct in 5 and persistent albeit minimal dilatation in 1 patient. Acute pancreatitis developed in 4 patients after ERCP and sphincterotomy, which was fatal in one. No patient had any abnormality in the gallbladder on initial or follow-up transabdominal US. CONCLUSION: SOD in opium addicts is a distinct clinical entity, mainly seen in men in this population, that is characterized by a long history of opium addiction and the absence of prior cholecystectomy or associated gallstone disease. Most patients are seen with the classic clinical picture of SOD with marked long-term improvement in symptoms after endoscopic sphincterotomy. PMID- 11868025 TI - The difficult polypectomy: description of a new dual-endoscope technique. PMID- 11868026 TI - Diagnostic laparoscopy in radiation-induced liver disease. PMID- 11868027 TI - Preoperative chemoradiation therapy after placement of a self-expanding metal stent in a patient with an obstructing rectal cancer: clinical and pathologic findings. PMID- 11868028 TI - Retroflexion in the duodenum for evaluation of duodenal bulb lesions. PMID- 11868029 TI - Gastric echinostomiasis diagnosed by endoscopy. PMID- 11868030 TI - Ischemic colitis complicating aortic dissection. PMID- 11868031 TI - Submucosal tumor-like elevated lesion after autoamputation of a colonic polyp. PMID- 11868032 TI - EUS: past, present, and the future of endoscopy. PMID- 11868033 TI - Screening for GI cancer and payment mechanisms. PMID- 11868034 TI - The clinical usefulness of secretin-enhanced magnetic resonance pancreatography in patients with pancreas divisum and idiopathic acute pancreatitis. PMID- 11868035 TI - Mirizzi's syndrome after cholecystectomy. PMID- 11868036 TI - Endoscopic removal of gastric ectopic pancreas for histologic diagnosis and treatment. PMID- 11868037 TI - Time-to-event analysis. PMID- 11868038 TI - Should reperfusion strategies in myocardial infarction be modified for the very elderly? PMID- 11868039 TI - To cath or not to cath? That used to be the question. PMID- 11868040 TI - Aspirin: redundant in users of nonaspirin, nonsteroidal antiinflammatory agents? PMID- 11868041 TI - Vascular dysfunction and heart failure: epiphenomenon or etiologic agent? AB - Endothelial function plays a key role in the local regulation of vascular tone. Alterations in endothelial function may result in impaired release of endothelium derived relaxing factors or increased release of endothelium-derived contracting factors. Heart failure may impair endothelial function by means of reduced synthesis and release of nitric oxide (NO) or by increased degradation of NO and increased production of endothelin-1. Endothelial dysfunction may worsen heart function by means of peripheral effects, causing increased afterload and central effects such as myocardial ischemia and inducible nitric oxide synthase (iNOS) induced detrimental effects. Evidence from clinical studies has suggested that there is a correlation between decreased endothelial function and increasing severity of congestive heart failure (CHF). Treatments that improve heart function may also improve endothelial dysfunction. The relationship between endothelial dysfunction and heart failure may be masked by the stage of endothelial dysfunction, the location of vessels being tested, and the state of endothelial-dependent vasodilatation response. PMID- 11868042 TI - The renal dopaminergic system, neurohumoral activation, and sodium handling in heart failure. AB - BACKGROUND: Dopamine of renal origin exerts natriuretic and diuretic actions by activating specific receptors located in the renal proximal tubular epithelial cells. Heart failure (HF) is accompanied by activation of several neurohumoral systems. The interaction of these systems with the renal dopaminergic system and its effect on sodium handling in HF are not clarified. METHODS AND RESULTS: We studied 13 patients with decompensated New York Heart Association class III/IV HF and 17 sex- and age-matched patients with mild to moderate stable class I/II HF. We measured plasma catecholamines, aldosterone, type B natriuretic peptide (BNP), sodium, creatinine (UCr), and 24-hour urinary excretion of sodium, UCr, levo-3,4 dihydroxyphenylalanine (L-DOPA), 3-o -methyldopa, dopamine and its metabolites, 3,4-dihydroxyphenylacetic acid and homovallinic acid, and norepinephrine. All patients had HF of ischemic etiology. No statistically significant differences were found between the groups with respect to urine volume (1.79 +/- 0.23 L x d( 1) vs 2.20 +/- 0.18 L x d(-1), P =.18) and urinary sodium (161.3 +/- 27.5 mmol x d(-1) vs 232.9 +/- 28.8 mmol x d(-1), P =.12). Urinary L-DOPA was significantly lower in patients with decompensated class III/IV HF than in the other group (79.0 +/- 13.8 nmol x g UCr(-1) vs 108.4 +/- 10.3 nmol x g UCr(-1), P =.04). Urinary dopamine showed a nonstatistically significant trend to be slightly higher (1294.3 +/- 188.5 nmol x g UCr(-1) vs 953.2 +/- 107.4 nmol x g UCr(-1), P =.14). Consequently, urinary dopamine/L-DOPA ratios were markedly higher in patients with decompensated class III/IV HF than in the other patients (20.6 +/- 3.4 vs 9.0 +/- 0.9, P <.001). Plasma L-DOPA (38.1 +/- 4.4 pmol x mL(-1) vs 40.0 +/- 3.0 pmol x mL(-1), P =.48), dopamine (37.0 +/- 6.3 pmol x mL(-1) vs 41.1 +/- 2.6 pmol x mL(-1), P =.53), 3,4-dihydroxyphenylacetic acid (51.7 +/- 11.7 pmol x mL(-1) vs 56.5 +/- 5.4 pmol x mL(-1), P =.09), and norepinephrine (9.5 +/- 2.4 pmol x mL(-1) vs 5.6 +/- 1.0 pmol x mL(-1), P =.12) did not differ between groups. Plasma aldosterone (180.2 +/- 28.0 pg x mL(-1) vs 69.9 +/- 13.3 pg x mL( 1), P <.001) and BNP (677.5 +/- 133.9 pg x mL(-1) vs 389.4 +/- 88.4 pg x mL(-1), P <.04) levels were higher in the decompensated class III/IV HF group than in the other group, whereas serum sodium was lower (137.3 +/- 1.2 mmol x L(-1) vs 143.2 +/- 1.0 mmol x L(-1), P =.001). CONCLUSIONS: These results suggest that, in patients with HF, the increased renal utilization of L-DOPA may constitute a compensatory mechanism, activated in response to stimuli leading to sodium reabsorption. PMID- 11868043 TI - Left bundle-branch block is associated with increased 1-year sudden and total mortality rate in 5517 outpatients with congestive heart failure: a report from the Italian network on congestive heart failure. AB - BACKGROUND: A deleterious effect of complete left bundle-branch block (LBBB) on left ventricular function has been established. Nevertheless, the independent effect of a widened QRS on mortality rate in congestive heart failure (CHF) is still controversial. Therefore, we carried out this analysis to determine whether LBBB is an independent predictor of mortality in CHF. METHODS AND RESULTS: We analyzed the large Italian Network on CHF Registry of unselected outpatients with CHF of different causes. The registry was established by the Italian Association of Hospital Cardiologists in 1995. Complete 1-year follow-up data were available for 5517 patients. The main underlying cardiac diagnosis was ischemic heart disease in 2512 patients (45.6%), dilated cardiomyopathy in 1988 patients (36.0%), and hypertensive heart disease in 714 patients (12.9%). Other causes were recorded in the remaining 303 cases (5.5%). LBBB was present in 1391 patients (25.2%) and was associated with an increased 1-year mortality rate from any cause (hazard ratio, 1.70; 95% confidence interval, 1.41 to 2.05) and sudden death (hazard ratio, 1.58; 95% confidence interval, 1.21 to 2.06). Multivariate analysis showed that such an increased risk was still significant after adjusting for age, underlying cardiac disease, indicators of CHF severity, and prescription of angiotensin-converting enzyme inhibitors and beta-blockers. CONCLUSION: LBBB is an unfavorable prognostic marker in patients with CHF. The negative effect is independent of age, CHF severity, and drug prescriptions. These data may support the rationale of randomized trials to verify the effects on mortality rate of ventricular resynchronization with multisite cardiac pacing in patients with CHF and LBBB. PMID- 11868044 TI - Stability of B-type natriuretic peptide levels during exercise in patients with congestive heart failure: implications for outpatient monitoring with B-type natriuretic peptide. AB - BACKGROUND: B-natriuretic peptide (BNP), a neurohormone secreted from the cardiac ventricles, reflects left ventricular pressure and correlates to disease severity and prognosis. The fact that BNP levels can now be measured by a rapid assay suggests its potential usefulness in the outpatient clinic. However, if patient activity were to markedly alter BNP levels, its use would be less attractive for monitoring patients in the outpatient clinical setting. METHODS: A total of 30 patients (10 normal, 10 New York Heart Association [NYHA] class I-II, 10 NYHA class III-IV) exercised with an upright bicycle protocol. Exercise was carried out to 75% of maximum heart rate, and venous blood was sampled before, immediately after, and 1 hour after completion of exercise. Plasma levels of BNP, epinephrine, and norepinephrine were measured. RESULTS: BNP levels at baseline were 29 +/- 11 pg/mL for normal subjects, 126 +/- 26 pg/mL for NYHA I-II subjects, and 1712 +/- 356 pg/mL for NYHA III-IV subjects. The change in BNP levels with exercise was significantly lower than the change in epinephrine and norepinephrine (P <.001). In normal subjects, BNP increased from 29 pg/mL to 44 pg/mL with peak exercise, still within the range of normal (<100 pg/mL). This is compared with larger increases of norepinephrine (716 pg/mL to 1278 pg/mL) and epinephrine (52 pg/mL to 86 pg/mL) with exercise in normal subjects. There were also only small increases in BNP with exercise in patients with congestive heart failure (NYHA I-II, 30%; NYHA III-IV, 18%). For the same groups, epinephrine levels increased by 218% and 312%, respectively, and norepinephrine levels increased by 232% and 163%, respectively. One hour after completion of exercise, there were only minimal changes in BNP levels from baseline state in normal subjects (+0.9%) and patients with NYHA I-II (3.8%). In patients with NYHA III IV, there was a 15% increase from baseline 1 hour after exercise. CONCLUSIONS: BNP levels show only minor changes with vigorous exercise, making it unlikely that a normal patient would be classified as having congestive heart failure based on a BNP level obtained after activity. Prior activity should not influence BNP levels in patients with congestive heart failure. Therefore, when a patient presents to clinic with a marked change in their BNP level, it may reflect a real change in their condition. PMID- 11868045 TI - Spectrum of heart failure in older patients: results from the National Heart Failure project. AB - BACKGROUND: The elderly make up the majority of patients with heart failure (HF), but information on this segment of the HF population is lacking because clinical trials typically enroll younger patients and population-based studies lack clinical detail. We sought to describe a contemporary national sample of elderly patients with HF and to examine the sample for age-related trends in clinical characteristics. METHODS: We studied the charts of 800 Medicare patients per state who were hospitalized with a principal diagnosis of HF between April 1998 and March 1999. There were 34,587 patients in the sample after exclusion of patients who were <65 years old, repeat discharges, discharges to another acute care facility or against medical advice, or receiving long-term hemodialysis. RESULTS: Comorbidity was common. About one third of patients had chronic obstructive pulmonary disease, about 40% had diabetes, more than half had coronary heart disease, and more than half had a history of hypertension, but comorbidity rates declined with age. Left ventricular ejection fraction was <40% in only 50.4% of patients in whom it was assessed. Associated laboratory abnormalities were relatively constant across the age spectrum, but renal insufficiency was more common with advancing age. The likelihood that patients were in long-term care facilities before admission rose quite steeply with age. CONCLUSIONS: Elderly patients with HF are a heterogeneous group and appear to differ substantially from patients enrolled in clinical trials. Evidence-based guidance for treatment in the context of multiple comorbid conditions, poor renal function, HF with preserved left ventricular systolic function, and residence in long-term care facilities is urgently needed. PMID- 11868046 TI - Ventilatory response to exercise improves risk stratification in patients with chronic heart failure and intermediate functional capacity. AB - BACKGROUND: Peak oxygen consumption (VO2) has an important prognostic role in chronic heart failure (CHF), but its discriminatory power is limited in patients with intermediate exercise capacity (peak VO2 between 10-18 mL/kg/min). Thus, supplementary exertional indexes are greatly needed. METHODS: Six hundred patients with CHF with left ventricular ejection fraction (LVEF) < or = 40% who performed a symptom-limited cardiopulmonary exercise testing were screened and followed up for 780 +/- 450 days. RESULTS: Eighty-seven patients had major cardiac events (77 cardiac deaths and 10 urgent heart transplantations). Multivariate analysis revealed the rate of increase of minute ventilation per unit of increase of carbon dioxide production (VE/VCO2 slope) (chi2, 79.3, P <.0001), LVEF (chi2, 24.6, P <.0001), and peak VO2 (chi2, 9.4, P <.0001) as independent and additional predictors of major cardiac events. VE/VCO2 slope was the strongest independent predictor of outcome (chi2, 20.9, P =.0001) in patients with intermediate peak VO2 (n = 403), and the best cutoff value was 35 (chi2, 25.8; relative risk = 3.2, 95% CI 2.0-5.1, P <.0001). Total mortality rate was 30% in patients with VE/VCO2 slope > or = 35 (n = 103, 26%) and 10% in those with VE/VCO2 slope <35 (n = 300, 74%) (P <.0001). Patients with VE/VCO2 slope > or = 35 had a similar total mortality rate to those with peak VO2 < or = 10 mL/kg/min (30% vs 37%, P not significant). CONCLUSIONS: A rational and pragmatic risk stratification process with symptom-limited cardiopulmonary exercise testing in CHF should include both peak VO2 and VE/VCO2 slope, the latter index effectively predicting outcome in almost one fourth of patients with intermediate exercise capacity. PMID- 11868047 TI - Comparison of ventilatory expired gas parameters used to predict hospitalization in patients with heart failure. AB - BACKGROUND: Several ventilatory expired gas measures obtained during exercise testing demonstrate prognostic value in the heart failure (HF) population. Comparison of prognostic efficacy between pertinent measures is sparse. METHODS: The ability of various expressions of peak oxygen consumption (VO2), the relationship between minute ventilation (VE) and carbon dioxide production (VCO2), and the partial pressure of end-tidal carbon dioxide (P(ET)CO2) were assessed to determine which measure(s) best predicted cardiac-related hospitalization over a 1-year period in subjects diagnosed with HF. RESULTS: Univariate Cox regression analysis found that several expressions of peak VO 2, VE-VCO2 relationship, and P(ET)CO2 were significant predictors of hospitalization. Multivariate Cox regression analysis revealed that the VE/VCO2 slope significantly predicted hospitalization (chi2 = 29.1, P <.00001). Peak VO 2 and P(ET)CO2 did not provide additional predictive value. CONCLUSIONS: The prognostic superiority of the VE/VCO2 slope over peak VO2 may be a result of the latter measure's partial dependence on subject effort and skeletal muscle function. PMID- 11868048 TI - Losartan treatment and left ventricular filling during volume loading in patients with dilated cardiomyopathy. AB - BACKGROUND: Patients with mild heart failure show a reduction in preload reserve mechanism during volume expansion. At this time, the effects of volume expansion on left ventricular (LV) diastolic filling in this subset of patients have not been well characterized. METHODS: We evaluated the effects of acute volume loading on Doppler parameters of LV filling in 10 healthy control subjects and in 12 patients with idiopathic dilated cardiomyopathy (DCM). In patients with DCM, the effects of losartan on diastolic adaptation to volume load were also investigated. RESULTS: During volume loading, the healthy control subjects showed a decrease in isovolumic relaxation time (F = 5.3, P <.05) but an increase in the LV peak filling rate (F = 52.9, P <.001) and velocity time integral of both systolic (F = 72.8, P <.001) and diastolic (F = 4.6, P <.05) pulmonary venous flow. In patients with DCM, isovolumic relaxation time decreased more than in control subjects (F = 8.1, P <.01), and the deceleration time of the early mitral wave was reduced (F = 26.3, P <.001). Furthermore, the duration of pulmonary venous flow reversal exceeded that of mitral flow at atrial contraction (F = 28.5, P <.001). After treatment with losartan, the deceleration time of early mitral wave remained unchanged, and the duration of pulmonary venous flow reversal at atrial contraction did not exceed that of mitral flow; thus, a significant treatment effect was detectable (F = 5.6, P <.05; and F = 6.6, P <.05, respectively). CONCLUSIONS: Control subjects respond to volume load with enhancement in early LV filling, whereas patients with DCM show an increase of LV filling pressure. Diastolic adaptation to volume load improves in patients with DCM after treatment with losartan. PMID- 11868049 TI - A noninvasive measure of baroreflex sensitivity without blood pressure measurement. AB - BACKGROUND: Baroreflex sensitivity (BRS) and heart rate variability (HRV) are attenuated in cardiovascular disease and can give important prognostic information. Conventional measures of BRS require expensive or invasive equipment for the beat-to-beat measure of blood pressure (BP). We examined the possibility of developing a simple protocol that would provide a relatively standardized BP stimulus, which might obviate the need to measure BP beat-by-beat. METHODS AND RESULTS: Fifty-five patients with chronic heart failure (mean age 59 [SD 11] years) and 20 healthy control subjects (mean age 53 [SD 14] years, P not significant) underwent 5-minute recordings of BP (by photoplethysmograph) and R-R interval during 0.1-Hz controlled breathing. The size of the oscillations in BP was the same in the 2 groups (3.6 mm Hg vs 4.1 mm Hg, P =.5). There was, however, a significant difference in the amplitude of the R-R interval oscillations (77 ms vs 31 ms, P <.0001). The amplitude of the R-R interval oscillations correlated strongly with BRS (r = 0.81, P <.0001 with controlled breathing BRS, and r = 0.51, P <.0001 with alpha index). There was no correlation with the size of BP oscillations (r = -0.13, P not significant with controlled breathing BRS, and r = -0.15, P not significant with alpha index). In a separate study, a group of 22 young patients (mean age 36 years) with type I diabetes mellitus and 28 healthy control subjects (mean age 39 years) underwent measurement of resting HRV and amplitude of R-R interval oscillations during 0.1-Hz breathing. There was no significant difference in triangular index or low-frequency R-R interval power between the 2 groups. There was, however, a significant difference in the amplitude of R-R interval oscillations during controlled breathing between patients with diabetes and healthy control subjects. Total and high-frequency RR interval variability was also significantly different between the 2 groups. CONCLUSION: During 0.1-Hz breathing, the marked difference in BRS between patients with CHF and age-matched control subjects is the result of smaller R-R interval oscillations. In young patients with diabetes, these R-R interval oscillations are significantly smaller than age-matched control subjects, even when some measures of spontaneous HRV are not different between groups. Breathing at 0.1 Hz provides a standard BP stimulus and concentrates spectral power of heart rate at one frequency, enabling simple evaluation of BRS even when BP measurement is not available. PMID- 11868050 TI - Tako-tsubo-like left ventricular dysfunction with ST-segment elevation: a novel cardiac syndrome mimicking acute myocardial infarction. AB - BACKGROUND: Peculiar asynergy, which consists of hypokinesis or akinesis from the mid portion to the apical area and hyperkinesis of the basal area on contrast left ventriculogram, is rare. Because the end-systolic left ventriculogram looks like a "tako-tsubo," which was used for trapping octopuses in Japan, we proposed the term "tako-tsubo-like left ventricular dysfunction." Our aim was to evaluate its clinical features and causes. METHODS: We studied 30 patients with tako-tsubo like left ventricular dysfunction without significant coronary artery disease. We assessed its pathophysiologic mechanisms by coronary spasm provocation test, endomyocardial biopsy, measurement of virus titer, and measurement of circulating catecholamine levels. RESULTS: Patient age ranged from 55 to 83 years. Twenty eight were women and 2 were men. Tako-tsubo-like left ventricular dysfunction was dramatically resolved on predischarge left ventriculogram at 11.3 +/- 4.3 days. Acute coronary angiography revealed spontaneous multivessel coronary spasm in 3 patients. Among 14 patients, ergonovine or acetylcholine induced epicardial single coronary spasm in 4 patients and multivessel coronary spasm in 6 patients. Spontaneous microvascular spasm occurred at predischarge in 1 patient. An endomyocardial biopsy specimen in 3 patients and measurement of virus titer in 7 patients did not show evidence of acute myocarditis. Circulating norepinephrine was normal or slightly elevated in 6 patients. CONCLUSIONS: We showed clinical features of a novel cardiac syndrome with tako-tsubo-like left ventricular dysfunction. Although the precise cause remains unclear, simultaneous multivessel coronary spasm at the epicardial artery or microvascular levels may contribute to the onset of tako-tsubo-like left ventricular dysfunction. PMID- 11868051 TI - Coronary calcium and cardiovascular event risk: evaluation by age- and sex specific quartiles. AB - BACKGROUND: Wide differences in risks for cardiovascular disease (CVD) events associated with coronary artery calcium (CAC) have been reported. We evaluated the relationship of the coronary calcium quartile, on the basis of age-sex cut points from a large sample of asymptomatic patients, to CVD events as a possible standardized means for reporting event risks associated with CAC, in comparison with quartiles of absolute CAC scores. METHODS: We applied age/sex-stratified cut points to 928 asymptomatic men and women (mean age 54 years) followed up for an average of 3.3 years, during which 28 CVD events were confirmed. Cox regression was used to evaluate the relation of the 2nd, 3rd, and 4th quartiles (compared with the 1st), with and without age/sex stratification, to the risk of future CVD events. RESULTS: The number of events (and percent incidence) that occurred in the 1st, 2nd, 3rd, and 4th quartiles of coronary calcium was 4 (0.9%), 2 (2.0%), 9 (4.5%), and 13 (6.4%) events (P =.001) for the age/sex-stratified quartiles and 4 (1.0%), 0 (0%), 7 (3.0%), and 17 (7.3%) for the absolute score quartiles (P =.001). In multivariable analysis adjusted for other risk factors, there was a modest increase in CVD events seen among those in the 3rd quartile (relative risk [RR] 4.3, P =.02), with a greater risk seen among those in the 4th quartile (RR 6.0, P <.01) (compared with the 1st quartile). This did not differ from use of absolute CAC scores, where RR = 2.6 (P =.14) for the 3rd quartile and RR = 6.4 (P <.01) for the 4th quartile. CONCLUSIONS: Our results suggest that age-sex stratification by percentile rank of CAC is as accurate as absolute CAC scores for predicting CVD events in asymptomatic persons. Ongoing longitudinal population-based studies will provide more definitive data. PMID- 11868052 TI - Association of anxiety with reduced baroreflex cardiac control in patients after acute myocardial infarction. AB - BACKGROUND: Although depression has been associated with increased mortality in patients after acute myocardial infarction (AMI), little is known about the effects of depression on autonomic nervous system control of heart rate. This study evaluated whether depression is associated with impaired baroreflex sensitivity (BRS) in patients with AMI. METHODS: Two hundred four hospitalized patients with AMI were evaluated 6 +/- 3 (mean +/- SD) days after AMI. BRS was assessed using cross-spectral analysis to measure baroreceptor-mediated R-R interval oscillations. Depression was determined using the Diagnostic Interview Schedule, and severity of depressive symptoms was measured with the Beck Depression Inventory. In order to adjust for possible differences in anxiety, we also measured state anxiety using the Spielberger State Anxiety Inventory. RESULTS: Depression was not significantly related to BRS. However, anxiety was significantly related to low BRS in multivariate analysis, after the potentially confounding variables of age, blood pressure, and respiratory frequency were controlled for. Comparison of groups with high and low anxiety (on the basis of a median split of state anxiety scores) showed that BRS was reduced by approximately 20% in the patients with the higher anxiety scores (4.7 +/- 3.2 ms/mm Hg vs 5.7 +/- 3.3 ms/mm Hg, P <.05), after adjustment for differences in age, blood pressure, and respiratory frequency. CONCLUSIONS: High levels of anxiety, but not depression, are associated with reduced vagal control in patients after AMI. PMID- 11868053 TI - Inverse association between carotid intima-media thickness and the antioxidant lycopene in atherosclerosis. AB - BACKGROUND: Antioxidants may prevent atherosclerosis by interfering with endothelial activation, which involves the expression of endothelial adhesion molecules. The aim of this study was to explore the relationship between plasma levels of some lipid-soluble antioxidants (gamma-tocopherol, alpha-tocopherol, lycopene, beta-carotene, and ubiquinone), carotid maximum intima-media thickness (IMTmax), an index of atherosclerotic extension/severity, and soluble adhesion molecules (vascular cell adhesion molecule-1 [VCAM-1], intercellular adhesion molecule-1 [ICAM-1], and E-selectin), which are taken as a reflection of vascular cell expression of adhesion molecules. METHODS: We studied 11 healthy control subjects, 11 patients with uncomplicated hypertension (UH), and 11 patients with essential hypertension plus peripheral vascular disease (PVD) who were matched for age, sex, smoking habit, and body mass index. RESULTS: Patients with PVD had elevated IMTmax (2.7 [1.1-3.1] mm, median [range]) compared with both patients with UH(1.2 [0.8-2.4] mm) and control subjects (1.0 [0.6-2] mm). In patients with PVD, soluble (s)VCAM-1 and sICAM-1 were also significantly higher than in the 2 other categories. Plasma levels of lycopene had a trend toward lower values in patients with PVD compared with other groups (P =.13). A statistically significant correlation was found between lycopene and IMTmax (r = 0.42, P =.014) at univariate analysis, which persisted at multivariate analysis (P <.05) and was independent of low-density lipoprotein cholesterol, creatinine clearance, and plasma insulin. Plasma lycopene did not significantly correlate with any of the soluble adhesion molecules tested. CONCLUSIONS: We conclude that the inverse relationship of plasma lycopene with IMTmax is compatible with a protective role of this natural dietary antioxidant in atherosclerosis, although the mechanism of protection does not apparently involve a decrease in endothelial activation measured through soluble adhesion molecules. PMID- 11868054 TI - Nonsteroidal antiinflammatory drugs after acute myocardial infarction. AB - BACKGROUND: Many older patients undergo therapy with nonsteroidal antiinflammatory drugs (NSAIDs), medications that produce effects on platelets and inflammation similar to those produced with aspirin. The impact of these agents on mortality after acute myocardial infarction is not known. We sought to determine whether the use of NSAIDs alone, or in addition to aspirin, is associated with a lower 1-year mortality rate in elderly patients after acute myocardial infarction. METHODS: We performed an analysis of the Cooperative Cardiovascular Project, a retrospective medical record review that included demographic and clinical information for Medicare beneficiaries hospitalized with a diagnosis of acute myocardial infarction during 1994 and 1995. The cohort included 48,584 elderly patients with acute myocardial infarction without contraindications to NSAID or aspirin therapy. There were 736 patients (1.5%) who were prescribed NSAIDs alone, 36,211 (74.5%) prescribed aspirin alone, 2096 (4.3%) prescribed both NSAIDs and aspirin, and 9541 (19.6%) prescribed neither medication at discharge. RESULTS: Compared with patients discharged with neither medication, prescriptions of NSAID therapy alone (hazard ratio [HR], 0.77; 95% CI, 0.65-0.90), aspirin alone (HR, 0.81; 95% CI, 0.77-0.86), and both medications (HR, 0.78; 95% CI, 0.69-0.88) were associated with lower adjusted 1-year mortality rates. Compared with patients prescribed NSAID therapy at discharge, there was no significant benefit associated with the addition of aspirin, and the benefit of NSAID therapy was not significantly different from that of aspirin alone. CONCLUSION: The prescription of NSAID therapy at hospital discharge for elderly Medicare beneficiaries who survived acute myocardial infarction was associated with similarly lower 1-year mortality rates as compared with aspirin therapy. The addition of aspirin to NSAID therapy was not associated with an additional survival benefit. PMID- 11868055 TI - Slow upsloping ST-segment depression during exercise: does it really signify a positive stress test? AB - BACKGROUND: Slow upsloping ST-segment depression during stress is thought to represent an ischemic response to exercise treadmill testing (ETT). AIM: We used modern single-photon emission computed tomography (SPECT) imaging protocols to determine the incidence of ischemia in patients with slow upsloping ST depression during exercise and whether this response signifies more or less severe coronary artery disease (CAD) and risk in comparison with rapid upsloping ST depression and particularly with horizontal or downsloping ST depression. METHODS: We enrolled 33 patients (group 1) with rapid upsloping ST depression (>1 mm extending <0.08 seconds beyond J point), 32 patients (group 2) with slow upsloping depression (>1.5 mm extending >0.08 seconds beyond J point), and 35 patients (group 3) with horizontal or downsloping depression (>1 mm at 0.08 seconds beyond J point). Summed stress score (SSS), summed difference score (SDS), stress extent percent (SE%) and reversible extent percent (RE%) of perfusion abnormalities, lung-heart ratio (LHR), and transient ischemic dilatation (TID) were calculated. RESULTS: The mean SSS, SDS, SE%, RE%, and LHR were similar between groups 1 and 2 but significantly higher in group 3. Incidence of ischemia was similar in groups 1 and 2 (39% and 25%) but significantly higher in group 3 (77%, P <.001). Evidence of TID was seen in none of the patients in groups 1, in 3% of patients in group 2, and in 23% of patients in group 3. CONCLUSIONS: Slow upsloping ST depression does not signify more severe ischemia, more extensive CAD, or more stress-induced backward left ventricular failure. Thus, it would be reasonable to consider patients with slow upsloping ST depression during exercise as having a very low likelihood of CAD, similar to patients with rapid upsloping ST depression. PMID- 11868056 TI - Is there a benefit to early angiography in patients with ST-segment depression myocardial infarction? An observational study. AB - BACKGROUND AND OBJECTIVE: Patients with ST-segment depression myocardial infarction (MI) have a similar long-term outcome compared with those who have ST segment elevation. It remains unclear whether an aggressive treatment approach with very early (<6 hours) angiography and revascularization improves outcome over an early conservative approach. We compared the short-term outcome of patients who received very early (<6 hours) angiography with patients who received early conservative therapy for ST-segment depression MI. METHODS: Patients seen within 12 hours with ST-segment depression on the initial electrocardiogram (ECG) were identified from the National Registry of Myocardial Infarction 2 (NRMI) database, which collected information from 1994 to 1998. Those who received very early (<6 hours) angiography were compared with those who received early conservative therapy. The short-term outcomes, including major bleeding episodes, cerebral vascular events, recurrent ischemia and angina, MI, and death, were compared on the basis of the initial therapy received. RESULTS: Patients treated with very early angiography (2402) were younger, more likely to be males, smokers, and have less of a cardiac history (angina, MI, congestive heart failure, aortocoronary bypass surgery) and less likely to have diabetes mellitus than those who received early conservative therapy (17,735). Hospital outcome in the very early angiography group was similar to the early conservative therapy group in terms of cerebral vascular events (0.8% vs 1.0%, P =.27), major bleeding events (2.8% vs 2.4%, P =.25), and recurrent MI (2.1% vs 2.3%, P =.57) but was associated with lower recurrent ischemia or angina (11.4% vs 16.7%, P <.001) and improved survival (death, 4.9% vs 7.3%, P <.001). Multivariate analysis suggested that patients receiving very early angiography had lower mortality compared with those receiving early conservative therapy (odds ratio [OR] = 0.76; 95% CI 0.60-0.95). However, comparing patients matched on propensity score (1405) showed mortality was similar in both treatment groups (5.6% vs 5.4%, P =.87), with no significant inhospital mortality benefit of very early angiography (OR = 0.89; 95% CI 0.71-1.13). CONCLUSIONS: The apparent mortality benefit of very early angiography in patients with ST-segment depression MI is a reflection of bias by confounding. Controlling for baseline differences using propensity score methods in this observational study indicated no inhospital mortality benefit of a very early aggressive approach compared with a conservative approach. PMID- 11868057 TI - Long-term outcome of primary percutaneous transluminal coronary angioplasty for low-risk acute myocardial infarction in patients older than 80 years: a single center, open, randomized trial. AB - BACKGROUND: Although coronary reperfusion therapy with thrombolytic agents or percutaneous transluminal coronary angioplasty (PTCA) immediately after acute myocardial infarction (AMI) has survival benefits for younger patients, the effect of coronary reperfusion therapy for very elderly (aged 80 years and older) patients with AMI remains controversial. METHODS AND RESULTS: We studied 120 patients aged 80 years and older at relatively low risk with AMI. The patients were randomized into a primary PTCA group (n = 61) or a "conservative" no-PTCA group (n = 59). Long-term follow-up examination was conducted with regard to endpoints, which included all causes of death, cardiac death, nonfatal re-MI, the development of congestive heart failure, and cerebral vascular accident. End diastolic volume index and end-systolic volume index were significantly increased in both groups at follow-up examination 6 months after AMI. However, left ventricular ejection fraction, end-diastolic volume index, and end-systolic volume index were similar between both groups. With endpoints of all causes of death, cardiac death, reinfarction, congestive heart failure, and cerebral vascular accident, a 3-year Kaplan-Meier event-free survival rate analysis revealed no significant benefits in the PTCA group. Anteroseptal MI, multivessel disease, and left ventricular ejection fraction were significantly associated with the combined events with multivariate Cox proportional hazards analysis results. CONCLUSION: First, primary PTCA for very elderly patients with AMI appears to have few beneficial effects on combined events during a 3-year period. Second, early PTCA did not prevent left ventricle remodeling after AMI in patients with AMI at relatively low risk. PMID- 11868058 TI - The impact of tranilast on restenosis after coronary angioplasty: the Second Tranilast Restenosis Following Angioplasty Trial (TREAT-2). AB - BACKGROUND: The Tranilast Restenosis Following Angioplasty Trial showed that oral administration of 600 mg/day of tranilast for 3 months markedly reduced the restenosis rate after percutaneous transluminal coronary angioplasty (PTCA) for de novo lesions. METHODS: We conducted the second multicenter, randomized, double blinded placebo-controlled trial. A total of 297 patients with 329 lesions were randomly assigned to treatment with tranilast or a placebo for 3 months after successful PTCA for both de novo and restenotic lesions. Angiographic follow-up examination was done at 3 months, and angiograms were interpreted with a quantitative approach. RESULTS: Two hundred thirty-nine lesions (72.6%) in 216 of the patients (72.7%) met the criteria and were included in the assessment of restenosis. Lesion restenosis was defined as a loss of 50% or more of the initial gain, and the restenosis rates were 18.8% in the tranilast group (n = 112) and 44.1% in the placebo group (n = 127; P =.00005). The restenosis rate, defined as a percent stenosis of > or = 50% at follow-up examination, was also significantly lower in the tranilast group (25.9% versus 41.9%; P =.012). The numbers of restenotic lesions were 38 (33.9% of 112) in the tranilast group and 30 (23.6% of 127) in the placebo group. In restenotic lesions, the lesion restenosis rate was significantly lower in the tranilast subgroup (18.4% versus 53.3% with the first restenosis criterion; P =.004). CONCLUSION: The oral administration of tranilast for 3 months markedly reduced the restenosis rate after PTCA, even in restenotic lesions. PMID- 11868059 TI - Effect of very-low-dose niacin on high-density lipoprotein in patients undergoing long-term statin therapy. AB - BACKGROUND: A low level of high-density lipoprotein (HDLC) is a proven risk factor for coronary artery disease. Niacin raises HDLC levels, but it is infrequently used because of its side effect profile. Niacin's side effects are dose related. This study tests the hypothesis that very low-dose niacin, in conjunction with long-term statin therapy, will improve the lipid profile by significantly raising the level of HDLC, with fewer side effects than traditional doses of niacin. METHODS: Fifty patients undergoing stable statin therapy for 3 months were blindly randomized to receive either placebo or niacin 50 mg administered by mouth 2 times daily for 3 months. Patients with diabetes and active smokers were excluded. Each patient completed a questionnaire regarding current medical problems, medications, and lifestyle before and after the therapy. Patients were questioned about any possible side effects that occurred during the medication trial. The primary end points were change in HDLC level and patient-reported side effects. RESULTS: Thirty-nine patients completed the study. Very low-dose niacin added to statin therapy increased the mean HDLC, 2.1 mg/dL in niacin group (standard error of the mean, 0.767) versus -0.56 mg/dL for placebo group (standard error of the mean-.816, P =.0246 by analysis of variance). Five patients receiving niacin, versus 2 patients receiving placebo, had episodes of flushing. No major side effects were noted. No patients stopped the study medication as a result of side effects. CONCLUSIONS: The addition of very low-dose niacin to statin therapy increased HDLC cholesterol significantly, while avoiding the side effects that are associated with traditional doses of niacin therapy. PMID- 11868060 TI - Recent trends in the incidence rates of and death rates from atrial fibrillation complicating initial acute myocardial infarction: a community-wide perspective. AB - BACKGROUND: Limited information is available about recent trends in the incidence and death rates from atrial fibrillation (AF) complicating acute myocardial infarction (AMI). The purpose of this study was to examine the impact and trends over time of AF complicating initial AMI. METHODS: We conducted a longitudinal study of 2596 residents of the Worcester, Massachusetts, metropolitan area with initial AMI and without previous AF who were hospitalized at all area hospitals in 5 annual periods between 1990 and 1997. RESULTS: A total of 13% of hospitalized patients developed AF. There was a marked decrease in the proportion of patients who developed AF over time (18% in 1990; 11% in 1997). Patients hospitalized during the most recent study years remained at significantly lower risk for developing AF than patients hospitalized in 1990 after controlling for factors that might affect the risk of AF. Patients with AF were more likely to die during hospitalization than those without AF even after controlling for other prognostic factors. Despite the adverse impact of AF, we observed trends toward improved hospital survival during the most recent study years in patients developing AF. Patients discharged after developing AF had higher long-term death rates than patients who did not develop AF, although these differences were attenuated after adjusting for other factors. CONCLUSIONS: The results of this community-wide study suggest a significant decline in the incidence of AF complicating AMI. Although patients with AF had higher hospital death rates, we observed trends of improved survival for these patients. AF remains a common complication of AMI to which targeted treatment efforts appear warranted. PMID- 11868061 TI - Beta blockers normalize QT hysteresis in long QT syndrome. AB - OBJECTIVES: This study was performed to evaluate the impact of beta blockers on QT adaptation to heart rate during the exercise and recovery phases of exercise testing in long QT syndrome. BACKGROUND: Long QT syndrome is characterized by familial syncope and sudden death in the context of sudden heart rate changes. QT hysteresis has been proposed as a phenotypic marker of long QT syndrome, suggesting altered QT adaptation to changes in heart rate. METHODS: Fourteen patients with long QT syndrome (aged 26 +/- 16 years, 6 male) and 10 healthy volunteers (aged 37 +/- 11 years, 9 male) underwent graded exercise testing with continuous lead II electrocardiographic monitoring. Long QT patients underwent repeat assessment after 1 month of beta blockade. QT intervals at matching heart rates were compared during exercise and recovery to determine the effect of beta blockade on QT hysteresis, defined as the recovery QT peak interval subtracted from the exercise QT peak interval. RESULTS: In the 14 long QT syndrome patients, beta blockers slowed the resting heart rate without affecting the corrected QT interval (502 +/- 52 ms baseline vs 481 +/- 40 ms beta blocker, P =.17). The increase in heart rate with exercise was similar in the 3 groups (P =.73). Exaggerated hysteresis of the QT interval was seen in the patients with long QT syndrome at baseline compared with controls (46 +/-19 ms vs 19 +/- 11 ms 1 minute into recovery, P =.006). Beta blockers had minimal effect on the QT interval but markedly reduced hysteresis with minimal separation of the exercise and recovery QT/RR curves (25 +/- 35 ms 1 minute into recovery, P =.027). The combined curve separation at all 6 time points analyzed was 165 +/- 95 ms in patients with long QT syndrome at baseline, 40 +/- 131 ms after beta blockade, and 29 +/- 30 ms in control subjects (P =.002). Comparison of the beta blocker effect on hysteresis in the 2 genotypes suggested a greater reduction in hysteresis in the 3 patients with long QT syndrome 1 compared with the 11 patients with long QT syndrome 2. CONCLUSIONS: Beta blockers reduce QT hysteresis in patients with long QT syndrome to values seen in normal patients. This improved QT adaptation to changes in heart rate may explain the clinical benefit of beta blockers in long QT syndrome. PMID- 11868062 TI - Prevalence of premature ventricular contractions in a population of African American and white men and women: the Atherosclerosis Risk in Communities (ARIC) study. AB - BACKGROUND: The distribution or the causes of premature ventricular contractions (PVCs) in diverse populations are not fully known. We describe the prevalence of PVCs on a 2-minute electrocardiogram (ECG) in adults to determine whether hypertension has an important association with such PVCs. METHODS: A cross sectional analysis of the 15,792 individuals (aged 45-65 years) from the four US communities participating at visit 1 of the Atherosclerosis Risk In Communities (ARIC) study was performed. Multiple logistic regression was used to determine the association of PVCs with potential causal predictors of PVCs. RESULTS: Based on a 2-minute ECG, PVCs are present in >6% of middle-aged adults. Increasing age, the presence of heart disease, faster sinus rates, African American ethnicity, male sex, lower educational attainment, and lower serum magnesium or potassium levels are directly related to PVC prevalence. Independently of these factors, hypertension is associated with a 23% increase in the prevalence of PVCs. CONCLUSIONS: The prevalence of PVCs on a 2-minute ECG differs by age, ethnicity, and sex and is associated with hypertension, heart disease, faster sinus rates, electrolyte abnormalities, and lower educational attainment. Hypertension is likely to be a major cause of PVCs in adults. PMID- 11868063 TI - Effect of shock polarity on the efficacy of transthoracic atrial defibrillation. AB - BACKGROUND: The energy requirement for internal ventricular defibrillation is reduced by reversal of shock polarity. The influence of shock polarity on the efficacy of transthoracic atrial defibrillation is unknown. METHODS: This prospective, randomized study enrolled 110 consecutive patients who were referred for elective cardioversion of persistent atrial fibrillation (AF). The electrodes were placed in the anteroposterior position. The patients were randomized to receive either standard (anterior pad = cathode) or reversed polarity (anterior pad = anode) shocks with a damped sinusoidal monophasic waveform. A step-up protocol was used to estimate the cardioversion threshold. The initial shock energy was 50 J, with subsequent increments to 100, 200, 300, and 360 J in the event of cardioversion failure. RESULTS: Sixty-four percent of the patient population were men, with a mean age of 66 +/- 13 years and a mean duration of AF of 242 +/- 556 days. The overall success rates of cardioversion were 84% for standard polarity and 78% for reversed polarity (P not significant). Among the patients who were successfully cardioverted, the mean atrial defibrillation threshold was 198 +/- 103 J for standard polarity and 212 +/- 107 J for reversed polarity (P not significant). CONCLUSIONS: Reversal of shock polarity does not improve transthoracic cardioversion efficacy with a standard damped sinusoidal monophasic waveform. Alternate strategies should be considered for patients who fail external cardioversion, such as adjunctive pharmacologic treatment, use of a biphasic shock waveform, or internal cardioversion. PMID- 11868065 TI - Early systolic mitral annular motion velocities responses to dobutamine infusion predict myocardial viability in patients with previous myocardial infarction. AB - OBJECTIVE: This study was undertaken for the determination of the correlation between myocardial viability and regional systolic mitral annular motion velocity (MAV) response to dobutamine stress in patients with previous myocardial infarction (MI) with pulsed tissue Doppler scan imaging. METHODS: The study included 45 patients (mean age, 65 +/- 12 years) with previous MI with 1 major coronary lesion and 30 healthy individuals (mean age, 61 +/- 14 years). 99mTc methoxyisobutylisonitrile scintigraphy was performed to divide the patients into 2 groups: the viability (+) group (n = 25) and the viability (-) group (n = 20). Dobutamine was infused (at 2, 5, 10, and 20 microg/kg/min), and the peak first and second systolic MAVs (Sw1 and Sw2, respectively) were measured at the level of the mitral annulus corresponding to the infarct regions in the MI group and to the 6 mitral annular sites in the control group. In addition, the left ventricular wall motion score index (WMSI) was determined with 2-dimensional echocardiography. RESULTS: At baseline, the WMSI was significantly greater and the mean Sw1 and Sw2 were significantly lower in both the viability (+) and (-) groups than in the control group, but there were no significant differences between the viability (+) and (-) groups. After dobutamine infusion, the WMSI improved only in the viability (+) group. The mean Sw1 and Sw2 increased significantly with 2 microg/kg/min and 5 microg/kg/min of dobutamine, respectively, in the viability (+) group. With an increase in Sw1 of 2.0 cm/s or more with 5 microg/kg/min of dobutamine, viable myocardium was detected, with a sensitivity of 92% and a specificity of 90%. There were no significant increases in Sw1 or Sw2 in the viability (-) group with dobutamine infusion. CONCLUSION: Viable left ventricular myocardium is identified with peak early systolic MAV during dobutamine infusion. PMID- 11868064 TI - Myocardial function and geometry in hypertensive subjects with low levels of afterload. AB - BACKGROUND: It has been hypothesized that the level of end-systolic wall stress (sigma(m)) is a feedback signal that regulates the level of hypertrophy. Thus, low levels of sigma(m) may signify inappropriate hypertrophy. METHODS: To characterize left ventricular (LV) structure and systolic function in hypertensive subjects with low levels of sigma(m), we studied 763 patients. LV function was studied by midwall stress-shortening analysis. Partition values for sigma(m) were derived from a separate group of normal subjects, and the study population was divided into low stress (group I, n = 136), high stress (group III, n = 157), and intermediate stress group II (n = 470). LV chamber and myocardial function were characterized by relating shortening at the endocardium and at the midwall, respectively, to stress. RESULTS: As expected, group III patients had the highest values for systolic blood pressure and LV cavity size but the lowest values for wall thickness and relative wall thickness. Surprisingly, however, there were no significant differences among stress groups with regard to age or body mass index. Contrary to the hypothesis that low levels of stress are indicative of excessive hypertrophy, there were no significant differences among the 3 groups with regard to LV mass or any form of LV mass index. Furthermore, despite lower mean values for afterload, group I patients had significantly lower values for midwall shortening, and this finding was indicative of reduced myocardial function; stress-shortening plots demonstrated that 28% of group I patients fell below 95% CI compared with 10% of group II and only 5% of group III patients. CONCLUSIONS: Hypertensive subjects with low values for sigma(m) have more concentric LV geometry (higher relative wall thickness) and, on average, reduced myocardial function. PMID- 11868066 TI - Imaging of acute abdominal emergencies in infants and children. PMID- 11868069 TI - Laser-induced thermotherapy: do we really need MR thermometry? PMID- 11868070 TI - Advances in ultrasound. AB - Significant advances have been recently introduced into various fields of technology, taking advantage of the use of new piezoelectric materials and the large diffusion of broadband transducers. Various types of modulation may be applied to the pulse characteristics, using single pulse, multipulse or multiline techniques, and resulting in improved spatial resolution and better penetration. Non-linear imaging uses the harmonics component, which is generated by tissues or by contrast agents. Different modalities can be used to separate harmonics from fundamental bands from the received signal. New Doppler modes have been developed, whereas grey-scale flow imaging allows the simultaneous imaging of blood flow and tissues. Compounding techniques improve the contrast resolution of tissues and reduce artefacts. If 3D techniques are now currently available, real time 4D imaging has been recently introduced. Elastographic imaging is still under evaluation, but promising clinical results have been shown. Recent release of the DICOM specification has made the full integration of ultrasound to the PACS systems easier. All these advances indicate that the contribution and potential of ultrasound in patient management is still growing. PMID- 11868071 TI - Scrotal ultrasound. PMID- 11868072 TI - Ultrasound of muscles. AB - Muscles are among the soft tissues one of the best adapted to ultrasound examination. In fact, it was the first imaging available for the evaluation of muscle disease. The availability, low cost, and ease of examination makes ultrasound superior to MRI for follow-up of lesions and searching for healing problems such as as fibrosis, cystic haematomas, or myositis ossificans. When dealing with fresh traumatic muscle lesions, the main goal of ultrasound is to assess the presence of a muscle tear or not. Haematoma is the key sign of a muscle tear. The ideal time for the examination is between 2 and 48 h after the muscle trauma. Before 2 h, the haematoma is still in formation. After 48 h, the haematoma can be spread outside of the muscle. After healing, ultrasound can depict some complications such as a cystic lesion or myositis ossificans. Muscle atrophy, inflammation, avulsion and tumours are also good indications for ultrasound. PMID- 11868073 TI - Ultrasound of tendons and nerves. AB - Tendons and nerves represent probably one of the best application of musculoskeletal US due to the high lesion detection rate and accuracy of US combined with its low cost, wide availability, and ease of use. The refinement of high-frequency broadband linear-array transducers, and sensitive color and power Doppler technology, have improved the ability of US to detect fine textural abnormalities of these structures as well as to identify a variety of pathological conditions. Characteristic echotextural patterns, closely resembling the histological ones, are typically depicted in these structures using high US frequencies. In tendon imaging, US can assess dislocations, degenerative changes and tendon tears, including intrasubstance tears, longitudinal splits, partial and complete rupture, inflammatory conditions and tendon tumors, as well as postoperative findings. In nerve imaging, US can support clinical and electrophysiological testing for detection of compressing lesions caused by nerve entrapment in a variety of osteofibrous tunnels of the limbs and extremities. Congenital anomalies, nerve tears, and neurogenic tumors can also be diagnosed. Overall, US is an effective technique for imaging tendons and nerves. In most cases, a focused US examination can be performed more rapidly and efficiently than MR imaging. PMID- 11868074 TI - Ultrasound of the joints. AB - High-frequency ultrasound is now considered an excellent modality to image normal tendons, muscles, and peripheral nerves as well as to diagnose a wide variety of pathological conditions affecting these structures. Although US is limited in the visualisation of some intra-articular structures, it can be a useful tool in joint disease assessment. Ultrasound has some advantages over other imaging modalities including time- and cost-effectiveness, superior spatial resolution, dynamic examination and the possibility to perform the examination in a comfortable position for the patient. The aims of this review are twofold: firstly, to present the normal US appearance of the joint structures that are susceptible to US examination, including the joint surfaces, intra-articular structures such as menisci and other fibrocartilages, capsule and ligaments; and secondly, to show the US appearance of the most commonly encountered joint disorders and discuss the role of US in the imaging strategy of joint disorders. PMID- 11868075 TI - MRI characteristics in focal hepatic disease before and after administration of MnDPDP: discriminant analysis as a diagnostic tool. AB - The aim of this study was to determine if different types of focal hepatic lesions can be differentiated by specific quantitative and qualitative imaging characteristics pre- and post-Mangafodipir trisodium (MnDPDP) administration using a computerized multivariable, discriminant analysis (DA). In a multicenter trial, 151 patients with focal liver disease were studied at 1.5 and 1.0 T using gradient-recalled echo T1 and fast spin-echo T2-weighted images pre and post MnDPDP (0.005 mmol/kg b.w.) i.v. administration. Analysis could be performed in 141 of 151 of the patients. The variables used in both single variable analysis and DA included contrast-to-noise ratios pre and post MnDPDP, presence of rim enhancement, margin, and heterogeneity of a lesion pre and post MnDPDP. The classification of diagnoses using DA was compared with a standard of reference (HCC in 23%, metastases in 25%, cyst in 13%, FNH in 10%, hemangioma in 11%, and other or no lesion in 18% of the patients; histology in 49%, long-term follow-up in 51% of the cases). In the differentiation of the various hepatic lesions, CNR together with the presence of heterogeneity or rim enhancement as variables for DA gave the highest sensitivity, specificity, and accuracy which ranged between 65 and 93, 44 and 83, and 65 and 86%, respectively. The DA models based on post MnDPDP variables showed better classification results than the models based on pre-MnDPDP variables. An improvement of accuracy was observed when differentiating HCC from FNH lesion groups (48.9-67.4%; p < or = 0.05), and when differentiating HCC from metastasis lesion groups (68.3-84.1%; p < or = 0.01). In all regards there was no difference for T2-weighted images pre and post MnDPDP. By combining quantitative and qualitative variables, DA proved to be a useful tool in lesion discrimination. Due to considerable heterogeneity within some of the lesion type groups, the definite diagnostic impact of MnDPDP cannot be completely established yet, and further investigation is still necessary. PMID- 11868076 TI - Dilatation of the intrahepatic bile ducts associated with benign liver lesions: an unusual finding. AB - In three patients presenting different types of liver lesions, including isolated cyst, focal nodular hyperplasia (FNH), and hemangioma, intrahepatic bile duct dilatation was observed on US and CT. Final diagnosis was obtained by surgery in two cases (cyst and FNH) and by 1-year follow-up in one patient presenting an isolated hemangioma. The only common characteristic in our three cases was that lesions were present in segment four according to Couinaud's classification, at the level of the transverse fissure, suggesting that a space-occupying lesion at this site may cause compression of the common hepatic duct and right or left intrahepatic bile ducts. Our report indicates that compression may occur even with lesion of moderate size (35-40 mm in diameter). A benign liver lesion may cause a bile duct dilatation, particularly if located in segment 4, close to the hilum. Awareness of this possibility is important to avoid unnecessary invasive diagnostic procedures, particularly when all imaging criteria are consistent with a benign lesion. PMID- 11868077 TI - Long-term follow-up after neoplastic seeding complicating percutaneous ethanol injection for treatment of hepatocellular carcinoma. AB - We describe a case of subcutaneous metastasis along the needle track after percutaneous ethanol injection (PEI) for treatment of hepatocellular carcinoma. After surgical resection and extrabeam radiation therapy the patient is alive without evidence of recurrence five years after PEI. One should pay attention to the abdominal wall around the needle track in interpreting CT or MR images of patients with previous PEI. PMID- 11868078 TI - Computer-aided diagnosis in virtual colonography via combination of surface normal and sphere fitting methods. AB - The success of CT colonography (CTC) depends on appropriate tools for quick and accurate diagnostic reading. Current advancements in computer technology have the potential to bring such tools even to personal computer level. In this paper a technique for computed-aided diagnosis (CAD) using CT colonography is described. The method uses a combination of surface normal and sphere fitting methods to label positions in the volume data, which have a strong likelihood of being polyps, and presents them in a user-friendly way. The method was tested on a study group of 18 patients and the detection rate for polyps of 10 mm or larger was 100%, comparable to that of human readers. The price paid for a high detection rate was a large number of approximately eight false-positive findings per case. Our results show that CAD is feasible, and if the number of false positives is further reduced, then this method can be useful for clinical screenings. PMID- 11868079 TI - Giant Meckel's diverticulum containing enteroliths: typical CT imaging findings. AB - We report a case of a giant Meckel's diverticulum containing numerous enteroliths. A correct diagnosis was made preoperatively by means of CT, by demonstrating a connection between the diverticulum, containing multiple peripherally calcified stones, and the small intestine. PMID- 11868080 TI - Cross-sectional imaging of primary osseous hemangiopericytoma. AB - The aim of this study was to assess cross-sectional imaging features and the value of CT and MRI in primary hemangiopericytoma of bone. In five patients with histologically proven primary osseous hemangiopericytoma CT and MR scans were evaluated retrospectively. Both CT and MRI were available in four patients each. In three patients both imaging techniques were available. On CT primary hemangiopericytoma of bone presents as an expansive lytic lesion with bone destruction and inhomogeneous contrast enhancement. Magnetic resonance imaging depicts osseous hemangiopericytoma as hyperintense lesion on T2-weighted images with intermediate signal intensity on T1-weighted images. Curvilinear tubular structures of signal void in the tumor matrix on T1-weighted images and corresponding hyperintense structures on T2-weighted and on fat-suppressed short tau inversion recovery images were present in three patients. Although cross sectional imaging findings are non-specific, they add to the diagnosis and provide valuable information about the extent of bone destruction and local tumor spread in patients with primary osseous hemangiopericytoma. While CT demonstrates the extent of bone destruction best, MRI better visualizes medullary and soft tissue extension of the tumor. Curvilinear signal abnormalities support the diagnosis of hemangiopericytoma of bone. This imaging pattern is best visualized on fat-suppressed or contrast enhanced T1-weighted MR images. PMID- 11868081 TI - Hoffa's recess: incidence, morphology and differential diagnosis of the globular shaped cleft in the infrapatellar fat pad of the knee on MRI and cadaver dissections. AB - We frequently observed a fluid-like indentation at the inferior posterior margin of Hoffa's fat pad of the knee and sought to establish the incidence and differential diagnostic criteria of this cleft. In total, 133 MRI studies and 35 cadaver specimens were analyzed for the location, size, and shape of clefts at the inferior posterior margin of Hoffa's fat pad. The incidence of a fluid-like ovoid cleft on MR images was 13.5% and in cadavers 14.3%. The cleft was located just below the insertion of the infrapatellar synovial fold (plica synovialis infrapatellaris, ligamentum mucosum). More linear-shaped indentations at the posterior margin were visible in all patients and cadavers due to the horizontal course of the alar folds. A fluid-filled indentation within the inferior posterior margin of Hoffa's fat pad has to be expected in more than 10% of knee studies and should not be confused with tumors like ganglion cysts. We term this cleft the infrahoffatic recess. One hypothesis of its origin concerns the embryological regression process of the infrapatellar membrane into the infrapatellar synovial fold. It should not be confused with linear clefts due to the alar folds. PMID- 11868082 TI - MRI of the arthritic finger joints: synovial membrane volume determination, a manual vs a stereologic method. AB - The aim of this study was to compare the stereology method for estimation of synovial volume on MR images with manually outlining method in inflammatory joint diseases. As manual outlining of the synovial volume is too time-consuming a method for clinical use, a less time-consuming methods are needed. Post-contrast 3D T1-weighted turbo gradient MR images of the finger (metacarpophalangeal and interphalangeal) joints (14 joints) were acquired from 11 patients with rheumatoid arthritis ( n=8) and reactive arthritis ( n=3). Manually outlined volume was taken as a gold standard and compared with stereologic volume estimation on transverse 1-mm-thick slices. The mathematical basis of the stereologic volume estimator is based on a two-step procedure: a 2D nucleator is used for estimation of the area of the synovial membrane on an MR slice and unbiased estimates of volumes are obtained by Cavalieri's principle. The 2D nucleator estimates the area of any object irrespective of its size, shape, and orientation by measuring the distance between a "central point" in the object and the intersections between the object boundary and radiating test lines. The total volume of 154 finger joints was estimated. A significant correlation between manual and stereologic estimations of synovial volumes was found (Spearman rho=0.71, P=0.002). Ninety-five percent limits of agreement were +/- 5-6 ml (14 finger joints per patient). The time used for volume determination in a patient varied from 1 to 2.5 h by manual outlining and from 0.5 to 1 h by stereologic determination. Stereologic volume estimation can provide measures of synovial volumes comparable to the manual outlining method and is less time-consuming. Stereologic volume estimation seems to be a clinically useful method, especially if it is integrated in the MR unit's workstation. PMID- 11868085 TI - MRI of penile fracture: diagnosis and therapeutic follow-up. AB - A rupture of corpus cavernosum (CC) is a rare injury of the erect penis. The present study describes the role of MRI for diagnosis and follow-up of this injury. Four patients with clinically suspected acute penile fractures underwent MRI. Imaging findings were confirmed at surgery. In three patients, follow-up MRI was also available at 1, 6 and 16 weeks after surgical repair. In all patients pre-contrast T1-weighted images (T1WI) clearly disclosed ruptures of CC, which depicted as discontinuity of low signal intensity of the tunica albuginea (TA). Concomitant subcutaneous haematoma were well visualised both on T1-weighted (T1WI) and T2-weighted images, whereas haematoma in CC were optimally demonstrated on contrast-enhanced T1WI. On follow-up MRI all fractures presented similar healing process. Shortly after the repair, the tunical suture showed an increase in signal intensity on pre-contrast T1WI and was strongly enhanced with the administration of contrast material. Then the tear site gradually recovered low signal intensity on all spin-echo sequences by 4 months after surgery. These serial findings may suggest the formation of vascularised granulation tissue during cicatrisation. Magnetic resonance imaging is of great value for the diagnosis and follow-up in patients with penile fracture. PMID- 11868083 TI - Surface aneurysmal bone cysts: a pictorial review. AB - The aim of this article is to present the imaging features of surface aneurysmal bone cysts (ABCs). Twenty-three surface ABCs were identified from of a series of 144 histologically proven ABCs treated in our unit over the past 20 years. The surface ABCs showed a high female preponderance with a predilection for the forearm bones, tibia and femur. With the use of radiographs in all cases and CT and MR imaging in 18 cases it was possible to subdivide the series into subperiosteal (11 cases), cortical (8 cases) and mixed (4 cases) categories. The imaging features distinguishing the various categories are illustrated and the differential diagnosis is discussed. PMID- 11868084 TI - Unusual manifestation of vertebral osteoid osteoma: case report. AB - We report the case of a 64 year-old man with a clinical history suggesting a low thoracic-cord involvement, in which an unexpected vertebral osteoid osteoma was discovered. The patient underwent MRI of the thoraco-lumbar spine, which included sagittal and axial T1-weighted images, and sagittal double-echo T2-weighted images. Subsequently, CT scan was carried out with 2-mm-thick axial sections, aimed at T10 vertebra. Magnetic resonance imaging disclosed an extra-axial mass at T10 level. Computed tomography scan suggested an osteoid osteoma of the tenth thoracic vertebra, involving the lamina with marked sclerosis and prevalently endocanalar extension. Histology following surgical resection confirmed the diagnosis. In the reported case CT scan provided the correct pre-operative diagnosis of osteoid osteoma despite its unusual clinical--anamnestic presentation. Magnetic resonance imaging was useful in establishing the relationship of the neoplasm with the spinal cord. PMID- 11868086 TI - Ectopic pregnancy within a rudimentary horn in a case of unicornuate uterus. AB - We report the MRI features of two cases of unicornuate uterus and occluded rudimentary horn. In one patient pregnancy had occurred in the occluded horn, prompting to urgent resection. The second patient illustrates more conventional findings in occluded rudimentary horn. In both cases MRI was able to correctly characterise the nature of the developmental anomaly. Furthermore, on the basis of signal intensities, differentiation between distended lumen due to blood accumulation and amniotic sac in the case of pregnancy could be made. Because of its high accuracy in determining the type of anomaly present, the lack of ionizing radiation, and the ability to evaluate, with the exception of the fallopian tubes, the entire genitourinary tract, MR imaging is essential and plays a key role in the evaluation of women who are consulting for infertility. PMID- 11868087 TI - CT diagnosis of ureteral fibroepithelial polyps. AB - We report a case of fibroepithelial polyp of the ureter with serial CT examinations. Progressive growth of the fibroepithelial polyp was documented by CT within a period of 62 months. Excretory phase contrast-enhanced CT images accurately contributed to the diagnosis of ureteral fibroepithelial polyp and allowed limited surgical resection. Accurate imaging assessment of ureteral fibroepithelial polyps is essential for a conservative surgical approach and/or observation alone. PMID- 11868088 TI - Visualization of swallowing using real-time TrueFISP MR fluoroscopy. AB - The aim of this study was to evaluate the ability of different real-time true fast imaging with steady precession (TrueFISP) sequences regarding their ability to depict the swallowing process and delineate oropharyngeal pathologies in patients with dysphagia. Real-time TrueFISP visualization of swallowing was performed in 8 volunteers and 6 patients with dysphagia using a 1.5 T scanner (Magnetom Sonata, Siemens, Erlangen Germany) equipped with high-performance gradients (amplitude 40 mT/m). Image quality of four different real-time TrueFISP sequences (TR 2.2-3.0 ms, TE 1.1-1.5 ms, matrix 63 x 128-135 x 256, field of view 250 mm(2), acquisition time per image 139-405 ms) was evaluated. Water, yoghurt, and semolina pudding were assessed as oral contrast agents. Functional exploration of the oropharyngeal apparatus was best possible using the fastest real-time TrueFISP sequence (TR 2.2 ms, TE 1.1 ms, matrix 63 x 128). Increased acquisition time resulted in blurring of anatomical structures. As the image contrast of TrueFISP sequences depends on T2/T1 properties, all tested foodstuff were well suited as oral contrast agents, but image quality was best using semolina pudding. Real-time visualization of swallowing is possible using real time TrueFISP sequences in conjunction with oral contrast agents. For the functional exploration of swallowing high temporal resolution is more crucial than spatial resolution. PMID- 11868089 TI - Diagnosis of Heerfordt's syndrome by state-of-the-art ultrasound in combination with parotid biopsy: a case report. AB - The parotid gland is rarely affected by sarcoidosis (6% of all cases). If there is initial parotitis, and other major symptoms include fever, uveitis, or facial paralysis, the condition is referred to as Heerfordt's syndrome. The case presented here describes the diagnostic work-up and differentiation of swelling of the parotid gland by employing state-of-the-art ultrasound techniques in a 33 year-old patient with Heerfordt's syndrome. Color-coded duplex ultrasound demonstrated hypervascularization. Tissue harmonic and photopic imaging ultrasound additionally depicted hypoechoic, septated structural lesions of the glandular parenchyma with an optimized contrast. Since such structural changes are not a specific sonographic criterion for sarcoidosis, ultrasound-guided biopsy was performed for further clarification of the etiology of parotitis and for excluding lymphoma. In conclusion, state-of-the-art ultrasound with the option of obtaining a biopsy at low risk is a useful procedure for diagnosing unclear pathology of the parotid gland with rapid histological confirmation. PMID- 11868090 TI - Tuberous sclerosis: diffusion MRI findings in the brain. AB - Diffusion MRI has mainly been used for detection of acute ischemia, and for distinction of cytotoxic and vasogenic edema. We applied diffusion MRI in patients with tuberous sclerosis in order to evaluate diffusion imaging characteristics of parenchymal changes. Five children with known tuberous sclerosis were included in this study. The MRI examinations were performed on a 1.5-T MR unit. Diffusion MRI was obtained using the echo-planar imaging sequence. Apparent diffusion coefficient (ADC) values from the abnormal brain parenchyma were calculated directly from automatically generated ADC maps. Seven normal children were available for comparison. In this control group the mean ADC value of the normal white matter was 0.84 +/- 0.12 x 10(-3) mm(2)/s. In tuberous sclerosis patients the mean ADC value of the white matter hamartomas ( n=20) was apparently high (1.52 +/- 0.24 x 10(-3) mm(2)/s) compared with that of normal white matter. The ADC value of calcified hamartomas was "zero". The ADC value within a giant cell tumor was 0.89 x 10(-3) mm(2)/s, similar to that of normal cerebral white matter. The ADC maps were superior to b=1000 s/mm(2) (true diffusion) images with respect to lesion evaluation, and they provided mathematical information on tissue integrity. With respect to detection of the exact numbers and sizes of the parenchymal hamartomas fluid-attenuated inversion recovery images were superior to ADC maps. It is believed that diffusion MRI can be useful in evaluation of various parenchymal changes associated with tuberous sclerosis. Further studies on tuberous sclerosis, and on various brain lesions, would provide increasing data on this relatively new MRI sequence. PMID- 11868091 TI - Sneddon's syndrome: MR imaging findings. AB - Sneddon's syndrome is a systemic disease characterized by generalized livedo racemosa and cerebrovascular disease. We present a case of Sneddon's syndrome which has typical dermatological lesions and occipital infarct demonstrated by both CT and MRI. PMID- 11868092 TI - Cerebral and spinal cord involvement resulting from invasive aspergillosis. AB - Although central nervous system involvement in disseminated aspergillosis is known to occur in immunocompromised patients, particularly after bone marrow transplantation, localized involvement of the spinal cord is exceedingly rare. In this report we present and illustrate detailed imaging findings of central nervous system invasion by Aspergillus fumigatus in a 30-year-old woman, with emphasis on the spinal cord involvement. PMID- 11868093 TI - Tissue harmonic and contrast-specific imaging: back to gray scale in ultrasound. AB - The development of new US techniques that produce images based on nonlinear acoustic effects of US interaction with matter or microbubble contrast agents has opened new prospects for gray-scale US in native tissue and contrast imaging. Tissue harmonic imaging uses higher frequencies generated on propagation of the US beam through matter to improve image quality and resolve small anatomic structures and details, and is becoming a routine approach in US examination of many abdominal districts. Contrast-specific imaging techniques display enhancement of US agents in gray-scale with optimal contrast and spatial resolution, and offer high sensitivity either to microbubble movement or to microbubble destruction in dependence of the level of the applied acoustic peak pressure. Owing to the ability to exploit the microcirculation, contrast-specific techniques have enabled the evolution of contrast US from vascular imaging to the imaging of perfused tissues. Several studies have shown that these methods can substantially improve US detection and characterization of focal liver lesions, and promising results have been reported in other areas of investigation. This article reviews physical principles, technical issues, and clinical applications of tissue harmonic and contrast-specific imaging. It is foreseen that the new gray-scale US techniques will rapidly become a tool in numerous clinical scenarios. PMID- 11868095 TI - Anatomy of the minor fissure: assessment with high-resolution CT and classification. AB - The aims of this study were to investigate the anatomy of the minor fissure and its variations on high-resolution CT (HRCT) sections and to propose a detailed classification. The prospective study included 67 patients who were referred to CT for various indications. High-resolution CT examinations (1.5-mm collimation) were obtained through the region of the minor fissure. The CT scans were assessed for the presence, completeness, and configuration of the minor fissure. Various configurations of the minor fissure were classified into four major types, based on whether the highest portion of the middle lobe upper surface was medial (type I), lateral (type II), posterior (type III), or central (type IV). Minor fissure was identified in 65 (97%) of 67 patients, and absent in 2 (3%) cases. The fissure was incomplete in 35 (54%) of 65 patients. Type-I minor fissure is seen in 28 (43%) patients, type II in 22 (34%), type III in 5 (8%), and type IV in 2 (3%) patients. Because the majority of the fissure was absent in 8 (12%) of 35 patients with incomplete fissure, they were considered indeterminate. Comprehensive knowledge of the various configurations of the minor fissure is helpful in correct localization of a lesion and its extension. In equivocal cases, limited thin-section CT scans through the fissure delineate the anatomy more clearly and provide greater degree of precision in localizing pulmonary lesions. PMID- 11868094 TI - CT findings of leukemic pulmonary infiltration with pathologic correlation. AB - The aim of this study was to demonstrate the characteristic CT findings of leukemic pulmonary infiltration based on the pathologic findings. The CT findings of 11 leukemic patients with leukemic pulmonary infiltration were compared with those of 22 leukemic patients with other diseases as a control group. Evaluated pulmonary parenchymal CT findings included thickening of bronchovascular bundles and interlobular septa, prominence of peripheral pulmonary arteries, ground-glass opacities, air-space consolidation, and nodules. The CT-pathologic correlations for leukemic infiltration were evaluated in 7 patients. Frequent parenchymal CT findings were thickening of bronchovascular bundles (81.8%), prominence of peripheral pulmonary arteries (81.8%), and non-lobular and non-segmental ground glass opacities (90.9%). The first two findings were significantly more frequently observed in leukemic infiltration than in the control group, had good interobserver agreement, and corresponded pathologically to leukemic cell infiltration around the pulmonary arteries, bronchi, or bronchioles. Non-lobular and non-segmental ground-glass opacity corresponded to leukemic cell infiltration within alveolar spaces and septa adjacent to the pulmonary arteries or bronchi and also corresponded to hemorrhage, edema, or diffuse alveolar damage. Thickening of bronchovascular bundles and prominence of peripheral pulmonary arteries are CT findings suggestive for leukemic infiltration and correspond to peribronchovascular tumor extension. PMID- 11868096 TI - CT and MRI findings of mucin-containing tumors and pseudotumors of the thorax: pictorial review. AB - Bronchial mucus has tomodensitometric features and MR signal intensity similar to that of water. However, chronic entrapped mucus collections, due to water reabsorption and higher protein content, can have CT attenuation values higher than 20 and reaching even 130 HU. Higher protein concentration also causes a sensible reduction in T1 relaxation time. The demonstration of mucus within a mediastinal, bronchial or pulmonary lesion is an important diagnostic clue permitting remarkable shortening of the list of differential diagnoses. This article illustrates the CT and MR findings allowing correct characterization of the mucus-containing lesions of mediastinum, bronchi, and lung. PMID- 11868097 TI - Endotracheal neurofibroma in neurofibromatosis type 1: an unusual manifestation. AB - Tracheal involvement is an extremely rare manifestation in patients with neurofibromatosis type 1 (NF-1). We present a 33-year-old women with NF-1 suffering from progressive dyspnea. Multislice spiral CT revealed a neurofibroma located within the trachea with intratracheal extension. To our knowledge, this is the first report of an intratracheal neurofibroma which has been documented by CT. This indicates that multislice spiral CT allows accurate demonstration of localization and extent of this rare manifestation of neurofibromas. PMID- 11868098 TI - MR evaluation of coronary stents with navigator echo and breath-hold cine gradient-echo techniques. AB - The aim of this study was to evaluate coronary artery stents with MR. Thirty eight patients underwent MR imaging 48.1 +/- 6.6 days (range 38-60 days) after placement of 47 coronary stents of 11 different types, using navigator echo (NE) and cine gradient-echo (GE) techniques. For both sequences the low signal artifact was used to localize the stent, whereas the flow-related high signal before and distal to the stent was considered as a patency sign. Exercise electrocardiographic test (EET) had been performed 1-7 days before MR. No adverse event with possible relation to the MR examination was observed. All the stents were recognized as signal void with GE, and all but one with NE. Of the 2 patients with positive EET, the first one, with a stent on the left anterior descending coronary artery, presented low signal distal to the stent at both MR sequences, suggesting dysfunction [60% stenosis at conventional coronary angiography (CCA)]; the second one, with two sequential stents on the right coronary artery, presented lack of signal distal to the stents at both MR sequences, suggesting occlusion (97% stenosis at CCA). For the 44 remaining stents in 36 patients with negative EET, MR high signal before and distal to the stent suggested patency at both sequences. MR seems to be a safe and promising technique for non-invasive evaluation of coronary stents. PMID- 11868100 TI - Percutaneous thrombectomy: a review. AB - Percutaneous thrombectomy (PT) is an established technique for the removal of acute thrombus in occluded arteries, veins and vascular grafts. Percutaneous thrombectomy can be used as an adjunctive treatment to other methods of thrombus removal such as thrombolysis or as sole therapy. The two main methods are percutaneous aspiration thrombectomy in which thrombus is removed by suction with the aid of wide-bore catheters, and mechanical thrombectomy using a variety of automated devices to fragment or remove thrombus. Aspiration thrombectomy is often used as an adjunct to thrombolysis in acute arterial occlusion, or as salvage therapy to remove distal emboli following iliac or femoropopliteal angioplasty. Mechanical thrombectomy is useful for the treatment of thrombosed dialysis grafts and is being increasingly used for the treatment of massive pulmonary emboli and ileofemoral or ileocaval deep venous thromboses. PMID- 11868099 TI - Coronary artery aneurysm and type-A aortic dissection demonstrated by retrospectively ECG-gated multislice spiral CT. AB - The case of a 40-year-old male patient with a coronary aneurysm of the proximal left descending artery (LAD) combined with circumferential type-A dissection of the ascending aorta is reported. Computed tomography angiography of the coronary arteries was performed using multislice spiral computed tomography (MSCT) with retrospective ECG gating. Anatomical relations of the LAD aneurysm as well as the origin of the left coronary artery from the false lumen of the dissection were well depicted for planning of the surgical intervention using this new noninvasive imaging modality. PMID- 11868101 TI - Contrast-enhanced MR angiography of abdominal vessels: is there still a role for angiography? AB - The purpose of this review article is to describe recent advantages in contrast enhanced (CE) three-dimensional (3D) magnetic resonance angiography (MRA) in comparison with other vascular imaging techniques, and to discuss their current clinical applications for the imaging of abdominal vessels. Principles and technical considerations are presented and clinical applications are reviewed for different vascular diseases. In ruptured aortic aneurysms and acute dissections CT is the method of first choice. Contrast-enhanced 3D MRA can be well used for therapeutic planning and follow-up in patients with stable disease. A comprehensive MR examination including CE 3D MRA, MR urography and MR nephrogram has the potential to replace the conventional studies for the evaluation of renal vascular disease. It is an accurate method for imaging the origins of coeliac and superior mesenteric arteries, although the image resolution is too low for reliable assessment of the inferior mesenteric artery. Contrast-enhanced 3D MRA has emerged as the method of choice for studying the portal venous system in liver transplant recipients, in patients with portal hypertension and in cases with abdominal tumours for preoperative evaluation. Additional non-invasive flow measurements are useful in monitoring portal hypertension. The abdominal veins can be well imaged using unenhanced MR techniques. Imaging may be facilitated with intravascular contrast media. Contrast-enhanced 3D MRA can replace intra arterial DSA for diagnosis, therapy planning and follow-up in patients with abdominal vascular disease. Catheter-based arteriography will still be used for interventional procedures such as percutaneous transluminal angioplasty, stent placement and embolisation. PMID- 11868102 TI - Detachable balloon embolization of an aneurysmal gastroduodenal arterioportal fistula. AB - Extrahepatic arteriovenous fistulas involving the gastroduodenal artery and the portal venous system are rare and almost always a late complication of gastric surgery. Secondary portal hypertension and mesenteric ischemia may provoke abdominal pain, upper and lower gastrointestinal hemorrhage, diarrhea, and weight loss. Until recently, surgical excision has been the therapy of choice with excellent results. The authors report a case of gastroduodenal arterioportal fistula with a rare large interpositioned aneurysm in a cardiopulmonary compromised patient who was considered a non-surgical candidate. The gastroduodenal arterioportal fistula was occluded endovascularly by means of a detachable balloon. A survey of the literature of this rare type of arterioportal fistula is included. PMID- 11868103 TI - Accuracy of non-enhanced MRI to monitor histological lesion size during laser induced interstitial thermotherapy. AB - The purpose of this study was to assess the accuracy of non-enhanced MRI using a T1-weighted 2D turbo fast low-angle shot (FLASH) sequence during laser-induced interstitial thermotherapy (LITT) to determine histological lesion size of laser induced hepatic lesions. The LITT was performed on pig liver samples at various power settings and durations. For MR monitoring during and after LITT a T1 weighted 2D turbo-FLASH sequence was applied. Lesions seen by MRI during and after LITT were correlated with histological lesion size. Histologically, a core zone of complete tissue ablation close to the tip of the applicator could be differentiated from an adjacent transitional zone showing incomplete necrosis. Magnetic resonance imaging right at the end of LITT (i.e., with maximum heating effects) grossly overestimated the core zone but accurately described the transitional zone. Magnetic resonance imaging after cooling of the tissue (therefore showing structural as opposed to thermal changes) exactly depicted the core zone of complete tissue ablation. Non-enhanced MRI using a T1-weighted 2D turbo FLASH sequence strongly overestimates the histological lesion size during LITT; however, structural changes of the tissue seen after cooling accurately define lesion size in LITT. For clinical purposes the lesion geometry seen during MR monitoring should therefore well extend the tumor margins. PMID- 11868104 TI - The apple core sign. PMID- 11868105 TI - Self-citation: comparison between Radiologia, European Radiology and Radiology for 1997-1998. AB - Self-citation, considered as the number of times a paper cites other papers in the same journal, is an important criteria of journal quality. Our objective is to evaluate the self-citation in the official journal of the Spanish Society of Radiology (Radiologia), and to compare it with the European Radiology and Radiology journals. Papers published in Radiologia, European Radiology, and Radiology during 1997 and 1998 were analyzed. The Self Citation Index, considered as the ratio between self-references and total number of references per article, for the journals Radiologia (SCIR), European Radiology (SCIER), and Radiology (SCIRY), were obtained and expressed as percentages. Also, the number of references to Radiologia in European Radiology and Radiology papers were calculated. Stratification of the index per thematic area and article type was also performed. Mean SCIR, SCIER, and SCIRY values were compared with the ANOVA and the Student-Newman-Keuls tests. The self-citation index was statistically higher in Radiology (23.2%; p<0.0001) than in Radiologia (1.8%) and European Radiology (0.8%). There were no statistically significant differences between SCIR and SCIER indexes ( p=0.25). In the stratification per thematic areas and article type, self-citation in Radiology was statistically higher ( p<0.0001), with the only exception of "Radioprotection" area ( p=0.2), to SCIR and SCIER. Although there were no statistically significant differences, by thematic areas SCIR was always larger than SCIER, with the only exception of the "Genitourinary imaging" area, and by article type SCIR also went greater to SCIER, except in review articles. Radiologia, The Spanish official radiological journal, although not included in Index Medicus and its database Medline, had a larger number of self-citing than European Radiology in the period 1997-1998. PMID- 11868106 TI - Quiz case of the month. Elastrofibroma. PMID- 11868107 TI - A new CT sign of mediastinal lipomatosis? PMID- 11868108 TI - Comments on Dalla Palma et al.: Present-day imaging of patients with renal colic. PMID- 11868110 TI - Comments on F. Stacul: Current iodinated contrast media. PMID- 11868118 TI - Image Interpretation Session and Junior Image Interpretation Session, ECR 2002, Vienna. PMID- 11868117 TI - January 2002. PMID- 11868119 TI - Collaborative Care. PMID- 11868120 TI - Mandibular Advancement Modulates Respiratory-Related Genioglossus Electromyographic Activity. AB - The purpose of this study was to determine if mandibular advancement had any effects on the respiratory-related electromyographic (EMG) activity of the genioglossus (GG) muscle in normal adults for both the upright and supine positions. Spontaneous GG EMG activity during quiet nasal breathing was recorded in 5 mandibular positions: centric occlusion (CO), maximal protrusion (MAX), 25% (MAX25), 50%(MAX50), and 75%(MAX75) of MAX. The maximal GG EMG activities during inspiration (GGinsp) and minimal GG EMG activities during expiration (GGexp) were compared. The GGinsp and GGexp significantly increased with mandibular advancement in both body positions. Furthermore, there were significant differences in both GGinsp and GGexp between CO and more protruded mandibular positions. However, no significant differences were found in the GGinsp at MAX compared to that at MAX75, and in the GGexp at MAX compared to those at MAX75 and MAX50. These results suggest that augmentation of the respiratory-related GG EMG activity with mandibular advancement diminishes the propensity of the upper airway to collapse. Moreover, the lack of any remarkable difference between the GGinsp at MAX75 and that at MAX may be relevant to the effectiveness of oral appliances in the treatment of obstructive sleep apnea. PMID- 11868121 TI - Supine Cephalometric Analyses of an Adjustable Oral Appliance Used in the Treatment of Obstructive Sleep Apnea. AB - OBJECTIVE: To investigate the effects of the Klearway(TM) appliance on the upper airway in patients with obstructive sleep apnea (OSA) in the supine position. METHODS: Sixteen subjects (12 males and 4 females) were recruited on the basis of baseline polysomnography with a documented Apnea and Hypopnea Index (AHI) >15 per hour. A second overnight sleep study was performed for each subject with the appliance in place. Baseline supine cephalometry was performed for each subject before the initial insertion of the appliance, and follow-up supine cephalometry was undertaken with the appliance in place. RESULTS: The polysomnographic variables improved significantly, and the mean changes in overbite and overjet were 5.15 mm and 6.26 mm after insertion of the Klearway(TM) appliance. The supine sagittal cross-sectional areas of the pharynx and the tongue significantly increased, while the linear distance from the hyoid position to the mandibular plane or the RGN-C3 line significantly decreased after insertion of the appliance. The ratio of the vertical pharyngeal length to the sagittal cross sectional area of nasopharynx or tongue decreased significantly. When the subjects were evaluated on the basis of the after-insertion AHI, the group with good response (n = 11) was found to be significantly younger than the group with the poor response (n = 5). Similarly, the good responders revealed less prominent chins, larger tongue heights, and an increase in hypopharyngeal sagittal cross sectional area after insertion of the appliance. There was a significant correlation between the improvement in AHI (%) and the supine middle airway space (r = -0.52, p < 0.05). CONCLUSION: The mechanical effect of the Klearway(TM) appliance on the upper airway and the stabilization of jaw posture may be important determinants of the efficacy of the appliance. PMID- 11868122 TI - Neuropsychological and Behavioral Correlates of Obstructive Sleep Apnea Syndrome in Children: A Preliminary Study. AB - STUDY OBJECTIVES: The purpose of this pilot study was to evaluate a group of children with mild to moderate Obstructive Sleep Apnea Syndrome (OSAS) for baseline neurocognitive deficits and behavioral dysfunction. A subset of the sample were also reassessed, using the same test battery, after treatment with adenotonsillectomy. DESIGN: Baseline and post-treatment neuropsychological and behavioral assessment. SETTING: Pediatric sleep disorders clinic at a children's teaching hospital. PATIENTS: 18 children (12 males, 6 females, mean age 7.3 years plus minus 2.0) meeting polysomnographic criteria for OSAS underwent baseline assessment; 8 children (6 males, 2 females, mean age 8.4 years plus minus 2.6) also completed the post-treatment assessment phase. MEASUREMENTS: An age appropriate neuropsychological battery including measures of global cognitive functioning, language, executive functioning and attention, memory, visual perception/visual motor skills and motor skills; two parent rating scales of behavior. RESULTS: Modest impairments, largely in executive functioning/attention and motor skills, were found at baseline. Parents endorsed a variety of behavioral problems, especially somatic complaints and problems with learning. There appeared to be relatively little association between impairment and disease severity, although there was a trend for the children with less severe disease, who were also older, to have relatively more behavioral problems. Post treatment, there were modest improvements in executive functioning/attention and motor skills, as well as in parent-reported internalizing and externalizing behaviors. CONCLUSIONS: The preliminary results with a small sample suggest mild deficits in executive functions and motor skills in children with mild to moderate OSAS, with modest improvements in the same neuropsychological domains post-treatment. A variety of parent-reported behavioral problems were found at baseline, again with modest improvement post-adenotonsillectomy. PMID- 11868123 TI - Oral Appliance Therapy for the Management of Sleep Disordered Breathing: An Overview. AB - A burgeoning new arena in healthcare has opened to dental professionals with the potential to relieve the suffering of millions of people worldwide. Qualified dentists are increasingly being called upon to interface with the medical profession in an effort to manage the unstable upper airway during sleep. What has come to be called "oral appliance therapy" (OAT) involves the coordinated efforts of sleep physicians and the newly recognized sleep disorders dentist. PMID- 11868124 TI - Improvement in Daytime Sleepiness by the Use of an Oral Appliance in a Patient with Upper Airway Resistance Syndrome. AB - This case report details the treatment outcome of an oral appliance (OA) used in a patient suffering from excessive daytime sleepiness (EDS) caused by an upper airway resistance syndrome (UARS). The patient demonstrated significant improvement in the multiple sleep latency test (MSLT) and the maintenance of wakefulness test (MWT) after treatment with an OA. The presented case suggests that an OA may be considered as a further treatment option for UARS. PMID- 11868125 TI - Legal Perspective: Licensing and Liability Issues Regarding the Use of Oral Appliances in the Treatment of Obstructive Sleep Apnea. PMID- 11868126 TI - Cardiovascular Disease and Sleep-Disordered Breathing: The Bridge to the Mainland. PMID- 11868128 TI - Influence of Low-Flow Oxygen Supply on Sleep Architecture in Patients with Severe Heart Failure (NYHA III-IV) and Cheyne-Stokes Respiration. AB - Patients suffering from severe heart failure may develop breathing pattern disorders during sleep, especially in the form of Cheyne-Stokes respiration. Results may be severe disturbances in sleep architecture and worsening of hemodynamics and of prognosis of these patients. Causes of the periodic breathing disorders are probably hypocapnia, hypersensitivity of respiratory control centers, hypoxemia, and prolonged blood circulation time. This study examined the influence of different concentrations of continously administered oxygen during the nighttime on breathing pattern disorders, oxygen saturation, and sleep architecture in 65 patients with severe heart failure (NYHA III-IV). Fifty-two of 65 patients showed an improvement of sleep architecture. Total sleeping time increased significantly (p < 0.01). Fragmentations of sleep by arousals decreased ( p < 0.01); time of random eye movement (REM) sleep and non-REM sleep III and IV increased significantly. PMID- 11868127 TI - Sleep-Breathing Disorders and Heart Failure. AB - Cheyne-Stokes respiration is known to be associated with severe left heart failure. Because of severe desaturation, sleep fragmentation, arousals, and an increase in sympathetic activity, Cheyne-Stokes respiration may lead to a further impairment of cardiac function and to a worsening of quality of life. Although the pathology of Cheyne-Stokes respiration is not fully understood, enhanced chemoreceptor sensitivity, prolonged circulation time, as well as decreased pulmonary gas stores and increased ventilatory drive may be contributing factors. Therapeutic options include the improvement of cardiac failure; medical treatment, such as using theophylline; continous positive airway pressure ventilation; and low-flow oxygen supply. Because of severe cardiac insufficiency, change of endothoracic pressure may worsen the hemodynamic situation in some patients. Therefore, this form of treatment has to be used carefully. Another possible treatment is a low-flow oxygen supply, which will prevent severe desaturations. This therapeutic approach might be a good alternative to noninvasive ventilation. However, it is controversial whether oxygen supply will improve quality of sleep of the patients, even in long-term treatment. PMID- 11868129 TI - Influence of Orthotopic Heart Transplantation on Breathing Pattern Disorders in Patients with Dilated Cardiomyopathy. AB - Cheyne-Stokes respiration is well-known in patients with congestive heart failure. One of the causes might be a prolonged circulation time or diminished pulmonary gas stores. Improvement of cardiac function by heart transplantation might abolish Cheyne-Stokes respiration. We examined 29 male patients (age 24 to 63 years) with polysomnographically verified Cheyne-Stokes respiration before and 3 to 9 weeks after orthotopic heart transplantation. All patients suffered from congestive heart disease. Left ventricular ejection fraction (LVEF) was between 8 and 19%. We analyzed sleep stages, respiratory events, oxygen saturation, and cardiac function (echocardiography and ergometry). Before heart transplantation all patients showed periodic breathing (apnea-hypopnea index [AHI] 13.2-51.6/h). Oxygen saturation dropped to a minimum of 63%. Sleep was severely disturbed. After heart transplantation LVEF was significantly higher in all patients (xmean = 64%). Twenty-three of 29 patients showed no more evidence of periodic breathing. However, even with normalized cardiac function, 6 patients still suffered from Cheyne-Stokes respiration. Cheyne-Stokes respiration is often associated with dilated cardiomyopathy. Our results demonstrate that normalization of cardiac function improves Cheyne-Stokes respiration. Even after normalization of cardiac function, some patients suffer from Cheyne-Stokes respiration further on. We suggest that breathing control centers may be permanently damaged in these patients. PMID- 11868130 TI - Is Cheyne-Stokes Respiration Detrimental in Patients with Heart Failure? PMID- 11868131 TI - Broadening Our Therapeutic Spectrum. PMID- 11868132 TI - Videoendoscopic Assessment of Uncommon Sites of Upper Airway Obstruction during Sleep. AB - Patients with sleep-disordered breathing (SDB) suffer from repetitive upper airway occlusion. Various techniques have been described to assess the site of upper airway collapse. In most cases the soft palate or the base of tongue, or both, are the major levels of obstruction; rarely, the larynx, including the epiglottis, are found to be involved. We present five cases in which sleep videoendoscopy finally revealed the true mechanism of the inspiratory airway behavior. Two patients were sent to our service because of inefficacy of nasal ventilation therapy. One had a floppy epiglottis that was being sucked into the glottis. The other had a normal larynx during videoendoscopy. Instead, common pharyngeal collapse could be detected. Increasing the pressure normalized sleep, breathing, and videoendoscopy. The third patient showed apneas due to the adduction of the lateral parts of his omega-shaped epiglottis. In another case, inspiratory fluttering of the vocal cords caused snoring and arousals. The last case came in for bilateral vocal cord palsy with heavy nocturnal desaturations suggesting airway closure. Yet, the pharynx and larynx remained open throughout the night. Sleep videoendoscopy is a convenient method to define the level and mechanism of occlusion in obstructive SDB, particularly if the larynx is involved. It is useful to rule out airway compromise in case of recurrent nocturnal hypoxemias. Thus the results of sleep videoendoscopy have a strong therapeutical impact. PMID- 11868134 TI - The "Narco-sleep thing"--or are our patients getting biased? PMID- 11868133 TI - Long-Term Results of Maxillomandibular Advancement Surgery. AB - Surgery is a major modality in the treatment of obstructive sleep apnea syndrome (OSAS), and maxillomandibular advancement (MMA) has been shown to be the most effective surgical option. However, despite the successful short-term (6-9 months) results reported by various investigators, little is known of the long term clinical outcomes. A review of our long-term clinical results demonstrated that MMA achieves long-term cure in most patients. Aging and minor weight gain did not appear to have a significant adverse effect on the long-term results; however, major weight gain did seem to have a significant negative impact on the long-term outcomes. Therefore, long-term follow-up with proper counseling on weight maintenance or reduction is essential in improving long-term results. PMID- 11868135 TI - The importance of nasal resistance in obstructive sleep apnea syndrome: a study with positional rhinomanometry. AB - The importance of nasal obstruction in the pathogenesis of obstructive sleep apnea syndrome (OSAS) has not yet been totally defined. Numerous studies have reported an association between nasal obstruction and OSAS, but the precise nature of this relationship remains to be clarified. This study was undertaken to evaluate the prevalence of nasal obstruction disorders in a group of OSAS patients. For this purpose, we analyzed the nasal resistance of 36 OSAS patients by performing a traditional basal anterior active rhinomanometry test (AAR) and a positional AAR, with the patient in a supine position. Seven patients had a pathologic nasal resistance in the seated position that increased further in the supine position; 9 patients had normal resistance in the seated position but a pathologic resistance in the supine position. In 20 patients, nasal resistance was normal in both positions. No statistically significant differences in the degree of apnea/hypopnea index (AHI) was found between the 20 patients with normal positional AAR and the 16 with pathologic positional AAR (p = 0.13). Moreover, no statistically significant differences in the degree of AHI was found between the 7 patients with pathologic basal and positional AAR and the 9 patients with normal basal AAR and pathologic positional AAR (p = 0.38). PMID- 11868136 TI - Improving compliance with nasal CPAP and vigilance in older adults with OAHS. AB - The present study examined the efficacy of a cognitive-behavioral intervention at improving compliance with CPAP and vigilance in older adults with obstructive sleep apnea/hypopnea syndrome (OSAHS). Participants included 12 subjects who were randomized into one of two groups controlling for age, education, disease severity, and vigilance. The experimental group received two 45-min sessions designed to educate subjects on the consequences of OSAHS and the efficacy of CPAP. The control group received the same extent of therapist contact but did not receive information on OSAHS or CPAP. All subjects were administered a test of vigilance both before and after the study. Compliance data were collected using CPAP devices with internal microprocessors at were read at 1, 4, and 12 weeks after treatment initiation. The results showed that the experimental condition did not enhance compliance after 1 week of treatment but did so by the 12-week follow-up. Subjects in the experimental condition had a run time of 3.2-h per night longer than did those in the control group. Those using CPAP more regularly at 12 weeks also showed greater improvement on vigilance at follow-up. Performance on vigilance testing before the introduction of CPAP was predictive of CPAP use at 12 weeks. In conclusion, a modest cognitive-behavioral intervention may substantially increase CPAP use and vigilance in older adults. PMID- 11868137 TI - Treatment of snoring and obstructive sleep apnea with a mandibular protruding device: an open-label study. AB - The study objectives were to describe the changes in signs and symptoms of obstructive sleep apnea (OSA) and snoring using a mandibular protruding device (MPD) and to define the group of responders. The design of the study was a nonrandomized, open-labeled, prospective clinical study with a duration of 6 months. The setting was a hospital sleep laboratory and an ear, nose, and throat and dental outpatient clinic. Patients included 35 individuals treated: 22 with a diagnosis of OSA and 13 who snored without apnea. The intervention before MPD prescription entailed a medical examination, a one-night somnographic registration, a questionnaire, and a clinical jaw function examination. All procedures were iterated at the 6-month follow-up. When the patients used the MPD, the subjective symptoms decreased significantly (p < 0.001). Twenty-four of the 35 patients were classified as responders, i.e., reduced their symptoms > or = 50%. In the groups of OSA patients, the mean peak intensity of the snoring sound decreased from 71.6 dB to 62.0 dB (p < 0.001) and in the group of snorers, from 63.5 dB to 57.5 dB (p < 0.05). Eighteen of the 22 OSA patients decreased their ODI values by > or = 50% (responders). The mean ODI decreased from 15.4 to 3.5 (p < 0.001) and the blood oxygen saturation, SaO(2) nadir, increased from mean 81.9 to 85.7 (n.s.). Most patients with tongue base hyperplasia were classified as responders to the MPD treatment. In conclusion, a 6-month perspective shows that the MPD could be an effective treatment in reducing signs and symptoms of OSA and/or snoring, the compliance was acceptable and only minor adverse events occurred. PMID- 11868139 TI - Oral appliance therapy: the value of dental expertise. PMID- 11868138 TI - Morbidity of obstructive sleep apnea in children: facts and theory. AB - Sleep-disordered breathing (SDB) is a frequent, albeit underdiagnosed, problem in children. If left untreated, SDB may lead to substantial morbidities affecting multiple target organs and systems. This review provides a detailed and current description of the current status of our understanding of SDB-associated morbidity in children, and provides recommendations of future research directions necessary for increasing our knowledge and awareness on the short- and long-term consequences of SDB during childhood. PMID- 11868140 TI - Incorporating inheritance into models for understanding ventilatory behavior. AB - Ventilation and its components (frequency and tidal volume) appear to be determined to a significant extent by inheritance. Gene manipulation, gene identification, and functional genomics now offer powerful tools to identify the strength and mode of inheritance for ventilatory behavior under steady-state and non-steady-state conditions, in health and in disease. Conscious integration of genetic principles into existing explanatory models may increase the likelihood of detecting traits that correlate with protein systems responsible for the structures and the functional components of respiration. PMID- 11868141 TI - Concerning the sleep curriculum in a pulmonary training program. PMID- 11868142 TI - Left ventricular dysfunction, pulmonary hypertension, obesity, and sleep apnea. AB - The purpose of this study was to determine the frequency of central and obstructive sleep apnea in adult patients who have echocardiographic evidence of left ventricular dysfunction and pulmonary hypertension. Subjects with left ventricular dysfunction, pulmonary hypertension (pulmonary artery systolic pressure >30 mm Hg) and no lung disease were evaluated for risk factors associated with pulmonary hypertension. Of eight eligible adults, six completed the study. Subjects were from suburban and inner city family practices. Spirometric assessment, pulse oximetry on room air, rheumatologic evaluation, polysomnography, and additional history were taken. All six subjects had sleep apnea (apnea-plus-hypopnea index, or AHI, > or = 20): obstructive, central, or mixed. All were obese, and almost all the subjects had a restrictive pattern on spirometry, which is consistent with obesity. All had a pulmonary artery systolic blood pressure of 35 mm Hg or greater. None had daytime hypoxemia or collagen vascular disease, and none had ever used appetite suppressants. This study found a strong association between pulmonary hypertension and obstructive or central sleep apnea in obese patients with congestive heart failure (CHF). We propose that a pulmonary artery systolic pressure of 35 mm Hg or greater in ambulatory patients with CHF may signify an increased risk of sleep apnea. PMID- 11868143 TI - Snoring and sleep disturbance among children from an orthodontic setting. AB - Studies of snoring and sleep disturbance in the United States have been predominantly in clinic-based settings of children suspected to have obstructive sleep apnea hypopnea syndrome. Therefore, the purposes of this study were to utilize an orthodontic setting in which healthy children were seen regularly to study the prevalence of snoring and sleep disturbance among 405 children aged 6 to 17 years of age and to identify specific sleep behavior patterns associated with the increased odds of snoring. A sleep behavior questionnaire was administered to the child's parent or guardian. The questionnaire responses were analyzed using chi(2) analysis, and factor analysis was used to extract meaningful domains. The selected domains were used later in a logistic model to calculate the odds of snoring. The results indicated that 17% of the children habitually snored. The odds of snoring were approximately three times greater among mouth breathers and children who slept with their head tipped back and 1.5 times greater among those with morning headaches and frequent coughs and colds. In conclusion, snorers have significantly more sleep behavior problems than do non-snorers. PMID- 11868144 TI - Cervical positioning for reduction of sleep-disordered breathing in mild-to moderate OSAS. AB - The objective of this study was to assess whether cervical positioning could improve mild to moderate cases of the obstructive sleep apnea syndrome (OSAS). Eighteen subjects recruited from a tertiary sleep disorders clinic population with mild to moderate cases of OSAS were evaluated using a custom-fitted cervical pillow designed to increase upper airway caliber by promoting head extension. The subjects used their usual pillows during two consecutive recorded baseline nights in our laboratory. They then used the cervical pillow for 5 days at home and returned for 2 consecutive recorded nights at our laboratory to use the cervical pillow. During the nights in our laboratory, the subjects completed questionnaires, were videotaped to record head and body position, and had full polysomnography. The subjects had a significant trend toward improvement in their respiratory disturbance indices with use of the cervical pillow, despite spending more time in the supine position and having similar amounts of REM sleep in the baseline and experimental conditions. They also had nonsignificant trends toward improvements in their sleep efficiency and subjective depth of their sleep as well as significantly fewer arousals and awakenings in the experimental compared with the baseline condition. We propose that cervical positioning (i.e., head extension) with a custom-fitted cervical pillow provides a simple, noninvasive, and comfortable means of reducing sleep-disordered breathing in patients with mild to moderate OSAS. PMID- 11868145 TI - Subgrouping persons with snoring and/or apnea by using anthropometric and cephalometric measures. AB - Craniofacial and anthropometric characteristics are identified risk factors for obstructive sleep apnea syndrome (OSAS). Phase 1 of this study used cephalometric radiographs to record craniofacial measurements from 60 OSAS subjects with a respiratory disturbances index (RDI) of 20 or greater (group A) and 60 subjects with a history of loud snoring and an RDI less than 20 (group B). From this data set, a craniofacial risk index (CRI) was constructed using age, body mass index (BMI), and 14 cephalometric measures previously reported to be associated with OSAS. A separating boundary (CRIS) was established by using discriminant analysis to differentiate between the two groups. All measurements were determined by an investigator who was blinded to the subjects' RDI score. Phase 2 used a second sample of 19 group A and 47 group B subjects to test the ability of the CRI derived from the first sample to classify subjects in this second sample into the correct age group. The CRI was able to classify correctly 72.3% (34 of 47) of the group A subjects when all variables were used in the discriminant model. Using only four variables (age, BMI, hyoid mandibular plane distance, and tongue length) selected by the stepwise method, 72.3% (34 of 47) of the group B subjects and 78.7% of group A subjects were classified correctly. These results suggest that a stepwise CRI could be used to classify heterogeneous groups of individuals with increased RDI into subgroups with varying degrees of anatomic risk for disease. Such subgrouping by anatomic risk could be important in determining the pathophysiology of OSAS because it is likely that differences in upper airway anatomy among individuals interacts with a variety of other factors to produce clinical illness. PMID- 11868146 TI - The upper airway resistance syndrome masquerading as nocturnal asthma and successfully treated with an oral appliance. AB - Over the past 10 years, our ability to recognize, treat, and identify the morbidity associated with the upper airway resistance syndrome (UARS) has improved vastly. The diagnosis of this syndrome is dependent on a high degree of clinical suspicion, and in the presence of an already known pulmonary disease such as asthma, the identification of UARS may be elusive. Treatment of this condition has received more recent attention in the literature, with oral appliance therapy as a viable treatment option in place of the usual positive pressure ventilation devices. PMID- 11868148 TI - What is a necessary knowledge base for sleep professionals? AB - Sleep medicine is multidisciplinary, and sleep medicine professionals should be trained to evaluate and treat all 88 sleep disorders. Sleep medicine specialists require a fund of knowledge that goes beyond what is obtained during a pulmonary fellowship. Skills required for a pulmonary sleep professional include: sleep medicine, neurobiology, psychiatry, neuro-psychology, neurology, pediatrics, and even limited exposure in otolaryngology, oral maxillofacial surgery, and dentistry. There is a paucity of published information concerning curricular requirements. Required skills for a sleep professional include proficiency in the clinical skills of sleep medicine as well as the technical skills of polysomnography. There is a very large knowledge content area requirement in both the basic sciences of sleep and the clinical aspects of sleep medicine. There are also important clinical skills content areas. As with all medical professionals, sleep professionals should have the highest ethical standards and a strong sense of responsibility toward their patients. A sleep medicine professional also has to be knowledgeable about administrative and legal aspects specific to sleep medicine. This essay reviews a sleep professional knowledge base model with emphasis on the requirements for a pulmonary sleep professional. PMID- 11868147 TI - Application of telemedicine to sleep medicine. AB - Polysomnograms (PSGs) in specialized sleep centers with physicians and technicians trained in sleep medicine are still considered to be the most accurate form of diagnosis in patients with sleep disorders. But they are also very costly. Internet online transfer of PSGs from nonstaffed to very well staffed sleep centers might be a solution in the future to reduce costs. PMID- 11868149 TI - Continuous positive airway pressure and obstructive sleep apnea in an Hispanic population. AB - OBJECTIVES: We sought to explore the relationship between the diagnosis of hypertension and obstructive sleep apnea (OSA) in a Hispanic population, describe the effect of continuous positive airway pressure (CPAP) on blood pressure regulation, and assess the effect of CPAP on quality of life. DESIGN: A retrospective, recall interview study. PARTICIPANTS: Patients enrolled at the Home Oxygen Program of the San Juan V.A. Medical Center with the diagnosis of OSA and treatment with CPAP. MEASUREMENTS: The Calgary Sleep Apnea Quality of Life Index was administered to all patients after informed consent. Information regarding co-morbid conditions and fluctuations in blood pressure and anthropometric variables were obtained on a follow-up evaluation. RESULTS: After excluding for the use of antihypertensive medications, weight, and age, a 10% decrease in mean arterial pressure (MAP) from 100 mm Hg to 92 mm Hg was observed in an average of 40 months of therapy (p < 0.05). With the Calgary Quality of Life Index, 67% of the patients reported an extreme improvement in their quality of life. Compliance with CPAP therapy correlated with improved quality of life (r = 0.33, p < 0.015). CONCLUSIONS: In this pilot study, there appears to be a correlation between our intervention and decline of blood pressure, independent of body weight, age, or medication usage. CPAP treatment is an effective modality in improving symptoms and quality of life. PMID- 11868150 TI - Knowledge of sleep apnea in a sample grouping of primary care physicians. AB - The purpose of this pilot study was to examine four groups of primary care physicians' knowledge of sleep apnea. METHODS: Using a 36-item questionnaire, we investigated how cognizant primary care physicians in Ontario, Canada, were of sleep apnea and its different symptoms. The questions covered incidence, diagnosis, treatment, and medical and social ramifications of sleep apnea. Sleep apnea surveys were administered to small groups of primary care physicians attending educational conferences or were distributed by mail to physicians who had previously referred patients to the sleep clinic. RESULTS: A total of 151 physicians responded to the survey. An overall average score of 69% was obtained on the questionnaire. CONCLUSIONS: This score suggests that the physicians sampled in this pilot study are relatively under-informed about the clinical features and medical and social ramifications associated with sleep apnea. PMID- 11868151 TI - Medical student competence in eliciting a history for "chronic fatigue". AB - PURPOSE: We report an observational study of medical students' abilities in taking a complex history for which sleep disorders is one of several possible conditions. METHODS: Students are observed taking a focused history from a simulated patient whose chief complaint is "I am tired. I cannot get anything done." Nine groups of students (n = 360) completing the internal medicine core clerkship were evaluated by one of three examiners. Students received full, partial, or no credit for each item on a uniform behavioral checklist, which included prompts for common medical and psychiatric disorders associated with chronic fatigue. RESULTS: Observed means were lowest for items pertaining to sleep behaviors and head trauma. Fewer than half of the students inquired about whether or not the person had difficulty falling asleep at night, family history of sleep apnea, and frequency and length of naps. In contrast, the majority of students inquired about heart disease, metabolic disorders, the use of illicit drugs, alcohol consumption, and the taking of medications. Examiners accounted for a significant source of variance in scores; yet the station discriminated among top and bottom students as measured by the Objective Structured Clinical Examination (OSCE) overall. No statistically significant differences were observed on the basis of clerkship site, primary care versus traditional-track students, time of year, or gender. CONCLUSION: A majority of students do not adequately cover issues relevant to sleep in contrast to other associated disorders when taking a focused history for chronic fatigue. PMID- 11868152 TI - Misinterpretation of sleep-breathing disorder by periodic limb movement disorder. AB - We report a case of misinterpretation of sleep-disordered breathing due to periodic limb movement disorder. A 67-year-old man was diagnosed with sleep disordered breathing and subsequently placed on treatment with nasal continuous positive airway pressure (CPAP). The initial diagnostic evaluation did not include measurement of anterior tibialis electromyogram. The respiratory disturbance index of the initial evaluation was 23. After a brief period of nasal CPAP use, the patient discontinued the treatment because no significant change in daytime alertness was noted and signs of CPAP-related insomnia appeared. The patient was restudied polysomnographically with monitoring of anterior tibialis electromyograms. This study identified 392 leg movements of which 65% were associated with brief EEG arousal from sleep. Double-blind analysis of respiratory disturbance and leg movements yielded a total number of 360 arousals in the overnight recording. Eighty-five percent of all respiratory events could be associated with central hypoventilation following periodic limb movement associated EEG arousal. No significant hypoxia was recorded with these events. We hypothesize that chemoreceptor stimulation secondary to EEG arousal during sleep is responsible for this central hypoventilation. This case report highlights that recording and scoring of leg movements must be an integral part of polysomnographic evaluations. PMID- 11868153 TI - Is adjustability advantageous in mandibular advancement appliances in the treatment of sleep-disordered breathing? AB - Mandibular advancement appliances (MAD) that can adjust the amount of mandibular protrusion in response to treatment results are becoming popular with dentists who treat sleep-disorder breathing. Clinicians commonly assume that they are more efficacious and more comfortable to wear than monoblock appliances. An adjustable MAD was compared with one that stabilized the mandible at 70% of the protrusive path. There was not a significant difference in their overall efficacy. The adjustable MAD appeared to be more effective in the more severe cases, presumably due to the greater mandibular advancement; the nonadjustable MAD appeared to be more effective in the less severe cases, presumably due to the greater mandibular stabilization. Existing research was examined and biomechanical principles were considered. Insufficient research exists to definitely evaluate the pros and cons of adjustability vs. stability. There does not appear to be sufficient evidence to support the assumption that adjustable MADs are more efficacious or more comfortable to wear than monoblock appliances. PMID- 11868154 TI - Meet the National Center on Sleep Disorders Research and the National Heart, Lung, and Blood Institute: research opportunities in sleep medicine. PMID- 11868155 TI - Employee/employer interactions and responsibilities with special reference to genetically related sleep disorders. PMID- 11868156 TI - Higher prevalence of smoking in patients diagnosed as having obstructive sleep apnea. AB - Nightly nicotine withdrawal as well as other respiratory and pulmonary effects of smoking may result in sleep-disordered breathing, especially obstructive sleep apnea (OSA). We hypothesize that there is higher prevalence of smoking in patients with OSA. We also hypothesize that smoking is an independent risk factor for OSA. The aim of this study is to determine whether there is a higher prevalence of smoking in patients with OSA compared with patients who do not have OSA. To investigate this, we randomly selected a group of 108 patients who were diagnosed as having OSA, defined by an apnea-hypopnea index (AHI) of greater than 10 events per hour. We compared their smoking history with another randomly selected group of 106 patients without OSA, defined by an AHI of less than five events per hour. The prevalence of smoking in patients with OSA was found to be 35%, whereas it was only 18% in patients without OSA. Logistic regression analyses were performed to investigate the effects of smoking while adjusting for age, gender, body mass index (BMI), and number of alcoholic drinks per week. While holding fixed the BMI, gender, age, and number of alcoholic drinks per week, current smokers were found to be 2.5 times more likely to have OSA than former smokers and nonsmokers combined (odds ratio = 2.5, CI 1.3-4.7, p = 0.0049), and 2.8 times more likely to have OSA than former smokers alone (odds ratio = 2.8, CI = 1.4-5.4, p = 0.0028). Adjusted for BMI, gender, age, and number of alcoholic drinks per week, former smokers were not more likely than never smokers to have OSA (odds ratio = 1.2, CI = 0.55-2.7, p = 0.64). We conclude that cigarette smoke may be an independent risk factor for OSA in this referral population. PMID- 11868158 TI - Sleepiness, troika of consciousness cycle, and the Epworth sleepiness scale. AB - Excessive daytime sleepiness (EDS) is an important indicator when diagnosing sleep-disordered breathing and evaluating its treatment results. However, there appears to be some confusion as to what exactly is sleepiness; Dorland's Illustrated Medical Dictionary does not help. The medical literature was reviewed in order to assemble a schematic model that would suggest a definition of sleepiness and how it can be measured. The derived model is entitled the troika of consciousness cycle (TCC). It assumes that the presence of wakefulness, nonrapid eye movement sleep (NREMS), and rapid eye movement sleep (REMS) is determined by the interactions of four drives: two promoting wakefulness and one each for the two sleep states. The TCC illustrates that inadequate sleep results in sleep debt, but that sleepiness is determined solely by the nearness of the secondary wake drive line to the NREMS drive line. Contact of these lines indicates dozing, a change in consciousness state, an observable event. The probability of this event may be defined as objective sleepiness; this is what the Epworth sleepiness scale (ESS) attempts to measure. Studies indicate that the ESS can determine EDS with greater sensitivity and selectivity than either the multiple sleep latency test or the maintenance of wakefulness test. PMID- 11868157 TI - Mortised genioplasty in the treatment of obstructive sleep apnea: an historical perspective and modification of design. AB - We describe a modified technique for mortised genioglossus advancement for treating obstructive sleep apnea and review the history of osteotomies in this region. This new osteotomy technique allows for greater soft tissue advancement of the hypopharyngeal region. Anatomical data from a previous study were used to evaluate the dimensions of the anterior mandible and design an osteotomy that overcomes shortcomings of previous designs. These anatomic measurements enabled us to estimate the size and formulate a design utilized in the anterior mandible for the treatment of obstructive sleep apnea. We believe this design offers the greatest amount of muscular advancement by including genioglossus, geniohyoid, digastric, and mylohyoid. This advancement results in increasing the posterior airway space by volumetric expansion. Custom-designed fixation was utilized to increase stability laterally and decrease the risk of mandibular fracture. The design should be a significant aid in reconstruction of the hypopharyngeal airway in patients with obstructive sleep apnea. PMID- 11868159 TI - New technologies to detect static and dynamic upper airway obstruction during sleep. AB - Increase in upper airway resistance is the main patho-physiological feature in the obstructive breathing disorders during sleep. Upper airway events may be divided into two main groups: static obstruction (apneas) and dynamic obstruction (hypopneas, flow limitation, and snoring). This classification is useful to provide better information about the patho-physiological mechanisms of obstruction and to better define the diagnostic tools necessary for detecting abnormal respiratory events during sleep. Detection of dynamic obstruction requires sensors with a good frequency response. As thermistors have a poor dynamic response, they are not efficient in detecting the dynamic obstruction but are good enough to detect static obstruction. Nasal prongs (NP) connected a to pressure transducer and the impedance signal measured by the forced oscillation technique (FOT) are relatively new tools to noninvasively investigate dynamic upper airflow obstruction during sleep. FOT provides a direct index of the magnitude of airway obstruction and, therefore, of the upper airway patency, even under conditions of no flow (apneas). NP are aimed at assessing flow. Thus, both techniques have a different scope. The main advantages of NP are that they are easy to use and do not require sophisticated technology, while FOT needs a more complex instrumentation. For clinical routine studies NP are probably the best and simplest method for assessing the different respiratory events during sleep. However, FOT would be particularly useful in selected applications such as assessing upper airway patency in some central apneas; interpreting the irregular pattern of breathing during REM sleep; in better characterizing the inspiratory flow-limited breaths classified as intermediate; and in studying upper airway mechanics. PMID- 11868160 TI - A mutation in the SOS1 gene causes hereditary gingival fibromatosis type 1. AB - Hereditary gingival fibromatosis (HGF) is a rare, autosomal dominant form of gingival overgrowth. Affected individuals have a benign, slowly progressive, nonhemorrhagic, fibrous enlargement of the oral masticatory mucosa. Genetic loci for autosomal dominant forms of HGF have been localized to chromosome 2p21-p22 (HGF1) and chromosome 5q13-q22 (HGF2). To identify the gene responsible for HGF1, we extended genetic linkage studies to refine the chromosome 2p21-p22 candidate interval to approximately 2.3 Mb. Development of an integrated physical and genetic map of the interval identified 16 genes. Sequencing of these genes, in affected and unaffected HGF1 family members, identified a mutation in the Son of sevenless-1 (SOS1) gene in affected individuals. In this report, we describe the genomic structure of the SOS1 gene and present evidence that insertion of a cytosine between nucleotides 126,142 and 126,143 in codon 1083 of the SOS1 gene is responsible for HGF1. This insertion mutation, which segregates in a dominant manner over four generations, introduces a frameshift and creates a premature stop codon, abolishing four functionally important proline-rich SH3 binding domains normally present in the carboxyl-terminal region of the SOS1 protein. The resultant protein chimera contains the wild-type SOS1 protein for the N-terminal amino acids 1-1083 fused to a novel 22-amino acid carboxyl terminus. Similar SOS1 deletion constructs are functional in animal models, and a transgenic mouse construct with a comparable SOS1 chimera produces a phenotype with skin hypertrophy. Clarification of the functional role of this SOS1 mutant has implications for understanding other forms of gingival fibromatosis and corrective gingival-tissue management. PMID- 11868162 TI - Long-term care must not be the ugly sister. PMID- 11868163 TI - Policies are needed to clarify DNR debate. PMID- 11868164 TI - Siamese twins: both lives must be equally respected. PMID- 11868161 TI - Newfoundland rod-cone dystrophy, an early-onset retinal dystrophy, is caused by splice-junction mutations in RLBP1. AB - Some isolated populations exhibit an increased prevalence of rare recessive diseases. The island of Newfoundland is a characteristic geographic isolate, settled by a small number of families primarily during the late 1700s and early 1800s. During our studies of this population, we identified a group of families exhibiting a retinal dystrophy reminiscent of retinitis punctata albescens but with a substantially lower age at onset and more-rapid and distinctive progression, a disorder that we termed "Newfoundland rod-cone dystrophy" (NFRCD). The size of one of these families was sufficient to allow us to perform a genomewide screen to map the NFRCD locus. We detected significant linkage to markers on the long arm of chromosome 15, in a region encompassing RLBP1, the gene encoding the cellular retinaldehyde-binding protein. Previously, mutations in RLBP1 have been associated with other retinal dystrophies, leading us to hypothesize that RLBP1 mutations might also cause NFRCD. To test this hypothesis, we sequenced all coding exons and splice junctions of RLBP1. We detected two sequence alterations, each of which is likely to be pathogenic, since each segregates with the disease and is predicted to interfere with mRNA splicing. In contrast to some previously reported RLBP1 mutations, which yield a protein that may retain some residual activity, each NFRCD mutation is likely to give rise to a null allele. This difference may account for the severe phenotype in these families and exemplifies the molecular continuum that underlies clinically distinct but genetically related entities. PMID- 11868165 TI - Case 34: patient consent. Nurse who carried out manual evacuations without consent. PMID- 11868166 TI - Developing an oral assessment and intervention tool for older people: 1. AB - This is the first in a series of three articles relating to oral care for older people. The first article underlines the importance of oral assessment and evidence-based intervention for older people. It describes influences which can affect older people's ability to maintain oral healthcare practices. There is a need to examine closely issues surrounding poor oral hygiene practices and their associated ramifications for the older person. Identifying and implementing an appropriate oral assessment tool for older people should provide the foundation for the maintenance and improvement of standards of oral care practices within the clinical practice setting. The second article critically analyses oral care interventions; the third article critiques a number of published oral assessment tools with a view to developing and implementing a workable oral assessment tool for the practice setting. PMID- 11868167 TI - Cost benefits of two dressings in the management of surgical wounds. AB - In this prospective randomized controlled trial of the cost benefits of the choice of dressings in acute surgical wounds left to heal by secondary intention, patients had their wounds dressed with either a traditional dressing (ribbon gauze soaked in proflavine) or a modern hydrofibre dressing. Results showed that the hydrofibre dressing, although more expensive than the ribbon gauze, facilitated an earlier discharge from hospital (P = 0.001). The total cost of the patient episode was less in the hydrofibre group (P = 0.01). In an average UK health authority of 300,000 population, 100 bed days a year could be saved releasing an overall potential saving of 55,000 Pounds. A modern hydrofibre dressing allows more effective use of scarce hospital beds and precious financial resources, while still maintaining high quality patient care. It is important that clinical specialists and purchasers of health care should be aware of this clinical and cost-effective advance in the management of acute surgical wounds. PMID- 11868169 TI - Responsibility for controlled drugs in operating departments. AB - The difficulties experienced with the recruitment and retention of registered nurses has made it necessary for NHS trusts to review the traditional role of nurses and to transfer some of their duties to other groups of staff in order to maintain services. For example, in the operating department, operating department assistants and operating department practitioners (ODAs/ODPs) are undertaking some of the duties that were previously the responsibility of nurses. This article presents the findings of a survey to determine the extent to which ODAs/ODPs have access to controlled drugs in operating theatres. The survey was initially undertaken by the author in Welsh hospitals during 1998. The mixed responses generated discussion in the Duthie Review Committee (Committee Chairman, personal communication, Royal Pharmaceutical Society of Great Britain (RPSGB), 1999) and led to a collaborative exercise based on a random selection of NHS trust hospitals in England, Scotland and Northern Ireland to obtain a broader data across the UK. Despite a clear legal position concerning possession, the survey showed that practice varied widely and that some trusts may be contravening the Misuse of Drugs Regulations 1985 in order to keep the service running when registered nurses are not available. PMID- 11868168 TI - Nutrition and the life cycle. 1: Maternal nutrition and pregnancy. AB - Babies who are smaller than average at birth are more likely to face health problems during their adult life. A major influence on the size of the baby is maternal nutrition. Maternal energy requirements are not greatly increased, but other nutrients have been shown to influence fetal wellbeing; these include folate, riboflavin and some polyunsaturated fatty acids. Other nutrients that may enhance fetal welfare are ascorbic acid, zinc, and magnesium, and there are probably many more. Nurses, midwives, and health visitors need to be aware of the nutritional needs of pregnant women, and of the need to assist women to meet these needs. This is the first in a series of articles reviewing nutritional needs to maintain health during progressive stages of the life-cycle. This article discusses issues surrounding nutrition in pregnancy. Subsequent articles will review nutritional needs in young children, school-age children, adults, and older adults. PMID- 11868170 TI - Patients' attitudes towards the use of gloves by healthcare staff. AB - There is a concern among healthcare providers regarding the use of gloves in non mandated clinical situations and its impact on patient sensitivities. A convenience sample of 76 Israeli hospital inpatients were surveyed to measure patients' attitudes towards the use of gloves by healthcare providers. The Patient Sensitivity Questionnaire (PSQ) was developed and used to collect the data. Overall, patients were found to have a positive attitude towards the use of gloves in most patient care situations. Significant differences in patient attitudes towards glove use were found based on country of origin and number of hospital admissions in the previous year. The study concluded that hospitalized patients, in general, support healthcare providers' use of gloves. In addition, subjects agreed with the (American) Occupational Safety and Health Administration regulations regarding glove use. PMID- 11868171 TI - Preventing fungal infections in immunocompromised patients. AB - The incidence of systemic fungal infections in immunosuppressed patients has increased in the past two decades (Fridkin and Jarvis, 1996). Bone marrow transplant recipients and patients with prolonged neutropenia are most at risk, particularly when environmental factors increase the presence of fungi and fungal spores. Preventive measures include air filtration, handwashing, exclusion of flowers and some foodstuffs and use of chemoprophylaxis. In April 1999, a meeting was held under the auspices of the European Group for Blood and Marrow Transplantation (UK) Nurses and Allied Professionals Group to discuss the role of nurses in the management of patients at risk of systemic fungal infection. Issues discussed included: the need for nurses to recognize risk factors for systemic fungal infection; the role of nurses in giving feedback to patients; and the use of antifungal agents and associated problems, such as non-compliance and side effects. PMID- 11868172 TI - Pathways for continence care: development of the pathways. AB - This article is the second in a series of three covering a project into the use of care pathways for continence care undertaken by the authors. Loddon NHS Trust, Wiltshire and Swindon Healthcare NHS Trust and Salisbury Healthcare NHS Trust collaborated and supported their continence advisers in moving from financially driven assessment data to writing evidence-based care pathways and supporting patient information. The first article (Vol 9(9): 590-6) described the issues facing the continence advisers and the background to their decision to use full evidence-based care pathways. It also gave the results of an audit demonstrating that high quality equitable continence care was not reaching each patient. This article covers the literature search and the problems encountered in the setting up of a database and the development of a generic pathway, a symptom profile and specific pathways. It describes how each pathway evolved and was underpinned with the relevant evidence. It further describes the supporting information and design problems. Finally, it gives information on piloting the care pathways. PMID- 11868173 TI - NHS Direct: examining the challenges for nursing practice. AB - This article examines the impact of the launch of NHS Direct which was part of the Government's vision for a modern and dependable NHS. In particular, the challenges for nursing practice are detailed within a healthcare strategy which seeks to provide fast access primary health care to the whole population. The strengths and weaknesses of NHS Direct are discussed and its interface with other agencies explored. As NHS Direct will cover the whole of England by October 2000, the evidence to underpin its effectiveness must be considered in the light of media criticisms which indicate that the quality of advice given to callers is sometimes less than optimum. PMID- 11868174 TI - Biogel Reveal: a puncture indication system from Regent Medical. AB - Gloves provide an essential barrier against contamination and are an important item of personal protective equipment. Gloves used in clinical practice do leak although there is no direct evidence that such leaks result in transmission of infection. Double-gloving is recommended in theatre as a means of reducing hand injury during surgical procedures. Biogel Reveal is a double-glove puncture indication system that shows punctures as a visible green colour when damaged. PMID- 11868175 TI - The prison healthcare service: a workforce committed to change. AB - This article discusses the changes that are needed in the prison healthcare service, outlining the move to a multidisciplinary team approach, away from the outdated, medically dominated service which prevails today. This article supports the partnership arrangements put forward in 'The Future Organization of Prison Health Care' (Department of Health (DoH), 1999) and suggests how this can be achieved locally, calling on nurses working in the prison environment to be proactive and forge links with colleagues in local NHS services. The need for clinical governance to be firmly established and regular reports to be submitted to prison governors on the quality and provision of services is highlighted. PMID- 11868176 TI - Nurses are losing control of their profession. PMID- 11868177 TI - Dying and death are the final activities of living. PMID- 11868178 TI - Learning disability nurses must stay linked to the NHS. PMID- 11868179 TI - Discretionary points offer no incentive. PMID- 11868180 TI - Nurses must control their practice better. PMID- 11868181 TI - Clients are not being protected from abuse. PMID- 11868182 TI - Case 36: abuse of position. Speaking to patients and staff in an obscene and abusive way. PMID- 11868183 TI - Improving pain management: breaking down the invisible barrier. AB - There is compelling evidence that despite growing research into the complex neurophysiology of pain, the development of acute pain services, increasing educational interest in pain management and the proliferation of literature, many patients continue to suffer from unrelieved acute pain while in hospital. Educational efforts to bring about a change in practice have been relatively unsuccessful or slow to have real impact. Although it is still recognized that poor knowledge of pain control by all healthcare professionals is the major barrier to improving pain management, contemporary studies show that other, more subtle barriers can just as effectively inhibit a timely and effective response to patients' reports of pain. These barriers are not just the ones created by poor knowledge, myth and misconception; the most powerful barriers to change may be the invisible institutional barriers that can be entrenched within hospital policies and nursing rituals. PMID- 11868184 TI - Developing an oral assessment and intervention tool for older people: 3. AB - This is the last in a series of three articles which have examined oral care practices for older people. The first article (Vol 9(17): 1124-7) highlighted the importance of effective oral care. The second article (Vol 9(18): 2033-40) examined oral care interventions in detail to identify oral care that is evidence based. This article looks at the factors which have an impact on oral health, and which must be considered when assessing a patient's oral health status. The correlation between holistic assessment and careful documentation is examined, together with its associated relationship with maintaining effective oral care. The article also critiques a collection of published oral assessment tools to identify whether they would be appropriate for rehabilitation client groups. After identifying the lack of a suitable oral assessment tool within the literature, the need to develop an appropriate one is addressed. An oral assessment and intervention tool which has been developed in the author's clinical area is outlined. PMID- 11868185 TI - The needs of older people with learning disabilities. AB - There has been a growing realization that the needs of older people with learning disabilities are not being met. There has been little attention paid in nursing literature to the needs of this client group. This article will explore the issues about older people with learning disabilities and highlight some of the important areas that learning disability nurses may need to address. It is evident that there needs to be more collaboration between a range of different services and professionals. Areas of good practice can be used in developing innovative and flexible services, which are client led. Learning disability nurses may need to update their skills and knowledge and develop a 'critical practice' approach to meet the many challenges that older people with learning disabilities may pose. PMID- 11868186 TI - A model of independence for children with learning disabilities. AB - This article outlines the work undertaken to find a suitable model for planning respite care for children with disabilities. The model 'This is Me' and the informal life events assessment tool 'Hello, This is Me!', which supports it, are described. The use of a person-centred care planning system is well documented in the care of adults with learning disabilities (Sanderson, 1997). This process should begin at the start of a person's life, as recommended by The Children Act 1989, and the documents 'Signposts for Success' (NHS Executive, 1998) and 'Don't Forget Us' (Mental Health Foundation, 1997). The article describes the development of a person-centred approach to respite care which is underpinned with a basic care premise. PMID- 11868187 TI - Implementing 12-hour shifts on a cardiology nursing development unit. AB - This article, the first of two parts, discusses the implementation of 12-hour shifts using a locally devised nursing development unit (NDU) framework. A literature review and force-field analysis were undertaken to plan, implement and evaluate the introduction of the 12-hour system. The literature review identified five broad categories: effect on care delivery; nurse education; cost effectiveness; impact on staff; and implementation strategies. It also ascertained that the most successful methods of implementation were those that gained the cooperation of staff. The force-field analysis identified the restraining forces (e.g. tiredness, the European Working Time Directive and staff views) and therefore allowed the change agent to concentrate his limited time on combating these restraining forces. The second part of this series will examine the evaluation and audit of the 12-hour shift system. PMID- 11868188 TI - The new QUATTRO PLUS mattress replacement system. AB - Since the early development of alternating pressure air mattresses (APAMs) there have been regular alterations in design. These designs have led to improved reliability and performance. This article outlines the changes made by Talley Medical to the QUATTRO PLUS replacement mattress. Improved safety features and design modifications continue the process commenced many years ago which has seen the use of alternating pressure air mattresses become widespread throughout the UK. PMID- 11868190 TI - Cleanliness in hospitals is the role of nurses. PMID- 11868189 TI - Use of Pressurease and Airform mattresses in pressure ulcer care. AB - Large cell dynamic air mattresses are effective in the prevention of pressure ulcers but they still have inherent problems, e.g. the ability of the mattresses to move patients along the mattress through the inflation and deflation of the cells. Also, there have been reports that the inflation and deflation of cells can cause motion sickness. This article reviews two mattresses from Sareo Healthcare Ltd: a static system, Pressurease; and a unique dynamic system, Airform. These two systems offer a cost-effective and clinically effective method of pressure ulcer prevention and therapy. PMID- 11868191 TI - The use of hand gestures as self-generated cues for recall of verbally associated targets. AB - This study examined the effects of hand gestures as cues for recall of 40 previously described abstract and concrete words. Participants were either self cued (SC) with their own self-generated gestures, other-cued (OC) with someone else's gestures, or shown no cues (NC). The SC group had significantly better recall for both word types than either the OC or NC immediately and at a 2-week retrieval interval. Results also showed that when the SC group produced a meaningful gesture, concrete words were cued significantly more often than abstract words, but when total recall (cued and residual) was considered, abstract words were recalled equally well. These results are discussed in the context that hand gestures are a component of subjective organization and are thus distinctive cues for the producer that may facilitate or prime recall. PMID- 11868192 TI - One hundred years of laws in psychology. AB - Mainstream psychology in the 20th century has been conceived as a nomothetic science, but few psychological "laws" have been proposed. A PsycLit search of journal abstracts from 1900 to 1999 yielded a total of 3,093 "law" citations, or 22 per 10,000 entries, with two psychophysical laws (Weber's law and Stevens's power law) and two learning laws (Herrnstein's matching law and Thorndike's law of effect) as the most frequently cited. The number of law citations has been decreasing throughout the century, to 10 per 10,000 entries in the last decade, with few references to laws of recent origin. This could be the result of increasing doubts about the lawfulness of psychological processes coupled with a general preference for less ambitious terms (such as effects, principles, models, or functions). PMID- 11868193 TI - A naturalistic study of autobiographical memories evoked by olfactory and visual cues: testing the Proustian hypothesis. AB - The emotional and evocative qualities of autobiographical memories evoked by odors and visual cues were compared using a new repeated-measures paradigm in which the sensory cue was presented after the memory had been retrieved by its verbal label. Memory cues were chosen to be able to elicit salient memories. Results revealed that memories recalled in the context of odors were significantly more emotional than those recalled in the context of the same cue presented visually and by the verbal label for the cue. Odor-evoked memories also tended to make participants feel more "brought back" to the original event. This work is the first unequivocal demonstration that naturalistic memories evoked by odors are more emotional than memories evoked by other cues. PMID- 11868194 TI - The influence of naive causal theories on lay concepts of mental illness. AB - Two experiments, incorporating both real-life (Experiment 1) and artificial (Experiment 2) stimuli, demonstrated that lay concepts of mental disorders can be reliably predicted from subjects' naive causal theories about those disorders. Symptoms that are deeper causes (X, where X causes Y, which causes Z) are more important in lay concepts than intermediate causes (Y), which in turn are more important than terminal effects (Z). In addition, symptoms that cause or are caused by other symptoms are more important in lay concepts than symptoms not participating in any causal relationships. Implications of these results for current models of categorization and for research on lay theories of mental disorders are discussed, and future directions for research are suggested. PMID- 11868195 TI - The influence of contextual constraints on recall for words within sentences. AB - Two experiments examined predictions of the relational-distinctive processing view to account for the influence of sentence constraints on memory of target words. In Experiment 1, congruous expected words were recalled better than incongruous words. Words appearing in high-constraint sentences were recalled better than words appearing in low-constraint sentences. In Experiment 2, word expectancy and sentence constraint interacted so that unexpected congruous words appearing in high-constraint sentences were recalled better than their expected counterparts, but this difference was not present for low-constraint sentences. A hybrid model including aspects of the featural restriction model of sentence constraint and the relational-distinctive processing view is proposed. PMID- 11868196 TI - Working memory span: the effect of prior learning. AB - Recent work suggests that working memory span (WMS) tasks are not simple measures of the capacity to simultaneously store and process new information. Instead, these measures may be influenced by numerous factors, including proactive interference (PI). The current study examined whether WMS, like other memory tasks, is also influenced by PI from prior memory experiments. Experimentally experienced and naive participants completed a speaking span task. Span scores were lower for experienced than for naive participants, but other cognitive scores were not. In combination with other work, these results suggest that WMS estimates are not pure measures of capacity and may be partially determined by PI. PMID- 11868197 TI - Abiotrophia endocarditis: a case report and review of literature. AB - Nutritionally Variant Streptococci (NVS) were first grouped under viridans streptococci, although they differed from the latter by variant growth characteristics. NVS cause approximately 5% of cases of bacterial endocarditis. Infective endocarditis caused by NVS has a higher rate of complications than endocarditis caused by other viridans streptococci. Recently NVS were separated from other viridans streptococci to form a new genus Abiotrophia. Since then, only four case reports have described the clinical course of Abiotrophia endocarditis. Therefore, current knowledge on this disease derives from previous data on NVS endocarditis. We present the case of Abiotrophia endocarditis, followed by discussion of relevant literature. PMID- 11868199 TI - Unique nutrition support for research at the Cancer Research Center of Hawaii. PMID- 11868198 TI - School health education at the Queen Emma Clinics: a service-learning project at the John A. Burns School of Medicine. PMID- 11868200 TI - Palliative care--a service to improve the dying experience in hospitals and nursing homes. AB - The majority of Americans die in hospitals and nursing homes. There is substantial evidence in the medical literature, that care of the dying in such settings is often poor, and the obstacles to providing good end-of-life care remain numerous. Palliative care consultation has the potential to improve the care of patients dying in the institutional setting. PMID- 11868201 TI - Are people with Down's syndrome denied care? PMID- 11868203 TI - The RCN should exist for qualified nurses. PMID- 11868202 TI - Mixed blessings: is the NHS Plan revolutionary? PMID- 11868205 TI - Battle of the sexes still raging on the wards. PMID- 11868204 TI - Older people must be respected not abused. PMID- 11868206 TI - Case 35: nursing documentation. Inappropriate written remarks in patients' records. PMID- 11868207 TI - Indwelling catheters: practical guidelines for catheter blockage. AB - Catheterization is a medical decision and requires a team approach involving, where possible, the patient and carer, as well as those professionals administering care. Catheterization and subsequent effective catheter drainage is associated with many dangers and complications (Lowthian, 1998) and therefore the decision to catheterize someone should not be taken lightly. Catheterization should only be used in intractable incontinence when all other measures have been tried and proven to be ineffective. This article identifies some of the problems associated with the care of patients with indwelling catheters in the community setting, and examines the issues that can cause pain, distress and discomfort to patients. It looks at the various types of solutions to problems associated with catheterization. Case histories are used to illustrate the types of patients who may have been catheterized. It is hoped that nurses can learn from these examples, so that they will not only cope more effectively with these individuals, but also observe their own practice when initiating indwelling catheterization and developing care packages. PMID- 11868208 TI - Developing an activity measuring system in district nursing. AB - The current measurement of face-to-face contacts for calculating district nursing activity is inadequate for district nurses, their managers, and purchasers. With the growing emphasis on health outcomes, a measure is needed which demonstrates health gain and effectiveness, yet which does not place an additional burden for data collection on nurses' time. This article describes the preliminary findings of a project carried out in East Berkshire which suggest that such a measure is possible, and that effective care, including illness prevention, can be demonstrated. It shows that existing computer software can meet the demands for the measurement of health gain. While these are positive findings, district nurses, service managers and commissioners will need to allocate time to understand and interpret the data before drawing conclusions about the use of activity measurement in workforce planning, service costing and the effectiveness of district nursing interventions. PMID- 11868209 TI - From 'nits' to 'crabs'?: school nurses and sexual health. AB - The state of education and service provision relating to young people and sex, sexualities, and sexual health is currently a major national challenge. Nurses working in the compulsory education sector are at the direct interchange between clients and their obvious experiential and academic needs, and attitudes and systems that frequently seem to fail them. From concerned parents and members of the public, through the teaching and nursing professions, to the UK Government, the message is the same: something must be done to improve the current status quo. The problem lies in the different beliefs about what to do and how to do it. School nurses are in a prime position to improve sexual health education and services for young people. However, it is unreasonable to expect school nurses, who are frequently overburdened with task-oriented jobs, to be effective with an additional remit for sexual health education without adequate support, resources and effective professional training. This article explores the role of school nurses as it develops to encompass numerous aspects of sexual health care. In order to enhance and promote these changes in line with client needs the article examines the strengths, weaknesses, opportunities and threats (SWOT analysis) of this changing role. PMID- 11868210 TI - Pancreatic exocrine insufficiency and type 1 diabetes mellitus. AB - Studies in adults suggest that some patients with type 1 diabetes mellitus have pancreatic exocrine insufficiency (PEI). The primary aim of this study was to explore the association between pancreatic exocrine function and type 1 diabetes in young people under 17 years. The secondary aim was to evaluate the relationship between PEI in patients with diabetes, their clinical symptoms and blood glucose control. The importance of providing a highly trained multidisciplinary support network are also discussed. PMID- 11868211 TI - Developing an oral assessment and intervention tool for older people: 2. AB - This is the second of a series of three articles relating to oral care for older people. The first article highlighted the importance of effective oral care for older people (Vol 9(17): 1124-7). This article critically analyses a number of oral care agents and interventions to identify evidence-based practice relating to oral care for older people. Recommendations for practice are highlighted to guide practitioners in implementing safe and effective nursing interventions within oral care. The third article will critique a number of published oral assessment tools and describe an oral assessment and intervention tool that has been developed. PMID- 11868212 TI - The Flo-tech Adjuster chair from Medical Support Systems. AB - Pressure ulcer prevention in the seated patient is a neglected area. The early rehabilitation of bed-bound patients has reinforced the need for suitable seating to compliment pressure-reducing mattresses. Medical Support Systems has developed a hospital chair, the Flo-tech Adjuster, which addresses the problems of both pressure ulcer prevention and manual handling. The launch of the Flo-tech Adjuster has followed many years of research and constant refinement and improvement, and has filled a gap in the market for pressure-relieving devices. PMID- 11868213 TI - Reviewing the scope of professional practice. PMID- 11868214 TI - Humour on the job. PMID- 11868215 TI - Using community development approaches. PMID- 11868216 TI - Evaluation of mindfulness-based stress reduction intervention. PMID- 11868217 TI - Off-campus delivery of doctoral nursing education. PMID- 11868218 TI - A stitch in time: women and heart health quilt project. PMID- 11868219 TI - Refusal to care. PMID- 11868220 TI - Herbal products sparked symptoms. PMID- 11868221 TI - Nursing with heart. PMID- 11868223 TI - 1st Canadian conference on hepatitis C. PMID- 11868222 TI - Adolescent care in religious settings. PMID- 11868224 TI - Branching out. The dissemination and use of nursing research. PMID- 11868226 TI - Introducing critical care nursing at an undergraduate level. PMID- 11868225 TI - Reflections on using Parse's theory of human becoming in a palliative care setting in Pakistan. PMID- 11868227 TI - How valuable is peer review? PMID- 11868228 TI - Motivational interviewing. PMID- 11868229 TI - Memory mechanisms in pigeons: evidence of base-rate neglect. AB - In delayed matching to sample, once acquired, pigeons presumably choose comparisons according to their memory for (the strength of) the sample. When memory for the sample is sufficiently weak, comparison choice should depend on the history of reinforcement associated with each of the comparison stimuli. In the present research, pigeons acquired two matching tasks in which Sample S1 was associated with one comparison from each task, C1 and C3, whereas Sample S2 was associated with Comparison C2, and Sample S3 was associated with Comparison C4. As the retention interval increased, the pigeons showed a bias to choose the comparison (C1 or C3) associated with the more frequently occurring sample (S1). Thus, pigeons were sensitive also to the (irrelevant) likelihood that each of the samples was presented. The results suggest that pigeons may allow their reference memory for the overall sample frequency to influence comparison choice, independent of the comparison stimuli present. PMID- 11868230 TI - Unawareness is more than a chance event: comment on Lovibond and Shanks (2002). AB - P. F. Lovibond and D. R. Shanks (2002) suggested that expectancy of the unconditional stimulus and emotional ratings are valid indexes of awareness in Pavlovian conditioning and that participants are aware if they can discriminate the conditional stimuli. However, research suggests that processes that are irrelevant to awareness affect these measures. Further, as awareness refers to conscious experience, a valid measures needs to index subjective state rather than discrimination ability. In support, research using subjective measures has demonstrated qualitatively different effects depending on whether participants reported being aware or unaware of the stimuli. In this research, participants reported being unaware of the stimuli even though they were clearly able to discriminate the stimuli. These findings question the validity of Lovibond and Shanks' concept of awareness and their suggestion of a close association between conditioning and awareness. PMID- 11868231 TI - The role of awareness in Pavlovian conditioning: empirical evidence and theoretical implications. AB - This article reviews research over the past decade concerning the relationship between Pavlovian conditioning and conscious awareness. The review covers autonomic conditioning, conditioning with subliminal stimuli, eyeblink conditioning, conditioning in amnesia, evaluative conditioning, and conditioning under anesthesia. The bulk of the evidence is consistent with the position that awareness is necessary but not sufficient for conditioned performance, although studies suggestive of conditioning without awareness are identified as worthy of further investigation. Many studies have used inadequate measures of awareness, and strategies for increasing validity and sensitivity are discussed. It is concluded that conditioning may depend on the operation of a propositional system associated with consciousness rather than a separate, lower level system. PMID- 11868232 TI - Standard delay eyeblink classical conditioning is independent of awareness. AB - P. F. Lovibond and D. R. Shanks (2002) suggested that all forms of classical conditioning depend on awareness of the stimulus contingencies. This article considers the available data for eyeblink classical conditioning, including data from 2 studies (R. E. Clark, J. R. Manns, & L. R. Squire, 2001; J. R. Manns, R. E. Clark, & L. R. Squire, 2001) that were completed too recently to have been considered in their review. In addition, in response to questions raised by P. F. Lovibond and D. R. Shanks, 2 new analyses of data are presented from studies published previously. The available data from humans and experimental animals provide strong evidence that delay eyeblink classical conditioning (but not trace eyeblink classical conditioning) can be acquired and retained independently of the forebrain and independently of awareness. This conclusion applies to standard conditioning paradigms; for example, to single-cue delay conditioning when a tone is used as the conditioned stimulus (CS) and to differential delay conditioning when the positive and negative conditioned stimuli (CS+ and CS-) are a tone and white noise. PMID- 11868234 TI - What is learned in sequential learning? An associative model of reward magnitude serial-pattern learning. AB - A computational model of sequence learning is described that is based on pairwise associations and generalization. Simulations by the model predicted that rats should learn a long monotonic pattern of food quantities better than a nonmonotonic pattern, as predicted by rule-learning theory, and that they should learn a short nonmonotonic pattern with highly discriminable elements better than 1 with less discriminable elements, as predicted by interitem association theory. In 2 other studies, the model also simulated behavioral "rule generalization," "extrapolation," and associative transfer data motivated by both rule-learning and associative perspectives. Although these simulations do not rule out the possibility that rats can use rule induction to learn serial patterns, they show that a simple associative model can account for the classical behavioral studies implicating rule learning in reward magnitude serial-pattern learning. PMID- 11868233 TI - Autonomic and eyeblink conditioning are closely related to contingency awareness: reply to Wiens and Ohman (2002) and Manns et al. (2002). AB - S. Wiens and A. Ohman (2002) disputed the conclusion that Pavlovian conditioning is strongly related to contingency awareness (P. F. Lovibond & D. R. Shanks, 2002) on the basis that an inappropriate definition of awareness was used. J. R. Manns, R. E. Clark, and L. R. Squire (2002) contended that delay eyeblink conditioning is independent of awareness. The authors of the present article consider these arguments, highlight several problems in the new studies described by the commentators, and conclude that there is still little evidence for unconscious conditioning in either subliminal autonomic conditioning or eyeblink conditioning. The most parsimonious account of existing data is that a single learning process gives rise to both awareness and conditioned responding. Further progress in evaluating the possibility of unconscious conditioning would be facilitated by the development of more completely specified and testable dual process models. PMID- 11868235 TI - Second-order contrast based on the expectation of effort and reinforcement. AB - Pigeons prefer signals for reinforcement that require greater effort (or time) to obtain over those that require less effort to obtain (T. S. Clement, J. Feltus, D. H. Kaiser, & T. R. Zentall, 2000). Preference was attributed to contrast (or to the relatively greater improvement in conditions) produced by the appearance of the signal when it was preceded by greater effort. In Experiment 1, the authors of the present study demonstrated that the expectation of greater effort was sufficient to produce such a preference (a second-order contrast effect). In Experiments 2 and 3, low versus high probability of reinforcement was substituted for high versus low effort, respectively, with similar results. In Experiment 3, the authors found that the stimulus preference could be attributed to positive contrast (when the discriminative stimuli represented an improvement in the probability of reinforcement) and perhaps also negative contrast (when the discriminative stimuli represented reduction in the probability of reinforcement). PMID- 11868236 TI - Amphetamine and morphine produce a conditioned taste and place preference in the house musk shrew (Suncus murinus). AB - Rats have been shown to avoid consuming a flavor, but prefer a location, previously paired with amphetamine or morphine. A series of 4 experiments evaluated the hedonic properties of amphetamine and morphine in the house musk shrew (Suncus murinus), an insectivore that (unlike rats) is capable of vomiting when exposed to toxins. Unlike rats, amphetamine (20 mg/kg) and morphine (20 mg/kg) produced both a conditioned sucrose (0.3 M) and saccharin (0.1%) preference in shrews (administered intraperitoneally), when measured by both a 1- and a 2-bottle test. At the same dose, both drugs also produced a place preference in shrews. These results suggest that the potential of rewarding drugs to produce taste avoidance may vary on the basis of the ability of the species to vomit. PMID- 11868237 TI - Transfer across unconditioned stimuli in serial feature discrimination training. AB - The transfer of conditioned modulation across conditioned stimuli (CS) and unconditioned stimuli (US) was examined in 3 experiments that used Pavlovian appetitive training procedures with rats. In Experiment 1, after training in a positive patterning discrimination (X-->A+/X-/A-), X increased conditioned responding elicited by another trained-then-extinguished CS as long as that CS had been trained with the same US as was used in discrimination training. In Experiment 2, after training with a feature-negative discrimination (X-->A-/A+), X inhibited conditioned responding elicited by another trained-then-extinguished CS as long as that CS had been trained with the same US. Experiments 1 and 2 used a between-groups design, whereas Experiment 3 used a more powerful within-groups design. In Experiment 3, rats were trained in a feature-positive discrimination (X-->A+/A-). In transfer tests, X increased conditioned responding elicited by another CS trained then extinguished with the same US from training. This increase was greater than the X increased conditioned responding elicited by another CS trained then extinguished with a different US from training. The results supported the suggestion that features trained in serial discrimination tasks influence behavior indirectly by transiently raising or lowering the threshold for activation of the US representations by its target stimuli and by any other stimuli that may be associated with that US. Other interpretations of the findings were also considered. PMID- 11868238 TI - Reinstatement of fear to an extinguished conditioned stimulus: two roles for context. AB - The authors studied the role of context in reinstatement. Freezing was reinstated when the conditioned stimulus (CS) was extinguished in 1 context and rats moved to another context for reexposure to the shock unconditioned stimulus (US) and test. It was also reinstated (rather than renewed) when rats were shocked in the extinction context and moved to another context for test. This reinstatement was CS specific and reduced by nonreinforced exposures to the extinction context. Rats shocked in the context in which a stimulus had been preexposed froze when tested in another context. These findings suggest 2 roles for context in reinstatement: conditioning of the test context (M. E. Bouton, 1993) and mediated conditioning by the extinction context (P. C. Holland, 1990). PMID- 11868239 TI - [Pituitary function after transsphenoidal surgery for pituitary adenoma]. AB - Nowadays transsphenoidal surgery is the method of choice in most cases of pituitary adenoma treatment, both functioning and non-functioning. This method is considered to be safer than transcranial approach, as lower incidence of complications, particularly hypopituitarism, is observed. The aim of the study was to evaluate the pituitary function after transsphenoidal surgery. 20 patients (mean age 49.3 +/- 13.9 years) were included into the study. The anterior pituitary insufficiency was defined as an inadequate excretory response to metopirone, LH-RH and TRH stimulation. Diabetes insipidus was diagnosed based on clinical symptoms. Hypopituitarism was diagnosed after surgery in 7 patients, 3 cases had adrenal and gonadal insufficiency, 1 patient had insufficiency of the pituitary-thyroid and gonadal axis and 3 subjects panhypopituitarism. Diabetes insipidus was still present in 2 patients, 3 months after surgery. We conclude that transsphenoidal approach in pituitary adenoma surgery is connected with low risk of iatrogenic hypopituitarism. PMID- 11868240 TI - [Nutritional status of patients with functioning graft assessed by clinical examination, anthropometry and bioimpedance]. AB - Malnutrition is very frequent comorbid factor in chronic renal failure and its prevalence both in the predialysis period as well as on maintenance dialysis is high. The aim of the study was to assess the nutritional status in patients after successful kidney transplantation. 109 patients (47 F, 67 M) of mean age 39.9 +/- 11.5 years were analyzed. Mean time after transplantation surgery was 32.2 +/- 37 months and the maintenance dialysis treatment period prior to transplantation- 28.4 +/- 22 months. Nutritional status was assessed with clinical examination based on the SGA scale, anthropometric measurements as well as body composition estimation with bioimpedance. Daily food intake was also monitored with three-day dietary questionnaire. All above analyses were also performed in 25 healthy control subjects with corresponding sex and age distribution. No differences between all analyzed bioimpedance and anthropometry parameters were found between studied patients and controls. 79% of patients were classified as well nourished, 20%--as mildly or moderately malnourished and only 1%--as severely malnourished according to SGA scale. The BMI values less than 21 kg/m2, i.e. suggesting malnutrition were found in 23.3% of patients, whereas values above 25 kg/m2, i.e. suggesting overweight or obesity--in almost 40%. Interestingly, as high as 82.5% of studied patients were characterized by significant weight gain since last "dry weight" assessment on maintenance dialysis up to the time of study (by mean 9.42 +/- 6.9 kg). Obtained results permit us to conclude, that the prevalence of nutritional status abnormalities are relatively frequent among patients with functioning graft. Malnutrition can be demonstrated in more than 20% of the study population, which should be considered however to be markedly lower as compared to most reports regarding dialysis populations. Weight gain during posttransplant period as compared to maintenance dialysis is marked and common; thus the prevalence of obesity is also quite common and reaches 40% of tested patients. PMID- 11868241 TI - [Effect of vitamin E-modified membrane on expression of coreceptors CD4, CD8 on lymphocytes in chronic hemodialysis patients]. AB - Chronic renal failure induces a clinical state of cellular and humoral immunodeficiency that also depends on the time duration of blood contact with the wide spectrum of dialysis membranes use during long-term hemodialysis treatments. In end stage renal failure (ESRD) patients it is possible to induct state of chronic inflammation mostly caused by leukocytes and complement activation. It is postulated that the vitamin E-coated dialysis membrane minimalizes unbiocompatible reactions that generate smaller amounts of reactive oxygen species (ROS). The purpose of this study was to analyze the effect of classical and vitamin E coated cellulose membranes on the expression of CD 4 and CD 8 adhesion molecules on lymphocytes during HD in 10 patients using flow cytometric analysis. The study protocol included the measurement of molecules expression using cellulose membrane (Clirans RS15, TERUMO Corp., Japan), and the same membrane coated by vitamin E (Excebrane, Clirans E15, TERUMO Corp., Japan) during 20 dialysis sessions with each kind of membrane. During dialysis with classical cellulose membrane, significant decrease of lymphocyte serum level and increase of lymphocyte CD4 expression was observed. During the session with vitamin E coated membranes we did not observe any significant changes in serum CD4, CD8, CD4+8+ lymphocyte level, and also lymphocyte CD4, and CD8 expression on lymphocytes. Our findings suggest the potential role of vitamin E-coated cellulose membrane to minimalize negative reaction of the T lymphocyte subpopulation in ESRD patients treated on long-term dialysis. PMID- 11868242 TI - [Pathological parameters of endometrial cancer and presence of metastases in pelvic lymph nodes]. AB - Endometrial cancer has become a more frequent neoplasm of the female genital tract. The role of lymphadenectomy in surgical treatment of this neoplasm has not been finally defined. The aim of the study was to determine relationship between pathological parameters of endometrial cancer and presence of metastases in pelvic lymph nodes. Forty one patients with endometrial cancer were treated by extended hysterectomy with pelvic lymphadenectomy. The precise Fisher test and logistic regression test were applied in the analysis of relationship. An intrinsic connection between presence of metastases in pelvic lymph nodes and cancer grade, depth of myometrium infiltration depth and infiltration of vascular spaces was found. On the other hand, histological type of neoplasm and characteristic of its growth does not seem to have connection with presence of metastases in pelvic lymph nodes. Pelvic lymphadenectomy seems to give profound information of of process advancement and indications for supplementary treatment. PMID- 11868243 TI - [Etiological spectrum of symptomatic epilepsy in adults]. AB - The purpose of this study was to establish the frequency of incidence of epileptic seizures among adult patients of the Neurological Department. The second aim was to establish the causes of the symptomatic epilepsy. The material comprised of 205 patients, who were hospitalized because of first epileptic seizures. The etiology of epilepsy was based on history, neurological findings and diagnostic investigations. Only the patients over 20 years old were included in the study. The analysed group of 205 cases was 6.6% of all hospitalized patients during a period 5 years. The average age of these patients was 57.5; 58% -were male, 42%--female. Vascular lesions of the brain were the most frequent cause of epilepsy (46.4%); alcohol abuse--13.6%, brain tumor--10.2%, head trauma- 8.8%, alcohol and head trauma--6.3%. Inflammation, degeneration and metabolic processes were very rare. In 8.8% of cases with late epilepsy, the etiology of disease remained unknown. In the group with vascular lesions of the brain, ischaemic stroke was the most frequent (62.8%). Among the patients with ischaemic stroke females prevailed. Alcohol-associated seizures were disorders very often among males. The literature analysis and our own notices showed that the frequency of symptomatic epilepsy in the elderly population increased. The connection between this observations and the higher average survival and the frequency of vascular diseases of the brain, is possible. The etiology of epilepsy in very elderly people is not clear and probably is associated with several factors. This group of patients comprised 25.5% of all our material. PMID- 11868244 TI - [Comparative testing of aspirin esterase and cholinesterase activities in sera of healthy blood donors]. AB - Activities of aspirin esterase, EC 3.1.1.55 and cholinesterase, EC 3.1.1.8 were measured in sera of 106 healthy blood donors. Surprisingly high correlation of these two activities has been observed. Aspirin esterase exhibited about 10 times lower activity than cholinesterase, when expressed in mkat/L. PMID- 11868245 TI - [Long term outcome in patients with and without restenosis after coronary angioplasty]. AB - Restenosis at dilatation site still constitutes a significant limitation of percutaneous coronary revascularisation. Majority of patients with restenosis are treated with repeat angioplasty, although its impact on long-term outcome is still little known > Very few studies focused on this issue bring rather discrepant results. The present study is aimed at assessing the impact of restenosis on long-term outcome in patients treated with coronary angioplasty. A group of 567 patients, who in the years 1987-1996 had successfully undergone percutaneous balloon coronary angio-plasty (PTCA) at our Clinic, was retrospectively divided into two groups: a group comprising 188 patients (33.2%) suffering from recurrent angina in whom restenosis had been established through control angiography, and a group comprising 379 patients (66.8%) who during the observation period exhibited no angina symptoms, or in whom the control angiography did not reveal restenosis. The restenotic patients were older (p = 0.007), more frequently exhibited symptoms of unstable angina upon PTCA (p < 0.0001), and there were also fewer smokers among them (p = 0.02). Furthermore, restenotic patients more frequently had multivessel and multilesion angioplasty (p = 0.025; p = 0.004, respectively). Restenosis after the first PTCA was treated by repeated angioplasty in 149 (79.3%) patients, 26 (13.8%) underwent CABG operation and 13 (6.9%) patients were treated pharmacologically without repeated revascularization. Within the 5-year observation period the mortality rate in both groups did not differ significantly (5.9% vs. 4.0%). Restenotic patients sustained myocardial infarctions more frequently (8.0% vs. 3.2%, p = 0.01), had significant atherosclerosis progression (37.2% vs. 15.0%, p < 0.0001), and were more frequently subjected to repeated interventions (37.2% vs. 15.0%, p < 0.0001), both PTCA (79.3% vs. 11.6% p < 0.0001) and CABG surgery (32.5% vs. 4.2%, p < 0.0001), as well as to multiple re-interventions (31.9% vs. 4.8%, p < 0.0001), in comparison with the non-restenotic ones. Analysis of functional status of patients assessed in compliance with the CCS criteria at the end of the observation period proved that significantly more non-restenotic patients did not experience angina, or experienced it rather sporadically (CCS Class 0 and 1). Logistic regression uni- and multivariate analysis proved that restenosis is an independent risk factor of myocardial infarction, reinterventions--also multiple ones--and cardiac events, although not mortality. PMID- 11868246 TI - [Self-estimation of the quality of life in adult patients after surgical correction of atrial septal defect type II (ASD II)]. AB - Health estimation was performed in 134 patients (where 67% were women), aged 17 70, mean 42 years, 2-3 years after surgical correction of atrial septal defect type II (ASD II). The study consists of clinical examination and self-estimation of the quality of life with help of a mall questionnaire, with return ratio of 90%. The improvement of health status was declared by 80% of patients, where 23% stated considerable improvement. While 15% did not confirm any significant changes and 5% noticed worsening quality of life status (mainly connected with postoperative pain). The physical condition improved similarly, with range of tolerable physical effort doubled. The frequency of dyspnea, chest pain and palpitation decreased from 72%, 67% and 87% to 47%, 43% and 47%, respectively, as well as their intensity. More over, the frequency of anxiety decreased from 70% to 62% with reduction of its intensity. Both, before and after surgery, the environmental estimation and self-estimation was very good (77% versus 78%, 78% versus 89%) respectively, and predominant were optimistic attitudes. Post operative improvement of the quality of life correlating to the clinical state, confirms the suitableness of surgical correction of ASD II, independent of age. PMID- 11868247 TI - [Hyperlipidemia as a risk factor for coronary heart disease among elderly people]. AB - The significance of hyperlipidemia in risk prognosis of coronary heart disease had been well documented in cases of young and middle age people. There is no explicit agreement of how important of a role this risk factor plays in predicting a threat of coronary heart disease in elderly people. The role of hyperlipidemia as a potential risk factor of cardiovascular disease in cases of elderly people, is still from a scientific point of view an interesting and current subject. The confirmation of existence of such risk factors could help make proper prophylaxis therapy possible and place emphasis on the necessity to initiate a hypolipemic therapy in case of elderly people concerning primary and secondary prevention. The review of bibliography concerning this subject and presented by different authors shows that existing significant differences concerning this problem. PMID- 11868248 TI - [Post-transplant nephropathy and arterial hypertension]. AB - The introduction of new immunosuppressive regimens results in the significant improvement in the outcome of patients after kidney transplantation. However, about 5 percent of renal transplants are lost every year. Not only immunological (alloantigendependent) but also nonimmunological (alloantigen-independent) factors are involved in late graft loss. Among them, hypertension, hyperlipidemia, proteinuria, genetic predisposition, viral infection and nephrotoxicity of immunosuppressive drugs contribute to the development and to the progression of chronic post-transplant nephropathy. Hypertension can be both the cause and the consequence of chronic allograft failure. Hypertension is frequently observed before transplantation, persists after grafting and increases the risk of chronic allograft nephropathy. Hypercholesterolemia, obesity, atheromatosis, polycythemia, and excessive salt intake are factors contributing in post-transplant hypertension. However, in some cases, hypertension can be transferred with the grafted kidney, as observed in normotensive patients before renal transplantation. In 1 to 12 percent of cases, the cause of post transplant hypertension is the stenosis of the transplant artery. Sometimes the presence of hypertension in renal recipients may result from the recurrence of glomerulonephritis or from the development of glomerulonephritis de novo in the graft. Also immunosuppressive treatment with corticosteroids and cyclosporine A contributes to the increased prevalence of hypertension by 20-30 percent. The development of the graft nephroarteriolosclerosis as a consequence of hypertension accelerates the progression of the post-transplant nephropathy. Adequate control of the arterial pressure (< 140/90) should be achieved in all renal transplant recipients. Reduction in protein and salt intake is important to reduce hyper-filtration and slows the progression of transplant nephropathy. However, pharmacological treatment is usually needed. Angiotensin-converting enzyme inhibitors, angiotensin II type I receptor antagonists exhibit beneficial hemodynamic effect leading to the reduction of glomerular hypertension and proteinuria. Calcium antagonists besides their systemic antihypertensive effect, can protect renal grafts from vascular and renal toxicity of CyA. Sometimes, combined therapy with these and other antihypertensive drugs and diuretics is necessary. PMID- 11868249 TI - [Histocompatibility HLA system of man. Considerations in the light of current concepts. IV. Soluble HLA class I antigens (sHLA-I)]. AB - Soluble class I human leukocyte antigens (sHLA) have been detected in serum, sweat, lymphatic fluid, urine and cerebrospinal fluid. The levels vary among different individuals and are significantly affected by inflammatory diseases and organ rejection. This article discusses the clinical significance of levels of serum HLA class I antigens, both in patients with viral diseases and following organ transplantation, as well as the potential involvement of such antigens in the immune response. The potential use of sHLA in clinical medicine is far reaching. sHLA-peptide complexes may find wide application, particularly for the treatment recipients. Although further in vivo studies are required, available data show the efficacy of sHLA to regulate T-cell function. PMID- 11868250 TI - [Histocompatibility HLA system of man. Considerations in the light of current concepts. V. sHLA-I in patient's body fluids]. AB - The retrospective estimation of concentrations of soluble HLA class I antigens (sHLA-I) in blood serum and in other body fluids of patients was executed. The physiological concentration of sHLA-I is significantly upregulated in various diseases and during inflammation, and following organ transplantation. This suggested that sHLA-I might serve as a marker of pathological changes. Observed differences in sHLA-I levels among different diseases could reflect variability in genetic factors, pathophysiology or disease activity. The authors show on usefulness of obtained results to comparative analysis of particular HLA class I allospecificities concentrations characteristic for some diseases, for example HLA-Cw7 in patients with SNHL, or HLA-B27 in zzsk. Estimation of sHLA-I concentrations in blood sera of patients could represents a good diagnostic and prognostic marker in monitoring of course disease and its activity. PMID- 11868251 TI - [Emotional disorders in patients with multiple sclerosis]. AB - Multiple sclerosis (MS) is the most common demyelinating disease of the central nervous system, especially young adults. Although MS is usually looked on as a disorder of the sensory and motor systems it can also be associated with emotional dysfunctions and changes in personality. The depression, bipolar disorders, euphoria, and pathological laughing and crying are most frequently associated with the disease. Authors present a review of current opinions on pathogenesis, diagnostic criteria and treatment of emotional problems in MS patients. PMID- 11868252 TI - [Bronchial carcinoid in a 7-year-old boy]. AB - A case of a 7-year-old boy admitted to the clinic with severe symptoms of 1-month lasting pneumonia not responding to antibiotics is presented. The chest X-ray confirmed inflammatory process in left lung parenchyma. Due to unsuccessful further preservative treatment, bronchoscopy and CT of the thorax were performed. They showed the presence of a tumor narrowing the left main bronchus. Histopathologic examination of the tissue taken during bronchoscopy revealed carcinoid. Through the left-sided thoracotomy, the resection of a 5 cm large oval shaped tumor, as well as the distal part of the left main bronchus was done. During clinical observation the child did not present any symptoms of the carcinoid syndrome. Urine levels of acids: homovanillic, vanillylmandelic and 5 hydroxyindolylacetic were normal. Bronchoscopy and X-ray of the chest directly, two weeks after and six months after resection were normal. Childhood primary pulmonary neoplasms are rare and the most frequent malignant tumors are bronchial adenomas. About 80-90% of them are carcinoids. The period from initial symptoms to clinical diagnosis and the institution of treatment, usually lasts several months. In our case it was a seven-week-long period. When cough, weezing, hemoptysis and inflammation of lung parenchyma are prolonged, carcinoid should always be considered in differential diagnosis. Radiological changes are usually nonspecific in cases of bronchial adenomas. Bronchoscopy with biopsy and CT scan are investigations that are decisive. Because of relatively low malignancy of carcinoid, results of the surgical treatment are good even in presence of metastases in regional lymphatic nodules. PMID- 11868253 TI - [Thyroglossal duct cyst imitating mediastinal tumor]. AB - A rare case of thyroglossal duct cyst mimicking the mediastinal tumour is described. Due to localisation, simple cystectomy was performed. The early and two year postoperative results are good, any recurrence of the cyst has not been observed. PMID- 11868254 TI - [Distribution of serotonin and its metabolites in the brain structures and immunosuppression in submissive mice]. AB - The development of submissive behaviour in C57BL/6J mice in the sensory contact model was associated with an increase in the content of serotonin (5-HT) in the amygdala, hippocampus, dopaminergic nuclei A11, A10, A9, as well as in the caudate nucleus and hypothalamus after 10 and 20 days of confrontations compared to the controls. The level of 5-HT metabolite 5-hydroxyindolacetic acid (5-HIAA) was significantly higher in the most structures examined after 20 daily encounters as compared to animals with experience of 10 confrontations. The time course of submission over 10 or 20 days resulted in an increase of 5-HIAA/5-HT ratio in the midbrain nucleus raphe, nucleus accumbens, A9 and hypothalamus. In mice immunised on the 10th or 20th day of confrontations, the immune response inhibition was observed while its level remained unchanged after more prolonged confrontations (40 days). Thus, the experience of defeats during 10 days shown to be accompanied with an activation of 5-HT system in a number of the brain structures, produced immunosuppression. With increasing number of confrontations the ratio 5-HIAA/5-HT was decreased in the same structures and a tendency to the immune response elevation appeared. PMID- 11868255 TI - [Recording of dependence of O2 consumption in the rat skeletal muscle on O2 tension in the incubation medium]. PMID- 11868256 TI - [Activation of the human cortex during visual attention and selection]. AB - Cortical activation in visual discrimination tasks was estimated by measurement of the CNV (contingent negative variation) and N1-P3 components of visual ERPs in frontal, parietal, occipital and temporal leads recorded in 18 young healthy adults. In all investigated tasks, the maximal values of CNV and ERPa were observed in parietal regions. The estimation of cortical readiness state (CNV) is quite a useful procedure in the attention tasks because amplitude and stability of ERPs depend on preceding cortical excitability. The prevalence of parietal activation in visual attention tasks may be considered as the dominance of occipito-parietal way (stream) in human visual attention system. PMID- 11868257 TI - [Selectivity of opioid peptide effects on excitability and various sensory inputs in LPl1 and PPl1 command neurons participating in defensive behavior of the snail Helix lucorum]. AB - Opioid peptides effects on neural membrane as well as neural responses evoked by sensory stimuli with different modality and site of application, were investigated in L-RPII command neurones of defensive behaviour of semi-intact preparation in the land snail Helix lucorum. Met-enkephalin (10 uM) application onto the snail CNS increases membrane excitability and produces facilitation of neural responses evoked by quinine solution (0.5%) application onto snail head and depression of reactions evoked by tactile stimulation of the head. Met enkephalin in dose of 0.1 uM initiates only a depression of neural responses evoked by tactile stimulation of the head. Leu-enkephalin (10 uM) application suppresses neural reactions evoked by tactile stimulation of the head. Membrane excitability and neural responses evoked by quinine application onto the snail head do not change after leu-enkephalin administration. Effects appear 10-20 min after initiation of the peptide application. Initial neural responses were observed 15-30 min after CNS washing with Ringer solution. In addition, facilitation of neural responses evoked by chemical stimulation of the snail head was found 30-50 min after leu-enkephalin washing. Peptides do not change neural responses evoked by tactile stimulation of the snail foot. Neural effects of peptides were prevented by simultaneous naloxon administration (50 uM). Experimental results show selective opioid peptides' effects on excitability and plasticity of L-RPII neural inputs with site- and modality-specifics. PMID- 11868258 TI - [Comparison in changes of glutamate level in the rat nucleus accumbens induced by D1- and D2-dopamine receptors during feeding]. AB - The influence of dopamine D1- and D2-like receptors blockage on glutamate level in the n. accumbens of Sprague-Dawly rats during feeding was investigated by in vivo microdialysis combined with HPLC-EC analysis. Food intake resulted in a decrease in extracellular glutamate level. Infusion of D1-like dopamine receptor blocker (SCH-23390, 0.01 mM) into the n. accumbens did not change this effect. Infusion of D2-like dopamine receptor-blocker (raclopride, 0.1 mM) into the n. accumbens caused an increase in extracellular glutamate level during feeding. The findings suggest, that decrease in extracellular glutamate level in n. accumbens is caused by dopamine D2-like, but not D1-like receptors activation. PMID- 11868259 TI - [Effect of sodium nitroprusside on erythrocyte phagocytosis by peritoneal macrophages in white rats]. AB - The study investigates into the role of exogenous nitric oxide (NO) in erythrophagocytosis in rats in vitro. The data indicate that NO enhances the ability of macrophages to adhere and ingest erythrocytes with the rise of nitric oxide concentration in cultural medium. NO influence on red blood cells has been shown to be more significant than its effects on macrophages. The reaction of NO with macrophages results in promotion of initial stages of erythrophagocytosis: macrophages' ability of adhering to the plate and to adhere erythrocytes increases. However, the final stages of erythrophagocytosis are activated just by the influence of NO on red blood cells. Therefore our results have demonstrated that the target point for NO is erythrocyte. We have also confirmed that NO effects are dose-dependent. PMID- 11868260 TI - [Effect of hypervolemia on the adrenergic reactivity of the arterial system]. AB - In anaesthetised rats, dependence of haemodynamics upon increase in the blood volume due to infusion of polyglucin in amounts 1.2 and 2.4 ml (8% and 16% of the blood volume in rats, respectively) < was studied. In the former case the initial blood pressure increased by 30%, in the latter case--by 42%; cardiac output--by 16% and 40%. The metasone pressor effects, at the were reliably decreased by 27% and 65%, and those of general peripheral resistance--by 40% and 80, respectively. The cardiac output changes did not differ significantly. The data obtained suggest an effect of the blood volume increase upon a drop of the arterial system's adrenoreactivity as a result of increase in initial blood pressure. PMID- 11868261 TI - [Genoprotective human blood activity and its mechanisms]. AB - Anticlastogenic properties of plasma and proteins (albumin and gamma-globulin) of the human blood were studied using seeds of Crepis capillaris (chromosome aberration assay). Antimutagen p-amino-benzoic acid was used as a comparative reagent. Anticlastogenic activity dependent of processing conditions of the biosubstrate used; for the pre-processing and combined processing anticlastogenic effect was higher than for post-processing, the processing properties of the blood being higher than those of the blood proteins. Anticlastogenic potential of biosubstrates did not depend on mutation inductor. Complex-formimg properties of plasma and blood albumen have been revealed using spectrop-hotometry through the substantial spectral displacement--relative to the expected spectrum--for the mixture of biosubstrata and mutagens. Using chemoluminescence, all plasma, albumin and gamma-globulin concentrations have been shown to enhance generation of hydroxyl radical of the Fenton reagent, especially for albumin in 1.0 g/l concentration. The general trend for all experiments was that the said substances diminished the stimulating effect as their concentrations grew. Peroxidation of yolk lipoproteids showed that only high concentrations of blood's plasma and albumen have antioxidizing properties. gamma-Globulin did not reveal any ability to inhibit lipid peroxidation of yolk lipoproteids. Complex-forming mechanisms of blood's albumen and antioxidizing property of human plasma and proteins have been proved to form the blood's anticlastogenic potential. PMID- 11868262 TI - [Age and sex characteristics of phospholipid composition of Wistar rat erythrocytes during postnatal ontogenesis]. AB - The composition of phospholipids and fatty acids of erythrocyte lipids of intact mature Wistar rats and their progeny in the age of 10, 15, 20, 30, 40, 60, 90 and 110 days was studied with the methods of micro-thin-layer and gas-liquid chromatography. Sex features and age mechanism of erythrocyte lipids composition forming in physiologically standard conditions were established. The fact that the composition of erythrocyte lipids of the untreated grown-up male rats in comparison with the female species is characterised with a lower concentration of phosphatidylinositol, a higher level of phosphatidylcholine and sphingomyelin, a higher saturation level of molecular types of phospholipids, was described. Three age periods of homeostasis of the erythrocyte lipids of the intact rats were distinguished. The postembryonic period (30-40 days after birth) is characterised with forming of the basic pool of erythrocyte lipids: getting to the stability in levels of the basic components of phospholipids and their fatty acids, coming out of phosphatidylinositol by the 40th day after birth. The period of sex-growing (60 days) is accompanied by spasmodic changes in the levels of fatty acids which is most common for female species: rapid increase in saturated and omega 3 polyunsaturated fatty acids, decrease in monoene and omega 6 polyene fatty acids. The period of maturity (110 days) is characterised by a slow metabolism of structural lipids and a stabilisation of their composition. PMID- 11868263 TI - [Changes in metabolic activity of neurons in the anterior hypothalamus nuclei during hyperthermia, fever, and hypothermia]. AB - Differently directed changes in metabolic activity of anterior hypothalamic nuclei's neurons in rats during hyperthermia, fever, and hypothermia were revealed with histochemical methods. During hyperthermia, the activity of energy metabolism enzymes increased as well as RNA content in the neurons of supraoptic, paraventricular and median preoptic anterior hypothalamic nuclei. This is shown by an increase in the metabolic activity of neurons of these nuclei. Metabolic activity in neurons of median preoptic nuclei decreased and was not changed considerably in neurons of supraoptic and paraventricular nuclei during endotoxin induced fever. The development of hypothermia was characterised by a decrease in metabolic activity of neurons of supraoptic, paraventricular and medium preoptic nuclei. It is supposed that differently directed metabolic activity changes in neurons of anterior hypothalamic nuclei during hyperthermia are connected with the mechanisms of body temperature regulation (median preoptic nuclei) and neurosecretory processes (supraoptic and paraventricular nuclei). PMID- 11868264 TI - [Effect of ryodipine, fenigidin, verapamil, and manganese ions on the contractile strength and the myometrium potential duration during rhythmic stimulation and at rest]. AB - Action of the calcium channels blocking agents was studied using their strong but reversible concentrations, in different regimens. Riodypine and phenihydine suppressed the contraction strength and shortened the AP duration in the myometrium muscle stripes equally effectively both in resting and under stimulation (0.3 Hz). Verapamyl and manganese ions suppressed the contraction strength and shortened the AP duration more effectively under rhythmic stimulation than in resting. PMID- 11868265 TI - [Adaptive thermoregulatory reactions of the rabbit fetus at the end of the intrauterine development]. AB - The data of permanent simultaneous registration of electrocardiogram, rectal & brown adipose tissue temperatures of the rabbit foetus have been obtained in chronic experiments. The haemodynamic shift in intensity of maternal-placental and foeto-placental blood flow (by female trental injection) led to a decrease in the foetus rectal temperature (0.33 +/- 0.09 degree C in intact foetus and 0.58 +/- 0.27 degree C in retarded foetus, p < 0.05). Both foetuses reacted by rectal temperature decrease (0.65 +/- 0.28 & 0.68 +/- 0.31 degree C, respectively) during immobilisation (by foetus arduan injection). Thus in both series of experimental cooling of foetus, brown fat activation was similar in intact foetus (approximately 53%), but did not change in growth retardation foetus--as a result of its tissue functional immaturity, probably. PMID- 11868266 TI - [Effect of electrical stimulation of the trigeminal nucleus caudatus on the blood microvessel permeability in the eye in normal and capsaicin-treated rats]. AB - Electrical stimulation of the trigeminal caudate nucleus increased permeability of the eye blood microvessels in rats. Ten days after capsaicin pre-treatment (total dose 150 mg/kg, s/c for 2 days: 20, 30, 50, 50 mg/kg) the electrical stimulation did not evoke any changes of the microvascular permeability in the eye. The data suggest that the effect of the stimulation is mediated through capsaicin-sensitive neurones of the trigeminal ganglion. PMID- 11868267 TI - [Substance P as a factor enhancing resistance of the surfactant lung system to chronic immobilization stress]. AB - In chronic experiments on non-line white rats the influence of system (intraperitoneal) and central (into the lateral brain ventricle) administration of substance P on lung surfactant system was studied in stressed and non-stressed animals. A single injection of substance P limited pulmonary surfactant activity disorders in immobilisation stress. Stress-induced increase of phospholipids level in broncho-alveolar lavage fluid remained the same in intracerebroventricular administration and was partly reduced in intraperitoneal one. In intact rats, a single injection of peptide was accompanied by alveolar phospholipids accumulation. In rabbits, multiple intracerebroventricular injections of substance P enhanced the opposite effect. PMID- 11868268 TI - [Use of the "partition" test in the behavioral and pharmacological experiments]. AB - Application of partition test measuring the behavioural activity of animals near the transparent perforated partition (dividing the experimental cage onto two halves) as a reaction to the partner in the neighbouring compartment of common cage, is considered. Behavioural reaction as a response to other individual differs in dependence on psychological and physiological states, social experience, strains of the animals and kinds of partner. Partition test may be productive in experiments studying mechanisms of sociability, anxiety, aggressive and sexual motivations, olfaction, as well as for screening of psychotropic drugs. PMID- 11868269 TI - Hsp70 chaperone machines. PMID- 11868270 TI - Structure and function of the small heat shock protein/alpha-crystallin family of molecular chaperones. PMID- 11868271 TI - Structure, function, and mechanism of the Hsp90 molecular chaperone. PMID- 11868273 TI - Hsp70 proteins in protein translocation. PMID- 11868272 TI - The proteasome: a supramolecular assembly designed for controlled proteolysis. PMID- 11868274 TI - Prolyl isomerases. PMID- 11868275 TI - Catalysis of disulfide bond formation and isomerization in Escherichia coli. PMID- 11868276 TI - N-glycan processing and glycoprotein folding. PMID- 11868277 TI - Functional genomic approaches to understanding molecular chaperones and stress responses. PMID- 11868278 TI - The yeast prion [PSI+]: molecular insights and functional consequences. PMID- 11868279 TI - Clp ATPases and their role in protein unfolding and degradation. AB - Although much has been learned about the structure and function of Clp chaperones and their role in proteolysis, the mechanism of protein unfolding catalyzed by Clp ATPases and the mechanism of translocation of the unfolded proteins from Clp ATPases to partner proteases remain unsolved puzzles. However, models in which mechanical force is used to destabilize the structure of the substrate in a processive and directional manner are probable. It also seems likely that when ClpA ATPases are associated with proteases, unfolding is coupled to extrusion of the unfolded protein into the proteolytic cavity. In summary, it is anticipated that the large family of Clp ATPases will accomplish their many important cellular functions by similar mechanisms and what has been learned by studying the prokaryotic members reviewed here will shed a great deal of light on all members of the family. PMID- 11868280 TI - Allostery and protein substrate conformational change during GroEL/GroES-mediated protein folding. PMID- 11868282 TI - Is the fast food industry becoming the next tobacco industry? PMID- 11868281 TI - Type II chaperonins, prefoldin, and the tubulin-specific chaperones. PMID- 11868283 TI - From paleo-surgeons to cyber-surgeons. PMID- 11868284 TI - Impact of gastric banding on eating behavior and weight. AB - BACKGROUND: Laparoscopic adjustable gastric banding (LAGB) is now the most frequently performed gastric restriction procedure. While short- and long-term outcome have been described extensively, the relationship between eating behavior and weight reduction is still unclear. METHODS: The present study examined the eating behavior of 66 selected morbidly obese subjects before and after LAGB by means of the Eating Inventory (EI), the Binge Scale Questionnaire (BSQ) and additional items. Assessments were conducted 6 months before surgery and at 3, 9, and 12 months after surgery. RESULTS: Significant reductions of weight and BMI were observed until 9 months after surgery, with a loss of 34.5% of excess weight. Later, a slight weight regain was noted. At baseline, Cognitive Restraint (CR), Hunger (H) and Flexible Control (FC) were within the norm, whereas Disinhibition (D) and Rigid Control (RC) were significantly elevated. None of the scales were related to BMI. At follow-up, significant changes were observed between presurgery and 3 months follow-up (T2), with increased CR, FC, and RC, and decreased D, H, and Binge Eating. These changes mostly remained stable. The largest changes were observed 3 months post-surgery in Flexible Control (FC), followed by D, CR, H, and RC. At follow-up, again no correlation was found between eating behavior and the total difference of BMI. CONCLUSION: LAGB results in significant reductions of weight, disinhibition and hunger during the first 9 months postoperatively. The change in eating behavior after surgery is independent of BMI and characterized mostly by elevated flexible control. Results indicate also changes of other behaviors that contribute to weight loss but are difficult to control. PMID- 11868285 TI - Eating patterns in morbidly obese patients before and after a gastric restrictive operation. AB - BACKGROUND: The present study was set up to analyze the relationships between eating patterns in morbidly obese patients who had undergone an adjustable silicone gastric banding (ASGB) followed for at least 2 years and morbidly obese patients without a gastric restrictive procedure. METHODS: Eating pattern was monitored by using the Dutch Eating Behavior Questionnaire in 99 morbidly obese patients (BMI > or = 35 kg/m2) preoperatively and in 31 patients who had undergone a stomach reduction by the Lap-Band followed at least 2 years. Both groups were compared to the Dutch normative scores. RESULTS: In the preoperative group, the scores on emotional eating and external eating were significantly higher than the Dutch normative scores. The scores on restrained eating were preoperatively equal to the Dutch normative scores. Although not significant, the scores in the postoperative group on external eating were lower than the Dutch normative scores and equal on emotional eating. The variable restrained eating postoperatively was significantly higher compared with the preoperative group. On emotional and external eating, the scores postoperatively were significantly lower compared with the preoperative group. CONCLUSIONS: According to the results, surgical treatment using an ASGB or another gastric restrictive operation could be the right solution in patients with an emotional and external eating behavior. Placement of the ASGB has a negative effect on restrained eating behavior. PMID- 11868286 TI - Impact of orlistat therapy on weight reduction in morbidly obese patients after implantation of the Swedish adjustable gastric band. AB - BACKGROUND: Adjustable gastric banding (AGB) is frequently performed to treat morbid obesity. One problem which can occasionally develop after a restrictive procedure is consumption of a high calorie liquid diet, which may prohibit further weight loss. Orlistat, a newly developed intestinal lipase inhibitor, prevents absorption of about one-third of ingested fat. It is unknown whether patients no longer losing weight after AGB, despite further band restriction, may lose weight with addition of orlistat. METHODS: 38 patients were selected who had stopped losing weight 3 months before the initiation of the study, 18 +/- 6 months (mean +/- SEM) after laparoscopic AGB. Subjects were divided into 2 groups, matched for age, sex, filling volume of the band and body mass index (BMI) both at the time of surgery and start of the study (18 +/- 6 months after AGB). RESULTS: Patients in group A received dietary counseling and orlistat 120 mg TID for 8 months, while patients in group B received only dietary counseling. During the following 8 months of study, subjects in group A lost 8 +/- 3 kg in weight, whereas subjects in group B lost 3 +/- 2 kg (p < 0.01, months 18 vs 26 of study; p < 0.03, group A vs B). In 15 patients from group A the study was further extended 9 months, but interestingly, weight remained stable independent of whether orlistat was continued (n = 8) or stopped (n = 7). 4 subjects were excluded from the extension study because of additional malabsorptive bypass surgery. Subjects taking orlistat encountered only minor GI side-effects. CONCLUSION: Orlistat appears to be useful when added in patients after AGB who are no longer losing weight, perhaps due to a high-calorie liquid diet rich in fat. PMID- 11868287 TI - Gastric cancer occurring after vertical banded gastroplasty. AB - A case of gastric cancer after vertical banded gastroplasty (VBG) is presented. A 44-year-old man presented with vomiting and weight loss 6 years after VBG. Endoscopy revealed a poorly differentiated gastric adenocarcinoma. The patient underwent a Whipple pancreaticoduodenectomy and received chemotherapy. He expired 6 months later. From our case and review of the literature, development of gastric cancer after VBG is very rare. The authors suggest that patients undergoing VBG be monitored by endoscopy after the operation. PMID- 11868288 TI - Preperitoneal herniation into a laparoscopic port site without a fascial defect. AB - BACKGROUND: Port site herniation is an uncommon event that usually occurs as a result of incomplete fascial closure. This allows the omentum or viscera to herniate through the incompletely closed defect. However, in laparoscopic surgery for morbid obesity, the omentum and viscera can herniate through the thick preperitoneal space even with a complete closure of the fascia. CASE REPORT: A 19 year-old female with BMI 55 underwent uneventful long limb laparoscopic Roux-en-Y gastric bypass. On postoperative day 1 the patient had limited pain, was ambulating well, and was tolerating slps of liquids. A limited upper GI series performed on postoperative day 2 revealed no leak or obstruction. Several hours later the patient developed abdominal pain associated with nausea, which progressed to vomiting. CT of the abdomen suggested a port site herniation into the left subcostal port. The cause of the obstruction appeared to be herniation through the left subcostal port site. At laparotomy, a segment of bowel just distal to the anastomosis was found herniated through the port site. The Richter's hernia was reduced. Careful inspection of the fascia revealed a complete fascial closure, with the strangulated portion of the bowel incarcerated in the preperitoneal space. Following repair of the preperitoneal defect, her subsequent recovery was unremarkable. CONCLUSION: Laparoscopic surgery for morbid obesity presents the possibility for preperitoneal herniation. Closure, using a fascial closure device, under laparoscopic control, may offer a solution by closing both the fascia and peritoneum all at once. PMID- 11868289 TI - Left subcostal minilaparotomy in silastic ring vertical gastroplasty and transected Roux-en-Y gastric bypass. PMID- 11868290 TI - A study of discrimination within the medical community as viewed by obese patients. AB - BACKGROUND: Although prejudice may not be verbal in nature, the lack of response from professional and non-professional medical personnel regarding the obese patients' needs (medical equipment, comfortable surroundings, properly fitting attire, etc.), leads one to assume that obese patients continue to be a target of unfavorable opinion. METHODS: 200 patients from four busy east coast bariatric practices received detailed surveys requesting information regarding experiences incurred during the entire peri-operative period. The responses were blinded as to patient or site of surgery. Patients were questioned about attitudes of primary care and consulting physicians and staff, all hospital and outpatient departments, and patient care areas within the hospital. The appropriateness of equipment was also questioned. RESULTS: There were 40 respondents aged 44.7 years (range 21-61 years); 34 were women. Average preoperative weight was 145 kg (range 101.8-231.8 kg). Average weight loss at time of survey was 42.8 kg; average time after surgery was 9.5 months. Responses concerning physician attitudes were similar through all patient groups. There was no difference in degree of discrimination felt by the lightest or the heaviest of the morbidly obese. 7 patients (17%) changed primary care physicians due to a perceived indifference, lack of concern, or negative attitude toward bariatric surgery. The procedure oriented physicians (orthopedic surgeons and gastroenterologists) were most supportive, while social workers and psychologists scored lowest. Most hospital departments treated patients well, except those with equipment that may be affected by increased patient weight (e.g. treadmills and x-ray tables). CONCLUSION: Patients undergoing bariatric surgery continue to feel misunderstood and mistreated by medical and non-medical personnel involved in the treatment of their obesity. Like other forms of prejudice, this most likely is due to a lack of understanding of the disease of morbid obesity, the root causes and the medical consequences if untreated. Despite laws designed to prevent discrimination based on appearance, unfavorable attitudes and practices persist. A plan for continued education of the medical and non-medical communities is essential to breakdown the barriers in place due to ignorance and indifference. Patient support groups continue to play an important role in the ongoing battle to correct the negative effect of these attitudes on the morbidly obese patient. PMID- 11868291 TI - A comparison of two different prophylactic dose regimens of low molecular weight heparin in bariatric surgery. AB - BACKGROUND: Deep venous thrombosis (DVT) is a significant risk in patients undergoing surgery for morbid obesity and may be associated with significant morbidity and mortality. In a consecutive group of patients in one bariatric surgery practice, the initial group of patients who received prophylaxis for DVT was given enoxaparin 30 mg q12h while the later group was given enoxaparin 40 mg q12h. METHODS: 481 patients who underwent primary and revisional bariatric surgery over 38 months (October 1997-December 2000) were evaluated. All patients received a multi-modality DVT prophylaxis protocol that included: early ambulation, graduated compression stockings, intermittent pneumatic compression, and enoxaparin (LMWH) in two dosage groups. The first 92 patients (19%) in the series (Group I) received LMWH 30 mg q12h while the subsequent 389 patients (81%) (Group II) received LMWH 40 mg q12h. RESULTS: Group I patients were not different from Group II patients in body mass index (BMI) (51.7 vs 50.3 kg/m2), age (43.7 vs 44.3 yrs), sex (men 20.2% vs 15.8%) or history of previous DVT (3.2% vs 3.9%). Group I patients did have significantly longer procedure times (213 vs 175 min, p < 0.05) and hospital stays (5.67 d vs 3.81 d, p < 0.05) than Group II. There were a total of 7 (1.4%) postoperative DVT complications. 5 DVT complications occurred in Group I (5.4%) compared with 2 DVT complications in Group II (0.6%) (p < 0.01 by Fisher Exact Test two-tailed). One patient in each group required treatment for hemorrhage. CONCLUSION: A multi-modality prophylaxis treatment protocol in patients undergoing bariatric surgery is feasible and achieves a low incidence of postoperative DVT complications. The use of a higher dose of enoxaparin, 40 mg q12h, may reduce the incidence of DVT complications in patients following bariatric surgery without an increase in bleeding complications. PMID- 11868292 TI - A genetic "obesity risk index" for patients with morbid obesity. AB - BACKGROUND: The influence of genetics on obesity is well established. Adoption studies and twin studies suggest that about 80% of the obesity risk is genetic. We designed a tool to predict outcomes of treatments in patients with sporadic or familial obesity. METHODS: Two factors best correlate with multifactorial genetic risk: 1) familial history and 2) age of onset. 147 morbidly obese adults self- or physician-referred for possible surgery for morbid obesity (age 17-66 y, BMI 35 82) were studied. Six elements were selected to measure the genetic influence on patients' weight: 3 personal weight milestones (weight at age 10, 20 and 30), and 3 family history factors (parents' weight, siblings' weight and second degree relatives' weight. These 6 elements of personal and family history information were collected prospectively on 35 obese patients and a feasible scoring system devised, with 0 points signifying no genetic component and 100 points suggesting the maximal possible genetic risk for obesity. Prospective data were then collected on 147 consecutive patients seen in consultation for possible bariatric surgery, to provide this "obesity risk index" (ORI). RESULTS: The final scoring system for the ORI assigned 50 possible points for personal weight milestones and 50 possible points for family history factors. At age 10, patients receive 10 or 20 points for being 2 or 3 SD above the mean BMI for age, respectively. At age 20, 10 or 20 points are received for BMI > 30 or 40, respectively. At age 30, 5 or 10 points are received for BMI > 35 or 50, respectively. 0 to 28 points are awarded for parental obesity, with 7 or 14 points for each parent with BMI > 30 or 40, respectively. The mean BMI of all siblings was calculated, with 6 or 12 points received for mean BMI greater than 30 or 40, respectively. Two points are awarded for each second degree relative with BMI > 35, to a maximum of 10 points. The mean (+/- SEM) score for our first 114 patients was 32 +/- 2 (range 0 to 87). The median score was 28.13% of patients had scores < 10; conversely, 13% scored points on all 6 elements. CONCLUSION: An ORI has been devised to quantify the genetic contribution to an individual's weight. Using this scoring system, we found that about 85% of patients who are candidates for bariatric surgery have elements in their history to suggest a genetic risk for morbid obesity. About 15% have extremely strong genetic ORIs. PMID- 11868293 TI - The Surgeon-General's call to action to prevent an increase in overweight and obesity. Released Dec. 13, 2001. PMID- 11868294 TI - Peptic ulcer/stricture after gastric bypass: a comparison of technique and acid suppression variables. AB - BACKGROUND: Mason's original animal experiments on the gastric bypass (GBP) showed little acid production in the gastric pouch, a finding confirmed in humans. Despite this, GBP in humans is associated with an incidence of ulcer/stricture (U/S) at the gastrojejunostomy of 3 to 20%, with both acid secretion and staple-line dehiscence considered important risk factors or etiologies. Our series of GBP patients was reviewed to determine what technical or management factors, if any, were associated with U/S. METHODS: All patients undergoing first time GBP at Dartmouth-Hitchcock Medical Center by one surgeon from June 1991 until June 2000 were reviewed. The incidence of U/S as confirmed on upper endoscopy was determined by retrospective chart review. The technique of surgery, frequency of acid suppressive therapy at discharge, postoperative day of U/S diagnosis by endoscopy, length of follow-up with a member of the multidisciplinary bariatric team, and incidence of staple-line dehiscence were tabulated. RESULTS: 158 patients (72% female, mean BMI 53, mean age 42) underwent GBP. Two gastric stapling methods were used to create the gastric pouch: 4-rows (136 patients) and 8-rows (22 patients). No other technical feature was adjusted in the series. The two patient groups were similar in gender, age, and BMI. Acid suppressive therapy at the time of discharge was similar in each group with U/S (4-rows 64% and 8-rows 50%, p = 0.5). U/S developed in 12 (55%) of the 8-row group and in 14 (10%) of the 4-row group (p < 0.001). U/S typically occurred within the first 2 months postoperatively (mean 48 days, SD 40). No patients in our series developed a staple-line dehiscence. CONCLUSION: U/S occur in the first few months following GBP. Twice the number of gastric staple-lines is associated with over five times the incidence of U/S, whereas post-discharge acid suppressive therapy is not predictive of U/S. Thus, a technique performed to decrease the risk of staple-line breakdown was associated with a much higher incidence of U/S. Staple-line dehiscence is not the etiology of this condition. Therefore, U/S after GBP does not appear to be explained by acid injury. We speculate that local, tissue injury related factors may be more responsible, a speculation that invokes a novel pathophysiologic mechanism for U/S formation following gastrojejunostomy. PMID- 11868295 TI - The utility of contrast studies and drains in the management of patients after Roux-en-Y gastric bypass. AB - BACKGROUND: The role of routine post-operative contrast examination (UGI) and drainage of the gastrojejunostomy after Roux-en-Y gastric bypass (RYGBP) is controversial. The authors determined if early routine post-operative UGI detects occult anastomotic leaks, thereby altering treatment and withholding early feeding. METHODS: Prospective data on 100 consecutive patients who underwent RYGBP from September 1998 to September 2000 was reviewed. Closed suction drains were routinely used. Within 36 hr post-operatively, all patients underwent UGI to evaluate the gastrojejunostomy. Patients were given liquids if the UGI showed no leak, and drains were removed 24 hr later. A blinded radiologist reviewed all the UGI. RESULTS: 87 women and 13 men underwent 75 open and 25 laparoscopic RYGBP. BMI was 52.0 kg/m2. 3 patients whose UGI showed a leak were treated non operatively with antibiotics, maintenance of drains, nasogastric tube and NPO. 2 of those patients developed purulent drainage within 24 hr after the UGI. None of the three patients required reoperation. 4 UGI were not available for the blinded reviewer who graded the remaining as satisfactory (94) and unsatisfactory (2). This reviewer disputed a leak in 1 of 3 previously reported leaks and reported a leak in a previously negative study. The latter patient subsequently required surgery for an uncontrolled leak. CONCLUSIONS: UGI can be used to withhold early oral intake in patients with radiographic leaks that would otherwise progress to clinically significant leaks. Surgical drains facilitate the non-operative management of such anastomotic leaks. Planned early UGI and surgical drains minimize the morbidity of anastomotic leaks after bariatric surgery. PMID- 11868297 TI - Liver pathology in morbidly obese patients undergoing Roux-en-Y gastric bypass surgery. AB - BACKGROUND: Non-alcoholic fatty liver disease is common. However, little is known about liver disease in the morbidly obese. METHODS: 75 subjects (78% female, mean BMI 57 [40-108]) who had intra-operative liver biopsies at the time of Roux-en-Y gastric bypass surgery were studied. RESULTS: 84% of subjects had steatosis while only about 20% had moderate to severe inflammation and fibrosis. 8% had bridging fibrosis or cirrhosis. The presence of fibrosis correlated strongly with the presence of inflammation (p < 0.001) and steatosis (p = 0.0011), but weakly with ALT (p = 0.02) and not with AST (p = 0.12) or with BMI (p = 0.34). Steatosis correlated with AST (p = 0.04) and ALT (p = 0.055), but not with BMI. CONCLUSION: Liver disease is not rare in the morbidly obese. The exact causes and mechanisms that lead from the very common isolated steatosis to inflammation and fibrosis remain unclear. Intra-operative liver biopsies during bariatric surgery may be helpful to screen for the presence of steatohepatitis and fibrosis. PMID- 11868298 TI - Pain, privilege and responsibility. PMID- 11868296 TI - Virtual gastroduodenoscopy: a new look at the bypassed stomach and duodenum after laparoscopic Roux-en-Y gastric bypass for morbid obesity. AB - BACKGROUND: After open or laparoscopic Roux-en-Y gastric bypass (RYGBP) for morbid obesity, the bypassed stomach and duodenum are not readily available for radiological and endoscopic evaluation. Furthermore, little is known about the long-term physiologic and histologic changes that occur in the bypassed GI segments following these procedures. Many alternative radiological and endoscopic techniques have been described to access the distal gastric pouch and the duodenum after RYGBP. Apart from percutaneous gastrografin studies, all these techniques require the insertion of a gastrostomy tube in the distal stomach. METHODS: A new diagnostic method to access the bypassed segments by virtual CT gastroscopy (VG) was used in 5 morbidly obese patients who underwent laparoscopic RYGBP (LRYGBP). RESULTS: All patients tolerated the procedure well, which appears safe and suitable for an outpatient setting. The virtual images offered an excellent intraluminal view of the stomach and duodenum. CONCLUSIONS: VG holds promise as the method of choice in the follow-up of LRYGB patients, having the potential to detect inflammatory changes and cancer in the excluded segments early. PMID- 11868299 TI - Gastric bypass in patients with BMI < 40 but > 32 without life-threatening co morbidities: preliminary report. AB - BACKGROUND: Surgical intervention is currently indicated for patients with BMI > 40 or > 35 with life-threatening comorbidities. Patients with BMI 32-40 without these comorbidities not only have the increased propensity to develop them but also suffer from similar psychosocioeconomic consequences. These patients may not respond to non-surgical treatment of obesity any better than those with BMI > 40. The question has been raised whether to offer them surgical intervention. METHODS: A study was carried out to determine outcome of surgery on patients with BMI > 32 but < 40 without life-threatening comorbidities but with either psychological, economic or social impairments affecting their quality of life. The approval of our Hospital Internal Review Board was obtained. In addition to routine evaluation for surgical intervention, these patients were required to have the approval of their primary care physician, be seen pre-operatively by a psychiatrist, and have a member of the family or a very close friend present at the time of discussion of operative risks and follow-up requirements. Patients committed to at least a 5-year follow-up. They were to be self-paying patients. The transected silastic ring vertical gastric bypass with a temporary gastrostomy was used. RESULTS: 50 patients, 49 women and one man, were entered into the study between May 1, 1999 and April 30, 2000. 50% were self-pay, and the other 50% were able to obtain coverage through their insurance companies. There were few peri operative complications and no deaths. The late complications include incisional hernias, dumping and transient alopecia. Hospital stay averaged 3.7 days. Follow up has been 18-27 months. Weight loss has been excellent. CONCLUSION: Preliminary results of surgical intervention extended to patients with BMI 32-40 without life threatening comorbidities but with psychosocioeconomic ramifications are very promising. Long term follow-up and comparison with other bariatric patients are planned. PMID- 11868300 TI - Radiology of patients with vertical banded gastroplasty. AB - BACKGROUND: The authors evaluated the usefulness of routine traditional radiology in the management of patients submitted to VBG. METHODS: Radiological findings in 65 subjects who had undergone VBG were evaluated. Post-surgical clinical and radiological examinations were performed 3 days, 1, 4 and 12 months after surgery. Upper GI symptoms, gastroesophageal imaging and 80% solid meal pouch emptying time were recorded. RESULTS: The routine postoperative study, in the absence of clinical symptoms, showed no unsuspected complication in any patient, both in the early and in the late postoperative period. There was no significant correlation at 4 and 12 months between emptying time and duration of satiation, emptying time and percent excess weight loss (%EWL), and duration of satiation and %EWL. CONCLUSIONS: Traditional radiological studies can be safely omitted from both the immediate postoperative period and from the long-term follow-up in asymptomatic VBG patients. The studies were not helpful in understanding functional changes leading to weight loss after VBG. PMID- 11868301 TI - Laparoscopic vertical banded gastroplasty: early results with the JOVO procedure. AB - BACKGROUND: Vertical banded gastroplasty (VBG) is an established treatment for morbid obesity for selected patients. This study seeks to assess the effectiveness of a laparoscopic version, the JOVO procedure, of the VBG. METHODS: An Independent surgeon interviewed all patients, who had had the JOVO procedure at one institution, evaluating weight loss, comorbid conditions, satisfaction and quality of life using the 36-item short-form health survey (SF-36). RESULTS: 14 JOVO procedures were done by two surgeons. Mean body mass index was 44 kg/m2, mean age 30 years and each patient had at least one comorbid condition. Mean operative time was 165 minutes. There was 1 complication, a suspected gastric leak requiring reoperation. Excluding this patient, mean hospital stay was less than 48 hours. Mean weight loss 4 weeks after surgery was 9 kg or 18% of excess weight. Mean excess weight loss of the 5 patients available for 1-year follow-up was 42% at 6 and 85% at 12 months; 1 did not continue to lose weight. All but 1 preoperative comorbid condition resolved or improved. 13 of the 14 patients were fully satisfied. SF-36 scores were high in the 13 that lost weight, especially in physical and social functioning areas. CONCLUSION: The JOVO procedure is safe and reproduces laparoscopically the early weight loss of open VBG with much shorter hospital stay and low complication and failure rates. Longer follow-up and larger numbers are needed. PMID- 11868302 TI - Body image: appearance orientation and evaluation in the severely obese. Changes with weight loss. AB - BACKGROUND: The severely obese experience discrimination and embarrassment regarding their appearance, causing psychosocial distress. We assessed the importance of appearance, presentation and self-evaluation of appearance before and after weight loss, in severely obese subjects (BMI > 35 kg/m2). METHODS: Appearance orientation (AO) and appearance evaluation (AE) sections of the Multi dimensional Body Self Relations Questionnaire were completed by preoperative patients and those attending an annual follow-up after Lap-Band gastric restrictive surgery over a 2-year period. AO is a measure of the importance one places in appearance. AE is a self-evaluation of one's appearance. RESULTS: Before surgery 322 consecutive patients (48 men and 274 women) completed the survey. AO was similar to that of community norms (mean scores 3.76 +/- SD 0.6 vs 3.84 +/- 0.6), with the exception of the super obese (BMI > 50 kg/m2) who placed significantly less importance on their appearance (3.34, +/- 0.8, p = 0.001). There was no change in AO at 1-4 years after surgery, with the exception of the super obese whose mean values rose to normal by 1 year. AE was very low before surgery compared with community normal values (1.6 +/- 0.6 vs 3.4 +/- 0.8, p < 0.001). Major improvement in mean AE was seen at 1 year after surgery (n = 209, 2.6 +/- 0.8, p < 0.001) and the improvement was maintained out to 4 years. The change in AE from pre-surgery to 1 year (n = 122 paired) correlated positively with the percentage of excess weight lost (r = 0.32, p < 0.01) and positively with measures of quality of life and psychological disturbance. CONCLUSION: Major improvements in appearance evaluation occur with weight loss after surgery and this is associated with psychological benefit. PMID- 11868303 TI - Esophageal motility and reflux symptoms before and after bariatric surgery. AB - BACKGROUND: Surgical treatment is the most effective method for weight reduction in morbid obesity. The most common operations are gastric banding and gastric bypass. The effect of these interventions on esophageal function and gastroesophageal reflux symptoms has not been adequately investigated. METHODS: Patients undergoing obesity surgery were prospectively included in an observational study. Before surgery, each of the 53 patients underwent pulmonary function tests, esophageal manometry, and gastroscopy. Drug medication and esophageal symptoms were recorded. "Non-sweet eater" patients with good compliance underwent laparoscopic adjustable gastric banding (LAGB). In "sweet eating" or non-compliant patients, gastric bypass (GBP) was carried out. RESULTS: Between July 1997 and April 2000, 53 patients (9 males and 44 females) were consecutively operated on. 32 patients (median BMI 46.4 kg/m2 +/- 5.4 SD) received LAGB, and 21 patients (BMI 54.0 kg/m2 +/- 10.7) GBP. Median follow-up was 22 months, and only 3 patients were lost to yearly follow-up. Preoperatively, 6 LAGB patients had reflux symptoms, which postoperatively resolved in 3 of them, while the other 3 noted no change. Three patients who had no preoperative reflux symptoms developed them after LAGB. In the GBP group, no patient had esophageal dysmotility or incompetent esophageal sphincter function pre- or postoperatively. The incidence of postoperative esophageal symptoms was independent of operative technique (Wilcoxon U-Test: p = 0.75). CONCLUSION: The present results do not show any effect of gastric reduction surgery on postoperative esophageal function or gastroesophageal reflux symptoms. PMID- 11868304 TI - Modifications of metabolic and cardiovascular risk factors after weight loss induced by laparoscopic gastric banding. AB - BACKGROUND: The well-known inverse relation between life expectancy and BMI, particularly in morbid obesity, is presumably in large part due to multiple cardiovascular and metabolic comorbidities. Severely obese patients treated with laparoscopic adjustable silicone gastric banding (LASGB) were evaluated for such risk factors before and 1 year after LASGB. METHODS: 130 individuals (age 20-66, BMI 34-59 kg/m2) who underwent LASGB between 1996 and 2000 were studied; 50 of them were available for reevaluation 12 months after surgery. The presence and severity of diabetes (DM), hypertension (HTN), hypercholesterolemia (HC) and hypertriglyceridemia (HT) were assessed before and after surgery. In 18 of them erythrocyte sedimentation rate (ESR) were also measured. RESULTS: Comorbidities were highly prevalent at the initial evaluation: DM 10%, HTN 32%, HC 37%, HT 27%. In the subgroup reevaluated after surgery, prevalence of DM decreased from 15% to 6%, HTN from 37% to 25%, HC from 36% to 25%, and HT from 29% to 13%, with an average BMI loss from 44.1 to 35.9. ESR decreased from a preoperative value of 36.7 +/- 22.6 mm/hr to 18.3 +/- 11.9 mm/hr at 1 year (p < 0.001). CONCLUSION: Morbidly obese subjects respond to LASGB with an impressive reduction of comorbidities which is sustained for at least 1 year, well after the initial acute negative energy balance and when weight tends to stabilize. The high prevalence of elevated ESR, which has been considered a strong predictor of coronary mortality, confirms previous reports of its association with obesity, and the clear tendency to normalization with weight loss may represent a further element contributing to lower morbidity. PMID- 11868305 TI - Outcome predictors in morbidly obese recipients of an adjustable gastric band. AB - BACKGROUND: The authors investigated the outcome predictors in obese patients who underwent laparoscopic adjustable banding with the Lap-Band. METHODS: The 3-year excess weight loss (EWL) and rate of band-related complications (pouch dilatation and port leakage) were analyzed in 260 morbidly obese patients, according to several possible predictive characteristics. Success rate (EWL > 50%), failure rate (EWL < 20%) and weight regain rate (regain of > 10% EWL between 1 and 3 yrs) were considered. RESULTS: The Lap-Band produced a 43.0 +/- 22.3% EWL, corresponding to a BMI reduction from 46.6 +/- 7.0 to 36.8 +/- 6.6 kg/m2. Success rate was 35.7%, failure rate was 14.1% and weight regain rate was 20.7%. Pouch dilatation occurred in 32 patients (12.3%), band erosion in 2 (0.8%), port leakage in 74 (28.5%), and port twisting in 2 (0.8%). Major band-related surgery was requested in 11 patients (4.2%) and minor port-related surgery in 62 patients (23.9%). Significant success predictors were found to be age < 40 years and BMI < 50 kg/m2. Significant failure predictors were found to be male sex and non-sweet eating behavior. Significant weight regain predictors were found to be BMI < 50 kg/m2 and the occurrence of a port leakage. Port leakage was significantly more frequent in women and in patients with BMI < 50 kg/m2. The prevalence of pouch dilatation was threefold higher in women than in men. CONCLUSIONS: Lap-Band was associated with a good outcome and with a low rate of severe complications. The outcome was more influenced by physiological and technical reasons than by psychological or behavioural factors. PMID- 11868306 TI - Adjustable gastric banding in a public university hospital: prospective analysis of 400 patients. AB - BACKGROUND: Laparoscopic application of an adjustable gastric band (LAGB) is considered the least invasive surgical option for morbid obesity. It has the advantage of being potentially reversible and can improve quality of life. METHOD: Between April 1997 and January 2001, 400 patients underwent LAGB. There were 352 women and 48 men with mean age 40.2 years (16-66). Preoperative mean body weight was 119 kg (85-195) and mean body mass index (BMI) was 43.8 kg/m2 (35.1-65.8). RESULTS: Mean operative time was 116 minutes (30-380), and mean hospital stay was 4.55 days (3-42). There was no death. There were 12 conversions (3%). 40 complications required an abdominal reoperation (10%), for perforation (n = 2), gastric necrosis (n = 1), slippage (n = 31), incisional hernia (n = 2) and reconnection of the tube (n = 4). We noticed 7 pulmonary complications (2 ARDS, 5 atelectasis) and 30 minor problems related to the access port. At 2 years, mean BMI had fallen from 43.8 to 32.7 kg/m2 and mean excess weight loss (EWL) was 52.7% (12-94). CONCLUSION: LAGB is a very beneficial operation with an acceptable complication rate. EWL is 50% at 2 years if multidisciplinary follow up remains assiduous. Surveillance for late anterior stomach slippage within the band is essential. PMID- 11868307 TI - How to taper tramadol dose. PMID- 11868308 TI - Systemic complications of chronic kidney disease. Pinpointing clinical manifestations and best management. AB - The kidney plays a critical role in the maintenance of homeostasis. As kidney function diminishes, excretory, regulatory, and endocrine function is lost, and complications develop in essentially every organ system. Kidney failure is the last stage in the continuum of progressive CKD. Management of the complications associated with CKD mainly includes dietary counseling, adequate control of volume and blood pressure, and use of phosphate binders, calcitriol (Calcijex, Rocaltrol), and erythropoietin. Many of these complications can be prevented or attenuated with optimal CKD care, which involves early detection of progressive kidney disease, interventions to retard its progression, prevention of uremic complications, attenuation of comorbid conditions, adequate preparation for kidney replacement therapy, and timely initiation of dialysis (figure 2). Closer attention to CKD care is likely to be the key to improved outcomes among patients with kidney failure. PMID- 11868309 TI - Nocturnal leg cramps. PMID- 11868310 TI - Mad cow disease risk in the United States. Does perceived threat overshadow true likelihood of occurrence? PMID- 11868311 TI - What not to do after a whiplash injury. PMID- 11868312 TI - Progressive, painful hardening of the legs. Lipodermatosclerosis. PMID- 11868313 TI - Evaluating the comatose patient. Rapid neurologic assessment is key to appropriate management. AB - Coma is defined as a sleeplike state in which the patient is unresponsive to self and the environment. Coma should be distinguished from the persistent vegetative state and locked-in syndrome. It is important to obtain a carefully taken history from eyewitnesses and to perform a rapid neurologic examination focusing on pupillary responses, eye movements, and motor responses. Pupils reactive to light usually indicate metabolic or medical coma; cerebellar infarction or hemorrhage is a notable exception. A pupil unreactive to light often points to a structural brain lesion and the need for urgent neurosurgical consultation. The prognosis for coma depends on the cause. PMID- 11868314 TI - Sedation and analgesia in intensive care. Medications attenuate stress response in critical illness. AB - The stress response to critical illness can have many deleterious effects. Appropriate use of sedation and analgesia can attenuate the stress response, alleviate pain and anxiety, and improve compliance with care. Agitation responds best to anxiolytic drugs; pain is best relieved by analgesics. A combination of these drugs can act synergistically, because most analgesics provide some degree of sedation. In select cases, neuromuscular blocking agents are required, but they should not be used without concomitant sedation and analgesia. Use of agents needs to be tailored to the needs of individual patients; indications, anticipated length of need, and underlying organ system derangements are important considerations. PMID- 11868316 TI - Management of common stress fractures. When to apply conservative therapy, when to take an aggressive approach. AB - Stress fractures are an increasingly common injury in primary care patient populations. Early diagnosis and prompt conservative therapy are effective for most of these injuries. Special attention to clues from history taking and physical examination in an at-risk patient population can lead physicians to the correct diagnosis. Most stress fractures resolve with rest and progressive reintroduction of stressors, but certain injuries, such as stress fractures of the femoral neck and fifth metatarsal, have a high risk of complications and require aggressive therapy. PMID- 11868315 TI - Who's liable for breast cancer prevention? Your patient can sue--and win--if preventive options aren't made clear. AB - Strategies to reduce the likelihood of breast cancer in high-risk patients present complex medical-legal issues. An accurate and thorough medical history, objective calculation of risk, thorough discussion of preventive options and side effects, meticulous documentation of physician-patient interactions, and meticulous follow-up are essential. These elements form the foundation of a sound approach to breast cancer prevention in high-risk patients and should reduce physician liability if cancer occurs. PMID- 11868317 TI - [The contribution of professor Antoni Pruszewicz into Polish and European audiology and phoniatry]. PMID- 11868318 TI - [Paclitaxel (Taxol) in chemotherapy of head and neck cancer--just another proposal or breakthrough?]. AB - The examples of so far application of taxol in head and neck cancer are not completely clear. This is the first representative of a totally new group of cytostatics known as taxones. Presently, paclitaxel (TAXOL) is the anti-cancer drug commonly used in ovary cancer patients, resistant to platinum analogues. Taxones have the different functioning than the other anti-cancer drugs. They are responsible for originating microtubules and their further stabilisation. Moreover, paclitaxel (TAXOL) is a chemical compound sensitizing tumor cells for ionizing radiation. The authors are trying to specify the role of taxones nowadays and for the future use. PMID- 11868319 TI - [Value of estradiol, progesterone and cortisol binding globulin (CGB) in patients with laryngeal cancer]. AB - The unique feature of larynx cancer epidemiology is great discrepance between men and women morbidity. This difference may be explained not only by exposition to environmental factors but also to endogenous one, i.e. sex hormone levels. In 67 patients operated for larynx cancer there were simultaneously estimated the value of estradiol (E2), progesterone (Pg) and cortisol binding globulin (CBG) in blood serum. The radioimmunological assay (RIA) with specific antibodies and antigens signed by J125 was used. Value of hormones and binding globulin were examined by Spectria sets by Orion Diagnostica. The high value of estradiol and CBG in blood serum was observed. In meaningful number of patients we noted normal value of progesterone. PMID- 11868320 TI - [Obstructive sleep apnea syndrome--the most common sleep related breathing disorder]. AB - Author has presented clinical classification of sleep related breathing disorders and discussed pathophysiology, clinical picture and diagnostic evaluation in Obstructive Sleep Apnea Syndrome--the most common syndrome among this group of disorders. PMID- 11868321 TI - [Therapeutic efficacy assessment of cefuroxime axetil in short 4 day course of empirical antibiotic therapy in patients with bacterial infections of upper respiratory tract and otitis media]. AB - There were examined 261 patients aged 18-62 ambulatory treated and divided into five groups: I--65 patients with acute maxillary sinusitis, II--43 patients with exacerbation of chronic maxillary sinusitis, III--40 patients with angina, IV--61 patients with acute otitis media and V--52 patients with exacerbation of chronic otitis media. Cefuroxime axetil have applied 2 times a day in 500 mg oral dose by 4 days. The therapeutic efficacy of cefuroxime axetil was assessed on the basis: fever chart and self assessment chart of showed symptoms which were assessed from 0 to 4.4 days course antibiotic therapy showed that the total receding of illness symptoms stated in 90.8% patients with acute maxillary sinusitis, in 69.8% patients with exacerbation of chronic maxillary sinusitis, in 62.5% patients with angina, in 91.8% patients with acute otitis media and receding of acute symptoms of exacerbation with chronic otitis media--a ear pain in 88.5% patients and a ear leakage in 44.2% patients. The obtained results stated that the 4 days course of cefuroxime axetil can be sufficient and efficacy in treatment of acute bacterial infections of maxillary sinuses and ear media. PMID- 11868322 TI - [Free forearm skin flap with microvascular anastomosis for oral cavity reconstruction]. AB - Free forearm skin-fascia flap was used in 8 patients for intraoral reconstruction. Tissue defect was located in anterior part of the oral cavity in three patients and in lateral part in 5 patients. In the first three patients microvascular anastomosis was performed first and the flap was sutured in the oral cavity later on. In one of these patients the whole flap and in two patients about half of the flap necrotised. In the next 5 patients oral defect was closed first and then microanastomosis was done with good result in all the patients. We advise the last type of reconstruction. Free forearm skin-fascia flap provides good anatomical and functional result of reconstruction in the oral cavity. PMID- 11868323 TI - [Minor congenital ear malformations: surgical management and results]. AB - The study present the results of surgical treatment of minor congenital anomalies of the middle ear in children. The ossiculoplastic procedures and the different types of stapedectomy carried out in the Department of Otolaryngology of Institute of the "Polish Mother's Hospital" in Lodz on 18 patients (11 girls and 7 boys) aged 5-16 years, with minor congenital anomalies of the middle ear. One month after the surgical procedure a lasting hearing improvement averaging 21.69 +/- 10.00 dB and 6 month after operation hearing improvement 21.91 +/- 9.75 dB was gained. The results of surgical procedures in minor congenital anomalies of the middle ear are not so good as in otosclerosis. PMID- 11868324 TI - [Objective evaluation of the influence of the hyoid bone resection on the disorders in the act of swallowing]. AB - Laryngographic, manometric and videorentgenocinematographic examinations of swallowing were conducted on 49 patients after partial laryngectomy and on 35 subjects being a control group. The results of examinations indicate that the mobility of remaining after the surgery parts of the larynx have the great influence on the efficient swallowing. The larynx mobility is closely related to the remaining of the hyoid bone. Resection of the hyoid bone or its part is the factor that causes intensified difficulty during swallowing and increase in the frequency of the occurrence of aspiration. The study of effectiveness of glottis or neoglottis performed with the use of laryngograph showed no relation between glottis/neoglottis occlusion effectiveness index and intensified swallowing disorders. This shows the lack of compensating option in swallowing disorders with the help of an mechanism based on an efficient glottis or neoglottis occlusion alone. PMID- 11868325 TI - Stenosis of the larynx and trachea: diagnostics and treatment. AB - 36 cases (26 males and 10 females aged 19-69, average age 40.3) of larynx and trachea stenosis (23/36 i.e. 64%) and occlusion (13/36 i.e. 36%) were observed between 1990-1999. The cause of larynx and trachea lesion was the prolonged intubation in 17 (47%) cases, tracheostomy--in 13 (36%) (in 12 cases performed outside our centre), external trauma in 5 cases (14%) and burns of the upper airway in 1 patient (3%). For the diagnosis endoscopic, radiologic and spirometric examinations were performed. Length of the constricted or occluded section was estimated at 5-65 mm. Patients were treated applying three methods: endoscopy, splitting of the larynx and/or trachea and "end to end" anastomosis. In most cases surgical proceeding was multistaged. The best result was obtain after "end to end" anastomosis (all patients were deccanulated). PMID- 11868326 TI - [Stapedectomy in the treatment of otosclerosis in children]. AB - In the years 1994-2000 in the Laryngology Department Silesian Medical Academy in Zabrze 167 stapedectomy were performed in 143 patients with otosclerosis. In this group of patients 11 stapedectomies in 8 children aged from 9 to 17 years were performed. 5.6% of all the patients with otosclerosis treated in our Department were children. The purpose of this study was to analyze the audiologic results of stapedectomies in children. The postoperative evaluations were performed in the period from 6 months to 6 years after the surgical treatment. In the estimation of the hearing state we analyzed the change of the air-bone gap, an average air conduction hearing improvement, the rate of percentage hearing improvement and an average hearing loss by Bell Telephone Laboratories. In all the operated children we observed the decrease of the air conduction hearing level. Stapedectomy was successful (postoperative air bone gap within 10 dB of the preoperative bone conduction) in 63.6% of the patients. In 3 cases the air bone gap was lower than 20 dB (27.3%). In one case it was greater then 20 dB. The air bone gap closure was at the average of 5.75 dB level. The results were comparable with the other authors. Stapedectomy can be an effective procedure for correcting conducting hearing losses due to the otosclerosis in the pediatric patients. PMID- 11868327 TI - [Management of nasal trauma in children]. AB - In this study we investigated the optimal treatment of nasal fractures and nasal deformations in children. An algorithm for the management of nasal fractures by children is publicated. PMID- 11868328 TI - [Epidemiology of acoustic neuromas in Poland]. AB - In 1996 the Polish Skull Base Surgery Society was founded and assembled mainly otolaryngologists and neurosurgeons from several dozen clinics and hospital wards. Its main aim was to encourage co-operation between those two specialties. For this reason among others a common project entitled: "Epidemiology of acoustic neuromas in Poland" was launched in 1997. Information was collected from questionnaires in seven neurosurgery and in three laryngology centres in the years from 1997-1998. So far, no epidemiology of acoustic neuromas was established for the population of Poland. In this span of time 72 patients were operated. The agreement upon similar criteria, terminology and ways of evaluating the treatment results was quite a problem. Most cases were found in patients in their fifties (19 cases) and sixties (15 cases), mostly women. The average duration of the symptoms was 43 months. Large tumours predominated in the study group--T4 in the Samii (67%), and > 26 mm in the Tos (74%) classification. Most commonly, the patients were subjected to computer tomography in the preoperative period. All patients were treated surgically, with the retrosinusoid approach predominating. Only in one case, the translabyrinthal approach was used. Authors of this work are all participants of the project. Despite the fact that the number of performed operations is not too high, the fact of co-operative studies on the results by representatives of those two specialties can be considered a success. Hopefully, in the next years it will gradually be possible to widen the scope of the project and obtain more precise information. PMID- 11868329 TI - [Does the electrodes insertion depth influence speech rehabilitation progress in CI patients?]. AB - Many interdisciplinary teams from difference CI centers are studying speech rehabilitation process of implanted patients. Speech rehabilitation progress depend on many conditions like: duration of deafness, age at implantation, cause of deafness. Rehabilitation of postlingually deafened patients is very short. These subjects return very fast to the world of sounds. By the other side speech rehabilitation of prelingually deafened patients needs much more time (years). There are although well seen differences in language skills acquisition inside these groups. It confirmed, that speech rehabilitation after cochlear implantation could depends on others conditions as well. The aim of this study was electrodes insertion depth evaluation (cochleograms) and its influence on speech rehabilitation progress. PMID- 11868330 TI - [Histopathological verifications clinical indications in the inferior turbinoplasty]. AB - Surgical treatment in cases of disturbances of nasal patency caused changes in the nasal inferior turbinates are controversial. The authors performed the microscopic (light microscope) and morphometric examinations of mucous membrane of the nasal inferior turbinates obtained after inferior turbinoplasty in patients with vasomotor and perennial allergic rhinitis and compensation hypertrophy of nasal inferior turbinate accompanied of nasal deviation septum. Analysing of morphologic changes in mucous membrane of the nasal inferior turbinates verified of clinic indications to inferior turbinoplasty. PMID- 11868332 TI - [Effectiveness of surgical treatment of laryngeal cancer depends on regional lymph node metastasis]. PMID- 11868331 TI - [Comparison of DNA adducts between oral, pharyngeal and larynx cancer]. AB - The results concerning examination of DNA adducts in oral (23 patients), pharyngeal (23 patients) and larynx cancer (10 patients) subjects are presented. DNA adduct levels were compared in respect to anatomical structure (primary tumour location), number of cigarettes smoked, TNM stage, and age of patients. DNA was isolated from removed tissue (tumour and non-tumour surrounding tissue) using detergent/phenol extraction. 32P-postlabelling assay including nuclease P1 enhancement modification was applied. Aromatic DNA adducts were found in all studied tissues. Total DNA adduct levels (tumour and non-tumour tissues) was lowest in larynx cancer, higher in oral cancer and highest in pharyngeal cancer. There were no influence of age into formation of DNA adducts. The higher level of DNA adducts was found in tumour tissue of oral cancer in the group of smokers with metastasis into lymph nodes. PMID- 11868333 TI - Generalized analysis of experimental data for interrelated biological measurements. AB - Important biological mechanisms, such as signal transduction and gene expression, are mediated by numerous interacting multifunctional molecules, whose expression and activation are tightly regulated in space and time in response to stimuli. In order to describe the network of functional inter-relationships that govern such mechanisms, we use simple algorithms to interpret multiple variable measurements, identify the prominent participants, evaluate their interactions and obtain a 'functional fingerprint' of cell behaviour. Dynamic measurements of responses yield hierarchical information about causal relations in the underlying pathway. As a proof of principles we apply this approach to phosphorylation assays in protein gels, probing hormone and insulin signalling. PMID- 11868334 TI - Qualitative analysis of the all-cause black-white mortality crossover. AB - To date, despite decades of investigations and the relative abundance of mortality data, our understanding of the phenomenon of 'mortality crossover' remains inadequate. We propose a methodology for transforming mortality data from the 'age-domain' to the 'time-domain'. We then introduce a model of selection partially offset by mobility, to simulate the dynamics of vulnerability in a population cohort that is heterogeneous in health and death. Using our model of vulnerability simulating the dynamics of mortality in the time-domain, we compare the mortality experience of the Black and White populations of the United States, identify the significance of selection and mobility as potential factors producing the crossover phenomenon, and make diagnostic use of them. PMID- 11868335 TI - Oxygen balance for small organisms: an analytical model. AB - An analytical model is developed that describes oxygen transport and oxygen consumption for small biological structures without a circulatory system. Oxygen inside the organism is transported by diffusion alone. Oxygen transfer towards the organism is retarded by a thin static fluid film at the surface of the organism. The thickness of this film models the outward water conditions, which may range from completely stagnant water conditions to so-called well-stirred water conditions. Oxygen consumption is concentration-independent above a specified threshold concentration (regulator behaviour) and is proportional to the oxygen concentration below this threshold (conformer behaviour). The model takes into account shape and size of the organism and predicts the transition from (pure) regulator behaviour to (pure) conformer behaviour, as well as the mean oxygen consumption rate. Thereby the model facilitates a proper analysis of the physical constraints set on shape and size of organisms without an active internal oxygen transport mechanism. This analysis is carried out in some detail for six characteristic shapes (infinite sheet, cylinder and beam; finite cylinder, sphere and block). In a well-stirred external medium, a flattened shape appears to be the most favourable for oxygen supply, while a compact shape (cube) is more favourable if the external medium is nearly stagnant. The theoretical framework is applied to oxygen consumption data of eight teleost embryos. This reveals relative insensitivity to external flow conditions in some species (e.g., winter flounder, herring), while others appear to rely on external stirring for a proper oxygen supply (e.g., largemouth bass). Interestingly, largemouth bass is the only species in our analysis that exhibits 'fin-fanning'. PMID- 11868336 TI - HIV-1 infection and low steady state viral loads. AB - Highly active antiretroviral therapy (HAART) reduces the viral burden in human immunodeficiency virus type 1 (HIV-1) infected patients below the threshold of detectability. However, substantial evidence indicates that viral replication persists in these individuals. In this paper we examine the ability of several biologically motivated models of HIV-1 dynamics to explain sustained low viral loads. At or near drug efficacies that result in steady state viral loads below detectability, most models are extremely sensitive to small changes in drug efficacy. We argue that if these models reflect reality many patients should have cleared the virus, contrary to observation. We find that a model in which the infected cell death rate is dependent on the infected cell density does not suffer this shortcoming. The shortcoming is also overcome in two more conventional models that include small populations of cells in which the drug is less effective than in the main population, suggesting that difficulties with drug penetrance and maintenance of effective intracellular drug concentrations in all cells susceptible to HIV infection may underlie ongoing viral replication. PMID- 11868337 TI - Adjustment of the human respiratory system to increased upper airway resistance during sleep. AB - A cardiorespiratory model incorporating control of the human upper airway during sleep is described. Most previous models have not considered the possibility that the upper airway could be a limiting factor for gas exchange. Our model was developed to also predict certain pathophysiological phenomena in the cardiorespiratory system that characterize heavy snoring or sleep apnea. We started by adapting our collapsible upper airway model to include the impact of nasal passage and larynx, and extended the model with equations for gas exchange in the lungs. A feedback loop both to the respiratory pump and the upper airway dilator muscles was included. The model enabled successful breath-by-breath simulations of obstructive events of the upper airway. Although the model incorporates several physiologically relevant components of the system, the simulation results suggest that only few parameters suffice to predict the key adjustments that the cardiorespiratory system is known to make in patients with heavy snoring. PMID- 11868338 TI - Lipoprotein oxidation and its significance for atherosclerosis: a mathematical approach. AB - Atherosclerosis is a chronic disease which involves the build up of cholesterol and fatty deposits within the arterial wall. This results in the narrowing of the vessel lumen, which eventually restricts blood flow to vital organs such as the heart and lungs. These events may culminate in a heart attack or stroke, the commonest causes of death in the U.K. population. In this paper we study the early stages of atherosclerosis which include the build up of cholesterol within subendothelial cells to form what is known as a fatty streak, the earliest identifiable evidence of atherosclerosis. The deposition of cholesterol is believed to be a consequence of oxidation of circulating cholesterol-rich lipoproteins, in particular low density lipoproteins (LDLs). Via a mathematical model we investigate this process of oxidation within the context of an in vitro framework. We first recreate existing experimental results and then extend the model to investigate phenomenon not studied by current experimental protocols. We find that the model displays hysteresis which reveals some interesting insights into possible in vivo events. Mathematical analysis of this behaviour predicts that vitamin E supplementation is not as beneficial as high density lipoproteins (HDLs) and vitamin C. Furthermore, the scavenging of oxidants by HDL can provide an important first line of defence against LDL oxidation. PMID- 11868339 TI - Physiologically based pharmacokinetic models for the transport of trichloroethylene in adipose tissue. AB - In this paper we present three physiologically based pharmacokinetic (PBPK) models for the systemic transport of trichloroethylene (TCE), with a focus on the adipose, or fat tissue. TCE is a widespread environmental contaminant, and has been shown to produce toxic effects in both animals and humans. A key characteristic of TCE is its tendency to accumulate in fat tissue, which has a major impact on the overall systemic disposition of TCE. Here we use PBPK models to predict the dynamics of TCE in the various tissues and organs, including the adipose tissue. The first model utilizes the standard 'perfusion-limited' compartmental model for the fat tissue, while the second model uses a 'diffusion limited' model to describe the transport through the adipose tissue. Both of these ODE models are based on 'well-mixed' and rapid equilibrium assumptions, and do not take into account the specific and largely heterogeneous physiology of adipose tissue. The third model we discuss is a PBPK hybrid model with an axial dispersion type model for the adipose tissue. This PDE-based model is designed to capture key physiological heterogeneities of fat tissue, including widely varying fat cell sizes, lipid distribution, and blood flow properties. Model simulations demonstrate that this model may be well-suited to predict the experimental behavior of TCE in adipose tissue using parameter estimation techniques. PMID- 11868340 TI - [The effects of stressors at work and in family life on self-rated health and psychological well-being]. AB - OBJECTIVE: This study examined the effects of stressors at work and in family life on self-rated health and psychological well-being. METHODS: The subjects were civil servants of T City (pop. 210,000) aged 35 years and older. In February 1998, a questionnaire survey was conducted (response rate: 82.6%). The 1,364 responders comprised 971 men, including 143 firefighters, and 393 women. Participants, excluding firefighters, were categorized into two groups according to satisfaction with their job and their family life. In these two groups, the stressors at work and in the home was examined. Job demands-control-support score, type of work, job rank, family control and close friends were examined with the t-test, and sex, hobbies and age examined with the chi 2 test. This was followed by covariance structure analysis of self-rated health and psychological well-being. RESULTS: For the groups dissatisfied with work or family life, the score for jpb demands was higher, and the scores for job control, social support and close friends were lower. The scores for self-rated health and psychological well-being items were also significantly lower. Covariance structure analysis demonstrated that factors at work had stronger effects. Regarding individual factors, job control, satisfaction with their job or family life and close friends had the greatest influence. CONCLUSIONS: Stressors at work and in family life result in lower self-rated health and psychological health. In particular, close friends and factors at work exert strongest effects on personal health. It is thus important for the prevention of stress-related diseases and the development of positive healthy life to reduce the work stressors and develop close friendships. PMID- 11868342 TI - [Public health and international health let us deepen our concern on international health]. PMID- 11868341 TI - [Cost-benefit analysis of community based rehabilitation program using willingness to pay measured by the contingent valuation method]. AB - OBJECTIVE: There have been few studies on economic evaluation of publicly provided community health services in Japan. The purpose of the present investigation was to conduct a cost-benefit analysis of a community based rehabilitation program provided by municipalities, using the willingness to pay (WTP) measured by the contingent valuation method, a survey-based approach to measure the monetary valuation for the benefit. METHODS: The study was held in 18 wards of Yokohama city. The cost per capita of the program in each ward was calculated using the data from a statement of accounts and a report of activities of public health nurses, other officials, and volunteers, in the 1999 fiscal year. The WTP for the program was measured by a survey of 631 people who participated in the programs held in October and November, 2000. The mean, median, and 95% CI of WTP were calculated as measures of the benefit. RESULTS: The response rate of the survey was 73.4%. The mean and median WTP were yen 441 and yen 300 respectively, and the 95% CI ranged from yen 800 to yen 1,682. The WTP was negatively related to the duration of functional disability, and positively related to income. However, the WTP was not significantly related to self-perceived effectiveness of the program and the SF-36. The costs per capita without the cost of volunteers among 18 wards ranged from yen 2,079 to yen 6,732, and those with the cost of volunteers ranged from yen 3,289 to yen 8,366. The net benefit per capita was negative. CONCLUSIONS: To conduct cost-benefit analyses of community based rehabilitation programs more precisely, it is necessary to measure the costs of medical care and long term care saved and the benefits to others (e.g. volunteers, community residents, etc.) and to assess the WTP so that various aspects of effectiveness (health and non-health benefits) are taken into account. PMID- 11868343 TI - [Identifying perceived needs among nurses in providing their patients with smoking cessation support in a Japanese hospital results from focus group interviews]. AB - OBJECTIVE: To identify perceived needs among nurses in providing their patients with smoking cessation support in a Japanese hospital. SUBJECTS: Thirty-three female nurses who were interested in providing their patients with smoking cessation support in the hospital setting participated in focus group interviews. They conducted six focus group interviews segmented by age, working conditions (e.g., outpatient or inpatient sections) and occupational class (e.g., administrative post or not). DESIGN: An experienced researcher moderated two focus groups and another researcher who received training moderated four focus groups. At least two observers hand-recorded participants' conversations in each group and all groups were also tape-recorded. Moderators and observers analyzed the focus group data together. RESULTS: We found that smoking cessation counseling or health education on effects of smoking to be rarely conducted in the hospital. The study participants ordinarily just told patients to stop smoking. They mentioned several barriers to conducting smoking cessation support and suggested a number of ideas to promote this aim in the hospital. CONCLUSIONS: The focus group interview is an effective means to determine the needs and interests of Japanese health professionals. Nurses have many opinions and insights for supporting smoking patients. To introduce a smoking cessation program into hospitals, there are various problems that should be solved. First, nurses and other health professionals should have accurate knowledge, positive attitudes and appropriate skills for smoking cessation support. Second, smoking cessation support should be programmed as part of treatment or nursing. Third, healthcare professionals, especially doctors, should collaborate in supporting patients to stop smoking. Fourth, the hospital environment should be modified to promote smoking cessation. Finally, hospitals should develop a consensus among all staff about the importance of smoking cessation support and smoking control activities. As the result, multidimensional strategies are needed to effectively promote smoking cessation support in the hospital setting. PMID- 11868344 TI - [The role and function of health science center in the community]. PMID- 11868345 TI - [Risk factor of nonadherence with tuberculosis therapy in Shinjuku-city's tuberculosis patients and directly observed therapy services]. PMID- 11868346 TI - [How can we prevent alveolar echinococcosis? Ecosystem and risk management viewpoints]. AB - PURPOSE: This article focuses on understanding epidemiological features of alveolar echinococcosis and discussing its prevention and control, especially from a viewpoint of the ecosystem and risk management. METHOD: Publications on alveolar echinococcosis throughout the world were systematically reviewed with special reference to ecology, epidemiology and countermeasures. RESULTS: Alveolar echinococcosis, caused by accidental infection with larva of the parasite Echinococcus multilocularis is fatal to humans unless diagnosed at an early stage. No effective control measures have been identified so far because it is difficult to fully understand the ecology of the parasite and its intermediate and definitive hosts. It is also not easy to determine the precise infection route to humans mainly because of the long latent period. In Hokkaido, infection rates among red foxes have recently risen even in low endemic districts. Not only stray and domestic dogs but also some pigs in Hokkaido have been found to be infected. While the number of reported human cases is still small, around 10 cases per year, local residents seem to be threatened with the risk of infection. DISCUSSION AND CONCLUSIONS: We predict that the incidence of alveolar echinococcosis among humans in Japan will increase in the near future if no effective preventive measures are conducted. In addition, Echinococcus multilocularis infection has the potential to affect the economy of Hokkaido because of its impact on the agricultural and tourist industries. Well-designed epidemiological surveys are therefore urgently required, in the context of ecosystem and risk management prior to large outbreaks. International collaboration is also desired. PMID- 11868347 TI - [Severe hyponatremia in a case of severe spinal injury: sequential examination of factors affecting water-electrolyte balance]. AB - Although hyponatremia has been known to occur in patients with severe spinal cord injury with highly incidence, its mechanism has not been understood well. We examined a 64-year-old patient with severe hyponatremia after spinal cord injury by sequential measuring of the factors affecting water-electrolyte balance, such as antidiuretic hormone, renin, angiotensin II, atrial natriuretic peptide, and brain natriuretic peptide. The patient showed severe hypotension due to dysfunction of the sympathetic nerve. The hyponatremia gradually resolved with the improvement of sympathetic nerve function. According to those results, the sympathetic nerve dysfunction was thought to correlate with the hyponatremia, and it was suggested that the unknown sympathetic regulation of water-electrolyte balance existed. PMID- 11868348 TI - [Juvenile-onset multiple brain infarcts localized in the posterior circulation: a case report]. AB - We report a 37-year-old male patient with multiple brain infarcts due to arterial lesions localized in the posterior circulation, who developed a paramedian pontine infarct on the left side. He had been treated as schizophrenia for 20 years. A cranial CT performed one year before showed old small infarcts in the territories of the bilateral thalamo-perforating and left thalamo-geniculate arteries and the right posterior inferior cerebellar artery. The vertebral and basilar arteries were small in diameter on MRI and MR angiography(MRA). Cerebral angiography revealed a narrow smooth basilar artery. In addition, the P2 segments of the bilateral posterior cerebral arteries were markedly narrow with irregular walls. Carotid arteriograms were normal and no atherosclerosis was found. The nature of these arterial lesions remains unknown in this case. Even if MRA shows vertebrobasilar artery hypoplasia, a known congenital risk factor of a posterior circulation infarct, we must rule out a possibility that some arterial pathology is going on. PMID- 11868349 TI - [Portal-systemic shunt with cerebellar ataxia as the initial neurological manifestation which was dramatically improved by an intravenous administration of branched amino acid]. AB - A 58-year-old woman began to show ataxia at age 45 and dysarthria at age 56. Neurological examination revealed slurred speech, truncal ataxia, and pyramydal sign. Neither history of alcoholism nor hereditary factors were found. The level of serum ammonia was increased. Brain MRI study showed a high signal intensity in the cerebral peduncle and globus pallidus and mild cerebellar atrophy on T1 weighted image. A portal-systemic shunt due to a shunt vessel was found between the left splenic and kidney veins although she did not show any other symptoms or signs due to liver cirrhosis. Her symptoms dramatically improved by an intravenous administration of branched amino acid. The present case suggests an importance in finding a treatable cerebellar ataxia. PMID- 11868350 TI - [A case of postcentral infarction presenting discriminative sensory impairment limited to the distal leg on one side]. AB - Presented is a case of postcentral infarction in a 70-year-old woman, which manifested as sensory disorders localized at the distal portion of the right lower extremity. Sensory disorders were characterized by the disturbances of discriminative sensation. Elemental senses were nearly normal. MRI revealed cerebral infarction localized at the superior portion of the postcentral gyrus and medial surface of the parietal lobe and the paracentral lobule of the left cerebral hemisphere. Carotid echography demonstrated stenotic lesions of the bilateral internal carotid arteries, suggesting that infarction of the cortical branch of the anterior cerebral artery due to artery-to-artery embolism was the cause of the condition. Cortical sensory disturbances due to lesions at the somatosensory area of the postcentral gyrus often appear on the upper extremities and face, and rarely occur as sensory disturbances that were localized in the distal portion of the lower extremities, as in this patient. The case suggests that it is important to keep in mind that sensory disorders that are localized in a single extremity with a distribution that suggests peripheral nerve lesions can be caused by cerebral cortical lesions. PMID- 11868351 TI - [Stereotyped movements in Wilson's disease: a case report]. AB - A 45-year-old Japanese man was admitted to our hospital because of persistent appearance of repetitive, purposeless stereotyped movements such as clapping the hands behind his back, scrubbing or rubbing movements of the hands. Laboratory examination showed low plasma copper and ceruloplasmin concentration. He was diagnosed as having Wilson's disease. Brain MRI revealed degeneration of the striatum and pallidum as well as atrophy of brainstem. Single photon emission computer tomographic scan (SPECT) showed hypoperfusion in the bilateral frontal lobe as well as basal ganglia. It is rare to see stereotyped movements in patients with Wilson's disease. Stereotyped movements observed in the present patient was not likely to be a simple stereotypy, since appearance of the motor acts was variable and complex. We consider that his stereotyped movement is a type of clonic perseveration induced by dysfunctioning of frontal cortico-basal ganglia-thalamo-cortical loops. PMID- 11868352 TI - [Multiple sclerosis--recent advances in pathogenesis and treatment]. PMID- 11868353 TI - [A case of FK 506-induced leukoencephalopathy]. AB - We reported a 15-year-old boy with an acute myelomonocytic leukemia and FK 506 induced leukoencephalopathy. He was received FK 506 for graft versus host disease occurred after peripheral blood stem cell transplantation. He, four weeks later, had generalized seizures and consciousness disturbance. The serum level of FK 506 was high (27.5 ng/ml). His brain MRI showed abnormal high intensity areas in the frontal and parietal white matter lesions on T2-weighted images. Neuropathological studies revealed the destruction of myelin sheeths and axons in the cerebral white matter corresponded with abnormal lesions on MRI. There were calcification and mineralization in the small vessel walls of the cortex and white matter. Osteopontin immunoreactivity was detected in the endothelial cells of small vessels. These findings suggest that the vascular damage was involved in the FK 506-induced leukoencephalopathy. PMID- 11868354 TI - [A case of the persistent vegetative state following symmetrical peduncular infarction]. AB - A 64-year-old woman presented with sudden onset of disequilibrium and dysarthria. Two days later she became lethargic and tetraparetic. An emergency MRI disclosed multiple infarctions in the medulla oblongata and pons. An angiogram revealed severe stenosis of the rostral basilar artery. The patient underwent a rapidly progressive course with impaired consciousness and flaccid tetraplegia. MR images 7 days after the onset disclosed a symmetrical midbrain infarction that was localized mainly in the cerebral peduncles, suggesting a distal basilar artery occlusion as the cause of this pathology. Since 4 weeks after the onset the patient was in the persistent vegetative state, which was essentially unchanged during the 1-year follow-up period. Persistent vegetative state is known to be caused by diffuse supratentorial or bilateral thalamic lesions, but in our case the lesions were restricted to the infratentorial area. We speculate a mechanism to cause the persistent vegetative state by partial impairment of the connection to the thalamic ascending system or extrathalamic ascending system, which leads to the dissociation of arousal(preserved in this case) and awareness(impaired in this case) after the ventral midbrain infarction. PMID- 11868355 TI - [MRI of the Ramsey Hunt syndrome: case report]. PMID- 11868356 TI - [Transient lesion in the splenium of the corpus callosum in a case with epilepsy]. PMID- 11868358 TI - [Reversible expression of CD34 by hematopoietic stem cells]. PMID- 11868357 TI - [Paraneoplastic neuropathy with respiratory failure preceded by depression]. PMID- 11868359 TI - [Lupus cystitis in the course of Evans syndrome]. AB - A 50-year-old woman with a 4-year history of Evans syndrome was admitted to our hospital because of progressive nausea, appetite loss, body weight loss, diarrhea and abdominal pain. Abdominal ultrasonography revealed pleural effusion, ascites, bilateral hydronephrosis, dilatation of the bilateral ureter, and irregular wall thickness of the urinary bladder. Immunological studies revealed decreased complement components (C3; 72 mg/dl, C4; 7 mg/dl, CH50; 28.8 mg/dl), a x 80 antinuclear antibody titer (homogeneous pattern), antibody against single stranded DNA 19 U/ml, anti-SS-A antibody over 500 U/ml and negativity for antibody against double-stranded DNA (anti-dsDNA Ab). Although the patient did not fulfill the criteria for systemic lupus erythematosus (SLE), we diagnosed her as having lupus cystitis. Bolus methylprednisolone (mPSL) therapy (1,000 mg mPSL over 3 days, div) was administered, followed by 60 mg PSL, and this led to immediate improvement of the patient's symptoms and laboratory data. Later, anti dsDNA Ab became positive, and the patient thereby fulfilled the criteria for SLE. Lupus cystitis following Evans syndrome has rarely been reported. The present such case was treated successfully with bolus mPSL therapy. PMID- 11868360 TI - [Superior mesenteric venous thrombosis during treatment of malignant lymphoma and of pure red cell aplasia]. AB - Mesenteric vein thrombosis is an uncommon type of intestinal ischemia that can be associated with significant mortality if its diagnosis is delayed. We experienced two patients with hematological disorders--non-Hodgkin's lymphoma (NHL) and pure red cell aplasia (PRCA)--who developed superior mesenteric vein (SMV) thrombosis during treatment. Neither of the patients had underlying disorders of the anticoagulant system that might have produced a hypercoagulable state. The first patient developed SMV thrombosis immediately after chemotherapy for NHL. This patient also had portal hypertension due to chronic hepatitis B. Direct injury to endothelial cells by the anti-cancer drugs and alteration of blood flow were the probable causes of the SMV thrombosis. The second patient with PRCA had regularly taken prednisolone, and this had induced a hypercoagulable state. The clinical symptoms of SMV thrombosis are usually non-specific, and in our patients vague, crampy abdominal pain without bloody diarrhea was the only complaint. Abdominal CT scan under a clinical suspicion of SMV thrombosis revealed the thrombi in the SMV. Urgent surgical resection of the infarcted bowel and immediate postoperative anticoagulation resulted in a favorable outcome. Clinicians should be aware of the vague symptoms of SMV thrombosis, as early diagnosis and urgent therapy are essential to prevent a fatal outcome. PMID- 11868361 TI - [Successful treatment with combined anti-thymocyte globulin and methylprednisolone for bronchiolitis obliterans after allogeneic bone marrow transplantation in a child with chronic myelogenous leukemia]. AB - A 9-year-old boy with secondary chronic myelogenous leukemia after treatment of acute lymphoblastic leukemia underwent allogeneic bone marrow transplantation (BMT) from an HLA-identical sibling in December 1998. Grade II acute GVHD developed on day 24 and chronic GVHD developed 5 months after BMT. Cough and dyspnea appeared 9 months after BMT. Despite administration of tacrolimus and methylprednisolone (m-PSL) pulse therapy, the dyspnea gradually increased in severity. Bronchiolitis obliterans was diagnosed on the basis of lung biopsy in January 1999. Because renal dysfunction made it difficult to continue the use of tacrolimus, we attempted antithymocyte globulin (ATG) + m-PSL therapy. Major BCR/ABL mRNA was transiently positive on RT-PCR after the ATG + m-PSL therapy, but no severe complications were observed. A decreasing V50/V25 level and increasing peak flow value were observed in respiratory function tests after ATG + m-PSL therapy, and the patient is currently free of dyspnea. Our findings suggest that ATG + m-PSL therapy is beneficIal for patients with drug-resistant bronchiolitis obliterans after BMT. PMID- 11868362 TI - [Mixed warm and cold antibody-type autoimmune hemolytic anemia in a healthy pregnant woman with primary cytomegalovirus infection]. AB - A 25-year-old previously healthy pregnant woman was admitted to our hospital because of severe anemia (Hb 6.7 g/dl), and diagnosed as having mixed-type autoimmune hemolytic anemia (AIHA) due to de novo cytomegalovirus (CMV) infection. After daily administration of prednisolone, the anemia gradually resolved and the patient delivered a healthy baby. This is the first report of a healthy person suffering from mixed-type AIHA due to de novo CMV infection. PMID- 11868363 TI - [Plasma exchange and continuous hemodiafiltration as an initial treatment for diffuse large B-cell lymphoma-associated hemophagocytic syndrome]. AB - A 68-year-old man was admitted to our hospital because of fever, jaundice and hepatosplenomegaly. A diagnosis of diffuse large cell, B-cell type malignant lymphoma, associated with hemophagocytic syndrome (LAHS), was made. CT scan revealed lymphadenopathy in the abdominal cavity and multiple tumors in the spleen. Performance status and hepatic coma grade were 4 and II, respectively. Laboratory findings showed bicytopenia (Hb 9.9 g/dl, platelet 35 x 10(3)/microliter), severe liver dysfunction (ALP 1,115 U/l, gamma-GTP 437 U/l, T.Bil 15.4 mg/dl, D.Bil 12.8 mg/dl) and elevated levels of beta 2 microglobulin (12.9 mg/dl), ferritin (2,300 ng/ml) and sIL-2 receptor (36,900 U/ml). Plasma exchange (PE) and continuous hemodiafiltration (CHDF) enabled the patient to undergo diagnostic procedures, irradiation (total 34 Gy) and chemotherapy. Biopsy specimens revealed infiltration of lymphoma cells into the liver and bone marrow. We measured the blood concentrations of TNF-alpha, IL-6, and IL-8 before and after PE and CHDF by the ELISA method, and found normalization of hypercytokinemia after the procedure. It was suggested that initial treatment with PE and CHDF was effective for control of HPS, enabling us to perform chemotherapy for the lymphoma. PMID- 11868365 TI - [Clinicopathologic features of intravascular large B-cell lymphoma in Japan: review of the special reference to the Asian variant]. AB - The Asian variant of intravascular large B-cell lymphoma (IVL) is characterized by hemophagocytic syndrome, i.e. the clinical features include pancytopenia, hepatosplenomegaly, and rarely mass formation. It usually lacks any neurological abnormality or skin lesions, which are typical features of classical IVL. Sixty seven cases of IVL reported in Japan since 1990 were classified into two groups, and their clinicopathologic features were compared. Forty-five cases of IVL that met the clinical and laboratory criteria for the Asian variant were classified into Group A (average: 66 yr; male: 62%), and 22 cases that did not meet the criteria were classified into Group C (average: 65 yr; male: 73%). Hemophagocytosis was described only in Group A. The positivity rates for bone marrow invasion, fever, hyperbilirubinemia and elevated levels of LDH and CRP were significantly (p = 0.0037, p < 0.0001, p = 0.0428, p = 0.0108 and p = 0.0008, respectively) higher in Group A than in Group C. On the other hand, the positivity rates for neurological abnormality and skin lesions were significantly (p < 0.0001 and p = 0.0011, respectively) higher in Group C than in Group A. Our reassessment of the reported cases of IVL in Japan identified two major categories of IVL: the classical form and the Asian variant. PMID- 11868364 TI - [Acute myelogenous leukemia associated with severe esophageal stricture after chemotherapy]. AB - A 41-year-old woman with relapsed acute myelogenous leukemia was treated twice with idarubicin hydrochloride and cytarabine. She developed a 6-cm-long stricture in the lower esophagus 12 days after re-induction therapy. Although she had preceding candida infection, it is suspected that her stricture was caused by mucosal damage due to chemotherapeutic agents. This case suggests that the possibility of esophageal stricture should be considered in patients with swallowing disturbance after intensive chemotherapy. PMID- 11868366 TI - [From a view of the wheelchair users--a training report of the students on barrier-free environment in Sapporo]. AB - This short report describes what we learned from our training session on the use of the wheelchair in the various parts of Sapporo City. The session was conducted in July 2000 as one of the public health field training programs for fourth-year medical students of Hokkaido University School of Medicine. We, as a set of 5 people with the wheelchair, actually toured the street, subways, big department stores, banks, and other public facilities and buildings to assess their social and physical environment in a context of barrier-free society, so that some suggestions may be given to the City for the development of an appropriate welfare system. We also evaluated people's attitudes to the wheelchair users. We learned that 1) it is not easy for the wheelchair users to move around; it's much more difficult than we thought; 2) there are a lot of barriers and obstacles in a social environment for the wheelchair users; 3) many people have not noticed these barriers. This training was very instrumental in terms of understanding the barrier-free environment from the view of wheelchair users. Comprehensive city planning is necessary to accommodate the various individuals and senior citizens with different handicaps. Furthermore, our own view of a barrier-free society must be changed. PMID- 11868367 TI - [Human genome and made-to-order health care system]. AB - Common diseases of adulthood are caused by interplay between environmental and genetic factors. Essential hypertension, diabetes mellitus, and coronary heart disease are the representative diseases, being the major cause of disability in aged population. The molecular pathogenesis of the most common forms of the diseases is still poorly understood. However, thanks to the rapid progress of "Human Genome Project", susceptibility genes of common diseases are expected to be isolated in the near future. At present, the word, "made-to-order medicine", means personalized medication in terms of doses or drug groups determined by genotypes of drug metabolizing genes. However, if knowledge about susceptibility genes and specific mutations for common diseases is available, made-to-order health care would be feasible. Before attainment of made-to-order health care system, we have to reach social agreement in usage of genetic informations. Genetic information can be used to prevent the onset of diseases, to assure early detection and treatment. This information can be used by probands and their relatives, but also by third parties such as insurers and employers. There are concerns that this information could lead to discrimination and social exclusion. PMID- 11868368 TI - [Oleylamine (long-chain fatty amine)-induced cell death through MAP kinase pathways in human pancreatic cancer cells]. AB - We have previously shown that farnesylamine which acts as a farnesyltransferase inhibitor (FTI) induces apoptosis in human pancreatic cancer cells. Since the effect of FTI is usually known as "cytostatic", another apoptotic machinery of farnesylamine should be revealed in addition to the inhibition of farnesylation. Considering the chemical formula of farnesylamine, the "long-chain fatty amine (LFA)" structure may have a critical role in this mechanism. In this experiment, we examined the cytotoxic effect of LFA with alkyl-chain including oleylamine. Exposure of human pancreatic cancer cells to LFAs resulted in cell death, whereas short-chain fatty amine and oleylamine-related compounds without "amine" did not exert cytotoxicity. Oleylamine-induced cell death was confirmed as apoptosis by DNA laddering and caspase-dependent, but numerous cytoplasmic vacuoles, a typical feature of autophagy (type-2 cell death), were also observed. Preceding the apoptosis, strong and sustained activation of c-jun N-terminal kinase (JNK) and p38 was observed; caspase inhibitors did not attenuate activities of those kinases despite significant inhibition of apoptosis. Blockage of JNK activity by dominant-negative mutant completely abrogated the oleylamine-induced DNA laddering, but not autophagy. Furthermore, oleylamine decreased extracellular signal-regulated kinase (ERK) activity, probably through the activation of mitogen-activated protein kinase phosphatase-1. Taken together, LFA induces apoptosis through activation of JNK, p38 and caspase, accompanied with ERK inactivation, in human pancreatic cancer cells. PMID- 11868369 TI - [Recent advances in innate immune host defense--hose defense lectins and endotoxin receptors]. AB - Accumulating evidence has revealed that CD14 and Toll-like receptors (TLRs) function as pattern recognition receptors for broad ranges of pathogens. The structural characteristics of these proteins is that they possess leucine-rich repeats, which appear to be involved in protein-protein interaction. We have shown that the extracellular TLR2 domain directly binds to peptidoglycan (PGN) which is abundant in the cell wall of Gram-positive bacteria and that the N terminal region of TLR2 is critical for the recognition of PGN. Surfactant proteins A and D (pulmonary collectins) function as host defense lectins and play important roles in innate immunity in the lung-Collectins including SP-A, SP-D and mannose-binding proteins can bind CD14. We have demonstrated that SP-A modulates the cellular response to smooth and rough lipopolysaccharides (LPS) by interaction with CD14. In addition, this protein has been revealed to function as a negative regulator against PGN-induced TNF-alpha secretion from macrophages through direct interaction with TLR2. We propose that SP-A modulates inflammatory responses against the bacterial components by interaction with pattern recognition receptors. PMID- 11868370 TI - [Purkinje cells of the monkey vermal lobe: simple-spike activity during pursuit and passive whole body rotation]. PMID- 11868371 TI - [Renal hemodynamic abnormalities in spontaneously diabetic rat resembling to human type 2 diabetes mellitus]. PMID- 11868372 TI - [Pharmacological analysis of inhibitory effect of sarpogrelate hydrochloride on human radial artery contraction]. PMID- 11868373 TI - [Mechanism of liver-specific reduced expression of glutathione peroxidase in LEC rat]. PMID- 11868374 TI - [Protective effect of nicorandil (ATP-sensitive potassium channel opener) on ischemic rabbit spinal cord]. PMID- 11868375 TI - [The past and the future prospects of acute medicine]. AB - Recent development and the future prospects of acute medicine as well as emergency medical treatment system in Japan are reviewed. At first, I described the history of medical system, transporting system, and information system in the emergency medical treatment system, and then strongly emphasized the practical use of this system. The base of the system was constructed in the middle 1970s and has greatly developed during recent two decades. The development of the emergency medical treatment system could let us know an important role of the education of acute medicine for medical students in the university. Now a lot of medical student can learn acute medicine at university in our country. The education of acute medicine should be expanded to nurses, comedical stuffs, ambulant teams, and the citizens who are the constituent member of the prehospital care. Based on the establishment of emergency medical treatment system and the spread of acute medicine, I believe that Japanese people can get an affluent and safety life. I hope more development of emergency medical treatment system and acute medicine in the 21st century. PMID- 11868376 TI - [Analysis of JC virus early antigen in transformation and its implication in human medulloblastoma]. PMID- 11868377 TI - [Functional and urodynamic characteristics in patients with ileal neobladder]. AB - We analyzed the functional and urodynamic characteristics in 19 patients with ileal neobladder by the Hautmann procedure. A questionnaire survey by mail was performed for functional information of neobladder. Seventeen of the 19 patients (89.5%) could voluntarily void via the urethra and the others needed clean intermittent self catheterization (CIC) because of their significant residual volume. Eight of the 19 patients (42.1%) micturated at least two times at night. Two of the 19 patients (10.5%) were incontinent in the day time and 12 (63.2%) in the night time. They needed 2 pads in the day time and one pad at night on average. Eight out of 18 patients (44.4%) were satisfied with their micturition state. A urodynamic study showed the neobladder to be a low-pressure reservoir with a mean capacity of 395.2 +/- 96.8 ml. The mean residual volume of the patients without CIC was 27.8 +/- 28.2 ml. In 10 out of 11 patients high frequency and high amplitude spikes were seen by the perineal electromyogram in the voiding phase. PMID- 11868379 TI - [48XXYY Klinefelter's syndrome with recurrent foot ulcers: a case report]. AB - A case of 48XXYY Klinefelter's syndrome diagnosed with foot ulcers, in a 43-year old man, is reported. He presented to the Department of Dermatology in our hospital with the chief complaint of recurrent foot ulcers. He was referred to us because of atrophic external genitalia. The present condition and endocrinological examination suggested Klinefelter's syndrome. Chromosomal analysis revealed that his chromosomal type is 48XXYY. This is the 24th case of Klinefelter's syndrome with 48XXYY chromosome and is the 5th case associated with foot ulcers in Japan. Here we report this case together with a brief review of the previously reported cases. PMID- 11868380 TI - [Cholinergic crisis following administration of distigmine bromide: a case report]. AB - The long-acting anticholinesterase, distigmine bromide (Ubretid), is widely used for the treatment of underactive neurogenic bladder. Therefore, we emphasize its hazardable side-effect of cholinergic crisis. A 78-year-old man with duodenal ulcer complained of nocturia, and was administered distigmine bromide 10 mg daily under the diagnosis of mild benign prostatic hypertrophy with underactive neurogenic bladder. It seemed that administration slightly improved his symptom but he developed bradycardia, dyspnea and drowsiness suddenly on the 4th day. Blood examination revealed extremely low cholinesterase in his serum, suggesting distigmine bromide intoxication. He was treated intensively with several intravenous injections of atropine, high-concentration oxygen and transfusion of fresh frozen plasma. Nevertheless, his condition did not recover, resulting in death of "cholinergic crisis" on the 6th day. PMID- 11868378 TI - [Clinical study on prostatic cancer detected incidentally by transurethral resection of the prostate]. AB - Transurethral resection of the prostate (TUR-P) was performed on 463 consecutive patients with clinically diagnosed benign prostatic hyperplasia (BPH) between April 1994 and June 2000. Pathological examinations of resected prostatic tissues revealed prostatic cancer in 15 (3.2%) of them. Eight (53.3%) of them were in stage A1, and 7 (46.7%) in stage A2. Between 15 cases with prostatic cancer and those with BPH, clinical features including age, serum prostate specific antigen (PSA) levels, prostatic volume, PSA density (PSAD), and resected prostatic tissue weight were compared. As a result, age was the only parameter related with prostatic cancer with a statistically significant difference. The higher the age, prostate cancer was found more frequently. Postoperatively, radical prostatectomy and antiandrogen therapy were performed in 1 and 9 cases, respectively. The remaining 5 cases are being followed with no treatment for prostatic cancer, and have shown no findings suggesting recurrence. These 15 cases are all living disease-free at present. It seems of importance to explain preoperatively the possible detection of prostatic cancer in association with TUR-P, particularly for elderly patients aged 80 years or older. PMID- 11868381 TI - Retroperitoneoscopic ureteroureterostomy for retrocaval ureter. AB - A 10-year-old boy presented to our hospital with pain in the flank and was subsequently diagnosed as having a retrocaval ureter. He underwent retroperitoneoscopic surgery, during which the retrocaval segment of the right ureter was divided and reanastomosed anterior to the inferior vena cava using 5-0 polyglactin. He returned to normal activities from three days after the procedure. Hydronephrosis was markedly decreased on excretory urography at 6 months after surgery. To the best of our knowledge, this is the eighth case report on retroperitoneoscopic surgery for retrocaval ureter in the English and Japanese literature. PMID- 11868382 TI - [Carcinosarcoma of the renal pelvis and ureter: a case report]. AB - We report a case of carcinosarcoma of the renal pelvis and ureter arising in an 89-year-old man who presented at our hospital with gross hematuria. Abdominal computed tomography, excretory pyelography, and retrograde pyelography demonstrated that left hydronephrosis was caused by an ureteral tumor. Left urine cytology indicated transitional cell carcinoma. The patient underwent chemotherapy and radiation therapy. However, gross hematuria recurred, and the patient underwent left nephroureterectomy. The surgical specimen showed carcinosarcoma in the renal pelvis and ureter histologically. He has been free of cancer for 1.5 years. PMID- 11868383 TI - [A case of regionally metastatic pure squamous cell carcinoma of the urinary bladder successfully treated with radical chemoradiotherapy]. AB - We report a case of regionally metastatic pure squamous cell carcinoma of the urinary bladder successfully treated with combined radiation and chemotherapy in a 46-year-old man. Clinical staging was T3bN2M0. The patient received 50 Gy external radiation combined with intraarterial and systemic chemotherapy. Pathological complete response was found both in bladder and regional lymph nodes when he underwent radical cystectomy and lymph node dissection. The patient has been alive without evidence of disease for two years postoperatively. PMID- 11868384 TI - [Scrotal emphysema: a case report]. AB - We report a case of scrotal emphysema following the placement of a chest tube and an endotrachial catheter for traumatic pneumothorax. Scrotal emphysema is a rare condition and is considered as a type of pneumoscrotum, for which 4 possible mechanisms have been discussed. It is important to promptly determine the origin of air because this condition may be caused by a life-threatening disease. PMID- 11868385 TI - [The 90th Annual Meeting of Japanese Urological Association]. PMID- 11868386 TI - [A comparative study assessing clinical effects of naftopidil and tamsulosin hydrochloride on benign prostatic hyperplasia]. AB - We compared the efficacy of naftopidil with that of tamsulosin hydrochloride for benign prostatic hyperplasia patients. Eighty-five patients without improvement of quality of life (QOL) score by the administration of 50-75 mg naftopidil for more than four weeks were assigned to receive doses of 0.1-0.2 mg tamsulosin hydrochloride and 89 patients without improvement of QOL score by the administration of 0.1-0.2 mg tamsulosin hydrochloride for more than four weeks were assigned to receive doses of 50-75 mg naftopidil once a day for 8 weeks. International prostate symptom score, maximum flow rate, residual urine volume and side effect profile were determined before the administration of the first medicine, before the administration of the second medicine and after 8 weeks of treatment with the second medicine. In the group without improvement of QOL score by naftopidil, significant improvements in symptoms of urgency, weak stream and straining were observed after 8 weeks of treatment with tamsulosin hydrochloride. In the group without improvement of QOL score by tamsulosin hydrochloride, significant improvements in symptoms of incomplete emptying, intermittency and nocturia were observed after 8 weeks of treatment with naftopidil. In conclusion, improvement of symptoms by each alpha 1-blocker differs symptom by symptom. Tamsulosin hydrochloride was superior to naftopidil for the symptoms of urine flow and naftopidil was superior to tamsulosin hydrochloride for the symptom of nocturia. PMID- 11868388 TI - [Proteins in response to environmental stress]. AB - Last decade, remarkable progress was achieved in the field of environmental sciences, especially those related to biological functions against environmental stress. Recent progress has been focused on the transcriptional control of proteins, such as cytochrome P450, UDP-glucronosyltransferase, and metallothionein. Along with these studies, entirely new functions of some proteins were identified; for example heme oxygenase-1, originally isolated by the catalytic activity of heme, is nowadays recognized as one of the stress proteins. We are, therefore, interested in the relationship among proteins of body defense mechanisms; i.e., co-regulation of two (or more) distinct proteins, and why they should be under the same regulation. One possible idea is that they are members of the defense system against active oxygen, which will be described by Drs. Sato and Inoue in a future review. Alternatively, they might have evolved from an ancestor of the defense system that originally protected from oxygen toxicity. Findings on AhR/Arnt and HIF1 systems demonstrating that they are members of the same PAS family seem to support this hypothesis. One of the missing links of the hypothesis is the so called oxygen sensor, suggesting (a) hemoprotein in 1988. A review by Dr. Ogawa on Bach1, the first vertebrate transcription factor controlled by heme, will provide on answers to the missing links. Through this series of reviews, we have a rough map of the biological system against environmental stress to explore the new frontier of environmental science. PMID- 11868387 TI - [Guided bone regeneration around dental implants using a bioabsorbable membrane. A pilot investigation in experimental animals]. AB - The aim of this experimental investigation was to evaluate the effect of guided bone regeneration around immediately placed implants using different barrier membranes. Five adult fox hounds were used in this investigation. After extraction of all premolars in the lower jaw, implant osteotomies in the regions of the former premolars and additional buccal bone defects (5 mm x 5 mm) were created. Subsequently, the implants were placed. The defects were treated with one of the following three modalities: (a) guided bone regeneration, using a bioinert expanded polytetrafluoroethylene membrane, (b) no membrane application and (c) guided bone regeneration, using a bioabsorbable membrane made from a synthetic copolymer of lactide and glycolide. After a healing period of six months, the animals were sacrificed and the implants with the surrounding tissues processed for histologic evaluation. The clinical pretreatment defects between the different treatment groups were not statistically different (bioinert membrane group: 4.8 mm; control group: 4.3 mm; bioabsorbable membrane group: 4.9 mm). The remaining histological defects after a healing period of 6 months amounted to 3.2 mm for the nonresorbable group, to 5.6 mm for the control and to 6.3 mm for the bioabsorbable group. A significant difference was observed between the bioinert membrane group and the other two groups. The mineralized bone-to implant contact in the bioinert membrane group was 52%, in the control group 47% and in the bioabsorbable membrane group 43.3%. The values were not statistically significant different. The results of this study indicate that a partial bone regeneration with bioinert e-PTFE membranes around immediately placed implants is possible. The utilized bioabsorbable polylactide/polyglycolide membrane did not show any bone regenerative effect, and the results did not differ from the control group without membrane application. PMID- 11868389 TI - [Reactive oxygen]. AB - Mammalian tissues have large amounts of available ATP which are generated by oxidative phosphorylation in mitochondria. To maintain the body, large amounts of oxygen are required to regenerate the ATP molecules. A small fraction of the inspired oxygen is converted to superoxide radicals and related metabolites even under physiological conditions. Most reactive oxygen species react rapidly with a variety of molecules, thereby interfering with cellular functions and various diseases. PMID- 11868391 TI - [Regulation of cytochrome P-450 (CYP) genes by nuclear receptors]. AB - Many xenobiotics are metabolized and detoxified by cytochrome P-450s (CYP). The xenobiotics metabolizing CYPs are induced by various kinds of receptors. To induce CYP1A1, the Ah receptor requires a ligand for its activation as a transcription factor. On the other hand, benzimidazole compounds can induce CYP1A1 without binding to the Ah receptor as a ligand (ligand-independent pathway). In response to phenobarbital (PB) and other PB-type inducers, the nuclear receptor CAR (the NR-constitutive active receptor) translocates to the nucleus, forms a dimer with the retinoid X receptor (RXR), and activates the PB responsive enhancer module (PBREM) in the PB-inducible CYP2B genes. For human CYP3A4 genes, pregnane X receptor (PXR) binds to the xenobiotic-responsive enhancer module (XREM) and upon induction by rifampicin, a PXR:RXR heterodimer could transactivate XREM. PMID- 11868390 TI - [Heme metabolism in stress response]. AB - Heme and its metabolism fulfill significant roles in many homeostatic and adaptative reactions. For example, heme (protein) senses oxygen concentration to regulate hypoxia response genes such as erythropoietin, and free heme, a proxidant, controls levels of several oxidative stress response proteins as well as that of a few enzymes in the heme metabolic pathway. Heme oxygenase (HO) is the key enzyme in heme catabolism, which degrades heme to Fe, CO, and biliverdin. CO is known as a gaseous messenger in the vascular and nervous systems. Biliverdin is rapidly converted to bilirubin, whose antioxidative effect is proposed to protect cells against reactive oxygen species. HO-1, the inducible isozyme of HO, is induced not only by its substrate heme, but also NO, metals, hypoxia, and various other stimuli. Studies on HO-1 deficiency indicate that induction of HO-1 is essential to homeostasis, at least in humans. Heme response elements (HREs), which mediate the induction of HO-1 expression by heme, are identified in enhancer regions of the mouse HO-1 gene. HRE shares a nucleotide sequence with the Maf recognition element (MARE). A transcriptional activator, Nrf2, has been shown to participate in HO-1 induction by several stimuli, including heme via HRE. A heme-binding transcription factor such as yeast Hap1 had been supposed to also exist in vertebrates, however, no such factor had been identified. Recently, we found that a mammalian transcription repressor, Bach1, directly binds heme, and that the DNA binding activity of Bach1 is negatively regulated by heme. Bach1 is capable of competing the binding to MARE with activators including Nrf2, therefore, HO-1 and other stress response genes bearing MARE may be regulated by heme via the MARE-binding transcription factors. Further analyses on the gene regulatory mechanism by heme would help us to understand the stress response system, especially against oxidative stress. PMID- 11868392 TI - [UDP-glucuronosyltransferase]. AB - UDP-glucuronosyltransferases (UGTs) represent a family of enzymes that glucuronidate many internal substances and drugs. This family acts as a drug metabolism phase II reactor in the liver and comprises one of the major protective mechanisms from toxic chemical substances. UGTs have two subfamilies; UGT1 and UGT2. UGT1 gene expresses 12 isoforms which are produced from a single gene by alternative splicing of a primary transcript. Each isoform has specific substrates. UGT1A1 conjugates bilirubin, and mutations of the gene cause hereditary unconjugated hyperbilirubinemias (Crigler-Najjar syndrome and Gilbert's syndrome). Recently, polymorphisms of UGT1A1 were revealed. In the Japanese population, there is a polymorphism of G71R and this mutation is a risk factor of neonatal hyperbiliruibnemia and a genetic cause of breast milk jaundice. These polymorphisms of UGTs might contribute to individual variations of drug metabolism and toxicity as well as inherited diseases. PMID- 11868393 TI - [Protective stress responsive protein: metallothionein]. AB - Metallothionein (MT) is a small, cysteine-rich, metal-binding protein. MT synthesis is induced by various stimuli such as heavy metals, oxidative stress, anticancer drugs and fasting stress. MT is capable of not only reducing metal toxicity but also scavenging free radicals. In fact, MT is involved in the protection of tissues against various forms of oxidative injury, including radiation, lipid peroxidation, oxidative stress caused by anticancer drugs, and conditions of hyperoxia. However, MT still lacks a manifest established biological function. Recently, transgenic mice with loss-of-function mutations in the MT-I/-II genes were established. Unexpectedly, the mice were in apparent good health, and the critical biological roles for MT have been questioned. Here the basic characteristics of MT are reviewed, the current MT study highlights summarized, and the putative biological functions of MT discussed. PMID- 11868394 TI - [Keshan disease--a review from the aspect of history and etiology]. AB - OBJECTIVE: Keshan disease is an endemic cardiomyopathy found in Keshan, north east China. The first patient was identified in 1935. This disease is characterized by a blood circulation disorder, endocardium abnormality and myocardium necrosis. Selenium (Se) deficiency is thought to be a major factor by Chinese scientists. However, the exact etiology has not been clarified up to now. The government decided to apply sodium selenite to growing crops, and the incidence of the disease decreased dramatically. However, a few cases still occur as chronic or latent types. This paper reviews Keshan disease from a historical aspect and also the present situation. METHODS: We made a reference survey and summarised the etiology, pathological changes, clinical manifestation, and other views of Keshan disease. RESULT: So far, epidemiological surveys have shown that Keshan disease occurs in large areas in 14 provinces in China, mainly in the countryside. It has been confirmed by clinical and pathological studies that Keshan disease is an independent endemic myocardial disease caused by biological and geological factors. The largest prevalence age rates are boys under 15 years old and women of childbearing age. There are several hypotheses; acute carbon monoxide poisoning, virus infection, malnutrition, or selenium deficiency. The first is not currently believed to be the cause. The following was pointed out; studies on the relationship between diet and the endemic areas of Keshan disease in 1961, where the food custom of the local residents was relatively simple and a so-called "one-sided diet" (eating a limited variety of food) might be related to the incidence of Keshan disease. In 1973, large-scale investigations on the natural environments were performed in the endemic areas of Keshan disease in the whole country. As a result, it was reported that there was a relationship between the incidence of Keshan disease and the special natural environment in the endemic areas and the cause of Keshan disease was strongly supported by nutritional, biological, geological and chemical (selenium deficiency) factors. In 1981, on the other hand, it was found that the levels of antibodies against Coxsackie virus were higher in the serum of Keshan disease patients than of normal subjects. This fact supposed that the cause of Keshan disease was related to a virus infection. However, it is difficult to explain why the clinical and pathological manifestations of Keshan disease are similar to those of other diseases, e.g. idiopathic dilatational myocardial disease. Further research should be performed on Keshan disease to clarify the etiology. PMID- 11868395 TI - [Bibliographical study of Minamata disease]. AB - In 1958, Minamata Disease was suggested to be organic mercury compounds intoxication. This suggestion was based on Hunter and Russel's report on occupational exposure to methylmercury. This report is known to have established the typical symptoms of methyl mercury intoxication. However, it has been widely believed since the official recognition of Minamata Disease (1956) that, at the moment of outbreak, no reports were available on organic mercury formation from inorganic mercury in acetaldehyde production from acetylene, or on organic mercury intoxication among workers in acetaldehyde production from acetylene. However, this was not the case. The formation of organic mercury from inorganic mercury used as a catalyst was reported by Vogt and Nieuwland in 1921. In 1930, Zangger reported several cases of organic mercury intoxication among workers in acetaldehyde production from acetylene. Soon after, Koelsch reported that the cases were methyl- and/or ethylmercury intoxication, and that such cases had been common since 1916. These reports were already available at the time of the Minamata Disease outbreak. However, Zangger's report, the most important of these three was not referred to until 1987, notwithstanding its listing in the references of Kurland et al.. Zangger's report was not referred to not by investigators, but by a lawyer. If these reports had been referred to at the outbreak of Minamata Disease, the number of victims in Minamata would have been minimized, and Minamata Disease in Niigata would have been prevented. PMID- 11868396 TI - [Gender and relation of life-style to morale in older people living in regional cities]. AB - The purpose of this study was to examine the morale of older people living in 7 cities (and towns), using the PGC morale scale, from the relationship between morale score and life-style. Data were collected from 1,269 people aged 60 or more in the community (619 males and 650 females) in cities (and towns). A questionnaire included the PGC morale scale and 14 life-style factors. There was no remarkable gender difference in the morale score. The relationship between morale and life-style were different in both sexes and higher in males. Female morale when living with a husband is generally high. It is important for females to have a husband. In the relationship between economics and morale, economic satisfaction was considered to be more important than the level of income. Eating regularly, participating as a volunteer and having a best friend are related to morale, but factors of job, smoking, drinking and sports enforcement-frequency are not. It is considered that there is no significant gender difference, but the relationship between morale and life-style in older people living in cities (towns) are different in both sexes. PMID- 11868397 TI - [A sexual health study of high school students at the 3 high schools in Kitakyushu City]. AB - OBJECTIVES: The purpose of this study was to clarify the actual sexual behavior and attitudes of high school students in Kitakyushu city, Fukuoka and then to develop effective sex education methods for high school students in this region. METHODS: This study investigated the sexual behavior and attitudes of 1,297 high school students in Kitakyushu by self-administered questionnaire. The differences in their answers by sex, prevalence of sexual intercourse and change in sexual behavior and attitude before and after the sex education lecture were examined. RESULT: 39.3% of the students had had sexual intercourse and 74.1% answered that they might have sex, if it were with a partner whom they loved. However, they did not have enough knowledge about contraception and sexually transmitted diseases. This result shows that they did not recognize the risks accompanying sexual intercourse. There are significant differences between male and female students in their sexual attitudes. Male students tend to permit premarital sexual intercourse, unfaithfulness, prostitution, hiring a prostitute and abortion. Male students tend to give more approval to the following opinions: both men and women should agree to sexual contact if the partner wants it; men should take the initiative in sexual contact; women should not talk about sex. Many female students answered that women should make their own decisions to have or not to have sex, however a considerable number of female students answered that for their first intercourse, they just agreed with their partner even though they really did not want to do so. After the sex education lecture, the students have more knowledge about contraception and STDs. However, there is no significant difference in their sexual attitudes before and after the lecture. CONCLUSION: In order to facilitate more desirable and safer sexual behavior among the younger generation, it is not enough to simply give them knowledge about contraception or STDs, etc. To organize more comprehensive sex education, it is also important to pay enough attention to gender problems and other social factors such as family background or regional background, etc. PMID- 11868399 TI - [Gender and age-related differences in life style characteristics, health condition, and unidentified complaints in healthy old-aged people]. AB - PURPOSE: The purpose of this study was to examine gender and age-related differences in life style such as exercise and dietary habits etc., health conditions and unidentified complaints in healthy older-aged people. METHOD: A questionnaire consisting of 40 items on life style and 16 items on unidentified complaints was administered to 890 people aged 60 to 89 in the community (304 males and 586 females). The proportions of gender and age-range for each item were computed, and a two-way analysis of variance based on the Lancaster method was applied. RESULTS: Most of the aged spend their time living regularly and have leisure activities. The smoking and drinking rates for males are higher than those for females in all age-ranges. The aged exercise more than one or two days a week for both sexes. Many aged males have exercised more than three days a week and/or for more than seven years. Most of the aged have regular dietary habits and there are many males who are careful to eat appropriate portion sizes. Females have more interest in food in the higher age-ranges, but tend to control meal-portions voluntarily. The rate of seeing a doctor regularly and having fractures increases in the higher age-ranges. The rates of arthritis and neuralgia are higher for females than for males. More females complain of 'eyes have grown blurred' and 'pain and swelling in a joint' and males complain more of 'cough and phlegm' and 'feel very thirsty'. CONCLUSION: Gender and age-related differences exist in most life style items for healthy old-aged people, and there is little in the way of age-related differences in the content of unidentified complaints. PMID- 11868400 TI - Addressing the needs of women living with Schizophrenia. PMID- 11868398 TI - [The relationship between undergoing a basic health checkup based on the Health and Medical Services Law for the Aged and medical expenditure covered under the National Health Insurance in Japan]. AB - OBJECTIVES: This study examined the relationship between undergoing the basic health checkup based on the Health and Medical Services Law for the Aged among urban residents and medical expenditure covered under the National Health Insurance in Japan. METHODS: The subjects were all residents of a city with a population of three hundred and fifty thousand, covered by National Health Insurance and over forty years of age. Medical expenditure was compared by age and sex between examinees who had undergone the basic health checkup for the past three years running and non-examinees who had had no checkup. In addition, from the results of a questionnaire survey of randomly sampled citizens, we compared the living habits of the above two groups, the examinees and non-examinees. RESULTS: For outpatients, the medical expenditure per case, the medical expenditure per capita for seniors, and the medical expenditure per day were lower, and the number of examination days per case and the examination rate (the number of receipts per capita) were higher in the examinee group than in the non examinee group. For inpatients, the medical expenditure per case, the medical expenditure per capita, the medical expenditure per day for seniors, the number of examination days per case, and the examination rate were lower in the examinee group compared to the non-examinee group. The increase in medical expenditure with age was less in the examinee group than in the non-examinee group. In men, the examinees had significantly better living habits (especially exercise and meals) than the non-examinees. No differences in living habits were found in women. CONCLUSIONS: Medical expenditure was lower among examinees than non examinees. In men, regular examinees had better living habits than non-examinees. PMID- 11868401 TI - A native smudging ceremony. PMID- 11868402 TI - Women's perceptions of breast cancer screening and education opportunities in Canada. PMID- 11868403 TI - The experiences of returning P.E.I. nurses. PMID- 11868405 TI - The gift of life. PMID- 11868404 TI - Come on nurses: wash your hands! PMID- 11868408 TI - Interview with Ginette Lemire Rodger. Interview by Marla Fletcher. PMID- 11868406 TI - Addressing overload practices. PMID- 11868409 TI - Public perceptions of and comfort level with nurse practitioners in family practice. PMID- 11868407 TI - Getting comfortable with research. PMID- 11868410 TI - Dietary intervention. Bowel care and maintenance in long-term care. PMID- 11868411 TI - Professional liability during the shortage. PMID- 11868412 TI - Making mandatory education fun. PMID- 11868414 TI - Narrative as bioethics: the "fact" of social selves and the function of consensus. PMID- 11868415 TI - Two models of ethical consensus, or what good is a bunch of bioethicists? PMID- 11868417 TI - Keeping company: ethics and the talk in the commons. PMID- 11868416 TI - Bioethics and healthcare reform: a Whig response to weak consensus. PMID- 11868418 TI - CQ sources/bibliography. Consensus in bioethics: negotiating the challenge of moral pluralism. PMID- 11868419 TI - Response to "Cutting bodies to harvest organs" by John Portmann (CQ Vol 8, No 3). Autonomy as scapegoat in the organ shortage debate: a reply to Portmann. PMID- 11868420 TI - Consensus formation: the creation of an ideology. PMID- 11868422 TI - Response to "May a woman clone herself" by Jean Chambers (CQ Vol 10, No 2). Are there limits to the use of reproductive cloning? PMID- 11868421 TI - Response to "May a woman clone herself?" by Jean E. Chambers (CQ Vol 10, No 2) and "Entitlement to cloning" by Timothy F. Murphy (CQ Vol 8, No 3). Clone alone. PMID- 11868423 TI - Successes and failures of hospital ethics committees: a national survey of ethics committee chairs. PMID- 11868424 TI - Patient productivity as a value and a variable in geriatric healthcare allocation. PMID- 11868425 TI - Chris Shaw on ethical issues in biotechnology. Interview by Thomasine Kushner. PMID- 11868426 TI - GERD management. Strategies recommended for primary care practice. AB - Primary care physicians are on the front lines in the treatment of gastroesophageal reflux disease. Therefore, they need the most current information available regarding how to achieve the best outcomes possible. Much clinical evidence indicates that when prescribed for initial and maintenance therapy, the proton pump inhibitors are effective for symptom relief and cost effective in maintaining remission. In this article, Drs Kuritzky and Rodney discuss several step management strategies that could well be "just what the doctor ordered." PMID- 11868427 TI - GERD and its complications. The pathogenic relationship between symptoms and disease progression. AB - Chronic daytime and noctural acid reflux causes mucosal damage, heartburn, and other symptoms of GERD. The esophageal complications of GERD result from long term exposure of the esophagus to acid. Extraesophageal GERD complications, such as laryngitis and asthma, develop when some of the acidic refluxate in the proximal esophagus enters the lower or upper airways because of comprised defense mechanisms. A growing body of evidence suggests that nocturnal reflux is more important to the development of severe complications than is daytime reflux. Defects in the lower esophageal sphincter and clearance mechanisms are related in part to recumbency during sleep; they may explain why transient episodes of nocturnal reflux outnumber daytime episodes in patients with GERD. Currently, the hope is that management strategies aimed at control of nocturnal symptoms can prevent, reduce the severity of, or resolve the long-term complications of chronic GERD. PMID- 11868428 TI - Proton pump inhibition. An effective, safe approach to GERD management. AB - Prescribed worldwide to treat gastroesophageal reflux disease (GERD), the proton pump inhibitors (PPIs) not only relieve acid reflux-related symptoms more rapidly than standard-dose or high-dose histamine2 receptor antagonists but also accelerate the rate of esophageal healing. Five PPIs--omeprazole, esomeprazole, lansoprazole, rabeprazole, and pantoprazole--are currently labeled by the Food and Drug Administration. These agents share a common mechanism of action and rarely exhibit clinically important interactions with other hepatically metabolized medications or pH-dependent drugs. Except for lingering concern about their long-term use in Helicobacter pylori-positive patients, the PPIs produce relatively few adverse effects when administered for the short or long term. Because primary care physicians are generally the first to treat patients with GERD, they may find it helpful to expand their knowledge of the pharmacologic effects of the PPIs. With an eye toward this end, Dr Berardi presents a cogent overview of PPI pharmacodynamics, pharmacokinetics, efficacy, drug interactions, and safety. PMID- 11868429 TI - Introduction. GERD warrants increased physician appreciation and improved treatment. AB - Many primary care physicians and the public have long considered GERD to be a nuisance disorder that has a modest negative impact and few long-term complications. Until just recently, both groups also underappreciated the significance of nocturnal heartburn. However, the information presented in this Special Report shows that nocturnal heartburn has a major adverse effect on patients' lives. In fact, most experts in the field of gastroenterology believe that prolonged exposure of the esophagus to the acidic contents of the stomach eventually leads to Barrett's esophagus in certain patients. Over time, Barrett's metaplasia may become dysplastic and then malignant. In addition to the increased risk of these long-term complications, the significant detrimental effects of heartburn on patients' daily activities and quality of life point to the need for more aggressive treatment with more powerful acid suppressants. The efficacy of PPI therapy is dependent on H+/K+ ATPase proton pump kinetics and the AUC of the PPI prescribed. Therefore, administration of a PPI before the first meal of the day is critical, as is careful selection of an agent that will control nocturnal heartburn. PMID- 11868431 TI - [Priorities in resource usage in risk assessment]. PMID- 11868430 TI - [Food and health: general review]. PMID- 11868432 TI - [About implementation of methodology in risk assessment in Russia]. PMID- 11868433 TI - [Safety control of foodstuffs in Belorussia]. PMID- 11868434 TI - [Government system of quality control and safety of food products and food resources in the Ukraine]. PMID- 11868435 TI - [International aspects and harmonization--actuality of risk assessment in international trade]. PMID- 11868436 TI - [Hazard identification]. PMID- 11868437 TI - [Determination of "dose-response'' characterization]. PMID- 11868438 TI - [Risk characterization]. PMID- 11868439 TI - [Specific issues in microbiological risk assessment]. PMID- 11868440 TI - [Safety of foodstuffs: use of risk assessment. Summary report]. PMID- 11868441 TI - A comparison of the quality of Cochrane reviews and systematic reviews published in paper-based journals. AB - This study set out to compare Cochrane reviews and reviews published in paper based journals. Two assessment tools were used to collect the data, a 23-item checklist developed by Sacks and a nine-item scale developed by Oxman. Cochrane reviews were found to be better at reporting some items and paper-based review at reporting others. The overall quality was found to be low. This represents a serious situation because clinicians, health policy makers, and consumers are often told that systematic reviews represent "the best available evidence." In the period since this study, the Cochrane Collaboration has taken steps to improve the quality of its reviews through, for example, more thorough prepublication refereeing, developments in the training and support offered to reviewers, and improvements in the system for postpublication peer review. In addition, the use of evidence-based criteria (i.e., the QUOROM statement) for reporting systematic reviews may help further to improve their quality. PMID- 11868442 TI - A brief history of research synthesis. AB - Science is supposed to be cumulative, but scientists only rarely cumulate evidence scientifically. This means that users of research evidence have to cope with a plethora of reports of individual studies with no systematic attempt made to present new results in the context of similar studies. Although the need to synthesize research evidence has been recognized for well over two centuries, explicit methods for this form of research were not developed until the 20th century. The development of methods to reduce statistical imprecision using quantitative synthesis (meta-analysis) preceded the development of methods to reduce biases, the latter only beginning to receive proper attention during the last quarter of the 20th century. In this article, the authors identify some of the trends and highlights in this history, to which researchers in the physical, natural, and social sciences have all contributed, and speculate briefly about the "future history" of research synthesis. PMID- 11868443 TI - Egg on their faces. The story of human albumin solution. AB - In 1998, the Cochrane Injuries Group published the results of a systematic review of human albumin administration in critically ill patients. The results showed that the risk of death in patients receiving albumin was 14%, and the risk of death in patients not receiving albumin was 8%, suggesting that for every 17 critically ill patients treated with albumin there is one extra death. The results were widely reported in the television and print media throughout the world and stimulated an immediate response from the drug regulatory agencies, the plasma products industry, and the medical profession. Despite vigorous attempts by the plasma products industry to limit the effect of the systematic review on albumin sales, the use of albumin declined steeply, showing that evidence from systematic reviews can have an important effect on clinical care. PMID- 11868444 TI - Development of the Cochrane Collaboration's CENTRAL Register of controlled clinical trials. AB - The Cochrane Collaboration has established a centralized database of controlled trials and other studies of health care interventions (called CENTRAL) that serves as the best available resource for all those preparing and maintaining systematic reviews or otherwise searching for trials. CENTRAL is available on The Cochrane Library. This article describes the history and methods of CENTRAL's development and the results of an analysis of the current composition of CENTRAL. As of September 2000, CENTRAL contained almost 300,000 citations to reports of trials, contributed mainly by Cochrane Groups and Centers around the world. Development of CENTRAL has been an ambitious, scholarly undertaking and has resulted in a valuable resource: CENTRAL includes citations to controlled trials that may not be indexed in MEDLINE, EMBASE, or other bibliographic databases; citations published internationally in many languages; and citations that are available only in conference proceedings or other hard-to-access sources. PMID- 11868446 TI - The contribution of handsearching European general health care journals to the Cochrane Controlled Trials Register. AB - A fundamental aim of any systematic review is that all relevant studies should be identified and considered for inclusion. Limitations with searching bibliographic databases led the Cochrane Collaboration to search journals by hand for reports of trials. This article presents the results of a 3-year project to identify and make accessible reports of randomized trials published in European general health care journals. Overall, 21,620 reports of controlled trials were identified from 119 journals from 16 countries. More than three quarters (76%) were published in U.K. or German journals. Only 3,640 (17%) reports were indexed in MEDLINE as controlled trials, and 6,554 (30%) were not indexed in MEDLINE at all. Bibliographic details for all reports are available by searching The Cochrane Controlled Trials Register in The Cochrane Library. This project has ensured that a large proportion of trial reports not previously identifiable has been made accessible to those preparing systematic reviews. PMID- 11868447 TI - To IPD or not to IPD? Advantages and disadvantages of systematic reviews using individual patient data. AB - Systematic reviews and meta-analyses that obtain original research data on individual participants enrolled in trials have been described as the gold standard of review. However, they may take longer and be more resource intensive than other types of review. The authors describe potential advantages and disadvantages of the individual patient data (IPD) approach, including benefits from improved data quality, benefits afforded by the type of analyses that can be done, and advantages in achieving consensus around results and interpretation by an international multidisciplinary team. Disadvantages and barriers relating to resource and expertise, negotiating collaboration, and software requirements are also discussed. At the outset, reviewers should consider the methodological factors likely to influence results in their particular review setting, together with time and resource constraints, so that an active decision can be made about whether to extract data from published reports, collect additional or replacement summary data from trialists, or collect IPD. PMID- 11868448 TI - The Cochrane Collaboration: providing and obtaining the best evidence about the effects of health care. PMID- 11868449 TI - The Criticism Management System for the Cochrane Library. AB - In April 1997, the Cochrane Collaboration implemented the Criticism Management System for The Cochrane Library, allowing readers to electronically submit criticisms on Cochrane reviews and protocols. As a result of resource constraints and user feedback, the Criticism Management System evolved through three different versions. Major problems have included difficult access to the system, a lack of automation within the system, a low number of submissions (171 criticisms submitted per 1,388 reviews), and a low response rate to these criticisms by authors of reviews (26 criticisms have received published responses). These low figures suggest that users of The Cochrane Library are not commenting on the published reviews or not using the Criticism Management System to make their criticisms known to the authors. An easier-to-use system and better ways of encouraging feedback from readers and responses from reviewers are needed in the future. PMID- 11868450 TI - [Clinical proarrhythmias induced by anti-arrhythmic drugs, non-cardiovascular agents and implantable cardioverter-defibrillators]. AB - The authors survey the electrophysiological mechanisms and clinical spectrum of the aggravation of cardiac arrhythmias provoked by antiarrhythmic drugs, non cardiovascular agents and implantable cardioverter-defibrillators. The main aspects of the recognition, the differential diagnostics and the therapy are discussed. It is demonstrated that prevention of drug-induced arrhythmogenesis necessitates a fundamental understanding of the risk factors of proarrhythmias and of the clinical pharmacological properties of the drugs that are intended to be used, including proarrhythmic drug interactions. A systematic account is presented of those drugs prescribed in non-arrhythmic diseases which may lead to out-of-hospital cardiac arrest. The proarrhythmias that may be observed in recipients of implantable cardioverter-defibrillators are discussed, as are the undesirable interactions between these devices and antiarrhythmic drugs in such patients. PMID- 11868452 TI - [Surgical treatment of non-palpable breast tumors]. AB - METHOD: Between 1997 and 2000 the authors performed 110 fine-wire localized breast biopsies in patients with nonpalpable suspicious lesions. RESULTS: 108 target lesions (98.1%) were accurately excised during the initial surgery. Overall, 56 lesions were malignant, among these cases 50 invasive carcinomas and 6 DCIS were found. Breast conservation was achieved in 48 patients (88.5%), simple mastectomy with axillary lymph node sampling was performed in 8 cases. 86% of the invasive malignant lesions belonged to the good prognostic subgroup of the Nottingham Prognostic Index. CONCLUSION: There in an importance of the close cooperation between radiologist, surgeon, pathologist and oncologist in the treatment of nonpalpable preclinical breast carcinomas. PMID- 11868451 TI - [Acute toxicological cases during a ten-year period in our clinic]. AB - INTRODUCTION: There's a fact, that Hungary has held the first places in suicidal statstics. METHODS: The authors studied toxicological cases between 1989 and 1998 at the 1st Department of Medicine of the Medical and Health Science Centre, at the University of Debrecen, paying special attention to suicidal poisoning cases. RESULTS: 2% of the patient turnover accounted for acute poisoning cases, the number of which increased during the 10 years in question. 70% of the cases were of suicidal intentions, 20% were unintentional, these poisonings were not committed on purpose, while the proportion of iatrogenic intoxication cases was 10%. Amongst the failed suicide cases there was a higher proportion of women, whereas a higher percentage of men accounted for "successful" suicide cases. When examining auto-intoxication cases it turned out that the medicine most frequently used was meprobamate, besides benzodiazepines. Mortality rate was highest in the glutethimide intoxication cases. Most poisonings with suicidal intentions took place in the 2nd quarter of the year. Most completed suicides were committed on Wednesdays and Thursdays. 81% of the iatrogenic intoxication cases happened to be with digitalis and coumarin overdose. Nearly 50% of the cases turned out to be combined intoxications. 40% of the men took alcoholic drinks during the auto intoxications. In the case of 135 patients extracorporeal detoxification therapy was applied, which consisted mostly of hemoperfusion. Three quarters of the patients needed psychiatric care and every fourth patient was admitted to the Department of Psychiatry. 6.9% of the poisonings were fatal. CONCLUSIONS: The growing number of toxicological cases--amongst these suicidal poisonings--compels us to pay more attention to the setting up of interdisciplinary based prevention as well as running effective toxicological centres. All physicians have a responsibility to recommend psychiatric care for people suffering from mental problems or depression and for the unsuccessful or potential suicide seeking help for the first time. Family doctors in primary medical care and who meet patients first have an important role in this job. PMID- 11868453 TI - [Hepatitis viruses and hepatocarcinogenesis]. AB - INTRODUCTION: Hepatocellular carcinoma (HCC) is one of the most frequent malignancies worldwide. Hepatitis viruses, as the hepatitis virus B (HBV) and hepatitis virus C (HCV), among other environmental factors, are undoubtedly listed in the etiology of HCC. Studies indicate that in the near future viral hepatitis will be of increasing importance in the etiology of HCC. AIM: This review briefly discusses the known carcinogenic effects of HBV and HCV in the light of experimental and human studies. RESULTS: Data show that viral proteins may directly interfere with gene products responsible for cell proliferation and cell growth, therefore this direct effect may influence the basic mechanism of the cell cycle. Many other signal transduction cascades may be affected as well. Direct integration of HBV viral sequences into the host genome increases the genomic instability. The genomic imbalance allows the development and survival of malignant clones bearing defected genomic information. CONCLUSION: HBV and HCV infection induce indirect and direct mechanisms through cellular damage, increased regeneration and cell proliferation, therefore enhancing the development of HCC. PMID- 11868454 TI - [Severe congenital factor V deficiency: case report]. AB - INTRODUCTION: Congential deficiency of factor V is a rare condition, transmitted in autosomal recessive way. Heterozygote patients generally have no symptoms, homozygotes present with spontaneous and postoperative bleedings. About one-half of patients are diagnosed in adulthood. METHODS: The presented case is a double heterozygote, which was confirmed using moleculare biological methods. The authors found many carriers in his family. CONCLUSIONS: This rare and sometimes severe disorder needs regular haematological controls. Carriers must have more attention during surgical procedures and labour, and the family doctor should be informed about there condition. PMID- 11868455 TI - [History of the Physiology Department of the Budapest University]. PMID- 11868456 TI - [The effect of image analyser noises in studies of cell structure]. AB - Using image analysers, the influence of noises on the quality of images obtained from three types of digital CCD videocameras was studied. Algorithms for calculating the heterogeneity coefficient of cell structures have been proposed, which take into account the noises on the images. Application of procedures of image smoothing or averaging from a few shots, calculation of differences in heterogeneity coefficients of the object and a free field, and a combination of these methods have significantly reduced the influence of noises and increased the informativeness of texture features. PMID- 11868458 TI - [Structural characteristics of nematocyst stinging threads from the parasitic cnidarian Polypodium hydriforme]. AB - The structure of discharged nematocyst stinging threads present in free-living individuals of Polypodium hydriforme was studied by scanning and transmission electron microscopy. Not all cnidae of P. hydriforme proved to be atrichous isorhizas (as previously was accepted), but only one of the five nematocyst categories studied. A unique feature of P. hydriforme nematocysts was revealed: their stinging threads possess two strands of spines, rather than 5 or 3, as in Narcomedusae and other Cnidarians, respectively. This fact supplements the evidence in favour of P. hydriforme being a rather isolated branche in the phylogenetic tree of Cnidaria. PMID- 11868457 TI - [Ultrastructure and morphology of the mitochondriome of cardiomyocytes from invertebrates. I. The mitochondriome of cardiomyocytes from insects]. AB - The cardiomyocyte mitochondrial ultrastructure of two insect species (the American cockroach Periplaneta americana, and a dragonfly Aeschna sp.) has been studied. Mitochondria in cardiomyocytes of these insects are connected by intermitochondrial contacts, similar in morphology to vertebrate intermitochondrial contacts. The number of intermitochondrial contacts differs in cardiomyocytes of the studied insects, numbering 12 and 18 per 100 mitochondria in cardiomyocytes of the cockroach and dragonfly, respectively, which is due presumably to differences in activity of these insects. Cardiomyocytes of both species have several features in common. It was shown that cross-striated myofibrils oriented in different directions occupy 50-58% of the cytoplasmic volume, while mitochondria cover only 16-18%. The pattern of mitochondrial localization differs in cardiomyocytes of the two studied insects. In the cockroach, cardiomyocyte mitochondria are seen both in the center of the cell and on its periphery, in protrusions; whereas in the dragonfly, mitochondria of cardiomyocytes are confined to the protrusions of the abluminal cell side. Mitochondrial profiles are small, their packing is not dense. Mitochondria in cardiomyocytes of these insects have few plastic cristae and dense matrix. PMID- 11868459 TI - [Determination of a position of a functional pore in the tobacco pollen]. AB - Pollen hydration and germination on the "wet" stigma of Nicotiana tabacum L. were studied by SEM and TEM to reveal the role of the stigma in selecting the germinative pore, and in establishing the axis of polarity in the pollen grain. Pollinated stigmas were fixed with glutaraldehyde or osmium tetroxide vapour, or processed with rapid freeze fixation and freeze substitution. Fixation was performed in 5, 15 or 30 min and 3.5 h after pollination. The tube easily emerged from either pore, this process not depending on the pollen grain orientation relative to the stigma. The orientation of pollen tubes remained random till their length becomes longer than the pollen grain diameter. The TEM analysis of ultrastructural changes in poral regions during pollen hydration and germination showed that the germinative pore was positioned just near the generative cell and vegetative nucleus. Within the first 5 min after pollination a new layer of the electron-lucent wall adjacent to the plasma membrane was formed in the region of a future germinative pore. Following 15 min, marked changes were revealed in the cytoplasm region, close to the germinative pore. Minute dictyosome vesicles were accumulated near the plasma membrane. Small mitochondria and short ER cisternae were distal to a zone of secretory vesicles. The data suggest that the axis of polarity in the germinating pollen grain is predetermined by a spatial organization of the vegetative cell. PMID- 11868460 TI - [Penetration of lipoproteins through a capillary wall and their effects on the cardiomyocyte ultrastructure during in vitro heart perfusion]. AB - The colloidal gold-labeled lipoprotein (LP) distribution in the myocardium after the 30 min perfusion of isolated working rat heart was studied by electron microscopy. LP of physiological concentrations are shown to be able to interact with the capillary wall and they can be incorporated by endotheliocytes. The velocity and transportation mechanisms of different classes of LP differ. Very low density LP are the most intensively taken up, low density LP enter into interstitium more rapidly than others, high density LP do not leave the capillary wall at all. The passages of labeled LP through endotheliocytes by the receptor mediated as well as the non-receptor manners are revealed. The transportation through the widened intercellular junctions may be supposed for low density LP. For the fist time LP addendum into the perfusion medium was shown to provoke the activation of interstitial macrophages. During the perfusion duration they take inside and accumulate labeled very low density LP and low density LP in their lysosomes. The internalization may be performed by the specific endocytosis or by the simple phagocytosis. The qualitative and morphometrical analyses show that LP preserve the capillary and cardiomyocyte ultrastructure from perfusion injuries. One may suppose that there are interrelations between the capillary endothelium, the interstitial macrophages and the parenchymal cells in myocardium for realization of plasma LP effects. PMID- 11868461 TI - [Effect of actomyosin contractility on focal contacts of myofibroblasts and structure of stress fibers]. AB - Myofibroblasts from rat lung were cultivated. These cells in addition to beta- and gamma-cytoplasmic actins, expressed alpha-smooth muscle actin (alpha-SMA) and formed a system of "supermature" focal contacts, which were connected with thick stress-fibers expressing alpha-SMA and myosin II. Reduction of actin-myison contractility by inhibitors BDM and ML-7 lead to stress fiber reorganization, e.g., decrease in their thickness, a selective disappearance of alpha-SMA expression and myosin translocation from bundles to the cytoplasm. Using immunofluorescence, interference-reflection microscopy and morphometry, we have demonstrated that an inhibition of actin-myosin contractility also leads to dispersion of myofibroblastic focal contacts. Phase-contrast and DIC video enhanced microscopy of live cells showed morphological reorganization at the leading edge after inhibitory treatment. Thus, actin-myosin contractility controls the structure of "supermature" focal contacts of myofibroblasts and alpha-SMA expression in stress fibers. PMID- 11868462 TI - [Characteristics of nucleolar organization and function under condition of complete spatial in situ disassembly of its main structural components]. AB - It is well known that fibrillar centers (FC) constitute an essential structural component of the active nucleolus in mammalian cells, yet their role in regulation of ribosomal gene transcription still remains an open question. Here, we studied the activity of endogenous RNA polymerase I upon partial and complete unraveling of nucleoli and FCs. The pattern of BrUTP incorporation in nuclei of hypotonically-treated cells was shown to be essentially the same as in the control untreated cells. Moreover, the sites of BrUTP incorporation, which revealed the active PNA polymerase I, were completely coincident with UBF-binding sites. These observations allow to conclude that structural integrity of FCs is not a prerequisite for maintenance of the active RNA polymerase I transcriptional complex. When the action of hypotonic shock was ceased and the cells were transferred to a complete cultural medium, the swollen nucleoli recovered to the control state. Therefore it is possible to conclude that none of the main morphological nucleolar counterparts, such as FCs, dense fibrillar component or the pars granulosa, is responsible for the maintenance of the nucleolar structural and functional integrity. A suggestion is made that this role may be played by the nucleolar matrix associated with the RNA polymerase I transcriptional complex. PMID- 11868464 TI - [The reaction of acclimated isolated gill epithelium of mollusc to a short-term superoptimal heating]. AB - Thermoresistance (TR) of isolated gill epithelium of molluscs Anodonta anatina L. (n = 20) acclimated for 72 h at 24 degrees C was studied. In 1, 2, 12, 24, 36, 48, 60, and 72 h, the gills were submitted to a provoking action: a 10-minutes heating at 36 degrees C. As a criterion of the tissue TR level served a logarithmic index of the time of survival at 40 degrees C. During 24 h, negative correlation (about 0.50) was revealed between the acclimated epithelium TR level and the value and direction (sign) of its change under the provoking action. In 36 h, this coefficient fell to 0.13. As a result, the range of interorganism variability of the tissue TR level enlarged. Meanwhile, a decrease in the acclimated tissue mean TR level occurred as late as in 60 h of the experiment, being equal to 21%. In 72 h, it reached 30%, which traditionally indicates a deterioration of the functional state of tissue cells and a subsequent death. However, we believe that the deterioration of this state actually had happened earlier, in 36 h, when the range of the interorganism variability of the tissue TR level became enlarged. It was at this moment that the acclimated tissue lost its ability to regulate the shift in stability with the TR level at this period of acclimation. PMID- 11868465 TI - [Atypical transformation of cell populations after low irradiation and other actions]. AB - Ionizing irradiation at low doses and some weak non-irradiational (including non mutagenic) actions can produce a peculiar cell reaction, such as a massive, dose independent transition to a new regime of existence. The changes appear saltationally. They are maintained for a very long time by both rarely and actively dividing cells. The most characteristic phenotype of the studied transformation is a stable increase of probability of cell damage and death. Some other manifestations of this transformation have been also revealed. This unusual response is found upon observation of totally different biological objects (unicellular organisms, cells of various mammalian tissues) to be regarded as a general biological regularity. The role of such radiational alterations in pathogenesis of remote consequences of low-dose irradiation is discussed. PMID- 11868463 TI - [Adenylyl cyclase pathway is involved in the regulation of intracellular calcium level regulation in brown preadipocytes]. AB - Mechanism of adrenergically activated calcium response in freshly isolated brown preadipocytes was studied with fluorescent probe Fura-2. Application of a direct activator of adenylylcyclase forskolin or cell permeable analog BrcAMP caused rise in the intracellular calcium level that was even higher than after the application of norepinephrine. Protein kinase A inhibitor H-89 in a dose dependent manner reduced, while inhibitor of total phosphodiesterase activity IBMX, or protein phosphatase inhibitor ocadaic acid enhanced norepinephrine or isoproterenol initiated cellular calcium responses. It is concluded that cAMP and protein kinase A mediated phosphorylation play a crucial role in adrenergically initiated calcium signalling in brown preadipocytes. PMID- 11868466 TI - [Chromosomal abnormalities in lymphocytes from a patient with Werner's syndrome in intact culture and after treatment with halogenated analogs of thymidine]. AB - Four balanced chromosomal translocation, deletion of chromosome 15, and a break in chromosome 11 were detected in 100 G-banded metaphases of cultured lymphocytes of a patient with Werner's syndrome. We observed aneuploidy that included both trisomies and monosomies for various chromosomes. Halogenated analogs of thymidine in low doses increased significantly the incidence of chromosome aberrations accompanied by fragments. 5-Iododeoxyuridine induced lesions in centromeric regions of B-group chromosomes in 44.4% of all the cases of breaks. A hypothesis is proposed about the existence of a special mechanism for genetic control in changes in the cell nucleus and mitotic chromosome transformation. This mechanism can be manifested after the application of halogenated analogs of thymidine. The mutation involved in Werner's syndrome is presumably related to this mode of genetic control. PMID- 11868467 TI - [The functional morphology of polytene chromosomes in the midge Chironomus pilicornis F. from the permafrost zone reservoirs]. AB - The karyotype and chromosomal polymorphism of Chironomus pilicornis from the reservoirs of Yakutian permafrost zone are described. In the Yakutian populations, of 10 inversion sequences 11 genotypical combinations of these were registered. The level of inversion heterozygosity is 50-70%, but in several populations it makes only 11-29%. In comparison with the Scandinavian populations, significant functional modifications of chromosomal morphology were found. They are associated with the increase in centromeric heterochromatin amount, facultative chromocentre formation, and the appearance of numerous B chromosomes and nucleoli, including the facultative ones. A possible adaptive significance of such functional modifications conditioned by climate peculiarities is discussed. PMID- 11868468 TI - [Evidence of species specific pericentric inversion in the karyotype of the midge Chironomus balatonicus]. AB - Pericentric inversions do not play any important role in chromosomal rearrangements in the karyotype evolution of the genus Chironomus. However, a unique case of the fixed pericentric inversion was discovered in chromosome 2 of Chironomus balatonicus--one of the members of plumosus-species group (Kikhadze et al., 1996a; Golygina et al., 1996). According to morphological criteria, a centromere band on chromosome 2 changed its position in Ch. balatonicus. The cloned H3-SauDNA, specific for centromeres in plumosus group, was in situ hybridized with Ch. balatonicus polytene chromosomes, and thus a real change in the centromere position was proved to be a result of pericentric inversion. This was also confirmed after differential C-staining. PMID- 11868469 TI - Tuberculosis of the maxillary sinus manifesting as a facial abscess. AB - Tuberculosis of the maxillary sinus is rare. Likewise, an acute onset that necessitates incision and drainage is also very uncommon. We report the case of a 15-year-old girl who came to us with an abscess on the left side of her face. She was found to have tuberculosis of the left maxillary antrum. PMID- 11868470 TI - Seborrheic keratosis of the auricle: report of an unusual case. AB - Seborrheic keratoses are superficial epithelial lesions. They are usually pigmented and often warty, but sometimes they appear as smooth papules. These lesions frequently occur on the face during middle and old age. We report an unusual case of a large seborrheic keratosis of the auricle. We also emphasize the importance of the differential diagnosis, and we discuss the treatment strategy for these usually benign but disfiguring tumors. PMID- 11868471 TI - Extramedullary plasmacytoma manifesting as a palpable mass in the nasal cavity. AB - We report a rare case of plasmacytoma of the nasal cavity in a 60-year-old man. The patient had a history of a steadily growing and palpable mass in the opening of the left nostril. The tumor was found to be localized, and it was identified as an extramedullary plasmacytoma. The patient underwent therapy with 55 Gy of radiation. Six months following the cessation of radiotherapy, the size of the tumor had not changed. The mass was then completely excised under microscopic vision. Histopathologic examination identified the growth as a plasmacytoma with monotypic light-chain expression. To optimize the management of patients with an extramedullary plasmacytoma of the head and neck, interdisciplinary management is mandatory. PMID- 11868472 TI - Target-specific multimodality endovascular management of carotid artery blow-out syndrome. AB - We describe a novel multimodality endovascular approach to safely control hemorrhage from a carotid artery pseudoaneurysm and tumor vasculature associated with a squamous cell carcinoma. This approach was used in the case of a 68-year old man who had previously undergone a laryngectomy, chemotherapy, and brachytherapy and who subsequently experienced acute oropharyngeal bleeding. Angiography detected a carotid artery pseudoaneurysm and significant tumor vascularity. A target-specific multimodality approach was taken to embolize the potential etiologies for both the current and any future hemorrhages. Stent assisted coiling of the pseudoaneurysm was successful. The tumor blush was treated with polyvinyl alcohol particles and both retrievable and nonretrievable coils. Endovascular surgeons have become increasingly involved in the management of patients with carotid injuries and with neoplasms in and around the skull base. Current endovascular technology provides a rapid target-specific approach to the treatment of carotid artery blow-out syndrome and has a greater potential to lower morbidity than does carotid sacrifice. PMID- 11868473 TI - The fragility of life. PMID- 11868474 TI - Delayed reocclusion of a middle meatal antrostomy. PMID- 11868475 TI - A posthemorrhagic mass, a contralateral polypoid mass, and unilateral laryngeal nerve paresis. PMID- 11868476 TI - Chondrosarcoma of the larynx. PMID- 11868477 TI - Caloric test results in a patient with Meniere's syndrome. PMID- 11868478 TI - Dilation of an esophageal stricture caused by epidermolysis bullosa. PMID- 11868479 TI - Clean claims are key to timely reimbursement, but be vigilant. PMID- 11868480 TI - Endotracheal tube fire during tracheostomy: a case report. AB - Electrosurgery in the presence of volatile anesthetic gases has been associated with operating-room fires. We report a case in which an operating-room fire occurred while an intubated patient underwent electrosurgical tracheostomy. The fire in this case was caused by a combination of an oxygen-rich environment, a polyvinyl chloride tube, and heat generated by an electrosurgical unit. We also discuss factors that increase the risk of this type of fire and the management steps that should be undertaken in the event that such a fire occurs, and we briefly review the literature on this subject. PMID- 11868481 TI - Nasolabial cyst: a retrospective analysis of 18 cases. AB - Nasolabial cysts are rare but easily identifiable when they do occur. They are thought to arise from the remnants of the nasolacrimal ducts, but most of the available information on these cysts is limited to isolated case reports. The purpose of our study was to examine the clinical and pathologic features of nasolabial cysts in order to provide a basis for their correct diagnosis and treatment. Eighteen patients with nasolabial cysts were treated at Korea University's Guro Hospital between Jan. 1, 1988, and Dec. 31, 1999. We retrospectively analyzed the clinical appearance, pathologic findings, and treatment results in each case. All patients underwent surgery via the sublabial approach. The lining epithelia were identified as either pseudostratified, stratified squamous, or simple cuboidal. A cyst recurred in only one case, and there were no surgical complications on follow-up. The results of our study provide a basis for sound diagnosis and treatment. PMID- 11868482 TI - Local injection of OK-432 in the treatment of ranula: a case report. AB - We treated a 57-year-old woman for ranula. After aspirating the cyst's contents, we administered 0.1 KE/ml of OK-432 via local injection. One month later, the cyst had still persisted, so we repeated the procedure. After 2 weeks, the ranula began to shrink markedly, and at 4 weeks it had almost disappeared. No recurrence of the ranula was observed during the subsequent 1 year of follow-up. Following each injection, the patient developed transient fever and local swelling but no serious complications. Our experience suggests that OK-432 injection is an effective treatment for ranula. However, because this treatment causes the cyst to collapse rather than disappear completely, patients should be regularly monitored over the long term. PMID- 11868483 TI - Facial palsy from metastatic nasopharyngeal carcinoma at various sites: three reports. AB - Nasopharyngeal carcinoma that causes clinically evident facial palsy is uncommon. This article describes and discusses a series of cases that illustrates how nasopharyngeal carcinoma caused facial palsy as a result of facial nerve involvement at three sites: the cerebellopontine angle, the middle ear, and the parotid. The maxim, "All that palsies is not Bell's," is particularly relevant with respect to patients who have previously been treated for advanced nasopharyngeal carcinoma. In these patients, recurrent or persistent nasopharyngeal carcinoma involving the cerebellopontine angle, temporal bone, or parotid should be excluded. PMID- 11868484 TI - The legacy of Chinese herbal medicine. PMID- 11868485 TI - Traditional Chinese medicine in contemporary China. PMID- 11868486 TI - The doctor's garden. PMID- 11868487 TI - Medical acupuncture in the twenty-first century. PMID- 11868488 TI - Homeopathy: the other medicine. PMID- 11868490 TI - An alternative view of complementary and alternative medicine. PMID- 11868491 TI - Magnets and elixirs. PMID- 11868492 TI - Reading clinical research--what can we expect to learn from reading randomized clinical trials? PMID- 11868493 TI - Employee training and job descriptions. PMID- 11868494 TI - Ensuring the health of Maryland's citizens: the 2002 legislative agenda. PMID- 11868495 TI - A history of the Baltimore County Medical Association. PMID- 11868499 TI - Categories of complementary and alternative medicine. PMID- 11868500 TI - Iatrogenic thyrotoxicosis. Causal circumstances, pathophysiology and principles of treatment. Review of the literature. AB - Thyrotoxicosis is the clinical syndrome that results when tissues are exposed to high levels of circulating thyroid hormones. In most instances, thyrotoxicosis is due to hyperthyroidism, a term reserved for disorders characterized by overproduction of thyroid hormones by the thyroid gland. Nevertheless, thyrotoxicosis may also result from a variety of conditions other than thyroid hyperfunction. The present report focuses on the etiologies, pathophysiology and treatment of iatrogenic thyrotoxicosis. Iatrogenic thyrotoxicosis may be caused by 1) subacute thyroiditis (a result of lymphocytic infiltration, cellular injury, trauma or radiation) with release of preformed hormones into circulation, 2) excessive ingestion of thyroid hormones ("thyrotoxicosis factitia"), 3) iodine induced hyperthyroidism (radiological contrast agents, topical antiseptics or other medications). Among these causes of iatrogenic thyrotoxicosis, that induced by the iodine overload and cytotoxicity associated with amiodarone represents a significant challenge. Successful management of amiodarone-induced thyrotoxicosis requires close cooperation between endocrinologists and endocrine surgeons. Surgical treatment may have a leading yet often underestimated role in view of the potential life-threatening severity of this disease, whereas others kinds of iatrogenic thyrotoxicosis are usually treated conservatively. PMID- 11868501 TI - Long-term results of surgical resection of lung metastases. AB - Between 1990 and 2000, 56 consecutive patients underwent lung resection for removal of metastatic disease. Mortality, disease-free interval, and overall survival were studied. Only patients with a complete follow-up were included and data were collected conform the protocol of the International Registry of Lung Metastases. The primary tumour in our series was an epithelial tumour in 25 patients (45%), sarcoma in 15 (27%), germ cell tumours in 11 (19%) and melanoma in 5. Operative mortality was 1.4% (1 out of 73 procedures). Germ cell tumours had the best survival (76% at 5 years), and melanoma the worst (0% at 5 years). Multivariate analysis showed that survival for patients who underwent 2 or more metastasectomies was surprisingly good with a 5-year survival rate of 46%. Survival was not related to disease-free interval, multiple lung metastases, or pneumonectomy. It is in accordance with some reports that a short disease-free interval, numerous lung metastases, or recurrence after the first metastasectomy should not preclude patients from operation. PMID- 11868502 TI - Current outcome in patients with lung cancer and positive mediastinoscopy. AB - PURPOSE: A retrospective study was undertaken to determine the contemporary treatment and outcome of patients with lung cancer and positive lymph nodes proven at mediastinoscopy. MATERIAL AND METHODS: From 1994 to 1999 a cervical mediastinoscopy was performed in 361 patients. Metastatic lymph nodes from a bronchogenic carcinoma were present in 81 patients; 71 were men. Mean age was 61.7 years. Squamous cell carcinoma and adenocarcinoma were present in 26 patients each (32.1%), large cell carcinoma in 19 patients (23.4%), small cell carcinoma in six patients (7.4%) and other histologies in four patients (5%). Stage IIIA was found in 51 patients (69.9%), IIIB in 17 (23.3%) and IV in 5 (6.8%). Of eight patients no precise stage could be determined (9.9%). These patients were retrospectively reviewed. Survival time was analyzed according to the Kaplan-Meier method. A multivariate Cox analysis was performed to determine significant factors in relation to survival. RESULTS: Forty-four patients (54.3%) were treated by induction chemotherapy, of which 15 were operated on, 11 patients (13.6%) had chemotherapy only, 11 (13.6%) radiotherapy only, 3 (3.7%) combined chemo- and radiotherapy, 1 patient (1.2%) had an operation only, and 11 (13.6%) received palliative treatment. Follow-up was complete; 16 patients (19.8%) are alive and 65 (80.2%) died, mostly of local recurrence and metastases. Median survival time (MST) for all 81 patients was 12 months [95% confidence interval (CI) 10-14 months.]. MST for the 15 patients treated by induction chemotherapy + surgery was 27 months [95% CI 18-36 months] and for the 15 patients treated by induction chemotherapy + radiotherapy 15 months [95% CI 12-18 months]. The difference between these two subgroups was significant (p = 0.03). For the 11 patients who had palliative treatment MST was 6 months [95% CI 4-8 months]. The other subgroups had a MST of 10 months or less. In a multivariate Cox analysis only specific treatment (p = 0.0002) and stage (p = 0.02) were found to be significant. CONCLUSIONS: Outcome of patients with lung cancer and positive mediastinoscopy remains poor. In this retrospective study best results were obtained by induction chemotherapy + surgery. An effort should be made to include as many of these patients as possible in neoadjuvant trials. PMID- 11868503 TI - Thoracic trauma: an analysis of 187 patients. AB - A retrospective analysis of 187 cases of thoracic trauma seen between January 1, 1994 and June 30, 1999 is presented. The majority of the patients were male (male female ratio 2.9:1) and the average age at admission was 41.1 years. Blunt trauma, especially motor vehicle accidents (72.2%) and falls (17.1%), were the most frequent causes of chest injury (95.8%). We used the injury severity score (ISS) to assess the severity of trauma. The average ISS for the total group was 27.8 (ranges: 4-75). In only 17.6% of the patients an isolated thoracic trauma was present. Rib fractures (n = 133), pulmonary contusion (n = 110), pneumothorax (n = 78) and haemothorax (n = 65) were the most frequent lesions. Most patients (97.9%) were admitted to the intensive care department. A minority of the patients required thoracotomy (n = 19, 10.2%). Main indications for thoracotomy were pulmonary laceration (n = 5), aortic rupture (n = 3) and rupture of the diaphragm (n = 3). For the majority of cases, observation and/or tube thoracostomy (52.4%) and/or mechanical ventilation (61.0%) were sufficient. Pneumonia and adult respiratory distress syndrome were the most common complications (38.0 and 7.0% respectively). The overall mortality rate was 16.6%. Main causes of death were intracranial hypertension, sepsis combined with multiple organ failure, and hypovolaemic shock. For patients who did not survive the average ISS was 40.3. In a survival analysis the ISS was found to be the most significant determining survival (p < 0.0001), followed by neurotrauma (p = 0.05). Mortality after thoracic trauma remains relatively high, especially in case of associated neurotrauma. The ISS is a valuable score for assessing the severity of trauma and predicting outcome. PMID- 11868504 TI - Hydatid cyst: an unusual disease of the mediastinum. AB - BACKGROUND: Hydatid cystic disease is still a significant clinical problem in endemic countries, particularly in Morocco. Mediastinal localization is very rare. We report our surgical experience with this particular lesion. PATIENTS AND METHODS: In a retrospective review of the last ten years (1990-1999), we had seven cases of mediastinal localization out of total of 2332 (0.3%) intrathoracic hydatid cysts treated in our service. Imaging investigation consisted of chest x ray and computed tomography. All diagnoses were confirmed peroperatively macroscopically and for some lesions, histologically. RESULTS: Complete resection of the cyst was performed in six cases. In one case, a small part of the wall intimately adherent to the superior vena cava was left in place. There were no complications, mortality or recurrence. CONCLUSION: Hydatid cysts can be found in many different sites in the body. Whenever possible, surgical management is the best treatment but extensive resection should be avoided. Medical treatment as an adjunct to surgery is indicated when there is a risk of dissemination. PMID- 11868505 TI - Posterior wall fractures of the acetabulum: characteristics, management, prognosis. AB - A consecutive series of 60 fractures of the posterior wall of the acetabulum, treated operatively in a Level I Trauma center, is reviewed retrospectively. Characteristics of the lesion, type of treatment, early and late postoperative complications and two-year functional results were recorded. In 27 patients (45%), additional damage to the cartilage of the acetabular cavity such as subchondral impaction, free articular fragments or separation of the posterior wall into several pieces was present. Seven patients (11.6%) showed preoperative neurological deficit. All fractures were treated with open reduction and internal fixation through a Kocher-Langenbeck approach. Secondary nerve damage was present in 8.3%. Early secondary surgery was necessary in 8.3%. During the first two years, additional surgery was performed in 7 patients (15.2%). The rate of periarticular ossifications was 26.1%. The rate of excellent and good results was 69.5%. The posterior wall fracture of the acetabulum is a more complex injury than generally is accepted. A large variety of articular lesions with varying degree of complexity is collected in this fracture type. Even in experienced hands, excellent and good long-term results will not exceed 75%. Poor results are due to suboptimal reconstruction of the posterior wall, partial osteonecrosis and/or complications of the Kocher-Langenbeck approach. PMID- 11868506 TI - Laparoscopic cholecystectomy for acute cholecystitis in geriatric patients. AB - Although acute cholecystitis (AC) in many centers is routinely treated by laparoscopic cholecystectomy (LC), the outcome of LC for AC in geriatric patients (75 years or more) remains almost unstudied. All 32 geriatric patients undergoing a cholecystectomy for histologically proven AC in a teaching hospital during a six-year period were studied retrospectively. Median preoperative duration of symptoms was eight days and median preoperative hospital stay was six days. Preoperative ERCP was performed in 22 patients with successful sphincterotomy and common bile duct (CBD) stone retrieval in 11 patients. Overall twelve patients (37%) had CBD stones and 14 patients (44%) had gangrenous cholecystitis at operation. Twenty-seven patients underwent a LC with a conversion rate of 26%, a complication rate of 41% and a mortality rate of 3.7%. Five patients were judged unstable for a laparoscopic approach and underwent a straight open cholecystectomy. Although the latter were at higher risk (higher APACHE II scores), their outcome except for longer intensive care unit stays, was not different from laparoscopically treated patients. Lack of superiority of laparoscopic over open cholecystectomy in the present study seemed due to clinical characteristics of AC in geriatric patients which may lead to late diagnosis and treatment. Preoperative ERCP by further delaying surgery may contribute to loose any potential benefit of an early laparoscopic procedure. The place of preoperative ERCP and the timing of LC in geriatric patients with AC therefore may need to be redefined. PMID- 11868507 TI - Intra-abdominal metastases from primary carcinoma of the lung. AB - The case of a 56-year old male with an intra-abdominal metastasis from a primary lung cancer is presented. He was admitted for abdominal obstruction, toxic syndrome and paraumbilical pain. He had a previous history of squamous cell carcinoma of the right lung, for which he had undergone a right upper lobectomy in 1995, four years prior to the development of the abdominal obstruction. A debulking operation and bowel resection for the intestinal metastasis was performed. Eleven months after this operation the patient developed a recurrence: he underwent another debulking operation with resection of the sigmoid colon, jejunal segment and a small part of the bladder. The patient is alive and well 13 months after the initial operation. Intra-abdominal metastases of bronchial carcinoma may be observed with greater frequency, because of the improved survival of the patients with lung cancer. Metastatic small bowel carcinomas are rare and should be considered in the differential diagnosis of acute abdominal syndromes of patients with known history of the lung cancer. Bowel resection and debulking of the metastatic tumour mass give the best palliation and improve short-term survival. PMID- 11868508 TI - Chronic osteomyelitis after sternotomy. AB - Two patients with chronic sternal osteomyelitis after an initially uncomplicated coronary artery bypass grafting (CABG) operation are described. Chronic osteomyelitis, caused in both cases by Pseudomonas aeruginosa, occurred six and four months after CABG respectively. Because chronic infection failed to respond to local wound care and medical therapy, more radical treatment was needed. Steel wires were removed and surgical debridement was performed. In one patient, an additional omental transposition was performed. In both cases radical debridement in combination with antibiotics successfully eradicated the infection. PMID- 11868509 TI - Thrombosis of an aneurysm of the basilic vein upper extremity venous aneurysm. AB - We present the case of a large venous aneurysm in the upper extremity of a 71 year-old male. The patient presented with acute thrombosis of the aneurysm, which was only diagnosed at operation. Aetiology, diagnosis and treatment of this uncommon lesion are discussed. PMID- 11868510 TI - Round ligament varicosities mimicking inguinal hernia: a diagnostic challenge during pregnancy. AB - Groin swelling first evokes inguinal or femoral hernia but many other conditions may account for it. We describe varicosities of round ligament in a 27-year-old pregnant woman. She presented with a groin mass mimicking an inguinal hernia. Diagnosis was made during surgical exploration. This case report strengthens the fact that varicosities of the round ligament, favoured by hormonal and mechanical factors, should be evoked in a pregnant woman complaining of a groin mass. Ultrasonographic examination of the groin should be performed in such cases to avoid unnecessary surgery. PMID- 11868512 TI - A history of the Wisconsin Surgical Society. PMID- 11868511 TI - Primary torsion of the greater omentum. AB - Primary or idiopathic greater omental torsion remains a rare cause of acute surgical abdomen in adults and children. The aetiology is as yet unknown and the treatment of choice, once diagnosis is established, is resection of the torted omentum. We report our experience with three such cases encountered over the last five years, two of which were diagnosed and subsequently managed laparoscopically. The performance of diagnostic laparoscopy for acute abdominal pain of an undetermined origin may lead to an increased detection of this condition and subsequent therapeutic intervention. PMID- 11868513 TI - Breaking bad news and obtaining do not resuscitate orders. PMID- 11868514 TI - End-of-life care: learnings over a lifetime. PMID- 11868515 TI - Making end-of-life care easier for physicians. PMID- 11868516 TI - Accepting death. PMID- 11868517 TI - Factors influencing rural Wisconsin elders in completing advance directives. AB - PURPOSE: Advance directives can assist in end-of-life decision-making. This study explores the factors that influence completion of advance directives. METHODS: Qualitative study using in-depth interviews of 10 patients over age 60 in a rural office practice. RESULTS: Major themes: (1) patient autonomy: the patient and family role as decision maker, the physician as educator; (2) barriers that deter patients from completing advance directives include fear and confusion about these documents; and (3) quality of life is an important value in end-of-life decision-making. CONCLUSIONS: Decision making about advance directives is a personal and family issue, but physician efforts to initiate the process and educate patients are important, especially if done routinely in the office or preferably in community settings. The main difference between those who do and don't complete advance directives is an understanding of the documents and their purpose. PMID- 11868519 TI - Physicians and the pharmaceutical industry: a growing embarrassment and liability. AB - The pharmaceutical industry, comprising corporate-owned, for-profit enterprises, employs nearly one sales representative with a budget of about $100,000 for each 11 practicing physicians. Physician contact with these individuals, epitomized by the drug company "free lunch," results in non-rational prescribing and increases the overall cost of prescription medication to patients. Other influential methods employed by the industry include sponsorship of tainted journal articles, restrictions on research, and biased continuing medical education. Physicians are often unaware of their vulnerability to these tactics. Remedies are suggested. PMID- 11868518 TI - Nutritional assessment of a predominately African-American inner-city clinic population. AB - BACKGROUND: Dietary changes are recommended as first-line treatment and prevention strategies of chronic diseases. The African-American diet in particular has been characterized as high in sodium and fat while lacking in fiber. METHODS: Using a 24-hour diet recall with a modified food frequency questionnaire (FFQ), we assessed the diet/nutritional status of patients attending 2 inner-city health centers in Milwaukee. Point-of-purchase data on food sales was collected for comparison. RESULTS: Diets were high in fat (42.7% of total calories), saturated fat (14.6%), and sodium (nearly 150% of the daily recommended value), while lacking in dietary fiber (30% of the daily recommended value). Point-of-purchase data correlated with dietary intake. CONCLUSIONS: The diets of the population surrounding these inner-city health centers were high in fat and sodium, but low in fiber. Programs are needed to address the unhealthy dietary practices within this population. PMID- 11868520 TI - Firearms injury center's objectivity questioned. PMID- 11868521 TI - A promise for palliative care at the end-of-life. PMID- 11868522 TI - Make pneumococcal immunization a priority. PMID- 11868523 TI - Dilemmas in geriatrics: scenario 1 results. PMID- 11868524 TI - [Laser irradiation in the treatment of ischemic heart disease]. AB - Cardiodynamic changes due to beta-blocker carvedilol and low-intensity infrared laser radiation were compared in 115 patients with ischemic heart disease (IHD). The comparison has shown a similar positive effect on heart contractility and diastolic function. This gave arguments for feasibility of laser beam usage as a neurohormonal modulator in IHD patients to reduce cardiac remodulation and prevent cardiac failure. PMID- 11868525 TI - [Spectral analysis of the variability of heart rhythm in the analysis of changes in the autonomic regulation during treatment of hypertension with sodium chloride baths]. AB - Changes in spectral parameters of heart rate variability were assessed in patients with mild and moderate hypertension (n = 48) and healthy subjects (n = 34) taking sodium chloride bath and in active orthostatic test before and after balneotherapy. Significant changes were not registered. The course of balneotherapy stimulated sympathetic or parasympathetic activity (relative enhancement occurred in 67, absolute in 15% versus 8 and 10% patients, respectively). PMID- 11868526 TI - [Chest cryomassage in the treatment of patients with chronic obstructive bronchitis]. PMID- 11868527 TI - [Changes in the autonomic status in vegetovascular dystonia syndrome during exposure to permanent magnetic field ]. AB - Permanent magnetic field was applied by placing two magnetic discs (induction 60 mT) in the projection of great arteries on both sides symmetrically. The treatment was given to hypertensive patients with vegetovascular asthenia. A course of such magnetic treatment affects pathogenetic mechanisms of vegetative disorders due to the ability of permanent magnetic field to restore vegetative homeostasis, normalize sympathicotony and vegetative regulation. PMID- 11868529 TI - [Changes in the functional state of the neuro-muscular apparatus in patients with distal diabetic polyneuropathies after treatment with interference currents and "yellow" turpentine baths]. PMID- 11868528 TI - [The use of extremely high frequency electromagnetic fields during acute period of ischemic stroke]. AB - The rheological, coagulatory and clinical examination of 70 patients with acute ischemic stroke has demonstrated that EHF therapy (53.53 GHz) improves rheological and coagulatory parameters in such patients, has a positive effect on hemostasis and clinical course of acute ischemic stroke. The technique and doses are presented. PMID- 11868530 TI - [Physical therapy in the treatment of myofascial pain syndrome of the chest]. AB - Differentiated combined treatment of myofascial pain syndrome of the chest has been performed in 126 patients. The treatment included soft tissue manual therapy, therapeutic exercise and physiotherapy. Physiotherapeutic techniques varied with the stage of the exacerbation. As shown by neuroorthopedication, the treatment resulted in significant inhibition of the activity of the trigger points. The remission lasted 384.2 +/- 41.3 days. PMID- 11868531 TI - [Balneotherapy of children with complications of craniocerebral injuries]. AB - Children with aftereffects of craniocerebral trauma have received sanatorium treatment including exercises, massage, mineral baths of different chemical composition. Adequate therapeutic measures taken in due time diminish the number of the aftereffects and their severity. PMID- 11868532 TI - [Possible use of balneotherapy for atherosclerosis prevention]. AB - Balneotherapy was assessed as a method of primary prevention of atherosclerosis. The effectiveness of effervescent hydrocarbonate magnesium-calcium water was evaluated by modification of structural-functional state of red cell membrane in examinees with risk factors. It is shown that combined balneotherapy remodels abnormal lipid component of cellular membranes. This allows using this method in primary prevention of atherosclerosis in persons with risk factors in the absence of changes in serum lipid spectrum. PMID- 11868533 TI - [Keratinocyte immunophenotypes in patients with psoriatic arthritis and their changes after treatment with deresinated naphthalan]. AB - The study of phenotyping features, membrane markers of epidermic keratinocytes, their production of adhesive molecules has established structural basis of favourable effect of deresinated naphthalan on the skin and articular syndromes in patients with psoriatic adtheritis. The immunohistochemical examination of the skin biopsies using monoclonal antibodies in the test of indirect immunofluorescence has found that deresinated naphthalan in patients suppresses immunoinflammatory reactions in the skin. Monitoring of expression of molecules HLA--on keratinocytes may be helpful for prediction of the skin and joint manifestation. PMID- 11868535 TI - [Novel approaches to dissemination of innovations in the field of rehabilitation and health resort medicine]. PMID- 11868534 TI - [Balneological use of bath concentrate containing xidifon and essential oils from ginger, nutmeg, and sandal]. PMID- 11868537 TI - [Improvement of dissemination of novel medical technologies into health resort practice]. PMID- 11868536 TI - [Certification of physiotherapists]. PMID- 11868538 TI - [Various indices of biological activity of the peat mud from the "Varzi-Iatchi" health resort]. PMID- 11868539 TI - [Cervical myelopathy and its treatment with physical factors]. PMID- 11868541 TI - [Rehabilitation of patients with purulent bronchopulmonary complications]. PMID- 11868540 TI - [Lipid peroxidation and antioxidant protection during hydrogen sulfide balneotherapy in children with primary arterial hypertension]. PMID- 11868542 TI - [Cosmetic procedures in health resort treatment]. PMID- 11868543 TI - [Early specialization]. PMID- 11868544 TI - [The effect of personality traits on rehabilitation of patients with ischemic heart disease after aortocoronary bypass]. AB - The course of ischemic heart disease (IHD) was studied in patients who had undergone coronary artery bypass operation with consideration of types of emotional and behavioral activity. A total of 83 IHD patients were examined using Jenkin's questionnaire. Clinical and psychophysiological characteristics of patients with different behavioral types are described. Significant differences in reactivity of cardiovascular system were found. Behavioral type should be taken into consideration in planning differentiated care for such patients. PMID- 11868545 TI - HIV testing & treatment of children. AB - In the last year, courts in Canada, the US, and the UK have decided cases in which child welfare authorities have disputed parents' decisions to refuse HIV testing or treatment for their children. This article reviews recent US developments regarding refusal of treatment, and reports the outcome of a recent UK decision compelling the HIV testing of a minor against her parents' wishes. It then provides an update on two recent Canadian cases concerning a parent's refusal to treat an HIV-positive child with antiretroviral therapy, and a brief overview of the current state of Canadian law on the issue of minors and medical treatment. PMID- 11868546 TI - HIV/AIDS and children's rights. AB - Today, the majority of all new HIV infections occur among children and young people under 25 years of age, the people who were born and who have grown up during the AIDS epidemic. The epidemic is straining resources in already impoverished communities and creating new obstacles to the realization of children's rights to survival, development, and protection. The failure to ensure children's rights creates opportunities for HIV infection; at the same time, HIV/AIDS creates opportunities for the violation of children's rights. Advances in the realization of children's rights, including the implementation of the United Nations Convention on the Rights of the Child (UNCRC), are necessary to stem the growth of the AIDS epidemic. We reproduce a fact sheet produced by the Interagency Coalition on AIDS and Development (ICAD) that summarizes available information on HIV/AIDS and children, and discusses the effect of HIV/AIDS on children's rights. The fact sheet also provides information about other, essential resources on HIV/AIDS and children. PMID- 11868547 TI - AIDS organizations denied leave to intervene in insurance case. AB - In October 1999, the BC Court of Appeal denied the request of the Canadian AIDS Society (CAS) and the BC Persons with AIDS Society (BCPWA) to intervene in the appellate hearing of a case raising questions about the responsibilities of employers and employees to preserve entitlement to life insurance coverage. In this case, the issue of the mental capacity of a man with AIDS-related dementia was a key issue. PMID- 11868548 TI - No removal of HIV-positive man for marijuana conviction. AB - In June 1999, the Immigration Appeal Division of the Immigration and Refugee Board dismissed a motion by the federal government to order the removal from Canada of an HIV-positive (and hepatitis C-positive) man for a drug offence. PMID- 11868549 TI - Criminalization of assisted suicide challenged. AB - In September 1999, Jim Wakeford, a Toronto man with HIV/AIDS, launched a civil action against the Attorney General of Canada, challenging the constitutionality of sections of the Criminal Code that criminalize assisted suicide. PMID- 11868550 TI - No duty to preserve benefits for employee with dementia. AB - In August 1999, an Ontario trial court granted an employer's motion for a summary judgment dismissing a claim against it by the estate of a former employee who died of AIDS-related illness. PMID- 11868551 TI - Appeal board denies public health insurance coverage to immigrants on Minister's Permits. AB - In a decision released in August 1999, the Health Services Appeal Board in Ontario upheld the decision to deny provincial health coverage to an HIV-positive woman who had been denied permanent resident status on the basis of "medical inadmissibility" but was living in Canada on an Immigration Minister's Permit issued on "humanitarian and compassionate grounds." PMID- 11868552 TI - Newfoundland man pleads guilty to criminal harassment. AB - In May 1999, a Newfoundland trial court sentenced a 30-year-old man after he pleaded guilty to one count of criminal harassment for posting public notices that his ex-girlfriend was HIV-positive and had unprotected sex without disclosing this fact. PMID- 11868553 TI - Allegations of HIV infection raised in criminal trial of escort agency owner. AB - At the beginning of October 1999, a Toronto man who owned an escort agency went to trial in the Ontario Superior Court of Justice on numerous charges of living off the avails of prostitution and procuring for the purpose of prostitution. On the first day of trial, the prosecutor indicated he would be introducing evidence suggesting that three of the women who worked as escorts for Mark Lucacko's agency may be HIV-positive. PMID- 11868554 TI - A different kind of risk?--pregnant women's experience of HIV testing in pregnancy. AB - This paper describes some emerging themes developed from conversations with pregnant women participating in a pilot study in Ottawa and Montreal. These initial findings will inform a national study that will address the issue of HIV screening in pregnancy from the perspectives of the women themselves. Through an iterative series of conversations with each woman, the women in this pilot shared their experiences of being offered and of completing HIV testing during their pregnancy. The paper addresses the specific question of whether, in the context of promising new interventions to reduce perinatal transmission and rising levels of HIV infection among Canadian women of child-bearing age, the drive to screen as many pregnant women as possible has resulted in an abrogation of a woman's right to the established principles of HIV testing. Specifically, what is the evidence from the women in this pilot of adherence to the principles that testing for HIV should always be voluntary and should always be carried out only after the person has given their informed specific consent? PMID- 11868556 TI - Legal, ethical, and human rights issues facing East and Southeast Asian Canadians. AB - In the last issue of the Newsletter, we reported about a project on HIV/AIDS and discrimination in South Asian communities in Canada, undertaken by the Toronto based Alliance for South Asian AIDS Prevention (ASAP). In this issue, we report about another project focusing on issues of discrimination in ethnocultural communities in Canada: a project on legal, ethical, and human rights issues facing East and Southeast Asian-Canadians, undertaken by a Toronto-based AIDS service organization, the Asian Community AIDS Services (ACAS). The project concluded that "societal discrimination, systemic service deficiency and resource allocation barriers contribute to a hostile environment that challenges the self esteem, sense of efficacy, health and well being of Asians infected and affected by HIV/AIDS." PMID- 11868555 TI - Discrimination and human rights abuse in Russia. AB - AIDS-related discrimination and human rights violations in Russia are widespread. The problems have been documented in a report prepared by the Russian NAMES Foundation, a national non-profit organization that provides services in HIV/AIDS awareness, offers social and psychological support to people affected by the AIDS epidemic, and advocates effective and non-discriminatory AIDS policies. The report is entitled AIDS-Related Violations of Human Rights and the Russian Legislation. The information presented in the report is based on testimonies by people with HIV/AIDS, their friends and families, medical professionals, state officials, workers at non-governmental organizations, and other citizens. The following is an edited version of the report. PMID- 11868557 TI - The International Guidelines on HIV/AIDS and Human Rights--three years on. AB - 1999 was the third anniversary of the drafting of the International Guidelines on HIV/AIDS and Human Rights. This article reviews the history of the Guidelines, notes some obstacles to their implementation, and concludes with suggestions for governments and development agencies. PMID- 11868558 TI - Developing effective HIV prevention programs for inmates: results from an Ontario wide survey. AB - Many studies of the characteristics, behaviours, and prevalence of HIV among those admitted to Canadian correctional facilities indicate that inmates are at increased risk for HIV infection and that they be targeted for HIV interventions. Yet the development and implementation of these programs has been much too slow and, at times, inappropriate to the prison environment. Further, the effectiveness of current and proposed policies and programs must be evaluated to ensure that the goal of reducing HIV transmission is achieved. To address these issues, a team of University of Toronto researchers undertook a study entitled "The Social and Structural Determinants of HIV-Related Risk Behaviours among Prisoners: Implications for Prevention." This study surveyed adult males and females incarcerated in six provincial correctional centres in Ontario. The following provides a brief overview of the study and its results. PMID- 11868559 TI - Prisoner settles case for right to start methadone in prison. AB - In July 1999, Dwight Lowe, an inmate at Kent Institution previously using heroin, settled his case against Correctional Service Canada (CSC) in which he challenged as unconstitutional CSC's refusal to permit him to initiate methadone maintenance treatment while in prison. PMID- 11868560 TI - Ethical review in community-based HIV/AIDS research. AB - AIDS Vancouver recently released a report on ethical review in community-based research. The report identifies some barriers facing community-based researchers and makes recommendations for facilitating this area of HIV/AIDS research in Canada. The following summary of the report was provided by its author. PMID- 11868561 TI - The University of Ottawa Community Legal Clinic. AB - This is the third in a series of articles about legal clinics that provide specialized services to people with HIV/AIDS. In previous issues of the Newsletter, Ruth Carey provided an overview of the work of the HIV & AIDS Legal Clinic (Ontario) (HALCO), and Johanne Leroux described the work of the legal clinic of the Quebec Committee for people with HIV in Montreal. In this article, Jennifer Duff, a low student and caseworker at the University of Ottawa Community Legal Clinic, describes the work of the HIV Legal Services division of the Ottawa Clinic. PMID- 11868562 TI - Criminal law and HIV/AIDS: update II. AB - This article reviews new developments in the area of criminal prosecutions for HIV transmission or exposure, or developments that have come to our attention since the last issue of the Newsletter. This column will continue to be a regular feature. However, we will not continue to report cases or legislation from jurisdictions other than Canada unless they represent a significant development, either in this area of the law or for the jurisdiction in question. PMID- 11868563 TI - BC human rights tribunal rules on discrimination by London life. AB - In June 1999, a British Columbia human rights tribunal ruled that London Life Insurance Company had violated the province's Human Rights Act in 1994 by refusing to sell life insurance (on his own life) to an HIV-negative man on the basis that his wife is HIV-positive. PMID- 11868564 TI - Critique of Podiatric biomechanics by William Eric Lee, DPM. PMID- 11868565 TI - Vital pulp therapy--a review. PMID- 11868566 TI - The ticket to work. PMID- 11868567 TI - Clinical quiz. Chronic granulomatous disease. PMID- 11868568 TI - Ferguson v. City of Charleston, S.C. PMID- 11868569 TI - Bone marrow transplantation. PMID- 11868570 TI - Trauma surgery. PMID- 11868571 TI - Pemberton v. Tallahassee Memorial Regional Center. PMID- 11868572 TI - [Further investigation on Second Volume of Essay on Simple Recipe and Pulsology (SESR) and Wang's Simple Recipes (WSR)]. AB - There are 2 extant editions of SESR, written in Chunyou Jiachen (1244) reign. In the facsimile copy of Song manuscript recorded in Kojima's Records of Investigating Ancient Classics which was brought by Yang Shoujing and is now collected in the Library of Palace Museum in Taipei. There is also a facsimile copy of Song dynasty postscribed by Taki Genkan now collected in Library of Relics and Naturalist Academy of National Imperial Secretariat in Japan. The book is written in one volume, dealing first with four diagnostic methods, followed by drug and therapy and compounding of recipes and "processing" of drugs, thus forming a complete system. The WSR included in Classified Collection of Recipes is not the Essay on Pulsology, neither WANG Shou's Simple Recipes, but Yi jian gui yi (Unity of Simplicity and Easiness) by XU Ruoxu of Yuzhang, Jiangxi of Yuan dynasty, being an epitomized work of all physicians of Yongjia medical schools with only 3 pieces of incomplete papers remained. PMID- 11868574 TI - [Effects of 6-degree head-down bed rest on physical fitness]. AB - We investigated changes in enduring physical fitness during a ground-based weightlessness simulation of 6-degree head-down bed rest (HDBR). Ten healthy volunteers participated in the 6 days of HDBR. In each, their physique, muscular strength, physical fitness, cardiovascular function, and hematocrit were measured before and after HDBR. The results are as follows: (1) Thigh extensor power: the cross-sectional areas of the thigh and leg muscles were significantly decreased after HDBR, but recovered immediately afterwards. (2) Hand grip power and instantaneous power: the body response time showed no significant change before and after HDBR. (3) High physical capacity was correlated with a greater decrease in physical capacity after HDBR. (4) Remarkable functional depression appeared in the cardiovascular system compared with the gas exchange system as measured by the enduring physical fitness test. (5) The functional depression in the cardiovascular system lasted for more than 3 days after HDBR. (6) Six days of HDBR had little influence on physical fitness attributable to the inactivity and seemed to effectively simulate weightlessness for the cardiovascular system. PMID- 11868575 TI - Gentamicin: effect on E. coli in space. AB - Previous investigations have shown that liquid bacterial cultures grown in space flight were not killed as effectively by antibiotic treatments as were cultures grown on Earth. However, the cause for the decreased antibiotic effectiveness remains unknown. Possible explanations include modified cell proliferation and modified antibiotic transport in the culture medium. Escherichia coli cultures were grown in space flight (STS-69 and STS-73), with and without gentamicin, on a solid agar substrate thus eliminating fluid effects and reducing the unknowns associated with space-flight bacterial cultures in suspension. This research showed that E. coli cultures grown in flight on agar for 24 to 27 hours experienced a heightened growth compared to simultaneous controls. However, addition of gentamicin to the agar killed the bacteria such that both flight and ground control E. coli samples had similar final cell concentrations. Therefore, while the reported existence of a decrease in antibiotic effectiveness in liquid cultures remains unexplained, these data suggest that gentamicin in space flight was at least as effective as, if not more effective than, on Earth, when E. coli cells were grown on agar. PMID- 11868576 TI - Unsupervised classification of Space Acceleration Measurement System (SAMS) data using ART2-A. AB - The Space Acceleration Measurement System (SAMS) has been developed by NASA to monitor the microgravity acceleration environment aboard the space shuttle. The amount of data collected by a SAMS unit during a shuttle mission is in the several gigabytes range. Adaptive Resonance Theory 2-A (ART2-A), an unsupervised neural network, has been used to cluster these data and to develop cause and effect relationships among disturbances and the acceleration environment. Using input patterns formed on the basis of power spectral densities (psd), data collected from two missions, STS-050 and STS-057, have been clustered. PMID- 11868577 TI - Medicare program; fee schedule for payment of ambulance services and revisions to the physician certification requirements for coverage of nonemergency ambulance services. Final rule with comment period. AB - This final rule establishes a fee schedule for the payment of ambulance services under the Medicare program, implementing section 1834(l) of the Social Security Act. As required by that section, the proposed rule on which this final fee schedule for ambulance services is based was the product of a negotiated rulemaking process that was carried out consistent with the Federal Advisory Committee Act and the Negotiated Rulemaking Act of 1990. The fee schedule described in this final rule will replace the current retrospective reasonable cost payment system for providers and the reasonable charge system for suppliers of ambulance services. In addition, this final rule requires that ambulance suppliers accept Medicare assignment; codifies the establishment of new Health Care Common Procedure Coding System (HCPCS) codes to be reported on claims for ambulance services; establishes increased payment under the fee schedule for ambulance services furnished in rural areas based on the location of the beneficiary at the time the beneficiary is placed on board the ambulance; and revises the certification requirements for coverage of nonemergency ambulance services. PMID- 11868578 TI - Crossing the boundary to the third millennium AD. PMID- 11868579 TI - Role expansion in intensive care: survey of nurses' views. AB - The study described here is an investigation of intensive care unit (ICU) nurses' perceptions of how their role is expanding. Six specific skills were selected for consideration. A questionnaire was distributed to nurses working in ICUs in three different hospitals. Questions were based around issues such as education and assessment policies, the transfer of work between nurses and other groups of workers and the effects of ICU nurses accumulating additional skills. Completed questionnaires were received from 33 participants (68.75% response rate). The results showed that most of the nurses were carrying out the skills suggested. The training provided to underpin skill development was a balance of teaching and supervision. However, assessment practices varied. The majority of nurses felt that the existing training programmes were adequate. Data revealed that a range of duties could be released from their role and taken over by other workers, such as technicians and health care assistants. Participants suggested that the crucial element of the nursing role was the incorporation of many activities into the provision of continuous one-to-one total patient care. This was the aspect of work that was reported as being the most satisfying. It was perceived that patients benefited from nurses extending their role in the areas discussed. PMID- 11868580 TI - Patients' perspective of quality of care in a high-dependency unit. AB - This study was designed to identify and explore the patients' perspective of the concept quality of care within the context of a high-dependency unit (HDU). Data were collected in two phases. In phase one, 55 patients were interviewed to clarify the concept quality of care. In phase two, 77 patients were interviewed to explore this concept in further detail. This study has clarified the patients' concept of quality of care into the principal constructs of the environment, meeting individual needs, staff attitudes and manners, organization of care, communication, staff skills and family and friends. It has been successful in providing patients with a voice to comment upon an aspect of service provision. A well-known concept has been clarified and applied to a high-dependency environment. This has identified the need for future evaluation of HDUs to go beyond traditional physiological measures, to incorporate the service experience itself and the impact it has upon the patients. PMID- 11868581 TI - Intravenous therapy in critically ill adults: developing a clinically and cost effective approach. AB - Intravenous therapies are the most common intervention for critically ill adults. Using a systematic approach as described by Droogan and Song (1996), a review of the literature was undertaken to determine whether the frequency of changing intravenous administration sets in critically ill adults with central venous catheters (CVCs) affected the incidence of CVC-related sepsis/systemic inflammatory response syndrome (SIRS)/bacteraemia. Two major randomized controlled trials were included in the review (Maki et al. 1987; Snydman et al. 1987), which conclude that increasing the change frequency of administration sets from 24 to 72 hours does not significantly increase the incidence of sepsis. This can therefore lead to considerable cost savings as well as ensuring clinically effective care. The review criteria excluded a plethora of related studies. However, these studies do corroborate the findings of Maki et al. and Snydman et al. They are summarized in the tables and are taken into account when making recommendations for clinical practice and future research. Clinical practice guidelines which are being implemented and evaluated locally are offered for the reader's consideration. PMID- 11868582 TI - Exploratory study of nursing in an operating department: preliminary findings on the role of the nurse. AB - This study was exploratory and describes how nursing was viewed and practised by nurses who worked in an operating department. It also highlighted factors that might influence the role performance of operating department nurses. The research involved interviews with a sample of 6 nurses working in an operating department, observation of 32 hours of nursing work over 6 operating sessions, in addition to the analysis of various documents, including the nursing care plans of 22 patients. Data were triangulated and analysed by constant comparison. Findings indicated that nurses had difficulty in articulating exactly what it was that operating department nursing entailed, but rather viewed their role in terms of the functions they performed. Observations indicated that the nursing role was primarily orientated toward the physical rather than the psychological aspects of care-giving. Furthermore, it appeared that the medical profession, nursing philosophy/leadership and the characteristics of patients all influenced the manner in which nurses enacted their role. These findings suggest that further research into the role of the nurse within the operating department environment is warranted. Key factors from this study were developed into a framework suitable for guiding future study of the nursing role in this environment. PMID- 11868583 TI - Utilization of the Iowa Model in establishing evidence-based nursing practice. AB - Clinical practice based on tradition or established rituals appears to be widespread amongst a variety of nurses and practice settings. However, tradition based practice may not necessarily be based on sound scientific evidence and could potentially be harmful to patients or result in inappropriate utilization of resources. Conversely, evidence-based practice is the utilization of the best available empirical evidence in the practice setting, to facilitate sound clinical decision-making. Suctioning ventilated patients is a necessary and important aspect of patient care. However, normal saline instillation prior to suctioning, in order to loosen secretions, remains a common nursing procedure despite research suggesting that there is no clear benefit and in some instances may be harmful. Several models have been developed over the past few years to facilitate nursing practice that is based on research or the best available evidence. The Iowa Model, developed at the University of Iowa Hospitals and Clinics, serves as a framework to improve patient outcomes, enhance nursing practice and monitor health care costs. Moreover, it facilitates the application of empirical evidence to clinical practice. This paper will discuss the utilization of the Iowa Model to promote evidence-based nursing practice, with regard to normal saline instillation prior to suctioning, in the critical care unit of a 100-bed hospital in Hong Kong. Patient, staff and fiscal outcomes will also be reported. PMID- 11868584 TI - Zanamivir (Relenza )--a new treatment for influenza. AB - Deaths from influenza are recorded every year in the UK and flu epidemics raise concerns for the safety of the young, the elderly and other vulnerable groups in the community. Not surprisingly, the launch of zanamavir (Relenza) by Glaxo Welcome (Middlesex, UK) has received much attention in the medical media and lay press. Just who should receive this treatment and under what conditions it should be given remains to be confirmed in practice. PMID- 11868585 TI - Optimizing oxygen delivery: haemodynamic workshop. Part 1. PMID- 11868586 TI - Professional journalling over time: position of the inside nurse-researcher in intensive care. AB - The purpose of this paper is to examine the use of professional journalling in an intensive care unit. In particular, this paper explores the power and contextual relations that shaped nurse-nurse and nurse-doctor decision-making, from the author's perspective as the inside nurse-researcher. A variety of research methods were used for this study, including professional journalling, participant observation, and individual and focus group interviews. The author also worked as a clinical nurse specialist, as the inside nurse-researcher, in the unit under investigation. The results indicated that the power and contextual relations constructing nurse-nurse and nurse-doctor decision-making focused on two major issues. These concerned: drawing on particular forms of knowledge and acquiring differential visibility by the nurse in decision-making. The study demonstrated that professional journalling is an effective research method for helping nurses to question decision-making critically, not for examining the position of an inside nurse-researcher, and for supporting data obtained from other research methods. PMID- 11868588 TI - Nurse consultant--a role whose time has come. PMID- 11868587 TI - Palivizumab (Synagis) for prevention of respiratory illness in children. AB - Respiratory syncytial virus commonly causes infections that are of little consequence to most of us. However, for the newborn and at-risk older children, infection can mean serious, even life-threatening respiratory illness affecting pulmonary development and resulting in chronic chest disease. Palivizumab is a new medicine of the high-tech era designed specifically to prevent such problems. Its managed entry into clinical practice sets a challenge for health authorities across the country in ensuring most cost-effective delivery of health care. PMID- 11868589 TI - The Audit Commission Report (1)--Creating a 'data collecting' culture in critical care. PMID- 11868590 TI - Carers' opinions and emotional responses following cardiac surgery: cardiac rehabilitation implications for critical care nurses. AB - The recent joint position statement made by the RCN Critical Care and Rehabilitation Nursing Forums highlights the need for rehabilitation to commence early. This paper reports the findings of a descriptive survey of 59 carers of cardiac surgery patients and presents implications for cardiac rehabilitation. Data were obtained by postal questionnaire during early recovery (one week following discharge) and six weeks later. The questionnaire explored carers' perceptions about the timing of discharge from hospital; opinions of the information provided by hospital staff; and anxiety and depression measured on the Hospital Anxiety and Depression Scale. The results indicated that carers assumed a heavy burden once the patient had left the specialist cardiac centre. Carers responding at one week were less satisfied with the timing of discharge than those questioned at six weeks. Information provided by nurses was rated more highly than that provided by doctors or physiotherapists. However, there was scope for increasing input. The findings suggest that cardiac rehabilitation needs to be aimed at carers as well as patients. Investment in targeted carer support could facilitate patient recovery and rehabilitation. Strategies aimed at the carer need to begin early and commence during the acute stage of the patients' recovery. PMID- 11868591 TI - Intensive care nurses' experiences of caring. Part 1: Consideration of the concept of caring. AB - Curiosity as to what other intensive care nurses experienced as caring practice in a high-tech environment such as intensive care was prompted by a dilemma that arose in the author's own clinical practice. One consequence was this study which took place in a 12-bedded intensive and coronary care unit (ICU). Although there is a vast body of literature discussing caring in nursing, little is related to the intensive care environment. The first part of this paper contains discussion of the concept of caring related to this aspect of nursing, thus addressing the initial stages of the research process. This was guided by the research question 'What is caring?' Part two of this paper will present the phenomenological research study designed to answer this question. PMID- 11868592 TI - Optimizing oxygen delivery: haemodynamic workshop. Part 3. PMID- 11868593 TI - Health-related quality of life of ICU survivors: review of the literature. AB - The importance of health-related quality of life (HRQL) as a relevant outcome measure for patients requiring intensive care unit (ICU) management has only recently been recognized. A better understanding of how this expensive service affects the health and well-being of its survivors will allow nurses and other health care professionals to plan for and provide appropriate follow-up care. This paper contains a review of the theoretical basis for quality-of-life measures, discussion of some of the methodological issues, and examination of the findings from recent studies of the quality of life of ICU patients. Although not conclusive, the review identified that ICU survivors generally have poorer HRQL scores after a 6-12 month period of recovery than during their pre-admission period. This cohort also have lower HRQL than the age-adjusted general population. Despite this apparent poorer state of health, participants often claimed to be satisfied with their HRQL. Methodological weaknesses were evident in the studies reviewed, Challenges in terms of recruitment and retention of subjects, instrumentation and data collection became evident from this review. Continued research in this area is recommended and should address the weaknesses identified. PMID- 11868594 TI - Are manual gestures, verbal descriptors and pain radiation as reported by patients reliable indicators of myocardial infarction? Preliminary findings and implications. AB - Patients experiencing an episode of acute chest pain need to be assessed promptly and effectively to ensure optimal management. The aim of this study was to investigate whether there were specific aspects of patients' symptom reports which could be viewed as additional indicators of myocardial infarction (MI) and contribute to the assessment process. The sample consisted of 267 patients who presented with an episode of acute chest pain. Methods of data collection were based on, or modified from, previous studies which had investigated the use of manual gestures, choice of verbal descriptors and extent of pain radiation in patients with and without MI. To determine whether these variables were of significance in the diagnosis, the responses of those with (n = 118) and without MI (n = 149) were compared. The results suggest that it is currently impossible to draw any conclusions as to whether the variables studied can be judged as reliable indicators of MI. The findings indicate that there are some differences between the groups particularly in the language used. Moreover, women with MI characterized their symptoms through stronger emotive words such as 'worrying' (P = 0.014) 'frightening' and 'intolerable' and also differed from their male counterparts in their reports regarding pain radiation. Implications for practice and for research are discussed. PMID- 11868595 TI - Frequency and pathogenesis of silent subcortical brain infarction in acute first ever ischemic stroke. AB - OBJECTIVES: We have often observed silent subcortical brain lesions on CT or MRI in first-ever ischemic stroke, but there is little published information on the relationship of these lesions to stroke subtypes. Here, we describe the incidence of MRI-detected silent subcortical brain lesions, including infarctions and white matter lesions, in a series of patients with first-ever ischemic stroke classified according to stroke subtypes. We also discuss the pathogenesis of these silent subcortical lesions. PATIENTS: We evaluated 171 patients with acute first-ever ischemic stroke. METHODS: The subjects were divided into three groups: lacunar, atherothrombotic and cardioembolic infarction groups. We evaluated silent subcortical brain infarction (SSBI), enlargement of perivascular space (EPS), and other white-matter lesions using MRI. RESULTS: Hypertension was observed in 67.6% of lacunar infarction, 57.1% of atherosclerotic infarction, and 54.1% of cardioembolic infarction. SSBI was more frequently observed in lacunar infarction than the others (lacunar vs. atherothrombotic vs. cardiogenic infarction, 81.5% vs. 44.4% vs. 42.1%, p=0.006). High-grade EPS (grade 2 or higher) was also observed more frequently in lacunar infarction than in the others (lacunar vs. atherothrombotic vs. cardiogenic infarction, 63.3% vs. 24.2% vs. 0%, p<0.001). Scheltens' score of silent subcortical lesions was significantly higher in lacunar infarction than in the others. CONCLUSIONS: The frequency of silent subcortical ischemic brain lesions was significantly higher in lacunar infarction than in atherosclerotic or cardioembolic infarction. We suggest that the pathogenesis of silent subcortical ischemic brain lesions is common to that of lacunar infarction, that is, small-vessel vasculopathy. PMID- 11868596 TI - Severe pulmonary hypertension in a patient with systemic lupus erythematosus and minimal lupus activity. AB - Pulmonary hypertension (PH) sometimes occurs in patients with systemic lupus erythematosus (SLE). We report a case of 51-year-old-woman with PH associated with SLE. She had been diagnosed as SLE on the basis of pericardial effusion, hematological disorder, positive antinuclear antibody, and hypocomplementemia. Despite minimal lupus activity, she had marked elevation of pulmonary arterial pressure (101/53 mmHg) and decreased cardiac index (1.5 l/min/m2). Symptoms related to PH were progressive under treatment with oral corticosteroids, oxygen, calcium antagonists, and warfarin. After 17 months of epoprostenol treatment, she died of pulmonary infarction. SLE-associated PH is often severe and progressive even in association with minimal activity. PMID- 11868597 TI - Renal complications in a patient with A-to-G mutation of mitochondrial DNA at the 3243 position of leucine tRNA. AB - A 27-year-old woman with short stature, sensorineural deafness, and renal dysfunction was hospitalized for evaluation. The serum lactate and pyruvate concentrations were elevated. The analysis of her mitochondrial DNA revealed an A to-G mutation of the tRNA(Leu (UUR)) gene at the 3243 position. Renal biopsy revealed many sclerotic glomeruli, advanced tubulointerstitial changes, and numerous swollen mitochondria of the tubular cells. It was concluded that the patient's mitochondrial gene mutation was etiologically related to her nephropathy. The clinicopathologic features of this patient, as contrasted to the previous reports, suggested that renal involvement due to this mitochondrial gene mutation can be heterogeneous. PMID- 11868598 TI - Water intake and 24-hour blood pressure monitoring in a patient with nephrogenic diabetes insipidus caused by a novel mutation of the vasopressin V2R gene. AB - A 47-year-old man presented with polydipsia, which had had since childhood, and recent onset of hypertension. Genetic analysis proved that he had nephrogenic diabetes insipidus caused by a novel mutation (deletion of 6 amino acids between G107 and C112) in the vasopressin V2 receptor gene. Results of 24-hour blood pressure monitoring disclosed a greater dipping pattern and greater blood pressure variability during waking hours than in male patients with only essential hypertension. This characteristic blood pressure profile may result from daily occurrence of free water depletion, as further observation indicated that water deprivation was associated with a reduction in blood pressure. PMID- 11868599 TI - Two cases of Sjogren's syndrome with multiple bullae. AB - Here, we report two rare female cases of Sjogren's syndrome with multiple bullae, involving a 66- and a 51-year-old. Neither had any obvious pulmonary complaint. Chest radiographs and high-resolution CT (HRCT) scans showed interstitial linear and nodular opacities and multiple bullae. In the first case spirometry indicated an obstructive change judged by FEV1.0 and V50/V25. In both cases, histologic examination of the lung revealed thickening of alveolar septa and interstitial mononuclear cell infiltration. In the first case the bullae decreased in size with corticosteroid treatment. Airway narrowing due to peribronchiolar mononuclear cell infiltration causes a check-valve mechanism, which may lead to bullae formation. Although a rare occurrence, it is important to recognize that cystic or bullous lung disease can accompany Sjogren's syndrome. PMID- 11868600 TI - Increased serum thymidine kinase activity in acute sarcoidosis. AB - This is the first case report of acute sarcoidosis with increased serum thymidine kinase (TK) activity. A 43-year-old male presented fever, swelling of parotid glands, lymphadenopathy, and peripheral neuropathy. Sarcoidosis was pathologically diagnosed by lung and parotid gland biopsy. His serum TK, which was increased to 11.2 U/l at diagnosis (normal <5 U/l), normalized after glucocorticoid therapy. Serum TK has been considered as a good marker of the proliferative activity of various types of neoplasms. Its rise in sarcoidosis has, however, not been described. Because acute sarcoidosis sometimes resembles malignant lymphoma, the possible rise of serum TK in sarcoidosis may be worthy of note. PMID- 11868601 TI - Rapidly expanding lung abscess caused by Legionella pneumophila in immunocompromised patients: a report of two cases. AB - We describe two cases of lung abscess caused by Legionella pneumophila in immunocompromised patients. The first case had been treated initially with 60 mg prednisolone for ulcerative colitis, and L. pneumophila serogroup 1 was isolated from sputum samples after cavitation of the lung lesion. The second case was diagnosed as plasma cell lymphoma at post-mortem examination. L. pneumophila serogroup 5 was isolated from the contents of lung abscess, together with Enterococcus faecium and Prevotella intermedia in the post-mortem examination. Lung abscess caused by Legionella is unusual. Here, we discuss the difficulty of diagnosis of legionellosis in patients with unusual chest radiographic findings. PMID- 11868602 TI - Smoldering myeloma associated with zonisamide treatment. AB - A 39-year-old man was found to have hyperproteinemia after being treated with zonisamide for 10 years. Laboratory examination revealed a serum M-protein which consisted of IgG (lambda) and an increased number of plasma cells in the bone marrow, resulting in a diagnosis of smoldering myeloma. Considering his age, zonisamide was suspected to play an etiologic role in the occurrence of smoldering myeloma. Zonisamide was changed to sodium valproate. Subsequently the M-protein did not increase over 13 months. When zonisamide is used, the monitoring of serum levels of M-protein and patterns of gammaglobulin is warranted. PMID- 11868604 TI - Rippling muscle syndrome preceding malignant lymphoma. AB - A 46-year-old woman presented with rippling muscle phenomenon. She complained of uncomfortable muscular stiffness of extremities and abdominal wall. Muscle contraction was easily elicited by percussion, which was visible from the surface and propagated in a rolling manner. The mounding (or myoedema) phenomenon was also remarkable. Three years later, malignant lymphoma (histologically, lymphoplasmacytoid lymphoma) was found in the sacrum. The lymphoma subsided with treatment by vincristine, cyclophosphamide, doxorubicin and prednisolone. Serum IgG as well as creatine kinase values were normalized. The rippling phenomenon also responded to the treatment. The present rippling muscle syndrome might be of a paraneoplastic or autoimmune origin related to lymphoma, although the evidence seemed indirect. We discussed the role of the internal membrane system of the skeletal muscle in the pathogenesis of rippling muscle. PMID- 11868603 TI - Motor dominant neuropathy in Sjogren's syndrome: report of two cases. AB - Most of the peripheral neuropathies in Sjogren's syndrome (SS) are sensory- or autonomic-dominant. In this report, we present two cases of a rare type of neuropathy, motor dominant neuropathy, in SS. One showed signs similar to those of Guillain-Barre syndrome, and the other showed signs characteristic of chronic inflammatory demyelinating polyradiculoneuropathy. These patients received i.v. immunoglobulin therapy. To our knowledge, this is the first report indicating that i.v. immunoglobulin has beneficial effects on motor dominant neuropathy in SS. PMID- 11868606 TI - Systemic lupus erythematosus related transverse myelitis presenting longitudinal involvement of the spinal cord. AB - Lupus-related transverse myelitis is a rare but serious complication. A 25-year old Japanese woman with systemic lupus erythematosus (SLE) was admitted because of numbness of the face and left upper extremity, headache, and intermittent fever. Six days later, she developed tetraplegia. MRI of the spinal cord showed longitudinal high intensity signals from medulla oblongata to C5, and from Th12 to conus medullaris on T2-weighted image. These MRI findings were consistent with acute catastrophic neurological abnormalities. Despite administration of the combination of methylprednisolone and cyclophosphamide pulse therapies, as well as plasmapheresis, her condition did not improve. Any vasculopathy in addition to the autoimmune pathogenesis, and narrow therapeutic window may relate to the present refractory case. PMID- 11868605 TI - Systemic lupus erythematosus complicated by cytomegalovirus-induced hemophagocytic syndrome and colitis. AB - Here, we report a case of systemic lupus erythematosus (SLE) complicated by cytomegalovirus (CMV)-induced hemophagocytic syndrome (HPS) and colitis. A 44 year-old woman with SLE was treated with corticosteroid and cyclophosphamide for lupus nephritis. Although her lupus nephritis improved, fever, progressive pancytopenia and intestinal bleeding were observed. A bone marrow aspiration showed an increase in mature histiocytes with hemophagocytosis. In addition, a colonoscopy showed hemorrhagic colitis with ulcer and the biopsy specimen from the colon revealed typical CMV cells with CMV inclusions confirmed by immunohistochemistry. Furthermore, a large number of CMV antigen-positive leukocytes was detected, suggesting an active CMV infection. CMV infection is serious in compromised hosts. Therefore clinicians should be aware of the clinical settings in which this infection can arise and the target organs potentially affected in order to initiate the appropriate intervention. PMID- 11868607 TI - Systemic lupus erythematosus associated with massive ascites and pleural effusion in a patient who presented with disseminated intravascular coagulation. AB - A case of systemic lupus erythematosus (SLE) associated with serositis presenting with disseminated intravascular coagulation (DIC) is reported. A 53-year-old woman was admitted because of a fever. Laboratory tests revealed increased plasma levels of fibrinogen degradation products (FDP) and FDP-D-dimer, high titers of anti-nuclear antibody, high serum levels of anti-DNA antibody, immune complexes, decreased serum complements, and persistent proteinuria. A CT scan showed massive ascites and pleural effusion, marked edema and swelling of the mesenterium. The patient's condition and immunological abnormalities improved after steroid therapy. The association of DIC and lupus serositis has never been described in the literature. PMID- 11868608 TI - Pictures in clinical medicine. Fungal endophthalmitis and Churg-Strauss syndrome. PMID- 11868609 TI - Pulmonary involvement in Sjogren's syndrome. PMID- 11868610 TI - Sjogren's syndrome. Yes autoreactive lymphocytes, why? Virus or gene? PMID- 11868611 TI - Systemic lupus erythematosus complicated by cytomegalovirus-induced hemophagocytic syndrome and colitis. PMID- 11868612 TI - Involvement of hypoxia-inducible factor 1 in human cancer. AB - Hypoxia-inducible factor 1 (HIF-1) mediates transcriptional responses to hypoxia. HIF-1 is composed of an O2- and growth factor-regulated HIF-1alpha subunit and a constitutively-expressed HIF-1beta subunit. Four lines of evidence indicate that HIF-1 contributes to tumor progression. First, HIF-1 controls the expression of gene products that stimulate angiogenesis, such as vascular endothelial growth factor, and promote metabolic adaptation to hypoxia, such as glucose transporters and glycolytic enzymes, thus providing a molecular basis for involvement of HIF-1 in tumor growth and angiogenesis. Second, in mouse xenograft models, tumor growth and angiogenesis are inhibited by loss of HIF-1 activity and stimulated by HIF 1alpha overexpression. Third, immunohistochemical analyses of human tumor biopsies indicate that HIF-1alpha is overexpressed in common cancers and that the level of expression is correlated with tumor grade, angiogenesis, and mortality. Fourth, in addition to intratumoral hypoxia, genetic alterations in tumor suppressor genes and oncogenes induce HIF-1 activity. PMID- 11868613 TI - Adenosine triphosphate-sensitive potassium (K(ATP)) channel activity is coupled with insulin resistance in obesity and type 2 diabetes mellitus. AB - ATP sensitive potassium (K(ATP)) channels reside in the plasma membrane of many excitable cells such as pancreatic beta-cells, heart, skeletal muscle and brain, where they link cellular metabolic energy to membrane electrical activity. They are composed of two subunits, K+ ion selective pore (Kir) and sulfonylurea receptor (SUR). In addition to the central role of pancreatic beta-cell K(ATP) channels in glucose-mediated insulin secretion, several lines of evidence support the hypothesis that K(ATP) channels modulate glucose transport in the insulin target tissues. Inhibition of K(ATP) channels by glibenclamide or gliclazide or an increase in intracellular ATP during hyperglycemia (glucose effect) or exercise facilitates glucose utilization, while activation of the channels by potassium channel openers, hypothermia (cardiac surgery), or ischemic damage (myocardial and brain infarction) reduces glucose uptake induced by insulin or hyperglycemia. Because insulin action has been known to depend on the energy level of the target cells, K(ATP) channel may function as an effector in this respect. It is now evident that long chain acyl-CoA esters, metabolically active forms of fatty acids, are the most potent and physiologically important activator of K(ATP) channels. Thus, I suppose that the sustained activation of K(ATP) channels by long chain fatty acyl-CoA seems to be a missing link between lipotoxicity and insulin resistance in obesity and type 2 diabetes mellitus. PMID- 11868614 TI - Prognosis of economy class syndrome treated in intensive care unit. AB - OBJECTIVE: The aim of this study was to clarify the prognosis of economy class syndrome (pulmonary thromboembolism following long distance flight) patients discharged from the intensive care unit. METHOD: Medications, daily activities, recurrence of pulmonary thromboembolism, and quality of life were investigated for an average of 471 days following discharge from the intensive care unit. The information from the patients was obtained by means of a mailed questionnaire or telephone interview. PATIENTS: Ten patients with economy class syndrome were enrolled in this study. Nine were female and one was male (61+/-9 years). RESULTS: In addition to appropriate acute care, additional management was given including the placement of an inferior vena cava filter in 3 patients and a course of oral anticoagulation therapy in 7 patients. None of them died or had recurrence of pulmonary thromboembolism. Four patients took subsequent air flights. No thromboembolic episodes were seen during the follow-up period. CONCLUSION: The long-term prognosis in these patients can be favorable with appropriate management of the acute episode in conjunction with adequate preventive measures. PMID- 11868615 TI - Reasons for the delays in the definitive diagnosis of lung cancer for more than one year from the recognition of abnormal chest shadows. AB - OBJECTIVE: Primary lung cancer generally has a poor prognosis if not diagnosed at an early stage. But some lung cancers grow very slowly. In particular, adenocarcinoma is sometimes observed for years with no change of tumor size. In this study, we examined the reasons for the delays in reaching a definitive diagnosis of lung cancer. METHODS: We retrospectively reviewed primary lung cancer cases between January 1995 and December 1999 and examined those whose definitive diagnoses were delayed for more than a year. RESULTS: A total of 222 primary lung cancers were diagnosed. Of those, 19 patients (group A, 8.6%) were diagnosed after more than a year, and the other 203 (group B, 91.4%) were diagnosed within one year. The proportion of women in group A was significantly higher than that in group B (p<0.05). The mean age of group A was significantly younger than that of group B (p<0.05). The Brinkman Index of group A was significantly lower than that of group B (p<0.05). The histologic types were significantly different between the two groups (p<0.05). In group A, 18 patients (94.7%) had adenocarcinomas. Five primary reasons for the delays in group A were identified: 1) Four patients were tentatively diagnosed as inflammation or benign tumor on CT and were consequently not followed-up. 2) The chest CT shadows in 6 patients were suspected lung cancers but transbronchial lung biopsy findings did not show malignancy. 3) Four patients were tentatively diagnosed as inflammation or benign tumor on CT, but the tumors showed only very slow growth or no change at all. 4) The chest CT shadows of 2 patients were suspected lung cancer, but the patients refused to undergo video-assisted thoracic surgery (VATS) or closer examination. 5) Three patients did not consult medical facilities for a second examination. CONCLUSIONS: Many of the adenocarcinomas reviewed in our study grew slowly or remained unchanged for years. Doctors are mainly responsible for the delays in the definitive diagnosis and should aggressively perform VATS or closer examinations without hesitation. PMID- 11868616 TI - Beyond pretrial publicity: legal and ethical issues associated with change of venue surveys. AB - To combat the effects of pretrial publicity, a court may consider a change of venue, moving the trial to a jurisdiction that has been exposed to little publicity about the case. Social scientists can be of valuable assistance in measuring community attitudes regarding defendant level of guilt and case familiarity. This paper discusses obstacles faced by the authors when conducting change of venue surveys and presenting their findings to the court, including challenges to their ability to act and be viewed as scientists, and pressures to do that which contradicts empirical research in order to please the court. Future research should examine factors predicting judges' decisions on change of venue motions and issues associated with small venues and civil cases. PMID- 11868617 TI - Afterword: the past, present, and future of applied pretrial publicity research. PMID- 11868618 TI - Studying pretrial publicity effects: new methods for improving ecological validity and testing external validity. AB - Although research examining the effects of pretrial publicity (PTP) on individuals' appraisals of a defendant and verdict decision making generally has been found to be internally valid, the external validity has been questioned by some social scientists as well as lawyers and judges. It is often proposed that the verisimilitude (or ecological validity) ofthe research should be increased in the service of increasing external validity; however, increasing verisimilitude can be costly in terms of both time and money. It is proposed that the Internet is a viable means of conducting PTP research that allows high verisimilitude without high costs. This is demonstrated with a study in which we used the Internet to examine PTP effects in an actual trial as it was taking place. Successful use of the Internet to conduct experimental research in other areas of psychology and law is discussed, as well as the importance of future research examining whether independent variables interact with methods in ways that undermine the generalizability of research findings. PMID- 11868619 TI - Pretrial publicity and civil cases: a two-way street? AB - Published pretrial publicity (PTP) research has been conducted almost exclusively with criminal cases and has focused on PTP that is detrimental to the defense. The current research examined the effects of PTP in a civil case to determine if PTP can have a biasing effect against either the defendant or the plaintiff in civil litigation. In Experiment 1, participants exposed to PTP biased against the defendant were more likely to reach a liable verdict than participants who read a control article or PTP biased against the plaintiff Experiment 2 demonstrated that a judicial admonition did not reduce the biasing effect of PTP about a civil defendant. However, participants given the admonition both before and after the trial evidence viewed the defendant as less culpable than participants given the admonition after the trial only or not at all. The implications for the legal system are discussed. PMID- 11868620 TI - The effects of general pretrial publicity on juror decisions: an examination of moderators and mediating mechanisms. AB - Two studies examined three moderators (gender, attitudes, and media slant) and four mediators (accessibility, evidence importance, evidence plausibility, and standards of guilt) of general pretrial publicity's influence on juror decisions. In Study 1, participants who watched a prodefense rape story were more likely to report that they would need more inculpatory evidence to convict a defendant of rape than were participants who watched a proprosecution rape story. In Study 2, participants watched news stories, one of which was a proprosecution rape story, a prodefense rape story, or a nonrape story. In an ostensibly unrelated study, participants indicated their attitudes toward rape, watched a rape trial, and provided trial and witness ratings. Accessibility did not mediate the media effects on participants' judgments of rape importance; however, attitudes moderated media effects. Rape news influenced juror ratings of the importance of evidence about the complainant's behavior. Finally, media altered the standards participants used to determine defendant guilt. Implications for understanding the mechanisms responsible for pretrial publicity effects are discussed. PMID- 11868621 TI - Case studies of pre- and midtrial prejudice in criminal and civil litigation. AB - This paper presents a number of case studies involving pre- and midtrial prejudice in criminal and civil litigation. The cases reveal deficiencies in the way that prejudicial publicity has been conceptualized and operationalized in many simulation experiments. The studies reveal that potential juror prejudices that concern lawyers and judges involve more than just main effects of mass media. Pre- and midtrial prejudice also involves more general prejudices, gossip and rumor, the assertion of community normative values about justice, and conformity pressures. Four categories of prejudice recognized in American law are described and labeled: interest, specific, generic, and conformity prejudice. The case studies also reveal interesting dynamics involving "minimization" and inconsistency in jurors' self-reports of attitudes which contradict a commonly held judicial view that superficial questioning is sufficient to uncover prejudice. Despite identifying deficiencies in the simulation literature, the paper concludes that experimental research is necessary to provide answers to causal questions that case studies and field research cannot ordinarily provide. PMID- 11868622 TI - Aggressive angiomyxoma of the scrotum. AB - Aggressive angiomyxoma is a rare and nonmetastasizing soft tissue tumor of the pelvis and perineum and occurs almost exclusively in adult females. It infiltrates locally and has a high risk of local recurrence. Recommended treatment of the symptomatic patient is wide excision with tumor-free margins and close postoperative monitoring. Herein, a case of aggressive angiomyxoma in an adult male is described, which arose in the scrotum over 12 months. The tumor showed an intermediate signal on T1-weighted MRI images. Contrast-enhanced T1 weighted images showed good enhancement. Wide excision of the tumor was performed. The surgical specimen measured 7 x 5 x 5 cm in size and weighed 80 g. The tumor's surface was smooth and had a gelatinous cut surface. Grossly, it was encapsulated with a pleura-like membrane and had a finger-like projection. Microscopically, sections showed many walled vessels of various sizes, collagen fibrils, a loose myxoid background. and spindle stroma cells. MRI and CT showed the angiomatous and myxomatous nature of the tumor wall. To the authors' knowledge, this is the first report to describe MRI findings in scrotal angiomyxoma. PMID- 11868623 TI - Interleukin-1 beta and tumor necrosis factor-alpha in normal/infertile men. AB - Cytokines play an important role in intercellular communications. Human sperm contains a wide spectrum of cytokines. such as interleukin-1 beta (IL-1beta) and tumor necrosis factor alpha (TNF-alpha). Their effects on semen quality are subject to debate. The aim of this study was to determine concentrations of IL 1beta and TNF-alpha in normal fertile men and in different groups of male infertility in an attempt to clarify the physiology and suggest possible clinical uses. Sixty-six subfertile male patients with varicocele (n = 22). infection of accessory genital glands (n = 14), varicocele plus infection (n = 4), chronic epididymitis (n = 8). post-renal transplantation status (n = 5), idiopathic oligoasthenoteratospermia (n = 9), cryptorchidism (n = 1), and homozygous beta thalassemia (n = 3) as well as 5 male controls were studied through history, physical examination, spermiograms, plasma basal hormonal levels, and IL-1beta and TNF-alpha levels in seminal fluid. There was no significant statistical difference regarding IL-1beta and TNF-alpha among fertile men and subfertile patients of any cause. 1L-1beta and TNF-alpha were in tight positive correlation (p<.001). Determination of IL-1beta and TNF-alpha does not provide useful information in male routine infertility workup. Nevertheless, a better understanding of these mediators in semen of normal men and infertile patients may contribute to a new approach to the management of male infertility. PMID- 11868624 TI - Effects of gonadal hormones on thymus gland after bilateral ovariectomy and orchidectomy in rats. AB - This study examined the histological changes that occurred in the thymus gland after gonadectomy and the administration of various sex steroids following gonadectomy. Male and female Wistar albino rats that were 6 weeks of age were used. The rats were subjected to bilaterally gonadectomy and then gonadal steroid hormones (testosterone, estrogen, and progesterone, 2.5 mg/kg) were given. Effects of gonadal steroid hormones on the thymus gland were microscopically examined. Thymic weight increased in all the groups after gonadectomy. Testosterone, estrogen, and estrogen + progesterone treatment decreased thymic weight after gonadectomy. Progesterone treatment also decreased weight, but there was no statistical significance. In the light microscopy, testosterone and estrogen treatment induced a loss of lymphoid elements from the thymic cortex, increased the number of phagocytic macrophages and mast cells, and enlarged blood vessels and connective tissue were observed in the thymic medulla. In the electron microscopic study it was observed that rough endoplasmic reticulum enlarged in the thymic lymphocytes. The same results were also found after estrogen + progesterone treatment. No histologically identifiable changes were observed in the thymus gland after progesterone treatment. This study demonstrated that the thymus gland undergoes involution after testosterone and estrogen treatment, but not progesterone, following gonadectomy. PMID- 11868625 TI - Shedding of immature germ cells. AB - The immature germ cells (IGC) constitute the highest percentage (90%) of nonsperm cells (NSpC) in ejaculates from fertile or infertile men. The objective of this study was to evaluate IGC concentration and the IGC/(IGC + Sp) ratio, in normozoospermia and dispermia. Normozoospermia from men with proven fertility (NPF). nonproven fertility (NNPF). dispermia (D) and semen samples with excessive shedding of immature germ cells (GI 1.7 x 10(6) to 5 x 10(6) IGC/mL and GII > 5.0 x 10(6) IGC/mL) were used in this study. The mean value +2 SD for the NNPF (1.7 x 10(6)/mL) and the value proposed by WHO (5 x 10(6)/mL) were employed to define GI and GII groups. IGC concentration is statistically different in the studied groups. The IGC/Sp ratio showed a significant difference only between the NNPF and the D. When comparing semen parameters (Sp/ejaculate. grade (a) motility and morphology) there was a highly significant difference between NNPF and GI and GII: no difference was found between GI and GII. While studying 200 cases of dispermias 83% showed a high shedding of immature germ cells. The cytological study of nonsperm cells and the count and identification of the immature germ cells could be used to evaluate the dispermic disorders. PMID- 11868627 TI - Comparison of centrifugation- and noncentrifugation-based techniques for recovery of motile human sperm in assisted reproduction. AB - To compare standard density gradient centrifugation sperm preparation with a novel non-centrifugation-based dual-chamber capillary dish in efficiency for motile human sperm separation, approximately 3 mL fresh ejaculate specimens was obtained from 21 men (median age = 32 years. range 26-42 years) undergoing infertility evaluation. For each specimen, half of the sample was processed with a standard 45%/90% density gradient preparation (PureSperm. Nidacon International, Gothenburg, Sweden) followed by semen analysis. The other half was incubated in the Zech glass capillary dish (Astromedtec, Salzburg, Austria) consisting of 2 concentric wells overlaid by a U-ring and coverglass. After approximately 3 h, a 1-mL sample was taken from the central chamber and analyzed. Percentage motile sperm recovery, absolute (motile) cell number, and path velocities were compared for spermatozoa obtained from both methods. Both techniques reduced overall sperm concentration while enriching specimens with more motile spermatozoa. A trend towards higher % recovery of motile spermatozoa (p = .264) was observed with the Zech device, but at a cost of fewer absolute numbers of higher velocity cells (p = .004). The Zech device, therefore, localized a very small population of motile sperm without exposure to centrifugation stress, which has been considered potentially harmful to spermatozoa. This technique could theoretically improve efficiency by reducing time required to identify motile cells in in vitro fertilization where intracytoplasmic sperm injection is planned. However, refinements in incubation interval and suspension volumes are needed before this technique can be considered comparable to the density gradient method in recovering sperm for use in intrauterine insemination. PMID- 11868626 TI - Acrosome reaction in fertile and subfertile boar sperm. AB - The main purpose of sperm evaluation is to predict its fertilizing ability. However, basic sperm test results show a low correlation with fertilizing ability. The purpose of this study was to determine whether there is an association between acrosome reaction (AR) and the incidence of subfertility of normal sperm boar. The production records of 22 farms were analyzed to identify boars with low fertility and/or prolificity, classified as subfertile. Twenty-two subfertile boar semen samples were analyzed and compared with 51 samples of fertile boars. Sperm were capacitated during 4 h at 39 degrees C. viability was determined by bisbenzimide (Hoechst-33258) staining. Acrosome reaction was assessed with fluorescein isothiocyanate conjugated Pisum sativum agglutinin. The percentage of spontaneous acrosome reaction (SAR) was not significantly different in fertile (4.5%) and subfertile boars (4.75%) (p > .05). Nevertheless, the percentage of progesterone-induced acrosome reaction (IAR) was significantly lower in subfertile boars (5.75%) as compared with fertile boars (10%) (p < .01). These results suggest that assessment of IAR in vitro may be a useful parameter to identify subfertility in boars. PMID- 11868628 TI - Heparin binding sites are present at a higher concentration on sperm of subfertile men than donors of known fertility. AB - Heparin sulfate has been shown to both stimulate sperm capacitation and cause decondensation of sperm chromatin. Recent studies have shown that sperm chromatin decondensation following exposure to a low concentration of heparin sulfate is inversely correlated with penetration ability. This study shows that sperm from subfertile patients contain a higher (p < .05) concentration of heparin binding sites than sperm from donors of known fertility. No differences were observed in the binding affinity of 3H-labeled heparin to donor and patient sperm. Binding was specific for sulfated heparin and dextran. FITC-labeled heparin bound to a higher percentage (p < .01) of patient sperm (39.6 +/- 3.1) than donor sperm (12.3 +/- 1.7). A relationship is demonstrated between heparin binding sites and sperm chromatin decondensation following heparin cxposure, both of which are increased in sperm from subfertile men. PMID- 11868629 TI - Treatment of sperm with subnormal host scores with chymotrypsin/viable pregnancy after IUI. AB - Men with low hypoosmotic swelling test (HOST) scores (<50%) rarely achieve a pregnancy with intercourse or conventional intrauterine insemination (IUI) or even IVF. The defect seems to be related to a toxic factor attached to the sperm that can be transferred to the zona pellucida, which ultimately interferes with implantation. A small case series showed optimistic pregnancy outcome with treatment of the sperm with low HOST scores with chymotrypsin. However, the live pregnancy rate in 90 subsequent IUI cycles in men whose low HOST scores were improved by chymotrypsin was only 3.3%. IVF with ICSI remains the only highly effective treatment of this disorder. PMID- 11868630 TI - Expression of lectin receptors on the membrane surface of sperm of fertile and subfertile boars by flow cytometry. AB - Studies suggest that carbohydrates are important in different stages of fertilization. Plasma membrane changes accompanying in vitro capacitation and acrosome reaction (AR), such as removal or appearance of specific glycoproteins, have been studied using lectins that bind specifically to carbohydrate residues. In specialized artificial insemination farms and semen production centers, identification of boars with decreased fertilization ability (subfertility) is a newborn necessity. This investigation is a sequential study to determine the kinetics of surface carbohydrates turnover during in vitro capacitation and AR in fertile and subfertile boar sperm. Flow cytometry determinations of the binding of three FITC-labeled lectins were assessed. WGA binding was significantly lower in fresh, capacitated, and acrosome-reacted sperm of subfertile boars than in fertile boars. Con-A binding was not significantly different in fresh sperm of fertile and subfertile boars. However. Con-A labeling in capacitated, and acrosome-reacted sperm differed significantly in both groups. UEA binding increased only in capacitated sperm of subfertile boars. These findings could be used as indicators of capacitation and AR and may also be a good indicator of sperm fertilizing ability in boars. PMID- 11868631 TI - Procarbazine effects on spermatogenesis in golden hamster: a flow cytometric evaluation. AB - The response of hamster testis to the administration of 450mg/kg procarbazine (PCB) over a period of 4 weeks was evaluated. Flow cytometry was used to investigate changes in cell populations in testicular single cell suspensions and to correlate these changes with those observed in histological sections. PCB caused significant decrease in testicular and epididymal weight and a drastic reduction in haploid cells and spermatogenic arrest, demonstrating variation among the test animals. The results obtained confirm previous observations concerning detrimental effects of PCB upon spermatogenesis in species such as the rat and mouse, though its effect on hamster testis is milder and does not include the germinal stem cells. The histological evaluation of the testis showed a good correlation with flow cytometric evaluation, emphasizing the usefulness of this method in providing quantitative and rapid results. PMID- 11868632 TI - Rehabilitating Afghanistan. PMID- 11868633 TI - Addressing the challenges of queues. PMID- 11868634 TI - Oral corticosteroids for poison ivy dermatitis. PMID- 11868635 TI - Preconceptional sex selection. PMID- 11868636 TI - Mandatory vaccination of health care workers. PMID- 11868637 TI - Cardiovascular effects of strenuous exercise in adult recreational hockey: the Hockey Heart Study. AB - BACKGROUND: More than 500,000 men play "gentlemen's" recreational hockey in Canada, but the safety of this exercise has not been studied. Exercising at extremes of intensity has been associated with an increased risk of cardiac events. Our objective was therefore to determine baseline cardiac risk factors among adult recreational hockey players and to measure any cardiac abnormalities they experienced while playing hockey. METHODS: We assessed baseline cardiac risk factors in 113 male volunteers recruited from a recreational hockey league. Each subject underwent holter electrocardiographic monitoring before, during and after at least one hockey game (maximum of 115 holter data sets). We used the data to assess exercise heart rate, arrhythmias and ST-segment changes and for correlation with symptoms and other predictors of fitness. RESULTS: For all participants, maximum heart rate (HRmax) (mean 184 [standard deviation 11] beats/min) was greater than target exercise heart rate (calculated as 55% to 85% of age-predicted HRmax), and in 87 (75.6%) of the 115 holter data sets, the heart rate exceeded the age-predicted HRmax. The mean period for which heart rate exceeded 85% of the age-predicted HRmax was 30 (SD 13) min. For 80 (70.1%) of 114 data sets, heart rate recovery was poor. Nonsustained ventricular tachycardia was seen in data from 2 holter monitoring sessions and ST-segment depression in data from 15 sessions. INTERPRETATION: The physical activity pattern that occurred during recreational hockey caused cardiac responses that might be dangerous to players' health. More specifically, the players exceeded target and maximum heart rates, had poor heart rate recovery after exercise, and had episodes of nonsustained ventricular tachycardia and ST-segment depression of uncertain clinical significance. PMID- 11868638 TI - Transfusion practices for elective orthopedic surgery. AB - BACKGROUND: Use of blood conservation techniques in elective surgery reduces the risk of infection and transfusion reactions that result from using allogeneic blood products. We examined the transfusion practice and blood conservation strategies for elective orthopedic procedures in 19 Canadian hospitals. METHODS: We reviewed the medical records of patients who underwent total hip or knee joint arthroplasty between June 1998 and January 1999 in a convenience sample of 19 hospitals to determine the pre- and postoperative hemoglobin concentrations, concurrent medical conditions, participation status in an autologous blood donation program, use of other blood conservation techniques, and occurrence of allogeneic and autologous transfusions. Patients were considered eligible for autologous blood donation if they weighed at least 25 kg, were in good general health without major cardiac conditions and had a hemoglobin concentration of at least 110 g/L. RESULTS: We reviewed 4535 medical records. Of the 4422 patients whose eligibility status was known, 2561 (57.9%) were eligible to participate in an autologous blood donation program. Only 842 (18.6%) of the patients predonated blood. Patients who did not predonate blood were older (mean age 70.1 v. 63.8 years) and were more likely to have concomitant medical conditions (60.3% v. 37.9%) than those who did predonate. Overall, 30.6% (95% confidence interval [CI] 29.1%-32.1%) of the patients who did not predonate blood received allogeneic transfusions. For patients who predonated, the rate of allogeneic transfusion was 14.1% (95% CI 11.8%-16.5%). The frequency with which blood conservation techniques other than autologous blood donation were used was minimal (in 2.4% of all cases). INTERPRETATION: The use of blood conservation techniques among hospitals in Canada remains low. Only a minority of eligible patients participated in an autologous blood donation program. PMID- 11868639 TI - Outcomes of planned home births versus planned hospital births after regulation of midwifery in British Columbia. AB - BACKGROUND: The choice to give birth at home with a regulated midwife in attendance became available to expectant women in British Columbia in 1998. The purpose of this study was to evaluate the safety of home birth by comparing perinatal outcomes for planned home births attended by regulated midwives with those for planned hospital births. METHODS: We compared the outcomes of 862 planned home births attended by midwives with those of planned hospital births attended by either midwives (n = 571) or physicians (n = 743). Comparison subjects who were similar in their obstetric risk status were selected from hospitals in which the midwives who were conducting the home births had hospital privileges. Our study population included all home births that occurred between Jan. 1, 1998, and Dec. 31, 1999. RESULTS: Women who gave birth at home attended by a midwife had fewer procedures during labour compared with women who gave birth in hospital attended by a physician. After adjustment for maternal age, lone parent status, income quintile, use of any versus no substances and parity, women in the home birth group were less likely to have epidural analgesia (odds ratio 0.20, 95% confidence interval [CI] 0.14-0.27), be induced, have their labours augmented with oxytocin or prostaglandins, or have an episiotomy. Comparison of home births with hospital births attended by a midwife showed very similar and equally significant differences. The adjusted odds ratio for cesarean section in the home birth group compared with physician-attended hospital births was 0.3 (95% CI 0.22-0.43). Rates of perinatal mortality, 5-minute Apgar scores, meconium aspiration syndrome or need for transfer to a different hospital for specialized newborn care were very similar for the home birth group and for births in hospital attended by a physician. The adjusted odds ratio for Apgar scores lower than 7 at 5 minutes in the home birth group compared with physician attended hospital births was 0.84 (95% CI 0.32-2.19). INTERPRETATION: There was no increased maternal or neonatal risk associated with planned home birth under the care of a regulated midwife. The rates of some adverse outcomes were too low for us to draw statistical comparisons, and ongoing evaluation of home birth is warranted. PMID- 11868640 TI - Anal incontinence after childbirth. AB - BACKGROUND: Incontinence of stool and flatus are frequent complications of childbirth. We examined the prevalence and possible causes of these adverse outcomes in a large cohort of women. METHODS: We studied 949 pregnant women who gave birth in 5 hospitals in 1995/96 in the province of Quebec. These women, participants in a randomized controlled trial of prenatal perineal massage, completed a self-administered questionnaire 3 months after giving birth. RESULTS: Three months after delivery 29 women (3.1%) reported incontinence of stool, and 242 (25.5%) had involuntary escape of flatus. Incontinence of stool was more frequent among women who delivered vaginally and had third- or fourth-degree perineal tears than among those who delivered vaginally and had no anal sphincter tears (7.8% v. 2.9%). Forceps delivery (adjusted risk ratio [RR] 1.45, 95% confidence interval [CI] 1.01-2.08) and anal sphincter tears (adjusted RR 2.09, 95% CI 1.40-3.13) were independent risk factors for incontinence of flatus or stool or both. Anal sphincter injury was strongly and independently associated with first vaginal birth (RR 39.2, 95% CI 5.4-282.5), median episiotomy (adjusted RR 9.6, 95% CI 3.2-28.5), forceps delivery (adjusted RR 12.3, 95% CI 3.0-50.4) and vacuum-assisted delivery (adjusted RR 7.4, 95% CI 1.9-28.5) but not with birth weight (adjusted RR for nirth weight 4000 g or more: 1.4, 95% CI 0.6-3.0) or length of the second stage of labour (adjusted RR for second stage 1.5 hours or longer compared with less than 0.5 hours: 1.2, 95% CI 0.5-2.7). INTERPRETATION: Anal incontinence is associated with forceps delivery and anal sphincter laceration. Anal sphincter laceration is strongly predicted by first vaginal birth, median episiotomy, and forceps or vacuum delivery but not by birth weight or length of the second stage of labour. PMID- 11868641 TI - The double-edged blade of recreational hockey. PMID- 11868642 TI - Reducing the risk of allogeneic blood transfusion. PMID- 11868643 TI - Are home births safe? PMID- 11868644 TI - Cesarean section versus forceps-assisted vaginal birth: it's time to include pelvic injury in the risk-benefit equation. PMID- 11868645 TI - Bioethics for clinicians: 28. Protestant bioethics. AB - "Protestant" is a term applied to many different Christian denominations, with a wide range of beliefs, who trace their common origin to the Reformation of the 16th century. Protestant ideas have profoundly influenced modern bioethics, and most Protestants would see mainstream bioethics as compatible with their personal beliefs. This makes it difficult to define a uniquely Protestant approach to bioethics. In this article we provide an overview of common Protestant beliefs and highlight concepts that have emerged from Protestant denominations that are particularly relevant to bioethics. These include the sovereignty of God, the value of autonomy and the idea of medicine as a calling as well as a profession. Most Canadian physicians will find that they share certain values and beliefs with the majority of their Protestant patients. Physicians should be particularly sensitive to their Protestant patients' beliefs when dealing with end-of-life issues, concerns about consent and refusal of care, and beginning-of-life issues such as abortion, genetic testing and the use of assisted reproductive technologies. Physicians should also recognize that members of certain Protestant groups and denominations may have unique wishes concerning treatment. Understanding how to elicit these wishes and respond appropriately will allow physicians to enhance patient care and minimize conflict. PMID- 11868646 TI - Nephrology: 2. Evaluation of asymptomatic hematuria and proteinuria in adult primary care. PMID- 11868647 TI - Hospitals and the environment. PMID- 11868648 TI - Cardiovascular disease and risk in the Aboriginal population. PMID- 11868649 TI - Investment in international health will pay massive dividend: WHO. PMID- 11868650 TI - How did the situation at the Queensway-Carleton get so nasty? PMID- 11868651 TI - Auditor general targets Health Canada spending. PMID- 11868652 TI - In wake of 9/11, drug-patent rules change. PMID- 11868653 TI - Popularity of alternative health care providers continues to grow. PMID- 11868654 TI - More provinces protecting MDs who practise alternative medicine. PMID- 11868655 TI - Evolutionary innovations overcome ancestral constraints: a re-examination of character evolution in male sepsid flies (Diptera: Sepsidae). PMID- 11868656 TI - Structure and metamorphic remodeling of the larval nervous system and musculature of Phoronis pallida (Phoronida). AB - The structure of the larval nervous system and the musculature of Phoronis pallida were studied, as well as the remodeling of these systems at metamorphosis. The serotonergic portion of the apical ganglion is a U-shaped field of cell bodies that send projections into a central neuropil. The majority of the serotonergic cells are (at least) bipolar sensory cells, and a few are nonsensory cells. Catecholaminergic cell bodies border the apical ganglion. The second (hood) sense organ develops at competence and is composed of bipolar sensory cells that send projections into a secondary neuropil. Musculature of the competent larva includes circular and longitudinal muscle fibers of the body wall, as well as elevators and depressors of the tentacles and hood. The juvenile nervous system and musculature are developed prior to metamorphosis and are integrated with those of the larva. Components of the juvenile nervous system include a diffuse neural net of serotonergic cell bodies and fibers and longitudinal catecholaminergic fibers. The juvenile body wall musculature consists of longitudinal fibers that overlie circular muscle fibers, except in the cincture regions, where this pattern is reversed. Metamorphosis is initiated by the larval neuromuscular system but is completed by the juvenile neuromuscular system. During metamorphosis, the larval nervous system and the musculature undergo cell death, and the larval tentacles and gut are remodeled into the juvenile arrangement. Although the phoronid nervous system has often been described as deuterostome-like, these data show that several cytological aspects of the larval and juvenile neuromuscular systems also have protostome (lophotrochozoan) characteristics. PMID- 11868658 TI - Growth negatively impacts the life span of mammals. AB - A negative intraspecific relationship between growth and longevity was proposed in the early 20th century. Indeed, stunting the growth of rodents by restricting their food dramatically extended life span. Subsequently, however, the hypothesis that growth exacerbates aging rates fell into disfavor. Contributing to this was (a) the establishment of a positive relationship between body size and longevity interspecifically, (b) purported antiaging impacts of growth hormone, and (c) the fact that the longevity of even mature rodents that had completed growth was extended by dietary restriction. Furthermore, intraspecific analytical studies failed to provide any clear resolution. This article presents the first global analyses of maximal longevity versus maximum mature mass for laboratory rats and mice, based on a relatively comprehensive compilation of research across the 20th century. Peak body mass (which reflects juvenile growth rates) was negatively associated with longevity within both species. Proximal mechanisms for impacts of growth on longevity appear congruent with the free radical and immunological theories of aging. PMID- 11868657 TI - Comparison of the neuromuscular systems among actinotroch larvae: systematic and evolutionary implications. AB - A comparative analysis of the larval and presumptive juvenile neuromuscular systems among actinotroch larvae was performed using confocal laser microscopy with probes for F-actin and serotonin. Currently, there are two main categories of larval nervous systems based on the origin of the nerve fibers that innervate the larval tentacles. Characteristics of the serotonergic cells of the larval apical ganglion and juvenile nervous system have remained relatively conserved, but the structure of the secondary (hood) sense organ and the juvenile tentacles has diversified among species. Differences in larval musculature are mainly associated with differences in hood morphology. The presumptive, juvenile neuromuscular system is either integrated or separated from that of the larva based on the origin of the juvenile tentacles. Among species, the juvenile tentacles are made by remodeling the larval tentacles, developed from a basal tentacular thickening, or developed as a completely separate set in the larva. Differentiation of the neuromuscular structures of the juvenile tentacles is more diverse than their outward morphological characteristics would suggest. Importance of these larval characters is discussed in terms of current problems that exist within phoronid systematics. Evolutionary implications of these morphological characters are discussed among the phoronids, brachiopods, and related bilaterians. Overall, the integration or separation of larval and juvenile neuromuscular characters may yield insights into the evolution of lophotrochozoan body plans. PMID- 11868659 TI - Life history evolution and comparative developmental biology of echinoderms. AB - Evolutionary biologists studying life history variation have used echinoderms in experimental, laboratory, and field studies of life history evolution. This focus on echinoderms grew originally from the tradition of comparative embryology, in which echinoderms were central. The tools for obtaining and manipulating echinoderm gametes and larvae were taken directly from comparative embryological research. In addition, the comparative embryologists employed a diverse array of echinoderms, not a few model species, and this diversity has led to a broad understanding of the development, function, and evolution of echinoderm larvae. As a result, this branch of life history evolution has deep roots in comparative developmental biology of echinoderms. Here two main aspects of this relationship are reviewed. The first is a broad range of studies of fertilization biology, dispersal, population genetics, functional morphology, and asexual reproduction in which developmental biologists might take a keen interest because of the historical origins of this research in echinoderm comparative embryology. The second is a similarly broad variety of topics in life history research in which evolutionary biologists require techniques or data from developmental biology in order to make progress on understanding patterns of life history variation among echinoderm species and higher taxa. Both sets of topics provide opportunities for interaction and collaboration. PMID- 11868660 TI - AmphiFoxE4, an amphioxus winged helix/forkhead gene encoding a protein closely related to vertebrate thyroid transcription factor-2: expression during pharyngeal development. AB - The full-length sequence and developmental expression of amphioxus AmphiFoxE4 are described. Transcripts of the gene are first detected in the pharyngeal endoderm, where the club-shaped gland is forming and subsequently in the definitive gland itself. AmphiFoxE4 is closely related to vertebrate genes encoding the thyroid specific transcription factor-2 (TTF2), which plays an early developmental role in the morphogenesis of the thyroid gland and a later role in hormone-mediated control of thyroid function. In amphioxus, AmphiFoxE4 expression is not thyroid specific because the club-shaped gland, the only structure expressing the gene, is not homologous to the vertebrate thyroid; instead, the thyroid homologue of amphioxus is a specialized region of the pharyngeal endoderm called the endostyle. We propose that (a) the pharynx of an amphioxus-like ancestor of the vertebrates included a club-shaped gland that expressed FoxE4 as well as an endostyle that did not, and (b) the club-shaped gland soon disappeared in the vertebrate line of descent but (c) not before there was a homeogenetic transfer of FoxE4 expression from the club-shaped gland to the nearby endostyle. Such a transfer could have provided part of the genetic program enabling the endostyle to separate from the pharyngeal endoderm and migrate away as the rudiment of the thyroid gland. PMID- 11868661 TI - Heavy metals in organisms and sediments from Turkish Coast of the Black Sea, 1997 1998. AB - During the period 1997-1998, macroalgae, sea snail, mussel, fish and sediment samples were collected at different stations of the Turkish Black Sea coast in order to establish the concentration of selected heavy metals. Heavy metals analyzed were Cd, Co, Cr, Ni, Zn, Fe, Mn, Pb and Cu. The results showed that the Turkish Black Sea coast is facing heavy metal pollution. The metal concentrations in macroalgae, sea snail, mussel and sediment samples are very high. However, Cd, Pb and Cu concentrations in anchovy fish decreased, while Co, Fe, Zn, Cr, Mn and Ni contents had changed when compared previous data. The metal levels in macroalgae did not follow the same pattern as concentrations in sediments at the same station. On the other hand, concentrations of Cd, Co, Cr, Zn, Fe, Mn and Cu in sea snail, mussel and fish samples were related to sediment data in the examined stations. PMID- 11868662 TI - Regional radiation risk and vulnerability assessment by integration of mathematical modelling and GIS analysis. AB - The Kola Peninsula, Russian Arctic exceeds all other regions in the world in the number of nuclear reactors. The study was aimed at estimating possible radiation risks to the population in the Nordic countries in case of a severe accident in the Kola Peninsula. Two approaches were tested: (1) probabilistic analysis of modelled possible pathways of radionuclide transport and precipitation and (2) deterministic approach (case studies) for most possible or worst-case scenarios of modelled transport and deposition of radionuclides. For the general population, Finland is at most risk with respect to the Kola nuclear power plant (NPP) because of (a) relatively high population density or proximity to the radiation-risk sites and (b) rather high probability of an airflow trajectory there and precipitation. After considering the critical group, northern counties in Norway, Finland and Sweden appear to be most vulnerable. The case scenarios demonstrate that population in many counties in each country, both near and far away from a nuclear site, might be subject to high risk depending on the meteorological situation. PMID- 11868663 TI - Environmental geochemistry and pollution studies of Aliaga metal industry district. AB - The Aliaga metal industry district located 50 km northwest of Izmir City, in Turkey, includes many metal factories. The geology of the area is represented by Mesozoic flysch deposits and Cenozoic volcano sedimentary rocks. Tectonic elements are NE-SW and NW-SE trending faults and a W-NW trending fault direction that is important for water supply. Rock, soil, stream sediment, and water samples taken at various distances from the industrial area were analyzed for Fe, Ti, Mn, Cr, Pb, Cu, Ni, Zn, and Mo. According to the results, the elements in rocks are reasonable for the range of "Clarke" values, but in the soils and stream sediment, they comprise higher values than are acceptable for agricultural activities. Toxicity analyses were carried out in the drinking artesian water of Cakmakli village and wastewater samples of the factories and river water both of which are used for irrigation of the agricultural areas in and around the investigated area. The people should, however be made aware that this waste and river water is unacceptable for agriculture. Additionally, the contamination of seawater in Nemrut Bay is probably caused by contaminated river and underground water running to the sea. The chemical and toxicity analysis of drinking water samples show that they are above accepted standards and harmful. Previous air pollution studies (funnel gas emission analyses) also show that gas emission contains high amount of dust particles with high Fe, Zn, Pb, and Cu concentrations. As a result, all elements analyzed are of great importance relative to problems concerning contamination of the soil, stream sediment, ground surface water, and air by individual metals from uncontrolled processing of some metal factories. PMID- 11868664 TI - Microbial and nutrient pollution of coastal bathing waters in Mauritius. AB - The coastal pollution problem in Mauritius is exacerbated by the hydrogeology of the volcanic substratum. Bacterial contamination of bathing waters and nutrients, water temperature, salinity, and dissolved oxygen (DO) were monitored at three different spatial and temporal scales along the coastline of Mauritius during 1997-1998. Standard techniques for water sample collection and analysis set by the American Public Health Association [APHA. Standard methods for the examination of water and wastewater. 19th ed. Washington, DC: APHA, 1995.] were used at: (a) 16 sites around the island over a period of 7 months; (b) 12 stations along a recreational beach over an 18-month period; and (c) at an underground freshwater seepage point over 1 day. Total coliform (TC), faecal coliform (FC), and faecal streptococci (FS) contamination reported during all surveys varied randomly (e.g., with maximum densities in the ranges of 346-2020 TC, 130-2000 FC, and 180-1040 FS at one site) and at times exceeded the established EEC and Environment Protection Agency (EPA) standards for bathing water (e.g., in >90% of samples) to qualify for beach closure. Computed FC:FS ratios were used to pinpoint human faecal matter as the main source of contamination. Nitrate, phosphate, and silicate concentrations in seepage water were high (3600-9485, 38-105, and 9950-24,775 microg l(-1), respectively) and a cause for concern when compared with levels (5-845, 5-72, and 35-6570 microg l( 1), respectively) in cleaner lagoon water samples. Statistical analysis showed significant correlations (for TC and NO3: r=.75, P<.02; for TC and PO4: r=.779, P<.02; for TC and SiO4: r=.731, P<.05; for FC and NO3: r=.773, P<.02; for FC and SiO4: r=.727, P<.05; for FS and SiO4: r=.801 P<.01) between microbial densities and nutrients recorded, confirming the pathogen-contaminated water to be highly eutrophic. There is an urgency for Mauritius to properly address the issue of sewage treatment and wastewater discharge to safeguard its coastal environment, public health, and tourism expansion. PMID- 11868665 TI - Distribution analysis of airborne nicotine concentrations in hospitality facilities. AB - A number of publications report statistical summaries for environmental tobacco smoke (ETS) concentrations. Despite compelling evidence for the data not being normally distributed, these publications typically report the arithmetic mean and standard deviation of the data, thereby losing important information related to the distribution of values contained in the original data. We were interested in the frequency distributions of reported nicotine concentrations in hospitality environments and subjected available data to distribution analyses. The distribution of experimental indoor airborne nicotine concentration data taken from hospitality facilities worldwide was fit to lognormal, Weibull, exponential, Pearson (Type V), logistic, and loglogistic distribution models. Comparison of goodness of fit (GOF) parameters and indications from the literature verified the selection of a lognormal distribution as the overall best model. When individual data were not reported in the literature, statistical summaries of results were used to model sets of lognormally distributed data that are intended to mimic the original data distribution. Grouping the data into various categories led to 31 frequency distributions that were further interpreted. The median values in nonsmoking environments are about half of the median values in smoking sections. When different continents are compared, Asian, European, and North American median values in restaurants are about a factor of three below levels encountered in other hospitality facilities. On a comparison of nicotine concentrations in North American smoking sections and nonsmoking sections, median values are about one-third of the European levels. The results obtained may be used to address issues related to exposure to ETS in the hospitality sector. PMID- 11868666 TI - Emissions from heated indoor dust. AB - Settled indoor dust was collected from a university building, and the fine fraction was heated in the temperature range of 50-250 degrees C. Emissions of compounds were analysed and identified in a system consisting of a thermal desorption unit coupled to a gas chromatograph-mass spectrometer (GC-MS). The heating took place in both an oxidative (air) and an inert (He) atmosphere. The results indicate that oxidation of adsorbed compounds occurred, as well as decomposition of the dust itself. The emission in air started somewhere between 150 and 200 degrees C, with aldehydes and ketones as the main compounds. When heated in the inert atmosphere, semivolatile organic compounds (SVOCs) were emitted at temperatures above 150 degrees C. These compounds were not found in the air system, probably due to oxidation. Based on the obtained results we recommend that the temperature of hot surfaces in indoor environments should be kept below 150 degrees C to avoid oxidation of indoor dust and minimise the emissions. PMID- 11868667 TI - Hydroquinone: a general phagostimulating pheromone in termites. AB - The organization of termite societies depends predominantly on intraspecific chemical signals (pheromones) produced by exocrine glands, which induce and modulate individual behavioral responses. Here, the saliva-producing labial glands of termites were investigated with respect to their pheromonal role in communal food exploitation of termite colonies. From these glands, we identified for the first time hydroquinone (1,4-dihydroxybenzene) as a phagostimulating pheromone in the Australian termite species Mastotermes darwiniensis. Hydroquinone is released from the labial glands of termite workers and applied onto the food. It stimulates nestmates to feed at the spot of application and is, thus, employed to mark feeding sites. No synergistic effect with other identified labial gland compounds, such as glucose, inositol, and arbutin, was evident. Significantly, we show that termite species from all over the world, irrespective of taxonomic position and biological traits, produce and employ hydroquinone as phagostimulating signal. The use of the same chemical signal throughout an order is a unique phenomenon, not reported before in animals. Its possible biosynthetic pathway, ecological significance, and evolution are discussed. PMID- 11868668 TI - A novel function of the triterpene squalene in a tritrophic system. AB - Changes in apple leaf chemistry after infestation by leafminers and their effect on both host location and host habitat location of the generalist parasitoid Pholetesor bicolor were investigated. Chemical analysis of leaf solvent extracts from healthy and leafminer-damaged leaves revealed that herbivory increased the amount of the triterpene squalene (C30H50), whereas quantities of all other identified compounds were similar in both plant treatments. To assess the response of parasitoids to host location cues, contact bioassays were conducted with naive females. Results showed that parasitoids performed a characteristic ovipositional probing more often on the mine-damaged than on the healthy leaf. This behavior was triggered by a hexane extract of the mine-damaged leaf, but not by a healthy leaf extract. A synthetic mixture of the compounds identified in the extract triggered a similar response. A mixture devoid of squalene was not active, whereas squalene alone elicited the probing behavior. To assess the use of the identified compounds in habitat location, Y-tube olfactometer experiments were conducted with naive and experienced females. Results showed that squalene is not involved in habitat location and has no priming effect on P. bicolor. While other triterpenes are known to mediate habitat location of parasitoids, this is the first report in which a plant triterpene is shown to mediate host location of a parasitoid. The biological and ecological functions of squalene on all three trophic levels are discussed. PMID- 11868669 TI - Silphinene sesquiterpenes as model insect antifeedants. AB - Silphinene sesquiterpenes are established chrysomelid antifeedants. In this work, nine silphinene analogs, 11beta-acetoxy-5alpha-angeloyloxysilphinen-3-one (1), 11beta-acetoxy-5alpha-tigloyloxysilphinen-3-one (2), 11beta-acetoxy-5alpha isobutyryloxysilphinen-3-one (3), 11beta-hydroxy-5alpha-angeloyloxysilphinen-3 one (4), 11beta,5alpha-dihydroxysilphinen-3-one (5), 11beta,5alpha diacetoxysilphinen-3-one (6), 5alpha,11beta-diisobutyryloxysilphinen-3-one (7), silphinen-3,5,11-trione (8), and O-methyl-5-epicantabrenolic acid methyl ester (10), and a presilphiperfolane sesquiterpene (9) were tested against several divergent insect species, including the lepidopteran Spodoptera littoralis, the chrysomelid Leptinotarsa decemlineata, and five aphid species, and their antifeedant effects were compared with those of picrotoxinin, a GABA-antagonist, and thymol, an allosteric modulator for insect GABA receptors. All insects tested responded to at least one silphinene analog and/or GABA antagonist. Compound 3 and thymol were effective antifeedants against all species tested except S. littoralis, with varying potencies according to their feeding ecologies. The toxicity of these compounds was species-dependent and did not correlate with their antifeedant effect. PMID- 11868670 TI - Developmental, gustatory, and behavioral responses of leafroller larvae, Choristoneura rosaceana, to tannic acid and glucose. AB - Soluble sugars are essential nutrients generally perceived as phagostimulants to most insects studied. However, tannins are known as digestibility reducers, hence deleterious to caterpillar development, and as deterrents as well. Previous work demonstrated that larvae of the polyphagous oblique-banded leafroller, Choristoneura rosaceana, performed better when reared on a control + 0.5% tannic acid diet than on the standard control diet and that larvae reared on a control + 5% glucose diet had slower development and reduced survival. This study was designed to elucidate the behavioral and neurophysiological components of the larval responses to tannic acid and glucose. C. rosaceana larvae were reared individually from the first to the sixth instar on one of four different artificial diets: (1) control; (2) control + 5% glucose; (3) control + 0.5% tannic acid; (4) control + 5% glucose + 0.5% tannic acid. After 14 days, larvae reared on the control + 5% glucose diet had not developed past the fourth instar, whereas a considerable proportion of larvae reared on the control + 0.5% tannic acid diet had already attained the pupal stage. Insects reared on the control or the control + 5% glucose + 0.5% tannic acid diet had intermediate development. with most larvae in the fifth instar. In addition, once the mid-sixth instar was reached, the feeding preferences to 25 and 300 mM glucose, 25 mM tannic acid, and 25 mM glucose + 25 mM tannic acid over water were assessed in two-choice tests. Feeding affected preference. Control-reared insects preferred feeding on treatments containing glucose and were not deterred by tannic acid. However, larvae that had been exposed to tannic acid during their development were deterred by tannic acid and their glucose discrimination was impaired. The sensitivity to glucose was also examined from neurophysiological recordings by stimulating the sugar-sensitive cell (cell 1) on the lateral styloconic sensillum of the maxillary galea with increasing concentrations of glucose (1, 10, 25, 50, 100, 200, 300, and 500 mM). We also determined whether tannic acid was phagostimulatory, since insects develop relatively quickly on a diet containing this compound, by testing 1 mM tannic acid, 1 mM tannic acid + 300 mM glucose, and 300 mM glucose on the lateral styloconic sensilla. The traces indicated that 1 mM tannic acid was not detected by any of the four chemosensory cells in these sensilla. The combination of tannic acid and glucose produced no spikes from the sugar-sensitive cell, whereas a prominent spike activity resulted with 300 mM glucose. We concluded that, although C. rosaceana larvae develop faster on a tannic acid diet, this compound is not a phagostimulant. The converse is true for glucose; i.e., it stimulates the sugar-sensitive cell in the lateral styloconica in a concentration-dependent fashion. Previous dietary experience changes the sensory and behavioral responses of C. rosaceana to glucose. Our findings imply that not all compounds that are phagostimulatory are necessarily beneficial to an insect's fitness. Therefore, developmental studies should be interpreted in conjunction with behavioral and physiological data. PMID- 11868672 TI - In vivo volatile emissions from peanut plants induced by simultaneous fungal infection and insect damage. AB - Peanut plants, Arachis hypogaea, infected with white mold. Sclerotium rolfsii, emit a blend of organic compounds that differs both quantitatively and qualitatively from the blend emitted from plants damaged by beet armyworm (BAW; Spodoptera exigua) larvae or from uninfected, undamaged plants. Attackby BAW induced release of lipoxygenase products (hexenols, hexenals, and hexenyl esters), terpenoids, and indole. The plant-derived compound methyl salicylate and the fungal-derived compound 3-octanone were found only in headspace samples from white mold infected plants. White mold-infected plants exposed to BAW damage released all the volatiles emitted by healthy plants fed on by BAW in addition to those emitted in response to white mold infection alone. When BAW larvae were given a choice of feeding on leaves from healthy or white mold-infected plants, they consumed larger quantities of the leaves from infected plants. Exposure to commercially available (Z)-3 hexenyl acetate, linalool, and methyl salicylate, compounds emitted by white mold-infected plants, significantly reduced the growth of the white mold in solid-media cultures. Thus, emission of these compounds by infected plants may constitute a direct defense against this pathogen. PMID- 11868671 TI - Evidence of an assembly pheromone in the black-legged deer tick, Ixodes scapularis. AB - The responses of Irodes scapularis (Acari: Ixodidae) nymphs and adults to extracts of cast larval skins were tested in a Petri dish bioassay. Assembly was elicited in nymphs and adults in the presence of skins, exudate from ticks, and filter paper exposed to ticks compared to untreated controls. Assembly was noted by 1 hr after exposure with little change between 1 and 24 hr. The assembly response increased in the presence of an increased number of skins. Similar assembly was elicited in nymphs and adults in the presence of cast larval skins and a saline (0.95% NaCl) skin extract. Methanol and hexane extracts were not attractive. When chemical standards were tested against nymphs, they responded to guanine, uric acid, hypoxanthine, xanthine, inosine, and hematin. Adults were tested against guanine, inosine, and xanthine, and all elicited significant assembly. Responses of nymphs increased significantly with increase in dose of uric acid and guanine. Responses of nymphs to a mixture of guanine, xanthine, and adenine (25:1:1 ratio) were similar to responses to cast skins. This study provides the first evidence of an assembly pheromone in I. scapularis. PMID- 11868674 TI - Oviposition responses by hessian fly, Mayetiola destructor, to wheats varying in surfaces waxes. AB - Leaf waxes are known to contain oviposition stimulants for Hessian fly. In dual choice tests comparing seedlings of each of three nonglaucous lines of wheat with its corresponding glaucous line, Hessian flies laid similar numbers of eggs on each genotype. However, when plants from these genotypes were tested at the flag leaf stage. Hessian flies deposited 25-100% more eggs on the nonglaucous genotype compared to normal wax genotype in each pair. Leaf waxes extracted from the genotypes in one of these pairs (Avalon and nonglaucous Avalon) at the seedling and the flag-leaf stage and tested on paper leaf models elicited oviposition responses similar to those observed on the intact plants. Scanning electron microscopy showed differences in crystal density on nonglaucous and glaucous genotypes only at the flag-leaf stage. Gas chromatography-mass spectrometry of the leaf wax extracts showed that fly oviposition responses were associated with differences in the chemical composition of the waxes. This is the first report of Hessian fly oviposition responses varying among genotypes of wheat with different surface wax composition. PMID- 11868673 TI - Evaluation of synergism in the feeding deterrence of some furanocoumarins on Spodoptera littoralis. AB - The phagodepression activity of five furanocoumarins (FC), bergapten, xanthotoxin, psoralen, imperatorin, and angelicin, has been studied against larvae of Egyptian cotton leafworm Spodoptera littoralis (Boisduval) (Lepidoptera: Noctuidae) using a leaf-disk choice bioassay. The dose range used was 0-10 microg/cm2 for linear furocoumarins and 0-30 microg/cm2 for angelicin, the angular furanocoumarin. Dose-feeding deterrency activity correlations were governed by various sigmoidal functions, except in the case of imperatorin and bergapten, which had dose-response curves showing irregular traces with two maxima. All five FCs induced various degrees of phagodepression in S. littoralis; at 3 microg/cm2, the relative feeding deterrence was bergapten = xanthotoxin > psoralen = imperatorin = angelicin. Structure-activity correlations indicated that a methoxy group on C-5 or C-8 enhanced the activity. Comparison of experimental feeding deterrence of binary mixtures of imperatorin with xanthotoxin, bergapten, or psoralen at various relative concentrations with the calculated additive activity of each combination indicated that a proportion of 40% or more of imperatorin may exert a greater antifeedant effect on S. littoralis than the sum of individual compounds. The effect of angelicin also was examined in combination with psoralen, both of which are present in Psoralea plants. Their mixtures yielded a clear synergistic effect in the 20-80% angelicin range (coactivity coefficient = 124-133). By contrast, the effect of angelicin on xanthotoxin was only additive. In the dose-response curves of imperatorin/bergapten combinations, synergism was found at >60% imperatorin relative concentration and 1 microg/cm2, whereas lower proportions led to antagonism. The threshold between the two opposing effects was found to depend on the total FC concentration employed. Some natural systems contain FCs in the range of synergistic proportions recorded here and, thus, may have been produced by the host to increase its defensive effect at a lower metabolic cost. PMID- 11868675 TI - Comparing the variation of needle and wood terpenoids in Scots pine provenances. AB - We determined variation in both the concentration and composition of terpenoids in needles and wood within nine Scots pine (Pinus sylvestris) provenances. Seedlings of different provenances representing a 1200-km N-S transect from Estonia to northern Finland were cultivated in Suonenjoki nursery field, central Finland, for seven years. Growth of seedlings and the number of vertical resin ducts in wood were also determined. alpha-Pinene and 3-carene were the major monoterpenes both in the needles and wood. The total monoterpene concentration was about five times higher in the needles than in the wood. A strong positive correlation was found between proportional quantities of several terpenes of the needles and wood, particularly for 3-carene, sabinene, and terpinolene. The needles contained both labdane-type and tricyclic resin acids, whereas the wood contained only tricyclic ones. The wood had a four times higher tricyclic resin acid concentration than the needles. The highest total monoterpene concentration in the needles and in the wood occurred in the most northern Muonio provenance and in the most southern Saaremaa provenance plants, respectively. The amount of high 3-carene genotype trees decreased among the northern provenances. The wood of the most northern Muonio provenance showed the lowest total resin acid concentration, but provenance did not affect total tricyclic resin acids in the needles. Korpilahti provenance trees from central Finland had the best growth in height. In addition, Korpilahti and Ruokolahti provenance trees showed largest radial growth of stem and smallest number of vertical resin ducts. The results suggest that especially the proportional quantity of 3-carene in the needles could be used in estimating the amount of this compound in the wood and vice versa. PMID- 11868676 TI - Why phenolic acids are unlikely primary allelochemicals in rice. AB - Allelopathy in rice (Oryza sativa, L.) effective against weeds has been found in about 3.5% of tested rice germplasm in both laboratory and field experimentation. However, the allelochemicals responsible for growth inhibition of rice-associated weeds have not yet been identified. In the literature, phenolic acids are often mentioned as putative allelochemicals. If phenolic acids commonly reach growth inhibitory concentrations in rice ecosystems, it must be expected that the degree of tolerance to phenolic acids will vary among traditional rice cultivars or plant species adapted to rice environments having inherently different phenolic acid concentrations. Phenolic acids concentrations are normally greater in submerged than in aerobic soils. A dose-response study, however, showed that seedlings of rice cultivars adapted to submerged anaerobic soils did not have higher level of tolerance against p-hydroxybenzoic acid than did seedlings of varieties adapted to aerobic upland soils. Moreover, traditional rice cultivars had no greater tolerance than did improved cultivars that were recently bred for traits other than tolerance of phenolic acids. Similarly, there were no differences in tolerance of p-hydroxybenzoic acid between two Echinochloa weed species adapted to either anaerobic or aerobic growth conditions. Thus, neither the rice cultivars nor weed species had evolved different tolerance levels against the phenolic acid. However, all rice cultivars had significantly greater tolerance of p-hydroxybenzoic acid than did either weed species. In a second experiment, the rates at which rice plants released phenolic acids into solution cultures were measured for at least one month, the time period of greatest allelopathic activity following planting under field conditions. The maximum release rate of phenolic acids during the first month of growth was approximately 10 microg/plant/day. At a conventional plant density, the release rate of phenolic acids would be approximately 1 mg/m2/day. This order of release rate cannot provide concentrations remotely close to phytotoxic levels determined for these rice cultivars and weed species. The results presented in this paper do not preclude the possibility that phenolic acids might be one component in a mixture of chemicals that, when present simultaneously, are allelopathic. PMID- 11868677 TI - Mature atlantic salmon (Salmo salar L.) male parr are attracted to ovulated female urine but not to ovarian fluid. AB - The behavioral responses to urine and ovarian fluids from conspecific and heterospecific ovulated females were studied in mature Atlantic salmon (Salmo salar) male parr in a two-choice fluviarium. The males reacted differently to the stimulants. They spent more time in water scented by urine from salmon or brown trout (Salmo trutta L.) females compared to the time in water with ovarian fluids from salmon females. Furthermore, the males were attracted to salmon female urine (different from an indifferent reaction). Males exposed to urine of either species had higher plasma concentrations of testosterone (T) compared to unexposed controls. Measurement of the concentrations of prostaglandin F2alpha (PGF2alpha) and its major metabolite 15-ketodihydroprostaglandin F2alpha (15 ketodihydro-PGF2alpha) showed that the concentrations of the substances were higher in ovarian fluids of both species compared to those in urine. PGF2alpha showed a greater difference between ovarian fluid and urine than its major metabolite. The results suggest that urine of both species, in contrast to ovarian fluid, contain substances that attract mature Atlantic salmon male parr and that the active substances are neither PGF2alpha nor 15-keto-PGF2alpha. PMID- 11868678 TI - Gas-chromatographic analyses of the subcaudal gland secretion of the European badger (Meles meles) part I: chemical differences related to individual parameters. AB - In many species, chemical signals are important in the context of intraspecific communication. European badgers (Meles meles) use the secretion of their subcaudal gland to mark both the environment and conspecifics. In this paper, we investigated the chemical composition of subcaudal gland secretions and how it varies among individuals according to group membership, season, sex, age, body condition, and reproductive status. We analyzed 66 samples by gas chromatography using a mass-spectrometer as detector. We found 110 different components, of which 21 were present in every profile. The chemical composition of the secretions proved to be highly individual-specific, but no single peak was found to be typical for one of the investigated categories (season, sex, etc.). Analyses of the relative area (% area) that every peak contributed to the overall profile area showed that, in general, group members have more similar profiles than do badgers from different groups. Composition varied over seasons and between sexes and was influenced by age, body condition, and reproductive status. We, therefore, conclude that the secretion not only encodes group membership and individuality as previously assumed, but might also convey information about fitness-related parameters. PMID- 11868679 TI - Gas-chromatographic analyses of the subcaudal gland secretion of the European badger (Meles meles) part II: time-related variation in the individual-specific composition. AB - Individuality in body odors has been described in a variety of species, but studies on time-related variation in individual scent are scarce. Here, we use GC MS to investigate how chemical composition of subcaudal gland secretions of European badgers (Meles meles) varies over days, seasons, and from year to year, and how secretions change with the length of time for which they are exposed to the environment. Samples were divided into subsamples--one was frozen immediately and the remaining ones frozen after 2, 6, 12, 24, and 48 hr, respectively--and many individual-specific characteristics of the scent-profiles remained stable over time. However, two components were negatively correlated with time, thus providing the possibility to determine the age of scent marks. The low variation found in scent profiles of samples collected from the same individual three days apart showed that the individual-specific scent is a true characteristic of the respective badger, and that trapping and subsequent sampling have little effect on the composition of subcaudal gland secretions. Long-term variation (i.e., over one year) in individual subcaudal scent profiles is not continuous, but periods of relative stability are followed by periods of rapid change, that can be related to badger biology. Annual variation between samples collected from the same individuals in winter 1998 and winter 1999. and in spring 1998 and spring 1999 was lower than seasonal variation. Therefore, the results of this study indicate the potential of an individual-specific scent signature in the subcaudal gland secretions of badgers evidencing that individual recognition is of high importance in this species. PMID- 11868680 TI - Dufour gland contents of ants of the Cataglyphis bicolor group. AB - The species of desert-dwelling ants of the Cataglyphis bicolor (Hymenoptera: Formicidae) group are difficult to distinguish by morphological features. Analysis of the secretion from the Dufour glands of workers of a number of colonies was undertaken to see if it provided a clear test of species. Linked 6c ms showed in all samples straight and branched-chain alkanes, linear alkenes, ketones, aldehydes, acetates, and a group of C22 to C28 esters not previously identified in this genus. Contents of the Dufour glands of C. savignyi from Tunisia and Egypt were similar, and comprised straight and branched-chain alkanes, alkenes and small amounts of esters. C. bicolor from Tunisia contained compounds similar to C. savignyi but was distinguished from the latter by larger amounts of the esters. The major compound in the glands of C. viaticus was tridecane, in contrast to the pentadecane of other species. It also contained a branched alkane, 3-methyltridecane as a major component. Branched-chain esters and a wide variety of acetates were also found in this species. C. diehlii had a limited range of compounds, with branched alkanes almost completely absent and high proportions of pentadecene and dodecyl acetate. C bombycinus, a sympatric species. but recognized as not belonging to the bicolor group by its different mandibular gland substances, was notable in having butanoate esters in its Dufour glands. Despite these differences among species, both the great variability of individuals from a single colony and the among between conspecific colonies make species diagnosis from a few individuals difficult, in contrast with postpharyngeal glands, which, as recently reported, give a clearer indication of species. PMID- 11868682 TI - Patient:nurse ratios in critical care--time for some radical thinking. PMID- 11868681 TI - Mammalian exocrine secretions. XVII: chemical characterization of preorbital secretion of male suni, Neotragus moschatus. AB - Gas chromatographic and gas chromatographic-mass spectrometric techniques were employed to identify 83 compounds, including alkanes, alkenes, aldehydes, 2 methylalkanes, carboxylic acids, 1-alkyl formates and alken-1-yl formates, benzoic acid, and cholesterol, in the preorbital secretion of the male suni, Neotragus moschatus. Dimethyl disulfide derivatization and lithium aluminum hydride reduction were used to determine the position of double bonds and to confirm the identity of the functional groups in some of the constituents of the secretion. PMID- 11868683 TI - The influence of focus group-oriented supervision on intensive care nurses' reflections on family members' needs. AB - The aim of this study was to explore intensive care nurses' experiences of focus group-oriented supervision with particular reference to family members' needs. In addition, the aim was to focus on the intensive care nurses' perceived change in their insight into caring for patients and family members in an intensive care unit. Four themes were constructed: increased perception of and response to the family members' needs; increased self-insight related to the therapeutic use of oneself in the relationship with patients and their family members; nurses' reflection on factors that increased their competence; and increased creativity. In conclusion, focus group-oriented supervision increased the intensive care nurses' understanding of their role. This has consequences not only for the family members but also for the teamwork in the intensive care unit, where this type of work is common. PMID- 11868684 TI - Transfer anxiety: preparing to leave intensive care. AB - There is much literature to substantiate the inadvertent emotional and psychological trauma associated with critical care areas. However, alongside this, there is a growing body of knowledge to show that these intense and specialized areas are actually perceived as secure, safe and familiar environments by some patients and family members. Transfer from the intensive care unit is not always perceived in a positive light and often the transition is dreaded by both the patient and his family. The evidence would suggest that discharge from specialized care environments can actually be as traumatic as admission. This phenomenon has become known as transfer anxiety, relocation anxiety, or translocation anxiety. There is the possibility that transfer may induce stress or distress in some patients, especially when routines, environments and/or invasive monitoring procedures are altered or ceased without prior knowledge, preparation or adequate explanation. If healthcare personnel fail to identify and meet the psychological needs of patients and families relocating from these areas, the detrimental effects may extend far beyond discharge from ICU. For relocating patients, transfer from the ICU can be presented as a positive step. However, treatment to minimize transfer anxiety will only be successful when all healthcare personnel recognize and react positively to the psychological factors that affect patients adversely. PMID- 11868685 TI - Maintaining patient dignity in intensive care settings. AB - Maintenance of the dignity of intensive care unit (ICU) patients, particularly minimizing exposure of genitalia, may be problematic. The aim of the study was to develop strategies to maximize the dignity of ICU patients using an action research methodology. The first stage assessed current practice through 62 hours of non-participant observation of patient care. Patient dignity was maintained in almost one-third of observed cases. However, more intimate areas such as bosom and genitalia were exposed in over 40% of the incidents. Whilst screens were fully used in over one-third of exposure incidents, full screening did not occur for all or part of the remaining incidents. Significant factors (P < 0.05) influencing exposure included gender and age. Female and younger (< 60 years) patients were more likely to be exposed; older patients (> 70 years) were less likely to be screened when exposed. The next stage involved identification of solutions to the problem of inappropriate patient exposure through the medium of a multi-disciplinary focus group. The focus group recommended raising staff awareness and documentation of situations that may compromise maintenance of dignity. The final stage of the study involved an audit of these recommendations. The main audit findings were more adequate clothing of patients plus a high level of staff awareness of patients' dignity needs. PMID- 11868686 TI - Update on the pathology of AIDS. AB - The acquired immune deficiency syndrome (AIDS) is the result of a human immunodeficiency virus (HIV) infection damaging the cell-mediated immune system. HIV infection is a zoonosis that has affected man for at least 50 years, though the disease has become pandemic only in the last 2 decades. A wide range of opportunistic infections (Ols) and tumours develop; additionally, HIV directly damages some organs. The patterns of opportunistic diseases (ODs) are significantly different in different parts of the world, depending on the local prevalences of latent and acquired infections and on the survival of HIV-infected patients. OD patterns change as peoples migrate. Recent highly active anti retroviral (HIV) chemotherapy prevents many of the common Ols, but also induces a new range of toxic pathological damage, e.g. the liver, and lipid metabolism. Longer survival permits development of new HIV-related diseases, e.g. the lymphoproliferative disorders. The pathology of HIV/AIDS is not static but changing. PMID- 11868687 TI - Nurse-led weaning from mechanical ventilation: where's the evidence? AB - BACKGROUND: Several authors have suggested clinical protocols as a means of shortening ventilation time and the important role of the nurse in reducing ventilation time has also been highlighted. Despite the many references in the literature to reducing weaning times using clinical protocols, it is not clear whether nurse-led weaning strategies hasten weaning from mechanical ventilation compared with physician-led care. OBJECTIVE: to systematically review the published literature to answer the question: Does nurse-led weaning from mechanical ventilation reduce duration of ventilation compared with doctor-led care? DATA SOURCES: MEDLINE, CINHAL, EMBASE, Cochrane Library, Best Evidence, hand search, expert opinion, controlled trials register. STUDY SELECTION: randomized controlled trials (RCTs) and cohort studies where nurses or respiratory therapists lead the weaning from mechanical ventilation; duration of ventilation must be stated. DATA EXTRACTION: performed by the author, who extracted data on statistical significance of the difference in duration of ventilation between control and trial groups, complication rates (such as reintubation) and mortality. DATA SYNTHESIS: statistical synthesis was not attempted but narrative synthesis was performed. RESULTS: only 1 randomized controlled trial and 2 cohort studies were found where nurses or respiratory therapists led weaning using a protocol. Two studies showed reduction in ventilation time without additional complications; the third study had the weakest evidence, because it was retrospective, and showed no difference between control and treatment groups. CONCLUSIONS: There is limited evidence suggesting that nurse-led weaning may reduce ventilation time; however, it is not clear whether it was the nurse-led aspect or the clinical protocol that produced the effect. PMID- 11868688 TI - Activated protein C in severe sepsis. AB - A recent large trial has shown that recombinant activated protein C reduced mortality in severe sepsis. This is the first real advance in the pharmacotherapy of sepsis since the introduction of antibiotics in the last century. PMID- 11868689 TI - POCT regulatory compliance: what is it and how does it impact you? AB - Point-of-care testing (POCT), ancillary testing, bedside testing, and near patient testing all refer to the same thing--a form of testing that enables caregivers to determine results of lab tests at the bedside or near a patient on the floor. Performance of this type of testing has increased dramatically in the past several years. Why? There are several reasons. First, many clinicians want results Stat. They want to know immediately where the patient stands as far as blood glucose results, electrolytes, pregnancy tests, hematocrits, and ACTs, to name just a few. Physicians and nurses want to be able to make decisions quickly based on patient results and do not want to have to wait for the answers. Second, many vendors have been releasing better, simpler, more efficient devices that meet the needs of the caregiver by providing lab results in a timely manner. Third, regulatory bodies are recognizing and embracing this type of testing and are creating more complex checklists for compliance when performing these tests. This article focuses on the regulatory component of POCT to educate and familiarize individuals with an overview of the regulatory/accrediting bodies enforcing compliance in the hospital setting. PMID- 11868690 TI - Benefits of a joint nursing and laboratory point-of-care program: nursing and laboratory working together. AB - Critical care nurses contend daily with high pace, crisis-care, and dealing with patients, physicians, coworkers, and patients' family members. A point-of-care testing (POCT) program can assist with the quick decision making required of today's critical care nursing staff. By meshing the backgrounds of nursing and the central laboratory into one workable philosophy, POCT can ensure optimal patient care. PMID- 11868691 TI - The who, what, why, and how of point-of-care testing: one city's experience in the Group for South Texas Ancillary Testing (GSTAT). AB - San Antonio, Texas, is the eighth largest city in the United States and has several distinct hospital systems. For-profit and not-for profit hospitals exist in the city along with a county hospital and multiple military institutions. This article compares and contrasts the structure of point-of-care testing (POCT) programs in the five largest systems. Specifically, inpatient glucose testing is examined. This comparison includes the nursing responsibilities and fiscal components of the programs, including billing. PMID- 11868692 TI - Development of a universal connectivity and data management system. AB - Point-of-care testing (POCT) is an increasingly popular method of delivering laboratory testing. Management of POCT is challenging given the variety of devices, locations, and staff that need to be coordinated to ensure quality results and meet regulatory guidelines. Electronic capture and transfer of data are preferred for managing POCT, but there is currently no standard method of connecting different devices. Johns Hopkins Medical Institutions (JHMI) developed a common data management system with interfaces to all of its POCT devices. All POCT data are collected in one database and analyzed in a similar fashion. Where data were once collected by carrying laptops to each nursing unit, the POCT devices can now connect directly to the database over the Internet. Algorithms have been created to automate the data analysis and review process. Over the several years that this software has been used, JHMI has experienced improved quality, accuracy, and management of its POCT program. The labor saved by increased automation of data review is refocused on enhancing the performance and scope of the program. Current connectivity and analysis algorithms have future application to remote consultation, management of home self-monitoring patients, and examination of real-time data. PMID- 11868693 TI - Point-of-care testing at the millennium. AB - Point-of-care testing (POCT) is a major force in the future evolution of hospital care, with prospects for even greater expansion of accessibility, speed, and also, hopefully, accuracy of results. New developments in POCT technology will predictably occur in three areas: connectivity, test menu expansion, and noninvasiveness. Connectivity for POCT devices has evolved from point-of-service workstations to standardized POCT data transmission protocols to remote roaming wireless connectivity with automatic data capture. POCT test menus will continue to expand, with more coagulation testing, chemistries, and infectious screening, but also on-site drug screening, intraoperative hormone levels, and microchip DNA diagnostics. Noninvasive POCT will expand beyond the GlucoWatch glucose monitor and the Bilichek noninvasive bilirubin monitor to noninvasive CBCs and Pap smears. PMID- 11868695 TI - The value of point-of-care testing when instituting an insulin drip protocol. AB - The critically ill patient's response to stress is to increase production of glucose; his can lead to hyperglycemia as insulin releasing factors become overloaded. Although parenteral insulin can aid in preventing complications of hyperglycemia by maintaining a euglycemic state, care must be taken to prevent hypoglycemia. The safest way to prevent large fluctuations in blood glucose levels is frequent monitoring of blood glucose levels via bedside glucose testing. As blood glucose levels fluctuate, the critical care nurse may titrate an insulin drip without specific orders for how much to increase or decrease the amount of insulin infusing. By developing a protocol, including a formula to calculate the amount of insulin to infuse, based on blood glucose levels, the critical care nurse can safely, competently, and accurately maintain blood glucose levels under 250 mg/dL. PMID- 11868694 TI - Point-of-care testing in the United Kingdom. AB - Point-of-care testing in the United Kingdom is currently at the embryo stage of development, and is approximately 10 years behind the United States in incorporation and application into clinical practice. Safeguards to ensure proper use are not in place within the UK at present, and there is no legal requirement for POCT users to be proficiency tested by an internal or external body before they can routinely use the equipment. Internal quality control and external quality assessment are not statutory requirements within the UK, leading to unease regarding the extent and range of point-of-care tests offered and whether the results are a true biochemical representation of the patient's clinical condition. Cost-benefit analysis is very sparse, but faster turnaround times can only lead to more rapid treatment of the patient, which will lead to fewer clinical complications. This will result in decreased hospital admissions by the emergency department and reduced length of hospital stay in ward-based patients. National Health Service (NHS) accountants will see this, as the way forward, as fewer patients being admitted into hospital and length of stay reduction will result in decreased expenditure by the service provider. PMID- 11868696 TI - Cardiac markers and point-of-care testing: a perfect fit. AB - Biochemical markers are of increasing importance in diagnostic strategies for ruling in and ruling out acute myocardial infarction (AMI), particularly when electrocardiographic (ECG) findings do not allow a diagnosis. Point-of-care testing (POCT) or "near-patient" testing allows for diagnostic assays to be performed at the site of patient care delivery. The biochemical markers that are commonly used by physicians to aid in the diagnosis of AMI are myoglobin, CK-MB, troponin I, and troponin T. Currently available POCT assays possess comparable diagnostic performance to laboratory-based cardiac marker assays. This provides an opportunity for POCT to evolve into the standard of care for evaluating the greater than 6 million American patients presenting with chest discomfort. Full acceptance of this relatively new technology will not be realized until users reach a comfort level where utilization of these devices is foolproof and they have faith in the results. PMID- 11868697 TI - New strategies in the prevention of restenosis. AB - Restenosis is a common and serious complication following angioplasty and stent implantation in patients with arterial vascular disease. Restenosis is a form of intimal hyperplasia. Endothelin-1 (ET-1) and vascular endothelial growth factor (VEGF) stimulate intimal hyperplasia and may play a role in restenosis. ET-1 and VEGF may act in concert in promoting restenosis following mechanical injury to the vessel wall in angioplasty and stent implantation. An understanding of their mechanism of action may lead to more effective methods for preventing restenosis. ET-1 receptor antagonists may play a prominent role in prophylaxis. PMID- 11868698 TI - Liver transplantation and nursing care in Israel. AB - The Liver Unit in Hadassah Medical Center has been following patients before and after transplantation in a process combining medical and nurse care. Since 1991, 145 patients have been studied. This article reviews the study methods associated with the liver transplantation process, including etiology of basic liver disease, drug adverse effects, complications, and survival rates. The value of the nursing coordinator in liver transplantation is discussed. PMID- 11868699 TI - Implementing an effective point-of-care testing program. AB - The implementation and management of a point-of-care testing program requires the effective cooperation of nurses, pathologists, other physicians, technologists, and vendors. These individuals working collectively must establish the clinical and financial bases for establishing a program, for managing its daily operation, and for assuring its ongoing quality and value to patient care. PMID- 11868700 TI - Measurement issues in promoting continued competence. AB - PURPOSE: Promoting continued competence in nursing practice has always been of concern to staff development and continuing education educators. Recently it has reached new levels of significance. As a result of the proposals made by the Pew Commission Taskforce on Health Care Workforce Regulation and the National Council of State Boards of Nursing, the profession is re-examining the question, "How do we promote and assure continued competence?" The purpose of this article is to examine the measurement issues related to answering this question. FINDINGS: The nursing literature provides several definitions of competence, but none are operational. To measure a concept it must be operationalized. Mandatory continuing education, peer review, and practice or process audits have been used to assess continued competence. The measurement issues involved in assessing competence include selecting an appropriate measurement paradigm, selecting accurate measurement instruments, and interpreting the measurement data. IMPLICATIONS: Staff development educators and nurse managers should collaborate in the establishment of standards of practice, the identification of essential competencies specific to the setting, and the development of appropriate methods of measurement of these competencies. PMID- 11868701 TI - Development of continuing nursing education offerings for the World Wide Web. AB - Nurses are seeking continuing professional development that is easily accessible, convenient, and available at any time and any place. As nurses have increasing access to Internets and Intranets at home and their workplace, courses for continuing nursing education must be available to meet this need. This article discusses the planning, implementation, and evaluation of continuing nursing education (CNE) on the World Wide Web. The article explains how to develop a strategic plan, develop course offerings, select technology tools to support teaching and learning, and market and evaluate the courses. PMID- 11868702 TI - There really is a difference: home care competencies. AB - BACKGROUND: More nurses are turning to home care agencies as sources of employment because of mergers, alliances, and increased emphasis on outpatient services. The unique differences in the home care nursing role necessitate adequate orientation and ongoing competency validation. METHOD: A review of relevant nursing literature was conducted. The author also relied on her experiences as a home care educator. RESULTS: Nurses who enter the specialty of home care nursing can expect to learn specific competencies that are unique to home care. CONCLUSION: Specific competencies unique to home care are proposed. There is scant published research regarding home care competencies to date. PMID- 11868703 TI - The state board of nursing and its role in continued competency. AB - Many agencies and individuals are involved with nurse competency, but within this cooperative effort, state boards of nursing have the unique role of assuring the public, through the issuance of licenses, that nurses are competent. The purpose of the board of nursing, as defined by public lawmakers, is to protect the public. There are public members on most boards. Boards of nursing are involved with nurse competency at the time of the nurse's initial entry into practice, on an ongoing basis at the time of license renewal, when the nurse reenters practice after a time of voluntary absence, and after disciplinary action of the nurse. The participation in continuing education activities is one major method for nurses to meet continued competency requirements. PMID- 11868704 TI - Texas nurses respond to mandatory continuing education. AB - This study investigated Texas nurses' attitudes toward mandatory continuing education, and their perceptions of skill improvement, knowledge enrichment, and improvement of health care to the public as a result of participation in continuing education programs. Seventy-two percent of the respondents had a positive attitude toward mandatory continuing education. Significant relationships were present between the attitudes and the perceptions of psychomotor skills improvement, cognitive improvement, affective skills improvement, and health care to the public. Four variables were predictive of the attitude toward mandatory education. In addition, the nurses preferred lecture or discussion formats and independent providers of continuing education. PMID- 11868705 TI - Mandatory continuing education: past, present, and future trends and issues. AB - Mandatory continuing education (CE) for nurses continues to be a viable means by which nurses can remain competent in the face of ever-increasing knowledge and technology in the health care setting. A multiplicity of CE providers, offering a variety of teaching approaches, exists to ensure nurses remain competent and advance in their fields of practice. Presently, more research must be conducted to measure the effectiveness of CE programs on patient care outcomes. Scholarly investigation may assist in justifying mandatory CE for nurses if positive patient care outcomes can be identified. Several states are regulating professional nursing practitioners by requiring CE. Although some states have not endorsed mandatory CE for nurses, it is likely that more state boards of nursing will continue the slow but steady trend of requiring CE for relicensure- especially if research establishes positive patient care outcomes. PMID- 11868706 TI - Simplified recordkeeping: recordkeeping in an approved provider status unit. AB - Management of a continuing nursing education (CNE) Approved Provider Status Unit is an important, yet time-consuming process for the CNE Program Administrator. With proper instruction and clear delineation of recordkeeping role responsibilities, the CNE Program Administrator can responsibly delegate specific time-consuming functions to designated clerical personnel. PMID- 11868707 TI - Graduate nurses' experiences of interactions with professional nursing staff during transition to the professional role. AB - As new members of the nursing profession, graduate nurses (GNs), in their transition to the professional role, have a variety of experiences with registered nurses. These experiences give rise to diverse thoughts and emotional responses that may have a significant impact on nurses' own professional development and socialization to the professional role. Literature documenting GNs' perceptions and feelings regarding their orientation to the professional role is scarce. This study involved 16 nurses' descriptions of their own experiences, thoughts, and feelings that arose from interactions with nurse colleagues during their first year of professional nursing practice. There was a lack of consistency regarding the way in which GNs were assisted in this important transitional teaching and learning experience. Understanding GNs' perceptions of the process of transition to the professional role holds tremendous value for nursing education. This understanding could serve as a guide in the development of positive and appropriate orientation strategies as we strive to imbue nursing graduates with the professional values, attitudes, and behaviors that are essential in achieving outcomes reflective of quality patient care. PMID- 11868708 TI - Group involvement in decision-making: a pilot study. AB - BACKGROUND: The Vroom-Yetton-Jago Leadership Model served as a model to determine if leaders could be taught the appropriate level of group involvement in decision making. METHOD: A convenience sample of 27 health care leaders who attended a 90 minute class offered in one institution were eligible for inclusion in the study and pre- and post-performance on similar case studies was measured. RESULTS: Using a paired t test, results for this sample were statistically significant (t [21] = 6.02, p < .001). CONCLUSION: These results suggest that participating in a class on the use of this leadership model may help leaders gain the skill needed to appropriately delegate decision-making to groups. PMID- 11868709 TI - Development of human resources in nursing: a collaborative initiative in CPR. AB - BACKGROUND: An international collaborative project between Jordanian and Canadian University nursing faculty was developed to enhance staff development for practicing nurses in Jordan. A needs assessment indicated a lack of knowledge and performance among staff nurses in emergency situations. METHOD: Using the "train the-trainer" concept, cardiopulmonary resuscitation (CPR) workshops were organized and implemented in three government and one military hospital. University faculty developed and conducted the first workshops when local trainers were prepared to provide the program in their institutions. RESULTS: Local trainers currently provide CPR workshops on an ongoing basis in their institutions. To date, 10 hospitals and one health center have requested CPR workshops for nursing staff. CONCLUSION: The CPR workshops met the needs of the hospital nursing community and strengthened the partnership between the university faculty and local community staff nurses. PMID- 11868710 TI - Continuing education for nurses in Tianjin Municipality, the People's Republic of China. AB - BACKGROUND: A descriptive survey examined continuing education experiences of hospital nurses working in Tianjin Municipality, the third largest municipality in The People's Republic of China. METHOD: Fourteen hospitals and two hundred nurses were selected randomly. RESULTS: Over two thirds of the nurses had attended continuing education events in the previous few years. Learning experiences included on-site and off-site workshops; associate degree courses; and teaching strategies of mostly lectures, films and videos. Major barriers discouraging nurses from participating included lack of time, cost, distance, and being denied permission to attend. Nurses working in rural and suburban hospitals reported less access to continuing education opportunities than nurses in urban hospitals. Ninety-six percent of respondents reported they had made changes in their clinical practice as a result of the continuing education activities. CONCLUSION: Strategies to reduce barriers to continuing education and future research examining the impact of continuing nursing education on clinical practice in China need to be developed. PMID- 11868711 TI - Learning needs of nurses working in Canada's First Nations communities and hospitals. AB - BACKGROUND: What are the learning needs of nurses providing services to Canada's First Nations Communities and Hospitals? First Nations (or Indian Band) are similar to communities except some comprise more than one geographic Native community. Aboriginal (or Native) individuals are members of the North American Indian, Inuit, or Metis peoples of Canada; those who reported being a Treaty or Registered Indian (with the Federal Government); or those who are members of an Indian Band/First Nation. METHOD: A Canada-wide survey was completed to determine the learning needs of nurses working with Canada's Aboriginal persons. RESULTS: Nurses indicated both broad and specific aspects of their clinical practice, which were important to their continuing education (CE) needs. Broad thematic areas for continuing education included the following: emergency/acute care and obstetrics/gynecology clinical skills, health and physical assessment, mental health, and prenatal and postnatal care. Specific areas nurses cited for CE included issues related to: victims of violence; non-compliant clients; substance abuse; and fetal alcohol syndrome. CONCLUSION: This study examined the learning needs of nurses working with Canada's Native people and provided a basis for comparing and contrasting CE issues of these nurses to other nurses working in remote locations around the world. PMID- 11868712 TI - State and association/certifying boards: CE requirements. PMID- 11868714 TI - Our great need. PMID- 11868715 TI - The RN-BSN bridge course: transitioning the re-entry learner. AB - BACKGROUND: Although most RN-to-BSN programs offer bridge courses, little can be found in the literature about common course objectives, content, or outcomes. METHOD: This article details course objectives, course content, course implementation, and preliminary outcomes of a primarily distance learning RN-to BSN bridge course at a northern California university. RESULTS: Preliminary outcome assessments suggested greater than expected increases in role and campus socialization, as well as computer literacy, as a result of RN-to-BSN bridge course implementation. CONCLUSION: The RN-to-BSN bridge course can successfully be used to transition the re-entry student and may be the link allowing RN-to-BSN students to effectively face the concurrent challenges of role socialization and computer literacy while mastering course content and course objectives. PMID- 11868716 TI - Nurse responses to re-tooling practice, education, and management roles. AB - BACKGROUND: The purpose of this article is to report the results of a survey conducted by one of the nine Area Health Education Centers in North Carolina to inform regional work force planning by ascertaining nurses' views of the need to re-tool for practice, education, and management roles. METHOD: A work force planning group represented by nursing service and education surveyed 1,050 nurses. Four hundred thirty-two (41%) nurses responded: 87% believed they needed to re-tool and 92% were willing to do so. RESULTS: Critical thinking was rated as very important to nurse managers, staff RNs, and LPNs, and educators rated teaching as the area most in need of re-tooling. CONCLUSION: Nurses are clearly interested in re-tooling. Staff development, continuing education, and degree programs can respond to this willingness. PMID- 11868717 TI - Learner concerns and teaching strategies for video-conferencing. AB - BACKGROUND: The purpose of this study was to understand the influences of interactive video-conferencing technology on learning experiences of RN students studying for baccalaureate degrees via interactive distance education. METHOD: Data collection in this phenomenological study used open-ended questionnaires, interviews, and focus groups. Preliminary thematic analysis of questionnaires shaped open-ended questions for interviews and focus groups with learners confirmed findings. RESULTS: Students identified themes of connecting with others, organization, negative influences, and personal factors as influential to their learning. They also identified useful teaching strategies to facilitate learning within this distance nursing education environment. CONCLUSION: University nursing programs using video-conferencing for distance education can foster learning by using teaching strategies that fit the technology, increase student interaction, and engage the students. PMID- 11868718 TI - Learner support experienced by RNs in a collaborative distance RN-to-BSN program. AB - Sixteen RN-to-BSN students participated in interviews at the start and at the completion of taking their first class from a newly implemented, university based, collaborative distance learning program. The purpose of the taped, semi structured telephone interviews was to identify the support RNs experienced. Institutional themes of support that emerged were orientation programs, technical support, mentors, library resources, and prompt feedback from instructors. Non institutional support themes focused on family, friends, peers, the workplace, and learners. PMID- 11868719 TI - RN students' ratings and opinions related to the importance of certain clinical teacher behaviors. AB - Clinical teachers make a necessary and valuable contribution to clinical nursing courses with non-RN learners. This contribution is often not considered necessary in degree nursing programs with an RN student population. This study describes the perceptions of RN students and the importance they attach to certain clinical teacher behaviors. Fifty-six participants in a distance education baccalaureate nursing program completed a questionnaire, including a rating scale and open ended questions. Based on the mean values, items were ranked in importance. Sample quotes were provided to elaborate on the highest and lowest-ranked items. Participants rated the four categories in descending order of importance: Evaluation, Professional Competence, Interpersonal Relationships, and Teaching Ability. The top items gave the profile of a teacher who is approachable, fair, open, honest, and who creates mutual respect. The findings support a clinical teacher for RN learners. The data also reflect similarities and differences with findings of studies with non-RN student participants. The findings of this study provide information, not available in the literature, regarding clinical teacher behaviors of significance to RN students. PMID- 11868720 TI - Severity of illness--an important concept for critical care nursing? PMID- 11868727 TI - From unit to unit: Danish nurses' experiences of transfer of a small child to and from an intensive care unit. AB - In this study, Danish nurses' lived experience of transfer of a small child to and from the intensive care unit was explored. While there has been considerable research that has addressed transfer from the parents' perspective, little literature was found which addressed the transfer of small children from the nurses' perspective. A convenience sample of 19 nurses was interviewed once. Data were analysed following Spiegelberg's and Van Manen's phenomenological methodologies. Four themes emerged: being accountable; being supportive to the parents; being with the child; and experiencing safety and insecurity. Seven subthemes expanded and clarified the meaning of these themes. The study provides a thematic interpretation of how Danish nurses experience in-hospital transfers. Overall, the nurses were responsible to the transferred patient, the unhappy and worried parents, for technical procedures and the hospital team 'at home' on their own unit. However, responsibilities did not always include their colleagues on the receiving unit. It is recommended that transfer experiences be discussed more in clinical nursing, and that this explorative study needs to be followed by more studies exploring nurses' experiences of transfer. PMID- 11868728 TI - The meaning of social support for the critically ill patient. AB - Social support has been shown to be important for the critically ill patient. However, what constitutes adequate support for these patients has not been investigated. Thus, the purpose of this qualitative study was to investigate patients' perceptions of their need for and adequacy of the social support received while they were critically ill. Thirty adult patients who were critical during some point of their stay in the intensive care unit (ICU) stay were interviewed, once stable. Interviews were tape-recorded and began with an open ended question regarding the ICU experience. This was followed by open-ended focused questions regarding social support, such as 'Who were your greatest sources of social support while you were critically ill?' 'What did they do that was supportive or unsupportive?' Data were analyzed according to Miles and Huberman (1994). The categories that emerged were need for social support based on patient perceptions (not number of visitors), quality of support (based on perceptions of positive and negative behaviors of supporters) and lack of support. This study found that quality of support was more important than the actual number of visitors. Patients with few visitors may have felt supported, while those with numerous visitors felt unsupported. Patients who felt unsupported also were more critical of the staff and the care they received. Nurses need to individually assess patients regarding their need for support, and assist family/friends to meet these needs. PMID- 11868730 TI - Caregivers of ICU patients discharged home: what burden do they face? AB - Reforms in healthcare aimed at cost-cutting have led to early hospital discharge for many patients, including those who have sustained critical illness or injury requiring admission to an intensive care unit (ICU). Many former ICU patients experience ongoing health-related problems following discharge from hospital, and need ongoing support and care from their family once discharged. However, family members who take on a caregiving role often experience an enormous burden. The literature has demonstrated the impact of economic, social, physical and psychological factors on those who care for the critically ill following discharge. It is therefore essential that all nurses involved in the ongoing management of ICU patients have an understanding of the caregiving role, and consider both the patient and his/her carer in the discharge planning process. This paper reviews the literature on caregiving, and examines research studies which describe and measure the factors involved in the caregiving role. Although only a small number of studies specifically focus on caregivers of former ICU patients, findings suggest that these caregivers experience a substantial burden which is associated with the complexity of the patient's physical and psychological impairment, and complex technology. To enable nurses to incorporate planning strategies and interventions aimed at minimizing the impact on the caregiver, further research is needed to more fully explore, examine and measure the factors involved in caring for ICU patients post-discharge. Furthermore, ongoing research and evaluation of interventions will build an evidence-base to inform future practice. PMID- 11868729 TI - Nurses' knowledge and attitudes about antibiotic therapy in critical care. AB - PURPOSE: To assess critical care nurses' knowledge about antibiotic use in critical care settings, and attitudes concerning the role of the nurse in monitoring response to and appropriate use of antibiotic therapy. METHOD: 90 critical care nurses from 6 adult critical care units at a 780-bed academic, health sciences centre, completed an investigator-developed survey about their knowledge of antibiotic use and their attitudes concerning the role of the nurse. RESULTS: The majority of respondents worked full time (83%) and were BSN (Bachelor of Science in Nursing) prepared (62%), with an average of 9 years' nursing experience and 7 years' experience in intensive care. Using a 100-mm visual analog scale, mean scores on knowledge and comfort with: (1) interpreting culture and sensitivity; (2) white blood cell (WBC) data; and (3) discussing results and therapy with physicians were all less than 50 mm. However, the mean score for nurses' belief of responsibility related to this collaborative role was 76. A knowledge quiz of lab interpretation and antibiotic therapy revealed a mean score of 53.8%. Beliefs about roles were correlated with comfort in discussing therapies with physicians rather than with knowledge. Although nurses value the collaborative surveillance role, they may lack the knowledge and confidence to enact it. PMID- 11868731 TI - Dissonant imperatives in nursing: a conceptualization of stress in intensive care in Sweden. AB - The aim of this study was to explore nurses' experiences of stress within the context of intensive care. The theoretical perspective for the study builds on a cognitive-phenomenological-transactional theory of stress and coping and the theory of cognitive dissonance. Respondents were 36 registered nurses recruited from 10 intensive care units (general, neonatal and thoracic units). Their experience as nurses ranged from one to 32 years. These intensive care units had similar structural characteristics, namely a high working pace, advanced technology, constrained finances, frequent reorganizations, a shortage of registered nurses and all were filled to overcapacity. Data were collected in open-ended interviews that were audio-taped and transcribed. A content analysis identified four contradictory themes: (1) controlled by the work situation- needing to be in control; (2) constrained by prioritization--wanting to do more; (3) lacking the authority to act--knowing that something should be done; and (4) professional distance--interpersonal involvement. These four themes were synthesized at a higher level of abstraction into a main theme: stress induced by dissonant imperatives, which conceptualizes nursing stress in the intensive care unit. In conclusion, dissonant imperatives might lead to stress in intensive care nursing. PMID- 11868732 TI - Comprehensive critical care education is critical to success. AB - Intensive and high-dependency care is currently receiving considerable attention, not least since the Department of Health review of adult critical care nursing and subsequent proposals for the development of a consistent and comprehensive critical care service (DoH 2000). The report highlights the need for policy changes at all levels and across all disciplines and makes several recommendations that are deemed essential for the future organization and delivery of critical care. Central to the proposed reforms is the availability of a skilled workforce, adequately prepared and appropriately trained to provide specialist care in a variety of settings where acutely ill patients are found. This paper considers the implications of 'comprehensive critical care' for pre- and post-registration nurse education and offers insight into programmes of learning that are tailored to meet the needs of the service and individuals. The paper draws upon three influential and timely publications, namely the Audit Commission report into critical care services, the English National Board (ENB) review of adult intensive/critical and high-dependency care programmes and the United Kingdom Central Council (UKCC) Commission for Education recommendations regarding 'fitness for practice'. PMID- 11868733 TI - Steroids in sepsis--more effective than activated protein C? AB - Early studies using high doses of steroids in septic shock resulted in increased mortality. More recently, work by Annane et al. using lower doses closer to those that might constitute a stress dose of hydrocortisone have shown encouraging results with a 30% decrease in mortality in septic shock. Although this result is more impressive than the reduction in mortality resulting from the use of activated protein C in septic shock, the numbers of patients involved in the steroid study are low and the results remain unpublished in a peer-reviewed journal. PMID- 11868734 TI - Designing a conference--from start to finish. PMID- 11868735 TI - Continuing professional education, organizational support, and professional competence: dilemmas of rural nurses. PMID- 11868736 TI - Nurses' beliefs about their abilities to delegate within changing models of care. AB - Changing models of nursing care have resulted in a more diverse work force composition. Nurses (RNs and licensed practical and vocational nurses) have greater responsibilities for delegation and supervision of unlicensed assistive personnel providing direct nursing care. This study describes nurses' beliefs about their abilities to delegate and supervise direct nursing activities and explores differences based on professional and job-related factors. A national sample of 148 licensed nurses working in three practice settings was surveyed. In general, nurses reported a high level of comfort, frequency, preparedness, confidence, competence, and control. Differences found in nurses' beliefs were based on education, practice setting, and type of work responsibilities. PMID- 11868737 TI - Nutrition education for nursing assistants: an important strategy to improve long term care. PMID- 11868738 TI - Nutrition assessment: experience is not a predictor of knowledge. AB - This study tested an evidence-based nutrition education program for licensed nurses working in nursing homes who scored an average of 56% on a pretest. Posttest scores averaged 66%, demonstrating a continuing deficit of basic nutritional knowledge despite the educational intervention. Statistical analysis revealed that less experienced nurses scored better on posttests than more experienced nurses; more experienced nurses performed nutritional assessment more often than less experienced nurses; and more experienced nurses were usually charge nurses or nurses completing Minimum Data Sets. Although the education did not eliminate all deficits in nutrition knowledge, a decrease in the percentage of residents with significant weight loss was noted. PMID- 11868739 TI - The Pain Game: pain assessment, management, and related JCAHO standards. Joint Commission on Accreditation of Health Care Organizations. AB - BACKGROUND: It is a challenge to educators to assist nursing staff in maintaining up-to-date knowledge of pain assessment and management. METHOD: A learning game, "The Pain Game," was developed to deliver information on current practices and standards related to patient pain assessment and management. RESULTS: The advantages of using this gaming technique included minimal instructor preparation time, low cost materials, inclusion of participants as peer teachers, and a high level of participant involvement in the learning process. CONCLUSIONS: The Pain Game has been a well-received educational activity. Not only did knowledge acquisition occur, but the game also provided an enjoyable alternative to more traditional content delivery methods. PMID- 11868742 TI - Informal education: a hidden element of clinical nurse consultant practice. AB - A survey was conducted to investigate the amount of informal education provided by 16 community-based clinical nurse consultants (CNCs) in Sydney, Australia. The survey results indicated that the CNCs spent a substantial amount of time in informal teaching, of which only 3% was recorded as a legitimate educational activity. Findings from the survey support what has been reported in the literature, that skilled knowledge can be gained in the workplace when learners interact with experts. Through the avenue of informal teaching CNCs can positively contribute to quality nursing care by addressing on-the-job learning needs of community nurses. PMID- 11868740 TI - Fostering nursing research among nurse clinicians in an Australian area health service. AB - BACKGROUND: The Nursing Research Centre for Adaptation in Health and Illness (NRCAHI) is a collaborative effort of the University of Sydney and Central Sydney Area Health Service (CSAHS) in Sydney, Australia. This article reports findings of a research needs-analysis of clinical nursing staff in CSAHS that was conducted to provide research training. METHOD: A questionnaire was administered to a random sample of nurses working in CSAHS. The questionnaire gathered information about employment and demographic information, interest in research participation, current and future levels of research skills, and areas where skill and knowledge enhancement is desired. RESULTS: Overall, nurses expressed low levels of perceived confidence, knowledge, and research skills. Results were used as the basis for developing a range of strategies designed to enhance research confidence and participation. These strategies were sufficiently flexible to cater for the needs of nurses with varying levels of research experience. CONCLUSION: Strategies implemented by the NRCAHI in its first 4 years of operation have produced positive outcomes in terms of increased research participation. PMID- 11868743 TI - The effect of reading ability and response formats on patients' abilities to respond to a patient satisfaction scale. AB - This study investigated the relationship between reading ability and patient satisfaction measured with a 5-point Likert scale, a yes/no/uncertain choice, and a pictorial format. The study sample included 48 patients, 16 patients with third to fifth grade reading levels and 32 patients with reading levels of sixth grade or above. The mean percent agreement among the positively worded items on all three scales exceeded 80% regardless of reading level and response format. All of the patients had difficulty with the negatively worded items in all three formats, with the percent agreement dropping to only 49% between the negative items on the yes/no/uncertain and pictorial formats in the lower reading group. PMID- 11868744 TI - Continuing education for staff in long-term care facilities: corporate philosophies and approaches. AB - The purpose of this study was to determine corporate philosophies of continuing education and approaches to meeting the learning needs of staff who strive to provide for the increasingly challenging care requirements of seniors who reside in long-term care facilities. In-depth interviews lasting approximately 1 hour were conducted with key informants at the administrative level from nine long term care facilities. Content analysis revealed a commitment to continuing education for staff. While recognizing the importance of organizational responsibility for continuing education, administrators placed emphasis on the individual responsibility of staff. Learning needs were identified as affective, managerial, and physical in nature. Challenges to providing continuing education programs were derived from a general lack of fiscal and human resources. A variety of measures was suggested as important to supporting the continuing learning of staff. Implications of this study point to the need for long-term care facilities to incorporate into their strategic plans measures of ensuring continuing education as a basis for the ongoing competence and development of staff. In addition, there is a need for collaboration between long-term care facilities and other institutions of a long-term care, acute care, and educational nature in the development of strategies to operationalize a philosophy of continuing learning as a basis for the provision of optimal care to residents. PMID- 11868745 TI - Demonstrating theory in practice: examples of the McGill model of nursing. AB - An important test for a model of nursing is its usefulness in practice. One hundred and forty-seven nurses in one hospital attended a 2-day workshop on the McGill Model of Nursing. An evaluation of this workshop showed the usefulness of the McGill model. Self-efficacy for performing the model increased, as did behaviors and performance related to nursing by this model. Self-reports and supervisor reports measured these increases 6 months after the workshop. Further, the implementation of the model led to staff nurses initiating bottom-up changes in the nursing department. Boundary conditions or factors outside of the workshop content that enhanced the adoptation of the model are described. PMID- 11868747 TI - Nutritional status, delaying progression and risks associated with protein restriction. AB - Low protein diets (LPD), providing < or = 0.6 g protein/kg body weight daily, have traditionally been used in the nutritional management of chronic renal failure (CRF). Initially they were advocated when dialysis availability was limited. More recently LPD have been proposed as a means of delaying the rate of CRF progression. Despite much research in this area, the value of protein restriction remains contentious. A major concern over their use is that they may induce malnutrition. This is of critical importance since mortality rates are significantly increased in individuals who are malnourished when dialysis is initiated. Other approaches to the nutritional management of CRF may be more appropriate. The primary goal of any nutritional therapy should be to optimise the patient's nutritional status. In the UK LPD are used in a minority of units. An alternative approach to the use of these diets is to view the prevention of malnutrition as being of paramount importance. For uraemic patients, dialysis is the best treatment. PMID- 11868746 TI - Continuing education in Sweden--to what purpose? AB - BACKGROUND: Continuing education (CE) or recurrent education has long been used in Sweden to maintain and develop the competence of health care professionals. The purpose of this article is to describe the effects of CE with regards to changes in work activities, cognitive abilities, the use of knowledge, and involvement in research and development projects. METHOD: A questionnaire was distributed to all students (966 individuals) in CE courses at the Department of Advanced Nursing Education at Goteborg University, Sweden. RESULTS: Results indicated that CE leads to enhanced work satisfaction, extended work relevant competence, the attainment of professional goals, and enhanced self-confidence. The results further suggest that CE has not led to increased salaries, more power or authority, or greater possibilities to influence working situations. CONCLUSION: CE leads to positive development at the level of individual knowledge and competence but is not always reflected in changes in the respondents' working situations or in forms of rewards. The question is: How long are nurses prepared to continue raising their professional competence without compensation in terms of higher pay, better professional recognition, and new duties? PMID- 11868748 TI - Rehabilitating the frail and elderly on renal replacement therapy. AB - The World Federation of Occupational Therapists defines Occupational Therapy as "...the treatment of physical and psychiatric conditions through specific activities in order to help people reach their maximum level of function and independence in all aspects of daily life." Occupational Therapy has long been associated with the frail and elderly, both in hospital and the community. In the United Kingdom however, it appears formal input to the renal multidisciplinary team is a relatively new development. Certainly across Europe, none of us needs reminding of the increasing incidence of older patients with renal failure. United Kingdom figures released in the Second Annual Report of the UK Renal Registry in December 1999 state that, "At the start of treatment 46% of patients were aged 65 or more" (1). PMID- 11868749 TI - Advanced nursing practice in renal medicine. AB - Renal Nursing presents many challenges for nurses seeking to work at advanced level. The ever-changing political climate and increasing service demands make it vital for nurses to grasp the opportunities that advanced practice offers. Theorists have suggested that the over-specialisation of nursing could lead to a fragmented nursing service and an undervaluing of the registered nurse. September 1999 saw the appointment of two such Advanced Nurse Practitioners within the renal unit. This article takes a personal look at the implementation of an Advanced Nurse Practitioner service over the first 12 months and suggests that such a role does have a lot to offer particularly in a renal setting. PMID- 11868751 TI - The pre-dialysis experience--are individual needs being met? AB - The pre-dialysis experience is a time of change. Individuals and their families are able to adapt to this change and altered lifestyle if their specific needs are met. This qualitative study aims to gain an understanding of the patient's. perspective and identify how they manage the pre-dialysis experience. This knowledge will enable renal unit teams to plan effective holistic programmes, which meet the needs of the individual. The emergent theory identified that pre dialysis patient's and their families have to accept a different view of life before they can move forward and make a decision regarding treatment. The adoption of a model for good practice aids the transition. PMID- 11868750 TI - Improvement in care: a collaborative approach to rehabilitation. AB - Examination of the patient experience within our unit, from pre dialysis through to establishment of dialysis in the community identified that the care was fragmented. To improve patient care, a change process was initiated. Four home care teams comprising three qualified nurses and one renal care assistant were established with each team responsible for a caseload of patients within a specified geographical location. To measure the impact on the patient, 100 questionnaires were circulated after twelve months. Results from 60 patients showed 76% of pre dialysis patients and 80% of dialysis patients were very satisfied with the change process. The main advantage of this change for the patient is that they are in a continuous supportive cycle for all their non inpatient care throughout their replacement therapy. We conclude that patient focused care is essential and should be a transition catalyst in a change resistant environment. PMID- 11868752 TI - Clinical impact of residual renal function on patients starting dialysis treatment. AB - We describe our clinical experience of monitoring residual renal function of patients on regular dialysis treatment as residual renal function can play an important role in the management of patients undergoing regular dialysis. With active residual renal function it is possible to delay the start of haemodialytic programmes. In managing dialysis treatments, it is important to maintain residual renal function by preventing hypotensive events. It is known that in acute renal failure intradialytic hypotension is able to reduce the recovery of renal function. Out patient follow-up plays an important role for these patients. This paper will also discuss the use of biocompatible membranes during dialysis. The outcome is that they are not associated with better maintenance of residual renal function. PMID- 11868753 TI - 21st century home haemodialysis: a new approach to an old treatment. AB - Home therapies are increasingly being demonstrated to be the best treatments for the early stages of the dialysis life-cycle. Although home haemodialysis has declined dramatically over the last 20 years from 41% in 1983 (1) to 3.2% in 1998 in the UK alone (2), many studies have suggested that it offers the optimum dialysis in terms of outcomes (3,4,5). Evidence from a 1998 survey of UK dialysis staff indicates that the major perceived drawbacks of home haemodialysis were lack of suitable patients, family stress, cost of machines and training time (6). The study also strongly indicated that a lack of familiarity with the treatment was a major cause of its decline in many units. We set out to redesign our approach to home haemodialysis to make it suitable for many more patients. PMID- 11868754 TI - Maintaining an optimum haemocatheter exit site. AB - As a result of crusty growths over some haemocatheter exit sites, it was decided to explore why these crusty scabs occur and how they could be eradicated. It is concluded that a haemocatheter exit site requires a dressing, which will maintain an optimum environment, and avoid contamination from Staphylococcus aureus. Inadine (Johnson & Johnson) a povidine iodine product, applied before the final dressing of Tielle (Johnson & Johnson Medical), a hydropolymer adhesive dressing creates an ideal environment for healing. The haemocatheter exit site can also be inspected at each dialysis session, but can in fact be left insitu for seven days. PMID- 11868755 TI - Percutaneous interventions for vascular dialysis access. AB - A prerequisite for the long-term patency of vascular access is early detection of a failing shunt, i.e., at the stage of stenosis. Fistulography performed to locate the lesion is followed by percutaneous transluminal angioplasty using a balloon catheter. The procedure takes 30-60 minutes to complete; the clinical success rate varies between 85% and 98%. A more serious condition of failled vascular access is its occlusion secondary to thrombosis. The high costs of hospitalisation and treatment of thrombosed shunts require effective yet less invasive outpatient procedures. Recent years have seen an explosion in the development of mechanical devices on the principle of mechanical dissolution, fragmentation, and aspiration of the clot. Post-procedural dialysis can be performed immediately thus reducing the need for a central venous catheter. The procedure, particularly in the treatment of native fistula occlusion, is more complicated and about twice as time consuming as treatment of stenosis. The clinical success rate is usually between 71% and 100%. The incidence of serious complications of percutaneous treatment is low. PMID- 11868757 TI - Japanese patients not seeking kidney transplants. AB - In Japan, the number of patients seeking kidney transplants has declined in recent years. To investigate the reasons for this decline, a questionnaire was given to 73 haemodialysis patients treated at the Tokiwa-Tatsumi Clinic. The resulting data showed the percentage of patients seeking transplants declined from 61% in 1992 to 19.2% in 1999. The reasons given for not seeking transplants were the improvements of physical condition and resultant quality of life (QOL) due to progress in dialysis therapy, upgraded social welfare support, uncertainties of transplant medicine, loss of expectations due to limited availability of transplant kidneys and aging of patients. Meanwhile, the number of patients on dialysis continues to increase by approximately 10,000 a year, and the mean age of patients rises. To reduce this number, greater effort must be directed toward preventive medical care as well as educating the public regarding transplant medicine. PMID- 11868756 TI - Drop-out rate during living donor selection. AB - This unit carried out 29 live donor transplants over a 3-year period. Many potential donors did not proceed to transplant. For those who had an acceptable tissue type, were blood group compatible and lymphocytotoxic crossmatch negative, we looked at the reasons for cancelling the donor work up. The reasons were impaired renal function (5 potential donors), cardiac/hypertension (4 potential donors), renovascular (1 potential donor), cancer (1 potential donor), cross match positive at a late stage (3 potential donors), failure to attend at clinic/change of mind (6 potential donors) and hepatitis (2 potential donors). Improvements carried out following the audit include a list of tests which potential donors living away from this unit--especially those abroad--are asked to do before travelling here. An information leaflet has also been produced for potential donors. PMID- 11868758 TI - The UK's Asian population: solving the transplant crisis. AB - The United Kingdom Transplant Authority has recently re-introduced a policy to identify the ethnicity of patients. This is in response to the realisation byTransplant Co-ordinators and other health professionals in the field that a number of disparities existed between Asian residents and the indigenous population. The limited data that exists highlights that the Asian population are in a disproportionately greater need of kidney transplants. The situation is clear, there needs to be a greater number of donors coming forward from the Asian communities to increase the pool of suitable organs. However, this may only be achieved if we understand the reasons for the current lack of supply. Very little empirical research has been devoted to this subject and those studies, which have been carried out, highlight the need for greater attention to this life threatening problem. PMID- 11868759 TI - Efficacy of short-course therapy with the ketolide telithromycin compared with 10 days of penicillin V for the treatment of pharyngitis/tonsillitis. AB - This randomized, double-blind study compared the efficacy and safety of a 5-d course of the new ketolide antimicrobial, telithromycin, with those of a standard 10-d course of penicillin V (phenoxymethylpenicillin) in patients with group A beta-hemolytic streptococci (GABHS) pharyngitis/tonsillitis. Patients aged 15-65 y (n = 395) with clinical signs and symptoms of pharyngitis/tonsillitis and a positive streptococcal antigen test or throat culture for GABHS were randomized to receive either telithromycin 800 mg once daily for 5 d (n = 198) or penicillin V 500 mg three times daily for 10 d (n = 197). Clinical and bacteriologic outcomes were assessed at post-therapy, test-of-cure (Days 16-20) and late post therapy (Days 38-45) visits. Telithromycin for 5 d was equivalent to 10 d of penicillin V in terms of bacteriologic and clinical outcome (per-protocol): at post-therapy, test-of-cure visit, bacteriologic outcome was satisfactory in 84.3% and 89.1% of patients in the telithromycin and penicillin V groups, respectively, while clinical cure was achieved in 94.8% and 94.1% of patients, respectively. At late post-therapy, 82.4% of patients treated with telithromycin achieved a satisfactory bacteriologic outcome, compared with 84.7% of penicillin V recipients. The GABHS eradication rates for telithromycin and penicillin post therapy were 85.2% and 89.1%, respectively, and 86.1% and 86.5%, respectively at late post-therapy. Both treatments were well tolerated, with a similar overall incidence of treatment-emergent adverse events. Short-course (5 d) therapy with telithromycin 800 mg once daily is comparable to a standard 10 d course of penicillin V for the treatment of GABHS pharyngitis/tonsillitis in adults and adolescents. PMID- 11868760 TI - Prospective study of antibacterial susceptibility, risk factors and outcome of 157 episodes of Acinetobacter baumannii bacteremia in 1999 in Slovakia. AB - This study prospectively investigated all 157 cases of Acinetobacter baumannii bacteremia occurring in major university hospitals or tertiary care institutions in Slovakia during 1999 in order to determine the antimicrobial susceptibility, risk factors and outcome. Resistance to meropenem was 7.4, gentamicin 35.6, amikacin 26.5, cefepime 20.4 and ciprofloxacin 32.7%, but was only 17.3% to cefoperazone/sulbactam or ampicillin/sulbactam. Antimicrobial susceptibility of A. baumanii was lowest among isolates from cancer patients (ceftazidime 58%, piperacillin/tazobactam 52% and azthreonam 48%; p < or = 0.01-0.001). In univariate analysis, several risk factors, such as wound infection (p < or = 0.01) and ventilatory support (p < or = 0.0001), were significantly related to A. baumannii bacteremia in surgical patients. Neutropenia (p < or = 0.0001), antineoplastic chemotherapy (p < or = 0.0001) and prior antibiotic therapy (p < or = 0.0006) were significant risk factors for A. baumannii bacteremia in cancer patients. In addition, ventilatory support and surgery (p < or = 0.0001) and prior antibiotic therapy (p < or = 0.01) were significantly related to A. baumannii bacteremia in children. Colonization at other body sites (p < or = 0.05), diabetes mellitus (p < or = 0.04) and decubital ulcers/burns (p < or = 0.002) as underlying disease were significantly related to death due to A. baumannii bacteremia. In a multiple logistic regression model, decubital ulcers/burns as underlying disease (p < or = 0.0006; relative risk 5.08) and nosocomial pneumonia (p < or = 0.045; relative risk 5.08) were independent predictors of mortality. Mortality was similar between cancer and surgical patients but significantly lower in children vs. adults (p < or = 0.009). PMID- 11868761 TI - Rattus norvegicus acting as reservoir of leptospira interrogans in the Middle Black Sea region of Turkey, as evidenced by PCR and presence of serum antibodies to Leptospira strain. AB - Leptospirosis, a common form of zoonosis, especially in rainy countries, is caused by Leptospira interrogans. In our region of Turkey this type of disease has often been encountered in connection with rice harvesting and we therefore attempted to evaluate the prevalence of L. interrogans in wild rats in our region. Fifty-nine Rattus norvegicus rats were trapped alive in different areas of an approximately 100 km stretch of seashore in the Middle Black Sea region of Turkey. L. interrogans was determined by PCR in sera, kidney and brain tissue. Sixteen (27.1%) kidney samples and 10 brain tissue samples (16.9%) were positive for L. interrogans. No PCR positivity was seen in sera samples. Five sera were positive by microagglutination test. A large proportion of wild rats in our region were found to be carriers of L. interrogans. We conclude that people who are exposed to rat urine in their daily life are at risk of acquiring L. interrogans. PMID- 11868762 TI - Ecological effects of linezolid versus amoxicillin/clavulanic acid on the normal intestinal microflora. AB - Twelve healthy subjects (6 females, 6 males; age range 18-40 y) participated in this trial. Linezolid was given as 600 mg tablets b.i.d. for 7 d and amoxicillin/clavulanic acid as 1000 mg tablets o.d. for 7 d. The washout period between the administration of the 2 antibacterial agents was 4 weeks. Faecal samples were collected prior to administration (Days -2 and -1), during administration (Days 4 and 8) and after administration (Days 14, 21 and 35) for microbiological analyses. The samples were diluted in pre-reduced media and inoculated aerobically and anaerobically on non-selective and selective media. Different colony types were identified to genus level by morphological, biochemical and molecular analyses. During the administration of linezolid, enterococci in the intestinal aerobic microflora were markedly suppressed while Klebsiella organisms increased in number. In the anaerobic microflora, the numbers of bifidobacteria, lactobacilli, clostridia and Bacteroides decreased markedly while no impact on the other anaerobic bacteria was observed. The microflora was normalized in all volunteers after 35 d. Amoxicillin/clavulanic acid administration caused increased numbers of enterococci and Escherichia coli in the aerobic intestinal microflora while numbers of bifidobacteria, lactobacilli and clostridia decreased significantly. Clostridium difficile strains were recovered from 3 of the volunteers. At the last visit, the intestinal microflora of the volunteers had returned to normal levels. The administration of linezolid mainly had an impact on the gram-positive bacteria and linezolid thus had an ecological profile different from that of amoxicillin/clavulanic acid. PMID- 11868763 TI - Impact of IgG replacement therapy and antibiotic treatment on the colonization of non-encapsulated Haemophilus influenzae in the nasopharynx in patients with hypogammaglobulinaemia. AB - Non-typable Haemophilus influenzae (NTHI) and Streptococcus pneumoniae are regarded as the main pathogens in patients with humoral immunodeficiency. These patients have been given IgG replacement therapy since the 1950s. However, a number of individuals still suffer from recurrent episodes of respiratory tract infections. Nasopharyngeal cultures were obtained on a regular basis over a 3-6 month period from 11 patients with common variable immunodeficiency disease in 1989 and 1998. The proportion of cultures positive for NTHI decreased from 56% in 1989-90 to 16% in 1998-99 (p < 0.003). After 9 y of IgG therapy, 7 of the 11 patients were free from NTHI in the nasopharynx. Specific NTHI strains were analysed by pulsed-field gel electrophoresis and compared, with regard to genetic relatedness, on an intra-individual basis. In 1 patient the same NTHI strain was found in both 1989 and 1999. The apparent absence of NTHI from the nasopharynx in most of the patients was assumed to be due to subcutaneous IgG treatment, as well as adequate antibiotic treatment. PMID- 11868764 TI - D- and L-lactate in rabbit and human bacterial meningitis. AB - Increased total CSF lactate is an important indicator differentiating bacterial from aseptic meningitis. Bacteria can produce D- and L-lactate; mammalian cells produce only L-lactate. We measured D- and L-lactate production of Streptococcus pneumoniae, Staphylococcus aureus, Neisseria meningitidis and Escherichia coli in vitro, of S. pneumoniae and E. coli in rabbit experimental meningitis and of various common pathogens in CSF from patients with bacterial meningitis. Despite marked in vitro production of D-lactate by S. aureus (maximum: 4.59 mmol/l; i.e. 34.9% of total lactate), N. meningitidis (4.62 mmol/l; i.e. 98.1%) and E. coli (3.14 mmol/l; i.e. 97.2%), minimal amounts were measured in human S. aureus (0.38 mmol/l; i.e. 1.3% of total lactate) or N. meningitidis (0.28 mmol/l; i.e. 3.9%) and experimental E. coli meningitis (0.75 mmol/l; i.e. 4.4%). In only 9 of 54 human CSF samples did D-lactate exceed 0.15 mmol/l. S. pneumoniae did not produce significant amounts of D-lactate in vitro (maximum: 0.55 mmol/l; i.e. 2.7% of total lactate), in experimental meningitis (0.18 mmol/l; i.e. 3%) or in human cases of meningitis (0.28 mmol/l; i.e. 1.9%). In conclusion, increased total CSF lactate in meningitis consists mainly of L-lactate and originates predominantly from host cells. CSF D-lactate is of limited diagnostic value. PMID- 11868765 TI - Sensitivity to anti-tuberculosis drugs in HIV-positive and -negative patients in Addis Ababa. AB - A cross-sectional survey was conducted between 1 August and 31 December, 1998 in Addis Ababa, Ethiopia to determine the rate of primary drug resistance to anti tuberculosis drugs and to investigate its possible association with HIV infection. Sputum culture, sensitivity to first-line anti-tuberculosis drugs and HIV testing were done for 236 sputum smear-positive pulmonary tuberculosis patients. Primary drug resistance level (single or multidrug resistance) had not changed significantly since 1994. Primary resistance occurred more often among HIV-positive than among HIV-negative patients. The association between drug resistance and HIV will have a serious impact in the control of tuberculosis because in recent years the prevalence of HIV has increased dramatically in Ethiopia. Therefore, further studies on drug resistance and HIV infection and the establishment of drug resistance surveillance are recommended. PMID- 11868767 TI - Recurrent septicemia caused by Streptococcus canis after a dog bite. AB - Human infection with Streptococcus canis is extremely rare. We describe herein a case of septicemia with cellulitis caused by S. canis in a 75-y-old woman, which developed 2 weeks after a dog bite. Macrorestriction analysis with pulsed-field gel electrophoresis demonstrated that the organism had been transmitted by means of a dog bite to her hand. PMID- 11868766 TI - Antibiotic use by indication: a basis for active antibiotic policy in the community. AB - The aim of this study was to survey current treatment practices for common infections in primary care as a basis for implementation of recently released evidence-based guidelines for community-acquired infections. A point-prevalence survey was conducted in 30 health centres in the Finnish primary care system with a population base of 819,777. All patients consulting the health centres for an infection during a 1-week period were included in the study. The main outcome measures were the prevalence of antibiotic prescription and the selection of drugs by infection diagnosis. Of the 7777 recorded consultations, 85% were with a physician and the rest with a nurse. The most common cause for a visit was respiratory tract infections (74%), followed by skin/wound infections and urinary tract infections (both 6%). The infection panorama varied markedly according to age: in the youngest children (< 5 y) 84% of the infections were respiratory tract infections whereas the corresponding figure for patients > 65 y was 50%; the proportions of visits for urinary tract infections in these age groups were 7% and 26%, respectively. Of the patients with acute bronchitis, 70% were treated with antimicrobial agents, mostly macrolides (39%) and doxycycline (36%). Of the otitis media patients, 53% were treated with amoxicillin, 16% with macrolides and 16% with sulphatrimethoprim. Macrolides were mostly used to treat otitis media (31%), acute bronchitis (26%) and sinusitis (20%). In conclusion, antimicrobial agents are still used excessively in Finland, particularly for the treatment of acute bronchitis. Moreover, the selection of drugs for treating sinusitis and otitis media is non-optimal; macrolides and cephalosporins are frequently chosen unnecessarily. Knowledge of the indication-based prescription practices for antimicrobial agents is essential in order to improve the treatment habits of primary care physicians. The data obtained in this study provide a unique tool for the active and targeted implementation of evidence-based guidelines for primary care physicians. PMID- 11868768 TI - Meningococcemia following tonsillectomy. AB - A 22-y-old male developed fever, arthralgias and skin rash 2 d after tonsillectomy. Blood culture grew Neisseria meningitidis. He responded well to antibiotics. Only 1 other case of meningococcal sepsis following adenotonsillectomy has previously been reported. These cases suggest a temporal association of meningococcemia with tonsillectomy. PMID- 11868769 TI - Intravenous catheter-related septic shock caused by Staphylococcus sciuri and Escherichia vulneris. AB - Staphylococcus sciuri and Escherichia vulneris were isolated concurrently in a blood sample from a patient with septic shock, which was probably associated with an indwelling catheter. Our results also showed that S. sciuri is an important reservoir of genetic determinants of beta-lactam resistance as a human pathogen carrying the mecA and beta-lactamase genes. PMID- 11868770 TI - Myroides odoratus cellulitis and bacteremia: case report and review. AB - A case of Myroides odoratus cellulitis with bacteremia in an apparently immunocompetent man is presented. Although common in soil and water, this agent is a rare clinical isolate and is often not considered pathogenic. The virulence of M. odoratus may be greater than is currently believed and it should be considered in bacteremias from cutaneous sources in immunocompetent patients. PMID- 11868771 TI - Bartonella henselae infection mimicking a splenic lymphoma. AB - We report a Bartonella henselae infection in a 40-y-old patient who presented with fever, weight loss, night sweats, elevated lactate dehydrogenase and multinodular splenomegaly with multiple abdominal lymphadenopathies. Splenic cat scratch disease is an exceptional diagnosis in adults and can easily be mistaken for a splenic lymphoma, thereby leading to an unnecessary splenectomy. PMID- 11868772 TI - Mycobacterium avium complex peritonitis in an AIDS patient. AB - Mycobacterium avium complex (MAC) frequently disseminates in AIDS patients, where the gastrointestinal tract is a major target organ. While ascites in AIDS patients is common, peritonitis secondary to MAC is rare. We describe the first case of MAC peritonitis in an AIDS patient without underlying cirrhosis, portal hypertension, chylous ascites or peritoneal dialysis. This case highlights the need to be aware of atypical presentations of MAC disease in AIDS patients with a history of disseminated MAC, even those who compliantly take highly active antiretroviral therapy. PMID- 11868773 TI - Antiphospholipid syndrome associated with intestinal amoebiasis. AB - We present a case of intestinal amoebiasis with subsequent development of antiphospholipid syndrome, manifested by deep vein thrombosis and pulmonary emboli. Anticardiolipin antibodies (aCL) of IgM type at medium titer and aCL IgG antibody at low titer were determined during the days after the onset of infection. To our knowledge this is the first case of antiphospholipid syndrome associated with amoebiasis to be presented in the literature. PMID- 11868774 TI - Repeated unintentional transthoracic needle aspiration of a pulmonary hydatid cyst. AB - Transthoracic needle aspiration is not a recommended diagnostic modality in hydatid disease. Percutaneous aspiration of a suspected hydatid cyst is believed to be associated with the risk of allergic reactions which can result in systemic anaphylaxis and possible spreading of the cyst contents. We present herein a illustrative case of pulmonary hydatid cyst and multiple mediastinal lymphadenopathies which was diagnosed after repeated transthoracic fine needle aspirations. PMID- 11868775 TI - Fatal human herpesvirus 6-associated multifocal meningoencephalitis in an adult female patient. AB - Human herpesvirus 6 (HHV 6) is a known cause of central nervous system infection in immunocompromised patients. Less is known about the clinical course of HHV 6 encephalitis in immunocompetent patients. We report a case of meningoencephalitis in a 42-y-old immunocompetent patient associated with HHV 6 infection. PMID- 11868776 TI - Cutaneous tuberculosis from paradoxical expansion: a new category in the classification of cutaneous tuberculosis. PMID- 11868778 TI - A message to Asia. PMID- 11868777 TI - A message to Japan. PMID- 11868779 TI - Natural history of hepatitis B and C virus infections. AB - Thirty-five years have already elapsed since the discovery of hepatitis B virus (HBV), and 10 years since that of hepatitis C virus (HCV). Nonetheless, the natural history of HBV and HCV infections has not been fully defined, partly because they do not have subjective symptoms in most cases. Even when liver disease is induced by these hepatitis viruses, the clinical course is slow and mostly insidious. HBV and HCV are much alike in that they both cause a spectrum of clinical conditions ranging from the symptom-free carrier state through chronic hepatitis and liver cirrhosis to eventual hepatocellular carcinoma. Despite a close similarity, infections with HBV and HCV are very different in many aspects, from the early to end stage. Large-scale unbiased studies to sort out the natural history of HBV and HCV infections are lacking, however. Understandably, in view of the fact that only a few decades have passed since these hepatitis viruses were discovered. My personal account on the natural history of HBV and HCV infections is given here, which is based on my experience of over 40 years as a clinical and research hepatologist in Japan, although I am aware that it invites more questions than it answers. PMID- 11868780 TI - Leading-edge research studies of hepatocellular carcinoma in Japan. PMID- 11868782 TI - Molecular mechanism of viral hepatocarcinogenesis. AB - Overwhelming lines of epidemiological evidence have indicated that chronic infection with hepatitis B virus (HBV) or hepatitis C virus (HCV) is a major risk for the development of hepatocellular carcinoma (HCC). In the pathogenesis of HCC associated with HBV or HCV, it remains controversial whether these hepatitis viruses play a direct role or merely an indirect role. By virtue of transgenic mice established by us, it has become evident that the product of the HBV X gene (HBx protein) and the core protein of HCV have an oncogenic potential, although the pathways through which these two viral proteins operate may differ. The findings in our studies indicate that HBV and HCV are directly involved in hepatocarcinogenesis, albeit other factors such as continued cell death and regeneration associated with chronic hepatitis may play a role as well. Combined, our results suggest that there might be a mechanism in the development of HCC in persistent infection with hepatitis viruses that is distinct from that in other cancers. Similarly to the pathogenesis of other malignancies represented by colorectal cancer, the accumulation of a set of genetic aberrations may also be necessary for a multistage development of HCC. However, HBx protein and HCV core protein, to which an oncogenic potential is attributed, may allow some of the multiple stages skipped in hepatocarcinogenesis. Unlike for the other cancers, therefore, infection with HBV or HCV may be capable of inducing HCC in the absence of a complete set of genetic aberrations. Such a scenario would explain an unusually high incidence and multicentric nature of HCC developing in chronic hepatitis B or C. PMID- 11868781 TI - Current strategies for chemoprevention of hepatocellular carcinoma. AB - Hepatocellular carcinoma (HCC) develops frequently in the liver of patients with chronic hepatitis and cirrhosis caused by persistent infection with hepatitis B virus or hepatitis C virus in Asia. Several studies including ours have revealed that the risk of HCC increases in parallel with the progression of hepatic fibrosis associated with chronic hepatitis and liver cirrhosis. It may be delineated reasonably, therefore, that suppression of fibrosis by chemical (or biological) agents would be able to decrease the risk of HCC. We have demonstrated that antifibrotic agents, such as HOE 077, TJ-9 and interferon, can reduce the risk of HCC. Accordingly, antifibrotic agents are promising candidates for chemoprevention of HCC. The results of our study are discussed within the scope of the current state of the art as regards chemopreventing HCC. PMID- 11868783 TI - Molecular aspects of human hepatocarcinogenesis mediated by inflammation: from hypercarcinogenic state to normo- or hypocarcinogenic state. AB - In spite of great efforts in the research relating to human hepatocarcinogenesis, its mechanism on the molecular level is yet to be determined. Chronic viral hepatitis may increase the chances of genetic events in hepatocytes of the host, by increasing the number of target cells or through proliferation of initiated hepatocytes, toward the eventual development of hepatocellular carcinoma. These conditions are referred to comprehensively as the 'hypercarcinogenic state'. Our goal, then, should be directed to the reversion of the 'hypercarcinogenic state' to the 'normo- or hypocarcinogenic state' so as to hopefully prevent or at least postpone the development of hepatocellular carcinoma. PMID- 11868784 TI - Pathological evolution of early hepatocellular carcinoma. AB - Recent remarkable advances in the study of tumorigenesis for hepatocellular carcinoma have implicated a pivotal role of dedifferentiation, which is supported by morphology and experimental evidence. Understanding the mechanism of how hepatocellular carcinoma develops would help plan strategies for a better prognosis for the patients suffering from it. PMID- 11868785 TI - Imaging blood flow characteristics of hepatocellular carcinoma. AB - Since a close relationship exists between intranodular hemodynamics and the grade of biological/pathological malignancy of a nodule occurring in the cirrhotic liver, an accurate evaluation of intranodular hemodynamics is highly essential. Intranodular hemodynamics in hepatocellular carcinoma (HCC) and borderline lesions can be evaluated correctly by invasive and noninvasive techniques. Invasive techniques such as ultrasound (US) angiography, computed tomographies during arteriography or arterial portography are sensitive in the detection of intranodular arterial and portal supplies, for accurate diagnosis of tumors and assessing grades of biologically malignant potential. However, these approaches require an angiographic procedure, which is not always available. Recently, perfusion imaging techniques under US, including contrast-enhanced harmonic imaging or real-time gray-scale harmonic imaging, have become available for routine clinical use. With these techniques, all the five roles of imaging in the management of HCC, i.e., detection, confirmation, staging, evaluation of malignancy grade, and postoperative follow-up, have become much simpler. Perfusion imaging techniques have reduced the requirement for dynamic CT or MRI and may replace some of their roles in the clinical setting. Since viable cancer cells are accurately imaged on US monitoring with sensitive perfusion imaging techniques, the contrast-enhanced harmonic imaging will be of great advantage in US-guided treatment of HCC. With the advent of rapid and remarkable advances in US harmonic imaging techniques, the diagnostic and therapeutic strategies for HCC are changing drastically. PMID- 11868786 TI - Characteristics of hepatocellular carcinoma in Japan. AB - Malignant neoplasias are the leading cause of death in Japan at present, of which hepatocellular carcinoma (HCC) ranks the third most frequent in men and the fourth most frequent in women. Annual deaths due to HCC are rising sharply and amounted to 33,000 in 1999. Chronic hepatitis C is the most frequent etiology of HCC in Japan and accounts for nearly 90% of cases. The recent rapid increase of HCC in Japan is a long-term sequel of hepatitis C virus infection that affected many individuals in the past, and manifests itself currently as HCC. Infection with the hepatitis C virus has prevailed since the end of World War II for reasons inherent in the socioeconomic background in Japan. PMID- 11868787 TI - Tumor markers in early diagnosis, follow-up and management of patients with hepatocellular carcinoma. AB - The mainstay for the diagnosis for hepatocellular carcinoma (HCC) includes serological tumor markers, such as alpha-fetoprotein, the L3 fraction thereof and PIVKA-II, in addition to imaging modalities. They do not correlate, but complement each other. Hence, a combination of them designed on the basis of their characteristics needs to be worked out. First, it is necessary to identify the patients at high risk for developing HCC, such as those with chronic hepatitis or liver cirrhosis, and in the follow-up conduct regular check-ups for serological tumor markers. Those testing positive for any marker are at the highest risk for developing HCC, even when imaging fails to disclose any space occupying lesions. Following high-risk patients for serological tumor markers, in concert with imaging, makes accurate evaluation of the efficacy of therapies for HCC possible. Since serological tumor markers can signal the development of HCC earlier than any other laboratory tests, they offer excellent means of identifying relapsing HCC. Equally important in the management of patients with HCC are biological indicators for malignancy, the selection of therapeutic interventions and the prediction of the outcome. PMID- 11868788 TI - Nonsurgical treatment of hepatocellular carcinoma: from percutaneous ethanol injection therapy and percutaneous microwave coagulation therapy to radiofrequency ablation. AB - Treatment of hepatocellular carcinoma (HCC) is different from that of other solid tumors, in that surgery plays a limited role while nonsurgical therapies are very instrumental. At our institute, 90% of previously untreated patients have received image-guided percutaneous tumor ablations, such as percutaneous ethanol injection therapy (PEIT), percutaneous microwave coagulation therapy (PMCT) and radiofrequency ablation (RFA). We performed PEIT in 756 patients with HCC. Their survival rates were 89% at 1 year, 64% at 3 years, 39% at 5 years, and 18% at 10 years. With PMCT, survival rates of 122 new patients with HCC were 90% at 1 year, 87% at 2 years, and 68% at 3 years. We performed RFA in 324 patients. RFA required fewer treatment sessions and a shorter hospital stay than PEIT or PMCT to achieve complete necrosis of the lesions. By virtue of their local curability, minimal effect on liver function, and easy repeatability for recurrence, image guided percutaneous tumor ablations, especially RFA, will be increasingly important in the treatment of HCC. PMID- 11868789 TI - Combination chemotherapy for advanced hepatocellular carcinoma complicated by major portal vein thrombosis. AB - Patients with advanced hepatocellular carcinoma (HCC) have a poor prognosis, and the development of new therapeutic strategies is necessary. Here we report the efficacy of combination chemotherapy in our biochemical modulation. Synergistic effects of interferon-alpha-2b on 5-fluorouracil or cisplatin were demonstrated in Huh7 cells. The efficacy of methotrexate-5-fluorouracil, cisplatin, and interferon-alpha-2b combination therapy was demonstrated in 34 patients with HCC complicated by major portal vein thrombosis. Among the 29 patients eligible for the study, there were 3 complete responders and 10 partial responders with an overall response rate of 45%. The 2-year survival of the 34 patients was 15%, and the median survival of complete and partial responders was 11 months. There was severe transient hematological toxicity. Eight patients suffered from renal insufficiency, and 4 of them underwent hemodialysis. Although a control study is clearly necessary, our combination therapy induced a good response in the patients with advanced HCC. On the basis of our results, intensive chemotherapy should be attempted in advanced HCC complicated by major portal vein invasion. PMID- 11868790 TI - Progress in surgical treatment of hepatocellular carcinoma. AB - Surgery for hepatocellular carcinoma has improved dramatically during the last two decades, and the improvement is mainly attributable to the development of intraoperative ultrasound-guided operative procedures such as Makuuchi's segmentectomy, introduction of the intermittent vascular occlusion technique, and establishment of the precise criteria for indications of various hepatectomy procedures. The use of preoperative portal vein embolization for inducing compensatory hypertrophy of remnant liver in the future has increased the safety and extended indications of hepatectomy for hepatocellular carcinoma. Operative mortality has fallen below 2% in the 1990s, with the 5-year survival rate reaching nearly 50% in a recent nationwide survey in Japan. More than 90% of hepatectomies at our institution are performed without red blood cell transfusions, and the mean hospital stay is shortened to approximately 23 days. Moreover, not a single case of operative death has been recorded since 1993. PMID- 11868791 TI - Hepatocellular carcinoma associated with hepatitis C virus infection in Japan: projection to other countries in the foreseeable future. AB - During the turmoil after the end of World War II some 50 years ago, in Japan intravenous methamphetamine was widespread and penetrated the young generation aged 15-25 years and remunerated blood donors. The vicious cycle gave an enormous thrust to the spread of hepatitis C virus (HCV) infection among drug users and blood donors. Their HCV infection was transmitted to the general population through transfusions, folk medicine involving the breaking of the integument and tattooing. Indiscreet and widespread treatment with intravenous injection using contaminated syringes and needles at that time accelerated the transmission of HCV further. An overall result was the outbreak of HCV infection in restricted subpopulations in Japan, which inevitably involved the general population. Abrogation of paid blood donation in 1968, the exclusion of blood units contaminated with hepatitis B virus (HBV) in 1973 and that of HCV since November 1989 (by the second generation tests after February 1992) decreased the risk of posttransfusion hepatitis from >50% in the 1960s to infinitely close to zero at present. Now the incidence of HCV infection in Japan is decreased to 1.8 3.5/100,000 person-years. Mother-to-baby transmission of HBV has been prevented since 1986 by a combined passive and active immunoprophylaxis of the babies at risk with hepatitis B immune globulin and vaccine. What we see today in Japan, however, is an ever-increasing incidence of hepatocellular carcinoma (HCC) that has reached almost 40/100,000 population, with males >50 years accounting for the great majority. Of the HCC cases in Japan, approximately 16% is caused by HBV infection and approximately 80% by HCV infection. The growing incidence of HCC is expected to reach a plateau around the year 2015, and then to start to decrease. The ordeal we have gone through, with special reference to the increasing incidence of HCV-associated HCC, is expected elsewhere in the world with a current profile of age-specific HCV infection like ours a few decades back. For worse or better (probably in this order), Japan is a country far advanced as regards the HCC associated with HCV infection. Our long-term experience related in detail here is hoped to help plan strategies to contain HCV infection and cope with its long-term sequelae in many other countries worldwide. PMID- 11868792 TI - Liver transplantation as a therapeutic option for hepatocellular carcinoma. AB - Better outcomes of the patients receiving liver transplantation for viral hepatitis and hepatocellular carcinoma (HCC) are achieved by improved patient selection and perioperative treatment with antiviral agents including lamivudine, ribavirin and interferon. Patient selection is accomplished by high-quality imaging as well as exclusion of patients with large tumors, obvious extrahepatic disease or macroscopic vascular invasion. Using such criteria, a 5-year survival of 92% has been reached in the Queensland Liver Transplant Service on a small number of highly selected patients with HCC. The treatment algorithm of Makuuchi has guided us in recommending resection, estimating to what extent the liver resection can be performed safely, and timing liver transplantation when it is the only option. Adult-to-adult living-donor liver transplantation is being performed safely in many centers worldwide. The transplantation of liver from living donors to HCC patients, when standard criteria for the likelihood of good outcomes are fulfilled, will increase in Japan in the near future. PMID- 11868793 TI - Interferon for decreasing the incidence of hepatocellular carcinoma in patients with chronic hepatitis C. AB - The incidence of hepatocellular carcinoma (HCC) in Japan has kept increasing during the past few decades. Persistent infection with hepatitis C virus (HCV) is the single most frequent cause of HCC in Japan at present. Interferon is the only reliable means of eradicating HCV infection, and by inference, preventing HCC associated with it. Since interferon is not effective in all the patients with chronic hepatitis C, it needs to be used with utmost discretion. The incidence of HCC is decreased not only in the patients who respond to interferon completely, accompanied by the loss of HCV RNA from serum, but also in those who obtain normal levels of aminotransferase while HCV persists after interferon. The patients who poorly respond to interferon need to be treated by other means of suppressing necroinflammatory processes in chronic hepatitis C, which is expected to retard the development of HCC. PMID- 11868794 TI - Long-term treatment of chronic hepatitis C with glycyrrhizin [stronger neo minophagen C (SNMC)] for preventing liver cirrhosis and hepatocellular carcinoma. AB - In Japan, hepatitis C virus (HCV) is the single most frequent cause of hepatocellular carcinoma (HCC), resulting in yearly deaths of over 30,000. Although the mechanism of how HCV induces HCC is not clear, persistent HCV infection and necro-inflammatory changes in chronic hepatitis C accelerate the development of liver cirrhosis and can eventuate in HCC. Hence, means of eradicating HCV as well as suppressing inflammation in the liver, even if patients stay infected with HCV, would decrease the incidence of HCC with chronic hepatitis C. For more than 40 years, a preparation of glycyrrhizin [Stronger Neo Minophagen C (SNMC)] has been used for the treatment of 'allergic' hepatitis in Japan. In 1977, intravenous injection with SNMC was started in patients with chronic hepatitis or liver cirrhosis, most of whom have turned out to be infected with hepatitis viruses. In a multicenter double-blind study, alanine aminotransferase (ALT) levels decreased in the patients who received 40 ml/day of SNMC for 4 weeks at a rate significantly higher (p < 0.001) than controls receiving placebo. Furthermore, SNMC 100 ml/day for 8 weeks improved liver histology in 40 patients with chronic hepatitis, in correlation with improved ALT levels in serum. Liver cirrhosis occurred less frequently in 178 patients on long term SNMC than in 100 controls (28 vs. 40% at year 13, p < 0.002). Finally, HCC developed less frequently in the 84 patients on long-term SNMC than in the 109 controls (13 vs. 25% at year 15, p < 0.002). Combined, these results indicate that a long-term treatment with SNMC prevents the development of HCC in the patients with chronic hepatitis. SNMC is particularly helpful in the patients with chronic hepatitis C who fail to respond to interferon and in those who cannot be treated with it for various reasons. PMID- 11868795 TI - Pharmacokinetics of an emerging new class of anticoagulant/antithrombotic drugs. A review of small-molecule thrombin inhibitors. AB - Small-molecule direct thrombin inhibitors represent a new class of anticoagulants and are emerging as antithrombotic drugs with a range of indications. The tripeptide type or peptidomimetic compounds, including argatroban, efegatran, inogatran and napsagatran, hitherto clinically studied represent a first generation of thrombin inhibitors that are pharmacokinetically characterised by relatively rapid hepato-biliary clearance and short half-lives necessitating their administration as intravenous infusion. They are not orally bioavailable because of poor enteral absorption and presystemic hepatic extraction. Melagatran can be administered subcutaneously, and a prodrug form of melagatran, ximelagatran, is at present the only oral thrombin inhibitor available. Direct thrombin inhibitors produce predictable, stable and rapidly reversible anticoagulation measurable by common coagulation assays. Significant pharmacokinetic drug-drug interactions have not been reported. Possible pharmacodynamic interactions, in terms of prolongation of plasma clotting times, with other anticoagulant drugs must be taken into account when monitoring direct thrombin inhibitors using coagulation assays. PMID- 11868796 TI - Fast withdrawal from benzodiazepines in geriatric inpatients: a randomised double blind, placebo-controlled trial. AB - OBJECTIVE: We have previously demonstrated that temporary substitution with a low dose hypnosedative drug may lead to successful withdrawal from chronic benzodiazepine (BZD) use in the majority of patients admitted to a geriatric ward. In the present study, a withdrawal programme was evaluated in which the habitual treatment with BZDs was replaced by either 1 mg lormetazepam or placebo, defining withdrawal success rate, sleep quality and withdrawal symptoms as main outcomes. METHODS: The target population was geriatric inpatients who had been taking BZDs for at least 3 months. Subjects suffering from mental disorders were excluded. Lormetazepam or placebo were randomly assigned and given in a double blind fashion. After 1 week, the replacement therapy was discontinued. Subjective estimations of sleep quality and withdrawal symptoms were registered at predefined intervals, four times in a period of 30 days, using standard questionnaires (the Pittsburgh Sleep Quality Index and the Benzodiazepine Withdrawal Symptom Questionnaire, respectively). RESULTS: The success rate was significantly higher in the lormetazepam substitution group (80% vs 50% in the placebo group, P < 0.05). Both the subjective quality of sleep and withdrawal symptoms were significantly better in the lormetazepam substitution group. Important withdrawal effects were observed in the control group in two patients with a history of chronic alcohol abuse. CONCLUSIONS: Initial replacement therapy with a low-dose BZD is preferred over placebo, since the latter alternative is associated with worse sleep quality and a lower success rate. Placebo must only be used under medical scrutiny, given the potential for unmasking delirious symptoms, especially in patients with concomitant alcoholism. PMID- 11868797 TI - Trough:peak ratio and smoothness index in the evaluation of 24-h blood pressure control in hypertension: a comparative study between valsartan/hydrochlorothiazide combination and amlodipine. AB - OBJECTIVE: The aim of this study was to measure the time-effect profiles of a once-daily administered valsartan/hydrochlorothiazide combination and amlodipine on blood pressure using various indices derived from 24-h ambulatory blood pressure (BP) monitoring. METHODS: Of the 310 randomized outpatients with uncomplicated mild-to-moderate primary hypertension, 259 (133 on valsartan/hydrochlorothiazide, 126 on amlodipine) were eligible for analysis. After a 2-week placebo wash-out period, the patients were randomly allocated to treatment with either valsartan 80 mg once daily (o.d.) or amlodipine 5 mg o.d. for 4 weeks; in the case of an unsatisfactory blood pressure response, the treatments could be respectively changed to the fixed combination of valsartan 80 mg plus hydrochlorothiazide 12.5 mg o.d. or amlodipine 10 mg o.d. for a further 8 weeks. The trough:peak ratio (global and individualized approaches) and smoothness index (i.e., the ratio between the average of the 24-hourly BP changes after treatment and the corresponding standard deviation) were calculated from 24 h ambulatory blood pressure recordings made after the placebo period and after 4 weeks and 12 weeks of active treatment. RESULTS: Both regimens effectively lowered systolic and diastolic ambulatory pressures after 4 weeks and 12 weeks (all P<0.001) but, among the responders, the valsartan/hydrochlorothiazide combination had a greater antihypertensive effect during the night-time hours after 12 weeks (P=0.03/0.02). In the responders, the placebo-adjusted, mean trough:peak ratios were 0.76/0.74 in the valsartan/hydrochlorothiazide group (n = 111) and 0.66/0.62 in the amlodipine group (n = 101). The corresponding global trough:peak ratios were 0.61/0.57 for the valsartan/hydrochlorothiazide combination and 0.56/0.56 for amlodipine. However, the between-group differences in individual or global trough:peak ratios were not significant. The smoothness index was slightly, but insignificantly, greater for valsartan/hydrochlorothiazide than for amlodipine in the responders and the groups as a whole. CONCLUSION: Valsartan/hydrochlorothiazide and amlodipine were equally effective in reducing ambulatory BP, but the valsartan/hydrochlorothiazide combination led to more homogeneous BP control during the inter-dosing interval. Trough:peak ratio and smoothness index did not reflect this finding accurately. PMID- 11868798 TI - Spironolactone: is it a novel drug for the prevention of amphotericin B-related hypokalemia in cancer patients? AB - OBJECTIVE: Nephrotoxicity is the major adverse effect of amphotericin B (AmB), often limiting administration of full dosage. Selective distal tubular epithelial toxicity seems to be responsible for the profound potassium wasting that is a major clinical side effect of treatment with AmB. Potassium depletion also potentiates the tubular toxicity of AmB. This study was designed to assess the ability of spironolactone to reduce potassium requirements and to prevent hypokalemia in neutropenic patients on AmB treatment. METHODS: In this study 26 patients with various hematological disorders were randomized to receive either intravenous AmB alone or AmB and oral spironolactone 100 mg twice daily when developing a proven or suspected fungal infection. RESULTS: Patients receiving concomitant AmB and spironolactone had significantly higher plasma potassium levels than those receiving AmB alone (P = 0.0027). Those patients receiving AmB and spironolactone required significantly less potassium supplementation to maintain their plasma potassium within the normal range (P = 0.022). Moreover, urinary potassium losses were significantly less in patients receiving AmB and spironolactone than those receiving AmB alone (P = 0.040). CONCLUSION: This study showed that spironolactone can reduce potassium requirements and prevent hypokalemia by reducing urinary potassium loss in neutropenic patients on AmB treatment. PMID- 11868799 TI - Relative lung and total systemic bioavailability following inhalation from a metered dose inhaler compared with a metered dose inhaler attached to a large volume plastic spacer and a jet nebuliser. AB - OBJECTIVE: To compare the lung and systemic delivery of salbutamol following inhalation from a metered dose inhaler (MDI), a MDI attached to a spacer (MDI+SP) and a nebuliser (NEB) using a urinary pharmacokinetic method. METHOD: Twelve healthy subjects each provided urine samples at 0, 30 min and pooled up to 24 h after the start of 5 x 100 microg salbutamol inhaled from MDI and MDI + SP and after 2.5 mg was delivered by NEB. Following nebulisation, the amount of salbutamol trapped on an exhalation filter together with that remaining in the apparatus was determined. The amount left in the spacer and that leaving the MDI mouthpiece was also determined. Thus, for all the methods, the amount available for inhalation from each study dose was determined. RESULTS: The mean (+/- SD) 30 min urinary excretion amounts of salbutamol for MDI, MDI+SP and NEB were 12.6+/ 3.5, 27.1+/-6.0 and 16.1+/-4.6 microg, respectively. The mean ratios (90% confidence intervals) for MDI+SP compared with MDI and NEB were 230.2 (186.7, 273.8) and 183.0 (146.4, 219.7) (both P values<0.001), respectively, while that between MDI and NEB was 134 (110.4, 159.1) (P < 0.05). The mean (+/-SD) 24-h urinary excretion values for salbutamol and its metabolite were 287.0+/-46.5, 198.1+/-34.7 and 253.4+/-138.3 microg, respectively. Following inhalation a mean of 202.9+/-51.5 microg was left in the spacer. Similarly, after nebulisation 1387.7+/-88.9 microg was left in the nebuliser chamber, 26.3+/-8.0 microg in the mouthpiece and 553.8+/-68.5 microg exhaled. The mean emitted dose from the MDI was 88.4+/-6.1 microg per actuation. When normalised for the amounts available for inhalation, the mean amounts of salbutamol excreted in the urine during the first 30 min were 2.86+/-0.78, 9.15+/-1.69 and 3.06+/-0.70% following MDI, MDI + SP and NEB, respectively. CONCLUSION: Five 100-microg doses inhaled from a metered dose inhaler attached to a spacer delivered more to the lungs and less to the systemic circulation than either the same doses from a metered dose inhaler used alone or five times the dose given via a jet nebuliser. Spacers should be routinely used instead of nebulisers to manage patients unless they are short of breath. PMID- 11868800 TI - Effect of roxithromycin on the pharmacokinetics of lovastatin in volunteers. AB - OBJECTIVE: To investigate the influence of concomitant administration of roxithromycin on the plasma pharmacokinetics of lovastatin. METHODS: In an open, randomized, crossover study, 12 healthy volunteers received 80 mg lovastatin orally either alone or concomitantly with 300 mg roxithromycin after 5-day pretreatment with roxithromycin 300 mg daily. Plasma concentrations of lovastatin (lactone and acid) were determined using high-performance liquid chromatography, and the pharmacokinetic parameters were estimated. RESULTS: The mean (+/- SD) pharmacokinetic parameters of lovastatin lactone with and without roxithromycin were maximum concentration (Cmax) 8.49+/-6.80/16.3+/-9.4 ng ml(-1), time to Cmax (tmax) 1.8+/-0.4/1.7+/-0.6 h, terminal plasma half-life (t1/2) 4.3+/-2.0/3.7+/ 2.5 h, area under the plasma concentration-time curve from zero to infinity (AUC0 infinity) 53+/-60/85+/-67 ng ml(-1) h. The respective parameters of lovastatin acid were Cmax 24.6+/-13.4/17.8+/-11.0 ng ml(-1), tmax 3.7+/-1.1/4.1+/-0.7 h, t1/2 3.2+/-2.5/4.3+/-2.8 h, AUC0-infinity 149+/-123/105+/-58 ng ml(-1) h. Mean bioavailability of lovastatin lactone was lower and that of lovastatin acid was higher with concomitant treatment. However, the differences were significant only with respect to lovastatin lactone (AUC and Cmax) and Cmax of lovastatin acid. CONCLUSION: Roxithromycin does not influence the pharmacokinetics of lovastatin in such a way that dosage adjustment of lovastatin seems to be necessary during co-administration. PMID- 11868801 TI - Age and sex effects on the pharmacokinetics of linezolid. AB - OBJECTIVE: To evaluate the possible effects of age and sex on the pharmacokinetics of linezolid in healthy volunteers. METHODS: A single 600-mg dose of linezolid was administered orally to young (18-40 years) and elderly (> or =65 years) healthy males and females. Blood and urine samples were collected until 48 h after dosing and assayed for linezolid concentrations using a validated high-performance liquid chromatography method. Pharmacokinetic parameters were assessed using noncompartmental methods. Comparisons of the pharmacokinetic parameters for each age and sex group were performed using a two way analysis of variance model. Pairwise comparisons were done using least-square means analysis. RESULTS: Peak plasma drug concentrations occurred within 1.5 h after linezolid administration for males and females in both age groups. However, the maximum concentration achieved differed significantly between males and females. There was no significant difference between males and females or young and elderly for mean apparent elimination rate constant or half-life. There was no difference in mean apparent oral clearance (CLPO) between the young and elderly; however, there was a significant difference between males and females. Mean CLPO for females was approximately 37% less than mean CLPO for males when not corrected for body weight. Correcting for differences in weight reduced this difference to approximately 20%. Overall, females had a slightly lower volume of distribution than males, but this was not affected by the age of subjects. CONCLUSIONS: A dose adjustment based on age and sex is not warranted due to the wide range of linezolid concentrations that are well tolerated and the relative small difference in linezolid disposition between males and females. PMID- 11868802 TI - Isoniazid is a mechanism-based inhibitor of cytochrome P450 1A2, 2A6, 2C19 and 3A4 isoforms in human liver microsomes. AB - OBJECTIVE: In order to evaluate the inhibitory effects of isoniazid on cytochrome P450 (CYP) mediated drug metabolism, the in vitro inhibitory potency and specificity as well as the reduced nicotinamide adenine dinucleotide phosphate (NADPH)-, time- and concentration dependency of isoniazid as an inhibitor of the activity of the major human CYP isoforms were studied. METHODS: Using pooled human liver microsomes, the in vitro inhibitory effects of isoniazid on CYP1A2 (phenacetin O-deethylation), CYP2A6 (coumarin 7-hydroxylation), CYP2C9 (tolbutamide hydroxylation), CYP2CI9 (S-mephenytoin 4'-hydroxylation), CYP2D6 (dextromethorphan O-demethylation), CYP2E1 (chlorzoxazone 6-hydroxylation) and CYP3A4 (midazolam 1'-hydroxylation) activities were examined. RESULTS: After a 15 min preincubation without NADPH, isoniazid reversibly inhibited microsomal CYP2C19- and CYP3A4-mediated reactions with apparent Ki values of 36 microM and 73 microM, respectively. However, isoniazid had only weak inhibitory effects on the five other CYP-mediated reactions (Ki > 110 microM). After a 15-min preincubation with NADPH, isoniazid showed an increased inhibitory potency toward CYP1A2, CYP2A6, CYP2C19 and CYP3A4 activities (Ki = 56, 60, 10 and 36 microM, respectively). In addition, the inactivation of CYP1A2, CYP2A6, CYP2C19 and CYP3A4 by isoniazid was NADPH-, time- and concentration dependent, and was characterised by Kinact values of 0.11, 0.13, 0.09 and 0.08 min(-1), and K1 values of 285, 173, 112 and 228 microM, respectively. CONCLUSIONS: As the peak plasma concentrations of isoniazid are around 30-50 microM, isoniazid at clinically relevant concentrations reversibly inhibits CYP2C19 and CYP3A4 activities, and mechanistically inactivates CYP1A2, CYP2A6, CYP2C19 and CYP3A4 in human liver microsomes. Co-administration of isoniazid and drugs that are primarily metabolised by these CYP isoforms, particularly by CYP2C19 and CYP3A4, may result in significant drug interactions. PMID- 11868803 TI - Simultaneous estimation of cyclosporin and mycophenolic acid areas under the curve in stable renal transplant patients using a limited sampling strategy. AB - OBJECTIVES: Area under the curve (AUC)-based monitoring of cyclosporin (CsA) could help to optimise therapeutic drug monitoring in certain transplant patients in addition to trough concentration monitoring. It is the method of choice for mycophenolic acid (MPA). The objective of this study was to develop a limited sampling strategy for simultaneous estimation of CsA and MPA AUCs in long-term renal transplant patients. METHODS: Twenty kidney transplant patients treated with CsA and mycophenolate mofetil were included in a pharmacokinetic study more than 6 months after transplantation. Multilinear regression analyses were performed to develop a model enabling the estimation of both drugs' AUCs using a limited number of samples. Dose-normalised data were used throughout the analysis. RESULTS: Trough concentrations of MPA were poorly correlated with AUC, either used alone (r2 = 0.232) or together with other concentrations. Several models for CsA AUC estimation met the predefined criteria (r2>0.9, P<0.05). The AUC of MPA was best estimated by a three-concentration model (AUC=0.58 C20 min+ 0.97 C1 h + 6.64 C3 h + 3.48; r2 = 0.946). These sampling times also applied to CsA AUC (AUC = 1.17 C20 min + 0.68 C1 h + 5.36 C3 h + 4.24; r = 0.985). AUCs estimated using these models in our patients using the jack-knife procedure were found to be precise and unbiased as compared with reference trapezoidal AUCs. CONCLUSION: We were able to develop a multilinear regression model for simultaneous estimation of both CsA and MPA AUCs using only three blood samples taken up to 3 h post-dosing. PMID- 11868804 TI - Treatment patterns among newly diagnosed heart failure patients in general practice. AB - AIM: To examine treatment patterns of heart failure (HF) in general practice and to evaluate factors influencing the choice of that treatment, such as age, sex, severity of disease and co-morbidity. METHODS: We used previously identified and confirmed incident cases of HF (patients aged 40-84 years) from the General Practice Research Database in the UK (n = 938). We collected recorded information on demographics. co-morbidity and drug treatment prescribed 1 year before and after the incident diagnosis in 1996. RESULTS: Most of the study cohort was over 60 years old and presented with several concomitant diseases. Use of most cardiovascular drugs significantly increased after the diagnosis of HF. There was a greater than threefold increase in the use of angiotensin-converting enzyme inhibitors, while use of nitrates decreased after diagnosis of HF among men. Use of beta-blockers and calcium channel blockers slightly decreased after diagnosis. CONCLUSION: We found that well-established treatment practices were followed by general practitioners. Severity of the disease favoured use of diuretics and a previous ischaemic heart disease and hypertension favoured use of beta-blockers and aspirin. Women and elderly patients were less likely to receive treatment with angiotensin-converting enzyme inhibitors. PMID- 11868806 TI - Gene expression profiling of the human prostate androgen response program. PMID- 11868805 TI - Hormonal regulation of spermatogenesis in primates and man: insights for development of the male hormonal contraceptive. PMID- 11868807 TI - Policy statement on recontacting artificially inseminated patients: scientific and ethical foundations. PMID- 11868808 TI - The art of donor gamete cryobanking: current considerations. PMID- 11868809 TI - The adverse effect of sulfasalazine on spermatogenesis and male reproductive potential. PMID- 11868810 TI - Deoxyribonucleic acid replication and germ cell apoptosis during spermatogenesis in the rabbit. AB - A deeper understanding of the occurrence of some events, such as DNA replication or germ cell apoptosis during spermatogenesis, is important for improving our understanding of seminiferous epithelium biology. However, studies on these events have been restricted to small rodents. This paper analyzes the occurrence of DNA synthesis by bromo-deoxy-uridine labeling, as well as the incidence of germ cell apoptosis by the TUNEL assay in the rabbit. In contrast to previous descriptions of spermatogonium proliferation in this animal, our results indicate that DNA replicates at stages 8/1, 2, 4, 5, 6, and 7, which are equivalent to the stages at which DNA synthesis occurs in rats and mice. Apoptosis of both spermatogonia and spermatocytes generally occurred at stages 2 and 4, when late spermatocytes end the pachytene stage or undergo meiotic divisions, and these results are consistent with previous observations in the rat testis. PMID- 11868811 TI - Binding regulation of porcine spermatozoa to oviductal vesicles in vitro. AB - In vivo, en route to the site of fertilization, mammalian spermatozoa bind to oviduct epithelial cells (OECs). This binding may favor sperm survival and capacitation, but little is known about the regulation of detachment of sperm from the oviduct in vivo. Therefore, we studied sperm-oviduct interaction in vitro using vesicles formed from OECs in primary culture. Porcine oviducts were collected from gilts at the slaughterhouse. OECs were obtained by compressing the oviduct and culturing them in TCM-199 medium with 10% fetal calf serum for 48 hours. For the first experiment, to test the hypothesis that progesterone (P4) and estradiol (E2) affect sperm-OEC binding, OECs were pretreated for 48 hours with 100 ng/mL of P4 or E2. In the second experiment, porcine follicular fluid (pFF, 5%), caffeine (1 microM), calcium ionophore A23187 (1 microM), and DMSO (0.01%) were added to the incubation medium to provide insights on the mechanisms of sperm release from the oviduct. For both experiments, 50 x 10(6) sperm were coincubated with 50 microL of OEC vesicles in 1 mL of incubation medium for up to 24 hours at 37 degrees C and 5% CO2. After an initial 30 minutes of coincubation, the vesicles settled and a sample of the supernatant was removed to evaluate sperm release and acrosomal status; subsequent samples were removed after 2, 4, and 24 hours of coincubation. To evaluate the effect of the different treatments on sperm integrity, the acrosome status of the spermatozoa was determined using fluoresceinlabeled Pisum sativum agglutinin staining. Experiment 1 showed that P4 pretreatment of OECs interferes with sperm binding compared with pretreatment with E2 or controls (P < .05). In experiment 2, coincubation in the presence of A23187 increased sperm detachment compared with pretreatment with DMSO, pFF, caffeine, or controls (P < .05). For each experiment, the treatments did not affect the percentage of acrosome-reacted sperm compared with controls (P < .05). PMID- 11868812 TI - Effect of different incubation conditions on phosphatidylserine externalization and motion parameters of purified fractions of highly motile human spermatozoa. AB - The objective of this study was to assess the temporal effects of sperm incubation at body temperature with various amounts of human serum albumin (HSA) on motion parameters and phosphatidylserine externalization, an expression of membrane integrity. Purified sperm populations were prepared by discontinuous gradient separation, incubated at 37 degrees C in 3 different culture conditions (human tubal fluid [HTF] alone, HTF plus 0.3% HSA, and HTF plus 3% HSA) and evaluated at 0, 1, 3, 6, and 24 hours. Annexin V binding was used to monitor membrane translocation of phosphatidylserine and a computer-assisted semen analyzer was used to evaluate motion parameters. All incubation conditions led to a time-dependent, significant decline in sperm motion parameters and an increase in exposure of phosphatidylserine (annexin V+, live cells) to the outer leaflet of the plasma membrane in both patients and donors. Patients had a higher degree of motility loss and externalization of phosphatidylserine than donors. The decline in the percentage of normal cells (annexin V-, live) was greater in HTF alone up to 6 hours, and the decline in the percentages of motile and rapid sperm were greater in HTF alone throughout 24 hours when compared with HSA supplementation. We conclude that prolonged incubation of purified populations of highly motile human spermatozoa at body temperature was associated with significant motility loss and membrane changes as revealed by phosphatidylserine translocation. A higher concentration of HSA resulted in a relative protective effect against such impairments, particularly during the first 6 hours of incubation. Under the experimental conditions tested, significant differences were observed between infertile men and fertile controls. PMID- 11868813 TI - Enolase isoforms activities in spermatozoa from men with normospermia and abnormospermia. AB - Total enolase as well as enolase-alphaalpha. (ENO-alphaalpha, ubiquitous) and enolase-S (ENO-S, sperm-specific) activities were measured in total and Percoll selected sperm from 30 normospermic fertile men and 20 abnormospermic infertile patients. The total enolase activity was significantly higher in total sperm from patients with abnormospermia compared with normospermic patients (11.1 +/- 1.9 vs. 4.8 +/- 0.5 mlU/10(7) sperm P < .05). ENO-alphaalpha activity was significantly higher in total sperm from abnormospermic men than from normospermic men (P < .05). ENO-alphaalpha activity in Percoll-selected sperm was significantly lower compared with total sperm in both group of patients; however, for the same sperm fraction ENO-alphaalpha activity did not differ between normospermic and abnormospermic men. ENO-alphaalpha activity was related to the cell contamination ratio and to the percentage of spermatozoa with abnormal morphology. Furthermore, ENO-alphaalpha was positively correlated with the percentage of immature sperm showing an excess of residual cytoplasm. ENO-S activity was significantly higher in total sperm from normospermic patients than from abnormospermic patients (P < .05). ENO-S activity in Percoll-selected sperm was not significantly different compared with total sperm in both group of patients. However, this activity was significantly lower in Percoll-selected sperm from abnormospermic men compared with normospermic men (P < .05). ENO-S activity was not related to the cell contamination ratio but was significantly correlated with the percentage of spermatozoa with normal morphology. The 2 enolase isoforms seem to reflect 2 opposite aspects of sperm cells quality: ENO alphaalpha is associated with abnormal spermatozoa, immature spermatozoa, or both; and ENO-S is associated with normal spermatozoa. As an additional index to distinguish normal from abnormal semen, the ENO-S:ENO-alphaalpha ratio was evaluated for total and Percoll-selected sperm in both groups. This ratio seems to be a new, valuable marker of the global sperm quality in a given semen sample, and may represent a predictive index of sperm fertilizing potential. PMID- 11868815 TI - Postweaning exposure to gossypol results in epididymis-specific effects throughout puberty and adulthood in rats. AB - Gossypol, a yellow pigment found in cottonseeds, well known for its antifertility properties in animals, has been used as a contraceptive by men. The aims of this work were to evaluate the effects of gossypol throughout sexual development of male rats and to provide additional data to clarify the target site or sites of this compound in the male reproductive system. Gossypol (15 mg/kg per day) was given to animals from weaning through prepuberty (41 days), early puberty (51 days), puberty (61 days), and sexual maturity (91 days). Ventral prostate weight and fructose levels were similar in control and treated rats, suggesting that androgen levels were normal. No histological effects on the testis were detected, but there was a significant decrease in the sperm concentration in the cauda epididymidis of gossypol-treated animals killed at 61 and 91 days, as well as a significant increase in abnormal sperm in the vas deferens of treated animals. Moreover, the histology of the cauda epididymidis of the rats treated throughout puberty (ie, until days 51 and 61) showed a great number of round bodies in the lumen of the epididymis. These structures stained for the epididymis-specific protein E. Collectively, the data demonstrate that the epididymis is a target of gossypol when postweaning exposure extends throughout pubertal development, and that whereas more subtle histological effects commence around puberty, indicators reproductive competence are compromised in adulthood. PMID- 11868814 TI - Importance of glycosylation and disulfide bonds in hyaluronidase activity of macaque sperm surface PH-20. AB - PH-20 is a glycoprotein located on the surface of the sperm plasma membrane and on the inner acrosomal membrane. The best understood function of sperm surface PH 20 is its hyaluronidase activity, which results in hydrolysis of the hyaluronic acid-rich cumulus matrix during sperm penetration of this extracellular oocyte investment. In this study, we investigated whether alterations in the secondary and tertiary structures of sperm surface PH-20 would affect its enzyme activity. Proteins were isolated from the sperm plasma membrane by treatment of living cells with phosphatidylinositol-specific phospholipase C (PI-PLC). PH-20 was purified from the PI-PLC released proteins by immunoaffinity chromatography. Two dimensional electrophoresis of purified PH-20 revealed 6 isoforms with isoelectric points ranging from 5.1 to 6.0. Removal of the N-linked glycans from PH-20 with N-glycosidase F shifted the molecular weight from 64 kd to approximately 54 kd, its deduced molecular weight based on sequence analysis, suggesting that most if not all, of the potential N-glycosylation sites are linked to oligosaccharides. The lectins Con A and PSA recognized purified sperm surface PH-20 after Western blotting, suggesting that mannose is a major sugar within or at the terminal end of the linked glycan. The lectins UEA and LPA did not recognize PH-20 Western blot, suggesting that fucose and sialic acid are not terminal sugars of sperm surface PH-20. Deglycosylation of sperm surface PH-20 resulted in a complete loss of its hyaluronidase activity. The reduction of disulfide bonds with beta-mercaptoethanol or dithiothreitol also resulted in loss of enzyme activity. We conclude that the hyaluronidase activity of sperm surface PH-20 is dependent on structural features established by sulfhydryl linkages, as well as glycosylation. PMID- 11868816 TI - The testis as a major source of circulating inhibins in the male equine fetus during the second half of gestation. AB - Immunolocalization of the inhibin (a) and inhibin/activin (beta3A and betaB) subunit proteins in equine fetal testes was investigated to determine the ability of the fetal testis to produce inhibins at 120, 150, 200, and 250 days of gestation. In addition, concentrations of immunoreactive (ir)-inhibin, inhibin pro-aC, and inhibin A in both the maternal and fetal circulation were measured. It was found that plasma concentrations of ir-inhibin, inhibin pro-alphaC, and inhibin A were much higher (P < .05) in the fetal than in the maternal circulation at any stage of gestation examined. Similarly, while fetal testicular homogenate contained increased amounts of inhibins, the inhibins were undetectable in homogenates of maternal ovaries and placentae. At 120 days of gestation, all 3 subunit proteins were localized to the interstitial cells, while the immunoreactivity for the inhibin/activin 3B subunit protein was also observed in Sertoli cells. The intensity of immunoreactivity for the 3 subunit proteins in interstitial cells increased as pregnancy advanced to day 200, and, at this stage, immunoreactivity for the inhibin alpha subunit protein was observed in the fetal testes in a pattern consistent with localization in Sertoli cells. Thus, the inhibin/activin betaA subunit protein was confined to interstitial cells during the gestational periods examined. We conclude that equine fetal testes secrete large amounts of inhibins, including dimeric inhibin A and possibly other dimeric forms, such as inhibin B and activins, into the fetal circulation. These results suggest that these proteins may play some important roles in the development of fetal testes during gestation. PMID- 11868817 TI - Expression and localization of the Na+/H+ exchanger isoform NHE3 in the rat efferent ducts. AB - The efferent ducts reabsorb most of the fluid released with spermatozoa from the testis. This absorptive capacity results in a severalfold increase in sperm concentration in the proximal epididymis and is partly responsible for maintenance of the optimal microenvironment for the sperm maturation. The fluid absorption is coupled to active Na+ transport and is inhibitable by amiloride, both of which suggest a role for a Na+/H+ exchanger (NHE). NHE3 is an apical membrane NHE responsible for sodium absorption in renal proximal tubule and intestinal epithelium. In the present study, we examined the expression of NHE3 messenger RNA (mRNA) and protein in the rat efferent ducts by reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting and the localization of NHE3 by indirect immunofluoresce. RT-PCR indicated the expression of NHE3 mRNA, and Western blotting showed an NHE3 protein in the efferent duct membrane homogenate. By immunofluorescence, NHE3 was localized to the apical membrane of the nonciliated cells in the efferent duct epithelium, which also expressed aquaporin-1 water channel protein. These results suggest that NHE3 potentially plays an important role in the fluid reabsorption in the efferent ducts. PMID- 11868818 TI - Evaluation of human sperm function after repeated freezing and thawing. AB - Sperm storage via freezing has been useful for men who have difficulty masturbating during assisted reproductive technology (ART) programs and before impotency caused by chemotherapy, vasectomy, and other procedures. Studies were undertaken to evaluate the extent of cryoinjury to sperm after repeated freezing and thawing. The results showed that normozoospermic and oligozoospermic sperm survived after 3 repeated freeze-thaw cycles. The inclusion of seminal plasma did not seem to protect human sperm during freezing and thawing. There were no significant differences in recovery percentages for motile, vital, and morphologically normal sperm between slow and rapid freezing methods in thaws 1, 2, and 3 of normozoospermic and oligozoospermic unwashed (u), washed (w), and washed + seminal plasma (ws) samples. However, there were significant percentage drops in the recovery of motile and vital sperm between each thaw (ie, first to second thaw, and second to third thaw) using both slow and rapid freezing for u, w, and ws samples (P < .01). There were also no significant differences in percentage recovery of motile, vital, and morphologically normal sperm between u, w, and ws samples during thaws 1 to 3 in the normozoospermic and oligozoospermic groups. Sperm were capable of fertilizing hamster oocytes microinjected with single sperms after 3 freeze-thaw cycles as evidenced by the formation of 2 distinct pronuclei and 2 polar bodies in 22.2% and 17.2% of normozoospermic and oligozoospermic samples, respectively. The numbers of normal vital motile sperm after 3 serial freeze-thaw cycles are adequate for bringing about fertilization via intracytoplasmic sperm injection in ART programs. Thus, leftover washed sperm in laboratories that perform in vitro fertilization can be frozen, thawed, and refrozen several times without loss of the sperms' ability to fertilize. This approach has tremendous benefits for men who have difficulty producing sperm and for those with low and declining sperm counts. PMID- 11868819 TI - Differences in sperm motion between high- and low-shuttlebox avoidance rats (Hatano strains). AB - Sperm from the caudal epididymis of 2 inbred strains of Sprague-Dawley (SD) rats, selected on the basis of their high- or low-shuttlebox avoidance responses, were analyzed for motion characteristics by a computer-assisted sperm motion analysis (CASA) system. Sperm motion in high-avoidance animals (HAA) was characterized by high velocities, high amplitude of lateral head displacement (ALH), and low beat cross frequency (BCF). Conversely, sperm from low-avoidance animals (LAA) displayed low velocities, low ALH, and high BCF. These characteristics in sperm motion were not changed by washing. Furthermore, after treatment with alpha chlorohydrin (aCH) as a male antifertility agent affecting rat epididymal sperm motion, sperm velocities in HAA rats were significantly reduced to levels similar to those in untreated LAA rats. However, ALH and BCF in HAA rats treated with aCH were different from those in untreated LAA rats. Sperm adenosine triphosphate (ATP) content was higher in HAA than in LAA rats, correlating with values of their sperm velocities. These data suggest there are apparent strain differences in sperm motion between HAA and LAA rats and that these differences are dependent on factors, including sperm energy production. PMID- 11868820 TI - Detection of lipid peroxidation in equine spermatozoa based upon the lipophilic fluorescent dye C1l-BODIPY581/591. AB - The lipophilic fluorescent probe, 4,4-difluoro-5-(4-phenyl-1 ,3-butadienyl)-4 bora-3a,4a-diaza-s-indacene-3-undecanoic acid (C11-BODIPY581/591) was used to evaluate changes in lipid peroxidation in equine spermatozoa during both short term exposure to ferrous sulfate and sodium ascorbate in the presence of cumene hydroperoxide as well as during storage of spermatozoa at 5 degrees C for 48 hours. Peroxidation of C11-BODIPY581/591 was accompanied by a shift in fluorescence from red to green, and the relative amount of nonoxidized probe was determined as the ratio of red:(red + green) fluorescence as detected by either fluorescence microplate reader or by flow cytometry. The addition of Fe2SO4 (0 to 0.5 mM), low concentrations of sodium ascorbate, and the addition of cumene hydroperoxide increased peroxidation of C11-BODIPY581/591. The addition of high concentrations (10 or 20 mM) of sodium ascorbate or alpha-tocopherol reduced peroxidation of C11-BODIPY581/591 during short-term incubations. During storage at 5 degrees C in a skim milk-based extender, equine spermatozoa demonstrated a progressive decline in motility and a small but significant increase in lipid peroxidation based upon ratiometric analysis of C11-BODIPY581/591. The addition of Fe2SO4 increased lipid peroxidation in cooled spermatozoa in a dose-dependent fashion and decreased sperm motility. The addition of alpha-tocopherol, however, did not reduce lipid peroxidation during cooled semen storage. These data demonstrate that the lipophilic fluorescent probe C11-BODIPY581/591 is a useful measurement of lipid peroxidation in equine spermatozoa and that there is an increase in lipid peroxidation during cooled storage of equine spermatozoa that is increased in the presence of ferrous promoters. PMID- 11868821 TI - Aneuploidy and chromosome breakage in swim-up versus unprocessed semen from twenty healthy men. AB - Toxicological and epidemiological studies have investigated several factors that are believed to induce cytogenetic damage in human sperm cells in an effort to estimate heritable risk to future generations. Most of these studies have not differentiated damage based on cell fertility or motility. In the clinical setting, intracytoplasmic sperm injection (ICSI) bypasses the natural process of sperm selection. Although practitioners attempt to select motile sperm for ICSI, the sperm may not always demonstrate motility, maturity, or even viability. Knowing whether cytogenetic damage differs in motile versus unselected sperm would improve our ability to estimate heritable risk and lead to improved ICSI procedures, and would expand the body of toxicology and epidemiology research. We divided semen samples from 20 healthy donors and compared aneuploidy and chromosome breakage in sperm cells gathered directly from the ejaculate (unprocessed semen) with cells enriched for motility using the swim-up assay. Sperm fluorescence in situ hybridization was used to detect aneuploidy for chromosomes 13, 18, 21, X, and Y. Tandem labeling probes were used to detect breakage in the 1cen-1q12 region of chromosome 1. The occurrence of disomy 18-18 and XY18 was significantly lower in specimens enriched for motility (P = .004 and P = .001, respectively). Sperm that carried duplication errors and diploid sperm were also seen less frequently in semen analyzed by the swim-up assay (P < .008). Chromosome 1 breakage did not differ between swim up-assayed and unprocessed specimens. Findings suggest that unprocessed semen may overestimate heritable aneuploidy risk in sperm biomarker studies, and may be biologically relevant to ICSI in disomy categories 18-18 and XY18, demonstrating 1.4-fold to 1.8-fold differences. PMID- 11868822 TI - Laser oximetry: A novel noninvasive method to determine changes in penile hemodynamics in an anesthetized rabbit model. AB - This study was designed to determine the utility and validity of laser oximetry in measuring changes in penile hemodynamics. Anesthetized male New Zealand White Rabbits were divided into 2 groups, and penile hemodynamics were assessed by either laser oximetry (oxyhemoglobin, deoxyhemoglobin concentration, and oxygen saturation) or intracavernosal pressure (ICP) monitoring during penile erection induced by pelvic nerve stimulation (PNS) or intracavernosal administration of phentolamine, nitroprusside, papaverine, or sildenafil. PNS caused significant frequency-dependent increases in penile ICP. PNS also caused significant increases in penile tissue oxyhemoglobin concentrations and tissue oxygen saturation in a frequency-dependent manner. The changes in oxyhemoglobin concentrations and oxygen saturation correlated with frequency-dependent increases in ICP. Intracavernosal vasoactive drug administration produced significant increases in ICP, tissue oxyhemoglobin concentration, oxygen saturation, and duration of response as a function of increasing drug concentration. Laser oximetry permits reproducible and valid assessment of changes in penile hemodynamics comparable to conventional ICP measurements. Thus, we consider laser oximetry a reliable technique in evaluating penile hemodynamics. Its sensitivity in detecting small changes in oxyhemoglobin concentration and its noninvasive nature make it advantageous over invasive methods such as ICP monitoring and laser Doppler flowmetry. PMID- 11868823 TI - Human bronchial epithelium expresses interleukin-9 receptors and releases neutrophil chemotactic factor. AB - Growing evidence obtained from human genomic analysis and antigen-challenged transgenic mice suggests that interleukin-9 (IL-9) is a candidate factor in immunoglobulin E (IgE) production and thus is thought to be associated with bronchial inflammation and bronchial hyperresponsiveness (BHR). To evaluate the expression of the IL-9 receptor and its effect on the IL-9 human bronchial cell line BEAS-2B cells, reverse transcriptase-polymerase chain reaction (RT-PCR), immunohistochemical investigation, and chemotaxis assay were performed. The components of the IL-9 receptor, consisting of IL-9 receptor alpha (CD129) and IL 2 receptory ((1)132), were expressed on BEAS-2B cells as determined by RT-PCR and flow cytometry. BEAS-2B cells exposed to IL-9 released neutrophil chemotactic activity (NCA) in a time- and dose-dependent manner, and the presence of granulocyte colony-stimulating factor (G-CSF) was also detected. This factor is primarily involved in NCA for the measurement of cytokines and in the inhibition assay of neutrophil chemotaxis. These findings suggest that bronchial epithelial cells may express IL-9 receptors, and that IL-9 may induce airway inflammation through the release of G-CSF from bronchial epithelial cells. PMID- 11868824 TI - Effect of growth factor-fibronectin matrix interaction on rat type II cell adhesion and DNA synthesis. AB - Type II cells attach, migrate, and proliferate on a provisional fibronectin-rich matrix during alveolar wall repair after lung injury. The combination of cell substratum interactions via integrin receptors and exposure to local growth factors are likely to initiate the signals required for cell proliferation, differentiation, reepithelialization, and ultimate restoration of the alveolar wall structure. Accordingly, primary cultured type II cells have been shown to bind fibronectin, in part through the alpha5beta1 integrin, and to respond to growth factors that induce type II cell proliferation, such as fibroblast growth factor 1 (FGF-1). The purpose of this study was to determine whether or not FGF-1 modifies type II cell attachment to fibronectin, and if together they affect DNA synthesis. Attachment assays showed that FGF-1 treatment enhanced type II cell adhesion to fibronectin. This effect correlated with an increase in beta1 integrin cell surface expression, and with the formation of cytoskeletal stabilizing structures such as lamellipodial extensions and stress fibers. FGF-1 also induced an increase in thymidine incorporation into DNA. Together FGF-1 and fibronectin appear to promote adhesion, cytoskeletal organization, and increased DNA synthesis, and in this way influence cell-substratum interactions and signaling during alveolar repair. PMID- 11868825 TI - Secretory leukocyte proteinase inhibitor and elafin are resistant to degradation by MMP-8. AB - The naturally occurring neutrophil elastase inhibitors, alpha1-proteinase inhibitor (alpha1PI), secretory leukocyte proteinase inhibitor (SLPI), and elafin, are potential therapeutic agents in the treatment of neutrophil-mediated lung disease. However alpha1PI has been shown to be susceptible to inactivation by matrix metalloproteinases (MMPs) released by neutrophils, particularly neutrophil collagenase (MMP-8). The aim of this study was to determine if SLPI and elafin are similarly susceptible to degradation by this neutrophil-specific MMP. The effect of MMP-8 on SLPI and elafin was assessed by determining the neutrophil elastase inhibitory capacity (NEIC) and electrophoretic protein profile of both inhibitors following exposure to purified MMP-8. As a positive control, the effect of MMP-8 alpha1PI was assessed in parallel. Although treatment of alpha1PI with MMP-8 resulted in a significant decrease in its NEIC (P = .025), no similar decrease was observed with SLPI or elatin. Electrophoretic analysis confirmed digestion of alpha1PI by MMP-8 but no digestion of either SLPI or elafin was observed. These results demonstrate that SLPI and elafin are resistant to proteolytic inactivation by MMP-8, a property that may enhance their therapeutic application in neutrophil-mediated inflammatory lung disease. PMID- 11868826 TI - Effects of simultaneous exposure of surfactant to serum proteins and free radicals. AB - Free radicals (FRs) and serum proteins have both been implicated in the pathophysiology of surfactant dysfunction during acute lung injury (ALI). This study examines how these 2 distinct mechanisms interact to contribute to altered surfactant function in this setting. Calf lung surfactant (2 mg/mL) was incubated with no additives (C = control), and with low = (LD = 125 microM FeCl2; 250 microM H2O2) and high-dose (HD = 250 microM FeCl2, 500 microM H2O2) Fenton reaction reagents to generate hydroxyl radical. Each condition was studied with (1) no protein (N); and with 25%, 200%, and 800% (weight protein/weight phospholipid) protein added as (2) bovine albumin, (3) bovine fibrinogen, (4) hemoglobin, or (5) calf serum. Lipid (LFR) and protein (PFR) free-radical products, and modifications in the tertiary structure of Surfactant Protein A (SPA) on Western blot, were observed in N LD and N HD samples. Added proteins reduced LFR and PFR changes as well as SPA structural changes. Protection was greatest for fibrinogen, hemoglobin, and serum, and least for albumin. Minimal to no dysfunction, assayed by pulsating surfactometry, was observed in all samples. These findings indicate that addition of serum proteins to surfactant at 2 mg/mL protects against, rather than promotes, FR-mediated chemical changes in surfactant lipid and protein constituents. PMID- 11868827 TI - A matter of life and death. PMID- 11868828 TI - Modified NTP. PMID- 11868829 TI - Problem patients. PMID- 11868830 TI - Age and caries. PMID- 11868832 TI - Have you ever received training in forensic dentistry, and are you interested in receiving such training? PMID- 11868831 TI - More about licensure. PMID- 11868833 TI - Subject: Making e-mail work for you. PMID- 11868834 TI - Prevalence and trends in enamel fluorosis in the United States from the 1930s to the 1980s. AB - BACKGROUND: The National Survey of Dental Caries in U.S. School Children: 1986 1987 conducted by the National Institute of Dental Research, or NIDR, remains the only source of national data about the prevalence of enamel fluorosis. The authors analyze these data and describe changes in the prevalence of enamel fluorosis since the 1930s, as reported by H. Trendley Dean. METHODS: A sample of children comparable to those described in the 1930s was selected from the NIDR data set among children living in households served by public water systems during the child's first eight years of life. The type of water system (that is, natural, optimal and suboptimal) for each household had been recorded in the NIDR data set using data from the 1985 U.S. Fluoridation Census. The NIDR data set included information about the children's history of fluoride exposure obtained from parents. RESULTS: In the 1986-1987 period, the prevalence of enamel fluorosis (ranging from very mild to severe) was 37.8 percent among children living in residences with natural fluoride (0.7 to 4.0 parts per million fluoride ions, or F-), 25.8 percent in the optimal fluoride group (0.7 to 1.2 ppm F- and 15.5 percent in the suboptimal fluoride group (< 0.7 ppm F-). The largest increase in fluorosis prevalence from the 1930s to the 1980s was in the suboptimal fluoride group (6.5 to 15.5 percent). CONCLUSIONS AND CLINICAL IMPLICATIONS: Exposure to multiple sources of fluoride may explain the increase in enamel fluorosis from the 1930s to the 1980s. The exposure to fluoride from sources such as dietary supplements has decreased since the 1980s because of reductions in the recommended dosage, but these changes occurred too late to have an effect on the study cohort. Evidence of simultaneous use of systemic fluorides indicates the need to reinforce guidelines for the appropriate use of fluorides and promote research on measuring total fluoride exposure. PMID- 11868835 TI - Use of dental services: an analysis of visits, procedures and providers, 1996. AB - BACKGROUND: While many studies have provided data on Americans' access to dental care, few have provided a detailed understanding of what specific treatments patients receive. This article provides detailed information about the types of dental services that Americans receive and the types of providers who render them. METHODS: The authors provide national estimates for the U.S. civilian noninstitutionalized population in several socioeconomic and demographic categories regarding dental visits, procedures performed and the types of providers who performed them, using household data from the 1996 Medical Expenditure Panel Survey, or MEPS. RESULTS: Data show that while the combination of diagnostic and preventive services adds up to 65 percent of all dental procedures, the combination of periodontal and endodontic procedures represents only 3 percent. Additionally, while 81 percent of all dental visits were reported as visits to general dentists, approximately 7 percent and 5 percent of respondents who had had a dental visit reported having visited orthodontists or oral surgeons, respectively. CONCLUSION: MEPS data show the magnitude and nature of dental visits in aggregate and for each of several demographic and socioeconomic categories. This information establishes a nationally representative baseline for the U.S. population in terms of rates of utilization, number and types of procedures and variations in types of providers performing the procedures. These nationally representative estimates include data elements that describe specific dental visits, dental procedures and type of provider, and they offer details that are useful, important and not found elsewhere. PRACTICE IMPLICATIONS: By understanding these analyses, U.S. dentists will be better positioned to provide care and better meet the dental care needs of all Americans. PMID- 11868837 TI - Dens evaginatus: a diagnostic and treatment challenge. AB - BACKGROUND: Dens evaginatus, or DE, is an uncommon dental anomaly, in which an extra cusp or tubercle protrudes from the occlusal surface of posterior teeth, as well as the lingual surface of anterior teeth. Tubercles frequently are susceptible to fractures and pulpal complications. CASE DESCRIPTION: A 9-year-old Hispanic girl presented for six emergency visits over a five-week period. Her chief complaint was of pain associated with a maxillary primary second premolar. Clinically, the tooth appeared to be noncarious, but it had an abnormal-appearing root. It was extracted after the patient presented with a left facial cellulitis. CLINICAL IMPLICATIONS: DE often challenges the practitioner with its frequent and early pulpal complications. The signs and symptoms in this case resulted in a difficult diagnosis and limited treatment options. PMID- 11868836 TI - Assessing fluoride concentration uniformity and fluoride release from three varnishes. AB - BACKGROUND: The authors investigated the fluoride content uniformity of three commercial fluoride varnishes, as well as their fluoride-release behaviors. METHODS: The authors examined 20 doses from each of two tubes of Duraphat (Colgate-Palmolive Co., New York) and Duraflor (Pharmascience Inc., Montreal), and 20 doses of individually packaged 0.25-milliliter and 0.40-mL units of CavityShield (OMNII Oral Pharmaceuticals, West Palm Beach, Fla.). Part of the dose was dissolved in chloroform, followed by fluoride extraction with distilled water. The authors painted the remaining varnish from five predetermined doses from each group onto plastic substrates for examination of fluoride release. Fluoride concentrations in the solutions were measured with a fluoride-selective ion electrode. RESULTS: One-way analysis of variance showed statistically significant differences between varnish groups. The fluoride content was more uniform in Duraphat and CavityShield than it was in Duraflor. The fluoride release profiles in terms of percentage of total fluoride released over time were different among different groups of varnishes and were similar among samples from the same test group. The authors found that Duraflor released consistently more fluoride in artificial saliva than did the other two varnishes. CONCLUSIONS: Fluoride content can vary between doses dispensed from the same tube. Uniformity also varies between different varnishes and affects the retention of fluoride in the varnish. CLINICAL IMPLICATIONS: Clinicians should be aware that the nonuniform appearance of fluoride varnish as squeezed out of the tube could indicate separation of ingredients, resulting in variation of fluoride content. PMID- 11868839 TI - An evaluation of five dental Internet portals. AB - BACKGROUND: Dental Internet portals can offer dental practitioners "one-stop shopping" for many information needs. To date, no studies have described and evaluated dental portals' services and content. METHODS: The authors evaluated five dental portals from Jan. 22, 2001, to April 5, 2001, using 90 evaluation criteria in seven categories: general, services, miscellaneous, navigation and usability, site currency, site performance and responsiveness, and site integrity. Groups of three to four dental students rated each portal. The authors rated certain criteria using commercial monitoring and analysis services. RESULTS: The portals evaluated in this study provided a wide range of services such as product purchasing, online continuing education, practice management services, news, dental practice Web pages and event calendars. Portals differed in many characteristics, such as the number of services, product pricing, discussion forum activity, navigability, reaction time in response to questions and site responsiveness. CONCLUSIONS: The implementation and usefulness of each portal's services varied. No portal can fit all needs best, and many portals change rapidly owing to the volatility of the Internet industry. Dentists should be familiar with portals' services and alternatives for using them. PRACTICE IMPLICATIONS: Portals can provide useful services to dental practitioners. Practitioners, however, should evaluate portals carefully to ensure that their needs are met optimally. PMID- 11868838 TI - A systematic review of complication risks for HIV-positive patients undergoing invasive dental procedures. AB - BACKGROUND: This systematic literature review determined the strength of evidence regarding whether patients with human immunodeficiency virus, or HIV, are at higher risk of developing complications from invasive oral procedures than similar patients without HIV. TYPES OF STUDIES REVIEWED: MEDLINE and EMBASE searches of the English literature from the early 1980s through April 2000 yielded five articles meeting the inclusion and exclusion criteria: original research, concurrent treatment of HIV-positive and HIV-negative subjects, presence of complications (for example, local or systemic infection, bleeding, alveolitis, delayed healing) resulting from extractions, orthognathic surgery, periodontal therapy, endodontic therapy, placement of dental implants, prophylaxis, or scaling and root planing. RESULTS: The authors found no studies involving orthognathic surgery, periodontal therapy, dental implants, prophylaxis, or scaling and root planing, and only one study reporting few immediate endodontic therapeutic complications. Thus, the evidence is insufficient with respect to any additional risk associated with these procedures among people with HIV/AIDS. Because of the few studies, low overall complication rates and variability in results from different analytic approaches, the authors consider the evidence to be too poor to rule in or out a meaningful relationship between HIV status and complications from tooth extractions. CLINICAL IMPLICATIONS: Limited published scientific evidence is available to guide clinicians in regard to possible increased risks of invasive oral procedures associated with the HIV status of the patient. PMID- 11868842 TI - Nonsterile latex examination gloves. PMID- 11868841 TI - Clinical effect of a new liquid dentifrice containing triclosan/copolymer on existing plaque and gingivitis. AB - BACKGROUND: Dentifrices traditionally have been formulated as pastes or gels. However, liquid varieties recently have become available The authors conducted a study to determine whether a fluoridated liquid product with added triclosan and a polyvinyl methyl ether/maleic acid, or PVM/MA, copolymer delivery system is efficacious for the control of supragingival plaque and gingivitis. METHODS: The authors conducted a six-month, double-blind, parallel, stratified clinical study in accordance with American Dental Association guidelines for the clinical evaluation of antiplaque/antigingivitis products. Subjects who qualified with a suitable amount of existing plaque and gingivitis were given one of two dentifrices: an "experimental" liquid dentifrice containing 0.3 percent triclosan, 2.0 percent PVM/MA copolymer and 0.243 percent sodium fluoride, or NaF, in a silica base; or a "placebo" dentifrice containing 0.243 percent NaF in a silica base. The authors collected subjects' plaque and gingival index scores at baseline, three months and six months. RESULTS: The results of the study indicate that the use of the experimental liquid dentifrice provided a significant reduction in existing levels of plaque and gingivitis as compared with the placebo standard dentifrice. At six months, the experimental group demonstrated a 34.9 percent reduction in plaque formation and a 25.7 percent reduction in gingivitis as compared with the placebo group (P < .001). CONCLUSIONS: When compared with the placebo, the experimental dentifrice had a significantly better ability to reduce plaque and gingivitis. When compared with results reported in the literature, this new form of dentifrice achieved results comparable in efficacy to those of traditional dentifrice pastes with matching active ingredients. CLINICAL IMPLICATIONS: This study confirms that a liquid dentifrice containing a triclosan/copolymer/NaF delivery system is efficacious in its ability to reduce existing plaque and gingivitis. PMID- 11868843 TI - Conditions associated with the use of latex gloves. PMID- 11868845 TI - Peer review records subject to disclosure. PMID- 11868840 TI - The practice of your future: creating a vision. AB - BACKGROUND: In the realm of practice planning, a practice strategy is a combination of two things: a vision of a desired future and an action plan to make that vision a reality. This article presents a process called "Appreciative Inquiry," which dentists can use to create a vision of their future practices. TECHNIQUES: In creating a vision of his or her future practice through Appreciative Inquiry, the dentist builds on the best of what currently is (in other words, the strengths of the practice), visualizes the various possibilities of what the practice could become and, finally, designs the specific elements of what the practice actually will be at its ideal in the future. CONCLUSIONS: Appreciative Inquiry offers a dynamic perspective in helping dental teams create a vibrant new vision for their ideal practices of the future--the first step in building an effective strategy. PRACTICE IMPLICATIONS: A clear vision helps a practice set its desired course for the future, helps the practice to change in ways the dental team desires and provides a framework for responding to changes in the world in which the practice exists. PMID- 11868844 TI - Preventing postoperative tooth sensitivity in class I, II and V restorations. AB - Postoperative tooth sensitivity in Class I, II and V resin-based composite restorations continues to be an unpredictable problem in dentistry. In spite of meticulous use of dentin bonding agents, dentists and patients are faced with the sensitivity problem and the frustrating need to remove restorations and occasionally accomplish endodontic therapy on teeth that were not sensitive before the restorations were placed. Practitioners have developed numerous preventive methods to overcome the sensitivity challenge, which I have described in this article. PMID- 11868846 TI - For the dental patient: baby's first teeth. PMID- 11868847 TI - Oxygen permeability (Dk) of thirty-seven rigid contact lens materials. AB - PURPOSE: Oxygen permeability (Dk) was determined for 37 available rigid contact lens materials in a masked fashion. The results were compared with those of an earlier study that included different lots of 14 test materials assessed in the current study. METHODS: Six lenses of different thicknesses in each test and reference material were obtained. Test materials were arranged in sets of six to eight materials per set. Each set of materials, with inclusion of at least two reference materials for the purpose of simultaneous calibration, was measured to obtain preliminary amperages. Four preliminary measures were performed per thickness, resulting in 24 per material, in a schedule designed to spread the potential effects of machine drift and other factors. The mean preliminary amperages were used to derive corrected Dk values according to American National Standards Institute (ANSI) Z80.20-1998, and the values were linearly calibrated using the measured and established Dk values of the reference materials. RESULTS: The resistance (t/Dk) vs. thickness (t) plots for the 37 test and seven reference materials were approximated linearly. In 54 of 57 linear regressions, the coefficients of determination (R2) were >0.96, and in 48 instances were >0.98. Fourteen Dk values from the current study and an earlier study were linearly correlated (R2 = 0.9846), with a slope close to unity (+1.056) and intercept close to zero (-0.292). Ten of the current values fell within 10% of their corresponding earlier values. Only three current Dk values fell outside of the ANSI Z80.20-1998 tolerance for Dk (+/-20%). Two of these Dk values met the product tolerance when an obvious outlying point was graphically identified and omitted from the linear resistance (t/Dk) vs. thickness (t) regression. CONCLUSION: Omission of a single outlying point from a linear resistance vs. thickness regression can help provide a more valid Dk value. The ANSI Z80.20-1998 tolerance of +/-20% on Dk and the measurement reproducibility of +/-10% were achieved for the overwhelming majority of rigid contact lens materials up to at least 160 Dk units. The corrected, calibrated Dk values for the 37 test materials, in ANSI units, ranged from 13.8 to 175.1, having an overall mean of 43.2, median of 31.9, and standard error of 6.57 (N = 37). PMID- 11868848 TI - Do specially designed visual display unit lenses create increased postural load compared with single-vision lenses during visual display unit work? AB - Three different types of spectacle lenses, specially designed for visual display unit (VDU) work, were compared with single-vision lenses regarding postural load. The different corrections effect on postural load was measured by using electromyography. Muscle loads were recorded from the trapezius muscle and the infraspinatus muscle. Continuous measurements of body posture were measured by using three dual axis inclinometers attached to the head, back, and upper arm. No significant differences were found between the single-vision lenses and the specially designed VDU lenses regarding muscle load. Small differences were found regarding the head angle. The study concludes that these new lens designs create interesting opportunities for the optometrist to optimize the visual conditions for VDU workers. PMID- 11868849 TI - Calibration error on the measurement of back vertex power for contact lenses with method using focimeter with manual focusing. AB - PURPOSE: The International Standard ISO 9337-1, which sets forth the method for measuring back vertex power of contact lenses with manually focusing focimeters, specifies that test lenses conforming to ISO 9342 may be used to calibrate focimeters on its spectacle lens support, and correction values obtained in this way can be used directly in measuring contact lenses on its contact lens support. This study was conducted because of concern that the method mentioned in ISO 9337 1 is not good enough for the calibration of focimeters used to measure contact lenses. METHODS: To test the validity of this method, a research group from China National Institute of Metrology (NIM) studied it theoretically and carried out a series of comparison experiments, respectively, with the conventional test lenses conforming to ISO 9342 and test lenses made at NIM (with an expanded uncertainty of 0.025 D). RESULTS: The results show that the measurement error between the two calibration methods will exceed 0.50 D if the specification described in ISO 9337 1 is adopted. This error also exceeds allowable tolerances for focimeters and for contact lenses themselves. Experiments and theoretical calculations done by the NIM group show that these errors mainly come from spherical aberration. Vertex error induced by the lens support is not negligible. CONCLUSION: When focimeters are calibrated to find correction values with the method specified in ISO 9337-1, measurement error will not be eliminated if these correction values are used in measuring contact lenses, and the resulting deviation is too large to be ignored. Therefore, special test lenses should be used to calibrate focimeters to find correction values when these focimeters are used to measure back vertex power of contact lenses because contact lenses are a special product being used to correct human vision. PMID- 11868850 TI - Objective criteria for the determination of exit competencies. PMID- 11868851 TI - Does mobility performance of visually impaired adults improve immediately after orientation and mobility training? PMID- 11868852 TI - Effect of dichoptic adaptation on frequency-doubling perimetry. AB - BACKGROUND: In frequency-doubling technology (FDT) perimetry, the second eye tested has reduced sensitivity. We investigated the cause of this sensitivity reduction. METHODS: Contrast sensitivity was measured for frequency doubling (0.25 cpd, 25 Hz) and nonflickering (4 cpd) gratings, arranged similarly to the targets in the FDT perimeter (Welch Allyn, Skaneateles Falls, NY, and Humphrey Instruments, San Leandro, CA). Various test orders were examined. RESULTS: Frequency-doubling (FD) sensitivity was reduced (0.15 log) in the second eye. A similar reduction occurred when first and second eye gratings were oriented orthogonally, suggesting that dichoptic contrast adaptation was not the cause. FD sensitivity was little affected after testing with nonflickering gratings, suggesting that fatigue effects were small. Sensitivity was reduced when testing was performed after 5 min of opaque occlusion. The use of a translucent occluder eliminated the reduction in sensitivity in the second eye. CONCLUSION: We confirm the presence of reduced sensitivity in the second eye tested with FD perimetry and find that it is caused by delayed light adaptation post-occlusion. PMID- 11868853 TI - Time course of masking in spatial resolution tasks. AB - PURPOSE: Further evidence is presented that Landolt C acuity has an integration time extending over several hundred milliseconds, well beyond the Bunsen-Roscoe Bloch critical duration within which the intensity-time reciprocity for visual thresholds holds. METHODS: To test whether all segments of a longer exposure contribute equally to the uptake of acuity information, the acuity impairment caused by short-duration masking stimuli was measured for a range of onset asynchronies. RESULTS: A 50-ms mask impairs resolution most when it is presented 50 to 100 ms after the onset of the target, and it no longer influences acuity when shown 50 ms after its extinction. CONCLUSION: Allowing for the well established optimally-effective test-mask asynchrony of 50 ms or so, the results permit the conclusion that whereas acuity information is assimilated during the whole exposure of a target, the first 50 ms are more effective, and this fits with the initial peaking of impulse activity of neurons in the primate visual cortex to flashed visual stimuli. PMID- 11868854 TI - Fenugreek and insulin resistance. PMID- 11868855 TI - Effect of Trigonella foenum-graecum (fenugreek) seeds on glycaemic control and insulin resistance in type 2 diabetes mellitus: a double blind placebo controlled study. AB - OBJECTIVES: To evaluate the effects of Trigonella foenum-graecum (fenugreek) seeds on glycemic control and insulin resistance, determined by HOMA model, in mild to moderate type 2 diabetes mellitus we performed a double blind placebo controlled study. METHODS: Twenty five newly diagnosed patients with type 2 diabetes (fasting glucose < 200 mg/dl) were randomly divided into two groups. Group I (n=12) received 1 gm/day hydroalcoholic extract of fenugreek seeds and Group II (n=13) received usual care (dietary control, exercise) and placebo capsules for two months. RESULTS: At baseline both the groups were similar in anthropometric and clinical variables. Oral glucose tolerance test, lipid levels, fasting C-peptide, glycosylated haemoglobin, and HOMA-model insulin resistance were also similar at baseline. In group 1 as compared to group 2 at the end of two months, fasting blood glucose (148.3 +/- 44.1 to 119.9 +/- 25 vs. 137.5 +/- 41.1 to 113.0 +/- 36.0) and two hour postglucose blood glucose (210.6 +/- 79.0 to 181.1 +/- 69 vs. 219.9 +/- 41.0 to 241.6 +/- 43) were not different. But area under curve (AUC) of blood glucose (2375 +/- 574 vs 27597 +/- 274) as well as insulin (2492 +/- 2536 vs. 5631 +/- 2428) was significantly lower (p < 0.001). HOMA model derived insulin resistance showed a decrease in percent beta-cell secretion in group 1 as compared to group 2 (86.3 +/- 32 vs. 70.1 +/- 52) and increase in percent insulin sensitivity (112.9 +/- 67 vs 92.2 +/- 57) (p < 0.05). Serum triglycerides decreased and HDL cholesterol increased significantly in group 1 as compared to group 2 (p < 0.05). CONCLUSIONS: Adjunct use of fenugreek seeds improves glycemic control and decreases insulin resistance in mild type-2 diabetic patients. There is also a favourable effect on hypertriglyceridemia. PMID- 11868856 TI - A study of plasma alpha-2-macroglobulin levels in type 2 diabetic subjects with microalbuminuria. AB - BACKGROUND: Alpha-2 macroglobulin (Alpha-2-M) is a major plasma protease inhibitor that also regulates the activity of a variety of bioactive peptides including interleukins and exerts a range of immunomodulatory effects. OBJECTIVE: We conducted the present study with the objective to study the alpha-2-M levels in type 2 diabetic subjects with microalbuminuria in an attempt to establish alpha-2-M as a predictor of microvascular complications in diabetes. MATERIAL AND METHODS: Plasma Alpha-2-M levels were assayed in 100 (53 males and 47 females) randomly selected type 2 diabetic subjects with microalbuminuria. Diabetes was diagnosed according to the expert committee report of 1998. Patients with any acute metabolic complication like hypoglycemia, ketoacidosis, cerebrovascular accident or any acute infection were not included in the study group. RESULTS: Majority of patients belonged to 40-60 years age group. In our study alpha-2-M levels indicated a clear increase in diabetic subjects with the increasing age of subjects confirmed by multiple logistical analysis. Alpha-2-M levels were not found to be significantly different between males and females (55.6 +/- 11.3 vs. 53.7 +/- 10.5). Duration of diabetes was found to be an important confounding variable showing a direct positive correlation with alpha-2-M levels and also a significant correlation was found between alpha-2-M levels with different levels of microalbuminuria on multiple logistical analysis. No significant relation of alpha-2-M levels with either fasting blood sugar or HbA1 was observed. CONCLUSION: The increase in plasma alpha-2 macroglobulin levels in diabetes may be a correlative measure to encounter the potential proteolytic challenge associated with diabetic microangiopathy, even very early in the course of the disease. Alph-2 macroglobulin may yet be one of the most specific markers of microvascular complications in diabetes than any other serum protein. PMID- 11868857 TI - Circadian pattern of cardiovascular and cerebrovascular diseases in geriatric population. AB - OBJECTIVES: Over last 13 years various studies have been done to evaluate the circadian pattern in cardiovascular and cerebrovascular diseases in adults and the existence of such variation in Indian population also has been demonstrated. The data on this variation in geriatric patients does not exist. METHODS: We undertook this prospective observational study at Government Medical College and Hospital, Chandigarh to evaluate the circadian variation in disorders like acute myocardial infarction (AMI), unstable angina (USA), non Q wave MI (non QMI), cerebrovascular accidents (strokes, both ischemic and hemorrhagic) and transient ischemic attacks (TIA). OBSERVATIONS: We studied 158 patients (56.98% males and 43.02% females), mean age was 69 +/- 4 years. 34.17% each had AMI and CVA, 22.78% and USA and 7.59% had NON Q MI, only two patients in our study group had TIAs. We divided 24 hours into four equal quarters each for analysis. RESULTS: We observed that maximum episodes were seen during the period between 6 am till 12 noon 58/158 (36.71%) and a second peak was seen during 6 pm and 12 midnight when 40/158 events were recorded (25.31%). The least number of episodes were seen during the period between 12 midnight till 6 am 22/158 (13.92%). Similar peaking of events was noted for acute myocardial infarction but only one peak was seen for unstable angina. For cerebrovascular accidents two similar peaks were noted between 6 am till 12 noon and 12 noon till 6 pm. CONCLUSIONS: Our study population (geriatric patients) shows the presence of a definitive circadian variation with two comparable peaks. One during the morning hours (6 am-12 noon) and another peak between 6 pm and 12 midnight in patients having acute coronary diseases. In cerebrovascular accidents patients too, similar peaks were noted between 6 am till 12 noon and 12 noon till 6 pm. PMID- 11868858 TI - Electrophysiological evaluation of 140 hands with carpal tunnel syndrome. AB - AIM: To study various electrophysiological parameters in clinically suspected carpal tunnel syndrome (CTS). MATERIAL AND METHODS: Electrophysiological (EPS) evaluation of 70 patients (140 hands) of clinically suspected CTS was done as per AAEM recommendation. EPS grading was done as described by Padua, et al. RESULTS: The mean age was 44.21 +/- 10.29 (range 24-66) years with female predominance (M:F 3:11). Sixty four (91.4%) patients had bilateral involvement. The mean EPS grade was 3.18 +/- 1.41. One hundred and six (75.5%) hands had moderate to severe (EPS grade 1-3) CTS. Median distal motor latency (DML) (mean 4.76 +/- 1.4 msec) was prolonged in 92 (65.71%) hands and was not recordable in seven (5%) hands. Median sensory distal latency (SDL) (mean 3.54 +/- 0.82 msec) was prolonged in 38 (27.1%) hands and not recordable in 49 (35%) hands. Sensory median mid-palm latency (MPL) (2.42 +/- 0.56 msec) was commonest EPS abnormality present in 128 (91.4%) hands. EMG was abnormal in 75% hands. CONCLUSION: This study suggests incorporation of median sensory midpalm latency as a screening test for electrophysiological diagnosis of CTS. In addition, bilateral studies are recommended as there is bilateral involvement in 64 patients. PMID- 11868859 TI - Ankle brachial index as a predictor of generalized atherosclerosis. AB - OBJECTIVE: To assess ankle brachial index (ABI) as a screening method to target subclinical atherosclerosis in middle aged individuals. MATERIAL AND METHODS: Total 160 patients over the age of 40 years were included in the study for a period of 16 months. Their ABI was determined either by colour Doppler method (30 patients) and/or sphygmomanometry (all 160 patients). A value of < 0.9 was taken as cutoff point for significant stenosis. RESULTS: Total 69 patients out of total of 160 had significantly low ABI value (43.12%) which shows that there is a very high incidence of low ABI in the community. Overall > 50% of the patients were largely asymptomatic and had presence of two or more risk factors. ABI < 0.9 was a good screening test to detect such individuals at an earlier stage (sub clinical). CONCLUSION: A significantly low (< 0.9) ABI value can detect subclinical atherosclerotic vascular involvement and predict future occurrence of preventable major vascular event. PMID- 11868860 TI - Pancytopenia--a six year study. AB - All cases of pancytopenia seen over a six year period in two hematology centres were analysed. Patients receiving myelotoxic chemotherapy or those with leukemic cells in peripheral smear were excluded. There were a total of 166 cases. The four major causes were: aplastic anaemia in 49, megaloblastic anaemia in 37, aleukemic leukemia or lymphoma in 30 and hypersplenism in 19 cases. Bone marrow aspiration was often unsuccessful in obtaining an adequate sample, while biopsy was usually diagnostic. Both bone marrow aspiration and biopsy should be performed simultaneously in pancytopenic patients when the diagnosis is elusive. Megaloblastic anaemia is a major cause of pancytopenia, and may present acutely in the critically ill. It is a rapidly correctable disorder and should not be missed. PMID- 11868861 TI - Comparison of aminophylline and insulin-dextrose infusions in acute therapy of hyperkalemia in end-stage renal disease patients. AB - OBJECTIVE: This study was performed to compare the efficacy of aminophylline and insulin-dextrose infusion as acute treatment modality of hyperkalemia in patients with end-stage renal disease (ESRD). METHODS: This study was conducted on 30 ESRD patients with serum potassium > 6.0 mEq/L. These patients were divided in two groups of 15 each. Group A patients were treated with aminophylline infusion, whereas group B patients were treated with insulin-dextrose infusion. Serum potassium and other biochemical parameters such as blood sugar were measured at beginning of treatment followed by at 60 minutes, 180 minutes, and 360 minutes after treatment. RESULTS: Intervenous infusion of aminophylline lowered plasma potassium from 6.48 +/- 0.39 mEq/L to 5.92 +/- 0.40 mEq/L at 180 minutes (p < 0.001 Vs basal) and 6.05 +/- 0.53 mEq/L at 360 minutes (p < 0.01 Vs basal). Whereas, intravenous infusion of insulin-dextrose decreased plasma potassium from 6.59 +/- 0.31 mEq/L to 5.76 +/- 0.32 mEq/L (p < 0.001 Vs basal) and 5.84 +/- 0.21 mEq/L (p < 0.001 Vs basal). Thus in both groups, plasma potassium levels were significantly less than basal levels throughout the study. The decrease in plasma potassium was significantly more in group B patients (p value is < 0.001 after 60 minutes, < 0.05 after 180 minutes and < 0.05 after 360 minutes) when compared to group A patients. There was one episode of hypoglycemia (blood sugar < 60 mg%) in insulin-dextrose infusion group. No other side effects were observed throughout the study. CONCLUSION: Aminophylline is an effective modality for acute treatment of hyperkalemia, though it is less effective than insulin-dextrose infusion. However, more studies are required to confirm these results. PMID- 11868862 TI - Safety and efficacy of liposomal amphotericin B in patients with cryptococcal meningitis. AB - Four patients with cryptococcal meningitis were successfully treated with liposomal amphotericin B prepared at our institute using Soya phosphatidylcholine and cholesterol. In one patient, response with 1 mg/kg/day treatment was poor. However, on increasing the dose to 2 mg/kg/day, a good response was observed with CSF becoming negative for Cryptococcus neoformans after seven days of this enhanced dose. L-AMP-LRC-1 was found to be well tolerated and a major advantage was observed in two renal transplant patients in whom it could be given safely. PMID- 11868863 TI - Postage stamp of France released in 1970 on 150th anniversary of discovery of quinine. PMID- 11868864 TI - Pictorial CME. A 31 year male presented with acute onset of breathlessness. PMID- 11868865 TI - Present status of various inotropic agents in heart failure. PMID- 11868867 TI - A current perspective on clubbing and hypertrophic osteoarthropathy. PMID- 11868866 TI - Lipoprotein (a): role in diabetes and its vascular complications. PMID- 11868868 TI - Chest trauma causing acute myocardial infarction. AB - We report a case in which blunt chest injury during football match caused an intimal tear in the left anterior descending coronary artery resulting into acute anterior wall myocardial infarction. PMID- 11868869 TI - Duodenal leiomyoma--a rare cause of haematemesis. AB - Leiomyoma of the duodenum is a rare tumour. Small intestinal tumours contributing to upper gastrointestinal bleed is still rare. They usually present with malena and anaemia, rarely hematemesis. We report a case of leiomyoma of duodenum diagnosed on endoscopic ultrasound that presented with massive haematemesis. PMID- 11868870 TI - Multiple myeloma presenting as proptosis and sixth nerve palsy. AB - Cranial and intracranial locations are rare in multiple myeloma (MM). But their occurrence has a particular significance. Proptosis and 6th nerve palsy is very uncommon presentation. We report a case of MM with presenting features as proptosis and 6th nerve palsy. PMID- 11868871 TI - Cholelithiasis in a child--an unusual presentation of Wilson's disease. AB - A nine year old mentally retarded girl with moderate splenomegaly and ascites presented with chronic cholelithiasis. The presence of Kayser-Fleischer rings and low serum ceruloplasmin level confirmed the diagnosis of Wilson's disease. Cirrhosis of liver and recurrent episodes of hemolysis--these two common complications of Wilson's disease make an ideal setting for gall stone formation. Only three such cases have been reported worldwide and ours is the first case report from India. We suggest that cholelithiasis and splenomegaly in a child without evidence of congenital hemolytic disease should be taken as a suspect of Wilson's disease. PMID- 11868872 TI - Systemic hypersensitivity reaction to mesacol. PMID- 11868874 TI - Streptococcus agalactiae endocarditis. AB - Streptococcus agalactiae (S. agalactiae) is a rare cause of infective endocarditis, which is associated with a high mortality rate. Endocarditis in adults is generally related to immunocompromised states. We hereby report the case of a 35 year old man who presented with fever and delirium in whom aortic valve endocarditis due to S. agalactiae was detected. Though most patients with S. agalactiae endocarditis need surgical intervention along with antibiotics, our patient improved with medical therapy alone. PMID- 11868873 TI - Non-Hodgkin's lymphoma with pure red cell aplasia. AB - Pure red cell aplasia (PRCA) associated with non-Hodgkin's lymphoma, is an extremely rare condition, with few cases reported worldwide. More commonly PRCA is associated with thymoma or viral infections. Steroids and other immunosuppressive drugs are the preferred treatment of PRCA. PMID- 11868875 TI - Kala azar in a case of chronic hepatitis B with cirrhosis of liver. PMID- 11868876 TI - Piracetam in post-hypoxic action myoclonus. PMID- 11868877 TI - Exercise induced bronchospasm in house dust mite sensitive children. PMID- 11868878 TI - Hypertension in elderly. PMID- 11868879 TI - Profile of hypertension in elderly subjects. PMID- 11868880 TI - Cranial irradiation--an unusual cause for diabetes insipidus. PMID- 11868881 TI - Lichtenberg figures. PMID- 11868882 TI - Fibrocalculous pancreatic diabetes and macrovascular disease. PMID- 11868883 TI - Juvenile Parkinson's disease. PMID- 11868884 TI - Human leptin deficiency and resistance. PMID- 11868885 TI - Change, constancy, and help wanted (from the whole village). PMID- 11868886 TI - Controlling methicillin-resistant Staphylococcus aureus: a feedback approach using annotated statistical process control charts. AB - OBJECTIVES: To investigate the benefit of a hospitalwide feedback program regarding methicillin-resistant Staphylococcus aureus (MRSA), using annotated statistical process control charts. DESIGN: Retrospective and prospective analysis of MRSA rates using statistical process control charts. PARTICIPANTS: Twenty-four medical, medical specialty, surgical, intensive care, and cardiothoracic care wards and units at four Glasgow Royal Infirmary hospitals. METHODS: Annotated control charts were applied to prospective and historical monthly data on MRSA cases from each ward and unit during a 46-month period from January 1997 through September 2000. Results were fed back from December 1999 and then on a regular monthly basis to medical staff, ward managers, senior managers, and hotel services. RESULTS: Monthly reductions in the MRSA acquisition rate started 2 months after the introduction of the feedback program and have continued to the present time. The overall MRSA rate currently is approximately 50% lower than when the program began and has become more consistent and less variable within departments throughout Glasgow Royal Infirmary. The control charts have helped to detect rate changes and manage resources more effectively. Medical and nursing staff and managers also report that they find this the most positive form of MRSA feedback they have received. CONCLUSIONS: Feedback programs that provide current information to front-line staff and incorporate annotated control charts can be effective in reducing the rate of MRSA. PMID- 11868887 TI - Molecular typing and antimicrobial susceptibility of vancomycin-resistant Enterococcus faecium in Brazil. AB - OBJECTIVES: To characterize vancomycin-resistant enterococci (VRE) isolates and to evaluate the mode of dissemination of this pathogen in Brazil. DESIGN: We collected 22 vancomycin-resistant Enterococcus faecium isolates from 6 medical centers in Sao Paulo, Brazil, and 1 isolate from a medical center in Curitiba, Brazil. PARTICIPANTS: All Brazilian hospitals that had identified vancomycin resistant E. faecium up to the beginning of this study (late 1999) contributed isolates to the study. METHODS: The isolates were susceptibility tested using the broth microdilution method and the E-test. The presence of vancomycin resistance genes (vanA, vanB, vanC1, vanC2-3, and vanD) was evaluated by polymerase chain reaction; molecular typing was performed by pulsed-field gel electrophoresis (PFGE). RESULTS: The vanA gene was demonstrated in all vancomycin-resistant E. faecium, except for 1 isolate. None of the vancomycin resistance genes cited above was detected in the isolate from Curitiba, which was the first vancomycin resistant E. faecium described in Brazil. All isolates were resistant to ampicillin and teicoplanin. The main clone remains susceptible to doxycycline and chloramphenicol, but intermediate to quinupristin-dalfopristin. PFGE analysis demonstrated 7 major PFGE patterns. A unique PFGE pattern with 4 subtypes was detected in 17 isolates from 4 different hospitals. CONCLUSION: The results of our study indicate the occurrence of intra- and interhospital dissemination of VRE in Sao Paulo, Brazil. PMID- 11868888 TI - Stool colonization with vancomycin-resistant enterococci in healthcare workers and their households. AB - OBJECTIVE: To determine the prevalence of stool colonization with vancomycin resistant enterococci (VRE) among healthcare workers (HCWs) and their families. DESIGN: Prospective assessment of fecal colonization with VRE. SETTING: A 603 bed, tertiary-care teaching hospital. PARTICIPANTS: Healthy volunteers recruited from hospital employees and their households were screened to exclude pregnancy, diabetes mellitus, immunosuppressive disorders, and recent use of antimicrobials. INTERVENTION: Self-obtained stool swabs were used to obtain cultures. Isolated enterococci were screened for vancomycin resistance and species were identified. Intra-household isolates were genotyped using pulsed-field gel electrophoresis (PFGE). RESULTS: The participants (n = 228; age range, 28 days to 80 years) were from 137 households with and 91 without employees who had contact with patients. Enterococcus species were isolated from 127 stool specimens (55.7%). VRE were detected in 12 individuals, representing 6 E. casseliflavus, 5 E. faecium, and 1 E. gallinarum. VRE were more commonly isolated in employees who had contact with patients (5 of 52 vs 0 of 40; relative risk [RR], 1.9; 95% confidence interval [CI95], 1.5 to 2.2; P = .07) and their household members (10 of 137 vs 2 of 91; RR, 3.3; CI95, 0.7 to 14.8; P = .13). In 2 households (2 adults in a physician's household and an adult plus a child in a nurse's household) PFGE analysis demonstrated identical intra-household strains of vancomycin-resistant E. faecium. CONCLUSIONS: VRE colonization was found in 5.3% of screened stools and was more prevalent in HCWs who had contact with patients and their households. Identical PFGE patterns between 2 employees who had contact with patients and their household members demonstrated probable intra-household spread. Although the mode of acquisition was uncertain, the association with employees who had contact with patients suggests possible occupational sources. These findings demonstrate the spread of VRE within the household and implicate occupational risk for its acquisition. PMID- 11868889 TI - The direct costs of nosocomial catheter-associated urinary tract infection in the era of managed care. AB - OBJECTIVE: To determine the additional direct costs of hospitalization attributable to catheter-associated urinary tract infection (CAUTI) in 1,497 newly catheterized patients. DESIGN: Prospective observational and laboratory study. SETTING: University hospital. METHODS: Data were collected on risk factors for CAUTI (defined as > 10(3) colony-forming units [CFU]/mL), severity of illness, and diagnostic and therapeutic interventions in consenting newly catheterized patients. Daily urine cultures were obtained from each newly catheterized patient, but the results of these cultures were not revealed to his or her physician. During the study, one of the investigators (DGM) reviewed each patient's record and made a judgment as to which of the diagnostic tests and treatments ordered and what incremental length of stay could reasonably be ascribed to his or her CAUTI. The total hospital costs for each patient were also obtained. RESULTS: Overall, 235 patients acquired CAUTIs during the study; most of the CAUTIs were completely asymptomatic, and only 52% were diagnosed by the patients' physicians using the hospital laboratory. Only 1 patient with a CAUTI had a secondary bloodstream infection. Thirty-three (13%) of the CAUTIs were caused by Escherichia coli; 63 (25%) by Klebsiella, Enterobacter, Citrobacter, Pseudomonas aeruginosa, or other antibiotic-resistant, gram-negative bacilli; 87 (35%) by enterococci or staphylococci; and 67 (27%) by Candida species. The 123 CAUTIs diagnosed by the hospital laboratory were judged to have been responsible for an additional $20,662 in extra costs of diagnostic tests and $35,872 in extra medication costs, a mean of $589 (median, $356) per CAUTI. CAUTIs caused by E. coli cost considerably less than infections caused by other gram-negative bacilli ($363.3 +/- $228.2 vs $690.4 +/- $783.7; P = .02) or yeasts ($821.2 +/- $2,169.9). There were less striking differences in the costs per CAUTI caused by staphylococci or enterococci ($387.1 +/- $434.8). CONCLUSIONS: The extra direct costs associated with nosocomial CAUTI found in this prospective study, which was done in the era of managed care during the late 1990s, are substantially lower than those reported in the largest comparable studies done more than 15 years ago, most of which were retrospective, reflecting the powerful impact of cost containment measures that are now implemented in managed care. PMID- 11868890 TI - Hand washing and physicians: how to get them together. AB - OBJECTIVE: To determine the motivating and behavioral factors responsible for improving compliance with hand washing among physicians. DESIGN: Five unobtrusive, observational studies recording hand washing after direct patient contact, with study results reported to physicians. SETTING: A 450-bed hospital in a health maintenance organization with an 18-bed medical-surgical intensive care unit (ICU) and a 12-bed cardiac care unit. METHODS: An infectious disease physician met individually with participants to report study results and obtain a commitment to hand washing guidelines. Follow-up interviews were conducted to evaluate behavioral factors and educational programs. Hand washing study results were presented to all staff physicians by live and videotaped inservice presentations and electronic mail (e-mail) newsletters. The importance of influencing factors and the educational effectiveness of the hand washing program were evaluated. RESULTS: Five observational hand washing studies were conducted in the ICU between April 1999 and September 2000. Rates of physician compliance with hand washing were 19%, 85%, 76%, 74%, and 68%, respectively. There were 71 initial encounters and 55 follow-up interviews with the same physicians. Physician interviews revealed that 73% remembered the initial encounter, 70% remembered the hand washing inservice presentations, and 18% remembered the e mail newsletters. Personal commitment and meeting with an infectious disease physician had the most influence on hand washing behavior. Direct inservice presentations (either live or videotaped) had more influence than did e-mail information. Rates of ventilator-associated pneumonia did not significantly change before and during the study periods. A decrease in the rate of central line-related bloodstream infections from 3.2 to 1.4 per 1,000 central-line days was found, but could not be solely attributed to improved physician compliance with hand washing. CONCLUSIONS: Physician compliance with hand washing can improve. Personal encounters, direct meetings with an infectious disease physician, and videotaped presentations had the greatest impact on physician compliance with hand washing at our medical center, compared with newsletters sent via e-mail. Local data on compliance with hand washing and physician involvement are factors to be considered for physician hand washing compliance programs in other medical centers. PMID- 11868891 TI - Sixteen years' surveillance of surgical sites in an Irish acute-care hospital. AB - OBJECTIVE: To report a program of continuous surveillance of surgical-site infections (SSIs) using basic surveillance methods. DESIGN: Analysis of routine prospective surveillance data. SETTING: Two hospitals in Ireland (300 and 350 beds) that merged and moved to a new 650-bed hospital in 1987. PATIENTS: 59,335 surgical sites of postoperative patients. INTERVENTIONS: Surgical sites were surveyed by one infection control nurse and SSI rates were produced for selected operations and surgical services. The program was conducted in general accordance with the 1999 HICPAC guidelines, but differed in surveillance strategy. Operations were limited to two to three risk classifications, assigned by the infection control nurse. RESULTS: The overall SSI rate was 4.5%, with 2.4% in clean surgery. Apart from increases in the 3rd, 4th, 13th, and 14th years, rates remained relatively stable during the 16 years. Few significant decreases in SSI rates in surgical services or specific operations were shown, apart from the following: vascular surgery, 8.1% to 5% between the first 8 years and the last 8 years; general surgery services, 9% to 5%, and gynecology, 15.8% to 1.7%, both in the first year compared with in subsequent years; and gastric operations, 21% to 4.3% between the first year and the second year. Organ/space infection was identified in 0.5% of 17,804 operations, including 0.4% meningitis after neurosurgical procedures, 3% graft infections after vascular bypass operations, and 0.2% intra-abdominal infections after abdominal surgery. CONCLUSIONS: With the use of basic principles of surveillance and modest resources, procedure specific SSI rates were produced, with little significant change during the 16 years. Despite limitations in case-finding, risk stratification, feedback, and surveillance methods, the overall SSI rates were comparable with other published data. PMID- 11868892 TI - Masking of neutropenic patients on transport from hospital rooms is associated with a decrease in nosocomial aspergillosis during construction. AB - To prevent nosocomial pulmonary aspergillosis during hospital construction, neutropenic patients with hematologic malignancy were required to wear high efficiency masks when leaving their rooms. The rate of nosocomial aspergillosis decreased from 0.73 per 1,000 hospital patient-days during fiscal years 1993 to 1996 to 0.24 per 1,000 hospital patient-days during fiscal years 1996 to 1999 (P < .001). High-efficiency masks reduced nosocomial aspergillosis during hospital construction. PMID- 11868893 TI - Failure of oral antimicrobial agents in eradicating gastrointestinal colonization with vancomycin-resistant enterococci. AB - Eradication of gastrointestinal colonization with vancomycin-resistant enterococci (VRE) using a combination of oral bacitracin and gentamicin was evaluated. Twenty-eight evaluable treated patients were matched with 28 control patients. After 3 months of follow-up, 5 patients (18%) in the treatment group versus 1 patient (4%) in the control group (P = .2) had negative results on stool cultures, with a similar frequency of VRE bacteremia (P = .8). The use of oral bacitracin plus gentamicin did not reduce VRE colonization or bacteremia. PMID- 11868894 TI - Sensor-operated faucets: a possible source of nosocomial infection? AB - Recently, contamination of sensor-operated faucets (SOFs) with Pseudomonas aeruginosa was observed. To evaluate odds ratios, we conducted a case-control study in which handle-operated faucets served as controls. No statistically significant difference in P. aeruginosa counts was observed between SOFs and regular faucets in our study (odds ratio, 0.0; 95% confidence interval, 0.0 to 39.0; two-sided P exact = .99). PMID- 11868895 TI - Healthcare epidemiology is the paradigm for patient safety. PMID- 11868896 TI - An outbreak of extended-spectrum beta-lactamase-producing Klebsiella species in a neonatal intensive care unit in Brazil. PMID- 11868897 TI - Factors of skin ageing share common mechanisms. AB - Ageing has been defined as the accumulation of molecular modifications which manifest as macroscopic clinical changes. Human skin, unique among mammalians insofar as it is deprived of fur, is particularly sensitive to environmental stress. Major environmental factors have been recognized to induce modifications of the morphological and biophysical properties of the skin. Metabolites from ingested or inhaled substances do affect skin, which is also sensitive to endogenous hormone levels. Factors as diverse as ultraviolet radiation, atmospheric pollution, wounds, infections, traumatisms, anoxya, cigarette smoke, and hormonal status have a role in increasing the rate of accumulation of molecular modifications and have thus been termed 'factors of ageing'. All these factors share as a common feature, the capability to directly or indirectly induce one of the steps of the micro-inflammatory cycle, which includes the expression of ICAM-1 in endothelial cells. This triggers a process leading to the accumulation of damages in the skin resulting in skin ageing since ICAM-1 expression provokes recruitment and diapedesis of circulating immune cells, which digest the extracellular matrix (ECM) by secreting collagenases, myeloperoxidases and reactive oxygen species. The activation of these lytic processes provokes random damage to resident cells, which in turn secrete prostaglandines and leukotrienes. These signaling molecules induce the degranulation of resident mast cells which release the autacoid histamine and the cytokine TNF-alpha thus activating endothelial cells lining adjacent capillaries which release P-selectin and synthesize ICAM-1. This closes a self-maintained micro-inflammatory cycle, which results in the accumulation of ECM damage, i.e. skin aging. In this paper we review the evidence that two factors able to induce macroscopical and molecular modifications in the skin, protein glycation and stretch, activate the micro-inflammatory cycle. We further present evidence that three additional factors, two external factors (electromagnetic fields and psychological stressors) and one internal factor (neuropeptides) also activate the micro inflammatory cycles and may therefore be considered as factors of skin ageing. PMID- 11868898 TI - Mitochondrial alterations, cellular response to oxidative stress and defective degradation of proteins in aging. AB - Respiratory function decline and increase of oxidative stress in mitochondria have been proposed as important contributors to human aging. A wide spectrum of alterations in aged individuals and senescent cells are similar and are correlated to cellular response to sublethal dose of oxidative stress. These alterations and responses include: (1) decline in mitochondrial respiratory function; (2) increase in the rate of production of reactive oxygen species (ROS); (3) accumulation of mitochondrial DNA (mtDNA) mutations; (4) increase in the levels of oxidative damage to DNA, protein, and lipids; and (5) decrease in the capacities of degradation of oxidatively damaged proteins and other macromolecules. Responses to oxidative stress and their subsequent interactions in tissues result in the deleterious effect of ROS on the cellular function, which culminate in aging and degenerative diseases. In this review, we focus on the roles that ROS play in age-related oxidative damage to mtDNA and proteins and oxidative stress responses at the molecular and cellular levels. The alterations of gene expression profiles elicited by oxidative stress in aging animals are discussed. We suggest that the increase in mitochondrial production of ROS and decline in the cellular capacity to cope with oxidative stress and subsequent accumulation of mtDNA mutations and oxidized proteins play an important role in the aging process. PMID- 11868899 TI - Modulation of hepatic and renal glucocorticoid receptors during aging of mice. AB - The regulation of hepatic and renal glucocorticoid receptors (GRs) in young (4 weeks) and senescent (120 weeks) mice was studied. Significant changes in the level of GRs from liver and kidney were detected, whereas the affinity (Kd) for the hormone [3H]dexamethasone did not change in young and old mice. The concentration of GRs was markedly decreased in liver (25%) and kidney (33%) of old mice as compared to young ones. The magnitude of heat activation of GR complexes was more pronounced in both the tissues at young age compared to old. In addition, we have found changes in the heat activation-inhibition studies of GRs, using polyunsaturated fatty acids (PUFAs) as measured by binding to DNA cellulose and purified nuclei. Linoleic acid (C18:2) exhibited significant decrease in heat activation of both hepatic and renal hormone-bound GR, with higher magnitude of inhibition in young liver (64%) and kidney (68%) as compared to old (41% and 43%, respectively). Arachidonic acid (C20:4) was also found to be an inhibitor of activation causing significant decrease in hepatic GR activation, with greater inhibition in young liver with respect to old, however, without any such difference in the kidney of young and old mice. Furthermore, DNase I digestion and extraction of nuclear-bound GR-complexes from both the tissues reveal lesser extraction in old liver (26%) and kidney (24%) compared to young tissues, indicating chromatin condensation with aging, thereby controlling the accessibility to such transcription factors as GRs. These findings indicate that the changes in the GR concentration, activation-modulation by PUFAs and chromatin organization, that take place during aging, may contribute to functional changes in glucocorticoid action mechanisms in senescent animals. PMID- 11868900 TI - Chromosome and aging: genetic conception of aging. AB - The mutation level (chromosome aberrations covering telomere regions), modifications of chromosome structure (level of condensed chromatin identified by the methods of electron microscopy and differential scanning microcalorimetry; level of C-banding constitutive heterochromatin; transcriptional activity of DNA dependent RNA polymeraze; Ag-positive NORs and associations of acrocentric chromosomes) and reparation (intensity of unscheduled DNA synthesis and the frequency of sister chromatid exchanges) have been studied in lymphocyte cultures from individuals at the age of 72-114 to reveal the chromosome functional organization at late stages of ontogenesis and to find explanations of some senile pathologies. The analysis of obtained results showed: 1. Chromosome progressive heterochromatinization (condensation of eu- and heterochromatin regions) occurs at aging; 2. Decrease of repair processes and increase in frequency of chromosome aberrations in aging are secondary to the progressive heterochromatinization. Chromosome heterochromatinization is a key factor of aging; 3. Chromosome heterochromatinization may be the reason for some senile pathologies; 4. Chromosome heterochromatinization is an area where one should seek the ways for prolonging the lifespan. PMID- 11868901 TI - DNA double-strand breaks in mouse kidney cells with age. AB - A Biojector device fitted with a CO2 cartridge was used to prepare single cell suspensions from kidneys of 12-month- (middle-aged) and 24-month-old (old) C57B1/6 mice. Microgel electrophoresis of DNA from these cells revealed a modest but significant 7.3% increase (P = 0.04) in DNA double-strand breaks in old mice. This increase is equivalent to the DNA damage induced by 0.1 Gray of X-rays (5 double-strand breaks) in kidney cells of 10-month-old mice, as determined by a standard calibration curve. Greater DNA damage with aging was also positively correlated with higher levels of pathology in the kidneys. PMID- 11868902 TI - Neural-specific overexpression of drosophila plenty of SH3s (DPOSH) extends the longevity of adult flies. AB - Extended longevity mutants are extremely useful to understand the molecular mechanism of longevity determination. Here we report identification and characterization of the Drosophila Plenty of SH3s (DPOSH) gene, a candidate that might be associated with the extended longevity phenotype. DPOSH encodes a protein containing a RING finger domain and four SH3 domains. We showed that neural-specific overexpression of DPOSH could extend the mean longevity of adult flies by 14% at 25 degrees C without affecting viability or morphology. In contrast, forced expression of DPOSH in developing imaginal discs produced various phenotypes including lethality and morphological defects such as loss of crossvein, notched wing, and disordered hair polarity. Puckered, a target gene of JNK/SAPK pathway, was activated by overexpression of DPOSH and the forced expression phenotypes were suppressed by introducing a mutation of Drosophila JNK (bsk) or JNKK (hep), suggesting that the JNK/SAPK signaling pathway is one of the critical elements in the determination of longevity. PMID- 11868903 TI - "I no longer believe that cell death is programmed. ..", an interview with Vincent Cristofalo. PMID- 11868904 TI - Effect of ondansetron on the contractile responses to positive inotropic agents in electrically driven left atria of rats. AB - We studied the interaction of ondansetron with positive inotropic agents, including serotonin (5-hydroxytryptophane, 5-HT), noradrenaline, 4-aminopyridine (4-AP), calcium chloride, or with reserpine in isolated electrically driven rat atria. Concentrations of 5-HT ranging from 1 to 64 microg/ml increased atrial contractions in a dose-dependent manner. The inotropic effect of 5-HT in the right atria appeared to be weaker than that in the left atria. Ondansetron (30 microg/ml) depressed the positive inotropic effect of lower 5-HT concentrations . This effect was thought to be due to the local anesthetic action of ondansetron or its agonistic interaction with inhibitory imidazoline receptors on the sympathetic nerve endings that reduce noradrenaline release. The positive inotropic effect of 5-HT was abolished almost completely by cyproheptadine (2 microg/ml) and was reversed only partially by pretreatment with reserpine (1 or 3 mg/kg). This result was considered as evidence for the participation of pre- and post-junctional 5-HT2A receptors and the involvement of the sympathetic nervous system in the positive inotropic action of 5-HT in rat atria. Experiments with atropine (1 microg/ml) in atria from reserpine-pretreated rats revealed that the parasympathetic component of the autonomic nervous system is not involved in the inotropic action of 5-HT. Ondansetron (30 microg/ml) tended to increase the positive inotropic effects of noradrenaline, 4-AP, and calcium chloride, which were partially significant at certain concentrations. This result might be due to the activation of both pre- and post-junctional 5-HT2A receptors or due to the inhibition of noradrenaline reuptake into the sympathetic nerve endings through the activation of imidazoline receptors. From these findings, we conclude that the positive inotropic effect of 5-HT in the electrically driven rat atria seems to be mediated primarily by its interaction with 5-HT2A receptors, which are likely to be found on the pre- and post-junctional structures. Other mechanisms that might be involved in this relation are also discussed. PMID- 11868906 TI - Effect of cefaperazone/sulbactam and ampicillin/sulbactam on the in vitro activity of human erythrocyte glucose-6-phosphate dehydrogenase. AB - We investigated the effects of the antibiotic drugs cefaperazone/sulbactam and ampicillin/sulbactam on the in vitro activity of the enzyme glucose-6-phosphate dehydrogenase (G6PD). The enzyme was purified from human erythocytes using 2',5' ADP-Sepharose 4B affinity gel. The enzymatic activity was measured spectrophotometrically at 340 nm, according to the method of Beutler. The I50 values were determined from Activity % - [Drug] graphs, and the Ki constants and inhibition types for each drug were determined using Lineweaver-Burk graphs. The I50 value was 13.5 mg/ml for cefaperazone/sulbactam and 36 mg/ml for ampicillin/sulbactam. The Ki constants were 10.16 for ampicillin/sulbactam and 38.22 for cefaperazone/sulbactam. Cefaperazone/sulbactam competitively inhibited G6PD activity, whereas ampicillin/sulbactam non-competitively inhibited the activity of the enzyme. PMID- 11868905 TI - Renovascular actions of adenosine in the isolated perfused rat kidney: possible underlying mechanisms. AB - We studied the renovascular action of adenosine on isolated perfused rat 10 min after drug injections. Adenosine was applied intraarterially as a single bolus injection in logarithmically increasing doses (0.3-30 microg). Adenosine treatment induced a biphasic vascular-response, namely, an initial vasoconstriction followed by a long-lasting vasodilation. Pretreatment with 0.1. 0.3, or 1.0 mM theophylline or quinidine (2 microg/ml) significantly depressed both components of the adenosine response. The vasoconstrictor response to adenosine was not affected by either 0.5 or 1.0 microg/ml dihydroergocristine. whereas the vasodilatory response was dose-dependently reduced. The biphasic response to adenosine was markedly depressed by 10 microg/ml indomethacin and was augmented by combining this agent with quinidine. We studied the possible roles of the platelet activating factor (PAF) and nitric oxide-cGMP systems in the renovascular actions of adenosine. Tebokan (a PAF antagonist) antagonized both components of the response, but methylene blue (MM) reduced only the pressory part Electron-microscopic examination of kidneys exposed for 15 min to MM showed some acute degenerative alterations and constriction in the glomeruli. From these findings, we conclude that the P1/A1, and P2x purinoceptors, the prostaglandins, PAF, and the NO-cGMP systems have a share in the renovascular actions of adenosine. PMID- 11868907 TI - Changes in effective and lethal doses of intravenous anesthetics and lidocaine when used in combination in mice. AB - We studied the interactions between a local anesthetic agent, lidocaine, and two general anesthetic drugs, propofol and ketamine, in mice. We used two end points: hypnosis, reflected by loss of the righting reflex, and death. The ED50 for hypnosis and the LD50 were determined for each drug separately, and a dose response curve was prepared for each drug, using combinations of propofol lidocane and ketamine-lidocaine at three different dose ratios. Probit and isobolographic analyses revealed supra-additive (synergistic) interactions between lidocaine and each of the other anesthetic agents regarding both the effective dose and the lethal dose. No significant difference was found between propofol and ketamine regarding the supraadditive effect. PMID- 11868908 TI - Simvastatin induces apoptosis of cultured rat cardiomyocytes. AB - Considering the therapeutic effect of statins (3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors) and simvastatin in patients with coronary heart disease, our first hypothesis was that simvastatin should inhibit apoptosis (programmed cell death) in angiotensin II-treated cultured myocytes. But after realizing that simvastatin stimulates apoptosis, we changed our hypothesis and began to study its apoptotic effect in primary cultured rat cardiomyocytes. We found that simvastatin induced apoptosis in a dose-dependent manner (0.1 to 3 micromol/L), as evidenced by the appearance of increased DNA fragmentation in agarose gels and characteristic apoptotic patterns in nuclei labeled with Hoechst 33342, as well as increased activity of caspase 3. FACS analysis of simvastatin treated cardiomyocytes showing annexin V binding and propidium iodide exclusion ruled out the possibility of necrosis. Increased intracellular enzymatic activity of creatine phosphokinase, aldolase, and lactic dehydrogenase, markers for normal cell function, could reflect the hypertrophic effect of simvastatin. The results indicate that simvastatin-induced apoptosis in cultured heart cells is concentration-dependent and additive to the apoptotic effect of angiotensin II. PMID- 11868909 TI - Using a Euclid distance discriminant method to find protein coding genes in the yeast genome. AB - The Euclid distance discriminant method is used to find protein coding genes in the yeast genome, based on the single nucleotide frequencies at three codon positions in the ORFs. The method is extremely simple and may be extended to find genes in prokaryotic genomes or eukaryotic genomes with less introns. Six-fold cross-validation tests have demonstrated that the accuracy of the algorithm is better than 93%. Based on this, it is found that the total number of protein coding genes in the yeast genome is less than or equal to 5579 only, about 3.8 7.0% less than 5800-6000, which is currently widely accepted. The base compositions at three codon positions are analyzed in details using a graphic method. The result shows that the preference codons adopted by yeast genes are of the RGW type, where R, G and W indicate the bases of purine, non-G and A/T, whereas the 'codons' in the intergenic sequences are of the form NNN, where N denotes any base. This fact constitutes the basis of the algorithm to distinguish between coding and non-coding ORFs in the yeast genome. The names of putative non coding ORFs are listed here in detail. PMID- 11868910 TI - Post Hartree-Fock and density functional theory studies on structure and conformational stability of nitrosoethylene and substituted compounds of nitrosoethylene. AB - Post Hartree-Fock and density functional theory (DFT) methods were used to study the different conformers of nitrosoethylene H-CH=CH-NO, and substituted compounds of the nitrosoethylene R-CH=CH-NO (R = Cl, NH2, N(CH3)2, OH, OCH3). The molecules were optimized at MP2/6-31G* level of theory of ab initio and B3LYP/6-31G* and B3PW91/6-31G* levels of theory of DFT. Special emphasis has been given to the effect of substitution of pi-electron donor groups NH2, N(CH3)2, OH, and OCH3, which play a major role in modifying the geometrical parameters of -N=O group by the electronic transmission effects through the central group -CH=CH-. The ability of DFT methods to predict the bond length adjacent to the atoms having lone pair electrons has been discussed. The conformational stabilities have been studied using the relative energies and DFT parameters such as chemical hardness and chemical potential. The role of intra-molecular hydrogen bond on the equilibrium structure has been discussed. The vibrational spectra for the different conformers of the nitrosoethylene and substituted compounds have been generated using the MP2/6-31G* level of theory. PMID- 11868911 TI - Novel atom-type AI indices for QSPR studies of alcohols. AB - The novel vertex degree v(m) for heteroatom in molecular graph is derived on the basis of the valence connectivity delta(v) of Kier-Hall. The newly proposed atom type Al indices and previously proposed Xu index, are further modified for compounds with heteroatoms by replacing the vertex-degree of heteroatom by the proposed v(m). The multiple linear regression using the modified Xu index and Al indices can provide high-quality QSPR models for the normal boiling points (BP), molar volumes (MV), molar refractions (MR), and molecular total surface areas (TSA) of alcohols with up to 17 non-hydrogen atoms. The results imply that these physical properties may be expressed as a linear combination of the individual indices related to molecular size and atom-types. For each of the four properties, the correlation coefficient r is greater than 0.996 and particularly the decrease in the standard error is within the range of 61 83% compared with the simple linear models based on the modified Xu index, and the standard errors are 3.814, 0.939, 0.187, and 3.348 for BP, MV, MR, and TSA, respectively. The final models correspond to a fit error of 2.33, 0.70, 0.53, and 0.95% for BP, MV, MR, and TSA, respectively. The more general leave-n-out method is used to do the cross-validation. The cross-validation demonstrates the outstanding predictive power of the final models. The contributions of individual indices are used to illustrate the role of the molecular size and individual groups in molecules. The results indicate that physical properties of alcohols are dominated by the molecular size. On the other hand, although the hydrogen-bonding interactions caused by the -OH group play an important role in determining the normal BPs, the branching seems to be a more important factor influencing the MVs, MRs, and TSAs than the hydrogen-bonding interaction. The contribution of individual atom type or group to properties is not a constant and depends on its structural environment in a molecule. PMID- 11868912 TI - QSPR models derived for the kinetic data of the gas-phase homolysis of the carbon methyl bond. AB - A quantitative structure-property relationship study was carried out on the kinetic parameters of the gas-phase homolysis for 58 different C-CH3 bonds using the CODESSA program. Six-parameter models were developed for the prediction of the log k (1047 K) and the parameters of the Arrhenius equation, log A and E. These correlations were obtained by employing the theoretical molecular descriptors, derived from only the information encoded in the chemical structure of compounds. PMID- 11868913 TI - Development of a quantitative structure-property relationship model for predicting the electrophoretic mobilities. AB - Electrophoretic mobility (mu0) is the most important parameter governing the separation of solutes in capillary zone electrophoresis. In this paper, a new model was constructed by means of a multilayer neural network using extended delta-bar-delta (EDBD) algorithm to estimate complex property of electrophoretic mobilities of aliphatic carboxylates and amines from simpler experimental properties. The molecular weight (W), molecular volume (V), the code (+1 or -1) of acid and base and pK value were used as input parameters to predict electrophoretic mobility. The networks' architecture and the learning times were optimized. The optimum artificial neural networks (ANNs) could give excellent prediction results. PMID- 11868914 TI - Automation of a procedure to find the polynomial which best fits (kappa, c1, c2, T) data of electrolyte solutions by non-linear regression analysis using MATHEMATICA software. AB - A MATHEMATICA package, 'CONDU.M', has been developed to find the polynomial in concentration and temperature which best fits conductimetric data of the type (kappa, c, T) or (kappa, c1, c2, T) of electrolyte solutions (kappa: specific conductivity; ci: concentration of component i; T: temperature). In addition, an interface, 'TKONDU', has been written in the TCL/Tk language to facilitate the use of CONDU.M by an operator not familiarised with MATHEMATICA. All this software is available on line (UPV/EHU, 2001). 'CONDU.M' has been programmed to: (i) select the optimum grade in c1 and/or c2; (ii) compare models with linear or quadratic terms in temperature; (iii) calculate the set of adjustable parameters which best fits data; (iv) simplify the model by elimination of 'a priori' included adjustable parameters which after the regression analysis result in low statistical significance; (v) facilitate the location of outlier data by graphical analysis of the residuals; and (vi) provide quantitative statistical information on the quality of the fit, allowing a critical comparison among different models. Due to the multiple options offered the software allows testing different conductivity models in a short time, even if a large set of conductivity data is being considered simultaneously. Then, the user can choose the best model making use of the graphical and statistical information provided in the output file. Although the program has been initially designed to treat conductimetric data, it can be also applied for processing data with similar structure, e.g. (P, c, T) or (P, c1, c2, T), being P any appropriate transport, physical or thermodynamic property. PMID- 11868915 TI - A graph-theoretic method to identify candidate mechanisms for deriving the rate law of a catalytic reaction. AB - Stoichiometrically, exact candidate pathways or mechanisms for deriving the rate law of a catalytic or complex reaction can be determined through the synthesis of networks of plausible elementary reactions constituting such pathways. A rigorous algorithmic method is proposed for executing this synthesis, which is exceedingly convoluted due to its combinatorial complexity. Such a method for synthesizing networks of reaction pathways follows the general framework of a highly exacting combinatorial method established by us for process-network synthesis. It is based on the unique graph-representation in terms of P-graphs, a set of axioms, and a group of combinatorial algorithms. In the method, the inclusion or exclusion of a step of each elementary reaction in the mechanism of concern hinges on the general combinatorial properties of feasible reaction networks. The decisions are facilitated by solving linear programming problems comprising a set of mass balance constraints to determine the existence or absence of any feasible solution. The search is accelerated further by exploiting the inferences of preceding decisions, thereby eliminating redundancy. As a result, all feasible independent reaction networks, i.e. pathways, are generated only once; the pathways violating any first principle of either stoichiometry or thermodynamics are eliminated. The method is also capable of generating those combinations of independent pathways directly, which are not microscopically reversible. The efficiency and efficacy of the method are demonstrated with the identification of the feasible mechanisms of ammonia synthesis involving as many as 14 known elementary reactions. PMID- 11868916 TI - Prediction of protein structural classes by support vector machines. AB - In this paper, we apply a new machine learning method which is called support vector machine to approach the prediction of protein structural class. The support vector machine method is performed based on the database derived from SCOP which is based upon domains of known structure and the evolutionary relationships and the principles that govern their 3D structure. As a result, high rates of both self-consistency and jackknife test are obtained. This indicates that the structural class of a protein inconsiderably correlated with its amino and composition, and the support vector machine can be referred as a powerful computational tool for predicting the structural classes of proteins. PMID- 11868917 TI - Positive association between HLA-DRB1*07 and specific IgE responses to purified major allergens of D. pteronyssinus (Der p 1 and Der p 2). PMID- 11868918 TI - Is hypersensitivity pneumonitis important? PMID- 11868919 TI - Monoclonal anti-IgE antibody: a novel therapy for allergic airways disease. AB - LEARNING OBJECTIVES: To familiarize the practitioner with a novel (monoclonal anti-immunoglobulin [Ig]E antibody) form of therapy for allergic airways disease. To understand the relevance of IgE as a therapeutic target. To appreciate the concepts behind the design of the molecule. To learn how anti-IgE was used in clinical trials. To anticipate the likely effects (efficacy and safety) in clinical use in patients with allergic asthma and with allergic rhinitis. To characterize the types of patients who might benefit from this therapy. DATA SOURCES: Published data for preclinical and clinical studies. RESULTS: Omalizumab is a nonimmunogenic, nonanaphylactogenic monoclonal anti-IgE antibody. In clinical use, omalizumab reduces levels of serum-free IgE. Given subcutaneously in patients with moderate-severe allergic asthma, omalizumab reduced exacerbations compared with placebo, and at the same time it allowed inhaled corticosteroids to be reduced or withdrawn. In patients with allergic rhinitis, omalizumab reduced the severity of symptoms and rescue antihistamine usage versus placebo. In both settings, quality of life was improved with active treatment relative to placebo. The drug seems safe and well tolerated. CONCLUSION: As the first clinical anti-IgE agent, omalizumab is an interesting new addition to the currently available therapies for allergic airways disease. The benefits demonstrated underline the importance of IgE in these conditions. The use of anti IgE in other IgE-mediated allergic diseases warrants further research. PMID- 11868920 TI - Recurrent periorbital cellulitis and otitis media in an asthmatic child with chronic diarrhea and short stature. PMID- 11868921 TI - Positive association between HLA-DRB1*07 and specific IgE responses to purified major allergens of D. pteronyssinus (Der p 1 and Der p 2). AB - BACKGROUND AND OBJECTIVE: Human leukocyte antigen (HLA) class II gene products are involved in the antigen presentation of exogenous antigens. The aim of this study was to evaluate whether specific immunoglobulin (Ig)E responses to purified major allergens of Dermatophagoides pteronyssinus (Der p 1 and Der p 2) were associated with genotypes of HLA-DRB1 alleles. SUBJECTS AND METHODS: Peripheral venous blood samples were collected from two groups of unrelated Korean adolescents: 168 subjects with positive skin response (wheal > or = 3 mm) to crude D. pteronyssinus allergens and 94 age-matched controls with negative skin response and low serum-specific IgE to crude D. pteronyssinus allergens. The former group was found to contain 104 with high serum-specific IgE to crude D. pteronyssinus allergens, 100 with high serum-specific IgE to Der p 1, and 122 with high serum-specific IgE to Der p 2. Genotypes of the HLA-DRB1 alleles were determined using polymerase chain reaction-sequence specific oligonucleotide probes. RESULTS: HLA-DRB1*07 was significantly higher in subjects with a high serum-specific IgE response to crude D. pteronyssinus allergens than the controls (16.3% vs 3.2%, odds ratio [OR] = 5.93, corrected P [Pc] = 0.02). The excess of DRB1*07 was more marked in those with high serum-specific IgE responses to Der p 1 or Der p 2 than the controls (20.0% vs 3.2%, OR = 7.58, Pc = 0.004; 18.9% vs 3.2%, OR = 7.05, Pc = 0.006). Among subjects with high serum IgE to Der p 1, DRB1*13 significantly increased than the controls (32.0% vs 13.8%, OR = 2.93, Pc = 0.03). CONCLUSIONS: We clearly observed the association between HLA-DRB1 alleles and specific IgE responsiveness to D. pteronyssinus major allergens. The molecular mechanism of HLA-DRB1*07 and DRB1*13 involvement in D. pteronyssinus specific IgE responsiveness awaits further investigation. PMID- 11868922 TI - The long-term outcome in acute, subacute, and chronic forms of pigeon breeder's disease hypersensitivity pneumonitis. AB - BACKGROUND: Pigeon breeder's disease (PBD) is an avian-induced hypersensitivity pneumonitis. The limited data that exist on the long-term outcome for the disease are conflicting. OBJECTIVE: We sought to determine the long-term outcome of patients with PBD to evaluate the course of the disease. METHODS: Review a case series of 18 pigeon breeders that were followed from 4 to 26 years after initial diagnosis. RESULTS: Fourteen breeders reduced their bird contact significantly; four kept their birds. In eight patients with the acute form of the disease, pulmonary function either did not change from normal or improved to normal. Of five patients with subacute disease, four had persistent symptoms. All had mildly abnormal spirometry with either restrictive or obstructive changes which persisted. Five patients with chronic disease exhibited daily dyspnea. They had severe abnormalities of lung function at diagnosis. Of these patients, three improved and two deteriorated. Chest radiographs were of little value in predicting outcome. CONCLUSIONS: Patients with the acute form of PBD have the best prognosis, compared with those with subacute and chronic forms. Although some improvement could be seen in most individuals, recovery may still not be complete. Those with the chronic form are at particular risk for morbidity. PMID- 11868923 TI - Prevalence of food allergies in young adults and their relationship to asthma, nasal allergies, and eczema. AB - BACKGROUND: The true prevalence of food allergy in adults is generally thought to be uncommon. It is unknown whether there are any relationships between food allergy and atopic diseases. OBJECTIVE: To determine the prevalence of probable immunoglobulin (Ig)E-mediated food allergy to peanut, shrimp, cow's milk, wheat, and egg as defined by a positive skin prick test result and relevant clinical history to the same food, and to explore the relationship with atopic diseases. METHODS: Cross-sectional epidemiologic study. One thousand one hundred forty-one randomly selected young adults (aged 20 to 45 years) underwent skin prick testing to five common food allergens (cow's milk, peanut, egg white, shrimp, and wheat), completed a detailed questionnaire, including validated items on respiratory symptoms, history of asthma and other allergic conditions, as well as undergoing lung function testing. RESULTS: Just over one percent (1.3%, n = 15) had probable IgE-mediated food allergy. The prevalence of probable IgE food allergy was: <0.27% for wheat, 0.09% (95% confidence interval = 0.0 to 0.49%) each for cow's milk and egg, 0.53% (0.21 to 1.09%) for shrimp, and 0.61% (0.25 to 1.26%) for peanut. Those with probable IgE peanut and shrimp allergy were significantly more likely to have current asthma and doctor-diagnosed asthma. Wheeze and history of eczema were also associated with peanut allergy, whereas nasal allergies were associated with shrimp allergy. CONCLUSIONS: The prevalence of probable IgE mediated food reactions is rare in young adults. Some positive associations between probable IgE-mediated food allergy and allergic diseases were found, but larger study numbers are required to confirm these results. PMID- 11868925 TI - Effect of ingestion of honey on symptoms of rhinoconjunctivitis. AB - BACKGROUND: Allergic rhinoconjunctivitis is a common disorder, affecting >20% of people of all socioeconomic strata. Despite this high prevalence, relatively few sufferers seek professional medical help, presumably because of a widespread reliance on complementary remedies. OBJECTIVE: We investigated the widely held belief among allergy-sufferers that regular ingestion of honey ameliorates the symptoms of allergic rhinoconjunctivitis. METHODS: The study was conducted at the University of Connecticut Health Center's Lowell P. Weicker General Clinical Research Center. Thirty-six participants who complained of allergic rhinoconjunctivitis were recruited. All recruits were scratch-tested at entry for common aeroallergens. The cohort was randomly assigned to one of three groups, with one receiving locally collected, unpasteurized, unfiltered honey, the second nationally collected, filtered, and pasteurized honey, and the third, corn syrup with synthetic honey flavoring. They were asked to consume one tablespoonful a day of the honey or substitute and to follow their usual standard care for the management of their symptoms. All participants were instructed to maintain a diary tracking 10 subjective allergy symptoms, and noting the days on which their symptoms were severe enough to require their usual antiallergy medication. RESULTS: Neither honey group experienced relief from their symptoms in excess of that seen in the placebo group. CONCLUSIONS: This study does not confirm the widely held belief that honey relieves the symptoms of allergic rhinoconjunctivitis. PMID- 11868924 TI - A double-blind, randomized, single-dose, crossover comparison of levocetirizine with ebastine, fexofenadine, loratadine, mizolastine, and placebo: suppression of histamine-induced wheal-and-flare response during 24 hours in healthy male subjects. AB - BACKGROUND: Levocetirizine is the active enantiomer of cetirizine, a potent drug with little metabolism widely used for allergic rhinitis and urticaria. OBJECTIVE: This study compares the potency, consistency, onset, and duration of action of levocetirizine with other popular antihistamines. METHODS: Levocetirizine 5 mg, ebastine 10 mg, fexofenadine 180 mg, loratadine 10 mg, mizolastine 10 mg, or placebo in single doses were given to 18 healthy male volunteers in a double-blind, crossover, randomized fashion. Wheal-and-flare responses to epicutaneous histamine dihydrochloride (100 mg/mL) challenge were measured at 0, 0.5, 1, 2, 4, 6, 8, 10, 12, and 24 hours after each dose. RESULTS: The overall effect of each drug was evaluated by the area under the curve (0 to 24 hours). Levocetirizine was the most potent and consistently effective drug for inhibiting the histamine-induced wheal-and-flare surface areas. Ebastine, fexofenadine, and mizolastine ranked next and had almost identical effects for inhibiting the wheal. Loratadine was the least potent drug. Levocetirizine, fexofenadine, and mizolastine inhibited the wheal-and-flare response after 1 hour and reached their peak for inhibition after 4 hours. Ebastine and loratadine could be distinguished from placebo only after 4 hours. After treatment with levocetirizine, all 18 subjects had >95% inhibition of the wheal response at one timepoint. Fexofenadine, mizolastine, and ebastine were inhibitory in declining order. All treatments were considered safe and well tolerated. CONCLUSIONS: Levocetirizine, the active enantiomer of cetirizine, is more potent and consistent than other popular H1 antihistamines for blocking the cutaneous response to histamine. These findings may predict the efficacy of this drug in treating allergic disorders. PMID- 11868926 TI - In vitro deposition of fluticasone aerosol from a metered-dose inhaler with and without two common valved holding chambers. AB - BACKGROUND: Previous in vitro aerosol deposition experiments indicate that the corticosteroid respirable dose from a metered-dose inhaler (MDI) can vary by threefold depending on the specific valved holding chamber (VHC) MDI combination. OBJECTIVE: We compared in vitro aerosol deposition from a fluticasone propionate MDI (Flovent, GlaxoSmithKline, Research Triangle Park, NC) to that of the same MDI used in combination with two VHCs (EasiVent, Dey, Napa, Ca; and AeroChamber Plus, Monaghan Medical Corp, Plattsburgh, NY) to evaluate how these VHCs affect the respirable dose of fluticasone. METHODS: The respirable dose (aerosol particles 1 to 5 microm in size) of fluticasone was determined by sampling 5 x 110-microg actuations from each configuration (MDI alone, MDI plus AeroChamber Plus, and MDI plus EasiVent) in multiples of ten using a well established, in vitro cascade impactor method. Fluticasone aerosol was washed from individual impactor stages with 50% methanol and quantified via ultraviolet high-pressure liquid chromatography. Differences among outcomes were determined using analysis of variance. RESULTS: Mean respirable dose from AeroChamber-Plus (47.9 +/- 6.9 microg/actuation) was not different (P > 0.05) from that produced by the MDI alone (50.3 +/- 2.2 microg/actuation). EasiVent respirable dose (27.0 +/- 3.6 microg/actuation) was less than that produced by either the AeroChamber-Plus or the MDI alone (P < 0.001). CONCLUSIONS: VHCs do not display equivalent in vitro performance with a fluticasone MDI. If a patient needs a VHC, clinicians should use available in vitro performance information to aid in selecting the best device. PMID- 11868927 TI - Inter-rater evaluation of a clinical scoring system in children with asthma. AB - BACKGROUND: Many clinical scores that measure the degree of asthma are used without adequate evaluation of inter-rater reliability. When reliability is tested, most often the Cohen K statistic is used, which limits the comparative results of only two raters at a time. OBJECTIVE: To evaluate inter-rater agreement of a clinical asthma score using a multi-rater K statistic. METHODS: Four raters administered a clinical asthma score to 17 children with clinical asthma. Five items were evaluated: O2 requirement, inspiratory breath sounds, accessory muscle use, expiratory wheeze, and cerebral function. For each, a score of zero indicated a normal state; one, moderate impairment; two, severe impairment. A multi-rater kappa statistic was used as a measure of agreement among all four raters simultaneously. This was applied using hand calculations then cross-checked by using a standard statistical syntax, a component of the Statistical Package for Social Sciences (SPSS 9.0). RESULTS: Application of the multi-rater K statistic revealed strong agreement among raters on oxygenation (K = 0.759), moderate agreement for expiratory wheeze and cerebral function (K = 0.698), and poor agreement for accessory muscle use (K = 0.528) and inspiratory breath sounds (K = 0.316). CONCLUSIONS: The level of agreement varied by item with the least subjective item, O2 requirement, demonstrating the highest inter rater correlation. A multi-rater kappa statistic can be applied to data obtained from a clinical scoring instrument either manually or by using statistical syntax provided by SPSS. PMID- 11868928 TI - Pulmonary function changes and immunomodulation of Th 2 cytokine expression induced by aminophylline after sensitization and allergen challenge in brown Norway rats. AB - BACKGROUND AND OBJECTIVE: Evidence has shown that aminophylline has bronchoprotective, anti-inflammatory, and immunomodulatory effects. Our purpose was to evaluate the effect of different doses of aminophylline on the late-phase reaction, bronchial hyperresponsiveness (BHR) and T cell-related cytokine mRNA expression in brown Norway rats induced by ovalbumin (OA) sensitization. METHODS: Forty rats were equally divided into four groups. Groups I, II, and III animals were sensitized and subsequently provoked with OA. Aminophylline 25 mg/kg was given intraperitoneally to the group I animals and 5 mg/kg to group II animals. Group III animals received intraperitoneal normal saline. Group IV breathed aerosolized saline as a control. After OA provocation, the animals were anesthetized. Pulmonary function tests were performed at baseline and after varying doses of acetylcholine. Thereafter, bronchoalveolar lavage was performed and the lungs were examined histologically. Total RNA was extracted from lung tissue and reverse transcriptase-polymerase chain reaction was performed using primers for interleukin (IL)-2, IL-4, IL-5, IL-6, IL-10, interferon-gamma, inducible nitric oxide synthase, and beta-actin. RESULTS: Group III had worse pulmonary function tests, more severe BHR, and more severe lung inflammation, higher IL-4 and IL-10 cytokine levels in bronchoalveolar lavage fluid, and higher IL-4, IL-5, IL-6, IL-10, tumor necrosis factor-alpha and inducible nitric oxide synthase mRNA expression than the other three groups. Expression of IL-2 and interferon-gamma was significantly reduced in group III. CONCLUSIONS: Both low and high dose aminophylline are effective in preventing late-phase bronchoconstriction, BHR, and an inflammatory response. Aminopylline decreases T helper cell 2-related cytokine mRNA expression but increases T helper cell 1 related cytokines mRNA expression. PMID- 11868929 TI - Correlation between presumed sinusitis-induced pain and paranasal sinus computed tomographic findings. AB - BACKGROUND: The correlation between facial and/or head pain in patients clinically suspected of having sinusitis and actual localized findings on sinus computed tomographic (CT) imaging are poorly understood. OBJECTIVE: To prospectively evaluate the relationship of paranasal sinus pain symptoms with CT imaging. METHODS: Two hundred consecutive patients referred by otolaryngologists and internists for CT of the paranasal sinuses participated by completing a questionnaire immediately before undergoing CT. Three radiologists blinded to the patients' responses scored the degree of air/fluid level, mucosal thickening, bony reaction, and mucus retention cysts using a graded scale of severity (0 to 3 points). The osteomeatal complexes and nasolacrimal ducts were also evaluated for patency. Bivariate analysis was performed to evaluate the relationship between patients' localized symptoms and CT findings in the respective sinus. RESULTS: One hundred sixty-three patients (82%) reported having some form of facial pain or headache. The right temple/forehead was the most frequently reported region of maximal pain. On CT imaging the maxillary sinus was the most frequently involved sinus. Bivariate analysis failed to show any relationship between patient symptoms and findings on CT. Patients with a normal CT reported a mean 5.88 sites of facial or head pain versus 5.45 sites for patients with an abnormal CT. CONCLUSIONS: Patient-based responses of sinonasal pain symptoms fail to correlate with findings in the respective sinuses. CT should therefore be reserved for delineating the anatomy and degree of sinus disease before surgical intervention. PMID- 11868931 TI - Maples, genus Acer. PMID- 11868930 TI - Fluticasone propionate/salmeterol combination compared with montelukast for the treatment of persistent asthma. AB - BACKGROUND: Asthma is a chronic disease characterized by inflammation and bronchoconstriction. Medications that are able to effectively treat both components are advantageous. OBJECTIVE: To compare the efficacy of an inhaled corticosteroid and a long-acting beta2-agonist combination product with a leukotriene antagonist for initial maintenance therapy in patients who were symptomatic while receiving short-acting beta2-agonists alone. METHODS: A 12 week, randomized, double-blind, double-dummy, multicenter study was conducted in 432 patients 15 years of age and older with persistent asthma who were symptomatic on short-acting beta2-agonists alone. Fluticasone propionate 100 microg and salmeterol 50 microg combination product (FSC) twice daily or montelukast 10 mg once daily was administered. RESULTS: At endpoint, compared with montelukast, FSC significantly increased morning predose forced expiratory volume in 1 second (0.61 +/- 0.03 L vs 0.32 +/- 0.03 L), morning peak expiratory flow rate (peak expiratory flow rate; 81.4 +/- 5.9 L/minute vs 41.9 +/- 4.8 L/minute), evening peak expiratory flow rate (64.6 +/- 5.3 L/minute vs 38.8 +/- 4.7 L/minute), the percentage of symptom-free days (40.3 +/- 2.9% vs 27.0 +/- 2.7%), the percentage of rescue-free days (53.4 +/- 2.8% vs 26.7 +/- 2.5%), and the percentage of nights with no awakenings (29.8 +/- 2.5% vs 19.6 +/- 2.1%) (P < or = 0.011, all comparisons). At endpoint, FSC significantly reduced asthma symptom scores (-1.0 +/- 0.1 vs -0.7 +/- 0.1) and rescue albuterol use (-3.6 +/- 0.2 puffs/day vs -2.2 +/- 0.2 puffs/day) compared with montelukast (P < 0.001). At endpoint, patients treated with FSC also had a significantly greater improvement in quality of life scores and were more satisfied with their treatment compared with montelukast-treated patients (P < or = 0.001). Both treatments were well tolerated. CONCLUSIONS: Initial maintenance therapy with FSC provides greater improvement in asthma control and patient satisfaction than montelukast. PMID- 11868936 TI - Meeting the challenge of finding resources for ophthalmic nurses on the World Wide Web. AB - The World Wide Web ("the Web") is a macrocosm of resources that can be overwhelming. Often the sheer volume of material available causes one to give up in despair before finding information of any use. The Web is such a popular resource that it cannot be ignored. Two of the biggest challenges to finding good information on the Web are knowing where to start and judging whether the information gathered is pertinent and credible. This article addresses these two challenges and introduces the reader to a variety of ophthalmology and vision science resources on the World Wide Web. PMID- 11868934 TI - Vaccine development against HIV-1: current perspectives and future directions. AB - The development of an efficacious vaccine against the human immunodeficiency virus (HIV) is of great urgency, because it is accepted that vaccination is the only means capable of controlling the AIDS pandemic. The foundation of HIV vaccine development is the analysis of immune responses during natural infection and the utilization of this knowledge for the development of protective immunization strategies. Initial vaccine development and experimentation are usually in animal models, including murine, feline, and nonhuman primates. Experimental vaccine candidates are closely studied for both efficacy and safety before proceeding to human clinical trials. There are a number of different therapeutic and prophylactic vaccine strategies currently being studied in human clinical trials. Vaccine strategies that are being tested, or have previously been tested, in humans include subunit, DNA plasmid, and viral vector, and combinations of these various strategies. Some of the results of these trials are promising, and additional research has focused on the development of appropriate chemical and genetic adjuvants as well as methods of vaccine delivery to improve the host immune response. This review summarizes the vaccine strategies that have been tested in both animal models and human clinical trials. PMID- 11868937 TI - Finding a nursing role in the prevention of eye injuries in the construction trades. AB - Construction workers have mostly preventable eye injuries. This pilot study investigated a convenience sample of 75 (72 men, 3 women) apprentice operating engineers. Most (52%) of the apprentices had been injured twice on the job. Injury was associated with age and longevity in the construction industry. There were no significant differences in health perceptions as measured by the Health Self-determinism Index when the safety glasses wearers were compared with non wearers of safety glasses (t[52] = -.94, P > .05) and the injured group was compared with the noninjured group (t[62] = -.38, P > .05). Literacy issues, recall, and underreporting of injuries may have affected the results. Nurses need to teach health promotion strategies to operating engineers. PMID- 11868939 TI - Bright light therapy and seasonal affective disorder. PMID- 11868938 TI - Diplopia: double the fun! Part 1: History taking. AB - The history is the most important part of a diploic evaluation. Determining the type of diplopia and its onset, duration, and frequency are critical questions that must be answered. The examiner must ask the right question to get the right answers. This article will review the history of the diploic patient step by step and give suggestions on how to avoid some common problems. Common causes for diplopia will also be reviewed. Knowing what questions to ask can make a big difference to you, the patient, and the ophthalmologist and turn a challenge into a rewarding experience. PMID- 11868932 TI - Experimental corneal allograft rejection. AB - The major findings regarding corneal allograft rejection in experimental animals are reviewed. The principal anatomic and biological feature of the cornea that determines the immunologic privilege of this tissue is its avascularity. The surgical trauma of transplantation compromises the immunologic privilege, putting corneal allografts at risk for immune rejection. During the past 50 yr, rabbits, rats, and mice have been used extensively in the study of the process of immunologically mediated corneal allograft rejection. It is clear that the inflammation and neovascularization of the graft that occurs following transplantation predisposes a corneal allograft to the classic cell-mediated immune rejection response. The antigenicity of cornea cells has been studied and has been found to be significantly lower compared to other cells and tissues. Rejection of acorneal allograft is acell-mediated process directed against major histocompatibility complex antigens involving both CD4+ T helper cells and CD8+ cytotoxic cells. The prevention of corneal allograft rejection depends on the development of topically applied compounds that can prevent inflammation and vascularization and inhibit the activation of T lymphocytes. Considerable progress has been made using immunomodulators, including blocking antibodies and soluble coreceptor blocking agents such as CTLA4-Ig. Combinations of antiangiogenic agents and immunomodulators hold great promise for preventing corneal allograft rejection in patients. PMID- 11868933 TI - Insights into the immunopathogenesis of multiple sclerosis. AB - Multiple sclerosis (MS) is a chronic inflammatory disease of the central nervous system (CNS). Significant progress has been made in our understanding of the etiology of MS. MS is widely believed to be an autoimmune disease that results from aberrant immune responses to CNS antigens. T cells are considered to be crucial in orchestrating an immunopathological cascade that results in damage to the myelin sheath. This review summarizes the currently available data supporting the idea that myelin reactive T cells are actively involved in the immunopathogenesis of MS. Some of the therapeutic strategies for MS are discussed with a focus on immunotherapies that aim to specifically target the myelin reactive T cells. PMID- 11868940 TI - Ocular histoplasmosis. AB - Presumed ocular histoplasmosis syndrome can be as benign as not knowing one has a vision problem or can result in shock and disbelief on losing central vision and dealing with legal blindness. PMID- 11868952 TI - A threat to the professionalism of nursing. PMID- 11868953 TI - Designing a learning-needs survey--10 steps to success. PMID- 11868935 TI - Interactions of integrins with their partner proteins in leukocyte membranes. AB - Integrins participate in many aspects of immunologic and inflammatory responses, especially those involving cell migration, adherence, and activation. Although leukocyte integrins such as complement receptor type 3 (CR3) are known to carry out certain functions without the intervention of other plasma membrane receptors, many plasma membrane proteins are now known to physically interact and functionally cooperate with integrins. Several of these interactions are highly dynamic within cell membranes; thus integrin-partner protein interactions change during certain physiological processes. This allows an extraordinary adaptability of the system to prime and promote proinflammatory signaling. Since our discovery of the CR3-FcgammaRIIIB interaction, the plasma membrane protein repertoire of beta1, beta2, and beta3 integrins has grown to include: FcgammaRIIA (CD32), uPAR (urokinase-type plasminogen activator receptor; CD87), CD14, voltage-gated K+channels (Kv1.3), integrin-associated protein (IAP), CD98, tetraspans (TM4SF), insulin receptors, and PDGFbeta receptors. In this article we will highlight certain features of this growing field of research, especially with regard to their relevance in immunology and inflammation. PMID- 11868954 TI - Searching for the final answer: factors contributing to medication administration errors. AB - Medication errors have reached epidemic proportions in the United States with proposed resolutions having little effect. Many errors go unreported because of nurses' fear of retaliation. Yet these unreported errors may contain the very information needed to uncover system flaws. Until all causes are examined and researched thoroughly, systems will continue to fail. These unknown factors contributing to medication errors will remain elusive unless health care providers are willing and able to focus attention where it is needed-on systems rather than individuals. Patients have enough fears when entering health care institutions; medication errors should not be one of them. PMID- 11868956 TI - Use of a search model to enhance patient education in a clinical setting. AB - Many paths lead toward the goal of safe, high-quality health care. One component of this goal is to obtain current and relevant information for client care. Patients, as well as health care providers, use the World Wide Web to find information. With the growing number of documents on the Web, an organized approach to searching fosters locating desired material, evaluating authenticity and accuracy, and applying the material to the patient, student, or practitioner population, as appropriate. This article explores the use of a search model in a nurse-managed clinic for homeless and lower income patients. Nurses in a wide variety of settings use the search process described to locate useful health care information. PMID- 11868955 TI - Antidote Dilemma--an activity to promote critical thinking. AB - BACKGROUND: In a health care environment of increased patient acuity and greater demands on nursing practitioners, the ability to think critically is requisite to functioning safely and proficiently. METHOD: Definitions and tenets of critical thinking promulgated by Brookfield (1987) and Paul (1992) provided the theoretical foundation for Antidote Dilemma, an activity designed to promote critical thinking. RESULTS: Nurses who participated in the activity identified plausible solutions to a nursing dilemma, considered context in identifying alternatives, and thought about their thinking. CONCLUSION: Critical thinking is most likely to occur and continue when it is supported by others and repeatedly practiced. Continuing/staff-development educators play a vital role in promoting critical thinking among nursing practitioners. PMID- 11868957 TI - Nutrition knowledge of nurses in long-term care facilities. AB - BACKGROUND: The purpose of this study was to determine knowledge-based nutrition competencies of nurses and describe needed nutrition education relative to reducing protein-calorie malnutrition (PCM) in nursing home residents. METHOD: A sample of 44 nurses (35 RNs, 9 LPNs) from five eastern Washington nursing homes completed a 50-item questionnaire that measured nutrition knowledge based on Benner's Novice to Expert Model. RESULTS: Nurses scored an average 65% +/- 11% on the nutrition examination. CONCLUSION: An educational program addressing the identified deficient areas could improve the ability of nursing staff to assess and monitor resident food intake, thereby reducing the prevalence of PCM of residents in nursing homes. PMID- 11868958 TI - Addressing sexuality-related needs in practice: perspectives of maternal/child and women's health nurses. AB - A quantitative, descriptive survey was conducted using Waterhouse's instrument, Survey of Sexuality-Related Nursing Practice. The purpose of this study was to examine the extent to which maternal/child and women's health nurses address sexuality in their practice and to assess the influence of select variables on that practice. A sample of maternal/child and women's health registered nurses (n = 130) was systematically selected from the 1995 Association of Women's Health, Obstetric and Neonatal Nurses' (AWHONN) District VII mailing list. Findings reveal incongruities in maternal/child and women's health nurses' perspectives and the incorporation of sexuality-related nursing interventions into practice. PMID- 11868959 TI - Problems in the outcomes of nursing education create challenges for continuing education. AB - BACKGROUND: The purpose of this study was to identify the challenges for continuing education by evaluating nurses' perceptions of quality of learning outcomes among graduates of nursing programs in Finland. The evaluation occurred 5 years after graduation from a bachelor's program, which means that learning outcomes can be weighed against the practice of nursing. METHOD: The data were collected in 1996 by postal questionnaires, which were divided into four main domains--basic nursing skills, interactive skills, counseling skills, and collaborative skills. The sample consisted of 522 graduating nursing students. The response rate was 68% (n = 354). RESULTS: The respondents felt most confident with their basic nursing, collaborative, and counseling skills. They felt least confident with their interactive skills, particularly with patients with alcohol or drug addiction problems. CONCLUSION: The results highlight some of the strengths and weaknesses of the nurse education system and provide insights for continuing education. PMID- 11868960 TI - Cervical cancer screening among South Asian women in Canada: the role of education and acculturation. AB - Cervical cancer screening by Papanicolaou (Pap) smear lowers the incidence and provides early detection of cervical cancer and is a preventative health care measure that should be available on a regular basis to all women at risk. As the population of ethnic women increases, it is important to assess whether these women are aware of and are utilizing this health service. A self-administered questionnaire was completed by 124 South Asian women, aged 18 to 60 years. Sixty two South Asian students were selected from a university setting and 62 Tamil women were selected from a community center for South Asian women. This study examined the knowledge and use of Pap smears in South Asian women in Canada, and whether their level of acculturation or formal education influenced whether they know about and receive Pap smears. Low level of knowledge about the Pap test and a low prevalence of Pap testing behaviour was significantly correlated (p < 0.001) with a low level of formal education, education taking place outside of Canada, and a low index of acculturation. South Asian students were significantly more acculturated than Tamil women (p < 0.001). In conclusion, there was a low level of knowledge and prevalence of cervical cancer screening among South Asian women. This study highlights the need for educational interventions in Canada directed to ethnic women especially from South Asia. PMID- 11868961 TI - Women living with ovarian cancer: dealing with an early death. AB - Our intent to explore the emotion of anger in women living with ovarian cancer revealed a basic social concern of "dealing with an early death." The findings of this grounded theory study also identified the core variable of "finding meaning in life" as assisting the nine women in dealing with an early death. The categories of hope and physical wellness influenced the women's search for meaning after a diagnosis of ovarian cancer. Several action and interactional coping strategies were identified as positive coping behaviors. The consequences of finding meaning in life was a perception of well-being defined by the women as satisfaction with their lives. Conversely, an inability to find meaning in life resulted in feelings of despair. The women's perceptions of well-being were not static and could fluctuate as they lived with ovarian cancer. Implications for further research and practice are discussed. PMID- 11868962 TI - Information women receive about heart attacks and how it affects their knowledge, beliefs, and intentions to act in a cardiac emergency. AB - In this study we investigate (a) what information women receive on acute myocardial infarction (AMI) from different information sources, (b) how women evaluate this information, and (c) how information acquisition is related to women's knowledge of AMI, personal risk perceptions, and behavioral intentions regarding a cardiac emergency. A random sample of 844 women 50 years and over were telephoned in the state of Washington. Repeated measures regression analyses showed that women receive most of their information on AMI from the mass media, even though they evaluated the media as less believable, less useful, less clear, and making less of an impression than information received from health care providers. Logistic regression analyses showed that the total number of information sources from which heart attack information was received in the past year was positively related to knowledge of AMI symptoms, beliefs about AMI, and personal risk perceptions. The results are discussed in terms of their pragmatic implications. PMID- 11868963 TI - Social factors that make South African women vulnerable to HIV infection. AB - The degree to which women are able to control various aspects of their sexual lives is clearly a critical question for health promotion and the prevention of AIDS. It is evident that social factors such as the high rate of rape, the unfavourable economic position of women, and the inability to insist on condom usage make South African women unable to negotiate the timing of sex and the conditions under which it occurs. They are thus rendered powerless to protect themselves against HIV infection. Prevention campaigns often do not take into account the reality of the daily lives of South African women and the difficulties they face gaining control over their own sexual lives. The rampant spread of this disease can only be stemmed if the subordinate position of women is acknowledged and addressed. PMID- 11868964 TI - Cultural and gender issues related to HIV/AIDS prevention in rural Swaziland: a focus group analysis. AB - HIV/AIDS is a serious public health problem in Swaziland, a small land-locked Southern African country. The epidemic affects all subpopulations, but women are increasingly at risk for contracting the disease. Focus groups were conducted in a rural area to obtain qualitative information on the rural dimensions of HIV/AIDS, vulnerability to HIV/AIDS, and sociocultural factors influencing the spread of AIDS. Five themes were identified from analyses of the focus groups that are relevant in developing an AIDS prevention program for this population in Swaziland. Implications for future research and health care practice focus on gender-sensitive cultural research (e.g., women and men's roles and responsibilities in rural Swaziland society) and how social, economic, and cultural factors impact the spread of AIDS. Recommendations include reorienting and allocating resources for health, education, and social services to address the relative neglect of rural areas and strengthening policies and programs to achieve the equal participation of all women in all aspects of society's decisions. Specifically, policies related to economic and food security should result in programs to improve local access by women to all resources. PMID- 11868965 TI - Health care services for lesbian and bisexual women: some Canadian data. AB - Specific health needs and health services for lesbian and bisexual women are relatively invisible in existing research, with little Canadian data on these topics. To redress such gaps, data were selectively analyzed from a larger study with 98 lesbian/bisexual women living in a Maritime Province in Canada. As part of a semistructured interview, women were asked to indicate whether a particular health care service was important to them. General physical examination, Pap smear, breast examination, and holistic medicine were the four services chosen with the highest frequency. HIV/AIDS information/screening and safer sex information were judged important by over half of the participants. When asked to rank their top three services, however, participants chose the physical examination, holistic medicine, and psychological counseling. Our results are discussed in terms of barriers to basic health care, accessibility of services, and the importance of accurate information about AIDS and safer sex. PMID- 11868966 TI - Physical health and preventive health behaviors among Thai women in Brisbane, Australia. AB - In this article we describe the health status of Thai migrant women in Australia. Women's physical health, their use of health services, and adherence to preventive health behaviors are presented. The results are based on data from a descriptive study of 139 women living in Brisbane who completed a structured questionnaire. This information was supplemented with data from in-depth interviews with 17 of the women to gain additional information and insight into immigration, language, social support, and the use of health services. Although the majority of women reported being in good health and only a small percentage suffered from chronic diseases, inherent problems can influence their future health status. These include women's low participation in preventive health programs, unhealthy life style and behavior, perceptions of health services, availability of social support during illness, and issues related to care from general practitioners. Women's understanding of health education and information was influenced by their language skills. Language difficulties and the perceived high costs of medical care in Australia were contrasted with the lower cost and perceived efficiency and efficacy of Thai medical practitioners, and as a result some women delayed medical attention until home visits to Thailand. PMID- 11868967 TI - Grandmothers and women's health: from fragmentation to coherence. AB - There is a paucity of models that drive integrated research agendas, and coherent approaches to development and progress of knowledge about women's health. In this article, we review four major models of women's health, present conditions supporting more integrative and coherent models of women's health based on the recommendations by two international conferences, and address major paradoxes inherent in women's health areas. For integrative and coherent models of women's health, we propose to incorporate visions and insights of previous models in developing a more coherent model that includes three major components integration, transition, and marginalization. PMID- 11868968 TI - Penetration of diltiazem into breast milk and its pharmacokinetics in the lactating rabbit. AB - 1. The aim was to investigate the milk transfer and pharmacokinetics of diltiazem (DTZ) in the lactating rabbit following DTZ intravenous (i.v.) administration. In addition, DTZ metabolism in mammary tissue and milk was also studied. 2. The pharmacokinetic parameters that largely determine drug disposition (AUC, VD, CL) showed no significant differences between the non-lactating and lactating rabbit. 3. When DTZ was administered to the lactating rabbit, the observed DTZ milk-to blood AUC ratio (M/B) closely correlated with the calculated ratio, as predicted by a diffusional model, suggesting that DTZ passes into milk via non-ionic diffusion and that other factors which may affect the milk transfer seem to have limited relevance. 4. After a single intravenous dose of DTZ to the lactating rabbit, deacetyldiltiazem (M1) and demethyldiltiazem (MA) were observed in blood, but only M1 could be detected in milk. 5. In conclusion, DTZ seems to diffuse freely into milk after its i.v. administration to the lactating rabbit and should not be given to nursing mammals because of the potential risk for their young. This risk may be even higher because of the presence of M1 (a pharmacologically active metabolite) in milk after administration of the parent drug. PMID- 11868969 TI - Metabolism of 10,11-dihydro-10-hydroxyimino-5H-dibenz/b, f/azepine-5-carboxamide, a potent anti-epileptic drug. AB - 1. 10,11-Dihydro-10-hydroxyimino-5H-dibenzo/b, f/azepine-5-carboxamide (BIA 2 024) is a new anti-epileptic drug similar to oxcarbazepine (OXC) in structure and efficacy, but with a preferred pharmacodynamic profile. It possesses high in vitro activity, but since oximes are usually metabolized to their corresponding ketones, it is important to know whether its is vivo potency is a result of acting as a prodrug of OXC or if it is acting on its own. 2. The drug was given orally to rats, mice and rabbits, the metabolites identified and pharmacokinetic profiles compared between those species. Furthermore, the pharmacokinetic profile of the main metabolite was established in the rat. The results were compared to in vitro metabolism studies with liver microsomes from different mammalian species and humans. 3. In an atypical reaction for oximes, BIA 2-024 in rats was rapidly (t(max) = 2h) metabolized to the non-active 10-nitro-derivative (BIA 2 254), whereas rabbits and particularly mice oxidized the oxime moiety to a much lower extent. BIA 2-254 was then transformed to OXC and subsequently to the 10 hydroxy derivative and other minor metabolites. 4. In vitro data showed a very similar cross-species behaviour as the in vivo results; human liver microsomes catalysed the oxidation of BIA 2-024 to the nitro metabolite only at a low rate, and the same was observed for the subsequent metabolism to OXC. 5. The results allow prediction of the in vivo metabolism of BIA 2-024 in humans, where this drug is most likely absorbed efficiently and excreted mainly as the parent compound with a relatively low hepatic clearance. With the exception of rat, BIA 2-024 does not act as a prodrug of OXC. PMID- 11868970 TI - LC-MS-MS analysis of hydromorphone and hydromorphone metabolites with application to a pharmacokinetic study in the male Sprague-Dawley rat. AB - 1. A high performance liquid chromatography-mass spectrometry-mass spectrometry (LC-MS-MS) assay was developed for the analysis of hydromorphone and its metabolites, namely dihydromorphine, dihydroisomorphine, hydromorphone-3 glucuronide, dihydromorphine-3-glucuronide and dihydroisomorphine-3-glucuronide, in rat plasma samples. 2. Analytes were extracted by solid-phase extraction using C2 cartridges. The extraction recoveries were > 76% for all analytes. Both intra- and interassay variabilities were < or = 12%. Using a plasma sample size of 100 microl, the limits of detection were 7.0 nmol(-1) (2.0 ng ml(-1)) for hydromorphone, dihydromorphine and dihydroisomorphine and 11 nmol l(-1) (5.0 ng ml l(-1)) for hydrormorphone-3-glucuronide, dihydromorphine-3-glucuronide and dihydroisomorhine-3-glucuronide at a signal-to-noise ratio = 3. 3. The present assay was applied to a pharmacokinetic study in rat after intraperitoneal administration of hydromorphone. PMID- 11868971 TI - Absorption, metabolism and excretion of 4-chloro-2-methylphenoxyacetic acid (MCPA) in rat and dog. AB - 1. The oral no overall adverse effect level (NOAEL) for chronic toxicity of 4 chloro-2-methylphenoxyacetic acid (MCPA) in rat is approximately 1.3 mg kg(-1) and in dog is 0.2 mg kg(-1). In an attempt to explain the difference in toxicology between these species, rats and dogs were orally dosed with (14C)-MCPA at 5 or 100 mgkg(-1) and plasma toxicokinetics, rates and routes of excretion and biotransformation were investigated. 2. Elimination of radioactivity in rat plasma was biphasic and in dog was monophasic. Rat eliminated radioactivity from plasma significantly faster than dog (approximate values biased on total radioactivity: 5 mg kg(-1) rat: t 1/2 dist 3.5 h, t 1/2 elim 17.2-36.2 h, AUC(0 infinity) 230 microg equiv hg(-1); 5 mg kg(-1) dog: t 1/2 47h, AUC(0-infinity) 2,500 microg equiv h g(-1); 100 mg kg(-1) rat: t 1/2 dist 10h, t 1/2 elim 10.27 25.4h, AUC(0-infinity) 5,400 microg equiv hg(-1); l00 mg kg(-1) dog: t 1/2 h, AUC(0-infinity) 20,500 microg eqiv h g(-1). 3. For both species, the principal route of excretion was in urine but renal elimination was notably more rapid and more extensive in rat. 4. In both rat and dog, excretion of radioactivity was mainly as MCPA and its hydroxylated metabolite hydroxymethylphenoxyacetic acid (HMCPA). In rat, both were mainly excreted as the free acids although a small proportion was conjugated. In dog, the proportion of HMCPA was increased and the majority of both species was excreted as glycine or taurine conjugates. 5. These data, along with previously published accounts, indicate that renal elimination of MCPA in dog is substantially slower than in rat resulting in disproportionate elevation of AUC (based on total radioactivity) in dog compared with rat. PMID- 11868972 TI - Species differences in the metabolism of the antitumour agent 5,6 dimethylxanthenone-4-acetic acid in vitro: implications for prediction of metabolic interactions in vivo. AB - 1. Mouse studies have indicated that the antitumour effects of 5,6 dimethylxanthenone-4-acetic acid (DMXAA) are dramatically potentiated in combination with other drugs, and it has been proposed that optimization of the therapeutic potential of DMXAA would exploit combination therapy. The aim was to identify the most appropriate animal model for further investigations of the pharmacokinetics of possible DMXAA-drug combinations and their extrapolation to patients. 2. Qualitatively, the metabolic profile for DMXAA in liver microsomes was similar in mouse, rat, rabbit and humans, with glucuronidation and 6 methylhydroxylation the two major metabolic pathways. In all species, the intrinsic clearance by glucuronidation was at least 2-fold that due to hydroxylation. There was significant variability in the in vitro kinetic parameters (Km, Vmax), with the mouse being the least efficient DMXAA metabolizer compared with the other species. 3. Mouse, rat and rabbit renal microsomes exhibited DMXAA glucuronidation activity, but only the rabbit showed 6 methylhydroxylation. For the total in vitro CL(int) (Vmax/Km) by glucuronidation and 6-methylhydroxylation, the ratio of kidney:liver was 0.67, 0.03 and 0.34 in the mouse, rat and rabbit respectively. However, taking into account the liver and kidney weight difference, it is apparent that the in vivo renal metabolism would not be a major contributor to the overall elimination of DMXAA. 4. The inhibitory profile for liver DMXAA glucuronidation was similar across species, but there was remarkable interspecies variability in the inhibition of liver DMXAA 6methylhydroxylation. 5. Extrapolation of in vitro intrinsic clearance to in vivo gave a significant underestimation of plasma clearance for all species. However, there was a significant allometric relationship for plasma clearance and volume of distribution, but not for maximum tolerated dose across species. 6. The results indicate that animal models may have a limited role in the extrapolation to patients of drug interactions with agents such as DMXAA that have immunomodulating activity that may vary widely between species. PMID- 11868974 TI - Photo quiz. Piloleiomyomas. PMID- 11868973 TI - Tuberous sclerosis. AB - Tuberous sclerosis (TS) is an autosomal dominant disorder with a significant range of clinical expression. Renal, pulmonary, central nervous system, and cardiac complications may result in severe morbidity. Early recognition of the syndrome is important to ensure prompt identification of systemic involvement. We review the clinical features of tuberous sclerosis and discuss recent advances in our understanding of its pathogenesis and appropriate management. PMID- 11868975 TI - Acquired cutis laxa associated with multiple myeloma. AB - Cutis laxa is an uncommon condition characterized by loose and redundant skin. Biopsy results are positive for a reduction in or an absence of elastic fibers in the dermis. Cutis laxa is acquired or congenital. The acquired form is either a generalized insidious form (type I) or a form associated with prior inflammation (type II). Cardiovascular, pulmonary, gastrointestinal, and urologic complications may occur. In the past, cutis laxa was associated with plasma cell dyscrasia. We report on a characteristic cause of cutis laxa to alert clinicians to this uncommon manifestation of multiple myeloma. PMID- 11868976 TI - Extracorporeal photochemotherapy: a case report and update. AB - Extracorporeal photochemotherapy (ECP) was developed at Columbia Presbyterian Medical Center in the early 1980s for the treatment of cutaneous T-cell lymphoma (CTCL). ECP is now used primarily in the treatment of that disease at more than 100 centers worldwide. It also has been shown to be potentially effective in treating several autoimmune diseases. Most recently, it has been used in reversing solid-organ transplant rejection and graft-versus-host disease following bone-marrow transplantation. In this article, we present the case of one of the first patients treated with ECP and give an update on the current status of this therapy. PMID- 11868977 TI - Cutaneous consequences of photodynamic therapy. AB - Photodynamic therapy (PDT) has several cutaneous complications: photosensitivity is well known, but the other complications are rarely reported, Since late 1997, we have studied the dermatologic complications of using porfimer sodium PDT to treat either Barrett esophagus with high-grade dysplasia or gastroesophageal cancer in 72 consecutive patients. Cutaneous complications of PDT included serious phototoxicity requiring oral corticosteroid treatment (22 patients; 31%), herpes zoster (HZ) requiring hospitalization and intravenous antiviral treatment (1 patient; 1%), and erythema multiforme drug reaction related to porfimer sodium (1 patient; 1%). PDT-associated dermatologic complications were common and were not related to cutaneous photosensitivity. PMID- 11868978 TI - Metameric motor paresis following abdominal herpes zoster. AB - Motor neuropathy is an uncommon complication that may follow an outbreak of herpes zoster (HZ). About half of the reported cases have involved the cranial nerves, typically the facial nerve. The remaining cases have affected the nerves of the extremities. Interestingly, motor weakness of the thoracic segments is strikingly rare, even though this is where HZ most frequently occurs. The dermatologic literature reports only exceptions to this occurence. We report a new case of motor paresis following HZ infection in an abdominal location, where this complication can be easily misdiagnosed as abdominal herniation. PMID- 11868979 TI - Lichen aureus. AB - Lichen aureus is a localized variant of pigmented purpuric dermatitis (PPD) and is rarely reported. A case of lichen aureus in a 28-year-old woman and a 13-year old boy are presented. Biopsy results revealed the characteristic pattern of lichen aureus, including diffuse deposition of hemosiderin and dermal infiltrate of lymphocytes and histiocytes, some of which had Birbeck granules in the cytoplasm. The former case is unusual for its zosteriform distribution and localization (lower abdomen). PMID- 11868980 TI - The middle ground. PMID- 11868981 TI - Topical treatment of tinea capitis in a neonate. AB - Tinea capitis is the most common fungal skin infection in children. Given that this infection invades the hair shaft and the pilosebaceous unit, systemic antifungal therapy is the gold standard of treatment. Despite the neonate's increased susceptibility to infections, tinea capitis is rare in this population. We present the case of a 16-day-old infant with tinea capitis caused by Microsporum canis and effectively treated with topical bifonazole 1%. PMID- 11868982 TI - What's eating you? Loxosceles reclusa (brown recluse spider). PMID- 11868983 TI - Acute hemorrhagic pellagra in an Albanian refugee. AB - We report a peculiar case of hemorrhagic pellagra in an exhausted Albanian refugee who had walked for 3 days under sunny skies on his way from his country to Greece. The peculiarities of the case are the fulminant course of the disorder; the "terrifying" appearance of the patient (initially he was admitted to an emergency unit); the gangrenous appearance of the hemorrhagic lesions of the palms and fingernails; the disturbed hepatic function that gradually returned to normal; and the absence of a history of alcohol consumption, alcohol malabsorption, or drug intake. PMID- 11868984 TI - The dermatologist's role in diagnosing a rare disease--male breast cancer. AB - After discussing the rarity of male breast carcinoma, I present the case of a 75 year-old white man who, during an office visit for a second opinion regarding his eczema, was found to have breast cancer. (While examining the patient's integument, I found an inverted nipple and a subareolar mass.) I also discuss the need for dermatologists to emphasize to their patients the importance of complete general skin examinations. This case is, I believe, the first case of invasive primary male breast carcinoma to be reported in the dermatologic literature. PMID- 11868986 TI - Unconventional rules of small nuclear RNA transcription and cap modification in trypanosomatids. AB - This review focuses on the spliced leader (SL) RNA and uridylic acid-rich small nuclear RNAs (U-snRNAs) involved in pre-mRNA processing in trypanosomatid protozoa, with particular emphasis on the mechanism of transcription and cap formation. The SL RNA plays a central role in mRNA biogenesis by providing the unique cap 4 structure to the 5' end of all mRNAs by trans-splicing. The trimethylguanosine capped U-snRNAs, on the other hand, represent an unusual example among eukaryotic snRNAs in that they are transcribed by RNA polymerase III. This implies the existence of a distinctive mechanism for capping enzyme selection by the transcriptional machinery. Furthermore, the transcription units of U-snRNA genes offer yet another example of the variety of choices that have been established during eukaryotic evolution, namely that an upstream tRNA gene or tRNA-like gene provides extragenic promoter elements for a downstream small RNA gene. PMID- 11868985 TI - Small nucleolar RNAs: versatile trans-acting molecules of ancient evolutionary origin. AB - The small nucleolar RNAs (snoRNAs) are an abundant class of trans-acting RNAs that function in ribosome biogenesis in the eukaryotic nucleolus. Elegant work has revealed that most known snoRNAs guide modification of pre-ribosomal RNA (pre rRNA) by base pairing near target sites. Other snoRNAs are involved in cleavage of pre-rRNA by mechanisms that have not yet been detailed. Moreover, our appreciation of the cellular roles of the snoRNAs is expanding with new evidence that snoRNAs also target modification of small nuclear RNAs and messenger RNAs. Many snoRNAs are produced by unorthodox modes of biogenesis including salvage from introns of pre-mRNAs. The recent discovery that homologs of snoRNAs as well as associated proteins exist in the domain Archaea indicates that the RNA-guided RNA modification system is of ancient evolutionary origin. In addition, it has become clear that the RNA component of vertebrate telomerase (an enzyme implicated in cancer and cellular senescence) is related to snoRNAs. During its evolution, vertebrate telomerase RNA appears to have co-opted a snoRNA domain that is essential for the function of telomerase RNA in vivo. The unique properties of snoRNAs are now being harnessed for basic research and therapeutic applications. PMID- 11868987 TI - La protein and its associated small nuclear and nucleolar precursor RNAs. AB - After transcription by RNA polymerase (pol) III, nascent Pol III transcripts pass through RNA processing, modification, and transport machineries as part of their posttranscriptional maturation process. The first factor to interact with Pol III transcripts is La protein, which binds principally via its conserved N-terminal domain (NTD), to the UUU-OH motif that results from transcription termination. This review includes a sequence Logo of the most conserved region of La and its refined modeling as an RNA recognition motif (RRM). La protects RNAs from 3' exonucleolytic digestion and also contributes to their nuclear retention. The variety of modifications found on La-associated RNAs is reviewed in detail and considered in the contexts of how La may bind the termini of structured RNAs without interfering with recognition by modification enzymes, and its ability to chaperone RNAs through multiple parts of their maturation pathways. The CTD of human La recognizes the 5' end region of nascent RNA in a manner that is sensitive to serine 366 phosphorylation. Although the CTD can control pre-tRNA cleavage by RNase P, a rate-limiting step in tRNASerUGA maturation, the extent to which it acts in the maturation pathway(s) of other transcripts is unknown but considered here. Evidence that a fraction of La resides in the nucleolus together with recent findings that several Pol III transcripts pass through the nucleolus is also reviewed. An imminent goal is to understand how the bipartite RNA binding, intracellular trafficking, and signal transduction activities of La are integrated with the maturation pathways of the various RNAs with which it associates. PMID- 11868991 TI - Bye bye chemical disinfection with canisters. AB - This paper describes and evaluates disinfection techniques of dialysis monitors available in our dialysis unit. We demonstrate disinfection techniques with regards to the risk of human error. To achieve a high hygienic standard and to support the safety and efficiency of each disinfection procedure, we emphasise the use of single-use-disinfection cartridges. PMID- 11868992 TI - Hepatitis C virus: overview of clinical and technical perspectives. AB - The increasing incidence/prevalence of hepatitis C virus (HCV) infection in dialysis patients is a cause for concern. In particular, the problem of possible transmission of HCV within a dialysis unit needs to be investigated from both a clinical and technical perspective and substantial evidence generated to support future management. Nosocomial transmission has been repeatedly recognised as a major factor in the spread of HCV. Professional negligence and patient naivety can result in the rapid development of nosocomial infection and research has identified that the incidence of transmission, in environments where parental routes are accessible, is higher. There have been a limited number of investigations into transmission through dialysis equipment and to date this evidence seems to indicate that HCV RNA (cells) do not pass through the dialysis membranes. With so little evidence, there is a need for further clarity. Currently it appears that there is no general agreement about preventative measures. Despite a limited amount of evidence, there appears to be a wide variation in policies concerning the isolation of patients. This article will provide an overview of current clinical and technical research that addresses the management of this virus within the haemodialysis environment. It is also the intention of the author that the article acts as a catalyst to promote interest in an international collaborative research project (CRP), which will be launched by the EDTNA/ERCA Research Board. PMID- 11868989 TI - RNA-protein interactions that regulate pre-mRNA splicing. AB - Splicing of nuclear precursor messenger RNAs is an important and ubiquitous type of gene regulation in metazoans. Splicing joins the coding sequences called exons by removing the intervening noncoding sequences, introns, from primary transcripts. Alternative splicing generates an enormous repertoire of functional diversity by producing multiple RNAs and proteins from a single gene. In fact, recent genome sequences from several organisms suggest that splicing regulation is likely to provide an important source of functional diversity in more complex organisms. Because splice sites are short sequences at the ends of introns, the functional splice sites have to be distinguished from an excessively large number of sequences in the primary transcripts that resemble a splice site. Furthermore, alternative splice sites have to be correctly chosen at appropriate times. Thus, selection of proper splice sites remains a daunting biological problem. This review focuses on a few examples in which the molecular and biochemical basis for splice site selection is better understood. PMID- 11868990 TI - Practical initiatives in the prevention of cross infection. AB - In Belgium Infection Control Nurses must register postoperative wound infections, sepsis and ventilation associated pneumonia. At the author's hospital, the incidence of pressure sores is scored four times a year and there is a register of new patients with M.R.S.A. (methicillin resistant staphylococcus aureus) and tuberculosis. Procedures for hospital hygiene can be consulted in order to see which patients must be isolated and what kind of precautions must be taken. The microorganisms and diseases are sorted alphabetically and the cause of cross infection, isolation procedure and duration of isolation are noted. These procedures must be translated to the different departments and individual patients. For instance a patient with M.R.S.A. is strictly isolated in the general hospital, but not in the psychiatric department. As far as the haemodialysis unit is concerned, patients with chronic renal disease are more sensitive to infections. For this reason correct hand hygiene is very important. Hand washing, hand disinfection techniques and the use of gloves must be promoted. A microbiological control of the hands of staff once a year, combined with an educational programme, can motivate staff in a positive way Needle-stick injuries present a serious occupational hazard for health care workers, especially those working in a haemodialysis unit. Information and needleless haemodialysis may reduce the risk of needle-stick injuries and the risk of viral transmission. Can a nurse, at risk of viral contamination and transmission, refuse to treat a patient? Is the use of hats, overshoes, glasses necessary? How many times are these materials changed, etc.? This paper presents the data of 1. years of M.R.S.A. registration at the author's hospital. M.R.S.A. has become a serious problem in many hospitals since the mid 1970s. Strenuous efforts need to be taken to control its spread. Screening of the nose of patients and of staff can be helpful. The carriers can be treated with mupirocin ointment. Body washing with chlorhexidine is preferred. The use of vancomycin and teicoplanin is a decision for the physician. PMID- 11868988 TI - The 3' end formation in small RNAs. AB - Small RNAs are a major class of RNAs along with transfer RNAs, ribosomal RNAs, and messenger RNAs. They vary in size from less than 100 nucleotides to several thousand nucleotides and have been identified and characterized both in prokaryotes and eukaryotes. Small RNAs participate in a variety of cellular functions including regulating RNA synthesis, RNA processing, guiding modifications in RNA, and in transport of proteins. Small RNAs are generated by a series of posttranscriptional processing steps following transcription. While RNA 5' end structure, 5' cap formation, and RNA processing mechanisms have been fairly well characterized, the 3' end processing is poorly understood. Recent data point to an emerging theme in small RNAs metabolism in which the 3' end processing is mediated by the exosome, a large multienzyme complex. In addition to removal of nucleotides by the exosome, there is simultaneous rebuilding of the 3' end of some small RNA by adenylation and/or uridylation. This review presents a picture of both degradative and rebuilding reactions operative on the 3' end of some small RNA molecules in prokaryotes and eukaryotes. PMID- 11868993 TI - Self-monitoring of microbiological levels in dialysis fluids. AB - Self monitoring of dialysis fluids is important. The dialysis technician has experience of the microbiological situation of the fluid systems and can make more tests for personnel safety. The tests are not expensive and are very easy to learn. It seems that the cells, which generate endotoxins, cannot be determined by the plate count system. No tanks or permanent water flow UV sterilisation in the pipeline to the dialysis machines can prevent the build up of biofilms and higher endotoxin amounts in water. PMID- 11868994 TI - Microbiological survey of dialysate: vantage of use of sterile bag concentrate. AB - Haemodialysed patients are exposed to nearly 400 litres of dialysis water weekly. The bacterial contamination of treated dialysate and water induces acute pyrogenic reactions or chronic damage and cytokine activation. The aim of this study was to value the microbiological parameters of dialysis water and dialysate of our monitors by bacterial culture (measured as colony forming units [CFU]) of water samples at 37 degrees C after 48 hours, at 22 degrees C after 72 hours and after seven days, and by measuring endotoxin levels (endotoxin units [EU]). In our centre, there are 16 monitors (6 monitors use sterile dialysate fluid and 10 monitors use non sterile dialysate fluid). The chemicals used for disinfection are chlorine and paracetic acid. Water samples were taken under sterile procedures every three months for a year. No bacteria were found in the samples of water of the dialysis ring; EU were lower than the limit value of 0.25 EU/ml fixed by the European Pharmacopoeia. The concentration of CFU and EU of the dialysate, taken from monitors with a sterile bag, were lower than those of other monitors (p < 0.05 t Student test). However, the levels of CFU/ml and EU/ml of dialysate samples, taken from monitors with a non-sterile bag, were lower than the guideline value of the European Pharmacopoeia (v.n. CFU < 50 CFU/ml and EU < 0.05 EU/ml). Frequent examination of CFU and EU is essential to reduce the damage caused by the use of contaminated water, therefore the goal of future dialytic techniques will be the use of "sterile dialysate". PMID- 11868995 TI - Dialysis fluid contamination of pathways and life of microbes. AB - The fluid systems of a dialysis clinic are reviewed from a microbiological standpoint. Water, concentrate and dialysis fluid are the main fluids in the clinic. The quality of these fluids cannot be dealt with, without at the same time reviewing the systems delivering these fluids. The handling of fluids must be seen in a system perspective where every part is important. In the pretreatment of water before reverse osmosis, the incoming water determines the quality. After reverse osmosis,the maintenance in form of disinfection activities is decisive for the microbiological quality in the water system. The dialysis fluid quality is dependent on the water quality as it is produced at the end of the water system. It must be noted that it is not only the number of microorganisms that is of importance but also what the microorganisms do in the fluid systems. Microbiological analysis is not normally able to tell the complete microbiological quality of the fluid systems, as the inner surfaces where the microbial growth takes place are not sent to any laboratory. Consequently, what is seen in samples is only what can come off the surface. The only action that can prevent growth of microorganisms is disinfection but disinfection only is not the solution, it must also be performed regularly. Additionally, all areas of the fluid system must be disinfected. The principle of Quality Assurance including equipment, education and maintenance must be applied in order to ensure microbiological quality. PMID- 11868996 TI - Containers for waste causing peritonitis in CAPD patients. AB - Fungal infection is a rare cause of peritonitis in patients on chronic peritoneal dialysis. In this population, fungi are found in less than 2 percent of all cases of primary episodes of peritonitis (1). More often, a primary bacterial peritonitis treated with antibiotic therapy leads to secondary fungal infection (2). Candida species cause 74.5% of the episodes of fungal peritonitis (2). The fungi invade the peritoneal cavity from the skin peri- or intraluminally through the catheter (3). Filamentous fungi are rare (4,5). Treatment of fungal peritonitis commonly consists of removal of catheter and antifungal drugs (3). Here we describe two cases of fungal peritonitis caused by mycelial fungi, where the source of the microorganism could be special containers used for biological waste, which are popular in Germany PMID- 11868997 TI - Improving the nurse-patient relationship: a multi-faceted approach. AB - AIM: A Corporate Education Session was held to provide concrete strategies for overcoming several specific barriers in the daily nurse-patient relationship that negatively affect the patient, nurse, or both, and to provide the latest information about best practice as it affects nurses in renal care. METHODOLOGY: The session was led by a moderator who presented three video case studies to the audience. Communication strategies for recognising and overcoming nurse-patient communication barriers were presented. The audience expressed their views about each case study using voting pads, and a panel of experts addressed the comments of the audience and discussed guidelines for best practice in renal care. The panel comprised three experienced renal nurses, a senior nephrologist, and an expert in social interaction. CONCLUSIONS: Optimal treatment of patients with renal disease should include early treatment of anaemia, adequate levels of dialysis, and a multidisciplinary approach, responding to both the medical and the social needs of the patient. PMID- 11868998 TI - Care of the geriatric patient in chronic renal failure. AB - Currently, there are around one million chronic dialysis patients worldwide. More than half of these are older than 65. For various reasons this is a fast growing population. When the indication is considered to start dialysis, a distinction must be made between biological age in the third and fourth decade on the one hand, and a specific geriatric profile on the other, which implies adequate gerontological knowledge. Besides the patient-directed choice of the best tolerated treatment technique, the specific treatment of age-linked polypathology as well as the appropriate approach of inter- and intradialysis related problems regarding access and haemodynamics are the best guaranties of success in this highest age dialysis group. By means of a multidisciplinary geriatric assessment, which makes it possible to carry out an objective analysis and observation from the somatic, psychic and social points of view, an ultimate objective is achieved: the improvement of quality of life by means of rehabilitation. The survival of elderly dialysis patients has favourably evolved since the 90's after the introduction of EPO, bicarbonate dialysate, the monitoring of dialysis adequacy and support of prognostic and comorbid factors. The problems surrounding dialysis drop-out is a sensitive and controversial matter in which well founded communication with the patient, the family, the dialysis team, domestic care and the general practitioner is vital. "Add life to years and not years to life" remains a challenge for every nephrologist. PMID- 11868999 TI - Reduction of Kt/V by food intake during haemodialysis. AB - BACKGROUND: Urea kinetic modelling is a method for verifying that the amount of dialysis prescribed equals the amount of dialysis delivered. DOQI guidelines help to assess the causes of a low Kt/V: fistula integrity, treatment duration, methods of obtaining BUN samples, dialysis machine and patient specific variables. OBJECTIVE: To demonstrate the relation between food intake during HD and measurement of Kt/V. METHODS: Prospective study on 14 patients. We measured the amount of dialysis during the second week, with and without food intake during HD. HD prescription (blood flow, dialysate flow, dialyser, dry weight, heparin) was not changed. Statistical analysis was performed using Paired T test. CONCLUSIONS: 1. There was no significant change in protein catabolic rate, average urea concentration and intradialysis gain with food intake. 2. There was a significant change in levels of Kt/V using Gotch formula and Dugirdas formula with food intake. PMID- 11869000 TI - A personal reflection identifying how the use of a professional portfolio helped direct my career development. PMID- 11869001 TI - Epidemiology and management of hepatitis C in haemodialysis patients: an informal multidisciplinary review. PMID- 11869002 TI - The promise of tissue engineering. PMID- 11869005 TI - Incidental carcinoma of the gallbladder. AB - Incidental gallbladder carcinoma (GBC) is a difficult management issue as there are no established guidelines. Laparoscopic cholecystectomy is associated with increased dissemination of the tumour cells (both in the peritoneal cavity and port sites). Depth of tumour invasion (T stage) and positive surgical margins are the most important prognostic factors, although tumour differentiation, lymphatic, perineural and vascular invasion may also affect the outcome. Simple cholecystectomy is adequate for mucosal (T1a) lesions only. For T1b tumours port site/wound excision with second radical operation (probably extended cholecystectomy -- wedge liver excision with regional lymphadenectomy) should be advised. T2 tumours should be treated with second radical operation (extended cholecystectomy or excision of medial liver segments 4b and 5 or 4, 5 and 8 with regional lymphadenectomy with or without excision of the extra-hepatic bile duct). Few T3 tumours can be cured and in some survival time may be prolonged by a second radical operation. More extensive liver resection (segments 4b and 5 or segments 4, 5 and 8) with regional lymphadenectomy with excision of the extra hepatic bile duct should be advised. A second radical operation may palliate some T4 tumours. In the absence of extensive nodal disease, this operation may prolong the survival time. Excision of the extra-hepatic bile duct should be undertaken whenever the tumour involves the cystic duct margin or the extra-hepatic biliary tree. Epidemiology, risk factors, aetiopathogenesis and the modes of spread of GBC are discussed in relation to appropriateness of the second radical operation. Indications, types and role of the second radical operation are discussed. PMID- 11869006 TI - Optimizing surveillance mammography following breast conservation surgery. AB - AIMS: Prompt detection and treatment of local recurrence (LR) following breast conservation surgery (BCT) may improve subsequent survival. Following early reports demonstrating increased LR in the first years after surgery, a practice of surveillance mammography starting 1 year from diagnosis has become established. Increasing use of adjuvant chemotherapy with adriamycin-containing regimens has resulted in radiotherapy being postponed, so that the first mammogram coincides with the acute radiotherapy reaction, resulting in patient discomfort and poor quality films. We wished to determine if the 1-year mammogram could safely be omitted. METHODS: We reviewed 1151 consecutive patients treated with BCT for in situ, stage I or II disease over a 10-year period. All patients had clear resection margins and, where indicated, underwent axillary surgery and adjuvant treatment. This consisted of radiotherapy (40 Gy with a 5 Gy boost), chemotherapy and/or tamoxifen. RESULTS: Overall, the 5-year actuarial rate of local recurrence was 4.8%. The cumulative risk of LR at 12 months was 0.3% (three patients) and 1.9% (20 patients) at 24 months. This included all cases of high grade DCIS (>30 mm in size) and all but one tumour with a Nottingham Prognostic index (NPI) of >6.0 which recurred locally. CONCLUSION: Even including those patients generally accepted to be at high risk of LR, the cumulative risk of LR was only 0.3% at 12 months from surgery. We now therefore schedule routine biennial mammography from time of surgery except for those with high risk of early local recurrence such as extensive, high-grade in situ disease. PMID- 11869007 TI - Telomerase activity and Bcl-2 expression in human breast cancer. AB - AIMS: Telomerase is a ribonucleoprotein that synthesizes telomeres and plays an important role in cellular immortalization. Bcl-2 gene encodes for a mitochondrial protein thought to prevent apoptosis of normal cells. We previously reported telomerase activity in 74% of human invasive breast cancers and detected a significant association between telomerase activity and prognostic parameters such as nodal status, tumour size and cellular proliferation. We hypothesized that telomerase reactivation in human breast cancer was associated with increased immunohistochemical expression of Bcl-2. METHODS: Bcl-2 immunohistochemical expression was determined in 25 infiltrating breast carcinomas with known telomerase activity (17 telomerase-positive and 8 telomerase-negative). The percentage of strongly and moderately stained tumour cells for Bcl-2 was determined by a breast pathologist who was blinded to telomerase data. Fisher's exact test was used to examine the association between telomerase activity and Bcl-2 expression. RESULTS: The median percentage of strongly stained tumour cells was 50% for telomerase-positive tumours (range, 0--100%) and 45% for telomerase negative tumours (range, 0--100%). Twelve (70%) of 17 telomerase-positive tumours expressed strong or moderate Bcl-2 staining in >50% of tumour cells compared with six (75%) of eight telomerase-negative tumours (P=1.0). CONCLUSION: Telomerase reactivation seems to be independent of Bcl-2 protein expression in human breast cancer. PMID- 11869008 TI - Mucinous adenocarcinoma of the small bowel with peritoneal seeding. AB - AIMS: Small bowel adenocarcinoma has an extremely poor prognosis because of delayed diagnosis and the presence of advanced disease. Carcinomatosis associated with a small bowel primary cancer has not been reported to be successfully treated in the past. METHODS: The clinical information prospectively recorded on six patients with carcinomatosis from small bowel adenocarcinoma was reviewed. All of these patients were treated with an aggressive local--regional strategy that utilized cytoreductive surgery plus peri-operative intraperitoneal mitomycin C and 5-fluorouracil. RESULTS: Disease control in the abdomen and pelvis was achieved in four of these patients. Their median survival was 12 months with one patient alive and well at 4.5 years. CONCLUSIONS: Small bowel adenocarcinoma continues to provide a surgical challenge. Complete resection of all visible disease combined with intraperitoneal chemotherapy to eradicate microscopic residual disease should be considered as an option in patients with carcinomatosis. PMID- 11869009 TI - Measuring gelatinase activity in colorectal cancer. AB - AIMS: Gelatinases (MMP-2, -9) are the most extensively studied MMPs in cancer. The aim of the study was to determine the levels of active and latent forms of the gelatinases in paired colorectal tumour and normal tissue ( n=77) and correlate these with pathological stage. METHODS: Gelatinase levels were compared following the techniques of gelatin zymography (active and latent) and the novel gelatinase activity assays (total and endogenous/active). RESULTS: Both latent and active MMP-2 and MMP-9 lysis bands (zymography) and both total (active and latent) MMP-9 and endogenous (active) MMP-9 and MMP-2 levels (activity assays) were greater in tumour than normal colorectal tissue. Total MMP-2 and MMP-9 levels as determined by activity assays correlated with both the Dukes staging (e.g. total MMP-9 in tumours: adenoma, 1.0 (0.3--3.6); Dukes A, 9.6 (2.4--35.4); Dukes B, 14.7 (1.5--103.9); Dukes C, 22.3 (2.2--57.9) and Dukes D, 37.4 (2.1- 47.0) ng/mg protein) and with lymphatic invasion (e.g. total MMP-9; in tumours which had undergone lymphatic invasion, 22.7 (2.1--57.9) and those with no lymphatic invasion, 14.0 (1.5--103.9) ng/mg protein). CONCLUSIONS: Both gelatinases were upregulated in tumour tissue, however total and not endogenous active levels correlated with the pathological stage of the tumour. Therefore gelatinases may only be activated when required for tumour invasion and metastasis. PMID- 11869010 TI - Extracellular matrix proteins and chemoradiotherapy: alpha5beta1 integrin as a predictive marker in rectal cancer. AB - AIMS: To investigate the effects of extracellular matrix (ECM) protein expression on the rates of apoptosis and proliferation in rectal cancers and subsequent response to chemoradiotherapy (CRT). METHODS: The expression of fibronectin, collagen IV, laminin and the fibronectin receptor (FnR, alpha5beta1 integrin) were analysed in 32 pre-treatment rectal cancer biopsies by immunohistochemistry. ECM expression was correlated with tumour mitotic index (MI), apoptotic index (AI) and histopathological response to CRT. RESULTS: 18/32 cancers showed a poor response and 14/32 a good response (5/14 with complete pathological response) to CRT. Moderate to strong staining was seen in 22/32 cancers for fibronectin, 5/32 for collagen IV and 18/32 for laminin. Tumour FnR was related to stromal fibronectin content, and was significantly associated with CRT response; good responders having higher FnR expression compared to poor responders. No association was found between FnR expression and either MI or AI in pre-treatment biopsies, nor between MI or AI and CRT response. CONCLUSIONS: Tumour FnR expression is independent of MI and AI, and may serve as a useful marker for CRT response in rectal cancer. PMID- 11869011 TI - Chemoembolization for primary liver cancer. AB - AIMS: For most patients with primary liver cancer surgical treatment is not feasible and prognosis without treatment is poor. We aimed to assess the morbidity and efficacy of transarterial chemoembolization (TACE) with lipiodol and mitomycin C in these patients in a prospective case-control study. METHODS: From August 1996 to May 2000 22 patients with non-resectable hepatocellular carcinoma were treated with TACE. In case of radiological or tumour-marker response, treatment was repeated after 4--6 weeks, up to seven times per patient. RESULTS: Morbidity was 23% and usually minor, no patient died within 30 days of treatment. A decrease in size of the reference tumour or constant tumour-size in CT-scan were observed in 14 of 20 patients (70%) and of the 19 patients with elevated AFP-serum levels 12 (63%) had an AFP reduction following treatment. The median survival time was 14 months with a 1- and 2-year survival rate of 69% and 29%, respectively. Survival was not different in radiological or AFP responders vs non-responders. CONCLUSION: While TACE with lipiodol and mitomycin C for primary liver cancer is associated with considerable antitumoural efficacy, as demonstrated by tumour marker and radiological response, an effect on patient survival is not evident. New treatment options with an impact on survival are needed for these patients. PMID- 11869012 TI - Primary squamous cell carcinoma of the thyroid. AB - AIMS: To investigate the clinical features and treatment protocol of squamous cell carcinoma of the thyroid (SCCT). METHOD: Clinical records of four SCCT patients treated during 1985--99 were retrospectively reviewed and evaluated. RESULTS: Two patients who underwent surgical excision plus radiotherapy died of local tumour recurrence, 6 and 13 months, respectively, post-operatively. One who accepted surgery only died 4 months later of respiratory distress. The fourth patient who had radical surgery coupled with radiotherapy and chemotherapy was disease-free at 26-month follow-up. CONCLUSION: SCCT is a very rare but highly malignant carcinoma. Early diagnosis and an aggressive radical surgery portend a better prognosis. Radiotherapy and chemotherapy are secondary applicable methods for the patients with certain condition. PMID- 11869013 TI - Endoscopic-assisted intrathoracic oesophagogastrostomy without thoracotomy for tumours of the lower oesophagus and cardia. AB - AIMS: This study aimed to evaluate the efficacy of a novel technique enabling a trans-hiatal oesophagectomy with intrathoracic anastomosis under direct vision, without thoracotomy. METHODS: Trans-hiatal dissection of the oesophagus was performed using direct and laparoscopic visualization. The oesophagus was transected above the tumour with a linear endo-GIA-2 60 mum stapler. The stomach was transected and a gastric tube fashioned. The anvil of an appropriately sized CEEA circular stapler was modified enabling it to flatten. It was attached to a novel delivery system introduced under direct vision along a guidewire into the stapled oesophagus. The anvil was realigned to its original position in the distal oesophagus, docked with the body of the stapler and an intrathoracic anastomosis performed. RESULTS: Ten patients (female n=3, male n=7) aged from 39- 77 years (mean age 65 years), ASA 2--3 with distal third tumours were treated. Duration of procedure ranged from 2--5 hours (mean 4 hours). One patient suffered a post-operative chest infection and an anastomotic leak treated successfully with a self-expanding metal stent. Hospital stay ranged from 6--28 days (mean 17 days). There was no mortality. CONCLUSION: This technique allows a safe intrathoracic anastomosis to be performed trans-hiatally under direct vision, avoiding the need for thoracotomy in patients with high comorbidity. PMID- 11869014 TI - Surgical treatment of pulmonary metastases: experience with 40 patients. AB - AIMS: Combinations of surgery and chemotherapy have a favourable impact on survival in the treatment of disseminated neoplastic disease isolated to the lung. Sample and technical factors have made the reproduction of the published results difficult. METHODS: In this study we report experience over 10 years. RESULTS: From 1989 to 1999 40 patients underwent metastasectomy. Thirty received chemotherapy. The median survival is 51 months, similar to other published series. CONCLUSION: Survival benefit can be observed in small series of such cases. PMID- 11869015 TI - Value of positron emission tomography for lung cancer staging. AB - OBJECTIVE: The therapeutic strategy in non-small-cell lung cancer (NSCLC) requires exact staging of tumour invasion (T) as well as differentiation between ipsi- and contralateral lymph node invasion (N1/2 vs N3). [18F]FDG-positron emission tomography (FDG-PET) has been shown to detect invaded N with high accuracy while correct determination of T appears to be unclear. The purpose of this prospective study was to evaluate benefit and necessity of 18FDG-PET as an additive to conventional staging modalities. METHODS: Forty patients with suspected non-small-cell lung cancer (NSCLC) were staged by means of computed tomography (CT), bronchoscopy, mediastinoscopy and bone scintigraphy. Additionally, attenuation corrected FDG-PET of the thorax was performed pre operatively for analysis of T and N topography. After surgical resection with radical lymphadenectomy T and N staging results of CT and PET were compared with the pathological diagnoses. Specificity, sensitivity, positive predictive value and accuracy of CT and PET were calculated. RESULTS: Twenty three squamous cell carcinomas, 14 adenocarcinomas, and three non-malignant tumours were found. Accuracy of CT-T was 0.75 and of PET-T 0.78; accuracy of CT-N was 0.78 and of PET N 0.80. By combination of CT-T and PET-T accuracy was 0.88. Combination of CT-N and PET-N yielded an accuracy of 0.90. In two out of three cases, PET correctly determined T0. In two cases non-malignant inflammatory lymph nodes were falsely staged as malignant by PET. CONCLUSIONS: Adequate pre-operative T- and N-staging is possible with both CT and FDG-PET. Accuracy can be improved by combination of CT and FDG-PET. FDG-PET is superior to CT in order to differentiate between malignant and benign tumours. However, acute inflammation can mimic malignant lymph node invasion. FDG-PET is justified as a supporting staging measure in cases presenting unclear differentiation between N2 and N3 after conventional staging and is helpful in cases with unclear cell type of the primary tumour. PMID- 11869016 TI - Focal neuroendocrine differentiation in non-small cell carcinoma is not predicted by pre-operative routine investigation. AB - AIMS: Between 10 and 35% of resected non-small cell carcinomas of the lung show focal neuroendocrine differentiation. The significance of this is uncertain but recent studies have suggested that these tumours may be more responsive to chemotherapeutic agents than other non-small cell carcinomas. This study was undertaken to determine if focal neuroendocrine differentiation in surgically resected non-small cell carcinomas could be predicted in patients who had undergone traditional routine pre-operative diagnostic investigations. METHODS: Sixty-six consecutive resections for non-small cell carcinoma were reviewed for the presence of focal neuroendocrine morphology or positive immunohistochemical staining with neuroendocrine markers. Pre-operative biopsy and cytology samples from these patients were then sought for similar assessment. RESULTS: Ten of the 66 cases showed either morphological or immunohistochemical evidence of neuroendocrine differentiation. All 10 patients had undergone pre-operative investigation including bronchoscopy but in only four were pre-operative diagnostic samples obtained (two cytology and two biopsies). None of these specimens showed features to suggest the presence of neuroendocrine differentiation within the resected tumours. CONCLUSION: The presence of focal neuroendocrine features in an unselected series of resected non-small cell carcinomas was not predicted by traditional pre-operative diagnostic investigations This indicates that focal neuroendocrine differentiation in patients with unresectable non-small carcinoma cannot be reliably identified for inclusion in clinical trials using traditional investigate approaches. PMID- 11869017 TI - Prognostic value of proliferating cell nuclear antigen in Wilms' tumour in children. AB - AIMS: To evaluate the prognostic value of index Proliferating Cell Nuclear Antigen (PCNA) in Wilms' tumour in children. METHODS: The study comprised 64 children aged from 2 days to 13 years treated according to the SIOP (Society International of Oncology Paediatric) and accepted by the PPGGL (Polish Paediatric Group for the Treatment of Solid Tumours). The studies were conducted on tumour tissue removed during surgery, fixed in formalin and embedded in paraffin blocks. Sections (4 microns) were evaluated by immunohistochemistry, using the peroxidase method to determine the expression of PCNA in Wilms' tumour cells by primary monoclonal antibody NCL-PCNA from Novocastra. RESULTS: The percentage of immunopositive cells in particular fragments of the tumour ranged from 0--93%, mean 30.5%, median 25.5%. Mean and median values enabled division of children into two groups: Group A, where the percentage of cells staining with anti-PCNA was <30% and Group B, where this percentage was >30%. The expression of PCNA was evaluated in various stages of advancement, various histological types and depending on the course of disease. The studies revealed the correlation between index PCNA and stage of advancement P<0.01, index PCNA and histological type of Wilms' tumour P<0.025. Moreover we observed that deaths were found more frequently in tumours with index PCNA >30%, P<0.001. CONCLUSIONS: PCNA is a useful prognostic factor in Wilms' tumour in children. PMID- 11869018 TI - Lymphatic mapping and sentinel lymph node biopsy in squamous cell carcinoma of the lower lip. AB - AIMS: The risk of metastases to the submandibular and submental lymph nodes in squamous cell carcinoma (SCC) of the lower lip is closely related to the primary tumour size and the differentiation of the tumour. In order to determine the feasibility of the technique and the possible metastatic lymph nodes in SCC of the lower lip, intraoperative lymphatic mapping and sentinel lymph node biopsy was performed in patients with tumour size greater than 2 cm (T2) and clinically non-palpable regional lymph nodes (N0). METHODS: Intraoperative lymphatic mapping with patent blue dye was performed in 20 patients with SCC of the lower lip. The stained lymph node (sentinel) was identified in each patient and sent for frozen section analysis in order to verify tumour metastasis. All patients had undergone bilateral suprahyoid neck dissection at the same stage. RESULTS: Three of the patients were female and 17 were male. The median age was 66. Sentinel lymph nodes were identified in 18 of the patients (90%). Intraoperative or post operative histopathologic examination of the sentinel lymph node showed tumour metastasis in three of the patients (16.6%). The histopathologic examination of the remaining 15 patients whose sentinel lymph nodes were free of metastasis, showed no metastasis in the non-sentinel lymph nodes. In two of the three patients with metastatic sentinel lymph nodes, non-sentinel lymph nodes were free of metastases. There were no false negative results and no local or systemic complications of the technique were seen among the patients. CONCLUSIONS: Intraoperative lymphatic mapping and sentinel lymph node biopsy is feasible in patients with SCC of the lower lip who have large tumour size and non-palpable regional lymph nodes. The technique may help to avoid neck dissection when the patient has negative sentinel lymph node and when positive provides useful information for more effective radical treatment. PMID- 11869020 TI - Cytoreduction and intraperitoneal chemotherapy for the management of peritoneal carcinomatosis, sarcomatosis and mesothelioma. AB - Despite new developments in multi-modality treatments, complete resection remains as an absolute requirement for cure of gastrointestinal cancer. We have reported benefits from combined treatment with complete cytoreduction and intraperitoneal chemotherapy. This has been achieved with low morbidity and mortality. Success in the surgical management of peritoneal surface malignancy depends on the surgeon's ability to complete complex cytoreductive procedures so that only microscopic residual disease remains. This paper describes the current strategy that the surgical oncologist should pursue in the treatment of patients with peritoneal carcinomatosis, sarcomatosis and mesothelioma. Technical details required for this surgery include patient position, incision and exposure, complete lysis of adhesion, electroevaporative dissection with irrigation and suction to preserve the translucent quality of tissues, peritonectomy procedures, proper positioning of tubes and drains for intraperitoneal chemotherapy, and reconstructive surgery. Understanding the treatment and mastery of surgical skills to manage the peritoneal surface spread of cancer has led to long-term survival of selected patients. Combination of this treatment strategy with proper patient selection has reduced the mortality and morbidity. The success of cytoreductive surgery and perioperative intraperitoneal chemotherapy depends on a long-term dedication to achieve the full potential of a curative outcome. Our unit has continued to achieve good results over two decades as improved results of treatment have evolved. PMID- 11869019 TI - Wound complications following pre-operative radiotherapy for soft tissue sarcoma. AB - AIMS: We analysed wound complications in 43 patients with soft tissue sarcoma who were treated with combined pre-operative radiotherapy and surgery. METHODS: All patients received the same protocol of pre-operative radiotherapy at our institution. RESULTS: Thirty-six (84%) patients developed acute skin toxicity following radiotherapy. After wide local excision, 15 patients required primary soft tissue reconstruction with vascularized muscle transfer and four patients underwent free skin flap to enable wound closure as part of their primary surgery. Nineteen patients (44%) developed post-operative wound complications including 10 (23%) patients who required an additional surgical procedure. Four (27%) patients developed flap necrosis in a group of 15 who underwent primary vascularized soft tissue transfer. All required a second vascularized muscular flap. One elderly patient, who had grade 3 acute radiation skin toxicity, had an arterial graft and total hip arthroplasty for a femoral artery aneurysm and an avascular necrosis of the hip, respectively. In our series, age (> or = 40 years) was the only impact factor influencing wound complication after surgery following radiotherapy (P=0.06). CONCLUSIONS: Site of tumour, radiation field size, surgical resection volume, grade of acute radiation toxicity, co-morbidity, and smoking were not demonstrated to have predictive value in wound complication following pre-operative radiotherapy. Although previous papers suggested that vascularized soft tissue transfer could be useful reducing wound morbidity, our results could not confirm this. PMID- 11869021 TI - Ancient schwannoma masquerading as a thyroid mass. AB - Schwannomas are benign, encapsulated nerve sheath cell neoplasms. Cervical sympathetic chain (CSC) schwannomas are rare, with less than 50 cited cases in the literature. CSC schwannomas may mimic a number of parapharyngeal masses. We report a rare variant, "ancient" schwannoma, which presented cytologically and radiologically as a thyroid mass. This is the first report of a CSC schwannoma mimicking a thyroid mass and the first report of an ancient schwannoma of the CSC. PMID- 11869022 TI - Primary endodermal sinus (yolk sac) tumour of the liver. AB - Primary endodermal sinus tumour of the liver is an extremely rare neoplasm. A 37 year-old man presented with a mass in the left lobe of the liver and a raised serum alpha-fetoprotein concentration. A left hepatic lobectomy was performed. Histological examination revealed an endodermal sinus tumour of the liver. There was no evidence of an extrahepatic primary source. PMID- 11869023 TI - Breast abscess as the initial presentation of squamous cell carcinoma of the breast. AB - Breast abscesses are common and rarely harbour an underlying malignancy. Antibiotics and drainage by aspiration are the mainstays of treatment. We encountered two post-menopausal patients aged 48 and 70 who presented with acute breast abscesses that had an underlying squamous cell carcinoma of the breast. Infective aetiology was considered in each case and pre-operative diagnosis is possible but requires a high level of suspicion to avoid a delay in diagnosis. Aspirate from the abscess should be sent for both culture and cytology and if drainage is done, then biopsy of the abscess cavity is essential. Clinicians should be aware of the association of this rare primary neoplasm of the breast. PMID- 11869024 TI - Metastatic adenocarcinoma arising from a small bowel duplication cyst. AB - A case report of adenocarcinoma arising from a small bowel mesenteric cyst is presented. A discussion and review of the relevant literature then follows. PMID- 11869025 TI - Hyperthermia in anticancer treatment. PMID- 11869028 TI - NPY Y1 receptors are present in axonal processes of DRG neurons. AB - Using a sensitive immunohistochemical method, the localization of the neuropeptide Y (NPY) Y1 receptor (Y1R) was studied in contralateral and ipsilateral dorsal root ganglion (DRG) neurons of rats subjected to different unilateral manipulations with focus on their axonal processes and projection areas. Y1R-like immunoreactivity (LI) was observed in the contralateral sciatic nerve and dorsal roots of lesioned rats, and double staining revealed colocalization with calcitonin gene-related peptide (CGRP). Y1R-LI was also seen in fibers close to and even within the epidermis. A fairly small number of nerve endings double-labeled for Y1R and CGRP were present in the dorsal horn. After unilateral crush of the sciatic nerve Y1R- and CGRP-LI accumulated in the same axons proximal to the lesion. After dorsal rhizotomy CGRP-LI was strongly reduced in the ipsilateral dorsal horn. No certain change was observed for Y1R- or NPY LI, but Y1R/CGRP double-labeled nerve endings disappeared after the lesion. These results strongly suggest centrifugal transport of Y1Rs in DRG neurons, mainly to the peripheral sensory branches. To what extent these Y1Rs are functional has not been analyzed here, but a recent study on Y1R null mice provides evidence for involvement of prejunctional Y1Rs in peripheral sensory functions PMID- 11869029 TI - Human bone marrow stem cells exhibit neural phenotypes and ameliorate neurological deficits after grafting into the ischemic brain of rats. AB - There is now evidence to suggest that bone marrow mesenchymal stem cells (MSCs) not only differentiate into mesodermal cells, but can also adopt the fate of endodermal and ectodermal cell types. In this study, we addressed the hypotheses that human MSCs can differentiate into neural cells when implanted in the brain and restore sensorimotor function after experimental stroke. Purified human MSCs were grafted into the cortex surrounding the area of infarction 1 week after cortical brain ischemia in rats. Two and 6 weeks after transplantation animals were assessed for sensorimotor function and then sacrificed for histological examination. Ischemic rats that received human MSCs exhibited significantly improved functional performance in limb placement test. Histological analyses revealed that transplanted human MSCs expressed markers for astrocytes (GFAP(+)), oligodendroglia (GalC(+)), and neurons (beta III(+), NF160(+), NF200(+), hNSE(+), and hNF70(+)). The morphological features of the grafted cells, however, were spherical in nature with few processes. Therefore, it is unlikely that the functional recovery observed by the ischemic rats with human MSC grafts was mediated by the integration of new "neuronal" cells into the circuitry of the host brain. The observed functional improvement might have been mediated by proteins secreted by transplanted hMSCs, which could have upregulated host brain plasticity in response to experimental stroke. PMID- 11869030 TI - Striatal AMPA receptor binding is unaltered in the MPTP-lesioned macaque model of Parkinson's disease and dyskinesia. AB - Long-term levodopa or dopamine agonist treatment in the MPTP-lesioned primate model of Parkinson's disease elicits dyskinesia, which is phenotypically similar to levodopa-induced dyskinesia in patients with Parkinson's disease. AMPA receptor antagonists have previously been shown to have both anti-parkinsonian and anti-dyskinetic actions in MPTP-lesioned primates, suggesting that AMPA receptor transmission is functionally overactive under these conditions. In this study, we investigated the level of striatal AMPA receptor binding in the MPTP lesioned primate using the selective AMPA ligand (3)H-(S)-5-fluorowillardiine. AMPA receptor binding was studied in non-parkinsonian, non-dyskinetic parkinsonian, and dyskinetic macaques. Striatal AMPA receptor binding was not different in any of the treatment groups (P > 0.05). Although AMPA receptor mediated transmission is functionally overactive in Parkinson's disease and dyskinesia, changes in striatal AMPA receptor levels are not likely to be the cause of such movement disorders. PMID- 11869031 TI - Perinatal asphyxia induced neuronal loss by apoptosis in the neonatal rat striatum: a combined TUNEL and stereological study. AB - Perinatal asphyxia can lead to cell damage in various regions of the brain, such as the neostriatum. In this study, we investigated the mechanism of cell death that leads to neuron loss in the neostriatum of rat pups. Asphyxia was induced by immersing fetus-containing uterus horns in a water bath at 37 degrees C for 20 min. This led to an increase in mortality rate (+/- 40%) compared to control pups (0%). TUNEL-positive cell profiles were visible in all groups at postnatal day (P) 2, P8, and P15, peaking at P8. A significant increase of 40% at P8 and 45% at P15 in the number of TUNEL-positive cell profiles was observed in asphyctic rats compared to control rats. Nuclear condensation and fragmentation was visible with the DNA stain Hoechst 33342. Furthermore, laser-scanning confocal microscopy showed multiple DNA fragments in TUNEL-positive cell profiles. We found a decrease of 16% in the total number of striatal neurons in the asphyctic pups compared to the control pups at 21 days postasphyxia using stereology. These data show that asphyxia causes exaggerated apoptotic cell death during the first week of life and as a consequence a small amount of neuron loss in the neostriatum. PMID- 11869032 TI - Survival signaling and selective neuroprotection through glutamatergic transmission. AB - In the brain, alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA) receptors mediate glutamatergic neurotransmission and, when intensely activated, can induce excitotoxic cell death. In addition to their ionotropic properties, however, AMPA receptors have been functionally coupled to a variety of signal transduction events involving Src-family kinases, G-proteins, and the mitogen activated protein kinase (MAPK). In the present study, we tested whether AMPA receptors are linked to appropriate signaling events in order to prevent neuronal injury and/or enhance recovery. AMPA stimulation in hippocampal slice cultures caused the selective activation of MAPK through the upstream activator MAPK kinase (MEK). Inhibition of either component of the AMPA receptor--MAPK pathway potentiated cellular damage due to serum deprivation, suggesting that this pathway facilitates compensatory signals in response to injury. Correspondingly, positive modulation of AMPA receptors with the Ampakine 1-(quinoxalin-6 ylcarbonyl)piperidine (CX516) enhanced MAPK activation and reduced the extent of synaptic and neuronal degeneration resulting from excitotoxic episodes. CX516 was neuroprotective when infused into slices either before or after the insult. The Ampakine derivative also elicited neuroprotection in an in vivo model of excitotoxicity as evidenced by reduction in lesion size and preservation of two different types of neurons. Interestingly, the AMPA receptor--MAPK pathway selectively protects against excitotoxicity since enhancing the pathway did not protect against the nonexcitotoxic, slow pathology initiated by lysosomal dysfunction. The results indicate that glutamatergic communication is important for cellular maintenance and that AMPA receptors activate survival signals to counterpoise their own excitotoxic potential. PMID- 11869033 TI - Stable expression of the alkaline phosphatase marker gene by neural cells in culture and after transplantation into the CNS using cells derived from a transgenic rat. AB - Multipotent stem cells and more developmentally restricted precursors have previously been isolated from the developing nervous system and their properties analyzed by culture assays in vitro and by transplantation in vivo. However, the variety of labeling techniques that have been used to identify grafted cells in vivo have been unsatisfactory. In this article we describe the characteristics of cells isolated from a transgenic rat in which the marker gene human placental alkaline phosphatase (hPAP) is linked to the ubiquitously active R26 gene promoter. We show that hPAP is readily detected in embryonic neuroepithelial stem cells, neuronal-restricted precursor cells, and glial-restricted precursor cells. Transgene expression is robust and can be detected by both immunocytochemistry and histochemistry. Furthermore, the levels of hPAP on the cell surface are sufficient for live cell labeling and fluorescence-activated cell sorting. Expression of hPAP is stable in isolated cells in culture and in cells transplanted into the spinal cord for at least 1 month. We submit that cells isolated from this transgenic rat will be valuable for studies of neural development and regeneration. PMID- 11869034 TI - Selective and AMPA receptor-dependent astrocyte death following prolonged blockade of glutamate reuptake in rat cerebellar cultures. AB - In this study we examined the effects of prolonged l-trans-pyrrolidine-2,4 dicarboxylate (PDC)-induced glutamate reuptake blockade on the viability of glial cells in cerebellar granule cell cultures. Immunofluorescence staining for the glial-specific intermediate filament protein, GFAP, revealed that the PDC- induced increase of extracellular glutamate concentration was accompanied by increased astrocyte death, while neurons and oligodendrocytes remained intact and viable. Astrocytic cell death was manifested as fragmentation of processes and cell bodies. The selective astrocyte death was completely prevented by the competitive alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA)/kainate receptor antagonist, NBQX (10 microM), whereas MK-801 (10 microM), a noncompetitive blocker of N-methyl-D-aspartate receptors, gave only partial protection. Double staining for GFAP and the AMPA receptor subunits GluR2/3 showed that astrocytes had much higher immunoreactivity for GluR2/3 than neurons or oligodendrocytes, suggesting that the number of AMPA receptors is likely to be higher on astrocytes. Furthermore, we employed real-time RT-PCR to measure GluR1 4 subunit mRNA expression in control and PDC-exposed cultures. Following treatment with PDC, GluR1 and GluR4 mRNAs were reduced by 40% and GluR3 was reduced by 70% relative to control levels. In contrast, GluR2 expression was not affected by the PDC treatment, indicating that GluR3 was the dominant type of AMPA receptor subunit expressed on astrocytes. Our results show that astrocytes appear to be more vulnerable than neurons or oligodendrocytes to a gradual increase in the extracellular glutamate concentration, suggesting that astrocytes may be critically involved in the pathophysiology of slowly developing chronic neurodegenerative disorders. PMID- 11869036 TI - Complement activated C4d immunoreactive oligodendrocytes delineate small cortical plaques in multiple sclerosis. AB - C4d-immunoreactive complement-activated oligodendrocytes (C4d-CAOs) have been described in several neurodegenerative diseases but have not been studied in multiple sclerosis (MS). Here we report that such CAOs delineate miniature MS plaques of 300-500 mum diameter. They are devoid of myelin and are surrounded by a rim of activated microglia intermingled with the C4d-CAOs. Although C4d immunostained periaxonal oligodendroglial processes are often swollen, the axons of passage appear undamaged and extend through the demyelinated plaque area. No immunostaining with other components of the complement cascade (C1q-C9) was observed in association with these miniature plaques. However, in large MS lesions, C1q-C9 immunoreactive fibers were present, indicating complete activation of the complement cascade in these more developed lesions. It is possible that the miniature plaques, bordered by C4d-CAOs, represent the earliest stage of plaque development, preceding even the larger, transient plaques frequently observed in serial MRI studies. The association of CAOs with miniature areas of demyelination suggests a direct attack on oligodendroglial cells by the early complement components as an initiating event in MS. Incomplete complement activation indicates that this step may be reversible, whereas full and persistent activation as observed in large MS lesions may lead to death of oligodendroglia with permanent axonal damage. PMID- 11869035 TI - Biotinylated dextran amine as a marker for fetal hypothalamic homografts and their efferents. AB - We have explored the use of biotinylated dextran amine (BDA) as a marker for labeling fetal brain grafts and their connections with the host. As a model system we used transplantation of the hamster suprachiasmatic nucleus, the site of an endogenous biological clock governing circadian rhythms. Similar transplants into arrhythmic hosts have been shown to restore behavioral function with a period specific to the donor. For locomotor rhythms, efferent connections are not necessary. For other responses, including endocrine rhythms, efferent connections may be necessary. In order to visualize homografts and their efferents, injections of BDA, an anterograde tracer, were made into the anterior hypothalamic (AH) region containing the SCN or into the dorsal cortex (CTX) of fetal hamster brains. The fetal AH or CTX was microdissected out and stereotaxically implanted into the third ventricle of intact, adult hamsters. After 2, 4, 8, or 12 weeks, hosts were sacrificed and their brains were processed for detection of BDA by either histochemistry or immunofluorescence. BDA intensely labeled graft neurons, their dendrites, and axons with minimal or no spread to the adjacent host brain. Labeled graft axons could be followed for long distances (>1 mm) into the host brain and graft-derived varicosities formed close contacts with host neurons. BDA-labeled graft neurons, located at the perimeter of the graft, also extended dendrite-like processes into the host parenchyma. We conclude that BDA is a useful marker for fetal homografts and their efferents for survival times of less than 2 months. PMID- 11869037 TI - Nestin expression in ganglioglioma. AB - It has been suggested that gangliogliomas represent a neoplastic transformation of a dysplastic focus or heterotopia. Other theories propose that gangliogliomas arise from multipotent stem cells with the ability to differentiate along glial and neuronal cell lines. Our goal was to characterize the expression of nestin, a neuroepithelial precursor/stem cell antigen, in gangliogliomas along with other pathological and clinical features of this entity. The clinical and operative features of 18 recent cases meeting the histological criteria for ganglioglioma were reviewed. The expression of nestin, microtubule-associated protein 2 (MAP2), neurofilament, and glial fibrillary acidic protein (GFAP) was assessed by immunohistochemistry and confocal scanning laser microscopy. Abundant MAP2- and nestin-positive neuronal cells were found by immunohistochemistry in all 18 gangliogliomas. GFAP staining was found in reactive and lesional astrocytes but not in cells of neuronal morphology. Confocal microscopy demonstrated colocalization of nestin and MAP2 in select neuronal cells. The true lineage of gangliogliomas remains controversial. Our findings confirm the presence of cells within these lesions that harbor a persistent stem cell cytoskeletal protein (nestin). Further insight into the cytoskeletal derangement of nestin-positive neuronal cells may shed further light on the pathogenesis of gangliogliomas and its associated epilepsy. PMID- 11869038 TI - Facilitation of dopamine-mediated locomotor activity in adult rats following cholinergic denervation. AB - The dopamine hypothesis of schizophrenia postulates hyperactivity of dopaminergic neurotransmission in the mesolimbic system. However, the possible underlying causes for this dopaminergic overfunction are not well understood. Therefore, the main aim of this study was to examine the effect of central cholinergic denervation on dopamine-mediated functions. We also examined the effect of neonatal cholinergic denervation upon adult brain function. The immunotoxin 192 IgG-saporin causes severe lesions of the basal forebrain cholinergic system when infused into the lateral ventricles by targeting neurons expressing the p75 neurotrophin receptor. The toxin may also damage p75-expressing Purkinje neurons in the cerebellum. We have compared the behavioral effects of intracerebroventricular injections of 192 IgG-saporin to adult rats with that of injections to neonate rats. As expected, adult treated rats displayed an almost complete cholinergic denervation of forebrain corticohippocampal areas concomitant with a marked impairment in the Morris water maze. When tested as adults, neonatally treated animals had a less complete cholinergic denervation and showed lesser impairments in water maze behaviors. Interestingly, adult treated rats showed increased spontaneous horizontal activity and a remarkable increase in locomotor response to d-amphetamine as evidenced by increased horizontal and vertical activity. There were no marked changes of spontaneous or drug-induced locomotor activity in adult rats treated with 192 IgG-saporin as neonates. These results suggest that cholinergic denervation of the forebrain causes a marked enhancement of behavioral responses related to dopaminergic activity, probably mainly mediated presynaptically. However, it cannot be fully excluded that damage to noncholinergic systems, e.g., Purkinje cells, might contribute to the effects. The striking overreaction to dopaminergic stimuli, presumably caused by the cholinergic deficit, is discussed in relation to the suggested role of cholinergic malfunctioning in schizophrenia. PMID- 11869039 TI - Expression of dystroglycan and the laminin-alpha 2 chain in the rat peripheral nerve during development. AB - In Schwann cells, the transmembrane glycoprotein beta-dystroglycan comprises the dystroglycan complex, together with the extracellular glycoprotein alpha dystroglycan, which binds laminin-2 (alpha 2/beta 1/gamma 1), a major component of the Schwann cell basal lamina. To provide clues to the biological functions of the interaction of the dystroglycan complex with laminin-2 in peripheral nerves, we investigated the expression of beta-dystroglycan and the laminin-alpha 2 chain in rat sciatic nerve during development by immunoblot, immunofluorescence, and immunoelectron microscopic studies. The expression of beta-dystroglycan and the laminin-alpha 2 chain in the rat sciatic nerve was low and not confined to the Schwann cell outer membrane from embryonic day 18 to birth, when there was only an immature basal lamina assembly and no compact myelin formation by Schwann cells. However, the expression of these proteins increased markedly and became clearly localized to the Schwann cell outer membrane between birth and postnatal day 7, when both basal lamina assembly and compact myelin formation by Schwann cells progressed rapidly. From postnatal day 7 to adult, there was no remarkable change in the expression of these proteins. Our results support the hypothesis that the dystroglycan complex functions as an adhesion apparatus, binding the Schwann cell outer membrane with the basal lamina, and suggest that the dystroglycan complex plays a role in Schwann cell myelination through its interaction with laminin-2. PMID- 11869040 TI - Expression of herpes simplex virus type 2 protein ICP10 PK rescues neurons from apoptosis due to serum deprivation or genetic defects. AB - Previous studies have shown that the herpes simplex virus type 2 protein kinase ICP10 PK activates the Ras/MEK/MAPK pathway in nonneuronal cells. Here we report that ectopically expressed ICP10 PK has anti-apoptotic activity in various paradigms of neuronal cell death. Neuronally differentiated PC12 cells and primary murine hippocampal cultures transfected with an expression vector for ICP10 PK were protected from cell death resulting from growth factor withdrawal. Protection from apoptosis was also seen in ICP10 PK-transfected hippocampal neurons from the trisomy 16 mouse, a naturally occurring genetic abnormality the human analog of which is Down syndrome. Cells transfected with an expression vector for a mutant that lacks kinase activity were not protected, although it was expressed as well as ICP10 PK. The data indicate that ICP10 PK has a broad anti-apoptotic activity in neuronal cells which depends on a functional PK. PMID- 11869041 TI - The role of tissue transglutaminase (transglutaminase type II) for the intestinal manifestations of murine semi-allogenic graft-versus-host disease. AB - The intestinal manifestation of acute murine semi-allogenic graft-versus-host (GvH) disease is characterized by the occurrence of lymphocytic infiltrates in the lamina propria, by crypt hyperplasia and villous atrophy. In a histological respect, this animal model resembles human celiac disease. Tissue transglutaminase (tTG) (transglutaminase type II) has been identified to be the major B cell autoantigen in celiac disease. Furthermore, tissue transglutaminase has been implicated to be involved in its pathogenesis. Therefore, we aimed to investigate whether tissue transglutaminase is expressed in the intestines of GvH animals and whether its inhibition has any effect on the intestinal histology. Sera of patients with celiac disease and anti-tTG antibodies were purified. These antibodies were used for immuno-histochemistry of jejunal cryosection from GvH and syngenic control animals at day 6 after lymphocyte transfer. Furthermore, GvH mice were treated with antitTG antibodies and with the inhibitor of tissue transglutaminase monodansyl-cadaverine. The effect of this treatment on the development of crypt hyperplasia and villous atrophy were examined by light microscopy of hematoxylin-eosin (H&E) stained jejunal paraffin sections. We found a strong subepithelial expression of tissue transglutaminase in GvH animals but not in syngenic control mice. The localization of tTG seemed to be associated with the extracellular matrix (ECM). However, neither the treatment of GvH animals with anti-tTG antibodies nor the application of mono-dansyl-cadaverine prevented the development of crypt hyperplasia and villous atrophy. Similar to the situation in human celiac disease tissue, transglutaminase is highly expressed in the intestine of animals undergoing a semi-allogenic graft-versus host reaction. However, this enzyme is probably not involved in the development of crypt hyperplasia and villous atrophy in this animal model. PMID- 11869042 TI - Anti-CD44 monoclonal antibody (IM7) induces murine systemic shock mediated by platelet activating factor. AB - The cell adhesion molecule CD44 plays an important role in progression of autoimmune diseases or cancer. Administration of anti-CD44 monoclonal antibodies (mAbs) have been reported to have anti-inflammatory or anti-cancer activity. However, our evidence shows that intravenous administration of the anti-CD44 IgG2b mAb IM7 induces systemic shock in mice. To examine the character of systemic shock, the cutaneous excess vascular permeability was evaluated. Administered mAb markedly increased vascular permeability but its F(ab')(2) fragments did not induce a reaction. The platelet-activating factor (PAF) specific antagonist Y-24180 was effective in preventing IM7-induced extravasation, whereas anti-histaminergic and anti-serotonergic agents were not. Y-24180 also ameliorated hematocrit elevation and hypotension in mice treated with IM7. These results indicate that IM7-induced systemic shock is mediated by PAF. Because IM7 also binds human CD44, anti-CD44 immunotherapy using IM7 may be applied to the clinical treatment of autoimmune diseases or cancer. This study describes potential triggering pathways for shock that must be avoided through modification of the immunotherapy. PMID- 11869043 TI - Selective increase of autoimmune epitope expression on aged erythrocytes in mice: implications in anti-erythrocyte autoimmune responses. AB - We investigated the impact of changes occurring during red blood cell (RBC) ageing on the RBC-binding activity of pathogenic anti-erythrocyte monoclonal antibodies derived from autoimmune-prone New Zealand black (NZB) mice. As assessed by flow cytometric analysis on in vivo biotinylated RBCs, all five NZB derived anti-RBC mAb exhibited more efficient binding to aged RBCs than to young RBCs, and resulted in a selective elimination of more aged RBCs from the circulating blood. In addition, treatment of RBCs with proteases markedly enhanced the binding of all five anti-RBC mAb, raising the possibility that increased exposure of autoimmune epitopes on aged RBCs may be in part, a result of contacts with proteolytic enzymes during the lifetime of circulating RBCs. In marked contrast, the binding activity of mAb raised in non-autoimmune animals against antigens expressed on RBCs, such as CD44, CD47, CD147 and TER-119, was either decreased or unchanged with RBC ageing, and these epitopes, except for that recognized by anti-CD47 mAb, were highly sensitive to mild treatment with proteases. Our data unravel the unique molecular feature of RBC epitopes involved in autoimmune haemolytic anaemia, suggesting that membrane alterations in aged RBCs might play a significant role in the development of the autoantibody response to RBCs. PMID- 11869045 TI - Susceptibility to cyclosporin A-induced autoimmunity: strain differences in relation to autoregulatory T cells. AB - Cyclosporin A-induced autoimmunity (CsA-AI), also called autoimmune syngeneic graft-vs-host disease, is a thymus dependent, T cell mediated rodent animal model of disease and is considered to be an experimental model for human scleroderma. Since adoptive transfer of CsA-AI by effector T cells can be prevented by autoregulatory T cells, there may also be a role for dominant tolerance in the resistance of certain rat strains to develop clinical manifest CsA-AI. LEW rats have been reported to be susceptible, whereas BN rats are resistant to CsA-AI. In the present study we first demonstrate that PVG, but not DA rats, are susceptible to CsA-AI and that disease characteristics in PVG rats are comparable to LEW rats in terms of pathogenesis and T cell kinetics, although of more rapid onset and greater severity. Next, we examined whether the relative presence of autoregulatory T-helper cells, i.e. CD25+ and/or CD45RClow CD4 T cells, is increased in resistant BN and DA rats. The results obtained reveal that the genetically determined CD45RChigh/CD45RClow ratio, but not the percentage CD25+ cells, within the CD4 T cell compartment of naive rats is correlated with resistance to CsA-AI in these rat strains. We conclude that the relative presence of autoregulatory T cells with a CD45RClow T-helper cell phenotype may be a critical determinant in susceptibility to CsA-AI. PMID- 11869044 TI - Differences between two strains of myelin basic protein (MBP) TCR transgenic mice: implications for tolerance induction. AB - Experimental autoimmune encephalomyelitis (EAE) is mediated by CD4+ T cells which preferentially use the Vbeta8.2 TCR in response to myelin basic protein (MBP). Two strains of Tg mice (Valpha2.3/Vbeta8.2 and Valpha4/Vbeta8.2) have T cell receptors that recognize the NAc1-11 immunodominant epitope of MBP. We previously reported that oral administration of MBP protects both Valpha2.3/Vbeta8.2 and Valpha4/Vbeta8.2 mice from EAE; however, tolerance induction differs between strains and is dependent on the timing of oral antigen. Here we analyze the peripheral and gut-associated lymphoid tissue (GALT) environments of the two strains of Tg mice. Tg cells in the Peyer's patch (PP) but not the spleen of Valpha2.3/Vbeta8.2 mice demonstrate increased CD69 and decreased CD45RB relative to Valpha4/Vbeta8.2 mice. High levels of Th1 and Th2 cytokines, proliferative activity and CC chemokines (MCP-1) are observed in the periphery and GALT of Valpha2.3/Vbeta8.2 Tg mice. In contrast, more non-Tg CD4+ cells are seen in the PP of Valpha4/Vbeta8.2 mice. These studies suggest that activated Tg T cells and fewer potential regulatory cells in the PP of Valpha2.3/Vbeta8.2 Tg mice may influence oral tolerance. PMID- 11869046 TI - Stable GAD65 autoantibody epitope patterns in type 1 diabetes children five years after onset. AB - Autoantibodies to GAD65 (GAD65Ab) are prominent in type 1 diabetes. These autoantibodies may be present both years before and after the clinical diagnosis of type 1 diabetes and are widely used as a marker for the disease. Recently it has been demonstrated that progression to type 1 diabetes is accompanied by GAD65Ab epitope maturation. Here we examine whether autoantibody maturation processes also progress after the clinical diagnosis of type 1 diabetes. Antibody reactivity to GAD65, GAD67 and GAD65/67 fusion proteins was measured by radioimmunoassays in 62 children with type 1 diabetes. Samples were taken at diagnosis and five years later. While the overall GAD65Ab level declined over time, the epitope pattern was remarkably stable with no significant changes in binding pattern. Loss of GAD65Ab-positivity was associated with significantly lower GAD65Ab indices at diagnosis compared to patients' sera that remained GAD65Ab-positive. The decrease in GAD65Ab levels did not correlate to residual C peptide levels. Our data suggest that processes controlling GAD65Ab levels and epitope binding patterns remain stable during the first five years of type 1 diabetes. PMID- 11869047 TI - Predominantly recognized proinsulin T helper cell epitopes in individuals with and without islet cell autoimmunity. AB - Antibody response to insulin and to its precursor ProInsulin is associated with increased risk for type 1 diabetes (T1D), though little is known about T cell reactivity to this molecule. In the present study from peripheral blood mononuclear cells (PBMC), in vivo primed CD45RO+ memory T helper (Th) cells were enriched and their reactivity to eight overlapping ProInsulin peptides and to protein was analyzed. Individuals with high risk HLA-DRB1*04, DQB1*0302 alleles were investigated: relatives of patients with T1D having humoral markers of islet cell autoimmunity (autoantibody positive, Ab+; n=11), patients with T1D (n=8), and healthy control individuals (n=16). The ProInsulin epitope which was most frequently recognized in all the tested individuals was C-peptide (C) 18-A-chain (A)1. In Ab+ relatives the responses to this epitope and to two additional parts of the ProInsulin, B-chain (B) 11-C24 and C28-A21, was observed. In T1D patients who have already been treated with insulin, response to peptide B20-C4 and to the entire insulin molecule predominates. Our findings suggest that the spontaneous memory Th cell response to ProInsulin in individuals with high risk HLA alleles is predominantly directed to one epitope which maps to the central, C-peptide region. In individuals with humoral markers of islet cell autoimmunity and in patients with T1D, spread response to distinct ProInsulin regions was observed. PMID- 11869048 TI - ICA+ relatives with DQA1*0102/DQB1*0602 have expected 0602 sequence and DR types. AB - The HLA haplotype DQA1*0102/DQB1*0602 reportely confers protection from type 1 diabetes. DQA1*0102/DQB1*0602 is present in more than 7% of ICA positive relatives screened as part of the Diabetes Prevention Trial--type 1. The presence of autoantibodies in these subjects suggests that the mechanism that protects DQB1*0602 subjects from diabetes occurs after the disease process has been initiated. However, as previously suggested, the method used to type the DQB1*0602 alleles may have lacked the sensitivity to identify alleles similar, but not identical, to DQB1*0602. In addition unusual extended haplotypes may be presented that could help account for the presence of diabetes autoantibodies. We therefore sequenced and performed extended haplotyping on samples from ICA+ relatives with DQA1*0102/DQB1*0602. In this group, sequencing confirmed DQB1*0602 in 149/150, and 152/165 have the common DRB1*1501-DQB1*0602 haplotype. Thus, high resolution typing of class II alleles either by PCR-based oligotyping or nucleotide sequencing fail to indicate any unusual genetic characteristics about these antibody-positive relatives, of which few are expected to progress to clinical disease. PMID- 11869049 TI - Recombinant anti-idiotypic antibodies inhibit human natural anti-glycoprotein (GP)IIb/IIIa autoantibodies. AB - Anti-idiotypic antibodies (anti-Id) have been described against idiotypes expressed on various autoantibodies. Since an immunoregulatory effect has been postulated for anti-Id, modulation of the anti-Id response in autoimmune disease may be of interest. In chronic immune thrombocytopenic purpura (AITP), autoantibodies directed mainly against platelet membrane glycoprotein (GP) IIb/IIIa cause platelet destruction by Fc-mediated phagocytosis or by complement lysis. We have previously reported on the generation of two recombinant anti GPIIb/IIIa autoantibody fragments (PDG-X, PDG-B), that are specific for conformationally intact GPIIb/IIIa and inhibit binding of autoantibodies from patients with AITP. In the present study, we show that anti-GPIIb/IIIa specificities are not limited to a single individual by isolating five additional anti-GPIIb/IIIa autoantibody fragments from a second phagemid Fab library of an unrelated healthy donor. Using soluble Fab of PDG-X and PDG-B as antigens for panning Fab phagemid libraries from healthy human individuals, we isolated anti Id phage clones specific for PDG-X or PDG-B. In addition they inhibited the binding of PDG-X or PDG-B to GPIIb/IIIa. Amino acid sequence comparison between these specific antiId and GPIIb/IIIa was performed. Generation of these anti-Id directed against pathologically relevant anti-GPIIb/IIIa autoantibodies may represent a new suitable and specific therapeutic option for the treatment of antibody-mediated AITP. PMID- 11869051 TI - Haemobartonella felis: recent developments in diagnosis and treatment. AB - Haemobartonella felis is a pleomorphic uncultivated wall-less haemotrophic bacterial parasite. Phylogenetic analysis of 16S rRNA gene sequences from a number of isolates of H felis has demonstrated that these bacteria are most closely related to species in the genus Mycoplasma, and Haemobartonella and related organisms are currently being reclassified as Mollicutes. Diagnosis by cytological examination of blood smears has been problematic, but recent molecular studies have led to the development of sensitive polymerase chain reaction (PCR) assays for diagnosis. Such studies have also resulted in the recognition of two distinct strains of H felis, which are divided into different groups based on phylogenetic analysis. This evolutionary divergence between strains is accompanied by differences in pathogenecity. This review discusses new developments in the diagnosis and treatment of H felis, focusing on the use of, and interpretation of, PCR assays. PMID- 11869052 TI - Dietary chromium tripicolinate supplementation reduces glucose concentrations and improves glucose tolerance in normal-weight cats. AB - The effect of dietary chromium supplementation on glucose and insulin metabolism in healthy, non-obese cats was evaluated. Thirty-two cats were randomly divided into four groups and fed experimental diets consisting of a standard diet with 0 ppb (control), 150 ppb, 300 ppb, or 600 ppb added chromium as chromium tripicolinate. Intravenous glucose tolerance, insulin tolerance and insulin sensitivity tests with minimal model analysis were performed before and after 6 weeks of feeding the test diets. During the glucose tolerance test, glucose concentrations, area under the glucose concentration-time curve, and glucose half life (300 ppb only), were significantly lower after the trial in cats supplemented with 300 ppb and 600 ppb chromium, compared with values before the trial. Fasting glucose concentrations measured on a different day in the biochemistry profile were also significantly lower after supplementation with 600 ppb chromium. There were no significant differences in insulin concentrations or indices in either the glucose or insulin tolerance tests following chromium supplementation, nor were there any differences between groups before or after the dietary trial.Importantly, this study has shown a small but significant, dose dependent improvement in glucose tolerance in healthy, non-obese cats supplemented with dietary chromium. Further long-term studies are warranted to determine if the addition of chromium to feline diets is advantageous. Cats most likely to benefit are those with glucose intolerance and insulin resistance from lack of exercise, obesity and old age. Healthy cats at risk of glucose intolerance and diabetes from underlying low insulin sensitivity or genetic factors may also benefit from long-term chromium supplementation. PMID- 11869053 TI - Gastroduodenal ulceration in cats: eight cases and a review of the literature. AB - Gastroduodenal ulceration (GU) and blood loss was diagnosed in eight cats and compared with 25 previously reported cases of feline GU. Cats with GU presented in a critical condition. Clinical signs consistent with gastrointestinal bleeding were infrequently identified although anaemia was a common finding. Non neoplastic causes of feline GU tended to have a shorter clinical course with ulcers confined to the stomach. Conversely, cats with tumour-associated GU usually had a more protracted clinical course, weight loss, and ulcers located in the stomach for gastric tumours and the duodenum for extra-intestinal tumours. In this series, definitive diagnosis was possible for cats with neoplasia (gastric tumours and gastrinoma), however, it was difficult to precisely identify the underlying aetiology in cats with non-neoplastic GU. Prompt stabilisation with a compatible blood transfusion, surgical debridement or resection, antibiotic and antiulcer therapy, and treatment of the underlying disease, if identified, was successful in the majority of cases. The prognosis for cats with appropriately managed GU depended on the underlying aetiology, but even cats with neoplasia could be successfully palliated for prolonged periods. PMID- 11869055 TI - Primary right atrium haemangiosarcoma in a cat. AB - Primary cardiac neoplasms are rare in animals. Hemangiosarcoma is the most common primary cardiac tumour in the dog, but has not been reported in the cat. We describe the clinical signs, the gross examination, and the morphological and immunohistochemical features of a haemangiosarcoma in the right atrium of a 13 year-old domestic shorthair cat. In addition, a distant metastasis was found in the liver. PMID- 11869054 TI - Feline leprosy: two different clinical syndromes. AB - Feline leprosy refers to a condition in which cats develop granulomas of the subcutis and skin in association with intracellular acid-fast bacilli that do not grow on routine laboratory media. In this study, the definition was extended to include cases not cultured, but in which the polymerase chain reaction (PCR) identified amplicons characteristic of mycobacteria. Tissue specimens from 13 such cases from eastern Australia were obtained between 1988 and 2000. This cohort of cats could be divided into two groups on the basis of the patients' age, histology of lesions, clinical course and the sequence of 16S rRNA PCR amplicons. One group consisted of four young cats (less than 4 years) which initially developed localised nodular disease affecting the limbs. Lesions progressed rapidly and sometimes ulcerated. Sparse to moderate numbers of acid fast bacilli were identified using cytology and/or histology, typically in areas of caseous necrosis and surrounded by pyogranulomatous inflammation. Organisms did not stain with haematoxylin and ranged from 2 to 6 microm (usually 2 to 4 microm). Mycobacterium lepraemurium was diagnosed in two cases based on the sequence of a 446 bp fragment encompassing the V2 and V3 hypervariable regions of the 16S rRNA gene a different sequence was obtained from one additional case, while no PCR product could be obtained from the remaining case. The clinical course was considered aggressive, with a tendency towards local spread, recurrence following surgery and development of widespread lesions over several weeks. The cats resided in suburban or rural environments. A second group consisted of nine old cats (greater than 9 years) with generalised skin involvement, multibacillary histology and a slowly progressive clinical course. Seven cats initially had localised disease which subsequently became widespread, while two cats allegedly had generalised disease from the outset. Disease progression was protracted (compared to the first group of cats), typically taking months to years, and skin nodules did not ulcerate. Microscopically, lesions consisted of sheets of epithelioid cells containing large to enormous numbers of acid-fast bacilli 2 to 8 microm (mostly 4 to 6 microm) which stained also with haematoxylin. A single unique sequence spanning a 557 bp fragment of the 16S rRNA gene was identified in six of seven cases in which it was attempted. Formalin-fixed paraffin-embedded material was utilised by one laboratory, while fresh tissue was used in another. The same unique sequence was identified despite the use of different primers and PCR methodologies in the two laboratories. A very slow, pure growth of a mycobacteria species was observed on Lowenstein Jensen medium (supplemented with iron) and semi-solid agar in one of three cases in which culture was attempted at a reference laboratory. Affected cats were domicile in rural or semi-rural environments. These infections could generally be cured using two or three of rifampicin (10-15 mg/kg once a day), clofazimine (25 to 50 mg once a day or 50 mg every other day) and clarithromycin (62.5 mg per cat every 12 h). These findings suggest that feline leprosy comprises two different clinical syndromes, one tending to occur in young cats and caused typically by M lepraemurium and another in old cats caused by a single novel mycobacterial species. PMID- 11869056 TI - Acute intervertebral disc extrusion in a cat: clinical and MRI findings. AB - A 5 year old, neutered male, domestic shorthaired cat had acute left hemiparesis and Horner's syndrome. Magnetic resonance imaging (MRI) revealed a loss of the normal signal from the nucleus pulposus of the intervertebral disc at C3/4, narrowing of the ventral subarachnoid space and slight dorsal displacement of the spinal cord and a focal hyperintense lesion affecting the left side of the spinal cord at the same level. The presumptive diagnosis was focal spinal cord oedema associated with intervertebral disc extrusion. A traumatic aetiology was suspected. The cat was treated conservatively and improved gradually over a period of 6 months. PMID- 11869058 TI - The causes of fever in children attending hospital in the north of England. AB - OBJECTIVES: Fever is a common symptom in children presenting to casualty. Identifying the seriously ill is difficult. Previous studies, mainly from North America, suggest that symptoms, signs and simple investigations may help to do this. The aim of the present study was to assess the causes of fever and identify clinical and laboratory features suggesting serious disease in U.K. children presenting to hospital with temperatures >or=38 degrees C. METHODS: All children with a temperature of >or=38 degrees C seen in two hospitals between August and October 1999. RESULTS: One hundred and forty one children between 8 days and 16 years of age were studied, 64% male, 55% aged under 2 years. Eighty three percent had temperatures between 38 and 39 degrees C. Ninety six percent were casualty or GP referrals and 4% were tertiary referrals. Twenty nine percent (41/141) had serious disease but microbiologically or radiologically proven in only 22% (31/141); pneumonia (nine), meningitis (seven), sepsis (five), urinary tract infection (five), brain abscess (two), toxic shock syndrome (one), appendicitis (one), ischiorectal abscess (one). Forty two percent (5/12) of microbiologically proven meningitis and sepsis and 36% (8/22) of all meningitis and sepsis were meningococcal. Seventy one percent had non-serious diseases. In cases of serious disease the temperature was >39 degrees C in 15% (sensitivity: 14%, specificity: 82%, PPV: 25%). Poor feeding and restlessness predicted serious disease with a sensitivity of 78% and 76%, respectively. Full blood count (FBC) was taken in 50% of patients on admission; in 44% of serious and 24% of non-serious diseases WBC was between 5000 and 15,000/mm(3) and WBC >or=15,000/mm(3) was seen in 39% of serious diseases (sensitivity:10%, specificity: 95%, PPV: 44%). CONCLUSIONS: One out of three of children referred with fever had a serious disease. Degree of temperature and WBC count were poor predictors of serious disease. Interestingly, poor feeding and restlessness were more sensitive predictors, suggesting high fever and WBC count can not replace clinical assessment of the child with a temperature. PMID- 11869060 TI - Treatment of resistant mycoplasma infection in immunocompromised patients with a new pleuromutilin antibiotic. AB - Patients with primary antibody deficiency (PAD) are prone to mycoplasma infection with unusual strains which may be resistant to conventional antibiotics. Mycoplasmas were isolated from the joint fluid (Ureaplasma urealyticum) of two PAD patients with arthritis and from the cerebral spinal fluid (Mycoplasma maculosum) in one with meningitis, the latter probably originating from the patient's dog. Combinations of doxycycline and quinolones or macrolides failed to clear the infections, but after demonstrating in-vitro sensitivity to the pleuromutilin, Econor, for two of the isolates, all three patients responded to oral treatment with Econor. The infection was completely eradicated in two patients, with the emergence of a resistant strain in the third. Mycoplasma infection should be considered in PAD patients with unexplained sepsis. Pleuromutilins such as Econor are powerful new anti-mycoplasmal agents which provide an additional therapeutic option when patients fail to respond to conventional antibiotics. PMID- 11869059 TI - A prospective study of AIDS-associated cryptococcal meningitis in Thailand treated with high-dose amphotericin B. AB - OBJECTIVE: To assess kinetic of cryptococci in the cerebrospinal fluid (CSF) and outcome of AIDS-associated cyptococcal meningitis after high-dose amphotericin B. PATIENTS AND METHODS: A prospective study involving Thai adults (n=106) with cryptococcal meningitis associated with AIDS was conducted to determine the kinetic of cryptococci in CSF and prognostic factors affecting survival after high-dose amphotericin B (0.7 mg/kg/day) followed by oral azole treatment. Cerebrospinal fluids were collected for cryptococcal count and culture at weekly intervals for at least 2 weeks or until CSF cultures were negative for cryptococci. All patients were followed monthly for 1 year or until death in order to detect relapse or occurrence of any other opportunistic infection. RESULTS: A total of 106 AIDS patients with cryptococcal meningitis were enrolled. The geometric mean (range) total and viable cryptococcal counts in CSF on admission were 430,000 (1000 to 3.4 x 10(7)) and 31,000 (10 to 1.4 x 10(7)) per ml, respectively. Both total and viable cryptococcal counts declined monoexponentially with an elimination half life of 4 days. The cumulative CSF yeast clearance rates were 38% and 56% at 2 and 4 weeks, respectively. Early death was associated significantly with previous history of weight loss [relative risk (RR)=2.2; 95% CI, 1.2-3.9], Glasgow Coma Score <13 (RR=2.33; 95% CI, 1.55 3.50), and hypoalbuminaemia (P<0.001). Later mortality was associated delayed CSF yeast clearance (RR=3.6; 95% CI, 1.9--6.4) and relapse (RR=3.9; 95% CI, 1.4 10.8). CONCLUSION: High-dose amphotericin B was not as effective as previously thought. Cumulative mortality at 2 weeks, 4 weeks and 1 year were 16%, 24% and 76%, respectively. PMID- 11869061 TI - Comparative analysis of total and integrated HIV-1 DNA in peripheral CD4 lymphocytes and monocytes after long treatment with HAART. AB - OBJECTIVE: To determine the level of total and integrated HIV-1 DNA load in CD4 lymphocytes and monocytes of patients undergoing HAART treatment for at least 2 years. METHODS: CD4 lymphocytes were isolated by subjecting monocyte-depleted blood samples to immune-purging carried out with M-450 Dynabeads. Monocytes were separated by blood through a combined procedure of cell adherence to dishes and complement induced immune lysis with anti-CD3 Mab. HIV DNA in CD4 lymphocytes and monocytes was quantified by polymerase chain reaction (PCR) based limit dilution assay with two PCR protocols, specific for total (LTR PCR) and integrated (Alu PCR) forms of HIV DNA. The replication competence of the provirus harboured in monocyte-depleted peripheral blood mononuclear cells (PBMC) and adherent monocytes was assayed by measuring HIV-1 p24 antigen produced by in-vitro cultures established with these cells. RESULTS: The CD4 lymphocytes of all patients contained a consistent number of HIV DNA copies. Most patients were also positive for HIV DNA in monocytes. The Alu PCR analysis detected, integrated provirus in CD4 lymphocytes of 9 patients and in the monocytes of only three. Four patients had replication-competence virus in their PBL. The monocytes of all patients did not produce virus in vitro. CONCLUSION: The HIV infection of CD4 lymphocytes and monocytes is maintained even after HAART related, apparent, and durable suppression of viral replication. We suggest that the viral persistent infection of monocytes may play a role in maintaining the residual HIV activity found in patients undergoing HAART. PMID- 11869062 TI - Toxic epidermal necrolysis induced by nevirapine therapy: description of two cases and review of the literature. AB - We describe two cases of toxic epidermal necrolysis developed during an antiretroviral therapy regimen containing nevirapine. It seems likely that the poor adherence to the dose escalation regimen of nevirapine has caused this life threatening disease. A complete and written information on the scheduled antiretroviral therapy is mandatory, above all for individuals coming from developing countries where language barriers have not yet been successfully overcome. PMID- 11869063 TI - Ventricular patch endocarditis caused by Propionibacterium acnes: advantages of gallium scanning. AB - Propionibacterium acnes is a weakly pathogenic commensal of the skin. When isolated from blood cultures it is often considered a contaminant. However, P. acnes may be responsible for severe infections and its role in certain cases of infectious endocarditis has now been definitely established.(1) We report a case of endocarditis due to P. acnes stemming from a ventricular patch and revealed by a gallium 67 scan. PMID- 11869064 TI - Tuberculosis transmitted through transplantation. AB - Tuberculosis in solid organ transplant recipients is associated with relatively high morbidity and mortality and is often extra-pulmonary. Reactivation of dormant infection is the usual mode of acquisition with donor and nosocomial transmission occurring infrequently. We report two cases of probable donor transmitted extra-pulmonary infection where both isolates of Mycobacterium tuberculosis proved to be indistinguishable using hemi-nested inverse PCR of the IS 6110 region. PMID- 11869065 TI - Pyoderma gangrenosum complicating a spider bite in the UK. PMID- 11869066 TI - Peritoneal tuberculosis and elevated serum CA 125 levels: recognizing the association is important. PMID- 11869068 TI - Functional anatomy of articular cartilage under compressive loading Quantitative aspects of global, local and zonal reactions of the collagenous network with respect to the surface integrity. AB - OBJECTIVE: To assess the influence of local compressive loading on the arrangement of the collagenous fibers in intact articular cartilage. To quantitate the zonal deformation of intact cartilage under load. To analyse the influence of removal of the tangential zone on the load-induced changes. MATERIALS AND METHODS: 380 cylinder shaped cartilage-on-bone samples (d=7 mm) were harvested from 20 bovine femoral heads. In 120 of them the tangential zone was removed. All samples were loaded for 20 min by 0.42 MPa or 0.98 MPa. After proteoglycan extraction, fixation in 4% formalin, dehydration by increasing concentrations of acetone, critical point drying, freeze-fracturing and gold coating the samples were analysed by scanning-electron-microscopy. RESULTS: Fiber bulging away from the center of load occurred in an area larger than the directly loaded one and its extent increased parallel to loading (P< 0.01). Crimp was seen only under the indenter and spread with increasing load from the intermediate zone into the tangential zone and radial zone. The absolute height of tangential zone and intermediate zone together remained constant under all loading situations at the costs of the radial zone. All changes due to loading were fully reversible. Removal of the tangential zone reduced the area of bulging (P< 0.01) but markedly increased the amount of crimp. Overall radial strain was not altered, but overall superficial tangential strain was increased by up to 20% (P< 0.01) and high peaks in the local distribution of superficial tensile strain developed. CONCLUSIONS: The collagenous architecture is a dynamic property of the articular cartilage adapting to its respective loading situation. Crimp reflects local compressive strain. Under compressive loading larger portions of cartilage than the directly loaded areas are functionally included in the process of load transmission. During this process the tangential zone and the intermediate zone form a common functional unit providing a high degree of fiber cross-linkage as a possible mechanism to increase zonal compressive stiffness. Removal of the tangential zone seems to impair distribution of a locally applied compressive load sideways and leads to a reduced cartilage volume included in the process of load transmission. An intact tangential zone contributes to prevent peaks of surface tensile strain. PMID- 11869069 TI - Peroxisome proliferator-activated receptor gamma activators inhibit MMP-1 production in human synovial fibroblasts likely by reducing the binding of the activator protein 1. AB - OBJECTIVE: To investigate the expression and activity of PPARgamma in human synovial fibroblasts and the effects of PPARgamma agonists on the expression of MMP-1. The molecular mechanisms by which PPARgamma agonists modulate MMP-1 expression were also examined. METHODS: PPARgamma expression and activity were measured using reverse-transcription polymerase chain reaction (RT-PCR) and transient transfection assays. Human synovial fibroblasts were cultured with IL 1beta in the absence or presence of PPARgamma activators, and the expression and production of MMP-1 were evaluated by Northern blot and ELISA, respectively. The effect of 15d-PGJ(2) on MMP-1 promoter activation was analysed in transient transfection experiments, while electrophoretic mobility shift assays were performed to study the binding activity of the transcription factor AP-1. RESULTS: PPARgamma was expressed and transcriptionally functional in human synovial fibroblasts. PPARgamma activators (15d-PGJ(2) and BRL 49653) inhibited IL-1beta-induced MMP-1 synthesis in a dose-dependent manner. Similarly, both activators inhibited IL-1-induced MMP-1 mRNA expression. Activation of the human MMP-1 promoter was also attenuated by 15d-PGJ(2), indicating that the inhibitory effect of 15d-PGJ(2) occurs at the transcriptional level. Interestingly, 15d PGJ(2) reduced both basal and IL-1beta-induced AP-1 binding activity. CONCLUSIONS: These data indicate that PPARgamma agonists inhibit MMP-1 gene expression by transcriptional mechanisms, and suggest that they may be useful in reducing joint tissue destruction. PMID- 11869070 TI - Vital marking of articular chondrocytes by retroviral infection using green fluorescence protein. AB - OBJECTIVE: One of the main open questions in chondrocyte transplantation is the fate of the implanted cells in vivo. We intended to establish prerequisites for such studies in animal models and to show the feasibility of this approach in rabbits. Isolated articular chondrocytes were retrovirally marked using green fluorescence protein (GFP) as a cell-specific marker in order to allow an in vivo follow-up of these cells. METHODS: Chondrocytes from rabbits, sheep, cattle and humans were isolated and infected with murine leukemia virus-derived retroviruses carrying the GFP gene. The influence of the host range of three packaging cell lines (PA317, PT67, PG13), start cell concentrations, number of cell passages and number of infection cycles on the efficiency of infection was investigated. Stability of GFP expression was followed by FACS analysis, confocal imaging and fluorescence microscopy. For in vivo follow-up of GFP expression we used marked allogeneic chondrocyte populations grown on scaffold material and implanted them into full-thickness defects in knee joints of rabbits. RESULTS: Retroviruses from all three packaging cell lines were able to infect rabbit and human chondrocytes, whereas only retroviruses released from PG13 cells were able to infect sheep and bovine chondrocytes efficiently. Optimization of the infection with these viruses resulted in efficiencies of 60-90% GFP-expressing chondrocytes. Populations of 100% marked chondrocytes were obtained by cell sorting. GFP expression stability of such marked chondrocyte populations was followed in monolayer culture and in 3 D culture on different scaffold materials. The expression of GFP was stable on all tested materials for at least 4 weeks. In monolayer culture GFP expression was stable for more than 8 months. In vivo, we observed stable GFP expression in the transplants during a four-week time course. CONCLUSION: Retroviral GFP gene transfer led to long-term expression in chondrocytes from rabbits, sheep, cattle and humans. Transgene expression and the number of implanted chondrocytes remain stable for at least 4 weeks in vivo. This method permits a rapid monitoring of chondrocytes and provides a basis for following the fate of these cells in vivo. PMID- 11869071 TI - Dietary vitamins and selenium diminish the development of mechanically induced osteoarthritis and increase the expression of antioxidative enzymes in the knee joint of STR/1N mice. AB - OBJECTIVE: To study the influence of dietary vitamins and selenium on mechanically-induced osteoarthritis (OA) and the expression of antioxidative enzymes in male STR/1N and Balb/c mice. Male STR/1N mice are prone to develop OA caused by a varus deformity-induced mechanical overload of the medial tibial plateau. METHODS: After 12 months of feeding (special diet supplemented with the vitamins E, C, A, B6, B2, and selenium) serial histological sections of the knee joints were evaluated for development of osteoarthritic changes (grade 0-4). Serum glutathione peroxidase activity (GSH-px) was measured photometrically. Expression of antioxidative enzymes was demonstrated by immunohistochemistry. RESULTS: All control STR/1N mice showed OA lesions (grade 3-4) while the special diet decreased OA incidence significantly down to approximately 65% (mostly grade 2). Even in Balb/c mice the incidence was decreased by the special diet from approximately 21% (control animals; grade 1) to approximately 14%. Serum GSH-px activity increased diet-dependently in both mouse strains but was generally higher in Balb/c mice. In both mouse strains the special diet increased the expression of GSH-px and Cu/Zn-SOD in articular cartilage while there was no expression of Mn-SOD. There was also a special diet-dependent increase in expression of GSH-px in the synovium of both mouse strains while an increase in expression of Mn-SOD and Cu/Zn-SOD could only be seen in the synovium of STR/1N mice. CONCLUSIONS: A diet supplemented with vitamins/selenium might be important in prevention or therapy of mechanically induced OA. We hypothesize that free oxygen radical species might be involved in the mechanical induction of OA. PMID- 11869072 TI - Putative role of lysyl hydroxylation and pyridinoline cross-linking during adolescence in the occurrence of osteoarthritis at old age. AB - OBJECTIVE: The collagen network in human articular cartilage experiences a large number of stress cycles during life as it shows hardly any turnover after adolescence. We hypothesized that, to withstand fatigue failure, the physical condition of the collagen network laid down at adolescence is of crucial importance for the age of onset of osteoarthritis (OA). METHODS: We have compared the lysyl hydroxylation level and pyridinoline cross-link level of the collagen network of degenerated (DG) cartilage of the femoral knee condyle (representing a preclinical early stage of OA) with that of normal cartilage from the contralateral knee. The biological age of the collagen network was determined by means of pentosidine levels. For each donor, collagen modifications of normal cartilage were compared with DG cartilage that showed no significant remodeling of the collagen network (as evidenced by identical pentosidine levels). RESULTS: DG cartilage contained significantly more hydroxylysine residues per collagen molecule in comparison with healthy cartilage from the same donor, both in the upper and lower half (the region near the articular surface and adjacent to bone, respectively). In addition, a significantly higher level of pyridinoline cross linking was observed in the upper half of DG cartilage. Considering the biological age of the collagen network, the changes observed in DG cartilage must have been present several decades before cartilage became degenerated. CONCLUSIONS: The data suggest that high levels of lysyl hydroxylation and pyridinoline cross-linking result in a collagen network that fails mechanically in long term loading. Areas containing collagen with low hydroxylysine and pyridinoline levels are less prone to degeneration. As such, this study indicates that post-translational modifications of collagen molecules synthesized during adolescence are causally involved in the pathogenesis of OA. PMID- 11869073 TI - Expression of beta1 integrins during periosteal chondrogenesis. AB - OBJECTIVE: The interactions between integrins and extracellular matrix proteins are known to modulate cell behavior, and may be involved in regulating cartilage formation and repair. The purpose of this study was to determine the patterns and localization of expression of the beta1 integrins during cartilage formation by periosteum, which is used to repair articular cartilage. DESIGN: Periosteal explants from 2-month-old rabbit medial proximal tibiae were cultured in agarose suspension for 0 to 6 weeks, with 10 ng/ml transforming growth factor-beta1 added for the first 2 days of culture. Integrin expressions were measured by reverse transcriptase-polymerase chain reaction (RT-PCR) and localized by immunohistochemistry. RESULTS: Normal periosteum expressed the alpha1, alpha3, alpha5, beta1 subunits at low levels, and the proteins for all but the alpha3 subunits were identified by immunohistochemistry in the periosteum. Significant two- to five-fold up-regulation of the mRNA expression of the alpha1, alpha3, alpha5 and beta1 integrin subunits during the early proliferative stage of chondrogenesis was observed. The initial change was a five-fold increase in alpha5 expression on day 2 and a two-fold increase in alpha3 expression. On day 5, alpha1 expression was up-regulated (four-fold). beta1 expression was broadly up-regulated (three to four-fold) from day 5 to 14. In the early stage of chondrocyte differentiation, after day 14, alpha1 expression was down-regulated, while there was upregulation of alpha3 (three-fold), alpha5 (three-fold) and beta1 (four-fold) expressions. Thereafter, alpha1 expression was down-regulated, while alpha3, alpha5 and beta1 expressions were up-regulated again during matrix synthesis. Immunohistochemistry confirmed this late decrease in alpha1 levels and increase in alpha3, alpha5 and beta1 levels in chondrocytes. CONCLUSIONS: These observations indicate that the beta1 integrins play an important role in the process of chondrogenesis in periosteum. PMID- 11869074 TI - The canine 'groove' model, compared with the ACLT model of osteoarthritis. AB - OBJECTIVE: The frequently used anterior cruciate ligament transection (ACLT) model of osteoarthritis (OA) in the dog, makes use of a permanent trigger (joint instability) for inducing degenerative changes. The present study evaluates a canine model of degenerative cartilage damage, mimicking OA, which is induced without making use of permanent joint instability. METHODS: The articular cartilage of the weight-bearing areas of the femoral condyles in one knee of ten beagle dogs was damaged by making grooves, without damaging the subchondral bone. Surgery was followed by 10 weeks intensified loading of the affected joint. Subsequently, joint damage and inflammation were evaluated. The effects were compared with those of the ACLT model. RESULTS: Histological analysis showed chondrocyte clusters around cartilage lesions and moderate loss of proteoglycans in the 'groove' model. Synovial inflammation was mild. Biochemical analysis of cartilage showed changes in matrix proteoglycan turnover, proteoglycan content, and collagen damage, all characteristics of OA. Synovial fluid MMP-1, -3 and -13 activity was enhanced. Changes were found in condyles and plateau, were similar for all animals tested, and were similar to the changes observed in the ACLT model. CONCLUSION: The presently described canine 'groove' model shows characteristics identical to those seen in the ACLT model but differs in a way that the changes are induced without joint instability. The latter is expected to make the 'groove' model more sensitive to treatment. PMID- 11869075 TI - Increased cellular infiltrate in inflammatory synovia of osteoarthritic knees. AB - OBJECTIVE: To determine the nature and origin of synovial inflammation in knees with osteoarthritis (OA). METHOD: Synovial samples were obtained from 21 medial compartmental knee OAs from 19 patients. First, using 11 medial knee synovial samples from 9 patients, a quantitative estimation of synovitis was made with both ordinary and immunohistochemical staining. Second, from the other 10 knees, tissue samples were taken from both the medial and the lateral compartments to quantify cells that infiltrated into the synovium. Medial synovium was immunostained using antibodies to anti-type II collagen, CD68, CD2, CD4, CD8, CD15, CD19, CD25, HLA-DR, CD1a and LN5. The lateral synovium was immunostained with anti-type II collagen, CD68, HLA-DR and CD4 antibody as a control. RESULT: Denatured cartilaginous detritus was found captured by synovial lining cells with a strong immunoreactivity to CD68 antibody, and whose phagocytic potential was activated. The number of anti-type II collagen-positive fragments in the medial compartment of the knee was larger than that found in the lateral compartment. Moreover, the population of CD68-positive cells in synovial tissue and HLA-DR positive cells in the lining layer was larger in the medial compartment than in the lateral compartment. The number of CD4-positive cells (defined as helper/inducer T lymphocytes) was greater in medial synovium than in lateral synovium. CONCLUSION: Overall, this study strongly supports the concept that the synovitis observed in patients with knee OA might be induced by an immunological mechanism involving, to some extent, a macrophage/helper T cell interaction. PMID- 11869076 TI - Diffraction-enhanced X-ray imaging of articular cartilage. AB - OBJECTIVE: To introduce a novel X-ray technology, diffraction-enhanced X-ray imaging (DEI), in its early stages of development, for the imaging of articular cartilage. DESIGN: Disarticulated and/or intact human knee and talocrural joints displaying both undegenerated and degenerated articular cartilage were imaged with DEI. A series of three silicon crystals were used to produce a highly collimated monochromatic X-ray beam to achieve scatter-rejection at the microradian level. The third crystal (analyser) was set at different angles resulting in images displaying different characteristics. Once the diffraction enhanced (DE) images were obtained, they were compared to gross and histological examination. RESULTS: Articular cartilage in both disarticulated and intact joints could be visualized through DEI. For each specimen, DE images were reflective of their gross and histological appearance. For each different angle of the analyser crystal, there was a slight difference in appearance in the specimen image, with certain characteristics changing in their contrast intensity as the analyser angle changed. CONCLUSIONS: DEI is capable of imaging articular cartilage in disarticulated, as well as in intact joints. Gross cartilage defects, even at early stages of development, can be visualized due to a combination of high spatial resolution and detection of X-ray refraction, extinction and absorption patterns. Furthermore, DE images displaying contrast heterogeneities indicative of cartilage degeneration correspond to the degeneration detected by gross and histological examination. PMID- 11869077 TI - Comparison of collagenase-cleaved articular cartilage collagen in mice in the naturally occurring STR/ort model of osteoarthritis and in collagen-induced arthritis. AB - OBJECTIVE: The STR/ort mouse develops a naturally occurring osteoarthritis of the femorotibial joint that provides a model with which to establish the time course of biochemical changes taking place in articular cartilage in the disease. Our objective was to define the onset, location and progression of type II collagen cleavage by collagenase in the tibial cartilage of the STR/ort mouse. For comparison, cartilage collagen cleavage was also studied in collagen-induced arthritis in DBA mice. DESIGN: STR and control CBA mice aged 6-45 weeks were examined. DBA/1 mice were studied 2 and 3 weeks after initiating collagen-induced arthritis. Collagen cleavage was detected by immunolocalization using the antibody COL2-3/4Cshort which recognizes a carboxy terminal neoepitope created by collagenase cleavage of type I and II collagens. RESULTS: No COL 2-3/4Cshort immunostaining was observed in the intact cartilage of healthy young or old mice. The earliest detectable collagen degradation occurred at the cartilage surface coincident with the appearance of surface roughening. As fibrillations developed, further collagen degradation was evident around the edge of the lesion and in adjacent extracellular matrix. In contrast, staining was observed throughout the cartilage matrix in type II collagen-induced arthritis prior to the development of histopathological lesions. CONCLUSION: No evidence was found for collagen cleavage in intact/pre-lesional cartilage from STR/ort mice. Local collagen cleavage was, however, clearly associated with very early histopathological lesions and immunostaining with COL 2-3/4Cshort increased with progression of the latter. In contrast, type II collagen cleavage occurs throughout the articular cartilage at an early stage in collagen-induced arthritis. PMID- 11869078 TI - Development and regulation of osteophyte formation during experimental osteoarthritis. AB - OBJECTIVES: Osteophytes represent areas of new cartilage and bone formation in human and experimentally induced osteoarthritis (OA). The present study addressed the production of nitric oxide (NO), vascular endothelial growth factor (VEGF) and the occurrence of apoptosis during osteophyte formation. DESIGN: Osteophytes in the knee joint of rabbits that developed OA-like lesions following anterior cruciate ligament transection (ACLT) were analysed by histology and immunohistochemistry for NO production, and the presence of VEGF. TUNEL was used to detect DNA fragmentation. RESULTS: At the joint margins in the interface between cortical bone marrow and periosteal lining growth plate-like formations were detectable as early as 4 weeks after ACLT. By 12 weeks after ACLT osteophytes were visible in 100% of femoral condyles and tibial plateaus. Discrete areas with proliferating chondrocytes, hypertrophic chondrocytes, calcified matrix and vascular invasion were observed. VEGF immunoreactivity was most prominent in hypertrophic chondrocytes 9 weeks after ACLT. Nitrotyrosine immunoreactivity was detected in endothelial cells and in some hypertrophic chondrocytes in the calcified zone 4 weeks after ACLT. After 8 and 12 weeks, positive cells were detected in the hypertrophic and calcified zone. TUNEL positive cells were seen in blood vessels, and among hypertrophic chondrocytes adjacent to the blood vessels 4 weeks after ACLT. The proliferative zone, pre hypertrophic zone and hypertrophic zone showed only a few TUNEL positive cells. In contrast, 8 weeks and 12 weeks after ACLT, most hypertrophic chondrocytes, but few proliferative chondrocytes showed DNA fragmentation. CONCLUSIONS: Hypertrophic chondrocytes in osteophytes express VEGF and this can promote vascular invasion of cartilage. The presence of TUNEL-positive cells shows a similar distribution as nitrotyrosine immunoreactivity during all phases of osteophyte development, suggesting that NO production and chondrocyte death are related events in osteophyte formation. PMID- 11869079 TI - Control of extracellular matrix homeostasis of normal cartilage by a TGFbeta autocrine pathway. Validation of flow cytometry as a tool to study chondrocyte metabolism in vitro. AB - OBJECTIVE: To validate flow cytometry as an experimental technique for the study of the homeostasis of the extracellular matrix (ECM) of human articular cartilage. METHODS: Given the established insights in the relation between the transforming growth factor (TGF)-beta type II Receptor (TGF-betaRII)/TGF-beta auto/paracrine pathway, the intracellular levels of matrix metalloproteinases (MMPs) and their natural inhibitors (TIMPs), and the accumulation of ECM molecules in the ECM of articular cartilage, this metabolic pathway was used as a reference model to fulfill the objective. Chondrocytes were liberated from visually intact femoral condyle cartilage and cultured in gelled agarose to maintain their differentiated phenotype. After 2 weeks of culture, the chondrocytes were isolated from the agarose and flow cytometry was used to analyse the expression of TGF-betaRII on the plasmamembrane, the expression of TGFbeta1, MMP-1, MMP-3, TIMP-1 and TIMP-3 inside the cells, as well as the amounts of aggrecan, type II collagen and hyaluronan in the cell-associated matrix (CAM). The expression of the different substances was analysed with flow cytometry and reported as mean fluorescence intensity (MFI), which is due to the binding of FITC-labeled antibodies to their specific antigens. In addition, the effects of exogenous TGFbeta1 on the expression of these proteins was investigated on chondrocytes cultured in serum-free media. Enzyme Linked Immunosorbent Assay (ELISA) was performed to evaluate the MMP-1, MMP-3, TIMP-1 and MMP-1/TIMP-1 complex in the culture medium collected after the last 3 days of the culture period. The correlations between the data were analysed with the Spearman's test. RESULTS: Exogenous TGF-beta1 increased the accumulation of aggrecan and hyaluronan in the CAM of chondrocytes and down-regulated the intracellular levels of MMP-1 and -3. TIMP-1 and -3 were increased after exposure to TGF-beta1. Baseline expression of TGF-betaRII on the plasmamembrane of normal human articular chondrocytes significantly correlated with the intracellular levels of TGFbeta1, TIMP-1 and TIMP-3. TGFbeta1 was correlated with TIMP-1, TIMP 3 and MMP-1. Aggrecan in the CAM was inversely correlated with the ratio of MMP-1 to TIMPs. In addition, there were correlations between TIMP-1 and TIMP-3, aggrecan and hyaluronan. ELISA also revealed the correlation between MMP-1 and TIMP-1 secreted by the chondrocytes into the nutrient medium. MMP-1/TIMP-1 complex was hardly found in the medium. CONCLUSIONS: Some aspects of ECM metabolism of normal cartilage were evaluated by flow cytometry. Chondrocytes from normal human cartilage, when cultured in gelled agarose, showed correlations between the expression of TGF-betaRII/TGF-beta1 and the intracellular levels of TIMPs, indicating that TGF-beta autocrine pathway may contribute to homeostasis of the ECM in the normal cartilage. The relations between MMPs, TIMPs and the ECM molecules support that a physiological balance between MMPs and TIMPs results in a well-controlled matrix turnover in normal cartilage. PMID- 11869080 TI - Human autologous culture expanded bone marrow mesenchymal cell transplantation for repair of cartilage defects in osteoarthritic knees. AB - OBJECTIVE: There is no widely accepted method to repair articular cartilage defects. Bone marrow mesenchymal cells have the potential to differentiate into bone, cartilage, fat and muscle. Bone marrow mesenchymal cell transplantation is easy to use clinically because cells can be easily obtained and can be multiplied without losing their capacity of differentiation. The objective of this study was to apply these cell transplantations to repair human articular cartilage defects in osteoarthritic knee joints. DESIGN: Twenty-four knees of 24 patients with knee osteoarthritis (OA) who underwent a high tibial osteotomy comprised the study group. Adherent cells in bone marrow aspirates were culture expanded, embedded in collagen gel, transplanted into the articular cartilage defect in the medial femoral condyle and covered with autologous periosteum at the time of 12 high tibial osteotomies. The other 12 subjects served as cell-free controls. RESULTS: In the cell-transplanted group, as early as 6.3 weeks after transplantation the defects were covered with white to pink soft tissue, in which metachromasia was partially observed. Forty-two weeks after transplantation, the defects were covered with white soft tissue, in which metachromasia was observed in almost all areas of the sampled tissue and hyaline cartilage-like tissue was partially observed. Although the clinical improvement was not significantly different, the arthroscopic and histological grading score was better in the cell-transplanted group than in the cell-free control group. CONCLUSIONS: This procedure highlights the availability of autologous culture expanded bone marrow mesenchymal cell transplantation for the repair of articular cartilage defects in humans. PMID- 11869081 TI - The prevalence of nitric oxide in apoptotic chondrocytes of osteoarthritis. AB - OBJECTIVE: Apoptosis appears to be a significant mechanism of chondrocyte death in osteoarthritis (OA). There is increasing evidence that nitric oxide (NO) may be the inducing signal for apoptosis, but no study has definitively shown an association between the two in vivo. In this study, sections of osteoarthritic cartilage were double stained for the presence of apoptosis and NO to test the hypothesis that NO is the inducer of apoptosis in arthritis. DESIGN: Sections of osteoarthritic cartilage obtained during total knee arthroplasty were stained for apoptosis with terminal transferase-mediated dUTP nick end labeling (TUNEL). The sections were then stained for nitrotyrosine (a marker of NO production) by immunohistochemistry. The prevalence of NO in cells positive for apoptosis and in cells negative for apoptosis was determined by fluorescent microscopy. RESULTS: The prevalence of NO in apoptotic cells was no different than in non-apoptotic cells, suggesting NO is not the initiating signal for apoptosis in vivo. CONCLUSIONS: The precipitating cause for apoptosis in arthritic chondrocytes has not yet been determined. The data from this study fail to support NO as the direct initiating signal. NO synthase inhibitors may still be useful in the treatment of OA by blocking the catabolic activities of NO. PMID- 11869082 TI - Lipofection of rabbit chondrocytes and long lasting expression of a lacZ reporter system in alginate beads. AB - OBJECTIVE: Our aim was to investigate the maintenance of the transfection status of non-viral transfected chondrocytes in an alginate culture system. DESIGN: Chondrocytes harvested from rabbit knees were isolated by sequential digestion and cultivated in monolayer culture. At 60-70% cell density, chondrocytes were transfected with different transfection systems (FuGENE6, CaCl2, Lipofectin). A lac Z expression vector (pcDNA 3.1/Myc-His+ lacZ) was used as a reporter system. In order to improve transfection rates, hyaluronidase (4 U/ml) was used prior and during the transfection procedure. Thereafter, transfected cells were either kept in monolayer culture or embedded in alginate beads and kept in culture for up to the next 30 weeks. RESULTS: Transfection efficiency was maximal using FuGENE6TM/DNA at a ratio of 3:2 and hyaluronidase (4 U/ml). Transfection efficiency reached up to 40.8% (+/- 3.2%) after 36 h. In alginate beads lac Z positive cells declined to 8.5% +/- 3.3% after 4 weeks and to 4.6% +/- 3.2% after 12 weeks of culturing. After 30 weeks 3% of chondrocytes still expressed lac Z. In contrast, during culturing in monolayer, no lac Z expression was detectable after 4 weeks. Differentiation status of the chondrocytes was confirmed by histology and immunohistochemistry methods. CONCLUSIONS: After successful gene transfer to rabbit chondrocytes the alginate system made it possible to culture lipofected chondrocytes phenotypically stable. Genetically engineered chondrocytes express the lac Z reporter gene over a period of at least 30 weeks. This transfection and culture system provides a promising tool to further investigate the over-expression of growth factors and enzyme inhibitors. PMID- 11869083 TI - Integrative articular cartilage repair: dependence on developmental stage and collagen metabolism. AB - OBJECTIVES: The objectives of this research were to determine whether the integrative repair of bovine cartilage explants was dependent on developmental stage, and whether observed differences in integration with developmental stage were related to deposition of newly synthesized collagen and lysyl oxidase mediated collagen cross-linking. METHODS: Pairs of fetal, newborn calf, and adult bovine cartilage blocks were cultured in partial apposition for 2 weeks in medium supplemented with serum, ascorbate, and [3H]proline. Following culture, mechanical integration between apposed cartilage blocks was assessed by measuring adhesive strength in a single-lap shear configuration. Formation and stabilization of newly synthesized protein and collagen was investigated by determination of [3H]proline and [3H]hydroxyproline in tissue digests and guanidine extracts. RESULTS: Calf cartilage exhibited a relatively high integrative repair phenotype, achieving an adhesive strength that was three--four fold that of adult or fetal specimens. The low and high integrative repair phenotypes appeared related in part to different levels of collagen biosynthesis, which was approximately four--five-fold higher in calf cartilage samples than in the adult. However, fetal cartilage also exhibited a high level of biosynthesis. The different integrative repair phenotypes were not associated with marked differences in the kinetics of chemical stabilization of newly synthesized collagen, as the proportion of incorporated [3H]proline and newly-formed [3H]hydroxyproline that was resistant to extraction by 4M guanidine-HCl following culture was similar for cartilage from all developmental stages. Integration of calf cartilage appeared to depend on lysyl oxidase-mediated collagen cross-link formation, since inclusion of beta-aminopropionitrile (BAPN) in the culture medium completely eliminated development of adhesive strength. BAPN treatment also increased the percentage of newly synthesized protein in the guanidine extracts from 10% to 36% of the total, and that of newly synthesized collagen from 2% to 20%, while having only slight inhibitory effects on overall protein and collagen biosynthesis. CONCLUSION: The finding that cartilage exhibits enhanced integrative repair at a certain developmental stage suggests that it may ultimately be possible to enhance repair when needed in clinical situations. PMID- 11869084 TI - The effect of bone remodeling inhibition by zoledronic acid in an animal model of cartilage matrix damage. AB - OBJECTIVE: The purpose of this work was to test the effect of inhibition of bone remodeling, through the use of the bisphosphonate, zoledronic acid, on cartilage matrix damage in an animal model of cartilage matrix damage. DESIGN: New Zealand white rabbits were divided into four groups for treatment purposes: (1) untreated controls; (2) injected into one knee joint with the cartilage matrix degradation enzyme, chymopapain; (3) injected into one knee joint with chymopapain and also given subcutaneous injections of the bisphosphonate, zoledronic acid, three times per week until sacrifice at either day 28 or 56 post-chymopapain-injection; (4) received only the zoledronic acid injections. At sacrifice, the knee joints were examined grossly and histologically, and biochemically for proteoglycan content. Urine samples were analysed, at intervals, for levels of collagen cross-links which are biochemical markers of cartilage and bone. RESULTS: Animals receiving both intraarticular chymopapain injections and subcutaneous zoledronic acid injections displayed a significantly lower degree of grossly and histologically detectable cartilage degeneration on the tibial articular surfaces (the articular surface displaying the greatest degree of degeneration) than did animals only receiving the chymopapain injections. In addition, urinary levels of collagen cross-links for bone and cartilage were significantly higher in those animals only receiving chymopapain injections. CONCLUSION: The bone resorption observed after chymopapain injection into the rabbit knee joint can be inhibited through the use of the bisphosphonate, zoledronic acid. Furthermore, zoledronic acid does not increase the level of cartilage degeneration and appears to provide some level of chondroprotection in this model. PMID- 11869085 TI - Local overexpression of adeno-viral IL-4 protects cartilage from metallo proteinase-induced destruction during immune complex-mediated arthritis by preventing activation of pro-MMPs. AB - OBJECTIVE: To determine whether IL-4 protects against metalloproteinase-induced cartilage destruction during immune complex mediated arthritis and to elucidate its mechanism. METHODS: Experimental immune complex arthritis (ICA) was raised by injecting lysozyme into the knee joints of mice which previously were given anti lysozyme antibodies. Three days before ICA induction, mice were injected into the right knee joint with either IL-4 expressing or empty control recombinant human type 5 adenovirus. Joint inflammation and cartilage destruction (PG depletion, erosion) was measured by histology of total knee joints. Aggrecan breakdown in cartilage caused by metalloproteinases (MMPs) was studied by immunolocalization using anti-VDIPEN antibodies. RESULTS: Four days after ICA induction, histological analysis showed comparable exudate and infiltrate in both groups. Depletion of proteoglycans as measured by loss of red staining was also comparable in both groups. IL-4 treatment inhibited MMP-mediated neoepitope expression by 90%. Moreover, cartilage matrix erosion was evident in all animals (10 out of 10 mice) in the control group and significantly diminished (only two out of ten mice) in the IL-4 treated group. Incubation of patellae with APMA, which activates latent MMPs resulted in VDIPEN expression which was not significantly different from control ICA indicating that comparable amounts of latent pro-MMPs are present in IL-4 treated arthritic knee joints. CONCLUSION: This study indicates that during ICA, IL-4 largely prevents MMP-mediated aggrecan breakdown and severe cartilage erosion. IL-4 does not inhibit production of latent MMPs by the chondrocyte but predominantly interferes with its activation. PMID- 11869087 TI - Identity of human trophoblast cell lines: lessons from ED(27). PMID- 11869088 TI - Comparative developmental anatomy of the murine and human definitive placentae. AB - The placenta of eutherian mammals is a remarkable biological structure. It is composed of both zygote-derived and maternal cells, and mediates the complex interactions between the mother and the fetus that are necessary for fetal growth and survival. While the genetic basis of human placental development and function is largely unknown, its understanding is of immense clinical importance because placentopathies of unknown genetic aetiology are thought to be the cause of many types of pregnancy complications including unexplained miscarriage and intrauterine growth retardation. The mouse is the best-studied mammalian experimental genetic model system and research is not restricted by the inherent ethical and practical limitations associated with the human. As a result, knowledge about the genetic control of mouse placental development has expanded greatly in recent years. In order for this to be of benefit to medical practice, extrapolations from murine to human placentation have to be made. However, comprehensive comparisons of the placentae of these two species are rare. This review therefore compares the developmental anatomy of the placenta between humans and mice with emphasis on structures and cell types that might be analogous between the two species. This could be of particular benefit to mouse developmental geneticists who study placental development and have an interest in the possible clinical implications of their work. PMID- 11869089 TI - Current understandings of the molecular genetics of gestational trophoblastic diseases. AB - Gestational trophoblastic disease (GTD) is a heterogeneous group of diseases characterized by abnormally proliferating trophoblastic tissues. This includes partial and complete hydatidiform moles, invasive mole, choriocarcinoma and placental site trophoblastic tumour. Cytogenetic studies revealed that hydatidiform moles contain either solely (as in complete moles) or an excess (as in partial moles) of paternal contribution to the genome. Genomic imprinting is believed to play a pivotal role in the pathogenesis of hydatidiform moles. However its precise role and mechanism remains poorly understood. Hydatidiform mole carries a potential of malignant transformation. Similar to other human cancers, malignant transformation in gestational trophoblastic tumours is likely a multistep process and involves multiple genetic alterations including activation of oncogenes and inactivation of tumour suppressor genes. In addition, expression of telomerase activity, altered expression of cell--cell adhesion molecules and abnormal expression of matrix metalloproteinases have also been reported in GTD. These represent disruption of the delicate balance and regulation of cellular processes including proliferation, differentiation, apoptosis and invasion. The significance of these alterations in the pathogenesis and malignant transformation of gestational trophoblastic diseases is reviewed in this paper. PMID- 11869090 TI - ED(27) trophoblast-like cells isolated from first-trimester chorionic villi are genetically identical to HeLa cells yet exhibit a distinct phenotype. AB - ED(27) trophoblast-like cells were prepared from human chorionic villus samples obtained at 9 weeks gestation and have been grown continuously in vitro without phenotypic drift for nearly a decade. These cells express many trophoblast markers, including cytokeratin, placental alkaline phosphatase (PLAP), secretion of 17beta-estradiol, and a microvillous apical surface. The ED(27) cell line is a useful model system for studies of placental cell biology and has been distributed to laboratories world-wide. However, experiments to investigate their relationship to primary villous cytotrophoblast have shown that these cells do not secrete detectable amounts of human chorionic gonadotropin in culture and, when digested with trypsin, disperse into individual cells. Furthermore, immunocytochemical studies demonstrated that, unlike villous cytotrophoblasts, ED(27) cells were immunoreactive with monoclonal antibodies recognizing some HLA Class I antigens. This was not HLA-G, however, as would be expected if these cells originated from extravillous cytotrophoblasts, but rather classical HLA-A, B which is thought not to be expressed by any trophoblast subpopulations. These inconsistencies prompted us to question the authenticity of the continuous cell line as it now exists. Genetic haplotype analysis using the polymerase chain reaction (PCR) revealed that ED(27) was genetically identically to the HeLa cell line. Inasmuch as HeLa cells have never been grown in the laboratory (DAK), the only possible origin of HeLa cell contamination of ED(27) cells was the WISH cell line, and further PCR analysis revealed that this cell line was also genetically identical to HeLa. Like ED(27) cells, HeLa cells and WISH cells synthesized small amounts of estrogen and were found to express PLAP and antigens recognized by the monoclonal antibodies ED822, directed against the syncytiotrophoblast, and J1B5 directed against villous cytotrophoblast. These results point out the need for adherence to rigorous and consistent quality control measures to assure the authenticity of cell lines used as in vitro model systems. PMID- 11869091 TI - Transforming growth factor beta expression in human placenta and placental bed during early pregnancy. AB - Normal human pregnancy depends on physiological transformation of spiral arteries. Pre-eclampsia and fetal growth restriction are associated with impaired trophoblast invasion and spiral artery transformation. Recent data obtained from studies on placenta suggest that temporal changes in expression of TGF-beta3 play a key role in trophoblast invasion and that over-expression of TGF-beta3 in pre eclampsia is responsible for inadequate trophoblast invasion. There are, however, no studies of specific TGF-betas in the placental bed throughout pregnancy although this is where the invasive trophoblast and spiral arteries are located. In this study we have used immunohistochemistry, Western blot analysis, ELISA and RT-PCR to examine the expression of TGF-beta1, TGF-beta2 and TGF-beta3 in placental bed biopsies and placentas from 7--19 weeks' gestation. The results show that TGF-beta1, 2 and 3 are expressed in the placenta throughout this time but the striking temporal changes in TGF-beta3 expression previously reported were not observed. Extravillous trophoblast within the placental bed expressed TGF-beta2 but not TGF-beta1 or TGF-beta3 while extracellular TGF-beta1 and cytoplasmic TGF-beta2 were detected in decidua. These data suggest that TGF-beta1 and TGF-beta2 but not TGF-beta3 may play a role in trophoblast invasion. PMID- 11869092 TI - Differential activity of cathepsin L in human placenta at two different stages of gestation. AB - The implantation of blastocyst depends on the invasiveness of the syncytiotrophoblast, which penetrates the maternal decidua to establish the placenta. Cathepsin L, a lysosomal cysteine protease over-expressed in a variety of human malignancies, has been implicated in tumour invasion and metastasis. Specific inhibitors of cathepsin L inhibit the invasion of amnion by murine tumour cells. Previous studies have revealed that tumour and trophoblast invasiveness are mediated by shared factors, like metalloproteinases and laminin. Several proteases closely related to cathepsin L have recently been reported in the placentae of different species. In the present study, we demonstrate the expression of cathepsin L in human placenta by immunohistochemical analysis and RT-PCR followed by Southern hybridization. The activity of cathepsin L against the synthetic dipeptidyl substrate, Cbz-Phe-Arg-N-Methylcoumarin, was assayed. E 64, a specific inhibitor of cathepsin L was used to confirm that the enzyme activity being measured was due to cathepsin L. We observed the specific activity of cathepsin L in first trimester placenta to be significantly higher as compared to the term placenta. However, the levels of placental cathepsin L mRNA were comparable at these two stages of gestation. The increased enzymatic activity of cathepsin L in the invasive phase of placentation taken together with its previously established role in tumour invasion and metastasis indicates the involvement of this protease in trophoblast invasion. PMID- 11869093 TI - Ectodomain shedding of cystinyl aminopeptidase from human placental membranes. AB - Cystinyl aminopeptidase (CAP; EC 3.4.11.3) is an integral protein of the placental membrane that is also found in a soluble form in maternal serum during pregnancy. CAP was found to be shed from human placental membranes in a temperature- and time-dependent process. The released form of CAP was hydrophilic as assessed by phase separation in Triton X-114 and high-speed centrifugation. This ectodomain shedding of CAP was inhibited by the hydroxamic acid-based compounds marimastat and BB3103. The inhibition profile for the shedding of CAP was distinct to that for the release of angiotensin converting enzyme, implicating the involvement of distinct zinc metallosecretases in the shedding of these two proteins. These results have implications for our understanding of the mechanism underlying the reduction in serum levels of CAP observed in certain pregnancy-related disorders, such as pre-eclampsia. PMID- 11869094 TI - Implantation in the nine-banded armadillo: how does a single blastocyst form four embryos? AB - In the course of a study on reproduction in the nine-banded armadillo, conceptuses between the beginning of implantation and primitive streak formation were examined to determine the manner of trophoblast differentiation during invasion of the endometrium and the sequence involved in formation of four identical quadruplets. The armadillo blastocyst implants in the fundic recess of the uterus. A single amnion and cup-shaped epiblastic plate are formed, and an exocelom develops between the amnion and trophoblast of the implantation site. Loss of the abembryonic trophoblast exposes both visceral and parietal endoderm to the uterine lumen, inverting the yolk sac. Continued expansion of the exocelom facilitates the intrusion of the forming conceptus into the uterine lumen and is accompanied by enlargement of the epiblastic plate. Separate areas of condensations of epiblast cells are the first indication of formation of the four identical quadruplets. The single layer of microvillous trophoblast with basal infoldings (designated absorptive trophoblast) is most likely to contribute extensively to movement of fluid into the exocelom. The resulting expansion of the exocelom not only enlarges the implantation site but also displaces the collapsing common amnion, limiting the amnion to the areas of the forming embryos. PMID- 11869096 TI - Non-invasive method of evaluation of trophoblast invasion of spiral arteries in monochorionic twins with discordant birthweight. AB - To test the hypothesis that variation in birthweight between twin pairs is due to discordant placental development, we determined spiral arterial blood flow by colour pulsed Doppler ultrasound scan. We prospectively studied 24 twin pregnancies in the late second trimester with (n=12) and without (n=12) inter pair difference in estimated birthweight of > or = 20 per cent. In the discordant growth group, there were seven cases with chronic twin-twin transfusion syndrome (TTTS) and five without. The blood flow in spiral artery of each twin's portion of the placenta was assessed by resistance index (RI) by colour flow pulsed Doppler within a 5 cm radius of cord insertion. In twins with discordant weight, RI was increased in the growth restricted (FGR) twin than the appropriate for gestational age (AGA) co-twin (0.46 +/- 0.02 vs 0.3 +/- 0.01; P< 0.001) and the control group (P< 0.001). However, delta RI was comparable between twins with and without TTTS (0.13 +/- 0.01 vs 0.19 +/- 0.02; P=NS). No such differences were found between concordant twin pairs (0.28 +/- 0.01 vs 0.29 +/- 0.1; P=NS) and AGA twins of the discordant growth group. This study indicates that growth restricted twins have increased resistance to blood flow in the spiral arteries than the AGA co-twins. This observation, therefore, suggests non-physiological remodelling of the maternal spiral arteries in response to migrating trophoblast in placental bed of FGR MC twins. PMID- 11869097 TI - Isolation and spontaneous immortalization of trophoblast-like ED (27) cell lines. PMID- 11869095 TI - Placental nutrient transfer capacity and fetal growth. AB - The objective of this study was to determine whether the ability of the human placenta to transfer glucose and fatty acids is related to normal fetal growth. The intrinsic nutrient transport capacity of the placenta was measured under standardized conditions during in vitro perfusion of 30 human term placentas and related to birth weight (range 2640-4640g), birth weight centile (8th-99th), ponderal index (2.43-3.69), placental weight (418-1030g) and placental:fetal weight (0.14-0.31). There was no statistically significant change in the rate of nutrient transfer per placenta or per kg fetal weight, with birth weight, birth weight centile, ponderal index, placental weight and placental:fetal weight. There was a weak but significant relationship (P=0.020, r(2)=9 per cent) between the ratio of glucose to fatty acid transport and birth weight centile, largely due to the high ratio found in the lowest birth weight quartile where the babies are thinnest. This study provides no evidence that placental nutrient transport capacity limits fetal growth across a wide range of birth weights in normal pregnancies. It is proposed that the fetus itself may regulate placental nutrient transport in vivo via the fetal cardiac output and the rate of fetal nutrient utilization. PMID- 11869099 TI - Stereoselective ring expansion via bicyclooxonium ion. A novel approach to oxocanes. AB - [reaction: see text] The stereoselective 1,4-rearrangement-ring expansion of tetrahydrofurans via bicyclo[3.3.0]oxonium ions was developed to synthesize oxocanes. On the basis of this rearrangement, the stereoselective synthesis of 2,8-syn-2,8-dimethyloxocane was accomplished. PMID- 11869100 TI - Dendrimer with rotaxane-like mechanical branching. AB - [reaction: see text] A dendrimer wherein the branching points are mechanical in nature has been synthesized. It contains two identical covalently linked bis dendrons and a core unit fused to two rings that encircle the two bis-dendrons. A "threading-followed-by-stoppering" approach is used in the template-directed synthesis of a precursor bis[2]rotaxane, which undergoes stopper exchange four times to yield the dendrimer in which the two bis-dendrons act as stoppers within the two [2]rotaxane subunits. PMID- 11869098 TI - Akanthomycin, a new antibiotic pyridone from the entomopathogenic fungus Akanthomyces gracilis. AB - [structure: see text] Organic extracts of the entomopathogenic fungus Akanthomyces gracilis ARS 2910 contained antibiotics active against Staphylococcus aureus. Bioassay-guided fractionation of the CH2Cl2 extract yielded the antibacterial compound akanthomycin as a mixture of atropisomers along with the closely related compounds 8-methylpyridoxatin and cordypyridone C. Akanthomycin was characterized using X-ray crystallography and NMR. PMID- 11869101 TI - Optically active cyclic hexapeptides with covalently attached pyrene probes: selective alkaline earth metal ion recognition using excimer emission. AB - [structure: see text] The synthesis of two optically active pyrene-modified cyclic hexapetides and their selectivity toward the complexation of alkaline earth metal ions are reported. Complexation was studied by optical and chiroptical methods. The cyclic peptides are forming 2:1 sandwich complexes with the metal ions. PMID- 11869102 TI - Enantiospecific, stereospecific total synthesis of (+)-majvinine, (+)-10 methoxyaffinisine, and (+)-N(a)-methylsarpagine as well as the total synthesis of the Alstonia bisindole macralstonidine. AB - [structure: see text] The enantiospecific stereospecific total synthesis of majvinine 1a, 10-methoxyaffinisine 1b, and N(a)-methylsarpagine 1c are reported; this method has also resulted in the total synthesis of the Alstonia bisindole macralstonidine 2. PMID- 11869103 TI - Negishi coupling between alpha-alkyl(aryl)thio vinyl zinc chloride and alpha bromo vinyl ether: a convergent synthesis of 2-alkoxy-3-alkyl(aryl)thiobuta-1,3 dienes. AB - [reaction: see text] A convergent approach for the stereoselective synthesis of 2 alkoxy-3-alkyl(aryl)thiobuta-1,3-dienes has been developed. It was found that Negishi coupling between alpha-alkyl(aryl)thio vinyl zinc chloride and alpha bromo vinyl ether or Negishi coupling between alpha-bromo vinyl sulfide and alpha alkoxy vinyl zinc chloride provided the best yield and stereoselectivity. PMID- 11869105 TI - Regioselective synthesis of indoles via reductive annulation of nitrosoaromatics with alkynes. AB - [reaction: see text] Indoles are produced regioselectively and in moderate yields by two new processes: (a) from the [CpRu(CO)2]2-catalyzed reaction of nitrosoaromatics (ArNO) with alkynes under carbon monoxide and (b) in a two-step sequence involving the (uncatalyzed) reaction of ArNO with alkynes, followed by reduction of the intermediate adduct. PMID- 11869104 TI - Asymmetric synthesis of the highly methylated tryptophan portion of the hemiasterlin tripeptides. AB - [reaction: see text] The asymmetric synthesis of the methylated tryptophan portion of hemiasterlin peptides is described. The key reactions are a SnCl4 mediated ring opening of epoxynitriles or epoxysulfones by N-methylindole followed by an asymmetric Strecker reaction. A second approach involving opening of glycidic esters by indoles is also described. PMID- 11869107 TI - Optically active 1,2-bis(1-arylhydroxymethyl) ferrocene: a new, efficient chiral ligand for scandium-catalyzed asymmetric Diels-Alder reaction. AB - [reaction: see text] The Sc(OTf)3/FERRODIOL (2) complex was prepared at -78 degrees C in CH2Cl2 in the presence of 2,6-lutidine and MS 4A. The chiral scandium Lewis acid-catalyzed asymmetric Diels-Alder reaction of cyclopentadiene (3) with 3-acyloxazolidin-2-ones (4) effectively produced the adduct (5) in a high yield with good selectivity, i.e., endo/exo = 90:10 up to 91% ee (endo). PMID- 11869106 TI - A highly efficient, selective, and general method for the synthesis of conjugated (all-E)-oligoenes of the (CH=CH)n type via iterative hydrozirconation-palladium catalyzed cross-coupling. AB - [reaction: see text] A linear iterative method for the construction of (all-E) oligoenes of the (CH=CH)n type via hydrozirconation-palladium-catalyzed cross coupling with (E)-1-bromo-4-trimethylsilyl-1-buten-3-yne is described. This method promises to provide an efficient, selective, and general route to oligoene macrolide antibiotics and other related natural products. PMID- 11869108 TI - Total synthesis of the cyclic heptapeptide Argyrin B: a new potent inhibitor of T cell independent antibody formation. AB - [structure: see text] The total synthesis of Argyrin B (1) is presented using a synthetic plan that is convergent and flexible and conserves the stereogenic centers. The unusual amino acid 4-methoxy tryptophan (6) was obtained via an enzymatic resolution. Cyclization followed by oxidative elimination of the phenylseleno cysteine to the sensitive dehydroalanine afforded synthetic 1. PMID- 11869109 TI - Total synthesis of (+/-)-jamtine using a thionium/N-acyliminium ion cascade. AB - [reaction: see text] The first total synthesis of (+/-)-jamtine (4), a tetrahydroisoquinoline alkaloid reputed for its therapeutic properties, is described. The key step involves a tandem thionium/N-acyliminium ion cyclization from enamido sulfoxide 13. The cascade process takes place with high diastereoselectivity and in excellent yield. PMID- 11869110 TI - Synthesis and photophysical properties of new conjugated fluorophores designed for two-photon-excited fluorescence. AB - [structure: see text] Novel elongated push-push fluorophores (e.g., 9) were synthesized by 2-fold Sonogashira or Wittigminus signHorner reactions. Modulation of the length and topology of the conjugated connectors allows tuning of their photophysical properties. In addition, their photoluminescence can be adjusted by playing on polarity. Derivatives combining enhanced two-photon absorption cross section (sigma2) in the visible red and high fluorescence quantum yield (Phi) have been obtained. Such fluorophores hold promise for nonlinear imaging of biological systems. PMID- 11869111 TI - A chiral 28-membered macrocycle with symmetry and structure similar to that of trans-cyclooctene. AB - [reaction: see text] A bridged N,N-di(aryl)-1,2,4,5-benzenediimide was synthesized in which restricted rotation led to two diasteriomeric conformations at room temperature. The more stable syn-macrocycle is achiral, whereas the strained anti-macrocycle possesses planar chirality similar to that of trans cyclooctene. The structure was characterized by X-ray crystallography, and the enantiomers were resolved by chiral chromatography. PMID- 11869112 TI - Electron-poor benzonitriles as labile, stabilizing ligands in asymmetric catalysis. AB - [structure: see text] A chiral palladium catalyst [(S) MeObiphep)Pd(NCAr)2(SbF6)2, (S)-4c], has been developed for a variety of asymmetric transformations. (S)-4c is bench-stable and has activity comparable to that of the nitrile free Lewis acid catalyst for Diels-Alder, hetero-Diels-Alder, and glyoxylate-ene reactions. PMID- 11869113 TI - Toward novel polyacetals by transacetalation techniques: dendrimeric diacetals. AB - [structure: see text] A new approach to polyacetal systems using sequential transacetalation and protection-deprotection techniques was developed for the preparation of macromolecular polyacetals and applied to secure new dendrimers with 2,4,8,10-tetraoxaspiro[5,5]undecane dendrons from pentaerythritol and polyaldehydes. These novel dendrimers, featuring a 1,3,5-benzene-tricarbaldehyde core, viz., the dodecaol (18) and its hexaacetal (19), were prepared, and molecular modeling revealed peculiar dendron convergent structures above the core, due to intramolecular pi-stacking interactions, reinforced by H-bonding of multifunctional termini. PMID- 11869114 TI - Aromaticity of butalene and its homologues. AB - [structure: see text] The aromaticity of a series of fused cyclobutadiene systems is discussed in terms of their resonance energies. While there is considerable variation in their resonance energies per pi electron, all members of the series are calculated to be antiaromatic, though to a lesser degree than the parent cyclobutadiene. PMID- 11869115 TI - Soluble polymer-supported catalysts containing azo dyes. AB - [structure: see text] Azo benzene derivatives were incorporated into soluble polymer-bound catalysts by two different approaches. The first was to attach the dye to the polymer-bound catalysts, using the dye as an innocent spectator to study the phase preference, concentration, and recoverability of a catalyst. The second approach used an azo dye as a ligand to form an effective soluble polymer bound Pd(II) catalyst for Heck reactions. PMID- 11869116 TI - Solid-phase synthesis of novel isoxazolocyclobutanones and isoxazolinocyclobutenones. AB - [reaction: see text] The preparation of novel isoxazolocyclobutanone and isoxazolinocyclobutenone derivatives via a traceless solid-phase sulfone linker strategy is described. Key steps in the solid-phase protocol reported here include (i) sulfinate right arrow sulfone alkylation, (ii) four-member ring formation by sulfone dianion alkylation, (iii) heterocycle formation by nitrile oxide 1,3-dipolar cycloaddition, and (iv) traceless product release by cyclobutanol right arrow cyclobutanone oxidation with concomitant linker cleavage by sulfinate elimination. PMID- 11869117 TI - Synthesis of monodisperse oligo-p-phenylenes via rhodium-catalyzed alkyne cyclotrimerization. AB - [structure: see text] A novel synthesis of monodisperse oligo-p-phenylenes is described that features alkyne cyclotrimerization of ether-linked p dialkynylarenes. This route introduces solubilizing dihydrofuran and alcohol functional groups that may improve processing of the corresponding polymeric p phenylene materials. PMID- 11869118 TI - Stereoselective synthesis of vancosamine and saccharosamine glycals via tungsten catalyzed alkynol cycloisomerization. AB - [reaction: see text] A stereoselective synthesis of the C-3 branched amino glycals of vancosamine and saccharosamine is described that features a tungsten carbonyl catalyzed cycloisomerization of the corresponding alkynyl alcohol. PMID- 11869119 TI - Tandem catalysis and self-assembly: a one-pot approach to functionalized polymers. AB - [reaction: see text] Side-chain functionalized polymers possessing terminal palladated SCS pincer complexes at each repeat unit were synthesized via ring opening metathesis polymerization. These palladium centers function as both well defined Heck catalysts and recognition units capable of quantitative self assembly of pyridine-containing molecules. Exploitation of both the catalytic and self-assembly properties has led to the development of a controlled, one-pot tandem catalysis/self-assembly sequence for the synthesis of functionalized polymers. PMID- 11869120 TI - First synthesis of 10 alpha-(trifluoromethyl)deoxoartemisinin. AB - [reaction: see text] A novel, nonacetal (trifluoromethyl)deoxoartemisinin was prepared with good stereoselectivity. This compound was obtained by debromination of the 10 alpha-CF3-10-bromodeoxoartemisinin in the presence of tributyltin hydride at reflux in toluene without alteration of the endoperoxide bridge. It presented a reasonable antimalarial activity. PMID- 11869121 TI - Catalytic hydrophosphination of styrenes. AB - [reaction: see text] The first example of intermolecular hydrophosphination of styrenes catalyzed by Ni and Pd complexes is described. The reaction of Ph2PH with styrene, 4-vinylpyridine, 2-vinylpyridine, 4-methoxystyrene, 2 methoxystyrene, and 5-vinyl-2-methylpyridine in benzene under Ni[P(OEt)3]4 catalysis proceeds with high yield and selectivity to give only anti-Markovnikov product. PMID- 11869122 TI - Synthesis of novel alpha-substituted and alpha,alpha-disubstituted amino acids by rearrangement of ammonium ylides generated from metal carbenoids. AB - [reaction: see text] A new and general four-step synthesis of protected alpha substituted and alpha,alpha-disubstituted amino acids has been developed. The key step involves intramolecular ammonium ylide generation from a copper carbenoid with concomitant [2,3] rearrangement. The aromatic template serves as a tether, protecting group, and activating group for peptide coupling. The ylide rearrangement products can be converted into protected cyclic amino acids by ring closing metathesis. PMID- 11869123 TI - Fluoroalkyl alpha,beta-unsaturated imines. Valuable synthetic intermediates from primary fluorinated enamine phosphonates. AB - [reaction: see text] A simple method for the preparation of fluoroalkyl allylamines or alpha,beta-unsaturated ketones by an olefination reaction of primary enamine phosphonates and aldehydes, followed by selective reduction with hydrides or hydrolysis, is reported. Fluorinated beta-amino nitriles are also obtained by an olefination reaction of primary enamine phosphonates with aldehydes and subsequent addition of metalated acetonitrile. PMID- 11869124 TI - Reaction of 1,4-phthalazinedione with furfural: formation of the [5,6]benza-3a,7a diazaindane system via an unusual skeletal rearrangement. AB - [reaction: see text] Oxidation of phthalahydrazide (1) with lead tetraacetate in the presence of furfural (3a) in methylene chloride gives [5,6]benza-3a,7a-diaza 3-carboxylindane-4,7-dione-1-ene (7a) in 64% yield. 5-Methylfurfural (3b) also reacted similarly to give the product 7b in 46% yield. Reaction of phathalazine 1,4-dione with thiophene-2-carboxaldehyde gives N-(2 formylthiophene)phthalahydrazide. PMID- 11869126 TI - Microwave-assisted ketone-ketone rearrangement: an improved synthesis of 3-(4 alkoxyphenyl)-3- methylbutan-2-ones. AB - [reaction: see text] A novel procedure for the preparation of 3-(4-alkoxyphenyl) 3-methylbutan-2-one in excellent yield is described via polymer-supported AlCl3 catalyzed rearrangement of 1-(4-ethoxyphenyl)-2,2-dimehtylpropan-1-one, followed by O-alkylation under microwave irradiation condition. PMID- 11869125 TI - Palladium-catalyzed synthesis of trans-4-(N,N-bis(2-pyridyl)amino)stilbene. A new intrinsic fluoroionophore for transition metal ions. AB - [reaction: see text] The title compound (1) has been synthesized via sequential Pd-catalyzed amination reactions and investigated as an intrinsic fluoroionophore. The efficiency in the synthesis of 1 strongly depends on the order of couplings among the substrates. Compound 1 displays fluorescence quenching upon the binding of transition metal ions, where the binding-triggered conformational twisting and in turn the inhibition of internal charge transfer (ICT) play an important role. PMID- 11869128 TI - Dearomatizing annelation of five-membered rings to naphthalenes by organolithium cyclization. AB - [reaction: see text] gamma-Lithiopropylnaphthalenes and their oxa- and aza tethered analogues cyclize by nucleophilic addition of the organolithium to the naphthalene ring. The resulting benzyllithiums react stereoselectively with electrophiles to give dearomatized tricyclic products with structural similarity to the arylnaphthalene lignans. PMID- 11869127 TI - Spongistatin synthetic studies. An efficient, second-generation construction of an advanced ABCD intermediate. AB - [reaction: see text] A short, efficient, and stereocontrolled synthesis of (-)-4, an advanced ABCD subunit of the spongistatins, has been achieved. Central to the synthetic strategy is the multicomponent linchpin union of silyl dithianes with epoxides to access both the AB and CD fragments. Fragment coupling was then achieved via an efficient stereoselective aldol reaction. The linear sequence required 22 steps and proceeded in 4.0% overall yield. PMID- 11869129 TI - Comparative study of anionic and radical cyclization for the preparation of 1,3 dimethylindans: highly stereoselective preparation of cis-1,3-disubstituted indans via intramolecular carbolithiation. AB - [reaction: see text] The preparation of 1,3-dimethylindans from 4-(2-bromophenyl) 1-pentene (1) and 2-(2-iodo-1-methylethyl)styrene (2) substrates via radical mediated cyclization and intramolecular carbolithiation has been investigated. Although cyclization of the radical derived from either substrate proceeds with modest selectivity for the cis-isomer, as does cycloisomerization of the aryllithium derived from substrate 1 (cis/trans approximately 2), intramolecular cyclization of the alkyllithium derived from substrate 2 is a highly cis selective process (cis/trans = 12). PMID- 11869130 TI - Novel C3V-symmetric tripodal scaffold, triethyl cis,cis,cis-2,5,8- tribenzyltrindane-2,5,8-tricarboxylate, for the construction of artificial receptors. AB - [reaction: see text] A novel C3V-symmetric scaffold, trindane 7, has been efficiently synthesized from 1,3,5-tris(bromomethyl)-2,4,6 tris(chloromethyl)benzene (1) in six steps with 47% overall yield. The control of all-syn stereochemistry in the tribenzylation step has been achieved by blocking one side of the trindane ring as metal carbonyl complexes. The potential utility of trindane 7 as a receptor skeleton has been examined with a urea derivative 12 toward several anionic guests. PMID- 11869131 TI - Synthetic studies on the ingenane diterpenes. An improved entry into a trans intrabridgehead system. AB - [reaction: see text] The efficient construction of an ingenol intermediate exhibiting "insideminus signoutside" intrabridgehead stereochemistry is reported. The sequence features the net conversion of a cis-intrabridgehead compound into a highly strained trans-species via palladium-mediated isomerization of an allylic epoxide followed by a low-temperature alkoxide-accelerated 1,5-hydrogen migration. PMID- 11869132 TI - Synthesis of cyclic dienamide using ruthenium-catalyzed ring-closing metathesis of ene-ynamide. AB - [reaction: see text] Ring-closing metathesis of ene-ynamide using the second generation Grubbs' catalyst produced nitrogen-containing heterocycles, which have dienamide moieties, in high yields. Diels-Alder reaction of the cyclized product and dienophile proceeded smoothly to afford a bi- or tricyclic compound. PMID- 11869133 TI - Ni-catalyzed highly regio- and chemoselective cocycloaddition of nonconjugated diynes with 1,3-diynes: a novel method for polysubstituted arylalkynes. AB - [reaction: see text] Effective nickel-catalyzed cocycloaddition of nonconjugated diynes with conjugated diynes yields polysubstituted arylalkynes in good to excellent yields with high regio- and chemoselectivity. PMID- 11869134 TI - Barbituric acid initiated rearrangement of 2,2'-pyridil into 5,5'-(2 pyrilidine)bisbarbituric acid. AB - [reaction: see text] In the study of the barbituric acid initiated 2,2'-pyridil rearrangement, a very efficient synthetic procedure (isolated yield 80-90%) for the preparation of useful 2-pyrilidenes 3 was developed. PMID- 11869135 TI - Rapid route to 3,4-substituted indoles via a directed ortho metalation-retro Mannich sequence. AB - [reaction: see text] In the presence of NXS (X = Br, I, Cl), gramine derivatives 1, derived by combined directed ortho metalation (DoM)-cross-coupling sequences, rapidly undergo retro-Mannich fragmentation (2) to afford 3-halo indoles 3 in 37 88% yields. A conceptually new methodology to diverse 3,4-substituted indoles (10, 11, 13) is thereby introduced. PMID- 11869137 TI - First alkyl radical additions to (eta(6)-arene)tricarbonylmanganese complexes. AB - [reaction: see text] Reactions of alkylmercury chlorides with arene manganese tricarbonyl complexes in the presence of NaI led to the formation of the addition reduction products. The mechanism was postulated to be the alkyl radical addition to ArMn(CO)3+ cation to form the corresponding 17 valence electron intermediate, which was then reduced by alkylmercury chloride via a singlet electron transfer process to afford the product and regenerate an alkyl radical. PMID- 11869136 TI - Synthesis of (-)-ilimaquinone via a radical decarboxylation and quinone addition reaction. AB - [reaction: see text] A stereoselective synthesis of (-)-ilimaquinone (4) is presented. The synthetic strategy is based on a novel radical decarboxylation and quinone addition methodology that produces quinone 7 from reaction of thiohydroxamic acid derivative 8 with benzoquinone (9). Final functionalization of 7 to ilimaquinone (4) is achieved by exploring the electronic effects of the residual thiopyridyl group. PMID- 11869138 TI - New methodologies for the oxidation of Fischer carbene complexes: synthesis of hydrazides. AB - [reaction: see text] We report new, high-yield methodologies for oxidizing Fischer carbenes, particularly hydrazinocarbene complexes. The reagents traditionally used to oxidize Fischer carbenes have failed because of the stability of hydrazinocarbene complexes and the easy oxidation of formed hydrazides in the reaction conditions. The three newly developed methodologies are very mild, fast, efficient, and complementary. Differently functionalized hydrazinocarbene complexes can be oxidized to afford new hydrazides. PMID- 11869139 TI - Kinetic separation methodology for the stereoselective synthesis of (E)- and (Z) alpha-fluoro-alpha,beta-unsaturated esters via the palladium-catalyzed carboalkoxylation of 1-bromo-1-fluoroalkenes. AB - [reaction: see text] Methodology for the stereoselective preparation of both (E)- and (Z)-alpha-fluoro-alpha,beta-unsaturated esters is described. 1-Bromo-1 fluoroalkenes (E/Z approximate 1:1) can be isomerized to high E/Z ratio mixtures, which participate in palladium-catalyzed carboalkoxylation and lead to (Z)-alpha fluoro-alpha,beta-unsaturated esters in high stereoselectivity. The same starting material can also be kinetically reduced to get an E/Z ratio of 0:100; similar carboalkoxylation reaction at 70 degrees C affords (E)-alpha-fluoro-alpha,beta unsaturated esters stereospecifically. PMID- 11869140 TI - Asymmetric synthesis of beta-amino alcohols by reductive cross-coupling of benzylideneamine with planar chiral benzaldehydes. AB - [reaction: see text] Samarium iodide mediated reductive cross-coupling of N-tosyl benzylideneamine with benzaldehydes or the corresponding chromium complexes gave syn-beta-amino alcohol derivatives. A dynamic kinetic resolution of a configurationally equilibrated reactive species occurred in the cross-coupling with planar chiral benzaldehyde chromium complexes. PMID- 11869141 TI - Domino reactions of amidines with methyl 2-chloro-2-cyclopropylideneacetate as an efficient access to cyclobutene-annelated pyrimidinones. AB - [reaction: see text] An efficient one-step synthesis of 2,4 diazabicyclo[4.2.0]octa-1(6),2-dien-5-ones 3 from methyl 2-chloro-2 cyclopropylideneacetate (1) and amidines 2a-c as well as N,N-dimethylguanidine (2d) is described. Similar to the benzocyclobutenes, the cyclobutene-annelated pyrimidones 3 undergo thermal ring opening and the resulting o-quinodimethane analogues readily cycloadd dienophiles to yield tetrahydroquinazolone derivatives. PMID- 11869142 TI - Synthesis of thiazolines linked to a difluoromethylphosphonate diester via dithioester chemistry. AB - [reaction: see text] A two-step, high-yielding synthesis of delta(2)-thiazolines containing a difluoromethylphosphonate diester moiety has been devised using a building block approach. Racemic or chiral beta-amino alcohols and diols were coupled with methyl difluoro(diethoxyphosphono)dithioacetate to give predominantly the corresponding beta-hydroxythioamides, which were then cyclized to provide a series of novel substituted delta(2)-thiazolines. PMID- 11869143 TI - Metal trifluoromethanesulfonate-catalyzed regioselective borane-reductive ring opening of benzylidene acetals: a concise synthesis of 1,4-dideoxy-1,4-imino-L xylitol. AB - [reaction: see text] A highly regioselective borane-reductive ring opening of the 4,6-O-benzylidene-D-hexopyranosides to the corresponding 6-alcohols in excellent yields at room temperature via various metal trifluoromethanesulfonates as catalysts is described here. Its application in the synthesis of 1,4-dideoxy-1,4 imino-L-xylitol is also highlighted. PMID- 11869144 TI - An 8pi electron electrocyclization leading to a 9,19-methano-bridged analogue of 1 alpha,25-dihydroxyvitamin D3. AB - [reaction: see text] Lindlar semihydrogenation of a vitamin D type trienyne leads spontaneously to 9 alpha,19-methano-1 alpha,25-dihydroxyvitamin D3. The intermediate tetraene resulting from the reduction undergoes a rapid, stereoselective 8pi electron electrocyclization affording a novel steroid containing a linearly fused ABC (six-eight-six) 1,3,5-cyclooctatriene carbon framework. PMID- 11869145 TI - Bis(isoquinoline N-oxide) pincers as a new type of metal cation dual channel fluorosensor. AB - [structure: see text] A new type of donor-spacer-acceptor podand system has been synthesized and proved as an efficient dual channel fluorosensor for Li+, Mg2+, and Ca2+. The known ability for the N-oxide function to bind Lewis acids is the key step in the appearance of a new emitting charge-transfer (CT) excited state. The occurrence of this CT state for alkaline earth (Mg2+ and Ca2+) and not for alkaline metals (Li+) provided a new type of dual channel fluorosensors. PMID- 11869146 TI - Studies toward the total synthesis of garsubellin A: a concise synthesis of the 18-epi-tricyclic core. AB - [reaction: see text] During studies directed toward the total synthesis of garsubellin A, a concise stereocontrolled synthesis of the 18-epi-tricyclic compound 3 was achieved. Key steps were a one-pot stereoselective construction of the bicyclic lactone 23 followed by a formal migration to the bicyclo[3.3.1]nonane-1,3,5-trione and an intramolecular Wacker-type tetrahydrofuran ring formation. PMID- 11869147 TI - Amidoglycosylation via metal-catalyzed internal nitrogen atom delivery. AB - [reaction: see text] Rhodium and copper acyl nitrenoids are likely intermediates in amidoglycosylation reactions of allal 3-carbamates. Iodine(III)-mediated nitrenoid formation, interaction of this species with the glycal enol ether pi system, and highly beta-stereoselective glycosylation occur in a one-pot process that requires no additional Lewis acid activation. PMID- 11869149 TI - The face in the mirror. PMID- 11869150 TI - Voluntary upper-extremity movements in patients with unilateral peripheral vestibular hypofunction. AB - BACKGROUND AND PURPOSE: People with peripheral vestibular pathology demonstrate motor impairments when responding and adapting to postural platform perturbations and during performance of sit-to-stand and locomotor tasks. This study investigated the influence of unilateral peripheral vestibular hypofunction on voluntary arm movement. SUBJECTS AND METHODS: Subjects without known neurological impairments and subjects with vestibular impairments performed 3 voluntary arm movements: an overhead reach to a target, a sideward reach to a target, and a forward flexion movement through 90 degrees. Subjects performed these tasks under precued and choice reaction time conditions. During all tasks, body segment motion was measured. Head velocity measurements were calculated for the side task only. RESULTS: Subjects with vestibular loss restricted upper body segment motion within the frontal and transverse planes for the 90-degree and overhead tasks. Average angular head velocity was lower for the group with vestibular hypofunction. Task uncertainty (the introduction of a choice reaction time paradigm) differentially influenced the groups regarding head velocity at target acquisition. DISCUSSION AND CONCLUSION: Individuals with vestibular loss altered their performance of voluntary arm movements. Such alterations may have served to minimize the functional consequences of gaze instability. PMID- 11869151 TI - Three perspectives on physical therapist managerial work. AB - BACKGROUND AND PURPOSE: The nature of managerial work in the commercial sector has not been studied since the 1970s, and little is known about the work of managers in the health care sector. In this study, the perceived importance of managerial role and skill categories among 3 groups of physical therapists were studied to better understand the work priorities of physical therapist managers. SUBJECTS: Two groups of subjects were physical therapist managers in hospitals or private practices. A third group consisted of faculty members in professional physical therapist education programs. METHODS: Respondents (n=343) rated the importance of 75 managerial activities. Responses related to 16 predetermined work categories were placed in rank order by group. A multivariate analysis of variance (MANOVA) was used to identify differences among groups. RESULTS: All groups identified communication, financial control, entrepreneur, resource allocator, and leader as the 5 most important categories and rated technical expert and figurehead as least important. The MANOVA showed differences between faculty members and private practice managers in 15 work categories, between hospital-based managers and private practice managers in 9 categories, and between faculty members and hospital-based managers in 8 categories. DISCUSSION AND CONCLUSION: Work setting appears to have an impact on level of importance placed on managerial work categories. The strongest candidates for "universal" physical therapist managerial work categories were communication, financial control, and resource allocator. PMID- 11869152 TI - Physical therapists' perceptions of factors influencing the acquisition of motor abilities of children with cerebral palsy: implications for clinical reasoning. AB - BACKGROUND AND PURPOSE: Evidence supporting factors predicting motor change for children with cerebral palsy is minimal. A consensus exercise using focus groups and survey methods was conducted to identify factors perceived to affect the acquisition of basic motor abilities among children with cerebral palsy from the time of diagnosis to 7 years of age. SUBJECTS: Fifty-seven physical therapists participated in one of 12 focus groups, and 60 physical therapists participated in a follow-up questionnaire survey via mail. METHODS: The nominal group technique was used to conduct the focus groups. RESULTS: Participants reached consensus about 12 factors in 4 constructs, which we called: (1) primary impairments (muscle tone/movement patterns, distribution of involvement, balance, and sensory impairment), (2) secondary impairments (range of motion/joint alignment, force production, health, and endurance), (3) personality characteristics (motivation), and (4) family factors (support to child, family expectations, and support to family). DISCUSSION AND CONCLUSION: The recognition of potential determinants of motor change could assist in the clinical reasoning that physical therapists use when planning interventions for children with cerebral palsy. Participants identified a set of variables, some of which are found in the literature, that can provide foundation knowledge for decision making and research on factors that bring about change in motor ability among children with cerebral palsy. PMID- 11869153 TI - Relationships among lifting ability, grip force, and return to work. AB - BACKGROUND AND PURPOSE: The relationship between functional capacity evaluation (FCE) data and work disability has not been studied. The validity of FCE testing results in terms of subsequent return to work (RTW) was the focus of this exploratory study. SUBJECTS AND METHODS: Six hundred fifty adults of working age were evaluated as part of a standardized FCE. Clients were contacted by telephone 6 months after the FCE to determine their work status. Predictor variables were gender, age, time off work, maximum safe loads during 3 dynamic lifts, and isometric grip force. Other variables measured were whether or not the client returned to work (RTW-Y/N) and level of return to work (RTW level). RESULTS: A multivariate logistic regression analysis demonstrated that the more time a worker was away from work, the less likely was RTW. Male subjects were less likely to return to work than female subjects. The more weight lifted from floor to waist, the more likely was RTW. The logistic regression equation correctly classified 80.3% of the subjects who returned to work and 56.6% of the subjects who did not return to work. Each of the 3 lift tests was related to RTW level, whereas the grip force tests were not related to either RTW-Y/N or RTW level. DISCUSSION AND CONCLUSION: Time off work and gender were the strongest predictors of RTW, but certain FCE subtests of lifting were related to RTW and RTW level for people with work-related chronic symptoms. Grip force was not related to RTW. PMID- 11869154 TI - A calibration study of therapeutic ultrasound units. AB - BACKGROUND AND PURPOSE: Physiological effects of therapeutic ultrasound (US) are dependent on the intensity and duration of application. The purpose of this study was to test US machines used in clinical settings for proper calibration of time and power output. METHODS: Measurements of power output and timer accuracy were obtained from 83 US units in clinical use. The machines were tested at 4 intensity settings (0.5, 1.0, 1.5, and 2.0 W/cm2) using a continuous waveform and a 1-MHz frequency. The measured intensities were converted to percentages of error and compared with the +/-20% standard. RESULTS: Of the machines tested, 32 (39%) were outside the calibration standard for at least one output setting. Of these machines, 15 (18%) were above the +20% standard, and 17 (21%) were below the -20% standard for at least one output setting. Of the 32 machines outside the standard, 26 (31%) were outside the standard for 2 or more settings, and 3 (4%) produced no output at any of the settings. Of the mechanical timers tested, 7 (28%) were outside of the +/-10% standard for timer accuracy at the 5-minute interval, and 6 (24%) were outside of the standard at the 10-minute interval. All digital timers tested were within the standard. DISCUSSION AND CONCLUSION: More than one third of machines tested in this study were outside the standard for power output, and approximately one fourth of the mechanical timers were outside the standard. Therefore, further improvements in the accuracy of US machine calibration are needed. PMID- 11869155 TI - Fear of falling. PMID- 11869156 TI - Are physical agents the same as rehabilitation? PMID- 11869158 TI - More questions than answers. PMID- 11869157 TI - Is manual physical therapy distinct from physical therapy clinical practice? PMID- 11869159 TI - Sustainable undergraduate education and professional competency. PMID- 11869160 TI - Waking cervical pain and stiffness, headache, scapular or arm pain: gender and age effects. AB - A random population-based study using a structured telephone questionnaire was undertaken to determine the frequency, duration and prevalence of waking cervical spine pain and stiffness, headache, and aching between the scapulae or in the arm and their relationship to age and gender. Seventy-six per cent of households contacted completed the questionnaire, yielding a total of 812 questionnaires. Subjects most commonly reported waking with scapular or arm pain (27%) followed by headache (19%), cervical pain (18%) and cervical stiffness (17%). The majority of subjects reported that symptoms lasted for more than an hour on one or two occasions during the week. Subjects who reported the presence of one type of waking symptom were significantly more likely to report other waking symptoms. Females were significantly more likely to wake with a headache than males (OR 2.5, 95% CI 1.6 to 4.0), while all subjects aged over 60 years were significantly less likely to wake with a headache than subjects in other age groups (OR 0.6, 95% CI 0.4 to 1.0). Females exhibited a decline in waking cervical pain, stiffness and headache with increasing age. In contrast, males exhibited a peak prevalence of waking cervical pain, stiffness and headache in the 40 to 59 year age group. The prevalence of waking with aching between the scapulae or in the arm behaved differently from the other symptom groups in both genders. PMID- 11869162 TI - An evaluation of Superthumb and the Kneeshaw device as manual therapy tools. AB - This research evaluated two hand-held tools (Superthumb and Kneeshaw device) that have been developed in order to reduce hand pain associated with the performance of manual therapy. Two studies were conducted: one evaluated the ability to perceive elastic stiffness with the devices and the other evaluated physiotherapist and patient comfort when the devices were used to apply a mobilisation to the lumbar spine. In the first study we found that the two tools and the pisiform grip provided equivalent ability to detect small differences in elastic stiffness, however the tools introduced a bias so that the stiffness stimuli felt stiffer than when assessed with the pisiform grip. In the second study we found that the two tools were substantially less comfortable than the pisiform grip, for both patient and therapist, when a therapist applied a Grade III mobilisation to the lumbar spine. The results suggest that neither tool, in its current form, is suitable for clinical practice. PMID- 11869161 TI - Hand positioning sense in children with spina bifida myelomeningocele. AB - This study was undertaken to establish whether children with myelomeningocele have abnormal kinaesthesia of the hands. Twenty-one children with myelomeningocele and 21 control children aged between six and 12 years were involved in the study. The level of kinaesthetic awareness in the hands was measured by examining the child's ability to copy hand positions, using visual cueing and kinaesthetic cueing. Both accuracy and speed of copying hand gestures were assessed. Children with spina bifida were significantly less accurate in achieving hand positions than the control group (chi square(1) = 22.60, p < 0.001), with 73% of the children with spina bifida achieving accurate replications compared with 87% in the control group. Furthermore, children with myelomeningocele were shown to be slower than the controls (F(1,280) = 15.49, p < 0.001). The impaired kinaesthetic awareness found in this study is considered to be one of the factors behind the poor hand function observed in children with myelomeningocele. PMID- 11869163 TI - Measurement of functional ability following traumatic brain injury using the Clinical Outcomes Variable Scale: a reliability study. AB - This study determined the inter-tester and intra-tester reliability of physiotherapists measuring functional motor ability of traumatic brain injury clients using the Clinical Outcomes Variable Scale (COVS). To test inter-tester reliability, 14 physiotherapists scored the ability of 16 videotaped patients to execute the items that comprise the COVS. Intra-tester reliability was determined by four physiotherapists repeating their assessments after one week, and three months later. The intra-class correlation coefficients (ICC) were very high for both inter-tester reliability (ICC > 0.97 for total COVS scores, ICC > 0.93 for individual COVS items) and intra-tester reliability (ICC > 0 97). This study demonstrates that physiotherapists are reliable in the administration of the COVS. PMID- 11869164 TI - Evidence for physiotherapy practice: a survey of the Physiotherapy Evidence Database (PEDro). AB - Evidence-based practice involves the use of evidence from systematic reviews and randomised controlled trials, but the extent of this evidence in physiotherapy has not previously been surveyed. The aim of this survey is to describe the quantity and quality of randomised controlled trials and the quantity of systematic reviews relevant to physiotherapy. The Physiotherapy Evidence Database (PEDro) was searched. The quality of trials was assessed with the PEDro scale. The search identified a total of 2,376 randomised controlled trials and 332 systematic reviews. The first trial was published in 1955 and the first review was published in 1982. Since that time, the number of trials and reviews has grown exponentially. The mean PEDro quality score has increased from 2.8 in trials published between 1955 and 1959 to 5.0 for trials published between 1995 and 1999. There is a substantial body of evidence about the effects of physiotherapy. However, there remains scope for improvements in the quality of the conduct and reporting of clinical trials. PMID- 11869165 TI - Measuring forefoot alignment with a table-mounted goniometric device. AB - Evaluating the need for orthotic treatment may include the measure of forefoot-to hindfoot alignment. This paper describes a table-mounted goniometric device that improves intra-rater reliability and simplifies the measurement of forefoot alignment. Instructions for constructing the device are provided. Use of this device may help clinicians evaluate forefoot alignment when making orthotic correction of the foot. PMID- 11869166 TI - Recent insights into the pathogenesis and therapeutics of pulmonary hypertension. AB - The normal adult pulmonary circulation is a low-pressure, high-capacity circuit. Pulmonary vascular resistance is regulated by alveolar oxygen tension, potassium channels and a variety of locally produced and circulating vasoactive factors. Perturbations of these systems may contribute to the pathogenesis of pulmonary hypertension. Recently, mutations in BMPR2 and ALK-1, genes that encode members of the transforming growth factor-beta (TGF-beta) receptor superfamily, have been found in patients with primary pulmonary hypertension. These observations provide a novel insight into the pathogenesis of primary pulmonary hypertension, and emphasize the importance of the integrity of the TGF-beta receptor family in the maintenance of normal pulmonary vascular structure and function. This review discusses the latest developments in the field of pulmonary vascular biology and the prospects for improving the treatment of pulmonary hypertension. PMID- 11869167 TI - Altered balance of main vasopressor and vasodepressor systems in rats with genetic hypertension and hypertriglyceridaemia. AB - The precise role of nitric oxide (NO) in hypertension is still not fully understood, although this vasodilator system represents the main counterbalance of major pressor systems. The aim of our study was to determine the contributions of superoxide anions, the renin-angiotensin system (RAS), the sympathetic nervous system (SNS) and NO to the maintenance of blood pressure (BP) in Prague hereditary hypertriglyceridaemic (HTG) rats with genetic hypertension. Conscious chronically cannulated rats were subjected to the consecutive blockade of the RAS (losartan, 10 mg/kg), the SNS (pentolinium, 5 mg/kg) and NO synthase [N(omega) nitro-L-arginine (L-NAME), 30 mg/kg]. Some additional rats were pretreated with tempol (a membrane-permeable mimetic of superoxide dismutase). A subsequent genetic study in HTG x Lewis F(2) hybrid rats (n=284) was designed to reveal potential associations of particular BP components with baseline BP. The progenitor study indicated that BP elevation was more pronounced in male than female HTG rats (as compared with normotensive Lewis controls). Higher BP in HTG rats was due to the increased residual BP (measured after combined RAS and SNS blockade) and the augmentation of BP responses to tempol or losartan. In contrast, BP responses to pentolinium or l-NAME were similar in all experimental groups. It should, however, be noted that the baseline BP of progenitor animals was correlated positively with both residual BP and the magnitude of the BP response to pentolinium, but not with BP response to L-NAME. Similarly, the baseline BP of F(2) hybrid rats was positively associated with residual BP, the BP response to pentolinium and the relative SNS contribution to BP maintenance [expressed as a percentage of baseline mean arterial pressure (MAP) values], as well as with the ratio of BP changes elicited by ganglion blockade and NO synthase inhibition (Delta MAP(pentolinium)/Delta MAP(L-NAME) ratio), reflecting the balance of main vasopressor and vasodepressor systems. Thus our studies, performed in progenitor and F(2) hybrid rats, revealed that changes in BP induced by L-NAME do not keep pace with the progressive augmentation of pentolinium induced changes in BP occurring over a wide range of increasing BP. The altered balance between enhanced SNS-dependent vasoconstriction and unchanged NO dependent vasodilation ('relative NO deficiency' in rats with high BP) might result in BP elevation in this form of genetic hypertension. PMID- 11869168 TI - Increases in circulating levels of monocyte-platelet and neutrophil-platelet complexes following hip arthroplasty. AB - Platelets and leucocytes are important effector cells of the haemostatic and inflammatory responses to tissue injury. To investigate the effects of surgical trauma on platelet activation (assessed by measuring levels of P-selectin and beta-thromboglobulin), leucocyte activation (CD11b expression) and leucocyte platelet interactions (leucocyte-platelet complexes), 30 patients undergoing primary hip arthroplasty were studied before and at the end of surgery, and on days 1 and 10 post-operatively, using a whole-blood flow cytometry assay. The inflammatory response was followed by measurement of the levels of C-reactive protein and interleukin-6 in plasma, and the activation of coagulation was monitored by determination of prothrombin fragment 1+2 levels. On day 1 post operatively a significantly increased expression of CD11b on monocytes was noted, but no direct correlation was found between monocyte activation and interleukin-6 production or C-reactive protein at this time point. The percentage of monocyte platelet and neutrophil-platelet complexes was markedly increased on day 10 post operatively compared with pre-operative levels, and levels of these complexes were significantly positively correlated with beta-thromboglobulin levels. Activation of coagulation (prothrombin fragment 1+2) on day 10 post-operatively was positively correlated with the extent of surgical trauma (duration of surgery, amount of blood loss) and with the increase in platelet activation (beta thromboglobulin). In conclusion, hip arthroplasty induces platelet and coagulation activation, and also an inflammatory response that is maintained for more than 10 days post-operatively. This indicates an interaction between the immune and the haemostatic systems in the post-operative phase after hip arthroplasty. PMID- 11869170 TI - The role of inducible nitric oxide synthase in lipopolysaccharide-mediated hyporeactivity to vasoconstrictors differs among isolated rat arteries. AB - We investigated whether organ-specific differences exist in the role of inducible nitric oxide synthase (iNOS) in hyporeactivity to vasoconstrictors following 20 h in vitro exposure of isolated superior mesenteric, renal, hepatic and coronary arteries from the rat to bacterial lipopolysaccharide (LPS). LPS attenuated contraction in response to depolarizing KCl in all arteries. Maximum contractile responses to noradrenaline were attenuated in superior mesenteric and hepatic arteries, and those to the thromboxane A(2) analogue U46619 were attenuated in coronary arteries. LPS shifted the concentration-response curve to noradrenaline in renal arteries to the right. Removal of extracellular L-arginine improved the response to noradrenaline in superior mesenteric and renal arteries only. Addition of the iNOS inhibitor aminoguanidine resulted in full recovery of the responses to noradrenaline in superior mesenteric, renal and hepatic arteries. Contractile responses in coronary arteries did not improve after inhibition of iNOS activity. Therefore the pattern of the LPS-induced changes in vascular reactivity, as well as the contribution of iNOS to impaired vascular constriction, differed among vascular beds. These differences are likely to represent a contributory factor in the sepsis-associated redistribution of cardiac output. PMID- 11869169 TI - Effects of ornithine 2-oxoglutarate on neutrophils in stressed rats: evidence for the involvement of nitric oxide and polyamines. AB - Diets enriched in ornithine 2-oxoglutarate (ornithine alpha-ketoglutarate; OKG) improve immune status during stress. We described previously the ability of OKG to increase the respiratory burst in polymorphonuclear neutrophils (PMNs), but the underlying mechanisms remain unclear. OKG is usually recognized as generating glutamine, arginine and polyamines. The aim of the present study was first to determine the effects of OKG on PMN bactericidal functions (chemotaxis and respiratory burst) in stressed rats, and whether these effects could be reproduced by glutamine- or arginine-enriched diets. Secondly, we investigated the metabolic pathway involved in these actions, using three metabolic inhibitors: methionine sulphoximine (an inhibitor of glutamine synthetase), S methylthiourea (an inhibitor of inducible nitric oxide synthase) and difluoromethylornithine (an inhibitor of ornithine decarboxylase). OKG, arginine and glutamine all increased the production of reactive oxygen species (evaluated by chemiluminescence, ferricytochrome c reduction and flow cytometry). Only OKG markedly enhanced the chemotaxis index (5-fold). Inhibition of glutamine synthetase showed that glutamine production was not involved in the action of OKG. The use of S-methylthiourea and difluoromethylornithine demonstrated that OKG modulated the respiratory burst via nitric oxide (NO*) and polyamine generation. Moreover, OKG stimulated PMN migration via NO*, but arginine administration failed to reproduce this effect. These data suggest that OKG (or its metabolites) and arginine are channelled differently in PMNs. This hypothesis deserves further study. PMID- 11869171 TI - Vascular K(ATP) channel blockade by glibenclamide, but not by acarbose, in patients with Type II diabetes. AB - Glibenclamide inhibits the opening of vascular ATP-sensitive potassium (K(ATP)) channels, which represents a protective mechanism during ischaemia. This effect may imply harmful cardiovascular effects of glibenclamide when used under conditions of ischaemia in patients with Type II diabetes. Acarbose is not associated with effects on the cardiovascular system, because the drug is not absorbed from the bowel. Therefore we hypothesized that treatment of Type II diabetes patients with glibenclamide will impair the vasodilator function of K(ATP) opening, unlike treatment with acarbose. A double-blind randomized cross over study in 12 patients with Type II diabetes was performed to compare the effects of glibenclamide with those of acarbose on the vasodilator responses to K(ATP) channel opening in the forearm vascular bed. The study consisted of two periods: 8 weeks of treatment with orally administered glibenclamide (10 mg x day(-1)) followed by 8 weeks of treatment with acarbose (300 mg x day(-1)), or vice versa. At the end of each treatment period, forearm blood flow (venous occlusion plethysmography) in response to intra-arterially administered diazoxide, acetylcholine and dipyridamole and to forearm ischaemia was measured. The diazoxide-mediated increase in the forearm blood flow ratio (infused/control arm) was significantly less pronounced after glibenclamide than after acarbose (290 +/- 58% and 561 +/- 101% respectively; P<0.0005). Forearm blood flow responses to acetylcholine, dipyridamole and forearm ischaemia were similar during glibenclamide and acarbose treatment. Thus, in patients with Type II diabetes mellitus, treatment with glibenclamide is associated with an attenuated response to K(ATP) opening as compared with treatment with acarbose. This implies that glibenclamide may affect defensive mechanisms under conditions of K(ATP) channel activation. PMID- 11869172 TI - Response of tumour necrosis factor-alpha to delayed in vitro monocyte stimulation in patients with septic shock is related to outcome. AB - We hypothesized that cytokine production following delayed in vitro cell stimulation (to reproduce physiological cellular status at baseline) may be related to outcome in patients with septic shock. A total of 20 patients were included in a prospective clinical study, conducted in a medico-surgical intensive care unit in a university hospital. Blood samples were obtained at the onset of septic shock; these were treated to retain the cells, but to wash out autologous plasma (containing potential inflammatory stimuli such as cytokines, bacterial products and drugs) and replace it with foetal calf serum. Each treated sample was divided into two sets of four aliquots, to be stimulated either immediately or after an overnight period of resting incubation at 37 degrees C. The rest period was to allow recovery from potentially reversible endogenous or pharmacologically induced alterations in cellular response, in order to reproduce a near physiological state at baseline. In vitro cellular challenges used low dose (0.2 ng/ml) or high-dose (1 ng/ml) CD14-dependent lipopolysaccharide and CD14-independent pokeweed mitogen to induce the production of tumour necrosis factor-alpha (TNF-alpha), and interleukins-1 beta and -10. Levels of TNF-alpha, interleukin-1 beta and interleukin-10 were significantly higher (P<0.05) when cell stimulation was delayed for 16 h, indicating a functional down-regulation of cells during septic shock. Moreover, TNF-alpha responses obtained with high-dose lipopolysaccharide were significantly greater in cells from patients who subsequently survived septic shock (n=13; median value 1392 pg/ml; range 592-2048 pg/ml) than in cells from non-survivors (n=7; median value 708 pg/ml; range 520 1344 pg/ml). These observations support the existence of individual differences in the inflammatory response that could influence patient outcome following septic shock. PMID- 11869173 TI - Endothelin-1 inhibits endothelium-dependent vasodilatation in the human forearm: reversal by ETA receptor blockade in patients with atherosclerosis. AB - Several cardiovascular disorders, including atherosclerosis, are associated with endothelial dysfunction and enhanced expression of endothelin-1 (ET-1). The role of ET-1 in the development of endothelial dysfunction in vivo remains unclear. The objective of the present study was to investigate the effect of elevated circulating levels of ET-1 on endothelium-dependent vasodilatation (EDV), and to test the hypothesis that ET(A) receptor antagonism improves EDV in patients with atherosclerosis. EDV and endothelium-independent vasodilatation were determined by brachial artery infusion of acetylcholine and sodium nitroprusside respectively during measurement of forearm blood flow (FBF) with venous occlusion plethysmography. A 60 min intra-arterial infusion of ET-1 (n=10) significantly blunted EDV in young healthy males (33 +/- 13% compared with 271 +/- 74% increase in FBF induced by 10 mug/min acetylcholine; P<0.01). Noradrenaline, which evoked a similar degree of vasoconstriction, did not attenuate EDV. In a separate set of experiments, a 60 min intra-arterial infusion of the selective ET(A) receptor antagonist BQ123 evoked a significant increase in EDV in patients with atherosclerosis (n=10; 109 +/- 45% compared with 255 +/- 101% increase in FBF induced by 10 microg/min acetylcholine; P<0.01), whereas no significant change was observed in healthy age-matched controls (n=9). Endothelium-independent vasodilatation was not affected by ET-1 or BQ123. These observations demonstrate that elevated levels of ET-1 impair EDV in healthy control subjects. Furthermore, ET(A) receptor blockade improves EDV in patients with atherosclerosis, indicating that ET-1 attenuates EDV via an ET(A)-receptor-mediated mechanism. PMID- 11869174 TI - Relationship between cardiomyocyte cell death and cardiac function during hypertensive cardiac remodelling in Dahl rats. AB - The exact mechanisms responsible for the progression of heart failure remain unclear. We investigated the in vivo relationship between the incidence of apoptotic cell death and left ventricular function serially from the beginning of hypertension to decompensated heart failure in Dahl salt-sensitive rats. Dahl salt-resistant and Dahl salt-sensitive rats were fed on a high-salt diet from 6 weeks of age. Systolic blood pressure was recorded by the tail-cuff method every week. Cardiac function in vivo was evaluated by echocardiography and cardiac catheterization. Cardiomyocyte apoptosis was detected by the TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling) method. The gene expression of Bax, Bcl-2 and Bcl-xL was analysed by Northern blotting. The TUNEL method revealed that the incidence of cardiomyocyte apoptosis was significantly increased in the hearts of 18-week-old Dahl salt-sensitive rats (apoptotic index 1.3 +/- 0.1%). Northern blot analysis revealed that the Bcl-xL mRNA level increased gradually during the progression towards heart failure. In conclusion, these data suggest that cardiomyocyte apoptosis is a terminal event, and plays a role as an aggravating factor in the vicious cycle of heart failure. PMID- 11869175 TI - Hepatocyte mitochondrial metabolism is inhibited in neonatal rat endotoxaemia: effects of glutamine. AB - Glutamine has beneficial effects on enterocytes and the immune system in sepsis, but its effects on hepatic metabolism remain unknown. The aim of the present study was to determine the effects of glutamine on hepatocyte energy metabolism under conditions of neonatal endotoxaemia. Suckling Wistar rats were injected intraperitoneally with 200 microg/kg lipopolysaccharide. Oxygen consumption was measured polarographically in hepatocytes respiring on either palmitate (0.5 mM) or palmitate plus glutamine (10 mM). Total hepatocyte oxygen consumption was similar in hepatocytes from control and endotoxic rats, but this was due to a decrease in intramitochondrial and an increase in extramitochondrial oxygen consumption in the cells from endotoxic animals. The addition of glutamine to hepatocytes from endotoxic rats restored intramitochondrial oxygen consumption to control levels. Although glutamine did not reverse the inhibition of the thermogenic proton leak observed in endotoxaemia, it significantly increased oxygen consumption due to mitochondrial ATP synthesis (P=0.03). Glutamine significantly increased the hepatocyte ATP/ADP ratio (P=0.02 compared with hepatocytes from endotoxic rats). Electron microscopy revealed morphological damage to the mitochondria of hepatocytes from endotoxic rats, and a return to a normal appearance with the addition of glutamine. We conclude that glutamine reverses the inhibition of mitochondrial metabolism that is observed in endotoxaemia. The effect is primarily at the level of ATP synthesis. PMID- 11869176 TI - Alcohol consumption, the metabolic syndrome and insulin resistance in 58-year-old clinically healthy men (AIR study). AB - It has been shown that a light-to-moderate intake of alcohol may enhance insulin sensitivity; a decrease in insulin sensitivity is a component of the clustering of risk factors known as the metabolic syndrome. However, previous studies have been limited to relatively small or heterogeneous study groups, or have used suboptimal methods of measuring insulin action. Hence the aim of the present study was to examine whether the metabolic syndrome (as recently defined),components of this syndrome and smoking are associated with alcohol consumption. The study was performed in a population-based sample of clinically healthy men (n=391), all 58 years old and not undergoing any treatment with cardiovascular drugs. Insulin-mediated glucose uptake (euglycaemic hyperinsulinaemic clamp) was measured in a subgroup of these subjects (n=104). Trend analysis showed no difference in alcohol intake across the groups of men with none of the criteria in the definition of the metabolic syndrome (n=77),men with one or more of the criteria (n=252) and men fulfilling all criteria (n=62). However, in the whole group (n=391), alcohol consumption was significantly positively associated with serum triacylglycerols (triglycerides), high-density lipoprotein (HDL) cholesterol and cigarette-years. Furthermore, alcohol consumption was positively associated with insulin-mediated glucose uptake (r=0.20, P<0.05). In multiple regression analyses, body mass index, alcohol consumption and serum triacylglycerols were independent co-variates to insulin mediated glucose uptake. Thus, in 58-year-old healthy men recruited from the general population, there was a significant association between alcohol consumption, serum triacylglycerols, HDL cholesterol and cigarette-years. In a subgroup of 104 subjects, alcohol consumption was independently and positively associated with insulin-mediated glucose uptake. To our knowledge, this is the first study to show an independent relationship between insulin sensitivity, as measured by the clamp technique, and alcohol intake. PMID- 11869178 TI - QT-interval variability and autonomic control in hypertensive subjects with left ventricular hypertrophy. AB - Left ventricular hypertrophy is a risk factor for sudden death. Malignant ventricular arrhythmias originate from altered cardiac repolarization. Ample data have described spatial abnormalities in cardiac repolarization [QT interval (QT) dispersion] in subjects with hypertension; more data are needed on temporal changes. This study was designed to assess the QT variability index (QTVI), the slope between QT and the RR interval (QT-RR(slope)) and spectral QT variability in subjects with arterial hypertension. The results were compared with those from a population at high risk of sudden death, i.e. patients with hypertrophic cardiomyopathy (HCM) who had received an implantable cardioverter/defibrillator (ICD), and those from normotensive control subjects. A total of 44 hypertensive subjects, six patients with HCM and an ICD and 33 control subjects underwent simultaneous short-term recording (256 beats) of QT, RR and systolic blood pressure variability, in the supine position, during controlled breathing. QTVI and spectral components of QT variability in the hypertensive group were significantly higher than in normotensive control subjects (P<0.001), but significantly lower than in patients with HCM and an ICD (P<0.001). The severity of left ventricular hypertrophy correlated significantly with QTVI and the ratio of low-frequency (LF) to high-frequency (HF) power obtained from the RR variability spectra (RR(LF/HF), slope=0.24, P<0.05; QTVI, slope=4.06, P<0.0001; intercept, slope=2.40, P<0.05; chi(2)=38.8; P<0.0001). The QT--RR slope was significantly higher only in patients with HCM and an ICD (P<0.001). In conclusion, the increased QTVI and the correlation of this index with left ventricular hypertrophy indicates that hypertension increases temporal cardiac repolarization abnormalities. At the level of the cardiac sinus node, this alteration is associated with increased sympathetic and reduced vagal modulation. As already noted in patients with HCM, the increased QTVI could be a factor responsible for triggering malignant ventricular arrhythmias in subjects with hypertension. PMID- 11869177 TI - Skeletal muscle mRNA levels for cathepsin B, but not components of the ubiquitin proteasome pathway, are increased in patients with lung cancer referred for thoracotomy. AB - Muscle wasting is a common and prominent feature of advanced cancer, including lung cancer. Evidence from animal experiments suggests that accelerated proteolysis via the ubiquitin--proteasome pathway is the primary cause of cancer related cachexia. However, there are few data on the role of this pathway in determining muscle wasting in human cancer. The present study was designed to measure whether skeletal muscle gene expression of components of the ubiquitin proteasome pathway and/or the lysosomal proteolytic pathway was increased in patients with early lung cancer. A total of 36 patients with lung cancer referred for curative resection and 10 control subjects had biopsies of latissimus dorsi muscle taken at operation. mRNA levels of four components of the ubiquitin proteasome pathway, i.e. polyubiquitin, C2 alpha proteasome subunit, 14 kDa ubiquitin-carrier protein and ubiquitin-activating protein, and of two lysosomal proteolytic enzymes, i.e. cathepsin B and cathepsin D, were measured using quantitative Northern blotting. mRNA levels for cathepsin B, but not for components of the ubiquitin--proteasome pathway, were higher in patients with cancer compared with controls (P=0.01). Among lung cancer patients, cathepsin B mRNA levels correlated with fat-free mass index (r = -0.57, P=0.003) and tumour stage (r(s)=0.45, P=0.03), and were higher in smokers (P=0.04). Thus gene expression of the lysosomal protease cathepsin B is increased in the skeletal muscle of patients with early lung cancer, and the strong inverse relationship with fat-free mass suggests that cathepsin B may have a role in inducing muscle wasting in the early stages of lung cancer. PMID- 11869179 TI - Cystatin C of pleural effusion as a novel diagnostic aid in pleural diseases of different aetiologies. AB - There has been considerable recent interest in the potential use of serum cystatin C as a diagnostic tool. Here we examined the hypothesis that the cystatin C level in the pleural effusion can differ from the corresponding serum level. We evacuated pleural effusion fluids from 47 patients by thoracentesis. Cystatin C, beta(2)-microglobulin, inorganic phosphate, creatinine and total protein were quantified in both pleural effusion fluids and corresponding sera. We determined cystatin C levels in pleural effusions and calculated the ratio of cystatin C levels in serum and effusion, to discriminate between effusions caused by severe renal impairment and other types of effusion. Extremely high concentrations of cystatin C in serum/effusion pairs were only measured in patients with renal failure (6.0 +/- 0.8/6.0 +/- 0.8 mg/l, means +/- S.D., n=11). A clearly defined region was found to correspond to pleural effusion caused by renal failure (r=0.954). The quantification of cystatin C in the effusion was justified by the discovery that there were some patients with a high serum cystatin C level but a low effusion concentration, or a low serum cystatin C but a high effusion concentration, indicating causes other than renal failure. In conclusion, the pilot data indicate a relationship between the cystatin C concentration in pleural fluid and the underlying disease. Thus cystatin C levels in pleural effusion and serum may be a valuable criterion for the differential diagnosis of pleural diseases of different aetiologies. PMID- 11869181 TI - Preservation of fertility in nature and ART. AB - Individuals may regard reproduction as optional but sufficient number of them must be productive to perpetuate the species. The reproductive system is surprisingly vulnerable and depends, among other things, on a limited endowment of oocytes, controlled proliferation of spermatogonial stem cells and the genetic integrity of both. The developmental competence of oocytes and spermatogonial stem cells is maintained by evolved mechanisms for cellular detoxification and genomic stability, and excess or damaged cells are eliminated by apoptosis. Gonadal failure as a result of germ cell depletion can occur at any age, and from the effects of chemical cytotoxicity, disease and infection as well as genetic predisposition. Among extrinsic factors, alkylating agents and ionizing radiation are important causes of iatrogenic gonadal failure in young women and men. In animal models, there is evidence that hormonal manipulation, deletion of genes involved in apoptotic pathways and dietary manipulation can protect against natural and induced germ cell loss, but evidence in humans is absent or unclear. Assisted reproductive technologies (ARTs) provide an ensemble of strategies for preserving fertility in patients and commercially valuable or endangered species. Semen cryopreservation was the first technology for preserving male fertility, but this cannot serve prepubertal boys, for whom banking of testicular biopsies may provide a future option. In sterilized rodents, cryopreserved spermatogonial stem cells can recolonize seminiferous tubules and reinitiate spermatogenesis, and subcutaneous implantation of intact tubules can generate spermatozoa for fertilization in vitro by intracytoplasmic sperm injection. Transplantation of frozen-banked ovarian tissue is well-established for restoring cyclicity and fertility and is currently undergoing clinical evaluation for cancer patients. When restoration of natural fertility is unnecessary or reimplantation is unsafe, it is desirable to culture the germ cells from thawed tissue in vitro until they reach the stage at which they can be fertilized. Low temperature banking of immature germ cells is potentially very versatile, but storage of embryos and, to a lesser extent, mature oocytes is already practised in a number of species, including humans, and is likely to remain a mainstay for fertility preservation. PMID- 11869182 TI - Structural organization and evolution of the marsupial zona pellucida. AB - In this review, the biochemical composition and structural organization of the marsupial and eutherian zonae pellucidae are compared. Differences between the zonae from these two groups of mammals are observed in their response to dilute proteases and reducing agents, in their potential glycosylation patterns, and in some of their functions. However, studies on the glycoconjugates and polypeptides of the three zona pellucida genes have not explained these different responses to the proteases and reducing agents. There is high sequence similarity between the zona polypeptides of marsupials and eutherians, as well as a similarity in the oligosaccharides present, as demonstrated by lectin staining. As the marsupial and eutherian lineages diverged from a common ancestor over 100 million years ago, these observations indicate that the three-dimensional structure of these glycoproteins is highly conserved throughout all mammals, although the complexity of its molecular organization has yet to be resolved. Phylogenetic analyses indicate that there are at least four groups of paralogous zona pellucida genes in vertebrates. The marsupial ZPA and ZPB genes have been named in accordance with their orthologues but the phylogenetic relationships of the marsupial ZPC gene require further investigation. PMID- 11869183 TI - Survival factors regulating ovarian apoptosis -- dependence on follicle differentiation. AB - Only a minute fraction of the ovarian follicles present in a fetal ovary will complete the path to ovulation. Most of the follicles will undergo atresia, a hormonally controlled apoptotic process. Apoptosis occurs at each stage of follicular development and there is a marked reduction in the number of follicles present at birth. Stage-dependent mechanisms of follicle survival can be postulated to achieve co-ordinated development, leading to ovulation of a small fraction of follicles. Indeed, hormone and growth factor regulation of follicular atresia is stage-specific. This short review considers the factors that promote survival of ovarian follicles throughout development, including endocrine, locally produced and intracellular mediators, as exemplified mainly by follicular development in rodents. In primordial follicles, oocyte apoptosis is considered to be the cause of subsequent follicle degeneration. In slow-growing preantral follicles, FSH is not a survival factor, but some locally produced growth factors are. Progression to the antral follicle stage is probably the most critical stage of follicle development in vivo, and FSH is a major survival factor at this stage. In addition, insulin-like growth factor I and interleukin 1beta are potent survival factors for cultured rat follicles at the antral stage. Preovulatory follicles express receptors for LH, and both of the gonadotrophins are survival factors at this stage. Relatively little is known about the period between the LH surge and ovulation; however, it has been suggested that at this stage progesterone acts as a survival factor. PMID- 11869184 TI - Phospholipase C isoforms in mammalian spermatozoa: potential components of the sperm factor that causes Ca2+ release in eggs. AB - Injection of a soluble protein factor from mammalian spermatozoa triggers Ca2+ oscillations in mammalian eggs similar to those seen at fertilization. This sperm factor also generates inositol 1,4,5-trisphosphate and causes Ca2+ release in sea urchin egg homogenates and frog eggs. Recent studies have indicated that the sperm factor may be an inositol-specific phospholipase C (PLC) activity. This study investigated whether any of the commonly known PLC isoforms are components of the sperm factor. PLCbeta, PLCgamma and PLCdelta isoforms were shown to be present in boar sperm extracts. However, upon column fractionation of sperm extracts, none of the PLC isoforms detected correlated with the ability to cause Ca2+ release in eggs. In addition to our previous work on recombinant PLCs, it was also shown that PLCdelta3, PLCdelta4 and its splice variant PLCdelta4 Alt1 fail to cause Ca2+ release. The recently discovered 255 kDa PLCepsilon isoform also appears unlikely to be a component of the sperm factor, as fractionation of sperm extracts on a gel filtration column demonstrated that the peak of Ca2+ releasing activity was associated with fractions of 30-70 kDa. These findings indicate that the sperm factor that triggers Ca2+ release in eggs does not appear to have a known PLC isoform as one of its components. PMID- 11869185 TI - Gap-junctional communication in mouse cumulus-oocyte complexes: implications for the mechanism of meiotic maturation. AB - The mechanisms underlying the hormonal stimulation of meiotic maturation are not understood. The most prevalent hypothesis is that hormone-induced maturation is stimulated by an increase in the intracellular messengers, cAMP or Ca2+. This study investigated whether Ca2+ transients in somatic cells can lead to Ca2+ transients in the oocyte, and whether hormones that stimulate meiotic maturation of mouse oocytes in vitro and in vivo stimulate an increase in intracellular Ca2+. Of a range of potential agonists of Ca2+ release, ATP and UTP were the only agents that stimulated Ca2+ release in cumulus cells. ATP-induced Ca2+ release is from intracellular stores, as the response is not blocked by chelation of extracellular Ca2+, but is inhibited by the Ca2+-ATPase inhibitor, thapsigargin. ATP and UTP are equipotent, consistent with the receptor being of the P2Y2 type. Confocal microscopy was used to show that ATP-induced Ca2+ release in cumulus cells leads to a Ca2+ increase in the oocyte. Inhibition of gap-junctional communication using carbenoxolone, as assayed by dye transfer, inhibited the diffusion of the Ca2+ signal from the cumulus cells to the oocyte. Thus, provided that a Ca2+ signal is generated in the somatic cells in response to maturation inducing hormones, it is feasible that a Ca2+ transient is generated in the oocyte. However, FSH and EGF, both of which stimulate maturation in vitro, have no effect on Ca2+ in cumulus--oocyte complexes. Furthermore, LH, which leads to meiotic maturation in vivo, did not stimulate Ca2+ release in acutely isolated granulosa cells from preovulatory mouse follicles. These studies indicate that ATP may play a role in modulating ovarian function and that diffusion of Ca2+ signals through gap junctions may provide a means of communication between the somatic and germ cells of the ovarian follicle. However, our data are not consistent with a role for Ca2+-mediated communication in hormone-mediated induction of meiosis in mice. PMID- 11869186 TI - Genetic analysis of inferior nurturing ability in RR mice. AB - Females of the inbred mouse RR strain have a limited ability to nurture their offspring, and frequently the young die during rearing. Quantitative trait locus analysis was carried out on the F2 progeny produced from a genetic cross between RR and KK, a strain of normal nurturing ability, to elucidate the putative genetic basis governing certain aspects of the inferior nurturing ability in the RR strain. One hundred and ninety-two F2 female mice were mated with C57BL/6J males. After the number of newborns was adjusted to six per dam, nurturing ability was evaluated on the basis of litter weight at days 7, 12 and 21 after parturition. The results showed that 147 of the 192 F2 dams were able to rear all six pups, although the litter weight varied considerably among litters. Significant evidence for linkage was identified on chromosome 5 near the microsatellite marker of D5Mit161 for litter weight at 12 days after parturition. The R allele (RR strain) at this locus was recessive to the K allele (KK strain), and reduced the litter weight. It is suggested that this locus is one of the heritable components that determine the inferior nurturing ability of RR mice. PMID- 11869187 TI - Effects of ovarian follicle ablation on FSH, oestradiol and inhibin A concentrations and growth of other follicles in sheep. AB - The aim of this study was to examine the effect of removal of the largest follicle or all visible follicles during the first follicle wave on subsequent follicular growth, steroid, inhibin A and gonadotrophin secretion in sheep. On day 4.5 of a synchronized oestrous cycle, ewes (n = 18) were assigned to one of three groups which underwent either no treatment (control), ablation of the largest follicle (largest follicle aspirated and cauterized via laparotomy) or ablation of all follicles (all visible follicles ablated). Between day 0 and day 10 of the oestrous cycle, blood samples were collected every 8 h and ovaries were examined daily using transrectal ultrasonography. The lifespan of the second largest follicle (number of days > 3 mm in diameter) was longer (6.7 +/- 0.9 days; P < 0.05) and the maximum diameter tended to be greater (4.8 +/- 0.3 mm; P = 0.07) in ewes in which the largest follicle was ablated than in the control ewes (3.8 +/- 0.4 days; 4.2 +/- 0.3 mm). There was no difference in the day of emergence of the second follicular wave between groups (day 6.9 +/- 0.4). However, the peak of the transient increase in FSH concentrations after ablation was earlier (day 5.67 +/- 0.15; P < 0.05) in ewes in which all follicles were ablated than in control ewes (day 6.72 +/- 0.36); the timing in ewes that had only the largest follicle ablated was intermediate (day 6.11 +/- 0.28). Serum inhibin A concentrations were about three-fold lower (P < 0.05) in both follicle ablation groups than in the control group. The numbers of follicles 2-3 mm in diameter during the first 3 days of the second follicular wave were greater in 'ablated ewes' (both groups had 2.6 +/- 0.2 follicles day-1) than in control ewes (1.7 +/- 0.3 follicles day-1). It is concluded that: (i) transient increases in FSH concentrations precede the emergence of follicle waves; (ii) ablation of all follicles on day 4.5 after oestrus advanced the timing of the next peak in FSH concentrations and the numbers of small follicles associated with the development of the second follicular wave; and (iii) ablation of the largest follicle resulted in an increase in the lifespan of the second largest follicle, indicating a regulatory role of large dominant follicles over smaller subordinate follicles. PMID- 11869188 TI - Cell death during natural and induced luteal regression in mares. AB - In mares, little information is available on the type of cell death that occurs during natural and induced luteal regression. Corpora lutea were collected from mares in the early luteal phase, days 3-4 (n = 4); mid-luteal phase, day 10 (n = 5); early regression, day 14 (n = 4); late regression, day 17 (n = 4); and 12 and 36 h (n = 3 per group) after PGF2alpha administration on day 10. Histological and ultrastructural sections were examined and TUNEL was used to detect DNA fragmentation. In early luteal regression, there were more pyknotic luteal cells and extracellular round dense bodies compared with the mid-luteal phase. By late regression, there was a significant decline (P < 0.01) in the number of round dense body clusters and a marked accumulation of lipid. Twelve and 36 h after PGF2alpha administration, changes were similar to those seen in natural regression, but there was also a marked infiltration of neutrophils. Accumulation of lipid was not apparent until 36 h after PGF2alpha administration. Ultrastructural examination revealed rarefaction and distortion of the mitochondrial cristae in most of the luteal cells by the mid-luteal phase. Luteal cells showed shrinkage, accumulation of lipid with foamy appearance, and disruption in both smooth endoplasmic reticulum and mitochondria during natural and induced regression. Some luteal cells showed fragmented or pyknotic chromatin characteristic of apoptosis. Other luteal cells showed crenation of the nuclear membrane and shrinkage of the nucleus, features not characteristic of apoptotic cell death. In late regression, capillaries were obstructed by swollen endothelial cells and round dense bodies. These results show that structural regression may be initiated as early as the mid-luteal phase, and is clearly visible by day 14 in natural regression and 12 h after induced regression. Apoptosis did appear to be involved in luteolysis in the equine corpus luteum, but non-apoptotic changes were also observed in some luteal cells during regression. Accumulation of lipid was a late feature of luteal regression. PMID- 11869189 TI - Segregation of spermatozoa within sperm storage tubules of fowl and turkey hens. AB - In avian species, spermatozoa reside in the oviduct for prolonged periods in specialized structures known as sperm storage tubules, but little is known about the relative distribution of spermatozoa in these tubules after successive inseminations by different males. The staining efficacies of various fluorescent dyes for fowl and turkey spermatozoa were evaluated to investigate one proposed mechanism of sperm competition. Hens were then inseminated at different intervals with stained and unstained spermatozoa to observe the spatial distribution of spermatozoa within the storage tubules. Several novel fluorescent lipophilic tracers that successfully stain mammalian spermatozoa either did not stain fowl or turkey spermatozoa, or greatly impaired sperm motility. In contrast, Hoechst 33342 readily stained sperm nuclei (fowl: 25 nmol l-1; turkey: 77 nmol l-1) within 4 h without inhibiting sperm motility, or affecting fertility or the hatching ability of the eggs. Hens were tandemly inseminated with equal numbers of stained or unstained spermatozoa at 24 h intervals and were killed 24 h after the final insemination to study sperm entry and storage within the tubules. Oviductal mucosa containing sperm storage tubules was removed, and individual tubules were classified as containing stained spermatozoa, unstained spermatozoa, a mixture of stained and unstained spermatozoa, or as not containing spermatozoa. Results from the present study indicate that spermatozoa from two different inseminations generally segregate into different storage tubules in both fowl and turkey hens. Storage tubules containing mixed populations of spermatozoa were found in only 4% of fowl and 12% of turkey storage tubules examined. Thus, the mechanism of last-male precedence does not appear to be due to the stratification of spermatozoa within the tubules. PMID- 11869190 TI - Effects of ovarian steroids and epidermal growth factor (EGF) on expression and bioactivation of specific regulators of transcription and translation in oviductal tissue in pigs. AB - The epidermal growth factor receptor (EGF receptor) system is involved in regulation of proliferation and differentiation in oviductal and endometrial tissues. In this study the influence of ovarian steroids and EGF on the expression and activity of specific markers of transcription (mitogen-activated protein kinase; MAP42k) and translation (a potential repressor of eukaryotic initiation factor 4E; 4E-BP1) in pig oviducts was investigated. Furthermore, determination of the distribution of translationally active (polysomal) and repressed (free) mRNA, and cell cycle analysis were performed. Oviductal tissue collected at two points of the oestrous cycle (days 12 and 20) from gilts and tissues from ovariectomized gilts with or without steroid replacement treatment were analysed. The influence of EGF was detected by culture of oviductal explants. MAP42k activity was stimulated by oestrogen treatment, whereas progesterone treatment appeared to decrease its activity. High oestrogen but not high progesterone concentrations resulted in reduced mobility of 4E-BP1 on polyacrylamide gels, indicating its inactivation. EGF and oestrogen treatment of oviductal explants further reduced the mobility of 4E-BP1 on polyacrylamide gels. High concentrations of oestrogen in the plasma promoted cell cycle activity. Progesterone treatment alone did not stimulate the rate of DNA synthesis. There were no significant differences in the distribution of free oviductal poly (A+) mRNA, but the amount of polysomal mRNA was downregulated by oestrogen and progesterone. Increased oestrogen concentrations are involved in the regulation of MAP42k and 4E-BP1 activation in the oviductal tissue of pigs. The effect of oestrogen and EGF in reducing the mobility of 4E-BP1 on gels in oviductal explants indicates that EGF may mediate the effect of oestradiol in the oviducts. PMID- 11869191 TI - Composition and morphology of the follicular basal lamina during atresia of bovine antral follicles. AB - The fate of the follicular basal lamina during atresia was investigated using bovine follicles, in which different follicle phenotypes have been observed. These phenotypes include: healthy follicles with rounded basal granulosa cells with an aligned basal lamina or follicles with columnar basal granulosa cells with a basal lamina of many loops (loopy), and atretic follicles in which either the antral granulosa cells (antral atresia) or the basal cells (basal atresia) die first. Loopy lamina and basal atresia occur only in small antral follicles < 5 mm in diameter. Follicles were collected from cattle of unknown reproductive history and processed for immunohistochemistry and electron microscopy, and from animals in which follicle growth had been monitored by daily measurements of follicle diameter by ultrasonography. Electron microscopic observations of dominant follicles during the growth phase, plateau and regression showed that the basal lamina was still visible and intact upon atresia. These follicles had a conventional aligned basal lamina, which they retained, except for some degree of folding, as they progressed into antral atresia. In small follicles (2-5 mm in diameter), the basal cell shape (rounded or columnar) and appearance of the basal lamina (aligned or of many loops) did not appear to be related to the type of atresia. On atresia the follicular basal laminae retained immunoreactive laminin alpha1 and beta2, type IV collagen alpha1 and nidogen. Laminin alpha2, which may come from the theca, was present in the follicular basal lamina of only 22% of healthy follicles, but was expressed very commonly in 71% of the atretic follicles. Laminin alpha2 expression was found in both phenotypes of healthy follicles, antral and basal atretic follicles, and follicles with aligned or loopy basal laminae. It is concluded that the basal lamina is not degraded upon atresia, but does undergo a variety of other changes. PMID- 11869192 TI - Regulation of endometrial endothelial cell proliferation by oestrogen and progesterone in the ovariectomized mouse. AB - Although the endometrial epithelial and stromal cell response to oestrogen and progesterone is well characterized, relatively little is known about the endothelial cell response. The aim of this study was to investigate the time course of endometrial endothelial cell proliferation in response to a specific regimen of oestrogen and progesterone, and to compare it with the stromal and epithelial cell response in mouse endometrium. Adult female mice were ovariectomized to induce endometrial regression. After 7 days, hormonal treatments were given according to the following regimen: days 1-3: 100 ng oestradiol; days 4-6: 10 ng oestradiol and 500 microg progesterone; and day 7: 100 ng oestradiol and 500 microg progesterone. On each day of hormonal treatment, mice (n = 5) were injected with bromodeoxyuridine and perfusion fixed 4 h later with buffered formalin. Proliferating endometrial cells were detected by monoclonal antibody against bromodeoxyuridine, and endothelial cells were detected by antibody to CD31. At day 7 after ovariectomy few proliferating cells were found in the endometrium. After 1 day of oestrogen treatment, significant proliferation was detected in the endothelial cells (0.0% versus 16.1 +/- 1.2%, P < 0.001). In contrast to the rapid response of the vasculature, glandular epithelial proliferation increased only after 2 days of oestrogen treatment (7.6 +/-1.3% versus 18.8 +/- 2.4%, P < 0.05). Progesterone with low dose oestrogen treatment tended to reduce epithelial and endothelial cell proliferation compared with the effect of high dose oestradiol alone. A combination of progesterone with high dose oestrogen induced higher rates of endothelial cell proliferation than did any other treatment (20.8 +/- 3.2%). These results demonstrate that oestrogen induces rapid proliferation of endometrial endothelial cells, indicating that vascular growth apparently precedes endometrial tissue remodelling. These data also demonstrate that the proliferative response of endometrial endothelial cells to oestrogen and progesterone is different from that of either epithelial or stromal cells. PMID- 11869193 TI - Comparison of uteroplacental glycosylation in the camel (Camelus dromedarius) and alpaca (Lama pacos). AB - The recent birth of a camel-llama hybrid, after numerous failed attempts, has prompted an investigation into the glycosylation of apposing fetal and maternal tissues of pregnant camels and alpacas. This study was undertaken to determine whether interspecies differences in glycans are factors that may account in part for the difficulty in producing a viable hybrid. Specimens of camel placentae from day 60 to day 375 of gestation and alpaca placentae from day 22 to term (approximately 345 days) were fixed and embedded in resin, and sections were stained with a panel of 19 biotinylated lectins and an avidin--peroxidase revealing system. Several qualitative interspecies differences in tissue glycosylation were found, mainly in the trophoblast, and especially with respect to bi/tri-antennary bisected N-glycan, fucosylated structures, beta-galactosyl residues and sialyl termini. In the maternal uterine epithelium, differences were found mainly in bi/tri-antennary bisected complex N-glycan and beta-galactosyl residues, indicating that there is more conservation of glycosylation in maternal tissues compared with trophoblast. There were also many quantitative differences in the distribution of glycans. It is possible that a failure to effect the normal glycan--glycan complementation that occurs at the cell surface between maternal and fetal tissues during the implantation processes of apposition and adhesion may account in part for the difficulty in establishing a viable pregnancy between these two species. PMID- 11869194 TI - Comparative changes in plasma concentrations of progesterone, oestradiol and LH during the ovulatory cycle in a multiple ovulating male line and a single ovulating traditional line of turkeys. AB - The aim of this study was to compare the profile of circulating concentrations of LH, progesterone and oestradiol in a multiple ovulating male line with that of a single ovulating line of traditional turkeys. Plasma samples from seven traditional and 12 male-line turkeys were obtained every 3 h for 36 h. Male-line and traditional turkeys had single peaks of LH and progesterone that were of similar duration in both lines. The mean height of the plasma peaks of LH and progesterone were similar in the two lines and there was no detectable peak plasma oestrogen concentration. Mean plasma concentrations of LH and oestrogen were higher in single compared with multiple ovulating turkeys, whereas there were no differences in mean plasma progesterone concentrations. The results indicate that the multiple ovulation state in genetically selected high-growth lines of turkey may be the result of a correlated response in the steroidogenic capacity of ovarian tissue associated with low plasma concentrations of oestrogen rather than of a disturbance in the hormone profile of the ovulatory cycle. PMID- 11869196 TI - Influence of donor age on development of gonadal tissue from pouch young of the tammar wallaby, Macropus eugenii, after cryopreservation and xenografting into mice. AB - Ovaries from a marsupial, the tammar wallaby (Macropus eugenii), were grafted into a eutherian recipient at known stages of development to ascertain whether normal development would occur. Xenografted ovaries from pouch young < 20 days old, before the onset of meiosis, retained few germ cells and developed tubule like structures reminiscent of seminiferous cords. Ovaries from 50-day-old pouch young, which contain primordial follicles, developed into antral follicles and corpora lutea within the eutherian host, and produced hormones that stimulated the reproductive tract of the host. The timing of onset of antrum formation and the progress of follicle development were advanced relative to the timing of events in ovaries in situ. Frozen-thawed ovaries from 50-day-old donors developed into preantral follicles, but at a reduced rate and number. This finding shows that gonads of a marsupial species can develop as xenografts in a eutherian, forming large antral follicles. Accelerated follicular development in xenografts provides a potentially valuable model for studying the factors that control follicle development. Assisted reproduction of endangered marsupials may also be feasible using follicles from pouch young grown as xenografts in a eutherian host. PMID- 11869195 TI - Possible mechanism for acceleration of meiotic progression of bovine follicular oocytes by growth factors in vitro. AB - The mechanism for the accelerating effects of epidermal growth factor (EGF) and insulin-like growth factor I (IGF-I) on the meiotic cell cycle of bovine oocytes cultured in vitro was investigated. Cumulus-oocyte complexes (COCs) were obtained from small (< or = 3 mm in diameter), medium (4-6 mm in diameter) or large (7-10 mm in diameter) ovarian follicles and cultured with or without a combination of EGF and IGF-I (growth factors). Growth factors significantly increased the frequency of first polar body extrusion of oocytes derived from small follicles at 16 h of culture (PB16 oocytes; with growth factors: 75%; without growth factors: 55%), but did not increase the frequency in oocytes from medium or large follicles. COCs from small follicles were cultured with individual growth factors and sampled for kinase activity. The frequencies of polar body extrusion in EGF only (67%) and EGF + IGF-I (75%) treatment groups were significantly higher than those in the control (no growth factor) group (49%), but not significantly higher than in the IGF-I only group (63%). The H1 kinase activity at 6-8 h of culture in each group increased significantly from the baseline value at 0 h of culture, and the H1 kinase activities in the EGF only, IGF-I only and EGF + IGF-I treatment groups were significantly higher than those in the control group at 8 h of culture. MAP kinase activity was significantly higher than the baseline value and significantly higher than that in the control group at 6 h of culture in the EGF treatment group only. In conclusion, EGF and IGF-I act on COCs from small follicles to accelerate the meiotic cell cycle of the oocytes. This accelerating effect may be related to increased H1 and MAP kinase activities during the early stages of maturation. PMID- 11869198 TI - Minimum number of spermatozoa required for normal fertility after deep intrauterine insemination in non-sedated sows. AB - A fibreoptic endoscope procedure for non-surgical deep intrauterine insemination in non-sedated sows has been reported. However, the endoscope is an expensive and fragile instrument, and is unsuitable for use under field conditions. The aim of this study was to determine the minimum number of spermatozoa required to maintain optimal fertility using a flexible catheter (1.8 m in length, 4 mm in diameter) for deep intrauterine insemination in 2-6 parity non-sedated sows. Crossbred sows were treated with eCG 24 h after weaning and with hCG 72 h later to induce oestrus. Deep intrauterine insemination was performed 36 h after hCG treatment in 117, 126, 60 and 69 sows with 15.0, 5.0, 2.5 or 1.0 x 10(7) spermatozoa in 10 ml, respectively. Weaned sows (n = 147) not treated with hormones and used for standard artificial insemination (AI) (two inseminations per oestrus with 3 x 10(9) spermatozoa in 100 ml) served as controls. The flexible catheter was passed successfully through the cervix into one uterine horn in 95.4% of the sows in an average of 3.7 +/- 0.09 min. Farrowing rates after deep intrauterine insemination with 15 or 5 x 10(7) spermatozoa did not differ from those of the control group (82.9, 76.2 and 83.0%, respectively), but a significant decrease (P < 0.001) was observed in sows inseminated with 2.5 or 1.0 x 10(7) spermatozoa (46.7 and 39.1%, respectively). In contrast, the number of spermatozoa inseminated did not affect prolificacy. Laparotomy revealed that the tip of the flexible catheter reached approximately the anterior third of the uterine horn. Although deep intrauterine insemination was performed in only one uterine horn, the percentages of embryos collected from the tip of both uterine horns 2 days after deep insemination were not significantly different. The results show that in comparison with standard AI, a 20-60-fold reduction in the number of spermatozoa inseminated and an 8-10-fold reduction in the dose volume can be achieved without decreasing fertility when semen is deposited non surgically into the upper first third of one uterine horn. PMID- 11869197 TI - Expression of luteinizing hormone genes in bovine conceptuses. AB - RT-PCR analysis demonstrated that bovine conceptuses at days 16, 23 and 30 expressed LH-beta-like and glycoprotein hormone alpha-like transcript sequences; adult kidney, liver and brain produced predominantly unspliced products. Sequencing of the LH-beta-like fragment (from conceptuses at day 30) indicated complete homology with the published sequence. In addition, ribonuclease protection assay of RNA samples from bovine conceptuses at day 30 with a bovine LH-beta probe revealed the presence of protected molecules that appeared to be full length. Northern blot analysis of total RNA from conceptuses at day 30 failed to demonstrate the presence of LH-beta or glycoprotein alpha subunit transcripts, whereas both transcripts were readily detected in adult pituitary RNA. Administration of hCG into the uterus of heifers from day 14 to day 16 of the oestrous cycle did not affect circulating progesterone concentrations, whereas the same dose increased progesterone concentrations (P < 0.05) when administered intravenously. These results indicate that the early bovine conceptus transcribes genes encoding LH-alpha and -beta subunits, but at a level unlikely to be of physiological consequence. PMID- 11869199 TI - Anogenital and buttock ulceration in infancy. AB - Rashes in the anogenital and buttock region are some of the commonest dermatological problems occurring in infancy. The most frequent causes seen in clinical practice are ulcerating haemangiomas, bullous impetigo and severe irritant contact dermatitis. Other causes include nutritional deficiencies, bullous diseases, trauma, Langerhans cell histiocytoses and inflammatory disorders such as pyoderma gangrenosum and Crohn's disease. This review presents a brief overview of these causes and outlines the recommended management strategies. PMID- 11869202 TI - Basal cell carcinoma of the lip treated with radiotherapy. AB - Basal cell carcinoma (BCC) of the lip is uncommon relative to other cutaneous sites for BCC, such as the central face or scalp. A female predisposition and predilection for the upper lip have been previously documented. A retrospective analysis of patients treated for BCC of the lip was undertaken within the department of Radiation Oncology, Westmead Hospital, Sydney. Twenty patients were identified; 12 women and 8 men. The majority (15/20) had T1 lesions of the upper lip (17/20). Eleven patients were referred for radiotherapy alone and nine for adjuvant radiotherapy following either incomplete excision or local recurrence. With a mean follow up of 36 months no patient has recurred following either definitive or adjuvant radiotherapy. Despite the majority of BCCs of the lip being amenable to surgery fractionated external beam radiotherapy remains an option especially when functional and/or cosmetic concerns are an issue. We present the findings from this small case series and use our findings to illustrate the role of radiotherapy in treating BCC of the lip. PMID- 11869201 TI - Studies in long-pulsed potassium tritanyl phosphate laser for the treatment of spider naevi and perialar telangiectasia. AB - Utilization of a 1-10 msec vascular laser to treat facial vessels to limit purpura has become available in recent years. Its efficacy and morbidity in the treatment of common vascular lesions require exploration. We have assessed the relative outcomes and morbidity from a single treatment session of two patterns of 'difficult to treat' facial telangiectasia - perialar vessels and spider naevi - with such a system. Two prospective clinical studies with long pulse 532 nm potassium tritanyl phosphate (KTP) laser were performed with subjective and independent objective assessment at 3 days, 1 week and 4 weeks in both studies. Perialar vessels showed good to excellent improvement subjectively at 4 weeks in 53% (8/15), spider naevi in 83% (20/24). Pain of treatment was substantially greater treating perialar telangiectasia than when treating spider naevi (P < 0.01). Morbidity was low in both groups with no long-term complications. Despite the clinical end-point of vessel disappearance at time of treatment and good early improvement in the appearance of perialar vessels, the failure rate was significant at 1 month follow up. It would appear that spider naevi behave more predictably, with vessel disappearance at time of treatment most often forecasting a successful outcome. Although not statistically significant, spider naevi probably respond better than perialar vessels to a single treatment with this laser. PMID- 11869203 TI - Review of neurological manifestations in 196 patients with sebaceous naevi. AB - The incidence of neurological abnormalities in patients with sebaceous naevi may not be as high as has been previously accepted. One hundred and ninety-six patients with epidermal naevi of the sebaceous type were examined for clinical neurological abnormalities. Ninety-three per cent of these patients were neurologically normal on examination. Extensive naevi were present in 21% (3/14) of patients with neurological abnormalities and in 5% (9/182) of those without. A centrofacial location of the naevus was found in 21% (3/14) of patients with abnormalities and in 2% (4/182) of neurologically normal patients. A surprising finding was the low yield of abnormalities from neuroimaging in the patients with clinical neurological abnormalities. Imaging was performed in eight of the 14 and was normal in six patients. Despite the lower than expected rate of associated abnormalities and the low yield of abnormal findings from imaging, it is strongly recommended that all patients with sebaceous naevi have a detailed neurological assessment and that imaging be performed on all those in whom clinical abnormalities are demonstrated, as well as on those patients with large naevi involving the centrofacial area. PMID- 11869204 TI - A granuloma annulare-like eruption associated with the use of amlodipine. AB - A granuloma annulare-like drug reaction is a rarely encountered clinical entity. A 64-year-old Caucasian female developed a granuloma annulare-like reaction 13 days after starting amlodipine and cleared within 3 months after drug cessation. The eruption consisted of multiple erythematous pruritic papules, distributed symmetrically over the lateral aspects of the legs and thighs, as well as on both palms. Histology showed focal collagen degeneration and significant interstitial histiocytic dermal infiltrate suggestive of granuloma annulare. We review previously reported cases of granuloma annulare-like drug reactions, in the context of a recently proposed classification for drug-induced interstitial granulomatous reactions. PMID- 11869205 TI - Epidermolysis bullosa simplex Dowling-Meara due to an arginine to cysteine substitution in exon 1 of keratin 14. AB - Epidermolysis bullosa simplex (EBS) is a blistering disorder affecting the basal layer of the epidermis usually inherited in an autosomal dominant fashion. Most cases are caused by mutations in the genes encoding keratin 5 (K5) and keratin 14 (K14) and are characterized by cytolysis within the basal layer of the epidermis. We report a patient manifesting the Dowling-Meara variant of EBS in whom we characterized a cytosine to thymine transition at codon 125 (R125C) in K14. This missense mutation is located at the amino terminus of the helical rod domain of the keratin 14 molecule, resulting in defective pairing with K5, thereby disrupting keratin tonofibril integrity. PMID- 11869206 TI - Two unusual cases of toxic epidermal necrolysis. AB - Two cases of toxic epidermal necrolysis (TEN) are presented. A 27-year-old woman presented with peripherally located targetoid plaques, papules, blisters and lip erosions which began 9 days after 'recreational' use of 'speed' (dexamphetamine and ephedrine) consistent with erythema multiforme major. Three days later she developed widespread lesions with large areas of blistering affecting 40% of body surface area. The diagnosis was revised to TEN. Intravenous cyclosporin led to rapid prevention of new blister formation. A 71-year-old woman, 3 months after commencing amiodarone, developed extensive erythema, blistering and erosions affecting 50% of body surface area, with a maculopapular rash affecting the limbs and extremities consistent with a diagnosis of TEN. She developed septicaemia following bilateral pneumonia with pleural effusions and died 7 days after admission. PMID- 11869207 TI - Primary cutaneous zygomycosis due to Mucor circinelloides. AB - A 62-year-old woman with myelodysplastic syndrome presented with a 4-week history of a large indurated ulcer with a black eschar on the forearm following trauma. On biopsy a diagnosis of zygomycosis was made as broad, sparsely septate, thin walled hyphae were seen in the deep dermis and subcutaneous fat. The zygomycete fungus Mucor circinelloides was cultured from tissue. Further investigation confirmed that the infection was localized to the skin. The 6 x 4 cm lesion was excised and the defect closed with a neurovascular island flap. No other treatment was undertaken. The patient died 6 months later from her haematological disease without recurrence of the fungal infection. PMID- 11869208 TI - Multiple hereditary glomangiomas: successful treatment with sclerotherapy. AB - Glomangiomas are characterized by cavernous vascular channels surrounded by glomus cells. Multiple glomangiomas, although usually painless, can be a few centimetres in size and appear as blue phlebectatic lesions. Surgical excision of multiple glomangiomas can lead to scarring and recurrences. Laser treatment using vascular lasers and CO2 lasers has been useful in small, superficial lesions. We present the successful sclerotherapy treatment of multiple glomangiomas using sodium tetradecyl sulphate in a 59-year-old man who presented with postoperative recurrence of multiple lesions. PMID- 11869209 TI - Merkel cell carcinoma and multiple cutaneous squamous cell carcinomas in a patient with pityriasis rubra pilaris. AB - A 79-year-old female was diagnosed with Merkel cell carcinoma (MCC) and multiple cutaneous squamous cell carcinomas (SCC) occurring on a background of pityriasis rubra pilaris. At the time of initial diagnosis and treatment for upper limb MCC, axillary nodal metastases were clinically evident. In the ensuing months, she developed multiple rapidly progressing SCC and eventually a left arm soft tissue deposit of metastatic MCC. Treatment involved multiple courses of fractionated radiotherapy. The salient clinical features and supporting evidence for this case are presented. PMID- 11869210 TI - Necrobiotic xanthogranuloma occurring within linear morphoea. AB - A 32-year-old woman with linear morphoea of her left arm was noted to have a yellowish atrophic plaque within the linear morphoea. Review of photographs taken on her first visit 3 years previously revealed that the lesion had been present and had increased in size over the 3 years. Necrobiotic xanthogranuloma (NXG) was diagnosed on biopsy. Paraproteinaemia was not detected. The NXG was surgically excised and it has not recurred in 12 months of follow up. PMID- 11869211 TI - Congenital annular erythema persisting in a 15-year-old girl. AB - We present a 15-year-old girl who has had a persisting extensive annular erythematous eruption since birth. Otherwise, she is healthy and developmentally normal. Multiple investigations throughout the years have been normal or negative. Numerous treatments have been tried, but with little success. Both clinical and histological examination fit no known classification. It is most consistent with annular erythema of infancy but could be the first reported case of a new entity. A review of other causes of annular erythema in infancy is presented. PMID- 11869212 TI - Subacute cutaneous lupus erythematosus presenting in a child. AB - A 5-year-old girl presented in summer with an erythematous, scaly annular eruption in a malar distribution. She had no symptoms or signs of systemic lupus erythematosus. A diagnosis of subacute cutaneous lupus erythematosus was made on the basis of the clinical and histological features, positive anti-Ro antibody and a mildly elevated erythrocyte sedimentation rate. All other investigations, including complement studies, were normal. She has responded well to treatment with 0.5% alclometasone ointment and photoprotection. PMID- 11869213 TI - Frontal fibrosing alopecia associated with cutaneous lichen planus in a premenopausal woman. AB - A 45-year-old premenopausal woman presented with an 18-month history of a band like area of fibrosing alopecia affecting the frontoparietal scalp. She also had marked thinning of the eyebrows. The histopathology was consistent with frontal fibrosing alopecia (FFA). Several months later she developed multiple pruritic papules on the wrists and feet. The clinical presentation and histopathology were consistent with cutaneous lichen planus. Although FFA has been reported to occur with mucosal lichen planus this is the first reported case of FFA associated with cutaneous lichen planus. This provides further evidence that FFA is a variant of lichen planopilaris. PMID- 11869214 TI - Acral pityriasis lichenoides. AB - Two patients presented with acrally located papulosquamous lesions that were polymorphic and had an identical morphology to those of pityriasis lichenoides. The lesions remained localized to the feet and ankles for years. Multiple skin biopsies had the histopathological features of pityriasis lichenoides. In one biopsy syringotropic lymphocytes and early syringolymphoid hyperplasia were observed. This rare variant of pityriasis lichenoides may be under-recognized and misdiagnosed as it resembles a variety of papulosquamous disorders particularly psoriasis. PMID- 11869215 TI - Venison contact dermatitis. AB - A 37-year-old meat inspector presented with a 5-month history of forearm eczema. His symptoms improved at weekends and over holidays but flared within a day of work. Scratch patch tests were strongly positive to fresh venison diaphragm and venison liver and weakly positive to venison hide and blood, and lamb blood. They were negative to other venison and lamb components. Type IV allergy tests with Standard European and fragrance batteries were negative. The eczema cleared with a change in work role, clobetasol-17-propionate cream, flucloxacillin and aqueous cream. He was then able to return to meat inspecting. Provided he was diligent about applying 'Dermashield' barrier foam and minimizing contact with irritants, his eczema has remained well controlled over the last 4 years. Attention to irritant contact dermatitis is important in the management of protein contact urticaria. PMID- 11869216 TI - The results of photopatch testing 172 patients to sunscreening agents at the photobiology clinic, St Vincent's Hospital, Melbourne. PMID- 11869219 TI - Australasian college of dermatologists: conferring of diplomas and awards Adelaide - Sunday 20 May, 2001. PMID- 11869220 TI - Research methods in human respiratory pharmacology. PMID- 11869221 TI - Classification and criteria of otitis media. PMID- 11869222 TI - New views on the pathogenesis of acute otitis media and its complications. AB - The pathogenesis of acute otitis media is complex and multifactorial. Bacteria infecting the middle ear come from the nasopharynx via the eustachian tube. This colonization is facilitated by bacterial adherence on the pharyngeal and the eustachian tube cells. Otitis media is characterized by inflammation of the middle ear with an infiltration of the subperiosteal space by leukocytes, macrophages and mast cells. The effusion contains great amounts of inflammatory mediators (eicosanoids, cytokines, histamine). Elimination of the effusion and/or the bacteria is based on non-specific factors such as mucociliary clearance and phagocytosis, and on a specific immune response which apparently is not the same for Haemophilus influenzae and for Streptococcus pneumoniae. The main complications of acute otitis media are otitis media with effusion, mastoiditis, sensorineural hearing loss and meningitis. PMID- 11869223 TI - Incidence and risk factors of acute otitis media in children. AB - Acute otitis media (AOM) is the most common disease requiring antibiotic treatment in young children. This article reviews the incidence of AOM in different age groups of controlled populations, its incidence in pediatricians, consulting rooms and emergency services and the main circumstances that usually complicate the diagnosis (circumstances of examination, methods and devices used and pediatric skills). Host and environmental factors related to otitis- prone children are revisited. Sex predominance (males), ethnicity, birth order (second born or siblings), early occurrence of infection (when earlier, worse), and sibling history of severe or recurrent ear infections are host factors related to recurrent otitis. Season (inconclusive), socio-economic status (inconclusive) smoke and air pollutants (clearly related), breast-feeding (clearly related, especially when prolonged) and type of day care (the greatest incidence in children in large day-care groups versus home-cared) are environmental factors related to recurrent otitis. PMID- 11869224 TI - Microbial etiologies of acute otitis media. AB - The bacterial etiology of acute otitis media (AOM) in children is well known; Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis account for up to 80% of the cases. Negative cultures are obtained from 15 to 34% of the middle ear effusions obtained from patients with AOM and could represent non-viable bacterial organisms, Chlamydia, Mycoplasma, anaerobes and, perhaps, new and unknown organisms. Viruses are rarely involved as a cause of AOM but upper respiratory tract infections by respiratory syncitial virus, adenovirus, enterovirus, influenza virus and rhinovirus usually precede AOM episodes. Other organisms isolated from middle ear effusions are identified as coagulase-negative staphylococci, alpha---haemolytic streptococci and diphtheroids and could be responsible for AOM; thus these organisms should not always be disregarded and considered as contaminants. PMID- 11869225 TI - Current patterns and evolution of antibiotic resistance among bacterial pathogens involved in acute otitis media. AB - Penicillin resistance among Streptococcus pneumoniae is becoming a global problem. Different evolutionary phases in the development of resistance can be differentiated according to the local distribution of bacterial strains with different levels of penicillin resistance: susceptible, early phase, low-level resistance phase, multipopulational phase, high-level resistance phase and very high-level resistance phase. These phases evolve in different periods (cryptic, penetration period, evolutionary period and stationary period) which describe the presumed natural history of the resistant organisms. The selective effect of the antibiotic usage depends on the amount and type of the drug, and also on the evolutionary period dominating the geographic area where the antibiotic is used. Macrolide resistance is also a growing problem, probably proportional to the amount of drug used. In the case of beta-lactams, 'more is different' strategies may be useful to control clinically low-level resistant strains with aminopenicillins, and could slow the evolution to higher rates of resistance in some areas; nevertheless, surveillance of the evolution of high-level resistance is mandatory. In Haemophilus influenzae, the proportion of beta-lactamase positive strains is also very high, frequently over 25% of cases; in all cases the combination amoxycillin---clavulanate restores the susceptible phenotype. In this case, surveillance should be focused on the increase in non-beta-lactamase mediated penicillin resistance and fluoroquinolone resistance, or in the potential evolution towards extended-spectrum beta-lactamases. PMID- 11869226 TI - Acute childhood otitis media: the diagnostic value of bacterial samples from the nasopharynx. AB - OBJECTIVE: To compare bacteriologic results in acute otitis media (AOM) from middle ear otorrhea and rhinopharyngeal samples. METHODS: Three hundred and fifty four children suffering from AOM were included. Two samples were obtained for each patient, one in the middle ear fluid, and another in the rhinopharynx. RESULTS: The rhinopharyngeal sample shows a poor predictive value for the pathogen responsible for the AOM (only 30%). But antibiotic sensitivity from each bacteriologic sample (otorrhea and rhinopharynx) for the whole population showed a very good correlation. CONCLUSIONS: The collection of nasopharyngeal cultures should therefore be promoted for their collective epidemiologic value, especially with reference to the monitoring of pneumococcal susceptibility in children. PMID- 11869227 TI - Pharmacokinetics of antibiotics in the middle ear. AB - The middle ear mucosa is described as a non-specialized site where drug substances diffuse passively. However, precise pharmacokinetic studies on antibiotic penetration into middle ear effusion (MEE) are difficult to conduct in humans because of ethical and methodological problems: (1) studies must involve the population in which acute otitis media (AOM) is most frequent (i.e. in children from 3 months to 3 years of age); (2) studies are commonly performed during surgery for chronic otitis media with effusion, whereas drug penetration into MEE is higher during AOM; (3) kinetics are deduced from data per individual obtained in groups with marked interindividual variations. Furthermore, the results of different studies are difficult to compare because of differences in protocols, antibiotic assay methods and interpretation. Clinical studies have nonetheless shown the poorly predictive nature of serum concentrations, given the large differences in drug delivery to the MEE among antibiotic classes, among members of the same class, and among individuals receiving a given antibiotic. Experimental studies can help to narrow the gap between in vitro and clinical data, as the more precise pharmacokinetic data they yield can throw light on the link between drug/host/pathogen factors and bacteriologic efficacy. PMID- 11869229 TI - Acute otitis media unresponsive to treatment. AB - Unresponsive acute otitis media is characterized by clinical signs and symptoms associated with otoscopic findings of inflammation that continue beyond 48 h of therapy. This occurs in about 10% of children treated initially with a 10-day course of antibiotics. This article reviews the management of this condition. PMID- 11869228 TI - Antibiotic treatment in acute otitis media: 'in vivo' demonstration of antibacterial activity. AB - The ultimate goal in antibiotic treatment of acute otitis media (AOM) is eradication of the pathogens from the middle ear fluid (MEF). To test the ability of an antibiotic to achieve this goal, the best available method is to obtain MEF by aspiration before initiating antibiotics and to repeat this procedure during treatment, a method which was termed the 'in vivo sensitivity test'. This advantageous method is difficult to perform and thus only a few groups were able to conduct such large-scale studies. The available information is discussed in the present manuscript, as well as some preliminary results suggesting that oral beta-lactam antibiotics may not be the ideal drugs to date, due to the emergence of resistant pneumococci. PMID- 11869230 TI - Practical management of recurrent acute otitis media. AB - Recurrent acute otitis media (RAOM) represents a frequent challenge for the primary care physician and a major source of consultation to otolaryngologists. We recommend insertion of ventilation tubes when school performance or speech acquisition is compromised. Chemoprophylaxis with trimethoprim---sulfamethoxazole (TMP/SMX) is still a viable option in other cases, despite the potential development of drug resistance. Other medical treatments are available, including vaccination and steroids. Finally, we have used limited mastoidectomy in carefully selected cases where retention of infected secretions and lack of aeration are implicated. PMID- 11869231 TI - Prevention of acute otitis media. AB - Breast-feeding, using family or small-group day care for infants and toddlers and avoiding exposure to household tobacco smoke are the main preventive measures against acute otitis media (AOM). It is also useful to immunize children who have recurrent otitis media with the influenza and the pneumococcal vaccines. Antibiotic prophylaxis is the most effective method to reduce the frequency of new episodes of otitis in children with recurrent AOM, but it should be used with caution. Tympanostomy tube placement and/or adenoidectomy can be considered as options in some situations. Now, probably, the best initial steps to take to prevent new episodes of otitis in children with recurrent AOM are antimicrobial treatment of each individual AOM episode and, in certain cases, antibiotic prophylaxis for short periods when an upper respiratory tract infection is present. PMID- 11869232 TI - The role of pneumococcal and haemophilus type B vaccination in the prevention of acute otitis media. AB - Acute otitis media is an extremely common disease, most children being subject to it once or more during their first 5 years of life. Thirty to forty per cent of episodes of acute otitis media are caused by Streptococcus pneumoniae. Resistance of pneumococci to antibiotics is an increasing problem particularly in France. Since 1983 a 23-valent pneumococcal polysaccharide vaccine has been available, containing in particular four serotypes that carry penicillin resistance. Unfortunately, this vaccine is therefore not fully effective until after the age of 2 years. In spite of good serologic responses, clinical results have been variable. Some found that this diminished frequency only lasted for several months and ended as serotypes not included in the 23-valent vaccines began to appear. Since 1992, studies have been carried out using different pneumococcal vaccines conjugated to diphtheria, tetanus or meningococcal proteins. Despite the incomplete nature of the data, it can already be confirmed that these vaccines are immunogenic from the very first months of life onwards. Numerous problems nevertheless remain unsolved. How many serotypes can be included in the conjugate vaccine? For the moment it seems technically impossible to conjugate all 23 serotypes of the current vaccine. While Haemophilus influenzae type b is principally responsible for invasive infections, it is traditionally said to play only a minor role in otitis, which is usually provoked by non-capsulated Haemophilus strains beyond the vaccine's reach. In fact, in most studies it has been observed that 2--5% of cases of Haemophilus otitis are due to Haemophilus type b. The vaccine could be said, therefore, to have a minor but by no means negligible role in the prevention of Haemophilus otitis. In conclusion, pneumococcal and haemophilus vaccines have not revolutionized the preventive treatment of otitis for the moment, but great hopes are still placed on the conjugate pneumococcal vaccine. PMID- 11869233 TI - Decision-making in otitis media in children, Part I: epidemiologic data and definitions for a reliable cost-effectiveness analysis. AB - There is a gap between the medical and economic weight of acute otitis media and the imprecise knowledge and irrational attitude towards diagnosis and treatment. The wide variations among prescribers need to be analyzed. Precise definitions are proposed for acute otitis media and endpoints for outcomes being estimated. An empirical antibiotic treatment gives a greater cure rate than a conservative approach with antipyretic and analgesic drugs. The incremental efficacy expected from more extended-spectrum antibacterials is not clinically significant. PMID- 11869234 TI - Decision-making in otitis media in children, Part II: A cost-effectiveness analysis. AB - Acute otitis media (AOM) is a source of considerable expense of money and care. Two protocols for the empirical treatment of AOM are compared by a cost effectiveness analysis. In protocol 1, treatment is initiated by a standard spectrum antibiotic (amoxicillin) for 5--7 days; in case of clinical failure, a second treatment is prescribed with a wide-spectrum antibiotic (cefaclor) for 7- 10 days. In protocol 2, treatment with a standard-spectrum antibiotic is prescribed only in case of clinical failure after a 'masterful inactivity' for 3- 5 days, then checked on days 5--7, for a prescription of a wide-spectrum antibiotic in case of failure. The two protocols have a similar effectiveness, but the cost of protocol 2 is significantly higher because of a greater expense for medical visits. A low compliance rate is a definite factor of high cost effectiveness ratio. PMID- 11869235 TI - Current trends in the antibiotic therapy of uncomplicated Acute Otitis Media in children: concluding remarks. PMID- 11869236 TI - Preface. PMID- 11869237 TI - beta-Lactamases: quality and resistance. AB - beta-Lactamase-mediated resistance to beta-lactam antibiotics is a feature of great clinical significance. beta-Lactamases are a diverse group of bacterial enzymes that vary in their abilities to hydrolyze beta-lactam antibiotics. beta Lactamases possess an active site containing either a serine moiety or a zinc atom; serine beta-lactamases are currently of greater clinical prevalence. This review considers the molecular classification of beta-lactamases, the structure of the serine beta-lactamase active site and the mechanisms by which beta lactamase production may become derepressed. The spread of beta-lactamases in the clinical setting and some important structural mutations that have extended the hydrolysis profiles of serine beta-lactamases are also discussed. PMID- 11869238 TI - beta-Lactamases: quantity and resistance. AB - Rates of enzyme-mediated catalysis are proportional to enzyme quantity, so increasing the amount of beta-lactamase should increase resistance. Kinetic considerations support this argument for both Gram-positive and Gram-negative bacteria. Direct relationships between resistance and enzyme quantity are most obvious with constitutive beta-lactamases, e.g. the TEM types from Gram-negative bacteria. As the level of TEM enzyme rises, so do the MICs of substrates and the concentrations of inhibitors required to potentiate these substrates. The position is more complex for inducible beta-lactamases, e.g. the AmpC types of Enterobacter spp., Citrobacter freundii, Serratia spp. and Pseudomonas aeruginosa. Third-generation cephalosporins are labile to these enzymes, but are only weak inducers, so the beta-lactamase-inducible strains appear susceptible. Once, however, the enzyme is derepressed, resistance is apparent. The behavior of inhibitor combinations against inducible beta-lactamases is complicated by the propensity of some inhibitors to induce further enzyme synthesis. As the inoculum is raised in laboratory tests, the amount of beta-lactamase and MICs also rise. This is notorious for staphylococci but is seen also for some Gram-negative organisms, notably klebsiellae with extended-spectrum beta-lactamases. A beta lactamase-related inoculum effect generally predicts clinical failure. PMID- 11869239 TI - A comparative study of the in vitro activity of meropenem and representatives of the major classes of broad-spectrum antibiotics. AB - OBJECTIVE: To compare the in vitro activity of meropenem with that of other agents with a broad-spectrum of antibacterial activity, and which may therefore be candidates for empirical use. The agents tested were imipenem, third generation and newer cephalosporins, penicillins combined with a beta-lactamase inhibitor, ciprofloxacin and amino-glycosides. METHODS: Using agar dilution methods, all agents were tested against 900 clinical isolates (many of which were multiresistant), including Gram-positive aerobes, nutritionally fastidious aerobes, Enterobacteriaceae, non-fermenters and anaerobes, collected from 17 UK hospitals during 1994. In addition, some agents were tested against strains expressing defined beta-lactamases, including extended-spectrum beta-lactamases. RESULTS: The potency and spectrum of the carbapenems, unequalled against aerobes and anaerobes, were undoubtedly influenced by their stability to serine beta lactamases. Meropenem and imipenem exhibited essentially the same spectrum of activity but imipenem was often less potent, notably against Gram-negative aerobes, including Pseudomonas aeruginosa and Burkholderia cepacia. Conversely, the activity of the third-generation (MIC90s 0.016--64 mg/L) and, to some extent, the newer cephalosporins (MIC90s 0.06--8 mg/L) and the augmented penicillins (MIC90s 1 to >128 mg/L) was unreliable against many genera of Enterobacteriaceae because of chromosomally mediated enzymes or the, now commonplace, plasmid mediated beta-lactamases. Ciprofloxacin had modest activity (MIC90s 1--64 mg/L) against Gram-positive aerobes, was potent against nutritionally fastidious species, had again variable activity against the Enterobacteriaceae (MIC90s 0.008 -4 mg/L) and was inactive against many strains of Pseudomonas, Burkholderia and Acinetobacter, resulting in MIC90s of 4 to >128 mg/L. The aminoglycosides were impressive only against the Enterobacteriaceae, with amikacin alone active (MIC90s 2--8 mg/L) against the 11 species tested. CONCLUSIONS: This study demonstrates that it is difficult on grounds of spectrum to differentiate third generation cephalosporins, and that neither cefepime nor cefpirome materially enhance utility. The study suggests also that, judged on activity in vitro, meropenem or imipenem are the only monotherapy options for empirical antibacterial therapy of polymicrobic infections or when local epidemiology indicates the predominance of multiresistant Enterobacteriaceae. Instability to current and emerging beta-lactamases is progressively compromising the use of all other beta-lactam compounds. PMID- 11869240 TI - A compilation of studies assessing the in vitro activity of meropenem and comparators in 84 laboratories throughout Europe. AB - OBJECTIVE: To compile the results of investigations conducted in 84 centers throughout Europe of the in vitro activity of meropenem and a standard set of comparators against some 12 000 Gram-positive, Gram-negative, aerobic and anaerobic bacteria. METHODS: Recent clinical isolates from 84 European countries were tested using Mueller---Hinton broth or agar (aerobes and nutritionally fastidious species) or Wilkins---Chalgren medium (anaerobes) for susceptibility to meropenem, imipenem, cefotaxime, ceftazidime, piperacillin, ciprofloxacin and gentamicin (aerobes) or meropenem, imipenem, clindamycin and metronidazole (anaerobes). RESULTS: Whether compared across studies or across countries, meropenem and imipenem were the only compounds that produced uniform and predictable activity. The activity of ceftazidime and cefotaxime was compromized because of instability to both chromosomally mediated and new plasmid-mediated (transferable) beta-lactamases. This effect was even more pronounced with piperacillin. Resistance to gentamicin was commonplace, although variable between countries; strains resistant to ciprofloxacin occurred in many genera of Enterobacteriaceae and amongst the pseudomonads. Metronidazole was uniformly active against anaerobes but there were many clindamycin-resistant strains. Resistance, or diminished susceptibility, amongst Streptococcus pneumoniae was found frequently, but of the beta-lactams tested, only ceftazidime and piperacillin produced minimum inhibitory concentration values that could compromize therapeutic efficacy outside the central nervous system. CONCLUSIONS: These data provide further evidence of the high incidence of antibiotic resistance in Europe, notably in France, Spain and Italy. Fortunately, the carbapenems are highly stable to the enzymes that hydrolyze cephalosporins and penicillins. Hence, they retain an essentially unaltered and exceptional antimicrobial spectrum, embracing the vast majority of strains resistant to the other comparators, although meropenem was more reliable than imipenem against Pseudomonas aeruginosa. PMID- 11869241 TI - Meropenem: laboratory and clinical data. AB - The usefulness of many beta-lactams, including the third-generation cephalosporins, has been threatened in recent years by the dissemination of beta lactamase-mediated resistance. The carbapenems, meropenem and imipenem/cilastatin, demonstrate ultra-broad spectra of antibacterial activity which encompass the majority of clinically significant Gram-positive and Gram negative aerobic and anaerobic pathogens. Due to the unique arrangement of the moieties around the carbapenem beta-lactam ring, these agents possess unrivaled stability to almost all serine beta-lactamases, including the mutant extended spectrum types produced by Enterobacteriaceae which hydrolyze third-generation cephalosporins. Meropenem has a number of additional structural features which confer advantages over imipenem, notably stability to metabolism by dehydropeptidase-I and enhanced activity against Gram-negative organisms, including Pseudomonas aeruginosa. In the treatment of nosocomial respiratory tract infections, meropenem monotherapy has compared favorably with ceftazidime plus aminoglycoside combination regimens or imipenem/cilastatin, producing rates of clinical efficacy of > 75%. Likewise, in patients with intra-abdominal infections, meropenem has proved to be as efficacious as clindamycin plus tobramycin, cefotaxime plus metronidazole and equivalent dosages of imipenem/cilastatin. Meropenem is well tolerated by the central nervous system and, unlike imipenem/cilastatin, is approved for the treatment of meningitis. PMID- 11869242 TI - Therapy of sepsis. AB - The term 'sepsis' is often used synonymously with 'infection' or 'bacteremia'. Additional definitions, e.g. 'systemic inflammatory response syndrome', have been proposed. These terms have been employed to establish entry criteria for clinical trials of antibiotic therapy and adjunctive therapies. It may be appropriate to utilize these descriptions when initiating therapy of suspected bacterial infections, since culture confirmation is often not available for days after initial clinical presentation. Empiric antibiotic therapy is now the rule rather than the exception for febrile hospitalized patients. No antimicrobial agent is superior for all major clinical situations and thus history and physical examination are critical factors which guide the clinician. Guidelines have been derived for the initial empiric management of serious infection. While these were initially refined for the immunocompromized host, they may be considered generally applicable for the hospitalized patient with a suspected serious systemic bacterial infection. Currently, monotherapy with agents such as cefepime, ceftazidime, imipenem/cilastatin and meropenem has been sanctioned on the basis of their broad antimicrobial coverage and stability to clinically important beta-lactamases (although there is considerable variation from one compound to another). Perception of local epidemiologic patterns is a major factor in the selection of initial antibacterial therapy for hospital-acquired infection. PMID- 11869243 TI - Introduction: Evolving needs in respiratory tract infections. AB - Two issues that have become clinically relevant to the treatment of pneumonia over the past few years are the development of antibiotic resistance among respiratory pathogens and the increasing importance of the atypical respiratory pathogens---Mycoplasma pneumoniae, Chlamydia pneumoniae and Legionella spp. Resistance has become an important issue in Streptococcus pneumoniae, methicillin resistant Staphylococcus aureus and Gram-negative rods. The ways by which bacteria become resistant to antibiotics include production of antibiotic modifying enzymes, reduced access to target sites, efflux of antibiotic, change in the bacterial target site and the bypassing of inhibited pathways. In Streptococcus pneumoniae that are penicillin resistant, the mechanism is through alteration of the target site for penicillins (penicillin-binding proteins) and this may also confer resistance to some cephalosporins. Multidrug resistance has also been reported in some strains of pneumococci. Of particular concern is resistance to macrolides mediated by the ermAM gene, which also confers resistance to lincosamides and streptogramin-B drugs. In Staphylococcus aureus, resistance to virtually all beta-lactam drugs is mediated by acquisition of the mecA gene, which codes for the drug-resistant beta-lactam target PBP2a. Antimicrobials are now needed that have enhanced activity against aerobic Gram negative rods, atypical respiratory pathogens and Gram-positive cocci. PMID- 11869244 TI - Grepafloxacin: microbiological properties. AB - Grepafloxacin is a new broad-spectrum fluoroquinolone characterized by having a methyl-substituted piperazine at the 7 position. It is a water-soluble racemate with both stereoisomers having the same activity. Its mode of action involves inhibition of topoisomerases II and IV. Grepafloxacin is not recognized by the NorA efflux mechanism in Staphylococcus aureus and, thus, some strains of Staphylococcus aureus that are resistant to other fluoroquinolones remain susceptible to grepafloxacin. Grepafloxacin has potent in vitro activity against streptococci and staphylococci, respiratory Gram-negative pathogens, atypical respiratory pathogens and sexually transmitted disease pathogens. It combines the positive properties of the beta-lactams against conventional Gram-positive and Gram-negative respiratory pathogens with the activity of the macrolides against the atypical pathogens. Unlike macrolides, it is bactericidal at concentrations close to the MIC. Its in vitro activity has been reflected in animal models of respiratory tract infections. Concentrations above MICs are maintained throughout nearly all of the 24-h dosing interval. Grepafloxacin provides important improvements over older quinolones and over other classes of antibiotics. PMID- 11869245 TI - Structure of grepafloxacin relative to activity and safety profile. AB - A comparison of the structure of ciprofloxacin and grepafloxacin shows that the two compounds are similar, with two exceptions: grepafloxacin has a methyl group at the 5 position and a methyl group attached to the 7-piperazinyl substituent. At the 1 position, both compounds have a cyclopropyl group, which is important for potency, but limits anaerobic activity. The methylpiperazine at position 7 in grepafloxacin is associated with its enhanced Gram-positive activity and long half-life. The methyl group at R5 is also thought to enhance Gram-positive activity. Ciprofloxacin's piperazine group at the 7 position is associated with good Gram-negative activity. Grepafloxacin's Gram-negative activity is comparable to that of ciprofloxacin's against Haemophilus influenzae, Moraxella catarrhalis and enteric Gram-negative bacilli. Studies of resistance development to fluoroquinolones suggest that grepafloxacin is associated with a reduced selection of resistance in Staphylococcus aureus, which is possibly related to the inhibition or avoidance of efflux transport by NorA. PMID- 11869246 TI - Grepafloxacin: pharmacokinetics and tissue penetration. AB - Pharmacokinetic and tissue penetration studies of grepafloxacin, a new broad spectrum fluoroquinolone, show that it has useful properties for the treatment of respiratory tract infections. Grepafloxacin has a volume of distribution that is larger than those of many of the other fluoroquinolones and is concentrated in alveolar macrophages, bronchial mucosa and epithelial lining fluid to a greater extent than are certain other fluoroquinolones. Grepafloxacin concentrations achieved in plasma after a 400-mg oral dose are well in excess of those required to inhibit the respiratory pathogens Staphylococcus aureus, Haemophilus influenzae and Moraxella catarrhalis. Streptococcus pneumoniae is also covered for most of the dosing interval, but at normal dose levels grepafloxacin might not inhibit Enterococcus faecalis. The maximum plasma concentrations and area under the concentration---time curve achieved with grepafloxacin suggest that it will be effective for the treatment of community-acquired pneumonia and acute exacerbations of chronic bronchitis. The pharmacokinetics of fluoroquinolones are among their most useful properties. The aim of this paper is to demonstrate whether the differences between grepafloxacin and the other fluoroquinolones are of therapeutic significance. PMID- 11869247 TI - Clinical efficacy and tolerability of grepafloxacin in lower respiratory tract infection. AB - Studies in community-acquired pneumonia (CAP) have compared grepafloxacin, 600 mg o.d. for 7--10 days, with amoxycillin, 500 mg t.d.s., cefaclor, 500 mg t.d.s., or clarithromycin, 250 mg b.d. Grepafloxacin appeared to be clinically as effective as the comparators. In CAP caused by Haemophilus influenzae, grepafloxacin was significantly superior to amoxycillin (p=0.005) and cefaclor (p=0.003) and equivalent to clarithromycin in eradicating the infecting organism. Bacterial eradication with grepafloxacin in CAP caused by Moraxella catarrhalis or Streptococcus pneumoniae was effective and equivalent to the comparator antibiotic. In an open study, grepafloxacin was also effective in treating atypical pneumonia caused by Mycoplasma pneumoniae and Legionella pneumophila. In acute bacterial exacerbations of chronic bronchitis (ABECB), studies have found once-daily treatment with grepafloxacin, 400 mg or 600 mg for 7--10 days, to be equivalent to amoxycillin, 500 mg t.d.s., or ciprofloxacin, 500 mg b.d. In patients with documented infections, bacteriologic eradication with grepafloxacin, 400 mg or 600 mg o.d., was superior to amoxycillin, 500 mg t.d.s. Results from clinical trials so far indicate that grepafloxacin has a broad spectrum of activity covering all important community-acquired respiratory pathogens, and may be suitable for the empirical treatment of respiratory infection. PMID- 11869248 TI - Summary. PMID- 11869249 TI - Immunopathology of respiratory infections. PMID- 11869250 TI - The etiology of respiratory tract infections and the antibacterial activity of fluoroquinolones and other oral antibacterial agents against respiratory pathogens. PMID- 11869251 TI - Epidemiology and mechanisms of resistance among respiratory tract pathogens. PMID- 11869252 TI - Pharmacokinetics and pharmacodynamics of fluoroquinolones. PMID- 11869253 TI - Lower respiratory tract infections: etiology, current treatment, and experience with fluoroquinolones. PMID- 11869255 TI - Conclusions. PMID- 11869254 TI - Upper respiratory tract infections: etiology, current treatment, and experience with fluoroquinolones. PMID- 11869256 TI - Introduction. PMID- 11869257 TI - Approach to diagnosis of infective endocarditis. PMID- 11869258 TI - Microbiological recommendations for the diagnosis and follow-up of infective endocarditis. PMID- 11869259 TI - Antibiotic treatment of infective endocarditis due to viridans streptococci, enterococci, and other streptococci. PMID- 11869260 TI - Staphylococcal endocarditis---recommendations for therapy. PMID- 11869261 TI - Recommendations for the surgical treatment of endocarditis. PMID- 11869262 TI - Outpatient treatment of infective endocarditis. PMID- 11869263 TI - Antibiotic prophylaxis for infective endocarditis. PMID- 11869264 TI - Epidemiology of Chlamydia pneumoniae. AB - Chlamydia pneumoniae is the most commonly occurring intracellular bacterial pathogen. It is frequently involved in respiratory tract infections and to a lesser degree in extrapulmonary diseases. According to seroepidemiologic surveys, C. pneumoniae infection seems to be both endemic and epidemic. Such studies indicate that C. pneumoniae infection is widespread, with frequent reinfection during a lifetime. In Western countries the highest rate of new infections occurs between the ages of 5 and 15. The antibody prevalence worldwide is higher in adult males than in females. Currently available data suggest that C. pneumoniae is primarily transmitted from human to human without any animal reservoir. Transmission seems to be inefficient, although household outbreaks with high transmission rates are reported. Most reports rank C. pneumoniae among the three most common etiologic agents of community-acquired pneumonia, with an incidence ranging from 6% to 25%, and generally presenting a mild and, in some cases, self limiting clinical course. Recent reports also indicate a possible role for C. pneumoniae in severe forms of community-acquired pneumonia and in respiratory infections in immunocompromised patients. C. pneumoniae infection has also been implicated in the pathogenesis of asthma in both adults and children. The hypothesis that C. pneumoniae infection could lead to asthma is based on clinical studies and on the evidence of specific IgE production, direct epithelial damage, induction of T-cell immunopathologic diseases, and vascular smooth cell infection. Chronic C. pneumoniae infection seems to be common in patients with chronic bronchitis whether exacerbated or not, and is characterized by a strong humoral immune response to this intracellular microorganism, which is present in the majority of patients with severe chronic bronchitis. More than 60% of subjects with chronic bronchitis have specific C. pneumoniae antibody titers, and the microorganism may be identified by culture or PCR in almost 40% of these patients. This pathogen has also been recently associated with atherosclerosis and coronary heart disease (CHD). Seroepidemiological evidence indicates that the majority of patients with CHD present an anti-C. pneumoniae antibody pattern consistent with chronic infection. Furthermore, C. pneumoniae has been detected in atherosclerotic coronary plaques by several methods, including immunocytochemistry, transmission electron microscopy and molecular biology techniques. Recently, we detected C. pneumoniae DNA in a high percentage (51%) of aortic aneurysm plaques. Moreover, our serologic data support the hypothesis that a chronic C. pneumoniae antibody pattern may be a possible risk marker for atherosclerosis. Recently, C. pneumoniae has been isolated by culture from the coronary artery of a patient with coronary atherosclerosis, providing direct evidence of the presence of viable organisms in atheromatous lesions. Moreover, we recently demonstrated an association between C. pneumoniae reinfection and acute myocardial infarction. PMID- 11869265 TI - Diagnosis of Chlamydia pneumoniae. AB - This paper considers the use of diagnostic techniques for Chlamydia pneumoniae in both acute and chronic infections, together with the detection of upper respiratory tract carriage. It is concluded that such diagnosis is still unsatisfactory, although techniques for acute infection are more reliable than those for chronic conditions. Reliable and inexpensive methods, particularly for the diagnosis of chronic C. pneumoniae infection, are urgently needed, if only because of the recent demonstration of the beneficial effects of antibiotic treatment in patients with atherosclerosis carrying markers for the presence of this organism. PMID- 11869266 TI - Role of intracellular pathogens in respiratory tract infections. AB - Recent epidemiologic studies have shown that in patients with acute respiratory tract infection, intracellular pathogens, and particularly Mycoplasma pneumoniae and Chlamydia pneumoniae, might be involved more frequently than was previously assumed. In addition, newer investigations, such as specific culture or polymerase chain reaction (PCR), when systematically performed in patients with asthma, chronic obstructive pulmonary disease and even cystic fibrosis, frequently detect Mycoplasma pneumoniae or Chlamydia pneumoniae in those individuals. The significance of such detection (chronic bronchial infection?) and the role that these intracellular pathogens might play in the natural history of chronic respiratory diseases remain to be evaluated. PMID- 11869267 TI - Preclinical models for Chlamydia pneumoniae and cardiovascular disease: hypercholesterolemic mice. AB - Seroepidemiologic studies have shown an association of Chlamydia pneumoniae antibody with atherosclerosis. Compelling additional evidence has accumulated, in that the organism has been found within atherosclerotic lesions throughout the arterial tree by multiple methods. C. pneumoniae has also been isolated from coronary and carotid atheromatous plaques. Although these studies support a potential role for C. pneumoniae in atherogenesis, confirmation of a causal relationship requires the use of animal models and intervention studies. We have focused on the evaluation of mouse models to address the hypothesis that, following upper respiratory tract infection, lung macrophages are infected, disseminate to the aorta, and alter the onset or progression of atherogenesis. ApoE-deficient knock-out and C57BL/6J mice were used. The apoE-deficient mouse develops atherosclerotic lesions spontaneously on a regular diet in a time- and age-dependent manner. This knock-out strain was developed on the background of the C57BL/6J mouse, which only develops atherosclerosis on a high-fat/high cholesterol diet. To investigate whether infected macrophages constitute a vehicle for dissemination of C. pneumoniae in vivo, mice were inoculated intranasally or intraperitoneally. The organism was detected in harvested alveolar and peritoneal macrophages at all time points following intranasal and intraperitoneal inoculations, respectively, and in peripheral blood mononuclear cells following inoculation by both routes. In another experiment, alveolar and peritoneal macrophages from intranasally and intraperitoneally inoculated mice were adoptively transferred by intraperitoneal injection to uninfected mice. Subsequently, C. pneumoniae was detected in lung, spleen, abdominal lymph nodes and/or thymus of recipient mice. In control experiments, UV-inactivated C. pneumoniae DNA was not detected in alveolar or peritoneal macrophages beyond 5 min after inoculation in vivo. These cumulative results demonstrate that C. pneumoniae infects macrophages in vivo and that macrophages can serve as a vehicle for dissemination to other sites. To answer the question of whether the organism disseminates to and persists in the aorta, 8-week-old mice were infected intranasally. Following single or multiple inoculations in apoE-deficient mice, C. pneumoniae was detected in the lung and aorta for 20 weeks postinfection. In contrast, in C57BL/6J mice, the organism did not persist in the aorta following a single intranasal inoculation, but could be detected up to 7 weeks postinfection in multiply inoculated mice. Significantly, in apoE-deficient mice with developed atherosclerotic lesions, the organism was found in foam cells within the lesions by immunocytochemical staining. These studies show that persistent C. pneumoniae infection occurs in atherosclerotic lesions in the aorta in the apoE-deficient knock-out mouse model. Infection of the aorta also occurred in C57BL/6J mice but was more transient. Both models should be useful in studying the pathogenic role of C. pneumoniae in atherogenesis and for determining if therapy prevents dissemination of infection. To this end, we have evaluated the susceptibility of C. pneumoniae to roxithromycin, a new macrolide, in vitro in cell culture and in vivo in the apoE-deficient mouse model. In vitro results compare favorably with other new macrolides, quinolines, and tetracyline. Preliminary studies in the apoE model of persistent infection suggest that the organism is detected less frequently in the lung and aorta by PCR following treatment. These promising preliminary studies warrant further investigation. PMID- 11869269 TI - Discussion and Conclusion. PMID- 11869268 TI - Link between intracellular pathogens and cardiovascular diseases. AB - Atherosclerosis is a multifactorial disease with an important inflammatory component. Inflammatory cells such as T-lymphocytes, macrophages and monocytes play a key role not only in its origin but also during plaque rupture. In addition, it has been suggested that infection could be the cause of atherosclerosis. Serologic studies involving polymerase chain reaction, immunocytochemistry and electron microscopy support the hypothesis of an association between Chlamydia pneumoniae and atherosclerosis. This intracellular pathogen is able to replicate in macrophages, vascular cells and smooth muscle cells. C. pneumoniae can then release inflammatory cytokines which contribute to instability of the plaque. C. pneumoniae may be recovered through cell culture from coronary atherosclerotic plaques and has a strong association with one allele, of the major histocompatibility system among patients with acute coronary syndromes. Other intracellular pathogens that have been implicated in atherosclerosis include cytomegalovirus, Mycoplasma pneumoniae and Helicobacter pylori. To test the hypothesis that C. pneumoniae plays a role in the instability of plaque, we conducted a prospective clinical trial, using the macrolide antibiotic roxithromycin in 202 patients. The preliminary results show a statistically significant reduction in the incidence of acute coronary events at 30 days. PMID- 11869270 TI - Do we need fourth-generation cephalosporins? PMID- 11869271 TI - Clinical activity of cefepime in severe infections. PMID- 11869272 TI - Patterns and modes of beta-lactam resistance in Pseudomonas aeruginosa. PMID- 11869273 TI - Antibiotic policy: a tool for controlling resistance of hospital pathogens. AB - Multiresistant Gram-negative bacilli, including strains of Klebsiella pneumoniae, Enterobacter spp, Acinetobacter baumannii and Pseudomonas aeruginosa, resistant to broad spectrum beta-lactams, aminoglycosides and fluoroquinolones, are recovered at increasing frequency from patients suffering from nosocomial infections, particularly from those receiving intensive care. The emergence and spread of resistant pathogens to endemic and epidemic levels has frequently been related in time and place to the intensive use of antibiotics to which these microorganisms have developed resistance, notably third generation cephalosporins and fluoroquinolones. Recent investigations have indicated that the prevalence of resistance can be reduced by scheduled changes of empiric treatment regimens, involving discontinuation of intensively prescribed drugs and substitution with newly introduced antibiotics of another class to which the prevalent resistant strains remain susceptible. Among these drugs, penicillins---beta-lactamase inhibitor combinations, 'fourth generation' cephalosporins and, where little used previously, fluoroquinolones, have been introduced successfully in high risk units where ceftazidime-resistant strains of K.pneumoniae, Enterobacter and Citrobacter spp or glycopeptide-resistant enterococci had become highly prevalent. However, these studies do not demonstrate a direct causal relationship between changes in prescribing practices and ecological improvements, because their observational design cannot be controlled. In most studies, several important factors influencing the dynamics of resistance were not monitored and the relative contribution of decreased emergence versus control of cross transmission to the improved susceptibility rates is not clear. We propose that additional long-term studies are required to better track the ecological impact and to determine the optimal modalities of programmed changes of antibiotic prescribing as an antibiotic resistance prevention or control strategy. PMID- 11869274 TI - In vitro selective concentrations of cefepime and ceftazidime for AmpC beta lactamase hyperproducer Enterobacter cloacae variants. AB - This study aimed to compare the selective concentrations of cefepime and ceftazidime on Enterobacter cloacae. A mixed culture of a wild-type ceftazidime/cefepime-susceptible (4x107 CFU/mL) strain and an ampC derepressed Enterobacter cloacae (105 CFU/mL) strain (relative proportions 99.75% and 0.25%) was challenged for 4 h with different antibiotic concentrations of ceftazidime and cefepime (0.03--4096 mg/L), and then transferred to drug-free medium. The proportion of wild-type versus derepressed population was evaluated after 24 h. Ceftazidime and cefepime selected the derepressed variant at concentrations ranging from 1 to 4096 and from 0.12 to 16 mg/L respectively. These results suggest that serum concentrations attainable with a 2 g/8 h cefepime dosage may be able to suppress the emergence of derepressed ampC mutants. PMID- 11869275 TI - Strategies for selecting antibiotics in intensive care units. AB - Crowding of severely ill patients in intensive care units has led worldwide to important increases in nosocomial (ICU-related) infections. Moreover, the nature of these hospital-acquired infections is shifting towards Gram-positive microorganisms, yeast and Gram-negative rods, possessing important resistance genes (e.g. extended spectrum beta-lactamases and inducible Enterobacteriaceae). Ceftazidime and aztreonam are loosing their activity against the Gram-negative microorganisms. The fourth generation cephalosporins have an intrinsic high activity against the inducible Enterobacteriaceae. On our Hematology and Intensive Care units, the introduction of cefepime for nosocomial infections led to a remarkable drop in the number of Enterobacter isolates combined with important decreases in Enterobacter resistance towards several antibiotics. PMID- 11869276 TI - Conclusion: Fourth-generation cephalosporins: new hopes and new duties. PMID- 11869277 TI - A multinational workshop on drug-resistant pathogens of the respiratory tract: prevalence, risk factors and intervention strategies. PMID- 11869278 TI - Treatment options for resistant pneumococcal infections. PMID- 11869280 TI - Development and testing of Streptococcus pneumoniae conjugate vaccines. PMID- 11869279 TI - Antibiotic usage and the incidence of resistance. PMID- 11869281 TI - Carriage of resistant pneumococci by children in southern Israel and impact of conjugate vaccines on carriage. PMID- 11869282 TI - Antibiotic use and pneumococcal resistance to penicillin: the French experience. PMID- 11869283 TI - Modification of prescribers' behavior: the Icelandic approach. PMID- 11869284 TI - Role of communicable disease control measures in affecting the spread of resistant pneumococci: the Swedish model. PMID- 11869286 TI - The challenge of multiresistant Streptococcus pneumoniae: international initiatives in day-care centers and the use of molecular epidemiologic techniques. PMID- 11869285 TI - Carriage and antibiotic resistance of respiratory pathogens and molecular epidemiology of antibiotic-resistant Streptococcus pneumoniae colonizing children in day-care centers in Lisbon: the Portuguese day-care center initiative. PMID- 11869287 TI - The bone morphogenetic protein antagonist Noggin is regulated by Sox9 during endochondral differentiation. AB - Noggin has been described to be capable of binding bone morphogenetic proteins (BMP) and inhibiting BMP signaling by preventing the interactions of BMP with their receptors. Noggin expression during endochondral differentiation was analyzed to elucidate its potential role during chondrogenesis. Throughout mouse development, Noggin was expressed abundantly in the chondrocytic lineage as early as collagen type II RNA was detectable. In addition, a strong correlation was detected between Noggin expression and the expression profile of Sox9 during chondrogenesis. Sox9 (known to play an important role during chondrogenesis) and Noggin expression were investigated in the pluripotent mesenchymal cell line C3H10T1/2, stimulated by BMP-2. BMP-2 caused significant upregulation of Sox9 and Noggin expression in these cells. The upregulation of Noggin could be inhibited by introducing antisense oligonucleotides against Sox9 mRNA into the cells. Using mouse limb bud cultures, the role of Sox9 and Noggin during endochondral tissue differentiation was further studied. Treatment of cultures with Sox9 antisense oligonucleotides and/or Noggin protein caused significant alterations in limb morphogenesis and endochondral development. The data suggest that the transcriptional control of Noggin by Sox9 is a potent regulatory mechanism in chondrocyte differentiation. PMID- 11869288 TI - Absorption, transportation and digestion of egg white in quail embryos. AB - The present study was done to reveal how egg white is taken up by embryonic tissues, the pathway through which egg white is transported, and the location where it is digested during the development of the quail Coturnix japonica. Antiserum against quail ovalbumin was raised in rabbit and used as a probe. By immunoelectron microscopy, the uptake of ovalbumin on a small scale by receptor mediated endocytosis was observed in the ectodermal cells of the yolk sac on days four to seven of incubation. The uptake of egg white on a large scale by fluid phase endocytosis took place in the cells generally referred to collectively as the 'albumen sac'. The ovalbumin was transported through the albumen sac into the extraembryonic cavity during days eight to 10, and then into the amniotic cavity through the amnion approximately on day 10. Ovalbumin was present in the intestinal lumen on days 11 and 14, but it was not digested in the intestinal epithelial cells. The ovalbumin was detected in the yolk of embryos after day 10. Immunoblot testing, as well as a fluoroimmunoassay, revealed that the location where the amount of ovalbumin was highest changed chronologically from the extraembryonic cavity on day 10 to the amniotic cavity on day 11, the intestinal lumen on day 12 and then to the yolk on day 13. Several low molecular proteins which cross-reacted with the antiserum were observed in the extracts of the yolk. The reaction producing these proteins depended on low pH (approximately 3.0) and was inhibited by pepstatin A. The ovotransferrin was similarly digested. These results indicate that egg white is, for the most part, transported through the albumen sac to the yolk via the extraembryonic cavity, the amniotic cavity, and the intestinal lumen, and is digested in the yolk by aspartic proteinases. PMID- 11869289 TI - Possible role of the 38 kDa protein, lacking in the gastrula-arrested Xenopus mutant, in gastrulation. AB - An acidic, 38 kDa protein that is present in Xenopus wild-type embryos has been previously shown to be lacking in gastrula-arrested mutant embryos. To gain understanding of the role of this protein, its spatio-temporal distribution and involvement in gastrulation was investigated using the monoclonal antibody (9D10) against it. The protein was prominent in the cortical cytoplasm of cells facing the outside in the animal hemisphere of embryos until the gastrula stage, and in ciliated epithelial cells of embryos at stages later than the late neurula. When the 9D10 antibody was injected into fertilized wild-type eggs, they cleaved normally, but most of them had arrested development, always at the early stage of gastrulation, as in the mutant embryos. In contrast, the majority of the control antibody-injected eggs gastrulated normally and developed further. Cytoskeletal F actin, which was mainly observed in the area beneath the plasma membrane facing the outside of the epithelial layer of not only the dorsal involuting marginal zone but also the dorsal, vegetal cell mass of the control antibody-injected embryos at the early gastrula stage, was scarcely recognized in the corresponding area of the 9D10 antibody-injected embryos. It is likely that the paucity of the F-actin caused by the 9D10 antibody inhibition of the 38 kDa protein might lead to a failure of cell movement in gastrulation, resulting in developmental arrest. PMID- 11869290 TI - Matrix metalloproteinases regulate mesonephric cell migration in developing XY gonads which correlates with the inhibition of tissue inhibitor of metalloproteinase-3 by Sry. AB - In the mouse, the sex determining gene Sry, on the Y chromosome, controls testis differentiation during embryogenesis. Following Sry expression, indifferent XY gonads increase their size relative to XX gonads and form cord-like structures with the adjacent mesonephros, providing XY gonad somatic cells. This mesonephric cell migration is known to depend on Sry, but the molecular mechanism of mesonephric cell migration remains unknown. In this study, it was shown that cells expressing Sry induced proliferation of mesonephric cells migrating into male gonads, and inhibited expression of the tissue inhibitor of metalloproteinases (TIMP)-3 gene, which is the endogenous inhibitor of matrix metalloproteinases (MMP). In addition, the mesonephric cell migration was blocked by a chemically synthesized inhibitor of MMP in a gonad/mesonephros organ co culture system with enhanced green fluorescent protein transgenic embryos. The findings indicate that MMP may play a critical role in mesonephric cell migration, and the function of MMP may be regulated by a Sry-TIMP-3 cascade. These findings are an important clue for the elucidation of testicular formation in developing gonads. PMID- 11869291 TI - Role of syndecan in the elongation of postoral arms in sea urchin larvae. AB - Ac-SYN is the core protein of a cell surface proteoglycan of the sea urchin Anthocidaris crassispina. To examine the functions of Ac-SYN, embryos were cultured in the presence of affinity-purified antibody against Ac-SYN. At the late pluteus stage, severe inhibition of elongation of the postoral arms was seen in treated embryos compared with control embryos. Blastocoeleic microinjection of the antibody did not affect morphogenesis. The relationship between the number of cells in the postoral arms and the length of the postoral rods was investigated in normal embryos. This showed that postoral arm elongation has two phases: the first phase accompanies the increase in cell numbers while the second does not. The syndecan antibody inhibited the increase in cell numbers in the postoral arms. Furthermore, in the treated embryos, cell numbers continued to increase normally until 31 h post fertilization (hpf), while cell division stopped after 31 hpf. These results suggest that Ac-SYN participates in postoral arm formation via cell division in sea urchin embryos. PMID- 11869292 TI - Xenopus Brachyury regulates mesodermal expression of Zic3, a gene controlling left-right asymmetry. AB - The Brachyury gene has a critical role in the formation of posterior mesoderm and notochord in vertebrate development. A recent study showed that Brachyury is also responsible for the formation of the left-right (L-R) axis in mouse and zebrafish. However, the role of Brachyury in L-R axis specification is still elusive. Here, it is demonstrated that Brachyury is involved in L-R specification of the Xenopus laevis embryo and regulates expression of Zic3, which controls the L-R specification process. Overexpression of Xenopus Brachyury (Xbra) and dominant-negative type Xbra (Xbra-EnR) altered the orientation of heart and gut looping, concomitant with disturbed laterality of nodal-related 1 (Xnr1) and Pitx2 expression, both of which are normally expressed in the left lateral plate mesoderm. Furthermore, activation of inducible type Xbra (Xbra-GR) induces Zic3 expression within 20 min. These results suggest that a role of Brachyury in L-R specification may be the direct regulation of Zic3 expression. PMID- 11869293 TI - Role of the thrombopoietin (TPO)/Mpl system: c-Mpl-like molecule/TPO signaling enhances early hematopoiesis in Xenopus laevis. AB - Multiple organs are induced in the primitive embryonic ectoderm excised from blastula stage Xenopus laevis embryos, under the strict control of mesoderm inducing factors. This in vitro system is useful for exploring the mechanisms of development. In this study, the function of thrombopoietin (TPO)/c-Mpl signaling in the development of hematopoietic cells was investigated. An optimal hematopoietic cell induction system was established to evaluate the influence of growth factors on hematopoiesis. It was found that exogenous TPO enhanced hematopoiesis in explants induced by activin and bone morphogenetic protein (BMP) 4 and increased the number of both erythrocytes and leukocytes in a dose dependent manner. Addition of anti-c-Mpl antibody completely inhibited the expansion of hematopoietic cells stimulated by TPO, and the antibody specifically recognized blood-like cells. These results demonstrate that TPO acts on hematopoietic progenitors induced in explants and the c-Mpl-like molecule in Xenopus mediates the cellular function of TPO. We also found that forced expression of TPO in embryos promoted hematopoiesis in the ventral blood island and the dorsal-- lateral plate mesoderm. These results suggest that hematopoietic stem and progenitor cells are regulated by TPO/c-Mpl signaling from when they appear in their ontogeny. They also suggest that TPO/c-Mpl signaling play a crucial role in the formation of hematopoietic cells in Xenopus. PMID- 11869294 TI - Region-specific gastrointestinal Hox code during murine embryonal gut development. AB - Hox genes encode transcription factors, and they are involved in the specification of each body part along the anteroposterior (AP) body axis during embryogenesis. To clarify AP pattern formation of the digestive tract, the expression patterns of Hox genes belonging to paralogous groups 4 and 5, and parts of groups 6 and 7, were systematically examined by whole-mount and section in situ hybridization. The Hox gene expression pattern of paralogous groups 4-9 in the developing gut at 12.5 days post-coitum was fully examined. All HoxA and HoxB genes in paralogous groups 4-8 were expressed in the stomach, in contrast to the HoxC and HoxD genes. In the midgut region, all Hox cluster genes showed colinear expression within each cluster, yielding the Hox code; the more 3' located genes were expressed more rostrally and the 5' group genes more caudally. The colinear expression of HoxA and HoxB cluster genes started from the duodenum, that of HoxC cluster genes started from the jejunum, and HoxD cluster genes were expressed in the caudal part of the midgut, ileum and cecum. In the hindgut region, HoxD cluster genes and Abd-B family genes were expressed. Thus, a different Hox code seems to exist in each subdomain of developing gut (foregut, midgut and hindgut). The visceral mesoderm restricted expression also suggested that the Hox code primarily functions in mesenchymal specification, and then leads to the regional differentiation of gut subdomains as the result of epithelial-mesenchymal interactions. PMID- 11869295 TI - In the multiheaded strain (mh-1) of Hydra magnipapillata the ectodermal epithelial cells are responsible for the formation of additional heads and the endodermal epithelial cells for the reduced ability to regenerate a foot. AB - Hydra consist of three self-renewing cell lineages: the ectodermal epithelial, endodermal epithelial and interstitial cell lineages. The role of these cell lineages in head formation and foot regeneration in Hydra magnipapillata was studied by comparing the multiheaded strain mh-1 with the wild-type. Adult polyps of this strain show a reduced ability to regenerate a foot in the apical body half several days before additional heads are formed there. Cell lineage chimeras were produced, and it was found that in mh-1, the ectodermal epithelial cell lineage is responsible for the formation of additional heads, whereas the endodermal epithelial cell lineage and, to a lesser extent, the derivatives of the interstitial cell lineage, are responsible for the reduced ability of foot regeneration. PMID- 11869297 TI - Patient-assessed health outcome measures for diabetes: a structured review. AB - AIMS: To identify available disease-specific measures of health-related quality of life (HRQL) for diabetes and to review evidence for the reliability, validity and responsiveness of instruments. METHODS: Systematic searches were used to identify instruments. Instruments were assessed against predefined inclusion and exclusion criteria. Letters were sent to authors requesting details of further instrument evaluation. Information relating to instrument content, patients, reliability, validity and responsiveness to change was extracted from published papers. RESULTS: The search produced 252 references. Nine instruments met the inclusion criteria: Appraisal of Diabetes Scale (ADS), Audit of Diabetes Dependent Quality of Life (ADDQoL), Diabetes Health Profile (DHP-1, DHP-18), Diabetes Impact Measurement Scales (DIMS), Diabetes Quality of Life Measure (DQOL), Diabetes-Specific Quality of Life Scale (DSQOLS), Questionnaire on Stress in Diabetic Patients-Revised (QSD-R), Diabetes-39 (D-39) and Well-being Enquiry for Diabetics (WED). The shortest instrument (ADS) has seven items and the longest (WED) has 50 items. The ADS and ADDQoL are single-index measures. The seven multidimensional instruments have dimensions covering psychological well being and social functioning but vary in the remainder of their content. The DHP 1 and DSQOLS are specific to Type 1 diabetes patients. The DHP-18 is specific to Type 2 diabetes patients. The DIMS and DQOL have weaker evidence for reliability and internal construct validity. Patients contributed to the content of the ADDQoL, DHP-1/18, DQOL, DSQOLS, D-39, QSD-R and WED. The authors of the ADDQoL, DHP-1/18, DQOL, DSQOLS gave explicit consideration to content validity. The construct validity of instruments was assessed through comparisons with instruments measuring related constructs and clinical and sociodemographic variables. None of the instruments has been formally assessed for responsiveness to changes in health. CONCLUSIONS: Five of the diabetes-specific instruments have good evidence for reliability and internal and external construct validity: the ADDQoL, DHP-1/18, DSQOLS, D-39 and QSD-R. Instrument content should be assessed for relevance before application. The instruments should be evaluated concurrently for validity and responsiveness to important changes in health. PMID- 11869298 TI - The influence of the polymorphism in apolipoprotein B codon 2488 on insulin and lipid levels in a Danish twin population. AB - AIMS: The apolipoprotein B codon 2488 polymorphism has been associated with the metabolism of lipoproteins in subjects with Type 2 diabetes. However, no data are available on the influence of the polymorphism on insulin or glucose metabolism. This study examines the impact of the polymorphism on parameters associated with the insulin resistance syndrome in Danish twins. METHODS: The effect of the polymorphism on lipid, glucose and insulin measures was studied in 548 same sex twins aged 55-74 years. RESULTS: The codon 2488 polymorphism influenced fasting triglyceride levels, as well as insulin, as measured at 120 min in an oral glucose tolerance test. Subjects with the genotype T2488T had 14% higher triglyceride levels (P = 0.02) and 31% higher insulin levels (P = 0.004) than subjects with genotype C2488C. In twins discordant for genotype, the T-allele was associated with higher levels of triglyceride (P = 0.04) and insulin (P = 0.02) and lower levels of HDL-cholesterol (P = 0.04). CONCLUSION: The T-allele of the codon 2488 polymorphism influenced parameters related to the insulin resistance syndrome, i.e. increased levels of insulin, increased levels of triglyceride and decreased levels of HDL. As the polymorphism is silent, these effects must be mediated through linkage to other polymorphisms in apolipoprotein B or other genes on chromosome 2. PMID- 11869299 TI - Elevation of von Willebrand factor in plasma in diabetic patients with neuropathic foot ulceration. AB - AIMS: The present study examines the relationship between markers of endothelial dysfunction and diabetic neuropathy or consequent neuropathic foot ulceration in patients with Type 2 diabetes mellitus. METHODS: We studied 65 Type 2 diabetic patients including 25 diabetic patients without neuropathy, 27 with neuropathy but no history of foot ulceration, and 13 with neuropathic ulceration. Plasma concentrations of von Willebrand factor (vWF) and soluble thrombomodulin (TM), measures of endothelial dysfunction, were determined by enzyme immunoassays. We performed various tests quantifying aspects of diabetic neuropathy including vibration perception threshold (VPT; for sensory neuropathy), coefficient of variation of R-R intervals (CVR-R; for cardiac autonomic neuropathy), and cold induced vasodilation in the great toe for peripheral sympathetic neuropathy. RESULTS: CVR-R and cold-induced vasodilation were significantly diminished in patients with neuropathic foot ulceration compared with patients with neuropathy but no history of foot ulceration. Plasma vWF concentrations were positively correlated with VPT and cold-induced vasodilation test, and were inversely correlated with CVR-R. Multivariate analysis disclosed that VPT and percentage vasodilation were independent factors for plasma vWF. Plasma vWF was significantly elevated in patients with foot ulceration compared with patients without neuropathy or those with neuropathy but not foot ulceration. However, plasma TM concentrations did not differ between the three groups. CONCLUSIONS: Diabetic patients with neuropathic foot ulceration had severe impairment of cardiac autonomic and peripheral sympathetic nerves. Elevation of vWF in plasma was associated with neuropathic foot ulceration, linking endothelial dysfunction to foot ulceration. PMID- 11869300 TI - What is the prevalence of visual impairment in the general and diabetic populations: are there ethnic and gender differences? AB - AIMS: To estimate the prevalence of blindness and partial sight in the general and diabetic populations and to measure the impact of ethnicity and gender on these rates. METHODS: Data on visual impairment in the population to be studied was obtained from the Royal Leicestershire Rutland and Wycliffe Society for the Blind (RLRWSB). Using the validated technique of capture-mark-recapture (CMR), a census day was set and data from four sources collected: hospital admissions data, GP returns, wards, and mass media. Computer cross-linkage to Leicestershire Health Authority data ensured patients were alive and resident in Leicestershire at the census date. Estimates of diabetes, ethnicity and gender prevalence were calculated using the 1991 census. RESULTS: CMR calculations revealed that there was under ascertainment of visual impairment in the RLRWSB data. The CMR calculations with direct age standardization estimate the prevalence of blindness and partial sight in the general and diabetic populations to be 51.8/10 000 (CI 50.6-53.0) and 148.8/10 000 (CI 122.1-175.5), respectively. In the diabetic population there was a higher proportion of visually impaired females than males (P < 0.05), with no overall ethnic differences; however, rates were lower than expected in female Indo-Asians. In the general population there was a higher proportion of visually impaired Indo-Asian people (P < 0.01). CONCLUSIONS: Our study has shown the prevalence of visual impairment in both the general and diabetic populations and has demonstrated both ethnic and gender differences. CMR allows the calculation of prevalence of blindness and partial sight from existing data. When comparing the prevalence of visual impairment, whether from diabetes or other causes, it is important to know the gender and ethnicity of the 'at risk' population, as well as the rates in the background population. PMID- 11869302 TI - Risk of diabetes in siblings of index cases with Type 2 diabetes: implications for genetic studies. AB - AIMS: The goal was to estimate the sibling recurrence-risk ratio for Type 2 diabetes in families with diabetes occurring in middle age. Because diabetes aetiology involves environmental exposures and genetic susceptibility, we sought to identify determinants of the recurrence risk. METHODS: We surveyed patients diagnosed at ages 35-59 years (n = 563) to obtain information on the occurrence of diabetes in their relatives, particularly siblings (n = 1675). Age-specific prevalences of diabetes in the US population were used for comparison. RESULTS: The overall sibling recurrence-risk ratio for diabetes was low, about 1.8 in the Joslin families and even lower in three other studies that were reanalysed for comparison. In all studies, the diabetes risk in siblings of index cases without a history of diabetes in a parent was similar to that in the general population, suggesting that genetic factors contributed to the occurrence of diabetes in only a minority of these siblings. The fact that recurrence-risk ratios were elevated only in families with one or two diabetic parents indicates that susceptibility to Type 2 diabetes is transmitted primarily through an affected parent. In addition, the sibling recurrence-risk ratios were elevated even further in families with diabetes in both a parent and grandparent of the index case, and in siblings of non-obese index cases (percent ideal body weight < 120%). CONCLUSIONS: The selection of families with non-obese index cases and vertical transmission of diabetes through three generations may improve the success of efforts to map susceptibility genes for Type 2 diabetes. PMID- 11869301 TI - Amputations and diabetes: a case-control study. AB - AIMS: Data concerning the relative risk of amputations in diabetic patients compared with the general population are scarce. Therefore, we carried out a case control study to quantify the relationship between diabetes and amputations. METHODS: In 20 hospitals in seven German cities and counties we obtained complete lists of non-traumatic lower limb amputations performed in 1990 and 1991. CONTROLS were selected from patients of the same surgical departments operated on in the same years. We drew a random sample of patients with procedures not likely to be associated with diabetes. Diabetic status was determined from patients' records in both cases and controls. We calculated age- and sex-specific and, using logistic regression, adjusted odds ratios (OR) and attributable risks. RESULTS: N = 2400, mean age 61.7 (SD 16.3) years. CASES: n = 729; 486 (66.7%) of them had diabetes. CONTROLS: n = 1671; 127 (7.6%) of them had diabetes. Adjusted OR: 18.2 (confidence interval (CI) 14.2-23.6). Adjusted attributable risk among exposed (ARE): 0.95 (CI 0.93-0.96). Adjusted population attributable risk (PAR): 0.62 (CI 0.57-0.66). CONCLUSIONS: This study has demonstrated a strong association between the risk of amputation and diabetes. The odds ratios and attributable risks for diabetic individuals are higher in the younger than in the older age groups. Population attributable risks are great. We conclude that the reduction of amputations in the general population will be achieved by improving foot care in people with diabetes. PMID- 11869303 TI - The short insulin tolerance test lacks validity in adolescents with Type 1 diabetes. AB - AIMS: The short insulin tolerance test (SITT) has been found to be a simple and valid method for determining insulin sensitivity in healthy adults and patients with Type 2 diabetes. In this study we evaluated the reproducibility and validity of SITT in 16 adolescents with Type 1 diabetes. METHODS: Thirteen patients underwent two SITT and eight patients were examined with both SITT and a euglycaemic hyperinsulinaemic clamp. At the SITT insulin sensitivity was measured from the slope of arterialized blood glucose concentrations determined for 16 min after an intravenous bolus injection of short-acting insulin, 0.1 U/kg body weight, and expressed as glucose disappearance rate (KITT). RESULTS: There was a significant correlation between the insulin sensitivity estimations made at the two SITT (r = 0.73, P = 0.003). The reproducibility was low, however, with a coefficient of variation of 38.7%. KITT showed a strong inverse correlation to the fasting blood glucose concentration (r = -0.74, P < 0.0001). We found no correlation between insulin sensitivity measured by SITT and that measured by the euglycaemic clamp. CONCLUSIONS: We conclude that the short insulin tolerance test cannot be used in adolescent patients with Type 1 diabetes for a simple estimation of insulin sensitivity. PMID- 11869304 TI - The effect of cigarette smoking on soluble adhesion molecules in middle-aged patients with Type 2 diabetes mellitus. AB - AIMS: To investigate the effect of smoking on soluble adhesion molecules in middle-aged diabetic patients. METHODS: One hundred out-patients with Type 2 diabetes and 100 age- and sex-matched non-diabetic subjects without clinical macrovascular disease were selected. Soluble serum levels of adhesion molecules were analysed using enzyme immunoassay. Carotid atherosclerosis was assessed using an ultrasound system. RESULTS: When compared with non-diabetic subjects, soluble vascular cell adhesion molecule-1 (sVCAM-1), soluble intercellular adhesion molecule-1 (sICAM-1), and sE-selectin were found at significantly high levels in diabetic patients and significantly higher levels of sICAM-1, sE selectin, and sP-selectin were observed in current smokers than never-smokers among diabetic or non-diabetic subjects, respectively. The combined, but not enhanced, effects of diabetes mellitus and smoking were observed in sICAM-1 and sE-selectin levels. Additionally, levels of sICAM-1 (P < 0.05) and sE-selectin (P < 0.01), but not sP-selectin, were high in ex-smokers when compared with never smokers among diabetic patients. Diabetic smokers were also found to have marked carotid atherosclerosis, which was related to increased levels of sICAM-1. CONCLUSIONS: Our present study shows that levels of adhesion molecules were higher in diabetic smokers than diabetic non-smokers or non-diabetic smokers, and that cessation after chronic smoking did not restore the levels of sICAM-1 and sE selectin, though sP-selectin levels were restored. These data suggest a possible mechanism for accelerated atherosclerosis induced by smoking in patients with diabetes. PMID- 11869305 TI - Erythropoietin response to hypoxia in patients with diabetic autonomic neuropathy and non-diabetic chronic renal failure. AB - AIMS: An erythropoietin (EPO)-deficient anaemia is recognized in Type 1 diabetic patients with early nephropathy and symptomatic autonomic neuropathy (DN). The aim of this study was to determine whether the EPO response to hypoxia was deficient in order to clarify the mechanisms involved in this process. METHODS: Five Type 1 diabetic patients DN (age 39 (28-48) years (mean (range))) with EPO deficient anaemia (haemoglobin, Hb 10.6 (9.5-12.0) g/dl, EPO 5.0 (3.2-6.5) IU/l) and early diabetic nephropathy (persistent proteinuria 1161.6 (130-2835) mg/day, serum creatinine 97.6 (63-123) micromol/l)) were compared with nine normal subjects (age 31 (24-39) years, Hb 13.4 (11.8-15.7) g/dl, EPO 7.6 (5.6-10.3) IU/l) and four patients with non-diabetic advanced chronic renal failure RF (proteinuria 2157.5 (571-4578) mg/day, serum creatinine 490.2 (406-659) micromol/l, Hb 10.3 (9.0-11.3) g/dl, EPO 4.6 (2.9-8.5) IU/l). The subjects were exposed to 6 h of hypoxia (inspired oxygen 11.6-12.6%) by breathing a gas mixture via a hood. Hourly serum EPO levels were measured. RESULTS: All groups showed a rise in EPO production after 2 h. The diabetic DN group achieved a similar maximal response to the normal subjects at 6 h (EPO 17.3 +/-5.4 vs. 17.8 +/-7.9 IU/l). The renal failure patients mounted an EPO response to hypoxia but at lower EPO levels. CONCLUSIONS: Although the DN patients have inappropriately low EPO levels for the severity of their anaemia, they can mount an appropriate EPO response to moderate hypoxia. The mechanism underlying the EPO-deficient anaemia present in some diabetic patients remains unclear. PMID- 11869306 TI - The effect of subclinical hypothyroidism on metabolic control in children and adolescents with Type 1 diabetes mellitus. AB - AIMS: Associated autoimmune phenomena might influence metabolic control in children and adolescents with Type 1 diabetes mellitus. A retrospective case control study was performed in order to explore the effect of subclinical hypothyroidism on metabolic control in Type 1 diabetes mellitus. PATIENTS AND METHODS: For this purpose each patient with Type 1 diabetes and subclinical hypothyroidism (cases) was matched for age, duration of disease and, if possible, for sex, with two to three diabetic patients without hypothyroidism (controls). Parameters of metabolic control such as HbA1c, total insulin requirement and frequency of symptomatic hypoglycaemia were retrieved for 12, 6 and 3 months before and after diagnosis of hypothyroidism. RESULTS: Thirteen patients (two male/11 female) patients were diagnosed with subclinical hypothyroidism and were matched with 31 controls (nine male/22 female). There was no difference (mean and range) in terms of age (11.9 years (4.4-18.1) vs. 11.7 years (3.5-18.1), P = 0.9) and duration of disease (5.1 years (1.2-10.5) vs. 4.38 years (0.9-10.8), P = 0.6) between the two groups. There was no difference in HbA1c and total insulin requirement between the two groups at any time point of assessment (anova P = 0.8 and P = 0.1, respectively). Patients with hypothyroidism had significantly more symptomatic hypoglycaemic episodes during the 12 months before diagnosis (anova P = 0.05), increasing progressively during this time period and reaching a peak at time 0 (5.5+/-0.4 vs. 1.6+/-0.1 episodes/month, P = 0.01). No difference could be detected within 6 months of starting substitution therapy (2.4+/-0.2 vs. 1.6+/ 0.1 episodes/week, P = 0.8). CONCLUSIONS: These data suggest that subclinical hypothyroidism is associated with an increased risk of symptomatic hypoglycaemia. The prompt introduction of substitution therapy is recommended as it reduces its frequency. PMID- 11869307 TI - An integrated mobile foot and retinal screening programme for people with Type 2 diabetes managed in primary care. AB - AIMS: To undertake a vascular and neurological assessment on the feet of all patients with Type 2 diabetes managed solely in local primary care. METHODS: A mobile screening podiatrist (working with an existing mobile retinal screening programme) screened a selected population of 4022 patients with Type 2 diabetes managed solely in 82 general practices. Doppler pressure assessments of peripheral vasculature, bioesthesiometer and monofilament assessment of peripheral neuropathy. RESULTS: This service was administratively simple to set up and integrated well with the retinal screening service and secondary care foot clinic, and was valued by the practices. Disease prevalences were 1.04% for foot ulceration, 19% for peripheral vascular disease and up to 29% for peripheral neuropathy. CONCLUSIONS: This programme screens all patients with known diabetes managed solely in primary care within a district and describes foot morbidity and allows risk stratification. This pattern of service could be a useful template for discussing the diabetes National Service Framework with primary care groups. PMID- 11869308 TI - Diabetes care in cats and dogs. PMID- 11869309 TI - Prevalence of gestational diabetes mellitus in Hong Kong based on the 1998 WHO criteria. PMID- 11869310 TI - ADA/WHO diabetes diagnostic criteria controversy---the Chennai chapter. PMID- 11869312 TI - A problem with the diagnosis of diabetes mellitus based on fasting plasma glucose. PMID- 11869313 TI - Predicting CHD risk in patients with diabetes. PMID- 11869314 TI - Colon interposition following esophagectomy. PMID- 11869315 TI - Gastroplasty: yes or no to gastric drainage procedure. PMID- 11869316 TI - Evolution of DNA ploidy during squamous cell carcinogenesis in the esophagus. AB - Image and flow cytometry was used to study the nuclear DNA content (ploidy) during the squamous cell carcinogenesis in the esophagus. The present retrospective study comprised 26 surgical specimens of squamous cell carcinomas (SCC) in patients who underwent surgery alone at the Department of Surgery in CHUV Hospital in Lausanne, between January 1992 and December 1999. We analyzed 53 healthy tissues, 43 tumors, and six lymph node metastases. Diploid DNA histogram patterns were observed in all non-pathologic tissues analyzed, either distant or proximal to the lesion. Aneuploidy was observed in 30 (70%) of 43 lesions; 20 (62.5%) of 32 early squamous-cell carcinomas; and 10 (91%) of 11 advanced carcinomas. In patients with various tumor stages or with multicentric synchronous or metachronous tumors, DNA content was not different among different tumor stages. Four of six lymph node metastases had the same DNA content as the primary tumor. In four patients, discordance between image and flow cytometry analysis was observed for malignant lesions only. Ploidy status was not statistically associated with the differentiation of the tumor, but it was associated with the stage of tumor (P < 0.001). These findings suggest that early malignant changes in the esophagus are already associated with alteration in DNA content, and aneuploidy tends to correlate with progression to invasive SCC. This cell kinetic information could help clinicians in selecting the optimal treatment for the individual patient. PMID- 11869317 TI - p53 protein in esophageal mucosa of individuals at high risk of squamous cell carcinoma of the esophagus. AB - Squamous cell carcinoma of the esophagus (SCCE) is diagnosed late and carries a poor prognosis. Biomarkers such as p53 protein expression may be present in the esophageal mucosa long before esophageal symptoms or lesions appear and may point toward early diagnosis. Asymptomatic subjects at high risk for SCEE (consumption of more than 80 g of ethanol and 10 cigarettes/day for at least 10 years) underwent upper gastrointestinal endoscopy with biopsies of the esophageal mucosa, and expression of p53 protein was compared with conventional histologic findings. In 182 subjects studied, p53 protein was expressed in a stepwise fashion according to the severity of the histologic findings: normal mucosa (12/103 or 11.7%), mild chronic esophagitis (6/43 or 14%), moderate chronic esophagitis (4/18 or 22.2%), severe chronic esophagitis (1/3 or 33.3%), low-grade dysplasia (4/11 or 36.4%), high-grade dysplasia (2/2 or 100%), and squamous cell carcinoma (2/2 or 100%) (P=0.00025). The odds ratio and confidence intervals were calculated by logistic regression, with multivariate adjustment for potentially confounding variables. The risk for p53 expression was twofold for moderate and severe chronic esophagitis and 10-fold for dysplasia and cancer (P=0.001). p53 protein was expressed not only in cancerous lesions, high-grade and low-grade dysplasia, as expected, but also in mucosa considered normal or with chronic esophagitis using conventional histology. Smokers and alcohol drinkers with normal mucosa or chronic esophagitis that express p53 protein may represent an unrecognized subgroup of individuals that may benefit from surveillance. Follow up studies of these asymptomatic subjects and molecular analysis of the p53 gene are needed to clarify this point. PMID- 11869318 TI - Lymph node metastasis along the recurrent nerve chain is an indication for cervical lymph node dissection in thoracic esophageal cancer. AB - This study examined whether recurrent nerve chain node metastasis serves as an indicative factor for cervical lymph node dissection in thoracic esophageal cancer. The association of recurrent nerve chain lymph node metastasis and cervical node metastasis was analyzed for 91 patients with thoracic esophageal cancer who had undergone three-field lymph node dissection. In patients with upper thoracic esophageal cancer, the incidence of cervical lymph node metastasis was similar regardless of recurrent nerve chain node metastasis. On the other hand, in patients with middle or lower esophageal cancer, the incidence was significantly higher in recurrent nerve-positive (16/31, 51.6%) than in recurrent nerve-negative (5/43, 11.6%) patients. The prognosis of patients with recurrent nerve chain node metastasis was significantly better in the three-field dissection group than in the two-field dissection group, while in patients with no recurrent nerve chain node metastasis, survival was similar between the two groups. In conclusion, cervical lymphadenectomy can be omitted for recurrent nerve chain node-negative patients with middle and lower thoracic esophageal cancer. PMID- 11869319 TI - Chemoradiotherapy followed by surgery for thoracic esophageal cancer potentially or actually involving adjacent organs. AB - The objective of this study was to evaluate the therapeutic usefulness of chemoradiotherapy (CRT) followed by surgery in patients with clinically T4 (cT4) esophageal cancer involving adjacent organs such as the trachea, main bronchi, and large vessels. Thirty-seven patients with cT4 squamous cell carcinoma of the thoracic esophagus were enrolled in this study. The CRT regimen comprised cisplatin (70 mg/m2) on day 1, 5-fluorouracil (700 mg/m2) on days 1-4 and external irradiation (200 cGy/day, total 30 Gy) on either days 8-26 (sequential schedule, n=15) or days 1-19 (concurrent schedule, n022). Two courses of CRT were given. The results of CRT were complete response in nine patients, partial response in 19, no change in three (minor response in two), and progressive disease in six patients. The median response duration in all responders was 172 days (range: 56-2469, n=19). After CRT, 13 patients received surgery. In 12 of these patients, tumors were completely resected. Histopathologic examination of the resected specimen revealed a discrepancy between clinical response and histopathologic effect. The median duration of survival and the 1-, 2- and 5-year survival rates were 304 days (84-3155), 45%, 35% and 23% in all patients, respectively, 866 days (190-3155), 83%, 83% and 57% in the 13 patients whose tumors were resected, and 187 days (84--2630), 25%, 5% and 5% in the 24 patients whose tumors were not resected. Grade 3 toxicity, especially hematological reactions, was noted in 13.5% (5/37) of the patients. There was one toxicity related death (sepsis). A good outcome may be obtained with CRT, followed by surgery when feasible. However, CRT can cause toxic reactions, and close monitoring of patients is required. PMID- 11869320 TI - Bi-modality (chemo-radiation) versus tri-modality (chemo-radiation followed by surgery) treatment for carcinoma of the esophagus. AB - The purpose of the study was to investigate the difference in overall survival in patients with localized carcinoma of esophagus treated using chemo-radiation (bi modality, BM) or chemo-radiation followed by surgery (tri-modality, TM). From 1981 to 1999, 65 patients were identified who had localized carcinoma of the esophagus treated with either concurrent chemo-radiation (BM, n=22) or concurrent chemo-radiation followed by surgery (TM, n=43) at the University of Texas Medical Branch at Galveston. All 65 patients received concurrent chemotherapy and external beam radiation. Radiation was delivered by linear accelerators (greater than-or-equal 6 MV), except in one patient who had part of his treatment given by a Co-60 machine. Chemotherapy consisted of 5-fluorouracil and cisplatin plus minus vinblastine under different regimens. Median follow-up time was 10 months (range=1-195 months) for all patients. Of the 14 patients still alive, the median follow-up time was 32 months (range=2-192 months). No difference in overall survival was detected between the two treatment groups, BM vs. TM (P=0.394) despite a selection bias favoring the TM group. Five-year survival rates of the BM and TM groups were 17% and 18%, respectively; 10-year survival rates were 17% and 12%, respectively. The presence of significant past medical history (P=0.017) and a complete pathologic response in the TM group (P < 0.001) were significant independent predictors of survival. We did not find any difference in survival between chemo-radiation or chemo-radiation followed by surgery in patients with localized carcinoma of the esophagus. Use of biologic markers and functional imaging should be explored in order to segregate patients with different tumor biology for treatment using different treatment strategies. PMID- 11869321 TI - Comparative study of self-expandable metallic stent and bypass surgery for inoperable esophageal cancer. AB - We evaluated the effect of two different therapies for 23 patients with inoperable esophageal cancer, of whom 13 patients underwent placement of a metallic stent and 10 received bypass surgery. We newly defined the improved level of food intake and the effective rate based on the grade of the patient's status in order to evaluate objectively. Although the effect of stent placement was significantly better than that of bypass surgery, there were no significant differences in the improved level of food intake and the effective rate between the two groups. Four out of 10 patients in the bypass group had suture insufficiency, resulting in no oral intake and a fatal outcome. In addition, the ratio of survival period of ability to ingest orally and in-hospital mortality was significantly favorable in the stent group compared with the bypass group. In conclusion, stent placement should be the first choice for inoperable malignant stenosis or esophageal fistula. PMID- 11869322 TI - Handsewn or stapled esophagogastric anastomoses after esophagectomy for cancer: meta-analysis of randomized controlled trials. AB - Gastric transposition with esophagogastric anastomosis is a common method of reconstruction after esophagectomy for cancer. The anastomosis can be fashioned using a handsewn or stapled technique. The choice of anastomotic technique is often debated but there is little evidence to support the use of one method over the other. We performed a meta-analysis of randomized controlled trials (RCTs) to determine the effect of esophagogastric anastomotic method (handsewn or circular stapled) on patient outcomes. Medline and manual searches were done (completed independently and in duplicate) to identify all published RCTs that addressed the issue of handsewn or stapled esophagogastric anastomosis after esophagectomy for cancer. The selection process was inclusive; no trials were excluded. Trial validity assessment was done and a trial quality score was assigned. Major outcomes for quantitative data synthesis included operative mortality, anastomotic leaks, anastomotic strictures, cardiac morbidity, and pulmonary morbidity. A random-effects model was used and relative risk was the principal measure of effect. Systematic qualitative review was used for other outcomes such as duration of operation and time to complete the anastomosis. Data on cancer survival were not available in the RCTs. Five RCTs were selected with quality scores ranging from 2 to 3 (5-point Jadad scale). Selection and validity agreement was strong. Relative risk (95% confidence interval, CI; P-value), expressed as handsewn vs. stapled (treatment vs. control), was 0.45 (0.20, 1.00; P=0.05) for operative mortality, 0.79 (0.44, 1.42; P=0.43) for anastomotic leaks, 0.60 (0.27, 1.33; P=0.21) for anastomotic strictures, 0.99 (0.55, 1.77; P=0.97) for cardiac morbidity, and 0.93 (0.63, 1.37; P=0.72) for pulmonary morbidity. Data synthesized from existing RCTs show that handsewn and circular stapled esophagogastric anastomotic techniques give similar results for anastomotic outcomes, such as leaks and strictures. The stapled anastomotic method appears to increase operative mortality (P=0.05). Although it is difficult to explain this finding, it should not be dismissed. Several hypotheses are discussed. PMID- 11869323 TI - Human cadavers as an experimental model for esophageal surgery. AB - The use of cadavers in experimental esophageal surgery is reviewed. Items useful to cadaveric studies such as post-mortem changes, biosafety, ethics, and legislation are discussed. Tactics used in minimally invasive procedures (thoracoscopy and laparoscopy) are shown. Cadaveric use in studies concerning esophagectomy, gastroesophageal reflux disease, esophageal atresia, Boerhaave's syndrome, and Mallory-Weiss tears are discussed. It is concluded that human bodies represent a good but underused model for esophageal surgery. PMID- 11869324 TI - Prognostic value of laparoscopic ultrasound in patients with gastro-esophageal cancer. AB - Forty-four patients with gastro-esophageal tumors regarded as resectable by conventional staging underwent laparoscopic ultrasonography (LUS). Following LUS, seven were found to be irresectable and were managed by palliative therapies. Thirty-seven patients proceeded to surgical exploration and 36 were resected (R0 80%, R1 11%, and R2 9%). All patients were reviewed until death or for a minimum of 24 months. Patients undergoing resection had a 62% 1-year survival (median 17 months; confidence intervals, CI 6-28). LUS defined nodal status indicated a trend toward prolonged survival in the node-negative group, median 22 months (CI 5-39), compared with 13 months (CI 6-20) in the node-positive group. Disease-free survival was greater in LUS node-negative patients at 29 months (CI 23-35) compared with node-positive patients at 13 months (CI 5-21) P=0.0083. LUS staging allows prediction of the likelihood of recurrence of gastro-esophageal malignancies. This may prove useful for the appropriate allocation of patients to primary and adjuvant therapies. PMID- 11869325 TI - Randomized controlled trial of intrasphincteric botulinum toxin A injection versus balloon dilatation in treatment of achalasia cardia. AB - As the few randomized controlled trials available in the literature comparing botulinum toxin (BT) injection with established endoscopic treatment of achalasia cardia, i.e. pneumatic dilatation, showed conflicting results, we conducted a prospective randomized trial. Seventeen consecutive patients with achalasia cardia diagnosed during a period between December 1997 and February 2000 were randomized into two treatment groups [pneumatic dilatation by Rigiflex dilator (n=10), BT injection by sclerotherapy needle into four quadrants of lower esophageal sphincter (LES) (n=7) 80 units in five cases, 60 units in two cases] after dysphagia grading, endoscopy, barium esophagogram, and manometry, all of which were repeated 1 week after treatment. Patients were followed up clinically for 35.2+/-14 weeks. Chi-squares, Wilcoxon rank-sum test, Kaplan-Meier method and log-rank tests were used for statistical analysis. After 1 week, 6/7 (86%) BT treated vs. 8/10 (80%) dilatation-treated patients improved (P=NS). There was no difference in LES pressure and maximum esophageal diameter in the barium esophagogram in the two groups before therapy. Both therapies resulted in significant reduction in LES pressure. The cumulative dysphagia-free state using the Kaplan-Meier method decreased progressively in BT-treated compared with dilatation-treated patients (P=0.027). Two patients with tortuous megaesophagus, one of whom had failed dilatation complicated by perforation previously, improved after BT. One other patient in whom pneumatic dilatation had previously failed improved in a similar manner. BT is as good as pneumatic dilatation in achieving an initial improvement in dysphagia of achalasia cardia. It is also effective in patients with tortuous megaesophagus and previous failed pneumatic dilatation. However, dysphagia often recurs during 1-year follow up. PMID- 11869326 TI - Lower esophageal sphincter pressure in idiopathic achalasia and Chagas disease related achalasia. AB - The most important etiologies of achalasia are idiopathic and related to Chagas' disease. The lower esophageal sphincter pressure (LESP) in idiopathic achalasia (Id Ach) is higher compared with a healthy group, but there are different reports in Chagasic achalasia (Ch Ach). We compared the LESP of patients with both forms of achalasia and a control group. The LESP of 213 achalasia patients without previous treatment and 32 healthy volunteers were assessed. In 126 patients, the etiology could be demonstrated using serologic tests (Id Ach, 94 and Ch Ach, 32). The LESP of 213 patients was 31.86+/-14.18 mmHg and in the control group was 17.92+/-7.03 mmHg (P < 0.0001). The LESP in Id Ach and Ch Ach was 33.28+/-13.63 mmHg and 23.5+/-12.09 mmHg (P < 0.0001), respectively. Only the Id Ach group achieved statistical difference in relation to the control group (P < 0.0001). In conclusion, the LESP of Id Ach patients was higher than in Ch Ach patients and the control group, but there was no LESP difference between the Ch Ach and control groups. PMID- 11869328 TI - Primary malignant melanoma of the esophagus treated by endoscopic ablation and interferon therapy. AB - Primary malignant melanoma of the esophagus is a rare illness accounting for 0.1 0.2% of malignant diseases of the esophagus; however, the incidence of the disease appears to be rising. The average survival time is between 10 and 15 months. The authors describe the 25 month follow up of a patient with primary malignant melanoma of the esophagus which was treated with endoscopic ablation followed by interferon therapy. No other focus was found and the patient is undergoing regular endoscopic check-ups, currently without any problems. PMID- 11869327 TI - Gastro-esophageal reflux disease confined to the sphincter. AB - It has been shown previously that patients with gastro-esophageal reflux disease (GERD) do not always have increased esophageal acid exposure on 24 h pH monitoring. The recent recognition of carditis as a sensitive marker for GERD raises the possibility for patients with mild disease to have normal esophageal acid exposure but inflamed cardiac mucosa on biopsies of the cardia, which may be an early sign of GERD. To test this hypothesis, 171 consecutive patients evaluated for symptoms of GERD and no increased esophageal acid exposure, Barrett's esophagus or erosive esophagitis were divided into those with and without carditis. Esophageal acid exposure and lower esophageal sphincter (LES) characteristics were compared between the two groups. Comparisons were done using the Mann-Whitney U-test for non-parametric data. There were 82 patients with histologic evidence of carditis and 89 patients without carditis. Patients with carditis had a more deteriorated sphincter, determined by overall and abdominal length and resting pressure, and significantly higher esophageal acid exposure (P < 0.05). Patients with symptoms of GERD and histologic evidence of carditis may have early or mild reflux disease, which is confined to the sphincter. PMID- 11869329 TI - Primary malignant melanoma of the esophagus treated by esophagectomy and systemic chemotherapy. AB - We describe herein a case of asymptomatic primary malignant melanoma of the esophagus. A 65-year-old man presented with a 4-cm filling defect in the middle third of the esophagus on a routine barium swallow. Subtotal esophagectomy accompanied by lymph node dissection was performed through a right thoracotomy. Postoperatively, the patient received five cycles of systemic chemotherapy with dacarbazine (DTIC), nimustine hydrochloride (ACNU), and vincristine (VCR) (DAV therapy), but ultimately died of generalized metastatic disease 15 months after surgery. Malignant melanoma of the esophagus has an extremely poor prognosis despite various therapeutic efforts. PMID- 11869331 TI - Direct esophageal metastasis from a distant primary tumor is a submucosal process: a review of six cases. AB - Malignant esophageal stricture secondary to invasion from a tumor arising in a contiguous organ is a relatively rare finding; even more uncommon is a direct metastasis to the esophagus from a distant primary carcinoma. We present six cases, the largest current series, of esophageal strictures secondary to metastases from a separate primary cancer. We reviewed the records of 20 patients treated at Virginia Mason Medical Center between 1972 and 2000 with a diagnosis of malignant esophageal stricture secondary to an extraesophageal primary carcinoma. Patients whose stricture appeared to be secondary to esophageal invasion or compression from a contiguous tumor or lymph nodes were excluded. The remaining six patients who had metastases to the esophagus itself were reviewed with respect to the nature of the primary tumor, presentation, radiologic and endoscopic findings, and treatment. Among the 20 patients reviewed, 14 were excluded owing to either contiguous involvement from a nearby primary malignancy, regional nodal involvement, or complications of external beam radiation treatment. Six patients were considered to have direct metastasis to the esophagus from distant primary malignancies. The mean age of these patients was 72 years (range 68-74). Two of the primary lesions were lung carcinoma, while four primaries were breast cancers. The average time interval from the diagnosis of a primary tumor to esophageal involvement was 7 years in patients with breast cancer and 5 months in patients with lung cancer. Three patients were palliated with endoscopic dilation and stent placement. The other three patients have died secondary to upper gastrointestinal bleeding. Metastatic cancer to the esophagus is a rare occurrence. The process is usually submucosal and can be difficult to diagnose. The diagnosis should be considered when a patient presents with malignant dysphagia and has a background of distant carcinoma. PMID- 11869330 TI - Simultaneous leiomyosarcoma and squamous cell carcinoma of the esophagus: report of a new case. AB - Leiomyosarcoma of the esophagus is an uncommon neoplasm of mesenchymal origin. The simultaneous occurrence of leiomyosarcoma and squamous cell carcinoma in the esophagus as separate tumors is a very rare event. We present a case leiomyosarcoma associated with squamous cell carcinoma that was diagnosed after surgical resection. The clinical, radiologic, endoscopic and pathologic findings of this rare entity, and four additional cases previously reported in the literature are reviewed. PMID- 11869332 TI - Emergency re-operation for postoperative hemorrhage following partial esophagectomy for carcinoma of the esophagus and cardia of the stomach. AB - The clinical records of 12 cases of emergency re-operation for management of postoperative hemorrhage (POH) following partial esophagectomy and esophago gastrostomy or colonic interposition for a group of 3690 cases of carcinoma of the esophagus (CE) and cardia of the stomach in this institute between August 1954 and April 2001 were studied. There were 10 survivors and two deaths, giving a mortality rate of 16.6% (2/12). It was concluded that an emergency re-operation was a cardinal surgical procedure to save the patient's life if a serious POH had developed. The POH could be prevented by a careful, conscientious and strict hemostasis during the primary operation. The criteria for the diagnosis are presented. PMID- 11869333 TI - Covered metallic stent treatment of a patient with spontaneous rupture of the esophagus. AB - A patient with a potentially fatal condition as a result of esophago-pneumo broncho fistula was successfully treated with the insertion of a self-expanded covered metallic stent. Severe regurgitation resulted in the removal of the stent 3 months after insertion. Stricture after removal of the stent required pneumatic balloon dilation. The use of a self-expanded covered metallic stent is effective for the treatment of spontaneous esophageal rupture; however, early removal of the stent is recommended. PMID- 11869334 TI - Squamous cell carcinoma arising from a congenital duplication cyst of the esophagus in a young adult. AB - Squamous cell carcinomas arising from the congenital anomalies in the esophagus are rare. One such case of an 18-year-old man, with an associated history of ventricular septal defect, who developed an epithelial malignancy within the duplication cyst extending to involve the lower third of esophagus is presented here. He responded well to radical treatment using concurrent chemo-irradiation, and continues to be free of disease after a follow-up of 14 months. PMID- 11869335 TI - Association of multiple granular cell tumors and squamous carcinoma of the esophagus: case report and review of the literature. AB - This report describes the case of a man who underwent subtotal esophagectomy for the concomitant presence of a multifocal esophageal squamous carcinoma and a granular cell tumor (GCT); he had been previously affected by another metachronous esophageal GCT excised endoscopically. This is the sixth case described in the literature detailing other cases of a combination of malignancies involving additional organs. We emphasize the need for a prolonged surveillance of patients with multiple GCTs in order to promptly recognize the possibility of associated neoplasms. PMID- 11869336 TI - Simultaneous surgical management of iatrogenic pharyngeal perforation and distal esophageal malignancy. AB - Instrumental perforation of the pharynx with distal obstruction is a complex problem. A fistula is not likely to close in the presence of distal obstruction. The stenotic lesion needs to be treated in addition to the perforation. We report a 83-year-old female patient who underwent three-stage total esophagectomy and right cervical pharyngo-gastric anastomosis for iatrogenic pharyngeal perforation and distal esophageal malignancy. The radical surgical approach has the advantage of treating the immediate crisis due to perforation and also treating the stricture for which the esophagoscopy was originally performed. PMID- 11869338 TI - Broncho-gastric fistula complicating transthoracic esophagectomy. AB - Tracheo-esophageal and broncho-esophageal fistulas are recognized complications in advanced esophageal cancers. The common complications seen after transthoracic esophagectomy include pulmonary complications, anastomotic leaks, gastric stasis, and anastomotic strictures. A broncho-gastric fistula is a very rare complication of transthoracic esophagectomy and has been reported anecdotally in the literature. We report a patient who underwent a transthoracic esophagectomy and developed a broncho-gastric fistula caused by a staple from the stomach tube suture line eroding into the right bronchus. Diagnosis was made by fiberoptic bronchoscopy. The patient was successfully treated with re-exploratory thoracotomy and closure of the fistula. PMID- 11869339 TI - Inflammatory pseudotumor of the esophagus. AB - Inflammatory pseudotumors of the esophagus are uncommon. They pose diagnostic and therapeutic dilemmas, especially when located in the cervical esophagus. History and physical examination are rarely contributory. Routine radiologic investigations including barium swallow and computed tomography only raise the suspicion of a benign esophageal neoplasm. Esophagoscopy and biopsy do not provide a definite diagnosis, as these 'tumors' are frequently submucosal, unless they enlarge sufficiently to cause mucosal ulcerations. Endoscopic ultrasonography may accurately localize the tumor but is not diagnostic. Conservative surgical resection or debulking would be both diagnostic and therapeutic. Steroids, cyclophosphamide and low-dose radiotherapy may at best be considered second-line therapy. One such case of inflammatory pseudotumor of the cervical esophagus is presented and the relevant literature is reviewed. Our patient could not be diagnosed on preoperative investigations and required an esophagotomy with frozen section. We debulked the mass and the patient is asymptomatic 6 months after surgery. PMID- 11869337 TI - Bilateral bronchial stenting and esophageal intubation for advanced bronchial carcinoma with esophageal invasion. AB - Successive bilateral bronchial stenting (Dumon type) and minimally invasive pull through esophageal intubation for accompanying malignant bronchial and esophageal involvement was undertaken. External radiation and afterloading brachytherapy for localized endobronchial overgrowth was used. A 13-month survival was achieved using mainly out-patient facilities. During such esophageal intubation, bronchoscopic control is mandatory. Extended complex palliation was obtained using this combined treatment, even in the high-risk stage of advanced tracheobronchial carcinoma with associated esophageal stricture. PMID- 11869340 TI - Laparoscopic excision of esophageal leiomyoma through an anterior esophagotomy. AB - Esophageal leiomyoma are usually intramural and the most common minimally invasive approach is thoracoscopic excision leaving the mucosa intact. We report the case of a 58-year-old woman who underwent laparoscopic excision of a pedunculated esophageal leiomyoma. PMID- 11869341 TI - Seminal fluid-mediated fitness traits in Drosophila. AB - The seminal fluid of male Drosophila contains a cocktail of proteins that have striking effects on male and female fitness. In D. melanogaster, seminal fluid proteins affect female receptivity, ovulation, oogenesis, sperm storage, sperm competition and mating plug formation. In addition, the seminal fluid contains antibacterial peptides and protease inhibitors. Some seminal fluid-encoding genes also show high rates of evolutionary change, exhibiting both significant between species divergence and within-species polymorphism. Seminal fluid protein genes are expressed only in males, begging the question of how and why the reproductive processes of females are influenced by males. In this review I address these issues by bringing together evidence for the function, evolution, diversification, and maintenance of variation in, seminal fluid-mediated traits. PMID- 11869342 TI - Multiplicity of infection and the evolution of hybrid incompatibility in segmented viruses. AB - Some viral genomes are divided into segments. When multiple viruses infect a single cell, progeny form by reassorted mixtures of genomic segments. Hybrid incompatibilities arise when a progeny virus has incompatible segments from different parental viruses. Hybrid incompatibility has been observed in influenza and in the multiparticle plant virus Dianthovirus. Hybrid incompatibility provides an opportunity to study rates of viral evolution, divergence and speciation, and the extent of epistatic interactions among components of the viral genome. This paper presents mathematical and computer simulation models to study hybrid incompatibility between diverging strains. The models identify multiplicity of infection as a key factor. When many viral particles infect each host cell, the effective ploidy of the genetic system is high. High ploidy dilutes the contribution of each locus to the phenotype, weakening the selective intensity on each locus. Weaker selection on variant alleles allows the population to maintain greater genetic diversity and to be more easily perturbed by stochastic fluctuations. Greater diversity and stochastic fluctuations explore more widely the space of epistatic interactions, causing more frequent shifts among favoured combinations of alleles. Variable ploidy of viral genetics differs from standard Mendelian genetics. PMID- 11869344 TI - Analysis of segregation data from selfed progeny of allopolyploids. AB - This paper discusses the inference of parental genotype based on segregation data from selfed progeny of allopolyploids when there is incomplete information about genotypes and when alleles are codominant or null. The distinct alleles that are present in a genotype are assumed to be known, but not the frequency with which they occur. These assumptions may be appropriate when genotypes are deduced from DNA or protein banding patterns on electrophoretic gels. A computer program, SELF, is described that can generate all possible parental genotypes and rank them on the basis of their agreement with the progeny data. The program caters for tetraploids, hexaploids and octoploids. The methods are illustrated using data from a study of the inheritance of isoenzymes in selfed progeny of octoploid strawberry cultivars. PMID- 11869343 TI - Genetic basis for co-adaptive gene complexes in rice (Oryza sativa L.) landraces. AB - One hundred and twenty-two AFLP markers were mapped using an IR64 x Azucena rice doubled-haploid (DH) population. The distribution of these mapped markers was monitored across a set of 48 diverse landraces of rice. Strong statistical associations were observed between 960 of the 7381 possible pairs of markers across the diverse material. These 960 strongly associated pairs of markers mapped to the same chromosomes in only 111 cases. The remaining 849 pairs were the result of association between markers found on different chromosomes. More than 21% of these genetically unlinked but strongly associated markers are not randomly distributed across the genome but instead occupy blocks of DNA on different rice chromosomes. Amongst associated blocks, there has clearly been maintenance of combinations of marker alleles across very diverse germplasm. Analyses have also revealed that markers are found in association with performance for each of four quantitative traits in both the diverse landrace material and a DH mapping population. It is proposed that the present data provide strong evidence for the co-adaptation of geographically distinct landraces and that this has resulted over time in the maintenance of 'adaptive gene complexes' involving agronomically important quantitative traits. PMID- 11869346 TI - Fitness and microsatellite diversity estimates were not correlated in two outbred anuran populations. AB - Larval fitness traits were investigated in two anuran species (Bufo calamita and Rana temporaria) under controlled laboratory conditions, and allelic diversity measured in the same individuals at five and seven microsatellite loci, respectively. For both species there were significant differences among sibships in larval growth and development rates, and in some cases also in microsatellite heterozygosity and mean d2 (a measure of diversity based on differences in allele sizes). However, there were no significant correlations between any of the fitness and genetic measures either among all individuals, among sibships or among individuals within sibships. Under the conditions and within the statistical power of the study there was no relationship between fitness-related quantitative trait variation and that seen at presumed-neutral microsatellite loci in these outbred populations. PMID- 11869345 TI - Evolution of transcript structure and base composition of rDNA expansion segment D3 in ticks. AB - Four to thirty-two copies of the rDNA 28S gene expansion segment D3 and flanking H14 stem were sequenced in six species of ticks (Ixodes: Ixodidae: Acari). Sequence match among species varied from 66% to 97%. Sequence length averaged 130 bases in I. persulcatus across eight Eurasian sites and averaged 186 bases in five other species across 19 Eurasian and North American sites. The difference in length represents one or more deletions totalling about 60 bases that correspond to stems S3 or S4 of the folded transcript. The typical transcript conformation was observed as one possible low energy structure in the five species of longer D3. The structure entails a basal loop with four stem/loop structures, S1-S4 (moving 5' to 3') atop stem H14. A secondary structure lacking S4 but possessing all other putative standard features of the D3 transcript is possible with the shorter I. persulcatus sequences. Interspecific sequence differences occur at higher frequency in loops and bulges vs. complementary pairing regions of stems. Insertion/deletion events (indels) and base substitutions accounted equally for sequence differences. Indels are flanked by similar sequences, suggesting that they occur by slippage during replication. The D3 of Ixodes species is composed of a degenerate set of subrepeats. Thus, unequal exchange among subrepeats may have caused the reduction in length of the I. persulcatus D3. Compensatory base substitution and compensatory insertion/deletion events are indicated by the failure of mutations to affect secondary structure. Transversions accounted for 64% of sequence differences and were biased toward the gain of G and U and the loss of A and C. This bias could re-establish intramolecular base pairing when disrupted by insertions or deletions that shift one side of a stem relative to the other. The distribution of sequence differences, biased substitution, and conservation of transcript conformation in D3 suggest selective constraint. PMID- 11869347 TI - Genetic differentiation between mallee and tree forms in the Eucalyptus loxophleba complex. AB - The Eucalyptus loxophleba complex comprises two tree taxa (E. loxophleba ssp. loxophleba and ssp. supralaevis), two mallee taxa (E. loxophleba ssp. lissophloia and ssp. gratiae), and three rare mallee species, of which one, E. blaxellii, is included in this study. The genetic diversity and phylogenetic relationships between the taxa, particularly in relation to habit, were assessed using anonymous nuclear RFLP loci. The level of diversity in the taxa was high and similar to that detected in other eucalypt species. The populations showed low differentiation at both the subspecies and the species levels. Phylogenetic relationships showed some genetic separation between the tree and mallee habit but no separation of the two taxa within the tree habit or within the mallee habit. The genetic analysis does not support the recognition of E. gratiae as a separate species. The geographically restricted E. blaxellii showed similar levels of diversity to populations of the other widespread taxa of E. loxophleba. PMID- 11869348 TI - Fecundity and offspring ploidy in matings among diploid, triploid and tetraploid Chamerion angustifolium (Onagraceae): consequences for tetraploid establishment. AB - Models of polyploid evolution indicate that tetraploids are more likely to establish within diploid populations when they are formed recurrently through the union of unreduced (n=2n) gametes. To account for the coexistence of diploids and tetraploids in populations of Chamerion angustifolium, diploid, triploid and tetraploid plants were crossed in all possible combinations and fecundity and ploidy using flow cytometry of the resulting progeny were measured. Combined with previous data on cytotype fitness, these data were used in a simulation to examine the impact of unreduced gametes on tetraploid evolution. Seed set per fruit was highest in 2x x 2x crosses (69%), intermediate in 4x x 4x, 2x x 4x and 3x x 4x crosses (range, 11-35%) and lowest in 3x x 2x and 3x x 3x crosses (range, 1-10%). Offspring were diploid (94%) or triploid (6%) in 2x x 2x crosses, diploid (17.5%), triploid (56%) or tetraploid (26.5%) in 3x x 2x crosses, and triploid (53%) and tetraploid (44%) in all others (4x x 4x, 3x x 4x, 2x x 4x), indicating that some gametes are unreduced, particularly in triploids. Forty-two percent of offspring, from three different crosses, had DNA contents greater than tetraploids. Computer simulations based on these results showed that unreduced gamete formation via triploids in C. angustifolium can promote the coexistence of diploids and tetraploids, but, due to law triploid fitness, is insufficient to overcome tetraploid minority disadvantage. PMID- 11869349 TI - (TTAGGG)n telomeric sequence in selachian chromosomes. AB - The distribution of telomeric sequence (TTAGGG)n in the genomes of Chondrichthian species at different stages of evolution was investigated both by DNA genomic hybridization, and by fluorescence in-situ hybridization (FISH) of metaphase chromosomes. The sequence is highly conserved in all the species examined. FISH revealed a label uptake only by the telomeres in Raja asterias. However, in Torpedo ocellata, we revealed pericentromeric and interstitial sequence localization on some chromosome pairs in addition to the hybridization signal on telomeres. These findings confirm that the karyotype evolution of these fish began by Robertsonian fusion. PMID- 11869350 TI - Genetic control of self-incompatibility in Anagallis monelli (Primulaceae: Myrsinaceae). AB - The genetic control of self-incompatibility (SI) was studied in the Mediterranean short-lived perennial species Anagallis monelli (Primulaceae: Myrsinaceae). Arrays of siblings, including families derived from reciprocal crosses, were cross-pollinated in full diallels, and compatibility groups were assesssed from a census of fruit-set. Two, three and four intercompatible and intraincompatible groups were found. These crossing relationships fit the model for gametophytic SI controlled by a single polymorphic gene locus in families derived from parents with one or no S alleles in common (two vs. four compatibility groups), whilst one genotype was presumed to be missing in the small families that showed only three compatibility groups. PMID- 11869351 TI - Heterosis in two closely related species of earthworm (Eisenia fetida and E. andrei). AB - The importance of heterosis, and in particular allozyme-associated heterosis, in natural populations remains unclear. Much of the scepticism that surrounds allozyme-associated heterosis comes from inconsistent and unreliable detection of the relationship. Thus, evaluating the genetic mechanisms that may cause allozyme associated heterosis in natural populations has proven difficult. The most prevalent hypotheses that have been put forward to explain the genetic basis of heterosis are the general dominance and the local overdominance hypotheses. A factorial crossing design was used to survey eight polymorphic allozyme loci in the parent and offspring generations of two species of earthworms in order to evaluate possible mechanisms of allozyme-associated heterosis for growth rate. Significant heritable variation for growth rate was detected only within a single cross. Allozyme-associated heterosis for growth rate was detected only within this cross. This relationship did not persist after the effects of interfamily variation were removed. These results indicate that simple heritability of a fitness-related trait may be necessary for predictive power and repeatability of allozyme-associated heterosis, and that the allozyme-associated heterosis detected in this study was the result of general dominant genetic effects. PMID- 11869357 TI - CD8+CD28- T cells: certainties and uncertainties of a prevalent human T-cell subset. AB - Human peripheral blood CD8+ T cells comprise cells that are in different states of differentiation and under the control of complex homeostatic processes. In a number of situations ranging from chronic inflammatory conditions and infectious diseases to ageing, immunodeficiency, iron overload and heavy alcohol intake, major phenotypic changes, usually associated with an increase in CD8+ T cells lacking CD28 expression, take place. CD8+CD28- T cells are characterized by a low proliferative capacity to conventional stimulation in vitro and by morphological and functional features of activated/memory T cells. Although the nature of the signals that give origin to this T-cell subset is uncertain, growing evidence argues for the existence of an interplay between epithelial cells, molecules with the MHC-class I fold and CD8+ T cells. The possibility that the generation of CD8+CD28- T cells is the combination of TCR/CD3zeta- and regulatory factor mediated signals as a result of the sensing of modifications of the internal environment is discussed. PMID- 11869358 TI - Experimental autoimmune encephalomyelitis induction in naive mice by dendritic cells presenting a self-peptide. AB - Self-reactive T cells escape deletion in the thymus and are found in the peripheral repertoire. Because bone-marrow-derived dendritic cells (BM-DC) are potent activators of antigen-specific T cells, these cells could theoretically activate self-reactive T cells leading to autoimmunity. We investigated whether BM-DC could induce the autoimmune disease experimental autoimmune encephalomyelitis (EAE). Our results show that transfer of BM-DC presenting a self-peptide from the myelin oligodendrocyte glycoprotein (MOG35-55) into naive mice induced EAE 7-14 days later. MOG35-55-specific T cells of the Th1 phenotype were present in the lymph nodes and spleens of mice that received live peptide pulsed BM-DC. Heat-killed or formaldehyde-fixed BM-DC presenting MOG35-55 could induce neither clinical signs of EAE nor a measurable T-cell response in vitro. These data show that live BM-DC presenting a self-antigen can induce the organ specific autoimmune disorder EAE in a non-transgenic system. Therefore, this new EAE model could be used as a more clinically relevant model for the human disease multiple sclerosis. These findings could also have implications for the use of DC immunotherapy in a clinical setting. PMID- 11869360 TI - Does recycling in germinal centres exist? AB - A general criterion is formulated to decide if the recycling of B cells exists in germinal centre (GC) reactions. The criterion is independent of the selection and affinity maturation process and based solely on total centroblast population arguments. An experimental test is proposed to verify whether the criterion is fulfilled. PMID- 11869359 TI - Route of administration of chimeric BPV1 VLP determines the character of the induced immune responses. AB - To examine the mucosal immune response to papillomavirus virus-like particles (PV VLP), mice were immunized with VLP intrarectally (i.r.), intravaginally (i.va.) or intramuscularly (i.m.) without adjuvant. PV-VLP were assembled with chimeric BPV-1 L1 proteins incorporating sequence from HIV-1 gp120, either the V3 loop or a shorter peptide incorporating a known CTL epitope (HIVP18I10). Antibody specific for BPV-1 VLP and P18 peptide was detected in serum following i.m., but not i.r. or i.va. immunization. Denatured VLP induced a much reduced immune response when compared with native VLP. Immune responses following mucosal administration of VLP were generally weaker than following systemic administration. VLP specific IgA was higher in intestine washes following i.r. than i.va. immunization, and higher in vaginal washes following i.m. than i.r. or i.va. immunization. No differences in specific antibody responses were seen between animals immunized with BPV-1 P18 VLP or with BPV-1 V3 VLP. Cytotoxic T lymphocyte precursors specific for the P18 CTL epitope were recovered from the spleen following i.m., i.va. or i.r. immunization with P18 VLP, and were similarly detected in Peyer's patches following i.m. or i.r. immunization. Thus, mucosal or systemic immunization with PV VLP induces mucosal CTL responses and this may be important for vaccines for mucosal infection with human papillomaviruses and for other viruses. PMID- 11869361 TI - Hepatoimmunology: a perspective. AB - Premises for the subspecialty of hepatoimmunology include the recognition that the liver is a lymphoid organ with unique immunological properties. These properties ensure efficient innate defence against intestinal microbes and toxins, confer a particular capacity for induction of tolerance, and provide for apoptotic disposal of redundant lymphocytes. Pathological responses within the liver are elicited when: (i) hepatotropic viruses (hepatitis virus B and C) escape immune elimination and reside in hepatocytes; (ii) the liver becomes the site of autoimmune responses directed against either hepatocytes (autoimmune hepatitis) or biliary ductules (primary biliary cirrhosis); or (iii) the liver in the course of disposal of drugs generates neoantigens that provoke adverse allergic responses. Recent advances in the understanding of the immunopathogenesis of these entities are reviewed. PMID- 11869362 TI - Having it all? Stem cells, haematopoiesis and lymphopoiesis in adult human liver. AB - Because of its location and function, the liver is continuously exposed to large antigenic loads that include pathogens, toxins and tumour cells, as well as harmless dietary and commensal proteins and peptides. Therefore, the liver must be actively immunocompetent and, at the same time, control inappropriate inflammatory responses to dietary and other harmless antigens encountered in the portal circulation. In addition to conventional CD4+ and CD8+ T lymphocytes from the circulation, several specialized lymphoid populations are found in the liver to meet these diverse immunological challenges. These populations display the functional and phenotypic properties of innate cells as well as conventional CD4+ or CD8+ helper and cytotoxic T lymphocytes and B cells. The innate lymphoid cells include gammadeltaTCR+ T cells, B1-B cells and NKT cells as well as large numbers of NK cells. The origin of these cells is unknown, but their murine counterparts have been shown to be capable of differentiation in situ in adult liver. Because haematopoietic stem cells have been found in adult human liver as well as molecular evidence of T-cell maturation, we hypothesize that some resident human hepatic lymphoid cells, particularly those expressing innate phenotypes, also differentiate locally. In particular, it is likely that the adult human liver is an important site of NK cell maturation. In this review, we explore the evidence for an active lymphopoietic role for the normal adult human liver. PMID- 11869363 TI - Recruitment of lymphocytes to the human liver. AB - This review discusses the function and localisation of lymphocytes resident within the human liver, under both physiological and pathological conditions. Through description of the mechanisms that mediate lymphocyte recruitment into tissues, this article explains how hepatic endothelial and epithelial cells regulate the recruitment of specific lymphocyte subpopulations. We illustrate that the expression of adhesion molecules and chemokines is crucial to the control of lymphocyte adhesion. Thus, in the normal liver, adhesion molecules such as vascular adhesion protein-1 (VAP-1), intercellular adhesion molecule-1 (ICAM-1) and intercellular adhesion molecule-2 (ICAM-2), and chemokines such as regulated on activation, normal T cell expressed and secreted (RANTES), monocyte chemoattractant protein-1 (MCP-1), macrophage inflammatory protein-1alpha (MIP 1alpha), interferon gamma inducible protein-10 (IP-10), MIG and interferon inducible T-cell alpha chemoattractant (ITAC) are involved in lymphocyte binding to different endothelial compartments. However, in response to inflammation or injury, additional expression of adhesion molecules such as VCAM-1, p-selectin and e-selectin, as well as higher levels of chemokines, permits the attraction and retention of specific effector populations of lymphocytes. We also discuss the expression and function of a newly defined adhesion protein, (VAP-1), and suggest that the unique functions of this protein may provide therapeutic potential for the treatment of liver disease. PMID- 11869364 TI - Biology and significance of T-cell apoptosis in the liver. AB - The liver has emerged as an organ with distinct immunological properties. In this review, we summarize evidence that shows that the liver can remove apoptotic, or non-apoptotic but activated, CD8+ T cells from the circulation and induce apoptosis in these activated T cells by either active or passive mechanisms. Hepatitis viruses, particularly hepatitis C virus, often establish persistent infection. We review evidence that suggests that these viruses exploit intrahepatic tolerance mechanisms to protect themselves from immune attack. PMID- 11869365 TI - Role of primary intrahepatic T-cell activation in the 'liver tolerance effect'. AB - There is accumulating evidence suggesting that hepatic permeability to both naive and activated T lymphocytes may be unique among the solid organs. The possibility that the liver may act as a site of primary activation for CD8+ T lymphocytes is supported by experimental data and may contribute to some of the unique immunological properties of this organ, particularly its ability to induce antigen-specific tolerance. This review discusses the nature of the liver APC inducing primary T-cell activation within the liver: Kupffer cells, liver dendritic cells, liver sinusoidal endothelial cells and hepatocytes are favourably located to allow physical contact with circulating T lymphocytes. Here, we examine the capability of each cell type to act as APC for naive CD4+ or CD8+ T cells and to induce tolerance. PMID- 11869366 TI - Spontaneous acceptance of liver transplants in rodents: evidence that liver leucocytes induce recipient T-cell death by neglect. AB - In many animal models transplanted livers are not rejected, even when there is a complete MHC mismatch between the donor and recipient and the recipient is not immunosuppressed. This distinguishes liver transplants from other organs, such as kidneys and hearts, which are rapidly rejected in mismatched individuals. Acceptance of transplanted livers in a rat model is not due to the absence of an immune response to the liver and there is a rapid, abortive response that is ultimately exhausted. Donor leucocytes transferred with the liver appear to be responsible for both liver acceptance and the abortive activation of the recipient's T cells. The immune mechanism of liver transplant acceptance appears to be due to 'death by neglect' in which T cells are activated to express IL-2 and IFN-gamma mRNA in the recipient lymphoid tissues, but not at adequate levels within the graft. Subsequently the activated T cells die leading to specific clonal deletion of liver donor-reactive T cells. These findings have important implications for liver transplant patients as immunosuppressive drugs that are given to prevent rejection can also interfere with this form of tolerance. In addition, it might be possible to modify the immunosuppressive drug treatment of transplant patients to promote the process of death by neglect of recipient alloreactive T cells. PMID- 11869367 TI - Control or persistence of hepatitis B virus: the critical role of initial host virus interactions. AB - Following infection with hepatitis B virus (HBV), the period before symptomatic disease is now recognized as a time of dynamic interaction between virus and host. Recent work has shown that this period is the phase of infection during which maximal changes in virus replication and the activation of critical components of the immune system occurs. This suggests that the different outcomes following exposure might be determined during the early phase of infection, before the onset of clinical disease. The hypothesis that small differences in the dynamic relationship between host and virus, such as in the kinetics of HBV replication, may influence the final outcome of infection, will be discussed. PMID- 11869369 TI - Animal models of autoimmune liver disease. AB - Autoimmune liver diseases in humans are characterized by chronic active hepatitis with serum autoantibodies, hypergammaglobulinemia and liver pathology showing necroinflammatory disease and fibrosis. There are an increasing number of autoantigens believed to be associated with various autoimmune liver diseases. This review will briefly outline human autoimmune hepatitis and the immunology of the liver. Various murine models of liver inflammation will be discussed, including transgenic and non-transgenic models, with emphasis on how these models aid in our knowledge of the mechanisms of disease development and chronicity. There are limitations with all of the models, including a preponderance of T-cell focused responses. Murine models do not easily develop fibrosis, a hallmark of autoimmune hepatitis in humans. Different experimental models may not reach the same conclusions with differences between immune responses. However, this multiplicity of responses does not necessarily imply that these models are inappropriate for the study of liver immunology and autoimmune liver diseases, as different autoantigens may induce different liver responses. Knowledge of how the liver differs from other immune organs is essential to further our understanding of liver-specific autoimmunity. The plethora of antigens implicated in autoimmune hepatitis in humans predicts that multiple mechanisms may play a role in precipitating disease in the susceptible individual. PMID- 11869368 TI - Epitopes of the NS3 protein of hepatitis C virus: recognition in HLA-DR4 transgenic mice. AB - Hepatitis C virus (HCV) infects more than 180 million of the world's population and causes a persistent infection that over decades can result in cirrhosis and hepatocellular carcinoma. Treatment is only partially effective and control is likely only with the development of effective vaccines. Currently, only chimpanzees can be infected with HCV and alternative animal and tissue culture models are badly needed. We have used mice transgenic for HLA-DR and human CD4 to analyse the specificity of murine responses to the HCV NS3 antigen in an effort to determine whether the epitopes recognized correspond to those recognized by human T cells. Indeed, determinants mapped in transgenic mice overlap with those in a patient exposed to HCV through infection. This result provides hope that such an animal model may be a useful tool with which to analyse particular aspects of immune responses to HCV in vivo. PMID- 11869370 TI - Absence of microdeletions in the Y chromosome in patients with Prader-Willi syndrome with cryptorchidism. AB - Unilateral or bilateral cryptorchidism is found in 80-100% of male patients with Prader-Willi syndrome (PWS). Recently, the relationship between Yq deletions and cryptorchidism has been assessed. However, the relationship between Yq deletions and PWS patients with cryptorchidism remains unclear. Polymerase chain reaction (PCR) amplification of 51 DNA loci encompassing all of the regions for azoospermia factor (AZF) of the Y chromosome, including the deleted in azoospermia (DAZ) and ribonucleic acid-binding motif (RBM) genes, were examined for microdeletions in 10 PWS males with cryptorchidism and 20 healthy control male subjects. No microdeletions of 51 loci were found in any of the PWS males. The present study therefore suggests that microdeletions in the AZF regions of the Y chromosome do not relate to the occurrence of cryptorchidism in PWS patients. PMID- 11869371 TI - Penile pharmacotesting in diagnosing male erectile dysfunction: evidence for lack of accuracy and specificity. AB - Penile pharmacotesting (PPT) with alprostadil (PGE1) represents the most common diagnostic approach to male erectile dysfunction (ED). A positive response - i.e. normal erectile rigidity of sustained duration - is presumed to exclude venous or arterial pathology with enough accuracy. To test this assumption we compared PPT vs. flowmetric results obtained by colour-duplex Doppler ultrasound (CDDU) in patients (pts) undergoing diagnostic evaluation for ED under conditions of maximal cavernous relaxation. A total of 195 non-consecutive impotent pts were diagnosed after dynamic CDDU as non-vasculogenic (NOR), or having arteriogenic (AR), veno-occlusive (VO) or mixed (MX) ED. Maximal erection obtained after PPT was scored as: type-1 (full tumescence - no sustained rigidity, angle on the abdominal plane >90 degrees), type-2 (sustained partial erection, valid for intromission, angle=90 degrees) and type-3 (sustained full erection, angle <90 degrees). Comparing PPT with flowmetric results, we found that a type-3 response had 20% false negative diagnosis of NOR (17% of AR- and 3% of VO- and MX-ED, respectively), while a type-2 response had 63% false negative diagnosis (20% of AR, 37% of VO- and 6% MX-ED, respectively). Type-1 response was associated with the presence of VO dysfunction in 99% of cases. These data suggest that a positive response to PPT (type-2 and type-3) assessed by the visual rating of erection is associated with both arterial (up to 20%) and/or VO (up to 43%) ED, as detected by CDDU. We conclude that PPT alone is a misleading diagnostic test to exclude vascular ED and that dynamic CDDU should be offered to pts investigated for male ED. PMID- 11869372 TI - Are germ cell factors essential in the testicular enlargement after neonatal hypothyroidism recovery? A study using W/Wv mutant mice model. AB - We examined the issue of whether germ cell factors are required for testicular enlargement that occurs after recovery from neonatal hypothyroidism. Experiments were performed using W/Wv mutant mice (lacking germ cells) and normal mice (ICR). The pups in experimental group (neonatal hypothyroid) received 6 propyl 2-thio uracil (PTU) treatment, administered by adding 0.1% (w/v) to the water provided to the mother from day 1 of birth through day 25 postpartum, while the pups of control group received drinking water only. Mice were sacrificed at the age of day 25, 50 and 90, in the case of ICR mice, or at day 25 and 90 in the case of W/Wv mutant mice. In both groups, early hypothyroidism caused a partial recoverable decrease in body growth and testicular development. Both ICR and W/Wv mutant mice, those recovered from neonatal hypothyroidism showed an increase in testis weights, the number of Sertoli cells, and the diameter of the semniferous tubules. This study demonstrates that neonatal hypothyroidism led recovery caused testicular enlargement not only in ICR mice but also in germ cell depleted W/Wv mutant mice. Hence these findings deny direct involvement of the germ cell factors in the process of testicular enlargement in recovered mice even in vivo, and reaffirm the notion that thyroid hormone directly regulates the dynamics of Sertoli cell maturation. PMID- 11869373 TI - Oxytocin promotes spermiation and sperm transfer in the mouse. AB - Spermatogenesis is a complex process during which developing germ cells move from the base of the seminiferous tubule towards the lumen where they are shed. Studies in the rat suggest that seminiferous tubule contraction, induced by exogenous oxytocin, promotes spermiation. This study examines the role of testicular oxytocin in development of the testes, spermatogenesis and spermiation in the mouse. Groups of wild-type (WT) mice, oxytocin knockout mice (OTKO) deficient in testicular oxytocin and mice containing an oxytocin transgene (bOT4.2) that over express testicular oxytocin were killed between days 5 and 45 post partum. The testes and epididymides were removed weighed and prepared either for histological and morphometric study by light microscopy, for sperm counts (epididymis), or extracted for determination of oxytocin content (testis - day 45 only). Testicular oxytocin concentrations were significantly greater (p < 0.05) in bOT4.2 mice than in WT or OTKO mice. No differences in testicular and epididymal weight, or in diameter and area of seminiferous tubules between the mice genotypes were found at any given time. Germ cell development was similar in all genotypes and was comparable with previous studies. The timing of spermiation between the groups was significantly different (p < 0.001) with bOT4.2 < WT < OTKO and the appearance of epididymal sperm was significantly different (p < 0.05) with bOT4.2 < WT < OTKO. There were significant correlations between the percentage of tubules containing residual bodies and epididymal sperm count (p < 0.05) and between the percentage of animals containing residual bodies and the percentage of animals containing epididymal sperm (p < 0.01). These data suggest that in the mouse oxytocin, whilst not involved in germ cell development, is important in the process of spermiation and sperm transfer in the mouse. PMID- 11869374 TI - The clinical studies of sildenafil for the ageing male. AB - Erectile dysfunction (ED) represents an important quality-of-life issue for many ageing men. Low serum testosterone level and other factors may be involved. Sildenafil is effective and well tolerated in patients with ED of various aetiologies, showing an efficacy of about 75%. However, few efficacy and adverse effect studies have focused specially on ageing men. In 150 patients below 65 years and 44 patients over 65 years, sildenafil was used to treat ED. Efficacy in the younger group (89.1%) was greater than in the older group (65.7%; p < 0.01). Mean serum luteinizing hormone (LH) and follicle-stimulating hormone concentrations (11.0 and 18.9 mIU/mL, respectively) in the older group were higher than in younger group (5.2 and 8.7 mIU/mL, respectively; p < 0.01). Serum testosterone and prolactin (PRL) were similar between groups. Older patients showed higher prevalence of diabetes mellitus, hypertension, and benign prostatic hyperplasia. Only diabetes appeared to decrease efficacy of sildenafil in older patients (p=0.019). A high prevalence of diabetes might be one of the many causes of lower efficacy rate of sildenafil in older men, although efficacy in patients even without diabetes in older men was relatively low. Sildenafil can be used safely and it is still effective for ageing male, because nearly two-thirds of our older subjects had a good response to the drug and no adverse effect was specific to older patients. PMID- 11869375 TI - Quantification of telomerase activity, porphobilinogen deaminase and human telomerase reverse transcriptase mRNA in testicular tissue - new parameters for a molecular diagnostic classification of spermatogenesis disorders. AB - The aim of the study was to evaluate the quantitative detection of human telomerase reverse transcriptase (hTERT) mRNA and telomerase activity as new molecular diagnostic parameters for a subclassification of spermatogenesis disorders. Telomerase activity was detected by a quantitative telomerase PCR ELISA, and hTERT mRNA expression was quantified by fluorescence real-time RT-PCR in a LightCycler in cryopreserved testicular tissue specimens. This was paralleled by a histological workup. The discriminant analysis showed that detection of normalized hTERT expression was able to correctly classify 89.0% of the investigated tissue specimens into the subgroups of full spermatogenesis, maturation arrest or Sertoli-cell-only syndrome. In contrast, discriminant analysis revealed an only 58% accuracy of telomerase activity for the investigated tissue specimens. This study shows that the quantification of hTERT expression in testicular tissue by real-time fluorescence RT-PCR is well suited for correctly classifying spermatogenesis disorders and proved to be markedly superior to the determination of telomerase activity. PMID- 11869376 TI - Accuracy of the normal sperm morphology value by Sperm Quality Analyzer IIC: comparison with the strict criteria. AB - The aim of the present study is to investigate the accuracy of the normal sperm morphology value by Sperm Quality Analyzer IIC (SQA IIC), which was developed to provide a rapid and low-cost quantitative evaluation of semen quality. Normal sperm morphology was assessed using SQA IIC in comparison with that by the strict criteria in 62 semen samples. Normal sperm morphology value by SQA IIC was based on the studies of three traditional sperm parameters from over 4000 fresh, untreated semen samples, while the strict criteria was based on the method by Kruger et al. The mean +/- SD of percent normal morphology by SQA IIC and the strict criteria were 37.6 +/- 10.9% (range 15-52) and 19.9 +/- 8.2 (range 1-34), respectively. There was a significant correlation of the sperm morphology assessment between the two methods (r=0.454, p < 0.001). Using the cut-off value of >30% normal morphology by SQA IIC, the positive predictive value and the negative predictive value of the 'normal' strict criteria were 79.6% (39/49) and 46.2% (6/13), respectively. These results indicate that SQA IIC might be used as an initial screening test for the evaluation of sperm morphology. However, sperm morphological assessment by the strict criteria should be performed in order to make decisions in planning strategies for the treatment of infertile couples. PMID- 11869377 TI - Comparison of the Sperm Quality Analyzer IIC variables with the computer-aided sperm analysis estimates. AB - Sperm Quality Analyzer (SQA) IIC, an upgrade version, is an inexpensive device and provides a quantitative estimation of sperm motility, whereas the use of computer-aided sperm analysis (CASA) provides high precision and provision of quantitative data on sperm kinetics. The aim of the present study was to evaluate if the SQA IIC variables correlated with the CASA estimates. Semen quality analysis of 71 fresh semen samples was performed using SQA IIC and CASA. Total sperm concentration, percentage of progressively motile sperm, percentage of normal morphology, motile sperm concentration, sperm motility index (SMI) and functional sperm count (FSC) determinations were performed using SQA IIC. Sperm concentration, sperm motility, and sperm motion variables including amplitude of lateral head displacement (ALH), beat cross frequency (BCF), curvilinear velocity (VCL), straight line velocity (VSL), average path velocity (VAP), linearity (LIN=VSL/VCL), and straightness (STR=VSL/VAP) were evaluated simultaneously on the same semen samples using CASA. The sperm characteristics were compared between SQA IIC and CASA. There were significant correlations of sperm concentration (r=0.634, p < 0.0001), sperm motility (r=0.697, p < 0.0001), and motile sperm concentration (r=0.757, p < 0.0001) between the two devices. Both SMI and FSC significantly correlated with eight CASA estimates, including sperm concentration, sperm motility, motile sperm concentration, ALH, VCL, VSL, VAP, and Rapid. SQA IIC is simple and easy to use. Moreover, the SQA IIC variables well correlated with the CASA estimates. As a screening test for semen quality, SQA IIC is considered as useful in the management of male infertility. PMID- 11869378 TI - A novel homozygous disruptive mutation in the SRD5A2-gene in a partially virilized patient with 5alpha-reductase deficiency. AB - Steroid 5alpha-reductase deficiency is a rare autosomal recessive disorder caused by mutations in the SRD5A2-gene, resulting in diminished dihydrotestosterone (DHT) formation and, hence, in a severe virilization deficit of the external genitalia in patients with 46,XY karyotype. The phenotype of affected individuals is variable and has been reported to range from completely female over genital ambiguity to normal male, depending on the type of mutation and its effect on enzyme activity. Here we report an adolescent 46,XY patient with predominantly female appearance, who had been gonadectomized in early infancy. Genital status revealed a urogenital sinus equivalent to Prader stage III. Molecular genetic analysis demonstrated a homozygous point mutation in exon 2 of the SRD5A2-gene, leading to a premature termination in codon position 111 of the 5alpha-reductase 2 enzyme, and not allowing formation of a functional 5alpha-reductase type 2 enzyme. This case demonstrates that even despite a complete loss of function of 5alpha-reductase type 2, marked virilization is possible, most likely the result of a testosterone (T) effect during foetal life. PMID- 11869379 TI - A preliminary report on the implication of RT-PCR detection of DAZ, RBMY1, USP9Y and Protamine-2 mRNA in testicular biopsy samples from azoospermic men. AB - In this study, reverse transcription-polymerase chain reaction (RT-PCR) was optimized to analyse the presence of DAZ, RBMY1, USP9Y, protamine-2, SRY and actin messenger RNA (mRNA) in testicular cells of men suffering from idiopathic azoospermia. All samples (n=28), including five controls, showed normal expression of actin, SRY and USP9Y. Sperm was not recovered from eight patients after testicular biopsy. Of these, four patients showed altered mRNA levels for the fertility genes, DAZ, RBMY1 and protamine-2. One patient, who was previously shown to be azoospermia factor region (AZF)b deleted, lacked RBM mRNA and presented with reduced amplification of protamine-2 mRNA. This correlated with previous studies, which proposed that RBM expression is exclusive to AZFb and that the lack of testicular RBMY1 mRNA results in suppressed spermatogenesis. Two patients were each lacking DAZ mRNA but did show expression of RBMY1 mRNA at a reduced level, suggesting that there might be residual spermatogenesis in the absence of DAZ expression. Protamine-2 mRNA was detected in one patient and was absent in the second patient. Finally, one patient lacked DAZ, RBMY1 and protamine-2 mRNA. The 19 remaining azoospermic patients presented with normal expression patterns for each of the fertility genes studied. This study demonstrates that the expression of spermatogenesis-specific genes varies in azoospermia. The study of the expression of such genes in a larger number of patients might be useful in characterizing and identifying subpopulations of azoospermic men. PMID- 11869380 TI - Subjective judgements of quality of life: a comparison study between people with intellectual disability and those without disability. AB - The present study investigated the relationship between objective and subjective measures of quality of life (QoL), and in particular, health status and primary care services. Thirty-one people with intellectual disability (ID) were interviewed using a QoL questionnaire. Thirty-one matched controls from the general population filled in a parallel questionnaire. The results in both groups support the findings of many researchers which indicate that satisfaction with aspects of life is generally high (i.e. approximate 75% of maximum possible satisfaction). The present study also confirms previous findings which indicate that satisfaction with aspects of life does not generally correlate with objective circumstances in either group. The importance that people attach to aspects of their lives did not generally correlate with their objective circumstances or their satisfaction with life. However, people with ID attached greater importance to all aspects of their lives than those without disability. This may be linked to their aspirations, preferences and opportunities for choice, and therefore, these factors may be a more meaningful way of considering their QoL. The present findings call into question the use of satisfaction as a general measure of QoL, and also indicate that the use of importance as a mediating variable in understanding satisfaction may not be a solution. The participants with ID had poorer health than the controls, and were significantly less satisfied with their health. Contrary to other findings, the present participants with ID reported that they had received more health checks than the controls over the previous year. Half of these checks had been carried out by 'special' services rather than primary care services. PMID- 11869381 TI - Contraception of women with intellectual disability: prevalence and determinants. AB - The contraception of women with intellectual disability (ID) is a major concern for caregivers. However, the prevalence of contraception and the frequency of use of different methods (e.g. sterilization) remain generally unknown. Moreover, indications specific to women with ID are controversial. The present authors conducted a population-based study among 97% of the women with ID aged between 18 and 46 years attending government-funded facilities in Brussels and the nearby province of Walloon Brabant in Belgium. Out of 397 subjects, 40.8% did not use any contraceptive method, 22.2% were sterilized, 18.4% used an oral contraceptive agent, 17.6% used depotmedroxyprogesterone acetate and 1% used an intrauterine device. These figures differ widely from those of the general Belgian population. Binary logistic regression for 'contraceptive utilization' showed the strong influence of institutional factors such as sleeping environment (i.e. institutional or parental), sexual relationship policy and contraception policy. Having or having had a boyfriend is also correlated with a stronger probability of contraceptive use. Other factors have a smaller influence (e.g. a milder level of ID). Very few factors, none of which are medical, are correlated with an increased use of a specific method. The present results are discussed in the light of the general medical application of contraception and the commonly assumed specific indications for women with ID. PMID- 11869382 TI - Getting personal: an exploratory study of intimate and personal care provision for people with profound and multiple intellectual disabilities. AB - Intimate and personal care is a major area of support and provision for people with intellectual disability (ID), particularly those with profound and multiple ID. However, its management and practice has largely been neglected outside the use of individual guidelines and same-gender intimate care policies, with little research evidence or theoretical literature to inform the planning, conduct and organization of associated care tasks. The present paper reports on the methodology and findings of an exploratory study designed to map the key management and practice issues, and suggest ways forward for the providers of services for people with ID in relation to the quality and outcomes of intimate and personal care. PMID- 11869383 TI - A survey of the prevalence of stereotypy, self-injury and aggression in children and young adults with Cri du Chat syndrome. AB - The aim of the present study was to determine the prevalence and frequency of stereotypy, self-injurious behaviour (SIB), and aggression in children and adults with Cri du Chat syndrome (CCS), and to investigate the relationship between SIB, aggressive behaviour and stereotypy in these individuals. Sixty-six families of children and adults diagnosed with CCS completed the Behaviour Problems Inventory. Additional information relating to gender, chronological age, type of school/post-school occupation and medication was also included in the survey. Stereotyped behaviour was reported for 82% of subjects, more than half the sample displaying it on a daily basis. The occurrence percentage of 15 topographies of SIB suggested that head banging, hitting the head against body parts, self-biting and rumination are the most frequently occurring behaviours in CCS. Aggressive behaviour was reported for 88%, with a statistically significant negative correlation between age and the number of aggressive behaviours reported. The present findings suggest that specific types of stereotypy and SIB are observed frequently in CCS. PMID- 11869384 TI - Prevalence of Helicobacter pylori in people with intellectual disability in a residential care centre in Israel. AB - In 1982, Helicobacter pylori (formerly Campylobacter pylori) was identified as a pathogenic factor in peptic ulcer disease by researchers from Australia. Because only a small number of studies of H. pylori infection have been conducted in people with intellectual disability (ID), and none of these were done in Israel, the present authors decided to conduct a pilot study on its prevalence in this population. The Israeli Division for Mental Retardation provides services to over 6000 people in 54 residential care centres (or institutions), and one centre in the south of the country with kibbutz-style living arrangements was selected for this pilot study. The study was performed as part of the yearly routine medical examination of all residents, and blood specimens were drawn for IgG antibodies to H. pylori (ELISA). Out of the 47 individuals screened, 75% (n=36) were seropositive. In addition to the reported high rates of H. pylori infection in residents with ID living in large facilities, the present pilot study suggests that people with ID living in smaller, kibbutz-style arrangements are also at high risk. PMID- 11869385 TI - Behavioural knowledge, causal beliefs and self-efficacy as predictors of special educators' emotional reactions to challenging behaviours. AB - Theoretical models and emerging empirical data suggest that the emotional reactions of staff to challenging behaviours may affect their responses to challenging behaviours and their psychological well-being. However, there have been few studies focusing on factors related to staff emotional reactions. Seventy staff working in educational environments with children with intellectual disability and/or autism completed a self-report questionnaire that measured demographic factors, behavioural causal beliefs, behavioural knowledge, perceived self-efficacy, and emotional reactions to challenging behaviours. Regression analyses revealed that behavioural causal beliefs were a positive predictor, and self-efficacy and behavioural knowledge were negative predictors of negative emotional reactions to challenging behaviours. Staff with formal qualifications also reported more negative emotional reactions. No other demographic factors emerged as significant predictors. The results suggest that behavioural causal beliefs, low self-efficacy and low behavioural knowledge may make staff vulnerable to experiencing negative emotional reactions to challenging behaviours. Researchers and clinicians need to address these issues in staff who work with people with challenging behaviours. PMID- 11869387 TI - Paid employment of mothers and fathers of an adult child with multiple disabilities. AB - Paid employment is increasingly undertaken by mothers as their children age, with the majority of women being in employment by the time their offspring are adult. Opportunities to engage in employment appear to be reduced for mothers of children with disabilities; however, little is known about the employment of mothers or fathers of adults with disabilities. Data were collected regarding the employment decisions of parents of a young adult with multiple disabilities and contrasted with those of parents whose children were all developing normally. Twenty-five mothers and 12 fathers of a young adult with multiple disabilities were interviewed, as were 25 comparison mothers and 19 comparison fathers. Data collected included hours of work, reasons for employment status, attitudes towards work and child care, and psychological well-being. Clear differences were found between the two groups. Mothers and fathers of a child with multiple disabilities showed different engagement patterns with the paid workforce from comparison parents. Hours of work for fathers of a young adult with multiple disabilities showed a bi-modal distribution, with some fathers working fewer hours than usual and others working very long hours. For mothers in both groups, the number of hours in paid employment was negatively associated with reports of psychological problems. Increased attention needs to be given to the employment opportunities of parents of children with disabilities since employment appears to play a protective role for mothers, in particular. Services provided to adults with disabilities will need to change if parents are to have the same life chances as parents without adult offspring with a disability. PMID- 11869386 TI - Measuring staff support in services for people with intellectual disability: the Staff Support and Satisfaction Questionnaire, Version 2. AB - Social support is an important determinant of well-being, including the stress experienced within the work setting. The present paper reports on the development of the Staff Support and Satisfaction Questionnaire (3SQ), from a previously published measure: the Staff Support Questionnaire. The 3SQ was piloted with 21 health professionals and examined for evidence of test-retest reliability with 24 staff. Data on internal reliability were collected on three occasions from a total sample of 177 staff. The validity of the 3SQ was examined in four studies with a total of 238 staff by testing it against validated measures of psychological well-being. The data showed that the total scale had a high level of test--retest reliability (rs=0.82, P < 0.001) and consistently high internal reliability. Three out of the four validity studies showed statistically significant inverse relationships between the total scale and measures of psychological well-being. The weakest link was the 'Supportive People' subscale. The reliability and validity studies suggest that the psychometric properties of the 3SQ are generally robust, except for the 'Supportive People' subscale, which should be interpreted with caution. PMID- 11869388 TI - Healthcare costs of intellectual disability in the Netherlands: a cost-of-illness perspective. AB - Healthcare costs are continuously increasing, and impose a strong responsibility on governments for an adequate allocation of resources among healthcare provisions and patients. The aims of the present study were to describe the healthcare costs of intellectual disability (ID) and other mental disorders in the context of the total costs of all other diseases, and to determinate the future need of healthcare resources, especially for ID and mental disorders. The present authors performed a top-down cost-of-illness study comprising all healthcare costs of the Netherlands in 1994. Data on healthcare use were obtained for all 22 healthcare sectors, and used to ascribe costs to disease groups, age and sex. Costs of mental disorders are by far the largest in the Dutch healthcare system. Some 25.8% of total disease-specific costs could be ascribed to mental disorders: psychiatric conditions, 10.6%; ID, 9.0%; and dementia, 6.2%. There are large differences between age and sex groups. The costs of ID and schizophrenia are higher among men, and the costs of dementia and depression are higher among women. The age pattern shows two peaks: the first occurs at 25-35 years of age (ID and psychiatric conditions); and the second at 75-85 years of age (dementia). Time trends between 1988 and 1994 show an average annual growth rate of 5.2% for total healthcare costs: psychiatric conditions, 4.8%; ID, 5.4%; and dementia, 9.4%. Demographic projections suggest a less-than-average cost increase for ID and psychiatric disorders (with annual growth rates of 0.2% and 0.4%, respectively) compared to the costs of dementia and total healthcare (with annual growth rates of 1.6% and 0.9%, respectively). Intellectual disability and mental disorders represent a large part of healthcare use in the Netherlands. The costs will inevitably increase because of the ageing of the population and increasing life expectancy among people with disabilities. Non-specific cost containment measures may endanger the quality of care for vulnerable people at younger and older ages. PMID- 11869389 TI - Children with intellectual disability in rural South Africa: prevalence and associated disability. AB - The objective of the present study was to determine the prevalence of intellectual disability (ID) and its associated disabilities in rural South African children aged 2-9 years. It was undertaken in eight villages in the district of Bushbuckridge, Northern Province, South Africa. A two-phase design was utilized. The first phase involved screening children on a house-to-house basis by interviewing mothers or caregivers using an internationally validated questionnaire for detecting childhood disability in developing countries. The second phase consisted of a paediatric/neurodevelopmental assessment of the children who screened positive. A total of 6692 children were screened; 722 (10.8%) had a paediatric evaluation and 238 children were diagnosed with ID, giving a minimum observed prevalence of 35.6 per 1000 children in this population. The prevalence of severe and mild ID was 0.64 per 1000 and 29.1 per 1000 children, respectively. The male:female ratio of children with ID was 3:2. In the affected children, a congenital aetiology for the ID was determined in 49 subjects (20.6%), an acquired aetiology in 15 (6.3%) and the aetiology was undetermined in 174 children (73.1%). Epilepsy (15.5%) and cerebral palsy (8.4%) were the commonest associated disabilities. The present study represents the first data on the prevalence of ID and associated disabilities in rural South African children. The prevalence of ID was comparable with results from a study performed in one other African country (Zambia) as well as those from other developing countries. The data provide an initial factual insight into ID and its associated disabilities for healthcare, social service and educational policy planners. This study provides a basis for the initiation and development of appropriate and integrated services for the best possible care of individuals affected with these disabilities, and for their possible prevention. PMID- 11869390 TI - The ethics of behaviour modification: a comment on ethical issues in the implementation of a DRI programme for the treatment of social isolation and ritualistic behaviour in a learning disabled individual. Reinders J. S. The good life for citizens with intellectual disability. Journal of Intellectual Disability Research 46, 1-5. PMID- 11869391 TI - Emerging viral infections in Australia. AB - Emerging viruses include known viruses that have increased in incidence or geographic range (such as enteroviruses and Japanese encephalitis virus), new viruses associated with known diseases (Australian bat lyssavirus) and new viruses associated with previously unrecognized diseases (Hendra and Nipah viruses). Some may have a predilection for children (Japanese encephalitis, influenza viruses and enterovirus 71) and vigilance is essential to ensure early recognition of these agents. PMID- 11869392 TI - Indigenous neonatal outcomes: What do we expect? PMID- 11869393 TI - Paediatric telephone advice: a major gap in quality service delivery. PMID- 11869394 TI - Australian Paediatric Surveillance Unit : progress report. AB - The Australian Paediatric Surveillance Unit (APSU), through active surveillance, collects information on the epidemiology of rare or uncommon childhood conditions. This research resource allows paediatricians to collaborate at State, national and international levels. The APSU sends a monthly report card to all (currently 971) paediatricians in Australia, who in turn indicate whether or not they have seen a patient in the last month with any of the conditions listed (98% response rate in 1999). Study investigators, notified by the APSU of positive case reports, obtain demographic and clinical data on the patient from the reporting doctor by postal questionnaire (90% response rate in 1999). Since 1993, the APSU has monitored 27 conditions, including conditions that are vaccine preventable, otherwise infectious, genetic, congenital and non-communicable. Information collected is disseminated to paediatricians and other health professionals via an annual report, newsletters and publications and is made available to the International Network of Paediatric Surveillance Units. Information provided by the APSU has raised awareness among paediatricians of rare and uncommon childhood conditions, and has been used by health authorities for planning of prevention and intervention strategies and allocation of health resources. PMID- 11869395 TI - Is being Aboriginal or Torres Strait Islander a risk factor for poor neonatal outcome in a tertiary referral unit in north Queensland? AB - OBJECTIVE: To compare perinatal outcomes for all births, and the morbidity and mortality patterns of babies admitted to neonatal intensive care, for non Indigenous, Aboriginal and Torres Strait Islander (TSI) people in a major remote urban centre. METHODS: A prospective cohort study of all births and consecutive admissions to the Neonatal Intensive Care Unit/Special Baby Care Unit (NICU/SCBU) at Kirwan Hospital for Women (KWH), Townsville (a tertiary referral hospital) was conducted from January 1998 to June 1999. RESULTS: There were 2575 births, with 2392 (93%) booked into KWH for delivery. The Aboriginal group differed from the non-Indigenous group only in the prevalence of low-birthweight births. The TSI group had significantly higher perinatal mortality rates, preterm and low birthweight births than the non-Indigenous group. Seven hundred and fifty-eight babies were admitted to NICU/SCBU: 586 (77%) non-Indigenous, 129 (17%) Aboriginal, 43 (6%) TSI. There were significantly more preterm, lower birthweight and longer stay length Aboriginal and TSI babies. Being Indigenous was not associated with neonatal death. Gestation of less than 28 weeks, congenital anomalies, and high-grade cerebral haemorrhage, but not ethnicity, were independent risk factors for neonatal death. Maternal risk factors, including poor antenatal care attendance, were more prevalent amongst Indigenous women. CONCLUSION: Neonatal outcomes for Aboriginal infants were better than expected from national and State reports. Outcomes for Torres Strait Islander infants were worse than expected. Ethnicity was not a risk factor for neonatal death. These findings suggest that outcomes may be further improved by programmes to increase access for Indigenous women to antenatal care services. PMID- 11869396 TI - Professional telephone advice to parents with sick children: time for quality control! AB - OBJECTIVE: To assess the quality of professional telephone advice given to parents with sick children. METHODS: All hospitals with an emergency department and a paediatric ward and a designated child health telephone advice line in the greater Brisbane region were invited to participate in the study. Case scenarios involving a febrile baby, a 14-month-old with gastroenteritis and an 18-month-old with a head injury were used three times with each institution. Each of the cases should have elicited a response indicating the need for urgent medical attention. A research assistant presented the symptoms in accordance with the questions of the telephone advice-giver. Aspects of the call were recorded, including time between call made and access to advice-giver, profession of advice-giver, identifying details sought by advice-giver, questions asked about the case presented, length of call, and advice-given. RESULTS: Of the 10 hospitals asked, six agreed to participate. Included in the study were two paediatric hospitals, two general public hospitals, one private hospital, and a Statewide-designated child health telephone advice line. Calls were generally attended to promptly. Only 37 (68.5%) of the advisers recognized the urgency of the case scenarios, with the febrile infant having the least likelihood of eliciting the correct advice (39%). Doctors gave more reliable advice than nurses in the fever scenario, but not in the other cases. CONCLUSIONS: Telephone advice to parents with sick children is easily accessible, but is often dangerously inappropriate. There is a need for tighter quality control and/or for a centralised telephone advice line to prevent inappropriate advice being given to parents. PMID- 11869397 TI - Children with disruptive behaviours I: service utilization. AB - OBJECTIVE: To describe the demographic characteristics and patterns of service utilization in the Hunter region (NSW, Australia) of families caring for a child manifesting disruptive behaviour. METHODOLOGY: Families were eligible to participate in the survey if they had at least one child known to have one of the DSM-IV disruptive behaviour disorders, autistic spectrum disorders, behaviour problems associated with rarer forms of brain disease, brain injury or mild intellectual disability, or identified by school personnel as having significant behaviour problems. Families were recruited to the survey via schools, early education centres and clinical services. Parents completed a mail survey about demographic details and service utilization. RESULTS: In total, 1412 families responded to the survey, a participation rate of approximately 65%. The demographic characteristics of respondent families were similar to those of the regional population. A high proportion (85%) of children from respondent families were receiving treatment. Nearly half those children attending clinical services were attending two or more services. Approximately one-third each of the sample was attending public clinics, private clinics or a combination of both. Children were more likely to be treated by a paediatrician (64%) than a child psychiatrist (39%) or a psychologist (23%). Compared with a normative Australian sample, parents of children manifesting disruptive behaviour reported a high level of family stress. CONCLUSIONS: Families of children manifesting disruptive behaviour are a highly stressed group who often access several clinical services. Paediatricians make a significant contribution to the management of children with disruptive behaviours and need to be considered in service planning. PMID- 11869398 TI - Children with disruptive behaviours II: clinical and community service needs. AB - OBJECTIVE: To assist in health service planning by determining the perceived clinical and community service needs of families resident in the Hunter region who care for a child manifesting disruptive behaviour. METHODOLOGY: Families were eligible to participate in the survey if they had at least one child known to have one of the DSM-IV disruptive behaviour disorders, autistic spectrum disorders, behaviour problems associated with rarer forms of brain disease, brain injury or mild intellectual disability or were identified by school personnel as having significant behaviour problems. Families were recruited to the survey via schools, early education centres and clinical services. Parents completed a questionnaire, mailed to them by educational and/or clinical services. Parents were asked to prioritize options for improving or expanding clinical services and for reducing their treatment costs. RESULTS: A total of 1412 families responded to the survey. The highest-ranked clinical service options involved the expansion of mainstream community treatment services for children with disruptive behaviour problems and their families, with a particular emphasis on counselling services. Respite care and in-patient services were given relatively low priority. Subgroup analyses showed that disadvantaged and stressed families gave higher rankings to out of home options, such as respite care, in-patient care and subsidised holiday camps, than the aggregate sample. CONCLUSIONS: Enhancement of community based counselling services would meet the needs of the greatest number of participating families. Resource-intense residential services are required by a small but important group of families who experience disadvantage and high levels of stress. PMID- 11869399 TI - Evaluation of a community intervention programme for preschool behaviour problems. AB - OBJECTIVES: To evaluate an early intervention project focused on improving child prereading skills and parent behaviour-management skills, for 4-year-old children in the western suburbs of Melbourne (Victoria, Australia). METHODOLOGY: Following screening of a large sample of preschoolers, four groups were constructed: (i) children receiving a phonological skills-based prereading package in their normal preschool programme; (ii) children whose parents attended six sessions of child behaviour management skill training; (iii) children who received both interventions; and (iv) a control group. Pre-, post-, 1 and 2 year follow-up measures were taken to assess effects on child behavioural and learning adjustment. RESULTS AND CONCLUSIONS: This community based intervention resulted in positive effects, despite being of low intensity, in achieving significantly less hyperactive/distractable behaviour in children from the combined intervention group at 2 years postintervention. Anxiety problems were also reduced at the post-treatment phase. The prereading package did not demonstrate any effects on reading skills and was deemed to be insufficiently intensive to affect a population of disadvantaged preschoolers. This universal-type of intervention was well accepted by the community, but there is need for further effort to increase recruitment of families of at-risk children into such programmes. PMID- 11869400 TI - Shopping trolley-related injuries to children in New Zealand, 1988-97. AB - OBJECTIVE: To describe the epidemiology of shopping trolley related injuries (fatalities and hospitalizations) to children in New Zealand prior to the introduction of a voluntary standard for shopping trolleys. METHODOLOGY: To identify cases, a key word search was conducted of national mortality and hospitalization databases for the years 1988-97. Cases were limited to children under 15 years of age. RESULTS: For the 10 year period investigated, 282 hospitalizations and no fatalities were identified. A significant increasing trend for hospitalizations was detected (chi2 = 17.6, 1 d.f.; P < 0.001). Ninety two per cent of children hospitalized were aged under 5 years and two-thirds were aged 2 years or younger. Ninety per cent of injuries resulted from falls from trolleys, 84% of injuries were to the head or face and 22% were rated serious (AIS-3) on the Abbreviated Injury Scale. CONCLUSIONS: The incidence of injuries associated with shopping trolleys increased between 1988 and 1997. Following the introduction of a voluntary standard for shopping trolleys in 1999, which included specifications for child harnesses, trends in injury should be monitored. PMID- 11869401 TI - Birthweight and stature, body mass index and fat distribution of 14-year-old Polish adolescents. AB - OBJECTIVES: The relationships between intra-uterine growth retardation and stature, relative weight and fat distribution at adolescence have not been comprehensively established. The aim of this report is to assess the effect of low birthweight on stature, relative weight and fat distribution in 14-year-old boys and girls from Wroclaw, Poland. METHODOLOGY: Cross-sectional measurements of 1197 boys and 819 post-menarcheal girls aged 13.50-14.49 years were performed during medical examinations in 1997. Stature, body mass index (BMI; kg/m2), waist to-hip ratio (WHR) and waist-to-thigh ratio (WTR) were used in the present study. A cut-off value of the 10th percentile of birthweight for particular gestational weeks was used in order to define subjects born small for gestational age (SGA) or appropriate for gestational age (AGA). Two-way ANOVA was used to evaluate the effect of birthweight on anthropometric variables of 14-year-old adolescents, allowing for socioeconomic status (determined by the level of the mother's education). RESULTS: Birthweight affected stature in boys and girls (P < 0.001), BMI in boys (P < 0.05) and WHR and WTR in girls (P < 0.001 and P < 0.05, respectively). At the age of 14 years, both SGA boys and girls were shorter than their AGA peers. The SGA boys had lower BMI, whereas SGA girls accumulated more centralized fat compared with their AGA counterparts. CONCLUSION: Fetal growth retardation has a long-lasting adverse effect on later physical growth. Polish SGA children do not catch up with their peers in terms of stature by adolescence. Moreover, central fat distribution, as observed among SGA girls, constitutes a significant risk for several adult degenerative diseases. PMID- 11869402 TI - Growth and adrenal suppression in asthmatic children on moderate to high doses of fluticasone propionate. AB - OBJECTIVE: Growth and adrenal suppression have been reported in asthmatic children using high-dose inhaled fluticasone propionate (FP). Inhaled FP, given at moderate doses (250-750 microg/day), has not been documented to be associated with growth or adrenal suppression in asthmatic children until recently. We report three cases illustrating these side effects. METHODS: Growth and adrenal suppression, after the introduction of inhaled FP, were observed in three prepubertal young asthmatic children referred to our asthma clinic and growth clinic. Growth centile and velocity were assessed by longitudinal stadiometry height measurements. Early morning plasma cortisol levels, and glucagon stimulation tests were used to assess the pituitary adrenal axis. RESULTS: Severe growth and adrenal suppression were noted in three children while they were on moderate doses of inhaled FP. Improvements in growth and adrenal function were observed following cessation or dose reduction of inhaled FP. CONCLUSIONS: Unexpected growth and adrenal suppression may occur in young asthmatic children using moderate doses of inhaled FP. PMID- 11869403 TI - Pseudomonas fluorescens pseudobacteraemia: a cautionary lesson. AB - OBJECTIVES: To describe an outbreak of pseudobacteraemia caused by Pseudomonas fluorescens in a paediatric population. To document and highlight the effect this outbreak had on clinical management and the steps taken to determine the source. METHODS: A clinical and microbiological investigation was carried out into a cluster of 38 pseudobacteraemias caused by Pseudomonas fluorescens in paediatric patients over a 10 month period. RESULTS: The source of the outbreak of pseudobacteraemia was found to be contaminated lithium heparin tubes, which were being filled prior to the filling of the blood culture bottle. Cultures of the same tubes yielded Pseudomonas fluorescens. As a result of the initial positive blood cultures, clinical management was altered in 18 cases. A staff education programme was instituted and eventually resulted in a cessation of the pseudobacteraemia. CONCLUSIONS: Pseudobacteraemias are a major cause of potentially inappropriate therapy in febrile children. Attention to detail in the collection of blood cultures can help reduce this outcome. Staff involved in the collection of blood cultures need to be aware of this potential source of contamination. PMID- 11869404 TI - Sleep deprivation, disorganization and fragmentation during opiate withdrawal in newborns. AB - OBJECTIVE: To determine specific sleep characteristics in neonatal opiate withdrawal, referred to as the Neonatal Abstinence Syndrome (NAS), by measuring sleep efficiency, deprivation, disorganization and fragmentation in three groups: (i) healthy term neonates; (ii) opiate-exposed neonates who were treated for opiate withdrawal; and (iii) a group of opiate-exposed neonates who did not require treatment. METHODS: A cohort study recording sleep patterns of neonates at 2-10 days of age (after 36 or more weeks of gestation) was carried out. Twenty one neonates were exposed to opiates during pregnancy and 15 neonates were healthy controls. Sleep characteristics were predefined, and treated newborns were divided into early and stabilized treatment groups. Polygraphic recordings of sleep, movement and breathing were made continuously after a daytime feed. RESULTS: Sleep deprivation, disorganization and fragmentation were found in newborns with NAS and were associated with the severity of the withdrawal. Neonates treated for NAS displayed increased wakefulness during early treatment (deprivation), but were similar to controls once stabilized. Both treated and non treated groups had reduced amounts of quiet sleep (deprivation). Treated newborns showed an increase in indeterminate sleep (disorganization) and arousals-to wakefulness (fragmentation). CONCLUSION: This study determined the exact nature and degree of sleep disturbances in newborns during acute opiate withdrawal. The findings contribute to a further understanding of the physiology underlying neonatal opiate withdrawal and suggest that some changes in sleep are due to opiate withdrawal but others may reflect opiate dependency in utero. PMID- 11869405 TI - Eating behaviour of children 1.5-3.5 years born preterm: parents' perceptions. AB - OBJECTIVE: To investigate parents' perceptions of the eating behaviour of toddlers born preterm. METHODS: A mailed, self-administered questionnaire was sent to parents of infants born between July 1994 and July 1996 with gestational ages less than 32 weeks who had been admitted to the Neonatal Unit at Flinders Medical Centre. RESULTS: Responses were received for 95 subjects (response rate 70%) with a mean +/- SD gestational age of 29.2 +/- 2.1 weeks, birthweight of 1243 +/- 296 g and median corrected age of 134 weeks (range 84-185 weeks) at the time of study. Seventy-eight per cent of parents were concerned about the quality of food intake and 45% wished to change their child's eating behaviour. Although only 20% of parents identified their toddlers as being fussy eaters, food refusal was reported by 58% of respondents, and 51% and 69% used food rewards or coaxing, respectively, to encourage intake. Parents of toddlers who had received home gavage feeding (n = 5) or had a respiratory or neurological disability (n = 16), reported poorer feeding indicators. CONCLUSIONS: Programmes providing information on normal toddler eating behaviour and strategies to promote positive feeding practices may reassure parents and promote development of food preferences and eating behaviours consistent with long-term healthy eating habits. Special emphasis should be placed on children with a disability or children who have received home gavage feeding. PMID- 11869406 TI - Telephone reminders improve adolescent clinic attendance: a randomized controlled trial. AB - OBJECTIVES: To measure the effect of telephone reminders on adolescent clinic attendance. METHODS: Clinic bookings of adolescents were randomly assigned to either a telephone reminder one day prior to their appointment, or a routine booking (no reminder). The setting was four general adolescent health clinics within a tertiary public adolescent health care service at the Centre for Adolescent Health. The main outcome measures were clinic non-attendance, reason for non-attendance, and satisfaction with the booking system. RESULTS: One hundred and seventy one adolescent appointments were studied. Of these, 51.5% were female, and 25% of bookings were for new, rather than review appointments. One hundred and one adolescents were randomly allocated to the reminder group, of whom 87% were contacted. The use of reminders (intention to treat analysis) significantly reduced the non-attendance rate from 20% to 8% (odds ratio 0.35; P = 0.03). Non-attendance was three times more likely for a new appointment than for review appointments. 'Forgetting' was the most common explanation given by patients (35%) who did not attend. Seventy-nine per cent of parents reported telephone reminders were helpful at prompting attendance. CONCLUSION: Telephone reminders greatly improved attendance at these adolescent clinics. The background non-attendance rate and the proportion of high-risk patients for non-attendance (new appointments in this setting) will determine whether reminders are more efficiently targeted at specific bookings than used routinely. PMID- 11869407 TI - Use of hypoallergenic formula in the prevention of atopic disease among Asian children. AB - OBJECTIVE: To determine the effect of a partially hydrolysed formula on genetically predisposed children, with respect to the development of atopic clinical manifestations and in vitro testing of serum IgE levels (total and milk specific). METHODS: One hundred and ten infants were randomly assigned to receive either partially hydrolysed formula or standard infant formula, and were prospectively monitored from birth for clinical atopic symptoms and serum IgE levels. RESULTS: Eczema occurred less frequently in infants receiving partially hydrolysed formula. This was significant (P < 0.05) at 3, 6, 9, 12, 18 and 24 months. However, the significance decreased with time, although it almost reached statistical significance at 30 months by the Kaplan-Meier survival function (log rank statistic, 3.46; P = 0.063). Although wheezing occurred less frequently in infants receiving partially hydrolysed formula, compared to those receiving standard infant formula, the difference did not reach statistical significance (P > 0.05). CONCLUSIONS: Exclusive feeding of hypoallergenic milk formula in the first 4 months of life has a protective effect in terms of the development of atopic dermatitis in the first 2 years of life, compared to feeding with cow's milk formula. PMID- 11869409 TI - Recurrent parotitis. PMID- 11869408 TI - An unexpected cause of jaundice. PMID- 11869410 TI - Drug-induced extrapyramidal reactions. AB - A child with psychotic symptoms and attention-deficit hyperactivity disorder who developed extrapyramidal symptoms while on a combination of risperidone, methylphenidate, sertraline, tropisetron and ketorolac is described herein. The extrapyramidal symptoms resolved with the administration of benztropine, an anticholinergic drug. Successful treatment of his psychosis was achieved by decreasing the dose of risperidone, followed by slow upward titration. PMID- 11869412 TI - Diaphragmatic paralysis in extremely low birthweight neonates: Is waiting for spontaneous recovery justified? AB - Mortality and morbidity associated with surgical management of patent ductus arteriosus (PDA) in neonates has been reported to vary from 0% to 44%. Complications like pneumothorax, pleural effusion, recurrent nerve and phrenic nerve injury are associated with surgical closure of PDA. An extremely low birthweight neonate with diaphragmatic paralysis following phrenic nerve injury during surgical closure of PDA is reported. Delay in diaphragmatic plication for over two weeks while waiting for spontaneous recovery was associated with significant morbidity including chronic lung disease. The controversies associated with timing of diaphragmatic plication in high-risk neonates are discussed. PMID- 11869411 TI - Cardiac involvement in infantile Sandhoff disease. AB - An 18-month-old boy with enzyme assay-confirmed infantile Sandhoff disease (MIM 268800) is reported. Besides the classical neurological features, this patient exhibited severe mitral regurgitation secondary to mitral valve prolapse and mild aortic regurgitation from aortic valve prolapse. He also had asymmetric hypertrophy of the interventricular septum without left ventricular outflow tract obstruction. PMID- 11869413 TI - Audit and cost of treatment modalities provided for respiratory syncytial virus bronchiolitis in a Malaysian teaching hospital. PMID- 11869414 TI - Congenital cytomegalovirus infection associated with finger anomaly. PMID- 11869415 TI - Asthma is not prevalent in Aboriginal and Torres Strait Islander children: a myth. PMID- 11869416 TI - Neonatologists and echocardiography. PMID- 11869417 TI - Echocardiographic misdiagnosis and ultrasound skills. PMID- 11869418 TI - Diagnostic accuracy of paediatric echocardiograms. PMID- 11869421 TI - Obstructive sleep apnea and the prefrontal cortex: towards a comprehensive model linking nocturnal upper airway obstruction to daytime cognitive and behavioral deficits. AB - Obstructive sleep apnea (OSA) is accompanied by significant daytime cognitive and behavioral deficits that extend beyond the effects of sleepiness. This article outlines a causal model by which to understand these psychological effects among OSA patients. The model proposes that sleep disruption and blood gas abnormalities prevent sleep-related restorative processes, and further induce chemical and structural central nervous system cellular injury. This, in turn, leads to dysfunction of prefrontal regions of the brain cortex (PFC), manifested behaviorally in what neuropsychologists have termed 'executive dysfunction'. Executive dysfunction is proposed to markedly affect the functional application of cognitive abilities, resulting in maladaptive daytime behaviors. The proposed model (1) accounts for the specific psychological phenotype associated with OSA, (2) accommodates developmental components in this phenotype, (3) bridges between physical and psychological phenomena, (4) suggests mechanisms by which the nocturnal disorder might have effects on daytime functioning, (5) is empirically testable, (6) generates unique research hypotheses, and (7) has practical implications. The model is intended to act as a catalyst for future research and as a preliminary guide for clinicians. PMID- 11869422 TI - Long-term vs. short-term processes regulating REM sleep. AB - In cats, rats, and mice, the amount of rapid eye movement sleep (REMS) lost during a sleep deprivation (SD) predicts the subsequent REMS rebound during recovery sleep. This suggests that REMS is homeostatically regulated and that a need or pressure for REMS accumulates in its absence, i.e. during both wakefulness and non-rapid eye movement sleep (NREMS). Conversely, it has been proposed that REMS pressure accumulates exclusively during NREMS [Benington and Heller, Am. J. Physiol. 266 (1994) R1992; Prog. Neurobiol. 44 (1994b) 433]. This hypothesis is based on the analysis of the duration of successive NREMS and REMS episodes and of electroencephalogram (EEG) events preceding REMS. Pre-REMS events (PREs) do not always result in sustained REMS and can thus be regarded as REMS attempts that increase as NREMS progresses. It is assumed that two processes regulating REMS can resolve the apparent contradiction between these two concepts: a 'long-term' process that homeostatically regulates the daily REMS amount and a 'short-term' process that regulates the NREM--REMS cycle. These issues were addressed in two SD experiments in rats. The two SDs varied in length (12 and 24 h) and resulted in very similar compensatory changes in NREMS but evoked very different changes for all REMS parameters studied. The large REMS increase observed after 24-h SD was accompanied by a reduction in unsuccessful PREs and an increase in sustained REMS episodes, together resulting in a threefold increase in the success-rate to enter REMS. Changes in success-rate matched those of a theoretically derived long-term REMS pressure. The SD induced changes in sleep architecture could be reproduced by assuming that the increased long-term REMS pressure interacts with the short-term process by increasing the probability to enter and remain in REMS. PMID- 11869423 TI - Debating how REM sleep is regulated (and by what). PMID- 11869425 TI - Spindle frequencies in sleep EEG show U-shape within first four NREM sleep episodes. AB - It has been shown in previous studies on sleep electroencephalogram (EEG) that spindles are slower in the beginning of the night fastening towards the end of the night. Corresponding findings have been obtained by spectral analysis. The present study was based on our preliminary observation that slower spindles are found in the middle of the non-rapid eye movement (NREM) sleep episodes as compared with the beginning or the end of the episodes. Eight healthy female and six male subjects were studied. Sleep spindles were visually selected and spindle frequencies calculated for 11 analysis points in each NREM sleep episode. The median spindle frequencies formed a clear U-shape within NREM sleep episodes with an initial decrease and final increase. The decrease was statistically significant within the first four NREM sleep episodes. It is possible that the spindle frequency pattern could be used to reveal variations in sleep depth within sleep stage 2. In animal studies the spindle frequency has been found to be associated to the duration of the hyperpolarization-rebound sequences of the thalamocortical cells. If it is assumed that the same cellular mechanisms are responsible for spindle frequencies in humans then the study of variations in spindle frequency could be used to examine the NREM sleep process in humans. PMID- 11869426 TI - Spatial organization of EEG activity from alertness to sleep stage 2 in old and younger subjects. AB - In order to elucidate brain mechanisms that contribute to the increased tendency for vigilance dysregulation in the elderly, we examined the spatial organization of brain electric activity [electroencephalogram (EEG)] during decreasing vigilance from alertness to onset of sleep stage 2, comparing 7 old and 10 younger, healthy subjects (60-79 and 18-41 years old, respectively). Two features were analyzed: (1) change of location of the brain electric source gravity centers of the EEG frequency bands, and (2) magnitude of fluctuation of these locations over time. Multichannel EEG was analyzed into source gravity center localizations for seven EEG frequency bands, using fast Fourier transform (FFT) Dipole Approximation (first principal component-single source modeling in the frequency domain). Multivariate analysis of covariance (MANCOVA) showed: source localizations were more anterior in old than younger subjects for beta-1 and more superior for all three beta bands; from alertness to sleep, delta and theta EEG sources (inhibitory activity) changed to more posterior and superior areas, and alpha-1 and -2 (routine activity) and beta-1 and -2 sources (excitatory activity) towards anterior and superior areas. Fluctuations of the source locations of delta and beta-2 were larger on the superior--inferior axis, and of beta-2 smaller on the left-right axis in the old than younger subjects. The results suggest functional specifications (inhibitory, routine, excitatory) of cortical positron emission tomography (PET) changes reported in sleep. In sum, aging exhibits changes in spatial organization of EEG-generating neuronal assemblies; during the transition wakefulness-to-sleep, aging affects the spatial-temporal dynamics of this organization. The latter is suggested to contribute to the increased risk for consciousness disturbances in the elderly. PMID- 11869427 TI - Sleep- and circadian-dependent modulation of REM density. AB - Rapid eye movement (REM) density, a measure of the frequency of rapid eye movements during REM sleep, is known to increase over the course of the sleep episode. However, the circadian modulation of REM density has not been thoroughly evaluated. Data from a forced desynchrony protocol, in which 20 consecutive sleep opportunities were systematically scheduled over the entire circadian cycle, were analysed. The REM density was evaluated from polysomnographically recorded REM sleep episodes, and analyzed as a function of time in the sleep opportunity and as a function of phase in the circadian cycle. The REM density showed a robust increase over the course of the sleep episode. This sleep-dependent increase was observed regardless of circadian phase, because data analyzed from different thirds of the circadian cycle exhibited a similar pattern. The REM density did not show a significant circadian-dependent modulation for data from the entire sleep opportunity. However, analysis of circadian modulation from separate thirds of the sleep opportunity revealed a significant circadian modulation in the last third of the sleep episode. Maximum REM densities were observed when the last third of the sleep episode coincided with the wake-maintenance zone, i.e.;8-10 h before the crest of the circadian rhythm of REM sleep propensity. These results confirm the dominant sleep-dependent modulation of REM density, and indicate that the density of REMs is greatest when sleep pressure is low, such as in the latter part of the sleep episode, at which time the circadian modulation of REM density is also appreciable. PMID- 11869428 TI - Sleep propensity varies with behaviour and the situation in which it is measured: the concept of somnificity. AB - It is self-evident that we fall asleep more readily when lying down than when standing up. Nevertheless, the influence of this and more subtle changes in posture, activity and the situation in which sleep propensity is measured have been largely ignored. The term somnificity is introduced here, defined as the general characteristic of a posture, activity and situation that reflects its capacity to facilitate sleep-onset in a majority of subjects. The relative somnificities of different activities and situations in the Epworth Sleepiness Scale (ESS) were investigated in 23 groups, involving 2802 subjects from seven different countries. The means of the different ESS item-scores were ranked from highest to lowest in each group. There was a high concordance (Kendall's C=0.84, P < 0.0001) among these ranks for all groups, whether of normal subjects or patients with sleep disorders, regardless of age, sex, or average sleepiness in daily life assessed by total Epworth scores. The ESS item-ranks formed an ordinal scale of somnificities with five different levels. Analysis of raw ESS item scores for all 987 individual Australian subjects showed the same pattern of somnificities with six different levels, but with a lower concordance (C=0.39, P < 0.0001). This was probably because of subject x situation-specific interactions that were averaged within groups. A conceptual model of sleepiness is outlined that includes interactions between separate sleep and wake drives as a possible way of including behavioural and situational influences on sleep propensity. PMID- 11869429 TI - A prospective study of fatal occupational accidents -- relationship to sleeping difficulties and occupational factors. AB - Very little is known about the association between sleep and (fatal) occupational accidents. This study investigated this relationship using register data of self rated sleep difficulties, together with occupational and demographic characteristics. The variables were related to subsequent occupational fatal accidents. A national sample of 47,860 individuals was selected at regular intervals over a period of 20 years, and interviewed over the phone on issues related to work and health. The responses were linked to the cause of death register (suicides excluded) and the data set was subjected to a (multivariate) Cox regression survival analysis. One hundred and sixty six fatal occupational accidents occurred, and the significant predictors were: male vs. female: relative risk (RR)=2.30 with a 95% confidence interval (CI) of 1.56-3.38; difficulties in sleeping (past 2 weeks): RR=1.89 with CI=1.22-2.94; and non-day work: RR=1.63 with CI=1.09-2.45. No significant effect was seen for age, socio economic group, hectic work, overtime (>50 h per week), or physically strenuous work. It was concluded that self-reported disturbed sleep is a predictor of accidental death at work, in addition to non-day work and male gender. PMID- 11869430 TI - Time-courses in renin and blood pressure during sleep in humans. AB - We previously described a strong concordance between nocturnal oscillations in plasma renin activity (PRA) and the rapid eye movement (REM) and non-REM (NREM) sleep cycles, but the mechanisms inducing PRA oscillations remain to be identified. This study was designed to examine whether they are linked to sleep stage-related changes in arterial blood pressure (ABP). Analysis of sleep electroencephalographic (EEG) activity in the delta frequency band, intra arterial pressure, and PRA measured every 10 min was performed in eight healthy subjects. Simultaneously, the ratio of low frequency power to low frequency power + high frequency power [LF/(LF + HF)] was calculated using spectral analysis of R -R intervals. The cascade of physiological events that led to increased renin release during NREM sleep could be characterized. First, the LF/(LF + HF) ratio significantly (P < 10(-4) decreased, indicating a reduction in sympathetic tone, concomitantly to a significant (P < 10(- 3) decrease in mean arterial pressure (MAP). Delta wave activity increased (P < 10(-4) 10-20 min later and was associated with a lag of 0-10 min with a significant rise in PRA (P < 10(-4) . Rapid eye movement sleep was characterized by a significant increase (P < 10(-4) in the LF/(LF + HF) ratio and a decrease (P < 10(-4) in delta wave activity and PRA, whereas MAP levels were highly variable. Overnight cross-correlation analysis revealed that MAP was inversely correlated with delta wave activity and with PRA (P < 0.01 in all subjects but one). These results suggest that pressure dependent mechanisms elicit the nocturnal PRA oscillations rather than common central processes controlling both the generation of slow waves and the release of renin from the kidney. PMID- 11869431 TI - Short-term homeostasis of REM sleep assessed in an intermittent REM sleep deprivation protocol in the rat. AB - An intermittent rapid eye movement (REM) sleep deprivation protocol was applied to determine whether an increase in REM sleep propensity occurs throughout an interval without REM sleep comparable with the spontaneous sleep cycle of the rat. Seven chronically implanted rats under a 12 : 12 light-dark schedule were subjected to an intermittent REM sleep deprivation protocol that started at hour 6 after lights-on and lasted for 3 h. It consisted of six instances of a 10-min REM sleep permission window alternating with a 20-min REM sleep deprivation window. REM sleep increased throughout the protocol, so that total REM sleep in the two REM sleep permission windows of the third hour became comparable with that expected in the corresponding baseline hour. Attempted REM sleep transitions were already increased in the second deprivation window. Attempted transitions to REM sleep were more frequent in the second than in the first half of any 20-min deprivation window. From one deprivation window to the next, transitions to REM sleep changed in correspondence to the amount of REM sleep in the permission window in-between. Our results suggest that: (i) REM sleep pressure increases throughout a time segment similar in duration to a spontaneous interval without REM sleep; (ii) it diminishes during REM sleep occurrence; and (iii) that drop is proportional to the intervening amount of REM sleep. These results are consistent with a homeostatic REM sleep regulatory mechanism that operates in the time scale of spontaneous sleep cycle. PMID- 11869432 TI - CSF hypocretin-1 (orexin-A) concentrations in narcolepsy with and without cataplexy and idiopathic hypersomnia. AB - We measured cerebrospinal fluid (CSF) hypocretin-1 levels in 11 patients with narcolepsy-cataplexy, five with narcolepsy without cataplexy and 12 with idiopathic hypersomnia (IHS). All patients were Japanese. As reported in Caucasian patients, undetectable or very low hypocretin-1 levels were observed in most (9 out of 11) Japanese narcolepsy--cataplexy patients. Our hypocretin deficient narcoleptics included three prepubertal cases within few months after the disease onset. All nine hypocretin-deficient patients were human leuckocyte antigen (HLA) DR2 positive, while two who had normal CSF hypocretin-1 levels were HLA DR2 negative. In contrast, none of the narcolepsy without cataplexy and IHS subjects had undetectable low levels. Low CSF hypocretin-1 is therefore very specific for HLA DR2 positive narcolepsy-cataplexy, and the deficiency is likely to be established at the early stage of the disease. PMID- 11869434 TI - 'Arts and humanities': a new section in Medical Education. PMID- 11869435 TI - Communication skills training: pragmatism versus proof. PMID- 11869436 TI - Challenges in educational research. PMID- 11869437 TI - Do what I say, not what I do. PMID- 11869438 TI - Implementing a communication skills programme in medical school: needs assessment and programme change. AB - INTRODUCTION: Communication skills training (CST) in medicine, once considered a minor subject, is now ranked a core clinical skill. To assess the state of formal and informal CST at Dalhousie Medical School a needs assessment was undertaken in 1997 with the goal of using these findings to plan and implement a new communication skills curriculum. OBJECTIVES: This article briefly describes the relevant findings of the needs assessment, the subsequent development of an integrated cross curriculum CST programme, and early programme evaluation results. METHOD: Surveys were completed by undergraduates at the end of pre clinical (n=65), and clinical phases (n=82), residents (n=54), and faculty (n=117). Results revealed learners' and faculty's appreciation of the importance of CST, learners' assessment of training weaknesses in the delivery of CST, learners' weakness in higher order patient--doctor communication skills, and faculty weakness in assessing learners' communication skills competency. The results also indicated that CST was generally not being addressed either formally or informally in clinical medical education. RESULTS: The paper describes and discusses the subsequent implementation (beginning in 1998) of CST into the medical school curriculum. There is a description of programme development and evaluation at the pre-clinical, clerkship and postgraduate levels, a description and discussion of faculty development, and discussion of the importance of financial and administrative support for the programme. CONCLUSION: Programme evaluation results at all levels are positive. PMID- 11869439 TI - The effectiveness of a computer-assisted instruction programme on communication skills of medical specialists in oncology. AB - BACKGROUND: Although doctor--patient communication is important in health care, medical specialists are generally not well trained in communication skills. Conventional training programmes are generally time consuming and hard to fit into busy working schedules of medical specialists. A computer-assisted instruction (CAI) programme was developed -- 'Interact-Cancer' -- which is a time efficient learning method and easily accessible at the workplace. OBJECTIVE: To investigate the effect of the CAI training, 'Interact-Cancer', on the communication behaviour of medical specialists, and on satisfaction of patients about their physician interaction. DESIGN: Consultations of medical specialists with cancer outpatients were videotaped at 4 specific stages, 2 before and 2 after Interact-Cancer, with intervals of 4 weeks. PATIENTS/PARTICIPANTS: Participants were 21 medical specialists, mainly internists, working in 7 hospitals, and 385 cancer outpatients. METHODS: Communication behaviour was assessed on 23 observation categories derived from the course content. Frequencies were rated as well as judgements about the quality of the performance of each target skill. Satisfaction was measured by the Medical Interview Satisfaction Scale. Data were analyzed by means of multilevel statistical methods. RESULTS: The behavioural assessment showed course effects on ratings of the physicians' quality of performance. No course effects were found on the frequencies of physicians' behaviours and on the patient satisfaction ratings. CONCLUSIONS: CAI is a promising method to supply medical specialists with postgraduate training of communication skills. The application of judgement ratings of communication behaviour proved to be valuable to evaluate course effects in real-life patient encounters. PMID- 11869440 TI - Medical students as standardized patients to assess interviewing skills for pain evaluation. AB - PURPOSE: The use of medical students as standardized patients in a performance assessment of pain evaluation was studied. METHODS: Fifty-two pairs of second year medical students participated. One student portrayed a patient presenting with cancer pain and was interviewed by the other medical student. The student patient then rated the interview using a checklist of pain assessment and general interviewing skills. The interviews were audiotaped and also rated independently. RESULTS: Based on student-patient ratings, 36 (69%) students demonstrated 9 or more of the 11 pain-specific checklist items, compared to 34 (65%) students according to the trained rater. Highly specific pain-related items had higher agreement than broader interviewing skill items. There would be differences in the summary assessments of students depending on which rating data were used. DISCUSSION: Medical students represent a readily accessible resource as patients for clinical simulations. Students tended to overestimate the performance of fellow students, but acting as a standardized patient had educational value, and can be used to extend simulated patient encounters within the curriculum. Further investigation is needed to improve the reliability of the feedback provided by student-patients. PMID- 11869441 TI - The development of a scale to measure medical students' attitudes towards communication skills learning: the Communication Skills Attitude Scale (CSAS). AB - INTRODUCTION: There is little research identifying medical students' attitudes towards communication skills learning. This pilot study outlines the development of a new scale to measure attitudes towards communication skills learning. METHODS: First- and second-year medical students (n = 490) completed the 26-item Communication Skills Attitude Scale (CSAS) and 39 students completed the CSAS on a second occasion. Factor analysis was conducted to determine the factors underpinning the scale. The internal consistency of the subscales was determined using alpha coefficients. The test-retest reliability of the individual scale items were determined using weighted kappa coefficients and the test-retest reliability of the subscales were established using intraclass correlation coefficients. RESULTS: Maximum likelihood extraction with direct oblimin rotation resulted in a 2-factor scale with 13 items on each subscale. Factor I represented positive attitudes towards communication skills learning and factor II represented negative attitudes. Subscale I had an internal consistency of alpha=0.873 and an intraclass correlation of 0.646 (P < 0.001). Subscale II had an internal consistency of alpha=0.805 and an intraclass correlation of 0.771 (P < 0.001). The majority of items on the positive (n=9, 69.2%) and the negative attitude subscales (n=8, 61.5%) possessed moderate test-retest reliability. DISCUSSION: The development of a new and reliable scale to identify medical students' attitudes towards communication skills learning will enable researchers to explore the relationships between medical students' attitudes and their demographic and education-related characteristics. Further work is needed to validate this scale among a broader population of medical students. PMID- 11869442 TI - Predicting communication skills with a paper-and-pencil test. AB - AIM: This study was conducted to investigate the value of a written knowledge test of communication skills for predicting scores on a performance test of communication skills. METHOD: A paper-and-pencil test of knowledge about communication skills and a performance test of communication skills, consisting of four stations with standardised patients, were administered to students of two classes of the medical schools of Maastricht and Leiden, the Netherlands. The results on these tests were compared. RESULTS: From the results of both instruments, the classes of the participating students could be recognised equally well: 60% correct qualifications of the classes by the knowledge test and 64% by the multiple station examination. Between the two tests an overall, disattenuated correlation of 0.60 was found (N=133, P < 0.01), suggesting moderate predictive value of the knowledge test for the performance test of communication skills. The correlation is stronger for students from Maastricht medical school than for their colleagues in Leiden. Correlation between the knowledge of communication skills test and other available test results of the participating Maastricht students is close to zero, suggesting that the test measures a distinct quality of students' competence. DISCUSSION: The paper-and pencil test of knowledge of communication skills has predictive value for the performance of these skills, but this value seems to be less pronounced than similar findings for clinical procedural skills. The stronger relationship between 'knowing how' and 'showing' in the Maastricht student group might be indicative of an effect of the training format. PMID- 11869443 TI - The influence of admissions variables on first year medical school performance: a study from Newcastle University, Australia. AB - AIMS: This study examined the relationship between the performance of first year medical students at the University of Newcastle, Australia, and admission variables: previous educational experience, and entry classification (standard -- academic or composite, Aboriginal and Torres Strait Islander, or overseas), age and gender. METHODS: Admission and demographic information was obtained for students who entered first year medicine at Newcastle between the years 1994 and 1997 inclusive. Academic performance was measured according to results of first assessment ('satisfactory' vs. 'not satisfactory') and the final assessment of the first year ('satisfactory' vs. 'not satisfactory'). Logistic regression was used to examine the relationship between predictor variables and outcomes. RESULTS: Assessment and admissions information was obtained for 278 students, 98% of all students who entered the medical course between 1994 and 1997. Regression analysis of first assessment indicated that Aboriginal and Torres Strait Islander and overseas students were significantly more likely to be 'not satisfactory' than all other students (RR=3.1,95% CI: 1.4. - 6 7 and RR=1.5, 95% CI: 1.2-1.8, respectively). Analysis of final assessment indicated these two student groups were also significantly more likely to be 'not satisfactory' than all other students (RR=4.5, 95% CI: 1.4-13.5 and RR=3.5, 95% CI: 1.2-10.8, respectively). At first assessment, students entering via the standard academic pathway and older students were less likely to be 'not satisfactory' (RR=0.6, 95% CI: 0.5-0.7 and RR=0.8, 95% CI: 0.7-0.9, respectively). However both these differences were not evident at final assessment. There were no significant relationships between performance in first year and the remaining variables. CONCLUSIONS: Aboriginal and Torres Strait Islander, and overseas medical students had academic difficulties in the first year of the course, suggesting the need for extra course support. The result may reflect the educational and other obstacles these students must overcome in order to enter and progress through their medical degree. More research is warranted to explore the extent to which these differences persist throughout the medical degree. PMID- 11869444 TI - Adapting the Key Features Examination for a clinical clerkship. AB - PURPOSE: A written test of clinical decision-making, the Key Features Examination, was developed for use in clerkship. METHODS: Following the guidelines provided by the Medical Council of Canada, a Key Features Examination was developed and implemented in an internal medicine clinical clerkship, during the 1998/99 clerkship year. The reliability and concurrent validity of the exam were assessed. RESULTS: A 2 hour examination, containing 15 key feature problems, was administered to 101 students during 6 consecutive internal medicine clerkship rotations. The reliability of the exam, calculated from Cronbach's alpha, was 0.49. The exam had modest correlation with other measures of knowledge and clinical performance. CONCLUSION: The Key Feature Examination is a feasible and reliable evaluation tool that may be implemented as a component of student assessment during a clinical clerkship. PMID- 11869445 TI - Collusion detection in multiple choice examinations. AB - OBJECTIVE: To develop and test a novel method for collusion detection in multiple choice examinations. SUBJECTS AND METHODS: Answers from two negatively marked medical prize examinations for two different years were analysed. Both examinations were administered electronically. One examination was formally invigilated, while the second was not; instead, candidates were able to sit the test at any time and at any computer with an internet connection. We examine pairs of students and compare correlations between their answers. Our approach allows us to correct for the difficulty of individual questions and for the estimated ability and the degree of risk aversion of the students. We compare the results of this statistical analysis with other information on the timing of the answers and the physical location of the computer, both of which are available to the web-server. RESULTS: Significant correlation between several candidates who either admitted having cheated or could be linked to other corroborating evidence of collusion was found. CONCLUSION: It is possible to detect collusion in multiple choice examinations in a statistical way by examining the patterns of answers between pairs of candidates. In examinations that are delivered on-line, information is often available on the location of the candidates and the timings of their answers, and can be used as additional corroborative evidence. PMID- 11869446 TI - Students' perceptions of whistle blowing: implications for self-regulation. A questionnaire and focus group survey. AB - AIM: The aims of this study are to gauge and use the views of medical students to elaborate on the concept of whistle blowing and to give an indication of its role in self-regulation. METHODS: An anonymous questionnaire survey was conducted among medical students from a Scottish medical school. Students were asked (i) whether they should, and (ii) whether they would report to faculty academic misconduct by other students. Comparisons were made between all five year groups. Subsequently students in focus groups were asked to give their reasons for whistle-blowing or not doing so. RESULTS: Of the students, 40% felt they should whistle-blow, with 13% saying they would actually do so. The numbers of students who felt they should or would whistle blow were smaller in the later year groups. Focus groups identified positive and negative reasons for whistle blowing or not, and reasons connected to the process itself. Reasons given for not whistle blowing included camaraderie, retaliation by peers, self-preservation and a belief that it is not a student's responsibility to report the misconduct of others. CONCLUSIONS: The results have two important implications. First, medical schools have to decide whether students have a duty to whistle blow and/or whether there is a need to devise clear procedures. Any procedures should take into account the reasons given for not whistle blowing, but should concentrate on positive motivating factors. Secondly the medical profession needs to consider the role of whistle blowing, as the results suggest that whistle blowing should not be the only method of detection of misconduct in an undergraduate setting. PMID- 11869447 TI - Identifying oneself as a teacher: the perceptions of preceptors. AB - INTRODUCTION: Attracting and retaining qualified medical educators will be a continuing challenge as physicians feel more pressure to produce clinical revenue. With this paper we hope to begin to identify how clinical teachers, judged as excellent by their peers, establish their professional identity as physician and teacher. METHOD: This qualitative study examines the views of 10 clinical teachers to identify the characteristics excellent preceptors attribute to their teaching identity. RESULTS: This study revealed four themes that influence faculty teaching identity: underlying humanitarianism, familiarity with adult learning principles, understanding of the benefits and drawbacks of teaching, and the image of self as teacher. CONCLUSION: Using the findings from this study, suggestions are given to faculty developers for activities to increase physician identity as teacher. PMID- 11869448 TI - European convergence towards higher standards in dental education: the DentEd Thematic Network Project. AB - INTRODUCTION: This paper describes a Thematic Network Project (TNP) funded by the European Union's Directorate for Education and Culture. It focused on convergence towards higher standards in dental education in the European Union and included those 'Associate Countries' seeking membership of an expanding EU. The DentEd Project sought to use peer influence in promoting convergence towards higher standards in European dental education and training. There is evidence of serious differences in standards despite the European Union's Dental Directives and the series of guidelines and recommendations of their Advisory Committee on the Training of Dental Practitioners. METHODS: The DentEd TNP sought to improve communication, better understanding of different systems and to pool intellectual resources. A central objective of the network was to promote outcome analysis and the implementation of evidence-based treatments. DentEd sought to identify and disseminate innovations and best practices in European dental schools and stomatological institutions. This was achieved by instigating 30 self-assessments of dental schools throughout Europe. Self-assessment was followed by a visit from a team of international peers. Each assessment comprised 19 sections and was placed on the DentEd web site (www.dented.org) after it was jointly approved by the visitors and host school. The 30 self-assessment reports together with the visitors comments and recommendations were then divided into their component sections and analysed. Responsibility for each section was allocated to one of 19 different international working groups of between 8 and 12 people. Preliminary analyses from these working groups were presented and debated on the DentEd interactive web site. They finalised their findings in the DentEd Report at a Plenary meeting in the Nobel Forum in the Karolinska Institute in Stockholm. The meeting was organised in such a way that each working group finalised their report. This was followed by the rotation of the working groups to listen and respond to the presentation of another group on the basis of a series interactive meetings. At the end of the meeting the original groups reassembled and revised their analysis on the basis of the additional input from the other 200 individuals involved and then finalised their group's report on the web site. These reports are available on the web site and were also published in hard copy.1 The three-year DentEd Project achieved its stated objectives. CONCLUSION: The DentEd Thematic Network Project employed a process of schools visits, plenary meetings and the use of an interactive web site to help reach consensus on innovations, best practices and significant divergence in standards in dental education in Europe. This process has proven an effective method of improving communication and of increasing the understanding of diversity in a wide range of educational methods. It also emphasised the importance of continuing quality improvement and outcome analysis of the clinical competence of graduates from European dental schools. It is important to emphasise that schools visited are likely to reflect those with an open and flexible attitude to educational reform and development and perhaps they were not representative of those schools that would benefit most from a self-assessment followed by peer review. On completion of the DentEd Thematic Network Project (TNP) the European Union's Directorate for Education and Culture approved a new three-year DentEdEvolves TNP and the consortium has recently submitted a major proposal in the EU's eLearning initiative on the basis of the success of this collaboration. This has extended to include 75% of dental schools from North America and plans for extension to South-east Asia will bring a major meeting to Singapore in April 2002. PMID- 11869449 TI - Career destinations and views in 1998 of the doctors who qualified in the United Kingdom in 1993. AB - OBJECTIVE: To report career destinations and views in 1998 of doctors who qualified in the United Kingdom (UK) in 1993. DESIGN: Postal questionnaire survey. SETTING: This study took place in the United Kingdom. SUBJECTS: All doctors who qualified in the UK in 1993. MAIN OUTCOME MEASURES: The percentage of doctors in each branch of medicine five years after qualification, and their views on their training and career opportunities. RESULTS: The NHS and universities in the UK employed 88% of respondents (men 90%, women 86%). UK general practice employed 24% of respondents (men 19%, women 28%). There were significant differences (P < 0.01) between the percentages of men and women working in the surgical specialties (men 28%, women 10%), paediatrics (men 8%, women 15%) and obstetrics and gynaecology (men 5%, women 10%). Respondents not in paid employment comprised 1.4% of men and 6.6% of women. 45% of respondents agreed that their postgraduate training was of a high standard, with 26% disagreeing and 29% unsure. 47% of specialist registrars felt their training was too short and 78% were concerned about the availability of consultant posts on completion. CONCLUSIONS: Although loss of doctors from the British workforce through emigration or unemployment is not increasing, our findings confirm a substantial shift away from careers in general practice. The number of home trained GPs from this generation of doctors will be inadequate to meet service needs. GPs and hospital specialist doctors expressed concerns about quality of training, lack of careers advice, the shortness of specialist registrar training and availability of consultant posts on completion of training. PMID- 11869452 TI - The comparative RNA web (CRW) site: an online database of comparative sequence and structure information for ribosomal, intron, and other RNAs. AB - BACKGROUND: Comparative analysis of RNA sequences is the basis for the detailed and accurate predictions of RNA structure and the determination of phylogenetic relationships for organisms that span the entire phylogenetic tree. Underlying these accomplishments are very large, well-organized, and processed collections of RNA sequences. This data, starting with the sequences organized into a database management system and aligned to reveal their higher-order structure, and patterns of conservation and variation for organisms that span the phylogenetic tree, has been collected and analyzed. This type of information can be fundamental for and have an influence on the study of phylogenetic relationships, RNA structure, and the melding of these two fields. RESULTS: We have prepared a large web site that disseminates our comparative sequence and structure models and data. The four major types of comparative information and systems available for the three ribosomal RNAs (5S, 16S, and 23S rRNA), transfer RNA (tRNA), and two of the catalytic intron RNAs (group I and group II) are: (1) Current Comparative Structure Models; (2) Nucleotide Frequency and Conservation Information; (3) Sequence and Structure Data; and (4) Data Access Systems. CONCLUSIONS: This online RNA sequence and structure information, the result of extensive analysis, interpretation, data collection, and computer program and web development, is accessible at our Comparative RNA Web (CRW) Site http://www.rna.icmb.utexas.edu. In the future, more data and information will be added to these existing categories, new categories will be developed, and additional RNAs will be studied and presented at the CRW Site. PMID- 11869455 TI - Serovar distribution of a DNA sequence involved in the antigenic relationship between Leptospira and equine cornea. AB - BACKGROUND: Horses infected with Leptospira present several clinical disorders, one of them being recurrent uveitis. A common endpoint of equine recurrent uveitis is blindness. Serovar pomona has often been incriminated, although others have also been reported. An antigenic relationship between this bacterium and equine cornea has been described in previous studies. A leptospiral DNA fragment that encodes cross-reacting epitopes was previously cloned and expressed in Escherichia coli. RESULTS: A region of that DNA fragment was subcloned and sequenced. Samples of leptospiral DNA from several sources were analysed by PCR with two primer pairs designed to amplify that region. Reference strains from serovars canicola, icterohaemorrhagiae, pomona, pyrogenes, wolffi, bataviae, sentot, hebdomadis and hardjo rendered products of the expected sizes with both pairs of primers. The specific DNA region was also amplified from isolates from Argentina belonging to serogroups Canicola and Pomona. Both L. biflexa serovar patoc and L. borgpetersenii serovar tarassovi rendered a negative result. CONCLUSIONS: The DNA sequence related to the antigen mimicry with equine cornea was not exclusively found in serovar pomona as it was also detected in several strains of Leptospira belonging to different serovars. The results obtained with L. biflexa serovar patoc strain Patoc I and L. borgpetersenii serovar tarassovi strain Perepelicin suggest that this sequence is not present in these strains, which belong to different genomospecies than those which gave positive results. This is an interesting finding since L. biflexa comprises nonpathogenic strains and serovar tarassovi has not been associated clinically with equine uveitis. PMID- 11869456 TI - Angiotensin converting enzyme inhibitor therapy in children with Alport syndrome: effect on urinary albumin, TGF-beta, and nitrite excretion. AB - BACKGROUND: Angiotensin converting enzyme inhibitors are routinely prescribed to patients with chronic kidney disease because of their known renoprotective effects. We evaluated the effect of short-term therapy with the angiotensin converting enzyme inhibitor, enalapril, in early Alport syndrome, defined as disease duration less than 10 years and a normal glomerular filtration rate. METHODS: 11 children with early Alport syndrome were investigated. Two consecutive early morning urine specimens were collected at the start of the study for measurement of urinary creatinine, total protein, albumin, TGF-beta, and nitrite excretion. Patients were treated with enalapril, congruent with 0.2 mg/kg/day, once a day for 14 days. Two early morning urine specimens were collected on days 13 and 14 of enalapril treatment and two weeks later for measurement of urinary creatinine, total protein, albumin, TGF-beta, and nitrite excretion. RESULTS: Prior to treatment, urinary excretion of transforming growth factor-beta and nitrite, the major metabolite of nitric oxide, was within normal limits in all patients. Administration of enalapril for 2 weeks did not alter urinary albumin, transforming growth factor-beta, or nitrite excretion. CONCLUSION: These findings suggest that early Alport syndrome represents a disease involving exclusively intrinsic glomerular barrier dysfunction. At this stage of the illness, there is no evidence of angiotensin II-mediated proteinuria or increased production of transforming growth factor-beta and, therefore, routine treatment with an angiotensin converting enzyme inhibitor may not be warranted. PMID- 11869454 TI - Enhanced immunogenicity of a functional enzyme by T cell epitope modification. AB - BACKGROUND: T helper epitopes are necessary for the induction of high titers of antigen-specific IgG antibodies. We are interested in the epitope modification of intact proteins as a method to enhance their immunogenicity for the generation of recombinant protein-based vaccines. RESULTS: Hartley strain guinea pig T cell epitopes were mapped for two related bacterial proteases. Two T cell epitopes were found in one of the proteases, while a comparatively reduced immunogenicity protease had no detectable T cell epitopes. A T cell epitope sequence homologous to the immunogenic protease was created in the less immunogenic protease by changing a single amino acid. Proliferative responses to the whole protein parent enzyme were two-fold higher in splenocyte cultures from variant-immunized animals. We found that the single amino acid change in the variant resulted in a protein immunogen that induced higher titers of antigen-specific IgG antibody at low doses and at early time points during the immunization protocol. The serum from parent- and variant-immunized guinea pigs cross-reacted at both the protein and the peptide level. Finally, animals primed to the variant but boosted with the parent enzyme had higher levels of antigen-specific IgG than animals immunized with the parent enzyme alone. CONCLUSIONS: With a single amino acid change we have introduced a T cell epitope into a comparatively low-immunogenic enzyme and have increased its immunogenicity while retaining the enzyme's original proteolytic function. The ability to immunomodulate proteins while leaving their function intact has important implication for the development of recombinant vaccines and protein-based therapeutics. PMID- 11869457 TI - Effects of testicular microlithiasis on Doppler parameters: report of three cases. AB - BACKGROUND: Testicular microlithiasis is a rare, usually asymptomatic, non progressive disease of the testes associated with various genetic anomalies, infertility and testicular tumors. According to our literature search, there is no specific data about Doppler findings in this disease. CASE PRESENTATION: Doppler findings of three cases of testicular microlithiasis during last two years in our institution are presented. CONCLUSIONS: Although our hypothesis was to find increased Doppler parameters due to intratesticular arterial compression, our findings suggest that there are no Doppler findings specific to testicular microlithiasis. PMID- 11869458 TI - Plasma proteins in a standardised skin mini-erosion (II): effects of extraction pressure. AB - BACKGROUND: A standardised suction technique has been used to sample plasma proteins in dermal interstitial fluid (IF) serially for 5 to 6 days from a suction-induced skin mini-erosion. Increased protein concentrations ascribed to inflammation have been shown from day 1 onward. In this study, we assessed the effect of two different extraction pressures on IF sample composition. METHODS: Total protein concentration and the concentrations of insulin, prealbumin, albumin, transferrin, IgG and alpha-2-macroglobulin were assessed daily in healthy volunteers. Samples were extracted at 50 mmHg and 200 mmHg below the atmospheric. RESULTS: At 0 h after forming the erosion, mean total IF protein content (relative to plasma) was lower in the samples extracted at -200 mmHg than at -50 mmHg (26 +/-13% (SD) vs 48 +/-9.8%; p < 0.05). There were no significant differences at 24, 48, 72 or 96 h. Of the individual proteins, expressed as area units (AU) for area under the curve (AUC) from 0-96 h, albumin was lower in IF sampled at -200 mmHg (2.49 +/- 0.68 vs 3.08 +/- 0.36 AU; p < 0.05), as was transferrin (1.91 +/- 0.52 vs 2.40 +/- 0.42 AU; p < 0.05). Extraction volumes were significantly higher at -200 mmHg (AUC diff: 60%; p < 0.05). CONCLUSIONS: Samples of IF extracted at 0 h at -200 mmHg contained lower protein concentrations, indicating an increased water fraction and an intact sieve function of the vascular wall. The difference in protein concentration extracted at higher and lower pressure from 24 h onward was less pronounced. Lower pressure should be used to sample substances of greater molecular size. PMID- 11869459 TI - Stress reactions in police officers after a disaster rescue operation. AB - This study intended to determine the prevalence of posttraumatic stress disorder symptoms and to report subjective well-being, general distress, and social functioning among police officers 18 months after a rescue operation during a fire at a discotheque. Emotional responses, including self-reported reactions, recorded during and after the fire were analysed. Forty-one police officers participated in the study. They completed a questionnaire and a battery of self report measures. Psychological distress was recorded using the PTSS-10, IES-R, and GHQ-28. The three scales had a high internal consistency. The police officers were not unaffected by their experiences, and a few officers were still under stress. One police officer had IES-R intrusion and an avoidance score greater than 20, suggesting a stress reaction of clinical significance. When compared with the PTSS-10 scale, two (5%) officers showed a high level of psychological distress. Measured with the GHQ-28, three officers (7%) still had a high level of psychological distress. On one of the four different subscales most of the officers show various degrees of reduced social functioning. Debriefing was carried out by the police department after the fire, and 75% thought debriefing was positive. Eighteen months after the disaster a few police officers are still under stress. PMID- 11869460 TI - Long-term consequences of the reformation of psychiatric care: a 15-year follow up study. AB - Although there are a great many epidemiological studies of psychiatric diseases and the outcomes of controlled rehabilitation programmes, there is a shortage of studies of routine psychiatric care. This study is a retrospective cohort study of the total number (n=138) of patients receiving inpatient care at a psychiatric rehabilitation clinic in Jonkoping during 1984. The follow-up period is 15 years, 1984-98. The group had a considerable amount of inpatient care, but this decreased dramatically during the time of investigation. Positive indications were that there were no suicides during the last 5 years of the period and that the number of registered crimes showed a marked decrease from 1992. Negative indications were low levels of work and occupation and few social contacts. PMID- 11869461 TI - Are readmission rates influenced by how psychiatric services are organized? AB - A transition is taking place in the organization of psychiatric services. The length of stay in hospitals is reduced and partly replaced by treatment and care in the community. The readmission rate is proposed as one indicator to analyse the effectiveness of this transition. A comprehensive literature review was conducted to ascertain whether readmission rates differ with different service systems. The search yielded 77 articles, of which 28 were selected on the basis of aims and objectives of the articles. The results show that approximately 50% of all patients admitted to psychiatric hospitals are previously admitted patients. Longer length of stay, appropriate discharge planning, and follow-up visits after discharge predicted fewer readmissions, whereas the quantity and quality of community care did not seem to have any impact on readmission rates. On the basis of the analysis of the literature review it is concluded that readmission rates are not a suitable indicator of quality of care in psychiatric hospitals. Readmission rates may, however, be an important tool in the planning of mental health services. PMID- 11869462 TI - Causal and pathoplastic risk factors of depression: findings of the Tampere Depression Project. AB - Factors associated with low self-esteem in non-depressive subjects increase the individual's vulnerability to depression (causal risk factors), and factors correlated to low self-esteem in depressive subjects make the current disorder more severe (pathoplastic risk factors). Using the Rosenberg Self-Esteem Scale and Depression Scale we intended to explore correlates of low self-esteem in non depressive and depressive subjects in a random sample of 1643 individuals attending community health centres in Central Finland. According to our study, self-esteem in non-depressive men was affected mainly by poor socioeconomic situation; in depressive men particularly low self-esteem was associated with negative family factors. Low self-esteem in non-depressive women was correlated to poor socioeconomic situation, poor health, and negative family factors. In depressive women self-esteem was affected by poor physical and poor mental health. Taking into consideration causal and pathoplastic risk factors, general practitioners can improve recognition of depression. By paying attention to pathoplastic factors, it is possible to improve detection of more severe forms of depression. PMID- 11869463 TI - Partial remission in major depression: a two-phase, 12-month prospective study. AB - We conducted an interview-based survey to predict the clinical course of major depressive disorder during a follow-up period of 12 months. Altogether 86 patients were investigated. A SCID I interview for DSM-III-R axis-I diagnosis was conducted at baseline and a SCID II interview for personality disorders at the 6 month follow-up. Beck Depression Inventory scores indicated the level of depression and were compiled at baseline and at 6 and 12 months. A BDI score between 9 and 14 was considered to indicate partial remission, and score of 0-8 indicated remission. At the 6-month assessment 33% of the patients had remission, 20% were in partial remission, and 47% were in the depressive phase. Older age, personality disorder, and alexithymia were associated with poor response at 6 months. At 12 months 37% had remission, 28% were in partial remission, and 35% were still in the depressive phase. Treatment at the early stage should be effective enough to achieve remission. If the response is not satisfactory within 6 months, a renewed search should be conducted for factors hindering recovery. Comorbid personality disorder is the main factor predicting a poor short-term response in major depressive disorder. PMID- 11869464 TI - Characteristics of the social environment of small group homes for individuals with severe mental illness. AB - As part of research into small group homes run by local authorities for those with severe mental illness the characteristics of the social environment was investigated. The ideal and real versions of the Community Oriented Program Environmental Scale (COPES) were administered to residents and staff of small group homes in a county in southern Sweden. Both these groups were also interviewed on their opinions as to the main characteristics that should exist in the social environment in small group homes. High levels of support, order and organization, and program clarity and a low level of anger and aggression distinguished the real social environment according to the residents. Differences between residents' and staff's perceptions were less pronounced than in most other studies, probably owing to a greater level of resident--staff interaction in the home-like milieu. In the interviews both the residents and staff emphasized the importance of social interaction as a major constituent in the social environment of small group homes. The COPES instrument appears to be valid for use in small group homes, although some concern can be raised about two of the subscales, Involvement and Practical Orientation. The social environment, as rated by the residents and staff, was similar to that recommended for psychotic patients and to that proposed as a suitable supportive profile together with low levels of staff control and anger and aggression. PMID- 11869465 TI - Auto-vampirism in schizophrenia. AB - Vampirism and auto-vampirism have been reported for centuries as very rare features. A relationship between vampirism auto-vampirism and mental disorders has been established, especially with regard to schizophrenia. This case study reports auto-vampirism in a 35-year-old schizophrenic woman. The Present State Examination was used to obtain a valid diagnosis. Auto-vampirism proved to be part of a bizarre delusion of a purification process. This patient reported experiences of depersonalization and auditory commanding hallucinations that made her strive for auto-vampirism. Although rare, auto-vampirism can be detected as a symptom in schizophrenia and can be treated. PMID- 11869466 TI - Three cases of angel's trumpet tea-induced psychosis in adolescent substance abusers. AB - An increase of intoxications in persons using alkaloid-containing ornamental plants (mainly angel's trumpet) for their hallucinogenic effects and easy and cheap availability is registered. It is mainly adolescents who are experimenting with these plants. In addition to severe vegetative anticholinergic symptoms, the clinical picture is often dominated by a toxic psychosis with hallucinations, disturbances of orientation, and psychomotoric agitation, aggression, or anxiety. Three cases of severe psychotic pictures with only mild or completely missing vegetative symptoms after ingestion of angel's trumpet tea are reported. Caused by the increasing spreading of angel's trumpet shrubberies in Europe and North America, intoxications with large numbers of fatalities have to be expected in the future. The taxonomical, pharmacological, and clinical aspects of angel's trumpet-induced disorders are discussed. Knowledge of the clinical picture is important for correct diagnosis and treatment. Prevention by mass media should mainly focus on the medical fatalities of abuse and not emphasize the hallucinogenic effects, presumably increasing unwanted interest for these plants among youths. PMID- 11869467 TI - Does extended medication with amphetamine or methylphenidate reduce growth in hyperactive children? AB - The purpose of the present study was to examine growth in children on extended stimulant treatment. Ninety-one hyperactive boys were studied, of whom 68 were treated with amphetamine and 23 with methylphenidate. The age range was 3-10 years. All children were treated with one of these stimulants for a minimum of 1 year. The yearly mean values for height and weight were all between the 25th and 90th percentile of the population norms. During the 1st year of treatment children who used amphetamine showed significantly smaller weight gains than those who used methylphenidate (F = 6.9, df = 64, P < 0.05). Four patients (17%) in the methylphenidate group and 21 (31%) in the amphetamine group had a weight loss during the 1st year, ranging from 0 to 9.5 kg. Within this group of 25 patients there was a significantly higher number of children with a pretreatment weight greater than that of the 50th percentile group (chi-square = 5.59, P < 0.05). All 25 patients showed sufficient weight gain at later examinations. Multiple regression analyses showed that neither cumulative doses nor age had a significant effect on growth when initial weight and height were controlled for. These findings indicate that, for most children, extended treatments with amphetamine or methylphenidate do not have negative effects on growth. However, some children show weight loss during the 1st year of treatment, more often when amphetamine is used. Among those children who may show reduced weight gain, most are above mean weight before treatment begins. PMID- 11869468 TI - Social support in a wired world: use of online mental health forums in Norway. AB - This study explored the use of the four major Norwegian mental-health-related online discussion forums; who participate, why, and what implications use may have. The objective was to provide a basis for proposing relevant research questions and issues for public policy attention. A total of 492 responses to a web-based questionnaire were received. The respondents, predominantly women (78%) in the age range 18--35 years, found forum participation useful for information, and social contact and support. A majority (75%) found it easier to discuss personal problems online than face-to-face, and almost half say they discuss problems online that they do not discuss face-to-face. A majority would not have participated had they not had the option of using a pseudonym. Respondents perceive discussion groups as a supplement rather than a replacement of traditional mental health services, reporting no change in the amount or type of service used. A clear majority want professionals to take an active role in these types of forum. Comments from respondents indicate that forums may have an empowering effect. We believe that online interaction can have unique benefits for people suffering from mental disorders. Professionals will need new knowledge and perceptions of their roles, and public authorities will have to decide their role in influencing the quality of services offered, and the social values conveyed, to those who seek help through the Internet. PMID- 11869469 TI - Re Stig Soderberg's "Personality disorders in parasuicide" in the Nordic journal of psychiatry (2001;55:163-7). PMID- 11869470 TI - Comparisons of lung tumour mortality risk in the Japanese A-bomb survivors and in the Colorado Plateau uranium miners: support for the ICRP lung model. AB - PURPOSES: To estimate the ratio of risks for exposure to radon progeny relative to low-LET radiation based on human lung cancer data, taking account of possible time and age variations in radiation-induced lung cancer risk. MATERIALS AND METHODS: Fitting two sorts of time- and age-adjusted relative risk models to a case-control dataset nested within the Colorado Plateau uranium miner cohort and to the Japanese atomic (A)-bomb survivor mortality data. RESULTS: If all A-bomb survivors are compared with the Colorado data, there are statistically significant (two-sided p < 0.05) differences between the two datasets in the pattern of the variation of relative risk with time after exposure, age at exposure and attained age. The excess relative risk decreases much faster with time, age at exposure and attained age in the Colorado uranium miners than in the Japanese A-bomb survivors. If only male A-bomb survivors are compared with the Colorado data, there are no longer statistically significant differences between the two datasets in the pattern of variation of relative risk with time after exposure, age at exposure or attained age. There are no statistically significant differences between the male and female A-bomb survivors in the speed of reduction of relative risk with time after exposure, age at exposure or attained age, although there are indications of rather faster reduction of relative risk with time and age among male survivors than among female survivors. The implicit risk conversion factor for exposure to radon progeny relative to the A-bomb radiation in the male survivors is 1.8 x 10(-2) Sv WLM(-1) (95% CI 6.1 x10(-3), 1.1 x 10(-1)) using a model with exponential adjustments for the effects of radiation for time since exposure and age at exposure, and 1.9 x 10(-2) Sv WLM( 1) (95% CI 6.2 x 10(-3), 1.6 x 10(-1)) using a model with adjustments for the effects of radiation proportional to powers of time since exposure and attained age. Estimates of the risk conversion factor calculated using variant assumptions as to the definition of lung cancer in the Colorado data, or by excluding miners for whom exposure estimates may be less reliable, are very similar. The absence of information on cigarette smoking in the Japanese A-bomb survivors, and the possibility that this may confound the time trends in radiation-induced lung cancer risk in that cohort, imply that these findings should be interpreted with caution. CONCLUSIONS: There are no statistically significant differences between the male A-bomb survivors data and the Colorado miner data in the pattern of variation of relative risk with time after exposure and age at exposure. The risk conversion factor is very close to the value suggested by the latest ICRP lung model, albeit with substantial uncertainties. PMID- 11869472 TI - Oxic and anoxic enhancement of radiation-mediated toxicity by horseradish peroxidase/indole-3-acetic acid gene therapy. AB - PURPOSE: To evaluate the interaction of horseradish peroxidase (HRP)/indole-3 acetic acid (IAA) gene therapy with therapeutically relevant doses of radiation. MATERIALS AND METHODS: Human T24 bladder and FaDu nasopharyngeal carcinoma cells were transiently transfected with the HRP cDNA using a non-viral delivery method. HRP expression was confirmed by immunostain and enzyme activity assay. The cells were exposed to IAA or the analogue 1-Me-IAA in conjunction with X-rays in air or in anoxic conditions, and cytotoxicity was determined by clonogenic assay. RESULTS: A significant and selective enhancement of radiation-mediated cytotoxicity was observed. Pre-incubation with the prodrugs induced sensitizer enhancement ratios (SER) ranging from 2.6 (0.1mM IAA) to 5.4 (0.5 mM IAA). Radiosensitization was also observed when prodrug exposure was performed immediately after irradiation (SER=2.1-5.6), or in anoxic conditions (SER=3.6). CONCLUSIONS: The use of gene therapy protocols to enhance the effect of ionizing radiation holds promise for anticancer therapy. The data suggest that the combination of HRP/IAA gene therapy with ionizing radiation could present therapeutic advantages in the treatment of solid malignancies, in particular to target the hypoxic population, which has been shown to correlate with poor outcome after radiotherapy. PMID- 11869471 TI - Cytogenetic and cellular events during radiation-induced thymic lymphomagenesis in the p53 heterozygous (+/-) B10 mouse. AB - PURPOSE: Cellular and cytogenetic events in radiation-induced thymic lymphomagenesis were investigated in the p53 heterozygous (+/-) mouse following a single dose of whole-body irradiation. MATERIALS AND METHODS: The loss of the wild-type p53 allele and microsatellite markers of chromosome 11 in thymic lymphomas that developed in the p53 heterozygous (+/-) mouse after irradiation, and the stage at which prelymphoma cells appeared were analysed. RESULTS: The p53 heterozygous mouse developed thymic lymphomas in a dose-dependent manner. The loss of the wild-type p53 allele (loss of heterozygosity; LOH) occurred in almost all thymic lymphomas induced in the irradiated p53 heterozygous mouse. Cytogenetic analysis for the mechanism of LOH strongly suggested that the loss of the wild-type p53 gene in the lymphomas was caused by duplication of the disrupted allele through either homologous recombination or non-disjunctional chromosome duplication. The assay for prelymphoma cells suggested that a critical event in the development of prelymphoma cells occurred at least 3 weeks after irradiation. CONCLUSIONS: The loss of the wild-type p53 gene in thymocytes of the p53 heterozygous mouse may precede the development of prelymphoma cells after irradiation and be a valuable marker of radiation-induced leukemogenesis. PMID- 11869473 TI - Implication of PBP74/mortalin/GRP75 in the radio-adaptive response. AB - PURPOSE: To investigate the relationship between expression of the human peptide binding protein PBP74 and the occurrence of an adaptive response to ionizing radiation. MATERIALS AND METHODS: Human tumour cell lines HT29 and MCF-7 were transfected with a PBP74 or PBP74 antisense construct. For demonstration of an adaptive response, cells lines were irradiated with a conditioning dose of 0.25 Gy cobalt-60 gamma-rays followed by a second dose of 4.0 Gy after an interval of 4.5 h. Response was measured in terms of clonogenic survival. RESULTS: Transfection of a PBP74 plasmid caused transient overexpression of PBP74 mRNA in both cell lines. The optimal dose for the induction of PBP74 in the cell lines investigated was 0.1-0.25Gy and PBP74 induction occurred within 30 min of irradiation. For both cell lines, the adaptive response was repressed when cells were transfected with the anti-PBP plasmid. However, the converse, an enhancement of the adaptive response in cell lines transfected with the PBP74 construct, was seen only for HT29 cells under certain experimental conditions. CONCLUSIONS: The results support the view that while PBP74 is necessary to the adaptive response, it may not by itself be sufficient for the adaptive response to occur. PMID- 11869474 TI - Effect of LET on the yield and quality of chromosomal damage in metaphase cells: a time-course study. AB - PURPOSE: To investigate further the effect of linear energy transfer (LET) on the yield and quality of aberrations at different post-irradiation sampling times. MATERIALS AND METHODS: V79 G(1)-cells were exposed to either 10.6 MeV u-1 Ne ions (360 keV microm-1) or 11.1 MeV u-1 Kr ions (3980 keV microm-1) and chromosomal damage was measured in metaphase cells at several 2-h sampling intervals up to 30 h post-irradiation. To differentiate between cells in the first and second post irradiation cycle, the fluorescence-plus-Giemsa technique was applied. RESULTS: In both experiments, an increase in the yield of aberrant cells as well as the number of aberrations per cell was observed in first- and second-cycle metaphases. Yet, the increase in the number of aberrations per cells was more pronounced for Kr ions and at comparable fluences Kr ions produced more aberrations than Ne ions. Because no sampling time was representative for the whole cell population, the total amount of Ne and Kr ion-induced chromosomal damage was determined by means of a mathematical approach and used for the comparison of data. Furthermore, in accordance with previous studies, LET dependent changes in the spectrum of aberration types were detected, i.e. with increasing LET a higher fraction of chromatid-type aberrations was observed, although cells had been exposed in G1. In addition, more chromosomal breaks and less exchange-type aberrations were found. CONCLUSIONS: The observation that cell cycle progression is related to the amount of aberrations harboured by a cell demonstrates that the routinely applied method to measure aberration frequencies in metaphase cells at only one post-irradiation sampling time will unavoidably result in an under- or overestimation of the cytogenetic effects of particles. Consequently, for a meaningful quantification of chromosomal damage, multiple fixation regimes should be used so that the complete time-course of aberrations can be taken into account. Moreover, to avoid bias, all aberration types should be recorded and included in the analysis since the aberration spectrum changes with LET. PMID- 11869475 TI - Chromosome aberrations detected by FISH and correlation with cell survival after irradiation at various dose-rates and after bromodeoxyuridine radiosensitization. AB - PURPOSE: To determine whether measurement of chromosome aberrations by fluorescence in situ hybridization (FISH) predicts cell survival after irradiation at different dose-rates and after radiosensitization by bromodeoxyurdine (BrdU) in a lung carcinoma cell line. MATERIALS AND METHODS: The human lung carcinoma cell line SW1573 was irradiated at high dose-rate (HDR: 0.8 Gy min-1) or at pulsed low dose-rate (p-LDR: average dose-rate 1 Gy h-1) with or without radiosensitization by bromodeoxyuridine (BrdU). Cell survival was determined by clonogenic assay. Chromosome aberrations (colour junctions) were measured by whole-chromosome FISH of chromosome 2 and 18 and were scored according to the PAINT method. RESULTS: Clear radiosensitization by BrdU was observed both after HDR and p-LDR irradiation. Chromosome 18 was more radiosensitive than chromosome 2. There was a good correlation between induction of colour junctions and cell survival both after HDR and p-LDR irradiation and after radiosensitization by BrdU. CONCLUSIONS: Determination of chromosome aberrations by FISH can predict cell survival after different dose-rates and after radiosensitization by BrdU PMID- 11869476 TI - Radiation-induced peroxidation and fragmentation of lipids in a model membrane. AB - PURPOSE: To ascertain the possibility of fragmentation processes in lipid membranes when acted upon with gamma-radiation under various conditions, and to evaluate fragmentation processes quantitatively in comparison with the lipid peroxidation processes. MATERIALS AND METHODS: Phospholipids as components of multilamellar liposomes exposed to 137Cs gamma-rays at dose-rates of 0.06 and 0.33 Gy s-1 in the dose range 0-18 kGy. Peroxidation products were determined spectrophotometrically and fragmentation products were analysed using thin-layer chromatography and GLC methods. RESULTS: Phosphatidylglycerol containing unsaturated fatty acid residues and free hydroxyl groups underwent both peroxidation and fragmentation when its aqueous dispersions were treated with ionizing radiation. gamma-Radiolysis of multilamellar liposomes led to the formation of peroxidation products, as well as phosphatidic acid and hydroxyacetone, fragmentation products of the initial lipid. Comparable radiation chemical yields of conjugated-diene products and phosphatidic acid indicate that the probabilities of oxidation and fragmentation processes to occur are approximately equal in this case. In contrast to lipid peroxidation, lipid fragmentation is not accelerated but suppressed by oxygen and appears to be a prevailing process in de-aerated solutions. The phosphatidic acid formed as a result of phosphatidylglycerol fragmentation can intensify the lipid peroxidation. CONCLUSIONS: Treatment of lipid membranes with ionizing radiation leads to the destructive processes within both hydrophobic and hydrophilic moiety of lipids. PMID- 11869477 TI - Effects of ionizing radiation on gene expression in cultured rat heart cells. AB - PURPOSE: To determine the in vitro effect of ionizing radiation on TGF-beta1, FGF 2, IL-1beta, atrial natriuretic peptide (ANP) and procollagen types I and III gene expression in three different cell types of rat heart. MATERIALS AND METHODS: Primary cell cultures of myocytes and fibroblasts and cultures of a rat heart endothelial cell line (RHEC) were irradiated with single doses of 2.0, 8.5 or 15 Gy. At different time-points after irradiation (4-336 h), gene expression was analysed using a competitive PCR technique. RESULTS: Irradiation of cultured rat heart cells may lead to temporary changes in expression of the genes studied. Analysis of the radiation response of cultured myocytes, cardiac fibroblasts and rat heart endothelial cells reveals different responses with regard to (1) the dose necessary to evoke changes in mRNA expression, (2) the level of and (3) the duration of the 'induced' response. The changes observed were small and between parallel experiments the onset and time-course of the induced gene expression varied between 4 and 48 h. The average expression of TGF-beta1 mRNA between 4 and 48 h was significantly elevated in endothelial cells after a dose of 2.0 Gy, in fibroblasts after a dose of 8.5 Gy and in myocytes after a dose of 15 Gy. Down regulation of TGF-beta1 mRNA in myocytes was observed after a dose of 8.5 Gy. FGF 2 and procollagen type-I mRNAs were significantly elevated in fibroblasts after a dose of 2.0 Gy. For all three cell types, no effect of dose on the timing or size of the gene expression was observed. CONCLUSIONS: Although irradiation of cultured heart cells influences expression of genes involved in tissue remodelling, the observed differences were too small and too restricted in time and dose to explain the exact role of these cell types in processes leading to radiation-induced cardiac fibrosis. PMID- 11869478 TI - The placental transfer of cerium: experimental studies and estimates of doses to the human fetus from 141Ce and 144Ce. AB - PURPOSE: To measure the transfer of cerium from mother to fetus in experimental animals and estimate doses to the human fetus following intakes of radioisotopes of Ce. MATERIALS AND METHODS: Cerium-141 in chloride solution was administered intravenously to rats at different stages of pregnancy (days 9.5, 12.5 or 18.5), and retention in the embryo/fetus and associated tissues was measured 3 days later in each case. Retention in rat fetal tissues on day 21.5 (shortly before birth) was also measured after administration of 141Ce chloride 1 month prior to conception or 141Ce citrate on day 18.5. Cerium-141 chloride was administered to guinea pigs on day 50 for measurements of fetal retention on day 57 (shortly before birth). RESULTS: Retention of 141Ce in the rat embryo/fetus, measured at 3 days after administration to the mother, increased from about 0.00002% of injected activity per embryo/fetus on day 12.5 to about 0.014% on day 21.5 of gestation. However, the relative concentrations of 141Ce in the embryo/fetus and mother (CF:CM ratio) were between 0.005 and 0.01 in each case. After 141Ce administration prior to conception, retention by the rat fetus on day 21.5 was substantially lower than after short-term administration. Comparison of retention of 141Ce on day 21.5 after administration on day 18.5 as either chloride or citrate showed similar levels in maternal tissues but greater transfer to the fetus (CF:CM ratio of 0.03). Retention in the guinea pig fetus in late gestation at 7 days after administration of (141)Ce chloride was about 0.05% injected activity per fetus, corresponding to a CF:CM ratio of about 0.02. CONCLUSION: These results and other published animal data have been used to specify CF:CM ratios for use in the calculation of doses to the human fetus. The values used were 0.05 for intakes during pregnancy and 0.01 for intakes prior to conception. Doses to the offspring after maternal ingestion of 141Ce or 144Ce are largely due to irradiation from activity in the maternal colon and are insensitive to CF:CM. After inhalation, however, absorption of Ce to blood is much greater and doses to the offspring are dominated by the contribution from activity in the fetus, and therefore dependent on the CF:CM ratio used. PMID- 11869479 TI - Effects of prenatal exposure to the Dutch famine on adult disease in later life: an overview. AB - People who were small at birth have been shown to have an increased risk of CHD and chronic bronchitis in later life. These findings have led to the fetal origins hypothesis that proposes that the fetus adapts to a limited supply of nutrients, and in doing so it permanently alters its physiology and metabolism, which could increase its risk of disease in later life. The Dutch famine--though a historical disaster--provides a unique opportunity to study effects of undernutrition during gestation in humans. People who had been exposed to famine in late or mid gestation had reduced glucose tolerance. Whereas people exposed to famine in early gestation had a more atherogenic lipid profile, somewhat higher fibrinogen concentrations and reduced plasma concentrations of factor VII, a higher BMI and they appeared to have a higher risk of CHD. Though the latter was based on small numbers, as could be expected from the relatively young age of the cohort. Nevertheless, this is the first evidence in humans that maternal undernutrition during gestation is linked with the risk of CHD in later life. Our findings broadly support the hypothesis that chronic diseases originate through adaptations made by the fetus in response to undernutrition. The long-term effects of intrauterine undernutrition, however, depend upon its timing during gestation and on the tissues and systems undergoing critical periods of development at that time. Furthermore, our findings suggest that maternal malnutrition during gestation may permanently affect adult health without affecting the size of the baby at birth. This gives the fetal origins hypothesis a new dimension. It may imply that adaptations that enable the fetus to continue to grow may nevertheless have adverse consequences for health in later life. CHD may be viewed as the price paid for successful adaptations to an adverse intra uterine environment. It also implies that the long-term consequences of improved nutrition of pregnant women will be underestimated if these are solely based on the size of the baby at birth. We need to know more about what an adequate diet for pregnant women might be. In general, women are especially receptive to advice about diet and lifestyle before and during a pregnancy. This should be exploited to improve the health of future generations. PMID- 11869480 TI - Intrauterine growth retardation and puberty in girls. AB - Some, albeit not all studies on the relationship between intrauterine growth retardation (IUGR) and female pubertal development have found an earlier and rapidly progressing puberty as well as concomitant disorders of related functional systems such as polycystic ovary syndrome and short stature. These pubertal changes are part of a growing list of IUGR-related diseases, which includes non-insulin dependent diabetes mellitus and coronary heart disease. A pulsatile release of gonadotropin releasing hormone is thought to be a conditio sinne-qua-non for the initiation of puberty. In the absence of prospective studies on gonadotropin releasing hormone pulse patterns in IUGR-children other markers of pubertal development such as age at menarche have been deployed. From these studies it is not clear, however, whether the findings of an earlier onset of puberty in IUGR-girls merely reflect a more rapid progression of puberty. Both the role for IUGR and the mechanisms behind the onset of puberty are still elusive. Assuming a connection between IUGR and pubertal development, parallels can be drawn between hypotheses on the longterm consequences of IUGR and hypotheses on the initiation of puberty. For example, the somatometer concept proposes a role for fat mass in the initiation of puberty, which is compatible with the hypothesis on non-skeletal catch-up growth after IUGR. The debate on the origins of puberty and the role of IUGR mainly focuses on nature and nurture. Judgmentally, studies in mono- and dizygotic twins discordant for birth weight may be of particular help. PMID- 11869481 TI - Imprinting of genes and the Barker hypothesis. AB - Several common adult diseases appear to be related to impaired fetal growth and this may be caused either by nutritional inadequacies at particular stages of pregnancy or by variation in alleles at specific growth loci. Little is known about the genes involved in the underlying mechanism. This review proposes that at least some of the effects have their origins at imprinted loci, genes that are unusual because they are expressed from only one parental allele. Many imprinted genes are crucial for fetal growth and determine birthweight. They can be disrupted in the early embryo by environmental influences and these disruptions can be inherited through many cell cycles into adult tissues. Their disruption can affect specific organs during fetal development and disruption could affect adult disease in a variety of direct and indirect means. Imprinted genes may be particularly vulnerable to disruption as they are functionally haploid and their expression is regulated by different means from the rest of the genome. Thus many imprinted genes provide plausible candidates for programming adult disease and warrant further study in this context. PMID- 11869482 TI - Birth weight corrected for gestational age is related to the incidence of Down's syndrome pregnancies. AB - Three recent studies reported that early depletion of the primordial follicle pool is likely to be an independent risk factor for Down's syndrome pregnancies. The size of the primordial follicle pool at birth is determined by oogenesis and by the rate of follicle atresia during the intra uterine period. Since intra uterine growth retardation was reported to be associated with a significantly reduced primordial follicle pool at birth, we investigated the possibility of a relation between low birth weight for gestational age and the risk of a Down's syndrome pregnancy. In a case control study, 95 women with a history of a Down's syndrome pregnancy and 85 controls provided information on their own birth weight and length of gestation. Birth weight standard deviation scores, indicating the difference in birth weight from a reference group, were significantly lower in Down's syndrome mothers than in controls. These findings illustrate that the risk of a Down's syndrome pregnancy is related to a low birth weight corrected for gestational age, possibly by a causal relation between intra uterine growth retardation and the size of the primordial follicle pool. PMID- 11869483 TI - The foetal origins of adult disease: interpreting the evidence from twin studies. AB - Twin studies have a contribution to make to the debate concerning the foetal origins of adult disease. Twins are growth retarded compared to singletons and experience post-natal catch-up growth. However, there is no evidence that twins are at increased risk of cardiovascular disease. Studying whether discordance in size at birth within monozygotic twin pairs is predictive of discordance in later life disease should help resolve whether the association between size at birth and later disease is due to common genetic factors. Results from studies of blood pressure in childhood and adult life looking at these within twin effects are far from conclusive. There are, however, methodological problems in the interpretation of these results, not least of which is the relatively small numbers of twin pairs studied. Studies exploring the effect of zygosity and chorion type on later disease provide may provide a useful extension of the research agenda. In summary, twin studies to date have raised more questions about the foetal origins hypothesis than they have resolved. PMID- 11869484 TI - Fetal growth and the fetal origins hypothesis in twins--problems and perspectives. AB - Although there is substantial evidence from studies of singletons that small size at birth is linked with long-term adverse health effects, until recently little was known as to whether these associations extend to twins. A review of published studies suggests that at present there is little consistent evidence that birthsize in twins is associated with increased morbidity or morality. While, these findings may reflect methodological limitations, it is also argued that they arise as a consequence of the substantially different biology of fetal growth in twins. PMID- 11869485 TI - Potential bias regarding birth weight in historical and contemporary twin data bases. AB - In this study we examine the hypothesis that monozygotic (MZ) twins in historical databases are less discordant for birth weight due to negative selection of severely discordant MZ twins. Furthermore, we test the hypothesis that MZ twins are less discordant for birth weight when comparing a volunteer based twin registry with a population based twin registry, due to selective registration. Data were available on 3927 twin pairs from the volunteer Australian Twin Registry born before 1964, 3059 volunteer twin pairs from the Netherlands Twin Register born 1987-1989 and 454 Belgian twin pairs from The East Flanders Prospective Twin Survey born 1987-1989. Intrapair relative birth weight differences (RBWD) were computed for MZ and dizygotic (DZ) twins from each twin registry. Comparing birth weight differences between MZ and DZ twins provides support for the hypothesis that MZ twins are subject to a negative selection in historical databases. Furthermore, Australian MZ twins have a lower RBWD compared to Dutch MZ twins when corrected for the RBWD of Australian and Dutch DZ twins, indicating circumstances which only affect MZ twins. Our hypothesis that MZ twins are less discordant for birth weight in a volunteer based twin registry compared to a population based twin registry had to be rejected. We suggest that investigators using historical databases to test the fetal origins hypothesis should be aware of this increased likelihood of selective exclusion of individuals with extreme morphometric parameters at time of birth. PMID- 11869486 TI - Intra-uterine and genetic influences on the relationship between size at birth and height in later life: analysis in twins. AB - Epidemiological studies have consistently shown a positive association between size at birth (i.e. birth weight or birth length) and height in children, adolescents and adults. To examine whether this association is explained by genetic or nongenetic (intra-uterine) factors, we investigated birth weight, birth length and height in 60 dizygotic and 68 monozygotic adolescent twin pairs still living with their parents. Birth weight of the twins was obtained from their mothers. Height was measured in a standardised way. The mean age was 17+/ 1.7 years for the dizygotic twins and 16+/-1.8 years for the monozygotic twins. Both dizygotic and monozygotic twins with the lowest birth weight from each pair had a height that was lower compared to their co-twins with the highest birth weight (dizygotic twins: 172.2+/-7.9 vs. 173.8+/-9.4 cm [p = 0.05]; monozygotic twins: 171.1+/-9.4 vs. 171.8+/-9.5 cm [p = 0.01]). Similarly, both dizygotic and monozygotic twins with the shortest birth length from each pair had a height that was lower compared to their co-twins with the longest birth length (dizygotic twins: 172.3+/-7.9 vs. 174.9plus minus9.7 cm [p < 0.05]; monozygotic twins: 168.9+/-10.6 vs. 169.9+/-10.2 cm [p < 0.01]). In addition, intra-pair differences in birth weight and birth length were significantly associated with differences in height in both dizygotic twins (regression coefficient: 4.3 cm/kg [95% confidence interval: 1.0 to 7.5] and 0.96 cm/cm [0.17 to 1.74], respectively) and monozygotic twins (2.8 cm/kg [1.4 to 4.1] and 0.73 cm/cm [0.40 to 1.06], respectively). These associations were stronger in dizygotic than in monozygotic twins, but this difference was not statistically significant (for birth weight p = 0.4; and for birth length p = 0.6). However, genetic model fitting indicated that models incorporating a genetic source of the covariance gave a better description of the observed association of birth weight and length with height in later life than models not incorporating this genetic source. The results were similar for data on adult height after 12 years of follow-up in a subgroup of these twin pairs. These data suggest that the association between size at birth and height in later life is influenced by non-genetic intra-uterine and by genetic factors. PMID- 11869487 TI - Cardiovascular mortality in twins and the fetal origins hypothesis. AB - The intrauterine growth patterns for twins are characterized by normal development during the first two trimesters and reduced growth during the third trimester. According to the fetal origins hypothesis this growth pattern is associated with risk factors for cardiovascular morbidity and mortality. We studied cause-specific mortality of 19,986 Danish twin individuals from the birth cohorts 1870-1930 followed from 1952 through 1993. Despite the large sample size and follow-up period we were not able to detect any difference between twins and the general population with regard to all-cause mortality or cardiovascular mortality. Hence, the intrauterine growth retardation experienced by twins does not result in any "fetal programming" of cardiovascular diseases. There is still an important role for twins (and other sibs) to play in the testing of the fetal origins hypothesis, namely in studies of intra-pair differences, which can assess the role of genetic confounding in the association between fetal growth and later health outcome. PMID- 11869489 TI - The influence of zygosity and chorion type on fat distribution in young adult twins consequences for twin studies. AB - An adverse intra-uterine environment has been associated with abdominal fat distribution in singletons. Twins often have a low birth weight and a short gestation. Therefore, they may have an increased risk to develop abdominal obesity. Furthermore, monozygotic monochorionic twins (MZ MC) have a larger intra pair birth weight difference compared to monozygotic dichorionic twins (MZ DC). If adult anthropometry is programmed in utero, this may affect the intra-pair correlations in adulthood and, consequently, also the results from the classic twin method to estimate genetic and environmental influences. In the present study, we compared the absolute values, the intra-pair differences, and the intra pair correlations of body mass, height, BMI, and abdominal fat distribution of 424 MZ MC, MZ DC and dizygotic (DZ) twin pairs (aged 18-34 yrs). DZ, MZ DC and MZ MC twins did not differ for most anthropometric characteristics. Only MZ women tended (p = 0.03) to accumulate more abdominal fat compared to DZ twins. Overall, the contribution of zygosity and chorion type to adult anthropometry was rather low (< or = 1.7%). Although the intra-pair birth weight difference of MZ MC pairs (10.5% in men, 12.3% in women) was significantly larger compared to that of MZ DC pairs (6.9% and 9.2% resp.), the intra-pair differences in adult anthropometry were similar for both MZ twin types. Also the intra-pair correlations of MZ MC and MZ DC pairs were strikingly alike, suggesting no significant influence of the prenatal environment on adult concordance. In conclusion, the substantial difference in the prenatal environment of MZ MC and MZ DC twins did not result in a difference in intra-pair concordance of adult anthropometry and fat distribution. Therefore, we suggest that the chorion type of MZ twins does not bias the twin design and the estimation of the genetic contribution to adult anthropometry. PMID- 11869488 TI - Glucose and insulin metabolism in twins: influence of zygosity and birth weight. AB - Several epidemiological and metabolic studies have demonstrated an impact of the intrauterine environment on the development of disease in adult life, including Type 2 diabetes and glucose intolerance. Our finding of lower birth weights among monozygotic diabetic twins compared to their non-diabetic genetically identical co-twins confirms this association and, furthermore, eliminates the possibility that the association could be explained solely by common genes leading to both impaired intrauterine growth and increased risk of Type 2 diabetes. Due to an often shared placenta monozygotic twins may experience a more adverse intrauterine environment compared to dizygotic twins and may therefore be more prone to develop various metabolic abnormalities. Our findings of a higher glucose and insulin profile after oral glucose ingestion, and recently lower insulin-stimulated glucose uptake--indicating glucose intolerance and insulin resistance--among monozygotic compared to dizygotic twins may to some extent question the validity of classical twin studies in diabetes research where equal environmental influences in monozygotic and dizygotic twins is assumed. The potential role of an adverse intrauterine environment in causing Type 2 diabetes in humans, may to some degree alter our conception of the twin model in diabetes research including the interpretation of aetiological conclusions reached in previous classical twin studies of diabetes. However, our present knowledge is far too insufficient to discard the results from classical twin studies concerning the relative role of genes versus environment for the development of diabetes and its metabolic effects. PMID- 11869490 TI - Genetic and non-genetic factors affecting birth-weight and adult Body Mass Index. AB - Birthweight affects neonatal mortality and morbidity and has been used as a marker of foetal undernutrition in studies of prenatal effects on adult characteristics. It is potentially influenced by genetic and environmental influences on the mother, and effects of foetal genotype, which is partially derived from the maternal genotype. Interpretations of variation in birthweight and associated characteristics as being due to prenatal environment ignore other possible modes of materno-foetal transmission. Subjects were adult twins recruited through the Australian Twin Registry, aged 17 to 87 years, and the sample comprised 1820 men and 4048 women. Twins reported their own birthweight as part of a health questionnaire. Body Mass Index (BMI) was calculated from self reports of height and weight. Correlations between co-twins' birthweights were high for both monozygotic (r = 0.77) and dizygotic (r = 0.67) pairs, leading to substantial estimates of shared environmental effects (56% of variance) with significant additive genetic (23%) and non-shared environmental (21%) components. Adult BMI was mainly influenced by genetic factors, both additive (36% of variance) and nonadditive (35%). The correlation between birthweight and BMI was positive, in that heavier babies became on average more obese adults. A bivariate model of birthweight and adult BMI showed significant positive genetic (r(g) = 0.16, p = 0.005) and environmental (r(e) = 0.08, p = 0.000011) correlations. Intra-uterine environmental or perinatal influences shared by cotwins exercise a strong influence on birthweight, but the factors which affect both birthweight and adult BMI are partly genetic and partly non-shared environmental. PMID- 11869491 TI - Early changes in vascular dynamics in relation to twin-twin transfusion syndrome. AB - A clearer understanding of the early determinants of normal and abnormal vascular development is pivotal in order to identify those at increased risk of later vascular disease, and perhaps to prevent it by early intervention. Measurement of pulse wave velocity(PWV) has been used in the postnatal evaluation of the monochorionic(MC) twins. They are genetically identical and those with twin-twin transfusion syndrome(TTTS) provide an ideal natural model in whom to study the influence of differing haemodynamic stresses on the developing vascular tree. We investigated firstly whether surviving twin pairs with TTTS have altered arterial distensibility in childhood by comparing PWV in the radial arteries of surviving MC twin pairs with TTTS and in two control groups, one cohort of MC twins without TTTS and another dichorionic group (DC) Secondly, we tested a cohort of TTTS twin pair survivors treated with laser photocoagulation. The co-twin pairs in the group managed palliatively with amnioreduction showed increased PWV in the donor and reduced PWV in the recipient twins. This was neither seen in the laser treated, nor in the control groups. Our studies suggest that a period of haemodynamic imbalance gives rise to changes in a muscular conduit artery that persist at least into infancy and it seems that by correcting the abnormal haemodynamics relatively soon after the disease process had begun, the alterations in elasticity are prevented. These studies are the first to demonstrate fetal programming of the vascular bed in humans, and prevention or reversal of this programming by an intervention in mid-gestation. PMID- 11869492 TI - Association of birth weight and current body size to blood pressure in female twins. AB - It has been proposed that low birth weight is associated with high levels of blood pressure in later life. The aim of this study was to assess the relationship of blood pressure to birth weight and current body size during growth and adulthood. A total of 711 female multiple births, with one group of 244 in their growth phase mean age 12.0 (2.3)(SD) years and the other of 467 adults (mean age 35.2 (12.6) years), had height, weight and both systolic (SBP) and diastolic (DBP) blood pressures measured, and self-reported their birth weight. Regression analyses were performed to assess the cross-sectional and within-pair associations of blood pressure to birth weight, with and without adjustments for current body size. Within-pair analysis was based on 296 twin pairs. Cross-sectionally, a reduction in birth weight of 1 kg was associated with 2 to 3 mm Hg higher age-adjusted SBP, which was of marginal significance and explained about 2% of the population variance. Adjustment for body mass index did not significantly change this association. Within-pair analyses found no association between birth weight and SBP or DBP,even after adjusting for current body size. After age, current body size was the strongest predictor of systolic BP. The weak association of blood pressure to birth weight cross-sectionally is of interest, but any within-pair effect of birth weight on blood pressure must be minimal compared with the effect of current body size. PMID- 11869493 TI - Comparing blood pressure of twins and their singleton siblings: being a twin does not affect adult blood pressure. AB - The hypothesis was tested that monozygotic (MZ) and dizygotic (DZ) twins, with their lower average birth weight, have higher adult blood pressure than their singleton brothers or sisters. From the Netherlands Twin Registry, 261 twin families were recruited from a young adult and an older adult cohort with mean ages of 26.2 and 50.4 respectively. These families yielded 204 MZ twins with 71 singleton siblings and 271 DZ twins with 103 of their singleton siblings. Anti hypertensive medication use of these 649 participants was assessed twice with a two-year interval. Resting blood pressure was measured thrice during a standardized laboratory protocol. In spite of a significant difference in birth weight (1036 gram), no differences were found in anti-hypertensive medication use at both time points between twins and singletons nor between their resting laboratory diastolic or systolic blood pressure. These results applied to each gender and to both age cohorts. Limiting the analyses to matched twin-sibling pairs of the same families and taking current weight and height into account did not change the results; no evidence was found for a twin-singleton difference. It was concluded that estimates of genetic and environmental contributions to blood pressure deriving from twin studies do not appear to be biased and may be generalized to singletons. Our results suggest that the lower birth weight in twins does not reflect the intrauterine disadvantage described by the Barker hypothesis. PMID- 11869495 TI - Birthweight and body mass index in young adulthood: the Swedish young male twins study. AB - Many studies have found an inverse association between fetal growth and cardiovascular disease related to the metabolic syndrome in adulthood. Nevertheless, the relative importance of genetics and the intrauterine environment remain unclear. The objective of the study was to test the fetal origins hypothesis and the fetal insulin resistance hypothesis by studying the impact of fetal growth on Body Mass Index (BMI) in young adulthood. In a nationwide cohort study, the Swedish Medical Birth Register for the years 1973 1979 was linked with the Military Service Conscription Register for 1990-1999. In 1998 a questionnaire was mailed to all male twins, included in the two registers, who were alive and still resident in Sweden. The study covers the 923 male twin pairs for which full data were available. Mixed linear models were used to estimate within-pair and between-pair differences in birthweight and their relations to BMI. A weak positive association was found among the monozygotic twins for the withinpair difference in birthweight and BMI. No significant association was found among the monozygotic for the between-pair difference in birthweight and BMI. No significant associations were found for dizygotic twins. These findings do not seem to support either the fetal programming hypothesis or the fetal insulin resistance hypothesis. PMID- 11869494 TI - Perinatal risk factors for hay fever--a study among 2550 Finnish twin families. AB - Previous studies have suggested that perinatal factors influence the risk for asthma but population studies on perinatal factors and risk for hay fever are few. We studied the effect of perinatal factors on the risk for hay fever among adolescent twins by a questionnaire study involving five consecutive nation-wide birth cohorts of 16-year-old twins and their parents. The risk for parent reported, doctor-diagnosed hay fever in the adolescents associated with several perinatal characteristics was assessed with logistic regression analysis among individuals and by a discordant pair analysis. In the univariate analysis of the birth factors, the risk for hay fever increased with increasing birth weight (p for trend = 0.048, OR for those > or = 3000 g 1.35, 95% CI 0.91-2.02 compared to those < 2000 g) and gestational age (p for trend = 0.04, OR for those born after 40 weeks of gestation 2.24, 95% CI 1.03-4.86, compared to those born before 33 weeks of gestation) and was lower in those subjects hospitalised in the neonatal period (OR 0.74, 95% CI 0.58-0.93). Because of significant interactions between parental hay fever status and birth factors (ponderal index, p = 0.03 and maternal age p = 0.04), stratified analysis were performed. The positive association between birth weight and hay fever was most obvious among adolescents with no parental history of hay fever (p for trend = 0.03). Similar, though not significant, trends were found with other birth factors among these families, whereas no such trend was found among adolescents with parental hay fever, suggesting that gestational maturity increases the risk for hay fever in the absence of genetic predisposition. However, of the perinatal factors only neonatal hospitalisation (OR 0.75, 95% CI 0.59-0.96) remained a significant risk factor for the development of hay fever, when adjusted for non-perinatal factors. PMID- 11869496 TI - Intergenerational effect of an adverse intrauterine environment on perturbation of glucose metabolism. AB - Human epidemiological and animal studies have revealed the late consequences of malnutrition during gestation and early life on the health of the offspring. These studies have highlighted the inverse relationship between birth weight and the incidence of insulin resistance and type 2 diabetes later in life. The aim of this paper is to review the different means of achieving foetal malnutrition and its consequences even for a next generation, in animal models and to identify key area for further research. We address the impact of two models of maternal malnutrition (protein restriction and caloric restriction) as well as the impact of maternal diabetes, the three maternal conditions leading to perturbed foetal nutritional environment. Particular emphasis is given to the endocrine pancreas and the insulin sensitive tissues. More specifically, alterations of the foetal nutritional environment perturb the development of the endocrine pancreas and target the ss cell mass at birth. Some adaptations later in life may take place but stress situations such as pregnancy and ageing precipitate the animals to glucose intolerance and insulin resistance. Even the next generation features alterations in the development of the endocrine pancreas. Some mechanisms by which the foetal ss cell mass is altered are approached in this review and specific attention is paid to the amino acid profile. The preventive role of taurine is discussed. PMID- 11869497 TI - Health of human and livestock conceived by assisted reproduction. AB - Assisted reproduction is used to resolve infertility problems in human and in breeding programs to generate livestock. Except for gestation length and birth weight, perinatal outcome of children conceived by In Vitro Fertilization is similar to that of spontaneously conceived children. However, large offspring syndrome observed after In Vitro Production in livestock is quite alarming. The distinct parts of assisted reproduction (oocyte maturation, fertilization and culture) have been found to contribute to abnormal fetal growth and development. Genomic imprinting is suggested to be involved in the induction of the aberrant phenotypes observed after assisted reproduction. Furthermore, current knowledge on postnatal health of offspring conceived by assisted reproduction and speculations on potential longterm effects of In Vitro Fertilization will be described. PMID- 11869498 TI - Implant prosthodontic management of anterior partial edentulism: long-term follow up of a prospective study. AB - OBJECTIVE: This paper reports on the long-term outcome of patients with Kennedy Class IV partial edentulism treated in the Implant Prosthodontic Unit (IPU) at the University of Toronto, Toronto, Ontario. METHODS: The information for this paper was gathered from the charts of the first 30 consecutive, partially edentulous patients treated at the IPU. These patients all had Class IV edentulism and formed part of the original prospective clinical studies that were initiated in 1983. The patients' dental history suggested maladaptive experiences with traditional removable prostheses or a reluctance to have intact or quasi intact teeth prepared as retainers for fixed prostheses. Fifteen men and 15 women treated with 94 Br nemark dental implants, supporting 34 prostheses, were followed until June 2000 (25 patients) or until they were lost to follow-up (5 patients). The multiple missing teeth occurred in 19 maxillae and 15 mandibles. RESULTS: The original prosthodontic treatments were intended to result in 33 fixed partial prostheses and 1 overdenture. At the time of this report, 25 patients with 86 implants supporting 31 fixed prostheses and 3 overdentures had been followed for an average of 12 years (range 7 16 years). The overall survival of implants was 92%. The difference between men (94%) and women (89%) was not statistically significant. CONCLUSIONS: This report is an interim update on an ongoing long-term prospective study. The results so far demonstrate a high survival rate for Br nemark implants supporting tissue-integrated prostheses for the management of anterior partial edentulism. PMID- 11869499 TI - Outcomes of implant prosthodontic treatment in older adults. AB - Older adults are expected to account for an increasingly disproportionate number of individuals needing oral implant prostheses. However, this biotechnology was initially studied for predominantly middle-aged edentulous patients, not elderly people. High rates of success and minimal crestal bone loss have been reported for oral implants mainly in this group. The results of studies at the University of Toronto now clearly support earlier reports that older adults respond to oral implants in the same manner as younger adults, despite their tendency for systemic illness, including osteoporosis. However, unfavourable jawbone quantity and quality, particularly atrophy of the maxilla, impaired implant success. Furthermore, placement of implants in sites that had been edentulous for shorter periods was associated with greater crestal bone loss, a finding that may have implications for younger adults undergoing such treatment. The major decision making challenge in managing depleted dentitions and complete edentulism in an aging society now lies in differentiating the treatment outcomes, especially patient-mediated assessments (including economic analyses), of the various prosthodontic options available for older adults. PMID- 11869500 TI - Implant prosthodontics in medically challenged patients: the University of Toronto experience. AB - A series of prospective studies started in the mid-1980s at the University of Toronto have provided evidence of the efficacy and effectiveness of implants in the treatment of the fully and partially edentulous patients. These studies have focused primarily on treatment outcomes at the surgical and prosthodontic levels, with an overall failure rate of 7.7% over a 20-year period. Because a considerable proportion of these failures (4.2%) occurred before insertion of the prosthesis, and because osseointegration is essentially a wound-healing process, factors that interfere with healing, including systemic conditions, may contribute to implant failure. This paper reviews studies on the impact of selected systemic conditions, including osteoporosis, cardiovascular diseases, diabetes mellitus, and hypothyroidism, as well as smoking behaviour, on the success or survival of oral implants in patients treated in the Implant Prosthodontic Unit at the University of Toronto. PMID- 11869501 TI - A 5-year prospective study of implant-supported single-tooth replacements. AB - OBJECTIVE: Because osseointegration has been successful in the management of completely edentulous patients, it is tempting to extrapolate these results and infer the success of single-tooth replacement. Yet there are major clinical differences between edentulous and partially edentulous patients. This prospective study is a follow-up to one started at the University of Toronto in 1986. The purpose of this study was to continue longitudinal assessment of implant-supported single-tooth replacements. METHODS: The original study comprised 42 consecutively treated patients with a total of 49 implants. The patient group consisted of all University of Toronto patients treated with single Br nemark implants whose treatment had been completed more than 5 years previously (i.e., before 1994). No exclusion criteria applied. One implant was not osseointegrated at the time of stage 2 surgery, and 6 patients with reportedly successful osseointegrated implants were not available for recall. For the preparation of this report, 30 of the remaining 42 implants were assessed during recall examinations. Assessment of success was based on published criteria. In addition, soft-tissue appearance, implant immobility, occlusal contacts in centric occlusion and excursions, proximal contacts, tightness of crown and abutment screws, and patients' responses on satisfaction questionnaires were evaluated. RESULTS: The criteria defining success of treatment in implant prosthodontics were met by all 30 of the single-tooth implants, which had been in place for 5 or more years. Each implant was immobile, and each had a mean vertical bone reduction of less than 0.2 mm annually. CONCLUSION: Stable long term results can be achieved with single Branemark implant-supported crowns. PMID- 11869502 TI - Implant prosthodontic management of posterior partial edentulism: long-term follow-up of a prospective study. AB - OBJECTIVE: This paper reports on the long-term outcome of implant-supported posterior-zone prostheses in the first 35 consecutive, partially edentulous patients treated in the Implant Prosthodontic Unit (IPU) at the University of Toronto, Toronto, Ontario. METHODS: A total of 106 Branemark dental implants were placed in 46 posterior edentulous spans in 35 patients for the management of multiple missing teeth; the patients were followed prospectively. Treatment planning principles involved a minimum of 2 or 3 implants at each edentulous site and scrupulous occlusal prosthodontic designs. RESULTS: The overall survival of posterior implants was 94%. No factors in the patients' history adversely affected implant survival. CONCLUSIONS: This clinical update suggests that the use of Branemark implants in the rehabilitation of patients who are partially edentulous in the posterior zone is highly effective and that survival of the implants is excellent. PMID- 11869503 TI - Business planning for university health science programs: a case study. AB - Many publicly funded education programs and organizations have developed business plans to enhance accountability. In the case of the Department of Dentistry at the University of Alberta, the main impetus for business planning was a persistent deficit in the annual operating fund since a merger of a stand-alone dental faculty with the Faculty of Medicine. The main challenges were to balance revenues with expenditures, to reduce expenditures without compromising quality of teaching, service delivery and research, to maintain adequate funding to ensure future competitiveness, and to repay the accumulated debt owed to the university. The business plan comprises key strategies in the areas of education, clinical practice and service, and research. One of the strategies for education was to start a BSc program in dental hygiene, which was accomplished in September 2000. In clinical practice, a key strategy was implementation of a clinic operations fee, which also occurred in September 2000. This student fee helps to offset the cost of clinical practice. In research, a key strategy has been to strengthen our emphasis on prevention technologies. In completing the business plan, we learned the importance of identifying clear goals and ensuring that the goals are reasonable and achievable; gaining access to high-quality data to support planning; and nurturing existing positive relationships with external stakeholders such as the provincial government and professional associations. PMID- 11869504 TI - [The patho-morphology and biological behavior of repeatedly recurrent liposarcoma in retroperitoneum]. AB - OBJECTIVE: To investigate the patho-morphological features and biological behavior of repeatedly recurrent liposarcomas in retroperitoneum as well as the differential diagnosis with other malignant mesenchymal tumors. METHODS: Totally 10 patients accompanied with detailed clinical data. Except HE staining, immunohistochemical assays for S-100 actin, vimentin and myoglobin were also used for 32 specimens. RESULTS: 6/8 cases of the primary and 13/24 cases of the recurrent tumors of this group were diagnosed as sclerosing liposarcoma with spindle cell proliferation arranged in fascicles and whorls. Ten retroperitoneal tumor recurred within 20 months to 10 years, and among them, distant metastasis obtained only in one case. The histologic subtypes of recurrent and metastasized tumors became varied in 5 cases. CONCLUSION: Sclerosing liposarcoma was the most common histological subtype, see in retroperitoneal liposarcoma, local recurrence frequent, but distant metastasis rare. Surgical removal of the recurrent tumors might prolong the survival expectation of these patients. PMID- 11869505 TI - [Diagnosis and differential diagnosis of primary carcinoma of female peritoneum]. AB - OBJECTIVE: Histogenesis and morphological features of 6 primary carcinomas of female peritoneum were studied. METHODS: H&E staining, histochemical stainings and immunohistochemical stainings were used. RESULTS: Among 6 primary carcinomas there were 4 cases of serous papillary carcinomas, 1 case of endometrioid carcinoma and 1 case of transitional cell carcinoma. CONCLUSION: Primary carcinomas of the female peritoneum were histologically identical to that of the similar epithelial carcinomas arising from the ovaries. The diagnosis was made only when the ovaries are not involved and without no any evidence of similar epithelial carcinoma obtained in the ovaries, uterus or urinary organs. These tumors are considered arising from the secondary Mullerian system. D-PAS and CEA staining are helpful as the accessory criteria for the differential diagnosis between primary carcinoma of the peritoneum and malignant mesothelioma. PMID- 11869506 TI - [A clinico-pathological study of neuroendocrine tumors in stomach]. AB - OBJECTIVE: To study the pathological classification of gastric neuroendocrine tumors and its clinico-pathological significance. METHODS: Paraffin sections of totally 52 gastric neuroendocrine tumors including 42 carcinoid tumors were studied with immunohistochemical technique, which involved 9 endocrine markers of hormones antibodies and electronic microscopy for investigating the endocrine cells and the contiguous gastric mucosa of the neuroendocrine tumors. RESULTS: The 52 gastric neuroendocrine tumors were divided into three types: Type I, carcinoid, associated with atrophic gastritis, altogether 26 cases. Tumor extension was limited in the mucosa or submucosa, accompanying with hypergastrinemia and G cell hyperplasia. This type is consistently preceded by and associated with generalized proliferation of endocrine cells in the mucosa at fundus. Type II, carcinoid of sporadic type, totally 16 cases, not associating with hypergastrinemia, was more aggressive. Type III, Neuroendocrine carcinomas (10 cases), were highly aggressive tumors. CONCLUSION: A correct identification of different types of gastric endocrine tumors is important and implicit for the treatment and prognosis of neuroendocrine tumors. PMID- 11869507 TI - [Research on the oncogenesis of cervical epithelial cells co-induced by human papillomavirus and human cytomegalovirus]. AB - OBJECTIVE: The carcinogenesis of the human cervical carcinoma is known closely associated with human papillomavirus (HPV). The purpose of this article is to identify whether another sexually transmitted factor, human cytomegalovirus (HCMV) plays as a co-factor role in the carcinogenesis. METHODS: Human cervical epithelial cells immortalized by HPV16 were transfected with IE gene of HCMV. The condition and efficiency of IE gene were examined. RESULTS: It was showed that the cervical epithelial cells could be completely transformed into malignant by HPV 16 and IE gene of HCMV and tumor was formed by injecting the transformed cells in nude mice. CONCLUSION: The co-operation effect of HPV and HCMV auailed the oncogenesis of human cervical carcinoma and moreover, the cellular and molecular biological mechanism was discussed. PMID- 11869508 TI - [A preliminary study of loss of heterozygosity on chromosome 1p in primary hepatocellular carcinoma]. AB - OBJECTIVE: Ten (10) loci on chromosome 1p were analyzed to detect LOH in 38 cases of hepatocellular carcinomas (HCC) in order to locate the deletion area and the possible deleted tumor suppressor gene (TSG) in HCC. METHODS: PCR based microsatellite instability analysis were used to detect LOH on chromosome 1p31 and 1p35-p36 in HCC. RESULTS: LOH were detected on chromosome 1p in 84.2% of HCC. High frequency of LOH (> 40%) were occurred at locus D1S186 on 1p31, locus D1S482 on 1p35 and loci D1S243, D1S160, D1S165 and D1S170 on 1p36. CONCLUSION: LOH occurred mostly on chromosome 1p31 and 1p35-p36 in HCC. There may be more than two TSGs which associated with development of HCC on chromosome 1p31 and 1p35 p36. PMID- 11869509 TI - [HBxAg enhanced p53 protein accumulation in hepatoma cells]. AB - OBJECTIVE: In order to determine the interaction between HBxAg and p53 protein and its role in hepatocarcinogenesis. METHODS: Cotransfection was performed using reporter gene, chloramphenical acetyltransferase (CAT), p53 and HBVx gene, and dexamethasone inducible HBx plasmid pMAMHBx was constructed for the experiment. Human hepatoma cell line Hep3B was used as the target cell. The activity of HBxAg and p53 was detected by CAT assay and immunohistochemical staining after transient transfection. RESULTS: HBxAg could increase the accumulation of p53 protein in cells and enhance CAT expression. Immunohistochemical staining showed that p53 protein bound to HBxAg in the cytoplasm and formed a complex causing a partial block of the process of p53 during passing through the nuclear membrane from the cytoplasm. CONCLUSION: Data strongly suggested that HBxAg is capable of binding p53 and forms a protein-protein complex which might reduce or inactivate p53's antiproliferative activity and plays an important role in the pathogenesis of HBV-associated hepatocellular carcinoma. PMID- 11869510 TI - [Expression of transforming growth factor alpha and its relationship with HBV infection in hepatocellular carcinomas]. AB - OBJECTIVE: To assess the expression of TGF alpha in hepatocellular carcinoma and its relationship with HBV infection. METHODS: Using in situ hybridization and streptavidin-peroxidase immunohistochemistry to detect TGF alpha mRNA, TGF alpha protein and HBV DNA in 53 cases of HCC and 14 cases of controls. RESULTS: Positive rates of TGF alpha mRNA and TGF alpha protein in carcinoma tissue were 30.2% and 73.6% respectively, which were higher than in normal controls (P < 0.05). The expression of TGF alpha mRNA and TGF alpha protein was increased significantly in dysplastic liver cells (DLC) locating aside from the tumor tissue in comparing with those of non-dysplastic liver cells (NDLC) (P < 0.05). Positive rates of HBV DNA in carcinoma tissue and paratumorous liver tissue were 56.6% and 60.4% respectively, which were higher than in the normal controls (P < 0.001). Positive rate of HBV DNA in the nuclei of tumor tissues, tissue of the dysplastic areas and non-dysplastic areas was lowering down successively (P < 0.05). There was a significant relationship between expression of TGF alpha mRNA and TGF alpha protein in the carcinoma tissues and HBV DNA in the tumorous and its surrounding tissues respectively. CONCLUSIONS: Expression of TGF alpha in carcinoma tissues was closely associated with the existence of HBV DNA and TGF alpha may participate in the early stage of liver carcinogenesis. The increased expression of HBV DNA in nuclei may serve as an important morphology marker in distinguishing carcinoma from DLC and NDLC. PMID- 11869511 TI - [The mechanism of p16 gene inactivation in human bladder transitional cell carcinoma]. AB - OBJECTIVE: To figure out the mechanism of p16 gene inactivation in human bladder transitional cell carcinoma (BTCC). METHODS: p16 gene was analyzed for genetic alterations by Southern blot analysis, PCR-SSCP and DNA methylation analysis in 52 cases of transitional cell carcinoma of bladder. RESULTS: p16 gene deletion was found in 11 of 52 (21.15%) BTCCs, only one tumor showed point mutation on p16 exon 2, nineteen cases of BTCCs (36.53%) showed p16 gene 5' CpG island methylation. CONCLUSION: There was a statistically significant association between p16 gene deletion and early stage, well differentiated tumors. Additionally, a statistically significant correlation was also noticed between p16 gene 5' CpG island methylation and tumors in late stage with poor cell differentiation. PMID- 11869512 TI - [Molecular cloning of hTRT catalytic domain and its expression in tumors]. AB - OBJECTIVE: To study the expression of hTRT gene in tumors and its significance in cancer diagnosis. METHODS: The expression of hTRT in HeLa cells and PG cells was estimated by RT-PCR and the hTRT cDNA encoding the catalytic domain from HeLa cells were cloned and sequenced. Using in situ hybridization (ISH) techniques, the expression of hTRT mRNA in 54 human tumors were observed and compared with the expression of hTR. RESULTS: hTRT was detected in both HeLa and PG cells. Sequencing showed that the cloned hTRT catalytic domain cDNA was identical with that reported in the literature. hTRT and hTR expressions were detected in 38/46 and 40/46 malignant tumors respectively, while both hTRT and hTR were not detected in 8 benign tumors. CONCLUSION: hTRT was detected in both HeLa and PG cancer cells. hTRT catalytic domains were highly conservative. The expression of hTRT and hTR were both high in malignant tumors and they were strongly correlated. Our results suggest that detection of hTRT was valuable to clinical oncology diagnosis. PMID- 11869513 TI - [Detection of Epstein-Barr virus, cell proliferation and apoptosis in thymic tumors]. AB - OBJECTIVE: To investigate the incidence of thymic tumors in Guangzhou, its association with EBV infection and the cell proliferation and apoptosis characteristics of these tumors. METHODS: EBV encoded small RNAs (EBERs) were detected in 43 thymic tumors and 7 thymic hyperplasia samples by use of in situ hybridization, EBNA-1, LMP-1, PCNA, bcl-2 and p53 were examined using immunohistochemical methods, cell apoptosis was evaluated by TUNEL (TdT-mediated dUTP-X nick end labelling) in situ cell death detection kit. RESULTS: 1. Thymic diseases only accounted for 0.057% of biopsies in the Guangzhou area. Thymic tumor and hyperplasia accounted for 74.7% and 18.6% in thymic diseases respectively. Thymic lymphoepithelimoa-like carcinoma (LELC) occurred in 2.7% of thymic tumors. 2. EBV encoded products (EBERs, EBNA-1 and LMP-1) were not detected in 50 thymic disease specimens. 3. Marked proliferation (3+-4+) grade thymomas (0/18 cases) was much lower than that of thymic carcinomas (10/20 cases), and the marked apoptotic (2+-4+) grade was higher in thymomas (15/18) than the thymic carcinomas (13/20). 4. The p53 protein accumulation rate in thymic carcinomas (8/20) was significantly higher than that of thymomas (1/18). The apoptotic grade is significantly related to p53 accumulation in thymic epithelial tumors. 5. The bcl-2 over expression could be detected both in thymomas and thymic carcinomas. CONCLUSIONS: 1. Thymic diseases, including thymic LELC, is very rare in Guangzhou and is not closely associated with EBV infection. 2. Thymic carcinomas have a relatively rapid growth rate compared to thymomas. 3. p53 gene mutation may play an important role in the carcinogenesis of thymic carcinoma, and bcl-2 over-expression may also be involved in the evolution of thymic epithelial tumors. 4. Wild type p53 protein may exert an apoptosis inducing function in thymic epithelial tumors, including thymomas and carcinomas, and a proliferation inhibiting function only in thymomas. PMID- 11869514 TI - [Observations on prostate histology in 50 young and middle aged men. Its implication on clinical pathological diagnosis]. AB - OBJECTIVE: To investigate the morphology and the relationship between the prostate and the adjacent structures in order to clarify its significance in histopathological diagnosis. METHODS: By means of light microscopy, and in part by electron microscopy and immunohistochemistry, the histology of 50 normal prostates were observed. RESULTS: The parenchymal component can be divided into four zones, i.e. peripheral zone, central zone, transitional zone and periurethral gland region, each characterized by its own anatomical and histological features. CONCLUSION: This 4 zone division of prostate has more clinical significance than the traditional division of the prostate. A good knowledge of the normal structure of the prostate has great practical significance in pathologic diagnosis. PMID- 11869515 TI - [The detection of microsatellite instability in carcinoma and adenoma of large bowel and its significance]. AB - OBJECTIVE: To explore the role of microsatellite instability (MSI) in colorectal carcinogenesis and the relationship between MSI, proliferation activity and prognosis. METHODS: PCR-SSLP and immunohistochemistry methods were used to detect MSI and the expression of PCNA in 56 cases of carcinomas, 9 cases of adenomas and 6 cases of adenomas with malignant changes. RESULTS: The total positive rate of MSI were 25/56 cases in colorectal carcinomas. The MSI positive cases were 3/4 in HNPCCs, 22/52 in sporadic colorectal carcinomas, 2/9 in adenomas and 2/6 in adenoma with malignant changes respectively. The PCNA labelling index of MSI positive tumors were significantly lower than that of MSI negative tumors (P < 0.01). The 3 and 5 year survival rates in patients with MSI positive colorectal carcinomas were higher than those of MSI negative tumors. CONCLUSION: MSI may be an early molecular alteration and another molecular mechanism in colorectal carcinogenesis. The MSI positive tumors have low proliferating activity and a better clinical outcome. The detection of MSI might be useful in predicting prognosis of colorectal carcinomas. PMID- 11869516 TI - [The amyloidosis in cardiac conduction system]. AB - OBJECTIVE: To investigate the relationship between arrhythmia, cardiac failure and amyloidosis of cardiac conduction system (CCS). METHODS: CCS from 6 cases with cardiac amyloidosis were sampled using technique designed by the authors. The CCS were studied by Congo red staining and polarization microscopy. RESULTS: Pathological changes could be found in the CCS of these 6 cases. It was serious in sino-atrial node while slight in atrio-ventricular conduction system. The changes in atrio-ventricular conduction system were exacerbated from proximal part (atrio-ventricular node) to distal part (left bundle branch). In the 3 cases with sinus bradycardia and conduction blocking manifested by electrocardiogram, the pathological changes in CCS were more extensive and serious than that of other cases. CONCLUSION: The amyloidosis in CCS is one of the pathological bases of arrhythmia. PMID- 11869517 TI - [A study on homozygous deletion, hypermethylation, mutation and expression of p16 gene in human breast cancer]. AB - OBJECTIVE: To investigate the homozygous deletion (HZD), hypermethylation and mutation of p16 gene in human breast cancer and the relationship between the structural alterations of p16 gene and its expression. METHODS: PCR and PCR methylation assay with silver staining (PCR-MASS) were used to detect HZD and hypermethylation of p16 gene exon 1 in 60 fresh breast cancers and 24 normal breast tissues adjacent to cancer (as control tissues). PCR-SSCP and DNA sequencing were used for the analysis of p16 gene mutation. Moreover, p16 gene and mRNA were also detected in the 60 cases. RESULTS: Of the 60 breast cancers, HZD was found in 7 cases, hypermethylation and mutation were found in 16 and 4 cases respectively, and the difference was statistically significant. The positive rates of p16 protein and mRNA in breast cancer were 28/60 and 39/60 respectively. CONCLUSION: The results demonstrate that several kinds of p16 gene structural changes exist in breast cancer. The structural changes of p16 gene cause abnormal p16 expression, the main mechanisms are hypermethylation, while HZD or mutation are the secondary causes. Abnormal expression of p16 gene then becomes involved in the development and metastasis of breast cancer. PMID- 11869518 TI - [Clinicopathological study of 52 cases of ameloblastoma of jaw]. AB - OBJECTIVE: To observe the relationship between the histologic types, tumor invasion, operation mode and recurrence of ameloblastoma. METHODS: Using WHO classification (1992) for histological typing of odontogenic tumors, 52 cases of ameloblastoma were studied clinicopathologically. RESULTS: The 8 pathological types are: follicle formation type, plexiform type, acanthoma type, basal cell type, granular cell type, desmoplastic ameloblastoma, kerato ameloblastoma and ameloblastoma accompanied by bone formation. Recurrence in the 8 above types following surgery were 5/11, 0/5, 7/15, 5/8, 0/3, 0/3, 2/3 and 1/4 respectively. Five cases recurred in 21 patients with tumor cells infiltrating into the connective tissue surrounding the tumors. 12 cases recurred in 16 patients with tumor cells infiltrating thru the capsule into surrounding soft tissue. The recurrence rate in patients who had their tumor enucleated or curetted was 56.5% (13/23). The recurrence rate in the tumors resected together with involved bone segment was 24.1% (7/29). Multiple recurrences occurred in 3 cases whose tumors were not well differentiated and had atypical cells. CONCLUSIONS: Recurrence was more often seen in the follicular, acanthoma, basaloid, and keratoid types of ameloblastoma. When these tumors invade adjacent soft tissues, the recurrence rate is also higher. Recurrence is linked to conservative surgery and to tumors with low differentiation. PMID- 11869519 TI - [The effect of defibrase in prevention of experimental atherosclerosis in rabbits]. AB - OBJECTIVE: To study the anti-atherosclerosis effect of difibrase in rabbits. METHODS: Use of computer, light microscope, electron microscope, biochemistry and histochemical methods to identify the total cholesterol (TC) content, mean plaque area in aorta and its cholesterol content, degree of stenosis of coronary arteries of the experimental animals and control animals. RESULTS: Defibrase could decrease the mean plaque area of aorta and the TC content in the experimental animal group was less than that in the control group (P < 0.01). The findings in aortic arches and thoracic aorta were similar to those in the aorta (P < 0.05, P < 0.01). The number of plaques at the small artery openings in the thoracic aorta and abdominal aorta of the experimental group were less than that of the control group (P < 0.01). The cross sections made at the upper third of all hearts showed that the degree of stenosis of coronary artery branches in the heart wall of the experimental group was much less than that of the control group. CONCLUSION: Defibrase can significantly inhibit atherosclerosis. PMID- 11869520 TI - [Immunocyte reaction in stage III nasopharyngeal carcinoma in relation to biological findings and prognosis]. AB - OBJECTIVE: To study the relationship between macrophages (MP), Langerhans cells (LC), T and B lymphocytes, biological behavior and prognosis of stage III nasopharyngeal carcinoma (NPC) patients. METHODS: 43 cases with stage III NPC were studied by immunohistochemical SP staining methods. RESULTS: The positive expression rates of infiltrating LC and/or MP in NPC were 41%, 65% (CD68+), 46% (lysozyme+), and were related to prognosis, but not to age, sex, metastasis to cervical lymph nodes and basi-cranial destruction and/or cranial nerve damage. In addition, infiltrating T or B cells in NPC were not related to the prognosis or biological behavior. However, when both infiltrating T lymphocytes and LC and/or MP were marked, the prognosis of these cases were better than that of those cases whose infiltrating LC and/or MP were marked but T cell infiltration was scarce. CONCLUSION: The prognosis of stage III NPC is related to LC or MP, survival is especially longer in those with marked T cell infiltration. PMID- 11869521 TI - [Clinicopathological study of mucosa-associated lymphoid tissue lymphoma of salivary glands]. AB - OBJECTIVE: To study the histopathological features and pathogenesis of mucosa associated lymphoid tissue lymphoma (MALT-oma) of salivary glands. METHODS: Surgical specimens from 27 cases of MALT-oma of salivary glands were studied using paraffin-embedded section, immunohistochemical method (SP method) and electron microscope. RESULTS: 12 tumors were located in the pariotid glands, the other 15 in the submandibular glands. Much of the MALT-oma of the salivary glands were composed of diffuse centrocyte-like (CCL) cells with "lymphoepithelial lesions". Immunohistochemically, CD20 expression was found to be positive and CD45RO expression was negative in all MALT-omas of salivary glands. CONCLUSIONS: Most of the MALT-omas are low grade malignant tumors and the tumor cells have a "homing back" phenomenon. A small number of MALT-omas can become highly malignant tumors with poor prognosis. Acquired MALT may develop as a reaction to autoimmune disease and/or infection. Hyperimmune reaction and MALT hyperplasia under continuous stimulation may result in myoepithelial sialadenitis and develop into MALT-oma of salivary glands. PMID- 11869522 TI - [A clinico-pathological study of granulomatous amoebic encephalitis]. AB - OBJECTIVE: To study the clinico-pathological characteristics and differential diagnosis of granulomatous meningioencephalitis caused by acanthamoeba. METHODS: The clinical features, CT scan findings and post mortem pathological changes of 3 patients afflicted with granulomatous amoebic encephalitis (GAE) were analysed. RESULTS: GAE is a chronic clinical entity. Clinical manifestations include headache, low fever, seizures, hemiparesis and coma leading to death. CSF: pressure slightly elevated with presence of lymphocytes and plasma cells, glucose usually low with moderately high protein levels. CT scans were non specific. Clinical course insidious and may mimic bacterial meningitis or tuberculous meningitis. Post mortem pathological examinations showed that acanthamoeba produced a chronic or subacute granulomatous meningioencephalitis. Acanthamoebic trophoziotes and cysts were found within the hemorrhagic necrotic cerebral tissues. In some cases there is severe angitis and fibrinoid necrosis of the vascular wall with perivascular cuffing by lymphocytes and plasma cells. Trophozoites and cysts were found within the perivascular space. CONCLUSIONS: Acanthamoeba is an opportunistic pathogenic free living amoeba which can produce fatal GAE. PMID- 11869523 TI - [A clinico-pathological and etiological study of Binswanger's Disease]. AB - OBJECTIVE: To study the pathology, incidence and etiology of Binswanger's disease. METHODS: Autopsied brains from 9 cases of Binswanger's disease and 13 cases of non-demented elderly individuals were studied by clinico-pathological, immunohistochemical and micrometer methods. RESULTS: The lesions of Binswanger's disease were mainly located in the subcortical white matter, periventrical regions, brainstem and cerebellum. The pathological changes were demyelination, lacunar infarction, gliosis and dilatation of perivascular space (Virchow-Robin space, VR space, P < 0.01). The wall thickness of deeply penetrating arteries in the white matter was significantly increased (P < 0.01). CONCLUSION: Binswanger's disease is not a rare entity. The findings of this study suggest that arteriosclerosis is a primary factor in the pathogenesis of Binswanger's disease. PMID- 11869524 TI - [Expression of angiogenic factors and cell cycle regulation factors in human glioblastoma cell line SHG-44]. AB - OBJECTIVE: To investigate the biological features and immunophenotypes of human glioblastoma cell line SHG-44 after long term passage. METHODS: Immunohistochemistry and in situ hybridization were used to study the proliferative activity, intermediate filament protein coexistence, expressions of oncoprotein, angiogenic factors and cell cycle regulation factors. RESULTS: After 130 to 150 passages, SHG-44 cells were weakly positive for glial fibrillary acidic protein (GFAP), but strongly positive for vimentin. The labeling index of Ki-67 and PCNA were 83.5% +/- 10.2% and 70.0% +/- 18.7% respectively. Overexpression of p21 ras, c-erbB-2, epidermal growth factor (EGF) and EGF receptor were obtained. Basic fibroblast growth factor (bFGF), FGF receptor, vascular endothelial growth factor (VEGF) and inducible nitric oxide synthase (iNOS) were also up-regulated in this cell line. p16, p53, cdk4 and cyclin D1 could be detected in the cells and their indices were 43.1% +/- 11.2%, 20.7% +/- 6.6%, 33.1% +/- 11.4% and 29.2% +/- 4.7% respectively. CONCLUSION: Expressive abnormalities of these growth factors, their receptors and the above oncoproteins as well as disorders of cell cycle regulation contribute to the rapid growth and high degree of malignancy of this cell line. PMID- 11869526 TI - [The analysis of pathological changes of cardiac conduction system in 54 cases of cardiac sudden deaths]. AB - OBJECTIVE: To elucidate the exact cause of death in cases of cardiac sudden deaths. METHODS: Sampling of cardiac conduction system (CCS) designed by the research group was adopted. The CCS of 120 cases died of cardiac sudden deaths were collected for routine histological examination. RESULTS: In 54 out of 120 cases, the causes of death were attributed to lesions present in the CCS, and 85.2% of them were young adults. The peak age range was from 20 - 30. Much more in males (6.7:1). The pathological changes in CCS include: inflammation (46.2%), fatty infiltration, hypertrophic cardiomyopathy associated with CCS changes; hemorrhage, developmental anomalies, fibrosis and tumors. CONCLUSION: The pathological changes of CCS are relatively common, as the cause of sudden deaths. Which is considered to be the pathological basis of the unstable cardiac electric potentials and arrhythmia. This study recommended the necessity of routine examination on CCS in cases of sudden deaths. PMID- 11869527 TI - [Expression of membrane-type I matrix metalloproteinase and its correlation with gelatinase A activation in cancer cell lines]. AB - OBJECTIVE: To study the expression of membrane-type I matrix metalloproteinase (MT1-MMP) and its correlation with gelatinase A (MMP2) activation. METHODS: Reverse transcription PCR, northern blots and western blots were employed to detect the expression levels of MT1-MMP mRNA and MMP2 protein in 4 human melanoma cell lines, 2 human lung cancer cell lines and 2 human prostate cancer cell lines. RESULTS: All the cancer cell lines expressed MT1-MMP mRNA, and the MT1-MMP mRNA level in the 4 melanoma cell lines was significantly higher than that in the lung and prostate cancer cell lines. Activated MMP2 proteins were only detected in the melanoma cell lines, whereas PG, a lung cancer cell line, which secreted pro-MMP2 and expressed low level of MT1-MMP, could not produce activated MMP2. CONCLUSIONS: The level of MT1-MMP expression was highly associated with MMP2 activation. PMID- 11869525 TI - [Observations on expression of c-fos and c-myc genes and activity of PDGFBB autocrine loop in 67 human gliomas]. AB - OBJECTIVE: To investigate the variance of expression of c-fos and c-myc genes and the activity of platelet-derived growth factor (PDGF) BB autocrine loop in human gliomas as well as their relationship. METHODS: In situ hybridization and immunohistochemistry were used to study 67 human glioma specimens of different grades. RESULTS: The incidences, of positive expression for c-fos mRNA, c-fos protein, c-myc mRNA and c-myc protein were 100% (67/67), 100% (67/67), 85.1% (57/67) and 83.6% (56/67) respectively. There was significant positive correlation between the positive cell densities of c-fos mRNA (92.49 +/- 33.87) and c-fos protein (91.64 +/- 38.11), and of c-myc mRNA (72.70 +/- 40.56) and c myc protein (71.49 +/- 41.11, r = 0.917 - 0.988, P < 0.001). All of them also increased correspondingly with the malignant degree of the tumors and level of PDGFB mRNA expression in the tumor cells (P < 0.05 - 0.01). Not only were the positive cell densities of PDGF alpha receptor (58.46 +/- 28.77), PDGF beta receptor (63.03 +/- 31.63) and phosphotyrosine protein (68.25 +/- 29.30) correlated positively with one another (r = 0.827 - 0.896, P < 0.001), but they also all correlated positively with the positive cell densities of PDGFB mRNA, c fos mRNA, c-fos protein, c-myc mRNA and c-myc protein respectively (r = 0.615 - 0.888, P < 0.001). CONCLUSIONS: The abnormally increased activity of the PDGFBB autocrine loop possibly upregulated the overexpression of c-fos and c-myc genes in glioma cells and suggested that c-fos and c-myc proteins probably played an important role as downstream response factors in the signaling pathway in which PDGFBB could induce gliocyte transformation and glioma development. PMID- 11869528 TI - [A clinico-pathological and immunohistochemical study on anaplastic large cell lymphoma]. AB - OBJECTIVE: To study the clinicopathological characteristics, immunophenotype and biologic behavior of anaplastic large cell lymphoma (ALCL). METHODS: Clinicopathological and immunohistochemical methods (ABC method) were used to analyse 8 cases of ALCL and followed for 6 to 25 months. RESULTS: Eight ALCL patients (4 male and 4 female) accounted for 4.1% of the 195 lymphomas seen from 1994 to 1997. Average age was 40 years. Two tumors were located in the skin, the remainder were systemic. Clinical stage: stage I, 2 cases; stage II, 1 case; stage III, 5 cases. Histopathology: Tumor cells exhibited high pleomorphism and a large amount of mitotic figures. Reactive components (cells) often observed in the background. Immunohistochemical studies found that all tumors reacted positively with CD30 and negatively with CD15. Five cases expressed T-cell's markers (CD3, UCHL-1, CD43) and three expressed B-cell markers (CD79a, L26). After chemotherapy, two patients with cutaneous type ALCL had complete remission and survived 14 and 25 months respectively. Of the six systemic cases, three survived for 6-8 months, the remaining three died at 2 days, 20 days and 18 months after diagnosis. CONCLUSIONS: This study found that ALCL was a group of lymphomas showing similarity in morphology but diversity in clinical presentation and immunophenotype. Even if the tumor is very responsive to chemotherapy, ALCL remains a high grade malignant lymphoma. PMID- 11869529 TI - [A preliminary report on two distinct tumor-suppressor regions on chromosome 1p36.2-p36.3 in human hepatocellular carcinomas]. AB - OBJECTIVE: Both cytogenetic and molecular genetic analyses have unveiled non random genomic alterations in the distal short arm of human chromosome 1 associated with a number of human malignancies including heptatocellular carcinoma (HCC). The aim of this investigation is to determine the precise region of deletion that may harbor the putative tumor suppressor genes in HCC. METHODS: For the study of the loss of heterozygosity (LOH), 38 cases of hepatitis B virus (HBV) associated HCC and their corresponding non-tumor liver tissues were detected with 43 microsatellite polymorphic markers particularly focusing on 1p. RESULTS: Twenty-eight of the 38 (74%) tumors showed LOH on at least one locus on 1p36.2-p36.3. Two distinct smallest common deleted regions (SCDRs) with different patterns of deletion were identified. The first SCDR is located on locus D1S2795 at 1p36.3, between loci D1S2145 and D1S2893. The second SCDR is located at 1p36.2, between loci D1S244 and D1S489. Both of the SCDRs have not been previously described in HCC. In addition, a region of possible homozygous deletion (HD) was also detected within the second SCDR between loci D1S1597 and D1S489 by comparative multiplex PCR. This is the first observation of a possible homozygous deletion on the distal short arm of chromosome 1 in HCC as well as in human tumors. CONCLUSIONS: The high-resolution deletion mapping of 1p36.2-p36.3 in HCC in this study confirmed the presence of two distinct regions of deletion. Our data strongly suggest the presence of at least two tumor suppressor regions on 1p36.2-p36.3 and play an important role in the pathogenesis of HBV associated HCC. These results also provide a basis for further studies directed at cloning potential tumor suppressor genes in these regions. PMID- 11869530 TI - [The effects of hypoxia on vascular pericytes of the lung]. AB - OBJECTIVE: To study the effects of hypoxia, directly or mediated by pulmonary arterial endothelial cells, on energy metabolism, cell cycle, platelet derived growth factor (PDGF) and PDGF receptor mRNA expression of vascular pericytes of the lung in vitro. METHODS: Use of MTT colorimetric assay, flow cytometry, nucleic acid in situ hybridization and automatic image analysis for quantitative analysis. RESULTS: Hypoxia promoted vascular pericyte proliferation directly or through the mediation of endothelial cells to promote vascular pericyte proliferation from static phase (G(0), G(1) phase) to DNA synthesis phase (S phase) and mitotic phase (G(2) + M phase) and upregulate gene expression of PDGF and PDGF-R by vascular pericyte of the lung. CONCLUSIONS: The low oxygen tension of alveolar gas and hypoxemia may promote transcription of PDGF and its receptor gene. Paracrine and autocrine PDGF may induce proliferation of vascular pericytes of the pulmonary vessels and may play a role in pulmonary hypertension and structural remodeling of the pulmonary vessels, especially in the neomuscularization of nonmuscular pulmonary arterioles. PMID- 11869531 TI - [A pathological study on the correlation of HPV infection with precancerous cervical lesions]. AB - OBJECTIVE: To study the correlation of cervical condyloma and cancerous lesions with HPV infection. METHODS: Cervical biopsies and histopathological examinations were performed on 179 cases which had abnormal cervical cytological smears. PCR was used to study the HPV-DNA of 128 cases and in situ hybridization (ISH) was used to study 10 cases. RESULTS: 1. Morphologic observations. Most cervical condyloma cases were of the morphologically flat type (97.7%). Two koilocyte types were observed, the classical, or so called diagnostic koilocyte type (39.7%) and the atypical type (60.3%). Cervical condyloma often occurred together with cervical intraepithelial neoplasia (CIN, 42.5%). 2. PCR HPV DNA subtype analysis. Of the 58 cervical flat condyloma, 5 were PHV6/11 positive (8.6%) and 28 were HPV16/18 positive (48.3%). 86.1% of those with atypical koilocyte and 9.1% of those with diagnostic koilocyte had HPV16/18 infection. 66.7% of the lesions in which condyloma coexisted with CIN(2-3) had HPV16/18 infection. CONCLUSIONS: Most cervical condyloma lesions were of the flat type. The appearance of atypical koilocyte is correlated to HPV16/18 infection, which in turn is correlated to the degree of CIN malignancy. PMID- 11869532 TI - [Clinical pathological analysis and immunohistochemical expression of primary cervical adenocarcinoma in 98 cases]. AB - OBJECTIVE: To discuss the pathological classification, immunohistochemical expression characteristics, differential diagnosis and factors affecting prognosis in cervical adenocarcinoma. METHODS: Clinical pathological analysis and immunohistochemical study were performed on all primary cervical adenocarcinoma cases (98 patient) from 1991 to 1998. Comparison was made with 18 cases of endometrial adenocarcinoma. 54 cases were followed-up. RESULTS: These primary cervical adenocarcinoma cases fell into 5 categories, of which the endocervical type being the most common. The positive rate of CEA, HPV16 and HPV6B/11 staining were 15/18 (83.3%), 6/9 (66.7%) and 10/18 (55.6%) cases respectively, while the results of ER and vimentin staining were all negative. PCNA staining was strongly positive in all the cases. In the control group (endometrial adenocarcinoma group) the CEA and vimentin staining positive rates were 27.8% and 83.3% respectively, the difference was significant. The two year follow-up indicated that the incidence of recurrence, metastasis and mortality in the endocervical type were higher that those of other types. CONCLUSION: It is difficult to diagnose highly differentiated endocervical adenocarcinoma preoperatively, therefore adequate attention should be attached. Immunohistochemical staining can be used as an important basis for the diagnosis and differential diagnosis of cervical adenocarcinoma. The strongly positive results of PCNA staining, regardless of the degree of differentiation, proves that even highly differentiated adenocarcinoma exhibits a high degree of cell activity. Preliminary data suggests that cervical adenocarcinoma is related to HPV infection. Its relation with each HPV subtype remains to be clarified. PMID- 11869533 TI - [Morphometric study of inflammatory cells and smooth muscle cells in coronary plaques and their relation with plaque stability]. AB - OBJECTIVE: To compare the content of inflammatory cells and smooth muscle cells in stable and unstable coronary plaques in order to clarify the role of these cells in the vulnerability of plaques. METHODS: Hearts from 12 postmortem acute myocardial infarction cases were studied. The epicardial coronary arteries were dissected en bloc, fixed, decalcified, cut at 4 mm intervals and routinely processed for HE sections. 163 stable plaques (with no or only little lipid core) and 163 unstable plaques (with lipid core size > 40% of plaque area) were studied immunohistochemically using monoclonal antibodies specific for macrophages (CD68), T lymphocytes (UCHL-1) and smooth muscle cells (actin). Computor aided planimetry was used to measure the positive area of different cells. RESULTS: The content of macrophage and T lymphocytes in unstable plaques were significantly higher than that in stable plaques (P < 0.05). CONCLUSIONS: Stable plaques and unstable plaques not only had different lipid core size but also had different inflammatory cell and smooth muscle cell content. The larger the lipid core and the more macrophages and T lymphocytes in the fibrous cap, the more unstable the plaque. The lipid core and inflammatory cells (including macrophages and T lymphocytes) are the two major determinants of the vulnerability of coronary plaques. PMID- 11869535 TI - [A study on the loss of heterozygosity of multiple tumor suppressor genes in gastrointestinal metaplasia]. AB - OBJECTIVE: To investigate the role of multiple tumor suppressor gene abnormality in gastrointestinal metaplasia (IM). METHODS: The loss of heterozygosity (LOH) of YNZ22, APC, MCC and DCC genes was examined by PCR-RFLP technique in 60 IM cases. RESULTS: LOH of YNZ22, APC, MCC and DCC genes were found in 19.4% (6/31), 12.0% (3/25), 13.0% (3/23) and 4.3% (2/47) respectively. IM was classified into types I, II and III. It was found that the LOH rates of each tumor suppressor gene of type III IM were higher than those of types I & II IM, but the difference was not significant (P > 0.05). Two cases of type III IM were found to also have 2 gene alterations (APC, MCC or YNZ22, MCC) respectively. All LOH were found in IM of adjacent carcinoma. None were found in IM of cancer-free patients. CONCLUSION: The alteration of tumor suppressor gene may play a role in the development and carcinogenesis of IM. PMID- 11869536 TI - [Apoptosis of HL60 cells induced by inhibition of bcl-2 with antisense phosphorothioate oligodeoxynucleotides]. AB - OBJECTIVE: To observe the effects of different bcl-2 antisense phosphorothioate oligodeoxynucleotide (ASPO) concentrations in inducing apoptosis of HL60 cells. METHODS: Signs of apoptotic cells were detected by acridine orange staining, transmission electron microscope, flow cytometry and DNA fragment electrophoresis. RESULTS: In presence of ASPO, the apoptosis of HL60 cells could be induced after 24 hours of incubation. The apoptosis rate at ASPO 5, 10 and 20 micromol/L were (9.67 +/- 2.16)%, (18.25 +/- 3.01)% and (23.42 +/- 2.75)% respectively. There was a statistically significant increase in apoptosis as the dose of ASPO increased. After incubation for 72 hours, the apoptotic rates of the 3 different ASPO concentration groups were (9.58 +/- 2.96)%, (29.58 +/- 4.04)% and (36.33 +/- 3.66)% respectively, of which the apoptic rate of HL60 cells incubated with ASPO 5 micromol/L was highest at 48 hours. CONCLUSION: Bcl-2 ASPO can specifically induce apoptosis of HL60 cells. Its effect depends upon the concentration of ASPO. PMID- 11869534 TI - [mRNA and protein expression of apoptosis-regulating gene bcl-x in lymphomas]. AB - OBJECTIVE: To observe mRNA and protein expression of bcl-x and explore the role of bcl-x in the regulation of apoptosis and oncogenesis and their value in differential diagnosis of lymphomas. METHODS: Reverse transcription-polymerase chain reaction (RT-PCR) and citric acid-microwave-ABC immunohistochemistry (ICH) were used to observe mRNA transcription forms and protein distributions of bcl-x in 30 frozen tissues and 109 paraffin embedded tissues (both including reactive hyperplasia and common types of lymphoma). RESULTS: RT-PCR revealed that all 30 frozen cases showed bcl-x transcription, of which 29 had bcl-x1 band and 13 had bcl-xs band. Bcl-x1 band was always stronger than bcl-xs band. IHC found that 97/109 cases had fine positive bcl-x granules in the cytoplasm. In lymphoid reactive hyperplasia, bcl-x exhibited weak immunoactivity in cells located in the germinal centers and interfollicular regions of the nodes but not in the mantle zone lymphocytes which surround germinal centers. In lymphomas, bcl-x immunoactivity was always strong and diffuse. The staining fractions in different types of lymphomas were significantly higher than those in lymphoid reactive hyperplasias (P > 0.05), but no significant difference was found between the positive rate and staining fraction of different types of lymphomas. CONCLUSION: Bcl-x shows active mRNA transcription and high protein expression in lymphomas. The long isoform of bcl-x (bcl-xl) is the predominant expression. The characteristics of bcl-x protein expression may be helpful in differential diagnosis of reactive hyperplasia and lymphoma. PMID- 11869537 TI - [Effects of p21WAF1/CIP1 gene introduction on a human lung cancer cell line with p53 mutation]. AB - OBJECTIVE: To study the effects of p21WAF1/CIP1 gene expression on the biological hehavior of cancer cells with p53 gene mutation. METHODS: Expression of p21 vehicle was constructed and introduced into a human lung cancer cell ling PG with p53 gene mutation, the morphological and growth characteristics of the transfectant were then analysed. The CDK4 and PCNA levels were evaluated and the response to genotoxic agents were observed. RESULTS: The PG cells with mutant p53 were transfected with ectopic p21 gene which resulted in overexpression of p21 (6 fold of the control). The transfectants presented decreased nuclear irregularity and thinner membrane in nuclear atypia, with delayed growth rate and contact growth inhibition. The transfectant also displayed high serum requirement in culture and increased anchorage dependence. Both levels of CDK4 and PCNA were decreased. However, the cisplatin induced cell apoptosis was delayed. CONCLUSIONS: Expression of p21WAF1/CIP1 gene was effective in cancer suppression even in p53 gene mutation. This may have been achieved through lowering of CDK4 or PCNA levels and not involving the apoptosis mechanism. Activation of p21WAF1/CIP1 gene may be considered a bypass for restoration of p53 gene function in cancer cells with p53 gene mutation. PMID- 11869539 TI - [Protective effect of endothelin receptor subtype A antagonist BQ610 on the ultrastructure of acute ischemic myocardium in cats]. AB - OBJECTIVE: To observe the effects of endothelin receptor subtype A antagonist BQ610 on acute ischemic myocardial injury by electronic microscopy to study whether endogenous endothelin is involved in the pathogenesis of acute myocardial ischemia. METHODS: Acute myocardial ischemia was induced by ligation of the left anterior descending coronary artery (LAD) in 12 cats, and BQ610 was immediately injected into the occluded LAD. Controls received normal saline (NS) injection. The ultrastructure of ischemic myocardium and non-ischemic myocardium were observed 30 min after LAD ligation. RESULTS: As compared wtih NS controls, BQ610 significantly protected the acute ischemic myocardium from ultrastructural injury, including cardiac muscle fiber swelling, myofibril disorganization and breakdown, mitochondrial swelling and endothelial swelling. In general, the non ischemic myocardial ultrastructure was normal. CONCLUSIONS: The results indicate that endogenous endothelin, through endothelin receptor subtype A, plays an important role in ultrastructure injury of acute ischemic myocardium in cats. PMID- 11869538 TI - [Transformation of human bronchial epithelium by EB virus gene LMP1 and EBNA2]. AB - OBJECTIVE: To observe Epstein-Barr virus (EBV)-dependent transformation of human bronchial epithelium. METHODS: Eukaryotic expression plasmide pBS-LMP-Hyg and pSG5-EBNA2-Hyg were transfected into immortalized human bronchial epithelium cell line TR. RESULTS: In situ hybridization and Western blot confirmed EBNA2 and LMP1 expression in one hygromycine B resistant cell clone TR/LMP1-EBNA2. Growth curve and MTT value showed that transformed cells grew faster. Soft agar plating rate of TR/LMP1-EBNA2 was 50.1per thousand, 68.2per thousand, empty vector PBs-Hyg transfected cell strain TR/Hyg 2.0per thousand, 3.0per thousand, TR 11.0per thousand, 0.7per thousand. CONCLUSION: Expression of both LMP1 and EBNA2 led to decreased anchorage dependence and increased proliferation rate of TR, thereby further transformed immortalized human bronchial epithelium. Our results suggest that EBV may play a role in genesis of lung cancer. PMID- 11869540 TI - [Effects of EB virus LMP on the differentiation and growth of human nasopharyngeal carcinoma cell line CNE1]. AB - OBJECTIVE: To study the effects of EBV-LMP gene on differentiation and growth of human nasopharyngeal carcinoma (NPC) cell. METHODS: With NPC cell line (CNE1) as target, electroporation was used to transfect EBV-LMP gene and the vector into CNE1 cells. The proliferation changes of transfectants were measured in vitro by proliferation experiment. FCM method and transplantation in nude mice were used. RESULTS: LMP promoted the growth ability of NPC CNE1 cells in vitro. The average A ratio (3.98 +/- 0.11) in the treatment group was higher than in the control group (2.36 +/- 0.05) and also in the negative control group (2.75 +/- 0.07, P < 0.01). Clone forming test (25.2%, 378/1 500) in the treatment group was higher than in the control groups (11.2%, 168/1 500) and negative control groups (13.4%, 201/1 500). When FCM method was used, the keratin positive rate (82.7%) in the treatment group was markedly decreased as compared with the control (92.5%) and negative control groups (95.7%, P < 0. 05). Latent period (26.6 +/- 7.7) days and internal double time (2.51 +/- 0.18) days of transplant tumor of the treatment group were markedly shortened, tumor take rate (5/6) was increased, cells of the transplant tumor in the treatment group showed an image of small size, great allotype, and a trend of low differentiation. CONCLUSION: The EBV-LMP can promote the growth and inhibit the differentiation of NPC CNE1 cells. EBV-LMP may play an important role in the development of NPC and in studying its mechanism. PMID- 11869541 TI - [Mucocele-like lesion of the breast-histopathology and fine needle aspiration cytology changes]. AB - OBJECTIVE: To elucidate the histopathology and fine needle aspiration cytology (FNAC) features of mucocele-like lesion (MLL) of the breast, and to establish the criteria in separating MLL from mucinous carcinoma on the basis of pre-operative FNAC. METHODS: Seven cases of MLL (5 benign and 2 malignant) were reviewed. The histology and FNAC features were compared with 14 cases of mucinous carcinoma examined in the same period. RESULTS: All cases of MLL presented with breast lumps (1 case was detected by doctor after abnormal mammogram) and MLL patients are much younger (mean age 34.8 years for MLL and 67.9 years for mucinous carcinoma). Histologically, MLL are characterized by mucinous cysts which may rupture and discharge their contents into the surrounding tissue. The lining epithelium is bland and epithelial cells are rare in the mucin pool. Malignant MLL shows associated in-situ carcinoma. In contrast, mucinous carcinoma is formed by large mucin pools devoid of lining epithelium and contains mumerous tumor cells in the mucin pools. In all cases, grossly visible jelly-like mucous was obtained from FNA. Cytologically, all smears showed abundant mucous in the background. The most important fratures distinguishing benign MLL from mucinous carcinoma were scant cellularity, rare intact single tumor cell and tumor cells arranged in cohesive monolayered clusters lacking significant nuclear atypia. CONCLUSION: Mucinous lesions of the breast can be divided into MLL and mucinous carcinoma based on the FNAC changes. Operative therapy can be performed for definite cases of mucinous carcinoma without further biopsy. However, excisional biopsy is advised in MLL for further separation into benign and malignant categories. PMID- 11869542 TI - [Histological grading in ductal carcinoma in situ of the breast]. AB - OBJECTIVE: To study the significance of histological grading as a prognostic factor in ductal carcinoma in situ of the breast. METHODS: According to Van Nuy's classification, 32 cases of ductal carcinoma in situ (DCIS) of the breast were divided into three grades. RESULTS: Low grade (well differentiated, low grade DCIS) 12 patients (37.5%); Intermediate grade, 9 patients (28.1%); High grade (poorly differentiated DCIS) 11 patients (34.4%). Among the high grade DCIS, the histological subtypes were comedo (9 patients), micto papillary (1 patient) and solid (1 patient). The positive expression of c-erbB-2, p53 and MIB-1 in high grade DCIS was higher than that in intermediate and low grade DCIS. The difference between high grade and low grade DCIS was significant (P < 0.05). The expression of ER in high grade DCIS was lower than that in intermediate and low grade DCIS. CONCLUSIONS: Histological grading of breast ductal carcinoma in situ may be a good prognostic factor. PMID- 11869543 TI - [Significance of telomerase activity detection by fine-needle aspiration in patients with breast cancer]. AB - OBJECTIVE: To study the telomerase activity in samples of breast fine-needle aspiration specimens and to investigate its potential clinical implications. METHODS: Ninety nine fine-needle aspiration specimens from 83 patients with breast cancers, 12 with benign lesions and 4 with breast inflammation, were studied for the presence of telomerase activity by a PCR-based telomere repeat amplification protocol (PCR-TRAP). RESULTS: Among 83 fine-needle aspiration specimens from patients with breast cancer, telomerase activity was detected in 61 of 69 cytologic-positive specimens, 5 of 7 cytologic-suspicious specimens and 4 of 7 cytologic-negative specimens. Sensitivity of telomerase activity detection was 84.3% (70/83); the coincidence rate between telomerase detection and cytologic confirmation was 77.1% (64/83). Of 12 specimens from patients with benign breast lesions, 4 were positive for telomerase activity, but 4 specimens from patients with breast inflammation were all negative. The Combined use of cytological and telomerase detection can improve the detection rates from 83.1% (69/83) to 93.9% (78/83). CONCLUSION: Detection of telomerase in breast tissue samples by fine-needle aspiration is a sensitive and specific method, which implies that it would be helpful in preoperative cytological diagnosis of breast cancer. PMID- 11869544 TI - [A clinicopathological study of pancreatoblastoma]. AB - OBJECTIVE: To investigate the clinicopathological, immunohistochemical features and the histogenesis of pancreatoblastoma (PB) in 14 pediatric cases, including 9 male and 5 female patients, their age ranged from 1.5 to 8 years with a mean of 4.4 years. METHODS: Routine pathological, histochemical, and immunohistochemical methods were utilized to analyse the PB specimens. RESULTS: Tumors of 6 patients had metastasized to the liver, spleen and lymph nodes, 8 patients were alive 10 months to 105 months after diagnosis. Histologically, the tumors were composed of dense epithelial elements separated by fibro-stromal tissue, resulting in a nesting or organoid pattern. Immunohistochemically and histochemically, the tumors exhibited acinar, endocrine and ductal differentiation. The positive rates for were: CK 100%, EMA 86%, NSE 79%, CEA 71%, SYN 64%, CgA 43%, alpha1-AT 57%, PAS 100% and mucicarmine 57%. CONCLUSIONS: PB is the most common malignant pancreatic neoplasm of childhood. PB arises from primitive pluripotential cells capable of differentiating into 3 pancreatic cell types: acinar, endocrine and ductal. The prognosis is better than pancreatic carcinoma in adults. PMID- 11869545 TI - [Effects of LMP-1 expression on neoplastic cell proliferation and apoptosis in nasopharyngeal carcinoma]. AB - OBJECTIVE: To investigate the effects of LMP-1 expression on neoplastic cell proliferation and apoptosis in untreated nasopharyngeal carcinoma (NPC). METHODS: Biopsies from 55 untreated NPC cases were collected. EBERs was detected by in situ hybridization. PCNA, LMP-1 and bcl-2 were detected by immunohistochemistry method. Cell apoptosis was demonstrated by TUNEL (TdT-mediated dUTP-x nick end labeling) kit. RESULTS: (1) The positive LMP-(1) expression rate was 27/55 (49.1%). The median PI (PCNA Index) of LMP-1 positive cases (90.0%) was higher than that of LMP-1 negative cases (80.0%) and the p value was less than 0.05 by rank sum (rs) test. (2) The median TI (TUNEL Index) of LMP-1 positive cases (16.2 per high power field) was also higher than that of LMP-1 negative cases (10.4 per high power field) and the p value was less than 0.05 by rs test. (3) As for the bcl-2 expression percentages, no correlation was found between LMP-1 positive and negative cases. (4) A positive rank correlation existed between PI and TI (rs = 0.368 1, P < 0.01), but no correlation was found between TI and bcl-2 expression percentage. CONCLUSIONS: (1) NPC with LMP-1 expression have higher levels of PI, maybe resulting from the promotional effect of LMP-1 on cell proliferation. (2) NPC with LMP-1 expression also have a higher level of TI, indicating that LMP-1 can induce cell apoptosis, which could not be blocked by bcl-2 expression. The authors suggest that LMP-1 expression may exert a dual action on neoplastic cell proliferation and apoptosis in untreated nasopharyngeal carcinoma. PMID- 11869546 TI - [The significance of endometrial biopsy during luteal median phase in diagnosis of luteal function]. AB - OBJECTIVE: To investigate the significance of progesterone profile and endometrial biopsy during luteal median phase in diagnosis of luteal function. METHODS: The progesterone profile during median luteal phase and endometrial histology during median and late luteal phase of 53 infertile women with infertile and recurrent abortion for unknown reasons were studied. RESULTS: Retarded endometrial development occurred in the median luteal phase in 24 of the 53 cases, 9 of which lasted into the late phase. The average progesterone profile of these patients was markedly lower compared with the control group (P < 0.05). However, the average progesterone profile remained normal in the other 13 cases, in whom, RED got normalized in the late luteal phase. Endometrial biopsies taken during the median and late luteal phase from the rest of 29 cases were normal. CONCLUSIONS: In patients with retarded endometrial development during the full luteal phase, luteal function was decreased, but in patients with retarded endometrial development only during the median luteal phase, luteal function appeared to be normal. PMID- 11869547 TI - [Epstein-Barr virus infection and expression of T-cell intracellular antigen-1 (TIA-1) in intestinal T-cell lymphoma]. AB - OBJECTIVE: To study the status of Epstein-Barr virus (EBV) infection and the expression of TIA-1 antigen in 24 cases of intestinal T-cell lymphomas. METHODS: In situ hybridization for EBER1/2; immunohistochemical staining by three step ABC technique for TIA-1, LMP-1, CD30, CD45RO, CD3 AND CD20. 19 cases of intestinal B cell lymphomas were used for comparison. RESULTS: (1) Of the 24 intestinal T-cell lymphoma cases, 23 were pleomorphic, medium to large cell type, and one cases was small cell type; in 20 of the 24 cases EBER1/2 signals were detected in the tumor cells; in 4 of the 20 EBER1/2 positive cases, tumor cells expressed LMP-1; EBER1/2 was not detected in 19 cases of intestinal B-cell lymphomas. (2) in 23 of the 24 cases, TIA-1 was expressed in most tumor cells, but not in the only case of small cell type; TIA-1 was negative in tumor cells of all 19 cases of intestinal B-cell lymphomas. CONCLUSIONS: A high level of EBV infection was present in this group of intestinal T-cell lymphomas, the expression of TIA-1 may be related to its cell origin. Intestinal T-cell lymphoma may be an EBV associated extranodal NK/T cell lymphoma. PMID- 11869548 TI - [Cloning whole length cDNA of related genes responsible for smooth muscle cells proliferation in atherogenesis and study on its function]. AB - OBJECTIVE: To clone whole length cDNA of the related genes responsible for vascular smooth muscle cell (SMC) proliferation in atherogenesis, and to study its function. METHODS: ox-LDL was added as a stimulant to the SMC culture medium. Subtractive library was established using subtractive hybridization technique in order to clone the related genes fragments. With the whole length cDNA library established, the whole length cDNA of the related gene was cloned. The protein expressed was studied. RESULTS: 4 new gene fragments and one whole length cDNA were cloned. The new cloned gene is able to express a protein of about 44000 daltons and closely related to the activity of ox-LDL. CONCLUSIONS: The new cloned gene is considered responsible for SMC proliferation. PMID- 11869549 TI - [Expression of antisense telomerase genes suppressing human cancer malignant phenotypes]. AB - OBJECTIVE: Altering the expression of telomerase genes hTRT and hTR to induce changes in cancer cell biology and to determine their value in cancer gene therapy. METHODS: By genetic transfection of antisense hTRT into HeLa cells, and treatment of PG cells with antisenes hTR oligonucleotides, their effects on cancer cell growth and malignant phenotypes were analyzed. RESULTS: The vehicle for eukaryotic expression of hTRT antisense was constructed and transfected into HeLa cells. The obtained transfectants T4, T5 that could produce antisense hTRT stablely showed marked decrease in growth, with an arrest rate of 24%; the presence of contact growth inhibition was obvious; in nude mice transplantation, the rate for tumor induction was decreased from 100% to 25% or 0%; histologically, the tumor cells from inoculation of transfectant showed less nuclear chromatins and fewer giant tumor cells than those of the parent HeLa cells;the expression of PCNA was significantly reduced in the transfected cells. Treatment of antisense hTR oligonucleotides inhibited PG cell growth at a rate of 8%. CONCLUSIONS: Antisense expression of telomerase genes could significantly suppress cancer cell growth, decrease malignant phenotypes and histological atypia. Therefore, altering expression of telomerase genes may be a new pathway for cancer therapy. PMID- 11869550 TI - [Isolation and characterization of human prostate cancer cell subclones with different metastatic potential]. AB - OBJECTIVE: To isolate and characterize cancer cell subclones with different metastatic potential from human metastatic prostate cancer cell line (PC-3M). METHODS: Using limited dilution, in vitro growth, Matrigel invasion assay, soft agar cloning and in vivo tumorigenicity and spontaneous metastasis assay in nude mice, were isolated and characterized four subclones with different metastatic potential. RESULTS: Four subclones derived from PC-3M were 1E8, 2E7, 2B6, 2B4. Each subclone exhibited a different metastatic potential when inoculated into nude mice. Among these subclones, 1E8 expressed as the highly metastatic phenotype, with 100% metastasis frequency 5 weeks after subcutaneous inoculation into nude mice, whereas 2B4 was not metastatic. In vitro 1E8 was found to be the most highly invasive cell line in Matrigel invasive assay (98 +/- 24) and had the most clones in soft agar cloning assay (265 +/- 39) while 2B4 was found to have the least invasive abilities (12 +/- 4) and the least clones (137 +/- 14). CONCLUSION: Successful establishment of these subclones with different metastatic potential may be valuable for further study on the molecular mechanisms of cancer metastasis and cloning of cancer metastasis-related genes. PMID- 11869551 TI - [Pathomorphological and amyloid beta-protein immunohistochemical findings in autopsied brains of Alzheimer's disease]. AB - OBJECTIVE: To study the incidence and neuropathological features of Alzheimer's disease in Chinese and the significance of silver and amyloid beta-protein staining for the pathological diagnosis of Alzheimer's disease. METHODS: Among 445 autopsy cases above 60 years in age (including 60) collected from 1982-1998 in the Department of Pathology, Beijing Hospital, totally 29 cases were diagnosed as Alzheimer's disease pathologically (6.5%), accompanying with a history of mental deterioration and the histological changes in certain areas of the brain fulfilling the Khachaturian or Mirra criteria. Tissue blocks were taken from 6 standard regions of the brains for Bodian, Bielschowsky and amyloid beta-protein immunostaining. Senile plaques and neurofibrillary tangles were quantified. RESULTS: Four main types of senile plaques were noticed and the result by amyloid beta-protein immunostaining was considered more sensitive than that of the silver impregnation staining. The former one enabled to demonstrate all the 4 types of senile plaques, while silver impregnation staining could only demonstrate clearly the primitive and neuritic plaques, but not the small diffuse and burn-out plaques. The senile plaques were noticed to be focused in the hippocampus, superior temporal gyrus and middle frontal gyrus while the neurofibrillary tangles, were mainly obtained in the hippocampus, temporal cortex and amygdaloid. Cerebral amyloid angiopathy was found in 25 out of 29 Alzheimer's disease cases. CONCLUSIONS: Alzheimer's disease may be the main cause of dementia among the elders in Chinese and immunostaining for amyloid beta-protein pretreated with formic acid may set a high value on the pathological diagnosis of Alzheimer's disease. PMID- 11869553 TI - [The significance of p16, cyclin D1 and pRb expression in soft tissue leiomyosarcoma]. AB - OBJECTIVE: To evaluate the significance of p16, cyclin D1 and pRb expression in human soft tissue leiomyosarcoma (LMS) and the relationship among these three proteins. METHODS: 42 cases of LMS were studied for p16, cyclin D1 and pRb expression with immunohistochemical method. RESULTS: The total abnormality percentage of p16, cyclin D1 and pRb expression in all 42 cases of LMS was 97.6%. The negative rate of p16 and pRb protein expression was 35.7% and 47.6% respectively, and the overexpression rate of cyclin D1 was 57.1%. Overexpression of cyclin D1 was also noticed in 55% leiomyomas. In the 20 cases of LMS which were pRb negative, 19 cases were moderately or strongly positive and one faintly positive for p16. Contrastly, in the 15 cases of LMS negative to p16, 12 cases were moderately or strongly positive and 3 cases faintly positive for pRb. Higher rates of loss of pRb protein were observed in the low-differentiated group than in the high-differentiated one. CONCLUSIONS: (1) The abnormality of p16-cyclin pRb pathway is highly related to the pathogenesis of LMS. (2) Loss of p16 or pRb and/or overexpression of cyclin D1 may be involved in the pathogenesis of LMS. (3) There is a negative reciprocity between p16 and pRb protein expression in LMS. (4) Overexpression of cyclin D1 is a common molecular alteration that may occur in the early stage. (5) Expression of pRb is inversely related to the differentiation degree of LMS. (6) The expression of the 3 proteins studied has no relation to tumor location, recurrence, invasion or metastasis. PMID- 11869554 TI - [Microsatellite alterations on chromosome 9p21-22 in sporadic colorectal cancer]. AB - OBJECTIVE: To study whether alteration of p16 plays an important role in the development of colorectal carcinomas and the relationship between the molecular changes of 9p21-22 chromosome subregion in sporadic colorectal cancers. METHODS: To detect microsatellite instability (MSI) and loss of heterozygosity (LOH) by PCR, denatured-polyacrylamide gel-electrophoresis and silver staining (microsatellite DNA-PCR-silver staining method) and to compare the results with the clinopathological parameters. RESULTS: Between MSI positive and negative cases and the clinopathological findings, some evidences found in the MSI positive group were as follows: (1) tendency towards younger patients (usually < 50 years in age, P < 0.05); (2) more frequently seen in mucoid carcinomas (P < 0.01). CONCLUSIONS: Microsatellite DNA-PCR-silver staining method is very sensitive in detecting even a tiny change of a single base. MSI occured in the selected microsatellite loci of different subregions and different chromosomes might be different in significance, therefore, a right choice of the suitable loci for studying the microsatellite changes is important. Since the frequency of loss of heterozygosity at 9p21-22 is low (merely 8.42%), it is considered that p16 is not closely associated with the development of sporadic colorectal cancer. PMID- 11869555 TI - [The establishment of a human pancreatic mixed ductal-endocrine carcinoma cell line]. AB - OBJECTIVE: Establishment of a human pancreatic mixed ductal-endocrine carcinoma cell line. METHODS: A specimen of this type of cancer was obtained from a male patient at the head of pancreas. The tumor tissue was minced into pieces and inoculated subcutaneously in nude mice. With two successive subcutaneous implants, a piece of tumor tissue taken from the 2(nd) implanted neoplasm of a nude mouse was minced into minute tissue masses and inoculated in a media containing 10% fetal bovine serum. RESULTS: Currently, it has been passed to the 55(th) passage. Cells of this cell line (PC-EN) kept simultaneously the morphologic patterns of a pancreatic ductal and endocrine tumor which were verified by histochemistry and immunohistochemistry assays. Some of the PC-EN cells showed presence of neuroendocrine granules by electron microscopy. The growth rate of this cell line was relatively low. Kakryotype analysis showed two cell populations with its major mode of chromosomes numbers as 48 in one, and 56 in the other. Study on K-ras gene demonstrated that PC-EN harbored a mutated codon 12. Several gastrointestinal hormones and CEA were detected in PC-EN cells and in the supra-cultured medium using radioimmunoassay. CONCLUSIONS: The cellular and molecular biologic characteristics of PC-EN cells illustrated that this cell line preserved the morphology of a pancreatic ductal and endocrine tumor and possessed a special growth property, chromosome karyotype, and mutation of Ki-ras gene, but also showed a biologic exocrine and endocrine functions of the pancreas. The establishment of PC-EN cell line will provide a useful model in vitro for further investigating the cellular and molecular biology of exocrine and endocrine tumors of the pancreas. PMID- 11869552 TI - [A clinicopathologic study on eight cases of cystic and solid tumors of pancreas]. AB - OBJECTIVE: To study the clinicopathologic characteristics, differentiation patterns and histogenesis of cystic and solid tumors of the pancreas (CSTP). METHODS: 8 cases of CSTP were studied using histologic (HE and PAS), immunohistochemical (S-P method) and electron microscopic techniques. RESULTS: All the patients were adolescent and young adult females, 14-33 years in age (mean 25.3 years) without recurrence after tumor resection. The mean diameter of tumors was 9.6 cm, all encapsulated. Histological examination showed presence of solid sheets, pseudopapillary, in all of the cases. Hemorrhage, foam cells, and cholesterol crystals were often found. Immunohistochemically, 8 cases were positive for alpha(1)-AT and lysozyme; 6 cases expressed vimentin, 2 cases expressed actin, and CgA-positive cells found in the tumor cell nests in one case. All of the cases showed PR, and 4 cases showed ER positive immunoreactivity in the majority of tumor cells, but negative for CK AE1, CK AE3, EMA, Synaptophysin, ACTH, gastrin, somatostatin, insulin, and glucogan in all the cases. Electron microscopy of 3 cases showed evidences of polymorphism in differentiation of the tumor cells, including the transitional appearance into ducts, acinus, and endocrine cells. Weibel-Palade body found in tumor cells in one out of 8 cases. CONCLUSIONS: (1) CSTP is a distinct clinicopathologic entity in young female patients with a benign clinical course. (2) CSTP develops from primitive pancreatic cells, with the potentiality of developing into ducts, acinus, and endocrine cells. PMID- 11869556 TI - [The expression of integrin alpha5beta1 and transforming growth factor-beta in pulmonary fibrosis of rat]. AB - OBJECTIVE: To study the roles of integrin alpha(5)beta(1) played in rat pulmonary fibrosis. METHODS: The dynamic changes of fibronectin (FN), FN receptor alpha(5)beta(1), and TGF-beta were analysed in bleomycin-induced rat pulmonary fibrosis by using immunohistochemical technique. Northern blot analysis and immunocytochemistry techniques were performed in order to detect the expression of alpha(5)beta(1), FN mRNA and their relevant proteins in cultured rat pulmonary fibroblasts (PFbs) after administration of TGF-beta1. RESULTS: (1) A strong positive reaction with anti-integrin alpha(5)beta(1) was seen in the pneumocytes, endothelial cells and proliferated mesenchymal cells on the 1st to 3rd day after bleomycin. The positive signals for integrin alpha(5)beta(1) were apparent mainly in those proliferated interstitial cells with fibroblast and myofibroblast differentiation 7 days after bleomycin treatment. Changes in FN were similar to that of the integrin alpha(5)beta(1). (2) Expression of alpha(5)beta(1), FN mRNAs and their relevant proteins was increased in rat PFbs after TGF-beta1 administration. CONCLUSIONS: Integrin alpha(5)beta(1) played a key role in inducing the activation, proliferation and differentiation of PFbs, causing an increase of extra-cellular matrix synthesis during pulmonary fibrogenesis. PMID- 11869557 TI - [The effects of transforming growth factor alpha and beta 1 on the proliferation of alveolar type II cells in vitro]. AB - OBJECTIVE: To investigate the effect of transforming growth factor alpha and beta(1) (TGFalpha and beta(1)) on the proliferation of alveolar type II cells in vitro and the molecular mechanism related. METHODS: (3)H-TdR incorporation and cell counting for the assay of cell growth; dot blot, in situ hybridization, and immunohistochemical methods for analysing the expression of cyclin D1 and cycle dependent kinase 4 (CDK4) genes. RESULTS: With increasing of the concentration of TGFalpha and beta(1) (0.01 ng-100 ng/ml), TGFalpha enabled to increase (3)H-TdR incorporation and the number of alveolar type II cells, while the effect of TGFbeta(1) was opposite and both effects were dose-dependent (P < 0.01). TGFalpha treated cells decreased the expression of CDK4 mRNA, and the relevant in comparing with the control (P < 0.01). On the contrary, TGFbeta(1)-treated cells decreased the expression of CDK4 mRNA), CDK4 as well as the cyclin D1 proteins (P < 0.05). CONCLUSION: Both TGFalpha and beta(1) were considered implicated in the regulation of proliferation of cultured alveolar type II cells isolated from the adult rats and CDK4 might be a common target against TGFalpha and beta(1) signals. PMID- 11869558 TI - [V-ATPase inhibitor baflomycine A1 inhibits bone resorption by osteoclast-like cells]. AB - OBJECTIVE: In order to compare the role of osteoclast-like cells (OLCs) originating from giant cell tumor of bone and the osteoclasts (OCs) on bone resorption, as well as to study which kind of proton pump is associated with bone resorption by the OLCs and its role related. METHODS: Culture of OLCs together with the bone slices, the number of pits and the surface areas formed by OLCs was counted. The effects of F0F1-ATPase inhibitor, oligmycine V-ATPase inhibitor, baflomycine of OLCs on bone resorption were compared. RESULTS: Baflomycine are considered effective in inhibiting bone resorption but no definited effect obtained with oligmycine. CONCLUSIONS: OLCs bear the similar bone resorption mechanism as that of OCs. It's considered that OLCs and OCs are derived from the same origin. PMID- 11869559 TI - [The mechanism of bystander effect in herpes simplex virus thymidine kinase/ganciclovir-mediated gene therapy]. AB - OBJECTIVE: To study the mechanism of bystander effect in herpes simplex virus thymidine kinase (HSV-TK)/Ganciclovir (GCV)-mediated gene therapy. METHODS: Recombinant retroviral vectors expressing HSV-TK and beta-Galactosidase (Lac Z) genes were constructed and transferred into pancreatic carcinoma cell line respectively. Cell counting was used to detect the growth inhibition rate of HSV TK-transduced cells in presence of GCV. Taking Lac Z-transduced cells as bystander cells, the bystander effect was detected by MTT method, and its mechanism was studied by the experiments of supernatant shifting, Verapamil inhibition and ultrastructural observation. RESULTS: The growth inhibition rate of the HSV-TK-transduced cells in the presence of GCV was 92.1%, which was obviously 4.9% and 3.2% higher than of the non-and control vector-transduced cells. Mixed cells containing only 10% of HSV-TK-transduced cells showed 39.0% reduction of the proliferation, which meant there was an obvious bystander effect in the system. However this effect disappeared when transferring GCV-containing supernatant of HSV-TK-transduced cells to the parent cells and could be reduced significantly when verapamil was added in the medium, indicating that this bystander effect requires cell-cell contact. Gap junctions were observed existing between PC-2 cells by electron microscopy. CONCLUSION: The bystander effect in HSV-TK/GCV-mediated gene therapy occurs by transfer of GCV metabolite from cell to cell through gap junction. PMID- 11869560 TI - [Expression of endothelin 1 and tumor necrosis factor alpha in injuried renal tubules and their influences on renal interstitial fibroblasts]. AB - OBJECTIVE: To study the expression of endothelin 1 (ET-1) and tumor necrosis factor (TNFalpha) in the epithelial cells of normal and injured renal tubules and their influences on renal interstitial fibroblasts. METHODS: Cultivation of renal tubular epithelial cells and renal interstitial fibroblasts and establishment of a renal tubulo-interstitial fibrosis (TIF) animal model; reverse transcription polymerase chain reaction (RT-PCR), immunohistochemical staining, radioimmunoassay (RIA), double immunohistochemical; and (3)H-TDR incorporation techniques were applied to study the relationship between ET-1, plus TNFalpha and the proliferation of renal interstitial fibroblasts. RESULTS: Expression of ET-1 mRNA and TNFalpha mRNA from renal tubular epithelial cells was obtained (546 bp and 415 bp) respectively as well as the relevant ET-1 and TNFalpha proteins. The concentration of ET-1 and TNFalpha in the culture medium were 1.42 pg/ml and 0.58 ng/ml respectively. The amount of ET-1 and TNFalpha was increased during cell injury and regeneration. In addition, the ratio of (3)H-TDR incorporation and the expression of ET-R mRNA and TNF-R1 mRNA (545 bp and 347 bp) were markedly increased than that of the control group (P < 0.05) when ET-1 or TNFalpha was added in the culture media. CONCLUSION: Injured and regenerated renal tubular epithelial cells are able to synthesize and liberate more ET-1 and TNFalpha than that of the normal renal tubules. ET-1 and TNFalpha are also effective in promoting the proliferation of renal interstitial fibroblasts through ET-R and TNF-R. PMID- 11869561 TI - Amnioinfusion for meconium-stained liquor in labour. AB - BACKGROUND: Amnioinfusion aims to prevent or relieve umbilical cord compression during labour by infusing a solution into the uterine cavity. It is also thought to dilute meconium when present in the amniotic fluid and so reduce the risk of meconium aspiration. However, it may be that the mechanism of effect is that it corrects oligohydramnios (reduced amniotic fluid), for which thick meconium staining is a marker. OBJECTIVES: The objective of this review was to assess the effects of amnioinfusion for meconium-stained liquor on perinatal outcome. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register (October 2001) and the Cochrane Controlled Trials Register (Issue 3, 2001) were searched. SELECTION CRITERIA: Randomised trials comparing amnioinfusion with no amnioinfusion for women in labour with moderate or thick meconium-staining of the amniotic fluid. DATA COLLECTION AND ANALYSIS: Eligibility and trial quality were assessed by one reviewer. MAIN RESULTS: Twelve studies, most involving small numbers of participants, were included. Under standard perinatal surveillance, amnioinfusion was associated with a reduction in the following: heavy meconium staining of the liquor (relative risk 0.03, 95% confidence interval 0.01 to 0.15); variable fetal heart rate deceleration (relative risk 0.65, 95% confidence interval 0.49 to 0.88); and reduced caesarean section overall (relative risk 0.82, 95% confidence interval 0.69 to 1.97). No perinatal deaths were reported. Under limited perinatal surveillance, amnioinfusion was associated with a reduction in the following: meconium aspiration syndrome (relative risk 0.24, 95% confidence interval 0.12 to 0.48); neonatal hypoxic ischaemic encephalopathy (relative risk 0.07, 95% confidence interval 0.01 to 0.56) and neonatal ventilation or intensive care unit admission (relative risk 0.56, 95% confidence interval 0.39 to 0.79); there was a trend towards reduced perinatal mortality (relative risk 0.34, 95% confidence interval 0.11 to 1.06). REVIEWER'S CONCLUSIONS: Amnioinfusion is associated with improvements in perinatal outcome, particularly in settings where facilities for perinatal surveillance are limited. The trials reviewed are too small to address the possibility of rare but serious maternal adverse effects of amnioinfusion. PMID- 11869562 TI - Antidepressants for smoking cessation. AB - BACKGROUND: There are two reasons to believe antidepressants might help in smoking cessation. First, depression may be a symptom of nicotine withdrawal, and smoking cessation sometimes precipitates depression. Second, nicotine may have antidepressant effects that maintain smoking for some smokers. Antidepressants may substitute for this effect. OBJECTIVES: The aim of this review is to assess the effectiveness of antidepressant medications in aiding long term smoking cessation. The drugs include bupropion; doxepin; fluoxetine; imipramine; moclobemide; nortriptyline; paroxetine; selegiline; sertraline, tryptophan and venlafaxine. SEARCH STRATEGY: We searched the Cochrane Tobacco Addiction Group trials register which includes trials indexed in MEDLINE, EMBASE, SciSearch and PsycLIT, and other reviews and meeting abstracts, in September 2001. SELECTION CRITERIA: We considered randomized trials comparing antidepressant drugs to placebo or an alternative therapeutic control for smoking cessation. We excluded trials with less than 6 months follow-up. DATA COLLECTION AND ANALYSIS: We extracted data in duplicate on the type of study population, the nature of the drug therapy, the outcome measures, method of randomisation, and completeness of follow-up. The main outcome measure was abstinence from smoking after at least six months follow-up in patients smoking at baseline. We used the most rigorous definition of abstinence for each trial, and biochemically validated rates if available. Where appropriate, we performed meta-analysis using a fixed effects model. MAIN RESULTS: There was one trial each of moclobemide, sertraline and venlafaxine, two of fluoxetine and nortriptyline, and five trials of bupropion, one of which tested long term use to prevent relapse. Nortriptyline and bupropion both increased cessation. In one trial the combination of bupropion and nicotine patch produced slightly higher quit rates than patch alone. REVIEWER'S CONCLUSIONS: Some antidepressants (bupropion and nortriptyline) can aid smoking cessation. It is not clear whether these effects are specific for individual drugs, or would occur with any antidepressant. PMID- 11869563 TI - Gamma and other cephalocondylic intramedullary nails versus extramedullary implants for extracapsular hip fractures. AB - BACKGROUND: Cephalocondylic intramedullary nails which are inserted proximally to distally (cephalocondylic) have been used for the surgical treatment of extracapsular hip fractures. OBJECTIVES: To compare all cephalocondylic intramedullary nails with extramedullary implants for the surgical treatment of extracapsular hip fractures in adults. This is the third update of our original review which compared the Gamma nail with the sliding hip screw (SHS). SEARCH STRATEGY: We searched the Cochrane Musculoskeletal Injuries Group trials register, MEDLINE, select orthopaedic journals and conference proceedings, and reference lists of relevant articles. We contacted trialists, colleagues and implant manufacturers. Date of the most recent search: August 2001. SELECTION CRITERIA: All randomised and quasi-randomised trials comparing cephalocondylic nails with extramedullary implants for extracapsular hip fractures. DATA COLLECTION AND ANALYSIS: Both reviewers independently assessed trial quality and extracted data. Additional information was sought from all trialists. Wherever appropriate and possible, results were pooled. MAIN RESULTS: The one trial of 230 patients comparing the Kuntscher-Y nail with the SHS, reported no major difference the outcome aside from a significantly increased number of patients with leg shortening, and a tendency for poorer recovery of mobility in the Kuntscher-Y nail group. Seventeen trials comparing the Gamma nail with the SHS were included, with data available for 2472 patients. The Gamma nail was associated with an increased risk of operative and later fracture of the femur and an increased re-operation rate. There were no major differences in the incidence of wound infection, mortality or medical complications between implants. Data were inadequate to determine if there were differences for other outcomes. Five trials involving 603 patients compared the intramedullary hip screw (IMHS) with the SHS. Fracture fixation complications were more common in the IMHS group: all cases of operative and later fracture of the femur occurred in this group. Results for post-operative complications, mortality and functional outcomes were similar in the two groups. Two under-reported trials tested the proximal femoral nail (PFN). The results of one study of 206 patients with a trochanteric fracture showed no advantages for the PFN compared with the SHS. The other study, involving 39 patients, comparing the PFN with the dynamic condylar plate for treating more distal and uncommon trochanteric fractures gave better intra-operative and fracture fixation results for the PFN. One trial of 60 patients reported favourable preliminary results for an experimental mini invasive static intramedullary nail compared with the SHS. REVIEWER'S CONCLUSIONS: Further evidence is required before any conclusions can be drawn on the relative merits of either the Kuntscher-Y nail or the mini-invasive static intramedullary nail in comparison with the SHS. Given the lower complication rate of the SHS in comparison with intramedullary nails, it appears that for trochanteric fractures the SHS is superior. Further studies will be required to determine if different types of intramedullary nail produce the same results, or if intramedullary nails have advantages for selected fracture types, for example, reversed fracture lines and subtrochanteric fractures. From the evidence available, IMHS appears to have the same problems as the Gamma nail, but other theoretical advantages of the IHMS cannot be ruled out. PMID- 11869564 TI - Interventions for reducing the risk of mother-to-child transmission of HIV infection. AB - BACKGROUND: At the end of 1998 over 33 million people were infected with the human immunodeficiency virus (HIV) and over one million children had been infected from their mothers. OBJECTIVES: The objective of this review was to assess what interventions may be effective in decreasing the risk of mother-to child transmission of HIV infection as well as their effect on neonatal and maternal mortality and morbidity. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register were searched. SELECTION CRITERIA: Randomised trials comparing any intervention aimed at decreasing the risk of mother-to-child transmission of HIV infection compared with placebo or no treatment, or any two or more interventions aimed at decreasing the risk of mother-to-child transmission of HIV infection. DATA COLLECTION AND ANALYSIS: Trial quality assessments and data extraction were undertaken by the reviewer. MAIN RESULTS: Zidovudine Four trials comparing zidovudine with placebo involving 1585 participants were included. Compared with placebo, there was a significant reduction in the risk of mother-to-child transmission with any zidovudine (relative risk (RR) 0.54, 95% confidence interval (CI) 0.42-0.69). There is no evidence that 'long course therapy' is superior to 'short course therapy'. Nevirapine One trial compared intrapartum and postnatal nevirapine with intrapartum and postnatal zidovudine in 626 women, the majority of whom breast fed their infants. Compared with zidovudine, there was a significant reduction in the risk of mother-to-child transmission of HIV with nevirapine (RR 0.58, 95% CI 0.40-0.83). No trials are available comparing nevirapine with placebo. Caesarean section One trial comparing elective caesarean section with anticipation of vaginal delivery involving 436 participants was included. Compared with vaginal delivery, there was a significant reduction in the risk of mother-to-child transmission of HIV infection with caesarean section (RR 0.17, 95% CI 0.05-0.55). Immunoglobulin One trial comparing hyperimmune immunoglobulin plus zidovudine with non-specific immunoglobulin plus zidovudine involving 501 participants was included. The addition of hyperimmune immunoglobulin to zidovudine does not appear to have any additional effect on the risk of mother-to-child transmission (RR 0.67, 95% CI 0.29-1.55). REVIEWER'S CONCLUSIONS: Zidovudine, nevirapine and delivery by elective caesarean section appear to be very effective in decreasing the risk of mother-to-child transmission of HIV infection. PMID- 11869565 TI - Interventions for leg cramps in pregnancy. AB - BACKGROUND: Many women experience leg cramps in pregnancy. They become more common as pregnancy progresses and are especially troublesome at night. OBJECTIVES: The objective of this review was to assess methods of preventing and treating leg cramps in pregnancy. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register (October 2001). SELECTION CRITERIA: Randomised trials of treatments for leg cramps in pregnancy. DATA COLLECTION AND ANALYSIS: Trial quality was assessed and data were extracted independently by two reviewers. MAIN RESULTS: Five trials involving 352 women were included. The trials were of moderate quality. The only placebo-controlled trial of calcium treatment showed no evidence of benefit. Trials comparing sodium chloride with placebo (odds ratio 0.54, 95% confidence interval 0.23 to 1.29) and calcium with sodium chloride (odds ratio 1.23, 95% confidence intervals 0.47 to 3.27 ) showed no evidence of benefit. Placebo controlled trials of multivitamin with mineral supplements (odds ratio 0.23, 95% confidence intervals 0.05 to 1.01) and magnesium (odds ratio 0.18, 95% confidence intervals 0.05 to 0.60) provided some suggestion of benefit. REVIEWER'S CONCLUSIONS: The evidence that calcium reduces cramp is weak and seems to depend on placebo effect. The evidence for sodium chloride is stronger but the results of the sodium chloride trial may no longer be relevant because of dietary changes which include an increased sodium intake in the general population. It is not possible to recommend multivitamins with mineral supplementation, as it is not clear which ingredient, if any, is helping. If a woman finds cramp troublesome in pregnancy, the best evidence is for magnesium lactate or citrate taken as 5mmol in the morning and 10mmol in the evening. PMID- 11869566 TI - Maternal hydration for increasing amniotic fluid volume in oligohydramnios and normal amniotic fluid volume. AB - BACKGROUND: Oligohydramnios (reduced amniotic fluid) may be responsible for malpresentation problems, umbilical cord compression, concentration of meconium in the liquor, and difficult or failed external cephalic version. Simple maternal hydration has been suggested as a way of increasing amniotic fluid volume in order to reduce some of these problems. OBJECTIVES: The objective of this review was to assess the effects of maternal hydration on amniotic fluid volume and measures of pregnancy outcome. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register were searched. Date of last search: September 2001. SELECTION CRITERIA: Randomised trials comparing maternal hydration with no hydration in pregnant women with reduced or normal amniotic fluid volume. DATA COLLECTION AND ANALYSIS: Eligibility and trial quality were assessed by both reviewers. MAIN RESULTS: Two studies of 77 women were included. The women were asked to drink two litres of water before having a repeat ultrasound examination. Maternal hydration in women with and without oligohydramnios was associated with an increase in amniotic volume (weighted mean difference for women with oligohydramnios 2.01, 95% confidence interval 1.43 to 2.56; and weighted mean difference for women with normal amniotic fluid volume 4.5, 95% confidence interval 2.92 to 6.08). Intravenous hypotonic hydration in women with oligohydramnios was associated with an increase in amniotic fluid volume (weighted mean difference 2.3, 95% confidence interval 1.36 to 3.24). Isotonic intravenous hydration had no measurable effect. No clinically important outcomes were assessed in any of the trials. REVIEWER'S CONCLUSIONS: Simple maternal hydration appears to increase amniotic fluid volume and may be beneficial in the management of oligohydramnios and prevention of oligohydramnios during labour or prior to external cephalic version. Controlled trials are needed to assess the clinical benefits and possible risks of maternal hydration for specific clinical purposes. PMID- 11869567 TI - Interventions for nausea and vomiting in early pregnancy. AB - BACKGROUND: Nausea and vomiting are the most common symptoms experienced in early pregnancy, with nausea affecting between 70 and 85% of women. About half of pregnant women experience vomiting. OBJECTIVES: The objective of this review was to assess the effects of different methods of treating nausea and vomiting in early pregnancy. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register. Date of last search: October 2001. SELECTION CRITERIA: Randomised trials of any treatment for nausea and/or vomiting in early pregnancy. DATA COLLECTION AND ANALYSIS: Trial quality was assessed and data were extracted independently by two reviewers. MAIN RESULTS: Twenty-three trials were included. These trials were of variable quality. Nausea treatments were different anti-histamine medications, vitamin B6 (pyridoxine), the combination tablet Debendox (Bendectin) and P6 acupressure. For hyperemesis gravidarum five trials were identified testing treatments with oral ginger root extract, oral corticosteroids or injected adrenocorticotropic hormone (ACTH) and intravenous diazepam. Based on 13 trials, there was an overall reduction in nausea from anti-emetic medication (odds ratio 0.17, 95% confidence interval 0.13 to 0.21). REVIEWER'S CONCLUSIONS: Anti-emetic medication appears to reduce the frequency of nausea in early pregnancy. There is some evidence of adverse effects, but there is very little information on effects on fetal outcomes from randomised controlled trials. Of newer treatments, pyridoxine (vitamin B6) appears to be more effective in reducing the severity of nausea. The results from trials of P6 acupressure are equivocal. No trials of treatments for hyperemesis gravidarum show any evidence of benefit. Evidence from observational studies suggests no evidence of teratogenicity from any of these treatments. PMID- 11869568 TI - Steroid sex hormones for lower limb atherosclerosis. AB - BACKGROUND: There is accumulating evidence that steroid sex hormones have a beneficial effect on a number of risk factors for peripheral arterial disease. OBJECTIVES: The objective of this review was to determine whether exogenous steroid sex hormones are an effective treatment for patients with lower limb atherosclerosis. SEARCH STRATEGY: The Cochrane Peripheral Vascular Diseases Group trials register was searched, together with reference lists from relevant articles and reviews obtained through searches of Embase and Medline (up to May 2001). SELECTION CRITERIA: Randomised or quasi-randomised controlled trials of steroid sex hormones in patients with lower limb atherosclerosis were selected. DATA COLLECTION AND ANALYSIS: Both reviewers extracted data and assessed trial quality independently. Whenever possible investigators were contacted to obtain information needed for the review that could not be found in published reports. MAIN RESULTS: Four trials appeared to meet the inclusion criteria, but one was excluded because of poor methodology. The three remaining trials compared testosterone treatment with placebo in a total of 109 subjects with intermittent claudication or critical leg ischaemia. The most recent trial to meet the inclusion criteria dated from 1971. No trials were available which investigated the potentially beneficial effects of oestrogenic hormones in women with lower limb atherosclerosis. Testosterone therapy produced no significant improvement in tests of walking distance or in a variety of other objective tests for peripheral arterial disease, including venous filling time, muscle blood flow and plethysmography. The relative risk for subjective improvement in symptoms using the combined trial results was also non-significant (relative risk 1.10, 95% confidence interval 0.81 to 1.48). REVIEWER'S CONCLUSIONS: There is no evidence to date that short-term testosterone treatment is beneficial in subjects with lower limb atherosclerosis. However, this might reflect limited data rather than the lack of a real effect. PMID- 11869569 TI - Somatostatin analogues for acute bleeding oesophageal varices. AB - BACKGROUND: Somatostatin and its derivatives are often used for emergency treatment of bleeding oesophageal varices in patients with cirrhosis of the liver. The placebo controlled trials have shown varying results, however, and their power has been quite low. An updated systematic review of a previously published meta-analysis was therefore performed. OBJECTIVES: To study whether somatostatin or analogues improve survival or reduce the number of blood transfusions in patients with suspected or verified acute or recently bleeding oesophageal varices. SEARCH STRATEGY: MEDLINE and The Cochrane Library were searched in August 2001. Reference lists of articles and authors. SELECTION CRITERIA: All randomised trials comparing somatostatin or analogues with placebo or no treatment in patients suspected of acute bleeding from oesophageal varices. DATA COLLECTION AND ANALYSIS: The effect variables extracted were: mortality, number of blood transfusions, number with balloon tamponade, number with initial haemostasis and number with rebleeding. Intention to treat analyses were conducted; a random effects analysis was preferred if there was significant heterogeneity between the trials (P < 0.10). MAIN RESULTS: The meta-analysis comprised 12 trials and 1452 patients. The active drugs did not reduce mortality significantly (relative risk 0.93, 95% confidence interval (CI) 0.75 to 1.14). The number of transfusions was less with drugs, the difference between experimental and control therapy was 1.0 units of blood products (95% CI 0.6 to 1.5). Number of patients failing initial haemostasis was also reduced (relative risk 0.68, 95% CI 0.50 to 0.92). There were no significant differences in use of balloon tamponade (relative risk 0.68, 95% CI 0.37 to 1.24), or in number of patients with rebleeding (relative risk 0.61, 95% CI 0.35 to 1.09). It should be noted, however, that the trials were heterogeneous with respect to the secondary outcomes. REVIEWER'S CONCLUSIONS: The effect corresponded to one unit of blood saved per patient. It can be discussed whether this effect is worthwhile. The findings do not suggest a need for further placebo-controlled studies of somatostatin analogues in acute bleeding from oesophageal varices. On the other hand, the confidence interval for the effect on mortality was wide. Hence, a large placebo controlled trial is needed if one wishes to rule out the possibility that a worthwhile effect on mortality may have been overlooked. PMID- 11869570 TI - Organised inpatient (stroke unit) care for stroke. AB - BACKGROUND: Organised stroke unit care is provided by multidisciplinary teams that exclusively manage stroke patients in a dedicated ward (stroke ward), with a mobile team (stroke team) or within a generic disability service (a mixed rehabilitation ward). OBJECTIVES: The objective of this review was to assess the effect of stroke unit care compared with alternative forms of care for patients following a stroke. SEARCH STRATEGY: We searched the Cochrane Stroke Group trials register, reference lists of articles and contacted researchers in the field. SELECTION CRITERIA: Randomised and quasi-randomised trials comparing organised inpatient stroke unit care with an alternative service. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed eligibility and trial quality. The principal reviewer conducted a structured interview with the coordinators of unpublished trials. MAIN RESULTS: Twenty three trials were included. Compared with alternative services, stroke unit care showed reductions in the odds of death recorded at final (median one year) follow-up (odds ratio 0.86; 95% confidence interval 0.71 to 0.94; P=0.005), the odds of death or institutionalised care (0.80; 0.71 to 0.90; P=0.0002) and death or dependency (0.78; 0.68 to 0.89; P=0.0003). Subgroup analyses indicated that the observed benefits remained when the analysis was restricted to truly randomised trials with blinded outcome assessment. Outcomes were independent of patient age, sex and stroke severity but appeared to be better in stroke units based in a discrete ward. There was no indication that organised stroke unit care resulted in increased hospital stay. REVIEWER'S CONCLUSIONS: Stroke patients who receive organised inpatient care in a stroke unit are more likely to be alive, independent, and living at home one year after the stroke. The benefits were most apparent in units based in a discrete ward. No systematic increase was observed in the length of inpatient stay. PMID- 11869571 TI - Caregiver support for women during childbirth. AB - BACKGROUND: Social support may include advice or information, tangible assistance and emotional support. OBJECTIVES: The objective of this review was to assess the effects of continuous support during labour (provided by health care workers or lay people) on mothers and babies. SEARCH STRATEGY: I searched the Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register. Date of last search: October 2001. SELECTION CRITERIA: Randomised trials comparing continuous support during labour with usual care. DATA COLLECTION AND ANALYSIS: Trial quality was assessed. Study authors were contacted for additional information. MAIN RESULTS: Fourteen trials, involving more than 5000 women, are included in the review. The continuous presence of a support person reduced the likelihood of medication for pain relief, operative vaginal delivery, caesarean delivery, and a 5-minute Apgar score less than 7. Continuous support was also associated with a slight reduction in the length of labour. Six trials evaluated the effects of support on mothers' views of their childbirth experiences; while the trials used different measures (overall satisfaction, failure to cope well during labour, finding labour to be worse than expected, and level of personal control during childbirth), in each trial the results favoured the group who had received continuous support. REVIEWER'S CONCLUSIONS: Continuous support during labour from caregivers (nurses, midwives or lay people) appears to have a number of benefits for mothers and their babies and there do not appear to be any harmful effects. PMID- 11869572 TI - Antioxidant vitamin and mineral supplements for age-related macular degeneration. AB - BACKGROUND: It has been proposed that antioxidants may prevent cellular damage in the retina by reacting with free radicals produced in the process of light absorption. OBJECTIVES: The objective of this review is to assess the effects of antioxidant vitamin and/or mineral supplementation on the progression of age related macular degeneration. SEARCH STRATEGY: The Cochrane Controlled Trials Register - CENTRAL/CCTR, which contains the Cochrane Eyes and Vision Group specialised register (Cochrane Library Issue 3 2001), MEDLINE (1966 to August 2001), EMBASE (1980 to September 2001), the Science Citation Index, and the reference lists of relevant articles were searched. Investigators of included studies were contacted for further information. SELECTION CRITERIA: Randomised trials comparing an antioxidant vitamin and/or mineral supplement (alone or in combination) to control in people with age-related macular degeneration are included in this review. DATA COLLECTION AND ANALYSIS: The reviewer extracted data and assessed trial quality. Due to the variable methods of collecting and presenting outcome data, no statistical summary measure was calculated. MAIN RESULTS: Seven trials, which randomised 4119 people with signs of age-related macular degeneration, are included in this review. One unpublished trial of zinc supplementation (170 participants) is awaiting assessment. The majority of people (88%) were randomised in one trial that was conducted in a relatively well nourished American population. This trial found a modest beneficial effect of antioxidant and zinc supplementation on progression to advanced age-related macular degeneration (odds ratio 0.72, 99% confidence interval 0.52 to 0.98). People supplemented with antioxidants and zinc were less likely to lose 15 or more letters of visual acuity (equivalent to a doubling of the visual angle) (odds ratio 0.79, 99% confidence interval 0.60 to 1.04). The other six trials in this review were small and the results were inconsistent. REVIEWER'S CONCLUSIONS: The evidence as to the effectiveness of antioxidant vitamin and mineral supplementation in halting the progression of age-related macular degeneration is dominated by one large trial in a relatively well-nourished American population that showed modest benefit in people with moderate to severe signs of the disease. There is no evidence at present that people with early signs of the disease should take supplementation, however, current studies are underpowered to answer that question. The generalisability of these findings to other populations with different nutritional status is not known. Further large well-conducted randomised controlled trials in other populations are required. PMID- 11869573 TI - Interferon for acute hepatitis C. AB - BACKGROUND: Acute hepatitis C virus (HCV) infection progresses to chronicity in the majority of patients. In order to prevent the progression to chronic disease, several studies have assessed interferon in patients with acute hepatitis C. OBJECTIVES: The aim of this review was to assess the efficacy of interferon in acute HCV infection. SEARCH STRATEGY: We searched MEDLINE, the Cochrane Controlled Trials Register, and the abstracts of the American Association for the Study of Liver Diseases (June 2001). We also contacted pharmaceutical companies to obtain unpublished trials. SELECTION CRITERIA: Randomised clinical trials comparing interferon with placebo or no treatment, and published as an article, abstract, or letter were selected. No language limitations were used. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed trial quality and extracted data. The following endpoints were analysed: normalization of alanine aminotransferase (ALT) activity at the end of treatment (biochemical ETR); sustained ALT normalization at the end follow-up (biochemical SR); disappearance of serum HCV RNA by polymerase chain reaction assay at the end of treatment (virologic ETR) and at the end of follow-up (virologic SR). Histologic data and adverse events were also recorded. Assessment of drug efficacy used the methods of Peto and Der Simonian and Laird. MAIN RESULTS: Six randomised trials involving 206 patients with acute hepatitis C met the inclusion criteria. Four trials assessing interferon alfa-2b in 141 patients, all with transfusion-acquired acute hepatitis C, were included. They demonstrated no significant heterogeneity in the outcomes assessed. When compared with no treatment, interferon alfa-2b was associated with an increase in the rates of virologic ETR and SR by 45% (95% CI 31-59%, P < 0.00001) and 29% (95% CI 14-44%, P = 0.0002), respectively. The virologic ETR was 42% (95% CI: 30-56%) in the interferon alfa-2b group versus 4% (95% CI 0-13%, P < 0.00001) in the control group. At the end of follow-up, a virologic SR was seen in 32% (95% CI 21-46%) of interferon-treated patients versus only 4% (95% CI 0-13%, P = 0.00007) of controls. The tolerability of therapy, or the impact of interferon alfa-2b on hepatic histology, was not reported. Two trials assessed interferon beta in a total 65 patients. The efficacy of interferon beta could not be assessed, however, due to heterogeneity of these trials. REVIEWER'S CONCLUSIONS: Interferon alfa is effective in improving biochemical outcomes and achieving sustained virologic clearance in patients with transfusion-acquired acute hepatitis C. The effect on long-term clinical outcomes could not be assessed due to limitations in the current data. PMID- 11869574 TI - Mass media interventions: effects on health services utilisation. AB - BACKGROUND: The mass media frequently cover health related topics, are the leading source of information about important health issues, and are targeted by those who aim to influence the behaviour of health professionals and patients. OBJECTIVES: To assess the effects of mass media on the utilisation of health services. SEARCH STRATEGY: We searched the Cochrane Effective Practice and Organisation of Care Group specialised register (1996 to 1999), MEDLINE, EMBASE, Eric, PsycLit (to 1999), and reference lists of articles. We hand searched the journals Communication Research (February 1987 to August 1996), European Journal of Communication (1986 to 1994), Journal of Communication (winter 1986 to summer 1996), Communication Theory (February 1991 to August 1996), Critical Studies in Mass Communication (March 1984 to March 1995) and Journalism Quarterly (1986 to summer 1996). SELECTION CRITERIA: Randomised trials, controlled clinical trials, controlled before-and-after studies and interrupted time series analyses of mass media interventions. The participants were health care professionals, patients and the general public. DATA COLLECTION AND ANALYSIS: Two reviewers independently extracted data and assessed study quality. MAIN RESULTS: Twenty studies were included. All used interrupted time series designs. Fifteen evaluated the impact of formal mass media campaigns, and five of media coverage of health-related issues. The overall methodological quality was variable. Six studies did not perform any statistical analysis, and nine used inappropriate statistical tests (ie not taking into account the effect of time trend). All of the studies apart from one concluded that mass media was effective. These positive findings were confirmed by our re-analysis in seven studies. The direction of effect was consistent across studies towards the expected change. REVIEWER'S CONCLUSIONS: Despite the limited information about key aspects of mass media interventions and the poor quality of the available primary research there is evidence that these channels of communication may have an important role in influencing the use of health care interventions. Although the findings of this review may be affected by publication bias, those engaged in promoting better uptake of research information in clinical practice should consider mass media as one of the tools that may encourage the use of effective services and discourage those of unproven effectiveness. PMID- 11869575 TI - Nonsteroidal anti-inflammatory drugs for heavy menstrual bleeding. AB - BACKGROUND: Heavy menstrual bleeding (HMB) is an important cause of ill health in premenopausal women. Although surgery is often used as a treatment, a range of medical therapies are also available. Nonsteroidal anti-inflammatory drugs reduce prostaglandin levels which are elevated in women with excessive menstrual bleeding and also may have a beneficial effect on dysmenorrhoea. OBJECTIVES: The primary objective of this review was to investigate the effectiveness of non steroidal anti-inflammatory drugs (NSAIDs) in achieving a reduction in menstrual blood loss in women of reproductive years HMB. SEARCH STRATEGY: Electronic searches for relevant randomised controlled trials of the Cochrane Menstrual Disorders and Subfertility Group Specialised Register of controlled trials, MEDLINE, EMBASE, Current Contents, the Cochrane Library and CINAHL were performed. Attempts were also made to identify trials from citation lists of review articles and drug companies were approached for unpublished data. In most cases, the first author of each included trial was contacted for additional information. An updated search was performed in September and October 2001 but no new eligible trials were identified. SELECTION CRITERIA: The inclusion criteria were randomised comparisons of individual NSAIDs with either each other, placebo or other medical treatments in women with regular heavy periods measured either objectively or subjectively and with no pathological or iatrogenic (treatment induced) causes for their heavy menstrual blood loss. DATA COLLECTION AND ANALYSIS: Sixteen RCTs were identified that fulfilled the inclusion criteria for this review and data were extracted independently. Odds ratios for dichotomous outcomes and weighted mean differences for continuous outcomes were estimated from the data of nine trials. The results of the remaining seven crossover trials with data unsuitable for pooling were described in the Other Data section. MAIN RESULTS: As a group, NSAIDs were more effective than placebo at reducing heavy menstrual bleeding but less effective than either tranexamic acid or danazol. Treatment with danazol caused a shorter duration of menstruation and more adverse events than NSAIDs but this did not appear to affect the acceptability of treatment. There were no statistically significant differences between NSAIDs and the other treatments (oral luteal progestogen, ethamsylate, progesterone releasing intra-uterine system (IUS), oral contraceptive pill (OCC)) but most studies were underpowered. There was no evidence of a difference between the individual NSAIDs (naproxen and mefenamic acid) in reducing HMB. REVIEWER'S CONCLUSIONS: NSAIDs reduce HMB when compared with placebo but are less effective than either tranexamic acid or danazol. However, adverse events are more severe with danazol therapy. In the limited number of small studies suitable for evaluation, no significant difference in efficacy was demonstrated between NSAIDs and other medical treatments such as oral luteal progestogen, ethamsylate, OCC or IUS. PMID- 11869576 TI - Services for reducing duration of hospital care for acute stroke patients. AB - BACKGROUND: Stroke patients conventionally receive a substantial part of their rehabilitation in hospital. Services have now been developed which offer patients in hospital an early discharge with rehabilitation at home (early supported discharge, ESD). OBJECTIVES: To establish the costs and effects of ESD services compared with conventional services. SEARCH STRATEGY: The Stroke Group Specialist Register of Controlled Trials was searched and supplemented with information from individual trialists. Searching was completed in December 2000. SELECTION CRITERIA: Randomised controlled trials recruiting stroke patients in hospital to receive either conventional care or any service intervention which has provided rehabilitation and support in a community setting with an aim of reducing the duration of hospital care. DATA COLLECTION AND ANALYSIS: Two reviewers scrutinised trials and categorised them on their eligibility. Standardised information was then obtained from the primary trialists. Results were analysed for all trials and for subgroups depending on whether the intervention was provided by a coordinated multidisciplinary team (coordinated ESD team) or not. MAIN RESULTS: Outcome data are currently available for four trials. Patients tended to be a selected elderly group with disability. Overall, the odds ratios (95% confidence interval) for death, death or institutionalisation, death or dependency at the end of scheduled follow up were 0.87 (0.39-1.93), 0.69 (0.36 1.31) and 0.88 (0.49-1.57) respectively. Apparent benefits were more evident in the three trials evaluating a coordinated ESD team. The ESD group showed significant reductions (P<0.001) in the length of hospital stay equivalent to approximately nine days. REVIEWER'S CONCLUSIONS: ESD services provided for a selected group of stroke patients can reduce the length of hospital stay. However, the relative risks and benefits and overall costs of such services remain unclear. PMID- 11869577 TI - Multi-agent chemotherapy for early breast cancer. AB - BACKGROUND: There have been many randomised trials of adjuvant prolonged polychemotherapy among women with early breast cancer, and an updated overview of their results is presented. OBJECTIVES: In this report, the Early Breast Cancer Trialists' Collaborative Group present their updated systematic overview (meta analysis) of treatment with polychemotherapy. SEARCH STRATEGY: Trial identification procedures for the EBCTCG overviews have been described elsewhere. See under "EBCTCG" in the Breast Cancer Collaborative Review Group module. SELECTION CRITERIA: All randomised trials that began before 1990 and involved treatment groups that differed only with respect to the chemotherapy regimens that were being compared. DATA COLLECTION AND ANALYSIS: In 1995, information was sought on each woman in any randomised trial that began before 1990 and involved treatment groups that differed only with respect to the chemotherapy regimens that were being compared. Analyses involved about 18,000 women in 47 trials of prolonged polychemotherapy versus no chemotherapy, about 6000 in 11 trials of longer versus shorter polychemotherapy, and about 6000 in 11 trials of anthracycline-containing regimens versus CMF (cyclophosphamide, methotrexate, and fluorouracil). MAIN RESULTS: For recurrence, polychemotherapy produced substantial and highly significant proportional reductions both among women aged under 50 at randomisation (35% [SD 4] reduction; 2p <0.00001) and among those aged 50-69 (20% [SD 3] reduction; 2p <0.00001); few women aged 70 or over had been studied. For mortality, the reductions were also significant both among women aged under 50 (27% [SD 5] reduction; 2p <0.00001) and among those aged 50 69 (11% [SD 3] reduction; 2p=0.0001). The recurrence reductions emerged chiefly during the first 5 years of follow-up, whereas the difference in survival grew throughout the first 10 years. After standardisation for age and time since randomisation, the proportional reductions in risk were similar for women with node-negative and node-positive disease. Applying the proportional mortality reduction observed in all women aged under 50 at randomisation would typically change a 10-year survival of 71% for those with node-negative disease to 78% (an absolute benefit of 7%), and of 42% for those with node-positive disease to 53% (an absolute benefit of 11%). The smaller proportional mortality reduction observed in all women aged 50-69 at randomisation would translate into smaller absolute benefits, changing a 10-year survival of 67% for those with node negative disease to 69% (an absolute gain of 2%) and of 46% for those with node positive disease to 49% (an absolute gain of 3%). The age-specific benefits of polychemotherapy appeared to be largely irrespective of menopausal status at presentation, oestrogen receptor status of the primary tumour, and of whether adjuvant tamoxifen had been given. In terms of other outcomes, there was a reduction of about one-fifth (2p=0.05) in contralateral breast cancer, which has already been included in the analyses of recurrence, and no apparent adverse effect on deaths from causes other than breast cancer (death rate ratio 0.89 [SD 0.09]). The directly randomised comparisons of longer versus shorter durations of polychemotherapy did not indicate any survival advantage with the use of more than about 3-6 months of polychemotherapy. By contrast, directly randomised comparisons did suggest that, compared with CMF alone, the anthracycline containing regimens studied produced somewhat greater effects on recurrence (2p=0.006) and mortality (69% vs 72% 5-year survival; log-rank 2p=0.02). But this comparison is one of many that could have been selected for emphasis, the 99% CI reaches zero, and the results of several of the relevant trials are not yet available. REVIEWER'S CONCLUSIONS: Some months of adjuvant polychemotherapy (eg, with CMF or an anthracycline-containing regimen) typically produces an absolute improvement of about 7-11% in 10-year survival for women aged under 50 at presentation with early breast cancer, and of about 2-3% for those aged 50-69 (unless their prognosis is likely to be extremely good even without such treatment). Treatment decisions involve consideration not only of improvements in cancer recurrence and survival but also of adverse side-effects of treatment, and this report makes no recommendations as to who should or should not be treated. PMID- 11869578 TI - Nebulized racemic epinephrine for extubation of newborn infants. AB - BACKGROUND: Following a period of mechanical ventilation, post-extubation upper airway obstruction can occur in newborn infants, especially after prolonged, traumatic or multiple intubations. The subsequent increase in upper airway resistance may lead to respiratory insufficiency and failure of extubation. The vasoconstrictive properties of epinephrine, and its proven efficacy in the treatment of croup in infants, has led to the routine use of inhaled nebulized epinephrine immediately post-extubation in some neonatal units. It is also recommended for neonates with post-extubation tracheal obstruction and stridor in neonatal and respiratory textbooks and reviews. OBJECTIVES: The primary objective was to assess whether nebulized epinephrine administered immediately after extubation in neonates weaned from IPPV decreases the need for subsequent additional respiratory support. SEARCH STRATEGY: Searches were of: MEDLINE from 1966 to September 2000; CINAHL from 1982 to September 2000; Current Contents from 1994 to September 2000; and the Cochrane Controlled Trials Register (Cochrane Library Issue 3, 2000). These searches were updated to September 2001 for this review update. Previous searches up to March 1999 included the Oxford Database of Perinatal Trials, expert informants and journal hand searching mainly in the English language, previous reviews including cross references, abstracts, and conference and symposia proceedings. SELECTION CRITERIA: All randomised and quasi randomised control trials in which nebulized epinephrine was compared with placebo immediately post-extubation in newborn infants who have been weaned from IPPV and extubated, with regard to clinically important outcomes (i.e. need for additional respiratory support, increase in oxygen requirement, respiratory distress, stridor or the occurrence of side effects). DATA COLLECTION AND ANALYSIS: No studies met our criteria for inclusion in this review. MAIN RESULTS: No studies were identified which looked at the effect of inhaled nebulized epinephrine on clinically important outcomes in infants being extubated. REVIEWER'S CONCLUSIONS: IMPLICATIONS FOR PRACTICE: There is no evidence either supporting or refuting the use of inhaled nebulized racemic epinephrine in newborn infants. IMPLICATIONS FOR RESEARCH: randomised controlled trials are needed comparing inhaled nebulized racemic epinephrine with placebo in neonates post-extubation. This should be looked at both as a routine treatment post extubation and as specific treatment for post-extubation upper airway obstruction. Study populations should include the group of infants at highest risk for upper airway obstruction from mucosal swelling because of their small glottic and sub-glottic diameters (ie those infants with birthweights less than 1000 grams). PMID- 11869579 TI - Cognitive behaviour therapy for schizophrenia. AB - BACKGROUND: Medication is the mainstay of treatment for schizophrenia. Many people with schizophrenia, however, continue to experience symptoms in spite of medication and may experience side effects that are unwanted and unpleasant. In addition to medication additional forms of treatment include talking therapies such as cognitive behavioural therapy. This approach helps to link the person's feelings and patterns of thinking which underpin distress. OBJECTIVES: To review the effectiveness of cognitive behavioural therapy for people with schizophrenia, when compared to standard care, specific medication, other therapies and non intervention. SEARCH STRATEGY: Electronic searches of Biological Abstracts (1980 1998), CINAHL (1982-1998), The Cochrane Library (Issue 2, 1998), The Cochrane Schizophrenia Groups' Register of Trials, which encompasses up to date searches of all listed databases (January 2001), EMBASE (1980-1998), MEDLINE (1966-1998), PsychLIT (1887-1997), SIGLE (1990-1998), Sociofile (1980-1998) were undertaken. All references of the articles selected were searched for further relevant trials. SELECTION CRITERIA: This review includes relevant randomised trials of cognitive behaviour therapy for people with a diagnosis of schizophrenia-like illnesses. Outcomes such as death, mental state, relapse, psychological well being and acceptability of treatment were sought. DATA COLLECTION AND ANALYSIS: Studies were reliably selected and assessed for methodological quality. Data were extracted by two reviewers working independently. Dichotomous data were analysed on an intention-to-treat basis and continuous data with 65% completion rate are presented. Where possible, for dichotomous outcomes, a relative risk (RR) with the 95% confidence interval (CI) was estimated along with the number needed to treat statistic (NNT). MAIN RESULTS: Twenty-two relevant papers describing thirteen trials were identified. Cognitive behavioural therapy in addition to standard care did not significantly reduce the rate of relapse and readmission to hospital when compared with standard care alone (medium term 1 RCT, N=61, RR 0.1 CI 0.01 to 1.7; long term 2 RCTs, N=123, RR 1.1 CI 0.8 to 1.5). A significant difference was observed, however, favouring cognitive behavioural therapy over standard care alone, in terms of being able to be discharged from hospital (1 RCT, N=62, RR 0.5 CI 0.3 to 0.9, NNT 3 CI 2 to 12). For 'no important improvement in mental state' data showed a significant difference favouring the cognitive behavioural therapy group over standard care alone when measured at 13 to 26 weeks (2 RCTs, N=123, RR 0.7 CI 0.6 to 0.9, NNT 4 CI 2 to 8). After one year the difference was no longer significant (3 RCTs, N=211, RR 0.95 CI 0.6 to 1.5). On continuous measures (BPRS, CPRS, Psychiatric Assessment Scale) data are not convincing of an effect. A cognitive behavioural therapy approach focusing on compliance may have some effects on insight and attitudes to medication, but the clinical meaning of these data is unclear. When compared with supportive psychotherapy, cognitive behavioural therapy had no effects on relapse rate and clinically meaningful improvements in mental state. Cognitive behavioural therapy combined with other psycho-social/educational interventions may decrease the numbers of people able to tolerate the intervention, at least under study conditions. REVIEWER'S CONCLUSIONS: Cognitive behavioural therapy is a promising but under evaluated intervention. Currently, trial-based data supporting the wide use of cognitive behavioural therapy for people with schizophrenia or other psychotic illnesses are far from conclusive. More trials are justified, especially in comparison with a lower grade supportive approach. These trials should be designed to be both clinically meaningful and widely applicable. PMID- 11869580 TI - Ursodeoxycholic acid for primary biliary cirrhosis. AB - BACKGROUND: Primary biliary cirrhosis is a rare autoimmune liver disease and an effective treatment has been difficult to establish. Some randomised clinical trials have found an effect of ursodeoxycholic acid for primary biliary cirrhosis. OBJECTIVES: Evaluate the beneficial effects and adverse effects of peroral ursodeoxycholic acid for primary biliary cirrhosis versus placebo or no intervention. SEARCH STRATEGY: The Controlled Trials Register of The Cochrane Hepato-Biliary Group, The Cochrane Library, MEDLINE, EMBASE and the full text of the identified studies were searched until April 2001. The electronic searches were done by entering the search terms 'ursodeoxycholic acid', 'UDCA', 'primary biliary cirrhosis', and 'PBC'. SELECTION CRITERIA: Randomised clinical trials evaluating ursodeoxycholic acid administered perorally at any dose versus placebo or no intervention in patients with primary biliary cirrhosis diagnosed by any method. Only trials using an adequate method for randomisation were included, regardless of blinding and language. DATA COLLECTION AND ANALYSIS: The methodologic quality of the randomised clinical trials was evaluated by components and the Jadad-score. The following outcomes were extracted: mortality, liver transplantation, pruritus, other clinical symptoms (jaundice, portal pressure, (bleeding) oesophageal varices, ascites, hepatic encephalopathy, hepato renal syndrome, autoimmune conditions), liver biochemistry, liver function, liver biopsy findings, quality of life, and adverse events. All analyses were performed according to the intention-to-treat method. MAIN RESULTS: A total of 16 randomised clinical trials evaluating ursodeoxycholic acid against placebo (n = 15) or no intervention (n = 1) in 1422 patients were identified. The median Jadad score was 3 (range 1-5). A number of trials described as double blind had problems with the blinding. Neither mortality (odds ratio = 0.94; 95% confidence interval (CI) 0.60 to 1.48), liver transplantation (odds ratio = 0.83; 95% CI 0.52 to 1.32), mortality or liver transplantation (odds ratio = 0.90; 95% CI 0.65 to 1.26), pruritus, fatigue, autoimmune conditions, quality of life, liver histology, or portal pressure were significantly affected by ursodeoxycholic acid (given in doses of 8-15 mg/kg/day for three months to five years). However, ursodeoxycholic acid significantly (P < 0.05) reduced ascites, jaundice, and biochemical variables such as serum bilirubin and liver enzymes. Ursodeoxycholic acid was not significantly associated with adverse events. Including data after patients had been switched onto open label ursodeoxycholic acid confirmed the findings regarding the lack of a significant effect of ursodeoxycholic acid on mortality and mortality or liver transplantation. A significant (P = 0.04) effect was, however, observed on the incidence of liver transplantation (odds ratio = 0.68; 95% CI 0.48 to 0.98). REVIEWER'S CONCLUSIONS: Ursodeoxycholic acid has a marginal therapeutic effect for primary biliary cirrhosis. On the positive side, ursodeoxycholic acid has few side effects. The general usage of ursodeoxycholic acid for primary biliary cirrhosis needs reevaluation. PMID- 11869581 TI - Multidisciplinary bio-psycho-social rehabilitation for chronic low back pain. AB - BACKGROUND: Chronic low back pain is, in many countries, the main cause of long term disability in middle age. Patients with chronic low back pain are often referred for multidisciplinary treatment. Previous published systematic reviews on this topic included no randomised controlled trials and pooled together controlled and non-controlled studies. OBJECTIVES: To assess the effect of multidisciplinary bio-psycho-social rehabilitation on pain, function, employment, quality of life and global assessment outcomes in subjects with chronic disabling low back pain. SEARCH STRATEGY: We searched MEDLINE, EMBASE, PsychLIT, CINAHL, Health STAR, and The Cochrane Library from the beginning of the database to June 1998 using the comprehensive search strategy recommended by the Back Review Group of the Cochrane Collaboration. INTERVENTION specific key words for this review were: patient care team, patient care management, multidisciplinary, interdisciplinary, multiprofessional, multimodal, pain clinic and functional restoration. We also reviewed reference lists and consulted the editors of the Back Review Group of the Cochrane Collaboration. SELECTION CRITERIA: DESIGN: randomised controlled trials comparing multidisciplinary bio-psycho-social rehabilitation with a non-multidisciplinary control intervention. POPULATION: Adults with disabling low back pain of more than three months in duration. INTERVENTION: Patients had to be assessed and treated by qualified professionals according to a plan that addresses physical and at least one of psychological, or social/occupational dimensions. OUTCOMES: Only trials which reported treatment effect in at least one of pain, function, employment status, quality of life or global improvement. Exclusion: Pure educational interventions (back schools) and pure physical interventions were excluded. DATA COLLECTION AND ANALYSIS: Selection, data extraction and quality grading of studies was done by two independent reviewers using pre-tested data forms. Study quality was assessed according to the scheme recommended by the Back Review Group of the Cochrane Collaboration. Trials with internal validity scores of five or more in a ten point scale were considered high quality. Discrepancies between reviewers were resolved by consensus or by a third reviewer. Given the marked heterogeneity in study settings, interventions and control groups we decided not to pool trial results in a meta-analysis. Instead, we summarized findings by strength of evidence and nature of intervention and control treatments. The evidence was judged to be strong when multiple high quality trials produced generally consistent findings. It was judged to be moderate when multiple low quality or one high quality and one or more low quality trials produced generally consistent findings. Evidence was considered to be limited when only one randomised trial existed or if findings of existing trials were inconsistent. MAIN RESULTS: Ten trials (12 randomised comparisons) were included. They randomised a total of 1964 patients with chronic low back pain. There was strong evidence that intensive multidisciplinary bio-psycho-social rehabilitation with a functional restoration approach improved function when compared with inpatient or outpatient non multidisciplinary treatments. There was moderate evidence that intensive multidisciplinary bio-psycho-social rehabilitation with a functional restoration approach improved pain when compared with outpatient non-multidisciplinary rehabilitation or usual care. There was contradictory evidence regarding vocational outcomes of intensive multidisciplinary bio-psycho-social intervention. Some trials reported improvements in work readiness, but others showed no significant reduction in sickness leaves. Less intensive outpatient psycho-physical treatments did not improve pain, function or vocational outcomes when compared with non-multidisciplinary outpatient therapy or usual care. Few trials reported effects on quality of life or global assessments. REVIEWER'S CONCLUSIONS: The reviewed trials provide evidence that intensive multidisciplinary bio-psycho-social rehabilitation with a functional restoration approach improves pain and function. Less intensive interventions did not show improvements in clinically relevant outcomes. PMID- 11869582 TI - Nutritional supplementation for stable chronic obstructive pulmonary disease. AB - BACKGROUND: Low body weight in patients with chronic obstructive pulmonary disease (COPD) is associated with an impaired pulmonary status, reduced diaphragmatic mass, lower exercise capacity and higher mortality rate when compared to adequately nourished individuals with this disease. Nutritional support may therefore be a useful part of their comprehensive care. OBJECTIVES: To conduct a systematic review of randomized controlled trials (RCTs) to clarify whether nutritional supplementation (caloric supplementation for at least 2 weeks) improved anthropometric measures, pulmonary function, respiratory muscle strength and functional exercise capacity in patients with stable COPD. SEARCH STRATEGY: Randomized controlled trials (RCTs) were identified from the Cochrane Airways Group register of RCTs, a hand-search of abstracts presented at international meetings and consultation with experts. SELECTION CRITERIA: Two reviewers independently selected trials for inclusion, assessed quality and extracted the data. DATA COLLECTION AND ANALYSIS: Within each trial and for each outcome, we calculated an effect size. The effect sizes were then pooled by a random-effects model. Homogeneity among the effect sizes was also tested. MAIN RESULTS: From 272 references, nine RCTs were ultimately included. Six papers were considered as high quality and only two studies were double-blinded. For each of the outcomes studied, the effect of nutritional support was small: the 95% confidence intervals around the pooled effect sizes all included zero. The effect of nutritional support was homogeneous across studies. An additional search conducted in August 2001 did not identify further studies. REVIEWER'S CONCLUSIONS: Nutritional support had no significant effect on anthropometric measures, lung function or exercise capacity in patients with stable COPD. PMID- 11869583 TI - Effectiveness and cost effectiveness of counselling in primary care. AB - BACKGROUND: Counsellors are prevalent in primary care settings. However, there are concerns about the clinical and cost-effectiveness of the treatments they provide, compared with alternatives such as usual care from the general practitioner, medication or other psychological therapies. OBJECTIVES: To assess the effectiveness and cost effectiveness of counselling in primary care by reviewing cost and outcome data in randomised controlled trials, controlled clinical trials and controlled patient preference trials of counselling interventions in primary care, for patients with psychological and psychosocial problems considered suitable for counselling. SEARCH STRATEGY: The original search strategy included electronic searching of databases (including the CCDAN Register of RCTs and CCTs) along with handsearching of a specialist journal. Published and unpublished sources (clinical trials, books, dissertations, agency reports etc.) were searched, and their reference lists scanned to uncover further controlled trials. Contact was made with subject experts and CCDAN members in order to uncover further trials. For the updated review, searches were restricted to those databases judged to be high yield in the first version of the review: MEDLINE, EMBASE, PSYCLIT and CINAHL, the Cochrane Controlled Trials register and the CCDAN trials register. SELECTION CRITERIA: All controlled trials comparing counselling in primary care with other treatments for patients with psychological and psychosocial problems considered suitable for counselling. Trials completed before the end of June 2001 were included in the review. DATA COLLECTION AND ANALYSIS: Data were extracted using a standardised data extraction sheet. The relevant data were entered into the Review Manager software. Trials were quality rated, using CCDAN criteria, to assess the extent to which their design and conduct were likely to have prevented systematic error. Continuous measures of outcome were combined using standardised mean differences. An overall effect size was calculated for each outcome with 95% confidence intervals. Continuous data from different measuring instruments were transformed into a standard effect size by dividing mean values by standard deviations. In view of the diversity of counselling services in primary care (the range of treatments, patients and practitioners) tests of heterogeneity were done to assess the feasibility of aggregating measures of outcome from trials. Sensitivity analyses were undertaken to test the robustness of the results. MAIN RESULTS: Seven trials were included in the review. The main analyses showed significantly greater clinical effectiveness in the counselling group compared with 'usual care' in the short term (standardised mean difference -0.28, 95% CI -0.43 to -0.13, n=772, 6 trials) but not the long-term (standardised mean difference -0.09, 95% CI -0.27 to 0.10, n=475, 4 trials). Levels of satisfaction with counselling were high. Four studies reported similar total costs associated with counselling and usual care over the long-term. However, the economic analyses were likely to be underpowered. REVIEWER'S CONCLUSIONS: Counselling is associated with modest improvement in short-term outcome compared to 'usual care', but provides no additional advantages in the long-term. Patients are satisfied with counselling, and it may not be associated with increased costs. PMID- 11869584 TI - Antidepressant and benzodiazepine for major depression. AB - BACKGROUND: Anxiety frequently coexists with depression. Adding benzodiazepines to antidepressants is commonly used to treat people with depression, although there has been no convincing evidence to show that such a combination is more effective than antidepressants alone and that there are suggestions that benzodiazepines may lose their efficacy with long-term administration and that their chronic use carries risks of dependence. OBJECTIVES: To determine whether, among adult patients with major depression, adding benzodiazepines to antidepressants brings about any benefit in terms of symptomatic recovery or side effects in the short term (less than 8 weeks) and long term (more than 2 months), in comparison with treatment by antidepressants alone. SEARCH STRATEGY: We searched MEDLINE (1972 to September 1997), EMBASE (1980 to September 1997), International Pharmaceutical Abstracts (1972 to September 1997), Biological Abstracts (1984 to September 1997), LILACS (1980 to September 1997), PsycLIT (1974 to September 1997), the Cochrane Library (issue 3, 1997) and the trial register of the Cochrane Depression, Anxiety and Neurosis Group (last searched March 1999), combined with hand searching, reference searching, SciSearch and personal contacts. SELECTION CRITERIA: All randomised controlled trials that compared combined antidepressant-benzodiazepine treatment with antidepressant alone for adult patients with major depression. Exclusion criteria are: antidepressant dosage lower than 100 mg of imipramine or its equivalent daily and duration of trial shorter than four weeks. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed the eligibility and quality of the studies. Two reviewers independently extracted the data. Standardized weighted mean differences and relative risks were estimated with random effects model. The dropouts were assigned the least favourable outcome. Two sensitivity analyses examined the effect of this assumption as well as the effect of including medium quality studies. Three a priori subgroup analyses were performed with regard to the patients with or without comorbid anxiety and with regard to the type. MAIN RESULTS: Aggregating nine studies with a total of 679 patients, the combination therapy group was less likely to drop out than the antidepressant alone group (relative risk 0.63, 95% confidence interval 0.49 to 0.81). The intention-to treat analysis (with people dropping out assigned the least favourable outcome) showed that the combination group was more likely to show improvement in their depression (defined as 50% or greater reduction in the depression scale from baseline) (relative risk 1.63, 95% confidence interval 1.18 to 2.27 at one week and relative risk 1.38, 95% confidence interval 1.15 to 1.66 at four weeks). The difference was no longer significant at six to eight weeks. None of the included RCTs lasted longer than eight weeks. The patients allocated to the combination therapy were less likely to drop out from the treatment due to side effects than those receiving antidepressants alone (relative risk 0.53, 95% confidence interval 0.32 to 0.86). However, these two groups of patients were equally likely to report at least one side effect (relative risk 0.99, 95% confidence interval 0.92 to 1.07). REVIEWER'S CONCLUSIONS: The potential benefits of adding a benzodiazepine to an antidepressant must be balanced judiciously against possible harms including development of dependence and accident proneness, on the one hand, and against continued suffering following no response and drop-out, on the other. PMID- 11869585 TI - Pygeum africanum for benign prostatic hyperplasia. AB - BACKGROUND: Benign prostatic hyperplasia (BPH), nonmalignant enlargement of the prostate, can lead to obstructive and irritative lower urinary tract symptoms (LUTS). The pharmacologic use of plants and herbs (phytotherapy) for the treatment of LUTS associated with BPH has been growing steadily. The extract of the African prune tree, Pygeum africanum, is one of the several phytotherapeutic agents available for the treatment of BPH. OBJECTIVES: To investigate the evidence whether extracts of Pygeum africanum (1) are more effective than placebo in the treatment of Benign Prostatic Hyperplasia (BPH), (2) are as effective as standard pharmacologic BPH treatments, and (3) have less side effects compared to standard BPH drugs. SEARCH STRATEGY: Trials were searched in computerized general and specialized databases (MEDLINE (1966-2000), EMBASE, Cochrane Library, Phytodok), by checking bibliographies, and by contacting relevant manufacturers and researchers. SELECTION CRITERIA: Trials were eligible if they (1) were randomized (2) included men with BPH (3) compared preparations of Pygeum africanum (alone or in combination) with placebo or other BPH medications (4) included clinical outcomes such as urologic symptom scales, symptoms, or urodynamic measurements. Eligibility was assessed by at least two independent observers. DATA COLLECTION AND ANALYSIS: Information on patients, interventions, and outcomes were extracted by at least two independent reviewers using a standard form. The main outcome measure for comparing the effectiveness of Pygeum africanum with placebo and standard BPH medications was the change in urologic symptoms scale scores. Secondary outcomes included change in urologic symptoms including nocturia and urodynamic measures (peak and mean urine flow, prostate size). The main outcome measure for adverse effects was the number of men reporting adverse effects. MAIN RESULTS: A total of 18 randomized controlled trials involving 1562 men met inclusion criteria and were analyzed. Only one of the studies reported a method of treatment allocation concealment, though 17 were double-blinded. There were no studies comparing Pygeum africanum to standard pharmacologic interventions such as alpha-adrenergic blockers or 5-alpha reductase inhibitors. The mean study duration was 64 days (range, 30-122 days). Many studies did not report results in a method that permitted meta-analysis. Compared to men receiving placebo, Pygeum africanum provided a moderately large improvement in the combined outcome of urologic symptoms and flow measures as assessed by an effect size defined by the difference of the mean change for each outcome divided by the pooled standard deviation for each outcome (-0.8 SD [95% confidence interval (CI), -1.4, -0.3 (n=6 studies)]). Men using Pygeum africanum were more than twice as likely to report an improvement in overall symptoms (RR=2.1, 95% CI = 1.4, 3.1). Nocturia was reduced by 19%, residual urine volume by 24% and peak urine flow was increased by 23%. Adverse effects due to Pygeum Africanum were mild and comparable to placebo. The overall dropout rate was 12% and was similar between Pygeum Africanum (13%), placebo (11%) and other controls (8%). REVIEWER'S CONCLUSIONS: A standardized preparation of Pygeum africanum may be a useful treatment option for men with lower urinary symptoms consistent with benign prostatic hyperplasia. However, the reviewed studies were small in size, were of short duration, used varied doses and preparations and rarely reported outcomes using standardized validated measures of efficacy. Additional placebo controlled trials are needed as well as studies that compare Pygeum africanum to active controls that have been convincingly demonstrated to have beneficial effects on lower urinary tract symptoms related to BPH. These trials should be of sufficient size and duration to detect important differences in clinically relevant endpoints and use standardized urologic symptom scale scores. PMID- 11869586 TI - Therapeutic hypothermia for head injury. AB - BACKGROUND: Induced hypothermia has been used in the treatment of head injury for many years. Encouraging results from small trials and laboratory studies led to renewed interest in the area and some larger trials. OBJECTIVES: To estimate the effects of mild induced hypothermia in moderate and severe head injury on mortality, long-term functional outcome, complications, and short-term control of intracranial pressure (ICP). SEARCH STRATEGY: We searched the Injuries Group Specialised register (last searched in 2001), Medline, EMBASE and the Cochrane Controlled Trials Register. We handsearched conference proceedings and checked reference lists of relevant articles. SELECTION CRITERIA: Randomised controlled trials of mild hypothermia to 34-35 Celsius for a t least 12 hours versus control (open or normothermia) in patients with any closed head injury requiring hospitalisation. Two reviewers independently assessed all trials. DATA COLLECTION AND ANALYSIS: Data on death, Glasgow Outcome Scale, complications and ICP were sought and extracted, either from published material or by contacting the investigators. Odds ratios and 95% confidence intervals were calculated for each trial on an intention-to-treat basis. Quantitative synthesis of data on complications other than pneumonia or ICP was not attempted. Trials of immediate and deferred hypothermia were analysed separately. MAIN RESULTS: We found 12 trials with 812 participants. Active immediate hypothermia was associated with an odds ratio for death of 0.88, (771 patients, OR 0.88, 95% CI 0.63 to 1.21), and 0.75 for odds of being dead or severely disabled, (746 patients, OR 0.75, 95% CI 0.56 to 1.00). Hypothermia treatment was associated with a statistically significant increase in odds of pneumonia (281 patients, OR 1.95, 95% CI 1.18 to 3.23). The trial of deferred hypothermia (33 patients) reported a huge but not statistically significant reduction in the odds of death at 6 months, (OR 0.21, 95% CI 0.04 to 1.05). For death or severe disability deferred hypothermia was associated with an odds ratio of 0.10 (95% CI 0.01 to 1.00). REVIEWER'S CONCLUSIONS: There is no evidence that hypothermia is beneficial in the treatment of head injury. The earlier, encouraging, trial results have not been repeated in larger trials. The reasons for this are unclear. Hypothermia increases the risk of pneumonia and has other potentially harmful side effects. Therefore, it would seem inappropriate to use this intervention outside of controlled trials in subgroups of patients for whom there is good reason to think the treatment would be beneficial. PMID- 11869587 TI - Calcium supplementation during pregnancy for preventing hypertensive disorders and related problems. AB - BACKGROUND: Calcium supplementation may prevent high blood pressure through a number of mechanisms and may help to prevent preterm labour. OBJECTIVES: The objective of this review was to assess the effects of calcium supplementation during pregnancy on hypertensive disorders of pregnancy and related maternal and child adverse outcomes. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register (October 2001) and the Cochrane Controlled Trials Register (Issue 3, 2001) and we contacted study authors. SELECTION CRITERIA: Randomised trials comparing at least one gram daily of calcium during pregnancy with placebo. DATA COLLECTION AND ANALYSIS: Eligibility and trial quality were assessed. Data extraction was carried out and double entered. MAIN RESULTS: Eleven studies were included, all of good quality. There was a modest reduction in high blood pressure with calcium supplementation (relative risk 0.81, 95% confidence interval 0.74 to 0.89). The effect was greatest for women at high risk of hypertension (relative risk 0.45, 95% confidence interval 0.31 to 0.66) and those with low baseline dietary calcium (relative risk 0.49, 95% confidence interval 0.38 to 0.62). There was also a modest reduction in the risk of pre-eclampsia with calcium supplementation (relative risk 0.68, 95% confidence interval 0.57 to 0.81). The effect was greatest for women at high risk of hypertension (relative risk 0.21, 95% confidence interval 0.11 to 0.39) and those with low baseline calcium intake (relative risk 0.32, 95% confidence interval 0.21 to 0.49). There was no overall effect on the risk of preterm delivery, although there was a reduction in risk amongst women at high risk of hypertension (relative risk 0.42, 95% confidence interval 0.23 to 0.78). There was no evidence of any effect of calcium supplementation on stillbirth or death before discharge from hospital. There were fewer babies with birthweight < 2500g (RR 0.83, 95% CI 0.71-0.98). In one study, childhood systolic blood pressure > 95th percentile was reduced (RR 0.59, 95% CI 0.39-0.91). REVIEWER'S CONCLUSIONS: Calcium supplementation appears to be beneficial for women at high risk of gestational hypertension and in communities with low dietary calcium intake. Optimum dosage requires further investigation. PMID- 11869588 TI - Piracetam for fetal distress in labour. AB - BACKGROUND: Piracetam is thought to promote the metabolism of brain cells when they are hypoxic. It has been used to prevent adverse effects of fetal distress. OBJECTIVES: The objective of this review was to assess the effects of piracetam for suspected fetal distress in labour on method of delivery and perinatal morbidity. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register (The Cochrane Library Issue 3, 2001) were searched. Date of last search: September 2001. SELECTION CRITERIA: Randomised trials of piracetam compared with placebo or no treatment for suspected fetal distress in labour. DATA COLLECTION AND ANALYSIS: Both reviewers assessed eligibility and trial quality. MAIN RESULTS: One study of 96 women was included. Piracetam compared with placebo was associated with a trend to reduced need for caesarean section (relative risk 0.57, 95% confidence interval 0.32 to 1.03). There were no statistically significant differences in relative risk between the piracetam and placebo group for neonatal morbidity (measured by neonatal respiratory distress) or Apgar score. REVIEWER'S CONCLUSIONS: There is not enough evidence to evaluate the use of piracetam for fetal distress in labour. PMID- 11869589 TI - Antibiotic regimens for endometritis after delivery. AB - BACKGROUND: Post-partum endometritis, which is more common after cesarean section, occurs when vaginal organisms invade the endometrial cavity during labour and birth. Antibiotic treatment is warranted. OBJECTIVES: The effect of different antibiotic regimens for the treatment of postpartum endometritis on failure of therapy and complications was systematically reviewed. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group's trials register and the Cochrane Controlled Trials Register. Date of last search: June 2001. SELECTION CRITERIA: Randomised trials of different antibiotic regimens for postpartum endometritis, after cesarean section or vaginal birth, where outcomes of treatment failure or complications were reported were selected. DATA COLLECTION AND ANALYSIS: Data were abstracted independently by the reviewers. Comparisons were made between different types of antibiotic regimen, based on type of antibiotic and duration and route of administration. Summary relative risks were calculated. MAIN RESULTS: Forty-seven trials were included. Overall the studies were methodologically poor. In the intent-to-treat analysis, fifteen studies comparing clindamycin and an aminoglycoside with another regimen showed more treatment failures with another regimen (relative risk (RR) 1.32; 95% confidence interval (CI) 1.09-1.60). Failures of those regimens with poor activity against penicillin resistant anaerobic bacteria were more likely (RR 1.53; 95% CI 1.10-2.13). In four studies that compared continued oral antibiotic therapy after intravenous therapy, no differences were found in recurrent endometritis or other outcomes. There was no evidence of difference in incidence of allergic reactions. Cephalosporins were associated with less diarrhea. REVIEWER'S CONCLUSIONS: The combination of gentamicin and clindamycin is appropriate for the treatment of endometritis. Regimens with activity against penicillin resistant anaerobic bacteria are better than those without. There is no evidence that any one regimen is associated with fewer side effects. Once uncomplicated endometritis has clinically improved with intravenous therapy, oral therapy is not needed. PMID- 11869590 TI - Servo-control for maintaining abdominal skin temperature at 36C in low birth weight infants. AB - BACKGROUND: Randomized trials have shown that the neonatal mortality rate of low birth-weight babies can be reduced by keeping them warm. For low birth-weight babies nursed in incubators, warm conditions may be achieved either by heating the air to a desired temperature, or by servo-controlling the baby's body temperature at a desired set-point. OBJECTIVES: In low birth weight infants, to determine the effect on death and other important clinical outcomes of targeting body temperature rather than air temperature as the end-point of control of incubator heating. SEARCH STRATEGY: Standard search strategy of the Cochrane Neonatal Review Group. Searches were made of the Cochrane Controlled Trials Register (CCTR) (Cochrane Library, Issue 4, 2001) and MEDLINE, 1966 to November 2001. SELECTION CRITERIA: Randomized or quasi-randomized trials which test the effects of having the heat output of the incubator servo-controlled from body temperature compared with setting a constant incubator air temperature. DATA COLLECTION AND ANALYSIS: Trial methodologic quality was systematically assessed. Outcome measures included death, timing of death, cause of death, and other clinical outcomes. Categorical outcomes were analyzed using relative risk and risk difference. Meta-analysis assumed a fixed effect model. MAIN RESULTS: Two eligible trials were found. In total, they included 283 babies and 112 deaths. Compared to setting a constant incubator air temperature of 31.8C, servo-control of abdominal skin temperature at 36C reduces the neonatal death rate among low birth weight infants: relative risk 0.72 (95% CI 0.54, 0.97); risk difference 12.7% (95% CI -1.6, -23.9). This effect is even greater among VLBW infants. REVIEWER'S CONCLUSIONS: During at least the first week after birth, low birth weight babies should be provided with a carefully regulated thermal environment that is near the thermoneutral point. For LBW babies in incubators, this can be achieved by adjusting incubator temperature to maintain an anterior abdominal skin temperature of at least 36C, using either servo-control or frequent manual adjustment of incubator air temperature. PMID- 11869591 TI - Intravenous immunoglobulin for treating sepsis and septic shock. AB - BACKGROUND: Death from severe sepsis and septic shock is common, and researchers have explored whether antibodies to the endotoxins in some bacteria reduces mortality. OBJECTIVES: To estimate the effects of intravenous immunoglobulin (IVIG) in patients with bacterial sepsis or septic shock on mortality, bacteriological failure rates, and duration of stay in hospital. SEARCH STRATEGY: We searched the Cochrane Infectious Diseases Group specialized register up to November 2001; the Cochrane Controlled Trials Register, The Cochrane Library issue 4, 2001; MEDLINE 1966 to November 2001; and EMBASE 1988 to September 2001. We contacted investigators active in the field for unpublished data. SELECTION CRITERIA: Randomised trials comparing intravenous immunoglobulin (monoclonal or polyclonal) with placebo or no intervention, in patients with bacterial sepsis or septic shock. DATA COLLECTION AND ANALYSIS: Inclusion criteria, trial quality assessment, and data abstraction were done in duplicate. We conducted pre specified subgroup analyses by type of immunoglobulin preparation. MAIN RESULTS: Twenty-seven out of 55 studies met our inclusion criteria. Pooled analysis of all types of IVIG preparations revealed a significant trend toward reduction of mortality (n= 8,856; RR=0.91; 95%CI 0.86-0.96). Overall mortality was reduced in patients who received polyclonal IVIG (n=492; RR=0.64; 95% CI 0.51 to 0.80). For the two high-quality trials on polyclonal IVIG, the RR for overall mortality was 0.30, but the confidence interval was wide (95% CI 0.09 to 0.99, n=91). Mortality was not reduced among patients who received monoclonal antibodies such as anti endotoxins (n=2,826 in 5 good-quality studies; RR=0.97; 95% CI 0.88 to 1.07) or anti-cytokines (n=4,318; RR=0.93; 95% CI 0.86 to 1.01). A few studies measured secondary outcomes (deaths from sepsis or length of hospitalisation) but no differences in the intervention and control groups were identified except among those who received polyclonal IVIG, where sepsis-related mortality was significantly reduced (n=161; RR=0.35; 95% CI 0.18 to 0.69). REVIEWER'S CONCLUSIONS: Polyclonal IVIG significantly reduced mortality and and is a promising adjuvant in the treatment of sepsis and septic shock. However, all the trials were small and the totality of the evidence is insufficient to support a robust conclusion of benefit. Adjunctive therapy with monoclonal IVIGs remains experimental. PMID- 11869592 TI - Interventions for preventing and treating pelvic and back pain in pregnancy. AB - BACKGROUND: More than a third of women experience back pain during pregnancy. The pain can interfere with work, daily activities and sleep. OBJECTIVES: The objective of the review was to assess the effects of preventive interventions and treatments for pelvic and back pain in pregnancy. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register (October 2001) and the Cochrane Controlled Trials Register (The Cochrane Library, Issue 3, 2001). SELECTION CRITERIA: Randomised trials of any treatment to reduce the incidence or severity of pelvic/back pain in pregnancy, or to prevent pelvic/back pain arising in pregnancy. DATA COLLECTION AND ANALYSIS: Trial quality was assessed and data were extracted independently by two reviewers. MAIN RESULTS: Three trials are included in this review involving 376 women. One randomized trial compared water gymnastics from 20 weeks with no treatment. The authors report less pain in the treatment group but the data are hard to interpret; there was a difference in rates of absence from work after 32 weeks of pregnancy (odds ratio 0.38, 95% confidence intervals 0.16-0.88). In another trial, acupuncture was rated as giving 'good' or 'excellent' help more frequently than physiotherapy (odds ratio 6.58, 95% confidence intervals 1.0-43.16) but this may reflect the benefit of individual compared with group therapy. One trial of 109 women compared the use of a special shaped pillow to fit under the woman's abdomen (Ozzlo pillow) with a standard pillow. Fewer women rated the Ozzlo pillow of 'little help' compared with the standard pillow (odds ratio 0.32, 95% confidence interval 0.18 to 0.58). REVIEWER'S CONCLUSIONS: Water gymnastics appear to reduce back pain in pregnancy. More women are able to continue at work. Specially shaped pillows help reduce back pain in late pregnancy and improve sleep. It is a pity that the Ozzlo pillow seems no longer to be available. Both physiotherapy and acupuncture may reduce back and pelvic pain. Individual acupuncture sessions were rated as more help than group physiotherapy sessions. PMID- 11869593 TI - Support for breastfeeding mothers. AB - BACKGROUND: Both observational and recent experimental evidence support the promotion of breastfeeding as the optimal form of infant nutrition. There is, however, uncertainty as to the most effective way of providing support to women who choose to breastfeed their children. A systematic review was performed to describe studies undertaken in this area and to assess the effectiveness of supplementary support. OBJECTIVES: The objective of this review was to assess the effects of breastfeeding support. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register, the Cochrane Controlled Trials Register, MEDLINE and EMBASE. These were last searched in March 2001. Secondary references were searched and researchers in the field were contacted. SELECTION CRITERIA: Controlled trials of acceptable quality comparing extra support for breastfeeding mothers with usual maternity care. DATA COLLECTION AND ANALYSIS: Data were extracted by one reviewer and checked by a second reviewer. MAIN RESULTS: Twenty eligible randomised or quasi-randomised controlled trials from 10 countries were identified involving 23,712 mother-infant pairs. There was a beneficial effect on the duration of any breastfeeding in the meta-analysis of all forms of extra support (relative risk (RR) for stopping any breastfeeding before six months 0.88 [95% confidence interval (CI) 0.81,0.95]; 15 trials, 21,910 women). The effect was greater for exclusive breastfeeding (RR 0.78 [95% CI 0.69,0.89]; 11 trials, 20,788 women). Extra professional support appeared beneficial for any breastfeeding (RR 0.89 [95% CI 0.81,0.97]; 10 trials, 19,696 women) and for exclusive breastfeeding (RR 0.90 [95% confidence interval 0.81,1.01]; six trials, 18,258 women) although the latter effect did not achieve full statistical significance. Lay support was effective in reducing the cessation of exclusive breastfeeding (RR 0.66 [95% CI 0.49,0.89]; five trials, 2530 women) but its effect on any breastfeeding did not reach statistical significance (RR 0.84 [95% CI 0.69,1.02]; five trials, 2224 women). Professional support in the largest trial to assess health outcomes produced a significant reduction in the risk of gastro-intestinal infections and atopic eczema. In two trials with children suffering from diarrhoeal illness extra support was highly effective in increasing short term exclusive breastfeeding rates and reducing recurrence of diarrhoea. REVIEWER'S CONCLUSIONS: Consideration should be given to providing supplementary breastfeeding support as part of routine health service provision. There is clear evidence for the effectiveness of professional support on the duration of any breastfeeding although the strength of its effect on the rate of exclusive breastfeeding is uncertain. Lay support is effective in promoting exclusive breastfeeding while the strength of its effect on the duration of any breastfeeding is also uncertain. Evidence supports the promotion of exclusive breastfeeding as central to the management of diarrhoeal illness in partially breastfed infants. Further trials are required to assess the effectiveness (including cost-effectiveness) of both lay and professional support in different settings - in particular in those communities with low rates of breastfeeding initiation. Research is also required into the most appropriate training for those, whether lay or professional, who support breastfeeding mothers. PMID- 11869594 TI - Nerve blocks (subcostal, lateral cutaneous, femoral, triple, psoas) for hip fractures. AB - BACKGROUND: Various nerve blocks using local anaesthetic agents have been used in order to reduce pain after hip fracture. OBJECTIVES: To determine the effects of nerve blocks (inserted either pre-operatively, operatively or post-operatively) as part of the treatment for a hip fracture. SEARCH STRATEGY: We searched the Cochrane Musculoskeletal Injuries Group specialised register (October 2001), MEDLINE -OVID WEB (1996 to October 2001) and reference lists of relevant articles. SELECTION CRITERIA: Randomised and quasi-randomised trials involving the use of nerve blocks as part of the care of a hip fracture patient. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed trial quality, by use of a nine item scale, and extracted data. Wherever appropriate, results of outcome measures were pooled. MAIN RESULTS: Eight randomised or quasi-randomised trials involving 328 patients were included. Three trials related to insertion of a nerve block pre-operatively and the remaining five to peri-operative insertion. Nerve blocks resulted in a reduction of the quantity of parenteral or oral analgesia administered to control pain from the fracture/operation or during surgery and/or a reduction in reported pain levels. It was not possible to demonstrate if this reduction in analgesia use was associated with any other clinical benefit. REVIEWER'S CONCLUSIONS: Because of the small number of patients included in this review and the differing type of nerve blocks and timing of insertion, it is not possible to determine if nerve blocks confer any significant benefit when compared with other analgesic methods as part of the treatment of a hip fracture. Further trials with larger numbers of patients and full reporting of clinical outcomes would be justified. PMID- 11869595 TI - Nedocromil sodium for preventing exercise-induced bronchoconstriction. AB - BACKGROUND: Exercise-induced bronchoconstriction (EIB) following strenuous physical exertion afflicts many people. It can be the cause of sub-optimal performance, symptoms such as cough, dyspnea, wheeze and chest tightness, and can lead people to avoid physical activity. Management of EIB focuses on prevention through pharmaco-therapy and alternate strategies. Single use, pre-exercise, beta agonists and non-steroidal antiinflammatory agents are recommended. OBJECTIVES: Bronchodilator medications have been commonly used to prevent narrowing of airways after exercise, but anti-inflammatory drugs such as nedocromil sodium have also been used. The objective of this review was to assess the effects of a single dose of nedocromil sodium to prevent exercise-induced bronchoconstriction. SEARCH STRATEGY: We searched the Cochrane Airways Group trials register, the Cochrane Controlled Trials Register, Current Contents, review articles, textbooks and reference lists of articles. We also contacted the drug manufacturer and primary authors for additional citations. SELECTION CRITERIA: Randomised trials comparing a single dose of nedocromil sodium with placebo to prevent exercise induced bronchoconstriction in patients with EIB over six years of age. DATA COLLECTION AND ANALYSIS: Trial quality assessment and data extraction were conducted independently by two reviewers. Study authors were contacted for confirmation of data. MAIN RESULTS: The combined results from 20 randomised controlled trials involving 280 participants, show that 4 mg, of nedocromil sodium inhaled 15 to 60 minutes prior to exercise significantly reduce the severity and duration of EIB in both adults and children, when compared to placebo. The maximum percentage fall in FEV1 was improved significantly compared to placebo (weighted mean difference 15.5 %; 95% confidence interval:13.2 to 18.1). For the maximum percentage fall in peak expiratory flow rate (PEFR) the improvement was similar: WMD 15.0%, (95% CI 8.3 to 21.6). Nedocromil shortened the time to recover lung normal function from more than 30 minutes with placebo to less than 10 minutes with the drug. It had a greater effect on those patients with more severe exercise-induced bronchoconstriction (defined as an exercise induced fall in lung function > 30% from baseline). There were no significant adverse effects reported with the short term use of nedocromil. A further search conducted in September 2001 did not yield any further studies. REVIEWER'S CONCLUSIONS: Nedocromil sodium used before exercise reduces the severity and duration of exercise-induced bronchoconstriction. This effect appears to be more pronounced in people with severe exercise-induced bronchoconstriction. PMID- 11869596 TI - Human albumin solution for resuscitation and volume expansion in critically ill patients. AB - BACKGROUND: Human albumin solutions are used in a range of medical and surgical problems. Licensed indications are the emergency treatment of shock and other conditions where restoration of blood volume is urgent, burns, and hypoproteinaemia. Human albumin solutions are more expensive than other colloids and crystalloids. OBJECTIVES: To quantify the effect on mortality of human albumin and plasma protein fraction (PPF) administration in the management of critically ill patients. SEARCH STRATEGY: We searched the Cochrane Injuries Group trials register, Cochrane Controlled Trials Register, Medline, Embase and BIDS Index to Scientific and Technical Proceedings. Reference lists of trials and review articles were checked, and authors of identified trials were contacted. The search was last updated in November 2001. SELECTION CRITERIA: Randomised controlled trials comparing albumin/PPF with no albumin/PPF, or with a crystalloid solution, in critically ill patients with hypovolaemia, burns or hypoalbuminaemia. DATA COLLECTION AND ANALYSIS: We collected data on the participants, albumin solution used, mortality at the end of follow up, and quality of allocation concealment. Analysis was stratified according to patient type. MAIN RESULTS: We found 31 trials meeting the inclusion criteria and reporting death as an outcome. There were 177 deaths among 1519 trial participants. For each patient category the risk of death in the albumin treated group was higher than in the comparison group. For hypovolaemia the relative risk of death following albumin administration was 1.46 (95% confidence interval 0.97 to 2.22), for burns the relative risk was 2.40 (1.11 to 5.19), and for hypoalbuminaemia the relative risk was 1.38 (0.94 to 2.03). The pooled relative risk of death with albumin administration was 1.52 (1.17 to 1.99). Overall, the risk of death in patients receiving albumin was 14% compared to 9% in the control groups, an increase in the risk of death of 5% (2% to 8%). These data suggest that for every 20 critically ill patients treated with albumin there is one additional death. REVIEWER'S CONCLUSIONS: There is no evidence that albumin administration reduces the risk of death in critically ill patients with hypovolaemia, burns or hypoalbuminaemia, and a strong suggestion that it may increase the risk of death. These data suggest that the use of human albumin in critically ill patients should be urgently reviewed and that it should not be used outside the context of a rigorously conducted randomised controlled trial. PMID- 11869597 TI - Ultrasound therapy for acute ankle sprains. AB - BACKGROUND: Ultrasound is used in the treatment of a wide variety of musculoskeletal disorders. OBJECTIVES: To evaluate the effects of ultrasound therapy in the treatment of acute ankle sprains. SEARCH STRATEGY: We searched the Cochrane Musculoskeletal Injuries Group specialised register (November 15th 2001), the Cochrane Controlled Trials Register (The Cochrane Library Issue 3, 2001), MEDLINE (1966 to October 2001), EMBASE (1983 to 2001 week 45), CINAHL (1982 to October week 4 2001), and PEDro - The Physiotherapy Evidence Database (http://ptwww.cchs.usyd.edu.au/pedro/ accessed 15.11.01). We also searched the Cochrane Rehabilitation and Related Therapies Field database, reference lists of articles, and contacted colleagues. SELECTION CRITERIA: Randomised or pseudo randomised trials were included in the review if the following conditions were met: at least one study group was treated with active ultrasound; the study included patients with acute lateral ankle sprains; and outcome measures included general improvement, pain, swelling, functional disability, or range of motion. Final selection of papers was conducted by two independent reviewers. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed trial quality using a standardised checklist, and extracted data. Whenever possible, the results were analysed on an intention-to-treat basis. Relative risks together with 95 per cent confidence intervals were calculated for dichotomous outcomes, and weighted or, where different scales were used, standardised mean differences together with 95 per cent confidence intervals for continuous outcome measures. Pooling of data was undertaken where there was clinical homogeneity in terms of subjects, treatments, outcomes, and follow-up time points. MAIN RESULTS: Five trials were included, involving 572 patients. Four of these trials were only of modest methodological quality and one placebo-controlled trial was considered to be of good quality. None of the four placebo-controlled trials (sham ultrasound) could demonstrate statistically significant differences between true and sham ultrasound therapy for any outcome measure at seven to 14 days of follow-up. The pooled relative risk for general improvement was 1.04 (random effects model, 95% confidence interval 0.92 to 1.17) for the comparison between ultrasound and sham ultrasound. The differences between intervention groups were generally small, between zero and six per cent for most dichotomous outcomes. However, one trial reported relatively large differences for pain free status (20%) and swelling (25%) in favour of ultrasound treatment. REVIEWER'S CONCLUSIONS: The extent and quality of the available evidence for the effects of ultrasound therapy for acute ankle sprains is limited. The results of four placebo-controlled trials do not support the use of ultrasound in the treatment of ankle sprains. The magnitude of most reported treatment effects appeared to be small, and may be of limited clinical importance. As yet, only few trials are available and no conclusions can be made regarding an optimal and adequate dosage schedule for ultrasound therapy, and whether such a schedule would improve on the reported effectiveness of ultrasound for ankle sprains. PMID- 11869598 TI - Anticholinergic drugs for wheeze in children under the age of two years. AB - BACKGROUND: Wheeze in infancy and early childhood is common and appears to be increasing though the magnitude of any increase is unclear. Most wheezing episodes in infancy are precipitated by respiratory viral infections. Treatment of very young children with wheeze remains controversial. Anti-cholinergics are often prescribed but practice varies widely and the efficacy of this form of therapy remains the subject for debate. OBJECTIVES: Wheeze in infancy and early childhood is common and appears to be increasing. Most wheezing episodes in infancy are a result of viral infection. Bronchodilator medications such as beta2 agonists and anti-cholinergic agents are often used to relieve symptoms, but patterns of use vary. The objective of this review was to assess the effects of anti-cholinergic therapy in the treatment of wheezing infants. SEARCH STRATEGY: We searched the Cochrane Airways Group trials register and the reference lists of articles. We contacted researchers in the field and industry sources. SELECTION CRITERIA: Randomised trials that compared anti-cholinergic therapy with placebo or beta2-agonists in wheezing children under two years of age. Children with acute bronchiolitis and chronic lung disease were excluded. DATA COLLECTION AND ANALYSIS: Eligibility for inclusion and quality of trials were assessed independently by two reviewers. MAIN RESULTS: Six trials involving 321 infants in three different settings were included. Compared with beta2-agonist alone, the combination of ipratropium bromide and beta2-agonist was associated with a reduced need for additional treatment, but no difference was seen in treatment response, respiratory rate or oxygen saturation improvement in the emergency department. There was no significant difference in length of hospital stay between ipratropium bromide and placebo; or between ipratropium bromide and beta2 agonist combined compared with beta2-agonist alone. However, combined ipratropium bromide and beta2-agonist compared to placebo showed significantly improved clinical scores at 24 hours. Parents preferred ipratropium bromide over nebulised water or placebo for relief of their children's symptoms at home. REVIEWER'S CONCLUSIONS: There is not enough evidence to support the uncritical use of anti cholinergic therapy for wheezing infants, although parents using it at home were able to identify benefits. PMID- 11869599 TI - Extracorporeal membrane oxygenation for severe respiratory failure in newborn infants. AB - BACKGROUND: Extracorporeal membrane oxygenation (ECMO) is a complex procedure of life support in severe but potentially reversible respiratory failure, used particularly in mature newborn infants. Although the number of babies requiring ECMO is small, and the ECMO policy invasive and potentially expensive, its benefits may be high. OBJECTIVES: To determine whether ECMO used for neonatal infants with severe respiratory failure is clinically effective and cost effective compared to a policy of conventional ventilatory support. SEARCH STRATEGY: The Cochrane Neonatal Group Specialised Register, the Cochrane Controlled Trials Register, and MEDLINE were searched for 1974 to 2001. SELECTION CRITERIA: All randomised trials comparing neonatal ECMO to conventional ventilatory support. DATA COLLECTION AND ANALYSIS: The authors independently evaluated the trials for methodological quality and appropriateness for inclusion in the Review (without consideration of their results), and then independently extracted the data. MAIN RESULTS: The three trials from the USA and one from the UK recruited clinically similar groups of babies. Two trials excluded infants with congenital diaphragmatic hernias. In two, transfer for ECMO implied transport over a considerable distance. One study included an economic evaluation. Two trials had follow up information. All except the UK trial had very small numbers of patients. Two of the trials used conventional randomisation with low potential for bias. The other two used less usual designs which have led to difficulties in their interpretation. All four trials showed a strong benefit of ECMO on mortality (RR 0.44; 95% CI 0.31 to 0.61), especially for babies without congenital diaphragmatic hernia (RR 0.33, 95% CI 0.21 to 0.53). Only the UK trial provided information about death or disability at one and four years, and showed benefit of ECMO at one year (RR 0.56, 95% CI 0.40 to 0.78), and at four years (RR 0.62, 95% CI 0.45 to 0.86). Overall nearly half of the children had died or were severely disabled at four years of age, reflecting the severity of their underlying conditions. Based on economic analysis from the UK trial, the ECMO policy is as cost-effective as other intensive care technologies in common use. REVIEWER'S CONCLUSIONS: A policy of using ECMO in mature infants with severe but potentially reversible respiratory failure would result in significantly improved survival without increased risk of severe disability amongst survivors. For babies with diaphragmatic hernia ECMO offers short term benefits but the overall effect of employing ECMO in this group is not clear. Further studies are needed to refine ECMO techniques; to consider the optimal timing for introducing ECMO; to identify which infants are most likely to benefit; and to address the longer term implications of neonatal ECMO during later childhood and adult life. PMID- 11869600 TI - Intravenous or enteral loop diuretics for preterm infants with (or developing) chronic lung disease. AB - BACKGROUND: Lung disease in preterm infants is often complicated with lung edema. OBJECTIVES: The aim of this review was to assess the risks and benefits of administration of a diuretic acting on the loop of Henle (loop diuretic) in preterm infants with or developing chronic lung disease (CLD). Primary objectives were to assess changes in need for oxygen or ventilatory support and effects on long-term outcome, and secondary objectives were to assess changes in pulmonary mechanics and potential complications of therapy. SEARCH STRATEGY: We used the standard search method of the Cochrane Neonatal Review Group. We searched MEDLINE (1966-October 2001), EMBASE (1974-November 2001) and the Cochrane Controlled Trials Register (CCTR) (Cochrane Library, Issue 4, 2001). In addition, we hand searched several abstract books of national and international American and European Societies. SELECTION CRITERIA: We included in this analysis trials in which preterm infants with or developing chronic lung disease and at least 5 days of age were all randomly allocated to receive a loop diuretic either enterally or intravenously. Eligible studies needed to assess at least one of the outcome variables defined a priori for this systematic review. Primary outcome variables included important clinical outcomes, and secondary outcome variables included toxicity and pulmonary mechanics (e.g., lung compliance and airway resistance). DATA COLLECTION AND ANALYSIS: We used the standard method for the Cochrane Collaboration which is described in the Cochrane Collaboration Handbook. Two investigators extracted, assessed and coded separately all data for each study, using a form that was designed specifically for this review. Any disagreement was resolved by discussion. We combined parallel and cross-over trials and, whenever possible, transformed baseline and final outcome data measured on a continuous scale into change scores using Follmann's formula. MAIN RESULTS: The only loop diuretic used in the studies which met the selection criteria was furosemide. Most studies focused on pathophysiological parameters and did not assess effects on important clinical outcomes defined in this review, or the potential complications of diuretic therapy. In preterm infants < 3 weeks of age developing CLD, furosemide administration has either inconsistent effects or no detectable effect. In infants > 3 weeks of age with CLD, a single intravenous dose of 1 mg/kg of furosemide improves lung compliance and airway resistance for 1 hour. Chronic administration of furosemide improves both oxygenation and lung compliance. REVIEWER'S CONCLUSIONS: In preterm infants > 3 weeks of age with CLD, acute and chronic administration of furosemide improve lung compliance. Chronic administration of intravenous or enteral furosemide improves oxygenation. In view of the lack of data from randomized trials concerning effects on important clinical outcomes, routine or sustained use of systemic loop diuretics in infants with (or developing) CLD cannot be recommended based on current evidence. Randomized trials are needed to assess the effects of furosemide administration on survival, duration of ventilatory support and oxygen administration, length of hospital stay, potential complications and long-term outcome. PMID- 11869601 TI - Vitamin A for treating measles in children. AB - BACKGROUND: Measles is a leading cause of childhood morbidity and mortality. Vitamin A deficiency is a recognised risk factor for severe measles. The World Health Organization (WHO) recommends administration of an oral dose of 200,000 IU (or 100,000 IU in infants) of vitamin A per day for two days to children with measles in areas where vitamin A deficiency may be present. OBJECTIVES: The purpose of this review is to determine whether vitamin A when commenced after measles has been diagnosed, is beneficial in preventing mortality, pneumonia and other complications in children. SEARCH STRATEGY: MEDLINE and the Cochrane Library, Issue 4, 1999 were searched. SELECTION CRITERIA: Only randomized controlled trials in which children with measles were given vitamin A or placebo along with standard treatment were considered. DATA COLLECTION AND ANALYSIS: Studies were assessed independently by two reviewers. The analysis of dichotomous outcomes was done using the StatXact software package. Sub-group analyses were done for dose, formulation, age, hospitalisation and pneumonia specific mortality. Weighted mean difference with 95% CI were calculated for continuous outcomes. MAIN RESULTS: The relative risks (RR) and 95% Confidence Intervals (CI) are based on the estimates from the StatXact software package. There was no significant reduction in mortality in the vitamin A group when all the studies were pooled together (RR 0.60; 95% CI 0.32 to 1.12)(StatXact estimate). There was a 64% reduction in the risk of mortality in children who were given two doses of 200,000 IU of vitamin A (RR=0.36; 95% CI 0.14 to 0.82) as compared to placebo. Two doses of water based vitamin A were associated with a 81% reduction in risk of mortality (RR=0.19; 95% CI 0.02 to 0.85) as compared to 48% seen in two doses of oil based preparation (RR=0.52; 95% CI 0.16 to 1.40). Two doses of oil and water based vitamin A were associated with a 82% reduction in the risk of mortality in children under the age of 2 years (RR=0.18; 95% CI 0.03 to 0.61) and a 67% reduction in the risk of pneumonia specific mortality (RR=0.33; 95% CI 0.08 to 0.92). There was no evidence that vitamin A in a single dose of 200,000 IU was associated with a reduced risk of mortality among children with measles (RR=0.77; 95% CI 0.34 to 1.78). Sub-groups like age, dose, formulation, hospitalisation and case fatality in the study area were highly correlated and there were not enough studies to separate out the individual effects of these factors. There was a 47% reduction in the incidence of croup (RR=0.53; 95% CI 0.29 to 0.89), while there was no significant reduction in the incidence of pneumonia (RR=0.92; 95% CI 0.69 to 1.22) or of diarrhoea (RR=0.80; 95% CI 0.27 to 2.34). Duration of diarrhoea was measured in days and there was a reduction in its duration of almost two days WMD -1.92, 95% CI -3.40 to -0.44. Only one study evaluated otitis media and found a 74% reduction in its incidence (RR=0.26, 95% CI, 0.05 to 0.92). We did not find evidence that a single dose of 200,000 IU of vitamin A per day, given in oil based formulation in areas with low case fatality, was associated with reduced mortality among children with measles. However, there was evidence that the same dose given for two days was associated with a reduced risk of overall mortality and pneumonia specific mortality. REVIEWER'S CONCLUSIONS: Although we did not find evidence that a single dose of 200,000 IU of vitamin A per day was associated with reduced mortality among children with measles, there was evidence that the same dose given for two days was associated with a reduced risk of overall mortality and pneumonia specific mortality. The effect was greater in children under the age of two years. There were no trials that compared a single dose with two doses, although the precision of the estimates of trials that used a single dose were similar to the trials that used two doses. PMID- 11869602 TI - Interventions for preventing tobacco sales to minors. AB - BACKGROUND: Laws restricting sales of tobacco products to minors exist in many countries, but young people may still purchase cigarettes easily. OBJECTIVES: The review assesses the effects of interventions to reduce underage access to tobacco by deterring shopkeepers from making illegal sales. SEARCH STRATEGY: We searched the Cochrane Tobacco Addiction group trials register, MEDLINE and EMBASE. Date of the most recent searches: October 2001. SELECTION CRITERIA: We included controlled trials and uncontrolled studies with pre- and post intervention assessment of interventions to change retailers' behaviour. The outcomes were changes in retailer compliance with legislation (assessed by test purchasing), changes in young people's smoking behaviour, and perceived ease of access to tobacco products. DATA COLLECTION AND ANALYSIS: Studies were prescreened for relevance by one person and assessed for inclusion by two people independently. Data from included studies were extracted by one person and checked by a second. Study designs and types of intervention were heterogeneous so results were synthesised narratively, with greater weight given to controlled studies. MAIN RESULTS: We identified 30 studies of which 13 were controlled. Giving retailers information was less effective in reducing illegal sales than active enforcement and/or multicomponent educational strategies. No strategy achieved complete, sustained compliance. In three controlled trials, there was little effect of intervention on youth perceptions of access or prevalence of smoking. REVIEWER'S CONCLUSIONS: Interventions with retailers can lead to large decreases in the number of outlets selling tobacco to youths. However, few of the communities studied in this review achieved sustained levels of high compliance. This may explain why there is limited evidence for an effect of intervention on youth perception of ease of access to tobacco, and on smoking behaviour. PMID- 11869603 TI - Laparoscopic versus open surgery for suspected appendicitis. AB - BACKGROUND: Laparoscopic surgery has been proposed to have diagnostic and therapeutic advantages over conventional surgery. OBJECTIVES: To compare the diagnostic and therapeutic effects of laparoscopic and conventional 'open' surgery in the treatment of suspected acute appendicitis. SEARCH STRATEGY: We searched for original articles and abstracts published until end of 2000. As main search tools we employed the Cochrane Controlled Trials Register (CCTR), MEDLINE, EMBASE and SciSearch. CCTR and MEDLINE searches were repeated until 10 October 2001, all other databases were searched 10 October 2000. We also handsearched the congress proceedings of endoscopic surgical societies. SELECTION CRITERIA: We included clinical trials that assessed either: (1) Therapeutic effects of laparoscopic appendectomy (LA) versus open appendectomy (OA) in adults, (2) Therapeutic effects of LA versus OA in children, (3) Diagnostic effects of diagnostic laparoscopy (LAP) followed by LA or OA if necessary versus immediate OA, (4) Therapeutic effects of diagnostic laparoscopy (LAP) followed by OA if necessary versus immediate OA. We included only randomized studies and excluded those with unconcealed allocation. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed each study's eligibility and quality. One reviewer extracted the data, 10% of which were later cross-checked by a second reviewer. Abstract authors and authors of articles lacking important information on trial design or results were contacted. MAIN RESULTS: We included 45 studies, of which 39 compared LA (with or without diagnostic laparoscopy) vs. OA in adults. Wound infections were about half as likely (Peto OR 0.47; 95%-CI 0.36 to 0.62) after LA than after OA, but intraabdominal abscesses were increased nearly threefold after LA (Peto OR 2.77; 95%-CI 1.61 to 4.77). The duration of surgery was 14 minutes (95%-CI 10 to 19) longer for LA. Pain on day 1 after surgery was reduced after LA by 8 mm (95%-CI 3 to 13 mm) on a 100 mm VAS. Hospital stay was reduced by 0.7 days (95%-CI 0.4 to 1.0). Return to normal activity, work, and sport were 6 days (95%-CI 4 to 8), 3 days (1 to 5), and 7 days (3 to 12) earlier after LA than after OA. While the operation costs of LA were significantly higher than that of OA, the costs outside hospital were reduced. Strong heterogeneity was found for most outcomes, but not for wound infections and intraabdominal abscesses. In children, much less data were available, but the result do not seem to be much different when compared to adults. Pain which was measured blindly in two paediatric trials, was similar after LA and OA (-1 mm VAS; 95%-CI -8 to +7 mm). In trials on unselected patients, diagnostic laparoscopy led to large but variable reductions in the rate of negative appendectomies (RR 0.21; 95%-CI 0.13 to 0.33). In parallel, the rate of unestablished diagnoses was significantly decreased after laparoscopy (RR 0.34; 95%-CI 0.22 to 0.53). In fertile women, these effects were even more pronounced: rate of negative appendectomies: RR 0.19; 95%-CI 0.11 to 0.34; rate of patients without a final diagnosis established: RR 0.24; 95%-CI 0.15 to 0.38. REVIEWER'S CONCLUSIONS: In those clinical settings where surgical expertise and equipment are available and affordable, diagnostic laparoscopy and LA (either in combination or separately) seem to have various advantages over OA. Some of the clinical effects of LA, however, are small and of limited clinical relevance. In spite of the mediocre quality of the available research data, we would generally recommend to use laparoscopy and LA in patients with suspected appendicitis unless laparoscopy itself is contraindicated or not feasible. In gangrenous or perforated cases, however, LA may possibly carry a higher risk of intraabdominal infections. PMID- 11869604 TI - Decongestants and antihistamines for acute otitis media in children. AB - BACKGROUND: Acute otitis media (AOM) is a common and important source of morbidity in children, although most cases resolve spontaneously. While frequently recommended, decongestant and antihistamine therapy is of unclear benefit. OBJECTIVES: To determine the efficacy of decongestant and antihistamine therapy in children with AOM on outcomes of AOM resolution, medication side effects, and complications of AOM. SEARCH STRATEGY: Comprehensive search of Cochrane's Controlled Trials Registry, Medline and Embase was conducted. Bibliographic review and requests for information from study authors and pharmaceutical companies supplemented this. SELECTION CRITERIA: Randomized controlled trials evaluating decongestant (DC) or antihistamine (AH) treatment for children with AOM were included. Patient-oriented outcomes were considered most relevant. There were no quality or language restrictions. DATA COLLECTION AND ANALYSIS: Investigators independently evaluated studies for inclusion, performed validity assessments, and completed data extraction. Dichotomous data were pooled to generate relative risks and numbers needed to treat, and homogeneity was assessed using approximate chi-square tests. MAIN RESULTS: For the combined control groups, healing rates at 2 weeks were high, with rates of persistent AOM <23%. No additional benefit was demonstrated from intervention subgroupings DC, AH, or any medication (DC and/or AH). Only the combined treatment (DC + AH) group demonstrated statistically lower rates of persistent AOM at the 2-week period (RR 0.76, 95% CI, NNT 10.5, 95% CI). No benefit was found for other outcomes including early or late cure rates, symptom resolution, prevention of surgery or other complications. There was an increased risk of medication side effects for those receiving an intervention, which reached statistical significance for the "any medication" and decongestant groupings (NNH 16.6, 14.3 respectively, 95% CI ). Validity subanalyses demonstrated that lower quality studies found benefit, but analysis of those studies with higher validity scores found no benefit to treatment. REVIEWER'S CONCLUSIONS: Given lack of benefit and increased risk of side effects, these data do not support the use of decongestant, antihistamine, or combined DC/AH treatment in children with AOM. The small statistical benefit found in the combination medication group is of small clinical significance and study design may be biasing the results. PMID- 11869605 TI - Injection sclerotherapy for varicose veins. AB - BACKGROUND: Injection sclerotherapy for varicose veins has been used widely since 1963, following popularisation of the technique by Fegan. The treatment aims to obliterate the lumen of varicose veins or thread veins, however, there is limited evidence regarding its efficacy. OBJECTIVES: To determine whether sclerotherapy is effective in terms of symptomatic improvement and cosmetic appearance; has an acceptable complication rate; and to define rates of symptomatic or cosmetic varicose vein recurrence following sclerotherapy. SEARCH STRATEGY: Publications describing randomised controlled trials (RCTs) of injection sclerotherapy for varicose veins (excluding comparisons with surgery) were sought through EMBASE and MEDLINE (from inception to March 2001) and hand-searching relevant journals, using the search strategy described by the Cochrane Peripheral Vascular Diseases Review Group. Bibliographies of papers identified were examined for further RCTs. Manufacturers of sclerosants were contacted for further trial information. SELECTION CRITERIA: RCTs of injection sclerotherapy versus graduated compression stockings or 'observation', or comparing different sclerosants, doses and post compression bandaging techniques on patients with symptomatic and/or cosmetic varicose veins or thread veins were considered for inclusion in the review. DATA COLLECTION AND ANALYSIS: Ten studies were included in the analysis. These compared: sodium tetradecyl sulphate (STD) versus an alternative sclerosant; sclerosant with or without local anaesthetic; application of Molefoam versus Sorbo pads to injection sites; elastic compression bandaging versus conventional bandaging; and short-term bandaging versus standard bandaging. Data were abstracted by both authors. MAIN RESULTS: No RCTs compared sclerotherapy to graduated compression stockings or other non-surgical treatments. Two studies compared STD to alternative sclerosants and found no significant differences in outcome or complication rates. Adding local anaesthetic to sclerosant reduced the pain from injection (one study) but had no other effects. Comparison of Molefoam and Sorbo pad pressure dressings found no difference in outcome for erythema (redness) or successful sclerosis. The degree and duration of elastic compression had no significant effect on varicose vein recurrence rates, cosmetic appearance or symptomatic improvement. Increased compression prevented slipping of dressings but caused increased discomfort, as did increasing duration of compression. REVIEWER'S CONCLUSIONS: Evidence from RCTs suggests that type of sclerosant, local pressure dressing, degree and length of compression have no significant effect on the efficacy of sclerotherapy for varicose veins. This supports the current place of sclerotherapy in modern clinical practice, which is usually limited to treatment of recurrent varicose veins following surgery, and thread veins. A comparison of surgery versus sclerotherapy would be valuable. PMID- 11869606 TI - Pentoxifylline for treating venous leg ulcers. AB - BACKGROUND: Healing of venous leg ulcers is improved by the use of compression bandaging but some venous ulcers remain unhealed, and some people are unsuitable for compression therapy. Pentoxifylline, a drug which helps blood flow, has been used to treat venous leg ulcers but to date there has been no systematic review. OBJECTIVES: To assess the effects of pentoxifylline (oxpentifylline or Trental 400) for treating venous leg ulcers, compared with placebo, or other therapies, in the presence or absence of compression therapy. SEARCH STRATEGY: We searched the Cochrane Peripheral Vascular Diseases and Wounds Groups specialised registers (date of last search was May 2001), and reference lists of relevant articles. We hand searched relevant journals and conference proceedings, and contacted Hoechst (the manufacturer of the drug) and experts in the field. SELECTION CRITERIA: Randomised trials comparing pentoxifylline with placebo or other therapy in the presence or absence of compression, in patients with venous leg ulcers. DATA COLLECTION AND ANALYSIS: Details from eligible trials were extracted and summarised by one reviewer using a coding sheet. Data extraction was independently verified by one other reviewer. MAIN RESULTS: Nine trials involving 572 adults were included. The quality of trials was variable. Eight trials compared pentoxifylline with placebo; in five of these trials patients received compression therapy. In one trial pentoxifylline was compared with defibrotide in patients who also received compression. Combining eight trials that compared pentoxifylline with placebo (with or without compression) demonstrated that pentoxifylline is more effective than placebo in terms of complete ulcer healing or significant improvement (relative risk of healing with pentoxifylline compared with placebo 1.41, 95% confidence interval 1.19 -1.66). Pentoxifylline plus compression is more effective than placebo plus compression (relative risk of healing with pentoxifylline 1.30, 95% confidence interval 1.10-1.54). A comparison between pentoxifylline and defibrotide found no difference in healing rates. More adverse effects were reported in patients receiving pentoxifylline, although this was not statistically significant (relative risk of adverse effects with pentoxifylline 1.25, 95% confidence interval 0.87-1.80). Nearly half of the reported adverse effects were gastrointestinal. REVIEWER'S CONCLUSIONS: Pentoxifylline appears to be an effective adjunct to compression bandaging for treating venous ulcers. There was no cost effectiveness data available and healthcare commissioners may therefore conclude that it not be considered a routine adjunct. Pentoxifylline in the absence of compression may be effective for treating venous leg ulcers, although the evidence should be cautiously interpreted. The majority of adverse effects are gastrointestinal disturbances (indigestion, diarrhoea and nausea). PMID- 11869607 TI - Pre-conception and antenatal screening for the fragile site on the X-chromosome. AB - BACKGROUND: Fragile X is the most common cause of mental retardation after Down syndrome. It is the commonest inherited cause of mental retardation, and results from a dynamic mutation in a gene on the long arm of the X chromosome. Various strategies are used for prenatal screening. OBJECTIVES: To determine whether pre conceptual or antenatal screening for Fragile X carrier status in apparently low risk women confers any additional benefit over the existing practice of offering testing to women thought to be at increased risk. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register (November 2001), the Cochrane Controlled Trials Register (The Cochrane Library Issue 3, 2001), MEDLINE (1980 to 2001), and reference lists of articles. SELECTION CRITERIA: Randomised clinical trials comparing women being tested regardless of family history (intervention group) with women tested only when there is a family history of either fragile X and/or other undiagnosed mental illness/impairment (control group). DATA COLLECTION AND ANALYSIS: Three reviewers independently assessed trial quality and extracted data. MAIN RESULTS: No trials were included. REVIEWER'S CONCLUSIONS: No information is available from randomised trials to indicate whether routine pre-conceptual or antenatal screening for fragile X carrier status confers any benefit over testing women thought to be at increased risk. PMID- 11869608 TI - Diuretics acting on the distal renal tubule for preterm infants with (or developing) chronic lung disease. AB - OBJECTIVES: The aim of this review is to assess the risks and benefits of diuretics acting on distal segments of the renal tubule (distal diuretics) in preterm infants with or developing chronic lung disease (CLD). Primary objectives are to assess changes in need for oxygen or ventilatory support and effects on long-term outcome, and secondary objectives are to assess changes in pulmonary mechanics and potential complications of therapy. SEARCH STRATEGY: We used the standard method of the Cochrane Neonatal Review Group. We searched MEDLINE (1966 November 2001), EMBASE (1974-November 2001) and the Cochrane Controlled Trials Register (CCTR) (Cochrane Library Issue 4, 2001). In addition, we hand searched several abstract books of national and international American and European Societies. SELECTION CRITERIA: We included in this analysis trials in which preterm infants with or developing CLD and at least five days of age were all randomly allocated to receive a distal diuretic (i.e., a diuretic acting on the distal renal tubule). Eligible studies needed to assess at least one of the outcome variables defined a priori for this systematic review. Primary outcome variables included changes in need for respiratory support and oxygen supplementation, mortality, bronchopulmonary dysplasia (BPD), death or BPD, chronic lung disease at 36 weeks of postconceptional age (gestational age + postnatal age), length of stay, and number of rehospitalizations during the first year of life. Secondary outcome variables included pulmonary mechanics and potential complications of therapy. DATA COLLECTION AND ANALYSIS: We used the standard method for the Cochrane Collaboration which is described in the Cochrane Collaboration Handbook. Two investigators extracted, assessed and coded separately all data for each study, using a form that was designed specifically for this review. Any disagreement was resolved by discussion. We combined parallel and cross-over trials and, whenever possible, transformed baseline and final outcome data measured on a continuous scale into change scores using Follmann's formula. MAIN RESULTS: Of six studies fulfilling entry criteria, most focused on pathophysiological parameters and did not assess effects on important clinical outcomes defined in this review, or the potential complications of diuretic therapy. In preterm infants > 3 weeks of age with CLD, a four-week treatment with thiazide and spironolactone improved lung compliance and reduced the need for furosemide. Thiazide and spironolactone decreased the risk of death and tended to decrease the risk for lack of extubation after 8 weeks in intubated infants who did not have access to corticosteroids, bronchodilators or aminophylline. However, there is little or no evidence to support any benefit of diuretic administration on need for ventilatory support, length of hospital stay, or long-term outcome in patients receiving current therapy. There is no evidence to support the hypothesis that adding spironolactone to thiazide or that adding metolazone to furosemide improves the outcome of preterm infants with CLD. REVIEWER'S CONCLUSIONS: In preterm infants > 3 weeks of age with CLD, acute and chronic administration of distal diuretics improve pulmonary mechanics. Studies are needed to assess (1) whether thiazide administration improves mortality, duration of oxygen dependency, ventilator dependency, length of hospital stay and long-term outcome in patients exposed to corticosteroids and bronchodilators (2) whether adding spironolactone to thiazides or adding metolazone to furosemide has any beneficial effect. PMID- 11869609 TI - Orthotic devices for the treatment of tennis elbow. AB - BACKGROUND: Lateral epicondylitis (tennis elbow) is a frequently reported condition. A wide variety of treatment strategies has been described. As of yet, no optimal strategy has been identified. OBJECTIVES: To assess the effectiveness of orthotic devices for the treatment of tennis elbow. SEARCH STRATEGY: We searched Medline, Embase, CINAHL, the Cochrane Controlled Trial Register, Current Contents up to May 1999 and reference lists from all retrieved articles. Experts on the subjects were approached for additional trials. SELECTION CRITERIA: All randomised clinical trials (RCT) describing individuals with diagnosed lateral epicondylitis and comparing the use of an orthotic device as a treatment strategy were evaluated for inclusion. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed the validity of the included trials and extracted data on relevant outcome measures. Dichotomous outcomes were expressed as Relative Risks (RRs) and continuous outcomes as Standardised Mean Differences (SMD), both with corresponding 95% confidence intervals (95% CI). Statistical pooling and subgroup analyses were intended MAIN RESULTS: Five RCTs (N per group 7-49) were included. Validity score ranged from 3-9 positive items out of 11. Subgroup analyses were not performed due to the small number of trials. The limited number of included trials present few outcome measures and limited long-term results. Pooling was not possible due to large heterogeneity amongst trials. REVIEWER'S CONCLUSIONS: No definitive conclusions can be drawn concerning effectiveness of orthotic devices for lateral epicondylitis. More well-designed and well-conducted RCTs of sufficient power are warranted. PMID- 11869610 TI - Antibiotics for trachoma. AB - BACKGROUND: Trachoma is the world's leading cause of preventable blindness. In 1997 the World Health Organization launched an initiative on trachoma control based on the 'SAFE' strategy (surgery, antibiotics, facial cleanliness and environmental improvement). OBJECTIVES: The aim of this review is to assess the evidence supporting the antibiotic arm of the SAFE strategy by assessing the effects of antibiotics on both active trachoma (primary objective) and on Chlamydia trachomatis infection of the conjunctiva (secondary objective). SEARCH STRATEGY: We searched The Cochrane Controlled Trials Register - CENTRAL/CCTR, which contains the Cochrane Eyes and Vision Group specialised register (Cochrane Library Issue 3, 2001), MEDLINE (1966 to August 2001), and EMBASE (1980 to September 2001). We used the Science Citation Index to look for articles that cited the included studies. We searched the reference lists of identified articles and we contacted authors and experts for details of further relevant studies. SELECTION CRITERIA: We included only randomised trials that satisfied either of two criteria: (a) trials in which topical or oral administration of an antibiotic was compared to placebo or no treatment in people with trachoma, (b) trials in which a topical antibiotic was compared with an oral antibiotic in people with trachoma. A subdivision of particular interest was of trials in which topical tetracycline/chlortetracycline was compared with oral azithromycin, as these are the two World Health Organization recommended treatments. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed trial quality and extracted data. We contacted investigators for missing data. MAIN RESULTS: We found 15 studies that randomised a total of 8678 participants. For both outcomes (active trachoma and laboratory evidence of infection) the results of the chi square tests suggested that there was significant statistical heterogeneity among the trials. There was also marked clinical heterogeneity. No summary statistics were calculated and we therefore present a narrative summary of the results. For the comparisons of oral or topical antibiotic against placebo/no treatment, the data are consistent with there being no effect of antibiotics but are suggestive of a lowering of the point prevalence of relative risk of both active disease and laboratory evidence of infection at three and 12 months after treatment. For the comparison of oral against topical antibiotics the results suggest that oral treatment is neither more nor less effective than topical treatment. REVIEWER'S CONCLUSIONS: There is some evidence that antibiotics reduce active trachoma but results are not consistent and cannot be pooled. PMID- 11869611 TI - Physical methods for preventing deep vein thrombosis in stroke. AB - BACKGROUND: Deep vein thrombosis (DVT) and resulting pulmonary embolism (PE) are uncommon but important complications of stroke. There is good evidence that anticoagulants can reduce the risk of DVT and PE after stroke, but this benefit is offset by a small but definite risk of serious haemorrhages. Physical methods to prevent DVT and PE (such as compression stockings applied to the legs) are not associated with any bleeding risk and are effective in some categories of medical and surgical patients. We sought to assess their effects in stroke patients. OBJECTIVES: To assess the effectiveness and safety of physical methods of preventing the onset of deep vein thrombosis and fatal or non fatal pulmonary embolism in patients with recent stroke. SEARCH STRATEGY: We searched the Cochrane Stroke Group Trials Register (last searched October 2001). In addition we searched the following electronic bibliographic databases: Cochrane Controlled Trials Register (1999, Issue 3), MEDLINE (1966- Jan 2001), EMBASE (1980- Jan 2001) and CINAHL (1982-May 1999). The reference lists of all relevant papers were screened for additional trials. SELECTION CRITERIA: All completed randomised unconfounded trials or controlled clinical trials comparing physical methods in patients allocated to receive physical methods, applied within one week of onset of stroke, with patients allocated to no physical methods. DATA COLLECTION AND ANALYSIS: Two reviewers independently searched for relevant trials and three others independently checked the results. MAIN RESULTS: We identified two small trials which included 123 patients. In one trial of 97 patients, compression stockings were associated with a non significant trend towards a reduction in DVT detected by Doppler ultrasound. In one trial of 26 patients, an intermittent pneumatic compression device was not associated with a significant reduction in DVT detected by 125-I-fibrinogen scanning. Overall, physical methods were not associated with a significant reduction in DVT (Odds ratio 0.59, 95%.CI 0.24 1.48) or death (Odds ratio 5.06, 95% CI 0.96-26.78). REVIEWER'S CONCLUSIONS: There is insufficient evidence from randomised trials to support the routine use of physical methods for preventing DVT in acute stroke. PMID- 11869612 TI - Lubeluzole for acute ischaemic stroke. AB - BACKGROUND: Experimental studies have shown that ischaemic insults cause excess release of excitatory amino acid (EAA) neurotransmitters, particularly glutamate. Glutamate re-uptake is impaired under ischaemic conditions. In preclinical models of stroke, antagonists of excitatory amino acids or of glutamate release protect against ischaemic injury, even when administered after the ischaemic insult. Lubeluzole is a benzothiazole derivative that has shown neuroprotective properties in different experimental models inhibiting glutamate release, nitric oxide (NO) synthesis and blocking voltage-gated Na+ and Ca2+ ion channels. OBJECTIVES: The objective of this review is to assess the effectiveness and safety of lubeluzole given in the acute phase of acute ischaemic stroke. SEARCH STRATEGY: The Cochrane Stroke Group trials register was searched. Additional searches of Cochrane Controlled Trials Register (CENTRAL/CCTR), Medline, Embase, Pascal BioMed (1996-2001) and Current Contents CCSearch reg 7 Editions (1996 2001) were made to supplement the Stroke Group general strategy. We contacted Janssen Research Foundation to identify further studies. SELECTION CRITERIA: All randomised unconfounded trials comparing intravenous lubeluzole with placebo or open control in patients with a clinical syndrome definitely considered as an acute stroke in whom CT scanning showed an infarct or was normal. DATA COLLECTION AND ANALYSIS: Two reviewers independently selected trials for inclusion, assessed trial quality and extracted the data. MAIN RESULTS: Five trials involving a total of 3,510 patients were included. The quality of the trials did not vary considerably. Sensitivity/subgroups analysis was not completely performed because of lack of data. Lubeluzole given at the doses of 5, 10 and 20 mg/day for 5 days was tested against a placebo-control group. There was no evidence that Lubeluzole given at any dose either reduced the odds of death from all causes (OR=0.93, 95% CI 0.79-1.09) or reduced the odds of being dead or dependent at the end of follow up (OR=1.04, 95% CI 0.91-1.19). On the other hand, given at any dose, Lubeluzole was associated with a significant excess of heart-conduction disorders (Q-T prolonged > 450 msec) at the end of follow-up (OR=1.43, 95% CI 1.09-1.87). REVIEWER'S CONCLUSIONS: Lubeluzole, given in the acute phase of ischaemic stroke, is not associated with a significant reduction of death or dependency at the end of scheduled follow-up period but seems to be associated with a significant increase of heart-conduction disorders (Q-T prolonged >450 msec). PMID- 11869613 TI - Corticosteroids for Bell's palsy (idiopathic facial paralysis). AB - BACKGROUND: Inflammation and oedema of the facial nerve are implicated in causing Bell's palsy. Corticosteroids have a potent anti-inflammatory action which should minimise nerve damage and thereby improve the outcome of patients suffering from this condition. OBJECTIVES: The objective of this review was to assess the effect of steroid therapy in the recovery of patients with Bell's palsy. SEARCH STRATEGY: We searched the Cochrane Neuromuscular Disease Group register for randomised trials, as well as MEDLINE, EMBASE and LILACS (to December 2000). We contacted known experts in the field to identify additional published or unpublished trials. SELECTION CRITERIA: Randomised trials comparing different routes of administration and dosage schemes of corticosteroid or adrenocorticotrophic hormone therapy versus a control group where no therapy considered effective for this condition was administered, unless it was also given in a similar way to the experimental group. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed eligibility, trial quality, and extracted the data. MAIN RESULTS: Three trials with a total of 117 patients were included. One trial compared cortisone acetate with placebo; one compared prednisone plus vitamins, with vitamins alone; and one, not-placebo controlled, tested the efficacy of methylprednisolone. Allocation concealment was appropriate in two trials, and the data reported allowed an intention-to-treat analysis. Overall 13/59 (22%) of the patients allocated to steroid therapy had incomplete recovery of facial motor function six months after randomisation, compared with 15/58 (26%) in the control group. This reduction was not significant (relative risk 0.86, 95% confidence interval 0.47 to 1.59). The reduction in the proportion of patients with cosmetically disabling sequelae six months after randomisation was also not significant (relative risk 0.86, 95% confidence interval 0.38 to 1.98). REVIEWER'S CONCLUSIONS: The available evidence from randomised controlled trials does not show significant benefit from treating Bell's palsy with corticosteroids. More randomised controlled trials with a greater number of patients are needed to determine reliably whether there is real benefit (or harm) from the use of steroid therapy in patients with Bell's palsy. PMID- 11869614 TI - Benzodiazepines for neuroleptic-induced acute akathisia. AB - BACKGROUND: Neuroleptic-induced akathisia is one of the most common and distressing early-onset adverse effects of antipsychotic drugs, being associated with poor compliance with treatment, and thus, ultimately, to an increase risk of relapse. This review assesses the role of benzodiazepines in the pharmacological treatment of this problem. OBJECTIVES: To determine the effects of benzodiazepines versus placebo for people with neuroleptic-induced acute akathisia. SEARCH STRATEGY: Biological Abstracts (January 1982-March 1999), The Cochrane Library (Issue 3 1999), The Cochrane Schizophrenia Group's Register (May 2001), EMBASE (January 1980-March 1999), LILACS (January 1982-March 1999), MEDLINE (January 1964-March 1999), PsycLIT (January 1974-March 1999), and SCISEARCH were searched. Further references were sought from published trials and their authors. SELECTION CRITERIA: All randomised clinical trials comparing benzodiazepines with placebo for people with antipsychotic-induced acute akathisia. DATA COLLECTION AND ANALYSIS: Two reviewers, working independently, selected, quality assessed and extracted data. These data were then analysed on an intention-to-treat basis. For homogeneous dichotomous data the fixed effects relative risk (RR), the 95% confidence intervals (CI) and, where appropriate, the number needed to treat (NNT) were calculated on an intention-to-treat basis. For continuous data, reviewers calculated weighted mean differences. MAIN RESULTS: Two small (total N=27) randomised controlled trials were included. By seven to 14 days, there was a reduction in symptoms for those patients receiving clonazepam compared with placebo (2 RCTs, N=26, RR 0.09 CI 0.01 to 0.6, NNT 1.2 CI 0.9 to 1.5). No significant difference was found for adverse events (2 RCTs, N=26, RR 3.00 CI 0.2 to 62) or the need for anticholinergic medication (2 RCTs, N=26, RR 1.56 CI 0.9 to 2.7). No one left the two studies early. Data on mental, social and family outcomes could not be pooled and there was little or no data on user satisfaction, deaths, violence, criminal behaviour and costs. REVIEWER'S CONCLUSIONS: Over a short follow-up period, the use of benzodiazepines may reduce the symptoms of antipsychotic-induced acute akathisia. This review highlights the need for well designed, conducted and reported clinical trials to address the claims of open studies. PMID- 11869615 TI - Interventions for treating oral candidiasis for patients with cancer receiving treatment. AB - BACKGROUND: Treatment of cancer is increasingly effective but is associated with short and long-term side effects. Oral side effects, including oral candidiasis, remain a major source of illness despite the use of a variety of agents to treat them. OBJECTIVES: To assess the effectiveness of interventions for the treatment of oral candidiasis for patients with cancer receiving chemotherapy and or radiotherapy. SEARCH STRATEGY: Computerised searches of Cochrane Oral Health Group Specialised Register, CCTR, MEDLINE and EMBASE were undertaken. Reference lists from relevant articles were searched and the authors of eligible trials were contacted to identify trials and obtain additional information. Date of the most recent searches May 2001: (CCTR 2001, issue 3) SELECTION CRITERIA: All randomised controlled trials comparing agents prescribed to treat oral candidiasis in people receiving chemotherapy or radiotherapy for cancer. The outcomes were eradication of oral candidiasis, dysphagia, systemic infection, amount of analgesia, length of hospitalisation, cost and patient quality of life. DATA COLLECTION AND ANALYSIS: Data were independently extracted, in duplicate, by two reviewers. Authors were contacted for details of randomisation and withdrawals and a quality assessment was carried out. The Cochrane Oral Health Group statistical guidelines were followed and relative risk values calculated using random effects models where significant heterogeneity was detected (P<0.1). MAIN RESULTS: Eight trials involving 418 patients, satisfied the inclusion criteria and are included in this review. Only two agents, each in single trials, were found to be effective for eradicating oral candidiasis. A drug absorbed from the gastrointestinal tract, ketoconazole, was more beneficial than placebo in eradicating oral candidiasis (RR=0.35 95%CI 0.20 to 0.61) and clotrimazole, at a higher dose of 50mg was more effective than a lower 10mg dose in eradicating oral candidiasis, when assessed mycologically (RR=0.47 95%CI 0.25 to 0.89). Another trial demonstrated no difference between a 10mg dose of the partially absorbed drug, clotrimazole, and placebo. No differences were found when comparing different absorbed drugs; and comparing absorbed drugs with drugs which are not absorbed. REVIEWER'S CONCLUSIONS: There is weak and unreliable evidence that the absorbed drug, ketoconazole, may eradicate oral candidiasis and that a higher dose of the partially absorbed drug, clotrimazole, may give greater benefit than a lower 10mg dose, however, researchers may wish to prevent rather than treat oral candidiasis. Further well designed, placebo-controlled trials assessing the effectiveness of old and new interventions for treating oral candidiasis are needed. PMID- 11869616 TI - Interventions for treating oral mucositis for patients with cancer receiving treatment. AB - BACKGROUND: Treatment of cancer is increasingly effective but associated with short and long-term side effects. Oral side effects, including oral mucositis (ulceration), remain a major source of illness despite the use of a variety of agents to treat them. OBJECTIVES: To assess the effectiveness of interventions for treating oral mucositis or its associated pain in patients with cancer receiving chemotherapy and/or radiotherapy. SEARCH STRATEGY: Computerised searches of Cochrane Oral Health Group Specialised Register, CCTR, MEDLINE and EMBASE were undertaken. Reference lists from relevant articles were searched. Authors of eligible trials were contacted to identify trials and obtain additional information. Date of most recent searches: May 2001 (CCTR 2001, issue 3) SELECTION CRITERIA: All randomised controlled trials comparing agents prescribed to treat oral mucositis in people receiving chemotherapy and/or radiotherapy. Outcomes were oral mucositis, oral pain, dysphagia, systemic infection, amount of analgesia, length of hospitalisation, cost and quality of life. DATA COLLECTION AND ANALYSIS: Data were independently extracted, in duplicate, by two reviewers. Authors were contacted for details of randomisation, blindness and withdrawals. Quality assessment was carried out on these three criteria. Cochrane Oral Health Group statistical guidelines were followed and relative risk values calculated using fixed effects models as no significant heterogeneity was detected (P>0.1). MAIN RESULTS: Fifteen trials involving 876 patients satisfied the inclusion criteria. Two agents, each in single trials, were found to be effective for improving (allopurinol RR=0.63 95%CI 0.42 to 0.96) or eradicating mucositis (allopurinol RR=0.59 95%CI 0.42 to 0.84; vitamin E RR=0.38 95%CI 0.14 to 0.97). The following agents were not found to be effective: benzydamine HCl, sucralfate, tetrachlorodecaoxide, chlorhexidine and "magic" (lidocaine solution, diphenhydramine hydrochloride and aluminum hydroxide suspension). Three trials compared patient controlled analgesia (PCA) to the continuous infusion method for controlling pain. There was no evidence of a difference, however, less opiate was used per hour for PCA. One trial demonstrated that pharmacokinetically based analgesia (PKPCA) reduced pain compared with PCA, however more opiate was used with PKCA. REVIEWER'S CONCLUSIONS: There is weak and unreliable evidence that allopurinol mouthwash and vitamin E improves or eradicates mucositis. There is no evidence that patient controlled analgesia (PCA) is better than continuous infusion method for controlling pain, however, less opiate was used per hour for PCA. Further, well designed, placebo-controlled trials assessing the effectiveness of allopurinol mouthwash, vitamin E and new interventions for treating mucositis are needed. PMID- 11869617 TI - Interventions for acute non-arteritic central retinal artery occlusion. AB - BACKGROUND: Acute central retinal artery occlusion occurs as a sudden interruption of the blood supply to the retina and results in an almost complete loss of vision in the affected eye. There is no generally agreed treatment regimen although a number of therapeutic interventions have been proposed. OBJECTIVES: The objective of this review is to examine the effects of treatments used for acute non-arteritic central retinal artery occlusion. SEARCH STRATEGY: We searched the Cochrane Controlled Trials Register - CENTRAL/CCTR, which contains the Cochrane Eyes and Vision Group specialised register (Cochrane Library Issue 4, 2001), MEDLINE (1966 to August 2001) and EMBASE (1980 to September 2001). We searched the reference lists of relevant papers for trials. SELECTION CRITERIA: We included only randomised controlled trials in which one treatment aimed to re-establish blood supply to the retina in people with acute central retinal artery occlusion was compared to another treatment. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed the search results for trials to be included in the review. Discrepancies were resolved by discussion. MAIN RESULTS: We found no randomised controlled trials that met our inclusion criteria. REVIEWER'S CONCLUSIONS: There is currently not enough evidence to decide which, if any, interventions for acute non-arteritic central retinal artery occlusion would result in any beneficial or harmful effect. Well designed randomised controlled trials are needed to establish the most effective treatment. PMID- 11869618 TI - Vitamin K antagonists or low-molecular-weight heparin for the long term treatment of symptomatic venous thromboembolism. AB - BACKGROUND: People with venous thromboembolism are generally treated for five days with intravenous unfractionated heparin or subcutaneous low-molecular-weight heparin followed by three months of vitamin K antagonists treatment. Treatment with vitamin K antagonists requires regular laboratory measurements and some patients have contraindications for treatment. OBJECTIVES: To evaluate the efficacy and safety of long-term treatment of venous thromboembolism with low molecular-weight heparins compared to vitamin K antagonists. SEARCH STRATEGY: Searches of MEDLINE, EMBASE and ISI Web of Science, the Specialised Trials Register of the Cochrane Peripheral Vascular Disease Group and the Cochrane Controlled Trials Register were made and relevant journals were hand-searched. Additional trials were sought through communication with colleagues and pharmaceutical companies. SELECTION CRITERIA: Two reviewers evaluated studies independently for methodological quality. DATA COLLECTION AND ANALYSIS: Two reviewers extracted data independently. Primary analysis concerned all trial participants during the period of randomized treatment. Separate analyses were performed for category I and category II studies; i.e. studies using similar treatments initially in both study arms, and those that did not; and the different periods of follow-up. MAIN RESULTS: All seven studies fulfilling our criteria combined, a statistically non-significant reduction in the risk of recurrent venous thromboembolism favoring low-molecular-weight heparin treatment (OR 0.70; 95% CI [0.42, 1.16]) was found. Analysis of pooled data for category I studies showed a non-significant reduction in the risk of recurrent venous thromboembolism favoring low-molecular-weight heparin treatment (OR 0.75; 95% CI [0.40, 1.39]). Omitting a potentially-confounded study, a statistically non significant reduction in the risk of recurrent venous thromboembolism favoring vitamin K antagonist treatment remained (OR 1.95; 95% CI [0.74, 5.19]). All studies combined, the difference in bleeding significantly favored treatment with low-molecular-weight heparin (OR 0.38; 95% CI [0.15, 0.94]), however, considering only category I studies a non-significant trend favoring low-molecular-weight heparin remained (OR 0.80; 95% CI [0.21, 3.00]). No difference was observed in mortality (OR 1.13; 95% CI [0.47, 2.69]). REVIEWER'S CONCLUSIONS: Low-molecular weight heparins are possibly as effective as vitamin K antagonists in preventing symptomatic venous thromboembolism after an episode of symptomatic deep venous thrombosis, but are much more expensive. Treatment with low-molecular-weight heparin is significantly safer than treatment with vitamin K antagonists and is possibly a safe alternative in some patients; especially those in geographically inaccessible places, reluctant to visit the thrombosis service regularly, or with contraindications to vitamin K antagonists. However, treatment with vitamin K antagonists remains the treatment of choice for the majority of patients. PMID- 11869619 TI - Hypertonic versus isotonic crystalloid for fluid resuscitation in critically ill patients. AB - BACKGROUND: Hypertonic solutions are considered to have a greater ability to expand blood volume and thus elevate blood pressure and can be administered as a small volume infusion over a short time period. On the other hand, the use of hypertonic solutions for volume replacement may also have important disadvantages. OBJECTIVES: To determine whether hypertonic crystalloid decreases mortality in patients with hypovolaemia with and without head injuries. SEARCH STRATEGY: We searched MEDLINE, EMBASE, The Cochrane Controlled Trials Register and the Specialised register of the Injuries Group. We checked reference lists of all articles identified and searched the National Research Register. SELECTION CRITERIA: Randomised trials comparing hypertonic to isotonic crystalloid in patients with trauma, burns or undergoing surgery. DATA COLLECTION AND ANALYSIS: Two authors independently extracted the data and assessed the quality of the trials. MAIN RESULTS: Seventeen trials were identified with a total of 869 participants. Data on death were obtained in twelve of the studies. Only one trial reported data on disability. The pooled RR for death in trauma patients was 0.84 (95% CI 0.61-1.16), in patients with burns 1.49 (95% CI 0.56-3.95), and in patients undergoing surgery 0.62 (95% cI 0.08-4.57). In the one trial that gave data on disability using the Glasgow Outcome Scale the relative risk was 0.99 (95% CI 0.06-15.93). REVIEWER'S CONCLUSIONS: This review does not give us enough data to be able to say whether hypertonic crystalloid is better than isotonic crystalloid for the resuscitation of patients with trauma, burns, or those undergoing surgery. However, the confidence intervals are wide and do not exclude clinically significant differences. Further trials are needed comparing hypertonic to isotonic crystalloid. Trials need to be large enough to detect a clinically important difference. PMID- 11869620 TI - Corticosteroids for chronic inflammatory demyelinating polyradiculoneuropathy. AB - BACKGROUND: Chronic inflammatory demyelinating polyradiculoneuropathy is a peripheral neuropathy caused by peripheral nerve inflammation probably due to autoimmunity and would be expected to benefit from corticosteroids. Non randomised studies suggest that corticosteroids are often beneficial. OBJECTIVES: To evaluate the efficacy of corticosteroids for treating chronic inflammatory demyelinating polyradiculoneuropathy. SEARCH STRATEGY: Search of the Cochrane Neuromuscular Disease Group register for randomised trials of corticosteroids treatment for chronic inflammatory demyelinating polyradiculoneuropathy and enquiry from subject experts. SELECTION CRITERIA: Types of studies: All randomised or quasi-randomised trials Types of Participants: All patients with chronic inflammatory demyelinating polyradiculoneuropathy who were diagnosed by an internationally accepted definition. Types of interventions: Treatment with any form of corticosteroids or adrenocorticotropic hormone. Types of outcome measures: PRIMARY OUTCOME MEASURE: Change in disability 12 weeks after randomisation. SECONDARY OUTCOME MEASURES: 1. Change in impairment 12 weeks after randomisation. 2. Change in maximum motor nerve conduction velocity or compound muscle action potential amplitude after 12 weeks. 3. Side effects of corticosteroids. DATA COLLECTION AND ANALYSIS: One author extracted the data and the other checked them. MAIN RESULTS: We identified one randomised controlled trial, an open study in which 19 corticosteroids treated patients showed more improvement in impairment than 16 untreated controls after 12 weeks. Experience from large non-randomised studies suggests that steroids are beneficial. REVIEWER'S CONCLUSIONS: A single randomised controlled trial with 35 participants provided weak evidence to support the conclusion from non-randomised studies that oral corticosteroids reduce impairment in chronic inflammatory demyelinating polyradiculoneuropathy. Corticosteroids are known to have serious long term side effects. The long term risk and benefits have not been adequately studied. PMID- 11869621 TI - Surgery versus radiotherapy for muscle invasive bladder cancer. AB - BACKGROUND: Muscle invasive bladder cancer is a serious clinical problem and is fatal for the majority of patients. Alternative treatments for this condition are radical cystectomy or radical radiotherapy. The choice of treatment varies according to the resident country. The ideal treatment would be a bladder preserving therapy with total eradication of the tumour without compromising survival. OBJECTIVES: The objective of this review was to compare the overall survival after radical surgery (cystectomy) versus radical radiotherapy in patients with muscle invasive cancer. SEARCH STRATEGY: We searched the Cochrane Controlled Trials Register (July 2001), MEDLINE (July 2001), EMBASE (July 2001), CancerLIT (July 2001), Healthstar (July 2001) and the Database of Abstracts of Reviews of Effectiveness (July 2001). Attempts to contact authors of unpublished data were undertaken. SELECTION CRITERIA: Randomised trials comparing surgery versus radiotherapy were eligible for assessment. DATA COLLECTION AND ANALYSIS: Three reviewers assessed trial quality based on the Cochrane Guidelines. Data were extracted from the text of the article or extrapolated from the Kaplan-Meier plot. The Peto odds ratio was determined to compare the overall survival and disease-specific survival. Analysis was performed on an intention-to-treat basis and treatment actually received. MAIN RESULTS: Three randomised trials comparing pre-operative radiotherapy followed by radical cystectomy (surgery) versus radical radiotherapy with salvage cystectomy (radical radiotherapy) were eligible for assessment. These trials represented a total of 439 patients, 221 randomised to surgery and 218 to radical radiotherapy. Three trials were combined for the overall survival results and one for the disease-specific analysis [Bloom 1982]. The mean overall survival (intention-to-treat analysis) at 3 and 5 years were 45% and 36% for surgery, and 28% and 20% for radiotherapy, respectively. Peto odds ratio (95% Confidence Interval) analysis consistently favoured surgery in terms of overall survival. The results were significantly in favour of surgery at 3 years (OR = 1.91, 95% CI 1.30 -2.82) and at 5 years (OR = 1.85 95% CI 1.22 - 2.82). On a 'treatment received' basis, the results were significantly in favour of surgery at 3 (OR = 1.84, 95% CI 1.17 - 2.90) and 5 years (OR = 2.17, 95% CI 1.39 - 3.38) for overall survival and at 3 years (OR = 1.96, 95% CI 1.06,3.65) for disease-specific survival. REVIEWER'S CONCLUSIONS: The analysis of this review suggests that there is an overall survival benefit with radical surgery compared to radical radiotherapy in patients with muscle-invasive bladder cancer. However, it must be considered that only three trials were included for analysis, the patients numbers were small and that many patients did not receive the treatment they were randomised to. It must also be noted that many improvements in both radiotherapy and surgery have taken place since the initiation of these trials. PMID- 11869622 TI - Curative surgery for obstruction from primary left colorectal carcinoma: primary or staged resection? AB - BACKGROUND: In 8 to 29% of patients with colorectal carcinoma, obstruction is the main symptom at diagnosis, and 85% of patients undergoing emergency colorectal surgery have obstruction from colorectal carcinoma. The prognosis of patients who undergo emergency surgery for obstruction is often poor. So far, two types of surgical approach have been used for this condition: primary resection (primary anastomosis or Hartmann's procedure) with simultaneous treatment of carcinoma and obstruction or staged resection (treatment of the obstruction prior to resection). However, neither strategy has been found to have any advantages over the other. OBJECTIVES: To ascertain whether primary resection in patients with obstruction from left colorectal carcinoma has advantages over staged resection in terms of morbidity and mortality. SEARCH STRATEGY: Electronic database searches of Cochrane Controlled Trials Register, Medline, Cancerlit, Embase. Hand searching of the most important journals in the fields of oncology and surgery from the year 2000 until the time of writing. SELECTION CRITERIA: Randomised Clinical Trials (RCT) and Controlled Clinical Trials (CCT), in which a group of patients who undergo primary resection for intestinal obstruction from left primary colorectal carcinoma is compared with a group of patients who undergo staged resection for the same condition. Since only one study of this type was available, we considered all other studies, except for case-controls, on the basis of the best possible available evidence. Studies written in English, French, German and Italian and, where necessary, other languages, were considered. DATA COLLECTION AND ANALYSIS: Two reviewers (GLDS, CG) examined all the citations and abstracts derived from the electronic search strategy. Reports of potentially relevant trials were retrieved in full. Both reviewers independently applied the selection criteria to trials reports. Reviewers were not blind to the names of institutions, journals or authors of trials. A third opinion (SP, ML) was obtained to resolve disagreements. MAIN RESULTS: The electronic database searches yielded a total of 1412 citations: Medline 1026, Embase 386. From these, one study for potential inclusion was identified, but was then excluded. REVIEWER'S CONCLUSIONS: The limited number of identified trials together with their methodological weaknesses do not allow a reliable assessment of the role of either therapeutic strategy in the treatment of patients with bowel obstruction from colorectal carcinoma. It would appear advisable to conduct high quality large scale RCT to establish which treatment is more effective. However, it is doubtful whether they could be carried out in a timely and satisfactory way in this particular surgical context. PMID- 11869623 TI - Weighted vaginal cones for urinary incontinence. AB - BACKGROUND: Pelvic floor muscle training has long been the most common form of conservative treatment for stress urinary incontinence. Weighted vaginal cones can be used to help women to train their pelvic floor muscles. Cones are inserted into the vagina and the pelvic floor is contracted to prevent them slipping out. OBJECTIVES: To evaluate the effects of weighted vaginal cones in the treatment of women with urinary incontinence. SEARCH STRATEGY: We searched the Cochrane Incontinence Group specialised register (to February 2001), MEDLINE (January 1966 to August 2001), EMBASE (January 1988 to August 2001) and reference lists of relevant articles. SELECTION CRITERIA: Randomised or quasi-randomised controlled trials comparing weighted vaginal cones with alternative treatments or no treatment in women with urinary incontinence. DATA COLLECTION AND ANALYSIS: Three reviewers independently assessed studies for inclusion and trial quality. Data was extracted by one reviewer and cross checked by the others. Study authors were contacted for extra information. MAIN RESULTS: Fifteen studies, involving 1126 women of whom 466 received cones, were included. All of the trials were small and in many the quality was hard to judge. Outcome measures differed between studies, making the results difficult to combine. Some studies reported high drop out rates with both cone and comparison treatments. Four of the studies recruited women with symptoms of stress incontinence without urodynamic confirmation. Six trials were only published as abstracts. Cones were better than no active treatment (RR for failure to cure incontinence 0.74, 95% CI 0.59 to 0.93). There was little evidence of difference between cones and PFMT (RR 1.09, 95% CI 0.86 to 1.38) or electrostimulation (RR 1, 95% CI 0.89 to 1.13), but the confidence intervals were wide. There was not enough evidence to show that that cones plus PFMT was different to either cones alone or PFMT alone. Only two studies used a Quality of Life measure and no study looked at economic outcomes. REVIEWER'S CONCLUSIONS: This review provides some evidence that weighted vaginal cones are better than no active treatment in women with stress urinary incontinence and may be of similar effectiveness to PFMT and electrostimulation. This conclusion must remain tentative until further larger high quality studies are carried out using comparable and relevant outcome measures. Some women treated with cones, pelvic floor muscle training or electrostimulation drop out of treatment early. Therefore, cones should be offered as one option so that if women find them unacceptable they know there are other treatments available. PMID- 11869624 TI - Transcutaneous electrical nerve stimulation and acupuncture for primary dysmenorrhoea. AB - BACKGROUND: Dysmenorrhoea is the occurrence of painful menstrual cramps of the uterus. Medical therapy for dysmenorrhoea commonly consists of nonsteroidal anti inflammatory drugs or the oral contraceptive pill both of which work by reducing myometrial (uterine muscle) activity. However, these treatments are accompanied by a number of side effects, making an effective non-pharmacological method of treating dysmenorrhoea of potential value. Transcutaneous electrical nerve stimulation (TENS) is a treatment that has been shown to be effective for pain relief in a variety of conditions. Electrodes are placed on the skin and electric current applied at different pulse rates (frequencies) and intensities is used to stimulate these areas so as to provide pain relief. In dysmenorrhoea. TENS is thought to work by alteration of the body's ability to receive or perceive pain signals rather than by having a direct effect on the uterine contractions. Acupuncture may also be indicated as a useful, non-pharmacological method for treating dysmenorrhoea. Acupuncture is thought to excite receptors or nerve fibres which, through a complicated interaction with mediators such as serotonin and endorphins, blocks pain impulses. Acupuncture typically involves penetration of the skin by fine, solid metallic needles, which are manipulated manually or by electrical stimulation. OBJECTIVES: To determine the effectiveness of high and low frequency transcutaneous electrical nerve stimulation and acupuncture when compared to each other, placebo, no treatment, or medical treatment for primary dysmenorrhoea. SEARCH STRATEGY: Electronic searches of the Cochrane Menstrual Disorders and Subfertility Group Register of controlled trials, CCTR (Cochrane Library Issue 3, 2001), MEDLINE, EMBASE, CINAHL, Bio extracts, PsycLIT and SPORTDiscus were performed in August 2001 to identify relevant randomised controlled trials (RCTs). The Cochrane Complementary Medicine Field's Register of controlled trials (CISCOM) was also searched. Attempts were also made to identify trials from the UK National Research Register, the Clinical Trial Register and the citation lists of review articles and included trials. In most cases, the first or corresponding author of each included trial was contacted for additional information. SELECTION CRITERIA: The inclusion criteria were randomised controlled trials of transcutaneous electrical nerve stimulation and acupuncture that compared these treatments to each other, placebo, no treatment, or medical treatment for primary dysmenorrhoea. Exclusion criteria were: mild, infrequent or secondary dysmenorrhoea and dysmenorrhoea associated with an IUD. DATA COLLECTION AND ANALYSIS: Nine RCTs were identified that fulfilled the inclusion criteria for this review, seven involving TENS, one acupuncture, and one both treatments. Quality assessment and data extraction were performed independently by two reviewers. Meta analysis was performed using odds ratios for dichotomous outcomes and weighted mean differences for continuous outcomes. Data unsuitable for meta analysis was reported as descriptive data and was also included for discussion. The outcome measures were pain relief (dichotomous, visual analogue scales, descriptive), adverse effects, use of analgesics additional to treatment and absence from work or school. MAIN RESULTS: Overall high frequency TENS was shown to be more effective for pain relief than placebo TENS. Low frequency TENS was found to be no more effective in reducing pain than placebo TENS. There were conflicting results regarding whether high frequency TENS is more effective than low frequency TENS. One small trial showed acupuncture to be significantly more effective for pain relief than both placebo acupuncture and two no treatment control groups. REVIEWER'S CONCLUSIONS: High frequency TENS was found to be effective for the treatment of dysmenorrhoea by a number of small trials. The minor adverse effects reported in one trial requires further investigation. There is insufficient evidence to determine the effectiveness of low frequency TENS in reducing dysmenorrhoea. There is also insufficient evidence to determine the effectiveness of acupuncture in reducing dysmenorrhoea, however a single small but methodologically sound trial of acupuncture suggests benefit for this modality. PMID- 11869625 TI - Pressurised metered dose inhalers versus all other hand-held inhaler devices to deliver beta-2 agonist bronchodilators for non-acute asthma. AB - BACKGROUND: A number of different inhaler devices are available to deliver beta2 agonist bronchodilators in asthma. These include hydrofluoroalkane (HFA) or chlorofluorocarbon (CFC)-free propelled pressurised metered dose inhalers (pMDIs) and dry powder devices. OBJECTIVES: To determine the clinical effectiveness of pMDI compared with any other available handheld inhaler device for the delivery of short-acting beta-2 agonist bronchodilators in non-acute asthma in children and adults. SEARCH STRATEGY: The Cochrane Collaboration Clinical Trials register was searched for studies as well as separate additional searches carried out on MEDLINE, EMBASE, CINAHL and also on the Current Contents Index as well as the Science Citation Index. In addition, 17 individual online respiratory journals and 12 electronically available clinical trial databases were also searched. The UK pharmaceutical companies who manufacture inhaled asthma medication were contacted in order to obtain details of any published or unpublished studies. SELECTION CRITERIA: - The full texts of all potentially relevant articles were reviewed independently by two reviewers. DATA COLLECTION AND ANALYSIS: Fixed and random effect models were used. Dichotomous outcomes were assessed using Odds Ratios or Relative Risks (RR) with 95% Confidence Intervals (95%CI). MAIN RESULTS: Eighty-four randomised controlled trials were included in this review, but few could be combined to assess a specific outcome for a given delivery device comparison. Only two studies required demonstration of adequate pMDI technique as an entry requirement. There were no difference between a standard CFC containing pMDI and any other device for most outcomes. Regular use of HFA pMDI containing salbutamol reduced the requirement for short courses of oral corticosteroids (3 trials, 519 patients: RR 0.67; 95% CI 0.49, 0.91); however the total number of exacerbations were unchanged (3 trials, 1271 patients: RR 1.0; 95% CI 0.75, 1.33). REVIEWER'S CONCLUSIONS: In patients with stable asthma, short acting beta-2 bronchodilators in standard CFC-pMDI's are as effective as any other devices. The effect of HFA-pMDI on requirement for oral corticosteroid courses to treat acute exacerbations should be confirmed. Effectiveness studies that use an intention-to-treat analysis are required. PMID- 11869626 TI - Pneumococcal vaccine for asthma. AB - BACKGROUND: Infection with Streptococcus pneumoniae is an important cause of pneumonia and other serious illnesses, particularly amongst those with certain high-risk medical conditions such as asthma. Although pneumococcal vaccine is routinely advocated for people with asthma, there is uncertainty about the evidence base that underpins this recommendation. OBJECTIVES: To determine the efficacy of pneumococcal vaccine in reducing mortality or morbidity from pneumococcal disease in asthmatics. SEARCH STRATEGY: Randomised controlled trials were identified using the Cochrane Airways Group's register derived from MEDLINE, EMBASE, and CINAHL electronic databases and hand searched respiratory journals and meeting abstracts. SELECTION CRITERIA: Randomised controlled trials, with or without blinding, in which pneumococcal vaccine has been compared with placebo or no treatment in people with clinician diagnosed asthma. DATA COLLECTION AND ANALYSIS: Two reviewers independently reviewed all abstracts and full papers of all articles of potential relevance were retrieved. Methodological quality was rated using the Cochrane approach and the Jadad rating scale. Data extraction was performed by one reviewer and checked independently by a second. We planned to perform quantitative analyses of outcomes on an intention-to-treat basis, where possible. MAIN RESULTS: Of the three papers retrieved, only one satisfied the inclusion criteria and the methodological quality of this study was low (unblinded and inadequate allocation concealment). None of the data could be aggregated in a meta-analysis. Comparisons in a sub-set of 30 asthmatic children prone to recurrent episodes of otitis media, showed that pneumococcal vaccination decreased the incidence of acute asthma exacerbations from 10 to 7 (per child per year). A further search conducted in September 2001 did not yield any further studies. REVIEWER'S CONCLUSIONS: This review found very limited evidence to support the routine use of pneumococcal vaccine in people with asthma. A randomised trial of vaccine efficacy in children and adults with asthma is needed. PMID- 11869627 TI - Pressurised metered-dose inhalers versus all other hand-held inhalers devices to deliver bronchodilators for chronic obstructive pulmonary disease. AB - BACKGROUND: Bronchodilator therapy for COPD may be delivered by a number of different inhaler devices. OBJECTIVES: To determine the efficacy of pressurised metered dose inhalers (pMDI) compared to any other handheld inhaler device for the delivery of bronchodilators in non-acute COPD. SEARCH STRATEGY: The Cochrane Collaboration, Asthma and Wheeze Randomised Controlled Clinical Trials register was searched for studies. The UK pharmaceutical companies who manufacture inhaled COPD medication were also contacted. SELECTION CRITERIA: Two reviewers independently reviewed the results of computerised search and any potentially relevant articles were obtained in full. DATA COLLECTION AND ANALYSIS: One reviewer extracted details of each trial and a second reviewer checked all extracted data. Dichotomous outcomes such as exacerbation rate were assessed using relative risk, with 95% confidence interval (CI). MAIN RESULTS: Fourteen studies appeared potentially relevant but only three studies (61 patients) met the entry criteria. Two studies compared a dry powder device (Turbuhaler or Rotahaler) with a pMDI for beta2-agonist delivery, and one (36 patients cross over design) the Respimat (soft mist device for ipratropium) vs a pMDI. For the Turbuhaler and Rotahaler, none of the reported outcome measures were significantly different. The Rotahaler study used a high and low dose of medication with or without large volume spacer. The study using the Respimat showed significant increases in FEV1 when compared to a pMDI (difference in change from base line 70 ml, 95% CI 10, 130 ml). The effect on change in FVC was of similar size. There were no differences between these two devices for any other reported outcomes. Although none of the included studies required prior patient ability to use any of the inhalers (and no study mentioned device training), it was assumed that all patients randomised into the study would have undergone training in use of the study inhalers and were capable of using those devices. REVIEWER'S CONCLUSIONS: In patients with stable COPD, pMDI produced similar outcomes to a dry powder device for delivering beta2-agonists, but the very small number of studies and included patients does not permit firm conclusions to be drawn. The soft mist device for ipratropium was more effective than a pMDI, but the data come from one small study. There need to be further well designed randomised controlled trials to define the role of inhaler devices using bronchodilators in stable COPD. PMID- 11869628 TI - Operative procedures for fissure in ano. AB - BACKGROUND: Operative techniques commonly used for fissure in ano include: anal stretch, open lateral sphincterotomy, closed lateral sphincterotomy, posterior midline sphincterotomy and to a lesser extent dermal flap coverage of the fissure. Reports of direct comparisons between operative techniques for anal fissure are variable in their results. These reports are either subject to selection bias (in non-randomized studies) or observer bias (in all studies) or have inadequate numbers of patients enrolled to answer the question of efficacy. OBJECTIVES: To determine the best technique for fissure surgery. SEARCH STRATEGY: The Cochrane Controlled Trials Register [Cochrane2000] and MEDLINE (1965-2000) were searched. The list of cited references in all included reports and several study authors also were helpful in finding additional comparative studies. SELECTION CRITERIA: All reports in which there was a direct comparison between at least two operative techniques were reviewed and when more than one report existed for any given pair, that report was included. If crude data were not presented in the report, the authors were contacted and crude data obtained. DATA COLLECTION AND ANALYSIS: The two most commonly used end points in all reported studies were persistence of the fissure and post operative incontinence of flatus. These are the only two endpoints included in the meta-analysis. MAIN RESULTS: Anal stretch has a higher risk of fissure persistence than internal sphincterotomy and also a significantly higher risk of minor incontinence than sphincterotomy. The combined results of open versus closed partial lateral internal sphincterotomy show little difference between the two procedures both in fissure persistence and risk of incontinence. REVIEWER'S CONCLUSIONS: Anal stretch and posterior midline internal sphincterotomy should probably be abandoned in the treatment of chronic anal fissure in adults. For those patients requiring surgery for anal fissure, open and closed partial lateral internal sphincterotomy appear to be equally efficacious. More data are needed to assess the effectiveness of posterior internal sphincterotomy. PMID- 11869629 TI - Follow-up strategies for patients treated for non-metastatic colorectal cancer. AB - BACKGROUND: It is common clinical practise to follow patients with colorectal cancer (CRC) for several years following their definitive surgery and/or adjuvant therapy. Despite this widespread practice there is considerable controversy about how often patients should be seen, what tests should be performed and whether these varying strategies have any significant impact on patient outcomes. OBJECTIVES: To review the available evidence concerning the benefits of intensive follow-up of colorectal cancer patients with respect to survival. Secondary endpoints include time to diagnosis of recurrence, quality of life and the harms and costs of surveillance and investigations. SEARCH STRATEGY: Relevant trials were identified by electronic searches of MEDLINE, EMBASE, CINAHL, CANCERLIT, Cochrane Controlled Trials Register, Science Citation Index, conference proceedings, trial registers, reference lists and contact with experts in the field. SELECTION CRITERIA: Only randomised controlled trials comparing different follow-up strategies for patients with non-metastatic CRC treated with curative intent were included. DATA COLLECTION AND ANALYSIS: Trial eligibility and methodological quality were assessed independently by the three reviewers. MAIN RESULTS: Five trials were included. There was evidence that an overall survival benefit at 5 years exists for patients undergoing more intensive follow-up (OR = 0.67, 95% confidence interval 0.53 - 0.84; RD = -0.07, CI -0.12 - -0.02). The absolute number of recurrences was similar (OR = 0.91; 95% confidence interval 0.72 - 1.14; RD = 0.00, CI -0.07 - 0.07) and although the weighted mean difference for the time to recurrence was significantly reduced by 6.75 (95% confidence interval -11.06 - -2.44) there was significant heterogeneity between the studies. Analyses demonstrated a mortality benefit for performing more tests versus fewer tests (OR = 0.66; 95% confidence interval 0.46 - 0.95) and liver imaging versus no liver imaging (OR = 0.66; 95% confidence interval 0.46 - 0.95). However when both these results are expressed as a risk difference this significance is lost (RD = -0.06; CI -0.25 - 0.13). No useful data on quality of life, harms or cost-effectiveness were available for further analysis. REVIEWER'S CONCLUSIONS: The results of our review suggest that there is an overall survival benefit for intensifying the follow-up of patients after curative surgery for colorectal cancer. Because of the wide variation in the follow-up programmes used in the included studies it is not possible to infer from the data the best combination and frequency of clinic (or family practice) visits, blood tests, endoscopic procedures and radiological investigations to maximise the outcomes for these patients. Nor is it possible to estimate the potential harms or costs of intensifying follow-up for these patients in order to adopt a cost-effective approach in this clinical area. Large clinical trials underway or about to commence are likely to contribute valuable further information to clarify these areas of clinical uncertainty. PMID- 11869630 TI - Antibiotics versus control for toxoplasma retinochoroiditis. AB - BACKGROUND: Acute toxoplasma retinochoroiditis causes transient symptoms of ocular discomfort and may lead to permanent visual loss. Antibiotic treatment primarily aims to reduce the risk of permanent visual loss, recurrent retinochoroiditis, and the severity and duration of acute symptoms. There is uncertainty about the effectiveness of antibiotic treatment. OBJECTIVES: The objective of this review was to compare the effects of antibiotics versus placebo or no treatment for toxoplasma retinochoroiditis. SEARCH STRATEGY: We searched the Cochrane Controlled Trials Register - CENTRAL/CCTR, which contains the Cochrane Eyes and Vision Group Specialised Register (Cochrane Library Issue 2, 2001), MEDLINE (1966 to August 2001), EMBASE (1980 to September 2001), Dissertation Abstracts (1861 to June 2001), LILACS (1982 to 1998), Pascal (1984 to March 2000), proceedings of the Association for Research in Vision and Ophthalmology (1980 to 2001), international symposia on uveitis, and reference lists of review articles. Pharmaceutical companies were contacted for unpublished trials. SELECTION CRITERIA: We included randomised controlled trials that compared any systemic antibiotic treatment against placebo or no treatment. Trials that included immunocompromised patients were excluded. DATA COLLECTION AND ANALYSIS: The primary outcomes for this review were visual acuity at least three months after treatment and risk of recurrent retinochoroiditis. Secondary outcomes were improvement in symptoms and signs of intraocular inflammation, size of lesion and adverse events. Effect measures were pooled using a random effects model. MAIN RESULTS: Three trials, which randomised a total of 173 participants, met the inclusion criteria. All trials were methodologically poor. None reported the effect of treatment on visual acuity. Two studies reported results for recurrent retinochoroiditis: one (124 participants) found a significant reduction in participants with chronic recurrent disease who were treated for 14 months: relative risk 0.28 (95% confidence interval 0.10 to 0.78); the other (20 participants) found no evidence of an effect in participants with acute toxoplasma retinochoroiditis (relative risk 1.00, 95% confidence interval 0.07 to 13.87). Two studies reported an improvement in intraocular inflammation in treated compared with untreated participants and one study reported no difference. Two studies found an increased risk of adverse events in treated participants. REVIEWER'S CONCLUSIONS: There is a lack of evidence to support routine antibiotic treatment for acute toxoplasma retinochoroiditis. There is weak evidence to suggest that long-term treatment of patients with chronic recurrent toxoplasma retinochoroiditis may reduce recurrence. Placebo controlled trials of patients with acute and chronic toxoplasma retinochoroiditis affecting any part of the retina are required to determine the effectiveness of antibiotic treatment. PMID- 11869631 TI - Interventions for involutional lower lid entropion. AB - BACKGROUND: Entropion is a condition in which the eyelid margin turns in against the eyeball. The involutional or senile type of entropion is one of the most common lower lid malpositions in the elderly. The interventions described and currently used for the treatment of this condition are surgical in nature, although non-surgical temporary medical treatment for the early stages of entropion has also been reported. The relative effectiveness of these interventions has not yet been resolved. OBJECTIVES: The purpose of this review is to examine the effect of interventions for involutional entropion and to assess whether any method is superior to the others. SEARCH STRATEGY: We searched the Cochrane Controlled Trials Register - CENTRAL/CCTR, which contains the Cochrane Eyes and Vision Group specialised register (Cochrane Library Issue 3, 2000), MEDLINE (1966 to October 2000), EMBASE (1980 to September 2000), the metaRegister of Controlled Trials (mRCT), oculoplastic textbooks, conference proceedings and bibliographies of relevant reports. We contacted investigators and experts in the field for details of other published and unpublished studies. SELECTION CRITERIA: The criteria for including studies in this review were randomised controlled trials where one intervention for involutional lower lid entropion has been compared to another method of treatment in people older than 60 years of age with involutional lower lid entropion. DATA COLLECTION AND ANALYSIS: Both reviewers assessed the search results according to the criteria for including studies in the review. No studies were found that met these criteria, and therefore no data were collected and no meta-analysis was performed. MAIN RESULTS: Since no studies met our selection criteria no results were produced. REVIEWER'S CONCLUSIONS: No conclusions could be reached about the effectiveness of interventions for involutional lower lid entropion, although retrospective case series studies suggest that some forms of treatment may be associated with higher recurrence rates than others. Lack of suitable data highlights the need for randomised controlled trials in the fields of entropion management. PMID- 11869632 TI - Emergency sclerotherapy versus medical interventions for bleeding oesophageal varices in cirrhotic patients. AB - BACKGROUND: Emergency sclerotherapy is widely used as a first line therapy for variceal bleeding in cirrhosis, although pharmacological treatment may stop bleeding in the majority of patients. OBJECTIVES: To assess whether emergency sclerotherapy is superior to pharmacological treatment for variceal bleeding in cirrhosis. SEARCH STRATEGY: Electronic and manual searches were combined until April 2001. SELECTION CRITERIA: Randomised clinical trials comparing sclerotherapy with vasoactive treatments (vasopressin (plus minus nitroglycerin), terlipressin, somatostatin, or octreotide) for acute variceal bleeding in cirrhotic patients. DATA COLLECTION AND ANALYSIS: Two independent reviewers identified eligible trials and extracted data. Outcome measures were failure to control bleeding, five-day treatment failure, rebleeding before other elective treatments, 42-day rebleeding, mortality before other elective treatments, 42-day mortality, number of blood transfusions, and adverse events. Data were analysed by a random effects model according to the vasoactive treatment. Sensitivity analyses included combined analysis of all the trials irrespective of the vasoactive drug, fixed effects model analyses, type of publication, methodological quality, and adequacy of generation of the randomisation list and of allocation concealment. MAIN RESULTS: Twelve trials including 1146 patients (pts) were identified. One trial compared sclerotherapy with vasopressin, one with terlipressin, four with somatostatin, and six with octreotide. No significant differences were found comparing sclerotherapy with each vasoactive drug for any outcomes. Combining all the trials irrespective of the vasoactive drug, risk differences (95% confidence intervals) were: failure to control bleeding (11 RCTs, 977 pts) -0.03 (-0.07 to 0.01); five-day failure rate (7 RCTs, 759 pts) -0.05 (-0.12 to 0.01); rebleeding (11 RCTs, 1082 pts) -0.01(-0.06 to 0.04); rebleeding before other elective treatments (9 RCTs, 975 pts) -0.02 (-0.06 to 0.03); mortality (12 RCTs, 1146 pts) -0.04 (-0.08 to 0.00); mortality before other elective treatments (5 RCTs, 474 pts) -0.02 (-0.07 to 0.04); transfused blood units (7 RCTs, 793 pts) (weighted mean difference) -0.17 (-0.52 to 0.19). Adverse events (11 RCTs, 1082 pts) and serious adverse events (5 RCTs, 602 pts) were significantly more frequent with sclerotherapy: risk differences 0.08 (0.02 to 0.14) and 0.05 (0.02 to 0.08), respectively. Results were consistent across all the other sensitivity analyses. REVIEWER'S CONCLUSIONS: We found no convincing evidence to support the use of emergency sclerotherapy for variceal bleeding in cirrhosis as the first, single treatment when compared with vasoactive drugs. PMID- 11869633 TI - Ribavirin with or without alpha interferon versus no intervention, placebo or alpha interferon for chronic hepatitis C. AB - BACKGROUND: Hepatitis C is a major cause of liver-related morbidity and mortality. Ribavirin plus interferon combination therapy is presently considered the optimal treatment of interferon naive patients with chronic hepatitis C, but its role in relapsers and non-responders to previous interferon therapy is not established. OBJECTIVES: To assess the efficacy and safety of ribavirin alone or in combination with alpha interferon in interferon naive patients, relapsers, and non-responders with chronic hepatitis C. SEARCH STRATEGY: Eligible trials were identified through electronic databases, manual searches of bibliographies and journals, authors of trials, and pharmaceutical companies producing ribavirin or interferon (August 2001). SELECTION CRITERIA: We included all randomised trials comparing ribavirin with or without alpha interferon versus no intervention, placebo, or alpha interferon for chronic hepatitis C. DATA COLLECTION AND ANALYSIS: The primary outcome measures were the 'sustained' (six months after treatment) virological response, and morbidity plus mortality. The secondary outcome measures were the 'end of treatment' and 'sustained' biochemical response, the 'end of treatment' virologic response, histology, quality of life, and adverse events. MAIN RESULTS: We included eight trials in which 271 patients were randomised to ribavirin versus placebo or no intervention and 48 trials in which 6585 patients were randomised to interferon with or without ribavirin. Compared with placebo or no intervention, ribavirin monotherapy had no significant effect on the virological response or histology and only a transient effect on the biochemical response. Compared with interferon, combination therapy reduced the risk of not having a sustained virological response by 26% in naive patients (relative risk (RR) 0.74; 95% confidence interval (CI) 0.70-0.78), 33% in relapsers (RR 0.67; 95% CI 0.57-0.78), and 11% in non-responders (RR 0.89; 95% CI 0.83-0.96). There was no significant effect on morbidity plus mortality (Peto odds ratio 0.45; 95% CI 0.19-1.06). Irrespective of previous therapy, combination therapy significantly reduced the risk of not having a sustained biochemical response (RR 0.76; 95% CI 0.59-0.84) or improved histology (RR 0.67; 95% CI 0.56 0.81). Combination therapy also significantly increased the risk of treatment discontinuation (RR 1.28; 95% CI 1.07-1.52) and several types of adverse events. REVIEWER'S CONCLUSIONS: Combination therapy increased the number of naive patients, relapsers, and non-responders with a sustained virological, biochemical, or histological response, but also the occurrence of adverse events. PMID- 11869634 TI - Laparoscopic colposuspension for urinary incontinence in women. AB - BACKGROUND: Laparoscopic colposuspension is a relatively new operation for the treatment of women with stress urinary incontinence with the presumed advantages over traditional Burch colposuspension of avoiding major incisions, shorter hospital stay, and quicker return to normal activities. A variety of approaches and methods are used. OBJECTIVES: To determine the effects of laparoscopic colposuspension surgery on urinary incontinence. SEARCH STRATEGY: We searched the Cochrane Incontinence Group specialised register. The date of the most recent search was April 2001. Additional trials were sought from other sources such as reference lists, conference proceedings, reviews and unpublished research. SELECTION CRITERIA: Randomised or quasi-randomised controlled trials in women with symptomatic or urodynamic diagnosis of stress or mixed incontinence that included laparoscopic surgery in at least one arm of the study. DATA COLLECTION AND ANALYSIS: Trials were evaluated for methodological quality and appropriateness for inclusion by the reviewers. Data were extracted by two of the reviewers and cross checked by another. Trial data were analysed by intervention. Where appropriate, a summary statistic was calculated. MAIN RESULTS: Eight eligible trials were identified. Five included 233 women receiving a laparoscopic and 254 women an open colposuspension. Whilst the women's subjective impression of cure seemed similar for both procedures up to 18 months there was some evidence of poorer results on objective outcomes. A single trial suggested poorer long-term performance, but this may reflect surgical inexperience of laparoscopic colposuspension. No significant differences were observed for post-operative urgency, voiding dysfunction or de novo detrusor instability. Trends were shown towards a higher complication rate, longer operating time, less intraoperative blood loss, less postoperative pain, shorter hospital stay, quicker return to normal activities, and shorter duration of catheterisation for laparoscopic compared with open colposuspension. Significantly higher subjective and objective (dry on 'ultrashort' pad test) one year cure rates were found for women randomised to two paravaginal sutures compared with one suture in a single trial (89% vs 65% and 83% vs 58% respectively). One study compared sutures with mesh and staples for laparoscopic colposuspension but it was too small to allow a reliable comparison. One study compared transperitoneal with extraperitoneal access for laparoscopic colposuspension but it was also small and of poor quality. REVIEWER'S CONCLUSIONS: The long-term performance of laparoscopic colposuspension is uncertain. Currently available evidence suggests that it may be poorer than open colposuspension. Like other laparoscopically performed operations, laparoscopic colposuspension leads to a quicker recovery, but takes longer to perform and may be associated with more surgical complications. If it is performed, two paravaginal sutures appear to be more effective than one. The place of laparoscopic colposuspension in clinical practice should become clearer when ongoing trials are reported and when there are more data available describing long-term cure results. PMID- 11869635 TI - Nasal intermittent positive pressure ventilation (NIPPV) versus nasal continuous positive airway pressure (NCPAP) for apnea of prematurity. AB - BACKGROUND: Apnea of prematurity is almost universal in infants who are born before 34 weeks gestation. Previous randomised trials and systematic reviews have found methylxanthines to be effective in preventing apnea of prematurity. However, recent concerns about potential long term side effects of methylxanthines on the neurodevelopment of low birth weight infants have led to an increased interest in alternate methods of treating apnea of prematurity. Nasal continuous positive airway pressure (NCPAP) is a useful method of respiratory support which reduces the incidence of obstructive or mixed apnea. However, apneic infants managed with NCPAP, with or without methylxanthines, sometimes require endotracheal intubation with its attendant morbidity and cost. Nasal intermittent positive pressure ventilation (NIPPV) is a simple, effective mode of respiratory support for older children and adults. It has been used to treat apnea in preterm infants but case reports of gastrointestinal perforations have limited its widespread use. OBJECTIVES: In preterm infants with recurrent apnea, does treatment with NIPPV lead to a greater reduction in apnea and need for intubation and mechanical ventilation, as compared with treatment with NCPAP? Does NIPPV increase the incidence of gastrointestinal complications, i.e. gastric distension leading to cessation of feeds, or perforation? SEARCH STRATEGY: MEDLINE was searched (1966-Oct week 2, 2001). Other sources included the Cochrane Controlled Trials Register (Cochrane Library, Disk Issue 3, 2001) and CINAHL (1982-Sept week 4, 2001). Also used were expert informants, previous reviews including cross-references, and conference and symposia proceedings. SELECTION CRITERIA: All randomised and quasi-randomised trials were included. Participants included unventilated preterm infants experiencing apnea of prematurity. Interventions compared were intermittent positive pressure ventilation administered via the nasal route, either by short nasal prongs or nasopharyngeal tube, and nasal CPAP delivered by the same methods. Types of outcome measures: - failure of therapy as defined by apnea that is frequent or severe requiring additional ventilatory support - rates of endotracheal intubation - rates of apnea and bradycardia expressed as events per hour - gastrointestinal complications i.e. abdominal distension requiring cessation of feeds, or GI perforation DATA COLLECTION AND ANALYSIS: Data were extracted independently by the three reviewers. The trials were analysed using relative risk (RR), risk difference (RD) and number needed to treat (NNT) for dichotomous data; means and weighted mean difference (WMD) were used for continuous data. MAIN RESULTS: Two trials, enrolling 54 infants in total, fulfilled the inclusion criteria. Both reported only the short term results (4 to 6 hours) of the interventions. Only one infant (randomised to NCPAP) required intubation during this period. Ryan (1989), in a cross over study of 20 infants, showed no significant difference in rates of apnea (events/hr) between the 2 interventions [WMD -0.10 (-0.53,0.33)]. Lin (1998) randomised 34 infants and demonstrated a greater reduction in frequency of apneas (events/hr) with NIPPV compared to NCPAP [WMD -1.19 (-2.31, 0.07)]. Meta-analysis of both trials showed no difference in pCO2 (mmHg) at the end of the 4-6 hour study period [WMD 0.95 (-3.05,4.94)]. No data were reported on gastrointestinal complications. REVIEWER'S CONCLUSIONS: IMPLICATIONS FOR PRACTICE: NIPPV may be a useful method of augmenting the beneficial effects of NCPAP in preterm infants with apnea that is frequent or severe. Its use appears to reduce the frequency of apneas more effectively than NCPAP. Additional safety and efficacy data are required before recommending NIPPV as standard therapy for apnea. IMPLICATIONS FOR RESEARCH: Future trials with sufficient power should assess the efficacy (reduction in failure of therapy) and safety (GI complications) of NIPPV. Outcomes should be assessed throughout the entire period during which the infant requires assisted ventilation. The recent ability to synchronise NIPPV with an infant's spontaneous respirations is a promising development requiring further assessment. PMID- 11869636 TI - Fluticasone versus beclomethasone or budesonide for chronic asthma. AB - BACKGROUND: Beclomethasone dipropionate (BDP) and budesonide (BUD) are commonly prescribed inhaled corticosteroids for the treatment of asthma, Fluticasone propionate (FP) is newer agent with greater potency in in-vitro assays. OBJECTIVES: To compare the efficacy and safety of Fluticasone to Beclomethasone or Budesonide in the treatment of chronic asthma. SEARCH STRATEGY: We searched the Cochrane Airways Group Trial Register (1999) and reference lists of articles. We contacted trialists and pharmaceutical companies for additional studies and searched abstracts of major respiratory society meetings (1997-1999). SELECTION CRITERIA: Randomised trials in children and adults comparing Fluticasone to either Beclomethasone or Budesonide in the treatment of chronic asthma. Two reviewers independently assessed articles for inclusion and methodological quality. DATA COLLECTION AND ANALYSIS: One reviewer extracted data. Quantitative analyses where undertaken using Review Manager 4.0.3 with Metaview 3.1. MAIN RESULTS: 42 studies (>10,000 patients) met the inclusion criteria. Methodological quality was variable. When compared at a FP:BUD/BDP dose ratio of 1:2, fluticasone produced a significantly greater FEV1 (Weighted Mean Difference (WMD) 0.11 litres, 95% Confidence Interval (CI) 0.01, 0.20 litres), morning PEF (WMD 13 L/min, 95%CI 5, 22 L/min) and evening PEF (WMD 11 L/min, 95%CI 1, 20 L/min). This applied to all drug doses, age groups, and delivery devices, although subgroup analyses suggested that the relative benefit of FP may be greater in more severe patients treated with higher doses of inhaled corticosteroid. No difference between fluticasone and beclomethasone or budesonide were seen for trial withdrawals (Peto OR 0.77, 95%CI 0.54, 1.10). Symptoms and rescue medication use were widely reported but few trials provided sufficient data for analysis. A higher likelihood of pharyngitis (Peto Odds Ratio 2.16; 95% CI 1.42, 3.28) was apparent when patients were treated with fluticasone at twice the dose of BDP/BUD, although was unexplained heterogeneity in this effect between trials. There was no difference in the likelihood of oral Candidiasis. Plasma cortisol and 24 hour urinary cortisol were measured frequently but data presentation was limited. REVIEWER'S CONCLUSIONS: Fluticasone given at half the daily dose of beclomethasone or budesonide leads to small improvements in measures of airway calibre, but it appears to have a higher risk of causing side-effects when given at the same daily dose. PMID- 11869637 TI - Thermotherapy for treating rheumatoid arthritis. AB - BACKGROUND: Thermotherapy is often used as adjunct in the treatment of rheumatoid arthritis (RA) by rehabilitation specialists. OBJECTIVES: To evaluate the effectiveness of different thermotherapy applications on objective and subjective measures of disease activity in patients with RA. SEARCH STRATEGY: We searched Medline, EMBASE, Pedro, Current Contents, Sports Discus and CINAHL up to and including September 2001. The Cochrane Field of Rehabilitation and related therapies and the Cochrane Musculoskeletal Review Group were also contacted for a search of their specialized registers. Hand searching was conducted on all retrieved articles for additional articles. SELECTION CRITERIA: Comparative controlled studies, such as randomized controlled trials, controlled clinical trials, cohort studies or case/control studies, of thermotherapy compared to control or active interventions in patients with RA were eligible. No language restrictions were applied. Abstracts were accepted. DATA COLLECTION AND ANALYSIS: Two independent reviewers identified potential articles from the literature search (VR, LB). These reviewers extracted data using pre-defined extraction forms. Consensus was reached on all data extraction. Quality was assessed by two reviewers using a 5 point scale that measured the quality of randomization, double-blinding and description of withdrawals. MAIN RESULTS: Seven studies (n=328 subjects) met the inclusion criteria. The results of this systematic review of thermotherapy for RA found that there was no significant effect of hot and ice packs applications (Ivey 1994), cryotherapy (Rembe 1970) and faradic baths (Hawkes 1986) on objective measures of disease activity including joint swelling, pain, medication intake, range of motion (ROM), grip strength, hand function compared to a control (no treatment) or active therapy. There is no significant difference between wax and therapeutic ultrasound as well as between wax and faradic bath combined to ultrasound for all the outcomes measured after 1, 2 or 3 week(s) of treatment (Hawkes 1986). There was no difference in patient preference for all types of thermotherapy. No harmful effects of thermotherapy were reported. REVIEWER'S CONCLUSIONS: Superficial moist heat and cryotherapy can be used as a palliative therapy. Paraffin wax baths combined with exercises can be recommended for beneficial short term effects for arthritic hands. These conclusions are limited by methodological considerations such as the poor quality of trials. PMID- 11869638 TI - Perazine for schizophrenia. AB - BACKGROUND: Perazine is an old phenothiazine derivative used for the treatment of people with schizophrenia which has a reputed low level of extrapyramidal side effects. However, its use is restricted in the sense that - to the best knowledge of the reviewers - it is only marketed in Germany, Poland, Yugoslavia and the Netherlands. OBJECTIVES: To examine the effects of perazine for those with schizophrenia, and schizophrenia-like psychoses. SEARCH STRATEGY: Electronic searches of the Cochrane Schizophrenia Group's register which includes relevant randomised controlled trials from the bibliographic databases Biological Abstracts, CINAHL, The Cochrane Library, EMBASE, MEDLINE, PsycLIT, LILACS, PSYNDEX, Sociological Abstracts and Sociofile were undertaken. References of all included studies were searched for further trials. Pharmaceutical companies and authors of trials were contacted. SELECTION CRITERIA: All randomised controlled trials that compared perazine with other treatments for people with schizophrenia and/or schizophrenia-like psychoses. DATA COLLECTION AND ANALYSIS: Citations and, where possible, abstracts were independently inspected by two reviewers, papers ordered, re-inspected and quality assessed. Data were independently extracted. Data were excluded if loss to follow up was greater than 50%. For homogeneous dichotomous data the Relative Risk (RR), 95% confidence interval (CI) and, where appropriate, the number needed to treat (NNT) were calculated on an intention-to treat basis. For continuous data, weighted mean differences were calculated (WMD). All data were inspected for heterogeneity. MAIN RESULTS: Six trials with a total of 288 participants are included. According to only one trial with 95 participants perazine appeared superior to active placebo (trimipramine) at five weeks for the outcome of 'no important global improvement' (n=95, RR 0.6, CI 0.3 0.9, NNT 4, CI 2-17), but there was no difference in various measures of mental state. The side-effect risk of perazine compared to placebo could not be estimated because they were not reported. Five small trials including only 193 participants which compared perazine with other antipsychotics were incompletely reported and the outcomes were presented in various ways so that meta-analysis was not possible in most occasions. A similar number of participants receiving perazine or comparator antipsychotics left the studies early (n=193, RR 0.9, CI 0.5-1.4). The results on efficacy were controversial and need further assessment by randomised controlled trials. No obvious differences in adverse events between perazine and other antipsychotics could be derived from these limited data. Two haloperidol comparisons did not present extrapyramidal side-effects in a way usable for meta-analysis, but three small comparisons with the atypical antipsychotics zotepine and amisulpride showed no higher risk of akathisia (n=111, RR 0.3, CI 0.1-1.1), dyskinesia (n=111, RR 0.5, CI 0.1-3.5), parkinsonism (n=81, RR 1.2, CI 0.5-2.8) or tremor (n=40, RR 0.8, CI 0.3-2.3) with perazine. REVIEWER'S CONCLUSIONS: The number, size and reporting of randomised controlled perazine trials is insufficient to present firm conclusions about the properties of this antipsychotic. It is possible that perazine is associated with a similar risk of extrapyramidal side-effects as some atypical antipsychotics, and this should be clarified in larger, well-designed trials. PMID- 11869639 TI - Reduced osmolarity oral rehydration solution for treating dehydration caused by acute diarrhoea in children. AB - BACKGROUND: Oral rehydration solution (ORS) has reduced childhood deaths from diarrhoea in many countries. Recent studies suggest that the currently recommended formulation of ORS recommended by the World Health Organization (WHO) may not be optimal, and solutions that contain lower concentrations of sodium and glucose may be more effective. OBJECTIVES: To compare reduced osmolarity ORS with WHO standard ORS in children with acute diarrhoea. SEARCH STRATEGY: The Cochrane Collaboration Trials Register (on The Cochrane Library, Issue 2, 2001), MEDLINE (1966 to June 2001), EMBASE (1988 to May 2001), and Current Contents (June 2001) were searched. Additional trials were identified by hand searching. Content experts were contacted. SELECTION CRITERIA: Randomized controlled trials comparing reduced osmolarity ORS with the WHO standard ORS formulation. The primary outcome was unscheduled intravenous fluid infusion. Secondary outcomes were measures of clinical illness. DATA COLLECTION AND ANALYSIS: Two reviewers extracted data. We tested for heterogeneity using the Chi-square statistic, conducted sensitivity analysis by allocation concealment, and the regression approach to assess funnel plot asymmetry from selective trial publication. MAIN RESULTS: The primary outcome, unscheduled intravenous fluid infusion, was reported in 11 trials. In a meta-analysis of 8 trials, reduced osmolarity ORS was associated with fewer unscheduled intravenous fluid infusions compared with WHO standard ORS (Mantel Haenzel odds ratio 0.59, 95% confidence interval 0.45 to 0.79) with no evidence for heterogeneity between trials. No unscheduled intravenous fluid infusion therapy was required in any participant in three trials. Eleven trials reported stool output, and data suggested less stool output in the reduced osmolarity ORS group. Vomiting was less frequent in the reduced osmolarity group in the six trials reporting this. Six trials sought hyponatraemia, with events in three studies, but no obvious difference between the two arms. REVIEWER'S CONCLUSIONS: In children admitted to hospital with diarrhoea, reduced osmolarity ORS when compared to WHO standard ORS is associated with fewer unscheduled intravenous fluid infusions, lower stool volume post randomization, and less vomiting. No additional risk of developing hyponatraemia when compared with WHO standard ORS was detected. PMID- 11869640 TI - Treatments for secondary postpartum haemorrhage. AB - BACKGROUND: Secondary postpartum haemorrhage is any abnormal or excessive bleeding from the birth canal occurring between 24 hours and 12 weeks postnatally. In developed countries, two per cent of postnatal women are admitted to hospital with this condition, half of them undergoing uterine surgical evacuation; in developing countries it is a major contributor to maternal death. OBJECTIVES: To evaluate the relative effectiveness and safety of the treatments used for secondary postpartum haemorrhage. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register (May 2001), the Cochrane Controlled Trials Register (The Cochrane Library, issue 2, 2001), MEDLINE (back to 1966) and EMBASE (back to 1988). The National Research Register was also searched. The reference lists of trial reports and reviews were searched by hand. Where possible, further sources were sought from the first named authors of the papers identified. SELECTION CRITERIA: All randomised or quasi randomised comparisons between drug therapies, surgical therapies and placebo or no treatment for the management of secondary postpartum haemorrhage occurring between 24 hours and three months following a pregnancy of at least 24 weeks gestation. DATA COLLECTION AND ANALYSIS: Reports of possibly eligible studies were scrutinised by two investigators. The third investigator acted as an advisor/arbitrator. MAIN RESULTS: Of the 45 papers identified, none met the inclusion criteria. REVIEWER'S CONCLUSIONS: No information is available from randomised controlled trials to inform the management of women with secondary postpartum haemorrhage. This topic may have received little attention because it is perceived as being associated with maternal morbidity rather than mortality in developed countries; it is only recently that the extent and importance of postnatal maternal morbidity has been recognised. A well designed randomised controlled trial comparing the various drug therapies for women with secondary postpartum haemorrhage against each other and against placebo or no treatment groups is needed. PMID- 11869641 TI - Vagus nerve stimulation for partial seizures. AB - BACKGROUND: Vagus nerve stimulation (VNS) has recently been introduced as an adjunct for treating patients with seizures. The aim of this systematic review was to overview the current evidence for the effects of vagus nerve stimulation, when used as an adjunctive treatment for patients with drug-resistant partial epilepsy. OBJECTIVES: To determine the effects of VNS high-level stimulation compared to low-level (presumed subtherapeutic dose) stimulation. SEARCH STRATEGY: We searched the Cochrane Epilepsy Group trials register, MEDLINE (January 1966 to October 2000) and The Cochrane Controlled Trials Register (Cochrane Library Issue 4, 2000). SELECTION CRITERIA: Randomized, double-blind controlled trials of VNS comparing high and low stimulation paradigms. Studies in adults or children with drug-resistant partial seizures. DATA COLLECTION AND ANALYSIS: Two reviewers independently selected trials for inclusion and extracted data. The following outcomes were assessed: (a) 50% or greater reduction in total seizure frequency; (b) treatment withdrawal (any reason); (c) side effects. Primary analyses were intention to treat. Sensitivity best and worst case analyses were also undertaken. Summary odds ratios (ORs) were estimated for each outcome. MAIN RESULTS: Results of the overall efficacy analysis show that VNS stimulation using the high stimulation paradigm was significantly better than low stimulation. The overall OR (95% Confidence Interval (CI)) for 50% responders across all studies is 1.93 (1.1,3.3). This effect did not vary substantially and remained statistically significant for both the best and worst case scenarios. Results for the outcome "withdrawal of allocated treatment" suggest that VNS is well tolerated as no significant difference was found between the high and low stimulation groups, and withdrawals were rare. Statistically significant adverse effects associated with implantation (low versus baseline) were hoarseness, cough, pain and paresthesia. Statistically significant adverse effects associated with stimulation (high versus low) were hoarseness and dyspnea, suggesting the implantation is associated with hoarseness, but the stimulation produces additional hoarseness. REVIEWER'S CONCLUSIONS: VNS for partial seizures appears to be an effective and well tolerated treatment. Adverse effects of hoarseness, cough, pain, paresthesias and dyspnea are associated with the treatment but appear to be reasonably well tolerated as dropouts were rare. Typical central nervous system adverse effects of antiepileptic drugs such as ataxia, dizziness, fatigue, nausea and somnolence were not statistically significantly associated with VNS treatment. PMID- 11869642 TI - Electrical cardioversion for atrial fibrillation and flutter. AB - BACKGROUND: Atrial fibrillation increases the risk of stroke, increases the risk of cognitive impairment, and adversely affects cardiovascular haemodynamics. Electrical cardioversion for atrial fibrillation has been in use since the 1960s; the rationale is that restoration of sinus rhythm improves cardiovascular haemodynamics, reduces the risk of stroke, and obviates the need for long-term anticoagulation. OBJECTIVES: To assess the effects of electrical cardioversion of atrial fibrillation or atrial flutter on the annual risk of thromboembolic events, strokes and mortality (primary outcomes measures), the rate of cognitive decline, quality of life, the use of anticoagulants and the risk of re hospitalisation (secondary outcome measures) in adults (>18 years) with acute, paroxysmal or sustained atrial fibrillation or atrial flutter, of any duration and any aetiology. SEARCH STRATEGY: One reviewer searched the Cochrane Controlled Clinical Trials Register (2000 Issue 4), MEDLINE (1966 to December 2000), EMBASE (1980 to December 2000), CINAHL (1982 to November 2000) and proceedings of the American College of Cardiology (published in the Journal of the American College of Cardiology 1983 to 2000). Reference lists of articles were searched. Personal contact was made with experts in the field. A second reviewer handsearched proceedings of the British Cardiac Society (published in British Heart Journal (1980 to 1995) and in Heart (1995 to May 2001); proceedings of the European Congress of Cardiology and meetings of the Joint Working Groups of the European Society of Cardiology (published in European Heart Journal 1983-2000); scientific sessions of the American Heart Association (published in Circulation 1990-2000). SELECTION CRITERIA: Randomised controlled trial or controlled clinical trials of electrical cardioversion plus 'usual care' versus 'usual care' only, where 'usual care' included any combination of the following: anticoagulants, antiplatelet drugs and drugs for 'rate control', in adults (>18 years) with acute, paroxysmal or sustained atrial fibrillation or atrial flutter, of any duration and any aetiology. DATA COLLECTION AND ANALYSIS: It was planned to extract study data onto data extraction forms. The planned analysis was by the statistical package in RevMan. MAIN RESULTS: No completed randomised trials or controlled clinical trials of electrical cardioversion were found. Two ongoing trials were identified. REVIEWER'S CONCLUSIONS: There were no data from completed randomised controlled trials or controlled clinical trials to either support or refute the use of electrical cardioversion for atrial fibrillation. Randomised trials of electrical cardioversion are required. PMID- 11869643 TI - Enhancing partner support to improve smoking cessation. AB - BACKGROUND: While many cessation programs are available to assist smokers in quitting, research suggests that partner involvement may encourage long-term abstinence. OBJECTIVES: The purpose of this review was to determine if an intervention to enhance partner support helps smoking cessation when added as an adjunct to a smoking cessation program. SEARCH STRATEGY: The search was performed in: Cochrane Tobacco Addiction Group specialized register (Aug 2001), Cochrane controlled trials register (Apr 2000), CDC and Prevention-Tobacco Information and Prevention Database (Jul 2000), MEDLINE (1966-Jul 2000), Cancer Lit (1966-Jul 2000), EMBASE (1974-Apr 2000), CINAHL (1966-Jul 2000), PsycInfo (1861-Oct 2000), ERIC, PsycLit, & Dissertation Abstracts (1861-Dec 1999), SSCI (1972-Jul 2000) and HealthStar (1975-Jul 2000). The search terms used were smoking (prevention, control, therapy), smoking cessation, and support (family, marriage, spouse, partner, sexual partner, buddy, friend, co-habitees, and co-worker). The search was also limited to English language. SELECTION CRITERIA: Randomized controlled trials of smoking cessation interventions that compared an intervention that included a partner support component with an otherwise identical intervention and reported follow-up of 6 months or greater. DATA COLLECTION AND ANALYSIS: Two reviewers independently extracted data using a structured form. A third reviewer was consulted to aid in the resolution of discrepancies. Abstinence and biochemical assessment were the primary outcome measures and were analyzed at two post-treatment intervals: 6-9 months and >12 months. The scores of PIQ (partner interaction questionnaire) were also analyzed to assess partner support. A fixed effects model was used to assess the summary effect of the studies. MAIN RESULTS: A total of 31 articles were identified for this review. Only eight articles (nine studies) met the inclusion criteria. The definition of partner varied among the studies. All studies included data on self reported smoking cessation rates, but there was limited biochemical validation of abstinence rates. The odds ratio for self-reported abstinence at 6-9 months was 1.08 (CI 95%, 0.81 -1.44); and at 12 months post-treatment was 1.0 (CI 95%, 0.75 - 1.34). Of the six studies that measured partner support at follow-up, only two studies reported significant increase in partner support in the intervention groups. REVIEWER'S CONCLUSIONS: We conclude that interventions designed to enhance partner support for smokers in cessation programs did not increase quit rates. Limited data from several of the RCTs suggest that these interventions did not increase partner support either. No conclusions can be made about the impact of partner support on smoking cessation. More systematic intervention to affect partnership significantly should be delivered if partner support were part of an existing cessation program. PMID- 11869644 TI - Disposable surgical face masks for preventing surgical wound infection in clean surgery. AB - BACKGROUND: Surgical face masks were originally developed to contain and filter droplets of microorganisms expelled from the mouth and nasopharynx of healthcare workers during surgery, thereby providing protection for the patient. However there are several ways in which surgical face masks could potentially contribute to contamination of the surgical wound. OBJECTIVES: To identify and review all randomised controlled trials evaluating disposable surgical face masks worn by the surgical team during clean surgery to prevent post-operative surgical wound infection. SEARCH STRATEGY: All relevant publications about disposable surgical face masks were sought through the Specialised Trials Register of the Cochrane Wounds Group (March 2001). Manufacturers and distributors of disposable surgical masks as well as professional organisations including the National Association of Theatre Nurses and the American Operating Room Nurses Association were contacted for details of unpublished and ongoing studies. SELECTION CRITERIA: Randomised controlled trials (RCTs) and quasi-randomised controlled trials comparing the use of disposable surgical masks with the use of no mask were included. DATA COLLECTION AND ANALYSIS: Data were extracted independently by AL and PE. MAIN RESULTS: Two randomised controlled trials were included involving a total of 1453 patients. In a small trial there was a trend towards masks being associated with fewer infections, whereas in a large trial there was no difference in infection rates between the masked and unmasked group. Neither trial accounted for cluster randomisation in the analysis. REVIEWER'S CONCLUSIONS: From the limited results it is unclear whether wearing surgical face masks results in any harm or benefit to the patient undergoing clean surgery. PMID- 11869645 TI - Rapid correction of early metabolic acidaemia in comparison with placebo, no intervention or slow correction in LBW infants. AB - BACKGROUND: Metabolic or mixed (metabolic and respiratory) acidosis are commonly encountered problems in the low birth weight (LBW) infant after delivery, and they may contribute to mortality and morbidity. Causes for the lactic acidosis are multiple and include maternal, placental and fetal factors. It is unclear whether metabolic acidaemia in the first 24 hours of life in LBW infants should be corrected by rapid infusion of alkali. OBJECTIVES: The main objective was to assess the short and long-term effects of the rapid correction of early (first 24 hours) metabolic acidaemia in LBW (<2500g birth weight) neonates. SEARCH STRATEGY: Searches were undertaken of MEDLINE from October 2001 back to 1966 and the Cochrane Controlled Trials Register (Cochrane Library, Issue 4, 2001). The title and abstract of each retrieved study were examined to assess eligibility. If there was uncertainty, the full paper was examined. SELECTION CRITERIA: Types of studies All randomised controlled trials where short or long term effects of treatment with alkalising agents by rapid infusion were compared with placebo or no treatment, or where rapid infusion of alkalising agents was compared with slow infusion. Types of participants Newborn infants with birth weight <2500g and less than 24 hours of age with proven metabolic acidaemia (on arterial blood gas). Types of interventions Rapid correction of acidaemia with alkalising agents (sodium bicarbonate and/or THAM) given as a bolus over 5 minutes or less compared with either placebo, no intervention or slow infusion (>5 minutes). Types of outcome measures 1) maximal oxygen requirement in first 24 hours 2) duration of oxygen therapy 3) need for and duration of assisted ventilation 4) intraventricular haemorrhage and/or periventricular leucomalacia 5) survival to discharge 6) long term survival (to 24 months of age) 7) neurological and developmental outcome at 24 months of age DATA COLLECTION AND ANALYSIS: Each reviewer assessed eligibility, trial quality and extracted data separately, then compared and resolved differences. Study authors were contacted for additional information if necessary. MAIN RESULTS: No studies were found meeting the criteria for inclusion in this review. REVIEWER'S CONCLUSIONS: There is no evidence available from randomised controlled trials to support or refute the rapid correction of metabolic acidaemia, in LBW infants in the first 24 hours of life, as compared with slow or no correction. PMID- 11869646 TI - Cardioselective beta-blockers for reversible airway disease. AB - BACKGROUND: Beta-blocker therapy has mortality benefit in patients with hypertension, heart failure and coronary artery disease, as well as during the perioperative period. These drugs have traditionally been considered contraindicated in patients with reversible airway disease. OBJECTIVES: To assess the effect of cardioselective beta1-blockers on respiratory function of patients with reversible airway disease. Reversible airway disease was defined as asthma or chronic obstructive pulmonary disease with a reversible obstructive component. SEARCH STRATEGY: A comprehensive search of EMBASE, MEDLINE and CINAHL was performed using the Cochrane Airways Group registry to identify randomized blinded placebo-controlled trials from 1966 to May 2001. The search was completed using the terms: asthma*, bronchial hyperreactivity*, respiratory sounds*, wheez*, obstructive lung disease* or obstructive airway disease*, and adrenergic antagonist*, sympatholytic* or adrenergic receptor block*. We did not exclude trials on the basis of language. SELECTION CRITERIA: Randomized, blinded, placebo controlled trials of single dose or longer duration that studied the effects of cardioselective beta1-blockers on the forced expiratory volume in 1 second (FEV1), symptoms and use of short-acting inhaled beta2-agonists, in patients with reversible airway disease. Reversible airway disease was documented by response to methacholine challenge, by an increase in FEV1 of at least 15% to beta2 agonist administration, or the presence of asthma as defined by the American Thoracic Society. DATA COLLECTION AND ANALYSIS: Two independent reviewers extracted data from the selected articles, reconciling differences by consensus. Cardioselective beta1-blockers were divided into 2 groups, those with or without intrinsic sympathomimetic activity (ISA). Two interventions studied were the administration of beta1-blocker, given either as a single dose or for longer duration, and the use of beta2-agonist given after the study drug. MAIN RESULTS: Nineteen studies for single-dose treatment and 10 for treatment of longer duration met selection criteria. The patients had mild-moderate airways obstruction. For cardioselective beta1-blockers taken as a group, administration of a single dose was associated with a 7.98% (CI, 6.19 to 9.77%) reduction in FEV1, but with a 13.16% (CI, 10.76 to 15.56%) increase in beta2-agonist response, as compared to placebo. There was no increase in symptoms. After treatment lasting a few days to a few weeks, there was no decrement in FEV1 compared to placebo and no increase in symptoms or inhaler use. Regular use of cardioselective beta1-blockers without ISA produced a 13.13% (CI, 5.97 to 20.30) increase in beta2-agonist response compared to placebo, a response not seen with beta1-blockers containing ISA (-0.60% [CI, -11.7 to +10.5%]). REVIEWER'S CONCLUSIONS: Cardioselective beta1-blockers, given to patients with mild-moderate reversible airway disease, do not produce clinically significant adverse respiratory effects in the short term. It is not possible to comment on their effects in patient with more severe or less reversible disease, or on their effect on the frequency or severity of acute exacerbations. Given their demonstrated benefit in conditions such as heart failure, coronary artery disease and hypertension, cardioselective beta1-blockers should not be withheld from patients with mild-moderate reversible airway disease. PMID- 11869647 TI - Inhaled hyperosmolar agents for bronchiectasis. AB - BACKGROUND: Mucus retention in the lungs is a prominent feature of bronchiectasis. The stagnant mucus becomes chronically colonised with bacteria, which elicit a host neutrophilic response. This fails to eliminate the bacteria, and the large concentration of host-derived protease may contribute to the airway damage. The sensation of retained mucus is itself a cause of suffering, and the failure to maintain airway sterility probably contributes to the frequent respiratory infections experienced by many patients. Hypertonic saline inhalation is known to accelerate tracheobronchial clearance in many conditions, probably by inducing a liquid flux into the airway surface, which alters mucus rheology in a way favourable to mucociliary clearance. Inhaled dry powder mannitol has a similar effect. Such agents are an attractive approach to the problem of mucostasis, and deserve further clinical evaluation. OBJECTIVES: To determine whether inhaled hyperosmolar substances are efficacious in the treatment of bronchiectasis SEARCH STRATEGY: MEDLINE and Cochrane databases were searched, and leaders in the field contacted. SELECTION CRITERIA: Any trial using hyperosmolar inhalation in patients with bronchiectasis not caused by cystic fibrosis. DATA COLLECTION AND ANALYSIS: One reference were identified by the searches conducted. MAIN RESULTS: Only one trial was identified, a crossover study of 11 patients with bronchiectasis. The outcome measure was tracheobronchial clearance of a particulate radioaerosol after inhalation of dry mannitol on a single occasion, with appropriate controls. Airway clearance doubled in the central and intermediate regions of the lung, but not in the peripheral region, after mannitol administration. No side effects were observed, but two patients were premedicated with nedocromil to prevent bronchospasm. A further search conducted in September 2001 did not identify any further studies. REVIEWER'S CONCLUSIONS: Dry powder mannitol has been shown to improve tracheobronchial clearance in bronchiectasis, as well as cystic fibrosis, asthmatics, and normal subjects. It is not yet available for clinical use. Hypertonic saline has not been specifically tested in bronchiectasis, but improve clearance in these other conditions and in chronic bronchitis. Longer term randomised controlled studies of mannitol and hypertonic saline with clinical endpoints are now needed. PMID- 11869648 TI - Macrolides for chronic asthma. AB - BACKGROUND: Asthma is a chronic disease of the airways in which inflammation of the respiratory mucosa plays a crucial role. The mechanisms responsible for the maintaining of this inflammatory response are only partially known and there is evidence that a role could be paid by chronic infection by intracellular pathogens (such as Chlamydia pneumoniae). Macrolides are antibiotics with both antimicrobial and anti-inflammatory activities and thus their use in asthmatic patients could lead to reduction of the airways inflammation and therefore improvement of symptoms and pulmonary function. OBJECTIVES: To determine whether macrolides are effective in the management of patients with chronic asthma. SEARCH STRATEGY: We searched MEDLINE, EMBASE and CINAHL up to May 2001. This was also supplemented by manually searching bibliographies of previously published reviews, conference proceedings, and contacting study authors. All languages were included in the initial search. SELECTION CRITERIA: Randomised, controlled clinical trials involving both children and adult patients with chronic asthma treated with macrolides for more than 4 weeks, versus placebo. DATA COLLECTION AND ANALYSIS: Two reviewers independently examined all identified articles. Two reviewers reviewed the full text of any potentially relevant article independently. MAIN RESULTS: The initial search retrieved 95 studies. Preliminary evaluation identified 20 studies that were potentially eligible. Five (357 patients) met the entry criteria. The entry criteria for the primary trials differed, but all recruited a specific subgroup of patients (eg severe oral steroid dependent, aspirin intolerant or evidence of Chlamydia pnuemoniae infection). There was a positive effect on symptoms (Standardised Mean Difference -1.25, 95% Confidence Intervals (CI) -1.80, -0.70) and markers of eosinophilic inflammation; eg sputum eosinophils Weighted Mean difference -78.5, 95%CI -90.8, 66.1). Tests of oral corticosteroid-sparing effects have not yet been performed on the newer agents such as roxithromycin and clarithromycin. REVIEWER'S CONCLUSIONS: Considering the small number of patients studied, there is insufficient evidence to support or to refute the use of macrolides in patients with chronic asthma. Further studies are needed in particular to clarify the potential role of macrolides in some subgroups of asthmatics such as those with evidence of chronic bacterial infection. PMID- 11869649 TI - Perioperative fluid volume optimization following proximal femoral fracture. AB - BACKGROUND: Proximal Femoral Fracture (PFF) or 'hip fracture' is a frequent injury, and adverse outcomes are common. Many patients are elderly, with significant comorbidity. Several factors suggest the importance of developing appropriate techniques to optimize intravascular fluid volume. These may include protocols that enhance the efficacy of clinicians' assessments, invasive techniques such as oesophageal Doppler or central venous pressure monitoring, or advanced non-invasive techniques such as plethysmographic pulse volume determination. OBJECTIVES: To determine the optimal method of fluid volume optimization for adult patients undergoing surgical repair of hip fracture. Comparisons of fluid types (e.g. crystalloid vs. colloid) or of blood transfusion strategies or of other pharmacological interventions (e.g. inotropes) are not considered in this review. SEARCH STRATEGY: Randomized controlled trials (RCTs) since 1985 were identified by searching MEDLINE, EMBASE, the Cochrane Library, the Cochrane Anaesthesia Group's specialized Controlled Trials Register and bibliographies of retrieved articles. Relevant journals and conference proceedings were handsearched. SELECTION CRITERIA: RCTs comparing a fluid optimization intervention with normal practice (control) or with another fluid optimization intervention, in patients following PFF undergoing surgery of any type under anaesthesia of any type. DATA COLLECTION AND ANALYSIS: Searches and exclusion of clearly irrelevant articles were performed by one reviewer. Two reviewers examined independently the remaining studies, extracting study quality and results data. A wide range of short- and long-term outcome data was sought. Study quality was assessed using a ten-point instrument and studies were excluded if they did not meet the study criteria or if results were likely to be biased. Due to a lack of consistency in reporting, combination of data was not generally possible. MAIN RESULTS: Searches identified only four trials, of which two studies, randomizing a total of 130 patients, were of adequate quality and addressed the review question. Both studies were of invasive advanced haemodynamic monitoring during the intraoperative period only. One study randomized patients to 'normal care' or optimization using oesophageal Doppler; the second study randomized patients to 'normal care', oesophageal Doppler or central venous pressure monitoring. In each study, invasive monitoring led to a significant increase in volume of fluid infused and a reduction in length of hospital stay. The pooled Peto odds ratio for in-hospital fatality was 1.44 (95% confidence interval 0.45-4.62). Neither study followed patients beyond hospital discharge or assessed functional outcomes (for example return to previous accommodation). No serious complications were reported to be directly attributable to the monitoring interventions. There were no studies of protocol guided fluid optimization or of advanced non-invasive techniques. REVIEWER'S CONCLUSIONS: Invasive methods of fluid optimization during surgery may shorten hospital stay, but their effects on other important, patient-centred, longer-term outcomes are uncertain. An adverse effect on fatality cannot be excluded. Other fluid optimization techniques have not been evaluated. The lack of randomized studies of adequate quality addressing this important question is disappointing given the high incidence and frequently adverse outcome of PFF. More research is needed. PMID- 11869650 TI - Pharmacological interventions for recurrent abdominal pain (RAP) in childhood. AB - BACKGROUND: Between 4% and 25% of school age children complain of recurrent abdominal pain (RAP) of sufficient severity to interfere with daily activities. For the majority of such children no organic cause for their pain can be found on physical examination or investigation. Although most children are likely managed by reassurance and simple measures, a large range of interventions has been recommended. OBJECTIVES: To determine the effectiveness of medication for recurrent abdominal pain in school-age children. SEARCH STRATEGY: The Cochrane Library (CENTRAL), MEDLINE, EMBASE, CINAHL, ERIC, PsycLIT, LILACS and JICST were searched using a strategy combining (Recurrent OR synonyms) AND (Abdomen OR synonyms) AND (Pain OR synonyms). Where appropriate search filters were employed. In addition, researchers working in this area were asked to identify relevant studies. SELECTION CRITERIA: Any study in which the majority of participants were school age children fulfilling standard criteria for RAP, and who were allocated by random or quasi-random methods to any drug treatment compared with a placebo or no treatment. DATA COLLECTION AND ANALYSIS: References identified by the searches were screened against the inclusion criteria by two independent reviewers. MAIN RESULTS: Only one trial met the inclusion criteria. This cross over trial in 14 children who met suggested criteria for "abdominal migraine" compared pizotifen and placebo, each given for one month with no washout period. Participants reported a mean of 8.21 (95% CI 2.93, 13.48) fewer days of pain while taking the active drug. They also reported that the mean difference on an "Index of Severity" was -16.21 (95% CI -26.51, -5.90) and on an "Index of Misery" was -56.07 (95% CI -94.07, -18.07). REVIEWER'S CONCLUSIONS: There is little evidence to suggest that recommended drugs are effective in the management of RAP. At present there seems little justification for the use of these drugs other than in clinical trials. There is an urgent need for trials of all suggested pharmacologic interventions in children with RAP. PMID- 11869651 TI - Furosemide for transient tachypnea of the newborn. AB - BACKGROUND: Transient tachypnea of the newborn results from delayed clearance of lung liquid and is a common cause of admission of full term infants to neonatal intensive care units. The condition is particularly common after elective Caesarean section. Conventional treatment involves appropriate oxygen administration and continuous positive airway pressure in some cases. Most infants receive antibiotic therapy. Hastening the clearance of lung liquid should shorten the duration of the symptoms and reduce complications. OBJECTIVES: To determine whether furosemide reduces the duration of oxygen therapy and respiratory symptoms and shortens hospital stay in term infants with transient tachypnea of the newborn. SEARCH STRATEGY: We searched the Cochrane Controlled Trials Register, PubMed and EMBASE. The primary author and experts in the field were contacted. SELECTION CRITERIA: Randomised or quasi-randomised controlled trials. Infants of less than 7 days of age, born after 37 or more weeks of gestation with the clinical picture of transient tachypnea of the newborn. Intravenous, oral or nebulized furosemide compared to placebo or no diuretic in the first 7 days. DATA COLLECTION AND ANALYSIS: Two reviewers assessed trial quality in each potentially eligible manuscript and two reviewers extracted data. MAIN RESULTS: Searching revealed only one randomised trial which was methodologically sound. This recruited 50 infants with transient tachypnea. Infants were randomised to receive oral furosemide 2 mg/kg followed by 1 mg/kg 12 hours later, or placebo. Weight loss in the first 24 hours was greater in the furosemide treated group but there was no evidence of a difference between the groups in duration of tachypnea or severity of symptoms or length of hospitalization. The study was methodologically satisfactory. REVIEWER'S CONCLUSIONS: Oral furosemide cannot be recommended as treatment for transient tachypnea of the newborn and it should not be used unless additional data become available. The question remains as to whether intravenous furosemide given to the infant (or even to the mother before Caesarean section) might shorten the duration of the illness. As elective Caesarean section continues at a high level, these two interventions might be worthy of trials. PMID- 11869652 TI - Benperidol for schizophrenia. AB - BACKGROUND: Benperidol is a relatively old antipsychotic drug, marketed since 1966, used in Germany for 30 years, but also available in Belgium, Greece, Italy, the Netherlands and the UK. Benperidol is a butyrophenone antipsychotic, with the highest neuroleptic potency in terms of D2 receptor blockade. Those taking it are, therefore, reputed to be at high risk of extrapyramidal side effects, but benperidol's unusual profile may render it of value to subgroups of people with schizophrenia. OBJECTIVES: To examine the clinical effects and safety of benperidol for those with schizophrenia and schizophrenia-like psychoses. SEARCH STRATEGY: The reviewers searched the Cochrane Schizophrenia Group's register (January 2001) which includes relevant randomised controlled trials from the bibliographic databases Biological Abstracts, CINAHL, The Cochrane Library, EMBASE, MEDLINE, PsycLIT, LILACS, PSYNDEX, Sociological Abstracts and Sociofile. We also searched the references of all included studies and contacted pharmaceutical companies and authors of included studies in order to identify further trials. SELECTION CRITERIA: Randomised controlled trials that compared benperidol with other treatments for people with schizophrenia and/or schizophrenia-like psychoses. DATA COLLECTION AND ANALYSIS: Citations and, where possible, abstracts were independently inspected by two reviewers and papers were ordered, re-inspected and quality assessed. We independently extracted data but excluded data if loss to follow up was greater than 50%. For homogeneous dichotomous data, we calculated the relative risk (RR), the 95% confidence interval (CI) and, where appropriate, the number needed to treat/harm (NNT/H), on an intention-to-treat basis. MAIN RESULTS: We identified only one unpublished poorly randomised controlled trial (N=40, duration 30 days, comparison perphenazine). Although benperidol was inferior to perphenazine (1 RCT, N=40, global state no better or worse RR 8.0 CI 2.1 to 30, NNH 1.4 CI 1 to 2) poor reporting suggests that an overestimate of effect is likely. It was not possible to report other outcomes. REVIEWER'S CONCLUSIONS: Currently, there are insufficient data from randomised trials to assess the clinical effects of benperidol. This interesting compound merits further research. PMID- 11869653 TI - Addition of anti-leukotriene agents to inhaled corticosteroids for chronic asthma. AB - BACKGROUND: Anti-leukotriene (AL) agents are being considered as "add-on" therapy to inhaled corticosteroids (ICS), in chronic asthma. OBJECTIVES: To examine the safety and efficacy of daily AL plus ICS compared to ICS alone, and determine the corticosteroid-sparing effect of AL when added to ICS in chronic asthma. SEARCH STRATEGY: We searched Medline, Embase, Cinahl (until September 2001), reference lists of review articles and trials, contacted international headquarters of AL manufacturers and ATS meeting abstracts (1998-2000). SELECTION CRITERIA: Randomised placebo-controlled trials of asthmatics aged 2 years and older with at least one month intervention. DATA COLLECTION AND ANALYSIS: Two reviewers assessed quality and extracted data independently. Trials were grouped by asthma control at baseline (symptomatic or well-controlled) and dose of ICS in the control group (same or double). MAIN RESULTS: Of 438 citations, 13 (12 adult and 1 paediatric) trials met inclusion criteria. Seven were published in full-text. In symptomatic patients, addition of licensed doses of anti-leukotrienes to ICS resulted in a non-significant reduction in the risk of exacerbations requiring systemic steroids: Relative Risk (RR) 0.61; 95% Confidence Interval (CI) 0.36,1.05). A modest improvement group difference in PEF was seen (Weighted Mean Difference (WMD) 7.71 L/min; 95%CI 2.98, 12.44 L/min) together with beta2-agonist use (WMD= -0.32 puffs/day; 95%CI -0.0.08, -0.56). No trials that compared the use of licensed doses of anti-leukotrienes with doubling-dose of inhaled glucocorticoids could be pooled. In ICS-sparing studies in patients who were well controlled at baseline, addition of anti-leukotrienes produced no overall difference in dose of inhaled glucocorticoids (WMD -44.4 mcg/d, 95%CI -147.9, 59.0 mcg/d), but it was associated with fewer withdrawals due to poor asthma control (RR= 0.56, 95%CI 0.35, 0.89). REVIEWER'S CONCLUSIONS: There is insufficient evidence to firmly support the use of licensed doses of anti leukotrienes as add-on therapy to inhaled glucocorticoids. Addition of anti leukotrienes to inhaled glucocorticoids may slightly improve asthma control, but the available data do not permit this strategy to be recommended as a substitute for increasing the dose of inhaled glucocorticoids. Addition of anti-leukotrienes may be associated with superior asthma control after glucocorticoid tapering, but a glucocorticoid-sparing effect cannot be quantified at present. PMID- 11869654 TI - Blood transfusion for preventing stroke in people with sickle cell disease. AB - BACKGROUND: Sickle cell disease is a common inherited haemoglobin disorder. The abnormal haemoglobin causes distortion of red blood cells, anaemia, vaso occlusion and dysfunction in virtually any organ system in the body. Stroke occurs in around 10% of children with sickle cell anaemia, and recurrences after a first stroke are likely. Chronic blood transfusion regimes are often used in an attempt to dilute the sickled red blood cells, thus reducing the risk of vaso occlusion and stroke. However, the side-effects of such regimens can be severe. OBJECTIVES: To assess the relative risks and benefits of chronic blood transfusion regimes in patients with sickle cell disease to prevent a first stroke or further strokes. SEARCH STRATEGY: We searched the Cochrane Cystic Fibrosis and Genetic Disorders Group specialist register which comprises references identified from comprehensive electronic database searches, handsearching relevant journals and handsearching abstract books of conference proceedings. Date of the most recent search of the Group's specialised register: July 2001 SELECTION CRITERIA: All those randomised or quasi-randomised controlled trials in which blood transfusion as a preventative measure for stroke in patients with sickle cell disease are compared to an alternative treatment or to no treatment. DATA COLLECTION AND ANALYSIS: Both reviewers independently assessed trial quality and extracted data from the study included. MAIN RESULTS: One trial was identified by the initial search and this met the inclusion criteria for the review. The trial compared a chronic transfusion regime to maintain sickle haemoglobin at less than 30% with standard care in 130 children with sickle cell disease who were judged to be at high risk of a first stroke through transcranial doppler ultrasonography. Eleven children in the standard care group suffered a stroke during the trial, compared to only one in the transfusion group. Because of this 92% relative risk reduction, the trial was terminated 16 months early. It had been planned that all patients would be treated for 30 months, but median follow-up only 21.1 months. However, a high rate of complications such as iron overload, alloimmunisation and transfusion reactions were seen in the children who were receiving transfusions. No randomised controlled trials were identified which investigated use of transfusion for preventing recurrence of stroke. REVIEWER'S CONCLUSIONS: While the included study demonstrated a significantly reduced risk of stroke in patients receiving regular blood transfusions, the degree of risk must be balanced against the burden of a chronic transfusion regime. Further research is required to establish the use of transfusion in preventing secondary stroke, the age, or length of time after an event, at which transfusion can safely be stopped, and to further define risk factors for stroke in order to reduce the chance of unnecessarily putting children onto a chronic transfusion regime. PMID- 11869655 TI - The Epley (canalith repositioning) manoeuvre for benign paroxysmal positional vertigo. AB - BACKGROUND: Benign paroxysmal positional vertigo (BPPV) is a syndrome characterised by short-lived episodes of vertigo in association with rapid changes in head position. It is a common cause of vertigo presenting to primary care and specialist otolaryngology clinics. Current treatment approaches include rehabilitative exercises and physical manoeuvres including the Epley manoeuvre. OBJECTIVES: To assess the effectiveness of the Epley manoeuvre compared to other treatments available for posterior canal benign paroxysmal positional vertigo, or no treatment. SEARCH STRATEGY: The Cochrane Controlled Trials Register (Cochrane Library, Issue 2 2001), MEDLINE (1966 onwards), EMBASE (1974 onwards), and reference lists of identified publications. Date of the most recent search was June 2001. SELECTION CRITERIA: Randomised trials of adults diagnosed with posterior canal BPPV (including a positive Dix-Hallpike test). Comparisons sought: - Epley manoeuvre versus placebo. - Epley manoeuvre versus untreated controls. Epley manoeuvre versus other active treatment. Outcome measures that were considered include: frequency and severity of attacks of vertigo; proportion of patients improved by each intervention; and conversion of a "positive" Dix Hallpike test to a "negative" Dix-Hallpike test DATA COLLECTION AND ANALYSIS: Both reviewers independently extracted data and assessed trials for quality. MAIN RESULTS: Eleven trials were identified but nine studies were excluded because of a high risk of bias, leaving two trials in the review. Trials were mainly excluded because of inadequate concealment during randomisation, or failure to blind outcome assessors. Both studies included in the review (Lynn 1995, Froehling 2000) addressed the efficacy of the Epley manoeuvre against a sham manoeuvre by comparing the proportion of subjects in each group who had complete resolution of their symptoms, and who converted from a positive to negative Dix Hallpike test. Individual and pooled data showed a statistically significant effect in favour of the Epley manoeuvre over controls. There were no serious adverse effects of treatment. REVIEWER'S CONCLUSIONS: ~bullet~There is some evidence that the Epley manoeuvre is a safe effective treatment for posterior canal BPPV, although this is based on the results of only two small randomised controlled trials with relatively short follow up. ~bullet~There is no good evidence that the Epley manoeuvre provides a long term resolution of symptoms. ~bullet~There is no good evidence comparing the Epley manoeuvre with other physical, medical or surgical therapy for posterior canal BPPV. PMID- 11869656 TI - Tryptophan and 5-hydroxytryptophan for depression. AB - BACKGROUND: 5 Hydroxytryptophan (5-HTP) and tryptophan are so-called natural alternatives to traditional antidepressants, used to treat unipolar depression and dysthymia. OBJECTIVES: To determine whether 5-HTP and tryptophan are more effective than placebo, and whether they are safe to use to treat depressive disorders in adults. SEARCH STRATEGY: Trials were searched in computerized general (Medline, Psychlit, and Embase) and specialized databases (Cochrane Controlled Clinical Trials Register, Cochrane Collaboration Depression, Anxiety and Neurosis Controlled Trial Register); by checking reference lists of relevant articles; by handsearching relevant specialist journals; and by contacting relevant authors where appropriate. Publications in all languages were sought. SELECTION CRITERIA: Trials were included if they were randomized, included patients with unipolar depression or dysthymia, compared preparations of 5-HTP or tryptophan with placebo, and included clinical outcomes assessed by scales assessing depressive symptoms. DATA COLLECTION AND ANALYSIS: Data was extracted independently by the three reviewers, onto data collection forms. Inclusion criteria were applied to all potential studies independently and a coefficient of agreement (Kappa) was calculated for them. Disagreement was resolved by reaching consensus. Trial quality was scored according to risk of bias. Analysis for 5-HTP and tryptophan were combined due to the small number of included trials. MAIN RESULTS: 108 trials were located using the specified search strategy. Of these, only two trials, involving a total of 64 patients, were of sufficient quality to meet inclusion criteria. The available evidence suggests these substances were better than placebo at alleviating depression (Peto Odds Ratio 4.10; 95% confidence interval 1.28-13.15; RD 0.36; NNT 2.78). However, the evidence was of insufficient quality to be conclusive. REVIEWER'S CONCLUSIONS: A large number of studies appear to address the research questions, but few are of sufficient quality to be reliable. Available evidence does suggest these substances are better than placebo at alleviating depression. Further studies are needed to evaluate the efficacy and safety of 5-HTP and tryptophan before their widespread use can be recommended. The possible association between these substances and the potentially fatal Eosinophilia-Myalgia Syndrome has not been elucidated. Because alternative antidepressants exist which have been proven to be effective and safe the clinical usefulness of 5-HTP and tryptophan is limited at present. PMID- 11869657 TI - Horse chestnut seed extract for chronic venous insufficiency. AB - BACKGROUND: Conservative therapy of chronic venous insufficiency (CVI) consists largely of compression treatment. However, this often causes discomfort and has been associated with poor compliance, which renders oral drug treatment an attractive option. OBJECTIVES: To review the evidence from rigorous clinical trials assessing the efficacy and safety of oral horse chestnut seed extract (HCSE) versus placebo, or reference therapy for the treatment of CVI. SEARCH STRATEGY: Publications describing randomised controlled trials (RCTs) of HCSE for chronic venous insufficiency were sought through EMBASE, MEDLINE, Amed, and Phytobase (from inception to January 2001), The Cochrane Controlled Trials Register (Issue 1, 2001) and the Specialised Trials Register of the Cochrane Peripheral Vascular Diseases Group (April 2001). Manufacturers of HCSE preparations and experts on the subject were contacted and asked to contribute published and unpublished material. There were no restrictions on the language of publication. SELECTION CRITERIA: Randomised controlled trials of oral HCSE mono preparations for patients with CVI were included. Trials comparing HCSE with placebo or reference medications were included. Trials assessing HCSE as one of several active components in a combination preparation or as a part of a combination treatment were excluded. DATA COLLECTION AND ANALYSIS: Data were extracted systematically and methodological quality was evaluated using a standard scoring system. The screening of studies, selection, data extraction and the assessment of methodological quality were performed independently by two reviewers. Disagreements concerning evaluation of individual trials were resolved through discussion. MAIN RESULTS: Overall, the included placebo controlled trials suggest an improvement in CVI related signs and symptoms. Leg pain was assessed in six placebo-controlled trials that reported a significant reduction of leg pain in the HCSE groups compared with the placebo groups. One trial, which reported adequate data suggested a weighted mean difference (WMD) of 42.4 mm [95% confidence interval (CI) 34.9-49.9] measured on a 100 mm visual analogue scale. Leg volume was assessed in five placebo-controlled trials. Meta-analysis of four trials (n = 239) reporting adequate data suggested a significant reduction in favour of HCSE compared with placebo (WMD 58.6 ml [95% CI 24.9-92.2]). One trial indicated that HCSE may be as effective as treatment with compression stockings. Adverse effects are usually mild and infrequent. REVIEWER'S CONCLUSIONS: The evidence presented implies that HCSE is an efficacious and safe short-term treatment for CVI. However several caveats exist and more rigorous RCTs are required to assess the efficacy of this treatment option. PMID- 11869658 TI - Condom effectiveness in reducing heterosexual HIV transmission. AB - BACKGROUND: The amount of protection that condoms provide for HIV and other sexually transmitted infections is unknown. Cohort studies of sexually active HIV serodiscordant couples with follow-up of the seronegative partner, provide a situation in which a seronegative partner has known exposure to the disease and disease incidence can be estimated. When some individuals use condoms and some do not, namely some individuals use condoms 100% of the time and some never use (0%) condoms, condom effectiveness can be estimated by comparing the two incidence rates. Condom effectiveness is the proportionate reduction in disease due to the use of condoms. OBJECTIVES: The objective of this review is to estimate condom effectiveness in reducing heterosexual transmission of HIV. SEARCH STRATEGY: Studies were located using electronic databases (AIDSLINE, CINAHL, Embase, and MEDLINE) and handsearched reference lists. SELECTION CRITERIA: For inclusion, studies had to have: (1) data concerning sexually active HIV serodiscordant heterosexual couples, (2) a longitudinal study design, (3) HIV status determined by serology, and (4) contain condom usage information on a cohort of always (100%) or never (0%) condom users. DATA COLLECTION AND ANALYSIS: Studies identified through the above search strategy that met the inclusion criteria were reviewed for inclusion in the analysis. Sample sizes, number of seroconversions, and the person-years of disease-free exposure time were recorded for each cohort. If available, the direction of transmission in the cohort (male-to-female, female to-male), date of study enrollment, source of infection in the index case, and the presence of other STDs was recorded. Duplicate reports on the same cohort and studies with incomplete or nonsepecific information were excluded. HIV incidence was estimated from the cohorts of "always" users and for the cohorts of "never" users. Effectiveness was estimated from these two incidence estimates. MAIN RESULTS: Of the 4709 references that were initially identified, 14 were included in the final analysis. There were 13 cohorts of "always" users that yielded an homogeneous HIV incidence estimate of 1.14 [95% C.I.:.56, 2.04] per 100 person years. There were 10 cohorts of "never" users that appeared to be heterogeneous. The studies with the longest follow-up time, consisting mainly of studies of partners of hemophiliac and transfusion patients, yielded an HIV incidence estimate of 5.75 [95% C.I.: 3.16, 9.66] per 100 person-years. Overall effectiveness, the proportionate reduction in HIV seroconversion with condom use, is approximately 80%. REVIEWER'S CONCLUSIONS: This review indicates that consistent use of condoms results in 80% reduction in HIV incidence. Consistent use is defined as using a condom for all acts of penetrative vaginal intercourse. Because the studies used in this review did not report on the "correctness" of use, namely whether condoms were used correctly and perfectly for each and every act of intercourse, effectiveness and not efficacy is estimated. Also, this estimate refers in general to the male condom and not specifically to the latex condom, since studies also tended not to specify the type of condom that was used. Thus, condom effectiveness is similar to, although lower than, that for contraception. PMID- 11869659 TI - Methenamine hippurate for preventing urinary tract infections. AB - BACKGROUND: Methenamine salts are often used for the prevention of urinary tract infection (UTI). OBJECTIVES: To assess the effectiveness of methenamine hippurate in preventing UTI. SEARCH STRATEGY: Published and unpublished randomised controlled trials were identified from the Cochrane Controlled Trials Register, MEDLINE, EMBASE, CINAHL, Current Contents, reference lists of review articles and retrieved trials. The manufacturers' of methenamine salts were contacted for unpublished studies and contact was made with known investigators in the area. SELECTION CRITERIA: Randomised and quasi-randomised trials of methenamine hippurate used for the prevention of UTIs in all population groups were eligible for inclusion. A comparison with a control (no treatment) group was a prerequisite to selection. DATA COLLECTION AND ANALYSIS: Two reviewers (BL and TB) performed independent assessment and data extraction using a standardised format. Discrepancies, methodological and interpretative issues were discussed with JS or JC. An exploration of heterogeneity as well as a detailed description of results grouped by population was conducted. MAIN RESULTS: Eleven studies met the inclusion criteria. All trials were included in a descriptive analysis. Seven trials were included in meta-analyses. Four trials (199 patients) studied symptomatic bacteriuria and six trials (341 patients) studied bacteriuria as an outcome measure. Overall, trial quality was poor. The direction of six of the pooled trials was towards a favourable treatment effect from methenamine hippurate. Interpretation of the pooled estimates was not done in view of underlying heterogeneity. The study by Pettersson 1989 explained some, but not all, of the underlying heterogeneity. This study differed from all others by including patients with known upper renal tract abnormalities. Adverse reactions were mentioned by 10 studies. The rate of adverse events was low. REVIEWER'S CONCLUSIONS: There is not enough evidence to conclusively support the use of methenamine hippurate for urinary prophylaxis. An exploration of heterogeneity raises the (hypothesis generating) possibility that methenamine hippurate may have some efficacy in patients without but not in patients with known upper renal tract abnormality (with asymptomatic bacteriuria as the outcome measure). Due to the small sample size and methodological problems within the studies involved, interpretation of these data should be done cautiously. The rate of adverse events reported by the trials was low, which suggests that current usage is unlikely to be causing significant harm. PMID- 11869660 TI - Methadone at tapered doses for the management of opioid withdrawal. AB - BACKGROUND: Despite use in many countries of tapered methadone for detoxification from opiate, the evidence of efficacy to prevent relapse and promote lifestyle change has not been systematically evaluated. OBJECTIVES: To determine whether tapered methadone is effective to manage withdrawal from opioids. SEARCH STRATEGY: We searched: Cochrane Controlled Trial Register (Issue 1, 2000), MEDLINE (OVID 1966-2000), EMBASE (1980-2000); reference list of relevant articles; personal communication; conference abstracts; trials from pharmaceutical industry; Internet. SELECTION CRITERIA: All randomised controlled trials on the use of tapered methadone versus all other detoxification treatments, placebo and different modalities of methadone detoxification programs for the treatment of opiate withdrawal. DATA COLLECTION AND ANALYSIS: One reviewer assessed studies for inclusion and undertook data extraction. The overall process were confirmed by consultation between reviewers. MAIN RESULTS: 20 studies were included, with 1357 people randomised. 10 studies compared methadone with adrenergic agonists, 7 compared different modalities of methadone detoxification, 2 compared methadone with other opioid agonists, 1 study compared methadone with chlordiazepoxide one with placebo. 10 studies that compared methadone with adrenergic agonists showed clinical differences of the treatments; 6 studies that compared different methadone schedules, showed different withdrawal responses; 2 studies that compared methadone and other opioid agonists showed that methadyl acetate performed similarly to methadone, while propoxyphene produced more severe withdrawal symptoms and more drop-outs than methadone; chlordiazepoxide and methadone (1 study) resulted similar in terms of overall effectiveness; more severe withdrawal and more drop outs in the placebo group than in methadone one (1 study). The results indicate that tapered methadone and other medications used are effective although symptoms experienced differed according to the medication used and the program adopted. The medications are similar in terms of overall effectiveness. Improvements were achieved when other services supporting services were offered contemporaneously with detoxification. REVIEWER'S CONCLUSIONS: Data from literature are hardly comparable; programs vary widely with regard to duration, design and treatment objectives, along with reporting impairing the application of meta-analysis. The studies included confirm that slow tapering with temporary substitution of long acting opioid can reduce withdrawal severity. Nevertheless the majority of patients relapsed to heroin use, abstinence cannot be considered a goal as heroin dependence is a chronic, relapsing disorder and the purpose of detoxification should be to remove or reduce dependence on heroin in a controlled and human fashion and not a treatment for heroin dependence. PMID- 11869661 TI - Hydromorphone for acute and chronic pain. AB - BACKGROUND: While morphine is the gold standard for the management of severe cancer pain, some patients either do not achieve adequate analgesia, or suffer intolerable morphine-related toxicity. For these patients alternatives such as hydromorphone are recommended. However, there appear to be gaps in our understanding of the efficacy and potency of hydromorphone. OBJECTIVES: This review explores and assesses the evidence for the efficacy of hydromorphone in the management of pain. SEARCH STRATEGY: Randomised trials which included hydromorphone were sought using electronic databases and by handsearching relevant journals. Date of the most recent search: February 2000. SELECTION CRITERIA: RCTs which involved the administration of hydromorphone, for both acute and chronic pain conditions, in adults and children, were included. DATA COLLECTION AND ANALYSIS: A data extraction form was designed for the purpose of the review. The validity of each trial for inclusion was assessed using criteria described in the Cochrane Handbook. A grade was allocated to each study on the basis of allocation concealment. A checklist was used to assess blinding. MAIN RESULTS: Forty three studies (2725 subjects) were included in the review. Approximately half of these studies received a low quality score. In addition, the heterogeneity of the studies precluded combination of data and results. A meta-analysis was therefore not possible. Of the 43 included studies, 11 (645 subjects) involved chronic pain conditions (all cancer) and 32 (2080 subjects) acute pain. Three studies were placebo-controlled. Of the remainder, hydromorphone was compared with other opioids (morphine, fentanyl, sufentanyl, meperidine, oxycodone, diamorphine), bupivicaine and with itself, using different formulations. The routes of administration included intravenous, oral, spinal, intramuscular and subcutaneous. Overall, hydromorphone appears to be a potent analgesic. The limited number of studies available suggest that there is little difference between morphine and hydromorphone in terms of analgesic efficacy, adverse effect profile and patient preference. However, as most studies involved small numbers of patients, it is difficult to determine real differences between both drugs. In the context of both acute and chronic pain, the issue of equi analgesic ratios between morphine and hydromorphone was not resolved. REVIEWER'S CONCLUSIONS: The studies included in this review were varied in terms of quality and methodology. However, the majority demonstrated that hydromorphone is a potent analgesic, that the clinical effects of hydromorphone appear to be dose related, and that the adverse effect profile of hydromorphone is similar to that of other mu opioid receptor agonists. PMID- 11869662 TI - Non surgical interventions for late radiation proctitis in patients who have received radical radiotherapy to the pelvis. AB - BACKGROUND: Chronic radiation proctitis (inflammation of the rectum) may develop after the completion of pelvic radiotherapy. Presently there is no recommended standard management. OBJECTIVES: To assess the effects of various non-surgical treatment options for the management of late chronic radiation proctitis. SEARCH STRATEGY: We searched the Cochrane Controlled Trials Register, issue 1, 2001, MEDLINE 1966 to 2001, EMBASE 1980 to 2001, CANCERCD 1980 to 2001, Science Citation Index 1991 to 2001, CINAHL 1982 to 2001, as well as sources of grey literature. We also hand searched relevant textbooks and contacted experts in the field. SELECTION CRITERIA: Studies (preferentially randomised controlled trials) of interventions for the non-surgical management of late radiation proctitis in patients who have undergone pelvic radiotherapy as part of their cancer treatment. DATA COLLECTION AND ANALYSIS: The inclusion criteria were independently applied by two of the reviewers (AD and EJM) and where there was disagreement this was resolved by involving a third reviewer to form a consensus. MAIN RESULTS: Six randomised controlled trials were included. None of the trials compared anti-inflammatories with placebo. However rectal sucralfate showed greater clinical improvement for proctitis than anti-inflammatories (odds ratio 14.00, 95% confidence interval 1.46 to 134.26; n=1 study), though no difference was seen for endoscopic improvement (odds ratio 2.74, 95% confidence interval 0.64 to 11.76, n=1 study). The addition of metronidazole to the anti-inflammatory regime also appeared to improve the response rate, as measured by the reduction in rectal bleeding, diarrhoea, erythema and ulceration (n=1 study). Similarly rectal hydrocortisone appeared to be more effective than rectal betamethasone for clinical improvement although no difference was seen in endoscopic improvement (n=1 study). Short chain fatty acid enemas did not appear to be effective compared to placebo (n=2 studies). In the comparison of the heater probe and bipolar electrocautery (n=1 study), there was no discernible difference for severe bleeding after one year, but the heater probe demonstrated a greater increase in the haematocrit and reduced transfusion requirements. REVIEWER'S CONCLUSIONS: Late radiation complications are a relatively rare manifestation, with many potential carers and poor diagnostic criteria. Although certain interventions look promising and may be effective (such as rectal sucralfate, adding metronidazole to the anti-inflammatory regime and heater probes), single small studies (even if well conducted) provide insufficient evidence. The episodic and variable nature of late radiation proctitis also requires placebo controlled studies to establish whether particular treatments are effective. Regional or centralised registers of radiation toxicity should be established so that interventions can be administered in the setting of multi-centre trials with specific entry criteria, formal baseline and therapeutic assessments providing standardised outcome data including quality of life evaluations. PMID- 11869663 TI - Trypanocidal drugs for chronic asymptomatic Trypanosoma cruzi infection. AB - BACKGROUND: Prior guidelines stated that trypanocidal therapy should not be used for treating chronic asymptomatic Trypanosoma cruzi infections. However, the recent availability of clinical trials reporting high rates of parasitologic cure in children with early chronic T. cruzi infection have produced changes of these recommendations in some countries. Because of the uncertainty regarding best treatment for this stage of T. cruzi infections, the literature was reviewed systematically for a synthesis of the available evidence. OBJECTIVES: To assess the effects of trypanocidal therapy for chronic asymptomatic T. cruzi infection. SEARCH STRATEGY: We searched The Cochrane Controlled Trials Register (Issue 1, 2000), MEDLINE (start-Nov 1999), EMBASE (start - Feb 2000), LILACS (start - Feb 2000) and the Tropical Diseases Research Division of WHO database (Start - Feb 2000). Reference lists of articles were searched for relevant material. SELECTION CRITERIA: Published RCTs of trypanocidal therapy for people with chronic, asymptomatic T. cruzi infections DATA COLLECTION AND ANALYSIS: Two reviewers independently screened papers for inclusion criteria, quality assessment and data extraction. Forms were used to collect data. Reviewers resolved differences by discussion then a third reviewer if necessary. MAIN RESULTS: Of 43 papers assessed for inclusion, five RCTs (total population=756) met the inclusion criteria. The quality of the trials was rated as low (n=3) or intermediate (n=2). Two RCTs tested benznidazole in school children and three tested different agents in adults. The Odds Ratios and their 95%CI (Fixed models) were: Incidence of ECG abnormalities: 0.41 (0.09, 1.85); Negative seroconversion (AT ELISA): 10.91 (6.07, 19.58); Negative xenodiagnosis during the follow up: 5.37 (3.34, 8.64); Standardised mean reduction of antibody titres: 0.54 (0.31, 0.84). Nitroimidazolic derivatives substantially and significantly modified parasite related outcomes compared to placebo. Other agents showed borderline or not significant effect. REVIEWER'S CONCLUSIONS: Despite major public health importance, trypanocidal.therapy for chronic asymptomatic T. cruzi infection has been tested in few, small size RCTs which were designed to assess parasitic related, but not clinical outcomes. Therefore, the potential of trypanocidal therapy to prevent Chagas' disease among asymptomatic, chronically infected subjects is promising, but remains to be evaluated. trypanocidal therapy, particularly nitroimidazolic derivatives given to children or adults with positive xenodiagnosis improve parasite-related outcomes. The large contrast between the burden of Chagas disease and the existing evidence on its prevention points the need to test these or newer agents in more and larger RCTs that include clinical endpoints. PMID- 11869664 TI - Bisphosphonates for breast cancer. AB - BACKGROUND: Bone is the most common site of metastatic disease associated with breast cancer, and affects more than half of women during the course of their disease. Bone metastases are a significant cause of morbidity due to pain, pathological fractures, hypercalcaemia and spinal cord compression, and contribute to mortality. Bisphosphonates, which inhibit osteoclast-mediated bone resorption, are standard care for tumour-associated hypercalcaemia, and have been shown to reduce bone pain, improve quality of life, and to delay skeletal events and reduce their number in patients with multiple myeloma. Several randomized controlled trials have evaluated the role of bisphosphonates in breast cancer. OBJECTIVES: The aim of this systematic review was to identify, describe and summarize high-quality evidence regarding the effect of bisphosphonates on skeletal events, bone pain, quality of life and survival in women with early and advanced breast cancer. SEARCH STRATEGY: Randomized controlled trials were identified in the specialized register maintained by the secretariat of the Cochrane Breast Cancer Group (the search was applied to the databases Medline, Central/CCTR, Embase, CancerLit, and included handsearches from a number of other relevant sources). See: Cochrane Collaboration Collaborative Review Group in Breast Cancer search strategy. SELECTION CRITERIA: Randomized controlled trials evaluating skeletal events in women with metastatic breast cancer and in women with early breast cancer comparing: 1. treatment with a bisphosphonate with the same treatment without a bisphosphonate 2. treatment with one bisphosphonate with treatment with a different bisphosphonate. DATA COLLECTION AND ANALYSIS: Studies were selected by two independent reviewers. Studies fulfilling the eligibility criteria were evaluated for quality, particularly concealment of allocation to randomized groups. Data were extracted from the published papers or abstracts independently by the two primary reviewers for each of the specified endpoints (skeletal events, bone pain, quality of life and survival). Data on skeletal events and survival were presented as numbers of events, risk ratios and ratios of event rates. Meta-analyses were based on the fixed-effects model (Mantel Haenszel). Subjective qualitative ratings were used to summarize the quality of life and pain data. MAIN RESULTS: From 37 reports considered in detail after screening of the 117 reports identified by our search, 19 randomized studies were included. In eight studies that included 1962 women with advanced breast cancer and existing bone metastases, bisphosphonates reduced the risk of developing a skeletal event by 14% (RR 0.86; 95% confidence interval (CI) 0.80-0.91; P < 0.00001). This effect was more modest, but still highly significant if episodes of hypercalcaemia were excluded (6 studies, 1553 women, RR 0.88; 95% CI 0.81 0.96; P = 0.004). For intravenous pamidronate the reduction in the risk of skeletal event was greatest with a dosage of 90 mg (RR 0.77; 95% CI 0.69-0.87). Oral bisphosphonates reduced the risk of a skeletal event by 17% (pooled RR 0.83; 95% CI 0.73-0.94, P = 0.004). Oral clodronate reduced the risk of a skeletal event by 16% in women with advanced breast cancer and clinically evident bone metastases (pooled RR 0.84; 95% CI 0.72-0.98; P = 0.03). Compared with placebo or no bisphosphonate, with bisphosphonates the skeletal event rate was lower in all of eight studies (median reduction of 30%, range 20-48%); statistically significant reductions were reported in six trials (three intravenous pamidronate, two oral clodronate and one intravenous ibandronate). All studies of intravenous pamidronate and oral clodronate in women with advanced breast cancer and clinically evident bone metastases showed significant delays in the median time to a skeletal event. Event-free survival was reported to be longer in women receiving 6 mg of ibandronate compared with control women. Compared with placebo or no bisphosphonate, with bisphosphonates significant improvements in pain were reported in four studies, and improvements in quality of life were reported in two studies. Treatment with bisphosphonates does not appear to affect survival in women with advanced breast cancer. Intravenous zolendronate (4 mg) appeared to have equivalent efficacy when compared with intravenous pamidronate in a single randomized double-blind study. In the three studies of bisphosphonates in 320 women with advanced breast cancer without clinically evident bone metastases, there was no significant reduction in the incidence of skeletal events (RR 0.99; 95% CI 0.67-1.47; P > 0.9). In three studies of oral clodronate that included 1680 women with early breast cancer, there was borderline evidence of a reduction in the risk of developing skeletal metastases (RR 0.73; 95% CI 0.55-0.98; P = 0.04), but there was significant heterogeneity among these studies (P = 0.035). Toxicity or adverse events were described in 14 of the 19 studies. In general, few adverse events were reported. REVIEWER'S CONCLUSIONS: In women with advanced breast cancer and clinically evident bone metastases, the use of bisphosphonates (oral or intravenous) in addition to hormone therapy or chemotherapy, when compared with placebo or no bisphosphonates, reduces the risk of developing a skeletal event and the skeletal event rate, as well as increasing the time toskeletal event. Bisphosphonates may also reduce bone pain in women with advanced breast cancer and clinically evident bone metastases. In women with early breast cancer the effectiveness of oral clodronate in reducing the incidence of bone metastases remains an open question for research. PMID- 11869665 TI - Early versus deferred androgen suppression in the treatment of advanced prostatic cancer. AB - BACKGROUND: Prostate cancer is a leading cause of cancer death in men. Treatment goals for men with advanced prostate cancer include prolonging survival, preventing or delaying symptoms due to disease progression, improving and maintaining quality of life, reducing treatment related morbidity. Androgen suppression therapy is considered a mainstay of treatment for men with advanced prostate cancer. However it is not clear whether early androgen suppression for men with locally advanced disease or asymptomatic metastases improves length and quality of life compared to androgen suppression deferred until signs and symptoms of clinical progression. OBJECTIVES: This systematic review assessed the efficacy and adverse effects of primary therapy with early versus deferred androgen suppression therapy in men with advanced prostate cancer. SEARCH STRATEGY: Randomized controlled trials were searched in general and specialized databases (MEDLINE, EMBASE, CancerLIT, Cochrane Library, VA Cochrane Prostate Disease register) and by reviewing bibliographies including those of the Blue Cross and Blue Shield Association Technology Evaluation Center/Evidence-based Practice Center of the Agency for Healthcare Research and Quality (BCBS/TEC-AHRQ) report No.4. SELECTION CRITERIA: All published randomized trials were eligible for inclusion provided they: randomized men with advanced prostate cancer to early versus deferred androgen suppression; reported overall, progression-free, and cancer-specific survival, and/or adverse events; did not utilize androgen suppression as adjuvant therapy to radiation treatment. DATA COLLECTION AND ANALYSIS: An independent reviewer using a standardized form extracted information on trial characteristics, interventions, and outcomes. Results were reviewed for accuracy and discrepancies resolved by discussion. The main outcome measure for comparing effectiveness was the overall survival at 1, 2, 5 and 10 years. Progression-free survival, cancer-specific survival, complications due to disease progression and the incidence of adverse effects of treatment were also measured. MAIN RESULTS: Four trials involving 2,167 patients were included in this review. All of the trials were conducted prior to use of prostate specific antigen (PSA) testing. There was variability between studies regarding the treatments used and the requirements for initiation of treatments. The percent overall survival at 1, 2, 5, and 10 years for the early treatment group was 88%, 73%, 44%, and 18%. For the deferred therapy group the percent overall survival was 86%, 71%, 37%, and 12%. The pooled estimate for the difference in overall survival favored early therapy but was significant only at 10 years when few patients had survived [OR = 1.16 (95% CI: 0.90 to 1.49) at 1 year, 1.08 (95% CI: 0.89 to 1.33) at 2 years, 1.19 (95% CI: 0.95 to 1.50) at 5 years, and 1.50 (95% CI: 1.04 to 2.16) at 10 years]. The pooled estimate of prostate cancer specific survival at 2, 5, and 10 years favored early therapy though the confidence intervals were wide and the results not statistically different. The risk differences at 2, 5, and 10 years were 2.7%, 5.8%, and 4.6%. Although each study used unique definitions of progression free survival, all studies found progression free survival was consistently better in the early intervention group at all time points. Complications due to disease progression were only reported in one study but were more frequent in the deferred treatment group. Adverse events due to treatment were also only reported in one study but occurred more frequently in the early treatment group. REVIEWER'S CONCLUSIONS: The evidence from randomized controlled trials is limited by the variability in study design, stage of cancer and subjects enrolled, interventions utilized, definitions and reporting of outcomes and the lack of PSA testing for diagnostic and monitoring purposes. However, the available information suggests that early androgen suppression for treatment of advanced prostate cancer reduces disease progression and complications due to progression. Early androgen suppression may provide a small but statistically significant improvement in overall survival at 10 years. There was no statistically significant difference in prostate cancer specific survival but a clinically important difference could not be excluded. These outcomes need to be evaluated with the evidence suggesting higher costs and more frequent treatment related adverse effects with early therapy. Additional studies are required to evaluate more definitively the efficacy and adverse effects of early versus delayed androgen suppression in men with prostate cancer. In particular trials should evaluate patients with advanced prostate cancer diagnosed by PSA testing and men with persistent or rising PSA levels following treatment options (e.g. radical prostatectomy, radiation therapy or observation) for clinically localized disease. PMID- 11869666 TI - Antiretrovirals for reducing the risk of mother-to-child transmission of HIV infection. AB - BACKGROUND: At the end of 2000 it was estimated that over 36 million people were living with the human immunodeficiency virus (HIV). This includes 1.4 million children less than 15 years of age. This is one of several reviews assessing the available evidence for preventing mother-to-child transmission of HIV infection. The other reviews will address other interventions, including Caesarean section, breast feeding, vaginal lavage and vitamin A supplementation. OBJECTIVES: To assess which antiretroviral therapies may be effective in decreasing the risk of mother-to-child transmission of HIV infection as well as their effect on neonatal and maternal mortality and morbidity. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register. We also searched conference abstracts from the International AIDS Conferences and Conference on Retroviruses and Opportunistic Infections. SELECTION CRITERIA: Randomised trials comparing any antiretroviral therapy aimed at decreasing the risk of mother-to-child transmission of HIV infection with placebo or no treatment, or any two or more antiretroviral therapies or regimens aimed at decreasing the risk of mother-to-child transmission of HIV infection. DATA COLLECTION AND ANALYSIS: Two reviewers independently extracted data and assessed trial quality. MAIN RESULTS: Zidovudine monotherapy Any zidovudine regimen versus placebo significantly reduces the risk of mother-to-child transmission (Peto odds ratio (OR) 0.46, 95% confidence interval (CI) 0.35 to 0.60). Zidovudine also appears to decrease the risk of infant death within the first year of birth (OR 0.57, 95% CI 0.38 to 0.85) and the risk of maternal death (OR 0.32, 95% CI 0.16 to 0.66). There is no evidence that zidovudine influences the incidence of premature delivery (OR 0.86, 95% CI 0.57 to 1.29) or low birth weight (OR 0.74, 95% CI 0.53 to 1.04). The risk of transmission using a 'short short' course of zidovudine (from 35 weeks in pregnancy for the mother and for the baby until 3 days old) was higher than the risk using a 'long-long' course (from 28 weeks in pregnancy for the mother and for the baby until 6 weeks old), (OR 2.55, 95% CI 1.26 to 5.18). However, the effectiveness of the 'long-short' course (from 28 weeks in pregnancy for the mother and for the baby until 3 days old) and the 'short-long' course (from 35 weeks in pregnancy for the mother and for the baby until 6 weeks old) did not differ from that of the 'long-long' course. Nevirapine One large randomised controlled trial demonstrates that nevirapine given to mothers as a single dose at the onset of labour and to babies as a single dose within 72 hours of birth is more effective than an intrapartum and post-partum regimen of zidovudine (OR 0.51, 95% CI 0.33 to 0.79). When nevirapine is given to mothers already receiving standard antiretroviral therapy, however, there appears to be no additional advantage (OR 1.10, 95% CI 0.42 to 2.86). Combination Therapy Preliminary findings of the effect of combination therapy using zidovudine and lamivudine (3TC) suggest a decrease in the risk of transmission when the combination is given during the antenatal and intrapartum period or during the intrapartum and postpartum period compared with placebo. There is no evidence that intrapartum zidovudine and lamivudine alone are sufficient to decrease the risk of transmission compared with placebo. REVIEWER'S CONCLUSIONS: Implications for practice The randomised trials included in this review provide evidence that short course zidovudine and single-dose nevirapine are effective therapies for reducing mother-to-child transmission of HIV. The challenge for low and middle income countries will be to institute this therapy in practice. In industrialised countries practice has already moved on from the current evidence and combination antiretroviral therapy aimed primarily at preventing disease progression in the mother is the standard of care. Implications for research The potential value of nevirapine used for longer durations in breastfeeding populations should be considered as it may further reduce the risk of mother-to-child transmission, particularly if combined with early weaning. On-going evaluation of combination antiretroviral therapy is essential and will have an immediate benefit for countries with the resources to adopt such treatment. The search for effective, affordable, safe and acceptable alternatives to antiretroviral therapy for reducing mother-to-child transmission in resource poor countries should remain on the research agenda. PMID- 11869667 TI - Optimal duration of exclusive breastfeeding. AB - BACKGROUND: : Although the health benefits of breastfeeding are widely acknowledged, opinions and recommendations are strongly divided on the optimal duration of exclusive breastfeeding. Much of the debate has centered on the so called 'weanling's dilemma' in developing countries: the choice between the known protective effect of exclusive breastfeeding against infectious morbidity and the (theoretical) insufficiency of breast milk alone to satisfy the infant's energy and micronutrient requirements beyond four months of age. The debate over whether to recommend exclusive breastfeeding for four to six months versus 'about six months' has recently become heated and acrimonious. OBJECTIVES: : The primary objective of this review was to assess the effects on child health, growth, and development, and on maternal health, of exclusive breastfeeding for six months versus exclusive breastfeeding for three to four months with mixed breastfeeding (introduction of complementary liquid or solid foods with continued breastfeeding) thereafter through six months. A secondary objective was to assess the child and maternal health effects of prolonged (greater than six months) exclusive breastfeeding versus exclusive breastfeeding for six months followed by mixed breastfeeding thereafter. SEARCH STRATEGY: : Two independent literature searches were carried out, together comprising the following databases: MEDLINE (as of 1966), Index Medicus (prior to 1966), CINAHL, HealthSTAR, BIOSIS, CAB Abstracts, EMBASE-Medicine, EMBASE-Psychology, Econlit, Index Medicus for the WHO Eastern Mediterranean Region, African Index Medicus, Lilacs (Latin American and Caribbean literature), EBM Reviews-Best Evidence, the Cochrane Database of Systematic Reviews (The Cochrane Library Issue 3, 2000), and the Cochrane Controlled Trials Register (The Cochrane Library Issue 3, 2000). No language restrictions were imposed. The two searches yielded a total of 2,668 unique citations. Contacts with experts in the field yielded additional published and unpublished studies. SELECTION CRITERIA: : We selected all internally-controlled clinical trials and observational studies comparing child or maternal health outcomes with exclusive breastfeeding for six or more months versus exclusive breastfeeding for at least three to four months with continued mixed breastfeeding until at least six months. Studies were stratified according to study design (controlled trials versus observational studies), provenance (developing versus developed countries), and timing of compared feeding groups (three to seven months versus later). DATA COLLECTION AND ANALYSIS: : Two reviewers independently assessed study quality (using a priori assessment criteria) and extracted data. MAIN RESULTS: : Twenty independent studies meeting the selection criteria were identified by the literature search: nine from developing countries (two of which were controlled trials in Honduras) and 11 from developed countries (all observational studies). The two trials did not receive high methodologic quality ratings but were nonetheless superior to any of the observational studies included in this review. The observational studies were of variable quality; in addition, their nonexperimental designs were not able to exclude potential sources of confounding and selection bias. Definitions of exclusive breastfeeding varied considerably across studies. Neither the trials nor the observational studies suggest that infants who continue to be exclusively breastfed for six months show deficits in weight or length gain, although larger sample sizes would be required to rule out modest differences in risk of undernutrition. The data are conflicting with respect to iron status, but at least in developing country settings where newborn iron stores may be suboptimal, suggest that exclusive breastfeeding without iron supplementation through six months may compromise hematologic status. Based primarily on an observational analysis of a large randomized trial in Belarus, infants who continue exclusive breastfeeding for six months or more appear to have a significantly reduced risk of one or more episodes of gastrointestinal infection. No significant reduction in risk of atopic eczema, asthma, or other atopic outcomes has been demonstrated in studies from Finland, Australia, and Belarus. Data from the two Honduran trials suggest that exclusive breastfeeding through six months is associated with delayed resumption of menses and more rapid postpartum weight loss in the mother. REVIEWER'S CONCLUSIONS: : We found no objective evidence of a 'weanling's dilemma'. Infants who are exclusively breastfed for six months experience less morbidity from gastrointestinal infection than those who are mixed breastfed as of three or four months, and no deficits have been demonstrated in growth among infants from either developing or developed countries who are exclusively breastfed for six months or longer. Moreover, the mothers of such infants have more prolonged lactational amenorrhea. Although infants should still be managed individually so that insufficient growth or other adverse outcomes are not ignored and appropriate interventions are provided, the available evidence demonstrates no apparent risks in recommending, as a general policy, exclusive breastfeeding for the first six months of life in both developing and developed country settings. Large randomized trials are recommended in both types of setting to rule out small effects on growth and to confirm the reported health benefits of exclusive breastfeeding for six months or beyond. PMID- 11869668 TI - Electromagnetic fields for the treatment of osteoarthritis. AB - BACKGROUND: As the focus for osteoarthritis (OA) treatment shifts away from drug therapy, we consider the effectiveness of pulsed electric stimulation which is proven to stimulate cartilage growth on the cellular level. OBJECTIVES: 1)To assess the effectiveness of pulsed electric stimulation for the treatment of osteoarthritis (OA). 2) To assess the most effective and efficient method of applying an electromagnetic field, through pulsed electromagnetic fields (PEMF) or electric stimulation, as well as the consideration of length of treatment, dosage, and the frequency of the applications. SEARCH STRATEGY: We searched PREMEDLINE, MEDLINE, HealthSTAR, CINAHL, PEDro, and the Cochrane Controlled Trials Register (CCTR) up to and including 2001. This included searches through the coordinating offices of the trials registries of the Cochrane Field of Physical and Related Therapies and the Cochrane Musculoskeletal Group for further published and unpublished articles. The electronic search was complemented by hand searches and experts in the area. SELECTION CRITERIA: Randomized controlled trials and controlled clinical trials that compared PEMF or direct electric stimulation against placebo in patients with OA. DATA COLLECTION AND ANALYSIS: Two reviewers determined the studies to be included in the review based on inclusion and exclusion criteria (JH,VR) and extracted the data using pre developed extraction forms for the Cochrane Musculoskeletal Group. The methodological quality of the trials was assessed by the same reviewers using a validated scale (Jadad 1996). Osteoarthritis outcome measures were extracted from the publications according to OMERACT guidelines (Bellamy 1997) and additional secondary outcomes considered. MAIN RESULTS: Only three studies with a total of 259 OA patients were included in the review. Electrical stimulation therapy had a small to moderate effect on outcomes for knee OA, all statistically significant with clinical benefit ranging from 13-23% greater with active treatment than with placebo. Only 2 outcomes for cervical OA were significantly different with PEMF treatment and no clinical benefit can be reported with changes of 12% or less. REVIEWER'S CONCLUSIONS: Current evidence suggests that electrical stimulation therapy may provide significant improvements for knee OA, but further studies are required to confirm whether the statistically significant results shown in these trials confer to important benefits. PMID- 11869669 TI - Shock wave therapy for lateral elbow pain. AB - BACKGROUND: This review is one in a series of reviews of interventions for lateral elbow pain. Lateral elbow pain, or tennis elbow, is a common condition causing pain in the elbow and forearm and lack of strength and function of the elbow and wrist. Shock wave therapy (ESWT) involves the application of single pulsed acoustic wave. Since the 1990's reports of benefit of ESWT in the treatment of tendon disorders have been appearing in the literature. A systematic review published in the German language appeared in 2000 (Boddeker 2000) OBJECTIVES: To determine the effectiveness and safety of ESWT in the treatment of adults with lateral elbow pain. SEARCH STRATEGY: Comprehensive electronic searches of MEDLINE, CINAHL, EMBASE and SCISEARCH were combined with searches of the Cochrane Clinical Trails Registrar and the Musculoskeletal Review Group's specialist trial database. Identified keywords and authors were searched again in an effort to identify as many trials as possible. SELECTION CRITERIA: Two independent reviewers assessed all identified trials against pre-determined inclusion criteria. Randomised and pseudo randomised trials in all languages were evaluated for inclusion in the review provided they described individuals with lateral elbow pain and were comparing the use of ESWT as a treatment strategy. DATA COLLECTION AND ANALYSIS: For continuous variables means and standard deviations were extracted or imputed to allow the analysis of weighted mean difference. Weighted mean difference using a random effects model was selected when outcomes were measured on standard scales. A fixed effects model was used to interpret results and assess heterogeneity. For binary data numbers of events and total population were analysed and interpreted as relative risk. MAIN RESULTS: Two trials of ESWT versus placebo are included in this review (~~Rompe 1996~~, ~~Haake 2001~~). Both trials included similar study populations consisting of participants with chronic symptoms who had failed other conservative treatment. The frequency of ESWT application and the doses and techniques used were similar in both trials. The first trial demonstrated highly significant differences in favour of ESWT whereas the second trial found no benefits of ESWT over placebo. When the data from the two trials were pooled, the benefits observed in the first trial were no longer statistically significant. The relative risk for treatment failure (defined as Roles-Maudsley score of 4) of ESWT over placebo was 0.40 (95% CI, 0.08 to 1.91) at six weeks and 0.44 (95% CI, 0.09 to 2.17) at one year. After 6 weeks, there was no statistically significant improvement in pain at rest [WMD pain out of 100 = - 11.40 (95% CI, -26.10 to 3.30)], pain with resisted wrist extension [WMD pain out of 100 = -16.20 (95% CI, -47.75 to 15.36)] or pain with resisted middle finger extension [WMD pain out of 100 = -20.51(95% CI, -56.57 to 15.56)]. Results after 12 or 24 weeks were similar. REVIEWER'S CONCLUSIONS: The two trials included in this review yielded conflicting results. Further trials are needed to clarify the value of ESWT for lateral elbow pain. PMID- 11869670 TI - Surgery for lateral elbow pain. AB - BACKGROUND: This review is one in a series of reviews of interventions for lateral elbow pain. Lateral elbow pain, or tennis elbow, is a common condition causing pain in the lateral elbow and forearm and lack of strength and function of the elbow and wrist. Surgery is sometimes recommended in treating chronic cases of lateral elbow pain where other less invasive interventions have failed. Various operations have been described based upon the surgeon's concept of the pathological entity. The most described surgical procedures involve release of the extensor carpi radialis brevis (ECRB) from the lateral epicondyle region based upon the premise that there is pathology in the attachment of ECRB to the lateral epicondyle. No systematic review has previously been published assessing the effect of surgical interventions for lateral elbow pain. OBJECTIVES: To determine the effectiveness of surgical interventions in the treatment of adults with lateral elbow pain. SEARCH STRATEGY: Comprehensive electronic searches of MEDLINE, CINAHL, EMBASE and SCISEARCH were combined with searches of the Cochrane Clinical Trials Registrar and the Musculoskeletal Review Group's specialist trial database. Identified keywords and authors were researched in an effort to identify as many trials as possible. Searches were conducted up to October 2001. SELECTION CRITERIA: Two independent reviewers assessed all identified studies against pre-determined inclusion criteria. Randomised and pseudo randomised trials in all languages were to be included in the review provided they were studying the effects of a surgical intervention and included a control as treatment for adults with lateral elbow pain. The control intervention could comprise no treatment or another intervention including an alternate surgical intervention. Outcomes of interest included pain, function, disability and quality of life, strength and adverse effects. DATA COLLECTION AND ANALYSIS: The planned collection and analysis of data is described. MAIN RESULTS: Our search did not identify any controlled trials investigating the effect of surgery on lateral elbow pain. REVIEWER'S CONCLUSIONS: At this time there are no published controlled trials of surgery for lateral elbow pain. Without a control group, it is not possible to draw any conclusions about the value of this modality of treatment. PMID- 11869671 TI - Acupuncture for lateral elbow pain. AB - BACKGROUND: This review is one in a series of reviews of interventions for lateral elbow pain. Lateral elbow pain, or tennis elbow, is a common condition causing pain in the elbow and forearm and lack of strength and function of the elbow and wrist. Acupuncture has long been used to treat lateral elbow pain in China and in Western countries practitioners and consumers are increasingly exploring acupuncture as a first line treatment for musculoskeletal disorders. No previous systematic review of the available evidence has been conducted to determine whether acupuncture is efficacious in the treatment lateral elbow pain. OBJECTIVES: To determine the effectiveness of acupuncture in the treatment of adults with lateral elbow pain with respect to pain reduction, improvement in function, grip strength and adverse effects. SEARCH STRATEGY: We searched MEDLINE, CINAHL, EMBASE and SCISEARCH and the Cochrane Clinical Trials Register and the Musculoskeletal Review Group's specialist trial database from 1966 to June 2001. Identified keywords and authors were searched in an effort to retrieve as many trials as possible. SELECTION CRITERIA: Two independent reviewers assessed all identified trials against pre-determined inclusion criteria. Randomised and pseudo randomised trials in all languages were included in the review provided they were testing acupuncture compared to placebo or another intervention in adults with lateral elbow pain (tennis elbow). Outcomes of interest were pain, function, disability, quality of life, strength, participant satisfaction with treatment and adverse effect. DATA COLLECTION AND ANALYSIS: For continuous variables means and standard deviations were extracted or imputed to allow the analysis of weighted mean difference, while for binary data numbers of events and total population were analysed and interpreted as relative risks. Trial results were combined only in the absence of clinical and statistical heterogeneity. MAIN RESULTS: Four small randomized controlled trials were included but due to flaws in study designs (particularly small populations, uncertain allocation concealment and substantial loss to follow up) and clinical differences between trials, data from trials could not be combined in a meta analysis. One randomised controlled trial found that needle acupuncture results in relief of pain for significantly longer than placebo (WMD = 18.8 hours, 95%CI 10.1 to 27.5) and is more likely to result in a 50% or greater reduction in pain after 1 treatment (RR 0.33, 95%CI 0.16 to 0.69) (Molsberger 1994). A second randomized controlled trial demonstrated needle acupuncture to be more likely to result in overall participant reported improvement than placebo in the short term (RR = 0.09 95% CI 0.01 to 0.64) (Haker 1990a). No significant differences were found in the longer term (after 3 or 12 months). A randomized controlled trial of laser acupuncture versus placebo demonstrated no differences between laser acupuncture and placebo with respect to overall benefit (Haker 1990b). A fourth included trial published in Chinese demonstrated no difference between Vitamin B12 injection plus acupuncture, and Vitamin B12 injection alone (Wang 1997). REVIEWER'S CONCLUSIONS: There is insufficient evidence to either support or refute the use of acupuncture (either needle or laser) in the treatment of lateral elbow pain. This review has demonstrated needle acupuncture to be of short term benefit with respect to pain, but this finding is based on the results of 2 small trials, the results of which were not able to be combined in meta analysis. No benefit lasting more than 24 hours following treatment has been demonstrated. No trial assessed or commented on potential adverse effect. Further trials, utilising appropriate methods and adequate sample sizes, are needed before conclusions can be drawn regarding the effect of acupuncture on tennis elbow. PMID- 11869672 TI - Deep transverse friction massage for treating tendinitis. AB - BACKGROUND: Deep transverse friction massage (DTFM) is one of several physiotherapy interventions suggested for the management of pain due to iliotibial band friction syndrome (ITBFS). OBJECTIVES: To assess the effectiveness of DTFM for treating ITBFS observed in runners. SEARCH STRATEGY: We searched the Medline, Embase, Healthstar, Sports Discus, CINAHL, the Cochrane Controlled Trials Register, PEDro, the specialized registry of the Cochrane musculoskeletal group and the Cochrane field of Physical and Related Therapies up to the end of December 2000, using the sensitive search strategy developed by the Cochrane Collaboration. The search was complemented with bibliography searching of the reference list of the trials retrieved from the electronic search. Key experts in the area were contacted for further published and unpublished articles. SELECTION CRITERIA: All randomized controlled trials (RCTs) and controlled clinical trials (CCTs) comparing therapeutic ultrasound against placebo or another active intervention in patients with patellofemoral pain syndrome were selected. DATA COLLECTION AND ANALYSIS: Two reviewers determined the studies to be included based on inclusion and exclusion criteria (LB, VR). Data were independently abstracted by two reviewers (VR, LB), and checked by a third reviewer (BS) using a pre-developed form of the Cochrane Musculoskeletal Group. The same two reviewers, using a validated scale, assessed the methodological quality of the RCTs and CCTs independently. Iliotibial band friction syndrome outcome measures were extracted from the publications. The pooled analysis was performed using weighted mean differences (WMDs) for pain relief as described as 1) daily pain; 2) pain while running and 3) percentage of maximum pain when running. A chi-square test was used to assess heterogeneity among trials. Fixed effects models were used throughout and random effects for outcomes showing heterogeneity. MAIN RESULTS: One RCT, including 17 patients with ITBFS was included. The experimental group (DTFM combined to rest, stretching exercises, cryotherapy and therapeutic ultrasound) (n=9) was compared to the control group (rest, stretching exercises, cryotherapy and therapeutic ultrasound only) (n=8). This trial showed no statistical difference in the three types of pain relief measured after four consecutive sessions of DTFM combined with other physiotherapy modalities for runners. Despite lack of statistical significance, there was a clinically important relative percentage difference in pain while running of 22%. REVIEWER'S CONCLUSIONS: DTFM combined with other physiotherapy modalities did not show consistent benefit over control of pain for runners experiencing ITBFS. These conclusions are limited by the low methodological quality of the one small sample size trial (n=17) included. No conclusions can be drawn about the use or non use of DTFM for the treatment of ITBFS. Future trials, utilizing appropriate methods and adequate sample sizes are needed before conclusions can be drawn regarding the effect of massage on iliotibial band friction syndrome. PMID- 11869673 TI - Inhaled beclomethasone versus budesonide for chronic asthma. AB - BACKGROUND: Beclomethasone dipropionate (BDP) and budesonide (BUD) are used widely in the treatment of chronic asthma. The two drugs have different in vitro pharmacokinetic characteristics. It is unclear whether this translates into clinically significant differences in efficacy or safety when treating children and adults with chronic asthma. OBJECTIVES: To assess clinical outcomes in studies which have compared inhaled BDP and BUD in the treatment of chronic asthma. SEARCH STRATEGY: We searched the Cochrane Airways Group Trial Register (1999) and reference lists of articles. We contacted trialists and pharmaceutical companies for additional studies and searched abstracts of major respiratory society meetings (1997-1999). SELECTION CRITERIA: Prospective, randomised trials comparing BDP to BUD in the treatment of chronic asthma. Two reviewers independently assessed articles for inclusion and methodological quality. DATA COLLECTION AND ANALYSIS: One reviewer extracted data; authors were contacted to clarify missing information. Quantitative analyses where undertaken using Review Manager 4.0.3 with Metaview 3.1. MAIN RESULTS: 24 studies met the criteria for inclusion (1174 subjects). Methodological quality was variable. A meta-analysis of crossover studies did not demonstrate a significant difference between BDP and BUD for FEV1, morning PEF, evening PEF, asthma symptoms or rescue beta2 agonist use, over a dose range of 400 to 1000 mcg/d. The majority of crossover trials had significant design flaws related to a lack of washout and/or failure to exclude carryover effects so the results must be viewed with caution. A single crossover study with adequate washout showed that BUD 400 mcg/d delivered via Turbohaler dry powder inhaler (DPI) may be more effective than BDP 400 mcg/d delivered via Rotahaler DPI in reducing histamine bronchial hyper-responsiveness: Weighted Mean Difference (WMD) 0.43 log10 PC20 FEV1 (95% Confidence Intervals (CI) 0.05, 0.81 log10 PC20 FEV1). A meta-analysis of two parallel group, dose down-titration studies (231 patients) showed that less BUD delivered via a Turbohaler DPI was required to maintain control in adults asthmatics compared to BDP delivered via metered dose inhaler with or without a spacer: WMD 444 mcg/d (95% CI 332, 556 mcg/d). REVIEWER'S CONCLUSIONS: There is limited high quality randomised controlled trial data comparing the relative efficacy of BDP and BUD. Current guidelines (BTS 1997, GINA 1995, NHLBI 1997) assume BDP and BUD to have equal efficacy, such that for each defined level of asthma severity, the recommended doses BDP and BUD are the same. Although there is some data to suggest that BUD via Turbohaler is more effective than BDP via either Rotahaler or MDI (with and without spacer), these comparisons are confounded by use of different delivery devices, and are not sufficient to warrant a change in guideline recommendations. PMID- 11869674 TI - Primary care based clinics for asthma. AB - BACKGROUND: Primary care clinics for asthma have been encouraged and are becoming widespread in some countries, particularly in the UK. OBJECTIVES: To determine the effectiveness of organised asthma care via primary care based asthma clinics. SEARCH STRATEGY: A search of the Cochrane Airways Group register and Cochrane Controlled Trials Register using the following search strategy: clinic* OR general pract* OR family pract* or primary care. Separate and additional searches were also conducted using MEDLINE, CINAHL and EMBASE databases. SELECTION CRITERIA: Trials had to be performed in primary care and be restricted to patients with asthma. Care could be delivered by doctor or nurse. Two reviewers independently ascertained the relevance of trials from titles and abstracts obtained from the searches. Relevant full text articles were retrieved with two reviewers assessing each study for inclusion. DATA COLLECTION AND ANALYSIS: Two reviewers independently conducted all data abstraction and analysis and all disagreements were resolved by discussion. For the dichotomous variables, odds ratio (OR) or relative risks (RR) with 95% Confidence Interval (95%CI) were calculated for individual outcomes. MAIN RESULTS: Only one trial met the criteria for inclusion in the review. This trial provided 11 outcome measures of which two showed a significant effect of the intervention. More patients in the intervention group had peak flow meters (RR 1.30; 95%CI 1.05,1.61) and fewer patients in the intervention group were likely to wake up at nights due to their asthma (RR 0.30; 95%CI 0.16, 0.81). REVIEWER'S CONCLUSIONS: There is limited evidence of benefit for primary care based asthma clinics, but firm conclusions cannot be formed until more good quality trials have been carried out. PMID- 11869675 TI - Inhaled fluticasone at different doses for chronic asthma. AB - BACKGROUND: Inhaled fluticasone propionate (FP) is a high potency inhaled corticosteroid used in the treatment of asthma. OBJECTIVES: 1. To assess the efficacy and safety outcomes in studies that compared inhaled fluticasone at different nominal daily doses in the treatment of chronic asthma. 2. To test for the presence of a dose response effect. SEARCH STRATEGY: We searched the Cochrane Airways Group Trial Register (1999) and reference lists of articles. We contacted trialists and pharmaceutical companies for additional studies and searched abstracts of major respiratory society meetings (1997-1999). SELECTION CRITERIA: Randomised trials in children and adults comparing fluticasone at different nominal daily doses in the treatment of chronic asthma. Two reviewers independently assessed articles for inclusion and methodological quality. DATA COLLECTION AND ANALYSIS: One reviewer extracted data. Quantitative analyses where undertaken using Review Manager 4.0.3 with Metaview 3.1. MAIN RESULTS: 20 studies (>6000 patients) met the inclusion criteria. Methodological quality was high. In non-oral steroid treated asthmatics with mild-moderate disease, a dose response effect was present for morning PEF; when comparing low doses (200 vs 100 mcg/d) Weighted Mean Difference (WMD) 6 L/min, 95% Confidence Interval (CI) 1, 10 L/min; medium-low dose (400-500 vs 100 mcg/d), WMD 8 L/min, 95%CI 1,15 L/min); and high vs low dose (800-1000 vs 50-100 mcg/d), WMD 22 L/min 95% CI 15,29 L/min). There was no dose response in symptoms or rescue beta2 agonist use. Hoarseness and oral Candidiasis was significantly higher with 800-1000 mcg/d than 50-100 mcg/d. In oral steroid dependent disease gain 2000 mcg/d had a greater effect than 1000 1500 mcg/d in likelihood of stopping prednisolone (Peto Odds Ratio 2.8, 95% CI 1.3, 6.3) and reduced daily prednisolone dose (WMD 2.0 mg/d, 95% CI 0.1, 4.0 mg/d). REVIEWER'S CONCLUSIONS: Effects of fluticasone are dose dependent but relatively small. Patients with mild to moderate disease achieve similar levels of asthma control on low doses of fluticasone (200 mcg/d or less) as they do on high doses (500 mcg/d or greater). In oral corticosteroid dependent asthmatics, reductions in prednisolone requirement may be gained with FP 2000 mcg/d. PMID- 11869676 TI - Inhaled corticosteroid effects on bone metabolism in asthma and mild chronic obstructive pulmonary disease. AB - BACKGROUND: Inhaled corticosteroids form the main therapy for asthma, but there is increasing concern about the potential systematic effects of long-term inhaled corticosteroids including their effect on bone metabolism and bone loss. OBJECTIVES: To determine the effect of inhaled corticosteroids use on biochemical markers of bone turnover, bone mineral density and the development of fractures. SEARCH STRATEGY: We searched the Cochrane Airways Group trials register, electronic reference databases, UK National Research Register, bibliographies of included studies, and contacted pharmaceutical companies. SELECTION CRITERIA: Randomised trials of the effect of inhaled steroid versus placebo on markers of bone function and metabolism, in adults with asthma or mild COPD. DATA COLLECTION AND ANALYSIS: Trial quality was assessed and data extracted from the papers included (2 reviewers per paper) and from additional data supplied by the authors. MAIN RESULTS: Of 438 references found, seven met the inclusion criteria. Three studies were in healthy subjects asthma or COPD. The patients were generally less than 60 years old and the male:female ratio was 2:1. There was no evidence of increased risk of loss of bone mineral density (BMD) or fractures. There was no significant change in osteocalcin at conventional doses of inhaled corticosteroids (Standardised Mean Difference [SMD] -0.34 (95% Confidence Interval [CI] -0.72, 0.04), although a statistically significant change was seen in those studies using experimental doses of inhaled steroid in excess of the doses recommended by the British Thoracic Society SMD 0.97 (95% CI -1.61, -0.34). A statistically significant change in parathyroid hormone seen in one small short trial (n=10, 6 weeks) may have been due to the trial design and outcome measurements used. REVIEWER'S CONCLUSIONS: In patients with asthma or mild COPD, there is no evidence of an effect of inhaled corticosteroid at conventional doses given for two or three years on BMD or vertebral fracture. Higher doses were associated with biochemical markers of increased bone turnover, but data on BMD and fractures at these doses are not available. There is a need for further, even longer term prospective studies of conventional and high doses of inhaled corticosteroids. PMID- 11869677 TI - Institutional versus at-home long term care for functionally dependent older people. AB - BACKGROUND: An increasing number of functionally dependent older people require care and medical treatment. Increasingly governments are shifting resources into community care expecting both reduction in costs and improvement in the quality of care. However, it is difficult to establish the costs and benefits of institutional and the alternative at-home care. OBJECTIVES: To assess the effects of institutional versus at-home care for functionally dependent older people on health outcomes, satisfaction (of functionally dependent older people, relatives and health care professionals), quality of care and costs. SEARCH STRATEGY: We searched the Cochrane Effective Practice and Organisation of Care Group (EPOC) specialised register (1999), the Cochrane Controlled Trials Register (Issue 4 1999), MEDLINE (1966 to 1999), EMBASE (1980 to 1999), Best Evidence (1990 to 1999), Ageline (1982 to 1999), Cinahl (1982 to 1999), EconLit (1969 to 1999), PsycInfo (1887 to 1999), NTIS (1980 to 1999), Scisearch (1980 to 1999), Sigle (1980 to 1999), and reference lists of related systematic reviews and articles. We contacted authors working in the field in an attempt to identify unpublished studies. SELECTION CRITERIA: Randomised trials, controlled clinical trials, controlled before and after studies and interrupted time series studies where functionally dependent older people were assigned to either institutional or at home care. DATA COLLECTION AND ANALYSIS: Three reviewers independently extracted data and assessed study quality. No meta analysis was conducted as only one trial was identified. MAIN RESULTS: One study was included involving 112 people. This evaluated a community care programme (CCP) organising foster care versus nursing home care. No studies were found where functionally dependent older people returned to their own homes. The included trial was small and of poor methodological quality. No significant difference was found between the two groups when comparing functioning (ADL and IADL), mental status, attitudes to perceived health, life satisfaction or mortality. REVIEWER'S CONCLUSIONS: There is insufficient evidence to estimate the likely benefits, harms and costs of institutional or at-home care for functionally dependent older people. PMID- 11869678 TI - Arthritis critically dependent on innate immune system players. AB - K/BxN T cell receptor transgenic mice are a model of inflammatory arthritis, similar to rheumatoid arthritis. Disease in these animals is focused specifically on the joints but stems from autoreactivity to a ubiquitously expressed antigen, glucose-6-phosphate isomerase (GPI). T and B cells are both required for disease initiation, but anti-GPI immunoglobulins (Igs), alone, can induce arthritis in lymphocyte-deficient recipients. Here, we show that the arthritogenic Igs act through both Fc receptors (in particular, FcgammaRIII) and the complement network (C5a). Surprisingly, the alternative pathway of complement activation is critical, while classical pathway components are entirely dispensable. We suggest that autoimmune disease, even one that is organ specific, can occur when mobilization of an adaptive immune response results in runaway activation of the innate response. PMID- 11869679 TI - Tolerance to islet antigens and prevention from diabetes induced by limited apoptosis of pancreatic beta cells. AB - Crosspresentation of self-antigens by antigen-presenting cells is critical for the induction of peripheral tolerance. As apoptosis facilitates the entry of antigens into the crosspresentation pathway, we sought to prevent the development of autoimmune diabetes by inducing pancreatic beta cell apoptosis before disease onset. Accordingly, young nonobese diabetic (NOD) mice injected with a single low dose of streptozotocin (SZ), a drug cytotoxic for beta cells, exhibited impaired T cell responses to islet antigens and were protected from spontaneous diabetes. Furthermore, beta cell apoptosis was necessary for protection since SZ did not protect RIP-CrmA transgenic NOD mice in which beta cells expressed the caspase inhibitor CrmA. Our results support a model in which apoptosis of pancreatic beta cells induces the development of regulatory cells leading to the tolerization of self-reactive T cells and protection from diabetes. PMID- 11869680 TI - Pancreatic lymph node-derived CD4(+)CD25(+) Treg cells: highly potent regulators of diabetes that require TRANCE-RANK signals. AB - Inflammation can activate self-reactive CD8(+) T cells and induce autoimmunity. Here we show in a CD8(+) T cell-mediated model of type 1 diabetes that CD4(+)CD25(+) Treg cells prevent beta cell destruction following localized inflammation in the islets of Langerhans. These Treg cells accumulate preferentially in the pancreatic lymph nodes and islets but not other lymph nodes or spleen. PLN-derived Treg cells are extremely potent; only 2 x 10(3) cells are needed to prevent diabetes development, and their capacity to regulate is dependent on TNF-related activation induced cytokine-receptor activator of NFkappaB signals. Indeed, blockade of this pathway results in decreased frequency of CD4(+)CD25(+) Treg cells in the PLN, resulting in intra-islet differentiation of CD8(+) T cells into CTLs and rapid progression to diabetes. PMID- 11869681 TI - WIP deficiency reveals a differential role for WIP and the actin cytoskeleton in T and B cell activation. AB - WIP stabilizes actin filaments and is important for filopodium formation. To define the role of WIP in immunity, we generated WIP-deficient mice. WIP(minus sign/minus sign) mice have normal lymphocyte development, but their T cells fail to proliferate, secrete IL-2, increase their F-actin content, polarize and extend protrusions following T cell receptor ligation, and are deficient in conjugate formation with superantigen-presenting B cells and anti-CD3 bilayers. In contrast, WIP-deficient B lymphocytes have enhanced proliferation and CD69 expression following B cell receptor ligation and mount normal antibody responses to T-independent antigens. Both WIP-deficient T and B cells show a profound defect in their subcortical actin filament networks. These results suggest that WIP is important for immunologic synapse formation and T cell activation. PMID- 11869682 TI - Role for CCR7 ligands in the emigration of newly generated T lymphocytes from the neonatal thymus. AB - Most T lymphocytes are generated within the thymus. It is unclear, however, how newly generated T cells relocate out of the thymus to the circulation. The present study shows that a CC chemokine CCL19 attracts mature T cells out of the fetal thymus organ culture. Another CC chemokine CCL21, which shares CCR7 with CCL19 but has a unique C-terminal extension containing positively charged amino acids, failed to show involvement in thymic emigration. Neonatal appearance of circulating T cells was defective in CCL19-neutralized mice as well as in CCR7 deficient mice but not in CCL21-neutralized mice. In the thymus, CCL19 is predominantly localized in the medulla including endothelial venules. These results indicate a CCL19- and CCR7-dependent pathway of thymic emigration, which represents a major pathway of neonatal T cell export. PMID- 11869684 TI - Deltex1 redirects lymphoid progenitors to the B cell lineage by antagonizing Notch1. AB - Notch1 signaling drives T cell development at the expense of B cell development from a common precursor, an effect that is dependent on a C-terminal Notch1 transcriptional activation domain. The function of Deltex1, initially identified as a positive modulator of Notch function in a genetic screen in Drosophila, is poorly understood. We now demonstrate that, in contrast to Notch1, enforced expression of Deltex1 in hematopoietic progenitors results in B cell development at the expense of T cell development in fetal thymic organ culture and in vivo. Consistent with these effects, Deltex1 antagonizes Notch1 signaling in transcriptional reporter assays by inhibiting coactivator recruitment. These data suggest that a balance of inductive Notch1 signals and inhibitory signals mediated through Deltex1 and other modulators regulate T-B lineage commitment. PMID- 11869683 TI - Chronic intestinal inflammatory condition generates IL-10-producing regulatory B cell subset characterized by CD1d upregulation. AB - B cells possess a variety of immune functions that are involved in normal and abnormal immune responses, including autoimmune disorders. Through murine models of intestinal inflammation, we here demonstrate a B cell subset that is induced in gut-associated lymphoid tissues and is characterized by CD1d upregulation. This B cell subset appears under a chronic inflammatory environment, produces IL 10, and suppresses progression of intestinal inflammation by downregulating inflammatory cascades associated with IL-1 upregulation and STAT3 activation rather than by altering polarized T helper responses. This study indicates that B cells, by producing cytokines such as IL-10, can act as regulatory cells in immunologically mediated inflammatory reactions. PMID- 11869685 TI - A point mutation in the constant region of Ig lambda1 prevents normal B cell development due to defective BCR signaling. AB - Surface expression of B cell antigen receptors (BCRs) containing Ig and Igalpha/Igbeta generates signals required to transit discrete developmental checkpoints. The mechanism by which BCR components collaborate to initiate signals is still unclear. The expression of Iglambda1 in SJL mice is 50-fold lower than in other strains. Here, we demonstrate by gene targeting that a point mutation, which changes a glycine to a valine in the lambda1 constant region, is responsible for this defect. In vitro experiments show that Ig receptors bearing this mutation, while expressed normally, are deficient in signaling. These findings reveal a direct involvement of the Ig light chain (IgL) in B cell signaling and development beyond the requirement of light chains for BCR assembly. PMID- 11869686 TI - Dendritic cell development and survival require distinct NF-kappaB subunits. AB - Despite the established role of dendritic cells (DCs) in regulating T lymphocyte activation, intracellular mechanisms responsible for controlling DC function are largely undefined. Here, we have studied DCs from mice deficient in the p50, RelA, and cRel subunits of the immunomodulatory NF-kappaB transcription factor. Although DC development and function was normal in mice lacking individual NF kappaB subunits, development of doubly deficient p50(-/-)RelA(-/-) DCs was significantly impaired. In contrast, DCs from p50(-/-)cRel(-/-) mice developed normally, but CD40L- and TRANCE-induced survival and IL-12 production was abolished. Surprisingly, no significant impairment in MHC and costimulatory molecule expression was seen, despite significantly reduced kappaB site binding activity. These results therefore indicate essential, subunit-specific functions for NF-kappaB proteins in regulating DC development, survival, and cytokine production. PMID- 11869687 TI - A restricted subset of dendritic cells captures airborne antigens and remains able to activate specific T cells long after antigen exposure. AB - Mice sensitized for a Th2 response to Leishmania LACK antigen developed allergic airway inflammation upon exposure to LACK aerosol. Using multimers of I-A(d) molecules bound to a LACK peptide as probes, we tracked the migration of LACK specific Th2 cells to the airways. Elevated numbers of LACK-specific Th2 cells remained in the airways for 5 weeks after the last aerosol. Substantial numbers of DC presenting LACK peptides were found in the airways, but not in other compartments, for up to 8 weeks after antigen exposure. These LACK-presenting airway DC expressed CD11c and CD11b as well as high levels of surface molecules involved in uptake and costimulation. Taken together, our results may explain the chronic Th2 airway inflammation characteristic of allergic asthma. PMID- 11869688 TI - Constitutive expression of PU.1 in fetal hematopoietic progenitors blocks T cell development at the pro-T cell stage. AB - The essential hematopoietic transcription factor PU.1 is expressed in multipotent thymic precursors but downregulated during T lineage commitment. The significance of PU.1 downregulation was tested using retroviral vectors to force hematopoietic precursors to maintain PU.1 expression during differentiation in fetal thymic organ culture. PU.1 reduced thymocyte expansion and blocked development at the pro-T cell stage. PU.1-expressing cells could be rescued by switching to conditions permissive for macrophage development; thus, the inhibition depends on both lineage and developmental stage. An intact DNA binding domain was required for these effects. PU.1 expression can downregulate pre-Talpha, Rag-1, and Rag-2 in a dose-dependent manner, and higher PU.1 levels induce Mac-1 and Id-2. Thus, downregulation of PU.1 is specifically required for progression in the T cell lineage. PMID- 11869689 TI - PU.1 regulates expression of the interleukin-7 receptor in lymphoid progenitors. AB - Development of the lymphoid system is dependent on the Ets family transcription factor PU.1. We demonstrate that PU.1(-/-) hematopoietic progenitors fail to express IL-7Ralpha transcripts. Promoter and chromatin crosslinking analyses suggest that PU.1 directly regulates transcription of the IL-7Ralpha gene. Retroviral transduction of IL-7Ralpha into PU.1(-/-) progenitors restores IL-7 dependent proliferation and induces, at low frequency, the generation of pro-B cells undergoing an apparently normal program of differentiation. Although the related factor Spi-B can substitute for PU.1 in early B cell development, it is not required. These results demonstrate that PU.1 functions to regulate early B cell development in part by controlling the expression of the IL-7Ralpha gene. PMID- 11869691 TI - Pharmacoeconomics for surgeons. PMID- 11869690 TI - CD4(+)CD25(+) immunoregulatory T cells: gene expression analysis reveals a functional role for the glucocorticoid-induced TNF receptor. AB - CD4(+)CD25(+) immunoregulatory T cells represent a unique lineage of thymic derived cells that potently suppress both in vitro and in vivo effector T cell function. We analyzed CD4(+)CD25(+) and CD4(+)CD25(-) T cells by DNA microarray, identifying 29 genes differentially expressed in the resting subpopulations, and 77 that were differentially expressed following activation. Most of these genes were elevated in the CD4(+)CD25(+) population, suggesting a previously activated phenotype. Among these were a number of genes that antagonize signaling, including members of the SOCS family, which may contribute to their anergic phenotype. Multiple cell surface receptors also had increased expression in CD4(+)CD25(+) cells, including GITR, a member of the TNF receptor superfamily. Importantly, antibodies to GITR abrogated suppression, demonstrating a functional role for this receptor in regulating the CD4(+)CD25(+) T cell subset. PMID- 11869693 TI - Shouldn't cost be considered a significant issue in medical care? PMID- 11869692 TI - Treatment of Helicobacter pylori infection in patients with peptic ulcer disease. AB - Helicobacter pylori is an organism that is thought to be important in the pathophysiology of peptic ulcer disease and gastritis. Eradication of the organism is useful in clinical cure of infected patients. Efficacious regimens generally include an antisecretory agent combined with two to three antimicrobials. The main determinant of overall cost of treatment is the rate of eradication of the organism. Impediments to patient compliance include intensive dosage schedules and drug side effects. Resistance can occur to the commonly used antibiotics but can usually be overcome with an altered regimen. PMID- 11869695 TI - Surgical clips to help identify anatomy after a Nissen fundoplication. AB - Laparoscopic fundoplication has had a remarkable growth in the treatment of gastroesophageal reflux disease. But a failure rate of more than 5% has led to increasing numbers of patients with recurrent symptoms presenting for corrective surgery. A simple technique of placing surgical clips on the sutures used to place the wrap helps identify the anatomy during contrast studies and helps plan for corrective surgery. PMID- 11869694 TI - Clinical and functional results after elective colonic resection in 75 consecutive patients with diverticular disease. AB - BACKGROUND: Functional results after elective colonic resection in patients with diverticular disease have seldom been studied. METHODS: Seventy-five consecutive patients were reviewed and sent a questionnaire about abdominal symptoms and functional results. Possible associations between patients' characteristics and postoperative complications or functional outcome were analyzed. RESULTS: Major complications including anastomotic leakage, bleeding, and bowel obstruction occurred in 10 patients (13%). Six patients (8%) had recurrent diverticulitis. No significant associations were found between clinical characteristics and postoperative complications or recurrent disease. Fifty patients classified their final result as excellent or good. Functional symptoms or symptoms suggestive of irritable bowel syndrome before the operation predicted a less successful result (P <0.05). CONCLUSIONS: Elective surgery in patients with diverticular disease was hampered by postoperative complications but resulted in most cases in good functional outcome and a low rate of recurrent disease. Those with functional bowel symptoms before surgery had significantly worse results. PMID- 11869696 TI - Laparoscopic diagnosis and repair of asymptomatic bilateral inguinal hernias. AB - BACKGROUND: This study was designed to investigate age, sex, and side of hernia presentation at clinical examination as potential intrinsic risk factors for bilateral inguinal hernia (BIH), and to quantify the characteristics of clinical examination versus laparoscopy as a diagnostic tool for BIH. METHODS: A cross sectional study was utilized to analyze 99 consecutive patients undergoing laparoscopic inguinal hernia repair. RESULTS: The incidence of BIH based on clinical examination alone was 49%, compared with 71% laparoscopically. Clinical examination of BIH resulted in 69% sensitivity, 100% specificity, 100% predictive value for bilateral diagnosis, and 57% predictive value for unilateral diagnosis. Left hernia presentation at clinical examination (prevalence rate ratio = 10.5, 95% confidence interval: 3.6 to 30.7) and male sex (prevalence rate ratio = 6.6, 95% confidence interval: 1.3 to 35.0) were found to be independent risk factors for BIH. CONCLUSIONS: Laparoscopy yields detection of BIH that would be missed by clinical examination alone. Furthermore, left-sided hernia and male sex were associated with BIH. PMID- 11869697 TI - The incision of choice for pregnant women with appendicitis is through McBurney's point. AB - BACKGROUND: There is uncertainty over the optimal incision for gravid patients with appendicitis. METHODS: Data were collected retrospectively from January 1, 1995, through December 31, 2000, on 374 women of childbearing age who underwent appendectomies. Of these, 23 gravid patients were evaluated. RESULTS: Eighteen incisions were made over McBurney's point and five were created superior to McBurney's point. Patients in the third trimester of pregnancy all received incisions over McBurney's point. The appendix was located without difficulty in all 4 of the third trimester patients. The appendix was easily located in 94% of the incisions made through McBurney's point and 80% of the incisions made above McBurney's point. CONCLUSIONS: Our clinical experience indicates that the incision for the removal of the appendix in pregnant patients in all trimesters can be successfully made over McBurney's point. PMID- 11869698 TI - Sentinel lymph node biopsy results in less postoperative morbidity compared with axillary lymph node dissection for breast cancer. AB - BACKGROUND: This study was designed to compare the postoperative morbidity and socioeconomic impact of sentinel lymph node biopsy (SLNB) with axillary lymph node dissection (ALND) in patients with early stage breast cancer. METHODS: A prospective, nonrandomized, controlled study was designed to include patients who underwent breast conservation surgery and SLNB +/- ALND. Group A consisted of patients who had a negative SLNB and did not go on to completion ALND. Group B consisted of patients who underwent a SLNB followed by a completion ALND because either (1) their sentinel node contained cancer or (2) they were within the validation phase of our institution's sentinel lymph node protocol. Patients were evaluated with a questionnaire and underwent a standardized physical examination to determine arm circumference. RESULTS: Data were obtained from 96 patients with a mean follow-up period of 15 months (range 8 to 29). Significant differences were seen in subjective measurements of arm complaints and arm numbness (P <0.001), with fewer complaints reported in group A. The difference in mid-bicep and antecubital fossa circumferences was significant when comparing the ratio of the procedure arm with the nonprocedure arm and when subtracting the nonprocedure arm from the procedure arm (P <0.003 and P <0.016, respectively) in favor of group A. Axillary surgery was performed as an outpatient procedure in 88% of group A patients, compared with 15% in group B (P <0.001). Furthermore, 71% of group A patients returned to "normal activity" in less than 4 days, in comparison with 7% of group B (P <0.001). CONCLUSIONS: SLNB results in less postoperative morbidity in terms of subjective arm complaints and mid-arm swelling. Expeditious return to work or normal activity after SLNB has potentially significant socioeconomic consequences. PMID- 11869700 TI - Duodenum-preserving pancreatic head resection leads to relief of common bile duct stenosis. AB - BACKGROUND: Common bile duct stenosis (CBDS) is one of the most frequent complications in chronic pancreatitis with inflammatory mass in the head of the pancreas (IMH). METHODS: A total of 474 patients who underwent duodenum preserving pancreatic head resection (DPPHR) between 1982 and 1998 were reevaluated; 219 patients (46%) with a mean duration of the disease of 45 months had a radiologically proven CBDS. RESULTS: One patient (0.5%) died of septic complications in the early postoperative course, 15 patients (6.8%) had to be reoperated on for complications. A follow-up investigation of 143 patients (92%) revealed a late mortality of 12%; no patient died of biliary complications. Seventy-five percent of the patients were completely free of pain, and 85% of the patients had a constant or even increasing body weight. CONCLUSIONS: The high percentage of pain-free patients with improved physical status and economical rehabilitation demonstrates the improvement of the quality of life after DPPHR for complicated chronic pancreatitis. PMID- 11869699 TI - Deliberate hypoxic pelvic and limb chemoperfusion in the treatment of recurrent melanoma. AB - BACKGROUND: The treatment of patients with advanced or recurrent pelvic melanoma, which are often associated with lesions in the lower limbs, is still unsatisfactory and controversial. A simplified hypoxic pelvic and limb perfusion has been recently recommended to provide therapeutic options for palliation and possibly cure. METHODS: A nonrandomized and noncontrolled phase II experimental study was performed in 11 patients with symptomatic unresectable recurrent melanoma of the pelvis and limb. Patients were submitted to hypoxic pelvic and limb perfusion with 25 mg/m(2) of melphalan, 50 mg/m(2) of cisplatin, 300 mg/m(2) of dacarbazine, and 75 mg/m(2) of epirubicin by means of a simplified balloon occlusion technique. Response rate and time to disease progression were the primary endpoints; overall survival was the secondary endpoint. RESULTS: During the procedures there were no technical, hemodynamic, or vascular complications, and no deaths occurred during surgery or in the postoperative period. Response rate was 82% (95% confidence interval, 58% to 100%). Median time to disease progression was 12 months (range 9 to 30 months). Three-year overall survival was 34%. CONCLUSIONS: Hypoxic pelvic and limb perfusion is a safe and good palliative treatment for patients with unresectable recurrent melanoma. Further studies are necessary to to confirm these data and to establish if refinements can be made with acceptable toxicity. PMID- 11869701 TI - Prevention of pancreatic anastomotic leakage after pancreaticoduodenectomy. AB - BACKGROUND: Leakage at the pancreaticoenteric anastomosis remains a common and serious complication after pancreaticoduodenectomy. Over the past decade, various measures directed towards prevention of pancreatic leakage have been studied. This article reviews the available data on the efficacy of these measures. DATA SOURCES: The Medline database from 1990 to 2000 was searched for studies on the prevention of pancreatic anastomotic leakage, and the bibliographies of the articles were reviewed for additional references. RESULTS: A meta-analysis of the results of prophylactic octreotide in preventing pancreatic fistula after pancreaticoduodenectomy from data available in three randomized controlled studies yielded an odds ratio of 1.08 (95% confidence interval 0.64 to 1.84). Pending further trials to clarify its role, the routine use of octreotide in pancreaticoduodenectomy cannot be recommended. Retrospective or nonrandomized prospective studies suggested that technical modifications such as duct-to-mucosa anastomosis, pancreaticogastrostomy and external pancreatic duct stenting may reduce the leakage rate, but there is a paucity of randomized trials. A randomized trial comparing pancreaticogastrostomy and pancreaticojejunostomy did not reveal a significant difference in the leakage rate. CONCLUSIONS: Further randomized controlled studies are required to determine the optimum technique of pancreaticoenteric anastomosis after pancreaticoduodenectomy. PMID- 11869702 TI - Ten-year outcome of patients with very small abdominal aortic aneurysm. AB - BACKGROUND: The long-term fate of very small abdominal aortic aneurysms (AAA) is not well known. METHODS: Forty-one patients with asymptomatic small AAA (range 25 to 40 mm) underwent ultrasonographic surveillance. RESULTS: The median follow-up period was 7.3 years. The median linear aneurysm expansion rate was 2.0 mm/year (range 0 to 8.4). Three patients experienced aneurysm rupture (7.3%) which resulted in 1 patient'death. Thirteen patients underwent aneurysm repair (31.7%) and 1 patient died postoperatively (7.7%). The survival rate at 10-year follow-up was 59.0%. The survival rate free from aneurysm rupture and repair at 10-year follow-up was 69.9%. The median time for occurrence of aneurysm rupture was 4.9 years (range 1.8 to 10.5) and the need for aneurysm repair was 4.5 years (range 1.4 to 10.4). CONCLUSIONS: The fate of very small AAA is to slowly enlarge in size, sometimes threatening the patient's life. These observations underline the importance of continuous surveillance and the potential benefits of any medical treatment in this patient population. PMID- 11869703 TI - Gallstone ileus with cholecystoduodenal fistula. PMID- 11869704 TI - Liver resection with repair of major hepatic veins. AB - BACKGROUND: Liver resections for tumors adjacent to major hepatic veins often require reconstruction of venous wall defects. We describe a new operative approach that facilitates repair of major hepatic veins during hepatectomies. METHODS: In 3 cases of liver tumors, the resection line had to include partially the wall of the right hepatic vein, middle hepatic vein and left hepatic vein of the preserved liver. The procedure was carried out by employing portal triad clamping combined with extrahepatic occlusion of the hepatic veins. Venous grafts for vascular repair were harvested from the inferior mesenteric vein. RESULTS: In all 3 patients, histology showed tumor-free resection margins. Follow-up of 32 to 42 months revealed no recurrence and excellent liver function. CONCLUSIONS: Combination of selective hepatic vascular exclusion with venous repair techniques, facilitates extensive liver resections in patients with tumors adjacent to the major hepatic veins and maximizes preservation of healthy liver tissue. PMID- 11869705 TI - Percutaneous transhepatic cholecystostomy and delayed laparoscopic cholecystectomy in critically ill patients with acute calculus cholecystitis. AB - BACKGROUND: The ultimate therapy for acute cholecystitis is cholecystectomy. However, in critically ill elderly patients the mortality of emergency cholecystectomy may reach up to 30%. Open cholecystostomy performed under local anesthesia was considered to be the procedure of choice for treatment of acute cholecystitis in high-risk patients. In recent years, ultrasound- or computed tomography (CT)-guided percutaneous transhepatic cholecystostomy (PTHC) replaced open cholecystostomy for the treatment of acute cholecystitis in critically ill patients. METHODS: The aim of the present study was to evaluate the results of a 5-year protocol using PTHC followed by delayed laparoscopic cholecystectomy for the treatment of acute cholecystitis in critically ill patients. We reviewed the charts of 55 patients who underwent PTHC at the Hadassah University Hospital Mount Scopus during the years 1994 to 1999. RESULTS: The main indications for PTHC among this group of severely sick and high-risk patients was biliary sepsis and septic shock in 23 patients (42%); and severe comorbidities in 32 patients (58%). The median age was 74 (32 to 98) years, 33 were female and 22 male. Successful biliary drainage by PTHC was achieved in 54 of 55 (98%) of the patients. The majority of the patients (31 of 55) were drained transhepaticlly under CT guidance. The rest, (24 of 55) were drained using ultrasound guidance followed by cholecystography for verification. Complications included hepatic bleeding that required surgical intervention in 1 patient and dislodgment of the catheter in 9 patients that was reinserted in 2 patients. Three patients died of multisystem organ failure 12 to 50 days following the procedure. The remaining 52 patients recovered well with a mean hospital stay of 15.5 plus minus 11.4 days. Thirty-one patients were able to undergo delayed surgery: 28 underwent laparoscopic cholecystectomy of whom 4 (14%) were converted to open cholecystectomy. This was compared with a 1.9% conversion rate in 1,498 elective laparoscopic cholecystectomies performed at the same time period (P = 0.012). Another 3 patients underwent planned open cholecystectomy, 1 urgent and 2 combined with other abdominal procedures. There was no surgery associated mortality, severe morbidity, or bile duct injury. CONCLUSIONS: The use of PTHC in critically ill patients with acute cholecystitis is both safe and effective. PMID- 11869706 TI - Peritoneal fibrinolytic activity in peritonitis. AB - BACKGROUND: Peritonitis may cause a reduction in abdominal fibrinolytic activity. The reduced local fibrinolysis seems to be an important process in the subsequent development of adhesion formation. The aim of the study was to evaluate peritoneal fibrinolytic capacity in inflamed and normal peritoneum. METHODS: Peritoneal biopsy specimens were taken at the beginning of operation from 15 patients with peritonitis and 10 patients who underwent elective operation. Levels of tissue-type plasminogen activator (tPA), urokinase-type plasminogen activator (uPA), plasminogen activator inhibitor (PAI) type 1 (PAI-1) and type 2 (PAI-2), and tPA/PAI complex in tissue extracts were determinated by commercially available enzyme-linked immunosorbent assay kits. RESULTS: tPA was significantly reduced in peritonitis compared with normal peritoneum (P <0.001), whereas it was found that the levels of PAI-1, PAI-2, uPA, and tPA/PAI complex in peritonitis were significantly higher than those in normal controls. CONCLUSIONS: Plasminogen activator activity was significantly reduced in peritoneal biopsy samples from patients with peritonitis compared with those from patients without peritonitis. PMID- 11869708 TI - Intrapelvic complications after total hip arthroplasty failure. AB - BACKGROUND: Severe total hip arthroplasty failure with central migration of prosthetic components is uncommon. If perforation of the medial acetabular wall occurs, injuries of intrapelvic structures may result. DATA SOURCES: A meta analysis of the English literature was performed. A human pelvic cadaver was used to demonstrate the proximity of intrapelvic structures to a centrally dislocated cup. RESULTS: Fifty cases of intrapelvic injury were identified. Structures involved most frequently were the external iliac artery and the bladder. The most common types of complication included fistula formation, development of a false aneurysm, and hemorrhage. The human cadaver pelvis demonstrated the proximity of intrapelvic vessels, the bladder, the ureter, the vagina, the deferent duct, the sigmoid colon, the rectum, and the sciatic nerve to an intrapelvically intruded prosthesis. CONCLUSIONS: Failed total hip replacements should be considered to cause damage to pelvic viscera. PMID- 11869707 TI - Role of integrins and intracellular adhesion molecule-1 in lung injury after intestinal ischemia-reperfusion. AB - BACKGROUND: We tested the hypothesis that lung injury after intestinal ischemia reperfusion (IR) requires the activation of CD11/CD18 glycoprotein complex and its ligand, intracellular adhesion molecule-1 (ICAM-1), on pulmonary endothelial surface. METHODS: Rats were assigned to one of six groups including sham operation, intestinal IR (60/120 min) and IR plus treatment with one of the following monoclonal antibodies against CD11a, CD11b, CD18, and ICAM-1. Pulmonary microvascular permeability, neutrophil accumulation, and expression of adhesion molecules were evaluated. RESULTS: Intestinal IR resulted in lung injury characterized by a marked increase in microvascular permeability, neutrophil accumulation and upregulated expression of leukocyte integrins and ICAM-1. The increase in pulmonary microvascular permability and neutrophil accumulation elicited by intestinal reperfusion was effectively prevented by administration of blocking antibodies against ICAM-1, CD11, and CD18. CONCLUSIONS: These results indicate that adhesion molecules contribute to the lung injury after intestinal IR. Immunoneutralization of certain of these adhesion molecules may prevent intestinal IR-induced lung injury. PMID- 11869709 TI - The clinicopathological significance of histologic vascular invasion in differentiated thyroid carcinoma. AB - BACKGROUND: Clinicopathological correlation and clinical importance of histologic vascular invasion in differentiated thyroid carcinoma have not been fully examined. METHODS: Histologic vascular invasion of 256 differentiated thyroid carcinomas was examined using Victoria-blue hematoxylin-eosin staining. RESULTS: Vascular invasion was found in 120 patients and was independently related to lymph node metastasis (P <0.0001), extrathyroidal invasion (P = 0.0003) and differentiation (P = 0.0183). Patients with vascular invasion more frequently relapsed than those without (P = 0.0069). The disease-free survival of patients with vascular invasion (15.6 +/- 1.8 years) was shorter than that of patients without vascular invasion (20.5 +/- 0.9 years, P = 0.0001). In multivariate analysis, vascular invasion is an independent prognostic factor for disease-free survival, but not for overall survival. CONCLUSIONS: These data suggest that histologic vascular invasion is associated with clinicopathologically aggressive thyroid carcinomas with lymphatic and hematogenous spread and is a prognostic factor for disease-free survival. PMID- 11869711 TI - A 15-year retrospective study of hepatic resection for stage IV-A hepatocellular carcinoma shows value in hepatitis B negative patients. AB - BACKGROUND: The aim of this study is to identify the risk factors of survival and recurrence after curative hepatic resection for stage IV-A hepatocellular carcinoma (HCC). METHODS: Sixty-five patients with stage IV-A HCC who underwent curative hepatic resection and discharged from hospital were enrolled in this retrospective study. Prognostic factors were evaluated by univariate and multivariate analysis. Clinicopathologic features and survival with stage IV-A HCC were compared with those of 290 patients with stage I to III HCC who underwent curative hepatic resection during the same period. RESULTS: Disease free and overall survival for patients with stage IV-A HCC was significantly lower than for those with stage I to III HCC. Positive hepatitis B virus (HBV) surface antigen was an independent prognostic factor of poor disease-free and overall survivals in patients with stage IV-A HCC. There were no significant differences in the disease-free and overall survivals between non-HBV-related stage IV-A HCC and stage I to III HCC. CONCLUSIONS: Even for patients with highly advanced HCC, curative hepatic resection may be a feasible therapeutic option for those with non-HBV-related HCC. PMID- 11869710 TI - Three-dimensional computer images of stomach diseases. PMID- 11869713 TI - Acute ovarian torsion in children. PMID- 11869715 TI - Whole gut washout ameliorates progression of acute experimental pancreatitis. PMID- 11869716 TI - Clinical role of noncontrast helical computed tomography in diagnosis of acute appendicitis. PMID- 11869719 TI - Results and complications after adjustable gastric banding. PMID- 11869721 TI - The scope of linguistic generalizations: evidence from Hebrew word formation. AB - Does the productive use of language stem from the manipulation of mental variables (e.g. "noun", "any consonant")? If linguistic constraints appeal to variables, rather than instances (e.g. "dog", "m"), then they should generalize to any representable novel instance, including instances that fall beyond the phonological space of a language. We test this prediction by investigating a constraint on the structure of Hebrew roots. Hebrew frequently exhibits geminates (e.g. ss) in its roots, but it strictly constraints their location: geminates are frequent at the end of the root (e.g. mss), but rare at its beginning (e.g. ssm). Symbolic accounts capture the ban on root-initial geminates as *XXY, where X and Y are variables that stand for any two distinct consonants. If the constraint on root structure appeals to the identity of abstract variables, then speakers should be able to extend it to root geminates with foreign phonemes, including phonemes with foreign feature values. We present findings from three experiments supporting this prediction. These results suggest that a complete account of linguistic processing must incorporate mechanisms for generalization outside the representational space of trained items. Mentally-represented variables would allow speakers to make such generalizations. PMID- 11869722 TI - Conceiving of entities as objects and as stuff. AB - What is the difference between conceiving of an entity as an object of some kind and conceiving of it as an amount of solid stuff? We propose that the difference lies in how one thinks about the entity's structure. Object construals require thinking of the structure as being nonarbitrary, whereas substance construals require thinking of the structure as being arbitrary. We report six experiments that provide empirical support for this proposal. Regularity of structure, repetition of structure, and the existence of structure-dependent functions, all of which provide reasons to consider the structure of an entity to be nonarbitrary, were shown to bias participants towards object construals. We also discuss how the proposed account of what it means to construe an entity as an object or as some stuff can account for a range of findings in the literature on lexical development. These include the relation between cohesiveness and ontological category, shape and ontological category, and complexity of shape and ontological category. Finally, we discuss the nature of construals and the relation of object and substance construals to the physical, design, and intentional stances. PMID- 11869723 TI - Bootstrapping the lexicon: a computational model of infant speech segmentation. AB - Prelinguistic infants must find a way to isolate meaningful chunks from the continuous streams of speech that they hear. BootLex, a new model which uses distributional cues to build a lexicon, demonstrates how much can be accomplished using this single source of information. This conceptually simple probabilistic algorithm achieves significant segmentation results on various kinds of language corpora - English, Japanese, and Spanish; child- and adult-directed speech, and written texts; and several variations in coding structure - and reveals which statistical characteristics of the input have an influence on segmentation performance. BootLex is then compared, quantitatively and qualitatively, with three other groups of computational models of the same infant segmentation process, paying particular attention to functional characteristics of the models and their similarity to human cognition. Commonalities and contrasts among the models are discussed, as well as their implications both for theories of the cognitive problem of segmentation itself, and for the general enterprise of computational cognitive modeling. PMID- 11869724 TI - A tale of two theories: response to Fisher. PMID- 11869725 TI - Why are we good at detecting cheaters? A reply to Fodor. AB - It is argued that the recent criticism by Fodor (Cognition 75 (2000) 29) of "cheater detection" in the Wason selection task is based upon a false presumption about what the task entails. Fodor compares two different ways of presenting the task, rather than two different task domains (social and non-social). Consequently, the conclusion that the selection task can tell us nothing about either the architecture or the history of cognition is invalid. Fodor's explanation of the Wason selection task is examined experimentally and compared to predictions derived from social contract theory (Cognition 31 (1989) 187). It is concluded that, although Fodor's variant of the Wason selection task improves performance, this improvement is independent of the task domain and is insufficient to account for the "cheater detection" effect. PMID- 11869727 TI - Tool-use changes multimodal spatial interactions between vision and touch in normal humans. AB - In a visual-tactile interference paradigm, subjects judged whether tactile vibrations arose on a finger or thumb (upper vs. lower locations), while ignoring distant visual distractor lights that also appeared in upper or lower locations. Incongruent visual distractors (e.g. a lower light combined with upper touch) disrupt such tactile judgements, particularly when appearing near the tactile stimulus (e.g. on the same side of space as the stimulated hand). Here we show that actively wielding tools can change this pattern of crossmodal interference. When such tools were held in crossed positions (connecting the left hand to the right visual field, and vice-versa), the spatial constraints on crossmodal interference reversed, so that visual distractors in the other visual field now disrupted tactile judgements most for a particular hand. This phenomenon depended on active tool-use, developing with increased experience in using the tool. We relate these results to recent physiological and neuropsychological findings. PMID- 11869728 TI - Visual statistical learning in infancy: evidence for a domain general learning mechanism. AB - The rapidity with which infants come to understand language and events in their surroundings has prompted speculation concerning innate knowledge structures that guide language acquisition and object knowledge. Recently, however, evidence has emerged that by 8 months, infants can extract statistical patterns in auditory input that are based on transitional probabilities defining the sequencing of the input's components (Science 274 (1996) 1926). This finding suggests powerful learning mechanisms that are functional in infancy, and raises questions about the domain generality of such mechanisms. We habituated 2-, 5-, and 8-month-old infants to sequences of discrete visual stimuli whose ordering followed a statistically predictable pattern. The infants subsequently viewed the familiar pattern alternating with a novel sequence of identical stimulus components, and exhibited significantly greater interest in the novel sequence at all ages. These results provide support for the likelihood of domain general statistical learning in infancy, and imply that mechanisms designed to detect structure inherent in the environment may play an important role in cognitive development. PMID- 11869730 TI - The social implications of an ageing population. Introduction. AB - The growing proportion of older people, especially the very old, is often thought of in negative terms such as the need for more health care. But the implications are wider and affect the future of family care as well as issues of intergenerational relations, employment, living arrangements and attitudes. This paper looks at the social implications of an ageing population on people and on services. PMID- 11869731 TI - Evolution of ageing. AB - Explaining why ageing occurs is a solution to the longstanding enigma of the role of senescence in nature. Even after half a century of progress, this solution continues to unfold. Evolution theory argues strongly against programmed ageing, suggesting instead that organisms are programmed for survival, not death. In the current view, ageing results from the twin principles that (i) the force of natural selection declines with age, and (ii) longevity requires investments in somatic maintenance and repair that must compete against investments in growth, reproduction and activities that might enhance fitness. In addition to explaining why ageing occurs, the evolutionary theory also provides insight into the mechanisms underlying the complex cellular and molecular changes that contribute to senescence, as well as an array of testable predictions. Some of the most interesting current problems are to understand how the genetic factors influencing ageing and longevity are predicted to respond to fluctuating environments, such as temporary periods of famine, as well as to other kinds of spatial and/or temporal heterogeneity. Rapid progress in human genomics raises the prospect of greatly increasing our knowledge of the determinants of human longevity. To make progress in understanding the role and evolution of genetic and non-genetic factors in human longevity, we need more detailed theoretical studies of how intra-population variables, such as socio-economic status, influence the selection forces that shape the life history. PMID- 11869732 TI - Ageing in plants. AB - Ageing in green plants differs in some fundamental ways from the process in animals. The seasonal cycle and persistence of a plant is governed by a combination of the determinate or indeterminate status of meristems (growth centres) and the cell death and disposal strategies employed by plants to generate well-adapted anatomies and morphologies. The degree of perenniality depends on the balance between exploratory growth and the wave of tissue death that succeeds it, and extremes of longevity can arise by relatively minor changes in the quantitative relationship between growth and death. The senescence and elimination of organs and tissues are related to the internal reallocation of resources but are programmed phases in the integrated development of the whole plant and do not represent a kind of ageing by stress or starvation. Meristems of long-lived plants accumulate genetic damage but selection mechanisms exist within the organism to control genetic load, and even to exploit somatic mutations that confer adaptive benefits. It is concluded that most plants do not age in the strict gerontological sense and that extremely long-lived forms like trees and clonal creeping perennials are sustained by selection and correction at the level of semi-autonomous cell lineages. PMID- 11869733 TI - Aging in fungi: role of mitochondria in Podospora anserina. AB - In experimental gerontology, there is a long tradition in the use of both unicellular and filamentous species of fungi. In the last three decades, biochemical, genetic and molecular approaches have proved very fruitful in elucidating different aspects of ageing. It was shown that various genes and molecular pathways are involved in life span control. The oxygenic energy metabolism plays a central role. During mitochondrial energy transduction, reactive oxygen species (ROS) are generated as by-products. These molecules are able to damage all cellular compounds leading to cellular dysfunctions. Within certain limits, however, cells are able to cope with ROS-related problems. First, ROS scavengers can be induced which are effective in lowering the molecular burden of ROS on cellular functions. Second, if damage occurs, specific repair mechanisms and the general turnover of affected molecules can maintain cellular functions. Finally, if damage of essential components is too severe, cells may induce specific pathways to compensate for the corresponding impairments. A coordinated interaction between different cellular compartments is involved in these processes. In this review I shall concentrate on the ageing in the filamentous ascomycete Podospora anserina. It is clear that both environmental as well as genetic traits are involved in the control of life span and that mitochondrial-nuclear interactions play a paramount role. PMID- 11869734 TI - Stress resistance as a determinate of C. elegans lifespan. AB - It is difficult to exaggerate the progress that has been made in biogerontology over the last 15 years. As with all scientific revolutions, a few experiments in a small number of laboratories have changed the way in which we think about and design experiments. As a result of these experiments, there is much evidence to suggest that a rudimentary understanding of some of the processes that cause aging will be available in the next decade. One particular area of progress is the molecular genetics of lifespan. Although one may draw some distinctions between chronological lifespan and normal aging, extended lifespan remains one of the best indicators that an intervention in an aging process has been made. The isolation of a long-lived variant of a laboratory invertebrate is now essentially a trivial project but the information obtained from this approach is proving invaluable. As with most other biological problems, the most important experimental developments are coming from studying simple organisms in a reductionist fashion. PMID- 11869735 TI - The evolution of senescence in fish. AB - Fish are over-represented among the vertebrates that are known to live over 100 years. Such trends in maximum longevity suggest that fish also experience delayed senescence relative to other vertebrates. Most applications of evolutionary theory suggest that the extrinsic mortality rate, or mortality that is attributable to external factors like disease or predation, is the major factor that shapes the evolution of senescence, so that the only way to evolve delayed senescence is to experience lower rates of extrinsic mortality. We propose instead that fish are more inclined to evolve delayed senescence because they have indeterminate growth and, as a consequence, have the capacity for a substantial increase in fecundity with age. It is thus the combined effects of the expectation of survival and fecundity that shape the evolution of senescence, as originally proposed by Williams (1957). We also argue that fish share many of the same general mechanisms that shape the evolution of senescence in other vertebrates. We support this argument with statistical analyses of life history data that show the same patterns of correlation among traits as seen in birds and mammals. PMID- 11869736 TI - Genes, telomeres and mammalian ageing. AB - Although there appear to be several influences, which contribute to the ageing of mammals, the role of DNA appears to be pivotal. There is increasing evidence that oxidative damage is an important factor in producing mutations in genes, shortening telomeres and damaging mitochondrial DNA. Accumulation of mutations in genomic DNA could result in the gradual decline in cellular function, which is exhibited in a variety of tissues. The random nature of these mutations, could also offer an explanation for differences in the degree and time of onset of age related changes, exhibited by different individuals. Shortening of telomeres, caused by oxidative damage or the end-replication problem, could result in the accumulation of post-mitotic cells in-vivo during ageing. This might impair certain aspects of physiology, such as wound healing. Mutation of mitochondrial DNA may also be important in causing loss of cells in post-mitotic tissues such as muscle or brain. In addition changes in the redox state during the life of an animal may alter transcription factor activities, leading to consistent changes in the gene expression profiles of mammalian tissues. The latter could explain consistent age-related changes that have been observed in cell structure and physiology. Although all of these mechanisms may make a contribution to ageing, it is likely that it is the interplay between them that produces the most prominent effects. PMID- 11869737 TI - Molecular mechanisms of skin ageing. AB - Cutaneous ageing is a complex biological phenomenon consisting of two components; intrinsic ageing, which is largely genetically determined and extrinsic ageing caused by environmental exposure, primarily UV light. In sun-exposed areas, these two processes are superimposed. The process of intrinsic skin ageing resembles that seen in most internal organs and is thought to involve decreased proliferative capacity leading to cellular senescence, and altered biosynthetic activity of skin derived cells. Extrinsic ageing, more commonly termed photoageing, also involves changes in cellular biosynthetic activity but leads to gross disorganisation of the dermal matrix. The molecular mechanisms underlying some of these changes are now beginning to be unravelled and are discussed. As these mechanisms are identified, further insights into the underlying processes of skin ageing should emerge and better strategies to prevent the undesirable effects of age on skin appearance should follow. PMID- 11869738 TI - Ageing of the brain. AB - The brains of individuals who are cognitively normal show age-related changes that include an overall reduction in the brain volume and weight and enlargement of the brain ventricles. These changes are partly the result of nerve cell loss but accurate estimates of neuronal loss are notoriously difficult to make. There is loss of synapses and dendritic pruning in the aged brain but in selected areas rather than globally. Neurofibrillary tangles and senile plaques are the neuropathological hallmark of Alzheimer's disease in which they are more abundant and widespread than in the brains of intellectually intact elderly people. Alzheimer's disease has, therefore, been regarded as accelerated brain ageing, however, since there is a strong genetic contribution to developing the disease it implies that it may not be the inevitable, even if frequent, consequence of old age. The interplay between genetic and environmental factors probably determines the degree of pathological brain ageing and whether or not individuals develop dementia. PMID- 11869739 TI - The changing nature of consultation-liaison in primary care: bridging the gap between research and practice. AB - Consultation-liaison (C-L) psychiatry is hypothesized to be a model of interface between primary care and specialist mental health services with significant advantages over other models of organizing mental health care. However, there are significant complexities in the definition and evaluation of this model. As well as discussing the definition of C-L in primary care, this paper highlights the gap between models of traditional C-L that are popular in practice and the increasingly complex models (based on chronic disease management) evaluated in research studies. It is hypothesized that traditional C-L approaches and newer models use different mechanisms of change to achieve their goals. The former focus on the relationships between primary care and specialist professionals, while the latter highlight the importance of the development of effective systems of delivering care. Although the latter may be crucial in enhancing the "efficacy" and "effectiveness" of these models in terms of clinician behavior change and patient outcome, the former may be crucial in terms of "dissemination" and "implementation" of these models from research contexts to routine care settings. PMID- 11869740 TI - Case review vs. usual care in primary care patients with depression: a pilot study. AB - This paper reports on the results of a pilot study comparing the efficacy of an experimental case review program between psychiatrists and primary care physicians (PCPs) with PCPs' usual care on the outcome of depressive disorders at 1-year. The secondary aim of the paper is to identify correlates of remission of depressive disorders at the 1-year follow-up. The experimental case review program consisted of 12 biweekly meetings of primary care physicians with a psychiatrist. Meetings lasted two hours and were based on review of cases identified at the interview as suffering from full-blown or subthreshold conditions. Subjects were assessed at baseline, 3 months and 1 year. The baseline assessment included the Composite International Diagnostic Interview, the Hamilton Depression (HDRS) and Anxiety (HARS) Rating Scales, the Medical Outcomes Study SF-36 and some forms to collect life events, major difficulties, social support, and ongoing treatments. At 3 months patients received by mail the General Health Questionnaire (GHQ-12) and at one year the baseline clinical assessment was replicated. Assessments were conducted on eighty cases with major, minor or subsyndromal depression (SSD). The experimental case review program was similar in efficacy to usual care, with an average reduction of 5.5 points on the HAMD. Significant reduction in the severity of depression was associated with a limited number of baseline characteristics, such as being married and having minor depression as compared to major depression and SSD. On the contrary, chronic physical illness at baseline predicted a lower reduction of HAMD scores at one year. While major difficulties and life events during the one-year interval between the two assessments significantly predicted a poorer outcome, social support at home was related to a better outcome. This pilot study indicates that a case review intervention program is not efficacious in a mixed sample of patients with subsyndromal to major depression representative of the current practice of primary case physicians. Still, it might be worthwhile for "difficult" patients selected by the PCPs. In our sample, psychosocial factors seem to be important predictors of outcome. Future trials and large naturalistic studies are needed to address this key point. PMID- 11869741 TI - The prevalence of nausea in the community: psychological, social and somatic factors. AB - Nausea is a commonly reported symptom with a point prevalence of about 12% in the community. Nausea is a prominent symptom in functional gastrointestinal disorders and patients with anxiety and depression frequently present gastrointestinal symptoms such as nausea and abdominal discomfort as their main problem when they consult a doctor. Functional gastrointestinal disorders are strongly related to anxiety and depressive disorders with a lifetime prevalence of 80--90% in samples from clinics of gastroenterology. This study examines the relationship between anxiety disorders, depressions and nausea in a large community sample. A questionnaire on physical and mental health and demographic and life-style factors was sent to all adults 20 years and above in Nord Trondelag county in Norway. A total of 94,197 questionnaires were sent, with 62,651 persons returning the questionnaire, a response rate of 66.5%. The presence of nausea, heartburn, diarrhea and constipation during the last year was recorded. Anxiety disorders and depressions were based on self-rating of the Hospital Anxiety and Depression Scale (HADS). Forty-eight per cent reported one or several gastrointestinal complaints during the last year. 12.5% complained of nausea. 15.3% had an anxiety disorder and 10.4% a depression based on HADS ratings. Presence of anxiety disorders carried the highest risk for nausea (OR 3.42). Presence of depression also increased the risk, but less than anxiety disorders (OR 1.47). Demographic factors, life-style factors and extra-gastrointestinal conditions did not reduce the OR of anxiety disorders and depressions to any significant extent. We found that the presence of anxiety disorders was the strongest risk factor for nausea. Depression also carried a certain risk while demographic factors, life-style factors and other somatic conditions did not carry any substantial risk for nausea. Differential diagnoses of nausea should therefore include both anxiety and depressive conditions. PMID- 11869743 TI - Active engagement in the consultation process. AB - This paper presents a case for the active, dynamic engagement of patients during consultations. Observations and hypotheses about the patient's predominant conflicts are used to formulate interventions early in the consultation process that are designed to convey meanings that make the patient's experience comprehensible to him. This includes attention to his personality and his world view. The activity of the consultant in appropriately communicating aspects of these formulations to the patient is designed to facilitate the process of self revelation and to permit a co-construction with the patient of a perspective on aspects of his life experience and his current problems. Active, dynamic engagement leads to the development of a bond between consultant and patient that facilitates the implementation of a treatment plan. Numerous case descriptions are included to illustrate this active approach as applied in different contexts. PMID- 11869742 TI - Physical recovery in anorexia nervosa: is this the sole purpose of a child and adolescent medical-psychiatric unit? AB - Patients with anorexia nervosa are discharged from inpatient medical psychiatric wards on achievement of their target weight ("physical recovery") or even earlier. However, recent studies have consistently revealed that a very high percentage relapse, usually within a year. We hypothesized that the more rapid pace of physical compared with psychological recovery in anorexia nervosa creates a gap or interim stage when inpatients are particularly vulnerable. This phase has its own identifiable characteristics and prognostic value. To counter this problem, we formulated a treatment model where patients in the "physical recovery" stage participate in a follow-up program within the inpatient unit itself concomitant with individual psychotherapy at the outpatient community service. The program helps to preserve the strong therapeutic alliance established during hospitalization and to maintain the patient's base of security against the loss of the "anorectic solution." The program provides a full range of services--inpatient, day-treatment and follow-up, adapting itself to the changing needs of the patient. It has been in effect in our unit for three years, and the rate of relapse has decreased dramatically. Further controlled studies are needed to confirm these findings. PMID- 11869744 TI - Assessment of a pilot course on the management of somatization disorder for family doctors. AB - Somatization disorder (SD) patients are difficult to treat and produce negative feelings in health professionals. Smith et al.'s guidelines have demonstrated cost-effectiveness in the treatment of these patients, but family doctors consider it difficult to put these into practice in the long term. The objective of this paper is to design and assess a pilot course, based on Smith's norms, to train general practitioners for the everyday management of SD patients in primary care. We have designed a 20-h practical course, using role-playing and video recording with standardized patients, and focusing on micro-skills recommended by the literature on the subject. Assessment of the efficacy of the course is made by evaluation of baseline and post course video recordings by researchers unaware of the order of the interviews. The comparison of baseline and post course assessments demonstrated a significant improvement in several key skills (giving a name to the illness, explaining the psychological and biological basis of the disease, and emphasizing stress reduction) but no change on others (explaining that SD is a well-known disorder, empowering the patient, not blaming the patient for his or her illness, and instilling hope). Finally, other skills such as assessing the patient's opinion of the illness, recognizing the reality of symptoms and informing that there is no life risk, were correctly done from the beginning and, therefore, showed no change. We found that training may facilitate the development of certain skills. However, some doctors' abilities might also require the use of techniques such as Balint groups to modify negative emotions, such as anger and fear, toward these patients. PMID- 11869745 TI - Mood disorder with mixed features due to vitamin B(12) and folate deficiency. AB - Vitamin B(12) and folate deficiency is often associated with affective disorders mainly of the depressive type. We report a case of a 42-year-old woman with a mood disorder with mixed depressed/manic features that was due to vitamin B(12) and folate deficiency. The psychopathology developed over a five-year period without hematologic or other overt clinical characteristics of pernicious anemia. Replacement treatment with vitamin B(12) and folate was rapidly followed by full clinical remission, electroencephalographic normalization and neuropsychological improvement. At a one-year follow-up this condition was stable. Consequently, patients who respond poorly to psychopharmacologic treatment and/or present with atypical mood symptoms would warrant determination of vitamin B(12) and folate serum levels. PMID- 11869746 TI - Lack of clinical relevance of routine chest radiography in acute psychiatric admissions. AB - To make clinically relevant recommendations for chest X-ray testing in acute psychiatric admissions, this study examined the current practice of this screening test in patients admitted to a University Hospital. The records of the 332 first consecutive admissions to the psychiatric ward were assessed. In 200 patients (60%) a chest X-ray was requested. The X-ray film was normal in 81.5% of patients. The remaining subjects presented abnormalities: nonrelevant in twenty seven (13.5% of the total), and relevant in eleven (5.5%). Since all these relevant abnormalities were already known, in no cases was the test followed by changes in therapy or by additional diagnostic procedures. In almost all cases this screening test was of no practical value. Our findings challenge the systematic indication of chest X-ray in acute psychiatric patients, and suggest that the number of tests performed and the cost of medical care could be reduced by a more efficient use of past medical history and physical examination criteria, without compromising the quality of patient care. PMID- 11869747 TI - Segmental dystonia as the sole manifestation of carbamazepine toxicity. PMID- 11869749 TI - Bipolarity from ancient to modern times: conception, birth and rebirth. AB - We review the history of bipolar disorders from the classical Greek period to DSM IV. Perhaps the first person who described mania and melancholia as two different phenomenological states of one and the same disease was the Greek physician of the 1st century AD, Aretaeus of Cappadocia. The modern concept of bipolar disorders was born in France, with the publications of and. Emil Kraepelin, however, in 1899, unified all types of affective disorders in 'manic-depressive insanity'; in spite of some opposition, Kraepelin's unitary concept was adopted worldwide. In the 1960s, however, the rebirth of bipolar disorders took place through the publications of Jules Angst, Carlo Perris, and George Winokur, who independently showed that there exist clinical, familial and course characteristics validating the distinction between unipolar and bipolar disorders; in addition, they verified several of the corresponding opinions of the Wernicke-Kleist-Leonhard school. The concept of unipolar and bipolar disorders has further advanced in the last three decades: landmark developments include the renaissance of Kraepelin's mixed states and of Kahlbaum's and Hecker's cyclothymia and related affective temperaments, the concept of soft bipolar spectrum (Akiskal), and the distinction of schizoaffective disorders into unipolar and bipolar forms. PMID- 11869750 TI - Lithium treatment at 52. AB - The paper reviews briefly and comments in detail on selected events in the history of lithium treatment. The events include Cade's discovery of lithium's antimanic action, the discovery and establishment of its prophylactic action, the ensuing debate, amelioration and prevention of side effects and risks, use during pregnancy, non-compliance, selection of patients, information and instruction, the question whether new and better prophylactic agents are about to oust lithium, the use of combination treatment, the effect of lithium on suicidal behavior, the benefits of prophylactic lithium treatment, and the design of future comparative trials. PMID- 11869751 TI - The Stanley Foundation Bipolar Treatment Outcome Network. I. Longitudinal methodology. AB - The NIMH-Stanley Foundation Bipolar Treatment Outcome Network, a multisite clinical trials network, has been established to address many of the neglected areas of research in bipolar illness. The Network was designed so that it would be able to conduct randomized clinical trials at several different levels of methodologic rigor (blinded and open-label) both in academic and community practice settings in order to better assess long-term efficacy of existing treatments and develop new ones. In this fashion, large numbers of representative patients with bipolar disorder have been enrolled with an additional focus of elucidating possible clinical and biological predictors of treatment response. The unique focus of the Network is its systematic longitudinal approach to illness so that patients can be assessed comprehensively over the long-term in sequential randomized clinical trials at critical clinical decision points where data on relative efficacy are inadequate. Bipolar I and bipolar II patients with a range of illness variants and comorbidities are included. Daily prospective ratings of severity of mania and depression and associated degree of functional impairment are completed on the NIMH-Life Chart Method and a modified Clinical Global Impressions Scale for Bipolar Illness (CGI-BP) is utilized. More detailed cross-sectional ratings for depression (Inventory of Depressive Symptomatology), mania (Young Mania Rating Scale), and psychosis (Positive and Negative Syndrome Scale) are additionally used at academic centers. This article describes the rationale for the Network, its guiding principles, methods, and study design to systematically assess the highly variable course of bipolar illness and its response to current and future treatments. PMID- 11869752 TI - The Stanley Foundation Bipolar Treatment Outcome Network. II. Demographics and illness characteristics of the first 261 patients. AB - BACKGROUND: Since recent NIMH Bipolar Disorder Workshops highlighted the dearth of longitudinal and controlled studies of bipolar illness, the Stanley Foundation Bipolar Network (SFBN) has recruited a large cohort of patients with bipolar disorder to begin to address these issues. This report describes the demographics and course of illness characteristics of this study population. METHODS: The first 261 outpatients to be diagnosed by the Structured Clinical Interview for DSM-IV (SCID) and complete a detailed patient and a brief clinician questionnaire are described. All patients met DSM-IV criteria for bipolar I (n=211), bipolar II (n=42), or NOS (n=5) or schizoaffective (n=3), bipolar type. Chi-square and t tests were used to examine statistically significant associations among important demographic and descriptive items. RESULTS: The general demographic and illness characteristics were similar to those in many bipolar clinical samples and not dissimilar from those reported in epidemiological surveys. The majority of patients had been hospitalized, with almost half reporting a worsening of illness over time, and two-thirds were not asymptomatic between episodes. First treatment for patients had been delayed by an average of 10 years from illness onset (by SCID). Almost a third of patients had attempted suicide at least once, and 30% reported current suicidal ideation at study entry. A total of 62% reported moderate to severe impact of the illness on occupational functioning. Early onset bipolar illness (< or =17 years old) was associated with increased frequency of mood switches, worsening course of illness, and history of early abuse (physical, verbal, or sexual). CONCLUSION: The SFBN represents a sample of predominantly BP I patients largely recruited from the community who will be followed in detail longitudinally, participate in clinical trials, and thus help advance our understanding and treatment of this life-threatening medical disorder. While there is a broad range of illness characteristics and severity, the majority of patients have been severely impacted by their illness despite the availability of multiple conventional treatment approaches in the community. These data further underscore the need for development of new and earlier treatment interventions. LIMITATION: The SFBN population is limited by the lack of random selection and represents a cohort willing to be treated and followed intensively in academic tertiary referral centers. While its characteristics are similar to many clinical study populations, the generalizability to non-clinic populations remains uncertain. PMID- 11869753 TI - Principles of effectiveness trials and their implementation in VA Cooperative Study #430: 'Reducing the efficacy-effectiveness gap in bipolar disorder'. AB - Despite the availability of efficacious treatments for bipolar disorder, their effectiveness in general clinical practice is greatly attenuated, resulting in what has been called an 'efficacy-effectiveness gap'. In designing VA Cooperative Studies Program (CSP) Study #430 to address this gap, nine principles for conducting an effectiveness (in contrast to an efficacy) study were identified. These principles are presented and discussed, with specific aspects of CSP #430 serving as illustrations of how they can be implemented in an actual study. CSP #430 hypothesizes that an integrated, clinic-based treatment delivery system that emphasizes (1) algorithm-driven somatotherapy, (2) standardized patient education, and (3) easy access to a single primary mental health care provider to maximize continuity-of-care, will address the efficacy-effectiveness gap and improve disease, functional, and economic outcome. It is an 11-site, randomized controlled clinical trial of this multi-modal, clinic-based intervention versus usual VA care running from 1997 to 2003. The trial has enrolled 191 subjects in each arm, using minimal exclusion criteria to maximize the external validity of the study. Subjects are followed for 3 years. The intervention is highly specified in a series of operations manuals for each of the three components. Several continuous quality improvement (CQI) interventions, process measures, and statistical techniques deal with drift of care in both the intervention and usual care arms to ensure the internal validity of the study. CSP #430 is designed to have impact well beyond the VA, since it evaluates a basic health care operational principle: that augmenting ambulatory access for major mental illness will improve outcome and reduce overall treatment costs. If results are positive, this study will provide a reason to reconsider the prevailing trend toward limitation of ambulatory services that is characteristic of many managed care systems today. PMID- 11869754 TI - The significance of psychotic features in manic episodes: a report from the NIMH collaborative study. AB - BACKGROUND: Psychotic features in the context of major depressive syndromes have correlates in symptom severity, acute treatment response and long-term prognosis. Little is known as to whether psychotic features have similar importance when they occur within manic syndromes. METHODS: These data derive from a multi center, long-term follow-up of patients with major affective disorder. Raters conducted follow-up interviews at 6-month intervals for the first 5 years and annually thereafter. A sub-set of probands participated in a family study in which all available, adult, first-degree relatives were interviewed as well. RESULTS: Of 139 who entered the study in an episode of mania, 90 patients had psychotic features. Symptom severity ratings at intake were more severe for this group. Though time to first recovery and time to first relapse did not distinguish the groups, psychotic features were associated with a greater number of weeks ill during follow-up and the strength of this association was similar to that seen for psychotic features within depressed patients described in an earlier publication. Patients with psychotic mania at intake did not differ significantly from those with nonpsychotic mania by response to acute lithium treatment, suicidal behavior during follow-up, or risks for affective disorder among first-degree relatives. Psychotic features within manic syndromes were not associated with high psychosis ratings during follow-up. In contrast, when psychotic features accompanied depressive syndromes, they strongly predicted the number of weeks with psychosis during follow-up, particularly among individuals whose episodes at intake were less acute. CONCLUSIONS: As with major depressive syndromes, psychotic features in mania are associated with greater symptom severity and higher morbidity in the long-term. Psychotic features are much less predictive of future psychosis when they occur within a manic syndrome than when they occur within a depressive syndrome. PMID- 11869755 TI - Toward a refined phenomenology of mania: combining clinician-assessment and self report in the French EPIMAN study. AB - BACKGROUND: Because manic patients lack insight, they are generally considered unreliable observers of their own psychopathology. The present analyses sought to examine to what extent patient reports could improve formal diagnostic criteria for mania--and be validated against the Carroll-Klein (CK) psychobiological model of bipolarity. METHOD: 104 DSM-IV acutely manic (hospitalized) patients provided self-assessment on the Ahearn--Carroll scale, the Multiple Visual Analogue Scales of Bipolarity (MVAS-BP). A principal component analysis (PCA) was performed on MVAS-BP, and the data on factorial scores were then compared to dimensional scores according to the CK model and to factors on the Beigel-Murphy Manic State Rating Scale (MSRS) completed by psychiatrists. RESULTS: The PCA identified a general factor accounting for 33% of the total variance; after varimax rotation, seven independent factors emerged, essentially in coherence with the signs and symptoms of DSM-IV mania, except for the 'social disinhibition' factor, which does not figure out as a distinct criterion in DSM-IV. Strong correlations were obtained (r > or = 0.80) between the four major factors of MVAS-BP and the four dimensional categories of the CK model: 'Consummatory Reward' with F1 'Elation and Inflated Self-esteem' (r=0.93), 'Incentive Reward' with F2 'Activation' (r=0.84), 'Psychomotor Pressure' with F3 'Acceleration' (r=0.85), and 'Central Pain' with F4 'Anxiety-Depression' (r=0.84). The F2 'Activation' appeared to be strongly correlated (r > or = 0.70) to all categories of the CK model. Correlational analysis between the factor structure of MVAS-BP and the MSRS showed significant coefficients on the scores assessing the emotional factors of 'Elation' and 'Depression.' Among the MVAS-BP factors, only 'Activation' was correlated to the majority of clinician ratings as obtained by the MSRS. CONCLUSIONS: These findings provide overall construct validity to the DSM-IV criteria for mania. Self-assessment of this disorder appears feasible and potentially useful in practice; lack of insight, poor judgment, and distractibility obviously require assessment by a clinician. Although our data are correlational and require prospective validation, they nonetheless suggest that (1) activation should be raised to the status of the stem criterion for mania, (2) to specify mood as elated, depressive, anxious, or irritable, and (3) to give individual status to social disinhibition (indiscriminate gregariousness) as a core pathological behavior in mania. Combining clinician- and self observation thus produces a more precise and complete phenomenology of mania. We finally submit that the foregoing reformulation provides a psychobiological basis to the manic construct as formulated in the Carroll-Klein model. PMID- 11869756 TI - The feasibility of self-assessment of dysphoric mania in the French national EPIMAN study. AB - BACKGROUND: There is presently considerable uncertainty on how to best assess mixed mania. The present contribution explores the feasibility of discriminating manic and dysphoric manic states on the basis of self-rating in the acute phase of the illness. METHODS: In the French four-site national EPIMAN study of 104 patients devoted to the clinical evaluation and subclassification of mania, we used the Multiple Visual Analog Scales of Bipolarity (MVAS-BP, 26 items) of Ahearn-Carroll in a self-assessment format. The study was conducted on consecutive patients hospitalized for an acute DSM-IV mania. The severity of mania was measured by the Beigel-Murphy scale (MSRS) assessed by psychiatrists. When mania abated, temperaments according to Akiskal and Mallya were administered in their French version. RESULTS: Principal component analysis revealed a general factor explaining 33% of the variance and, after rotation, seven factors defining different dimensions of the phenomenology of mania. The factorial scores, as well as the dimensional scores of the Carrol-Klein model significantly distinguished pure versus dysphoric mania made on clinical grounds. Gender seemed to influence two factors: high 'anxious-depressive' score in females (which is in line with female overrepresentation in mixed mania), vs. high score in males on the 'gregariousness' factor (which represents social disinhibition of the hyperthymic temperament known to be more prevalent in men). LIMITATION: Cross-sectional correlational study in need of longitudinal validation. CONCLUSIONS: EPIMAN data deriving from a national clinical population showed the feasiblity and face validity of self-assessment in acute mania, in particular its dysphoric subtype. Temperament in women seemed to contribute to the genesis of mixed (dysphoric) mania in accordance with Akiskal's hypothesis of opposition of temperament and polarity of bipolar episodes in mixed states. Self-assessment was capable of capturing accurately the subthreshold depressive symptomatology of mixed mania, which can be missed in hetero-evaluation by hasty clinical interview. PMID- 11869758 TI - Delineating bipolar II mixed states in the Ravenna-San Diego collaborative study: the relative prevalence and diagnostic significance of hypomanic features during major depressive episodes. AB - BACKGROUND: Depressive mixed state (DMX), defined by hypomanic features during a major depressive episode (MDE) is under-researched. Accordingly, study aims were to find DMX prevalence in unipolar major depressive disorder (MDD) and bipolar II depressive phase, to delineate the most common hypomanic signs and symptoms during DMX, and to assess their sensitivity and specificity for the diagnosis of DMX and bipolar II. METHODS: 161 unipolar and bipolar II MDE psychotropic drug- and substance-free consecutive outpatients were interviewed during an MDE with the Structured Clinical Interview for DSM-IV. DMX was defined at two threshold levels as an MDE with two or more (DMX2), and with three or more (DMX3) simultaneous intra-episode hypomanic signs and symptoms. RESULTS: DMX2 was present in 73.1% of bipolar II, and in 42.1% of unipolar MDD (P<0.000); DMX3 was present in 46.3% of bipolar II, and in 7.8% of unipolar MDD (P<0.000). The most common hypomanic manifestations during MDE were irritability, distractibility, and racing thoughts. Irritability had the best combination of sensitivity and specificity for the diagnosis of DMX2 and DMX3. Various combinations of irritability, distractibility, and racing thoughts correctly classified the highest number of DMX2 and DMX3, and had the strongest predictive power. DMX2 had high sensitivity and low specificity for bipolar II, whereas DMX3 had low sensitivity (46.3%) and high specificity (92.1%). LIMITATIONS: Single interviewer, cross-sectional assessment, and interviewing clinician not blind to patients' unipolar vs. bipolar status. CONCLUSIONS: When conservatively defined (>or = 3 intra-episode hypomanic signs and symptoms during MDE), DMX is prevalent in the natural history of bipolar II but uncommon in unipolar MDD. These findings have treatment implications, because of growing concerns that antidepressants may worsen DMX, which in turn may respond better to mood stabilizers. These data also have methodological implications for diagnostic practice: rather than solely depending on the vagaries of the patient's memory for past hypomanic episodes, the search for hypomanic features--ostensibly elation would not be one of those- during an index depressive episode could enhance the detection of bipolar II in otherwise pseudo-unipolar patients. Strict adherence to current clinical diagnostic interview instruments (e.g. the SCID) would make such detection difficult, if not impossible. PMID- 11869759 TI - Structure of mania: depressive, irritable, and psychotic clusters with different retrospectively-assessed course patterns of illness in randomized clinical trial participants. AB - BACKGROUND: We investigated the structure of manic episodes by determining whether there was evidence for distinct groups of patients differing in clinical characteristics and course of illness. METHODS: The subjects were 162 patients hospitalized for manic episodes who underwent comprehensive evaluations of behavior, symptoms, and history before a treatment study. Pretreatment behavior ratings (Schedule for Affective Disorders and Schizophrenia, rated by clinicians, and Affective Disorder Rating Scale, rated by nurses) entered a principal components factor analysis, followed by a cluster analysis of the subjects based on their factor scores. Members of the resulting clusters were compared with respect to clinical characteristics and history of illness. RESULTS: The six factors were impulsivity, hyperactivity, anxious pessimism, distressed appearance, hostility, and psychosis. The four clusters were characterized as depressive, with high anxious pessimism (n=22), delusional, with high psychosis (n=39), classic (n=72), and irritable, with high distressed appearance and hostility (n=29). Depressive manics had the earliest onset of illness and the highest density of episodes/year, while irritable manics had later onset and the fewest previous episodes. LIMITATIONS: The number of subjects was smaller than ideal for multivariate analysis, subjects were limited to those able to consent and meet criteria for a randomized clinical trial, and course of illness was determined retrospectively. CONCLUSIONS: Manic episodes have a dimensional structure but appear to fall naturalistically into types that differ with respect to previous history, symptoms, and clinical characteristics. Whether these are distinct clinical subtypes will require further research. PMID- 11869757 TI - Clinical characterization of depressive mixed state in bipolar-I patients: Pisa San Diego collaboration. AB - BACKGROUND: Although mixed states were classically described as various concomitant admixtures of depression and mania, the official current definitions in both DSM-IV and ICD-10 tend to restrict the concept to manic patients with full syndromal depression. Recent research has actually shown that mania with few depressive symptoms constitutes the most prevalent clinical presentation of mixed or dysphoric mania. Major depressive patients with few concomitant manic symptoms are not officially recognized within the current nosology. In this paper we attempt to delineate the clinical profile of such depressive mixed states in the context of bipolar I disorder. METHODS: In the Pisa day center, we studied 195 bipolar I patients who either met Pisa criteria for bipolar mixed state (n=159) or DSM-III-R criteria for major depressive episode (bipolar major depression or B MD, n=36). Of the 159 patients identified by Pisa criteria as mixed state, 86 also met the criteria of the DSM-III-R for mixed episode (core mixed state or MS group), while 32 met the DSM III-R criteria for major depressive episode (provisionally defined as depressive mixed states, D-MS); the remaining patients (n=41, 25.7%) with predominatly manic picture were not included in the present comparisons. RESULTS: The three groups (B-MD, MS and D-MS) had close similarities in clinical and sociodemographic characteristics such as age, sex distribution, marital status, schooling, residence, age at onset, age of first treatment, age of first hospitalization, degree of chronicity of the index episode, stressor within the 6 months before the index episode, lifetime suicide attempts and premorbid temperament. First degree family history for bipolar illness and that for other mental disorders was also similar, except for major depression that was more common among the relatives of D-MS. MS and D-MS were further distinguished from B-MD by the fact that the latter followed a more 'cyclic' course with shorter yet greater number of episodes, and which began with a pure depressive episode; by contrast, MS and D-MS had fewer episodes of longer duration, less interepisodic remission, and tended to begin with a mixed episode. Incongruous psychotic features were more common in the two mixed groups compared to B-MD, and the most common features of the D-MS group were agitation, psychotic depression with irritable mood, pressured speech and/or flight of ideas. LIMITATION: It was not feasible to collect information blind to clinical status in patients with severe psychotic mood states. CONCLUSION: These data confirm the existence of psychotic agitated depressive mixed states with flight of ideas, distinct from cyclic retarded pure bipolar depressive states. The recognition of these affective states is clinically important to protect patients from the potentially harmful indiscriminate use of antidepressants and to provide them with the benefits of an anticonvulsant, a short-term neuroleptic, or ECT. PMID- 11869760 TI - Olanzapine versus haloperidol in schizoaffective disorder, bipolar type. AB - BACKGROUND: The present analysis was performed on data from a subsample of patients with schizoaffective disorder, bipolar type, who participated in a multicenter, double-blind study comparing olanzapine to haloperidol. METHODS: Patients with schizoaffective disorder bipolar type, characterized as currently manic, mixed, depressed, or euthymic, were assessed weekly for 6 weeks during treatment with either olanzapine or haloperidol. Manic symptoms were measured using the sum of six items of the BPRS, and depressive symptoms were assessed using the Montgomery-Asberg Depression Rating Scale. In addition, cognitive functioning was measured using the sum of seven items from the PANSS. Repeated measures analyses were performed using random coefficients regression of the serial measurement of manic, cognitive, and depressive symptoms. RESULTS: A significant treatment difference was detected overall, indicating that olanzapine was significantly more effective than haloperidol in reducing symptoms of depression and improving patients' cognitive symptoms. The superiority of olanzapine over haloperidol in the reduction of manic symptoms did not reach statistical significance (P=.052). The greatest improvement in both manic and cognitive symptoms was seen in the olanzapine-treated 'currently manic' subgroup, and least improvement in the haloperidol-treated 'euthymic' subgroup. Depressive symptoms were most improved in the olanzapine-treated 'depressed' subgroup, and least improved in the corresponding haloperidol subgroup. CONCLUSIONS: Overall, olanzapine was superior to haloperidol with respect to thymoleptic effects in patients with schizoaffective disorder, bipolar type. PMID- 11869762 TI - Impact of bipolar affective disorder on family and partners. AB - BACKGROUND: Successful management of major mental illness in the community relies significantly on an informal or non-professional network of caregivers. The needs and experiences of such caregivers have been little studied with respect to major chronic mood disorders. METHOD: A sample of caregivers (n=41) of RDC bipolar disorder was systematically interviewed to determine how this role affected them. RESULTS: Caregivers reported significant difficulties in their relationships with the patient when s/he was unwell, with considerable impact on their own employment, finances, legal matters, co-parenting and other social relationships. Violence was a particular worry for partner/parent caregivers of both male and female patients when the patient was manic. The caregiver's own mental health appeared unaffected. Despite this, the caregivers appeared emotionally committed to the patients and showed considerable tolerance of problem behaviours, which they rank-ordered for difficulty. Among nonfamily partners, knowledge of the illness before cohabitation was poor. LIMITATION: The sampling does not capture caregivers who have abandoned their role, such as spouses who have divorced the bipolar sufferer. CONCLUSIONS: Management of this illness requires a partnership between mental health professionals and the informal caregivers and the authors suggest that each group needs to understand the difficulties encountered by the other. Although erosion of relationships is a well-known complication of bipolar disorder, findings indicate that treating clinicians can rely on caregivers committed to the welfare of the patient. PMID- 11869761 TI - Are antidepressants less effective in the acute treatment of bipolar I compared to unipolar depression? AB - BACKGROUND: Using our routine documentation system we evaluated the hypothesis that antidepressants may be less effective in the acute treatment of bipolar I depressed patients compared to unipolar depressed patients. METHOD: Based on the data from 2032 consecutively admitted inpatients with unipolar or bipolar I depression we compared the efficacy of antidepressants in both groups. The outcome was assessed by the Global assessment scale (GAS), the duration of hospitalisation and the Apathic-, Depressive- and Manic Syndrome subscales of the Association for Methodology and Documentation in Psychiatry system. RESULTS: Cohorts were comparable in treatment regimens and severity of depression at admission. At discharge, there were no statistically significant differences between bipolar I and unipolar depression for the outcome criteria Depressive Syndrome scale, GAS score and days in hospital. Bipolar patients showed a slightly decreased score of the Apathic Syndrome scale at discharge, also reflected by a slightly elevated score of the Manic Syndrome scale. LIMITATIONS: This study did not check for differences in side effects such as switching or cycling commonly attributed to the use of tricyclic antidepressants in bipolar patients. Naturalistic data were obtained prospectively, analysis, however, was done retrospectively, thus limiting, but not completely excluding a possible selection bias. CONCLUSION: Evaluating different outcome criteria, our naturalistic data of a large cohort seem to reject the hypothesis that antidepressants may be less effective in the acute treatment of bipolar I compared to unipolar depressed patients. PMID- 11869763 TI - Measures of attention and hyperactivity symptoms in a high-risk sample of children of bipolar parents. AB - BACKGROUND: To determine whether significant symptoms of inattention were present among the offspring of well-characterized bipolar parents. METHODS: We included 53 offspring of 30 parents meeting DSM-IV criteria for bipolar disorder diagnosed by consensus on the basis of a SADS-L interview and a wealth of longitudinal clinical data. The unaffected parent had no lifetime history of a major psychiatric illness. Offspring, prospectively followed for up to 5 years, completed psychometric measures of attention and mood when judged to be at a good level of functioning (well, remitted or treated). RESULTS: Those offspring with any lifetime psychiatric diagnosis endorsed more subjective problems with attention. However, there was no measurable difference on tasks of sustained attention between those with and those without a lifetime psychiatric illness including affective disorder. There was a significant association between self reported symptoms of depression and inattention, but no association between either self-report measure and an objective measure of sustained attention. LIMITATIONS: This study was not intended to be a comprehensive neuropsychological investigation of at risk offspring. CONCLUSIONS: In this high-risk population, subjective difficulty with attention appeared to be state-dependent, associated with the degree of subjective distress related to an underlying psychiatric illness. PMID- 11869764 TI - Anxiety disorders comorbidity in mood disorder subgroups: data from a mood disorders clinic. AB - BACKGROUND: Previous research has identified a high rate of anxiety disorders comorbidity in patients with a primary mood disorder diagnosis. Discrepancies between studies in the comorbidity prevalence of specific anxiety disorders in mood disorders, and of anxiety disorders comorbidity between unipolar depression and bipolar mood disorder are in part due to differences in sampling and diagnostic assessment methodology. METHOD: The authors reviewed the charts of 138 patients who received the SCID-P for DSM-III on enrollment in a Mood Disorders Clinic during the period 1982 through 1988. The comorbidity of specific DSM-III Anxiety Disorders with specific mood disorders was determined and comparatively examined using non-parametric statistics. RESULTS: There was high overall comorbidity of anxiety disorders that did not differ between bipolar and unipolar subjects. There were no differences in the comorbidity of individual anxiety disorder diagnoses in the unipolar vs. bipolar groups. However, in unipolar patients with, compared to those without an additional diagnosis of dysthymia, there was greater overall anxiety disorders comorbidity, with a particularly high prevalence of generalized anxiety disorder. LIMITATION: The subgroup of patients with bipolar I disorder was relatively small (N=8). CONCLUSION: Mood and anxiety disorders comorbidity is complex and presents a continuing challenge for both clinicians and researchers. PMID- 11869765 TI - Anxiety disorders comorbidity in bipolar I, bipolar II and unipolar major depression: results from a population-based study in Hungary. AB - BACKGROUND: The aim of this study was to analyze the lifetime comorbidity between DSM-III-R anxiety disorders in separate subgroups of patients with major depression, bipolar II and bipolar I disorder in a community sample of a Hungarian population. METHODS: Randomly selected subjects (aged between 18 and 64 years, N=2953) were interviewed by the Diagnostic Interview Schedule (DIS) which generated DSM-III-R diagnoses. RESULTS: The prevalence of generalized anxiety disorder, agoraphobia and simple phobia was the highest among bipolar II patients (20.8, 37.5 and 16.7%, respectively), social phobia was most prevalent in (nonbipolar) major depression (17.6%), while the rate of panic disorder was the same in the (nonbipolar) major depressive and bipolar II subgroups (12.4 and 12.5%, respectively). Bipolar I patients showed a relatively low rate of comorbidity. CONCLUSIONS: The findings support previous results on the particularly high rate of lifetime comorbidity between anxiety disorders and unipolar major depression and particularly bipolar II illness. LIMITATIONS: Underestimation of the prevalence of bipolar II disorder by the diagnostic methodology used, resulting in a small number of bipolar II cases, lack of analysis of data by gender, no data on obsessive-compulsive disorder. PMID- 11869766 TI - A prospective study of inter-episode consistency of manic and mixed subtypes of bipolar disorder. AB - BACKGROUND: Few studies have compared symptom presentations across manic or mixed episodes in manic-depressive patients. METHODS: In the current study we report on symptom presentations of 68 prospectively-evaluated subjects diagnosed with Bipolar Disorder during two discrete manic or mixed episodes. Each episode was categorized using DSM-IIIR criteria for Bipolar Disorder, manic or mixed, as well as a less restrictive definition for manic and mixed states derived from receiver operating characteristic (ROC) analysis of symptoms. RESULTS: The occurrence of mixed bipolar episodes was not random using either the DSM-IIIR or ROC-derived definitions of mixed episodes. LIMITATIONS: Subjects were not all fully medication-free at the time of evaluation which may have altered symptom presentation. The total duration of the study was limited, with the longest inter episode interval under 6 years. CONCLUSIONS: Although there was variability in mixed symptomatology between episodes, the occurrence of mixed episodes was not random. Manic and mixed episodes tend to recur true to type. PMID- 11869767 TI - Unsuspected depressive mania in pre-pubertal Hispanic children referred for the treatment of 'depression' with history of social 'deviance'. AB - BACKGROUND: Despite an emerging Literature on the mixed nature of pediatric mania, initial presentation with conduct problems continues to mislead mental health clinicians. The present report focuses on Hispanic pre-pubertal children referred for the treatment of depression in the context of conduct problems. METHODS: Eleven boys and two girls received a structured psychiatric assessment in a practice setting to make sense of the presenting clinical complexity. Diagnoses were assigned using the DSM-IV criteria. RESULTS: Ten of the boys and both girls met criteria for depressive mania. Their family histories were replete with affective disorder. Five (50%) of the boys and both of the girls (100%) with depressive mania had family histories of bipolar disorder. Six (60%) of the boys and neither of the girls with depressive mania had psychotic features. Those with depressive mania exhibited clear-cut circadian changes in symptomatology. Euphoria, oscillating with affective states indicative of psychic pain, was characteristically restricted to the evenings or nighttime. However, the drive to seek treatment had stemmed from social 'deviance'. CONCLUSION: Children with depressive mania are often unrecognized in clinical settings. Boys with conduct problems may be disproportionately represented among such children. These data support Akiskal's hypothesis that externalizing (conduct) problems in clinically referred children with depression are indicative of bipolar disorder. PMID- 11869768 TI - Valproate in very young children: an open case series with a brief follow-up. AB - We report nine cases of juvenile mania, of which six began in the preschool years. We provide validation by clinical description, family history of bipolar disorder, worsening on stimulants, and considerable mood stabilization with divalproex. This is a relatively new area of clinical observation, and systematic studies are needed to firmly establish this diagnostic category in very young children. Our case series enriches the existing scant literature and provide the rationale for the use of mood stabilizers rather than stimulants in this juvenile population. However, no controlled studies exist on the efficacy and safety of valproate in this age group; lithium that has received greater clinical attention, has not been subjected to controlled studies either. Our clinical observations with divalproex are preliminary but encouraging. PMID- 11869769 TI - The temporal relationship between anxiety disorders and (hypo)mania: a retrospective examination of 63 panic, social phobic and obsessive-compulsive patients with comorbid bipolar disorder. AB - BACKGROUND: The relationship between anxiety and depressive disorders has been conventionally limited to unipolar depression. Recent studies from both clinical and epidemiologic samples have revealed intriguing associations between anxiety and bipolar (mainly bipolar II) disorders. The present report examines the temporal sequence of hypomania to panic (PD), obsessive-compulsive (OCD) and social phobic (SP) disorders. METHODS: Specialty-trained clinicians retrospectively evaluated the foregoing relationships in 63 patients meeting the DSM-III-R diagnosis for PD, OCD and SP with lifetime comorbidity with bipolar disorders (87% bipolar II). Structured interviews were used. RESULTS: In nearly all cases, SP chronologically preceded hypomanic episodes and disappeared when the latter episodes supervened. By contrast, PD and OCD symptomatology, even when preceding hypomanic episodes, often persisted during such episodes; more provocatively, nearly a third of all onsets of panic attacks were during hypomania. LIMITATIONS: Assessing temporal relationships between hypomania and specific anxiety disorders on a retrospective basis is, at best, of unknown reliability. The related difficulty of ascertaining the extent to which past antidepressant treatment of anxiety disorders could explain the anxiety-bipolar II comorbidity represents another major limitation. CONCLUSIONS: Different temporal relationships characterized the occurrence of hypomania in individual anxiety disorder subtypes. Some anxiety disorders (notably SP, and to some extent OCD) seem to lie on a broad affective continuum of inhibitory restraint vs. disinhibited hypomania. By contrast, and more tentatively, PD in the context of bipolar disorder, might be a reflection of a dysphoric manic or mixed hypomanic symptomatology. The foregoing suggestions do not even begin to exhaust the realm of possibilities. The pattern of complex relationships among these disorders would certainly require better designed prospective observations. PMID- 11869770 TI - Divalproex therapy in medication-naive and mood-stabilizer-naive bipolar II depression. AB - BACKGROUND: There have been few systematic studies of the treatment of bipolar II depression. While divalproex sodium (DVPX) is effective in acute mania, there are few data on the antidepressant effects of DVPX. Similarly, little is known regarding the use of DVPX administered in a single daily dose. METHOD: We performed a 12-week open trial of DVPX monotherapy (mean dose 882 mg qhs, mean level 80.7 mug/ml) in nineteen (thirteen women, six men, mean age 29) bipolar II depressed outpatients. Eleven patients (six women, five men) were medication naive (MN) and eight (seven women, one man) were mood stabilizer-naive (MSN), having had prior trials of antidepressants or stimulants. Mean illness and current depressive episode duration were 15.4 years and 11.8 weeks, respectively. DVPX was given as a single dose each evening starting with 250 mg at bedtime and increased by 250 mg at bedtime every 4 days until symptom relief or adverse effects were noted. Weekly prospective Hamilton Depression, Young Mania and Clinical Global Impression ratings were obtained. RESULTS: DVPX therapy was generally well tolerated. Twelve of nineteen patients (63%) responded (>50% decrease in Hamilton Depression ratings). MN patients compared to MSN patients tended to have a higher response rate (9/11 versus 3/8, P<0.08). Mean Hamilton scores decreased from 22.2 to 9.6 (P<0.0001) in the entire group, from 20.6 to 6.6 (P<0.0003) in MN patients, and from 24.2 to 14.7 (P=0.008) in MSN patients. CONCLUSION: Single daily dose DVPX monotherapy appeared to be well tolerated and substantially benefited 63% of patients with bipolar II depression. The trend towards a higher rate of antidepressant response to DVPX in MN patients (82%) compared to MSN patients (38%) could be due to a milder form or earlier phase of illness and the lack of prior medication exposure or failures. This uncontrolled open pilot study must be viewed with caution, and randomized double-blind placebo controlled studies of DVPX in bipolar II depression are warranted to confirm the possibility that single daily dose DVPX is an effective, well-tolerated, first line monotherapy in this population. PMID- 11869771 TI - Role of risperidone in bipolar II: an open 6-month study. AB - BACKGROUND: Since treatment approaches thought to be useful for mania are presumably suitable for hypomania as well, little systematic research has been done on the treatment of hypomanic episodes and their long-term outcome. As systematic trials have shown that the atypical antipsychotic risperidone may be effective and safe in the treatment of acute mania, we decided to conduct an open label study of its effectiveness and tolerability in hypomania associated with bipolar II. METHODS: Forty-four DSM-IV bipolar II patients with Young Mania Rating Scale (YMRS) scores above 7 were included and followed-up for 6 months. Efficacy was measured by means of the YMRS and the Clinical Global Impression for Bipolar Disorder (CGI-BD). Treatment-emergent depression was measured by the Hamilton Depression Rating Scale (HDRS-17), and the Udvalg for Kliniske Undersogelser (UKU) subscale was used for neurological/extrapyramidal side effects. RESULTS: Thirty-four patients completed the trial. The mean dose of risperidone at endpoint was 2.8 mg/day. Last observation-carried-forward analysis showed significant reduction of YMRS scores from the first week of treatment, which continued until the endpoint (P<0.0001). At 6-month follow-up, 60% of patients were assymptomatic according to the CGI. The 32% who received risperidone in monotherapy seemed to respond equally well. Risperidone, as used in this study, appeared to be most protective against hypomanic than depressive recurrences. Nine patients (12%) had a depressive relapse during 6-month follow up, one patient (2%) had an hypomanic relapse and another (2%) had both. No patients developed tardive dyskinesia during the duration of the study. Although most patients received risperidone in combination with standard mood-stabilizers, only three patients discontinued risperidone because of other side-effects. LIMITATIONS: In the absence of a placebo arm, it is uncertain to what extent the foregoing results could be ascribed to spontaneous remission of bipolar II disorder. CONCLUSIONS: Risperidone, either in combination with mood-stabilizers or alone was well-tolerated in bipolar II patients, who presented in a hypomanic state, and appeared efficacious. Further controlled research on the role of atypical antipsychotics in the treatment of less-than-manic forms of bipolar illness is warranted. PMID- 11869772 TI - Do patients with borderline personality disorder belong to the bipolar spectrum? AB - BACKGROUND: This report examines clinical indicators for bipolarity in a cohort of patients suffering from Borderline Personality Disorder (BPD). METHODS: The study was conducted in the Cornell-Westchester Hospital, famed for its expertise in BPD. To avoid biasing our sample, we excluded all BPD patients who were active patients in our anxiety and mood disorders program. Through the use of both open clinical interviews and standardized diagnostic interviews (SCID), borderline patients were examined for evidence of bipolarity by five indicators: history of spontaneous mania, history of spontaneous hypomania, bipolar temperaments, pharmacologic response typical of bipolar disorder, and a positive bipolar family history. RESULTS: Depending on the level of bipolar disorder from the most rigorous (mania) to the most 'soft' (bipolar family history), between 13 and 81% of borderline patients showed signs of bipolarity. Based on what the emerging literature supports as rigorously defined bipolar spectrum (bipolar I and II), we submit that at least 44% of BPD belong to this spectrum; adding hypomanic switches during antidepressant pharmacotherapy, the rate of bipolarity in BPD reaches 69%. As expected from this formulation, most responded negatively to antidepressants (e.g. hostility and agitation) and positively to mood stabilizers. LIMITATIONS: Small sample size and retrospective gathering of data on treatment response. CONCLUSION: Patients with BPD more often than not exhibit clinically ascertainable evidence for bipolarity and may benefit from known treatments for Bipolar Spectrum Disorders. Large scale, systematic treatment studies with mood stabilizers are indicated. PMID- 11869773 TI - Origin and development of concepts of bipolar mixed states. AB - This paper reviews the historical origins of the contemporaneous resurgence of interest in mixed states. This is a classical concept whose origins can be traced back to ancient times. In more modern times, already in the pre-Kraepelinian era we can find descriptions and classifications of "mixed states". For example, in his classification of mental disorders described "mixtures of exaltations and depression", and he distinguished among "mixed mood disorders", "mixed mental disorders", and "mixed volition disorders". Subsequently, (the father of empirical and biological psychiatric research in Germany) described the "mid forms". Half a century later we encounter the crucial role of Emil Kraepelin and the development and systemization of his views between 1899 and 1913--leading to the characterization of such conditions as "depressive-anxious mania", "excited depression", and "stuporous mania". The remainder of this article focuses on the essential points of the first book on mixed states in the psychiatric literature: On The Mixed States of Manic-Depressive Insanity by. For much of the present 20th century nothing new emerges, followed by a contemporary renaissance of mixed states, particularly in the United States. The paper concludes with proposal of mixed states as temperament intruding into an episode of opposite polarity. PMID- 11869774 TI - Current research on rapid cycling bipolar disorder and its treatment. AB - Rapid cycling is a pattern of presentation of bipolar disorder that specifies the course of the illness and is associated with a greater morbidity. The validity of rapid cycling as a distinct course modifier for bipolar disorder has been demonstrated and the term has been incorporated into the DSM-IV. The phenomenon of rapid cycling tends to appear late in the course of the disorder, occurs more frequently among females, and is more frequently seen in patients with bipolar type II disorder. Stimulants such as cocaine may also play some role in rapid cycling. It is generally accepted that a recent history of rapid cycling predicts non-response to monotherapy with lithium and probably carbamazepine as well; however it is also possible that concurrent use of antidepressants may play a role in destabilizing the illness course under these agents. Thus, clinical considerations suggest that discontinuing antidepressants may facilitate the recovery process. Among clinically available monotherapies, valproate and lamotrigine appear to be the most useful clinically. However, other treatments such as lithium, carbamazepine, the atypical antipsychotic agents, thyroid hormone, and bupropion are frequently needed augmentation strategies. Electroconvulsive therapy may also prove efficacious in selected cases. The present paper provides a critical review of the evidence for the foregoing clinical issues in rapid cycling. PMID- 11869775 TI - Clinical correlates of therapeutic response in bipolar disorder. AB - Interest in the factors associated with responsiveness to therapy in bipolar disorder has increased with evidence that lithium has differential rather than uniform effectiveness in bipolar disorder, and also with the broadening of therapeutic options, illustrated especially by the use of divalproex sodium (valproate). Lithium is effective acutely in patients with pure or elated mania, and in its prophylaxis, but may worsen depressive symptoms in such patients when used for long-term maintenance therapy. The nature of previous responses to lithium treatment predicts the type of response that can be expected during a further episode of the disorder. Mixed mania, secondary mania, and mania associated with substance abuse--as well as rapid cycling --generally respond poorly to lithium therapy. Divalproex sodium has a broader spectrum of efficacy and fewer factors that determine differential responsiveness. Data related to treatment with carbamazepine are sparse, but patients with rapid cycling may respond less well to this agent; on the other hand, limited data support the utility of carbamazepine in bipolar patients with extreme psychosis, including those with mood-incongruent features. Both classical and atypical neuroleptics (e.g. olanzapine) could profitably be used in acute mania and mixed states. Lamotrigine has recently been shown effective in bipolar depression and rapid cycling. Patients unresponsive to a single agent may show improvement with combined regimens, though this impression is based more on clinical experience than controlled randomized assignment. PMID- 11869776 TI - Genetic linkage and bipolar disorder: a cautionary note. PMID- 11869778 TI - The comorbidity of bipolar and anxiety disorders: prevalence, psychobiology, and treatment issues. AB - BACKGROUND: Although symptoms of anxiety as well as anxiety disorders commonly occur in patients with bipolar disorder, the pathophysiologic, theoretical, and clinical significance of their co-occurrence has not been well studied. METHODS: The epidemiological and clinical studies that have assessed the overlap of bipolar and anxiety disorders are reviewed, with focus on panic disorder and obsessive-compulsive disorder (OCD), and to a lesser extent, social phobia and post-traumatic stress disorder. Potential neural mechanism and treatment response data are also reviewed. RESULTS: A growing number of epidemiological studies have found that bipolar disorder significantly co-occurs with anxiety disorders at rates that are higher than those in the general population. Clinical studies have also demonstrated high comorbidity between bipolar disorder and panic disorder, OCD, social phobia, and post-traumatic stress disorder. Psychobiological mechanisms that may account for these high comorbidity rates likely involve a complicated interplay among various neurotransmitter systems, particularly norepinephrine, dopamine, gamma-aminobutyric acid (GABA), and serotonin. The second-messenger system constituent, inositol, may also be involved. Little controlled data are available regarding the treatment of bipolar disorder complicated by an anxiety disorder. However, adequate mood stabilization should be achieved before antidepressants are used to treat residual anxiety symptoms so as to minimize antidepressant-induced mania or cycling. Moreover, preliminary data suggesting that certain antimanic agents may have anxiolytic properties (e.g. valproate and possibly antipsychotics), and that some anxiolytics may not induce mania (e.g. gabapentin and benzodiazepines other than alprazolam) indicate that these agents may be particularly useful for anxious bipolar patients. CONCLUSIONS: Comorbid anxiety symptoms and disorders must be considered when diagnosing and treating patients with bipolar disorder. Conversely, patients presenting with anxiety disorders must be assessed for comorbid mood disorders, including bipolar disorder. Pathophysiological, theoretical, and clinical implications of the overlap of bipolar and anxiety disorders are discussed. PMID- 11869779 TI - Seasonal variation of mixed and pure episodes of bipolar disorder. AB - BACKGROUND: Although seasonal patterns of manic episodes have been reported, the seasonal variation of mixed states of bipolar disorder has received little attention. In the current report we address that concern as well as the overall seasonality of manic episodes. METHODS: The seasonal pattern of 304 psychiatric hospital admissions for treatment of mixed or manic bipolar episodes over a 3 year period were analyzed employing two definitions of mixed manic states: DSM III-R and an ROC derived definition. RESULTS: The frequency of all manic episodes combined peaked in early spring, with a nadir in late fall. Pure manic admissions showed a similar pattern. Mixed manic admissions had a significantly different pattern, with a peak in late summer and a nadir in November. The differences between pure and mixed manic admissions were demonstrated with the use of the ROC definition for mixed states. LIMITATIONS: Effects of medications and medication non-compliance may dampen natural seasonal patterns of episodes. CONCLUSIONS: The different seasonal pattern of mixed and pure manic episodes support the separation of mixed episodes as a distinct clinical subtype. PMID- 11869780 TI - Alcohol abuse in social phobic patients: is there a bipolar connection? AB - BACKGROUND: Epidemiological and clinical studies have reported the frequent co occurrence of social phobia (SP) and alcohol use disorders. Patients with SP often use alcohol to cope with the social situations they fear, and to lessen anticipatory anxiety, behavioral inhibition, and phobic avoidance. We investigated whether the presence of lifetime comorbidity with alcohol abuse was associated with significant differences as regards demographic and clinical features, family history and pattern of comorbidity in a large clinical sample of SP outpatients. METHOD: The sample comprised 153 outpatients who met DSM-III-R diagnostic criteria for SP. Demographic, family history and course characteristics were investigated by a semi-structured interview. Social phobic symptoms and the severity of the illness have been assessed by the Liebowitz Social Anxiety Scale (LSAS) and the Liebowitz Social Phobic Disorders Rating Scale, Severity (LSPDRS). Patients completed the Hopkins Symptom Checklist (HSCL 90). RESULTS: Thirty-four patients (22.2%) had a past or current history of alcohol abuse for at least 1 year. There were no significant differences between these patients and those without a history of alcohol abuse, as regards demographic features and lifetime comorbidity with major depression and other anxiety disorders. Bipolar disorder type II was found almost exclusively among patients with alcohol abuse, as well as family history for bipolar disorders. LIMITATIONS: Retrospective study. CONCLUSIONS: Our data indicate a strong relationship between bipolar II disorder and alcohol abuse comorbidity in patients with SP. The socializing and disinhibiting effect that many social phobics report might be mediated by mood elation induced by alcohol. The presence of bipolar diathesis in patients presenting with social anxiety might explain their increased susceptibility to alcohol, as they might undertake alcohol abuse as an attempt to overcome social difficulties. PMID- 11869781 TI - Psychopathological dimensions of depression: a factor study of the 17-item Hamilton depression rating scale in unipolar depressed outpatients. AB - BACKGROUND: Agreement on the factor structure of the Hamilton Depression Rating Scale (HDRS) has not been consistent among studies, and some investigators argued that the scale's factor structure is not reliable. This study aimed at shedding more light on this debated issue. METHODS: We studied 186 adults with unipolar depression (Major Depressive Disorder, n=80; Dysthymic Disorder, n=71; Depressive Disorder Not Otherwise Specified, n=25; Adjustment Disorder, n=10). They had no comorbid DSM-IV axis I or axis II disorders, and had received no treatment with antidepressant drugs in the previous 2 months. The factor structure of the scale was studied using the principal factor method, followed by oblique rotation. Factor scores were computed for each subject using the regression method. RESULTS: Using the scree-test criterion for factor extraction, we obtained a four factor solution, explaining 43.8% of total variance. The four factors extracted were identified as (1) somatic anxiety/somatization factor; (2) a psychic anxiety dimension; (3) a pure depressive dimension; and (4) anorexia factor. Patients with Major Depressive Disorder scored significantly higher than patients with other diagnoses on the pure depressive dimension. LIMITATIONS: These results need to be replicated in different cultures, using analogous factoring techniques. CONCLUSIONS: Though not exhibiting factorial invariance in the stricter sense of the term, the 17-item HDRS did exhibit a relatively reliable factor structure. Our analysis provides further evidence that the scale is multidimensional. However, as long as the multidimensional character of the scale is taken into account the scale should be able to play a useful role in clinical research. PMID- 11869782 TI - Adolescent life events as predictors of adult depression. AB - BACKGROUND: Among adults, life events predict future episodes of major depression as well as a range of anxiety disorders. While studies have begun to examine this issue in adolescents, few studies rely upon prospective epidemiological designs to document relationships between adolescent life events and adult major depression. METHOD: An epidemiologically-selected sample of 776 young people living in Upstate New York received DSM-based psychiatric assessments and an assessment of life events in 1986. Psychopathology was again assessed in 1992. The current study examined the predictive relationship between life events in 1986 and depression as well as anxiety in 1992, controlling for depression/anxiety in 1986. RESULTS: Adolescent life events predicted an increased risk for major depression diagnosis in adulthood. When analyzed continuously, an association emerged with symptoms of major depression as well as with symptoms of generalized anxiety disorder. However, this association with generalized anxiety disorder was limited to females. CONCLUSIONS: Life events in adolescence predict risk for major depression during early adulthood. PMID- 11869783 TI - Does SSRI augmentation with antidepressants that influence noradrenergic function resolve depression in obsessive-compulsive disorder? AB - BACKGROUND: Obsessive compulsive disorder (OCD) often coexists with major depressive disorder (MDD). Serotonergic antidepressant medications have emerged as the treatment of choice for both OCD and MDD. In the usual course of events, both the patient's OCD and depressive symptoms improve in parallel following initiation of serotonin reuptake inhibitor (SRI) treatment for OCD. However, such is not always the case. We report here on a series of ten patients whose OCD but not depression improved following a trial of SRI therapy. METHOD: Ten patients with OCD and comorbid MDD who experienced a worsening or exacerbation of depressive symptoms while being maintained on an adequate dose of SRI therapy were treated using a combination of SRIs and agents with effects on noradrenergic reuptake. Response to treatment was based on clinician-ratings of severity and improvement of OCD and MDD (CGI-S and CGI-I). RESULTS: Following augmentation, nine of the ten patients had a significant improvement/resolution of their MDD, with little further change in the severity of their OCD. LIMITATIONS: Inferences from the results of this study are limited by the lack of a control group, the small sample size, and the use of nonstandardized ratings as measures of symptom severity. CONCLUSIONS: These results are of practical significance to clinicians insofar as they suggest a possible guideline to clinicians treating depression in OCD with SSRIs without success. PMID- 11869784 TI - Color sensitivity and mood disorders: biology or metaphor? AB - BACKGROUND: A familiar but overlooked symptom in affective disorders is patient self-report of alterations in color sensitivity. Anecdotal and empirical evidence have suggested an association between mood and color sensitivity. The purpose of this pilot study was to test three hypotheses concerning the relationship between mood disorders and color sensitivity. METHODS: Using a cross-sectional survey design consisting of a sample of 120 inpatients and outpatients, color sensitivity was assessed by the patient's response to a self-report depression scale item, "I notice that everything seems gray/cloudy/drab/lacking color". RESULTS: Color sensitivity significantly correlated with depression in the total sample (P=0.001). The other two hypotheses approached significance but were not supported. DISCUSSION: These findings suggest there is evidence that color sensitivity is impaired during depression. Further research using a larger, more homogeneous sample and longitudinal design whereby measures of mood and color sensitivity are correlated before, during, and after treatment in depressed and manic patients would be justified. A study using ophthalmological instrumentation to measure color sensitivity would provide objective, 'hard' evidence of the association between color sensitivity and depression. CONCLUSIONS: Whether color perception is metaphorically reported by patients to describe their mood or a biological phenomenon remains to be validated. Findings seem to lend support to the conclusion that abnormalities in brain function alter retinal function. PMID- 11869785 TI - Late-onset depression: can EEG abnormalities help in clinical sub-typing? AB - BACKGROUND: The purpose of this study was to determine the frequency of EEG abnormalities indicative of mild cerebrovascular dysfunction in patients with late-onset depression and to correlate these abnormalities with clinical features. METHODS: The subjects, aged > or = 50 years, were 51 outpatients with major depression diagnosed according to DSM-IV criteria. They were living in the community and prospectively selected at their initial consultation for mostly mild-moderate depression. We also included 32 normal age-matched controls. We examined basic rhythms and temporal slow waves on EEG and determined the Hamilton Rating Scale for Depression (HAMD) as a scale for the severity of psychiatric symptoms. RESULTS: Temporal slow waves were more common in depressed patients (47%) than in normal controls (22%). Depressed patients with temporal slow waves had significantly lower frequency of family history of mood disorders (P<0.05). There was no difference in the total score of HAMD between patients with and without temporal slow waves, however, each score of HAMD in patients with temporal slow waves showed a significantly lower score for "feeling of guilt" (P<0.01) and a significantly higher score for "general somatic symptoms" (P<0.01) and "hypochondriasis" (P<0.01). CONCLUSIONS: Our EEG study suggested that temporal slow waves, which were probably associated with subtle cerebrovascular lesions, might reflect vulnerability to late-onset depression. In addition, depressed patients with temporal slow waves could be classified as a subgroup of heterogeneous late-onset depression based on the clinical findings. These results must be interpreted within the limitations of the sample selection procedure. PMID- 11869786 TI - Cryptosporidium parvum and Cyclospora cayetanensis: a review of laboratory methods for detection of these waterborne parasites. AB - Cryptosporidium and Cyclospora are obligate, intracellular, coccidian protozoan parasites that infest the gastrointestinal tract of humans and animals causing severe diarrhea illness. In this paper, we present an overview of the conventional and more novel techniques that are currently available to detect Cryptosporidium and Cyclospora in water. Conventional techniques and new immunological and genetic/molecular methods make it possible to assess the occurrence, prevalence, virulence (to a lesser extent), viability, levels, and sources of waterborne protozoa. Concentration, purification, and detection are the three key steps in all methods that have been approved for routine monitoring of waterborne oocysts. These steps have been optimized to such an extent that low levels of naturally occurring Cryptosporidium oocysts can be efficiently recovered from water. The filtration systems developed in the US and Europe trap oocysts more effectively and are part of the standard methodologies for environmental monitoring of Cryptosporidium oocysts in source and treated water. Purification techniques such as immunomagnetic separation and flow cytometry with fluorescent activated cell sorting impart high capture efficiency and selective separation of oocysts from sample debris. Monoclonal antibodies with higher avidity and specificity to oocysts in water concentrates have significantly improved the detection and enumeration steps. To date, PCR-based detection methods allow us to differentiate the human pathogenic Cryptosporidium parasites from those that do not infect humans, and to track the source of oocyst contamination in the environment. Cell culture techniques are now used to examine oocyst viability. While fewer studies have focused on Cyclospora cayetanensis, the parasite has been successfully detected in drinking water and wastewater using current methods to recover Cryptosporidium oocysts. More research is needed for monitoring of Cyclospora in the environment. Meanwhile, molecular methods (e.g. molecular markers such as intervening transcribed spacer regions), which can identify different genotypes of C. cayetanensis, show good promise for detection of this emerging coccidian parasite in water. PMID- 11869787 TI - Assessing bactericidal properties of materials: the case of metallic surfaces in contact with air. AB - A new method for assessing bactericidal properties of metallic materials, soiled by aerosol, was developed and applied to stainless steel in conditions close to reality. The airborne bacteria survival on different stainless steel grades and massive copper is presented here. The investigating bacterium was Enterococcus faecalis, which is a well-known contaminant strain in the indoor environments. It was observed that the bacterial aerosol lethality increased proportionally with the relative humidity (RH) of the environment. A significant difference in survival rate was measured depending on the tested supports, the greatest lethality being observed on clean massive copper. Moreover, the addition of nutrients on metallic surfaces, even in small quantities, was enough to ensure the revival of quiescent microorganisms. PMID- 11869788 TI - Enumeration of total bacteria and bacteria with genes for proteolytic activity in pure cultures and in environmental samples by quantitative PCR mediated amplification. AB - Real-time quantitative PCR assays were developed for the absolute quantification of different groups of bacteria in pure cultures and in environmental samples. 16S rRNA genes were used as markers for eubacteria, and genes for extracellular peptidases were used as markers for potentially proteolytic bacteria. For the designed 16S rDNA TaqMan assay, specificity of the designed primer-probe combination for eubacteria, a high amplification efficiency over a wide range of starting copy numbers and a high reproducibility is demonstrated. Cell concentrations of Bacillus cereus, B. subtilis and Pseudomonas fluorescens in liquid culture were monitored by TaqMan-PCR using the 16S rDNA target sequence of Escherichia coli as external standard for quantification. Results agree with plate counts and microscopic counts of DAPI stained cells. The significance of 16S rRNA operon multiplicity to the quantification of bacteria is discussed.Furthermore, three sets of primer pair together with probe previously designed for targeting different classes of bacterial extracellular peptidases were tested for their suitability for TaqMan-PCR based quantification of proteolytic bacteria. Since high degeneracy of the probes did not allow accurate quantification, SybrGreen was used instead of molecular probes to visualize and quantify PCR products during PCR. The correlation between fluorescence and starting copy number was of the same high quality as for the 16S rDNA TaqMan assay for all the three peptidase gene classes. The detected amount of genes for neutral metallopeptidase of B. cereus, for subtilisin of B. subtilis and for alkaline metallopeptidase of P. fluorescens corresponded exactly to the numbers of bacteria investigated by the 16S rDNA targeting assay. The developed assays were applied for the quantification of bacteria in soil samples. PMID- 11869789 TI - The effect of reducing agents on the recovery of injured Bifidobacterium cells. AB - Bifidobacterium spp. have been proposed as a microbial indicator of faecal pollution in water and as possible probiotic in fermented dairy products. These anaerobic bacteria can be subject to metabolic stress in environmental samples because of the presence of oxygen or its derivatives. In this case, their recovery is compromised, especially in selective media. Three reducing agents were tested to improve the recovery of oxygen-stressed bifidobacteria: L cysteine, sodium pyruvate and sodium thioglycolate. These agents were evaluated individually at various concentrations, and in combination by measuring recovery on rich and selective media. The addition of several mixtures of reducing agents to the water samples and pre-incubation for 4 h at 37 degrees C increased the recovery of Bifidobacterium spp. on culture media. PMID- 11869790 TI - Evaluation of extraction and purification methods for obtaining PCR-amplifiable DNA from compost for microbial community analysis. AB - Analysis of microbial community structure in complex environmental samples using nucleic acid techniques requires efficient unbiased DNA extraction procedures; however, humic acids and other contaminants complicate the isolation of PCR amplifiable DNA from compost and other organic-rich samples. In this study, combinations of DNA extraction and purification methods were compared based on DNA yield, humic acid contamination, PCR amplifiability, and microbial community structure assessed by terminal restriction fragment length polymorphisms (TRFLP) of amplified 16S rRNA genes. DNA yield and humic acid contamination, determined by A230, varied significantly between extraction methods. Humic acid contamination of DNA obtained from compost decreased with increasing salt concentration in the lysis buffer. DNA purified by gel permeation chromatography (Sepharose 4B columns) gave satisfactory PCR amplification with universal eubacterial 16S rRNA gene primers only when A260/A280 ratios exceeded 1.5. DNA purified with affinity chromatography (hydroxyapatite columns), and showing A260/A280 ratios as high as 1.8, did not show consistently satisfactory PCR amplification using the same 16S rRNA primers. Almost all DNA samples purified by agarose gel electrophoresis showed satisfactory PCR amplification. Principal components analysis (PCA) of TRFLP patterns differentiated compost types based on the presence/absence of peaks and on the height of the peaks, but differences in TRFLP patterns were not appreciable between extraction methods that yielded relatively pure DNA. High levels of humic acid contamination in extracted DNA resulted in TRFLP patterns that were not consistent and introduced a bias towards lower estimates of diversity. PMID- 11869791 TI - A promoter-trap vector for clock-controlled genes in the cyanobacterium Synechocystis sp. PCC 6803. AB - We constructed a promoter-trap vector pPT6803-1 to isolate circadian clock controlled promoters in the cyanobacterium Synechocystis sp. strain PCC 6803. The vector contains a promoterless luciferase gene set (luxAB) from Vibrio harveyi that is targeted to a specific site of the Synechocystis genome as a reporter for gene expression. A library was constructed in pPT6803-1 by introducing the genomic DNA fragments upstream of luxAB to transform Synechocystis cells. Of approximately 10,000 Synechocystis transformants, at least 55 (#1-55) showed circadian rhythms of bioluminescence under continuous illumination. Clones #19, #22, and #26 exhibited obviously different waveforms of bioluminescence from each other. Deletion analysis and primer extension experiments mapped the promoters for the clpP, slr1634, and rbpP genes that are responsible for bioluminescence from #19, #22, and #26, respectively. PMID- 11869792 TI - Bacterial genomics: the use of DNA microarrays and bacterial artificial chromosomes. AB - Immense amounts of genetic information are contained within microbial genomes. As the number of completely sequenced microbial genomes is increasing, functional and comparative genomic techniques will be employed for sequence analysis and gene characterization. Sequence comparison and expression profiling by DNA microarrays can determine phylogenetic relationships and identify genes while bacterial artificial chromosomes (BACs) allow the study of entire biochemical pathways and permit the expression of bacterial genes in a foreign host. PMID- 11869793 TI - Bactericidal activity of electrolyzed acid water from solution containing sodium chloride at low concentration, in comparison with that at high concentration. AB - Electrolyzed strong acid water (ESW) containing free chlorine at various concentrations is becoming to be available in clinical settings as a disinfectant. ESW is prepared by electrolysis of a NaCl solution, and has a corrosive activity against medical instruments. Although lower concentrations of NaCl and free chlorine are desired to eliminate corrosion, the germicidal effect of ESW with low NaCl and free-chlorine concentrations (ESW-L) has not been fully clarified. In this study, we demonstrated that ESW-L possesses bactericidal activity against Mycobacteria and spores of Bacillus subtilis. The effect was slightly weaker than that of ESW containing higher NaCl and free-chlorine concentrations (ESW-H), but acceptable as a disinfectant. To clarify the mechanism of the bactericidal activity, we investigated ESW-L-treated Pseudomonas aeruginosa by transmission electron microscopy, a bacterial enzyme assay and restriction fragment length polymorphism pattern (RFLP) assay. Since the bacterium, whose growth was completely inhibited by ESW-L, revealed the inactivation of cytoplasmic enzyme, blebs and breaks in its outer membrane and remained complete RFLP of DNA, damage of the outer membrane and inactivation of cytoplasmic enzyme are the important determinants of the bactericidal activity. PMID- 11869794 TI - Reliability of hsp65-RFLP analysis for identification of Mycobacterium species in cultured strains and clinical specimens. AB - The purpose of this study was to evaluate the reliability of an amplification restriction analysis based method (hsp65-RFLP) to detect and identify mycobacterial species in clinical samples and cultures with low number of bacilli. We examined 247 clinical specimens and 88 culture vials, comparing hsp65 RFLP results with conventional culture/biochemical tests. The analytical sensitivity of the method was assessed with cerebrospinal fluid (CSF), broncho alveolar lavage (BAL), sputum, water, and 12B medium containing defined amounts of mycobacterial chromosome. We detected the equivalent of 10(3) cells per ml in all samples, except sputum, the most common source of clinical sample for mycobacterial testing, which presented inhibition throughout. We investigated two purification procedures to overcome inhibition of DNA amplification: DNAzol and phenol/chloroform. The former was superior, eliminating inhibition in 93.7% of the clinical samples. The technique was effective for bacterial cultures, including those with very low growth indices (GIs), substantially abbreviating time for diagnosis, but showed low sensitivity (25%) when applied to clinical samples, an issue that has never been extensively assessed by other researchers. PMID- 11869796 TI - Automated measurements of antilisterial activities of lactate and diacetate in ready-to-eat meat. AB - Standard procedures to enumerate Listeria organisms rely on plating food samples on selective agar media. The procedures are labor-intensive, and because of the limited sensitivity, pre-enrichment step is required for the detection of low numbers of the pathogen. In the present study, an automated rapid optic procedure and the standard procedure were used to determine the behavior of the pathogen in ready-to-eat (RTE) meat and to test the effect of antilisterial agents. Listeria monocytogenes strain Scott A or a six-strain mixture of Listeria was studied using lactate (2.5%), diacetate (0.2%) and their combination in beef bologna and in sterile beef emulsion. Samples stored for up to 60 days at 5 and 10 degrees C were tested at time intervals during storage. Using the plate count method, each of the salts caused a delay in growth of the pathogen, and the salt combination was most effective causing listeriostatic effects and decline in growth of the pathogen at 5 degrees C. High negative correlation (r), ranging from 0.92 to 0.99, was obtained between the detection time (DT) recorded by the optic procedure (BioSys instrument) and cell numbers determined by the plate count procedure. The rapid (< 24 h) optic procedure was reliable in assessing the efficacy of antimicrobials and in rapid detection of low levels of listeriae that are undetectable by direct plating procedure. PMID- 11869795 TI - Microbial identification by immunohybridization assay of artificial RNA labels. AB - Ribosomal RNA (rRNA) and engineered stable artificial RNAs (aRNAs) are frequently used to monitor bacteria in complex ecosystems. In this work, we describe a solid phase immunocapture hybridization assay that can be used with low molecular weight RNA targets. A biotinylated DNA probe is efficiently hybridized in solution with the target RNA, and the DNA-RNA hybrids are captured on streptavidin-coated plates and quantified using a DNA-RNA heteroduplex-specific antibody conjugated to alkaline phosphatase. The assay was shown to be specific for both 5S rRNA and low molecular weight (LMW) artificial RNAs and highly sensitive, allowing detection of as little as 5.2 ng (0.15 pmol) in the case of 5S rRNA. Target RNAs were readily detected even in the presence of excess nontarget RNA. Detection using DNA probes as small as 17 bases targeting a repetitive artificial RNA sequence in an engineered RNA was more efficient than the detection of a unique sequence. PMID- 11869797 TI - Long PCR-amplified rDNA for PCR-RFLP- and Rep-PCR-based approaches to recognize closely related microbial species. AB - Long PCR was used to amplify a 5-kb fragment of the bacterial ribosomal operon (16S-intergenic spacer region (ISR)-23S) from several Ralstonia eutropha strains (16S rDNA sequence similarity: 97-99%). Due to the large product size, amplicons from the different strains could be distinguished using restriction enzyme fragment length polymorphisms (RFLP) and repetitive PCR analysis (Rep-PCR) with the primer 1492r. These methods may prove useful in differentiating other bacterial strains with highly similar 16S rDNA sequences. PMID- 11869798 TI - A simple and rapid assay of collagen-like polymer in crude lysate from Escherichia coli. AB - An assay for the quantification of collagen-like polymer (CLP) in Escherichia coli cells utilizing the specific reaction between collagenase and CLP is presented. It involves thermal treatment to precipitate non-CLP proteins, digestion of CLP by collagenase and detection of the absorbance of the liberated amino acids and peptides from CLP by a ninhydrin-based method. CLP concentration is determined from the absorbance measurement. PMID- 11869799 TI - Potassium release, a useful tool for studying antimicrobial peptides. AB - We used an ion-selective electrode to measure potassium release from bacteria treated with antimicrobial peptides. This broadly applicable and simple technique for assessing membrane integrity deserves greater use. PMID- 11869800 TI - Development of a method for heterologous gene expression in Enterobacter amnigenus, a potential host for the biological control of mosquito larvi. AB - An integrative plasmid containing a 1.3 kb fragment of chromosomal DNA from Enterobacter amnigenus was constructed. The Omega fragment encoding spectinomycin/streptomycin resistance was cloned into the unique BglII site of the resulting plasmid, and the interrupted fragment was transferred via plasmid pMAK705 by electroporation into E. amnigenus with a selection for spectinomycin resistance. Cointegrants were resolved to generate an E. amnigenus strain that expressed spectinomycin resistance, but grew as rapidly as the parental strain. The cloned fragment encodes a putative homologue of the proW gene of Escherichia coli that is not essential for E. amnigenus growth. The integrative plasmid is now available to introduce any heterologous DNA into the E. amnigenus chromosome, for the construction of promoter-probe vectors for the studies of gene regulation, or to construct plasmids suitable for the isolation of secretion signals. Immediate applications of this system will include the expression and secretion of crystal toxins from bacilli for the biological control of mosquito larvae infected with the bacterial host. PMID- 11869801 TI - Immuno-capture PCR for detection of Aeromonas hydrophila. AB - In this report, we describe the use of universal primer PCR (UPPCR) for the detection of 16S ribosomal RNA (rRNA) genes from Aeromonas hydrophila captured by anti-A. hydrophila antibody coupled to a microplate. The approach combining immuno-capture with UPPCR provides a quick, sensitive, and reproducible way for the detection of bacterial cells. PMID- 11869802 TI - Cellular expression and subcellular localization of the human Ins(1,3,4,5)P(4) binding protein, p42(IP4), in human brain and in neuronal cells. AB - In this study we describe for the human inositol-(1,3,4,5)-tetrakisphosphate (InsP4)-binding protein, p42IP4, the cellular distribution and subcellular localization in human brain and in transfected neuronal cells. The cDNA sequence of the human p42IP4 containing a single open reading frame yields a peptide of 374 amino acids with a calculated molecular mass of 43.4 kDa with a zinc-finger motif at the N-terminus, followed by two pleckstrin homology (PH) domains. Using a peptide-specific antiserum, p42IP4 protein was localized in a majority of neuronal cells of human brain sections. In the hypothalamus a subpopulation of paraventricular and infundibular nucleus neurons were strongly immunoreactive for p42IP4. In cortical areas the protein was predominantly found in large pyramidal cells. Some immunoreactivity for p42IP4 was also observed in the pyramidal cells of the hippocampal formation. Functional expression of p42IP4 protein in neuronal (NG108-15) and non-neuronal (CHO-K1) cells stably transfected with GFP-p42IP4 was shown in all cell fractions (homogenate, cytosol and membranes) by specific [3H]Ins(1,3,4,5)P4 binding activity, which correlated with p42IP4 protein detection by Western blot analysis. Using confocal laser scanning microscopy we showed that in NG108-15 and CHO-K1 cells stably transfected with GFP-p42IP4 the full length p42IP4 protein was localized in the cytoplasm, at the plasma membrane and in the nucleus. A deletion mutant of p42IP4 lacking the zinc finger domain resulted in solely a cytosolic and membrane localization but was not found in the nucleus. Thus we can conclude that human p42IP4 shows a region-specific localization in the human brain and the zinc finger motif within the protein is responsible for the localization of the protein in the cell nucleus. PMID- 11869803 TI - Effects of ferroxidase activity and species on ceruloplasmin mediated iron uptake by BT325 cells. AB - In a previous study, we found that human ceruloplasmin (hCP) promotes iron uptake rather than release in BT325 cells, a human glioma cell line. In this study, we investigated the effect of ferroxidase activity of hCP and different species of ceruloplasmins on CP-mediated iron uptake by the cells. The cells were incubated for 30 min at 37 degrees C with 1 microM 59Fe2+ with or without 150 microg/ml of the untreated and the ferroxidase-defective hCPs (apohCP and heat-inactivated hCP) or different species of ceruloplasmins (human CP, rabbit CP and bovine CP). The untreated hCP induced a significant increase in iron uptake by BT325 cells, while ferroxidase-defective hCPs with (heat-inactivated hCP) or without cooper ions (apohCP) had no such role. The untreated hCP increases significantly internalized iron but not membrane-bound iron, implying that hCP stimulated iron entry into the cell rather than increased extracellular binding of iron to the cell surface. All species of ceruloplasmins could promote iron uptake by the cells and the difference in degree of stimulatory effect among them was insignificant. These results suggested that ferroxidase activity of hCP is essential for the hCP-mediated iron uptake process and also that CP-stimulated iron uptake is not associated with copper ions in the protein, and that the effect of ceruloplasmin on iron uptake by BT325 cells is not species specific. PMID- 11869804 TI - Expression and activity of nitric oxide synthase isoforms in rat brain during the development of experimental allergic encephalomyelitis. AB - The activity and expression of nitric oxide synthase (NOS) isoforms and protein nitrotyrosine (NT) residues were investigated in whole encephalic mass (WEM) homogenates during the development of experimental allergic encephalomyelitis (EAE) in Lewis rats. EAE stages (0-III) were daily defined by clinical evaluation, and in the end of each stage, WEMs were removed for analysis of NOS activity, protein NT residues and mRNA for the different NOS isoforms. In the presence of NADPH, WEMs from EAE-III rats showed lower Ca2+-dependent NOS activity than those from control group. These differences disappeared in the presence of exogenous calmodulin, flavin adenine dinucleotide (FAD), tetrahydrobiopterin (BH4) and NADPH. Of all the cofactors, just the omission of FAD caused comparable decrease of Ca2+-dependent NOS activity from both groups. Ca2+-independent NOS activity from EAE-III animals was insensitive to the omission of any of the cofactors, while in control animals this activity was significantly inhibited by the omission of either FAD or BH4. Increased levels of both iNOS mRNA and protein NT expression were observed in animals with EAE, which also showed lower levels of a thermolabile NOS inhibitor in WEM homogenates and sera than controls. In conclusion, during late EAE stages, constitutive Ca2+ dependent NOS activity decreases concomitantly with iNOS upregulation, which could be responsible for the high protein NT levels. The differential dependence of iNOS activity on cofactors and the absence of an endogenous thermolabile NOS inhibitor in animals with EAE could reflect additional control mechanisms of NOS activity in this model of multiple sclerosis. PMID- 11869805 TI - Cold stress alters Mytilus edulis pedal ganglia expression of mu opiate receptor transcripts determined by real-time RT-PCR and morphine levels. AB - Previous pharmacological, biochemical and molecular evidence prove that mu subtype opiate receptors and opiate alkaloids, i.e. morphine, are present in the ganglionic nervous system of the mollusk Mytilus edulis (bivalve). We now present molecular evidence on the effect of rapid temperature changes on mu opiate receptor expression and morphine levels. Using primers, a labeled Taq-Man probe derived from the human neuronal mu1 opiate receptor, and real-time RT-PCR to measure the expression of mu transcripts from Mytilus pedal ganglia, we observe, in animals placed in cold water from room temperature, an enhanced morphine and morphine 6 glucuronide level in addition to a decrease in mu opiate receptor gene expression. This study provides further evidence that mu-type opiate receptors and morphine are expressed in mollusk ganglia and appear to be involved in physiological processes responding to thermal stress. PMID- 11869806 TI - Retinoic acid regulation of mu opioid receptor and c-fos mRNAs and AP-1 DNA binding in SH-SY5Y neuroblastoma cells. AB - Retinoic acid (RA) induced differentiation of SH-SY5Y cells increases the expression of mu opioid receptors (HMOR) and inhibitory G proteins, as well as the efficacy of opioids to inhibit forskolin-induced adenylyl cyclase activity. We examined the time course of the effects of all-trans retinoic acid (RA) on HMOR and c-fos mRNA levels as determined by solution hybridization (using HMOR and rat c-fos riboprobes) in RNA extracts from SH-SY5Y cells. Electrophoretic Mobility Shift Assay (EMSA) and Western blot analysis were used to assess the changes AP-1 DNA binding and the presence of fos-related proteins in nuclear extracts from untreated, vehicle (ethanol) or RA-treated SH-SY5Y cells. Exposure to RA for 0.5 h had no effect on HMOR while after 6-18 h of exposure HMOR in mRNA levels were decreased by 50% and then after 168 h of RA exposure, HMOR mRNA levels were doubled. In contrast, c-fos mRNA levels were unchanged at 0.5 h, but increased by 50% after 18 and 168 h of RA exposure. RA increased AP-1 binding after 18 and 168 h and a pan fos-FRA antibody produced a supershift. Western analysis indicates that RA activates a 45-kDa protein corresponding to the size of the fos B protein. These results identify two signal transduction targets that are regulated by RA during differentiation. PMID- 11869807 TI - Age-related changes in the protein expression of subunits of the NMDA receptor. AB - C57Bl/6 mice show decreased expression of the mRNA for the epsilon2 and zeta1 subunits of the N-methyl-D-aspartate (NMDA) receptor in subregions of the cerebral cortex and hippocampus with increased age. The purpose of this study was to determine the effects of aging on the protein expression of the three major subunits of the NMDA receptor. Semi-quantitative Western blot techniques were applied with the use of antibodies that recognize either the epsilon1 (NR2A), epsilon2 (NR2B) or zeta1 (NR1) subunits of the NMDA receptor or a synaptic terminal protein, synaptophysin. In the cerebral cortex of 30-month-old mice, the level of protein expression of both the epsilon2 and zeta1 subunits were decreased significantly from levels found in the 3- and 10-month-old mice and the protein expression of the epsilon1 subunit showed a significant decline between 10 and 30 months of age. In the hippocampus, the epsilon2 subunit exhibited a higher protein expression level in the 10-month-old mice as compared to both the young and old mice and the zeta1 subunit showed a significant drop in expression in the old mice from both 3- and 10-month-olds. Synaptophysin showed significant declines in protein expression with increasing age. These results demonstrated that changes in the protein expression of the major subunits of the NMDA receptor occur during the aging process and, in some cases, were greater than changes seen previously in mRNA expression. These subunit alterations may explain some of the changes that are seen in NMDA receptor functions during aging. PMID- 11869808 TI - Abundant type 10 17 beta-hydroxysteroid dehydrogenase in the hippocampus of mouse Alzheimer's disease model. AB - A full-length cDNA of mouse type 10 17 beta-hydroxysteroid dehydrogenase (17 beta HSD10) was cloned from brain, representing the accurate nucleotide sequence information that rendered possible an accurate deduction of the amino acid sequence of the wild-type enzyme. A comparison of sequences and three-dimensional models of this enzyme revealed that structures previously reported by other groups carry either a truncated or mutated amino-terminal sequence. Fusion of the first 11 residues of the wild-type enzyme to the green fluorescent protein directed the reporter protein into mitochondria. Thus, the N-terminus was identified as a mitochondrial targeting signal that accounts for the intracellular localization of the mouse enzyme. This enzyme is normally associated with mitochondria, not with the endoplasmic reticulum as suggested by its trivial name 'endoplasmic reticulum-associated amyloid-beta biding protein (ERAB)'. After its C-terminal region was used to raise rabbit anti-17 betaHSD10 antibodies, immunogold electron microscopy showed that an abundance of this enzyme could be found in hippocampal synaptic mitochondria of betaAPP transgenic mice, but not in normal controls. High levels of this enzyme may disrupt steroid hormone homeostasis in synapses and contribute to synapse loss in the hippocampus of the mouse Alzheimer's disease model. PMID- 11869809 TI - Internalization of metabotropic glutamate receptor in C6 cells through clathrin coated vesicles. AB - C6 glioma cells were treated for hours with 100 microM L-glutamate, quisqualate or trans-ACPD. In all cases, phospholipase C-coupled metabotropic glutamate receptors (mGluRs) present in these cells are down-regulated after this agonist treatment. Cell surface metabotropic glutamate receptor density was minimum at 6 h of agonist treatment and reached near control values after 30 h of treatment. This recovery was associated with a progressive increase in mGluR1 and mGluR1a mRNA level between 6 and 24 h and was not due to agonist removal. Specific L [3H]glutamate or [3H](+/-)trans-ACPD binding decrease detected in C6 cells after 6 h of 100 microM L-glutamate treatment was associated with a remarkable increase of specific L-[3H]glutamate binding detected in clathrin-coated vesicles isolated from these treated cells. Moreover, this decrease was blocked in the presence of 0.5 M sucrose or 1 microM phenylarsine oxide, suggesting that desensitization and down-regulation of mGluR can be due to an endocytosis process through clathrin coated pits and vesicles. PMID- 11869810 TI - Rapid deafferentation-induced upregulation of bcl-2 mRNA in the chick cochlear nucleus. AB - Neuronal survival in developing animals is often dependent on afferent activity. In the posthatch chick, approximately 30% of the neurons in the avian cochlear nucleus, nucleus magnocellularis (NM) die following elimination of VIIIth nerve activity. The factors that influence death or survival of an individual NM neuron are largely unknown. Previous studies indicate that both cell death and cell survival mechanisms compete to determine cell fate. One factor that has been shown to suppress cell death cascades in a variety of systems is bcl-2. If this gene product plays a role in regulating cell survival following deafferentation, then one might expect the expression of this gene to be influenced by removal of afferent input. In the present study, in situ hybridization revealed a rapid and transient increase in bcl-2 mRNA in NM neurons following deafferentation. Enhanced bcl-2 mRNA expression was observed at 6 and 12 h following deafferentation, but not at 3 or 24 h. Surprisingly, the upregulation of bcl-2 mRNA was limited to a subpopulation (20-30%) of deafferented neurons corresponding to the number of NM neurons that eventually die following cochlea removal. The robust and rapid upregulation of this gene suggests that cell death cascades regulated by bcl-2 may be initiated following deafferentation. PMID- 11869811 TI - Cortical spreading depression transiently activates MAP kinases. AB - Cortical spreading depression (CSD) has been shown to have neuroprotective effects when administered in advance of cerebral ischemia. The mechanism by which CSD induces its neuroprotective effect however remains to be elucidated. Since MAP kinases have been shown to impart neuroprotection in ischemic preconditioning paradigms, we attempted to determine the role CSD may have in the activation of MAPK. We show that CSD is capable of increasing the phosphorylation of ERK in a MEK-dependent manner. This phosphorylation is, however, transient, as phosphorylated ERK levels return to control levels 45 min after 2 h of CSD elicitation. Immunohistochemical analysis reveals that the phosphorylated form of ERK is located ubiquitously in cells of the CSD-treated cortex while CSD-elicited MEK phosphorylation resides solely in the nuclei. These data suggest that CSD may act via the MAP kinase pathways to mediate preconditioning. PMID- 11869813 TI - Global harmonisation of classification and labelling of hazardous chemicals. AB - The Globally Harmonised System (GHS) for classification and labelling will provide an internationally agreed system for hazard classification of chemical products and for communication of those hazards. Under the system, chemicals will be classified according to their physical (e.g. flammability), health/toxicological and environmental hazards. The toxicological endpoints used in the system are acute toxicity, irritation or corrosivity, sensitisation, carcinogenicity, mutagenicity, reproductive toxicity and chronic or repeat dose toxicity. The intended target audiences are those concerned with transport of dangerous goods, consumers, workers and emergency responders. Labels and safety data sheets (SDS) are the core tools of the GHS hazard communication system, and the harmonised labelling elements are symbols (within a pictogram), signal words and hazard statements. The GHS will use a building block approach in which application may vary according to the circumstances, type of product, and stage of life cycle, allowing selection of the elements appropriate to the needs of the various end users. PMID- 11869814 TI - Unexpected tumour findings in lifetime rodent bioassay studies--what to do? AB - Currently, the majority of substances tested in lifetime bioassays in rodents are not mutagenic and, therefore, at the most weakly carcinogenic, generally by epigenetic mechanisms. It thus appears obvious that only marginal increases of tumour incidences can be expected in lifetime bioassays and that, therefore, every aspect of a potential carcinogenic effect must be thoroughly evaluated. This paper describes a series of key factors, which should be looked at in order to exclude that the lifetime bioassay in question is flawed for design, technical or qualification reasons. It also provides some hints whether there is indeed a real effect and not just a variation of the spontaneous tumour incidences. Tumour findings must be seen in the context of the animal model, the pharmcokinetics and pharmcodynamics of the test substance, as well as any other observation in the present or other studies with the test substance, including non-tumour findings and--in particular--potential precursor lesions and effects on feed intake and survival. The possibility that the observed carcinogenic effects may be species specific and not relevant for man is discussed. It is also important to check what findings are reported with similar substances or substances with the same pharmacological effect. Data from additional investigations on material of the same study and/or mechanistic studies are often needed to support the final risk assessment. PMID- 11869815 TI - The EU precautionary bans of animal feed additive antibiotics. AB - Toxicologists, with good reason, will feel that the biological safety of chemical products across the market sectors rests largely on their efforts. However, one sector has received much adverse attention from the media, consumers, politicians, legislators and advisory groups in recent years. It is food animal production in intensive systems and, within those, various types of chemical additives included in the compound diets fed. No additive class received more adverse comment than those antibiotics used for the purpose of enhancing the efficiency of animal production. This paper considers the safety of the antibiotic feed additives (AFAs) against the background of the regulatory measures in place, defines their role and describes the relevant concerns. It closes with comment on the microbiologically-based health risk which underpinned the AFA bans and sounds a warning over the precedent created by the use of the precautionary principle in the recent banning of six of their number. PMID- 11869816 TI - Environmental risk assessment of pesticides on aquatic life in Xiamen, China. AB - This paper describes the results of the environmental risk assessment of the pesticides used in agriculture in Xiamen, China. The goal was to assess the impact on water resources, particularly on fisheries and mariculture. Data on ecotoxicological properties of the pesticides and their physico-chemical profile were collected. The simulation of the environmental behaviour of the pesticides in relation to the load applied onto the agricultural areas was done using the SoilFug model. Risk assessment was performed, pesticides approximate concentrations have been calculated, chemicals at highest risk were identified, and risk management measures were indicated. This study could represent a cost effective method that may be used before engaging in expensive monitoring programs for pesticide use in developing countries, where analytical facilities are lacking. PMID- 11869817 TI - The wreckage of the oil tanker 'Erika'--human health risk assessment of beach cleaning, sunbathing and swimming. AB - In December 1999 the oil tanker 'Erika', carrying approximately 30 tons of heavy fuel oil, wrecked before the coast of Brittany (France), polluting the local beaches and rocks over a distance of some 500 km. Also numerous birds were affected. During the first months of 2000 the coastal area and many birds were cleaned. The health risk for people involved in these cleaning activities and for tourists was evaluated with emphasis on the carcinogenic properties of this oil. The outcome indicates that the risks were limited to people who had been in bare handed contact with the oil. Firstly they had an increased risk for developing skin irritation and dermatitis, however, these effects are in general reversible. Secondly they had an increased risk for developing skin tumours, but since the dermal contacts with the oil were of relative short duration, this risk is considered to be very limited. PMID- 11869818 TI - Chronic arsenic poisoning. AB - Symptomatic arsenic poisoning is not often seen in occupational exposure settings. Attempted homicide and deliberate long-term poisoning have resulted in chronic toxicity. Skin pigmentation changes, palmar and plantar hyperkeratoses, gastrointestinal symptoms, anemia, and liver disease are common. Noncirrhotic portal hypertension with bleeding esophageal varices, splenomegaly, and hypersplenism may occur. A metallic taste, gastrointestinal disturbances, and Mee's lines may be seen. Bone marrow depression is common. 'Blackfoot disease' has been associated with arsenic-contaminated drinking water in Taiwan; Raynaud's phenomenon and acrocyanosis also may occur. Large numbers of persons in areas of India, Pakistan, and several other countries have been chronically poisoned from naturally occurring arsenic in ground water. Toxic delirium and encephalopathy can be present. CCA-treated wood (chromated copper arsenate) is not a health risk unless burned in fireplaces or woodstoves. Peripheral neuropathy may also occur. Workplace exposure or chronic ingestion of arsenic-contaminated water or arsenical medications is associated with development of skin, lung, and other cancers. Treatment may incklude the use of chelating agents such as dimercaprol (BAL), dimercaptosuccinic acid (DMSA), and dimercaptopanesulfonic acid (DMPS). PMID- 11869819 TI - Arsenic induced changes in growth development and apoptosis in neonatal and adult brain cells in vivo and in tissue culture. AB - Arsenic at a nonlethal level in drinking water consumed over a period of time has been reported to produce chronic toxicity and various types of health problems ranging from skin cancer to disturbance in memory. Neurotoxic effects have been reported in clinical cases with chronic exposure to arsenic. Physiological detoxication of arsenic occurs partially through methylation. Arsenic and its methylated derivatives are distributed in different organs and systems. The present study examined the possible interference in the neuronal development and differentiation due to the exposure to arsenic during gestation. The experiments were carried out to examine short and long term effects of arsenic on brain explants and cells grown and maintained in tissue culture system. The effects of arsenic exposure showed changes in brain cell membrane function indicated by generation and release of reactive oxygen-nitrogen intermediates. On the morphological aspect the explants' growth was reduced, ground matrix was lost and neural networking was inhibited. Cells showed signs of apoptotic changes. Arsenic toxicity may induce damage to brain cells prior to more visible clinical conditions. The deleterious effects also pass from the maternal to fetal tissue across the transplacental barrier. PMID- 11869820 TI - Mechanisms of multiple chemical sensitivity. AB - Sensitivity to chemicals is a toxicological concept, contained in the dose response relationship. Sensitivity also includes the concept of hypersensitivity, although controversy surrounds the nature of effects from very low exposures. The term multiple chemical sensitivity has been used to describe individuals with a debilitating, multi-organ sensitivity following chemical exposures. Many aspects of this condition extend the nature of sensitivity to low levels of exposure to chemicals, and is a designation with medical, immunological, neuropsychological and toxicological perspectives. The basis of MCS is still to be identified, although a large number of hypersensitivity, immunological, psychological, neurological and toxicological mechanisms have been suggested, including: allergy; autosuggestion; cacosomia; conditioned response; immunological; impairment of biochemical pathways involved in energy production; impairment of neurochemical pathways; illness belief system; limbic kindling; olfactory threshold sensitivity; panic disorder; psychosomatic condition; malingering; neurogenic inflammation; overload of biotransformation pathways (also linked with free radical production); psychological or psychiatric illness; airway reactivity; sensitisation of the neurological system; time dependent sensitisation, toxicant induced loss of tolerance. Most of these theories tend to break down into concepts involving: (1) disruption in immunological/allergy processes; (2) alteration in nervous system function; (3) changes in biochemical or biotransformation capacity; (4) changes in psychological/neurobehavioural function. Research into the possible mechanisms of MCS is far from complete. However, a number of promising avenues of investigation indicate that the possibility of alteration of the sensitivity of nervous system cells (neurogenic inflammation, limbic kindling, cacosomia, neurogenic switching) are a possible mechanism for MCS. PMID- 11869821 TI - Is multiple chemical sensitivity a clinically defined entity? AB - In 1996 a WHO/IPCS Workshop has suggested to use as an appropriate descriptor of MCS the broader term "Idiopathic Environmental Intolerances (IEI)", in order to incorporate "a number of disorders sharing similar symptomatologies". Research was strongly encouraged. The following points have been put forward as a precondition to define MCS as a clinical entity: (a) establishment of diagnostic criteria, (b) identification of pathogenic mechanisms, together with, (c) an explanation of relationship between exposures and symptoms. Against this background, progress made in the fields of sensory physiology and neurobehaviour research must be debated. In particular, recent results on processing of cognitive stimuli have to be considered. IEI/MCS patients exhibited differences vs. controls in their reactions to intranasal challenge, consistent with changes in cognitive processing of suprathreshold chemosensory information. Trait anxiety and focus of attention have clearly been identified as major components in eliciting neurobehavioural MCS symptoms. Hence, the question as to whether MCS should be regarded as a clinically defined entity remains controversial, but important progress can be noticed in elucidating and defining the nature of this phenomenon, by a combined effort of several disciplines (toxicology and behavioural toxicology, psychology and psychophysiology, and clinical medicine). The new situation will call for a re-evaluation of traditional positions. PMID- 11869822 TI - Early spatial memory deficit induced by 2,5-hexanedione in the rat. AB - 2,5-Hexanedione (2,5-HD), the major common neurotoxic metabolite of n-hexane and methyl n-butyl ketone, causes a delayed neuropathy with associated sensorimotor impairments. The question arises as to whether specific cognitive deficits occur even prior to changes in sensorimotor ability. The present experiments examined the effects of 2,5-HD on spatial navigation of rats in a water maze at levels/times that did not affect spontaneous exploratory motor activity in an open field holeboard apparatus. Exposure to 1% 2,5-HD in the drinking water for 2 weeks did not significantly affect escape learning, as measured by latency to find a hidden platform. However, 2,5-HD treated animals were impaired in the use of a spatial strategy during a recall test. A similar impairment in spatial memory was observed after i.p. injection of 500 mg/kg/day 2,5-HD for 4 days, in the absence of significant changes in sensorimotor ability or weight loss. Thus 2,5-HD may mediate some of the cognitive effects of hexacarbons and these changes can occur prior to the development of motor symptoms. PMID- 11869823 TI - Extrathyroidal actions of antithyroid thionamides. AB - Some compounds having thionamide structure inhibit thyroid functions. Such antithyroid thionamides include mercaptomethylimidazole (methimazole), thiourea and propylthiouracil, of which mercaptomethylimidazole is widely used to treat hyperthyroidism. Undesirable side effects develop from these drugs due to extrathyroidal actions. Antithyroid thionamides inhibit lactoperoxidase which contributes to the antibacterial activities of a number of mammalian exocrine gland secretions that protect a variety of mucosal surfaces. These drugs stimulate both gastric acid and pepsinogen secretions, thereby augmenting the severity of gastric ulcers and preventing wound healing. Increased gastric acid secretion is partially due to the H2 receptor activation, and also through the stimulation of the parietal cell by intracellular generation of H2O2 following inactivation of the gastric peroxidase-catalase system. Severe abnormalities may develop in blood cells and the immune system after thionamide therapy. It causes agranulocytosis, aplastic anemia, and purpura along with immune suppression. Olfactory and auditory systems are also affected by these drugs. Thionamide affects the sense of smell and taste and also causes loss of hearing. It binds to the Bowman's glands in the olfactory mucosa and causes extensive lesion in the olfactory mucosa. Thionamides also affect gene expression and modulate the functions of some cell types. A brief account of the chemistry and metabolism of antithyroid thionamides, along with their biological actions are presented. PMID- 11869825 TI - Estrogenic isoflavones in rodent diets. AB - Many rodent diets contain components such as soy isoflavones (daidzein and genistein) known to have estrogenic properties. The dietary background of phytoestrogens may modulate some responses to environmental estrogens when these compounds are tested in rodent bioassays. Thus, and since only few data were available on the phytoestrogen content of rodent diets commonly used in European laboratories, it was of interest to analyze the daidzein and genistein contents of our standard animal feeds. Isoflavone contents were determined in seven batches of rodent chow (from two suppliers in Germany, Altromin and Ssniff) by high-performance liquid chromatography, and also analyzed in six rodent diets from the United States. The soy-based rodent diets from Germany contained isoflavone (daidzein plus genistein) concentrations in the range of 0.3-0.55 mg/g feed. These isoflavone contents are similar to those analyzed in the US rodent diets, and similar to values reported by others, including one particular lot of feed (with 0.35 mg isoflavones per g) which produced a large uterotrophic response in immature ovariectomized rats [Environ. Health Perspect., 106 (1998) 369]. Coumestrol was found in a sample of commercial rabbit food at rather high levels (0.27 mg/g), but, this phytoestrogen was not detected (<1 microg/g feed) in any of the other samples we analyzed. The soy components in our rodent diet produce a measurable background of daidzein and genistein in blood of female DA/Han rats, total isoflavones (aglycone plus conjugates) ranging between 90 and 290 ng/ml plasma. The ovariectomized animals kept on this chow, showed no signs of estrogenization of the reproductive tract (uterus, vagina), and responded normally to (xeno-)estrogen administration in a uterotrophic assay [J. Steroid Biochem. Mol. Biol., 73 (2000) 1]. Moreover, ovariectomized Wistar rats on our standard rodent diet (Ssniff R/M H) had lower uterine weights than animals kept on the isoflavone-free (solvent extracted) chow; both groups of rats responded to genistein administration with an increase in uterine weights. These results suggest that--albeit the sensitivity of the rodent uterotrophic assay is not reduced by the use of a diet containing soy isoflavones at commonly encountered levels--attention should be given to a variable dietary phytoestrogen background. PMID- 11869824 TI - Estradiol, testosterone, dehydroepiandrosterone and androstenedione: novel derivatives and enantiomers. Interactions with rat liver microsomal cytochrome P450 and antioxidant/radical scavenger activities in vitro. AB - Interactions of 27 steroids, among them 17 derivatives such as ethers, sulfates and amidosulfonates derived from 17 beta- and 17 alpha-estradiol, from testosterone and alpha- and beta-dihydrotesosterone and from dehydroepiandrosterone with rat liver microsomal cytochromes P450 (P450) were investigated in vitro by assessing binding to P450 and effects on P450 mediated monooxygenase functions as measured by different model reactions: ethoxyresorufin O-deethylation (EROD), ethoxycoumarin O-deethylation (ECOD) and ethylmorphine N demethylation (EMND). With the exception of 17 alpha-estradiol-3 dimethylamidosulfonate, estrone, its -3-methylether and -3-amidosulfonate and testosterone, all other steroids displayed type I or reverse type I binding to P450. All steroids inhibited EROD activity in micromolar concentrations. An additional strong inhibition of ECOD and EMND activities was only demonstrated for the androgens and progestins. Estriol, estrone and mestranol displayed less inhibitory actions on the model reactions than estradiol. No major differences in comparison to the parent compounds were noted with the other derivatives. The only exceptions were 17 beta-(8,9-dehydro-14 alpha,15 alpha-methylene)estradiol, which displayed stronger effects than estradiol, and dehydroepiandrosterone-3 sulfate, which was less effective than dehydroepiandrosterone. Possible antioxidant properties of the steroids were examined by the stimulated lipid peroxidation (LPO), H2O2 production, and lucigenin (LC) and luminol (LM) amplified chemiluminescence (CL) using rat liver microsomes. Additionally, the influence on rat whole blood chemiluminescence (WB-CL) was assessed. All the estrogens, but not their methylethers and amidosulfonates inhibited LPO in micromolar concentrations. The effects on the other oxidase model reactions or on WB-CL were less distinct. Only ethinylestradiol and 17 beta-(8,9-dehydro-14 alpha,15 alpha-methylene)estradiol displayed a strong inhibitory action on all model reactions. With the exception of dehydroepiandrosterone-3-sulfate, which in general had only weak effects, the androgen and progestin derivatives, in contrast, strongly decreased H2O2 formation and LM- and LC-CL, but were mostly ineffective on LPO and WB-CL. PMID- 11869826 TI - Myopathy and rhabdomyolysis with lipid-lowering drugs. AB - Drug-induced myopathy and rhabdomyolysis are rare adverse drug reactions (ADR). They have been seen after the introduction of modern lipid-lowering drugs more regularly. The first description after medication with clofibrate dates back to 1968. Apparently, all fibrates can induce myopathy. It usually starts after a few days of medication, or after prolonged use, showing muscle weakness and/or pain. Concomitantly, the enzyme creatininephosphokinase (CPK) is raised dramatically. Muscular necrosis can follow leading secondarily to kidney failure, and eventually to death. For the class of statins, myopathy was more often seen after their introduction, and it became their most feared adverse effect, especially in combination of statins with other drugs (mibefradil, gemfibrozil, cyclosporin). In animal models the evolution of the disease and the mechanism of action may be elucidated. Though strong epidemiological data are lacking, the incidence of myopathy is probably similar for all lipid-lowering drugs and is in the range of 0.1-0.5% with monotherapy, increasing to 0.5-2.5% with combination therapy. Severe cases of rhabdomyolysis are rarer, but may have a significant mortality. The market success of cerivastatin within a short period has led to 100s of myopathies and some dozens of deaths. Though interactions on metabolism and ensuing high plasma levels can partially explain myopathy as intoxication, there are strong indications that other (endocrine, metabolic, genetic) factors might play a role in the pathophysiology. The patient population at risk should better be defined and withheld from myopathy-inducing drugs. PMID- 11869827 TI - The influence of deferiprone (L1) and deferoxamine on iron and essential element tissue level and parameters of oxidative status in dietary iron-loaded mice. AB - The seven week feeding of a diet enriched with 0.5% TMH-ferrocene to male mice was used in this study to produce an iron-overload model in experimental animals for evaluating the effect of deferoxamine (DFO) and deferiprone (L1) on tissue stored iron, induced lipid peroxidation (LP) and parameters of oxidative status. The iron concentration in the liver reached 600% of the level in control animals. The administration of seven doses of deferoxamine (DFO) i.p. and deferiprone (L1) p.o. (0.72 mmol/kg b.w., every 48 h) during 9th and 10th week significantly decreased the liver, kidneys and heart iron level in both iron-loaded and control mice. The DFO and L1 treatment also equally attenuated lipid peroxidation and increased the GSH level in the liver of iron loaded mice. The glutathione peroxidase (GSH-Px) activity and catalase activity were not affected by iron loading, however, both DFO and L1 caused a decrease of GSH-Px activity. PMID- 11869828 TI - Relative potency of synthetic analogs of Ptychodiscus brevis toxin in depressing synaptic transmission evoked in neonatal rat spinal cord in vitro. AB - Effects of Ptychodiscus brevis toxin (PbTx) analogs on the spinal synaptic transmission in neonatal rats in vitro were evaluated. PbTx1/PbTx2 had aromatic groups and PbTx3/PbTx4 had aliphatic groups. All the analogs depressed monosynaptic reflex (MSR) and polysynaptic reflex (PSR) in a concentration dependent manner. The maximal depression of MSR (75% from initial) and PSR (96%) was at 84 microM for PbTx1. Concentration to produce 25% inhibition from initial (IC25) by PbTx1 for MSR and PSR was < or =2.8 microM. The maximal depression of MSR (80%) was at 96 microM and PSR (100%) was at 32 microM by PbTx2. IC25 for MSR and PSR were 5.5 microM and <3.2 microM, respectively. PbTx3 decreased MSR by 25% maximally (=IC25) at 36 microM. The depression of PSR fluctuated and was maximal (75%) at 108 microM and IC25 was 6.2 microM. PbTx4 depressed MSR and PSR at the maximum of 35% at 32 microM and IC25 for MSR was 8.3 microM and for PSR was 35 microM. Rank order of potency of toxins for depressing MSR was PbTx1>PbTx2>>PbTx4>PbTx3; and for PSR it was PbTx2>PbTx1>PbTx3>>PbTx4. Results indicate that the toxins having aromatic groups exhibited greater neurotoxicity. PMID- 11869829 TI - In vitro and in vivo attenuation of experimental cyanide poisoning by alpha ketoglutarate. AB - Treatment of cyanide poisoning generally includes methemoglobin forming agents, like amyl nitrite and/or sodium nitrite (SN), in combination with sodium thiosulphate (STS). However, in many instances of cyanide poisoning, use of nitrites are contraindicated due to their strong vasoactive properties. alpha Ketoglutarate (alpha-KG) antagonises cyanide by cyanohydrin formation and is considered a promising antidote for cyanide poisoning. In the present study, pre treatment (30 min) and simultaneous treatment (0 min) of alpha-KG (5 mM) was found to confer significant protection against 5 mM potassium cyanide (KCN) induced cytotoxicity in rat thymocytes as measured by eosin Y exclusion and leakage of intracellular lactate dehydrogenase (LDH), but could not prevent the mitochondrial dysfunction (MTT assay), depletion of cellular GSH (reduced glutathione) and DNA damage. The post-treatment (5 or 30 min) of alpha-KG did not offer any protection on any of the above parameters. Results of in vitro studies were also supported by in vivo data. Pre-treatment of peroral (p.o.) alpha-KG (0.125-2.0 g/kg) exhibited dose and time dependent effects and was found to be effective even when given upto 60 min prior to KCN (p.o.). Addition of STS significantly enhanced the protective efficacy of alpha-KG at all the doses and time intervals. A 10 min pre-treatment of alpha-KG increased the LD(50) of KCN 7.6-fold, which was further increased to 25.6-fold by the addition of both SN and STS. Simultaneous treatment of alpha-KG (2.0 g/kg) increased the LD50 of KCN 5.4 fold which was increased to 18.1-fold by the addition of STS. However, addition of SN did not confer any additional protection. In the presence of SN+STS, a decrease in the dose of alpha-KG exhibited a dose-dependent decrease in protection, but still a >10-fold protection could be observed at 1.0 g/kg dose of alpha-KG. Considering the efficacy and safety of peroral alpha-KG, a promising treatment regimen consisting of alpha-KG+STS or alpha-KG+SN+STS is proposed, depending upon the individual situation. PMID- 11869831 TI - The effect of lofepramine and other related agents on the motility of Tetrahymena pyriformis. AB - Tricyclic antidepressants (TCAs) were introduced almost 50 years ago. Whilst there is no doubt that TCAs are effective in treating depression, they are also more cardiotoxic when taken in overdose than other antidepressant groups. Lofepramine is a more recently introduced modified TCA, which in animals and man has low toxicity when compared to older TCAs. Paradoxically, lofepramine is extensively metabolised to desipramine, which has considerable toxicity, both experimentally and in overdose. The toxicity of such compounds is attributed, in part, to a membrane stabilising effect (MSA) on cell membranes. This MSA causes gross effects to the cell structure and in turn, normal cell activity. The aim of this study was to compare the MSA of lofepramine with that of desipramine and amitriptyline in order to see if this might help to explain the low toxicity of lofepramine. The local anaesthetic agent lignocaine was also studied for comparison. Each compound was enclosed in a beta-cyclodextrin to increase its solubility in aqueous medium. The extent of MSA was determined as a measure of the effect on the swimming speed of the protozoan Tetrahymena pyriformis using a video image analysis system. The IC50s for the various drugs were then correlated with their respective octanol-water partition coefficient values (Pow). Amitriptyline had an IC50 of 1.26+/-0.29 mM, desipramine 75.99+/-14.40 mM, while lofepramine had an IC50 of 357.40+/-25.00 mM. Lignocaine had an IC50 of 85.73+/ 18.30 mM. There was also a significant correlation between the IC50 values and the Pow values. PMID- 11869832 TI - Enzymes involved in the detoxification of organophosphorus, carbamate and pyrethroid insecticides through hydrolysis. AB - The most employed insecticides for indoor and agriculture purposes belong to carbamates, pyrethroid or organophosphates. The chemical structures of these three groups correspond to carbamic, carboxylic and triphosphoric esters. Technical monographs suggest that the hydrolysis of ester bonds of carbamates and pyrethroids plays an important role in the detoxification of these compounds. However, detailed studies about enzymes hydrolysing carbamates and pyrethroids in vertebrates are not available. Certain carbamate hydrolysing activities are associated to serum albumin. Phosphotriesterases, being of an unknown physiological role, hydrolyse (in some cases stereospecifically) organophosphorus insecticides (OP). Phosphotriesterases have been found in a multitude of species, from mammals to bacteria. A phosphotriesterase activity, EDTA-resistant, has been detected in serum albumin. Phosphotriesterases in serum of mammals display polymorphisms. Phosphotriesterases offer applications in therapy of organophosphorus poisonings, in biodegradation and bioremedation of organophosphates. Similar studies should be developed with enzymes hydrolysing pyrethroids and carbamate insecticides. Such studies will improve the knowledge of the detoxification routes in non-target species and will help to design specific and safer carbamate and pyrethroid insecticides. PMID- 11869830 TI - Interactions of the neurotoxin 6-hydroxydopamine with glyceraldehyde-3-phosphate dehydrogenase. AB - The neurotoxin 6-hydroxydopamine (6-OHDA) oxidised sulphydryl groups in glyceraldenyde-3-phosphate dehydrogenase (GAPDH) with loss of the dehydrogenase activity of the enzyme. 5-Hydroxydopamine behaved similarly at higher concentrations. This oxidation was accompanied by a transient rise in the acyl phosphatase (non-phosphorylating) activity of GAPDH. Treatment with arsenite resulted in loss of the acyl phosphatase activity and restoration of the dehydrogenase activity, consistent with the process involving sulphydryl-group oxidation to the corresponding sulphenate. Prolonged incubation with 6-OHDA resulted in a loss of both enzyme activities. Arsenite was unable to reverse this inhibition, indicating further oxidation, perhaps to sulphinate and sulphonate, to have occurred. RNA, but not DNA, was a partial inhibitor of both the dehydrogenase and acyl phosphatase activities of GAPDH, whereas DNA (both single- and double-stranded) was ineffective. Both single-stranded DNA and RNA inhibited the esterase activity of GAPDH, an activity that requires the absence of NAD+, in a process that was relieved at higher polynucleotide concentrations. Except at very high concentrations (1 mM), treatment of GAPDH 6-OHDA was ineffective at abolishing its to single-stranded DNA. Since GAPDH is an essential enzyme in glycolysis and also plays a role in apoptotic cell death, these results suggest that the effects on this enzyme may contribute to the neurotoxicity of 6-OHDA. PMID- 11869833 TI - The enhanced bladder cancer susceptibility of NAT2 slow acetylators towards aromatic amines: a review considering ethnic differences. AB - Human bladder cancer may be caused by exposure to aromatic amines. The polymorphic enzyme N-acetyltransferase 2 (NAT2) is involved in the metabolism of these compounds. Two classical studies on chemical workers in Europe, exposed in the past to aromatic amines like benzidine, unambiguously showed that the slow acetylator status is a genetic risk factor for arylamine-induced bladder cancer. In the former benzidine industry in Huddington, Great Britain, 22 of 23 exposed cases with bladder cancer, but only 57% of 95 local controls without bladder cancer were of the slow acetylator phenotype. In Leverkusen, Germany, 82% of 92 benzidine-exposed chemical workers with bladder cancer were of the slow acetylator phenotype, whereas only 48% of 331 chemical workers who had worked at that plant were of the slow acetylator phenotype. This is in line with several smaller studies, which also show an over-representation of the slow acetylator status in formerly arylamine-exposed subjects with bladder cancer. Some of these studies included also subjects that were exposed to aromatic amines by having applied dyes, paints and varnishes. These European findings are in contrast to a large study on Chinese workers occupationally exposed to aromatic amines. In this study, only five of 38 bladder cancer cases occupationally exposed to arylamines were of the slow acetylator genotype. This is much lower than the ratio of slow acetylators to the general population in China. This points to different mechanisms of susceptibility for bladder cancer upon exposure to aromatic amines between European (Caucasian) and Chinese populations. PMID- 11869834 TI - Renal toxicity after chronic inhalation exposure of rats to trichloroethylene. AB - Male Long-Evans rats were exposed to 0 (controls) or 500 ppm trichloroethylene (TRI) for 6 months, 6 h daily, and 5 days a week. The TRI metabolites trichloroethanol (TCE) in blood and trichloroacetic acid (TCA) in urine were measured. Specific parameters related to the renal damage were determined in urine [biomarker for glomerular damage: high molecular weight proteins (HMW), albumin (ALB); for proximal tubular damage: N-acetyl-beta-D-glucosaminidase (NAG), low-molecular-weight-proteins (LMW)]. Significantly increased concentrations of NAG and LMW in urine of exposed rats were detected. No DNA strand breaks in kidney cells could be detected using the comet assay, and histological examinations were performed. Histological alterations were observed in glomeruli and tubuli of exposed rats. The release of biomarkers for nephrotoxicity suggested alterations preferably in the proximal tubules of the exposed rats. PMID- 11869835 TI - Possible impact of human CYP2E1 polymorphisms on the metabolism of acrylonitrile. AB - Case reports of human accidental poisonings point to significant individual differences in human acrylonitrile metabolism and toxicity. A cohort of 59 persons with industrial handling of low levels of acrylonitrile has repetitively been studied from 1994 through 1999 as part of a medical surveillance programme. The analyses included adduct determinations of N-terminal N-(cyanoethyl)valine in haemoglobin and genotypings of the following cytochrome P-450 2E1 (CYP2E1) polymorphisms: G-1259C and C-1019T (two subjects heterozygous), A-316G (three subjects heterozygous), T-297A (15 subjects heterozygous), G-35T (eight subjects heterozygous), G4804A (two subjects heterozygous), T7668A (six subjects heterozygous). N-(Cyanoethyl)valine adduct levels were, if any, only slightly influenced by smoking and mainly determined by the external acrylonitrile exposures. The individual means and medians of N-(cyanoethyl)valine levels over the entire observation period were compared with the CYP2E1 variants (Wilcoxon rank sum test). No influences of the investigated CYP2E1 polymorphisms on the N (cyanoethyl)valine levels appeared at the 5% level. However, there was a trend, at a level of P approximately 0.1, pointing to higher acrylonitrile-specific adduct levels in persons with the A-316G mutation. Higher adduct levels would be compatible with a slower CYP2E1-mediated metabolism of acrylonitrile and with lower extents of toxification to cyanide. PMID- 11869836 TI - Na(+)/H(+) exchange inhibitors for cardioprotective therapy: progress, problems and prospects. AB - Extensive pre-clinical work indicates that inhibition of the sarcolemmal Na(+)/H(+) exchanger (NHE) affords significant protection to myocardium subjected to ischemia and reperfusion, predominantly through reduced intracellular accumulation of Na(+) and consequently Ca(2+). In contrast, recent clinical studies with the NHE inhibitors cariporide and eniporide in patients with evolving myocardial infarction (MI) and those at risk of MI have provided mixed and somewhat contradictory data. The experimental evidence suggests that the key mechanism through which NHE inhibitors afford protection consists in slowing the progression of myocardial injury during ischemia and thereby enhancing myocardial salvage by reperfusion. It follows from this that, to obtain maximum cardioprotective benefit, 1) the NHE inhibitor must be present in jeopardized myocardium, at a concentration sufficient to inhibit NHE activity, before (or as soon as possible after) the onset of ischemia, and 2) ischemia must be terminated by timely reperfusion. Thus, in the GUARDIAN trial, the cardioprotective efficacy of cariporide was limited to the subset of high-risk patients who underwent coronary artery bypass graft surgery, in whom both prerequisites could be readily fulfilled. In contrast, no cardioprotective benefit was observed in the ESCAMI trial, in which eniporide was administered late as an adjunct to reperfusion therapy in patients with evolving MI. Ongoing clinical studies will determine whether NHE inhibition will find therapeutic application in the setting of cardiac surgery, while pre-clinical investigations continue to test the potential of NHE inhibitors in the treatment of other cardiovascular diseases such as heart failure. PMID- 11869837 TI - Repeated sauna treatment improves vascular endothelial and cardiac function in patients with chronic heart failure. AB - OBJECTIVES: The purpose of this study was to determine the mechanism by which 60 degrees C sauna treatment improves cardiac function in patients with chronic heart failure (CHF). BACKGROUND: We have previously reported that repeated 60 degrees C sauna treatment improves hemodynamic data and clinical symptoms in patients with CHF. We hypothesized that the sauna restores endothelial function and then improves cardiac function. METHODS: Twenty patients (62 plus minus 15 years) in New York Heart Association (NYHA) functional class II or III CHF were treated in a dry sauna at 60 degrees C for 15 min and then kept on bed rest with a blanket for 30 min, daily for two weeks. Ten patients with CHF, matched for age, gender and NYHA functional class, were placed on a bed in a temperature controlled (24 degrees C) room for 45 min as the nontreated group. Using high resolution ultrasound, we measured the diameter of the brachial artery at rest and during reactive hyperemia (percent flow-mediated dilation, %FMD: endothelium dependent dilation), as well as after sublingual administration of nitroglycerin (%NTG: endothelium-independent dilation). Cardiac function was evaluated by measuring the concentrations of plasma brain natriuretic peptide (BNP). RESULTS: Clinical symptoms were improved in 17 of 20 patients after two weeks of sauna therapy. The %FMD after two-week sauna treatment significantly increased from the baseline value, whereas the %NTG-induced dilation did not. Concentrations of BNP after the two-week sauna treatment decreased significantly. In addition, there was a significant correlation between the change in %FMD and the percent improvement in BNP concentrations in the sauna-treated group. In contrast, none of the variables changed at the two-week interval in the nontreated group. CONCLUSIONS: Repeated sauna treatment improves vascular endothelial function, resulting in an improvement in cardiac function and clinical symptoms. PMID- 11869838 TI - Heart failure in a cold climate. Seasonal variation in heart failure-related morbidity and mortality. AB - OBJECTIVES: This study was done to determine whether seasonal variation exists in hospitalizations and deaths due to heart failure (HF) and to examine possible contributors to such variability. BACKGROUND: Although seasonal variation in the incidence of acute myocardial infarction and sudden death is well recognized, it is less well documented in HF. METHODS: We used the linked Scottish Morbidity Record scheme, which provides individualized morbidity and mortality data for the entire Scottish population. RESULTS: Between 1990 and 1996, there were a total of 75,452 male and 81,269 female hospitalizations related to HF in Scotland, with an average rate of admissions per 100,000 population of 8.4 and 8.5 per day, respectively. Significantly more admissions occurred in winter compared to summer (p < 0.0001). In women, the peak rate of admission occurred in December (12% more than average) and the lowest rate in July (7% less than average) (odds ratio [OR] 1.14, p < 0.001). The respective figures for men were 6% more, 8% less (OR 1.16, p < 0.001). In both genders, the greatest variation occurred in those aged >75 years---peak winter rates being 15% to 18% higher than average. There was also a winter peak in concomitantly coded respiratory disease; this seasonal excess accounted for approximately one-fifth of the winter increment in HF hospitalizations. Seasonal variation in mortality was also seen in these patients. The number of male deaths in December was 16% higher, and in July 7% lower, than average (OR 1.25, p < 0.001). In women, the equivalent figures were 21% higher (January) and 14% lower (July) (OR 1.21, p < 0.001). Again, the greatest variation occurred in those aged >75 years---peak rates being 23% to 35% higher than average. CONCLUSIONS: There is substantial seasonal variation in HF hospitalizations and deaths, particularly in the elderly. Approximately one-fifth of the winter excess in admissions is attributable to respiratory disease. Extra vigilance in patients with HF is advisable in winter, as is immunization against pneumococcus and influenza. PMID- 11869839 TI - Gradual reactivation over time of vascular tissue angiotensin I to angiotensin II conversion during chronic lisinopril therapy in chronic heart failure. AB - OBJECTIVES: This study was designed to fully characterize vascular tissue angiotensin I (AI)/angiotensin II (AII) conversion changes over time in vivo in humans during chronic angiotensin-converting enzyme (ACE) inhibitor therapy. BACKGROUND: Plasma AII does not remain fully suppressed during chronic ACE inhibitor therapy. However, the plasma renin angiotensin system (RAS) might be dissociated from the vascular tissue RAS. We therefore set out to characterize the time course of vascular RAS reactivation during chronic ACE inhibitor therapy. METHODS: Vascular AI/AII conversion was studied in patients with chronic heart failure (CHF) taking chronic lisinopril therapy by the differential infusion of AI and AII into the brachial artery. A cross-sectional study was done to see whether there were differences in vascular AI/AII conversion according to New York Heart Association (NYHA) class. A second longitudinal study followed 28 patients with NYHA I to II CHF serially over 18 months to see whether vascular ACE inhibition was progressively lost with time despite ACE inhibitor therapy. A third study examined whether increasing the dose of lisinopril affected subsequent vascular ACE inhibition. RESULTS: In the cross-sectional study, vascular AI-to-AII conversion was significantly reduced in NYHA class III compared with class I/II (p < 0.05). In the longitudinal study, vascular ACE inhibition was significantly reduced at 18 months as compared with baseline (p < 0.001), suggesting gradual reactivation of vascular ACE in CHF over time. In the third study, tissue ACE inhibition could be restored by increasing the ACE inhibitor dose. CONCLUSIONS: Vascular AI/AII conversion reactivates over time during chronic ACE inhibitor therapy even if the CHF disease process is clinically stable. It also occurs as the CHF disease process progresses. Even if vascular AI/AII conversion has reactivated, it can be suppressed by increasing the dose of the ACE inhibitor. PMID- 11869840 TI - Activation of vascular tissue angiotensin-converting enzyme (ACE) in heart failure. Effects of ACE inhibitors. PMID- 11869841 TI - Implantable defibrillator event rates in patients with idiopathic dilated cardiomyopathy, nonsustained ventricular tachycardia on Holter and a left ventricular ejection fraction below 30%. AB - OBJECTIVES: This study investigated the incidence of appropriate implantable cardioverter defibrillator (ICD) interventions for ventricular tachycardia (VT) or ventricular fibrillation (VF) in patients with idiopathic dilated cardiomyopathy (IDC) and nonsustained VT in the presence of a left ventricular ejection fraction below 30%, versus in patients with syncope and patients with a history of VT or VF. BACKGROUND: To date, only limited information is available about the prophylactic use of ICDs in patients with IDC. METHODS: From January 1993 to July 2000, 101 patients with IDC underwent implantation of ICDs with electrogram storage capability at our institution. Patients were placed into one of three groups according to their clinical presentation: asymptomatic or mildly symptomatic nonsustained VT in the presence of a left ventricular ejection fraction < or = 30% (49 patients, prophylactic group), unexplained syncope or near syncope (26 patients, syncope group) and a history of sustained VT or VF (26 patients, VT/VF group). RESULTS: During 36 +/- 22 months follow-up, 18 of 49 patients (37%) in the prophylactic group received appropriate shocks for VT or VF, compared with 8 of 26 patients (31%) in the syncope group and with 9 of 26 patients (35%) of the VT/VF group. Multivariate Cox analysis of baseline clinical variables identified left ventricular ejection fraction, atrial fibrillation and a history of sustained VT or VF as predictors for appropriate ICD interventions during follow-up. CONCLUSIONS: Patients with IDC and prophylactic ICD implantation for nonsustained VT in the presence of a left ventricular ejection fraction < or = 30% had an incidence of appropriate ICD interventions similar to that of patients with a history of syncope or sustained VT or VF. These findings indicate that ICDs may have a role in not only secondary but also primary prevention of sudden death in IDC. PMID- 11869842 TI - Primary prevention of sudden death using ICD therapy: incremental steps. PMID- 11869843 TI - Initial experience with an implantable cardioverter-defibrillator incorporating cardiac resynchronization therapy. AB - OBJECTIVES: The purpose of this study was to evaluate an implantable cardioverter defibrillator (ICD) incorporating biventricular pacing. BACKGROUND: Biventricular pacing improves the symptoms of heart failure, a frequent problem in ICD recipients. METHODS: This prospective multicenter study evaluated the safety and efficacy of an ICD with biventricular pacing. RESULTS: A total of 84 patients with a standard ICD indication, symptomatic heart failure, left ventricular (LV) ejection fraction <35% and a QRS duration >130 ms were included in the trial. In 81 of 84 patients the LV lead was successfully implanted. Patients significantly improved in the 6-min hall-walk test (baseline 304 plus minus 131 m, three months 397 plus minus 142 m, p < 0.001), quality of life (baseline 38.9 plus minus 21.2, three months 26.5 plus minus 21.2, p < 0.001) and the New York Heart Association (NYHA) classification (baseline 2.8 plus minus 0.6, three months 2.2 plus minus 0.5). Left ventricular end-diastolic (from 79.6 plus minus 13.0 mm to 73.6 plus minus 12.9 mm, p = 0.002) and end-systolic (from 68.3 plus minus 13.5 mm to 63.9 plus minus 12.9 mm, p < 0.001) diameter decreased, and fractional shortening increased (from 16 plus minus 6% to 18 plus minus 6%, p = 0.018). Of the patients 26 experienced 472 episodes of spontaneous sustained ventricular tachyarrhythmias. All episodes were successfully terminated except for 16 episodes occurring in a patient with incessant ventricular tachycardia. Biventricular antitachycardia pacing was more effective than right ventricular antitachycardia pacing (p < 0.001). During follow-up (median 185 days, range 12 to 344 days) five patients died from progressive heart failure. CONCLUSIONS: Incorporating biventricular pacing in an ICD is feasible and leads to an improvement of heart failure symptoms. Therefore, this therapy may become an option for patients who need ICD therapy in the presence of severe heart failure. PMID- 11869844 TI - Effect of nesiritide versus dobutamine on short-term outcomes in the treatment of patients with acutely decompensated heart failure. AB - OBJECTIVES: This study was designed to determine whether nesiritide, administered for acute decompensated congestive heart failure (CHF), affects healthcare costs by hospital length of stay (LOS), readmissions and short-term mortality, compared to dobutamine. BACKGROUND: Dobutamine is a commonly used inotropic treatment for CHF. Although dobutamine may have favorable hemodynamic and symptomatic effects, its use may be associated with side effects such as tachycardia, cardiac arrhythmias and myocardial ischemia. Nesiritide (B-type natriuretic peptide) is a new intravenous (IV) drug that produces hemodynamic and symptomatic improvement through balanced vasodilatory effects, neurohormonal suppression and enhanced natriuresis and diuresis. METHODS: From an open-label randomized study of nesiritide versus standard care (SC) in patients with CHF requiring hospitalization, we compared short-term outcome data from patients given nesiritide (0.015 or 0.03 microg/kg per min) with a subgroup of SC patients given dobutamine. A total of 261 patients are included in this analysis. RESULTS: Compared to dobutamine, both nesiritide doses were administered for a shorter total duration (p < 0.001), and the total duration of all IV vasoactive therapy (including study drug) was also shorter (p less-than-or-equal 0.012). Although there was no difference in LOS, there was a trend toward decreased readmissions in the two nesiritide groups (8% and 11%, respectively, vs. 20% in the dobutamine group). Six-month mortality was lower in the nesiritide groups. CONCLUSIONS: Treatment of decompensated CHF with nesiritide may lead to lower healthcare costs and reduced mortality compared to treatment with dobutamine. PMID- 11869845 TI - The role of vitronectin receptor (alphavbeta3) and tissue factor in the pathogenesis of transplant coronary vasculopathy. AB - OBJECTIVES: This study was undertaken to test the hypothesis that transplant coronary vasculopathy (CV) is associated with increased myocardial protein expression of both tissue factor (TF) and integrin alphavbeta3. BACKGROUND: The vitronectin receptor (integrin alphavbeta3) and TF have recently been found to play a key role in apoptotic cell death and vascular endothelial cell injury. METHODS: A total of 77 heart transplant recipients underwent simultaneous endomyocardial biopsy and intravascular ultrasound (IVUS) at one year of transplant. Patients with pre-existing donor coronary atherosclerosis (n = 35) or with acute rejection (grade >1A, n = 10) at the time of the IVUS were excluded from the analysis. The remaining 32 patients constitute the cohort of the present study. A computerized biopsy score was derived based on the duration and severity of cellular rejection. Both TF and alphavbeta3 expression in the heart biopsy specimens were evaluated by immunoperoxidase histochemistry and Western blot analysis. RESULTS: Patients with CV (n = 24) had increased expression of alphavbeta3 (2.7-fold, p = 0.003) and TF (7.9-fold, p = 0.04) compared with patients without evidence of vasculopathy (n = 8). In the absence of myocardial fibrosis, alphavbeta3 expression correlated significantly with the cellular rejection score (r = 0.58, p = 0.02). CONCLUSIONS: Transplant vasculopathy is associated with increased expression of both TF and alphavbeta3. The significant correlation of alphavbeta3 with cellular rejection suggests an important role for this integrin in serving as a mechanistic link between cellular rejection and vasculopathy. PMID- 11869846 TI - Rebound thrombin generation after heparin therapy in unstable angina. A randomized comparison between unfractionated and low-molecular-weight heparin. AB - OBJECTIVES: This study compared rebound coagulation in patients with acute coronary syndrome patients after discontinuation of unfractionated heparin (UFH) or low-molecular-weight heparin (LMWH). BACKGROUND: Up to a quarter of patients hospitalized for unstable angina experience recurrent ischemia after discontinuation of UFH or LMWH therapy, which may be the result of rebound coagulation activation and subsequent thrombosis. It is unknown whether UFH and LMWH differ in this respect. METHODS: We randomized 71 patients admitted with unstable angina to intravenous UFH or subcutaneous LMWH (dalteparin) and measured plasma markers of coagulation before, during, and after treatment. RESULTS: A complete series of measurements was obtained in 59 patients. Plasma prothrombin fragment 1+2 (F(1+2)) levels decreased in both groups during treatment. After loss of therapeutic plasma drug levels, F(1+2) increased (within 3 h) to a maximum level at 12 to 24 h that was higher than before or during treatment in both groups (p < 0.0001). In both groups, F(1+2) levels remained higher than pretreatment up to 24 h after discontinuation. Similarly, thrombin-antithrombin (TAT) levels exceeded treatment and pretreatment levels, at a slower rate after dalteparin than after UFH. However, after dalteparin a higher peak value of TAT was observed. CONCLUSIONS: Rebound coagulation activation occurs within hours after discontinuation of both UFH and dalteparin. With both drugs, thrombin generation is significantly greater after treatment than before or during treatment. A longer duration or weaning of treatment, or continuation with another anticoagulant treatment, may reduce rebound coagulation activation and ischemic events. PMID- 11869847 TI - Nitric oxide production by neutrophils obtained from patients during acute coronary syndromes: expression of the nitric oxide synthase isoforms. AB - OBJECTIVES: To analyze the differences in the nitric oxide (NO) forming system between neutrophils obtained from patients during unstable angina (UA) and during acute myocardial infarction (AMI). BACKGROUND: Neutrophils are involved in the regulation of thrombus formation through the release of active substances such as NO. Acute myocardial infarction is the result of an occlusive thrombus; unstable angina is attributed to intermittent thrombus formation. METHODS: We studied 49 patients admitted to hospital within 24 h after the onset of chest pain: 31 experienced AMI and 18 experienced UA. Acute myocardial infarction was defined as CK greater than two-fold the upper limit of normal value of biochemical laboratory, with CK-MB >10% total CK. Unstable angina was defined as transient ST segment changes without significant increases in CK and CK-MB. RESULTS: The amount of NO generated by neutrophils from AMI patients was significantly higher than that generated by neutrophils from UA patients. Neutrophils from UA and AMI patients showed low levels of endothelial-like NO synthase protein expression and a marked expression of the inducible NO synthase (iNOS) isoform. Although neutrophils from patients during acute coronary syndromes generated high amounts of NO, they did not demonstrate an increased ability to stimulate cyclic guanosine monophosphate (cGMP) synthesis in platelets. This lack of activity to release NO by neutrophils from patients during AMI was unrelated to a defect in the platelet cGMP-forming system; sodium nitroprusside, an exogenous NO donor, similarly increased cGMP levels in platelets from AMI patients and healthy donors. CONCLUSIONS: Neutrophils from patients during AMI and UA showed an increased production of NO and a marked expression of the iNOS isoform. However, NO released from these neutrophils showed a deficient functionality. These findings could have clinical implications because they show differences in thrombus growth in patients with UA versus patients with AMI. PMID- 11869848 TI - Primary angioplasty and selection bias inpatients presenting late (>12 h) after onset of chest pain and ST elevation myocardial infarction. AB - OBJECTIVES: This study was designed to compare the in-hospital outcome of patients presenting with >12 h from onset of chest pain and acute ST elevation myocardial infarction (AMI) who received either immediate invasive or conservative therapy. BACKGROUND: The benefits of fibrinolytic therapy diminish in patients presenting with AMI and onset of chest pain >12 h. Primary angioplasty has been suggested as a possible treatment for such patients, but they have been excluded from most trials of primary angioplasty. It remains unclear if an invasive treatment strategy is beneficial to these patients. METHODS: Patients presenting with >12 h of chest pain and AMI were identified from the National Registry of Myocardial Infarction 2 database. Patients receiving invasive therapy <6 h after hospital admission were compared with those receiving conservative therapy. Short-term outcomes were compared on the basis of the initial therapy received. To help control for baseline differences in the groups, patients were matched with controls by propensity score methodology. RESULTS: On preliminary analysis, in-hospital outcome was improved in terms of recurrent ischemia, angina, myocardial infarction and mortality in patients receiving initial invasive therapy (odds ratio [OR] = 0.67; 95% confidence interval [CI] 0.49 to 0.92 for mortality). After matching by propensity score, the mortality benefit persisted on bivariate analysis (3.5% vs. 5.0%, p = 0.036), though on multivariate analysis, only a non-significant but strong trend toward decreased mortality remained (OR = 0.73; 95% CI 0.53 to 1.01). CONCLUSIONS: Patients receiving early invasive therapy had lower risk features on presentation. Selection bias may play an important role in choosing these patients' course of treatment and their subsequent outcomes. Certain patients presenting with AMI and duration of chest pain >12 h may benefit from early invasive therapy. These patients could be characterized in a randomized trial. PMID- 11869849 TI - The impact of body mass index on short- and long-term outcomes inpatients undergoing coronary revascularization. Insights from the bypass angioplasty revascularization investigation (BARI). AB - OBJECTIVES: We sought to investigate the impact of body mass index (BMI) on short and long-term outcomes after initial revascularization with percutaneous transluminal coronary angioplasty (PTCA) or coronary artery bypass graft surgery (CABG). BACKGROUND: Equivocal results exist on the impact of BMI on the risk of in-hospital complications after PTCA or CABG, and no long-term mortality data exist from a large series of revascularized patients. METHODS: From the randomized series and observational registry of the Bypass Angioplasty Revascularization Investigation (BARI), 2,108 patients who had PTCA and 1,526 patients who had CABG were evaluated by taking their BMI at study entry. They were classified as follows: low (< 20 kg/m(2)), normal (20 to 24.9 kg/m(2)), overweight (25 to 29.9 kg/m(2)), class I obese (30 to 34.9 kg/m(2)) and class II/III obese (greater-than-or-equal 35 kg/m(2)). In-hospital complications and short- and long-term mortalities were compared between levels of BMI within each mode of initial revascularization. RESULTS: Among patients who had PTCA, each unit increase in BMI was associated with a 5.5% lower adjusted risk of a major in hospital event (death, myocardial infarction, stroke, coma); among patients who had CABG, no difference in the in-hospital outcome was observed according to BMI. In contrast, BMI was not associated with five-year mortality in the PTCA group; among the CABG group, adjusted relative risks of five-year cardiac mortality according to levels of BMI were 0.0 (low), 1.0 (normal), 2.02 (overweight), 3.16 (class I obese) and 4.85 (class II/III obese) (linear p < 0.001). CONCLUSIONS: Body mass index appears to have a differential impact on short- and long-term outcomes after coronary revascularization. These results underscore the need for further research to identify factors responsible for the apparent short-term protective effect of a higher BMI in patients undergoing PTCA and to study the impact of weight reduction on the long-term survival of obese patients undergoing CABG. PMID- 11869850 TI - Multicenter evaluation of carotid artery stenting with a filter protection system. AB - OBJECTIVES: The aim of this study was to evaluate the feasibility and safety of carotid artery stenting (CAS) with a filter protection system. BACKGROUND: Neurologic events linked to the embolization of particulate matter to the cerebral circulation may complicate CAS. Strategies designed to capture embolic particles during carotid intervention are being evaluated for their efficacy in reducing the risk of these events. METHODS: Between September 1999 and July 2001, a total of 162 patients (164 hemispheres) underwent CAS with filter protection (NeuroShield, MedNova Ltd., Galway, Ireland) according to prospective protocols evaluating the filter system at three institutions. RESULTS: Angiographic success was achieved in 162 of the procedures (99%) and filter placement was successful in 154 (94%) procedures. Carotid access was unsuccessful in two cases (1%) and filter placement in eight cases (5%). Of the latter, five procedures were completed with no protection and three were completed using alternative protection devices. On an intention-to-treat basis, the overall combined 30-day rate of all-stroke and death was 2% (four events: two minor strokes and two deaths). This includes one minor stroke in a patient with failed filter placement and CAS completed without protection. There was one cardiac arrhythmic death and one death from hyperperfusion-related intracerebral hemorrhage. There were no major embolic strokes. CONCLUSIONS: Carotid artery stenting with filter protection is technically feasible and safe. Early clinical outcomes appear to be favorable and need to be confirmed in a larger comparative study. PMID- 11869851 TI - Coronary microvascular spasm causes myocardial ischemia in patients with vasospastic angina. AB - OBJECTIVES: We aimed to test the hypothesis that coronary microvascular spasm (MVS) alone causes myocardial ischemia in patients with angina attributable to epicardial coronary spasm, and to determine whether there is a difference in clinical characteristics between those with and without microvascular spasm. BACKGROUND: Patients with "vasospastic angina" have epicardial coronary artery spasm, but it is unknown whether coronary microvessel disease also contributes to the occurrence of angina in these patients. METHODS: We studied 55 consecutive patients with angina in whom epicardial coronary spasm was provoked by intracoronary acetylcholine (ACH). RESULTS: In 14 patients (25.5%, Group 1), submaximal dose of ACH induced myocardial ischemia (chest pain, ischemic electrocardiogram changes, lactate production) without large epicardial spasm, suggesting the occurrence of coronary microvascular spasm. By contrast, the remaining 41 patients (Group 2) had evidence of myocardial ischemia only when epicardial spasm was angiographically demonstrated. The Group 1 patients were predominantly women (p < 0.05) and had a history of prolonged (>30 min) chest pain (p < 0.05), whereas the Group 2 patients were more likely men and smokers (p < 0.01). CONCLUSIONS: Myocardial ischemia most probably due to coronary MVS was demonstrated in a sizable portion of patients with epicardial vasospasm, preferentially in women having both typical and prolonged anginal pain. The result suggests that coronary microvascular disease may also contribute to angina in patients with "vasospastic angina." PMID- 11869852 TI - Prognostic value of coronary blood flow velocity and myocardial perfusion in intermediate coronary narrowings and multivessel disease. AB - OBJECTIVES: This study aimed to investigate the roles of intracoronary derived coronary flow velocity reserve (CFVR) and myocardial perfusion scintigraphy (single photon emission computed tomography, or SPECT) for management of an intermediate lesion in patients with multivessel coronary artery disease. BACKGROUND: Evaluation of the functional significance of intermediate coronary narrowings (40% to 70% diameter stenosis) is important for clinical decision making and risk stratification. METHODS: In a prospective, multicenter study, SPECT was performed in 191 patients with stable angina and multivessel disease and scheduled for angioplasty (percutaneous transluminal coronary angioplasty, or PTCA) of a severe coronary narrowing. Coronary flow velocity reserve was determined selectively distal to an intermediate lesion in another artery using a Doppler guidewire. Percutaneous transluminal coronary angioplasty of the intermediate lesion was deferred when SPECT was negative or CFVR greater-than-or equal 2.0. Patients were followed for one year to document major cardiac events (death, infarction, revascularization), related to the intermediate lesion. RESULTS: Reversible perfusion defects were documented in the area of the intermediate lesion in 30 (16%) patients; CFVR was positive in 46 (24%) patients. Percutaneous transluminal coronary angioplasty of the intermediate lesion was deferred in 182 patients. During follow-up, 19 events occurred (3 myocardial infarctions, 16 revascularizations). Coronary flow velocity reserve was a more accurate predictor of cardiac events than was SPECT; relative risk: CFVR 3.9 (1.7 to 9.1), p < 0.05; SPECT 0.5 (0.1 to 3.2), p = NS. Multivariate analysis revealed CFVR as the only significant predictor for cardiac events. CONCLUSIONS: Deferral of PTCA of intermediate lesions in multivessel disease is safe when CFVR greater than-or-equal 2.0 (event rate 6%). This selective evaluation of coronary lesion severity during cardiac catheterization allows a more accurate risk stratification than does SPECT, which is important for clinical decision making in this patient cohort. PMID- 11869853 TI - Coronary flow studies for risk stratification in multivessel disease. A physiologic bridge too far? PMID- 11869854 TI - Assessment of coronary flow reserve: comparison between contrast-enhanced magnetic resonance imaging and positron emission tomography. AB - OBJECTIVES: The study compared flow reserve indices by magnetic resonance imaging (MRI) with quantitative measures of coronary angiography and positron emission tomography (PET). BACKGROUND: The noninvasive evaluation of myocardial flow by MRI has recently been introduced. However, a comparison to quantitative flow measurement as assessed by PET has not been reported in patients with coronary artery disease (CAD). METHODS: Two groups of healthy volunteers and 25 patients with angiographically documented CAD were examined by MRI and PET at rest and during adenosine stress. Dynamic MRI was performed using a multi-slice ultra-fast hybrid sequence and a rapid gadolinium-diethylenetriaminepenta-acetic acid bolus injection (0.05 mmol/l). Upslope and peak-intensity indices were regionally determined from first-pass signal intensity curves and compared to N-13 ammonia PET flow reserve measurements. RESULTS: In healthy volunteers, the upslope analysis showed a stress/rest index of 2.1 plus minus 0.6, which was higher than peak intensity (1.5 plus minus 0.3), but lower than flow reserve by PET (3.9 plus minus 1.1). Localization of coronary artery stenoses (> 75%, MRI < 1.2), based on the upslope index, yielded sensitivity, specificity and diagnostic accuracy of 69%, 89% and 79%, respectively. Upslope index correlated with PET flow reserve (r = 0.70). A reduced coronary flow reserve (PET < 2.0, MRI < 1.3) was detected by the upslope index with sensitivity, specificity and diagnostic accuracy of 86%, 84% and 85%, respectively. CONCLUSIONS: Magnetic resonance imaging first-pass perfusion measurements underestimate flow reserve values, but may represent a promising semi-quantitative technique for detection and severity assessment of regional CAD. PMID- 11869855 TI - Pharmacologic preconditioning of estrogen by activation of the myocardial adenosine triphosphate-sensitive potassium channel in patients undergoing coronary angioplasty. AB - OBJECTIVES: The purpose of this study was to determine whether administration of estrogen produces cardioprotective effects in patients undergoing coronary angioplasty. BACKGROUND: We have previously demonstrated that estrogen can provide cardioprotection by activating the mitochondrial adenosine triphosphate sensitive potassium (K(ATP)) channel, a major contributor to ischemic cardioprotection. METHODS: Fifty patients undergoing angioplasty of a major epicardial coronary artery were randomly allocated to either ischemic preconditioning or intracoronary estrogen administration in the presence or absence of glibenclamide (glyburide). RESULTS: The coronary collateral circulation, as quantitatively assessed by an intracoronary Doppler flow wire, was similar during balloon inflation among the groups. Patients in the preconditioned and estrogen-treated groups significantly lowered their ischemic burden, as assessed by an ST-segment shift, chest pain score and myocardial lactate extraction ratio, as compared with control subjects. The reduction in the ST-segment shift afforded by estrogen during the first inflation (-63% vs. first inflation in the preconditioned group) was similar to that afforded by preconditioning during the second inflation (-68% vs. first inflation). In contrast, the patients given glibenclamide developed significantly higher ischemic burden during the first and second inflations, as compared with those in the estrogen-treated group alone. CONCLUSIONS: It is concluded that intracoronary administration of estrogen before balloon angioplasty rendered the myocardium relatively resistant to subsequent ischemia, and the degree of cardioprotective effect was comparable to that afforded by ischemic preconditioning. The effect of estrogen was abolished by glibenclamide, suggesting that the cardioprotective effect of estrogen may result from activation of myocardial K(ATP) channels. PMID- 11869856 TI - Risk of cardiovascular disease in relation to achieved office and ambulatory blood pressure control in treated hypertensive subjects. AB - OBJECTIVE: We investigated the prognostic impact of 24-h blood pressure control in treated hypertensive subjects. BACKGROUND: There is growing evidence that ambulatory blood pressure improves risk stratification in untreated subjects with essential hypertension. Surprisingly, little is known on the prognostic value of this procedure in treated subjects. METHODS: Diagnostic procedures including 24-h noninvasive ambulatory blood pressure monitoring were undertaken in 790 subjects with essential hypertension (mean age 48 years) before therapy and after an average follow-up of 3.7 years (2,891 patient-years). RESULTS: At the follow-up visit, 26.6% of subjects achieved adequate office blood pressure control (<140/90 mm Hg), and 37.3% of subjects achieved adequate ambulatory blood pressure control (daytime blood pressure <135/85 mm Hg). Months or years after the follow-up visit, 58 patients suffered a first cardiovascular event. Event rate was lower (0.71 events/100 person-years) among the subjects with adequate ambulatory blood pressure control than among those with higher blood pressure levels (1.87 events/100 person-years) (p = 0.0026). Ambulatory blood pressure control predicted a lesser risk for subsequent cardiovascular disease independently of other individual risk factors (RR 0.36; 95% confidence intervals: 0.18 to 0.70; p = 0.003), including age, diabetes and left ventricular hypertrophy. Office blood pressure control was associated with a nonsignificant lesser risk of subsequent events (RR 0.63; 95% confidence intervals: 0.31 to 1.31; p = NS). In-treatment ambulatory blood pressure was more potent than pre-treatment blood pressure for prediction of subsequent cardiovascular disease. CONCLUSIONS: Ambulatory blood pressure control is superior to office blood pressure control for prediction of individual cardiovascular risk in treated hypertensive subjects. PMID- 11869857 TI - A prognostic model for predicting the disappearance of left atrial thrombi among candidates for percutaneous transvenous mitral commissurotomy. AB - OBJECTIVES: We sought to develop a prognostic model to predict the disappearance of left atrial thrombi (LAT) among candidates for percutaneous transvenous mitral commissurotomy (PTMC). BACKGROUND: Complete LAT resolution can be achieved with oral anticoagulation, allowing a number of patients to safely undergo PTMC. METHODS: We randomly allocated 108 PTMC candidates with LAT into two subsets-- one to derive the model and the other to validate it. The existence of LAT and its size were measured by transesophageal echocardiography. Patients were given oral anticoagulation and followed up for 6 to 34 months. There was a 62% disappearance rate of LAT. RESULTS: We developed the following model: P = 1/(1 + exponential [-8.1 + 1.8 NYHA + 0.7 area]), where NYHA = New York Heart Association functional class (from I to IV), and area = LAT area (in cm(2)). The model was well calibrated (goodness-of-fit test, p = 0.82) and well discriminated (area under the receiver-operating characteristics [ROC] curve = 0.92). Performance in the validating sample was equally good (area under the ROC curve = 0.94; goodness-of-fit test, p = 0.16). When a cut-off point of p > 0.7 was used to designate the LAT disappearance in the validating set, the model had a sensitivity, specificity and positive and negative predictive values of 93.3%, 79.2%, 84.9% and 90.5%, respectively. CONCLUSIONS: Combined clinical (NYHA functional class) and echocardiographic (LAT area) variables are predictive of the 34-month outcome of oral anticoagulation for LAT resolution among PTMC candidates. This simple and highly predictive model might be potentially useful for clinical assessment and proper management. PMID- 11869858 TI - The detection of cardiotropic viruses in the myocardium of patients with arrhythmogenic right ventricular dysplasia/cardiomyopathy. AB - OBJECTIVES: We sought to investigate the role of cardiotropic viruses, including adenovirus, cytomegalovirus (CMV), enterovirus and parvovirus, in arrhythmogenic right ventricular dysplasia/cardiomyopathy (ARVD/C). BACKGROUND: Arrhythmogenic right ventricular dysplasia/cardiomyopathy is characterized by a gradual loss of myocytes, inflammatory infiltrates and replacement by fatty and fibrous tissue. It has been speculated that ARVD/C is a sequela of viral myocarditis in some patients, and the role of the coxsackievirus B3 has been debated. METHODS: Myocardial samples from 12 patients with ARVD/C were analyzed by polymerase chain reaction for the presence of cardiotropic viruses. RESULTS: Enteroviral sequences were detected in seven patients and adenovirus type 5 in another two patients. During the same period, 215 control samples were analyzed in which only CMV (n = 2) and enterovirus (n = 1) were detected. This suggests a link between ARVD/C and the presence of viral genome (enterovirus or adenovirus) in the myocardium. CONCLUSIONS: We report that cardiotropic viruses are more frequently identified in patients with ARVD/C than in control subjects. However, the role of these viruses in ARVD/C pathogenesis remains unknown. PMID- 11869859 TI - Sensitivity and specificity of concealed entrainment for the identification of a critical isthmus in the atrium: relationship to rate, anatomic location and antidromic penetration. AB - OBJECTIVES: This study was designed to determine the sensitivity and specificity of concealed entrainment (CE) for the identification of a critical isthmus in the atrium. BACKGROUND: Isthmus identification during entrainment mapping of macro reentrant atrial tachycardia (MRAT) relies on the demonstration of CE. METHODS: Using the model of typical atrial flutter, entrainment was performed in 10 patients at four rates (flutter cycle length [FCL] minus 10/20/30/40 ms) from seven sites: isthmus entrance/exit, low lateral/high lateral/high septal right atrium and proximal/distal coronary sinus. Surface 12-lead electrocardiogram fusion was evaluated by three observers blind to patient status. The extent of antidromic penetration (AP) was measured off the pacing catheter positioned around the tricuspid annulus. RESULTS: The sensitivity for CE identifying any isthmus site was greatest at FCL-10 (100%), but the specificity was poor (54%). Conversely, specificity was greatest at FCL-40 (98%), but the sensitivity was poor (65%), with manifest entrainment (ME) observed from the isthmus entrance in 70% of episodes. At FCL-30, sensitivity (85%) and specificity (90%) were "balanced," but CE still resulted during entrainment from a non-isthmus site in five of 10 patients. Antidromic penetration increased with pacing CL shortening (p < 0.001) and correlated with the development of ME (p < 0.001). Antidromic penetration was significantly blunted from the isthmus exit compared to all other sites (p = 0.003). CONCLUSIONS: The sensitivity and specificity of CE for identifying an isthmus in the atrium are critically dependent on the pacing rate and the precise anatomic pacing site within the isthmus. These findings may have implications for the use of entrainment in the mapping of unknown MRAT circuits. PMID- 11869860 TI - The antioxidant N-2-mercaptopropionyl glycine attenuates left ventricular hypertrophy in in vivo murine pressure-overload model. AB - OBJECTIVES: In order to identify the role of reactive oxygen species (ROS) in cardiac hypertrophy, we examined the effect of N-2-mercaptopropionyl glycine (MPG) on cardiac hypertrophy. BACKGROUND: Recent in vitro studies have suggested that ROS play an important role as a second messenger in cardiac hypertrophy. It was therefore thought to be of particular value to examine the relevance of studies using in vitro models for cardiac hypertrophy in an in vivo setting. METHODS: The transverse thoracic aorta in mice was constricted, and MPG (100 mg/kg) was infused intraperitoneally twice daily. The animals were assessed seven days after the operation for hemodynamic functions, oxidative stress and antioxidative enzyme activities. RESULTS: Banding of the transverse aorta in mice resulted in an increase in the ratio of heart weight to tibia length and the appearance of an endogenous atrial natriuretic factor messenger ribonucleic acid (mRNA) seven days postoperatively. Administration of MPG significantly attenuated the hypertrophic responses induced by pressure overload. Cardiac hypertrophy was accompanied by increases in heme oxygenase-1 mRNA expression and lipid peroxidation, which was eliminated by the treatment with MPG. Pressure overload led to increases in antioxidant enzyme activities, such as superoxide dismutase and glutathione peroxidase, but not catalase, activity. CONCLUSIONS: Our results indicated that oxidative stress was increased in our model and that it plays an important role in the development of cardiac hypertrophy. PMID- 11869861 TI - Top line mentality. PMID- 11869862 TI - President's page: mission possible: with help from our friends. PMID- 11869864 TI - Double potentials analysis as a guide to radiofrequency ablation of atrial flutter. PMID- 11869865 TI - Decreased incidence of postoperative stroke following off-pump coronary artery bypass. PMID- 11869867 TI - Cytokine gene polymorphisms and development of CAD associated with CP infection. PMID- 11869868 TI - Patients with CHF and depression have greater risk of mortality and morbidity than patients without depression. PMID- 11869871 TI - Inhibition of cell differentiation by manganese chloride in micromass cultures of mouse embryonic limb bud cells. AB - Although manganese is an essential element, in excess it has proven to be toxic to a number of organ systems. In several studies, exposure to manganese compounds during embryological development has been linked to skeletal abnormalities. Micromass cultures of dissociated mouse embryonic limb bud cells were used to investigate whether manganese could interfere with the process of chondrogenesis. Cell viability was significantly reduced in cultures grown in the presence of 1 and 10 mM MnCl2, whereas as little as 10 microM MnCl2 inhibited differentiation of limb cells into proteoglycan-producing nodules. Because the concentration of MnCl2 needed to interfere with differentiation is so much lower than the cytotoxic concentration, it is suggested that manganese has the potential to be teratogenic, and to specifically interfere with the development of a cartilage matrix, although by an unknown mechanism. PMID- 11869872 TI - Effects of Mannich bases on cellular glutathione and related enzymes of Jurkat cells in culture conditions. AB - Jurkat cells were exposed to representative acetophenone-derived mono Mannich bases 2 and 3 and also cyclic Mannich base C1 in culture conditions to see the alterations in the most abundant cellular thiol, glutathione and also some of the enzymes in its metabolic pathway. Jurkat cells were exposed to the compounds for 24 h in cell culture medium with fetal bovine serum (1%) at 37 degrees C under a humidified atmosphere of 95% air and 5% CO2. Mannich bases generally increased total glutathione level (123-151% of control). Glutathione S-transferase (GST) activity also increased (150-363% of control), while glutathione disulfide reductase (GRD) activity was not affected. The increase in cellular glutathione level may possibly result from de novo glutathione synthesis. The consumption of the glutathione due to alkylation by Mannich bases might have stimulated the enzymes in the gamma-glutamyl cycle in our experimental design, where the cells had nutrients and time to react with their feedback mechanisms. A remarkable increase in GST activity might be a compensatory up-regulation to detoxify Mannich bases by conjugating them with cellular thiols. PMID- 11869874 TI - Effect of UV-B radiation on some common antibiotics. AB - Some of the commonly used antibiotics such as cephaloridine, cephalexin, cephradine, nystatin and nafcillin were tested for generation of singlet oxygen (1O(2)) under UV-B (290-320 nm) exposure and the order for 1O(2) generation was obtained: cephaloridine>cephalexin>nystatin>cephradine>nafcillin. In vitro study with deoxyguanosine (dGuo) showed that 1O(2) was responsible for drug-sensitized photodegradation of the guanine base of DNA and RNA. Sodium azide (NaN(3)) and 1,4-diazabicyclo [2.2.2] octane (DABCO) accorded significant inhibition (76-98%) in the production of (1)O(2) and photo-oxidation of dGuo. The combined effect of drug and UV-B irradiation is of paramount importance in view of cell-damaging reactions by 1O(2). Our findings are important because of increasing UV-B radiation on the earth's surface due to depletion of the stratospheric ozone layer. The selected drugs are used routinely for the treatment of various diseases and their combined action may cause undesirable phototoxic responses. Our study suggests that exposure to sunlight should be avoided after the intake of the photosensitive drugs. PMID- 11869873 TI - Effects of 3-MeSO2-DDE and some CYP inhibitors on glucocorticoid steroidogenesis in the H295R human adrenocortical carcinoma cell line. AB - The formation of steroids in the H295R human adrenocortical carcinoma cell line was analysed by HPLC or RIA, and based on these data the apparent catalytic activities of CYP11A, CYP17, CYP21 and CYP11B1 in this cell line were calculated. The environmental pollutant 3-methylsulfonyl-DDE (3-MeSO2-DDE) and the cytochrome P450 (CYP) inhibitors ketoconazole, metyrapone and aminoglutethimide were studied for their effects on the steroid formation. Metyrapone (IC50) of 1 microM) and 3 MeSO2-DDE (10 microM: 66 +/- 10% of control) were found to inhibit the apparent CYP11B1 activity. Ketoconazole inhibited all enzymes examined with the greatest effects on CYP11B1 (IC50) of 2.5 microM). Aminoglutethimide was examined only for effects on CYP11A activity and was shown to inhibit pregnenolone formation (20 microM: 61 +/- 4% of control). The possibility of studying all CYP enzymes in the corticosteroidogenesis makes this cell line a valuable test system to examine effects of chemicals, such as suspected endocrine disruptors, on the human glucocorticoid hormone synthesis. The inhibition of cortisol formation by 3-MeSO2 DDE supports an interaction with the active site of CYP11B1, as previously reported in mouse adrenocortical Y1 cells. In mice, this interaction led to metabolic activation and a high adrenotoxicity of 3-MeSO2-DDE. Therefore studies on the adrenotoxicity of 3-MeSO2-DDE in humans are needed. PMID- 11869875 TI - Influence of tris(4-chlorophenyl)methanol (TCPM) on gap junction-mediated intercellular communication of cultured bovine granulosa cells. AB - Tris(4-chlorophenyl)methanol (TCPM) is a by-product in the manufacture of technical grade DDT, which is known to alter properties and functions of the female reproductive system. We investigated whether in vitro TCPM has an influence on the function of gap junction-mediated intercellular communication (GJIC) and gap junction protein expression of connexin 43 (Cx43) in cultured bovine granulosa cells. GJIC was assessed by fluorescent dye microinjection (dye coupling). After a 1-h exposure to TCPM at a concentration of 32 microM, a significant (P<0.05) reduction in dye coupling occurred. The same result was obtained with o,p'-DDT. At a concentration of 32 microM both pesticides were cytotoxic as indicated by significant (P<0.05) increased propidium iodide staining of the cell nuclei. Little or no effect on the stainable pattern of connexons occurred after 1 h incubation time, while after 3 h treatment from 16 to 64 microM TCPM, a significant inhibition in the immunostaining resulted and the concentrations of 32 and 64 microM TCPM were cytotoxic for the granulosa cells. The freeze-fracture electron microscopy resulted in small differences in the morphology of gap junction plaques of cell cultures treated for 3 h with 8 or 16 microM TCPM in comparison to untreated cells. After treatment with 32 microM TCPM, gap junction plaques were very rarely detected and the lateral intramembraneous particles (IMP) distribution of many plasma membranes was strongly altered. Estimation of the cellular parameters may lead to an enhanced understanding of the mechanism of chemically induced toxicity by TCPM, that causes a general toxic effect on granulosa cells. We can conclude that TCPM is a toxic risk in the same manner as DDT. PMID- 11869876 TI - Inhibitory effects of organotin compounds on voltage-dependent, tetrodotoxin resistant Na+ channel current in guinea pig dorsal root ganglion cells. AB - The effects of organotin compounds on voltage-dependent, tetrodotoxin (TTX) resistant Na+ channel current (I(Na)) in single cells isolated from guinea pig dorsal root ganglion were investigated using a whole cell patch clamp technique. Extracellular application of tributyltin (TBT) inhibited I(Na) in a concentration dependent manner with an IC50 of 7.2 microM. TBT (100 microM), when applied intracellularly, was without effect. Triphenyltin (TPT, 100 microM) and dibutyltin (DBT, 100 microM), applied extracellularly, inhibited I(Na) with an efficacy ranking of TBT>TPT>DBT. Monobutyltin (100 microM), whether applied externally or internally, had little effect on I(Na). TBT (30 microM) significantly prolonged both time to peak and half-decay time of I(Na) and shifted the activation curve of I(Na) in the positive direction without changing the slope. No such effect was produced by TPT (100 microM). The results indicate that organotin compounds inhibit voltage-dependent, TTX-resistant Na+ channel activity and suggest that the inhibitory action may account, at least in part, for their neurotoxic effects. PMID- 11869877 TI - Toxicity of ethacrynic acid in isolated rat hepatocytes. AB - Ethacrynic acid, a loop diuretic drug, caused lipid peroxidation in isolated rat hepatocytes. The thiobarbituric acid reactive substances (TBARS) formation showed a good correlation with the leakage of glutamic-oxaloacetic acid transaminase (GOT) from the hepatocytes. The addition of antioxidants such as N, N'-diphenyl-p phenylenediamine (DPPD) and promethazine to the isolated rat hepatocyte suspension containing ethacrynic acid prevented the lipid peroxidation and decreased the GOT leakage to some extent. SKF-525A inhibited the oxidative metabolism of ethacrynic acid and decreased the TBARS formation, suggesting that the lipid peroxidation was caused by the oxidative metabolism. The intracellular reduced glutathione markedly decreased in the hepatocyte suspension containing ethacrynic acid and the hepatocellular protein sulfhydryls were decreased, which was negatively correlated with the GOT leakage. Thus the ethacrynic acid-induced hepatotoxicity was found to be related to the lipid peroxidation and the decrease of cellular protein sulfhydryls. PMID- 11869878 TI - Effects of the ionophores valinomycin, ionomycin and gramicidin A on the element compartmentation in cultured rat hepatocytes. AB - The element compartmentation in cultured rat hepatocytes was studied by electron probe X-ray microanalysis of freeze-dried cryosections after exposure of the cells to the ionophores valinomycin, ionomycin or gramicidin A. The most striking effect of these ionophores is the decrease of the intracellular potassium/sodium ratio from values of approximately 10 under control conditions to values below 1 after application of the ionophores. Changes of sodium, potassium and chloride are similar in cytoplasm and nucleus. However, elemental changes are delayed or impeded in mitochondria with respect to the surrounding cytoplasm. The water portion of cytoplasm and mitochondria slightly increases. Besides that, each ionophore has specific effects on the intracellular ion distribution. As compared to gramicidin A and ionomycin, valinomycin does not change the intracellular chloride content. Ionomycin induces calcium accumulation in mitochondria. The cytotoxic effects of the studied ionophores on the intracellular element distribution are more complex than supposed from their ion selective properties in membranes. PMID- 11869879 TI - Chronic exposure of HepG2 cells to excess copper results in depletion of glutathione and induction of metallothionein. AB - Metallothionein (MT) and reduced glutathione (GSH) play an important role in the intracellular handling of copper by preventing the generation and favouring the removal of copper-derived free radicals. The present study addressed the changes in MT and GSH that follow chronic (2 or 5 weeks) exposure of human hepatoblastoma cells (HepG2) to excess copper. Copper treatment (100 microM, 2 weeks) led to a 28-fold elevation in intracellular copper. Concomitantly, cells exhibited a seven fold increase in total MT and an increment in its saturation with copper from 45 to 86%. Around 38% of copper in the cytosolic fraction could be accounted for by MT. GSH equivalents were substantially lowered (to 37% of basal levels) in treated cells, with only part of it being accounted for by an increase in GSSG. Copper-treatment induced no changes in catalase or GSH-peroxidase activities but it was associated with a small reduction in SOD (20%) and GSH-reductase (26%) activities. Copper-loaded cells did not differ from controls in their basal oxidative tone; however, when exposed to tert-butylhydroperoxide they exhibited a markedly greater susceptibility to undergo both oxidative stress and cell lysis. It is proposed that chronic exposure of HepG2 cells to excess copper is accompanied by "adaptive changes" in GSH and MT metabolism that would render cells substantially more susceptibility to undergo oxidative stress-related cytotoxicity. PMID- 11869880 TI - In vitro application of the comet assay for aquatic genotoxicity: considering a primary culture versus a cell line. AB - The comet assay, one of the most widely used techniques for the evaluation and detection of DNA strand breaks, is frequently employed in vivo. In vitro assays are usually performed with mammalian cell lines, clearly not the best choice for tests on aquatic genotoxicity. Here we evaluated a fish hepatoma cell line (RTH 149) and a primary blood cell culture from the intertidal colonial tunicate Botryllus schlosseri as possible model targets for comet assays using the genotoxic agent H2O2. We found that DNA strand break levels in RTH-149 fitted dose-dependent responses better than the tunicate cells. Moreover, in B. schlosseri controls, 34% of the cells were already ranked as severely damaged. Assays were then performed on water samples from the polluted Kishon river (Israel) on three different dates, using RTH-149 cells (50% dilutions, 2-h exposures). In all cases, high genotoxicity of the river water was revealed by evaluating comet percentages, average tail lengths and DNA damage levels. This assay was found to be fast and sensitive, appropriate to be employed as a part of a monitoring program. The use of B. schlosseri blood cells should be validated in additional work. PMID- 11869881 TI - Exposure of human lung cells to native diesel motor exhaust--development of an optimized in vitro test strategy. AB - To investigate the effects of native diesel motor exhaust on human lung cells in vitro, a new experimental concept was developed using an exposure device on the base of the cell cultivation system CULTEX (Patent No. DE19801763.PCT/EP99/00295) to handle the cells during a 1-h exposure period independent of an incubator and next to an engine test rig. The final experimental set-up allows the investigation of native (chemically and physically unmodified) diesel exhaust using short distances for the transportation of the gas to the target cells. The analysis of several atmospheric compounds as well as the particle concentration of the exhaust was performed by online monitoring in parallel. To validate the complete system we concentrated on the measurement of two distinct viability parameters after exposure to air and undiluted, diluted and filtered diesel motor exhaust generated under different engine operating conditions. Cell viability was not influenced by the exposure to clean air, whereas dose-dependent cytotoxicity was found contingent on the dosage of exhaust. Additionally, the quality of exhaust, represented by two engine operating conditions (idling, higher load), also showed well-distinguishable cytotoxicity. In summary, the experimental set up allows research on biological effects of native engine emissions using short exposure times. PMID- 11869882 TI - Evaluation of the cytotoxicity of selected systemic and intravitreally dosed drugs in the cultures of human retinal pigment epithelial cell line and of pig primary retinal pigment epithelial cells. AB - The cytotoxicity of the selected systemic and intravitreally dosed drugs tamoxifen, toremifene, chloroquine, 5-fluorouracil, gentamicin and ganciclovir was studied in retinal pigment epithelium (RPE) in vitro. The cytotoxicity was assayed in the human RPE cell line D407 and the pig RPE cell culture using the WST-1 test, which is an assay of cell proliferation and viability. The effects of experimental conditions on the WST-1 test (cell density, serum content in the culture medium, the exposure time) were evaluated. The EC50 values in tamoxifen treated D407 cells ranged between 6.7 and 8.9 micromol/l, and in pig RPE cells between 10.1 and 12.2 micromol/l, depending on the cell density used. The corresponding values for toremifene were 7.4 to 11.1 micromol/l in D407 cells and 10.0 to 11.6 micromol/l in pig RPE cells. In chloroquine-treated cells, the EC50 values were 110.0 micromol/l for D407 cells and 58.4 micromol/l for pig RPE cells. Gentamicin and ganciclovir did not show any toxicity in micromolar concentrations. The exposure time was a significant factor, especially when the drug did not induce cell death, but was antiproliferative (5-fluorouracil). Serum protected the cells from the toxic effects of the drugs. Both cell cultures were most sensitive to tamoxifen and toremifene, and next to chloroquine. The drug toxicities obtained in the present study were quite similar in both cell types; that is, the pig RPE cells and the human D 407 cell line, despite the differences in, for example, the growth rate and melanin contents of the cell types. Owing to the homeostatic functions important for the whole neuroretina, RPE is an interesting in vitro model for the evaluation of retinal toxicity, but, in addition to the WST-1 test, more specific tests and markers based on the homeostatic functions of the RPE are needed. PMID- 11869883 TI - A transformed fish cell line expressing a green fluorescent protein-luciferase fusion gene responding to cellular stress. AB - We obtained a stable transformed fish (EPC) cell line containing a reporter gene under the control of the tilapia HSP70 promoter. Expression of the reporter gene, coding for a green fluorescent protein (GFP)-luciferase fusion protein, was assessed by measuring the luciferase enzymatic activity by luminometry and the GFP expression by fluorescence microscopy and flow cytometry. The clone was characterized for its capacity to respond to heat shock treatment. The results show high induction after 1 h at 37 degrees C of treatment, up to 500-fold. In addition, its convenience to detect a large range of cellular stressors was evaluated. We observed high induction when Cd2+, Zn2+, Hg2+ or Cu2+ was added, but not Pb2+. In addition, activation of the reporter gene was observed in the presence of other compounds such as acetyl chloride, tetrachlorophenol, chloroacetamide and sodium arsenite. In conclusion, this cell line can be used as a rapid, cheap and easy biological test to determine cellular stress induced by environmental pollutants, alone or in conjunction with other, more specific assays. PMID- 11869888 TI - Prophylactic cancer vaccines. AB - Increasingly, data from distinct experimental systems show that immunity can be activated to prevent tumors. The rationale for prevention is strong because, in that setting, one deals with an immune system that is neither impaired by tumor- and treatment-induced suppression nor tolerant to tumor-associated antigens that have been encountered in the absence of correct presentation and costimulatory/danger signals. The use of overexpressed or mutated proteins, or mutated oncogenic growth factor receptors, as tumor-associated antigens yields rational targets for specific immunoprevention. Transgenic mouse models are providing encouraging indications of future usefulness of vaccines that are based on these molecules. PMID- 11869887 TI - NKT cells - conductors of tumor immunity? AB - NKT cells are key players in the regulation of antitumor immunity, particularly in experimental models of tumor immunotherapy, such as IL-12 or alpha galactosylceramide administration. They may also operate in natural antitumor immunity. NKT cells are best known for their immunosuppressive functions; however, NKT cells interact with a range of other cell types (particularly dendritic cells and NK cells) and the outcome of NKT-cell stimulation depends on these and on the cytokine/co-stimulatory milieu. PMID- 11869889 TI - Clinical cancer vaccine trials. AB - Antigens that are selectively or abundantly expressed in cancer cells have been used for clinical trials, mostly in patients with advanced disease, and appear to be better vaccines than whole cells. Candidate vaccines have emerged from different categories of cancer antigens. Strategies involving various forms of peptides have been used either alone or combined with different cytokines, adjuvants or dendritic cells to enhance specific immune responses. Although individual patients have benefited, no strategy has emerged as universally applicable; neither has any route of administration. Increasingly sensitive methods have correlated clinical responses with measurable immune responses to vaccination in some patients. PMID- 11869891 TI - Ontogeny and genetics of the hemato/lymphopoietic system. AB - During embryogenesis there is a sequential, temporal appearance of increasingly more-complex hematopoietic cells beginning with unipotential progenitors, proceeding to multipotential (myeloid, erythroid and lymphoid) progenitors and culminating with adult-repopulating hematopoietic stem cells. Current research has established an important role for the aorta-gonads-mesonephros region of the mouse embryo in the generation of multipotential progenitors and hematopoietic stem cells. Comparisons of normal and hematopoietic-cell-mutant mouse embryos have revealed several genes pivotal in hematopoietic stem cell generation/function. Other genes have been implicated in the critical generation of lymphoid lineage potential. Thus, an understanding of the cellular and molecular interactions within the midgestation aorta-gonads-mesonephros region offers insight into the mechanisms of hematopoietic lineage specification during ontogeny and perhaps will lead to a more complete knowledge of the adult hematopoietic system. PMID- 11869892 TI - Deciphering the role of Notch signaling in lymphopoiesis. AB - Components of the Notch signaling pathway are expressed during multiple stages of lymphoid development. Consistent with its function during invertebrate development, Notch signaling is proposed to have a central role in lymphoid cell fate specification. Recent studies show that Notch signaling is a proximal event in T-cell commitment from a common lymphoid progenitor. The role of Notch at later stages of lymphoid development is controversial, but recent data suggest models that may help clarify observations. Current studies suggest that Notch activity is cell-context dependent and interactions between Notch and other environmental receptors are integrated during cell-fate decisions. Furthermore, the requirement for precise regulation of Notch activity is evident from human and murine neoplasms in which dysregulated Notch signaling leads to T-cell leukemia. Future studies that identify the stages of lymphoid development where Notch signaling is physiologically active and the exact targets of Notch signaling that are relevant to lymphopoiesis should significantly improve our understanding of Notch function in T- and B-cell development. PMID- 11869893 TI - On the brink of becoming a T cell. AB - Recent studies provide fresh insight into the mechanisms by which precursor cells are committed to and develop within the T-lymphocyte lineage. Precursor/product studies have identified developmental stages between that of the pluripotent hematopoietic stem cell and thymocytes committed to the T lineage. Specific ligands and signaling pathways interacting with the Notch-1 receptor and its ability to influence commitment within the lymphoid lineage have been described. Although the structural features or putative ligands endowing the pre-TCR with constitutive signaling capacity remain elusive, numerous distal mediators of pre TCR signaling have been identified. It remains for the future to determine what roles they may have in survival, proliferation, lineage commitment and allelic exclusion of TCR genes. Receptor editing and lineage commitment of alphabeta T cells still represent controversial topics that need further study. PMID- 11869894 TI - New perspectives on a developmental dilemma: the kinetic signaling model and the importance of signal duration for the CD4/CD8 lineage decision. AB - Double-positive thymocytes are short-lived bipotential cells whose developmental fate is determined by the specificity of their TCRs. A relatively small number of double-positive thymocytes undergo positive selection in the thymus and these are signaled to differentiate either into CD4(+) or CD8(+) mature T cells. The mechanism by which double-positive thymocytes determine their appropriate CD4/CD8 fate has been the subject of intense theoretical debate and rigorous experimental analysis. In the last year, 'signal duration' has been offered as a replacement for 'signal strength' as a major determinant of the CD4/CD8 decision, a deceptively minor refinement that requires a major change in our understanding of how signaled double-positive thymocytes differentiate into mature T cells. Indeed, the kinetic signaling model provides a radically new perspective on the mechanism by which the CD4/CD8 lineage decision is made. PMID- 11869895 TI - Transcriptional control of B-cell development. AB - Significant progress has recently been made in our understanding of how transcription factors such as PU.1, Notch1, E2A, EBF, Pax5, Bcl6, Blimp1 and XBP1 control different developmental decisions during the onset and terminal phase of B-lymphopoiesis. One emerging theme is that negative regulatory networks play an important role in suppressing alternative gene programs and their corresponding cell fates throughout B-cell development. PMID- 11869896 TI - A recombinase diversified: new functions of the RAG proteins. AB - The RAG proteins were long thought to serve merely as a nuclease, initiating recombination by cleaving DNA. Recent work has shown, however, that these proteins are essential for many steps in the recombination pathway, such as opening hairpins and joining broken DNA ends, and that they can also act as a transposase, targeting distorted DNA structures such as hairpins. PMID- 11869897 TI - Allelic exclusion at the TCRbeta locus. AB - Assembly of TCRbeta chain variable-region genes is regulated in the context of allelic exclusion. Differential epigenetic modifications of the two TCRbeta alleles established early in embryonic development may be important for permitting allelic exclusion by ordering rearrangement of the two alleles in double-negative thymocytes. Expression of a TCRbeta chain, as part of the pre-TCR complex, activates signaling pathways that enforce allelic exclusion in double positive thymocytes. These signaling pathways, which utilize p56(lck) and SLP-76, may be distinct from those used to promote other processes initiated by pre-TCR expression. In double-positive thymocytes allelic exclusion is enforced, in part, by changes in Vbeta gene segment accessibility promoted by cis-acting elements that may be distinct from those regulating accessibility of D/Jbeta gene segments. PMID- 11869899 TI - Antigen-dependent B-cell development. AB - There is growing evidence that the development of naive B cells depends on the interaction of self antigens with the BCR. A view that has emerged over the past year is that BCR signal output contributes in a large part to the developmental fate of peripheral B cells. Differences in antigen-receptor signal strength may determine whether B cells assume a marginal zone, follicular or B-1 phenotype. PMID- 11869900 TI - Development and selection of NKT cells. AB - NKT cells utilize a restricted alphabeta TCR repertoire that recognizes glycolipids in association with CD1d. The recent development of fluorescent CD1d tetramers loaded with the synthetic glycolipid alpha-galactosyl-ceramide has led to a clearer definition of NKT-cell subsets as well as important insights into their developmental origin. As many as four subsets may exist, differing in NK1.1 expression, TCR repertoire and dependence on CD1d and various glycolipids for development. Two different lineage-commitment models have been proposed, with most evidence favoring a byproduct of conventional-T-cell development. PMID- 11869898 TI - Somatic immunoglobulin hypermutation. AB - Immunoglobulin hypermutation provides the structural correlate for the affinity maturation of the antibody response. Characteristic modalities of this mechanism include a preponderance of point-mutations with prevalence of transitions over transversions, and the mutational hotspot RGYW sequence. Recent evidence suggests a mechanism whereby DNA-breaks induce error-prone DNA synthesis in immunoglobulin V(D)J regions by error-prone DNA polymerases. The nature of the targeting mechanism and the trans-factors effecting such breaks and their repair remain to be determined. PMID- 11869901 TI - Gut intraepithelial lymphocyte development. AB - CD8alphabeta(+) and CD4(+) intraepithelial lymphocytes, the progeny of double positive thymocytes, are oligoclonal T-cell populations that have accumulated in the gut wall as the result of repeated antigenic stimulations, which lead to rounds of traffic through the lymph/blood circuit ending in an alpha4beta7 integrin-driven homing all along the gut mucosa. In contrast, CD8alphaalpha(+) intraepithelial lymphocytes, which may be TCRgammadelta(+) or alphabeta(+), result in part from local differentiation in the gut, but studies comparing euthymic and athymic mice suggest a thymic double-negative origin for many of them. PMID- 11869902 TI - Novel approaches for identifying genes regulating lymphocyte development and function. AB - The draft sequence of the human and mouse genomes provides an unparalleled opportunity for understanding the genetic control of immune-cell development. Strategies can begin with a gene sequence and pursue a putative immune-system function by employing mRNA-expression profiling or creating gene knockouts in embryonic stem cells. The latter can be produced by utilising the Cre/Lox system, a tetracycline operon, a gene-trap method or chemical mutagenesis. Alternatively, mutant phenotypes (derived using the mutagen ethylnitrosourea) can be traced back to gene sequences. PMID- 11869903 TI - BAFFled B cells survive and thrive: roles of BAFF in B-cell development. AB - Interactions of BAFF (B-cell activating factor) with BAFF-R, one of three BAFF binding receptors that are preferentially expressed on B cells, are essential for B-cell development, because defects in either the ligand or the receptor arrest progression from immature type-1 B cells to type-2 cells and mature cells; B1 B cells are unaffected. Transgenic BAFF overexpression leads to B-cell hyperplasia and autoimmune disease. In vitro, BAFF increases survival of immature and mature B cells; immature B cells also mature polyclonally to mature B cells, without proliferation. Upon BAFF-influenced differentiation, immature B cells change their surface-IgM signal transduction machinery and proliferate rather than undergoing apoptosis. PMID- 11869904 TI - A method for determination of upper extremity kinematics. AB - Kinematic analysis of the upper extremity has been conducted using a wide variety of techniques, philosophies, and analytic methods. We describe a simple, marker based three-dimensional video analytic technique that borrows concepts from lower extremity kinematic analysis. A sequential rotation order about orthogonal axes is described, although alternate methods are examined as well. The method has been verified by application to a mechanical model. In certain positions, gimbal lock may occur, and a different sequence of rotational decomposition may be required. Agreement on standardization of technique would assist in the dissemination of upper extremity scientific data. PMID- 11869906 TI - Length and force of the gastrocnemius and soleus during gait following tendo Achilles lengthenings in children with equinus. AB - Nine subjects (12 sides) with cerebral palsy who walked in equnius were evaluated prior to and 1 year after surgical tendo Achilles lengthening. Gastrocnemius and soleus length [Gait Posture, 6 (1997) 9] and plantarflexor force [Gait Posture, 6 (1997) 9; J Biomech, 23 (1990) 495] were calculated. The length of the gastrocnemius and soleus increased significantly (P<0.01) following the intervention. Force output of the triceps surae during push-off increased significantly (13.95 N/kg body weight (BW) preop to 30.31 N/kg BW postop; P<0.01). Assessment of the force-length capacity of the triceps surae in candidates for tendo Achilles lengthenings may identify individuals at risk of residual weakness and iatrogenic crouch. PMID- 11869905 TI - Upper body movement during walking in children with lumbo-sacral myelomeningocele. AB - Eight children with lumbo-sacral myelomeningocele (MMC) underwent three dimensional movement analysis to determine whether or not differing levels of lower extremity strength affected the extent of shoulder, trunk and pelvis movement during independent walking when wearing orthoses. Fourteen control children were also investigated. The patterns of upper body movements in all MMC children were well defined and consistent, showing small standard deviations from the mean. In the frontal and transverse planes, segment displacements of the MMC children assigned into Group II (hip extensor and abductor muscle strength grade 0-2) were almost twice that of the MMC children in Group I (hip extensor and abductor muscle strength grade 3-4). All segment displacements in the frontal, transverse and sagittal planes for Group I and Group II children were significantly greater than those for the controls. In the frontal plane these differences were approximately 4-10 times greater, with the Group II children having the largest peak-to-peak displacements. These results indicate that the motion amplitudes of the upper body segments are related to the degree of muscle weakness of the lower limbs. No significant differences were found when comparing segment motions during walking with either the Ferrari type knee-ankle-foot or ankle-foot orthoses. PMID- 11869907 TI - Intra-subject repeatability of the three dimensional angular kinematics within the lumbo-pelvic-hip complex during running. AB - The intra-subject repeatability of measuring the three dimensional (3D) angular kinematics of the lumbo-pelvic-hip complex during running was evaluated. Spatio temporal parameters were found to be repeatable. All angular kinematic parameters displayed high within-day repeatability despite numerous potential sources of variability in the data. Most angular kinematic parameters also displayed high between-day and overall repeatability, except for trunk flexion-extension, lumbar spine flexion-extension, pelvic anterior-posterior tilt, hip axial rotation and thigh axial rotation. These angular parameters were highly susceptible to marker reapplication errors that produced static daily offsets in the data. Overall, the results of this study suggest that the 3D angular kinematic patterns of the lumbo pelvic-hip complex during running can be measured with sufficient repeatability to justify the implementation of the method as an evaluation tool in future investigations. However, caution must be exerted when interpreting the absolute magnitudes of trunk flexion-extension, lumbar spine flexion-extension, pelvic anterior-posterior tilt, hip axial rotation and thigh axial rotation, given the poor level of repeatability found for these measures. PMID- 11869908 TI - Medial patellar taping changes the peak plantar force location and timing of female basketball players. AB - Subtle changes in patellofemoral joint biomechanics may influence lower extremity function. The McConnell method of patellar taping has become an increasingly popular method of managing patients with various patellofemoral joint disorders. The purpose of this order effect controlled study was to assess medial patellar taping (McConnell Method) for changes in peak plantar force location and timing while running and dribbling a basketball prior to the performance of a basketball lay-up. Sixteen non-impaired, right hand dominant members of a female junior varsity basketball team (age=14.6 +/- 2 years) participated in this study. Subject bodyweight and height were 607.8 +/- 99 N and 1.67 +/- 0.10 m, respectively. All data were collected from the preferred stance limb. A series of two way analysis of variance (ANOVA) (condition, trial) were used to determine statistical significance (P < or = 0.05). During medial patellar taping, subjects displayed a more forefoot directed peak plantar force location 89.9 +/- 18 versus 81.3 +/- 21 mm and delayed peak plantar force onset 141 +/ - 23 versus 130 +/- 29 ms following initial ground contact. Medial-lateral peak plantar force location and peak plantar force magnitude did not differ between conditions, however, medial-lateral peak plantar force location displayed significant trial sequence influences with the latter trials displaying more lateral center of plantar force locations (30.4 +/- 2 vs. 32.5 +/- 3 mm). The results of this study suggest that medial patellar taping influences distal lower extremity function by shifting peak plantar force location toward the forefoot, and delaying its onset. These changes with consideration for known synergistic ankle plantar flexor-subtalar joint invertor and knee extensor function during weight bearing suggest the presence of increased muscular stiffness acting through a more rigid foot to improve the impact force attenuating capability of the lower extremity. PMID- 11869909 TI - Ground reaction forces and frictional demands during stair descent: effects of age and illumination. AB - Stair descent is an inherently risky and demanding task that older adults often encounter in everyday life. It is believed that slip between the foot or shoe sole and the stair surface may play a role in stair related falls, however, there are no reports on slip resistance requirements for stair descent. The aim of this study was to determine the required coefficient of friction (RCOF) necessary for safe stair descent in 12 young and 12 older adults, under varied illuminance conditions. The RCOF during stair descent was found to be comparable in magnitude and time to that for overground walking, and thus, with adequate footwear and dry stair surfaces, friction does not appear to be a major determinant of stair safety. Illuminance level had little effect on the dependent variables quantified in this study. However, the older participants demonstrated safer strategies than the young during stair descent, as reflected by differences in the ground reaction forces and lower RCOF. PMID- 11869910 TI - Changes in gait when anticipating slippery floors. AB - Falls precipitated by slipping are listed among the leading causes of injuries. The biomechanical analysis of such events is a necessary component of the slips/falls prevention research. One of the challenges of biomechanical studies is reproducing the unexpected nature of real-life slipping accidents. Thus, the goal of this study was to quantify changes in gait biomechanics when subjects anticipate slippery environments. Foot ground reaction forces and body dynamics of 16 subjects were recorded during level walking and descending ramps of varying frictional properties and inclination. Gait biomechanics were compared among three types of dry trials: (1) baseline (subjects knew the floor was dry); (2) anticipation (subjects were uncertain of the contaminant condition, dry, water, soap or oil); and (3) recovery trials recorded after a contaminated trial (subjects again knew the floor was dry). Subjects were asked to walk as naturally as possible throughout testing. Anticipation trials produced peak required coefficient of friction (RCOF(peak)) values that were on average 16-33% significantly lower than those collected during baseline trials, thus reducing slip potential. During recovery trials, RCOF(peak) values did not return to baseline characteristics (5-12% lower). Postural and temporal gait adaptations, which affected ground reaction forces, were used to achieve RCOF(peak) reductions. Statistically significant gait adaptations included reductions in stance duration (SD) and loading speed on the supporting foot, shorter normalized stride length (NSL), reduced foot-ramp angle and slower angular foot velocity at heel contact. As a result of these adaptations, anticipation of slippery surfaces led to significant changes in lower extremity joint moments, a reflection of overall muscle reactions. Thus, this study suggests that significant gait changes are made when there is a potential risk of slipping even though subjects were asked to walk as naturally as possible. Insights are also gained into the adaptations that are used to reduce the potential of slips/falls. PMID- 11869911 TI - Periods of extreme ankle displacement during one-legged standing. AB - The goal of this study was to describe the movement patterns of the ankle joint whilst standing on one leg. Ten healthy adult females (age 24 +/- 5.3 years) performed a one-legged standing task with eyes closed. Force platform recordings and video analyses were used to describe the kinematic and kinetic characteristics of the ankle joint during this task. A rocking movement of the foot (heel inversion-eversion) was documented by examining instances of extreme ankle displacement. Extreme ankle displacement was defined as any instant when the ankle position was more than +/- 2 SD away from the mean ankle joint position. Extreme values of lateral and medial ankle joint displacement were 14.8 and 9.2 mm, correspondingly. These instances of extreme foot inversion-eversion were characterized by large medial-lateral displacement of the gravity line (GLP) and center of pressure (COP) and large horizontal forces. Comparing instances of extreme ankle joint displacement to periods of non-extreme ankle displacement, the ankle joint moment remained fairly constant, averaging 8.4 +/- 4 and 6.9 +/- 3.5 Nm, respectively. The moment about the 'body-minus-foot' center of mass generated by the ankle joint reaction force, however, was on average over four times larger during instances of extreme ankle displacement (3.4 +/- 2.8 Nm), than during periods of non-extreme ankle displacement (0.8 +/- 0.4 Nm). In utmost situations, the moment due to the joint reaction force was up to 73% of the ankle joint moment. These results suggest that at least two different techniques are used to maintain balance during one-legged standing. The first technique, termed the ankle torque technique, involves a large restorative moment at a stationary ankle joint for balance maintenance. The other technique, the shear force technique, involves a large horizontal force at a moving ankle joint for balance maintenance. During non-extreme periods, balance was maintained primarily through the ankle torque technique. During extreme instances, a combination of both techniques was observed. PMID- 11869912 TI - Identification of individual walking patterns using time discrete and time continuous data sets. AB - Scientific studies typically treat data by studying effects of groups. Clinical therapy typically treats patients on a subject specific basis. Consequently, scientific and clinical attempts to help patients are often not coordinated. The purposes of this study were (a) to identify subject and group specific locomotion characteristics quantitatively, using time discrete and time continuous data and (b) to assess the advantages and disadvantages of the two approaches. Kinematic and kinetic gait pattern of 13 female subjects walking in dress shoes with different heel heights (14, 37, 54 and 85 mm) were analysed. The results of this study showed that subject specific gait characteristics could be better identified with the time continuous than with the time discrete approach. Thus, the time continuous approach using artificial networks is an effective tool for identifying subject and group specific locomotion characteristics. PMID- 11869913 TI - Judo, better than dance, develops sensorimotor adaptabilities involved in balance control. AB - OBJECTIVES: Training allows sportsmen to acquire new balance control abilities, possibly differing according to the discipline practised. We compared, by means of static and dynamic posturographic tests, the postural skills of high-level judoists, professional dancers and controls, in order to determine whether these sports improved postural control. RESULTS: With eyes open, judoists and dancers performed better than controls, indicating a positive effect of training on sensorimotor adaptabilities. Yet, with eyes closed, only judoists retained a significantly better stance. CONCLUSIONS: These data indicate that the practice of a high-skill activity involving proprioceptive afferences especially improves both performance and balance control. PMID- 11869914 TI - Leg length discrepancy. AB - The role of leg length discrepancy (LLD) both as a biomechanical impediment and a predisposing factor for associated musculoskeletal disorders has been a source of controversy for some time. LLD has been implicated in affecting gait and running mechanics and economy, standing posture, postural sway, as well as increased incidence of scoliosis, low back pain, osteoarthritis of the hip and spine, aseptic loosening of hip prosthesis, and lower extremity stress fractures. Authors disagree on the extent (if any) to which LLD causes these problems, and what magnitude of LLD is necessary to generate these problems. This paper represents an overview of the classification and etiology of LLD, the controversy of several measurement and treatment protocols, and a consolidation of research addressing the role of LLD on standing posture, standing balance, gait, running, and various pathological conditions. Finally, this paper will attempt to generalize findings regarding indications of treatment for specific populations. PMID- 11869915 TI - Dealing with rejection. PMID- 11869916 TI - Tanner's pubic hair staging is not applicable to all adolescents. PMID- 11869917 TI - Female adolescents and older male sex partners: HIV associated risk. PMID- 11869918 TI - Psychometric properties of CAP. PMID- 11869920 TI - Prevalence, attitudes, and behaviors related to the use of nonsteroidal anti inflammatory drugs (NSAIDs) in student athletes. AB - PURPOSE: To describe NSAID use among football players and examine differences in attitudes and behaviors associated with reports of daily use. Data are from 681 self-administered questionnaires. Independent variables were race (white), insurance status (private), attitudes (whether NSAIDs helped performance), and behaviors (decided independently vs. asked an adult when to take an NSAID, what type, or dose; use of NSAIDs to treat pain vs. block pain). Among NSAID users, independent variables were entered into a logistic regression to determine their effects on the probability of daily use. Athletes were male, 90% white, mean age 15.8 years, and 88% had private insurance. Four-hundred-fifty-two (75%) used NSAIDs in the past 3 months, and 90 (15%) were daily users with no significant difference in age, race, or insurance between users and nonusers. Those who perceived better performance owing to NSAIDs (adjusted odds ratio [AOR] = 2.4), who decided on their own when to take NSAIDs (AOR = 2.2), and who used NSAIDs prophylactically were more likely to use daily NSAIDs (AOR = 2.5) after controlling for other variables in the model. NSAID use in high school football players is highly prevalent. Although package inserts caution against chronic use, nearly one of seven players uses NSAIDs daily. Daily users were more likely to decide independently and to use NSAID prophylactically, raising issues about the need for adult supervision and education of athletes regarding side effects and appropriate use. PMID- 11869921 TI - Telephone vs. face-to-face notification of HIV results in high-risk youth. AB - PURPOSE: To increase the number of high-risk and homeless youth who receive human immunodeficiency virus (HIV) test results and posttest counseling. METHODS: Oral HIV testing and counseling were offered to high-risk and homeless youth at sites at which youth congregate throughout Portland, Oregon. Subjects were randomized to receive test results and posttest counseling either in a face-to-face manner or with the option of telephone notification. Self-reported demographic and risk behavior information was collected prior to HIV testing. The differences in the proportion of youth who received their test results were analyzed according to the notification method and demographic characteristics using SPSS. RESULTS: Among the 351 youth who were tested, 48% followed up to receive test results and posttest counseling. Adolescents most likely to receive their results were female, older (19-24 years), and white and those who reported high-risk behaviors. Those given the option of telephone notification were significantly more likely to receive their results than those required to have face-to-face notification (odds ratio = 2.301, 95% confidence interval of 1.499, 3.534). This was true regardless of age, race, history of previous HIV testing, or presence of high-risk behaviors. Two youths tested positive for HIV corroborating previous reports of low HIV prevalence in this population. Both were assigned to the face to-face notification group and, therefore, no HIV positive results were given by telephone. CONCLUSIONS: The option of telephone notification significantly increased the proportion of youth who received posttest counseling and results following community-based testing. PMID- 11869922 TI - From adolescence through young adulthood: psychosocial adjustment associated with long-term survival of HIV. AB - PURPOSE: To examine the psychosocial factors associated with long-term survival of pediatric human immunodeficiency virus (HIV) infection. METHODS: Children infected with HIV enrolled in clinical trials at the National Cancer Institute and their caregivers were interviewed and completed self-report measures 3 times, approximately 12 months apart, using the Child Behavior Checklist, Social Support Scale for Children, Self-Perception Profile for Children and Adolescents, and a structured interview designed by the investigators. Historical data were also extracted from patient medical charts. Average age of participants was 11.8 years at time 1 and 14 years at time 2; 56.3% of the original sample were male, racial composition was 72.2% white, 13.9% African-American, 6.9% Hispanic, and 6.9% "other"; 38.9% of participants contracted HIV perinatally, 34.7% through a hemophilia-related transfusion, and 26.4% through another type of transfusion. RESULTS: Pearson product-moment correlations revealed that disclosure was found to be positively related to social support, self-competence, and decreased problem behavior, except in the case of public disclosure, in which an independent-sample Student's t-test revealed that it was negatively associated with global self-competence. Social support was significantly negatively correlated with problem behavior. Chi-square analyses of the 5-year follow-up data indicated that participants aged 18 years and older were less likely to complete their academic education than their healthy peers (national norms). Adolescents who lost a parent were more likely to have suffered from depression during their lifetime. CONCLUSIONS: Social support and open communication about the diagnosis are essential, particularly at an age at which decisions about relationships, sexual activity, drug use, and plans for the future are the focus of adolescent development and individuation. With advances in medical treatment, HIV-infected children are more likely to survive into adolescence and beyond. Accordingly, their psychosocial needs are changing to more closely resemble the needs of the chronically ill individual, rather than the terminally ill. Families of HIV-infected children should seriously consider preparation for independent living. PMID- 11869924 TI - The relationship between smoking, drinking, and adolescents' self-perceived health and frequency of hospitalization: analyses from the 1997 National Household Survey on Drug Abuse. AB - PURPOSE: To investigate the more immediate health effects of smoking and drinking among adolescents. METHODS: Secondary analyses of data from the 1997 National Household Survey on Drug Abuse were conducted to explore the relationship between subjective and objective health outcomes and the use of alcohol and tobacco among adolescents. RESULTS: The findings suggest that adolescents who smoke or drink actually report poorer health during adolescence than those who do not. In fact, adolescents who are frequent or heavy alcohol and tobacco users report poorer subjective overall health and a greater number of overnight hospital stays during the previous year than less frequent or intense users and than nonusers. In addition, the data revealed a number of significant gender and age group differences in reports of health outcome as well. CONCLUSIONS: Demonstrating to youngsters the more immediate, negative health consequences of alcohol and tobacco use might strengthen prevention messages and ultimately decrease the use of these substances among adolescents. PMID- 11869923 TI - Increased prevalence of herpes simplex virus type 2 among adolescent women with alcohol use disorders. AB - PURPOSE: To determine the relationship between alcohol use disorders (AUDs) and herpes simplex virus type 2 (HSV-2), hepatitis B virus (HBV), and human immunodeficiency virus (HIV) infections among a sample of sexually active adolescents. METHODS: Subjects were 240 sexually active male and female adolescents aged 14 to 21 years (mean 17.5 years) recruited from clinical and community settings in western Pennsylvania between 1991 and 1995; 55% had a lifetime history of AUDs (63 females, 69 males) and 45% did not have a lifetime history of AUDs (57 females, 51 males). Participants provided information about demographic factors and sexual behaviors as well as a serum sample that was assayed for antibodies to HSV-2, HBV, and HIV infections. Multivariate logistic regression was used to determine the independent relationship of AUDs to HSV-2 infections among females. RESULTS: The seroprevalence of HSV-2 infections was 15% among females and 0% among males; the overall prevalence of HBV (1.2%) and HIV (0.4%) infections was very low. Among adolescent females, the seroprevalence of HSV-2 infections was significantly higher among those with an AUD (19%), compared with those without an AUD (10.5%) (adjusted odds ratio 8.1, 95% confidence interval 1.5-44.8, p =.017). CONCLUSIONS: Adolescent women with an AUD appear to be at substantially increased risk of HSV-2 infection. These results highlight the need to address sexually transmitted diseases among adolescents with alcohol problems. PMID- 11869925 TI - Family processes for children in early elementary school as predictors of smoking initiation. AB - OBJECTIVES: To assess the relationship between family processes measured when children are in early elementary school and initiation of cigarette smoking in early adolescence. METHODS: The analysis sample of 810 children was drawn from a longitudinal study of students from a suburban school district in the Pacific Northwest. Predictor variables were assessed when children were in second or third grade, and smoking initiation was measured when the children were in sixth or seventh grade. Measures of family processes were entered separately into logistic regression models that included controls for household structure and income, parent smoking, and peer and child characteristics. RESULTS: Measures of child attachment to parent and parent involvement with the child's school were significantly and negatively associated with smoking initiation. Among control variables, parent smoking, child grade level, and child antisocial behavior and depression were the strongest predictors of smoking initiation. CONCLUSIONS: The results point to the importance of family bonding and parent supportiveness as protective factors and parent smoking and early childhood antisocial behavior and depression as risk factors for smoking initiation in pre- or early adolescence. PMID- 11869927 TI - The patient health questionnaire for adolescents: validation of an instrument for the assessment of mental disorders among adolescent primary care patients. AB - PURPOSE: To investigate the validity of the Patient Health Questionnaire for Adolescents (PHQ-A), a self-administered instrument that assesses anxiety, eating, mood, and substance use disorders among adolescent primary care patients. METHODS: A total of 403 adolescents from California, New Jersey, New York, and Ohio completed the PHQ-A and the Medical Outcomes Study Short-Form General Health Survey (SF-20) during or shortly after a visit to a primary care clinic or a school nurse's office. A few days later, clinical psychologists who were blind to the results of the PHQ-A administered a semi-structured clinical interview to assess the same psychiatric disorders and to conduct a global assessment of functioning (GAF) among 403 patients. Diagnostic agreement coefficients were computed and analyses of covariance were conducted. RESULTS: Findings support the diagnostic validity of the PHQ-A. The PHQ-A and the clinical interview produced similar estimates of the prevalence rates of anxiety, eating, mood, and substance use disorders. The PHQ-A demonstrated satisfactory sensitivity, specificity, diagnostic agreement, and overall diagnostic accuracy, compared with the clinical interview. Adolescents with PHQ-A diagnoses experienced significantly poorer mental and overall functioning, more physical pain, and poorer overall health compared with those without psychiatric disorders. These differences remained significant after patients' age, gender, ethnicity, and site were controlled statistically. CONCLUSION: The PHQ-A may be used to assist primary care practitioners in identifying psychiatric disorders among their adolescent patients. The PHQ-A is the first such tool to be tested for use in adolescents and offers an acceptable and efficient tool for early detection and recognition of mental disorders in this high-risk group. PMID- 11869926 TI - Delivering quality care: adolescents' discussion of health risks with their providers. AB - PURPOSE: To compare adolescents' report of topics they wanted to discuss with their providers with what they actually discussed, and whether they talked to their providers about their self-reported health risks. METHODS: We analyzed the 1997 Commonwealth Fund Survey of the Health of Adolescent Girls, a nationally representative sample of 6728 in-school adolescent boys and girls in 5th through 12th grade. Respondents reported on health risks, whether they believed their provider should discuss certain topics, and whether their provider did talk about health-related topics. Logistic regression was used to compare proportions and assess the associations among variables. RESULTS: Adolescents most frequently discussed healthy dietary habits (49%), weight (43%), and exercise (41%) with their clinicians but most frequently wanted to discuss drugs (65%), smoking (59%), and healthy dietary habits (57%). Overall, 70.9% of the sample reported at least one of eight potential health risks, but 63% of these adolescents had not spoken to their doctor about any of these risks. Using logistic regression models, having a female provider (odds ratio [OR] 1.41), obtaining health care information from a doctor (OR 1.72) and from the Internet (OR 1.50), speaking privately with their provider (OR 1.45), and reporting more total risk factors (OR 1.59) were each associated with having discussed any risks. CONCLUSIONS: Adolescents want and need to discuss health care issues with their providers but often do not. Providing quality preventive care to adolescents will require increased physician screening and counseling about these issues. PMID- 11869928 TI - Stature and pubertal stage assessment in American boys: the 1988-1994 Third National Health and Nutrition Examination Survey. AB - PURPOSE: To describe current stature and pubertal development in North American boys, and to compare these measures with measures observed approximately 30 years ago. METHODS: We analyzed data (i.e., height, weight, and Tanner Stage) from the Third National Health and Nutrition Examination Survey (NHANES III), conducted between 1988-1994, and compared it to the National Health Examination Survey, Cycles II and III (HES II/III), conducted from 1963-1965 and 1966-1970. The surveys included physical examination and questionnaire components, employed cross-sectional designs, and are nationally representative. We used logistic regression to calculate median age at onset of pubertal stages. RESULTS: NHANES III included 2481 boys aged 8 to 18 years. HES II comprised 3010 boys aged 8-11 years and HES III comprised 3514 boys aged 12-17 years. The mean heights of the oldest boys in both surveys did not differ significantly; however, at younger ages, boys in the more recent survey were taller (average height difference among those aged 8-14 years was 2.0 cm). Boys in NHANES III were also heavier and had higher body mass index than those in HES II/III. The median estimated ages of onset of pubertal stages in NHANES III were 9.9, 12.2, 13.6, and 15.8 years for genital stages 2-5, respectively, and 11.9, 12.6, 13.6, and 15.7 years for pubic hair stages 2-5, respectively. For some stages, the median estimated age of onset of puberty was earlier among boys in NHANES III than among those in HES III. CONCLUSIONS: Differences in mean height at young ages, but not at older ages, suggest that the rate of growth among boys in NHANES III was faster than that of boys in the earlier surveys. This finding, coupled with the finding of earlier ages of onset of some pubertal stages, suggests that boys of this generation may be maturing more rapidly than did boys in the past. PMID- 11869929 TI - Lead exposure and urinary N-acetyl beta D glucosaminidase activity in adolescent workers in auto repair workshops. AB - PURPOSE: To investigate levels of lead (Pb) exposure and renal tubular damage among adolescent workers in auto repair workshops in Turkey. METHODS: The study was conducted on 39 adolescent workers (mean age: 16.18 +/- 3.19 years) in auto repair workshops (8 autoelectrician, 10 motor repairman, 8 auto painter, 5 turner, 8 bonnet straighter). Thirteen adult employees of battery production in the workshops (mean age: 32.08 +/- 10.94 years) and 29 healthy rural adolescent (mean age: 14.78 +/- 2.68 years) constituted the control groups. The level of blood Pb was investigated by an atomic absorption spectrophotometer and urinary N acetyl beta- D glucosaminidase (NAG) activity was measured by a colorimetric method. Mann-Whitney U test was performed to examine group differences. RESULTS: All subjects and controls had normal blood urea, creatinine, uric acid, sodium, potassium levels, normal routine urine examination and tubular phosphorus reabsorption. Blood Pb levels in auto repair workers (8.13 +/- 7.41 mug/dL) were significantly higher than the rural control group (3.49 +/- 1.39 mug/dL) but lower than the battery workers (25.27 +/- 9.82 mug/dL). Urinary NAG (U/gr creatinine) (4.71 +/- 2.11) was lower than the battery workers (7.39 +/- 4.37), however significantly higher than the normal control group (3.07 +/- 1.20). In addition, auto painters had higher levels of Pb exposure and urinary NAG activity than the other workers (p <.05). CONCLUSION: Chronic low dose Pb exposure was found to cause renal tubular injury in children workers of auto repair workshops. PMID- 11869930 TI - A strategy to deter blood doping in sport. PMID- 11869931 TI - Further concerns about the medical risks of blood doping. PMID- 11869932 TI - Thalidomide in multiple myeloma: state of art. PMID- 11869933 TI - Genomic instability at the human CD5 gene promoter. AB - BACKGROUND AND OBJECTIVES: The gene coding for the human CD5 lymphocyte surface receptor maps to the 11q12.2 region, which is in the vicinity of a region commonly affected by multiple somatic mutations in human cancers. The 5'-flanking region of the human CD5 gene includes an evolutionarily conserved (TC)n(CA)n microsatellite (MS) of potential utility as a marker for genome instability. The aim of the present study was to investigate the value of the CD5 MS as a marker for instability in different tumor types, particularly in B-cell leukemia and lymphoma. DESIGN AND METHODS: The CD5 MS and a panel of 10 MS markers were analyzed by using a polymerase chain reaction (PCR)-based method and polyacrylamide sequencing gels. This was done in several hematopoietic and non hematopoietic cell lines, as well as in 28 cases of B-cell chronic lymphocytic leukemia (B-CLL), 19 mantle cell lymphomas (MCL) and 45 head and neck carcinomas (HNC). RESULTS: The frequency of CD5 MS abnormalities found among HNC was similar to that reported for other well known MS markers at loci near known and suspected cancer genes. However, instability at the CD5 MS was the most frequent MS abnormality among B-CLL and MCL. INTERPRETATION AND CONCLUSIONS: The inclusion of MS markers at chromosome 11q may be especially informative for genome instability analyses of certain B-cell leukemias and lymphomas. PMID- 11869934 TI - H63D mutation in the HFE gene increases iron overload in beta-thalassemia carriers. AB - BACKGROUND AND OBJECTIVES: Hereditary hemochromatosis (HH) is an autosomal recessive disorder of iron metabolism. The HFE gene implicated in this disorder has been identified on chromosome 6 (6p21.3). The most prevalent mutation in HH patients changes the 282 cysteine residue to tyrosine (C282Y). The role of a second mutation which changes the 63 histidine to aspartic acid (H63D) in iron overload has been controversial. The aim of this study was to evaluate the effect of the H63D mutation on the ferritin levels of beta-thalassemia carriers. DESIGN AND METHODS: beta-thalassemia carriers have a tendency to increase iron absorption because of mild anemia and slightly increased erythropoiesis. Differences in ferritin levels between homozygotes for H63D and wild type may indicate a modulator effect of the HFE mutation on iron absorption. We studied 152 healthy males, heterozygous for beta-thalassemia. Serum ferritin was measured by chemiluminescence. H63D genotypes were determined by digestion of polymerase chain reaction (PCR) products with MboI restriction enzyme. RESULTS: Forty-five subjects were H63D heterozygotes and four subjects were H63D homozygotes. Ferritin levels were (mean +/- SD): 250 +/- 138 microg/L in homozygotes for the wild type H/H; 295 +/- 186 microg/L in H/D heterozygotes; and 389 +/- 75 microg/L in homozygotes for the mutation D/D. The difference in ferritin values between H/H and D/D is statistically significant (p=0.022). INTERPRETATION AND CONCLUSIONS: beta-thalassemia carriers who are homozygotes for the H63D mutation have higher ferritin levels than beta-thalassemia carriers with the H/H genotype, suggesting that the H63D mutation may have a modulating effect on iron absorption. PMID- 11869935 TI - DNAase I hypersensitive site 3' to the beta-globin gene cluster contains a TAA insertion specific for beta(S)-Benin haplotype. AB - BACKGROUND AND OBJECTIVES: Analysis of DNA polymorphic sites is a powerful tool for detection of gene flow in human evolutionary studies and to trace genetic background associated with abnormal genes. The beta-globin locus contains more than 20 single-base restriction fragment length polymorphism (RFLP) sites spanning over 80 kb on chromosome 11. Far downstream of the expressed genes, there is a hypersensitive site (HS). The function of the 3'-HS remains unknown. As an approach to the understanding of the 3'-HS region in sickle cell anemia we searched for sequence polymorphism in the AT-rich region, using a non-radioactive polymerase chain reaction (PCR)-single strand conformational polymorphism (SSCP) technique. DESIGN AND METHODS: A 460 bp fragment located at the 3' of the b globin gene was amplified from patients (with sickle cell anemia and HbSC disease), and from AS individuals. Standard RFLP-haplotyping was performed and compared with the PCR-SSCP screening strategy. RESULTS: Two distinct band patterns were revealed by SSCP testing, each one in strict linkage disequilibrium with either Benin or Bantu haplotypes. Direct sequencing of the amplified segment revealed a TAA insertion in the AT-rich region, in all 121 beta(S) Benin chromosomes tested, but not in other beta(S) haplotypes from the total of 380 beta(S) chromosomes typed. INTERPRETATION AND CONCLUSIONS: SSCP analysis could easily distinguish sequence variations in the 3'AT-rich region of the beta-globin cluster, and a TAA insertion in this region seems to be specific for the Benin beta(S) chromosome. PMID- 11869936 TI - Prognostic significance of CD56 antigen expression in acute myeloid leukemia. AB - BACKGROUND AND OBJECTIVES: CD56 antigen expression has been reported in several hematologic malignancies. In acute myeloid leukemia (AML)M2 with t(8;21) and acute promyelocytic leukemia (APL) it has been found to be consistently associated with an unfavorable prognosis, whereas in other AML subtypes its role remains uncertain. We investigated CD56 expression in a cohort of AML patients in order to assess its frequency and prognostic relevance. DESIGN AND METHODS: Immunophenotypic analysis including that of CD56 antigen was available for 171 consecutive AML patients (139 with AML and 32 with APL), enrolled between December 1995 and December 1999 at a single institution. A sample of fresh bone marrow cells taken at diagnosis was recorded as positive when at least 20% of the cells double-stained with specific monoclonal antibodies against CD56 and CD33 antigens. RESULTS: CD56 positivity was demonstrated in 37 cases (21.6%). Its frequency was lower in M4 (6%) and higher in M5 (37%). The median percentage for CD56+ blasts was 56% (range 21-99%). CD56 positivity did not correlate with age, sex, blast count, favorable or unfavorable cytogenetics at diagnosis, nor did it influence the outcome in terms of complete remission (CR) duration (606 vs. 417 days, p=n.s.) or overall survival (OS) (210 vs. 277 days, p= n.s.). In the APL subgroup a significant difference in relapse rate was found at 3 years (71.4% in the CD56 positive group vs. 12% in the CD56 negative group, p=0.005). INTERPRETATION AND CONCLUSIONS: Our data confirm that CD56 positivity in APL patients at diagnosis is associated with a worse prognosis, suggesting that close molecular monitoring is necessary in CD56 positive APL patients. In contrast, the prognostic role of CD56 remains uncertain in the other AML subtypes. PMID- 11869938 TI - Acquired von Willebrand factor abnormalities in myeloproliferative disorders and other hematologic diseases: a retrospective analysis by a single institution. AB - BACKGROUND AND OBJECTIVES: Acquired von Willebrand syndrome (AVWS) is a rare acquired disorder. In most cases it is associated with lymphoproliferative disorders and monoclonal gammopathies, while less frequently myeloproliferative disorders (MPD) are involved. Although bleeding is the most important symptom, thrombotic complications have also been observed in cases associated with MPD. Our aim was to review the clinical and laboratory findings in AVWS patients from a single institution. DESIGN AND METHODS: The records of 99 patients with AVWS were reviewed to identify the underlying diseases, the symptoms and the laboratory parameters. RESULTS: In 75% of cases the AVWS was associated with MPD. The most frequent pattern was type 2 (67.7%). Abnormalities of bleeding time, factor VIII levels or platelet retention to glass beads were observed in 83.8% of cases. Bleeding was present in 38.4% of patients, more frequently in the not-MPD associated (58.3%) vs. MPD-associated cases (32%) (p=0.022), with a significant predominance in females, irrespective of the underlying disease (p=0.0007). In 32% of patients with MPD, thrombotic manifestations, mostly microvascular and arterial episodes, were observed. INTERPRETATION AND CONCLUSIONS: AVWS in MPD seems to be mainly a laboratory diagnosis, without clinical symptoms in most cases, although bleeding as well as ischemic events can be present. In contrast, AVWS in not-MPD-associated cases is most frequently associated with severe bleeding symptoms. Performing appropriate laboratory tests may be useful for screening for AVWS. PMID- 11869939 TI - A chromogenic substrate method for detecting and titrating anti-factor VIII antibodies in the presence of lupus anticoagulant. AB - BACKGROUND AND OBJECTIVES: The development of neutralizing anti-factor VIII antibodies (a-fVIII) is a major clinical complication. Lupus anticoagulant (LA) might affect detection of a-fVIII, since both inhibitors may act on the same coagulation pathway. Our aim was to accomplish unequivocal detection and titration of a-fVIII even in the presence of LA. DESIGN AND METHODS: We evaluated a-fVIII activity by a chromogenic substrate (CS) method in samples with a-fVIII (n=6), LA (n=12) and presumably both LA+a-fVIII (n=5). The inhibition index before (Ii) and after incubation at 37 C (Ii(37)) was estimated. We also performed factor VIII assays (one-stage and CS) and titration methods (Bethesda and CS) in parallel. RESULTS: Inhibition in the a-fVIII group (Ii=5-3200) was potentiated by incubation (Ii(37)=27-5200) as it was in LA+a-fVIII (Ii=9-21; Ii(37)=50-903). LA samples showed no or meaningless inhibitory effect (Ii=0-7; Ii(37)=0-4) or a-fVIII activity (0.00-0.06 CSU/ml) by the CS method; on the contrary, very low to moderate (0.52-7.00 BU/ml) a-fVIII activity was recorded by the Bethesda method. The two titration methods did not correlate (p>0.100) in the presence of LA, or LA+a-fVIII. Differences between factor VIII:C and factor VIIIcs were significant only in LA samples (p=0.005); however, patients with residual factor VIII activity from the LA+a-fVIII group also showed higher factor VIIIcs values than factor VIII:C ones. INTERPRETATION AND CONCLUSIONS: Results indicate the possibility of detecting and titrating a-fVIII without interference of LA by the CS method. This marks a difference with respect to the Bethesda method, in which a measurable effect can be expected in the presence of a strong LA. PMID- 11869937 TI - Phase I trial with escalating doses of idarubicin and multidrug resistance reversal by short-course cyclosporin A, sequential high-dose cytosine arabinoside, and granulocyte colony-stimulating factor for adult patients with refractory acute leukemia. AB - BACKGROUND AND OBJECTIVES: Patients with refractory acute myeloid or lymphoid leukemia (AML, ALL) were treated with a high-dose regimen comprising idarubicin (IDR) plus short-course cyclosporin A (CsA) as multidrug resistance type-1 (MDR1) blocking agent. The principal aim was to define the maximum tolerated dose (MTD) of IDR, which is reported to be a less MDR1-sensitive anthracycline. The short CsA infusion was patterned after the results of a previous in vitro study. DESIGN AND METHODS: This was a phase I trial, in which eligible patients received high dose cytarabine (HDAC) 3 g/m(2)/bd on days 1, 2 and 8, 9, and IDR 12.5-20 mg/m(2)/d on days 3 and 10, with increments of 2.5 mg/m(2)/d from the baseline per treatment group. Intravenous CsA infusion started 4 hours before IDR and lasted 12 hours. Recombinant granulocyte colony-stimulating factor (G-CSF) was added from day 11. IDR MTD was evaluated through analysis of regimen-related toxicity (RRT). RESULTS: Eighteen patients were treated (16 AML, 2 ALL; MDR1+: 8/8 studied). Overall response rate was 61%. Toxicity was severe but manageable up to an IDR dose of 17.5 mg/m(2)/d, while grade 4 RRT developed with IDR 20 mg/m(2)/d. High-grade toxicity, not strictly regimen-related, was sometimes observed at lower IDR concentrations in patients with unresolved complications from prior extensive treatments. In keeping, the complete response (CR) rate was 92% (11/12) for patients with an ECOG performance score <2 compared to 0% (0/6) in the others (p=0.000). Apart from that, induction of markedly hypocellular, leukemia-free bone marrow on day 11 was associated with achievement of CR (13 evaluable: CR 8/10 vs 0/3, p=0.035). INTERPRETATION AND CONCLUSIONS: IDR at 17.5 mg/m(2)/d (x2) can be associated with short-course CsA and HDAC for the management of refractory acute leukemias. While this regimen could deserve testing in a larger phase II trial, to document activity in MDR1+ disease, it remains important to select the most suitable patients in order to avoid the occurrence of life-threatening cumulative toxicity. PMID- 11869940 TI - The prothrombin 20210A allele influences clinical manifestations of hemophilia A in patients with intron 22 inversion and without inhibitors. AB - BACKGROUND AND OBJECTIVES: The modulation of disease severity in hemophilia A (HA) patients may be related to the co-inheritance of mutations in genes with a known thrombotic effect such as factor V Leiden (FVL) and prothrombin. In the Spanish population, the prothrombin 20210A (PT20210A) allele is the most prevalent genetic risk factor for venous thromboembolism. DESIGN AND METHODS: We investigated the presence of both mutations in a cohort of 265 hemophiliac patients divided into two groups: I) 140 unrelated patients with moderate and mild HA and II) 125 unrelated patients with severe HA (83 carrying an inversion of intron 22). RESULTS: In group I, 4 patients had the FVL (2.8% vs. 2.98% controls) and 5 had the PT20210A (3.6% vs. 6.46% controls). In group II, two patients with inversion had the FVL (1.6%) and PT20210A was found in 10 patients (8%), five of them with inversion of intron 22 without inhibitors. One of these patients had the FVL and PT20210A mutations concomitantly. In the subgroup of patients with inversion who were carriers of the PT20210A, three parameters i.e. spontaneous bleeding (p=0.008), factor VIII utilization (p=0.016) and number of hemophilic arthropathies (p<0.0005) were significantly lower than in a subgroup of 11 age-matched non-PT20210A severe HA patients with inversion and without inhibitors. INTERPRETATION AND CONCLUSIONS: These results indicate that the inheritance of PT20210A could be a protective factor that mitigates the clinical severity of HA. PMID- 11869941 TI - Selecting patients for home treatment of deep vein thrombosis: the problem of cancer. AB - BACKGROUND AND OBJECTIVES: Patients with deep vein thrombosis are selected for home treatment on the basis of their clinical and social condition. Cancer is frequently associated with venous thromboembolism and is often considered an exclusion criterion for outpatient treatment. We investigated the impact of cancer on the outpatient management of venous thrombosis. DESIGN AND METHODS: We performed a prospective, cohort study on consecutive patients with objectively documented deep vein thrombosis. All were assessed for home treatment. Hospital admission was recommended in the presence of common exclusion criteria. All patients were treated with low molecular weight heparin and warfarin. Information on previous, active, or suspected cancer was collected. Recurrent thrombosis, bleeding and mortality were documented at a 3-month follow-up. RESULTS: One hundred patients were included; 72 were entirely treated at home (mean age: 61.2 years). There were 22 patients with known cancer: 12 (55%) were managed as outpatients (16.5% of the outpatient population) and 10 were hospitalized (36% of the inpatient population), 6 because of a poor clinical condition, 4 because further investigation of their malignancy was required. The presence of cancer and the likelihood of poor compliance were the most frequent reasons cited for in hospital treatment. Overall, event rates at 3 months were comparable to those reported in previous studies in the outpatient population and slightly higher in the inpatient population (recurrent thrombosis 1.5% and 7%; bleeding 5.5% and 10.7%; mortality 4% and 18%, respectively). INTERPRETATION AND CONCLUSIONS: Cancer was the most common reason cited for in-hospital treatment. Nevertheless, more than half of the patients with known cancer were safely and effectively treated at home. PMID- 11869942 TI - Predicting factors for admission to an intensive care unit and clinical outcome in pediatric patients receiving hematopoietic stem cell transplantation. AB - BACKGROUND AND OBJECTIVES: In children, hematopoietic stem cell transplantation (HSCT) implies life-threatening complications and some patients need admission to a pediatric intensive care unit (PICU). Few studies have been reported analyzing this issue in a pediatric population and most focused on risk factors predicting survival following PICU admission. DESIGN AND METHODS: We examined data of 240 pediatric patients who received HSCT (100 allogeneic and 140 autologous) in order to ascertain the incidence of life-threatening complications requiring PICU admission, the contributing risk factors and the patients' long-term survival. RESULTS: Forty-two (17.5%) (25 males and 17 females) of the transplanted children were admitted to the PICU. Twenty-nine of them (69%) had received an allogeneic transplant and thirteen (31%) an autologous transplant. Their median age was 7 years (range; 1-18). The most frequent reason for admission was respiratory failure (37 cases, 88%). The overall probability of developing complications requiring PICU admission was 21.2% (33.5% for allogeneic transplantation and 10.1% for patients receiving autologous grafts, p=0.0002). On univariate analysis, only the type of transplantation was significantly associated with PICU admission (allogeneic vs autologous RR 1.92, 95% CI: 1.46-2.53)(p = 0.0001). In allogeneic transplants, only the underlying disease (non-malignant) and the status of disease at transplantation within malignant diseases (advanced phase) were pretransplant variables associated with PICU admission. Post-transplantation risk factors were presence of graft-versus-host disease (GvHD) (p = 0.046) and its grade (II-IV) (p = 0.002), as well as the presence of multiorgan dysfunction during the early post-infusion phase especially when the lung was the first failing organ (p = 0.0001). However, on multivariate analysis, only severe GvHD was statistically significant. In the autologous transplantation group, the underlying disease (solid tumor, p = 0.07) and status at transplantation (advanced phase, p = 0.0029) were the only risk factors. In the post-transplant phase, patients who develop multiorgan dysfunction during the neutropenic period and those with engraftment syndrome had an increased risk of requiring critical care. The overall event-free survival (EFS) at 3 years was 15.3%, (18.4% for autologous transplant recipients and 13.7% for those receiving an allogeneic graft, p = 0.4). Using a Cox regression model, multiorgan failure (MOF) present at admission was the only variable that had a negative impact on EFS (4.28% vs 35.71% for patients with no MOF, p = 0.016). INTERPRETATION AND CONCLUSIONS: Despite high mortality, intensive care support can be beneficial for pediatric patients with life-threatening complications following HSCT. However, for patients with multiorgan failure involving the lungs, admission to the PICU should be avoided. PMID- 11869943 TI - Allogeneic transplantation after reduced conditioning in high risk patients is complicated by a high incidence of acute and chronic graft-versus-host disease. AB - Background and Objectives. We studied the toxicity and efficacy of reduced intensity conditioning followed by allogeneic stem cell transplantation in 50 patients over 50 years old or with relative contraindications against myeloablative regimens. Diagnoses were chronic myeloid leukemia (n=15), acute myeloid leukemia (n=9), myelodysplastic syndromes (n=9), lymphoma (n=11) and refractory solid tumors (n=6). Design and Methods. Donors were identical siblings (n=25), non-identical family members (n=6) and unrelated volunteers (n=19). Peripheral blood stem cells (n=36) or bone marrow (n=14) were transplanted. The conditioning regimen consisted of fludarabine 180 mg/m(2), busulphan 8 mg/kg and rabbit antithymocyte globulin 40 mg/kg (Fresenius). Graft-versus-host disease (GVHD) prophylaxis was carried out with cyclosporin A (CSA) alone (n=17) or in combination with methotrexate (n=18) or mycophenolate mofetil (n=15). Results. Neutrophil counts >0.5/nL and platelet counts > 20/nL were reached after 17 (range 0-66) and 19 days (range 0-111), respectively. Three graft failures occurred. Fever lasted for a median of 2 days (range 0-15). Six patients developed veno-occlusive disease of the liver. Acute GVHD grade II-IV occurred in 47% of the patients and chronic GVHD in 46%. The 1-year overall survival probability was 44% (95% CI: 30-58%). GVHD-related complications were a major cause of the probability of 1-year non-relapse mortality of 31% (95% CI: 16-46%). Interpretation and Conclusions. In conclusion, the regimen itself can be carried out safely in patients with relative contraindications against myeloablative conditioning. However, GVHD causes significant non-relapse mortality in high risk patients. PMID- 11869944 TI - Acute myeloid leukemia with t(8;21)/AML1/ETO: a distinct biological and clinical entity. AB - BACKGROUND AND OBJECTIVES: Recent investigations in acute myeloid leukemia (AML) have clearly demonstrated that specific karyotypic abnormalities result in distinct biological and clinical entities. We focus on recent advances on biology and treatment of AML with t(8;21). DATA SOURCES AND METHODS: The information presented here derives from literature data and experience of the authors. The most relevant studies are critically analyzed and discussed. STATE OF ART: Peculiar molecular, morphologic, immunophenotypic and epidemiologic findings of AML with t(8;21) as well as current methods for the evaluation of minimal residual disease are presented. Results from current therapeutic options including consolidation chemotherapy or transplantation procedures are critically reviewed. PERSPECTIVES: Innovative therapeutic approaches based on risk-adapted, patient-oriented approaches would be possible in this AML subtype, provided that answers to many unresolved questions are given. PMID- 11869945 TI - Efficacy of novel culture environments on the ex vivo expansion kinetics of cord blood progenitor cells. AB - We demonstrate the efficacy of two cytokine-rich supernatants, one from peripheral blood cell cultures stimulated with anti-CD3 and the other from monocyte-conditioned media, in enhancing the rapid (within 6 days) expansion and clonogenicity of CD34+, progenitor cells. We, thereby,increased the generation of mostly myeloid precursors able to support stem cell transplants. PMID- 11869946 TI - Combined therapy with amifostine plus erythropoietin for the treatment of myelodysplastic syndromes. AB - Twelve patients with myelodysplasia were treated with amifostine plus recombinant human erythropoietin (rHuEpo) for 6 weeks. A complete erythroid response was obtained in 2/12(16.6%) and a partial response in 4/12 (33.3%). Two of 8 patients with a platelet count < 100 x 10(9)/L had a complete response, as did 3/9 with a neutrophil count < 1.5 x 10(9)/L. Compared to rHuEpo or amifostine used as single agents, their combination did not offer substantial advantages. PMID- 11869947 TI - Hemoglobin F synthesis is not restricted to fetal erythropoietic organs during extramedullary hematopoiesis. AB - We investigated whether the anatomic distribution of hematopoietic cells determines the type of hemoglobin produced in patients with extramedullary hematopoiesis (EMH). Fetal hemoglobin (HbF) production is not restricted to fetal erythropoietic organs during EMH. A shift of erythropoiesis to fetal hematopoietic organs in EMH does not necessarily induce HbF synthesis in adulthood. PMID- 11869948 TI - Altered constitutive and activation-induced expression of CD95 by B- and T-cells in B-cell chronic lymphocytic leukemia. AB - Expression of CD95, a molecule involved in activation-induced cell death (AICD), might contribute to explain accumulation of leukemic B-cells and functional impairment of T-cells in B-cell chronic lymphocytic leukemia (B-CLL). There-fore, we compared constitutive and activation-induced expression of CD95 and CD69 by B- and T-cells in CLL patients and in healthy donors. PMID- 11869950 TI - Genotypic heterogeneity may explain phenotypic variations in inherited factor VII deficiency. AB - Inherited factor VIl (FVII) deficiency is a rare autosomal recessive coagulation disorder characterized by a wide genet-ic heterogeneity and a poor relationship between FVII activity (FVII:C) levels and severity of the hemorrhagic diathesis. Given both the rarity and the heterogeneity of this disorder,genotype-phenotype relationships are difficult to clarify. The analysis of three FVII-deficient patients enabled us to offer some explanations. PMID- 11869949 TI - Advantages of using thalidomide for the management of refractory myeloma patients. AB - A group of 11 heavily pretreated patients receiving low-dose thalidomide was compared with a similar group of 10 patients with refractory myeloma treated with a convention-al oral chemotherapy. This study shows that thalidomide is not only effective in controlling the neoplastic clone but more-over, thanks to its low toxicity, allows out-patient management of these subjects. PMID- 11869951 TI - A novel case of immunodeficiency, centromeric instability, and facial anomalies (the ICF syndrome): immunologic and cytogenetic studies. AB - The immunodeficiency, centromeric instability, and facial anomalies (ICF) syndrome is characterized by hypogammaglobulinemia and recurrent bacterial infections. Here we report a novel case of ICF syndrome with hypogammaglobulinemia and an inverted CD4/CD8 ratio. Cytogenetically abnormal cells,that were identified in both CD4+ and CD4- peripheral blood lymphocytes, retained their ability to proliferate in vitro following polyclonal stimulation. A primitive defect of B-cell differentiation was detected. PMID- 11869952 TI - Second allogeneic hematopoietic stem cell transplantation in hematologic malignancies in children: long-term results of a multicenter study of the Spanish Working Party for Bone Marrow Transplantation in Children (GETMON). AB - Twenty-four children with acute leukemia (21) or chronic myeloid leukemia (3) who relapsed after a first hematopoietic stem cell transplantation (HSCT) underwent a second allogeneic HSCT. Sixteen patients died from relapse or transplant related causes and 8 are alive and disease-free with a probability of event-free survival at 5 years of 32%. These results show that this procedure offers a chance to a subset of these patients. PMID- 11869953 TI - Tandem high-dose therapy followed by autologous stem-cell transplantation for refractory or relapsed high grade non-Hodgkin's lymphoma with poor prognosis factors: a prospective pilot study. AB - We conducted a tandem autologous stem cell transplantation procedure for patients with relapsed or refractory high-grade non-Hodgkin's lymphoma (HGNHL) or with transformation of indolent lymphoma (n=15). These patients had poor prognosis factors. The procedure was well tolerated, ten patients were in complete remission. Overall survival rate is 67%. PMID- 11869955 TI - Extramedullary myeloid cell tumor/granulocytic sarcoma with predilection for serosal surfaces. PMID- 11869954 TI - ATP downregulation in mononuclear cells from children with graft-versus-host disease following extracorporeal photochemotherapy. AB - Graft-versus-host disease (GvHD) is a frequent and major complication of allogeneic bone marrow transplantation(BMT). Acute GvHD occurs in 40% to 50% of allogeneic BMT recipients; chronic GvHD can be observed in 30% to 60% of long term survivors. PMID- 11869956 TI - Keratoderma blenorrhagica, balanitis and uveitis after HLA-B15 mismatched bone marrow transplantation. PMID- 11869957 TI - Hepatosplenic gamma delta T-cell lymphoma with a novel cytogenetic alteration and cerebrospinal fluid infiltration: biological and clinical features. PMID- 11869958 TI - Bone marrow of a patient with acute non-lymphoblastic leukaemia associated with the t(1;7)(p11;p11) chromosomal translocation. PMID- 11869959 TI - Blast crisis merely lymph nodal of chronic myeloid leukemia. PMID- 11869960 TI - Cuban experience in the treatment of acute promyelocytic leukemia with ALL transretinoic acid followed by intensive chemotherapy. PMID- 11869961 TI - What is the dose of STI-571 needed to induce a molecular remission in chronic myeloid leukemia? PMID- 11869962 TI - High expression of apoptosis-regulating proteins in Burkitt's lymphomas. PMID- 11869963 TI - Further remarks on the expression of CD20 in classical Hodgkin's lymphomas. PMID- 11869964 TI - Efficacy of thalidomide in the treatment of myelodysplastic syndromes. PMID- 11869965 TI - Spreading your way to lower cholesterol? PMID- 11869966 TI - Keeping heart failure from putting you in the hospital. PMID- 11869967 TI - Growing around blocked arteries. PMID- 11869968 TI - New angina drugs on the horizon? PMID- 11869970 TI - Making blood pressure movies. PMID- 11869969 TI - Cutting salt better than exercise at controlling blood pressure. PMID- 11869971 TI - Aggressive therapy for older hearts. PMID- 11869972 TI - Ask the doctor. Does grapefruit pectin powder unclog arteries? Are there any side effects? PMID- 11869973 TI - Ask the doctor. Because I have a pacemaker, I've always avoided going through the big scanners at the airport and have gone around to the person with the hand-held device. As airport security gets tighter, I'm worried that my reluctance to go through metal detectors will single me out. Is it safe for me to go through them? PMID- 11869975 TI - Depression in children - Part I. PMID- 11869976 TI - EMDR. PMID- 11869977 TI - Depression and anxiety: deficiencies in treatment. PMID- 11869978 TI - Placebos and suicide. PMID- 11869979 TI - The traumatized heart. PMID- 11869980 TI - Viagra for depression. PMID- 11869981 TI - Emotional expression in criminals. PMID- 11869985 TI - The new margarines: can they help your heart? PMID- 11869986 TI - The swollen or painful scrotum. PMID- 11869987 TI - Declining deaths from prostate cancer. PMID- 11869988 TI - On Call. I am a subscriber to Harvard Men's Health Watch, but I am writing about my wife, not myself. She's only 58 and she's always been healthy, but over the past year she's been bothered by shaking and trembling in her hands, particularly when she's pouring tea or eating dinner. Her doctor gave her a blood pressure pill, but she's never had high blood pressure. Does she have Parkinson's? Should I take her to a specialist? PMID- 11869989 TI - Taking stock of antioxidants. PMID- 11869990 TI - Arthritis. Hip replacement. PMID- 11869991 TI - Raising HDL cholesterol: a worthwhile goal? PMID- 11869992 TI - Stroke. Carotid artery narrowings: surgery versus stents. PMID- 11869993 TI - By the way doctor... I'm 72 and was recently told that I am losing a bit of my sight because of macular degeneration. My doctor assured me that it was progressing very slowly, but also said there wasn't really anything to do about it. But I read that zinc or vitamins might help. Should I be taking them? PMID- 11869994 TI - Considering disruptive behaviors. PMID- 11869996 TI - Personality pathology and treatment outcome in major depression: a review. AB - OBJECTIVE: A longstanding belief among many clinicians is that patients with depression and comorbid personality pathology have a worse response to standard depression treatment. This presents potentially significant treatment implications, since personality pathology in depressed patients appears to be common. METHOD: PsycINFO and MEDLINE were systematically searched for studies relating personality to treatment outcome. Over 50 studies were obtained and grouped according to the method used to assess personality pathology. RESULTS: High neuroticism scores generally predicted worse outcome, especially over long term follow-up. Tridimensional Personality Questionnaire scores did not have a consistent relationship to treatment outcome despite some promising initial findings. Most studies involved patients with comorbid personality disorders; these studies produced conflicting results. Other measures of personality pathology produced an array of findings ranging from a moderately worse outcome to no difference. CONCLUSIONS: Whether or not personality pathology significantly worsens outcome in patients with major depression appears to depend on study design, since the rate of personality pathology varies markedly depending on how it is measured. In addition, depressed patients with personality pathology appear less likely to receive adequate treatment in uncontrolled studies. Finally, studies rarely control for depression characteristics (e.g., chronicity, severity) that may influence outcome and be related to personality pathology. Overall, the best-designed studies reported the least effect of personality pathology on depression treatment outcome. Clinically, this suggests that comorbid personality pathology should not be seen as an impediment to good treatment response. PMID- 11869995 TI - Can a valid diagnosis of disruptive behavior disorder be made in preschool children? AB - OBJECTIVE: Disruptive behavior problems are the most common reason preschool children come to mental health clinics. Yet consensus on the conceptualization and measurement of such problems in young children is lacking. DSM-IV is the most widely used nosologic system for children, but the majority of the validation sample consisted of school-age children and adolescents. It is debatable whether behavioral problems in young children should be considered within a diagnostic framework at all, since normative behavioral disruption occurs during the preschool period. Developing valid methods for assessing child behavior problems across development is critical for etiologic and prevention research. METHOD: The authors compare different approaches to conceptualizing disruptive behavior in young children, review evidence for the construct validity of DSM-based oppositional defiant disorder and conduct disorder in preschool children, and outline an agenda for future research. RESULTS AND CONCLUSIONS: Typical and atypical behavior problems can be differentiated in preschool children, and the DSM framework, with some modification to address the child's developmental level, appears to be a valid method for identifying preschool children with disruptive behavior that is impairing. Empirical investigation is needed to standardize modification of existing assessment tools so that they can be used with preschool children and to develop more clinically sensitive methods for using observational data in assessment and for establishing the child's level of development. PMID- 11869997 TI - The human genome: detecting chromosomal deletions: Angelman and Prader-Willi syndromes. PMID- 11869999 TI - Johann Bernhard Aloys von Gudden, 1824-1886. PMID- 11869998 TI - Consultation-liaison psychiatry: a longitudinal and integrated approach. PMID- 11870000 TI - Should the DSM-IV diagnostic criteria for conduct disorder consider social context? AB - OBJECTIVE: The text of the DSM-IV states that a diagnosis of conduct disorder should be made only if symptoms are caused by an internal psychological dysfunction and not if symptoms are a reaction to a negative environment. However, the DSM-IV diagnostic criteria are purely behavioral and ignore this exclusion. This study empirically evaluated which approach--the text's negative environment exclusion or the purely behavioral criteria--is more consistent with clinicians' intuitive judgments about whether a disorder is present, whether professional help is needed, and whether the problem is likely to continue. METHOD: Clinically experienced psychology and social work graduate students were presented with three variations of vignettes describing youths whose behavior satisfied the DSM-IV criteria for conduct disorder. The three variations presented symptoms only, symptoms caused by internal dysfunction, and symptoms caused by reactions to a negative environment. The clinicians rated their level of agreement that the youth described in the vignette had a disorder, needed professional mental health help, and had a problem that was likely to continue into adulthood. RESULTS: Youths with symptoms caused by internal dysfunction were judged to have a disorder, and those with a reaction to a negative environment not to have a disorder. The difference was not explained by the clinicians' judgments of the youths' need for professional help or the expected duration of symptoms. CONCLUSIONS: The clinicians' judgments supported the validity of the DSM-IV's textual claim that a diagnosis of conduct disorder is valid only when symptoms are due to an internal dysfunction. PMID- 11870001 TI - Childhood adversity and the endogenous versus nonendogenous distinction in women with major depression. AB - OBJECTIVE: Previous research has reported an association between childhood adversity and the nonendogenous subtype of depression. The present study sought to address this association by 1) investigating the relationship between depression subtype and different levels of childhood adversity and 2) examining the relative contribution of different types of childhood adversity to depression subtypes. METHOD: The authors conducted an investigator-based assessment of childhood adversity in a community group of 76 depressed women, 31 of whom met Research Diagnostic Criteria for endogenous depression. RESULTS: In contrast to previous studies, severe physical abuse, sexual abuse, antipathy, and neglect were significantly and preferentially associated with endogenous depression, as were both high and lax levels of supervision and discipline. In multivariate analyses, severe sexual abuse and a composite variable comprising antipathy and neglect were most strongly associated with endogenous depression. CONCLUSIONS: Implications of these findings in the context of the existing literature are discussed, and potential mediators of the relationship between childhood adversity and endogenous depression are presented as avenues for future research. PMID- 11870002 TI - Childhood adversities associated with risk for eating disorders or weight problems during adolescence or early adulthood. AB - OBJECTIVE: A community-based prospective longitudinal study was conducted to investigate the association between childhood adversities and problems with eating or weight during adolescence and early adulthood. METHOD: A community based sample of 782 mothers and their offspring were interviewed during the childhood, adolescence, and early adulthood of the offspring. Childhood maltreatment, eating problems, environmental risk factors, temperament, maladaptive parental behavior, and parental psychopathology were assessed during childhood and adolescence. Eating disorders and problems with eating or weight in the offspring were assessed during adolescence and early adulthood. RESULTS: A wide range of childhood adversities were associated with elevated risk for eating disorders and problems with eating or weight during adolescence and early adulthood after the effects of age, childhood eating problems, difficult childhood temperament, parental psychopathology, and co-occurring childhood adversities were controlled statistically. Numerous unique associations were found between specific childhood adversities and specific types of problems with eating or weight, and different patterns of association were obtained among the male and female subjects. Maladaptive paternal behavior was uniquely associated with risk for eating disorders in offspring after the effects of maladaptive maternal behavior, childhood maltreatment, and other co-occurring childhood adversities were controlled statistically. CONCLUSIONS: Childhood adversities may contribute to greater risk for the development of eating disorders and problems with eating and weight that persist into early adulthood. Maladaptive paternal behavior may play a particularly important role in the development of eating disorders in offspring. PMID- 11870003 TI - Pathways of self-reported problem behaviors from adolescence into adulthood. AB - OBJECTIVE: The authors determined the impact of different pathways of psychopathological development on adult outcome in subjects followed from ages 11 18 to ages 21-28. METHOD: Problem behaviors of subjects from a general population sample were assessed through the Youth Self-Report and the Young Adult Self Report given at four time points (1987, 1989, 1991, and 1997). In addition, DSM IV diagnoses, information pertaining to signs of maladjustment, and measures of social functioning were obtained at the last assessment. On the basis of the self report ratings, four contrasting developmental pathways of psychopathology were determined: persistent, decreasing, increasing, and consistently normal. RESULTS: Subjects whose overall level of psychopathology was persistent over time had a higher lifetime prevalence of DSM-IV diagnoses and a poorer general outcome in adulthood than did subjects whose level of psychopathology increased. Subjects whose level of psychopathology returned to normal after high levels of problems in adolescence were only slightly different in terms of outcome from subjects with consistently normal ratings. CONCLUSIONS: 1) People who showed high levels of problems in early adolescence but whose level of psychopathology diminished by adulthood seemed to be as healthy as people who never attained a serious level of psychopathology. 2) An ongoing devious pathway into adulthood had negative effects on many domains of functioning. These two findings are both powerful arguments for early intervention in adolescence. PMID- 11870004 TI - Self-injurious behavior in women with eating disorders. AB - OBJECTIVE: The authors assessed lifetime and 6-month occurrence and phenomenology of self-injurious behavior in patients with eating disorders. METHOD: Women (N=376) in inpatient treatment for an eating disorder (anorexia: N=119, bulimia: N=137, eating disorder not otherwise specified: N=120) were assessed for self injurious behavior and completed the Traumatic Life Events Questionnaire, the Dissociative Experience Scale, the Barratt Impulsiveness Scale, and the Yale Brown Obsessive Compulsive Scale. RESULTS: The lifetime rate of self-injurious behavior occurrence was 34.6%, with the highest rates found in subjects with eating disorder not otherwise specified (35.8%) and bulimia (34.3%); the 6-month rate of self-injurious behavior occurrence was 21.3%. Multivariate comparisons were computed for the factors of self-injurious behavior and diagnostic subgroup: self-injuring patients reported a significantly higher number of traumatic events, showed significantly higher dissociation scores, and exhibited significantly more obsessive-compulsive thoughts and behaviors. Bulimic patients showed significantly higher impulsivity scores. CONCLUSIONS: This study strongly supports the assumption that patients with eating disorders are at risk for self injurious behavior and points to the necessity of a routine screening for self injurious behavior as well as the development of a standardized questionnaire. Group comparisons point to the relevance of traumatic experiences and comorbid dissociative phenomenology. PMID- 11870005 TI - Latent structure of eating disorder symptoms: a factor analytic and taxometric investigation. AB - OBJECTIVE: The latent structure of eating disorder symptoms, as defined by DSM IV, was tested in a group of 341 women with and without an eating disorder diagnosis. METHOD: The study group consisted of 201 participants with a diagnosis of anorexia nervosa, bulimia nervosa, binge eating disorder, or eating disorder not otherwise specified; 24 comparison subjects who were obese but did not have an eating disorder diagnosis; and 116 normal-weight comparison subjects. The presence and severity of DSM-IV eating disorder symptoms was assessed with the semi-structured Interview for the Diagnosis of Eating Disorders-IV. The study group was randomly divided into two subgroups for factor analytic studies, and the data were subjected to exploratory and confirmatory factor analysis. Pilot taxometric analyses were used to examine whether the obtained factors represented true dimensions or latent discrete classes. RESULTS: In exploratory factor analyses with data from subgroup 1, three factors were found to account for 66% of the variance in eating disorder symptoms: binge eating, fear of fatness/compensatory behaviors, and drive for extreme thinness. Confirmatory factor analysis cross-validated this factor structure with data from subgroup 2. The eating disorder groups and comparison groups were found to differ on at least one of the three factors. The results of the taxometric analyses were inconsistent with a strictly dimensional model of eating disorders and suggested that some features may be dimensional whereas others may be taxonic (discrete). DISCUSSION: The eating disorders, as defined by DSM-IV, can be conceptualized as having three latent features. Taxometric tests found empirical support for conceptualizing bulimia nervosa and binge eating disorder as discrete syndromes. PMID- 11870007 TI - Greater loss of 5-HT(2A) receptors in midlife than in late life. AB - OBJECTIVE: Earlier work has shown markedly lower density of serotonin 2A (5 HT(2A)) receptors in elderly subjects than in young healthy subjects. In this study the authors used positron emission tomography (PET) and [(18)F]altanserin, a ligand with high affinity for the 5-HT(2A) receptor, to examine the relationship between 5-HT(2A) receptor density and age in more detail. METHOD: The 22 subjects ranged in age from 21 to 69 years (mean=43.4, SD=13.3) and were healthy comparison subjects in a study of depression. Regions of interest were determined on magnetic resonance images and were transferred to coregistered PET data. The data were derived from dynamic PET scanning and arterial sampling with resulting plasma activity data corrected for labeled metabolites. Compartmental modeling was used to estimate the radioligand distribution volume. By comparing the distribution volume (DV) of different regions to the cerebellum distribution volume, DV(ratio)-1, which is proportional to the binding potential, was calculated. RESULTS: The decrease in 5-HT(2A) binding was not linear but on average was approximately 17% per decade from age 20. The correlations between age and 5-HT(2A) DV(ratio)-1 were significant for the global measure and for the medial gyrus rectus, anterior cingulate, posterior medial prefrontal cortex, hippocampus, and occipital cortex. Most of the fall off in receptor binding occurred up through midlife, and there was less decrease in late life. There were no decreases in regional brain volumes of corresponding magnitudes. CONCLUSIONS: 5-HT(2A) receptor binding decreases dramatically in a variety of brain regions up through midlife. PMID- 11870006 TI - Higher expression of serotonin 5-HT(2A) receptors in the postmortem brains of teenage suicide victims. AB - OBJECTIVE: Abnormalities of serotonin (5-HT) receptor subtypes have been observed in the postmortem brains of adult suicide victims; however, their role in teenage suicide is unexplored. The authors examined whether 5-HT(2A) receptor subtypes are altered in the postmortem brains of teenage suicide victims. METHOD: Levels of 5-HT(2A) receptors were determined through examination of [(125)I] LSD binding, protein expression (by use of Western blotting with a specific 5-HT(2A) receptor antibody), and mRNA (by means of quantitative reverse transcription polymerase chain reaction) in the prefrontal cortex, hippocampus, and nucleus accumbens of 15 teenage suicide victims and 15 normal matched teenage subjects. The cellular localization of the 5-HT(2A) receptors was determined by means of gold immunolabeling. RESULTS: The authors observed significantly higher [(125)I]LSD binding in the prefrontal cortex and greater protein expression and mRNA levels in the prefrontal cortex and hippocampus but not in the nucleus accumbens of suicide victims, compared with normal subjects. Greater protein expression was localized on pyramidal cells in cortical layer V but not in other cortical layers or in the surrounding neuropil of the prefrontal cortex of teenage suicide victims. CONCLUSIONS: The evidence indicates higher levels of 5 HT(2A) receptor, protein, and mRNA expression in the prefrontal cortex and hippocampus, which have been implicated in emotion, stress, and cognition. There was no higher level in the nucleus accumbens, which has been implicated in drug dependence and craving. Our findings suggest that a higher level of 5-HT(2A) receptors may be one of the neurobiological abnormalities associated with teenage suicide. PMID- 11870008 TI - Differential evolution of cognitive impairment in nondemented older persons: results from the Kungsholmen Project. AB - OBJECTIVE: This study was a prospective, population-based examination of the evolution of cognitive impairment, no dementia (CIND). METHOD: Subjects 75 years old or older living in Stockholm were assessed at baseline and 3 and 6 years later. The severity of CIND was based on age- and education-specific norms on the Mini-Mental State Examination and was classified as mild (N=212), moderate (N=96), or severe (N=57). Mortality, progression to dementia (DSM-III-R), cognitive stability, and cognitive improvement were studied as main outcomes. RESULTS: Of the individuals with mild CIND, 63 (34%) died, 65 (35%) progressed to dementia, 21 (11%) remained stable, and 46 (25%) improved between baseline and first follow-up. The relative risks of progressing to dementia by first follow-up in the subjects with mild, moderate, and severe CIND were 3.6, 5.4, and 7.0, respectively. The relative risk of death decreased with increasing severity of impairment. Individuals who improved at first follow-up did not have a significantly higher risk of later progressing to dementia than subjects who had never been impaired (relative risk=1.4). The absence of a subjective memory complaint predicted improvement (odds ratio=5.4). CONCLUSIONS: CIND is a heterogeneous condition: similar proportions of subjects progress to dementia, death, and cognitive improvement over 3 years. There is no increased future risk of progressing to dementia in CIND subjects who improve during that period. PMID- 11870009 TI - Are cognitive impairments associated with sensitivity to stress in schizophrenia? An experience sampling study. AB - OBJECTIVE: Patients with a diagnosis of schizophrenia display cognitive impairments and abnormal sensitivity to stress. However, little is known about the relationship between these two endophenotypes. METHOD: Neuropsychological tests were administered to 42 patients with schizophrenia or other psychosis to assess cognitive functioning, and the experience sampling method (a structured diary technique assessing current context and mood in daily life) was used to assess 1) appraised subjective stress related to daily events and activities and 2) emotional reaction to these daily life stressors. RESULTS: Multilevel random regression analyses showed that in some instances, cognitive functioning did not alter emotional reaction to stress. In other instances, an inverse relationship was found, indicating that a better performance on neuropsychological tests was related to greater emotional reaction to stress. CONCLUSIONS: The results indicate that emotional reaction to stress in the daily lives of patients with schizophrenia may not be a consequence of cognitive impairments and that the two mechanisms may act through different pathways. Such pathways may be related to the extremes of clinical outcome that have been observed in schizophrenia: an episodic, reactive, good outcome and a more chronic form characterized by high levels of negative symptoms and cognitive impairments. PMID- 11870010 TI - Mental disorder in elderly suicides: a case-control study. AB - OBJECTIVE: The authors' goal was to study the importance of different psychiatric disorders in relation to suicide in individuals 65 years old or older. METHOD: The psychological autopsy approach was used to study 85 cases of suicide among subjects who were 65 years old or older; 153 living comparison subjects from the same age group who were randomly selected from the tax register were interviewed face-to-face. Retrospective axis I diagnoses were made according to DSM-IV on the basis of interview data and medical records. RESULTS: Ninety-seven percent of the suicide victims fulfilled criteria for at least one DSM-IV axis I diagnosis, compared with 18% of the living comparison subjects. Recurrent major depressive disorder was a very strong risk factor for suicide, as was substance use disorder. An elevated risk was also associated with minor depressive disorder, dysthymic disorder, psychotic disorder, single-episode major depressive disorder, and anxiety disorder. Comorbid axis I disorders were observed in 15 (38%) of the 39 elderly subjects with major depressive disorder who had committed suicide. CONCLUSIONS: Although recurrent major depressive disorder was the mental disorder most strongly associated with suicide, the findings of this study suggest that elderly individuals who commit suicide represent a heterogeneous group with regard to mental disorders, implying a need for differentiated prevention strategies. PMID- 11870011 TI - Low testosterone levels in elderly men with dysthymic disorder. AB - OBJECTIVE: A decline in hypothalamic-pituitary-gonadal (HPG) axis function is often seen in elderly men, and dysthymic disorder is common. Symptoms of both HPG axis hypofunction and dysthymic disorder include dysphoria, fatigue, and low libido. The authors compared total testosterone levels in three groups of elderly men. METHOD: Total testosterone levels were measured in subjects who met DSM-IV criteria for major depressive disorder (N=13) or dysthymic disorder (N=32) and a comparison group (N=175) who had participated in an epidemiological study of male aging and had scored below the median on the Center for Epidemiologic Studies Depression Scale, a well-validated, self-report depression symptom inventory. RESULTS: There were no differences among the three groups in measured demographic variables, including age and weight. Median testosterone levels varied for those with dysthymic disorder (295 ng/dl), major depressive disorder (425 ng/dl), and no depression (423 ng/dl). A test for differences in central tendency showed a statistically significant difference among the three groups. Post hoc pairwise comparisons revealed statistically significant differences between those with dysthymic disorder and those with major depressive disorder and no depression. CONCLUSIONS: Total testosterone levels were lower in elderly men with dysthymic disorder than in men with major depressive disorder and men without depressive symptoms. Dysthymic disorder in elderly men may be related to HPG axis hypofunction. PMID- 11870012 TI - Comparison of citalopram, perphenazine, and placebo for the acute treatment of psychosis and behavioral disturbances in hospitalized, demented patients. AB - OBJECTIVE: Until recently, conventional antipsychotics were the standard pharmacotherapy for psychosis and behavioral disturbances associated with dementia. This double-blind, placebo-controlled study compared the acute efficacy of the selective serotonin reuptake inhibitor citalopram and the neuroleptic perphenazine with placebo for the treatment of psychosis and behavioral disturbances in nondepressed patients with dementia. METHOD: Eighty-five hospitalized patients with at least one moderate to severe target symptom (aggression, agitation, hostility, suspiciousness, hallucinations, or delusions) were randomly assigned to receive either citalopram, perphenazine, or placebo under double-blind conditions for up to 17 days. RESULTS: Patients treated with citalopram or perphenazine showed statistically significant improvement on several Neurobehavioral Rating Scale factor scores. Compared to those receiving placebo, only patients treated with citalopram showed significantly greater improvement in their total Neurobehavioral Rating Scale score as well as in the scores for the agitation/aggression and lability/tension factors. Side effect scores were similar among the three treatment groups. CONCLUSIONS: Citalopram was found to be more efficacious than placebo in the short-term hospital treatment of psychotic symptoms and behavioral disturbances in nondepressed, demented patients. PMID- 11870013 TI - Combined pharmacotherapy and psychotherapy as maintenance treatment for late-life depression: effects on social adjustment. AB - OBJECTIVE: This study examined the effects of maintenance treatment for late-life depression on social adjustment. The authors hypothesized that elderly patients recovering from depression would have better social adjustment with medication and interpersonal psychotherapy than with medication or psychotherapy alone. METHOD: Patients aged 60 and older recovering from recurrent major depression were randomly assigned to one of four treatments: nortriptyline and interpersonal psychotherapy, nortriptyline and clinic visits, placebo and psychotherapy, or placebo and clinic visits. The Social Adjustment Scale was administered every 3 months until illness recurrence. Combined treatment was compared to monotherapy on scores over 1 year among patients who remained in recovery (N=49). RESULTS: Patients receiving nortriptyline and interpersonal psychotherapy maintained social adjustment, which declined in those receiving monotherapy. CONCLUSIONS: Treatment of late-life depression with nortriptyline and psychotherapy is more likely to maintain social adjustment than treatment with either alone. Combination therapy improves not only duration but quality of wellness. PMID- 11870014 TI - Are subjects in pharmacological treatment trials of depression representative of patients in routine clinical practice? AB - OBJECTIVE: The methods used to evaluate the efficacy of antidepressants differ from treatment for depression in routine clinical practice. The rigorous inclusion/exclusion criteria used to select subjects for participation in efficacy studies potentially limit the generalizability of these trials' results. It is unknown how much impact these criteria have on the representativeness of subjects in efficacy trials. This study estimated the proportion of depressed patients treated in routine clinical practice who would meet standard inclusion/exclusion criteria for an efficacy trial. METHOD: A total of 803 individuals, aged 16--65 years, who were seen at intake at an outpatient practice underwent a thorough diagnostic evaluation, including the administration of semistructured diagnostic interviews; 346 patients had current major depression. Common inclusion/exclusion criteria used in efficacy studies of antidepressants were applied to the depressed patients to determine how many would have qualified for an efficacy trial. RESULTS: Approximately one-sixth of the 346 depressed patients would have been excluded from an efficacy trial because they had a bipolar or psychotic subtype of depression. The presence of a comorbid anxiety or substance use disorder, insufficient severity of depressive symptoms, or current suicidal ideation would have excluded 86.0% (N=252) of the remaining 293 outpatients with nonpsychotic unipolar major depressive disorder from an antidepressant efficacy trial. CONCLUSIONS: Subjects treated in antidepressant trials represent a minority of patients treated for major depression in routine clinical practice. These results show that antidepressant efficacy trials tend to evaluate a subset of depressed individuals with a specific clinical profile. PMID- 11870015 TI - Combined Analysis of CSF Tau Levels and [(123)I]Iodoamphetamine SPECT in Mild Cognitive Impairment: Implications for a Novel Predictor of Alzheimer's Disease. AB - OBJECTIVE: The aim of this study was to establish an objective and reliable index to predict the development of Alzheimer's disease in a large pool of elderly patients with mild cognitive impairment. METHOD: Twenty-three patients with probable Alzheimer's disease, 22 patients with mild cognitive impairment who eventually developed Alzheimer's disease, eight patients with mild cognitive impairment who did not develop dementia, and 19 cognitively normal subjects were included in the study. The authors constructed a new diagnostic index, the CSF CBF index, based on CSF tau levels divided by regional cerebral blood flow (CBF) in the posterior cingulate cortex. RESULTS: Receiver operating characteristic analysis showed that applying a cutoff value for the CSF-CBF index of 296.0 achieved a sensitivity of 88.5% and a specificity of 90.0% in discriminating mild cognitive impairment that progressed to Alzheimer's disease from mild cognitive impairment that did not progress to Alzheimer's disease. CONCLUSIONS: The CSF-CBF index is useful in predicting Alzheimer's disease in subjects with mild cognitive impairment. PMID- 11870016 TI - Addition of omega-3 fatty acid to maintenance medication treatment for recurrent unipolar depressive disorder. AB - OBJECTIVE: Studies have reported that countries with high rates of fish oil consumption have low rates of depressive disorder. The authors studied a specific omega-3 fatty acid, the ethyl ester of eicosapentaenoic acid (E-EPA), as an adjunct to treatment for depressive episodes occurring in patients with recurrent unipolar depressive disorder who were receiving maintenance antidepressant therapy. METHOD: Twenty patients with a current diagnosis of major depressive disorder participated in a 4-week, parallel-group, double-blind addition of either placebo or E-EPA to ongoing antidepressant therapy. Seventeen of the patients were women, and three were men. RESULTS: Highly significant benefits of the addition of the omega-3 fatty acid compared with placebo were found by week 3 of treatment. CONCLUSIONS: It is not possible to distinguish whether E-EPA augments antidepressant action in the manner of lithium or has independent antidepressant properties of its own. PMID- 11870017 TI - Placebo-controlled trial of D-cycloserine added to conventional neuroleptics, olanzapine, or risperidone in schizophrenia. AB - OBJECTIVE: The authors investigated the clinical effects of D-cycloserine when added to treatment with conventional neuroleptics, olanzapine, or risperidone for treatment-resistant schizophrenia. METHOD: Twenty-four patients participated in a double-blind, placebo-controlled, 6-week crossover trial with D-cycloserine, 50 mg/day, added to their fixed dose of antipsychotic medication. Clinical ratings were performed every 2 weeks. RESULTS: D-Cycloserine treatment was well tolerated and resulted in a significant reduction in negative symptoms (mean=15%). The degree of improvement did not differ between patients treated with conventional neuroleptics and those treated with olanzapine or risperidone. CONCLUSIONS: These data support the efficacy of the addition of 50 mg/day of D-cycloserine to treatment with conventional neuroleptics and suggest that therapeutic benefits may also be attained when D-cycloserine is added to olanzapine or risperidone. PMID- 11870018 TI - Neuropsychological function in children with maltreatment-related posttraumatic stress disorder. AB - OBJECTIVE: Studies in adults have reported changes in concentration, learning, and memory in individuals with posttraumatic stress disorder (PTSD). However, there are few studies of cognitive function in children with PTSD. The goal of the current study was to evaluate cognition in children with PTSD. METHOD: The cognitive status of 14 pediatric psychiatric outpatients with maltreatment related PTSD and 15 sociodemographically similar children who were healthy and had not been maltreated was examined. Neuropsychological instruments measured language, attention, abstract reasoning/executive function, learning and memory, visual-spatial processing, and psychomotor function. RESULTS: The children with PTSD performed more poorly on measures of attention and abstract reasoning/executive function. CONCLUSIONS: Although based on a small number of subjects, these results support cognitive differences between children with and without maltreatment-related PTSD. PMID- 11870019 TI - Pilot study of spirituality and mental health in twins. AB - OBJECTIVE: The goal of this study was to investigate associations between empirically defined dimensions of spirituality, personality variables, and psychiatric disorders in Vietnam era veterans. METHOD: One hundred pairs of male twins from the Vietnam Era Twin Registry were administered the self-report Spiritual Well-Being Scale and a pilot Index of Spiritual Involvement. Correlation analyses were supplemented with regression analyses that examined the relative influence of genetic and environmental factors on aspects of spirituality. RESULTS: Existential well-being was significantly associated with seven of 11 dimensions of personality and was significantly negatively associated with alcohol abuse or dependence and with two of three clusters of personality disorder symptoms. Associations between mental health variables and religious well-being or spiritual involvement were much more limited. CONCLUSIONS: Useful distinctions can be made between major dimensions of spirituality in studies of spirituality, religious coping, and mental health. PMID- 11870020 TI - Personality factors associated with dissociation: temperament, defenses, and cognitive schemata. AB - OBJECTIVE: The purpose of this study was to investigate temperamental, psychodynamic, and cognitive factors associated with dissociation. METHOD: Fifty three subjects with DSM-IV-defined depersonalization disorder and 22 healthy comparison subjects were administered the Dissociative Experiences Scale, the Tridimensional Personality Questionnaire, the Defense Style Questionnaire, and the Schema Questionnaire. RESULTS: Subjects with depersonalization disorder demonstrated significantly greater harm-avoidant temperament, immature defenses, and over-connection and disconnection cognitive schemata than comparison subjects. Within the group of subjects with depersonalization disorder, dissociation scores significantly correlated with the same variables. CONCLUSIONS: Particular personality factors may render individuals more vulnerable to dissociative symptoms. Risk factors associated with dissociative disorders merit further study. PMID- 11870021 TI - Orlistat misuse in bulimia nervosa. PMID- 11870022 TI - Desipramine toxicity with terbinafine. PMID- 11870023 TI - Olanzapine for violent schizophrenia and Klinefelter syndrome. PMID- 11870024 TI - Body dysmorphic disorder triggered by medical illness? PMID- 11870025 TI - Transcranial magnetic stimulation in schizophrenia. PMID- 11870026 TI - Clozapine and pericarditis with pericardial effusion. PMID- 11870027 TI - Origins of dreaming. PMID- 11870028 TI - Origins of dreaming. PMID- 11870030 TI - Meta-analysis and psychiatric genetics. PMID- 11870032 TI - Sweet taste preference and alcohol dependence. PMID- 11870033 TI - Shakespeare and successful aging. PMID- 11870035 TI - Amnesia and neonaticide. PMID- 11870036 TI - Sweet taste preference and alcohol dependence. PMID- 11870037 TI - Seasonal fluctuation in schizophrenia. PMID- 11870056 TI - Identification of amplified restriction fragment length polymorphism markers linked to genes controlling boar sperm viability following cryopreservation. AB - This study investigated two hypotheses: 1) that consistent between-boar variation in frozen semen quality exists and is genetically determined, and 2) molecular markers linked to genes controlling semen freezability can be identified using amplified restriction fragment length polymorphism (AFLP) technology. Five ejaculates were collected from each of 129 boars. Semen was diluted into a commercial freezing buffer (700 mOsm/kg, 3% glycerol) and five straws (0.5 ml) per ejaculate were cryopreserved (to -5 degrees C at 6 degrees C/min, then -5 degrees C to -80 degrees C at 40 degrees C/min). Semen was assessed for percentage of motile cells, motility characteristics (computer-aided semen analysis; CASA), plasma membrane integrity (SYBR-14 positive), and acrosome integrity (positive for fluorescein-labeled peanut agglutinin; PNA). Consistent between-boar variability was detected for postthaw sperm motility (P < 0.01), membrane integrity (P < 0.01), acrosome integrity (P < 0.01), and all CASA characteristics (P < 0.05). There was no significant difference between ejaculates (P > 0.05) or straws (P > 0.05) for any viability assessment. Multivariate pattern analysis of the viability data set highlighted three groups of boars producing spermatozoa with poor, average, and good postthaw recovery (42, 63, and 24 boars, respectively). DNA from Large White boars (n = 22) previously classified as good and poor freezers was screened for AFLP markers. Twenty-eight polymerase chain reaction primer combinations generated 2182 restriction fragment bands, of which 421 were polymorphic. Sixteen candidate genetic markers (P < 0.005) were identified by comparing the AFLP profile with semen freezability using logistic regression analysis. These findings support the hypothesis that there is a genetic basis for variation in postthaw semen quality between individuals, and that AFLP technology may be able to identify molecular markers linked to genes influencing this variation. PMID- 11870057 TI - Placental growth hormone and lactogen production by perifused ovine placental explants: regulation by growth hormone-releasing hormone and glucose. AB - The factors controlling normal placental development are poorly understood. We have previously reported the presence of ovine placental growth hormone (oPGH) and growth hormone receptors in ovine placenta, and oPGH production by the trophectoderm and syncitium during the second month of pregnancy. To identify factors regulating oPGH production, we developed a perifusion system to measure oPGH and ovine placental lactogen (oPL) production by Day 45 ovine placental explants. The mRNAs for both hormones were quantitated by real-time polymerase chain reaction in explants collected after perifusion periods of up to 8 h. Ovine PGH and oPL were released into the medium at mean rates of 2.45 +/- 0.2 and 353.6 +/- 13.6 ng/g/h, respectively. Ovine placenta produces growth hormone-releasing hormone (GHRH), but addition of GHRH to the perifusion medium did not modify either oPGH or oPL production. In vivo, oPGH production occurs between Days 30 and 60 of pregnancy. Because modulation of the maternal diet during this period affects placental development, the potential regulation of oPGH and oPL production by glucose was evaluated. Glucose supplementation of the perifusion medium resulted in a concentration-dependent decrease in oPGH release after 4 h, but oPGH mRNA levels were not affected. Production of oPL was not affected by glucose. Thus, oPGH and oPL belong to the same growth hormone/prolactin family but are differentially regulated by glucose. Ovine PGH modulations should be taken into account in metabolic experiments performed during the first trimester of pregnancy in sheep. PMID- 11870058 TI - Promoting effect of beta-mercaptoethanol on in vitro development under oxidative stress and cystine uptake of bovine embryos. AB - The effects of beta-mercaptoethanol (beta-ME) on in vitro development under oxidative stress and cystine uptake of bovine embryos were investigated. Bovine 1 cell embryos obtained by in vitro fertilization were cultured in TCM-199 or synthetic oviductal fluid (SOF) in 20% O(2) supplemented with beta-ME. Addition of beta-ME significantly (P < 0.01) promoted embryo development when cultured in both TCM-199 and SOF under high levels of O(2), to almost the same rates when they were cultured in 5% O(2). To investigate whether the growth-promoting effect of beta-ME was related to cystine uptake, which is an important amino acid for intracellular glutathione (GSH) synthesis, 1-cell, 8-cell, morula, and blastocyst stage embryos were incubated in cystine, cysteine-free TCM-199 containing radioisotope-labeled cystine supplemented with or without beta-ME. It was found that cystine uptake was consistently low in each embryo stage incubated without beta-ME. In contrast, addition of beta-ME significantly (P < 0.05 to 0.0001) promoted cystine uptake in each stage of embryo development. This increase of cystine uptake by beta-ME was significantly inhibited by supplementation of buthionine sulfoximine, a specific inhibitor of GSH biosynthesis (P < 0.0001). High-performance liquid chromatography (HPLC) analysis clearly revealed a decrease of cystine in culture medium after supplementation by beta-ME, thereby forming another peak. HPLC analysis also showed the incorporated cystine by supplementation of beta-ME was possibly metabolized for GSH synthesis in the embryos. These results indicate that beta-ME has a protective effect in embryo development against oxidative stress and that the effect of beta-ME is associated with the promotion of cystine uptake of low availability in embryos. PMID- 11870059 TI - Connexin43 in rat oocytes: developmental modulation of its phosphorylation. AB - It is well established that the 43-kDa connexin (Cx43) is predominantly expressed by ovarian somatic cells, whereas the identity of the connexins contributed by the oocyte to form gap junctions with its neighboring cells is not fully elucidated. Our study aimed to examine oocytes for the expression and regulation of Cx43 throughout oogenesis. Growing and fully grown rat oocytes that were meiotically incompetent and competent, respectively, were examined. Fully grown oocytes were analyzed either before or after reinitiation of meiosis as well as at the second meiotic metaphase. Immunofluorescent analysis of zona pellucida free oocytes using conventional and confocal microscopy demonstrated a characteristic pattern of punctuated staining of Cx43 on the oolema. Immunogold electron microscopy localized Cx43 to the oocyte surface and the microvillar processes. Reverse transcriptase-polymerase chain reaction and Western blot analysis revealed similar amounts of Cx43 gene and protein in oocytes of different developmental stages. However, a relative increase in the phosphorylated forms of the protein was observed in fully grown oocytes that had completed their maturation. Our findings demonstrate that rat oocytes express a developmentally regulated Cx43. They further suggest that homotypic gap junctions that consist of Cx43 may be present between rat oocytes and their adjacent cumulus cells. PMID- 11870060 TI - Detection of bradykinin and bradykinin-beta(2) receptors in the porcine endometrium during the estrous cycle and early pregnancy. AB - During the period of attachment of the trophectoderm to the uterine lumenal surface in the pig, there is an increase in uterine blood flow and a localized hyperemic response induced by the developing conceptuses. The presence of tissue kallikrein in the porcine uterine lumen suggests that the kallikrein-kinin system may be functional during pregnancy in the pig. The objective of the present study was to determine the concentration of bradykinin within the uterine lumen during the estrous cycle and early pregnancy as well as endometrial gene expression and cellular localization of the bradykinin beta(2) receptor. Concentration of bradykinin in uterine flushings was greatest during estrus (Day 0) and Days 12-18 of the estrous cycle. However, there was a 5- to 10-fold increase in bradykinin content in pregnant uterine flushings on Days 12-18 of pregnancy compared with the estrous cycle. Endometrial bradykinin beta(2) receptor gene expression was greatest on Days 0, 12, 15, and 18 of the estrous cycle and pregnancy as gene expression decreased almost 6-fold on Days 5 and 10. Bradykinin beta(2) receptors were detected in the endometrial surface and glandular epithelium with greatest intensity of staining observed on Days 0, 12, 15, and 18 of the estrous cycle and pregnancy. Results from the present study suggest that the kallikrein-kinin system plays a role in the establishment of pregnancy in the pig. PMID- 11870061 TI - Regulation of mitogen-activated protein kinase phosphorylation, microtubule organization, chromatin behavior, and cell cycle progression by protein phosphatases during pig oocyte maturation and fertilization in vitro. AB - We used okadaic acid (OA), a potent inhibitor of protein phosphatases 1 and 2A, to study the regulatory effects of protein phosphatases on mitogen-activated protein (MAP) kinase phosphorylation, morphological changes in the nucleus, and microtubule assembly during pig oocyte maturation and fertilization in vitro. When germinal vesicle (GV) stage oocytes were exposed to OA, MAP kinase phosphorylation was greatly accelerated, being fully activated at 10 min. However, MAP kinase was dephosphorylated by long-term (>20 h) exposure to OA. Correspondingly, premature chromosome condensation and GV breakdown were accelerated, whereas meiotic spindle assembly and meiotic progression beyond metaphase I stage were inhibited. OA also quickly reversed the inhibitory effects of butyrolactone I, a specific inhibitor of maturation-promoting factor (MPF), on MAP kinase phosphorylation and meiosis resumption. Treatment of metaphase II oocytes triggered metaphase II spindle elongation and disassembly as well as chromosome alignment disruption. OA treatment of fertilized eggs resulted in prompt phosphorylation of MAP kinase, disassembly of microtubules around the pronuclear area, chromatin condensation, and pronuclear membrane breakdown, but inhibited further cleavage. Our results suggest that inhibition of protein phosphatases promptly phosphorylates MAP kinase, induces premature chromosome condensation and meiosis resumption as well as pronucleus breakdown, but inhibits spindle organization and suppresses microtubule assembly by sperm centrosomes in pig oocytes and fertilized eggs. PMID- 11870062 TI - Analysis of differential messenger RNA expression between bovine blastocysts produced in different culture systems: implications for blastocyst quality. AB - Using reverse transcriptase-amplified fragment length polymorphism (RT-AFLP) analysis of differential mRNA expression and semiquantitative reverse transcriptase-polymerase chain reaction, we compared mRNA expression in bovine blastocysts from 4 sources, known to differ in quality in terms of their ability to withstand cryopreservation: 1) in vitro culture in synthetic oviduct fluid of in vitro-matured (IVM)/in vitro fertilized (IVF) zygotes; 2) in vitro culture in TCM-199 supplemented with granulosa cells (coculture) of IVM/IVF zygotes; 3) in vivo culture in the ewe oviduct of IVM/IVF zygotes; or 4) superovulation, artificial insemination, and nonsurgical embryo recovery. Total mRNA was isolated from pools of blastocysts and reverse transcription was performed. Triplicate reactions from each sample were displayed, and only consistent banding variations were recorded. Using AFLP-differential display assay, we found that cDNA banding patterns are highly conserved between the 4 groups of blastocysts studied; however, there was a difference of 7% in bands either missing or expressed across the groups. Fifty bands were reamplified, and a sequence comparison search revealed similarity of 14 isolated fragments to ribosomal and mitochondrial genes, 16 matched to described cDNA, and 20 corresponded to unknown sequences that may represent novel genes. The study of 7 differentially expressed mRNAs known to be involved in developmental process in the embryo suggests roles for apoptosis, oxidative stress, gap junctions, and differentiation in the determination of embryo quality. The aberrant transcription patterns detected in in vitro-produced bovine embryos compared with those produced in vivo may explain their reduced quality in terms of viability after cryopreservation. PMID- 11870064 TI - Estradiol regulation of luteinizing hormone secretion in heifers of two breed types that reach puberty at different ages. AB - The working hypothesis was that 17 beta-estradiol (E(2)) negative feedback on the hypothalamic-pituitary axis in regulation of LH secretion decreases during peripuberty in heifers of 2 different genotypes. We investigated whether Bos indicus heifers had a period postpuberty, as compared with prepuberty, of greater E(2) inhibition of LH secretion at a time when heifers of this genotype have been reported to have a period of anestrus. Prepubertal heifers 9 mo of age of 2 genotypes (B. indicus and B. taurus) were assigned to 3 groups (6 animals/group) to either remain intact (control), be ovariectomized, or be ovariectomized and implanted with E(2). Variables evaluated from 10 to 28 mo of age were circulating concentrations of progesterone (P(4)), presence of corpora lutea, and pulsatile pattern of LH release. Results confirmed that B. taurus heifers attained puberty at younger ages (P < 0.001) and at lower live weights (P = 0.015) than did B. indicus heifers (507 +/- 37 days of age vs. 678 +/- 7 days of age; 259 +/- 14 kg vs. 312 +/- 11 kg; respectively). There was cessation of E(2) inhibition of LH pulses coincident with the onset of puberty in heifers of both breed types but at a much younger age in B. taurus heifers. There was no evidence of enhanced negative feedback of E(2) on LH secretion subsequent to puberty in B. indicus heifers nor was there cessation of estrous cycles in control heifers of either breed type after puberty. PMID- 11870063 TI - Estrogen increases CD38 gene expression and leads to differential regulation of adenosine diphosphate (ADP)-ribosyl cyclase and cyclic ADP-ribose hydrolase activities in rat myometrium. AB - Hormones influence uterine contractility through their effects on intracellular calcium. The regulation of intracellular calcium in uterine smooth muscle is achieved by several mechanisms and includes mobilization from intracellular stores by inositol 1,4,5-trisphosphate and ryanodine-sensitive channels. Cyclic ADP-ribose (cADPR), a metabolite of NAD(+), is known to mediate calcium release through ryanodine receptor channels. A cell surface glycoprotein, CD38, catalyzes the synthesis and breakdown of cADPR and thus possesses bifunctional enzymatic activity. The regulation of cADPR synthesis by ADP-ribosyl cyclase (cyclase) or degradation by cADP-ribose hydrolase (hydrolase) by hormones in the myometrium is poorly understood. We investigated the effects of estradiol-17 beta on CD38 expression and the synthesis and degradation of cADPR in myometrial smooth muscle obtained from ovariectomized rats. CD38 expression was studied by reverse transcription polymerase chain reaction and Western blot analyses. In uterine microsomal fractions, cyclase and hydrolase activities were measured using nicotinamide guanine dinucleotide and [(32)P]cADPR as substrates, respectively. Microsomal proteins subfractionated by SDS-PAGE and gel filtration were used to determine the fractions containing cyclase and hydrolase activities. The results demonstrate that cyclase and hydrolase activities are associated with a single protein fraction, similar to CD38 in uteri from both ovariectomized and estradiol treated rats, and estradiol-17 beta causes 1) increased CD38 mRNA and protein expression and 2) significantly enhanced cyclase but not hydrolase activity. The differential regulation of CD38 by estradiol-17 beta, resulting in increased cADPR synthesis, would have profound effects on calcium regulation and myometrial contractility. PMID- 11870065 TI - Transient masculinization in the fossa, Cryptoprocta ferox (Carnivora, Viverridae). AB - In at least 9 mammalian species, females are masculinized throughout life, but the benefits of this remain unclear despite decades of thorough study, in particular of the spotted hyaena (Crocuta crocuta) in which the phenomenon has been associated with a high fitness cost. Through examination of wild and captive fossas (Cryptoprocta ferox, Viverridae), androgen assays, and DNA typing for confirmation of gender, we made the first discovery of transient masculinization of a female mammal. Juvenile female fossas exhibited an enlarged, spinescent clitoris supported by an os clitoridis and a pigmented secretion on the underpart fur that in adults was confined to males. These features appeared to diminish with age. The majority of adult females lacked them, and os clitoridis length was inversely related to head-body length. No evidence was found to link this masculinization to elevated female androgen levels. Circulating concentrations of testosterone and androstenedione, but not dihydrotestosterone, were significantly lower in females than in males. No significant differences in testosterone, androstenedione, or dihydrotestosterone levels were found between juvenile (masculinized) and adult (nonmasculinized) females. There are several possible physiological mechanisms for this masculinization. None of the hypotheses so far proposed to explain the evolutionary basis of female masculinization in mammals are applicable to our findings. We present 2 new hypotheses for testing and development. PMID- 11870066 TI - Cloning and characterization of porcine ovarian estrogen receptor beta isoforms. AB - The cDNA for the full-length porcine estrogen receptor beta (ER beta) and an alternatively spliced transcript with a deletion of exon 5 (ER beta delta 5) was cloned from pig ovary. RNase protection assays revealed that ER beta mRNA was expressed in the preovulatory follicles and early, midluteal, and regressing corpora lutea (CL) of eCG +/- hCG-primed gilts. ER beta and ER beta delta 5 transcripts were shown by semiquantitative reverse transcription polymerase chain reaction to be expressed at a ratio of approximately 2:1 in granulosa cells, small, medium, and large antral follicles, and midluteal phase corpora lutea of unprimed animals. Immunoreactive ER beta proteins corresponding to the size of in vitro translated ER beta and ER beta delta 5 were detected by immunoblot. Full length ER beta was detected in granulosa, small, medium, and large antral follicles, and midluteal phase CL of unprimed animals. Putative ER beta delta 5 immunoreactive bands were abundant only in granulosa cell extracts. In COS-1 cells, transfected ER beta delta 5 had no effect on basal transcription of an estrogen-responsive reporter construct but did repress wild-type ER beta transactivation when cotransfected at 10-fold excess plasmid. No repression of ER alpha transactivation was observed. In primary granulosa cell cultures, transfected ER beta delta 5 plasmid did not inhibit basal reporter activation. ER beta delta 5 was shown by immunofluorescence to localize to the nucleus in transfected COS-1 cells. In vitro translated ER beta delta 5 proteins bound estrogen response elements in DNA in electrophoretic mobility shift assays, as indicated by supershift analysis. ER beta is abundant in porcine ovary, and a naturally occurring splice variant missing exon 5 may have biological function. PMID- 11870067 TI - Monoclonal antibodies, immunofluorometric assay, and detection of human semenogelin in male reproductive tract: no association with in vitro fertilizing capacity of sperm. AB - Semenogelin plays an important role in sperm clotting and is degraded into smaller fragments by prostate-specific antigen (PSA) during clot liquefaction. Semenogelin and its fragments inhibit sperm motility in vitro. We studied the expression of semenogelin I mRNA and its localization in various tissues of the male genital tract. We also studied semenogelin concentrations with respect to sperm parameters and the outcome of in vitro fertilization. Semenogelin protein was detected by immunohistochemical staining and semenogelin I mRNA was detected by Northern blot analysis in the seminal vesicles and ampullary part of the vas deferens, whereas specimens from the prostate, epididymis, testis, and the female genital tract were negative. Using monoclonal antibodies against semenogelin, an immunofluorometric assay was developed to measure semenogelin levels in seminal plasma and to evaluate possible correlations with sperm parameters and fertilization in vitro. No correlation was found between the semenogelin concentration and the volume of the ejaculate, sperm concentration, sperm motility, or in vitro fertilization rate. Semenogelin levels were positively correlated with the total protein concentration in seminal plasma, and there was an inverse correlation between the concentration of semenogelin and that of PSA. The levels of semenogelin appear to bear no relationship to the in vitro fertilization capacity of the spermatozoa. PMID- 11870068 TI - Activation of the ribosomal RNA genes late in the third cell cycle of porcine embryos. AB - In porcine embryos, nucleoli are first observed during the third postfertilization cell cycle, i.e., at the 4-cell stage. However, direct studies of the initiation of rRNA transcription have not been reported. This transcription was investigated in the present study by simultaneous visualization of the rRNA genes and the rRNA by fluorescent in situ hybridization using a porcine 28S rDNA probe and subsequent visualization of argyrophilic nucleolar proteins by silver staining of extracted and fixed nuclei from in vivo-derived porcine embryos (n = 229). Nucleologenesis was observed by transmission electron microscopy. In general, the 2-cell and 4-cell embryos fixed at 10 and 20 h postcleavage (hpc) showed no signs of rRNA transcription. Four small clusters of fluorescein isothiocyanate (FITC) labeling were visible in interphase nuclei, consistent with hybridization to the rRNA gene clusters only; there was no silver staining at the sites of the rRNA genes and nucleolus precursor bodies. From 30 hpc onwards, most 4-cell embryos had medium size to large clusters of FITC labeled areas colocalized with silver staining of rRNA gene clusters and fibrillogranular nucleoli. These observations indicate that rRNA transcription had been initiated. These signs of rRNA synthesis could be blocked by actinomycin D, which is a strong inhibitor of RNA polymerase I. The rRNA transcription of porcine embryos is initiated between 20 and 30 hpc, corresponding to the end of the S-phase or the beginning of the G2 phase during the third cell cycle. PMID- 11870069 TI - Improvement of an electrical activation protocol for porcine oocytes. AB - Factors influencing pig oocyte activation by electrical stimulation were evaluated by their effect on the development of parthenogenetic embryos to the blastocyst stage to establish an effective activation protocol for pig nuclear transfer. This evaluation included 1) a comparison of the effect of epidermal growth factor and amino acids in maturation medium, 2) an investigation of interactions among oocyte age, applied voltage field strength, electrical pulse number, and pulse duration, and 3) a karyotype analysis of the parthenogenetic blastocysts yielded by an optimized protocol based on an in vitro system of oocyte maturation and embryo culture. In the first study, addition of amino acids in maturation medium was beneficial for the developmental competence of activated oocytes. In the second study, the developmental response of activated oocytes was dependent on interactions between oocyte age at activation and applied voltage field strength, voltage field strength and pulse number, and pulse number and duration. The formation of parthenogenetic blastocysts was optimal when activation was at 44 h of maturation using three 80-microsec consecutive pulses of 1.0 kV/cm DC. Approximately 84% of parthenogenetic blastocysts yielded by this protocol were diploid, implying a potential for further in vivo development. PMID- 11870071 TI - Local changes in blood flow within the early and midcycle corpus luteum after prostaglandin F(2 alpha) injection in the cow. AB - One of the postulated main luteolytic actions of prostaglandin (PG) F(2 alpha) is to decrease ovarian blood flow. However, before Day 5 of the normal cycle, the corpus luteum (CL) is refractory to the luteolytic action of PGF(2 alpha). Therefore, we aimed to determine in detail the real-time changes in intraluteal blood flow after PGF(2 alpha) injection at the early and middle stages of the estrous cycle in the cow. Normally cycling cows at Day 4 (early CL, n = 5) or Days 10--12 (mid CL, n = 5) of the estrous cycle (estrus = Day 0) were examined by transrectal color and pulsed Doppler ultrasonography to determine the blood flow area, the time-averaged maximum velocity (TAMXV), and the volume of the CL after an i.m. injection of a PGF(2 alpha) analogue. Ultrasonographic examinations were carried out just before PG injection (0 h) and then at 0.5, 1, 2, 4, 8, 12, 24, and 48 h after the injection. Blood samples were collected at each of these times for progesterone (P) determination. The ratio of the colored area to a sectional plane at the maximum diameter of the CL was used as a quantitative index of the changes in blood flow within the luteal tissue. Blood flow within the midcycle CL initially increased (P < 0.05) at 0.5-2 h, decreased at 4 h to the same levels observed at 0 h, and then further decreased to a lower level from 8 h (P < 0.05) to 48 h (P < 0.001). Plasma P concentrations decreased (P < 0.05) from 4.7 +/- 0.5 ng/ml (0 h) to 0.6 +/- 0.2 ng/ml (24 h). The TAMXV and CL volume decreased at 8 h (P < 0.05) and further decreased (P < 0.001) from 12 to 24 h after PG injection, indicating structural luteolysis. These changes were not detected in the early CL, in which luteolysis did not occur. In the early CL, the blood flow gradually increased in parallel with the CL volume, plasma P concentration, and TAMXV from Day 4 to Day 6. The present results indicate that PGF(2 alpha) induces an acute blood flow increase followed by a decrease in the midcycle CL but not in the early CL. This transitory increase may trigger the luteolytic cascade. The lack of intraluteal vascular response to PG injection in the early CL appears to be directly correlated with the ability to be resistant to PG. PMID- 11870070 TI - Somatic cell nuclear transfer in the pig: control of pronuclear formation and integration with improved methods for activation and maintenance of pregnancy. AB - To clone a pig from somatic cells, we first validated an electrical activation method for use on ovulated oocytes. We then evaluated delayed versus simultaneous activation (DA vs. SA) strategies, the use of 2 nuclear donor cells, and the use of cytoskeletal inhibitors during nuclear transfer. Using enucleated ovulated oocytes as cytoplasts for fetal fibroblast nuclei and transferring cloned embryos into a recipient within 2 h of activation, a 2-h delay between electrical fusion and activation yielded blastocysts more reliably and with a higher nuclear count than did SA. Comparable rates of development using DA were obtained following culture of embryos cloned from ovulated or in vitro-matured cytoplasts and fibroblast or cumulus nuclei. Treatment of cloned embryos with cytochalasin B (CB) postfusion and for 6 h after DA had no impact on blastocyst development as compared with CB treatment postfusion only. Inclusion of a microtubule inhibitor such as nocodozole with CB before and after DA improved nuclear retention and favored the formation of single pronuclei in experiments using a membrane dye to reliably monitor fusion. However, no improvement in blastocyst development was observed. Using fetal fibroblasts as nuclear donor cells, a live cloned piglet was produced in a pregnancy that was maintained by cotransfer of parthenogenetic embryos. PMID- 11870072 TI - Regulation of cyclic adenosine 3',5'-monophosphate response element binding protein (CREB) expression by Sp1 in the mammalian testis. AB - In the mammalian testis, the binding of FSH to Sertoli cells activates the cAMP dependent protein kinase A signaling pathway, resulting in the phosphorylation of the cAMP response element binding protein (CREB). Previous studies have also shown that CREB gene expression is activated by cAMP in Sertoli cells and that 2 cAMP response elements (CREs) that bind CREB and a neighboring Sp1 binding site are required for basal and cAMP-inducible CREB promoter activity. In contrast, CREB expression has been less well characterized in testis germ cells. We demonstrated that CREB and Sp1 are expressed in early germ cells only through the midpachytene stage of spermatogenesis. Furthermore, CREB promoter activity was induced over 70-fold by transient overexpression of Sp1 in SL2 cells, suggesting that Sp1 is an important regulator of CREB expression. Further studies of the CREB promoter revealed an additional regulatory element in the -130 region between the Sp1 and CREB transcription factor binding sites that is necessary for full promoter activity. Proteins expressed in Sertoli cells and germ cells bind specifically to the newly identified regulatory region. These studies suggest that proteins binding to Sp1 motifs and the -130 region are required to activate the CREB promoter. PMID- 11870073 TI - Capacitative calcium entry mechanism in porcine oocytes. AB - The presence of the capacitative Ca(2+) entry mechanism was investigated in porcine oocytes. In vitro-matured oocytes were treated with thapsigargin in Ca(2+)-free medium for 3 h to deplete intracellular calcium stores. After restoring extracellular calcium, a large calcium influx was measured by using the calcium indicator dye fura-2, indicating capacitative Ca(2+) entry. A similar divalent cation influx could also be detected with the Mn(2+)-quench technique after inositol 1,4,5-triphosphate-induced Ca(2+) release. In both cases, lanthanum, the Ca(2+) permeable channel inhibitor, completely blocked the influx caused by store depletion. Heterologous expression of Drosophila trp in porcine oocytes enhanced the thapsigargin-induced Ca(2+) influx. Polymerase chain reaction cloning using primers that were designed based on mouse and human trp sequences revealed that porcine oocytes contain a trp homologue. As in other cell types, the capacitative Ca(2+) entry mechanism might help in refilling the intracellular stores after the release of Ca(2+) from the stores. Further investigation is needed to determine whether the trp channel serves as the capacitative Ca(2+) entry pathway in porcine oocytes or is simply activated by the endogenous capacitative Ca(2+) entry mechanism and thus contributes to Ca(2+) influx. PMID- 11870076 TI - Expression patterns of retinoid X receptors, retinaldehyde dehydrogenase, and peroxisome proliferator activated receptor gamma in bovine preattachment embryos. AB - In cattle, administration of retinol at the time of superovulation has been indirectly associated with enhanced developmental potential of the embryo. Vitamin A and its metabolites influence several developmental processes by interacting with 2 different types of nuclear receptors, retinoic acid receptors and retinoid X receptors (RXRs). Given the limited information available concerning the RXR-mediated retinoid signaling system, particularly in species other than rodents, this study was performed to gain insight into the potential role of retinoid signaling during preattachment embryo development in the cow. Bovine embryos were produced in vitro from oocytes harvested from abattoir ovaries and frozen in liquid nitrogen at the oocyte, 2-, 4-, 8-, and 16- to 20 cell, morula, blastocyst, and hatched blastocyst stages. Reverse transcription polymerase chain reaction (PCR) and whole mount in situ hybridization were utilized to investigate mRNA expression for RXR alpha, RXR beta, RXR gamma, alcohol dehydrogenase I (ADH-I), retinaldehyde dehydrogenase 2 (RALDH2), peroxisome proliferator activated receptor gamma (PPAR gamma), and glyceraldehyde 3-phosphate dehydrogenase. Transcripts for RXR alpha, RXR beta, RALDH2, and PPAR gamma were detected in all stages beginning from the oocyte through to the hatched blastocyst. Whole mount in situ hybridization performed using digoxigenin labeled antisense probes detected all 4 transcripts in both the inner cell mass and the trophectoderm of hatched blastocysts. PCR products obtained for ADH-I exhibited very low homology to known human and mouse sequences. Immunohistochemistry was performed using polyclonal anti-rabbit antibodies against RXR beta and PPAR gamma to investigate whether these embryonic mRNAs were translated to the mature protein. Strong immunostaining was observed for both RXR beta and PPAR gamma in the trophectoderm and inner cell mass cells of intact and hatched blastocysts. Messenger RNA was not detected at any stage for RXR gamma. Expression of mRNA for RXR alpha, RXR beta, RALDH2, and PPAR gamma suggests that the early embryo may be competent to synthesize retinoic acid and regulate gene expression during preattachment development in vitro. PMID- 11870074 TI - Sp1 and Egr1 regulate transcription of the Dmrt1 gene in Sertoli cells. AB - Dmrt1 is a recently described gene that is specifically expressed in the gonads and is required for postnatal testis differentiation. Here, we describe the transcriptional mechanisms regulating the Dmrt1 proximal promoter in testicular Sertoli cells. A genomic clone containing exon 1 of the rat Dmrt1 gene and more than 9 kilobases of 5' flanking sequence was isolated and characterized. Several prominent transcriptional start sites were identified, with the major site located 102 bases from the translational start. The Dmrt1 5' flanking region from -5000 to +74 was transcriptionally active in primary Sertoli cells, and deletion analysis of this fragment identified 2 major regions needed for full Dmrt1 promoter function. These regions were located between -3200 and -2000 base pairs (bp) and downstream of -150 bp relative to the major transcriptional start site. DNase I footprint analysis of the region downstream of -150 bp revealed 3 regions that are bound by proteins from Sertoli cell nuclear extracts. Site-directed mutagenesis of these regions identified 2 elements that activate the Dmrt1 promoter and 2 that repress it. The positive elements bind the transcription factors Sp1, Sp3, and Egr1, suggesting that these transcription factors play a critical role in Dmrt1 regulation in the testis. PMID- 11870075 TI - Matrix metalloproteinase expression and activity following prostaglandin F(2 alpha)-induced luteolysis. AB - Luteal tissue contains matrix metalloproteinases (MMPs) that cleave specific components of the extracellular matrix (ECM) and are inhibited by tissue inhibitors of metalloproteinases (TIMPs). We previously reported a decrease in luteal TIMP-1 within 15 min of prostaglandin F(2 alpha) (PGF(2 alpha))-induced luteolysis. An increase in the MMP:TIMP ratio may promote ECM degradation and apoptosis, as observed in other tissues that undergo involution. The objectives of these experiments were to determine whether 1) PGF(2 alpha) affects expression of mRNA encoding fibrillar collagenases (MMP-1 and -13), gelatinases A and B (MMP 2 and -9), membrane type (mt)-1 MMP (MMP-14), stromelysin (MMP-3), and matrilysin (MMP-7), and 2) PGF(2 alpha) increases MMP activity during PGF(2 alpha)-induced luteolysis in sheep. Corpora lutea (n = 3-10/time point) were collected at 0, 15, and 30 min and 1, 2, 4, 6, 12, 24, and 48 h after PGF(2 alpha) administration. Northern blot analysis confirmed the presence of all MMPs except MMP-9. Expression of mRNA for the above MMPs (except MMP-2) increased significantly (P < 0.05) by 30 min, and all MMPs increased significantly (P < 0.05) by 6 h after PGF(2 alpha) administration. Expression of MMP-14 mRNA increased significantly (P < 0.05) by 15 min post-PGF(2 alpha) and remained elevated through 48 h. MMP activity in luteal homogenates (following proenzyme activation and inactivation of inhibitors) was increased significantly (P < 0.05) by 15 min and remained elevated through 48 h post-PGF(2 alpha). MMP activity was localized (in situ zymography) to the pericellular area of various cell types in the 0-h group and was markedly increased by 30 min post-PGF(2 alpha). MMP mRNA expression and activity were significantly increased following PGF(2 alpha) treatment. Increased MMP activity may promote ECM degradation during luteolysis. PMID- 11870077 TI - Thyroid hormones mediate steroid-independent seasonal changes in luteinizing hormone pulsatility in the ewe. AB - Thyroid hormones permit the increase in response to estradiol negative feedback in ewes at the transition to anestrus. In this study, we tested whether the thyroid hormones are also required for steroid-independent seasonal changes in pulsatile LH secretion. In experiment 1, Suffolk ewes were ovariectomized and thyroidectomized (THX) or ovariectomized only (controls) in late November. LH pulse frequency and amplitude were measured for 4 h in December, April, May, June, and August. Pulse frequency was also measured in the presence of estradiol containing implants during the breeding (December) and early anestrus (March) seasons. As expected, in the presence of estradiol, pulse frequency declined between December and March in control but not THX ewes. In the absence of estradiol, a seasonal decline in frequency and an increase in amplitude occurred in control ewes, concurrent with lengthening photoperiod. A similar trend was seen in THX ewes, but the seasonal changes were lower in magnitude and not significant. In experiment 2, the same protocol was used (pulse measurements in December, May, and June) with a larger THX group size (n = 7). Results were similar to those of experiment 1 for controls. In THX ewes, pulse frequency did not change over time and was significantly elevated relative to that of controls during the summer. Pulse amplitude in THX ewes tended to increase during summer and did not differ from pulse amplitudes in control ewes. These results demonstrate that thyroid hormones are required for steroid-independent cycles in LH pulse frequency; however, some seasonal changes in amplitude still occur in the absence of thyroid hormones. This finding contrasts with the changes in estradiol negative feedback at the transition to anestrus, which are entirely thyroid hormone dependent. PMID- 11870078 TI - Expression of hyaluronan synthases and CD44 messenger RNAs in porcine cumulus oocyte complexes during in vitro maturation. AB - The transient synthesis and accumulation of hyaluronan (HA), an extracellular matrix component of cumulus cells, brings about expansion of cumulus-oocyte complexes (COCs) in preovulatory mammalian follicles. In this study, we investigated the mRNA expressions of hyaluronan synthase 2 (has2), hyaluronan synthase 3 (has3), and CD44, as well as the responsiveness to eCG and porcine follicular fluid (pFF) of these genes, in porcine COCs, oocytectomized complexes (OXCs), and oocytes during in vitro maturation. Immunolocalization of CD44 was also analyzed in COCs. After 12 h of culture, the area of cumulus expansion in medium 199 supplemented with both 10 IU/ml eCG and 10% (v/v) pFF was significantly greater than that in the medium supplemented with eCG or pFF. Oocytectomy reduced the expansion area in the group supplemented with eCG. In reverse transcription-polymerase chain reaction analysis, all transcripts were identified in COCs, but has3 transcript was not found in OXCs. Only has3 mRNA was detectable in oocytes, indicating that cumulus cells express has2 and CD44 mRNAs, and oocytes express has3 mRNA. The expression levels of has2 and CD44 mRNAs in COCs and OXCs increased in the presence of eCG and pFF after 24 h of culture, suggesting that these genes have a positive dependency on eCG and pFF. In contrast, the high level of has3 mRNA was detected in COCs cultured in the medium alone. Oocytectomy slightly reduced the expression level of has2 mRNA. On immunostaining for CD44, CD44 was expressed apparently in COCs cultured with eCG and pFF for 24 h. The positive staining was distributed on cytoplasm along the perimembrane of cumulus cells and at the junctions between cumulus cells and oocytes. CD44 was also localized on cytoplasm of some oocytes. These results indicate that 1) porcine oocytes promote eCG-dependent cumulus expansion and the expression of has2 mRNA in cumulus cells, but these are not essential for expansion of cumulus cells and the expression of has2 mRNA; 2) HAS2 is involved in HA synthesis during cumulus expansion, and eCG and pFF up-regulate its expression; 3) the expression profile of the has3 mRNA that is transcribed in oocytes is different from those of has2 and CD44 mRNA; and 4) CD44 may participate in the interaction between cumulus cells and oocytes. PMID- 11870079 TI - Analysis of osteopontin at the maternal-placental interface in pigs. AB - Noninvasive, epitheliochorial placentation in the pig follows a prolonged preimplantation period characterized by migration, spacing and elongation of conceptuses, and secretion of estrogen for maternal recognition of pregnancy. Osteopontin (OPN) is an extracellular matrix protein that binds integrins to promote cell-cell attachment and communication. OPN appears to play a key role in conceptus implantation and maintenance of pregnancy in sheep; however, a role for OPN in the porcine uterus has not been established. Therefore, this study examined OPN expression and function in the porcine uterus and conceptus (embryo/fetus and associated extraembryonic membranes). Northern and slot blot hybridization detected an increase in endometrial OPN expression between Days 25 and 30, and levels remained elevated through Day 85 of pregnancy. In situ hybridization localized OPN mRNA to discrete regions of the uterine luminal epithelium (LE) on Day 15 of pregnancy and to the entire LE thereafter. Glandular epithelial (GE) expression of OPN mRNA was first detected on Day 35 of pregnancy and increased through Day 85. Both 70- and 45-kDa forms of OPN protein were detected in cyclic and pregnant endometrium by Western blotting. OPN protein was localized to the LE and GE by immunofluorescence; however, only the 70-kDa OPN was detected in uterine flushings. OPN protein was present along the entire uterine-placental interface after Day 30 of pregnancy. In addition, OPN mRNA and protein were localized to immune-like cells within the stratum compactum of the endometrium in both Day 9 cyclic and pregnant gilts. Incubation of OPN-coated microbeads with porcine trophectoderm and uterine luminal epithelial cells induced Arg-Gly-Asp (RGD)-dependent integrin activation and transmembrane accumulation of cytoskeletal molecules at the apical cell surface as assessed by immunofluorescence detection of talin or alpha-actinin as markers for focal adhesions. These results suggest that OPN, expressed by uterine epithelium and immune cells, may interact with receptors (i.e., integrins) on conceptus and uterus to promote conceptus development and signaling between these tissues as key contributors to attachment and placentation in the pig. PMID- 11870080 TI - Transgene insertion induced dominant male sterility and rescue of male fertility using round spermatid injection. AB - Transgene insertions in the mouse often cause mutations at chromosomal loci. Analysis of insertion mutations that cause male sterility may lead to the identification of novel molecular mechanisms implicated in male fertility. Here we show a line of transgenic mice with dominant inheritance of male sterility (DMS) that was found amid several lines that were normally fertile. Transgene positive males from this line invariably were sterile, whereas transgenic females and transgene-negative male littermates were fertile. Histologic analysis and TUNEL staining for apoptotic cells in DMS testis showed spermatogenesis arrest at metaphase of meiosis I (M-I), accompanied by massive apoptosis of spermatocytes. Meiosis I arrest was incomplete, however, as small numbers of spermatids and spermatozoa were found. Both round spermatids and spermatozoa were evaluated for their permissiveness in the assisted reproductive technologies intracytoplasmic sperm injection (ICSI) and round spermatid injection (ROSI). Surprisingly, ROSI but not ICSI gave live offspring, suggesting that mature sperm had deteriorated by the time of recovery from the epididymis. Mapping the transgene insertion by fluorescence in situ hybridization revealed a site on chromosome 14 D3-E1. Two candidate genes, GFR alpha 2 and GnRH, that were previously mapped to that region and the functions of which in spermatogenesis are well established were not altered in DMS. As a consequence, positional cloning of the DMS locus will be essential to identify new molecules potentially involved in arrest at M-I. Furthermore, mice carrying this genetic trait might be useful for studies of assisted reproductive technologies and male contraceptives. PMID- 11870082 TI - Inhibition of platelet-derived growth factor actions in the embryonic testis influences normal cord development and morphology. AB - Platelet-derived growth factors (PDGFs) are paracrine factors with roles in mesenchymal-epithelial interactions during normal and pathologic processes. Previously, PDGF and its receptor (PDGFR) have been shown to be present in perinatal, peripubertal, and adult rat testes. The role of PDGF in embryonic testicular cord formation is not known. The hypothesis tested is that PDGFs and PDGFRs are expressed during cord formation and that inhibition of their action influences normal cord formation during embryonic testis development. Embryonic Day (E) 13 gonadal organ cultures were used. Organs were cultured for 3 days and treated daily with vehicle or a PDGFR-specific tyrosine phosphorylation inhibitor (i.e., the tyrphostin AG1295 or AG1296). Vehicle-treated testes formed normal cords, whereas tyrphostin-treated testes formed "swollen cords," a phenomenon characterized by a significant decrease in the number of cords per testis area and increased cord diameter due to fusion of cords. Expression of PDGF and PDGFR in E13, E14, E16, Postnatal Day (P) 0, and P20 testes was examined. Messenger RNAs for PDGF-A and -B and PDGF alpha- and beta-receptors were expressed in isolated testes during all developmental periods examined. Immunoreactivity for PDGF was present throughout the testicular compartment at E14, restricted primarily to testicular cords at E16, and present in cells of the testicular cords with a stronger immunoreactivity in certain interstitial cell types of P0 testis. PDGFR beta-receptor immunoreactivity was primarily localized to the mesonephros of E14 organs and the testicular interstitium of E16 and P0 testes. Tyrphostins did not affect apoptotic cell number in the testis. PDGF had no effect on cell growth in P0 testis cultures. The results show that PDGFs and PDGFRs are expressed in embryonic testis during cord formation in a tissue specific manner. Inhibition of PDGF actions does not inhibit cord formation but does alter normal cord development and morphology. The observations provide insight into the factors involved in male sex differentiation and embryonic testis development. PMID- 11870081 TI - SAMP32, a testis-specific, isoantigenic sperm acrosomal membrane-associated protein. AB - To identify novel human sperm membrane antigens, we analyzed two-dimensional gels of sperm extracts containing hydrophobic proteins that partitioned into Triton X 114. Four protein spots with isoelectric points (pIs) ranging from 4.5 to 5.5 and apparent molecular weights from 32 to 34 kDa were sequenced by mass spectrometry and found to contain common peptide sequences. Cloning the corresponding cDNA revealed that these protein spots were products of a single gene (SAMP32), encoding a protein of 32 kDa with a predicted pI of 4.57. SAMP32 has a potential transmembrane domain in the carboxyl terminus and is phosphorylated in vivo on serine 256. Northern blotting of eight human tissues and RNA dot blotting of 76 human tissues showed that SAMP32 expression was testis specific. SAMP32 contained an amino terminal domain homologous to the major malarial circumsporozoite surface protein and a domain similar to that of Krp1 from Schizosaccharomyces pombe in its carboxyl terminus. The SAMP32 locus consists of seven exons on chromosome 6q15-16.2. Antiserum against recombinant SAMP32 recognized protein spots originally cored from a two-dimensional gel. This antiserum strongly stained the equatorial segment and faintly stained the acrosome cap of ejaculated human spermatozoa by immunofluorescence. Immunoelectron microscopy showed that SAMP32 was associated with the inner acrosomal membrane in the principal and the equatorial segments of the sperm acrosome. By immunostaining enzyme-dissociated testicular cells, SAMP32 was localized to Golgi phase round spermatids and subsequent stages of acrosome biogenesis. Recombinant SAMP32 reacted with serum from an infertile man, suggesting that it is isoantigenic. Antibodies against recombinant SAMP32 inhibited both the binding and the fusion of human sperm to zona-free hamster eggs. PMID- 11870083 TI - Fas-Fas ligand system mediates luteal cell death in bovine corpus luteum. AB - Fas antigen (Fas) is a cell surface receptor that triggers apoptosis in sensitive cells when bound to the Fas ligand (Fas L). The present study was undertaken to identify the presence of a Fas-Fas L system in bovine corpus luteum (CL) and to evaluate the regulation of Fas-mediated luteal cell death by leukocyte-derived cytokines. The reverse transcription-polymerase chain reaction showed higher levels of Fas mRNA expression in CL in the regressed luteal stage (Days 19-21) than in the other stages (P < 0.05). Bovine luteal cells from midcycle CL (Days 8 12) were exposed for 24 h to interferon gamma (IFN; 50 ng/ml) and/or tumor necrosis factor alpha (TNF; 50 ng/ml). After 24 h of culture, the expression of Fas mRNA was detected in the cultured cells and was increased by IFN. Moreover, TNF augmented the stimulatory action of IFN, whereas TNF alone did not affect the expression of Fas mRNA. The effects of IFN and TNF on Fas-mediated cell death were also examined. Cells were exposed to IFN and/or TNF for 24 h and were further treated with IFN and/or TNF in the presence or absence of Fas L (100 ng/ml) for 24 h. Treatments of the cells with IFN alone and in combination with TNF resulted in killing of 30% and 50% of the cells (P < 0.05), respectively, whereas TNF alone did not have a cytotoxic effect on the cells. On the other hand, Fas L killed 60% of the cells treated with IFN (P < 0.01) and 85% of the cells treated with the combination of TNF and IFN (P < 0.01), respectively, whereas Fas L showed no effect on the viability of the luteal cells treated with or without TNF. Furthermore, shrunken nuclei and apoptotic bodies were observed in the cells treated with Fas L in the presence of TNF and IFN. The overall results suggest that a Fas-Fas L system is present in bovine CL and that leukocyte-derived TNF and IFN play important roles in Fas-mediated luteal cell death. PMID- 11870084 TI - Molecular cloning of porcine estrogen receptor-beta complementary DNAs and developmental expression in periimplantation embryos. AB - In the pig, estrogens transiently produced by embryos and progestins of maternal origin target the uterine endometrium, causing alterations in gene expression and secretory activity, both of which are important for the initiation of embryo attachment. The potential direct embryotrophic roles of estrogens and progestins are, however, unknown. Here we report the cloning of porcine embryonic estrogen receptor-beta (ER-beta) mRNA by reverse transcription-polymerase chain reaction (RT-PCR) using specific primer sets designed initially within conserved regions of human and bovine ER-beta mRNAs, and subsequently within regions of identified porcine ER-beta cDNA sequences. The ER-beta mRNA has an open reading frame of 1578 nucleotides and encodes a 526 amino acid polypeptide that displays greater than 90% identity with other mammalian ER-beta proteins. Northern and Western blot analyses using porcine filamentous embryos from Day 12 of pregnancy demonstrated the presence of multiple ER-beta mRNA transcripts of approximately 9.5, 4.9, and 3.5 kilobases, and a similar 64-kDa protein corresponding in size to human ovarian granulosa cell ER-beta, respectively. In Day 12 filamentous embryos, ER-beta expression was immunolocalized to trophoblastic cell nuclei, coincident with that of proliferative cell nuclear antigen (PCNA). The developmental ontogeny of ER-beta mRNA was evaluated in embryos of different morphologies (spherical, tubular, and filamentous) by semiquantitative RT-PCR, along with those for other steroid hormone receptors (ER-alpha and progesterone receptor) and known embryonic genes associated with cell differentiation (cytochrome P450 aromatase type III) and growth (cyclin D1). ER-beta mRNA levels varied with embryo morphology (filamentous maximum at Day 12), coincident with that of cyclin D1. Progesterone receptor mRNA levels were maximal in tubular embryos, similar to that of P450 aromatase, whereas the expression of the ER alpha gene was barely detectable and appeared constitutive for all developmental stages examined. Estradiol-17 beta treatment of Day 12 filamentous embryos in culture up-regulated ER-beta and P450 aromatase (type III) mRNA levels, respectively, but decreased those of PCNA, and had no effect on cyclin D1 mRNA levels. These studies taken together suggest that embryonic ER-beta likely mediates the autocrine functions of estrogens in the dynamic regulation of embryonic growth and development at periimplantation. PMID- 11870085 TI - Clinostat rotation induces apoptosis in luteal cells of the pregnant rat. AB - Recent studies have shown that microgravity induces changes at the cellular level, including apoptosis. However, it is unknown whether microgravity affects luteal cell function. This study was performed to assess whether microgravity conditions generated by clinostat rotation induce apoptosis and affect steroidogenesis by luteal cells. Luteal cells isolated from the corpora lutea of Day 8 pregnant rats were placed in equal numbers in slide flasks (chamber slides). One slide flask was placed in the clinostat and the other served as a stationary control. At 48 h in the clinostat, whereas the levels of progesterone and total cellular protein decreased, the number of shrunken cells increased. To determine whether apoptosis occurred in shrunken cells, Comet and TUNEL assays were performed. At 48 h, the percentage of apoptotic cells in the clinostat increased compared with that in the control. To investigate how the microgravity conditions induce apoptosis, the active mitochondria in luteal cells were detected with JC-1 dye. Cells in the control consisted of many active mitochondria, which were evenly distributed throughout the cell. In contrast, cells in the clinostat displayed fewer active mitochondria, which were distributed either to the outer edge of the cell or around the nucleus. These results suggest that mitochondrial dysfunction induced by clinostat rotation could lead to apoptosis in luteal cells and suppression of progesterone production. PMID- 11870087 TI - Molecular cloning and characterization of a complementary DNA encoding sperm tail protein SHIPPO 1. AB - Formation of the tail in developing sperm is a complex process involving the organization of the axoneme, transport of periaxonemal proteins from the cytoplasm to the tail, and assembly of the outer dense fibers and fibrous sheath. Although detailed morphological descriptions of these events are available, the molecular mechanisms remain to be fully elucidated. We have isolated a new gene, named shippo 1, from a haploid germ cell-specific cDNA library of mouse testis, and also its human orthologue (h-shippo 1). The isolated cDNA is 1.2 kilobases long, carrying a 762-base pair open reading frame that encodes SHIPPO 1, a sperm protein predicted to consist of 254 amino acids. The amino acid sequence includes 6 Pro-Gly-Pro repeats, which are also present in the human orthologue protein (hSHIPPO 1) as well as in 2 other newly reported proteins of Drosophila melanogaster. Transcription of shippo 1 is exclusively observed in haploid germ cells. Antibody raised against SHIPPO 1 identified a testis-specific M(r) 32 x 10(-3) band in Western blot analysis. The protein was further localized in the flagella of the elongated spermatids and along the entire length of the tail in mature sperm. SHIPPO 1 in sperm is resistant to treatment with nonionic detergents and coextracted with the cytoskeletal core proteins of the mouse sperm tail. PMID- 11870086 TI - Relaxin depresses small bowel motility through a nitric oxide-mediated mechanism. Studies in mice. AB - Gastrointestinal motility is reduced and the incidence of functional gastrointestinal disorders is increased in pregnancy, possibly due to hormonal influences. This study aims to clarify whether the hormone relaxin, which attains high circulating levels during pregnancy and has a nitric oxide-mediated relaxant action on vascular and uterine smooth muscle, also reduces bowel motility and, if it does, whether nitric oxide is involved. Female mice in proestrous or estrous were treated for 18 h with relaxin (1 microg s.c.) or vehicle (controls). Isolated ileal preparations from both groups were used to record contractile activity, either basal or after acute administration of relaxin (5 x 10(-8) M). Drugs inhibiting nitric oxide biosynthesis or neurotransmission were used in combination with relaxin. Expression of nitric oxide synthase isoforms by the ileum was assessed by immunocytochemistry and Western blot analysis. Relaxin caused a clear-cut decay of muscle tension and a reduction in amplitude of spontaneous contractions upon either chronic administration to mice or acute addition to isolated ileal preparations. These effects were significantly blunted by N(G)-nitro-L-arginine, but not by the neural blockers we used. Moreover, relaxin increased the expression of nitric oxide synthases II and III, but not synthase I. Relaxin markedly inhibits ileal motility in mice by exerting a direct action on smooth muscle through the activation of intrinsic nitric oxide biosynthesis. PMID- 11870088 TI - Preservation of the wild European mouflon: the first example of genetic management using a complete program of reproductive biotechnologies. AB - Although the potential use of reproductive biotechnologies for safeguarding endangered wildlife species is undoubted, practical efforts have met with limited success to date. In those instances in which modern technologies have been adapted to rescuing rare or endangered species, procedures have been applied piecemeal, and no consistent breeding program based on reproductive biotechnologies has been undertaken. Here we describe for the first time the rescue of an endangered species, the European mouflon (Ovis orientalis musimon), by the application of an integrated package of reproductive biotechnologies. This genetic management extended from the initial collection of gametes, through the in vitro production of embryos and interspecific transfer, to the birth of healthy mouflon offspring. In addition, a genetic resource bank for the European mouflon was established, with cryopreserved sperm, embryos, and somatic cells. PMID- 11870089 TI - Hemochorial placentation in the primate: expression of vascular endothelial growth factor, angiopoietins, and their receptors throughout pregnancy. AB - Vascular development and its transformation are necessary for successful hemochorial placentation, and vascular endothelial growth factor (VEGF), angiopoietins, and their receptors may be involved in the molecular regulation of this process. To determine the potential role of these putative regulators in a widely studied primate, the common marmoset, we investigated their mRNA expression and protein location in the placenta throughout pregnancy using in situ hybridization, Northern blot analysis, and immunocytochemistry. VEGF was localized in decidual and cytotrophoblast cells, and its highest expression was found in the maternal decidua. The Flt receptor was exclusively detected in the syncytial trophoblast with increasing expression in placentae from 10 wk to term. Soluble Flt (sFlt) was also detectable by Northern blot analysis. KDR receptor expression was restricted to mesenchymal cells during early placentation and to the fetoplacental vasculature during later placentation. KDR expression increased throughout pregnancy. Angiopoietin-1 (Ang-1) was localized in the syncytial trophoblast, being highly expressed in the second half of gestation. Ang-2 mRNA localized exclusively to maternal endothelial cells, and was highly expressed in 10-wk placentae. The Tie-2 receptor was found in cytotrophoblast cells and in fetal and maternal vessels. High Tie-2 levels were detected in the wall of chorion vessels at 14-wk, 17-wk, and term placentae. These results suggest that the processes of trophoblast invasion, maternal vascular transformation, and fetoplacental vascular differentiation and development are regulated by the specific actions of angiogenic ligand-receptor pairs. Specifically, 1) VEGF/Flt and Ang-1/Tie-2 may promote trophoblast growth, 2) VEGF/KDR and Ang-1/Tie-2 may support fetoplacental vascular development and stabilization, 3) sFlt may balance VEGF actions, and 4) Ang-2/Tie-2 may remodel the maternal vasculature. PMID- 11870090 TI - Involvement of protein kinase C and arachidonic acid pathways in the gonadotropin releasing hormone regulation of oocyte meiosis and follicular steroidogenesis in the goldfish ovary. AB - The involvement of protein kinase C (PKC) and arachidonic acid (AA) pathways were investigated in the GnRH regulation of oocyte meiosis and follicular testosterone production in the goldfish ovary. The results clearly demonstrate differences in the postreceptor mechanisms involving the stimulatory and inhibitory actions of GnRH peptides on basal and gonadotropin (GtH)-induced reinitiation of oocyte meiosis and steroidogenesis. In isolated goldfish follicles in vitro, the observed stimulatory effects of both salmon GnRH (sGnRH) and chicken GnRH-II (cGnRH-II) on germinal vesicle breakdown were completely blocked by addition of PKC inhibitors, suggesting the involvement of PKC, presumably through activation of phospholipase C/diacylglycerol pathways in the GnRH-induced reinitiation of oocyte meiosis. Administration of an AA metabolism inhibitor, however, only blocked the stimulatory effect of sGnRH without affecting cGnRH-II-induced meiosis. As observed previously, in the presence of GtH, sGnRH was found to inhibit GtH-induced resumption of meiosis and testosterone production, whereas cGnRH-II was without effect. The inhibitory effect of sGnRH on GtH-induced meiosis and steroidogenesis was completely reversed by addition an AA metabolism inhibitor, whereas PKC inhibitors had no effect. These findings provide functional evidence in support of the novel hypothesis that goldfish ovarian follicles contain GnRH-receptor subtypes with different ligand selectivity mediating stimulatory and inhibitory actions of sGnRH and cGnRH in the goldfish ovary. PMID- 11870091 TI - Surface expression of complement receptor gC1q-R/p33 is increased on the plasma membrane of human spermatozoa after capacitation. AB - Evidence is increasing that complement components might play a role in fertilization. C1q, the first component of the classical complement cascade, has the ability to promote sperm agglutination in a capacitation-dependent manner as well as an effect on sperm-oolemma binding and fusion. We have previously detected gC1qR, the receptor for the globular head portion of C1q, on the surface of capacitated sperm. In this study, we examined the expression of gC1qR in both fresh and capacitated human spermatozoa. We performed immunoprecipitation for gC1qR and analyzed biotinylated sperm membrane by Western blot to illustrate an increase in receptor density after overnight capacitation. These results were confirmed by flow cytometric analysis of spermatozoa using fluorescein isothiocyanate-labeled monoclonal anti-gC1qR antibody. Confocal, indirect immunofluorescence microscopy revealed an increase in receptor expression over the rostral portion of the sperm head after capacitation. In addition, the ability of live spermatozoa to bind to monoclonal anti-gC1qR antibody-coated microtiter wells was also increased after capacitation. These results suggest that gC1qR may play a role in human fertilization. PMID- 11870092 TI - Pituitary adenylate cyclase-activating polypeptide modulates plasminogen activator expression in rat granulosa cell. AB - Pituitary adenylate cyclase-activating polypeptide (PACAP) is a bioactive peptide isolated from ovine hypothalamus. It has been demonstrated to be transiently expressed in preovulatory follicles and to positively affect several parameters correlated with the ovulatory process. The aim of the present study was to investigate whether PACAP influences the plasminogen/plasmin system in rat ovary. Plasminogen activators (PAs) are serine proteases, modulated by gonadotropins and several peptides in preovulatory follicles, that appear to be involved in ovulation. Granulosa cells obtained from immature eCG-treated rats were cultured for 24 h in the presence of increasing concentrations of PACAP and vasoactive intestinal peptide (VIP). A significant, dose-dependent increase in tissue-type PA (tPA) activity and decrease in urokinase-type (uPA) PA activity were observed in PACAP-treated cells. These effects were exerted at the mRNA level. The use of cycloheximide, a protein synthesis inhibitor, suggested that PACAP requires an intermediary protein to decrease uPA-mRNA, but not to induce tPA-mRNA. However, no significant modulation of PAs was observed in the presence of VIP. When granulosa cells were stimulated within the intact follicle (i.e., maintaining the three-dimensional structure and in the presence of the theca cell layers), both PACAP and VIP dose-dependently stimulated tPA. These data suggest that, in addition to the PACAP type I receptor present on granulosa cells, different subtypes of PACAP receptors are present in the different ovarian compartments. PMID- 11870093 TI - Developmental potential in bovine oocytes is related to cumulus-oocyte complex grade, calcium current activity, and calcium stores. AB - A morphological classification of the immature cumulus-oocyte complex (COC), which grossly resembled the atresia grade of its follicle source, was used in bovine oocytes to determine 1) the developmental potential by either in vitro fertilization or parthenogenetic activation, 2) the calcium current activity by whole-cell voltage clamp technique, and 3) the intracytoplasmic calcium stores by microfluorimetric evaluation. The COC classification took into account some cumulus and ooplasm features, designated as follows: A) presence of a clear and compact cumulus and translucent ooplasm, B) dark and compact cumulus and dark ooplasm, and C) dark and expanded cumulus and dark ooplasm. We found no difference between in vitro fertilization and parthenogenetically activated oocytes in terms of cleavage rate and blastocyst production. Both protocols indicated a significant variability between the three compared COC categories. The B-COCs showed the highest embryo production efficiency as well as the greatest Ca(2+) current activity, whereas A-COCs showed an opposite pattern. The C-COCs, mostly attributed to atretic and heavily atretic follicles, showed morphological characteristics between those of A- and B-COCs. Stores of Ca(2+) were significantly greater in A-COCs than in B- and C-COCs in the case of immature oocytes, and greater in B-COCs than in C-and A-COCs in the case of in vitro-matured oocytes. These results demonstrate that in the bovine 1) the considered morphological criteria for oocyte classification are related to developmental competence, 2) plasma membrane Ca(2+) current in the immature oocyte is related to developmental potential, and 3) calcium stores are related to morphological quality in immature oocytes and to developmental competence in mature oocytes. PMID- 11870096 TI - Chlamydia trachomatis in subfertile women undergoing uterine instrumentation. Screen or treat? AB - Women who undergo uterine instrumentation are considered at risk for pelvic infections by Chlamydia trachomatis, which may derive either from ascending endocervical infections or from reactivation of micro-organisms persisting in the genital tract after previous chlamydia infections. Women presenting at fertility clinics frequently undergo uterine instrumentation (e.g. hysterosalpingography and laparoscopy with hydrotubation). Studies in subfertile women using DNA amplification techniques have shown that the prevalence of endocervical chlamydia infections is low (1.8%). In contrast, in 30-60% of subfertile women chlamydia IgG antibodies can be found in serum, indicating previous chlamydia infections. It has been demonstrated that, several years after chlamydia infections, viable micro-organisms may still be present in the upper genital tract. Therefore, subfertile women with chlamydia antibodies should be considered at risk for reactivation of persistent chlamydia infections after uterine instrumentation, even after exclusion of endocervical chlamydia infections. Moreover, in subfertile women without chlamydia antibodies, the presence of viable micro organisms in the genital tract cannot be excluded. As a consequence, prophylactic antibiotics before uterine instrumentation should be considered in all subfertile women, instead of endocervical screening for C. trachomatis and treatment of positive cases only. PMID- 11870094 TI - KRP3A and KRP3B: candidate motors in spermatid maturation in the seminiferous epithelium. AB - We have identified KRP3, a novel kinesin-related protein expressed in the mammalian testis, and have examined the tissue distribution and subcellular localization of isoforms of this protein. Isolation of KRP3 clones, using the head domain identified in a previous PCR screen as probe, identified at least two KRP3 isoforms in the rat. We have isolated coding sequences of two highly related cDNAs from the rat testis that we have termed KRP3A and KRP3B (kinesin-related protein 3, A and B). Both cDNAs code for predicted polypeptides with the three domain structure typical of kinesin superfamily members; namely a conserved motor domain, a region capable of forming a limited coiled-coil secondary structure, and a globular tail domain. Although almost identical in their head and stalk domains, these motors diverge in their tail domains. This group of motors is found in many tissues and cell types. The KRP3B motor contains DNA-binding motifs and an RCC1 (regulator of chromosome condensation 1) consensus sequence in its tail domain. Despite this similarity, KRP3B is not associated with the same structures as RCC1. Instead, KRP3 isoforms localize with the nuclei of developing spermatids, and their immunolocalization in the testis overlaps with that of the small GTPase Ran. Like Ran, KRP3 motors are associated in a polarized fashion with the nucleus of maturing spermatids at various stages of elongation. Our findings suggest a possible role for KRP3 motor isoforms in spermatid maturation mediated by possible interaction with the Ran GTPase. PMID- 11870097 TI - Alternative treatments in reproductive medicine: much ado about nothing. "The fact that millions of people do not master arithmetic does not prove that two times two is anything else than four": W.F. Hermans. AB - The popularity of alternative medicine certainly also affects patients suffering from infertility. Alternative medicine started in the seventies but there have always been unorthodox practitioners, treating infertile women and men. Some historical examples will be described. The claims made for alternative medicine in the lay press have not been accompanied by similar reports in the mainstream medical journals. Practitioners and advocates of alternative medicine have used several strategies to defend their position. These were mostly of a philosophical nature, but more recently the practitioners of alternative medicine admit that the effectiveness of their therapies should be proven in randomized trials, as is considered mandatory in regular medicine. There are very few well-designed papers on the effectiveness of alternative medicine with the exception of one kind of paper that is hard for editors of medical journals to resist: seemingly impeccable papers proving absurd claims, whose mechanisms of action are, for instance, completely incomprehensible. We argue that this type of paper should be rejected for publication and indeed offer explanations for their mere existence. PMID- 11870098 TI - Ethical considerations on preimplantation genetic diagnosis for HLA typing to match a future child as a donor of haematopoietic stem cells to a sibling. AB - Recently, several requests were made by couples with an affected child who wanted preimplantation genetic diagnosis (PGD) to select embryos in the hope of conceiving an HLA identical donor sibling. This article considers the ethical arguments for and against the application of PGD for this goal. Only embryos HLA matched with an existing sibling in need of a compatible donor of haematopoietic stem cells would be transferred. The main arguments are the instrumentalization of the child, the best-interests standard, the postnatal test for acceptability and the experience of the donor child. It is argued that conceiving a child to save a child is a morally defensible decision on the condition that the operation that will be performed on the future child is acceptable to perform on an existing child. The instrumentalization of the donor child does not demonstrate disrespect for its autonomy or its intrinsic worth. PMID- 11870099 TI - The immunological "Wars of the Roses": disagreements amongst reproductive immunologists. AB - The relevance of abnormal autoimmune function to reproductive function in the female has over recent years become an increasingly controversial and contentious issue. Opposing views have led to a polarization of opinions which, at times, resulted in publications of rather vocal opinions by individuals as well as societal committees. This communication is an attempt to reconcile these, at times diametrically opposing opinions, in a concept of (auto)immune-driven reproduction failure, which could explain and unify these opposing opinions and, thus, hopefully end the ongoing "immunological wars of the roses". PMID- 11870100 TI - Laparoscopic salpingectomy for women with hydrosalpinges enhances the success of IVF: a Cochrane review. AB - BACKGROUND: The aim of this study was to determine whether surgical intervention is effective for women with tubal disease who are due to undergo treatment with IVF. METHODS: A systematic review employing the principles of the Cochrane Menstrual Disorders and Subfertility Group was undertaken. Three randomized controlled trials were included, the population of women in all three studies having hydrosalpinges. RESULTS: The odds of pregnancy [odds ratio (OR) = 1.75, 95% confidence interval (CI) 1.07-2.86] and of ongoing pregnancy and live birth (OR = 2.13, 95% CI 1.24-3.65) were increased with laparoscopic salpingectomy for hydrosalpinges prior to IVF. There were no significant differences in the odds of embryo implantation (OR = 1.34, 95% CI 0.87-2.05), ectopic pregnancy (OR = 0.42, 95% CI 0.08-2.14), miscarriage (OR = 0.49, 95% CI 0.16-1.52) or treatment complications (OR = 5.80, 95% CI 0.35-96.79). No data were available concerning the odds of multiple pregnancy or the proportion of IVF cycles resulting in embryo transfer. CONCLUSION: Laparoscopic salpingectomy should be considered for all women with hydrosalpinges due to undergo IVF; further research is required to assess other pre-IVF surgical interventions (such as needle aspiration of hydrosalpinx fluid, laparoscopic proximal tubal occlusion and laparoscopic salpingostomy) for women with hydrosalpinges. PMID- 11870101 TI - Immediate post-partum insertion of intrauterine devices: a Cochrane review. AB - BACKGROUND: Insertion of an intrauterine device (IUD) immediately after delivery is appealing for several reasons. The woman is known not to be pregnant, her motivation for contraception may be high and the setting may be convenient for both the woman and her provider. However, the risk of spontaneous expulsion may be unacceptably high. The objective of this study was to assess the efficacy and feasibility of IUD insertion immediately after expulsion of the placenta. Our a priori hypothesis was that this practice is safe, but associated with higher expulsion rates than interval IUD insertion. METHODS: We used Medline, Popline, Embase, and Cochrane Controlled Trials Register computer searches, supplemented by review articles and contact with investigators. We sought all randomized controlled trials that had at least one treatment arm that involved immediate post-partum (within 10 min of placental expulsion) insertion of an IUD. Comparisons could include different IUDs, different insertion techniques, immediate versus delayed post-partum insertion, or immediate versus interval insertion (unrelated to pregnancy). Studies could include either vaginal or Caesarean deliveries. We evaluated the methodological quality of each report and sought to identify duplicate reporting of data from multicentre trials. We abstracted data onto data collection forms. Principal outcome measures included pregnancy, expulsion and continuation rates. Because the trials did not have uniform interventions, we were unable to aggregate them in a meta-analysis. RESULTS: We found no randomized controlled trials that directly compared immediate post-partum insertion with either delayed post-partum or interval insertion. Modifications of existing devices, such as adding absorbable sutures or additional appendages, did not appear beneficial. Most studies showed no important differences between insertions done by hand or by instruments. Lippes Loops and Progestasert devices did not perform as well as copper devices. CONCLUSIONS: Immediate post-partum insertion of IUDs appeared safe and effective, though direct comparisons with other insertion times were lacking. Advantages of immediate post-partum insertion include high motivation, assurance that the woman is not pregnant, and convenience. However, expulsion rates appear to be higher than with interval insertion. The popularity of immediate post-partum IUD insertion in countries as diverse as China, Mexico and Egypt support the feasibility of this approach. Early follow-up may be important in identifying spontaneous IUD expulsions. PMID- 11870102 TI - Genetic factors contribute to the risk of developing endometriosis. AB - BACKGROUND: Endometriosis is known to cluster within nuclear families. The extent of familial clustering can be evaluated in Iceland with its large population based genealogical database. METHODS AND RESULTS: Applying several measures of familiality we demonstrated that 750 women with endometriosis were significantly more interrelated than matched control groups. The risk ratio for sisters was 5.20 (P < 0.001) and for cousins 1.56 (P = 0.003). The average kinship coefficient for the patients was significantly higher than that calculated for 1000 sets of 750 matched controls (P < 0.001) and this remained significant when contribution from first-degree relatives was excluded (P < 0.05). The minimum number of ancestors required to account for the group of patients was compared with the minimum number of ancestors required to account for the control groups at different time points in the past. The minimum number of founders for the group of patients was significantly smaller than for the control groups. Affected cousin pairs were as likely to be paternally connected as maternally connected. CONCLUSIONS: This is the first population-based study using an extensive genealogy database to examine the genetic contribution to endometriosis. A genetic factor is present, with a raised risk in close and more distant relatives, and a definite kinship factor with maternal and paternal inheritance contributing. PMID- 11870103 TI - Studies on the human leukocyte antigen-DR in patients with endometriosis: genotyping of HLA-DRB1 alleles. AB - BACKGROUND: An increasing number of reports suggest that endometriosis is associated with abnormal immune function involving changes in both cell-mediated and humoral immunity, although the aetiology of the disease remains undefined. The human leukocyte antigen system (HLA) is known to play a role in the aetiology of a number of diseases. This study examines the possible association between the HLA-DR and endometriosis. METHODS: Eighty-three patients diagnosed with endometriosis by laparoscopic examination were typed for HLA-DR antigens using a PCR-restriction fragment length polymorphism analysis. The frequency of HLA-DR genotypes in this patient population was compared with that in a population of general controls. RESULTS: The incidence of HLA-DRB1*1403 in the patient group was 6.0% (10/166 alleles) compared with 1.4% in the control group (6/444 alleles), and the incidence of the HLA-DRB1*1403 allele was significantly greater in patients with endometriosis compared with the control population [odds ratio 4.68, 95% confidence interval 1.67-13.09, P = 0.0013 (chi(2) analysis), (corrected P values: Pc = 0.0494)]. No significant difference was found between the two groups with regards to the frequency of the other HLA-DR alleles. CONCLUSIONS: The results from this study may be helpful to implicate a role of HLA-DR antigens in the development of endometriosis, although further investigation is required. PMID- 11870104 TI - Molecular and cytogenetic characterization of an azoospermic male with a de-novo Y;14 translocation and alternate centromere inactivation. AB - BACKGROUND: Y-autosome (Y/A) translocations have been reported in association with male infertility. Different hypotheses have been made as to correlations between Y/A translocations and spermatogenetic disturbances. We describe an azoospermic patient with a de-novo Y;14 translocation: 45,X,dic(Y;14)(q12;p11). METHODS AND RESULTS: Cytogenetic, fluorescent in-situ hybridization (FISH) and molecular studies have been performed. A 14/22 (D14Z1/D22Z1) centromere and a Y centromere (DYZ1) probe both showed a signal on the translocation chromosome, confirming its dicentricity. Each copy of the translocation chromosome had only one primary constriction, with inactivation of the Y centromere in most (90%) of the cells. The 14 centromere was inactive in the remaining cells (10%). FISH and molecular deletion mapping analysis allowed acute assignment of the Yq breakpoint to the junction of euchromatin and heterochromatin (Yq12), distal to the AZF gene location (Yq11). CONCLUSIONS: This study supports the hypothesis that in Y/A translocations infertility might be related to meiotic disturbances with spermatogenetic arrest. In addition, sex chromosome molecular investigations, performed on single spermatids, suggest a highly increased risk of producing chromosomally abnormal embryos. PMID- 11870105 TI - Chromosome analysis of epididymal and testicular sperm in azoospermic patients undergoing ICSI. AB - BACKGROUND: Although ICSI provides a way of treating azoospermic men, concern has been raised about the potential risk for transmission of genetic abnormalities to the offspring. We quantified the incidence of chromosomal abnormalities in epididymal and testicular sperm retrieved from azoospermic patients undergoing ICSI. METHODS: Individual testicular sperm were collected from testicular biopsies with an ICSI pipette, and epididymal sperm were retrieved by microsurgical epididymal sperm aspiration. Samples were processed by fluorescent in-situ hybridization (FISH) for chromosomes 18, 21, X and Y and the results compared with those from normal ejaculated samples. RESULTS: The overall aneuploidy rate of 11.4% in men with non-obstructive azoospermia was significantly higher (P = 0.0001) than the 1.8% detected in epididymal sperm from men with obstructive azoospermia and also the 1.5% found in ejaculated sperm. No significant difference was found between the epididymal and ejaculated samples. When the chromosomal abnormalities were analysed, gonosomal disomy was the most recurrent abnormality in both obstructive and non-obstructive azoospermic patients, while autosomal disomy was the most frequent in ejaculated sperm. CONCLUSIONS: Sperm of non-obstructive azoospermic men had a higher incidence of chromosomal abnormalities, of which sex chromosome aneuploidy was the most predominant. Genetic counselling should be offered to all couples considering infertility treatment by ICSI with testicular sperm. PMID- 11870106 TI - Sperm aneuploidy in fathers of children with paternally and maternally inherited Klinefelter syndrome. AB - BACKGROUND: It is unclear whether frequency of sperm aneuploidy is associated with risk of fathering children with trisomy. METHODS: We recruited 36 families with a boy with Klinefelter syndrome (KS), interviewed the fathers about their exposures and medical history, received a semen sample from each father, and collected blood samples from the mother, father and child. We applied a multicolour fluorescent in-situ hybridization assay to compare the frequencies of sperm carrying XY aneuploidy and disomies X, Y and 21 in fathers of maternally and paternally inherited KS cases. RESULTS: Inheritance of the extra X chromosome was paternal in 10 and maternal in 26 families. Fathers of paternal KS cases produced higher frequencies of XY sperm (P = 0.02) than fathers of maternal KS cases. After controlling for age, the major confounding variable, the difference between the two groups was no longer significant (P less-than-or-equal 0.2). Also, there were no significant differences between the parental origin groups for disomy X, Y or 21. CONCLUSIONS: Men who fathered a child with a Klinefelter syndrome produced higher frequencies of XY sperm aneuploidy, which is explained, in part, by both paternal age and parent of origin. PMID- 11870107 TI - Fallopian tube ciliary beat frequency in relation to the stage of menstrual cycle and anatomical site. AB - BACKGROUND: The cyclical changes in ciliary structure and motion within the human Fallopian tube are well documented. Previous investigators have studied ciliary beat frequency (CBF) in relation to menstrual cycle and anatomical site, but with conflicting results. METHODS: Using a technique that records variations in light intensity, we have studied the changes in CBF in relation to the menstrual cycle and anatomical site. Fallopian tubes were collected from 26 women who underwent hysterectomy for benign conditions. Menstrual history, hormone profile and endometrial biopsy results were used to determine the stage of the cycle. Fourteen women were in the proliferative phase, and 12 women in the secretory phase. RESULTS: Mean CBF for all subjects was 5.3 plus minus 0.2 Hz. There was no significant difference in CBF in relation to anatomical site. In the fimbrial region the ciliary beat was faster in the secretory (5.8 plus minus 0.3 Hz) as compared with the proliferative phase (4.9 plus minus 0.2 Hz), P < 0.02. CONCLUSIONS: It is possible that this increase in fimbrial CBF may contribute to ovum retrieval and transport after ovulation. However, the reproductive significance of the changes in CBF in relation to the menstrual cycle needs further investigation. PMID- 11870108 TI - Acute and chronic effects of genistein, tyrphostin and lavendustin A on steroid synthesis in luteinized human granulosa cells. AB - BACKGROUND: Phytoestrogens, including genistein and other inhibitors of tyrosine kinases (TKs), inhibit specific steroidogenic enzymes. This study was designed to compare the effects of genistein, with two other TK inhibitors, on steroid synthesis in human granulosa luteal (GL) cells and to identify which steroidogenic enzymes they may affect. METHODS: GL cells, obtained from women undergoing IVF procedures, were cultured for various periods of time with and without substrates for progesterone and estradiol synthesis, in the presence or absence of the TK inhibitors. RESULTS: The TK inhibitors significantly suppressed progesterone and estradiol synthesis in a dose-dependent manner over a 48 h culture period. Progesterone production in the presence of 10(-7) mol/l pregnenolone during a 4 h period was inhibited by both acute (4 h) and chronic (24 h) exposure of GL cells to 50 micromol/l genistein (P < 0.05) whilst no significant effects of 50 micromol/l tyrphostin A23 were observed. Genistein (4 and 24 h exposure) inhibited the production of estradiol using 10(-7) mol/l estrone as a substrate, but inhibition of estradiol synthesis using androstenedione or testosterone as substrates was only observed after a 24 h exposure. In contrast, tyrphostin acutely stimulated estradiol synthesis when androstenedione and testosterone were used as substrates (P < 0.05) but not estrone. CONCLUSIONS: Genistein directly inhibits 3 and 17beta-hydroxysteroid dehydrogenase activity, whilst tyrphostin has an acute stimulatory effect on aromatase activity. Over a longer time (24 and/or 48 h period), both TK inhibitors suppress steroid synthesis. PMID- 11870109 TI - Modulation of neutrophil apoptosis by plasma and peritoneal fluid from patients with advanced endometriosis. AB - BACKGROUND: The increased production of pro-inflammatory chemoattractant cytokines for neutrophils in endometriosis suggests that changes in the immune system play an important role in the pathophysiology of endometriosis. The effects of plasma and peritoneal fluid from patients with advanced endometriosis on the apoptosis of neutrophils were investigated. METHODS: Apoptotic changes of neutrophils were evaluated by morphological changes using Giemsa staining. Apoptosis was confirmed by DNA electrophoretic analysis. RESULTS: Compared with the plasma (n = 20) and peritoneal fluid (n = 5) of healthy controls, the addition of 10% plasma (n = 20) and peritoneal fluid (n = 10) from patients with endometriosis to an in-vitro culture of neutrophils from healthy subjects reduced the percentage of apoptotic cells from 65.3 +/- 6.6 to 27.2 +/- 4.6% (P < 0.001) and from 45.3 +/- 4.8 to 10.5 +/- 4.3% (P < 0.001) respectively. Neutralizing interleukin-8 antibody abrogated the delay of neutrophil apoptosis induced by peritoneal fluid, but not in the plasma of endometriosis patients. CONCLUSIONS: These findings show that interleukin-8 is one of the neutrophil survival factors in the peritoneal fluid of endometriosis patients and that an unidentified survival factor is also present in the plasma of patients with endometriosis. PMID- 11870110 TI - Relationship between oxidative stress and embryotoxicity of hydrosalpingeal fluid. AB - BACKGROUND: Oxidative stress mechanisms are involved in the pathophysiology of many reproductive disorders. The objective of this study was to characterize oxidative stress parameters in hydrosalpingeal fluid (HSF) and examine their possible role in early embryo development. METHODS AND RESULTS: HSF was aspirated at laparoscopic salpingectomy in 11 infertile women. Reactive oxygen species (ROS), total (non-enzymatic) antioxidant capacity (TAC) and lipid peroxidation (LPO) were assayed. Two-cell mouse embryos were incubated with 25, 50 or 75% HSF and the blastocyst development rate was observed. ROS was detected in five of 11 (45%) HSF samples with a mean of 4.2 x 10(4) c.p.m. LPO was detected in all samples at a mean (+/- SD) value of 5575.4 +/- 6091.9 micromol/l malonaldehyde. The mean blastocyst development rate at 25, 5+/- 0 and 75% HSF and in the control group was 88.9 +/- 9.4, 65.7 +/- 19.1, 45.7 +/- 5.7 and 96.7% respectively (P < 0.0001). The blastocyst development rate was positively correlated to ROS concentrations (P < 0.02) but was not significantly related to LPO. CONCLUSIONS: The blastocyst development rate decreased with increasing concentrations of HSF. For the first time, the presence of ROS, LPO and TAC activity in human HSF was characterized. A possible role of oxidative stress in the embryotoxicity of HSF is suggested. PMID- 11870111 TI - Early massive follicle loss and apoptosis in heterotopically grafted newborn mouse ovaries. AB - BACKGROUND: Ovarian tissue cryopreservation and transplantation can be used to restore fertility to sterile females. A question that warrants further investigation is whether the follicular content is affected by the freeze-thawing and grafting procedure, and if so, to what extent and by what mechanism. METHODS AND RESULTS: Intact newborn mouse ovaries were allografted under the kidney capsule or were cryopreserved by slow freezing with dimethylsulphoxide as the cryoprotectant prior to grafting. Estrogenic activity of ovariectomized recipient mice, as revealed by vaginal cytology, resumed after 11 days of transplantation. At 14 days after transplantation, ovarian grafts were recovered and processed histologically for follicle number counting. The follicular content of grafts of fresh ovaries was 58% of that from ovaries of age-matched 14 day old mice. In frozen-thawed ovarian grafts, the follicular content was only 9% lower than that of fresh grafted ovaries. Apoptosis of follicular cells was investigated by DNA nick end labelling. We observed a marked increase in the staining of fragmentation of DNA shortly after transplantation (2-12 h) of fresh newborn mouse ovaries. CONCLUSIONS: The results of the present study indicate that transplantation rather than cryopreservation accounts for the major and early loss of primordial follicles in grafted newborn mouse ovaries. PMID- 11870112 TI - Functional studies of subcutaneous ovarian transplants in non-human primates: steroidogenesis, endometrial development, ovulation, menstrual patterns and gamete morphology. AB - BACKGROUND: The study objective was to determine if ovarian function would be restored following fresh and cryopreserved extrapelvic autologous ovarian transplantation and if vascular endothelial growth factor (VEGF) administration would augment the success rate. METHODS: Sixteen regularly cycling female cynomolgus monkeys underwent bilateral oophorectomy and were randomly assigned to one of three treatment groups: (i) sham transplant group (n = 5) underwent transplantation of pieces of adipose tissue; (ii) fresh autologous ovarian transplantation without VEGF administration (n = 6) and (iii) fresh autologous ovarian transplantation with 1 microg of VEGF (n = 5) administered at the transplantation site daily for 14 days after transplantation. The ovarian tissue from the sham transplanted group was cryopreserved. This material was later thawed and transplanted in four out of five of the sham operated group. RESULTS: Five out of six (83%) of the primates in our transplantation group without VEGF had functioning ovarian transplants while two of five (40%) primates in our transplantation group with VEGF administration had functioning ovarian transplants. The cryopreserved group had two of four primates (50%) with functioning transplants. Ovarian stimulation yielded multiple follicles and one metaphase II oocyte from the fresh and one metaphase II oocyte from the cryopreserved group. CONCLUSIONS: The success of ovarian transplantation in these non-human primates with menstrual cycles bodes well for the development of ovarian transplantation protocols for women at risk of ovarian failure. PMID- 11870113 TI - Ovarian and adrenal steroid production: regulatory role of LH/HCG. AB - BACKGROUND: The contribution of the adrenal glands to the total circulating steroid pool in women is not well known. There is evidence that human adrenals express the LH receptor gene and that LH may affect adrenal androgen secretion. METHODS: HCG stimulation tests (a single dose of 5000 IU i.m.) were performed in women at reproductive age (group 1, n = 6, age 21--39 years) before and after treatment with a GnRH agonist for 3 weeks, and in oophorectomized post-menopausal women (group 2, n = 6, 47--59 years) during and after estrogen replacement therapy (ERT). RESULTS: HCG did not stimulate the secretion of cortisol, dehydroepiandrosterone (DHEA) and dehydroepiandrosterone sulphate (DHEAS) in group 2. In contrast, in group 1, the basal concentrations of serum 17 hydroxyprogesterone (17-OHP), androstenedione, testosterone and estradiol (E(2)) were stimulated significantly (17-OHP 105%, androstenedione 31%, testosterone 20%, E(2) 136%) by HCG, and the treatment with GnRH agonist decreased the responses. The basal serum concentrations of these steroids were significantly lower in oophorectomized women (17-OHP 57%, androstenedione 46%, testosterone 25%), and HCG did not increase these levels. It can be approximated that the ovarian contribution to the circulating levels of 17-OHP, androstenedione and testosterone is 25--30%, and that the adrenals are the primary source of cortisol, DHEA and DHEAS. CONCLUSION: LH/HCG does not have a major role in the regulation of adrenal steroid synthesis in endocrinologically healthy women. PMID- 11870114 TI - Predicting pregnancy and spermatogenesis by survival analysis during gonadotrophin treatment of gonadotrophin-deficient infertile men. AB - BACKGROUND: Predictors of fertility or spermatogenesis during gonadotrophin therapy of gonadotrophin-deficient men remain poorly defined. METHODS AND RESULTS: In order to evaluate potential predictors, this study evaluated 29 consecutive gonadotrophin-deficient men all desiring paternity who received 43 courses of therapy in one centre between 1982 and 1998. The Kaplan-Meier survival analysis estimates of median (SE) time to a sperm concentration of >0, >5 and >20 x 10(6)/ml were 5.5 (1.1), 12.4 (2.3) and 29.1 (1.9) months respectively. Conception occurred in 22/43 cycles (with eight men achieving two pregnancies) with a median (SE) Kaplan-Meier estimate of 20.5 (4.7) months. The median sperm concentration at conception was 5.0 (SE 2.0; range 0.0-59.5) x 10(6)/ml. Multivariate correlated Cox proportional hazards models predicting these same sperm thresholds and conception were developed by forward stepwise variable selection with verification of the model by backward stepping. Larger testicular volume, prior gonadotrophin therapy, completion of puberty, older age, the absence of adverse fertility factors and the absence of multiple pituitary hormone deficiency predicted a favourable response. Multivariate modelling suggests that the two most important predictors of sperm output are testicular volume and pubertal status. The most important potentially modifiable predictor was prior gonadotrophin therapy. The efficacy of recombinant and urinary FSH were similar. Prior androgen therapy and partner's age did not appear to be significant. CONCLUSIONS: Since prolonged treatment may be required to induce spermatogenesis, attention to these predictors may allow appropriate early use of advanced reproductive technologies. PMID- 11870115 TI - Women with poor response to IVF have lowered circulating gonadotrophin surge attenuating factor (GnSAF) bioactivity during spontaneous and stimulated cycles. AB - BACKGROUND: Up to 13% of IVF cancellations are due to poor responses during down regulated cycles. Because premature luteinization occurs more frequently in older or "poor responder" patients, defective production of gonadotrophin surge attenuating factor (GnSAF) may be involved. METHODS: Nine women with normal previous IVF response (NORM) and 9 with previous poor IVF response (POOR) were monitored in a spontaneous cycle (blood samples: days 2, 7, 11, 15 and 20) and then stimulated with recombinant human FSH (rFSH) under GnRH agonist (blood samples: treatment days GnRH agonist + 2, GnRH agonist + 7, day of HCG administration and days HCG + 1 and HCG + 8). LH, FSH, estradiol, progesterone and inhibin-A and -B were assayed in individual samples while GnSAF bioactivity was determined in samples pooled according to day, cycle and IVF response. RESULTS: During spontaneous cycles LH, steroids and inhibins were similar between NORM and POOR women, FSH was elevated in POOR women (4.9 +/- 0.3 versus 6.7 +/- 0.6 mIU/l, P < 0.01) and GnSAF bioactivity was detectable on days 2, 7 and 11 in NORM women only. During IVF cycles inhibin-A and -B rose more markedly in NORM than POOR women. Similarly GnSAF production peaked on day GnRH agonist + 7 in NORM women, but on the day of HCG administration in POOR women. CONCLUSIONS: Defects in ovarian responsiveness to FSH include reduced GnSAF production. This suggests that GnSAF should be investigated as a marker of ovarian reserve once an immunoassay becomes available. PMID- 11870116 TI - Follicle stimulating hormone measured in unextracted urine throughout the menstrual cycle correlates with age and ovarian reserve. AB - BACKGROUND: A method was previously described to measure FSH reliably in unextracted urine. The aim of the current study was to establish the course of FSH measured in urine throughout the cycle. METHOD: Daily urinary FSH (uFSH) concentrations were determined in 14 regularly menstruating volunteers aged 23-39 years during one complete menstrual cycle. RESULTS: In each subject, mean daily uFSH measured in urine, as gold standard for FSH tone, correlated significantly with FSH in early follicular phase fixed to menstruation on cycle day 3 (r = 0.75, P = 0.002), or fixed to ovulation 9 days before the pre-ovulatory FSH surge (r = 0.87, P = 0.0001), or when selected as being the highest follicular phase value (r = 0.91, P = 0.0001). Age correlated significantly with mean daily uFSH (r = 0.67, P = 0.009), highest follicular phase uFSH (r = 0.60, P = 0.024), uFSH on cycle day 3 (r = 0.80, P = 0.0006), and uFSH 9 days before FSH surge (r = 0.65, P = 0.0016). The uFSH was also measured on cycle day 3 in 104 IVF patients in a cycle prior to pituitary down-regulation. The uFSH correlated significantly with numbers of follicles (P = 0.02) and oocytes (P = 0.024). CONCLUSION: It is concluded that cycle day 3 uFSH is a good reflection of the mean uFSH of the complete cycle, and there is a highly significant correlation between uFSH and age and ovarian reserve. Measurement of FSH in urine on cycle day 3 seems to be a reliable and non-invasive tool for determining ovarian reserve in IVF. PMID- 11870117 TI - A pilot study of the long-term effects of acipimox in polycystic ovarian syndrome. AB - BACKGROUND: To evaluate the effects of long-term acipimox administration on glucose-induced insulin secretion and peripheral insulin sensitivity in polycystic ovarian syndrome (PCOS), 20 PCOS subjects (eight lean and 12 obese) and 14 body mass index-matched controls (seven lean and seven obese) were investigated. METHODS: Fasting blood samples were collected for basal hormone and lipoprotein assays, after which patients underwent an oral glucose tolerance test (OGTT). The following day a euglycaemic-hyperinsulinaemic clamp was performed. After 4-6 weeks of treatment with acipimox at a dose of 250 mg given orally three times a day, the patients repeated the study protocol. RESULTS: No significant differences were found in the glucose, insulin or C-peptide responses to OGTT before and after anti-lipolytic drug administration in any group, nor was there any effect on insulin sensitivity. Concerning the lipid profile, acipimox administration led to a significant decrease of cholesterol and low-density lipoprotein levels in obese PCOS patients as well as in obese and lean controls. Lower triglycerides were found after the drug administration in both obese groups. Post-treatment free fatty acid levels were not significantly different when compared with basal values. CONCLUSIONS: Acipimox does not appear to be an effective insulin-lowering drug in PCOS, even if it can be used in obese women with PCOS as an additional therapeutic agent to ameliorate the atherogenic lipid profile of the syndrome. PMID- 11870118 TI - Pilot study on the use of repeated doses of sublingual misoprostol in termination of pregnancy up to 12 weeks gestation: efficacy and acceptability. AB - BACKGROUND: A sublingual misoprostol-alone regimen was used in 50 women requesting medical abortion at up to 12 weeks gestation. The efficacy and acceptability of this regimen were studied. METHODS: The women were given 600 microg misoprostol sublingually every 3 h for a maximum of 5 doses. RESULTS: The overall complete abortion rate was 86% (95% confidence interval: 74-93). The mean number of doses of misoprostol required was 4.1 +/- 1.1. There was no significant change in haemoglobin concentration and the median duration of vaginal bleeding was 15 days (range: 7-56). Diarrhoea, fever and chills were the most common side effects. The acceptability of this regimen of misoprostol was good: 97.7% of the women who had a complete abortion would choose this method again and 88.4% would recommend it to others. They preferred sublingual misoprostol as it is convenient to take, avoids the painful vaginal administration and gives more privacy during the abortion process. CONCLUSION: This regimen of sublingual misoprostol is an effective and acceptable method of medical abortion. Randomized controlled trials are required to compare the efficacy of various misoprostol-alone regimens of medical abortion. Pharmacokinetic studies and clinical trials are needed to find out the most appropriate dose, dosing interval and route of administration of misoprostol. PMID- 11870119 TI - Adjuvant L-arginine treatment in controlled ovarian hyperstimulation: a double blind, randomized study. AB - BACKGROUND: Enhanced vascularization appears to be important for follicular selection and maturation in both spontaneous and stimulated IVF cycles. Nitric oxide, formed in vivo from L-arginine, may play a key role in follicular maturation and ovulation. METHODS: To evaluate the role of L-arginine supplementation in controlled ovarian hyperstimulation, 37 IVF patients were divided into two groups according to ovarian stimulation protocols: group I, GnRH agonist plus pure (p)FSH plus oral L-arginine (n = 18); and group II, GnRH agonist plus pFSH plus placebo (n = 19). Hormonal, ultrasonographic and Doppler evaluations were performed, and plasma and follicular fluid nitrite/nitrate concentrations were monitored. RESULTS: Thirty-two patients completed the study. In group I (n = 16), plasma L-arginine concentrations increased from (basal) 87 +/- 12 micromol to 279 +/- 31 micromol (P = 0.002) on the day of beta-HCG administration. In this group, pFSH treatment was shorter (P = 0.039) than in group II (n = 16). The number of the follicles > or =17mm was lower (P = 0.038) in group I than group II. The "good quality" embryos were fewer in number (P = 0.034) and pregnancy rate, both per patient (P = 0.024) and per embryo transfer (P = 0.019), was lower in group I. In the L-arginine group, an increased follicular fluid concentration of nitrite/nitrate was observed. On day 8 of the cycle, elevated plasma estradiol levels were associated with decreased blood flow resistances of perifollicular arteries. Follicular fluid concentrations of nitrite/nitrate were inversely correlated with embryo quality (r = -0.613; P = 0.005) and perifollicular artery pulsatility index (r = -0.609; P = 0.021). CONCLUSIONS: L-Arginine supplementation may be detrimental to embryo quality and pregnancy rate during controlled ovarian hyperstimulation cycles. PMID- 11870120 TI - The impact of the embryo transfer catheter on the pregnancy rate in IVF. AB - BACKGROUND: The aim was to assess whether the type of embryo transfer set used for embryo transfer affects the ongoing pregnancy rate in IVF. METHODS: The TDT set was compared with the K-soft 5000 in a large, prospective, randomized study. Patients were randomized moments before transfer by drawing a consecutively numbered, sealed, opaque envelope indicating the catheter to be used. RESULTS: 2059 embryo transfers in 1296 patients were analysed. The ongoing pregnancy rate was significantly higher in the K-soft group. If the first transfer of a patient (n = 1296) within this study period was analysed, the ongoing pregnancy rates were 27.1 versus 20.5% (P = 0.006). If the analysis is limited to patients that underwent their very first transfer ever (n = 607), the ongoing pregnancy rates were 30.3 versus 20.0% (P = 0.003) in favour of the K-soft. CONCLUSION: We conclude from these data that the type of embryo transfer set used for embryo transfer does affect the ongoing pregnancy rate and that the impact of the variable transfer catheter on the ongoing pregnancy rate increases when the a priori chance of pregnancy increases. PMID- 11870122 TI - The value of Chlamydia trachomatis antibody testing in predicting tubal factor infertility. AB - BACKGROUND: The objective of the present study was to compare the likelihood of abnormal Chlamydia trachomatis antibody test results with that of abnormal hysterosalpingography (HSG) test results in patients with tubal factor infertility. METHODS: Anti-C. trachomatis immunoglobulin G antibodies were determined prospectively in 295 infertility patients by means of an indirect fluorescent antibody technique. In 48 of the 295 patients both HSG and laparoscopy with chromotubation were performed. The results of C. trachomatis antibody testing were compared with the results of HSG with respect to their predictive value of tubal factor infertility. Likelihood ratios for abnormal C. trachomatis antibody and HSG test results were determined in infertility patients, as assessed by laparoscopy. RESULTS: The positive likelihood ratio for C. trachomatis antibody testing was 1.8. This was comparable with the HSG, which had a positive likelihood ratio of 1.7. CONCLUSIONS: The predictive value of C. trachomatis antibody testing was equal to that of HSG, but ratios of 1.7 and 1.8 indicate a poor test, so both C. trachomatis antibody testing and HSG have a poor predictive value. C. trachomatis antibody testing causes minimal inconvenience to the patient, in contrast to HSG, and therefore should be maintained in infertility examinations. PMID- 11870121 TI - Neonatal data on a cohort of 2889 infants born after ICSI (1991-1999) and of 2995 infants born after IVF (1983-1999). AB - BACKGROUND: To evaluate the safety of ICSI, this study compared data of IVF and ICSI children by collecting data on neonatal outcome and congenital malformations during pregnancy and at birth. METHODS: The follow-up study included agreement to genetic counselling and eventual prenatal diagnosis, followed by a physical examination of the children after 2 months, after 1 year and after 2 years. 2840 ICSI children (1991-1999) and 2955 IVF children (1983-1999) were liveborn after replacement of fresh embryos. ICSI was carried out using ejaculated, epididymal or testicular sperm. RESULTS: In the two cohorts, similar rates of multiple pregnancies were observed. ICSI and IVF maternal characteristics were comparable for medication taken during pregnancy, pregnancy duration and maternal educational level, whereas maternal age was higher in ICSI and a higher percentage of first pregnancies and first children born was observed in the ICSI mothers. Birthweight, number of neonatal complications, low birthweight, stillbirth rate and perinatal death rate were compared between the ICSI and the IVF groups and were similar for ICSI and IVF. Prematurity was slightly higher in the ICSI children (31.8%) than in the IVF children (29.3%). Very low birthweight was higher in the IVF pregnancies (5.7%) compared with ICSI pregnancies (4.4%). Major malformations (defined as those causing functional impairment or requiring surgical correction), were observed at birth in 3.4% of the ICSI liveborn children and in 3.8% of the IVF children (P = 0.538). Malformation rate in ICSI was not related to sperm origin or sperm quality. The number of stillbirths (born > or =20 weeks of pregnancy) was 1.69% in the ICSI group and 1.31% in the IVF group. Total malformation rate taking into account major malformations in stillborns, in terminations and in liveborns was 4.2% in ICSI and 4.6% in IVF (P = 0.482). CONCLUSIONS: The comparison of ICSI and IVF children taking part in an identical follow-up study did not show any increased risk of major malformations and neonatal complications in the ICSI group. PMID- 11870124 TI - The effects of cryopreservation on sperm morphology, motility and mitochondrial function. AB - BACKGROUND: The effects of cryoinjury were determined simultaneously on the mitochondrial function, motility, morphology and viability of ejaculated human sperm. METHOD: Rhodamine 123 (R123) uptake (% of sperm) and stain intensity were used to determine sperm mitochondrial activity before and after cryopreservation from the semen of 50 men attending for infertility investigation. Morphology was assessed using Tygerberg's strict criteria and viability was assessed by eosin Y. Sperm motility was measured using computer-assisted semen analysis (CASA). RESULTS: Freeze-thawing caused a 37% (P = 0.001) reduction in normal morphological forms of sperm. All CASA sperm motility parameters except amplitude of lateral head displacement were similarly reduced. R123 uptake and intensity within sperm mitochondria decreased by 36 and 47% respectively (both P = 0.001). In addition, there was a similar significant decrease (31%, P = 0.001) in the viability of the sperm. CONCLUSIONS: Sperm morphology, motility, mitochondrial activities and viability are equally susceptible to cryopreservation-induced damage. R123 intensity is a novel and robust indicator of mitochondrial function before and after such trauma. PMID- 11870123 TI - Chlamydia antibody testing in screening for tubal factor subfertility: the significance of IgG antibody decline over time. AB - BACKGROUND: Chlamydia (C.) trachomatis antibody testing in screening for tubal factor subfertility is limited by false negative results, i.e. negative Chlamydia antibody tests in patients with tubal pathology at laparoscopy. The present study was performed to determine whether decline in C. trachomatis IgG antibodies over time is responsible for those false negative results. METHODS: A total of 39 women with an initial titre of > or =64 were re-studied after 4-7 years. A new serum sample was collected from each patient. The initial and the second serum sample were tested for C. trachomatis IgG antibodies using a micro immunofluorescence assay (MIF). A species-specific enzyme-linked immunosorbent assay (ELISA) was used to validate the MIF test results. All patients filled out a questionnaire to determine risk factors for renewed C. trachomatis infection between the initial and second serum sample. RESULTS: Seven of the 39 patients (18.0%) showed a decline (>2 titre steps) in IgG antibodies by MIF over a period of 4-7 years, but IgG antibodies never became undetectable. In the 7/39 patients who showed a decline by MIF, signal/cut-off values by ELISA did not change. CONCLUSION: A decline in IgG antibody titre is not a significant cause of false negative Chlamydia antibody test results. PMID- 11870125 TI - The rate of aneuploidy is altered in spermatids from infertile mice. AB - BACKGROUND: It is now possible for infertile males to father their own genetic children through the technique of ICSI. This prospect has consequently prompted several investigations into the quality of sperm being retrieved from infertile males. One potential risk is the use of aneuploid sperm or spermatids, which might then be transferred to the fertilized oocyte. METHODS: In this investigation, aneuploidy of spermatids was assessed through immunocytochemistry using antibodies directed against chromosome centromeric regions and complexes. Three different types of infertile male mice with phenotypes closely resembling those described in human non-obstructive azoospermia [PP1cgamma-deficient mice, CREM-deficient mice and C57BL/6J.MAC-17(0--23) mice] were examined for chromosome numbers by counting the number of kinetochores in round spermatids using a CREST antiserum. RESULTS: PP1cgamma(-/-) and CREM(-/-) spermatids from infertile mice showed highly significant elevated levels in the rate of aneuploidy compared with wild-type animals (P < 0.0001). Thus infertile males with independent genetic mutations resulting in different histopathologies showed a high risk in the level of aneuploidy in their spermatids. CONCLUSIONS: These results suggest that impaired spermatogenesis may lead to production of aneuploid gametes. Analysis of aneuploidy in gametes from infertile men, coupled with appropriate genetic counselling, is recommended prior to ICSI. PMID- 11870126 TI - Polymorphisms of glutathione S-transferase M1 and male infertility in Taiwanese patients with varicocele. AB - BACKGROUND: To examine glutathione S-transferase M1 (GST M1) gene polymorphism and male infertility in Taiwanese patients with varicocele, 80 young male patients with varicocele (group 1), 62 young male patients with subclinical varicocele (group 2) and 60 normal young males (group 3) were recruited in this study. METHODS: GST M1 null homozygous genotype [GST M1-] and the occurrence of a 4977 bp deletion of sperm mitochondrial DNA (mtDNA) were determined by polymerase chain reaction. The 8-hydroxy-2'-deoxyguanosine (8-OHdG) content of sperm DNA was measured by high-performance liquid chromatography. RESULTS: The frequencies of GST M1- genotype were 43.8, 41.9 and 45% for patients in groups 1, 2 and 3 respectively. In group 1 patients with GST M1- genotype, the frequency of the presence of the 4977 bp deletion in sperm mtDNA (54.3%) was significantly higher than that of the patients without the 4977 bp deletion in sperm mtDNA (45.7%, OR: 2.63, P = 0.04). Patients of groups 1 and 2 with GST M1- genotype had significantly higher 8-OHdG content in sperm DNA and lower protein thiols and ascorbic acid in seminal plasma than those with GST M1+ genotype. CONCLUSION: GST M1- genotype predisposes to increased oxidative damage to sperm of patients with varicocele. PMID- 11870127 TI - Seminal antibodies to human 60kd heat shock protein (HSP 60) in male partners of subfertile couples. AB - BACKGROUND: Heat shock proteins (HSP) are essential mammalian and bacterial stress proteins. At the cellular level, they act as chaperones, have important regulatory functions, and are considered to be an essential factor for reproduction. Scarce information exists on the role of sensitization to HSP and the potential role in the aetiology of male infertility. METHODS: The potential association of immunoglobulin (Ig)A antibodies (Ab) to the human 60 kDa heat shock protein (HSP 60) with several parameters of subclinical male genital tract infection/inflammation and with semen quality and sperm fertilizing capacity was analysed in a prospective study. IgA Ab to human HSP 60 were determined in seminal plasma of 202 randomly chosen male partners of subfertile couples with a median duration of infertility of 4 years (range 1-15 years), who were asymptomatic for genital tract infection. After medical history and clinical examination, a comprehensive evaluation of semen quality, in aliquots of the same ejaculates used for HSP Ab determination, included: sperm analysis; local antisperm antibody (ASA) screening; standardized sperm-cervical mucus (CM) penetration testing; immunocytochemical round cell differentiation to determine seminal leukocyte counts; evaluation of complement fraction C(3) and of some pro inflammatory cytokines; and microbial screening. Subsequent fertility was recorded after 6 months. RESULTS: The presence of HSP 60 IgA Ab in seminal fluid (total positive 6.9%) was significantly associated with leukocytospermia, the presence of C(3), and also with high interleukin (IL) levels in seminal plasma. HSP 60 Ab were not related to the bacterial colonization of ejaculates. There was no association of seminal IgA Ab to human HSP 60 with semen quality, determined with microscopical semen analysis, nor with local IgG- or IgA-class ASA. There was no relationship with sperm intrinsic motility and duration of motility in the sperm CM-penetration test, nor with sperm fertilizing capacity. CONCLUSIONS: The combined presence of IgA Ab to human 60 kDa HSP, leukocytes and other established infection/inflammation markers in semen might suggest a potential role of the immune response to heat shock proteins (HSP) in cases of silent male genital tract infection, but the results do not indicate a marked relationship of HSP 60 Ab in seminal fluid with standard parameters of semen quality. PMID- 11870128 TI - Relationship between time period after vasectomy and the reproductive capacity of sperm obtained by epididymal aspiration. AB - BACKGROUND: It is not well defined whether the elapsed time after vasectomy has any influence on the outcome of IVF-ICSI using epididymal sperm. We analysed retrospectively the results of 151 ICSI cycles in which sperm of vasectomized men were used at different time periods after vasectomy. METHODS: Oocytes were obtained after a desensitizing ovarian stimulation protocol using GnRH agonist in association with recombinant FSH and HCG. Sperm were retrieved by percutaneous epididymal sperm aspiration. The cycles were split into three groups: < or =10 years after vasectomy (group 1, n = 47), 11-19 years after vasectomy (group 2, n = 79), and > or =20 years after vasectomy (group 3, n = 25). RESULTS: As might be expected, the mean age of men differed in the three groups (group 3 > group 2 > group 1), and the mean age of the women was also significantly higher in group 3 than in groups 1 and 2, although no differences were described between groups 2 and 3. All other laboratory and clinical parameters were similar in the three groups. Ongoing pregnancy and implantation rates (34, 25, 8% and 22, 15, 6% respectively) decreased significantly from group 1 to group 3. CONCLUSION: Pregnancy and implantation rates after ICSI with sperm from vasectomized men are negatively correlated with the time interval from vasectomy, which cannot be explained purely by male or female ageing. PMID- 11870129 TI - Transfer at the blastocyst stage of embryos derived from testicular round spermatid injection. AB - BACKGROUND: Intracytoplasmic injection of testicular round spermatids has been suggested as a salvage treatment in couples when testicular sperm extraction does not yield any mature sperm. However, the success of the procedure is debatable, and controversy surrounds issues such as the presence and (if present) identification of spermatids in testicular tissue. Progression rate to the blastocyst stage of spermatid-derived embryos appears to be low. METHODS: In this study, we investigated the feasibility and outcome of blastocyst stage embryo transfer after round spermatid injection (ROSI). ROSI was undertaken in 58 couples who did not yield mature or elongated sperm to testicular sperm extraction. RESULTS: The incidence of blastocyst formation from two pronuclear oocytes was 7.6%. A total of 16 blastocysts were transferred in 12 patients (20.7%). None of the patients conceived. CONCLUSIONS: The results of this study indicate that the blastocyst stage is reached by only very few ROSI-derived embryos and these embryos do not implant. PMID- 11870130 TI - Births after vitrification at morula and blastocyst stages: effect of artificial reduction of the blastocoelic cavity before vitrification. AB - BACKGROUND: In 1996, with the introduction of sequential media, we set up a programme of cryopreservation of supernumerary morulae (day 4) and blastocysts (day 5) using a vitrification procedure. Our results showed that the efficiency of the vitrification method was dependent on the stage of embryo development and was negatively correlated with the expansion of the blastocoele. We postulated that a large blastocoele might disturb cryopreservative potential due to ice crystal formation during the cooling step. We analysed therefore the effectiveness of reducing before vitrification the volume of the blastocoelic cavity. METHOD: Day 4 and day 5 embryos were vitrified in 40% ethylene glycol-18% Ficoll and 0.3 mol/l sucrose before plunging the straws directly into liquid nitrogen. Artificial shrinkage of the blastocyst was achieved after pushing a needle into the blastocoele cavity until it contracted. RESULTS: The survival rate post-thawing of day 4 and intact day 5 embryos was correlated with the volume of the blastocoele. In the control group only 20.3% blastocysts or expanded blastocysts survived as compared with 54.5 and 58.5% with morulae and early blastocyst respectively. After puncturing the blastocoelic cavity, an increase in the survival rate of up to 70.6% was noted. The pregnancy rates were improved after the artificial shrinkage procedure (20.5%) compared with the control intact blastocyst group (4.5%) (not significant). After reduction of the blastocoelic cavity, a significant increase in the implantation rate per vitrified blastocyst was observed (12.0 versus 1.4% P < 0.01). CONCLUSIONS: Our results showed that survival rates in cryopreserved expanded blastocysts could be improved by reducing the fluid content. This was presumably because mechanical damage caused by ice crystal formation was avoided. These observations should be considered when establishing a strategy and a protocol for cryopreservation of day 5 embryos. PMID- 11870131 TI - DNA fingerprinting of sister blastomeres from human IVF embryos. AB - BACKGROUND: Previously published single cell DNA fingerprinting systems have been plagued by high rates of allele drop-out (ADO) and preferential amplification (PA) preventing clinical application in preimplantation genetic diagnosis. METHODS: Tetranucleotide microsatellite markers with high heterozygosity, known allelic size ranges and minimal PCR stutter artefacts were selected for chromosomes X, 13, 18 and 21 and optimized in a multiplex fluorescent (FL)-PCR format. FL-PCR products were analysed using the ABI Prism 377 DNA sequenator and Genescan software. Validation of the DNA fingerprinting system was performed on single diploid (n = 50) and aneuploid (n = 25) buccal cells and embryonic blastomeres (n = 21). RESULTS: The optimized pentaplex PCR DNA fingerprinting system displayed a high proportion of successful amplifications (>91%) and low ADO and PA (<6%) when assessed on 50 human buccal cells. DNA fingerprints of single cells from a subject with Down's syndrome detected the expected tri allelic pattern for the chromosome 21 marker, confirming trisomy 21. In a blind study on 21 single blastomeres, all embryos were identifiable by their unique DNA fingerprints and shared parental alleles. CONCLUSIONS: A highly specific multiplex FL-PCR based on the amplification of five highly polymorphic microsatellite markers was developed for single cells. This finding paves the way for the development of a more complex PCR DNA fingerprinting system to assess aneuploidy and single gene mutations in IVF embryos from couples at genetic risk. PMID- 11870132 TI - Failure to infect embryos after virus injection in mouse zygotes. AB - BACKGROUND: The intracytoplasmic injection of sperm raises the problem that viral elements may be transported into the oocyte by the spermatozoon or the surrounding medium. It also raises questions about how the developing zygote will behave. METHODS: We used the murine model to microinject murine cytomegalovirus (MCMV) into the zygote ooplasm and followed the changes in these microinjected zygotes in vivo and in vitro over time. RESULTS: 80% of zygotes microinjected with viral suspension, and 80% injected with medium alone, survived. Although MCMV DNA was detected in 56% of injected embryos, up until the blastocyst stage, the mice born from these injected zygotes developed normally and did not contain MCMV DNA. When embryonic stem cells were co-incubated with MCMV and then transferred into healthy blastocysts, the offspring were normal and did not contain any MCMV DNA. CONCLUSIONS: Our observations suggest that even if MCMV DNA persists from the zygote to the blastocyst stage, its presence has no detrimental effect on pre-implantation or post-implantation development. PMID- 11870134 TI - Polycystic ovaries in childhood: a common finding in daughters of PCOS patients. A pilot study. AB - BACKGROUND: Polycystic ovarian syndrome (PCOS) is a controversial endocrine pathology and, recently, it has been suggested that the condition is hereditary. The aim of this study was to prospectively determine in daughters of patients with PCOS, by ultrasonographic (US) and colour Doppler analyses, the incidence of polycystic ovaries and search any correlation with specific hormonal parameters. METHODS: Fifteen prepubertal offspring (Group I) of patients with PCOS and 10 normal control prepubertal girls (Group II) were submitted to clinical, auxological, and basal hormonal assay. In addition all patients were submitted to US and colour Doppler ovarian and uterine evaluation. RESULTS: Among Group I girls the prevalence of polycystic ovaries was 93%, whereas no subjects among Group II had polycystic ovaries. The ovarian volume (2.76 +/- 1.21 ml versus 0.87 +/- 0.46 ml; P < 0.001) and the number of small sized follicles (5.36 +/- 2.2 versus 0.75 +/- 0.92; P < 0.001) were significantly higher in Group I than Group II patients. In addition, a normal stromal score and an absent stromal vascularization was observed in the control group. The hormone levels did not differ between the two groups. CONCLUSION: In conclusion we speculate that polycystic ovaries in childhood may be considered a sign of genetic predisposition to PCOS and that environmental factors may express the adult clinical and hormonal presentation of the syndrome. PMID- 11870133 TI - Doppler ultrasound examination of uterine arteries on the day of oocyte retrieval in patients with uterine fibroids undergoing IVF. AB - BACKGROUND: There is no consensus as to whether uterine fibroids have any adverse effects on the outcomes of assisted reproduction treatment. This prospective study compared implantation/pregnancy rates of women with and without fibroids undergoing IVF-embryo transfer and measured uterine blood flow indices of the fibroid group. METHODS: Patients who had fibroids that, during transvaginal scanning, were found to be not distorting the endometrial lining were placed in the fibroid group, while patients with normal uteri were controls. Those with previous myomectomy or pedunculated subserosal fibroids only were excluded. All received a standard ovarian stimulation regimen. Doppler ultrasound examinations of uterine arteries were carried out in the fibroid group prior to oocyte retrieval. RESULTS: Similar implantation/pregnancy rates, multiple pregnancy rates and pregnancy outcomes were noted in both groups. In the fibroid group, significantly lower pulsatility index (PI) and resistance index (RI) of the right uterine artery and the average of right and left uterine arteries were found in those failing to conceive than in those patients who subsequently conceived (P < 0.001). CONCLUSION: The presence of fibroids not distorting the endometrial lining does not adversely affect implantation and pregnancy rates during IVF embryo transfer. Significantly lower uterine artery PI and RI were found in non pregnant women with fibroids than in their pregnant counterparts. PMID- 11870135 TI - Embryotoxicity of peritoneal fluid in women with endometriosis. Its relation with cytokines and lymphocyte populations. AB - BACKGROUND: The goals of the present work were to study the embryotoxic effects of peritoneal fluid (PF) in women with or without endometriosis, and to relate any embryotoxicity to the severity of endometriosis, infertility or achievement of pregnancy, cytokine concentrations and lymphocyte populations. METHODS: Sixty six consecutive women of reproductive age, 54 with endometriosis (21 infertile) and 12 infertile without endometriosis, and another 12 fertile women as control group, were included in this study. They all underwent laparoscopy or laparotomy in the second half of the cycle, and PF was collected from the pouch of Douglas. The embryotoxicity of the PF was assessed by means of a mouse embryo assay, and expressed as the number of embryos that did not reach blastocyst stage. Cytokines and lymphocyte populations present in PF were also studied and correlated with embryotoxicity. RESULTS: PF embryotoxicity was increased in women with endometriosis, but there was little correlation with the severity of the disease. However, although a clear relationship to the presence of infertility was not found, embryotoxicity appeared to be lower in those infertile patients with endometriosis who later became pregnant. We found a significant increase in embryotoxicity in the presence of high cytokine concentrations, especially with interleukin-6, and less so with interleukin-8 (P < 0.05). No good correlation was observed with lymphocyte populations, but CD56 (NK) cells were significantly increased in the PF of women with endometriosis. In general, the correlations for embryotoxicity were better when PF was diluted at 20% (91.4 +/- 17 versus 68.1 +/ 31, P < 0.01). CONCLUSIONS: These results suggest that alteration in the production of cytokines in the PF, especially IL-6, besides contributing to the endometriosis and its evolution, probably increases embryotoxicity. However, no correlation was found between the latter and associated infertility. PMID- 11870136 TI - Pregnancy rates following ablative laparoscopic surgery for endometriomas. AB - BACKGROUND: In this prospective, cohort study we present the cumulative pregnancy rate following ablative laparoscopic surgery in patients with endometriomas. METHODS: The cyst was mobilized, fenestrated, and the capsule treated with the potassium-titanyl-phosphate (KTP) laser or bipolar diathermy. Pre- and post operative transvaginal ultrasound scans were performed, and a detailed fertility history recorded. RESULTS: There were 39 women (38 intention to treat as a single procedure) who had been trying to conceive for >12 months. The mean age of the patients was 33.8 years (range 20-43), and there were 42 cysts (three bilateral) with a mean diameter of 4.8 cm (range 2-25). The mean revised American Fertility Society score was 64.9 (range 22-124), and 29 (74.4%) patients had stage IV disease. Seven patients (18%) had previously had a live birth, and 17 (43.6%) had undergone assisted conception in the past. The cumulative pregnancy rate was 15/38 (39.5%). The pregnancy rate in patients with stage IV disease was 11/28 (39.3%). There were no major complications. CONCLUSIONS: Our results indicate that laparoscopic cyst fenestration and capsule ablation is a safe and effective treatment for improving fertility. PMID- 11870137 TI - In-vivo angiogenesis and progestogens. AB - BACKGROUND: Progestogens are used clinically for contraception, to control excessive menstrual bleeding, and to prevent estrogen-induced endometrial hyperplasia. A significant problem with progestogen-only methods of contraception is the induction of breakthrough bleeding. METHODS: The effects of different progestogens on angiogenesis were examined using two approaches. The mouse sponge angiogenesis assay employed direct delivery of the dose ranges achieved therapeutically. The angiogenic response to long-term intrauterine levonorgestrel exposure, compared with unexposed premenopausal endometrium, was also studied. RESULTS: In the mouse sponge assay, norethisterone and medroxyprogesterone acetate stimulated angiogenesis at all doses, but was dose-dependent for levonorgestrel and nomegestrol. Levonorgestrel stimulated angiogenesis in the dose range 100 pmol/l to 10 nmol/l, but not at higher doses. In contrast, nomegestrol acetate stimulated angiogenesis at high, but not low, doses. Expression of acidic and basic fibroblast growth factors, thymidine phosphorylase, vascular endothelial growth factor and adrenomedullin were unaltered in levonorgestrel-exposed endometrium compared with premenopausal controls. Vascular density was increased but endothelial proliferation reduced in levonorgestrel-exposed endometrium. CONCLUSIONS: This is the first report of the direct effects of a wide range of doses of different progestogens on angiogenesis; results suggest that vascular targeting may be an effective strategy to deal with progestogen-induced abnormal bleeding. PMID- 11870138 TI - Maternal serum beta-HCG and alpha-fetoprotein concentrations in singleton pregnancies following assisted reproduction. AB - BACKGROUND: The reason for the elevated levels of HCG in assisted reproduction pregnancies remains unknown. Our hypothesis was that this increase is caused by the ovarian superovulation therapy. METHODS: We compared the beta-HCG and alpha fetoprotein (AFP) multiples of the median (MoM) in singleton pregnancies after IVF or ICSI with those achieved by frozen embryo transfer (FET) in spontaneous cycles. RESULTS: The HCG and AFP MoMs (plus minus SEMs) of 59 FET pregnancies were compared with 144 IVF (including 48 ICSI) pregnancies. The maternal HCG of pregnancies following ovarian stimulation was 1.31 plus minus 0.08 MoM compared with 1.35 plus minus 0.12 MoM in the unstimulated ones. The values for AFP were 1.06 plus minus 0.05 versus 1.11 plus minus 0.05 respectively. No significant differences could be observed between pregnancies following stimulated IVF/ICSI and unstimulated FET cycles. CONCLUSIONS: Our results show that second trimester maternal serum HCG is also elevated in singleton pregnancies following spontaneous FET cycles. The increased maternal serum HCG in IVF pregnancies is thus not related to superovulation therapy. Because of the elevated maternal serum HCG levels, serum screening cannot be performed reliably in pregnancies following assisted reproduction technology. Ultrasonographic detection of the nuchal translucency is unaffected and should be used for this group of women undergoing assisted reproduction. PMID- 11870139 TI - The "vanishing embryo" phenomenon in an oocyte donation programme. AB - BACKGROUND: We studied the incidence of vanishing embryos (VE) in pregnancies achieved by oocyte donation and evaluated the obstetric and perinatal complications. METHOD: A retrospective study was carried out based on a chart review of 399 patients with multiple pregnancies from our oocyte donation programme. We defined vanishing phenomenon as the early resorption, in the first trimester, of one or more embryos in a multiple gestation, after confirming embryonic heart activity by transvaginal ultrasound. RESULTS: Vanishing embryo was observed in 75 patients (18.8%). In 60 patients (80%) this phenomenon occurred before the ninth gestational week. A higher incidence of VE was observed in patients who initially showed a higher number of gestational sacs (P < 0.03). Vaginal bleeding in the first trimester was significantly higher in patients with VE (P < 0.005). Miscarriage rate was similar in pregnancies with and without VE (P = NS). The incidence of pregnancy induced hypertension was decreased in the group with VE (P < 0.03). Preterm spontaneous rupture of membranes occurred more frequently in pregnancies with VE (P < 0.05). However, gestational age at delivery was similar in the group with VE and the controls. CONCLUSIONS: The high incidence of VE in pregnancies achieved by oocyte donation should be considered when counselling patients with high order multiple gestations. PMID- 11870140 TI - Cytokine imbalance in pregnancies with fetal chromosomal abnormalities. AB - BACKGROUND: The aim of the present study is to investigate the levels of some of the cytokines which may be involved in the mechanisms leading to the impairment of placental perfusion and to the onset of uterine contractions in pregnancies with fetal genetic abnormalities compared with controls. METHODS: The amniotic fluid and maternal plasma levels of interleukin-6, interleukin-8 and tumour necrosis factor-beta in patients with fetal chromosomal abnormalities were measured, as well as in euploid pregnancies in the seventh week of gestation. RESULTS: An increase of interleukin-6 (P = 0.034) and a decrease of interleukin-8 (P < or =0.0001) in amniotic fluid, and a decrease of interleukin-6 in the maternal plasma (P = 0.026) was shown in pregnancies with fetal chromosomal abnormalities. A positive correlation was observed between amniotic interleukin-8 and serum interleukin-6 in the presence of fetal aneuploidy (P < 0.006). CONCLUSION: Further investigations of cytokine imbalance in pregnancies with poor outcome as a consequence of genetic disorders rather than infection is warranted. PMID- 11870142 TI - Placenta percreta in week 10 of pregnancy with consecutive hysterectomy: case report. AB - Placenta percreta in early pregnancy is rare and has been documented in only a few cases. We report on a patient with abdominal pain in week 10 of pregnancy. Sonography revealed a defective embryonic development and the absence of a border line between trophoblast and myometrium, as well as invasive growth in the region of isthmocervical transition, so curettage was performed. Heavy bleeding at this stage made a hysterectomy necessary. Histological examination revealed a placenta percreta. Because of possible complications, the therapy of choice for a placenta percreta is a hysterectomy, as was performed in this case. PMID- 11870141 TI - A randomized, double-blind, placebo-controlled trial of intravenous immunoglobulin in the prevention of recurrent miscarriage: evidence for a therapeutic effect in women with secondary recurrent miscarriage. AB - BACKGROUND: Previous trials of intravenous immunoglobulin (IvIg) treatment of women with recurrent miscarriage (RM) have provided diverging results. This may be due to different inclusion criteria and suboptimal treatment protocols in some trials. METHODS: According to a computer-generated list, 58 women with at least four unexplained miscarriages were randomly assigned to receive infusions of high doses of IvIg or placebo starting as soon as the pregnancy test was positive. RESULTS: In the intention-to-treat analysis, a 45% live birth rate was found in both allocation groups. In patients with secondary RM, 50% in the treatment group and 23% in the placebo group had successful pregnancies (P = not significant). When data from the present and a previous placebo-controlled trial of the same treatment were combined, 15/26 (58%) of the patients with secondary RM in the treatment group versus 6/26 (24%) in the placebo group had successful outcomes (P < 0.02). Only 7% of the karyotyped abortuses were abnormal. CONCLUSIONS: IvIg may improve pregnancy outcome in patients with secondary RM. A new placebo-controlled trial focusing on this subgroup should be conducted to confirm the results. PMID- 11870143 TI - Prenatal diagnosis of female pseudohermaphroditism associated with bilateral luteoma of pregnancy: case report. AB - Female pseudohermaphroditism associated with luteoma of pregnancy (LP) is a rare condition characterized by varying degrees of masculinization of a female fetus. We describe a case, diagnosed at 13 weeks gestation. Transvaginal ultrasound at 5 weeks of gestation revealed a normal intrauterine gestational sac and an enlarged maternal right ovary. Re-examination at 13 weeks showed a fetus with male external genitalia. Cytogenetic investigation on amniotic fluid revealed a normal female karyotype 46,XX. Follow-up sonography confirmed the previous assignment of male external genitalia and a second amniocentesis was negative for the SRY gene. High levels of androgens were found in the maternal blood. A diagnosis of female pseudohermaphroditism associated with bilateral LP was made. A healthy girl was born by Caesarean section with complete masculinization of external genitalia (Prader V). Histology confirmed a bilateral LP. To the best of our knowledge this represents the first case of prenatal diagnosis of female pseudohermaphroditism associated with LP and demonstrates the feasibility of diagnosis by sonography from 13 weeks gestation. This is also the first case described of Prader V masculinization associated with LP. PMID- 11870144 TI - Supraphysiological concentrations of estradiol in menopausal women given repeated implant therapy do not adversely affect lipid profiles. AB - BACKGROUND: The effects of oral estrogen therapy (ERT) on lipids and metabolic parameters are well known, in contrast to the effects of subcutaneously administered estrogen, particularly high concentrations of estrogen. We examined metabolic parameters in cohorts of women with and without subcutaneous estrogen therapy with concomitant supra-normal concentrations of estradiol (SE). METHODS: Lipids and lipoprotein concentrations, low density lipid (LDL) subfractions, and activity of hepatic lipase (HL) were assessed in 30 menopausal women with SE and 19 control subjects not using ERT, matched for body mass index and age. RESULTS: Waist-hip ratio (WHR) and fasting insulin (FI) concentrations were lower in the SE group compared with the women not on ERT (P < 0.05). The concentrations of triglyceride and high density lipid (HDL) cholesterol were similar (P > 0.1), whereas total cholesterol (P < 0.05), LDL cholesterol (P < 0.05), and HL activity (P < 0.01) were lower in the SE group. Concentrations of the large, buoyant LDL I subfraction were significantly lower in the SE group (P < 0.05), but there was no difference in LDL III concentrations. CONCLUSIONS: Women with SE have similar triglyceride and HDL cholesterol levels but lower LDL cholesterol concentrations compared with post-menopausal women not taking ERT. The observations that the SE group showed reduced fasting insulin and WHR suggest that supra-normal circulating concentrations of estradiol, delivered subcutaneously, may beneficially influence insulin metabolism. PMID- 11870145 TI - The European study of assisted reproduction families: the transition to adolescence. AB - BACKGROUND: Findings are presented of the second phase of a European longitudinal study of families created by assisted reproduction. The present investigation reports on data obtained during the child's transition to adolescence. METHODS: A total of 102 IVF families, 94 donor insemination (DI) families, 102 adoptive families, and 102 families with a naturally conceived child were compared on standardized interview and questionnaire measures of parenting and children's psychological well-being. RESULTS: The assisted reproduction families were similar to the adoptive and natural conception families for many of the measures of the quality of parent-child relationships. To the extent that differences were found between the assisted reproduction families and the other family types, these reflected mainly more positive functioning among the assisted reproduction families, with the possible exception of the overinvolvement with their children of a small proportion of assisted reproduction mothers and fathers. The assisted reproduction children were functioning well and did not differ from the adoptive or naturally conceived children on any of the measures of psychological adjustment. However, only 8.6% had been told about their genetic origins. CONCLUSIONS: IVF and DI families with an early adolescent child appear to be functioning well. PMID- 11870146 TI - Ovarian artery resistance index as a marker of pituitary suppression. PMID- 11870147 TI - Uterine contractility decreases at the time of blastocyst transfer. PMID- 11870148 TI - Characterization of human sperm. PMID- 11870151 TI - Radiation dose escalation in combined-modality therapy for esophageal cancer. PMID- 11870152 TI - Substitute "prostate cancer" for "neuroblastoma"? PMID- 11870153 TI - Cardiac dysfunction in the trastuzumab clinical experience. PMID- 11870154 TI - Is there a "best" choice of second-line agent in the treatment of recurrent, potentially platinum-sensitive ovarian cancer? PMID- 11870155 TI - Recurrent ovarian cancer: evidence-based treatment. PMID- 11870156 TI - Risk management in BRCA1 and BRCA2 mutation carriers: lessons learned, challenges posed. PMID- 11870157 TI - INT 0123 (Radiation Therapy Oncology Group 94-05) phase III trial of combined modality therapy for esophageal cancer: high-dose versus standard-dose radiation therapy. AB - PURPOSE: To compare the local/regional control, survival, and toxicity of combined-modality therapy using high-dose (64.8 Gy) versus standard-dose (50.4 Gy) radiation therapy for the treatment of patients with esophageal cancer. PATIENTS AND METHODS: A total of 236 patients with clinical stage T1 to T4, N0/1, M0 squamous cell carcinoma or adenocarcinoma selected for a nonsurgical approach, after stratification by weight loss, primary tumor size, and histology, were randomized to receive combined-modality therapy consisting of four monthly cycles of fluorouracil (5-FU) (1,000 mg/m(2)/24 hours for 4 days) and cisplatin (75 mg/m(2) bolus day 1) with concurrent 64.8 Gy versus the same chemotherapy schedule but with concurrent 50.4 Gy. The trial was stopped after an interim analysis. The median follow-up was 16.4 months for all patients and 29.5 months for patients still alive. RESULTS: For the 218 eligible patients, there was no significant difference in median survival (13.0 v 18.1 months), 2-year survival (31% v 40%), or local/regional failure and local/regional persistence of disease (56% v 52%) between the high-dose and standard-dose arms. Although 11 treatment related deaths occurred in the high-dose arm compared with two in the standard dose arm, seven of the 11 deaths occurred in patients who had received 50.4 Gy or less. CONCLUSION: The higher radiation dose did not increase survival or local/regional control. Although there was a higher treatment-related mortality rate in the patients assigned to the high-dose radiation arm, it did not seem to be related to the higher radiation dose. The standard radiation dose for patients treated with concurrent 5-FU and cisplatin chemotherapy is 50.4 Gy. PMID- 11870158 TI - Phase II trial of chronomodulated infusion of high-dose fluorouracil and l folinic acid in previously untreated patients with metastatic colorectal cancer. AB - PURPOSE: To study tolerability and efficacy of an intensified chronomodulated schedule of fluorouracil (5-FU) and l-folinic acid (l-FA) as first-line treatment of metastatic colorectal cancer, 5-FU was given near individually determined dose limiting toxicity in a multicenter phase II trial. PATIENTS AND METHODS: One hundred patients (68 men and 32 women, median age 62 years, World Health Organization performance status less-than-or-equal 2) with previously untreated and inoperable metastases received chronomodulated daily infusion of 5-FU/l-FA (from 10:00 PM to 10:00 AM with peak at 4:00 AM). 5-FU dose was escalated from 900 to 1,100 mg/m(2)/d with fixed dose of l-FA at 150 mg/m(2)/d for 4 days every 14 days. RESULTS: 5-FU dose escalation was achieved in 66% of the patients. Grade 3 to 4 toxicities mainly consisted of nausea or vomiting (14% of patients and 1.5% of courses), hand-foot syndrome (38% of patients and 8% of courses), mucositis (26% of patients and 4% of courses), and diarrhea (21% of patients and 2.3% of courses). Objective response rate (ORR) was 41% (95% confidence interval, 31.5% to 50.5%). Twenty patients underwent metastases surgery; among these, 12 had a complete resection. Median progression-free survival was 7 months. Median survival was 17 months; 28% of the patients were alive at 2 years and 18.6% at 3 years. CONCLUSION: The ORR achieved with intensified chronomodulated delivery of 5-FU/l-FA was nearly twice as high as that earlier obtained by our cooperative group using less intensive 5-FU/FA chronotherapy. PMID- 11870159 TI - Irinotecan plus gemcitabine induces both radiographic and CA 19-9 tumor marker responses in patients with previously untreated advanced pancreatic cancer. AB - PURPOSE: This phase II, multicenter, open-label, single-arm study evaluated the efficacy and safety of irinotecan and gemcitabine as combination chemotherapy for previously untreated patients with unresectable or metastatic pancreatic cancer. PATIENTS AND METHODS: Patients received repeated 21-day cycles at starting doses of gemcitabine 1,000 mg/m(2) over 30 minutes followed immediately by irinotecan 100 mg/m(2) over 90 minutes, both given intravenously on days 1 and 8. Patients were evaluated for objective tumor response, changes in the serum tumor marker CA 19-9, time to tumor progression (TTP), survival, and safety. RESULTS: Forty-five patients were treated. Eleven patients (24%) had 50% or greater reductions in tumor area. These were confirmed one cycle later in nine patients (response rate, 20%; 95% confidence interval, 8% to 32%). Among 44 patients with baseline CA 19-9 determinations, CA 19-9 decreased during therapy in 22 patients (50%) and was reduced by 50% or more in 13 patients (30%). Median TTP was 2.8 months (range, 0.3 to 10.8 months). There were significant (P <.001) correlations between proportional changes in CA 19-9 and radiographic changes in tumor area with regard to extent of change (r =.67), timing of minimum on-study values (r =.85), and tumor progression (r =.89). Median survival was 5.7 months (range, 0.4 to 19.4+ months), and the 1-year survival rate was 27%. Severe toxicities were uncommon and primarily limited to grade 4 neutropenia (2%), grade 4 vomiting (2%), and grade 3 diarrhea (7%). CONCLUSION: Irinotecan/gemcitabine is a new combination that offers encouraging activity in terms of radiographic and CA 19-9 response and notable 1-year survival in pancreatic cancer. The regimen was well tolerated, with minimal grade 3 and 4 toxicities and excellent maintenance of planned dose-intensity. PMID- 11870160 TI - Age, sex, and racial differences in the use of standard adjuvant therapy for colorectal cancer. AB - PURPOSE: Dissemination of efficacious adjuvant therapies for resectable colorectal cancer has not been comprehensively described. Trends, patterns, and outcomes of adjuvant therapy for colorectal cancer, focusing on age, sex, and racial/ethnic differences, are reported. MATERIALS AND METHODS: Population-based random samples of patients diagnosed with colorectal cancer diagnosed in nine geographic areas were collected annually between 1987 and 1991 and in 1995 (n = 4,706). Data were obtained from medical record reviews. Multiple logistic regression was used to assess the use of standard adjuvant chemotherapy for colon and rectal cancers. The Cox proportional hazards model was used to assess 9-year mortality. RESULTS: From 1987 until 1995, the use of adjuvant therapy increased in all age groups. There was an increase starting in 1989 for colon and in 1988 for rectal cancer. Use of standard therapy was 78% for those younger than 55 years and 24% for those older than 80 years. White patients received standard therapy more frequently than African-Americans (odds ratio, 1.75; 95% confidence interval [CI], 1.09 to 2.83). All-cause and cancer-specific mortality exceeding 9 years were lower in those who received standard therapy (all-cause risk ratio [RR], 0.73; 95% CI, 0.61 to 0.88; cancer-specific RR, 0.87; 95% CI, 0.70 to 1.09). CONCLUSION: Standard adjuvant therapies for colorectal cancer disseminated into community practices during the 1990s. However, evidence exists of differential use of therapies by older patients and by African-Americans. The use of standard therapies in the general population is associated with lower mortality. Improved dissemination of standard adjuvant therapies to all segments of the population could help reduce mortality. PMID- 11870161 TI - Mutations of hMLH1 and hMSH2 in patients with suspected hereditary nonpolyposis colorectal cancer: correlation with microsatellite instability and abnormalities of mismatch repair protein expression. AB - PURPOSE: The relationship between germ-line mutations of hMSH2 and hMLH1, microsatellite instability (MSI), and loss of DNA mismatch repair (MMR) gene expression were studied to formulate an effective selection protocol for patients with suspected hereditary nonpolyposis colorectal cancer who should be offered genetic testing. PATIENTS AND METHODS: Patients eligible for germ-line analysis of hMLH1 and hMSH2 were selected. Tumor specimens were obtained to assess MSI and loss of MMR gene expression. RESULTS: Among 37 patients who participated in the study, two hMSH2 and two hMLH1 missense mutations (11%) were detected, none of which was found in a panel of 60 healthy volunteers. High MSI was found in five tumors (19%) and low MSI in 10 tumors (39%); 12 tumors (46%) were microsatellite stable. Four tumors demonstrated loss of hMLH1, and three tumors demonstrated loss of hMSH2 protein expression. CONCLUSION: No relationship was found between MMR gene mutations and MSI; low or no MSI was found in the four patients with germ-line mutations, and none of the five patients with high MSI demonstrated abnormalities of MMR genes. On the contrary, loss of hMLH1 or hMSH2 expression was found in the tumors from three of the four patients demonstrating germ-line mutations. These data suggest that germ-line mutations of the MMR gene can occur in people with MSI-negative tumors. Sensitive clinical criteria and the study of MMR gene expression may be useful to identify this subset of patients. PMID- 11870162 TI - Marginal decrease in mortality and marked increase in incidence as a result of neuroblastoma screening at 6 months of age: cohort study in seven prefectures in Japan. AB - PURPOSE: To determine the usefulness of 6-month screening for neuroblastoma. PATIENTS AND METHODS: The cumulative incidence rates (IRs) and cumulative mortality rates (MRs) of neuroblastoma in children younger than 60 months of age were analyzed for control (n = 713,025), qualitative screening (Qual Screen, n = 1,142,519), and quantitative screening (Quan Screen, n = 550,331) cohorts, and for Screened and Unscreened subgroups within screening cohorts. RESULTS: IRs (per 100,000) for infants aged 6 to 11 months were 1.12 in Control, 5.69 in Qual Screen (P <.0001), and 17.81 in Quan Screen (P <.0001); IRs for children aged 12 to 59 months were 7.29 in Control, 5.86 in Qual Screen (P =.28), and 6.36 in Quan Screen (P =.60). IRs for children aged 12 to 59 months in Unscreened or Screened subgroups remained at the same level. When patients diagnosed at younger than 6 months of age were excluded, the MR (per 100,000) under 60 months for Control was 4.21; those in Unscreened and Screened subgroups were 3.84 and 2.53 in Qual Screen (P =.30), and 3.20 and 1.97 in Quan Screen (P =.73), respectively; MRs between Control and Unscreened subgroups revealed no significant differences (P =.89 in Qual Screen, P =.85 in Quan Screen). CONCLUSION: Six-month screening resulted in a marked increase in incidence for infants with no significant decrease in incidence for children older than 1 year of age. A decrease in mortality was observed, but it was not significant. The usefulness of screening is questionable, because the decrease of mortality should be balanced against the adverse effect of overdiagnosis and the psychological burden on parents and children. PMID- 11870163 TI - Cardiac dysfunction in the trastuzumab clinical trials experience. AB - PURPOSE: This study sought to estimate cardiac dysfunction (CD) risk for patients receiving trastuzumab; to characterize observed CD by severity, treatment, and clinical outcome; to assess effects of baseline clinical risk factors on CD; and to assess effects of cumulative doses of anthracyclines and trastuzumab on CD. PATIENTS AND METHODS: A retrospective review of records for patients enrolled onto any of seven phase II and III trastuzumab clinical trials was performed. Predefined criteria were used for the diagnosis, and the New York Heart Association functional classification system was used to document CD severity. Product-limit estimates were used to summarize the cumulative anthracycline and trastuzumab doses at the time of CD onset. RESULTS: Patients treated with trastuzumab were found to be at an increased risk for CD. The incidence was greatest in patients receiving concomitant trastuzumab and anthracycline plus cyclophosphamide (27%). The risk was substantially lower in patients receiving paclitaxel and trastuzumab (13%) or trastuzumab alone (3% to 7%); however, most of these patients had received prior anthracycline therapy. CD was noted in 8% of patients receiving anthracycline plus cyclophosphamide and 1% receiving paclitaxel alone. Most trastuzumab-treated patients developing CD were symptomatic (75%), and most improved with standard treatment for congestive heart failure (79%). CONCLUSION: Trastuzumab is associated with an increased risk of CD, which is greatest in patients receiving concurrent anthracyclines. In most patients with metastatic breast cancer, the risk of CD can be justified given the improvement in overall survival previously reported with trastuzumab. PMID- 11870164 TI - Phase I and pharmacologic study of liposomal lurtotecan, NX 211: urinary excretion predicts hematologic toxicity. AB - PURPOSE: To determine the maximum-tolerated and recommended dose, toxicity profile, and pharmacokinetics of the liposomal topoisomerase I inhibitor lurtotecan (NX 211) administered as a 30-minute intravenous infusion once every 3 weeks in cancer patients. PATIENTS AND METHODS: NX 211 was administered by peripheral infusion. Dose escalation decisions were based on all toxicities during the first cycle as well as pharmacokinetic parameters. Serial plasma, whole blood, and urine samples were collected for up to 96 hours after the end of infusion, and drug levels were determined by high-performance liquid chromatography. RESULTS: Twenty-nine patients (16 women; median age, 56 years; range, 39 to 74 years) received 77 courses of NX 211 at dose levels of 0.4 (n = 3), 0.8 (n = 6), 1.6 (n = 3), 3.2 (n = 6), 3.8 (n = 6), and 4.3 mg/m(2) (n = 5). Neutropenia and thrombocytopenia were the dose-limiting toxicities and were not cumulative. Other toxicities were mild to moderate. Nine patients had stable disease while undergoing treatment. The systemic clearance of lurtotecan in plasma and whole blood was 0.82 +/- 0.78 L/h/m(2) and 1.15 +/- 0.96 L/h/m(2), respectively. Urinary recovery (Fu) of lurtotecan was 10.1% +/- 4.05% (range, 4.9% to 18.9%). In contrast to systemic exposure measures, the dose excreted in urine (ie, dose x Fu) was significantly related to the percent decrease in neutrophil and platelet counts at nadir (P <.00001). CONCLUSION: The dose limiting toxicities of NX 211 are neutropenia and thrombocytopenia. The recommended dose for phase II studies is 3.8 mg/m(2) once every 3 weeks. Pharmacologic data suggest a relationship between exposure to lurtotecan and NX 211-induced clinical effects. PMID- 11870165 TI - Randomized controlled trial of single-agent paclitaxel versus cyclophosphamide, doxorubicin, and cisplatin in patients with recurrent ovarian cancer who responded to first-line platinum-based regimens. AB - PURPOSE: To assess the activity, efficacy, and tolerability of single-agent paclitaxel and a platinum-containing regimen in previously treated patients with recurrent ovarian cancer. PATIENTS AND METHODS: Patients who achieved complete remission with platinum-based regimens and whose disease recurred after a progression-free interval of more than 12 months were included in the study. Every 21 days, patients received paclitaxel 175 mg/m(2) intravenously (IV) over 3 hours or cyclophosphamide 500 mg/m(2), doxorubicin 50 mg/m(2), and cisplatin 50 mg/m(2) (CAP) IV. RESULTS: Between June 1992 and May 1995, 97 consecutive patients with assessable or measurable disease were randomized to paclitaxel (n = 50) or CAP (n = 47). The median number of cycles on each arm was six. Toxicities included grade 3/4 leukopenia (4% for paclitaxel v 34% for CAP), grade 3/4 neutropenia (13% v 36%), grade 1/2 myalgia (19% v 4%), allergic reactions (15% v 2%), and grade 2/3 nausea and vomiting (17% v 51%). Complete responses were achieved in 17% and 30% of patients receiving paclitaxel and CAP, respectively, and partial responses were achieved in 28% and 25%, respectively (P =.062). At a median follow-up time of 49 months, median progression-free intervals were 9 months for paclitaxel and 15.7 months for CAP (Cox analysis: hazards ratio [HR], 0.60; 95% confidence interval [CI], 0.37 to 0.97; P =.038); median overall survival times were 25.8 months for paclitaxel and 34.7 months for CAP (Cox analysis: HR, 0.58; 95% CI, 0.34 to 0.98; P =.043). CONCLUSION: Rechallenge with either single-agent paclitaxel or platinum-based chemotherapy is effective in this patient population. Preliminary results suggest that single-agent paclitaxel may not be as active as platinum-based chemotherapy in recurrent ovarian cancer. Larger randomized trials are needed. PMID- 11870166 TI - Retrospective analysis of carboplatin and paclitaxel as initial second-line therapy for recurrent epithelial ovarian carcinoma: application toward a dynamic disease state model of ovarian cancer. AB - PURPOSE: The majority of patients with epithelial ovarian cancer (EOC) who achieve a complete remission with front-line chemotherapy develop recurrent disease. Carboplatin and paclitaxel are used for patients with platinum-sensitive recurrent disease, although there is little information regarding the response and survival in unselected patients treated with this strategy. We sought to determine the outcomes for patients with EOC treated with carboplatin and paclitaxel at the time of first recurrence. In addition, we sought to define a new paradigm for disease transition in patients with EOC. PATIENTS AND METHODS: Eighty-nine patients were identified who had a complete response to front-line platinum-based chemotherapy for EOC, relapsed 6 months after completion of front line chemotherapy, and were treated with carboplatin and paclitaxel as second line therapy. RESULTS: Eighty-four cases were available for analysis of survival end points, and 66 were assessable for response. The median follow-up was 27 months. The overall response rate was 70%. The median progression-free interval for the cohort was 13 months (95% confidence interval [CI], 10.7 to 13.8 months). The 3-year survival rate was 72% (95% CI, 59.4 to 86.1%). Toxicity was limited, and no deaths from treatment were observed. Using this data, it is possible to construct a disease states model of EOC, which provides risk estimates for transitions between clinically distinct categories. CONCLUSION: Re-treatment with carboplatin and paclitaxel is effective as initial therapy in recurrent EOC. This should form the basis of a randomized trial to determine the best agents for initial treatment of relapse from EOC in potentially platinum-sensitive patients. PMID- 11870167 TI - Survival effect of maximal cytoreductive surgery for advanced ovarian carcinoma during the platinum era: a meta-analysis. AB - PURPOSE: To evaluate the relative effect of percent maximal cytoreductive surgery and other prognostic variables on survival among cohorts of patients with advanced-stage ovarian carcinoma treated with platinum-based chemotherapy. MATERIALS AND METHODS: Eighty-one cohorts of patients with stage III or IV ovarian carcinoma (6,885 patients) were identified from articles in MEDLINE (1989 through 1998). Linear regression models, with weighted correlation calculations, were used to assess the effects on log median survival time of the proportion of each cohort undergoing maximal cytoreduction, dose-intensity of the platinum compound administered, proportion of patients with stage IV disease, median age, and year of publication. RESULTS: There was a statistically significant positive correlation between percent maximal cytoreduction and log median survival time, and this correlation remained significant after controlling for all other variables (P <.001). Each 10% increase in maximal cytoreduction was associated with a 5.5% increase in median survival time. When actuarial survival was estimated, cohorts with < or = 25% maximal cytoreduction had a mean weighted median survival time of 22.7 months, whereas cohorts with more than 75% maximal cytoreduction had a mean weighted median survival time of 33.9 months--an increase of 50%. The relationship between platinum dose-intensity and log median survival time was not statistically significant. CONCLUSION: During the platinum era, maximal cytoreduction was one of the most powerful determinants of cohort survival among patients with stage III or IV ovarian carcinoma. Consistent referral of patients with apparent advanced ovarian cancer to expert centers for primary surgery may be the best means currently available for improving overall survival. PMID- 11870168 TI - Outcome of preventive surgery and screening for breast and ovarian cancer in BRCA mutation carriers. AB - PURPOSE: To prospectively determine the impact of genetic counseling and testing on risk-reduction strategies and cancer incidence in a cohort of individuals at hereditary risk for breast and ovarian cancer. PATIENTS AND METHODS: Two hundred fifty-one individuals with BRCA mutations were identified at a single comprehensive cancer center from May 1, 1995, through October 31, 2000. Uniform recommendations regarding screening and preventive surgery were provided in the context of genetic counseling. Patients were followed for a mean of 24.8 months (range, 1.6 to 66.0 months) using standardized questionnaires, chart reviews, and contact with primary physicians. RESULTS: Frequency of cancer surveillance by physical examinations and imaging studies increased after genetic counseling and testing. Twenty-one breast, ovarian, primary peritoneal, or fallopian tube cancers were detected after receipt of genetic test results. Among 29 individuals choosing risk-reducing mastectomy after testing, two were found to have occult intraductal breast cancers. Among 90 individuals who underwent risk-reducing salpingo-oophorectomy, one early-stage ovarian neoplasm and one early-stage fallopian tube neoplasm were found. Radiographic or tumor marker-based screening detected six breast cancers, five of which were stage 0/I, one early-stage primary peritoneal cancer, and three stage I or II ovarian cancers. Six additional breast cancers were detected by physical examination between radiographic screening intervals; four of these six tumors were stage I. No stage III or stage IV malignancies were detected after genetic testing. CONCLUSION: This study provides prospective evidence that genetic counseling and testing increased surveillance and led to risk-reducing operations, which resulted in diagnosis of early-stage tumors in patients with BRCA1 and BRCA2 mutations. PMID- 11870169 TI - T-cell/histiocyte-rich large B-cell lymphoma: a distinct clinicopathologic entity. AB - PURPOSE: Although it has proven difficult to delineate diagnostically reproducible and clinically relevant subgroups, the heterogeneity of diffuse large B-cell lymphomas (DLBCL) is widely acknowledged. In 1992, we reported on six cases that suggested that large B-cell lymphoma rich in stromal histiocytes and T cells may be identified as a distinct clinicopathologic entity within DLBCL. PATIENTS AND METHODS: An integrated clinicopathologic study of 40 cases of this DLBCL subtype is presented. RESULTS: Distinguishing a DLBCL rich in histiocytes and reactive T cells, designated T-cell/histiocyte--rich large B-cell lymphoma (THR-BCL), may be justified from a clinical point of view. The disease typically affects middle-aged male patients who usually present with advanced stage disease that is not adequately managed with current therapeutic strategies. Whereas proliferation fraction and p53 overexpression, in addition to the clinical variables incorporated in the International Prognostic Index (IPI), significantly correlate with response to treatment and survival in a univariate analysis, only the IPI score identifies relevant prognostic THR-BCL subpopulations in a multivariate model. The morphologic and immunophenotypic profile of the neoplastic B cells in THR-BCL suggests that they may originate from a germinal center ancestor. CONCLUSION: THR-BCL constitutes a distinct clinicopathologic entity that is characterized by an aggressive behavior. Experimental therapeutic strategies may be indicated to obtain a more favorable response to treatment in this disease. PMID- 11870170 TI - CD20 expression in Hodgkin and Reed-Sternberg cells of classical Hodgkin's disease: associations with presenting features and clinical outcome. AB - PURPOSE: CD20 can be expressed in Hodgkin and Reed-Sternberg (HRS) cells of classical Hodgkin's disease (HD), but its clinical significance remains controversial. Therefore, we correlated CD20 expression with presenting features and clinical outcome of untreated patients with classical HD. PATIENTS AND METHODS: Patients were eligible if they were previously untreated and human immunodeficiency virus-1 negative, had biopsy-proven classical HD, and if pretreatment paraffin-embedded tumor tissue was available. CD20 expression was determined by immunohistochemistry without knowledge of clinical outcome. A tumor was considered positive if any HRS cells expressed CD20, but other cutoffs for number of CD20-positive HRS were also investigated. RESULTS: We identified 598 patients whose median age was 30 years and of whom 55% were male. HRS cells expressed CD20 in 132 (22%) of 598 patients with classical HD. When any percentage of CD20 expression in HRS cells was used as a cutoff, the 5-year failure-free survival (FFS) for positive versus negative tumors was 86% versus 84%, respectively, for 302 patients treated with doxorubicin, bleomycin, vinblastine, and dacarbazine or equivalent regimens (P =.7 by log-rank test), 74% versus 77%, respectively, for 181 patients treated with mitoxantrone, vincristine, vinblastine, and prednisone and radiotherapy (P =.7 by log-rank test), 74% versus 84%, respectively, for 54 patients treated with MOPP (P =.4 by log-rank test), and 77% versus 88% for 53 patients treated only with radiotherapy (P =.5 by log-rank test). The 5-year FFS was not statistically different when cutoffs of 5% up to 50% for CD20-positive HRS cells were used. CONCLUSION: CD20 is expressed by HRS cells in 22% of patients with classical HD but is not associated with different FFS after treatment with equivalent regimens. PMID- 11870171 TI - Rituximab and CHOP induction therapy for newly diagnosed mantle-cell lymphoma: molecular complete responses are not predictive of progression-free survival. AB - PURPOSE: To evaluate the efficacy of rituximab and cyclophosphamide, doxorubicin, vincristine, and prednisone (CHOP) induction therapy in patients with newly diagnosed mantle-cell lymphoma (MCL). PATIENTS AND METHODS: From March 1997 to May 1999, 40 previously untreated patients with stage II through IV MCL were treated with six cycles of rituximab and CHOP chemotherapy in a phase II trial. Pretreatment and interval peripheral-blood (PB) and bone marrow (BM) specimens were also analyzed by polymerase chain reaction (PCR) for tumor-specific BCL 1/immunoglobulin H (IgH) translocations and clonal IgH rearrangements. Study end points included clinical and molecular response rates and long-term progression free survival (PFS). RESULTS: Forty-eight percent of patients achieved a complete response (CR)/CR unconfirmed (CRu), and 48% of patients obtained a partial response (PR). However, 28 of the 40 patients have already relapsed or developed progressive disease with a median PFS of 16.6 months. Twenty-five patients had PCR-detectable BCL-1/IgH or clonal IgH products in PB or BM at diagnosis. Nine of the 25 informative patients had no evidence of PCR-detectable disease in PB or BM after rituximab and CHOP therapy. However, patients who achieved molecular remissions in PB or BM had PFS similar to patients without molecular remissions (16.5 v 18.8 months, P =.51). CONCLUSION: Favorable clinical and molecular response rates associated with rituximab and CHOP chemotherapy do not translate into prolonged PFS in MCL. Nevertheless, rituximab and combination chemotherapy may transiently clear PB or BM of detectable tumor cells, prompting additional consideration of antibody-based in vivo purging in subsequent clinical trials. PMID- 11870172 TI - Improved outcome of allogeneic transplantation in high-risk multiple myeloma patients after nonmyeloablative conditioning. AB - PURPOSE: We present our experience with relapsed and recently diagnosed patients with high-risk multiple myeloma (MM) receiving immunosuppressive, nonmyeloablative melphalan (MEL)-based conditioning regimens (mini-allograft). PATIENTS AND METHODS: Thirty-one MM patients received allografts from HLA-matched siblings (n = 25) or unrelated donors (n = 6) using a mini-allograft. Seventeen had progressive disease (PD) and 14 had responsive disease (RD) (six with primary RD and eight with responsive relapse). Thirty patients had received one (n = 13) or two or more (n = 17) prior autologous transplantations (ATs). Median age was 56 years (range, 38 to 69 years). Twenty-one patients had chromosome 13 abnormality. Two patients were hemodialysis dependent. Blood and bone marrow grafts were administered to 28 and three patients, respectively. Donor lymphocyte infusions were given to 18 patients either to attain full donor chimerism (n = 6) or to eradicate residual disease (n = 12). RESULTS: By day 100, 25 (89%) of 28 patients were full donor chimeras, one was a mixed chimera, and two had autologous reconstitution. Acute graft-versus-host disease (GVHD) developed in 18 patients (58%), and 10 progressed to chronic GVHD (limited in six and extensive in four). At a median follow-up of 6 months, 19 (61%) of 31 patients achieved complete/near complete remission. Twelve patients (39%) have died: three of PD, three of early treatment-related mortality (TRM) (before day 100), and six of late TRM. Median overall survival (OS) was 15 months. At 1 year, there was a significantly longer event-free survival (86% v 31%, P =.01) and OS (86% v 48%, P =.04) when a mini-allograft was performed after one versus two or more prior ATs, respectively. When compared with historical MM controls (n = 93) receiving conventional allografts, early TRM was significantly lower (10% v 29%, P =.03), and OS at 1 year was better (71% v 45%; P =.08) in the mini-allograft MM patients. CONCLUSION: Mini-allograft induced excellent disease control in MM patients with high-risk disease, but is still associated with a significant GVHD. PMID- 11870174 TI - Quality-of-life-adjusted survival analysis of high-dose adjuvant interferon alpha 2b for high-risk melanoma patients using intergroup clinical trial data. AB - PURPOSE: High-dose adjuvant interferon alpha-2b (IFN alpha 2b) for high-risk melanoma is a 1-year regimen that improves relapse-free and overall survival but has significant toxicity. A quality-of-life--adjusted survival (QAS) analysis analysis of two cooperative group phase III trials, E1684 and E1690/S9111/C9190, was performed, incorporating patient values (utilities) for the toxicity of IFN alpha 2b treatment and melanoma recurrence. PATIENTS AND METHODS: Quality Adjusted Time Without Symptoms or Toxicity methodology was used with melanoma patient utilities and trial data to estimate the effect of IFN alpha 2b on QAS. The increase or decrease in QAS that patients could expect from treatment was estimated based on their utilities. Eleven utility predictor questions were tested to identify patients with utilities that result in decreased QAS. RESULTS: Using E1684 data, IFN alpha 2b would result in an increase in QAS for all sets of patient utilities. This benefit was significant (P <.05) for 16% of patients. Using E1690/S9111/C9190 data, 77% of patients would experience a benefit in QAS from IFN alpha 2b and 23% would experience a decrease in QAS; neither of these effects was statistically significant. Using utility predictors and the E1690/S9111/C9190 analysis, a decision rule was formulated that helps identify patients in whom IFN alpha 2b may detract from QAS. CONCLUSION: Most patients experienced improvement in QAS in both trials, but this benefit was statistically significant in only 16% of patients in E1684. Change in QAS depends more on the utility for IFN alpha 2b toxicity than on the utility for melanoma recurrence. Cancer patients probably have higher utilities for IFN alpha 2b toxicity than members of the general population and will tend to favor IFN alpha 2b treatment as a result. PMID- 11870173 TI - Incidence and prognostic significance of complete axillary downstaging after primary chemotherapy in breast cancer patients with T1 to T3 tumors and cytologically proven axillary metastatic lymph nodes. AB - PURPOSE: To determine the incidence and prognostic significance of eradication of cytologically proven axillary lymph node metastases in breast cancer patients treated with primary chemotherapy. PATIENTS AND METHODS: Between January 1985 and December 1994, 152 breast cancer patients with invasive T1 to T3 tumors and axillary metastases cytologically proven by fine-needle sampling underwent primary chemotherapy followed by lumpectomy or mastectomy, level I and II axillary lymph node dissection, and irradiation. We studied pathologic complete responses (pCRs) of axillary nodes and breast tumors, as well as predictors of distant metastases. RESULTS: Thirty-five patients (23%) had axillary pCRs, and 20 patients (13.2%) had pCRs of primary breast tumors. Scarff-Bloom-Richardson grade 3 tumors (P =.04) and a clinical response to chemotherapy > or = 50% (P =.003) were associated with negative axillary status at dissection. An initial tumor size < or = 3 cm (63 patients) was associated with pCR of the primary tumor (P =.02) but not with complete histologic clearance of axillary lymph nodes. The median length of follow-up was 75 months. In the univariate analysis, age greater than 40 years (P =.003), absence of residual nodal disease (P =.01), and pCR of the tumor (P =.05) were associated with better distant disease-free survival. Five-year distant disease-free survival rates were 73.5% +/- 14.9% among patients with no involved nodes at the time of surgery and 48.7% +/- 9.2% among patients with residual nodal disease. In the multivariate Cox regression analysis, parameters associated with poor distant disease-free survival were age < or = 40 years (P =.002), persistence of nodal involvement (P =.03), and S-phase fraction greater than 4% (P =.02). CONCLUSION: Our results suggest that axillary status is a better prognostic factor than response of the primary tumor to primary chemotherapy. PMID- 11870175 TI - Clinical factors associated with cancer-related fatigue in patients being treated for leukemia and non-Hodgkin's lymphoma. AB - PURPOSE: To describe fatigue severity, fatigue interference, and associated factors in hematologic malignancies. PATIENTS AND METHODS: Patients being treated for leukemia and non-Hodgkin's lymphoma (n = 228) completed the Brief Fatigue Inventory to rate fatigue severity and functional interference caused by fatigue. Data on patient demographics, Eastern Cooperative Oncology Group performance status, other physical symptoms, current treatments, and laboratory values were also collected. Descriptive statistics, bivariate correlation, and logistic regression were used for data analysis. RESULTS: Fifty percent of the sample reported severe fatigue, which was defined as a "fatigue worst" rating of 7 or greater. More patients with acute leukemia (61%) reported severe fatigue compared with those with chronic leukemia (47%) and non-Hodgkin's lymphoma (46%). Increased fatigue severity significantly compromised patients' general activity, work, enjoyment of life, mood, walking, and relationships with others. Fatigue severity was strongly associated with performance status, use of opioids, blood transfusions, gastrointestinal symptoms, and sleep disturbance items, as well as with low serum hemoglobin and albumin levels. Regression analysis indicated that nausea was the significant clinical predictor of severe fatigue (odds ratio, 13), and low serum albumin was the significant laboratory value predictor (odds ratio, 3.8). CONCLUSION: Disabling fatigue occurs with high frequency in hematologic malignancy, supporting a need to develop better methods of fatigue management. Better control of gastrointestinal and other symptoms may be of benefit. The mechanism and relationship between low albumin and severe fatigue needs to be investigated further, and longitudinal studies of the effects of treatment, host factors, and other symptoms are needed. PMID- 11870176 TI - Increasing single epirubicin doses in advanced soft tissue sarcomas. AB - PURPOSE: To evaluate the maximum-tolerated dose and the clinical efficacy of epirubicin in patients with advanced soft tissue sarcoma. PATIENTS AND METHODS: Sixty-one patients were treated at three different epirubicin dose levels: 140 mg/m(2) (six patients), 160 mg/m(2) (52 patients), and 180 mg/m(2) (three patients). Cycles were repeated every 3 weeks for a maximum of eight cycles. The first two dose levels proved to be feasible and safe without dose-limiting toxicity (DLT). Because the first three patients entering the third dose level experienced DLT, subsequent patients received the next lower dose level. RESULTS: The overall response rate was 44% (95% confidence interval, +/- 12%), with six complete (10%) and 21 partial (34%) responses. Responses seemed related to epirubicin dose level, because the response rate was 17%, 44%, and 100% for the three dose levels (chi(2) test for trend, P =.02). Median response duration, median time to progression, and median overall survival were 10, 8, and 15 months, respectively. Myelosuppression was the most frequent side effect, with grade 3 or 4 neutropenia occurring in 79% of the patients; 31% of patients were febrile. Nonhematologic toxicity was mainly grades 1 and 2. The mean epirubicin dose-intensity was 49 mg/m(2) per week. CONCLUSION: The third epirubicin dose level (180 mg/m(2)) was the maximum-tolerated dose. The recommended drug dose for clinical use is 160 mg/m(2) every 3 weeks with hematopoietic support. Single high dose epirubicin is effective as first-line treatment and should be preferentially used whenever a high response rate is important to allow the resection of an otherwise unresectable disease or whenever it might result in a significant symptomatic benefit. PMID- 11870177 TI - Phase III trial comparing a defined duration of therapy versus continuous therapy followed by second-line therapy in advanced-stage IIIB/IV non-small-cell lung cancer. AB - PURPOSE: To compare four cycles of therapy versus continuous therapy to determine the optimal duration of chemotherapy in advanced non-small-cell lung cancer (NSCLC). PATIENTS AND METHODS: Stage IIIB/IV NSCLC patients were randomized to arm A (four cycles of carboplatin at an area under the curve of 6 and paclitaxel 200 mg/m(2) every 21 days) or arm B (continuous treatment with carboplatin/paclitaxel until progression). At progression, all patients on both arms were to receive second-line weekly paclitaxel at 80 mg/m(2)/wk. The primary end points were survival and quality of life (QOL). RESULTS: Two hundred thirty patients were randomized. Fifty-seven percent of arm A patients completed four courses of therapy. In the 116 arm B patients, the median number of cycles delivered was four (range, zero to 19 cycles). Forty-two percent received five or more cycles; 18% received eight or more cycles. Overall response rates were 22% and 24% for arms A and B, respectively (P =.80). Median survival time and 1-year survival rates were 6.6 months and 28% for arm A and 8.5 months and 34% for arm B, respectively (log-rank P =.63). Rates of hematologic and nonhematologic toxicity were similar between the two arms, except for neuropathy. The rate of grade 2 to 4 neuropathy increased from 19.9% (95% confidence interval [CI], 13.6% to 26.2%) at cycle 4 to 43% (95% CI, 28.6% to 57.4%) at cycle 8. There were no differences in QOL. Only 45% of patients received second-line therapy (42% in arm A v 47% in arm B, P =.42). CONCLUSION: This study shows no overall benefit in survival, response rates, or QOL to continuing treatment with carboplatin/paclitaxel beyond four cycles in advanced NSCLC. PMID- 11870178 TI - Economic analysis of the TAX 317 trial: docetaxel versus best supportive care as second-line therapy of advanced non-small-cell lung cancer. AB - PURPOSE: To determine the cost-effectiveness (CE) of second-line docetaxel compared with best supportive care (BSC) in the TAX 317 trial, a randomized clinical trial of second-line chemotherapy in non-small-cell lung cancer. METHODS: A retrospective CE analysis of the TAX 317 trial was undertaken, evaluating direct medical costs of therapy from the viewpoint of Canada's public health care system. Costs were derived in 1999 Canadian dollars, and resource use was determined through prospective trial data. RESULTS: The incremental survival benefit in the docetaxel arm over BSC was 2 months (P =.047). The CE of docetaxel was $57,749 per year of life gained. For patients treated with docetaxel 75 mg/m(2), the CE was $31,776 per year of life gained. In univariate sensitivity analyses, CE estimates were most sensitive to changes in survival, ranging from $18,374 to $117,434 with 20% variation in survival at the recommended dose. The largest cost center in both arms was hospitalization, followed by the cost of drugs, investigations, radiotherapy, and community care. BSC patients had fewer hospitalizations than patients in the chemotherapy arm and were more often palliated at home. CONCLUSION: Although the decision to treat should not be based on economic considerations alone, our CE estimate of $31,776 per year of life gained (at the currently recommended dose of docetaxel) is within an acceptable range of health care expenditures, and the total costs of therapy are similar to those of second-line palliative chemotherapy for other solid tumors. PMID- 11870179 TI - Prognostic significance of molecular genetic aberrations on chromosome segment 11p15.5 in non-small-cell lung cancer. AB - PURPOSE: The assessment of prognosis and decisions on treatment for patients with non-small-cell lung cancer (NSCLC) are determined on the basis of disease stage and performance status. NSCLC frequently manifests loss of heterozygosity (LOH) at chromosome segment 11p15.5. Whether LOH at 11p15.5 is an independent prognostic variable has yet to be determined. PATIENTS AND METHODS: We developed five novel markers, which can be assessed by polymerase chain reaction and restriction enzyme digestion. LOH at 11p15.5 was assessed in 193 patients who underwent surgical resection for pathologic stage I and II of the disease. RESULTS: LOH at 11p15.5 was associated with poor survival (P =.021). Multivariate logistic regression analysis showed that after disease stage, performance status, weight loss, sex, age at diagnosis, and smoking history were controlled for, patients with LOH were two times more likely to die than those without LOH (relative risk [RR] = 2.01, P =.021). Cox regression analysis with disease stage and LOH revealed that the survival of patients with stage I disease and LOH was similar to the survival of patients with stage II disease, and it was significantly worse than the survival of stage I patients without LOH (RR = 2.38, P =.038). CONCLUSION: LOH in a 310-kb region on chromosome segment 11p15.5 that includes the gene for the regulatory subunit of the enzyme ribonucleotide reductase is highly predictive of poor survival from NSCLC. The future utility of analysis of the allelic status of this region may involve treatment decisions, such as the use of neoadjuvant and adjuvant chemotherapy for patients with stage I disease. PMID- 11870181 TI - Mathematical model to predict individual survival for patients with renal cell carcinoma. AB - PURPOSE: To develop a multivariate model and mathematical formula capable of calculating personalized survival for renal cell carcinoma (RCC) patients with clinically available variables. PATIENTS AND METHODS: A total of 477 patients out of 661 undergoing nephrectomy at the University of California Los Angeles between 1989 and 1999 were eligible for evaluation and formed the analyzed cohort for this retrospective study. Time to death was the primary end point assessed. Univariate analysis for 14 to 20 variables was conducted, followed by a multivariate Cox analysis. The variables that provided independent information as to the time of death for metastatic and nonmetastatic patients were coded and incorporated into a function based on the Nadas equation principle. RESULTS: For nonmetastatic patients, the significant variables in the multivariate analysis were Fuhrman's grade and Eastern Cooperative Oncology Group performance status. For the metastatic patients, Fuhrman's grade, 1997 classification T stage, number of symptoms, nodal involvement, and immunotherapy were independent predictors for survival. These variables, based on the Cox multivariate regression model, were implanted into an exponential Nadas equation. The expected survival predicted by use of the Nadas equations faithfully describes the actual survival based on Kaplan-Meier curves. CONCLUSION: We have developed mathematical equations for estimating survival after radical nephrectomy for RCC. The resulting formulas are capable of better tailoring survival estimates for a specific patient and are based on widely accepted clinical prognostic variables. On validation with external data, this type of representation can be used as a tool for the determination of personalized prognosis and may be useful for patient education and counseling. PMID- 11870180 TI - Phase III trial of fluorouracil, interferon alpha-2b, and cisplatin versus methotrexate, vinblastine, doxorubicin, and cisplatin in metastatic or unresectable urothelial cancer. AB - PURPOSE: Previously, we developed a novel biochemotherapy regimen combining interferon alpha-2b with fluorouracil and cisplatin (FAP). We now report the results of a prospective randomized trial comparing FAP with methotrexate, vinblastine, doxorubicin, and cisplatin (M-VAC), the standard chemotherapy regimen for locally advanced and metastatic urothelial cancer. The purpose of this study was to compare the response rates and overall survival of patients with metastatic or unresectable urothelial cancer treated with these two chemotherapy regimens. PATIENTS AND METHODS: Between October 1992 and September 1999, 172 previously untreated patients were registered and randomly assigned to treatment with either FAP or M-VAC. Patients were followed until their death. RESULTS: The pretreatment clinical characteristics of the groups were similar except for sex (P <.01). Sex did not affect prognosis or survival. The objective response rate for patients assigned to FAP was 42% (35 of 83 patients), with complete response observed in eight (10%) of 83 patients. Among the patients assigned to M-VAC, 51 (59%) of 86 had an objective response, with complete response observed in 21 (24%) of 86. The Kaplan-Meier estimate of median survival was 12.5 months for both groups. Both regimens were quite toxic, with more mucocutaneous toxicity in the FAP arm and more myelosuppression in the M-VAC arm. CONCLUSION: Although overall survival was not significantly different, patients assigned to M-VAC had a much better chance of responding to front-line therapy. Thus, FAP is very likely to be inferior to M-VAC and is certainly no less toxic. FAP cannot be recommended as part of the standard armamentarium for urothelial cancer. PMID- 11870182 TI - Promising survival for patients with newly diagnosed glioblastoma multiforme treated with concomitant radiation plus temozolomide followed by adjuvant temozolomide. AB - PURPOSE: Temozolomide is a novel oral alkylating agent with demonstrated efficacy as second-line therapy for patients with recurrent anaplastic astrocytoma and glioblastoma multiforme (GBM). This phase II study was performed to determine the safety, tolerability, and efficacy of concomitant radiation plus temozolomide therapy followed by adjuvant temozolomide therapy in patients with newly diagnosed GBM. PATIENTS AND METHODS: Sixty-four patients were enrolled onto this open-label, phase II trial. Temozolomide (75 mg/m(2)/d x 7 d/wk for 6 weeks) was administered orally concomitant with fractionated radiotherapy (60 Gy total dose: 2 Gy x 5 d/wk for 6 weeks) followed by temozolomide monotherapy (200 mg/m(2)/d x 5 days, every 28 days for six cycles). The primary end points were safety and tolerability, and the secondary end point was overall survival. RESULTS: Concomitant radiation plus temozolomide therapy was safe and well tolerated. Nonhematologic toxicities were rare and mild to moderate in severity. During the concomitant treatment phase, grade 3 or 4 neutropenia, thrombocytopenia, or both were observed in 6% of patients, including two severe infections with Pneumocystis carinii. During adjuvant temozolomide, 2% and 6% of cycles were associated with grade 3 and 4 neutropenia or thrombocytopenia, respectively. Median survival was 16 months, and the 1- and 2-year survival rates were 58% and 31%, respectively. Patients younger than 50 years old and patients who underwent debulking surgery had the best survival outcome. CONCLUSION: Continuous daily temozolomide and concomitant radiation is safe. This regimen of concomitant chemoradiotherapy followed by adjuvant chemotherapy may prolong the survival of patients with glioblastoma. Further investigation is warranted, and a randomized trial is ongoing. PMID- 11870183 TI - Phase II trial of temozolomide plus the matrix metalloproteinase inhibitor, marimastat, in recurrent and progressive glioblastoma multiforme. AB - PURPOSE: Novel therapies are needed for patients with recurrent glioblastoma multiforme (GBM). Because there is evidence that temozolomide (TMZ) has some activity in GBM and is well tolerated, and because of laboratory evidence that metalloproteinases are important in glioma cell invasion, the combination of TMZ and the matrix metalloproteinase inhibitor marimastat (MRM) in patients with recurrent GBM was studied. PATIENTS AND METHODS: Forty-four patients with recurrent GBM after standard radiotherapy were enrolled. For 19 patients, this therapy was their first chemotherapy after tumor progression after irradiation; 25 others had received chemotherapy previously. TMZ 150 to 200 mg/m(2) days 1 to 5 and MRM 50 mg days 8 to 28 was administered at 28-day intervals for two cycles; then patients were reevaluated. Treatment continued until progression of tumor or toxicity developed. RESULTS: Joint and tendon pain was the major therapy-related toxicity and was reported in 47% of patients. Five patients (11%) were removed from the study because of intolerable joint pain. For all patients, the progression-free survival (PFS) at 6 months was 39%. Median PFS was 17 weeks, median overall survival was 45 weeks, and 12-month PFS was 16%. CONCLUSION: The combination of TMZ and MRM resulted in a PFS at 6 months that exceeded the literature target by 29%. This drug combination met phase II study criteria; further study in recurrent patients with GBM might be warranted. Further study of therapy-induced joint pain is necessary. PMID- 11870184 TI - Phase II trial of murine (131)I-labeled antitenascin monoclonal antibody 81C6 administered into surgically created resection cavities of patients with newly diagnosed malignant gliomas. AB - PURPOSE: To assess the efficacy and toxicity of intraresection cavity (131)I labeled murine antitenascin monoclonal antibody 81C6 and determine its true response rate among patients with newly diagnosed malignant glioma. PATIENTS AND METHODS: In this phase II trial, 120 mCi of (131)I-labeled murine 81C6 was injected directly into the surgically created resection cavity of 33 patients with previously untreated malignant glioma (glioblastoma multiforme [GBM], n = 27; anaplastic astrocytoma, n = 4; anaplastic oligodendroglioma, n = 2). Patients then received conventional external-beam radiotherapy followed by a year of alkylator-based chemotherapy. RESULTS: Median survival for all patients and those with GBM was 86.7 and 79.4 weeks, respectively. Eleven patients remain alive at a median follow-up of 93 weeks (range, 49 to 220 weeks). Nine patients (27%) developed reversible hematologic toxicity, and histologically confirmed, treatment-related neurologic toxicity occurred in five patients (15%). One patient (3%) required reoperation for radionecrosis. CONCLUSION: Median survival achieved with (131)I-labeled 81C6 exceeds that of historical controls treated with conventional radiotherapy and chemotherapy, even after accounting for established prognostic factors including age and Karnofsky performance status. The median survival achieved with (131)I-labeled 81C6 compares favorably with either (125)I interstitial brachy-therapy or stereotactic radiosurgery and is associated with a significantly lower rate of reoperation for radionecrosis. Our results confirm the efficacy of (131)I-labeled 81C6 for patients with newly diagnosed malignant glioma and suggest that a randomized phase III study is indicated. PMID- 11870185 TI - Standardized uptake value of 2-[(18)F] fluoro-2-deoxy-D-glucose in predicting outcome in head and neck carcinomas treated by radiotherapy with or without chemotherapy. AB - PURPOSE: In patients with head and neck cancer enrolled onto a prospective study of positron emission tomography (PET), pretreatment 2-[(18)F] fluoro-2- deoxy-D glucose (FDG) uptake was evaluated as a predictor of local control and disease free survival (DFS) after treatment by radiotherapy (RT) with or without chemotherapy. PATIENTS AND METHODS: We studied 63 patients with carcinomas of the head and neck who had an FDG-PET scan before radical RT. Tumor FDG uptake was measured with the semiquantitative standardized uptake value (SUV). All patients but one were treated with accelerated or hyperfractionated RT schedules. Thirteen patients received concomitant cisplatin-based chemotherapy. RESULTS: In 25 patients who presented with any component of treatment failure, the SUV was significantly higher than in the remaining patients without any such failure. Patients having tumors with high FDG uptake had a significantly lower 3-year local control (55% v 86%, P =.01) and DFS (42% v 79%, P =.005) compared with patients having low uptake tumors. In the multivariate analysis, the only factor that retained its significance for DFS was SUV category, whereas T category was of borderline significance. For local control, T category remained a significant factor, whereas a lower local control was observed for tumors with a high SUV compared with those with low SUV. CONCLUSION: FDG uptake, as measured by the SUV, has potential value in predicting local control and DFS in head and neck carcinomas treated by RT. High FDG uptake may be a useful parameter for identifying patients requiring more aggressive treatment approaches. PMID- 11870186 TI - Maximizing local control and organ preservation in stage IV squamous cell head and neck cancer With hyperfractionated radiation and concurrent chemotherapy. AB - PURPOSE: Results are reported from an aggressive chemoradiotherapy protocol for advanced squamous cell head and neck cancer. PATIENTS AND METHODS: Patients with advanced squamous cell head and neck cancer were treated with hyperfractionated radiation therapy (72 Gy at 1.2 Gy twice per day) and two courses of concurrent chemotherapy with fluorouracil (1,000 mg/m(2)/d) and cisplatin (20 mg/m(2)/d), both given as 96-hour continuous intravenous infusions during weeks 1 and 4 of radiation therapy. Primary-site resection was reserved for residual or recurrent primary-site disease after chemoradiotherapy. Neck dissection was considered for N2 or greater disease, irrespective of clinical response, and for residual or recurrent neck disease after nonoperative treatment. RESULTS: Forty-one patients with stage IV disease were treated. Toxicity was significant, with grade 3 to 4 mucositis in 98%, dysphagia in 88%, and skin reaction in 85%. Neutropenic fever requiring hospitalization occurred in 51%. Despite feeding tube placement in 35 patients (85%), the mean weight loss during chemoradiotherapy was 13.3% of initial body weight. One patient died during treatment as a result of a pulmonary embolus. At a median follow-up period of 30 months, the 3-year Kaplan-Meier projected overall survival was 59%, disease-specific survival 69%, likelihood of local control without surgical resection 91%, and local control with surgical resection 97%. The likelihood of distant disease control at 3 years was 74%, and distant metastases were present in eight of 13 patients who died. CONCLUSION: This chemoradiotherapy schedule produces considerable but manageable toxicity. Survival and organ preservation are excellent for this poor-prognosis patient cohort. Distant metastases are the most common cause of treatment failure. PMID- 11870187 TI - Skin lesions in melanoma and Kaposi's sarcoma. Case 1. Detection of circulating malignant cells by RT-PCR in a case of cutaneous melanoma in complete regression. PMID- 11870189 TI - Skin lesions in melanoma and Kaposi's sarcoma. Case 3. Familial classic Mediterranean Kaposi's sarcoma. PMID- 11870188 TI - Skin lesions in melanoma and Kaposi's sarcoma. Case 2. Dermoscopic features of metastases from cutaneous melanoma mimicking benign nevi and primary melanoma. PMID- 11870190 TI - Discussing hospice care. PMID- 11870191 TI - Combined chemotherapy trials require combined pharmacogenetic approaches. PMID- 11870192 TI - Impact of high-dose chemotherapy on peripheral T-cell lymphomas. PMID- 11870193 TI - Adjuvant radiation therapy after complete resection of non-small-cell lung cancer. PMID- 11870194 TI - Unexpected toxicity of combination thalidomide and interferon alpha-2a treatment in metastatic renal cell carcinoma. PMID- 11870197 TI - The management of tremor. PMID- 11870199 TI - Odd and unusual movement disorders. PMID- 11870198 TI - Diagnosis and management of atypical parkinsonian syndromes. PMID- 11870200 TI - Medical management of Parkinson's disease. PMID- 11870201 TI - Mind and movement: the neuropsychiatry of movement disorders. PMID- 11870202 TI - Surgery for movement disorders. PMID- 11870204 TI - My 9.11 reality call. By Caveman. PMID- 11870205 TI - The chromosome replication cycle. PMID- 11870206 TI - Repairing a torn cell surface: make way, lysosomes to the rescue. AB - Biological membranes are often described as 'self-sealing' structures. If indeed membranes do have an inherent capacity for repair, does this explain how a cell can rapidly reseal a very large (1-1000 microm(2)) disruption in its plasma membrane? It is becoming increasingly clear that, in nucleated animal cells, the cytoplasm plays an active and essential role in resealing. A rapid and apparently chaotic membrane fusion response is initiated locally in the cytoplasm by the Ca(2+) that floods in through a disruption: cytoplasmic vesicles are thereby joined with one another (homotypically) and with the surrounding plasma membrane (exocytotically). As a consequence, internal membrane is added to cell surface membrane at the disruption site. In the case of large disruptions, this addition is hypothesized to function as a 'patch'. In sea urchin eggs, the internal compartment used is the yolk granule. Several recent studies have significantly advanced our understanding of how cells survive disruption-inducing injuries. In fibroblasts, the lysosome has been identified as a key organelle in resealing. Protein markers of the lysosome membrane appear on the surface of fibroblasts at sites of disruption. Antibodies against lysosome-specific proteins, introduced into the living fibroblast, inhibit its resealing response. In gastric epithelial cells, local depolymerization of filamentous actin has been identified as a crucial step in resealing: it may function to remove a barrier to lysosome-plasma membrane contact leading to exocytotic fusion. Plasma membrane disruption in epithelial cells induces depolymerization of cortical filamentous actin and, if this depolymerization response is inhibited, resealing is blocked. In the Xenopus egg, the cortical cytoskeleton has been identified as an active participant in post-resealing repair of disruption-related damage to underlying cell cortex. A striking, highly localized actin polymerization response is observable around the margin of cortical defects. A myosin powered contraction occurring within this newly formed zone of F-actin then drives closure of the defect in a purse-string fashion. PMID- 11870207 TI - Twinfilin, a molecular mailman for actin monomers. AB - Twinfilin is a ubiquitous actin-monomer-binding protein that is composed of two ADF-homology domains. It forms a 1:1 complex with ADP-actin-monomers, inhibits nucleotide exchange on actin monomers and prevents assembly of the monomer into filaments. The two ADF-H domains in twinfilin probably have 3D structures similar to those of the ADF/cofilin proteins and overlapping actin-binding sites. Twinfilin also interacts with PtdIns(4,5)P(2), which inhibits its actin-monomer sequestering activity in vitro. Mutations in the twinfilin gene result in defects in the bipolar budding pattern in S. cerevisiae and in a rough eye phenotype and aberrant bristle morphology in Drosophila melanogaster. These phenotypes are caused by the uncontrolled polymerization of actin filaments in the absence of twinfilin. Studies on budding yeast suggest that twinfilin contributes to actin filament turnover by localizing actin monomers, in their 'inactive' ADP-form, to the sites of rapid filament assembly. This is mediated through direct interactions between twinfilin and capping protein. Therefore, twinfilin might serve as a link between rapid actin filament depolymerization and assembly in cells. PMID- 11870208 TI - F-actin ring formation and the role of F-actin cables in the fission yeast Schizosaccharomyces pombe. AB - Cells of the fission yeast Schizosaccharomyces pombe divide by the contraction of the F-actin ring formed at the medial region of the cell. We investigated the process of F-actin ring formation in detail using optical sectioning and three dimensional reconstruction fluorescence microscopy. In wild-type cells, formation of an aster-like structure composed of F-actin cables and accumulation of F-actin cables were recognized at the medial cortex of the cell during prophase to metaphase. The formation of the aster-like structure seemed to initiate from branching of the longitudinal F-actin cables at a site near the spindle pole bodies, which had been duplicated but not yet separated. A single cable extended from the aster and encircled the cell at the equator to form a primary F-actin ring during metaphase. During anaphase, the accumulated F-actin cables were linked to the primary F-actin ring, and then all of these structures seemed to be packed to form the F-actin ring. These observations suggest that formation of the aster-like structure and the accumulation of the F-actin cables at the medial region of the cell during metaphase may be required to initiate the F-actin ring formation. In the nda3 mutant, which has a mutation in ss-tubulin and has been thought to be arrested at prophase, an F-actin ring with accumulated F-actin cables similar to that of anaphase wild-type cells was formed at a restrictive temperature. Immediately after shifting to a permissive temperature, this structure changed into a tightly packed ring. This suggests that the F-actin ring formation progresses beyond prophase in the nda3 cells once the cells enter prophase. We further examined F-actin structures in both cdc12 and cdc15 early cytokinesis mutants. As a result, Cdc12 seemed to be required for the primary F actin ring formation during prophase, whereas Cdc15 may be involved in both packing the F-actin cables to form the F-actin ring and rearrangement of the F actin after anaphase. In spg1, cdc7 and sid2 septum initiation mutants, the F actin ring seemed to be formed in order. PMID- 11870210 TI - A requirement for the Abnormal Spindle protein to organise microtubules of the central spindle for cytokinesis in Drosophila. AB - Drosophila abnormal spindle (asp) mutants exhibit a mitotic metaphase checkpoint arrest with abnormal spindle poles, which reflects a requirement for Asp for the integrity of microtubule organising centres (MTOCs). In male meiosis, the absence of a strong spindle integrity checkpoint enables asp mutant cells to proceed through anaphase and telophase. However, the central spindle region is not correctly organised and cells frequently fail to complete cytokinesis. This contrasts with meiosis in wild-type males where at late anaphase a dense array of microtubules forms in the central spindle region that has Asp localised at its border. We speculate that Asp is associated with the minus ends of microtubules that have been released from the spindle poles to form the central spindle. A parallel situation arises in female meiosis where Asp not only associates with the minus ends of microtubules at the acentriolar poles but also with the central spindle pole body that forms between the two tandem spindles of meiosis II. Upon fertilisation, Asp is also recruited to the MTOC that nucleates the sperm aster. Asp is required for growth of the microtubules of the sperm aster, which in asp mutants remains diminutive and so prevents migration of the pronuclei. PMID- 11870209 TI - The small GTPase Rab22 interacts with EEA1 and controls endosomal membrane trafficking. AB - Rab22a is a small GTPase that is expressed ubiquitously in mammalian tissues and displays the highest sequence homology to Rab5. In BHK-21 cells, overexpression of the wild-type Rab22a caused formation of abnormally large vacuole-like structures containing the early-endosomal antigen EEA1 but not Rab11, a marker of recycling endosomes or the late-endosomal/lysosomal markers LAMP-1 and lyso-bis phosphatidic acid. In HeLa cells, overexpressed Rab22a was found on smaller EEA1 positive endosomes, but a portion of the protein was also found in the Golgi complex. Using the yeast two-hybrid system and a biochemical pull-down assay, the GTP-bound form of Rab22a was found to interact with the N-terminus of EEA1. In HeLa cells overexpressing Rab22a or its mutants affected in the GTPase cycle, no significant changes were observed in the uptake of Alexa-transferrin. However, the GTPase-deficient Rab22a Q64L mutant caused a redistribution of transferrin positive endosomes to the leading edges of cells and a fragmentation of the Golgi complex. In BHK cells, the Q64L mutant caused the accumulation of a fluid phase marker, TRITC-dextran, and a lysosomal hydrolase, aspartylglucosaminidase, in abnormal vacuole-like structures that contained both early and late endosome markers. Both the wild-type Rab22a and the Q64L mutant were found to interfere with the degradation of EGF. These results suggest that Rab22a may regulate the dynamic interactions of endosomal compartments and it may be involved in the communication between the biosynthetic and early endocytic pathways. PMID- 11870211 TI - The expression, lamin-dependent localization and RNAi depletion phenotype for emerin in C. elegans. AB - Emerin belongs to the LEM-domain family of nuclear membrane proteins, which are conserved in metazoans from C. elegans to humans. Loss of emerin in humans causes the X-linked form of Emery-Dreifuss muscular dystrophy (EDMD), but the disease mechanism is not understood. We have begun to address the function of emerin in C. elegans, a genetically tractable nematode. The emerin gene (emr-1) is conserved in C. elegans. We detect Ce-emerin protein in the nuclear envelopes of all cell types except sperm, and find that Ce-emerin co-immunoprecipitates with Ce-lamin from embryo lysates. We show for the first time in any organism that nuclear lamins are essential for the nuclear envelope localization of emerin during early development. We further show that four other types of nuclear envelope proteins, including fellow LEM-domain protein Ce-MAN1, as well as Ce lamin, UNC-84 and nucleoporins do not depend on Ce-emerin for their localization. This result suggests that emerin is not essential to organize or localize the only lamin (B-type) expressed in C. elegans. We also analyzed the RNAi phenotype resulting from the loss of emerin function in C. elegans under laboratory growth conditions, and found no detectable phenotype throughout development. We propose that C. elegans is an appropriate system in which to study the molecular mechanisms of emerin function in vivo. PMID- 11870212 TI - Kinesins klp5(+) and klp6(+) are required for normal chromosome movement in mitosis. AB - Proper mitotic chromosome segregation requires dynamic interactions between spindle microtubules and kinetochores. Here we demonstrate that two related fission yeast kinesins, klp5(+) and klp6(+), are required for normal chromosome segregation in mitosis. Null mutants frequently lack a normal metaphase chromosome alignment. Chromosome pairs move back and forth along the spindle for an extended period prior to sister chromatid separation, a phenotype reminiscent of the loss of CENP-E in metazoans. Ultimately, sister chromatids segregate, regardless of chromosome position along the spindle, and viable daughter cells are usually produced. The initiation of anaphase B is sometimes delayed, but the rate of spindle elongation is similar to wildtype. Despite a delay, anaphase B often begins before anaphase A is completed. The klp5Delta and klp6Delta null mutants are synthetically lethal with a deletion of the spindle assembly checkpoint gene, bub1(+), several mutants in components of the anaphase promoting complex, and a cold sensitive allele of the kinetochore and microtubule-binding protein, Dis1p. Klp5p-GFP and Klp6p-GFP localize to kinetochores from prophase to the onset of anaphase A, but relocalize to the spindle midzone during anaphase B. These data indicate that Klp5p and Klp6p are kinetochore kinesins required for normal chromosome movement in prometaphase. PMID- 11870213 TI - Perinuclear localization of huntingtin as a consequence of its binding to microtubules through an interaction with beta-tubulin: relevance to Huntington's disease. AB - Huntington's disease results from an expansion of a series of glutamine repeats in the protein huntingtin. We have discovered from immunopurification studies that huntingtin combines specifically with the beta subunit of tubulin. This binding explains why huntingtin can be shown on assembled microtubules by electron microscopy. Immunostaining shows that most of the huntingtin in the cytoplasm is associated with microtubules. Huntingtin is particularly abundant in the perinuclear region, where it is also associated with microtubules and in the centrosomal region, where it co-localizes with gamma-tubulin. In Huntington's disease, inclusions are often nuclear or perinuclear. Since the perinuclear concentration of huntingtin does not depend on the number of its glutamine repeats, we propose that inclusions are found in perinuclear and intranuclear locations because the beta-tubulin binding property of huntingtin brings it to the perinuclear region, from which it readily gains access to the nucleus. The mutational glutamine expansion then promotes insolubility and results in an inclusion. PMID- 11870214 TI - Phenotypic characterization of Drosophila ida mutants: defining the role of APC5 in cell cycle progression. AB - We have cloned and characterized the ida gene that is required for proliferation of imaginal disc cells during Drosophila development. IDA is homologous to APC5, a subunit of the anaphase-promoting complex (APC/cyclosome). ida mRNA is detected in most cell types throughout development, but it accumulates to its highest levels during early embryogenesis. A maternal component of IDA is required for the production of eggs and viable embryos. Homozygous ida mutants display mitotic defects: they die during prepupal development, lack all mature imaginal disc structures, and have abnormally small optic lobes. Cytological observations show that ida mutant brains have a high mitotic index and many imaginal cells contain an aneuploid number of aberrant overcondensed chromosomes. However, cells are not stalled in metaphase, as mitotic stages in which chromosomes are orientated at the equatorial plate are never observed. Interestingly, some APC/C-target substrates such as cyclin B are not degraded in ida mutants, whereas others controlling sister-chromatid separation appear to be turned over. Taken together, these results suggest a model in which IDA/APC5 controls regulatory subfunctions of the anaphase-promoting complex. PMID- 11870215 TI - The involvement of lipid rafts in the regulation of integrin function. AB - Integrin activity on cells such as T lymphocytes is tightly controlled. Here we demonstrate a key role for lipid rafts in regulating integrin function. Without stimulation integrin LFA-1 is excluded from lipid rafts, but following activation LFA-1 is mobilised to the lipid raft compartment. An LFA-1 construct from which the I domain has been deleted mimics activated integrin and is constitutively found in lipid rafts. This correlation between integrin activation and raft localisation extends to a second integrin, alpha4beta1, and the clustering of alpha4beta1 is also raft dependent. Both LFA-1 and alpha4beta1-mediated adhesion is dependent upon intact lipid rafts providing proof of the functional relevance of the lipid raft localisation. Finally we find that non-raft integrins are excluded from the rafts by cytoskeletal constraints. The presence of integrin in lipid rafts under stimulating conditions that activate these receptors strongly indicates that the rafts have a key role in positively regulating integrin activity. PMID- 11870216 TI - The CDK inhibitor NtKIS1a is involved in plant development, endoreduplication and restores normal development of cyclin D3; 1-overexpressing plants. AB - Plant development requires stringent controls between cell proliferation and cell differentiation. Proliferation is positively regulated by cyclin dependent kinases (CDKs). Acting in opposition to CDKs are CDK inhibitors (CKIs). The first tobacco CKI (NtKIS1a) identified was shown to inhibit in vitro the kinase activity of CDK/cyclin complexes and to interact with CDK and D-cyclins. However, these features, which are common to other plant and animal CKIs already characterised, did not provide information about the function of NtKIS1a in plants. Thus, to gain insight into the role of NtKIS1a and especially its involvement in cell proliferation during plant development, we generated transgenic Arabidopsis thaliana plants that overexpress NtKIS1a. These plants showed reduced growth with smaller organs that contained larger cells. Moreover, these plants displayed modifications in plant morphology. These results demonstrated that plant organ size and shape, as well as organ cell number and cell size, might be controlled by modulation of the single NtKIS1a gene activity. Since in mammals, D-cyclins control cell cycle progression in a CDK-dependent manner but also play a CDK independent role by sequestering the CKIs p27(Kip1) and p21(Cip1), we tested the significance of cyclin D-CKI interaction within a living plant. With this aim, NtKIS1a and AtCycD3;1 were overexpressed simultaneously in plants by two different methods. Our results demonstrated that overexpression of the CKI NtKIS1a restores essentially normal development in plants overexpressing AtCycD3;1, providing the first evidence of cyclin D-CKI co operation within the context of a living plant. PMID- 11870217 TI - Generation of diacylglycerol molecular species through the cell cycle: a role for 1-stearoyl, 2-arachidonyl glycerol in the activation of nuclear protein kinase C betaII at G2/M. AB - Protein kinase C (PKC) is a family of 11 isoenzymes that are differentially involved in the regulation of cell proliferation. PKC-betaII, a mitotic lamin kinase, has been shown previously to translocate to the nucleus at G(2)/M and this was coupled to the generation of nuclear diacylglycerol. However, it is not clear how isoenzyme selective translocation and nuclear targeting is achieved during cell cycle. To investigate further the role of nuclear diacylglycerol we measured PKC isoenzyme translocation and analysed diacylglycerol species at different stages of the cell cycle in U937 cells synchronized by centrifugal elutriation. Translocation of PKC-betaII to the membrane fraction, an indicator of activation, occurred at S and G(2)/M, although PKC-betaII was targeted to the nucleus only at G(2)/M. Levels of nuclear diacylglycerol, specifically tetraunsaturated species, increased during G(2)/M. By contrast, there were no obvious changes in nuclear phosphatidic acid species or mass. 1-stearoyl, 2 arachidonyl glycerol (SAG), the major polyunsaturated nuclear diacylglycerol, was able to activate classical PKC isoenzymes (PKC-alpha and beta), but was less effective for activation of novel isoenzymes (PKC-delta), in an in vitro PKC assay. We propose that PKC-betaII nuclear translocation during G(2)/M phase transition is mediated in part by generation of SAG at the nucleus. PMID- 11870218 TI - Human myosin-Vc is a novel class V myosin expressed in epithelial cells. AB - Class V myosins are one of the most ancient and widely distributed groups of the myosin superfamily and are hypothesized to function as motors for actin-dependent organelle transport. We report the discovery and initial characterization of a novel member of this family, human myosin-Vc (Myo5c). The Myo5c protein sequence shares approximately 50% overall identity with the two other class V myosins in vertebrates, myosin-Va (Myo5a) and myosin-Vb (Myo5b). Systematic analysis of the mRNA and protein distribution of these myosins indicates that Myo5a is most abundant in brain, whereas Myo5b and Myo5c are expressed chiefly in non-neuronal tissues. Myo5c is particularly abundant in epithelial and glandular tissues including pancreas, prostate, mammary, stomach, colon and lung. Immunolocalization in colon and exocrine pancreas indicates that Myo5c is expressed chiefly in epithelial cells. A dominant negative approach using a GFP Myo5c tail construct in HeLa cells reveals that the Myo5c tail selectively colocalizes with and perturbs a membrane compartment containing the transferrin receptor and rab8. Transferrin also accumulates in this compartment, suggesting that Myo5c is involved in transferrin trafficking. As a class V myosin of epithelial cells, Myo5c is likely to power actin-based membrane trafficking in many physiologically crucial tissues of the human body. PMID- 11870219 TI - Laminin assembles into separate basement membrane and fibrillar matrices in Schwann cells. AB - Laminins are important for Schwann cell basement membrane assembly and axonal function. In this study, we found that exogenous laminin-1, like neuromuscular laminins-2/4, formed two distinct extracellular matrices on Schwann cell surfaces, each facilitated by laminin polymerization. Assembly of one, a densely distributed reticular matrix, was accompanied by a redistribution of cell-surface dystroglycan and cytoskeletal utrophin into matrix-receptor-cytoskeletal complexes. The other, a fibrillar matrix, accumulated in separate zones associated with pre-existing beta1-integrin arrays. The laminin-1 fragment E3 (LG modules 4-5), which binds dystroglycan and heparin, inhibited reticular-matrix formation. By contrast, beta1-integrin blocking antibody (Ha2/5) prevented fibrillar assembly. Ultrastructural analysis revealed that laminin treatment induced the formation of a linear electron-dense extracellular matrix (lamina densa) separated from plasma membrane by a narrow lucent zone (lamina lucida). This structure was considerably reduced with non-polymerizing laminin, fully blocked by E3, and unaffected by Ha2/5. Although it formed in the absence of type IV collagen, it was nonetheless able to incorporate this collagen. Finally, cell competency to bind laminin and form a basement membrane was passage-dependent. We postulate that laminin induces the assembly of a basement membrane on competent cell surfaces probably mediated by anchorage through LG 4-5. Upon binding, laminin interacts with dystroglycan, mobilizes utrophin, and assembles a 'nascent' basement membrane, independent of integrin, that is completed by incorporation of type IV collagen. However, the fibrillar beta1-integrin dependent matrix is unlikely to be precursor to basement membrane. PMID- 11870220 TI - Disruption of microtubules uncouples budding and nuclear division in Toxoplasma gondii. AB - The tachyzoite stage of the protozoan parasite Toxoplasma gondii has two populations of microtubules: spindle microtubules and subpellicular microtubules. To determine how these two microtubule populations are regulated, we investigated microtubule behavior during the cell cycle following treatment with microtubule disrupting drugs. Previous work had established that the microtubule populations are individually nucleated by two distinct microtubule-organizing centers (MTOCs): the apical polar ring for the subpellicular microtubules and spindle pole plaques/centrioles for the spindle microtubules. When replicating tachyzoites were treated with 0.5 microM oryzalin or 1.0 mM colchicine they retained the capacity to form a spindle and undergo nuclear division. Although these parasites could complete budding, they lost the bulk of their subpellicular microtubules and the ability to reinvade host cells. Both nascent spindle and subpellicular microtubules were disrupted in 2.5 microM oryzalin or 5.0 mM colchicine. Under these conditions, parasites grew in size and replicated their genome but were incapable of nuclear division. After removal from 0.5 microM oryzalin, Toxoplasma tachyzoites were able to restore normal subpellicular microtubules and a fully invasive phenotype. When oryzalin was removed from Toxoplasma tachyzoites treated with 2.5 microM drug, the parasites attempted to bud as crescent-shaped tachyzoites. Because the polyploid nuclear mass could not be correctly segregated, many daughter parasites lacked nuclei altogether although budding and scission from the maternal mass was able to be completed. Multiple MTOCs permit Toxoplasma tachyzoites to control nuclear division independently from cell polarity and cytokinesis. This unusual situation grants greater cell cycle flexibility to these parasites but abolishes the checks for coregulation of nuclear division and cytokinesis found in other eukaryotes. PMID- 11870221 TI - The editosome for cytidine to uridine mRNA editing has a native complexity of 27S: identification of intracellular domains containing active and inactive editing factors. AB - Apolipoprotein B mRNA cytidine to uridine editing requires the assembly of a multiprotein editosome comprised minimally of the catalytic subunit, apolipoprotein B mRNA editing catalytic subunit 1 (APOBEC-1), and an RNA-binding protein, APOBEC-1 complementation factor (ACF). A rat homolog has been cloned with 93.5% identity to human ACF (huACF). Peptide-specific antibodies prepared against huACF immunoprecipitated a rat protein of similar mass as huACF bound to apolipoprotein B (apoB) RNA in UV cross-linking reactions, thereby providing evidence that the p66, mooring sequence-selective, RNA-binding protein identified previously in rat liver by UV cross-linking and implicated in editosome assembly is a functional homolog of huACF. The rat protein (p66/ACF) was distributed in both the nucleus and cytoplasm of rat primary hepatocytes. Within a thin section, a significant amount of total cellular p66/ACF was cytoplasmic, with a concentration at the outer surface of the endoplasmic reticulum. Native APOBEC-1 co-fractionated with p66/ACF in the cytoplasm as 60S complexes. In the nucleus, the biological site of apoB mRNA editing, native p66/ACF, was localized to heterochromatin and fractionated with APOBEC-1 as 27S editosomes. When apoB mRNA editing was stimulated in rat primary hepatocytes with ethanol or insulin, the abundance of p66/ACF in the nucleus markedly increased. It is proposed that the heterogeneity in size of complexes containing editing factors is functionally significant and reflects functionally engaged editosomes in the nucleus and an inactive cytoplasmic pool of factors. PMID- 11870222 TI - Substrate proteolysis is inhibited by dominant-negative Nedd4 and Rsp5 mutants harboring alterations in WW domain 1. AB - Mammalian Nedd4 and its budding yeast orthologue Rsp5 are members of a large family of HECT-domain-containing ubiquitin ligases. Besides possessing a Ca(2+)/lipid-binding domain, both ligases have multiple protein-interacting modules termed WW domains. The C-terminal WW domains mediate interactions with substrates, but the function of the first WW domain remains unclear. We found that expression of a WW domain 1 Nedd4 mutant inhibits the growth of budding yeast by affecting the rsp5-ole1 pathway. The WW domain 1 mutant-induced phenotype is suppressed by ole1 cDNA overexpression or oleic acid supplementation of growth media and ole1 RNA levels are reduced in cells expressing this Nedd4 mutant. Also, the WW domain 1 Nedd4 mutant associates via WW domains 2 and 3 with Spt23, a Rsp5 target and ole1 transactivator. The dominant-negative activity of this mutant is associated with promoting accumulation of unprocessed Spt23 and inhibiting generation of processed and presumably active protein. Also, Spt23 processing is inhibited by a Nedd4 mutant that lacks ubiquitin ligase activity and Spt23-binding-competent Rsp5 mutants harboring WW domain 1 or ligase domain mutations. Interestingly, in mammalian cells, wild-type Nedd4 promotes proteasome mediated degradation of the precursor form of Spt23. WW domain 1 and ligase domain Nedd4 mutants block its degradation. These results indicate that WW domain 1 of these ligases interacts with cofactors that are required for ubiquitin/proteasome-dependent proteolysis of bound substrates. PMID- 11870223 TI - Cell-type-specific and selectively induced expression of members of the p24 family of putative cargo receptors. AB - Members of the p24 family of type I transmembrane proteins are highly abundant in transport vesicles and are thought to be involved in selective protein transport between the endoplasmic reticulum and the Golgi complex. The p24 proteins have been grouped into four subfamilies (alpha, beta, gamma, and delta) and appear to assemble into tetrameric complexes that contain only one representative from each subfamily. Here we molecularly dissected the p24 family in a single cell type, namely in the intermediate pituitary melanotrope cells of the amphibian Xenopus laevis. The biosynthetic activity of these cells for production of their major cargo protein proopiomelanocortin (POMC) can be physiologically manipulated via the process of background adaptation (similar30-fold induction, with highly active cells in black toads and virtually inactive cells in white animals). Extensive cDNA library screening revealed the identity of six p24 proteins expressed in the Xenopus melanotrope cells, namely one member of the p24alpha (alpha(3)), one of the p24beta (beta(1)), two of the p24gamma (gamma(2), gamma(3)) and two of the p24delta (delta(1), delta(2)) subfamily. Two other Xenopus p24 proteins, Xp24alpha(2) and -gamma(1), were not expressed in the melanotrope cells, pointing to cell-type specific p24 expression. Of the six melanotrope p24 proteins, the expression of four (Xp24alpha(3), -beta(1), gamma(3) and -delta(2)) was 20- to 30-fold induced in active versus inactive melanotropes, whereas that of the other two members (Xp24gamma(2) and -delta(1)) had not or only slightly increased. The four proteins were induced only in the intermediate melanotrope cells and not in the anterior pituitary cells, and displayed similar overall tissue distributions that differed from those of Xp24gamma(1), -gamma(2) and -delta(1). Together, our results reveal that p24 expression can be cell-type specific and selectively induced, and suggest that in Xenopus melanotrope cells an alpha(3)/beta(1)/gamma(3)/delta(2) p24 complex is involved in POMC transport through the early stages of the secretory pathway. PMID- 11870224 TI - Ephrin-A5 induces rounding, blebbing and de-adhesion of EphA3-expressing 293T and melanoma cells by CrkII and Rho-mediated signalling. AB - Eph receptor tyrosine kinases and ephrins regulate morphogenesis in the developing embryo where they effect adhesion and motility of interacting cells. Although scarcely expressed in adult tissues, Eph receptors and ephrins are overexpressed in a range of tumours. In malignant melanoma, increased Eph and ephrin expression levels correlate with metastatic progression. We have examined cellular and biochemical responses of EphA3-expressing melanoma cell lines and human epithelial kidney 293T cells to stimulation with polymeric ephrin-A5 in solution and with surfaces of defined ephrin-A5 densities. Within minutes, rapid reorganisation of the actin and myosin cytoskeleton occurs through activation of RhoA, leading to the retraction of cellular protrusions, membrane blebbing and detachment, but not apoptosis. These responses are inhibited by monomeric ephrin A5, showing that receptor clustering is required for this EphA3 response. Furthermore, the adapter CrkII, which associates with tyrosine-phosphorylated EphA3 in vitro, is recruited in vivo to ephrin-A5-stimulated EphA3. Expression of an SH3-domain mutated CrkII ablates cell rounding, blebbing and detachment. Our results suggest that recruitment of CrkII and activation of Rho signalling are responsible for EphA3-mediated cell rounding, blebbing and de-adhesion, and that ephrin-A5-mediated receptor clustering and EphA3 tyrosine kinase activity are essential for this response. PMID- 11870225 TI - Forskolin-mediated G1 arrest in acute lymphoblastic leukaemia cells: phosphorylated pRB sequesters E2Fs. AB - Increased intracellular levels of cAMP, induced by forskolin, lead to permanent G1 arrest of Reh cells. As expected, we observed a rapid dephosphorylation of the retinoblastoma protein (pRB) within 2 hours of forskolin treatment concomitant with reduced activity of the pRB-specific kinases. Interestingly, however, the dephosphorylation of pRB, as well as the inhibition of the kinase activities, was only transient, despite the permanent arrest of cells in G1. Importantly, although the pRB-specific kinases were fully active after 48 hours, pRB became only partially rephosphorylated. The transient dephosphorylation of pRB could be explained by the transient decrease in the activities of the pRB-specific kinases, but to understand why pRB became only partially rephosphorylated, despite fully activated kinases, we postulated that cAMP could activate a pRB directed phosphatase. It was therefore interesting to find that the phosphatase inhibitor, tautomycin, was able to abolish the forskolin-mediated dephosphorylation of pRB, without increasing the activities of the pRB-specific kinases. To understand how Reh cells expressing hyperphosphorylated forms of pRB can remain arrested in G1, we used three different methods to test for the ability of pRB to form functional complexes with the family of E2F transcription factors. As expected, we observed an increased complex formation between E2F-1, E2F-4 and pRB after 2 hours when pRB was in its most dephosphorylated state. Surprisingly, however, prolonged treatment with forskolin, which induced partial rephosphorylation of pRB, in fact further increased the complex formation between the E2Fs and pRB, and this also resulted in reduced E2F-promoter activity in vivo. These data imply that in Reh cells, partially phosphorylated forms of pRB retain the ability to inhibit E2F-promoter activity, and thereby prevent cells from entering into S-phase. PMID- 11870226 TI - Katanin inhibition prevents the redistribution of gamma-tubulin at mitosis. AB - Katanin is a microtubule-severing protein that is concentrated at mitotic spindle poles but katanin's function in the mitotic spindle has not been previously reported. Inhibition of katanin with either of two dominant-negative proteins or a subunit-specific antibody prevented the redistribution of gamma-tubulin from the centrosome to the spindle in prometaphase CV-1 cells as assayed by immunofluorescence microscopy. Because gamma-tubulin complexes can bind to pre existing microtubule minus ends, these results could be explained by a model in which the broad distribution of gamma-tubulin in the mitotic spindle is in part due to cytosolic gamma-tubulin ring complexes binding to microtubule minus ends generated by katanin-mediated microtubule severing. Because microtubules depolymerize at their ends, we hypothesized that a greater number of microtubule ends generated by severing in the spindle would result in an increased rate of spindle disassembly when polymerization is blocked with nocodazole. Indeed, katanin inhibition slowed the rate of spindle microtubule disassembly in the presence of nocodazole. However, katanin inhibition did not affect the rate of exchange between polymerized and unpolymerized tubulin as assayed by fluorescence recovery after photobleaching. These results support a model in which katanin activity regulates the number of microtubule ends in the spindle. PMID- 11870227 TI - A novel Ala(-3)Thr mutation in the signal peptide of human luteinizing hormone beta-subunit: potentiation of the inositol phosphate signalling pathway and attenuation of the adenylate cyclase pathway by recombinant variant hormone. AB - Upon screening for polymorphisms in the human luteinizing hormone beta-subunit (LH beta) gene, we discovered a novel mutation in the LH beta signal peptide with functional consequences for signal transduction in mouse Leydig tumour cells (mLTC-1). This G(52)A point mutation in exon 2 of the LH beta gene, detected in heterozygous form in several normal DNA samples, caused an Ala(-3)Thr amino acid substitution. Recombinant forms of wild-type (WT) and Ala(-3)Thr variant (V) LH were produced in human embryonic kidney (HEK) 293 cells and purified. The immunoreactivities of the recombinant LH were determined by immunofluorometric assays and in-vitro bioactivities in mLTC-1 cells were assessed by using cAMP, progesterone and inositol trisphosphate (IP(3)), and activation of mitogen activated protein kinase (MAPK) as end-points. Whereas both LH forms stimulated progesterone production and MAPK in similar fashion, WT-LH was more potent in stimulating cAMP, and V-LH was more potent in stimulating IP(3) generation. Both LH forms bound to LH receptors with similar affinities. No evidence was found for influence of the signal peptide mutation on efficacy of alpha- and beta-subunit dimerization. Sequencing of the recombinant V-LH beta protein also revealed that the mutation did not interfere with signal peptide cleavage. In summary, the present findings indicate that the Ala(-3)Thr mutation in the LH beta-subunit signal peptide has functional consequences, in the form of dissociation of stimulatory potency for different signal transduction pathways in vitro. PMID- 11870228 TI - Fas expression correlates with human germ cell degeneration in meiotic and post meiotic arrest of spermatogenesis. AB - Degeneration of human male germ cells was analysed by means of light (LM) and transmission electron (TEM) microscopy. The frequency of degenerating cells was correlated with that of Fas-expressing germ cells in human testes with normal spermatogenesis (n = 10), complete early maturation arrest (EMA) (n = 10) or incomplete late maturation arrest (LMA; n = 10) of spermatogenesis. LM analysis of testis sections with normal spermatogenesis indicated that degenerating germ cells were localized in the adluminal compartment of the seminiferous epithelium. TEM showed that apoptotic cells were mostly primary spermatocytes and, to a lesser extent, round or early elongating spermatids. Apoptotic germ cells appeared to be eliminated either in the seminiferous lumen or by Sertoli cell phagocytosis. An increased number of degenerating cells was observed in testes with LMA as compared with normal testes and testes with EMA of spermatogenesis (P < 0.001, Wilcoxon's rank sum test). Comparison of these results with those obtained from immunohistochemistry experiments demonstrated a tight correlation between the number of apoptotic cells and the number of Fas-expressing germ cells (P = 0.001, Spearman's rank = 0.69). These findings suggest that altered meiotic and post-meiotic germ cell maturation might be associated with an up-regulation of Fas gene expression capable of triggering apoptotic elimination of defective germ cells. PMID- 11870229 TI - Purification and characterization of human seminal plasma alpha-L-fucosidase. AB - Human seminal plasma alpha-L-fucosidase (EC 3.2.1.51) has been purified 7100-fold to very high purity and specific activity (83,000 nmol/min/mg protein) by affinity chromatography on agarose-epsilon-aminocaproyl-fucopyranosylamine. The purified alpha-L-fucosidase appeared to contain a single subunit of 56-57 kDa (as determined by SDS-PAGE and Western analysis). Lectin blotting and N-glycanase treatment studies indicated that this subunit is N-glycosylated and contains sialic acid residues. Human seminal plasma alpha-L-fucosidase was shown to contain three multimeric forms of 110, 236 and 314 kDa respectively (as determined by Sephadex G-200 chromatography) and therefore probably exists in dimeric, tetrameric and hexameric forms. Kinetic analysis with the 4 methylumbelliferyl-alpha-L-fucopyranoside (4MU-Fuc) substrate indicated a broad acidic optimum (pH 4.0-4.5) with a second neutral optimum (pH 6.4-7.4) with 60 80% of maximal activity. Apparent K(M) and V(max) values for the 4MU-Fuc substrate were determined to be 0.06 mmol/l and 92 micromol/min/mg protein respectively, using Lineweaver-Burk double reciprocal plots. Isoelectric focusing and neuraminidase treatment studies provided further evidence that the purified seminal plasma alpha-L-fucosidase is a sialoglycoprotein with several isoforms between pI values 5-7. The acidic isoforms between pI values 5-6 appear to be related chemically to the more neutral isoforms by sialic acid residues since neuraminidase treatment converted the former into the latter isoforms. PMID- 11870230 TI - Cell death and its suppression in human ovarian tissue culture. AB - In women with premature ovarian failure, fertility may be preserved by ovarian tissue culture in vitro. However, techniques for tissue culture and follicle maturation have remained suboptimal. Our aim was to characterize ovarian tissue degeneration in cultures and to establish a model for cell death research in cultured ovarian tissue. Precise knowledge on the process resulting in cell death in cultured ovarian tissue will ultimately facilitate work aimed at improving long-term culture conditions. Ovarian tissue apoptosis was studied in a serum free culture model in which nuclear DNA fragmentation was shown to occur within 24 h of the start of the culture. Activation of caspase-3 was detected in some stromal cells and a few oocytes. Since not all of the tissue exhibited signs of apoptosis and since DNA fragmentation increased over time, the tissue probably gradually dies by apoptosis. The antioxidant N-acetyl-L-cysteine (NAC; 25, 50 and 100 mmol/l) was found to inhibit this apoptosis. Thus, apoptosis appears to play a critical role in the degeneration of human ovarian cortical tissue cultures, and this cell death can be suppressed by NAC. The present tissue culture model can be used for identifying components capable of inhibiting cell death in vitro. PMID- 11870231 TI - Expression of cell adhesion molecules during human preimplantation embryo development. AB - Formation of a fully differentiated, implantation competent blastocyst requires the expression of a complex repertoire of molecules. However, the events that drive morphogenesis are poorly elucidated in the human embryo. In this work, we describe the amplification of representative cDNAs from morphologically and developmentally normal, individual human embryos at all stages from pronucleate to blastocyst. These cDNAs were probed to reveal the temporal expression pattern of cell adhesion molecules thought to play a key role in murine preimplantation embryo development. We demonstrated constitutive expression of beta actin, beta 1 and alpha 6 integrins, ZO-1 and E-cadherin, as shown previously in mouse embryos. No expression of beta 3, alpha 2, alpha 3 or alpha 7 integrins nor of L or P selectin was detected at any stage of preimplantation development. beta 5 integrin showed a regulated pattern of expression and was not expressed in blastocysts, while desmocollin-2 could only be detected at the blastocyst stage. Expression and localization of beta 1, beta 5 and alpha 6 integrins and ZO-1 and E-cadherin proteins was confirmed in blastocyst stage embryos by immunocytochemistry. We have identified differences in the expression of integrin molecules between mouse and human embryos, and propose a role for alpha v beta 5 and alpha 6 beta 1 integrin dimers in the human embryo at implantation. PMID- 11870232 TI - Identification of genes with higher expression in human uterine leiomyomas than in the corresponding myometrium. AB - We used a PCR-based subtraction method, representational difference analysis of cDNA (cDNA--RDA), to identify genes with a higher expression in leiomyomas in comparison with the corresponding myometrium during the proliferative phase of the menstrual cycle. Increased expression of the genes for pregnancy-associated plasma protein A (PAPPA), tomoregulin, cellular retinoid acid binding protein 1 (CRABP1), zinc finger protein 185 (ZFP 185) and latent transforming growth factor beta binding protein 2 (LTBP2) was demonstrated in individual leiomyoma samples compared with corresponding myometrium. Additionally, a specific positive immunostaining of LTBP2 was found in the smooth muscle cells of both leiomyomas and myometrium. These genes may be part of previously unidentified molecular mechanisms responsible for the selective growth advantage of leiomyomas compared with myometrium. This work expands our knowledge about the molecular nature of leiomyomas and provides novel candidate genes to further explore in relation to their function during leiomyoma growth. PMID- 11870233 TI - Th1- and Th2-like cytokine production by first trimester decidual large granular lymphocytes is influenced by HLA-G and HLA-E. AB - During normal early pregnancy, a particular immune environment in the decidua and the expression of non-classical HLA-G and HLA-E molecules on the invading trophoblast are assumed to be essential for the tolerance of the fetus. To assess whether HLA-G and HLA-E influence the cytokine production of their putative target cells [large granular lymphocytes (LGL)], we analysed the concentrations of tumour necrosis factor (TNF-alpha), interferon (IFN)-gamma, interleukin (IL) 10, IL-13 and granulocyte-macrophage colony stimulating factor (GM-CSF) in supernatants of isolated first trimester LGL co-cultured with HLA-G or HLA-E transfected K-562 leukaemia cells lacking the classical HLA class I and II molecules. In comparison with that observed with untransfected K-562 cells, co culture of LGL with HLA-G-expressing cells significantly reduced the concentration of all cytokines investigated (TNF-alpha, IL-10 and GM-CSF, P < 0.01; IFN-gamma and IL-13, P < 0.05). In contrast, co-culture of LGL with HLA-E expressing cells significantly (P < 0.01) decreased only IL-10 production, although a strong tendency towards reduced IL-13 levels was also observed. In the co-culture system presented, membrane-bound HLA-G and, to a lesser extent, HLA-E expression affected cytokine release by decidual LGL in a manner not consistent with the Th1/Th2 paradigm. In conclusion, our data are indicative of a general immune-suppressive effect of HLA-G on LGL activity. PMID- 11870234 TI - Control of the human inhibin alpha chain promoter in cytotrophoblast cells differentiating into syncytium. AB - Inhibins are dimeric proteins consisting of a common alpha subunit linked to one of the beta subunits, beta A or beta B. During pregnancy, the placenta is the main source of inhibin A production and the in-vitro transformation of cytotrophoblast cells into syncytium is associated with an inhibin alpha subunit mRNA up-regulation. In this study, the 5' region of the human inhibin alpha gene was isolated and sequenced. Three transcription initiation sites were identified. When transiently transfected in trophoblast cells with a luciferase reporter vector, the sequence displayed promoter activity. DNase I footprinting and electrophoretic mobility shift assay (EMSA) analysis showed a specific DNA protein interaction in the promoter when using cytotrophoblast nuclear proteins. This interaction was weaker with syncytiotrophoblast nuclear proteins. Moreover, the deletion of this DNA-protein interaction region suppressed the promoter activity. In an attempt to identify this factor, the potential binding of known factors delta EF1, AP1 and NFE2 were excluded by competition EMSA experiments. We suggest that it may correspond to an undescribed protein interaction. The identification of the human inhibin alpha promoter could help in understanding the mechanisms modulating inhibin gene transcription. Moreover, the identification of a factor, whose presence is related to the trophoblast cell differentiation state, could help in understanding the transformation of cytotrophoblast cells into syncytium. PMID- 11870235 TI - Chemical stabilization of tetrahydrobiopterin by L-ascorbic acid: contribution to placental endothelial nitric oxide synthase activity. AB - The aim of this study was to characterize the mechanism of the chemical interaction between L-ascorbic acid (ASC) and tetrahydrobiopterin (BH(4)) in vitro and to examine its effect on the activity of endothelial nitric oxide synthase (eNOS) in first trimester human placentae. At room temperature, in Tris HCl buffer (pH 7.4), both ASC and BH(4) were readily oxidized by dissolved O(2) or H(2)O(2). BH(4) was more sensitive to auto-oxidation, while ASC was more susceptible to oxidation by H(2)O(2). Addition of 36 micromol/l BH(4) to 143 micromol/l ASC increased the initial rate of ASC oxidation 3.2-fold in a catalase sensitive manner, indicating that enhanced ASC oxidation is partly due to the formation of H(2)O(2). In the presence of catalase, BH(4) still stimulated 1.9 fold the initial rate of ASC oxidation, suggesting that another auto-oxidation product of BH(4), most probably quininoid-BH(2) (qBH(2)), could also stimulate ASC oxidation while itself being reduced back to BH(4). ASC prevented the auto oxidation of BH(4) in a concentration-dependent fashion, with 3 mmol/l ASC providing an almost complete stabilization of 25 micromol/l BH(4). Importantly, basal eNOS activity in placental microsomes was stimulated 2.5-fold by 0.5 micromol/l BH(4), and 0.5 mmol/l ASC enhanced the BH(4)-stimulation 1.4-fold, with a smaller effect on basal eNOS activity. Taken together, the findings support the notion that the stabilizing action of ASC on BH(4) is related to the ASC-mediated reductive reversal of the auto-oxidation process of BH(4). Moreover, we demonstrated that concentrations of ASC present in the placenta as a common vitamin C supply are sufficient to protect cellular free BH(4) and may contribute to the stimulation of placental eNOS activity. PMID- 11870236 TI - The effects of a cytokine suppressive anti-inflammatory drug on the output of prostaglandin E(2) and interleukin-1 beta from human fetal membranes. AB - Fetal membranes are a primary source of prostaglandins and pro-inflammatory cytokines implicated in human parturition, so the inhibition of inflammatory pathways may be of benefit in pregnancies complicated by premature labour. We have therefore investigated the effects of a cytokine-suppressant anti inflammatory drug (CSAID) on the output of prostaglandin E(2) (PGE(2)) and interleukin (IL)-1 beta from human fetal membranes in vitro. Bacterial endotoxin increased the expression of mRNA for IL-1 beta and type-2 cyclo-oxygenase (COX 2), and there were corresponding increases in the output of IL-1 beta protein and PGE(2). The CSAID decreased IL-1 beta protein, COX-2 expression and PGE(2) output, but not mRNA for IL-1 beta, indicating a post-translational effect on the production of IL-1 beta and a transcriptional affect on COX-2, with an overall reduction in PGE(2). These findings are consistent with the effects of CSAIDs in other systems, and indicate that they are of possible use in premature labour. PMID- 11870238 TI - Lack of association between polymorphisms in the testis-specific angiotensin converting enzyme gene and male infertility in an Asian population. AB - Angiotensin converting enzyme (ACE) is a membrane-bound dipeptidyl carboxy peptidase that generates vasoconstricting angiotensin II and inactivates vasodilating bradykinin. The ACE gene encodes two isozymes: the somatic isozyme (sACE) is found in many tissues including vascular endothelial cells, whereas the testis-specific isozyme (tACE) is expressed exclusively in developing spermatids and mature sperm. Thus, ACE might have physiological functions in addition to blood pressure regulation. Male mice lacking tACE activity show reduced fertility, indicating its importance in male fertility. In this study, we screened five recently defined tACE gene polymorphisms in 90 Singapore Chinese men with infertility and 84 fertile controls using PCR-based restriction fragment length polymorphism and DNA sequencing. However, only one of these polymorphisms was identified in both patient and control groups, the frequency of which was not significantly different in patients and controls. Thus, these ACE gene polymorphisms are unlikely to contribute to the pathogenesis of male infertility in the Singapore Chinese population. PMID- 11870237 TI - High frequency of DAZ1/DAZ2 gene deletions in patients with severe oligozoospermia. AB - Deletions of the DAZ gene family in distal Yq11 are always associated with deletions of the azoospermia factor c (AZFc) region, which we now estimate extends to 4.94 Mb. Because more Y gene families are located in this chromosomal region, and are expressed like the DAZ gene family only in the male germ line, the testicular pathology associated with complete AZFc deletions cannot predict the functional contribution of the DAZ gene family to human spermatogenesis. We therefore established a DAZ gene copy specific deletion analysis based on the DAZ BAC sequences in GenBank. It includes the deletion analysis of eight DAZ-DNA PCR markers [six DAZ-single nucleotide varients (SNVs) and two DAZ-sequence tag sites (STS)] selected from the 5' to the 3'end of each DAZ gene and a deletion analysis of the gene copy specific EcoRV and TaqI restriction fragments identified in the internal repetitive DAZ gene regions (DYS1 locus). With these diagnostic tools, 63 DNA samples from men with idiopathic oligozoospermia and 107 DNA samples from men with proven fertility were analysed for the presence of the complete DAZ gene locus, encompassing the four DAZ gene copies. In five oligozoospermic patients, we found a DAZ-SNV/STS and DYS1/EcoRV and TaqI fragment deletion pattern indicative for deletion of the DAZ1 and DAZ2 gene copies; one of these deletions could be identified as a 'de-novo' deletion because it was absent in the DAZ locus of the patient's father. The same DAZ deletions were not found in any of the 107 fertile control samples. We therefore conclude that the deletion of the DAZ1/DAZ2 gene doublet in five out of our 63 oligozoospermic patients (8%) is responsible for the patients' reduced sperm numbers. It is most likely caused by intrachromosomal recombination events between two long repetitive sequence blocks (AZFc-Rep1) flanking the DAZ gene structures. PMID- 11870239 TI - Pregnancy following preimplantation genetic diagnosis for Crouzon syndrome. AB - Crouzon syndrome is a dominantly inherited craniosynostosis syndrome which is caused by mutations in the fibroblast growth factor receptor 2 gene (FGFR2). However, a specific point mutation in the FGFR3 gene has also been shown to result in Crouzon syndrome associated with acanthosis nigricans. We report here the first method for preimplantation genetic diagnosis (PGD) of Crouzon syndrome based on multiplex PCR amplification followed by the direct detection of the causative mutation by single-stranded conformational polymorphism (SSCP) analysis. A highly polymorphic short tandem repeat (STR) locus was simultaneously analysed as a control against some forms of contamination. The mutation, carried by the female partner, was a de-novo substitution at codon 338 of the FGFR2 gene. The couple were found to be informative at the D21S11 STR locus. Two clinical PGD cycles were performed, resulting in the biopsy of 36 blastomeres, 25 of which showed amplification at the FGFR2 locus. All of the cells showed expected genotypes at the D21S11 locus with only one incidence of allele drop-out. A total of five embryos were transferred, two in the first cycle and three in the second, resulting in a singleton pregnancy. PMID- 11870240 TI - Bisphosphonates and osteoporosis. PMID- 11870241 TI - Hematologic and cytogenetic responses to imatinib mesylate in chronic myelogenous leukemia. AB - BACKGROUND: Chronic myelogenous leukemia (CML) is caused by the BCR-ABL tyrosine kinase, the product of the Philadelphia chromosome. Imatinib mesylate, formerly STI571, is a selective inhibitor of this kinase. METHODS: A total of 532 patients with late--chronic-phase CML in whom previous therapy with interferon alfa had failed were treated with 400 mg of oral imatinib daily. Patients were evaluated for cytogenetic and hematologic responses. Time to progression, survival, and toxic effects were also evaluated. RESULTS: Imatinib induced major cytogenetic responses in 60 percent of the 454 patients with confirmed chronic-phase CML and complete hematologic responses in 95 percent. After a median follow-up of 18 months, CML had not progressed to the accelerated or blast phases in an estimated 89 percent of patients, and 95 percent of the patients were alive. Grade 3 or 4 nonhematologic toxic effects were infrequent, and hematologic toxic effects were manageable. Only 2 percent of patients discontinued treatment because of drug related adverse events, and no treatment-related deaths occurred. CONCLUSIONS: Imatinib induced high rates of cytogenetic and hematologic responses in patients with chronic-phase CML in whom previous interferon therapy had failed. PMID- 11870242 TI - Intravenous zoledronic acid in postmenopausal women with low bone mineral density. AB - BACKGROUND: Bisphosphonates are effective agents for the management of osteoporosis. Their low bioavailability and low potency necessitate frequent administration on an empty stomach, which may reduce compliance. Gastrointestinal intolerance limits maximal dosing. Although intermittent intravenous treatments have been used, the optimal doses and dosing interval have not been systematically explored. METHODS: We studied the effects of five regimens of zoledronic acid, the most potent bisphosphonate, on bone turnover and density in 351 postmenopausal women with low bone mineral density in a one-year, randomized, double-blind, placebo-controlled trial. Women received placebo or intravenous zoledronic acid in doses of 0.25 mg, 0.5 mg, or 1 mg at three-month intervals. In addition, one group received a total annual dose of 4 mg as a single dose, and another received two doses of 2 mg each, six months apart. Lumbar-spine bone mineral density was the primary end point. RESULTS: There were similar increases in bone mineral density in all the zoledronic acid groups to values for the spine that were 4.3 to 5.1 percent higher than those in the placebo group (P<0.001) and values for the femoral neck that were 3.1 to 3.5 percent higher than those in the placebo group (P<0.001). Biochemical markers of bone resorption were significantly suppressed throughout the study in all zoledronic acid groups. Myalgia and pyrexia occurred more commonly in the zoledronic acid groups, but treatment-related dropout rates were similar to that in the placebo group. CONCLUSIONS: Zoledronic acid infusions given at intervals of up to one year produce effects on bone turnover and bone density as great as those achieved with daily oral dosing with bisphosphonates with proven efficacy against fractures, suggesting that an annual infusion of zoledronic acid might be an effective treatment for postmenopausal osteoporosis. PMID- 11870243 TI - The prevalence of proteolytic antibodies against factor VIII in hemophilia A. AB - BACKGROUND: Factor VIII inhibitors are IgG alloantibodies that arise during replacement therapy in 25 to 50 percent of patients with severe hemophilia A. The hydrolysis of factor VIII by anti--factor VIII antibodies has been proposed as a mechanism of inactivation of factor VIII. METHODS: We purified IgG from patients with severe hemophilia A. The proteolytic activity of the antibodies was assessed by incubating the IgG with biotinylated human factor VIII and analyzing patterns of factor VIII cleavage by sodium dodecyl sulfate--polyacrylamide-gel electrophoresis and immunoblotting. The controls were normal human IgG and IgG purified from plasma of patients with hemophilia who did not have inhibitory antibodies. RESULTS: Significant proteolytic activity was detected in IgG from 13 of 24 inhibitor-positive patients. No hydrolytic activity was detected in control antibodies of IgG from patients without inhibitors. The rate of hydrolysis of factor VIII by purified IgG correlated positively with the factor VIII- neutralizing activity of IgG in plasma (r2=0.67, P=0.029). Principal-component analysis of migration profiles of digestion fragments demonstrated the heterogeneity of the catalytic potential of factor VIII inhibitors among patients. CONCLUSIONS: Proteolysis is a mechanism by which IgG antibodies against factor VIII can inactivate factor VIII. PMID- 11870244 TI - An outbreak of eosinophilic meningitis caused by Angiostrongylus cantonensis in travelers returning from the Caribbean. AB - BACKGROUND: Outbreaks of eosinophilic meningitis caused by the roundworm Angiostrongylus cantonensis are rarely reported, even in regions of endemic infection such as Southeast Asia and the Pacific Basin. We report an outbreak of A. cantonensis meningitis among travelers returning from the Caribbean. METHODS: We conducted a retrospective cohort study among 23 young adults who had traveled to Jamaica. We used a clinical definition of eosinophilic meningitis that included headache that began within 35 days after the trip plus at least one of the following: neck pain, nuchal rigidity, altered cutaneous sensations, photophobia, or visual disturbances. RESULTS: Twelve travelers met the case definition for eosinophilic meningitis. The symptoms began a median of 11 days (range, 6 to 31) after their return to the United States. Eosinophilia was eventually documented in all nine patients who were hospitalized, although on initial evaluation, it was present in the peripheral blood of only four of the nine (44 percent) and in the cerebrospinal fluid of five (56 percent). Repeated lumbar punctures and corticosteroid therapy led to improvement in symptoms in two of three patients with severe headache, and intracranial pressure decreased during corticosteroid therapy in all three. Consumption of one meal (P=0.001) and of a Caesar salad at that meal (P=0.007) were strongly associated with eosinophilic meningitis. Antibodies against an A. cantonensis--specific 31-kD antigen were detected in convalescent-phase serum samples from 11 patients. CONCLUSIONS: Among travelers at risk, the presence of headache, elevated intracranial pressure, and pleocytosis, with or without eosinophilia, particularly in association with paresthesias or hyperesthesias, should alert clinicians to the possibility of A. cantonensis infection. PMID- 11870245 TI - Images in clinical medicine. Syphilitic aortitis. PMID- 11870246 TI - Clinical practice. Aortic stenosis. PMID- 11870247 TI - Imatinib mesylate--a new oral targeted therapy. PMID- 11870248 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 7-2002. A 47-year-old woman with late recurrent hyperparathyroidism. PMID- 11870249 TI - Proteolytic antibodies against factor VIII in hemophilia A. PMID- 11870250 TI - Angiotensin-receptor blockers, type 2 diabetes, and renoprotection. PMID- 11870251 TI - Beta-blockade and severe burns. PMID- 11870252 TI - Shipped and locally transplanted renal allografts. PMID- 11870253 TI - Essential tremor. PMID- 11870254 TI - AIDS--past and future. PMID- 11870255 TI - Tourette's syndrome. PMID- 11870256 TI - Combined aortic surgery and implantation of a left ventricular assist device. PMID- 11870257 TI - Adverse events after imatinib mesylate therapy. PMID- 11870258 TI - Protecting research subjects--the crisis at Johns Hopkins. PMID- 11870259 TI - Neurobiology of relapse to heroin and cocaine seeking: a review. AB - The objective of this article is to review data from studies that used a reinstatement model in rats to elucidate the neural mechanisms underlying relapse to heroin and cocaine seeking induced by exposure to the self-administered drug (drug priming), conditioned drug cues, and stressors. These factors were reported to contribute to relapse to drug use in humans following prolonged abstinence periods. In the reinstatement model, the ability of acute exposure to drug or nondrug stimuli to reinstate drug seeking is determined following training for drug self-administration and subsequent extinction of the drug-reinforced behavior. We will review studies in which pharmacological agents were injected systemically or intracranially to block (or mimic) reinstatement by drug priming, drug cues, and stressors. We also will review studies in which brain lesions, in vivo microdialysis and electrochemistry, and gene expression methods were used to map brain sites involved in relapse to drug seeking. Subsequently, we will discuss theoretical issues related to the processes underlying relapse to drugs and address methodological issues in studies on reinstatement of drug seeking. Finally, the implications of the findings from the studies reviewed for addiction theories and treatment will be discussed. The main conclusion of this review is that the neuronal mechanisms involved in relapse to heroin and cocaine seeking induced by drug priming, drug cues, and stressors are to a large degree dissociable. The data reviewed also suggest that the neuronal events mediating drug-induced reinstatement are to some degree dissociable from those mediating drug reinforcement. PMID- 11870260 TI - Fine tuning of sympathetic transmitter release via ionotropic and metabotropic presynaptic receptors. AB - The release of transmitters at sympathoeffector junctions is not constant, but subject to modulation by a plethora of different mechanisms. In this respect, presynaptic receptors located on the sympathetic axon terminals are of utmost importance, because they are activated by exogenous agonists and by endogenous neurotransmitters. In the latter case, the transmitters that activate the presynaptic receptors of a nerve terminal may be released either from the very same nerve ending or from a different axon terminal, and the receptors involved are auto- and heteroreceptors, respectively. In terms of their structural and functional features, receptors of sympathetic axon terminals can be categorized as either ionotropic (transmitter-gated ion channels) or metabotropic (most commonly G protein-coupled) receptors. This review summarizes results on more than 30 different metabotropic and four different ionotropic receptors that have been found to control the amount of transmitter being released from sympathetic neurons. Each of these receptors may not only stimulate, facilitate, and reduce sympathetic transmitter release, respectively, but also interact with the functions of other receptors present on the same axonal varicosity. This provides a multitude of mechanisms that regulate the amount of sympathetic transmitter output. Accordingly, a sophisticated cross-talk within and between extra- and intracellular signals is integrated at axon terminals to adapt the strength of sympathoeffector transmission to a given situation. This will not only determine the function of the sympathetic nervous system in health and disease, but also therapeutic and untoward effects of drugs that bind to the presynaptic receptors in sympathetically innervated tissues. PMID- 11870261 TI - Mitochondria as a pharmacological target. AB - Mitochondria play a central role in energy metabolism within the cell. Mitochondrial dysfunctions lead to various neurodegenerative disorders and to the so-called "mitochondrial diseases". A vast amount of evidence points to the implication of mitochondria in such complex processes as apoptosis and cardioprotection. The purpose of this review is to present a recent state of our knowledge and understanding of the action of various therapeutically applied substances on mitochondria. These include antitumor, immunosuppressant, and antiviral drugs, potassium channel openers, sulfonylureas, and anesthetics. Some of these substances are specifically designed to affect mitochondrial functions. In other cases, drugs with primary targets in other cellular locations may modify mitochondrial functions as side effects. In any case, identification of mitochondria as primary or secondary targets of a drug may help us to better understand the drug's mechanism of action and open new perspectives for its application. As far as possible, the molecular mechanisms of the interference of particular drugs in the mitochondrial metabolism will be described. In some cases, metabolic routes in which the drugs interfere will also be briefly outlined. PMID- 11870262 TI - Liver-enriched transcription factors in liver function and development. Part I: the hepatocyte nuclear factor network and liver-specific gene expression. AB - Numerous studies have established the pivotal role of liver-enriched transcription factors in organ development and cellular function, and there is conclusive evidence for transcription factors to act in concert in liver-specific gene expression. During organ development and in progenitor cells the timely expression of certain transcription factors is necessary for cellular differentiation, and there is overwhelming evidence for hierarchical and cooperative principles in a networked environment of transcription factors. The search for molecular switches that control stem cell imprinting and liver specific functions has lead to the discovery of many interactions between such different molecules as transcription factors, coactivators, corepressors, enzymes, DNA, and RNA. Many of these interactions either repress or activate liver-specific gene expression. It thus can be demonstrated that specific mutational changes in liver-enriched transcription factors lead to altered intermolecular interactions with the consequence of human disease. This review provides an overview of our current knowledge about liver-enriched transcription factors and their role in liver function and development. We review the basic principles of gene transcription, the role of liver-enriched transcription factors in liver gene regulation, and the classification of transcription factors by their DNA-binding domains. PMID- 11870264 TI - Combination antiplatelet therapy following brachytherapy with restenting: it ain't over 'til the fat lady sings. PMID- 11870263 TI - Late stent thrombosis in brachytherapy: the role of long-term antiplatelet therapy. AB - Advances in percutaneous coronary intervention (PCI) have emerged in the past decade. Stenting has improved upon the limitations of angioplasty, acute vessel closure and restenosis by providing mechanical vascular support, resulting in sustained clinical and angiographic benefit. This has led to greater utilization of the technique, although it is associated with a significant incidence of in stent restenosis. Neointimal hyperplasia is the pathophysiologic process that leads to in-stent restenosis. Brachytherapy can be effective in reducing the occurrence of this process. Unfortunately, brachytherapy trials have identified the phenomenon of late stent thrombosis as a potentially serious complication of this procedure. Late stent thrombosis is thrombosis that occurs > 30 days after PCI. The risk of thrombosis is increased in patients receiving a new stent in addition to brachytherapy. It also appears to be increased when adjunctive antiplatelet therapy with ticlopidine or clopidogrel is discontinued early. Strategies to prevent late stent thrombosis include the prolonged use of combination antiplatelet therapy in addition to limited placement of new stents in patients treated with brachytherapy for in-stent restenosis. PMID- 11870265 TI - Relationship between pressure-derived collateral blood flow and diabetes mellitus in patients with stable angina pectoris: a study based on coronary pressure measurement. AB - Although the pressure gradient between the normal and stenotic vascular regions is known to be the most important factor for collateral vessel development, factors which are responsible for variations among patients with ischemic heart disease are not well known. Likewise, it is still not clear whether diabetes mellitus (DM) has any effect on coronary collateral development. Coronary angiography, the most commonly used technique for studying collateral circulation, may not be accurate in assessing collateral circulation because most collaterals are situated intramurally or are too small to visualize angiographically. Intracoronary pressure measurement is a new technique to provide accurate and quantitative information about the collateral circulation. Therefore, we sought the effects of DM on coronary collateral vessels in patients with coronary artery disease by using intracoronary pressure measurement technique. METHODS: Study material consisted of 40 patients (20 diabetic) with chronic ischemic heart disease referred to angiography laboratory due to their ischemic symptoms verified previously with at least one non-invasive test. All of the patients had single vessel disease with more than 70% narrowing and had undergone PTCA and/or stent implantation procedure for this vessel. Quantitative coronary angiographic analysis (QCA) and all of the coronary pressure measurements were performed both pre- and post-revascularization procedure. After angiography, a fiber-optic pressure monitoring guidewire (Pressure wire, RADI Medical Systems, Inc., Reading, Massachusetts) was advanced to the stenosis to be dilated. The same wire was used as a guidewire for the angioplasty catheter. During complete occlusion with balloon inflation, distal pressure was recorded as coronary wedge pressure (CWP). As a more valuable parameter, collateral flow index (CFI) was determined by the ratio of simultaneously measured CWP (mmHg) to mean aortic pressure (Pa, mmHg, obtained from the guiding catheter) (CFI: CWP/Pa). RESULTS: Pressure measurements were performed on 20 diabetic and 20 non diabetic patients. The mean value of CWP was 18.1 8.6 mmHg in the diabetic group and 26.8 +/- 9.6 mmHg in the non-diabetic group; this difference was statistically significant (p < 0.01). Also, the mean value of CFI was significantly higher in the non-diabetic group (0.17 +/- 0.08 in the diabetic group and 0.25 0.09 in the non-diabetic group; p < 0.01). CONCLUSION: This study demonstrated that the coronary collateral vessel development is impaired in diabetic patients compared with non-diabetic patients. PMID- 11870266 TI - Coronary collaterals and their assessment. PMID- 11870267 TI - The relationship between corrected TIMI frame count and myocardial fractional flow reserve. AB - Thrombolysis in myocardial infarction (TIMI) frame count (TFC) is a reproducible, objective and quantitative index of coronary flow that allows standardization of TIMI flow grades. After myocardial infarction (MI), hyperemic and basal flows decrease in the infarct region due to residual stenosis and increased resistance in the microvasculature subtended by this infarct-related artery (IRA). Myocardial fractional flow reserve (FFRmyo) is a lesion-specific index for epicardial stenosis. FFRmyo can also be used as a criterion for assessing the success of revascularization therapy. The aim of this study was to investigate the relationship between improvement of the corrected TFC (CTFC) and change in FFRmyo, which were determined in the IRA in patients with recent MI who underwent mechanical revascularization. METHODS: Forty-one patients (28 male; age, 58.3 +/- 12.3 years) early after MI with > 60% narrowing in the IRA who underwent stent implantation for this residual stenosis were included in this study. After angiography, a fiber-optic pressure monitoring guidewire was positioned distal to the stenosis to be dilated. Means of aortic (Pa) and distal (Pd) pressures were recorded simultaneously under baseline and hyperemic conditions. FFRmyo (Pd/Pa) was determined under adenosine hyperemia before and after stent implantation, as was TFC. Percentage of the improvement in the CTFC and FFRmyo was calculated for each lesion. The first frame used for TFC was defined by a column of contrast extending across > 70% of the arterial lumen and last frame counted was that in which contrast first appeared in the distal, pre-defined landmark branches for each vessel. CTFC was calculated for the LAD by dividing TFC of the LAD by a factor of 1.7. RESULTS: Eighteen patients had LAD, 10 had CX and 13 had RCA lesions. After stent implantation, the CTFC improved by 41% (from 32.3 +/- 3.4 to 19.2 +/- 2.1; p < 0.0001) and FFRmyo improved by 69% (from 0.53 +/- 0.11 to 0.90 +/- 0.15; p < 0.0001). There was a statistically significant correlation between the change in FFRmyo and improvement in the TFC (r = 0.85). CONCLUSION: Improvement in CTFC is associated with change in FFRmyo in the IRA undergoing mechanical revascularization. After mechanical revascularization, CTFC decreases proportionally to improvement of the FFRmyo. These two parameters support each other in terms of successful revascularization and this relationship shows the recruitability of CTFC in terms of FFRmyo as well. PMID- 11870268 TI - New techniques for the evaluation of the vulnerable plaque. PMID- 11870269 TI - Sidebranch compromise during percutaneous coronary interventions. AB - Justified concerns exist about coronary balloon angioplasty and stent deployment when a sidebranch is within the vicinity of the interventional site. Assessment of the jeopardized sidebranch for the risk of compromise can be made by evaluating the sidebranch diameter, the myocardial territory supplied, the relationship to the parent vessel lesion and the presence of ostial disease. This can help in the decision-making process of the proposed intervention, allowing a strategy to be pre-planned in the event of sidebranch compromise. Sidebranch compromise associated with flow reduction in a branch vessel that is of medium or large diameter and serving moderate or large territory is often associated with a cardiac enzyme rise. It is now recognized that any cardiac enzyme rise after intervention is associated with increased long-term risk and such compromised sidebranches may be considered for re-opening to help preserve the myocardium. A variety of balloon angioplasty and stenting techniques are described for sidebranch compromise with some short-term success. Long-term outcomes and effect of sidebranch intervention have not yet been fully evaluated. PMID- 11870270 TI - Transradial renal artery angioplasty and stenting. AB - Transradial arterial access is an alternative approach for coronary interventions. The utilization of these cardiac systems may facilitate endovascular treatment of other vascular territories. This report describes our first experience with percutaneous transluminal renal artery angioplasty and stenting (PTRAS) using the transradial approach. This case demonstrates the feasibility of the radial approach to treat severe renal artery disease safely with PTRAS. Comparison with femoral and brachial arterial access sites is beyond the scope of this paper, but the cardiac literature has demonstrated the safety and efficacy of transradial artery access. With refinement of the endovascular equipment, the radial approach could become an attractive alternative entry site for renal artery interventions. PMID- 11870271 TI - Transcatheter endovascular therapy of a traumatic common hepatic artery aneurysm. AB - Dilemma was encountered regarding diagnosis and treatment when a farmer presented with a progressively increasing pulsatile abdominal mass and signs of obstructive jaundice. A computerized tomographic scan reported it to be a pancreatic pathology, but angiographically it was found to be a large aneurysm arising from the common hepatic artery. Surgery was thought to be highly risky; therefore, a transcatheter technique was successfully applied to tackle this life-threatening situation. PMID- 11870272 TI - Interview with Frank J. Criado [interview by Laurie Gustafson]. PMID- 11870273 TI - High-temperature synthesis of Rb2MnP2S6 in molten salt medium. AB - Transparent yellow plates of rubidium manganese hexathiodiphosphate, Rb(2)MnP(2)S(6), were synthesized in molten RbBr. The compound is isotypic to other compounds of the type A(2)MP(2)Q(6) (A = K, Rb, Cs; M = Mn, Fe; Q = S, Se). Its structure can be viewed as columns of face-sharing S(6) polyhedra parallel to the a axis, interconnected by Rb(+). The S(6) polyhedra are centered alternately by Mn (in octahedral coordination) and P(2) units (in trigonal antiprisms). The Mn atom and P(2)S(6) group lie on centers of symmetry. PMID- 11870274 TI - Barium tetraphosphate. AB - The structure of the low-temperature form of barium tetraphosphate, Ba(3)P(4)O(13), shows the tetraphosphate to exist in an S conformation. PMID- 11870275 TI - A tetragonal polymorph of caesium hydroxide monohydrate, CsOH x H2O, from X-ray powder data. AB - Tetragonal caesium hydroxide monohydrate, CsOH x H(2)O, a clathrate hydrate, is a polymorph of three known hexagonal or pseudo-hexagonal modifications. It was obtained as a by-product in a high-pressure experiment. Whether it is a high pressure polymorph, however, remains to be verified. The Cs atoms are situated in cavities of the form of a bicapped pentagonal prism, within an infinite three dimensional hydrogen-bonded oxygen framework that is locally identical to layers found in the hexagonal modifications. The Cs atom and one of the two H atoms are at sites with (macro)4m2 symmetry, the O atom has mm site symmetry and the second H atom has 2/m symmetry. PMID- 11870276 TI - Strontium manganese diselenite, SrMn(SeO(3))(2), containing unusual MnO(5+1) polyhedra. AB - Hydrothermally prepared SrMn(SeO(3))(2) contains infinite chains of vertex sharing irregular MnO(5+1) polyhedra [mean Mn-O 2.226(3)A], which are fused into layers via pyramidal SeO(3) groups [mean Se-O 1.698(3)A]. Nine-coordinate Sr(2+) cations [mean Sr-O 2.715(4)A] complete the layered structure. PMID- 11870277 TI - HgWO(4) synthesized at high pressure and temperature. AB - Crystals of mercury(II) tungstate(VI), HgWO(4), were grown in sealed gold tubes under an Ar atmosphere at 300 MPa and 973 K. The monoclinic crystal structure (C2/c) of HgWO(4) consists of zigzag chains of edge-sharing WO(6) octahedra running along the c axis and layers of very distorted corner-sharing HgO(6) octahedra in the bc plane. The Hg atom lies on an inversion centre and the W atom is on a twofold axis. No structural effects which can be ascribed to the high pressure used in the synthesis were found. PMID- 11870278 TI - Hg(3)AlF(6)O(2)H, a structure with a short interpolyhedral O...O distance. AB - The crystal structure of Hg(3)AlF(6)O(2)H, trimercury(II) aluminium hydrogen hexafluoride dioxide, can be derived from a slightly distorted cubic close-packed (ccp) arrangement of the metal atoms, where three quarters of the positions are occupied by Hg atoms and one quarter by Al atoms. The F and O atoms are considerably dislocated from the tetrahedral voids of this arrangement, thus forming [HgO(2)F(6)] polyhedra, with two short Hg-O distances, two intermediate Hg-F distances and four longer Hg-F distances, and nearly ideal [AlF(6)] octahedra. The H atoms are presumably located close to the inversion centre. Their positions were derived from crystal chemical arguments, and they take part in the formation of O-H.O hydrogen bonds between two O atoms, with an O.O distance of 2.562 (9) A. PMID- 11870279 TI - Synchrotron X-ray study of orthorhombic Rb(3)Ta(5)O(14) with a modified pyrochlore structure. AB - The structure of a new orthorhombic trirubidium pentatantalum oxide (Rb(3)Ta(5)O(14)) phase with Pnma symmetry was identified from a half sphere of synchrotron X-ray data measured at a wavelength of 0.85 A. This notionally linked TaO(6) octahedral structure broadly consists of three different modifications of the pyrochlore ring motif with layer stacking normal to (205) planes. Successive pyrochlore layers do not simply stack normal to these planes but are offset along the [100] axis. An unusual aspect of this structure is the occurrence of TaO(5) trigonal bipyramids in structurally complex regions where the modified pyrochlore rings connect. PMID- 11870280 TI - Chloroheptakis(dimethyl sulfoxide)uranium(IV) trichloride. AB - In the title complex, [UCl(C(2)H(6)OS)(7)]Cl(3), the uranium metal center is coordinated in a distorted bicapped trigonal prism geometry by seven O atoms from dimethyl sulfoxide ligands and by a terminal chloride ligand. Charge balance is maintained by three outer-sphere chloride ions per uranium(IV) metal center. Principle bond lengths include U-O 2.391 (2)-2.315 (2) A, U-Cl 2.7207 (9) A, and average S-O 1.540 (5) A. PMID- 11870281 TI - Pseudosymmetry in pyridinium tetrachloro(oxo)pyridineniobate(V) pyridine solvate. AB - The title compound, (C(5)H(6)N)[NbCl(4)O(C(5)H(5)N)]center dotC(5)H(5)N, crystallizes as discrete ions, with a very strong linear N-H...N hydrogen-bonding interaction between the cation and the solvate pyridine molecule [N...N 2.755 (5) A]. All chemical species occupy crystallographic twofold axes. The ligated and solvate pyridines form ABABAB stacks in the lattice. There is pseudosymmetry which emulates a centred unit cell in Amm2, but it is not supported by the diffraction pattern, which is consistent with the correct space group Pnc2. Three crystallographic software packages suggested space group Amm2 over Pnc2, while a fourth indicated Pnc2, a subgroup of Amm2. PMID- 11870282 TI - A liquid crystal derived from ruthenium(II,III) and a long-chain carboxylate. AB - The title compound, catena-poly[[tetrakis(mu-decanoato kappa(2)O:O')diruthenium(II,III)(Ru-Ru)]-mu-octanesulfonato-kappa(2)O:O'], [Ru(2)(C(10)H(19)O(2))(4)(C(8)H(17)O(3)S)], is an octanesulfonate derivative of the mixed-valence complex diruthenium tetradecanoate. The equatorial carboxylate ligands are bidentate, bridging two Ru atoms to form a dinuclear structure. Each of the two independent dinuclear metal complexes in the asymmetric unit is located at an inversion centre. The octanesulfonate anion bridges the two dinuclear units through axial coordination. The alkyl chains of the carboxylate and sulfonate ligands are arranged in a parallel manner. The global structure can be seen as infinite chains of polar moieties separated by a double layer of non polar alkyl groups, without interdigitation of the alkyl chains. PMID- 11870283 TI - Bis(8-quinolinolato-N,O)platinum(II) and its synthetic intermediate, 8 hydroxyquinolinium dichloro(8-quinolinolato-N,O)platinate(II) tetrahydrate. AB - Bis(8-quinolinolato-N,O)platinum(II), [Pt(C(9)H(6)NO)(2)], (I), has a centrosymmetric planar structure with trans coordination. The molecules form an inclined pi stack, with an interplanar spacing of 3.400 (6) A. 8 Hydroxyquinolinium dichloro(8-quinolinolato-N,O)platinate(II) tetrahydrate, (C(9)H(8)NO)[PtCl(2)(C(9)H(6)NO)] x 4H(2)O, (II), is soluble in water and is regarded as the synthetic intermediate of the insoluble neutral compound (I). The uncoordinated 8-hydroxyquinolinium cations and the monoquinolinolate complexes form an alternating pi stack. The origins of fluorescence and phosphorescence in (II) are assigned to the 8-hydroxyquinolinium cation and the monoquinolinolate- Pt complex, respectively. PMID- 11870284 TI - The sodium salt of 2-hydroxy-5-nitrobenzylsulfonic acid. AB - The structural data for sodium 2-hydroxy-5-nitrobenzylsulfonate monohydrate, Na(+) x C(7)H(6)NO(6)S(-) x H(2)O, which mimics an artificial substrate for human arylsulfatase A, viz. p-nitrocatechol sulfate, reveal that the geometric parameters of the substrate and its analogue are very similar. Two water molecules, the phenolic O atom and three sulfonate O atoms form the coordination sphere of the Na(+) ion, which is a distorted octahedron. The Na(+) cations and the O atoms join to form a chain polymer. PMID- 11870285 TI - Interpenetrating nets in cis-bis(pyridine-4-carboxylate)nickel(II). AB - The title compound, [Ni(C(6)H(4)NO(2))(2)], crystallized in a three-dimensional framework consisting of three interpenetrating diamond-like nets. The Ni atom is on a twofold axis, the coordination is '(O(2))(2)N(2)' in a cis arrangement and the ligands are bridging. PMID- 11870286 TI - Aquachloro(1,10-phenanthroline-kappa 2N,N')(p-toluenesulfonato-kappa O)copper(II). AB - In the title compound, [CuCl(C(7)H(7)O(3)S)(C(12)H(8)N(2))(H(2)O)], the central Cu atom is coordinated by a water molecule, a chloride ion, an O-monodentate p toluenesulfonate anion and an N,N'-bidentate 1,10-phenanthroline ligand. The copper environment is best described as a slightly distorted square pyramid, with bond distances Cu-Cl 2.2282 (9) A, Cu-OW 1.984 (3) A, and Cu-N 2.006 (3) and 2.028 (3) A; the apical Cu-O distance is 2.281 (2) A. In the supramolecular structure, pi-pi-stacking stabilization is observed, and classical and non classical hydrogen bonds also play an important role. PMID- 11870288 TI - Chlorobis(triphenylphosphine)nickel(I). AB - Crystals of the title compound, [NiCl(C(18)H(15)P)(2)], contain one molecule per asymmetric unit with no short intermolecular interactions. This is noteworthy since previous studies have reported that the formally 15-electron species oligomerizes in the solid state. The nickel(I) centre has a distorted trigonal planar coordination geometry, the origin of which is suggested to be electronic in nature. PMID- 11870287 TI - Trichloro(1,1-dimethylbiguanidium-kappa N3)zinc(II). AB - The title compound, [ZnCl(3)(C(4)H(12)N(5))], was prepared from aqueous solution and its structure determined. The coordination geometry around the Zn atom is a tetrahedron, with the central Zn atom bound to three Cl atoms and to one N atom of the biguanide ligand. The dihedral angle between the two guanidine groups is 67.86 (1). PMID- 11870289 TI - Bis[1,1'-bis(diphenylphosphino)ferrocenium] hexadecachlorotetraantimony(III) ethanol solvate. AB - In the title compound, [Fe(C(17)H(14)P)(2)](2)[Sb(4)Cl(16)] x C(2)H(6)O, the Fe atoms lie on inversion centres and the pairs of cyclopentadienyl rings are consequently in a fully staggered conformation. The centrosymmetric anionic clusters formed by [Sb(4)Cl(16)](4-) are surrounded by the cations and are held together by weak C-H...Cl interactions. These formations stack along the a axis to form columns, and the columns are interconnected by another weak C-H...Cl interaction along the b axis. PMID- 11870290 TI - A pseudo-cage complex of silver(I) with tripodal tetraamine having benzyl end groups, [tris[2-(benzylamino)ethyl]amine-kappa 4N]silver(I) perchlorate. AB - The synthesis and crystal structure of [tris[2-(benzylamino)ethyl]amine kappa(4)N]silver(I) perchlorate, [Ag(C(27)H(36)N(4))]ClO(4) or [Ag(bz(3)tren)]ClO(4) [bz(3)tren is tris[2-(benzylamino)ethyl]amine or N,N',N'' tribenzyltris(2-aminoethyl)amine] are reported. The Ag atom is coordinated to four N atoms of the tren unit and is located 0.604 (3) A out of the trigonal plane described by the three secondary amine N atoms, away from the bridgehead N atom. Edge-to-face pi-pi interactions between the aromatic end groups, and weak interactions between Ag and arene, allow the formation of a pseudo-cage complex. PMID- 11870291 TI - Three different bonding modes of cyano groups in the coordination polymer [Cu(en)2(H(2)O)][Cu(en)(2)Ni(2)Cu(2)(CN)(10)] x 2H(2)O (en is 1,2-diaminoethane). AB - Partial reduction of the Cu(II) ions in the aqueous system Cu(II)-en [Ni(CN)(4)](2-) (1/1/1) (en is 1,2-diaminoethane) yields a novel heterobimetallic mixed-valence compound, poly[[aquabis(1,2-diaminoethane)copper(II)] [hexa-mu cyano-tetracyanobis(1,2-diaminoethane)tricopper(I,II)dinickel(II)] dihydrate], [Cu(C(2)H(8)N(2))(2)(H(2)O)][Ni(2)Cu(3)(CN)(10)(C(2)H(8)N(2))(2)] x 2H(2)O or [Cu(en)(2)(H(2)O)][Cu(en)(2)Ni(2)Cu(2)(CN)(10)] x 2H(2)O. The structure is formed by a negatively charged two-dimensional array of the cyano complex [Cu(en)(2)Ni(2)Cu(2)(CN)(10)](n)(2n-), [Cu(en)(2)(H(2)O)](2+) complex cations and water molecules of crystallization. These last are involved in a complicated hydrogen-bonding system. The cyano groups act as terminal, mu(2)-bridging or mu(3)-bridging ligands. PMID- 11870292 TI - Decacarbonylbis(methylcyclopentadienyl)-tetrahedro-diiridiumdimolybdenum and decacarbonylbis(tetramethylcyclopentadienyl)-tetrahedro-diiridiumdimolybdenum dichloromethane hemisolvate. AB - The two title compounds, [Mo(2)Ir(2)(C(6)H(7))(2)(CO)(10)] and [Mo(2)Ir(2)(C(9)H(13))(2)(CO)(10)] x 0.5CH(2)Cl(2), respectively, or collectively [Mo(2)Ir(2)(mu-CO)(3)(CO)(7)(eta(5)-C(5)H(5-n)Me(n))(2)] (n = 1 or 4), have a pseudo-tetrahedral Mo(2)Ir(2) core geometry, an eta(5)-C(5)H(5-n)Me(n) group ligating each Mo atom, bridging carbonyls spanning the edges of an MoIr(2) face and seven terminally bound carbonyl groups. PMID- 11870293 TI - catena-Poly[[di-mu-chloro-bis[(triphenylphosphine)silver(I)]]-mu-2 aminopyrimidine-kappa 2N1:N3]. AB - In the title supramolecular complex, [Ag(2)Cl(2)(C(4)H(5)N(3))(C(18)H(15)P)(2)](n), a one-dimensional chain is formed by dimeric [Ag(2)Cl(2)(PPh(3))(2)] units bridged by 2-aminopyrimidine moieties. The Ag atoms are four-coordinate, with an AgCl(2)NP core. A crystallographic inversion centre is located in the centre of the Ag(2)Cl(2) chelate ring, while the crystallographic twofold axis bisects the 2-aminopyrimidine ligand. PMID- 11870294 TI - The trigonal-bipyramidal triiodothallium(III) complex [TlI(3)[(CH(3))(2)SO]2]. AB - The title compound, bis(dimethyl sulfoxide)triiodothallium(III), [TlI(3)(C(2)H(6)OS)(2)], was crystallized from equimolar amounts of Tl(I)I and I(2) in a dimethyl sulfoxide (DMSO) solution. After the initial redox reaction, the thallium(III)-iodo complex forms and precipitates as a DMSO solvate. In the crystal structure, Tl is surrounded by three iodide ligands in the equatorial plane and two O-coordinated DMSO molecules in the axial positions, forming a slightly distorted trigonal bipyramid. The complex lies on a twofold rotation axis, making the DMSO molecules and two of the I atoms crystallographically equivalent. PMID- 11870295 TI - Aqua[bis(pyrimidin-2-yl-kappa N)amine](carbonato-kappa 2O,O')copper(II) dihydrate. AB - The title mononuclear complex, [Cu(CO(3))(C(8)H(7)N(5))(H(2)O)] x 2H(2)O, was obtained by fixation of CO(2) by a mixture of copper(II) tetrafluoroborate and the ligand bis(pyrimidin-2-yl)amine in ethanol/water. The Cu(II) ion of the complex has a distorted square-pyramidal environment, with a basal plane formed by two N atoms of the ligand and two chelating O atoms of the carbonate group, while the apical position is occupied by the O atom of the coordinating water molecule. In the solid state, hydrogen-bonding interactions are dominant, the most unusual being the Watson-Crick-type coplanar ligand pairing through two N- H...N bonds. Lattice water molecules also participate in hydrogen bonding. PMID- 11870296 TI - A copper(II)-pyrazole complex cation with (macro)3 imposed symmetry. AB - The reaction of Cu(ClO(4))(2) x 6H(2)O, NaAsF(6) and excess pyrazole yields hexakis(pyrazole-kappa N(2))copper(II) bis(hexafluoroarsenate), [Cu(C(3)H(4)N(2))(6)](AsF(6))(2) or [Cu(pzH)(6)](AsF(6))(2) (pzH is pyrazole), (I). The analogous hexakis(pyrazole-kappa N(2))copper(II) hexafluorophosphate perchlorate complex, [Cu(C(3)H(4)N(2))(6)](PF(6))(1.29)(ClO(4))(0.71) or [Cu(pzH)(6)](PF(6))(1.29)(ClO(4))(0.71), (II), is obtained in a similar fashion, using KPF(6) in place of NaAsF(6). Both compounds contain the hitherto unknown [Cu(pzH)(6)](2+) complex cation, in which the copper(II) ion lies at the center of a regular octahedron of coordinated N atoms. The cation has crystallographically imposed -3 symmetry. The X-ray data indicate that the lack of the expected distortion can be accounted for by the presence of either static Jahn-Teller disorder or dynamic Jahn-Teller distortion. PMID- 11870297 TI - Tris(1-ethyl-3-methylimidazolium) hexachlorolanthanate. AB - In the title complex, (C(6)H(11)N(2))(3)[LaCl(6)], centrosymmetric octahedral hexachlorolanthanate anions are located at the corners and face-centers of the monoclinic unit cell. The ring H atoms of the cations interact with the Cl atoms of the anions via hydrogen bonding, and bifurcation of the hydrogen bonding is observed. Cation-cation interactions via hydrogen bonding between the ring H atoms and pi-electrons of aromatic rings are also observed as in other imidazolium salts. PMID- 11870299 TI - catena-Poly[[bis(nicotinamide-kappa N1)copper(I)]-mu-thiocyanato-kappa 2N:S]. AB - The Cu(I) cations in the title compound, [Cu(NCS)(C(6)H(6)N(2)O)(2)](n), are coordinated by N atoms from each of two mirror-related nicotinamide ligands, as well as by one N atom of one thiocyanate ligand and one S atom of a symmetry related thiocyanate ligand, within a slightly distorted tetrahedron. The Cu(I) cations and the thiocyanate anions are located on a crystallographic mirror plane and the nicotinamide ligands occupy general positions. The Cu(I) cations are connected by the thiocyanate anions to form chains in the direction of the crystallographic a axis. These chains are connected by hydrogen bonds between the amide H atoms and the O atoms of adjacent nicotinamide ligands, to give a three dimensional structure. PMID- 11870298 TI - Tetrakis[(4-aminopyridinio)acetato-kappa O]diaquamanganese(II) diperchlorate. AB - In the centrosymmetric title complex, [Mn(C(7)H(8)N(2)O(2))(4)(H(2)O)(2)](ClO(4))(2), the Mn(II) ion is in an octahedral environment, with the equatorial plane being defined by the O atoms of four monodentate carboxylate groups, and the octahedron being completed by two trans-coordinated water molecules. There are intramolecular hydrogen bonds between the coordinated water molecules and the non-coordinated O atoms of the carboxylate groups. Hydrogen bonds between the amino groups and the carboxylate groups of neighbouring molecules generate a layered hydrogen-bonded network. PMID- 11870300 TI - Three heterotrinuclear Schiff base complexes of nickel(II) with cobalt(II), copper(II) and manganese(II). AB - The title compounds, bis(dimethylformamide)-1 kappa O,3 kappa O-bis[mu-2,2'-[2,2' dimethylpropane-1,3-diylbis(nitrilomethylidyne)]diphenolato]-1 kappa(4)N,N',O,O':2 kappa(2)O,O';2 kappa(2)O,:3 kappa(4)N,N'prime prime or minute,O,O'-di-mu-nitrito-1:2 kappa(2)N:O;2:3 kappa(2)O:N-dinickel(II)cobalt(II), [CoNi(2)(NO(2))(2)(C(19)H(22)N(2)O(2))(2)(C(3)H(7)NO)(2)], (I), -copper(II), [CuNi(2)(NO(2))(2)(C(19)H(22)N(2)O(2))(2)(C(3)H(7)NO)(2)], (II), and manganese(II), [MnNi(2)(NO(2))(2)(C(19)H(22)N(2)O(2))(2)(C(3)H(7)NO)(2)], (III), consist of centrosymmetric linear heterotrinuclear metal complexes. The three complexes are isostructural. There are three bridges across the Ni-M atom pairs (M is Co(2+), Cu(2+) or Mn(2+)) in each complex, involving two O atoms of a mu N,N'-bis(salicylidene)-2,2'-1,3-propanediaminate ligand and an N-O moiety of a mu nitrito group. The coordination sphere around each metal atom, whether Co(2+), Cu(2+), Mn(2+) or Ni(2+), can be described as distorted octahedral. The Ni...M distances are 2.9988 (5) A in (I), 2.9872 (5) A in (II) and 3.0624 (8) A in (III). PMID- 11870301 TI - catena-Poly[bis[tetraaquacobalt(II)-mu-4,4'-bipyridine-kappa 2N:N'] benzene 1,2,4,5-tetracarboxylate(4-) dihydrate]. AB - The title compound, [Co(C(10)H(8)N(2))(H(2)O)(4)](2)(C(10)H(2)O(8)) x 2H(2)O, consists of two crystallographically independent Co(II) atoms linked by 4,4' bipyridine ligands into one-dimensional chains, which are further connected into a three-dimensional framework linked by [C(6)H(2)(COO)(4)](4-) anions and water molecules, achieved through complex hydrogen bonding. PMID- 11870302 TI - The binary adduct of 3,6,9,16,19,22-hexaazatricyclo[22.2.2.211,14]triaconta 11,13,24,26(1),27,29-hexaene and benzene-1,2,4,5-tetracarboxylic acid. AB - The crystals of the title salt, 6,21-diaza-3,9,18,24 tetraazoniatricyclo[22.2.2.2(11,14)]triaconta-11,13,24,26(1),27,29-hexaene benzene-1,2,4,5-tetracarboxylate(4-) hexahydrate, C(24)H(42)N(6)(4+) x C(10)H(2)O(8)(4-) x 6H(2)O, are formed by the intermolecular interaction of a macrocyclic hexamine with a molecule of C(6)H(2)(COOH)(4) in aqueous solution. Both the cation and the anion are on inversion centres. Hydrogen bonds are formed between the four ammonium cations in the hexamine and the four carboxylate anions in the aromatic acid. Stacks exist along the crystallographic a axis in the solid state. The water molecules also take part in a hydrogen-bonding network which joins these stacks together. PMID- 11870303 TI - Stigmasterol hemihydrate. AB - The title compound, stigmasta-5,22-dien-3beta-ol hemihydrate, C(29)H(48)O x 0.5H(2)O, previously thought to be the monohydrate, has two sterol molecules and one water molecule in the asymmetric unit. In both sterol molecules, the methyl group of the ethyl substituent at the end of the hydrocarbon chain is disordered over two sites. The OH group of molecule A donates a hydrogen bond to a water molecule and accepts a hydrogen bond from the OH group of molecule B. The OH group of molecule B accepts two hydrogen bonds from water molecules. PMID- 11870304 TI - 3,4-Dihydro-6-methyl-3-beta-D-ribofuranosyl-4,5'-cyclo-9H-imidazo[1,2-a]purin-9 one hydrate. AB - In the title compound, C(13)H(13)N(5)O(4) x H(2)O (4,5'-cyclowyosine x H(2)O), the cyclization forces a syn arrangement of the aglycon with respect to the sugar moiety. The ribofuranose part of the molecule displays a beta-D configuration with an envelope C1'-endo pucker. The molecules are arranged in columns along the short a axis and are linked to water molecules through O-H...O and O-H...N hydrogen bonds. PMID- 11870305 TI - Piperazine-1,4-diium--2,4-dinitrophenolate--water (1/2/2). AB - In the title 1/2/2 adduct, C(4)H(12)N(2)(2+) x 2C(6)H(3)N(2)O(5)(-) x 2H(2)O, the dication lies on a crystallographic inversion centre and the asymmetric unit also has one anion and one water molecule in general positions. The 2,4 dinitrophenolate anions and the water molecules are linked by two O-H...O and two C-H...O hydrogen bonds to form molecular ribbons, which extend along the b direction. The piperazine dication acts as a donor for bifurcated N-H...O hydrogen bonds with the phenolate O atom and with the O atom of the o-nitro group. Six symmetry-related molecular ribbons are linked to a piperazine dication by N---H.O and C---H.O hydrogen bonds. PMID- 11870307 TI - The alpha-D anomer of 5-aza-7-deaza-2'-deoxyguanosine. AB - In the monohydrate of 2-amino-8-(2-deoxy-alpha-D-erythro-pentofuranosyl)-8H imidazo[1,2-a][1,3,5]triazin-4-one, C(10)H(13)N(5)O(4) x H(2)O, denoted (I) or alpha Z(d), the conformation of the N-glycosylic bond is in the high-anti range [chi = 87.5 (3)]. The 2'-deoxyribofuranose moiety adopts a C2'-endo,C3' exo((2')T(3')) sugar puckering (S-type sugar) and the conformation at the C4'-C5' bond is -sc (trans). PMID- 11870306 TI - Ethyl 2-[[2-(3-nitrophenyl)-5-phenyl-1H-imidazol-4-yl]sulfanyl]acetate: synthesis via a microwave-mediated combinatorial chemistry approach. AB - The orange title compound, C(19)H(17)N(3)O(4)S, can be synthesized either via microwave-mediated combinatorial chemistry strategies or conventional synthetic procedures. The phenyl and meta-nitrophenyl C(6) rings are essentially coplanar with the central imidazolyl ring, with interplanar angles of 0.87 (5) and 0.97 (4), respectively, resulting in optimum conjugation (SCH(2) moiety included); lambda(max) = 281 nm in CH(3)CN. The principal intermolecular interactions are N(imid)-H...O(nitro) and N(imid)-H...O[double bond]C [N...O = 3.058 (2) and 3.432 (3) A, and N-H...O = 128 and 153, respectively]. The closest H.S distance is an intramolecular C-H...S contact, with H...S = 2.54 A and C-H...S = 136. PMID- 11870308 TI - [4-(Trifluoromethyl)phenyl]acetonitrile. AB - The crystal structure of the title compound, C(9)H(6)F(3)N, at 123 K contains molecules linked together via several C-H...F and C-H...N contacts, the strongest of which are 2.58 and 2.65 A, respectively. Apparently, an F atom in the CF(3) group is able to compete with a cyano N atom for aromatic H atoms but is less prone to interact with the more acidic methylene H atoms. The Ph-CH(2)CN torsion angle is -6.4 (2) and the planar phenyl ring exhibits a typical deformation of the endo angles at the ipso-C atoms, due to the difference in the electron withdrawing power of the CF(3) and CH(2)CN substituents. PMID- 11870310 TI - Flat versus twisted rotamers of 2,4-disubstituted thiazoles: the effect of intermolecular hydrogen bonds. AB - In the title compounds, 2-amino-4-(2-chloro-4,5-dimethoxyphenyl)-1,3-thiazole, C(11)H(11)ClN(2)O(2)S, (I), and 4-(2-chloro-4,5-dimethoxyphenyl)-2-methyl-1,3 thiazole, C(12)H(12)ClNO(2)S, (II), the dihedral angles between the thiazole moiety and the chloroaryl group are 51.61 (10) and 8.44 (14), respectively. This difference is a consequence of intermolecular hydrogen bonds forcing the stabilization of a twisted rotamer in (I). Substitution of the amino function by a methyl group precludes these contacts, giving a flat rotamer in (II). PMID- 11870309 TI - 1-Methyl-5,6,7,8-tetrahydroquinoxalin-2(1H)-one. AB - The title compound, C(9)H(12)N(2)O, crystallizes in a triclinic unit cell, with two crystallographically independent molecules in the asymmetric unit. The two independent molecules adopt different modes of packing. One type of molecule is arranged in infinite columns, while the other type packs as dimers, forming spacers between the parallel columns. Each type of molecule is arranged in pairs related by inversion centers. The distances between potential reaction centers are 3.395 (2), 3.457 (2) and 3.522 (2) A. As a result of the symmetry of the pairs and the close distances between the potential photoreactive centers, it is expected that the dimer will be the anti-trans isomer. PMID- 11870311 TI - (+/-)-3-Oxocyclohexanecarboxylic and -acetic acids: contrasting hydrogen-bonding patterns in two homologous keto acids. AB - The crystal structures for the title compounds reveal fundamentally different hydrogen-bonding patterns. (+/-)-3-Oxocyclohexanecarboxylic acid, C(7)H(10)O(3), displays acid-to-ketone catemers having a glide relationship for successive components of the hydrogen-bonding chains which advance simultaneously by two cells in a and one in c [O...O = 2.683 (3) A and O-H...O = 166]. A pair of intermolecular close contacts exists involving the acid carbonyl group. The asymmetric unit in (+/-)-3-oxocyclohexaneacetic acid, C(8)H(12)O(3), utilizes only one of two available isoenthalpic conformers and its aggregation involves mutual hydrogen bonding by centrosymmetric carboxyl dimerization [O.O = 2.648 (3) A and O-H...O = 171]. Intermolecular close contacts exist for both the ketone and the acid carbonyl group. PMID- 11870312 TI - 2-Methylphenyl 2-methoxyacridine-9-carboxylate. AB - The title compound, C(22)H(17)NO(3), crystallizes in the monoclinic space group P2(1)/c with four molecules per unit cell. The molecules are arranged in centrosymmetric pairs, joined via the C and attached H atoms in the meta position relative to the methoxy group. These pairs are bonded in the crystalline phase as a result of non-specific dispersive interactions, and through a network of C H...O interactions involving the non-bonded O atom of the carboxy group and, to some extent, the O atom of the methoxy group. The methoxy substituent lies in the plane of the almost planar acridine moiety and is directed towards the phenyl ester group. The phenyl ester group itself is twisted by 35.9 (5) degrees relative to the mean plane of the acridine moiety. PMID- 11870313 TI - trans-3-Ethyl-cis-2,6,6-trimethyl-4-oxocyclohexanecarboxylic acid: an intermediate in the synthesis of a highly potent estrogen. AB - The title compound, C(12)H(20)O(3), (IV), the ethyl ester of which is an intermediate in the synthesis of a compound reported to be highly estrogenic, has been prepared. After the initial steps reported for the synthesis of this ester intermediate were followed, it was converted into the crystalline acid, (IV), for X-ray analysis. It was verified that (IV) was racemic when prepared. X-ray analysis showed that anti-hydrogenation of the double bond had occurred in the synthesis, making the orientation of the carboxyl group cis to the 2-methyl group and trans to the 3-ethyl group. NMR spectroscopy showed that the stereochemistry of (IV) was identical with that of its ester precursor. While the earlier report did not note the stereochemistry of this ester, it pointed out that the estrogenic product derived from it possessed the opposite carboxyl-2-methyl orientation, i.e. trans, although no X-ray analysis was performed. In the light of these results and the importance of correlating biological activity with compound structure, the unequivocal characterization of the highly estrogenic compound is warranted. PMID- 11870315 TI - Three tetrahydroisoquinolinedione derivatives. AB - N-(2-Chlorobenzyl)-1,2,3,4-tetrahydroisoquinoline-1,3-dione, C(16)H(12)ClNO(2), crystallizes in P2(1)/n with three crystallographically independent molecules in the asymmetric unit, which differ slightly in conformation, N-(2-bromo-4 methylphenyl)-1,2,3,4-tetrahydroisoquinoline-1,3-dione, C(16)H(12)BrNO(2), crystallizes in P2(1)/n with one molecule in the asymmetric unit and N-(2,3 dichlorophenyl)-1,2,3,4-tetrahydroisoquinoline-1,3-dione, C(15)H(9)Cl(2)NO(2), crystallizes in P2(1)/c with one molecule in the asymmetric unit. In all three structures, the heterocyclic rings adopt approximately planar conformations. The pyridine rings are orthogonal to the substituted phenyl rings. In all three structures, the crystal packing is stabilized by intermolecular C-H...O hydrogen bonds. PMID- 11870314 TI - 4-(3,17-Dioxoandrost-4-en-16-ylidenemethyl)benzonitrile. AB - The title compound, C(27)H(29)NO(2), has the outer six-membered ring in a sofa conformation, while the central rings are in chair conformations. The five membered ring adopts a slightly distorted 13 beta,14 alpha-half-chair conformation. The cyanobenzylidene moiety has an E configuration with respect to the carbonyl group at position 17. PMID- 11870316 TI - 5-Butyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3-(2,4,6 trimethylphenyl)isoxazole. AB - A novel [3+2]-cycloaddition reaction of alkynylboronates and nitrile oxides gave the title compound, C(22)H(32)BNO(3), as a single regioisomer. The X-ray crystal structure analysis of this compound shows two independent molecules in the asymmetric unit, each with approximately coplanar isoxazole and boronate rings. PMID- 11870317 TI - Steroid 19,B-dinor-8,10-iso-analogues. AB - The Birch reduction of 3-methoxy-B-nor-8-isoestra-1,3,5(10)-trienes followed by acid hydrolysis produces steroid androgen 19,B-dinor-8,10-iso-analogues. By means of X-ray analysis and correlation NMR spectroscopy of 16,16-dimethyl-D-homo-19,B dinor-8-isotestosterone, C(20)H(30)O(2), it is demonstrated that the main conformations in the crystal and in solution for two 19,B-dinor-8,10-iso analogues are, in general, the same. PMID- 11870318 TI - D-Secoestrone derivatives. VI. 17 beta-Benzyl-17 alpha-hydroxy-3-methoxyestra 1,3,5(10)-trien-16-one. AB - The title molecule, C(26)H(30)O(3), shows a novel chemical rearrangement of the substituents at position 17, i.e. an alpha-orientation of the hydroxy group and a beta-orientation of the bulky benzyl moiety. The packing arrangement consists of coils formed by O(2)...O(3) hydrogen bonds along the c axis. The compound shows complete loss of oestrogenic activity, and neither does it exhibit an antagonistic effect. PMID- 11870319 TI - 4-Amino derivatives of 7-nitro-2,1,3-benzoxadiazole: the effect of the amino moiety on the structure of fluorophores. AB - The crystal structures of three 4-amino derivatives of 7-nitro-2,1,3 benzoxadiazole with increasing substituent ring size, viz. 7-nitro-4-(pyrrolidin 1-yl)-2,1,3-benzoxadiazole, C(10)H(10)N(4)O(3), 7-nitro-4-(piperidin-1-yl)-2,1,3 benzoxadiazole, C(11)H(12)N(4)O(3), and 4-(azepan-1-yl)-7-nitro-2,1,3 benzoxadiazole, C(12)H(14)N(4)O(3), have been determined in order to understand their photophysical behaviour. All three were found to crystallize in centrosymmetric space groups. There is considerable electron delocalization compared with the parent compound, although the five-membered oxadiazole ring apparently does not participate in this. The length of the C-N bond between the amino N atom and the 7-nitrobenzoxadiazole system is found to be shorter than in similar compounds, as is the C-N(nitro) bond. In each structure, the nitro group lies in the plane of the benzoxadiazole unit. PMID- 11870320 TI - The diastereoisomers methyl 5-(S)-[2-(R)/(S)-methoxycarbonyl)-2,3,4,5 tetrahydropyrrol-1-ylcarbonyl]-1-(4-methylphenyl)-4,5-dihydropyrazole-3 carboxylate. AB - The title diastereoisomers, methyl 5-(S)-[2-(S)-methoxycarbonyl)-2,3,4,5 tetrahydropyrrol-1-ylcarbonyl]-1-(4-methylphenyl)-4,5-dihydropyrazole-3 carboxylate and methyl 5-(S)-[2-(R)-methoxycarbonyl)-2,3,4,5-tetrahydropyrrol-1 ylcarbonyl]-1-(4-methylphenyl)-4,5-dihydropyrazole-3-carboxylate, both C(19)H(23)N(3)O(5), have been studied in two crystalline forms. The first form, methyl 5-(S)-[2-(S)-methoxycarbonyl)-2,3,4,5-tetrahydropyrrol-1-ylcarbonyl]-1-(4 methylphenyl)-4,5-dihydropyrazole-3-carboxylate--methyl 5-(S)-[2-(R) methoxycarbonyl)-2,3,4,5-tetrahydropyrrol-1-ylcarbonyl]-1-(4-methylphenyl)-4,5 dihydropyrazole-3-carboxylate (1/1), 2(S),5(S)-C(19)H(23)N(3)O(5) x 2(R),5(S) C(19)H(23)N(3)O(5), contains both S,S and S,R isomers, while the second, methyl 5 (S)-[2-(S)-methoxycarbonyl)-2,3,4,5-tetrahydropyrrol-1-ylcarbonyl]-1-(4 methylphenyl)-4,5-dihydropyrazole-3-carboxylate, 2(S),5(S)-C(19)H(23)N(3)O(5), is the pure S,S isomer. The S,S isomers in the two structures show very similar geometries, the maximum difference being about 15 on one torsion angle. The differences between the S,S and S,R isomers, apart from those due to the inversion of one chiral centre, are more remarkable, and are partially due to a possible rotational disorder of the 2-(methoxycarbonyl)tetrahydropyrrole group. PMID- 11870321 TI - The beta-polymorph of phenazine. AB - In beta-phenazine, C(12)H(9)N(2), the molecules show a sandwich herring-bone type of packing. The experimental crystal structure shows very good agreement with that predicted earlier from systematic searches of potential packing arrangements for the known unit cell [Hammond, Roberts, Smith & Docherty (1999). J. Phys. Chem. B, 103, 7762-7770]. PMID- 11870322 TI - Dispiro[2H-benzimidazole-2,1'-cyclohexane-4',2''-[2H]benzimidazole] 1-oxide and dispiro[2H-benzimidazole-2,1'-cyclohexane-4',2''-[2H]benzimidazole] 1,1'' dioxide. AB - Oxidation of tetrahydrodispirobenzimidazole by m-chloroperbenzoic acid did not produce dispiro-2H-benzimidazole, which is the product obtained by oxidation with MnO(2). Instead, a mixture of two compounds was identified, namely dispiro[2H benzimidazole-2,1'-cyclohexane-4',2''-[2H]benzimidazole] 1-oxide, C(18)H(16)N(4)O, (III), and dispiro[2H-benzimidazole-2,1'-cyclohexane-4',2'' [2H]benzimidazole] 1,1''-dioxide, C(18)H(16)N(4)O(2), (IV). In (III), the molecules are disordered about a twofold rotation axis and have 2/m site symmetry. In (IV), the crystals are triclinic and the molecules occupy crystallographic inversion centers. Although the two compounds are very similar and are arranged in layers, they adopt completely different packing modes within the layers, viz. herring-bone in (III) and parallel molecules in (IV). The molecules within the layers are held together by C-H...O and C-H...N hydrogen bonds. PMID- 11870323 TI - Cyclohexylphosphonic Acid. AB - The title compound, C(6)H(13)O(3)P, displays a crystallographic mirror plane. Bond lengths in the phosphonic acid moiety are P[double bond]O = 1.5557 (13) A and P[double bond]O = 1.5089 (18) A. The molecules are linked via intermolecular hydrogen bonding to form a one-dimensional chain of fused rings. There are no significant contacts between planes. PMID- 11870324 TI - Tetrakis(chloromethyl)phosphonium chloride monohydrate. AB - Tetrakis(chloromethyl)phosphonium chloride monohydrate, C(4)H(8)Cl(4)P(+) x Cl(-) x H(2)O or P(CH(2)Cl)(4)(+) x Cl(-) x H(2)O, is the first crystal structure determination of a tetrakis(halogenomethyl)phosphonium compound to date. The only comparable structures known so far are of phosphonium ions containing just one halogenomethyl group. The solvent water molecule interacts with the Cl(-) anion via hydrogen bonds, with O...Cl distances of 3.230 (2) and 3.309 (2) A. The structure also contains several C-H...Cl(-) and C-H...O contacts, though with longer D...A distances [D.A 3.286 (3)-3.662 (2) A] or bent D-H...A angles. For these reasons, the C-H...Cl(-) and C-H...O interactions should not be considered as strong hydrogen bonds. PMID- 11870325 TI - 1,3-Dimethyl-2-oxo-4,6-diphenyl-1,2,3,4-tetrahydropyridine-3-carbonitrile. AB - The crystal structure of the title compound, C(20)H(18)N(2)O, reveals a distorted half-chair conformation of the central tetrahydropyridine (THP) ring, with the cyano- and adjacent phenyl-substituted C atoms displaced by 0.329 (1) and -0.315 (1) A, respectively, from the THP best plane. Steric interactions force the phenyl rings out of the THP plane by 49.21 (9) and 65.76 (5). The cyano moiety is coplanar with the THP plane. PMID- 11870326 TI - 1,3-Bis(2,4-dibromophenyl)triazene. AB - The crystal structure of the title compound, C(12)H(7)Br(4)N(3), shows that the stereochemistry about the N[double bond]N double bond of the N[double bond]N-N(H) moiety is trans. The whole molecule deviates slightly from planarity (r.m.s. deviation 0.164 A). While one of the aryl substituents is almost coplanar with the triazene chain, weak intermolecular Br...C contacts cause the second aryl substituent to deviate by an angle of 9.1 (8) from the plane defined by the N[double bond]N-N group. Weak intermolecular N-H...Br interactions between molecules related by the diagonal glide plane give rise to chains, which are stacked along the [100] crystallographic direction. An unequal distribution of double-bond character between the N atoms suggests a delocalization of pi electrons over the diazoamino group and the adjacent aryl groups. PMID- 11870327 TI - Three ginkgolide hydrates from Ginkgo biloba L.: ginkgolide A monohydrate, ginkgolide C sesquihydrate and ginkgolide J dihydrate, all determined at 120 K. AB - A low-temperature structure of ginkgolide A monohydrate, (1R,3S,3aS,4R,6aR,7aR,7bR,8S,10aS,11aS)-3-(1,1-dimethylethyl)-hexahydro-4,7b dihydroxy-8-methyl-9H-1,7a-epoxymethano-1H,6aH-cyclopenta[c]furo[2,3 b]furo[3',2':3,4]cyclopenta[1,2-d]furan-5,9,12(4H)-trione monohydrate, C(20)H(24)O(9) x H(2)O, obtained from Mo K alpha data, is a factor of three more precise than the previous room-temperature determination. A refinement of the ginkgolide A monohydrate structure with Cu K alpha data has allowed the assignment of the absolute configuration of the series of compounds. Ginkgolide C sesquihydrate, (1S,2R,3S,3aS,4R,6aR,7aR,7bR,8S,10aS,11S,11aR)-3-(1,1 dimethylethyl)-hexahydro-2,4,7b,11-tetrahydroxy-8-methyl-9H-1,7a-epoxymethano 1H,6aH-cyclopenta[c]furo[2,3-b]furo[3',2':3,4]cyclopenta[1,2-d]furan-5,9,12(4H) trione sesquihydrate, C(20)H(24)O(11) x 1.5H(2)O, has two independent diterpene molecules, both of which exhibit intramolecular hydrogen bonding between OH groups. Ginkgolide J dihydrate, (1S,2R,3S,3aS,4R,6aR,7aR,7bR,8S,10aS,11aS)-3-(1,1 dimethylethyl)-hexahydro-2,4,7b-trihydroxy-8-methyl-9H-1,7a-epoxymethano-1H,6aH cyclopenta[c]furo[2,3-b]furo[3',2':3,4]cyclopenta[1,2-d]furan-5,9,12(4H)-trione dihydrate, C(20)H(24)O(10) x 2H(2)O, has the same basic skeleton as the other ginkgolides, with its three OH groups having the same configurations as those in ginkgolide C. The conformations of the six five-membered rings are quite similar across ginkgolides A-C and J, except for the A and F rings of ginkgolide A. PMID- 11870328 TI - The anti-inflammatory triterpenoid methyl 2-cyano-3,12-dioxooleana-1,9(11)-dien 28-oate methanol solvate hydrate. AB - The title compound, C(32)H(43)NO(4) x CH(4)O x H(2)O, has a nearly planar cyano enone A ring in an otherwise normal oleanane triterpenoid. Rings A, B and C are non-chairs, but rings D and E adopt essentially cyclohexane chair conformations. The structure clearly establishes the C-D-E ring stereochemistry as trans-syn cis, as predicted from a nuclear Overhauser effect (NOE) NMR measurement. PMID- 11870329 TI - Clinical aspects of autoimmune pancreatitis in Sjogren's syndrome. PMID- 11870330 TI - Radical distal pancreatectomy with en bloc resection of the celiac artery, plexus, and ganglions for advanced cancer of the pancreatic body: a preliminary report on perfect pain relief. AB - OBJECTIVE: The purpose of this study was to report the effect of radical distal pancreatectomy with en bloc resection of the celiac artery, plexus, and ganglions for locally advanced cancer of the pancreatic body on intractable abdominal and/or back pain and to explore the histopathologic mechanism of this pain. PATIENTS: Five patients with pancreatic body cancer involving the celiac and/or common hepatic artery underwent this radical surgery intended to cure the cancer. DESIGN: A retrospective analysis was performed. MAIN OUTCOME MEASURES: Surgical magnitude, postoperative pain control, postoperative outcome, and histopathologic findings were studied. RESULTS: Arterial reconstruction, gastrointestinal reconstruction, and blood transfusions were unnecessary. The organ deficit was limited to the distal pancreas, spleen and left adrenal gland. There was no postoperative mortality. Postoperative complications occurred in four patients, who were successfully managed with medical treatment. This led to prolonged hospital stays. The intractable preoperative abdominal and/or back pain was completely relieved immediately after surgery in all patients. Perfect pain control has been maintained from surgery to the last follow-up. Histopathologic examination of the surgical specimens revealed cancer invasion of the celiac plexus in all patients. CONCLUSIONS: This operation offers not only disease radicality but also perfect pain relief. The survival benefit has not yet been fully defined. PMID- 11870331 TI - The impact on clinical practice of endoscopic ultrasonography used for the diagnosis and staging of pancreatic adenocarcinoma. AB - CONTEXT: Endoscopic ultrasonography is considered a highly accurate procedure for diagnosing small pancreatic tumors and assessing their locoregional extension. OBJECTIVE: To evaluate the impact of endoscopic ultrasonography on the management of pancreatic adenocarcinoma in clinical practice. PATIENTS: Sixty-four consecutive patients (mean age 70.5 plus/minus 11.9 years) hospitalized for staging or diagnosis of pancreatic adenocarcinoma were retrospectively (from January 1995 to November 1997) or prospectively studied (from December 1997 to August 1999). SETTING: Group 1 consisted of 52 patients with pancreatic adenocarcinoma which was discovered using computerized tomography scanning and/or ultrasound. Endoscopic ultrasonography was utilized for staging purposes only in patients who were considered to be operable and the tumor to be resectable based on computerized tomography scanning criteria. Group 2 consisted of 12 patients who were diagnosed as having a pancreatic adenocarcinoma using endoscopic ultrasonography whereas computerized tomography scanning and ultrasound was negative. MAIN OUTCOME MEASURES: The impact of endoscopic ultrasonography was analyzed on the basis of the number of patients requiring endoscopic ultrasonography as a staging procedure (Group 1) and by evaluating the performance of endoscopic ultrasonography in determining resectability (Groups 1 and 2) based on the surgical and anatomopathological results. RESULTS: Endoscopic ultrasonography was performed in 20 out of 64 patients (31.3%): 8/52 in Group 1 (15.4%) and all 12 patients of Group 2. Endoscopic ultrasonography correctly assessed an absolute contraindication to resection in 11 cases. Resection was confirmed in 8 of the 9 cases selected by endoscopic ultrasonography. The positive predictive value, negative predictive value and overall accuracy of endoscopic ultrasonography for determining resection were 89%, 100%, and 95%, respectively. CONCLUSIONS: The impact of endoscopic ultrasonography seems especially relevant for the detection of pancreatic tumors after negative computerized tomography scanning, and for the prevention of unnecessary laparotomies as complementary staging after ultrasonography and computerized tomography scanning. PMID- 11870332 TI - Neurotrophins and neurotrophin receptors mRNAs expression in pancreatic islets and insulinoma cell lines. AB - CONTEXT: It is worth noting that islets and betaTC6-F7 cells share a common pattern of expression of neurotrophins and neurotrophin receptors. Recently, several studies have hypothesized a role for nerve growth factor in pancreatic development and maturation, suggesting that nerve growth factor may be a survival factor for pancreatic beta-cells. OBJECTIVE: The aim of the present study was to investigate the pattern of expression of neurotrophins and their relative receptors both in rat pancreatic islets and in a wide panel of insulinoma cell lines. MAIN OUTCOME MEASURES: A semi-quantitative reverse-transcription polymerase chain reaction analysis was performed on ribonucleic acids extracted from these cells. RESULTS: Reverse transcription-polymerase chain reaction analysis demonstrates that brain-derived neurotrophic factor, as well as neurotrophins 3 and 4, are expressed both in islets and in all insulinoma cells, while nerve growth factor is expressed only in islets, betaTC6-F7 cells and, at a low level, in RIN 1046-38 cells. Receptors protein tyrosine kinase A and C are ubiquitously expressed both in islets and insulinoma cells. Tyrosine kinase B is absent in HIT-T15 cells. CONCLUSIONS: These data indicate that betaTC6-F7 cells are a suitable model for studying the role of neurotrophins in the survival of beta-cells. PMID- 11870333 TI - Jejunal feeding in chronic pancreatitis with severe necrosis. AB - CONTEXT: Necrotizing pancreatitis is the most serious form of pancreatic inflammatory disease leading to multiorgan failure and a high (15-20%) mortality rate. The poor nutritional and metabolic conditions and secondary bacterial translocation raise the mortality rate even more. OBJECTIVE: The aim of the study was to evaluate the effect of jejunal feeding in cases of chronic pancreatitis with extended necrosis. PATIENTS: In our institution, over a five-year period, 86 patients with severe necrotizing pancreatitis were treated for extended necrosis. In 19 patients, chronic calcifying pancreatitis was demonstrated by computed tomography showing more than 20% necrosis in the residual pancreas as well. SETTING: In 12 cases, nutrition was provided by jejunal feeding using an endoscopically placed nasojejunal feeding tube, whereas in 7 cases, hypocaloric parenteral nutrition was used. DESIGN: Retrospective unicenter study. MAIN OUTCOME MEASURES: The rate of healing with conservative treatment. RESULTS: Two of the 12 jejunally fed patients were operated on because of complications of pancreatitis. Five patients required intervention in the hypocaloric parenteral nutrition group: 4 were operated on and one more needed endoscopic intervention. The healing rate was significantly higher (P=0.045) in the jejunal feeding group (83.3%) than in the parenteral nutrition (28.6%) patients. CONCLUSIONS: In cases of chronic calcifying pancreatitis serious necrosis can develop in the residual pancreas resulting in a severe acute pancreatitis-like disease. A better healing rate was achieved and less interventions became necessary using nasojejunal tube feeding than in the parenteral nutrition group and this was analogous to what was observed in severe necrotizing pancreatitis This form of pancreatitis has not yet been described in the literature in detail. The authors suggest that it be regarded as a separate entity. PMID- 11870334 TI - Frontal and sagittal balance analysis of late onset idiopathic scoliosis treated with third generation instrumentation. AB - As scoliotic curve is a rotational deformity, derotation maneuvre was used as the corrective factor, but recent studies demonstrated spinal imbalance and decompensation problems in patients treated with this method. This study evaluates 217 late onset idiopathic scoliosis patients surgically treated with third generation instrumentation (Texas Scotish Rite Hospital System - TSRH) from September 1991 to November 1996 with a minimum 2 years follow up. Preoperative and postoperative Cobb angles in the frontal plane and thoracic kyphosis and lumbar lordosis angles in the sagittal plane are measured. The balance was analyzed clinically and radiologically by measurement of the lateral trunk shift (LT), shift of head (SH) and shift of stable vertebra (SS) in vertebral unit (VU). At final follow - up correction loss, infection and other complications were documented. Mean age of the patients was 14.8 +/- 2.3 and mean follow up period 55.8 +/- 29.5 months. When all the patients were included, preoperative mean Cobb angles of major curves in the frontal plane was 59.1 +/- 20.7 degrees. Major curves that were corrected by 34.8 +/- 20.5 % in the bending radiograms were achieved by 58.9 +/- 19.5 % correction postoperatively. At the last control, 7.3 degrees +/- 6.4 degrees of correction loss was recorded in major curves in the frontal plane. Also postoperative kyphosis angle and lumbar lordosis angles were 31.4 degrees +/- 11.6 degrees and 30.6 degrees +/- 10.9 degrees respectively. Postoperatively, a statistically significant correction was obtained in LT, SH and SS values. None of the patients had complete balance (SH: 0 VU, SS: 0 VU) preoperatively. Only 39.2 % of the patients had clinically balanced curves (0 VU < SH < 0.5 VU and 0 VU < SS < 0.5 VU). Postoperatively, 47.9 % of the patients were found to be completely balanced, while 43.8 % had a balanced curve. Overall 91.7 % of the patients had a trunk balance after surgical intervention. The remaining 8.3 % imbalanced curve rate raised up to 16.6 % at final follow up, but the loss of correction rates in S S and SH values were found to be insignificant. The postoperative "imbalance" problem was mostly seen in Type II and Type IV curves. However, at final follow up, the imbalance problem due to overcorrection which became evident especially by "shift of head" to opposite side was seen in all types of curves. It is established that high correction rates can be obtained in scoliotic curves with third - generation instrumentation. No undue effects were observed in the uninstrumented lumbar curves. Thoracic sagittal contours of the hypokyphotic patients were improved. Use of this instrumentation system causes minimal imbalance problems and with proper preoperative planning high correction rates can be achieved. PMID- 11870335 TI - Methylenetetrahydrofolate reductase gene polymorphism and ischemic stroke: sex difference in Japanese. AB - Moderately elevated plasma homocysteine levels have been established as independent risk factors in vascular disease, including ischemic stroke. Recently, a common mutation (C677T) in the 5,10-methylenetetrahydrofolate reductase (MTHFR) gene reducing the activity of MTHFR and increasing homocysteine levels in plasma was reported. The C677T MTHFR mutation may be a risk factor for ischemic stroke, but the results of previous studies have been conflicting. One possible explanation is that the association with the MTHFR genotype may be different according to gender. To investigate the association for ischemic stroke, we conducted a case-control study of 77 hospital cases (49 men and 28 women) with ischemic stroke and 229 (120 men and 109 women) control subjects in Japanese. The prevalence of conventional vascular risk factors and MTHFR genotypes were determined in case and controls. After adjustment by multiple analysis in all there was no statistical significance in MTHFR genotypes. The conventional vascular risk factors such as diabetes mellitus (adjusted odds ratio [OR], 17.21), hypertension (adjusted OR, 4.67), smoking habit (adjusted OR, 4.70), and hyperlipidemia (adjusted OR, 2.73) were identified independently associated with ischemic stroke. With a separate sex analysis it was identified that the relationship of the MTHFR T/T gneotype was statisticaly significant in women (adjusted OR, 9.49; 95% CI, 1.75-51.47, P=0.0091). The relevance of the MTHFR T/T mutation appears to be restricted to women, suggesting a role of female hormones in the resistance to elevated homocysteine levels due to the MTHFR T/T mutation. PMID- 11870336 TI - Assessment of safety and feasibility of spinal endoscope in the thoracic and lumbar region: a cadaveric study. AB - BACKGROUND: Endoscope has been used far less in the spinal region than in the intracranial cavity. One of the reasons is its safety and feasibility has not been established. To evaluate its safety and feasibility, we performed preliminary cadaveric study prior to clinical endoscopic intervention in the thoracic and lumbar spinal canal. METHODS: The endoscope that had a directable tip with external diameters of 2.2mm was used in this study. The endoscope was inserted percutaneously in the lumbar region (lumbar puncture method) in nine cadavers and was advanced rostrally under endoscopic monitoring. In advancing the endoscope to the upper thoracic region, dorsal, lateral and ventral route was applied. When the endoscope was advanced to the upper thoracic region, vertebral canal from Th 1 to L4 was opened to examine whether there were any injuries to spinal cord and nerves. RESULTS: The endoscope could be manipulated and advanced under endoscopic image in the lumbar region. However, the filum terminale could not be detected under endoscopic view. In the thoracic region, the endoscope could be advanced in dorsal and lateral route. However, in advancing the endoscope in ventral route, the endoscope could not be advanced more rostrally. In cadavers that the endoscope was advanced in dorsal and lateral route, macroscopic and microscopic investigation revealed no injuries including compression marks or trace of cord were observed on the surface of the spinal cord and nerves. CONCLUSIONS: From this preliminary cadaver study, the safe route to advance the endoscope from the lumbar to the thoracic region was established and possibilities of clinical interventions in safe methods could be suggested. PMID- 11870338 TI - Facial nerve monitoring in chronic ear surgery: US practice patterns. AB - OBJECTIVE: The use of intraoperative facial nerve monitoring (IOFNM) improves facial nerve outcomes in acoustic neuroma surgery, but the role of IOFNM in chronic ear surgery (CES) is poorly defined. This study was performed to identify IOFNM practice patterns in the United States for CES. METHODS: A 10-item survey was mailed to a random sample of 500 active members of the American Academy of Otolaryngology-Head and Neck Surgery. RESULTS: The overall response rate was 45.2%. Of the respondents, 75% had access to IOFNM, and of this group, 66% used IOFNM at least some of the time. Otolaryngologists trained in the 1990s (P < 0.0001), those in an academic setting (P = 0.02), and those who perform more otologic than other types of surgery (P = 0.004) were more likely to use IOFNM. Although most respondents had access to IOFNM, only 32% thought that IOFNM should be required for CES. CONCLUSIONS: The majority of otolaryngologists who perform CES have access to IOFNM and will use it at least some of the time. IOFNM use is especially prevalent in otolaryngologists who have trained recently and in more experienced otologic surgeons. However, there are strong feelings against requiring IOFNM for CES. PMID- 11870337 TI - Phase III clinical trial results with the Vibrant Soundbridge implantable middle ear hearing device: a prospective controlled multicenter study. AB - OBJECTIVES: The goal of the study was to evaluate the performance of a semi implantable middle ear hearing device (Vibrant Soundbridge System [VSB]; Symphonix Devices, Inc). STUDY DESIGN: A prospective, single-subject, repeated measures multicenter study was conducted to determine the safety and efficacy of the VSB using analog and digital external processors. Measures included residual hearing, functional gain, speech recognition, acoustic feedback, occlusion, and patient self-assessment to determine satisfaction, perceived performance, and device preference compared with an appropriately fit acoustic hearing aid. Fifty three adult subjects with moderate to severe sensorineural hearing loss were evaluated at 4 or more intervals after implantation. RESULTS: Improvements in satisfaction, performance, and preference were statistically significant with the VSB, as was functional gain across all test frequencies (P < 0.001). Occlusion and feedback were virtually eliminated. Aided speech recognition was comparable between VSB and the hearing aid. Residual hearing was unchanged. CONCLUSION: The VSB is a safe and effective treatment option for adults with moderate to severe sensorineural hearing loss. PMID- 11870339 TI - Distortion-product otoacoustic emissions in nonacoustic tumors of the cerebellopontine angle. AB - OBJECTIVES: The goal of the present study was to compare the frequencies of cochlear and non-cochlear patterns of distortion-product otoacoustic emissions (DPOAEs) in patients with nonacoustic (non-AN) tumors of the cerebellopontine angle (CPA)/internal auditory canal with those in patients with acoustic neuroma (AN). STUDY DESIGN AND SETTING: We conducted a prospective study of patients with non-AN internal auditory canal/CPA tumors treated between 1992 and 1999 in a tertiary care setting. We collected data on behavioral pure tone average, speech discrimination score, acoustic immittance tests, DPOAEs, and tumor size. DPOAEs were divided into those with a cochlear or a noncochlear pattern. Comparisons were made with a control group of patients with AN of the CPA. RESULTS: Twenty four patients were included in the study: 12 patients in the non-AN group and 12 patients in the AN group. In the non-AN group, 5 (42%) patients had a cochlear pattern and 7 (58%) had a noncochlear pattern. In the AN group, 7 (58%) patients had a cochlear pattern and 5 (42%) had a noncochlear pattern. The differences between the non-AN and AN groups were statistically significant. In the non-AN group, patients with a cochlear-pattern DPOAE had significantly smaller tumors (P = 0.03) and a trend (P = 0.07) toward higher speech discrimination score. Age and pure tone average were not significantly different in the non-AN tumor patients. In the AN group, none of the study parameters were significantly related to the pattern of DPOAE. CONCLUSIONS: ANs appear to differ from other tumors of the CPA in their propensity to cause sensory versus neural hearing loss. A larger series of patients would be useful to confirm these preliminary findings. PMID- 11870340 TI - Does the addition of hyperbaric oxygen therapy to the conventional treatment modalities influence the outcome of sudden deafness? AB - OBJECTIVE: To investigate the therapeutic effects of the addition of hyperbaric oxygen (HBO) therapy to the conventional therapies in sudden deafness (SD) and to investigate the influence of patient age on the effectiveness of HBO therapy. STUDY DESIGN AND SETTING: We undertook a retrospective review of 50 cases of SD treated at a tertiary university hospital. Twenty-five patients (group 1) were treated with betahistine hydrochloride, prednisone, and daily stellate ganglion block. A second group (group 2) of 25 patients received the same basic treatment with the addition of HBO therapy. RESULTS: The mean hearing gain was 20.0 dB in group 1 and 37.9 dB in group 2 (P < 0.05). In group 2 patients, the mean gains were 51.4 and 23.3 dB for those younger and older than 50 years (P < 0.05) and 48.9 and 14.5 dB for those younger and older than 60 years (P < 0.001), respectively. In patients older than 60 years, the mean gains were 14.5 and 14.4 dB in group 2 and group 1, respectively (P > 0.05). CONCLUSIONS: The addition of HBO therapy to the conventional treatment significantly improves the outcome of SD, especially in patients younger than 50 years. Additional HBO therapy provides limited benefit in patients older than 50 years and no benefit in patients older than 60 years. PMID- 11870341 TI - The otologic consequences of the Omagh bomb disaster. AB - OBJECTIVES: The goal of the study was to compare the spontaneous healing rate for traumatic perforation of the tympanic membrane sustained in the Omagh bomb disaster with that of previous reports in the literature. DESIGN AND SETTING: A retrospective study was conducted at Tyrone County Hospital, Northern Ireland, 2 years after the incident. PARTICIPANTS: One hundred thirty-eight patients who sustained otologic injuries from the bomb blast were included in the study. RESULTS: Only 47 of the 124 perforations healed spontaneously. The previously reported spontaneous healing rate for traumatic perforations of the tympanic membrane is 80% to 90%; in our study, the healing rate was unexpectedly low at 38%. CONCLUSION: The massive explosion in a narrow street with hundreds of persons in close proximity to the bomb may account for the poor healing rate. PMID- 11870342 TI - The oto-palato-digital syndrome: variable clinical expressions. AB - The oto-palato-digital (OPD) syndrome is a rare but well-defined disorder characterized by a skeletal dysplasia of the hands and feet, hearing loss, and anomalies of the palate. Since it was first described by Taybi in 1962, a little over 30 cases have been reported in the literature. A more lethal variant of the syndrome was described later by Fitch and was termed OPD type II. Several studies were conducted to determine the exact mode of inheritance of this syndrome, whereas others have focused on the characterization of the skeletal anomalies and their variations. Otologists were interested in determining the etiology of the associated hearing loss. We report 4 cases of patients with the spectrum of anomalies that characterize the OPD syndrome. These patients include 3 siblings and 1 unrelated patient who presented to our service complaining of hearing loss. The skeletal anomalies, special features, and audiologic findings are described and compared with those in previously reported cases. A discussion based on a literature review of the mode of inheritance, of variation in the clinical expression, and of the etiology of hearing loss is also included. Finally, we review and discuss the subdivision of this syndrome into the 2 types (I and II). PMID- 11870343 TI - Aspirin-sensitive versus non-aspirin-sensitive nasal polyp patients: analysis of leukotrienes/Fas and Fas-ligand expression. AB - OBJECTIVE: The goal of the present study was to identify levels of leukotrienes released by leukocytes and nasal polyps from aspirin-sensitive patients (ASPs) and non-aspirin-sensitive patients (NASPs) after exposure to various concentrations of aspirin. Eosinophil expressions of apoptosis regulators were also compared. STUDY DESIGN AND SETTING: Leukotriene release from nasal polyps and leukocytes of ASPs and NASPs was measured after exposure to differing concentrations of aspirin. Eosinophils were analyzed for Fas and Fas ligand receptors using flow cytometry. RESULTS: NASPs showed increased leukotriene release from nasal polyps on increased aspirin exposure. Polyps from ASPs did not show any change. ASPs showed greater leukotriene release from blood on increasing aspirin exposure compared with NASPs. Flow cytometric analysis showed eosinophils in the peripheral blood of ASPs have decreased Fas expression compared with those of NASPs. CONCLUSIONS: This study showed differences in leukotriene release and eosinophil apoptosis between ASPs and NASPs. SIGNIFICANCE: These findings suggest a role of leukotrienes and eosinophils in aspirin sensitivity mechanisms in blood. PMID- 11870344 TI - Management of sphenoid sinus cerebrospinal fluid rhinorrhea: making use of an extended approach to the sphenoid sinus. AB - OBJECTIVES: Specific information addressing the management of cerebrospinal fluid (CSF) fistulas that originate from within the sphenoid sinus remains scant. The objective of this study was to review the cause and management of CSF rhinorrhea arising from the sphenoid sinus. STUDY DESIGN AND SETTING: This is a retrospective chart review of 12 cases of CSF rhinorrhea arising from the sphenoid sinus that occurred in 11 patients. All patients were treated at a single institution between 1994 and 1999. RESULTS: All patients were managed surgically with sphenoid sinus fat obliteration using an endoscopic sublabial, transseptal approach. This approach was successful for all 12 cases, with median duration of follow-up of 18 months. CONCLUSIONS: Endoscopically assisted transseptal repair of CSF fistulas that originate within the sphenoid sinus offers an alternate approach to previously described methods of repair in this region. Advantages include wide access to the entire sphenoid sinus, improved access to laterally pneumatized regions within the sphenoid sinus, and rostral mucosal closure over the repair within the sinus. PMID- 11870345 TI - Outcome predictors for endoscopic sinus surgery. AB - OBJECTIVE: The outcomes management of endoscopic sinus surgery (ESS) is a major effort to improve the quality of chronic sinusitis treatment. The objective of this study was to investigate the factors that may predict a variety of outcomes of ESS. STUDY DESIGN AND SETTING: Two hundred and thirty consecutive patients who underwent ESS were prospectively evaluated by computerized axial tomography scan of the sinus and a validated chronic sinusitis survey (CSS) before surgery. The effects of prognostic factors were then investigated. RESULTS: Extent of disease was a consistent predictor (P < 0.05) for bleeding, complication occurrence, medical resource utilization, subjective sinus-specific health status, and physicians' objective evaluation of surgical outcomes. The preoperative CSS total score and extent of disease significantly (P < 0.05) predicted the postoperative CSS total score. Subjective and objective outcomes were significantly correlated (r = 0.58, P = 0.0001). These findings suggested the necessity of disease severity stratification when reporting ESS outcomes. CONCLUSION: Operation related events, medical resource utilization, physician evaluation, and sinus specific health status of patients are all integral parts of reporting outcomes of ESS. We have concluded that computerized axial tomography scan of the sinuses and CSS are efficient ways of providing reliable baseline information before ESS. PMID- 11870346 TI - The effects of an external nasal dilator and nasal dimensions in Asians. AB - OBJECTIVES: We examined the potential correlation between the effectiveness of an external nasal dilator for Asians and nasal dimensions. STUDY DESIGN AND SETTINGS: The length, width, and height of the subjects' nose were measured. From these parameters, a nasal index (nasal width/nasal length) and a basal view index (columella height/nasal width) were calculated. Nasal airway resistances with and without an external nasal dilator were measured. RESULTS: There was a significant correlation between both nasal index and basal view index and the changes in nasal resistance that resulted from wearing an external nasal dilator, whereas no significant correlation was detected between each of the 3 nasal dimensions and changes in nasal resistance. CONCLUSIONS: Noses with a smaller nasal index or a larger basal view index receive greater benefits from the wearing of an external nasal dilator. The effects of external nasal dilators are influenced by nasal shape rather than by individual nasal dimensions in Asians. PMID- 11870347 TI - Lung volume dependence of pharyngeal cross-sectional area by acoustic pharyngometry. AB - Pharyngeal size and the dynamic behavior of the pharynx may be important factors in the modulation of pharyngeal airflow. There are two measures of pharyngeal function: changes in pharyngeal area with lung volume and changes in pharyngeal area in response to externally applied positive pressure. Both measurements have been used for the assessment of pharyngeal function, and both reflect pharyngeal "floppiness." The aim of the present study was to examine the relationship between pharyngeal cross-sectional area, using acoustic reflection technique, and different lung volumes (tidal volume, inspiratory reserve volume, and expiratory reserve volume), to determine whether there are differences in mechanical properties of the pharynx of normal volunteers in response to changes in intrapharyngeal pressure. The acoustic technique was used to assess the pharyngeal cross-sectional area of 40 healthy volunteers (29 men and 11 women) at tidal volume, inspiratory reserve volume, and expiratory reserve volume. In men, the mean pharyngeal area at tidal volume was 3.191 cm2, the mean pharyngeal area at inspiratory reserve volume was 2.976 cm2, and the mean pharyngeal area at expiratory reserve volume was 2.975 cm2. In women, the corresponding pharyngeal area measurements were 2.832, 2.484, and 2.492 cm2. Statistical analysis of results showed that men have a larger pharyngeal cross-sectional area and the pharyngeal airways of men and women act in a similar manner in response to changes in intrapharyngeal pressure, with men having a greater change. Examination of the pharyngeal compliance by acoustic pharyngometry adds to the potential of this technique as a tool for the evaluation of the pharyngeal airway in terms of area and dynamic behavior assessment. This may be of relevance in promotion of the development of upper airway assessment in patients with obstructive sleep apnea. PMID- 11870348 TI - Objective and early diagnosis of chylous fistula in the postoperative period. AB - OBJECTIVE: The study goal was to evaluate whether quantitative analysis of neck drainage contents is an important parameter of objective and early diagnosis of chylous fistula (CF) in the postoperative period. STUDY DESIGN AND SETTING: We prospectively evaluated 103 consecutive neck dissections during a 2-year period at a tertiary academic referral center. Six of the patients (5.8%) had CF. The data obtained from the patients with CF were matched with normal data obtained from patients without CF. RESULTS: A statistically significant difference was found between total drainage volumes of the patients with and those without CF (P < 0.05). In almost all patients with CF, drainage levels of triglycerides and cholesterol on postoperative day 1 were higher than serum levels. The difference between the drainage levels of triglycerides and cholesterol of the patients with and without CF was highly significant (P < 0.001). CONCLUSIONS: Serum and drainage levels of triglycerides and cholesterol on postoperative day 1 can be used as objective and early predictive parameters of CF. PMID- 11870349 TI - Tuberculous and nontuberculous cervical lymphadenitis: a clinical review. AB - OBJECTIVES: The aim of the present study was to identify differences in clinical characteristics between patients with tuberculous cervical lymphadenitis and those with nontuberculous cervical lymphadenitis and to determine the diagnostic accuracy of fine needle aspiration (FNA) cytology. STUDY DESIGN AND SETTING: Seventy-two patients with inflammatory cervical lymphadenitis were studied retrospectively. They were divided into 2 groups: group 1 consisted of those with tuberculous lymphadenitis and group 2 consisted of those with non-tuberculous lymphadenitis. The demographic characteristics, clinical parameters, and hematological and cytological results of the 2 groups were compared. RESULTS: Other than there being a significantly higher proportion of foreign-born patients in group 1, there were no differences in clinical characteristics between the 2 groups. The sensitivity and specificity of FNA cytology in the diagnosis of tuberculous lymphadenitis were 88% and 96%, respectively. CONCLUSION: It is difficult to clinically differentiate tuberculous from nontuberculous lymphadenitis. FNA cytology is useful in the diagnosis of tuberculous lymphadenitis. SIGNIFICANCE: In regions where tuberculosis is endemic, treatment can be instituted without the need for excisional biopsy if the FNA results show characteristic caseating granuloma. PMID- 11870350 TI - Reassessment of CD62L as a marker of pre-effector T cells in the tumor draining lymph nodes of head and neck cancer patients. AB - OBJECTIVES: CD62L was evaluated as a determinant of human pre-effector T cells. STUDY DESIGN AND SETTING: Phenotype and cytokine secretion profiles of CD62L cells were determined based on activation status. RESULTS: CD62L(Low) T cells demonstrated significantly higher secretion of interleukin (IL)-10 and interferon (IFN)-gamma than did CD62L(High) T cells. After activation, the majority of cells expressed high levels of the CD62L surface marker. Postactivation levels of IL-10 production remained elevated or unchanged. In a murine B16 melanoma model, freshly isolated CD62L(Low) tumor draining lymph nodes (TDLN) T cells showed increased secretion of IL-2 and IL-4 but not of IL-10 or IFN-gamma. The surface expression of CD62L and cytokine secretion patterns were maintained after activation with concomitant increases in IL-10. CONCLUSION: Our results provide evidence that CD62L(Low) T cells in TDLNs of progressively growing squamous cell carcinoma of the head and neck differ phenotypically and functionally from those of mouse origin. SIGNIFICANCE: Characterization of this human CD62L(Low) T cell population provides initial insight regarding novel surface markers in TDLN T cells that might correlate with antitumor reactivity. PMID- 11870351 TI - A unique combination of pre- and intraoperative modalities for the diagnosis and localization of recurrent medullary thyroid carcinoma. PMID- 11870352 TI - Madelung's lipomatosis associated with head and neck malignant neoplasia: a study of 2 cases. PMID- 11870353 TI - Sternoclavicular joint hypertrophy after neck dissection and upper trapezius myocutaneous flap transposition. PMID- 11870354 TI - Radiation therapy as an option for upper airway obstruction due to Wegener's granulomatosis. PMID- 11870355 TI - Malignant neoplasia at different ages presenting as peritonsillar abscess. PMID- 11870356 TI - Sebaceous adenoma in the submandibular gland. PMID- 11870357 TI - Primary bilateral tuberculous dacryocystitis with preauricular lymphadenopathy: a diagnostic difficulty of recent times. PMID- 11870358 TI - A case of invasive aspergillosis of the paranasal sinuses with a feature of allergic Aspergillus sinusitis. PMID- 11870359 TI - Ossicular chain dystopia and dilation of vestibules in branchio-oto-renal syndrome: a new phenotypic association? PMID- 11870360 TI - Sheila: Professor Dame Sheila Sherlock (1918-2001). PMID- 11870362 TI - Gene repair in the new age of gene therapy. PMID- 11870363 TI - Prognostic prediction and treatment strategy in hepatocellular carcinoma. PMID- 11870364 TI - Independent and overlapping transcriptional activation during liver development and regeneration in mice. AB - Liver development and regeneration share the requirement for simultaneous proliferation and acquisition of highly specialized cellular functions. However, little is known about molecules with regulatory roles in both processes. We hypothesized that transcriptional reprogramming induced by regeneration recapitulates that of developing liver. To address this hypothesis, we determined global hepatic gene expression at embryonic day 14.5, postnatal day 14, and 6 to 24 hours following partial hepatectomy using microarrays containing 8,635 cDNAs. Analysis of genes overexpressed during these conditions revealed 3 unique expression patterns. The first was predominantly signature gene clusters specific for each growth phase. Major groups were hematopoiesis-related genes in embryonic livers, metabolic genes during postnatal liver development, and growth/inflammation and metabolic genes during regeneration. The second pattern consisted of dual overexpression during regeneration and at least one phase of development. Consistent with potential regulatory roles in liver growth, most of these transcripts control cell-cell contact, membrane trafficking, cell growth, metabolism, and inflammatory response. The third pattern, revealed by surveying their expression across 76 hepatic and extra-hepatic tissues, uncovered a restricted temporospatial pattern of liver overexpression for CD14, orosomucoid 1, hepcidin, Spi 2.1, Ith3, and Tim-44. In conclusion, these results provide a basis for the identification of gene and gene groups that play critical roles at different phases of liver development and regeneration, and underscore the importance of maintaining metabolic demands during organ growth. PMID- 11870365 TI - A20 protects mice from D-galactosamine/lipopolysaccharide acute toxic lethal hepatitis. AB - Apoptosis of hepatocytes is a seminal feature of fulminant hepatic failure. We show that the anti-apoptotic protein A20 is upregulated in hepatocytes by pro inflammatory stimuli and functions to protect from apoptosis and limit inflammation by inhibiting NF-kappaB. Adenoviral mediated hepatic expression of A20 in BALB/c mice yields an 85% survival rate in the D-galactosamine (D gal)/lipolysaccharide (LPS) model of acute toxic hepatitis compared with 15% to 20 % in control mice. Expression of A20 preserves normal liver function as assessed by prothrombin time. The protective effect of A20 is independent of tumor necrosis factor (TNF) inhibition. Maintaining high circulating TNF levels may be advantageous for liver regeneration. Our data supports this hypothesis as evidenced by increased proliferating cell nuclear antigen (PCNA) expression in the livers of mice expressing A20 compared with a dominant negative mutant of the TNF receptor (TNF-R), 6 hours following D-gal/LPS administration. In conclusion, these results qualify A20 as part of a physiologic, protective response of hepatocytes to injury and a promising gene therapy candidate for clinical applications aimed at preventing and treating viral and toxic fulminant hepatic failure. PMID- 11870366 TI - Role of mitochondrial permeability transition in diclofenac-induced hepatocyte injury in rats. AB - Hepatotoxicity of diclofenac has been known in experimental animals and humans but its mechanism has not been fully understood. The present study examined the role of mitochondrial permeability transition (MPT) in the pathogenesis of diclofenac-induced hepatocyte injury by using isolated mitochondria and primary culture hepatocytes from rats. Incubation of energized mitochondria with succinate in the presence of Ca(2+) and diclofenac resulted in mitochondrial swelling, leakage of accumulated Ca(2+), membrane depolarization, and oxidation of nicotinamide adenine dinucleotide phosphate and protein thiol. All of these phenomena were suppressed by coincubation of the mitochondria with cyclosporin A, a typical inhibitor of MPT, showing that diclofenac opened the MPT pore. It was also suggested that reactive oxygen species probably generated during mitochondrial respiration and/or voltage-dependent mechanism was involved in MPT, which are proposed as mechanisms of MPT by uncouplers of mitochondrial oxidative phosphorylation. Culture of hepatocytes for 24 hours with diclofenac caused a decrease in cellular ATP, leakage of lactate dehydrogenase and membrane depolarization. The hepatocyte toxicity thus observed was attenuated by coincubation of the hepatocytes with cyclosporin A and verapamil, a Ca(2+) channel blocker. In conclusion, these results showed the important role of MPT in pathogenesis of hepatocyte injury induced by diclofenac and its possible contribution to human idiosyncratic hepatotoxicity. PMID- 11870367 TI - COX-2 inhibits Fas-mediated apoptosis in cholangiocarcinoma cells. AB - Fas expression has been shown to negatively regulate the progression of cholangiocarcinoma cells in xenografts. However, many human cholangiocarcinomas express Fas, suggesting these cancers have developed mechanisms to inhibit Fas mediated apoptosis. Cyclooxygenase-2 (COX-2), which generates prostanoids, is expressed by many cholangiocarcinomas. Therefore, our aim was to determine whether COX-2 expression inhibits death receptor--mediated apoptosis in KMBC cells, a cholangiocarcinoma cell line. These cells express messenger RNA for the death receptors Fas, tumor necrosis factor receptor 1 (TNF-R1), death receptor 4 (DR4), and DR5. Agonists for these death receptors, CH-11, TNF-alpha, and TRAIL all induced apoptosis. However, COX-2, whether induced by proinflammatory cytokines or transient transfection, only significantly inhibited Fas-mediated apoptosis. The COX-2 inhibitor NS-398 restored Fas-mediated apoptosis in COX-2 transfected cells. Prostaglandin E2 reduced apoptosis and mitochondrial depolarization after treatment with the Fas agonist CH-11. Of a variety of antiapoptotic proteins examined, COX-2/prostaglandin E2 only increased expression of Mcl-1, an antiapoptotic member of the Bcl-2 family. In conclusion, these data suggest that prostanoid generation by COX-2 specifically inhibits Fas-mediated apoptosis, likely by up-regulating Mcl-1 expression. Pharmacologic inhibition of COX-2 may be useful in augmenting Fas-mediated apoptosis of cholangiocarcinoma cells. PMID- 11870368 TI - Cold-induced apoptosis of rat liver cells in University of Wisconsin solution: the central role of chelatable iron. AB - Although University of Wisconsin (UW) solution aims at the prevention of cold induced cell injury, it failed to protect against cold-induced apoptosis of hepatocytes and liver endothelial cells: when incubated in UW solution at 4 degrees C for 24 hours and subsequently rewarmed at 37 degrees C, 72% +/- 8% of rat hepatocytes and 81% +/- 5% of liver endothelial cells lost viability. In both cell types, the observed cell damage occurred under an apoptotic morphology; it appeared to be mediated by a rapid increase in the cellular chelatable iron pool by a factor > or =2 (as determined in hepatocytes) and subsequent formation of reactive oxygen species (ROS). Consequently, this cell injury was decreased by iron chelators to 6 to 25% (hepatocytes) and 4% +/- 2% (liver endothelial cells). Deferoxamine nearly completely inhibited the occurrence of apoptotic morphology in both cell types. In liver endothelial cells, cold-induced apoptosis occurring during rewarming after 24 hours of cold incubation in UW solution was far more pronounced than in cell culture medium (loss of viability: 81% +/- 5% vs. 28% +/- 13%), but viability could even be maintained for 2 weeks of cold incubation by use of deferoxamine. In conclusion, this pathological mechanism might be an explanation for the strong endothelial cell injury known to occur after cold preservation. With regard to the extent of this iron-mediated injury, addition of a suitable iron chelator to UW solution might markedly improve the outcome of liver preservation. PMID- 11870369 TI - Differentiation-dependent and subset-specific recruitment of T-helper cells into murine liver. AB - It has been suggested that the liver traps and deletes activated and potentially harmful T cells, especially of the CD8(+) subset, providing mechanisms to limit systemic immune responses. It is unknown whether this also applies to CD4(+) T cells. In this study, we show that activated stages of CD4(+) T cells were trapped in the liver on intraportal injection. Intravital microscopy showed an immediate adhesion of activated CD4(+) T cells within periportal sinusoids after intraportal injection. Furthermore, we detected high frequencies of interferon gamma (IFN-gamma)-- (Th1) and interleukin 4 (IL-4)-- (Th2) synthesizing effector cells in the liver. Transfer experiments were performed to identify those phenotypes showing specific retention in the liver. Our data show that effector stages and activated cells in general are more efficiently recruited into the liver than resting CD4(+) T cells, similar to what has previously been shown for CD45RB(low) memory cells. In addition, we observed a certain preference for Th1 polarized cells to be trapped by the liver. However, the actual cytokine producing cells did not specifically enrich among the total population. In conclusion, these data indicate that the liver acts as a filter for activated and memory/effector cells. Cells trapped in the liver might subsequently undergo modulatory influences exerted by the postulated specific microenvironment of the liver. PMID- 11870370 TI - The cyclooxygenase system participates in functional mdr1b overexpression in primary rat hepatocyte cultures. AB - Overexpression of mdr1-type P-glycoproteins (P-gps) is thought to contribute to primary chemotherapy resistance of untreated hepatocellular carcinoma. However, mechanisms of endogenous multidrug resistance 1 (mdr1) gene activation still remain unclear. Because recent studies have demonstrated overexpression of cyclooxygenase-2 (COX-2) in hepatocytes during early stages of hepatocarcinogenesis, we investigated whether the COX system, which catalyzes the rate-limiting step in prostaglandin synthesis, participates in mdr1 gene regulation. In the present study, primary rat hepatocyte cultures, exhibiting time-dependent mdr1b overexpression, demonstrated basal COX-2 and COX-1 mRNA expression and liberation of prostaglandin E(2) (PGE(2)), indicative of an active COX-dependent arachidonic acid metabolism. PGE(2) accumulation in culture supernatants was further enhanced by arachidonic acid (1mumol/L) and epidermal growth factor (EGF) (16 nmol/L). PGE(2) and prostaglandin F(2alpha) (PGF(2)alpha) (3-6mug/mL), added directly to the culture medium, significantly up-regulated intrinsic mdr1b mRNA overexpression and mdr1-dependent transport activity. Up regulation was maximal after 3 days of culture. Like prostaglandins, the COX substrate, arachidonic acid, also induced mdr1b gene expression. Apart from this, structurally different COX inhibitors (indomethacin, meloxicam, NS-398) mediated significant inhibition of time-dependent and EGF-induced mdr1b mRNA overexpression, resulting in enhanced intracellular accumulation of the mdr1 substrate, rhodamine 123 (Rho123). Thus, the present data support the conclusion that the release of prostaglandins through activation of the COX system participates in endogenous mdr1b gene regulation. COX-2 inhibition might constitute a new strategy to counteract primary mdr1-dependent chemotherapy resistance. PMID- 11870371 TI - Farnesoid X receptor and bile salts are involved in transcriptional regulation of the gene encoding the human bile salt export pump. AB - The bile salt export pump (BSEP or ABCB11) mediates the adenosine triphosphate dependent transport of bile salts across the canalicular membrane of the hepatocyte. Mutations in the corresponding ABCB11 gene cause progressive familial intrahepatic cholestasis type 2. The aim of this study was to investigate the regulation of human ABCB11 gene transcription by bile salts. First, a 1.7 kilobase human ABCB11 promoter region was cloned. Sequence analysis for possible regulatory elements showed a farnesoid X receptor responsive element (FXRE) at position minus sign180. The farnesoid X receptor (FXR) functions as a heterodimer with the retinoid X receptor alpha (RXRalpha) and can be activated by the bile salt chenodeoxycholic acid (CDCA). Luciferase reporter gene assays showed that the ABCB11 promoter is positively controlled by FXR, RXRalpha, and bile salts in a concentration-dependent manner. Mutation of the FXRE strongly represses the FXR dependent induction. Second, endogenous ABCB11 transcription regulation was studied in HepG2 cells, stably expressing the rat sodium-dependent taurocholate transporter (rNtcp) cells. ABCB11 expression was induced by adding bile salts to the culture medium, and this effect was maximized by combining it with cotransfection of rFxr and hRXRalpha. Reducing endogenous FXR levels using RNA interference fully repressed the bile salt-induced ABCB11 expression. In conclusion, these results show that FXR is required for the bile salt-dependent transcriptional control of the human ABCB11 gene and that the cellular amount of FXR is critical for the level of activation of ABCB11 transcription. PMID- 11870372 TI - Dissolution of cholesterol gallstones in mice by the oral administration of a fatty acid bile acid conjugate. AB - Gallstones, mostly cholesterol stones, affect some 15% of the population. Oral bile salts dissolve human cholesterol gallstones, but with low efficacy, and surgery remains the main therapeutic option. Fatty acid bile acid conjugates (FABACs) were shown to prevent formation of cholesterol gallstones in experimental animals. The aim of this study was to test whether these compounds could dissolve preexisting cholesterol gallstones via oral administration. Inbred, gallstone-susceptible C57J/L mice were given a lithogenic diet for 2 months, and the presence of gallstones was ascertained. The mice were then switched to a regular diet while part of them were given in addition C20-FABAC, by gavage, at a dose of 0.5 or 3 mg per animal per day. All mice tested had cholesterol gallstones after 2 months on the lithogenic diet. In study I, after 2 months on the regular diet, 3 of 4 (75%) of the controls had gallstones, whereas none of the 6 FABAC-fed animals (3 mg/d) had stones (P =.033). In study II, evaluating 2 FABAC doses, after 2 months on the regular diet, 8 of 8 (100%) of the controls had gallstones, which were found in 2 of 7 (28%) and 1 of 8 (12%) of the mice supplemented with 0.5 mg/d (P =.007) or 3 mg/d (P =.001) FABAC, respectively. On a molar basis, the dose of 0.5 mg FABAC is equivalent to 14 mg/kg/d of a bile acid. In conclusion, FABACs given orally can dissolve preexisting cholesterol gallstones in mice. This was accomplished with a dose of FABAC equivalent to the dose of bile acids used in human gallstone dissolution. PMID- 11870373 TI - Change in portal flow after liver transplantation: effect on hepatic arterial resistance indices and role of spleen size. AB - Information on changes in splanchnic hemodynamics after liver transplantation is incomplete. In particular, data on long-term changes are lacking, and the relationship between changes in arterial and portal parameters is still under debate. The effect of liver transplantation on splanchnic hemodynamics was analyzed with echo-Doppler in 41 patients with cirrhosis who were followed for up to 4 years. Doppler parameters were also evaluated in 7 patients transplanted for acute liver failure and in 35 controls. In cirrhotics, portal blood velocity and flow increased immediately after transplantation (from 9.1 plus minus 3.7 cm/sec to 38.3 plus minus 14.6 and from 808 plus minus 479 mL/min to 2,817 plus minus 1,153, respectively, P <.001). Hepatic arterial resistance index (pulsatility index) also augmented (from 1.36 plus minus 0.32 to 2.34 plus minus 1.29, P <.001) and was correlated with portal blood velocity and flow. The early changes in these parameters were related, in agreement with the hepatic buffer response theory. Portal flow returned to normal values after 2 years. Superior mesenteric artery flow normalized after 3 to 6 months. Splenomegaly persisted after 4 years, when spleen size was related to portal blood flow. In 7 patients transplanted for acute liver failure, portal flow, and hepatic arterial resistance index were normal after transplantation. In conclusion, a high portal flow was present in cirrhotics until 2 years after transplantation, probably because of maintenance of elevated splenic flow. An early increase in hepatic arterial resistance indices is a common finding, but it is transient and is related to the increase in portal blood flow. A normal time course of portal-hepatic hemodynamics was detected in patients transplanted for acute liver failure. PMID- 11870374 TI - Endoscopic treatment versus endoscopic plus pharmacologic treatment for acute variceal bleeding: a meta-analysis. AB - Endoscopic therapy, involving either injection sclerosis or band ligation, is considered the intervention of first choice for acute variceal bleeding (AVB). Pharmacologic agents have also been shown to be highly effective in the control of the bleeding episode. The purpose of this meta-analysis was to assess whether vasoactive drugs may improve the efficacy of endoscopic therapy (injection sclerosis or band ligation) in the control of AVB and thus increase survival rates. Computer databases and scientific meeting abstracts from 1994 to 2001 were used to search for randomized trials that compared the combined use of endoscopic and drug therapy with endoscopic therapy alone in the control of AVB. Eight trials involving 939 patients fulfilled the selection criteria and the following evaluated by standard meta-analysis methods: initial hemostasis, 5-day hemostasis, 5-day mortality, and adverse events. Combined treatment improved initial control of bleeding (relative risk [RR], 1.12; 95% confidence interval (CI), 1.02-1.23), and 5-day hemostasis (RR, 1.28; 95% CI, 1.18-1.39), with numbers of patients needed to treat (NNT) of 8 and 5, respectively. The difference in favor of combined treatment remained significant when trials that used drugs other than octreotide or that included a low proportion of alcoholic patients (<40%) or high-risk cirrhotic patients (<35%) were excluded. Mortality was not significantly decreased by combined therapy (RR, 0.73; 95% CI, 0.45 1.18). Severe adverse events were similar in both groups. In conclusion, in patients with AVB, pharmacologic agents improve the efficacy of endoscopic therapy to achieve initial control of bleeding and 5-day hemostasis, yet fail to affect mortality. PMID- 11870375 TI - Recombinant factor VIIa improves clot formation but not fibrolytic potential in patients with cirrhosis and during liver transplantation. AB - Cirrhosis is associated with a bleeding tendency, which is particularly pronounced during orthotopic liver transplantation (OLT). A novel approach to treating the bleeding diathesis of patients with cirrhosis is administration of recombinant factor VIIa (rFVIIa). This study examined whether the efficacy of rFVIIa in cirrhosis might be explained in part by enhanced down-regulation of fibrinolysis by thrombin-activatable fibrinolysis inhibitor (TAFI). Addition of therapeutic or supratherapeutic doses of rFVIIa to plasma of 12 patients with stable cirrhosis did not result in a prolongation of clot lysis time, though clotting times were significantly reduced. Also, clot lysis assays of plasma samples taken during and after OLT, which was performed with or without a single bolus dose of rFVIIa, did not show any effect of rFVIIa on plasma fibrinolytic potential. In conclusion, this study shows no evidence for an antifibrinolytic effect of rFVIIa in cirrhotic patients or in patients undergoing OLT. PMID- 11870376 TI - Intestinal mucosal alterations in experimental cirrhosis in the rat: role of oxygen free radicals. AB - Cirrhosis is associated with altered gastrointestinal function, and bacterial translocation from the gut plays an important role in the etiology of spontaneous bacterial peritonitis (SBP) seen in this condition. Although alterations in gut motility and intestinal permeability are recognized in cirrhosis, the intestinal damage at the cellular and subcellular levels is not well understood. This study looked at the mucosal alterations in experimental cirrhosis and the role of oxygen free radicals in this process. It was shown that cirrhosis results in oxidative stress in the intestine, as seen by increased xanthine oxidase (XO) activity and altered antioxidant status. Cirrhosis also affects enterocyte mitochondrial function, as assessed by respiratory control ratio, swelling, and calcium flux. Increased lipid peroxidation of the brush border membranes (BBMs) was seen along with altered intestinal transport. In conclusion, this study shows that intestinal mucosal alterations are seen in experimental cirrhosis and are possibly mediated by oxidative stress. PMID- 11870377 TI - Hepatic arterial buffer response in patients with advanced cirrhosis. AB - Hepatic arterial buffer response (HABR) is considered an important compensatory mechanism to maintain perfusion of the liver by hepatic arterial vasodilation on reduction of portal venous perfusion. HABR has been suggested to be impaired in patients with advanced cirrhosis. In patients with hepatopetal portal flow, placement of a transjugular intrahepatic portosystemic shunt (TIPS) reduces portal venous liver perfusion. Accordingly, patients with severe cirrhosis should have impaired HABR after TIPS implantation. Therefore, the aim of this study was to investigate the effect of TIPS on HABR as reflected by changes in resistance index (RI) of the hepatic artery. A total of 366 patients with cirrhosis (Child Pugh class A, 106; class B, 168; class C, 92) underwent duplex Doppler ultrasonographic examination with determination of RI and maximal flow velocity in the portal vein before and 1 month after TIPS placement. Portosystemic pressure gradient was determined before and after TIPS placement. In 29 patients with hepatofugal portal blood flow, RI was significantly lower than in 337 patients with hepatopetal flow (0.63 plus minus 0.02 vs. 0.69 plus minus 0.01; P <.001). TIPS induced a significant decrease of the RI in patients with hepatopetal flow (RI, 0.69 plus minus 0.01 before vs. 0.64 plus minus 0.01 after TIPS; P =.001) but not in patients with hepatofugal flow (RI, 0.63 plus minus 0.02 before vs. 0.63 plus minus 0.02 after TIPS; NS). This response was not dependent on the Child-Pugh class. In conclusion, our results suggest that some degree of HABR is preserved even in patients with advanced cirrhosis with significant portal hypertension. PMID- 11870378 TI - Risk factors of fibrosis in alcohol-induced liver disease. AB - In patients with nonalcoholic steatohepatitis (NASH), age, obesity, and diabetes mellitus are independent predictors of the degree of fibrosis. The relative risk for fibrosis adjusted for sex was also associated with increasing grade of Perls stain. The aim of this study was to determine whether the risk factors for fibrosis described in NASH are also risk factors in alcohol-induced liver disease. A total of 268 alcoholic patients with negative hepatitis B virus and hepatitis C virus serology underwent liver biopsy. Fibrosis was assessed semiquantitatively by a score fluctuating between 0 to 8. Liver iron overload was assessed by Perls staining and graded in 4 classes. We have used multivariate regression with partial correlation analysis to assess the variability of fibrosis score according to the value of 7 variables: sex, age, body mass index (BMI) in the past year before the hospitalization when the patient was asymptomatic, daily alcohol intake over the past 5 years, total duration of alcohol abuse, Perls grade, and blood glucose level. In the multivariate regression, fibrosis score was positively correlated with age (P =.001), BMI (P =.002), female sex (P <.05), Perls grade (P <.05), and blood glucose level (P <.05). Twenty percent of the variability of fibrosis score was explained by the 7 variables. In conclusion, after adjustment for daily alcohol intake and total duration of alcohol abuse, BMI, Perls grade, and blood glucose are also independent risk factors for fibrosis in alcohol-induced liver disease, raising therapeutic implications for the management of these patients. PMID- 11870379 TI - Histologic and biochemical changes during the evolution of chronic rejection of liver allografts. AB - Criteria for histologic diagnosis of chronic rejection (CR) are based on changes seen late in the disease process that are likely to be irreversible and unresponsive to treatment. Changes occurring during the evolution of CR are less clearly defined. The serial biopsy specimens, failed allografts, and biochemical profiles of 28 patients who underwent retransplantation for CR were examined with the aim of identifying histologic and biochemical features that were present during the early stages of CR. For each case, a point of acute deterioration in liver function tests (LFTs) was identified ("start time" [ST]) that subsequently progressed to graft failure. Biopsy specimens before, at the time of ("start biopsy" [SB]), and after the ST were assessed histologically, and findings were correlated with the biochemical changes. CR resulted from acute rejection (AR) that did not resolve. Centrilobular necroinflammation (CLNI) associated with an elevated aspartate transaminase (AST) level and portal tract features of AR were present at the start. Portal AR features resolved, CLNI persisted, AST level remained elevated, and bilirubin and alkaline phosphatase levels progressively increased throughout the evolution of CR. Portal tracts also showed a loss of small arterial and bile duct branches, with arterial loss occurring early and bile duct loss as a later progressive lesion. Foam cell arteriopathy was rarely seen in needle biopsy specimens. In conclusion, findings from this study may help identify patients at risk of progressing to graft loss from CR at a stage when the disease process is potentially reversible and amenable to treatment. PMID- 11870380 TI - Time-dependent Cox regression model is superior in prediction of prognosis in primary sclerosing cholangitis. AB - More precise prognostic models are needed for prediction of survival in patients with primary sclerosing cholangitis (PSC), particularly for the selection of candidates for liver transplantation. The aim of this study was to develop a time dependent prognostic model for the calculation of updated short-term survival probability in PSC. Consecutive clinical and laboratory follow-up data from the time of diagnosis were collected from the files of 330 PSC patients from 5 European centers, followed for a median of 8.4 years since diagnosis. Time-fixed and time-dependent Cox regression analyses, as well as the additive regression model, were applied. The reliability of the models was tested by a cross validation procedure. Bilirubin (on a logarithmic scale), albumin, and age at diagnosis of PSC were identified as independent prognostic factors in multivariate analysis of both the time-fixed and the time-dependent Cox regression models. The importance of bilirubin was more pronounced in the time dependent model (hazard ratio [HR], 2.84) than in the time-fixed analysis (hazard ratio, 1.51). The additive regression model indicated that once the patients survive beyond the first 5 years, the impact on prognosis of albumin at diagnosis ceases. The time-dependent prognostic model was superior to the time-fixed variant in assigning low 1-year survival probabilities to patients that actually survived less than 1 year. In conclusion, a time-dependent Cox regression model has the potential to estimate a more precise short-term prognosis in PSC compared with the traditional time-fixed models. PMID- 11870381 TI - Antibodies to conformational epitopes of soluble liver antigen define a severe form of autoimmune liver disease. AB - Prevalence and clinical relevance of antibodies to soluble liver antigen (tRNP((Ser)Sec)/SLA) in autoimmune hepatitis (AIH) have been investigated using partially purified or prokaryotically expressed antigen. The aim of this study was to improve the detection of anti-tRNP((Ser)Sec)/SLA by establishing an immunoassay that was able to identify antibodies directed to conformational epitopes and to investigate the clinical implication of this autoantibody in autoimmune liver disease. By using eukaryotically expressed tRNP((Ser)Sec)/SLA as target in a radioligand assay (RLA), 81 patients with autoimmune liver disease (AILD) (33 type 1 AIH, 31 type 2 AIH, and 17 autoimmune sclerosing cholantitis [ASC]), 147 pathologic, and 56 healthy controls were investigated. RLA results were compared with those obtained using a commercial enzyme-linked immunosorbent assay (ELISA) and immunoblot. Reactivity to tRNP((Ser)Sec)/SLA was present in 58% of patients with type 1 and type 2 AIH, 41% with ASC, but in only 3 pathologic controls. RLA was similarly disease-specific but remarkably more sensitive than ELISA and immunoblot. A prospective study showed that anti-tRNP((Ser)Sec)/SLA positive patients run a severe clinical course, having worse histology, needing longer to achieve remission, relapsing and requiring liver transplantation or dying more frequently than anti-tRNP((Ser)Sec)/SLA negative patients. Anti tRNP((Ser)Sec)/SLA production was favored by the possession of DR3 and A1-B8-DR3 in AIH type 1 and ASC, and prevented by the possession of A2 in all 3 types of AILD, particularly in type 2 AIH. In conclusion, anticonformational tRNP((Ser)Sec)/SLA reactivity is frequent in type 1 and type 2 AIH and ASC, defining patients with a worse prognosis. PMID- 11870382 TI - Varied assembly and RNA editing efficiencies between genotypes I and II hepatitis D virus and their implications. AB - The mechanisms that link genotypes of hepatitis D virus (HDV) with clinical outcomes have not yet been elucidated. Genotypic variations are unevenly distributed along the sequences of hepatitis delta antigens (HDAgs). Of these variations, the packaging signal at the C-terminus has a divergence of 74% between genotypes I and II. In this report, we address the issue of whether these high variations between genotypes affect assembly efficiency of HDV particles and editing efficiency of RNA. Viral package systems of transfection with expression plasmids of hepatitis B surface antigen and HDAgs or whole genomes of HDV consistently indicate that the package efficiency of genotype I HDV is higher than that of genotype II. Segment swapping of large-form HDAg indicates that the C-terminal 19-residue region plays a key role for the varied assembly efficiencies. Also, the editing efficiency of genotype I HDV is higher than that of genotype II. The nucleotide and structural changes surrounding the editing site may explain why genotype II HDV has a low RNA editing efficiency. The findings of in vitro assembly systems were further supported by the observations that patients infected with genotype II had significantly lower alanine transaminase (ALT) levels, more favorable outcomes (P <.05), and a trend to have lower serum HDV RNA levels as compared with those infected with genotype I HDV (P =.094). In conclusion, genotype II HDV secretes fewer viral particles than genotype I HDV does, which in turn may reduce the extent of infection of hepatocytes and result in less severe hepatic inflammation. PMID- 11870383 TI - Clinical evaluation (phase I) of a combination of two human monoclonal antibodies to HBV: safety and antiviral properties. AB - Treatment of chronic hepatitis B virus (HBV) infection with interferon alfa and lamivudine is characterized by lack of viral clearance, loss of response, or emergence of drug-resistant mutants. Thus, new and multiple drug approaches are needed. We have developed two fully human monoclonal antibodies, directed against different epitopes of hepatitis B surface antigen (HBsAg) that bind to all major HBV subtypes. A phase I clinical study was conducted to evaluate the safety, tolerability, and efficacy of a mixture of these two monoclonal antibodies, HBV AB(XTL). A total of 27 chronic HBV patients were enrolled. In part A of the study 15 patients in 5 cohorts received a single intravenous infusion of antibodies with doses ranging from 0.26 mg (260 IU) to 40 mg (40,000 IU). All patients completed 16 weeks of follow-up. In the second part of the study (part B), 12 patients in 4 cohorts received 4 weekly infusions of 10, 20, 40, or 80 mg each of HBV-AB(XTL) and were followed for 4 additional weeks. Administration of antibodies was well tolerated. Patients administered doses at an Ab:Ag molar ratio of 1:2 to 1:20 showed a rapid and significant decrease in HBsAg to undetectable levels, with a corresponding reduction of HBV-DNA levels. In part B, HBV-AB(XTL) induced a significant reduction in both HBsAg and HBV-DNA levels repeatedly after administration. In conclusion, these data suggest that HBV AB(XTL) binds HBV particles and reduces serum viral titers and HBsAg levels. HBV AB(XTL) could be combined with other monotherapies that are currently used to treat HBV carriers. PMID- 11870384 TI - Hepatitis C virus kinetics during and immediately after liver transplantation. AB - The study of hepatitis C virus (HCV) kinetics after liver transplantation (LT) might be important to design strategies to prevent HCV infection of the graft. We analyzed HCV kinetics during and immediately after LT in 20 consecutive patients undergoing LT for HCV-related cirrhosis. HCV RNA was quantified in blood samples obtained at regular intervals before, during, and after transplantation. HCV-RNA concentrations decreased in 18 of 20 patients during the anhepatic phase (mean decay slope -0.92, mean HCV elimination half-life 2.2 hours). We found a significant correlation between the HCV viral load decay and the blood loss during the anhepatic phase, indicating that the observed HCV clearance rates are maximum estimates. In fact, in 1 patient with an unusually long anhepatic phase of 20 hours and with minimum blood loss, the HCV elimination half-life was 10.3 hours. Eight to 24 hours after graft reperfusion a sharp decrease in HCV viral load occurred in 19 patients (mean decay slope -0.34, mean HCV elimination half life 3.44 hours). HCV RNA became undetectable in only 1 patient. During the following days, HCV-RNA concentrations increased rapidly in 10 patients (mean HCV doubling time 13.8 hours), remained at similar levels in 4, and continued to decrease in 6. The only variable associated with a second-phase viral load decay was the absence of corticosteroids as part of the immunosuppressive regimen. In conclusion, a sharp decrease in HCV viral load occurs during the anhepatic phase and immediately after graft reperfusion, most likely owing to a lack of virion production and hepatic viral clearance. HCV infection of the graft, however, is an extremely dynamic process and viral replication begins a few hours after LT. PMID- 11870385 TI - Hepatic HCV RNA before and after treatment with interferon alone or combined with ribavirin. AB - The clinical use of measuring hepatic hepatitis C virus (HCV) RNA before and after therapy in patients with chronic hepatitis C has been assessed in a number of small clinical trials. Viral clearance from the liver may be a better marker of long-term response than eradication of serum HCV RNA. The aim of this study was to evaluate quantitative hepatic HCV-RNA measurements before and after antiviral therapy. Two thousand eighty-nine chronic hepatitis C patients were enrolled in 3 published clinical trials evaluating interferon alfa-2b alone or with ribavirin either as initial therapy or for interferon relapse. Hepatic HCV RNA quantitation was performed with a modified reverse-transcription polymerase chain reaction (RT-PCR) before and 24 weeks after therapy in 951 and 1,316 patients, respectively. Pretherapy hepatic HCV-RNA concentrations correlated best with serum HCV-RNA concentrations (R =.236, P =.0001) and negatively correlated with alanine transaminase (ALT) values (-0.178, P =.0001), duration of infection (-0.09, P =.02), parenchymal injury (-0.135, P =.0001), histologic activity index (HAI) inflammatory score (-0.085, P =.01), Knodell fibrosis score (-0.072, P =.03), and body weight (-0.078, P =.02). In paired liver biopsy specimens (n = 534), change in hepatic HCV RNA correlated with the change in the HAI (R =.346, P =.0001). Of 400 sustained virologic responders (SVR), 393 (98%) had undetectable hepatic HCV RNA, whereas 7 (2%) had detectable hepatic HCV RNA; 5 have been followed and 2 have had reappearance of serum HCV RNA 12 months after therapy. In conclusion, measurement of hepatic HCV RNA before or after therapy reflects changes observed in serum HCV RNA, and correlates inversely with hepatic inflammation and fibrosis, but otherwise has minimal clinical use. PMID- 11870386 TI - Interferon-gamma inhibits replication of subgenomic and genomic hepatitis C virus RNAs. AB - Persistent infection with hepatitis C virus (HCV) is a major cause of chronic hepatitis, liver cirrhosis, and hepatocellular carcinoma. All treatments known so far rely on the antiviral activity of interferon alfa (IFN-alpha) that is given alone or in combination with ribavirin. Unfortunately, only a fraction of the patients clear the virus during therapy and for those who do not respond there is currently no alternative treatment. Selectable subgenomic HCV RNAs (replicons) have been recently used to investigate the effect of IFN-alpha on HCV replication. However, it has not yet been analyzed whether other cytokines also play a role in the innate immune response against HCV. Here we show that IFN gamma inhibits protein synthesis and RNA replication of subgenomic and genomic HCV replicons. We further show that the inhibitory action of IFN-gamma does not rely on the production of nitric oxide or the depletion of tryptophan. In conclusion, our results suggest that cytotoxic T cells and natural killer cells may contribute to HCV clearance not only by cell killing but also by producing IFN-gamma, thereby enhancing the intracellular inhibition of viral replication. PMID- 11870387 TI - Relationship of health-related quality of life to treatment adherence and sustained response in chronic hepatitis C patients. AB - Interferon therapy may exacerbate health-related quality of life (HRQL) deficits associated with hepatitis C virus (HCV) early in the course of therapy. Treatment with polyethylene glycol-modified interferon (peginterferon) alfa-2a (40 kd) provides improved sustained response over interferon alfa-2a, but its effect on HRQL is unknown. The objective of this study was to (1) evaluate the effect of sustained virologic response on HRQL in patients with HCV and (2) determine whether impairment of HRQL during treatment contributes to early treatment discontinuation. Data consisted of a pooled secondary analysis of patients (n = 1,441) across 3 international, multicenter, open-label, randomized studies that compared peginterferon alfa-2a (40 kd) with interferon alfa-2a. ANCOVA was used to examine the effect of sustained virologic response on HRQL. Repeated-measures mixed-models ANCOVA was used to compare Fatigue Severity Scale (FSS) and SF-36 scores during treatment by treatment group. Logistic regression analysis was used to examine the association between changes at baseline in on-treatment HRQL and early treatment discontinuation. Sustained virologic response was associated with marked improvements from baseline to end of follow-up in all subjects, including patients with cirrhosis. During treatment, patients receiving peginterferon alfa 2a (40 kd) had statistically significantly better scores on both the SF-36 and FSS. Baseline to 24-week changes in fatigue and SF-36 mental and physical summary scores significantly predicted treatment discontinuation. In conclusion, sustained virologic response is associated with improvements in quality of life in patients with or without advanced liver disease. This parameter may be an important consideration in maximizing treatment adherence. PMID- 11870388 TI - Modeling the hepatitis C virus epidemic in France using the temporal pattern of hepatocellular carcinoma deaths. AB - Deuffic et al. developed a compartmentalized model that characterized the evolution and spread of the hepatitis C virus (HCV) within France. There were various parameters defining the age- and sex-dependent transition probabilities between chronic hepatitis and cirrhosis in need of determination to completely specify their model. These were estimated by means of a weighted least-squares procedure that was executed numerically. The objective function used was based on the distribution of the age at death from hepatocellular carcinoma (HCC) rather than the temporal pattern of deaths due to HCC from 1979 to 1995. In this report, we investigate the impact of using an objective function based on the temporal pattern of deaths. We show that the dynamics of the epidemic can be quite different, in particular, short-term prediction of HCC deaths by HCV infection and times to death from onset of disease. PMID- 11870389 TI - Hepatic encephalopathy--definition, nomenclature, diagnosis, and quantification: final report of the working party at the 11th World Congresses of Gastroenterology, Vienna, 1998. AB - Research on hepatic encephalopathy is hampered by the imprecise definition of this disabling complication of liver disease. Under this light, the Organisation Mondiale de Gastroenterologie commissioned a Working Party to reach a consensus in this area and to present it at the 11th World Congress of Gastroenterology in Vienna (1998). The Working Party continued its work thereafter and now present their final report. In summary, the Working Party has suggested a modification of current nomenclature for clinical diagnosis of hepatic encephalopathy; proposed guidelines for the performance of future clinical trials in hepatic encephalopathy; and felt the need for a large study to redefine neuropsychiatric abnormalities in liver disease, which would allow the diagnosis of minimal (subclinical) encephalopathy to be made on firm statistical grounds. In the interim, it proposes the use of a psychometric hepatic encephalopathy score, based on the result of 5 neuropsychologic tests. Finally, the need for a careful evaluation of the newer neuroimaging modalities for the diagnosis of hepatic encephalopathy was stressed. PMID- 11870390 TI - Of mice and men: a small animal model of hepatitis C virus replication. PMID- 11870391 TI - A new twist to translation initiation of the hepatitis C virus. PMID- 11870392 TI - Microarray analysis of hepatic-regulated gene expression: specific applications and nonspecific problems. PMID- 11870393 TI - Risk of HAART therapy in hepatitis C. PMID- 11870394 TI - Liver regeneration in obese mice with fatty livers: does the impairment have relevance for other types of fatty liver disease? PMID- 11870396 TI - Organic anion transporting polypeptides, cholestasis, and nuclear receptors. PMID- 11870398 TI - Along came a spider. PMID- 11870409 TI - Calcium and vitamin D in osteoporosis: chairman's introduction to proceedings. PMID- 11870410 TI - The importance of calcium intake for lifelong skeletal health. PMID- 11870411 TI - Calcium and vitamin D in osteoporosis: supplementation or treatment? PMID- 11870412 TI - Influence of daily regimen calcium and vitamin D supplementation on parathyroid hormone secretion. AB - Calcium and vitamin D supplementation has been shown to reduce secondary hyperparathyroidism and play a role in the management of senile osteoporosis. In order to define the optimal regimen of calcium and vitamin D supplementation to produce the maximal inhibition of parathyroid hormone secretion, we have compared the administration of a similar amount of Ca and vitamin D, either as a single morning dose or split in two doses, taken 6 hours apart. Twelve healthy volunteers were assigned to three investigational procedures, at weekly intervals. After a blank control procedure, when they were not exposed to any drug intake, they received two calcium-vitamin D supplement regimens including either two doses of Orocal D3 (500 mg Ca and 400 IU vitamin D) 6 hours apart or one water-soluble effervescent powder pack of Cacit D3 in a single morning dose (1000 mg Ca and 880 IU vitamin D). During the three procedures (control and the two calcium-vitamin D supplementations), venous blood was drawn every 60 minutes for up to 9 hours, for serum Ca and serum PTH measurements. The order of administration of the two Ca and vitamin D supplementation sequences was allocated by randomization. No significant changes in serum Ca were observed during the study. During the 6 hours following Ca and vitamin D supplementation, a statistically significant decrease in serum PTH was observed with both regimens, compared with baseline and with the control procedure. Over this period of time, no differences were observed between the two treatment regimens. However, between the sixth and the ninth hour, serum PTH levels were still significantly decreased compared with baseline with split dose Orocal D3 administration, while they returned to baseline value with the Cacit D3 preparation. During this period, the percentage decrease in serum PTH compared with baseline was significantly more pronounced with Orocal D3 than with Cacit D3 (P = 0.0021). We therefore conclude that the administration of two doses of 500 mg of calcium and 400 IU of vitamin D3 6 hours apart provides a more prolonged decrease in serum PTH levels than the administration of the same total amount of Ca and vitamin D as a single morning dose in young healthy volunteers. This might have implications in terms of protection of the skeleton against secondary hyperparathyroidism and increased bone resorption and turnover in elderly subjects. PMID- 11870413 TI - What are the dietary requirements for calcium and vitamin D? AB - Dietary recommendations for calcium and vitamin D vary considerably among different advisory bodies. This lack of consensus reflects the different criteria used as a basis for estimating average requirements and population reference values. For calcium (Ca) many have used a factorial approach, where estimates of skeletal accretion rates are coupled with assumptions about Ca absorption and excretion on typical diets. Others have used data derived experimentally from balance studies. A recent refinement of this approach involves the mathematical modelling of balance data to estimate a Ca intake above which Ca retention is maximal or desirable. A third approach was used by a NIH Consensus Conference in 1994, based on the concept of an optimal Ca intake for bone health, in terms of reduced risk of osteoporosis in later life. Although taken up by several special interest groups, this approach has yet to be adopted fully by any committee advising on nutrition policy because of the lack of quantitative data on which to base estimates of average calcium requirements. Differences in vitamin D recommendations reflect subtle differences in philosophy about how to indicate that while endogenous synthesis is the main source of vitamin D for people with adequate sunshine exposure, there are vulnerable groups that rely on the diet to supply their vitamin D requirements. In addition, they reflect uncertainties about the relationship between vitamin D status and health outcomes. PMID- 11870414 TI - The effect of nutrient intake on bone mineral status in young adults: the Northern Ireland young hearts project. AB - Optimizing peak bone mass in early life may reduce osteoporosis risk in later life. Such optimization may be partly dependent upon diet. In the present study, nutrient intakes and selected lifestyle parameters were assessed in adolescent subjects (238 males, 205 females; aged 15 y) and again, in the same subjects, on one occasion in young adulthood (aged between 20 and 25 y). The extent of the relationships between these parameters and bone mineral density (BMD), dual energy X-ray absorptiometry (DXA), lumbar spine (L2-L4), and femoral neck measured concurrently with diet in young adulthood only, was assessed. Adjusted linear regression models were constructed. Variables included a measure of pubertal status (at age 15 y), age (at young adulthood), height, weight, physical activity, smoking, and mean daily intakes of energy, calcium, protein, vitamin D, phosphorus, total fat, and alcohol. In both sexes, body weight at adolescence and young adulthood was the only factor consistently positively associated with BMD at both measurement sites. Effects of nutrient intake on BMD were inconsistent. Vitamin D and calcium intakes reported by female adolescents showed significant positive relationships with BMD measured in young adulthood (vitamin D measured at the lumbar spine; calcium measured at the femoral neck). The positive relationship between vitamin D and BMD remained significant at young adulthood, but at the femoral neck rather than at the lumbar spine. Also in females, intakes of phosphorus and the calcium:phosphorus ratio (Ca:P) at adolescence were strongly negatively related to femoral neck BMD measured at young adulthood. In males, however, Ca:P reported at young adulthood had a significant positive relationship with lumbar spine BMD, whereas Ca:protein was negatively associated with BMD at the lumbar spine. Intakes of Ca reported by adolescent males also had a strong negative effect on lumbar spine BMD measured at young adulthood. PMID- 11870415 TI - Bisphosphonate space measurement and dosage requirement for intravenous clodronate treatment of Paget's disease. AB - It has been claimed that the bisphosphonate space, a scintigraphic technique which simultaneously estimates bone uptake and renal clearance of bisphosphonate, can be used to predict the dose of bisphosphonate required to induce biochemical remission in Paget's disease. In a prospective study of 15 newly diagnosed patients with Paget's disease, bisphosphonate space measurements were made prior to treatment with intravenous clodronate. Treatment with clodronate 0.6 g was given four times weekly until the alkaline phosphatase (ALP) suppressed below the upper level of the reference range (120 u/l) or reached a plateau, and the cumulative dose required (1.8-7.8 g) was calculated. Overall, the bisphosphonate space correlated poorly with the total dose requirement (r = 0.441, P = 0.100), but the relationship was weakened by an outlier, who had the poorest renal function. Excluding this subject improved the correlation coefficient (r = 0.852, P <0.0001). The relationship between dose requirement and log10 initial ALP was not as strong (r = 0.672, P <0.01). However, for both ALP and the bisphosphonate space the 95% prediction intervals were wide. We conclude that the bisphosphonate space does relate to dose requirements for intravenous bisphosphonates, but that it is unreliable when renal function is poor, and may not offer much gain over the pretreatment ALP levels. Both ALP and the bisphosphonate space have wide prediction intervals, and are therefore poor guides to dose requirement. PMID- 11870416 TI - Microfocus small angle X-ray scattering reveals structural features in archaeological bone samples: detection of changes in bone mineral habit and size. AB - Microfocus X-ray scattering provides a powerful nondestructive technique capable of providing important information about the size, habit, and arrangement of mineral crystals in bone. The technique is capable of probing textural differences in a sample at a micron scale resolution. The study presented here involved the analysis of a number of archaeological bones by microfocus X-ray scattering at the ESRF Grenoble in order to determine local changes in mineral durability. The results showed that regions of bone with a modified microscopic morphology contained a greater dispersion of crystal shape when compared with more intact regions and control contemporary bone samples, but the crystal thickness values showed similar consistency. We speculate that the persistence of collagen in the archaeological bone may allow diagenetic remodeling of bone in terms of crystallite shape but defines the size of remodelled crystallites. The ability to detect such local changes in texture has wide potential for determining crystal characteristics in healthy and diseased bone samples. PMID- 11870417 TI - Identification of a novel osteoblastic gene, inducible by C-type natriuretic peptide, whose transcript might function in mineralization as a noncoding RNA. AB - We reported previously that C-type natriuretic peptide (CNP) promotes the differentiation and mineralization of osteoblast-like cells. However, little information is available about the mechanism of action of CNP in differentiating osteoblastic cells. In this study, using the technique known as differential display-polymerase chain reaction, we identified a novel cDNA fragment that corresponded to a transcript whose level was increased by CNP in mouse clonal preosteoblastic MC3T3-E1 cells. Northern blotting analysis revealed transcripts of 1.3 kb and 2.3 kb in MC3T3-E1 cells. Both these transcripts were also expressed at high levels in the heart and stomach. We isolated a full-length cDNA (2,135 bp) from a cDNA library derived from MC3T3-E1 cells using the original cDNA fragment. Analysis of the sequence and of products of transcription and translation in vitro indicated that the transcript of the gene did not include any extensive open reading frames. Enhanced expression, after transfection, of transcript in MC3T3-E1 cells stimulated the deposition of calcium of these cells and the formation of mineralized nodules, but did not affect the activity of alkaline phosphatase. Our results suggest that CNP promotes the expression of a novel transcript, which might stimulate the mineralization of MC3T3-E1 cells. PMID- 11870418 TI - Cell death of osteocytes occurs in rat alveolar bone during experimental tooth movement. AB - The purpose of this study was to examine the morphological changes in alveolar bone osteocytes on the pressure side during experimental tooth movement, using quantitative evaluation on hematoxylin and eosin-stained sections, the TUNEL method, confocal laser scanning microscopy (CLSM), and transmission electron microscopy. In 8-week-old Wistar rats, the left first molar was forced to move mesially with an average load of 10 g by a nickel-titanium superelastic wire. After 6 hours, nuclear condensation and fragmentation appeared in osteocytes adjacent to the hyalinized periodontal ligament (PDL). These cells showed TUNEL positive reaction. The number of osteocytes with apoptosis progressively increased up to 1 day. At 1 and 2 days, cytoplasmic and nuclear destruction and distribution within the lacunae occurred and increased up to 4 days. The proportion of necrotic osteocytes and near empty lacunae peaked at 2 and 4 days, respectively. At 7 days, necrotic osteocyte and empty lacunae numbers returned to the level of control bone, probably due to resorption of the alveolar bone containing apoptotic and necrotic osteocytes. Ultrastructually, the osteocytes showed apoptotic morphology at 6 and 12 hours and 1 day; at 2 and 4 days, several osteocytes exhibited characteristics of necrosis and destructive images of the surrounding bone matrix, which resulted in enlargement of the lacunae. The present results demonstrate that osteocytes in alveolar bone adjacent to the hyalinized PDL underwent cell death via apoptosis and "secondary necrosis" during orthodontic tooth movement, which may be associated with the subsequent bone resorption. PMID- 11870421 TI - The role of MRI in pancreatic cancer. PMID- 11870419 TI - Localization of cathepsin K in bovine odontoclasts during deciduous tooth resorption. AB - Cathepsin K is a cysteine proteinase, which is abundantly and selectively expressed in osteoclasts. It is believed to play an important role in the proteolysis of bone resorption by osteoclasts. The objectives of this study were to investigate the association of cathepsin K in the physiological root resorption of deciduous teeth and to identify the cathepsin K-producing cells in deciduous root resorption. RT-PCR and Northern blot analysis of the total RNAs extracted from bovine active and resting root-resorbing tissues, which lie between the root of deciduous tooth and its permanent successor, were performed. The active root-resorbing tissue, which has a high population of odontoclasts on its surface that is attached to resorbing root surface, showed an extremely high expression of cathepsin K in comparison with the resting root-resorbing tissue. By in situ hybridization, cathepsin K mRNA was highly and selectively expressed in multinucleated odontoclasts that aligned along the surface of the tissue and apposed to the resorbing root surface of the deciduous tooth. Western blot analysis of the active root-resorbing tissue was used to characterize the anti cathepsin K antibody. A band of 27 kDa, corresponding with the predicted size for mature cathepsin K, was demonstrated. Immunohistochemistry confirmed the specific localization of cathepsin K protein to the odontoclasts. These results demonstrate that odontoclasts in the deciduous root resorption express cathepsin K mRNA and protein that may participate in the proteolysis of root resorption of the deciduous tooth. PMID- 11870422 TI - Ultrasound of the gallbladder and the biliary tree. PMID- 11870423 TI - Interventional ultrasound. AB - Modern cross-sectional imaging methods of computed tomography, sonography and lately MRI have contributed greatly to non-invasive studies of different parenchymal organs and permit the evaluation of pathological changes in different areas of the body. Although these techniques have reached a very high level of refinement, it is often not possible to define whether a focal lesion is benign or malignant. Ultrasound-guided fine-needle aspiration biopsy (FNA) or core needle biopsy (including high-speed- and vacuum-assisted techniques) are essential to find out the character of the disease in these cases. Experience of more than 10,000 interventions of different organs are presented. Technique, results, problems, as well as risks are discussed. Moreover, it is shown how the results of percutaneous biopsies often change the treatment of a patient. This is especially of interest in those suffering from lymphomas, pancreatic carcinomas, and breast cancers, as well as relapsing tumors. PMID- 11870424 TI - Ultrasound-guided drainages and sclerotherapy. AB - Ultrasonically guided fluid collection and abscess drainage have become routine procedures in various parts of the body. In most cases ultrasound is the only imaging and guidance modality needed; however, it is of the utmost importance to remember that CT and fluoroscopy with contrast often give invaluable information when the true extent of the process has to be determined and when assessing the safest route for the catheter in anatomically complicated areas. The importance of irrigation of the abscess cavity with fluids and the ready use of urokinase should be emphasized. Ethanol sclerotherapy is a simple and safe procedure to treat symptomatic hepatic or renal cysts. Parathyroid adenomas and cysts, as well as thyroid cysts, can also be treated with ethanol sclerotherapy in selected cases. Purified mineral talc has been used in pleurodesis and hydrocele sclerotherapy, whereas doxycycline or ethanol is used for postoperative lymphoceles. Both abscess drainages and sclerotherapy procedures are minimally invasive, simple, safe, inexpensive and reasonably efficacious treatment in many clinical instances and may be at least an alternative to surgical treatment, often offering significant advantages over surgery. PMID- 11870425 TI - Preoperative local MRI-staging of patients with a suspected pancreatic mass. AB - The aim of this study was to define the value of MRI of the pancreas for preoperative local staging of patients with a suspected pancreatic mass. Ninety four patients (41 women, 53 men; age range 32-87 years) with a suspected pancreatic tumor underwent preoperative staging with MRI on a 1.5-T system. The MRI protocol included breath-hold MR cholangiopancreatography in turbo spin-echo technique, biphasic contrast-enhanced 3D MR angiography, and MRI of the upper abdomen with breath-hold T2-weighted half-Fourier acquired single-shot turbo spin echo and T1-weighted fast-low-angle-shot (pre- and postcontrast) sequences. Data were collected prospectively and analyzed by two radiologists in agreement modality. Evaluation criteria were vascular involvement, resectability, and a characterization benign vs malignant. Results were compared to histopathology in 78 patients. Sixteen patients were followed-up. In 74 of 94 patients a solid tumor or an inflammation of the pancreas ( n=62) or the papilla ( n=12) was detected. In this group, MRI had a sensitivity of 98%, a specificity of 92%, and an accuracy of 96% in the characterization of malignant tumors. Regarding only the solid tumors, the positive predictive value of MRI was 87% with respect to resectability. Other pathologic findings included adenoma or inflammation of the duodenum ( n=5), carcinoma or benign stenosis of the choledochus duct ( n=7) and carcinoma of the gall bladder ( n=2). In 6 patients MRI did not depict any pathologic findings, and follow-up confirmed this interpretation. Magnetic resonance imaging allows a local preoperative staging in patients with suspected pancreatic tumor. Limitations, however, concern to the diagnostics of peritoneal and/or liver metastases. PMID- 11870426 TI - Multifocal inflammatory pseudotumor of the liver: dynamic gadolinium-enhanced, ferumoxides-enhanced, and mangafodipir trisodium-enhanced MR imaging findings. AB - The MRI characteristics of a multifocal inflammatory pseudotumor of the liver are described. Emphasis is placed on the appearances following intravenous administration of both non-specific and liver-specific MR contrast agents. On post-gadolinium gradient-echo (GE) images an early, intense, and peripheral enhancement was followed by a homogeneous, complete, and persistent enhancement. Lesions showed no uptake following administration of ferumoxides particles nor mangafodipir trisodium, respectively. During follow-up, a peripheral hyperintense rim appeared on precontrast T1-weighted images, a feature not previously described. PMID- 11870427 TI - The value of MR angiography in predicting the risk of torsion of a pelvic spleen during pregnancy. AB - A case of an enlarged pelvic spleen, studied with MRI and MR angiography (MRA), is presented in a 32-year-old female wishing to become pregnant. An ectopic located spleen may be complicated by an acute abdomen due to torsion of the splenic vascular pedicle, resulting in splenic infarction. Displacement of the spleen and splenic pedicle during pregnancy may further increase the risk of torsion. Urgent splenectomy during pregnancy is associated with a high fetal and maternal mortality and morbidity. On the other hand, elective splenectomy of a pelvic spleen before pregnancy can result in adhesion formation, compromising the patient's fertility. The abilities of MRI and MRA in predicting the risk of these life-threatening complications during pregnancy are discussed, in order to evaluate the benefit-risk ratio of surgical treatment by splenectomy of splenopexia. PMID- 11870428 TI - Imaging of abdominal tuberculosis. AB - The concept of "abdominal tuberculosis" in this review refers to peritoneum and its reflections, gastrointestinal tract, abdominal lymphatic system, and solid visceral organs, as they are subject to varying degrees of involvement alone or in combination. Some features, including free or loculated ascites with thin mobile septa, smooth peritoneal thickening and enhancement, misty mesentery with large lymph nodes, smudged omental involvement, and advanced ileocecal changes demonstrated by US, CT, or gastrointestinal series are deemed suggestive radiological findings. The diagnosis still requires a high index of suspicion, once the suggestive features have been demonstrated by imaging modalities. PMID- 11870429 TI - US and CT diagnosis of complete cecocolic intussusception caused by an appendiceal mucocele. AB - Appendiceal mucoceles are found in only 0.2-0.3% of all appendectomy materials. Colocolic intussusception of the appendix is also very uncommon. We report the very rare association of these two entities in a 40-year-old patient presenting with intermittent right abdominal pain accompanied by a palpable mass in the right flank. The full diagnosis was made preoperatively by ultrasound and confirmed by helical CT by means of unequivocal signs of intussusception associated with a very suggestive "cup-and-ball" aspect of the mucocele induced by a global mucinous cystadenoma of the appendix. A brief review of the available literature on mucocele is given. PMID- 11870430 TI - Radiographic findings in 37 cases of primary CNS lymphoma in immunocompetent patients. AB - Because of the increasing incidence of primary central nervous system lymphoma (PCNSL), it is essential to recognize this disease in order to start appropriate treatment. We present the characteristic CT and MRI features of this tumour. The findings of 32 CT and 31 MR of 37 immunocompetent patients with biopsy-proved PCNSL are reviewed. The main features are presented and analysed, and are discussed in comparison with proven literature data. Primary central nervous system lymphoma presents as supratentorial solitary lesions in approximately 80% of the patients and multiple lesions in 20%. In contrast to classical data, the lesions are located in deep structures only in one-third of the cases, and involve posterior fossa in 10% of cases. Most of the lesions are hyperdense or isodense (92%) on CT, hypointense or isointense on T1-weighted images, and only about 40% are hyperintense on T2-weighted images. Nearly all the lesions enhance, except after corticosteroid administration. They produce mild oedema and mass effect. Meningeal or ventricular enhancement are rare but suggestive. Calcification, haemorrhage or necrosis are scarce. Although PCNSL in immunocompetent patients have a variable CT and MR appearance, the imaging data often suggest the diagnosis. PMID- 11870431 TI - Tolosa-Hunt syndrome in a patient with systemic lupus erythematosus. AB - We report a case of Tolosa-Hunt syndrome (THS) in a patient with systemic lupus erythematosus studied with MRI. Magnetic resonance showed enlargement of the cavernous sinus and compression of the carotid syphon by enhancing tissue. In particular, fat-suppressed T1-weighted images before and after contrast agent injection and MR angiography showed extension of the abnormal tissue to the apex of the orbit and narrowing of the internal carotid artery. A presumptive diagnosis of THS was made and steroid treatment was started with rapid relief of symptoms. Follow-up MR study after steroid therapy demonstrated sub-total resolution of the neuroradiological findings. Neuroradiological findings in THS are quite typical but they may be subtle; furthermore, the presence of a systemic disease may suggest secondary involvement of the cavernous sinus. Utilization of the appropriate MR techniques and follow-up exams may contribute to the diagnosis of THS even in the presence of other systemic diseases. PMID- 11870432 TI - Therapy-related change of corpus callosum in a young patient with epilepsy. AB - Focal nonhemorrhagic lesion in the splenium of the corpus callosum in a patient with epilepsy treated with antiepileptic drugs was observed with MRI imaging. We have found only one such case during the past 2 years (series of MRI examinations of approximately 500 patients with various forms of epilepsy). PMID- 11870433 TI - Imaging of MALT lymphomas. AB - The broad category of non-Hodgkin's lymphoma includes a large variety of different diseases including indolent as well as aggressive lymphomas. Mucosa associated lymphoid tissue (MALT) lymphoma arises in the extranodal mucosal lymphoid tissue and has only been recognised as a distinct entity in recent years. It affects one or several extranodal structures such as the stomach, the lung, the eye and salivary glands. The lymphoma is generally of low grade and has indolent course. The aim of this article is to exemplify the most common radiological patterns of MALT lymphoma. PMID- 11870434 TI - Characterization of low-intensity lesions in the peripheral zone of prostate on pre-biopsy endorectal coil MR imaging. AB - The aim of this study was to determine which morphological features of low intensity lesions in the peripheral zone of the prostate are predictable of prostate cancer on pre-biopsy T2-weighted integrated endorectal phased-array MR images. The MR examinations were performed in 69 consecutive patients with elevated level of prostate-specific antigen (>4 ng/ml) and/or a positive digital rectal examination before transperineal 12-site biopsy. Two radiologists evaluated presence of lesions, their morphological features, and possibility of malignancy in divided into four sections of the peripheral zone. Imaging analysis findings were compared with biopsy results. Discriminative features were selected by stepwise logistic regression. Descriptive statistics and receiver operating characteristics (ROC) curves were also calculated. Sixty-eight benign lesions and 23 malignant lesions were found. Wedge shape and diffuse extensions without mass effect were significantly associated with benignity ( P=0.0105 and 0.002, respectively). Lesion size was significantly associated with malignancy ( P=0.0001). For evaluating probability of malignancy for lesions, regression model showed a comparable accuracy with the total impression for the readers in ROC analysis (Az 0.9095 vs 0.9266, respectively). Wedge shape, diffuse extension without mass effect, and size are the morphological features of low-intensity lesions in the peripheral zone on pre-biopsy T2-weighted MR images that give the best prediction of malignancy. PMID- 11870435 TI - Investigation of vesicoureteral reflux with colour Doppler sonography in adult patients with spinal cord injury. AB - The aim of the study was to evaluate prospectively the performances of colour Doppler sonography for detection of vesicoureteral reflux in adult patients with neurogenic bladder due to spinal cord injury. One hundred eighty-seven adult patients who had developed neurogenic bladder due to spinal cord injury underwent retrograde cystography and sonographic evaluation by B-mode and colour flow Doppler sonography (CFDS). Results of both examinations were analyzed independently by two radiologists. Representative images of the sonographic evaluations were recorded on videotape and CD-Rom. CFDS detected reflux in 20 ureters while retrograde cystography revealed vesicoureteral reflux (VUR) in 25 ureters of a total of 374 ureters, which were examined by both methods. Sonography detected all cases of grades IV and V of reflux, 7 of 8 (87.5%) of grade III, 5 of 6 (83.3%) of grade II, and 4 of 7 (57.14%) of grade I. There were 6 false-positive and 5 false-negative findings with Doppler examination compared with retrograde cystography. Therefore, sensitivity and specificity of sonographic examination were 80 and 98.28%, respectively, with a positive predictive value of 76.92%. Colour flow Doppler sonography can play an important role in detection of VUR in spinally injured adult patients with neurogenic bladder and can be an effective imaging tool for follow-up. PMID- 11870436 TI - Burned-out tumor of the testis presenting as supraclavicular lymphadenopathy. AB - Burned-out tumor of the testis is a rare clinical entity. It generally presents with metastases and is nonpalpable in testicular palpation. We present a case of testicular burned-out tumor having supraclavicular and retroperitoneal lymph node metastases. Imaging findings of such tumors have insufficiently been documented in radiology literature. Scrotal sonography is crucial in detecting the regressed tumors especially in patients with extragonadal metastasis of a testicular primary. PMID- 11870437 TI - Male splenic-gonadal fusion of the continuous type: sonographic findings. AB - A new case of splenic-gonadal fusion in an 8-year-old boy is reported in which the sonographic findings were highly suggestive to include the condition in the differential diagnosis. A separate mass with medium-level echoes and a homogeneous echo texture, surrounded by a clearly visible echogenic capsule, was found in contact with the lower pole of the left testis. The particular character of this congenital anomaly is emphasized and the pathogenesis, diagnosis, and treatment are discussed. PMID- 11870438 TI - Analysis of artefacts and detail resolution of lung MRI with breath-hold T1 weighted gradient-echo and T2-weighted fast spin-echo sequences with respiratory triggering. AB - The aim of this study was to evaluate feasibility and limitations of two MR sequences for imaging of the lung using a semi-quantitative rating scale. Ten healthy volunteers were assessed with a breath-hold T1-weighted gradient-recalled echo (TR/TE=129/2.2 ms, matrix 173 x 256) and a T2-weighted turbo spin-echo (TSE) sequence with respiratory triggering (TR/TE=3000-5000/120 ms, matrix 270 x 512) in axial 6-mm slices. The T1-weighted GRE protocol included a pre-saturation pulse over the mediastinal structures. Artefacts and resolution of vessel/airway structures in each lung segment were evaluated by two observers (10 volunteers, 180 segments). Cardiac and vessel pulsation artefacts predominated on T1-weighted GRE, respiration artefacts on T2-weighted TSE (lingula and middle lobe). Pre saturation of the mediastinum reduced pulsation artefacts on T1-weighted GRE. T1 weighted GRE images were improved by bright flow signal of vessels, whereas image quality of T2-weighted TSE was reduced by black-blood effects in central parts of the lung. Delineation of lung periphery and the mediastinum was superior with T2 weighted TSE. Segmental/sub-segmental vessels (up to fourth/fifth order) and bronchi (up to third order) were identified. All 180 lung segments were imaged in diagnostic quality with at least one of the two sequences (T1-weighted GRE not diagnostic in 9 of 180, T2-weighted TSE in 4 of 180). Both sequences were found to be complementary: superior identification of gross lung anatomy with T1 weighted GRE and higher detail resolution in the periphery and the mediastinum with T2--weighted TSE. PMID- 11870439 TI - Chest imaging with flat-panel detector at low and standard doses: comparison with storage phosphor technology in normal patients. AB - The purpose of this study was to evaluate the performance of standard-dose and low-dose cesium iodide (CsI)-doted amorphous silicon (a-Si) flat-panel detector technology (FDT) as compared with storage-phosphor technology (SPT) in the depiction of relevant anatomical structures in chest radiography. In 75 patients referred for thoracic CT, digital chest radiographs were randomly obtained with either SPT at a standard dose (speed class S400, n=25), standard-dose FDT (S400, n=25) or FDT at a low dose (S800, n=25). Five radiologists evaluated the visibility of eight pulmonary and mediastinal anatomical structures using a five point rating scale. To determine statistically significant differences between the three groups, the Mann-Whitney U-test was employed. No statistically significant differences were found in the depiction of eight criteria between SPT and standard-dose or low-dose FDT chest radiographs. The performance of FDT S400 was equal to SPT for most criteria and better for retrocardiac structures and soft tissue. FDT S800 was inferior to both SPT and FDT S400. Standard-dose FDT is equivalent to SPT in the depiction of relevant anatomical structures of the chest. Our results also indicate that a dose reduction of 50% with FDT may result in small but not significant decrease of image quality. PMID- 11870440 TI - Imaging of cavitary necrosis in complicated childhood pneumonia. AB - The aim of this study was to illustrate the chest radiographs (CR) and CT imaging features and sequential findings of cavitary necrosis in complicated childhood pneumonia. Among 30 children admitted in the Pediatric Intensive Care Unit for persistent or progressive pneumonia, respiratory distress or sepsis despite adequate antibiotic therapy, a study group of 9 children (5 girls and 4 boys; mean age 4 years) who had the radiographic features and CT criteria for cavitary necrosis complicated pneumonia was identified. The pathogens identified were Streptococcus pneumoniae( n=4), Aspergillus( n=2), Legionella( n=1), and Staphylococcus aureus( n=1). Sequential CR and CT scans were retrospectively reviewed. Follow-up CR and CT were evaluated for persistent abnormalities. Chest radiographs showed consolidations in 8 of the 9 patients. On CT examination, cavitary necrosis was localized to 1 lobe in 2 patients and 7 patients showed multilobar or bilateral areas of cavitary necrosis. In 3 patients of 9, the cavitary necrosis was initially shown on CT and visualization by CR was delayed by a time span varying from 5 to 9 days. In all patients with cavities, a mean number of five cavities were seen on antero-posterior CR, contrasting with the multiple cavities seen on CT. Parapneumonic effusions were shown by CR in 3 patients and in 5 patients by CT. Bronchopleural fistulae were demonstrated by CT alone ( n=3). No purulent pericarditis was demonstrated. The CT scan displayed persistent residual pneumatoceles of the left lower lobe in 2 patients. Computed tomography is able to define a more specific pattern of abnormalities than conventional CR in children with necrotizing pneumonia and allows an earlier diagnosis of this rapidly progressing condition. Lung necrosis and cavitation may also be associated with Aspergillus or Legionella pneumonia in the pediatric population. PMID- 11870441 TI - Spectrum of abdominal sonographic findings in 103 pediatric patients with Gaucher disease. AB - The purpose of this study was to catalogue spleen and liver features by US in a cohort of 103 pediatric patients with Gaucher disease, and to document response to enzyme replacement therapy by serial US examination. There were 42 boys and 61 girls, 2 were splenectomized. Forty-eight patients received enzyme replacement therapy (ERT). At presentation all patients evinced organomegaly and 4.9% had focal (splenic or hepatic) lesions (hypo- and hyper-echoic or mixed). Fifteen patients began ERT before 11 years of age. There was a mean liver volume reduction from 230 to 137% after 2 years of ERT, with further reduction of 91% up to 6 years later. Mean spleen volume reduction was 38.4% at 2 years of ERT and a further reduction to 40.8% at last evaluation. Fourteen children began ERT between ages 11 and 16 years. Mean liver reduction was from 230 to 124% at 2 years and further reduction to mean of 104% recently. Mean splenic reduction was 37.7% after 2 years of ERT, with a mean of 43.8% recently (mean 4.5 years later). Organ volume changes in untreated and treated children were documented by US which is the modality of choice for repeat measures in this population. Our results highlight the initial dramatic and then continued response to ERT in pediatric patients with Gaucher disease. PMID- 11870442 TI - Bone structure of the distal radius and the calcaneus vs BMD of the spine and proximal femur in the prediction of osteoporotic spine fractures. AB - The aim of this study was to compare structure measures obtained from high resolution MR images of the calcaneus and the distal radius with bone mineral density (BMD) of the spine and hip in the prediction of osteoporotic spine fracture status. High-resolution MR images of the calcaneus and radius were obtained in 24 post-menopausal women with spine fractures and 22 age-matched controls. Imaging was performed at 1.5 T using a T1-weighted spin-echo sequence (slice thickness 1 mm, in-plane spatial resolution 195 x 195 microm(2)). Structure analysis was performed using parameters analogous to standard histomorphometry. Bone mineral density of the spine was obtained using quantitative CT and of the hip with dual-energy X-ray absorptiometry. Significant differences between both patient groups were obtained with BMD and all structure parameters ( p<0.05). Using receiver operating characteristic analysis to determine the diagnostic performance in differentiating both groups, the best results were found for BMD of the spine, one of the radial structure measures and a combination of the calcaneal structure measures. In this study BMD of the spine and structure measures of the distal radius were best suited to predict the osteoporotic fracture status of the spine. A combination of BMD and structure measures did not yield any additional information on fracture status. PMID- 11870443 TI - Imaging features of iliopsoas bursitis. AB - The aim of this study was firstly to describe the spectrum of imaging findings seen in iliopsoas bursitis, and secondly to compare cross-sectional imaging techniques in the demonstration of the extent, size and appearance of the iliopsoas bursitis as referenced by surgery. Imaging studies of 18 patients (13 women, 5 men; mean age 53 years) with surgically proven iliopsoas bursitis were reviewed. All patients received conventional radiographs of the pelvis and hip, US and MR imaging of the hip. The CT was performed in 5 of the 18 patients. Ultrasound, CT and MR all demonstrated enlarged iliopsoas bursae. The bursal wall was thin and well defined in 83% and thickened in 17% of all cases. The two cases with septations on US were not seen by CT and MRI. A communication between the bursa and the hip joint was seen, and surgically verified, in all 18 patients by MR imaging, whereas US and CT failed to demonstrate it in 44 and 40% of the cases, respectively. Hip joint effusion was seen and verified by surgery in 16 patients by MRI, whereas CT (4 of 5) and US ( n=12) underestimated the number. The overall size of the bursa corresponded best between MRI and surgery, whereas CT and US tended to underestimate the size. Contrast enhancement of the bursal wall was seen in all cases. The imaging characteristics of iliopsoas bursitis are a well-defined, thin-walled cystic mass with a communication to the hip joint and peripheral contrast enhancement. The most accurate way to assess iliopsoas bursitis is with MR imaging; thus, it should be used for accurate therapy planning and follow-up studies. In order to initially prove an iliopsoas bursitis, US is the most cost-effective, easy-to-perform and fast alternative. PMID- 11870444 TI - MR imaging of the foot and ankle: patterns of bone marrow signal abnormalities. AB - Diagnosis of marrow disorders of the foot and ankle is among the more challenging aspects of MR interpretation. Evaluation of normal and abnormal bone marrow with regard to pattern, distribution, and signal characteristics on different sequences often allows a specific diagnosis. This pictorial review illustrates MR imaging findings of normal variants of bone marrow of the foot and ankle, and the varied responses of bone marrow to trauma, stress, or disease. PMID- 11870445 TI - Clear cell chondrosarcoma: unusual radiologic appearances with histologic correlation. AB - Clear cell chondrosarcoma is a rare variant of the bone tumors with distinct radiologic and pathologic features. In this report two cases of clear cell chondrosarcomas with atypical radiologic appearances are presented. The findings of X-ray films and MRI are described with histologic correlation. In the first case the lesion showed a very expansile and long segment involvement of the humerus. In the second case the lesion was located in the diaphysis of the femur causing a large cortical destruction. PMID- 11870446 TI - Imaging of acute traumatic injuries of the thoracic aorta. AB - Blunt traumatic aortic injuries are a major concern in the settings of high-speed deceleration accidents, since they are associated with a very high mortality rate; however, with prompt diagnosis and surgery, 70% of the patients with a blunt aortic lesion who reach the hospital alive will survive. This statement challenges the emergency radiologist in charge to evaluate the admission radiological survey in a severe chest trauma patient. With a 95% negative predictive value for the identification of blunt traumatic aortic lesions, plain chest film represents an adequate screening test. If aortography remains the gold standard, it tends, at least in hemodynamically stable trauma patients, to be replaced by spiral-CT angiography (SCTA), which demonstrates a 96.2% sensitivity, a 99.8% specificity, and a 99.7% accuracy. In unstable patients, trans-esophageal echography (TEE) plays a major diagnostic role. Knowledge of advantages and pitfalls of these imaging techniques, as reviewed in this article, will help the emergency radiologist to choose the appropriate algorithm in the diagnosis of traumatic aortic injury, for each trauma patient. PMID- 11870447 TI - Catheter-tip fixation of a percutaneously implanted port-catheter system to prevent dislocation. AB - The aim of this study was to evaluate the incidence of catheter tip dislocation in patients with percutaneously implanted port-catheters for hepatic arterial chemotherapy with catheter tip fixation. Forty-seven patients (31 men and 16 women; mean age 66 years) with unresectable advanced liver cancers (primary liver cancer, n=19; metastatic liver cancer, n=28) underwent percutaneously implantable port-catheter system placement with the tip fixed at the gastroduodenal artery with coils and side hole opened at the common hepatic artery. In 39 patients, n butyl cyanoacrylate (NBCA) mixed with Lipiodol was added for fixation. The position of the side hole after the indwelling port-catheter system was investigated, and the correction method in cases with catheter dislocation was determined. In 2 (25%) of the 8 patients without NBCA fixation, dislocation of the catheter was noted, in contrast to none (0%) of 37 patients with NBCA fixation. Two patients in whom NBCA was used could not undergo long-term intra arterial chemotherapy because of hepatic arterial thrombotic occlusion which occurred after placement of the indwelling catheter, and were excluded from the evaluation. Fixation of the catheter tip with combined use of coils and NBCA- Lipiodol mixture to the gastroduodenal artery is important to prevent dislocation of the port-catheter system. PMID- 11870449 TI - Transcatheter coil embolization of an intercostal artery to pulmonary artery fistula. AB - An 18-year-old asymptomatic male was found to have a high-flow systemic arterial to pulmonary arterial fistula fed by dilated lower intercostal arteries draining via pulmonary artery toward the left main pulmonary trunk. Transcatheter Guglielmi Detachable Coil (GDC; Target Ther, Fremont, Calif.) embolization was performed. Transcatheter embolization is a reasonable and less invasive mode in the treatment of systemic artery to pulmonary artery fistula, and GDC offers more precise coil placement over other conventional coils. PMID- 11870448 TI - Pseudoaneurysm and ilio-caval fistula caused by malignant fibrous histiocytoma of the aorta--CT diagnosis and angiographic confirmation. AB - We report a case of a malignant fibrous histiocytoma (MFH) of the aortic bifurcation, which manifested as a pseudoaneurysm with the formation of an ilio caval fistula, a complication about which, to our knowledge, nothing has been published previously. Spiral CT, catheter arteriography and venography were complementary in the diagnostic procedure. PMID- 11870450 TI - Tracheobronchial stent insertions in the management of major airway obstruction in a patient with Hunter syndrome (type-II mucopolysaccharidosis). AB - We report a case of a 22-year-old male with Hunter syndrome who developed progressive major airway obstruction and was treated with insertion of plastic and metallic stents, with dramatic improvement in the patient's symptomatic and functional status. To the best of our knowledge, this is the first reported case of endoluminal stents being used in the management of major airway obstruction in a patient with Hunter syndrome. PMID- 11870452 TI - Clip placement facilitating the approach to breast lesions. AB - We describe a novel technique for marking non-palpable breast lesions with the aim of selecting the best approach prior to performing a biopsy. The technique employs a new coil, specifically designed for breast localization, guided by stereotaxy. This technique is reserved for selected cases in which the lesion is seen peripherally in only one mammographic view with negative or non-conclusive ultrasonographic results, and deeply seated after a stereotactic study. Once the coil is released beside the lesion, the shortest approach from the skin may be employed to perform the biopsy. To our knowledge, this is the first report of this technique. PMID- 11870451 TI - Quality control of the physical and technical aspects of mammography in the Norwegian breast-screening programme. AB - The aim of this study was to establish a system for quality control of technical and physical aspects of mammography in the national Norwegian breast-screening programme, and to evaluate test results obtained during the 4-year pilot project period. Quality control (QC) procedures for the X-ray unit, film and processing, cassettes and screens, view boxes, and image quality were described in two QC manuals. Frequent and relatively simple tests were performed by radiographers on site and results reported semi-annually to an inspection group established at the Norwegian Radiation Protection Authority (NRPA). Tests requiring special equipment were performed prior to start of screening (first status controls), then annually by the NRPA group, which also assembled and analysed the data from the first 4 years. The analyses were done mainly with reference to stated standard or guidance levels and the degree of deviation from such values. Results for tests of i.a. focal spot size, exposure time, radiation output, mean absorbed glandular dose, reference optical density, sensitometry, and AEC performance are presented. The standard of the equipment tested was in general high. Although some deviations from optimal performance were seen throughout the project, the equipment mostly performed well within the stated limiting values. PMID- 11870454 TI - Intranet and radiology: a critical appraisal of radiological applications of Intranet technology. AB - The World-Wide Web (WWW or Web) is the service which led to the huge popularity of the Internet by making it user friendly. Already in the early years of the Web, this technology was also used to make internal information systems easier to use and hospitals and departments set up "Intranets". An Intranet consists of a Web Server which is installed within a local area network (LAN) and allows information retrieval with a Web browser. This paper highlights the different fields of Intranet applications in radiology and in the hospital and focuses on systems for organisational issues as well as for patient data distribution. While an Intranet can be a solution for many problems, not only in radiology, it is accompanied by serious threats and common misunderstandings which are discussed. PMID- 11870453 TI - Contrast media as markers of GFR. AB - Determination of the glomerular filtration rate (GFR) is generally considered as the most important parameter of quantifying renal function. The GFR is determined as renal or plasma clearance of an ideal filtration marker which is freely filtered by the kidney, does not undergo metabolism, tubular secretion or absorption. Markers that fulfil these demands are inulin, 51Cr-EDTA, 99mTc-DTPA, labelled or unlabelled contrast media. The renal clearance of inulin is the classic reference method for estimation of the GFR. This method is however not practical for routine clinical purposes. Radionucleids have therefore been used as alternative filtration markers since the 60s. Drawbacks related to radiation exposure especially in children and pregnant women and the safety in handling radiolabelled markers have led to an increasing interest in using non-radioactive markers. The development of simple and reliable methods to determine the concentration of contrast media in plasma and urine, such as high-performance liquid chromatography (HPLC) and X-ray fluorescence analysis have made this possible. The non-ionic low osmolar contrast medium iohexol has become the most commonly used contrast medium for GFR measurements in Europe. However, other contrast media with similar pharmacokinetics may be equally suitable as GFR markers. PMID- 11870455 TI - Comments on Zanetti et al.: MR arthrographic variability of the arthroscopically normal glenoid labrum: qualitative and quantitative assessment. PMID- 11870462 TI - Role of MR imaging in epilepsy. PMID- 11870463 TI - Colonography: current status, research directions and challenges. Update 2002. AB - With the introduction of multislice CT technology and faster gradients in MRI, CT and MR colonography are increasingly becoming attractive alternatives for colorectal mass screening. In this article we summarise the current status, research directions, and challenges in CT and MR colonography. PMID- 11870464 TI - CT colonography: colonic distention improved by dual positioning but not intravenous glucagon. AB - The aim of this study was to determine whether intravenous (IV) glucagon and dual positioning administered prior to CT colonography enhances colonic distention. We assessed the effect of dual positioning and IV glucagon on colonic distention in 96 patients who underwent CT colonography examinations. The CT colonography was performed in both supine and prone positions. Seventy-four patients received glucagon (1 mg i.v.) immediately prior to CT scanning and 22 patients did not. The bowel was divided into ten segments and colonic distention was scored by two radiologists in the supine, prone, and combined supine/prone positions using a five-point scale: 1=collapsed; 2=poorly visualized; > or = 3=adequate distention; 4=entire segment visualized and well distended; 5=excellent distention). A combined segmental and overall supine/prone distention score was calculated based on the sum of the mean score for each position. There was no significant difference in the degree of colonic distention between patients who received glucagon and those who did not [supine/prone distention score (mean +/- SE): 3.63 +/- 0.2 vs 3.85 +/- 0.2; p=n.s.]. The degree of colonic distention was greater in the prone position in both the glucagon (3.87 +/- 0.2 vs 3.38 +/- 0.2; p<0.05) and non-glucagon groups (4.01 +/- 0.2 vs 3.69 +/- 0.2; p=N.S.) particularly in the proximal colon. There was almost perfect agreement between both radiologists in their scoring of colonic distention on a per-patient basis ( k=0.9; p<0.001). Of 1480 bowel segments, 1261 (85.2%) were adequately distended in the glucagon group compared with 370 of 440 bowel segments (84%) in the non-glucagon group ( p=n.s.) Colonic distention at CT colonography is improved by dual positioning but not by the administration of intravenous glucagon. While our results suggest that other smooth muscle relaxants, including butyl scopolamine, may only have a limited role in improving colonic distention in CT colonography, further studies are required. PMID- 11870465 TI - An unusual case of herniation of small bowel through an iatrogenic defect of the falciform ligament. AB - Internal herniation through a congenital defect in the falciform ligament is extremely rare. We report an unusual observation of small bowel obstruction through an iatrogenic defect of the falciform ligament. Prompt diagnosis was made by helical CT, permitting a rapid surgical procedure to preserve the viability of the obstructed segment. PMID- 11870466 TI - Small bowel intussusception caused by intestinal angiosarcomatosis: usefulness of MR enteroclysis with infusion of water through a nasojejunal catheter. AB - We report a case of intestinal angiosarcomatosis manifesting as jejuno-jejunal intussusception. PMID- 11870467 TI - Characterization of liver hemangiomas with pulse inversion harmonic imaging. AB - The purpose of this study was to determine if pulse inversion harmonic imaging (PIHI) can characterize liver hemangiomas. We retrospectively evaluated 39 consecutive patients with liver hemangiomas, 20 typical on conventional US (hyperechoic, homogeneous, or slightly inhomogeneous and with sharp margins) and 19 atypical (11 inhomogeneous with different echogenicity larger than 3 cm, 6 hypoechoic, and 2 isoechoic smaller than 3 cm). Each liver hemangioma was firstly evaluated by PIHI and then confirmed by dynamic helical CT (28 patients) or by 6 months of US follow-up (11 patients). The PIHI was performed by two distinct sweeps on a marker lesion, 30 s (vascular phase) and from 3 to 5 min (late phase) after bolus injection of Levovist (2.5 g, 300 mg/ml). Scans were digitally stored and reviewed using a dedicated software. Contrast enhancement features of marker lesion were subjectively evaluated. Typical hemangiomas on conventional US revealed on PIHI a characteristic rim-like or peripheral globular enhancement on 30-s scan in 4 of 20 cases (20%) and a characteristic isoechoic pattern on late phase in 16 of 20 cases (80%). On PIHI, all (11 of 11) atypical hemangiomas larger than 3 cm and 4 of 8 atypical liver hemangiomas smaller than 3 cm revealed a characteristic rim-like or peripheral globular enhancement on vascular phase with a characteristic centripetal fill-in on late phase. In 4 of 8 atypical liver hemangiomas smaller than 3 cm no characteristic pattern was revealed by PIHI. Pulse inversion harmonic imaging revealed a typical pattern in the majority of liver hemangiomas typical and atypical on conventional US. In few liver hemangiomas atypical on conventional US PIHI did not identify a characteristic pattern and helical CT was necessary for final characterization. PMID- 11870468 TI - Imaging findings in the rare catastrophic variant of the primary antiphospholipid syndrome. AB - We report imaging findings in a case of the rare catastrophic variant of antiphospholipid syndrome (CAPS) characterized by widespread microvascular occlusions, which may lead to multiple organ failure. We present a case of a 66 year-old woman with bone marrow necrosis, acute acalculous cholecystitis (AAC), focal liver necrosis, subtle patchy splenic infarctions, and bilateral adrenal infarction. The demonstration of multiple microvascular organ involvement (three or more) is crucial for the diagnosis of the catastrophic variant of APS. This can be performed radiologically intra-vitam. Imaging can even reveal subclinical microinfarctions, which are often only diagnosed at autopsy. PMID- 11870469 TI - MR imaging in epilepsy that is refractory to medical therapy. AB - The aim of this study was the assessment of detection rate on MRI and description of MRI findings in patients with medically intractable epilepsy. Seventy-three patients with medically intractable epilepsy between the ages of 0 and 68 years old were evaluated by MRI, on three planes with spin-echo T1, fast spin-echo T2, and fluid-attenuated inversion recovery sequences, and, if necessary, with contrast-enhanced SE T1 sequences. Cerebral infarct regions with atrophy and gliosis in 8 patients, cerebral tumors in 5 patients, hippocampal sclerosis in 16 patients, radial microbrain in 1 patient, cortical dysplasia in 3 patients, pachygyria in 2 patients, subcortical heterotopia in 2 patients, schizencephaly in 3 patients, cerebral hemiatrophy in 2 patients, tuberous sclerosis in 1 patient, herpes encephalitis in 2 patients, Rasmussen's encephalitis in 1 patient, vascular malformations in 5 patients, and no abnormality in 22 patients were detected. Magnetic resonance imaging has a high success rate in detecting structural brain abnormalities, of both temporal and extratemporal locations, associated with medically intractable epilepsy syndromes. So MRI plays a primary role in planning of the treatment, primarily surgical therapy, by detecting structural epileptogenic lesions. PMID- 11870470 TI - Isotropic 3D fast FLAIR imaging of the brain in multiple sclerosis patients: initial experience. AB - The application of image registration and subtraction to detect change in multiple sclerosis (MS) disease burden on serial MR scans benefits from the use of isotropic voxels. An optimised 3D fast fluid-attenuated inversion recovery (FLAIR) sequence with 1.2- and 1.8-mm cubic voxels was compared with a 2D T2 SE sequence using standard 3-mm slices. Three-dimensional fast FLAIR and T2 SE series were obtained in 20 MS patients and 15 controls. Whole brain acquisition times for the 1.2- and 1.8-mm FLAIR were 21 and 10.5 min, respectively, for the interleaved T2 SE 16 min. Brain lesions were marked in consensus by two radiologists and the CNR was calculated in ten lesions. The mean number of lesions detected with the 1.2-mm FLAIR sequence was 115 +/- 76, compared with 85 +/- 59 for the T2 SE series ( p<0.001). The 1.8-mm FLAIR detected only 73 +/- 46 lesions. The CNR of the 1.2-mm FLAIR was significantly better than the T2 SE ( p<0.01), but not as good as the 1.8-mm FLAIR. In conclusion, isotropic 3D fast FLAIR using 1.2-mm cubic voxels is superior to the 2D T2 SE in the detection of brain lesions in MS patients. The isotropic 1.8-mm FLAIR is faster and has better contrast characteristics but lacks sensitivity. PMID- 11870471 TI - Focal status epilepticus: follow-up by perfusion- and diffusion MRI. AB - Diffusion-weighted MRI demonstrated bright right temporoparietal cortex, right hippocampus, and left cerebellum in a 63-year-old female suffering a focal convulsive status epilepticus. Hyperperfusion was noted in the right temporoparietal region. Two days later, a tendency to normalization of most of the diffusion and perfusion changes was noted, apart from the right hippocampus which became brighter on diffusion- and T2-weighted images. On the tenth day the apparent diffusion coefficient was slightly elevated, getting brighter on T2 weighted images with suspected mild post-contrast enhancement. We postulate that the discharging right hippocampus suffered cytotoxic edema, which later progressed to cell damage. PMID- 11870472 TI - Helical CT of the urinary organs. AB - Despite of the diagnostic potential of conventional CT (CCT), limitations being inherent in this technology reduce its diagnostic confidence and limit clinical CT applications as 3D imaging. Helical CT (HCT) has far overcome the limitations of CCT and has become the standard CT technology. After a short overview on the technique of HCT and its advantages over CCT, the impact of HCT on the detection of disorders of the urinary organs is discussed. Due to the high quality of 3D reconstructions, vessels are visualized free of artefacts resulting in a dramatic improvement and acceptance of CT angiography, which has become a clinically important examination in the evaluation of obstructive renal artery disease. Fast HCT provides a precise assessment of the three phases of the nephrogram and it is a prerequisite for an improved depiction of abnormal vascular perfusion and impaired tubule transit of contrast material. Helical CT enables an improved characterization of cystic mass lesions reducing the diagnosis of indeterminate masses and thus facilitating a better therapeutic management. The diagnosis of renal cell carcinomas (RCC) has improved due to an increased sensitivity in detecting small RCCs, and an increased specificity in the diagnosis of neoplastic lesions. Improved staging of RCCs is the result of accurate assessment of venous tumour extension. When planning nephron-sparing surgery 3D display of the renal tumour helps to determine the resectability of the mass depicting its relation to major renal vessels and the renal collecting system. In the evaluation of renal trauma HCT provides shorter scanning time and thus fewer artefacts in the examination of traumatized patients who cannot cooperate adequately. Three dimensional postprocessing modalities allow the assessment of the renal vascular pedicel by CT angiography and improve the demonstration of complex lacerations of the renal parenchyma. In the evaluation of the upper urinary tract unenhanced HCT has become the imaging method of choice in the diagnosis and differential diagnosis of acute flank pain since it is highly sensitive and specific in detecting calculus disease. Unenhanced HCT may furthermore demonstrate causes of flank pain unrelated to urolithiasis. Gapless volume scanning and improved resolution in the z-axis during the excretory phase enables improved visualization of the renal collecting systems and ureters, resulting in a better demonstration of intraluminal and extraluminal pathology. PMID- 11870473 TI - Multislice CT-image-guided endoscopic sinus surgery using an electromagnetic tracking system. AB - Our objective was to assess the practicability and accuracy of a computer assisted multislice CT-guided frameless electromagnetic tracking for endoscopic sinus surgery. Eighty-two patients with various paranasal sinus diseases were evaluated. Prior to surgery, axial multislice spiral-CT scans with 2.5-mm collimation, 0.8-mm reconstruction increment, and a pitch of 3 were acquired. After Ethernet transfer of the CT data set to the guidance system, coronal and sagittal images were reconstructed. For intraoperative navigation the Insta Trak System (Visualization Technology, Boston, Mass.) was used. Navigational procedures are described in detail in the paper. Accuracy was assessed by means of visual landmarks which could be clearly identified endoscopically as well as on CT images. A second parameter for accuracy was calculated by the system itself as the root mean square (RMS). The system was able to display the position of the aspirating tip relative to anatomical structures with an average accuracy of 0.70 +/- 0.40 mm. Root mean square values showed a mean value of 0.40 +/- 0.20 mm. During surgical procedures the Insta Trak System provides the surgeon with additional image-based information to the endoscopic view. The device accuracy is high and the system proves to be practicable and efficient in ENT surgery. PMID- 11870475 TI - Extranodal manifestation of Kimura's disease: ultrasound features. AB - We describe the imaging features of a 53-year-old Chinese patient with Kimura's disease (KD) who presented with a dominant right-cheek subcutaneous mass. The lesion appeared as an ill-defined mass within the subcutaneous tissues with a "wooly" echotexture on US. The differential diagnosis and a review of the literature are discussed. PMID- 11870474 TI - Percutaneous treatment of a parotid gland hydatid cyst: a possible alternative to surgery. AB - Although the most involved organs are liver and lung, hydatid cysts are very rarely seen in the head and neck region. Only a few cases with hydatid cyst in parotid gland have been reported in the literature. We present the findings of 18 months of follow-up of a case with a hydatid cyst in parotid gland treated percutaneously by using PAIR technique. To our knowledge, this is the first case of parotid gland hydatid cyst who underwent percutaneous treatment. Percutaneous treatment of parotid hydatid cyst seems to be a safe and effective procedure as a possible alternative to surgery. PMID- 11870477 TI - Idiopathic interstitial pneumonias: imaging-pathology correlation. AB - The terminology related to idiopathic interstitial pneumonia (IIP) remains confusing and in some cases wholly inaccurate. In addition, a greater understanding of the correlation between high-resolution computed tomography (HRCT) appearances and the corresponding histopathological changes found in the interstitial pneumonias has resulted in a crucial role for HRCT in the investigation of IIPs. The role of the radiologist is becoming increasingly important with a strong emphasis on establishing a diagnosis without resorting to lung biopsy. We aim to clarify the current classification of the IIPs highlighting their clinical, pathological and imaging characteristics in order to assist the radiologist in performing their increasingly important diagnostic role. PMID- 11870476 TI - Technetium-99m-sestamibi/pertechnetate subtraction scintigraphy vs ultrasonography for preoperative localization in primary hyperparathyroidism. AB - A prospective study was performed to evaluate the efficacy of technetium-99m sestamibi and technetium-99m-pertechnetate subtraction scanning and US for imaging parathyroid glands in primary hyperparathyroidism. Sixty-three patients were surgically treated for primary hyperparathyroidism (HPT). Preoperative scintigraphy and US were performed in all cases. Bilateral neck exploration was carried out on each patient. Results of radionuclide studies and US were compared with surgical and histological findings. In 57 patients with primary HPT the radionuclide scanning gave true-positive results. Four false-negative and two false-positive scintigrams were obtained. The sensitivity and the positive predictive value (PPV) of scintigraphy were 93 and 97%, respectively. Forty-one cases were correctly localized by the US. Seventeen US results were false negative and five were false positive. The sensitivity and the PPV for US were 71 and 89%, respectively. There was a statistically significant difference between the sensitivity of the scintigraphy compared with the US ( p=0.001). Sensitivities of radionuclide scans and US were higher for adenomas (100 and 83%) than for hyperplastic glands (75 and 40%). The sensitivity of technetium-99m sestamibi and technetium-99m-pertechnetate subtraction scintigraphy was significantly higher compared with US. This sensitive method could help surgeons in performing a rapid and directed parathyroidectomy. PMID- 11870478 TI - Pulmonary embolism due to cardiac hydatid disease: imaging findings of unusual complication of hydatid cyst. AB - We report two cases of primary cardiac hydatid cyst in which hydatid materials caused recurrent embolizations in pulmonary arteries and pulmonary parenchyma. Cardiac hydatid cysts may stay asymptomatic for a long time, until they reveal themselves being perforated into cardiac chambers and/or pulmonary artery or systemic circulation. The role of imaging techniques in diagnosis is discussed and the importance of dynamic enhanced CT, MR imaging, and enhanced MR angiography (MRA) is reported. Imaging findings were confirmed by surgery and pathology. Early diagnosis is essential because delayed treatment increases the morbidity and mortality rates. PMID- 11870479 TI - Radiation dose in dental radiology. AB - The aim of this study was to compare radiation exposure in panoramic radiography (PR), dental CT, and digital volume tomography (DVT). An anthropomorphic Alderson Rando phantom and two anatomical head phantoms with thermoluminescent dosimeters fixed at appropriate locations were exposed as in a dental examination. In PR and DVT, standard parameters were used while variables in CT included mA, pitch, and rotation time. Image noise was assessed in dental CT and DVT. Radiation doses to the skin and internal organs within the primary beam and resulting from scatter radiation were measured and expressed as maximum doses in mGy. For PR, DVT, and CT, these maximum doses were 0.65, 4.2, and 23 mGy. In dose-reduced CT protocols, radiation doses ranged from 10.9 to 6.1 mGy. Effective doses calculated on this basis showed values below 0.1 mSv for PR, DVT, and dose-reduced CT. Image noise was similar in DVT and low-dose CT. As radiation exposure and image noise of DVT is similar to low-dose CT, this imaging technique cannot be recommended as a general alternative to replace PR in dental radiology. PMID- 11870480 TI - Vacuum-assisted breast biopsy on digital stereotaxic table of nonpalpable lesions non-recognisable by ultrasonography. AB - The aim of this study was to evaluate accuracy of 11 G vacuum-assisted percutaneous biopsy (VAPB) carried out on digital stereotaxic table, on breast non-palpable lesions (NPLs), non-visible by US. Prospective study on 132 consecutive NPLs (126 patients) not reliably found by US; 82% showed microcalcifications. Surgical confirmation was obtained in all malignant cases and when VAPB reported atypical lesion (ductal or lobular), radial scar or atypical papillary lesion. All patients with benign results were included in a mammographic follow-up programme. Two cases could not be dealt with due to technical difficulties. One to 26 cylinders were obtained from the remaining 130 NPLs. Sixty-four lesions were surgically confirmed. Forty-six of the 47 malignancies were correctly diagnosed. In one case of a malignant tumour, an atypical lesion was classified with VAPB. All cases of histologically verified lobular carcinoma in situ, atypical ductal or lobular hyperplasia, radial scar or atypical papillary lesion were correctly diagnosed preoperatively. The remaining lesions were benign in VAPB, and after 1 year of follow-up, no false negative has been found. Based on this short-term follow-up, absolute sensitivity was 97.9%, absolute specificity 84.3% and accuracy was 99.2%. For predicting invasion, accuracy was 89.1%. Vacuum-assisted percutaneous biopsy is a very accurate technique for NPLs which are not detectable by US. It can replace approximately 90% of DSB with no important complications, avoiding scars and providing a higher level of comfort. PMID- 11870481 TI - Churg-Strauss syndrome involving the breast: a rare cause of eosinophilic mastitis. AB - Churg-Strauss syndrome is a rare immunoallergic disorder that usually affects lungs, skin and nervous system. The clinical and radiological findings of Churg Strauss disease involving the breast are reported and attention is drawn to the fact that, although uncommonly, the breast can be involved by immunological diseases. PMID- 11870482 TI - Fluid collections and juxta-articular cystic lesions of the shoulder: spectrum of MRI findings. AB - The MR imaging features of fluid collections and juxta-articular cystic lesions of the shoulder are discussed, with special focus on those related to subacromial impingement and rotator cuff tears. Other more unusual fluid collections and cystic lesions are described, including rice-bodies bursitis, idiopathic synovial osteochondromatosis, dialysis-related amyloid arthropathy, hemophilic arthropathy, infectious conditions, non-infectious inflammatory arthritis, and paralabral cysts. PMID- 11870483 TI - The intervertebral disk: a landmark for spinal diseases in children. AB - The aim of this paper is to describe the diagnostic value of the different radiological modifications of the intervertebral disk in children. Usual disk modifications include: disk-space narrowing; disk space enlargement; intervertebral disk calcifications; intervertebral disc prolapse; and signal modification in MRI. The gamuts (which are provided in the paper) of these different images must take into account the association with bone changes and the possibility of several disks' involvement. Calcifications are usually related to benign disorders. PMID- 11870484 TI - Cyclic voiding cystourethrography: is vesicoureteral reflux missed with standard voiding cystourethrography? AB - Vesicoureteral reflux (VUR) may occur intermittently and cyclic voiding cystourethrography (VCUG) can enhance the ability of the method to detect reflux. We undertook this prospective study to assess how often VUR may occur intermittently during VCUG and to evaluate the reliability of the method by performing cyclic VCUG. Two hundred seventy-five children younger than 2 years underwent two cycles of VCUG. Ninety-seven refluxing kidney-ureter units (KUU) from 68 children were identified during the two cycles. In 18 children VUR was demonstrated in the first, and in 50 children only in the second, cycle. Discrepancy between the two cycles regarding the presence and/or grade of VUR was observed in 85 KUU from 63 of 275 children (23%). In 21 of these 63 children VUR was > or = grade III. In the presence of reflux in the first cycle, discordant findings in the second cycle were found in 11 of 23 KUU (48%) or in 13 of 18 children (72.2%). In the absence of VUR in the first cycle, the second cycle disclosed reflux in 50 of 257 children (19.5%). In conclusion, intermittent VUR occurred in up to 23% of children undergoing VCUG. In more than one-third of them VUR was of major degree. Cyclic VCUG can enhance the ability of the method to detect and grade reflux. PMID- 11870485 TI - Iatrogenic intravascular pneumocephalus secondary to intravenous catheterization. AB - The presence of pneumocephalus without a history of intracranial or intrathecal procedures is a significant radiographic finding. Although pneumocephalus means a violation of the dural barrier or the presence of infection, intravascular pneumocephalus is different from intraparenchymal pneumocephalus and its benign nature must be known in the presence of intravenous catheterization. Herein, we present a case of iatrogenic intravascular pneumocephalus with CT findings. To our knowledge, there are only a few reported cases of iatrogenic intravascular pneumocephalus in the literature. Careful intravenous catheterization and diagnosis of the condition on imaging helps to prevent unnecessary treatment procedures. PMID- 11870486 TI - Guidelines and algorithms: strategies for standardization of referral criteria in diagnostic radiology. AB - Guidelines can be regarded as special forms of algorithms and have been shown to be useful tools for supporting medical decision making. With the Council Directive 97/43/Euratom recommendations concerning referral criteria for medical exposure have to be implemented into national law of all EU member states. The time- and cost-consuming efforts of developing, implementing, and updating such guidelines are balanced by the acceptance in clinical practice and eventual better health outcomes. Clearly defined objectives with special attention drawn on national and regional differences among potential users, support from organisations with expertise in evidence-based medicine, separated development of the evidence component and the recommendations component, and large-scale strategies for distribution and implementation are necessary. Editors as well as users of guidelines for referral criteria have to be aware which expectations can be met and which cannot be fulfilled with this instrument; thus, dealing with guidelines requires a new form of "diagnostic reasoning" based on medical ethics. PMID- 11870487 TI - Multidetector-row helical CT: analysis of time management and workflow. AB - The purpose of this study was to evaluate time management and workflow for multidetector-row helical CT (MDCT). Time for patient and data handling of at total of 580 patients were evaluated at two different time periods (December 1999, August 2000), each for the following baseline measurements: (a) change of clothes/instruction; (b) patient placement on the CT table/i.v. catheter; (c) CT planning and programming; (d) CT data acquisition; (e) CT data reconstruction; (f) CT data storage/printing. All imaging was performed on a Somatom Volume Zoom (Siemens, Erlangen, Germany). Time measurements summarized for different CT protocols revealed the following: (a) 5:01 min (+/- 2.06 min); (b) 4:36 min (+/- 2.43 min); (c) 4:11 min (+/- 2.55 min); (d) 0:43 min (+/- 0.15 min); (e) 6:59 min (+/- 2.39 min); (f) 09:51 min (+/- 3.51 min). Planning and programming was most time-consuming for CT angiography, whereas chest and abdominal CT needed only 3:26 and 3:30 min, respectively. Reconstruction time was highest for HRCT (9:22 min) and CTA (9:03 min). Data storage/printing was most time-consuming for HRCT (13:02 min), followed by combined neck-chest-abdomen examinations (12:19 min). Comparing the two time periods, during which a software update was performed, a mean time reduction of 4:31 min per patient (15%, p<0.001) was achieved. Whereas CT data acquisition time is no longer a problem with MDCT, patient management, data reconstruction, and data storage are the most time-consuming parts. Well trained technicians, state-of-the-art workstations, and fast networking are the most important factors to improve workflow. PMID- 11870490 TI - Repositioning of malpositioned or flipped central venous catheters. AB - Primary misplaced or secondary flipped implanted catheters are located mostly in the right jugular vein. We demonstrate an effective method to replace fix implanted catheters such as Ports, Grochomg or Hickman catheters. Using a femoral venous approach, replacement into the superior vena cava can easily be done with a Sidewinder 1 catheter which is hooked over the misplaced central venous approach. In all our patients the method was successful. The repositioning technique described is simple, fast and has low costs. We can keep sterile conditions and do not need to solve the catheters' fixation. PMID- 11870488 TI - Comparison in myelography between iodixanol 270 and 320 mgI/ml and iotrolan 300 mgI/ml: a multicentre, randomised, parallel-group, double-blind, phase III trial. AB - The objective of the trial was to compare the safety and efficacy of the non ionic, dimeric, isotonic contrast medium iodixanol (Visipaque 270 and 320 mgI/ml) with those of iotrolan (Isovist 300 mgI/ml) in myelography. After lumbar or cervical puncture, 315 patients were examined in a multicentre, double-blind, randomised, comparative myelography study. Image quality, changes in vital signs, immediate and delayed adverse events were registered. There was a tendency for better images with iodixanol 320 than with iodixanol 270 and iotrolan 300, but the overall quality was good or excellent with all products. The frequency of patients reporting adverse events and headache varied much across centres, but there was no statistically significant difference between the contrast media. The incidence of events was higher after lumbar puncture than after cervical puncture, in women rather than in men, and after puncture with a 22-gauge (G) bevel-tipped needle compared with a 24 G Sprotte needle. The frequency of headache did not correlate with the absence of pathology. The higher iodine concentration in iodixanol 320 could be an advantage for film quality. When compared with iotrolan 300, iodixanol 320 and 270 give similar incidences of adverse events, including headache. PMID- 11870491 TI - Caroli's disease: central dot sign re-examined by CT arteriography and CT during arterial portography. PMID- 11870489 TI - Gadomer-17-enhanced 3D navigator-echo MR angiography of the pulmonary arteries in pigs. AB - The goal of this study was visualisation and quality assessment of the pulmonary arteries in pigs with modified navigator-echo magnetic resonance angiography using an intravascular contrast agent. Five sedated pigs were examined in a clinical 1.5-T system with modified three-dimensional navigator-echo magnetic resonance angiography (slice thickness 3 mm, pixel size 2.4x1.8 mm2) to evaluate the pulmonary arteries. Using a phased-array four-element thorax coil the entire thorax was scanned before and after intravenous infusion of a gadolinium-based intravascular contrast agent. Assessment of image quality, enhancement-related contrast-to-noise ratio (CNR) measurements and improvement of visibility of peripheral pulmonary vessels was performed. Improvement of quality using Gadomer 17 was found for smaller vessels; pulmonary trunks and the main pulmonary arteries were sufficiently imaged without enhancement. Mean rise of CNR measured in the pulmonary trunks was 28.64% ( P=0.0002), mean rise of CNR in the main pulmonary arteries and the segmental arteries were 79.6% and 148.2%, respectively. Mean distance between the visible peripheral end of 60 sub segmental arteries and the inner thoracic wall was 12.2 +/- 0.4 mm, and was significantly ( P=0.00002) reduced after contrast infusion to 8.0 +/- 0.4 mm. The combination of inherent flow sensitivity of navigator-echo angiography and Gadomer-17 proved effective for imaging of the pulmonary arteries. In contrast to standard contrast-enhanced pulmonary MRA studies, breath holding is not required. Further studies and the evaluation of findings of patients suffering from pulmonary embolism are needed to evaluate the possible benefits of a higher spatial resolution which is achievable using navigator-echo techniques in contrast to the higher temporal resolution of ultra-fast pulmonary MRA. PMID- 11870498 TI - March 2002. PMID- 11870493 TI - ESUR Newsletter. PMID- 11870492 TI - Lipoma of the internal auditory canal: MR findings. PMID- 11870499 TI - Cimetidine in colorectal cancer--are the effects immunological or adhesion mediated? PMID- 11870500 TI - Cimetidine increases survival of colorectal cancer patients with high levels of sialyl Lewis-X and sialyl Lewis-A epitope expression on tumour cells. AB - Cimetidine has been shown to have beneficial effects in colorectal cancer patients. In this study, a total of 64 colorectal cancer patients who received curative operation were examined for the effects of cimetidine treatment on survival and recurrence. The cimetidine group was given 800 mg day(-1) of cimetidine orally together with 200 mg day(-1) of 5-fluorouracil, while the control group received 5-fluorouracil alone. The treatment was initiated 2 weeks after the operation and terminated after 1 year. Robust beneficial effects of cimetidine were noted: the 10-year survival rate of the cimetidine group was 84.6% whereas that of control group was 49.8% (P<0.0001). According to our previous observations that cimetidine blocked the expression of E-selectin on vascular endothelium and inhibited the adhesion of cancer cells to the endothelium, we have further stratified the patients according to the expression levels of sialyl Lewis antigens X (sL(x)) and A (sL(a)). We found that cimetidine treatment was particularly effective in patients whose tumour had higher sL(x) and sL(a) antigen levels. For example, the 10-year cumulative survival rate of the cimetidine group with higher CSLEX staining, recognizing sL(x), of tumours was 95.5%, whereas that of control group was 35.1% (P=0.0001). In contrast, in the group of patients with no or low levels CSLEX staining, cimetidine did not show significant beneficial effect (the 10-year survival rate of the cimetidine group was 70.0% and that of control group was 85.7% (P=n.s.)). These results clearly indicate that cimetidine treatment dramatically improved survival in colorectal cancer patients with tumour cells expressing high levels of sL(x) and sL(a). PMID- 11870501 TI - Recovery of zeta-chain expression and changes in spontaneous IL-10 production after PSA-based vaccines in patients with prostate cancer. AB - Circulating T lymphocytes of patients with prostate cancer have been reported to have functional deficits, including low or absent zeta-chain expression. To determine whether these functional impairments could be reversed by prostate specific antigen-based vaccination therapy, 10 patients treated with recombinant human prostate specific antigen plus GM-CSF and eight others receiving prostate specific antigen plus oil emulsion in two pilot clinical trials were evaluated prior to and after vaccination for several immunologic end points, including zeta chain expression and cytokine production by circulating T cells as well as the frequency of T cells able to respond to prostate specific antigen in ELISPOT assays. The flow cytometry assay for zeta-chain expression was standardized to allow for a reliable comparison of pre- vs post-vaccination samples. Prior to therapy, the patients were found to have significantly lower zeta-chain expression in circulating CD3(+) cells and a higher percentage of zeta-chain negative CD3(+) and CD4(+) cells than normal donors. The patients' peripheral blood mononuclear cells spontaneously produced more IL-10 ex vivo than those of normal controls. After vaccination, recovery of zeta-chain expression was observed in 50% of patients in both clinical trials. Also, spontaneous IL-10 secretion by peripheral blood mononuclear cells decreased following immunotherapy in patients treated with prostate specific antigen and GM-CSF. The frequency of prostate specific antigen-reactive T cells was detectable in 7 out of 18 patients vs 4 out of 18 patients prior to vaccination. Only one of 18 patients was a clinical responder. The vaccine had stimulatory effects on the patients' immune system, but post-vaccine immune recovery could not be correlated to progression free survival in this small cohort of patients with prostate cancer. PMID- 11870502 TI - Combination chemotherapy with or without s.c. IL-2 and IFN-alpha: results of a prospectively randomized trial of the Cooperative Advanced Malignant Melanoma Chemoimmunotherapy Group (ACIMM). AB - The purpose of this randomized trial was to evaluate the efficacy of combination chemoimmunotherapy compared with chemotherapy alone. A total of 124 patients were randomized to receive intravenous cisplatin (35 mg m(-2), days 1-3), carmustine (150 mg m(-2), day 1, cycles 1 and 3 only), dacarbacine (220 mg m(-2), days 1-3) and oral tamoxifen (20 mg m(-2), daily) in combination with (n=64) or without (n=60) sequential subcutaneous IL-2 and IFN-alpha. In those patients who received sequential immunotherapy, each cycle of chemotherapy was followed by outpatient s.c. IL-2 (10 x 10(6) IU m(-2), days 3-5, week 4; 5 x 10(6) IU m(-2), days 1, 3, 5, week 5) and s.c. IFN-alpha (5 x 10(6) IU m(-2), day 1, week 4; days 1, 3, 5, week 5). The overall response rate of patients treated with the combination of chemotherapy and IL-2/IFN-alpha was 34.3% with seven complete responses (10.9%) and 15 partial responses (23.4%). In patients treated with chemotherapy, only, the overall response rate was 29.9% with eight complete responses (13.3%) and 10 partial responses (16.6%). There was no significant difference in median progression free survival (0 months vs 4 months) and in median overall survival (12 months vs 13 months) for combined chemoimmunotherapy and for chemotherapy, respectively. PMID- 11870503 TI - The free beta-subunit of human chorionic gonadotropin as a prognostic factor in renal cell carcinoma. AB - The free beta-subunit of human chorionic gonadotropin beta is expressed in several nontrophoblastic tumours and this is usually associated with aggressive disease. Little is known about human chorionic gonadotropin beta expression in renal cancer. We determined the pretreatment levels of human chorionic gonadotropin beta in serum of patients with renal cell carcinoma, and studied whether elevated levels predicted the clinical outcome. Serum samples were collected before surgery from 177 patients with renal cell carcinoma and from 84 apparently healthy controls. Human chorionic gonadotropin beta in serum was measured by a highly sensitive time-resolved immunofluorometric assay. The prognostic value of human chorionic gonadotropin beta, and of usual clinical and pathological variables was analyzed by the Kaplan-Meier method, the log rank test and Cox multiple hazard regression. The serum concentrations of human chorionic gonadotropin beta were increased in 23% of the renal cell carcinoma patients and they were significantly higher in patients with renal cell carcinoma than in controls (P<0.0001). The concentrations did not correlate with clinical stage and histopathological grade, but patients with increased human chorionic gonadotropin beta levels had significantly shorter survival time than those with levels below the median (cut-off 1.2 pmol l(-1), P=0.0029). In multivariate analysis human chorionic gonadotropin beta, tumour stage and grade were independent prognostic variables. The serum concentration of human chorionic gonadotropin beta is an independent prognostic variable in renal cell carcinoma. The preoperative value of human chorionic gonadotropin beta in serum may be used to identify patents with increased risk of progressive disease. PMID- 11870504 TI - Gemcitabine and cisplatin in a multimodality treatment for locally advanced non small cell lung cancer. AB - The role of new cytotoxic agents like gemcitabine has not yet been proven in the neoadjuvant settings. We designed a phase II study to test the feasibility of using gemcitabine and cisplatin before local treatment for stage III non-small cell lung cancer patients. Patients received three cycles of induction chemotherapy of gemcitabine (1000 mg m(-2), days 1, 8, 15) and cisplatin (90 mg m(-2), day 15) every 4 weeks before evaluation for operability. Operable patients underwent radical resection. Inoperable patients and patients who had incomplete resection received concurrent chemoradiotherapy with daily low dose cisplatin. All patients who did not progress after local treatment received three more cycles of adjuvant chemotherapy of gemcitabine and cisplatin. Fifty-two patients received induction treatment. Two patients had complete response and 31 patients had partial response (response rate 63.5%) after induction chemotherapy. Thirty six patients (69%) were operable. Eighteen patients (35%) had their tumours completely resected. Two patients had pathological complete response. Median overall survival was 19.1 months, projected 1-year survival was 66% and 2-year survival was 34%. Three cycles of gemcitabine and cisplatin is effective and can be used as induction treatment before surgery for locally advanced non-small cell lung cancer patients. PMID- 11870505 TI - Monocomponent chemoembolization in oral and oropharyngeal cancer using an aqueous crystal suspension of cisplatin. AB - Intensification of intra-arterial chemotherapy with high-dose cisplatin and concomitant reduction of toxicity under the conditions of the head and neck was aimed at by combination of antineoplastic activity and embolizing effect in the same pharmacon. A cisplatin suspension in normal saline (5 mg in 1 ml) with precipitation of microembolizing cisplatin crystals was prepared. No additional pharmacons. Cisplatin dosage was 150 mg m(-2), maximum absolute dose 300 mg, maximum amount of fluid 60 ml. Thirty patients (UICC-Classification of tumours: I/2 patients, II/6, III/2; IV/20) were treated in a neoadjuvant setting with superselective chemoembolization using the cisplatin suspension. A control group (n=30) with the same tumour and nodal staging was treated with a usual cisplatin solution (150 mg m(-2) dissolved in 500 ml saline). In both groups, parallel intravenous infusion of sodium thiosulphate (9 g m(-2)). Endpoints were toxicity and response. Continuation of treatment by surgery or radiation. Overall remission was 70% in the study group and 46.7% in the control group after one cycle respectively. Systemic side-effects were very low (grade I WHO) in both groups. Side-effects were found to be similar to post-embolization syndrome (swelling, mild to moderate pain, leucocytosis without fever) in the study group. Chemoembolization in the head and neck area can be carried out routinely using this method. PMID- 11870506 TI - Measurement of serum total and free prostate-specific antigen in women with colorectal carcinoma. AB - We investigated the diagnostic value and the relationship with clinicopathological features of total and free prostate-specific antigen by measuring the concentrations of these markers in the sera of 75 women with colorectal carcinoma and in 30 healthy women. Measurements were performed by immunoradiometric assay which utilizes monoclonal and polyclonal anti-prostate specific antigen antibodies; the lowest detection level for both markers was 0.01 ng ml(-1). Free prostate-specific antigen levels were significantly higher in women with colorectal carcinoma than healthy women (P=0.006). The percentage of free prostate-specific antigen predominant (free prostate-specific antigen/total prostate-specific antigen >50%) subjects was 20% in colorectal carcinoma patients and 3.3% in healthy women (P=0.035). Cut-off values were 0.34 ng ml(-1) for total prostate-specific antigen and 0.01 ng ml(-1) for free prostate-specific antigen. In women with colorectal carcinoma, total prostate-specific antigen positivity was 20% and free prostate-specific antigen positivity was 34.6%. When compared to negatives, total prostate-specific antigen positive patients had a lower percentage of well-differentiated (P=0.056) and early stage (stages I and II) tumours (P=0.070). However, patients with predominant free prostate-specific antigen, had a higher percentage of well-differentiated (P=0.014) and early stage tumours (P=0.090) than patients with predominant bound prostate-specific antigen. In conclusion, although the sensitivity of free prostate-specific antigen predominancy is low (20%), in distinguishing women with colorectal carcinoma than healthy women, its specificity is high (96.7%). Free prostate-specific antigen predominancy tends to be present in less aggressive tumours. These findings may indicate clinical significance of preoperative measurement of serum total and free prostate-specific antigen in women with colorectal carcinoma. PMID- 11870507 TI - Pretargeted adjuvant radioimmunotherapy with yttrium-90-biotin in malignant glioma patients: a pilot study. AB - In a previous study we applied a three-step avidin-biotin pretargeting approach to target 90Y-biotin to the tumour in patients with recurrent high grade glioma. The encouraging results obtained in this phase I-II study prompted us to apply the same approach in an adjuvant setting, to evaluate (i) time to relapse and (ii) overall survival. We enrolled 37 high grade glioma patients, 17 with grade III glioma and 20 with glioblastoma, in a controlled open non-randomized study. All patients received surgery and radiotherapy and were disease-free by neuroradiological examinations. Nineteen patients (treated) received adjuvant treatment with radioimmunotherapy. In the treated glioblastoma patients, median disease-free interval was 28 months (range=9-59); median survival was 33.5 months and one patient is still without evidence of disease. All 12 control glioblastoma patients died after a median survival from diagnosis of 8 months. In the treated grade III glioma patients median disease-free interval was 56 months (range=15 60) and survival cannot be calculated as only two, within this group, died. Three step radioimmunotherapy promises to have an important role as adjuvant treatment in high grade gliomas, particularly in glioblastoma where it impedes progression, prolonging time to relapse and overall survival. A further randomized trial is justified. PMID- 11870509 TI - Estimates of the likely prophylactic effect of tamoxifen in women with high risk BRCA1 and BRCA2 mutations. AB - The development of breast cancer control strategies in women at high genetic risk of breast cancer is an important issue. The likely benefit of chemopreventive approaches is of particular interest. Tamoxifen tends to be more effective in both prevention and treatment of oestrogen receptor positive tumours than oestrogen receptor negative. In this study, we combine the oestrogen-receptor specific effects of tamoxifen from randomized preventive or therapeutic trials with the oestrogen receptor status of tumours in BRCA1 and BRCA2 mutation positive women from published tumour surveys to obtain estimates of the likely effect of tamoxifen administration in mutation carriers. We used a simple two stage procedure to estimate the benefit as a weighted average of the effect on oestrogen receptor positive tumours and oestrogen receptor negative, and using a more complex hierarchical modelling approach. Using the simple procedure and deriving the estimates of benefit from both primary prevention and therapeutic trials, we obtain an estimated reduction in risk of breast cancer from administration of tamoxifen in BRCA1 mutation positive women of 13% (RR=0.87, 95% CI 0.68--1.11). The corresponding estimated reduction in BRCA2 mutation positive women was 27% (RR=0.73, 95% CI 0.59--0.90). Using the more complex models gave essentially the same results. Using only the primary prevention trials gave smaller estimates of benefit in BRCA1 carriers but larger estimates in BRCA2, in both cases with wider confidence intervals. The benefit of prophylactic use of tamoxifen in BRCA1 mutation carriers is likely to be modest, and the effect in BRCA2 mutation carriers somewhat greater. PMID- 11870508 TI - Infusional 5-fluorouracil, cisplatin and mitomycin C in advanced gastric cancer: a low cost effective regimen. AB - Recently, we reported a highly active regimen in advanced gastric cancer including a weekly administration of cisplatin, epidoxorubicin, leucovorin, 5 fluorouracil with the support of filgrastim. In order to simplify the administration and to decrease the toxicity of these drugs, mainly epidoxorubicin induced alopecia, we designed a regimen including an infusional 5-fluorouracil schedule according to the de Gramont regimen, cisplatin and mitomycin C replacing epidoxorubicin. Forty-five patients with advanced or metastatic gastric cancer were treated with cisplatin 50 mg m(-2) i.v. on day 1, every 2 weeks, 6S stereoisomer-leucovorin 100 mg m(-2) i.v. followed by 5-fluorouracil 400 mg m(-2) i.v. bolus and 600 mg m(-2) i.v. in a 22-h infusion, on days 1 and 2, every 2 weeks, and mitomycin C 7 mg m(-2) i.v. bolus on day 2, every 6 weeks. Grades 3-4 toxicities (National Cancer Institute-Common Toxicity Criteria) consisted mainly of neutropenia and thrombocytopenia. Five patients had a complete response and 16 had a partial response for an overall response rate of 46.7% (95% confidence interval, 32.1-61.2%). The median survival was 11 months. The combination of cisplatin, 5-fluorouracil and leucovorin according to de Gramont, and mitomycin C seems to be an active and safe regimen in the treatment of advanced gastric cancer. Because of its low cost it may be suggested for patients not enrolled into clinical trials. PMID- 11870510 TI - Prognostic significance of dihydropyrimidine dehydrogenase expression in breast cancer. AB - We have investigated dihydropyrimidine dehydrogenase expression as a prognostic marker in breast cancer. A total of 119 women with breast cancer undergoing surgery between 1985 and 1996 were included in this study. Eighty-seven patients were treated with postoperative chemotherapy including 5-fluorouracil or 5 fluorouracil derivatives. Fifty-nine (50%) of 119 patients were determined to be immunostaining-positive for dihydropyrimidine dehydrogenase. There was no significant difference between dihydropyrimidine dehydrogenase staining and tumour size, lymph node status, clinical stage, oestrogen receptor status, histologic grade, or 5-fluorouracil administration. When evaluated in patients treated with 5-fluorouracil or 5-fluorouracil derivatives, patients with dihydropyrimidine dehydrogenase-positive tumours had a significantly (P<0.05) poorer disease-free survival compared to those with dihydropyrimidine dehydrogenase-negative tumour. No conclusion can be drawn about the prognostic impact of dihydropyrimidine dehydrogenase status in patients who were not treated with 5-fluorouracil regimes due to the small number of such cases in this series. Lymph node and dihydropyrimidine dehydrogenase status were independent prognostic factors for disease-free survival, and lymph node status for overall survival using multivariate analysis. In conclusion, dihydropyrimidine dehydrogenase is a possible prognostic factor in patients with breast cancer treated with 5 fluorouracil or 5-fluorouracil derivatives. PMID- 11870511 TI - Clinical and economic impact of multiple gated acquisition scan monitoring during anthracycline therapy. AB - The clinical and economic impacts of monitoring cardiac function in patients given doxorubicin have yet to be determined, especially in relation to patient age, cumulative doxorubicin dose, and the relative efficacies of doxorubicin based vs alternative regimens. We developed a decision analysis model that includes these factors to estimate the incremental survival benefit and cost effectiveness of using multiple gated acquisition scans to measure left ventricular ejection fraction before and during doxorubicin chemotherapy. Probability distributions for the incidences of abnormal left-ventricular ejection fraction findings and congestive heart failure were derived from a retrospective review of 227 consecutive cases at The University of Michigan Medical Center and published findings. Multiple gated acquisition-scan monitoring minimally improved the probability of 5-year survival (<1.5% in the base--case scenario). For patients who received up to 350 mg m(-2) of doxorubicin, multiple gated acquisition-scan screening had an incremental cost of $425 402 per life saved for patients between the ages of 15--39. This incremental cost markedly decreased to $138 191, for patients between the ages of 40--59, and to $86 829 for patients older than 60 years. The small gain in 5-year survival probability secondary to multiple gated acquisition scan monitoring doubled for all age groups when the average cumulative dose for doxorubicin reached 500 mg m(-2). Variations in the cure rate differences between the doxorubicin and alternative regimens had insignificant effects on the improvement in 5-year survival rates from multiple gated acquisition-scan screening. The use of multiple gated acquisition scans for pretreatment screening appears to be more cost-effective for patients who are 40 years or older, when cumulative doxorubicin dose is 350 mg m(-2) or less. PMID- 11870512 TI - A randomized trial of specialist genetic assessment: psychological impact on women at different levels of familial breast cancer risk. AB - The aim was to compare the psychological impact of a multidisciplinary specialist genetics service with surgical provision in women at high risk and those at lower risk of familial breast cancer. Women (n=735) were randomized to a surgical consultation with (trial group) or without (control group) specialist genetic risk assessment and the possible offer of presymptomatic genetic testing. Participants completed questionnaires before and immediately after the consultation to assess anxiety, cancer worry, perceived risk, interest in genetic testing and satisfaction. Responses of subgroups of women stratified by clinicians as low, moderate, or high risk were analyzed. There were no significant main effects of study intervention on any outcome variable. Regardless of risk information, there was a statistically significant reduction in state anxiety (P<0.001). Reductions in cancer worry and perceived risk were significant for women at low or moderate risk (P<0.001) but not those at high risk, and satisfaction was significantly lower in the high risk group (P<0.001). In high risk women who received specialist genetic input, there was a marginally significant trend towards increased perceived risk. The effect of risk information on interest in genetic testing was not significant. Breast care specialists other than geneticists might provide assessments of breast cancer risk, reassuring women at reduced risk and targeting those at high risk for specialist genetic counselling and testing services. These findings are discussed in relation to the existing UK Calman-Hine model of service delivery in cancer genetics. DOI: 10.1038/sj/bjc/6600051 www.bjcancer.comCopyright 2002 The Cancer Research Campaign PMID- 11870513 TI - Screening for oesophageal neoplasia in patients with head and neck cancer. AB - Due to advanced disease at the time of diagnosis the prognosis of oesophageal cancer is generally poor. As mass screening for oesophageal cancer is neither feasible nor reasonable, high-risk groups should be identified and surveilled. The aim of this study was to define the risk of oesophageal cancer in patients with (previous) head and neck cancer. A total of 148 patients with (previous) head and neck cancer were prospectively screened for oesophageal cancer by video oesophagoscopy and random oesophageal biopsies. Even in a macroscopically normal looking oesophagus, four biopsy specimens were taken every 3 cm throughout the entire length of the squamous oesophagus. Low- or high-grade squamous cell dysplasia was detected histologically in 10 of the 148 patients (6.8%). All but one dysplasias were diagnosed synchronously with the head and neck cancers. In addition, oesophageal squamous cell carcinoma was diagnosed in 11 of the 148 patients (7.4%). Most invasive cancers (63.6%) occurred metachronously. The risk of squamous cell neoplasia of the oesophagus is high in patients with (previous) head and neck cancer. Surveillance is recommended in this high-risk group. PMID- 11870514 TI - Vascular phenotype in angiogenic and non-angiogenic lung non-small cell carcinomas. AB - We have previously described a group of non-small cell lung carcinomas without morphological evidence of neo-angiogenesis. In these tumours neoplastic cells fill up the alveoli and the only vessels present appear to belong to the trapped alveolar septa. In the present study we have characterised the phenotype of the vessels present in these non-angiogenic tumours, in normal lung and in angiogenic non-small cell lung carcinomas. The vessels, identified by the expression of CD31, were scored as mature when expressing the epitope LH39 in the basal membrane and as newly formed when expressing alphaVbeta3 on the endothelial cells and/or lacking LH39 expression. In the nine putative non-angiogenic cases examined, the vascular phenotype of all the vessels was the same as that of alveolar vessels in normal lung: LH39 positive and alphaVbeta3 variable or negative. Instead in 104 angiogenic tumours examined, only a minority of vessels (mean 13.1%; range 0--60%) expressed LH39, while alphaVbeta3 (in 45 cases) was strongly expressed on many vessels (mean 55.5%; range 5--90%). We conclude that in putative non-angiogenic tumours the vascular phenotype is that of normal vessels and there is no neo-angiogenesis. This type of cancer may be resistant to some anti-angiogenic therapy and different strategies need to be developed. PMID- 11870515 TI - Expression of oestrogen receptor beta (ERbeta1) protein in human breast cancer biopsies. AB - Oestrogen action is mediated via specific receptors that act as ligand-activated transcription factors. A monoclonal antibody specific to the C-terminus of human oestrogen receptor beta has been characterized and the prevalence of expression of oestrogen receptor beta protein investigated in a well defined set of breast cancers. Reverse transcription-polymerase chain reaction analysis of RNA from tissue biopsies detected oestrogen receptor beta in all samples examined. The anti-oestrogen receptor beta antibody cross reacted specifically with both long (approximately 59 Kd) and short (approximately 53 Kd) forms of recombinant oestrogen receptor beta. Western blot analysis of breast tumours contained both forms of oestrogen receptor beta protein although in some samples lower molecular weight species (32--45 Kd) were identified. Fifty-one breast cancer biopsies were examined using immunohistochemistry; 41 (80%) were immunopositive for oestrogen receptor alpha, 48 (94%) were immunopositive for oestrogen receptor beta and 38 (74.5%) co-expressed both receptors. Expression of oestrogen receptor beta was exclusively nuclear and occurred in multiple cell types. There was no quantitative relationship between staining for the two ERs although in tumours in which both receptors were present immunoexpression of oestrogen receptor alpha was invariably more intense. The significance of oestrogen receptor beta protein expression in breast cancers to therapy remains to be determined but the availability of a well characterized antibody capable of detecting oestrogen receptor beta in archive material will facilitate the process. PMID- 11870516 TI - Reduced PTEN expression in the pancreas overexpressing transforming growth factor beta 1. AB - PTEN is a candidate tumour suppressor gene and frequently mutated in multiple cancers, however, not in pancreatic cancer. Recently, it has been demonstrated that PTEN expression is regulated by TGF-beta1. Using TGF-beta1 transgenic mice (n=7) and wildtype littermates (n=6), as well as pancreatic tissues obtained from organ donors (n=10) and patients with pancreatic cancer (n=10), we assessed the expression of PTEN by means of immunohistochemistry and semiquantitative PCR analysis. In addition, PANC-1 cells were treated with TGF-beta1 in vitro and the levels of PTEN mRNA were determined in these cells. In human pancreatic cancers PTEN mRNA levels were significantly decreased (P<0.05). In addition, in the pancreas of TGF-beta1 transgenic mice the expression of PTEN was significantly reduced (P<0.01), as compared to wildtype littermates and incubation of PANC-1 cells with TGF-beta1 decreased PTEN mRNA levels after 24 h. Inasmuch as TGF-beta1 decreases PTEN expression in human pancreatic cancer cells and human pancreatic cancers overexpress TGF-beta1, the reduced expression of PTEN in pancreatic cancer may be mediated by TGF-beta1 overexpression. Thus, although PTEN is not mutated in pancreatic cancers, the reduction of its expression may give pancreatic cancer cells an additional growth advantage. PMID- 11870517 TI - E7 proteins from oncogenic human papillomavirus types transactivate p73: role in cervical intraepithelial neoplasia. AB - In common with other E2F1 responsive genes such as p14(ARF) and B-myb, the promoter of p73 is shown to be positively regulated in cell lines and primary human keratinocytes by E7 proteins from oncogenic human papillomavirus (HPV) types 16, 18, 31 and 33, but not HPV 6. Mutational analysis revealed that transactivation of the p73 promoter by HPV 16E7 requires association with pRb. Expression of p73 in normal cervical epithelium is confined to the basal and supra-basal layers. In contrast, expression in neoplastic lesions is detected throughout the epithelium and increases with grade of neoplasia, being maximal in squamous cell cancers (SCC). Deregulation of expression of the N-terminal splice variant p73Delta2 was observed in a significant proportion of cancers, but not in normal epithelium. The frequent over-expression of p73Delta2, which has recognized transdominant properties, in malignant and pre-malignant lesions suggests a role in the oncogenic process in cervical epithelium. PMID- 11870518 TI - Variants of the long control region and the E6 oncogene in European human papillomavirus type 16 isolates: implications for cervical disease. AB - High-risk human papillomavirus types, especially type 16, are risk factors for cervical cancer. Preliminary studies suggest that HPV16 polymorphisms in the long control region or in the E6 gene may alter the oncogenic potential of the virus. This could partially explain why some lesions progress to cancer while others do not. A systematic study combining the long control region and E6 has not been undertaken. This prompted us to investigate the long control region and the E6 in northern European women infected with human papillomavirus 16. We identified the sequence variations of both regions and investigated the long control region promoter activity among various isolates. In addition, we correlated the distribution of long control region and E6 polymorphisms with disease status. We analyzed 45 samples from Swedish and Finnish women. The long control region and the E6 gene were sequenced after polymerase chain reaction long control region fragments of six European isolates covering the majority of polymorphisms in this region were ligated into the pALuc vector and used for luciferase assays. In European HPV16 isolates, polymorphisms in the long control region are more frequent than in the E6 gene. Nevertheless, the promoter function was slightly increased in only one of the tested European long control region variants. In addition, we found a specific European E6 variant, L83V, to be enriched in high grade lesions and cancer rather than a specific European long control region variant. The difference in oncogenicity between European HPV16 genotypes is more probably due to an altered property of the corresponding E6 proteins rather than to an altered activity of the P97 promoter. PMID- 11870519 TI - Identification of molecular markers for the early detection of human squamous cell carcinoma of the uterine cervix. AB - To identify novel cellular genes that could potentially act as predictive molecular markers for human cervical cancer, we employed RT--PCR differential display, reverse Northern and Northern blot analysis to compare the gene expression profiles between squamous cell carcinoma biopsies and adjacent histo pathological normal epithelium tissues. Twenty-eight cDNA clones were isolated that were demonstrated to be consistently over-expressed in squamous cell cervical cancer biopsies of FIGO stages 1B to 3B. Most importantly, it was observed that, in addition to their over-expression in cancer lesions, some of these genes are upregulated in the presumably histo-pathological normal adjacent tissues. Of particular interest is clone G30CC that has been identified to be the gene that encodes S12 ribosomal protein. When employed for RNA--RNA in situ hybridization experiments, expression of G30CC could be detected in the immature basal epithelial cells of histo-pathological normal tissues collected from cervical cancer patients of early FIGO stages. In comparison, the expression of G30CC was not detected in cervical tissues collected from patients admitted for surgery of non-malignant conditions. These results allow the distinct possibility of employing the ribosomal protein S12 gene as an early molecular diagnostic identifier for the screening of human cervical cancer and a potential target employed for cancer gene therapy trials. PMID- 11870520 TI - Methylation status of oestrogen receptor-alpha gene promoter sequences in human ovarian epithelial cell lines. AB - We have determined the methylation status of the CpG island of the oestrogen receptor alpha gene in seven human ovarian cell lines. Cell lines expressing oestrogen receptor alpha showed no evidence of hypermethylation. In three of four cell lines that produced no detectable oestrogen receptor alpha protein, hypermethylation was observed at the NotI site of the CpG island. These results indicate that aberrant hypermethylation may be responsible for a significant proportion of epithelial ovarian tumours in which oestrogen receptor alpha expression is lost. PMID- 11870521 TI - IL-4 receptors on human medulloblastoma tumours serve as a sensitive target for a circular permuted IL-4-Pseudomonas exotoxin fusion protein. AB - Cytotoxins directed to interleukin-4 receptors have shown to mediate relatively selective cytotoxicity against a variety of human cancer cells in vitro and in vivo. In an ongoing Phase I clinical trial, a recombinant protein comprised of circularly permuted IL-4 fused to a mutated form of Pseudomonas exotoxin (the fusion protein termed IL-4(38-37)-PE38KDEL or cpIL4-PE) has shown antitumour activity against malignant glioma. Human medulloblastomas are neuroectodermal tumours that occur in children and have a poor prognosis. The goal of this study was to determine whether human medulloblastoma derived cell lines express interleukin-4 receptor and whether interleukin-4 receptor expression is accompanied by sensitivity to cpIL4-PE. Medulloblastoma cell lines express interleukin-4 receptor at the protein and mRNA levels as determined by binding, indirect immunofluorescence and RT--PCR studies. These cells expressed IL-4Ralpha (also known as IL-4Rbeta) and IL-13Ralpha1 (also known as IL-13Ralpha') chains, however common gamma(c), a component of the interleukin-4 receptor system in immune cells was not detected. Consistent with the expression of IL-4R, cpIL4-PE was found to be highly and specifically cytotoxic to four of five medulloblastoma cell lines. Susceptibility of medulloblastoma cell lines to cpIL4-PE seemed to correlate closely to the functional IL-4 binding sites in general as demonstrated by 125I-IL-4 binding, but did not seem to correlate with mRNA or cell surface immunoreactive receptor protein expression. The sensitivity of medulloblastoma cells to cpIL4-PE could be eliminated by concurrent incubation with IL-4 or IL 13, but not with IL-2. None of these cell lines showed any change in proliferation upon treatment with exogenous IL-4. These studies establish the interleukin-4 receptor as a medulloblastoma-associated target for possible tumour directed cancer therapy. Further studies are warranted to investigate interleukin 4 receptor expression in primary medulloblastoma tumours and sensitivity to cpIL 4PE in vitro and in vivo. PMID- 11870522 TI - Recurrent growth factor starvation promotes drug resistance in human leukaemic cells. AB - Multi-drug resistance can be induced by various environmental stresses including an exposure to chemical drugs and X-ray irradiation. In addition, hypo-nutritive conditions are known to promote multi-drug resistance in solid tumours. To understand the importance of nutritive conditions in the development of drug resistance in non-solid tumours and to know whether a transient malnutrition could induce a permanent reduction in drug sensitivity, leukaemic cells were transiently cultured under growth factor-starved conditions. Granulocyte macrophage colony-stimulating factor-dependent human leukaemic MO7e cells were cultured in the absence of granulocyte-macrophage colon-stimulating factor for 2 weeks, during which the majority of the cells died, and the minor viable cells were expanded in the presence of granulocyte-macrophage colon-stimulating factor for following 1 week. This procedure was repeated three times, and the surviving cells were cloned by limiting dilution. These clones underwent G1 arrest in the absence of granulocyte-macrophage colon-stimulating factor, while parental cells underwent apoptosis. Interestingly, activities of the downstream targets of granulocyte-macrophage colon-stimulating factor receptor were regulated in a granulocyte-macrophage colon-stimulating factor-independent manner, indicating that the ligand-independent activation of granulocyte-macrophage colon stimulating factor receptor had not taken place. Moreover, the 4--7-fold increases in IC(50) for etoposide and the 2--6-fold increase in IC(90) for doxorubicin was observed. Furthermore, Bcl-2 protein expression was significantly up-regulated in the clones while no significant changes in Bax, Bcl-(xL), P glycoprotein and Hsp70 protein expression and no consistent changes in p53 expression were detected. We propose that recurrent growth factor starvation, which may occur in vivo when stromal function is damaged after intensive chemotherapy or bone marrow occupation by malignant cells, causes selection of drug resistant leukaemia cells that will expand when the growth factor supply recovers. PMID- 11870524 TI - Radiotherapy for early stage favourable breast cancers. PMID- 11870523 TI - Hypoxia-associated spontaneous pulmonary metastasis in human melanoma xenografts: involvement of microvascular hot spots induced in hypoxic foci by interleukin 8. AB - The aim of this study was to investigate whether tumour hypoxia and/or vascular hot spots promote the development of metastatic disease. The D-12 human melanoma xenograft line was used as a tumour model. Hypoxia and vascular hot spots were detected by immunohistochemistry using pimonidazole as a hypoxia marker and anti CD31 antibody to visualize endothelial cells. Vascular hot spots were found to be induced in hypoxic foci, owing to hypoxia-induced up-regulation of angiogenesis stimulatory factors. This effect was mediated by interleukin 8 and possibly also by vascular endothelial growth factor. Interleukin 8 positive foci showed a high degree of co-localization with hypoxic foci, as revealed by immunohistochemistry. The incidence of spontaneous pulmonary metastases was associated with the density of hypoxic foci, the density of interleukin 8 positive foci and the density of vascular hot spots in the primary tumour. Treatment with neutralizing antibody against interleukin 8 and/or vascular endothelial growth factor resulted in hypoxia-induced necrosis rather than hypoxia-induced vascular hot spots and inhibited metastasis. Our study suggests a cause-effect relationship between hypoxia and metastasis in cancer and hence an elevated probability of metastatic disease in patients having primary tumours characterized by high densities of hypoxic foci and vascular hot spots. PMID- 11870526 TI - NICE verdict on Temozolomide: where next? PMID- 11870527 TI - A rapid and systematic review of the effectiveness of temozolomide for the treatment of recurrent malignant glioma. AB - A rapid and systematic review of the effectiveness and cost-effectiveness of temozolomide in the treatment of recurrent malignant glioma was commissioned by the NHS HTA Programme on behalf of NICE. The full report has been published elsewhere. This paper summarizes the results for the effectiveness of temozolomide in people with recurrent glioblastoma multiforme and anaplastic astrocytoma. The review was conducted using standard systematic review methodology involving a systematic literature search, quality assessment of included studies with systematic data extraction and data synthesis. One randomized controlled trial and four uncontrolled studies were identified for inclusion. The key results were that temozolomide may increase progression-free survival but has no significant impact on overall length of survival. The main effect from temozolomide may have been in those patients who had not received any prior chemotherapy regimens, however further randomized controlled trials are required to confirm this suggestion. Temozolomide appears to produce few serious adverse effects and may also have a positive impact on health-related quality of life. Overall the evidence-base is weak and few strong conclusions can be drawn regarding the effectiveness of temozolomide. Large, well-designed randomized controlled trails conducted in a wider patient population are needed. PMID- 11870528 TI - Early prediction of treatment response to high-dose salvage chemotherapy in patients with relapsed germ cell cancer using [(18)F]FDG PET. AB - To assess the ability of [(18)F]fluorodeoxyglucose positron emission tomography for the early prediction of response in patients with relapsed metastatic germ cell tumours undergoing salvage high-dose chemotherapy. The role of positron emission tomography was compared with established means of tumour response assessment such as CT scans/MRI and serum tumour marker changes. In addition, positron emission tomography was compared with a current prognostic score which differentiates three prognostic groups with failure-free survival rates ranging from 5-50%. [(18)F]fluorodeoxyglucose uptake of metastases from germ cell tumours as well as CT scans and serum tumour marker were acquired after 2-3 cycles of induction chemotherapy but before the start of high-dose chemotherapy and CT scans/serum tumour marker were compared with the baseline examinations in 23 patients with relapsed germ cell tumours. To evaluate the validity of early response prediction by positron emission tomography, radiological monitoring and serum tumour marker decline, histopathologic response after resection of residual masses and/or the clinical course over 6 months after the end of treatment (relapse vs freedom of progression) were used. Overall, 10 patients (43%) achieved a marker-negative partial remission, three (13%) a marker-positive partial remission, five (22%) a disease stabilization and five (22%) progressed during treatment. Nine patients (39%) remained progression-free over 6 months following treatment, whereas 14 (61%) progressed. The outcome of high-dose chemotherapy was correctly predicted by positron emission tomography/CT scan/serum tumour marker in 91/59/48%. Eight patients with a favourably predicted outcome by CT scans plus serum tumour marker but a positive positron emission tomography prior to high-dose chemotherapy, failed treatment. This results in the following sensitivities/specificities for the prediction of failure of high-dose chemotherapy: positron emission tomography 100/78%; radiological monitoring 43/78%; serum tumour marker 15/100%. The positive and negative predictive values of positron emission tomography were 88 and 100%, respectively. As compared with the prognostic score, positron emission tomography was correctly positive in all patients of the three risk groups who failed treatment. In addition, a negative positron emission tomography correctly predicted a favourable outcome in the good and intermediate group. [(18)F]fluorodeoxyglucose positron emission tomography imaging can be used to assess response to chemotherapy in patients with relapsed germ cell tumours early in the course of treatment and may help to identify patients most likely to achieve a favourable response to subsequent high-dose chemotherapy. In patients with response to induction chemotherapy according to CT scans or serum tumour marker evaluation, positron emission tomography seems to add information to detect patients with an overall unfavourable outcome. It may also be a valuable addition to the prognostic model particularly in the good and intermediate group for further selection of patients who will profit from high dose chemotherapy. PMID- 11870529 TI - FDG-PET. A possible prognostic factor in head and neck cancer. AB - Previous studies have shown that high uptake of (18)F-fluoro-2-deoxy-glucose in head and neck cancer, as determined by the standardized uptake value on positron emission tomography scan, was associated with poor survival. The aim of this study was to confirm the association and to establish whether a high standardized uptake value had prognostic significance. Seventy-three consecutive patients with newly diagnosed squamous cell carcinoma of the head and neck underwent a positron emission tomography study before treatment. Age, gender, performance status tumour grade, stage, maximal tumour diameter and standardized uptake value were analyzed for their possible association with survival. The median standardized uptake value for all primary tumours was 7.16 (90% range 2.30 to 18.60). In univariate survival analysis the cumulative survival was decreased as the stage, tumour diameter and standardized uptake value increased. An standardized uptake value of 10 was taken as a cut-off for high and low uptake tumours. When these two groups were compared, an standardized uptake value >10 predicted for significantly worse outcome (P=0.003). Multivariate analysis demonstrated that an standardized uptake value >10 provided prognostic information independent of the tumour stage and diameter (P=0.002). We conclude that high FDG uptake (standardized uptake value>10) on positron emission tomography is an important marker for poor outcome in primary squamous cell carcinoma of the head and neck. Standardized uptake value may be useful in distinguishing those tumours with a more aggressive biological nature and hence identifying patients that require intensive treatment protocols including hyperfractionated radiotherapy and/or chemotherapy. PMID- 11870531 TI - CT-guided intratumoural administration of cisplatin/epinephrine gel for treatment of malignant liver tumours. AB - To analyze prospectively the interventional and clinical aspects of computed tomography-guided direct intratumoural injection of a novel chemotherapeutic administration and the parenchymal changes of tumour and necrosis in malignant liver tumours. Eight patients with 17 colorectal liver metastases were treated with a mean of 5.1 injections and nine patients with 13 hepatocellular carcinoma nodules with a mean of 3.1 treatments with computed tomography guided local applications of a novel cisplatin/epinephrine gel. This application provides a higher local and lower systemic drug concentration. Volumes of tumour and necrosis prior and after treatment were measured by computer generated volumetric analysis. Contrast enhanced studies verified pretherapeutic viable tumour volumes with a value of 77.4 ml in the metastases and 29.2 ml in the hepatocellular carcinoma nodules. Intratumoural drug application resulted in a significant increase of necrosis and a decrease in viable tumour volume to be 68.3 ml in metastases and 14.5 ml in hepatocellular carcinoma. Local therapy control rate for the follow up to 6 months was 38 and 71% for the group of metastases and hepatocellular carcinoma, respectively. Direct intratumoural injection of cisplatin/epinephrine injectable gel is a feasible and good tolerated method and results in the development of a statistically significant increase in necrosis in malignant liver tumours. For hepatocellular carcinoma a higher local therapy control rate compared to colorectal metastases can be reported. PMID- 11870530 TI - Accelerated hyperfractionation (AHF) compared to conventional fractionation (CF) in the postoperative radiotherapy of locally advanced head and neck cancer: influence of proliferation. AB - Based on the assumption that an accelerated proliferation process prevails in tumour cell residues after surgery, the possibility that treatment acceleration would offer a therapeutic advantage in postoperative radiotherapy of locally advanced head and neck cancer was investigated. The value of T(pot) in predicting the treatment outcome and in selecting patients for accelerated fractionation was tested. Seventy patients with (T2/N1-N2) or (T3-4/any N) squamous cell carcinoma of the oral cavity, larynx and hypopharynx who underwent radical surgery, were randomized to either (a) accelerated hyperfractionation: 46.2 Gy per 12 days, 1.4 Gy per fraction, three fractions per day with 6 h interfraction interval, treating 6 days per week or (b) Conventional fractionation: 60 Gy per 6 weeks, 2 Gy per fraction, treating 5 days per week. The 3-year locoregional control rate was significantly better in the accelerated hyperfractionation (88 +/- 4%) than in the CF (57+/- 9%) group, P=0.01 (and this was confirmed by multivariate analysis), but the difference in survival (60 +/- 10% vs 46 +/- 9%) was not significant (P=0.29). The favourable influence of a short treatment time was further substantiated by demonstrating the importance of the gap between surgery and radiotherapy and the overall treatment time between surgery and end of radiotherapy. Early mucositis progressed more rapidly and was more severe in the accelerated hyperfractionation group; reflecting a faster rate of dose accumulation. Xerostomia was experienced by all patients with a tendency to be more severe after accelerated hyperfractionation. Fibrosis and oedema also tended to be more frequent after accelerated hyperfractionation and probably represent consequential reactions. T(pot) showed a correlation with disease-free survival in a univariate analysis but did not prove to be an independent factor. Moreover, the use of the minimum and corrected P-values did not identify a significant cut off. Compared to conventional fractionation, accelerated hyperfractionation did not seem to offer a survival advantage in fast tumours though a better local control rate was noted. This limits the use of T(pot) as a guide for selecting patients for accelerated hyperfractionation. For slowly growing tumours, tumour control and survival probabilities were not significantly different in the conventional fractionation and accelerated hyperfractionation groups. A rapid tumour growth was associated with a higher risk of distant metastases (P=0.01). In conclusion, tumour cell repopulation seems to be an important determinant of postoperative radiotherapy of locally advanced head and neck cancer despite lack of a definite association between T(pot) and treatment outcome. In fast growing tumours accelerated hyperfractionation provided an improved local control but without a survival advantage. To gain a full benefit from treatment acceleration, the surgery-radiotherapy gap and the overall treatment time should not exceed 6 and 10 weeks respectively. PMID- 11870532 TI - Preoperative induction chemotherapy with cisplatin and irinotecan for pathological N(2) non-small cell lung cancer. AB - We conducted a phase I/II study to investigate whether the surgical resection after induction chemotherapy with cisplatin and irinotecan was feasible and could improve the treatment outcome for patients with pathological N(2) non-small cell lung cancer. Fifteen patients with stage IIIA non-small cell lung cancer having mediastinal lymph node metastases proved by mediastinoscopy were eligible. Both cisplatin (60 mg m(-2)) and irinotecan (50 mg m(-2)) were given on days 1 and 8. Patients received two cycles of chemotherapy after 3-4 weeks interval. Induction was followed by surgical resection in 4-6 weeks. Patients who had documented tumour regression after preoperative chemotherapy received two additional cycles of chemotherapy and other patients received radiotherapy postoperatively. After the induction chemotherapy, the objective response rate was 73%. All the 15 patients received surgical resection and complete resection was achieved in 11 (73%) patients. There was no operation-related death and one death due to radiation pneumonitis during postoperative radiotherapy. The median time from entry to final analysis was 46.5 months, ranging from 22 to 68 months. The 5-year survival rate was 40% for all the 15 patients and it was 55% for the 11 patients who underwent complete resection. We conclude that the surgical resection after induction chemotherapy with cisplatin and irinotecan is feasible, and associated with low morbidity and high respectability. PMID- 11870533 TI - Phase I study of the combination of losoxantrone and cyclophosphamide in patients with refractory solid tumours. AB - Losoxantrone is a DNA intercalator that was developed with the potential to replace anthracyclines. The recommended single agent dose of losoxantrone is 50 mg m(-2) every 3 weeks. We conducted a phase I study of losoxantrone and a fixed dose of cyclophosphamide on a q3 weekly schedule. Forty-nine patients were enrolled, of which 46 were evaluable for toxicity. The dose-limiting toxicity was neutropenia at the maximum tolerable losoxantrone dose of 45 mg m(-2). With granulocyte colony-stimulating factor support, significant further dose escalation of losoxantrone was achieved. Cardiotoxicity was seen with cumulative dosing. Pharmacokinetics of losoxantrone revealed linear kinetics and triphasic clearance, with significant interpatient variability. No objective responses were seen in this study. Neutropenia was dose-limiting in this combination with or without granulocyte colony-stimulating factor support. The recommended dose for further testing is cyclophosphamide 500 mg m(-2) followed by losoxantrone 95 mg m(-2) with granulocyte colony-stimulating factor support. PMID- 11870534 TI - Activation of AKT/PKB in breast cancer predicts a worse outcome among endocrine treated patients. AB - Akt/PKB is a serine/threonine protein kinase that regulates cell cycle progression, apoptosis and growth factor mediated cell survival in association with tyrosine kinase receptors. The protein is a downstream effector of erbB-2 with implications in breast cancer progression and drug resistance in vitro. We aimed to examine the role of Akt-1 in breast cancer patients, by determining whether the expression (Akt-1) and/or activation (pAkt) were related to prognostic markers and survival. The expression of erbB-2, heregulin beta 1 and Bcl-2 was also assessed by flow cytometry or immunohistochemistry. This study comprised 93 patients, aged <50 who were treated with tamoxifen and/or goserelin. We found that pAkt was associated with lower S-phase fraction (P=0.001) and the presence of heregulin beta 1-expressing stromal cells (P=0.017). Neither Akt-1 nor pAkt was related with other factors. Tumour cells-derived heregulin beta 1 was found mainly in oestrogen receptor negative (P=0.026) and node negative (P=0.005) cases. Survival analysis revealed that pAkt positive patients were more prone to relapse with distant metastasis, independently of S-phase fraction and nodal status (multivariate analysis; P=0.004). The results suggest that activation of Akt may have prognostic relevance in breast cancer. PMID- 11870535 TI - Dendritic cell density and activation status in human breast cancer -- CD1a, CMRF 44, CMRF-56 and CD-83 expression. AB - Low CD1a-positive putative dendritic cell numbers in human breast cancer has recently been described and may explain the apparent 'poor immunogenicity' previously reported in breast cancer. Little attention has been given to dendritic cell activation within the tumour microenvironment, which is another reason why the in-situ immune response may be severely deficient. We have therefore examined CD1a expression as a marker for dendritic cells, together with CMRF-44 and -56 as markers of dendritic cell activation status, in 40 human breast cancers. The results demonstrate few or no CD1a-positive putative dendritic cells and minimal or no expression of the dendritic cell activation markers. Both dendritic cell number and dendritic cell activation appear substantially deficient in human breast cancers, regardless of tumour histological grade. PMID- 11870537 TI - A high vascular count and overexpression of vascular endothelial growth factor are associated with unfavourable prognosis in operated small cell lung carcinoma. AB - It has been widely demonstrated that neo-angiogenesis and its mediators (i.e. vascular endothelial growth factor), represent useful indicators of poor prognosis in non small cell lung carcinoma. In order to verify whether neovascularization and vascular endothelial growth factor may be considered useful markers of clinical outcome also in the small cell lung cancer subgroup, we retrospectively investigated a series of 75 patients with small cell lung carcinoma treated by surgery between 1980 and 1990. Immunohistochemically detected microvessels and vascular endothelial growth factor expressing cells were significantly associated with poor prognosis, as well as with nodal status and pathological stage. In fact, patients whose tumours had vascular count and vascular endothelial growth factor expression higher than median value of the entire series (59 vessels per 0.74 mm(2) and 50% of positive cells, respectively), showed a shorter overall and disease-free survival (P=0.001, P=0.001; P=0.008, P=0.03). Moreover, the presence of hilar and/or mediastinal nodal metastasis and advanced stage significantly affected overall and disease free interval (P=0.00009, P=0.00001; P=0.0001, P=0.00001). At multivariate analysis, only vascular endothelial growth factor expression retained its influence on overall survival (P=0.001), suggesting that angiogenic phenomenon may have an important role in the clinical behaviour of this lung cancer subgroup. PMID- 11870536 TI - Expression of angiogenic factors predicts response to chemoradiotherapy and prognosis of oesophageal squamous cell carcinoma. AB - The ability to predict patients' responses to chemoradiotherapy by analyzing pre treatment biopsy specimens would be valuable for managing oesophageal squamous cell cancer. To this end, the expression of p53, thymidine phosphorylase and vascular endothelial cell growth factor was analyzed by immunohistochemistry in 52 patients with oesophageal squamous-cell cancer prior to chemoradiotherapy. Treatment consisted of radiotherapy (40 Gy) and 5 day-infusion of 5-Fluorouracil (500 mg m(-2) per day) combined with cisplatin (10 mg m(-2) per day). Following treatment, imaging and endoscopic reassessment was performed to establish treatment response. Thirty-one patients underwent radical surgery and 21 patients were treated with an additional 20 Gy of radiotherapy. Of the tumours studied, 58% were p53-positive, 40% thymidine phosphorylase-positive and 44% vascular endothelial cell growth factor-positive. A clinical response was observed in 36 patients (69%) and was negatively associated with thymidine phosphorylase expression (P=0.02) and vascular endothelial cell growth factor expression (P<0.001). However, the 5-year survival rate was significantly lower only in patients with vascular endothelial cell growth factor-positive tumours (P=0.037). Multivariate analysis identified vascular endothelial cell growth factor as a significant independent prognostic factor (P=0.0147). These results suggest that expression of angiogenic factors has predictive value for the treatment response and outcome of patients with oesophageal cancer. PMID- 11870538 TI - Hypermethylation of the hMLH1 gene promoter in solitary and multiple gastric cancers with microsatellite instability. AB - Human cancers with a high frequency microsatellite instability phenotype develop due to defects in DNA mismatch repair genes. Silencing of a DNA mismatch repair gene, hMLH1 gene, by promoter hypermethylation is a frequent cause of the microsatellite instability-H phenotype. Using methylation specific PCR we investigated the methylation status of the hMLH1 gene promoter in 17 solitary gastric cancers (12 microsatellite instability-H and five microsatellite stable tumours from 17 patients), and 13 multiple gastric cancers (eight microsatellite instability-H, one low frequency microsatellite instability-L and four microsatellite stable tumours from five patients) and also examined non-cancerous gastric mucosa both adjacent to and distant from each tumour. Expression of hMLH1 protein was evaluated by immunohistochemistry. All microsatellite instability-H tumours (20 out of 20) had evidence of methylation of hMLH1 promoter, whereas only one out of 10 microsatellite instability-L and microsatellite stable tumours did (P<0.0000005), and the methylation status correlated with hMLH1 protein expression (P<0.000003). Furthermore, methylation of the hMLH1 promoter was detected in 50% (6 out of 12) and 63% (5 out of 8) of non-cancerous gastric mucosa samples adjacent to, and in 33% (4 out of 12) and 40% (2 out of 5) of those obtained from distant portion of, solitary and multiple cancers with microsatellite instability-H. Thus both solitary and multiple gastric cancers with microsatellite instability-H have evidence of similar high levels of hMLH1 promoter hypermethylation in the surrounding non-cancerous tissue. Hypermethylation of the hMLH1 promoter occurs in non-cancerous gastric mucosa of microsatellite instability-H tumours and may increase the risk of subsequent neoplasia. PMID- 11870539 TI - c-erbB-2 is not a major factor in the development of colorectal cancer. AB - We have investigated c-erbB-2 protein expression in a large cohort of well characterized colorectal tumours, and in a subset of lymph node metastases. We have also evaluated a Val(655)Ile single nucleotide polymorphism, which is associated with an increased risk of breast cancer, in a subset of the colorectal cancer patients and in healthy control subjects. Immunohistochemical studies revealed that while 81.8% of tumours expressed c-erbB-2, in the majority of cases equivalent levels of c-erb-B2 were seen in adjacent normal mucosa. Colon tumours were significantly more likely to express c-erbB-2 than rectal tumours (P=0.015). Only 52.4% of the metastases displayed staining patterns concordant with their primary tumour, indicating that determination of c-erbB-2 protein in colorectal tumours cannot predict the status of lymph node metastases. PCR--RFLP analysis of the Val(655)Ile single nucleotide polymorphism demonstrated that allele frequencies were identical between colorectal cancer patients and a control group of Caucasian subjects (Ile=0.80 and Val=0.20 in each case), indicating that it is not related to the risk of developing colorectal cancer in this population. Furthermore, there was no relationship between c-erbB-2 protein expression and gene polymorphism (P=0.58). In terms of prognosis, no association was seen between either c-erbB-2 protein expression or the presence of the Val allele and patient survival (P>0.05 in each case), suggesting that c-erbB-2 is not a prognostic marker in colorectal cancer. PMID- 11870540 TI - Methylation of hMLH1 promoter correlates with the gene silencing with a region specific manner in colorectal cancer. AB - Microsatellite instability is present in over 80% of the hereditary non-polyposis colorectal carcinoma and about 15-20% of the sporadic cancer. Microsatellite instability is caused by the inactivation of the mismatch repair genes, such as primarily hMLH1, hMSH2. To study the mechanisms of the inactivation of mismatch repair genes in colorectal cancers, especially the region-specific methylation of hMLH1 promoter and its correlation with gene expression, we analysed microsatellite instability, expression and methylation of hMLH1 and loss of heterozygosity at hMLH1 locus in these samples. Microsatellite instability was present in 17 of 71 primary tumours of colorectal cancer, including 14 of 39 (36%) mucinous cancer and three of 32 (9%) non-mucinous cancer. Loss of hMLH1 and hMSH2 expression was detected in nine and three of 16 microsatellite instability tumours respectively. Methylation at CpG sites in a proximal region of hMLH1 promoter was detected in seven of nine tumours that showed no hMLH1 expression, while no methylation was present in normal mucosa and tumours which express hMLH1. However, methylation in the distal region was observed in all tissues including normal mucosa and hMLH1 expressing tumours. This observation indicates that methylation of hMLH1 promoter plays an important role in microsatellite instability with a region-specific manner in colorectal cancer. Loss of heterozygosity at hMLH1 locus was present in four of 17 cell lines and 16 of 54 tumours with normal hMLH1 status, while loss of heterozygosity was absent in all nine cell lines and nine tumours with abnormal hMLH1 status (mutation or loss of expression), showing loss of heterozygosity is not frequently involved in the inactivation of hMLH1 gene in sporadic colorectal cancer. PMID- 11870541 TI - Prognostic value of CCND1 gene status in sporadic breast tumours, as determined by real-time quantitative PCR assays. AB - The CCND1 gene, a key cell-cycle regulator, is often altered in breast cancer, but the mechanisms underlying CCND1 dysregulation and the clinical significance of CCND1 status are unclear. We used real-time quantitative PCR and RT-PCR assays based on fluorescent TaqMan methodology to quantify CCND1 gene amplification and expression in a large series of breast tumours. CCND1 overexpression was observed in 44 (32.8%) of 134 breast tumour RNAs, ranging from 3.3 to 43.7 times the level in normal breast tissues, and correlated significantly with positive oestrogen receptor status (P=0.0003). CCND1 overexpression requires oestrogen receptor integrity and is exacerbated by amplification at 11q13 (the site of the CCND1 gene), owing to an additional gene dosage effect. Our results challenge CCND1 gene as the main 11q13 amplicon selector. The relapse-free survival time of patients with CCND1-amplified tumours was shorter than that of patients without CCND1 alterations, while that of patients with CCND1-unamplified-overexpressed tumours was longer (P=0.011). Only the good prognostic significance of CCND1 unamplified-overexpression status persisted in Cox multivariate regression analysis. This study confirms that CCND1 is an ER-responsive or ER-coactivator gene in breast cancer, and points to the CCND1 gene as a putative molecular marker predictive of hormone responsiveness in breast cancer. Moreover, CCND1 amplification status dichotomizes the CCND1-overexpressing tumors into two groups with opposite outcomes. PMID- 11870542 TI - Co-transduction of Apaf-1 and caspase-9 highly enhances p53-mediated apoptosis in gliomas. AB - Mutation of the p53 gene plays a critical role in the development of cancer and response to cancer therapy. To analyze the mechanism of cancer development and to improve cancer therapy, it is important to assess which genes are downstream components of p53 in cancers, and whether the expression levels of these genes affect p53-mediated apoptosis. In this study, we transduced the wild type p53 gene along with the Apaf-1 and caspase-9 genes via adenovirus vectors into U251 and U-373MG glioma cells harbouring a mutated p53, and evaluated the degree of apoptosis. Co-induction of Apaf-1 and caspase-9 genes highly enhanced p53 mediated apoptosis in glioma cells. Induction of wild type p53 enhanced the expression levels of Bax, p21/WAF1, and Fas protein. To determine which gene is activated by wild type p53 induction and, in turn, activates Apaf-1 and caspase 9, we transduced the Bax, p21/WAF1 or Fas gene via adenovirus vector to U251 cells to achieve a similar expression level as that induced by the Adv for p53 in U251 cells. U251 cells transduced with Fas concomitant with the Apaf-1 and caspase-9 genes underwent drastic apoptosis. This suggests that induction of wild type p53 upregulates Fas, which in turn may play a role in the activation of Apaf 1 and caspase-9. These results are important for analyzing the mechanism of tumour development and for predicting the therapeutic effect of p53 replacement gene therapy in a particular patient. PMID- 11870544 TI - Inhibition of all-TRANS-retinoic acid metabolism by R116010 induces antitumour activity. AB - All-trans-retinoic acid is a potent inhibitor of cell proliferation and inducer of differentiation. However, the clinical use of all-trans-retinoic acid in the treatment of cancer is significantly hampered by its toxicity and the prompt emergence of resistance, believed to be caused by increased all-trans-retinoic acid metabolism. Inhibitors of all-trans-retinoic acid metabolism may therefore prove valuable in the treatment of cancer. In this study, we characterize R116010 as a new anticancer drug that is a potent inhibitor of all-trans-retinoic acid metabolism. In vitro, R116010 potently inhibits all-trans-retinoic acid metabolism in intact T47D cells with an IC(50)-value of 8.7 nM. In addition, R116010 is a selective inhibitor as indicated by its inhibition profile for several other cytochrome P450-mediated reactions. In T47D cell proliferation assays, R116010 by itself has no effect on cell proliferation. However, in combination with all-trans-retinoic acid, R116010 enhances the all-trans-retinoic acid-mediated antiproliferative activity in a concentration-dependent manner. In vivo, the growth of murine oestrogen-independent TA3-Ha mammary tumours is significantly inhibited by R116010 at doses as low as 0.16 mg kg(-1). In conclusion, R116010 is a highly potent and selective inhibitor of all-trans retinoic acid metabolism, which is able to enhance the biological activity of all trans-retinoic acid, thereby exhibiting antitumour activity. R116010 represents a novel and promising anticancer drug with an unique mechanism of action. PMID- 11870543 TI - Analyses of apoptotic regulators CASP9 and DFFA at 1P36.2, reveal rare allele variants in human neuroblastoma tumours. AB - The genes encoding Caspase-9 and DFF45 have both recently been mapped to chromosome region 1p36.2, that is a region alleged to involve one or several tumour suppressor genes in neuroblastoma tumours. This study presents an update contig of the 'Smallest Region of Overlap of deletions' in Scandinavian neuroblastoma tumours and suggests that DFF45 is localized in the region. The genomic organization of the human DFF45 gene, deduced by in-silico comparisons of DNA sequences, is described for the first time in this paper. In the present study 44 primary tumours were screened for mutation by analysis of the genomic sequences of the genes. In two out of the 44 tumours this detected in the DFFA gene one rare allele variant that caused a non-polar to a polar amino acid exchange in a preserved hydrophobic patch of DFF45. One case was hemizygous due to deletion of the more common allele of this polymorphism. Out of 194 normal control alleles only one was found to carry this variant allele, so in respect of it, no healthy control individual out of 97 was homozygous. Moreover, our RT-PCR expression studies showed that DFF45 is preferably expressed in low-stage neuroblastoma tumours and to a lesser degree in high-stage neuroblastomas. We conclude that although coding mutations of Caspase-9 and DFF45 are infrequent in neuroblastoma tumours, our discovery of a rare allele in two neuroblastoma cases should be taken to warrant further studies of the role of DFF45 in neuroblastoma genetics. PMID- 11870545 TI - Expression of uncoupling proteins-1, -2 and -3 mRNA is induced by an adenocarcinoma-derived lipid-mobilizing factor. AB - The abnormalities of lipid metabolism observed in cancer cachexia may be induced by a lipid-mobilizing factor produced by adenocarcinomas. The specific molecules and metabolic pathways that mediate the actions of lipid-mobilizing factor are not known. The mitochondrial uncoupling proteins-1, -2 and -3 are suggested to play essential roles in energy dissipation and disposal of excess lipid. Here, we studied the effects of lipid-mobilizing factor on the expression of uncoupling proteins-1, -2 and -3 in normal mice. Lipid-mobilizing factor isolated from the urine of cancer patients was injected intravenously into mice over a 52-h period, while vehicle was similarly given to controls. Lipid-mobilizing factor caused significant reductions in body weight (-10%, P=0.03) and fat mass (-20%, P<0.01) accompanied by a marked decrease in plasma leptin (-59%, P<0.01) and heavy lipid deposition in the liver. In brown adipose tissue, uncoupling protein-1 mRNA levels were elevated in lipid-mobilizing factor-treated mice (+96%, P<0.01), as were uncoupling proteins-2 and -3 (+57% and +37%, both P<0.05). Lipid-mobilizing factor increased uncoupling protein-2 mRNA in both skeletal muscle (+146%, P<0.05) and liver (+142%, P=0.03). The protein levels of uncoupling protein-1 in brown adipose tissue and uncoupling protein-2 in liver were also increased with lipid-mobilizing factor administration (+49% and +67%, both P=0.02). Upregulation by lipid-mobilizing factor of uncoupling proteins-1, -2 and -3 in brown adipose tissue, and of uncoupling protein-2 in skeletal muscle and liver, suggests that these uncoupling proteins may serve to utilize excess lipid mobilized during fat catabolism in cancer cachexia. PMID- 11870547 TI - A cyclin-dependent kinase inhibitor (p21(WAF1/CIP1)) affects thymidine incorporation in human liver cancer cells. AB - p21(WAF1/CIP1) is a universal cyclin-dependent kinase inhibitor. To investigate the role of p21(WAF1/CIP1) in proliferation of human liver cancer cells, we examined the expression of p53, p21(WAF1/CIP1), cdk2 and cdk4 expression in two human liver cancer cell lines (HepG2 and PLC/PRF/5 cells). The effects of p21(WAF1/CIP1) on [(3)H]thymidine incorporation and cdks were also examined in these cells. HepG2 cells expressed all these proteins with lower level of cdk4. PLC/PRF/5 cells expressed the other proteins except p21(WAF1/CIP1). Transfection of p21(WAF1/CIP1) gene inhibited [(3)H]thymidine incorporation of both cells with different extent. Although the transfection of p21(WAF1/CIP1) did not affect cdk2 and cdk4 expression, it did reduce cdk2 kinase activity by 20%. These results suggest that: (a) p21(WAF1/CIP1) involved in DNA synthesis of human liver cancer cells; (b) p21(WAF1/CIP1) could be a target gene for the treatment of human hepatocellular carcinoma. PMID- 11870546 TI - Variation in mitochondrial function in hypoxia-sensitive and hypoxia-tolerant human glioma cells. AB - We have shown previously that human glioblastoma multiforme cells vary in their ability to survive under hypoxic conditions. Under oxygen limiting conditions, hypoxia-tolerant cells decrease their oxygen consumption rate whereas hypoxia sensitive cells continue to consume oxygen at a relatively steady rate until the oxygen supply becomes exhausted. We now show that hypoxia-tolerant and hypoxia sensitive cells exhibit distinct patterns of mitochondrial function in response to hypoxic challenge. Hypoxia-tolerant cell lines retain stable mitochondrial membrane potential and ATP concentration when incubated under oxygen limiting conditions. In addition, hypoxia-tolerant cell lines are consistently more sensitive to a wide spectrum of inhibitors of mitochondrial function than are hypoxia-sensitive cells. In contrast, the hypoxia-sensitive cells are unable to maintain stable mitochondrial membrane potential and ATP levels when incubated at reduced oxygen tension. These results demonstrate significant differences in the mitochondrial function between these two phenotypes and reinforce previous data that suggest a regulatory role for mitochondria in the development of hypoxia tolerance. PMID- 11870548 TI - Interaction of cimetidine with P450 in a mouse model of hepatocarcinogenesis initiation. AB - Many drugs and xenobiotics are lipophilic and they should be transformed into more polar water soluble compounds to be excreted. Cimetidine inhibits cytochrome P450. The aim of this study was to investigate the preventive and/or reversal action of cimetidine on cytochrome P450 induction and other metabolic alterations provoked by the carcinogen p-dimethylaminoazobenzene. A group of male CF1 mice received a standard laboratory diet and another group was placed on dietary p dimethylaminoazobenzene (0.5% w w(-1). After 40 days of treatment, animals of both groups received p-dimethylaminoazobenzene and two weekly doses of cimetidine (120 mg kg(-1), i.p.) during a following period of 35 days. Cimetidine prevented and reversed delta-aminolevulinate synthetase induction and cytochrome P450 enhancement provoked by p-dimethylaminoazobenzene. However, cimetidine did not restore haem oxygenase activity decreased by p-dimethylaminoazobenzene. Enhancement in glutathione S-transferase activity provoked by p dimethylaminoazobenzene, persisted in those animals then treated with cimetidine. This drug did not modify either increased lipid peroxidation or diminution of the natural antioxidant defence system (inferred by catalase activity) induced by p dimethylaminoazobenzene. In conclusion, although cimetidine treatment partially prevented and reversed cytochrome P450 induction, and alteration on haem metabolism provoked by p-dimethylaminoazobenzene AB, it did not reverse liver damage or lipid peroxidation. These results further support our hypothesis on the necessary existence of a multiple biochemical pathway disturbance for the onset of hepatocarcinogenesis initiation. PMID- 11870549 TI - Extracellular nucleotides inhibit growth of human oesophageal cancer cells via P2Y(2)-receptors. AB - Extracellular ATP is known to inhibit growth of various tumours by activating specific purinergic receptors (P2-receptors). Since the therapy of advanced oesophageal cancer is unsatisfying, new therapeutic approaches are mandatory. Here, we investigated the functional expression and potential antiproliferative effects of P2-purinergic receptors in human oesophageal cancer cells. Prolonged incubation of primary cell cultures of human oesophageal cancers as well as of the squamous oesophageal cancer cell line Kyse-140 with ATP or its stable analogue ATP gamma S dose-dependently inhibited cell proliferation. This was due to both an induction of apoptosis and cell cycle arrest. The expression of P2 receptors was examined by RT-PCR, immunocytochemistry, and [Ca(2+)](i)-imaging. Application of various extracellular nucleotides dose-dependently increased [Ca(2+)](i). The rank order of potency was ATP=UTP>ATP gamma S>ADP=UDP. 2 methylthio-ATP and alpha,beta-methylene-ATP had no effects on [Ca(2+)](i). Complete cross-desensitization between ATP and UTP was observed. Moreover, the phospholipase C inhibitor U73122 dose-dependently reduced the ATP triggered [Ca(2+)](i) signal. The pharmacological features strongly suggest the functional expression of G-protein coupled P2Y(2)-receptors in oesophageal squamous cancer cells. P2Y(2)-receptors are involved in the antiproliferative actions of extracellular nucleotides. Thus, P2Y(2)-receptors are promising target proteins for innovative approaches in oesophageal cancer therapy. PMID- 11870551 TI - Photochemical disruption of endocytic vesicles before delivery of drugs: a new strategy for cancer therapy. AB - The development of methods for specific delivery of drugs is an important issue for many cancer therapy approaches. Most of macromolecular drugs are taken into the cell through endocytosis and, being unable to escape from endocytic vesicles, eventually are degraded there, which hinders their therapeutic usefulness. We have developed a method, called photochemical internalization, based on light induced photochemical reactions, disrupting endocytic vesicles specifically within illuminated sites e.g. tumours. Here we present a new drug delivery concept based on photochemical internalization-principle -- photochemical disruption of endocytic vesicles before delivery of macromolecules, leading to an instant endosomal release instead of detrimental stay of the molecules in endocytic vesicles. Previously we have shown that illumination applied after the treatment with macromolecules substantially improved their biological effect both in vitro and in vivo. Here we demonstrate that exposure to light before delivery of protein toxin gelonin improves gelonin effect in vitro much more than light after. However, in vitro transfection with reporter genes delivered by non-viral and adenoviral vectors is increased more than 10- and six-fold, respectively, by both photochemical internalization strategies. The possible cellular mechanisms involved, and the potential of this new method for practical application of photochemical internalization concept in cancer therapy are discussed. PMID- 11870552 TI - Endothelial hyperplasia: an important indicator of actual angiogenesis. PMID- 11870553 TI - (99m)Tc-labelled Stealth liposomal doxorubicin (Caelyx) in glioblastomas and metastatic brain tumours. PMID- 11870550 TI - Stabilization of breast cancer xenograft tumour neovasculature by angiopoietin-1. AB - Angiopoietin-1 is a promoter of physiological vasculogenesis and angiogenesis because it induces vascular branching and smooth muscle recruitment to newly formed blood vessels. However, angiopoietin-1 expression in tumours appears to be uncommon, and angiopoietin-1 overexpression in cancer cells has been reported to lead to inhibition of xenograft tumour growth. We report here that angiopoietin-1 overexpression resulted in stabilization of tumour edge-associated blood vessels, as it prevented vessel dilation and dissociation of smooth muscle cells from existing vessels. In addition, angiopoietin-1 stimulated an infiltration of mesenchymal cells into the tumours, such that the coverage of microvessels by pericytes increased markedly, and the cancer cells were separated into small masses by the host stroma. The rates of both cancer cell proliferation and apoptosis decreased significantly in the presence of angiopoietin-1. Tie2, the receptor for angiopoietin-1, was found to be present in vascular smooth muscle cells in culture in addition to endothelial cells. These findings suggest that a vascular stabilization effect of angiopoietin-1 accounts for the inhibition of tumour growth. PMID- 11870556 TI - [Special topic issue: hematology/oncology]. PMID- 11870557 TI - ["Be smart--dont' start" campaign to prevent children from starting to smoke: an analysis according to type of school they attend]. AB - BACKGROUND: The present study describes the evaluation of a primary smoking prevention programme called "be smart--don't start", left. The programme is carried out as a competition and classes that participate decide not to smoke for a period of 6 months. Classes that stay smoke-free for that period of time can win a number of attractive prizes. Aim of this study was to examine, whether the programme is effective in delaying the onset of smoking in adolescents from different types of school in Germany. METHODS: In the years 1998/1999 a control group study with repeated assessment was carried out. In the study, smoking status was assessed in 1677 pupils with a mean age of 12.8 years (SD = 0.97) on three occasions: prior to the beginning of the intervention, after the intervention and 6 months after the end of the intervention. Pupils came from four different types of school in Germany. RESULTS: After the intervention, in the control group 13.1 % of the pupils reported to have smoked during the previous 4 weeks, compared to 7.6 % in the intervention group (OR = 1.84 (1.31 2.58), p < 0.001). In the follow-up assessment, 20,9 % in the control group and 16,4 % in the intervention group reported to have smoked (OR = 1.34 (1.03-1.75), p < 0.05). With regard to different school types, the effect on the "Gesamtschulen" (comprehensive school; high school) was the strongest. CONCLUSION: The results suggests an effect of the intervention on the delay of onset of smoking in pupils. PMID- 11870558 TI - [Paraneoplastic pityriasis rubra pilaris in metastatic adenocarcinoma without diagnosable primary]. AB - HISTORY AND ADMISSION FINDINGS: A 75-year-old woman, without a history of severe illness, developed an erythematosquamous skin disease on hands and forearms. After continued spreading of these cutaneous lesions, she was admitted to hospital, presenting with a generalised desquamating erythrodermia and marked pruritus. INVESTIGATIONS: Skin biopsy showed cornocutaneous signs with alternating ortho- and parakeratosis, typical for pityriasis rubra pilaris. Laboratory findings showed a chronic to acute inflammation with leukocytosis, granulocytosis in the differential blood count, raised C-reactive protein in a range from 54.7 to a maximum of 157.2 mg/l and a protein electrophoresis with elevated alpha1- and alpha2-fraction. TREATMENT AND COURSE: The erythrodermia only temporarily receded under systemic therapy with acitretin and prednisolone. The patient developed intermittent septic fever accompanied by reduction or loss of consciousness. The general condition of the patient worsened considerably. Out of a rapidly progressing pleural effusion malignant cells similar to adenocarcinoma were isolated. Because CA15-3 was elevated we conducted an extended search especially for a breast carcinoma, but found only pathologically enlarged axillary, mediastinal and abdominal lymph nodes in conventional X-ray, CT, ultrasound and endoscopic procedures. The patient died from paraneoplastic pulmonary embolism. At autopsy, the widespread metastatic dissemination from poorly differentiated adenocarcinoma was confirmed. A necrosis in the right breast containing tumour cell remnants could probably be regarded as the primary neoplasm. Immunohistochemically no definite proof of breast nor gastro-intestinal carcinoma could be found. CONCLUSIONS: This case presents a rare paraneoplastic cutaneous manifestation as pityriasis rubra pilaris triggered by a poorly differentiated adenocarcinoma. The primary neoplasm could not definitely be identified, neither pre nor post mortem. PMID- 11870559 TI - [WHO classification of acute myeloid leukaemia (AML) and the myelodysplastic syndrome (MDS)]. PMID- 11870560 TI - [Selective tyrosine kinase inhibitor imatinib (STI571) in haematological and oncological disease]. PMID- 11870561 TI - [Lectin-related use of mistletoe: an experimental form of treatment with preclinically demonstrated potential risks]. PMID- 11870562 TI - [Discussion with patients to obtain their informed consent for participating in randomized therapeutic trials]. PMID- 11870563 TI - [Detection of DPD-exon-14 skipping before 5-fluorouracil treatment?]. PMID- 11870564 TI - [What effects has the presence of prolymphocytes on prognosis and therapy of CLL? ]. PMID- 11870566 TI - [Current status of diagnosis and therapy of Whipple's disease]. PMID- 11870565 TI - [What can be the cause of elevated alpha-fetoprotein after hydrocele surgery?]. PMID- 11870567 TI - Clinical study comparing bleeding and nonbleeding rectal varices. AB - BACKGROUND AND STUDY AIMS: Although rectal varices constitute an important cause of lower digestive tract bleeding in patients with portal hypertension, the etiology and pathology of rectal varices remains controversial, and adequate treatment for rectal varices has yet to be established. In this study, we evaluated rectal varices to identify any common characteristics of varices which are susceptible to hemorrhage. PATIENTS AND METHODS: The patients included 40 individuals with rectal varices among 425 patients with portal hypertension who had been treated in our institution. We retrospectively examined patient data regarding underlying hepatic diseases, hepatic function and endoscopic findings with regard to varices. RESULTS: Bleeding from rectal varices occurred in 15 of the 40 patients. Although the prevalence of hemorrhage tended to increase with exacerbation of hepatic dysfunction, no significant differences were found. Similarly, although the incidence of hemorrhage tended to be somewhat higher in patients who had undergone any treatment for complicated esophageal varices than in patients who had not, no significant difference was found. The prevalence of hemorrhage from rectal varices significantly increased in rectal varices of more advanced form, and the prevalence was significantly higher in patients with positive "red color" sign. CONCLUSIONS: The prevalence of hemorrhage from rectal varices was significantly higher in patients with rectal varices of advanced form and/or with a positive "red color" sign. PMID- 11870568 TI - Effect of esophageal variceal band ligation on hemorrhoids, anorectal varices, and portal hypertensive colopathy. AB - BACKGROUND AND STUDY AIMS: Although it is known that obliteration of esophageal varices following endoscopic variceal band ligation results in an increase in the incidence of portal hypertensive gastropathy, the effect of variceal ligation on hemorrhoids, anorectal/colonic varices and portal hypertensive colopathy is not known. The aim of this study was to investigate the effect of endoscopic variceal band ligation on hemorrhoids, anorectal/colonic varices and portal hypertensive colopathy. PATIENTS AND METHODS: A total of 60 consecutive patients with cirrhosis of the liver and portal hypertension were prospectively studied. Upper gastrointestinal endoscopy and full-length colonoscopy were carried out before the patients underwent endoscopic variceal band ligation for esophageal varices and after obliteration of the varices following band ligation. RESULTS: Obliteration of esophageal varices by endoscopic variceal band ligation did not affect the incidence of hemorrhoids (37 % before and after), anorectal varices (40 % before and after), and portal hypertensive colopathy (57 % before and after). CONCLUSIONS: It is concluded that esophageal variceal band ligation does not affect the incidence of hemorrhoids, anorectal varices or portal hypertensive colopathy in patients with cirrhosis of the liver. PMID- 11870569 TI - Comparative study of portal hypertensive gastropathy in schistosomiasis and hepatic cirrhosis. AB - BACKGROUND AND STUDY AIMS: Portal hypertensive gastropathy (PHG) is a common finding in chronic liver disease, but it has not been evaluated in hepatosplenic schistosomiasis, a significant cause of noncirrhotic portal hypertension worldwide. Our study was aimed at comparing cirrhosis with hepatosplenic schistosomiasis with regard to the endoscopic and histological findings of PHG. PATIENTS AND METHODS: We included 43 patients with a history of upper digestive hemorrhage, 22 with cirrhosis and 21 with schistosomiasis, without previous surgical or endoscopic treatment. Upper digestive endoscopies with macrobiopsies of the gastric body were prospectively performed in all cases. RESULTS: The endoscopic signs of PHG were more prevalent in cirrhosis than schistosomiasis (81.8 % vs. 33.3 %; P < 0.05), and the mosaic pattern was the main finding. Histological abnormalities were evenly distributed. CONCLUSIONS: In agreement with other investigations, this study shows a lower prevalence of endoscopic findings of portal hypertensive gastropathy in noncirrhotic diseases. This difference cannot be explained by the underlying microscopic alterations, which were similar in both groups, suggesting that other factors must play a role in its pathogenesis. PMID- 11870570 TI - Endoscopic management of pancreatic pseudocyst: a long-term follow-up. AB - BACKGROUND AND STUDY AIMS: No studies with real long-term follow-up after endoscopic drainage of pancreatic pseudocysts are available. The present study was undertaken to investigate the long-term outcome of endoscopic management of pancreatic pseudocyst with a minimum follow-up of 2 years. PATIENTS AND METHODS: A total of 38 consecutive patients with pancreatic pseudocyst underwent endoscopic cystogastrostomy (n = 27), endoscopic cystoduodenostomy (n = 6) and transpapillary drainage (n = 5). Patients were monitored at 1 and 3 months after drainage, and finally between 24 and 80 months. Upper gastrointestinal endoscopy was done at 1 and 3 months after drainage while ultrasound was done at 3 months and at the end of follow-up. Endoscopic retrograde cholangiopancreatography (ERCP) was only done before cyst drainage if no cyst bulge was visible in the stomach or duodenum or if obstructive jaundice was present. RESULTS: Biliary pancreatitis was responsible for the pseudocyst in 19 cases while the remaining occurrences were caused by alcohol (n = 12) and trauma (n = 7). All forms of endoscopic drainage were effective in treating pancreatic pseudocyst and there was complete disappearance of the cyst within 3 months of drainage, irrespective of cause. Over a mean follow-up of 44.23 months (24 - 80 months). Three patients had symptomatic recurrences while three had asymptomatic recurrences; all had alcohol-induced pancreatitis. No recurrences were seen in the biliary pancreatitis and trauma group. All symptomatic recurrences were successfully managed with endoscopic cystogastrostomy and stenting. A massive bleed in one patient required surgery while stent block and cyst infection in three patients and perforation in one patient were managed conservatively. ERCP was done before cyst drainage in eight patients because there was no visible bulge into the stomach or duodenum (n = 5), or because obstructive jaundice was present (n = 3). In five patients ERCP revealed cyst duct communication. All these patients were managed by transpapillary drainage and there was only one asymptomatic recurrence in this group. CONCLUSION: Endoscopic management of pancreatic pseudocyst is quite an effective and safe mode of treatment in experienced hands. ERCP before the procedure is only required when the cyst does not bulge into gut lumen, for a decision about the feasibility of transpancreatic drainage. On long-term follow up, recurrences were seen only in the alcoholic pancreatitis group. In the biliary pancreatitis group, no recurrences were seen after cholecystectomy and removal of common bile duct (CBD) stones if present. No recurrences were seen in the trauma group. PMID- 11870571 TI - Endoscopic fluorescence observation of gastric mucosa after indomethacin treatment in rats. AB - BACKGROUND AND STUDY AIMS: The present authors have already reported that mucosal autofluorescence intensity was increased in gastric lesions induced by nonsteroidal anti-inflammatory drugs. The aim of this study was to clarify whether the observation of mucosal autofluorescence with a newly established endoscopic fluorescence analyzing system could help us to recognize indomethacin induced gastric lesion formation in vivo. MATERIALS AND METHODS: Gastric mucosal fluorescence intensity was measured time-sequentially after indomethacin treatment in rats, using a fluorescence endoscope system. The concentration of thiobarbituric acid-reactive substances in gastric mucosa was measured as an indicator of tissue lipid peroxidation. Fluorescent substances from rat stomachs were analyzed using high performance liquid chromatography. RESULTS: Treatment with indomethacin induced a time-dependent increase of fluorescence intensity. The concentration of thiobarbituric acid-reactive substances was also increased after the treatment. Pretreatment with radical scavenging reagent constrained the increase of both the fluorescence intensity and the concentration of thiobarbituric acid-reactive substances. The fluorescence products were coproporphyrin, protoporphyrin and mesoporphyrin. CONCLUSIONS: These findings indicate that the intensity of porphyrin fluorescence increased in gastric mucosal lesions induced by oxygen radicals. Endoscopic observation of mucosal fluorescence was shown to aid the sensitive and objective diagnosis of gastric injuries. PMID- 11870572 TI - Increased rectal wall thickness may predict relapse in ulcerative colitis: a pilot follow-up study by ultrasonographic colonoscopy. AB - BACKGROUND AND STUDY AIMS: Ulcerative colitis is a chronic inflammatory bowel disease with repeated remission and relapse, although the occurrence of relapse is difficult to predict. We performed a prospective study to determine whether there is a relationship in ulcerative colitis between the inflammatory changes identified by endoscopic ultrasonography (EUS) and relapse. PATIENTS AND METHODS: Participants were 23 ulcerative colitis patients who had not suffered a relapse for 1 month, with a Seo activity index less-than-or-equal 150 and Baron grade 1 at colonoscopy. The thickness of the first to the third layer of the rectal wall in these patients was measured by EUS at the start of the study. They were subsequently followed up for 1 year to record any relapses, defined as having a Seo activity index > 150 and Baron grade greater-than-or-equal 2 at colonoscopy. RESULTS: Relapse occurred in eight patients. The mean activity index of the relapsed patients was 187.3 (95 % confidence interval (CI), 166.4 - 208.2) at the end of the study. The thickness of the first to the third layer of the rectal wall, as evaluated by EUS at the beginning of the study, was significantly larger in the relapse group (mean 2.73 mm, 95 % CI 2.13 - 3.33 mm) than in the non relapse group (1.79 mm; 1.56 - 1.99 mm; P = 0.0001). CONCLUSIONS: Catheter probe assisted endoluminal ultrasonography may predict the occurrence of relapse of ulcerative colitis. PMID- 11870573 TI - Selective occlusion with fibrin glue under fistuloscopy: seven cases of postoperative management for intractable complex fistulas. AB - BACKGROUND AND STUDY AIM: Fistula occlusion is not achieved in some fistulas with complex branches. To obtain early fistula closure in such cases, we insert a double-lumen catheter into each fistula branch, with the aid of a guide wire positioned using a small-caliber endoscope, and attempt selective infusion of fibrin glue. PATIENTS AND METHODS: Following removal of foreign bodies and necrotic granulation, we applied the selective occlusion method under fistuloscopic control to seven intractable external fistulas with complex branches, in which fistula closure had not been obtained by a simple occlusion method (SOM). All the fistulas were complex with more than two branches. RESULTS: Fistula occlusion was obtained within 2 weeks in six of the seven patients, and there has been no sign of recurrence over a follow-up period of 4 - 59 months (average 29.8 months). CONCLUSION: Selective occlusion under fistuloscopy is highly effective for intractable external fistulas with complex branches. PMID- 11870574 TI - The spectrum of portal hypertension in the gastrointestinal tract. PMID- 11870575 TI - Classification of colorectal polyps: guidelines for the endoscopist. PMID- 11870576 TI - Massive rectal bleeding from a Dieulafoy lesion in the rectum: successful endoscopic clipping. PMID- 11870577 TI - Endosonographic misdiagnosis of tumor recurrence after surgery for malignant GIST. PMID- 11870579 TI - Percutaneous endoscopic jejunostomy for decompression in an infant with short bowel syndrome. PMID- 11870580 TI - Rectal leiomyoma with fibromuscular obliteration mimicking adematous lesion. PMID- 11870578 TI - Fast-growing gastric inflammatory pseudotumor: a rare manifestation of peptic ulcer disease. PMID- 11870581 TI - Classic in neuropediatrics. PMID- 11870582 TI - Cerebral dysplasias: reviewed from a neurophysiological perspective. AB - Improvements in neuroimaging techniques have highlighted the role of cerebral dysplasia in developmental disorders and in epilepsy. The current classification is founded on the presumed timing of the first insult as deduced from the imaging appearances. Although clinical neurophysiological findings are often non specific, several unusual patterns have been reported in association with certain types of malformation. These include the high amplitude rhythmic activities of lissencephaly, and focal rhythmic spikes. In this general review, we describe the electroence-phalographic and evoked potential findings in a wide range of cerebral dysplasias, including our personal experience of rare conditions such as Ramer-Lin syndrome. We focus on their role in assessment and on potential insights into pathophysiology. PMID- 11870583 TI - Neonatal diffusion-weighted MR imaging: relation with histopathology or follow-up MR examination. AB - PURPOSE: Diffusion-weighted imaging (DWI) has become a standard method for early evaluation of stroke in adults, but its value in neonates is less well established. In this study neonatal DWI was compared with histopathology in those patients who died, or with sequelae seen on a second MR in the surviving neonates. PATIENTS AND METHODS: DWI was performed in 2 groups. Group 1: seven neonates who died and had a post-mortem ex-amination (perinatal asphyxia [n=5], symptomatic hypoglycemia [n= 11, periventricular leukomalacia [n= 1]). Group 2: six surviving neonates with a second MR examination at three months of age (perinatal asphyxia [n= 21, neonatal stroke[n= 3], meningo-encephalitis [n= 1]). RESULTS: In group l neonatal DWI showed more extensive involvement than conventional MRI in 6 out of 7 patients. These changes were less extensive,however, than seen post-mortem by histopathology in 5 out of 7. In group 2 neonatal DWI showed more extensive involvement than conventional MRI in 2 out of 6; 4 out of 6, however, showed less extensive cystic evolution on follow up MRI at 3 months than expected from neonatal imaging. CONCLUSION: There was a good relation between hyperintense areas on DWI and areas of cytotoxic edema and neuronal damage on histopathology. In the survivors a second MRI showed cystic evolution in all, but the volume of the cysts was smaller than expected on the basis of the neonatal DWI findings. PMID- 11870584 TI - Gender-specific occurrence of West syndrome in patients with pyruvate dehydrogenase complex deficiency. AB - Pyruvate dehydrogenase complex (PDHC) deficiency, a major cause of congenital lactic acidemia in children, usually is complicated by seizures, and, in some patients, West syndrome has occurred. We diagnosed 60 patients with PDHC deficiency, including equal numbers of affected males and females. We studied the clinical features in 10 patients with West syndrome caused by PDHC deficiency, and examined the relation to the mutation of the E(1)alpha subunit, representing the great majority of PDHC deficiencies. Among 30 boys and 30 girls with PDHC deficiency,1 boy and 9 girls had West syndrome, even though overall West syndrome shows a slight male preponderance. Therefore, West syndrome associated with PDHC deficiency occurred in 9 of 30 female patients (33%), but in only 1 of 30 male patients (3%). The frequency of West syndrome in patients with PDHC deficiency was significantly higher in females than in males(p<0.05). Lactate concentrations in blood and CSF should be measured in female patients with West syndrome as a screening test for PDHC deficiency, because of gender-specific occurrence of West syndrome caused by PDHC deficiency. PMID- 11870585 TI - Characteristics of breathing abnormality in Leigh and its overlap syndromes. AB - In this report we describe the respiratory patterns of six patients with Leigh syndrome, including two individual cases with accompanying clinical phenotypes of Alpers disease and mitochondrial encephalopathy with ragged red fibers. In five cases where sleep apnea was monitored, each one showed isolated or post-sigh central apnea, hiccup, apneusis-like breathing and obstructive apnea in various combinations. The remaining patient with Alpers/Leigh overlap syndrome showed an apneusis-like pattern of dyspnea. The sleep structure was examined in three patients. Two patients with brainstem lesions showed a decrease in the deep sleep stages and an absence of REM sleep. Medullary lesions were found in four patients by magnetic resonance imaging or at autopsy and involved predominantly the dorsal respiratory group (DRG) of medullary neurons. The role of DRG lesions in the pathophysiology of respiratory symptoms in Leigh syndrome is discussed. PMID- 11870586 TI - Glial fibrillary acidic protein and neurofilament in children with cerebral white matter abnormalities. AB - Glial fibrillary acidic protein (GFAP) is the major structural protein of the intermediate filaments found in glial cells. Increased levels in the cerebrospinal fluid (CSF) have been found to indicate gliosis. Neurofilament (NFL) is a structural element of neurons, mainly found in large myelinated axons. Its presence in the CSF has been suggested to reflect destruction of axons. The aim of this study was to see if GFAP and NFL in the CSF of children with neurological disabilities and an abnormal signal on magnetic resonance imaging (MRI) of the cerebral white matter could be used to clarify the underlying neuropathology. The potential of GFAP and NFL to differentiate a progressive disease from a stationary disorder was investigated, as was the correlation with disability and clinical findings. CSF from 26 children, eleven with progressive and 15 with non-progressive disorders, was analysed. GFAP was increased in all, interpreted to reflect gliosis. NFL was elevated in seven and considered to indicate ongoing neuroaxonal damage as all but one patient were found to have a progressive disease. GFAP did not differentiate between progressive and non progressive disorders, although low levels were found in stationary and high levels in progressive disorders. The severity of the disability correlated with the NFL levels, but not with the concentration of GFAP. PMID- 11870588 TI - Early prediction of outcome with aEEG in preterm infants with large intraventricular hemorrhages. AB - BACKGROUND: The electrocortical background contains prognostic information in full-term asphyxiated newborn infants already during the first postnatal hours. In preterm infants with intra-ventricular hemorrhages (IVH) the background activity in EEG and amplitude-integrated EEG (aEEG) is depressed during the first days of life, and the extent of the depression correlates with the degree of IVH. However, it has not been previously evaluated whether very early aEEG can predict later outcome also in pre-term infants. OBJECTIVE: To investigate if early prediction of outcome is possible from aEEG in preterm infants with large IVH. METHODS: aEEG recordings from the first postnatal week were investigated in 64 preterm infants with IVH grade III - IV. For every 24-hour period the aEEG background pattern was classified, and the maximum and minimum numbers of bursts/h, respectively,were counted. Outcome was divided into three categories: died (n = 36), survived (n = 28) with "poor" outcome, i.e., severe cerebral palsy and not able to walk and/or mental retardation (n = 8), and survived with "fair" outcome, i.e., healthy or mild cerebral palsy (n = 19). One surviving child was lost in the follow-up. RESULTS: There were significant differences in maximum bursts/h (MaxB) at 0-24 hours (p = 0.033), 24-48 hours (p = 0.011), 48-72 hours (p=0.049) and 72-96 hours (p=0.032), respectively, between the infants who died and the surviving infants. At 24-48 hours the median (range) MaxB in the surviving infants with "fair" outcome was 156 (103-179) versus 102 (73-156) in the surviving infants with "poor" outcome (p = 0.002). With the assumption that MaxB < 130 was predictive of death or survival with "poor" outcome, 68 % and 78% of infants were correctly predicted at 0-24 hours and 24-48 hours, respectively. CONCLUSIONS: This study shows that outcome may be predicted with aEEG already during the first days of life in preterm infants with large IVH. The findings should be confirmed in prospective studies since they may have clinical implications if specific medical interventions become available. PMID- 11870587 TI - A new subtype of a congenital disorder of glycosylation (CDG) with mild clinical manifestations. AB - A boy with an unspecific symptomatology consisting of mental retardation, strabismus, hypotonia and mild ataxia was diagnosed with a congenital disorder of glycosylation (CDG). Neither cerebellar atrophy nor dysmorphic features were present. The serum transferrin band pattern obtained by isoelectric focusing(IEF) showed a strongly elevated disialotransferrin band together with only slightly elevated asialotransferrin, thus a type I pattern. This is a new CDG classified CDG-x since CDG-la, -b, -c, -d and -e were excluded. Quantitative differences to the type 1 pattern of a CDG-la patient with a moderate to severe course were confirmed by densitometric evaluation of the gels and by SDS gel electrophoresis. Liver biopsy showed lysosomal inclusions suggesting a pre-Golgi defect. This patient's case supports the approach to include isoelectric focusing of serum transferrin in the diagnostic work-up of patients with unexplained symptoms. PMID- 11870589 TI - Autosomal recessive polymicrogyria with infantile spasms and limb deformities. AB - We describe two siblings, a girl and a boy, aged 4 and 2 years and 10 months respectively, born from non-consanguineous parents,with diffuse polymicrogyria, lower limb deformities, infantile spasms and developmental delay. Spasms had a good outcome under antiepileptic drug treatment. Clinical and imaging features were of identical severity in both siblings. Muscle biopsy,creatine kinase, metabolic investigations and chromosomal analysis were normal. This combination of anatomo-clinical features and their occurrence in siblings of both sexes suggests an autosomal recessive malformation syndrome. PMID- 11870590 TI - Congenital cervical spinal atrophy: an intrauterine hypoxic insult. AB - We present two patients with congenital cervical spinal atrophy who were born at 37 and 33 weeks of gestation. Both patients were unrelated and had no family history of neuromuscular diseases. They presented at birth with arthrogryposis multiplex and symmetrical severe muscle weakness and wasting confined to the upper extremities. There was no sensory or bulbar symptom. Electromyography showed polyphasic and fast-firing units in the proximal muscles of the upper extremities. With the evidence of chronic denervation and re-innervation, we speculate that this static condition is most likely due to circulatory insufficiency causing anterior horn cell ischemia during the latter part of the first trimester. PMID- 11870591 TI - The pathogenesis of hydrocephalus in inborn errors of the single carbon transfer pathway. PMID- 11870592 TI - Protecting subjects' interests in genetics research. AB - Biomedical researchers often assume that sponsors, subjects, families, and disease-associated advocacy groups contribute to research solely because of altruism. This view fails to capture the diverse interests of many participants in the emerging research enterprise. In the past two decades, patient groups have become increasingly active in the promotion and facilitation of genetics research. Simultaneously, a significant shift of academic biomedical science toward commercialization has occurred, spurred by U.S. federal policy changes. The concurrent rise in both the roles that subjects play and the commercial interests they have presents numerous ethical challenges. We examine the interests of different research participants, finding that these interests are not addressed by current policies and practices. We conclude that all participants should be given a voice in decisions affecting ownership, access to, and use of commercialized products and services, and that researchers and institutions should negotiate issues relating to control of research results and the sharing of benefits before the research is performed. PMID- 11870595 TI - Squamotympanic fissure in the Ngandong and Sambungmacan hominids: a reply to Delson et al. AB - In the present study the course of the squamotympanic fissure of the mandibular fossa is examined in several fossil hominid specimens. While previous work by several authors had found that the course of this fissure was potentially autapomorphic for the Ngandong and Sambungmacan fossil samples from Indonesia, a more recent study performed by Delson et al. (Anat Rec 2001; 262:380--397) reported that a number of other fossils, including OH 9, Sangiran 4 and 17, and KNM WT 15000 were similar to the Ngandong and Sambungmacan specimens in the expression of this feature. To test these findings, those specimens purported to share similarities with Ngandong and Sambungmacan were examined. The results of this study indicate clearly differing morphologies between the Ngandong and Sambungmacan fossils, on the one hand, and the four African and Indonesian fossils named by Delson et al. (2001). Hence, the course of the squamotympanic fissure remains a potentially autapomorphic feature separating the Ngandong and Sambungmacan samples from the remaining specimens allocated to Homo erectus. PMID- 11870597 TI - Original histologic findings in arteries of the right ventricle papillary muscles in human hearts. AB - In this study we describe original histologic findings of the right ventricle papillary muscle (PM) arteries in people under 30 years old. We examined 666 samples taken from the tip, mid-portion, and base of the PM in 56 males and 55 females, as well as samples from the rest of the right ventricle. The amount of smooth muscle cell (SMC) fibers in the tunica media (TM) led to their division into three groups: Group 1: 351 samples (53%); normal amount, normal lumen. The amount of SMCs increased from the tip (20%) to the base (48%). Group 2: 226 samples (34%); mild to moderately increased amount of SMCs, with narrowness, eccentric displacement, and uneven lumen shape. They decreased from the tip (42%) to the base (23%). Group 3: 89 samples (13%), with abundant SMCs that duplicated the arterial size, contrary to the other two groups. Their shape was round and their extremely narrow, centrally located lumen had a round or oval shape. These changes were restricted only to PM arteries and decreased from the tip (65%) to the mid-portion (35%). This type of artery predominated compared to the other two groups, probably because of the narrow lumen. No inflammatory reaction or chronic ischemic changes were found in the PM and its arteries. The SMC changes in groups 2 and 3 were found in subjects older than 2 months. The above findings will provide anatomists, cardiologists, and physiologists with valuable knowledge. PMID- 11870593 TI - A new susceptibility locus for autosomal dominant pancreatic cancer maps to chromosome 4q32-34. AB - Pancreatic cancer is the fifth leading cause of cancer death in the United States. Nearly every person diagnosed with pancreatic cancer will die from it, usually in <6 mo. Familial clustering of pancreatic cancers is commonly recognized, with an autosomal dominant inheritance pattern in approximately 10% of all cases. However, the late age at disease onset and rapid demise of affected individuals markedly hamper collection of biological samples. We report a genetic linkage scan of family X with an autosomal dominant pancreatic cancer with early onset and high penetrance. For the study of this family, we have developed an endoscopic surveillance program that allows the early detection of cancer and its precursor, before family members have died of the disease. In a genomewide screening of 373 microsatellite markers, we found significant linkage (maximum LOD score 4.56 in two-point analysis and 5.36 in three-point analysis) on chromosome 4q32-34, providing evidence for a major locus for pancreatic cancer. PMID- 11870598 TI - Temporomandibular joint in miniature pigs: anatomy, cell replication, and relation to loading. AB - The mechanical environment is a regulator of growth and adaptation of the musculoskeletal system, including joints. Although pigs (Sus scrofa) are used frequently as models for temporomandibular joint (TMJ) dysfunction, no systematic description of microanatomy exists for this species. We injected the thymidine analog 5-bromo-2'-deoxyuridine (BrdU) into 10- to 11-month-old miniature pigs that were undergoing measurements of TMJ bone strain. Ten hr later, the animals were sacrificed and their heads were perfused. Histological sections were used to map the distribution of replicating cells. Additional observations were made on gross dissections of jaw joints obtained from an abattoir. The pig TMJ is better supported than that of humans laterally and medially, but more vulnerable posteriorly. The posterior attachment area of the intra-articular disc is fibro fatty rather than vascular, as in humans. Cartilage lines the articular eminence as well as the condylar surface. At the posterosuperior region of the condyle, the cartilage ends abruptly and is replaced by an invaginating, actively replicating periosteum. Almost all of the BrdU-labeled cells resided in the prechondroblastic zones. The condyle had more replicating cells than did the eminence (P < 0.02), but lateral and medial locations did not differ in either element. In sagittal sections, the condyle had more replicating cells posteriorly (P < 0.001), but no A-P differences were seen in the eminence. Comparisons of these data with data on bone strain indicate that increased loading is negatively associated with cell replication. PMID- 11870599 TI - Morphologic changes in the TMJ following splint wear. AB - Intraoral splints are a commonly used dental treatment for a variety of conditions. Because such splints alter the condyle-disc-fossa relationship, they probably change the loading status of the temporomandibular joint (TMJ), including the TMJ disc. Collagen, a major constituent of the disc, acts to resist tensile loading, and it is presumed that the fiber orientations of the individual disc bands reflect their functional loading. Therefore, the purpose of this study was to examine effects of intraoral splint wear on TMJ morphology in general, and collagen orientation of the intra-articular disc in particular. Young adult, female miniature pigs were divided into three groups: open-bite splint, protrusive-bite splint, and unsplinted control. Splints were worn for 2 months, after which the TMJ discs were harvested for histological examination and stereological analysis, and the skulls were cleaned. Although the splints had no effect on skull dimensions, changes were seen in the TMJs. The discs of the protrusively-splinted group showed an increased thickness of the posterior band (P < 0.015) and minor changes in collagen orientation of the anterior band. The most striking change was the presence of a degenerative osseous defect on the medial side of the mandibular condyle in half of the splinted animals. These results indicate that prolonged splint wear can induce remodeling and even injury of TMJ tissues. PMID- 11870600 TI - Experimental confirmation of the sheep model for studying the role of calcified fibrocartilage in hip fractures and tendon attachments. AB - Research has shown that there is a dramatic increase in the fractional area of calcified fibrocartilage from tendon and capsular insertions on the human femoral neck (Vajda and Bloebaum, 1999; Shea et al., 2001b). Additional information regarding the properties of the proximal femur's cortical shell, gained from the use of an animal model, may result in a better understanding of elderly hip fracture since the cortical shell is a significant contributor to the strength of the proximal femur. The objective of the present study was to determine if the greater trochanter's tendon insertions of the human, rat, and sheep differ in terms of morphology and mineralization. The tendons of the greater trochanter of the human, rat, and sheep were observed to insert via a fibrocartilage insertion. The mineral content of the human and sheep calcified fibrocartilage was significantly higher than that of the rat calcified fibrocartilage (P < 0.01). Additionally, the mineral content of the rat cortical bone was significantly higher than that of the human cortical bone (P < 0.01). The mineral content of the calcified fibrocartilage and bone of the human and sheep were not statistically different from each other. There were also more similarities between the bone structure and lacunae density of the human and sheep than between the human and the rat. This suggests that the tendon insertions of the sheep are a better model than the tendon insertions of the rat for the investigation of calcified fibrocartilage in elderly hip fractures. PMID- 11870601 TI - Pulmonary appendix of the short-tailed shrew (Blarina): a unique immunologic organ. AB - The right bronchus of the short-tailed shrew, Blarina brevicauda, terminates in a nonrespiratory pulmonary appendix (PA) containing two bronchial extensions. The experimentally demonstrated ability of these structures to collect and peristaltically expel aspirated material was initially assumed to be a sufficient reason for their developmental persistence, but as bronchus associated lymphoid tissue (BALT) became a subject of immunologic interest in other species, a possible immunologic role for the concentrations of BALT observed in the shrew PA were investigated. As the BALT of the PA contained many well-differentiated plasma cells and numerous particle-containing macrophages, 6-mu paraffin sections were treated with an immunoperoxidase avidin-biotin preparation that chromogenically identified alpha chains of IgA in many of the PA plasma cells and their associated luminal secretions. Also, vascular injections revealed that the PA had a complex relationship with anastomotic sinusoids connecting the bronchial and pulmonary circulation systems, and scanning electron microscopy showed that the luminal epithelial surfaces of the PA were virtually identical to the scattered BALT aggregates in the bronchi of other animals. It thus appeared that these unique structures in the shrew are morphologically and topographically suited to receive aspirated antigens that induce secretory IgA production, while possibly providing other humoral and cellular immunologic products to the general circulation. PMID- 11870602 TI - Successional development of sulfate-reducing bacterial populations and their activities in an activated sludge immobilized agar gel film. AB - A combination of fluorescence in situ hybridization (FISH), microprofiles, and denaturing gradient gel electrophoresis (DGGE) analysis of PCR-amplified 16S rDNA fragments followed by hybridization analysis with specific probes was applied to investigate successional development of sulfate-reducing bacteria (SRB) community structure and in situ sulfide production activity within an activated sludge immobilized agar gel film. In this model biofilm system, since biases arising from biofilm heterogeneity can be ignored, the population dynamics of SRB in the agar gel is directly related to physiological capability and in situ activity of SRB. Microelectrode measurements showed that an anoxic zone was already developed at the beginning (0 day), a first sulfide production of 0.054 mumol H2S m(-2) x s(-1) was detected during the first week, and the rate increased gradually to 0.221 mumol H2S m(-2) x s(-1) in the fifth week. The most active sulfide production zone moved upward to the chemocline and intensified with time to form a narrow zone with high volumetric sulfide production rates. This result coincided with the shift of the spatial distributions of SRB populations determined by FISH. In situ hybridization with probe SRB385 for mainly general SRB of the delta Proteobacteria plus some gram-positive bacteria and probe 660 for Desulfobulbus indicated that the most abundant populations of SRB were primarily restricted to near the oxic/anoxic interface (chemocline). A close observation of the development of the vertical distributions of SRB populations revealed that the cell numbers of Desulfobulbus tripled (from 0.5 x 10(8) to 1.5 x 10(8) cells cm(-3)) near the oxic/anoxic interface. Similar growth (from 1.0 x10(8) to 4.5 x 10(8) cells cm(-3)) of Desulfovibrio-like SRB that hybridized with probe SRB385 was observed. PCR-DGGE followed by hybridization analysis revealed that one Desulfobulbus strain was detected from the beginning, and another strain appeared after 1 week, coinciding with the first detected sulfide production. In addition, three strains hybridizing with probe 687 (possibly Desulfovibrio) were also dominant SRB in the agar gel. PMID- 11870603 TI - Induction of protein aggregation in an early secretory compartment by elevation of expression level. AB - A variety of valuable therapeutic proteins are expressed in mammalian cells. Currently, rate-limiting for secretion of recombinant glycoproteins are activities in the secretory pathway of eukaryotic cells, i.e., folding and glycosylation of the naked polypeptide chain. In this paper we provide evidence that elevation of expression level alone is sufficient to cause intracellular aggregation of a structurally relatively simple glycoprotein, antithrombin III (ATIII). Elevation of expression level by selection for increased drug resistance in Chinese hamster ovary cells stably expressing ATIII resulted in formation of disulfide-bonded aggregates of ATIII. Aggregated ATIII displayed incomplete sialylation and Endo H-sensitivity and located to the endoplasmic reticulum and the cis-Golgi compartment in subcellular fractionations. To explore possible causes for aggregation of ATIII at elevated expression levels we investigated the influence of the two major energy sources of cultured mammalian cells, D-glucose and L-glutamine, on the ATIII-yield. We found that utilization of D-glucose was not limiting for synthesis of ATIII at elevated expression levels. However, the amount of ATIII-synthesized per L-glutamine consumed did not seem to increase steadily with expression level for ATIII, indicating that secretion of ATIII may be limited by the capacity of the cell to utilize L-glutamine. PMID- 11870604 TI - Decreasing the hyphal branching rate of Saccharopolyspora erythraea NRRL 2338 leads to increased resistance to breakage and increased antibiotic production. AB - Mutation and selection for increased resistance to cell-wall synthesis inhibitors led to alterations in the hyphal branching rate of Saccharopolyspora erythraea NRRL 2338. Mutants with decreased branching frequency exhibited increased hyphal strength (estimated by in vitro micromanipulation). As the hyphal strength was increased, this led to a greater proportion of hyphal particles in liquid culture with a hyphal fragment diameter of greater than 88 microm. This, in turn, coincided with proportionately increased antibiotic production. PMID- 11870605 TI - Biokinetics in acidogenesis of highly suspended organic wastewater by adenosine 5' triphosphate analysis. AB - In this paper, we pointed out the problems of using conventional volatile suspended solids (VSS) and chemical oxygen demand (COD) to evaluate biokinetic coefficients, especially for the treatment of highly suspended organic wastewater. We also introduced a novel approach to evaluate biokinetic coefficients by measurement of adenosine 5'-triphosphate (ATP) of microorganisms. The concept of using ATP analysis in biokinetic evaluations with highly suspended wastewater was shown to be effective. This study also showed that the conventional VSS and COD methods were strongly affected by incoming suspended organics in the wastewater and by biokinetics of microorganisms. A cheese processing wastewater was used in evaluating the biokinetics of mesophilic acidogens. The concentration of COD and total suspended solids in the wastewater was 63.3 g/L and 12.4 g/L, respectively. The TSS was 23.6% of total solids concentration. A high ratio of VSS to total suspended solids of 96.7% indicated that most of the suspended particles were organic materials. Lactose and protein were the major organic components contributing COD in the wastewater, and a total of 94.2% of the COD in the wastewater was due to the presence of lactose and protein. Two different physiological conditions where the maximum rates of acetate and butyrate production occurred were tested. These were pH 7 (condition A for acetate production) and pH 7.3 (condition B for butyrate production) at 36.2C, respectively. Based on the molecular structures of the major organic substances and microbial ATP analysis, the residual substrate and microbial concentrations were stoichiometrically converted to substrate COD (SuCOD) and microbial VSS (MVSS), respectively, using correlation coefficients reported previously. These SuCOD and MVSS were simultaneously used to evaluate the biokinetic coefficients using Monod-based mathematical equations. The nonlinear least squares method with 95% confidence interval was used to evaluate biokinetic coefficients. The maximum microbial growth rate, mu(max) and half saturation coefficient, K(s), for conditions A and B were determined to be 9.9 +/- 0.3 and 9.3 +/- 1.0 day(-1) and 134.0 +/- 58.3 and 482.5 +/- 156.5 mg SuCOD/L, respectively. The microbial yield coefficient, Y, and microbial decay rate coefficient, k(d) for conditions A and B were determined to be 0.29 +/- 0.03 and 0.20 +/- 0.05 mg MVSS/mg SuCOD, and 0.14 +/- 0.05 and 0.25 +/- 0.05 day(-1), respectively. Specific substrate utilization rate at condition B was 43.8 +/- 20.6 mg SuCOD/mg MVSS/day, which was 31% higher than that at condition A. PMID- 11870606 TI - Continuous protein recovery from whey using liquid-solid circulating fluidized bed ion-exchange extraction. AB - A liquid-solid circulating fluidized bed (LSCFB) continuous ion-exchange extraction system has been investigated for total protein recovery from whey solutions under various operating conditions. The effectiveness of a dynamic seal was evaluated between the riser and the downcomer, and the best conditions for the establishment of this seal were established. Start-up studies indicated that the system is robust and stable. Under optimal conditions, a productivity of 8.2 g of total protein removed per hour per kilogram of resin was achieved with a protein removal efficiency of 78.4%. However, higher overall protein recovery of up to 90% was also achieved under other conditions, with lower protein concentration in the effluent and a lower overall productivity. PMID- 11870607 TI - Effect of flow regime on the architecture of a Pseudomonas fluorescens biofilm. AB - A comparison of the effects of laminar versus turbulent flow regime on the characteristics of a single-species biofilm is presented. The study was carried out by growing Pseudomonas fluorescens biofilms in a flow cell and studying the different layers of the biological matrix with a confocal laser-scanning microscope. The following conclusions were obtained: i) a higher concentration of cells was found in the upper layers of the microbial films than in their inner layers, regardless of the flow regime; ii) the fraction of cells in the overall biofilm mass decreased with time as the film grew; and iii) under laminar flow the total number of cells was higher than in biofilms formed under turbulent flow, but the latter had a higher number of cells per unit volume. Such conclusions, together with the fact that the biofilms were more dense and stable when formed in contact with turbulent flows, favor the design of more compact and efficient biofilm reactors operating in turbulent conditions. PMID- 11870608 TI - Furfural, 5-hydroxymethyl furfural, and acetoin act as external electron acceptors during anaerobic fermentation of xylose in recombinant Saccharomyces cerevisiae. AB - The electron acceptors acetoin, acetaldehyde, furfural, and 5 hydroxymethylfurfural (HMF) were added to anaerobic batch fermentation of xylose by recombinant, xylose utilising Saccharomyces cerevisiae TMB 3001. The intracellular fluxes during xylose fermentation before and after acetoin addition were calculated with metabolic flux analysis. Acetoin halted xylitol excretion and decreased the flux through the oxidative pentose phosphate pathway. The yield of ethanol increased from 0.62 mol ethanol/mol xylose to 1.35 mol ethanol/mol xylose, and the cell more than doubled its specific ATP production after acetoin addition compared to fermentation of xylose only. This did, however, not result in biomass growth. The xylitol excretion was also decreased by furfural and acetaldehyde but was unchanged by HMF. Thus, furfural present in lignocellulosic hydrolysate can be beneficial for ethanolic fermentation of xylose. Enzymatic analyses showed that the reduction of acetoin and furfural required NADH, whereas the reduction of HMF required NADPH. The enzymatic activity responsible for furfural reduction was considerably higher than for HMF reduction and also in situ furfural conversion was higher than HMF conversion. PMID- 11870609 TI - Characterizing the adhesion of motile and nonmotile Escherichia coli to a glass surface using a parallel-plate flow chamber. AB - A parallel-plate flow chamber was used to measure the attachment and detachment rates of Escherichia coli to a glass surface at various fluid velocities. The effect of flagella on adhesion was investigated by performing experiments with several E. coli strains: AW405 (motile); HCB136 (nonmotile mutant with paralyzed flagella); and HCB137 (nonmotile mutant without flagella). We compared the total attachment rates and the fraction of bacteria retained on the surface to determine how the presence and movement of the flagella influence transport to the surface and adhesion strength in this dynamic system. At the lower fluid velocities, there was no significant difference in the total attachment rates for the three bacterial strains; nonmotile strains settled at a rate that was of the same order of magnitude as the diffusion rate of the motile strain. At the highest fluid velocity, the effect of settling was minimized to better illustrate the importance of motility, and the attachment rates of both nonmotile strains were approximately five times slower than that of the motile bacteria. Thus, different processes controlled the attachment rate depending on the parameter regime in which the experiment was performed. The fractions of motile bacteria retained on the glass surface increased with increasing velocity, whereas the opposite trend was found for the nonmotile strains. This suggests that the rotation of the flagella enables cells to detach from the surface (at the lower fluid velocities) and strengthens adhesion (at higher fluid velocities), whereas nonmotile cells detach as a result of shear. There was no significant difference in the initial attachment rates of the two nonmotile species, which suggests that merely the presence of flagella was not important in this stage of biofilm development. PMID- 11870610 TI - Understanding viral partitioning in two-phase aqueous nonionic micellar systems: 1. Role of attractive interactions between viruses and micelles. AB - The partitioning behavior of viruses in the two-phase aqueous nonionic n-decyl tetra(ethylene oxide) (C10E4) micellar system cannot be fully explained by considering solely the repulsive, steric, excluded-volume interactions that operate between the viruses and the nonionic C10E4 micelles. Specifically, an excluded-volume theory developed recently by our group is not able to quantitatively predict the observed viral partition coefficients, even though this theory is capable of providing reasonable quantitative predictions of protein partition coefficients. To shed light on the discrepancy between the theoretically predicted and the experimentally measured viral partition coefficients, a central assumption underlying the excluded-volume theory that the viruses and the C10E4 micelles interact solely through repulsive, excluded-volume interactions was challenged in this study. In particular, utilizing bacteriophage P22 as a model virus, a competitive inhibition test and a partitioning study of the capsids of bacteriophage P22 were conducted. Based on the results of these two experimental studies, it was concluded that any attractive interactions between the tailspikes of bacteriophage P22 and the C10E4 micelles are negligible. Another experimental study was carried out wherein the partition coefficients of the model viruses, bacteriophages P22 and T4, were measured at various temperatures, and compared with those previously obtained for bacteriophage phiX174. This comparison also indicated that possible attractive, electromagnetic-induced interactions between the bacteriophage particles and the C10E4 micelles cannot be invoked to rationalize the observed discrepancy between the theoretically predicted and the experimentally measured viral partition coefficients. PMID- 11870611 TI - Understanding viral partitioning in two-phase aqueous nonionic micellar systems: 2. Effect of entrained micelle-poor domains. AB - Unlike the partitioning behavior of hydrophilic, water-soluble proteins, the partitioning behavior of viruses in the two-phase aqueous nonionic n-decyl tetra(ethylene oxide) (C10E4) micellar system cannot be fully explained using the excluded-volume theory developed recently by our group. A central assumption underlying the excluded-volume theory--that macroscopic phase separation equilibrium is attained--was therefore challenged experimentally and theoretically. Photographs of the two-phase aqueous C10E4 micellar system were taken for different volume ratios to demonstrate that the entrainment of micelle poor (virus-rich) domains in the macroscopic, top, micelle-rich phase decreases with a decrease in the volume ratio. Partitioning experiments were then conducted with the model virus bacteriophage P22 and the model protein cytochrome c at different operating temperatures for different volume ratios. For bacteriophage P22, the measured viral partition coefficient at each temperature decreased by about an order of magnitude when the volume ratio was decreased from 10 to 0.1, which clearly indicated that entrainment is an important factor influencing viral partitioning. For cytochrome c, the measured protein partition coefficient did not change, which demonstrated that this entrainment effect negligibly influences protein partitioning. A new theoretical description of partitioning was also developed that combines the excluded-volume theory with this entrainment effect. In this theory, one fitted parameter--the volume fraction of entrained micelle poor domains in the macroscopic, top, micelle-rich phase--is used to account for the entrainment. To fit this parameter, only a single partitioning experiment is required for a given volume ratio, irrespectively of the partitioning solute. The new theoretical description of partitioning yielded very good quantitative predictions of the viral partition coefficients. Accordingly, it can be concluded that the primary mechanisms governing viral partitioning in the two-phase aqueous C10E4 micellar system are the entrainment of micelle-poor (virus-rich) domains in the macroscopic, top, micelle-rich phase and the excluded-volume interactions that operate between the viruses and the micelles. PMID- 11870612 TI - Inhibition of sodium butyrate-induced apoptosis in recombinant Chinese hamster ovary cells by constitutively expressing antisense RNA of caspase-3. AB - Sodium butyrate (NaBu) can enhance the expression of genes controlled by some of the mammalian promoters, but it can also inhibit cell growth and induce cellular apoptosis. Thus, the beneficial effect of using a higher concentration of NaBu on a foreign protein expression is compromised by its cytotoxic effect on cell growth. To overcome this cytotoxic effect of NaBu, the expression vector of antisense RNA of caspase-3 was constructed and transfected to recombinant Chinese hamster ovary (rCHO) cells producing a humanized antibody. Using this antisense RNA strategy, rCHO cells (B3) producing a low level of caspase-3 proenzyme were established. When batch cultures of both B3 cells and control cells transfected with antisense RNA-deficient plasmid were performed in the absence of NaBu, both cells showed similar profiles of cell growth and antibody production. Compared with control cell culture, under the condition of 5 mM NaBu addition at the exponential growth phase, expression of antisense RNA of caspase-3 significantly suppressed the NaBu-induced apoptosis of B3 cells and extended culture longevity by >2 days if the culture was terminated at cell viability of 50%. However, compared with control cell culture, the final antibody concentration of B3 cell culture was not increased in the presence of NaBu, which may be due to the loss of cellular metabolic capability resulted from the depolarization of mitochondrial membrane. Taken together, this study suggests that, although expression of antisense RNA of caspase-3 does not improve antibody productivity of rCHO cells, it can suppress NaBu-induced apoptotic cell death of rCHO cells and thereby may reduce problems associated with cellular disintegration. PMID- 11870613 TI - Sol-gel encapsulated enzyme arrays for high-throughput screening of biocatalytic activity. AB - We developed versatile low-cost arrays of sol-gel-encapsulated enzymes (referred to as solzymes) suitable for repeated assays of bioactivity or enzyme inhibition. Sol-gel microstructures containing active enzymes were stabilized on glass at moderate pH and room temperature without harsh calcination. A multi-well bilayer of polydimethylsiloxane was used to support the solzyme array and contain the reaction medium. Each of the 147 microwells has a working volume of 5 muL and contains 50 mug of immobilized enzyme. The solzyme arrays maintained high activity through repeated applications and exhibited superior thermostability compared to soluble enzymes. Among the enzymes used were lipases, glucose oxidase, and horseradish peroxidase. Twenty different lipases and proteases were also used to prepare a hydrolase array, for which bromthymol blue served as a generic indicator of activity. The relative activities of the encapsulated hydrolases correlated closely with those of the soluble hydrolases, illustrating that sol-gel encapsulation preserved the hierarchy of enzyme activity. The development of solzyme arrays paves the way to higher throughput screening of diverse proteins and enzymes, including those that are available only in trace amounts. PMID- 11870614 TI - Different types of V(D)J recombination and end-joining defects in DNA double strand break repair mutant mammalian cells. AB - The end-joining pathway of DNA double-strand break (DSB) repair is necessary for proper V(D)J recombination and repair of DSB caused by ionizing radiation. This DNA repair pathway can either use short stretches of (micro)homology near the DNA ends or use no homology at all (direct end-joining). We designed assays to determine the relative efficiencies of these (sub)pathways of DNA end-joining. In one version, a DNA substrate is linearized in such a way that joining on a particular microhomology creates a novel restriction enzyme recognition site. In the other one, the DSB is made by the RAG1 and RAG2 proteins. After PCR amplification of the junctions, the different end-joining modes can be discriminated by restriction enzyme digestion. We show that inactivation of the 'classic' end-joining factors (Ku80, DNA-PK(CS), ligase IV and XRCC4) results in a dramatic increase of microhomology-directed joining of the linear substrate, but very little decrease in overall joining efficiency. V(D)J recombination, on the other hand, is severely impaired, but also shows a dramatic shift towards microhomology use. Interestingly, two interstrand cross-linker-sensitive cell lines showed decreased microhomology-directed end-joining, but without an effect on V(D)J recombination. These results suggest that direct end-joining and microhomology-directed end-joining constitute genetically distinct DSB repair pathways. PMID- 11870616 TI - Ikaros is critical for B cell differentiation and function. AB - The Ikaros gene encodes a zinc-finger transcription factor required during early B cell development, as B-lineage cells are absent in mice lacking Ikaros. Here we describe a novel Ikaros-targeted mouse line carrying a beta-galactosidase reporter in which low amounts of Ikaros proteins remain expressed. In homozygote animals, B cells are absent during fetal development, but develop postnatally from a reduced pool of precursors. In vitro, the proliferation and differentiation of B-lineage progenitors are severely impaired. These defects are attenuated in vivo, but bone marrow B cells display an unusual pattern of cell surface marker expression and show decreased transcript levels for TdT, Rag-1, Rag-2 and lambda 5. These abnormalities suggest a partial block at the proB cell stage of differentiation. In the periphery, mature B cells exhibit a lower activation threshold but form fewer germinal centers in response to antigenic stimulation. Our results show that Ikaros controls multiple aspects of B cell differentiation and function. PMID- 11870615 TI - Low responsiveness to IFN-gamma, after pretreatment of mouse macrophages with lipopolysaccharides, develops via diverse regulatory pathways. AB - We have investigated the mechanisms by which prior exposure of mouse macrophages to lipopolysaccharides (LPS) induces a state of low responsiveness to subsequent exposure to IFN-gamma. We demonstrate that induction of this state requires both de novo gene expression and the suppression of phosphorylation events that lead to activation of transcription factor Stat1 alpha. These observations are mechanistically consistent with the known induction of suppressors of cytokine signaling (SOCS)-1 and SOCS-3 proteins by LPS. In this regard, we demonstrate that overexpression of either SOCS protein suppresses induction of the mouse inducible nitric oxide synthase (iNOS) gene promoter: apparently by suppressing interactions between Stat1 alpha and IFN-gamma activated sites present in both the iNOS, and interferon regulatory factor-1, gene promoters. The induction of SOCS-1 and SOCS-3 by LPS or IFN-beta (an autocrine/paracrine mediator of LPS induced SOCS-1 mRNA synthesis)occurs by way of multiple protein kinase pathways that include protein tyrosine kinases, protein kinase C, and mitogen-activated protein kinases. These results provide insight that may allow discrimination between LPS-induced inhibition of macrophage functions that are detrimental to the host (e.g. continued exposure to LPS) versus those that might potentially be beneficial (e.g. exposure to subsequent agonists that induce more specific macrophage functions). PMID- 11870618 TI - Uptake and processing of Chlamydia trachomatis by human dendritic cells. AB - Chlamydia trachomatis (CT) causes several sexually transmitted diseases. In 2-5% of cases, CT infection leads to the development of reactive arthritis. Dendritic cells (DC) are central in T cell priming and the induction of antigen specific immunity. Here we have studied the uptake and processing of CT serovar L2 by human DC, and their ability to present CT antigens to both CD4(+) and CD8(+) T cells. We show that the entry of CT was mediated by the attachment of CT to heparan sulfates and could be inhibited by heparin. There was no inhibition of uptake by an agent which blocks micropinocytosis. Infecting DC with CT led to their activation and the production of IL-12 and TNF-alpha but not IL-10. Following invasion, CT was confined to distinct vacuoles which were visualized with anti-CT antibodies using confocal microscopy. Unlike with epithelial cells, these vacuoles did not develop into characteristic inclusion bodies. In the first 48 h, CT(+) vacuoles were negative for Lamp-1 and MHC class II. Despite no obvious co-localization between CT vacuoles and MHC loading compartments, infected DC efficiently presented CT antigens to CD4(+) T cells. Infected DC also expanded CT specific CD8(+) T cells, allowing us to generate a number of CT reactive CD8(+) T cell clones. There is still controversy about the importance of chlamydia-specific CD8(+) T cell responses in patients with arthritis. This is largely due to the difficulties in studying CTL responses at the clonal level. The use of DC as antigen-presenting cells should enable more detailed characterization of these CTL responses. PMID- 11870617 TI - In vivo activation of melanoma-specific CD8(+) T cells by endogenous tumor antigen and peptide vaccines. A comparison to virus-specific T cells. AB - Many new types of vaccines against infectious or malignant diseases are currently being proposed. Careful characterization of the induced immune response is required in assessing their efficiency. While in most studies human tumor antigen specific T cells are analyzed after in vitro re-stimulation, we investigated these T cells directly ex vivo using fluorescent tetramers. In peripheral blood lymphocytes from untreated melanoma patients with advanced disease, a fraction of tumor antigen (Melan-A/MART-1)-specific T cells were non-naive, thus revealing tumor-driven immune activation. After immunotherapy with synthetic peptides plus adjuvant, we detected tumor antigen-specific T cells that proliferated and differentiated to memory cells in vivo in some melanoma patients. However, these cells did not present the features of effector cells as found in cytomegalovirus specific T cells analyzed in parallel. Thus, peptide plus adjuvant vaccines can lead to activation and expansion of antigen specific CD8(+) T cells in PBL. Differentiation to protective CD8(+) effector cells may, however, require additional vaccine components that stimulate T cells more efficiently, a major challenge for the development of future immunotherapy. PMID- 11870619 TI - Generation of adult-like antibody avidity profiles after early-life immunization with protein vaccines. AB - The capacity to induce high-avidity antibodies following early-life immunization has long been questioned, and the possibility of inducing such antibodies soon after birth is a recognized goal for a number of vaccination strategies. Therefore, we assessed the capacity to develop high-avidity antibodies to peptidic vaccines in 1-week-old BALB/c mice. The dynamics of the generation of antibody molecules of increasing avidity were analyzed on circulating serum antibodies and, where feasible, at the single-cell level on spleen and bone marrow antibody-secreting cells. Two alum-adsorbed protein-based human vaccines, tetanus toxoid (TT) and pertussis toxin, induced neonatal antibody responses with adult-like avidity profiles. This was confirmed at the level of spleen and bone marrow ASC. In contrast, immunization with TT-P30, a 21-mer synthetic peptide containing a TT-immunodominant epitope, trinitrophenyl hapten (TNP) conjugated to ovalbumin or TNP conjugated to Ficoll, induced a much lower avidity profile in early life than in adults. These observations indicate that in murine models the avidity maturation of T cell-dependent antibody responses induced by structurally complex protein vaccines can be fully elicited after early life immunization, as opposed to the maturation of responses induced with short peptides or hapten based vaccines. PMID- 11870620 TI - Expression and role of CD14 in mice sensitized to lipopolysaccharide by Propionibacterium acnes. AB - Propionibacterium acnes-primed mice develop an IFN-gamma-dependent hypersensitivity towards LPS. Since CD14 plays a key role in LPS-induced cell activation the regulation and function of CD14 in this sensitization process were studied in IFN-gamma R-/- and the respective wild-type (wt) mice. In unprimed mice, CD14 (mRNA and protein) was either absent (liver) or only weakly expressed in organs (spleen, lung) and in plasma. Priming with P. acnes led to a moderate, mainly IFN-gamma-dependent up-regulation of CD14. LPS challenge of unprimed mice induced an IFN-gamma-independent increase in CD14 mRNA and CD14 protein. LPS challenge of P. acnes-primed mice induced a strong CD14 overexpression. This response was completely absent in IFN-gamma R-/- mice and is therefore strictly IFN-gamma-dependent. The requirement for CD14 in LPS hyper-responsiveness was assessed by comparing CD14-/- and the respective wt mice with respect to their ability to produce TNF and IFN-gamma, two recognized indices of LPS activity. LPS challenge without priming led to a weaker cytokine reaction in CD14-/- than in wt mice. However, priming with P. acnes enhanced the cytokine response to LPS in both wt and CD14-/- mice, although in the latter absolute levels of cytokines were lower. Therefore, hyperreactivity to LPS is characterized by an up regulation of CD14, but the sensitization by P. acnes is not CD14 dependent. PMID- 11870621 TI - A role for the src family kinase Fyn in NK cell activation and the formation of the repertoire of Ly49 receptors. AB - NK cell function is regulated by a dual receptor system, which integrates signals from triggering receptors and MHC class I-specific inhibitory receptors. We show here that the src family kinase Fyn is required for efficient, NK cell-mediated lysis of target cells, which lack both self-MHC class I molecules and ligands for NKG2D, an activating NK cell receptor. In contrast, NK cell inhibition by the MHC class I-specific receptor Ly49A was independent of Fyn, suggesting that Fyn is specifically required for NK cell activation via non-MHC receptor(s). Compared to wild type, significantly fewer Fyn-deficient NK cells expressed the inhibitory Ly49A receptor. The presence of a transgenic Ly49A receptor together with its H 2(d) ligand strongly reduced the usage of endogenous Ly49 receptors in Fyn deficient mice. These data suggest a model in which the repertoire of inhibitory Ly49 receptors is formed under the influenced of Fyn-dependent NK cell activation as well as the respective MHC class I environment. NK cells may acquire Ly49 receptors until they generate sufficient inhibitory signals to balance their activation levels. Such a process would ensure the induction of NK cell self tolerance. PMID- 11870622 TI - The membrane attack complex of complement induces caspase activation and apoptosis. AB - Activation of the terminal pathway of the complement system leads to insertion of terminal complement complexes (C5b-9) into the cell membrane, which may induce cytolysis. Recent data indicate that the terminal complement pathway can also result in apoptosis in vivo. To further define the cell death pathway induced by complement, we examined induction of apoptosis by complement in vitro. Rat mesangial cells opsonized with a complement-activating antibody and exposed to rat serum as a complement source underwent apoptotic cell death in a time- and dose-dependent fashion, as demonstrated by membrane exposure of phosphatidylserine and fragmentation of nuclei. No significant apoptosis was detected in either cultures treated with C6-deficient serum or in control cultures. The pan-caspase-inhibitor zVAD-fmk inhibited complement-induced apoptosis completely. In line with this observation, complement induced cleavage and activation of caspase 3. Importantly, cellular exposure to purified cytolytically inactive C5b-9, in the absence of antibody and early complement components, also resulted into caspase activation and apoptosis. Together, these results indicate that C5b-9 is involved in induction of apoptosis via a caspase dependent pathway. Apoptosis as a consequence of complement-mediated cell damage may provide an explanation for the presence of apoptosis in inflammatory processes, for instance in hyperacute xenograft rejection. PMID- 11870623 TI - Perforin- and Fas-dependent mechanisms of natural killer cell-mediated rejection of incompatible bone marrow cell grafts. AB - Natural killer (NK) cells eliminate target cells infected with intracellular pathogens and tumor cells by employing the granule exocytosis and death receptor pathways. They also mediate the acute rejection of incompatible bone marrow cell (BMC) grafts. However, the cytotoxic mechanisms employed during acute BMC graft rejection are obscure. Throughout these studies, BMC graft rejection was compared between two inbred strains of mice: 129 mice, which apparently use perforin- and Fas-dependent cytotoxicity, and C57BL/6 (B6) mice, which are able to exploit perforin- and/or Fas-independent mechanisms. Using perforin-knockout (PKO) mice, we have determined that the granule exocytosis pathway can play a major role in NK cell-mediated rejection of allogeneic and MHC class I-deficient BMC, depending upon the genetic background of the recipient and the environmental housing conditions. Although the granule exocytosis pathway seems to be the most potent cytolytic mechanism of NK cell-mediated rejection, alternative perforin independent mechanisms, such as death receptor-induced apoptosis, also exist. By preventing both perforin- and Fas-mediated interactions concurrently, we observed that 129 mice were impaired in mediating MHC class I-deficient BMC rejection, while B6 mice maintained strong rejection capacities. The administration of neutralizing TNF antibodies to B6PKO mice before challenging with Fas and MHC class I double-deficient BMC still did not reverse rejection. Thus, our studies reveal the relative importance of perforin-, Fas-, and TNF-based cytotoxicity in NK cell-mediated rejection of incompatible BMC. PMID- 11870624 TI - Bag-1 up-regulation in anti-CD4 mAb treated allo-activated T cells confers resistance to apoptosis. AB - The non-depleting anti-CD4 mAb RIB5/2 is a powerful inducer of tolerance to MHC incompatible renal and heart allografts in rat recipients. In vitro the mAb blocks the proliferation and cytokine production of alloreactive T cells. To learn more about the mechanism of anti-CD4-mediated suppression, we applied differential display reverse transcription-PCR to identify differences at mRNA level between T cells stimulated by alloantigen in the presence or absence of anti-CD4 mAb. A sequence alignment of a 550-bp DNA fragment appearing only in anti-CD4 mAb-treated cells resulted in at least 95% homology to a mouse cDNA encoding for the anti-apoptotic protein Bag-1. Further investigation of Bag-1 expression during mixed lymphocyte reactions revealed a three- to fourfold up regulation of Bag-1 mRNA expression in anti-CD4 mAb-treated allogeneic cultures which was confirmed at protein level. Bag-1 up-regulation was associated with an increase resistance to apoptosis of T cells from anti-CD4 mAb-treated cultures. Application of antisense oligonucleotides specific for Bag-1 reduced Bag-1 protein expression and restored susceptibility to apoptosis. In addition, up regulation of Bag-1 mRNA could also be detected in graft-infiltrating T cells from anti-CD4 mAb-treated rats in vivo. Thus, the expression of Bag-1 in a subset of anti-CD4 mAb-treated alloreactive T cells conferred resistance against apoptosis, potentially contributing to the long-term survival of these cells. PMID- 11870625 TI - Evolution of NK receptors: a single Ly49 and multiple KIR genes in the cow. AB - Natural killer (NK) cell receptors for classical MHC class I molecules are encoded by the killer Ig-like receptor (KIR) multigene family in humans and other primates. Mouse NK cells, however, employ a completely different multigene family, the C-type lectin-like Ly49 genes, to perform the same function. This example of functional convergent evolution raises the question of what type of receptors are found in non-primate and non-rodent mammals. By screening a bovine spleen cDNA library, we isolated an Ly49 gene from the cow (Bos Taurus) and show by genomic Southern blotting that it is likely a single copy gene in this species. The coding region is intact and has an immunoreceptor tyrosine-based inhibition motif (ITIM) in the cytoplasmic domain, suggesting a role as an inhibitory receptor. We have also identified several bovine cDNA clones related to KIR and show that at least one has an intact open reading frame with two ITIM. Evidence for multiple KIR-like genes in the cow was obtained by Southern blotting and we found that at least two of these genes contain an ancient retro-element present in all human KIR genes. These results suggest that the cow and primate KIRgene families arose from a common ancestral gene but amplified independently. Furthermore, these findings indicate that the existence of multiple Ly49 genes may be a phenomenon unique to rodents. PMID- 11870626 TI - Cross-presentation of virus-like particles by skin-derived CD8(-) dendritic cells: a dispensable role for TAP. AB - Virus-like particles (VLP) induce efficient CTL responses although they do not carry any genetic information. Here, we analyzed MHC class I associated presentation of VLP-derived CTL-epitopes in vivo. After intradermal injection of VLP containing the immunodominant epitope (p33) of lymphocytic choriomeningitis virus (p33-VLP), presentation of peptide p33 in draining lymph nodes was largely restricted to CD8(-) skin-derived dendritic cells (DC). Surprisingly, and in contrast to findings with tumor cells, TAP1-deficient DC and macrophages mediated efficient cross-presentation of VLP-derived p33 in vivo and in vitro. However, the ability of TAP1-deficient DC to cross-present p33-VLP was reduced compared to wild-type DC, indicating that in DC, both TAP-dependent and TAP-independent pathways were operative. In contrast, macrophages cross-presented p33-VLP normally in the absence of TAP. The TAP-dependent pathway of cross-presentation is therefore confined to DC while both macrophages and DC harbor the TAP independent pathway. In summary, the results show that VLP-derived epitopes are cross-presented by CD8(-) DC in vivo in a partial TAP-independent fashion and highlight important differences in the processing machinery of DC versus macrophages. PMID- 11870627 TI - Cellular and humoral immune responses to a human pancreatic cancer antigen, coactosin-like protein, originally defined by the SEREX method. AB - Among a number of human tumor antigens identified using the serological analysis of recombinant cDNA expression libraries (SEREX), only MAGE-1, tyrosinase, and NY ESO-1 have been reported to be immunogenic tumor antigens that have the potential to elicit both humoral and cellular immunity. In this study, we determined whether our SEREX-defined pancreatic cancer antigens could be recognized by CTL, and report that one SEREX-defined antigen, coactosin-like protein (CLP), encoded cellular epitopes recognized by HLA-A2-restricted and tumor-reactive CTL. Three CLP peptides at positions 15-24, 57-65, and 10-113 possessed the ability to induce HLA-A2-restricted and tumor-reactive CTL from the PBMC of cancer patients. Subsequently, humoral responses to these peptides were investigated. IgG antibodies specific to the CLP 15-24, 57-65, and 104-113 peptides were detected in sera from 12, 0, and 12 of 12 cancer patients tested, and were also found in 5, 0, and 0 of 9 healthy donors, respectively. IgE antibodies specific to these peptides were also detected in sera from certain cancer patients and healthy donors. Since peptide-specific IgE was detected, type-I allergy to these peptides was tested. Unexpectedly the CLP 57-65 peptide, to which IgE was found in only 2 healthy donors, but not the other two peptides, was found to elicit an immediate type hypersensitivity in all 10 healthy volunteers tested. These results indicate that identical antigenic peptides can be recognized by both cellular and humoral immune systems to a tumor-associated antigen. The CLP 15-24 and 104-113 peptides might be appropriate vaccine candidates for peptide-based immunotherapy of HLA A2(+) cancer patients. PMID- 11870628 TI - Activation of pertussis toxin-sensitive CXCL12 (SDF-1) receptors mediates transendothelial migration of T lymphocytes across lymph node high endothelial cells. AB - In this study we examined the role of chemokines in regulating T lymphocyte transmigration across the lining high endothelial cells (HEC) of high endothelial venules (HEV). The roles played by CCL21 (SLC), CCL19 (MIP-3 beta, ELC) and CXCL12 (SDF-1) were assessed using an in vitro transendothelial migration culture system, which constitutively supports high levels of lymphocyte transmigration. We determined that transmigration of T lymphocytes across HEC is inhibitable by treatment of the T lymphocytes with pertussis toxin (PTX) (80% inhibition). This was attributed to blockade of Gi-protein coupled receptors of T lymphocytes, since a non-ADP-ribosylating form of PTX had no significant effect on transendothelial migration. Inhibition of Gi-protein-coupled receptors on the endothelium had no effect on T cell transmigration. Treatment of T lymphocytes with a desensitizing concentration of CXCL12 caused a 60% reduction in T lymphocyte migration across HEC, and the CXCR4 antagonist SDF-1P2G reduced transmigration by 40%. Desensitizing concentrations of CCL21 and CCL19 had no significant effects on T lymphocyte transendothelial migration. Homologous desensitization of T lymphocytes to each chemokine was confirmed in a transwell migration assay. An approximately 3-kb mRNA corresponding to rat SDF-1 beta was constitutively expressed in HEC and cell surface CXCL12 was detectable by enzyme linked immunosorbent assay. Together, these findings support a pivotal role for HEC-expressed CXCL12 and its receptor on T cells in the regulation of T lymphocyte homing to lymph nodes. PMID- 11870629 TI - CD1d on antigen-transporting APC and splenic marginal zone B cells promotes NKT cell-dependent tolerance. AB - CD1d-reactive NKT cells are required for the development of antigen-specific T regulatory (Tr) cells responsible for mediating systemic tolerance induced through an immune privileged site such as the eye. The aim of this study was to elucidate the cellular source of CD1d needed for NKT cell activation. Transforming growth factor beta (TGF beta)-2-treated peritoneal exudate cells (PEC) functionally resemble "eye-derived" antigen-presenting cells (APC) and contribute to the generation of Tr cells both in vitro and in vivo. However, when TGF beta-2-treated PEC were pretreated with CD1d-specific antibodies or lacked CD1d expression they failed to induce Tr cells. In addition, we show that the subpopulation of marginal zone (MZ) B cells within the spleen is necessary for induction of Tr cells and that they also must express CD1d. The role of MZ B in tolerance is novel and previously unexplored. Thus, CD1d is necessarily expressed on putative eye-derived APC and splenic MZ B cells for efficient generation of Tr cells in CD1d-reactive NKT cell-dependent tolerance. PMID- 11870630 TI - Infection with Brugia microfilariae induces apoptosis of CD4(+) T lymphocytes: a mechanism of immune unresponsiveness in filariasis. AB - In humans infected with lymphatic filariasis, microfilaraemia [the presence of microfilariae (Mf) in the blood] is generally associated with both poor antigen (Ag)-specific proliferative responses and with protection from severe disease. Clonal deletion has been suggested as one possible mechanism by which parasite reactive lymphocytes, that may be capable of mediating resistance and/or immunopathology, are silenced in asymptomatic carriers. In this study we demonstrate that splenic lymphocytes from mice infected with microfilariae of Brugia pahangi display an Ag-specific proliferative defect. However, these cells were not completely unresponsive since they produced high levels of Ag-specific IFN-gamma. Using TdT-mediated dUTP-biotin nick end labeling for flow cytometry, CD4(+) lymphocytes from Mf-infected mice cultured with Ag showed high levels of apoptosis when compared to those from L3-infected mice which proliferated well in response to Ag. Treatment of Ag-stimulated cultures with aminoguanidine (AMG), an inhibitor of inducible nitric oxide synthase, rescued the CD4(+) T cells from apoptosis and reversed the proliferative defect. Furthermore, carboxyfluorescein diacetate succinimidyl ester labeling allowed the visualization of dividing CD4(+) T cells in cultures from Mf-infected animals only in the presence of AMG. We hypothesize that CD4(+) T cells indirectly trigger their own apoptosis by secreting significant quantities of IFN-gamma resulting in the induction of high levels of nitric oxide, and the subsequent elimination of effector T cells. Our findings are the first direct evidence that infection with Brugia Mf can selectively induce lymphocyte apoptosis, a phenomenon that could contribute to the proliferative defect and parasite persistence associated with the microfilaraemic state in the infected human. PMID- 11870631 TI - NK cells developing in vitro from fetal mouse progenitors express at least one member of the Ly49 family that is acquired in a time-dependent and stochastic manner independently of CD94 and NKG2. AB - NK cells developing in vitro from fetal progenitors in the presence of IL-2 are phenotypically and functionally indistinguishable from mature adult NK cells, with the exception that they generally lack surface expression of any of the Ly49 molecules that have previously been examined. Using two recently developed anti Ly49 mAb, we show here that most of these NK cells in fact express high levels of at least one previously uncharacterized member of the Ly49 family, most likely Ly49E. Detailed kinetic and clonal analysis revealed that these Ly49 molecules were acquired in a progressive and stochastic manner independently of CD94 and NKG2. CD94 and NKG2 were both expressed early in NK cell development, sometimes in the absence of NK1.1, with CD94 invariably being expressed at two different levels. IL-4 differentially inhibited the expression of CD94 and Ly49 receptors, but had little or no effect on the expression of NKRP1 molecules. PMID- 11870632 TI - Human NK1 and NK2 subsets determined by purification of IFN-gamma-secreting and IFN-gamma-nonsecreting NK cells. AB - The in vivo existence of human NK cell subsets similar to Th1 and Th2 cells was demonstrated in freshly isolated IFN-gamma-secreting and IFN-gamma-nonsecreting NK cells. The IFN-gamma-secreting NK subset showed a typical cytokine pattern with predominant expression of IFN-gamma, but almost no IL-4, IL-5 and IL-13. In contrast, the IFN-gamma-nonsecreting NK subset was composed of IL-4, IL-5 and IL 13-producing NK cells. Short-time stimulation or 2 weeks of in vitro differentiation of NK cells led to distinct patterns of cytokine production similar to freshly-purified IFN-gamma (+) or IFN-gamma (-) NK cell subsets. NK cells stimulated with IL-12 produced increased levels of IFN-gamma and decreased levels of IL-4. In contrast, stimulation of NK cells with IL-4 inhibited IFN gamma, but increased IL-13 production. Freshly-purified IFN-gamma (+) and IFN gamma (-) or in vitro differentiated NK1 and NK2 subsets showed similar cytotoxicity to K562 cells. These results demonstrate that circulating NK cells retain effector subsets in humans with distinct cytokine profiles and may display different inflammatory properties. PMID- 11870634 TI - The selective triggering of CD40 on keratinocytes in vivo enhances cell-mediated immunity. AB - CD40 is expressed on a wide array of hematopoietic and non-hematopoietic cells, including keratinocytes. The pivotal in vivo function of CD40 on hematopoietic cells in the regulation of both humoral and cell-mediated immunity is well established. However, whether CD40 expression on non-hematopoietic cells influences immunity has until now not been addressed. Therefore, we transgenically expressed human CD40 (hCD40) under the control of the keratin 14 promoter to drive expression of hCD40 in basal keratinocytes of mice. When we selectively engaged hCD40 in vivo on the keratinocytes of these mice, the keratinocytes secreted TNF-alpha, resulting in dendritic cell migration to draining lymph nodes and enhanced in vitro T cell priming to an epicutaneously applied chemical sensitizer. Exclusive CD40 engagement on keratinocytes during a contact hypersensitivity response displayed exacerbated and prolonged cutaneous immune reactions relative to control mice. Thus, CD40 engagement on non hematopoietic cells, such as keratinocytes, can amplify cutaneous and regional T cell responses in vivo. PMID- 11870633 TI - Enhanced T cell responses contribute to the genetic predisposition of CD8 mediated spontaneous autoimmunity. AB - A number of factors have been demonstrated to influence the induction of pathogenic autoimmune responses, including the loss of regulatory T cells. To assess the contribution of regulatory T cells in CD8(+) T cell-mediated autoimmunity, RIP-gp/P14 double-transgenic mice expressing the lymphocytic choriomeningitis virus (LCMV) glycoprotein (gp) on pancreatic beta-islet cells, together with T cells expressing an LCMV-gp-specific T cell receptor (TCR), were crossed to RAG 2-deficient mice. The loss of potentially regulatory T cells, however, did not contribute to diabetes induction. Surprisingly, both RIP-gp/P14 RAG(+/-) and RIP-gp/P14-RAG(-/-) developed spontaneous disease, suggesting an influence of the 129 genetic background on disease susceptibility. Further studies demonstrated that disease susceptibility was not due to nonspecific T cell activation, nor to enhanced cross-presentation of LCMV-gp, nor to decreased expression levels of the negative regulatory molecule CD5. Disease susceptibility did associate, however, with enhanced T cell responses. Thus, T cell hyperactivity combined with various genetic factors may predispose an individual to autoimmunity. PMID- 11870635 TI - Salivary gland B cell lymphoproliferative disorders in Sjogren's syndrome present a restricted use of antigen receptor gene segments similar to those used by hepatitis C virus-associated non-Hodgkins's lymphomas. AB - Sjogren's syndrome (SS) represents a pathological model of the evolution from polyclonal B lymphocyte activation to oligoclonal/monoclonal B cell expansion, which may culminate in the development of a malignant lymphoproliferative disease. The different phases of this process are usually marked by the appearance of antigen-driven activated B cell clones, which are commonly IgM positive and with rheumatoid factor (RF) activity. However, the agent(s) able to trigger B cell proliferation is still unknown. A similar pathogenetic mechanism exist in mixed cryoglobulinemia, another autoimmune disease that often evolves to non-Hodgkin's lymphoma (NHL) and in which hepatitis C virus (HCV) infection has been demonstrated to play an etiopathogenetic role. In the present study, we cloned and sequenced the antigen receptor (IgR) variable region genes of SS associated monoclonal non-neoplastic lymphoproliferations and compared them with those of our previous reported HCV-associated NHL, to derive clues on the antigen(s) that sustains SS. The results obtained showed remarkable homologies between the antigen combinatory regions of the IgR expressed by both diseases. These homologies concern: a) the specific combinations of heavy and light variable region genes; b) the limited length of complementarity-determining regions (CDR3); c) the homology with antibodies with RF activity; d)the amino acid sequences of CDR3 in which common somatic mutations are present that possibly determine the antigen-binding specificity. In conclusion, although there are significant differences between SS and HCV-associated lymphoproliferative diseases, they share many molecular characteristics, which suggest an immunological cross-reactivity or molecular mimicry among the agents that underlie these disorders. PMID- 11870636 TI - The pT alpha promoter and enhancer are direct targets for transactivation by E box-binding proteins. AB - The pre-T alpha (pT alpha) gene encodes a protein that forms the pre-TCR together with the newly expressed TCR beta chain during a defined stage of early T cell development. The restricted expression of this gene has been suggested to depend on a promoter and an enhancer element containing E boxes, which are potential binding sites for basic helix-loop-helix (bHLH) proteins. We here report that increased expression of E47 and E12 mRNA in CD4(-)CD8(-)TCR(-) thymocytes correlates with the initiation of pT alpha transcription. Furthermore, electrophoretic mobility shift assays suggested that E47 binds directly to both the pT alpha promoter and enhancer in vitro. Ectopic expression of E47 in non lymphoid HeLa cells resulted in a dose- and template-dependent activation of these control elements. In addition to this, ectopic expression of E47 in combination with the ets-protein Pu.1 and/or the Pu.1-interacting protein(Pip-1), resulted in a synergistic effect on the activity of the pT alpha control elements. Thus, we suggest that E47, Pu.1 and Pip-1 are directly involved in the regulation of pT alpha expression in early T cell development. PMID- 11870639 TI - In vitro behavior of sintered powder injection molded Ti-6Al-4V/HA. AB - This article reports the morphology and mechanical properties of sintered powder injection molded Ti-6Al-4V/HA parts in a simulated physiological environment. Sintered Ti-6Al-4V/HA parts were immersed in a simulated body fluid (SBF) with ion concentrations that were comparable to those of human blood plasma for a total period of 12 weeks. At intervals of 2 weeks, the immersed Ti-6Al-4V/HA parts were analyzed with the use of scanning electron microscopy (SEM), X-ray diffractometry (XRD), and inductively coupled plasma-atomic emission spectroscopy (ICP-AES). Mechanical properties such as flexural strength, flexural modulus, compressive strength, and compressive modulus were also evaluated. Results showed that complete dissolution of the more soluble phases such as tricalcium phosphate (TCP), tetracalcium phosphate (TTCP), and calcium oxide (CaO) were found after 2 weeks of immersion in SBF. ICP analysis showed that high calcium concentration release of around 200 ppm was observed in the SBF solution after 2-4 weeks of immersion, indicating that dissolution has taken place. Next, a gradual decrease in calcium concentration release in the SBF solution was observed after immersion for 4-6 weeks, with increasing amounts of calcium phosphate precipitates being observed on the Ti-6Al-4V/HA surface. Mechanical properties such as strength and modulus were found to deteriorate during 2-4 weeks of immersion, followed by gradual increment as the immersion period increased. This study also showed that parts sintered at 1150 C exhibited faster dissolution and precipitation rates than parts sintered at 1050 C in a physiological environment. PMID- 11870640 TI - Characterization of new acrylic bone cements prepared with oleic acid derivatives. AB - Acrylic bone-cement formulations were prepared with the use of a new tertiary aromatic amine derived from oleic acid, and also by incorporating an acrylic monomer derived from the same acid with the aim of reducing the leaching of toxic residuals and improving mechanical properties. 4-N,N dimethylaminobenzyl oleate (DMAO) was used as an activator in the benzoyl-peroxide radical cold curing of polymethyl methacrylate. Cements that contained DMAO exhibited much lower polymerization exotherm values, ranging between 55 and 62 C, with a setting time around 16--17 min, depending on the amine/BPO molar ratio of the formulation. On curing a commercial bone cement, Palacosreg R with DMAO, a decrease of 20 C in peak temperature and an increase in setting time of 7 min were obtained, the curing parameters remaining well within limits permitted by the standards. In a second stage, partial substitution of MMA by oleyloxyethyl methacrylate (OMA) in the acrylic formulations was performed, the polymerization being initiated with the DMAO/BPO redox system. These formulations exhibited longer setting times and lower peak temperatures with respect to those based on PMMA. The glass transition temperature of the experimental cements were lower than that of PMMA cement because of the presence of long aliphatic chains of both activator and monomer in the cement matrix. Number average molecular weights of the cured cements were in the range of 1.2x10(5). PMMA cements cured with DMAO/BPO revealed a significant (p<0.001) increase in the strain to failure and a significant (p<0.001) decrease in Young's modulus in comparison to Palacosreg R, whereas ultimate tensile strength remained unchanged. When the monomer OMA was incorporated, low concentrations of OMA provided a significant increase in tensile strength and elastic modulus without impairing the strain to failure. The results demonstrate that the experimental cements based on DMAO and OMA have excellent promise for use as orthopaedic and/or dental grouting materials. PMID- 11870641 TI - Deposition of (90)YPO(4) and (144)CePO(4) radioisotopes on polymer surfaces for radiation delivery devices. AB - Intravascular irradiation with beta emitters inhibits restenosis in arteries after balloon angioplasty or stent implantation. Yttrium-90 ((90)Y, T(1/2)=64 h) and cerium-144 ((144)Ce, T(1/2)=286 d) emit beta particles (E(max)=2.28--3.50 MeV) having an ideal energy range for brachytherapy delivery system. In this article, a previously reported method for depositing (32)P on poly(ethylene terephtalate) (PET) surfaces is generalized and modifications that allow deposition of other beta-emitting radioisotopes, such as (90)Y and (144)Ce, are demonstrated. PET films were first coated with chitosan hydrogel and then adsorbed different amounts of phosphoric acid (PA) in aqueous solutions. Yttrium was deposited onto the surface as YPO(4) after the films were immersed in YCl(3) solutions. 1 muCi (90)YCl(3) (2 x 10(-9) g) was used in each sample as a tracer for measuring the deposition efficiency, which is defined as the percentage of YCl(3) deposited on the surface compared to the amount of YCl(3) in solutions before the deposition. In order to improve the safety of brachytherapy treatments, polyurethanes were used to seal the deposited radioisotopes on the surface to minimize the leakage of the isotopes into the patients. The generality of this method presented here for a wide variety of particular radioisotopic components allows design of a broad range of versatile radioisotope sources. PMID- 11870642 TI - The contribution of coating microstructure to degradation and particle release in hydroxyapatite coated prostheses. AB - Plasma-sprayed coatings of hydroxyapatite powder are widely used on hip replacements. Commercially, they are supplied by a large number of companies and thus offer different coating design philosophies. This study focuses on a retrieved prosthetic stem that exhibited coating loss on the femoral stem occurring concurrently with third-body wear. The purpose of the research was to establish possible links between the coating microstructure and the clinical findings. A coated stem and cup were sectioned and the cross section was prepared to reveal the coating microstructure. Characterization included X-ray diffraction, FTIR spectroscopy, and crystalline particle quantification within the coating. It was found that the coating has a high amorphous content that provides fast resorption. The amount of crystalline particles increased on the distal location of the stem, the threads of the acetabular shell, and was generally higher on the cup. Accelerated degradation illustrated how the coating may be a particle-generating source by preferential dissolution of the amorphous phase, possibly allowing liberation of crystalline areas and other particulates at the substrate-coating interface. Such particles mainly include the less soluble hydroxyapatide formed from unmelted particles in the plasma or recrystallisation in the coating, but may also include entrapped grit lodged in the substrate during the roughening process. This study accents the importance of coating microstructure in understanding coating resorption. PMID- 11870643 TI - Degradation characteristics of alpha and beta tri-calcium-phosphate (TCP) in minipigs. AB - In seven Goettingen minipigs 3.5--4.7-ml cancellous bone defects were created in the area of the tibial head on both sides. The defects were filled with alpha-TCP or beta-TCP (tricalciumphosphate). ITI implants (Straumann, Freiburg, Germany) of 3.2 x 12-mm length were inserted into the underlying ceramic substitutes. Two additional pigs were used as control. Within the periods of observation (4, 16, 20, 28, 46, 68, and 86 weeks) fluorescent dyes were applied. Nondecalcified thin sliced sections were examined by means of light and fluorescence microscopy. In addition microangiography and microradiography were performed. Bony regeneration occurred basally and on the sides of the defect according the angiogenetic reossification pattern. Resorption was due to a hydrolytic and cellular degradation process. After 46 weeks histomorphological evaluation showed an incomplete osseointegration of the simultaneously implanted dental implants. The bone contact surface ratio was lower than 25%. After 86 weeks 95--97% of both alpha- and beta-TCP were resorbed. Ceramic residuals stayed within the newly formed trabeculae thus resisting further degradation until remodeling occurred. Both alpha- and beta-TCP show a comparable degradation process. At the 86-week postoperative point only small residuals of the ceramic can be found. These residuals stay within the newly formed trabeculae, which show a functional orientation. In comparison control defects showed only sparse reossification. The beta-TCP material shows an accelerated degradation mode and has an optimal reactivity with the surrounding tissues. According to the results of this animal experiment both materials can be classified as bone-rebuilding materials. PMID- 11870644 TI - Surface modification of an experimental silicone rubber aimed at reducing initial candidal adhesion. AB - Silicone rubber, which is a widely used biomaterial, is often used to make soft liners for permanent denture. Colonization of denture soft lining materials by Candida albicans can result in clinical problems. The aim of this study was to chemically modify the surface of an experimental silicone rubber in order to produce a silicone that was less susceptible to candidal colonization. Surface modification was carried out with the use of argon-plasma bombardment followed by silane treatment, which caused the incorporation of either hydrophilic or hydrophobic functional groups onto the surface. Changes in water contact angles and chemical analysis of the materials with scanning ion mass spectroscopy confirmed surface changes. In vitro assays were carried out using C. albicans to measure levels of adherence to the surface-modified silicone after 1 h. C. albicans exhibited very low adherence to all silane-treated surfaces, whether hydrophobic or hydrophilic. This led to the conclusion that incorporated long chain functional groups were inhibiting the adherence of the yeast, possibly by the formation of a barrier between the surface of the material and the yeast. In conclusion, silane surface treatment of an experimental silicone rubber has been successful in reducing candidal adherence. PMID- 11870645 TI - Functional and morphological evaluation of a low-weight, monofilament polypropylene mesh for hernia repair. AB - With more than 1 million implantations per year worldwide, mesh repair has become a standard procedure for the treatment of hernias. Apart from various technical problems, the type of material has been proven to be of considerable importance for the functional and histological outcome, particularly for long-term implantation. Whereas the advantageous application of low-weight, large-pore meshes based on multifilaments can be stated without doubt, it is still open whether similar results can be achieved on the basis of pure monofilaments. In the present study, a low-weight polypropylene mesh (LW) made purely of monofilaments was compared to a common heavy-weight polypropylene mesh (HW) in regard to the functional consequences and the tissue response. After implantation in rats as an inlay for 3--90 days, the abdominal-wall mobility of the implant region was recorded by 3D stereography, and the tensile strength of both the suture zone and the mesh was measured. The morphometry of the histological reaction in regard to the inflammatory infiltrate, the connective tissue, the number of granulocytes, macrophages, and fibroblasts served to reflect the tissue response. As parameters for the remodeling process at the interface the cellular activation was evaluated by TUNEL (DNA-strand breaks or apoptosis, respectively), Ki 67 (cell proliferation), and HSP 70 (cell stress). The measured tensile strength of the LW mesh confirmed a sufficient strength of the material-reduced mesh modification. After implantation the consecutive impairment of the abdominal wall mobility was reduced compared to the HW mesh, concomitant to the reduced fibrotic level at the interface. At the end of the observation period the foreign body reaction was significantly lowered for the LW mesh, almost reaching physiological values. In conclusion, these data confirm the improved biocompatibility of material-reduced mesh implants. PMID- 11870646 TI - Characterization of damage modes in dental ceramic bilayer structures. AB - Results of contact tests using spherical indenters on flat ceramic coating layers bonded to compliant substrates are reported for selected dental ceramics. Critical loads to produce various damage modes, cone cracking, and quasiplasticity at the top surfaces and radial cracking at the lower (inner) surfaces are measured as a function of ceramic-layer thickness. It is proposed that these damage modes, especially radial cracking, are directly relevant to the failure of all-ceramic dental crowns. The critical load data are analyzed with the use of explicit fracture-mechanics relations, expressible in terms of routinely measurable material parameters (elastic modulus, strength, toughness, hardness) and essential geometrical variables (layer thickness, contact radius). The utility of such analyses in the design of ceramic/substrate bilayer systems for optimal resistance to lifetime-threatening damage is discussed. PMID- 11870647 TI - Effect of additives on the microstructure and thermal properties of a mica-based glass-ceramic. AB - Previous work has shown that lithium mica glass-ceramics were excellent potential candidates as dental ceramics. The purpose of this study was to evaluate the effect of various additives on the microstructure and thermal properties of a mica glass-ceramic in the system Li(2)O-K(2)O-SiO(2)-MgO-F. Five glass compositions were prepared: a base composition and four compositions with various additives. The compositions were melted at 1400 C for 2 h and cast into 50 x 8 mm ingots. Differential thermal analyses were performed on the glasses up to 1400 C. Bars (4 x 8 x 25 mm) were cut from the ingots and heat treated at various temperatures. The crystalline phases were analyzed by x-ray diffraction. The microstructure was investigated by scanning electron microscopy. The percent crystallinity, crystal density, and average particle size were calculated from stereology measurements. X-ray diffraction revealed that the major crystalline phase was taeniolite for all glass-ceramics. Differential thermal analyses showed that the crystallization exotherm occurred in the temperature range 600--700 C. The addition of calcium fluoride was most efficient in promoting the growth of mica crystals. Larger mica plate diameter could be beneficial to the crack propagation resistance of the glass-ceramic. The addition of aluminum phosphate promoted nucleation rather than crystal growth and led to a significantly lower glass transition temperature and crystallization exotherm temperature. PMID- 11870648 TI - The effect of processing conditions on the properties of poly(methyl methacrylate) fibers. AB - Brittle failure of bone cement remains a problem for the long-term stability of hip prostheses. Fibers have been developed from poly(methyl methacrylate) (PMMA) that retain the chemistry of bone cement, but improve the mechanical properties greatly. To fabricate the fibers, a polymer melt is extruded out of a small die (spinneret) and pulled onto a take-up wheel. Varying the speed of extrusion and temperature of the melt can control the viscosity of the molten polymer. This study examines the effect of melt viscosity on the resultant properties of fibers fabricated from PMMA. The goals are to optimize fiber processing and determine processing-structure--property relationships. Resultant fibers had moderate to high levels of retained molecular orientation, and ultimate tensile strengths (UTS) ranging from 60 to 225 MPa, moduli from 1.5 to 3.5 GPa, and strain to failure from 10 to 40%. Fibers fabricated at a constant viscosity and draw velocity had identical properties, whereas decreasing the viscosity generally increased the mechanical properties and retained orientation. Linear regression models were constructed to predict how the processing variables affect the structure (orientation) of the fiber and how the structure affects the UTS. This can be used to design efficient processing methods for PMMA fibers. PMID- 11870649 TI - Macroporous poly(L-lactide) scaffold 1. Preparation of a macroporous scaffold by liquid--liquid phase separation of a PLLA--dioxane--water system. AB - A biodegradable poly(L-lactic acid) (PLLA) macroporous scaffold with a regular and highly interconnected structure in the size range from 50 to 150 mu m was fabricated from a PLLA--dioxane--water ternary system with the use of the thermally induced phase separation (TIPS) process. The phase diagram of PLLA with molecular weight above 200,000 was measured. It was found that a small change in the water content in the solvent caused a large shift in the cloud-point temperature. The porous morphology of the scaffold was closely related to the quenching route and formulation parameters, including polymer concentration, quenching temperature, aging time, and solvent composition of the ternary system. The porous morphology development in the scaffold was recorded as a function of aging time by scanning electronic microscopy (SEM). For systems with lower polymer concentrations (<4.5 wt%), polymer sedimentation occurred in the later stages of phase separation. A slight increase in the water content of the solvent mixture caused the sedimentation boundary to expand to higher polymer concentration. For systems with higher polymer concentrations (> or = 4.5 wt%), the development of phase separation was restricted by gelation that resulted from the crystallization of the PLLA chains. This gelation effect was greater at high polymer concentrations and low quenching temperatures. The macroporous expected scaffold could be optimized from the slow development of phase separation during the long coarsening process. PMID- 11870651 TI - Mechanical properties of newly developed loop ligament for connection between the EndoButton and hamstring tendons: comparison with Ethibond sutures and Endobutton tape. AB - A new artificial loop ligament (LL) without mechanical weak points, such as a knot, has been developed to connect the EndoButton and hamstring tendons for anterior or posterior cruciate ligament (CL) reconstruction. This study assessed the mechanical properties of the LL and sutures (No. 5 Ethibond thread, EndoButton tape) and those of the different CL constructs with these materials and fixation devices commonly used in CL reconstruction. The LL had a significantly higher ultimate tensile load (UTL) and stiffness value (p < 0.001) than No. 5 Ethibond thread or EndoButton tape in static and fatigue tests (50- 500 N, 1.08 million cycles). The LL-tendons-Leeds-Keio artificial ligament-staple construct complex also had significantly higher UTL and stiffness value than the Ethibond loops or EndoButton tape loop-tendons-Ethibond sutures-postscrew-fixed construct complex. Furthermore, the LL construct complex elongated the least after cyclic loading (50--150 N, 90,000 cycles). These findings suggested that using the LL construct complex might avoid a bungee cord effect and elongation before biological healing between the construct and bony socket. The LL construct complex could be a safe substitute for use during the postoperative period. PMID- 11870650 TI - A new electrochemically graded hydroxyapatite coating for osteosynthetic implants promotes implant osteointegration in a rat model. AB - Hydroxyapatite (HAP) is widely used as an osteoconductive coating for orthopedic implants. So far standard coating methods like plasma spraying produce a relatively thick coating layer (>30 microm). In addition, the chemical structure of the HAP may be altered because of the heating throughout the coating process. This may have negative effects on the coating stability, implant fixation, and induction of bone formation. The relatively thick layer may detach from the implant with the risk of wear debris. In the present study the potential of a newly developed HAP coating of implants on osteointegration was investigated in a rat model. The coating method, based on an electrochemical process, is applied in a graded manner and results in a biodegradable HAP coating with a thickness of approximately 2 mum. Coated versus uncoated titanium Kirschner wires (1.4-mm diameter) were inserted into the medullary cavity of the right femora of 5-month old female Sprague Dawley rats (n=36) in a retrograde fashion. Throughout an experimental period of 2 months the osteointegration was traced radiologically. After this time the animals were sacrificed and the implant integration was tested biomechanically with the use of a push-out test. To analyze the bone implant interface, histological sections (80 mum) were investigated with an image analyzing system. The biomechanical testing revealed a significantly higher implant fixation in the group treated with the HAP-coated implant (shear strength: 27.8 +/- 6.7 MPa) compared to control (shear strength: 8.08 +/- 3.4 MPa). The histological analyses demonstrated a better ingrowth of the implants in the HAP group with significantly more direct bone-implant contacts compared to the control group. The results demonstrate that the HAP coating promotes implant osteointegration in a rat model. PMID- 11870653 TI - Rheological properties of acrylic bone cement during curing and the role of the size of the powder particles. AB - A dynamic compressive rheometric technique was used to determine the true or complex viscosity (eta*) of three poly (methyl methacrylate), PMMA-based bone cement formulations (one commercially available and two experimental), as a function of the time that elapsed from commencement of hand mixing of the cement constituents (t). For each cement, two rheological parameters [namely, the time of onset of cure (t(ons)) and the critical cure rate (CCR), which is herein defined as the complex viscosity rate computed at t(ons)] were determined from the eta*-t data. For each cement, particle analysis was used to obtain the powder particle size distribution, from which the following parameters were obtained: (a) the overall mean particle diameter D(m), and (b) the relative amounts of small-sized PMMA beads (mean diameter d between 0 and 40 mum) (alpha) and large sized PMMA beads (d > 75 mum) (beta). It is seen that the key particle parameter is not D(m) per se but alpha and beta. Thus, the highest values of t(ons) and CCR were obtained from a cement with the highest values of alpha and beta. An explanation for these trends is given, and two areas for further research work in this field are outlined. PMID- 11870652 TI - Healing process induced by three composite prostheses in the repair of abdominal wall defects. AB - The present study compared the performance of three composite prostheses used to repair abdominal wall defects in rabbits. Two of them [Parietex Compositereg (PC) and Composixreg (CS)] are commonly used in clinical practice and one was designed by the present team (PL-PU99). At 14 and 90 days postimplant, specimens were obtained for morphological, macrophage response (RAM-11) and morphometric and biomechanical analysis. The prosthetic area covered by adhesions was significantly greater (p < 0.05) in the CS group (6.83 plus minus 2.31 cm(2)) than in PC (0.11 +/- 0.02 cm(2)) or PL-PU99 (0.10 +/- 0.07 cm(2)). At 14 days, it was observed a homogeneous, organized, well-vascularized neoperitoneum that was significantly thicker (p < 0.05) in PL-PU99. Except in the CS implants, this layer was covered by a continuous mesothelium. All three composites achieved good recipient tissue integration. Highest macrophage levels were recorded at 14 days with significantly higher values in the PL-PU99 prosthesis. Biomechanical strength was significantly greater (p < 0.05) in CS at two weeks postimplant, but it was similar at 90 days. These findings suggest that the three composites show ideal integration with host tissue, along with similar biomechanical strength at 90 days, and significantly higher adhesion formation is induced by the CS prosthesis, possibly due to incomplete mesothelialization of the lower prosthetic surface. PMID- 11870655 TI - Thermal analysis of dental resins cured with blue light-emitting diodes (LEDs). AB - Thermal analysis was used to measure the characteristics of dental resins cured with the use of a new light-activation unit equipped with high illuminant blue light-emitting diodes (LEDs). The characteristics were compared with those of resins cured with the use of two conventional halogen lamp units. The prepared base monomer consisted of a mixture of Bis-GMA and TEGDMA (60:40 by weight), with 0.5 wt% CQ/DMPT or CQ/DMAEMA. The two experimental visible-light-cured resins were polymerized for 40 s. Differential scanning calorimetry (DSC) was used to examine the thermal characteristics of the cured resins. The activation energy for the decomposition of the dental resin was calculated from the peaks of the endothermic curves obtained when the specimens were heated at three different rates (5, 10, and 15 C/min) during DSC. The activation energies calculated for the LED-cured specimens were more than 220 kJ/mol; specimens cured with the use of the halogen units had activation energies of less than 192 kJ/mol. The Knoop hardness number (KHN) of the same specimens was measured, and was higher with the blue LED units than with halogen lamp units. Therefore, dental resins cured using blue LEDs have a higher degree of polymerization and more stable three dimensional structures than those cured with halogen lamps. PMID- 11870654 TI - Bioactivity and osseointegration study of calcium phosphate ceramic of different chemical composition. AB - Synthetic hydroxyapatite (HA) and tricalcium phosphate (TCP) are promising bone substitute materials in the orthopaedic and dental fields, as their chemical composition is similar to that of bone. This study investigated the osseointegration performance of carbonated biphasic calcium phosphate (CBCP) ceramics containing carbonated hydroxyapatite and tricalcium phosphate prepared by microwave irradiation, in femoral defects of dogs. The defects were created as 3-mm holes on the lateral aspect of femur and filled with the implant material. The serum was collected postoperatively and biochemical assays for alkaline phosphatase activity levels were carried out. The animals' defective sites were radiographed at 4, 8, and 12 weeks. The radiographic results showed that the process of ossification started after 4 weeks and the defect was completely filled with new bone after 8 weeks. Histological examination of the tissue showed the osteoblastic activity inducing the osteogenesis in the defect. The complete haversian system with osteoblastic and osteoclastic activity and bone remodelling process were observed after 12 weeks. The alkaline phosphatase activity levels also correlated with the formation of osteoblast cells. This calcium phosphate ceramic has proved to work well as a biocompatible implant and as an osteoconductive and osteoinductive material for the filling of bone defects. PMID- 11870656 TI - Cytotoxicity of precious and nonprecious alloys--experimental comparison of in vitro data from two laboratories. AB - The aim of this investigation was to evaluate and compare the reproducibility of cytotoxicity data generated in two different laboratories using the same testing protocols. A series of dental alloys that are widely used in both countries were chosen. These alloys (five precious, two nonprecious) were wet ground up to 1200 grit SiC, sterilized in 70% ethanol, and extracted in sterile culture medium for 7 days. Pure copper was used as a positive control and Teflonreg and media only were used as negative controls. Test and control samples were randomized and blinded to each laboratory. Cells, primary human gingival fibroblasts, and immortalized 3T3 fibroblasts, were exposed to the extracts for 24 h. Extract cytotoxicity was evaluated spectrophotometrically with the use of a mitochondrial enzyme activity assay. Data were collected from both laboratories, combined, and subjected to a mixed-model analysis of variance. No statistical difference was obtained for the immortalized 3T3 cells, except for two extracts in which differences between the two labs were significant but were still not cytotoxic. Furthermore, no statistical differences were found for the primary cells. These data strongly suggest that cytotoxicity tests performed in different laboratories with the use of the same test materials may lead to comparable results if sample preparation, cells, test procedures, and data analyses are carefully considered. PMID- 11870657 TI - Permeation of protein from porous poly(epsilon-caprolactone) films. AB - The objective of this study was designed to extend the application of poly(epsilon-caprolactone) (PCL) in delivery of macromolecular proteins. The strategy applied here is to create a porous structure in PCL films in order to control the diffusion rate of protein. Various amounts of both high-molecular weight and low-molecular-weight poly(ethylene glycol) (PEG) were used as pore forming agents. The porous films were prepared by a solvent-casting-leaching method. The thicknesses of the prepared films were controlled to be in the range of 75.3 +/- 0.6 similar 81.7 +/- 0.6 mum. The pore fraction of films was determined to be 27.7 +/- 1.0% similar 52.5 +/- 0.8% for PEG(10000) and 26.6 +/- 1.8% similar 48.8 +/- 1.4% for PEG(4000). The pore fraction initially increased with increasing amounts of PEG, independent of the molecular weight of PEG. In the permeation study, lysozyme was used as a model diffuser. The permeation rate of protein increased as the pore fraction of films increased, especially when 30 similar 40% of PEG was added initially, and this phenomenon was more prominent when low-molecular-weight PEG was used. This result was probably due to the highly porous structure creating interconnected channels in the films, further enhancing protein diffusion. In addition, the size of micropores formed by PEG(4000) was observed to be larger than by PEG(10000), which also accounted for faster permeation rate of lysozyme through PCL-PEG(4000) porous films. PMID- 11870658 TI - Flexural strength distribution of a PMMA-based bone cement. AB - Polymethylmethacrylate bone cement containing either no added antibiotic or 0.5 g of Gentamicin was prepared and stored either in air at room temperature or in a 37 degree C water bath for 48 h. An additive-free cement stored in air at room temperature was also tested for purposes of comparison. Following storage the specimens were tested in flexure. Weibull statistics demonstrated to fit the flexural strength distribution of all the materials tested with regression coefficients of at least 0.98. The presence of a BaSO(4) radiopacifier markedly reduced the mean flexural strength and increased the data scatter in the air stored specimens. On the other hand, the flexural strength of both impregnated and nonimpregnated antibiotic increased when those materials were stored in water at 37 degree C, compared with the same material stored in air, as a consequence of the water ingress. The water-stored antibiotic-impregnated cement displayed lower flexural strength, increased data scatter, and a remarkably higher number of weak specimens compared with the antibiotic-free cement. The influence of the load type on the flexural behavior was studied by testing the air-stored specimens in three-point bending and four-point bending. Cements tested in four point bending resulted in lower flexural strength than that tested in three-point bending. The ratio of mean strength measured in the different load arrangements was satisfactory, as predicted by the Weibull model. PMID- 11870659 TI - Bioactive glass as bone-graft substitute for posterior spinal fusion in rabbit. AB - Bioactive glass S53P4 and autogenous bone were studied as bone graft materials for spinal fusion in a rabbit model. Sixteen rabbits underwent surgery by a dorsal approach. A bioactive glass, a combination of bioactive glass and autogenous bone (70/30 vol%), and autogenous bone were implanted at two thoracolumbar vertebraes for 4 and 12 weeks. The volume, consolidation to vertebrae, and fusion of the graft material were evaluated with plain-film radiology, computed tomography (CT) and bone-mineral density measurements, and compared with histomorphometrical measurements. Radiological consolidation by CT of bone graft to underlying vertebrae at 12 weeks was observable in all groups. This was histologically confirmed as bone was growing from the vertebrae into the graft material. Radiologic fusion of vertebraes was, at 12 weeks, observable in all groups in 50--75% of the cases. The radiologic fusion seen at the CT scans could, however, not be confirmed by histology in any of the three groups. Significant differences for graft material and observation period with the use of bone-mineral density measurements (Hounsfield units) were also observable, with the highest measured values for the bioactive glass group and the lowest for the autogenous bone group. The results indicate that bioactive glass have potential as bone-graft material in spinal fusion. The reliability of radiologic evaluation methods in spinal surgery using bone substitutes is also questioned and discussed. PMID- 11870660 TI - Prophylactic surgery in hereditary cancer syndromes: an ounce of prevention may be the only cure. PMID- 11870661 TI - Management of well-differentiated thyroid carcinoma presenting within a thyroglossal duct cyst. AB - BACKGROUND AND OBJECTIVE: Well-differentiated thyroid carcinoma (WDTC) is diagnosed in approximately 1.5% of thyroglossal duct cysts (TGDC). No clear consensus exists regarding further management after adequate excision of the cyst, especially the role of total thyroidectomy and postoperative radioactive iodine therapy. The current review was undertaken in an attempt to clarify these issues. METHODS: Demographic, clinical, tumor, treatment, pathology, and outcome data on 57 eligible patients reported in recent literature were pooled together with 5 patients treated at our institution for this analysis. RESULTS: A Sistrunk operation was performed for resection of the thyroglossal duct cyst in the majority (90%) of patients. Histologic examination of the tumor in the cyst revealed that papillary carcinoma was the most frequent (92%) histologic type. A total thyroidectomy was performed consequent to the diagnosis of thyroglossal duct cyst carcinoma in approximately half of the 62 patients. A malignant tumor was reported in 27% of the thyroidectomy specimens. Postoperative radioactive iodine therapy was administered in 16 (26%) patients. With a median follow-up of 71 months (range 1-456 months), the 5- and 10-year Kaplan-Meier overall survival was 100 and 95.6%, respectively. There were no disease-related deaths reported in any of the patients. Univariate analysis revealed that the only significant predictor of overall survival was the extent of primary surgery for the thyroglossal cyst. The addition of total thyroidectomy to Sistrunk operation did not have a significant impact on outcome (P = 0.1). Patients treated with postoperative radioactive iodine (RAI) fared significantly worse than those that did not need RAI, which may be explained by the fact that this modality would generally be used in patients with higher risk tumors. CONCLUSIONS: The Sistrunk operation is adequate for most patients with incidentally diagnosed TGDC carcinoma in the presence of a clinically and radiologically normal thyroid gland. Results of adequate excision using the Sistrunk operation are excellent and the concept of risk-groups should be used to identify patients, who would benefit from more aggressive treatment. PMID- 11870663 TI - Cellular manifestations of human papillomavirus infection in laryngeal tissues. AB - BACKGROUND AND OBJECTIVES: Although epidemiologic studies have suggested human papillomavirus (HPV) to be an etiological agent in laryngeal carcinogenesis, little is known on the cellular manifestations of HPV infection in these tumors. In this study, we investigated the frequency of HPV infection in various neoplastic and non-neoplastic laryngeal tissue and its association with expression of the proliferating cell nuclear antigen (PCNA) and the tumor suppressor protein p53. METHODS: Tissues were analyzed by polymerase chain reaction (PCR) for the presence of HPV and by immunocytochemistry for the expression of p53 and PCNA. RESULTS: None of the six normal laryngeal tissues showed the presence of HPV. Thirteen out of the 16 papillomas were positive for HPV, while 15 out of the 44 invasive cancers were HPV positive. PCNA expression increased as the lesion progressed through increasing histological abnormality (r = 0.64400, P = 0.00000). The correlation between the type of laryngeal neoplasm and p53 accumulation was significant (r = 0.54839, P = 0.00000). Significant correlation was also evident between presence of HPV and p53 accumulation (r = 0.34259, P = 0.00424) and PCNA expression (r = 0.036024, P = 0.00266) indicating that HPV positive tumors showed significant p53 accumulation and increased proliferation. There was also correlation between p53 and PCNA expression (r = 0.67475, P = 0.00000) indicating that in all tumors with p53 accumulation, there was a corresponding increase in PCNA expression. CONCLUSIONS: The results suggests that changes in p53 and PCNA expression may be associated with HPV infection, and could play a role in laryngeal carcinogenesis. PMID- 11870664 TI - Early and late morbidity associated with axillary levels I-III dissection in breast cancer. AB - BACKGROUND AND OBJECTIVES: Axillary dissection may cause substantial morbidity in breast cancer patients. The purpose of this study was to investigate the value of a registration method of morbidity of the arm and shoulder, which is frequently used by surgeons and which includes the measurement of range of movement, strength, and pain. METHODS: We surveyed 148 patients who had received an axillary dissection as part of breast cancer surgery. Of these patients, 77 had undergone axillary dissection 6-12 months ago and 71 patients more than 5 years ago. In all patients, an objective measurement of shoulder movement and a subjective measurement of pain and arm strength was performed. RESULTS: A difference of more than 20 degrees in abduction, ventral elevation, or dorsal elevation occurred in 12% of the patients. Pain or loss of strength were measured in half of the patients. Shoulder movement, pain, and arm strength were not significantly different between the patients who underwent mastectomy or breast conserving surgery. Also, no significant difference could be found in shoulder movement, pain, and arm strength between the patients who underwent axillary dissection 6-12 months ago and those who underwent it more than 5 years ago. CONCLUSIONS: Pain, loss of arm strength, and limitation of shoulder movement are frequent complaints after axillary dissection for breast cancer and appear to be independent of the length of follow-up and the type of surgery (i.e., breast conservation or mastectomy). PMID- 11870666 TI - Use of microsatellite analysis in young patients with colorectal cancer to identify those with hereditary nonpolyposis colorectal cancer. AB - BACKGROUND AND OBJECTIVES: The frequency of microsatellite instability (MSI) in young patients with colorectal cancer was evaluated, including reexamination of the medical and family history of each patient, and interviews with the patients to determine any possible new occurrence of hereditary nonpolyposis colorectal cancer (HNPCC) in the patients themselves or their family members. METHODS: Fifty three young patients (younger than 40 years of age) with colorectal cancer were selected and investigated. DNA was extracted from paraffin sections and microsatellite analysis was performed. RESULTS: The frequency of MSI among the young patients with colorectal cancer was 50.9%, which was significantly higher than the rate of 12-21% noted in older patients with colorectal cancer (P < 0.001). For the 24 young patients with colorectal cancer who did not have MSI, only one case of HNPCC kindred and two cases with a family history of cancer were identified. In contrast, among the 20 young patients with colorectal cancer who had MSI, five cases of HNPCC kindred, two cases with metachronous patients with colorectal cancer, and three cases with a family history of cancer were identified. CONCLUSION: Our results suggest that a defect in the DNA mismatch repair system may play some role in carcinogenesis in young patients with colorectal cancer. Microsatellite analysis and subsequent interviews regarding medical and family history are useful tools for efficiently identifying possible cases of HNPCC among young patients with colorectal cancer. PMID- 11870667 TI - A multi-institutional study of immunohistochemical investigation for the roles of cyclin D1 and E-cadherin in superficial squamous cell carcinoma of the esophagus. AB - BACKGROUND AND OBJECTIVES: Aiming to clarify and possibly extend indications for minimally invasive treatment, we characterized superficial esophageal cancers (SEC) with respect to biologic properties regulating malignant potential. METHODS: Surgical specimens obtained at eight cancer institutes from 222 Japanese patients with SEC (all squamous cell carcinomas) were investigated immunohistochemically for expression of cyclin D1 and E-cadherin. RESULTS: Perturbations of cyclin D1 (overexpression) and E-cadherin (reduced expression) were observed in 37.6% (68 of 181) and 39.9% (71 of 178) of SEC patients. E cadherin expression was more frequently reduced in cancers that invaded the submucosal layer, while cyclin D1 overexpression was constant, irrespective of depth of invasion. Overexpression of cyclin D1 was more frequent in poorly differentiated squamous cell carcinomas, while E-cadherin did not vary according to histologic differentiation. Lymph node metastasis, the only independent postoperative prognostic factor in these patients, occurred in only 4.8% of mucosal cancers (2 of 42), but in 51.1% of submucosal cancers (92 of 180). However, neither cyclin D1 nor E-cadherin status affected lymph node metastasis. CONCLUSION: Both E-cadherin and cyclin D1 play important roles in esophageal carcinogenesis, but neither can be used to identify patients who do not require lymph node dissection and might be treated by endoscopic mucosal resection. PMID- 11870668 TI - Immunohistochemical study of receptor activator of nuclear factor kappa-B ligand (RANK-L) in human osteolytic bone tumors. AB - BACKGROUND AND OBJECTIVES: Osteolytic bone tumors produce intercellular signaling proteins that regulate bone remodeling by altering the rates of osteoclast and osteoblast differentiation and activity. This report examines osteolytic bone tumor expression of receptor activator of nuclear factor B-ligand (RANK-L), a cytokine that is arguably the most critical regulator of osteoclast differentiation and activation. METHODS: This prospective immunohistochemical study examined RANK-L expression in frozen tissues from sixteen surgical specimens of patients who underwent surgery for the treatment of osteolytic bone tumors between 1999 and 2000. RESULTS: RANK-L was positive in 13 of the 16 cases. Primary benign bone tumors, primary malignant bone tumors, and metastasis to bone were positive for RANK-L. CONCLUSIONS: The cells in some, but not all, osteolytic tumors produce the cytokine RANK-L. Further study is necessary to determine in which specific tumors RANK-L is the cytokine responsible for increased osteoclastic activity, and to develop possible therapeutic use of RANK-L antagonists such as osteoprotegerin (OPG). PMID- 11870669 TI - Regression of metastatic carcinoid tumors with octreotide therapy: two case reports and a review of the literature. AB - BACKGROUND: The antiproliferative effect of the somatostatin analogue, octreotide, on metastatic carcinoid tumors is poorly understood. Partial tumor regression seen radiographically has been reported with the use of octreotide therapy for neuroendocrine tumors. Complete regression of carcinoid tumors is rarely reported. RESULTS: Two patients with metastatic midgut carcinoid tumors were treated with subcutaneous octreotide 300 microg/day for symptomatic control of their carcinoid syndrome before debulking palliative surgery. During the laporatomies, both patients were found to have complete macroscopic regression of the metastatic lesions that had been identified radiologically before surgery, including liver metastases in one patient and periportal and retrocaval lymph nodes in the other. After surgery, the patients were evaluated every 3 months, and had no detectable disease at 30 and 43 months, respectively. Thirty cases of partial tumor regression with octreotide administered with or without other treatment modalities have been reported in the literature. Most of the patients involved received other treatment modalities. Only one other case reported in the literature showed complete regression with octreotide monotherapy. CONCLUSIONS: We report two cases of metastatic midgut carcinoid tumors that demonstrated a significant anti-proliferative response to octreotide monotherapy. Review of the literature failed to identify any specific prognostic factors with which the response to octreotide can be predicted. Possible mechanisms for this antiproliferative effect of octreotide on carcinoid tumors are discussed. PMID- 11870670 TI - Gastrointestinal cancer and sentinel node navigation surgery. AB - In the twentieth century, lymph node dissection based on metastatic distribution has developed as the standard procedure for surgical management of gastrointestinal cancer [1]. The fear that invisible micrometastases might be present has encouraged aggressive resection with lymphadenectomy to control the disease. However, the prognostic benefits of extensive surgery are still unknown and the universal application of radical surgery may affect surgical morbidity, mortality, and quality of life after surgery, particularly in patients without lymphatic spread. In the twenty-first century, a novel technology to detect micrometastases without extensive surgical resection is required to establish an individualized surgical management approach to gastrointestinal cancer. Lymphatic mapping techniques are now used in the control of superficial malignancies, such as malignant melanoma and breast cancer, providing a novel tool that may also be of use for gastrointestinal cancers. PMID- 11870671 TI - En bloc pancreaticoduodenectomy for right colon cancer invading adjacent organs. AB - BACKGROUND AND OBJECTIVE: Surgical treatment of colorectal cancer needs an extended resection of the tumor en block with invaded organs. There is little information about the surgical treatment of right-sided colon carcinoma directly involving duodenum and pancreas. Our objective is to report our experience with three patients who underwent en bloc pancreaticoduodenectomy and hemicolectomy for locally advanced right colon cancer. METHODS: Retrospective review of clinical records of patients with colon cancer. RESULTS: Three patients with right colon cancer adherent to adjacent organs underwent en block surgery. No operative deaths occurred. All patients are alive without evidence of disease at 10, 30, 113 months of follow-up, respectively. CONCLUSION: Locally advanced right sided colon cancer can be safely treated with en bloc pancreaticoduodenectomy and colectomy with excellent long-term results. PMID- 11870673 TI - Massive bleeding from duodenal carcinoid treated by gastroduodenal artery embolization. PMID- 11870674 TI - Long intramedullary stems of prosthetic components reduce complications, when allograft-prosthesis composite is used in tumor reconstruction. PMID- 11870676 TI - Flunisolide HFA vs flunisolide CFC: pharmacokinetic comparison in healthy volunteers. AB - Two preparations of flunisolide, an inhaled corticosteroid, were compared in a parallel, multiple-dose study of 31 healthy volunteers. The new flunisolide preparation substitutes hydrofluoroalkane (HFA) for chlorofluorocarbon (CFC) as a propellant and incorporates a spacer into its pressurized metered-dose inhaler (pMDI). In this study, subjects were randomly assigned to receive flunisolide CFC 1000 microg bid; flunisolide HFA 170 microg bid; or flunisolide HFA 340 microg bid. Dosing was continued for 13.5 days. Plasma samples were analyzed after the first dose on day 1 and again after 13.5 days of treatment. No significant differences in day 1 dose-adjusted peak plasma concentrations (C(max)) were observed. Dose proportionality in C(max) and area under the concentration--time curves (AUC) was observed for the flunisolide HFA 170 and 340 microg bid groups on days 1 and 14. Day 1 mean dose-adjusted AUC was significantly greater in the flunisolide CFC 1000 microg bid group than in either flunisolide HFA group, indicating greater systemic availability of flunisolide CFC. Oral clearance and volume of distribution were significantly higher for flunisolide CFC than for flunisolide HFA. This may be due to greater oropharyngeal deposition by the flunisolide CFC formulation. Another indicator of greater flunisolide CFC oropharyngeal deposition was observed in C(max) and AUC(0--tlast) values for 6beta-OH flunisolide, the first-pass metabolite of flunisolide. The values of these pharmacokinetic parameters were significantly higher in the flunisolide CFC group than in the 340 microg bid flunisolide HFA group on days 1 and 14. However, this was not the case for cortisol values where flunisolide HFA accounted for less oropharyngeal deposition and more targeted delivery without adverse events. The study demonstrated that flunisolide HFA administered through a pMDI with built-in spacer was safe and well tolerated in healthy volunteers. PMID- 11870677 TI - Amlodipine bioequivalence study: quantification by liquid chromatography coupled to tandem mass spectrometry. AB - OBJECTIVE: To assess the bioequivalence of two amlodipine tablet formulations (Amlodipine 5 mg tablet from Merck S.A. Industrias Quimicas, Brazil as test formulation and Norvasc 5 mg tablet from Laboratorios Pfizer Ltd., Brazil as reference formulation) in 24 healthy volunteers of both sexes. METHODS: The study was conducted using an open, randomized two-period crossover design with a 4-week washout interval. Plasma samples were obtained over a 144 h period. Plasma amlodipine concentrations were analyzed by combined liquid chromatography coupled to tandem mass spectrometry (LC-MS-MS) with positive ion electrospray ionization using multiple reaction monitoring (MRM). From the amlodipine plasma concentration vs time curves, the following pharmacokinetic parameters were obtained: AUC(last), AUC(0-inf) and C(max). The statistical interval proposed was 80-125% according to the US Food and Drug Administration Agency. RESULTS: The limit of quantification was 0.1 ng/ml for plasma amlodipine analysis. The geometric mean and the 90% confidence interval (CI) test/reference ratios were 101.2 (92.9-110.2%) for AUC(last), 99.6 (91.5-108.4%) for AUC(0-inf) and 98.5 (89.0-109.1%) for C(max). CONCLUSION: Since the 90% CI for AUC(last), AUC(0-inf) and C(max) ratios were within in the 80-125% interval proposed by the US FDA, it was concluded that Amlodipine 5 mg tablet (test formulation) was bioequivalent to Norvasc 5 mg tablet, in terms of both rate and extent of absorption. PMID- 11870678 TI - A critical evaluation of the brain efflux index method as applied to the nitric oxide synthase inhibitor, aminoguanidine. AB - The Brain Efflux Index (BEI) method is an in vivo procedure designed to quantitate saturable efflux mechanisms resident at the blood--brain barrier (BBB). The present work utilized the BEI method to assess the BBB efflux mechanisms of [(14)C]aminoguanidine, a nitric oxide synthase inhibitor. The BEI for [(14)C]aminoguanidine was >100% (relative to [(3)H]inulin diffusion) over a range of 41-184 pmol after 40 min. The unusually high retention (>100%) of [(14)C]aminoguanidine suggested brain parenchymal sequestration, either by neuronal uptake or tissue protein binding. The uptake of [(14)C]aminoguanidine in dendritic neuronal endings (synaptosomes) showed a saturable concentration dependency, consistent with a carrier-mediated process. Nonlinear least-squares regression yielded the following Michaelis--Menten and diffusional (k(ns)) parameters for synaptosomal [(14)C]aminoguanidine uptake: V(max)=118.50 +/- 28.77 pmol x mg protein(-1)/3 min; K(m)=58.34 +/- 8.33 muM; k(ns)=0.15 +/- 0.029 pmol x mg protein(-1)/3 min/muM; mean +/- SEM; n=3 concentration profiles). Protein binding studies using brain tissue showed negligible binding. In summary, this work identified three principle findings: (1) An apparent lack of quantifiable aminoguanidine BBB efflux; (2) a previously undescribed synaptosomal accumulation process for aminoguanidine; and (3) an interesting limitation of the BEI technique where unusual brain parenchymal sequestration yields values >100%. PMID- 11870679 TI - Effect of smoking on single dose pharmacokinetics of phenobarbital. AB - In order to determine interaction of smoking with single dose pharmacokinetics of phenobarbital, a 60 mg tablet of the drug was given to 12 healthy male subjects in two groups (6 smokers and 6 non-smokers) in a double blind study. An HPLC method using UV detection was developed to assess phenobarbital in plasma of the subjects. Pharmacokinetic parameters were calculated and compared in the two groups. Pharmacokinetic parameters of the two groups were not significantly different in the two groups (p<0.05). The results show no considerable effect of cigarette smoking on phenobarbital pharmacokinetics, which is in agreement with enzyme studies performed previously. PMID- 11870680 TI - Reporting clinical trials: full access to all the data. PMID- 11870681 TI - Familial amyotrophic lateral sclerosis. AB - The increasing complexity of the pathways implicated in the pathogenesis of familial amyotrophic lateral sclerosis (ALS) has stimulated intensive research in many directions. Genetic analysis of familial ALS has yielded six loci and one disease gene (SOD1), initially suggesting a role for free radicals in the disease process, although the mechanisms through which the mutant exerts toxicity and results in selective motor neuron death remain uncertain. Numerous studies have focused on structural elements of the affected cell, emphasizing the role of neurofilaments and peripherin and their functional disruption in disease. Other topics examined include cellular homeostasis of copper and calcium, particularly in the context of oxidative stress and the processes of protein aggregation, glutamate excitotoxicity, and apoptosis. It has become evident that there is considerable interplay between these mechanisms and, as the role of each is established, a common picture may emerge, enabling the development of more targeted therapies. This study discusses the main areas of investigation and reviews the findings. PMID- 11870682 TI - Magnetic stimulation of the central and peripheral nervous systems. AB - Since 1985, when the technique of transcranial magnetic stimulation (TMS) was first developed, a wide range of applications in healthy and diseased subjects has been described. Comprehension of the physiological basis of motor control and cortical function has been improved. Modifications of the basic technique of measuring central motor conduction time (CMCT) have included measurement of the cortical silent period, paired stimulation in a conditioning test paradigm, repetitive transcranial magnetic stimulation (rTMS), and peristimulus time histograms (PSTH). These methods allow dissection of central motor excitatory versus inhibitory interplay on the cortical motor neuron and its presynaptic connections at the spinal cord, and have proven to be powerful investigational techniques. TMS can be used to assess upper and lower motor neuron dysfunction, monitor the effects of many pharmacological agents, predict stroke outcome, document the plasticity of the motor system, and assess its maturation and the effects of aging, as well as perform intraoperative monitoring. The recent use of rTMS in the treatment of depression and movement disorders is novel, and opens the way for other potential therapeutic applications. PMID- 11870683 TI - Effects of voluntary activity on the excitability of motor axons in the peroneal nerve. AB - To investigate whether there are inter-nerve differences in the extent and pattern of axonal excitability changes produced by voluntary contractions of tibialis anterior (TA) and abductor pollicis brevis (APB), threshold tracking was used to measure axonal excitability parameters [threshold, supernormality and strength-duration time constant (tauSD)] of peroneal and median motor axons in 11 healthy subjects. Maximal contractions for 1 min resulted in an increase in threshold, an increase in supernormality, a decrease in tauSD and an increase in latency, all of which indicate axonal hyperpolarization. The increase in threshold was less in peroneal axons (18 +/- 4%) than median axons (37 +/- 6%, mean +/- SEM, P < 0.001), and was accompanied by smaller absolute changes in latency, supernormality, and tauSD. Peroneal axons had less supernormality at rest but a greater change in supernormality for the change in threshold. There were major contraction-induced changes in the compound muscle action potential of TA but not that of APB. Voluntary contractions depress axonal excitability, but the changes are quantitatively different for motor axons innervating different muscles. There are three clinical implications. First, weakness and fatigue due to activity-dependent conduction block may vary for different muscles, independent of disease severity, and therapeutic strategies to overcome activity dependent conduction block may not be equally effective for different muscles. Second, in motor control studies using the H reflex to document motoneuron excitability, a constant stimulus will not produce a constant neural volley if the stimulated axons have been activated by, for example, a voluntary contraction. Third, TA is probably not optimal for testing for activity-dependent conduction block. PMID- 11870684 TI - Cosegregation of the mitochondrial DNA A1555G and G4309A mutations results in deafness and mitochondrial myopathy. AB - We report a patient with progressive external ophthalmoplegia (PEO), exercise intolerance, and deafness after aminoglycoside exposure, harboring two pathogenic mutations in her mtDNA: an A1555G in the 12S rRNA gene and a G4309A in the tRNA(Ile) gene. Muscle histochemistry showed abundant ragged-red fibers, and biochemistry revealed normal respiratory chain function. The A1555G mutation was homoplasmic in blood from the proband and from all maternal relatives. The G4309A mutation was abundant in the proband's muscle, less abundant in her blood, still less abundant in the mother's blood, and absent in blood from other maternal relatives. Family members were asymptomatic. Our data suggest that the former mutation resulted in aminoglycoside-induced deafness and the latter caused PEO plus exercise intolerance. PMID- 11870685 TI - Bent spine syndrome: computed tomographic study and isokinetic evaluation. AB - Primary acquired lumbar kyphosis of the adult, or bent spine syndrome, is probably secondary to late-onset muscle dystrophy preferentially involving the paravertebral muscles. In order to better define the extent of this myopathy, we compared spinal computerized tomography scans and muscle force, power, and fatigability, assessed by isokinetic tests, of the spinal erector and pelvic and shoulder girdle muscles in 23 patients with primary acquired lumbar kyphosis (20 women and 3 men, mean age 72.8 years) and 15 matched control subjects. Patients had decreased spinal and shoulder muscle density. Force, power, and work done were reduced in their paravertebral muscles, whereas in the scapular and gluteus medius muscles only work done after a series of 20 rapid movements showed a decrease. Primary acquired lumbar kyphosis in the adult may be considered as a late-onset girdle myopathy with subclinical involvement of the muscles of the pelvic and shoulder girdles. PMID- 11870686 TI - Effects of leukemia inhibitory factor on rat skeletal muscles are modulated by clenbuterol. AB - Leukemia inhibitory factor (LIF) is implicated in skeletal muscle regeneration, but the effect of exogenous LIF on uninjured muscles is not known. We tested the hypothesis that LIF administration would stimulate muscle hypertrophy, with an increased effect during clenbuterol-induced fiber remodeling. Rats received daily injections of either saline or LIF, and either regular or clenbuterol supplemented drinking water for 4 weeks. In the slow-twitch soleus muscles of LIF treated rats, specific force (sP(o)) and muscle fiber size were increased by approximately 13% and approximately 26%, respectively, compared to saline-treated rats. In the soleus muscles of rats receiving LIF and clenbuterol, compared to rats receiving clenbuterol alone, maximum isometric tension (P(o)) was approximately 19% greater. LIF alone did not affect the properties of fast-twitch extensor digitorum longus (EDL) muscles, but in rats receiving LIF and clenbuterol, compared to clenbuterol alone, EDL fiber size and muscle mass were increased by approximately 20% and approximately 10%, respectively. The hypertrophic effects of exogenous LIF on uninjured skeletal muscles indicate that LIF may have application in the treatment of conditions characterized by muscle wasting. PMID- 11870688 TI - Running endurance abnormality in mdx mice. AB - The mdx mouse lacks dystrophin and has histological features of Duchenne muscular dystrophy but little weakness in the first year of life. We report here an early deficit in voluntary wheel running, as assayed with a computerized wheel. All mdx mice showed an intermittent running pattern, in contrast to the continuous running seen in controls. The average continuous running time differed significantly between mdx and control mice at all ages tested (5-21 weeks). This assay is noninvasive, has the advantage of unbiased automatic data collection, and should be useful for quantifying the mdx deficit in therapeutic studies. PMID- 11870687 TI - Acquisition and expression of proximal and distal upper limb stimulus-response curves to transcranial magnetic stimulation. AB - Motor cortex stimulus-response (S/R) curves are an indication of cortical excitability and are of relevance to topographical mapping. The aims of this study were to compare two different methods of collecting data to construct a S/R curve for transcranial magnetic stimulation (TMS) in the upper limbs, to identify reliable summary statistics for the S/R curve, and to determine whether S/R curves predicted motor threshold. Motor evoked potentials (MEP) were obtained from biceps brachii (BB) and abductor pollicis brevis (APB) muscles at rest with a circular coil centered at the vertex. Motor threshold was determined using a validated protocol. MEPs were obtained with 1% increments in intensity or from the average of five trials at 5% increments. The S/R relationships were fitted to linear, S-shaped, and Boltzmann functions. A linear function determined from the average of five trials accurately summarized our data (r2 from 0.6 to 0.9, P < 0.05, n = 8, for right APB and from 0.6 to 0.9, P < 0.05, n = 8 for right BB). The X-axis intercept of the line determined using these methods fell between the upper and lower limits of motor threshold in all eight subjects. We propose that MEP values obtained at intervals of 5% averaged over five trials, fitted to a linear function provides a practical means of assessing the S/R characteristics of TMS for proximal and distal upper limb muscles. PMID- 11870689 TI - Symmetry of nerve conduction studies in different stages of diabetic polyneuropathy. AB - Nerve conduction studies (NCS) in diabetic sensorimotor polyneuropathy (DSP) are sensitive, noninvasive, and associated with small coefficients of variation, and correlate well with underlying peripheral nerve morphological change. For these reasons, the current reference standard for DSP involves multivariate instruments that emphasize NCS results. However, the interside symmetry of NCS findings in different stages of DSP are unknown, although requirement for symmetry has been suggested in clinical trials of DSP. We therefore aimed to determine the degree of symmetry of NCS findings in DSP of differing severity stages. A cohort of diabetic patients, including patients without neuropathy and those with mild to severe DSP, was studied. We also studied a series of nondiabetic, healthy subjects. A variation of stratified sampling by means of a clinical neuropathy score ensured that a broad spectrum of neuropathy was studied. A total of 478 subjects was ascertained; patient accrual was discontinued when the smallest clinical group consisted of 50 subjects. Nerve conduction studies were conducted prospectively and in a blinded fashion using surface recordings, averaging for sensory action potentials, control of limb temperature, and standardized techniques. Median and ulnar motor and sensory, peroneal and tibial motor, and sural NCS were performed. Interside symmetry, independent of neuropathy severity, was observed for all investigated nerves, except for the median sensory nerve action potential amplitude, which was lower on the right side. These results confirm that abnormal NCS findings consistent with DSP are reliably symmetrical with the exception of the amplitude of the median sensory nerve action potential. Thus, unilateral evaluation of NCS in DSP is sufficient as a reference standard in clinical trials. We also conclude that great degrees of asymmetry in NCS results are reason to question inclusion of DSP patients in clinical trials. PMID- 11870690 TI - Acceleration by MS-818 of early muscle regeneration and enhanced muscle recovery after surgical transection. AB - The synthesized pyrimidine compound MS-818 has neurotrophic effects in several kinds of neuronal cells, but its effect with respect to muscle cells remains unknown. We therefore examined the effects of MS-818 on regeneration for 12 weeks in a wounded area (damaged and gap areas) of cut muscle in adult rats. The right semitendinosus muscles of treated and control groups were severed and sutured at the belly and the left semitendinosus muscles were left intact. MS-818 was administered intraperitoneally to the treated group at a dose of 5 mg/kg once daily. Control rats received an equal volume of physiological saline. A reference group underwent no surgical procedure. MS-818 significantly increased the maximal isometric twitch tension (Tmax) compared to control and reference rats after week 4 (approximately 1.4-fold control value; 0.6-fold reference value). Northern blotting showed that MS-818 enhanced myogenin mRNA expression to about 1.5-fold above the control level at 2, 4, and 7 days after surgery. Immunohistochemical and histochemical studies showed significant enhancement in the treated group since myogenic cells expressed desmin and were positive for neonatal myosin, and the fiber diameters and numbers of premature myofibers and end plates were increased when compared with those in the control group. These results show that MS-818 accelerated the proliferation and differentiation of activated satellite cells and the fusion of myotubes to form immature myofibers. At week 12, Tmax, fiber diameter, and number of end plates in the treatment group recovered 60, 85, and more than 100%, respectively, compared to the reference group. The mechanism of MS-818 effects on the accelerated regeneration of cut muscle is discussed. PMID- 11870691 TI - Analysis of motor pathway involvement in konzo using transcranial electrical and magnetic stimulation. AB - To elucidate the involvement of motor pathways in konzo, 21 konzo subjects (mean age 22 years) underwent transcranial electrical stimulation (TES) in 1998. Fourteen konzo subjects (mean age 21 years) underwent transcranial magnetic stimulation (TMS) in 2000. Three subjects underwent both TES and TMS. Motor evoked potentials (MEPs) were recorded in the abductor pollicis brevis (APB) muscle with TES, and in the abductor digiti minimi (ADM) and tibialis anterior (TA) muscles with TMS. APB-MEPs were normal in 2 of 21 subjects and absent in 9; central conduction time (CCT) was prolonged in 10. Resting ADM-MEPs were absent in 9 of 14 subjects with clinically preserved upper limbs. Among these nine, seven subjects responded after facilitation. Most subjects (13 of 14) failed to show TA-MEPs. Of the subjects who underwent both types of stimulation, one had normal TES-MEP but abnormal ADM-MEP with TMS. These findings suggest involvement of both corticomotoneurons and motor descending pathways in konzo. PMID- 11870692 TI - Intermuscle differences in activation. AB - The objective of this study was to investigate differences within individual subjects in the ability to activate the quadriceps, plantar flexors (PF), dorsiflexors (DF), and elbow flexors (EF) during isometric contractions. Twelve male subjects performed submaximal and maximal voluntary isometric contractions, and maximal tetanic contractions were also induced by electrical stimulation. The interpolated twitch technique was used to gauge the extent of muscle inactivation or inability to produce maximum force. Measurements included torque output, absolute and relative rate of force development (RFD), and percentage of muscle inactivation. The quadriceps exceeded all other muscle groups in voluntary and tetanic torque output, voluntary absolute RFD, and absolute and relative tetanic RFD. The quadriceps also exceeded the PF and DF in voluntary relative RFD and had greater muscle inactivation (15.5%) than the EF (5.0%), PF (5.0%), and DF (1.3%). Although the higher RFD may suggest a higher percentage of type II fibers in the quadriceps, their higher threshold of recruitment leads to greater difficulty in fully activating the quadriceps. PMID- 11870694 TI - Assessment of contractile and noncontractile components in human skeletal muscle by magnetic resonance imaging. AB - A magnetic resonance imaging (MRI) technique for the assessment of contractile and noncontractile components of human skeletal muscle is described, and the inter-rater and intra-rater test-retest reliability for repeated measurements from the same MR image are examined. Twenty cross-sectional MR images from the right lower leg were obtained from 30 healthy young men and women (mean age 24.1 years, SD 3.3). The anatomical cross-sectional area (aCSA; cm2), the cross sectional area of noncontractile components (Noncon; cm2), the contractile cross sectional area (cCSA = aCSA minus Noncon; cm2), and the relative amount of Noncon (%), of the ankle dorsiflexor muscle compartment were determined for each slice using a computer-based image analysis system. Reliability for repeated measurements of the slice with the largest aCSA for the 30 subjects was analyzed by two raters on two different occasions. Inter-rater reliability on both occasions, assessed by the intraclass correlation coefficient (ICC), was excellent for cCSA (ICC3.1 = 0.99) and Noncon (ICC(3.1) > 0.82). Intra-rater (between occasions) reliability was excellent for the two raters for measurements of cCSA (ICC1.1 = 0.99) and Noncon (ICC1.1 > 0.94). Bland and Altman analyses did not identify any clinically relevant bias in the measurements. Method errors were acceptable: within subjects coefficients of variation (CV) was less than 1.8% for cCSA and less than 16.3% for Noncon. It is concluded that repeated measurements of contractile and noncontractile components of the ankle dorsiflexor muscle compartment, obtained from the same MR image, are highly reliable. PMID- 11870693 TI - Rapidly progressive amyloid polyneuropathy associated with a novel variant transthyretin serine 25. AB - We report a 52-year-old woman with a novel transthyretin (TTR) variant serine replacing alanine at residue 25 [Ala25Ser (Serine 25)], who showed a unique clinical picture with a relatively acute onset neuropathy within a few days of an influenza vaccination, progressing to a severe degree within 2 years. Sural nerve biopsy revealed amyloid deposition in the endoneurium. Sequencing of the proband's DNA revealed a G to T transversion at the first position of codon 25 of TTR gene. DNA analysis of this family showed the same mutation in her older sister and a niece, but her parents did not have the mutation. Haplotype analysis revealed the mutation to be clearly linked to haplotype III allele inherited from the proband's father. These results indicate this novel Serine 25 mutation originated in the paternal germline mosaicism. It is possible that the vaccination had an influence on the unique clinical picture, but this remains uncertain. PMID- 11870695 TI - Congenital inflammatory myopathy: a demonstrative case and proposed diagnostic classification. AB - There have been few reports of congenital inflammatory myopathy in the literature, and most of these have been associated with congenital muscular dystrophy. We review the literature and present a case with electromyographic and muscle biopsy evidence of congenital inflammatory myopathy with onset in the perinatal period and no evidence of a congenital muscular dystrophy. There was evidence of subjective improvement following corticosteroid administration (approximately 1 mg/kg per day) with a concomitant normalization of the serum creatine kinase activity. Of particular interest in the case was the history of maternal infection, suggesting a possible postinfectious molecular mimicry as the etiology of the muscle inflammation. This case suggests that a rare form of congenital inflammatory myopathy does exist that is not associated with a congenital muscular dystrophy. A preliminary classification scheme is proposed to separate true congenital cases from those acquired after birth and those cases associated with congenital dystrophy. PMID- 11870697 TI - Electrical stimulation factors in potentiation of human quadriceps femoris. AB - Potentiation is the enhancement of force seen after repetitive activation of skeletal muscle. The frequency and duration of stimulation, total number of pulses delivered to the muscle, and the peak forces or force-time integrals produced by the stimulation all have been suggested to affect the degree of potentiation. The purpose of this study was to determine the effect of the electrical stimulation characteristics on the development of post-activation potentiation. Eleven subjects were tested with five potentiating trains, including 12-pulse 100-HZ, 31-HZ, 14-HZ, and 5-HZ, and 6-pulse 14-HZ trains. The potentiating trains differed in stimulation frequency, train duration, and total number of pulses. They also produced different peak forces and force time integrals from the activated muscles. Our results showed that the 12-pulse 5-HZ train produced about 50% less potentiation than the other four potentiating trains. At stimulation frequencies of 14 HZ or higher, the total number of pulses delivered to the muscle was the primary factor in potentiation development. Furthermore, peak force and force-time integral had no effects on the rate or amount of potentiation. These results should help clinicians and researchers to design protocols that control for the effects of muscle potentiation. PMID- 11870696 TI - Spasticity causes a fundamental rearrangement of muscle-joint interaction. AB - Sarcomere length was measured in flexor carpi ulnaris (FCU) muscles from patients with severely spastic wrist flexion contractures (n = 6), as well as from patients with radial nerve injury and a normally innervated FCU (n = 12). Spastic FCU muscles had extremely long sarcomere lengths with the wrist fully flexed (3.48 +/- 0.44 microm) compared to the FCU muscles of patients with radial nerve injury (2.41 +/- 0.31 microm). In three of the patients with spastic wrist flexion contractures, the slope of the FCU sarcomere length-joint angle relationship was measured and found to be, essentially, normal (0.017 +/- 0.005 microm/degree, n = 3) suggesting that serial sarcomere number (and therefore muscle fiber length) was unchanged in spite of the dramatic absolute sarcomere length change. These results indicate that spasticity results in a major alteration of normal muscle-joint anatomical relationships that has not previously been recognized to our knowledge. We hypothesize that the results are explained either by the inability of muscle fibers to add serial sarcomeres in response to growth, or the selective loss of FCU muscle length secondary to the central nervous system lesion. PMID- 11870698 TI - New cyclooxygenase-2 inhibitors for treatment of experimental autoimmune neuritis. AB - We analyzed two new cyclooxygenase-2 (COX-2) inhibitors, celecoxib (SC-58635) and meloxicam, for the treatment of experimental autoimmune neuritis (EAN) in rats. Celecoxib and meloxicam significantly reduced clinical EAN score and histopathological damage of the sciatic nerve. They induced no serious side effects, whereas indomethacin used as a control caused severe intestinal ulceration and dysfunction of liver and kidney. These findings suggest that the new COX-2 inhibitors may be useful as additional therapeutic agents for patients with Guillain-Barre syndrome and chronic inflammatory demyelinating polyneuropathy. PMID- 11870699 TI - Acute rhabdomyolysis and brachial plexopathy following alcohol ingestion. AB - An unusual case of brachial plexopathy following alcohol-induced rhabdomyolysis is presented. The patient's rhabdomyolysis developed during sleep after an acute alcohol binge and there was no history of muscle trauma. It is thought that the brachial plexopathy developed due to direct compression of the plexus from swollen muscles of the shoulder girdle. The lack of similar reported cases despite the common clinical scenario of prolonged unconsciousness following excess alcohol intake suggests that other factors may be important in the development of muscle and nerve damage in susceptible individuals. PMID- 11870700 TI - Mycophenolate mofetil in the therapy of polymyositis associated with a polyautoimmune syndrome. AB - Mycophenolate mofetil 1.5 g daily (30 mg/kg body weight) was given to a patient with ankylosing spondylitis, ulcerative colitis, and severe refractory polymyositis after conventional treatment regimes had failed. No severe side effects occurred. Considerable improvement of clinical symptoms and electromyographic findings were seen within 6 months after the initiation of mycophenolate mofetil, allowing for tapering and discontinuation of methylprednisolone. Mycophenolate mofetil may be considered as an useful alternative in the treatment of polymyositis when standard therapeutic regimens fail. PMID- 11870701 TI - Titin and nebulin content in human skeletal muscle following eccentric resistance exercise. AB - We measured titin and nebulin content in muscle biopsies from the vastus lateralis before and 24 h after one bout of high-intensity eccentric knee extensor resistance exercise in seven men (26 +/- 3 years). Titin and nebulin content were significantly (P < 0.05) reduced after exercise by 30 and 15%, respectively. These results suggest that the structural components of the myofibrillar apparatus are degraded following high-intensity eccentric resistance exercise in humans. Loss of these proteins may have important implications for the mechanisms regulating the adaptive response of skeletal muscle to resistance exercise. PMID- 11870702 TI - Concomitant dermatomyositis and myasthenia gravis presenting with respiratory insufficiency. AB - We report a 28-year-old woman with a history of chronic immune-mediated hepatitis, in whom the simultaneous manifestation of dermatomyositis and myasthenia gravis resulted in severe neck extensor weakness and subacute respiratory insufficiency, followed by proximal muscle weakness and external ophthalmoplegia. Radiological signs of a thymoma were absent. The distinguishing clinical, electrophysiological, and biopsy findings are discussed. We suggest that an underlying immunoregulatory disorder was present, explaining the occurrence of three rare immune-mediated diseases in one patient. PMID- 11870703 TI - Unilateral cranial and phrenic nerve involvement in axonal Guillain-Barre syndrome. AB - A 49-year-old woman developed acute left facial, hypoglossal, and phrenic nerve palsies, as well as dysphagia and weakness in the neck and arms. Electrophysiologic studies showed an acute motor axonal neuropathy. Serum anti GM1 IgG antibody was positive. Intravenous immunoglobulin treatment resulted in good clinical recovery. The present report indicates that the cranial and phrenic nerves may be affected unilaterally in Guillain-Barre syndrome, and that there is clinical variability in the axonal subtype of this syndrome. PMID- 11870706 TI - Primary axonal degeneration in tarsal tunnel syndrome: fact or fiction? PMID- 11870708 TI - More than meets the eye: extraocular muscle is very distinct from extremity skeletal muscle. PMID- 11870704 TI - Acetylcholine receptors at the neuromuscular junction of regenerated muscle fibers. PMID- 11870709 TI - The utility of magnetic resonance imaging in evaluating peripheral nerve disorders. AB - The evaluation of peripheral nerve injuries has traditionally relied primarily on information gained from the clinical history, physical examination, and electrodiagnostic testing. Taken together, all of this clinical and diagnostic information often allows one to determine the location and severity of the underlying peripheral nerve problem. However, it may not be sufficient in diagnosing a focal entrapment neuropathy superimposed upon a more generalized peripheral neuropathy; localizing a focal lesion along a long segment of nerve which may be difficult to assess accurately with electrodiagnostic studies; distinguishing early between an axonotmetic grade of injury, which can recover through axonal regeneration, and a neurotmetic grade which cannot and therefore may benefit from a surgical exploration and repair procedure; and noninvasively diagnosing and determining the surgical resectability of peripheral nerve mass lesions such as tumors. The goal of this review is to illustrate how standard and evolving magnetic resonance imaging techniques can provide additional information in dealing with some of these problems. PMID- 11870710 TI - Rhabdomyolysis: a review. AB - Rhabdomyolysis, a syndrome of skeletal muscle breakdown with leakage of muscle contents, is frequently accompanied by myoglobinuria, and if sufficiently severe, acute renal failure with potentially life-threatening metabolic derangements may ensue. A diverse spectrum of inherited and acquired disorders affecting muscle membranes, membrane ion channels, and muscle energy supply causes rhabdomyolysis. Common final pathophysiological mechanisms among these causes of rhabdomyolysis include an uncontrolled rise in free intracellular calcium and activation of calcium-dependent proteases, which lead to destruction of myofibrils and lysosomal digestion of muscle fiber contents. Recent advances in molecular genetics and muscle enzyme histochemistry may enable a specific metabolic diagnosis in many patients with idiopathic recurrent rhabdomyolysis. PMID- 11870711 TI - Continuous myonuclear addition to single extraocular myofibers in uninjured adult rabbits. AB - Extraocular muscles (EOM) are unique among mammalian skeletal muscles in that they normally express molecules associated with muscle development and regeneration. In this study we show that satellite cells of EOM, unlike those of other skeletal muscles, continually divide in the normal, uninjured adult. Adult EOM contained activated satellite cells positive for the myogenic regulatory factor MyoD. EOM satellite cells did not require a prolonged activation period prior to onset of cell division and differentiation in vitro. EOM satellite cells incorporated bromodeoxyuridine (brdU), a marker for cell division, and with longer postlabeling survival, brdU-labeled nuclei populated EOM myofibers. This was not seen with leg muscle. These findings suggest the possibility that continual division of satellite cells and fusion of their daughter myocytes with existing adult EOM myofibers contribute to the unique sparing or susceptibility of EOM to certain muscle diseases. PMID- 11870712 TI - Effect of exercise on stability of chronically enlarged motor units. AB - Chronic denervation syndromes such as the post-polio syndrome are associated with progressive muscle weakness and fatigue after motoneuron death. Neither the etiology nor the management of these syndromes is clear. To address this issue, we partially denervated rat hindlimb muscles for 1 or 12 months and examined whether chronically enlarged motor units (MUs) become destabilized with time and further destabilized by daily running on exercise wheels. MU enlargement, measured electrophysiologically and morphologically was significantly reduced at 12 months in extensively denervated muscles, and to a lesser extent in moderately denervated muscles, as compared to the findings at 1 month. A 1-month period of running exercise further reduced the size of the chronically enlarged MUs in the extensively denervated muscles. We have therefore (1) successfully established a rat model of time-related MU size reduction, in which destabilization of chronically enlarged MUs results in loss of axonal terminals, and (2) demonstrated that nonphysiological activity has small but significant effects of further destabilizing the chronically enlarged MUs. PMID- 11870713 TI - Psychometric evaluation of a new handicap scale in immune-mediated polyneuropathies. AB - A new handicap measure, the Rotterdam nine-item handicap scale, was developed and its validity, reliability, and responsiveness evaluated in patients with immune mediated polyneuropathies. We evaluated 113 stable patients, of whom 83 had Guillain--Barre syndrome (GBS), 22 had chronic inflammatory demyelinating polyneuropathy (CIDP), and 8 had a gammopathy-related polyneuropathy. We also studied 20 patients with recently diagnosed GBS (n = 7) or CIDP (n = 13) and changing clinical conditions (longitudinal group). Significant discriminatory validity and correlation with the Rankin scale were demonstrated for the Rotterdam nine-item handicap scale (stable group: Spearman's test, r = minus sign.76 to minus sign.78; longitudinal group: intraclass correlation coefficient, r =.83; P <.0001). Also, good reliability (r =.89--.98; P <.0001) and high responsiveness values (standardized response mean values >.8) were obtained for the Rotterdam nine-item handicap scale. In contrast to the Rankin scale, the Rotterdam scale not only provided information regarding mobility but also highlighted physical independence, occupation, and social integration. These results illustrate the clinical usefulness of the Rotterdam nine-item handicap scale under these conditions. PMID- 11870715 TI - Lipofuscin accumulation in the vastus lateralis muscle in patients with chronic obstructive pulmonary disease. AB - Exercise-induced oxidative stress has been reported in patients with chronic obstructive pulmonary disease (COPD) and may play a role in muscle fatigue. It is speculated that oxidative stress during exercise originates from the contracting muscles but this has not been documented. The accumulation of lipofuscin, a marker of cellular oxidative damage, was evaluated in the vastus lateralis muscle in 17 patients with COPD and 10 healthy subjects of similar age. Each subject performed a stepwise exercise test up to maximal capacity during which oxygen uptake (VO(2)) was measured. Resting and peak exercise blood gases were also obtained. Two indices of lipofuscin accumulation were used: lipofuscin inclusions/fiber ratio (LI/F) and lipofuscin inclusions/fiber cross-sectional area ratio (LI/CSA). These ratios were also determined for each specific fiber type. LI/F (P < 0.01) and LI/CSA (P < 0.01) were greater in COPD compared to healthy subjects. LI/F and LI/CSA for all fiber types were also greater in COPD (P < 0.001). In both groups, LI/F (P < 0.001) and LI/CSA (P < 0.01) were higher in type I than in type II fibers. LI/F and LI/CSA did not correlate significantly with resting PaO(2) and SaO(2), peak VO(2), and DeltaPaO(2) and DeltaSaO(2) during exercise (P > 0.05). Increased lipofuscin accumulation, a marker of oxidative damage, was found in the vastus lateralis muscle in patients with COPD compared to healthy subjects. Oxidative damage of muscle tissue may thus be involved in skeletal muscle dysfunction and wasting in COPD. PMID- 11870716 TI - Localized bioimpedance analysis in the evaluation of neuromuscular disease. AB - Localized bioimpedance analysis is a novel, noninvasive technique with potential application to neuromuscular disease. In this procedure, high-frequency alternating current is passed through muscle, and parameters related to the consequent voltage pattern are evaluated. Currents flowing perpendicular to muscle fibers encounter many more cell membranes than do currents flowing parallel to them, producing surface voltage patterns that are altered by disease. Using this technique, 45 normal subjects and 25 patients with various neuromuscular diseases were studied, including 4 with amyotrophic lateral sclerosis, 4 with inflammatory myopathy, and 11 with inclusion-body myositis. Two parameters, the spatially averaged phase and the effective longitudinal resistivity, were altered in patients with neuromuscular disease. Reductions in phase correlated to disease progression, whereas normalization of phase correlated with disease remission. In patients with inclusion-body myositis, a unique pattern of reduced phase and elevated resistivity was identified. These findings suggest that localized bioimpedance analysis has the potential of playing a substantial role in the diagnostic and therapeutic evaluation of neuromuscular disease. PMID- 11870714 TI - Motor conduction studies in Miller Fisher syndrome with severe tetraparesis. AB - Some patients with Miller Fisher syndrome (MFS) have a severe tetraparesis such as that observed in Guillain--Barre syndrome (GBS). To determine whether pathophysiologic differences exist between the tetraparesis in MFS and that in GBS, we compared clinical and motor conduction findings in 4 MFS patients who developed severe tetraparesis with those in 5 MFS patients without tetraparesis, and 14 GBS patients. MFS patients with or without tetraplegia had normal motor conduction velocities, distal motor latencies, compound muscle action potential (CMAP) amplitudes, and F-wave latencies. CMAP amplitude tended to be lower in tetraparetic MFS patients than in MFS patients without tetraparesis, but not significantly. F-wave occurrence was slightly reduced in 1 MFS patient with tetraparesis and 1 MFS patient without tetraparesis. Motor conduction parameters were abnormal in 13 of 14 patients with GBS, and showed demyelinating features in 10. Our results suggest that the pathophysiology of tetraparesis in MFS differs from that in GBS. PMID- 11870717 TI - Diagnosis of dystrophinopathy by skin biopsy. AB - We studied the expression of dystrophin in skin biopsy samples from 19 patients with neuromuscular diseases. Immunohistochemical procedures for dystrophin analyses were performed using monoclonal antibodies for three different domains. Arrector pili muscles, which are smooth muscles in the skin, expressed dystrophin in the patients with limb-girdle muscular dystrophy (5), facioscapulohumeral muscular dystrophy (1), and spinal muscular atrophy (3), and in normal controls (2). The C-terminus of dystrophin was slightly expressed in the patients with Duchenne muscular dystrophy, whereas the rod domain and N-terminus were absent. In one patient with Becker muscular dystrophy, the expression of dystrophin was reduced. The mosaic of dystrophin positive and negative smooth muscle fibers was observed in a manifesting carrier of Duchenne muscular dystrophy. Our results suggest that skin biopsy is very useful for the diagnosis of Duchenne/Becker muscular dystrophy and manifesting carrier of Duchenne muscular dystrophy, and can be performed even at an advanced stage of the disease. PMID- 11870718 TI - Subtle involvement of the sympathetic nervous system in amyotrophic lateral sclerosis. AB - The literature on the involvement of the autonomic nervous system (ANS) in amyotrophic lateral sclerosis (ALS) is conflicting. We therefore investigated several aspects of autonomic function, namely muscle sympathetic nerve activity (MSNA), blood pressure, cardiac function (electrocardiogram; ECG), and respiration in 16 patients with sporadic ALS and in 12 age-matched healthy volunteers, both at rest and during sympathoexcitatory stimulation. We measured MSNA by provoking venous pooling during short-lasting lower body negative pressure (LBNP) and during the cold pressor test (CPT). To assess the vagal (baroreflex) control of heart rate (HR), we measured spontaneous baroreflex sensitivity (BRS). To assess the involvement of the ANS beyond the cardiovascular system, we measured the sympathetic skin response (SSR). The stand-up test showed that none of the subjects had orthostatic intolerance. In comparison with the control group, the ALS patients had an increased HR and a decreased BRS at rest, and a reduced MSNA response to LBNP. The CPT response was normal and the total MSNA at rest did not differ significantly from that of controls. The latencies of the palmar and plantar SSR were prolonged, and in 3 ALS patients there was no plantar SSR. The results indicate that the sympathetic nervous system shows subtle abnormalities in ALS, predominantly sympathetic overactivity. They also point to the involvement of the preganglionic sympathetic column as the cause of the higher sympathetic activity and the absence of SSR. The higher sympathetic activity is postulated to be due to changes in modulation of the sympathetic system, whereas the absence of the SSR is probably caused by disruption of the reflex pathway. PMID- 11870719 TI - Adhesion molecule expression in experimental myositis. AB - Experimental allergic myositis (EAM) in Lewis rats, induced with partially purified myosin, is regarded as a model of human polymyositis. To clarify the role of adhesion molecules in the pathogenesis of EAM in Lewis rats, we investigated intramysial expressions of the intercellular adhesion molecule (ICAM)-1 and vascular cell adhesion molecule (VCAM)-1, and the serum level of soluble ICAM-1 in EAM rats. All the EAM rat muscles had scattered inflammatory foci, as well as cell infiltration and necrosis, by week 4 after the initial immunization (i.e., day 0 after the last immunization). As compared with the control muscles, ICAM-1 and VCAM-1 were strongly expressed immunohistochemically in the endothelium of vessels in the endomysium and perimysium, and to lesser extents in the inflammatory infiltrates and on the sarcolemma of nonnecrotic muscle fibers adjacent to the inflammatory infiltrates or invaded muscle fibers. ICAM-1 in the muscle extracts and sera from EAM rats increased on each test day, as compared with extracts from the normal controls. The values peaked on day 0 after the last immunization, then gradually decreased with time. ICAM-1 elevations in the muscle extracts were correlated with the percent of sections that had inflammatory lesions (P = 0.032) and the histological scores (P = 0.005) on day 0, whereas there was no significance on days 3 and 7. These findings suggest that the adhesion molecules ICAM-1 and VCAM-1 increase in the early stage of EAM, and function in the initiation of the inflammatory process of myositis. PMID- 11870721 TI - Qualitative and quantitative changes in acetylcholine receptor distribution at the neuromuscular junction following free muscle transfer. AB - The qualitative and quantitative changes in acetylcholine receptor distribution were studied in the gracilis muscle of the Wistar rat following free neurovascular transfer. Even at 30 weeks after transfer, the morphology of the neuromuscular junction failed to return to the presurgical state. The number of acetylcholine receptors at the reinnervated neuromuscular junction also remained lower than the control. The persistent weakness following free neurovascular muscle transfer may be attributed to these qualitative and quantitative changes at the neuromuscular junction. PMID- 11870720 TI - Metabolic costs of force generation for constant-frequency and catchlike-inducing electrical stimulation in human tibialis anterior muscle. AB - Metabolic costs of force generation were compared for constant-frequency and catchlike-inducing electrical stimulation. Repetitive catchlike-inducing trains consisted of 2 interpulse intervals (IPIs) at 12.5 ms, 1 IPI at 25 ms, and 5 IPIs at 50 ms. Constant-frequency trains consisted of 8 IPIs at 37.5 ms. One train was delivered to the peroneal nerve every 2.5 s for 36 times under ischemic conditions. Anaerobic adenosine triphosphate (ATP) turnover was determined using 31-phosphorus magnetic resonance spectroscopy (P-MRS) of the human tibialis anterior muscle. Compared with constant-frequency trains, catchlike-inducing trains produced a faster force generation and were more effective in maintaining the force--time integral as well as peak force. However, ATP costs of force generation were similar for the catchlike-inducing and constant-frequency stimulation (6.7 plus/minus 1.1 and 6.6 plus/minus 1.0 micromol ATP/kg wet weight/Ncenter dots, respectively, P = 0.601). This suggests that the positive effects of catchlike-inducing stimulation on force maintenance are mediated by potentiated Ca(2+) release from the sarcoplasmic reticulum rather than by lower metabolic costs of muscle force generation. Our findings also suggest that catchlike-inducing stimulation produces larger forces in fatigued muscle than constant-frequency trains and thus may be beneficial for muscle training or rehabilitation when muscle loading needs to be maintained in repetitive contractions. PMID- 11870722 TI - Block of recombinant nicotinic receptor channels by IgG from myasthenic patients. AB - In myasthenia gravis (MG), specific antibodies directed against nicotinic acetylcholine receptor (nAChR) channels are produced. Recently, a distinct blockade of native embryonic-type nAChR channels by purified IgG from MG patients has been demonstrated. In the present study, we investigated the interaction of myasthenic IgG with recombinant embryonic- or adult-type nAChR channels expressed on human embryonic kidney (HEK) 293 cells. For the experiments, the patch clamp technique was applied in combination with an ultra-fast application system for agonists. Repetitive 20-ms pulses of 1 mM acetylcholine (ACh) were applied with or without purified IgG added to the perfusion system. The purified IgG from MG patients blocked currents through both, embryonic- and adult-type nAChR channels to a similar extent. A block by myasthenic IgG of adult-type nAChR channels, which are present exclusively at the postsynaptic membrane of the neuromuscular synapse, was shown to occur. The reversible pharmacological blockade of adult type nAChR channels by myasthenic IgG may be one factor responsible for rapid fluctuations of weakness or rapid recovery of muscle strength after plasmapheresis in MG. PMID- 11870723 TI - Comparison of potentiated and unpotentiated twitches as an index of muscle fatigue. AB - Recent data suggest that the potentiated twitch is a more sensitive index of contractile fatigue than is the unpotentiated twitch. We hypothesized that after a potentially fatiguing load, the fall in twitch amplitude of the potentiated twitch would be significantly greater than that of an unpotentiated twitch. We compared the response of the potentiated and unpotentiated twitches to a series of potentially fatiguing loads using magnetic stimulation of the femoral nerve in 10 healthy subjects. The baseline unpotentiated quadriceps twitch force (TwQu), potentiated quadriceps twitch force (TwQp), and maximal voluntary contraction (MVC) were 129 plus minus 6 N, 198 plus minus 6 N, and 622 plus minus 25 N, respectively. During a fatigue protocol that was designed to induce a spectrum of fatigue from mild to marked, the percent fall in quadriceps twitch force was significantly greater for the potentiated method than for the unpotentiated method at all levels of fatigue (P <.005). The within-subject within-day coefficient of variation was 7.5 plus minus 0.5% for TwQu and 5.6 plus minus 0.9% for TwQp. Thus, TwQp is reproducible and is superior to TwQu for detecting early muscle fatigue. PMID- 11870724 TI - Motor unit number estimation in infants and children with spinal muscular atrophy. AB - Spinal muscular atrophy (SMA) is a disease of lower motor neurons. Motor unit number estimation (MUNE) is an electrophysiologic method to estimate the number of motor neurons innervating a muscle group. We applied the multiple point stimulation technique to the ulnar nerve--hypothenar muscle group to study lower motor neuron loss in 14 SMA subjects, including those presymptomatic, and varying from newborn through 45 years of age. Preliminary data support the value of MUNE to help understand the time course of motor neuron loss in SMA. PMID- 11870725 TI - Postexercise facilitation of motor evoked potentials following transcranial magnetic stimulation: a study in normal subjects. AB - The size of the motor evoked potential (MEP) elicited by transcranial magnetic stimulation increases soon after a nonexhaustive voluntary contraction of the target muscle (postexercise facilitation). Our aim was to determine whether the duration or intensity of voluntary muscle contraction influenced postexercise facilitation in normal subjects. We recorded the MEP from the thenar muscles following contractions of different durations (5, 15, and 30 s) and intensities (10%, 25%, and 50% of maximal voluntary contraction). We found that every combination of the tested intensities and durations of physical effort could induce postexercise MEP facilitation. Although the degree of postexercise MEP facilitation was comparable across the different durations and intensities, the maximal facilitation was observed with the shortest and strongest muscle contraction. Our study thus defines the optimal setting to study postexercise facilitation for clinical purposes. PMID- 11870726 TI - Hypokalemic periodic paralysis associated with malignant hyperthermia. AB - Hypokalemic periodic paralysis is in most cases related to mutations within the dihydropyridine receptor gene. Susceptibility to malignant hyperthermia has been linked to a different part of the same gene, but is more frequently caused by mutations within the ryanodine receptor gene. We report the association of the two disorders in a patient for whom the most frequent mutations for hypokalemic periodic paralysis were not found. This suggests further genetic heterogeneity of this condition, the interest of this case residing in the known coupling between dihydropyridine and ryanodine receptors. PMID- 11870727 TI - Transient forearm conduction block in the median nerve. AB - We present two cases referred for electrophysiological confirmation of carpal tunnel syndrome (CTS). Initial nerve conduction studies were normal. Approximately 20 min into the examination, both patients developed sensory symptoms and weakness in the distal median nerve territory while the elbow was extended and forearm supinated. Further studies demonstrated complete conduction block across the forearm in the median motor and sensory nerve fibers. When measurable, conduction velocities remained normal or were modestly slow. Complete clinical and electrophysiological recovery occurred within 2 min following forearm pronation, suggesting that dysfunction was probably due to focal transient ischemia. Patients describing increased sensory symptoms during routine electrophysiological assessments for CTS should be investigated to rule out the possibility of a more proximal abnormality. PMID- 11870728 TI - Chronic eosinophilic perimyositis with persistent myalgias. AB - A 43-year-old man with an 8-year history of hypereosinophilia was evaluated for persistent muscle pain. Methotrexate and corticosteroids were ineffective. Examination, limited by pain even with passive motion, showed only mild weakness. Electromyography and muscle enzymes were normal. A needle muscle biopsy specimen revealed eosinophilic perimyositis. This case illustrates that the diagnosis of eosinophilic perimyositis requires histopathological evaluation, which should be pursued in patients with eosinophilia and persistent myalgia despite normal electromyography and muscle enzymes. PMID- 11870729 TI - Quality of life in patients with myasthenia gravis. PMID- 11870731 TI - Severe backache in Guillain-Barre syndrome. PMID- 11870735 TI - Capillary electrophoresis of anionic analytes in methanol: effect of counter-ions on electrophoretic mobility. AB - Mobilities of 11 substituted benzoates and 3 nitrophenolates were determined in non-aqueous methanol with Li+, Na+, K+, Rb+, and tetrabutylammonium (Bu4N+) as counter-ions of the background electrolyte. The influence of the ionic concentration of the background electrolyte on the mobility of the analyte anions is more pronounced compared to aqueous solutions. The deviation from the dependence of the mobilities on the ionic strength from the Debye-Huckel-Onsager theory indicates the occurrence of ion-pair formation. For a given ion concentration (10 mmol/L), the decrease of the analyte mobility follows the counter-ion sequence Li+ < Na+ < K+ < Rb+, which is the inverse order of their Stokes radii. Bu4N+ as counter-ion has a similar effect on the analyte mobility than Li+ (which has the same Stokes radius, but a six times smaller crystal radius). Exceptions are some di- and trihydroxybenzoates. The mobilities in methanol and in water with the same counter-ion (Na+) at a given ionic concentration show very low correlation. PMID- 11870736 TI - Electrophoretic mobilities of large organic ions in nonaqueous solvents: determination by capillary electrophoresis in propylene carbonate, N,N dimethylformamide, N,N,-dimethylacetamide, acetonitrile and methanol. AB - The mobilities of the monocharged permanent tertraphenylphosphonium cation and tetraphenylborate anion are determined by capillary zone electrophoresis in different organic solvents as a function of the ionic strength, I, of the background electrolyte. The nonaqueous solvents are propylene carbonate (PC), N,N dimethylformamide (DMF), N,N,-dimethylacetamide (DMA), acetonitrile (MeCN) and methanol (MeOH). The ionic strength is between 5 and 50 mmol/L. The mobility as a function of I is in good agreement with the theory of Debye, Huckel and Onsager (DHO), extended by the ion size parameter as introduced by Falkenhagen and Pitts. The values of the limiting DHO slopes of the mobility vs. I curves (the slopes express the influence of the solvent on the reduction of the mobility with increase of I) decrease in the order MeCN > MeOH > DMF > DMA > PC. Absolute mobilities (obtained by extrapolation to I = 0) of a particular ion differ by a factor of about 7 between the solvents. However, constancy within 10% is observed for their Walden products (the absolute mobility multiplied with the solvent's macroviscosity). The role of dielectric friction on the mobility of the present monocharged, large analyte ions is discussed according to the theory of Hubbard and Onsager. Based on the radii of the ions, the static permittivity of the solvent and its permittivity at infinite frequency, and the relaxation time of polarization, an equal contribution of dielectric and hydrodynamic friction is predicted in MeOH as solvent. Experimental data are in contrast to this prediction, indicating the overestimation of dielectric friction, and the dominance of hydrodynamic friction on the migration of the analyte ions in all solvents under consideration. PMID- 11870737 TI - Dielectric friction as a mechanism for selectivity alteration in capillary electrophoresis using acetonitrile-water media. AB - The mobilities of a series of aromatic carboxylates and sulfonates, ranging in charge from -1 to -4, were investigated as a function of acetonitrile concentration in the electrophoretic buffer. Absolute mobilities were determined by extrapolation of the effective mobilities to zero ionic strength according to the Pitts' equation. In general, anions of higher charge were more strongly influenced by ionic strength, with similarly charged anions experiencing ionic strength effects that were not significantly different at the 95% confidence level. Furthermore, the relative magnitudes of the Onsager slopes varied with acetonitrile content according to the z/(etaepsilon(1/2)) dependence in the electrophoretic effect of the Pitt's equation. Addition of acetonitrile to the electrophoretic media resulted in changes in the absolute mobilities of the anions. These acetonitrile-induced selectivity alterations were attributed to dielectric friction. As predicted by the Hubbard-Onsager model of dielectric friction, changes in sulfonate mobility were shown to correlate to changes in solvent viscosity (eta), dielectric constant (epsilon), and relaxation time (tau). The combined effects of ionic strength and dielectric friction caused analytes with higher charge-to-size ratios to be slowed to a greater extent upon addition of acetonitrile compared to those with lower charge-to-size. For example, at 75% acetonitrile and 20 mM ionic strength, a migration order reversal occurred between the triply and singly charged sulfonates. PMID- 11870738 TI - Nonaqueous capillary electrophoresis with alcoholic background electrolytes: separation efficiency under high electrical field strengths. AB - The effect of high voltage on capillary electrophoresis (CE) separations of anionic analytes in nonaqueous separation media was investigated. Methanol, ethanol, 1-propanol, and 1-butanol were tested as background electrolyte (BGE) solvents. Experiments were carried out with a laboratory-built CE instrument suitable for high-voltage separations. Potentials up to 60 kV were applied with reversed polarity to generate unusually high field strengths (e.g. 2000 Vcm-1) and so achieve fast and efficient separations. Highest separation efficiencies were obtained with propanol as BGE solvent, and the dependency of the efficiency on the separation voltage was more or less linear. With the other alcohols, separation efficiency decreased or remained roughly constant with increasing absolute voltage. The separation efficiencies are discussed in terms of longitudinal diffusion, Joule heating, and analyte interaction with the capillary wall. Capillary preconditioning had a varied effect on the separations in the different BGEs as the BGE and the conditioning process affected the electroosmotic flow (EOF) velocity and direction. PMID- 11870739 TI - Special aspects of detection methodology in nonaqueous capillary electrophoresis. AB - Over the recent years considerable efforts have been directed to the design of powerful detector arrangements for capillary electrophoresis (CE). The analytical characteristics of the detector have a great influence on the overall analytical performance of CE investigations. The major detection methods in CE, such as UV Vis absorbance, fluorescence, mass spectrometry and electrochemical detection, have successfully been adapted also to nonaqueous capillary electrophoresis (NACE). However, the different properties of organic solvent systems require some modification of detector concepts and design compared to aqueous CE. The advances of detector development and application in NACE are reported and discussed with emphasis on methodical aspects. PMID- 11870740 TI - Field-amplified sample stacking and nonaqueous capillary electrophoresis determination of complex mixtures of polar aromatic sulfonates. AB - Nonaqueous CE and field-amplified sample stacking have been used in the determination of complex mixtures of polar aromatic sulfonates (AS; mainly benzene- and naphthalenesulfonates) of environmental concern. The analytical procedure consists of an on-column aqueous sample enrichment, followed by the nonaqueous electrophoretic determination of stacked aromatic sulfonates. Various organic solvents were used as separation medium, acetonitrile and N methylformamide gave the best results. Optimum capillary electrophoresis separation is obtained with ammonium acetate (25 mM) dissolved in N methylformamide-methanol (90:10) as background electrolyte. This combined method was applied to the analysis of surface water samples spiked with selected aromatic sulfonates derivatives. PMID- 11870741 TI - Comparison of aqueous and nonaqueous carrier electrolytes for the separation of penicillin V and related substances by capillary electrophoresis with UV and mass spectrometric detection. AB - A method for the determination of penicillin V together with its impurities and by-products formed during biosynthesis, using capillary electrophoresis (CE) with UV and electrospray-mass spectrometric (ESI-MS) detection is presented. Aqueous and nonaqueous electrolytes containing 20 mM ammonium acetate were investigated to determine their suitability for the separation of these analytes. These carrier electrolytes were optimized with respect to the pH and the solvent/s used (water, methanol, acetonitrile, ethanol and isopropanol) and it was shown that although the nonaqueous electrolytes offered unique separation selectivities, the best results in terms of selectivity and sensitivity were obtained for the aqueous system. Finally, the applicability of this method for the analysis of a mixture representative of a real fermentation broth was demonstrated using an aqueous carrier electrolyte with both UV and ESI-MS detection. PMID- 11870742 TI - Comparison of three different anionic surfactants for the separation of hydrophobic compounds by nonaqueous capillary electrophoresis. AB - The effect of the three different surfactants, sodium dodecyl sulfate (SDS), diethylhexyl sodium sulfosuccinate (AOT), and taurodexycholic acid sodium salt (STDC) on the nonaqueous capillary electrophoretic separations of hydrophobic compounds were compared with formamide containing 20 mM K2HPO4 as electrolyte solvent. Separations of all selected uncharged hydrophobic compounds, e.g., p arylacetophones were shown to be strongly dependent on the kind of surfactant. The electrolyte containing 180 mM SDS provided the best result for the selected samples. PMID- 11870743 TI - Ionic liquids as electrolytes for nonaqueous capillary electrophoresis. AB - Acetonitrile is a well-suited medium for nonaqueous capillary electroseparations and enables extending the range of applications of capillary electrophoresis (CE) techniques to more hydrophobic species. In this study, the dialkylimidazolium based low temperature melting organic salts know as "ionic liquids" (ILs) are used as electrolytes. At room temperature these liquids are miscible with acetonitrile which makes it easy to use them for adjustment of analyte mobility and separation. The anionic part as well as the concentration of an IL influence the general electrophoretic mobility of the buffer system. The separation of different analytes is achieved because they become charged in the presence of ILs in separation media. There is also a possibility for a complex formation between the solute and the electrolyte which alters the mobility of the solute. A selected application of separations of phenols and aromatic acids will be discussed. PMID- 11870744 TI - Nonaqueous affinity capillary electrophoresis investigation of small molecule molecular recognition. AB - The conditional binding constants for a bis-guanidinium-like receptor and a series of dicarboxylate ligands have been determined in two buffer/solvent systems, namely 25 mM ammonium acetate/1% acetic acid in acetonitrile/methanol (7:3 v:v) and 30 mM N-methyl morpholine/15 mM methanesulfonic acid in acetonitrile/methanol (9:1 v:v). The latter buffer has not been applied before in capillary electrophoresis. The binding constants in both solvent systems decrease as the dicarboxylate length increases. The binding constants are larger in the less competitive N-methyl morpholine buffer. The dicarboxylates associate only weakly with a dicationic analog of the receptor, p-xylyl trimethylammonium, which is not a hydrogen bond donor. PMID- 11870745 TI - Interface for coupling nonaqueous wide-bore capillary electrophoresis with mass spectrometry. AB - A liquid-junction-type interface where a thin spraying capillary is inserted inside the separation capillary was constructed for coupling nonaqueous wide-bore capillary electrophoresis (CE) to mass spectrometry (MS). The robust structure of the interface provided fairly easy capillary handling. The study was carried out with uncoated CE capillaries of 200 and 320 microm inner diameter (ID). 1 Propanol-acetonitrile (80:20 v/v) with acetate electrolyte provided a low conducting medium for CE and good spraying conditions for electrospray ionization (ESI) without sheath-flow and drying gas. Methamphetamine, alprenolol, and levorphanol served as model compounds. Approximate detection limits with the 200 microm ID capillary were 35-265 ng/mL. PMID- 11870746 TI - Nonaqueous capillary electrophoresis-electrospray- mass spectrometry for the analysis of fluoxetine and its related compounds. AB - The potential of nonaqueous capillary electrophoresis was investigated for the simultaneous separation of fluoxetine hydrochloride, its meta-isomer, and other related compounds. The resolution of these compounds was compared in aqueous and nonaqueous media. Baseline separation of the studied solutes required a buffer electrolyte solution composed of 25 mM ammonium acetate and 1 M acetic acid in acetonitrile, an applied voltage of 30 kV and a temperature of 20 degrees C. Selectivity was considerably affected by the nature of the solvent (water, methanol, and acetonitrile). Moreover, substituting acetate by formate in the background electrolyte resulted in migration time changes, which were attributed to an ion-pairing phenomenon. Finally, the method was successfully coupled on line with electrospray ionization-mass spectrometry (ESI-MS) and allowed significant selectivity and sensitivity enhancement. The effect of ESI-MS parameters, such as nebulizing gas pressure, sheath liquid composition and flow rate, on resolution and method sensitivity was also discussed. PMID- 11870747 TI - Indirect fluorescence of aliphatic carboxylic acids in nonaqueous capillary electrophoresis using merocyanine 540. AB - A method for the analysis of aliphatic carboxylic acids (ACAs) in nonaqueous capillary electrophoresis (NACE) in conjunction with indirect laser-induced fluorescence (ILIF) using merocyanine 540 (MC 540) is described. Performing the analysis in organic solvent is advantageous when using MC 540, because of its greater quantum yield in aprotic solvent. To achieve a high dynamic reserve (DR) and optimize resolution, we have tested a number of aqueous mixtures containing alcohols and acetonitrile (ACN). The optimum buffer for the analysis of C2-C18 ACAs, in terms of sensitivity, resolution, and speed, is an aqueous mixture of 40% ACN, 30% ethanol, and 1 mM Tris at apparent pH 7.4 (adjusted with ascorbic acid). Under this condition, the DR is greater than 1000, thereby the limits of detection for acids are in the range of sub-microM to microM. Linear plots show that the dynamic ranges for the analysis of ACAs are at least two decades in concentration, with regression coefficients all greater than 0.98. The relative standard deviations of the migration times and peak heights for all ACAs are less than 2.0%. Furthermore, this simple and cost-effective method has been applied to the analysis of marine lipid concentrate, with the concentrations of 1.67+/-0.03 and 4.50+/-0.05 mM (n = 5) for C14 and C16 acids, respectively, in a tablet of marine lipid concentrate sample. PMID- 11870748 TI - Nonaqueous capillary electrophoretic separation of basic enantiomers using octakis(2,3-O-dimethyl-6-O-sulfo)-gamma-cyclodextrin, a new, single-isomer chiral resolving agent. AB - The enantiomers of 34 pharmaceutical weak-base analytes were separated by nonaqueous capillary electrophoresis in acidic methanol background electrolytes using the sodium salt of the new, single-isomer chiral resolving agent, octakis(2,3-O-dimethyl-6-O-sulfo)-gamma-cyclodextrin (ODMS). The effective mobilities, separation selectivities and peak resolution values of the weak-base analytes were determined as a function of the ODMS concentration in the 0-40 mM range and were found to follow the theoretical predictions of the charged resolving agent migration model (CHARM model) modified for ionic strength effects. Fast, efficient separations were achieved for both comparatively small and large enantiomers. PMID- 11870749 TI - Low-molecular-weight chiral cation exchangers: novel chiral stationary phases and their application for enantioseparation of chiral bases by nonaqueous capillary electrochromatography. AB - Cation exchange type chiral stationary phases (CSPs) based on 3,5-dichlorobenzoyl amino acid and amino phosphonic acid derivatives as chiral selectors (SOs) and silica as chromatographic support were developed and applied to enantiomer separations of chiral bases by nonaqueous capillary electrochromatography (NA CEC). As a rationale for efficient CSP development we adopted the combined use of the "reciprocity principle of chiral recognition" and nonaqueous ion-pair CE as screening assay. Thus, (S)-atenolol was employed as chiral counter-ion added to the BGE in CE and a series of N-derivatized amino acids and amino phosphonic acids were screened to derive reciprocally information on their chiral recognition abilities for atenolol enantiomers. Two SO candidates, namely N-(3,5 dichlorobenzoyl)-O-allyl-tyrosine and N-(4-allyloxy-3,5-dichlorobenzoyl)-1-amino 3-methylbutane phosphonic acid that have been identified as potential SOs in the CE screening were, after immobilization on thiol-modified silica, evaluated in cation-exchange NA-CEC. The strong chiral cation exchanger with the free phosphonic acid group exhibited enhanced enantioselectivity compared to the weak chiral cation exchanger with the carboxylic acid group. A wide variety of chiral bases could be successfully resolved on the strong chiral cation exchanger with alpha-values up to 2.2 and efficiencies up to 375000 m-1 including beta-blockers and other amino alcohols, local anesthetics like etidocaine, antimalarial agents like mefloquine, Troger's base, phenothiazines like promethazine, and antihistaminics. The influence of several experimental parameters (electrolyte concentration, acid-base ratio and acetonitrile-methanol ratio) was evaluated. PMID- 11870750 TI - Use of vancomycin silica stationary phase in packed capillary electrochromatography: III. enantiomeric separation of basic compounds with the polar organic mobile phase. AB - The separation of basic compounds into their enantiomers was achieved using capillary electrochromatography in 50 or 75 microm inner diameter (ID) fused silica capillaries packed with silica a stationary phase derivatized with vancomycin and mobile phases composed of mixtures of polar organic solvents containing 13 mM ammonium acetate. Enantiomer resolution, electroosmotic flow, and the number of theoretical plates were strongly influenced by the type and concentration of the organic solvent. Mobile phases composed of 13 mM ammonium acetate dissolved in mixtures of acetonitrile/methanol, ethanol, n-propanol, or isopropanol were tested and the highest enantioresolutions were achieved using the first mobile phase, allowing the separation of almost all investigated enantiomers (9 from 11 basic compounds). The use of capillaries with different ID (50 and 75 microm ID) packed with the same chiral stationary phase revealed that a higher number of theoretical plates and higher enantioresolution was achieved with the tube with lowest ID. PMID- 11870752 TI - Comparative study of separation and determination of triazines by micellar electrokinetic capillary chromatography and nonaqueous capillary electrophoresis: application to residue analysis in natural waters. AB - The separation and determination of a mixture of chloro- and methylthiotriazines in water samples by both micellar electrokinetic capillary chromatography (MEKC) and nonaqueous capillary zone electrophoresis (NA-CZE) were compared. The characteristics of both methods proved to be very similar in terms of separation efficiency and analysis times, but application of these methods for the analysis of triazines in natural waters, with a prior preconcentration step, revealed significant differences. A preconcentration step by solid-phase extraction (SPE) with Oasis HLB cartridges was accomplished for the determination of triazines at sub-ppb levels in drinking and river waters; when NA-CZE was used after this SPE step, electropherograms with fewer interferences and more stable baselines were obtained than when separation was carried out using MEKC. Another aspect related to the application to real samples was the lack of precision encountered upon evaluating the electrophoretic signals generated when using SPE coupled with NA CZE. Here, we demonstrate the importance of choosing an appropriate internal standard for analyte quantification. It is recommended that a triazine belonging to the same family as that of the triazine to be determined should be used as internal standard. PMID- 11870751 TI - Enantioseparations in nonaqueous capillary liquid chromatography and capillary electrochromatography using cellulose tris(3,5-dimethylphenylcarbamate) as chiral stationary phase. AB - The potential of the widely used chiral stationary phase for high-performance liquid chromatography (HPLC) enantioseparations, cellulose tris(3,5 dimethylphenylcarbamate) (CDMPC, sold under the trade name Chiralcel OD) was evaluated under the conditions of nonaqueous capillary electrochromatography (CEC). The effect of the particle size of the silica gel, the loading of CDMPC on the silica gel and nature of the organic solvent, as well as electrolyte salts on the separation characteristics were investigated. This study illustrates the applicability of CDMPC for obtaining highly efficient enantioseparations under the conditions of nonaqueous CEC. Comparative study of enantioseparations in capillary liquid chromatography (CLC) and CEC indicated the significant advantages of CEC such as higher plate number at the similar linear flow rates of the mobile phase as well as better tolerance of higher linear flow rates. PMID- 11870754 TI - Using nonaqueous capillary electrophoresis to analyze several quinolones in pig kidney samples. AB - We show the potential of nonaqueous capillary electrophoresis (NACE) for analyzing enrofloxacin (ENR), ciprofloxacin (CPR), danofloxacin (DAN), difloxacin (DIF), marbofloxacin (MAR), flumequine (FLU), and oxolinic acid (OXA) in pig kidney samples. We have studied the effects of parameters such as the composition of the organic media, the choice of electrolyte, the pH* of the background electrolyte (BGE), the addition of modifiers, and the reversal of electroosmotic flow. Separation was good with 20 mM ammonium acetate, 0.004% polycation hexadimethrine bromide (HDB), and 4% acetic acid (pH* 5.4) in methanol/acetonitrile (50:50 v/v) medium. We used a quick and simple sample preparation method, hydrochloric acid as an extractant and solid-phase extraction (SPE) with Baker C18 cartridges as the cleanup step. Recoveries for all quinolones were over 80%. PMID- 11870753 TI - Determination of tamoxifen and metabolites in mouse fetal tissue using nonaqueous capillary electrophoresis. AB - Tamoxifen (TAM), an antiestrogen, has been approved for use by women at risk for developing hormone-dependent breast cancer. Administration of TAM to pregnant CD 1 mice apparently results in reproductive tract toxicity in female offspring. However, there is little or no data describing potential TAM-induced fetal toxicity to women who may become pregnant while receiving prophylactic TAM treatment. In support of the National Toxicology Program's characterization of reproductive and developmental effects of TAM, the present work describes a capillary electrophoresis (CE)-based analytical technique used for detection of TAM and two major metabolites, N-desmethyltamoxifen (DMT), and 4-hydroxytamoxifen (4-HT) in CD-1 mouse fetal tissue. TAM-derived material was extracted from CD-1 mouse fetuses 2-12 h following TAM administration (100 mg/kg) to dams on gestation day 16. The presence of TAM, DMT, and 4-HT was confirmed in the solvent extracts by nonaqueous CE. The limit of detection of TAM by UV absorption was approximately 675 amol at a signal-to-noise ratio of 2:1. This work demonstrates both transplacental transport of TAM in CD-1 mice and a sensitive analytical technique for detecting low concentrations of TAM and similar compounds in biological tissues. PMID- 11870758 TI - Truly hypervariable DNA fingerprints due to exceptionally high mutation rates. AB - The power of DNA fingerprinting is due to comparatively high mutation rates of minisatellite and microsatellite DNA sequences. Studying the mating system of a parrot species (Burrowing Parrots, Cyanoliseus patagonus) using oligonucleotide probes, we observed mutation rates that are several orders of magnitude higher than those described anywhere in the literature. Most plausibly, the respective values are based on 3-4 loci with mutation rates of up to 100%. PMID- 11870757 TI - Activity staining of glutathione peroxidase after electrophoresis on native and sodium dodecyl sulfate polyacrylamide gels. AB - Glutathione peroxidase (GSH-Px), from commercial bovine erythrocytes or ammonium sulfate fractionations (30-45%, 45-60%, 60-75% and 75-90% saturations) of ginger rhizome, was detected on polyacrylamide gels after native polyacrylamide gel electrophoresis (PAGE) or sodium dodecyl sulfate (SDS)-PAGE. The gel was submerged in a 50 mM Tris-HCl buffer (pH 7.9) containing 13 mM glutathione and 0.004% hydrogen peroxide with gentle shaking for 10-20 min. The GSH-Px activity was stained with a solution containing 1.2 mM 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT) and 1.6 mM phenazine methosulfate (PMS) for 10 min. The clear zone of GSH-Px activity on a purple background was found in both native and SDS-PAGE gels. This fast and sensitive method can be used in the process of enzyme purification and characterization of mammalian or plant cells. PMID- 11870759 TI - Differential gene expression in response to copper in Acidithiobacillus ferrooxidans analyzed by RNA arbitrarily primed polymerase chain reaction. AB - Acidithiobacillus ferrooxidans is a chemoautotrophic bacterium that plays an important role in metal bioleaching processes. Despite the high level of tolerance to heavy metals shown by A. ferrooxidans, the genetic basis of copper resistance in this species remains unknown. We investigated the gene expression in response to copper in A. ferrooxidans LR using RNA arbitrarily primed polymerase chain reaction (RAP-PCR). One hundred and four differentially expressed genes were identified using eight arbitrary primers. Differential gene expression was confirmed by DNA slot blot hybridization, and approximately 70% of the RAP-PCR products were positive. The RAP-PCR products that presented the highest levels of induction or repression were cloned, sequenced and the sequences were compared with those in databases using the BLAST search algorithm. Seventeen sequences were obtained. The RAP-PCR product with the highest induction ratio showed similarity with the A. ferrooxidans cytochrome c. A high similarity with the thiamin biosynthesis gene thiC from Caulobacter crescentus was observed for another RAP-PCR product induced by copper. An RAP-PCR product repressed by copper showed significant similarity with the carboxysome operon that includes the ribulose-1,5-bisphosphate carboxylase/oxygenase complex from A. ferrooxidans and another copper-repressed product was significantly similar to the XyIN outer membrane protein from Pseudomonas putida. Finally, RAP-PCR products of unknown similarities were also present. PMID- 11870760 TI - Modification of capillary electrophoresis capillaries by poly(hydroxyethyl methacrylate), poly(diethylene glycol monomethacrylate) and poly(triethylene glycol monomethacrylate). AB - Modification of capillary electrophoresis (CE) capillaries by poly(hydroxyethyl methacrylate) (poly(HEMA), poly(diethylene glycol monomethacrylate) (poly(DEGMA) and poly(triethylene glycol monomethacrylate) (poly(TEGMA), was studied. Methods based on physical adsorption of the modifier and on its chemical binding were compared on the basis of the electroosmotic flow (EOF) reproducibility, the EOF dependence on the pH, the symmetry of the peak of positively charged tyramine, the stability of the coating and the separation of standard and milk proteins in the modified capillaries. Reproducible coatings were obtained by chemical binding of the polymers to the capillary walls and by coating with a solution of a polymer, as also demonstrated by the atomic force microscopy. PMID- 11870761 TI - Separation of DNA fragments in hydroxylated poly(dimethylacrylamide) copolymers. AB - The novel polymer matrices reported here are low-viscosity sieving media for DNA capillary electrophoresis. This new family of matrices comprises copolymers of N,N-dimethylacrylamide with different monomers which increase polymer hydrophilicity. All these new copolymers self-coat on fused-silica capillaries. Resolution, peak spacing and peak width were the parameters taken into account to assess the influence of polymer structure on separation selectivity and efficiency. This work demonstrates that the performance of polydimethylacrylamide (PDMA) can be improved through copolymerization with hydrophilic monomers. The improvement is related to the efficiency parameter. The new copolymers, due to their low viscosity high sieving capacity and ability to suppress EOF, represent a better alternative to PDMA and are suitable replaceable matrices for capillary and microchip electrophoresis. PMID- 11870762 TI - Whole-column imaging capillary electrophoresis of proteins with a short capillary. AB - Whole-column imaging capillary electrophoresis with a short capillary is discussed. A short capillary (3-6 cm) coated with either fluorocarbon or polyacrylamide was used as a separation capillary. The whole capillary was illuminated with 280 nm light, and the transmitted light was monitored by a linear charge-coupled device (CCD). For the short capillary, hydrodynamic flow caused by a subtle height difference between the anodic and cathodic reservoirs affected the sample migration in the capillary greatly. Several sample injection methods, including use of a cross connection, sealing of the capillary ends with a gel, and use of a gel-filled capillary, have been discussed. The experimental results showed that the peak height decreased and peak width increased with the electromigration distance. Therefore, higher sensitivity was obtained in a short capillary rather than a long capillary. The whole-column imaging CE with the short capillary has been applied for the study of conjugation reactions of protein cytochrome c with sodium dodecyl sulfate (SDS) and the dye Congo Red. The method has also been used for in situ monitoring of the electrophoretic protein desorption process. Our technique is a unique tool for the study of protein binding reactions and the interaction between analyte and inner wall of the capillary. PMID- 11870763 TI - Micellar electrokinetic chromatography with diode laser-induced fluorescence detection as a tool for investigating the fluorescence labeling of proteins. AB - We describe the use of micellar electrokinetic chromatography with diode laser induced fluorometry (MEKC-DLIF) as a tool to characterize a labeling reaction between sulfoindocyanine succinimidyl ester (Cy5), a cyanine fluorescent dye, and a model protein, bovine serum albumin (BSA). To decrease the influence of imprecise injection, methylene blue was added as an internal standard. Labeled BSA was completely separated from the unconjugated Cy5, and methylene blue under optimized conditions. A kinetic study of the reaction was performed by changing some parameters, such as reaction buffer pH, reaction buffer concentration and the initial concentration of BSA. A comparison between the current method and traditional methods was made. PMID- 11870764 TI - Highly sensitive chemiluminescence detection of copper(II) in capillary electrophoresis with field-amplified sample injection. AB - Field-amplified sample injection of copper(II) was investigated using capillary electrophoresis with chemiluminescence detection. The sensitivity of copper(II) has been improved markedly by the field-amplified sample injection technique and the detection limit reaches 2 x 10(-11) M. By injection of a short plug of water before sample introduction, the sensitivity can be further improved 5-fold and the detection limit reaches 4 x 10(-12) M. The relative standard deviations (n = 6) of the migration time and the peak height are 0.61% and 4.7% at 1.0 x 10(-9) M Cu(II), respectively. Parameters affecting the field-amplified sample injection, such as separation voltage and concentration of electrophoretic buffer, have been investigated. PMID- 11870765 TI - Analysis of amino acids by capillary zone electrophoresis with electrochemical detection. AB - The precapillary derivatization of 20 amino acids with naphthalene-2,3 dicarboxaldehyde (NDA) and CN(-) was investigated. All these derivatized amino acids could be oxidized on the carbon fiber microdisk bundle electrode except proline. Capillary zone electrophoresis with electrochemical detection was employed for the analysis of 19 amino acids. The optimum conditions of separation and detection were borate, pH 9.48, for the electrolyte, 18 kV for the separation voltage and 1.15 V versus a saturated calomel electrode for the detection potential. Limits of detection of concentration or mass for individual amino acids were between 1.7 x 10(-7) and 1.8 x 10(-6) mol/L or 84 and 893 amol (according to the signal-to-noise ratio of 3) for the injection voltage of 6 kV and injection time of 10 s. The relative standard deviations were between 0.80 and 2.3% for the migration times and 1.4 and 6.4% for the electrophoretic peak currents. From a mixture of 19 amino acids, 10 amino acids (Arg, Lys, Orn, Try, Ser, Ala, Gly, Cys, Glu, Asp) could be well separated. The other 9 amino acids appeared on three electrophoretic peaks. From the samples, in which the nine amino acids do not exist simultaneously, some of them could also be detected. The method was applied to the determination of amino acids in beer by the standard addition method. The recovery for the amino acids in beer was 91-109%. PMID- 11870766 TI - Capillary electrophoresis for detection of inherited disorders of purine and pyrimidine metabolism: a selective approach. AB - We developed a capillary electrophoresis method as a diagnostic tool for purine and pyrimidine metabolic disorders. Optimal conditions allowed the separation of the major diagnostic metabolites in urine samples within an analysis time of 10 min and with a separation efficiency of about 350,000 theoretical plates/m. The diagnostically important metabolites (adenine, adenosine, 2-deoxyadenosine, 2 deoxyguanosine, 2,8-dihydroxyadenine, guanosine, hypoxanthine, orotidine, orotic acid, and creatinine) were detectable at concentrations of 1.0-5.7 micromol/L. The method gives a linear calibration curve for tested purine and pyrimidine derivatives within the range of 5-500 micromol/L (r > 0.996) The coefficients of variation for the within- and between-day imprecisions were less than 3.2 and 5.8%, respectively. Characteristic abnormalities were detected in the electropherograms of urine samples from patients with purine and pyrimidine enzyme deficiencies. We provide electrophoretic and spectral characteristics of intermediates in purine and pyrimidine metabolism and possible artifacts from medication and their UV-absorbing compounds. Our method allows the detection of the majority of inborn errors of purine and pyrimidine metabolism. PMID- 11870768 TI - Application of micellar electrokinetic chromatography to the quality control of a pharmaceutical preparation containing three bronchodilators. AB - Theophylline(1,3-dimethylxanthine), dyphylline [7-(2,3 dihydroxypropyl)theophylline] and proxyphylline [7-(beta hydroxypropyl)theophylline] are three bronchodilators administered jointly in a single pharmaceutical preparation used against asthma. A micellar electrokinetic chromatography (MEKC) method for their resolution using a background electrolyte consisting of 20 mM tetraborate at pH 8.5 and 100 mM sodium dodecyl sulfate is proposed. The method was used to determine the three active principles in a pharmaceutical preparation. The small amount of sample required and the expeditiousness of the procedure allow content uniformity to be determined in individual tablets. The values of the validation parameters for the method (viz. selectivity, linearity, accuracy, precision, limit of detection, limit of quantitation and robustness) are reported. A complete factor design (2(3)x2) including pH, the surfactant concentration and the ionic strength of the background electrolyte as factors was used to estimate robustness. Based on the results, the method is robust enough for quantitation purposes. PMID- 11870767 TI - Partial filling micellar electrokinetic chromatography optimization studies of ibuprofen, codeine and degradation products, and coupling to mass spectrometry. AB - Studies have been performed to evaluate whether an on-line partial filling micellar electrokinetic chromatography (PF-MEKC) system could be applied to a recently developed MEKC method for the separation of ibuprofen, codeine and one of the degradation products. Attempts to couple the PF-MEKC system to MS have also been performed. SDS concentration, micellar zone length and concentration of acetonitrile in the buffer were optimized using factorial design. When a small micelle zone was injected directly after the sample introduction, the results improved markedly. The MS parameters have not been optimized, but the studies show promising results for the use of PF-MEKC-mass spectrometry for identification of the degradation products. PMID- 11870770 TI - Rapid microwell polymerase chain reaction with subsequent ultrathin-layer gel electrophoresis of DNA. AB - Large-scale genotyping, mapping and expression profiling require affordable, fully automated high-throughput devices enabling rapid, high-performance analysis using minute quantities of reagents. In this paper, we describe a new combination of microwell polymerase chain reaction (PCR) based DNA amplification technique with automated ultrathin-layer gel electrophoresis analysis of the resulting products. This technique decreases the reagent consumption (total reaction volume 0.75-1 microL), the time requirement of the PCR (15-20 min) and subsequent ultrathin-layer gel electrophoresis based fragment analysis (5 min) by automating the current manual procedure and reducing the human intervention using sample loading robots and computerized real time data analysis. Small aliquots (0.2 microL) of the submicroliter size PCR reaction were transferred onto loading membranes and analyzed by ultrathin-layer gel electrophoresis which is a novel, high-performance and automated microseparation technique. This system employs integrated scanning laser-induced fluorescence-avalanche photodiode detection and combines the advantages of conventional slab and capillary gel electrophoresis. Visualization of the DNA fragments was accomplished by "in migratio" complexation with ethidium bromide during the electrophoresis process also enabling real time imaging and data analysis. PMID- 11870769 TI - Analysis of arylamine isomers by micellar electrokinetic chromatography. AB - Analysis of carcinogenic substances is a high-priority area. Carcinogenic arylamines draw the analyst's attention because dyes and pigments are in production and used in large volumes. Identification of carcinogenic isomers of arylamines employing micellar electrokinetic chromatography (MEKC), a mode of capillary electrophoresis was studied as it offers better scope for separation science. Mixed micellar modes of MEKC techniques were employed to achieve acceptable analyses. Success of this analytical method was proved by real-sample analysis, which confirmed that this is a promising technique for the arylamine species. PMID- 11870771 TI - Towards disposable lab-on-a-chip: poly(methylmethacrylate) microchip electrophoresis device with electrochemical detection. AB - A fully disposable microanalytical device based on combination of poly(methylmethacrylate) (PMMA) capillary electrophoresis microchips and thick film electrochemical detector strips is described. Variables influencing the separation efficiency and amperometric response, including separation voltage or detection potential are assessed and optimized. The versatility, simplicity and low-cost advantages of the new design are coupled to an attractive analytical performance, with good precision (relative standard deviation RSD = 1.68% for n = 10). Applicability for assays of mixtures of hydrazine, phenolic compounds, and catecholamines is demonstrated. Such coupling of low-cost PMMA-based microchips with thick-film electrochemical detectors holds great promise for mass production of single-use micrototal analytical systems. PMID- 11870772 TI - Analysis of geometry effects on band spreading of microchip electrophoresis. AB - The geometry and the flow field conditions in the separation microchannel of an electrophoresis chip system may have important impact on the system's separation efficiency. Understanding the geometry effect on the flow field physics in the separation microchannel is beneficial to the design or operation of an electrophoresis system. The turns in a microfabricated separation microchannel generally results in degraded separation quality. To avoid this limitation, channels are constructed with different types of turns to determine the optimum design that minimizes turn-induced band broadening. We have designed and tested various geometric bend ratios to greatly reduce this so-called "racetrack" effect. The effects of the separation channel geometry, fluid velocity profile and bend ratio on the band distribution in the detection area are discussed. Results show that the folded square U-shaped channel is better for miniaturization and simplification. The band tilting was corrected and the racetrack effect reduced in the detection area when the bend ratio is 4:1. The detection time obtained from the present numerical solution matches very well with the experimental data. PMID- 11870773 TI - Quantification and evaluation of Joule heating in on-chip capillary electrophoresis. AB - We present the use of a novel, picoliter volume interferometer to measure, for the first time, the extent of Joule heating in chip-scale capillary electrophoresis (CE). The simple optical configuration for the on-chip interferometric backscatter detector (OCIBD) consists of an unfocused laser, an unaltered silica chip with a half-cylinder channel and a photodetector. Using OCIBD for millidegree-level noninvasive thermometry, temperature changes associated with Joule heating (2.81 degrees C above ambient) in on-chip CE have been observed in 90 microm wide and 40 microm deep separation channels. The temporal response of Joule heating in isotropically etched channels was exponential, with it taking an excess of 2.7 s to reach equilibrium. Buffer viscosity changes have also been derived from empirical on-chip thermometry data, allowing for the determination of diffusion coefficients for solutes when separated in heated buffers. In addition, OCIBD has allowed the reduction in separation efficiency to be estimated in the absence of laminar flow and due to increased molecular diffusion and lower buffer viscosity. A 7% reduction in separation efficiency was determined for a high current drawing buffer such as Tris-boric acid under an applied field of just 400 V/cm. Results indicate that heating effects in on-chip CE have been underestimated and there is a need to readdress the theoretical model. PMID- 11870774 TI - Generation of minimal protein identifiers of proteins from two-dimensional gels and recombinant proteins. AB - We describe the technical feasibility and methodology to characterize a protein by a minimal set of structural information generated by matrix assisted laser desorption/ionization (MALDI)-mass spectrometry, termed a "minimal protein Identifier" (MPI). MPIs can be determined for proteins from two-dimensional gels and recombinant proteins and can be used to compare and identify proteins from these sources. PMID- 11870775 TI - Proteomic analysis of the endoplasmic reticulum from developing and germinating seed of castor (Ricinus communis). AB - Endoplasmic reticulum (ER) has been prepared and analysed from germinating and developing castor bean endosperm. A combination of one- and two-dimensional (1-D and 2-D) gel electrophoresis was used to study the complexity of sample and protein differences between the two stages. The ER of the developing oilseed is central to the synthesis, sorting and storage of protein and lipid reserves while the germinating seed is concerned with their degradation. Sample complexity has been reduced by separation of ER proteins into lumenal, peripheral membrane and integral membrane subfractions. Membrane proteins pose specific problems in aggregation and binding to passive surfaces. We have overcome this by collection of membranes at density gradient interfaces and by silanization of plastic ware. Several major components have been identified from 1-D gels by N-terminal sequencing and matrix-assisted laser desorption/ionization (MALDI) peptide mass fingerprints. These include protein disulphide isomerase (PDI), calreticulin and developing-ER-specific oleate-12-hydroxylase involved in the biosynthesis of ricinoleic acid. In excess of 300 spots are detectable in each developmental fraction by high sensitivity 2-D gels. This is the first 2-D electrophoretic analysis of plant ER. These gels reveal significant differences between germinating and developing ER. Preparative loading 2-D gels of germinating ER have been run and 14 selected spots characterized by quadrupole time of flight tandem mass spectrometry (Q-TOF MS/MS). Ten of these proteins were assigned function on the basis of identity with existing castor database entries, or by homology with other species. Two proteins, aspartate proteinase precursor and N carbamyl-L-aminohydrolase-like protein, appear to be absent from developing profiles. Most of the proteins identified are concerned with roles in protein processing and storage, and lipid metabolism which occur in the ER. Data from three of the assigned spots included unidentified peptides indicating the presence of more than one protein in these spots following 2-D electrophoresis. More extensive analysis will have to await developments in genomics but the basic separation technologies to simplify sample identity for a plant ER preparation have been established. PMID- 11870776 TI - Biochemical dissection of the mitochondrial proteome from Arabidopsis thaliana by three-dimensional gel electrophoresis. AB - We report a subdivision of the mitochondrial proteome into defined sets of proteins, which is based on the combination of three different gel electrophoresis procedures. First, Blue-native polyacrylamide gel electrophoresis is employed to separate mitochondrial protein complexes. The protein complexes are electroeluted and completely detached from Coomasssie blue. Subsequently the subunits of the protein complexes are separated by isoelectric focusing and finally by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis. The resolution capacity of the procedure is demonstrated for the ATP synthase complex, the cytochrome c reductase complex and the preprotein translocase of the outer mitochondrial membrane (the TOM complex). The method allows the separation of isoforms of subunits forming part of protein complexes, whose occurrence seems to be rather a rule than an exception in higher eukaryotes. Furthermore, extremely hydrophobic proteins are detectable on the gels. PMID- 11870777 TI - Efficient peptide mapping and its application to identify embryo proteins in rice proteome analysis. AB - Using direct N-terminal analysis, only 31 N-terminally unblocked proteins out of 100 rice embryo proteins could be identified. To obtain protein sequence information for the remaining 69 blocked proteins, we developed a simple, efficient and rapid method. Using this method, we determined the peptide maps of 20 proteins per day in 10 pmol amounts. Applying this method to rice proteome analysis, we determined the internal sequences of all 69 blocked proteins. A total of 28 proteins out of 100 analyzed showed sequence similarity to the proteins with known functions in the SWISS-PROT and NCBI databases. Alternatively, we also used peptide mass fingerprinting determined by matrix assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF MS) to identify the rice proteins separated by two-dimensional electrophoresis (2 DE). Although peptide-mass fingerprinting is a high-throughput method, we could not easily identify all the rice proteins or genes by this method, because the complete database information on rice, is not yet available and many proteins are post-translationally modified. Therefore, at present, the improved peptide mapping method as we report here is considered to be very useful in rice proteome analysis, especially for blocked proteins. PMID- 11870778 TI - Iron-regulated proteins in Phanerochaete chrysosporium and Lentinula edodes: differential analysis by sodium dodecyl sulfate polyacrylamide gel electrophoresis and two-dimensional polyacrylamide gel electrophoresis profiles. AB - Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and two dimensional gel electrophoresis (2-DE) were used to identify iron-responsive proteins in the white-rot species (Phanerochaete chrysosporium and Lentinula edodes), by comparing the differential patterns of cellular and membrane proteins obtained from iron-sufficient and iron-deficient mycelia. Six cellular proteins induced by iron restriction have been observed in SDS-PAGE for P. chrysosporium and twelve for L. edodes. In 2-DE, the numbers of iron-restricted induced proteins were 12 and 9, respectively, in a resolution range of 15-60 kDa and pI 4.5-8.1. SDS-PAGE for the plasma membrane protein did not show differences, whereas the outer-membrane protein profile showed 6 and 5 proteins induced by iron depletion in P. chrysosporium and L. edodes, respectively. The results presented here are important data to unravel mechanisms of biosynthesis and/or transport of the iron-complexing agents in ligninolytic fungi and to further correlate them to the ligninolytic processes. PMID- 11870779 TI - Heat stress-induced loss of eukaryotic initiation factor 5A (eIF-5A) in a human pancreatic cancer cell line, MIA PaCa-2, analyzed by two-dimensional gel electrophoresis. AB - Alterations of intracellular proteins during the process of heat stress-induced cell death of a human pancreatic cancer cell line, MIA PaCa-2, were investigated using two-dimensional gel electrophoresis (2-DE), agarose gel electrophoresis, and cell biology techniques. Incubation of MIA PaCa-2 at 45 degrees C for 30 min decreased the cell growth rate and cell viability without causing chromosomal DNA fragmentation. Incubation at 51 degrees C for 30 min suppressed cell growth and again led to death without DNA fragmentation. The cell death was associated with the loss of an intracellular protein of M(r) 17,500 and pI 5.2 on 2-DE gel. This protein was determined to be eukaryotic initiation factor SA (eIF-5A) by microsequencing of the N-terminal region of peptide fragments obtained by cyanogen bromide treatment of the protein blotted onto a polyvinylidene difluoride (PVDF) membrane. The sequences detected were QXSALRKNGFVVLKGRP and STSKTGXHGHAKVHLVGID, which were homologous with the sequence of eIF-5A from Gln 20 to Pro 36 and from Ser 43 to Asp 61, respectively. Furthermore, the result of sequencing suggested that the protein was an active form of hypusinated eIF-5A, because Lys 46 could be detected but not Lys 49, which is the site for hypusination. These results suggest that loss of the active form of eIF-5A is an important factor in the irreversible process of heat stress-induced death of MIA PaCa-2 cells. PMID- 11870780 TI - Inhibition of heat-induced phosphorylation of stathmin by the bioflavonoid quercetin. AB - Effects of quercetin on heat-induced phosphorylation of stathmin in JURKAT cells were examined. Two-dimensional electrophoresis of stathmin showed that heat shock increases mono- and diphosphorylation of stathmin. Monophosphorylation induced by heat shock was inhibited by the presence of 0.1 mM quercetin, but not by the presence of 0.1 microM staurosporine. Immunoblot analysis of phosphorylated stathmin showed that heat-induced phosphorylation at Ser-38 was inhibited by quercetin but not by staurosporine. Quercetin enhanced heat-induced tyrosine phosphorylation of MAP kinase. These observations indicate that quercetin inhibits heat-induced phosphorylation at Ser-38 of stathmin but mitogen-activated protein (MAP) kinase is not involved in its phosphorylation. PMID- 11870784 TI - Non-ablative laser treatment of facial rhytides: a comparison of 1450-nm diode laser treatment with dynamic cooling as opposed to treatment with dynamic cooling alone. AB - BACKGROUND AND OBJECTIVE: Non-ablative dermal remodeling has been shown to create new dermal collagen. This is thought to occur secondary to a laser-induced injury to the skin. Other mechanisms of injury may lead to similar results. The aim of this study was to evaluate the efficacy and complication rate of a 1450-nm diode laser and compare clinical effect when the laser is used in conjunction with cryogen cooling as compared to the use of cryogen cooling alone. STUDY DESIGN/MATERIALS AND METHODS: Twenty subjects, skin types I-IV, age range 42-70 years, with Class I and II rhytides were enrolled in the study. Subjects were treated with 2-4 laser treatments and cryogen cooling on one side of their face, while the contralateral side was treated with cryogen cooling alone. Subjects were evaluated six months after their final treatment. RESULTS: Thirteen subjects showed clinical improvement on the laser/cryogen treated side. No subjects were noted to have any improvement at the cryogen alone side. CONCLUSION: The 1450-nm diode laser can lead to non-ablative improvement of rhytides. This effect appears to be a direct laser induced effect. PMID- 11870785 TI - Effects of intense pulsed light on sun-damaged human skin, routine, and ultrastructural analysis. AB - BACKGROUND AND OBJECTIVE: New, non-ablative methods can be used in skin rejuvenation. Histologic analysis of non-ablative IPL effects on facial, sun damaged skin. STUDY DESIGN/MATERIALS AND METHODS: Five female subjects, wrinkle class I or II and Fitzpatrick skin types I, II, and III. IPL treatment: once monthly, 560-nm cut-off filters, spot size 8x35 mm, 28-36 J/cm. Routine histology or electron microscopy on 2-mm punches, before treatment and then 1 week, 3 months, and 12 months. RESULTS: Pre-treatment specimens contained solar elastosis and perifollicular lymphoid infiltrates. Collagen and elastic fibers appeared unaffected by treatment. At 1-week, Demodex organisms appeared coagulated. CONCLUSIONS: Under these conditions, IPL induces minimal morphologic changes in mildly sun-damaged skin. Some esthetic improvement may be secondary to clearing of Demodex organisms and reduction of associated lymphocytic infiltrate. PMID- 11870786 TI - Facial resurfacing in patients with Fitzpatrick skin type IV. AB - BACKGROUND AND OBJECTIVES: Though post-inflammatory hyperpigmentation (PIH) is probably the most common complication of laser resurfacing and appears to correlate directly with the intensity of the patient's natural pigmentation, there is very little data that specifically addresses the risks of dyspigmentation in more darkly pigmented patients (Fitzpatrick skin types IV and above). The objective of this study was to evaluate the long-term dyspigmentation of patients with skin type IV having radial laser resurfacing. STUDY DESIGN/MATERIALS AND METHODS: A retrospective review of the clinical efficacy, incidence of dyspigmentation and other adverse effects, as well as the pre/post operative protocol of 22 patients with Fitzpatrick skin type IV who were a minimum of 1 year post-operative following facial laser resurfacing. RESULTS: The average patient achieved greater than 50% improvement, indicating adequate treatment being delivered. PIH occurred in 68% of patients, starting 1 month post operative and lasting 3.8 months. There was no correlation to pre-treatment or type of laser used as far as incidence of PIH. True hypopigmentation was not seen in this group of 22 patients. CONCLUSIONS: PIH is the most common complication of facial resurfacing in patients with skin type IV. It is not preventable by choice of laser or skin care regimen pre-operative, but appears to respond to appropriate treatment once it has developed. PMID- 11870787 TI - Use of a novel erbium laser in a Yucatan minipig: a study of residual thermal damage, ablation, and wound healing as a function of pulse duration. AB - BACKGROUND AND OBJECTIVE: Theoretical models show that varying pulse duration influences residual thermal damage in erbium YAG skin resurfacing. Accordingly, our objective was to compare residual thermal damage, ablation, tissue shrinkage, and wound healing between a variable pulsewidth erbium YAG laser and a popular CO2 resurfacing laser. STUDY DESIGN/MATERIALS AND METHODS: The erbium laser delivered a typical ablative pulse (250 microseconds), followed by a heating pulse of variable duration. Pulse durations for specific coagulation depths were selected based on existing heat transfer models. The bilateral flanks of one Yucatan pig were irradiated. Eight sites were treated per group. Biopsies were performed just after treatment and 1, 3, 7, 21, and 60 days postoperatively. RESULTS: Just after irradiation, gross examination of "cold" (without a coagulation pulse) erbium sites showed a reddish papillary dermis consistent with conventional erbium laser ablation. Two and three pass CO2 sites showed uniform surface yellowing. The longer pulsewidth ("hot") erbium groups showed only slight surface yellowing. Biopsies showed immediate thermal damage that increased with erbium pulse duration; however, actual residual thermal damage (RTD) was sometimes less than that predicted by the laser control panel. All wounds healed uneventfully by 14 days. CONCLUSIONS: An erbium laser with a variable macropulse pulsewidth was capable of achieving RTD of up to 80 mum. Even greater RTD depths may be obtainable with future manipulations of fluence and pulse duration. PMID- 11870788 TI - Evaluating the efficacy of using a short-pulsed erbium:YAG laser for intraoperative resurfacing of surgical wounds. AB - BACKGROUND AND OBJECTIVE: There are many studies demonstrating the aesthetic benefits of resurfacing a wound 4-8 weeks following surgical closure. Several anecdotal reports have been published stating that resurfacing a wound at the time of closure is also of cosmetic benefit. Our study was designed to evaluate the effects of resurfacing wound edges at the time of surgical closure. STUDY DESIGN/MATERIALS AND METHODS: Ten patients with skin types I-III undergoing reconstruction on the head or neck using a complex linear closure of at least 4 cm in length were enrolled in the study. Following the placement of subcutaneous sutures and an intradermal running suture, a split scar study was designed. Half of the wound was left as a control and half of the wound was resurfaced with a short pulsed Erbium:YAG laser. Both sides were treated with a hydrogel dressing. Follow-up evaluations were performed 1 week, 1 month, and 3 months post operatively. The wound was evaluated for any adverse reactions. The degree of erythema, textural misalignment or tissue mismatch, and overall aesthetic appearance of both sides of the scar were evaluated and scored. RESULTS: There were no incidents of infection, dehiscence, hematoma, necrosis, or reaction to the dressing during the study. There was a trend towards greater erythema in the resurfaced half of the scar at the 1 week and 1 month evaluation. There was a trend towards less tissue mismatch and better overall aesthetic appearance of the side of the scar that had been resurfaced. The differences however were not statistically significant. CONCLUSION: Given the time and resources necessary to perform an intraoperative resurfacing procedure on the wound edges, it may be more reasonable to withhold resurfacing procedures for those few cases that may require it postoperatively (patients with a history of poor healing, highly sebaceous areas, etc.). When good operative technique is used, most surgical wounds on the head and neck heal very well with excellent aesthetic outcomes without any additional intervention. PMID- 11870790 TI - Free-electron laser and heat-conducting templates: a study of reducing cutaneous lateral thermal damage. AB - BACKGROUND AND OBJECTIVES: We developed novel heat-conducting templates, and tested whether they could effectively remove damaging heat from the tissue during laser ablation. The reduction of lateral thermal damage during cutaneous incisional laser procedures should decrease the time in wound healing. In addition, we selected various infrared wavelengths to determine whether the template effects would be influenced by the laser penetration depth and the particular chromophore absorbing the laser light. STUDY DESIGN/MATERIALS AND METHODS: This study utilized the Free-Electron Laser at wavelengths of 3.0, 5.5, 6.45, 7.5, and 7.7 microm to produce 1.0 cm incisions on in vitro lightly pigmented human skin. At each of these wavelengths, copper, aluminum, glass, and Plexiglas heat conducting templates were tested. At wavelength 5.5 microm, the study was duplicated using in vitro darkly pigmented skin. Histological samples were evaluated using computerized morphometric analysis. RESULTS: The adjunct use of both the copper and aluminum templates provided a decrease in thermal damage at each wavelength. Using the copper template reduced lateral thermal damage an average of 67% with no apparent wavelength dependence. The aluminum template reduced thermal damage an average of 54% with no apparent wavelength dependence. The glass and Plexiglas templates did not reduce the lateral thermal damage. At 5.5 microm, no statistically significant difference in lateral thermal damage was observed between darkly and lightly pigmented tissues. CONCLUSIONS: Heat conducting templates are an effective new method to reduce lateral thermal damage from thermal laser incisions. PMID- 11870789 TI - MMPs and TIMP-1 are differentially expressed between acute murine excisional and laser wounds. AB - BACKGROUND AND OBJECTIVES: Matrix metalloproteinases (MMPs) and their physiological inhibitors, the tissue inhibitors of metalloproteinases (TIMPs) play significant roles in wound repair. The aim of this study was to determine whether MMP and TIMP-1 mRNA is differentially expressed between laser-created and excisional skin wounds. STUDY DESIGN/MATERIALS AND METHODS: Full-thickness wounds were created on the dorsal surface of Balb/c mice by either excision of the skin or with a CO2 laser. Animals were sacrificed at days 1, 3, 5, 7, 10, and 13, and their wounds processed for RNA isolation or histology and immunohistochemistry. RESULTS: Northern hybridization for MMP-3, -10, -13, and TIMP-1 mRNA demonstrated higher expression on days 1, 3, and 5 in the laser wounds as compared to the excisional wounds. Histological evaluation revealed a two day delay in the resurfacing of laser wounds as compared to excisional wounds. Immunohistochemistry of Day 5 and 7 excisional and laser wounds showed MMP-3 within the epidermal compartment and the neo-dermis. No remarkable differences were noted in the spatial distribution patterns of MMP-3 between wound types. CONCLUSIONS: Our findings demonstrate that MMPs and TIMP-1 mRNA expression is higher on days 1-5 post injury in full thickness laser-created wounds than in comparable same day excisional wounds. PMID- 11870791 TI - Zoon's balanitis treated with Erbium:YAG laser ablation. AB - BACKGROUND AND OBJECTIVE: Zoon's balanitis (ZB) is an uncommon, benign, idiopathic inflammatory condition affecting uncircumcised males. Definitive treatments include circumcision and carbon dioxide (CO2) laser ablation. We describe an alternative laser modality showing efficacy in the treatment of ZB. STUDY DESIGN/MATERIALS AND METHODS: We report the first case of ZB effectively treated with Erbium:YAG (Er:YAG) laser ablation. We review the pathogenesis and medical, surgical, and laser management of ZB. RESULTS: Our patient demonstrated a long-term clinical and histologic cure after Er:YAG laser ablation of ZB. CONCLUSIONS: As an alternative to circumcision or continuous wave CO2 laser, we propose a trial of Er:YAG laser ablation as a low risk, first line treatment of ZB. PMID- 11870792 TI - Hair removal with the long pulsed Nd:YAG laser: a prospective study with one year follow-up. AB - BACKGROUND AND OBJECTIVE: The aim was to investigate the efficacy, side effects, and the long-term results of a long pulsed Nd:YAG-Laser for hair removal in different hair colors and skin types. STUDY DESIGN/MATERIALS AND METHODS: We performed a prospective clinical study with 29 volunteers. Treatment was performed on the lower leg with a long pulsed Nd:YAG-Laser. Five test areas were treated 1-5 times in monthly intervals; one served as control. Follow-up investigations were performed at each session, and 3, 6, and 12 months after the last therapy. No depilatory treatment except shaving was allowed during the time of follow-up. Percentual hair loss, short- and long-term side effects, and pain during the treatment were evaluated. RESULTS: After one month, a hair loss of greater than 50% was found in 44.9% of the areas treated once. With up to five treatments, this percentage increased up to 71.5%. One year after therapy, a greater than 50% hair reduction was still present in 40% of the five-treatment areas and in 0% of the areas treated only once. There were no permanent side effects despite one small scar after a folliculitis. CONCLUSIONS: The long pulsed Nd:YAG is suitable to remove hair for more than 12 months effectively, although 4 5 sessions are necessary for these results. Blond hair can also be removed, although much less effective. No lasting side effects could be seen. Darker skin types or tanned skin can also be treated without side effects. A cooling may be advisable due to the pain reported by the volunteers. PMID- 11870793 TI - Pulsed dye laser and intralesional bleomycin for treatment of resistant viol hand warts. AB - BACKGROUND AND OBJECTIVE: Viral warts affect 7-10% of the population and are a major burden on time and resources of dermatology departments everywhere. Some warts prove resistant to multiple therapies, and this is particularly common in the immunosuppressed patient. Numerous treatments are available, but no one treatment has emerged as the treatment of choice. Bleomycin has been shown to be effective in treating warts, but administration can be difficult with risk of local complications. To demonstrate a new technique for easily and safely administrating bleomycin into warts, we undertook an open study to assess the practicality, efficacy, and tolerability of pulsed dye laser immediately followed by intralesional bleomycin in resistant viral hand warts. STUDY DESIGN/MATERIALS AND METHODS: Ten subjects, all with resistant viral hand warts of at least 3 years duration were recruited. Four subjects were on long-term immunosuppressant drugs. Eighteen warts treated in total. Area of the wart was anaesthetized with 1% lignocaine, then treated with a pulsed dye laser (7 mm spot, fluence 10 J/cm2). Immediately following this bleomycin (0.5 IU/ml) was injected into the base of the wart. Monthly follow-up and treatment until satisfactory clearance. RESULTS: Sixteen out of eighteen warts cleared (89%). Eight out of ten warts in the immunosuppressed subjects cleared. The remaining two warts responded partially to two treatments, but the patient was happy with the result and did not wish further treatment. All warts in the immunocompetent subjects cleared. No serious side effects were seen. CONCLUSIONS: The initial treatment of the wart with pulsed dye laser serves to "prepare" the wart for the bleomycin injection, which can then be given very easily. This ensures the drug is deposited into the base of the wart where it is most needed and minimises the risk of infiltration of normal skin or the operative environment. This method of combining pulsed dye laser and intralesional bleomycin appears to be a safe, rapid, well tolerated, and successful treatment for resistant hand warts. It has proven to be effective in warts in immunosuppressed patients and at difficult sites (e.g., subungual warts). These findings need confirmation from a larger controlled study. PMID- 11870794 TI - Clinical testing of a photoacoustic probe for port wine stain depth determination. AB - BACKGROUND AND OBJECTIVE: Successful laser treatment of port wine stain (PWS) birthmarks requires knowledge of lesion geometry. Laser parameters, such as pulse duration, wavelength, and radiant exposure, and other treatment parameters, such as cryogen spurt duration, need to be optimized according to epidermal melanin content and lesion depth. We designed, constructed, and clinically tested a photoacoustic probe for PWS depth determination. STUDY DESIGN/MATERIALS AND METHODS: Energy from a frequency-doubled, Nd:YAG laser (lambda=532 nm, tau(p)=4 nanoseconds) was coupled into two 1,500 mum optical fibers fitted into an acrylic handpiece containing a piezoelectric acoustic detector. Laser light induced photoacoustic waves in tissue phantoms and a patient's PWS. The photoacoustic propagation time was used to calculate the depth of the embedded absorbers and PWS lesion. RESULTS: Calculated chromophore depths in tissue phantoms were within 10% of the actual depths of the phantoms. PWS depths were calculated as the sum of the epidermal thickness, determined by optical coherence tomography (OCT), and the epidermal-to-PWS thickness, determined photoacoustically. PWS depths were all in the range of 310-570 microm. The experimentally determined PWS depths were within 20% of those measured by optical Doppler tomography (ODT). CONCLUSIONS: PWS lesion depth can be determined by a photoacoustic method that utilizes acoustic propagation time. PMID- 11870795 TI - Cooling gel improves pulsed KTP laser treatment of facial telangiectasia. AB - BACKGROUND AND OBJECTIVE: Pulsed KTP lasers effectively treat facial telangiectasia without purpura production. Transient side effects following treatment include erythema, edema, and vesiculation leading to crust formation. The aim of this study was to investigate the utility of an aqueous gel in reducing side effects associated with pulsed KTP laser treatment of facial telangiectasia. STUDY DESIGN/MATERIALS AND METHODS: Nineteen patients with extensive facial telangiectasias were treated with a pulsed KTP laser (Versapulse, Coherent, Palo Alto, CA). The laser was used with the water cooled handpiece chilled to 4C, a 4 mm spot size, a 10-millisecond pulse duration and a fluence of 9.5 J/cm2. One side of the face was treated with the laser using the cooling handpiece alone. The other side was treated using the cooling handpiece applied to a 2-mm film of aqueous gel spread over the treatment area. RESULTS: Treatment side effects, including pain, erythema, edema, vesiculation, and crusting were scored following treatment with and without the aqueous gel. Use of the aqueous gel in conjunction with the cooling handpiece decreased the incidence and severity of pain, erythema, edema, and crusting following pulsed KTP laser treatment of facial telangiectasia. Most patients demonstrated 50-75% clearance of their telangiectasias 1 month after one treatment session, and use of the gel did not alter the treatment efficacy. CONCLUSIONS: The application of an aqueous gel during pulsed KTP laser treatment of facial telangiectasia improves treatment associated side effects without affecting vessel clearance. PMID- 11870796 TI - Comparative pilot study evaluating the treatment of leg veins with a long pulse ND:YAG laser and sclerotherapy. AB - BACKGROUND AND OBJECTIVE: To date there have been very few side by side comparison studies of laser versus sclerotherapy in treating small leg veins. This study compares a long pulsed Nd:YAG laser with contact cooling to sclerotherapy for treating small diameter leg veins by evaluating objective and subjective clinical effects. STUDY DESIGN/MATERIALS AND METHODS: Twenty patients were selected with leg veins ranging from 0.25 to 3 mm at two comparable sites. One site was treated with long pulsed Nd:YAG laser and the other received sotradecol sclerotherapy. The patients followed up at 8 weeks for another possible laser retreatment and 3 months following the last treatment. Photographs were taken pre- and post-operatively and at each follow-up visit and used for objective comparative analysis. The patients also completed a Quality of Life survey. RESULTS: Improvement was tabulated from the photographic assessment on an improvement scale from 0 (no change)-4 (greater than 75% clearing). The laser treated areas averaged 2.50 and sclerotherapy treated sites averaged 2.30. Patient surveys show 35% preferred laser and 45% choose sclerotherapy. CONCLUSION: This pilot study demonstrates that the Lyra Long Pulse Nd:YAG laser can yield results similar to sclerotherapy in the treatment of small leg veins. PMID- 11870797 TI - Vascular response to laser photothermolysis as a function of pulse duration, vessel type, and diameter: implications for port wine stain laser therapy. AB - BACKGROUND AND OBJECTIVE: Treatment of port wine stains (PWS) by photothermolysis can be improved by optimizing laser parameters on an individual patient basis. We have studied the critical role of pulse duration (t(p)) on the treatment efficacy. STUDY DESIGN/MATERIALS AND METHODS: The V-beam laser (Candela) allowed changing t(p) over user-specified discrete values between 1.5 and 40 milliseconds by delivering a series of 100 microsecond spikes. For the 1.5 and 3 millisecond pulses, three spikes were observed at intervals t(p)/2 and for t(p)> or =6 milliseconds, four spikes separated by t(p)/3. The ScleroPlus laser (Candela) has a smooth output over its fixed 1.5 milliseconds duration. Blood vessels in the chick chorioallantoic membrane (CAM) were irradiated at fixed wavelength (595 nm), spot size (7 mm), radiant exposure (15 Jcm(-2)), and at variable t(p). The CAM contains an extensive microvascular network ranging from capillaries with diameter D<30 microm to blood vessels of D approximately 120 microm. The CAM assay allows real-time video documentation, and observation of blood flow in pre capillary arterioles (A) and post-capillary venules (V). Vessel injury was graded from recorded videotapes. Mathematical modeling was developed to interpret results of vessel injury when varying t(p) and D. A modified thermal relaxation time was introduced to calculate vessel wall temperature following laser exposure. RESULTS: Arterioles. For increasing t(p), overall damage was found to decrease. For fixed t(p), damage decreased with vessel size. Venules. For all D, damage was smaller than for corresponding arterioles. There was no dependence of damage on t(p). For given t(p), no variation of damage with D was observed. Photothermolysis due to spiked (V-beam) vs. smooth (Scleroplus) delivery of laser energy at fixed t(p) (1.5 milliseconds), showed similar vessel injuries for al values of D (P>0.05). CONCLUSIONS: The difference between initial arteriole and venule damage could be explained by the threefold higher absorption coefficient at 595 nm in (oxygen-poor!) arterioles. In human patients, PWS consist of ectatic venules (characterized by higher absorption), so that these considerations favor the use of 595-nm irradiation for laser photothermolysis. For optimal treatment of PWS it is proposed that t(p) be between 0.1 and 1.5 milliseconds. This is based on a modified relaxation time tau'(d), defined as the time required for heat conduction into the full thickness of the vessel wall, which is assumed to have a thickness DeltaD approximately 0.1D. The corresponding tau'(d) will be a factor of about six smaller than given in the literature. For vessels with D between 30 and 300 mum, tau'(d) ranges from 0.1 to 1.5 milliseconds. PMID- 11870798 TI - Different profiles of neuroendocrine cell differentiation evolve in the PC-310 human prostate cancer model during long-term androgen deprivation. AB - BACKGROUND: Neuroendocrine (NE) cells are androgen-independent cells and secrete growth-modulating peptide hormones via a regulated secretory pathway (RSP). We studied NE differentiation after long-term androgen withdrawal in the androgen dependent human prostate cancer xenograft PC-310. METHODS: Tumor-bearing nude mice were killed at 0, 2, 5, 7, 14, 21, 47, 84, and 154 days after castration. The half-life of the PC-310 tumor was 10 days, with a stable residual tumor volume of 30--40% after 21 days and longer periods of androgen deprivation. RESULTS: Proliferative activity and prostate-specific antigen serum levels decreased to zero after castration, whereas cell-cycle arrest was manifested by increased p27(kip1) expression. A temporary downregulation of androgen receptor (AR) expression was noted after androgen deprivation. The expression of chromogranin A, secretogranin III, and secretogranin V (7B2) increased 5 days after castration and later. Subsequently, pro-hormone convertase 1 and peptidyl alpha--amidating monooxygenase as well as vascular endothelial growth factor were expressed from 7 days after castration on. Finally, such growth factors as gastrin-releasing peptide and serotonin were expressed in a small part of the NE cells 21 days after castration, but strong expression was induced late during androgen deprivation, that is, 84 and 154 days after castration, respectively. CONCLUSIONS: Androgen deprivation of the NE-differentiated PC-310 model induced the formation of NE-differentiated AR(minus sign) and non-NE AR(+) tumor residues. The NE-differentiated cells actively produced growth factors via an RSP that may lead to hormone-refractory disease. The dormant non-NE AR(+) tumor cells were shown to remain androgen sensitive even after long-term androgen deprivation. In the PC-310 xenograft, time-dependent NE differentiation and subsequent maturation were induced after androgen depletion. The androgen dependent PC-310 xenograft model constitutes an excellent model for studying the role of NE cells in the progression of clinical prostate cancer. PMID- 11870799 TI - Role of family history and ethnicity on the mode and age of prostate cancer presentation. AB - BACKGROUND: This study was conducted to investigate the role of family history of prostate cancer and ethnic variation on age at diagnosis and the mode of prostate cancer presentation among a multiracial cohort of men in the Bronx and on Long Island. METHODS: A retrospective hospital-based study of 953 men (62% white, 24% African-American, 9% Hispanic, and 5% other ethnicities) with biopsy-confirmed prostate cancer diagnosed between 1991 and 1996. Data were collected between 1996 and 1998 using self-administered questionnaires that assessed age at diagnosis, ethnicity, family history of prostate cancer, and first indication of potential prostate cancer. RESULTS: Men with a family history of prostate cancer were diagnosed at an earlier mean age than those who lacked a family history (P<0.001). Prostate cancer patients with an affected father had a significantly lower mean age at diagnosis than those patients who indicated that at least one brother (but not their father) was affected (P<0.001). African-Americans reported a family history of prostate cancer more often than whites (P<0.01) and were younger at diagnosis (P<0.0001). Hispanic patients were less likely to be identified by screening and more likely to present with symptoms compared with whites (P<0.0001) and African-Americans (P<0.001). CONCLUSIONS: Men with an affected father were more likely to have disease diagnosed at an early age. The lower rates of presentation by prostate cancer screening of Hispanic men with prostate cancer suggests that increased surveillance may be warranted in this population. PMID- 11870800 TI - Establishment and characterization of androgen-independent human prostate cancer LNCaP cell model. AB - BACKGROUND: The acquisition of an androgen-independent phenotype is the most serious issue of prostate cancer treatment. Although several experimental cell models have been reported for studying androgen independence, they have limited applications related to hormone-refractory prostate cancer. To investigate the molecular mechanism of androgen-independent growth of prostate cancer, we established a useful LNCaP cell model that resembles the clinical scenario of hormone-refractory prostate cancer. METHODS: Androgen-sensitive LNCaP parental cells were continuously maintained in a regular cell-culture medium, that is, phenol red-positive RPMI 1640 medium supplemented with 5% fetal bovine serum and 1% glutamine. Upon passage, the androgen responsiveness of those cells decreased, to a level lower than that of parental cells. We examined the growth properties and androgen responsiveness of these different LNCaP cells in vitro and in vivo. Cytogenetic characteristics and expression of androgen receptors (ARs) and prostate-specific antigen (PSA) were determined. RESULTS: Upon continuous passage, the biological behavior of parental C-33 cells (passage number less than 33) was altered. C-81 cells (passage number higher than 81) clearly exhibited more aggressive growth and lower androgen responsiveness than C-33 and C-51 cells (passage number between 35 and 80) in vitro and in vivo. Nevertheless, all these cells expressed a similar level of functional AR protein as well as a similar genetic profile. Moreover, in a steroid-reduced culture condition, C-81 cells secreted a higher level of PSA than C-33 cells. CONCLUSIONS: Our LNCaP cell model closely recapitulates the progression of human prostate cancer from the androgen responsive to the hormone-refractory state under the androgen nondeprived condition. This cell model may provide the opportunity to understand the molecular mechanisms associated with the acquisition of androgen independence during human prostate cancer progression. PMID- 11870801 TI - Clinical stage T1c prostate cancer: pathologic outcomes following radical prostatectomy in black and white men. AB - BACKGROUND: The incidence of prostate cancer in black men is 50% to 70% higher than among age-matched white men. Black men have a twofold higher mortality rate and overall tend to have higher serum prostate-specific antigen (PSA) levels than white men. To determine whether racial differences exist in men whose prostate cancer was diagnosed based solely on an elevated serum PSA level, we compared clinical and pathologic features in black and white men undergoing radical prostatectomy (RP) for clinical stage T1c prostate cancer. METHODS: We used a prospectively collected database to identify all men undergoing RP for clinical T1c prostate cancer between July 1995 and October 2000. A total of 129 consecutive men (56 black men and 73 white men) were compared for age at diagnosis, serum PSA level, biopsy Gleason score, pathologic stage, RP specimen Gleason score, incidence of lymph node metastasis, and incidence of positive surgical margins. RESULTS: Statistically significant differences were not found by race in patients' ages, serum PSA levels, biopsy Gleason score, pathologic stage, incidence of lymph node metastases, or incidence of positive surgical margins. The RP specimen Gleason score was more heterogeneous in black men than white men (P=0.02). CONCLUSIONS: Racial differences in the incidence and mortality rate of prostate cancer are well known, but differences in the clinical and pathologic features between black and white men with prostate cancer identified solely based on an elevated serum PSA level with negative results on digital rectal examination (clinical stage T1c ) have been poorly studied. Our results suggest that men with clinical stage T1c prostate cancer have similar clinical and pathologic findings regardless of race. These results suggest that early-detection programs using serum PSA testing for prostate cancer in black men potentially can result in improvements in prostate cancer outcomes in this high risk group. PMID- 11870803 TI - Application of tissue-marking ink to prostate biopsy specimens. AB - BACKGROUND: When sextant prostate biopsy specimens are performed, noting the location from which cancer-containing cores were taken aids in treatment planning. However, many institutions include several cores in a single container, to cut costs. We have tried several methods of ink application and combining ink labeled biopsy specimens into fewer containers with the goal of maintaining information regarding location while minimizing the expense. METHODS: Several approaches to the application of tissue-marking ink to biopsy cores, core preparation methods, color combinations, and numbers of cores in each container were assessed. RESULTS: The ink adheres well to dry cores, but these cores often exhibit dehydration artifact. Placing the cores on a wet sponge avoids dehydration but causes ink spread. Excessive ink application occurs with eyedroppers and syringes but is avoided by touching each core with an ink moistened cotton swab. Application of 1% acetic acid to the inked core promotes congealing of the ink onto the tissue. Yellow, orange, and red ink are more difficult to distinguish than blue, black, or green. Deciphering distinct cores when three or more cores are combined is difficult, especially if cores are fragmented. CONCLUSIONS: With our current protocol, all biopsy specimens are placed on separate moistened gauze sponges. Specimens from the right side of the prostate are marked by green ink, and those from the left side are marked by black ink with a cotton swab. After applying 1% acetic acid to each core, the left and right cores from each location are placed in a single container. This method curbs pathology expenses and maintains tumor location information. PMID- 11870802 TI - Human prostatic carcinoma cells produce an increase in the synthesis of interleukin-6 by human osteoblasts. AB - BACKGROUND: The aim of this work was to evaluate the effect produced by conditioned medium from human prostatic carcinoma cell (PC-3) culture on human osteoblast (HOB) interleukin 6 (IL-6) synthesis. METHODS: PC-3 cells were cultured in Ham's F12K medium with 10% fetal calf serum (FCS) up to confluence. Medium was changed by Dulbecco modified Eagle medium (DMEM)/F12K (1:1) with 0.1% bovine serum albumin. Cells were cultured for 24 hr, and medium (PC-3-CM) was collected. HOBs were cultured up to confluence, and after 48 hr without FCS, medium was removed and PC-3-CM was added to the wells. After 24 hr, supernatant was collected for the determination of IL-6. In another experiment, HOBs were cultured up to confluence in Petri dishes, and after 48 hr without FCS, PC-3-CM or DMEM/F12K (1:1) was added. After different periods of time, medium was removed, and total RNA was extracted. IL-6 mRNA was quantified using reverse transcription polymerase chain reaction. RESULTS: PC-3-CM significantly enhanced IL-6 secretion into HOB culture supernatants (between 1,812% and 372%, depending on the osteoblastic line) with respect to HOBs cultured in DMEM/F12K. PC-3-CM also produced an increase in IL-6 mRNA levels in HOBs. CONCLUSIONS: Prostate carcinoma cells (PC-3) produce a factor or factors that enhance the synthesis and release of IL-6, a known activator of bone resorption. PMID- 11870804 TI - Androgen regulation of spermidine synthase expression in the rat prostate. AB - BACKGROUND: Spermidine synthase, an essential enzyme in the polyamine synthesis pathway, was identified as one of the androgen-response genes in the rat ventral prostate. Characterization of androgen regulation of spermidine synthase is important to the understanding of androgenic regulation of polyamine synthesis. METHODS: Full-length cDNA encoding rat spermidine synthase was isolated from a lambdaZAP cDNA phage library. Young male adult Sprague-Dawley rats were used for castration and androgen replacement. Northern blot and in situ hybridization were used to characterize gene expression. RESULTS: The amino acid sequence of rat spermidine synthase shares 99% and 94% identity with that of mouse and human spermidine synthase, respectively. Spermidine synthase gene is abundantly expressed and regulated by androgens in the ventral, dorsal, and lateral lobes of the rat prostate, and its expression is localized to the epithelial cells. Spermidine synthase also is regulated by androgens in the seminal vesicles but not in the muscle, brain, kidney, thymus, heart, or liver, suggesting that this enzyme is responsive to androgen in the male sex accessory organs only. The expression of spermidine synthase and two other enzymes involved in polyamine synthesis, S-adenosylmethionine decarboxylase and ornithine decarboxylase, are regulated by androgens coordinately. CONCLUSIONS: Spermidine synthase is most abundantly expressed and regulated by androgens in the prostatic epithelial cells, suggesting that regulation of spermidine synthase is likely a key step in coordinated androgen regulation of polyamine synthesis in the prostate. PMID- 11870805 TI - Effectiveness of mass screening for the prognosis of prostate cancer patients in Japanese communities. AB - BACKGROUND: A study was conducted to investigate the effectiveness of mass screening (MS) for prostate cancer in Japanese communities using the long-term follow-up results. METHODS: Subjects were 279 patients with prostate cancer detected by MS in Gunma prefecture (MS group) and 624 patients with prostate cancer who did not undergo screening (non-MS group). From the follow-up data, the prognoses were compared between the groups. RESULTS: The ratio of patients with Stage II cancer was higher for the MS group (59.1%) when compared to non-MS group (25.3%). As a whole, the prognosis of the MS group was more favorable than that of the non-MS group for relative survival rate. Prognosis was especially favorable for patients with Stage III prostate cancer in mass screening group considering the effect of lead-time bias. CONCLUSIONS: Early detection and longer survival of the patients with prostate cancer detected by mass screening suggested the effectiveness of prostate cancer screening. PMID- 11870806 TI - Individual bioequivalence testing under 2x3 designs. AB - In recent years, as more generic drug products become available, it is a concern not only whether generic drug products that have been approved based on the regulation of average bioequivalence will have the same quality, safety and efficacy as that of the brand-name drug product, but also whether the approved generic drug products can be used interchangeably. In its recent draft guidance, the U.S. Food and Drug Administration (FDA) recommends that individual bioequivalence (IBE) be assessed using the method proposed by Hyslop, Hsuan, and Holder to address drug switchability. The FDA suggests that a 2x4 cross-over design be considered for assessment of IBE, while a 2x3 cross-over design may be used as an alternative design to reduce the length and cost of the study. Little or no information regarding the statistical procedures under 2x3 cross-over designs is discussed in the guidance. In this paper, a detailed statistical procedure for assessment of IBE under 2x3 cross-over designs is derived. The main purpose of this paper, however, is to derive an IBE test under an alternative 2x3 design and show that the resulting IBE test is better than that under a 2x3 cross over design and is comparable to or even better than that under a 2x4 cross-over design. Our conclusions are supported by theoretical considerations and empirical results. Furthermore, a method of determining the sample sizes required for IBE tests to reach a given level of power is proposed. PMID- 11870807 TI - Estimating efficacy in clinical trials with clustered binary responses. AB - When non-compliance occurs in a clinical trial, it may be of interest to supplement the intent-to-treat analysis with an analysis of the efficacy (or biological effect) of therapy. Sommer and Zeger (1991) developed a method for estimating efficacy applicable to the case of a binary response variable and all or-none compliance that assumes independent subject responses. We extend this approach to accommodate within-cluster correlations as may be expected in a cluster-randomized design. The method is illustrated using data from a controlled village-randomized clinical trial conducted in Indonesia to investigate the effect of vitamin A supplementation on mortality in children. We find that within cluster correlations for these data are very small and that taking into account the clustering does not substantially affect inferences in this case. Additional calculations show that small within-cluster correlations (though larger than those found in the vitamin A data) may have a large impact on efficacy inferences. We also present the results of a simulation study that demonstrates the validness of the proposed approach for finite sample sizes. PMID- 11870808 TI - Assessment and monitoring in clinical trials when survival curves have distinct shapes: a Bayesian approach with Weibull modelling. AB - The comparison of Weibull distributions with unequal shape parameters, in the case of right censored survival data obtained from independent samples, is considered within the framework of Bayesian statistical methodology. The procedures are illustrated with the example of a mortality study where a new treatment is compared to a placebo. The posterior distributions about relevant parameters, which may provide support for a conclusion of clinical superiority of the treatment, and the predictive distributions, which may guide decision about early stopping at an interim analysis, are considered for a class of appropriate priors. PMID- 11870809 TI - Correcting for non-compliance of repeated binary outcomes in randomized clinical trials: randomized analysis approach. AB - We develop the randomized analysis for repeated binary outcomes with non compliance. A break randomization-based semi-parametric estimation procedure for both the causal risk difference and the causal risk ratio is proposed for repeated binary data. Although we assume the simple structural models for potential outcomes, we choose to avoid making any assumptions about comparability beyond those implied by randomization at time zero. The proposed methods can incorporate non-compliance information, while preserving the validity of the test of the null hypothesis, and even in the presence of non-random non-compliance can give the estimate of the causal effect that treatment would have if all individuals complied with their assigned treatment. The methods are applied to data from a randomized clinical trial for reduction of febrile neutropenia events among acute myeloid leukaemia patients, in which a prophylactic use of macrophage colony-stimulating factor (M-CSF) was compared to placebo during the courses of intensive chemotherapies. PMID- 11870810 TI - Analysis of the ratio of marginal probabilities in a matched-pair setting. AB - Statistical methods for testing and interval estimation of the ratio of marginal probabilities in the matched-pair setting are considered in this paper. We are especially interested in the situation where the null value is not one, as in one sided equivalence trials. We propose a Fieller-type statistic based on constrained maximum likelihood (CML) estimation of nuisance parameters. For a series of examples, the significance level of the CML test is satisfactorily close to the nominal level, while a Wald-type test is anticonservative for reasonable sample sizes. We present formulae for approximate power and sample size for the CML and Wald tests. The matched design is seen to have a clear advantage over the unmatched design in terms of asymptotic efficiency when the two responses of the pair are highly positively correlated. We recommend the CML method over the Wald method, especially for small or moderate sample sizes. PMID- 11870811 TI - A non-parametric method for the comparison of partial areas under ROC curves and its application to large health care data sets. AB - The receiver operating characteristic (ROC) curve is a statistical tool for evaluating the accuracy of diagnostic tests. Investigators often compare the validity of two tests based on the estimated areas under the respective ROC curves. However, the traditional way of comparing entire areas under two ROC curves is not sensitive when two ROC curves cross each other. Also, there are some cutpoints on the ROC curves that are not considered in practice because their corresponding sensitivities or specificities are unacceptable. For the purpose of comparing the partial area under the curve (AUC) within a specific range of specificity for two correlated ROC curves, a non-parametric method based on Mann-Whitney U-statistics has been developed. The estimation of AUC along with its estimated variance and covariance is simplified by a method of grouping the observations according to their cutpoint values. The method is used to evaluate alternative logistic regression models that predict whether a subject has incident breast cancer based on information in Medicare claims data. PMID- 11870812 TI - Estimation of the survival function for Gray's piecewise-constant time-varying coefficients model. AB - Gray's extension of Cox's proportional hazards (PH) model for right-censored survival data allows for a departure from the PH assumption via introduction of time-varying regression coefficients (TVC). For this model estimation of the conditional hazard rate relies on the inclusion of penalized splines. Cubic penalized splines tend to be unstable in the right tail of the distribution and thus quadratic, linear and piecewise-constant penalized splines may be a favourable choice. We derive a survival function estimator for one important member of the class of TVC models--a piecewise-constant time-varying coefficients (PC-TVC) model. Using the first-order Taylor series approximation we also derive an estimate for the variance of the log-transformed and log(-log)-transformed survival function, which in turn leads to estimated confidence limits on the corresponding scales of the survival function. Accuracy in estimating underlying survival times and survival quantiles is assessed for both Cox's and Gray's PC TVC model using a simulation study featuring scenarios violating the PH assumption. Finally, an example of the estimated survival functions and the corresponding confidence limits derived from Cox's PH and Gray's PC-TVC model, respectively, is presented for a liver transplant data set. PMID- 11870813 TI - Testing model fit in longitudinal data analysis against alternatives with omitted covariates. AB - Several types of common model misspecifications can be re-formulated as problems of omitted covariates. These include situations with unmeasured confounders, measurement errors in observed covariates and informative censoring. Longitudinal data present special opportunities for detecting omitted covariates that are related to the observed ones differently across time than across individuals. This situation arises with period and cohort effects, as well as with usual formulations of classical measurement error in observed covariates. In this article we focus on testing for the existence of omitted covariates in longitudinal data analysis when models are fit by generalized estimation equations. When omitted covariates are present, specification of the correct link function conditionally on only observed covariates under the alternative usually involves complicated numerical integration. We propose a quasi-score test statistic that avoids the need to fit such alternative models. The statistic is asymptotically chi-square distributed under the null hypothesis of no omitted covariates with degrees of freedom determined by the assumed alternative structure. We study the significance level and the power of the quasi-score test in linear and logistic regression models. The test is then applied to an analysis of excessive daytime sleepiness. PMID- 11870814 TI - Maximum likelihood estimation of a survival function with a change point for truncated and interval-censored data. AB - This paper considers estimation of a survival function when there exists a change point and the survival time of interest is defined as elapsed time between two related events. Furthermore, there exists censoring on observations on the occurrences of both events and truncation on observations on the occurrence of the second event and thus the survival time of interest. To obtain the maximum likelihood estimator of a survival function, an EM algorithm is developed when the survival function is completely unknown before the change point and known up to a vector of unknown parameters after the change point. The idea is a generalization of that discussed in Moeschberger and Klein. Simulations and an example are used to evaluate and illustrate the algorithm. PMID- 11870815 TI - Identification of significant host factors for HIV dynamics modelled by non linear mixed-effects models. AB - Non-linear mixed-effects models are powerful tools for modelling HIV viral dynamics. In AIDS clinical trials, the viral load measurements for each subject are often sparse. In such cases, linearization procedures are usually used for inferences. Under such linearization procedures, however, standard covariate selection methods based on the approximate likelihood, such as the likelihood ratio test, may not be reliable. In order to identify significant host factors for HIV dynamics, in this paper we consider two alternative approaches for covariate selection: one is based on individual non-linear least square estimates and the other is based on individual empirical Bayes estimates. Our simulation study shows that, if the within-individual data are sparse and the between individual variation is large, the two alternative covariate selection methods are more reliable than the likelihood ratio test, and the more powerful method based on individual empirical Bayes estimates is especially preferable. We also consider the missing data in covariates. The commonly used missing data methods may lead to misleading results. We recommend a multiple imputation method to handle missing covariates. A real data set from an AIDS clinical trial is analysed based on various covariate selection methods and missing data methods. PMID- 11870816 TI - Combining regression and association modelling for longitudinal data on bacterial carriage. AB - A longitudinal data set, from the Finnish Otitis Media (FinOM) Studies, reporting carriage or non-carriage of Streptococcus pneumoniae at 2, 3, 4, 5, 6, 9, 12, 15 and 18 months of age of 329 children living in Tampere, Finland, is analysed. A logistic regression model on five time varying explanatory variables is fitted. The temporal association between presence at different ages is measured by dependence ratios and the structure of these is shown to be well described by a model indicating that roughly 10 per cent of the children are not susceptible to the bacteria, while for those that are susceptible, carriage status at a future observation age is conditionally independent of past observed statuses, given the present status. The dependence ratios between carriage at adjacent observation ages decay exponentially with age. Maximum likelihood estimates are obtained for the parameters of the full model, which is the combination of the marginal logistic regression and the association models. The parameter estimates of the full model, strengthened by non-testable Markov assumptions, are used for assessing the median duration of carriage and the acquisition rate as functions of age. PMID- 11870817 TI - Comparing results of large clinical trials to those of meta-analyses. AB - We consider methods for assessing agreement or disagreement between the results of a meta-analysis of small studies addressing a clinical question and the result of a large clinical trial (LCT) addressing the same clinical question. We recommend basing conclusions about agreement upon the difference between the two results (relative risk, log-odds ratio or similar summary statistic), in the light of the estimated standard error of that difference. To estimate the standard error of the meta-analytic result we recommend a random effects analysis, and where a between-studies variance component is found, that component of variance should be used twice: once in the estimated standard error for the meta-analytic result and again in the standard error of the LCT result (augmenting the internal standard error of that statistic). Such broadening of the standard error reduces the appearance of disagreement. We also offer a critique of a different published approach, which is based on consistency of findings of statistical significance, a matter of how the two results regard zero, which is a poor measure of how closely they agree with each other. PMID- 11870818 TI - A seminar series in applied biostatistics for clinical research fellows, faculty and staff. AB - Familiarity with experimental design and statistical analysis techniques is necessary for investigators to conduct state-of-the-art research in clinical medicine. A wide range of options for biostatistical training is available. These include formal academic courses for credit, one-on-one education offered under the guise of consultation, self-paced textbook or computer learning, intensive short courses, portions of extended courses spanning an assortment of other subjects, and a continuing series of seminars or lectures covering selected topics in statistics. The latter structure is described in this article. We offered an eight-week lecture series focusing on a new topic in biostatistics each week. The course targeted clinical faculty, postdoctoral fellows, residents, and other interested research staff at a large medical research institution. The series focused on interpretation of medical journal articles with regard to statistical issues. Another objective was to provide information to increase a participant's expertise in the design of research studies. Guidance was offered for effective communication with collaborating biostatisticians. The format and contents of the seminar series are described. Aspects of the course that proved to be successful are discussed. PMID- 11870819 TI - Bayesian estimation of the proportion of treatment effect captured by a surrogate marker. AB - Surrogate endpoints in clinical trials are biological markers or events observable earlier than the clinical endpoints (such as death) that are actually of primary interest. The "proportion of treatment effect captured" by a surrogate endpoint (PTE) is a frequentist measure intended to address the question of whether trials based on a surrogate endpoint reach the same conclusions as would have been reached using the true endpoint. The question of inferential interest is whether PTE for a given marker exceeds some threshold value, say 0.5. Calculating PTE requires fitting two different models to the same data. We develop a Markov chain Monte Carlo based method for estimating the Bayesian posterior distribution of PTE. The new method conditions on the truth of a single model. Obtaining the full posterior distribution enables direct statements such as "the posterior probability that PTE >0.5 is 0.085". Furthermore, credible sets do not depend on asymptotic approximations and can be computed using data sets for which the frequentist methods may be inaccurate or even impossible to apply. We illustrate with Bayesian proportional hazards models for clinical trial data. As a by-product of developing the Bayesian method, we show that the frequentist estimate of PTE also may be computed from quantities in a single model and calculate frequentist confidence intervals for PTE that tend to be narrower than those produced by standard methods but that provide equally good coverage. PMID- 11870820 TI - Sample size calculations for comparative studies of medical tests for detecting presence of disease. AB - Technologic advances give rise to new tests for detecting disease in many fields, including cancer and sexually transmitted disease. Before a new disease screening test is approved for public use, its accuracy should be shown to be better than or at least not inferior to an existing test. Standards do not yet exist for designing and analysing studies to address this issue. Established principles for the design of therapeutic studies can be adapted for studies of screening tests. In particular, drawing upon methods for superiority and non-inferiority studies of therapeutic agents, we propose that confidence intervals for the relative accuracy of dichotomous tests drive the design of comparative studies of disease screening tests. We derive sample size formulae for a variety of designs, including studies where patients undergo several tests and studies where patients receive only one of the tests under evaluation. Both cohort and case-control study designs are considered. Modifications to the confidence intervals and sample size formulae are discussed to accommodate studies where, because of the invasive nature of definitive testing, true disease status can only be obtained for subjects who are positive on one or more of the screening tests. The methods proposed are applied to a study comparing a modified pap test to the conventional pap for cervical cancer screening. The impact of error in the gold standard reference test on the design and evaluation of comparative screening test studies is also discussed. PMID- 11870821 TI - Analytic methods for comparing two dichotomous screening or diagnostic tests applied to two populations of differing disease prevalence when individuals negative on both tests are unverified. AB - Two dichotomous screening tests may be compared by applying both tests to all members of a sampled population. For individuals with a positive result on either test the disease status may be verified by a reference standard, but for individuals negative on both tests the disease status may be unverified because the probability of disease is so low that further investigation is costly, unacceptable and perhaps unethical. If the tests have been applied to samples from two populations which have different disease prevalences then unbiased estimates of the true positive and false positive rates of each test, the prevalences in the two populations, and two parameters representing dependence between the two tests can be estimated using maximum likelihood methods. The methods are based on the assumption that the sensitivities and specificities of the two tests, and the dependencies between the tests, are independent of prevalence. A test of goodness of fit provides a test of this. PMID- 11870822 TI - Some further results on incorporating risk factor information in assessing the dependence between paired failure times arising from case-control family studies: an application to prostate cancer. AB - In a typical case-control family study, detailed risk factor information is often collected on cases and controls, but not on their relatives for reasons of cost and logistical difficulty in locating the relatives. The impact of missing risk factor information for relatives on estimation of the strength of dependence between the disease risk of pairs of relatives is largely unknown. In this paper, we extend our earlier work on estimating the dependence of ages at onset between paired relatives from case-control family data to include covariates on cases and controls, and possibly relatives. Using population-based case-control families as our basic data structure, we study the effect of missing covariates for relatives and/or cases and controls on the bias of certain dependence parameter estimators via a simulation study. Finally we illustrate various analyses using a case control family study of early onset prostate cancer. PMID- 11870823 TI - Statistical approaches to establishing vaccine safety. AB - In a vaccine safety trial, the primary interest is to demonstrate that the vaccine is sufficiently safe, rejecting the null hypothesis that the relative risk of an adverse event attributable to the new vaccine is above a prespecified value, greater than one. We evaluate the exact probability of type I error of the likelihood score test, with sample size determined by normal approximation, by enumeration of the binomial outcomes in the rejection region and show that it exceeds the nominal level. In the case of rare adverse events, we recommend the Poisson approximation as an alternative and develop the corresponding conditional and unconditional tests. We give sample size and power calculations for these tests. We also propose optimal randomization strategies which either (i) minimize the total number of adverse cases or (ii) minimize the expected number of subjects when the vaccine is unsafe. We illustrate the proposed methods using a hypothetical vaccine safety study. PMID- 11870824 TI - Semi-parametric models of long- and short-term survival: an application to the analysis of breast cancer survival in Utah by age and stage. AB - A flexible class of semi-parametric survival models is proposed that takes account of long- and short-term covariate effects in cancer survival. The diversity of responses described by the models include non-proportional and crossing survival curves as well as a fraction of long-term survivors. Restricted non-parametric maximum likelihood estimation procedures (RNPMLE) are developed to provide point estimates, confidence intervals and tests for the models. Numerical algorithms to fit semi-parametric survival models are emphasized. The methods are applied to analyse post-treatment survival of breast cancer patients diagnosed in Utah by age and stage. PMID- 11870825 TI - Regression-based variable clustering for data reduction. AB - In many studies it is of interest to cluster states, counties or other small regions in order to obtain improved estimates of disease rates or other summary measures, and a more parsimonious representation of the country as a whole. This may be the case if there are too many to summarize concisely, and/or many regions with a small number of cases. By merging the regions into larger geographic areas, we obtain more cases within each area (and hence lower standard errors for parameter estimates), as well as fewer areas to summarize in terms of disease rates. The resulting clusters should be such that regions within the same cluster are similar in terms of their disease rates. In this paper we present a clustering algorithm which uses data at the subject-specific level in order to cluster the original regions into a reduced set of larger areas. The proposed clustering algorithm expresses the clustering goals in terms of a regression framework. This formulation of the problem allows the regions to be clustered in terms of their association with the response, and confounding variables measured at the subject-specific level may be easily incorporated during the clustering process. Additionally, this framework allows estimation and testing of the association between the areas and the response. The statistical properties and performance of the algorithm were evaluated via simulation studies, and the results are promising. Additional simulations illustrate the importance of controlling for confounding variables during the clustering process, rather than after the clusters are determined. The algorithm is illustrated with data from the Cardiovascular Health Study. Although developed with a specific application in mind, the method is applicable to a wide range of problems. PMID- 11870827 TI - Abdominal imaging with a modular combination of spin and gradient echoes. AB - MR-CAT (combined acquisition technique), a modular hybrid imaging concept, was introduced recently. In this article it is demonstrated that the CAT principles can be applied to form a versatile combination of spin and gradient echoes for abdominal imaging. This CAT approach, which essentially integrates RARE and EPI modules in a sequential fashion, was used to implement a set of segmented and single-shot RARE/EPI-CAT imaging techniques. CAT was used in in vivo studies to perform high-resolution abdominal imaging in five healthy subjects. The results demonstrate the feasibility of abdominal imaging using the proposed CAT approach and the potential of this technique to reduce imaging time while preserving image quality. PMID- 11870826 TI - A quasi-exact method for the confidence intervals of the difference of two independent binomial proportions in small sample cases. AB - In this paper we propose a quasi-exact alternative to the exact unconditional method by Chan and Zhang (1999) estimating confidence intervals for the difference of two independent binomial proportions in small sample cases. The quasi-exact method is an approximation to a modified version of Chan and Zhang's method, where the two-sided p-value of an observation is defined by adding to the one-sided p-value the sum of all probabilities of more "extreme" events in the unobserved tail. We show that distinctively less conservative interval estimates can be derived following the modified definition of the two-sided p-value. The approximations applied in the quasi-exact method help to simplify the computations greatly, while the resulting infringements to the nominal level are low. Compared with other approximate methods, including the mid-p quasi-exact methods and the Miettinen and Nurminen (M&N) asymptotic method, our quasi-exact method demonstrates much better reliability in small sample cases. PMID- 11870829 TI - Reproducibility of 3D proton spectroscopy in the human brain. AB - The inter- and intrasubject reproducibility of the metabolite levels of N acetylaspartate (NAA), creatine (Cr), and choline (Cho), obtained with three dimensional (3D) multivoxel proton spectroscopy (1H-MRS), was analyzed in eight healthy volunteers. Serial, back-to-back measurements on a phantom showed the methodology and instrumentation to be highly reproducible, with a median coefficient of variation (CV) of 3.8%. In the human brain, the metabolite levels' variability was larger, with intrasubject median CVs for a total of 1876 signal voxels of 13.8%, 18.5%, and 20.1% for NAA, Cr, and Cho, respectively. These variations possibly arise from small, unavoidable, +/-1-2 mm volume-of-interest (VOI) repositioning uncertainties, which vary each 0.75-cm(3) voxel's partial fluid/gray/white-matter fractions. Comparing the CVs between eight subjects in a total of 324 selected voxels gave total interindividual CVs of 15.6%, 23.3%, and 24.4%, compared with intraindividual CVs in the same voxels of 14.4%, 14.8%, and 15.3%, for NAA, Cr, and Cho, respectively. Replacing the signal(s) from each voxel by the average of itself with its six canonical neighbors reduces the intrasubject median CVs to 8.3%, 9.5%, and 9.7%. The measurement uncertainties can be reduced at a cost of either spatial resolution (by using larger voxels) or time (by performing serial follow-ups). PMID- 11870828 TI - Combined MR proton lung perfusion/angiography and helium ventilation: potential for detecting pulmonary emboli and ventilation defects. AB - Three-dimensional (3D) perfusion imaging allows the assessment of pulmonary blood flow in parenchyma and main pulmonary arteries simultaneously. MRI using laser polarized (3)He gas clearly shows the ventilation distribution with high signal to-noise ratio (SNR). In this report, the feasibility of combined lung MR angiography, perfusion, and ventilation imaging is demonstrated in a porcine model. Ultrafast gradient-echo sequences have been used for 3D perfusion and angiographic imaging, in conjunction with the use of contrast agent injections. 2D multiple-section (3)He imaging was performed subsequently by inhalation of 450 ml of hyperpolarized (3)He gas. The MR techniques were examined in a series of porcine models with externally delivered pulmonary emboli and/or airway occlusions. With emboli, perfusion deficits without ventilation defects were observed; airway occlusion resulted in matched deficits in perfusion and ventilation. High-resolution MR angiography can unambiguously reveal the location and size of the blood emboli. The combination of the three imaging methods may provide complementary information on abnormal lung anatomy and function. PMID- 11870830 TI - In vivo GABA editing using a novel doubly selective multiple quantum filter. AB - A novel multiple quantum filtering method is proposed that uses a doubly selective pulse termed Delays Alternating with Nutations for Tailored Excitation (DANTE) for multiple quantum preparation. This method selectively prepares GABA-3 and GABA-4 into a multiple quantum state and suppresses all other resonances at 3.0 ppm in each single scan. Phantom tests demonstrated excellent GABA signal retention and complete suppression of overlapping metabolites. It is shown using numerical simulations that overlapping macromolecules are suppressed because the frequency of the first upfield 2pi rotation of the doubly selective DANTE pulse coincides with that of the macromolecules at 1.72 ppm. Excellent suppression of overlapping macromolecules was demonstrated in vivo. Using this method the concentration of GABA in the occipital lobe of healthy volunteers was measured to be 1.21 +/- 0.28 micromol/mL (mean +/-SD, N = 9). PMID- 11870831 TI - Biexponential diffusion attenuation in the rat spinal cord: computer simulations based on anatomic images of axonal architecture. AB - Water diffusion in neurological tissues is known to possess multicomponent diffusion behavior. The fractions of fast and slow apparent diffusion components have often been attributed to the volume fractions of extracellular space (ECS) and intracellular space (ICS) although diffusion fractions are at variance with the tissue compartment volume ratios. In this article this puzzle was examined with a finite difference diffusion simulation model on the basis of optical images from sectioned rat spinal cord. Here the results show that assignment of fractions obtained from biexponential fits of fast and slow diffusion attenuation to ECS and ICS volume ratios is not correct. Rather, the observed multicomponent diffusion behavior is caused by motional restriction and limited intercompartmental water exchange in that at long diffusion times diffusion attenuation is shown to become monoexponential. Although the measured apparent diffusion fractions also depend on T2 relaxation time of water protons in the various compartments, the sensitivity to T2 is small and thus T2 differences are unlikely to explain the mismatch between apparent diffusion fractions and cellular volume fractions. PMID- 11870832 TI - Assessment of T1 and T2* effects in vivo and ex vivo using iron oxide nanoparticles in steady state--dependence on blood volume and water exchange. AB - Accurate knowledge of the relationship between contrast agent concentration and tissue relaxation is a critical requirement for quantitative assessment of tissue perfusion using contrast-enhanced MRI. In the present study, using a pig model, the relationship between steady-state blood concentration levels of an iron oxide nanoparticle with a hydrated diameter of 12 nm (NC100150 Injection) and changes in the transverse and longitudinal relaxation rates (1/T2* and 1/T1, respectively) in blood, muscle, and renal cortex was investigated at 1.5 T. Ex vivo measurements of 1/T2* and 1/T1 were additionally performed in whole pig blood spiked with different concentrations of the iron oxide nanoparticle. In renal cortex and muscle, 1/T2* increased linearly with contrast agent concentration with slopes of 101 +/-22 s(-1)mM(-1) and 6.5 +/-0.9 s(-1)mM(-1) (mean +/- SD), respectively. In blood, 1/T2* increased as a quadratic function of contrast agent concentration, with different quadratic terms in the ex vivo vs. the in vivo experiments. In vivo, 1/T1 in blood increased linearly with contrast agent concentration, with a slope (T1-relaxivity) of 13.9 +/- 0.9 s(-1)mM(-1). The achievable increase in 1/T1 in renal cortex and muscle was limited by the rate of water exchange between the intra- and extravascular compartments and the 1/T1-curves were well described by a two-compartment water exchange limited relaxation model. PMID- 11870833 TI - Why does MTR change with neuronal depolarization? AB - T1 and T2 relaxation, and magnetization transfer (MT) of the rat brain were measured during experimentally induced spreading depression (SD). All measured MR parameters changed during SD: T1 relaxation increased by approximately 13%, whereas the T2 increase was substantially larger (88%). MT results showed an MT ratio (MTR) decrease of 9%. The lack of change in the MT exchange rate indicated that the MT processes between water and macromolecular protons are not affected by neuronal depolarization. The observed decrease in MTR was only caused by changes in T1 and T2 relaxation. PMID- 11870834 TI - Coronary artery motion with the respiratory cycle during breath-holding and free breathing: implications for slice-followed coronary artery imaging. AB - The displacement of the right coronary artery (RCA) origin with respiratory position was determined relative to the dome of the right hemidiaphragm in three orthogonal directions in eight healthy subjects. Both multiple breath-hold and free-breathing acquisitions were used, and motion correction factors for slice following applications were determined. The correction factors for all three directions showed considerable intersubject variability. The mean superior inferior factor was slightly less in free-breathing than in breath-holding (0.26 vs. 0.29, P = ns), and much less than the fixed value of 0.6 frequently implemented with slice-following. The anterior-posterior correction factors were uniformly low in free-breathing, and significantly less than those obtained from breath-holding (0.04 vs. 0.14, P <.05), while the mean left-right correction factors were approximately 0.1 for both. It is concluded that subject variability in correction factors, together with within-subject differences between breath holding and free-breathing, is such that slice-following should be performed with subject-specific factors determined from free-breathing acquisitions. PMID- 11870835 TI - Multislice first-pass cardiac perfusion MRI: validation in a model of myocardial infarction. AB - The purpose of this study was to validate a first-pass MRI method for imaging myocardial perfusion with multislice coverage and relatively small analyzable regions of interest (ROIs). A fast gradient-echo (FGRE) sequence with an echo train (ET) readout was used to achieve multislice coverage, and a high dose of a contrast agent (CA) was used to achieve a high signal-to-noise ratio (SNR). Dogs (N = 6) were studied 1 day after reperfused myocardial infarction, and fluorescent microspheres were used as a standard for perfusion. First-pass MRI correlated well vs. microsphere flow, achieving mean R values of 0.87 (range = 0.82-0.93), 0.71 (range = 0.46-0.85), and 0.72 (range = 0.49-0.95) for subendocardial ROIs, transmural ROIs, and the endocardial-epicardial ratio, respectively. Additionally, analysis of myocardial time-intensity curves (TICs) indicated that 15.8 +/- 6 sectors, corresponding to 260 microl of endocardium, can be analyzed (R(2) > 0.95). PMID- 11870838 TI - Oscillating dual-equilibrium steady-state angiography. AB - A novel technique of generating noncontrast angiograms is presented. This method, called oscillating dual-equilibrium steady-state angiography (ODESSA), utilizes a modified steady-state free precession (SSFP) pulse sequence. The SSFP sequence is modified such that flowing material reaches a steady state which oscillates between two equilibrium values, while stationary material attains a single, nonoscillatory steady state. Subtraction of adjacent echoes results in large, uniform signal from all flowing spins and zero signal from stationary spins. Venous signal can be suppressed based on its reduced T2. ODESSA arterial signal is more than three times larger than that of traditional phase-contrast angiography (PCA) in the same scan time, and also compares favorably with other techniques of MR angiography (MRA). Pulse sequences are implemented in 2D, 3D, and volumetric-projection modes. Angiograms of the lower leg, generated in as few as 5 s, show high arterial signal-to-noise ratio (SNR) and full suppression of other tissues. PMID- 11870836 TI - Fast MRI of RF heating via phase difference mapping. AB - A method is presented for the rapid acquisition of temperature maps derived from phase difference maps. The temperature-dependent chemical shift coefficients (TDCSCs) of various concentrations of aqueous cobalt and dysprosium-based compounds were measured. The largest TDCSC calculated was for 100 mM DyEDTA, which had a TDCSC of -0.09 PPM/K; 160 mM CoCl2 had a TDCSC of -0.04 PPM/K. These temperature-dependent chemical shifts (TDCSs) result in phase changes in the MR signal with changing temperature. Agarose phantoms were constructed with each paramagnetic metal. A fast gradient-echo (FGRE) MR image was acquired to serve as the baseline image. A "test" MRI procedure was then performed on the phantom. Immediately afterwards, a second FGRE MR image was acquired, serving as the probing image. Proper image processing as a phase difference map between the probing image and the baseline image resulted in an image which quantitatively described the temperature increase of the phantom in response to a particular "test" imaging experiment. Applications of this technique in assessing the safety of pulse sequences and MR coils are discussed. PMID- 11870837 TI - High temporal resolution phase contrast MRI with multiecho acquisitions. AB - Velocity imaging with phase contrast (PC) MRI is a noninvasive tool for quantitative blood flow measurement in vivo. A shortcoming of conventional PC imaging is the reduction in temporal resolution as compared to the corresponding magnitude imaging. For the measurement of velocity in a single direction, the temporal resolution is halved because one must acquire two differentially flow encoded images for every PC image frame to subtract out non-velocity-related image phase information. In this study, a high temporal resolution PC technique which retains both the spatial resolution and breath-hold length of conventional magnitude imaging is presented. Improvement by a factor of 2 in the temporal resolution was achieved by acquiring the differentially flow-encoded images in separate breath-holds rather than interleaved within a single breath-hold. Additionally, a multiecho readout was incorporated into the PC experiment to acquire more views per unit time than is possible with the single gradient-echo technique. A total improvement in temporal resolution by approximately 5 times over conventional PC imaging was achieved. A complete set of images containing velocity data in all three directions was acquired in four breath-holds, with a temporal resolution of 11.2 ms and an in-plane spatial resolution of 2 mm x 2 mm. PMID- 11870839 TI - Fast proton spectroscopic imaging with high signal-to-noise ratio: spectroscopic RARE. AB - A new fast spectroscopic imaging (SI) method is presented which is based on spatial localization by the fast MRI method of rapid acquisition with relaxation enhancement (RARE) and encoding of the chemical shift information by shifting the position of a refocusing 180 pulse in a series of measurements. This method is termed spectroscopic RARE. In contrast to spectroscopic ultrafast low-angle RARE (U-FLARE), the formation of two echo families (odd and even) is suppressed by using a train of 180 RF pulses with an internal four-step phase cycle. By this means a high signal-to-noise ratio (SNR) per unit measurement time is obtained, because the separation of odd and even echoes, as well as dummy echoes to stabilize the echo amplitudes, is not needed anymore. The method is of particular interest for detecting signals of coupled spins, as effective homonuclear decoupling can be achieved by use of constant evolution time chemical shift encoding. The pulse sequence was implemented on a 4.7 T imaging system, tested on phantoms, and applied to the healthy rat brain in vivo. Spectroscopic RARE is particularly useful if T2* double less-than sign T2, which is typically fulfilled for in vivo proton SI measurements at high magnetic field strength. PMID- 11870840 TI - Self-calibrating parallel imaging with automatic coil sensitivity extraction. AB - Calibration of the spatial sensitivity functions of coil arrays is a crucial element in parallel magnetic resonance imaging (PMRI). The most common approach has been to measure coil sensitivities directly using one or more low-resolution images acquired before or after accelerated data acquisition. However, since it is difficult to ensure that the patient and coil array will be in exactly the same positions during both calibration scans and accelerated imaging, this approach can introduce sensitivity miscalibration errors into PMRI reconstructions. This work shows that it is possible to extract sensitivity calibration images directly from a fully sampled central region of a variable density k-space acquisition. These images have all the features of traditional PMRI sensitivity calibrations and therefore may be used for any PMRI reconstruction technique without modification. Because these calibration data are acquired simultaneously with the data to be reconstructed, errors due to sensitivity miscalibration are eliminated. In vivo implementations of self calibrating parallel imaging using a flexible coil array are demonstrated in abdominal imaging and in real-time cardiac imaging studies. PMID- 11870841 TI - Combination of signals from array coils using image-based estimation of coil sensitivity profiles. AB - It is well established that the optimal unbiased way to combine image data from array coils is a pixel-by-pixel sum of coil signals, with each signal weighted by the individual coil sensitivity at the location of the pixel. A pragmatic alternative combines the images from the coils as the square root of the sum of squares (SOS), which can reduce the signal-to-noise ratio (SNR) and introduce bias. This work describes how to replace coil sensitivity by an image-derived quantity that enables close to optimal signal combination up to a global intensity scaling. Typical scaling is by an individual coil sensitivity or a linear or SOS combination of the sensitivities of some or all of the coils in the array. The method decreases signal bias, improves SNR when coils have unequal noise levels, and can reduce image artifacts. It can produce phase-corrected data, which eliminates bias completely. In addition, the method allows images from arrays that include highly localized coils, such as a prostate coil and external pelvic array, to be combined with near-optimal SNR and an intensity modulation that makes them easier to view. PMID- 11870842 TI - Assessment of a potential tumor-seeking manganese metalloporphyrin contrast agent in a mouse model. AB - The performance of a newly developed potential tumor-seeking magnetic resonance (MR) contrast agent alpha-Aqua-13,17-bis(1-carboxypropionyl) carbamoylethyl-3,8 bis(1-phenethyloxyethyl)-beta-hydroxy-2,7,12,18-tetramethyl-porphyrinato manganese (III) (HOP-8P) was tested using a mouse model. Tumor-bearing (SCC-VII) mice were imaged using a 1.5T MR imager before and after intravenous administration of 0.1 mmol/kg of HOP-8P. A biodistribution analysis was performed using an optical emission spectrometer. Significant enhancement of the transplanted tumor was observed in MR images 24 h after intravenous injection of HOP-8P. The biodistribution assessment of manganese also correlated with the results of the imaging study. During the 24-h period following contrast administration, HOP-8P was consistently cleared from the circulation, liver, kidneys, and muscle; however, it was progressively accumulated within the tumor. HOP-8P is a promising tumor-seeking metalloporphyrin MR contrast agent with a wide imaging window. PMID- 11870843 TI - Surface size, curvature analysis, and assessment of knee joint incongruity with MRI in vivo. AB - The purpose of this study was to develop an MR-based technique for quantitative analysis of joint surface size, surface curvature, and joint incongruity and to assess its reproducibility under in vivo imaging conditions. The surface areas were determined after 3D reconstruction of the joint by triangulation and the incongruity by Gaussian curvature analysis. The precision was tested by analyzing four replicated MRI datasets of human knees in 14 individuals. The algorithms were shown to produce accurate data in geometric test objects. The interscan precision was <4% (CV%) for surface area, 2.9-5.7 m(-1) (SD) for the mean principal curvature, and 4.1-7.4 m(-1) for congruence indices. Incongruity was highest in the femoropatellar joint (79.7 m(-1)) and lowest in the medial femorotibial joint (28.6 m(-1)). This technique will permit identification of the specific role of surface size, curvature, and incongruity as potential risk factors for osteoarthritis. PMID- 11870844 TI - ADC characterization of region-specific response to cerebral perfusion deficit in rats by MRI at 9.4 T. AB - Region-specific cerebral blood flow (CBF) and apparent diffusion coefficient (ADC) of water in the rat brain were quantified in vivo by high-field MRI (9.4 T) for 6-7 h after middle cerebral artery occlusion (MCAO). Upon occlusion, average CBF fell from about 1.5-2 ml/g/min to below 0.5 ml/g/min in cortical areas and the amygdala, and below 0.2 ml/g/min in the caudate putamen. CBF in some of the homologous contralateral areas also decreased by 20-30%. Average ADC decreased from about 8 center dot 10(-4) to 5 center dot 10(-4) mm(2)/s in the caudate putamen and parietal cortex. Corresponding changes in ADC were lower in the frontal cortex and negligible in the piriform cortex, suggesting that the perfusion threshold for ADC decrease may be different for different brain regions in the same animal. The area of decreased ADC correlated well with the infarction area revealed by 2,3,5-triphenyltetrazolium chloride (TTC) staining of brain slices in vitro. A better understanding of the mechanisms linking ADC and CBF changes to ischemic cell disorders may prove useful in characterizing the degree of tissue damage, and in developing and evaluating treatment strategies. PMID- 11870846 TI - Coupled microstrip line transverse electromagnetic resonator model for high-field magnetic resonance imaging. AB - The performance modeling of RF resonators at high magnetic fields of 4.7 T and more requires a physical approach that goes beyond conventional lumped circuit concepts. The treatment of voltages and currents as variables in time and space leads to a coupled transmission line model, whereby the electric and magnetic fields are assumed static in planes orthogonal to the length of the resonator, but wave-like along its longitudinal axis. In this work a multiconductor transmission line (MTL) model is developed and successfully applied to analyze a 12-element unloaded and loaded microstrip line transverse electromagnetic (TEM) resonator coil for animal studies. The loading involves a homogeneous cylindrical dielectric insert of variable radius and length. This model formulation is capable of estimating the resonance spectrum, field distributions, and certain types of losses in the coil, while requiring only modest computational resources. The boundary element method is adopted to compute all relevant transmission line parameters needed to set up the transmission line matrices. Both the theoretical basis and its engineering implementation are discussed and the resulting model predictions are placed in context with measurements. A comparison between a conventional lumped circuit model and this distributed formulation is conducted, showing significant departures in the resonance response at higher frequencies. This MTL model is applied to simulate two small-bore animal systems: one of 7.5 cm inner diameter, tuned to 200 MHz (4.7 T for proton imaging), and one of 13.36 cm inner diameter, tuned to both 200 and 300 MHz (7 T). PMID- 11870845 TI - EPR/NMR co-imaging for anatomic registration of free-radical images. AB - While electron paramagnetic resonance imaging (EPRI) enables spatial mapping of free radicals in the whole body of small animals, it solely visualizes the free radical distribution and does not typically provide anatomic visualization of the body. However, anatomic registration is often required for meaningful interpretation of the EPRI-derived free-radical images. An approach is reported for whole-body, EPRI-based, free-radical imaging along with proton MRI in mice. EPRI instrumentation with a 750-MHz narrow band microwave bridge and transverse oriented electric field reentrant resonator, with automatic coupling control and automatic tuning control capability, was used to map the spatial distribution of nitroxide free radicals in phantoms and in living mice, while low-field proton MRI at 16 MHz was used to define the anatomic structure to register the EPR images. Small capillary tubes containing an aqueous radical label were used as markers to enable image superimposition. With this coregistration technique, the EPRI free-radical images were precisely registered, enabling assignment of the location of the observed free-radical distribution within the organs of the mice. This technique enabled differentiation of the distribution and metabolism of nitroxide radicals within the major organs and body sites of living mice. PMID- 11870848 TI - Uncertainty in the analysis of tracer kinetics using dynamic contrast-enhanced T1 weighted MRI. AB - In recent years a number of physiological models have gained prominence in the analysis of dynamic contrast-enhanced T1-weighted MRI data. However, there remains little evidence to support their use in estimating the absolute values of tissue physiological parameters such as perfusion, capillary permeability, and blood volume. In an attempt to address this issue, data were simulated using a distributed pathway model of tracer kinetics, and three published models were fitted to the resultant concentration-time curves. Parameter estimates obtained from these fits were compared with the parameters used for the simulations. The results indicate that the use of commonly accepted models leads to systematic overestimation of the transfer constant, Ktrans, and potentially large underestimates of the blood plasma volume fraction, Vp. In summary, proposals for a practical approach to physiological modeling using MRI data are outlined. PMID- 11870847 TI - Multifunctional interventional devices for MRI: a combined electrophysiology/MRI catheter. AB - The design and application of a two-wire electrophysiology (EP) catheter that simultaneously records the intracardiac electrogram and receives the MR signal for active catheter tracking is described. The catheter acts as a long loop receiver, allowing for visualization of the entire catheter length while simultaneously behaving as a traditional two-wire EP catheter, allowing for intracardiac electrogram recording and ablation. The application of the device is demonstrated by simultaneously tracking the catheter and recording the intracardiac electrogram in canine models using 7 and 10 frame/sec real-time imaging sequences. Using solely MR imaging, the entire catheter was visualized and guided from the jugular vein into the cardiac chambers, where the intracardiac electrogram was recorded. By combining several functions in a single, simple structure, the excellent tissue contrast and functional imaging capabilities of MR can be used to improve the efficacy of EP interventions. This catheter will facilitate MR-guided interventions and demonstrates the design of multifunctional interventional devices for use in MRI. PMID- 11870849 TI - Bulk magnetic susceptibility effects on the assessment of intra- and extramyocellular lipids in vivo. AB - Localized proton spectroscopy provides a novel method for noninvasive measurement of lipid content in skeletal muscle. It has been suggested that the chemical shift difference between lipid signals from distinct compartments in skeletal muscle might be caused by bulk magnetic susceptibility (BMS) differences from lipids stored in intra- (IMCL) and extramyocellular (EMCL) compartments. Direct evidence is provided to confirm the theoretical prediction that compartment symmetry is responsible for discrimination between resonances of IMCL and EMCL. Phantoms imitating lipids in skeletal muscle were constructed using soybean oil to represent EMCL, and IntralipidTM, an intravenous fat emulsion of fine droplets, to represent IMCL. It was found that the chemical shift of IntralipidTM is independent of the BMS effects, while the resonance of soybean oil shifts in a predictable manner determined by the geometry of the compartment. PMID- 11870850 TI - Half-Fourier single-shot STEAM MRI. AB - As a high-speed imaging technique based on stimulated echoes single-shot STEAM MRI is insensitive to chemical shift artifacts and magnetic susceptibility differences. The achievable signal-to-noise ratio (SNR) is limited by the fact that high flip angles of the read-out excitation pulses cause a steep decay of the stimulated echo train and therefore degrade the point-spread function of the resulting image. The present work investigates the use of half-Fourier phase encoding which weakens the flip angle constraint by reducing the number of necessary excitations. Single-shot STEAM MRI of the normal human brain at 2.0 T demonstrates that half-Fourier versions either reduce the measurement time by almost a factor of two without sacrificing the SNR or increase the SNR by about 40% while keeping the measurement time constant. PMID- 11870851 TI - Multislice double inversion pulse sequence for efficient black-blood MRI. AB - Over the last several years there has been a rapidly growing interest in high resolution MRI of the vascular wall to assess the extent of atherosclerotic lesions. Vessels of particular clinical relevance are the carotid and coronary arteries. Currently, the preferred imaging sequence for these studies is a "black blood" technique based on the double-inversion scheme to null the blood signal. A critical drawback of the black-blood technique, however, has been its single slice nature, as there is only one point in time during the recovery of the blood magnetization from inversion at which the signal is completely nulled. Consequently, the total scan time can become prohibitively long, particularly when an imaging protocol includes several series of these datasets. In this work, a multiple-slice double-inversion technique is described that can reduce the scan time by a factor of two or more. It is demonstrated in vivo with examples from carotid and coronary arteries that one can acquire multiple slices with sufficient nulling of blood, following a single set of inversion pulses. PMID- 11870854 TI - ABC transporters Cdr1p, Cdr2p and Cdr3p of a human pathogen Candida albicans are general phospholipid translocators. AB - We have used fluorescent 7-nitrobenz-2-oxa-1,3-diazol-4-yl (NBD)-tagged phospholipid analogues, NBD-PE (phosphatidylethanolamine), NBD-PC (phosphatidylcholine) and NBD-PS (phosphatidylserine), to demonstrate that Cdr1p and its other homologues, Cdr2p and Cdr3p, belonging to the ATP-binding cassette (ABC) superfamily behave as general phospholipid translocators. Interestingly, CDR1 and CDR2, whose overexpression leads to azole resistance in C. albicans, elicit in-to-out transbilayer phospholipid movement, while CDR3, which is not involved in drug resistance, carries out-to-in translocation of phospholipids between the two monolayers of plasma membrane. Cdr1p, Cdr2p and Cdr3p could be further distinguished on the basis of their sensitivities to different inhibitors. For example, the in-to-out activity associated with Cdr1p and Cdr2p is energy-dependent and sensitive to sulphydryl blocking agents such as N ethylmaleimide (NEM) and cytoskeleton disrupting agent cytochalasin E, while Cdr3p-associated out-to-in activity is energy-dependent but insensitive to NEM and cytochalasin E. We found that certain drugs, such as fluconazole, cycloheximide and miconazole, to which Cdr1p confers resistance could also affect in-to-out transbilayer movement of NBD-PE, while the same drugs had no effect on Cdr3p-mediated out-to-in translocation of NBD-PE. The ineffectiveness of these drugs to affect Cdr3p mediated out-to-in phospholipid translocation further confirms the inherent difference in the directionality of phospholipid translocation between these pumps. Notwithstanding the role of some of the Cdrps in drug resistance, this study clearly demonstrates that these ABC transporters of C. albicans are phospholipid translocators and this function could represent one of the physiological functions of such large family of proteins. PMID- 11870853 TI - Decreasing acetic acid accumulation by a glycerol overproducing strain of Saccharomyces cerevisiae by deleting the ALD6 aldehyde dehydrogenase gene. AB - Glycerol is a major fermentation product of Saccharomyces cerevisiae that contributes to the sensory character of wine. Diverting sugar to glycerol overproduction and away from ethanol production by overexpressing the glycerol 3 phosphate dehydrogenase gene,GPD2, caused S. cerevisiae to produce more than twice as much acetic acid as the wild-type strain (S288C background) in anaerobic cell culture. Deletion of the aldehyde dehydrogenase gene, ALD6, in wild-type and GPD2 overexpressing strains (GPD2-OP) decreased acetic acid production by three- and four-fold, respectively. In conjunction with reduced acetic acid production, the GPD2-OP ald6Delta strain produced more glycerol and less ethanol than the wild-type. The growth rate and fermentation rate were similar for the modified and wild-type strains, although the fermentation rate for the GPD2 ald6Delta strain was slightly less than that of the other strains from 24h onwards. Analysis of the metabolome of the mutants revealed that genetic modification affected the production of some secondary metabolites of fermentation, including acids, esters, aldehydes and higher alcohols, many of which are flavour-active in wine. Modification of GPD2 and ALD6 expression represents an effective strategy to increase the glycerol and decrease the ethanol concentration during fermentation, and alters the chemical composition of the medium such that, potentially, novel flavour diversity is possible. The implications for the use of these modifications in commercial wine production require further investigation in wine yeast strains. PMID- 11870855 TI - Efficient PCR-based gene disruption in Saccharomyces strains using intergenic primers. AB - Gene disruptions are a vital tool for understanding Saccharomyces cerevisiae gene function. An arrayed library of gene disruption strains has been produced by a consortium of yeast laboratories; however their use is limited to a single genetic background. Since the yeast research community works with several different strain backgrounds, disruption libraries in other common laboratory strains are desirable. We have developed simple PCR-based methods that allow transfer of gene disruptions from the S288C-derived strain library into any Saccharomyces strain. One method transfers the unique sequence tags that flank each of the disrupted genes and replaces the kanamycin resistance marker with a recyclable URA3 gene from Kluyveromyces lactis. All gene-specific PCR amplifications for this method are performed using a pre-existing set of primers that are commercially available. We have also extended this PCR technique to develop a second general gene disruption method suitable for any transformable strain of Saccharomyces. PMID- 11870856 TI - Siderophore uptake by Candida albicans: effect of serum treatment and comparison with Saccharomyces cerevisiae. AB - Iron uptake systems often function as virulence factors in pathogenic organisms. Candida albicans is a fungal pathogen that infects immunocompromised hosts, such as AIDS patients or granulocytopenic bone marrow transplant recipients. Here we show that iron uptake from siderophores occurs in C. albicans and is mediated by one or more high-affinity transport systems. Iron carried on ferrioxamine B, triacethyl-fusarinine, ferrichrome, or ferricrocin was actively taken up via a high-affinity mechanism. The kinetic parameters of uptake were similar to those found in S. cerevisiae. Furthermore, for ferrichrome and ferrioxamine B, cellular uptake of fluorescent analogues was observed. In C. albicans, iron uptake from siderophores was regulated by iron availability, with iron deprivation inducing uptake. Serum exposure, which induces a morphogenic shift from yeast to filamentous forms known to be required for virulence, also resulted in induction of iron transport from ferrichrome-type siderophores. In a tup1/tup1 strain which grows constitutively in the filamentous form, iron transport was derepressed for all siderophores tested. The genes mediating uptake and utilization of iron from siderophores in C. albicans have not been identified; however, the transcript abundance for CaSIT1 was regulated in a manner consistent with the pattern of iron uptake from ferrichrome-type siderophores. Furthermore, CaSIT1 overexpression in S. cerevisiae resulted in inhibited siderophore iron uptake, suggesting that the expressed protein may interact with proteins of S. cerevisiae involved in iron uptake from siderophores. In summary, iron uptake from ferrichrome-type siderophores was induced in filamentous C. albicans, and a potential role of this iron acquisition system in pathogenicity should be considered. PMID- 11870857 TI - Inhibitory effect of nikkomycin Z on chitin synthases in Candida albicans. AB - Nikkomycin Z is a competitive inhibitor of chitin synthases in fungi. It has been reported that it inhibits chitin synthases (Chs) 1 and 3, but not 2, of Saccharomyces cerevisiae. In our study, we found that: (a) nikkomycin Z inhibited all three Chs isozymes of Candida albicans (CaChs). The IC(50) value for CaChs1 is 15microM, for CaChs2 0.8microM, and for CaChs3 13microM; (b) nikkomycin Z inhibits vegetative growth of C. albicans differently in different growth media; growth inhibition was observed on Spider and corn meal agar plate, but not on Lee's plate; (c) growth inhibition by nikkomycin Z accompanied by the absence of septum and cell wall chitin, which in turn brought about cell lysis. Nikkomycin Z did not lyse cells in Lee's media and lysis was partially prevented in the presence of sorbitol as an osmostabilizer in Spider medium. Therefore, nikkomycin Z prevented the formation of septum and cell wall chitin by inhibiting chitin synthase activities in a growth medium-dependent manner. PMID- 11870858 TI - Mutants defective in secretory/vacuolar pathways in the EUROFAN collection of yeast disruptants. AB - We have screened the EUROFAN (European Functional Analysis Network) deletion strain collection for yeast mutants defective in secretory/vacuolar pathways and/or associated biochemical modifications. We used systematic Western immunoblotting to analyse the electrophoretic pattern of several markers of the secretory/vacuolar pathways, the soluble alpha-factor, the periplasmic glycoprotein invertase, the plasma membrane GPI-anchored protein Gas1p, and two vacuolar proteins, the soluble carboxypeptidase Y and the membrane-bound alkaline phosphatase, which are targeted to the vacuole by different pathways. We also used colony immunoblotting to monitor the secretion of carboxypeptidase Y into the medium, to identify disruptants impaired in vacuolar targeting. We identified 25 mutants among the 631 deletion strains. Nine of these mutants were disrupted in genes identified in recent years on the basis of their involvement in trafficking (VPS53, VAC7, VAM6, APM3, SYS1), or glycosylation (ALG12, ALG9, OST4, ROT2). Three of these genes were identified on the basis of trafficking defects by ourselves and others within the EUROFAN project (TLG2, RCY1, MON2). The deletion of ERV29, which encodes a COPII vesicle protein, impaired carboxypeptidase Y trafficking from the endoplasmic reticulum to the Golgi apparatus. We also identified eight unknown ORFs, the deletion of which reduced Golgi glycosylation or impaired the Golgi to vacuole trafficking of carboxypeptidase Y. YJR044c, which we identified as a new VPS gene, encodes a protein with numerous homologues of unknown function in sequence databases. PMID- 11870859 TI - Proliferation of the endoplasmic reticulum occurs normally in cells that lack a functional unfolded protein response. AB - Increased expression of certain ER membrane proteins leads to biogenesis of novel ER membrane arrays. These structures provide models in which to explore the mechanisms by which cells control the size and organization of organelles in response to changing physiological demands. In yeast, elevated levels of HMG-CoA reductase induce ER arrays known as karmellae. Cox and co-workers (1997) discovered that karmellae assembly is toxic to ire1 mutants. These mutants are unable to initiate the unfolded protein response, which enables cells to adjust levels of ER chaperones in response to stresses. We sought to determine whether the karmellae-dependent death of ire1 mutants was due to karmellae assembly or to increased levels of HMG-CoA reductase activity. Unexpectedly, we found that ire1 cells could assemble normal levels of karmellae that were structurally identical to those of wild-type cells. In addition, karmellae assembly did not itself induce the unfolded protein response. Certain ire1 strains produced significant numbers of transformants that were unable to utilize galactose as sole carbon source. These results suggest that the karmellae-dependent death of certain ire1 strains may simply reflect their inability to grow on galactose. PMID- 11870860 TI - Geometric and chemical patterns of interaction in protein--ligand complexes and their application in docking. AB - We present a new method for representing the binding site of a protein receptor that allows the use of the DOCK approach to screen large ensembles of receptor conformations for ligand binding. The site points are constructed from templates of what we called "attached points" (ATPTS). Each template (one for each type of amino acid) is composed of a set of representative points that are attached to side-chain and backbone atoms through internal coordinates, carry chemical information about their parent atoms and are intended to cover positions that might be occupied by ligand atoms when complexed to the protein. This method is completely automatic and proved to be extremely fast. With the aim of obtaining an experimental basis for this approach, the Protein Data Bank was searched for proteins in complex with small molecules, to study the geometry of the interactions between the different types of protein residues and the different types of ligand atoms. As a result, well-defined patterns of interaction were obtained for most amino acids. These patterns were then used for constructing a set of templates of attached points, which constitute the core of the ATPTS approach. The quality of the ATPTS representation was demonstrated by using this method, in combination with the DOCK matching and orientation algorithms, to generate correct ligand orientations for >1000 protein--ligand complexes. PMID- 11870861 TI - Solution structures of a 30-residue amino-terminal domain of the carp granulin-1 protein and its amino-terminally truncated 3-30 subfragment: implications for the conformational stability of the stack of two beta-hairpins. AB - Carp granulins are members of an emerging class of proteins with a sequence motif encoding a parallel stack of two to four beta-hairpins. The carp granulin-1 protein forms a stack of four beta-hairpins, whereas its amino-terminal fragment appears to adopt a very stable stack of two beta-hairpins in solution. Here we determined a refined three-dimensional structure of this peptide fragment to examine potential conformational changes compared with the full-length protein. The structures were calculated with both a traditional method and a fast semiautomated method using ambiguous NMR distance restraints. The resulting sets of structures are very similar and show that a well-defined stack of two beta hairpins is retained in the peptide. Conformational rearrangements compensating the loss of the carboxy-terminal subdomain of the native protein are restricted to the carboxy-terminal end of the peptide, the turn connecting the two beta hairpins, and the Tyr(21) and Tyr(25) aromatic side chains. Further removal of the Val(1) and Ile(2) residues, which are part of the first beta-hairpin and components of two major hydrophobic clusters in the two beta-hairpin structure, results in the loss of the first beta-hairpin. The second beta-hairpin, which is closely associated with the first, retains a similar but somewhat less stable conformation. The invariable presence of the second beta-hairpin and the dependence of its stability on the first beta-hairpin suggest that the stack of two beta-hairpins may be an evolutionary conserved and autonomous folding unit. In addition, the high conformational stability makes the stack of two beta hairpins an attractive scaffold for the development of peptide-based drug candidates. PMID- 11870862 TI - Congruent qualitative behavior of complete and reconstructed phase space trajectories from biomolecular dynamics simulation. AB - Central to the study of a complex dynamical system is knowledge of its phase space behavior. Experimentally, it is rarely possible to record a system's (multidimensional) phase space variables. Rather, the system is observed via one (or few) scalar-valued signal(s) of emission or response. In dynamical systems analysis, the multidimensional phase space of a system can be reconstructed by manipulation of a one-dimensional signal. The trick is in the construction of a (higher-dimensional) space through the use of a time lag (or delay) on the signal time series. The trajectory in this embedding space can then be examined using phase portraits generated in selected subspaces. By contrast, in computer simulation, one has an embarrassment of riches: direct access to the complete multidimensional phase space variables, at arbitrary time resolution and precision. Here, the problem is one of reducing the dimensionality to make analysis tractable. This can be achieved through linear or nonlinear projection of the trajectory into subspaces containing high information content. This study considers trajectories of the small protein crambin from molecular dynamics simulations. The phase space behavior is examined using principal component analysis on the Cartesian coordinate covariance matrix of 138 dimensions. In addition, the phase space is reconstructed from a one dimensional signal, representing the radius of gyration of the structure along the trajectory. Comparison of low-dimensional phase portraits obtained from the two methods shows that the complete phase space distribution is well represented by the reconstruction. The study suggests that it may be possible to develop a deeper connection between the experimental and simulated dynamics of biomolecules via phase space reconstruction using data emerging from recent advances in single molecule time-resolved biophysical techniques. PMID- 11870863 TI - A clade of trypsins found in cold-adapted fish. AB - A clade of trypsins, known as group III, is identified by phylogenetic analysis. These trypsins occur in fish that spend all or part of their lives at temperatures near 0 degrees C and may represent extreme psychrophilic enzymes. A principal component analysis of amino acid compositions distinguishes group III from mesophilic trypsins, as do molecular trees and multidimensional scaling of molecular sequence distances. The primary sequences of group III trypsins, in conjunction with the known structures of mesophilic trypsins, permit insight into function and mechanisms of cold adaption. The techniques employed are broadly applicable to phylogenies characterized by a markedly different, or "fast evolving," clade. An updated lactate dehydrogenase molecular tree illustrates an additional fast-evolving clade. PMID- 11870864 TI - A critical analysis of continuum electrostatics: the screened Coulomb potential- implicit solvent model and the study of the alanine dipeptide and discrimination of misfolded structures of proteins. AB - An analysis of the screened Coulomb potential--implicit solvent model (SCP--ISM) is presented showing that general equations for both the electrostatic and solvation free energy can be derived in a continuum approach, using statistical averaging of the polarization field created by the solvent around the molecule. The derivation clearly shows how the concept of boundary, usually found in macroscopic approaches, is eliminated when the continuum model is obtained from a microscopic treatment using appropriate averaging techniques. The model is used to study the alanine dipeptide in aqueous solution, as well as the discrimination of native protein structures from misfolded conformations. For the alanine dipeptide the free energy surface in the phi--psi space is calculated and compared with recently reported results of a detailed molecular dynamics simulation using an explicit representation of the solvent, and with other available data. The study showed that the results obtained using the SCP--ISM are comparable to those of the explicit water calculation and compares favorably to the FDPB approach. Both transition states and energy minima show a high correlation (r > 0.98) with the results obtained in the explicit water analysis. The study of the misfolded structures of proteins comprised the analysis of three standard decoy sets, namely, the EMBL, Park and Levitt, and Baker's CASP3 sets. In all cases the SCP--ISM discriminated well the native structures of the proteins, and the best-predicted structures were always near-native (cRMSD approximately 2 A). PMID- 11870865 TI - NADH interactions with WT- and S94A-acyl carrier protein reductase from Mycobacterium tuberculosis: an ab initio study. AB - We present an ab initio molecular dynamics study of the complex between acyl carrier protein reductase InhA from M. tuberculosis and isonicotinic acid hydrazide-NADH. We focus on wild-type (WT) InhA and a mutant causing drug resistance (S94A) for which structural information is available (Rozwarski et al., 1998;279:98--102; Dessen et al., 1995;267:1638--1641). Our calculations suggest that the water-mediated H-bond interactions between Ser94 side chain and NADH, present in WT InhA X-ray structure, can be lost during the dynamics. This conformational change is accompanied by a structural rearrangement of Gly14. The calculated structure of WT is rather similar to the X-ray structure of the S94A mutant in terms of geometrical parameters and chemical bonding. Further evidence for the mobility of Ser94 is provided by a 1-ns-long classical molecular dynamics on the entire protein. The previously unrecognized high mobility of Ser94 can provide a rationale of the small change in free binding energies on passing from WT to S94A InhA. PMID- 11870866 TI - Refining structural and functional predictions for secretasome components by comparative sequence analysis. AB - Comparative sequence analysis of presenilins reveals the conserved transmembrane domain shared with leukocyte antigen CD47, possibly involved in signal transduction. Sensitive techniques of multiple sequence alignment extend the earlier observation of the aminopeptidase homology domain in nicastrin to suggest that this protein may be a catalytically active component of secretasome involved in proteolysis or co-proteolysis of presenilin or beta-amyloid. PMID- 11870867 TI - Protein--protein docking of electron transfer complexes: cytochrome c oxidase and cytochrome c. AB - Electron transferring protein complexes form only transiently and the crystal structures of electron transfer protein--protein complexes involving cytochrome c could so far be determined only for the pairs of yeast cytochrome c peroxidase (CcP) with iso-1-cytochrome c (iso-1-cyt c) and with horse heart cytochrome c (cyt c). This article presents models from computational docking for complexes of cytochrome c oxidase (COX) from Paracoccus denitrificans with horse heart cytochrome c, and with its physiological counterpart cytochrome c552 (c552). Initial docking is performed with the FTDOCK program, which permits an exhaustive search of translational and rotational space. A filtering procedure is then applied to reduce the number of complexes to a manageable number. In a final step of structural and energetic refinement, the complexes are optimized by rigid-body energy minimization with the molecular mechanics package CHARMM. This methodology was first tested on the CcP:iso-1-cyt c complex, in which the complex with the lowest CHARMM energy has an RMSD from the crystal structure of only 1.8 A (C(alpha) carbon atoms). Notably, the crystal conformation has an even lower energy. The same procedure was then applied to COX:cyt c and COX:c552. The lowest energy COX:cyt c complex is very similar to a docking model previously described for the complex of bovine cytochrome c oxidase with horse heart cytochrome c. For the COX:c552 complex, cytochrome c552 is found in two different orientations, depending on whether it is docked against COX from a two-subunit or from a four subunit crystal structure, respectively. Both conformations are discussed critically in the light of the available experimental data. PMID- 11870868 TI - Noncovalent interaction energies in covalent complexes: TEM-1 beta-lactamase and beta-lactams. AB - The class A beta-lactamase TEM-1 is a key bacterial resistance enzyme against beta-lactam antibiotics, but little is known about the energetic bases for complementarity between TEM-1 and its inhibitors. Most inhibitors form a covalent adduct with the catalytic Ser70, making the measurement of equilibrium constants, and hence interaction energies, technically difficult. This study evaluates noncovalent interactions within covalent complexes by examining the differential stability of TEM-1 and its inhibitor adducts. The thermal denaturation of TEM-1 follows a two-state, reversible model with a melting temperature (T(m)) of 51.6C and a van't Hoff enthalpy of unfolding (DeltaH(VH)) of 146.2 kcal/mol at pH 7.0. The stability of the enzyme changes on forming an inhibitor adduct. As expected, some inhibitors stabilize TEM-1; transition-state analogues increase the T(m) by up to 3.7C (1.7 kcal/mol). Surprisingly, all beta-lactam covalent acyl--enzyme complexes tested destabilize TEM-1 significantly relative to the apo-enzyme. For instance, the clinically used inhibitor clavulanic acid and the beta-lactamase resistant beta-lactams moxalactam and imipenem destabilize TEM-1 by over 2.6C (1.2 kcal/mol) in their covalent adducts. Based on the structure of the TEM 1/imipenem complex (Maveyraud et al., J Am Chem Soc 1998;120:9748--52), destabilization by moxalactam and imipenem is thought to be caused by a steric clash between the side-chain of Asn132 and the 6(7)-alpha group of these beta lactams. To test this hypothesis, the mutant enzyme N132A was made. In contrast with wild-type, the covalent complexes between N132A and both imipenem and moxalactam stabilize the enzyme, consistent with the hypothesis. To investigate the structural bases of this dramatic change in stability, the structure of N132A/imipenem was determined by X-ray crystallography. In the complex with N132A, imipenem adopts a very different conformation from that observed in the wild-type complex, and the putative destabilizing interaction with residue 132 is relieved. Studies of several enzymes suggest that beta-lactams, and covalent inhibitors in general, can have either net favorable or net unfavorable noncovalent interaction energies within the covalent complex. In the case of TEM 1, such unfavorable interactions convert substrate analogues into very effective inhibitors. PMID- 11870869 TI - Borna disease virus-induced accumulation of macrophage migration inhibitory factor in rat brain astrocytes is associated with inhibition of macrophage infiltration. AB - To test the hypothesis that macrophage migration inhibitory factor (MIF) plays a role in macrophage invasion during virus-induced encephalitis, we analyzed the expression and cellular localization of MIF in the Borna disease virus (BDV) infected rat brain, monitored monocyte/macrophage infiltration, and evaluated the influence of anti-inflammatory treatment with dexamethasone. MIF mRNA expression was restricted to neurons and remained unchanged after BDV infection or after dexamethasone treatment of either BDV-infected or uninfected control rats. In contrast, MIF protein immunoreactivity (ir) was not only seen in neurons but also in glia. After BDV-induced encephalitis and treatment of uninfected rats with dexamethasone, MIF ir was only slightly altered in neurons but moderately enhanced in tanycytes, ependyma, and choroid plexus epithelium and markedly increased or induced in astrocyte end-feet at the blood-brain barrier (BBB). The increase in MIF ir in astrocytes after BDV infection was blocked by dexamethasone. The induction or enhancement of MIF ir at the BBB significantly correlated with reduced numbers of infiltrating ED1-positive monocytes/macrophages after BDV infection. Increased macrophage invasion was observed in regions where no astrocytic MIF was detected. The BDV- or dexamethasone-induced accumulation of MIF protein in astrocytes in vivo in absence of detectable astrocytic MIF mRNA expression is most likely due to MIF translocation from neurons rather than to a constitutive or induced MIF mRNA expression in astrocytes. In conclusion, we provide evidence that translocation of MIF from neurons or other extracellular sources into astrocytes is likely to modulate the inflammatory process during the course of virus-induced encephalitis by limiting monocyte/macrophage migration through the BBB. PMID- 11870870 TI - Efficient recolonisation of progenitor-depleted areas of the CNS by adult oligodendrocyte progenitor cells. AB - A widely quoted hypothesis for the failure of remyelination in multiple sclerosis (MS) is the exhaustion of the oligodendrocyte progenitor cell (OPC) pool that is strongly implicated as the source of remyelinating oligodendrocytes in demyelinating lesions. Despite this, little is known about the responses of adult OPCs to adjacent areas of the CNS from which their numbers are depleted. We have developed an experimental model to study the pattern and rate of repopulation of OPC-depleted zones, by endogenous OPCs in the adult rat spinal cord. By X irradiating short lengths of the spinal cord with 40 Gy of X-irradiation, we were able to produce a highly localised depletion of OPCs that allowed us to study the responses of cells located in adjacent normal areas, to this local depletion. Using both NG2 immunohistochemistry and PDGFalphaR in situ hybridisation to identify OPCs, we demonstrate that endogenous OPCs repopulated the depleted areas slowly, but completely. This repopulation occurred at the rate of approximately 0.5 mm/week in the first month. Most cells at the leading edge of repopulation had complex, branching morphologies. The repopulation process was capable of restoring the density of progenitors in repopulated areas to that of normal tissue and was not associated with a secondary progenitor loss in tissue from which progenitor cells were generated. These findings indicate that depletion of the OPC population around lesions is not likely to be the primary explanation for remyelination failure in MS. PMID- 11870871 TI - Expression of fractalkine (CX3CL1) and its receptor, CX3CR1, during acute and chronic inflammation in the rodent CNS. AB - In this study, we investigate the expression of fractalkine (CX3CL1) and the fractalkine receptor (CX3CR1) in the naive rat and mouse central nervous system (CNS). We determine if the expression of this chemokine and its receptor are altered during chronic or acute inflammation in the CNS. In addition, we determine if CX3CL1, which has been reported to be chemoattractant to leukocytes in vitro, is capable of acting as a chemoattractant in the CNS in vivo. Immunohistochemistry was performed using primary antibodies recognizing soluble and membrane-bound CX3CL1 and the N-terminus of the CX3CR1. We found that neurons in the naive rodent brain are immunoreactive for CX3CL1 and CX3CR1, both showing a perinuclear staining pattern. Resident microglia associated with the parenchyma and macrophages in the meninges and choroid plexus constituitively express CX3CR1. In a prion model of chronic neurodegeneration and inflammation, CX3CL1 immunoreactivity is upregulated in astrocytes and CX3CR1 expression is elevated on microglia. In surviving neurons, expression of CX3CL1 appears unaltered relative to normal neurons. There is a decrease in neuronal CX3CR1 expression. Acute inflammatory responses in the CNS, induced by stereotaxic injections of lipopolysaccharide or kainic acid, results in activation of microglia and astrocytes but no detectable changes in the glial expression of CX3CL1 or CX3CR1. The expression of CX3CL1 and CX3CR1 by glial cells during inflammation in the CNS may be influenced by the surrounding cytokine milieu, which has been shown to differ in acute and chronic neuroinflammation. PMID- 11870872 TI - Preparation of a monoclonal antibody to citrullinated epitopes: its characterization and some applications to immunohistochemistry in human brain. AB - Using hybridoma technology, an IgM monoclonal antibody (mAb), designated as F95, was developed against a deca-citrullinated peptide (DCP) consisting of 10 citrulline residues and a carboxyl Gly-Gly-Cys through which DCP was covalently linked to an activated carrier protein, keyhole limpet hemocyanin (KLH). Clones were selected on the basis of not reacting with human unmodified and noncitrullinated myelin basic protein (MBP), MBP-C1, but reacting well with human citrullinated MBP (MBP-C8). When tested by ELISA, this mAb demonstrated minimal reactivity with human MBP-C1, varying reactivity with the C2-C5 isomers of human MBP, moderate binding with guinea pig MBP-C8, and strong reactivity with human MBP-C8. By ELISA, mAb F95 was directed predominantly against citrulline, not MBP, as revealed by its binding to DCP linked with activated KLH, bovine serum albumin (BSA), or ovalbumin (OA), but not with KLH, BSA, or OA alone. Immunohistochemistry of normal human brain demonstrated that F95 stained central nervous system myelin and a subset of astrocytes. Given the citrulline-directed features of mAb F95, this immunohistochemical pattern suggests that certain astroglial filaments expressing glial fibrillary acidic protein also contain citrulline-bearing components. These potentially implicate citrullinated proteins, notably in astroglial filaments, in a variety of normal and pathological neurobiological processes. PMID- 11870873 TI - Mts1 protein expression in the central nervous system after injury. AB - We recently showed that Mts1 is expressed in white matter astrocytes in the rat brain and spinal cord from the first postnatal day. Its expression level declined in the adult CNS, but its topographical localization was maintained. Only white matter astrocytes in the cerebellum did not express Mts1. After dorsal root or sciatic nerve injury, we observed a marked upregulation of Mts1 in the area of the dorsal funiculus undergoing Wallerian degeneration. Here we show that upregulation of Mts1 is a consistent feature of astrocytes in white matter undergoing Wallerian degeneration. In addition, Mts1 is upregulated in astrocytes outlining the lesion site of a penetrating injury to the forebrain, or cerebellum. Gray matter astrocytes did not express Mts1, even after direct injury. In injured brain, we consistently noted a close relationship between Mts1 expressing astrocytes and ED1-positive microglia/macrophages, which are known to be highly motile cells. Mts1 was expressed in the periventricular area and the rostral migratory stream, i.e., sites of ongoing neuroplasticity in adulthood, and was upregulated in these areas after injury. These data suggest that Mts1 expressing astrocytes play a significant role in degenerative events in the mature white matter, interact with phagocytic microglia/macrophages and regulate cell migration and differentiation in areas of the adult brain with a high degree of plasticity. PMID- 11870874 TI - Identification of growth factors that promote long-term proliferation of olfactory ensheathing cells and modulate their antigenic phenotype. AB - Olfactory ensheathing cells can develop into distinct subtypes in culture after incubation in serum-free medium conditioned by astrocytes, which have Schwann cell-like and astrocyte-like properties. It has not been possible so far to modulate and grow large numbers of these olfactory ensheathing cell subtypes. In this study, we have shown that astrocyte-conditioned medium, although promoting differentiation of the two olfactory ensheathing cell types, is growth restrictive after 14 days, probably due to the upregulation of p16 and p27. Growth arrest can be overridden and cells maintained for a further 11 weeks, by a mitogen mix of fibroblast growth factor 2, forskolin, and heregulin (olfactory mitogen medium) combined with astrocyte-conditioned medium. In the absence of astrocyte-conditioned medium, combinations of the same factors can also override growth arrest but to a lesser extent. Olfactory mitogen medium combined with astrocyte-conditioned medium upregulates O4 and low-affinity nerve growth factor receptor expression on olfactory ensheathing cells, leading to a 100% Schwann cell--like phenotype. If cells are maintained in olfactory mitogen medium alone, or if they are treated with forskolin or fibroblast growth factor 2 diluted in serum-free medium, O4 and low-affinity nerve growth factor receptor expression remains at 100%, but there is also an increase in expression of E-NCAM, the astrocyte-like marker. Medium containing serum also overrides growth arrest, but for only 4 weeks, during which time most differentiation-specific markers disappear. These studies have allowed us to define conditions to modulate the olfactory ensheathing cell phenotype. PMID- 11870875 TI - Functional expression of V-ATPases in the plasma membrane of glial cells. AB - Vacuolar H(+) ATPase (V-ATPase) activity is essential for many cellular processes, including intracellular membrane traffic, protein processing and degradation, and receptor-mediated endocytosis. Proton transport by V-ATPases could also play a role during cell transformation, tumorigenesis, and cell metastasis, and V-ATPase c-subunit overexpression was reported to be correlated with invasiveness of pancreatic tumors (Ohta et al., 1996). In the present work, we found that mRNAs encoding V-ATPase subunits are not overexpressed in C6 tumoral glioma cells when compared with immortalized astrocytes DI TNC1 and astrocytes in primary cultures. Accordingly, V-ATPase subunit mRNA levels are similar in human gliomas (grade II or IV) and in peritumoral tissues. A significant proportion (25%) of V-ATPase is present in the plasma membrane of both the C6 and the DI TNC1 astrocytic cells in culture. A bafilomycin-sensitive hyperpolarizing pump current through the plasma membrane was detected and measured after ionic channel inhibition, which corresponds most probably to an electrogenic transport of protons. This suggests that the plasma membrane V ATPase is active. It could contribute to cytoplasmic pH regulation in astrocytic cells. PMID- 11870876 TI - CNTF and CNTF receptor alpha are constitutively expressed by astrocytes in the mouse brain. AB - Ciliary neurotrophic factor (CNTF) is regarded as one of the signals that lead to astrocyte activation following central nervous system lesion. However, it is not clear if CNTF can directly initiate astrocytic responses to injury because CNTF levels are very low in most areas of the unlesioned brain and CNTF receptor (CNTFRalpha) expression by astrocytes has not yet been demonstrated in the intact brain. In the present study, the expression patterns of CNTF protein and CNTFRalpha mRNA were studied in the intact mouse brain using immunocytochemistry and in situ hybridization, respectively. These procedures were combined with immunocytochemistry for glial fibrillary acidic protein in order to identify CNTF and CNTFRalpha-expressing astrocytes. CNTF-immunoreactive astrocytes were exclusively found in white matter structures such as the optical tract, the corticospinal tract, and the fimbria-fornix. Gray matter astrocytes did not exhibit CNTF immunoreactivity. In contrast, CNTFRalpha mRNA-expressing astrocytes were found in gray matter areas, preferentially in the molecular layers of the cortex and hippocampus. White matter astrocytes did not show a detectable CNTFRalpha mRNA signal. These data demonstrate that both CNTF and its receptor are constitutively expressed by astrocytes in mouse brain. The nonoverlapping locations of astrocytes expressing detectable levels of CNTF and CNTFRalpha, respectively, may be related to distinct postlesional functions of these two glial cell populations. PMID- 11870877 TI - Stable transfection of cDNAs targeting specific steps of glycogen metabolism supports the existence of active gluconeogenesis in mouse cultured astrocytes. AB - In order to assess the participation of astrocytic gluconeogenesis in the synthesis of glycogen, mouse astrocytes were stably transfected with antisense cDNA of fructose-1,6-bisphosphatase (FBPase) and with sense and antisense cDNAs of glycogen synthase (GS). The antisenses of FBPase and GS have similar significant effect in decreasing astrocyte glycogen content by 60%, while sense GS significantly increased glycogen content by 100%. The FBPase activity was decreased by all three cDNAs used, while glycogen phosphorylase was not altered. The activity of GS was decreased by the antisense GS and increased by the sense GS. These data demonstrate that the gluconeogenesis in astrocytes is involved in the glycogenesis modulation. PMID- 11870878 TI - Retention of membrane-localized beta-catenin in cells lacking functional polycystin-1 and tuberin. AB - The tuberous sclerosis (TSC) 2 tumor suppressor gene encodes the protein tuberin, which has recently been shown to play a crucial role in the intracellular trafficking of polycystin-1, the product of the polycystic kidney disease (PDK) 1 gene. PKD1 is responsible for most cases of autosomal dominant polycystic kidney disease, which has been described as "neoplasia in disguise." Polycystin-1 is a membrane protein localized to adherens junctions in a complex containing E cadherin and alpha-, beta-, and gamma-catenins. To determine whether loss of membrane localization of polycystin-1 and E-cadherin affects the function of beta catenin, beta-catenin localization and signaling were characterized in tuberin null EKT2 and ERC15 cells and in tuberin-positive TRKE2 cells derived from polycystic, neoplastic, and normal rat kidney epithelial cells, respectively. EKT2 cells lacking tuberin because of inactivation of the Tsc2 gene fail to localize polycystin-1 and E-cadherin appropriately to these junctions. However, beta-catenin was retained at lateral cell membranes in both tuberin-null and tuberin-positive cells. Moreover, gene transcription mediated by beta-catenin T cell--specific transcription factor complexes showed no differences among EKT2, ERC15, and TRKE2 cells. Thus, beta-catenin was stably retained at the lateral cell membrane in tuberin-null renal cells lacking membrane-localized polycystin-1 and E-cadherin. These data suggest that, although loss of Tsc2 tumor suppressor gene function disrupts normal polycystin-1 function and membrane localization of E-cadherin, normal beta-catenin signaling is retained in tuberin-null cells. PMID- 11870879 TI - The chemopreventive compound curcumin is an efficient inhibitor of Epstein-Barr virus BZLF1 transcription in Raji DR-LUC cells. AB - To characterize the effects of inhibitors of Epstein-Barr virus (EBV) reactivation, we established Raji DR-LUC cells as a new test system. These cells contain the firefly luciferase (LUC) gene under the control of an immediate-early gene promoter (duplicated right region [DR]) of EBV on a self-replicating episome. Luciferase induction thus serves as an intrinsic marker indicative for EBV reactivation from latency. The tumor promoter 12-O-tetradecanoylphorbol-13 acetate (TPA) induced the viral key activator BamH fragment Z left frame 1 (BZLF1) protein ("ZEBRA") in this system, as demonstrated by induction of the BZLF1 protein-responsive DR promoter upstream of the luciferase gene. Conversely, both BZLF1 protein and luciferase induction were inhibited effectively by the chemopreventive agent curcumin. Semiquantitative reverse transcriptase (RT) polymerase chain reaction (PCR) further demonstrated that the EBV inducers TPA, sodium butyrate, and transforming growth factor-beta (TGF-beta) increased levels of the mRNA of BZLF1 mRNA at 12, 24, and 48 h after treatment in these cells. TPA treatment also induced luciferase mRNA with similar kinetics. Curcumin was found to be highly effective in decreasing TPA-, butyrate-, and TGF-beta-induced levels of BZLF1 mRNA, and of TPA-induced luciferase mRNA, indicating that three major pathways of EBV are inhibited by curcumin. Electrophoretic mobility shift assays (EMSA) showed that activator protein 1 (AP-1) binding to a cognate AP-1 sequence was detected at 6 h and could be blocked by curcumin. Protein binding to the complete BZLF1 promoter ZIII site (ZIIIA+ZIIIB) demonstrated several specific complexes that gave weak signals at 6 h and 12 h but strong signals at 24 h, all of which were reduced after application of curcumin. Autostimulation of BZLF1 mRNA induction through binding to the ZIII site at 24 h was confirmed by antibody induced supershift analysis. The present results confirm our previous finding that curcumin is an effective agent for inhibition of EBV reactivation in Raji DR CAT cells (carrying DR-dependent chloramphenicol acetyltransferase), and they show for the first time that curcumin inhibits EBV reactivation mainly through inhibition of BZLF1 gene transcription. PMID- 11870880 TI - Overexpression of a constitutively active form of c-src in skin epidermis increases sensitivity to tumor promotion by 12-O-tetradecanoylphorbol-13-acetate. AB - Transgenic mice were developed to study the role of c-src in epithelial tumorigenesis through targeted expression of a constitutively active form of murine c-src (src(529)). Src(529) was targeted to the interfollicular epidermis with the human keratin 1 (HK1) promoter. The skin phenotype of these mice was characterized by exaggerated epidermal hyperplasia and hyperkeratosis within the first week after birth. The severity of this phenotype correlated with overall src kinase activity, both of which subsided with age. Treatment of adult HK1.src(529) transgenic mice with the phorbol ester tumor promoter 12-O tetradecanoylphorbol-13-acetate resulted in an increase in epidermal hyperplasia and labeling index significantly greater than that seen in nontransgenic littermates. In addition, HK1.src(529) transgenic mice developed papillomas earlier and in significantly greater numbers compared with nontransgenic littermates in a standard initiation-promotion experiment. The data support the hypothesis that activation of c-src kinase plays a role in skin tumor promotion. PMID- 11870881 TI - Role of glutathione conjugate efflux in cellular protection against benzo[a]pyrene-7,8-diol-9,10-epoxide-induced DNA damage. AB - Glutathione (GSH) conjugation of (+)-anti-benzo[a]pyrene-7,8-diol-9,10-epoxide [(+)-anti-BPDE], the activated metabolite of benzo[a]pyrene, is believed to be an important mechanism in detoxification of this environmental and dietary carcinogen. Here, we demonstrate that the intracellular accumulation of GSH conjugate of (+)-anti-BPDE (BPD-SG) caused a statistically significant increase in (+)-anti-BPDE-induced DNA adduction. The relationship between intracellular accumulation of BPD-SG and (+)-anti-BPDE-induced DNA adduction was studied using a canine kidney epithelial cell line (MDCKII) and its variants overexpressing multidrug resistance transporter (MDR1) or canalicular multispecific organic anion transporter (cMOAT; also known as multidrug resistance protein 2). MDR1 and cMOAT are implicated in ATP-dependent efflux of anticancer drugs or GSH xenobiotic conjugates, or both. The GST activity toward (+)-anti-BPDE in parental MDCKII cells did not differ from that in subline overexpressing MDR1 (MDCKII MDR1) or cMOAT (MDCKII-cMOAT). Intracellular accumulation of BPD-SG, after a 5- or 10-min incubation with 1 microM (+)-anti-BPDE, was significantly higher in parental (41- to 67-fold) and MDCK II-MDR1 cells (31- to 43-fold) than in the MDCKII-cMOAT cells. Interestingly, the levels of DNA adducts of (+)-anti-BPDE, after a 30-min incubation with 0.1 or 0.5 microM [(3)H](+)-anti-BPDE, were significantly higher (about 2.1- and 1.7-fold, respectively) in parental cells than in the MDCKII-cMOAT cells. The results of the present study indicate that in addition to GSH conjugation, the efflux of BPD-SG may be essential for cellular protection against (+)-anti-BPDE-induced DNA damage. PMID- 11870882 TI - DNA methyltransferase and demethylase in human prostate cancer. AB - Recent studies have shown that cytosine-5 methylation at CpG islands in the regulatory sequence of a gene is one of the key mechanisms of inactivation. The enzymes responsible for CpG methylation are DNA methyltransferase (DNMT) 1, DNMT3a, and DNMT3b, and the enzyme responsible for demethylation is DNA demethylase (MBD2). Studies on methylation-demethylation enzymes are lacking in human prostate cancer. We hypothesize that MBD2 enzyme activity is repressed and that DNMT1 enzyme activity is elevated in human prostate cancer. To test this hypothesis, we analyzed enzyme activities, mRNA, and protein levels of MBD2 and DNMT1, DNMT3a, and DNMT3b in human prostate cancer cell lines and tissues. The enzyme activities of DNMTs and MBD2 were analyzed by biochemical assay. The mRNA expression was analyzed by reverse transcriptase-polymerase chain reaction and by Northern blotting. The protein expression was measured by immunohistochemistry with specific antibodies. The results of these experiments demonstrated that (1) the activity of DNMTs was twofold to threefold higher in cancer cell lines and cancer tissues, as compared with a benign prostate epithelium cell line (BPH-1) and benign prostatic hyperplasia (BPH) tissues; (2) MBD2 activity was lacking in prostate cancer cell lines but present in BPH-1 cells; (3) immunohistochemical analyses exhibited higher expression of DNMT1 in all prostate cancer cell lines and cancer tissues, as compared with BPH-1 cell lines and BPH tissues; (4) MBD2 protein expression was significantly higher in BPH-1 cells and lacking in prostate cancer cell lines and, in BPH tissues, MBD2 protein expression was poorly observed, as compared with no expression in prostate cancer tissues; and (5) mRNA expression for DNMT1 was upregulated in prostate cancer, as compared with BPH-1, and mRNA expression for MBD2 was found to be significantly expressed in all cases. The results of these studies clearly demonstrate that DNMT1 activity is upregulated, whereas MBD2 is repressed at the level of translation in human prostate cancer. These results may demonstrate molecular mechanisms of CpG hypermethylation of various genes in prostate cancer. PMID- 11870883 TI - Frequent microsatellite instability in lung cancer from chromate-exposed workers. AB - Although chromium has been the most extensively investigated metal with respect to mutagenicity and carcinogenicity, its genetic effects in humans are only partly understood. Our previous study demonstrated that lung cancer from chromate exposed workers infrequently (20%) displayed p53 gene mutations as well as a particular mutation pattern. In the present study, we examined the replication error (RER) and loss of heterozygosity (LOH) in 38 lung cancers from 28 chromate exposed workers (chromate lung cancer group) and in 26 lung cancer patients without chromate exposure (non-chromate lung cancer group), using six microsatellite markers containing CA repeats: D3S647 (3p23), D3S966 (3p21.3), D3S1289 (3p21.1), D5S346 (5q21-q22), D9S161 (9p21), and TP53 (17p13.1). The RER phenotype was defined as the presence of microsatellite instability (MSI) at two or more loci. Thirty (78.9%) of 38 tumors in the chromate lung cancer group exhibited RER. In contrast, only four (15.4%) of 26 tumors in the non-chromate lung cancer group exhibited RER. The frequency of RER in the chromate lung cancer group was significantly higher than that in the non-chromate lung cancer group (P < 0.0001). By contrast, the frequency of LOH at 3p, 5q, 9p, and 17p loci in tumors with chromate exposure was not significantly different from that in tumors without chromate exposure. In the chromate lung cancer group, the period of chromate exposure in workers with RER (24.5 +/- 6.7 yr) was significantly longer than that in workers without RER (17.0 +/- 3.5 yr) (P = 0.0046). In addition, a longer period of chromate exposure was associated with a tendency toward a higher frequency of MSI. This finding suggests that MSI may play a role in chromium induced carcinogenesis. In addition to our previous study of p53 mutations, the present findings suggest that the carcinogenic mechanism of chromate lung cancer may differ from that of non-chromate lung cancer. PMID- 11870884 TI - Restoration of wild-type conformation and activity of a temperature-sensitive mutant of p53 (p53(V272M)) by the cytoprotective aminothiol WR1065 in the esophageal cancer cell line TE-1. AB - The aminothiol WR1065, the active metabolite of the cytoprotector amifostine, exerts its antimutagenic effects through free-radical scavenging and other unknown mechanisms. In an earlier report, we showed that WR1065 activates wild type p53 in MCF-7 cells, leading to p53-dependent arrest in the G(1) phase of the cell cycle. To determine whether WR1065 activates p53 by modulating protein conformation, we analyzed its effects on p53 conformation and activity in the esophageal cancer cell line TE-1. This cell line contains a mutation in codon 272 of p53 (p53(V272M), with methionine instead of a valine), conferring temperature sensitive properties to the p53 protein. At the nonpermissive temperature (37 degrees C), p53(V272M) adopts the mutant p53 conformation (nonreactive with the antibody PAb1620), does not bind specifically to DNA, and is not activated in response to DNA-damaging treatment. However, treatment with 0.5-4 mM WR1065 partially restored wild-type conformation at 37 degrees C, stimulated DNA binding activity, and increased the expression of p53 target genes WAF-1, GADD45, and MDM2, leading to cell-cycle arrest in G(1). These results suggest that WR1065 activates p53 through a mechanism distinct from DNA-damage signaling, which involves modulation of p53 protein conformation. PMID- 11870886 TI - Direct analysis of highly oxidised organic aerosol constituents by on-line ion trap mass spectrometry in the negative-ion mode. AB - On-line ion trap mass spectrometry (ITMS) enables the characterisation of constituents of biogenic secondary organic aerosols in complex organic reaction mixtures. This real-time analysis is achieved by directly introducing the airborne particles into the ion source of the mass spectrometer. Negative-ion chemical ionisation at atmospheric pressure (APCI(-)) was used as the ionisation method of choice. The aerosols were generated from the gas-phase ozonolysis of two C10H16-terpenes (alpha-pinene and limonene), and investigated by performing on-line APCI(-)-ITMS(n). Highly oxidised compounds were tentatively identified as important particle-phase products. Based on recent investigations of low-energy collision-induced dissociation pathways of a wide range of deprotonated multifunctional carboxylic acid species derived from monoterpene precursors (Warscheid B, Hoffmann T. Rapid Commun. Mass Spectrom. 2001; 15: 2259), the formation of structurally different C10H16O5 and C10H16O6 species, such as acidic esters from alpha-pinene and aldo-hydroxycarboxylic acids from limonene, is proposed. PMID- 11870887 TI - High accuracy mass measurement of peptides with internal calibration using a dual electrospray ionization sprayer system for protein identification. AB - A dual-ESI-sprayer system was constructed and applied to achieve high accuracy of peptide mass measurement for protein identification by means of peptide mapping. Sample was introduced in one sprayer, and reference in the other, thus making internal calibration possible greatly enhancing the mass accuracy. Several samples were utilized to evaluate the reliability of this dual-ESI-sprayer system. The range of mass errors was 0.16-5.37 ppm. The peptide masses of tryptic digests of myoglobin (horse) were measured by the HPLC/dual-ESI-MS system, with mass deviations ranging from 0.01-7.67 ppm, and about 75% mass deviations below 5 ppm with 40% below 1[?]ppm. These peptide masses were utilized to perform database searching for protein identification, and compared to results obtained by external calibration. This comparison showed that the internal calibration provides a more reliable method of protein identification, with a much smaller number of required peptides for matching, and with less CPU time consumed for database searching. PMID- 11870890 TI - Novel phosphoryl derivatization method for peptide sequencing by electrospray ionization mass spectrometry. AB - A one-step phosphoryl derivatization method has been used in a peptide sequencing procedure for electrospray ionization tandem mass spectrometry (ESI-MS/MS). The sodiated derivatized peptides exhibit very simple dissociation patterns, in which two kinds of fragment ions, [b(n) + OH + Na]+ and [a(n) + Na]+, are formed. Since the amino acid residues are lost sequentially from the C-terminus, peptide sequences can be identified easily. The fragmentation efficiency of peptides increased as a result of the phosphorylation, and also provided peaks of useful intensity at lower m/z. A peptide with lysine at the C-terminus was derivatized and analyzed by ESI-MS/MS. Similar mass spectra, from which the sequence could be read out, were obtained. This is a novel derivatization method yielding neutral derivatives that should be suitable for peptide sequencing by LC/ESI-MS/MS. PMID- 11870891 TI - Effects of flow rate on high-throughput quantitative analysis of protein precipitated plasma using liquid chromatography/tandem mass spectrometry. AB - The effects of flow rate and column length on analyte response (peak area and height), total cycle time, column backpressure, and elution volume are presented. Rapid chromatographic separations and tandem mass spectrometric (MS/MS) detection are applied to the supernatant of protein-precipitated plasma standards containing four compounds from a drug discovery screen. The plasma samples were injected onto three C-18 columns (2 x 10,2.1 x 30 and 2.1 x 50 mm) at flow rates of 0.25, 0.50, 1.00 and 1.50 mL/min. The plasma samples were detected using a Sciex API 3000 tandem mass spectrometer operated in the Turbo Ionspray mode. A post-column split was used to maintain a flow rate of 0.25 mL/min into the mass spectrometer source to avoid differences in nebulization efficiency. The data show that diluted protein-precipitated plasma supernatants show average matrix effects (i.e. suppression) of 60.0% (2 x 10 mm), 89.3% (2 x 30 mm), and 76.7% (2 x 50 mm) of expected response at 10 ng/mL. Average matrix effects of 70.2% (2 x 10 mm), 88.9% (2 x 30 mm), and 81.2% (2 x 50 mm) of expected response at 1000 ng/mL plasma. The data also show if peak widths remain relatively constant, analytes are less sensitive as flow rates are increased. These data are consistent with the concentration-dependent relationship of ionspray in the range of flow rates studied. The data show that, while analyte response decreased proportionately to increases in flow rate, the analysis cycle times did not decrease proportionately. PMID- 11870892 TI - A straightforward means of coupling preparative high-performance liquid chromatography and mass spectrometry. AB - Flow splitting to a mass spectrometer is a common way of coupling a highly specific detector to preparative or semi-preparative high-performance liquid chromatography (HPLC) purification of combinatorial libraries, drug metabolites, and characterizable impurities. The sensitive mass spectrometer consumes only a small fraction of the analyte while providing online structure-specific detection, and its output can thus be used to trigger collection of the desired fraction. Coupling mass spectrometry to preparative HPLC is difficult due to the susceptibility of the detector to fouling under conditions of high analyte concentration or solute amount, or to changes in solvent composition. We report here on a device, the mass rate attenuator (MRA), which automatically produces split ratios over a range of 100:1 to 100 000:1 under programmable user control. The MRA is a flow-control device that periodically gates a small aliquot from one liquid stream into another. The design allows the user to set the frequency of the gating without interruption of the HPLC flow stream. The MRA also allows control of the volume of the aliquot that is transferred between the flow streams. This additional control, compared to passive splitting devices, facilitates optimization of the tubing connecting the separation, detection and collection events. We demonstrate that such optimization can reduce the volume of the collected fraction without compromising recovery, thus reducing the time spent in evaporating solvents to reclaim purified products. PMID- 11870894 TI - Multi-stage tandem mass spectrometry of metal cationized leucine enkephalin and leucine enkephalin amide. AB - We have examined the multi-stage collision induced dissociation (CID) of metal cationized leucine enkephalin, leucine enkephalin amide, and the N-acetylated versions of the peptides using ion trap mass spectrometry. In accord with earlier studies, the most prominent species observed during the multi-stage CID of alkali metal cationized leucine enkephalin are the [b(n) + 17 + Cat]+ ions. At higher CID stages (i.e. >MS(4)), however, dissociation of the [b2 + 17 + Cat]+ ion, a cationized dipeptide, results in the production of [a(n) -1 + Cat]+ species. The multi-stage CID of Ag+ cationized leucine enkephalin can be initiated with either the [b(n) -1 + Ag]+ or [b(n) + 17 + Ag]+ ions produced at the MS/MS stage. For the former, sequential CID stages cause, in general, the loss of CO, and then the loss of the imine of the C-terminal amino acid, to reveal the amino acid sequence. Similar to the alkali cationized species, CID of [b2 -1 + Ag]+ produces prominent [a(n) -1 + Ag]+ ions. The multi-stage CID of argentinated peptides is reminiscent of fragmentation observed for protonated peptides, in that a series of (b(n)) and (a(n)) type ions are generated in sequential CID stages. The Ag+ cation is similar to the alkali metals, however, in that the [b(n) + 17 + Ag]+ product is produced at the MS/MS and MS3 stages, and that sequential CID stages cause the elimination of amino acid residues primarily from the C-terminus. We found that N-acetylation of the peptide significantly influenced the fragmentation pathways observed, in particular by promoting the formation of more easily interpreted (in the context of unambiguous sequence determination) dissociation spectra from the [b2 + 17 + Li]+, [b2 + 17 + Na]+ and [b2 -1 + Ag]+ precursor ions. Our results suggest, therefore, that N-acetylation may improve the efficacy of multi-stage CID experiments for C-terminal peptide sequencing in the gas phase. For leucine enkephalin amide, only the multi-stage CID of the argentinated peptide allowed the complete amino acid sequence to be determined from the C-terminal side. PMID- 11870893 TI - Improved method for the simultaneous determination of d4T, 3TC and ddl intracellular phosphorylated anabolites in human peripheral-blood mononuclear cells using high-performance liquid chromatography/tandem mass spectrometry. AB - There is still a need for direct determination (i.e. without dephosphorylation) of nucleoside reverse transcriptase inhibitor (NRTI) triphosphorylated nucleotides in the peripheral-blood mononuclear cells (PBMCs) of HIV-positive patients. The objective of this paper was first to improve our previously described direct liquid chromatography/tandem mass spectrometry (LC/MS/MS) assay for stavudine triphosphate (d4T-TP). Preparation of PBMCs was modified to reduce degradation of d4T-TP during cell preparation and to simplify this step for routine use in clinical units. The performance of several HPLC columns was compared in order to improve the stability of peak shape over time. The SMT C(18) column was replaced by a Supelcogel ODP-50, thereby reducing two-fold the concentration of the first standard. Various internal standards were compared to optimize peak shape and remove an interfering peak in LC. 2-Chloroadenosine 5prime prime or minute-triphosphate was chosen as the most appropriate internal standard. Substitution of the narrowbore column by a microbore column (150 x 0.32 mm) is also presented and discussed. Secondly, this improved method was successfully applied to the simultaneous determination of d4T-TP, dideoxyadenosine triphosphate (ddA-TP) and lamivudine triphosphate (3TC-TP) in PBMCs, which is useful in view of the common use of NRTI combinations. The method was subsequently applied to clinical samples from HIV-positive patients receiving antiretroviral therapy containing d4T, ddl and/or 3TC. This method can be used simply and routinely on approximately 200 samples per week, using commercially available instruments and with a simple cell lysis as sample treatment. PMID- 11870896 TI - Detecting aristolochic acids in herbal remedies by liquid chromatography/serial mass spectrometry. AB - Targeted liquid chromatography/serial mass spectrometry (LC/MS/MS) analysis, using a quadrupole ion-trap mass spectrometer, permitted the detection of aristolochic acids I and II in crude 70% methanol extracts of multi-component herbal remedies without any clean-up or concentration stages. The best ionisation characteristics were obtained using atmospheric pressure chemical ionisation (APCI) and by including ammonium ions in the mobile phase. Limits of detection for aristolochic acids were influenced by the level of interference created by other components in the sample matrix. They were determined to be between 250 pg and 2.5 ng on-column within a matrix containing compounds extracted from 2 mg of herbal remedy. With a herbal remedy that only permitted the higher limit of detection, this sensitivity was sufficient to detect the aristolochic acids extracted from 0.1% dry weight of Aristolochia manshuriensis included in the preparation. PMID- 11870895 TI - Determination of rare earth elements in urine by electrothermal vaporization inductively coupled plasma mass spectrometry. AB - A method was developed for the determination of rare earth elements (REEs) in urine with electrothermal vaporization inductively coupled plasma mass spectrometry (ETV-ICPMS). The undiluted sample was directly injected into the graphite tube and trifluoromethane (Freon-23) was used as chemical modifier in order to reduce the vaporization temperature and the memory effect of most of the lanthanides. The detection limits were in the range 1-10 ng/L with relative standard deviation of 3-5% at concentration levels of 1microg/L, and less than 10 15% at 100 ng/L. Two different procedures, external calibration and a standard additions method, were evaluated to measure the concentration levels of lanthanides in the urine samples and the second procedure was considered to be the best choice for calibration in this work. The level of REEs in urine of 50 healthy volunteers was in the range 5-20 ng/L, above the detection limit of ETV ICPMS. PMID- 11870897 TI - Study of the neurotransmitter dopamine and the neurotoxin 6-hydroxydopamine by electrospray ionization coupled with tandem mass spectrometry. AB - Electrospray ionization combined with tandem mass spectrometry has been applied to a study of dopamine and 6-hydroxydopamine, an important neurotransmitter and a well-known neurotoxin, respectively. Both protonated and deprotonated molecules were observed for the two compounds. Upon collision-induced dissociation of protonated and deprotonated 6-hydroxydopamine molecules, the number of fragmentation pathways observed was greater than that observed with protonated and deprotonated dopamine molecules; the greater proclivity to fragment of the former is due to the 6-substituted hydroxyl group, which is para to the 3-OH group and ortho to the CH2CH2NH2 group. Furthermore, 6-hydroxydopamine showed a greater propensity to oxidize than did dopamine when sample solutions were kept uncovered in the air for 24 h prior to mass spectrometric examination. Radical structures of the four main oxidation products of 6-hydroxydopamine have been suggested on the basis of their product ion mass spectra; one or more of these oxidation products may be responsible for the cytotoxic property of 6 hydroxydopamine. PMID- 11870898 TI - Electrospray liquid chromatography/mass spectrometry fingerprinting of Acanthophis (death adder) venoms: taxonomic and toxinological implications. AB - Death adders (genus Acanthophis) are unique among elapid snakes in both morphology and venom composition. Despite this genus being among the most divergent of all elapids, the venom has been historically regarded as relatively quite simple. In this study, liquid chromatography/mass spectrometry (LC/MS) analysis has revealed a much greater diversity in venom composition, including the presence of molecules of novel molecular weights that may represent a new class of venom component. Furthermore, significant variation exists between species and populations, which allow for the LC/MS fingerprinting of each species. Mass profiling of Acanthophis venoms clearly demonstrates the effectiveness of this technique which underpins fundamental studies ranging from chemotaxonomy to drug design. PMID- 11870899 TI - Ion dispersion near parallel wire grids in orthogonal acceleration time-of-flight mass spectrometry: predicting the effect of the approach angle on resolution. AB - Ions experience small deflections in the vicinity of grids in accelerators and ion mirrors in time-of-flight (TOF) mass spectrometers. Recent experiments with an orthogonal acceleration (oa) TOF instrument have verified that the effect can significantly degrade resolution when ions approach grids at an angle deviating from 90 degrees. The phenomenon becomes significant only when ions have components of velocity at right angles to the wires of the grids. A model is presented in this study to predict this phenomenon for parallel wire grids. The fractional energy spread of ions (calculated in the static TOF-spectrometer frame of reference) scales directly with the approach angle of ions to the grid (as measured from normal approach). The energy spread also scales with the range of angles that is a consequence of the focusing effect in each gap between the wires of the grid. The equations imply that closely spaced parallel wire grids are best for deployment in oa-TOF systems where non-zero approach angles are unavoidable. Such grids are relatively impractical to manufacture and support but rectangular repeat cell grids with relatively few wires at right angles to the source axis are shown experimentally to introduce minimal energy spread. When these grids are rotated by 90 degrees, the resolution measured in a Q-TOF spectrometer is degraded in approximate agreement with the parallel wire model. A practical implication of this work is that grid transmissions in oa-TOF systems may be significantly increased without loss of resolution. Improvements of approximately 200% (V-mode) and approximately 400% (W-mode) in ion transmission were obtained in this study without compromising resolution. This was achieved with approximately 73% transmission grids and greater potential improvements in transmission are being realised since this study with approximately 89% transmission grids having similar geometry. PMID- 11870900 TI - Hydrogen/deuterium exchange for higher specificity of protein identification by peptide mass fingerprinting. AB - Genome sequencing projects produce large amounts of information that could be translated into potential protein sequences. Such amounts of material continuously increase protein database sizes. At present, 22 times more protein sequences are available in the SWISS-PROT and TrEMBL databases than 8 years ago in SWISS-PROT. One of the methods of choice for protein identification makes use of specific endoproteolytic cleavage followed by matrix-assisted laser desorption/ionisation mass spectrometric (MALDI-MS) analysis of the digested product. Since 1993, when this technique was first demonstrated, the conditions required for a correct identification have changed dramatically. Whilst 4-5 peptides with an uncertainty of 2-3 Da were sufficient for a correct identification in 1993, 10-13 peptides with less than 60 ppm mass error are now required for human and E. coli proteins. This evolution is directly related to the continuous increase in protein database sizes, which causes an increase in the number of false positive matches in identification results. Use of an information complement deduced from the primary protein sequence, in the process of identification by peptide mass fingerprints, can help to increase confidence in the identification results. In this article, we propose the exchange of labile hydrogen atoms with deuterium atoms to provide an alternative information complement. The exchange reaction with optimised techniques has shown an average 95% of hydrogen/deuterium (H/D) exchange on tryptic peptides. This level of exchange was sufficient to single out one or more peptides from a list of potential candidate proteins due to the dependence of H/D exchange on the peptide primary structure. This technique also has clear advantages in the identification of small proteins where direct protein identification is impaired by the limited number of endoproteolytic peptides. Then, information related to primary sequence obtained with this technique could help to identify proteins with high confidence without any expensive tandem mass spectrometry instruments. PMID- 11870901 TI - Detection and identification of quinonemethide triterpenes in Peritassa campestris by mass spectrometry. AB - Analysis of tingenone and tingenol quinonemethide triterpenes was made by gas chromatography/mass spectrometry (GC/MS) of their trimethylsilyl (TMS) ethers. An extra TMS group, in addition to those predicted from the known structures, is added to these compounds during the derivatization process. The electron impact mass spectra showed base peaks at m/z 549 and 623, respectively, for the TMS derivatives of tingenone and tingenol, and electrospray (ES) and collision activated dissociation (CAD) studies indicate that these ions correspond to losses of a methyl group from the derivatives studied. A mechanism, based on ES MS/MS studies, is suggested for the derivatization and fragmentation pattern. PMID- 11870902 TI - Electron ionization induced fragmentation of 4-hydroxychalcone derivatives. PMID- 11870903 TI - Tumor therapy. PMID- 11870904 TI - Evaluation of the anti-vascular effects of combretastatin in rodent tumours by dynamic contrast enhanced MRI. AB - The anti-vascular effects of the tubulin binding agent, disodium combretastatin A 4 3-O-phosphate (CA-4-P), have been investigated in the rat P22 carcinosarcoma by measurements of radiolabelled iodoantipyrine uptake and dynamic contrast-enhanced MRI. The iodoantipyrine estimates of absolute tumour blood flow showed a reduction from 0.35 to 0.04 ml g(-1) min(-1) 6 h after 10 mg kg(-1) CA-4-P and to <0.01 ml g(-1) min(-1) after 100 mg kg(-1). Tumour blood flow recovered to control values 24 h after 10 mg kg(-1) CA-4-P, but there was no recovery by 24 h after the higher dose. Dynamic contrast-enhanced MR images were obtained at 4.7 T, following injection of 0.1 mmol kg(-1) Gd-DTPA and analysed assuming a model arterial input function. A parameter, K(trans), which is related to blood flow rate and permeability of the tumour vasculature to Gd-DTPA, was calculated from the uptake data. K(trans) showed a reduction from 0.34 to 0.11 min(-1) 6 h after 10 mg kg(-1) CA-4-P and to 0.07 min(-1) after 100 mg kg(-1). Although the magnitude of changes in K(trans) was smaller than that in tumour blood flow, the time course and dose-dependency patterns were very similar. The apparent extravascular extracellular volume fraction, nu(e), showed a four-fold reduction 6 h after 100 mg kg(-1) CA-4-P, possibly associated with vascular shutdown within large regions of the tumour. These results suggest that K(trans) values for Gd DTPA uptake into tumours could be a useful non-invasive indicator of blood flow changes induced by anti-vascular agents such as combretastatin. PMID- 11870905 TI - Differential sensitivity of two adenocarcinoma xenografts to the anti-vascular drugs combretastatin A4 phosphate and 5,6-dimethylxanthenone-4-acetic acid, assessed using MRI and MRS. AB - The effects of two anti-vascular agents, combretastatin A4 phosphate (CA4P), and 5,6-dimethylxanthenone-4-acetic acid (DMXAA), on the perfusion of two human colon adenocarcinomas implanted in SCID mice, were assessed for up to 3 h using non invasive magnetic resonance imaging (MRI) and spectroscopy techniques (MRS). MRI measurements of GdDTPA inflow showed that treatment with CA4P had little effect on the perfusion of HT29 tumours. Localized (31)P MRS measurements also showed that the drug had no significant effect on tumour cell energy status, as assessed from the ratio of the integrals of the signals from inorganic phosphate (P(i)) and nucleoside triphosphates. However, after treatment with DMXAA, perfusion was reduced and the P(i)/NTP ratio increased, indicating that the HT29 tumour is susceptible to the action of this drug. The LS174T tumour model was susceptible to both CA4P and DMXAA, using the criteria of changes in GdDTPA inflow and P(i)/NTP ratio. PMID- 11870906 TI - Assessment of protamine-induced thrombosis of tumor vessels for cancer therapy using dynamic contrast-enhanced MRI. AB - Since the role of angiogenesis in cancer development has been recognized, the study of anti-angiogenic or anti-vascular therapeutic agents has become increasingly important for cancer treatment. Selective thrombosis is one approach towards this goal. Since many types of carcinoma accumulate large numbers of degranulating mast cells which will release heparin, intravenously injected protamine may bind to heparin, neutralize its anti-coagulant effect and induce thrombosis. In this work we studied the formation of thrombosis by using dynamic contrast enhanced MRI. The enhancement kinetics of the contrast medium measured before and after protamine treatment were compared to assess the thrombotic effect. The underlying concept was that if the vessels became clotted, the subsequently injected contrast medium could not be delivered into the tissue to cause enhancement. In addition to the tissue-specific changes, protamine may also induce systemic effect in the host. The therapy-induced changes measured in tumors were compared to changes in normal tissues: liver, kidney, and especially the muscle adjacent to tumor. The results showed that protamine induced pronounced changes in the tumor. However, the degree of change measured by MRI was not associated with the results of semiquantitative assessment of thrombosis assessed by histology, perhaps due to the heterogeneous nature of the tumor and the difficulty in sampling sufficient regions histologically. The protamine induced temporal effects were also studied. We demonstrated that protamine could induce selective thrombosis in tumors, and that the effect could last for several hours. Dynamic contrast-enhanced MRI can serve as a suitable means to investigate the mechanism of this novel approach to induce selective thrombosis for anti vascular cancer therapy. PMID- 11870907 TI - Combined vascular and extracellular pH imaging of solid tumors. AB - The unique physiological environment of solid tumors, frequently characterized by areas of poor flow, hypoxia, high lactate and low extracellular pH (pHe), influences vascularization, invasion and metastasis. Thus, vascularization and the physiological and metabolic environment play permissive (and conversely preventive) roles in invasion and metastasis. By using a multi-parametric approach of combined vascular and spectroscopic imaging, we can begin to evaluate which combinations of vascular, metabolic and physiological regions in a solid tumor represent the highest 'metastatic threat'. Here, we present measurements of pHe, vascular volume and permeability from co-localized regions within a solid tumor. These studies were performed for a group of metastatic (MDA-MB-231) and non-metastatic (MCF-7) human breast cancer xenografts. In this study, we have demonstrated the feasibility of such an approach, and presented methods of analyses to detect differences in patterns of combined parameters obtained from spatially co-registered regions in a solid tumor. PMID- 11870908 TI - MRI and fluorescence microscopy of the acute vascular response to VEGF165: vasodilation, hyper-permeability and lymphatic uptake, followed by rapid inactivation of the growth factor. AB - Vascular endothelial growth factor (VEGF) is one of the key growth factors regulating tumor angiogenesis and thus it is one of the primary targets for antiangiogenic therapy. The long-term effects of VEGF include induction of proliferation and migration of endothelial cells, tube formation and maintenance of the immature capillaries. The early effects of VEGF include vasodilation and increased permeability. We hypothesize that the early responses to VEGF can serve to develop a quantitative measure of the activity of VEGF, and therefore may be applicable for monitoring the efficacy of systemic suppression of VEGF signaling during antiangiogenic therapy. For that end we tested the ability of MRI and fluorescence microscopy to detect the early response to intradermal VEGF165 in nude mice. VEGF-induced local vasodilation and increased permeability was detected by intravenous administration of macromolecular biotin-BSA-GdDTPA(23) 30 min after intradermal administration of VEGF. Contrast leak showed saturation kinetics. Delayed contrast administration (90 min after intradermal administration of VEGF) resulted in low contrast leak and demonstrated that the saturation kinetics is not due to contrast equilibration between plasma and the interstitial space, but rather is due to suppression of vascular permeability. Permeability was restored by a second bolus of VEGF, showing that the saturation kinetics is primarily due to inactivation of the growth factor. Confocal microscopy of fluorescent BSA-FITC confirmed the permeability changes monitored by MRI. Moreover, confocal microscopy showed efficient lymphatic uptake of the extravasated contrast material specifically in regions of VEGF induced hyper permeability. PMID- 11870909 TI - Reproducibility of dynamic contrast-enhanced MRI in human muscle and tumours: comparison of quantitative and semi-quantitative analysis. AB - The purpose of this study was to determine the reproducibility of dynamic contrast-enhanced (DCE)-MRI and compare quantitative kinetic parameters with semi quantitative methods, and whole region-of-interest (ROI) with pixel analysis. Twenty-one patients with a range of tumour types underwent paired MRI examinations within a week, of which 16 pairs were evaluable. A proton density weighted image was obtained prior to a dynamic series of 30 T(1)-weighted spoiled gradient echo images every 11.9 s with an intravenous bolus of gadopentetate dimeglumine given after the third baseline data point. Identical ROIs around the whole tumour and in skeletal muscle were drawn by the same observer on each pair of examinations and used for the reproducibility analysis. Semi-quantitative parameters, gradient, enhancement and AUC (area under the curve) were derived from tissue enhancement curves. Quantitative parameters (K(trans), k(ep), v(e)) were obtained by the application of the Tofts' model. Analysis was performed on data averaged across the whole ROI and on the median value from individual pixels within the ROI. No parameter showed a significant change between examinations. For all parameters except K(trans), the variability was not dependent on the parameter value, so the absolute values for the size of changes needed for significance should be used for future reference rather than percentages. The size of change needed for significance in a group of 16 in tumours for K(trans), k(ep) and v(e) was -14 to +16%, -0.20 ml/ml/min (15%) and -1.9[?]ml/ml (6%), respectively (pixel analysis), and -16 to +19%, -0.23 ml/ml/min (16%) and +/- 1.9[?]ml/ml (6%) (whole ROI analysis). For a single tumour, changes greater than 45 to +83%, +/- 0.78 ml/ml/min (60%) and +/- 7.6 ml/ml (24%), respectively, would be significant (pixel analysis). For gradient, enhancement and AUC the size of change needed for significance in tumours was -0.24 (17%), -0.05 (6%) and -0.06 (8%), respectively for a group of 16 (pixel analysis), and +/- 0.96 (68%), +/- 0.20 (25%) and +/- 0.22 (32%) for individuals. In muscle, the size of change needed for significance in a group of 16 for K(trans), k(ep) and v(e) was -30 to +44%, +/- 0.81 ml/ml/min (61%) and +/- 1.7 ml/ml (13%). For gradient, enhancement and AUC it was +/- 0.09 (20%), +/- 0.02 (8%) and +/- 0.03 (12%). v(e), enhancement and AUC are highly reproducible DCE-MRI parameters. K(trans), k(ep) and gradient have greater variability, with larger changes in individuals required to be statistically significant, but are nevertheless sufficiently reproducible to detect changes greater than 14-17% in a cohort of 16 patients. Pixel analyses slightly improve reproducibility estimates and retain information about spatial heterogeneity. Reproducibility studies are recommended when treatment effects are being monitored. PMID- 11870910 TI - Reproducibility of quantitative dynamic MRI of normal human tissues. AB - The aim of the study was to establish the normal range and to evaluate the reproducibility of dynamic contrast enhanced MRI (DCE-MRI) parameter estimates in normal human pelvic tissues. Nineteen patients with prostate cancer, undergoing androgen deprivation treatment, had paired DCE-MRI examinations of the pelvis using spoiled gradient-echo sequences. Quantitative enhancement parameters were calculated for each examination: transfer constant (K(trans)), leakage space (v(e)) and maximum contrast medium accumulation (MCMA) of pelvic muscles, bone marrow and fat. Descriptive and reproducibility statistics were calculated: within-patient standard deviation (wSD), repeatability and within-patient coefficient of variation (wCV). The femoral head and ischiorectal fat showed large numbers of non-enhancing pixels (81 and 88%, respectively). The ischial bone marrow had the highest values of kinetic parameter estimates (K(trans) 0.554 min(-1), v(e) 18.5% and MCMA 0.164 mmol/kg). Muscle parameters values were lower (K(trans) 0.126-0.137 min(-1), v(e) 10.6-11.5% and MCMA 0.077-0.086 mmol/kg). The mean difference between paired examinations was not significantly different from zero for any parameter. v(e) and MCMA had the lowest wCV (between 19 and 29%). For individuals, a log(10) K(trans) change of approximately 0.90 in muscles and 0.52 in the ischium would be statistically significant. The corresponding absolute changes for v(e) are 6.7% in muscle and 13.6% in the ischium. For a group of 19 patients, small changes are statistically significant (muscle log(10) K(trans) 0.208 and v(e) 1.5% and ischium log(10) K(trans) 0.123 and v(e) 3.1%). Fat and the femoral head are unreliable tissues from which to obtain kinetic parameter estimates due to poor enhancement. v(e) and MCMA have smaller coefficient of variation than K(trans) in muscles and ischium. Reproducibility studies of normal and pathological tissues should be incorporated into clinical research protocols that measure treatment effects by DCE-MRI techniques. PMID- 11870911 TI - Assessing changes in tumour vascular function using dynamic contrast-enhanced magnetic resonance imaging. AB - Dynamic contrast-enhanced MRI (DCE-MRI) is widely used in the diagnosis and staging of cancer and is emerging as a promising method for monitoring tumour response to treatment. However, DCE-MR imaging techniques are still evolving and methods of image analysis remain variable and non-standard, and range from relative changes in the pattern of enhancement to pharmacokinetic modelling of contrast agent uptake. The combination of results from different institutions is therefore difficult and the sensitivities of different methods have not been compared. The purpose of this study is to investigate correlations between qualitative and quantitative methods of analysis for DCE-MR images from breast cancer patients undergoing neo-adjuvant chemotherapy. Fifteen patients underwent DCE-MRI examinations before and after one course of chemotherapy. Changes in the temporal pattern of signal enhancement, the rate and amplitude of enhancement and the volume transfer constant of contrast agent between the blood plasma and the extravascular extracellular space (EES), K(trans), and the EES fractional volume, nu(e), were determined. In addition, whole tumour region-of-interest analysis was compared with histogram analysis to investigate the extent of tumour heterogeneity. It was found that changes in the rate of enhancement correlated strongly with changes in K(trans) values (Kendall's tau = 0.68, P < 0.001). Furthermore, it was found that the shape of the signal enhancement curve only changed when the K(trans) values changed by 50% or more. Median K(trans) values determined following histogram analysis of pixel maps of K(trans) were approximately equal to those determined by whole tumour region-of-interest analysis. The absolute change in the K(trans) values correlated negatively with the pre-treatment values, particularly for responding patients. Thus, for higher pre-treatment K(trans) values, a greater decrease was observed. Greater changes were observed in the upper extremes of the K(trans) histogram than in the median values after one course of treatment. PMID- 11870912 TI - Breath-hold perfusion and permeability mapping of hepatic malignancies using magnetic resonance imaging and a first-pass leakage profile model. AB - We have applied a novel pharmacokinetic model of the distribution of contrast media to dynamic contrast-enhanced MRI data from patients with hepatic neoplasms. The model uses data collected during the passage of a bolus of contrast medium and allows breath-hold image acquisition. The aims of the study were to investigate the feasibility of permeability mapping using the first pass technique and breath-hold acquisitions, and to examine the reproducibility of the technique and the effect of the liver's dual vascular supply on the assumptions of the model. Imaging was performed in 14 patients with hepatic neoplasms. Dynamic data clearly demonstrated differences in the timing and shape of the contrast medium concentration-time course curve in the systemic arterial and portal venous systems. Mapping of the arrival time (T(0)) of contrast medium allowed identification of tissue supplied by the hepatic arteries and portal vein. Hepatic tumours all showed typical hepatic arterial enhancement. Repeated measurements of endothelial permeability surface area product (k(fp)) and relative blood volume (rBV), performed in five patients, showed excellent reproducibility with variance ratios (V(r)) of 0.134 and 0.113, respectively. Measurement of enhancing tumour volume was also highly reproducible (V(r) = 0.096) and this was further improved by the use of T(0) maps to identify pixels supplied by the hepatic artery (V(r) = 0.026). Estimates of k(fp) and rBV in normal hepatic tissue supplied by the portal vein were highly inaccurate and these pixels were identified by use of the T(0) parameter and excluded from the analysis. In conclusion, dynamic MRI contrast enhancement combined with a pharmacokinetic model of the distribution of contrast media in the first pass allows us to produce highly reproducible parametric maps of k(fp) and rBV from hepatic tumours that are supplied by the hepatic arterial system using breath hold acquisitions. PMID- 11870913 TI - Applications of sliding window reconstruction with cartesian sampling for dynamic contrast enhanced MRI. AB - Applications of dynamic contrast enhanced MR imaging are increasing and require both high spatial resolution and high temporal resolution. Perfusion studies using susceptibility contrast in particular require very high temporal resolution. The sliding window reconstruction is a technique for increasing temporal resolution. It has previously been applied to radial and spiral sampling, but these schemes require extensive correction and interpolation during image reconstruction. Fourier raw data can be reconstructed simply and quickly using the fast fourier transform (FFT). This paper presents a new Fourier-based sampling scheme and sliding window reconstruction that facilitates fast scanning without needing correction or interpolation. This technique can be used on virtually any MR scanner since it requires no specialized hardware. It is implemented here as a dual gradient echo sequence providing simultaneous T(1)- and T(2)*-weighted images with a time resolution of 1.1 s. PMID- 11870914 TI - Current awareness in NMR in biomedicine. PMID- 11870915 TI - The bioinformatics of molecular recognition. PMID- 11870916 TI - Elusive recognition determinants for ubiquitination. AB - How are proteins recognized as substrates for ubiquitination? Here we summarize insights from recent experiments that address this issue. These highlight the diversity and complexity of determinants for substrate recognition, and raise many questions for further investigation. PMID- 11870917 TI - Functional characterization of an anti-estradiol antibody by site-directed mutagenesis and molecular modelling: modulation of binding properties and prominent role of the V(L) domain in estradiol recognition. AB - The high-affinity monoclonal anti-estradiol antibody 9D3 presents a specificity defect towards estradiol-3-sulphate and 3-glucuronide conjugates incompatible with use in direct immunoassays. The corresponding single-chain variable fragment (scFv), cloned and produced in E. coli, exhibited a 10-fold lower affinity for estradiol (K(a)=1.2 x 10(9) M (-1)) and a slightly increased specificity defect for the 3-position. Site-directed mutagenesis revealed critical residues involved in estradiol recognition and produced mutants exhibiting up to a 3-fold increase of the binding affinity for estradiol and up to a 2-fold decrease of the cross reactivity with estradiol-3-sulphate. A comparative model of the antibody 9D3 estradiol complex was built in which the estradiol D-ring is buried into the binding pocket while the 3-, 6- and 7-positions are solvent exposed, agreeing with the lack of specificity for these three positions. Two potential alternative orientations of the A-ring, one close to CDR H3 and L2 loops, and the other one close to CDR H2 and L3 loops, have been considered for the docking of estradiol, none of which could be unambiguously privileged taking into account data from cross-reactivity measurements, photolabelling and mutagenesis studies. For both orientations, estradiol is stabilized by hydrogen bonding of the 17beta-OH group with TyrL36, His89 and GlnH35 in the first case, or TyrL36, only, in the second case and by van der Waals contacts from TyrL91 with alpha- or beta-face of estradiol, respectively, and from ValH95 and GlyH97 with the opposite face. To elucidate the molecular basis of antibody 9D3 specificity, as compared with that of another anti-estradiol antibody 15H11, single variable domains (V(H) and V(L)) and scFv hybrids have been constructed. The binding activity of V(L)9D3 as well as the specificity of the V(L)9D3/V(H)15H11 hybrid, both similar to antibody 9D3, revealed a prominent role of V(L) in estradiol recognition. These findings establish premises for antibody engineering to reduce cross-reactivity, especially with estradiol-3-conjugates. PMID- 11870918 TI - Distamycin A affects the stability of NF-kappaB p50-DNA complexes in a sequence dependent manner. AB - The effect of two different DNA minor groove binding molecules, Hoechst 33258 and distamycin A, on the binding kinetics of NF-kappaB p50 to three different specific DNA sequences was studied at various salt concentrations. Distamycin A was shown to significantly increase the dissociation rate constant of p50 from the sequences PRDII (5'-GGGAAATTCC-3') and Ig-kappa B (5'-GGGACTTTCC-3') but had a negligible effect on the dissociation from the palindromic target-kappaB binding site (5'-GGGAATTCCC-3'). By comparison, the effect of Hoechst 33258 on binding of p50 to each sequence was found to be minimal. The dissociation rates for the protein--DNA complexes increased at higher potassium chloride concentrations for the PRDII and Ig-kappaB binding motifs and this effect was magnified by distamycin A. In contrast, p50 bound to the palindromic target kappaB site with a much higher intrinsic affinity and exhibited a significantly reduced salt dependence of binding over the ionic strength range studied, retaining a K(D) of less than 10 pM at 150 mM KCl. Our results demonstrate that the DNA binding kinetics of p50 and their salt dependence is strongly sequence dependent and, in addition, that the binding of p50 to DNA can be influenced by the addition of minor groove-binding drugs in a sequence-dependent manner. PMID- 11870919 TI - Selective infection of E. coli as a function of a specific molecular interaction. AB - Selective infection of phage is when the bacterial infection depends on the specific molecular interaction between an antigen and a phage-displayed protein sequence such as an antibody. Engineering of the normal infection into pathways, directed by a specific protein--protein interaction, has raised several mechanistic questions. Here, we address the type of display and the affinity between the interacting pairs. The deleted phage R408d3 was used for the first time in selective infection and was shown to exhibit a superior performance compared to the VCSM13 phage. Furthermore, the affinity between the interacting pairs also affected the selective infection process and a correlation between affinity and infection efficiency was detected, thus implying that selective infection is the method of choice for selection of rare high-affinity interactions in molecular libraries. PMID- 11870920 TI - Coiled coil miniprotein randomization on phage leads to charge pattern mimicry of the receptor recognition determinant of interleukin 5. AB - Phage display was used to identify sequences that mimic structural determinants in interleukin5 (IL5) for IL5 receptor recognition. A coiled coil stem loop (CCSL) miniprotein scaffold library was constructed with its turn region randomized and panned for binding variants against human IL5 receptor alpha chain (IL5Ralpha). Competition enzyme-linked immunosorbent assays identified CCSL-phage selectants for which binding to IL5Ralpha was competed by IL5. The most frequently selected and IL5-competed CCSL-phage contain charged residues Arg and Glu in their turn sequences, in this regard resembling a beta strand sequence in the 'CD turn' region, of IL5, that has been proposed to present a key determinant for IL5 receptor alpha chain recognition. The most dominant CCSL-phage selectant sequence, PVEGRV, contains a negative/positive charge pattern similar to that seen in the original CD turn. To test the relatedness of CCSL-phage selectant sequences to the IL5 receptor recognition epitope, PVEGRV was grafted into the sequence 87--92 of a monomeric IL5. The resulting IL5 variant, [(87)PVEGRV(92)]GM1, was able to bind to IL5Ralpha in biosensor assays, to elicit TF-1 cell proliferation and to induce STAT5 phosphorylation in TF-1 cells. The results help discern sequence patterns in the IL5 CD turn region which are key in driving receptor recognition and demonstrate the utility of CCSL miniprotein scaffold phage display to identify local IL5 mimetic sequence arrangements that may ultimately lead to IL5 antagonists. PMID- 11870921 TI - Temperature differentially affects encounter and docking thermodynamics of antibody--antigen association. AB - Using BIACORE SPR, we have examined the mechanism of temperature effects on the binding kinetics of two closely related antibody Fabs (H10 and H26) which recognize coincident epitopes on hen egg-white lysozyme (HEL), and whose association and dissociation kinetics are best described by the two-step conformational change model which we interpret as molecular encounter and docking. Time-course series data obtained at a series of six temperatures (6, 10, 15, 25, 30 and 37 degrees C) showed that temperature differentially affects the rate constants of the encounter and docking steps. Docking is more temperature sensitive than the encounter step, and energetically less favorable at higher temperatures. At elevated temperatures, the time required for docking is longer and the apparent increase in off-rate reflects the greater proportion of the molecules failing to dock and remaining in the less stable encounter state. As a consequence, distribution of free energy change between the encounter and docking steps is altered. At physiological temperature (37 degrees C) the docking step of the H26 complex is energetically unfavorable and most complexes essentially do not dock. There is a significant decrease in total free energy change of the H26 complex at higher temperatures. Elevated temperature changes the rate-limiting step of H26--HEL association from the encounter to the docking step, but not that of H10--HEL. Our results indicate that the mechanism by which elevated temperature reduces the affinities of antigen--antibody complexes is to decrease the net docking rate, and/or stability of the docked complex; at higher temperatures, a smaller proportion of the complexes actually anneal to a more stable docked state. This mechanism may have broad applicability to other receptor--ligand complexes. PMID- 11870922 TI - Areas of brain activation in males and females during viewing of erotic film excerpts. AB - Various lines of evidence indicate that men generally experience greater sexual arousal (SA) to erotic stimuli than women. Yet, little is known regarding the neurobiological processes underlying such a gender difference. To investigate this issue, functional magnetic resonance imaging was used to compare the neural correlates of SA in 20 male and 20 female subjects. Brain activity was measured while male and female subjects were viewing erotic film excerpts. Results showed that the level of perceived SA was significantly higher in male than in female subjects. When compared to viewing emotionally neutral film excerpts, viewing erotic film excerpts was associated, for both genders, with bilateral blood oxygen level dependent (BOLD) signal increases in the anterior cingulate, medial prefrontal, orbitofrontal, insular, and occipitotemporal cortices, as well as in the amygdala and the ventral striatum. Only for the group of male subjects was there evidence of a significant activation of the thalamus and hypothalamus, a sexually dimorphic area of the brain known to play a pivotal role in physiological arousal and sexual behavior. When directly compared between genders, hypothalamic activation was found to be significantly greater in male subjects. Furthermore, for male subjects only, the magnitude of hypothalamic activation was positively correlated with reported levels of SA. These findings reveal the existence of similarities and dissimilarities in the way the brain of both genders responds to erotic stimuli. They further suggest that the greater SA generally experienced by men, when viewing erotica, may be related to the functional gender difference found here with respect to the hypothalamus. PMID- 11870923 TI - Simultaneous measurements of cerebral oxygenation changes during brain activation by near-infrared spectroscopy and functional magnetic resonance imaging in healthy young and elderly subjects. AB - Near infrared spectroscopy (NIRS) and functional magnetic resonance imaging (fMRI) both allow non-invasive monitoring of cerebral cortical oxygenation responses to various stimuli. To compare these methods in elderly subjects and to determine the effect of age on cortical oxygenation responses, we determined motor-task-related changes in deoxyhemoglobin concentration ([HHb]) over the left motor cortex in six healthy young subjects (age 35 +/- 9 years, mean +/- SD) and five healthy elderly subjects (age 73 +/- 3 years) by NIRS and blood-oxygen-level dependent (BOLD) fMRI simultaneously. The motor-task consisted of seven cycles of 20-sec periods of contralateral finger-tapping at a rate as fast as possible alternated with 40-sec periods of rest. Time-locked averages over the seven cycles were used for further analysis. Task-related decreases in [HHb] over the motor cortex were measured by NIRS, with maximum changes of -0.83 +/- 0.38 mumol/L (P < 0.01) for the young and -0.32 +/- 0.17 mumol/L (P < 0.05) for the elderly subjects. The BOLD-fMRI signal increased over the cortex volume under investigation with NIRS, with maximum changes of 2.11 +/- 0.72% (P < 0.01) for the young and 1.75 +/- 0.71% (P < 0.01) for the elderly subjects. NIRS and BOLD fMRI measurements showed good correlation in the young (r = -0.70, r(2) = 0.48, P < 0.001) and elderly subjects (r = -0.82, r(2) = 0.67, P < 0.001). Additionally, NIRS measurements demonstrated age-dependent decreases in task-related cerebral oxygenation responses (P < 0.05), whereas fMRI measurements demonstrated smaller areas of cortical activation in the elderly subjects (P < 0.05). These findings demonstrate that NIRS and fMRI similarly assess cortical oxygenation changes in young subjects and also in elderly subjects. In addition, cortical oxygenation responses to brain activation alter with aging. PMID- 11870924 TI - Exact multivariate tests for brain imaging data. AB - In positron emission tomography (PET) and functional magnetic resonance imaging (fMRI) data sets, the number of variables is larger than the number of observations. This fact makes application of multivariate linear model analysis difficult, except if a reduction of the data matrix dimension is performed prior to the analysis. The reduced data set, however, will in general not be normally distributed and therefore, the usual multivariate tests will not be necessarily applicable. This problem has not been adequately discussed in the literature concerning multivariate linear analysis of brain imaging data. No theoretical foundation has been given to support that the null distributions of the tests are as claimed. Our study addresses this issue by introducing a method of constructing test statistics that follow the same distributions as when the data matrix is normally distributed. The method is based on the invariance of certain tests over a large class of distributions of the data matrix. This implies that the method is very general and can be applied for different reductions of the data matrix. As an illustration we apply a test statistic constructed by the method now presented to test a multivariate hypothesis on a PET data set. The test rejects the null hypothesis of no significant differences in measured brain activity between two conditions. The effect responsible for the rejection of the hypothesis is characterized using canonical variate analysis (CVA) and compared with the result obtained by using univariate regression analysis for each voxel and statistical inference based on size of activations. The results obtained from CVA and the univariate method are similar. PMID- 11870925 TI - Anatomically informed basis functions in multisubject studies. AB - We describe the use of anatomically informed basis functions (AIBF) in the analysis of multisubject functional imaging studies. AIBF are used to specify an anatomically informed spatial model that embodies anatomical knowledge for the statistical analysis of neuroimaging data. In a previous communication, we showed how AIBF can be used to incorporate prior anatomical constraints in single subject functional magnetic resonance image (fMRI) analyses to augment their anatomical precision. In this paper, we extend AIBF such that it can be applied to multisubject studies using fMRI or PET. The key concept is that, after spatial normalization, a canonical cortical surface can be used to generate a forward model of signal sources for all subjects. By estimating the hemodynamic signal in this canonical AIBF-space and then projecting it back into the voxel-space, one effectively extracts functional activity that is smooth, within and only within, the cortical sheet while attenuating other components unrelated to the physiological process of interest. The ensuing procedure can be considered as a highly non-stationary, anisotropic anatomically informed [de]convolution or smoothing. It is shown that this procedure offers various advantages compared to existing conventional methods for the analysis of multisubject studies, in particular it is more sensitive to underlying activations. PMID- 11870926 TI - Monte Carlo simulation studies of EEG and MEG localization accuracy. AB - Both electroencephalography (EEG) and magnetoencephalography (MEG) are currently used to localize brain activity. The accuracy of source localization depends on numerous factors, including the specific inverse approach and source model, fundamental differences in EEG and MEG data, and the accuracy of the volume conductor model of the head (i.e., the forward model). Using Monte Carlo simulations, this study removes the effect of forward model errors and theoretically compares the use of EEG alone, MEG alone, and combined EEG/MEG data sets for source localization. Here, we use a linear estimation inverse approach with a distributed source model and a realistic forward head model. We evaluated its accuracy using the crosstalk and point spread metrics. The crosstalk metric for a specified location on the cortex describes the amount of activity incorrectly localized onto that location from other locations. The point spread metric provides the complementary measure: for that same location, the point spread describes the mis-localization of activity from that specified location to other locations in the brain. We also propose and examine the utility of a "noise sensitivity normalized" inverse operator. Given our particular forward and inverse models, our results show that 1) surprisingly, EEG localization is more accurate than MEG localization for the same number of sensors averaged over many source locations and orientations; 2) as expected, combining EEG with MEG produces the best accuracy for the same total number of sensors; 3) the noise sensitivity normalized inverse operator improves the spatial resolution relative to the standard linear estimation operator; and 4) use of an a priori fMRI constraint universally reduces both crosstalk and point spread. PMID- 11870927 TI - Carotid and cerebral angiography performed by cardiologists: cerebrovascular complications. AB - The management of extracranial carotid artery disease is primarily concerned with the prevention of acute stroke. In order to understand the current risks of carotid angiography performed by interventional cardiologists, we undertook a retrospective study to determine the neurologic complications in patients who underwent selective cerebral angiography. All patients undergoing studies that were limited to diagnostic aortic arch angiography and selective four-vessel cerebral angiography in the cardiac catheterization laboratories during the past 6 years were included in this study. Hospital records were reviewed to determine any in-hospital cerebrovascular complications following carotid angiography, ranging from transient ischemic attack to major disabling stroke or death. A total of 189 consecutive patients underwent 191 diagnostic studies limited to aortic arch and four-vessel cerebral angiography in the cardiac catheterization laboratories between 1 January 1995 and 31 December 2000. Only one (0.52%) neurological complication, a minor stroke, occurred in our study population. There were no transient ischemic attacks, major strokes, or death. We have shown that experienced interventional cardiologists can perform diagnostic aortic arch and selective carotid and vertebral angiography in a cardiac catheterization laboratory with a very low complication rate. Because the risks of angiography add to those of revascularization of the carotid artery, the most highly skilled angiographer, regardless of primary specialty, should perform these studies. PMID- 11870928 TI - Percutaneous repair of abdominal aortic aneurysms using the AneuRx stent graft and the percutaneous vascular surgery device. AB - Endovascular exclusion of abdominal aortic aneurysms (AAAs) was developed in an effort to treat patients who were at high risk for complications following standard surgical repair. Stent grafts used for endovascular repair of AAAs require the use of large-bore sheaths and surgical exposure of the common femoral arteries (CFAs). To decrease the invasiveness of AAA repair, we attempted to perform the procedure percutaneously utilizing the Prostar XL Percutaneous Vascular Surgery Device and the preclose technique. Thirty patients underwent an attempted percutaneous AAA repair. These patients were followed prospectively to assess the success of the procedure. Twenty-eight patients (93%) had successful percutaneous repair of both CFA access sites. One patient had inadequate hemostasis of the 22 Fr CFA entry site and one patient had inadequate hemostasis of the 16 Fr CFA entry site. Both of these CFA sites underwent open surgical repair. The rate of successful repair of the 22 Fr CFA access site was 29 of 30 (96%); for the 16 Fr CFA access site, 29 of 30 (96%). No in-hospital groin complications were seen. The procedure time was 105 +/- 21 min. The estimated blood loss was 90.6 +/- 50 cc. The hemoglobin loss was 1.54 +/- 0.89 mg/dL and the hematocrit loss was 5.04% +/- 2.8%. Complete percutaneous endoluminal AAA repair is feasible using the preclose technique. CFAs with sheaths up to 22 Fr can be safely and successfully accessed and repaired percutaneously using this technique. This method provides secure hemostasis and reduces the invasiveness of procedures requiring large-bore sheaths. PMID- 11870929 TI - Percutaneous femoral arterial closure in AAA endograft: advantages and disadvantages. PMID- 11870930 TI - Percutaneous coronary intervention with bivalirudin anticoagulation, immediate sheath removal, and early ambulation: a feasibility study with implications for day-stay procedures. AB - We assessed the feasibility and safety of a strategy of transfemoral percutaneous coronary intervention (PCI) with bivalirudin anticoagulation, immediate sheath removal, early ambulation, and, where possible, same-day discharge in 100 consecutive patients. Ambulation was achieved by 2 hr 30 min in 85% of patients and same-day discharge in 26%. PCI was angiographically successful in 97%. In hospital, there were no deaths or Q-wave myocardial infarctions. One patient suffered a non-Q-wave infarction, another in-hospital surgical revascularization and one required blood transfusion for rectal bleeding. Femoral access site hematoma > 5 cm diameter occurred in two patients. In addition, by 1 month there had been one death (at 10 days) and one pseudoaneurysm treated nonsurgically. In this preliminary study, the strategy of bivalirudin bolus anticoagulation, immediate sheath removal, and 2-hr ambulation after PCI appeared safe, with same day discharge possible in 26% of unselected patients with stable or unstable angina. PMID- 11870931 TI - Unrestricted availability of intracoronary stents is associated with decreased abrupt vascular closure rates and improved early clinical outcomes. AB - The purpose of this study was to determine whether the unrestricted availability of intracoronary stents is affecting abrupt vascular closure rates and early clinical outcomes. Intracoronary stents have improved procedural outcome despite their application in more complex lesions, but the impact of unrestricted availability of stents on abrupt closure and early clinical outcomes is unknown. Two cohorts were identified retrospectively from a coronary intervention registry: patients treated between 1988 and 1992 (n = 3,617) when stents were not generally available, and patients treated between 1994 and 1997 (n = 4,518) when stents were freely available. The late cohort patients consisted of more females; it also had greater comorbidity and more complex presentation. However, there was a significant reduction in the abrupt closure rates between the two time periods (7% vs. 4%, P < 0.0001). After adjusting for female gender, thrombus, and lesion angulation, the late cohort remained significantly associated with decreased odds of abrupt closure (odds ratio [OR] = 0.62, 95% upper and lower confidence intervals [CI] = 0.52-0.75, P = 0.0001). The late cohort also had decreased in hospital mortality (OR = 0.91, CI = 0.86-0.95, P = 0.0002) and major adverse clinical event (OR = 0.93, CI = 0.91-0.96, P = 0.0001). Unrestricted availability of intracoronary stents is associated with reduced abrupt closure rates and improved early clinical outcomes among patients undergoing coronary intervention despite treatment of high-risk patients. PMID- 11870932 TI - Stenting very small coronary narrowings (< 2 mm) using the biocompatible phosphorylcholine-coated coronary stent. AB - Published data regarding stenting very small arteries are still limited. The BiodivYsio stent is a new stent coated with phosphorylcholine, a biocompatible molecule designed to reduce the formation of thrombus and potentially the risk of restenosis. The feasibility, safety, and efficacy of implantation of the 2.0 mm coated coronary stent were prospectively studied. We studied 97 patients from three centers who underwent elective, urgent, or bailout implantation of 106 BiodivYsio mini-stents (2.0 mm) in 101 lesions. Forty percent of lesions had unfavorable characteristics (type B2 or C) and 16% had thrombus and/or chronic total occlusion. Successful stent deployment was achieved in 100/101 lesions (99%). MLD increased from 0.49 +/- 0.31 mm to 1.89 +/- 0.41 mm and diameter stenosis decreased from 89% +/- 7% to 5.6% +/- 6%. Small vessel stenting was the only procedure in 71% patients. There was one acute stent thrombosis case. During 6-month follow-up, none died, one had MI, and one was referred to CABG due to nontarget lesion progression. Angiographic restenosis that required target lesion revascularization was performed in 8/18 that had catheterization due to chest pain or significant ischemia. Most patients improved in their clinical symptoms. The rate of major adverse cardiac events was 4.1% at 30-day and 10.3% at 6-month follow-up. This initial clinical experience indicates that the implantation of 2.0 mm stents coated with phosphorylcholine appears to be safe and efficacious in the treatment of complex coronary lesions and is associated with low target vessel revascularization rate in spite of the very small vessel diameter. PMID- 11870933 TI - Stenting vs. balloon angioplasty with provisional stenting for the treatment of vessels with small reference diameter. AB - A consecutive series of interventions in vessels with reference diameter < or = 2.75 mm was retrospectively analyzed according to preprocedure strategy: balloon angioplasty with provisional stenting (PTCA group, 73 patients) and primary stenting (PS group, 122 patients). In the PS group, there were more patients with single-vessel disease (54.1% vs. 37.0%; P = 0.021), less patients with three vessel disease (9.0% vs. 24.7%; P = 0.003), more LAD interventions (54.9% vs. 31.5; P = 0.002), and less left circumflex interventions (22.1% vs. 45.2%; P < 0.001). Reference diameter was larger in the PS group (2.28 +/- 0.35 mm vs. 2.11 +/- 0.36 mm; P = 0.001). Provisional stenting was performed in 39.7% of PTCA group. At long-term outcome, the incidence of composite major events was similar between the PTCA and the PS groups (20.5% vs. 17.2%, respectively; P = NS). Treatment of small vessels with balloon dilatation and provisional stenting or with primary stenting yielded similar late outcomes. Operators' choice of treatment strategy was based on particular angiographic characteristics. PMID- 11870934 TI - Heparin and coumadin versus acetylsalicylic acid for prevention of restenosis after coronary angioplasty. AB - The purpose of the present study was to determine whether postprocedural antithrombotic therapy with prolonged heparin infusion followed by 6 months of oral anticoagulation in addition to acetylsalicylic acid (ASA) reduces the incidence of angiographic restenosis after successful PTCA. One hundred ninety one patients with uncomplicated PTCA were randomized into two groups: one group was discharged with ASA 100 mg only (G1) and the other group was additionally treated with 12-24 hr of heparin infusion and overlapping oral anticoagulation with coumadin for 6 months (G2). The two groups were comparable with respect to age, gender, coronary risk profile, clinical presentation, and angiographic lesion characteristics. Stents were implanted in 33% and 36% of the G1 and G2 patients, respectively. In-hospital myocardial infarction occurred in 4% of the G1 and 3% of the G2 patients. One patient in G1 died of subacute stent thrombosis (day 3). Six-month angiographic follow-up was obtained in 90% of G1 patients and 94% of G2 patients. Restenosis occurred in 30% and 33% of the patients and mean diameter stenoses at follow-up were 40% +/- 28% and 39% +/- 24%, respectively. Thrombin inhibition with heparin infusion followed by 6 months of oral anticoagulation did not reduce angiographic restenosis among patients undergoing PTCA with or without stent implantation. The occurrence of acute ischemic complications was also comparable in the two groups. PMID- 11870935 TI - Diabetic patients treated with abciximab and intracoronary stenting. AB - Diabetic patients are at greater risk for restenosis, recurrent ischemia, and complications following angioplasty than are their nondiabetic counterparts. This is a retrospective study identifying diabetic patients who were treated with abciximab and intracoronary stenting during the period of January 1997 to December 1999. Abciximab was administered to 268 of 707 diabetic patients who received intracoronary stents from 1997 to 1999. The abciximab group contained a higher number of patients with severe ventricular dysfunction and high-grade lesions. Primary endpoints of all-cause mortality, same-vessel revascularization, CABG, TVR, and postprocedural myocardial infarction were similar for both groups. The abciximab group had reduced rates of readmission for cardiac reasons during all follow-up periods. The trends toward improvement of mortality, surgical or percutaneous revascularization, and cardiac readmissions suggest the effect of abciximab may provide benefit for up to 9 months for higher-risk diabetic patients. PMID- 11870936 TI - Polytetrafluoroethylene-covered stent and coronary artery aneurysms. AB - Angiographically detected coronary aneurysms (i.e., coronary segment greater then 1.5 times the normal artery) have an incidence of 0.3%-4.9% among patients undergoing coronary angiography and have been reported after an intervention procedure with a frequency of 2%-10%. The indication for treatment and the best modality still need to be defined. Some authors reported the successful treatment of coronary aneurysms with the polytetrafluoroethylene (PTFE)-covered stent implantation, supporting the role of this strategy. In our institution, from September 1997 to December 1999 eight PTFE-covered stents were implanted to treat seven coronary aneurysms in seven patients. All aneurysms were successfully treated by the PTFE-covered stent. In one case, there was the necessity of an additional PTFE stent to cover the aneurysm completely. In no case did the loss of stent occur. No in-hospital MACE occurred. At 35 +/- 8 (21-44) months, six patients were symptom-free. Angiographic follow-up was performed in all patients at 10 +/- 6 months. Restenosis occurred in one patient (14%) who had repeat percutaneous coronary interventions. This preliminary experience suggests that PTFE-covered stent may be useful in the treatment of coronary artery aneurysms. PMID- 11870937 TI - Clinical and angiographic determinants of adverse cardiac events in patients with stent restenosis. AB - Patients with angiographically proven stent restenoses do not necessarily develop adverse cardiac events. Which clinical, procedural, or angiographic parameters relate to the development of adverse cardiac events among these patients has not been determined. This study included 155 patients (167 stented lesions) with angiographically proven restenosis (> or = 50% diameter stenosis) within the stent or at its margins in routine follow-up angiograms that was obtained at 6.5 +/- 3.6 months after successful stenting. Thirty-six patients (22%) had adverse cardiac events (including unstable angina necessitating target lesion revascularization, acute myocardial infarction, or cardiac death) during follow up and 119 patients (78%) were event-free. These two groups of patients were compared to determine the parameters related to adverse cardiac events. Univariate determinants of adverse events included hypertension (P = 0.023), unstable angina at initial presentation (P = 0.002), target lesion in proximal left anterior descending artery (P = 0.041), TIMI grade 0-2 flow in follow-up angiograms (p < 0.001), impaired left ventricular function at follow-up (P = 0.002), follow-up minimal lumen diameter < or = 0.6 mm (P = 0.003), follow-up diameter stenosis > 75% (P = 0.005), late loss > 2 mm (P = 0.01), and loss index > 1.127 (P < 0.001). Multivariate analysis demonstrated hypertension (odds ratio, OR, = 3.6; P = 0.019), unstable angina at initial presentation (OR = 2.6; P = 0.007), TIMI grade 0-2 flow at follow-up (OR = 2.8; P = 0.05), impaired LV function at follow-up (OR = 4.2; P = 0.004), and loss index > 1.127 (OR = 3.6; P = 0.017) as independent risk factors for adverse cardiac events. Classification and regression tree analysis identified loss index > 1.127 and impaired LV function as the two strongest determinant of adverse cardiac event. Therefore, hypertensive patients whose initial clinical presentation were unstable angina should be managed carefully to optimize the angiographic results and, most importantly, followed up more closely for development of impaired LV function after coronary stenting in order to prevent the occurrence of adverse cardiac event at follow-up. PMID- 11870938 TI - Post-PCI cardiac events? The answer is the lumen, or is it in the wall? PMID- 11870939 TI - Mini-invasive strategy in acute coronary syndromes: direct coronary stenting using 5 Fr guiding catheters and transradial approach. AB - The purpose of this study was to assess the feasibility and safety of direct coronary stenting in acute coronary syndromes using 5 Fr guiding catheters by transradial approach. A series of 119 patients with an acute coronary syndrome (unstable angina, n = 55; acute myocardial infarction, n = 45; recent acute myocardial infarction, n = 19) explored by transradial approach and eligible for direct stenting were included. A large proportion of patients (52%) was treated during the procedure by platelet IIb/IIIa receptor blockade. Only Medtronic 5 Fr guiding catheters were used in this study. Direct coronary stenting was attempted in all 119 highly selected patients. Failure of direct stenting was observed in only five cases (3.9%) and the stent successfully retrieved in each case in the 5 Fr guiding catheter. In these five cases, balloon predilation was performed and then the stent implanted successfully. Different stents were used: ACS stent (54%), AVE stent (33%), Velocity stent (10%), Nir stent (3%), with diameter ranging from 2.5 to 4 mm. In four cases, the dilation was finally performed using 6 Fr guiding catheters because the backup of the 5 Fr catheter was considered to be too low (3%). No vascular access site complications occurred in this series of patients. We conclude that direct coronary stenting using transradial approach and 5 Fr guiding catheters yields excellent procedural success rate. In the setting of acute coronary syndromes requiring platelet IIb/IIIa receptor blockade or after failure of thrombolysis, this mini-invasive strategy is very attractive because of the low risk of access site complications. PMID- 11870940 TI - Four-dimensional analysis of cyclic changes in coronary artery shape. AB - The objective of this study was to derive a method for quantifying the dynamic geometry of coronary arteries. Coronary artery geometry plays an important role in atherosclerosis. Coronary artery geometry also influences the performance of coronary interventions. Conversely, implantation of stents may alter coronary artery geometry. Clinical tools to define vessel shape have not been readily available. Using a Frenet-Serret curvature analysis applied to 3D reconstruction data derived from standard coronary angiograms, 21 coronary arteries were analyzed at end-diastole (ED) and end-systole (ES). Vessels were divided anatomically: type 1 consisted of vessels lying in the AV groove (left circumflex, right coronary) and type 2 consisted of vessels overlying actively contracting myocardium (left anterior descending, diagonal, obtuse marginal, right ventricular marginal, posterior descending, posterolateral). Vessel segments were analyzed by assessing the changes in curvature, torsion, and discrete flexion points (FPs), areas of systolic bending in the arterial contour. The curvature from ED to ES of type 1 vessels was unchanged (-0.02 +/- 0.03 cm( 1)), while the curvature change of type 2 vessels showed a 38% increase (0.33 +/- 0.04 cm(-1); P < 0.001). Type 1 vessels had fewer FPs per vessel than type 2 vessels (0.38 +/- 0.18 and 2.40 +/- 0.23 FP/vessel, respectively; P < 0.001). FPs were more common in distal segments and branch vessels. A method to quantify cyclic changes in coronary artery shape was applied to 3D data sets derived from standard coronary angiograms. Coronary arteries undergo a cyclic change in shape resulting in changes in overall curvature as well as formation of discrete flexion points. These changes in vessel shape are asymmetrically distributed in coronary arteries. PMID- 11870941 TI - Catheter therapy of Swiss cheese ventricular septal defects using the Amplatzer muscular VSD occluder. AB - The medical and surgical management of patients with multiple muscular ventricular septal defects (VSDs) is associated with morbidity and mortality. Three children with Swiss cheese VSDs were treated with transcatheter occlusion of their multiple defects using the Amplatzer muscular VSD occluder. Seventeen defects were closed in five catheterization procedures. One patient had three devices placed in two procedures, the second had five defects closed in one procedure, and the third had nine defects closed in two procedures. Two patients had previously been treated with pulmonary artery banding and required subsequent surgical band removal. There was immediate reduction in the left-to-right shunting and clinical improvement in all patients. Complications included the need for blood transfusion during the two longest procedures and tricuspid valve regurgitation in one. Transcatheter occlusion of multiple VSDs is an acceptable alternative or adjunct to surgical therapy for these complex patients. PMID- 11870942 TI - Transradial coronary brachytherapy with the Novoste Beta-Rail system. AB - We report our initial experience in 10 consecutive patients who underwent transradial coronary brachytherapy for in-stent restenosis using a 90Sr/Y source and the Novoste Beta-Rail system. In all patients, procedures were successfully completed using a right transradial approach. We performed the procedures with the Beta-Rail catheter using 7 Fr (Zuma II, Medtronic, MN; n = 5) or 8 Fr (Cordis, Miami, FL; n = 5) guiding catheters. All lesions were successfully dilated and no additional stent was inserted. We used a 40 mm source (n = 3) or a 60 mm source (n = 7) with manual stepping in four cases. In three cases, we did one stepping, and in one case, we did three steppings. The mean dwell time was 195 plus minus 44 sec. The mean delivered dose was 23 +/- 3 Gy at 2 mm distance from the source. No radiation treatment was interrupted. Mean fluoroscopy time was 26 +/- 13 min. Procedural success was achieved in all patients. Three patients had mild CK elevations (< 3 times upper normal limit). All patients were pretreated with clopidogrel (300 mg) and combined treatment with aspirin + clopidogrel is to be continued for at least 1 year. Clinical follow-up up to 3 months has not yielded any complication and all patients have remained free from angina. PMID- 11870943 TI - Simplified intracardiac electrocardiography for Ebstein's anomaly. PMID- 11870944 TI - An unusual case of left internal mammary artery ostial disease: clarifying role of intravascular ultrasound. AB - Stenosis at the origin of the left internal mammary artery graft is rare. We present a case with a suspected stenosis involving the origin of the left internal mammary artery that conventional angiography failed to demonstrate convincingly. Intravascular ultrasound illustrated a severe stenosis and the patient underwent successful stenting of the left internal mammary artery origin. The intravascular ultrasound finding of a dissection flap, just distal to the left internal mammary artery origin, suggests that local trauma to the vessel from prior catheterization procedures may have been responsible for the progressive narrowing at the left internal mammary artery ostium. PMID- 11870945 TI - First use of intracoronary beta-radiation to prevent recurrent in-stent restenosis in a transplanted heart. PMID- 11870946 TI - Successful balloon valvuloplasty in an adult patient with severe pulmonic stenosis and aneurysmal poststenotic dilatation. AB - We present a case of pulmonic stenosis with large aneurysmal poststenotic dilatation that was safely and effectively treated with balloon valvuloplasty. Though the poststenotic dilatation persists after the procedure, the risk of dissection and rupture is very low. Hence, balloon valvuloplasty should be considered the treatment of choice in this setting. PMID- 11870947 TI - Postoperative coronary revascularization on LVAD support for surgically inaccessible myocardial ischemia. AB - The authors have used the concept of hybrid revascularization to salvage a patient in persistent cardiogenic shock after incomplete emergent surgical revascularization. While the patient was on left ventricular assist device support, a complex angioplasty was done on a surgically inaccessible right coronary artery, with subsequent cardiac recovery. PMID- 11870948 TI - Left ventricular aneurysm as a consequence of hypertrophic obstructive cardiomyopathy. PMID- 11870949 TI - The ever fascinating hypertrophic obstructive cardiomyopathy. PMID- 11870950 TI - Incomplete retention after direct myocardial injection. AB - Direct intramyocardial injection may permit local delivery of protein and gene therapy agents for myocardial and coronary artery disease. Little is known about the immediate fate of materials administered via percutaneous endomyocardial catheters or via surgical epicardial injection. In this study, we use a novel method to evaluate the acute retention of agents injected directly into the myocardium, compare epicardial with the percutaneous endocardial and postmortem delivery, and evaluate the influence of injectate volume on myocardial retention. Fifteen 40-50 kg pigs underwent overlapping myocardial injections using a percutaneous endomyocardial catheter, an epicardial needle via an open chest, and epicardial needle postmortem. Multiple distinct 15 micro neutron-activated microsphere species were used as tracers. Two or three myocardial walls were injected in each animal using 3.5 mm, 27-28 gauge needles at varying injectate volumes. Animals were sacrificed immediately. Myocardial walls were divided and multiple microsphere species were quantified. In an additional study, nine 70 kg pigs underwent percutaneous endomyocardial injections with replication-deficient adenovirus encoding for the production of lac-Z. The injectate volume was varied, while the viral particle number remained constant. The animals were sacrificed 5 days after the percutaneous injections; the heart, liver, and spleen were collected for beta-galactosidase activity. Endomyocardial injection was associated with 43% +/- 15% microsphere retention, compared with 15% +/- 21% (P < 0.01) retention of open chest epicardial injection and 89% +/- 60% (P < 0.01) for postmortem injection. Reducing the injectate volume from 100 to 10 microL improved microsphere retention (P = 0.01). There was a trend toward improved viral transfection associated with smaller injection volumes. Despite direct intramyocardial administration, a significant fraction of injectate is not retained locally. Catheter-based needle endomyocardial injection is associated with equivalent or superior injectate retention compared with open chest epicardial injection. Proportionately, more injectate may be retained at lower volumes. Loss may involve a combination of channel leakage, venous, and lymphatic return. PMID- 11870951 TI - CO(2) angiography. AB - Iodinated contrast agents are routinely used in procedures to diagnose and treat peripheral vascular disease. Despite the development of low-osmolar contrast agents and premedication techniques, these agents are still associated with contrast-induced nephropathy and allergic reactions in some individuals. To overcome these problems, carbon dioxide angiography has been developed as an alternative to standard iodinated contrast angiography in certain patient populations. The technology of digital subtraction angiography has greatly improved the image quality of CO(2) angiography. Understanding the unique properties of CO(2), the techniques for its use, and its associated limitations and complications will allow interventional cardiologists to expand their treatments of atherosclerotic peripheral vascular disease. PMID- 11870952 TI - Stent-based antirestenotic coatings (sirolimus/paclitaxel). PMID- 11870955 TI - Effect of bismuth subgallate (local hemostatic agent) on wound healing in rats. Histological and histometric findings. AB - The aim of this research was to evaluate the effect of bismuth subgallate on wound healing. In 40 Wistar rats, two standard wounds (3.5 mm x 2 mm) were made using a biopsy punch on the back of each animal. Test wounds were filled with bismuth subgallate and control wounds with 0.9% saline. At 1, 4, 7, 11 and 18 days, the qualitative evolution of the granulation tissue morphology was observed and digitalized histologic images were evaluated. There were no significant histological differences between test and control. Histometrically, there were statistically significant differences between test and control (ANOVA--days 1 and 4; Student t test, p < 0.05--days 7, 11 and 18) in terms of the following parameters: area of ulceration--day 1; distance between epithelial edges--day 4; area of granulation tissue--days 7, 11 and 18. It was concluded that bismuth subgallate is biocompatible to the healing tissue, and did not interfere with the normal development of wound healing. PMID- 11870957 TI - Reaction of rat connective tissue to implanted dentin tubes filled with a white mineral trioxide aggregate. AB - The purpose of this paper was to study the reaction of rat subcutaneous connective tissue to the implantation of dentin tubes filled with white mineral trioxide aggregate (MTA), a material that will be marketed. The tubes were implanted into rat subcutaneous tissue and the animals were sacrificed after 7 and 30 days. The undecalcified pieces were prepared for histological analysis with polarized light and von Kossa technique for mineralized tissues. Granulations birefringent to polarized light and an irregular structure like a bridge were observed next to the material; both were von Kossa positive. Also, in the dentin wall tubules a layer of birefringent granulations was observed. The results were similar to those reported for gray MTA, indicating that the mechanisms of action of the white and gray MTA are similar. PMID- 11870953 TI - Drug-eluting stents for the prevention of restenosis: in quest for the Holy Grail. PMID- 11870956 TI - Calcium hydroxide root canal dressing. Histopathological evaluation of periapical repair at different time periods. AB - The objective of this study was to evaluate periapical and apical repair using calcium hydroxide root canal dressings for different lengths of times in teeth with induced chronic periapical lesions. A total of 61 root canals of maxillary and mandibular premolars from 4 dogs were used. After mechanical preparation of the root canals using the crown-down technique, and 5.25% NaOCl as irrigating solution, the apical foramen was enlarged in all cases. A calcium hydroxide root canal dressing was applied. The control group did not receive a root canal dressing. The animals were killed at 7, 15 or 30 days. After histological preparation, serial sections were stained with hematoxylin-eosin and Mallory's trichrome. The best histopathological results occurred at 15 and 30 days, and the worst results occurred at 7 days and in the control group. PMID- 11870958 TI - Cariogenicity of different types of milk: an experimental study using animal model. AB - This study evaluated the cariogenic potential of infant formulas and cow's milk, using a high cariogenic challenge in the animal model. Sixty female Wistar rats infected with Streptococcus sobrinus and desalivated were randomly divided into 6 groups, which received ad libitum: 1) sterilized deionized distilled water (SDW) with 5% sucrose; 2) cow's milk; 3) Nan 2; 4) Nestogeno 2; 5) Ninho growth supporting; 6) SDW. Groups 1 and 6 also received essential diet NCP#2 by gavage, twice a day. After 21 days, the animals were killed and evaluated according to recovered oral microbiota and caries score by using a modified Keyes method. The analysis of the carbohydrates in the milk samples was performed using HPLC. The results were analyzed by Shapiro-Wilk and Kruskal-Wallis tests. Cow's milk had the lowest cariogenic potential compared to the other test groups, but it was not statistically different from group 6. The percentage of S. sobrinus obtained from the oral cavity of the animals was not statistically different among the groups studied, except for the SDW group. It was concluded that cow's milk was not cariogenic and infant formulas showed some cariogenic potential. PMID- 11870959 TI - Effects of nicotine on the healing of extraction sockets in rats. A histological study. AB - The aim of the present study was to evaluate, histologically, the influence of nicotine on the socket healing after tooth extraction in rats. Eighty animals were divided into 4 groups of 20 rats each (2 control and 2 test groups). In the first and second test groups, the animals received one dose of nicotine hemisulfate solution once or twice daily, respectively. In the control groups, the animals received sterile saline once or twice daily. All solutions were injected subcutaneously on the dorsum of the animals for 4 weeks. The results showed that nicotine delayed alveolar healing, especially in terms of organization of connective tissue and osteoneogenesis. Angiogenesis was considerably impaired in the ossification area and in the gingival tissues as well. We concluded that the impairment of the healing of extraction sockets was found to be directly related to the drug dosage. PMID- 11870960 TI - Scanning electron microscopy evaluation of the interface of three adhesive systems. AB - The objective of this research was to investigate the resin-dentin interface of three adhesive systems, Scotchbond Multi-Purpose, Optibond and Denthesive Bond II by scanning electron microscopy. The adhesives and their respective composite resins were applied inside the cervical root canal of human incisors and canines according to manufacturer recommendations. The teeth were embedded in acrylic resin and sliced transversally to the root canal and perpendicularly to the resin dentin interface. The adhesive systems Scotchbond Multi-Purpose and Optibond had a homogenous hybrid layer and similar characteristics, involving resin penetration of peritubular and intertubular dentin matrix. Morphological differences of resin tags were seen; Scotchbond Multi-Purpose had more and longer tags than Optibond. Denthesive Bond II did not have the same consistency of bonding. Tubular orifices were not opened and the smear layer was not removed. This was due to the absence of previous acid conditioning of dentin that damages hybrid layer formation. Analysis of the hybrid layer revealed different patterns, suggesting that the attachment was influenced by many factors and a standardization of dentinal substrate was impossible. PMID- 11870962 TI - Effectiveness of an oral hygiene program for Brazilian orphans. AB - The aim of this study was to examine the effectiveness of a preventive oral hygiene program in a group of 7-11-year-old children living in an orphanage in Brazil. The program was based on professional tooth cleaning, as well as dental health information and oral hygiene instruction during a 6-month period. A total of 80 children were examined and 42 who had all first molars erupted were selected for the study. Clinical measurements were recorded at baseline and after 3 and 6 months. Assessment of the efficacy of the program was based on plaque and gingivitis. At the final examination, the mean percentage of surfaces without visible plaque was 36.2% in the experimental group and 15.1% in the control group. These values were also reflected in improved gingival health. The test group showed bleeding upon probing from less than 20% of their interproximal areas, compared to 50% in the control group (p < 0.01). The results of this study indicate improved oral health through the implementation of preventive programs among children who have never been exposed to preventive dental treatment and who are living under adverse social conditions. PMID- 11870961 TI - Fluoride release from restorative materials coated with an adhesive. AB - The retention of both fluoride resins and resin-modified glass ionomer cements to dental tissues can be improved by the association of an adhesive system which promotes the bonding between the resin component and dentin, forming a hybrid layer. The purpose of this investigation was to evaluate if the presence of the adhesive, being part of the hybrid layer composition, interfered with the fluoride released to tooth tissues. The restorative materials studied were: Vitremer (3M), Heliomolar (Vivadent) and Z100 (3M) using an adhesive application (Scotch Bond MultiPurpose Plus--3M). Ten discs of each material were prepared: 5 were covered with the adhesive and 5 were not. The discs were immersed in individual flasks containing artificial saliva which was changed daily. Fluoride release was measured at days 1, 5, 10, 15 and 20 by a fluoride combined electrode (9609 BN--Orion) coupled to an ion analyzer (SA-720 Procyon). One-way ANOVA and the Student-Newman-Keuls test were applied to compare the materials. The results showed that the use of a dental adhesive significantly decreased the fluoride release of Vitremer and reduced the fluoride release of Heliomolar to undetectable levels with the methodology used. PMID- 11870963 TI - Systemic and oral alterations in Brazilian patients with cutaneous herpes zoster. AB - Herpes zoster (HZ) is a virotic disease caused by Herpesvirus varicellae. The objective of this study was to determine the factors that trigger the disease, and the systemic and oral alterations present in Brazilian patients with herpes zoster. A total of 30 patients with HZ and 100 control patients with other diseases were studied. Of the 30 patients with HZ, 13 were male (43.3%) and 17 were female (56.7%), with an average age of 43.2 (range 3-78). The patients were submitted to general clinical, dermatological and intraoral examinations. Only 50% of the HZ patients reported emotional stress at the onset of the disease. A total of 3.7% of the patients were positive for HIV and 11.1% for systemic malignant neoplasm. Cutaneous lesions were found on the thorax (68.3%), face (20%), lower limbs (10%) and upper limbs (6.7%). Specific oral involvement such as oral HZ was not found. The presence of the disease may indicate a non diagnosed malignant neoplasm and/or association with AIDS. PMID- 11870964 TI - Root canal filling with calcium hydroxide using different techniques. AB - Several different techniques for placing calcium hydroxide into root canals have been proposed, such as amalgam carriers, vertical pluggers, McSpadden compactors, Lentulo drills, files and special syringes. Because correct filling of the root canal is necessary for calcium hydroxide paste to act effectively, the aim of this research was to study different techniques of root canal filling with calcium hydroxide pastes in dog teeth. The placement of calcium hydroxide with a file, absorbent paper points and vertical pluggers presented the lowest number of empty spaces in the three thirds of the root canal, followed by the Lentulo drill and the McSpadden compactor. PMID- 11870965 TI - Effect of commercial acrylic resins on dimensional accuracy of the maxillary denture base. AB - Denture base dimensional changes were found with commercial thermocured acrylic resins. Fifteen stone cast-wax base sets were packed for routine flasking. Classico, Vipcril, and Meliodent Multicryl acrylic resins were prepared according to manufacturer instructions. After final acrylic resin pressing, the flasks were placed in strain clamps and submitted to polymerization cycles in heat-water following manufacturer instructions. The resin bases were fixed on casts with instant adhesive and the sets laterally sectioned in the corresponding regions to the distal of canines (A), mesial of first molars (B), and posterior palatal zone (C). The gap between the stone cast and resin base was verified with a measuring microscope at five reference positions for each type of cut. The data were submitted to ANOVA and Tukey's test and a significant statistical difference between the commercial acrylic resins was shown. The pattern of distortion verified in the posterior palatal region was confirmed in the C cut with significant statistical differences among the three acrylic resins. However, there were no significant statistical differences in A and B cuts. PMID- 11870966 TI - Technical assessment of WHO-621 periodontal probe made in Brazil. AB - This study technically assessed the WHO-621 periodontal probe made in Brazil as well as its dimensional and weight stability after sterilization. The following characteristics were measured in one hundred (100) new probes: diameter and linear area of tip sphere and beginning and ending references of its colored band by means of indirect computerized morphometric method, and weight, by means of a digital balance within 0.01 mm precision. Fifty (50) probes underwent one hundred (100) cycles of either steam or heat sterilization. The WHO-621 Trinity probe showed mean values of 0.66 mm in its longitudinal and 0.54 mm in its transversal sphere diameters; 0.40 mm2 in its sphere linear area; 3.56 mm and 5.76 mm for the beginning and ending of the colored band, respectively. Its mean weight was 10.12 g with small variation. Some of the measurements showed significant changes after both sterilization procedures. Neither of the sterilization methods showed to be more advantageous (Student's t-test, p = 0.05). PMID- 11870967 TI - Hyalinizing clear cell carcinoma of minor salivary gland: case report. AB - Hyalinizing clear cell carcinoma is a low-grade neoplasm of the minor salivary gland composed exclusively of epithelial cells and not myoepithelial cells. It predominantly affects the oral cavity of adult females. It is microscopically characterized by hyalinizing stroma and clear cells, which are typically positive for cytokeratin markers and negative for S 100 and smooth muscle actin (SMA). Cystic degeneration can also be present. Pathologists should be aware of this new entity so as not to misdiagnose otherwise. To our knowledge, this is the first case report of its kind from Malaysia. PMID- 11870968 TI - From student to leader. How NSNA makes the difference. PMID- 11870969 TI - Make the most of your NSNA experience. PMID- 11870970 TI - The rewards of starting an NSNA chapter. Student and faculty perspectives. PMID- 11870971 TI - Leadership development and NSNA. PMID- 11870972 TI - The value of mentoring. PMID- 11870973 TI - Finding the right job for you. PMID- 11870974 TI - Interviewing for success. PMID- 11870975 TI - The first six months of practice: strategies for success. PMID- 11870976 TI - Career spotlight. Infusion nursing. PMID- 11870977 TI - Professionalism and activism: one and the same. PMID- 11870978 TI - Forging positive student/faculty relationships. PMID- 11870979 TI - Is the fast track to a higher degree in nursing for you? PMID- 11870980 TI - The critical path to leadership development. A student perspective. PMID- 11870981 TI - Getting the most out of clinicals. PMID- 11870983 TI - The NSNA Leadership U. Learning without walls. PMID- 11870982 TI - A formula for academic success. PMID- 11870984 TI - The center for AIDS and humanity: remembering the past, honoring the heroes, working for a future without AIDS. PMID- 11870985 TI - The ten most oratorically challenged people of 2001. PMID- 11870986 TI - Nutrient deficiency associated with new HIV medication. PMID- 11870987 TI - Gilead sciences releases Viread. PMID- 11870989 TI - 2002: the year ahead. PMID- 11870990 TI - Healthy choices = healthy lives continues to educate the community. PMID- 11870988 TI - A new year of possibilities. PMID- 11870992 TI - ASP joins in MLK March. PMID- 11870993 TI - Come out for the legislative AIDS awareness day. PMID- 11870991 TI - Out with the old, in with the new. PMID- 11870994 TI - Stick-with-it-ness. PMID- 11870995 TI - 2001 NATAF Conference overview. PMID- 11870996 TI - ICAAC study on resistance. PMID- 11870997 TI - Dreams and nightmares. PMID- 11870998 TI - Federal budget passes--finally. PMID- 11870999 TI - Voices united: Asia, Africa and the Americas benefit scheduled for February 28. PMID- 11871000 TI - The black church week of prayer for the healing of AIDS and other upcoming events. PMID- 11871001 TI - Social opportunities in the Metro Altanta HIV/AIDS community. PMID- 11871002 TI - Men, milk cartons and microbicides. PMID- 11871003 TI - Reference accuracy in neonatal-maternal nursing literature. AB - PURPOSE: To identify the frequency and types of reference errors in neonatal maternal nursing literature. This study was an extension of earlier research on reference accuracy in pediatric and critical care nursing journals. DESIGN: A random sample was selected of references in three nursing journals: Neonatal Network: The Journal of Neonatal Nursing; Journal of Obstetric, Gynecologic, and Neonatal Nursing; and The American Journal of Maternal/Child Nursing. References were compared against original publications, and errors were classified as major or minor based on criteria used in earlier studies. RESULTS: Fifty-four of the 221 references had errors, for an overall error rate of 24.4 percent. Major errors were found in 21.3 percent of the references, and minor errors were calculated at 3.2 percent. Errors in the author's name were most common, followed by errors in titles of articles and books. The rates of reference errors in this study were lower than those reported previously in the nursing and medical literature. PMID- 11871004 TI - Accidental administration of oxytocin to a premature infant. AB - Oxytocin has been used for several decades in close proximity to newborns, yet no published information is available regarding complications associated with its accidental administration to a newborn. We describe a case where oxytocin instead of vitamin K was accidentally administered intramuscularly to a premature infant shortly after birth. The patient described remained hemodynamically stable but developed transient hyponatremia as the sole biochemical abnormality. PMID- 11871005 TI - Brazilian mothers' experiences of home care for their low birth weight infants. AB - PURPOSE: To describe Brazilian mothers' experiences of home care for their low birth weight (LBW) infants. DESIGN: Descriptive. SAMPLE: Eleven Brazilian mothers whose LBW infants were hospitalized at birth and followed up for a period of four weeks. MAIN OUTCOME VARIABLE: Brazilian mothers' recollections about the home care of their infants. RESULTS: The study identified the infants' and mothers' profiles and the mothers' concerns about disruption in attachment due to hospitalization and their infants' growth. After hospital discharge, these mothers mainly worried about their infants' weight gain, breathing, and overall development, as well as breastfeeding, feeding, and medication preparation. PMID- 11871006 TI - Noise in the intensive care nursery. PMID- 11871007 TI - Cerebral palsy and the NICU graduate. PMID- 11871008 TI - Posthemorrhagic hydrocephalus: a complication of intraventricular hemorrhage. PMID- 11871009 TI - Osteomyelitis in the neonate. AB - Osteomyelitis is defined as a bacterial infection of the musculoskeletal system. Osteomyelitis in the newborn is relatively rare, but if missed can have devastating, lifelong consequences for the growing infant. Those of us who care for neonates in intensive care units should have an understanding of this disease and be equipped to recognize it and begin treatment quickly to prevent long-term sequelae. This article reviews the pathophysiology of osteomyelitis and explains why the neonate is at an extraordinary risk for this disease. It reviews risk factors, clinical presentation, etiology and pathophysiology, diagnostic evaluation, treatment and monitoring of treatment, and long-term outcomes. A case study of a 27-week gestational age infant who presented with osteomyelitis and septic hip is presented, looking at the infant's hospital course and outcome. PMID- 11871010 TI - [Rational antibiotic therapy and evidence-based medicine]. PMID- 11871011 TI - [A fourth generation cephalosporin--cefepime (maxipime) in pediatrics]. PMID- 11871012 TI - [Cefepime (maxipine) in treating abdominal surgical infections]. PMID- 11871013 TI - [Use of maxipime in treating infections in neutropenic patients]. PMID- 11871014 TI - [Fourth generation cephalosporins in treating severe hospital infections: pharmacoeconomic aspects]. PMID- 11871015 TI - [Modern role of cephalosporin antibiotics for treating infections in the hospital]. PMID- 11871016 TI - [Problems in diagnosing and antibiotic therapy of hospital-acquired pneumonia]. PMID- 11871017 TI - [Experience in using the fourth generation cephalosporin cefepime (maxipime) in treating Russian patients with severe infections]. PMID- 11871018 TI - [Treatment of acute experimental diphtheria toxemia by immunohemosorption]. AB - The antitoxic antidiphtheric immunosorbent DATs was tested in experiments on mice and guinea-pigs using the model of acute diphtheric toxemia. In extracorporeal connecting with the circulation system of the animals the sorbent effectively bound and eliminated from the circulation molecules of diphtheric toxin and antitoxin, reduced significantly their levels in the blood after hemoperfusion. The effectiveness of the immunosorbent DATs was confirmed when it saved the ginea pigs after injection of lethal toxin doses. PMID- 11871019 TI - [Change in some biological properties of cerebrospinal fluid and blood in substance abuse patients during treatment by cerebrospinal fluid-sorption]. PMID- 11871020 TI - [Systemic analysis of a material substrate for development of brain edema swelling]. AB - The biological system behavior was analysed by physical parameters: moisture content and density of brain tissue in biological modeling of compression brain edema-swelling (BES). The systemic analysis objectively assessed material substrate of the process pathogenetic mechanisms. Main shifts of such mechanisms exists in the intact brain and start to operate in response to the provoking factors. As a result, the system "brain tissue" rapidly and progressively loses its internal reserves. One can permanently trace the ambi-trends in the system by the factors interhemispheric and intrahemispheric "physical parameters". PMID- 11871021 TI - [Metabolic shifts in stress caused by hunger, and pharmacotherapy of them by a new taurine derivative]. AB - The influence of a new N-phenylalkyl derivative of taurine in combination with a diet enriched with soybean proteins was studied in rats with nutritional dystrophia induced by 3/4 reduced standard diet. It is shown biochemically and morphologically that usage of the drug simultaneously with the diet stimulated protein synthesis, reduced lipid peroxidation, normalized spermatogenesis and compensatory adaptation of different functions. PMID- 11871022 TI - [Blood system response to emotional stress and low-dose ionizing radiation]. AB - Rat experiments have shown that long-term emotional stress provokes adaptive reaction consisting in temporary activation of all the bone marrow lineages. Gamma preradiation (2 days before the stress) in low doses (0.5 and 0.2 Gy) destroyed this adaptive reaction dose-dependently: 0.5 Gy preradiation arrested activation of all the lineages while 0.2 Gy preradiation prevented activation of only a lymphoid lineage. It is inferred that low-dose ionizing preradiation influences hemopoietic system of stressed animals. PMID- 11871023 TI - [Interconnection of immunologic indicators of blood and functional parameters of leukocytes in children with relapsing herpesvirus infection]. AB - Correlations between immunological indices of blood and functional activity of leukocytes were studied in healthy children and those with recurrent Herpes virus infection (RHVI) at the age 3-6 and 7-11 years. Functional activity of leukocytes was assessed by the chemiluminescence test. Both healthy and RHVI children are shown to have age-specific features of blood immunology, leukocyte function and their correlation. In ontogenesis and RHVI relations between immunological indices of blood and functional parameters of leukocytes vary and correlations appear between leukocyte function and cellular, humoral immunity. These findings evidence for differentiation of ontogenetic processes in different links of immunological system, on the one side, and necessitate development of normal age standards which will facilitate diagnosis of immunoreactive disorders in various infections, on the other side. PMID- 11871024 TI - [Effect of blocking angiotensin receptors on cerebral blood flow in the post ischemic period in hypertensive rats]. AB - Acute experiments on anesthetized rats have been conducted to study the effect of lozartan (5 mg/kg i.p.) on cerebral circulation and its autoregulation in hypertensive animals (a vasorenal model) in postischemic period. Transitory ischemia was induced by bilateral compression of the carotid arteries for 12 minutes, continued--by ligation of the carotid arteries. It was found that blockade of angiotensin receptors by lazartan provoked lowering of systemic arterial pressure. Changes in cerebral circulation were unstable, depended on arterial pressure and vascular cerebral resistance. The lower limit of autoregulation of the brain flow shifted to higher arterial pressure in hypertensive animals before brain ischemia and disorder after ischemia. Lozartan promoted recovery of the autoregulatory reactions of the brain vessels and raised survival of rats in continuous ligation of the carotid arteries. PMID- 11871025 TI - [Effect of regulatory peptides on the effect of cardiorespiratory synchronization in patients with heart diseases]. AB - A long-acting somatostatin analogue (sandostatin) and synthetic opioid dalargin were examined for the effect on the heart rhythm under the cardiorespiratory synchronization test in patients with heart disease. In high-frequency respiration synchronized with photostimulator flashes the patients exhibited cardiorespiratory synchronism when heart rate corresponds exactly to respiration rate. A spontaneous change of the respiration rate entailed corresponding changes in the heart rate. This allowed regulation of the heart rhythm. Intravenous injection of sandostatin or dalargin extended the range of the cardiorespiratory synchronization. The latter points to involvement of peptides in mechanisms of heart taking of the control signals coming from the central nervous system. PMID- 11871026 TI - [Experimental model of peptic stomach ulcer]. PMID- 11871027 TI - [Stress phenomenon. Emotional stress and its role in pathology (end)]. PMID- 11871028 TI - [Neuroimmunoendocrine interactions: their role in dysregulatory pathologies]. AB - The author reviews the trends in development of views of interaction of the basic regulating systems (nervous, endocrine and immune) supporting control and adaptive functions of the brain. Two historical periods are singled out: the period of neuroendocrinology and neuroimmunoendocrinology. Mechanisms of interaction of brain regulating systems and various autoimmune diseases resulting from defects in the above mechanisms are considered. Development of cooperative interactions of the above regulating systems at early evolutionary stages is suggested. PMID- 11871029 TI - [Disruption of acquired head and paw movement coordination after unilateral lesioning of the motor cortex in dogs (a kinematic analysis)]. AB - Dogs were trained to remove a cup with meat to the head bent down to the feeder and hold the limb flexed during eating. At the early stage of learning, the stable innate head-forelimb coordination characteristic for untrained animals was manifest. The forelimb flexion was accompanied by anticipatory lifting of the bent head, and the following bending of the head led to an extension of the flexed forelimb. The opposite coordination, i.e., the lifting and holding of the forelimb when the head is bent down, was achieved only by training. The lesion of the motor cortex contralateral to the working forelimb in the trained dogs led to a prolonged disturbance of the simultaneous holding of the flexed forelimb and the head bent down. The lesion of the motor cortex did not affect the individual movements but disturbed their coordination. In the operated dogs the innate relationships between the head and forelimb movement recovered. The results support the previous finding that the lesion of the motor cortex led to recovery of the innate coordination transformed in the process of learning. PMID- 11871030 TI - [Selective attention in dogs from energy characteristics of neocortical potentials in the 1-220 Hz band]. AB - The state of selective attention was studied in dogs in the course of instrumental conditioning. During interstimuli intervals, this state was manifested in the state of strained waiting for conditioned stimuli. Electrical activity of different areas in both hemispheres was analyzed using the Fast Fourier Transformation. It was shown that in the process of development of selective attention, the high-frequency EEG components (40-200 Hz) in the motor area of the right hemisphere and the visual and parietal areas of the left hemisphere had a predominant significance over the traditional EEG frequencies of 1-30 Hz. The state of selective attention was characterized by another functional mosaic organization of the neocortical potentials. PMID- 11871031 TI - [Various resistance of motor preference in rats exposed to forced retraining]. AB - Adult Wistar rats were trained to get food from a narrow tube under conditions of free choice of a limb. After reaching a stable level of using the preferred limb, rats (n = 35) were forced to relearn the acquired skill for reaching food with a unpreferred paw. When afterwards the rats returned to the initial conditions of free choice of a limb, 12 animals (34%) did not recover their initial preference, 8 rats (23%) were ambidextrous, and 15 animals (43%) returned to the originally preferred paw. The results demonstrated different resistance of the initial limb preference to forced retraining in different animals. It is suggested that the process of retraining per se is not the only and sufficient condition of the change in "handedness". It also suggested that the different resistance of initial preference to forced retraining reflects individual differences in a degree (intensity) of this preference, which are predetermined by internal, probably by genetic factors. PMID- 11871032 TI - [Administration of aggregated beta-amyloid peptide (25-35) causes change in long term potentiation in the hippocampus]. AB - A month after intracerebroventricular injection of aggregated beta-amyloid fragment 25-35 (7.5 nmol/ventricle), a significant change in in vivo long-term potentiation in hippocampus was demonstrated. The time course of the long-term potentiation was compared with that in sham-operated animals, a powerful and stable increase in the evoked potential amplitude was observed. This phenomenon can be related with the oxidative stress that was revealed in this model in our previous studies, and, as a consequence, with deterioration of ion homeostasis. PMID- 11871033 TI - [Dynamics of local shifts in oscillations and energy metabolism of the rabbit cerebral cortex during formation of a conditioned defensive reflex]. AB - Brain energy metabolism in different functional states or activities of humans and animals is characterized by dynamic changes in the degree of coupling between glycolysis and tissue respiration in different cell compartments (Fox et al., 1988; Fox, 1989; Pellerin et Magistretti, 1994; Prichard et al., 1991; Schur et al., 1999). These processes determine variations in the brain redox state (Siesjo, 1978) that can be potentiometrically recorded with implanted platinum electrodes as the brain tissue redox state potential E (Puppi et Fely, 1983). This potential was recorded in rat brain cortex with four pairs of platinum electrodes implanted into different symmetrical cortical region (one electrode of a pair being located in the cortical layers, another being located epidurally). In the course of defensive conditioning (after 5-15 combination of a bulb light and a weak electrodermal stimulation of a ear), E oscillations (6-10 per minute) appeared. In this period, stimuli combinations produced the generalized E shifts. Later on (with accumulation of stimuli combinations), the episodes of E increase were replaced by the episodes of E decrease. To the 200-400th combinations, E oscillations disappeared, and E shifts became local and stable. The findings suggest that conditioning shifts the balance between the main energy-producing systems in the brain tissue: at the initial stages of conditioning brain functions are predominantly supported by the energy obtained from tissue respiration, while during the realization of defensive conditioning glycolysis is the main source of energy. PMID- 11871034 TI - [Participation of 5-HT1A-receptors in regulating various types of aggressive behavior]. AB - We studied the effects of selective agonists of 5-HT1A receptors 8-OH-DPAT and flesinoxan on aggressive behavior of C57BL/6 male mice in the "resident-intruder" test and on defensive aggression of Norway rats toward man. 8-OH-DPAT (0.4 mg/kg, i.p.) significantly reduced the intermale aggression in mice and defensive aggression in rats (0.1-0.5 mg/kg, i.p.). In the dose of 0.5 mg/kg, flesinoxan inhibited the aggressive behavior in mice. These results suggest that activation of 5-HT1A receptors reduces different kinds of affective aggression. The results are discussed in terms of interaction between the well-known anxiolytic effects of 5-HT1A agonists and their antiaggressive properties. PMID- 11871035 TI - [Possible mechanisms for forming typological features of behavior of WAG/Rij line rats]. AB - Typological behavior reactions of WAG/Rij rats were studied from the standpoint of divergent modulatory integration hypothesis. This rat strain has a genetically determined dominant dysfunction of the benzodiazepine system of the thalamic nuclei. This disorder provokes an epileptiform disease such as absence epilepsy. It was suggested that the dysfunction of this system would result in a modification of the modulatory systems, which support the motivation states of escape and avoidance reactions as well as of the modulatory systems, which form the emotional states. Modifications of these states are the background of typological behavioral features of WAG/Rij rats. It was shown that WAG/Rij have the lower threshold of the development of haloperidol catalepsy, higher levels of fear and depression. On the first day of training in a shuttle box, WAG/Rij rats demonstrated better avoidance performance than Wistar rats. On the second and 28th days, the amnestic effect of the epileptiform disease was observed. The amnestic effect was also observed after passive avoidance conditioning. The results are discussed in terms of the modulatory integratin hypothesis. PMID- 11871036 TI - [Phase-dependent coordination of two motor programs in the buccal ganglion of a pteropod mollusk]. AB - Rhythmic activities of two feeding structures of the pteropod mollusk Clione limacina, redula and hooks, controlled by the neural networks in the buccal ganglia must be coordinated in order to produce a meaningful feeding response. Optical recording from the buccal ganglia, which allows the simultaneous activities of numerous neurons to be traced, revealed that such coordination exists in a phase-dependent manner. Instead of recording four theoretically possible phases of neuronal rhythmic activity, we always recorded only two phases, even after the electrical stimulation of the cerebro-buccal connective, which triggers both radula and hook rhythmic movements in the preparation. PMID- 11871037 TI - [Participation of Na/Ca-exchange and intracellular mobilized Ca2+ in regulating depression of cholino-sensitive Helix lucorum neurons to a cellular analog of habituation]. AB - The role the Na/Ca-exchange and intracellular Ca2+ released from Ca(2+)-depots in the modulatory action of Na,K-pump inhibitor ouabain on cholinosensitivity in the command neurons of Helix lucorum was studied in a cellular analogue of habituation. The integral transmembrane inward currents in LPa2, LPa3, RPa3, and RPa2 neurons were recorded in Helix lucorum ganglia preparation using two electrode voltage clamp technique. The reduction of cholinosensitivity of a neuron was estimated as a depth of the depression of the acetylcholine-induced inward currents during the rhythmic local acetylcholine applications (with the interstimulus interval of 2-4 min) on a somatic membrane. The inhibitor of the Na/Ca-exchange benzamil (the extracellular action, 15-35 mcM) and two specific inhibitors of Ca-ATPase in the sarcoplasmic and endoplasmic reticulum, cyclopiazonic acid and thapsigargin (intracellular injection by spontaneous diffusion, 0.1 mM) prevented the modification of the depression of acetylcholine induced current by ouabain (100 mcM) during the rhythmic application of acetylcholine. A conclusion is drawn that the inhibitor of the Na,K-pump ouabain modifies the depression of neuron cholinosensitivity in the cellular analogue of habituation via the Na/Ca-exchange and intracellular Ca2+ released from Ca2+ depots. PMID- 11871039 TI - [Effect of mexidol on status of conditioned reflex activity after traumatic brain damage]. AB - Even partial hippocampal lesions in rats resulted in a disturbance of time interval determination over the course of several months (1200-1500 presentations) other complex conditioned reactions being preserved. As distinct from the control animals, the long period of failure of time interval counting was absent in rats receiving Mexidol. Continuous time conditioning took place in these animals. Due to substantial improvement of autonomic processes and emotional reactions, it was possible to present a higher number of conditioned stimuli in experiments. Mexidol seems to improve the compensatory and recovery processes after brain injuries: the impaired functions recover faster, the rate of the retrograde degeneration in the lesioned brain structures decreases, phenomena like Monakov's diaschis are not observed etc. PMID- 11871038 TI - [Behavioral analysis of consequences of chronic blockade of NMDA-type glutamate receptors in the early postnatal period in rats]. AB - In view of the hypothesis that glutamatergic dysfunction of brain can underlie the negative symptoms of schizophrenia (including cognitive deficit), the aim of this study was to develop a model of cognitive impairment in Wistar male rats after administration of a noncompetitive NMDA-receptor antagonist in early postnatal period. Rat pups were daily subcutaneously injected with 0.05 mg/kg MK 801 on postnatal days 7-49. On the 27th and 28th days 24 h after the last previous injection, the MK-801-treated rats demonstrated lower spontaneous locomotor and exploratory activity in comparison with saline control, however, they retained the reaction of hyperlocomotion which developed immediately after the MK-801 administration. In these rats, the anxiety level in the elevated plus maze (on the 40th postnatal day) was found to be decreased, and the spatial learning in food rewarded task was negatively affected (on the 50th-54th days). It is suggested that impairment of the input of sensory information and its correct assessment by the animals can be associated with the early neonatal blockade of NMDA glutamate receptors. PMID- 11871040 TI - [Variability of heart rhythm in dynamic study of the psychovegetative relationship in neurogenic syncope]. AB - Physiological changes accompanying syncopes of neural origin (SNO) in patients with psychovegetative syndrome are still insufficiently studied. The data concerning the role of the autonomic nervous system are discrepant. Heart rate variability was analyzed in 68 patients with SNO in a supine position and during the active 20-min orthostatic test taking into account the heart rate components of very low frequency (VLF, an index of cerebral sympathetic activity) and high frequency (HF, a marker of vagal modulation). Steady growth of the VLF and progressive decrease in the LF within 15-20 min of the orthostasis were observed in all the patients (n = 33), who fainted after this period. The predominance of the VLF in the heart rate power spectra was correlated with a high level of anxiety. It is suggested that this fact indicates the stable cerebral sympathetic activation resulting in a baroreceptor dysfunction, i.e., a failure of vasomotor regulation in patients with SNO. PMID- 11871041 TI - [Activity of neurons in the anterolateral region of the rabbit motor cortex during taking food and nonfood objects in instrumental behavior]. AB - We showed earlier that only 2-4% of N-neurons in the rabbit's anterolateral ("masticatory") motor cortex (AC) that are being specialized in relation to Newly formed acts (such as pressing the pedal, approaching the pedal) during the elaboration of instrumental food-acquistion behavior. The majority of neurons in this area are O-neurons that are specialized in relation to the Older acts formed long before the acquisition of instrumental behavior: mostly taking of food. It was shown also that electrical stimulation of this area produced jaw movements. It might seem reasonable to propose that the number of N-neurons may be increased in this area if pedal pressing in substituted with instrumental act like seizure and pulling a ring that is more corresponding to the "projectional property" of the AC. Here we show that the number of N-neurons in rabbit's AC remains constant -3.4%, even is the animals that were trained to pull a ring instead of pressing a pedal. Thus, we assume that the small number of N-neurons is the stable property of the AC. However, the role of the AC in the subserving of the behavior does not remain the same. Much more AC units (O-neurons) were activated in the ring pulling than in the pedal pressing instrumental act. PMID- 11871042 TI - [Distribution of behaviorally specialized neurons and expression of the c-fos transcription factor in the rat cerebral cortex during learning]. AB - In order to investigate the relation between the immediate early gene induction in neurons and neuronal specialization in respect to functional systems of newly formed behavior, the neuronal expression of Fos transcription factor in rat brain was studied in animals of learning group and home cage control group. Animals of the learning group acquired a new behavior of pressing a pedal. Fos-positive neurons were counted in the retrosplenial area of cingulate cortex and anterolateral area of motor cortex, i.e., the two brain regions that differ in the number of neurons showing specific activity during this behavior. In the home cage control animals the number of Fos-positive neurons was low and no difference was found between the two brain regions. In the animals of the learning group the number of Fos-positive neurons was significantly higher in the cingulate cortex characterized by the greater number of neurons specialized in relation to the system of new behavior. These findings suggest that the c-Fox expression can indicate the processes of neuronal specialization. PMID- 11871043 TI - [Modified water maze for studying spatial memory of rats]. AB - A method for evaluation of impairments of spatial memory in rats is put forward. Sensitivity of the advanced method based on the principles of Morris' water maze was compared with that of the classic prototype. Efficiency of the advanced method was assessed by dose- and time-dependent effects of agroclavine on the spatial memory of rats. Agroclavine (10 micrograms/kg) was shown to produce spatial memory impairment in rats. The obtained results also indicate that the modernized maze is more sensitive in revealing impairments of the spatial memory in rats than the classic water maze. PMID- 11871044 TI - Organ recovery from a donor with end-stage renal disease: a case study. AB - As the field of transplantation enters the new millennium, maximizing organs per donor remains one of the greatest challenges of procurement. This case study outlines nontraditional medical management techniques that facilitated the recovery of 5 transplantable organs from a patient with end-stage renal disease. Strategies utilized in this case included the use of continuous veno-venous hemodialysis and airway pressure release ventilation to maximize the outcomes of the donation. Although the use of these strategies is admittedly limited to hospitals where the resources are available, this case study suggests that utilizing available resources in any clinical setting can make more organs available to people on the waiting list for solid organ transplantation. PMID- 11871045 TI - Living donors' perception of their quality of health after donation. AB - To alleviate the long wait on the cadaveric transplant list, recipients are pursuing the option of living donation. Potential donors may have significant concerns about the decision to donate, including the quality of health after donation. This study identifies living donors' perception regarding their quality of health after donating a kidney, and identifies the perception of the adequacy of the medical follow-up after donation. In this retrospective study, the 36-Item Short Form Health Survey was mailed to all kidney donors at the transplant center from January 1995 to December 1998 (n = 118). The study population scored significantly better than the US sample population in all 8 categories of the questionnaire. When comparing scores using the relationship of the donor to the recipient, mean scores of donors who were "distant relatives" were found to be lower in 7 of the 8 categories. Most donors who had follow-up after donation felt it to be excellent to good; yet 50% stated they had no healthcare follow-up after donation. Themes were identified regarding the type of follow-up donors felt they needed after donation. Donors perceived their quality of health after donation as better than the general US population. Donor follow-up needs to be emphasized and followed more systematically. PMID- 11871046 TI - St John's wort: a hidden risk for transplant patients. AB - Herbal medications may cause prescription drug interactions in transplant recipients. After 2 of our kidney transplant recipients started self-medicating with St John's wort, their cyclosporine concentrations were consistently documented to be subtherapeutic. While on St John's wort, one patient developed acute rejection possibly due to low cyclosporine concentrations. Termination of St John's wort returned both patients' cyclosporine concentrations to therapeutic values. Based on the Naranjo Adverse Drug Reaction Probability Scale, our report would achieve a "probable" score, which supports the existence of a St John's wort-cyclosporine adverse drug interaction. St John's wort may induce cytochrome P-450 3A4 activity and/or P-glycoprotein expression, which are both involved in the metabolism and absorption of cyclosporine. Patients using St John's wort concomitantly with cyclosporine or other medications with similar absorption and/or metabolism to cyclosporine need close monitoring. Transplant coordinators are in a critical position to educate transplant recipients about the potential risks of herbal medication usage. PMID- 11871047 TI - Psychosocial and behavioral selection criteria for solid organ transplantation. AB - An evidence-based selection process for organ transplantation may be a valuable approach to improve posttransplant outcomes. This paper reviews state-of-the-art psychosocial and behavioral selection criteria and assesses their validity in view of predicting outcomes after transplantation. Psychosocial factors addressed are psychiatric disorders, mental retardation, irreversible cognitive dysfunction, and lack of social support. Behavioral selection criteria discussed are alcoholism, smoking, drug abuse, and obesity. This review reveals that the evidence concerning these selection criteria in scarce. There is a definite need for more longitudinal research to strengthen the scientific basis of the psychosocial and behavioral dimension of transplantation. PMID- 11871048 TI - Use of structured self-monitoring in transplant education. AB - A self-assessment instrument for use at home by transplant recipients was developed to help foster partnership between patients and their healthcare provider. Self-monitoring at home has not replaced the need for close follow-up but does allow patients to provide concrete data to their healthcare provider in order to promote earlier detection of and response to adverse events. Patients are taught the essentials of self-monitoring while they are in the hospital for their transplant. Patients who perform routine self-assessment would be able to detect and provide information about problems early in the course of events. Thus, early intervention could potentially decrease the severity of the problem and prevent repeated hospitalizations. The concern that patients would not be able to perform a reliable self-assessment was unfounded; patients exceeded expectations and embraced the opportunity to communicate physical signs and symptoms effectively. PMID- 11871049 TI - Use of behavioral contingency contracting with heart transplant candidates. AB - For many years, behavioral contingency contracting has been useful in increasing compliance in a variety of patient populations. More recently, this type of contracting has been used to facilitate communication and motivate behavioral change among patients referred for transplantation. This article discusses the theoretical basis of behavioral contingency contracting and describes the development and implementation of a multipurpose behavioral contract for patients receiving heart transplants. The scarcity of donor organs and the stringent posttransplant regimen mandates that psychosocial as well as medical variables be considered during the transplant evaluation process. Behavioral contingency contracting is useful in identifying those individuals with the potential to maintain a transplanted organ capably. PMID- 11871050 TI - Overcoming therapeutic misconceptions with patient education. PMID- 11871051 TI - Psychosocial sequelae of a heart-transplant recipient with a histrionic personality disorder. PMID- 11871052 TI - Donor Action: an international initiative to alleviate organ shortage. AB - CONTEXT: Donor Action, an international initiative to alleviate organ shortage, provides a comprehensive state-of-the-art methodology that helps critical care units develop a tailor-made approach to optimize donation practices and performance. OBJECTIVE: To report the impact of the Donor Action methodology on organ donation rates in 8 countries (70 critical care units) in North America and Europe. DESIGN: Baseline data on the clinical potential for donation, staff attitudes, knowledge toward donation, self-reported confidence in performing a range of donation roles, and educational requirements were gathered. These data were analyzed using the Donor Action database and improvement measures were introduced to address identified weaknesses. RESULTS: Following introduction of the program's improvement measures, which addressed identified weaknesses, donations increased on average by 53% (P = .0017) per country at 1 year. Sustained improvements settled at 70% to 160% increases at 3 years. Although Donor Action is at various stages of implementation in different countries, the number of centers and countries demonstrating an immediate awareness effect is increasing and sustained effects in centers with the longest follow-up promise a significant impact on donation rates as more countries implement this methodology. PMID- 11871053 TI - Attitudes about organ and tissue donation among the general public and blood donors in Hong Kong. AB - CONTEXT: The cadaveric organ and tissue donation rate in Hong Kong is not satisfactory; 1 million blood donors are registered and more than 300,000 are active. However, the current attitudes toward organ and tissue donation in the general public and blood donors of Hong Kong are unknown. METHODS: Random general public (n = 1018) and blood donors (n = 1227) of Chinese origin, with age ranging from 16 to 60 years, were interviewed using a standard verified questionnaire that examines attitudes and knowledge of organ and tissue donation. RESULTS: The mean age of the general public and blood donors were 32.6 and 28.9 years, respectively. Of the general public, 44.4% were men and among blood donors, 60% were men. About 56% of both groups thought that organ donation is an obligation of citizens. Blood donors were more aware than the general public about the types of organs that can be donated. When compared with the general public, a significantly higher percentage of blood donors were willing to donate their organs (81% vs 53%), had heard about organ donation cards (98.3% vs 89.5%), and had signed the cards (49.9% vs 22.6%). About 70% of both groups who had signed a card were carrying it. Thirty-nine percent of the general public and 17% of blood donors had not decided whether they would donate. For blood donors, 49.7% were willing to donate their relatives' organs, compared with 41.8% of the general public. Most individuals in both groups would not object to their relatives' decision to donate. About two thirds of individuals in both groups disagreed with the concept of an opt-out law, though only 20% of the general public and 14.4% of blood donors would refuse donation if an opt-out law were in practice. CONCLUSIONS: This study shows that blood donors have better knowledge of organ donation and are more willing to donate their organs and sign an organ donation card than the general public. However, a substantial proportion of blood donors have not signed a donor card. It would be useful to design promotion programs to facilitate blood donors' participation in organ donation. PMID- 11871055 TI - [Structure of trophic insufficiency in wounded persons with traumatic disease]. PMID- 11871054 TI - [Prospects for prognosis in cardiac surgery: experience from using discriminant linear analysis]. AB - The authors evaluate the possibility of using the discriminant linear analysis in predicting the unfavourable outcomes at aortocoronary bypass. There were investigated 98 patients (the mean age 64.4 +/- 3.0 years) who underwent the surgical revascularization of myocardium in 1992-1997. 56 signs characterizing the demographic, clinical-and-instrumental and operational indices were analyzed. The obtained discriminant equation of operation outcome prediction included 15 signs. The operation for unstable angina, the cholesterol level and the patient's age were the most informative parameters. The total per cent of correct classification was 87.8%. According to the authors' experience the discriminant linear analysis is the informative method of multi-dimensional statistical analysis permitting to reveal the most significant signs that influence on the prognosis of surgical intervention. PMID- 11871056 TI - [Serotonin level in endocrine cells in bronchial mucosa and stomach in chronic obstructive lung diseases]. AB - 85 patients aged 26-60 years with pulmonary chronic obstructive diseases (PCOD) and gastric and duodenal erosive-ulcerous lesions were investigated. In PCOD the increased bioamines level in endocrine cells of bronchial ciliated epithelium and gastric glandular epithelium was detected before the treatment. The complex treatment in the form of bronchoscopy with lymphotropic administration of antibiotic and immunomodulator against the background of intragastral ozonotherapy contributes to the decrease in tissue edema, restoration of their trophicity and inflammatory process resolution. Against the background of therapy the serotonin content in endocrine cells of bronchial ciliated epithelium and gastric mucosa decreased to the norm. PMID- 11871057 TI - [Current approaches to managing patients with extra-hospital pneumonia in military collectives]. PMID- 11871058 TI - [Clinical and methodical aspects of monitoring blood pressure]. PMID- 11871059 TI - [Hemosinus and severe cranial-cerebral trauma]. PMID- 11871060 TI - [Treatment and evacuation characteristics of sanitary losses from the effect of neurotoxicants]. PMID- 11871062 TI - [Features of cardiovascular pathology in pilots participating in remediation after the accident at Chernobyl Atomic Energy Station]. PMID- 11871063 TI - [A new formulary for drugs]. PMID- 11871061 TI - [Interferon inducers in treating acute respiratory diseases: problems and prospects]. PMID- 11871064 TI - [On unknown medals with portrayals of medical service colonel-general A. A. Vishnevsky]. PMID- 11871065 TI - [Formation and development of domestic neurology at the Military-Medical Academy]. PMID- 11871066 TI - [Second Central polyclinic of the RF MD is 80 years old ]. PMID- 11871067 TI - [The Department of Military and Extreme Medicine of Russian State Medical University is 80 years old]. PMID- 11871068 TI - [Prospects for using dermatoglyphics for evaluating the mental status of service members]. AB - The authors discuss the perspectives of practical use of morphological method for investigation of psychical diseases and predisposition to them. The method is based on the analysis of picture of finger and palm papillary pattern dermatoglyphs. This marker doesn't change during the life and the authors attribute it to one of the most important directions in professional selection of specialists of military professions. PMID- 11871069 TI - [Astigmatism after trans-scleral posterior chamber lens implantation in young children]. AB - BACKGROUND: The purpose of this study was to evaluate the postoperative astigmatism after trans-scleral fixation of intraocular lenses in children and to develop a strategy for amblyopia prophylaxis. PATIENTS AND METHODS: In eight eyes with lentectomy, posterior chamber intraocular lenses were fixed in the ciliary sulcus by trans-scleral sutures. The postoperative astigmatism was measured by retinoscopy every 1 to 2 weeks in the first 2 months, followed by monthly intervals thereafter. Keratometry was performed with an automated hand keratometer. RESULTS: Postoperative astigmatism was 2.5 to 8.0 D. The astigmatism regressed to 0.75 D in half the eyes within 4 weeks and in five of the eyes within 16 weeks. In one of the eight eyes, the astigmatism decreased to 2.0 D and in two it remained unchanged. The astigmatism did not change any more after 16 weeks postoperatively. Visual acuity was 0.016 to 1.0. CONCLUSIONS: Our study shows that the postoperative astigmatism regresses soon and does not seriously interfere with amblyopia therapy. If greater than 2 D, half of the astigmatism should be corrected with glasses, even in the early postoperative period. After 16 weeks, full correction is recommended. PMID- 11871070 TI - [Eye contusions caused by a bottle cap. A retrospective study based on the Erlangen Ocular Contusion Register (EOCR)]. AB - BACKGROUND: Perforating ocular injuries due to exploding beverage bottles are well known. However, bottle caps alone may also induce severe ocular damage due to ocular contusions while opening carbon dioxide-containing beverage bottles. We studied this type of ocular contusion. PATIENTS AND METHODS: Retrospectively, we examined all findings, operations, and tension profiles of 400 consecutive patients with ocular contusion who had been hospitalized in the Department of Ophthalmology, University Erlangen-Nurnberg, using the Erlangen ocular contusion registry (EOCR). RESULTS: Seven patients were injured by bottle caps (1.8%) while attempting to open a beverage bottle. The bottle caps were screw caps in six patients and crown corks in one patient. The patient age was 34 +/- 22 years (range: 16-68 years). Four patients were female, three male. We found the following morphological changes: hyphema (seven patients), iridodialysis (two patients), traumatic cataract (two patients), vitreous prolapse (one patient), vitreous hemorrhage (two patients), and retinal detachment (two patients). Intraocular pressure was higher than 21 mmHg in three patients. Visual acuity was hand motion in two patients, lower then 0.4 in two, and 1.0 in three on the admission day. Five patients read 1.0 on the day of discharge. CONCLUSIONS: Bottle cap injuries are not as harmless as supposed. Bottle caps may induce severe ocular damage due to the high-impact energy. PMID- 11871071 TI - [Ocular toxoplasmosis antibodies in aqueous humor and serum]. AB - BACKGROUND: The diagnosis of ocular toxoplasmosis is mainly based on ophthalmological examination but might be difficult to establish in some cases. The purpose of our study was to evaluate the value of aqueous humor and serum analysis in ocular toxoplasmosis. PATIENTS AND METHODS: We analyzed the avidity of toxoplasma-specific IgG in aqueous humor and serum samples from 50 patients with toxoplasmic retinochoroiditis, with 25 patients with uveitis posterior or panuveitis serving as controls. RESULTS: Specific intraocular antibody synthesis could be confirmed in 49 patients (98%). In two patients (8%) of the control group, antibody synthesis was detected (false positive). Forty-nine patients with diagnoses of ocular toxoplasmosis were positive for serum anti-T. gondii IgG, but only three patients had increased IgM levels. CONCLUSIONS: Analysis of local antibody production is a reliable method for confirming or excluding a suspected clinical diagnosis of toxoplasma retinochoroiditis. The determination of toxoplasma antibodies in the patients' serum is of limited value. PMID- 11871073 TI - [Ophthalmological manifestations in neurofibromatosis. Diffuse choroid involvement in von Recklinghausen neurofibromatosis type I]. PMID- 11871072 TI - [Dacryocystostomy and microsurgical lacrimal sac reconstruction in dacryolithiasis]. AB - BACKGROUND: The incidence of dacryoliths ranges between 8 and 14% of patients undergoing a dacryocystorhinostomy. Lacrimal duct stenoses with dacryoliths can be managed either by surgery or minimally invasive techniques. We present two patients who underwent removal of dacryoliths by dacryocystotomy and microsurgical reconstruction without osteotomy. PATIENTS AND METHODS: Case report 1: A 47-year-old male patient presented with chronic epiphora. The dacryocystogram revealed a filling defect in the dilated lacrimal sac. Case report 2: A 22-year-old female patient suffered from a hemorrhagic right-sided dacryocystitis. Two years previously the patient had undergone lacrimal endoscopy with silicone intubation for 3 months. Preoperative dacryocystography showed an obliteration of the superior portion of the lacrimal sac. In both patients, we performed the extraction of the dacryolith by dacryocystotomy followed by silicone tube intubation of the lacrimal system for 3 months. Both patients with a follow-up of 24 and 29 months, respectively, showed good clinical results and patent nasolacrimal pathways on digital dacryocystography. RESULTS: Methods to extract dacryoliths include minimally invasive techniques such as nonsurgical radiologic or endoscopic removal during dacryoplasty or surgical approaches such as external or endonasal dacryocystorhinostomy. CONCLUSION: Dacryocystotomy with primary microsurgical lacrimal sac reconstruction combined with silicone intubation is an alternative surgical procedure to remove dacryoliths. An osteotomy is not always required. PMID- 11871074 TI - [Papillary plica/caruncle tumor. Squamous epithelial carcinoma of the conjunctiva associated with human papillomavirus type 16]. PMID- 11871075 TI - [The myopia puzzle. Disorders in fine tuning of length and focal distance of the eye]. PMID- 11871076 TI - [Innovative surgery of glaucoma. New goals are established]. PMID- 11871077 TI - [Perspectives in glaucoma surgery]. AB - Trabeculectomy is still considered to be the gold standard in the surgical treatment of the open-angle glaucomas. The additional application of local antimetabolites has reduced the rate of early filtering bled fibrosis, but increased the rate of essential late-postoperative complications. Growth factor inhibition and photodynamic therapy may be an alternative local treatment to enhance the results in filtering surgery. Non-penetrating glaucoma surgery and ab interno trabecular surgery have several conceptual advantages, e.g. the lack of overfiltration or the untouched conjunctiva in the ab-interno approach. Clinical studies of these and other new procedures including antiglaucomatous retinectomy and subchoroidal shunt systems are currently performed in order to evaluate their potential and limits in the clinical management of glaucoma. PMID- 11871078 TI - [Pigment epithelium detachments in AMD (age-associated macular degeneration) and "polypoid choroidal vasculopathy". A fluorescein and indocyanine green angiography study]. AB - AIMS: A prospective analysis to differentiate the angiographic appearance of pigment epithelial detachments (PED) in age-related macular degeneration (AMD) was performed. The aim was to verify if characteristics of polypoidal choroidal vasculopathy (PCV) can be observed in elderly patients with Central European background. PATIENTS AND METHODS: Fluorescein and indocyanine green angiography was performed in 101 consecutive patients (53-87 years, 63 females, 38 males) with clinical signs of PED and drusen. RESULTS: Different types of PED could be differentiated: PCV-associated PED in 14 patients (13.9%), vascularized PED in 72 patients (71.2%), and nonvascularized PED in 15 patients (14.9%). CONCLUSIONS: The clinical diagnosis of PED in AMD can be differentiated by angiography into PCV-associated PED, vascularized PED, and nonvascularized PED. This differentiation is important because PCV-associated PED may have a pathogenetic and genetic background other than age-related changes in Bruch's membrane. These patients also may share a natural course and treatment options different from PED in AMD. PMID- 11871079 TI - [Osteo-odonto-keratoprosthesis. A summary of 6 years surgical experience]. AB - PURPOSE: In patients with severe corneal scarring and vascularisation, corneal grafts have a high risk of failure. In those patients an osteo-odonto keratoprosthesis (OOKP) can be performed. METHOD: We performed OOKP in eight patients in all with ocular pemphigoid, severe alkali burns, or repulsion of a corneal graft. Each of the patients had finger-counting visual acuity or less. The OOKP consisted of a PMMA cylinder 8 mm long, with a diameter of 3-4 mm, and from a root of the patient's tooth. This was implanted in the cornea and covered with buccal mucosa. RESULTS: The medium follow-up was 3 years. The implants are now well incorporated. The best visual acuity was between 0.6 and 0.9 in four patients with intact retinas. In the remaining patients, visual acuity was dependent on the posterior segment findings. The visual field was centred. Five of the eight patients had vitreous bleeding after the operation that was spontaneously absorbed. Revision of the mucous coverage was necessary in four patients, and one of them had a severe inflammation with partial absorption of the bone and tooth implant. One patient developed a secondary angle-closure glaucoma and was treated successfully by implantation of an Ahmed valve. Two patients developed membranes behind the cylinder that were successfully removed. There was no loss of any prosthesis during follow-up. CONCLUSIONS: In impasse in severe corneal scars, the osteo-odonto-keratoprosthesis is still indicated and shows good results in visual rehabilitation of patients. PMID- 11871080 TI - [Changes in neovascular membranes and normal choroid blood vessels after multiple photodynamic therapy treatments]. AB - PURPOSE: ICG angiography (ICGA) was used to document the effect of repeated PDT (verteporfin) on size and leakage of choroidal neovascularisation in age-related macular degeneration (AMD) and treatment-related side effects on the choroid. METHODS: Forty-two patients were followed over 24 months in a clinical trial for PDT in AMD. The ICGAs were performed every 3 months with a confocal laser scanning system. Patients received repeated verteporfin treatment. At each control visit, the patients were retreated if leakage was present in fluorescein angiography (FA). RESULTS: A continuous, highly significant reduction in CNV size and leakage area was found over 24 months. The initial CNV size dropped by 23% from 3.86 mm2 to 2.98 mm2. The leakage area in the late phase of the angiogram decreased by 30.3% from 5.0 mm2 to 3.5 mm2. A significant side effect of PDT on the choroid was documented by an increased hypofluorescent area in ICGA. The maximum size of the hypofluorescent area was reached after 12 months. At month 24, the choroidal fluorescence showed recovery in respect to area and intensity of fluorescence. But hypofluorescence surrounding the CNV lesion was already present in 40 out of 42 eyes before treatment. CONCLUSION: The ICGA confirms that repeated PDT treatments lead to a significant reduction in CNV size and leakage area over as long as 2 years. CNV lesions are surrounded by choriocapillary hypofluorescence in ICGA. PDT causes further hypoperfusion of the choroid but in the long-term significant recovery of choroidal perfusion was shown. PMID- 11871081 TI - [Recent findings in studies of post-traumatic stress disorder]. PMID- 11871082 TI - [Various ventilation modes and management of patients under mechanical ventilation]. PMID- 11871083 TI - [Cardiopulmonary bypass and blood coagulation]. PMID- 11871084 TI - [Recent progress in therapy of diabetes mellitus]. PMID- 11871085 TI - [Epidural opioids]. PMID- 11871086 TI - [International guidelines for cardiopulmonary resuscitation]. PMID- 11871087 TI - [Depth of anesthesia monitored with Bispectral Index]. PMID- 11871088 TI - [Diagnosis and therapy of patients with malignant hyperthermia]. PMID- 11871089 TI - [New muscle relaxants]. PMID- 11871090 TI - [Perioperative anesthetic management of abdominal organs]. PMID- 11871091 TI - [Fundamental and clinical studies on osmotic pressure]. PMID- 11871092 TI - [Perioperative anesthetic management in neurosurgery]. PMID- 11871093 TI - [Molecular mechanism of biological rhythm]. PMID- 11871094 TI - [Anesthesia for organ transplantations--special reference to pharmacokinetics of anesthetics and other agents used during perioperative period]. PMID- 11871096 TI - [Apoptosis signals and anesthesia]. PMID- 11871095 TI - [Alpha2 agonists and their clinical use in anesthesia]. PMID- 11871097 TI - [Allergic reactions during anesthesia]. PMID- 11871098 TI - [Necessity of routine preoperative laboratory and diagnostic screening]. PMID- 11871099 TI - [Infections and NO]. PMID- 11871100 TI - [Transplantation and immunity]. PMID- 11871101 TI - [Crisis management for high-pressure gases during disasters--experience from the Hanshin earthquake]. PMID- 11871102 TI - [Pathophysiology of bronchial asthma--special reference to the role of airway epithelial cells]. PMID- 11871103 TI - [Physiopathology of sleep apnea syndrome]. PMID- 11871104 TI - [Regulatory mechanism of neurotransmitter release by low-molecular-weight G protein, Rab3A and the regulatory proteins]. PMID- 11871106 TI - [Recent findings in studies of hypertension therapy]. PMID- 11871105 TI - [Anesthetics and their drug interactions]. PMID- 11871107 TI - [Recent findings on local anesthetics]. PMID- 11871108 TI - [Fundamental and clinical studies on intravenous anesthesia]. PMID- 11871109 TI - [Inhalation anesthetics and heart functions]. PMID- 11871110 TI - [Updates on perioperative management of body temperature]. PMID- 11871112 TI - [Laughter, the brain, and psychiatric diseases]. PMID- 11871111 TI - [Laughter and the mind--the mystery of laughter]. PMID- 11871113 TI - [The fourth revision of the Medical Treatment Law and future problems]. PMID- 11871114 TI - [Functional classification of psychiatric wards]. PMID- 11871115 TI - [Functional classification of psychiatric wards--on practice of treatment of drug dependence]. PMID- 11871116 TI - [Behavioral restriction and its minimization in psychiatric practice]. PMID- 11871117 TI - [Drug therapy of organic psychosis]. PMID- 11871118 TI - [Sleep disorders among the aged and their treatment]. PMID- 11871119 TI - [Development of the mind: from developmental psychology to neuroscience]. PMID- 11871120 TI - [The Japanese Pure Land (Jodo) concept of Buddhism and Naikan (introspection) therapy]. PMID- 11871121 TI - [Change of myosin heavy chain isoform expression and histocytochemical findings in skeletal muscular atrophy due to simulated weightlessness by hindlimb tenotomy]. PMID- 11871122 TI - [Surgery in rats]. PMID- 11871123 TI - [Surgery in cats and experimental methods]. PMID- 11871125 TI - Overview of studies on rat sperm motion analysis using a Hamilton-Thorne Sperm Analyzer--collaborative working study. AB - This collaborative study was conducted to determine the utility and sensitivity of nine sperm motion parameters generated by a Hamilton-Thorne Sperm Analyzer (HTM-IVOS) for detecting adverse effects of chemicals on sperm motion in rats. The efficacy of sperm motion parameters was investigated using nine reproductive toxicants: adriamycin, alpha-chlorohydrin (3 different studies were carried out), dinoseb, ethylene glycol monoethyl ether, 2,5-hexanedione, sulfasalazine, trimethyl phosphate, and ornidazole. The percentage of motile sperm (% motile sperm), the only parameter expressing the status of semen containing non-motile sperm, detected adverse effects on sperm motion in 9 out of 10 studies. However, weak effects on sperm motion were not detected by this parameter in 4 out of 7 studies in which sperm motion disorders were noted at medium or low dosages. The percentage of progressively motile sperm (% progressive sperm) and the sperm velocity parameters (average path velocity, straight line velocity, and curvilinear velocity) detected adverse effects on sperm motion in all studies. In 7 studies which noted sperm motion disorders at medium or low dosages, weak effects on sperm motion were detected by the % progressive sperm in 5 studies and by the sperm velocity parameters in 6 studies. In 10 studies, amplitude of lateral head displacement (ALH) did not detect adverse effects on sperm motion in 4 studies, and beat cross frequency (BCF) failed to detect adverse effects on sperm motion in 3 studies. Because ALH and BCF show the swimming pattern of spermatozoa as head movement, the characteristics of these parameters are different from the % progressive sperm and the sperm velocity parameters. Straightness (STR) and linearity (LIN), which are secondary parameters calculated from sperm velocity parameters, could not detect adverse effects on sperm motion when the sperm velocity parameters did not detect adverse effects. On the basis of these results, we concluded that the % progressive sperm and sperm velocity parameters are useful and sensitive indicators for detecting adverse effects on sperm motion. However, in the % progressive sperm, setting up a suitable threshold of VAP and/or STR is important to gain further sensitivity for detecting adverse effects on sperm motion. The % motile sperm is useful for assessment of sperm motion disorder, and ALH and BCF are useful for evaluating the swimming pattern of sperm. STR and LIN are not very useful for detecting adverse effects on sperm motion. PMID- 11871124 TI - Peroxisome proliferator-activated receptor alpha (PPAR alpha) agonist, WY-14,643, increased transcription of myosin light chain-2 in cardiomyocytes. AB - Peroxisome proliferator-activated receptors (PPARs) are nuclear hormone receptors that can be activated by xenobiotics and natural fatty acids. To assess the potential physiological activity of PPAR ligands on cardiac muscular cells, the effects of PPAR alpha agonist, WY-14,643, on both rat hearts and a rat cardiomyocyte cell line (H9c2 cells) were investigated. Male F344 rats were fed a diet containing WY-14,643 at a concentration of 100 ppm for 26 weeks. Cardiac muscular hypertrophy was revealed by morphometric analysis in which the diameter of the muscular fibers in WY-14,643-treated rats was larger than those of control rats. Using H9c2 cells in vitro, the protein content per cell was increased in a dose-dependent manner with the treatment of WY-14,643. The transcription of myosin light chain-2 (MLC-2), a parameter of myocardial hypertrophy, was increased in H9c2 cells transfected with the rat MLC-2/luciferase fusion gene by WY-14,643 as well as other peroxisome proliferators, clofibrate and di(2 ethylhexyl) phthalate. In addition, accumulation of myosin light chain protein was confirmed in H9c2 cells treated with WY-14,643 at 10 micrograms/ml for 7 days or more by immunohistochemistry. These results suggest that PPAR alpha ligands have a potential to regulate MLC-2, which is a contractile protein in cardiomyocytes and may play a part of role in the pathogenesis of cardiac hypertrophy. PMID- 11871127 TI - Investigation of usefulness of sperm analyses in dogs for male fertility study. AB - The usefulness of sperm analyses in dogs, including sperm motion analysis, was investigated. Sperm motion analysis was performed with the CellSoft-4000 computer assisted sperm analysis (CASA) system. First, we examined the conditions for preservation of optimal semen quality. We found that sperm retained more of their motility at 4 degrees C than at 37 degrees C. Secondly, we observed sperm motion, concentration and morphology in dog semen continuously for 11 weeks. We collected semen samples during the test period, and the samples retained sperm motion, concentration and morphology. Finally, we administrated alpha-chlorohydrin, which decreases rodent sperm motion, at a single oral dose of 100 mg/kg or 150 mg/kg to dogs. Sperm motion was inhibited immediately after alpha-chlorohydrin treatment, and recovered after 2 weeks. None of the experimental animals were sacrificed in the above-mentioned examinations. We thus confirmed that sperm analyses including motion analysis in dogs are useful in male fertility studies. PMID- 11871126 TI - Comparative study of toxicity of 4-nitrophenol and 2,4-dinitrophenol in newborn and young rats. AB - The toxicities of 4-nitrophenol and 2,4-dinitrophenol in newborn and young rats was examined and the susceptibility of newborn rats was analyzed in terms of presumed unequivocally toxic and no observed adverse effect levels (NOAELs). In the 18-day repeated dose newborn rat study, 4-nitrophenol was orally given from Day 4 to Day 21 after birth but did not induce any toxicity up to 160 mg/kg in the main study, although it induced death in one of six males at 160 mg/kg, and three of six males and one of six females at 230 mg/kg in a prior dose-finding study. In the 28-day repeated dose oral toxicity study starting at 6 weeks of age, 4-nitrophenol caused the death of most males and females at 1,000 mg/kg but was not toxic at 400 mg/kg except for male rat-specific renal toxicity. As unequivocally toxic levels were considered to be 230 mg/kg/day in newborn rats and 600 to 800 mg/kg/day in young rats, and NOAELs were 110 mg/kg/day in newborn rats and 400 mg/kg/day in young rats, the susceptibility of the newborn to 4 nitrophenol appears to be 2.5 to 4 times higher than that of young animals. In the newborn rat study of 2,4-dinitrophenol, animals died at 30 mg/kg in the dose finding study and significant lowering of body and organ weights was observed at 20 mg/kg in the main study. In the 28-day young rat study, clear toxic signs followed by death occurred at 80 mg/kg but there was no definitive toxicity at 20 mg/kg. As unequivocally toxic levels and NOAELs were considered to be 30 and 10 mg/kg/day in newborn rats and 80 and 20 mg/kg/day in young rats, respectively, the toxicity of 2,4-dinitrophenol in newborns again seems to be 2 to 3 times stronger than in young rats. Abnormalities of external development and reflex ontogeny in the newborn were not observed with either chemical. Based on these results, it can be concluded that the toxic response in newborn rats is at most 4 times higher than that in young rats, at least in the cases of 4-nitrophenol and 2,4-dinitrophenol. PMID- 11871128 TI - Effect of temperature on the frequency of chromosome aberrations and micronuclei in cultured Chinese hamster cells. AB - We previously reported that both hyperthermia and hypothermia induced micronuclei in mouse bone marrow cells (Asanami and Shimono, 1997a, 1997b, 1999). To investigate the effects of temperature on chromosome aberration in vitro, we conducted chromosome aberration and micronucleus tests under hyper- and hypothermic conditions using Chinese hamster lung (CHL) cells. In the chromosome aberration test, we observed positive responses at 40 degrees C and 41 degrees C for 24 hr, and at 42 degrees C for 6 hr and over. In the micronucleus test, we observed positive responses at 31 degrees C, 33 degrees C, and 40 degrees C for 24 hr, and at 42 degrees C for 2 hr. The results suggest that in CHL cells, hypothermic conditions can induce micronuclei while hyperthermic conditions can induce both chromosome aberrations and micronuclei. PMID- 11871129 TI - Dioxins in bile in relation to those in the human liver and blood. AB - The levels of 20 dioxin congeners, 7 polychlorinated dibenzo-p-dioxins (PCDDs), 10 polychlorinated dibenzofurans (PCDFs) and three coplanar polychlorinated biphenyls (Co-PCBs), in bile were examined in comparison with those in the blood and liver, in 27 autopsy cases. Total-TEQ values were the same in the bile (43.2 +/- 30.9 pg TEQ/g lipid) and blood (43.1 +/- 24.2 pg TEQ/g lipid), and three times higher in the liver (127.8 +/- 57.4 pg TEQ/g lipid). Highly chlorinated PCDDs and PCDFs have a tendency to accumulate in the liver, and their levels in bile and blood were relatively low compared with those in the liver, with 1,2,3,4,6,7,8-heptachlorodibenzofuran having the highest tendency among the 20 congeners. Daily excretion in bile was calculated to be 54 pg TEQ at age 65, by assuming that daily bile secretion is 750 ml and is concentrated 7.5-fold in the gallbladder. The correlation between bile and blood total-TEQ was high, with a correlation coefficient of 0.89 among the 27 autopsy cases. Thus, the regression equation of y = 1.14x - 6.02 will provide us levels in bile (total TEQ per g lipid), knowing the blood total-TEQ level, where x is total-TEQ per g lipid of the blood. Furthermore, accumulation of dioxins was estimated to be 0.99, 0.70 and 1.91 pg TEQ/g lipid/year in bile, blood and liver, respectively. PMID- 11871130 TI - Relationship between toxicity and cadmium accumulation in rats given low amounts of cadmium chloride or cadmium-polluted rice for 22 months. AB - To clarify toxic effects of long-term oral administration of low dose cadmium (Cd) on the liver and kidney, six groups of female Sprague-Dawley rats were fed a diet containing Cd-polluted rice or CdCl2 at concentrations up to 40 ppm, and killed after 12, 18, and 22 months. With toxicological parameters, including histopathology, there was no evidence of Cd-related hepato-renal toxicity, despite a slight decrease of mean corpuscular volume and mean corpuscular hemoglobin of red blood cells with 40 ppm CdCl2. Dose-dependent accumulation of Cd was observed in the liver and kidneys with peak levels of 130 +/- 42 micrograms/g and 120 +/- 20 micrograms/g, respectively, at 18 months in animals treated with 40 ppm CdCl2. A dose-dependent increase in urinary Cd levels became evident with time. Induction of metallothionein (MT) was also observed in the liver and kidney with a high correlation to the corresponding Cd levels. In the proximal renal tubular epithelia of 40 ppm CdCl2-treated rats at 22 months, prominent accumulation of Cd was observed in secondary lysosomes associated with MT deposits in their exocytotic residual bodies. The results demonstrated that, in contrast to the case with high-dose Cd-administration, renal toxicity is not induced by long-term oral administration of low amounts of Cd, although tissue accumulation does occur. Possible protective mechanisms may be operating. PMID- 11871131 TI - [Production of recombinant human CRP and its application on clinical testing]. AB - Recently, changes in the serum CRP level 1/10 the concentration range ordinarily used as a marker of acute inflammation has received attention in relation to cardiovascular injury at the AACC/CDC joint forum at Atlanta held on March 13, 2001. We have succeeded in the development of recombinant human CRP(rCRP) by inserting the cloned CRP gene into expression vector pTRP, followed by transformation of E. coli. Genes encoding the signal peptide of E. coli alkaline phosphatase and kil gene were additionally inserted, so that rCRP can be secreted into the culture supernatant. Five grams of rCRP was purified from 180 L culture supernatant by affinity chromatography. The purified rCRP was indistinguishable from native rCRP with respect to Ca(2+)-dependent binding activity to phosphorylcholine, electrophoretic behavior in the presence or absence of SDS, N terminal amino acid, and immunochemical properties. rCRP was found to have a potential as a reference material and/or calibrator for hsCRP assay. PMID- 11871132 TI - [Factors interfering with CRP assay and various methods of producing reagents that avoid some interference factors]. AB - Recently, our customers have demanded highly precise methods of CRP assay, since the significance of quantitative analysis using very low concentration of CRP has been reevaluated. We must very carefully develop additional devices and produce new reagents, in order to avoid some interference factors to CRP assay. We will explain our process for development of reagents and various ideas to avoid such interference factors, as follows. i.e. 1. selection of latex, 2. selection of antibody, 3. selection of the sensitization methods. We will be able to produce high performance reagents that can avoid some interference factors using procedures. However, after launching the reagents, we sometimes receive complaints that a particular reagent may be influenced by unknown factors that were not anticipated the development stage. We will explain the way that we confirmed unknown factors and our efforts to discover the causes of the interference with our products in that case. PMID- 11871133 TI - [Significance of high sensitive CRP assay for early detection of newborn babies infection diseases]. AB - We have evaluated the accuracy of high sensitive CRP assay method using evanescent wave Immunoassy system(Evanet 20) and significance of this assay on the early detection of infectious diseases in newborn babies. In this assay system, prozone phenomenon was not detected up to 40 mg/dl. The reproducibility of this assay was quite good and the intra run CV value of the same sample was less than 5% for the assay of serum, plasma and whole blood. There was a high correlation between the CRP values in the serum and plasma(r = 0.98, regression formula y = 0.89x + 4.07). Similarly, the values in whole blood and serum samples were quite well correlated(r = 0.98, regression formula y = 0.91x - 6.75). Various humoral elements such as bilirubin, hemoglobin and Chyl did not significantly influence this assay method. A slight increase in blood CRP was clearly demonstrated in the early phase of infectious diseases of newborn babies and monitoring of CRP by this assay system seemed to be quite useful to detect the early phase of infectious diseases in newborn babies. This assay system requires only a small quantity of whole blood to perform quantitative analysis of very small amounts of other substances. Accordingly, this assay system seems to be quite effective for monitoring minute increases in various proteinaceous blood components in emergent laboratory examination or POCT. PMID- 11871134 TI - [High-sensitivity C-reactive protein (CRP) assay--a novel method for assessment of risk ratios for atherosclerotic vascular diseases]. AB - CRP has long been used as a sensitive marker for infectious diseases. Since its serum concentration elevates more than 10 mg/dl with gram-negative bacterial infections, the sensitivity can be enough to be around 0.3-0.6 mg/dl for the diagnosis. However, the sensitivity should be higher in the early diagnosis of infections in new-borne babies. In addition, recently, it was suggested that atherosclerotic lesions are a kind of vasculitis, and the information could be transmitted via production of inflammatory cytokines and acute phase proteins. In fact, serum CRP and serum amyloid protein A(SAA) levels are elevated even in patients with coronary atherosclerosis without acute coronary syndrome(ACS). However, the level was much lower than the cut-off for diagnosis of bacterial infection. Therefore, the high-sensitive assay method has been applied. As the result, high-sensitivity(hs) CRP assay was found to be one of the most sensitive markers for prediction of future ACS in USA. Combination of hsCRP and atherogenic index such as total cholesterol/HDL cholesterol or LDL cholesterol/HDL cholesterol ratio is more useful. Similarly, it was found that hsCRP could predict the future prevalence of ACS even in Japan. It may be true because production of CRP is independent upon the genetic backgrounds. Early prevention of ACS by the measurement of hsCRP is calculated to be economic even if we measured hsCRF often in subjects without symptoms, because medical cost for treatment of acute myocardial infarction is enormous. In patients with high risk for coronary heart diseases, hsCRP-guided therapy is possible by using aspirin, stains, and antibiotics for prevention of ACS. PMID- 11871135 TI - [What is bioinformatics?]. AB - Yet bioinformatics has neither clear definition nor proper Japanese equivalent for this word. Bioinformatics is mainly composed of two fields or disciplines; one is computer and information technology as a tool for analyzing massive data such as genome and proteome, and the other one is research for the integrated biology based on the knowledge of genome. Classical and narrow-sense bioinformatics corresponds to the former. The latter called new bioinformatics aims to understand living thing as a whole, where both genome information and mathematical model play complimentary roles. From now the postgenome area on, this new bioinformatics may give novel perspective about living thing, biology and human. PMID- 11871136 TI - [An introduction to the computer-aided structure-based drug design--applications of bioinformatics to drug discovery]. AB - Computer-aided drug design methods are effective tools for drug discoveries. It is only small number of compounds that are required to be synthesized in practice, if the computer-aided drug design methods are used to predict the activities of the compounds. The methods are based on the interaction theory between drugs and their target proteins. Steric, electrostatic and hydrophobic complementarities are important in the interaction between the drug and the target protein. When three-dimensional structures of target proteins are known, the method is called as a structure-based drug design, while in case that the structures are unknown, a ligand-based drug design approach is employed for the computer-aided rational drug design. In the computer-aided structure-based drug design, it is important to build up the three-dimensional structure of drug protein complex and to calculate a binding affinity of the drug to the protein. A molecular dynamics/free energy calculation is one of the most powerful tools to predict the binding affinity of a ligand molecule, when the three-dimensional structure of the drug-protein complex is available. The molecular dynamics/free energy calculation has been able to evaluate the binding affinities of a lot of ligand molecules to the target proteins successfully. PMID- 11871137 TI - [Gene expression profiling using improved SAGE]. AB - Serial analysis of gene expression(SAGE) is an experimental technique designed to gain a direct and quantitative measure of gene expression. The SAGE method is based on the isolation of unique sequence tags(9-10 bp in length) from individual mRNAs and serial concatenation of tags into long DNA molecules for lump-sum sequencing. The SAGE-method can be applied to the studies exploring virtually any kinds of biological phenomena in which changes in cellular transcription are responsible. SAGE is a highly competent technology that can not only give a global gene expression profile of a particular type of cell or tissue, but also facilitate identification a set of specific genes related to the cellular conditions by comparing the profiles constructed for a pair of cells maintained under different conditions. However, the original SAGE method contains several serious intrinsic problems, such as procedural difficulty in library construction, extremely short cDNA tags, and so forth. We therefore modified the procedure to overcome these drawbacks, and applied the improved method to analyze various biological phenomena. In this review, we present an outline of the method, and describe several studies performed using the method as a major strategic tool. PMID- 11871138 TI - [Common diagnostic tests under the Clinical Laboratory Improvement Amendments of 1988 (CLIA '88) in the United States]. AB - The lower quality of laboratory testing in unlicensed physicians' office laboratories(POLs) had led to legislation of the Clinical Laboratory Improvement Amendments of 1988(CLIA '88) in the United States. This legislation extended laboratory regulations for quality control and assurance, personnel qualification, record-keeping, and proficiency testing to all laboratories regardless of size, complexity, or location, including POLs and ancillary testing sites in a hospital. According to the implementation of the CLIA '88 in 1992, all testing sites in this country must have inspections and a certificate issued by the federal government. The CLIA '88 has improved the quality of testing in POLs, forcing office physicians to deal with the problem of laboratory quality management, thereby increasing laboratory costs. Thus, compliance with the CLIA '88 standards is expensive. On-site testing in POLs has been reduced, discontinued, or changed as a result of the CLIA '88 legislation. A number of POLs have closed, and physicians have restricted test menus to those with simpler methodology(waived tests) because waiver laboratories do not require inspections by the government. Large portion of laboratory tests, which were formerly done in POLs, flow into the reference laboratory market as outreach tests. Currently, 77% of POLs are performing only waived tests or tests in the provider-performed microscopy procedures category, while only 23% have a certificate for moderate or high complexity methodology status. Thus, common diagnostic tests performed in POLs are predominantly based on the waived tests, which are largely different from those performed in Japan, with respect to test item and methodology. PMID- 11871139 TI - [An ambulatory respirometer (Hotmate) as a tool for screening of sleep apnea syndrome]. AB - The gold standard diagnostic method for sleep apnea syndrome(SAS) is overnight polysomnography(PSG), but is costly in terms of time and money. We studied the usefulness of a 24-hour ambulatory respirometer equipped with oximeter(Hotmate) for screening of SAS. Seventy-six cases of suspected SAS were enrolled(68 males and 8 females, mean age 51). The correlation between data from Hotmate and PSG was evaluated in 24 cases who underwent both of the tests for the final diagnosis of SAS. There was a good correlation between the two parameters of the data obtained by Hotmate(H) (H-apnea index(AI) vs H-desaturation index(DI)). Among 24 cases who underwent both Hotmate and PSG, there was a good correlation between the data from PSG and Hotmate(PSG-AI vs H-AI: r = 0.80, p < 0.001). Both sensitivity and specificity were highest when screening criteria of H-DI > 15 was utilized(sensitivity = 91.7%, specificity = 66.7%). Our findings suggest that the respiromonitor with oximeter is useful for the screening the patients with SAS. PMID- 11871141 TI - [Arbekacin resistant gene, aacA/aphD, in Staphylococcus aureus is lost during in vitro passage]. AB - We analyzed for the presence of the aacA/aphD gene in arbekacin resistant Staphylococcus aureus. Tested strains were 50 clinically isolated Staphylococcus aureus that had an MIC of more than 2 micrograms/ml for arbekacin. Among the primary cultures, aacA/aphD genes were detected in 42 of 50 strains(84%) overall, including 34 of 41 Methicillin resistant Staphylococcus aureus(MRSA) strains, and 8 of 9 strains of Methicillin sensitive Staphylococcus aureus(MSSA). After 10 or 20 passages, the aacA/aphD gene was lost in 5 strains of MRSA and 2 strains of MSSA. In the strains that lost the gene, the MIC of arbekacin was reduced remarkably in 3 strains of MRSA and 2 strains of MSSA. Further, MICs of arbekacin decreased along with passages even in the strains that did not loose the aacA/aphD gene, as well as in the strains that did not possess the aacA/aphD gene originally. The fact that MIC values are easily reduced with in vitro passage is important to consider when performing the arbekacin-resistance test for Staphylococcus aureus. PMID- 11871140 TI - [Application of HPLC method for the measurement of hyaluronan in pleural fluid--a new alternative of cellulose acetate membrane electrophoresis]. AB - We examined whether high-performance liquid chromatography(HPLC) method was more suitable for the measurement of hyaluronan(HA) in pleural fluid than the standardized electrophoretical method. Forty-four samples of pleural fluid were measured both by the HPLC method and by the electrophoretical method. The HA values measured by the HPLC method did not show significant correlation(r = 0.33) to those measured by the electrophoretical method. We investigated the reason of this insufficiency of the correlation. The electrophoretical method was not specific for the measurement of HA and other glycosaminoglycan, such as chondroitin could be measured by this method. Therefore, the HA value measured by this method was higher than that measured by the HPLC method. The other problem was some loss of HA during the pre-treatment of samples with percloric acid in the electrophoretical method, which was needless in the HPLC method. We concluded that the HPLC method was more appropriate than the electrophoretical method for the measurement of hyaluronan in pleural fluid. PMID- 11871142 TI - [Studies on the prognostic factors of idiopathic acute sensorineural deafness]. AB - Acute sensorineural deafness(ASD) presents with various hearing types, and dizziness and tinnitus are accompanied in some cases. The purpose of this study was to estimate the influence of risk factors, including dizziness and tinnitus, on the recovery of hearing acuity. Eighty-three patients who visited our hospital within 1 month from its onset and who were diagnosed with ASD between October 1998 and September 1999 were retrospectively reviewed. The patients whose etiology was defined, or whose symptoms shifted to inconsistent deafness were excluded. There were 35 males(ages ranged from 22 to 71 years; average 49 years) and 48 females(ages ranged from 17 to 79 years; average 46 years). The improvement rates were analysed using the following factors such as levels and hearing types at initial examination, interval between its onset and treatment, and presence or absence of dizziness and tinnitus. Concerning hearing types at initial examination, tan type or low sound disturbance type resulted in favorable prognosis. The patients whose treatment was started by the 7th day from its onset had significantly better prognosis than those whose treatment begun after 15 days. The patients who were complicated with tinnitus, especially accompanied with dizziness, indicated a significantly worse prognosis than those without tinnitus. Tan type or low sound disturbance type is a favorable factor, and tinnitus is an unfavorable factor for ASD prognosis. Though tinnitus generally is a significant factor for poor prognosis, this study remains to suggest that it alone causes the poor prognosis. The sooner treatment is provided indicates the better outcome. PMID- 11871143 TI - [Detection of serum anti-p53 antibodies in patients with various types of cancer]. AB - The p53 tumor suppressor gene is altered in a variety of human cancers and mutated p53 protein was found to induce anti-p53 antibodies in sera of patients with various types of malignant neoplastic disease. We report here the presence of anti-p53 antibodies in sera from patients with various disorders as well as from healthy controls by enzyme-linked immunosorbent assay. The prevalence of anti-p53 antibodies in cancer was higher than neoplastic disease. High antibody concentrations were found in patients with 1 pharyngeal, 3 esophageal, 4 lung, 4 gastric, 1 pancreatic, 3 colonic and 4 ovarian cancer. Serological analysis does not require biopsy of the tumor. Therefore, determination of anti-p53 antibodies can be used for tumor screening and for early diagnosis. PMID- 11871144 TI - [Comparative test to evaluate routine HDL-C determination--dextran sulfate Mg2+/cholesterol dehydrogenase method]. PMID- 11871145 TI - Power spectral analysis of heart rate variability for assessment of diurnal variation of autonomic nervous activity in guinea pigs. AB - We established characteristics of power spectral analysis of heart rate variability, and assessed the diurnal variations of autonomic nervous function in guinea pigs. For this purpose, an electrocardiogram (ECG) was recorded for 24 hr from conscious and unrestrained guinea pigs using a telemetry system. There were two major spectral components, at low frequency (LF) and high frequency (HF) bands, in the power spectrum of HR variability. On the basis of these data, we defined two frequency bands of interest: LF (0.07-0.7 Hz) and HF (0.7-3.0 Hz). The power of LF was higher than that of HF in the normal guinea pigs. Atropine significantly reduced power at HF. Propranolol also significantly reduced power at LF. Furthermore, the decrease in the parasympathetic mechanism produced by atropine was reflected in a slight increase in the LF/HF ratio. The LF/HF ratio appeared to follow the reductions of sympathetic activity produced by propranolol. Autonomic blockade studies indicated that the HF component reflected parasympathetic activity and the LF/HF ratio seemed to be a convenient index of autonomic balance. Nocturnal patterns, in which the values of heart rate in the dark phase (20:00-06:00) were higher than those in the light phase (06:00-20:00), were observed. However, the HF, LF and the LF/HF ratio showed no daily pattern. These results suggest that the autonomic nervous function in guinea pigs has no clear circadian rhythmicity. Therefore, this information may be useful for future studies concerning the autonomic nervous function in this species. PMID- 11871146 TI - Genetic characterisation of Mus musculus and related forms from India. AB - The present study reports the biochemical genetic profile of commensal forms of Indian wild house mouse populations Mus musculus from ten localities as well as other species of genus Mus, M. platythrix, M. booduga and M. terricolor III. Extreme polymorphism seen by the presence of a large number of variants and novel alleles in these animals indicates that the Indian M. musculus holds considerable potential for development of useful models for biomedical research. PMID- 11871147 TI - Comparative mapping of 31 rat genes. PMID- 11871148 TI - Immunohistochemical demonstration of c-fos protein in neurons of the medulla oblongata of the musk shrew (Suncus murinus) after veratrine administration. AB - We subcutaneously injected 0.5 mg/kg veratrine into the musk shrew (Suncus murinus), observed the presence or absence, latency, and the incidence of vomiting in each animal for 90 min, and selected animals that frequently vomited (FV group) and those that did not vomit (NV group). Subsequently, animal brains were removed, and the induction of c-fos protein (Fos) was immunohistochemically examined to evaluate neuronal activity in the medulla oblongata. The distribution of Fos-positive neurons in the medulla oblongata was similar between FV and NV groups, with numerous neurons along the entire length of the nucleus of the solitary tract and in the ventrolateral reticular formation. Both veratrine injected groups showed higher numbers of positive neurons than the saline administered group. However, while the FV group showed a high concentration of positive neurons in the dorsal-dorsomedial reticular formation of the nucleus ambiguus in the rostral medulla, the NV group showed few positive neurons in this area. Fos activity in neurons in this area appeared to be higher in animals with a higher incidence of vomiting. PMID- 11871149 TI - Meriones meridianus and Lagurus lagurus as alternative definitive hosts of Echinococcus multilocularis and E. granulosus. AB - The utilization of Meriones meridianus and Lagurus lagurus as alternative definitive hosts for Echinococcus multilocularis and E. granulosus was investingated. Tapeworm stage development of E. multilocularis was observed and their recovery rate was determined in the small intestine of M. meridianus and L. lagurus. These were compared with those in golden hamsters, which are known as alternative definitive hosts. The animals were treated with PTBA (prednisolone tertiary butylacetate) and PA (prednisolone acetate), after which M. meridianus showed the highest recovery rate, whereas L. lagurus had few or no worms. The recovery rate of worms from golden hamsters was between those of M. meridianus and L. lagurus. On day 20 post-infection, developing tapeworms with mature segments were collected from M. meridianus treated with PA. The worms were mostly from the proximal and medial small intestine. The three species of animals infected orally with E. granulosus were divided into two groups, PTBA-treated and non-treated control groups. PTBA promoted/enhanced the recovery rate of the worms until 5 days, but none or only a few worms were found in PTBA treated animals thereafter. The highest recovery rate was obtained from M. meridianus treated with PTBA on day 5 post-infection. Some worm developments were observed on days 5 and 10 post-infection. These results demonstrate that M. meridianus could be useful as an alternative definitive host of Echinococcus. PMID- 11871150 TI - Expression of lipoprotein receptors in the aortic walls of diabetic APA hamsters. AB - Syrian hamsters of the APA strain (APA hamsters) have recently been demonstrated to develop atheromatous lesions in the aortic arches under the diabetic condition induced by a single injection of streptozotocin (SZ). Various lipoprotein receptors are reported to play important roles in atherogenesis mainly in vitro, while there are few reports on the relative expressions of these receptors in vivo. In this study, we therefore examined messenger RNA (mRNA) expressions of several lipoprotein receptors on the aortic arches of diabetic APA hamsters at 6, 14 and 26 weeks after the injection (WAI) of SZ. In semi-quantitative RT-PCR, scavenger receptor (SR)-AI, macrosialin (MS)/CD68, and receptor for advanced glycation end-products (RAGE) mRNAs showed significant increases at 6 WAI of SZ, and SR-AI and CD36 mRNA obviously increased until 26 WAI, as compared with the control. Low-density lipoprotein receptor mRNA showed a significant decrease at 14 and 26 WAI, and SR-BI mRNA significantly decreased at 6 and 14 WAI, as compared with the control. Very low-density lipoprotein receptor mRNA was at the same level as the control. By means of in situ hybridization, SR-AI, MS/CD68 and RAGE mRNA were detected in the foam cells of the fatty streaks at 6 WAI, which suggested that SR-AI, MS/CD68 and RAGE play crucial roles in the formation of the fatty streaks, the initial lesions of atherogenesis in diabetic APA hamsters. SR AI and CD36 were also believed to be related to the progression of atherogenesis in this model. PMID- 11871151 TI - Hepatic iron accumulation is not directly associated with induction of DNA strand breaks in the liver cells of Long-Evans Cinnamon (LEC) rats. AB - Effects of accumulation of copper and iron on induction of DNA strand breaks were investigated in Long-Evans Cinnamon (LEC) rats that spontaneously develop fulminant hepatitis. Copper and iron accumulated in the liver of LEC rats in an age-dependent manner from 4 to 15 weeks. Low-iron diet prevented the accumulation of iron in the liver, but did not prevent accumulation of copper. The amounts of DNA strand breaks that were estimated by comet assay in the liver cells of rats fed standard diet increased with age from 4 to 15 weeks. No significant differences were observed in the proportions of LEC rat liver cells without tail and the average lengths of tail momentum in the comet images between LEC rats that had been fed standard MF diet and low-iron diet. These results support the idea that accumulation of iron is not directly associated with the induction of DNA damage in the liver cells of LEC rats. PMID- 11871152 TI - Diurnal variation of heart rate, locomotor activity, and body temperature in interleukin-1 alpha/beta doubly deficient mice. AB - This study was investigated the roles of interleukin-1 (IL-1) on diurnal rhythms of heart rate (HR), locomotor activity (LA), and body temperature (BT). For this purpose, HR, LA, and BT were recorded from conscious and unrestrained IL-1 alpha/beta doubly deficient (KO) and normal C57BL/6J mice using a telemetry system. These parameters were continuously recorded from just after to 2 weeks after transmitter implantation, because we thought that the surgical stress induced IL-1 might affect the biobehavioral activities of the animals. At 1 day after implantation, HR and LA in IL-1 alpha/beta KO mice were higher than those in C57BL/6J mice. While BT in IL-1 alpha/beta KO mice was lower than that in C57BL/6J mice. Moreover, diurnal rhythmicity in these parameters after implantation in IL-1 alpha/beta KO mice appeared earlier than in C57BL/6J mice. At 2 weeks after implantation, there were no significant differences in the light and dark-phase values of each parameter between IL-1 alpha/beta KO and C57BL/6J mice, however, IL-1 alpha/beta KO mice showed clear ultradian rhythmicity. It is thought that a phenotypical difference in biobehavioral activities between IL-1 alpha/beta KO and C57BL/6J mice may reflect IL-1 induced febrile and behavioral responses. These results suggest that IL-1 may play important physiological and pathophysiological roles on biobehavioral activities. PMID- 11871154 TI - Appendectomy in rabbits with extended unilateral anesthesia. AB - Thoracic paravertebral anesthesia was not believed to accompany numbness in the lumbar nerve region. However, we recently discovered that thoracic paravertebral anesthesia could produce analgesia in the lumbar region. We called this block extended unilateral anesthesia. In this study, appendectomy was attempted in rabbits with extended unilateral anesthesia. After a catheter was inserted into the endothoracic fascia in the paravertebral region on the right side at the level of the 11th thoracic vertebra, a 3-ml dose of 2% mepivacaine was injected repeatedly through the catheter. After an injection of the local anesthetic we could observe motor and sensory paralysis unilaterally from the chest down to the lower limb in all the rabbits, the extended unilateral anesthesia. With this anesthesia, we could accomplish appendectomy. This is the initial report of extended unilateral anesthesia applied to appendectomy in rabbits. We think that this anesthesia could be beneficial in future medical and veterinary use. PMID- 11871155 TI - Effects of maternal-melatonin treatment on open-field behaviors and hypertensive phenotype in spontaneously hypertensive rats' (SHR) offspring. AB - Effects of maternal-melatonin treatment in spontaneously hypertensive rats (SHR) were investigated in their offspring. Pregnant SHR were given drinking water with/without melatonin (20 micrograms melatonin/ml tap water) during pregnancy and the lactation period. Maternal-melatonin treatment did not cause changes in body weights during 7 to 27 weeks. Melatonin administration up to weaning period via mother caused a decrease in systolic blood pressure (BP) during 11 to 27 weeks in their offspring compared with those of control group. Open-field behaviors in the offspring were observed at 24 weeks age. Both the control and treatment groups had ratios of central and peripheral locomotion of 30% and 70%, respectively. The treatment group exhibited less total locomotor activity and rearing than the control group did, whereas more latency was exhibited in the treatment group compared with that of the control group. These findings suggest that maternal-melatonin administration may modify open-field behaviors as well as the hypertensive phenotype in their progeny. PMID- 11871153 TI - Diurnal fluctuations of heart rate, body temperature and locomotor activity in the house musk shrew (Suncus murinus). AB - Diurnal fluctuations of heart rate (HR), body temperature (BT) and locomotor activity (LA) in the unanaesthetized and unrestrained house musk shrew (Suncus murinus) were studied using a telemetry system. Six adult male shrews (Jic:SUN) weighing 60-70 g were used in the present study. They were housed under conditions of 24 C and a 12/12-hr light-dark cycle. HR, BT and LA were recorded over 10 days, following the post-implantation period (10 days or more) of the telemetric transmitter. A clear nocturnal rhythm of LA was shown, while intermittent and short-term LA were shown during the light period. The mean HR was 323.5 +/- 8.8 bpm in the light period and 354.3 +/- 5.2 bpm in the dark period, and the fluctuation of HR showed a nocturnal pattern. A nocturnal pattern was also observed in BT fluctuation, and all animals lowered their body temperature from 35-37 C to approximately 30 C or below, mostly during the light period. The fall of body temperature progressed over 2-3 hr, and then rose to the baseline temperature rapidly within approximately 30 min. While the body temperature fell, HR markedly decreased to approximately 100 bpm. These results suggest that the shrew has unique physiological properties in maintaining metabolic balance which are anticipated to be caused by the dramatic alteration of the autonomic nervous function. PMID- 11871156 TI - Life span and renal morphological characterization of the SAMP1//Ka mouse. AB - The senescence-accelerated-mouse prone 1 (SAMP1) is considered to be a model of accelerated senility and it also develops severe kidney damage. The SAMP1//Ka mouse is a specific pathogen free (SPF) subline of SAMP1. The present study examined the life span of the SAMP1//Ka mouse and morphologically investigated the kidneys of this animal at 3, 4, 5, 9, 12, 15, 18 and 24 months of age. Males survived for an average of 25 months and females for 28 months. The median lifespan was 18 months for males and 20 for females. Focal cell infiltration and thickening of the basement membrane in the glomerular capsules or tubules appeared from 4 months of age. At 12 months old, glomerular lesions with expansion of the mesangial matrix and thickening of the basement membrane as well as scar lesions in the outer cortex appeared, and amyloid was deposited in the interstitium or glomeruli from 18 months of age. Morphometrically, although the area of the kidney sections was increased at 24 months of age, the diameter of the renal corpuscles, the number of nuclei of the proximal convoluted tubules and the percentage of renal corpuscles with a cuboidal glomerular capsule did not change with age. The results of the present study indicate that the life span of the SAMP1//Ka is increased and that their age-related renal changes differ from those of the original SAMP1. PMID- 11871157 TI - Postnatal changes in the rheological properties of the aorta in Sprague-Dawley rats. AB - Postnatal changes in the rheological properties of the aortic wall were investigated in relation to morphological development of the wall in Sprague Dawley (SD) rats at 3, 8 and 20 weeks old. The mechanical tensile characteristics of the longitudinal wall strip excised from the proximal thoracic aorta were assessed with stress-strain and stress-relaxation tests. Wall tension in the low and medium strain ranges was significantly lower in 3-week-old rats than in 8 week-old rats and in 8-week-old rats than in 20-week-old rats. Wall stress was significantly lower in 3-week-old rats than in 8- and 20-week-old rats mainly in the medium strain range, but was significantly greater in 3-week-old rats than in 8- and 20-week-old rats in the high strain range. The value of incremental elastic modulus at 3 weeks old was significantly smaller than that at 8 and 20 weeks old at a strain of 0.25 and significantly larger than that at 8 and 20 weeks old at a strain of 0.50. The value of relaxation strength at 5 min after the stretching was significantly greater at 3 weeks old than that at 8 and 20 weeks old. The wall was viscoelastic in the low and medium strain ranges at 3 weeks though large wall stress was generated in the high strain range. Histological investigation revealed that the smooth muscle layer, fine elastin fiber connecting thick elastin fibers and wall thickness were thin at 3 weeks old in comparison with those at 8 and 20 weeks old, though there was no significant difference in number of nuclei of the smooth muscle cells among the three age groups. Changes in the tensile characteristics of the wall reflected well those of the microstructure of the wall with growth. The rheological properties and microstructure of the aortic wall were close to maturation at 8 weeks in SD rats. PMID- 11871158 TI - Functional and histochemical analysis on pancreatic islets of APA hamsters with SZ-induced hyperglycemia and hyperlipidemia. AB - To clarify how Syrian hamsters of the APA strain (APA hamsters) keep a diabetic condition for a long period, the functional and histochemical changes in the pancreatic islets of diabetic APA hamsters were examined. By glucose tolerance test, no glucose-induced insulin secretion was seen in the diabetic APA hamsters. By immunohistochemistry, it was revealed that at 24 hr after SZ-injection, the number of islets had decreased and that remnant islets had become markedly smaller. The islets had hardly any insulin-immunoreactive cells and consisted of cells stained by anti-glucagon and somatostatin antibodies. One, three and six months after SZ-injection, a small number of cells with vacuolative changes, which were positive for PAS staining, were observed in most islets and the vacuolated cells were stained mainly by anti-insulin antibody. In addition, a number of PCNA-positive cells were observed, especially in the periphery of the vacuolated cells, while TUNEL-positive cells were not detected. This data suggests that beta-cells proliferating as a result of the replication of the resident beta-cells in islets had fallen into degeneration and necrosis by a stress, such as the glycogen deposition in hyperglycemia and hyperlipidemia. Consequently, secretion of insulin was maintained at low levels, which allowed the hamsters to live without insulin therapy in the diabetic condition for over 6 months. PMID- 11871159 TI - Ranges of diurnal variation and the pattern of body temperature, blood pressure and heart rate in laboratory beagle dogs. AB - Ranges in diurnal variation and the patterns of body temperature (T), blood pressure (BP), heart rate (HR) and locomotor activity (LA) in 61 laboratory beagle dogs were analyzed using a telemetry system. Body temperature, BP, HR and LA increased remarkably at feeding time. Locomotor activity increased sporadically during the other periods. Body temperature was maintained at the higher value after feeding but had decreased by 0.2 C by early the next morning. Blood pressure fell to a lower value after feeding but had increased by 2.8% by early the next morning. Heart rate decreased progressively after feeding and was 14.5% lower the next morning. This study determined that in laboratory beagles the ranges of diurnal variation and patterns of T, BP and HR are significantly different from those reported in humans and rodents, and that over 24 hr these physiological changes were associated with their sporadic wake-sleep cycles of the dogs. PMID- 11871160 TI - Detection of Corynebacterium kutscheri from the oral cavity of rats. AB - A simple and useful method for the detection of C. kutscheri from the oral cavity of living rats was devised. In 10 sacrificed rats from two naturally and subclinically infected conventional colonies, 10(4.28) or 10(3.84) CFU/ml C. kutscheri were isolated from upper incisor swab extractions, while 10(1.38) or 10(1.58) and < 10 or 10(1.56) CFU/ml from the upper soft palate and pharynx, respectively. In another survey with 26 living animals, which were reared on the same rack, organisms were detected from the upper incisor and gingival swabs in 15 of 26 rats (57.7%). The results were reproducible at a second survey 10 days later. No organisms were isolated from any sites of the orally negative rats. These results indicated that culture of swab specimens from the upper incisors and gingivae of incisors is useful for the detection of C. kutscheri infection in living rats. PMID- 11871162 TI - [Many ethical dilemmas of the general practitioner-patient meeting. How to make limits without rejections?]. PMID- 11871161 TI - [Changed routines in connection with first prescription of oral contraceptives. Lower risk of venous thromboembolism with levonorgestrel preparations]. PMID- 11871163 TI - [General practitioners surveyed on ethical conflicts. How do you handle the "reluctant" patient? How do you handle the patient demanding unnecessary treatment?]. AB - Conflicts between the patient's right to make his own decisions and the physician's obligation to promote health may cause ethical problems in general practice. A questionnaire was sent to 120 Swedish general practitioners; 82 responded, 47 of whom were later interviewed. Faced with a patient reluctant to consent to a medically well founded intervention a majority of the GPs said that they would try to persuade the patient, while they would not agree to medically unjustifiable treatment to patients demanding this. Nor only ethical guidelines, but also a situational ideal of covenant in the patient-physician relationship is desirable in the GP's everyday practice. PMID- 11871164 TI - [Physician's role in grief and loss. Practical guidelines to clinicians in an article in the Annals of International Medicine]. PMID- 11871165 TI - [Carotid endarterectomy in the elderly. Risk of ischemic stroke is reduced in patients older than 75 years]. AB - Epidemiological studies indicate that the risk of first ever stroke is strongly related to age. Based on the NASCET and ECST results, carotid endarterectomy (CEA) is a well-established indication for the prevention of ischemic stroke in patients with 70-99% symptomatic internal carotid artery (ICA) stenosis and in selective cases with moderate stenosis (50-69%). A recent analysis of data from the NASCET group, clearly demonstrates that elderly patients (age > 75 years) with symptomatic ICA stenosis benefit more from CEA compared to younger patients. However, the NASCET collaborators point out that the greater benefits achieved in patients older than 75 years compared to other age groups in the trial depends on the fact that CEA was performed on the elderly having a higher risk of ipsilateral ischemic stroke if treated medically and the lowest surgical risk. In order to obtain the treatment benefit reported, operation performed by skilled surgeons and meticulous medical screening of the elderly patients is recommended to exclude life-threatening conditions. PMID- 11871166 TI - [Children and adolescents with Down syndrome. Continuous ophthalmological monitoring crucial!]. AB - Children with Down's syndrome are at increased risk of ocular manifestations, which is confirmed by a study in Uppsala, Sweden. Sixty percent of the children were wearing glasses and/or had other ocular problems. However, lapses were found in follow-up and referral to the eye clinic. Good visual function is an important prerequisite for optimal quality of life of children and adults with Down's syndrome. Since this group does not always spontaneously get in touch with the health care system including ophthalmologists, the infant must be referred by the pediatrician immediately after birth. The number of individuals with Down's syndrome in Sweden is large (3,500-5,000), and an ophthalmological follow-up program, as presented in this paper, is of key importance. PMID- 11871168 TI - [Increased risk of pregnancy complications and fetal death among obese women]. PMID- 11871167 TI - [Pregnancy and work environment. Practical guidelines for risk assessment]. AB - Pregnant women are a natural part of the workforce in Sweden. A few types of exposure in the work environment are strictly regulated for pregnant women due to well-known risks of miscarriage, malformations or mental retardation in the child. However, a much larger number of types of exposure may pose risks for negative effects on the pregnancy. Chemical, physical and psychosocial factors may interact with personal medical conditions, and a risk assessment must always be performed with the individual in mind. This article addresses the most common questions about pregnancy and the work environment. According to Swedish law, an employer must perform an assessment of the risks in the work environment for the pregnant woman. If the work environment is considered to be hazardous, the particular exposure in question should be reduced or work tasks changed. If neither of these are possible, the woman has a right to compensation from the Social Insurance Office. Self-employed women are currently excluded from this right. PMID- 11871169 TI - [Not all women use maternal health services. Language barriers and fear of the examination are common]. AB - A study within a multinational collaborative study (Barriers and Incentives to Prenatal Care in Europe) showed that out of 3,278 women in Sweden who delivered during a 12-month study period, 5% had registered late (after 15 gestational weeks) at the prenatal care centre or had visited the centre less than three times. Compared to the control group, these women were more likely to be young, single, multipara, immigrant, not fully employed, or having an unplanned pregnancy. The results indicate that their contact with prenatal services could be facilitated by access to interpreters and by transcultural competence among centre staff. PMID- 11871170 TI - [Can women manage without so many check-ups during pregnancy?]. PMID- 11871171 TI - [Ethics in practice--a question of correct prioritization. The decision rests on the individual after all]. PMID- 11871172 TI - [Less speed when it comes to new therapeutic methods!]. PMID- 11871173 TI - [Frustration at the Backen's health center in Umea: "The most needy must wait"]. PMID- 11871174 TI - [In Ostergotland we work "vertically" and "horizontally"]. PMID- 11871175 TI - [The county council of Vasterbotten: distribution of responsibility between politicians and health personnel]. PMID- 11871176 TI - [How do we manage our economic assignments?]. PMID- 11871177 TI - [The good and the evil dragon--myths in the art and medicine]. PMID- 11871178 TI - [William Gilbert--physicist, scientist and physician]. PMID- 11871179 TI - [Misleading report concerning our work on mammographic screening]. PMID- 11871180 TI - [Specialist education in psychiatry must be changed. Stop the compulsory psychotherapy training!]. PMID- 11871181 TI - [Widows should exercise--for the health!]. PMID- 11871183 TI - [A speech in the St Jacob's church on the World AIDS Day 1 December 2001]. PMID- 11871182 TI - [Angiotensin 2 receptor blockaders: how do we explain the kidney-protective effect?]. PMID- 11871184 TI - [Vaccination against HPV infections is future strategy against cervix cancer]. PMID- 11871185 TI - [The law against female circumcision a paper tiger?]. PMID- 11871186 TI - [Richard Horton, The Lancet's editor-in-chief. "We ask questions about medicine no one else dare to ask"]. PMID- 11871187 TI - [Xenotransplantation is the hope of the future. Genetic modification of the donor, the pig, is better than fighting organ rejection by submitting patients to immunosuppression with frequent side effect]. PMID- 11871188 TI - [When does high pressure become a disease? American studies recommend treatment of moderately increased blood pressure--but is increasing the sale of antihypertensive agents the true reason?]. PMID- 11871189 TI - [Placebo--something to believe in?]. PMID- 11871190 TI - [Dose-effect relation of inhalation steroids in asthma should be paid attention to. Comment to a Cochrane report]. AB - The Cochrane collaboration has performed a meta-analysis of all studies found on the dose-effect relation with beclomethasone dipropionate (BDP) in the treatment of asthma. Fifteen studies were found and included, of which only two comprised children. Daily doses of 100 to 2000 micrograms were used. A two-fold increased dose was used in 9 studies but only few significant improvements were seen with the higher dose. A four- to fivefold increased dose gave significant improvement in one of two studies with an improvement of FEV1 of 16% versus 7% with the lower dose. The dose-effect of BDP seems to be very flat and a four-fold increase of the dose seems more reasonable than the two-fold commonly used today, when improved effect is desired. This increases the risk of side effects, and for long term therapy other alternatives should be considered. PMID- 11871191 TI - [Type of primary headache can be diagnosed with the help of careful anamnesis. An account of and comments to a comprehensive review article]. PMID- 11871192 TI - [Circulatory failure in acute poisoning--new therapeutic guidelines]. PMID- 11871193 TI - [Stereotactic radiosurgery of congenital cerebral vessel abnormalities]. PMID- 11871194 TI - [BNCT--neutron radiotherapy of brain tumors]. PMID- 11871195 TI - [The inventor John Ericsson was stricken down by Bright disease]. PMID- 11871196 TI - [Insufficient health information on Internet. Physicians are still necessary as intermediaries and interpreters of the information]. PMID- 11871197 TI - [Three books on one CD-ROM. ICDplus--software for searching classification codes]. PMID- 11871199 TI - [A bio-psycho-social model doesn't explain whiplash injury]. PMID- 11871198 TI - [Surgical emergency services must be changed to manage recruitment of new personnel]. PMID- 11871200 TI - [Mammographic screening is not evidence-based]. PMID- 11871201 TI - [Justified criticism of insufficient confidentiality in connection with paragraph 7 examination]. PMID- 11871202 TI - [Is professional cooperation a bad thing?]. PMID- 11871203 TI - [The dialogue is necessary! Autonomy when it comes to Jehovah's witnesses and blood transfusion]. PMID- 11871204 TI - [How can drug costs be reduced]. PMID- 11871206 TI - [Final reply: AGA destroys a well-functioning system for NO therapy]. PMID- 11871207 TI - [Treatment of deep venous thrombosis]. PMID- 11871205 TI - [Medical snake with many explanations]. PMID- 11871208 TI - [Interventions to improve the management of diabetes mellitus in primary health care and outpatient community settings]. AB - This review should be cited as: Renders CM, Valk GD, Griffin S. Wagner EH, Eijk JThM van, Assendelft WJJ. Interventions to improve the management of diabetes mellitus in primary care, outpatient and community settings (Cochrane Review). In: The Cochrane Library, Issue 2, 2001. Oxford: Update Software. A substantive amendment to this systematic review was last made on 29 June 2000. Cochrane reviews are regularly checked and updated if necessary. BACKGROUND: Diabetes is a common chronic disease that is increasingly managed in primary care. Different systems have been proposed to manage diabetes care. OBJECTIVES: To assess the effects of different interventions, targeted at health professionals or the structure in which they deliver care, on the management of patients with diabetes in primary care, outpatient and community settings. SEARCH STRATEGY: We searched the Cochrane Effective Practice and Organisation of Care Group specialised register, the Cochrane Controlled Trials Register (Issue 4 1999), MEDLINE (1966 1999), EMBASE (1980-1999), Cinahl (1982-1999), and reference lists of articles. SELECTION CRITERIA: Randomised trials (RCTs), controlled clinical trials (CCTs), controlled before and after studies (CBAs) and interrupted time series (ITS) analyses of professional, financial and organisational strategies aimed at improving care for people with Type 1 or Type 2 diabetes. The participants were health care professionals, including physicians, nurses and pharmacists. The outcomes included objectively measured health professional performance or patient outcomes, and self-report measures with known validity and reliability. DATA COLLECTION AND ANALYSIS: Two reviewers independently extracted data and assessed study quality. MAIN RESULTS: Forty-one studies were included involving more than 200 practices and 48,000 patients. Twenty-seven studies were RCTs, 12 were CBAs, and two were ITS. The studies were heterogeneous in terms of interventions, participants, settings and outcomes. The methodological quality of the studies was often poor. In all studies the intervention strategy was multifaceted. In 12 studies the interventions were targeted at health professionals, in nine they were targeted at the organization of care, and 20 studies targeted both. In 15 studies patient education was added to the professional and organisational interventions. A combination of professional interventions improved process outcomes. The effect on patient outcomes remained less clear as these were rarely assessed. Arrangements for follow-up (organisational intervention) also showed a favourable effect on process outcomes. Multiple interventions in which patient education was added or in which the role of the nurse was enhanced also reported favourable effects on patients' health outcomes. REVIEWERS' CONCLUSION: Multifaceted professional interventions can enhance the performance of health professionals in managing patients with diabetes. Organisational interventions that improve regular prompted recall and review of patients (central computerised tracking systems or nurses who regularly contact the patient) can also improve diabetes management. The addition of patient-oriented interventions can lead to improved patient health outcomes. Nurses can play an important role in patient oriented interventions, through patient education or facilitating adherence to treatment. PMID- 11871209 TI - [Laboratory diagnostics in rheumatology]. AB - Peripheral blood cytology and acute phase reactants are used to distinguish between non-inflammatory and inflammatory rheumatic diseases, whereas chronic phase reactants and immunoglobulins can give guidance in distinguishing acute from chronic inflammatory conditions. The complicated path to a well-defined diagnosis of rheumatic disease involves autoimmune serology. This review puts special emphasis on the rational use of autoantibody results for diagnosis, prognosis, planning of follow-up, and estimating the need for therapy. A clinically important use of serodiagnostics can only be implemented by a close collaboration between clinicians and laboratory specialists. A tentative diagnosis is the best basis for ordering and interpreting laboratory results. PMID- 11871210 TI - [Is heart rate reduction beneficial to patients with heart insufficiency? Focus on beta-blockade]. AB - The aim of this review was to assess the effect of pharmacological reduction of the heart rate on prognosis in patients with chronic heart failure. Although a high heart rate is associated with a reduced life expectancy and causes a number of unwanted pathophysiological effects on the failing heart, there is no documentation that the beneficial effect of beta-blockers or other heart failure agents on survival is mediated through a reduction in heart rate. PMID- 11871211 TI - [Supervision]. AB - Within the past year, the idea of supervision has been introduced and promoted in the media by the manufacturers of wavefront-guided excimer lasers for refractive surgery. The concept is related to the ability of obtaining supernormal visual acuity and contrast sensitivity by correction of higher order aberrations in the biological optics of the human eye. However, the ocular aberrations fluctuate continuously and dynamically over time with the functional state of several of the individual optical components, including: thickness of the tear film, size of the pupil, and the degree of lens accommodation. These temporal variations represent major obstacles for achieving aberration-free images. In addition, the central perception of ideal aberration-free images may be hindered by neuronal limitations in the retina and brain. So far, only a few patients have experienced an increase in best corrected visual acuity following wavefront-guided refractive surgery. Also in the first prospective clinical trials with myopic individuals, only modest differences in visual performance have been detected between wavefront-guided and conventional treatment. Thus, immediate expectations from the new technology should be tuned down; supervision is not waiting around the corner. PMID- 11871212 TI - [Local thrombolysis in proximal deep venous thrombosis of the lower extremity]. AB - INTRODUCTION: Deep venous thrombosis (DVT) often leads to chronic venous insufficiency and the present study was conducted in order to investigate the effectiveness of catheter-directed thrombolysis in patients with proximal DVT of the lower extremity (iliac vein involved), with respect to recanalisation and maintenance of venous valve function. MATERIAL AND METHODS: A prospective clinical investigation was carried out with puncture of the popliteal vein for continuous infusion of r-alteplase. Twelve patients suffering from recent proximal DVT were treated: In 10 patients the left extremity was affected, in two the right. Three of the 12 patients had factor V Leiden mutation in the heterozygote form, one of whom also had prothrombin mutation in heterozygote form. RESULTS: Ten of the 12 had their venous thrombosis successfully lysed and were discharged with an open venous system in the affected limb. The lysed venous segments remained patent in all ten, with normal venous valve function, as evaluated by Doppler reflux testing. The median follow-up time was five months (range 0-9 months). In one patient, the proximal thrombus (iliac) was lysed successfully, but the femoral vein could not be opened, probably because of an old thrombus remaining from a previous DVT episode. In the other patient, the venous thrombus was lysed successfully, but the vein rethrombosed after one day. DISCUSSION: Catheter-directed thrombolysis appears feasible in patients with recent proximal DVT and the short-term results are good in terms of venous patency and valve function. A randomised trial is necessary to test whether this treatment modality is superior to conventional anticoagulation therapy. PMID- 11871213 TI - [The CARMEN trial: increased intake of carbohydrates--simple or complex--and unchanged blood lipids in overweight subjects]. AB - INTRODUCTION: The purpose was to investigate the long-term effect of ad libitum low-fat, high-carbohydrate diets with mainly simple or complex carbohydrates on body weight and blood lipids. MATERIAL AND METHODS: A six-months controlled randomised multicentre trial with the participation of five European centres. A total of 398 moderately obese subjects (BMI: 30.4 +/- 2.7 kg/m2, mean +/- SD) were randomised to four groups: Control diet group with normal macronutrient intake, a low-fat/high simple carbohydrate group, a low-fat/high complex carbohydrate group and a seasonal control group. RESULTS: After six months we found a drop in body weight of 1.7 kg (p < 0.05) in the simple and of 2.6 kg (p < 0.001) in the complex carbohydrate group compared with the control diet group. Fat mass decreased by 1.9 kg (p < 0.05) and 2.4 kg (p < 0.001) in the simple and complex carbohydrate group, respectively, compared with the control diet group. There were no significant changes or group differences in fasting blood lipids, glucose, insulin, or leptin. DISCUSSION: The results show that it is favourable to replace dietary fat by carbohydrates (simple or complex) in relation to body weight regulation. No detrimental effects were seen on blood lipids in contrast to previous contentions. Our results underline the importance of a low-fat/high carbohydrate diet in the management of obesity and the ensuing health problems. PMID- 11871214 TI - [Transcatheter closure of atrial septal defects]. AB - INTRODUCTION: Transcatheter based device closure of secundum atrial septal defects (ASD) have been offered at our institution since September 1997. During the first three years, closure was attempted in 56 patients aged 3 to 71 years (median 15). MATERIAL AND METHODS: Hospital notes on all patients undergoing device closure of atrial septal defects between September 1997 and September 2000 were reviewed. Indications for attempted closure were significant arteriovenous shunting (n = 49), venoarterial shunting (n = 3), and suspected paradoxical embolism (n = 4). RESULTS: Device delivery was achieved in 51 (91%) patients. In five patients, the procedure was abandoned, owing to the unsatisfactory position of the device. In one patient, embolisation occurred within 24 hours. At three months follow-up, 44 of the 46 patients investigated had completely closed defects. Residual flow was seen in two patients, including one (with three defects) who subsequently underwent elective surgical closure. There were no other complications. DISCUSSION: Device closure of ASD is a realistic alternative to surgical treatment. PMID- 11871215 TI - [Surgical treatment of preauricular fistula. A follow-up study]. AB - INTRODUCTION: Our aim was to evaluate the results of surgery on preauricular fistulas over a ten-year period and to determine the postoperative recurrence rate and the factors responsible. MATERIALS AND METHODS: Forty-three patients with 51 preauricular fistulas, who had been operated on in the period, 1 August 1990 to 1 August 2000, at the Department of Otolaryngology, Aarhus University Hospital, were asked to participate in a follow-up examination. Thirty-three patients turned up for examination, four patients were interviewed over the telephone, and for the rest the data available from the medical records were used. RESULTS: Surgical extirpation was performed on 51 fistulas. Eight fistulas had previously been operated on. The recurrence rate of the fistula primarily operated on at our department was 5%. The recurrence rate of secondary surgery was 63%. Five fistulas were infected at the time of surgery, three recurred. Seven patients developed a postoperative abscess, six of whom had a recurrence. All the fistulas were blind-ended. In 94% of the histology tests, the fistula was lined with multilayered squamous epithelium. Sixteen patients had a family history of the condition. CONCLUSION: Preauricular fistula is a benign disease that often stays asymptomatic, but can become troublesome if infected. The treatment is surgical extirpation. The recurrence rate rises if the fistulas are infected at the time at surgery or if there has been an earlier attempt of extirpation. This emphasises the importance of an extended knowledge of the disease, so that patients can be referred to an ENT specialist should symptoms occur. PMID- 11871217 TI - [Xanthogranulomatous pyelonephritis]. AB - Xanthogranulomatous pyelonephritis is a rare disease probably caused by long standing obstruction and secondary infection. We report a case in a 47-year-old woman with no previous history of renal stones. Preoperative diagnostic workout included ultrasonography, CT scan, and drainage. Nephrectomy was performed, and the patient's postoperative recovery was uneventful. The difficulties of correct preoperative diagnosis are discussed. PMID- 11871216 TI - [Lithium induced dysfunction of the parathyroid hormone]. AB - The prevalence of hyperparathyroidism (HPT) in patients treated with lithium is higher than that in controls. Lithium seems to affect calcium metabolism, by acting directly parathyroid hormone cells, and distal tubuli in the kidneys. Because hypercalcaemic HPT can cause psychiatric symptoms mistakenly attributed to the lithium treatment, ionised calcium should be a standard control. PMID- 11871218 TI - [Prognostic value of PET and MRS in the assessment of cerebral status of children]. AB - We report a case of a 12-week-old previously normal infant with severe brain damage after an episode of asphyxia during an RS-virus infection. Sub-acute MRI was normal, but new functional techniques, PET (positron emission tomography) and MRS (magnetic resonance spectroscopy) were severely abnormal. At an outpatient clinic three months later, he had developed microencephaly and the MRI was now severely abnormal. The case shows the importance of using multimodality functional imaging techniques to assess the cerebral status of infants for prognosis and course of treatment. PMID- 11871219 TI - [Intraoperative burns. An unforeseen danger?]. AB - Two patients were admitted to the department of burns in Copenhagen. Both had suffered burns following uncontrolled ignition of chlorhexidine-alcohol during surgery. The aim of this article is to highlight this potentially detrimental risk, which exists in almost all operating rooms, unless simple precautions are taken. PMID- 11871220 TI - [Early goal-directed life-saving therapy of sepsis]. PMID- 11871221 TI - [The Danish Medical Society wishes the concept of specialty be based on the scientific development]. PMID- 11871222 TI - [Big nutrition surveys are effective!]. PMID- 11871223 TI - [Criticism of the Cochrane review on mammographic screening is not justified]. PMID- 11871224 TI - [Does paracetamol increase the risk of gastric hemorrhage and perforation?]. PMID- 11871225 TI - [Paracetamol increases the risk of upper gastrointestinal hemorrhage and perforation]. PMID- 11871226 TI - [Danish biomedical research in the postgenomic era--who will take the initiative?]. PMID- 11871227 TI - [Histochemical and morphometric evidence of the participation of dorsal hippocampal neurons in the antistress effect of melatonin]. AB - The epiphyseal hormone melatonin (0.1 mg/kg) does not influence the functional state of pyramidal neurons in the dorsal hippocamp of intact rats, but decreases the swimming-stress-induced activation of these neurons. Removal of the pineal gland increased the functional activity of neurons in the intact rats and potentiated the stress-induced histochemical and morphometric changes. PMID- 11871229 TI - [Effect of piracetam on the conditional reflex memory under probable reinforcement conditions]. AB - The effect of piracetam on the development of a two-link system of feed-procuring conditioned reflexes was studied in rats with probability reinforcement. The experiments showed that piracetam improves both the current learning process and the memory track (previous experience) reproduction. In addition, the drug decreases the role of the probabilic character of reinforcement, thus facilitating the synthesis of two reflexes in a united functional complex. PMID- 11871228 TI - [Effect of the new potential anti-Parkinson agent, hymantane, on levels of monoamines and their metabolites in rat striatum (a microdialysis study)]. AB - The new aminoadamantane derivative N-(2-adamantyl)hexamethyleneimine hydrochloride (A-7, hemantane) exhibited a pronounced antiparkinsonian effect on various experimental models. Hemantane showed a broad activity spectrum, being superior to amantadine (midantane) in some tests. Administered in an effective antiparkinsonian dose, hemantane increased the extracellular dopamine content in the striatum. The drug also produced a dose-dependent decrease in the extracellular level of dihydroxyphenylacetic acid and homovanillic acid (DOPAC and HVA, the dopamine metabolites) and 5-hydroxyindoloacetic acid (5-HIAc, a serotonin metabolite) in the striatum, which may reflect inhibition of the monoamine oxidase activity in the brain. These results show that the dopaminergic and serotoninergic nigrostriatial systems are involved in the mechanism of hemantane action. PMID- 11871230 TI - [New approaches to analysis of the interrelationship between cholinergic and dopaminergic mediator systems]. AB - Intermediatory relationships between the cholinergic and dopaminergic neurotransmission systems were analyzed using published data and the original experimental results obtained on Daphnia magna Straus, a new test object. Based on these results, the antihaloperidol activity of a series of M- cholinoblocking agents with different receptor selectivities were studied in comparison to the new cholinoblocker pentifin exceeding in the activity the classical antiparkinsonian drugs such as cyclodol, amedin, and norakin. PMID- 11871232 TI - [Stimulation of peripheral delta1-opioid receptors as a method of preventing ischemic and reperfusion arrhythmia: role of K(ATP)-channels]. AB - Preliminary administration of the delta 1-opioid receptor (delta 1-OR) peptide agonist DPDPE (0.5 mg/kg, i.v.) decreased the incidence of occlusion (10 min) and reperfusion (10 min) arrhythmias in rats. The delta 2-OR agonist DSLET did not affect arrhythmias upon the coronary artery occlusion and reperfusion. Pretreatment with the selective delta-antagonists ICI 174,864 (2.5 mg/kg) or TIPP[psi] (0.5 mg/kg) completely eliminated the antiarrhythmic effect of DPDPE. Uncapable of crossing the blood brain barrier, the nonselective OR antagonist naloxone methiodide (5 mg/kg) also abolished this effect. At the same time, hexametonium (10 mg/kg) did not antagonize the antiarrhythmic effect of DPDPE. Pretreatment with the KATP channel blocker glibenclamide (0.3 mg/kg) completely eliminated the protective effect of the delta 1-OR stimulation. It was concluded that the delta 1-OR stimulation prevents the ischemic and reperfusion arrhythmias by means of the KATP channel activation. PMID- 11871231 TI - [Participation of central and peripheral I1-imidazollinovykh and alpha2 adrenergic receptors in realizing the effects of moxonidine]. AB - The role of the central and peripheral I1-imidazoline and alpha 2-adrenergic receptors in the development of hypotension and bradycardia induced by the intravenous injections of moxonidine was studied, in stroke-prone spontaneously hypertensive (SHR-SP) rats. The intravenous and local injections of I1--alpha 2 antagonist efaroxan (Efa) into the rostral ventrolateral medula (RVLM) abolished the moxonidine-induced fall of blood pressure, but did not eliminate bradycardia. The alpha 2 antagonist yohimbine (Yoh) injected into RVLM did not influence the moxonidine effects, whereas the intravenous Yoh injections fully blocked these effects. Thus the hypotensive action of moxonidine is mediated predominantly by the I1-imidazoline RVMM receptors, while the heart rate decrease is partly realized via the alpha 2 adrenoreceptors of peripheral innervation. PMID- 11871234 TI - [Cardioprotective effect of drugs with antioxidant activity in acute cerebral ischemia]. AB - The bioelectric cardiac activity was studied in the experiments on white mice with an acute cerebral blood circulation disorder. It was found that he resulting EEG changes possess a specific character, with the sympathoadrenal system stimulation playing an important role in the acute cerebrocardiac syndrome development. The antioxidant-type agents such as emoxypine (50 mg/kg), mexidol (50 mg/kg), and cytochrome C (10 mg/kg) produce a significant cardioprotective effect in the test animals with experimental cerebral ischemia, which was comparable with the effect of propranolol (obsidane) (0.1 mg/kg). PMID- 11871233 TI - [Effect of nootropic agents on impulse activity of cerebral cortex neurons]. AB - The effect of nootropes (semax, mexidol, and GVS-111) on the activity of individual neurons in various cerebral cortex regions was studied by microelectrode and microionophoresis techniques in cats immobilized by myorelaxants. It was established that the inhibiting effect of mexidol upon neurons in more than half of cases is prevented or significantly decreased by the GABA antagonists bicuculline and picrotoxin. The inhibiting effect of semax and GVS-111 upon neurons in more than half of cases is related to stimulation of the M-choline and NMDA receptors, respectively. PMID- 11871235 TI - [Tolerance to cardiotoxic effect of strophanthane K in experimental total heart block and electrocardiostimulation]. AB - Tolerance to the cardiotoxic effect of strophanthin K was studied in narcotized dogs with experimental complete heart block (CHB) and ventricular electrostimulation. A threshold to the toxic arrhythmogenic effect of strophanthin K was significantly reduced as compared to that in the control group. As the electrocardiostimulation (ECS) frequency increased, the minimum arrhythmogenic dose of strophanthin K showed a growth, while not reaching a level for the sinus rhythm. It is concluded that the basic mechanisms of the arrhythmogenic effect of strophanthin K under the conditions studied are the trigger activity in dogs with ECS and the increased peacemaker's activity in dogs with CHB. PMID- 11871236 TI - [Cerebrovascular and renal effects of cerebrolysin and dependence on salt intake]. AB - Experiments on rats with occluded common carotid arteries showed that an excess sodium chloride consumption increased the loss of test animals as a result of the maximum decrease in the local cerebral blood flow and sharply pronounced brain swelling. The sodium chloride substitute hyposol (giposol) reduced the extent of cerebral ischemia and brain swelling effect and increased the renal perfusion and diuresis levels. In the test animals receiving a high-Na diet, the efficacy of cerebrolysine was less pronounced. In contrast, hyposol increased the antiischemic, saluretic and antiswelling effects of cerebrolysine under the carotid artery occlusion. PMID- 11871237 TI - [Cycloferon in treating duodenal ulcers in rats]. AB - The possibility of using cycloferon (interferon inductor) for a complex treatment (in combination with the main drug solcoseryl possessing pronounced therapeutic properties) of duodenum ulcers was experimentally studied in male rats. The experiments showed a considerable difference in the interferon status of animals with model duodenum ulcers treated with cycloferon, solcoseryl, their combination, and placebo (control). The healing effect of solcoseryl administered in combination with cycloferon exceeded that of each component administered separately. PMID- 11871238 TI - [Effect of Rhaponticum carthamoides extract on hemorheological properties of blood in rats with arterial hypertension]. AB - The hemorheological activity of a dry extract from Rhaponticum (Leuzea) carthamoides (Willd.) Iljin. was studied in spontaneously hypertensive rats (SHRs). A 14-day treatment of the test rats with a daily dose of 150 mg/kg improved the rheological characteristics, as manifested by a reduced viscosity of the whole blood and plasma, increased spontaneous erythrocyte aggregation and fibrinogen concentration, and increased erythrocyte deformability and electrophoretic mobility against the untreated control level. PMID- 11871239 TI - [Effect of quinine and quinidine on the status of hematopoietic and lymphoid organs]. AB - The effect of quinine and quinidine introduction in subtoxic doses on the characteristics of the hemopoietic and lymphoid organs was studied in BALB/c mice. The treatment led to reversible anemia and neutrophile leukocytosis in peripheral blood. There was a tendency to the early decrease in neutrophiles the late decrease in lymphocytes in the bone marrow. The quinine-induced decrease in erythrocytes was accompanied by reticulosis, lowered erythrocyte osmoresistance, and intensified spleen erythropoiesis; quinidine induced reticulocytopenia and a decrease in the bone narrow erythroblasts. The use of both compounds was accompanied by a decrease in thymus weight and increase in the thymocyte loss in the early period, followed by the spleen weight increase in the long term. PMID- 11871240 TI - [Menadione and roboxin as inducers of immunomodulating activity of erythrocyte stroma]. AB - The erythrocyte stroma of intact rats treated with riboxin can be used, after an additional extracorporal treatment with menadione, as a transaphthogenic softly acting immunostimulant. Introduced into rats with a secondary experimental immunodeficient syndrome caused by indomethacin or gentamycin, this stroma leads to normalization of the humoral immune response to ram erythrocytes. PMID- 11871241 TI - [Effect of plant melanin pigment on the clastogenic effects of chemical mutagens in mice]. AB - The chromosome aberration assay in the bone marrow cells of C57BL/6 mice showed that melanin pigment (MP) in a dose range from 0.01 to 10 mg/kg does not influence the clastogenic effect of dioxidine (200 mg/kg, i.p.), while reducing the clastogenic effect of cyclophosphamide (20 mg/kg, i.p.) by a factor of 1.5-4 in various treatment regimes depending on the mutagen injection time. PMID- 11871242 TI - [Seasonal features of the effect of adaptogens on sex behavior of experimental animals]. AB - Seasonal variation of the aphrodiatical activity of pantocrin (deer antler extract), ginseng root tincture, and eleuterococcus extract was studied. It was found that a one-month treatment with these adaptogen preparations in a daily dose of 100 mg/kg (for dry parent substance) stimulate the sexual behavior of puberal mice and rat males in winter. The maximum effect was observed for pantocrin. In summer months, the stimulating effect of adaptogens was not manifested. The reference drug (tentex forte) increased the sexual behavior irrespective of the season, the effect being more pronounced in summer months. These results are explained by assuming that the adaptogens studied possess a weak gonadotropic activity, which favors normalization of the testosterone level (reduced in winter season) in experimental animals. PMID- 11871243 TI - [Electronic counters for recording indicators in biomedical experiments]. AB - The design is described and circuit diagrams are presented for PC-controlled electronic data counters based on microcalculators and integrated circuits. The counters are intended for data accumulation in the course of biomedical experiments. PMID- 11871247 TI - HIV campaign in an Islamic school. PMID- 11871246 TI - The genetic edge. PMID- 11871245 TI - [Effect of methysergide on behavior of rats with various levels of corticosteroids]. AB - The effects of methysergide (metisergid), a nonspecific antagonist of type I and II serotonin receptors, on the character of the conditional reflex, learning and behavior of male rats was studied under conditions of low versus high content of corticosteroids. It was established that methysergide improved the ability of animals to acquire and reproduce the active avoidance reflex in the test animals with a deficit of the corticosteroid hormones, while no such effects were observed in rats with increased level of these hormones. Under conditions of the corticosteroid hormone deficit, methysergide stimulated the total locomotor activity in the open field test; in the excess of corticosteroids, the drug inhibited the emotional and grooming behavior components. PMID- 11871244 TI - [Role of GABA-ergic system of the brain in regulating blood circulation]. PMID- 11871248 TI - SMART starts! PMID- 11871250 TI - P-glycoprotein and HIV. PMID- 11871249 TI - Protein primer. PMID- 11871251 TI - [Multistage mechanism of reproductive sympatric isolation of the closely-related species, Ixodes persulcatus and I. Pavloskyi (Ixodidae)]. AB - The closely related species Ixodes persulcatus and I. pavlovskyi are vectors of agents tick-born encephalitis and Lyme disease. These species have two great disjunctive sympatric distribution areas (fig. 1), where both species inhabit the same biotopes, and during the same season. In these conditions there are prerequisites of contacts of opposite sexes of the different species during ambushing the hosts on vegetation. Sequence of "switching on" of the morphological, physiological, and hostparasite factors of reproductive isolation has been established. The features and the range of manifestation of these factors at different combinations of sexes and species of the partners have been revealed. The studied factors, such as precopulative and postcopulative components of the mechanism of reproductive isolation of I. persulcatus and I. pavlovskyi were considered. A hypothesis for sympatric origin of the species as a result of specialization the adults of I. persulcatus to mammals, and I. pavlovskyi to birds is proposed. PMID- 11871252 TI - [Involvement of the common shrew, Sorex araneus (Insectivora, Soricidae), in circulation of the tick-borne encephalitis virus in south-western Siberia]. AB - We presented the data on the abundance of immature instars of the taiga tick Ixodes persulcatus Schuize on the common shrews Sorex araneus L. in natural foci of tick-borne encephalitis in the south of Western Siberia. Basing on the results of virological and serological studies we demonstrated a low effectiveness of this host species as a donor of disease agent strains, which are predominant in the territory under study, for ticks feeding on shrews. The analysis of samples taken from the young shrews in winter and spring using reverse RNA transcription with polymerase chain reaction and ELISA revealed occurRence of subvirion components of the tick-borne encephalitis (RNA and capsid protein E) ether in brain, liver or spleen in 90 percent of shrews (n = 42). Neither hemagglutination antigen nor infectious virus have been detected. We discussed a possible epizootic role of the maintenance of non-infectious tick-borne encephalitis virus in overwintering animals. PMID- 11871253 TI - [Functional organization of natural tick-borne encephalitis foci and prognosis of their epidemic manifestation: analysis of one-dimensional temporal series of infection]. AB - Temporal extrapolation prognosis of tick-borne encephalitis diseases in the Primorye territory and its certain regions is made by means of autoregression models. The results obtained shows good prospects of used methods and models. PMID- 11871254 TI - [Distribution ranges of malarial mosquitoes other than Anopheles maculipennis species (Diptera, Culicidae) in Russia]. AB - Maps and distribution data for six species of the genus Anopheles in Russia are given. PMID- 11871256 TI - [Photoreactions of the littoral trematode Renicola thaidus (Trematoda: Renicolida)]. AB - The behavioral response of Renicola thaidus cercariae (Renicolidae) to the light has been studied. The positive photo reaction was observed during the first hour of the cercariae free-living period (after shedding from the mollusc). Six hour old cercariae did not show any direct photo reaction. Temporal presence of the positive photo reaction promotes a wider spatial distribution of R. thaidus cercariae. This fact may increases a probability of successful finding and infection of the second intermediate host--bivalves such as the blue mussels. Besides, the more random distribution of infection in bivalve host populations is achieved. This circumstance increases a probability of infected bivalve to be eaten by the final hosts--seabirds. PMID- 11871255 TI - [Some peculiarities of the relationships between parasitic copepods and their invertebrate hosts]. AB - According to the rule of academican E. N. Pavlovskiy, any organism of host is an environment of inhabit for a parasite (Pavlovskiy, 1934). It was analysed, which "ecological niche" or microbiotop (= microhabitat) is occupied by this or that species of symbiotic (parasitic) copepods in organisms of different groups invertebrate-hosts. The assumption lying in a basis of the given analysis means that each group of hosts may give to cohabitants only certain variants of microbiotopes independently on the general morphological structure and life mode of hosts. Five types of microbiotops offered by various groups of hosts for symbiotic copepods are designated (Ta[symbol: see text] 2). 1. The body surface of benthic invertebrates as a microbiotope is characterized by conditions being little different (concerning any kind of physical and chemical influences on copepods) from those in external environment on any other substrate. Apparently a trophical dependence plays a determining role in this case. There are certain directions in a development of adaptations, which are characteristic in some extent for all water ectoparasitic crustaceans and have one functional task--to help to an ectoparasite to keep itself on a surface of host body. In the first, the maxillules and maxillipeds significantly are developed, they get a form of large claws, with which the dopepods are strongly attached on a surface of host body and have an opportunity to move on it without a danger to be washed off. In the second, the form of the body undergoes a dorso-ventral expression and expansion of prosome, forms a cephalic shield allowing to the symbiont to press itself tightly to the host body surface and to avoid the loss of host (tab. 2). In occasions, some ectoparasites stimulate the formation of galls in skin tissues of the host, that also provides the parasite with constant conditions, without any threat to lose the host. However, this phenomenon has not a wide distribution and is observed in some groups of crustacean and echinoderm hosts. 2. The narrow tubular cavities in the organism of host either they are a part of external environment (as in channel system of spongia) or a part of internal environment of organism (as channels of blood system or thin parts of a digestive system) have always rigidly limited sizes and form. Characteristics of all parasites occupying this microbiotopes are the strong transformations. They are expressed by the reduction of legs or any other appendages (frequently in a significant degree), loss of segmentation to some extent and in eruciform (or vermiform) form of a body (tab. 2). This microbiotope is occupied by an ectoparasite in one case only (Spongicola uncifer from channel system of spongia) and by endoparasites in all other cases. 3. Large cavities connected with external environment. The formations of various genesis, such as mantle cavity of molluscs, gill cavity and marsupium of crustaceans, bursal cavity of ophiuroids and branchial cavity of ascidians, concern this type of microbiotopes. All of them are characterized by the relative difference from the external environment and rather large volume (in comparison to sizes of copepods), that provides the parasites with a sufficient protection from factors of the external environment and constant source of food such as elements of host body or food's particles brought by the water flow. Morphological changes in inhabitants of the microbiotope have two directions. They practically are absent in the overwhelming majority copepods, living in the mantle of cavity of the lamellibranches. On the other hand, the inhabitants of gill cavity and marsupium of crustaceans, bursal cavities of ophiuroids and branchial cavity of ascidians are characterized by the presence of strong transformations. Usually there are expressed in a loss of segmentation to some extent, reduction of appendages and swelling of body, as in species of the genus Sphaeronella (tab. 2). Changes are also observed in the life cycle: the tendency to reduce stages of development (development of nauplii stage, which takes place under the ovarial cover). In this case the copepodid stages hatch from the ova. 4. The internal cavity of organism of host. This type of microbiotopes in different groups of the hosts is represented in a various degree. We recognise it in a coelome of polychaetes, lacunar system of molluscs, mixocoel of crustaceans, coelome of echinoderms and cavity of body in ascidians. Two basic evolutionary directions are observed in copepods occupying this microbiotope. In the first case, the parasite is not exposed to transformations and keeps the initial plan of structure as in ancestral free-living forms. In the second case the parasites are exposed to strong transformations, they either live directly in cavity's liquid, or are surrounded by a cyst (as in Cucumaricolidae). 5. Microbiotope of the last type is most specific. The simultaneous existence in two environments- external environment (environment of the second order) and internal environment (environment of the first order) leads to the complete loss of ancestral type in a structure and level of organisation. At the same time both morphological and functional division of the parasite body into two parts produces a new formation- the ectosome and endosome. In this case we deals with the phenomenon of mesoparasitism. PMID- 11871257 TI - [Study of fecundity of Bunodera luciopercae (Trematoda: Bunoderidae)]. AB - The seasonal dynamics of the maturation of trematode Bunodera luciopercae was investigated in 1992-1998. The actual fecundity of adult helminths from the ruff, perch, sander and volga pike-perch in the Rybinsk Reservoir was registered in the end of spring. The accumulation process of eggs reflected the host alteration and depended on the food ration, body length and sex of hosts. PMID- 11871258 TI - [Ciliary activity of cells of gill and leg glimmeral epithelium of Unionidae invaded by trematodes of Aspidogaster conchicola and Bucephalus polymorphus]. AB - The paper provides data concerning the influence of the parasitic worms Aspidogaster conchicola and Bucephalus polymorphus on the rate and duration of ciliary beating of gill and leg in 7 species of Unionidae (Unio conus borysthenicus, U. tumidus falcatulus, U. rostratus rostratus, U. limosus graniger, U. pictorum ponderosus, Colletopterum piscinale falcatum, C. ponderosum rumanicum). The high level of infection of molluscs with B. polymorphus oppresses the glimmeral epithelium activity of gill and leg (by 4.2-32.3%). The weak and moderate levels of infection of molluscs with B. polymorphus do not decrease the glimmeral epithelium activity and sometimes even rise it (by 3.4-8.1%). The presence of few A. conchicola (1-3 individuals) in the organism of mollusc does not change the functioning of glimmeral epithelium. PMID- 11871259 TI - [Phospholipid and fatty acid content of the blood of sheep infected with the nematode Dictyocaulus filaria]. AB - The results of analysis of phospholipids (PL) and fatty acid content in the blood of sheep infected with the nematodes Dictyocaulus filaria are displayed. A significant increase of lysophosphatidylcholine and arachidonic acid as well as a decrease of docozagexaenic acid in PL of infected sheep have been recorded. That points out to structural and functional disorders of cellular membranes during the infection. These disorder could be used as a metabolic criterion to estimate the relationships within the host-parasite system examined. PMID- 11871260 TI - [Cryptobia udonellae sp. n. (Kinetoplastidea: Cryptobiida) - parasites of the excretory system of Udonella murmanica (Udonellida)]. AB - A new cryptobiid flagellates, Cryptobia udonellae sp. n., is described from the excretory channels of Udonella murmanica. The body of flagellates is spindle shaped. The flagellar pocket is subapical. Two flagella emerge from the pocket. One flagellum turns anterior and is forward-directed; the other flagellum is directed posterior and close to the ventral cell surface. The ventral groove is well developed. The cytostome opens just anterior to the flagellar pocket. The cytostome leads to the short cytopharynx. In the excretory channel of worms the flagellates C. udonellae sp. n. are attached to microvilli of epithelium or lay free in the lumen. Both flagellates have been studied with TEM. The unusual parasite system which involves organisms of four different phylums of animals has been described for the first time. PMID- 11871261 TI - [Aging and changes in fecundity of the flea Leptopsylla segnis (Siphonaptera: Leptopsyllidae)]. AB - In experiment, the mean diurnal fecundity of Leptopsylla segnis females increased during the first five days since the beginning of feeding on white mice and reached the maximum in the second five-day period (15.7-16.7 eggs per 1 female during a day). In subsequent period of stay on a host, an activity of egg laying gradually decreased. In flea females, which had lived on hosts for 40 days and more this index was 3.7 eggs. During the mean life longevity (23 days), the flea females lay about 260 eggs; the females, which have lived more than 40 days, produce above 400 eggs. PMID- 11871262 TI - [Two new species of myxosporidians from Japanese flounder from Peter the Great Bay (Sea of Japan)]. AB - Two new species of Myxosporidia, Ceratomyxa costata sp. n., and Leptotheca amatea sp. n. from the gall bladder of the Japanese flounder Limanda yokohamae from the Peter the Great Bay are described. PMID- 11871263 TI - [Early history of helminthology in the Russian Academy of Sciences]. AB - Zoological Institute RAS (up to 1930--Zoological Museum) dates back to 1728, when the first Russian museum, Kunstkammer, was opened under the authority of the newly organized Russian Academy of Sciences. Zoological collections of Kunstkammer were greatly enriched by a number of naturalists who worked in Russia since the middle of the 18-th century, P. S. Pallas (1741-1811) and I. T. Koelreuther (1733-1806) were among them. Both had made an essential progress into helminthology. In 1895, Zoological Museum legislatively acquired a statute of the central institution in Russia for zoological research. Investigations of parasitic worms here become permanent. First parasitologists in its staff, A. K. Mordvilko (1867-1938) and N. P. Annenkova-Khlopina (1887-1950), should be mentioned. PMID- 11871265 TI - [A new approach to modeling the diversity dynamics of Phanerozoic marine biota]. AB - Modeling of fossil diversity dynamics is usually done with the help of the models borrowed from the population dynamics theory. However there are principal differences between organisms and taxa, reproduction and divergence, mortality and extinction that make this approach doubtful. Another model is presented here, in which absolute origination rate does not depend on diversity, the ability of new genera to sustain unpredictable environmental changes increases three times abruptly at Cambrian/Ordovician, Permian/Triassic and Cretaceous/Tertiary boundaries. In this model the diversity increases due to accumulation of long lived genera. The computer simulation showed that the model agrees with empirical data by 15 major criteria. The laws of community evolution apparently can explain the general pattern of punctuated equilibrium in the evolution of marine biota. PMID- 11871266 TI - [Genetic-evolutionary basis of symbiosis doctrine]. AB - The author presents the current notion of symbiosis as one of the main adaptation of an organism to changeable environment. Symbiosis is considered as a super organism genetic system within which there are different interactions (including mutualism and antagonism). Genetic integration of symbiotic partners can be realized as cross regulation of their genes, exchange of gene products (proteins, RNA), gene amplification and sometimes gene transfer between organisms. On the phenotypic level these processes result in signal interactions, integration of partner metabolic systems and development of symbiotic organs. Co-evolution is considered as an assemblage of micro- and macroevolution processes basing on pre adaptations and proceeding under influence of different forms of natural selection (individual, frequency-depended and kin selection). Symbiosis can be compared with sexual process since both are the forms of organism integration characterized by different genetic mechanisms and evolutionary consequences. The genome evolution in symbiotic microorganisms can proceed by: 1) simplification of genome in obligate symbiosis (loss of genes that are necessary for independent existence, transfer of some genes to the host organism); 2) complication of genome in facultative symbiosis (increase in genome plasticity, structural and functional differentiation of genome into systems controlling free-living and symbiotic parts of life cycle). Most of symbiotic interactions are correlated to an increase in genetic plasticity of an organism that can lead to evolutionary saltations and origin of new forms of life. PMID- 11871264 TI - [Correlation of space-time ranges in ecological hierarchies of various nature]. AB - Relations between characteristic scales of time and space are analyzed for the hierarchical systems of a various nature. The available data fit well to the power relation: [T] = a[L]b, where [T] and [L] are characteristic ranges of time and space, b--scaling exponent, a--conversion coefficient. The spatio-temporal scales of ocean physical and biological (pelagic) processes are closely overlapped. Contrastingly, the scale for terrestrial and benthic ecosystems and their environments differ noticeably. For terrestrial and benthic ecological systems, and also for atmospheric phenomena, the b values significantly less than 1, that indicates a significant coherence of structure-generating processes integrating lower-level hierarchical units to a higher-level entity. For geomorphological structures, both terrestrial and oceanic, the scaling parameters has appeared close to 1 (a "direct transfer" type of generating processes). For plankton systems, which are related with water masses, the b values vary from 1 (processes of direct transfer) up to 2 (random dispersal or diffusion processes). The author attributes this difference to the principle distinctions in dynamic properties of the physical environments for terrestrial (and probably, benthic) and plankton organisms. Finally, for the units (structures or processes) of one and the same organization level, scaling exponents are significantly higher and close to 2 (diffusive dynamics) or more (rigid spatial limitation). Thus, the development of many ecological structures looks dynamically like diffusion or gradual growth, but their putting in the higher-level order (integrity) is a qualitative leap forward and demands appropriate cooperative organizational processes. PMID- 11871267 TI - [Myrmecochorous syndrome]. AB - Myrmecochorous plants have a set of morphological, anatomical, biochemical and phenological features connected to ant dispersal. This complex of plant adaptations is called myrmecochorous syndrome. Present review is based on literature data and original field and laboratory experiments of the authors. The diversity of plant adaptations and its role for ant attraction are analysed. The main feature of myrmecochorous syndrome is elaiosome (a fat body) connected with diaspore. Elaisomes differ by shape, colours, size, anatomy and origin. Different parts of ovule, pericarp or even flower may serve as an initial tissue for elaiosome origin. Fats of elaiosome have a particular complex of fat acids, mainly 1,2-diolein, that attract ant workers. Seed setting periods, synchronizing with maximum ant activity, strictly determined size of diaspores also help in ant attraction. If a plant is not obligate myrmecochore and has some additional mechanism for diaspore dispersal, some characters of myrmecochorous syndrome may be absent or less expressed. PMID- 11871268 TI - [Structure and function of pelagic zooplankton in various types of lakes (using the example of small lakes in northwestern Russia]. AB - Changes of structure and functioning of pelagic zooplankton under lake eutrophication were studied in a group of small (without outlet) lakes in southern Karelia and Leningrad region. The lakes were morphologically similar and located within the same climatic zone. Differences in their trophic status were connected with anthropogenic eutrophication. Correlation between species number and feeding resources of a lake is determined as: Y = (8.01 +/- 3.85) + (0.29 +/- 0.07)X1 + (6.75 +/- 1.52)X2, r2 = 0.95, (1) Y--number of zooplankton species, X1- average chlorophyll concentration for the season, mg/1; X2--average value of biochemical oxygen demand (expressed in mgC/1). Average biomass of zooplankton for season also depends on food: logY = (-0.054 +/- 0.224) + (0.242 +/- 0.094)logX1 + (0.170 +/- 0.179)logX2, n = 13, r2 = 0.87, (2) Y--average zooplankton biomass for the season, kcal/m3, X1 and X2 the same as (1). Increase in species number and zooplankton bimass determined primarily by Rotatoria occur in parallel to chlorophyll concentration and activity of bacteria. In acid lakes rotifers are not important in energy balance. In mezotrophic lakes two main energy paths are formed--through rotifers and through Crustacea. The role of rotifers is extremely important in eutrophic lakes where about 80% of energy paths through them. The ratio community production to energy consumption for the growing season is also depended on the community structure (species number, equitability, connectence): Y = (2.257 +/- 0.026) - (0.368 +/- 0.031)X1 + (5.160 +/- 0.442)X2, r2 = 0.99, (3) Y--average seasonal production of zooplankton, kcal/m2; X1--maximal meaning of Shannon index (bites), calculated on biomass value; X2--connectence of the community, calculated according Briand (1983). Eutrophication changes the stability of lakes relative to external influences, while low productive lakes are very sensitive to the increase in nutrient load, high productive lakes are more influenced by changes in fish predation. PMID- 11871269 TI - Ethical considerations in research: a framework for practice. PMID- 11871270 TI - Prostate-specific-antigen (PSA) testing. PMID- 11871271 TI - A role for physician assistants in organ procurement. AB - Healthcare in the United States, as well as the environment in which physician assistants work, is rapidly changing. Consequently, the role of a physician assistant is expanding to meet the needs of patients and physicians of all specialties of medicine and surgery. In organ procurement, physician assistants can be a valuable asset to an organ procurement organization because of their medical education, versatility, and commitment to personalized care to organ donors and their families. Physician assistants are healthcare professionals who are uniquely qualified for a variety of roles within the organization: clinician, educator, consultant, donor and family liaison, and researcher. PMID- 11871272 TI - Cultural analysis of an organ procurement organization. AB - Organ procurement organizations are a critical link in the acquisition, placement, and transport of human organs for transplantation. Employed within the organ procurement organizations are organ transplant coordinators and support staff who constitute the front line in the challenge to diminish the gap between the supply and demand for organs. Responsibilities are emotionally and physiologically demanding as employees regularly face death and grieving families with empathy and commitment. This study describes and interprets dominant themes embedded in the culture of one such organ procurement organization, geographically located in northeastern United States. The authors used ethnographic research methods to understand the shared meaning system of the members of this organization. Identified themes included coping and sharing, conflicting priorities and uncertainty, and mission and reward. The influence of each theme on the organization is discussed with prescriptive implications for managerial practice. PMID- 11871273 TI - The National Donor Family Council and its giving, grieving, growing program. AB - This article describes the National Donor Family Council of the National Kidney Foundation and its activities on behalf of organ and tissue donor families. Special emphasis is given to the National Donor Family Council's Giving, Grieving, Growing program. PMID- 11871274 TI - Hepatitis B immune globulin: product information and nursing implications. PMID- 11871275 TI - Challenges in donor care. AB - This article is provided as a self-study feature on care of organ donors. Questions and possible responses are provided. Review all the responses for each question. PMID- 11871277 TI - Substance abuse in heart transplant recipients: a 10-year follow-up study. AB - CONTEXT: Active substance abuse is widely accepted as a contraindication for heart transplantation because of the theory that relapse would occur after transplantation and result in compromise of graft function, lowering long-term survival. Listing "recovering" substance abusers for heart transplantation has been controversial. OBJECTIVE: To determine if substance abuse plays an unfavorable role in the outcome of heart transplantation. METHODS: The medical records of all patients at our center who received a heart transplant more than 10 years ago were retrospectively reviewed for history of substance abuse. RESULTS: From a pool of 189 transplant recipients, 47 patients had a definite history of substance abuse (group 1) and were considered in recovery, whereas 142 patients were clearly without any history of substance abuse (group 2). Group 1 had a significantly greater percentage of patients with a pretransplantation diagnosis of idiopathic cardiomyopathy (P = .003), a higher occurrence of heart related cause of death (P = .017), and a significant prevalence of noncompliance (P = .0001) and death because of noncompliance (P = .0004). In contrast, group 2 surprisingly had a significantly higher incidence of death related to infection (P = .0062), which is unexplained. Despite higher incidence of noncompliance in group 1, there was no significant difference in the overall survival rate between the 2 groups. CONCLUSION: These results suggest that patients with a history of substance abuse can undergo successful heart transplantation with acceptable long term survival, though they are at greater risk for substance abuse relapse and resulting noncompliance with the treatment regimen. The extent to which relapse of substance abuse and its consequences affect the cost of posttransplantation care remains to be determined. PMID- 11871278 TI - The economics of kidney transplantation versus hemodialysis. AB - Using 1998 data for 8 transplantation centers located in New York City, this study found that kidney transplantation was a more cost-effective treatment than hemodialysis for the Medicare program. The initially higher costs of transplantation were fully recouped by Medicare 2 years and 10 months after surgery. For persons who are eligible for Medicare solely due to their end-stage renal disease status, transplantation would generate average monthly savings of $3800 over dialysis for the 2 years following the break-even point. For those eligible for reasons other than end-stage renal disease, for example, the aged, the average savings would be $2400. The savings difference arises because Medicare coverage for immunosuppressants for the former group ends 36 months after transplantation. PMID- 11871279 TI - Resources for patients. Infection after transplantation: bugs and drugs. PMID- 11871276 TI - Reducing adverse effects of immunosuppressive agents in kidney transplant recipients. AB - Success in solid organ transplantation with minimal complications can now be achieved for most patients, and a remarkable rate of graft and patient survival can also be expected. However, the potential for adverse events and comorbid conditions increases with longer graft survival. Although the immunosuppressive regimen is central to the outcome of the transplant recipient and directly impacts the survival of the graft, chronic use of immunosuppressive agents is associated with metabolic disturbances such as hypertension, hyperlipidemia, loss of bone density, nephrotoxicity, and diabetes, which may contribute to other comorbid conditions. In addition, changes in appearance, gingival hyperplasia, hirsutism, alopecia, and weight gain disrupt quality of life and may lead to noncompliance with the immunosuppressive regimen. New immunosuppressive medications, including mycophenolate mofetil, sirolimus, basiliximab, and daclizumab, have allowed for experimentation with new regimens designed to reduce or allow discontinuation of corticosteroids and calcineurin inhibitors. This review highlights the impact and cost of immunosuppressive side effects and the potential for new immunosuppressive regimens to reduce this substantial clinical burden in transplantation. PMID- 11871280 TI - Informatics. A new dimension in nursing. PMID- 11871281 TI - Tips on evaluating health care Web sites. PMID- 11871282 TI - How to get great search results on the Internet. PMID- 11871283 TI - Telenursing the career for the 21st century. PMID- 11871284 TI - Nursing technology of the future. PMID- 11871285 TI - My summer in Africa. PMID- 11871286 TI - [Stress-inducible bacterial proteins and virulence]. AB - Different species of pathogenic bacteria, including Salmonella, Neisseria, Listeria and Francisella have been used to demonstrate relationship between the synthesis of stressor induced proteins by cells and the phenotypic manifestation of their virulence. The impact of such external factors as high temperature, low pH, osmolarity, substrate limitation, the content of active forms of oxygen, etc. is accompanied by the synthesis of different stressor induced proteins playing a complex role. Under unfavorable environmental conditions the synthesis of these proteins ensures the survival of the infective agents. Under conditions of a macroorganism synthesis of some stressor induced proteins promotes the survival of infective agents and their resistance to the action of humoral and cell mediated protective factors of the host. As is known, the expression of virulence genes is not constitutive. The expression of these genes greatly depends on environmental conditions and its induction is determined by extra- or intracellular location of the infective agent. Several systems of the regulation of bacterial pathogenicity factors have been described that are relatively not numerous, conservative and respond to external signals. The relevance of a number of stressor induced proteins of bacteria to virulence associated factors is discussed. PMID- 11871287 TI - [Characteristics of immune response in acute intestinal enterobacterial infections]. AB - Update literature on the problem of microorganism--host interactions with pathogenic enterobacteria as a model is reviewed. The activation of lymphoid cells and their protective role is shown to be due to the stimulating action of bacterial antigens. The role of cytokines and stimulation of their expression are discussed. Some phenomena of immunosuppression, phases of enteric mucosal immune response are described. PMID- 11871289 TI - [Outbreaks of acute enteric infections in Moscow in recent years: time of epidemic risk]. PMID- 11871288 TI - [Outbreaks of enteric infections in Moscow in recent years: contingents (institutions) of epidemic risk]. PMID- 11871290 TI - [Salmonella infection in surgical departments of a city hospital]. PMID- 11871291 TI - [The microflora of ulzerous zone mucosa in patients with duodenal ulcer]. AB - Bacteriological study of the biopsies taken from gastric and duodenal mucosa of 10 healthy volunteers and 74 patients with duodenal ulcer, was carried out. In the gastroduodenal zone of healthy subjects microorganisms of 6 genera (Streptococcus, Candida, Staphylococcus, Bacillus, Helicobacter and Lactobacillus) were detected. H. pylori was isolated in 20% of cases only in biopsy specimens taken from the antral section of the stomach of healthy as monoculture or in combination with C. albicans. In patients with duodenal ulcer activation of opportunistic microflora was observed in the periulcerous zone. More often H. pylori occurred in associations with fungi of the genus Candida, streptococci, staphylococci, enterobacteria, Pseudomonas and other microorganisms (of more than 30 genera). Quantitatively the dominating microorganisms (3.8-5.7 lg CFU/g) were H. pylori, fungi of the genus Candida, bacteria of the genera Streptococcus, Peptostreptococcus, Bacteroides, Gemella, Prevotella, Veillonella, Peptococcus, Bacillus, different species of opportunistic enterobacteria, as well as bacteria of the genera Staphylococcus, Micrococcus, Corynebacterium, Neisseria, Pseudomonas, etc. Opportunistic bacteria detected in the ulcerous zone, as a rule, expressed hemolytic, lecithinase, RNAase, caseinolytic, catalase and urease activity. Sonicated filtrates of such cultures produced a cytotoxic effect on cells HEp-2. Ulcer is an infected wound that needs sanitation. PMID- 11871292 TI - [Viral-bacterial-fungal associations in chronic tonsillitis in children]. AB - The microflora of palatal tonsils was studied in 84 children with chronic tonsillitis in comparison with that in the control group of 38 healthy children. In most of the sick children viral-bacterial and less frequently viral-bacterial fungal associations were detected with the prevalence of reo- and adenoviruses, Epstein-Barr viruses, coagulase negative staphylococci and Staphylococcus aureus, as well as peptostreptococci. Adhesive activity and persistence factors among the main bacterial pathogens were shown to be widely prevalent. The depth of the lesion of tonsillar tissue by the infective agents of bacterial and fungal nature, as well as their persistence potential, depended on the taxonomic position of these microorganisms. PMID- 11871293 TI - [Electron microscopy of Mycobacterium leprae passed in laboratory animals]. AB - The ultrastructure of M. leprae was studied in the process of its intraplantar passage from man to mice (passages 1-8). Changes in the morphology of M. leprae, observed in the course of 3 passages, were established. From the phase of dormant forms (passage 1) the bacteria became adapted to the new host by passage 3, which was confirmed by the absence of differences in the ultrastructure of cells by passages 3-8. The possibilities of the phenotypic variability of M. leprae in the process of in vivo passage should be taken into consideration when they are used for the production of specific biopreparations, experimental modeling and the screening of antileprous preparations. PMID- 11871294 TI - [Photoinactivation of microorganisms in the presence of photosensitizers]. AB - Two photosensitizing preparation for the photodynamic therapy of oncological diseases have been obtained. As revealed in this study, under experimental conditions their action suppresses the growth of Enterococcus faecalis and Staphylococcus aureus, which may be of great importance for the treatment of infections. And in contrast to antibiotic therapy, their use may not affect the development of normal intestinal microflora. The study has shown that their derivatives with the residues of carbonic (or muramic) acids more intensively penetrate microorganisms through their cell walls. This makes in possible to expand the spectrum of action of the preparations. The proposed method permits the rapid analysis of the photochemical activity of preparation for photodynamic therapy in the process of their development. PMID- 11871295 TI - [Evaluation of reactogenicity and immunogenicity of Bubo-M, the Russian combined vaccine for immunization of adults against diphtheria, tetanus and viral hepatitis B]. AB - Bubo-M, the first Russian associated vaccine, was found to have low reactogenicity and high immunogenic potency. The frequency of postvaccinal reactions in the group of persons immunized with Bubo-M (20%) appeared to be considerably lower than among persons who received the combined injection of adsorbed DT toxoid with reduced antigen content and vaccine against hepatitis B (47.7%). Following the course of vaccination the level of anti-HBs considerably exceeded the protective level. Immune response to the diphtheria and tetanus components of Bubo-M exceeded that observed after immunization with absorbed DT toxoid with reduced antigen content (p < 0.05). PMID- 11871296 TI - [The state of infectious morbidity in the Russian Federation in 2000 and methods of its stabilization]. PMID- 11871297 TI - [Experimental reactivity and antigenic activity of the bacterial-fungal antigen complexes]. AB - The immunogenic and reactogenic properties of monovaccines prepared from Staphylococcus aureus, S. epidermidis and Candida albicans, as well as those of a associated polyvaccine prepared from these infective agents, were experimentally studied on rabbits. The monovaccines and the associated bacterial-fungal vaccines were found to be safe and faintly reactogenic; in the blood serum of vaccinated rabbits a growth in the titer of agglutinins and its preservation at a high level for 4 months were noted. PMID- 11871298 TI - [Comparative study of the effectiveness of HBcAg presentation to the immune system using attenuated Salmonella strains, serovars S.enteritidis and S.typhimurium]. AB - In this work the results of obtaining HBcAg-producing attenuated Salmonella strains, serovars S. enteritidis and S. typhimurium, and their comparative study is presented. As revealed in this study, attenuated S. enteritidis strain E-23 and S. typhimurium strain T-10, producing HBcAg, induce cell-mediated and humoral immune response to HBcAg after injected into anovals. After injection S. typhimurium strain T-10 induces a much higher titer of specific antibodies than S. enteritidis strain E-23. The level of specific antibodies induced by recombinant HBcAg seems to correlate with the capacity of salmonellae for survival inside macroorganisms. PMID- 11871299 TI - [Vaccination of children under immunosuppressive therapy with ADS-m anatoxin ]. AB - The effectiveness and safety of the immunization of children with adsorbed DT toxoid with reduced component with disturbances in the reactivity of the body, caused by immunosuppressive therapy, are evaluated. The group under study included 53 children with glomerulonephritis, malignant tumors and acute lymphoblast leucosis, formerly receiving immunosuppressive drugs. The control group consisted of 24 patients with pyelonephritis who had not received such therapy. In the postvaccinal period no exacerbation of the main disease was registered in children. The manifestation and frequence of postvaccinal reactions did not differ from those observed in healthy children after the injection of adsorbed DT toxoid with reduced antigen content. Control serological testing demonstrated the presence of pronounced immune response in the vaccinees. PMID- 11871300 TI - [Correction of immune response using purified staphylococcal toxoid and likopid in the secondary immunodeficiency induced by Coxsackie virus B3]. AB - The action of immunomodulators, purified staphylococcal toxoid (PST) and lycopid, on secondary immunodeficiency state developing during infection caused by Coxsackie virus B3 was studied. This defect was manifested by delayed hypersensitivity to sheep red blood cells (SRBC) and the suppression of neutralizing antibodies to poliomyelitis virus. Depending on the scheme of the experiment, PST normalized the defects of immune response to SRBC or poliovirus, increased suppression or showed no activity. Lycopid corrected the defects of humoral response to SRBC. The combination of PST and lycopid was found to produce no increase of suppression. The suggestion was made on the expediency of combination of two (and probably more) immunomodulators for increasing the efficiency of correction of secondary immunodeficiency. PMID- 11871301 TI - [Use of prodigiozan and tactivin for treatment of patients with chronic Klebsiella infections]. AB - Two-stage immunocorrection in patients with chronic Klebsiella infections by consecutive administration of prodigiozan and tactivin, has been shown to facilitate the restoration of the balance of immunoregulating cells, to enhance proliferative response to T- and B-mitogens, thus producing favorable clinical dynamics. The possibility of chronic Klebsiella infections therapy, based on the use of immunocorrecting preparations (prodigiozan and tactivin) and antibiotics capable of intracellular penetration (sumamed and cyprofloxacin) is discussed. PMID- 11871302 TI - [Level of serum antibodies to opportunistic microflora as a marker of the secondary immunodeficiency processes in children]. AB - Examination of children with different noninfectious diseases resulted in obtaining the data base on the state of health of 201 children belonging to the potential risk group of the development of secondary immunodeficiency. The children were subdivided into several groups which differed by the type of immune disturbances and accompanying metabolic shifts. The level of antibodies to one of the fragments of peptidoglycan-N-acetylmuramyldipeptide was compared with the character of changes in the immune system. Different titers of serum antibodies to peptidoglycan were found to correspond to different forms of immune disturbances. The study showed that from the group with the absence of definite signs of immunodeficiency to the group with the pronounced deficiency of T lymphocytes and monocytes expressing CD14 antigen changes in the immune system increased in parallel to a rise in the concentration of alpha 2-macroglobulin, the development of hypercholesterolemia and a decreased level of antibodies to peptidoglycan. Opportunistic microflora was seemingly an important factor in the formation of definite forms of disturbances of the immune system and accompanying metabolic shifts. PMID- 11871303 TI - [Efficiency of immunocorrection using cytokines in the therapy of papillomavirus infection]. AB - The data of the immunological examination and the results of treatment of 86 patients with papillomavirus infection (PVI) are presented. The multifactor suppression of cell-mediated and humoral immunity was established. The degree of immune disturbances correlated with the spread and severity of lesions. The use of systemic injections of cytokines (leukinferon and concentrated interferon) in low doses as adjuvant therapy with laser-radiosurgery led to the normalization of most immunological characteristics and the course of PVI without relapses. PMID- 11871304 TI - [Allergic alteration of leukocytes in patients with drug intolerance]. AB - Increased sensitivity to benzylpenicillin, ampicillin, claforan, cefzol, novocain and lidocaine was determined by means of the leukocyte allergic alteration (LAA) test in 599 patients with intolerance reactions to these preparations in their medical history. In most of these patients the test results exceeded 30% and reached 98%, their average values exceeding those in the control group no subjects with no drug intolerance. The LAA test ensured a high level (83%) of the detection of drug intolerance. PMID- 11871306 TI - [Dysbacteriosis in patients with colon polyposis]. AB - The study revealed the most profound changes in the composition of intestinal microflora in patients with polyposis of the large intestine. In these patients anaerobic microflora (bifidobacteria, lactic acid bacteria) was more often suppressed than in other examined groups, in particular, patients with cholelithic disease. The associations of hemolytic Escherichia with Klebsiella pneumoniae, Pseudomonas aeruginosa, Citrobacter freundii were often observed as well as the increased content of enterococci and fungi of the genus Candida. The determination of frequency and degree of manifestations showed that dysbacteriosis was registered in the absolute majority of patients (97.4%) with polyposis of the large intestine. Among patients with cholelithic disease disturbances in microbiocenosis were detected in 60.0% of cases, the profundity and character of the microflora composition changes being less pronounced. PMID- 11871305 TI - [Identification of hypervariable and conservative fragments of Listeria genome]. AB - The computer analysis revealed hypervariable and highly conservative fractions in the genes of Gram-positive bacteria of the Listeria genus. As a result of analysis of gene iap coding protein p60, PCR based test systems for detection of 6 Listeria species, L. monocytogenes, L. seeligeri, L. ivanovii, L. innocua, L. grayi and L. welshimeri have been developed. Species-specific and conservative gene fragments coding Listeria pathogenicity factors, listeriolysin and cytolysin, were detected. The sets of primers for detection and gene typing of L. monocytogenes, L. seeligeri and L. ivanovii containing cytolysin have been made. The gene typing of Listeria may be carried out in one reaction with the use of multiplex PCR: amplified fragments for different Listeria species differ in the length of the amplified product. The developed sets of primers have a 95-100% degree of homology and may be recommended for the detection and gene typing of Listeria. PMID- 11871307 TI - [Pathogenetic role of intestinal microbiocenosis in the pathogenesis of hereditary myodystrophy]. AB - The effect of the inclusion of probiotic preparations for the correction of disturbances in normal intestinal microflora into the complex therapy of patients wish Duchenne's childhood muscular dystrophy and Becker's myopathy was analyzed. Probiotic therapy made it possible to improve the clinical state of patients, manifested by an increase in muscular strength and accompanied by positive shifts in electromyographic, immunological, biochemical, hormonal characteristics. Intestinal microbiocenosis plays seemingly a certain role in the formation of hereditary pathology. PMID- 11871308 TI - [Effect of medicinal plant extracts on the growth of microorganisms]. AB - Extracts obtained from sweatweed and licorice roots, flax seeds, milfoil, bur marigold, plantain, coltsfoot, nettle, Indian corn stigmas, laminaria produced a stimulating effect on the growth of Candida albicans test strain and Streptococcus pyogenes test strain Dick 1. Sweatweed, licorice, Aerva lanata and violet extracts influenced the growth of Corynebacterium xerosis 1911, while sweatweed, violet, horse-tail, bur-marigold, camomile, plantain, and nettle extracts influenced the growth of shigellae. The stimulating effect could be supposedly produced by biologically active substances contained in medicinal plants (organic acids, alkaloids, carotinoids, vitamins, microelements). Further studies aimed at the identification of substances producing the stimulating effect are planned. PMID- 11871309 TI - [Use of pneumococcal vaccine in patients with renal pathology]. AB - The use of pneumococcal polysaccharide vaccine in patients with chronic renal pathology has proved to be expedient as it prevents the development of pneumococcal infection. However, active immunosuppressive therapy and massive proteinuria result in decreased duration of antibody persistence, which makes in necessary to repeat the injection of the vaccine 20-22 months after the first vaccination. PMID- 11871310 TI - [Effect of lysogenic conversion of Corynebacterium diphtheriae strain on its iron assimilation and spectrum of cellular proteins]. AB - Growth of the isogenic pair of C. diphtheriae strains, the nontoxigenic strain C7 (-) and the toxigenic strain C7 (beta), was studied under conditions of limited availability of the iron source. The growth of the toxigenic strain was shown to depend on the concentration of iron in the medium to a lesser degree than the nontoxigenic one. Lysogenic conversion results in the synthesis of additional iron-dependent proteins, absent in C. diphtheriae initial nontoxigenic strain C7 (-). Special attention was paid to proteins with a mol. wt. 66 kD, synthesized by the toxigenic strain irrespective of the concentration of iron in the medium, while in the toxigenic strain these proteins were detected only under conditions of iron deficiency. PMID- 11871311 TI - [Prognostic model of drug abuse and HIV infection morbidity among youth]. AB - The description of prognostication model the distribution process of a narcotism (the heroine) among youth of large city is given. The model is developed with the purpose of study of a consequence of a narcotism among the teenagers and youth, which use a intravenous drugs user and have high risk of infection by a HIV infection. The results of settlement researches heroine epidemic process in large city with the population about 1 million on retrospective per 10 years are submitted. The forecast of number of drug users on prospect per 5 years is made. It allows to estimate long-term consequences of a narcotism, as "has sunk down" for powerful epidemic AIDS in city in nearest 10 years. PMID- 11871312 TI - [Isolation of a novel rickettsia from the tick Dermacentor silvarum in Baikal lake region]. AB - A new species of rickettsiae with unknown pathogenicity has been detected in ticks Dermacentor silvarum in the region of Baikal Lake. As revealed by the analysis of the primary structure of the gene fragment coding surface membrane protein of 190 kD (rOmpA), the nucleotide sequence of the rickettsiae under study is mostly similar to the sequences of R. sp. MOAa isolate (96%), R. sp. WB-8-2 (96%), R. massiliae strain GS (94%), Rickettsia BAR-29 (94%), R. rhipicephali (94%). Similarity with the sequence of R. sibirica has proved to be 91%. The data thus obtained indicate that the detected rickettsiae represent a new rickettsial species in the territory of East Siberia. PMID- 11871313 TI - [Eltor cholera in Latin America]. AB - Cholera epidemics in South and Central America in the period 1991-1999 are characterized. The mechanisms of existence of Vibrio eltor in environmental objects have been substantiated. The causes and conditions of endemic foci formation which determine the infection spread not only within the countries of the subcontinent, but also far beyond its borders are shown. The effectiveness of cholera control interventions is evaluated. PMID- 11871315 TI - [Clarithromycin in eradication therapy of helicobacter infections in children]. PMID- 11871316 TI - [Diagnosis and treatment of infections caused by Helicobacter pylori]. PMID- 11871317 TI - [Efflux pumps of gram-positive bacteria and resistance to fluoroquionolones]. PMID- 11871314 TI - [Modern methods of detection of HIV resistance to the action of anti-retroviral agents]. AB - The work deals with the molecular mechanisms of the development of resistance of HIV to modern preparations for treatment of HIV infection and AIDS: inhibitors of reverse transcriptase and protease. The principles and concrete schemes of the use of modern methods of geno- and phenotypic resistance determination of HIV are described in detail. The comparative characterization of both approaches together with the problems and difficulties connected with the interpretation of the results of resistance tests are presented, their probable clinical importance is discussed. Practical recommendations concerning the use of tests for the determination of HIV resistance under different clinical conditions, based on the conferences and literature data on HIV infection and for the period of 1999-2000, are reviewed. PMID- 11871318 TI - [Basic results of studying antimicrobial agents of the quinolone class at the beginning of the XXI century: successes and failures in developing highly effective fluoroquinolones]. PMID- 11871319 TI - [Evaluation of the effectiveness of antibacterial substances in treating an experimental form of bubonic plague in monkeys]. AB - The modelling of glandular plague and selection of the conditions for estimating the efficacy of new antibacterials for the treatment of the infection were performed on hamadryads (baboons). The experiments showed that the average LD50 of the culture of a highly virulent strain of Yersinia pestis on its subcutaneous administration to the animals was 2089 viable microbes. In 18 per cent of the episodes the experimental glandular plague in the animals was complicated by secondary plague pneumonia. Subcutaneous administration of 2 x 10(7) viable microbial cell of the plague pathogen caused acute sepsis and the animal death. The treatment of the experimental glandular plague in the hamadryads demonstrated that new antibacterials such as amikacin, netilmicin, ceftriaxone, cefotaxime, ceftizoxime, doxycycline, rifampicin, ofloxacin and ciprofloxacin were not inferior in their efficacy to streptomycin and tetracycline successfully used in the therapy of patients with plague. PMID- 11871320 TI - [Combination chemotherapy of drug-resistant tuberculosis with inclusion of ofloxacin (zanocin)]. PMID- 11871321 TI - [Forensic-medical evaluation of the histologic structure of the hymen]. AB - Variants of microscopic structure of the hymen are described and its histological classification based on the stromal component structure is offered. Hypotheses on the hymen histogenesis under different conditions are formulated. Histological analysis and histological classification of the hymen are needed for objective solution of questions arising during expert evaluation of virginity and its violation, probability or improbability of coitus without defloration. PMID- 11871322 TI - [Forensic-medical diagnosis of heart damage from blunt trauma]. AB - A total of 277 forensic expert medical conclusions on cases with closed heart injury are analyzed. The morphology and mechanisms of formation of heart injuries in various traumas are discussed. PMID- 11871323 TI - [Some aspects of forensic-medical studies of dog bite injuries]. AB - The incidence of dog attacks of humans has increased, but no publications or methodological recommendations on this problem appeared. The history of the problem and statistical data reflecting distribution of injuries with regard to sex and age, location of injuries, and time of the day are presented. Signs which help differentiate injuries inflicted by dog and human teeth or injuries with blunt objects are defined. Further investigation of the problem is needed. PMID- 11871324 TI - [Detection and assessment of microparticles in the area of periosteum damage from blunt trauma]. AB - Periosteal injuries and deposition of trace corpuscles on the periosteum resultant from stroke with blunt hard objects and in car or railway accidents are analyzed. The greatest number of trace corpuscles of different origin are formed as a result of direct strike of a hard blunt object against the bone surface. They are deposited at the site of periosteal injury, which should be removed and examined by special methods. This will help determine the mechanism of injury and identify the object with which the injury was inflicted. PMID- 11871326 TI - [Study of an electrical charge self-defense device]. AB - An electric charge device for self defense was evaluated on a volunteer. The device is safe for life and health of a healthy adult human. Moral aspects of such trials on humans are discussed. PMID- 11871325 TI - [Study of human salivary agglutinins in expert evaluation of disputable paternity]. AB - Analysis of the saliva for evaluation of AB0 production should be carried out along with other studies in expert evaluation of disputable paternity and maternity and in cases with replacement of children. A total of 249 subjects (79 families) were examined. Salivary specimens from 72 families were analyzed; salivary specimens were not analyzed in 7 families in which both men and women had blood group IV (AB0) and no agglutinins were present in the serum. The production of isoantibodies alpha and beta is inherited similarly as the production of AB0 isoantigens. The majority (81.6%) of children whose parents were isoantibody producers, were isoantibody producers too. If one parent produced isoantibodies and the other not, the children were more often (68.1%) producers, but less often than in the previous category. If both parents were non producers, the children were non-producers as well. PMID- 11871328 TI - [M. I. Avdeev--a prominent figure in domestic forensic medicine (on the 100th anniversary of his birth)]. PMID- 11871329 TI - [Distribution of 4-methylphenol in warm blooded animals]. AB - The distribution of 4-methylphenol in warm-blooded animals (rabbits) was studied after intragastric administration of the toxin. 4-methylphenol (free or bound) is present largely in the urine, liver and kidney tissues. PMID- 11871327 TI - [Optimal conditions for extraction of "caffetin" and "saridon" tablet components from aqueous solutions]. AB - Using a mathematical method of experiment planning (Latin square), the authors suggest the optimal conditions for extraction of propifenasone and paracetamol, the basic components of caffeine and saridon tablets, from water solutions: extraction with ethylacetate (pH 2) for 5 min in the presence of an electrolyte (sodium chloride or ammonium sulfate) quantum satis. The possibility of extraction of caffeine and codeine under these conditions was tested. When extracting the components of caffeine and saridon tablets, paracetamol, propifenasone, and caffeine should be extracted with ethylacetate at pH 2 and codeine by chloroform at pH 10. PMID- 11871330 TI - [Gas chromatographic method of quantitative determination of carbon monoxide in cadaver blood]. AB - A gas chromatographic method has been developed for measurement of carbon monoxide in cadaveric blood on a chromatographer with thermal conductivity detector and air oxygen as the internal standard. The error of the method is no more than 4% with the mean quadratic deviation +/- 0.18-0.33 min. The method was studied on 10 model and 30 expert blood samples. Putrefactive products did not influence the accuracy of analysis. PMID- 11871331 TI - [Analysis of forensic-medical expert conclusions on rap and concealed rape (data from anonymous questionnaires)]. AB - Specific features of modern cases of raping are analyzed on the basis of the data of forensic medical obstetrical and gynecological expert evaluations carried out in the Primorye territory. Age and risk groups are defined, health status of victims is characterized, and specific injuries of the body and genitals are described. Specific features of concealed sexual violations in Vladivostok are defined and the need in creation of rehabilitation centers for victims of raping is discussed. PMID- 11871332 TI - [S. F. Gaevskii--forensic medic and organizer of health care in Russia]. PMID- 11871333 TI - [Methods for analyzing synthetic pyrethroids]. PMID- 11871334 TI - [Clinico-morphological parameters of heroin abuse and pathology connected with it]. AB - Morphological manifestations of narcomania in heroin users are described. Diseases associated with heroin narcomania and causes of death of heroin users are presented. Morphological manifestations at the site of injections and inflammatory reactions in parenchymatous organs are characterized. The authors pay special attention to the productive inflammation which can serve as one of the signs that confirm narcomania in cases with parenteral injections of crude heroin. The authors refer the granulomas to toxic allergic form of granulomatous hypersensitive inflammation. PMID- 11871335 TI - Proteomics. Special issue dedicated to Professor J. Klose. PMID- 11871336 TI - Molecular replacement and its relatives. Proceedings of the 2001 CCP4 study weekend. PMID- 11871337 TI - Mediterranean Diet. Proceedings of the 13th International ISFE (International Foundation for the Promotion of Nutrition Research and Nutrition Education) Symposium. September 23-25, 1999. Torino, Italy. PMID- 11871338 TI - Individual monitoring of external radiation. Proceedings of a European Workshop. Helsinki, Finland. September 4-6, 2000. PMID- 11871339 TI - Women and Modern Medicine. Proceedings of a symposium. 10-11 November 1994. PMID- 11871340 TI - Current developments in anaesthesia. Proceedings of a symposium, Barcelona, February 2001, and abstracts from the 7th International Neuromuscular Meeting, Belfast, June 2001. PMID- 11871341 TI - Abstracts of the 83rd Annual Meeting of the American Radium Society. April 26-29, 2001. PMID- 11871342 TI - Results of surgical treatment for faecal incontinence. PMID- 11871343 TI - 3rd Fall Meeting of the Sexual Medicine Society of North America, Inc. December 7 9, 2001. Charleston, South Carolina, USA. Proceedings and abstracts. PMID- 11871344 TI - Ongoing clinical trials. PMID- 11871345 TI - Bibliography. Current world literature. Cataract surgery and lens implantation. PMID- 11871346 TI - The melatonin hypothesis: a matter of method. PMID- 11871347 TI - New mining regs, new discontent. PMID- 11871348 TI - The price of bottled water. PMID- 11871349 TI - A better way to water. PMID- 11871350 TI - United Nations Convention to Combat Desertification. PMID- 11871352 TI - Abstracts of the annual meeting of the Association for Academic Minority Physicians. October 12-14, 2001. PMID- 11871351 TI - Romanian Conference of Immunology and Allergology. Abstracts. PMID- 11871354 TI - Nuclear medicine and PET: Frost & Sullivan see gains. PMID- 11871353 TI - Saliva in health and disease. Proceedings of the 16th International Conference on Oral Biology. April 9-12, 2000. Chantilly, Virginia, USA. Proceedings. PMID- 11871355 TI - Physical activity and mortality among women. PMID- 11871356 TI - Low intensity Nd:YAG laser irradiation for lateral epicondylitis. PMID- 11871357 TI - [Management of estrogen deficiency in women after breast cancer therapy]. PMID- 11871358 TI - [83rd Congress of the Association of Morphologists. 23-26 May 2001, Lyon, France. Abstracts]. PMID- 11871359 TI - A school-based asthma intervention program in the Buffalo, New York, schools. AB - This project investigated the feasibility and effectiveness of a school asthma program in reducing asthma exacerbations among school children. In 1997-1998, two schools were selected for a case control pilot study. The intervention required that students with asthma, who needed medication daily at school, must present a written plan from the health care provider. Students with asthma were identified through parent/guardian or school reports. The pilot program was expanded into five schools in 1998-1999. All schools kept records of rescue treatments for asthma episodes. The pilot intervention resulted in an 80% decrease in rescue treatments from 1996-1997 to 1997-1998. In the additional five schools, an overall decrease of 17% occurred in rescue treatments during 1998-2000. Overall, 65% of physicians provided requested Asthma Care Plans (ACP). In two schools, the number of asthma care plans that required anti-inflammatory medications tripled. Preliminary results indicate this school asthma program proved feasible and effective in reducing the frequency of asthma exacerbations at school. PMID- 11871360 TI - Patient compliance with a rehabilitation program after flexor tendon repair in zone II of the hand. AB - The use of early mobilization during rehabilitation of a finger after flexor tendon repair in zone II has achieved worldwide acceptance. Different exercise instructions are given to patients in different clinics, however. When using Kleinert dynamic splinting, some hand therapists instruct patients to exercise 10 times an hour, while others tell their patients to exercise as many times as possible. How often a patient should exercise to achieve optimal results has never, to the authors' knowledge, been investigated. To identify a relationship between exercise behavior and range of motion achieved at 6 months postoperatively, the authors started a pilot study of the compliance of 15 patients with a rehabilitation program following flexor tendon repair in zone II. A portable rehabilitation device was used to deliver an isotonic tensile force and to record when (during the day) a patient exercises and how many exercises the patient performs. Patients were instructed to perform at least ten exercises per hour. The average count per hour was 69 exercises (range, 14-213). In this small group study, no strong correlation was found between exercise count and the total active or total passive motion achieved. PMID- 11871361 TI - Three-dimensional reconstruction of microstructures in gibbsite using charge contrast images. AB - An iterative process of serial sectioning and charge contrast imaging has been applied to gibbsite particles embedded in epoxy resin. The features observed in the two-dimensional sections have been reconstructed into three-dimensional (3-D) objects, so that the volumetric relationships in the microstructure are visualised. The 3-D objects confirm that the image detail present in charge contrast images of gibbsite is related to the microstructure and the processes occurring during the crystallisation of gibbsite. Speculative explanations for what these microstructures may represent are presented. PMID- 11871362 TI - Determination of the reliability and validity of the Marijuana Craving Questionnaire using imagery scripts. AB - The purpose of this study was to determine the reliability and validity of the Marijuana Craving Questionnaire (MCQ) by using active imagery of auditorily presented scripts. Current marijuana users (n = 48) imagined scripts that varied in amount of descriptors of desire to smoke marijuana, from no-urge to high-urge content. Self-reported marijuana craving significantly increased as a function of script-urge intensity on Factors 1, 3, and 4 of the MCQ. Homogeneity of items comprising each MCQ factor was examined, indicating no significant departures from unidimensionality. These results verify and extend the reliability and validity of the MCQ as a multidimensional measurement of marijuana craving. The data also suggest that drug craving is not an all-or-none phenomenon. PMID- 11871363 TI - Improved health outcomes in patients with COPD during 1 yr's treatment with tiotropium. AB - Tiotropium, a novel once-daily inhaled anticholinergic, has been shown to improve lung function over a 24-h period. In order to extend these findings, health outcomes were evaluated over 1 yr in chronic obstructive pulmonary disease (COPD) patients. Spirometric results, peak expiratory flow rate (PEFR), salbutamol use and effects on dyspnoea, health-related quality of life and COPD exacerbations were assessed in two identical 1-yr randomized double-blind double-dummy studies of tiotropium 18 microg once daily (n=356) compared with ipratropium 40 microg q.i.d. (n=179). Screening forced expiratory volume in one second (FEV1) were 1.25+/-0.43 L (41.9+/-12.7% of the predicted value) (tiotropium) and 1.18+/-0.37 L (39.4+/-10.7% pred) (ipratropium). Trough FEV1 at 1 yr improved by 0.12+/-0.01 L with tiotropium and declined by 0.03+/-0.02 L with ipratropium (p<0.001). Significant improvement in PEFR, salbutamol use, Transition Dyspnea Index focal score, and the St George's Respiratory Questionnaire total and impact scores were seen with tiotropium (p<0.01). Tiotropium reduced the number of exacerbations (by 24%, p<0.01), and increased time to first exacerbation (p<0.01) and time to first hospitalization for a COPD exacerbation (p<0.05) compared with ipratropium. Apart from an increased incidence of dry mouth in the tiotropium group, adverse events were similar between treatments. Tiotropium was effective in improving dyspnoea, exacerbations, health-related quality of life and lung function in patients with chronic obstructive pulmonary disease, and exceeds the benefits seen with ipratropium. The data support the use of tiotropium once-daily as first-line maintenance treatment in patients with chronic obstructive pulmonary disease. PMID- 11871364 TI - Potential masking effect on dyspnoea perception by short- and long-acting beta2 agonists in asthma. AB - Asthma patients evaluate the effect of medication treatment through the degree of their asthma symptoms, which might be affected by their ability to perceive these symptoms. It has been suggested that beta2-agonists may mask the effects of an increase in airway inflammation. This study compared the perception of histamine induced bronchoconstriction during monotherapy with short- or long-acting beta2 agonists. Asthmatic patients (68 male and 60 female, mean age 35+/-11 yrs, forced expiratory volume in one second (FEV1) 86+/-15% of the reference value, provocative concentration causing a 20% fall in FEV1 (PC20) geometric mean 0.97 mg x mL(-1) (95% confidence interval (CI): 0.73-1.30)) were selected and randomly allocated to use either a short-acting (salbutamol, n=41) or long-acting beta2 agonist (formoterol, n=46) or placebo (n=41) for 12 weeks. Perception of dyspnoea provoked by histamine-induced bronchoconstriction was measured at the start and every 4 weeks thereafter. Subjects quantified their sensation of breathlessness during the challenge tests on a modified Borg scale at the start of the study and every 4 weeks thereafter. The sensitivity to changes in FEV1 was analysed by the linear regression slope (alpha) Borg versus % fall in FEV1. The absolute perceptual magnitude (PS20) was determined by the perception score at the 20% fall in FEV1. Although the geometric mean PC20 decreased significantly within the group using short-acting beta2-agonists (in the group with initial PC20 > or = 2 mg x mL(-1) there was a drop from 5.26-1.94 mg x mL(-1); p=0.013), repeated measurement analysis showed no difference in the course of time of perception (both slope alpha and PS20) between the three medication groups. This study showed that chronic use of short- or long-acting beta2-agonists in asthmatics for a period of 12 weeks, did not significantly change the perception of histamine induced bronchoconstriction compared with placebo. Further investigation is required to establish whether this suggests that these drugs do not mask a deterioration of asthma. PMID- 11871365 TI - Outgrown asthma does not mean no airways inflammation. AB - Although some asthmatic children seem to recover from their asthma, 30-80% develop asthma again in later life. The underlying risk factors are unknown. The hypothesis for this study was that children with apparently outgrown asthma would have underlying airway inflammation. Nonbronchoscopic bronchoalveolar lavage was performed on normal children (n=35) and children who had wheezed previously (n=35). Eosinophils were raised in the lavage fluid of atopic children who had apparently outgrown asthma (median (interquartile range) 0.36 (0.05-0.74) compared to controls 0.10 (0-0.18), p=0.002). There was no relationship between length of remission and degree of airways eosinophilia. Thus, there is persistent airways inflammation in some children with outgrown asthma and this may be a risk factor for future relapse. PMID- 11871366 TI - Individual allergens as risk factors for asthma and bronchial hyperresponsiveness in Chinese children. AB - The role of allergen sensitization in the development of asthma in the Chinese is not clear. This study aims to determine the relationship of sensitization to individual allergens, and the development of asthma and bronchial hyperresponsiveness (BHR) in schoolchildren from three Chinese cities: Hong Kong, Beijing and Guangzhou. Community-based random samples of 10-yr-old schoolchildren from three Chinese cities were recruited for study using the International Study of Asthma and Allergies in Childhood (ISAAC) Phase II protocol. Subjects were studied by parental questionnaires (n=10,902), skin-prick tests (n=3,479), and methacholine challenge tests (n=608). The prevalence rates of wheeze in the past 12 months (Hong Kong, 5.8%; Beijing, 3.8%; Guangzhou, 3.4%) and atopy (Hong Kong, 41.2%; Beijing, 23.9%; Guangzhou, 30.8%) were highest in schoolchildren from Hong Kong. Multivariate logistic regression analyses revealed that sensitization to Dermatophagoides pteronyssinus (odds ratio (OR)=4.48; 95% confidence interval (CI): 3.02-6.66), cat (2.59; 1.67-4.03), Dermatophagoides farinae (2.41; 1.65 3.51) and mixed grass pollen (2.85; 1.24-6.50) were significantly associated with current wheeze. Atopy, defined as having > or = 1 positive skin-prick tests, was not an independent risk factor for current wheeze in children from any of the three cities. Furthermore, atopy (OR=2.53; 95% CI: 1.07-5.97), sensitization to cat (3.01; 1.39-6.52) and D. farinae (3.67; 1.93-6.97) were significantly associated with BHR. The authors confirmed that sensitization to house dust mite and cat was significantly associated with current wheeze and bronchial hyperresponsiveness in Chinese schoolchildren. However, the difference in the prevalence rate of atopic sensitization cannot explain the higher prevalence of childhood asthma in Hong Kong, when compared with those children from Beijing and Guangzhou. PMID- 11871369 TI - Qualitative assessment of esophageal pH recordings. AB - Quantitative analysis of reflux episodes (the pH score) was used to diagnose pathologic reflux. The aim of the present study was to determine the pattern and significance of variables that may affect the pH scores. The quality of the first esophageal pH recordings were analysed and its effects on the pH scores as calculated by computer were determined. The most common abnormality 13/123 (10.6 per cent) was related to disconnection of the probe or the reference electrode from the pH meter resulting in falsely high pH scores. Abnormalities suggesting incorrect location of the probe occurred in 12/123 (9.7 per cent) resulting in falsely high or low pH scores. Finally, pH curve drift was found in 6 (4.9 per cent) of the recordings. It is concluded that the calculated pH score by computer may be misleading. Therefore, the quality of the pH recording should be determined before interpretation of pH measurement. PMID- 11871368 TI - Impact of vitamin A supplementation through different dosages of red palm oil and retinol palmitate on preschool children. AB - Red palm oil (5 ml and 10 ml), ground nut oil fortified with 400 and 800 retinol equivalent retinol palmitate, and ground nut oil (5 and 10 ml), were administered to six groups of preschool children (four experimental and two control groups) in randomly assigned balwadis of Ramanathapuram District of Tamil Nadu for a period of 7 months, to monitor the difference in the efficacy of the mode of supplementation and the optimum dose for improving vitamin A status. Results show that red palm oil groups recorded more gain in retinol and beta-carotene levels compared to other dosage groups, and that administration of 10 ml did not offer any substantial improvement over the 5-ml daily dose. PMID- 11871367 TI - Chemokines, innate and adaptive immunity, and respiratory disease. AB - Selective leukocyte trafficking and recruitment is primarily regulated by a specific family of small proteins called "chemokines". This extended family shepherds and guides leukocytes through their lives, facilitating their development, regulating their interactions with other leukocyte types, and guiding their recruitment to sites of inflammation. Through the actions of chemokines, allergen sensitization is regulated in atopic asthma, through the controlled migration of dendritic cells, T- and B-lymphocytes, mast cells and basophils. Subsequently, atopic inflammation is driven by chemokine-directed recruitment of eosinophils, basophils and lymphocytes. Diseases from cancer to chronic obstructive pulmonary disease to interstitial fibrosis are all potential targets for chemokine receptor antagonism. Innate immunity (the early pattern recognition responses to stimuli such as lipopolysaccharide, viral proteins and bacterial DNA) needs to bridge the gap to specific immunity and antibody production and immunological memory. Again, chemokines are likely to be fundamental mediators of these responses. Chemokines are fundamental regulators of leukocyte homeostasis and inflammation, and their antagonism by small molecule chemokine receptor antagonists may be of enormous importance in the future treatment of human respiratory disease. PMID- 11871370 TI - Blood cultures from Brazilian pediatric outpatients with community-acquired pneumonia (CAP). PMID- 11871372 TI - Sustained attention in young people at high risk for schizophrenia. AB - BACKGROUND: Sustained attention has been found to be impaired in individuals suffering from schizophrenia and their close relatives. This has led to the hypothesis that impaired sustained attention is an indicator of vulnerability to schizophrenia. METHODS: The Edinburgh High Risk Study used the Continuous Performance Test-Identical Pairs version (CPT-IP) to assess sustained attention in 127 high risk participants, 30 controls and 15 first-episode schizophrenic patients. A second assessment was completed by 59 high risk and 18 control participants 18 months to 2 years after the first. RESULTS: No differences in attentional capacity were found between the high risk and control groups and there was no association between genetic liability to schizophrenia and poor performance on the CPT-IP. Additionally, no association between occurrence of psychotic symptoms in the high risk group and impaired attentional capacity was found. CONCLUSIONS: The results suggest that deficits in sustained attention are not indicative of a genetic vulnerability to schizophrenia, and are not associated with the occurrence of psychotic symptoms. PMID- 11871371 TI - Genetic and environmental contributions to cannabis dependence in a national young adult twin sample. AB - BACKGROUND: This paper examines genetic and environmental contributions to risk of cannabis dependence. METHOD: Symptoms of cannabis dependence and measures of social, family and individual risk factors were assessed in a sample of 6265 young adult male and female Australian twins born 1964-1971. RESULTS: Symptoms of cannabis dependence were common: 11.0% of sample (15.1% of men and 7.8% of women) reported two or more symptoms of dependence. Correlates of cannabis dependence included educational attainment, exposure to parental conflict, sexual abuse, major depression, social anxiety and childhood conduct disorder. However, even after control for the effects of these factors, there was evidence of significant genetic effects on risk of cannabis dependence. Standard genetic modelling indicated that 44.7% (95% CI = 15-72.2) of the variance in liability to cannabis dependence could be accounted for by genetic factors, 20.1% (95% CI = 0-43.6) could be attributed to shared environment factors and 35.3% (95% CI = 26.4-45.7) could be attributed to non-shared environmental factors. However, while there was no evidence of significant gender differences in the magnitude of genetic and environmental influences, a model which assumed both genetic and shared environmental influences on risks of cannabis dependence among men and shared environmental but no genetic influences among women provided an equally good fit to the data. CONCLUSIONS: There was consistent evidence that genetic risk factors are important determinants of risk of cannabis dependence among men. However, it remains uncertain whether there are genetic influences on liability to cannabis dependence among women. PMID- 11871375 TI - Measuring and modeling zinc and cadmium binding by humic acid. AB - Free metal ions in aqueous and terrestrial systems strongly influence bioavailability and toxicity. Most analytical techniques determine the total metal concentration, including the metal ions bound by dissolved organic matter. Ion activity can be measured with ion-specific electrodes (ISEs) for some metals, but an electrode for Zn is not commercially available. As a result, very few data are available on Zn binding by natural dissolved organic matter. The aim of this study is to determine free Zn concentrations in purified humic acid solutions using the recently developed Donnan membrane technique. However, several analytical aspects of the Donnan membrane technique had to be clarified before reliable data could be composed. Cd was chosen for validation. This study shows that free Cd concentrations as measured by the Donnan membrane technique agreed well with Cd ISE measurements. It is also shown that the Donnan membrane technique could be used at high pH. The Donnan membrane technique provided consistent results in a range of p[Cd2+] = 3-9 and p[Zn2+] = 3-8 at pH 4, 6, and 8. Metal speciation in humic acid solutions was also calculated with the consistent NICA-Donnan model using generic parameters. The model could excellently describe the experimental data without adjusting any of the parameters (R2Cd = 0.971, R2Zn = 0.988). PMID- 11871374 TI - CT findings of intracranial blastomycosis in a dog. AB - Computed tomography (CT) findings in a dog with intracranial blastomycosis were marked periventricular contrast enhancement of the lateral ventricles, the 3rd ventricle, and the mesencephalic aqueduct. The CT appearance correlated with the histopathologic findings, where severe ependymitis was present throughout the ventricular system and there was stenosis of the mesencephalic aqueduct due to an inflammatory infiltrate. CT is therefore recommended as a screening test for intracranial blastomycosis in dogs and also as an imaging modality for follow-up evaluation after treatment. This is particularly true in dogs with systemic or ocular blastomycosis, which appear to be at higher risk of developing CNS involvement. PMID- 11871373 TI - Psychosocial work characteristics as risk factors for the onset of fatigue and psychological distress: prospective results from the Maastricht Cohort Study. AB - BACKGROUND: Prolonged fatigue has recently attracted attention in occupational (mental) health research since it may lead to sickness absenteeism and work disability. To date, little is known about the role of psychosocial work characteristics in the aetiology of fatigue. In this study we examined prospectively a wide range of psychosocial work characteristics as possible risk factors for the onset of fatigue and psychological distress in the working population. METHODS: This study is based on 8833 employees, participating in the Maastricht Cohort Study of 'Fatigue at Work'. A wide range of psychosocial work characteristics, measured at baseline, was used to predict the onset of fatigue and psychological distress 1 year later. Fatigue was measured with the Checklist Individual Strength; the General Health Questionnaire was used to measure psychological distress. RESULTS: The cumulative incidence of fatigue during 1 year follow-up was 9.7% (N = 492) in men, and 13.5% (N = 241) in women. Psychological demands at work as well as physical and emotional demands increased the risk for fatigue in men, whereas decision latitude in men and co-worker social support in women were protective against fatigue. These prospective associations remained significant after adjustments for potential confounders and baseline fatigue. As regards psychological distress, no association was found with decision latitude, while conflicts at work increased the risk of psychological distress. CONCLUSIONS: Psychosocial work characteristics were significant predictors for the onset of fatigue in the working population. The prospective associations suggest some differential effects in the aetiology of fatigue and psychological distress. Good interpersonal relationships at work and high decision authority were demonstrated to be relevant aspects that should be targeted for prevention. PMID- 11871376 TI - Aspartame optical biosensor with bienzyme-immobilized eggshell membrane and oxygen-sensitive optode membrane. AB - An aspartame optical biosensor has been fabricated by employing a bienzyme system composed of alpha-chymotrypsin and alcohol oxidase immobilized onto an eggshell membrane and an oxygen-sensitive optode membrane as the transducer. The detection schemes involve the enzymatic reactions of aspartame leading to the depletion of the oxygen level of the medium with a concomitant enhancement of the fluorescence intensity of the oxygen-sensitive membrane. The scanning electron and transmission electron micrographs show the microstructure of the eggshell membrane which is successfully immobilized with bienzyme. Using this novel immobilization technique, the aspartame biosensor shows extremely good stability with a shelf life of at least 8 months. The rate change of the fluorescence intensity in 4 min is found to be linearly related to the concentration of aspartame. The useful analytical working range of the biosensor is from 0.056 to 3.07 mM aspartame. The effects of temperature, pH, and ionic strength on the response of the aspartame biosensor are investigated in detail. Citric acid, cyclamic acid, D-fructose, D-galactose, D-glucose, hydrogen peroxide, DL-malic acid, L-phenylalanine, saccharin, sodium benzoate, and sucrose show no interferences but ethanol interferes strongly. The aspartame biosensor has been applied to determine aspartame contents in some commercial products. PMID- 11871377 TI - Personality and the genetic risk for alcohol dependence. AB - The extent to which the genetic risk for alcohol dependence (AD) and conduct disorder (CD) and their common genetic risk overlap with genetic factors contributing to variation in dimensions of personality was examined in a study of 6,453 individuals from 3,383 adult male and female same-sex and unlike-sex twin pairs from the Australian Twin Registry. The associations between the personality dimensions of positive emotionality, negative emotionality, and AD and CD risk were modest, whereas the associations between behavioral undercontrol and AD and CD risk were substantially higher. Genetic influences contributing to variation in behavioral undercontrol accounted for about 40% of the genetic variation in AD and CD risk and about 90% of the common genetic risk for AD and CD. These results suggest that genetic factors contributing to variation in dimensions of personality, particularly behavioral undercontrol, account for a substantial proportion of the genetic diathesis for AD and most of the common genetic diathesis for AD and CD among both men and women. PMID- 11871378 TI - Radical excision and reconstruction of chronic tibial osteomyelitis with microvascular muscle flaps. AB - We report a series of 21 patients with chronic osteomyelitis of the tibia treated with microvascular muscle flap reconstruction. All patients underwent a radical bone and soft-tissue excision until healthy, well-bleeding tissue was exposed. Six patients required cancellous bone grafting. Latissimus dorsi was used in 14 patients, gracilis in 4, and rectus abdominis in 4. One gracilis flap was lost due to vessel thrombosis and was replaced with a rectus abdominis free flap. Average follow-up was 2.5 years. There was no evidence of clinical infection in 20 patients at follow-up; the bone had healed, the soft-tissue cover was stable, and the laboratory parameters were normal. Bone infection recurred in 1 patient, resulting in a below-knee amputation. The radical excision of infected bone and affected soft tissue and reconstruction with a well-vascularized large free muscle flap is an excellent solution in most difficult chronically infected cases. PMID- 11871379 TI - Platelet concentrate increases bone ingrowth into porous hydroxyapatite. AB - Platelets contain growth factors that are believed to stimulate early fracture repair. Autologous platelets can be sequestered, concentrated, and mixed with thrombin to yield a so-called autologous growth factor gel, which might enhance bone repair or bone graft incorporation. The effect of this platelet concentrate on total tissue and bone ingrowth into porous coralline hydroxyapatite was studied in a bone chamber rat model. Chambers with the platelet concentrate showed a significant increase in bone and total tissue ingrowth distance compared to untreated controls, indicating a platelet concentrate might enhance the clinical performance of porous hydroxyapatite in bone replacement. PMID- 11871380 TI - Multigroup discrete ordinates modeling of 125I 6702 seed dose distributions using a broad energy-group cross section representation. AB - Our purpose in this work is to demonstrate that the efficiency of dose-rate computations in 125I brachytherapy, using multigroup discrete ordinates radiation transport simulations, can be significantly enhanced using broad energy group cross sections without a loss of accuracy. To this end, the DANTSYS multigroup discrete ordinates neutral particle transport code was used to estimate the absorbed dose-rate distributions around an 125I-model 6702 seed in two dimensional (2-D) cylindrical R-Z geometry for four different problems spanning the geometries found in clinical practice. First, simulations with a high resolution 210 energy groups library were used to analyze the photon flux spectral distribution throughout this set of problems. These distributions were used to design an energy group structure consisting of three broad groups along with suitable weighting functions from which the three-group cross sections were derived. The accuracy of 2-D DANTSYS dose-rate calculations was benchmarked against parallel Monte Carlo simulations. Ray effects were remedied by using the DANTSYS internal first collision source algorithm. It is demonstrated that the 125I primary photon spectrum leads to inappropriate weighting functions. An accuracy of +/-5% is achieved in the four problem geometries considered using geometry-independent three-group libraries derived from either material-specific weighting functions or a single material-independent weighting function. Agreement between Monte Carlo and the three-group DANTSYS calculations, within three standard Monte Carlo deviations, is observed everywhere except for a limited region along the Z axis of rotational symmetry, where ray effects are difficult to mitigate. The three-group DANTSYS calculations are 10-13 times faster than ones with a 210-group cross section library for 125I dosimetry problems. Compared to 2-D EGS4 Monte Carlo calculations, the 3-group DANTSYS simulations are a 100-fold more efficient. Provided that these efficiency gains can be sustained in three-dimensional geometries, the results suggest that discrete ordinates simulations may have the potential to serve as an efficient and accurate dose-calculation algorithm for low-energy brachytherapy treatment planning. PMID- 11871381 TI - An investigation of the operating characteristics of two PTW diamond detectors in photon and electron beams. AB - The dosimetric properties of two PTW Riga diamond detectors type 60003 were studied in high-energy photon and electron therapy beam. Properties under study were current-voltage characteristic, polarization effect, time stability of response, dose response, dose-rate dependence, temperature stability, and beam quality dependence of the sensitivity factor. Differences were shown between the two detectors for most of the previous properties. Also, the observed behavior was, to some extent, different from what was reported in the PTW technical specifications. The necessity to characterize each diamond detector individually was addressed. PMID- 11871382 TI - Alternative approaches in IVF. AB - Various new developments in clinical and basic science which may impact on IVF in the near or distant future will be discussed in this review. These key areas include the regulation of early follicle development and the extended in-vitro culture of oocytes and embryos. Moreover, alternative compounds and ovarian stimulation protocols will be discussed, along with highlights in the development of the cryopreservation of excess oocytes or embryos. Finally, the health economics of IVF is addressed. PMID- 11871383 TI - Effects of two preparations of 75-mg extended-release aspirin on platelet aggregation, prostanoids and nitric oxide production in humans. AB - OBJECTIVE: The lowest dose of aspirin shown to be effective in the secondary prevention of thrombotic accidents is 75 mg/day. Presystemic acetylation of cyclooxygenase and the formation of salicylic acid in the liver are fundamental to ensure optimum antithrombotic effects of aspirin. This study was designed to compare the effects of two forms of extended-release aspirin (at 75 mg/day) on prostanoid and nitric oxide synthesis in healthy volunteers. METHODS: The participants in this single-blind cross-over study (n = 6) were randomly assigned to receive one of three different formulations: plain-formulated aspirin (PF), extended-release aspirin that released acetylsalicylic acid steadily over 5 h (EX5) or an extended-release formulation that released 49% of the drug during the first 2 h after intake (EX2) and the rest of the dose during the subsequent 5 h. Laboratory analyses were done for platelet aggregation and thromboxane B2 (in whole blood), 6-keto-prostaglandin F1alpha (in leucocytes), neutrophil nitric oxide production and plasma nitrite/ nitrate levels. RESULTS: The PF and EX2 formulations inhibited platelet aggregation by 97% with no significant difference in effect between the two. In contrast, maximum inhibition of aggregation by the EX5 formulation was only 30%. Similar effects were found for platelet thromboxane production: PF and EX2 led to 99% inhibition, whereas EX5 led to 76% inhibition (P < 0.05). The inhibition of prostacyclin production differed in all three treatments (63% for PF, 40% for EX2 and 24% for EX5). The increase in leucocyte nitric oxide production also differed in all three treatments (1.01-fold the basal value with PF, 1.4-fold with EX5 and 3.6-fold with EX2). Both extended release formulations maintained high levels of nitric oxide production 24 h after the last dose, whereas in the PF period nitric oxide concentration had returned to basal values after this time. The changes in plasma nitrite concentrations in each period of treatment were similar to those seen for leucocyte nitric oxide. CONCLUSION: The pharmacodynamic profile of the extended-release formulations was better than that of plain-formulated aspirin in terms of thromboxane/prostacyclin balance and nitric oxide production. However, the EX2 formulation inhibited platelet function more effectively than did the EX5 formulation. PMID- 11871384 TI - Identifying general practice patients diagnosed with asthma and their exacerbation episodes from prescribing data. AB - OBJECTIVE: To determine the reliability of identifying patients diagnosed with asthma in general practice and their asthma exacerbation episodes from prescribing data. DATA SOURCE: Automated database from 17 general practitioners (29,805 patients) in the northern Netherlands. STUDY DESIGN: Sensitivity, specificity and predictive values of four criteria for identifying patients diagnosed with asthma and two criteria for identifying asthma exacerbation episodes were calculated using the registered diagnosis as gold standard. RESULTS: Prescription of one or more anti-asthma medications identified 95% of patients with an asthma diagnosis (positive predictive value 0.70), while two or more anti-asthma medications identified 71% (positive predictive value 0.79). A combination of oral corticosteroids or antibiotics identified 55% of exacerbations. CONCLUSIONS: Asthma patients can be identified reliably from prescribing data, but identification of asthma exacerbations was poor. The preference for one criterion over another for identifying patients diagnosed with asthma will depend on the reason for patient selection. PMID- 11871385 TI - Detection of cis-trans isomers of a synthetic peptide fragment of erythropoietin. AB - The synthesis of the protected fragment t-butoxycarbonyl-alanine-isoleucine serine(benzyl)-proline (Pro)-Pro-OH derived from the hormone erythropoietin is described. The analysis of the peptide by high-pressure liquid chromatography (HPLC) and thin-layer chromatography (TLC) yields apparently inconsistent results. Although HPLC consistently indicates the presence of only one component, TLC reveals a number of distinct species. Because satisfactory amino acid analysis and fast atom bombardment-mass spectrometry results are obtained, we think it possible that the distinct components arise from the cis-trans isomerization of the peptide bonds to the prolyl residues. An analysis using capillary electrophoresis under basic conditions identifies four components in the final product. Also, under similar conditions proton nuclear magnetic resonance spectroscopy is able to confirm the presence of cis and trans isomers. The results from this study demonstrate the usefulness of each of the four techniques in identifying the isomerism of the standard amino acid-Pro bond with respect to the peptide's ionic state. PMID- 11871387 TI - Different clinical value of minimal residual disease after autologous and allogenic stem cell transplantation for chronic lymphocytic leukemia. PMID- 11871386 TI - Simultaneous determination of caffeine, theobromine, and theophylline by high performance liquid chromatography. AB - This work relates the development of an analytical methodology to simultaneously determine three methylxanthines (caffeine, theobromine, and theophylline) in beverages and urine samples based on reversed-phase high-performance liquid chromatography. Separation is made with a Bondesil C18 column using methanol water-acetic acid or ethanol-water-acetic acid (20:75:5, v/v/v) as the mobile phase at 0.7 mL/min. Identification is made by absorbance detection at 273 nm. Under optimized conditions, the detection limit of the HPLC method is 0.1 pg/mL for all three methylxanthines. This method is applied to urine and to 25 different beverage samples, which included coffee, tea, chocolate, and coconut water. The concentration ranges determined in the beverages and urine are: < 0.1 pg/mL to 350 microg/mL and 3.21 microg/mL to 71.2 microg/mL for caffeine; < 0.1 pg/mL to 32 microg mL and < 0.1 pg/mL to 13.2 microg/mL for theobromine; < 0.1 pg/mL to 47 microg/mL and < 0.1 pg/mL to 66.3 microg/mL for theophylline. The method proposed in this study is rapid and suitable for the simultaneous quantitation of methylxanthines in beverages and human urine samples and requires no extraction step or derivatization. PMID- 11871389 TI - Why antiviral CD8 T lymphocytes fail to prevent progressive immunodeficiency in HIV-1 infection. PMID- 11871388 TI - Comprehensive cytogenetic and molecular genetic characterization of the TI-1 acute myeloid leukemia cell line reveals cross-contamination with K-562 cell line. PMID- 11871391 TI - Thr325IIe polymorphism of the TAFI gene does not influence the risk of myocardial infarction. PMID- 11871390 TI - Management of anticoagulation-associated toxicity during large-volume leukapheresis of peripheral blood stem cell donors. PMID- 11871392 TI - Molecular andrology: advances in the new millennium. PMID- 11871393 TI - Investigation of trace element sources from an industrialized area (Aviles, northern Spain) using multivariate statistical methods. AB - In the industrialized urban area of Aviles (Spain), pooled soil samples from grassland sites were taken after careful sampling design and then investigated for their content of trace elements. A combination of statistic and geostatistic techniques proved to be a reliable tool in the interpretation of the analytical results. Geochemical groups and areas influenced mainly by anthropogenic activities were distinguished by means of classic multivariate methods (factor analysis and cluster analysis) and innovative geostatistical tools (Fourier spectral analysis). The constitution of the groups is supported by the linked results of methods sequentially distributed in order to extract as much information as possible from the original data. PMID- 11871394 TI - Arsenic poisoning in groundwater: health risk and geochemical sources in Bangladesh. AB - Of the 2508 water samples analyzed in 10 districts of Bangladesh, 51%, on an average, contained arsenic levels of 0.05 to 2.50 mg/l. 95% of nail, 96% of hair, and 94% of urine samples contained arsenic above the normal level. Approximately 3.58 million people out of a total of 17.92 million who are drinking water containing arsenic levels >0.20 mg/l are potentially exposed to high risk of health hazard. Eight thousand and five hundred arsenic patients are identified; they are suffering from various skin lesions, gangrene in leg, skin, lung, bladder, liver, and renal cancer. A big portion of the total population is highly vulnerable to various internal cancers. Lowest arsenic concentration in drinking water producing dermatological disease is found to be 0.103 mg/l. However, the exposure time to develop arsenicosis varies from case to case reflecting its dependence on arsenic level in drinking water and food, nutritional status, genetic variant of human being, and compounding factors. This study has determined the high intensity of fluorescent humic substances in drinking water containing elevated concentrations of arsenic and very low concentrations of heavy metals. The synergistic/antagonistic effect of fluorescent compounds present in drinking water may aggravate the toxicity of arsenic. Geochemical study suggests that arsenic may be released from both reductive dissolution of Fe and Mn (oxy)hydroxide and microbial oxidation of organic matter. PMID- 11871395 TI - Olfactory responses of Plutella xylostella natural enemies to host pheromone, larval frass, and green leaf cabbage volatiles. AB - The parasitoids Trichogramma chilonis (Hymenoptera: Trichogrammatidae) and Cotesia plutellae (Hymenoptera: Braconidae), and the predator Chrysoperla carnea (Neuroptera: Chrysopidae), are potential biological control agents for the diamondback moth, Plutella xylostella (Lepidoptera: Yponomeutidae). We present studies on the interactions between these bioagents and various host-associated volatiles using a Y olfactometer. T chilonis was attracted to a synthetic pheromone blend (Z11-16:Ald, Z11-16:Ac, and Z11-16:OH in a 1:1:0.01 ratio), to Z11-16:Ac alone, and to a 1:1 blend of Z11-16:Ac and Z11-16:Ald. C. plutellae responded to the blend and to Z11-16:Ac and Z11-16:Ald. Male and female C. carnea responded to the blend and to a 1:1 blend of the major components of the pheromone, although no response was elicited by single compounds. Among the four host larval frass volatiles tested (dipropyl disulfide, dimethyl disulfide, allyl isothiocyanate, and dimethyl trisulfide), only allyl isothiocyanate elicited significant responses in the parasitoids and predator, but C. plutellae and both sexes of C. carnea did respond to all four volatiles. Among the green leaf volatiles of cabbage (Brassica oleracea subsp. capitata), only Z3-6:Ac elicited significant responses from T. chilonis, C. plutellae, and C. carnea, but C. plutellae also responded to E2-6:Ald and Z3-6:OH. When these volatiles were blended with the pheromone, the responses were similar to those elicited by the pheromone alone, except for C. carnea males, which had an increased response. The effect of temperature on the response of the biological agents to a mixture of the pheromone blend and Z3-6:Ac was also studied. T. chilonis was attracted at temperatures of 25-35 degrees C, while C. plutellae and C. carnea responded optimally at 30-35 degrees C and 20-25 degrees C, respectively. These results indicate that the sex pheromone and larval frass volatiles from the diamondback moth, as well as volatile compounds from cabbage, may be used by these natural enemies to locate their diamondback moth host. PMID- 11871396 TI - A novel mode of ecdysozoan growth in Caenorhabditis elegans. AB - Whereas growth in many ecdysozoa is associated with only molting, larval growth in nematodes, specifically Caenorhabditis elegans, is thought to be continuous and exponential. However, this has never been closely investigated. Here we report several detailed studies of growth in wild-type and dwarf C. elegans strains. We find that apparent exponential growth between hatching and adulthood comprises a series of linear phases, one per larval stage, with the linear growth rate increasing at successive molts. Although most structures grow continuously, the buccal cavity does not; instead, it grows saltationally at molts, like arthropod structures. We speculate that these saltational changes in mouth size permit changes in growth rate and that molting exists in nematodes to facilitate rapid growth. We study the cellular basis of this growth in the hypodermis. At each larval stage, lateral seam cells produce daughters that fuse with hyp7, a syncytium covering most of the worm. We find that seam cells and fusing daughter cells obtain larger sizes in successive molts. The total seam cell volume remains constant relative to the size of the worm. However, fusing daughter cells contributes only a very small amount directly to hypodermal growth, suggesting that most hyp7 growth must be intrinsic. Thus, dwarfism mutations studied principally act via adult syncytial growth, with cell size being near normal in both dbl-1 and dpy-2 mutant worms. We speculate that the main function of seam cell proliferation may be to supply the hypodermis with additional genomes for the purpose of growth. PMID- 11871397 TI - Oral mucosa: variations from normalcy, part I. AB - Many lesions can be identified in the oral mucosa. Some are indicative of pathologic conditions restricted to the oral cavity, whereas others are signs of systemic disorders. A third group of alterations is not severe enough to be considered pathologic; however, knowledge of this group is mandatory for a correct differential diagnosis to be established. This is the first article in a 2-part series concerning all of these pseudopathologic conditions. PMID- 11871398 TI - Identification of the human P450 enzymes involved in the in vitro metabolism of the synthetic steroidal hormones Org 4060 and Org 30659. AB - 1. The type of human P450 enzymes involved in the in vitro metabolism of Org 4060 and Org 30659, two synthetic steroidal hormones currently under clinical development by NV Organon for use in oral contraceptive and hormone replacement therapy, was investigated. 2. Both steroids were mainly hydroxylated at the 6beta position in incubations with human liver microsomes. 3. The results from experiments with supersomes, correlation studies as well as inhibition studies with ketoconazole, a selective inhibitor of CYP3A, strongly suggest that the CYP3A family plays a significant role in the 6beta-hydroxylation of both steroids. 4. Measurements of kinetic parameters of P450 enzymes that could metabolize both steroids, combined with the fact that CYP3A4 is known to be the most abundant P450 enzyme in the human liver, indicate that CYP3A4 will be of major importance for the in vivo human metabolism of Org 4060 and Org 30659. PMID- 11871399 TI - Impact of induced aniseikonia on stereopsis with random-dot stereogram. AB - In this work, we evaluate the impact of induced aniseikonia on stereopsis. For this, we determined the disparity range (maximum disparity), a parameter related to the size of the physical region that can be perceived stereoscopically. A significant decline in the disparity range was detected with aniseikonia induced by size lenses of 3% for five of the seven observers tested; 5% was necessary for the other two observers. The data indicate the influence of aniseikonia in stereopsis and the need to minimize such impact. These results may be useful in surgical processes such as the correction of pseudophakic patients and refractive surgery in which aniseikonia can be induced to alter the binocular function of the patient. PMID- 11871400 TI - The effect of ocular dominance on visual field testing. AB - PURPOSE: During standard automated perimetry (SAP), some patients experience visual disturbances in the tested eye while the other eye is covered with an opaque occluder. It is possible that a binocular interaction producing an inhibitory response in the nonoccluded eye, such as rivalry or Ganzfeld blankout, may be the causative factor, particularly when the dominant eye is occluded. The objective of this experiment was to determine whether subjective visual disturbances occurring during conventional perimetric test conditions were related to ocular dominance and to investigate the effect of these disturbances on measurements made during threshold visual field analysis. METHOD: Ocular dominance was determined by questioning and objective testing on 55 normal subjects. Each subject underwent program 24-2 Full Threshold SAP on a Humphrey Field Analyzer, and an opaque black patch was used to occlude the nontested eye. After testing, patients were asked to report symptoms of visual disturbance characteristic of rivalry or blankout, and the relationship between ocular dominance and visual disturbances was investigated. To determine whether symptoms of rivalry or blankout had affected visual field quantification, comparisons of short-term fluctuation, mean deviation, and false-negative errors were performed between eyes with and without visual disturbances. RESULTS: A total of 24 of 55 subjects reported visual disturbances consistent with rivalry or blankout (44%). Sixteen subjects complained of the phenomenon in one eye, and eight complained of the phenomenon in both eyes. Of the 16 experiencing disturbances in one eye only, nine cases occurred during occlusion of the dominant eye. The association between ocular dominance and visual disturbances was not found to be significant (p > 0.10). No significant differences in short-term fluctuation (p = 0.78), mean deviation (p = 0.64), or false-negative errors (p = 0.10) were found between eyes with and without visual disturbances. CONCLUSIONS: Patients undergoing standard automated perimetry with opaque patch occlusion of the nontested eye often experience visual disturbances consistent with rivalry or blankout, although these disturbances do not cause increased within-test variability or reduced sensitivity as quantified by visual field global indices. In terms of summary visual field indices, ocular dominance does not appear to affect visual field test results. PMID- 11871401 TI - The effect of handpiece spray patterns on cutting efficiency. AB - BACKGROUND: High-speed handpieces' spray ports direct coolant at the cutting interface. The authors evaluated the effect of the number of ports and their positions on cutting rates, or CRs. METHODS: The authors performed cutting studies on a machinable ceramic block using an established testing regimen. One port, three-port and four-port handpieces from one manufacturer were operated at maximum torque and rotation speed under a water flow of 25 milliliters per minute. The authors made 6-millimeter long edge and groove cuts in 13-mm cross section blocks using six medium-grit diamond burs for each handpiece. Each bur cut a total of 78 mm. The authors determined CR as the time to transect the block and analyzed the data by two-way analysis of variance with post hoc Scheffe tests. RESULTS: CRs varied by the type of cut and the number of spray ports. No differences were found in CRs for the three handpieces during edge cutting. The one-port handpiece cut significantly slower (P < .001) than did the three- and four-port handpieces during groove cutting. CONCLUSION: The data indicate that the number of handpiece spray ports, and their positioning relative to the bur affect water supply to the cutting interface and, consequently, the CR under these study conditions. CLINICAL IMPLICATIONS: Optimal cutting efficiency requires good coolant access, especially within restricted areas. A multiple-port handpiece may be advantageous when preparing the interproximal region for a crown or a proximal box, owing to the better water spray pattern. Dentists should consider the influence of the number of spray ports when selecting handpieces for cutting procedures. PMID- 11871402 TI - A gift to be used sparingly. PMID- 11871403 TI - Yellow fever: an update. AB - Yellow fever, the original viral haemorrhagic fever, was one of the most feared lethal diseases before the development of an effective vaccine. Today the disease still affects as many as 200,000 persons annually in tropical regions of Africa and South America, and poses a significant hazard to unvaccinated travellers to these areas. Yellow fever is transmitted in a cycle involving monkeys and mosquitoes, but human beings can also serve as the viraemic host for mosquito infection. Recent increases in the density and distribution of the urban mosquito vector, Aedes aegypti, as well as the rise in air travel increase the risk of introduction and spread of yellow fever to North and Central America, the Caribbean and Asia. Here I review the clinical features of the disease, its pathogenesis and pathophysiology. The disease mechanisms are poorly understood and have not been the subject of modern clinical research. Since there is no specific treatment, and management of patients with the disease is extremely problematic, the emphasis is on preventative vaccination. As a zoonosis, yellow fever cannot be eradicated, but reduction of the human disease burden is achievable through routine childhood vaccination in endemic countries, with a low cost for the benefits obtained. The biological characteristics, safety, and efficacy of live attenuated, yellow fever 17D vaccine are reviewed. New applications of yellow fever 17D virus as a vector for foreign genes hold considerable promise as a means of developing new vaccines against other viruses, and possibly against cancers. PMID- 11871404 TI - United Nations rallies support to tackle HIV/AIDS. PMID- 11871405 TI - An unusual cause of mild urticaria. PMID- 11871406 TI - Prospects for better tuberculosis vaccines. AB - Tuberculosis remains one of the top three infectious disease killers. Treatment is long and expensive and drug resistant strains of Mycobacterium tuberculosis are already on the rise. The current vaccine, BCG, is ineffective in parts of the world where the disease is most widespread and therefore the search for a novel, more effective vaccine is paramount. In this review we discuss the current state of vaccine research, including the identification of candidate antigens and the current methods used for their evaluation. PMID- 11871407 TI - Infectious origins of, and molecular mimicry in, Guillain-Barre and Fisher syndromes. AB - Guillain-Barre syndrome (GBS), characterised by limb weakness and areflexia, is the prototype of postinfectious autoimmune diseases, and Campylobacter jejuni is the most frequent antecedent pathogen. GBS subsequent to C jejuni enteritis is associated with a severe, pure motor axonal variant and IgG antibodies against GM1, GM1b, GD1a, or GalNAc-GDla, gangliosides expressed in human peripheral nerves. Lipopolysaccharides of C jejuni isolated from GBS patients have ganglioside-like epitopes. Cytomegalovirus is the most common viral antecedent infection. Patients with demyelinating GBS who have had a recent CMV infection have severe sensory deficits and anti-GM2 IgM antibody. CMV-infected fibroblasts express the GM2 epitope. Fisher syndrome (FS), characterised by ophthalmoplegia, ataxia, and areflexia, is a GBS variant associated with anti-GQ1b IgG antibody. GQ1b is enriched in the cranial nerves that innervate the extraocular muscles. Some patients develop FS after C jejuni infection, and the lipopolysaccharide present bears the GQ1b epitope. Molecular mimicry is a possible cause of GBS and FS. PMID- 11871408 TI - Hepatitis C as an opportunistic infection in HIV patients. PMID- 11871409 TI - Can antibiotic-resistant nosocomial infections be controlled? AB - Three decades ago infection-control programmes were created to control antibiotic resistant nosocomial infections, but numbers of these infections have continued to increase, leading many to question whether control is feasible. Meticillin resistant Staphylococcus aureus and vancomycin-resistant enterococci were major problems during the 1990s. Many hospitals have tried antibiotic control but with limited efficacy against these pathogens. Studies of antibiotic restriction, substitution, and cycling have been promising, but more definitive data are needed. Increased compliance with hand hygiene would help but is unlikely to control this problem alone as a result of frequent contamination of other surfaces even when hands are cleansed and high transmission rates when hand hygiene is neglected. For 17 years, the Centers for Disease Control and Prevention have recommended contact precautions for preventing nosocomial spread of important antibiotic-resistant pathogens. Many studies confirm that this approach works when sufficient active-surveillance cultures are undertaken to detect the reservoir for spread. However, most healthcare facilities have not yet tried this approach. PMID- 11871410 TI - Sharing needles may produce artificial leishmaniasis cycle. PMID- 11871411 TI - Borna disease virus infection in psychiatric patients: are we on the right track? AB - Animals infected with Borna disease virus (BDV) typically present with neurological dysfunction including behavioural abnormalities. Seroepidemiological surveys suggested that BDV infection can occur in human beings and is associated with mental disorders. Partly contradictory results from studies employing RT-PCR and serological screening led to debate over whether BDV can infect people at all. Critical evaluation of available data led to doubts about the diagnostic value of RT-PCR-based test results. A more consistent picture has emerged from serological studies because seropositive cases were found more frequently among psychiatric patients than among normal controls, supporting the notion that BDV might indeed be responsible for some psychiatric disorders. This view is now challenged by the observation that human BDV-reactive antibodies are of low avidity and might therefore represent cross-reacting antibodies. It remains to be shown whether these antibodies are indeed induced by BDV or by related antigens of unknown identity. PMID- 11871412 TI - Molecular genetics helps to probe the history of malaria. PMID- 11871413 TI - President of Kenya proposes sex ban. PMID- 11871415 TI - WHA urges more action on infectious diseases. PMID- 11871414 TI - Planned interruptions of anti-HIV treatment. AB - Highly active antiretroviral therapies (HAART) lower morbidity and mortality of HIV infection, but are unable to eradicate HIV and may cause side-effects. Planned interruptions of HAART are studied for three reasons: (1) to stimulate the anti-HIV immune response after viraemia has been suppressed by treatment; (2) to increase time off drug, to improve quality of life and diminish side effects and costs; and (3), among individuals whose virus has become resistant to treatment, to induce reversion of resistance to wild-type and therefore to improve the chances of success of subsequent salvage therapy. Regarding immune stimulation, the most promising results come from rare patients who started HAART during primary HIV infection. Up to 60% controlled viraemia below 5,000 copies/mL for up to 1 year without HAART. Among the majority of patients who start HAART later, during chronic HIV infection, preliminary results suggest that after repeated cycles on and off therapy, about 20% remain with a viraemia below 5,000 copies/mL after 12 weeks' treatment interruption. For the 80% who rebound to higher viraemia, additional immune-stimulatory manoeuvres are being considered such as administration of cytokines, or specific immune treatment by anti-HIV vaccines. When treatment is interrupted in patients who fail HAART and who have drug-resistant HIV, the drug-sensitive wild-type replaces the resistant quasispecies in 30-60% of patients. It remains to be seen whether this phenomenon increases the chances of successful salvage therapy. The risks and benefits of treatment interruption still have to be compared in large clinical trials, which are currently in the planning stage. PMID- 11871416 TI - Germany steps up research on the vascular endothelium. PMID- 11871417 TI - S Ragnar Norrby--Director General of the Swedish Institute for Infectious Disease Control. Interview by Pam Das. PMID- 11871418 TI - Looking out for wildlife. PMID- 11871419 TI - Infectious diseases surveillance update. PMID- 11871420 TI - Can we beat MRSA now we know its genome sequence? PMID- 11871421 TI - Was the education I provided effective? PMID- 11871422 TI - Glucose control in in-patients. PMID- 11871423 TI - A treatment for Mr WP: thiazolidinediones after troglitazone. PMID- 11871424 TI - Effectiveness of a hospital diabetes specialist nursing service. PMID- 11871425 TI - Abstracts of the 102nd Annual Meeting of the American Roentgen Ray Society. Atlanta, Georgia, USA. April 28-May 3, 2002. PMID- 11871426 TI - Cancer risk by occupation and socioeconomic group among men--a study by the Association of Swiss Cancer Registries. PMID- 11871427 TI - [Textual research on personnels revising Xin xiu ben cao (newly Revised Herbology)]. AB - The problem is investigated by studying annals of Tang - Song period and relevant materials. It is claimed that at the time when Sujing's report was submitted in the second year of Xianqing reign, the officials surveilling and revising the text include in order: ZHANG SUN Wuji, LI Ji, YU Zhining etc, together with SU Jing, the total number being 23. Changes appeared in this number at the time when the work was finished in the fourth year of Xianqing reign. Though, by then, the number involved was still 23, yet the order of LI Ji and ZHANG SUN Wuji was reversed, and Xin Mao jiang took the place of YU Zhining. In addition, the birth time of SU Jing was investigated to be 595 - 600. PMID- 11871428 TI - [The life and early achievements of Quan Shaoqing, a modern hygienist]. AB - After graduation from the Peiyang Medical College in Tianjin, Quan Shaoqing went to Tibet, introducing modern medicine. At the same time introducing Tibet to the outside world in the form of photos. He was a medical professor of Tientsin Medical college in early time. He assisted Dr. Wu Lieneh to prevent the epidemic of plague in three northeastern provinces in 1910 ~ 1911. Later, he was in charge of the eradication of plague in Inner Mongolia and Shanxi in 1918 with distinguished achievements. As an influential modern Chinese hygienist, he attended international symposium on medicine hygiene. PMID- 11871429 TI - [Research on origin and development of "Zhang Yi Tie", a celebrated physician of Xin'an]. AB - The "Xin' an Medicine", beginning from Jin - Tang and flourished in Ming - Qing periods, occupied a significant position in the history of TCM. A detailed textual and research on the origin and development of "Zhang Yi Tie" of Dingtan, She Country, Huizhou, a representative medical family of Xin' an clinical physician, is made here. Maintaining that the title of "Zhang Yi Tie" began in Jiajing reign of the Ming dynasty, was applied until now. In the period of over 450 years of 15 generations successive ages, Zhang was conversant with acute febrile diseases, intractable miscellaneous disease of internal medicine, with the "powdered medicine" inherited. The clinical characteristics of all generation of Zhang were explored, holding that regional medical schools, representative medical families are one of the important constructive forms in the developmental process of TCM. PMID- 11871430 TI - [A brief introduction of achievements in the study of the art of history of Shanghan (Cold Injury)]. AB - The achievement is described in five subheadings, viz. relevant people, relevant works, relevant events, academic relationship, and characteristic of stages. It is held that, the early development is not enough to demonstrate its significance and historical position, even less to show its development direction. It is essential to make further investigation. PMID- 11871431 TI - [Progress of pediatrical physio - pathological theory in modern TCM]. AB - Progress was made in significance of pediatrical physio pathology in modern TCM, including the elaboration on the role of dry dampness as etiological factor in pediatrics; the elaboration of renovation of theory of fetal - toxicity from an eugenic viewpoint; the popularization of variolation and its immunological mechanism, and its verification conclusion in TCM; combination of traditional wind syndrome - like western physiopathology in cerebral - neurology. PMID- 11871432 TI - [The meridian flowing and ancient Chinese philosophy]. AB - The theories of Yin - Yang and Five - Evolutive phase were extensively applied after pre - Qin and both Han dynasties. In the Song - Ming dynasties, the background of New Confucianism, the achievements of art of Yi(Changes) advocated by the "School of Configuration - Arithmatic" were applied, in which the method of "Adopting of Jia" was almost entirely derived from the above school, mixing the theories of heavenly stems - earthly branches, Five - evolutive phases and Yin - Yang as a whole. An ancient system of Chronological treatment was thus established creatively by integrating clinical experience together. The system is a tight system of grafting medicine and philosophy formed on the basis of successfully utilizing the philosophical framework, thinking proposed by ancient physicians with keen observation and deep ponderation. The system is, by then, an advanced one, which is also an inevitable one. PMID- 11871433 TI - [The conscious of mutual infiltration of nature of things (Cmnn) and alchemy and swallowing stone]. AB - By Cmnn, it refers to an idea that a matter can possess the nature of another matter completely irrelevant to the former one. This idea consistently played an important role in the pursuing of selection of longevity medicines in ancient time. Through this idea, the alchemists connected the life activity of the human body to the natural materials like gold and cinnabar to reinforce their belief of longevity. PMID- 11871434 TI - [Study on development history of Taoist medicine in Wudang]. AB - This accomplishment of textual research on the development of Wudang Taoist medicine covers the history from the record on Wudang priests practicing medicine in early period, herb pills and internal alchemy in Song - Yuan period to Taoist medicine when at its peak and the strive of doctors of Taoist priests made a vitalize Taoism after its period of declination. PMID- 11871435 TI - [A brief history of the several missionary hospitals in the modern Anhui province]. AB - In the modern Anhui province, the west missionary established clinics and hospitals to 74 in Anhui. There were four hospitals in those had been longer time and larger scale and more entirely manage system. It was larger influence for the formation and development of the modern Anhui western medicine. In the courses, the missionary - doctors promoted the spreading of the west medicine in the modern Anhui, but large quantities of Chinese doctors and nurses made with the activity contributes for these. PMID- 11871436 TI - [Introduction to management of environmental sanitation in public concession in Shanghai in 1930s]. AB - The environmental sanitation in Shanghai public concession is one of the regions with better administration. A complete set of administration rules was proposed by the Hygienic Department and Logistic Department of its public Security Bureau. It also spent a wealth of money to organize personnel and material supply to treat its environmental sanitation. This experience is worthy of summarization. PMID- 11871437 TI - [Fujian oversea Chinese and Chinese traditional pharmacist in Southeast Asia]. AB - Hakka from Yongding and southern part of Fujian, engaged in Chinese traditional medicine enterprise in south - east Asia, make up the majority of overseas from Fujian. They played an important part in the development of traditional Chinese medicine and insurance of people's health, in planting Chinese herbs, opening pharmaceutical factory, organizing pharmaceutical company and associations of medicine etc. They made great contributions to the communication and cooperation of medicine between PR China and other countries, especially Fujian and south - east Asia. PMID- 11871438 TI - [On the differences and similarities of pulse - taking between TCM and Tibetan medicine]. AB - There is a basis for the comparison of pulse - taking between TCM and Tibetan Medicine. There are connections and differences as well. The history of TCM pulsology is time - honored. There were contacts between TCM and Tibetan Medicine before the founding of Tubo dynasty. Many TCM classics from inland China were brought into Tibet by Princess Whencheng. Clear influence of Chinese pulsology can be seen in Tibetan pulsology. However, Tibetan pulsology has its own uniqueness. By combining imported knowledge with the experience of Tibetan people themselves, Tibetan pulsology is thus well developed. PMID- 11871440 TI - JAMA patient page. Hypertension. PMID- 11871439 TI - [The influence of conception of time - space - orientation in pre - Qin society on traditional Chinese medicine theories]. AB - The article discusses the views of the time - space - orientation of Wuxing (Five - Evolutive Phase), Yin - yang and Sifeng (Four - Wind), and its influence on traditional Chinese medical theories. It is shown that traditional Chinese medical theories are developed and established on the basis of the science and technology and social thought of the pre - Qin period, with some contents to be further discussed, and bias would be resulted if the issue is interpreted by concepts of post - Qin period. PMID- 11871441 TI - From the Centers for Disease Control and Prevention. Pertussis--United States, 1997-2000. PMID- 11871442 TI - From the Centers for Disease Control and Prevention. Progress toward elimination of perinatal HIV infection--Michigan, 1993-2000. PMID- 11871443 TI - From the Centers for Disease Control and Prevention. Hypothermia-related deaths- Utah, 2000, and United States, 1979-1998. PMID- 11871444 TI - [2nd Annual Meeting of the French Society of Neurology. Paris, France, 10 January 2002. Abstracts]. PMID- 11871446 TI - Evaluation of light transmission and distribution materials for Lunar and Martian bioregenerative life support. AB - The materials that were selected and evaluated in this study in the context of bioregenerative advanced life support included polymer optical cables, for transmission of photosynthetic photon flux (PPF), and light pipe, woven optical pad and light-emitting fiber (LEF) for PPF distribution. All materials exhibited significant fidelity in transmitting the spectral characteristics of the artificial lluminator's Xenon-Metal Halide lamp. The PPF attenuation values for the polymer cables EL-200, EL-300, EL-400, and EL-500 were not significantly distinguishable from one another nor from that of the fused-silica cable of 0.34 dB/m. With the exception of EL-100 and EL-700, which had significantly lower PPF transmission efficiencies of 54.9%/m and 66.6%/m, respectively, all the other polymer cables had PPF transmission efficiencies of over 85%/m which, except for EL-300, were not significantly different from one another nor from that of the fused-silica cable of 93.2%/m. The highest PPF output efficiency achieved for the 7.1-cm light pipe 14.7%, for its maximum pipe length of 100 cm. At a constant pipe length of 50 cm, the PPF output efficiency of the 10-cm light pipe of 0.71% was significantly lower than that of the 7.1-cm light pipe of 10.54%. The PPF output for the woven optical pad was determined to be 36.3%. The PPF output efficiency for the LEF without the optic fastener was determined to be 27.1%, whereas that for the LEF with the optic fastener was 50.3%, that is, the maximum value of PPF output efficiency in the study. The polymer optical cables, light pipe, woven optical pad, and LEF exhibited significant regularity and symmetry in their PPF output spatial distributions. PMID- 11871445 TI - Azolla-Anabaena symbionts and microbial mat as nitrogen-fixing biocatalysts for bioregenerative space life support. AB - This study was conducted to characterize the responses of selected nitrogen fixing biocatalysts to various environmental parameters that are likely to be encountered in a scaled-up nitrogen-fixing bioregenerative unit for use in a biologically based life support system (BLSS). The results showed that while both Azolla filiculoides and Azolla nilotica thrived on standard nitrogen-free liquid media, only Azolla filiculoides maintained its growth rate in the presence of 50% by volume of aerobic-bioreactor potato effluent (ABPE) in its liquid medium. The growth rate and Anabaena cavity population of Azolla filiculoides also remained undiminished in the presence of as much as 10 mM of NO3- in its liquid medium. The tolerance of Azolla filiculoides for NO3- however, lay somewhere between 10 and 20 mM of NO3-. Azolla filiculoides failed to grow at 5 mM or greater concentrations of NH4+. The growth rate of the Oscillatoria-dominated microbial mat was significantly elevated by increased silage loading density (16 g/L) as well as by adding 5 mM of NO3- into its liquid medium. Neither the growth rate of Azolla filiculoides nor that of the microbial mat was affected by three levels of photosynthetic photon flux (PPF) (520, 265, and 125 micromoles m-2 s-1) under high-pressure sodium or cool fluorescent lamps. PMID- 11871447 TI - Characterizations of high-intensity red and blue light-emitting diodes (LEDs) as a light source for plant growth. AB - Recently developed high-intensity red and blue light-emitting diodes (LEDs), which constitute a potentially improved light source for controlled-environment plant growth applications such as in vitro micropropagation and biologically based advanced life support (ALS) for space missions, were characterized in this study. Blue 2 LED and Red 1 LED consistently yielded the highest and lowest voltage drop readings, respectively, for all the electrical current settings tested (5-50 mA), with Blue 1 LED producing voltage drops falling nearly in the middle of the readings for the first two LEDs. At the standard current setting of 20 mA, Blue 2 LED and Blue 1 LED required 2.5 and 1.6 times more electrical power, respectively, than did Red 1 LED. At the standard current of 20 mA, the average photosynthetic photon flux (PPF) for Red 1 LED, Blue 1 LED, and Blue 2 LED were 180, 145, and 36 micromoles m-2 s-1, respectively. Red 1 LED peaked at 460 micromoles m-2 s-1 at 50 mA, Blue 1 LED at 200 micromoles m-2 s-1 at 40 mA, and Blue 2 LED at 40 micromoles m-2 s-1 at about 25 mA. For all current settings, the electrical conversion efficiency of Red 1 was approximately two times greater than that of Blue 1 LED. The electrical conversion efficiency of Blue 1 and of Red 1 LED peaked in between 10 and 20 mA, at about 13 mA for Blue 1 LED and at about 15 mA for Red 1 LED. The normalized PPF distributions for both Red 1 LED and Blue 1 LED were independent of the various magnitudes of electrical current (20, 30, 40, and 50 mA) that were applied to the LEDs. PMID- 11871448 TI - An evaluation of the vapor phase catalytic ammonia removal process for use in a Mars transit vehicle. AB - This article describes the design specification of the Vapor Phase Catalytic Ammonia Removal (VPCAR) process and the relative benefits of its utilization in a Mars Transit Vehicle application. The VPCAR process is a wastewater treatment technology that combines distillation with high-temperature catalytic oxidation of volatile impurities such as ammonia and organic compounds. PMID- 11871449 TI - Advantages of Sabatier for extended duration manned missions. AB - As manned space missions become longer and go farther away (i.e., Mars missions), the cost of resupply missions becomes substantial and even impractical. In order to reduce the logistics penalty for air revitalization in manned spacecraft, breathing oxygen (O2) must be recovered from metabolic carbon dioxide (CO2). The Sabatier CO2 reduction system is a key component of an integrated air revitalization system. The heart of the Sabatier system is the chemical catalyst bed that reacts carbon dioxide with hydrogen to form methane and water. Product water from a Sabatier subsystem would positively affect the current International Space Station (ISS) water balance and Mars missions would also benefit from the use of product methane as a propellant. This article focuses on the potential benefits of using the Sabatier subsystem for ISS and potential Mars mission applications. PMID- 11871450 TI - Space health requirements: the challenges. AB - This article explores the application of theoretical knowledge to clinical situations based on general systems theory and space health requirements to familiarize health care providers with requirements for the space environment. Preparation for extended periods of humans living in the space environment requires carefully planned delivery system that will promote and maintain health. Past, present, and future efforts for the establishment of'space health delivery systems are discussed. The National Aeronautics and Space Administration (NASA), Man-systems integration standards, NASA-STD-3000 Volume 1-MSIS, Revision B (1995, July), Houston, TX, National Aeronautics and Space Administration documents will be reviewed. Health care services will be supported by the available crew health care and emergency services systems. Providing health care in the extreme space environment with limited resources in which to carry out health practices offers challenges to health care providers. PMID- 11871451 TI - Telehealth: important concepts for future nursing practice in space environments. AB - The concept of telehealth has become a vital issue to healthcare providers in this day of instantaneous, varied, and technology-based communications. This article discusses the issues and implications of telehealth to nurses on Earth and in future space environments. Telehealth will be defined as currently implemented and the legal ramifications of practice across state lines, national borders, rural, and in remote and hazardous locations (space environments: orbital, Moon and Mars bases) will be delineated. The age of information is now here, and the age of communication is beginning. Telehealth is and will be an important means of providing communication links and healthcare to clients and providers alike. Healthcare professionals need to be aware of important concepts related to telehealth for their current and future practice. PMID- 11871452 TI - Distant oceanic paramedical practice: an example of environmental health nursing in remote hazardous locations. AB - Occupational health practitioners, whether they be medical practitioners or nursing practitioners, work in an ever-increasing diversified environment--from banks and insurance companies and other commercial outlets in the tertiary field to light and heavy manufacturing and engineering--but even this specialized area of practice has various subspecialities that are "off the beaten track." This article will focus on one of these fields--that of the petrochemical industry. Oil and gas production can be found both on land and at sea and this article looks at some of the different types of units from healthcare perspective. The method of practicing as an offshore nurse is described. A summary of practical tasks and skills is also presented within the setting of an offshore medic's job description. This is based on the current practices and legislation that is in force at this time, not only within the UK but also including Europe as a whole. The reader will attain a general understanding of how this industry operates. However, more relevant to the readership, a comprehensive idea of the medical and par-medical knowledge and skills required to practice in a worldwide context as an offshore medic/nurse is presented. Some of the items discussed include the qualifications needed, the tasks undertaken (medical and nonmedical), and some basic study of the medical problems that are encountered while working within marine and land-based hostile environments. PMID- 11871453 TI - Introduction: human performance in extreme environments. PMID- 11871454 TI - Operational psychology countermeasures during the Lunar-Mars Life Support Test Project. AB - The Crew and Thermal Systems Division at the NASA Johnson Space Center conducted a series of human-rated tests, termed the Lunar-Mars Life Support Test Project (LMLSTP), designed to advance technology in closed life support systems. As the duration of these tests lengthened, the psychological factors associated with placing humans in these environments became increasingly salient to successful mission completion. A number of psychological activities were conducted to ensure successful operations and protect crew member well-being, including individual crew member selection, crew composition, training and preparation, family inclusion, educational briefings, in-mission tracking, operational interventions, and postmission repatriation. This article describes these activities, the rationale behind their design, the similarities and differences to techniques utilized for spaceflight, and considerations related to designing psychological countermeasures for confined environments. In addition to testing physical and engineering systems, the LMLSTP series functioned as an effective testbed for developing operational concepts and countermeasures for extended space missions. PMID- 11871455 TI - Teaming humans and automated systems in safely engineered environments. AB - Human factors research has historically been concerned with interactions among humans, machines, and the environments in which they operate. The introduction of intelligent, automated systems to support human performance and safety in a variety of challenging environments, including undersea systems, high-performance aircraft, and orbiting space stations, has introduced a new dynamic to these relationships. Many missions cannot be accomplished without the active participation of both intelligent human and intelligent machine members. This is particularly true in environments where individuals operate in isolation and without easy access to support crews. However, the teaming of these two systems that function in such different ways can lead to new types of error, with the human frequently unable to determine what the machine is doing and why. The challenge, then is to develop automated systems that support, rather than confound, the human user. Aerospace human factors research has been a leader in attempting to understand human-automation interactions and in establishing guidelines for the design and use of automated systems. This article discusses some of the human-automation interaction problems that have been observed operationally, what the existing research reveals, and several approaches that are being pursued to avoid "disconnects" between humans and automation. Once these disconnects are overcome, intelligent humans and intelligent machines will be able to work together more productively, thus leading to our furthered presence in a variety of challenging environments. PMID- 11871456 TI - The effects of personality and interpersonal relations on crew performance during space simulation studies. AB - There has been a substantial increase in the number of people living and operating in isolated, confined, and artificially engineered environments, such as spacecraft, deep diving, weather stations, submarines, and polar outposts. This article gives an overview of research undertaken in a variety of extreme environments in an effort to better understand how semiautonomous, task-oriented groups operating within these environments develop over time, as well as identification of the individual characteristics that promote performance under such circumstances. Research reviewed includes space simulation studies for the European Space Agency (ESA) where groups were isolated in hyperbaric chambers, as well as findings from polar expeditions, space missions, submarine missions, and other military settings. Findings from the space simulation studies in hyperbaric chambers provided empirical evidence for interpersonal issues anecdotally reported in Antarctica and in other isolated, operational team environments, such as "scapegoating" of deviant crew members, displacement of aggression to outside personnel, and time patterns in psychological reactions. No indications of a "psychological limit" for how long people can tolerate remaining in isolation and confinement were found. Certain personality characteristics were consistently associated with coping, and individuals characterized by strong achievement motivation combined with interpersonal sensitivity seemed to adapt better than others. Together, these results have implications for selection and training of people operating within extreme environments. PMID- 11871457 TI - Gender differences in the willingness to engage in risky behavior: a terror management perspective. AB - Two studies examined the effects of mortality salience inductions on men and women's willingness to engage in risk-taking behaviors. In Study 1, a sample of American college students (N= 101) were exposed to either a mortality salient or a control condition and then rated their willingness to engage in a variety of risk-taking activities. In Study 2, a sample of Israeli high-school students (N = 106) completed a self-esteem measure, were exposed to either a mortality salient or control condition, and then rated their willingness to use various psychoactive substances that were offered in three different hypothetical scenarios. In both studies, findings indicated that mortality salience led to higher willingness to engage in risky behaviors in men but not in women. Study 2 also revealed that self-esteem moderated the effect of mortality salience on the willingness to use psychoactive substances but only when they were offered by a friend. The discussion offers a terror management perspective of risk-taking behaviors. PMID- 11871458 TI - A phenomenographic approach to the meaning of death: a Chinese perspective. AB - The purpose of this study was to investigate qualitative and quantitative differences in Chinese children's concepts of death, as reflected in their drawings, and to analyze this conceptual development as it related to background variables (such as gender, age, religious belief, and heath status). Participants were 239 children in 6 grade groups recruited from primary and junior high school. The children were asked to draw their impression of the word "death" and to give a verbal commentary of what they had drawn. The drawings were analyzed according to a phenomenographic method and assigned to one of 3 superordinate and 12 subordinate qualitative categories, adapted from M. E.Tamm and A. Granqvist (1995). Metaphysical and biological death concepts dominated, while psychological death concepts were depicted least. Consistent with previous studies of the development of concepts of death in children, biological death concepts were most common for the younger age groups, and metaphysical death concepts were found predominately in the older age groups. Chi-square analysis revealed no significant differences among death concept categories as a function of the participants' gender, health status, religious belief, funeral attendance, or prior death of relatives or pets. The results are interpreted as providing a unique window on death concepts among Chinese children. PMID- 11871459 TI - The assessment of grief among Hong Kong Chinese: a preliminary report. AB - This is a preliminary report of a study that examined the clinical usefulness of the Chinese Grief Reaction Assessment Form (GRAF). The validity, reliability, and factor structure of the form were also explored. We administered the GRAF to 180 bereaved individuals during intake interview for bereavement counseling. The GRAF has obtained good internal Cronbach's alpha reliability of .89 and showed its validity in being able to discriminate between the grief reactions of bereaved individuals experiencing anticipated and unanticipated death in the predicted direction. Bereaved women had stronger grief reaction than men. The item "I do not want to abandon him/her" obtained the highest mean ranked scores among both bereaved men and bereaved women. Our findings support the use of the GRAF as a clinical tool to assess psychological symptoms associated with bereavement among Hong Kong Chinese, which also holds promise for research and program evaluation. PMID- 11871460 TI - The role of gender and suicide precipitant in attitudes toward nonfatal suicide behavior. AB - This study examined factors affecting young adults' attitudes about nonfatal suicidal behavior. It evaluated how respondent sex, respondent gender identity, the precipitant of the suicidal act (i.e., a relationship loss, an achievement failure, or a physical illness), and gender of the suicidal person influence reactions to a suicidal decision. In this study of nonfatal suicidal behavior, like in studies of suicide, attitudes were least negative when the suicidal act was in response to a physical illness. Men were more likely to agree with and accept the suicidal decision than women. Androgynous persons, on the other hand, tended to view the decision to kill oneself as foolish, independent of precipitant. They also reported less agreement, acceptance, and sympathy for such decision. The implications of these findings for the prevention of suicidal behaviors are considered. Because gender seems to play a role in the acceptability of suicidal behavior, prevention programs ought to explicitly examine gender issues in attitudes toward suicidal behavior. PMID- 11871461 TI - Effect of antimicrobial agents on the ecological balance of human microflora. AB - The normal microflora acts as a barrier against colonisation of potentially pathogenic microorganisms and against overgrowth of already present opportunistic microorganisms. Control of growth of opportunistic microorganisms is termed colonisation resistance. Administration of antimicrobial agents, therapeutically or as prophylaxis, causes disturbances in the ecological balance between the host and the normal microflora. Most studies on the impact of antimicrobial agents on normal microflora have been carried out on the intestinal flora. Less is known on the effects on oropharyngeal, skin, and vaginal microflora. Disturbances in the microflora depend on the properties of the agents as well as of the absorption, route of elimination, and possible enzymatic inactivation and/or binding to faecal material of the agents. The clinically most common disturbances in the intestinal microflora are diarrhoea and fungal infections that usually cease after the end of treatment. A well-balanced microflora prevents establishment of resistant microbial strains. By using antimicrobial agents that do not disturb colonisation resistance, the risk of emergence and spread of resistant strains between patients and dissemination of resistant determinants between microorganisms is reduced. In this article, the potential ecological effects of administration of antimicrobial agents on the intestinal, oropharyngeal, and vaginal microflora are summarised. The review is based on clinical studies published during the past 10 years. PMID- 11871462 TI - Pathogenesis of HIV-associated cardiovascular complications. AB - Reviews and studies published before the introduction of highly active antiretroviral therapy (HAART) have tracked the incidence and course of HIV infection in relation to cardiac illness in both children and adults. The introduction of HAART regimens has significantly modified the course of HIV disease, with longer survival rates and improvement of life quality in HIV infected people expected. However, early data raised concerns about HAART being associated with an increase in both peripheral and coronary arterial diseases. In this review we discuss HIV-associated cardiovascular complications focusing on pathogenetic mechanisms that could have a role in diagnosis, management, and therapy of these complications in the HAART era. PMID- 11871463 TI - Towards an internet civil defence against bioterrorism. AB - Approaches towards the public-health prevention of bioterrorism are too little, and too late. New information-based approaches could yield better homeland protection. An internet civil defence is presented where millions of eyes could help to identify suspected cases of bioterrorism, with the internet used to report, confirm, and prevent outbreaks. PMID- 11871464 TI - Mary Jane Cardosa--bringing local solutions to Sarawak, Malaysia. PMID- 11871465 TI - The mystery of Bernstein's cold. PMID- 11871466 TI - Where would global health be without charity? PMID- 11871467 TI - US withdraws writ against AIDS drug production in Brazil. PMID- 11871468 TI - Fighting bulls off the menu in Spain. PMID- 11871469 TI - Pneumoccocal vaccination programme begins in Australia. PMID- 11871470 TI - Trypanasome coat provides potential for new drugs. PMID- 11871471 TI - Global drug buying programme bolsters tuberculosis control. PMID- 11871472 TI - Cyclic peptides yield promising antimicrobial results. PMID- 11871474 TI - New candidate for malaria vaccine. PMID- 11871476 TI - Infectious diseases surveillance update. PMID- 11871473 TI - New rabies vaccine for India. PMID- 11871475 TI - Leptospira brings fresh challenge to adventure sports. PMID- 11871477 TI - Cholera strikes Afghanistan. PMID- 11871478 TI - If foot and mouth disease were a disease of human beings. PMID- 11871479 TI - Infections in the homeless. AB - Homeless people in developed countries have specific problems predisposing them to infectious diseases. Respiratory infections and outbreaks of tuberculosis and other aerosol transmitted infections have been reported. Homeless intravenous drug users are at an increased risk of contracting HIV, and hepatitis B and C infections. Skin problems are the main reason the homeless seek medical attention, and these commonly include scabies, pediculosis, tinea infections, and impetigo. Many foot disorders are more prevalent in the homeless including ulcers, cellulitis, erysipelas, and gas gangrene. The louse transmitted bacteria Bartonella quintana has recently been found to cause clinical conditions in the homeless such as urban trench fever, bacillary angiomatosis, endocarditis, and chronic afebrile bacteraemia. Treatment of homeless people is complicated by financial constraints, self-neglect, and lack of adherence. Patients with serious and contagious illnesses should be hospitalised. Physicians should be aware of these specific issues to enhance care. PMID- 11871481 TI - Clinical picture, Rat bite fever. PMID- 11871482 TI - Clinical and epidemiological aspects of Chagas disease. AB - Chagas disease is caused by the protozoan parasite Trypanosoma cruzi. During the past decades, after urban migrations, Chagas disease became frequent in cities and a health problem in non-endemic countries, where it can be transmitted vertically and by blood transfusion or organ transplantation. Microepidemics of acute Chagas disease have been reported, probably due to oral transmission. Heart involvement is the major feature of the disease because of its characteristics, frequency, and consequences, and is also the source of most controversies. The indeterminate clinical form, despite its good prognosis on at least a medium-term basis (5-10 years), has acquired increasing importance due to the controversial meaning of the abnormality of some tests and the myocardial focal lesions found in many patients. Simultaneous evaluation of the parasympathetic and of the sympathetic system in the heart has been done by spectral analysis of heart rate. The physiopathological and clinical significance of denervation in Chagas disease is still incompletely understood. There are major divergences of opinion on specific treatment during the chronic phase because of the doubts about cure rates. Changes of Chagas disease prevalence in many countries have been certified by the Pan American Health Organization, and are ascribed to large-scale vector control programmes with modern pyrethroid insecticides and to improvement in lifestyle. PMID- 11871483 TI - Time for India to act on AIDS. PMID- 11871480 TI - Efficacy of pneumococcal conjugate vaccines and their effect on carriage and antimicrobial resistance. AB - Pneumococcal conjugate vaccines have shown a high degree of success in preventing pneumococcal bacteraemia in children. They also reduce the acquisition of carriage of vaccine serotypes in the nasopharynx, and reduce otitis media caused by those serotypes. Non-vaccine serotypes, which can colonise vaccinated infants, are associated with otitis media in these children and lower the overall effectiveness of the vaccine to this disorder. Longer term studies, however, could show that immunised children develop immunity to a broad range of pneumococcal serotypes at a younger age than non-immunised children. Preliminary data suggest that these vaccines could reduce the burden of radiologically confirmed pneumonia. Pneumococcal conjugate vaccines interrupt the transmission of antibiotic-resistant pneumococci and thus decrease the burden of antibiotic resistance in immunised children and in their contacts. Studies are underway to assess conjugate vaccine efficacy against invasive disease, pneumonia, and all cause mortality in developing countries, and to assess the potential use of these vaccines in adults. PMID- 11871484 TI - Roche gives in to Brazil over AIDS drug. PMID- 11871485 TI - Cambodia launches Asian vaccine drive. PMID- 11871486 TI - WHO announce strategy to combat antimicrobial resistance. PMID- 11871487 TI - Viruses get the ride of their lives. PMID- 11871488 TI - Spain says good-bye to measles. PMID- 11871489 TI - West Nile virus in the USA--an update. PMID- 11871490 TI - Fluoroquinolones as pneumococcal therapy: closing the barn door before the horse escapes. PMID- 11871491 TI - Vancomycin-resistant Staphylococcus aureus: a new model of antibiotic resistance. AB - Vancomycin has been the most reliable therapeutic agent against infections caused by meticillin-resistant Staphylococcus aureus (MRSA). However, in 1996 the first MRSA to acquire resistance to vancomycin, was isolated from a Japanese patient. The patient had contracted a post-operative wound infection that was refractory to long-term vancomycin therapy. Subsequent isolation of several vancomycin resistant S. aureus (VRSA) strains from USA, France, Korea, South Africa, and Brazil has confirmed that emergence of vancomycin resistance in S aureus is a global issue. A certain group of S. aureus, designated hetero-VRSA, frequently generate VRSA upon exposure to vancomycin, and are associated with infections that are potentially refractory to vancomycin therapy. Presence of hetero-VRSA may be an important indicator of the insidious decline of the clinical effectiveness of vancomycin in the hospitals. Vancomycin resistance is acquired by mutation and thickening of cell wall due to accumulation of excess amounts of peptidoglycan. This seems to be a common resistance mechanism for all VRSA strains isolated in the world so far. PMID- 11871492 TI - Cationic peptides: effectors in innate immunity and novel antimicrobials. AB - Cationic antimicrobial peptides are produced by all organisms, from plants and insects to human beings, as a major part of their immediately effective, non specific defences against infections. With the increasing development of antibiotic resistance among key bacterial pathogens, there is an urgent need to discover novel classes of antibiotics. Therefore, cationic peptides are being developed through clinical trials as anti-infective agents. In addition to their ability to kill microbes, these peptides seem to have effector functions in innate immunity and can upregulate the expression of multiple genes in eukaryotic cells. One such function might involve the dampening of signalling by bacterial molecules such as lipopolysaccharide and lipoteichoic acid. PMID- 11871493 TI - Immunotherapy of sepsis. AB - Sepsis is a clinical syndrome that results from a systemic host response to an infection. The outcome of sepsis is poor, and mortality rates are as high as 30 40%. Sepsis is associated with the activation of multiple inflammatory pathways, including the cytokine network and the coagulation system. Sepsis can also result in an immunodepressed state that could leave patients more susceptible to secondary nosocomial infections. Modulation of the host response to infection has been studied as an adjunctive therapeutic approach in many preclinical investigations and clinical trials in the past 20 years. As a result of these studies our knowledge of the pathogenesis of sepsis has increased considerably. This review focuses on immunomodulatory strategies that have reached the phase of clinical evaluation in patients with sepsis. PMID- 11871494 TI - Systematic review and meta-analysis of antibiotic therapy for bone and joint infections. AB - We set out to evaluate the clinical efficacy of individual antibiotic agents for bone and joint infections in adults. Published and unpublished controlled trials reported between 1966 and 2000 were reviewed to determine if they involved random or quasi-random allocation to systemically administered antimicrobials or local antibiotic therapy for osteomyelitis and septic arthritis. Quiescence of infection after 1 year of follow-up was defined as the primary outcome measure. 22 trials containing 927 patients were eligible for final analysis. Varying proportions of the entire study population could be evaluated with respect to primary and secondary endpoints. Methodological quality was poor among most studies, and interpretability of results was further limited by small sample sizes, missing descriptions of patient populations and disease characteristics, and the frequent application of concomitant antibiotics. A trend towards improved, long-lasting infection control was observed in favour of a rifampicin ciprofloxacin combination versus ciprofloxacin monotherapy for the treatment of staphylococcal infections related to orthopaedic devices (absolute risk difference [ARD] 28-9%; 95% CI -0.7 to 54.4%). Obviously unbalanced comparative studies showed some benefit of ticarcillin for bone infections caused by Pseudomonas species. No significant differences in therapeutic efficacy were found among trials comparing oral fluoroquinolones with intravenous beta-lactam drugs for both end-of-treatment (OR 0.8; 0.5 to 1.4) and long-term results (OR 1.3; 0.8 to 2.1). A variety of drugs was used as controls, thereby leading to inconsistent findings of drug-related side effects. Only one randomised trial was suitable to investigate the impact of polymethylmethacrylate gentamicin bead chains compared with parenteral antibiotics for skeletal infections, although this study was biased by patients receiving both combined local and systemic antibiotic therapy. Whereas intention-to-treat evaluation suggested a therapeutic advantage of systemic over local therapy, this trend diminished in the per protocol analysis (1-year follow-up ARD -2.3;-17.5 to 10.8%). There exists little high-quality evidence on antibiotic therapy for osteomyelitis and septic arthritis. The observed heterogeneity among patient populations and medical and surgical treatment concepts preclude reliable inferences from the available data. PMID- 11871495 TI - Topical immunomodulators--progress towards treating inflammation, infection, and cancer. AB - Immunomodulators include both immunostimulatory and immunosuppressive agents. Only recently have the basic mechanisms of topical immunotherapy been elucidated. Besides topical contact sensitisers (eg, diphencyprone or dinitrochlorobenzene), newer agents of the imidazoquinoline family such as imiquimod and resiquimod act by inducing cytokine secretion from monocytes or macrophages (interferon-alpha, interleukin-12, tumour-necrosis factor-alpha). The locally generated immune milieu leads to a Th1-dominance and cell-mediated immunity that have been used clinically to treat viral infections such as human papillomavirus (HPV), herpes simplex virus (HSV), mollusca, and cancerous lesions including initial squamous cell and basal cell carcinoma in immunocompetent and immunosuppressed patients. While these agents improve antigen-presentation by dendritic cells, they also act on B cells and lead to the synthesis of antibodies such as IgG2a much like the recently discovered immunostimulatory CpG-sequences that stimulate innate immunity. These sequences act as "danger signals" since they occur in bacterial and viral DNA, but are selectively methylated and inactivated in the mammalian genome. They share the induction of the same cytokines as imidazoquinolines but they show different magnitudes and kinetics of response. Topical immunotherapy with immunostimulatory agents shows potential for effective and patient-friendly treatment of inflammatory, infectious, and cancerous skin diseases. Immunoenhancers such as imdazoquinolines and CpG-sequences also have adjuvant properties that could improve conventional (protein) and DNA vaccination against cancer, atopy, and allergies. PMID- 11871496 TI - Multidrug-resistant tuberculosis--can the tide be turned? AB - Despite continued control efforts, tuberculosis remains a leading cause of illness and death worldwide, with more cases today than at any previous time in history. Not only is there a global increase in the disease itself, there is a worrying rise in the number of cases resistant to the two principal antituberculosis drugs, isoniazid and rifampicin--so called multidrug resistance. PMID- 11871497 TI - A fatal sandwich. PMID- 11871498 TI - Kim Mulholland--drawing together the pieces of international child health. PMID- 11871499 TI - What's in a name? PMID- 11871500 TI - What good can come of this? PMID- 11871503 TI - Debate on essential medicines heats up. PMID- 11871501 TI - Infectious diseases threaten refugees entering Pakistan. PMID- 11871502 TI - European trial platform for AIDS, malaria, and tuberculosis. PMID- 11871504 TI - Biowarfare fears in India. PMID- 11871505 TI - HIV may underlie ALS-like condition. PMID- 11871506 TI - Artificial cows effectively combat tsetse flies. PMID- 11871507 TI - Transgenic mosquitoes to control malaria. PMID- 11871508 TI - Is protection in HIV infection due to Bw4 or not to Bw4? PMID- 11871509 TI - Male circumcision and HIV prevention: current knowledge and future research directions. AB - Over the past decade, numerous epidemiological studies have reported a significant association between lack of male circumcision and risk for HIV infection, leading to recommendations for male circumcision to be added to the armamentarium of effective HIV prevention strategies. We review the epidemiological data from studies that have investigated this association, including ecological, cross-sectional/case-control, and prospective studies. We discuss problematic issues in interpreting the epidemiological data, including the presence of other sexually transmitted infections, age of circumcision, and potential confounders such as religion, cultural practices, and genital hygiene. In addition, we review studies of biological mechanisms by which the presence of the foreskin may increase HIV susceptibility, data on risks associated with the circumcision procedure, and available data on the acceptability and feasibility of introducing male circumcision in societies where it is traditionally not practised. Although the evidence in support of male circumcision as an effective HIV prevention measure is compelling, residual confounding in observational studies cannot be excluded. Taken together with concerns over the potential disinhibiting effect of male circumcision on risk behaviour, and safety of the circumcision procedure, randomised trials of male circumcision to prevent HIV infection are recommended. An individual's choice to undergo male circumcision or a community's decision to promote the practice should be made in the light of the best available scientific evidence. More knowledge is required to assist individuals and communities in making those decisions. We conclude with recommendations for future research. PMID- 11871510 TI - Treatment of chronic hepatitis B. AB - This review updates the treatment of chronic hepatitis B infection. Complete eradication of hepatitis B virus (HBV) is not possible, so the efficacy of treatment has to be assessed by whether it can limit long-term cirrhosis-related complications. We discuss two major groups of treatments--immunomodulators (interferon alfa, thymosin alpha1, therapeutic vaccines) and nucleoside analogues (lamivudine, adefovir, entecavir, emtricitabine, beta-L-2'-deoxythymidine). To date, interferon alfa and lamivudine are the only two agents approved for chronic hepatitis B. Interferon alfa achieves a short-term outcome of around 20-30% loss of HBeAg. The efficacy is lower in Chinese patients, who are immunotolerant to HBV because of acquisition of the disease during early childhood, than in white patients. This difference is further confirmed on long-term follow-up. Interferon alfa does not affect the development of cirrhosis-related complications in Chinese patients, whereas in white patients, the frequency of long-term complications is reduced if interferon alfa is successful in inducing loss of HBeAg. Lamivudine profoundly suppresses viral replication and achieves an HBeAg seroconversion rate similar to that of interferon alfa. It is equally effective in Chinese and white patients because the main antiviral mechanism is through inhibition of reverse transcription of HBV during viral replication. However, long-term lamivudine therapy is associated with emergence of HBV variants, YMDD variants. Newer nucleoside analogues are being extensively investigated by studies in vivo and in vitro. Combination therapy with two or three nucleoside analogues or immunomodulators plus nucleoside analogues will be the future direction of treatment of chronic hepatitis B. PMID- 11871511 TI - Modern chemotherapeutic options for malaria. AB - Unlike HIV disease or tuberculosis, both of which are also major threats to public health throughout the tropics, uncomplicated malaria of whatever species can be cheaply and rapidly cured, usually in outpatients. However, in common with both HIV and tuberculosis, control of malaria is threatened by inadequate resources and by drug resistance. Africa carries the greatest burden of malaria mortality and morbidity; by no coincidence, Africa is also the most resource limited. The drugs for severe disease (quinine and the artemisinins) are largely unaffected by resistance so far, but the "first-line" drugs, mainly used by outpatients (eg, chloroquine and sulfadoxine-pyrimethamine), are a major cause for concern. Although effective drugs are available they are mostly too expensive for routine use. This article reviews the main drugs for malaria and outlines the therapeutic use of these drugs for uncomplicated and severe disease. The article then examines the challenges faced in the processes of changing policy, and the implementation of that policy shift. PMID- 11871512 TI - Control of multiply resistant cocci: do international comparisons help? AB - Antibiotic resistance has become a worldwide problem. However, the reasons for the uneven geographic distribution of antibiotic-resistant microorganisms are not fully understood. For instance, there are striking differences in the epidemiology of multiresistant gram-positive cocci between the USA and Germany. According to recent reports, the prevalence of high-level penicillin-resistant pneumococci (PRP), meticillin-resistant Staphylococcus aureus (MRSA), and vancomycin-resistant enterococci (VRE) in clinically relevant isolates of hospitalised patients in the USA and Germany are: PRP, 14% versus less than 1%; MRSA, 36% versus 15%; and VRE, 15% versus 1%. These disparities may be explained by several determinants: (1) diagnostic practice and laboratory recognition (all three pathogens); (2) clonal differences and pathogen transmissibility (VRE); (3) antibiotic prescribing practices (all three pathogens); (4) population characteristics, including extensive daycare exposure in the USA (PRP); (5) cultural factors (all three pathogens); (6) factors related to the health-care and legal system (all three pathogens); and (7) infection-control practices (MRSA and VRE). Understanding these determinants is important for preventing further spread of multiresistant cocci within the USA. A rational approach to national surveillance is urgently needed in Germany to preserve the favourable situation and decrease MRSA transmission. Finally, we suggest that a macro-level perspective on antibiotic resistance can broaden the understanding of this worldwide calamity, and help prevent further dissemination of multiply resistant microorganisms. PMID- 11871513 TI - Cryptosporidiosis: an update. AB - Cryptosporidiosis was recognised in human beings in 1976, and was prominent in the 1980s and 1990s as a cause of severe diarrhoeal illness in patients with AIDS. It is now additionally recognised as a major cause of waterborne diarrhoeal illness in developed regions, and as a pathogen with long-term effect on childhood growth and development in impoverished areas. This update focuses on recent changes in our understanding of the taxonomy of cryptosporidium, its epidemiology, effects, pathogenesis, diagnosis, and treatment. PMID- 11871514 TI - Mandatory screening and treatment of immigrants for latent tuberculosis in the USA: just restraint? AB - A report by the Institute of Medicine, Ending Neglect, and sponsored by the US Centers for Disease Control and Prevention, makes recommendations for achieving elimination of tuberculosis in the USA. Among them is the recommendation that a mandatory screening programme be introduced for latent tuberculosis infection in immigrants from high prevalence countries, and that the provision of a permanent residence card (green card) be linked to the completion of an approved course of preventive treatment. We examine the evidence put forward to support this proposal and assess whether such a mandatory programme for preventive treatment of individuals, who do not pose an immediate risk but could pose a risk in the future, meets internationally recognised standards for coercive public-health measures. We conclude from our analysis that there is reason to question (i) the risk analysis, (ii) the estimates of effectiveness of such a policy, (iii) the cost calculations, and (iv) the operational consequences put forward in the report. Moreover, we show that international standards for mandatory screening and treatment are not met. PMID- 11871515 TI - T Jacob John--taking India forward in infectious disease control. Interviewed by Pam Das. PMID- 11871516 TI - Something nasty in the grass. PMID- 11871517 TI - Ward grannies provide tender loving care. PMID- 11871518 TI - Clarification on pain management and nutrition in treating acute pancreatitis. PMID- 11871519 TI - Prone positioning device challenged. PMID- 11871522 TI - Mucormycosis: a case study. AB - Throughout the history of mucormycosis, from the first case in humans reported in 1885 by Paltauf, through publication by Gregory et al of the first observation of rhino-orbital cerebral mucormycosis in 1943, to the report by Harris in 1955 of the first known survivor, little has changed in the diagnosis and outcome of this disease. Although mucormycosis of any form--cerebral, cutaneous, rhinocerebral, intestinal, or pulmonary--is still a rarity, it should be suspected in patients who are diabetic or immunocompromised. Administration of amphotericin B, surgical debridement of infected tissue, correction of the underlying cause, and use of adjunctive HBO therapy remain the standard treatments. PMID- 11871523 TI - Prone positioning of patients with acute respiratory distress syndrome: applying research to practice. PMID- 11871524 TI - Nursing care after pneumonectomy in patients with invasive pulmonary aspergillosis. PMID- 11871525 TI - Use of blood cultures in critically ill patients. AB - Infection, bacteremia, and sepsis are frequent complications in critically ill patients. Ideally, the infectious agent is readily identified to facilitate timely treatment to promote the patient's recovery. Use of blood cultures is one method of identifying the pathogen. Fever is the primary indicator for obtaining blood samples for culture, but other indicators may be considered, depending on the patient's medical history and condition. Use of appropriate techniques when collecting blood samples for culture will decrease contamination and improve the likelihood of identification of the infectious agent. One new technique being tested for the identification of pathogens that cause bacteremia involves genetic technology and the polymerase chain reaction. The polymerase chain reaction is used to identify the DNA of bacteria that are present in the blood. Blood cultures may not always result in identification of the pathogen because the organism may not grow once placed in culture medium. This new method that uses the polymerase chain reaction may be more sensitive than blood cultures because it requires only DNA from bacteria. Although early studies have not been conclusive in terms of the benefits of this new technology, additional research will improve methods for identification of pathogens in critically ill patients. PMID- 11871526 TI - Preventing nosocomial pneumonia: evidence-based practice. PMID- 11871527 TI - Living-donor lobar lung transplantation: a case study. PMID- 11871528 TI - Traditional and nontraditional modes of mechanical ventilation. PMID- 11871529 TI - The Synergy Model. Overview of the theoretical review process. PMID- 11871530 TI - Description of the procedure for monitoring intra-abdominal pressure via an indwelling urinary catheter. PMID- 11871531 TI - Credentials can be an edge in critical care practice. PMID- 11871532 TI - Staffing shortages mean increased opportunities. PMID- 11871533 TI - Education paves way for career advancement. PMID- 11871534 TI - Temporary assignments can open many doors. PMID- 11871535 TI - Practice standards articulate expectations. PMID- 11871537 TI - The outlook is bright for critical care nurses. PMID- 11871538 TI - Seek ways to learn about critical care nursing. PMID- 11871539 TI - The resume is your first impression. PMID- 11871540 TI - Royal College of Nursing. Critical care nursing forum. Organ donation: opting out? PMID- 11871541 TI - Kinetics of nitrate, nitrite, and Cr(VI) reduction by iron metal. AB - The kinetics of nitrate, nitrite, and Cr(VI) reduction by three types of iron metal (Fe0) were studied in batch reactors for a range of Fe0 surface area concentrations and solution pH values (5.5-9.0). At pH 7.0, there was only a modest difference (2-4x) in first-order rate coefficients (k(obs)) for each contaminant among the three Fe0 types investigated (Fisher, Peerless, and Connelly). The k(obs) values at pH 7.0 for both nitrite and Cr(VI) reduction were first-order with respect to Fe0 surface area concentration, and average surface area normalized rate coefficients (kSA) of 9.0 x 10(-3) and 2.2 x 10(-1) L m(-2) h(-1) were determined for nitrite and Cr(VI), respectively. Unlike nitrite and Cr(VI), Fe0 surface area concentration had little effect on rates of nitrate reduction (with the exception of Connelly Fe0, which reduced nitrate at slower rates at higher Fe0 surface areas). The rates of nitrate, nitrite, and Cr(VI) reduction by Fisher Fe0 decreased with increasing pH with apparent reaction orders of 0.49 +/- 0.04 for nitrate, 0.61 +/- 0.02 for nitrite, and 0.72 +/- 0.07 for Cr(VI). Buffer type had minimal effects on reduction rates, indicating that pH was primarily responsible for the differences in rate. At high pH values, Cr(VI) reduction ceased after a short time period, and negligible nitrite reduction was observed over 48 h. PMID- 11871542 TI - Modeling the mass-action expression for bidentate adsorption. AB - The number of bidentate binding sites on a pristine surface (i.e., sites comprising two adjacent monodentate sites plus the space between them) can be substantially larger than the maximum number of bidentate molecules that can adsorb to the surface. When bidentate sorption occurs, the number of available bidentate sites decreases in response to direct occupation of some sites, but an even more significant loss results from the fact that several unoccupied sites immediately surrounding each adsorbed molecule can also become unavailable. Recognition of this phenomenon allows development of a model for the adsorption process that matches simulated data from Monte Carlo (MC) modeling extremely well. The model also explains the observation that, on a given surface with a given fractional occupation, the number of available bidentate sites depends on whether the occupied sites are populated by monodentate or bidentate adsorbed species. A model developed more than 60 years ago but not widely recognized by modern adsorption modelers also fits the MC simulations very well. The simulated data are also reasonably approximated by assuming that the number of available bidentate sites on a surface is proportional to the square of the number of available monodentate sites, although the fit is not as good as with the models mentioned above. By contrast, approximating the number of available sites as proportional to the number of monodentate sites to the first power yields predictions that do not match the simulations. The results have implications for modeling of both multidentate adsorption reactions and monovalent-divalent ion exchange. PMID- 11871543 TI - Mobilization of natural colloids from an iron oxide-coated sand aquifer: effect of pH and ionic strength. AB - Field and laboratory column experiments were performed to assess the effect of elevated pH and reduced ionic strength on the mobilization of natural colloids in a ferric oxyhydroxide-coated aquifer sediment. The field experiments were conducted as natural gradient injections of groundwater amended by sodium hydroxide additions. The laboratory experiments were conducted in columns of undisturbed, oriented sediments and disturbed, disoriented sediments. In the field, the breakthrough of released colloids coincided with the pH pulse breakthrough and lagged the bromide tracer breakthrough. The breakthrough behavior suggested that the progress of the elevated pH front controlled the transport of the mobilized colloids. In the laboratory, about twice as much colloid release occurred in the disturbed sediments as in the undisturbed sediments. The field and laboratory experiments both showed that the total mass of colloid release increased with increasing pH until the concurrent increase in ionic strength limited release. A decrease in ionic strength did not mobilize significant amounts of colloids in the field. The amount of colloids released normalized to the mass of the sediments was similar for the field and the undisturbed laboratory experiments. PMID- 11871544 TI - Modeling steady-state particle size spectra. AB - Fractal dimensions of marine aggregates are often estimated from the measured slopes of particle size spectra. One technique uses dimensional analysis to determine the dependence of the spectrum's slope with fractal dimension. In this paper, we use numerical simulations to examine the assumptions underlying the dimensional analysis approach to particle size spectra. We find that the slopes of numerically computed steady-state particle size spectra disagree with those predicted by dimensional analysis. The assumptions underlying the dimensional analysis approach that are responsible for the disagreement are as follows: only one coagulation mechanism operates in each particular particle size range, particle loss through sedimentation occurs at particle sizes larger than those for which differential sedimentation dominates, particles only interact with like sized particles. Including disaggregation steepens the slope of the particle size spectrum for both large and small particles and changes the shape of the spectrum. These results indicate that one should exercise caution when using the measured slopes of particle size spectra to estimate aggregate fractal dimension. PMID- 11871545 TI - Adsorption of Cu, Cd, and Ni on goethite in the presence of natural groundwater ligands. AB - The adsorption of copper, cadmium, and nickel on goethite was examined in natural groundwater samples from an infiltration site of the river Glatt at Glattfelden (Switzerland). Unfractionated dissolved organic matter was used at its natural concentrations. Metal concentrations were close to environmental conditions. Cu, Cd, and Ni presented the typical pH adsorption edge of cations. The major influence on metal adsorption was due to a strong organic ligand L(I), which inhibited adsorption of Cu, Cd, and Ni in the alkaline pH region. Complexation of Cu, Cd, and Ni by the natural organic ligands was described with a model defining a minimum number of discrete ligands: a strong ligand L(I) at low concentration and additional weaker ligands with higher concentrations. The adsorption of Cu, Cd, and Ni on the goethite surface in the presence of the natural organic ligands was adequately described by considering only surface complexation and complexation in solution by organic ligands. No ternary complexes had to be invoked in the model. The major effect was complexation by the strongest ligand, whereas interactions with other cations and anions had only a minor influence. Competition reactions between Cu and Ni for complexation with the same strong ligand L(I) were observed. PMID- 11871546 TI - Steady-state dissolution kinetics of aluminum-goethite in the presence of desferrioxamine-B and oxalate ligands. AB - This paper reports steady-state dissolution rates of synthetic low-substitution Al-goethites (mol % Al < 10) at pH 5 in the presence of the trihydroxamate siderophore, desferrioxamine B (DFO-B), and the common biological ligand, oxalate. The siderophore-promoted Fe release rate increased both with the level of Al substitution and with DFO-B concentration up to about 100 microM, after which a plateau occurred, suggesting a saturation effect from DFO-B adsorption as a precursor to dissolution. At concentrations above 200 microM, oxalate also enhanced the Fe release rate, which however was not influenced by Al substitution. For Al-goethites with mol % Al < 4, the Fe release rate in the presence of 40 microM DFO-B together with varying concentrations of oxalate was typically greater than the corresponding sum of dissolution rates in the presence of the two ligands alone. This synergism may be the combined result of the ability of oxalate to adsorb strongly at the goethite surface, thus promoting Fe release, and of the high selectivity of DFO for Fe(III). Ferric oxalate complexes formed during dissolution will likely lose Fe3+ by ligand substitution with DFO B, leading to the production of Fe(HDFO-B)+ and uncomplexed oxalate, the latter of which, in turn, could adsorb to the goethite surface again. For Al-goethites with mol % Al > 4, synergism was not apparent, which may signal the effect of a decreased surface density of Fe-OH sites associated with Al for Fe substitution. The oxalate-promoted release rates of Al did not scale with those of Fe, indicating incongruent dissolution. However, Al release rates in the presence of DFO-B did scale approximately with those of Fe but were not affected by the concentration of siderophore. These results are consistent with the presence of Al(OH)3 inclusions in Al-goethite. PMID- 11871548 TI - Measurement and dynamic modeling of trace metal mobilization in soils using DGT and DIFS. AB - The technique of diffusive gradients in thin-films (DGT) accumulates metals on a Chelex resin after their diffusive transport through a hydrogel. It lowers metal concentrations in soil solution adjacent to the device and induces resupply of metal associated with the solid phase. DGT devices were deployed in an alluvial gley soil for 21 different time periods between 4 h and 19.5 d. The accumulated masses of Cu, Cd, Ni, and Zn were used to calculate the distribution coefficient for labile metal, Kdl, and adsorption and desorption rate constants. Calculations were performed using a dynamic numerical model of DGT-induced fluxes in soils (DIFS). It assumes first-order exchange between solid phase and solution and diffusional transport in both the soil solution and the hydrogel. The DIFS model fitted changes in accumulated mass with time very well. Values of Kdl calculated from DIFS of 100 (Cd), 250 (Cu), 150 (Ni), and 150 (Zn) were larger than values of distribution coefficients estimated by exchange with Ca(NO3)2 but similar to those estimated by isotopic exchange (Cd and Zn only). These results suggest that the solid-phase pool of metal affected by the removal of labile metal by DGT, which operates on a time scale of minutes, is similar to the solid-phase pool of metal that can isotopically exchange with solution on a time scale of 2 d. Response times of minutes were consistent with interaction rates with surfaces, and desorption rate constants agreed with other reported values. An appraisal of the DIFS model demonstrated the importance of the labile pool size in the solid phase for controlling supply to a sink, such as DGT or a plant. As values of Kdl and kinetic parameters are obtained using DGT with minimal soil disturbance and by a similar mechanism to that involved in plant uptake, they may be pertinent to bioavailability studies. PMID- 11871547 TI - Influence of metal ion sorption on colloidal surface forces measured by atomic force microscopy. AB - Atomic force microscopy (AFM) is employed to directly measure colloidal surface forces between a silica particle and a smooth glass plate in an aqueous solution with or without the presence of copper ions. Without the presence of copper ions, results show that the force between these two surfaces is repulsive and that its magnitude decreases with increasing ionic strength and decreasing pH. The surface forces are calculated based on the Derjaguin-Landau-Verwey-Overbeek (DLVO) theory for constant surface charge and are then compared with AFM force measurements. A good agreement between theory and experimental data is reported except at very small separation distances (<3 nm) between the silica particle and the glass plate. This behavior may be attributed to non-DLVO forces, such as the hydration effect that results from the bounded water molecules on the surface of the silica particle, or to surface roughness. When copper ions are present in acidic aqueous solutions, the magnitude of the force is found to be the same as that without the presence of copper ions, which indicates that no sorption of copper ions by the silica particle occurs under these conditions. Near neutral pH, sorption of copper ions causes charge reversal for the silica particle from negative to positive. Therefore, the force between the silica particle and the glass plate changes from repulsive to attractive. The transient zeta-potential of the silica particle during sorption of copper ions is determined by representing the experimental data with the DLVO theory. In alkaline solutions, where removal of copper ions is known to occur mainly by bulk precipitation, the measured force is similar to that without the presence of copper ions, which suggests that sorption does not occur under such conditions. PMID- 11871549 TI - Simulation of the mobility of metal-EDTA complexes in groundwater: the influence of contaminant metals. AB - Reactive transport simulations were conducted to model chemical reactions between metal-EDTA (ethylenediaminetetraacetic acid) complexes during transport in a mildly acidic quartz-sand aquifer. Simulations were compared with the results of small-scale tracer tests wherein nickel-, zinc-, and calcium-EDTA complexes and free EDTA were injected into three distinct chemical zones of a plume of sewage contaminated groundwater. One zone had a large mass of adsorbed, sewage-derived zinc; one zone had a large mass of adsorbed manganese resulting from mildly reducing conditions created by the sewage plume; and one zone had significantly less adsorbed manganese and negligible zinc background. The chemical model assumed that the dissolution of iron(III) from metal-hydroxypolymer coatings on the aquifer sediments by the metal-EDTA complexes was kinetically restricted. All other reactions, including metal-EDTA complexation, zinc and manganese adsorption, and aluminum hydroxide dissolution were assumed to reach equilibrium on the time scale of transport; equilibrium constants were either taken from the literature or determined independently in the laboratory. A single iron(III) dissolution rate constant was used to fit the breakthrough curves observed in the zone with negligible zinc background. Simulation results agreed well with the experimental data in all three zones, which included temporal moments derived from breakthrough curves at different distances downgradient from the injections and spatial moments calculated from synoptic samplings conducted at different times. Results show that the tracer cloud was near equilibrium with respect to Fe in the sediment after 11 m of transport in the Zn-contaminated region but remained far from equilibrium in the other two zones. Sensitivity studies showed that the relative rate of iron(III) dissolution by the different metal-EDTA complexes was less important than the fact that these reactions are rate controlled. Results suggest that the published solubility for ferrihydrite reasonably approximates the Fe solubility of the hydroxypolymer coatings on the sediments. Aluminum may be somewhat more soluble than represented by the equilibrium constant for gibbsite, and its dissolution may be rate controlled when reacting with Ca-EDTA complexes. PMID- 11871550 TI - Reactions of hydroxyl radical with humic substances: bleaching, mineralization, and production of bioavailable carbon substrates. AB - In this study, we examine the role of the hydroxyl (OH*) radical as a mechanism for the photodecomposition of chromophoric dissolved organic matter (CDOM) in sunlit surface waters. Using gamma-radiolysis of water, OH* was generated in solutions of standard humic substances in quantities comparable to those produced on time scales of days in sunlit surface waters. The second-order rate coefficients of OH* reaction with Suwannee River fulvic (SRFA; 2.7 x 10(4) s(-1) (mg of C/L)(-1)) and humic acids (SRHA; 1.9 x 10(4) s(-1) (mg of C/L)(-1)) are comparable to those observed for DOM in natural water samples and DOM isolates from other sources but decrease slightly with increasing OH* doses. OH* reactions with humic substances produced dissolved inorganic carbon (DIC) with a high efficiency of approximately 0.3 mol of CO2/mol of OH*. This efficiency stayed approximately constant from early phases of oxidation until complete mineralization of the DOM. Production rates of low molecular weight (LMW) acids including acetic, formic, malonic, and oxalic acids by reaction of SRFA and SRHA with OH* were measured using HPLC. Ratios of production rates of these acids to rates of DIC production for SRHA and for SRFA were similar to those observed upon photolysis of natural water samples. Bioassays indicated that OH* reactions with humic substances do not result in measurable formation of bioavailable carbon substrates other than the LMW acids. Bleaching of humic chromophores by OH* was relatively slow. Our results indicate that OH* reactions with humic substances are not likely to contribute significantly to observed rates of DOM photomineralization and LMW acid production in sunlit waters. They are also not likely to be a significant mechanism of photobleaching except in waters with very high OH* photoformation rates. PMID- 11871551 TI - Effects of Fe(III) chemical speciation on dissimilatory Fe(III) reduction by Shewanella putrefaciens. AB - Shewanella putrefaciens, a heterotrophic member of the gamma-proteobacteria is capable of respiring anaerobically on Fe(III) as the sole terminal electron acceptor (TEA). Recent genetic and biochemical studies have indicated that anaerobic Fe(III) respiration by S. putrefaciens requires outer-membrane targeted secretion of respiration-linked Fe(III) reductases. Thus, the availability of Fe(III) to S. putrefaciens may be governed by equilibrium chemical speciation both in the solution phase and at the bacterial cell-aqueous or cell-mineral interface. In the present study, effects of Fe(III) speciation on rates of bacterial Fe(III) reduction have been systematically examined by cultivating S. putrefaciens anaerobically on a suite of Fe(III)-organic complexes as the sole TEA. The suite of Fe(III)-organic complexes spans the range of stability constants normally encountered in natural water systems and includes Fe(III) complexed to citrate, 5-sulfosalicylate, NTA, salicylate, tiron, and EDTA. Rates of bacterial Fe(III) reduction in the presence of dissolved chelating agents correlate with the thermodynamic stability constants of the Fe(III)-organic complexes, implying that chemical speciation governs Fe(III) bioavailability. Equilibrium Fe(III) sorption experiments measured the reversible coordination of Fe(III) with S. putrefaciens as a function of cell/Fe(III) concentration, time, and activity of competing chelating agents. Results show that S. putrefaciens readily sorbs dissolved Fe(III) but that adsorption is restricted by the presence of strong Fe(III)-chelating agents. Our results indicate that dissimilatory Fe(III) reduction by S. putrefaciens is controlled by equilibrium competition for Fe(III) between dissolved organic ligands and strongly sorbing functional groups on the cell surface. PMID- 11871553 TI - Collision frequencies of microbial aggregates with small particles by differential sedimentation. AB - Collision and coagulation rates between microbial aggregates and small particles were measured for individual aggregates (1.0-2.5 mm) that settled through a suspension of fluorescent yellow-green (YG) particles (2.83 microm) placed in a settling column. The microbial aggregates, with an average fractal dimension of 2.26, were generated in a lab-scale sequencing batch reactor (SBR) and also collected from a full-scale activated sludge (AS) treatment system. As calculated from comparisons between the settling velocities observed and those predicted by Stokes' law for impermeable particles, the average fluid collection efficiencies were 0.08 for the SBR aggregates and 0.14 for the AS flocs, which were much lower than those previously reported for nonbiological aggregates of latex microspheres. The collision frequency functions between microbial aggregates and small YG particles were 2 orders of magnitude lower than predicted by the rectilinear model but 1 order of magnitude greater than predicted by a curvilinear model. The overall scavenging efficiencies of suspended particles by the falling microbial aggregates compared well with those observed for the nonbiological aggregates, while the particle removal efficiencies from the flow internal to the microbial aggregates were 1 order of magnitude higher than those of the nonbiological aggregates. It is argued that the permeability of microbial aggregates could be reduced by exopolymeric material clogging the pores within the aggregates. The internal permeation through a bioaggregate thus may not be significant enough to be included in the calculation of its settling velocity; however, the intra-aggregate flow cannot be simply neglected where coagulation is concerned. Streamlines still can penetrate the interior of microbial aggregates, allowing greater collision frequencies with other particles than predicted by the curvilinear model. The narrow and convoluted internal flow passages resulting from the collection of extracellular polymeric substances may also contribute to the higher interior particle removal efficiencies of microbial aggregates than those of more permeable, nonbiological aggregates. PMID- 11871552 TI - Deposition and fate of arsenic in iron- and arsenic-enriched reservoir sediments. AB - Deposition of arsenic to the sediments of Haiwee Reservoir (Olancha, CA) has dramatically increased since March 1996 as a result of an interim strategy for arsenic management in the Los Angeles Aqueduct (LAA) water supply. Ferric chloride and cationic polymer are introduced into the Aqueduct at the Cottonwood treatment plant, 27 km north of the Haiwee Reservoir. This treatment decreases the average arsenic concentration from 25 microg/L above Cottonwood to 8.3 microg/L below Haiwee. Iron- and arsenic-rich flocculated solids are removed by deposition to the reservoir sediments. Analysis of sediments shows a pronounced signature of this deposition with elevated sediment concentrations of iron, arsenic, and manganese relative to a control site. Sediment concentrations of these elements remain elevated throughout the core length sampled (ca. 4% iron and 600 and 200 microg/g of manganese and arsenic, respectively, on a dry weight basis). A pore water profile revealed a strong redox gradient in the sediment. Manganese in the pore waters increased below 5 cm; iron and arsenic increased below 10 cm and were strongly correlated, consistent with reductive dissolution of iron oxyhydroxides and concurrent release of associated arsenic to solution. X ray absorption near-edge spectroscopy revealed inorganic As(V) present only in the uppermost sediment (0-2.5 cm) in addition to inorganic As(III). In the deeper sediments (to 44 cm), only oxygen-coordinated As(III) was detected. Analysis of the extended X-ray absorption fine structure spectrum indicates that the As(III) at depth remains associated with iron oxyhydroxide. We hypothesize that this phase persists in the recently deposited sediment despite reducing conditions due to slow dissolution kinetics. PMID- 11871554 TI - Aqueous copper sulfide clusters as intermediates during copper sulfide formation. AB - Using a combination of experimental techniques, we show that Cu(II) reduction by sulfide to Cu(I) occurs in solution prior to precipitation. EPR and 63Cu NMR data show that reduction to Cu(l) occurs during the reaction of equimolar amounts of Cu(II) with sulfide. 63Cu solution NMR data show that Cu(I) is soluble when bound to sulfide and is in a site of high symmetry. EPR data confirm that Cu(I) forms in solution and that the mineral covellite, CuS, contains only Cu(I). Mass spectrometry data from covellite as well as laboratory prepared solid and solution CuS materials indicate that Cu3S3 six-membered rings form in solution. These trinuclear Cu rings are the basic building blocks for aqueous CuS molecular clusters, which lead to CuS precipitation. In controlled titration experiments where sulfide is slowly added to Cu(II), Cu3S3 rings and tetranuclear Cu molecular clusters (Cu4S5, and Cu4S6) form; the rings are composed primarily of Cu(II). During cluster formation from Cu3S3 condensation, some Cu(II) is released back into solution, indicating that Cu(II) reduction does not occur until after Cu-S bond and higher order cluster formation. Analysis of the frontier molecular orbitals for Cu(II) and sulfide indicate that an outer-sphere electron transfer is symmetry forbidden. These results are consistent with the formation of CuS bonds prior to electron transfer, which occurs via an inner-sphere process. PMID- 11871555 TI - Carbon tetrachloride transformation in a model iron-reducing culture: relative kinetics of biotic and abiotic reactions. AB - Contributions of biotic (cell-mediated) and abiotic (mineral-mediated) reactions to carbon tetrachloride (CT) transformation were studied in a model iron-reducing system that used hydrous ferric oxide (HFO) as the electron acceptor, acetate as the substrate, and Geobacter metallireducens as a representative dissimilative iron-reducing bacteria (DIRB). Over a period of 2-3 weeks, nanoscale magnetite particles, Fe3O4, were consistently formed as a product of iron respiration in this system. CT transformation rates were measured independently in resting cell suspensions of G. metallireducens or in suspensions of washed magnetite particles recovered from spent cultures. Protein and surface area-normalized expressions were derived for the biotic and abiotic reaction rates, respectively. Using the yield of total protein and magnetite surface area formed during growth in the model system as a basis for comparison, the mineral-mediated (abiotic) reaction was estimated to be 60-260-fold faster than the biotic reaction throughout the incubation period. We conclude that G. metallireducens induces CT transformation in this system primarily through the formation of reactive mineral surfaces rather than via co-metabolic mechanisms. The findings indicate that reactive biogenic minerals could play a significant role in the natural attenuation of chlorinated solvents in iron-reducing environments. A novel approach for stimulating reductive transformation of contaminants may be to enhance the formation of reactive biogenic minerals in situ. PMID- 11871556 TI - P, As, Sb, Mo, and other elements in sedimentary Fe/Mn layers of Lake Baikal. AB - Distinct layers with accumulated iron and manganese oxyhydroxides are found in the recent sediments of Lake Baikal (Siberia). In the South and Central Basins, these concretions accumulate close to the sediment-water interface. In northern Lake Baikal and the area of Academician Ridge, however, massive Fe/Mn crusts are formed within several thousand years at redox fronts 10 to 15 cm below the sediment surface. In some places, precipitated iron and manganese oxyhydroxides are spatially separated. The patterns are a result of secondary iron and manganese oxide precipitation. This natural long-term experiment allows the analysis of competitive adsorption and coprecipitation of trace elements with iron and manganese oxides in sediments. Background concentrations in the sediment of oxoanions (P, As, Sb, Mo); of trace metals (Cr, V, Cu, Zn, Cd, Pb); and of Mg, Ca, Sr, La, Ce, Pr, Nd, and Sm were analyzed by inductively coupled plasma mass spectrometry. Despite the differences in catchment geology of the many tributaries, they are remarkably uniform in sediment cores from different basins of Lake Baikal. Enrichment factors of P and As within Fe crusts revealed concentrations up to 14 and 58 times higher than the background, respectively. No enrichment of P and As was found in the Mn layers. By contrast, Mo accumulated exclusively in the Mn layer with up to 35-fold enrichment. Sb was only slightly enriched in both the Fe and the Mn layers. Among the trace metals studied, only Cd was found at elevated concentrations with a preference for the Mn layer. Ca and Sr were correlated with both Fe and Mn accumulations. The study quantifies the well-known specific adsorption and coprecipitation of P and As at authigenic iron oxides and of Mo on manganese oxides. In addition, the enrichment of Cd at manganese oxides in contrast to the conservative behavior of Zn and Pb reveals highly selective accumulation processes. PMID- 11871557 TI - Effect of oxide formation mechanisms on lead adsorption by biogenic manganese (hydr)oxides, iron (hydr)oxides, and their mixtures. AB - The effects of iron and manganese (hydr)oxide formation processes on the trace metal adsorption properties of these metal (hydr)oxides and their mixtures was investigated by measuring lead adsorption by iron and manganese (hydr)oxides prepared by a variety of methods. Amorphous iron (hydr)oxide formed by fast precipitation at pH 7.5 exhibited greater Pb adsorption (gamma(max) = 50 mmol of Pb/mol of Fe at pH 6.0) than iron (hydr)oxide formed by slow, diffusion controlled oxidation of Fe(II) at pH 4.5-7.0 or goethite. Biogenic manganese(III/IV) (hydr)oxide prepared by enzymatic oxidation of Mn(II) by the bacterium Leptothrix discophora SS-1 adsorbed five times more Pb (per mole of Mn) than an abiotic manganese (hydr)oxide prepared by oxidation of Mn(II) with permanganate, and 500-5000 times more Pb than pyrolusite oxides (betaMnO2). X-ray crystallography indicated that biogenic manganese (hydr)oxide and iron (hydr)oxide were predominantly amorphous or poorly crystalline and their X-ray diffraction patterns were not significantly affected by the presence of the other (hydr)oxide during formation. When iron and manganese (hydr)oxides were mixed after formation, or for Mn biologically oxidized with iron(III) (hydr)oxide present, observed Pb adsorption was similar to that expected for the mixture based on Langmuir parameters for the individual (hydr)oxides. These results indicate that interactions in iron/manganese (hydr)oxide mixtures related to the formation process and sequence of formation such as site masking, alterations in specific surface area, or changes in crystalline structure either did not occur or had a negligible effect on Pb adsorption by the mixtures. PMID- 11871558 TI - Surface chemistry and dissolution kinetics of divalent metal carbonates. AB - A surface complexation model (SCM) for divalent metal carbonates (Ca, Mg, Sr, Ba, Mn, Fe, Co, Ni, Zn, Cd, and Pb) is developed based on new electrophoretic measurements and correlation between aqueous and surface reactions stability constants. This SCM postulates the formation of the following surface species: >CO3H0, >CO3-, >CO3Me+, >MeOH0, >MeO-, >MeOH2+, >MeHCO30, and MeCO3- within the framework of a constant capacitance of the electric double layer. It can be used to describe the surface-controlled dissolution kinetics of divalent metal carbonates and allows determination of the order of dissolution reactions with respect to rate-controlling protonated carbonate surface groups in acid solutions (>CO3H0) and hydrated metal groups (>MeOH2+) in neutral to alkaline solutions. The reaction order with respect to protonated carbonate groups increases from 2 for MnCO3 and ZnCO3 to 4 for NiCO3, whereas for hydrated surface metals, it augments from 2 for ZnCO3 to approximately 4 for MnCO3 and NiCO3. The dissolution rates at 5 < or = pH < or = 8 increase in the order Ni < Mg < Co < Fe < Mn < Zn < Cd < Sr < or = Ca approximately = Ba approximately = Pb and correlate nicely with water exchange rates from the aqueous solution into the hydration sphere of the corresponding dissolved cations. Such a correlation allows the generation for all carbonates of a model describing their dissolution/precipitation kinetics, including the effect of various ligands, provided that rate constants and their activation volumes for water exchange around Me(II)-ligand dissolved complexes are available. PMID- 11871559 TI - Kinetic model for Fe(II) oxidation in seawater in the absence and presence of natural organic matter. AB - A detailed kinetic model has been developed to describe the oxidation of Fe(II) in seawater in both the absence and the presence of natural organic material. Experimental data were collected using a luminol chemiluminescence-based method to measure Fe(II), assuming that both the inorganic and the organically complexed species were detected. In the absence of organic matter, the data were modeled based on the Haber-Weiss mechanism with the inclusion of a back-reaction of Fe(III) with superoxide and precipitation of Fe(OH)3. Both reactions were found to be significant using sensitivity analysis. When organic matter is present, the model was extended by organic complexation of Fe(II) and Fe(III) with the creation of a parallel oxidation pathway for Fe(II). Fe(II) oxidation at natural (nanomolar) concentrations was accurately predicted for a range of organic concentrations. The model also accounted for scavenging of superoxide by sub nanomolar levels of dissolved copper and by organic matter when present. The presence of a relatively strong Fe(III) binding ligand was observed to significantly increase the rate of Fe(II) oxidation, while ultimately retaining most of the iron in the system in dissolved (organically complexed) form. The complexation reactions and reaction of inorganic and organically bound Fe(II) with oxygen were found to be critical reactions in the system, while Fe(III) hydrolysis became unimportant even at low organic concentrations. The superoxide radical was also observed to have a major role in the cycling of iron due to its ability to act as both an oxidant and a reductant. The model indicates that the rate constant for the reaction of Fe(II) with O2 has generally been underestimated in previous work and that the secondary oxidation of Fe(II) by H2O2 and subsequently OH* plays a relatively minor role in these systems. PMID- 11871560 TI - Is silica really an anomalous oxide? Surface acidity and aqueous hydrolysis revisited. AB - The single-site Solvation, Bond Strength, and Electrostatic (SBE) model accounts for the anomalous position of silica onthe surface acidity versus aqueous acidity correlation developed for metal oxides, by considering the solvation energy change in the protonation reaction implemented through the dielectric constant (1/epsilon(k)) and the electrostatic energy change through the Pauling bond strength to bond length ratio (s/r) of the oxide. I address here why inclusion of the solid's dielectric constant brings silica into the same correlation as other oxides like TiO2, Al2O3, and Fe2O3. The solvation and electrostatic contributions are interpreted in terms of classical concepts such as chemical hardness, polarizability, ionicity, electronegativity, and local charge densities. Silica is acidic (PZC < 7), not because of its small dielectric constant, its tetrahedral coordination, or its high bond strength alone. Surface acidity depends largely on high values of the s/r ratio. The dielectric constant of the solid affects acidity mainly by reflecting the nature of water-surface interactions. Solids with large values of epsilon(k) are interpreted as being less polarizable and more ionic so that water, a hard polar solvent, interacts favorably with such surfaces and scales similar to water-water interactions regardless of whether the metal-oxide bond is in the solid or in the aqueous state. For these oxides, pKa(s) = pKa(aq) +/- 1. Silica, with a small dielectric constant, is interpreted as being more polarizable and more covalent so that water-SiO2 interactions scale differently than for the more ionic oxides. Such an interpretation when combined with the Partial Charge Model for metal hydrolysis suggests that the surfaces of RuO2, W03, Sb2O5, and Ta2O5 should be acidic similar to silica. But, unlike silica, they would lie on the pKa correlation defined by the other oxides because of their larger dielectric constants. The mixed oxide, AlPO4, is predicted to behave like silica. PMID- 11871561 TI - Redox processes controlling manganese fate and transport in a mountain stream. AB - The biogeochemical processes controlling the speciation and transport of manganese in a Colorado mountain stream were studied using a conservative tracer approach combined with laboratory experiments. The study stream, Lake Fork Creek, receives manganese-rich inflows from a wetland contaminated by acid mine drainage. A conservative tracer experiment was conducted on a 1300-m reach of the stream. Results indicate that manganese was progressively removed from the stream, resulting in a loss of 64 +/- 17 micromol day(-1) m(-1). Laboratory experiments using streamwater, mine effluent, and rocks from the stream showed the importance of surface-catalyzed oxidation and photoreduction on the speciation of manganese. The field and modeling results indicate that light generally promotes oxidation and removal of manganese from the stream, presumably through a photosynthetically enhanced oxidation process. Differences in Mn speciation within the stream suggest that reductive processes affect Mn speciation within the water column. These results identify the rapid oxidation and precipitation of MnOx as a dominant process within this freshwater stream. PMID- 11871562 TI - Phytoplankton-mediated redox cycle of iron in the epilimnion of Lake Kinneret. AB - The biological-mediated redox cycle of Fe was studied in the epilimnion of Lake Kinneret (Sea of Galilee), a mesotrophic lake in Israel. Multi-annual lake water sampling and incubation experiments were carried out to study Fe(III) reduction by natural phytoplankton populations and their possible role in inhibiting Fe(II) oxidation. The reduction characteristics of the dinoflagellate Peridinium gatunense, the dominant lake alga, were further examined in the laboratory. The steady-state concentration of Fe(II) calculated from the assessed reduction and oxidation rates was compared with Fe(II) measured in the lake in order to evaluate the significance of these processes to the lake Fe redox cycle. Nanomolar concentrations of Fe(II) were measured in the oxygenated, high pH, upper water layer of the lake throughout the year. Reduction rates of Fe by natural phytoplankton assemblages ranged between 0.1 and 10 nM/h. The highest reduction rates, determined in dinoflagellate-dominated lake waters, coincided with the highest concentrations of Fe(II) measured simultaneously in the lake. Iron(II) oxidation rates calculated from the measured lake Fe(II) and the obtained reduction rates were significantly slower than published abiotic Fe(II) oxidation rates. Indeed, Fe(II) oxidation rates measured in algal-enriched lake water were 30-fold slowerthan Fe(II) oxidation rates in natural water, demonstrating the potential for Fe(II) stabilization by the lake phytoplankton. PMID- 11871563 TI - Evidence for a dynamic cycle between Mn and Co in the water column of a stratified lake. AB - The geochemical behavior of Co in aquatic systems has often been related to the presence of Fe and Mn particles. A few studies have shown that Co is exclusively associated with particulate Mn, but the dynamics of Co and Mn cycling have never been determined in real time under natural conditions. In this study, we used a combination of analytical techniques to study the temporal and spatial evolution of Mn microparticles (MnOx) over 2 weeks in the water column of a shallow stratified lake (Paul Lake, MI). We report a temporal accumulation of dissolved Mn at the oxic-anoxic transition, and we show that this accumulation is due to the reductive dissolution of Mn particles. The reductant has not been identified, but abiotic reduction by sigmaH2S and ferrous iron is excluded because they are produced below the zone of MnOx reduction. Hybridization of RNA isolated from Paul Lake with oligonucleotide probes targeting the delta proteobacteria, which include metal-reducing species, suggests that their activity is greatest at and just below the oxic-anoxic transition, so that Mn reduction may be influenced by bacterial activity. Mn-oxidizing bacteria were isolated from this zone as well. We also demonstrate that the dynamic evolution of MnOx has a direct influence on the distribution of Co in the water column of this lake: dissolved Co is released during the reductive dissolution of MnOx and accumulates at the redox interface. PMID- 11871564 TI - Particle-scale understanding of the bioavailability of PAHs in sediment. AB - This study reports results of sediment bioslurry treatment and earthworm bioaccumulation for polycyclic aromatic hydrocarbon (PAH) contaminants found in sediment dredged from Milwaukee Harbor. A significant finding was that bioslurry treatment reduced PAHs on the sediment clay/silt fraction but not on the sediment coal-derived fraction and that PAH reduction in the clay/silt fraction correlated with substantial reduction in earthworm PAH bioaccumulation. These findings are used to infer PAH bioavailability from characterization of particle-scale PAH distribution, association, and binding among the principal particle fractions in the sediment. The results are consistent with work showing that the sediment comprised two principal particle classes for PAHs, coal-derived and clay/silt, each having much different PAH levels, release rates, and desorption activation energies. PAH sorption on coal-derived particles is associated with minimal biodegradation, slow release rates, and high desorption activation energies, while PAH sorption on clay/silt particles is associated with significant potential biodegradability, relatively fast release rates, and lower desorption activation energies. These characteristics are attributed to fundamental differences in the organic matter to which the PAHs are sorbed. Although the majority of the PAHs are found preferentially on coal-derived particles, the PAHs on the clay/silt sediment fraction are more mobile and available, and thus potentially of greater concern. This study demonstrates that a suite of tests comprising both bioassays and particle-scale investigations provide a basis to assess larger-scale phenomena of biotreatment of PAH-impacted sediments and bioavailability and potential toxicity of PAH contaminants in sediments. Improved understanding of contaminant bioavailability aids decision-making on the effectiveness of biotreatment of PAH-impacted sediments and the likelihood for possible reuse of dredged sediments as reclaimed soil or fill. PMID- 11871565 TI - Modeling metal removal onto natural particles formed during mixing of acid rock drainage with ambient surface water. AB - Studies have examined partitioning of trace metals onto natural particles to better understand the fate and transport of trace metals in the environment, but few studies have compared model predictions with field results. We evaluate the application of an empirical modeling approach, using surface complexation parameters available in the literature, to complex natural systems. In this work, the equilibrium speciation computer program PHREEQC was used along with the diffuse double-layer surface complexation model to simulate metal removal onto natural oxide particles formed during the mixing of acid rock drainage with ambient surface water. End-member solutions sampled in the Coeur d'Alene (CdA) Mining District in September 1999 from the Bunker Hill Mine and the South Fork Coeur d'Alene (SFCdA) River were filtered and mixed in several ratios. Solution chemistry was determined for end-members and mixed solutions, and X-ray diffraction (XRD) was used to determine the mineralogy of precipitate phases. Predicted amounts of Fe precipitates were in good agreement with measured values for particulate Fe. Surface area and reactive site characteristics were used along with surface complexation constants for ferrihydrite (Dzombak, D. A.; Morel, F. M. M. Surface Complexation Modeling: Hydrous Ferric Oxide; John Wiley & Sons: New York, 1990) to predict ion sorption as a function of mixing fraction. Comparisons of model predictions with field results indicate that Pb and Cu sorption are predicted well by the model, while As, Mo, and Sb sorption are less well-predicted. Additional comparisons with particulate metal and Fe data collected from the CdA Mining District in 1996 and 1997 suggest that sorption on particulate Fe, including amorphous iron oxides and schwertmannite, may be described using universal model parameters. PMID- 11871566 TI - Arsenic(III) oxidation by birnessite and precipitation of manganese(II) arsenate. AB - Solution chemical techniques were used to investigate the oxidation of As(III) to As(V) in 0.011 M arsenite suspension of well-crystallized hexagonal birnessite (H birnessite, 2.7 g L(-1)) at pH 5. Products of the reaction were studied by scanning electron microscopy coupled with energy dispersive spectroscopy (SEM EDS), atomic force microscopy (AFM), and X-ray absorption near-edge structure spectroscopy (XANES). In the initial stage (first 74 h), chemical results have been interpreted quantitatively, and the reaction is shown to proceed in two steps as suggested by previous authors: 2>Mn(IV)O2 + H3AsO3 + H2O --> 2>Mn(III)OOH + H2AsO4- + H+ and 2>Mn(III)OOH + H3AsO3 + 3H+ --> 2Mn2+ + H2AsO4- + 2H2O. The As(III) depletion rate was lower (0.02 h(-1)) than measured in previous studies because of the high crystallinity of the H-birnessite sample used in this study. The surface reaction sites are likely located on the edges of H-birnessite layers rather than on the basal planes. The ion activity product of Mn(II) and As(V) reached after 74 h reaction time was the solubility product of a protonated manganese arsenate, having a chemical composition close to that of krautite as identified by XANES and EDS. Krautite precipitation reaction can be written as follows: Mn2+ + H2AsO4- + H2O = MnHAsO4 x H2O + H+ log Ks approximately -0.2. Equilibrium was reached after 400 h. The manganese arsenate precipitate formed long fibers that aggregated at the surface of H-birnessite. The oxidation reaction transforms a toxic species, As(III), to a less toxic aqueous species, which further precipitates with Mn2+ as a mixed As-Mn solid characterized by a low solubility product. PMID- 11871568 TI - Rapidly rising PBDE levels in North America. PMID- 11871567 TI - Sorption-desorption of ionogenic compounds at the mineral-water interface: study of metal oxide-rich soils and pure-phase minerals. AB - Sorption of the ionic compounds 2,4-D and quinmerac onto iron oxide-rich, variable charged soils was strongly influenced by mineralogy, particularly soil iron and aluminum oxides, whereas sorption of the neutral norflurazon was only related to total soil C. An appreciable fraction of the mass sorbed in stirred flow studies was easily desorbed by deionized water, and desorption of ionic compounds was initially more rapid than sorption. This sorption-desorption behavior, although contrary to desorption hysteresis commonly observed in batch studies, suggests that the reversibly sorbed fraction is weakly bound to the soil surface. 2,4-D sorption to iron oxide-rich soils and pure-phase metal oxides appears to be driven by nonspecific electrostatic attraction, with specific electrostatic attraction and van der Waals interactions being secondary. Both the carboxylate and the heterocyclic N groups may participate in sorption of quinmerac, facilitated by specific and nonspecific electrostatic attraction and surface complexation. The heterocyclic N, amine, and carbonyl groups of norflurazon do not appear to interact with soil minerals. PMID- 11871569 TI - Iron oxide surface-catalyzed oxidation of ferrous iron by monochloramine: implications of oxide type and carbonate on reactivity. AB - The maintenance of monochloramine residuals in drinking water distribution systems is one technique often used to minimize microbial outbreaks and thereby maintain the safety of the water. Reactions between oxidizable species and monochloramine can however lead to undesirable losses in the disinfectant residual. Previous work has illustrated that the Fe(II) present within distribution systems is one type of oxidizable species that can exert a monochloramine demand. This paper extends this prior work by examining the kinetics of the reactions between Fe(II) and monochloramine in the presence of a variety of iron oxide surfaces. The identity of the iron oxide plays a significant role in the rate of these reactions. Surface area-normalized initial rate coefficients (k(init)) obtained in the presence of each oxide at pH approximately 6.9 exhibit the following trend in catalytic activity: magnetite > goethite > hematite approximately = lepidocrocite > ferrihydrite. The differences in the activity of these oxides are hypothesized to result from variations in the amount of Fe(II) sorbed to each of the oxides and to dissimilarities in the surface site densities of the oxides. The implications of carbonate on Fe(II) sorption to iron oxides are also examined. Comparing Fe(II) sorption isotherms for goethite obtained under differential carbonate concentrations, it is apparent that as the carbonate concentration (C(T,CO3)) increased from 0 to 11.7 mM that the Fe(II) sorption edge (50% sorption) shifts from a pH of approximately 5.8 to a pH of 7.8. This shift is hypothesized to be the result of the formation of aqueous and surface carbonate-Fe(II) complexes and to competition between carbonate and Fe(II) for surface sites. The implications of these changes are then discussed in light of the variable oxide studies. PMID- 11871570 TI - U.S. electronics recycling stance challenged. PMID- 11871571 TI - Nutrient trading advocated to improve water quality. PMID- 11871573 TI - Congress boosts EPA's R&D funding. PMID- 11871572 TI - Atrazine linked to endocrine disruption in frogs. PMID- 11871574 TI - Dust storm provides clues to aerosol mixing. PMID- 11871575 TI - Profitably decontaminating sediments. PMID- 11871576 TI - Jim Morgan, an environmental visionary. PMID- 11871577 TI - Life-cycle assessment and the precautionary principle. PMID- 11871579 TI - The theory and measurement of the self-efficacy construct. AB - Self-efficacy is the major concept of Bandura's social cognitive theory. Self efficacy is influenced by four important sources of information: performance accomplishments, vicarious experience, verbal persuasion, and physiological information. Other determinants of self-efficacy are internal personal factors and external environmental factors. The degree of change in self-efficacy is partly a function of the variability and the controllability of its determinants. Level of self-efficacy predicts how people are functioning in terms of choice of behavior, effort expenditure and persistence, thought patterns and emotional reactions. Measurement of self-efficacy is related to three dimensions: magnitude, strength and generality. Self-efficacy should be measured in terms of particularized judgments of capability that may vary across realms of activity, different levels of task demands within a given activity domain, and different situational circumstances. PMID- 11871578 TI - Self-efficacy: measurement and intervention in nursing. AB - Assisting people with chronic illnesses to change their behavior is important in effecting self-management and in achieving the highest possible level of health. There is increasing evidence that a vital ingredient in health-related behavior change is the perceived self-efficacy of the individual to behave differently; however, disease- and age-specific measures and interventions have received insufficient attention to date. This special issue reports the results of the first stages of an international collaboration that is addressing the development and testing of instruments and interventions to measure and ultimately to enhance self-efficacy in management of diabetes mellitus. These instruments are now being used and tested further by other investigators, as well as in ongoing studies by the authors. Additionally, the issue addresses the importance of theory-based interventions and their use in enhancing self-efficacy as a means of encouraging positive behavior change. The results of two intervention studies designed to improve patients' self-management of obesity and cancer support the use of strategies to enhance self-efficacy in changing behavior. They also underscore the need to continue to identify, develop and test targeted nursing interventions. PMID- 11871580 TI - Self-efficacy in children with diabetes mellitus: testing of a measurement instrument. AB - The study reported here assessed the psychometric properties of an instrument to measure diabetes management self-efficacy in Dutch children, ages 8 to 12. Content validity of the item list was tested by consulting an expert panel of diabetes nurse specialists and a child-oriented rating scale was developed. A pretest was conducted to assess whether the instrument was clear and comprehensible for children. Then the instrument was tested in children with diabetes at the outpatient diabetes clinics for children at three Dutch hospitals. Reliability and criterion-related validity of the instrument were assessed. The instrument was judged content valid by the expert panel of diabetes nurse specialists, and the language of the instructions, the items and the rating scale was found to be clear and understandable for children of this age group. The study yielded only a moderate internal consistency estimate (Cronbach's = 0.71) and limited support for criterion-related validity. Several useful theoretical and methodological issues were identified. PMID- 11871581 TI - The development and psychometric testing of an instrument to measure diabetes management self-efficacy in adolescents with type 1 diabetes. AB - Adherence to treatment is a major issue for people with diabetes mellitus, and attention has been given to improving self-management in persons with this chronic disease. Most studies show that knowledge alone will not improve self management behavior. Self-efficacy, a concept introduced by Bandura, has been shown to be an important variable in improving health behaviors. In order to determine whether self-efficacy can influence self-management of diabetes, however, instruments to measure self-efficacy are needed in native languages at an appropriate developmental level, and with good psychometric properties. The study reported here was part of a larger project in the Netherlands, Belgium and the United States in which several instruments, measuring diabetes management self-efficacy, have been developed, both in English and Dutch, for different types of diabetes and different age groups. This article reports on the development and psychometric testing of an instrument measuring self-efficacy in adolescents with type 1 diabetes mellitus. Initially 30 items for the instrument were generated through focus group interviews and their relevance was judged by a team of 10 experts on self-management behavior in adolescents. The final instrument contained 26 items. The sample for psychometric testing consisted of 90 patients with type 1 diabetes who were between 12 and 18 years old. Cronbach's alpha of the instrument was 0.86. An exploratory factor analysis produced two factors which reflect general and more difficult diabetes self-management situations. PMID- 11871582 TI - Strategies enhancing self-efficacy in diabetes education: a review. AB - Enhancing self-efficacy in patients with chronic illnesses has been shown to have a positive effect on behavior change. In fact, according to Bandura (1986), self efficacy is the most important predictor of change in behavior. Thus, in order to make positive changes, effective measures for enhancing self-efficacy are needed in educational programs. There are four important sources of information for increasing self-efficacy: performance accomplishments, vicarious learning, verbal persuasion, and self-appraisal of emotional and physiological responses. Strategies for enhancing self-efficacy are described here for each source of information and for combinations of sources. PMID- 11871583 TI - The use of self-efficacy enhancing methods in diabetes education in the Netherlands. AB - According to the social cognitive theory of Bandura, self-efficacy predicts behavorial change. Bandura notes that self-efficacy is based on four major sources of information: performance accomplishments, vicarious experience, verbal persuasion and self-evaluation. This exploratory study examined the use of these four sources of information by Dutch nurse diabetes educators to enhance self efficacy among people with diabetes mellitus. A survey questionnaire was sent to all Dutch nurse members of the European Association of Diabetes Educators (EADE) asking about the use of self-efficacy-enhancing methods, and four different educational programs were observed. Survey respondents said that performance accomplishments and verbal persuasion were often used, vicarious experience was hardly ever used, and the use of self-evaluation varied. The observations gave a different picture: only verbal persuasion was observed often; the other three sources were hardly ever used. Clearly, self-efficacy-enhancing educational methods are not systematically used in the Netherlands and there is little variety in the methods used. More varied methods of enhancing self-efficacy need to be developed and implemented in diabetes education programs. PMID- 11871584 TI - Self-efficacy targeted treatments for weight loss in postmenopausal women. AB - Matching behavioral treatment programs to different types of obese clients is a rarely studied strategy, thus guidelines for identifying who might benefit most from which program remain elusive. This study categorized the weight control self efficacy beliefs of obese, postmenopausal women, and determined the effects of self-efficacy targeted versus non-targeted (control) treatment on weight loss outcomes (body habitus, physical conditioning, affect, and eating behaviors). Obese (BMI 33 +/- 5), postmenopausal (60 +/- 6 years old) women (n = 59) participated in a 6-month weight loss program. The 37 women categorized by Q methodology as Assured (self-confident, independent) were randomized to Assured (AT) or Non-Targeted (NT) treatment; the 22 Disbelievers (doubtful, wavering) were randomized to Disbeliever (DT) or NT treatment. At baseline, the Assureds had significantly smaller body girths and reported significantly greater self esteem, fewer symptoms of binge eating, and less negative affect overeating than the Disbelievers. Improvement in these variables with weight loss erased significant differences between the groups and was a desired outcome. Treatment delivery type may have influenced attrition rate, since significantly more Assureds dropped from NT than AT and significantly more Disbelievers dropped from DT than NT. Thus, the self-efficacy type may serve as a means to identify different types of treatment needs (flexible vs. rigid) to sustain women's adherence and success in the program. The significant weight loss outcomes for women in all groups argues for the incorporation of strategies to enhance self efficacy but not the need for specific treatments that directly target self efficacy types. PMID- 11871585 TI - An intervention to increase quality of life and self-care self-efficacy and decrease symptoms in breast cancer patients. AB - This study tested effects of a nurse-administered self-efficacy intervention given on five monthly occasions and designed to enhance patients' self-care self efficacy. The hypotheses were that at four months and eight months after beginning chemotherapy the efficacy-enhancing experimental group would have significantly higher scores on quality of life and self-care self-efficacy than the control group and significantly less symptom distress. Fifty-six women receiving chemotherapy for breast cancer were randomized to the experimental and control groups. Outcome variables were quality of life, measured by the Functional Assessment of Cancer Treatment-Breast (FACT-B), symptom distress, measured by the Symptom Distress Scale (SDS), and factors of self-care self efficacy, measured by Strategies Used by Patients to Promote Health (SUPPH). The interaction effects for the FACT-B ranged from small for functional concerns (eta square = .03) to large for social concerns (eta square = .110); effects for the SDS were large (eta square = .140), and for factors on the SUPPH effect sizes ranged from small (eta square = .01) for Enjoying Life and Stress Reduction to medium (eta square = .089) for Coping, and large (eta square = .141) for Making Decisions. Interventions to promote self-efficacy may increase quality of life and decrease symptom distress for women diagnosed with breast cancer. PMID- 11871587 TI - Problems with the performance of the SF-36 among people with type 2 diabetes in general practice. AB - OBJECTIVE: To validate the short form-36 (SF-36) among people with type 2 diabetes in general practice, and to make comparisons with the Audit of Diabetes Dependent Quality of Life (ADDQoL). DESIGN: Postal survey with one reminder. SETTING: Four general practices. PATIENTS: One hundred and eighty-four eligible patients (30-70 years) with type 2 diabetes on 14 general practitioner lists. MEASURES: SF-36 response rates, distribution of dimension scores and internal consistency. Median scores in relation to sociodemography and self-reported health. Comparisons with ADDQoL scores. RESULTS: One hundred and thirty-one patients responded (71%). Distributions of SF-36 dimension scores were mostly skewed. Internal consistency and construct validity were acceptable, with predictable sociodemographic trends. People with illness related to or unrelated to diabetes scored significantly lower on most dimensions. SF-36 dimension scores correlated best with relevant diabetes-specific ADDQoL scores amongst respondents reporting no comorbidity. CONCLUSIONS: Although valid and reliable, SF-36 scores are strongly affected by non-diabetic comorbidity in type 2 diabetes, supporting the complementary use of a diabetes-specific measure, providing information about the impact of diabetes specifically. PMID- 11871586 TI - Modeling changes in health perception following hip fracture. AB - BACKGROUND AND OBJECTIVE: Health perception is an important outcome associated with health-related quality of life. Various correlates of perceived health have been identified, but attempts at multivariate modeling have often failed to mirror the complex system of causal pathways surrounding this concept. The objective of this study was to develop a multivariate model to improve our understanding of how physical, social, and psychological factors interact to influence health perception following a hip fracture. METHODS: Patients were interviewed in-hospital during the week following the fracture to obtain information on in-hospital mental status and pre-fracture function, pre-fracture health perception, and pre-fracture social support. A follow-up interview was conducted by telephone to assess function, health perception, and social support 3 months post-fracture. Comorbidities and demographic information were obtained from medical records. RESULTS: This study included 222 hip fracture patients aged 65 + and fracturing a hip between 10 July 1996 and 30 August 1997. Our estimated model was theory-based, developed from existing research identifying the following correlates of health perception: physical function, comorbidities, socioeconomic status, social support, age, and prior health perception. In addition to these correlates, our model was unique in incorporating a variable to assess whether the gap between pre- and post-fracture functioning acted as a separate cause on post-fracture health perception. Initially, the fit between the data and the model was poor, however minor modifications to the model corrected this. While there was a good fit between the data and the estimated model, only about 25% of the variation in both pre- and post-fracture health perceptions were explained. The number of comorbidities and physical function were found to have the strongest influence on health perception. Post-fracture health perceptions were also improved by increased post-fracture social support, higher income/status, and higher pre-fracture health perception. The network of effects indicates that the causal sources of health perception may differ in the pre- and post-fracture periods. This suggests that the causal sources of health perception following major health changes may differ substantially from the causal sources of health perception operating during normal times. CONCLUSIONS: The findings suggest that interventions aimed at improving pre-fracture function and post fracture social support could increase health perception following hip fracture. In interpreting our results, it is important to remember that while we attempted to adjust for measurement error in the analysis, the study may be limited due to the retrospective nature of the questions. Future research should focus on improving the model by including other components of the patient's emotional health. PMID- 11871588 TI - Social support and health-related quality of life in chronic heart failure patients. AB - OBJECTIVES: Objectives of this study were to: (1) describe perceived social support during a baseline hospitalization and 12 months later among heart failure patients; (2) examine differences in social support as a function of gender and age (less than 65 and 65 years or older); and (3) examine social support as a predictor of health-related quality of life. BACKGROUND: Social support is a predictor of well-being and mortality, but little is known about support patterns among heart failure patients and how they influence quality of life. METHODS: The sample included 227 hospitalized patients with heart failure who completed the Social Support Survey and the Chronic Heart Failure Questionnaire at baseline; 147 patients completed these questionnaires again 12 months after baseline. RESULTS: Mean baseline and 12-month total support scores were 56 and 53, respectively, with a score of 76 indicating the most positive perceptions of support. The ANOVA indicated significant interactions of gender by age for total (F = 5.04; p = 0.03) and emotional/informational support (F = 4.87; p = 0.03) and for positive social interactions (F = 4.43; p = 0.04), with men under age 65 perceiving less support than men aged 65 and older and women in either age group. Baseline support did not predict 12-month health-related quality of life, but changes in social support significantly predicted changes in health-related quality of life (R2 = 0.14). CONCLUSIONS: Overall, perceptions of support were moderate to high, but there was wide variation in perceptions over time. Men under age 65 reported less support than other groups of patients. Importantly, changes in social support were significant predictors of changes in health related quality of life. PMID- 11871589 TI - Does electronic implementation of questionnaires used in asthma alter responses compared to paper implementation? AB - BACKGROUND: Electronic implementation of questionnaires has many advantages, but there may be concerns that it alters versions that were validated on paper. OBJECTIVE: To determine whether electronic implementation alters responses to the SF-36 and asthma quality of life questionnaire (AQLQ), compared to paper implementation. METHODS: Patients with asthma presenting to a pneumologist were asked for consent to participate. Each patient completed both forms of each questionnaire. The order of presentation was alternated sequentially, with the first patient completing the electronic version first. Each patient waited at least 2 hours between completions to minimize recollection of answers. For both the SF-36 and AQLQ, intraclass correlations coefficients were calculated to compare patients' scores, for each scale and overall, on the electronic and paper versions. RESULTS: Sixty-eight patients (mean age: 48 years, 50 females) of 311 contacted were enrolled. Overall intraclass correlation coefficients for the SF 36 and AQLQ were excellent (0.965 and 0.991 respectively). For paper versions, eight questions (AQLQ) and 24 (SF-36) were left blank and nine questions (SF-36) were answered incorrectly by patients selecting more than one answer. Electronic data for one patient could not be retrieved. CONCLUSION: Collecting SF-36 and AQLQ data electronically can decrease the number of spoiled responses without altering the results. Successful implementation depends on proper instruction of the respondent in the handling of the electronic instrument. PMID- 11871590 TI - Health related quality of life in patients with venous ulceration: use of the Nottingham health profile. AB - INTRODUCTION: The Nottingham health profile (NHP) has been used in several studies of patients being treated for chronic venous leg ulceration, though there is a paucity of information on the validity of the NHP in this patient group. This study was carried out to determine this validity and to compare and contrast the results with a previous study that had used the MOS short form 36 (SF-36) in this patient group. METHODS: Patients in four health trusts suffering from venous ulceration were interviewed at entry and after 12 weeks of treatment using high compression bandaging. Information was collected on ulcer history, general medical history, and the NHP was completed at both time points. After 12 weeks, a record was made of whether all areas of ulceration had cleared from the patients' legs (healed) or whether some ulceration remained (unhealed). RESULTS: Of the 383 patients who made up the study population, 37% experienced complete healing of their ulceration after 12 weeks of treatment. While internal consistencies of the NHP scores were reasonable (Cronbach's alpha > 0.63), there were a high proportion of patients who scored 0 (best possible health) at entry into the study, most notably for social isolation (67.5%), emotional status (48.9%) and energy (47.4%). Despite these limitations, there was strong evidence that treatment led to significant improvements in all dimensions of the NHP for the total group (all p < 0.01). These improvements were greatest in patients whose ulcers healed compared with patients whose ulcers failed to heal for bodily pain (mean difference d = 9.4, p = 0.004). Patients whose overall health status improved had significantly greater improvements for all dimensions of the NHP (p < 0.05). CONCLUSION: The NHP, has limitations in terms of a large floor effect (best possible health), but has similar internal consistency to the SF-36 in patients with leg ulceration. However, unlike the SF-36, the NHP is sensitive to change in the patients' ulcer status, and should therefore be considered in studies of venous leg ulcer healing. PMID- 11871591 TI - Cross-cultural validation of an international questionnaire, the General Measure of the Functional Assessment of Cancer Therapy scale (FACT-G), for Japanese. AB - The General Measure of the Functional Assessment of Cancer Therapy scale (FACT-G) was developed in an English-speaking culture (USA). To determine if FACT-G could be used in Japan, a cross-cultural validation was performed. The Japanese version was created through an iterative forward-backward translation sequence used throughout the FACT multi-lingual translation project. In evaluating psychometric testing, its construct validity was investigated by factor analysis and multi trait scaling analysis. Clinical validity was estimated by known-groups comparison using stage, performance score (PS) and patient location, and validated longitudinally by PS. The FACT-G (version 3) was given to 180 patients with lung cancer. Analyses showed that the scales of Physical, Functional, Emotional Well-Being, and Relationship with Doctors were constructively valid in Japan. Japanese patients felt that familial relationships were different than those with friends and neighbors, indicating that the Social/Family Well-Being scale needed cultural adaptation. Two items concerning coping with illness and acceptance of illness did not load predictably onto their respective scales and were considered to be cross-culturally problematic. However, clinical validity demonstrated its sensitivity. Japanese version 4 has been improved to address the weakness in an attempt to become an instrument that is applicable across cultures. PMID- 11871592 TI - The WHO quality of life assessment instrument (WHOQOL-Bref): the importance of its items for cross-cultural research. AB - One of the fundamental issues in the area of assessment of quality of life is to determine what is important to the individuals' quality of life. This is even more crucial when the instrument is for use in diverse cultural settings. This paper reports on the importance ratings on WHOQOL-Bref items obtained as a part of WHOQOL pilot field trial on 4804 respondents from 15 centres from 14 developed and developing countries using 12 languages. All items were rated as moderately or more important, but this was expected because the items were selected by extensive qualitative research for their salience across the centres. Significant differences on mean importance ratings were found between centres, but rank orders of item for their importance showed highly significant correlations between centres. This was especially true for items in the top and the bottom thirds of the item list arranged by overall importance. Most items were rated as more important by women compared to men and by younger compared to older persons. The results are discussed for their relevance in cross-cultural research on quality of life assessment. PMID- 11871593 TI - Internal consistency, reproducibility, responsiveness, and construct validity of the Chinese (HK) version of the asthma quality of life questionnaire. AB - The purpose of this study was to test the acceptability, psychometric properties and construct validity of a Chinese translation of the Asthma Quality of Life Questionnaire (AQOL) and to determine whether it has any advantage over the Chinese (HK) version of the generic MOS SF-36 Health Survey that has been validated on Chinese in Hong Kong. Fifty-two patients with asthma were interviewed using the Chinese (HK) AQOL and the Chinese (HK) SF-36. Thirty-four of the 52 patients were interviewed a second time 6 months later using the same questionnaires. The Cronbach's alpha coefficients were above 0.7 for all four domains of the AQOL questionnaire. Significant correlations were found between the scores of each of the four domains of the Chinese (HK) AQOL and forced expiratory volume in 1 s (FEV1) (%predicted) during the initial visit and between changes in the mean scores of most of the domains of Chinese (HK) AQOL and changes in FEV1 between visits. The Chinese (HK) AQOL was found to be reproducible in those with stable asthma, and for those whose asthma severity changed between visits, there was significant change in one of the domains. It can detect differences in quality of life in asthma patients of varying severity. We conclude that the Chinese (HK) translation of the AQOL is reliable, valid, reproducible and discriminative and can be used to supplement generic instruments in evaluating the quality of life of patients with asthma. PMID- 11871594 TI - Computational analysis of coupled blood-wall arterial LDL transport. AB - The transport of macromolecules, such as low density lipoproteins (LDLs), across the artery wall and their accumulation in the wall is a key step in atherogenesis. Our objective was to model fluid flow within both the lumen and wall of a constricted, axisymmetric tube simulating a stenosed artery, and to then use this flow pattern to study LDL mass transport from the blood to the artery wall. Coupled analysis of lumenal blood flow and transmural fluid flow was achieved through the solution of Brinkman's model, which is an extension of the Navier-Stokes equations for porous media. This coupled approach offers advantages over traditional analyses of this problem, which have used possibly unrealistic boundary conditions at the blood-wall interface; instead, we prescribe a more natural pressure boundary condition at the adventitial vasa vasorum, and allow variations in wall permeability due to the occurrence of plaque. Numerical complications due to the convection dominated mass transport process (low LDL diffusivity) are handled by the streamline upwind/Petrov-Galerkin (SUPG) finite element method. This new fluid-plus-porous-wall method was implemented for conditions typical of LDL transport in a stenosed artery with a 75 percent area reduction (Peclet number=2 x 10(8)). The results show an elevated LDL concentration at the downstream side of the stenosis. For the higher Darcian wall permeability thought to occur in regions containing atheromatous lesions, this leads to an increased transendothelial LDL flux downstream of the stenosis. Increased transmural filtration in such regions, when coupled with a concentration-dependent endothelial permeability to LDL, could be an important contributor to LDL infiltration into the arterial wall. Experimental work is needed to confirm these results. PMID- 11871595 TI - Experimental optimization of pivot point height for swing-arm type rear suspensions in off-road bicycles. AB - Towards the ultimate goal of designing dual suspension off-road bicycles which decouple the suspension motion from the pedaling action, this study focused on determining experimentally the optimum pivot point height for a swing-arm type rear suspension such that the suspension motion was minimized. Specific objectives were (1) to determine the effect of interaction between the front and rear suspensions on the optimal pivot point height, (2) to investigate the sensitivity of the optimal height to the pedaling mechanics of the rider in both the seated and standing postures, (3) to determine the dependence of the optimal height on the rider posture. Eleven experienced subjects rode a custom-built adjustable dual suspension off-road bicycle, [Needle, S., and Hull, M. L., 1997, "An Off-Road Bicycle With Adjustable Suspension Kinematics," Journal of Mechanical Design 119, pp. 370-375], on an inclined treadmill. The treadmill was set to a constant 6 percent grade at a constant velocity of 24.8 km/hr. With the bicycle in a fixed gear combination of 38 x 14, the corresponding cadence was 84 rpm. For each subject, the pivot point height was varied randomly while the motions across both the front and rear suspension elements were measured. Subjects rode in both the seated and standing postures and with the front suspension active and inactive. It was found that the power loss from the rear suspension at the optimal pivot point height was not significantly dependent on the interaction between the front and rear suspensions. In the seated posture, the optimal pivot point height was 9.8 cm on average and had a range of 8.0-12.3 cm. The average optimal pivot point height for the seated posture corresponded to an average power loss for the rear suspension that was within 10 percent of the minimum power loss for each subject for 8 of the 11 subjects. In the standing posture, the average height was 5.9 cm and ranged from 5.1-7.2 cm. The average heightfor the standing posture was within 10 percent of the minimum power loss for each subject for 9 of the 11 subjects. While the optimum height was relatively insensitive to pedaling mechanics in both the seated and standing postures, the choice of the optimal pivot point height in production bicycles necessitates some compromise in performance given the disparity in the averages between the seated and standing postures. PMID- 11871596 TI - Biomechanical factors affecting the peak hand reaction force during the bimanual arrest of a moving mass. AB - Fall-related wrist fractures are among the most common fractures at any age. In order to learn more about the biomechanical factors influencing the impact response of the upper extremities, we studied peak hand reaction force during the bimanual arrest of a 3.4 kg ballistic pendulum moving toward the subject in the sagittal plane at shoulder height. Twenty healthy young and 20 older adults, with equal gender representation, arrested the pendulum after impact at one of three initial speeds: 1.8, 2.3, or 3.0 m/sec. Subjects were asked to employ one of three initial elbow angles: 130, 150, or 170 deg. An analysis of variance showed that hand impact force decreased significantly as impact velocity decreased (50 percent/m/s) and as elbow angle decreased (0.9 percent/degree). A two segment sagittally-symmetric biomechanical model demonstrated that two additional factors affected impact forces: hand-impactor surface stiffness and damping properties, and arm segment mass. We conclude that hand impact force can be reduced by more than 40 percent by decreasing the amount of initial elbow extension and by decreasing the velocity of the hands and arms relative to the impacting surface. PMID- 11871598 TI - A new fundamental bioheat equation for muscle tissue--part II: Temperature of SAV vessels. AB - In this study, a new theoretical framework was developed to investigate temperature variations along countercurrent SAV blood vessels from 300 to 1000 microm diameter in skeletal muscle. Vessels of this size lie outside the range of validity of the Weinbaum-Jiji bioheat equation and, heretofore, have been treated using discrete numerical methods. A new tissue cylinder surrounding these vessel pairs is defined based on vascular anatomy, Murray's law, and the assumption of uniform perfusion. The thermal interaction between the blood vessel pair and surrounding tissue is investigated for two vascular branching patterns, pure branching and pure perfusion. It is shown that temperature variations along these large vessel pairs strongly depend on the branching pattern and the local blood perfusion rate. The arterial supply temperature in different vessel generations was evaluated to estimate the arterial inlet temperature in the modified perfusion source term for the s vessels in Part I of this study. In addition, results from the current research enable one to explore the relative contribution of the SAV vessels and the s vessels to the overall thermal equilibration between blood and tissue. PMID- 11871597 TI - Energetics of actively powered locomotion using the simplest walking model. AB - We modified an irreducibly simple model of passive dynamic walking to walk on level ground, and used it to study the energetics of walking and the preferred relationship between speed and step length in humans. Powered walking was explored using an impulse applied at toe-off immediately before heel strike, and a torque applied on the stance leg. Although both methods can supply energy through mechanical work on the center of mass, the toe-off impulse is four times less costly because it decreases the collision loss at heel strike. We also studied the use of a hip torque on the swing leg that tunes its frequency but adds no propulsive energy to gait. This spring-like actuation can further reduce the collision loss at heel strike, improving walking energetics. An idealized model yields a set of simple power laws relating the toe-off impulses and effective spring constant to the speed and step length of the corresponding gait. Simulations incorporating nonlinear equations of motion and more realistic inertial parameters show that these power laws apply to more complex models as well. PMID- 11871599 TI - The modulus of toughness of urinary calculi. AB - To improve the efficacy of extracorporeal shock wave lithotripsy (ESWL) treatment, it is desirable to identify the physical properties of urinary calculi could offer direct correlation with their fragilities during ESWL and thus could be used to guide treatment procedures for more effective stone fragmentation. Thirty stone specimens removed surgically were compressed by an axial testing system to measure the compressive strength and trace the stress-strain curve. Image analysis software SigmaScan (Jandel Co.) was used to calculate the area under the stress-strain curve, the modulus of toughness, for each stone. The values of compressive strength measured were similar to those reported by other researchers. The modulus of toughness of urinary calculi correlates with clinical representation of the stone fragility during ESWL. The modulus of toughness could be an index to evaluate the physical property of urinary calculi that could be used to guide treatment procedures for more effective stone fragmentation. PMID- 11871600 TI - Fitting manifold surfaces to three-dimensional point clouds. AB - We present a technique for fitting a smooth, locally parameterized surface model (called the manifold surface model) to unevenly scattered data describing an anatomical structure. These data are acquired from medical imaging modalities such as CT scans or MRI. The manifold surface is useful for problems which require analyzable or parametric surfaces fitted to data acquired from surfaces of arbitrary topology (e.g., entire bones). This surface modeling work is part of a larger project to model and analyze skeletal joints, in particular the complex of small bones within the wrist and hand. To demonstrate the suitability of this model we fit to several different bones in the hand, and to the same bone from multiple people. PMID- 11871601 TI - Unsteady effects on the flow across tilting disk valves. AB - The present study simulates numerically the flow across two-dimensional tilting disk models of mechanical heart valves. The time-dependent Navier-Stokes equations are solved to assess the importance of unsteady effects in the fully open position of the valve. Flow cases with steady or physiological inflow conditions and with fixed or moving valves are solved. The simulations lead into mixed conclusions. It is obvious that steady inflow cases that account for vortex shedding only cannot model realistic physiological cases. In cases with imposed physiological inflow, the details of the flow field for fixed and moving valves might differ in the fully open position as well, although the gross features are quite similar. The fixed valve case consistently results in safe estimations of several critical quantities such as the axial force, the maximal shear stress on the valve, or the transvalvular pressure drop. Thus, fixed valve simulations can provide useful information for the design of prosthetic heart valves, as long as the properties in the fully open position only are sought. PMID- 11871602 TI - Control of the shape of a thrombus-neointima-like structure by blood shear stress. AB - Fluid mechanical factors are thought to influence vascular morphogenesis. Here we show how blood shear stress regulates the shape of a thrombus-neointima-like tissue on a polymer micro-cylinder implanted in the center of the rat vena cava with the micro-cylinder perpendicular to blood flow. In this model, the micro cylinder is exposed to a laminarflow with a known shear stress field in the leading region and a vortexflow in the trailing region. At 1, 5, 10, 20, and 30 days after implantation, it was found that the micro-cylinder was encapsulated by a thrombus-neointima-like tissue with a streamlined body profile. The highest growth rate of the thrombus-neointima-like tissue was found along the trailing and leading stagnation edges of the micro-cylinder. Blood shear stress in the laminar flow region was inversely correlated with the rate of thrombus formation and cell proliferation, and the percentage of smooth muscle a actin-positive cells. These biological changes were also found in the trailing vortex flow region, which was associated with lowered shear stress. These results suggest that blood shear stress regulates the rate of thrombus and neointimal formation and, thus, influences the shape of the thrombus-neointima-like structure in the present model. PMID- 11871603 TI - Fluid shear stress-induced alignment of cultured vascular smooth muscle cells. AB - The study objectives were to quantify the time- and magnitude-dependence of flow induced alignment in vascular smooth muscle cells (SMC) and to identify pathways related to the orientation process. Using an intensity gradient method, we demonstrated that SMC aligned in the direction perpendicular to applied shear stress, which contrasts with parallel alignment of endothelial cells under flow SMC alignment varied with the magnitude of and exposure time to shear stress and is a continuous process that is dependent on calcium and cycloskeleton based mechanisms. A clear understanding and control of flow-induced SMC alignment will have implications for vascular tissue engineering. PMID- 11871604 TI - Relative contribution of wall shear stress and injury in experimental intimal thickening at PTFE end-to-side arterial anastomoses. AB - BACKGROUND: Intimal hyperplastic thickening (IHT) is a frequent cause of prosthetic bypass graft failure. Induction and progression of IHT is thought to involve a number of mechanisms related to variation in the flow field, injury and the prosthetic nature of the conduit. This study was designed to examine the relative contribution of wall shear stress and injury to the induction of IHT at defined regions of experimental end-to-side prosthetic anastomoses. METHODS AND RESULTS: The distribution of IHT was determined at the distal end-to-side anastomosis of seven canine Iliofemoral PTFE grafts after 12 weeks of implantation. An upscaled transparent model was constructed using the in vivo anastomotic geometry, and wall shear stress was determined at 24 axial locations from laser Doppler anemometry measurements of the near wall velocity under conditions of pulsatile flow similar to that present in vivo. The distribution of IHT at the end-to-side PTFE graft was determined using computer assisted morphometry. IHT involving the native artery ranged from 0.0+/-0.1 mm to 0.05+/ 0.03 mm. A greater amount of IHT was found on the graft hood (PTFE) and ranged from 0.09+/-0.06 to 0.24+/-0.06 mm. Nonlinear multivariable logistic analysis was used to model IHT as a function of the reciprocal of wall shear stress, distance from the suture line, and vascular conduit type (i.e. PTFE versus host artery). Vascular conduit type and distance from the suture line independently contributed to IHT. An inverse correlation between wall shear stress and IHT was found only for those regions located on the juxta-anastomotic PTFE graft. CONCLUSIONS: The data are consistent with a model of intimal thickening in which the intimal hyperplastic pannus migrating from the suture line was enhanced by reduced levels of wall shear stress at the PTFE graft/host artery interface. Such hemodynamic modulation of injury induced IHT was absent at the neighboring artery wall. PMID- 11871606 TI - Effect of vessel compliance on the in-vitro performance of a pulsating respiratory support catheter. AB - Intravena caval respiratory support (or membrane oxygenation) is a potential therapy for patients with acute respiratory insufficiency. A respiratory support catheter is being developed that consists of a bundle of hollow fiber membranes with a centrally positioned pulsating balloon to enhance gas exchange. This study examined the influence of vessel compliance on the gas exchange performance of the pulsating respirator, support catheter. Polyurethane elastic tubes were fabricated with compliance comparable to that measured in bovine vena cava specimens. The gas exchange performance of the respiratory catheter was studied in an in-vitro flow loop using either the model compliant tube or a rigid tube as a "mock" vena cava. Balloon pulsation enhanced gas exchange comparably in both rigid and model compliant vessels up to 120 bpm pulsation frequency. Above 120 bpm gas exchange increased with further pulsation in the rigid tube, but no additional increase in gas exchange was seen in the compliant tube. The differences above 120 bpm may reflect differences in the compliance of the elastic tube versus the natural vena cava. PMID- 11871607 TI - Recruitment of tendon crimp with applied tensile strain. AB - The tensile stress-strain behavior of ligaments and tendons begins with a toe region that is believed to result from the straightening of crimped collagen fibrils. The in situ mechanical function is mostly confined to this toe region and changes in crimp morphology are believed to be associated with pathological conditions. A relatively new imaging technique, optical coherence tomography (OCT), provides a comparatively inexpensive method for nondestructive investigation of tissue ultrastructure with resolution on the order of 15 microm and the potential for use in a clinical setting. The objectives of this work were to assess the utility of OCT for visualizing crimp period, and to use OCT to determine how crimp period changed as a function of applied tensile strain in rat tail tendon fascicles. Fascicles from rat tail tendons were subjected to 0.5 percent strain increments up to 5 percent and imaged at each increment using OCT. A comparison between OCT images and optical microscopy images taken between crossed polarizing lenses showed a visual correspondence between features indicative of crimp pattern. Crimp pattern always disappeared completely before 3 percent axial strain was reached. Average crimp period increased as strain increased, but both elongation and shortening occurred within single crimp periods during the application of increasing strain to the fascicle. PMID- 11871605 TI - A left ventricle model to predict post-revascularization ejection fraction based on cine magnetic resonance images. AB - The left ventricular ejection fraction (LVEF) is an important clinical indicator of the cardiac function and long-term outcome for patients with coronary artery disease. A biomechanical model of the left ventricle was developed to quantitatively predict post-revascularization LVEF based on noninvasive magnetic resonance imaging. The myocardium was categorized into normal, hibernating, and infarcted regions from the ventricular short-axis images. Assuming that hibernating tissue would potentially regain contractility after revascularization, the expected maximum post-revascularization LVEF was calculated for four patients with chronic left ventricular dysfunction. The predictions were within three ejection fraction points of the follow-up LVEFs. This model may be useful to estimate the outcome and efficacy of revascularization plans. PMID- 11871608 TI - Time-dependent mechanical behavior of sheep digital tendons, including the effects of preconditioning. AB - The time-dependent mechanical properties of sheep digital extensor tendons were studied by sequences of stress-relaxation tests. The results exhibited irreversible preconditioning and reversible viscoelasticity. Preconditioning effects were manifested by stress decay during consecutive stretch cycles to the same strain level, accompanied by elongation of the tendon's reference length. They intensified with increased strain level, and were reduced or became negligible as the strain decreased. The significance of intrinsic response mechanisms was studied via a structural model that includes viscoelasticity, preconditioning, and morphology of the tendon's collagen fibers. Model/data comparisons showed good agreement and good predictive power, suggesting that preconditioning can be integrated into comprehensive material characterization of tendons. PMID- 11871610 TI - Numerical analysis of flow through a severely stenotic carotid artery bifurcation. AB - The results of computational simulations may supplement MR and other in vivo diagnostic techniques to provide an accurate picture of the hemodynamics in particular vessels, which may help demonstrate the risks of embolism or plaque rupture posed by particular plaque deposits. In this study, a model based on an endarterectomy specimen of the plaque in a carotid bifurcation was examined. The flow conditions include steady flow at Reynolds numbers of 300, 600, and 900 as well as unsteady pulsatile flow. Both dynamic pressure and wall shear stress are very high, with shear values up to 70 N/m2, proximal to the stenosis throat in the internal carotid artery, and both vary significantly through the flow cycle. The wall shear stress gradient is also strong along the throat. Vortex shedding is observed downstream of the most severe occlusion. Two turbulence models, the Chien and Goldberg varieties of k-epsilon, are tested and evaluated for their relevance in this geometry. The Chien model better captures phenomena such as vortex shedding. The flow distal to stenosis is likely transitional, so a model that captures both laminar and turbulent behavior is needed. PMID- 11871609 TI - Effects of static stress on the mechanical properties of cultured collagen fascicles from the rabbit patellar tendon. AB - In-vitro tissue culture experiments were performed to study the effects of static stress on the mechanical properties of collagen fascicles obtained from the rabbit patellar tendon. After collagen fascicles having the diameter of approximately 300 microm were cultured for 1 and 2 wk under static stress between 0 and 3 MPa, their mechanical properties and crimp morphology were determined using a micro-tensile tester and a light microscope, respectively. The tensile strength and tangent modulus of the fascicles were significantly decreased by culture under no load compared to control fascicles. A statistically significant correlation, which was described by a quadratic curve, was observed between applied stress and tensile strength. The maximum tensile strength (16.7 MPa) was obtained at the applied stress of 1.2 MPa; the strength was within a range of control values. There was a similar correlation between applied stress and tangent modulus, and the modulus was maintained at control level under 1.3 MPa stress. The stress of 1.2 to 1.3 MPa is equivalent to approximately 50 percent of the peak stress developed in the intact rabbit patellar tendon by running. Strain at failure of cultured collagen fascicles was negatively correlated with applied stress, and that at 1.2 to 1.3 MPa stress was almost the same as the control value. Crimp morphology in the fascicles cultured under about 1.2 MPa stress was similar to that in control fascicles. These results indicate that cultured collagen fascicles change the mechanical properties and structure in response to static tensile stress. In addition, their mechanical properties and structure are maintained at control level if the static stress of 50 percent of in-vivo peak stress is applied. PMID- 11871611 TI - The mechanical advantage of local longitudinal shortening on peristaltic transport. AB - Whereas bolus transport along the esophagus results from peristaltic contractions of the circular muscle layer, it has been suggested that local shortening of the longitudinal muscle layer concentrates circular muscle fibers in the region where the highest contractile pressures are required. Here we analyze the mechanical consequences of local longitudinal shortening (LLS) through a mathematical model based on lubrication theory. We find that local pressure and shear stress in the contraction zone are greatly reduced by the existence of LLS. In consequence, peak contractile pressure is reduced by nearly 2/3 at physiological LLS, and this reduction is greatest when peak in LLS is well aligned with peak contractile pressure. We conclude that a peristaltic wave of local longitudinal muscle contraction coordinated with the circular muscle contraction wave has both a great physiological advantage (concentrating circular muscle fibers), and a great mechanical advantage (reducing the level of contractile force required to transport the bolus), which combine to greatly reduce circular muscle tone during esophageal peristalsis. PMID- 11871612 TI - Melanins in fungal pathogens. PMID- 11871613 TI - Rapid antimicrobial susceptibility testing of urinary tract isolates and samples by flow cytometry. AB - A multiparametric flow cytometry antimicrobial susceptibility test was developed and its performance was evaluated on clinical urine isolates and samples in comparison with standard methods. Alterations in cytoplasmic membrane integrity were monitored by propidium iodide, and the anionic probe bis-(1,3 dibutylbarbituric acid) trimethine oxonol (DiBAC4(3)) was used to measure changes in membrane potential. Microbial size and cellular content were analysed by light scattering. Twelve antibiotics were tested on 6 ATCC control strains, 22 urine isolates and 19 clinical urine samples, variously containing Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, Proteus mirabilis, Enterococcus faecalis, Staphylococcus aureus, S. saprophyticus and S. epidermidis. Agreement between the flow cytometry results, broth microdilution and disk diffusion tests was 93.9% (n = 328 tests). Of the 20 discrepancies observed, 18 were for species other than E. coli. Perfect correlation was obtained with five antibiotics, whereas norfloxacin, nitrofurantoin and tetracycline were responsible for 13(65%) of the 20 discrepancies. PMID- 11871614 TI - Influence of intestinal anaerobes and organic acids on the growth of enterohaemorrhagic Escherichia coli O157:H7. AB - A suspension of human faeces (FS) and its anaerobic culture (FC), bacterial metabolic products and organic acids were examined for inhibitory effects on growth and verotoxin 2 (VT2) production of Escherichia coli O157:H7 in vitro. FS and FC showed a marked inhibitory activity to growth and production of VT2 by E. coli O157:H7 under anaerobic conditions. They may have both bacteriostatic and bactericidal effects on E. coli O157. The growth of E. coli O157 was markedly suppressed by acetic, propionic and butyric acids compared with hydrochloric acid and lactic acid at concentrations between 25 mM and 40 mM, being proportional to the pH values. At pH 5.5, 40 mM of short-chain fatty acids (SCFAs) almost completely inhibited the growth of E. coli O157. SCFAs markedly inhibited the growth of E. coli O157 at pH 6.0 rather than pH 7.0. Propionic acid is likely to be more suppressive to E. coli than acetic and butyric acids. The production of VT2 was approximately proportional to the growth of E. coli O157. However, incubation for 24 h in vitro showed that the growth and VT2 production of E. coli O157 decreased but were not completely inhibited at pH 6.5 and 7.0 in a mixture of acetic, propionic and butyric acids at a physiological concentration (110 mM, 60:25:25, respectively, in molar ratio). It is probable that the colonic microflora could contribute to a reduction of E. coli O157:H7 infections via the activation of intestinal fermentation by dietary manipulation or something similar to give pH 6.0 or <6.0 and that factors such as age, chemical therapy and body condition, which have effects on the balance of the intestinal microflora, would be associated with the incidence rates of E. coli O157 infections. PMID- 11871615 TI - Characterisation of aerobic gram-negative bacteria from subgingival sites of dogs -potential bite wound pathogens. AB - Ninety-eight aerobic, gram-negative bacterial isolates from subgingival samples from family-owned dogs with naturally occurring periodontitis were characterised phenotypically by conventional biochemical testing, by cellular fatty acid profiling and by the use of commercial identification systems. The majority (48, 81%) of the fermentative isolates but only 18% of the non-fermenters were identified by conventional biochemical testing alone. With additional cellular fatty acid profiling, another 7 (12%) fermentative and 23 (59%) non-fermentative isolates were identified to genus or group level. Cellular fatty acid analysis was essential for the identification of most non-fermenters, many of which are difficult to identify due to a paucity of positive reactions in routine biochemical tests. Commercial identification systems were less useful and did not contribute to further identification of these problematic isolates. This study underlines the difficulties encountered in the identification of canine oral bacteria--a group of potential bite wound pathogens--and presents schemes for microbiology laboratories to characterise such isolates. PMID- 11871616 TI - Comparison of Candida ID medium with sabouraud-chloramphenicol agar for the isolation of yeasts from clinical haematology surveillance specimens. AB - Candida ID is a new chromogenic medium for the identification of yeasts from clinical specimens. C. albicans produces blue pigmentation, whereas pink pigmentation is produced by C. tropicalis, C lusitaniae, C. guilliermondii and C. kefyr; other Candida species appear white. In this study, 240 clinical samples (throat swabs and stool samples) from haematology patients were inoculated on to Candida ID and Sabouraud-chloramphenicol agar in parallel, yielding a total of 105 yeasts; the media had overall detection rates of 85.7% and 86.7% respectively. The sensitivity of Candida ID for identification of C. albicans by blue pigmentation was 52.9% at 24 h and 94.1% at 48 h. Specificity of the blue pigmentation was 100% at 48 h. Two strains of C. tropicalis were identified, one produced pink pigmentation at 72 h, the other strain did not produce any pigmentation after 5 days. Candida ID was superior in detecting mixtures of yeasts compared with Sabouraud-chloramphenicol agar. Candida ID is a suitable primary isolation medium for yeasts from clinical specimens, providing rapid direct identification of C. albicans and enhanced detection of mixtures. PMID- 11871618 TI - Susceptibility of Helicobacter pylori and its urease activity to the peroxidase hydrogen peroxide-thiocyanate antimicrobial system. AB - The susceptibility of Helicobacter pylori to the antimicrobial system involving lactoperoxidase, hydrogen peroxide and thiocyanate was investigated. The inhibitory effect of the system on the urease activity of H. pylori, which plays a role in its colonisation of the stomach, was also investigated. Twelve H. pylori strains examined, including 10 clinical isolates, were all inhibited by the peroxidase system in brain-heart infusion broth supplemented with fetal calf serum, but to different extents. The killing effect was observed within 3 h. Although bacterial viability recovered afterwards, there was still a clear difference between cultures incubated in the presence of the complete system and control cultures incubated in the absence of lactoperoxidase, after incubation for 24 h. The urease activity and viability of H. pylori were both inactivated by this system in phosphate buffer. These effects were dependent on the concentrations of both lactoperoxidase and hydrogen peroxide and were abolished by the addition of cysteine. Furthermore, these effects were observed when bovine lactoperoxidase was replaced by recombinant human lactoperoxidase or native or recombinant human myeloperoxidase. The peroxidase system found in saliva and milk may contribute to the host defence against H. pylori infection and inhibition of transmission via the oral route. PMID- 11871617 TI - Role of interferon-gamma in inflammatory responses in murine respiratory infection with Legionella pneumophila. AB - The role of interferon (IFN)-gamma in host inflammatory responses, including inflammatory cytokine production, in experimental pneumonia with Legionella pneumophila was examined in IFN-gamma knockout (IFN-gamma-/-) mice. IFN-gamma-/- mice and wild-type BALB/cA mice were inoculated intranasally with L. pneumophila strain KC. The survival rate of IFN-gamma-/- mice was significantly lower than that of control mice. Viable bacterial counts in lungs and blood showed a rapid and continuous increase in IFN-gamma-/- mice, in contrast to a gradual decrease in the lungs and an intermittent bacteraemia in control mice. Histopathological analysis of L. pneumophila-infected lung tissues demonstrated mild pneumonia in control mice, whereas severe pneumonia was shown in IFN-gamma-/- mice. During the late stages of infection, the number of total bronchoalveolar lavage (BAL) cells was significantly higher in IFN-gamma-/- than in control mice. The concentrations of tumour necrosis factor-alpha and interleukin-1beta in sera of IFN-gamma-/- mice were significantly lower in control mice during the early stages of infection, suggesting suppressed production of inflammatory cytokines in IFN gamma-/- mice. In contrast, during the late stages of infection, the levels of these cytokines were significantly higher in sera of IFN-gamma-/- mice than in control mice, suggesting severe and systemic infection in IFN-gamma-/- mice. The findings suggest that retardation of host immune responses, including inflammatory cytokine production caused by deficiency of IFN-gamma, might allow the bacteria to grow and cause fulminant pneumonia. PMID- 11871619 TI - Experimental Helicobacter pylori gastric infection in miniature pigs. AB - An experimental Helicobacter pylori infection in miniature pigs was developed and investigated. Eighteen miniature pigs were inoculated with an H. pylori strain that has high virulence in mice at c. 5 x 10(10) cfu. H. pylori infection in miniature pigs was achieved by the administration of agar 1% in brucella broth with fetal bovine serum 10% just before inoculation. The bacterial colonisation and distribution were analysed by mapping of viable cell counts in the stomach in pigs of three different ages. The mapping assay was achieved on post-infection day 3 for the 5-day-old and 2-week-old pigs, and between days 41 and 43 for 3 month-old pigs. The highest cell counts were observed in 5-day-old pigs, which averaged 4.9 x 10(6) cfu/g of mucosa (n = 4). The bacteria were colonised mainly in the cardiac and fundus gland region in the 5-day-old and 2-week-old pigs, whereas the colonisation sites did not depend on the region in the 3-month-old pigs. Biopsy assay of the antral mucosa of a 3-month-old pig after H. pylori infection showed that this infection persisted for >22 months. Serum antibody against H. pylori was detected in the infected pigs but not in the uninfected animal. Immunostaining demonstrated the presence of bacteria on the epithelial surface of the infected pigs. A microscopic finding common to all the infected pigs, focal gastritis with infiltration of lymphocytes detected on the lesser curvature of the stomach, resembled the microscopic appearance in H. pylori infected human patients. These results suggest that miniature pigs might be a suitable model for studying H. pylori infection. PMID- 11871620 TI - A new chromosomal locus associated with gut-modulated phenotypes in Salmonella enterica serotype typhimurium. AB - A cosmid DNA library had been constructed previously from 40-kb fragments of genomic DNA from a virulent invasive strain of Salmonella enterica serotype Typhimurium (TML) in an avirulent hypo-invasive Typhimurium strain (LT7). Selection of invasive clones from the library was attempted by iterative passage through a rabbit ileal organ culture. After the fourth passage, a clone, designated LT7(pHC20-2), was isolated. Exposure to both gut tissue and Caco-2 cells enhanced the growth, invasiveness for gut and Caco-2 cells, and flagellin expression of LT7(pHC20-2) although its invasiveness was less than that of strain TML. Expression of appendages (surface structures c. 60-70 nm diameter) was shown to play a role in but not to confer invasiveness, and was demonstrated in the absence of direct contact with eukaryotic cells. Exposure to gut tissue also affected the expression of several outer-membrane proteins (OMPs) in all four Salmonella strains--TML, LT7, LT7(pHC79), LT7(pHC20 2)--used in this work. As the genes involved in flagella, invasin and porin expression are distributed around the salmonella chromosome, it is possible that pHC20-2 encodes a pleiotropic regulator of genes involved in gastro-enteritic virulence and adaptation to the in-vivo gut environment. pHC20-2 mapped at c. centisome 25 on the salmonella chromosome close to, but distinct from, SPI-5. PMID- 11871621 TI - Epidemiological investigation of a fatal case of cholera in Japan by phenotypic techniques and pulsed-field gel electrophoresis. AB - A fatal case of cholera caused by Vibrio cholerae 01 El Tor serotype Ogawa occurred in Aichi Prefecture, Japan in 1995. The patient was identified locally, but the route of the infection was unknown. The causative isolate and 38 other domestic and imported V. cholerae O1 isolates, obtained between 1984 and 1997, were analysed by prophage typing, antimicrobial susceptibility testing and pulsed field gel electrophoresis (PFGE). This was done to determine whether the isolate from this case differed from others associated with either mild cholera infections or asymptomatic carriage, and to elucidate the route of infection. Cholera toxin (CT) from 37 toxigenic isolates was assayed semi-quantitatively. The 39 isolates were divided into 12 temporary types in accordance with the results of the three typing techniques. The isolate from the fatal infection and nine other isolates were classified as temporary type IV. No difference in CT production was found between the isolate from the fatal case and the other 36 toxigenic isolates. Taken together, it is unlikely that a V. cholerae 01 isolate of distinguishable type was responsible for the fatal illness. Temporary type IV isolates were frequently present in both domestic and imported cases from 1994 to 1997 in Aichi, but they did not emerge before 1993. These results suggest that a new clone was introduced after 1993 from overseas and then disseminated into Aichi, and this may have been an important step in triggering the fatal case of cholera. PMID- 11871622 TI - Fusobacterium necrophorum infections in England and Wales 1990-2000. AB - In response to a marked increase in both the number of Fusobacterium necrophorum bacteraemia reports to the PHLS Communicable Disease Surveillance Centre and the number of F. necrophorum isolates referred to the PHLS Anaerobe Reference Unit in 1999, the data from both sources on F. necrophorum infections were reviewed for the decade 1990-2000. There were 208 reports of F. necrophorum bacteraemia (average 19/year; range 14-34/year) with a peak in incidence in the late winter months; 68% were from male patients and the peak age range was 16-23 years. Of 205 referred isolates of F. necrophorum, 122 (59%) were from blood cultures and these represented 58% of the bacteraemia reports; the others were from brain and soft tissue abscesses, pleural and joint fluids, eyes, ears and lymphatic tissue. The average number of referrals was 19/year (range 9-37/year). The peak year for bacteraemia reports (34) and isolate referrals (37) was 1999; this increase was not sustained in 2000. All isolates were susceptible to metronidazole, but 2% were resistant to penicillin and 15% to erythromycin. F. necrophorum continues to be a regular but uncommon cause of bacteraemia and metastatic abscesses following an acute sore throat, especially in young, otherwise healthy adults. PMID- 11871623 TI - Molecular epidemiology of tuberculosis in Elche, Spain: a 7-year study. AB - The epidemiology of tuberculosis in Elche (Spain) was studied by restriction fragment-length polymorphism (RFLP) typing of clinical isolates of Mycobacterium tuberculosis over a 7-year period. A total of 165 isolates was typed and the clinico-epidemiological data of the patients were studied retrospectively. An overall cluster aggregation of 52.4% was found, rising to 71.43% in HIV-positive patients. There was greater aggregation in younger patients, but no statistically significant differences when other variables were analysed. The percentage of aggregation was higher than in other studies in Spain and this may be due to the longer time period of this study. The high percentage of aggregation in young patients and in thosewho were HIV-positive suggests increased recent transmission in both groups. PMID- 11871624 TI - Bacillus cereus cellulitis from contaminated heroin. AB - Concern exists over recent unexplained deaths among intravenous drug users. This report describes a patient with crepitant cellulitis who was admitted complaining of severe pain in the right forearm. Ultrasonography demonstrated gas in the tissues and he was referred for early surgical debridement of the arm. He was treated with intravenous benzyl penicillin, gentamicin and metronidazole and made a full recovery. Aspirate samples grew Bacillus cereus, morphologically similar to the isolate obtained from a sample of the patient's own heroin. Antibiogram and API 50CHB profiles were also similar. Further typing included 'H' flagellar serotyping, which found both blood and heroin strains to be non-typable, and amplified fragment polymorphism analysis, which showed that the strains were indistinguishable. Genotyping of two selected genes from B. cereus confirmed almost certain identity between the two strains. This case illustrates the potential virulence of B. cereus when inoculated into tissues, and to our knowledge, is the first report to demonstrate a conclusive microbiological link between contaminated heroin and serious sepsis in a drug user due to B. cereus. PMID- 11871626 TI - Factors associated with nurses' poor knowledge and practice of transfusion safety procedures in Aquitaine, France. AB - OBJECTIVES: To (1) describe knowledge, attitudes, and reported practice of blood transfusion of nurses in Aquitaine's hospitals; (2) measure the potential threat for patient safety of poor transfusion-related knowledge and practice; and (3) identify factors associated with poor knowledge and practice. DESIGN: Survey conducted in 14 hospitals in Aquitaine (one university and 13 general hospitals). SETTING: Hospitalized care. PARTICIPANTS: A random sample of nurses. MATERIALS: Data were collected anonymously by investigators through structured individual interviews. The questionnaire contained mainly knowledge and practice questions about blood transfusion regulation. MAIN OUTCOME MEASURES: Hazardous knowledge and practice scores have been constructed, reflecting the levels of potential danger in the answers to the questionnaire. Factors associated with these scores have been studied using a random-effect linear regression. RESULTS: In our sample of 1090 nurses, poor knowledge and practice concerned mainly (1) the bedside blood compatibility test [proportion of responses (PR) with potential life threat between 12.7 and 35.5%]; (2) pre-transfusion compatibility check when receiving blood units (PR = 34.5%); (3) delay between screening of red cell antibodies and transfusion (PR = 20.5%); (4) delay in preservation of blood unit in the ward (PR = 33.4%); and (5) recognition of abnormal reactions after transfusion (PR = 47.1%). Frequency of transfusion and training were the factors most strongly associated with hazardous knowledge and practice scores. CONCLUSION: Low training and transfusion activity were key determinants of poor transfusion-related knowledge and practice. PMID- 11871625 TI - Mortality associated with the quality of care of patients hospitalized with congestive heart failure. AB - OBJECTIVE: This study examined the association between use of angiotensin converting enzyme inhibitors (ACEIs) and risk of death in elderly patients hospitalized with left ventricular systolic dysfunction (LVSD). DESIGN: Retrospective cohort study. SETTING: Despite evidence showing the benefit of treating LVSD with ACEI, elderly patients with LVSD are often not treated with an ACEI. Concern that the risk of ACEI treatment might exceed the benefits in elderly patients is a possible reason. STUDY PARTICIPANTS: We abstracted records from 2943 Medicare beneficiaries hospitalized for congestive heart failure in 69 hospitals in five states. The presence of LVSD was determined from recorded ejection fractions or a narrative description of ventricular function. Discharge medications and dosages were abstracted. MAIN OUTCOME MEASURES: Mortality was tracked for one year using Health Care Finance Administration MEDPRO files. RESULTS: There were 621 patients aged 65 years or older with LVSD. The mean age (SD) was 77.4 years (7.0). At discharge 79% were prescribed an ACEI and, of these, 47% were discharged at the dose recommended by clinical practice guidelines. There were 195 deaths (31.4%) during the year of follow-up. Compared with patients discharged at a recommended ACEI dose, patients not prescribed an ACEI at discharge had an adjusted hazard ratio for death (95% CI) of 1.63 (1.02, 2.60) and patients prescribed an ACEI at a less than recommended dose had a hazard ratio of 1.30 (0.86, 1.97). CONCLUSIONS: Our results show that ACEI use at discharge in elderly patients with LVSD is associated with decreased risk of death. PMID- 11871627 TI - Structural and process quality in the management of diabetic emergencies in Germany. AB - OBJECTIVE: Although acute complications of diabetes account for approximately 3% of all emergency calls, clinically relevant indicators of structural and process quality in the management of diabetic emergencies have not yet been studied. The purpose of this investigation was, therefore, to collect representative data on these indicators for the whole of Germany. METHODS: Standardized questionnaires comprising 20 items were sent to all 312 emergency medical services in Germany. Apart from demographic data, information was obtained about the diagnostic materials and drugs carried by the ambulances, methods of blood glucose measurement, the level of qualification of the emergency teams, the frequency of diabetic emergencies, and the need for further training. RESULTS: The return rate of the questionnaires was 55%, corresponding to 172 emergency medical service districts serving a total population of 45.3 million. The data revealed deficits with regard to structural and process quality. Thus, only 6% of ambulances carried glucagon and only 11% ketone test strips. In 57% capillary blood was used for glucose determination, in 17% visually read test strips were still used. While in some districts hospital admission after hypoglycaemic episodes was mandatory even for patients well educated about their diabetes, in other districts multimorbid patients on oral antidiabetics were sometimes only treated at the emergency scene. Emergency medical technicians increasingly carried out both the diagnosis and treatment of diabetic emergencies. CONCLUSIONS: The structural and process quality of the management of diabetic emergencies in Germany is in need of improvement. The most important factor is continuing education of the entire emergency team. PMID- 11871628 TI - Using quality improvement strategies to enhance pediatric pain assessment. AB - OBJECTIVE: To evaluate the impact of a quality improvement approach to implementing developmentally appropriate pain assessment guidelines for pediatric inpatients. Patient and staff satisfaction with pain assessment and management, and staff documentation were evaluated one year following the implementation of the revised pain assessment guidelines. DESIGN: Quasi-experimental design. SETTING: The pediatric hematology/oncology unit of a regional children's hospital. Study participants. A convenience sample of 36 children and 68 staff at time 1 (TI) and 49 children and 82 staff at time 2 (T2). INTERVENTIONS: Staff were educated on the use of pediatric pain assessment tools and a standardized pain assessment protocol was put into practice. Children or their primary caregivers were interviewed, using a questionnaire modified from the American Pain Society quality assurance guidelines, regarding their experiences with pain assessment and management on the unit at T1, just prior to the staff education, and one year later at T2. Multidisciplinary unit staff completed a parallel questionnaire at T1 and T2. MAIN OUTCOME MEASURES: Patient and staff satisfaction with pain assessment and management, and chart audit of compliance with pain assessment documentation. RESULTS: Patients, family members, and staff reported increased pain assessment, improved staff responsiveness to patients' pain and greater satisfaction with assessment tools between TI and T2. Increased compliance with the assessment guidelines was confirmed by chart audit. CONCLUSIONS: Improved pain assessment and management practices with resultant increase in patient and staff satisfaction can be achieved and sustained over time using quality improvement strategies. PMID- 11871629 TI - Measuring job satisfaction in residential aged care. AB - BACKGROUND: Staff satisfaction has received increasing recognition as an important factor influencing service quality and in particular the quality of residents' lives in residential aged care facilities, where staff typically have a long-term and close relationship with residents. Consequently, a valid and reliable instrument is required to assess staff satisfaction in this particular context. OBJECTIVE: This paper aims to assess the factor structure, reliability, and validity of the Measure of Job Satisfaction (MJS) instrument when used in residential aged care facilities. DESIGN: A cross-sectional survey design was used to collect the required information, and a stratified random sampling method was utilized to select facilities. Exploratory and confirmatory factor analyses were conducted to assess the factor structure of staff satisfaction via the MJS. SETTING: Both high and low care residential aged care facilities in Western Australia. STUDY PARTICIPANTS: Nine hundred and eighty-three staff (including the Director of Nursing, manager, registered nurses, enrolled nurses, nursing assistants, and therapists) in 70 residential aged care facilities. RESULTS: An acceptable five-factor (22-item) measurement model was derived. The Cronbach's alpha reliability levels range from 0.86 to 0.95. Convergent and discriminant validity are also satisfactory. CONCLUSION: This investigation has confirmed that a modified MJS is a reliable and valid instrument for assessing staff satisfaction in residential aged care settings. PMID- 11871631 TI - Predictors of short-term clinical response to acute asthma care in adults. AB - OBJECTIVE: We assessed the effects of potential patient risk factors on short term clinical response to acute asthma care among adults who often require emergency department (ED) visits and hospitalizations to manage their asthma. DESIGN AND SETTING: Prospective cohort study that included adult patients treated for acute asthma, which was conducted in a US public hospital ED between March 1997 and August 1999, with a 2- to 3-week follow-up. STUDY PARTICIPANTS: Three hundred and nine patients completed the study. MAIN MEASURES: We identified patient risk factors that predicted lower peak expiratory flow rate (PEFR) change over 2-3 weeks following acute asthma care. Potential risk factors were ozone exposure, indoor allergy and exposure, smoking, upper respiratory infection in the last month, lower asthma knowledge, and medication non-adherence. RESULTS: Univariate analyses indicated that lower asthma knowledge significantly and positively correlated with lower PEFR change (r = 0.15, P = 0.01). Multivariate analysis that controlled for patient case-mix indicated that indoor allergy and exposure [b = 32.76, 95% confidence interval (CI) = 3.98-61.53, P = 0.03] significantly predicted lower PEFR change. There was no change in the multivariate analysis when the absence of treatment with corticosteroids during the 2-3 weeks before follow-up was added as a potential patient risk factor. CONCLUSION: The study suggests that interventions are needed to target the patient risk factors, indoor allergen exposure, and poor asthma knowledge, to promote short-term clinical response to acute asthma care in adults, especially among economically disadvantaged inner-city residents. PMID- 11871630 TI - Hospital staffing, organization, and quality of care: cross-national findings. AB - OBJECTIVE: To examine the effects of nurse staffing and organizational support for nursing care on nurses' dissatisfaction with their jobs, nurse burnout, and nurse reports of quality of patient care in an international sample of hospitals. DESIGN: Multisite cross-sectional survey. SETTING: Adult acute-care hospitals in the United States (Pennsylvania), Canada (Ontario and British Columbia), England, and Scotland. STUDY PARTICIPANTS: 10 319 nurses working on medical and surgical units in 303 hospitals across the five jurisdictions. INTERVENTIONS: None. MAIN OUTCOME MEASURES: Nurse job dissatisfaction, burnout, and nurse-rated quality of care. RESULTS: Dissatisfaction, burnout, and concerns about quality of care were common among hospital nurses in all five sites. Organizational/managerial support for nursing had a pronounced effect on nurse dissatisfaction and burnout, and both organizational support for nursing and nurse staffing were directly, and independently, related to nurse-assessed quality of care. Multivariate results imply that nurse reports of low quality care were three times as likely in hospitals with low staffing and support for nurses as in hospitals with high staffing and support. CONCLUSION: Adequate nurse staffing and organizational/managerial support for nursing are key to improving the quality of patient care, to diminishing nurse job dissatisfaction and burnout and, ultimately, to improving the nurse retention problem in hospital settings. PMID- 11871632 TI - Flawed communication systems result in patient harm. PMID- 11871633 TI - Adverse cutaneous reactions to drugs. PMID- 11871634 TI - Management of lower leg cellulitis. PMID- 11871635 TI - Chronic lower limb oedema. PMID- 11871636 TI - Hospital episode statistics: time for clinicians to get involved? AB - Hospital episode statistics contain clinical data. They are used for many purposes, including monitoring activity in the NHS and the allocation of funds. More recently they have been applied to monitoring performance, and it is intended that they will inform consultant appraisal and revalidation. The validity of hospital episode statistics was questioned by Korner in 1982. Recent publications have shown that problems persist in England and Wales, and that the quality of the data is inadequate for the task. The lack of involvement of clinicians in the process of data collection and validation is no longer acceptable. To rectify the situation there should be a change of process and culture, supported by education and investment. NHS data definitions of terms such as 'spells', 'episodes' and 'diagnoses' need to be reviewed. The development of separate data processes to monitor national service frameworks is regrettable. PMID- 11871637 TI - Consultant nurses and their potential impact upon health care delivery. AB - The role of the consultant nurse should be fully accepted by medical and nursing staff in order to develop new ways of working, delivering cost-effective quality patient care and meeting the agenda set out in the NHS plan. The involvement of the medical staff in identifying need, ensuring appropriate funding and subsequent training, and developing consultant-nurse posts is vital to their success. Several different healthcare professionals have or can attain the competencies that historically were considered to be the private domain of another profession. PMID- 11871638 TI - Shift work for specialist registrars in acute medicine: more questions than solutions. AB - Trainees' hours are under review once again, due to the changes incurred by the latest pay deal and the likely enforcement of the European Working Time Directive. Until now, specialist registrars (SpRs) training in general internal medicine (GIM) have been accustomed to residential on-call rotas or partial shift schemes as a means of covering emergency work. Some NHS trusts have suggested that GIM SpRs should now work full shifts to meet the restrictions of European Working Time Directive. This article outlines the views of the Royal College of Physicians' Trainees Committee on the potential impact of full shift work for GIM SpRs on medical services. Impaired continuity of inpatient care and reduced staffing levels for outpatient services were felt to be the most serious possible consequences. Some short-term solutions are suggested, but in the long term, increased numbers of both consultants and SpRs will be vital to maintain and improve standards in acute medicine. PMID- 11871640 TI - Consultant nurses. PMID- 11871639 TI - Potential therapeutic targets in the rapidly expanding field of purinergic signalling. AB - The concept of a purinergic signalling system, using purine nucleotides and nucleosides as extracellular messengers, was first proposed over 30 years ago. After a brief historical review and update of purinoceptor subtypes, this article focuses on the diverse physiological roles of adenosine triphosphate, adenosine diphosphate, uridine triphosphate and adenosine. These molecules mediate short term (acute) signalling functions in neurotransmission, secretion and vasodilation, and long-term (chronic) signalling functions in development, regeneration, proliferation and cell death. Plasticity of purinoceptor expression in pathological conditions is frequently observed, including an increase in the purinergic component of parasympathetic nervous control of the human bladder in interstitial cystitis and outflow obstruction, and in sympathetic cotransmitter control of blood vessels in hypertensive rats. The antithrombotic action of clopidogrel (Plavix), a P2Y12 receptor antagonist, has been shown to be particularly promising in the prevention of recurrent strokes and heart attacks in recent clinical trials (CAPRIE and CURE). The role of P2X3 receptors in nociception and a new hypothesis concerning purinergic mechanosensory transduction in visceral pain will be considered, as will the therapeutic potential of purinergic agonists or antagonists for the treatment of supraventricular tachycardia, cancer, dry eye, bladder hyperactivity, erectile dysfunction, osteoporosis, diabetes, gut motility and vascular disorders. PMID- 11871641 TI - Brain attack: a new approach to stroke. AB - 'Brain' attack' is a new term to describe the acute presentation of stroke which emphasises the need for urgent action. The article describes the basis for this new approach to acute stroke treatment. Rational treatment requires individual causes of stroke to be identified early and treatment targeted at the mechanism. Acute stroke treatment aims to preserve the ischaemic penumbra, protect neurons against further ischaemia and enhance brain plasticity to maximise recovery. There is a strong evidence base supporting the routine use of aspirin, but not heparin, in acute ischaemic stroke. There is also convincing evidence supporting intravenous thrombolysis using recombinant tissue plasminogen activator in selected patients within 3 hours of stroke onset. Surprisingly, as many as 33% of suspected-stroke patients arrive in Accident & Emergency departments in the UK within 3 hours of onset. New techniques in MR imaging, particularly diffusion weighted imaging, are transforming the approach to diagnosis of acute stroke. Although neuroprotective drugs have proved disappointing, active neuroprotection in acute stroke should include control of blood pressure within certain limits, antipyretic therapy, maintenance of blood glucose, and early feeding and fluid replacement. Surgical hemicraniectomy should be considered in patients with malignant cerebral oedema. There is good evidence that the best way to enhance recovery from stroke is to admit the patient to a stroke unit. To enable patients to benefit from the early active approach outlined in the article, the following are needed: the development of acute stroke units; imaging protocols; and education of patients, general practitioners and the ambulance services. Stroke care has become a specialised field, requiring input from stroke physicians, as well as the multidisciplinary rehabilitation team. The British Association of Stroke Physicians (BASP) has therefore developed a curriculum which is designed to lead to the development of a new sub-specialty of stroke medicine. PMID- 11871642 TI - Imaging the neurochemical brain in health and disease. AB - Neurochemical transmission is a fundamental element of brain organisation that has been relatively unexplored in the living human brain. Continuing advances in radionuclide imaging, particularly positron emission tomography (PET) and single photon emission tomography (SPET), mean that elements of neurochemical transmission can now be directly measured in vivo. With these techniques convincing abnormalities of monoaminergic neurotransmitter systems have been revealed in illnesses such as Parkinson's disease and schizophrenia. Furthermore, mechanisms of drug action and treatment responses can be monitored in vivo. This brief review describes some of our recent attempts to image the neurochemical brain in health and disease at the MRC Cyclotron Unit, Hammersmith Hospital, London. PMID- 11871643 TI - Federation of the Royal Colleges of Physicians: census 2000. PMID- 11871644 TI - Medieval uroscopy and its representation on misericords--part 2: misericords. AB - By the fifteenth century the practice of uroscopy was falling into disrepute and the uroscopy flask (matula) became a symbol of ridicule. On the carved misericords in choir stalls, the physician holding the matula was commonly represented as an ape, with the allegorical implications of foolishness, vanity and even lechery. The ape uroscopist was frequently shown with his friend the fox, an animal that was often used to satirise the less-than-perfect cleric, and this association may reflect the close ties between the medical and clerical professions in the medieval period. PMID- 11871645 TI - Cardiopulmonary resuscitation and do not attempt resuscitation orders: legislation may be helpful. PMID- 11871646 TI - Cardiopulmonary resuscitation and do not attempt resuscitation and do not attempt resuscitation orders: legislation may be helpful. PMID- 11871648 TI - Clinical aviation medicine: safe travel by air. PMID- 11871647 TI - Biological weapons: the facts not the fiction. PMID- 11871649 TI - Autoimmune haematological disorders. PMID- 11871650 TI - A crisis of identity. PMID- 11871651 TI - Modification and modernization of professional regulation. PMID- 11871657 TI - Human ribonuclease P: subunits, function, and intranuclear localization. AB - Catalytic complexes of nuclear ribonuclease P (RNase P) ribonucleoproteins are composed of several protein subunits that appear to have specific roles in enzyme function in tRNA processing. This review describes recent progress made in the characterization of human RNase P, its relationship with the ribosomal RNA processing ribonucleoprotein RNase MRP, and the unexpected evolutionary conservation of its subunits. A new model for the biosynthesis of human RNase P is presented, in which this process is dynamic, transcription-dependent, and implicates functionally distinct nuclear compartments in tRNA biogenesis. PMID- 11871658 TI - Influence of the stacking potential of the base 3' of tandem shift codons on -1 ribosomal frameshifting used for gene expression. AB - Translating ribosomes can shift reading frame at specific sites with high efficiency for gene expression purposes. The most common type of shift to the -1 frame involves a tandem realignment of two anticodons from pairing with mRNA sequence of the form X XXY YYZ to XXX YYY Z where the spaces indicate the reading frame. The predominant -1 shift site of this type in eubacteria is A AAA AAG. The present work shows that in Escherichia coli the identity of the 6 nt 3' of this sequence can be responsible for a 14-fold variation in frameshift frequency. The first 3' nucleotide has the primary effect, with, in order of decreasing efficiency, U > C > A > G. This effect is independent of other stimulators of frameshifting. It is detected with other X XXA AAG sequences, but not with several other heptameric -1 shift sites. Pairing of E. coli tRNALYS with AAG is especially weak at the third codon position. We propose that strong stacking of purines 3' of AAG stabilizes pairing of tRNALys, diminishing the chance of codon:anticodon dissociation that is a prerequisite for the realignment involved in frameshifting. PMID- 11871659 TI - Structure of the histone mRNA hairpin required for cell cycle regulation of histone gene expression. AB - Expression of replication-dependent histone genes requires a conserved hairpin RNA element in the 3' untranslated regions of poly(A)-less histone mRNAs. The 3' hairpin element is recognized by the hairpin-binding protein or stem-loop-binding protein (HBP/SLBP). This protein-RNA interaction is important for the endonucleolytic cleavage generating the mature mRNA 3' end. The 3' hairpin and presumably HBP/SLBP are also required for nucleocytoplasmic transport, translation, and stability of histone mRNAs. RNA 3' processing and mRNA stability are both regulated during the cell cycle. Here, we have determined the three dimensional structure of a 24-mer RNA comprising a mammalian histone RNA hairpin using heteronuclear multidimensional NMR spectroscopy. The hairpin adopts a novel UUUC tetraloop conformation that is stabilized by base stacking involving the first and third loop uridines and a closing U-A base pair, and by hydrogen bonding between the first and third uridines in the tetraloop. The HBP interaction of hairpin RNA variants was analyzed in band shift experiments. Particularly important interactions for HBP recognition are mediated by the closing U-A base pair and the first and third loop uridines, whose Watson-Crick functional groups are exposed towards the major groove of the RNA hairpin. The results obtained provide novel structural insight into the interaction of the histone 3' hairpin with HBP, and thus the regulation of histone mRNA metabolism. PMID- 11871660 TI - Differential localization of nuclear-encoded tRNAs between the cytosol and mitochondrion in Leishmania tarentolae. AB - All mitochondrial tRNAs of the kinetoplastid protozoan Leishmania tarentolae are encoded in the nucleus and are imported from the cytosol into the mitochondrion. We previously reported the partitioning of five tRNAs and found that all were shared between the two compartments to different extents. To increase our knowledge of the tRNAs of this organism, and to attempt to understand the signals involved in their subcellular localization, a method to RT-PCR amplify new tRNAs was developed. Various tRNAs were 3' polyadenylated and reverse transcribed with a sequence-tagged primer. The cDNA was tagged by ligation to an anchor oligonucleotide, and the resulting double-tagged cDNA was amplified by PCR. Four new tRNAs were obtained, bringing to 20 the total number of L. tarentolae tRNAs identified to date. The subcellular localization of 17 tRNAs was quantitatively analyzed by two-dimensional gel electrophoresis and northern hybridization. In general, the previously suggested operational classification of tRNAs into three groups (mainly cytosolic, mainly mitochondrial, and shared between the two compartments) is still valid, but the relative abundance of each tRNA in the cytosol or mitochondrion varied greatly as did the level of expression. PMID- 11871661 TI - Identification of domains in apobec-1 complementation factor required for RNA binding and apolipoprotein-B mRNA editing. AB - The C-to-U editing of apolipoprotein-B (apo-B) mRNA is catalyzed by an enzyme complex that recognizes an 11-nt mooring sequence downstream of the editing site. A minimal holoenzyme that edits apo-B mRNA in vitro has been defined. This complex contains apobec-1, the catalytic subunit, and apobec-1 complementation factor (ACF), the RNA-binding subunit that binds to the mooring sequence. Here, we show that ACF binds with high affinity to single-stranded but not double stranded apo-B mRNA. ACF contains three nonidentical RNA recognition motifs (RRM) and a unique C-terminal auxiliary domain. In many multi-RRM proteins, the RRMs mediate RNA binding and an auxiliary domain functions in protein-protein interactions. Here we show that ACF does not fit this simple model. Based on deletion mutagenesis, the RRMs in ACF are necessary but not sufficient for binding to apo-B mRNA. Amino acids in the pre-RRM region are required for complementing activity and RNA binding, but not for interaction with apobec-1. The C-terminal 196 amino acids are not absolutely essential for function. However, further deletion of an RG-rich region from the auxiliary domain abolished complementing activity, RNA binding, and apobec-1 interaction. The auxiliary domain alone did not bind apobec-1. Although all three RRMs are required for complementing activity and apobec-1 interaction, the individual motifs contribute differently to RNA binding. Point mutations in RRM1 or RRM2 decreased the Kd for apo-B mRNA by two orders of magnitude whereas mutations in RRM3 reduced binding affinity 13-fold. The pairwise expression of RRM1 with RRM2 or RRM3 resulted in moderate affinity binding. PMID- 11871662 TI - NMR structure and dynamics of the RNA-binding site for the histone mRNA stem-loop binding protein. AB - The 3' end of replication-dependent histone mRNAs terminate in a conserved sequence containing a stem-loop. This 26-nt sequence is the binding site for a protein, stem-loop binding protein (SLBP), that is involved in multiple aspects of histone mRNA metabolism and regulation. We have determined the structure of the 26-nt sequence by multidimensional NMR spectroscopy. There is a 16-nt stem loop motif, with a conserved 6-bp stem and a 4-nt loop. The loop is closed by a conserved U.A base pair that terminates the canonical A-form stem. The pyrimidine rich 4-nt loop, UUUC, is well organized with the three uridines stacking on the helix, and the fourth base extending across the major groove into the solvent. The flanking nucleotides at the base of the hairpin stem do not assume a unique conformation, despite the fact that the 5' flanking nucleotides are a critical component of the SLBP binding site. PMID- 11871664 TI - Select committee identifies 'key issues' on FMD. PMID- 11871663 TI - RNase E plays an essential role in the maturation of Escherichia coli tRNA precursors. AB - Conversion of tRNA precursors to their mature forms requires the action of both endo- and exoribonucleases. Although studies over many years identified the endoribonuclease, RNase P, and several exoribonucleases as the enzymes responsible for generating the mature 5' and 3' termini, respectively, of Escherichia coli tRNAs, relatively little is known about how tRNAs are separated from long multimeric or multifunction transcripts, or from long leader and trailer sequences. To examine this question, the tRNA products that accumulate in mutant strains devoid of multiple exoribonucleases plus one or several endoribonucleases were analyzed by northern analysis. We find that the multifunction tyrT transcript, which contains two tRNA(Tyr)1 sequences separated by a 209-nt spacer region plus a downstream mRNA, is cleaved at three sites in the spacer region by the endoribonuclease, RNase E. When both RNase E and RNase P are absent, a product containing both tRNAs accumulates. Two multimeric tRNA transcripts, those for tRNA Arg-His-Leu-Pro and tRNA Gly-Cys-Leu also require RNase E for maturation. For the former transcript, products with long 3' extensions on tRNA(Arg), tRNA(His), and tRNA(Pro), as well as the primary transcript, accumulate in the absence of RNase E. For the latter transcript, RNase E cleaves downstream of each tRNA. Little processing of either multimeric transcript occurs in the absence of both RNase E and RNase P. These data indicate that RNase E is a major contributor to the initial processing of E. coli tRNA transcripts, providing substrates for final maturation by RNase P and the 3' exoribonucleases. Based on this new information, a detailed model for tRNA maturation is proposed. PMID- 11871665 TI - Veterinary Products Committee working group report on feline and canine vaccination. AB - * The working group was set up by the Veterinary Products Committee in response to current concern in both the public domain and in the scientific community about possible health risks related to the routine vaccination of cats and dogs. The working group concluded that vaccination plays a very valuable role in the prevention and control of the major infectious diseases in cats and dogs. Although adverse reactions to vaccination, including lack of efficacy, occasionally occur, the working group concluded that the overall risk/benefit analysis strongly supports their continued use. * Although for some diseases there is evidence of a longer duration of immunity following vaccination than the one year which is typically recommended on the product literature, there is currently insufficient information to propose revaccination intervals other than those proposed by the manufacturer and approved by the regulatory process. * Notwithstanding this, in view of the occasional occurrence of adverse reactions, the working group recommends that the product literature indicates that the regime for booster vaccinations is based on a minimum duration of immunity rather than a maximum. The working group further recommends that the product literature should state that a risk/benefit assessment should be made for each individual animal by the veterinary surgeon in consultation with the owner with respect to the necessity for each vaccine and the frequency of its use. * The evidence suggests that cats appear to be susceptible to the occasional development of sarcomas at sites of injection and there is some further evidence to suggest that, although other products may be involved, this may be more associated with the use of vaccines containing aluminium-based adjuvants. The working group therefore recommends that a generic warning to this effect should appear on the product literature for all feline vaccines administered by injection. The working group also highlighted the need for professional and educational bodies in the UK to bring to the attention of veterinary practitioners appropriate methods for prevention, diagnosis and treatment of this serious condition. * The working group considered in depth the monitoring of adverse reactions, including the advantages and disadvantages of surveillance schemes. A range of options for carrying out further epidemiological (analytical) studies was also considered. However, the working group emphasised that surveillance schemes, and the UK Veterinary Medicines Directorate (VMD) Suspected Adverse Reaction (SAR) Surveillance Scheme in particular, provided a very valuable resource. The large database within the VMD scheme (collected since 1985) was analysed as part of this report. Figures were derived in terms of incidence (reporting rate) of certain clinical signs per 10,000 doses, and risk factors as identified by statistical analysis. However, due to a number of constraints, the analysis was not fully comprehensive and the interaction of possible risk factors was not determined. * Product-related control charts were developed in order to detect changes in incidence rates of adverse reactions (per 10,000 doses sold) both within and between different vaccines. Such charts provide a powerful way to detect changing trends in incidence and, when used in conjunction with product characteristics, they may identify possible causes. In general, the data showed that the incidence of adverse reactions to cat and dog vaccines per 10,000 doses of product sold was relatively low. Although under-reporting is a feature of such surveillance schemes, it does appear that, overall, vaccination of cats and dogs should be considered safe and effective. * Finally, the working group was conscious, while preparing this report, of the extensive media coverage that has been given to the issue of the safety of human vaccines, in particular the mumps, measles and rubella (MMR) vaccine. The working group emphasises that the conclusions and recommendations included in this report relate only to the vaccines used in cats and dogs. The issues identified are specific to the diseases and species examined and no attempt should be made to draw analogous conclusions in relation to vaccines administered to humans. PMID- 11871666 TI - Risk factors for Johne's disease in Scotland--the results of a survey of farmers. AB - The reported incidence of Johne's disease has been increasing in the east of Scotland since 1993. A postal questionnaire survey was sent to 127 farms to identify potential risk factors for Johne's disease in relation to wildlife and farm management practices, and 86 returns were obtained. Of 22 farms which had been assumed to be free of the disease, on the basis of information held by local veterinary centres, seven (32 per cent) reported cases of Johne's disease in the 1990s, indicating that the disease is under-reported. Logistic regression analyses showed that eight of 63 potentially explanatory variables were significant at the 5 per cent level in affecting the likelihood of farms reporting Johne's disease. Of these, large numbers of livestock and rabbits, and access of wildlife to feed stores were the clearest and most consistent risk factors associated with the disease. The application of manure to grazing pasture, the type of water supply for the cattle and the numbers of crows were also related to the presence of Johne's disease but the nature of these relationships was less clear. Only 38 per cent of the farms reported taking any control measures to combat Johne's disease, but three of the control measures were relevant to risk factors identified as significant by the survey, namely maintaining a clean water supply, controlling rabbits and not spreading manure on to grazing pasture. PMID- 11871667 TI - Infectious agents identified in pigs with multifocal interstitial nephritis at slaughter. AB - One kidney was taken from each of 100 pigs at slaughter; 50 had gross lesions of multifocal interstitial nephritis and 50 had no gross lesions. Forty-nine of the affected kidneys had lesions that were characterised by the presence of either a few randomly distributed or numerous widely disseminated pale foci, 1 to 3 mm in diameter, on the cortical surface (white-dotted kidneys). Microscopically, these focal inflammatory lesions often had a distinct lymphofollicular pattern (follicular nephritis). Lesions of chronic vasculitis were observed in 21 of the affected kidneys. Histologically, the control kidneys had only small and sparse inflammatory foci. Standard bacterial cultures of kidneys of both groups were not significant, and cultures for the isolation of leptospires were all negative. Virological examination of the kidney homogenates by PCR did not reveal any porcine reproductive and respiratory syndrome virus and only a few cases were positive for the porcine circovirus type 1. However, porcine parvovirus (PPV) and porcine circovirus type 2 (PCV-2) were detected in many kidneys of both groups but in a significantly higher proportion of the kidneys with interstitial nephritis. There was a significant association between the lesions and the presence of PPV and PCV-2 with odds ratios of 7.5 (P<0.0001) and 3.4 (P=0.0074), respectively, and the odds ratio increased to 22.7 (P<0.0001) when both viruses were identified in the same kidney. However, a subsample of kidneys taken from both groups were negative by immunohistochemistry for the presence of PPV and PCV 2 antigens. PMID- 11871668 TI - Fusion of the distal sesamoid and distal phalanx in a yearling colt. AB - This paper describes a rare malformation of the distal portion of the left foredigit of a 15-month-old half-bred colt which was severely lame. Radiological, pathological and tomographic studies revealed hypoplasia of the metacarpophalangeal skeleton, absence of the distal sesamoid and deformation of the distal phalanx. Morphological analysis of the lesion and computerised measurement of the width of the articular surface of the distal phalanx suggested an early fusion between the distal sesamoid and distal phalanx. PMID- 11871669 TI - Ultrasonography in the differential diagnosis of azotaemia in an adult Cheviot ram. PMID- 11871670 TI - Secondary vaginal pouch in a bitch. PMID- 11871671 TI - Duty of care regarding night visits. PMID- 11871672 TI - Release into the wild of grey squirrels. PMID- 11871673 TI - Vaccination and FMD. PMID- 11871674 TI - Reducing headache disability in children and adolescents. PMID- 11871675 TI - Acute rhinosinusitis. PMID- 11871676 TI - Hyperbilirubinemia in the term newborn. AB - Hyperbilirubinemia is one of the most common problems encountered in term newborns. Historically, management guidelines were derived from studies on bilirubin toxicity in infants with hemolytic disease. More recent recommendations support the use of less intensive therapy in healthy term newborns with jaundice. Phototherapy should be instituted when the total serum bilirubin level is at or above 15 mg per dL (257 micromol per L) in infants 25 to 48 hours old, 18 mg per dL (308 micromol per L) in infants 49 to 72 hours old, and 20 mg per dL (342 micromol per L) in infants older than 72 hours. Few term newborns with hyperbilirubinemia have serious underlying pathology. Jaundice is considered pathologic if it presents within the first 24 hours after birth, the total serum bilirubin level rises by more than 5 mg per dL (86 micromol per L) per day or is higher than 17 mg per dL (290 micromol per L), or an infant has signs and symptoms suggestive of serious illness. The management goals are to exclude pathologic causes of hyperbilirubinemia and initiate treatment to prevent bilirubin neurotoxicity. PMID- 11871677 TI - Information from your family doctor. Jaundice and your baby. PMID- 11871678 TI - Burning mouth syndrome. AB - Burning mouth syndrome is characterized by a burning sensation in the tongue or other oral sites, usually in the absence of clinical and laboratory findings. Affected patients often present with multiple oral complaints, including burning, dryness and taste alterations. Burning mouth complaints are reported more often in women, especially after menopause. Typically, patients awaken without pain but note increasing symptoms through the day and into the evening. Conditions that have been reported in association with burning mouth syndrome include chronic anxiety or depression, various nutritional deficiencies, type 2 diabetes (formerly known as non-insulin-dependent diabetes) and changes in salivary function. However, these conditions have not been consistently linked with the syndrome, and their treatment has had little impact on burning mouth symptoms. Recent studies have pointed to dysfunction of several cranial nerves associated with taste sensation as a possible cause of burning mouth syndrome. Given in low dosages, benzodiazepines, tricyclic antidepressants or anticonvulsants may be effective in patients with burning mouth syndrome. Topical capsaicin has been used in some patients. PMID- 11871679 TI - Information from your family doctor. Burning mouth syndrome. PMID- 11871681 TI - Information from your family doctor. Migraine headache in children and adolescents. PMID- 11871680 TI - Headaches in children and adolescents. AB - Headaches are common during childhood and become more common and increase in frequency during adolescence. The rational, cost-effective evaluation of children with headache begins with a careful history. The first step is to identify the temporal pattern of the headache--acute, acute-recurrent, chronic-progressive, chronic-nonprogressive, or mixed. The next step is a physical and neurologic examination focusing on the optic disc, eye movements, motor asymmetry, coordination, and reflexes. Neuroimaging is not routinely warranted in the evaluation of childhood headache and should be reserved for use in children with chronic-progressive patterns or abnormalities on neurologic examination. Once the headache diagnosis is established, management must be based on the frequency and severity of headache and the impact on the patient's lifestyle. Treatment of childhood migraine includes the intermittent use of oral analgesics and antiemetics and, occasionally, daily prophylactic agents. Often, the most important therapeutic intervention is confident reassurance about the absence of serious underlying neurologic disease. PMID- 11871682 TI - Evaluating and treating unintentional weight loss in the elderly. AB - Elderly patients with unintentional weight loss are at higher risk for infection, depression and death. The leading causes of involuntary weight loss are depression (especially in residents of long-term care facilities), cancer (lung and gastrointestinal malignancies), cardiac disorders and benign gastrointestinal diseases. Medications that may cause nausea and vomiting, dysphagia, dysgeusia and anorexia have been implicated. Polypharmacy can cause unintended weight loss, as can psychotropic medication reduction (i.e., by unmasking problems such as anxiety). A specific cause is not identified in approximately one quarter of elderly patients with unintentional weight loss. A reasonable work-up includes tests dictated by the history and physical examination, a fecal occult blood test, a complete blood count, a chemistry panel, an ultrasensitive thyroid stimulating hormone test and a urinalysis. Upper gastrointestinal studies have a reasonably high yield in selected patients. Management is directed at treating underlying causes and providing nutritional support. Consideration should be given to the patient's environment and interest in and ability to eat food, the amelioration of symptoms and the provision of adequate nutrition. The U.S. Food and Drug Administration has labeled no appetite stimulants for the treatment of weight loss in the elderly. PMID- 11871683 TI - Trigger points: diagnosis and management. AB - Trigger points are discrete, focal, hyperirritable spots located in a taut band of skeletal muscle. They produce pain locally and in a referred pattern and often accompany chronic musculoskeletal disorders. Acute trauma or repetitive microtrauma may lead to the development of stress on muscle fibers and the formation of trigger points. Patients may have regional, persistent pain resulting in a decreased range of motion in the affected muscles. These include muscles used to maintain body posture, such as those in the neck, shoulders, and pelvic girdle. Trigger points may also manifest as tension headache, tinnitus, temporomandibular joint pain, decreased range of motion in the legs, and low back pain. Palpation of a hypersensitive bundle or nodule of muscle fiber of harder than normal consistency is the physical finding typically associated with a trigger point. Palpation of the trigger point will elicit pain directly over the affected area and/or cause radiation of pain toward a zone of reference and a local twitch response. Various modalities, such as the Spray and Stretch technique, ultrasonography, manipulative therapy and injection, are used to inactivate trigger points. Trigger-point injection has been shown to be one of the most effective treatment modalities to inactivate trigger points and provide prompt relief of symptoms. PMID- 11871684 TI - Linezolid: its role in the treatment of gram-positive, drug-resistant bacterial infections. AB - While the choices available for the management of gram-positive, drug-resistant bacterial infections are becoming limited, antimicrobial resistance is becoming increasingly problematic because of the widespread overuse of antibiotics. Linezolid is a synthetic antibiotic belonging to a new class of antimicrobials called the oxazolidinones. Linezolid disrupts bacterial growth by inhibiting the initiation process of protein synthesis--a mechanism of action that is unique to this class of drugs. It is well absorbed with high bioavailability that allows conversion to oral therapy as soon as the patient is clinically stable. It has been approved for certain gram-positive infections including certain drug resistant enterococcus, staphylococcus, and pneumococcus strains. It is generally well tolerated, with myelosuppression being the most serious adverse effect. As a nonselective inhibitor of monoamine oxidase, caution is recommended when used with adrenergic or serotonergic agents (e.g., tyramine, dopamine, pseudoephedrine, and selective serotonin reuptake inhibitors). Judicious use of this medication should help physicians treat patients with multidrug-resistant infections. PMID- 11871685 TI - Screening for chlamydial infection: recommendations and rationale. PMID- 11871686 TI - AAP guideline on treatment of children with ADHD. American Academy of Pediatrics. PMID- 11871687 TI - The role of GC-MS and LC-MS in the discovery of drinking water disinfection by products. AB - Gas chromatography-mass spectrometry (GC-MS) has played a pivotal role in the discovery of disinfection by-products (DBPs) in drinking water. DBPs are formed when disinfectants, such as chlorine, ozone, chlorine dioxide or chloramine, react with natural organic matter in the water. The first DBP known--chloroform- was identified by Rook in 1974 using GC-MS. Soon thereafter, chloroform and other trihalomethanes were found to be ubiquitous in chlorinated drinking water. In 1976, the National Cancer Institute published results linking chloroform to cancer in laboratory animals, and an important public health issue was born. Mass spectrometry and, specifically, GC-MS became the key tool used for measuring these DBPs in water and for discovering other DBPs that were formed. Over the last 25 years, hundreds of DBPs have been identified, mostly through the use of GC-MS, which has spawned additional health effects studies and regulations. Early on, GC with low resolution electron ionization (EI)-MS was used, together with confirmation with chemical standards, for identification work. Later, researchers utilized chemical ionization (CI)-MS to provide molecular weight information and high resolution El-MS to aid in the determination of empirical formulae for the molecular ions and fragments. More recently, liquid chromatography-mass spectrometry (LC-MS) with either electrospray ionization (ESI) or atmospheric pressure chemical ionization (APCI) has been used to try to uncover highly polar DBPs that most experts believe have been missed by earlier GC-MS studies. Despite 25 years of research in the identification of new DBPs, new ones are being discovered every year, even for chlorine which has been the most extensively studied. PMID- 11871688 TI - Advances in the determination of inorganic ions in potable waters by ion chromatography. AB - Ion chromatography (IC) is now a well-established methodology for the analysis of ionic species. The technique is applicable to the determination of a wide range of solutes in many sample types, although the determination of inorganic ions in potable waters continues to be the most widely used application of ion chromatography. Many standardization and regulatory bodies, such as the American Society for Testing and Materials (ASTM), International Organization for Standardization (ISO), and US Environmental Protection Agency (EPA), have approved methods of analysis based upon IC, most of which have been published within the last decade. Recent developments in the field of IC, such as the use of higher capacity columns, larger loop injections, more complex sample preparation and detection schemes, have been incorporated into these new approved methods. These advances allow the determination of environmentally significant contaminants, such as common inorganic anions, bromate, perchlorate and chromate, at trace levels in potable waters using ion chromatography. PMID- 11871689 TI - Examples of the role of analytical chemistry in environmental risk management research. AB - Analytical chemistry is an important tier of environmental protection and has been traditionally linked to compliance and/or exposure monitoring activities for environmental contaminants. The adoption of the risk management paradigm has led to special challenges for analytical chemistry applied to environmental risk analysis. Namely, methods developed for regulated contaminants may not be appropriate and/or applicable to risk management scenarios. This paper contains examples of analytical chemistry applied to risk management challenges broken down by the analytical approach and analyte for some selected work in our laboratory. Specific techniques discussed include stable association complex electrospray mass spectrometry (cESI-MS), gas chromatography-mass spectrometry (GC-MS), split-flow thin cell (SPLITT) fractionation and matrix-assisted laser desorption time of flight mass spectrometry (MALDI-ToF-MS). Specific analytes include haloacetic acids (HAA9), perchlorate, bromate, triazine degradation products, metal-contaminated colloids and Cryptosporidium parvum oocysts. PMID- 11871690 TI - Normalization procedures for sediment contaminants in spatial and temporal trend monitoring. AB - Rational pollution, or the effectiveness of natural attenuation assessments based upon estimating the degree of contamination, critically depends on the basis of a sound normalization to take into account heterogeneous sedimentary environments. By normalizing the measured contaminant concentration patterns for the sediment characteristics, the inherent variability can be reduced and so allow a more meaningful assessment of both the spatial distributions and the temporal trends. A brief overview and guidance in the methodology available for choosing an appropriate site-specific normalization approach is presented. This is followed by general recommendations with respect to the choice of normalizer and the necessary geochemical and statistical quality assurance methods, with support from the results of recent international intercomparison exercises within the QUASH (Quality Assurance of Sample Handling) programme, as well as discussions within the International Commission on the Exploration of the Sea (ICES) working groups. The most important of these recommendations is the use of a two-tiered normalization approach including wet sieving (<63 microm), followed by an additional geochemical co-factor normalization. PMID- 11871691 TI - Interlaboratory comparability of copper complexation capacity determination in natural waters. AB - The results of a test of interlaboratory comparability for the determination of copper complexation capacity and copper-ligand complex formation constant are presented. Eight water samples comprising, six natural waters, a synthetic ligand solution and a blank solution were analysed by seven laboratories using their own methods of complexation titration. Given the wide variation that might have been possible, relatively good comparability was demonstrated amongst the variety of methods for determining copper complexation capacity. The complexation capacity data largely satisfied the predefined criterion of agreement to within 50%. This provides support for the use of metal speciation criteria in the regulation of copper in the environment. Data for the determination of complex formation constants were of poorer comparability, ranging between 10(7) and 10(12) for the same water. PMID- 11871692 TI - Multi-objective, decision-based assessment of a water quality monitoring network in a river system. AB - Water quality monitoring network design has historically tended to use experience, intuition and subjective judgement in locating monitoring stations. Better design procedures to optimize monitoring systems need to simultaneously identify significant planning objectives and consider a number of social, economic and environmental constraints. The consideration of multiple objectives may require further decision analysis to determine the preference weights associated with the objectives to aid in the decision-making process. This may require the application of an optimization study to extract such information from decision makers or experts and to evaluate the overall effectiveness of locating strategies. This paper assesses the optimal expansion and relocation strategies of a water quality monitoring network using a two-stage analysis. The first stage focuses on the information retrieval of preference weights with respect to the designated planning objectives. With the aid of a pre-emptive goal programming model, data analysis is applied to obtain the essential information from the questionnaire outputs. The second stage then utilizes a weighted multi-objective optimization approach to search for the optimal locating strategies of the monitoring stations in the river basin. Practical implementation is illustrated by a case study in the Kao-Ping River Basin, south Taiwan. PMID- 11871693 TI - Nitrate occurrence in the groundwater of the Loukkos perimeter. AB - This study assesses the occurrence of nitrate in the groundwater beneath the R'mel area of the Loukkos perimeter (north-west Morocco), which covers an approximate area of 2,560 km2 and is located between the towns of Ksar el Kebir and Larache. It also borders the Atlantic Ocean. Groundwater supplies are the principal source of drinking water in this region and there is no public drinking water network in the rural area. This perimeter has a population of about 500,000 inhabitants of which the rural population represents 60%, many of whom have depended on and used the water from the aquifers for many years. The inhabitants and farmers depend on the groundwater supplies for drinking water, crop irrigation and other uses. The plain provides the ideal conditions for agriculture and the use of chemical fertilisers has been increasing. In this study, 53 water samples were collected from wells and springs. Each well or spring was sampled once or twice during 1998-2000. Nitrate concentrations ranged from extremely low up to 144 mg L(-1). Nitrate concentrations exceeded the maximum contaminant level (MCL) of 50 mg L(-1) in 12 of the 53 groundwater samples (23%), whereas 31 of the 53 samples (58%) had nitrate concentrations of less than 25 mg L(-1). PMID- 11871694 TI - Operational problems related to the preparation of the seawater soluble fraction of crude oil. AB - Owing to the importance of dissolution and weathering processes following oil spills, this work focused on the operational (quantitative) aspects related to the dissolution of petroleum-derived products, as well as the influence of solar light on both dissolution and the photoproduction of hydrogen peroxide. Four Brazilian crude oil samples were used to study the transfer process of organic compounds from the crude oil film to the aqueous phase (natural seawater) over a period of up to 45 days. Dissolved organic carbon (DOC), measured by non dispersive infrared spectroscopy followed by high temperature catalytic combustion, was used to follow the partitioning between the two phases. Aqueous DOC values increased as a function of time (up to 15 days) until equilibrium was reached at concentrations ranging from 5 to 45 mg C L(-1). The final DOC concentration as well as the rate of dissolution depends on the nature of the crude oil. When exposed to sunlight, the dissolution was enhanced by up to 67.3%, and inorganic peroxides were generated in the concentration range from 4.5 up to 8.0 micromol L(-1) after 7.3 h irradiation. These results indicate that there is a need for a standard procedure for the production of the WSF in order to generate a more reliable tool to assess the impact of oil spills on the marine environment. PMID- 11871695 TI - Phthalate and alkyl phenol concentrations in soil following applications of inorganic fertiliser or sewage sludge to pasture and potential rates of ingestion by grazing ruminants. AB - Soil concentrations of dioctyl phthalate (DOP) and the alkyl phenols, octyl phenol (OP) and nonyl phenol (NP), after repeated surface applications of sewage sludge to pastures, were investigated. Liquid sludge was applied at a rate of 2.25 tonnes dry matter (DM) per hectare to each of three treated (T) plots on three occasions during the summer and two occasions in the early spring over a period of 2.5 years. Control (C) plots were treated with inorganic fertiliser containing amounts of nitrogen equivalent to those applied to the treated plots. At between 69 and 81 days after the application of sludge, 15 separate soil samples were collected from one half of each of the plots (Experiment 1). Concentrations (microg g(-1)) of DOP were higher (P < 0.001) than those of NP, while those of OP were generally below detectable levels. Mean soil concentrations of DOP were not significantly different in T and C plots [0.233 vs. 0.155 microg g(-1); standard error of the difference (SED) = 0.046; not significant (NS)], partly because there was already a relatively large amount of DOP present. NP concentrations were, however, significantly higher in T than in C plots (0.021 vs. 0.013 microg g(-1) SED = 0.002; P < 0.05). There was no consistent change over time in the mean soil concentrations of these compounds when sampled at intervals of 3-6 months. Concentrations in soil samples collected at monthly intervals following sludge application indicated that the variation in concentrations of these endocrine-disrupting compounds (EDC) was unrelated to time since sludge application. Rates of soil ingestion, expressed as the percentage of DM intake represented by soil, were higher during the winter than the summer (5.40 vs. 1.17; SED = 0.360; P < 0.001) and estimated daily intakes of DOP and NP were up to 150 microg and 8 microg, respectively. It is concluded that the application of sewage sludge to pasture does not increase soil concentrations of phthalate (as DOP) or alkyl phenols. Thus, the risk of increased exposure to these EDC as a result of sludge application is small. However, the small effect of sludge application on soil concentrations may be largely a reflection of the relatively high concentrations of DOP already present in the soil, which may be biologically significant. PMID- 11871697 TI - A problem-based, self-directed approach to the teaching of the principles of environmental and human toxicology. PMID- 11871696 TI - Performance of poly(vinyl alcohol) gel columns on the ion chromatographic determination of perchlorate in fertilizers. AB - Interest in possible sources of perchlorate (ClO4) that could lead to environmental release has been heightened since the Environmental Protection Agency placed this anion on its Contaminant Candidate List for drinking water. Although recent investigations have suggested that fertilizers are minor contributors to environmental perchlorate contamination overall, there is still interest in screening commercial products for possible contamination and quantitating perchlorate when it is found. Ion chromatography (IC) has been used for this application owing to its speed, low detection limits, widespread availability, and moderate ruggedness relative to other techniques. However, fertilizer matrixes complicate the IC analysis relative to potable water matrixes. In this study, the performance of poly(vinyl alcohol) gel resin IC columns (100 mm and 150 mm) was evaluated for fertilizer matrixes using method EPA/600/R-01/026. The NaOH eluent included an organic salt, sodium 4 cyanophenoxide. Detection was by suppressed conductivity. A set of 55 different field samples representing 48 products and previously used by the EPA to assess occurrence of perchlorate in fertilizers (EPA/600/R-01/049) was reanalyzed on the 150 mm column. The 100 mm column was used to further investigate the positive hits. Both columns gave satisfactory performance in fertilizer matrixes, with spike recoveries (+/- 15%), assured reporting levels (0.5-225 microg g(-1) except for one at 1,000 microg g(-1)), accuracy (relative error < 30% always and most < 15%), and precision [injection-to-injection reproducibility < 3% relative standard deviation (RSD)] comparable to those reported in other studies. Performance did not vary substantially between column lengths. Lastly, the results of this investigation provided further evidence in support of the conclusions that had been reached previously by the EPA on the occurrence of perchlorate in fertilizers. PMID- 11871698 TI - Field measurement of lead in workplace air and paint chip samples by ultrasonic extraction and portable anodic stripping voltammetry. AB - On-site measurement of lead in workplace air filter samples and paint chip samples by ultrasonic extraction and anodic stripping voltammetry (UE-ASV) was evaluated in the field during renovation and remodeling activities in residences having leaded paint. Aerosol and paint samples were collected using standard techniques, and the samples were analyzed on-site for lead content by portable UE ASV. Lead in sample extracts was subsequently determined by atomic absorption (AA) spectrometry in a fixed-site laboratory. The remaining sample extracts plus undissolved material (air filters or paint particles) were then subjected to hot plate digestion in concentrated nitric acid-30% hydrogen peroxide prior to AA analysis for lead. Field UE-ASV lead data were thereby compared to UE-AA and hot plate digestion-AA results from fixed-site laboratory lead measurement. Determination of lead in air filter samples by UE-ASV (over the range of 5 microg to approximately 800 microg Pb per sample) was extremely well correlated with lead measurement by UE-AA and hot plate digestion-AA procedures. However, a significant negative bias associated with ASV measurement was observed, and this was attributed to a matrix effect. Lead measurement in paint chip samples by UE ASV (over the range of approximately 10 to approximately 550 microg Pb g(-1)) was well correlated with lead measurement by UE-AA and hot plate digestion-AA procedures. However, correlation and precision were lower for lead measurement in paint samples as compared to aerosol samples, and a negative bias was also observed. Lead measurements by UE-AA were compared to lead determinations by hot plate digestion-AA; these data were highly correlated and demonstrated no significant bias. Thus it was concluded that the ultrasonic extraction procedure performed equivalently to hot plate digestion. It was reasoned that matrix effects due to the preparation and analysis of paint chip particles resulted in greater imprecision as well as negative bias by ASV measurement. Despite significant negative bias in this sample set, UE-ASV offers promise for on-site measurement of lead in samples of interest in occupational and environmental health. PMID- 11871699 TI - The role of size exclusion chromatography-flame atomic absorption spectrometry in the treatment chemistry of potable water. AB - The percentage composition of Al13, [AlO4Al12(OH)24(H2O)12]7+, in water treatment coagulants is an important criterion in the development and use of polymeric coagulants. Polymeric coagulants are generally used in cold climates or with highly turbid waters. Size exclusion chromatography flame atomic absorption spectrometry (SEC-FAAS) can separate Al13 and monomeric Al within 6 min. The percentage composition of Al13 and monomeric Al is determined by solving two simultaneous equations. Due to overlapping peaks, a 10% error is associated with this method of quantification. This method can be used on coagulants of varying "r values" (r=[OH-]/[Al3+]), or on mixtures of those coagulants and monomeric aluminium. PMID- 11871700 TI - Indoor radon/thoron levels and inhalation doses to some populations in Himachal Pradesh, India. AB - It is well established that some areas of Himachal Pradesh (H.P.) state of India situated in the environs of the Himalayan mountains are relatively rich in uranium-bearing minerals. Some earlier studies by our group have indicated high levels of radon (>200 Bq m(-3)) in the dwellings. It is in this context that an indoor radon/thoron survey has been carried out in selected villages of four districts in the state of H.P. This survey has been conducted as a part of a national, coordinated project using twin chamber dosemeter cups designed by the Environmental Assessment Division (EAD), Department of Atomic Energy, Govt. of India. The track-etch technique is used for calibration of plastic detector LR 115 type-II which are employed for recording alpha tracks due to radon/thoron and their daughters. Year long radon/thoron data have been collected for seasonal correlations of indoor radon/thoron in the dwellings. The indoor radon levels have been found to vary from a minimum value of 17.4 Bq m(-3) to a maximum value of 140.3 Bq m(-3). The indoor thoron levels vary from a minimum value of 5.2 Bq m(-3) to a maximum value of 131.9 Bq m(-3). The year average dose rate for the local population varies from 0.03 microSv h(-1) to 0.83 microSv h(-1). The annual exposure dose to inhabitants in all the dwellings lies below the upper limit of 10 mSv given in ICRP-65. PMID- 11871701 TI - Personal exposures and microenvironmental concentrations of particles and bioaerosols. AB - The aim of this study was to compare the personal exposure to particles and bioaerosols with that measured by stationary samplers in the main microenvironments, i.e., the home and the workplace. A random sample of 81 elementary school teachers was selected from the 823 teachers working for two councils in eastern Finland for the winter time measurement period. Bioaerosol and other particles were collected on filters by button samplers using personal sampling and microenvironmental measurements in homes and workplaces. The 24-hour sampling period was repeated twice for each teacher. Particle mass, absorption coefficient of the filter and the concentration of viable and total microorganisms were analyzed from each filter. In this paper, the study design, quality assurance principles and results of particle and bioaerosol exposure are described. The results show that particle mass concentrations, absorption coefficient and fungi were higher in personal exposure samples than in home and workplace samples. Furthermore, these concentrations were usually lower in the home than in the workplace. Bacterial concentrations were highest in heavily populated workplaces, while the viable fungi concentrations were lowest in workplaces. The fungi and bacteria results showed high variation, which emphasises the importance of quality assurance (duplicates and field blanks) in the microbial field measurements. Our results indicate that personal exposure measurements of bioaerosols in indoor environments are feasible and supplement the information obtained by stationary samplers. PMID- 11871702 TI - Fish consumption and plasma levels of organochlorines in a female population in Northern Norway. AB - Increased cancer incidence and mortality have been found among humans exposed to high levels of organochlorines (OCs), either accidentally or as industrial workers. In order to assess levels of OCs in Norwegian women north of the Arctic Circle and validate self-reported fish consumption as a surrogate measure of organochlorine body burden, concentrations of seven polychlorinated biphenyl (PCB) congeners [IUPAC Nos. CB-105, CB-118, CB-138 (+ CB-163), CB-153, CB-180, CB 183, CB-187], beta-hexachlorocyclohexane (beta-HCH), 2,2'-bis(p-chlorophenyl)-1,1 dichloroethylene (p,p'-DDE) and cis- and trains-chlordane (c-CD and t-CD) were examined in plasma samples of middle-aged women attending for health screening. Altogether, 47 of those invited (81%) completed a questionnaire and donated a suitable blood sample. The ability of questionnaire data to predict plasma levels of OCs was tested in linear and logistic regression analyses. Measured plasma concentrations were in the range reported for the general female population of other Western countries and the relative amounts of PCBs were similar to the circumpolar pattern. Intake of seagulls' eggs was a predictor of PCB congeners CB 138 (+CB-163) (p<0.05) and CB-153 (p<0.01). No other food category was positively associated with any compound. In contrast, duration of residence in the study municipality, body mass index (BMI) and lifetime lactation (months) were the best univariate predictors. There was an increase in beta-HCH, p,p'-DDE and most of the PCBs (p<0.05 for all) with increasing length of time a subject had lived in the municipality. BMI was a positive predictor for beta-HCH (OR=3.10, 95% CI 1.50 6.43, per 5 kgm(-2)), chlordane (OR=2.13, 95% CI 1.12-4.05, per 5 kgm(-2)) and CB 105 and CB-153 (p<0.05 for both). Lactation was negatively associated with all OCs (p<0.05), except chlordane and two of the PCB congeners. Time living in the municipality and lactation explained 34%, of the variance in concentration of total PCB in a multivariate model (p<0.001). The results indicate that regular consumption of fish (mostly lean species) from the Norwegian waters is not associated with an increased body burden of OCs (e.g., of importance to cancer development), although they confirm that lactation is the most important elimination route of these contaminants in women. PMID- 11871703 TI - Assessment of risk to public health posed by persistent organochlorine pesticide residues in milk and milk products in Mumbai, India. AB - The risk posed by the presence of organochlorine pesticides in milk and milk products was estimated for the population of Mumbai. To determine the levels of organochlorine pesticides in milk and milk products, a monitoring study was carried out in and around Mumbai City. 520 samples of milk and milk products of different brands were considered in this study. A survey was also conducted to determine the mean daily consumption of milk and milk products by different age groups and this data was used to evaluate the daily exposure to the public. Non cancer effects were evaluated by comparing the predicted exposure distributions to the published guidance values. For chemicals identified as potential human carcinogens, cancer risk was evaluated using standard methodology. The majority of the chlorinated pesticides identified in the milk and milk product samples studied were found to be at levels which do not pose unacceptable risks to the public, with the exception of alpha-HCH. The cancer risk estimated for this chemical slightly exceeds the US EPA guidance value. PMID- 11871704 TI - Role of halogen(I) cation-transfer mechanisms in water chlorination in the presence of bromide ion. AB - Bromide ion is rapidly converted to HOBr via BrCl by reaction with HOCl. The subsequent slow reactions of (HOCl, OCl-)/(HOBr, OBr-) mixtures are monitored directly by multiwavelength UV-vis absorbance methods and simultaneously by ion chromatographic measurement of ClO2-, ClO3-, and BrO3- (p[H+] 5.6-7.6). A first order loss of HOCl is observed which is catalyzed by trace concentrations of Br- and BrCl. Chlorite ion forms first and is subsequently oxidized to ClO3-. The loss of HOBr is slower and is second-order in HOBr, so that BrO3- formation takes longer than ClO3- formation. Under the conditions of this work, the relative yield of BrO3- increases with increase in pH. The decomposition of HOCI by bromide proceeds primarily by a series of halogen(I) cation-transfer reactions with subsequent halide release. The presence of HOCI increases the BrO3- yield three-fold from HOBr decay alone. PMID- 11871705 TI - The role of Raman spectroscopy in the analytical chemistry of potable water. AB - Advances in instrumentation are making Raman spectroscopy the tool of choice for an increasing number of chemical applications. For example, many recalcitrant industrial process monitoring problems have been solved in recent years with in line Raman spectrometers. Raman is attractive for these applications for many reasons, including remote non-invasive sampling, minimal sample preparation and tolerance of water. To a lesser extent, Raman spectroscopy is beginning to play a significant role in environmental analysis for the same reasons. At present, the environmental applications typically apply only to the most contaminated situations due to the still relatively high limits of detection. However, some emerging sampling technologies hold out the promise that Raman may soon be more widely applicable to the analytical chemistry of potable water. Herein we discuss these recent advances, summarize some examples of environmental applications to aqueous systems and suggest avenues of future developments that we expect to be most useful for potable water analysis. Also, a simplified, but detailed, theory of normal Raman scattering is presented. While resonance-enhanced Raman spectroscopy, surface-enhanced Raman spectroscopy and non-linear Raman techniques are briefly discussed, their theories and instrumental configurations are not addressed. Also, this article deals primarily with the modern dispersive Raman experiment (as opposed to the Fourier transform Raman experiment), because it seems most suited for potable water analysis. The goal of this article is to give the environmental scientist with no specialized knowledge of the topic just enough theory and background to evaluate the utility of this rapidly developing analytical tool. PMID- 11871706 TI - Raising a glass to clean water. PMID- 11871707 TI - Total organic carbon analysis as a precursor to disinfection byproducts in potable water: oxidation technique considerations. AB - In recent years, an increasing number of regulations and methodologies have begun to utilize total organic carbon (TOC) analysis for monitoring microbial contamination and/or disinfectant byproduct (DBP) precursors. This paper highlights some analytical differences and similarities between the two widely used TOC oxidation techniques: UV persulfate and high temperature combustion (HTC). Previous papers have come to different and sometimes contradictory conclusions on this subject. However, these studies either compared instruments with significantly different flow paths or TOC systems from different eras. Unlike previous studies, this paper compares two modern TOC analyzers with nearly identical flow paths for sample recovery, detection limits, and analysis of real world samples. On average, both persulfate and HTC oxidation yielded good recoveries for 10 hard to oxidize compounds and potable water samples from 5 different locations across the USA. In general, persulfate yielded more precise results because of its lower background response relative to sample response while HTC gave slightly higher results (roughly 2% to 3%) for surface water samples. PMID- 11871708 TI - Monitoring dissolved organic carbon in surface and drinking waters. AB - The presence of natural organic matter (NOM) strongly impacts drinking water treatment, water quality, and water behavior during distribution. Dissolved organic carbon (DOC) concentrations were determined daily over a 22 month period in river water before and after conventional drinking water treatment using an on line total organic carbon (TOC) analyzer. Quantitative and qualitative variations in organic matter were related to precipitation and runoff, seasons and operating conditions. Following a rainfall event, DOC levels could increase by 3.5 fold over baseflow concentrations, while color, UV absorbance values and turbidity increased by a factor of 8, 12 and 300, respectively. Treated water DOC levels were closely related to the source water quality, with an average organic matter removal of 42% after treatment. PMID- 11871709 TI - A brief overview of modern directions in marine DOC studies part I.- Methodological aspects. AB - The necessity for determining the role of dissolved organic carbon (DOC) in the global carbon cycle stimulated the development of different methods of DOC analysis in aquatic environments. Progress in this direction has been made by oceanographers who developed and introduced a high-temperature catalytic oxidation (HTC) method for low organic carbon concentrations. Today this method is the reference method for marine DOC study. The combination of available reference materials and the participation in intercalibration exercises has resulted in both an increased accuracy and higher precision for this method. The HTC method completely oxidizes the more resistant DOC; makes information rapidly available following the completion of the field analysis; provides a high precision (down to 0.5 microM C); covers the range of seawater DOC concentrations (35-80 microM C and higher); with certain modifications it has proved to be both seaworthy and amenable to automated analysis; and the reliable and relatively easy to operate HTC analyzer is commercially available and easily combined with a total nitrogen analyzer for simultaneous measurements of both parameters in the same sample. In this review we summarize some aspects of sample collection, handling and the analytical chemistry of the DOC analysis by the HTC technique in marine study. PMID- 11871710 TI - A brief overview of modern directions in marine DOC studies part II--recent progress in marine DOC studies. AB - Progress made in analytical techniques allows the formulation of new concepts in the biogeochemistry of organic carbon. The second part of our review summarizes the latest evolution and introduces new ideas in the biogeochemistry of marine dissolved organic carbon (DOC). Via classification of different fractions and sources of DOC, characterization of its composition, age and availability for bacterial utilization, and fate of DOC, we show the role of DOC in the global carbon cycle and the significance of bulk DOC in the oceans. Special emphasis is placed on the microbial loop in the cycling of DOC and its relation with higher trophic levels (phytoplankton and zooplankton). Significant progress has also been made in the study of the roles of colloidal organic material in metal complexation, ultraviolet radiation in dissolved organic matter photochemical oxidation, and chromophore-containing constituents of DOC as the signature of DOC for satellite observations. The importance of bulk DOC in the global carbon cycle requires the inclusion of this fraction in the regional and global carbon models. We predict that future DOC study in the ocean will focus on the development of sophisticated, almost continuously recording, moored DOC instrument arrays for the monitoring of small-scale DOC horizontal and vertical patchiness; widespread time series stations including estuarine, coastal and open environments; more detailed chemical characterization of different fractions of organic carbon from diverse marine habitats; parameterization of predictive models of DOC cycling on regional and global scales, incorporating the microbial loop; and finally, monitoring of DOC dynamics from satellites on regional and global scales. PMID- 11871711 TI - Assessment of relative accuracy in the determination of organic matter concentrations in aquatic systems. AB - Accurate determinations of total (TOC), dissolved (DOC) and particulate (POC) organic carbon concentrations are critical for understanding the geochemical, environmental, and ecological roles of aquatic organic matter. Of particular significance for the drinking water industry, TOC measurements are the basis for compliance with US EPA regulations. The results of an interlaboratory comparison designed to identify problems associated with the determination of organic matter concentrations in drinking water supplies are presented. The study involved 31 laboratories and a variety of commercially available analytical instruments. All participating laboratories performed well on samples of potassium hydrogen phthalate (KHP), a compound commonly used as a standard in carbon analysis. However, problems associated with the oxidation of difficult to oxidize compounds, such as dodecylbenzene sulfonic acid and caffeine, were noted. Humic substances posed fewer problems for analysts. Particulate organic matter (POM) in the form of polystyrene beads, freeze-dried bacteria and pulverized leaf material were the most difficult for all analysts, with a wide range of performances reported. The POM results indicate that the methods surveyed in this study are inappropriate for the accurate determination of POC and TOC concentration. Finally, several analysts had difficulty in efficiently separating inorganic carbon from KHP solutions, thereby biasing DOC results. PMID- 11871712 TI - Monochloramine loss in the presence of humic acid. AB - Free chlorine has been used extensively as a primary and secondary disinfectant for potable water. Where it is difficult to maintain a free chlorine residual or when disinfection by-products (DBPs) are of concern, monochloramine has been used to provide a stable disinfectant residual in distributions systems. Reactions of disinfectants, free chlorine or monochloramine, with natural organic matter (NOM) consequently result in the formation of DBPs such as trihalomethanes and haloacetic acids. However, few studies have focused on the fate and kinetics of monochloramine loss in the presence of reactive constituents such as NOM. Monochloramine is inherently unstable and decays even without reactive constituents present via a mechanism known as autodecomposition. Therefore, to predict monochloramine concentrations in the presence of NOM is clearly associated with the ability to adequately model autodecomposition. This study presents the results of a semi-mechanisiic model capable of predicting the loss of monochloramine in the presence of humic material in the pH range of 6.55-8.33. The model accounts for both fast and a slow monochloramine demand to explain the loss of monochloramine over the pH range of this study. The formation of dichloroacetic acid was also predicted due to the ability of the model to differentiate monochloramine reaction pathways in the presence NOM. The results shown here demonstrate the ability of a semi-mechanistic model to predict monochloramine residuals and DBP formation in the presence of humic material. PMID- 11871713 TI - Method for the determination of sub-ppm concentrations of perfluoroalkylsulfonate anions in water. AB - The determination of sub-ppm concentrations of aqueous perfluoroalkylsulfonate (PFSt) anions, including perfluorooctylsulfonate (PFOS), has been accomplished with a relatively simple mass spectrometric procedure that does not require extraction of the analytes into an organic solvent or a chromatographic separation prior to injection into the negative-ion electrospray ionization mass spectrometer. Sample pretreatment was minimized and consisted of dilution of the aqueous samples of groundwater, surface water, tap water, and distilled water with acetonitrile, addition of dodecylsulfate (DDS) as an internal standard, and, in some cases, addition of known amounts of perfluorobutylsulfonate (PFBS) or PFOS for standard-addition experiments. The linear-response range for PFOS is 25.0 microg L(-1) to 2.5 mg L(-1). The lower limit of this range is three orders of magnitude lower than an equally straightforward chromatographic method. The relative errors for standard aqueous solutions containing only 25.0 microg L(-1) and 2.5 mg L(-1) PFOS are +/- 14% and +/- 7%, respectively, with 133 microg L(-1) DDS as the internal standard. The detection limit and quantification limit for PFOS in these standards are 5.0 microg L(-1) and 25.0 microg L(-1), respectively. Six different PFS anions, containing three to eight carbon atoms, were identified and quantified in an aqueous film-forming foam (AFFF) formulation using the method of standard additions. Two alkylsulfate anions and two perfluoroalkylcarboxylate anions were also identified in the AFFF formulation. PMID- 11871714 TI - Biological reduction of perchlorate in ion exchange regenerant solutions containing high salinity and ammonium levels. AB - The most promising technologies to remove perchlorate from water are ion exchange and biological reduction. Although successful, ion exchange only separates perchlorate from water; it does not eliminate it from the environment. The waste streams from these systems contain the caustic or saline regenerant solutions used in the process as well as high levels of perchlorate. Biological reduction could be used to treat the regenerant waste solutions from the ion exchange process. A treatment scheme, combining ion exchange and biodegradation, is proposed to completely remove perchlorate from the environment. Perchlorate-laden resins generate brines containing salt concentrations up to 6% or caustic solutions containing up to 0.5% ammonium. Both, high salt and ammonium hydroxide concentrations are potentially toxic to microorganisms. Therefore, the challenge of the proposed system is to find perchlorate reducing microorganisms that are effective under such stressful conditions. Preliminary results have shown that salt concentrations as low as 0.5% reduced the perchlorate biodegradation rate by 30%; salt concentrations greater than 1% decreased this rate to 40%. Although biodegradation was seen in ammonium levels of 0.4%, 0.6% and 1%, the perchlorate biodegradation rate was 90% of that at 0% ammonium hydroxide. Further research will focus on the isolation and/or acclimation of microorganisms that are able to biodegrade perchlorate under these stressful conditions. PMID- 11871715 TI - Homosexuality in French history and culture. Introduction. PMID- 11871716 TI - "Au sein de vos pareilles": sapphic separatism in late eighteenth-century France. PMID- 11871717 TI - The Palais-Royal and the homosexual subculture of nineteenth-century Paris. PMID- 11871718 TI - Les Chevaliers de la guirlande: cellmates in restoration France. PMID- 11871719 TI - Homosexuals in the city: representations of lesbian and gay space in nineteenth century Paris. PMID- 11871720 TI - Pederasts, prostitutes, and pickpockets in Paris of the 1870s. PMID- 11871721 TI - Drames d'amour des pederastes: male same-sex sexuality in Belle Epoque print culture. PMID- 11871722 TI - Homosexuality in the French colonies. PMID- 11871724 TI - Female friendship as the foundation of love in Madeleine de Scudery's "Histoire de Sapho". PMID- 11871723 TI - Folles, swells, effeminates, and homophiles in Saint-Germain-des-Pres of the 1950s: a new "precious" society? PMID- 11871725 TI - The birth of a French homosexual press in the 1950s. PMID- 11871726 TI - The construction of a political and media presence: the homosexual liberation groups in France between 1975 and 1978. PMID- 11871727 TI - Gay mimesis and misogyny: two aspects of the same refusal of the other? PMID- 11871728 TI - Masculinity and satires of "sodomites" in France, 1660-1715. PMID- 11871729 TI - "That friendship which possesses the soul": Montaigne loves La Boetie. PMID- 11871731 TI - The "Italian taste" in the time of Louis XVI, 1774-92. PMID- 11871730 TI - The abominable Madame de Murat. PMID- 11871732 TI - "Brutal passion" and "depraved taste": the case of Jacques-Francois Pascal. PMID- 11871733 TI - Local antibiotic lock for the treatment of infections related to central catheters in parenteral nutrition in children. AB - BACKGROUND: Central venous catheter-related (CVC) infections represent the most common complication of parenteral nutrition. These infections are usually treated by means of long-term systemic antibiotic treatment. The objective of this study was to determine the efficacy of combining a local antibiotic lock with a short systemic double antibiotic to treat CVC-related staphylococci infections. METHODS: Any child with coagulase-negative staphylococci or Staphylococcus aureus septicemia, confirmed by a positive blood culture, was included in the study. A double antibiotic systemic treatment composed of amikacin and teicoplanin was started and continued for 5 days. The antibiotic treatment was combined from the first day (D0) with a local teicoplanin lock, which was left for 12 hours a day in the catheter for 15 days. Parenteral nutrition was continued on a nocturnal cyclic mode during antibiotic treatment. The efficacy of the treatment was evaluated by clinical (body temperature), biologic [C-reactive protein levels (CRP)], and bacteriologic (blood culture) measures. RESULTS: Twenty CVC-related infection episodes in 13 patients were analyzed for the study. In the initial biologic test, CRP varied from 2 to 130 mg/L (mean 43 mg/L). After 3 days of treatment, CRP varied from 2 to 61 mg/L (mean 12 mg/L). The median time until normalization of temperature and CRP levels after the beginning of antibiotic treatment was 3.2 days (range 1 to 14 days) and 6.2 days (range 2 to 19 days), respectively. All blood cultures were negative for infection 48 hours after stopping the treatment. Only 1 therapeutic failure was observed during the treatment. The patient had persistent signs of clinical septicemia that required removal of the CVC. Two catheter-related infection recurrences were observed in the month after termination of the local antibiotic lock, which also required removal of the CVC. The central venous catheter was maintained in the other cases. CONCLUSIONS: Teicoplanin antibiotic locks, combined with a short conventional systemic antibiotic treatment and continuation of cyclic parenteral nutrition, seem effective and well-tolerated treatments for CVC infections. PMID- 11871734 TI - Efficacy of antibiotic-lock technique with teicoplanin in staphylococcus epidermidis catheter-related sepsis during long-term parenteral nutrition. AB - BACKGROUND: The antibiotic-lock technique has been suggested to treat catheter related sepsis in parenteral nutrition and to avoid catheter removal. METHODS: To determine the incidence of catheter-related sepsis, the bacteria involved, and the efficacy of the antibiotic-lock technique with teicoplanin, all patients (n = 263) undergoing parenteral nutrition from January 1997 to December 1999 in one center, with patients at the hospital (n = 209) and at home (54) were retrospectively studied. The antibiotic-lock technique with teicoplanin was systematically used in all suspected infections and maintained in staphylococcus epidermidis (SE) infections. RESULTS: A total of 21 of 263 patients had 34 infections (0.11/patient per year): 12 of 209 hospitalized and 9 of 54 home patients. A total of 10 of 34 infections were due to non-SE, and the catheter was immediately removed. The other 24 of 34 infections were due to SE; in 5 of 24, the catheter was removed after 48 hours of the antibiotic-lock technique because of persistent fever or thrombosis. A total of 5 of 12 patients had 2 or more infections on the same catheter. The antibiotic-lock technique prevented short term catheter removal in these cases, but a second infection occurred within a median of 50 days. In 4 of 5 cases, a third infection occurred in a mean delay of 90 days so that the catheter was removed. In 3 of 5 patients, bacteria was analyzed with pulsed field gel electrophoresis, which showed that recurrent infections were due to the same strain in all cases. CONCLUSIONS: In this study, the incidence of catheter-related sepsis was low and mostly related to SE. Our results do not support the use of the teicoplanin antibiotic-lock technique in SE infections. PMID- 11871736 TI - With bioimpedance spectroscopy, the errors get fat when the patients get slim. PMID- 11871735 TI - Iron deficiency anemia in patients receiving home total parenteral nutrition. AB - BACKGROUND: Certain patients receiving home total parenteral nutrition (HPN) are likely to develop iron-deficiency anemia because of inadequate absorption or chronic iron loss from gastrointestinal lesions. The objective of this study was to examine the incidence and prevalence of iron deficiency anemia in patients on long-term HPN (>6 months) and to investigate both the efficacy of and rate of adverse reactions to parenteral iron dextran therapy. METHODS: The records of 55 patients treated with HPN for >6 months between January 1, 1994 and December 31, 1999 were examined. RESULTS: Thirty patients (55%) had evidence of iron deficiency anemia. Ten patients were diagnosed at the initiation of HPN, and in 20 patients, iron deficiency developed after receiving HPN. The time between initiation of HPN and development of anemia ranged from 2 to 97 months (mean 28.8+/-26.2 months). Mild iron loss from the gastrointestinal tract seemed to be the predominant reason for iron deficiency. Regular treatment with small amounts of iron in HPN appeared to be safe and efficacious, with no reported side effects. Total dose infusion of iron was associated with adverse reactions in as many as 25% of these patients, although all reactions were mild and self-limited. CONCLUSIONS: Iron-deficiency anemia is common in patients receiving chronic HPN. Regular small doses of iron in HPN formula, rather than total dose infusion, is the preferred treatment. PMID- 11871737 TI - Influence of glycine on intestinal ischemia-reperfusion injury. AB - BACKGROUND: It has been reported that glycine may protect donor small intestine against hypothermic ischemia before transplantation. This is consistent with the documented role of glycine as a natural cytoprotectant. OBJECTIVE: Using an in vivo rodent model, we sought to determine whether exposure to a 20% glycine solution reduces the extent of warm ischemia-reperfusion injury. METHODS: Wistar rats (n = 50) underwent laparotomy. A baseline group did not receive any further intervention. The remaining animals had cannulation of the aorta before the initiation of intestinal ischemia (30 minutes) followed by reperfusion (30 minutes). Using a factorial design, rats were randomized to receive local tissue perfusion with either normal saline or a 20% glycine solution during either the preischemia or the prereperfusion phase. Standardized segments of small intestine were removed at the end of the study period to determine the extent of ischemia reperfusion injury. RESULTS: Perfusion with 20% glycine increased mucosal protein content (p < .05), increased mucosal DNA content (p < .05), reduced intestinal myeloperoxidase activity (p < .05), and maintained mucosal glutaminase activity. This was true regardless of whether glycine was administered during the preischemia phase or the prereperfusion phase. CONCLUSIONS: Local perfusion with 20% glycine can diminish warm ischemia-reperfusion injury to the rat small intestine in an in vivo model. The role of glycine supplementation should be evaluated in situations where hemodynamic instability may be responsible for breakdown in the gut barrier. PMID- 11871738 TI - Qualitative manipulation of amino acid supply during total parenteral nutrition in surgical patients. AB - We investigated whether a qualitative manipulation of amino acid mixture could improve the nutritional status of patients undergoing surgery. Patients received total parenteral nutrition for 2 consecutive 5-day periods. Energy and nitrogen supplies were calculated using a modified Harris-Benedict equation, with an energy-to-nitrogen ratio of 125 kcal/g nitrogen (N). The mean kilocalorie lipids to-glucose ratio was 35% to 65%. There were 6 patients in the control group and 7 patients in the experimental group. The control group received the same standard amino acid solution (Vintene, Baxter-Clintec, Maurepas, France) for the entire 10 days. The experimental group received the standard solution during the first 5 days but was switched to a more individualized solution during the last 5 days. The second solution was determined from a linear regression performed on day 3 comparing rate of infusion of each amino acid and its plasma variations after 3 hours of infusion with basal values. Amino acids were defined as oversupplied or undersupplied when not within the 95% confidence interval (above or below the curve, respectively). Daily nitrogen balance and urinary excretion of 3 methylhistidine were measured from day 1 to day 10 in all patients. For the patients in whom these measures were made, amino acid variations except those of lysine (11 of 12), ornithine (6 of 12), alanine (5 of 12), arginine (5 of 12), and glutamate (5 of 12) were within the 95% confidence interval. During the second 5-day period, imbalances persisted in the control group but were almost gone in the experimental group. Daily nitrogen balance was not significantly different between groups. However, when expressed as a mean over each period, nitrogen balance was significantly higher during the second period in the experimental group than in the control group: 4.5+/-0.8 g N/d versus 0.2+/-0.7 g N/d, p < .01. The ratio of urinary 3-methylhistidine to creatinine decreased from day 1 to day 10 in both groups. These findings suggest that the relationship between rate of infusion and plasma amino acid variation may offer a rational basis for choosing the most appropriate amino acid mixture for catabolic patients. An appropriate mixture would limit plasma amino acid imbalances and improve nitrogen retention. This therapy must now be tested in a larger population of patients. PMID- 11871739 TI - 2001 Harry M. Vars Research Award. Enteral nutrients alter enterocyte function within an in vitro model similar to an acute in vivo rat model during hypoxia. AB - BACKGROUND: Early enteral nutrition in patients following traumatic injury is an important intervention. However, after shock-resuscitation, intestinal hypoperfusion persists despite adequate systemic resuscitation. Our previous in vivo rat studies indicate that hypoperfusion impairs mucosal function in the small intestine. Therefore, the current study sought to improve previous in vitro models by the following means: (1) We used Caco-2 monolayers stably transfected with the brush-border sodium-glucose co-transporter (SGLT-1); and (2) we created an environment that mimicked the physiologic enterocyte environment. We hypothesized that hypoxic alterations of epithelial function in an in vitro model are comparable to those of an in vivo rat model. METHODS: After 21 days, monolayers were randomized to receive 24 hours of incubation in a normoxic or hypoxic environment. Cells were further randomized to receive 1 of 4 nutrient treatments: mannitol (an osmotic control), glucose (uses SGLT-1 and is metabolized), 3-O-methylglucose (3-O-mg; uses SGLT-1 and is not metabolized), or fructose (does not use SGLT-1 but can be metabolized). RESULTS: Transepithelial resistance (p = .007) and short-circuit current (p = .05) were lower in hypoxic groups. When compared with normoxic groups, hypoxic groups had significantly impaired glucose (p < .001) but not glutamine transport, irrespective of nutrient treatment. Additionally, adenosine triphosphate/adenosine diphosphate ratio was reduced (p = .01) and lactate concentration was increased (p < .001) during hypoxia. CONCLUSIONS: In summary, results from this in vitro study using Caco 2BBe cells stably transfected with SGLT-1 correspond to results obtained in the in vivo rat model. Therefore, this is an appropriate in vitro model in which to study cellular alterations caused by the hypoxic small intestine, with the goal of ensuring safe early enteral nutrition following traumatic injury. PMID- 11871740 TI - Magnesium, calcium, zinc, and nitrogen loss in trauma patients during continuous renal replacement therapy. AB - BACKGROUND: Whether standard nutrition support is sufficient to compensate for mineral loss during continuous renal replacement therapy (CRRT) is not known. METHODS: Adult men with traumatic injuries were recruited; one-half of recruits required CRRT for acute renal failure. All urine and effluent (from CRRT) were collected for 72 hours. Urine, effluent, and dialysate were analyzed for magnesium, calcium, and zinc using atomic absorption spectrometry. Urea nitrogen in blood, urine, and effluent were determined by measuring conductivity changes after the addition of urease. Blood was analyzed for magnesium and calcium as part of routine care. Intake was calculated from orders and intake records. RESULTS: Patients receiving CRRT (n = 6) lost 23.9+/-3.1 mmol/d (mean +/- SEM) of magnesium and 69.8+/-2.7 mmol/d of calcium compared with 10.2+/-1.2 mmol/d and 2.9+/-2.5 mmol/d, respectively, lost in patients not in acute renal failure (n = 6; p < .01). Zinc intake was significantly greater than loss in both groups (p < .03). Urea nitrogen excretion did not differ between groups. Serum magnesium was 0.75+/-0.04 mmol/L for CRRT patients, significantly lower than the 0.90+/-0.03 mmol/L for control patients (p < .01). Total blood calcium was below normal in both groups; ionized calcium was below normal in CRRT patients. CONCLUSIONS: CRRT caused significant loss of magnesium and calcium, necessitating administration of more magnesium and calcium than was provided in standard parenteral nutrition formulas. However, additional zinc was not required. CRRT removed amounts of urea nitrogen similar to amounts removed by normally functioning kidneys. PMID- 11871741 TI - Homocysteine blood level in long-term care residents with oropharyngeal dysphagia: comparison of hand-oral and tube-enteral-fed patients. AB - BACKGROUND: Levels of homocysteine, methylmalonic acid, and relevant vitamins were measured and evaluated in patients with oral dysphagia (OD) receiving long term care (LTC). METHODS: Group A was composed of 26 orally fed patients, and group B was composed of 25 patients who were fed by nasogastric tube. All patients were hospitalized in the LTC departments of the Geriatric Medical Center, Shmuel Harofe. General and nutritional status were assessed, and levels of vitamins including B12, folate, and B6 were measured along with serum homocysteine and urine methylmalonic acid levels. RESULTS: Homocysteine levels were significantly higher in the orally fed patients (p < .001); 92% had higher than-normal homocysteine levels. The levels of vitamins B12, folate, and B6 were significantly lower in the orally fed patients (p < .001), although in most patients levels were in the normal range. Notably, the levels of homocysteine were significantly correlated with the levels of all vitamins but not with the albumin or hemoglobin values. Levels of methylmalonic acid were also higher in the orally fed patients, but the difference was not statistically significant, and there was no correlation between vitamin level and methylmalonic acid level. CONCLUSIONS: Orally fed patients with OD have substantially higher homocysteine levels and appreciably lower levels of the relevant vitamins than patients with OD who receive tube feeding. Therefore, homocysteine measurement may be the preferred indicator of vitamin intake in orally fed patients with OD. PMID- 11871742 TI - Short-term efficacy of enteral nutrition in the treatment of active Crohn's disease: a randomized, controlled trial comparing nutrient formulas. AB - BACKGROUND: The optimal dietary fat content to induce clinical remission in active Crohn's disease has been the subject of controversy. We therefore performed a prospective, randomized, controlled study to compare the effects of nutrient formulas differing in the amount of medium-chain triglycerides (MCT). METHODS: Thirty-six patients with active Crohn's disease whose Crohn's disease activity index (CDAI) was > or =150 were included in the study. A formula with 3.4 g of fat per 2000-kcal dose was used as the nutrient formula with a low-fat content (ED group), and a formula with 55.6 g of fat per 2000-kcal dose was used as the nutrient formula with a high amount of MCT (TL group). RESULTS: The rate of short-term remission induction at 6 weeks was 67% in the ED group and 72% in the TL group (p = NS). Therapy markedly reduced the high CDAI and van Hees activity index in both groups, with no significant difference in the pattern of the time-course changes. C-reactive protein levels, erythrocyte sedimentation rate, and low serum albumin and plasma prealbumin levels normalized over the course of therapy, with no significant difference between the 2 groups. The assessment of fatty acid fractions revealed that the triene/tetraene ratio began to increase at 2 weeks in the ED group. The serum levels of linoleic acid, an omega-6 fatty acid, almost always varied within the normal range during the treatment period in the TL group, but in the ED group, levels began to decrease significantly at 2 weeks. The levels of linolenic acid, an omega-3 fatty acid, decreased in both groups. CONCLUSIONS: Both nutrient formulas induced clinical remission in about two-thirds of patients. The results of the present study suggest that it is not necessary to restrict the amount of MCT when given in liquid form to patients with active Crohn's disease. PMID- 11871743 TI - Rapid Bielschowsky silver impregnation method using microwave heating. AB - The Bielschowsky silver impregnation method has been used extensively to demonstrate neuronal processes including dendrites, axons and neurofibrils. In this study, we examined the differences in the time required for and the staining quality of the Bielschowsky method for neuronal processes when microwave heating was used instead of processing at room temperature. For this purpose, a control group of sections stained according to the conventional method at room temperature was compared to an experimental group stained in a microwave oven at 180 W for 2, 4 and 1 min in 2% silver nitrate, ammoniacal silver nitrate and gold chloride, respectively. Light microscopic examination demonstrated that the normal structure was preserved in both groups and that there was no difference in the staining quality between the control and the microwave groups. In addition, staining time for this procedure was reduced to 8 min by using the microwave oven. Our study revealed that microwave irradiation can be used safely for Bielschowsky silver impregnation of neuronal tissues. PMID- 11871744 TI - Phosphate ions in root-tip dividing cells: a combined trapping and squash method with implications for nuclear transcription. AB - We examined the pattern of inorganic orthophosphate (PPi) ion distribution in dividing cells of Zea mays root-tips. Unfixed and paraformaldehyde- or glutaraldehyde-vapor fixed tissues were immersed in lead acetate, glutaraldehyde, and cacodylate buffer to capture PPi as insoluble orthophosphate lead hydroxyapatite. Excess lead ions were removed with sodium citrate, then permeabilized in ammonia. Precipitates were stained with potassium sulfide, washed with distilled water and squashed in a drop of glycerin. The accumulation of PPi ions was cyclic in the cytoplasm during mitosis and they surrounded all chromosomes during metaphase and anaphase. Partition between dividing cells started with a high concentration of PPi ions at sites where plasma membrane and cell walls formed. Small daughter cells and those in G1 phase had PPi concentrated in the nucleolus, with lower levels elsewhere in the nucleus. Later in the cell cycle, there were greater amounts of PPi ions associated with condensed chromatin in larger nuclei. In Xenopus laevis oocytes, PPi was concentrated in the nucleus, mainly in the active central core of multiple nucleoli. These results and others indicate that compartmentalization of PPi occurs in the intact cell and correlates with the rate of transcription in distinct functional domains within the nucleus. PMID- 11871745 TI - Staining of intracellular deposits of uranium in cultured murine macrophages. AB - In our studies of the health effects of internalized depleted uranium, we developed a simple and rapid light microscopic method to stain specifically intracellular uranium deposits. Using J774 cells, a mouse macrophage line, treated with uranyl nitrate and the pyridylazo dye 2-(5-bromo-2-pyridylazo)-5 diethylaminophenol, uranium uptake by the cells was followed. Specificity of the stain for uranium was accomplished by using masking agents to prevent the interaction of the stain with other metals. Prestaining wash consisting of a mixture of sodium citrate and ethylenediaminetetraacetic acid eliminated staining of metals other than uranium. The staining solution consisted of the pyridylazo dye in borate buffer along with a quaternary ammonium salt, ethylhexadecyldimethylammonium bromide, and the aforementioned sodium citrate/ethylenediaminetetraacetic acid mixture. The buffer was essential for maintaining the pH within the optimum range of 8 to 12, and the quaternary ammonium salt prevented precipitation of the dye. Staining was conducted at room temperature and was complete in 30 min. Staining intensity correlated with both uranyl nitrate concentration and incubation time. Our method provides a simple procedure for detecting intracellular uranium deposits in macrophages. PMID- 11871746 TI - A new method for surface staining large slices of fixed brain using a copper phthalocyanine dye. AB - Here we describe a method for gross staining of gray matter in slices of formaldehyde-fixed human brain. After protection of white matter with 4% phenol at 60 degrees C for 5 min followed by a cold water wash, the gray matter was stained for 10-15 min at 20-25 degrees C with 1% aqueous copper(II) phthalocyanine tetrasulfonic acid tetrasodium salt (CPTS). The staining resisted all attempts to be washed from the gray matter. Stained slices can be stored indefinitely in slightly acidified water, or plastinated as permanent dry specimens. PMID- 11871747 TI - Development of a genomic in situ hybridization method using Technovit 7100 sections of early wheat embryo. AB - It is difficult to observe the behavior of chromosomes in early wheat embryos because they are wrapped in several cell layers of the ovary. Here we conducted genomic in situ hybridization on sections of ovary embedded in Technovit 7100, a resinous compound suitable for in situ hybridization of mRNA in sectioned tissues. With this resin it is possible to make thin sections with high resolution, no autofluorescence, and good water permeability. These features enable histochemical study using fluorescence microscopy. We established the most suitable conditions for the denaturation of target DNA embedded in Technovit resin, and performed GISH on them. Using this method, we identified Leymus mollis chromosomes in the young ovary of F1 hybrids between wheat and L. mollis. Furthermore, we observed the behavior of maize chromosomes in early wheat x maize hybrid embryos. PMID- 11871749 TI - Effectiveness of local exhaust for reducing welding fume exposure during boiler rehabilitation. PMID- 11871748 TI - Classification and naming of dyes, stains and fluorochromes. AB - A classification of dyes and other colorants is proposed, based on the chemical features responsible for their visibility and generally consonant with the writings of modern color chemists. The scheme differs in several respects from that of the Colour Index (CI), but it retains some traditional small groups of dyes that include biological stains. Natural dyes, recognized as a group in the CI, are placed with or near synthetic dyes with identical or similar chromophores. The new scheme also provides categories for dyes and fluorochromes that do not have places in the CI classification. Some CI categories, including lactones, aminoketones and hydroxyketones, are not recognized in this new scheme, which is adopted in the forthcoming 10th edition of Conn's Biological Stains: a Handbook of Dyes and Fluorochromes for Use in Biology and Medicine. Some rules are also set out for the spelling of trivial names, which has long been inconsistent in scientific literature. The ending '-ine' is used for compounds derived from organic bases (e.g., fuchsine and thionine, not fuchsin or thionin), and names ending in '-in' are for compounds that are not bases or their derivatives (e.g., eosin and phloxin, not eosine or phloxine). Initial capital letters are used only for words that are names of people or places (e.g., Nile blue or Congo red) and for the 'generic' components of CI application names (as in Acid yellow 36). Other words, including trade names that have fallen into common usage are not capitalized (e.g., alcian blue, biebrich scarlet, coomassie blue). The recommended spellings of some dyes differ from those commonly seen in vendors' catalogs and in biological publications, but they are generally consistent with English and American dictionaries, with recent writings in English by color chemists, and with the trivial names of other organic compounds. PMID- 11871750 TI - Carbon monoxide poisoning and death after the use of explosives in a sewer construction project. PMID- 11871751 TI - Employee exposure to high-level radio frequency radiation. PMID- 11871752 TI - Selecting isocyanate sampling and analytical methods. PMID- 11871753 TI - Analysis of quartz by FT-IR in air samples of construction dust. AB - The construction industry is reported to have some of the highest exposures to silica-containing dust. With the designation of crystalline silica as a group I human carcinogen by the International Agency for Research on Cancer (IARC), there exists a need for an analytical method to accurately quantify low levels of quartz. A method is described that uses FT-IR for quartz analysis of personal air samples collected from heavy and highway construction sites using 4-stage personal impactors. Sample filters were ashed and 13-mm or 5-mm pellets were prepared. Absorbance spectra were collected using FT-IR at resolution of 1 cm(-1) and 64 scans per spectrum. Two spectra were collected per sample using the appropriate background spectrum subtraction. Spectral manipulations such as Fourier self-deconvolution and derivatizations were performed to improve quantification. Peak height for quartz was measured at 798 cm(-1) for quantitative analysis. The estimated limit of detection for the 5-mm pellets was 1.3 microg. Recoveries of Min-U-Sil 5 spikes showed an average of > or = 94 percent for the two pellet types. The coefficient of variation of the 5-mm pellet was 9 percent at 6 microg quartz load, and 7 percent at 62 microg load. Interferences from clay, amorphous silica, concrete, calcite, and kaolinite were investigated, these being the more likely sources of interferences in construction environment. Spikes of mixtures of amorphous silica or kaolinite with Min-U-Sil 5 showed both contaminants introduced, on average, a positive error of < 5 microg with average recoveries of 106 percent and 111 percent, respectively. Spikes of mixtures of clay or concrete with Min-U-Sil 5 showed overall average recovery of 100 percent and 90 percent, respectively, after accounting for the presence of quartz in clay and concrete. This method can quantify low levels of quartz with reasonable accuracy in the face of common contaminants found in the construction industry. PMID- 11871754 TI - Limiting metabolic rate (thermal work limit) as an index of thermal stress. AB - The development of a rational heat stress index called thermal work limit (TWL) is presented. TWL is defined as the limiting (or maximum) sustainable metabolic rate that euhydrated, acclimatized individuals can maintain in a specific thermal environment, within a safe deep body core temperature (< 38.20 degrees C) and sweat rate (< 1.2 kg/hr(-1)). The index has been developed using published experimental studies of human heat transfer, and established heat and moisture transfer equations through clothing. Clothing parameters can be varied and the protocol can be extended to unacclimatized workers. The index is designed specifically for self-paced workers and does not rely on estimation of actual metabolic rates, a process that is difficult and subject to considerable error. The index has been introduced into several large industrial operations located well inside the tropics, resulting in a substantial and sustained fall in the incidence of heat illness. Guidelines for TWL are proposed along with recommended interventions. TWL has application to professionals from both the human and engineering sciences, as it allows not only thermal strain to be evaluated,. but also the productivity decrement due to heat (seen as a reduced sustainable metabolic rate) and the impact of various strategies such as improved local ventilation or refrigeration to be quantitatively assessed. PMID- 11871755 TI - Assignment of skin notation for maximum allowable concentration (MAC) list in Poland. AB - Organic chemicals from the Polish maximum allowable concentration (MAC) list were analyzed for skin notation. It can be concluded that the dermal dose LD50s determined on experimental animals ought to be adopted as the fundamental criterion for providing a substance with the percutaneous absorption notation in the MAC list. All chemicals with LD50s. value below 1,000 mg/kg should be provided with the Sk index in the MAC list. For other chemicals, a skin notation would be considered when repeated human and dermal application tests have shown significant systemic effects following exposure. When information on the characteristics specified above were not available, physicochemical data required to calculate the flow (solubility, octanol/water partition coefficient, molecular weight) were obtained to consider a skin notation. PMID- 11871756 TI - Field comparison of 37-mm closed-face cassettes and IOM samplers. AB - On examining the published results of comparisons of sampling with Institute of Occupational Medicine (IOM) (Edinburgh, U.K.) samplers and 37-mm closed-face cassettes it was observed that they usually do not take into account the dust deposited on the walls of the cassettes. As the method used by the Institut National de Recherche et de Securite, France (INRS), to analyze metals includes the analysis of these deposits, it was decided to evaluate the differences obtained between these samplers when using this method. The essays were conducted in three different plants, and repetitive static samplings were carried out to compare 2 L/min, IOM cassettes and 1 or 2 L/min 37-mm closed-face cassettes. The airborne particles were also sampled simultaneously for granulometric analysis. Gravimetric determinations of sampled aerosol were obtained by weighing 37-mm filters and IOM cassettes, and the aerosol collected on the filters and the particles deposited on the walls were analyzed separately for both types of samplers by atomic spectrometry for metals content. The intra-sampler variability and inter-sampler ratios were then determined. Although results obtained for gravimetric analysis are comparable to those published elsewhere (ratio IOM/37-mm much higher than 1), the metal analysis revealed a close agreement between the results obtained with the three sampling methods tested when the wall deposits were taken into account. As published previously, the ratio of wall deposits to filter collected aerosol for 37-mm cassettes is variable, and it would appear to be very difficult to find an appropriate correction factor applicable when only the filter is analyzed. Were these results to be confirmed by further experiments, sampling with 37-mm closed-faced at 1 or 2 L/min or with an IOM sampler would be equivalent for all pollutants for which the analytical method allows the recovery of walls deposit. PMID- 11871757 TI - Respirable concrete dust--silicosis hazard in the construction industry. AB - Concrete is an extremely important part of the infrastructure of modern life and must be replaced as it ages. Many of the methods of removing, repairing, or altering existing concrete structures have the potential for producing vast quantities of respirable dust. Since crystalline silica in the form of quartz is a major component of concrete, airborne respirable quartz dust may be produced during construction work involving the disturbance of concrete, thereby producing a silicosis hazard for exposed workers. Silicosis is a debilitating and sometimes fatal lung disease resulting from breathing microscopic particles of crystalline silica. Between 1992 and 1998, the National Institute for Occupational Safety and Health (NIOSH) made visits to construction projects where concrete was being mechanically disturbed in order to obtain data concerning respirable crystalline silica dust exposures. The construction activities studied included: abrasive blasting, concrete pavement sawing and drilling, and asphalt/concrete milling. Air samples of respirable dust were obtained using 10-mm nylon cyclone pre separators, 37-mm polyvinyl chloride (PVC) filters, and constant-flow pumps calibrated at 1.7 L/min. In addition, high-volume respirable dust samples were obtained on 37-mm PVC filters using 1/2" metal cyclones (Sensidyne model 18) and constant-flow pumps calibrated at 9.0 L/min. Air sample analysis included total weight gain by gravimetric analysis according to NIOSH Analytical Method 600 and respirable crystalline silica (quartz and cristobalite) using x-ray diffraction, as per NIOSH Analytical Method 7500. For abrasive blasting of concrete structures, the respirable crystalline silica (quartz) concentration ranged up to 14.0 mg/m3 for a 96-minute sample resulting in an eight-hour time-weighted average (TWA) of 2.8 mg/m3. For drilling concrete highway pavement the respirable quartz concentrations ranged up to 4.4 mg/m3 for a 358-minute sample, resulting in an eight-hour TWA of 3.3 mg/m3. For concrete wall grinding during new building construction the respirable quartz measurements ranged up to 0.66 mg/m3 for a 191 minute sample, resulting in an eight-hour TWA of 0.26 mg/m3. The air sampling results for concrete sawing ranged up to 14.0 mg/m3 for a 350-minute sample resulting in an eight-hour TWA of 10.0 mg/m3. During the milling of asphalt from concrete highway pavement, the sampling indicated a respirable quartz concentration ranging up to 0.34 mg/m3 for a 504-minute sample, resulting in an eight-hour TWA of 0.36 mg/m3. The results of this work indicate the potential for respirable quartz concentrations involving disturbance of concrete to range up to 280 times the NIOSH Recommended Exposure Limit (REL) of 0.05 mg/m3 assuming exposure for an eight- to ten-hour workday. Considering the aging of the concrete infrastructure in the United States, these results pose a challenge to all who have an interest in preventing silica exposures and the associated disease silicosis. PMID- 11871758 TI - Efficiency of final cleaning for lead-based paint abatement in indoor environments. AB - The effectiveness of procedures used for the final indoor cleaning after active lead-based paint abatement were evaluated in a 830 ft3 test chamber. Dry and wet scraping and dry machine sanding were applied to wooden doors obtained from lead hazard control sites. The airborne particle concentration and size distribution were monitored using a real-time particle size spectrometer. Particulates were also collected on filters and analyzed for total dust and lead. The resulting airborne lead mass was determined for each cleaning procedure, and the potential floor lead loading resulting from the dust settling was calculated. Wipe samples were collected to measure the actual floor lead loading. The effectiveness of final cleaning was evaluated first for dry abatement methods. Various cleaning work practices were tested by applying wet and dry debris sweeping as well as no sweeping in combinations with wet and dry removal of plastic sheeting. Considerable resuspension of leaded particles was detected during dry sweeping: the airborne lead mass increase ranged between 65 and 220 percent. However, this increase did not exceed 22 percent when wet sweeping was applied. Minimal or no resuspension was found when the plastic was folded with leaded debris inside (no sweeping was performed prior to the sheeting removal). During folding activity, the "clean" (uncovered) floor surface may be significantly contaminated with leaded dust from workers' shoes and cleaning tools. The first HEPA vacuuming resulted in a 15- to 20-fold decrease of the airborne lead mass; however, it was not sufficient to reduce the floor lead loading to the U.S. Department of Housing and Urban Development (HUD) clearance level of 40 microg/ft2, as determined by wipe sampling. Wet mopping following the first HEPA vacuuming was proven to be effective to reduce the lead loading significantly below 40 microg/ft2. The second HEPA vacuuming resulted in further reduction of the airborne lead mass concentration. The floor lead loading remained much lower than 40 microg/ft2. These results were confirmed in the tests when using wet scraping. Overall, the HUD-recommended cleaning protocol was found to be sufficient in reducing the floor lead loading below 40 microg/ft2. At the same time, several modifications are proposed in this study to further improve the cleaning effectiveness. PMID- 11871759 TI - Detection of type B influenza virus genes from biopsied gastric mucosa. AB - BACKGROUND: We previously reported the existence of type B influenza genes in the gastric mucosa of three patients. The purpose of the present study was to further characterize gastric mucosal infection with type B influenza virus, in which the natural host is limited to humans. METHODS: Reverse transcription polymerase chain reaction (PCR) and nucleotide sequential analyses were performed to detect type B influenza virus genes in gastric mucosa that was biopsied in the summer, when influenza was not epidemic. Statistical analyses of the clinical backgrounds of the patients were also performed. RESULTS: The nucleoprotein genes of type B influenza virus were detected and confirmed in 37 of 80 patients by sequence analyses of the second PCR products, although they were not detected in the first PCR products. The three patients who were receiving acid suppression therapy who were positive for the influenza virus nucleoprotein gene 37 months previously were found to be positive again. Nucleotide sequential analyses of the nucleoprotein gene revealed two missense mutations from the reference strain, suggesting the characteristics of persistent influenza virus infection. Statistical analyses of the patients' clinical backgrounds showed a correlation between influenza RNA positivity in gastric mucosa and acid inhibition therapy. CONCLUSIONS: These results suggest that influenza viruses infect and probably persist in the gastric mucosa under conditions of acid suppression. PMID- 11871760 TI - The protective effect of catechin on gastric mucosal lesions in rats, and its hormonal mechanisms. AB - BACKGROUND: Catechin, a polyphenol contained in tea (a cup of tea contains approximately 0.1% [w/v] catechin), has various physiological effects. The aim of this study was to investigate the mechanism of the inhibitory effect of catechin on gastric mucosal lesions. METHODS: We studied the effect of catechin on gastric mucosal lesions in rats, using a water immersion restraint (WIR) stress-induced gastric mucosal lesion model. We used crude catechin that contained 52.6% (w/w) ( )-epigallocatechin gallate and 16.7% (w/w) (-)-epicatechin gallate. The rats were randomly divided into three groups; control rats freely drank distilled water, and the remaining rats drank 0.1% (w/v) or 1% (w/v) crude catechin-containing water for 2 weeks. We measured fractional areas of hemorrhagic erosion in the gastric mucosa induced by WIR stress for 4h, compared with findings in the controls. We also employed an isolated rat stomach infusion model and measured gastrin, somatostatin, and histamine in the perfusate to endocrinologically investigate the mechanism underlying the putative protective effect of catechin. RESULTS: Catechin had a significant protective effect against the gastric mucosal lesions induced by WIR stress. Catechin also significantly inhibited the release of gastrin, somatostatin, and histamine. CONCLUSIONS: Catechin confers a protective effect against gastric mucosal lesions, and anti-gastric and anti histaminergic effects may be involved in the mechanism of this effect. PMID- 11871761 TI - Propranolol ameliorates thrombocytopenia in patients with cirrhosis. AB - BACKGROUND: Propranolol causes splanchnic arterial vasoconstriction owing to the unopposed alpha vasoconstriction resulting from the blockade of beta-2 adrenoceptors. It is therefore hypothesized that this drug may cause vasoconstriction in the splenic arterial circulation and, thus, modify the manifestations of hypersplenism, such as thrombocytopenia. The aim of the present study was to test this hypothesis. METHODS: Nineteen patients with cirrhosis and thrombocytopenia (fewer than thrombocytes 7 x 10(4)/mm3) were include. The subjects of the study. All of them were studied in the morning after an overnight fast. To evaluate splenic arterial hemodynamics, the pulsatility index was measured by Doppler ultrasonography. Platelet counts and platelet-associated immunoglobulin G levels were also recorded. The subjects were then randomized to receive propranolol (n = 10) or placebo (n = 9). The measurements were repeated after 1 week of propranolol or placebo administration. The dose of propranolol was determined so that a 20% to 25% reduction in heart rate was achieved. RESULTS: Placebo administration caused no significant changes in splenic artery hemodynamics. In contrast, propranolol administration significantly increased the intra splenic artery pulsatility index (from 1.10+/-0.06 to 1.24+/-0.08; P < 0.01). Placebo administration caused no significant changes in the platelet count. In contrast, propranolol administration significantly increased the platelet count (from 4.5+/-0.3 to 6.1+/-0.73 x 10(4)/mm3; P < 0.05). Furthermore, the change in platelet count was significantly correlated with either the change in extrasplenic artery pulsatility index (r = 0.78, P < 0.05) or the change in intrasplenic artery pulsatility index (r = 0.78, P < 0.01). Platelet-associated immunoglobulin G levels were not modified in either of the two groups. CONCLUSIONS: Propranolol ameliorates thrombocytopenia in patients with cirrhosis. This effect may be caused mainly by hemodynamic changes in the spleen, rather than being caused by immunological mechanisms. PMID- 11871762 TI - Five days of ceftriaxone to treat spontaneous bacterial peritonitis in cirrhotic patients. AB - BACKGROUND: The aim of this study was to determine whether a short course of ceftriaxone was sufficient to cure spontaneous bacterial peritonitis (SBP) in cirrhotic patients. METHODS: We studied 33 cirrhotic patients with SBP. All of them were treated with ceftriaxone, 1.0 g IV, every 12 h for 5 days. Twenty-one variables were recorded to evaluate their relationship to the resolution of SBP. RESULTS: The mean age of the patients was 45 years. Twenty-three were males and 10 females. The etiology of cirrhosis was alcoholic in 42% of the patients, and 82% of the patients belonged to Child-Pugh Class C. Hepatic encephalopathy was present in 39% of the patients. The most frequent organism causing SBP was Escherichia coli (60%). Resolution of SBP on day 5 of treatment was achieved in 73% of the patients. Total resolution of SBP after prolonged therapy with ceftriaxone or another agent. selected according to antibiotic susceptibility, was achieved in 94% of the patients. Hospital mortality was 12%. Multivariate analysis showed no factor that was significantly related to the resolution of SBP, but univariate analysis showed that renal impairment and positive culture tended to be related. CONCLUSIONS: A short course (5 days) of ceftriaxone is useful therapy for SBP. If the polymorphonuclear differential count in ascitic fluid is less than 250 cells/mm3 on day 5 of treatment, the antibiotic can be discontinued. PMID- 11871763 TI - Recurrent nonocclusive mesenteric ischemia after resection of iliac artery aneurysm. AB - A case of recurrent nonocclusive mesenteric ischemia in a patient with isolated internal iliac artery aneurysm penetrating the sigmoid colon is described. On the day after the aneurysm and the sigmoid colon had been resected, the patient developed necrosis of the left hemicolon. Fourteen and nineteen days after left hemicolectomy, massive intestinal bleedings occurred, requiring ileectomy. On the basis of operative findings of good pulsation of visceral arterial branches; angiography showing patent mesenteric vessels with some spasms; and pathological findings suggesting mesenteric ischemia, these ischemic events were diagnosed as nonocclusive mesenteric ischemia. Low-output syndrome induced by massive intestinal bleeding and atrial fibrillation and sepsis were responsible for the establishment of the nonocclusive mesenteric ischemia. Development of disseminated intravascular coagulation and continuous administration of diuretics for acute renal failure seemed to have further perturbed the mesenteric circulation. The patient died of subsequent multiple organ failure 4 months after the first operation. We should pay more attention to nonocclusive mesenteric ischemia in patients with mesenteric ischemia, and strict circulatory management during the perioperative period is essential in these patients. PMID- 11871764 TI - Primary hepatic lymphoma in a patient with Sjogren's syndrome. AB - We report a case of primary hepatic lymphoma (PHL) that developed in a patient with well controlled Sjogren's syndrome (SS) who had been receiving long-term prednisolone treatment. The tumor, found incidentally by ultrasonography, was shown as an extremely large, solitary liver mass that exhibited the immunohistological features of a B-cell lymphoma. An intensive regimen of chemotherapy induced complete regression of this mass. SS occasionally gives rise to lymphoma in salivary glands, however, there have been no previous reports of PHL in a patient with SS. This case study raises the possibility that SS itself, a form of autoimmune disease, and/or prednisolone-based immunosuppressive treatment for SS, predisposes to the occurrence of PHL. SS patients should be monitored vigilantly for the occurrence of extranodal lymphoma, including PHL, even if the SS is well controlled. PMID- 11871765 TI - Autoimmune pancreatitis starting as a localized form. AB - Autoimmune pancreatitis is a rare type of pancreatitis that is characterized by diffuse swelling of the pancreas and irregular stenosis of the pancreatic ductal system ("sclerosing" pantreatitis), caused by autoimmune processes. Previous reports have shown the complete form of the disease, but very few have presented follow-up imagings from the beginning to the complete form. We, herein, report a case of autoimmune pancreatitis starting as a localized form. A-56-year-old Japanese man developed obstructive jaundice. Ultrasonography showed a hypoechoic mass in the head of the pancreas, and endoscopic retrograde pancreatography (ERP) showed localized stenosis of the pancreatic duct in the head of the pancreas. Computed tomography (CT) showed enlargement, with a capsule-like rim, in the head of the pancreas. Internal biliary tube drainage was performed to relieve the obstructive jaundice. The patient was followed-up under the tentative diagnosis of localized "mass-forming" pancreatitis. Four months after the drainage, CT showed diffuse swelling of the pancreas, with a capsule-like rim, and ERP demonstrated diffuse irregular narrowing of the pancreatic duct. Glucose intolerance was noted for the first time. Steroid was given as a diagnostic treatment for autoimmune pancreatitis. Two months after initiation of the steroid treatment, the ERP findings were normal, and CT showed a normal pancreas. The biliary tube was removed, and the glucose intolerance was subsequently alleviated. To summarize, we report a case of autoimmune pancreatitis starting as localized "mass-forming" pancreatitis with a peripheral rim on imagings. It is very important to be well aware of the presence of the localized form of autoimmune pancreatitis. PMID- 11871766 TI - Primary squamous cell carcinoma of the liver producing parathyroid hormone related protein. AB - We report here an autopsy case of primary squamous cell carcinoma of the liver in a 63-year-old man who had hypercalcemia and an elevated serum level of parathyroid hormone-related protein. At autopsy, primary squamous cell carcinoma of the liver was found, without distinct preceding or associated hepatic or biliary diseases; no extrahepatic primary focus of squamous cell carcinoma was found. Bone involvement was not demonstrated, either radiologically or pathologically. Immunohistochemically, parathyroid hormone-related protein was detectable in the squamous cell carcinoma cells and it may have been responsible for the hypercalcemia. Such a case has not been reported so far in the English language or the Japanese literature. PMID- 11871767 TI - Interleukin-2-induced reversible hyperbilirubinemia and cholestasis in a patient with Gilbert's syndrome. PMID- 11871768 TI - Key molecules in metaplastic gastritis: sequential analysis of CDX1/2 homeobox gene expression. PMID- 11871769 TI - Marked improvement in the resolution of, and survival rates in, spontaneous bacterial peritonitis. PMID- 11871770 TI - Helicobacter pylori and gut hormones. AB - Helicobacter pylori infection has been found to decrease the expression of antral somatostatin and to increase the release of the acid-stimulating hormone gastrin. The reversal of these changes in gut hormones by the eradication of H. pylori, and in-vivo and in-vitro studies in animals either infected with H. pylori or exposed to H. pylori-related materials may support the somatostatin-gastrin link theory in the pathophysiology of H. pylori infection. The following mechanisms have been proposed to explain the H. pylori infection-associated changes in gut hormones; (1) ammonia produced by H. pylori and monochloramine, (2) effect on somatostatin receptor subtype-2, (3) action of lipopolysaccharide from H. pylori on somatostatin receptor, (4) inflammatory cells and mediators, and (5) bacterial strain diversity. H. pylori infection can alter gastric acid secretion in both directions. The elevated acid secretion in patients with duodenal ulcer is decreased by H. pylori eradication, and is accompanied by the normalization of gut hormones in patients whose H. pylori-induced gastritis is limited to the antrum with hyperacidity. Corpus gastritis and the subsequent development of mucosal atrophy induced by H. pylori result in decreased acid secretion, although the mechanism underlying H. pylori-induced atrophy in some subjects remains unclear. Hypoacidity enhances corpus atrophy and increases gastrin secretion, mediated via a physiological suppression of somatostatin release, features that are also observed in H. pylori infection. Therefore, the capacity of acid secretion and distribution of gastritis or atrophy should be taken into consideration when we discuss the affect of H. pylori on gut hormones. PMID- 11871771 TI - Postprandial hypotension in elderly subjects: spectral analysis of heart rate variability and electrogastrograms. AB - BACKGROUND: In order to clarify the mechanism of postprandial hypotension in the elderly, the influence of gastric motility and autonomic nervous activity on hypotensive reactions after meals was investigated, using electrogastrograms (EGGs) and spectral analysis of heart rate variability. METHODS: EGGs, heart rate variability, blood pressure, and blood catecholamine levels before and after a meal were measured in 20 healthy young subjects (mean age, 25.6+/-5.6 years; young group) and in 20 healthy elderly subjects (mean age, 78.3+/-5.6 years; elderly group). RESULTS: In the analysis of heart rate variability, no significant changes were observed in the low-frequency component (LF power), high frequency component (HF power), or LF/HF ratio after the meal in the young group. In the other hand, the LF/HF ratio was significantly increased after the meal in the elderly group. In the EGG analysis, the peak power amplitudes after the meal were significantly increased compared with those before the meal in both groups. After the meal, the peak power amplitudes in the young group were significantly greater than those in the elderly group. The baseline blood noradrenaline level (before the meal) was higher in the elderly group than in the younger group, but the level of this catecholamine in the elderly group did not increase significantly after the meal. CONCLUSIONS: It is suggested that the down regulation of catecholamine may be one of the causes of postprandial hypotension in the elderly. The response to secreted catecholamine and the compensatory response to decreased blood flow in the systemic circulation were impaired in the elderly group, which finding may explain the high incidence of postprandial hypotension in the elderly subjects. PMID- 11871772 TI - Expression of homeobox gene CDX2 precedes that of CDX1 during the progression of intestinal metaplasia. AB - BACKGROUND: The CDX1 and CDX2 genes are intestinal transcription factors that may be involved in the regulation of proliferation and differentiation of intestinal epithelial cells. There have been no detailed reports directly comparing the expression of CDX1 with that of CDX2 in chronic gastritis and intestinal metaplasia. Accordingly, we examined the expression of CDX1/2 and its association with the expression of other intestinal metaplasia-associated genes during the development of intestinal metaplasia. METHODS: The expression of CDX1/2 genes was analyzed, using the reverse transcriptase-polymerase chain reaction, in 44 human gastric tissue samples obtained endoscopically. The expressions of mucin markers (MUC2, MUC5AC), intestine-specific genes (sucrase-isomaltase, human defensin-5, alkaline phosphatase), a gene marker for fundic gland area (H+/K+ATPase beta subunit), and a gene for entire gastric glands (pepsinogen C) were also comparatively analyzed. RESULTS: There was no expression of CDX1/2 in gastric mucosa not infected by Helicobacter pylori. The prevalence of CDX1 mRNA expression was significantly higher in mucosa with intestinal metaplasia than in mucosa without intestinal metaplasia. It is noteworthy that CDX2 was expressed in the antral and fundic mucosa in the absence of the expression of CDX1 and gene markers for intestinal metaplasia. CONCLUSIONS: The expression of CDX2 precedes those of CDX1, sucrase-isomaltase, other intestine-specific genes (human defensin 5, alkaline phosphatase), and MUC2 during the progression of intestinal metaplasia. These findings imply that the expression of CDX2 may trigger the initiation and development of intestinal metaplasia. PMID- 11871773 TI - Neuronal cell death, nerve growth factor and neurotrophic models: 50 years on. AB - Viktor Hamburger has just died at the age of 100. It is 50 years since he and Rita Levi-Montalcini laid the foundations for the study of naturally occurring cell death and of neurotrophic factors in the nervous system. In a period of less than 10 years, from 1949 to 1958, Hamburger and Levi-Montalcini made the following seminal discoveries: that neuron cell death occurs in dorsal root ganglia, sympathetic ganglia and the cervical column of motoneurons; that the predictions arising from this observation, namely that survival is dependent on the supply of a trophic factor, could be substantiated by studying the effects of a sarcoma on the proliferation of ganglionic processes both in vivo and in vitro; and that the proliferation of these processes could be used as an assay system to isolate the factor. This work provides a short review mostly of the early history of this subject in the context of the Hamburger/Levi-Montalcini paradigm. This acts as an introduction to a consideration of models that have been proposed to account for how the different sources of growth factors provide for the survival of neurons during development. It is suggested that what has been called the 'social-control' model provides the most parsimonious quantitative description of the contribution of trophic factors to neuronal survival, a concept for which we are in debt to Viktor Hamburger and Rita Levi-Montalcini. PMID- 11871774 TI - Neurokinin-1 receptors in the cerebrovascular vasoactive intestinal polypeptide containing nerves in the rat. AB - Recently, the functions of several putative neurotransmitters such as catecholmines, acetylcholine (ACh) and neuropeptides have been elucidated in the cerebrovasculature. The interaction of such neurotransmitters and their receptors, however, has not been sufficiently clarified. The purpose of this study is to explore the relation of recently demonstrated neurokinin-1 (NK-1) receptor-containing cerebrovascular nerve fibers with the cerebrovascular vasodilatory nerves by means of the sequential-staining immunohistochemical method. Numerous sites of NK-1 receptor immunoreactivities were noted along the nerve fibers with vasoactive intestinal polypeptide (VIP) immunoreactivity in the pial arteries in all regions of the brain. They consisted of fine, delicate varicose fibers and thick bundles without varicosities. After sequential staining with VIP, NK-1 receptor immunoreactive material was demonstrated in the VIP positive nerve fibers. The majority of fibers were positive for VIP alone (75%). The number of fibers positive for both NK-1 receptor and VIP was about one-third the number of fibers for VIP alone (22%). Fibers positive for NK-1 receptor alone comprised a small population (3%). This study demonstrated that NK-1 receptors are localized in axonal membrane of VIP-containing parasympathetic nerves. This suggests that the sensory nerves modulate the functions of parasympathetic nerves in peripheral nervous system, such as those on cerebral vessels. PMID- 11871775 TI - Hyperglycemia impairs antro-pyloric coordination and delays gastric emptying in conscious rats. AB - Delayed gastric emptying has been shown in diabetes. Although it has been proposed that hyperglycemia, and not only autonomic neuropathy, contributes to the pathogenesis of delayed gastric emptying, the inhibitory mechanism of hyperglycemia on gastric emptying remains unclear. We studied the effects of hyperglycemia per se on gastric emptying and postprandial gastric motility in conscious rats. Liquid and solid gastric emptying were compared between saline infused rats and D-glucose-infused rats. Two strain gauge transducers were implanted on the antrum and pylorus and the postprandial antro-pyloric coordination was compared between euglycemia and hyperglycemia. D-glucose infusion for 30 min increased blood glucose level from 5.4 +/- 0.5 to 13.0 +/- 1.3 mM and significantly delayed gastric emptying. Forty minutes after the feeding, contractions with low frequency (<3 cycles min(-1)) and high amplitude (>15 g) of the antrum were observed. This period reflects the emptying process of the gastric content and the coordination between the antrum and pylorus was frequently observed. D-glucose infusion significantly reduced feeding-induced antral contractions and abolished the number of episodes of antro-pyloric coordination. Sham feeding-induced gastric contractions were also significantly reduced by hyperglycemia. Postprandial antro-pyloric coordination was not observed in vagotomized rats, suggesting a mediation of vagus nerve. It is concluded that hyperglycemia impairs antral contractions and antro-pyloric coordination in rats. The inhibitory effect of hyperglycemia on gastric emptying is mediated, at least in part, via impaired vagal activity. PMID- 11871776 TI - Expression of 5-HT3 receptors in primary sensory neurons of the petrosal ganglion of adult rats. AB - By using a specific antiserum, expression of the 5-HT3 receptor was examined in the petrosal ganglion (PG) of adult male rats. We found that the 5-HT3 receptors are widely distributed in the PG. This finding was confirmed by RT-PCR detection of the 5-HT3 receptor mRNA in the tissue. Unlike the distribution patterns of tyrosine hydroxylase (TH), which occurred in limited regions of PG, the 5-HT3 receptors seemed to distribute throughout the ganglion. As many TH-positive neurons in PG innervate type I cells in the carotid body, the coexistence of 5 HT3 receptor and TH in some neurons suggests that this receptor may play a role in carotid body chemoreception. PMID- 11871777 TI - The hepatic vagus nerve in the control of insulin sensitivity in the rat. AB - The objective was to determine if the cervical vagus or hepatic branch of the vagus nerve is a suitable site to produce functional parasympathetic denervation of the liver in the rat as assessed from the ability to produce insulin resistance. Anterior plexus denervation in both anesthetized rats and cats results in insulin resistance as assessed by the rapid insulin sensitivity test (RIST). This diagnostic test is a modified euglycemic clamp using the amount of glucose required to be infused to maintain euglycemia following a bolus administration of insulin (50 mU kg(-1) over 5 min, 0.1 ml min(-1) infusion) as the index of insulin sensitivity. Blood sampling was achieved through an arteriovenous silicone vascular shunt connecting the left carotid artery and the right jugular vein and allowed the close monitoring of blood glycemia throughout the test (every 2 min). The control RIST index (249.2 +/- 10.2 mg kg(-1)) was significantly decreased (P<0.001) following hepatic vagotomy (134.0 +/- 13.9). The intraportal infusion of 2.5 microg kg(-1) min(-1) of acetylcholine partially reversed (202.1 +/- 12.3) the insulin resistance. Intravenous atropine (1 mg kg( 1)) or hepatic anterior plexus denervation did not produce significant further insulin resistance. A similar degree of insulin resistance was produced by bilateral cervical vagotomy which was also partially reversed by acetylcholine. Complete hepatic parasympathetic denervation was achieved by selective hepatic vagal branch section. The data suggest that all of the parasympathetic nerves that regulate hormonal control of insulin resistance pass through the cervical vagus and the hepatic branch, and finally, through the anterior hepatic plexus along the common hepatic artery and that denervation at any of these sites leads to functional elimination of all hepatic parasympathetic input regulating insulin sensitivity. This approach provides an additional research tool to study the hepatic parasympathetic reflex control of peripheral insulin action. PMID- 11871778 TI - Enhanced response of subfornical organ neurons projecting to the hypothalamic paraventricular nucleus to angiotensin II in spontaneously hypertensive rats. AB - Thirty subfornical organ (SFO) neurons in normotensive Wistar-Kyoto (WKY) rats and 32 SFO neurons in spontaneously hypertensive rats (SHR) were antidromically activated by electrical stimulation of the hypothalamic paraventricular nucleus (PVN) under urethane anesthesia. The spontaneous firing rate was significantly higher in SHR than in WKY rats. No significant differences in the latency, conduction velocity, and threshold of antidromic response were observed between WKY and SHR. All the identified SFO units were tested for a response to intracarotid injection of angiotensin II (ANG II, 20-ng/kg b.w.t.). Injections of ANG II elicited an increase in the activity of 21 units in WKY and 20 units in SHR and a depression in the firing of one unit in WKY rats, but did not affect the remaining units. The magnitude of the excitatory response caused by the ANG II injection was much greater in SHR than in WKY rats. These results show that there are differences between WKY and SHR in the spontaneous discharge rate of SFO neurons projecting to the PVN and in their response to circulating ANG II. PMID- 11871779 TI - Inhibition of vasomotion in hippocampal cerebral arterioles during increases in neuronal activity. AB - The activity of small arterioles, internal diameter 9.9 +/- 0.8 microm (SEM), was investigated in the CA1 region of hippocampal slices maintained in vitro at 34 degrees C. Under resting conditions, the vessels were quiescent. However, in the presence of the thromboxane A2 agonist U46619 (75-100 nM), rhythmic contractile activity (vasomotion, 1.1-9.9 min(-1), mean 4.1 +/- 0.7 min(-1) SEM) developed in the smooth muscle cells of the vessel walls. Electrical stimulation of the Schaffer collateral fibre pathway was used to evoke increases in neuronal activity in CA1 in the vicinity of the vessels under investigation. A 3-min period of electrical stimulation of the Schaffer collateral fibre pathway produced a significant reduction in vasomotion in 8/8 vessels. During stimulation, vasomotion either ceased completely (n = 5) or the frequency decreased from 7.1, 3.3 and 3.2 min(-1) to 1.2, 0.4 and 0.6 min(-1), respectively (n = 3). In addition, the amplitude of the residual contractions was reduced by 66%, 12% and 52%. In the presence of 1 microM tetrodotoxin (TTX) (n = 4) to block the generation of action potentials, vasomotion was still present. However, the inhibition of vasomotion evoked by increased neuronal activity was blocked concomitant with the abolition of the field potentials recorded in CA1 in response to the stimulation of the Schaffer collaterals. These findings suggest that a reduction in vasomotion may contribute to the local hyperaemia, which accompanies increases in synaptic activity in the brain. PMID- 11871780 TI - Syncopal attack alters the burst properties of muscle sympathetic nerve activity in humans. AB - This study aimed at examining whether the properties of microneurographically recorded muscle sympathetic nerve activity (MSNA) were altered during hypotensive attacks. A retrospective study was performed on 74 subjects who participated in tilt studies when vasodepressive syncope was induced incidentally in six subjects. The specific features of MSNA that distinguish this activity from skin sympathetic nerve activity are (1) rhythmic pulse synchronous burst discharge, (2) a duration of approximately 150-300 ms, and (3) no response to arousal stimuli were abolished during the syncopal attack. The altered features observed during the syncopal attack in these six subjects were (1) scattered reflex latencies of MSNA peak from the ECG R-wave, (2) elongated burst duration twice to five times as long as that in conscious state, and (3) response to arousal stimuli. The reduced input from the baroreceptors due to suppression on the central sympathetic volley proximal to the nucleus tractus solitarius might be attributed to the lost features characteristic of MSNA. PMID- 11871781 TI - Firing properties of single postganglionic sympathetic neurones recorded in awake human subjects. AB - For over three decades, the technique of microneurography has allowed us to record sympathetic neural outflow directly from postganglionic axons in awake human subjects. But because sympathetic axons are clustered within a nerve fascicle, such recordings have been limited to the analysis of multi-unit neural activity. To improve the information content of intraneural recordings, we developed the single-unit approach, in which focal recordings can be made from a single C-fibre via a high-impedance tungsten microelectrode. In this review, we describe our methodology for analyzing unitary sympathetic activity and discuss the similarities in the firing properties of individual muscle vasoconstrictor, cutaneous vasoconstrictor and sudomotor neurones. PMID- 11871782 TI - Mediation by nucleus tractus solitarii glutamatergic neurotransmission of the cardiovascular reflex evoked by distal esophageal distension. AB - Distention of the rat distal esophagus evokes an arterial pressor and a cardioaccelerator response that depends upon activation of a vagal afferent projection to the nucleus tractus solitarii (NTS). The present study aimed to determine in urethane-anesthetized rats if the afferent limb of this reflex (a) relays in the NTS subdivision known ro receive esophageal afferents, and (b) utilizes glutamatergic synapses. To this end, tetrodotoxin or the glutamate antagonists gamma-D-glutamyl-glycine, 6-7-dinitroquinoxaline-2,3-dione (DNQX) and 2-amino-5-phosphonovaleric acid (AP-5) were applied to the NTS extraventricular surface rostral to the obex. All four agents inhibited both components of the reflex. DNQX or AP-5 produced a similar reversible inhibition upon pressure ejection in the vagal esophageal afferent projection area. Application of tetrodotoxin to the dorsomedial medulla oblongata caudal to the area postrema (AP) was ineffective. Basal heart rate (HR) (except in the case of AP-5) and blood pressure increased upon NTS surface application of the antagonists but not after intra-NTS ejection. At corresponding dorsal NTS sites, focal excitation of solitarial neurons with glutamate evoked hypotension and cardiac slowing. At adjacent ventral sites in the NTS subnucleus centralis (NTSc) and/or its immediate vicinity, glutamate elicited an arterial pressor response that coincided with an esophageal contraction in most but not all cases. In conclusion, afferent fibers of the esophago-cardiovascular reflex (ECVR) probably (1) terminate in the vicinity of esophageal premotor neurons comprising the NTSc and (2) activate second-order neurons via glutamate receptors of both the NMDA and non-NMDA subtype. PMID- 11871783 TI - The pressor effect of NO synthase inhibition correlates to pre-existing systolic BP in the rat. AB - A number of studies have found that the vasopressor effect of nitric oxide (NO) synthase inhibition is small following treatment with hypotensive agents but is enhanced after hypertensive agents, and have implicated NO in the mechanism of action of these drugs. We investigated the hypothesis that the rate of vascular NO synthesis is directly related to blood pressure. The vasopressor effect of 10 mg/kg of L-nitro-L-arginine methyl ester (L-NAME) was studied in relation to changes in BP induced by a variety of treatments in both pentobarbital sodium anesthetized and pithed rats. BP reductions were induced by blood withdrawal, surgery and pithing. BP increases were made by injecting 10 and 15 microg/kg boluses of phenylephrine or by injecting 5% albumin solution. Pithing decreased baseline BP and attenuated the vasopressor effect of L-NAME while phenylephrine increased both BP levels and the hypertensive effect of L-NAME. Volume expansion with 5% albumin solution increased both BP and the vasopressor effect of L-NAME. Both surgery (abdominal incision) and withdrawal of 1 ml blood reduced BP and attenuated the pressor effect of L-NAME. When the results of all these studies were combined, systolic BP was found to correlate strongly with the vasopressor effect of L-NAME (R2 = 0.73, P < 0.0001). Diastolic BP correlated less well with L-NAME (R2 = 0.36, P < 0.0003). The results suggest that shear stress generated by blood flow during the systole releases NO, and lowers BP. The pressor effect of NO synthase inhibition is closely related to pre-existing systolic BP. PMID- 11871784 TI - Effects of brief ischaemia on myocardial acetylcholine and noradrenaline levels in anaesthetized cats. AB - Although brief ischaemic events make the myocardium tolerant to subsequent prolonged ischaemia, known as ischaemic preconditioning, whether brief ischaemia also affects neural regulation at the in vivo heart remains unknown. We examined the effects of brief ischaemia on myocardial interstitial acetylcholine (ACh) and noradrenaline (NA) levels in anaesthetized cats (n = 6). Baseline ACh and NA levels were 0.65 +/- 0.13 and 0.66 +/- 0.17 nmol l(-1) (mean +/- SE), respectively. Two sets of 5-min brief occlusion followed by 20-min reperfusion of the left anterior descending coronary artery (LAD) significantly increased the myocardial interstitial ACh level to 4.6 +/- 0.7 nmol l(-1) (P < 0.01), while not affecting the myocardial interstitial NA level. Subsequent 60-min LAD occlusion significantly increased the ACh and NA levels to 34.9 +/- 6.0 and 96.5 +/- 17.0 nmol l(-1) (P < 0.01), respectively. Vagotomy abolished the myocardial interstitial ACh release during brief ischaemia and attenuated the ACh release during subsequent 60-min ischaemia (n = 6). In contrast, vagotomy did not affect the subsequent ischaemia-induced myocardial interstitial NA release. We conclude that the brief ischaemia affects myocardial interstitial ACh release but not NA release in the ischaemic myocardium in vivo. PMID- 11871786 TI - Distribution of sympathetic preganglionic neurons innervating the kidney in the rat: PRV transneuronal tracing and serial reconstruction. AB - The organization of spinal motor circuitry to the kidney is not well characterized and changes in renal innervation have been associated with disease states such as hypertension found in the spontaneously hypertensive rat or renal hypertension. Here, we describe the segmental and intra-segmental organization of the spinal motor circuitry that was resolved after neurotropic viral injection into the kidney and retrograde transneuronal transport to the spinal cord. In the first experiment, the serial reconstruction of infected neurons in the thoracolumbar spinal cord from T8-L1 was performed following injection of pseudorabies virus (PRV, Bartha strain) into either the cranial pole, the caudal pole or both the cranial and caudal poles of the left kidney in male rats. In the second experiment, rats received injections of two different PRV strains that were genetically engineered to express unique reporter molecules; one of the engineered strains was injected into the cranial pole and the other was injected into the caudal pole. Either 3- or 4-day post-infection, the animals were anesthetized and sacrificed by transcardial perfusion. PRV-infected neurons were located by immunocytochemistry against either PRV itself (experiment 1) or the unique marker proteins (experiment 2). After injection of both poles of the kidney, the majority of the infected neurons were found in the ipsilateral intermediolateral cell column (IML) from T10 to T12 with the mode at T11. Infected neurons were found in discrete neuron clusters in the intermediolateral cell column along the longitudinal axis in a repeating pattern of high and low density that has been called "beading". Three observations indicated a topographic distribution of renal sympathetic preganglionic neurons (SPN). First, after injection into either the cranial or caudal poles of the kidney, the mode of infected cells was located in segments T11 and T12, respectively. The one spinal segment shift in the mode suggested a topographic distribution. Second, in spinal segments T8-L1, comparison of the distributions of the neurons innervating each pole of the left kidney revealed an overlap in the distribution, except in the T11 segment. In the T11 segment, the neurons projecting to each pole tended to segregate into separate populations. Third, in rats that received injections of two PRV strains that were genetically engineered to express unique markers into opposite poles of the kidney, a segregation of neurons projecting to the cranial and caudal poles of the kidney was noted again in the T11 spinal segment and the segregation at adjacent spinal levels was obvious. The analysis of the distribution of infected neurons within each spinal cord segment (intra-segmental distribution) revealed three different patterns along the cranial-caudal dimension. In segments T8-T10, >60% of the infected neurons were located in the caudal half of the spinal segment. In segments T12-L1, >60% of the infected neurons were located in the cranial half of the spinal segment. In segment T11, the neurons were more evenly distributed throughout the segment. These intra segmental distribution patterns were found after both 3- or 4-day survival periods post-infection and were found in most animals. The distribution of clusters of neurons revealed a similar intra-segmental pattern. Thus, as was described previously for the sympathetic postganglionic neurons that innervate the kidney, the present work indicates a topographic organization in the second order neurons in the renal sympathetic efferent pathway. The physiological significance of this anatomical organization remains to be determined. PMID- 11871785 TI - Neuronal organization and cell interactions of the cat stellate ganglion. AB - The functional structure of the cat stellate ganglion (SG) and, in particular, its extra- and intraganglionic connections and neuronal organization, were investigated using histochemical, immunohistochemical, morphological and histological methods. Retrograde axonal transport of horseradish peroxidase was used to determine most of the extraganglionic interactions. Of the targets tested, the most extensive efferent connections of the SG were with the stemocleidomastoid muscle, trachea, esophagus and heart. Neurons of the SG also send a small number of postganglionic efferents to the thyroid and stomach. Furthermore, ganglion cells send axons to the spinal ganglia. Several afferent connections of the SG were determined. Sympathetic preganglionic neurons of segments C8-T10 of the spinal cord, sensory neurons of C8-T9 spinal ganglia, intramural ganglia of the thoracic viscera and the reticular formation of the medulla oblongata send their axons to the SG. Intraganglionic interactions of intemeurons with principal ganglionic cells were assumed to occur, based on the presence of interneurons immunoreactive to GABA and substance P. GABA- and substance P-immunoreactive fibers located around a small number of postganglionic neurons were also identified. Morphological study revealed asymmetry between the left and right ganglia: the right ganglion is larger than the left and contains more cells. This asymmetry was also reflected in basic structural parameters of neurons, such as average neuronal area and average diameter of cell somata. The present data has been used to develop a scheme for the basic inter- and intraneuronal connections of the cat SG. This ganglion is a true nervous center, with postganglionic neurons, some of which might be performing sensory functions, and interneurons. The ganglion is connected not only with the spinal cord and spinal ganglia, but also with neurons of the intramural ganglia and, by direct links, with efferent neurons of the medulla oblongata. Thus, the SG may play an essential role in viscera-visceral reflexes. PMID- 11871787 TI - Changes in mouse sudomotor function and sweat gland innervation with ageing. AB - Age-related changes in sudomotor neuroeffector function have been evaluated in mice aged 2 (young), 6, 12 (adult) and 18 (old) months. We evaluated sudomotor function by determining the number of sweat glands reactive to pilocarpine and the sweat output per gland on the plantar surface of the hindpaws with the impression mould technique. Protein gene product 9.5 (PGP) and vasoactive intestinal polypeptide (VIP) were immunohistochemically localised in footpads. A marked decrease (44%) in sweat output per gland was observed in old mice as well as a slight (17%), not significant decline in the number of secreting sweat glands. The sudomotor innervation, expressed as the area of sweat gland occupied by VIP and PGP immunoreactive nerve profiles, showed an initial increase from 2 to 6 months and a significant decline (35%) in 18- vs. 6-month-old mice. These results indicate that, in contrast to the number of secreting sweat glands, sweat output per gland does not reach the maximum in adult mouse until 6 months old and that sweating decreases in aged mice mainly due to a decline of sweat output per gland and to a lesser extent to a decrease in number of secreting glands. A reduction of sweat glands size in aged mice was also found, suggesting that the diminished sweat gland responsiveness with ageing may be attributed to sweat gland atrophy as well as to loss of innervation. PMID- 11871788 TI - Intravenous amiodarone modifies autonomic balance and increases baroreflex sensitivity in conscious rats. AB - Amiodarone is an antiarrhythmic agent commonly used to treat cardiac arrhythmias. This study was designed to investigate the effects of intravenous amiodarone on the neural control of heart rate and arterial pressure and spontaneous baroreflex sensitivity (BRS). Experiments were carried out on conscious freely moving normotensive Wistar (WR) and spontaneously hypertensive rats (SHR). Arterial pressure was continuously monitored before and after amiodarone (50 mg/kg i.v.) or vehicle for 30 min. Heart rate (expressed as the pulse interval, PI) and systolic arterial pressure (SAP) variabilities were assessed using autoregressive spectral analysis. BRS was calculated as the alpha-index (the square root of the ratio between the PI and SAP powers). Amiodarone induced bradycardia and hypotension in both strains, with these effects being more intense in SHR. The variability profile of PI was characterized by a significant reduction of normalized low frequency (LF) and LF/HF ratio, while the high frequency (HF) component both in absolute and normalized units (nu) was increased in both WR and SHR strains. A significant decrease in SAP variance and its LF oscillation was observed. In addition, BRS was also increased in both groups, being more intense in SHR. In both WR and SHR, intravenous amiodarone had a considerable effect on heart rate variabilities (HRV), shifting cardiac sympathovagal balance toward a sympathetic inhibition and/or vagal activation, which were associated with an increase in spontaneous BRS. Decreases of SAP variance and LF(SAP) suggest sympatholytic effects on peripheral vessels. Besides the direct ion channel effects, these changes in the autonomic balance could contribute to the antiarrhythmic action of the intravenous amiodarone. PMID- 11871790 TI - Too much learning is a dangerous thing. PMID- 11871789 TI - Volume loading inhibits baroreflex vagal bradycardia in rats. AB - Volume loading tends to increase blood pressure. Resultant arterial baroreflexes, especially the heart rate component, would counteract renal compensation of volume expansion. In this study, we investigated effect of volume loading on baroreflex vagal bradycardia (BVB) induced by electrical stimulation of the aortic depressor nerves in chloralose/urethane-anesthetized, succinylcholine immobilized, artificially ventilated rats. As a result, we confirmed that volume loading inhibits BVB. The inhibition was abolished following spinal cord transection at C2. In contrast, volume loading did not affect the bradycardia induced by electrical stimulation of peripheral cut ends of the cervical vagus. These findings suggest that the inhibition of BVB by volume loading is mediated by afferent input which is conveyed in the sympathetic nerves and ascends in the spinal cord, and produced at a central site. PMID- 11871792 TI - Haemorrhagic viruses as bioweapons. PMID- 11871791 TI - Anthrax attack in the USA. PMID- 11871793 TI - USA to increase smallpox vaccine stockpile. PMID- 11871794 TI - Therapeutic HIV vaccine from Spain. PMID- 11871795 TI - Human trials start for new tuberculosis vaccine. PMID- 11871797 TI - Mexican vaccine against typhoid fever in phase I trial. PMID- 11871796 TI - New initiatives to combat cervical cancer in India. PMID- 11871798 TI - Antibiotic-resistant bacteria in food animals on the rise. PMID- 11871800 TI - Are we ready for the next flu pandemic? PMID- 11871801 TI - Shaping travel health and medicine for the future. PMID- 11871802 TI - The future of polio eradication. AB - As the global polio eradication initiative comes ever closer to its goal of terminating of all wild poliovirus transmission, significant challenges remain. Wild poliovirus transmission must be terminated in countries where low-level transmission persists, in large reservoir countries with high population density, and in conflict countries. Eradication can be achieved in these countries with determined and persistent effort, assuming that sufficient resources are mobilised. High quality surveillance needs to be implemented in the remaining polio endemic countries, especially in Africa. Surveillance is necessary in all countries until eradication is certified. A strategy for stopping immunisation after eradication is yet to be defined. A definition of this strategy would address the emergence of feral polioviruses, chronic vaccine virus infection in immunodeficient persons, and containment of laboratory strains. Until immunisation is stopped, high-population immunity should be maintained through uniformly high immunisation coverage with potent vaccines. PMID- 11871803 TI - Enteroaggregative Escherichia coli. AB - Enteroaggregative Escherichia coli (EAEC) are an increasingly important cause of diarrhoea. E. coli belonging to this category cause watery diarrhoea, which is often persistent and can be inflammatory. EAEC have been implicated in sporadic diarrhoea in children and adults, in both developing and developed countries, and have been identified as the cause of several outbreaks worldwide. EAEC are defined by their ability to adhere to epithelial cells in a characteristic "stacked-brick" pattern but are otherwise highly heterogeneous. Genes that could contribute to the pathogenicity of EAEC encode adhesins, toxins, and other factors, all of which are only partially conserved. Practicable tools are needed to improve diagnosis and identify risk factors. EAEC-infected individuals can be treated with fluoroquinolones but there is a need to examine alternative treatment protocols. PMID- 11871804 TI - Vancomycin-resistant enterococci: why are they here, and where do they come from? AB - Vancomcyin-resistant enterococci (VRE) have emerged as nosocomial pathogens in the past 10 years, causing epidemiological controversy. In the USA, colonisation with VRE is endemic in many hospitals and increasingly causes infection, but colonisation is absent in healthy people. In Europe, outbreaks still happen sporadically, usually with few serious infections, but colonisation seems to be endemic in healthy people and farm animals. Vancomycin use has been much higher in the USA, where emergence of ampicillin-resistant enterococci preceded emergence of VRE, making them very susceptible to the selective effects of antibiotics. In Europe, avoparcin, a vancomycin-like glycopeptide, has been widely used in the agricultural industry, explaining the community reservoir in European animals. Avoparcin has not been used in the USA, which is consistent with the absence of colonisation in healthy people. From the European animal reservoir, VRE and resistance genes have spread to healthy human beings and hospitalised patients. However, certain genogroups of enterococci in both continents seem to be more capable of causing hospital outbreaks, perhaps because of the presence of a specific virulence factor, the variant esp gene. By contrast with the evidence of a direct link between European animal and human reservoirs, the origin of American resistance genes remains to be established. Considering the spread of antibiotic-resistant bacteria and resistance genes, the emergence of VRE has emphasised the non-existence of boundaries between hospitals, between people and animals, between countries, and probably between continents. PMID- 11871805 TI - The role of nuclear medicine in infection and inflammation. AB - Investigators have used various techniques and radionuclides such as 51Cr and 32P diisofluorophosphate to label blood cells and to study cell survival. Early studies also used these radionuclides to label human leucocytes for cell survival by in-vitro counting. But external imaging could not be done with these agents. Starting with the use of the gamma-emitting radionuclide (111)In-oxine for in vitro labelling of phagocytic leucocytes, external imaging became possible. This method was the basis of visualisation of cell distribution within the body. Because an abscess consists primarily of leucocytes, leucocytes labelled with (111)In localise within the abscess and are detectable by imaging. Nowadays other radiopharmaceuticals with other underlying uptake mechanisms are also used to detect inflammatory or infectious foci in patients. Nuclear medicine can be most useful in patients with fever of unknown origin, where a focus has to be defined, or in patients where a lesion is known by clinical symptoms or by a radiological imaging and the differentiation between infection and other pathologies has to be made. PMID- 11871806 TI - Emerging role of Mycoplasma pneumoniae and Chlamydia pneumoniae in paediatric respiratory-tract infections. AB - Increased use of specialised diagnostic techniques over the past 10 years has allowed considerable new information to be obtained concerning Mycoplasma pneumoniae and Chlamydia pneumoniae infections. In children, these pathogens seem to have a more important role in causing respiratory-tract infections than previously thought; they have been associated with wheezing, and they are also frequent in children aged under 5 years. Contrary to original belief, no clinical, laboratory, or radiological findings seem to be unique to M. pneumoniae or C. pneumoniae; furthermore, there is no rapid and cost-effective diagnostic test capable of identifying these pathogens. Appropriate antimicrobial treatment of the infections they cause is needed to reduce the recurrent episodes of wheezing and other respiratory symptoms, to decrease morbidity, and to avoid the spread of the pathogens. However, a number of therapeutic issues remain unsolved. PMID- 11871807 TI - Hot spots in a wired world: WHO surveillance of emerging and re-emerging infectious diseases. AB - The resurgence of the microbial threat, rooted in several recent trends, has increased the vulnerability of all nations to the risk of infectious diseases, whether newly emerging, well-established, or deliberately caused. Infectious disease intelligence, gleaned through sensitive surveillance, is the best defence. The epidemiological and laboratory techniques needed to detect, investigate, and contain a deliberate outbreak are the same as those used for natural outbreaks. In April 2000, WHO formalised an infrastructure (the Global Outbreak Alert and Response Network) for responding to the heightened need for early awareness of outbreaks and preparedness to respond. The Network, which unites 110 existing networks, is supported by several new mechanisms and a computer-driven tool for real time gathering of disease intelligence. The procedure for outbreak alert and response has four phases: systematic detection, outbreak verification, real time alerts, and rapid response. For response, the framework uses different strategies for combating known risks and unexpected events, and for improving both global and national preparedness. New forces at work in an electronically interconnected world are beginning to break down the traditional reluctance of countries to report outbreaks due to fear of the negative impact on trade and tourism. About 65% of the world's first news about infectious disease events now comes from informal sources, including press reports and the internet. PMID- 11871808 TI - Peter George Smith--bridging the gap between epidemiology and research. PMID- 11871809 TI - Natural might not be healthier. PMID- 11871810 TI - Ischemia-reperfusion injury of the liver with special reference to calcium dependent mechanisms. AB - Ischemia-reperfusion (I/R) injury is a complex process involving numerous intracellular signaling pathways, mediators, cells, and pathophysiological disturbances; its prevention during liver surgery is of utmost importance. In this review, we divide hepatic I/R injury into two phases, intracellular and extracellular, for a better understanding of the processes involved. Ca2+ and Ca2+-dependent reactions play an important role as a trigger in the former phase, while the subsequent generation of bioactive substances plays a predominant role in the latter phase. These findings indicate that a combination of different therapeutic approaches against Ca2+-dependent steps may help prevent I/R injury of the liver. PMID- 11871811 TI - Postoperative follow-up of patients with early breast cancer: reappraisal of serum tumor markers. AB - The purpose of this study was to determine the most appropriate tests and procedures to detect disease progression effectively during the postoperative follow-up of patients with early breast cancer. We reevaluated our current surveillance protocol which involves the intensive follow-up of 643 patients with stage I disease. With the exception of one case of bone metastasis, all cases of recurrence (97%) were suspected from abnormal results detected during surveillance involving physical examination, serial determination of tumor markers, and chest roentgenography. Among 15 patients with asymptomatic distant metastasis, disease recurrence was suspected in 12 (80%) because of increased levels of serum tumor markers. No disease recurrence was detected by routine complete blood counts or automated chemistry studies alone. Our experience indicates that an effective follow-up regimen for patients with early breast cancer may include careful history-taking, physical examination, and the determination of serum tumor markers every 3-6 months for the first 3 years, then less frequently thereafter, and chest roentgenography every 6 months for 5 years, in addition to annual mammography. Serial determination of the tumor markers tumor polypeptide antigen, NCC-ST-439, and either carcinoembryonic antigen or carbohydrate antigen 15-3, seems to be of value for the selection of patients who should undergo radiologic exploration. The health benefits and cost-effectiveness of a follow-up focused on the measurement of serum tumor markers need to be evaluated in large prospective randomized trials. PMID- 11871812 TI - Video-assisted thoracoscopic surgery in the treatment of chronic empyema thoracis. AB - We prospectively analyzed the surgical results in chronic organizing empyema thoracis utilizing a video-assisted thoracoscopic technique, particularly in debilitated patients. From January 1999 to September 2000, ten patients with stage III empyema thoracis underwent video-assisted thoracoscopic surgery for decortication (VATD). The mean age of the patients was 53.2 years, and they included one female and nine male patients. Four patients were regarded as not suitable for open thoracotomy. After the procedure, all patients had one infusion tube and two 32-F chest tubes inserted, the former for irrigation with saline solution and the latter for drainage. The mean operation time was 178 min. There was no mortality. The mean time to remove the infusion tube was 3.3 days. The mean time to remove the first chest tube was 7.1 days and the second chest tube 9.7 days. The mean hospitalization time after the operation was 14.9 days. The mean follow-up was 14.9 months. A restoration of lung function was ascertained in nine patients, with a 17.6% mean increase in forced vital capacity. There was no recurrence of empyema during the follow-up. Nevertheless, a reaccumulation of pleural effusion occurred in one patient 1 month after the procedure, which thus necessitated further treatment. VATD is considered to be a feasible surgical modality for the treatment of stage III empyema thoracis in selected patients. PMID- 11871813 TI - Efficacy of a pedicled pericardial fat pad fixed with fibrin glue on postoperative alveolar air leakage. AB - The purpose of this randomized trial was to investigate the effect of using a pedicled pericardial fat pad fixed with fibrin glue on postoperative alveolar air leakage. Thirty consecutive patients with lung cancer, who had moderate alveolar air leaks after pulmonary resection, were randomized into two groups: in group A fibrin glue was applied onto the surface of the leaking raw lung and in group B, after applying fibrin glue in the same manner as in group A, a pedicled pericardial fat pad was immediately fixed to the leaking lung surface with fibrin glue. The duration of the postoperative air leakage and chest tube drainage was recorded. In 6 of 15 patients in group B the air leakage ceased within the first 24h after pulmonary resection, while in group A only 1 of 15 patients showed a cessation of the air leakage, and a significant difference was noticed between the two groups (P = 0.0309). The duration of the postoperative air leakage was 4.8+/-4.6 days in group A and 3.6+/-3.4 days in group B. The pedicled pericardial fat pad fixed onto the surface of the leaking raw lung using fibrin glue was found to reduce alveolar air leakage after pulmonary resection. PMID- 11871814 TI - Role of interleukin-1alpha and type I interleukin-1 receptor in the growth activity and invasion of gastric carcinoma cells. AB - To clarify the role of the interleukin (IL)-1/IL-1 receptor system in the progression of gastric carcinoma cells in patients with advanced gastric cancer, we measured the tissue concentrations of IL-1alpha, the expression of IL-1 receptor type I (IL-1RtI) on tumor cells, and the cell-growth activity through an analysis of DNA content. The concentrations of IL-1alpha were significantly higher in differentiated than in undifferentiated tumors (P = 0.038). The expression of IL-1RtI was upregulated in the tumor cells associated with INFgamma [corrected], scirrhous type tumors, and T3 and T4 tumors. There was a clear linear correlation between the tissue concentrations of IL-1alpha and S-phase fractions in differentiated tumors (r = 0.664, P = 0.003). Tumor cells with high IL-1alpha concentrations and low IL-1RtI expression had significantly greater S phase fractions than those with low IL-1alpha concentrations, independent of IL IRtI expression (P = 0.024 in low IL-1RtI, P = 0.019 in high IL-IRtI). These findings indicate that IL-1alpha stimulates the growth of differentiated gastric carcinoma cells and that IL-IRtI expression is involved in tumor invasive activity. High S-phase levels were not necessarily associated with a high expression of IL-1RtI, which may be due to the downregulatory effects of high IL 1alpha concentrations. PMID- 11871815 TI - Prognostic significance of the cytologic features of free cancer cells in the peritoneal cavity of patients with gastric cancer. AB - The aim of this study was to determine whether the cytologic features of cancer cells in the peritoneal lavage fluid from patients undergoing surgery for gastric cancer affect prognosis. A total of 141 patients with stage III-IV gastric cancer were enrolled in this study. Among 111 patients without peritoneal metastasis (P0), 43 were noted to have positive peritoneal cytology (P0/CY1). These patients were subclassified into two groups according to the cluster formation of cancer cells in the cytology specimen, namely, 17 with cluster formation and 26 without cluster formation. The clinicopathologic parameters and survival rates were compared between these two groups. Among the P0/CY1 patients, no significant difference in clinicopathologic features existed between patients with or without the cluster formation of cancer cells. However, the survival rate of patients without cluster formation was significantly higher than that of those with cluster formation. Furthermore, the survival curves of patients without cluster formation was similar to that of the P0/ CY0 patients with stage IV cancer. Subclassification according to the presence or absence of the cluster formation of cancer cells may be useful for predicting the outcome of patients with positive cytology after surgery for advanced gastric cancer. PMID- 11871817 TI - Control of intraoperative bleeding during liver resection: analysis of a questionnaire sent to 231 Japanese hospitals. AB - To determine the safest and most efficient way of performing hepatectomy, the differences in methods employed by Japanese surgeons were examined. In November 1998, a questionnaire on bleeding control during hepatectomy was sent to 270 hospitals located throughout Japan. The answers from 231 hospitals (85.6%) were analyzed. Surgical apparatus such as an ultrasonic dissector (USD) was used in 203 hospitals. Pringle's maneuver was performed routinely in 25%, for segmentectomy and subsegmentectomy in 25%, for lobectomy in 9%, depending on the situation in 34%, and never in 7%. In 135 hospitals (60%), hemostatic materials such as fibrin glue were always applied to the cut surface after hepatectomy. The USD was chosen and widely accepted by the hospitals studied. As Japanese patients with hepatoma often have liver cirrhosis, intermittent occlusion and the selective clamping of hepatic inflow were considered preferable to persistent inflow occlusion. The gentle exposure of hepatic venous branches, careful hemostasis during hepatectomy, and accurate location of the hepatic vein by intraoperative ultrasonography were all considered to be extremely important. PMID- 11871816 TI - A prospective randomized comparison between an open hemorrhoidectomy and a semi closed (semi-open) hemorrhoidectomy. AB - A semi-closed hemorrhoidectomy is a popular surgical procedure among Japanese coloproctologists because it is thought that the risk of postoperative bleeding is reduced, and postoperative pain is milder after a semi-closed hemorrhoidectomy than after an open hemorrhoidectomy. However, no prospective randomized trial comparing an open and semi-closed hemorrhoidectomy has yet been published. We conducted a prospective randomized trial comparing both clinically and physiologically an open and semi-closed hemorrhoidectomy. Thirty-four consecutive patients undergoing a hemorrhoidectomy for third-degree hemorrhoids were randomized to receive either an open hemorrhoidectomy (n = 17) or a semi-closed hemorrhoidectomy (n = 17). Postoperative pain was evaluated using an analog scale by the patients themselves. An anorectal physiological study was performed before the operation and 2 months after the operation. Pain at 1 week after operation was significantly more severe after a semi-closed hemorrhoidectomy than after an open hemorrhoidectomy. The postoperative physiological parameters including sphincter pressures did not differ between the two forms of hemorrhoidectomy. However, younger patients and patients having higher sphincter pressures preoperatively had more severe pain at 2 weeks after a semi-closed hemorrhoidectomy. Although both forms of hemorrhoidectomy appear to be almost equivalent, the degree of early postoperative pain may be less after an open hemorrhoidectomy in both young patients and in those patients having high preoperative anal sphincter pressures. PMID- 11871818 TI - A novel suicide gene therapy system for p53-mutated cells using a wild-type p53 specific promoter and Cre/loxP switch. AB - Approximately half of all malignant tumors have a mutation or deficiency in the p53 tumor suppressor gene. The present study was designed to develop a p53 mutated cancer cell-specific suicide gene therapy system using p53 as a transcriptional activating factor. We introduced the promoter containing the wild type p53-specific binding sequence (p53SP) and the Cre/loxP exchange system to induce specific expression of the herpes simplex viral thymidine kinase (HSV-tk) gene in p53-mutated cells. The transfection of an enhanced green fluorescent protein (EGFP) reporter gene expression vector containing p53SP resulted in selective EGFP expression in wild-type p53-producing cells, but not in p53 mutated cells. To express HSV-tk alone in the p53-mutated cells, two plasmid vectors were prepared: one regulator plasmid vector to express Cre under the control of the p53SP promoter (p53Cre), and another tk expression vector driven by the CAG promoter containing loxP sites at both ends of the HSV-tk gene (pCALtkL). The cotransfection of p53Cre with pCALtkL preferentially reduced the expression of HSV-tk in the wild-type p53-producing cells, but not in the p53 mutated cells. This cotransfection led to selective growth inhibition in the cotransfected p53-mutated cells mediated by ganciclovir, whereas a single transfection of pCALtkL caused nonselective growth inhibition in both cell lines following ganciclovir treatment. Thus, the HSV-tk expression system containing the wild-type p53-specific promoter and the Cre/loxP switch endabled the selective growth inhibition of p53-mutated cancer cells. PMID- 11871819 TI - Brachiocephalic vein thrombus of papillary thyroid cancer: report of a case. AB - Differentiated thyroid cancer sometimes shows microscopic vascular invasion but rarely causes a tumor thrombus in a great vein. A preoperative diagnosis of the latter condition is difficult if a patient does not have any overt symptoms, but it is important for operative planning. We encountered a 26-year-old woman with papillary thyroid cancer and a tumor thrombus extending from the brachiocephalic vein to the superior vena cava. The tumor was successfully treated by surgical resection. To our knowledge only 19 such cases have so far been reported. We herein report our case, review the literature, and discuss the associated diagnostic problems and treatments including such areas as age, gender, symptoms, pathology, extension, and outcome. PMID- 11871820 TI - Pregnancy after undergoing the Fontan procedure for a double outlet right ventricle: report of a case. AB - We describe a 25-year-old woman who had a successful pregnancy after the repair of a double outlet right ventricle using the Fontan procedure. Although the number of reported patients is small, women who have good hemodynamic results after the operation can apparently tolerate both pregnancy and delivery. PMID- 11871821 TI - Congenital sternal cleft with patent ductus arteriosus: report of a case. AB - We report a rare case of a congenital sternal cleft. The patient was a full-term baby girl with a superior incomplete sternal cleft with patent ductus arteriosus (PDA). A primary repair of the sternum and ligation of the PDA were performed during the neonatal period without cardiac compression. Primary repair during the neonatal period is the optimal procedure for cases of congenital sternal cleft. PMID- 11871822 TI - Small intestinal bleeding, secondary to metastatic malignant fibrous histiocytoma of the rib: report of a case. AB - Malignant fibrous histiocytoma (MFH) is an aggressive bone tumor with a high propensity for metastasis. We describe herein the case of a 73-year-old man found to have MFH of the rib with a metastasis in the small intestine. The patient complained of gastrointestinal bleeding as the first symptom. Computed tomography imaging demonstrated a tumor in the left chest wall, and abdominal magnetic resonance imaging revealed thickening of the wall in the internal cavity of the small intestine. Histopathological and immunohistochemical examination confirmed that the tumor in the small intestine was an MFH that had metastasized from the rib. PMID- 11871823 TI - Long-term survival in advanced small cell carcinoma of the colorectum: report of a case. AB - A 47-year-old man was referred to our institution with bloody diarrhea. An endoscopic examination showed a 6-cm tumor with central ulceration in the upper rectum. A high anterior resection of the rectum with lymphadenectomy was performed with the diagnosis of colorectal cancer. An intraoperative cytological examination found many free cancer cells in the peritoneal lavage fluid. Histologically, the tumor had invaded deeply and exposed the serosa, and was diagnosed as a small cell carcinoma of the colorectum with marginal lymph node metastasis. The patient is alive without any evidence of recurrence approximately 37 months after surgery despite the aggressive clinical behavior and a high mortality rate associated with this tumor. This case of an advanced small cell carcinoma of the colorectum showed a good outcome even though the cytology of the peritoneal lavage was positive. PMID- 11871824 TI - Middle hepatic vein reconstruction using a peritoneal patch: report of a case. AB - A 67-year-old male complaining of constipation with a change in stool caliber for several months visited our hospital in June 1999. A positive test for occult blood in the feces led to the disclosure of a type II carcinoma of the sigmoid colon with multiple liver metastases. A lymph node dissection with a sigmoidectomy disclosed no metastases histologically, so a left hepatectomy and enucleations of the metastases were performed. In addition, the invaded middle hepatic vein (MHV) was resected and repaired using a peritoneal patch. The patient's postoperative course was uneventful until July 2000, when computed tomography of the liver showed a single nodule measuring 3 cm in diameter in segment 6. The metastasis was excised in August. Since then, the patient has shown normal tumor marker values. The MHV has remained patent for 24 months after its reconstruction. A resection of the liver metastases including venous reconstruction is beneficial for patients since it results in a longer survival and allows for venous drainage of the residual liver. The peritoneum is also accessible, enabling the fitting of a patch graft for hepatic vein repair. PMID- 11871825 TI - Inflammatory malignant fibrous histiocytoma of the gallbladder: report of a case. AB - We describe herein a case of inflammatory malignant fibrous histiocytoma (IMFH) of the gallbladder that subsequently metastasized to the ascending colon and later to the stomach. A 70-year-old Japanese man with a palpable mass in the right upper quadrant of the abdomen was referred to our hospital for investigation and treatment. Laboratory data showed severe leukocytosis and elevated serum granulocyte colony-stimulating factor (G-CSF) concentrations. A laparotomy was performed, and the tumor was excised en bloc with the gallbladder and part of the liver bed. Histopathologically, the tumor was composed of ordinary malignant fibrous histiocytoma (MFH) components characterized by pleomorphic tumor cells, bizarre giant cells, and conventional spindle cells in a storiform growth pattern, as well as a xanthogranulomatous component, including inflammatory cells, foamy histiocytes, and plasma cells. Immunohistochemical study revealed that the pleomorphic tumor cells and bizarre giant cells were positive for antibodies against alpha1-antitrypsin and alpha1-antichymotrypsin. The final pathologic diagnosis was IMFH. The tumor cells were diffusely positive for anti-G-CSF monoclonal antibody, and the inflammatory reaction subsided immediately after tumor resection, strongly suggesting that the primary tumor cells produced G-CSF. This patient is still alive with no signs of recurrence more than 3 years after his primary operation, which to our knowledge is the longest survival period ever reported. Therefore, visceral IMFH is manageable in some cases by resecting the primary and isolated metastatic lesions. PMID- 11871826 TI - Ascariasis-induced empyema of gallbladder: report of a case. AB - A rare case of a 52-year-old woman with empyema of the gallbladder due to ascariasis causing an obstruction in the cystic duct is presented. She was admitted on September 20, 2000, and on September 23 an emergency cholecystectomy was performed. Ultrasonography is a highly sensitive and specific method for diagnosing gallbladder ascariasis, and a cholecystectomy is considered mandatory for the treatment of empyema of the gallbladder. PMID- 11871827 TI - Gallbladder metastasis of renal cell carcinoma: report of two cases. AB - We report two extremely rare cases of metastasis to the gallbladder from renal cell carcinoma. In both men, aged 63 and 80 years, a pedunculated polypoid gallbladder tumor was incidentally found 27 and 8 years after surgery for renal cell carcinoma, respectively. The tumors showed hypervascularity on diagnostic imaging. A histopathological examination showed no tumor cells in the gallbladder mucosa. but clear cell carcinoma was predominantly observed below the mucosal layer. Furthermore, based on various specific and immunohistochemical studies as well as the electronmicroscopic findings, the patients were pathologically diagnosed to have gallbladder metastasis of renal cell carcinoma. PMID- 11871828 TI - Elevated serum pro-mMP2 levels in patients with advanced lung cancer are not suitable as a prognostic marker. AB - Using the one-step sandwich enzyme immunoassay for pro-MMP2, we investigated whether the serum pro-MMP2 levels could be used as predictors of the development and extension of lung cancer. The study included 52 lung cancer patients and 26 nonmalignant thoracic disease patients. Serum samples were collected before clinical treatment. The serum pro-MMP2 levels were found to be elevated in patients with lung cancer (1006.3+/-37.4 ng/ml) in comparison with nonmalignant controls (767.7+/-41.6 ng/ml) (P = 0.0002). In addition, stage IIIb lung cancer patients (1167.2+/-124.2 ng/ml) had significantly higher pro-MMP2 levels compared with stage I patients (936.0+/-44.8 ng/ml, P = 0.0431). The serum pro-MMP2 levels were similar in the samples between the pathological subtypes. As a result, the serum pro-MMP2 levels may serve as a marker that can be used as a diagnostic indicator of higher stages in lung cancer. There was no difference in the prognosis between the group with normal pro-MMP2 levels (<850 ng/ml) and the group of elevated pro-MMP2 levels (>850 ng/ml), thus indicating that serum analyses of the pro-MMP2 levels were of limited value in the prognosis of lung cancer. PMID- 11871829 TI - Methylprednisolone acetate versus oral prednisolone in moderately active ulcerative colitis. AB - BACKGROUND: Patients with active ulcerative colitis are treated with corticosteroids. We compared the efficacy and safety of intramuscular depot preparation of methylprednisolone acetate with oral prednisolone in the treatment of moderately active ulcerative colitis. DESIGN: Open labeled, randomized, prospective, four-month study. METHODS: 40 patients with moderately active ulcerative colitis (activity index 150-220) were randomized into two groups. Group A (n=21) received methylprednisolone acetate (80 mg intramuscularly once weekly for 6 weeks). Group B (n=19) received oral prednisolone (40 mg/day) in a 'tailing-off' regimen. In addition, patients in both the groups received sulfasalazine. Patients were followed up at 1, 2, 3, 4, 8, 12 and 16 weeks. The primary measure of therapeutic response was activity index. An index of <150 was considered as clinical remission. Secondary efficacy was assessed by subjective evaluation of acceptability of treatment by the patient. RESULTS: After one week of treatment, the decrease in mean activity index was significantly more with oral prednisolone (p<0.05), and five 5 patients (23.8%) in Group A and 12 (63.2%) in Group B were in clinical remission (p<0.05). However, after 2 weeks and beyond, the mean activity index and the number of patients with clinical remission were comparable in the two treatment groups. CONCLUSIONS: Methylprednisolone acetate as a depot preparation and oral prednisolone are equally effective in inducing remission in patients with moderately active ulcerative colitis. Though symptomatic improvement is quicker with oral prednisolone, the remission rate with the two drugs was similar after 2 weeks of treatment. PMID- 11871830 TI - Role of travel as a risk factor for hepatitis E virus infection in a disease endemic area. AB - BACKGROUND: We undertook epidemiologic and laboratory studies during an epidemic of acute hepatitis in Sindri town, in District Dhanbad, Bihar in 1998. METHODS: A sample survey covering 201 randomly selected houses in the town was conducted during the epidemic, and records of patients admitted to the only large hospital in this town were reviewed. We also tested serum and stool specimens from some of the affected persons for hepatitis E virus (HEV) RNA and IgM anti-HEV antibodies. RESULTS: Of the 1088 persons residing in the surveyed houses, 82 (7.54%) had developed acute hepatitis during the outbreak. Attack rate was higher among male residents than among female residents (71/604 vs. 11/484; 11.75% vs. 2.27%; relative risk [RR] 5.17 [95% confidence interval 2.77-9.65]; p<10(-6)) and was the highest in the 10-29 year age group. Hospital admission data showed similar age and gender distribution. Disease occurrence had no relation with source of drinking water (handpump 7.56% vs. municipal tap 7.53%; p=ns), or with habit of boiling (RR 1.10 [0.61-1.98]; p=ns) or filtering (RR 0.59 [0.33-1.06]; p=ns) water before drinking. Jaundice occurred more frequently among persons who had traveled outside Sindri town during the last two months than among those who had not (26.4% vs. 4.7%; RR 5.67 [3.81-8.43]; p<10(-6)); this risk persisted after correction for age (Mantel-Haenszel weighted OR 6.74 [4.12-11.01]; p<10(-6)). Men traveled more frequently than women and were more often affected. In multivariate analysis, travel and male gender were the only two independent risk factors. Data from a hospital in a neighboring large city, Dhanbad, suggested that there was an outbreak of hepatitis in that city too at the same time. Seventy-three of the 1088 study subjects had history of jaundice in the past; disease attack rate among these persons (9.6%) was similar to that among those without such history (7.5%; RR 1.31 [0.49-2.98]; p=ns). Of the 13 sera tested, 10 were positive for IgM anti-HEV. HEV RNA was detected in 9 of the 12 stool specimens and 10 of the 13 sera tested. CONCLUSIONS: The hepatitis epidemic in Sindri was caused by HEV and had several features resembling those of previous HEV epidemics. However, the occurrence of hepatitis E showed a strong relationship with history of travel, a finding not hitherto described. PMID- 11871831 TI - Helicobacter pylori infection delays ulcer healing in patients operated on for perforated duodenal ulcer. AB - BACKGROUND: A majority of duodenal ulcers are associated with Helicobacter pylori infection; eradication of the infection improves ulcer healing and reduces the ulcer recurrence rate. However, the frequency of H. pylori infection in patients with perforated duodenal ulcer is not clearly established. We studied the frequency of H. pylori infection in patients with perforated duodenal ulcer and its impact on their clinical presentation. METHODS: All patients presenting with perforated duodenal ulcer underwent emergency laparotomy and simple omental patch repair. Postoperatively they received standard antibiotics for 1-2 weeks along with ranitidine; ranitidine alone was continued thereafter till an endoscopy 4-6 weeks later. Positive rapid urease test along with identification of H. pylori on histology was taken as evidence of H. pylori infection. Patients who received anti-H. pylori therapy or nonsteroidal anti-inflammatory drugs (NSAIDs) postoperatively and/or proton pump inhibitors or antibiotics, during 4 weeks preceding the endoscopy, were excluded. RESULTS: 30 patients (27 men; mean [SD] age 32.9 [9.7 years]) presenting during the period June 1999 to October 2000 were studied. Upper gastrointestinal endoscopy was done 10.9 (6.3) weeks after surgery. Seventeen (56.6%) patients were infected with H. pylori; this group had significantly more men (17/17 versus 10/13 among uninfected) and fewer NSAID users (2/17 vs. 7/13). Median duration of epigastric pain before presentation was 18 weeks in the H. pylori-infected group as compared to one week in the non infected group (p<0.001). Significantly more patients continued to have epigastric pain after surgery in the infected group (7/17 vs. 0/13). At endoscopy, active duodenal ulcer was present in 13 of 17 patients with evidence of H. pylori infection and none of the noninfected patients (p<0.001). Age, sex, duration between surgery and endoscopy, NSAID use, smoking and maintenance ranitidine use had no impact on ulcer healing after the surgery. CONCLUSIONS: In patients operated on for perforated duodenal ulcer, H. pylori infection was the only significant factor responsible for persistence of ulcer after surgery. We advocate H. pylori eradication therapy in patients operated on for perforated duodenal ulcer. PMID- 11871832 TI - Should we eradicate Helicobacter pylori to improve gastric histology? PMID- 11871833 TI - Metronidazole resistance in Helicobacter pylori: magnitude, mechanism and implications for India. PMID- 11871834 TI - Gastric schwannoma presenting as gastric polyp with gastrointestinal bleeding. AB - Benign neurogenic tumors of stomach are the commonest of all nonepithelial tumors of the stomach, but solitary gastric schwannomas are rare. We report a 58-year old man with gastric schwannoma presenting as fundal polyp and upper gastrointestinal bleeding. Endoscopic polypectomy was done. PMID- 11871835 TI - Massive bleeding from duodenal diverticulum. AB - Duodenal diverticula are commonly located in the second part of the duodenum and are usually asymptomatic. We report a 45-year-old man with massive bleeding from a duodenal diverticulum located beyond the second part of the duodenum. The diverticulum was excised, and the patient. has remained asymptomatic over the next 12 months. PMID- 11871836 TI - Pancreatitis in enteric fever. AB - Pancreatitis in enteric fever is rare. We report two patients with enteric fever, one due to Salmonella typhi infection and other due to S. paratyphi, who on investigation were found to have pancreatitis. Both patients recovered uneventfully. PMID- 11871837 TI - Nephrotic syndrome cured by removal of malignant gastric stromal tumor. AB - Malignant tumors have been associated with the development of nephrotic syndrome. We report a 73-year-old man with nephrotic syndrome who had a malignant stromal gastric tumor. Curative resection of the sarcoma resulted in remission of the nephrotic syndrome. PMID- 11871838 TI - Cystic variant of embryonal sarcoma of liver. AB - Embryonal sarcoma of the liver is an unusual tumor. The cystic form is rare and can mimic hydatid disease. We present a case that was mistakenly treated as a hydatid cyst for 3 months. Surgery was successful in removing the mass. PMID- 11871840 TI - Severe acute pancreatitis in acute hepatitis E. AB - We report an 18-year-old boy with severe acute pancreatitis developing during acute hepatitis E and complicated by sepsis and acute renal failure. The patient recovered on supportive management. PMID- 11871839 TI - Percutaneous expulsion of hydatid liver cyst following sclerotherapy. AB - A 20-year-old woman underwent alcohol sclerotherapy for a symptomatic liver cyst, initially diagnosed as simple liver cyst. Five months later, she presented with expulsion of the cyst from the puncture site. The cyst was proved to be due to hydatid disease. Spontaneous expulsion is a rare complication of sclerotherapy. PMID- 11871841 TI - Concept of mucosal plug in the etiology of duodenal ulcer. PMID- 11871843 TI - Epidemiology of hepatitis A in India. PMID- 11871844 TI - Total aganglionosis coli. PMID- 11871842 TI - Comparison of effects of cholecystokinin and erythromycin on bile chemistry and gallstone formation in aged guinea pigs. AB - BACKGROUND: There has been considerable interest in gall bladder motility in recent years. We compared the effects of cholecystokinin (CCK) and erythromycin on bile chemistry and gallstone formation in aged guinea pigs. METHODS: Two groups of guinea pigs (1-mo and 3-y old; n=40 each) were studied. Each group was divided into four subgroups of 10 animals each; one subgroup received lithogenic diet, one each received CCK or erythromycin daily in addition to lithogenic diet for 4 weeks, and one received normal diet. After 4 weeks, the presence of gallstones or sludge was recorded and bile composition including concentrations of bile acid, cholesterol, lecithin and protein concentrations was studied. RESULTS: No gallstones were observed in the 1-mo-old animals. In the 3-year-old animals, 9 of 10 guinea pigs on lithogenic diet and 4 of 10 in each treatment subgroup and the normal diet subgroup developed gallstones. CCK and erythromycin had similar effects on bile chemistry and stone formation. CONCLUSIONS: Aging increases the formation of gallstones in guinea pigs. Erythromycin is as effective as CCK in reducing gallstone formation by improving gall bladder motility. PMID- 11871845 TI - Crohn's disease of stomach. PMID- 11871846 TI - Pre and post eradication gastric inflammation in Helicobacter pylori-associated duodenal ulcer. AB - BACKGROUND: Distribution and nature of gastritis are major determinants of clinical outcome of H. pylori infection. The gastric inflammatory changes associated with this infection in developing countries have not been systematically studied. AIMS: To evaluate the inflammatory changes in gastric antrum and corpus in patients with duodenal ulcer and H. pylori infection, before and after H. pylori eradication therapy. METHODS: Histology and H. pylori density were studied in gastric biopsies obtained from 53 consecutive patients with active duodenal ulcer and H. pylori infection. Biopsies were obtained before and 4 weeks after H. pylori eradication therapy, from the anterior and posterior walls of the antrum and corpus, and were evaluated according to the Sydney system. RESULTS: In the pre-H. py/ori eradication antral biopsies, chronic gastritis, active gastritis, atrophy, intestinal metaplasia (IM) and lymphoid follicles / aggregates were seen in 53 (100%), 49 (92%), 11 (21%), 7 (13%) and 28 (53%) patients, respectively. In the corresponding biopsies from gastric corpus, these changes were seen in 49 (92%), 23 (43%), 2 (4%), 2 (4%) and 8 (15%), respectively. All changes except IM were significantly more frequent and of higher grade in the antrum. The grade of chronic gastritis was significantly higher in antrum than corpus; the frequency of gastritis in the antrum and corpus was similar (100% vs. 92%). H. pylori density was also higher in the antrum and correlated well with the grades of chronic gastritis and activity at both sites. Eradication of H. pylori was achieved in 39 patients (74%), and led to significant decrease in gastritis; no change was seen in patients who did not eradicate the organism. CONCLUSIONS: Antral-predominant chronic gastritis and activity are present in more than 90% of patients with H. pylori infection associated with duodenal ulcer, and the grade of gastritis correlates with the density of the organism. Eradication therapy results in improvement of both chronic gastritis and activity. PMID- 11871847 TI - The psychosocial work environment and health--what do we know and where should we go? PMID- 11871848 TI - Effects of ergonomic intervention in work with video display units. AB - OBJECTIVES: This study evaluated the effect of an intensive ergonomic approach and education on workstation changes and musculoskeletal disorders among workers who used a video display unit (VDU). METHODS: A randomized controlled design was used. The subjects (N=124) were allocated into three groups (intensive ergonomics, ergonomic education, reference) using stratified random sampling. The evaluation involved questionnaires, a diary of discomfort, measurements of workload, and an ergonomic rating of the workstations. The assessments were made 2 weeks before the intervention and after 2 and 10 months of follow-up. RESULTS: The intensive and training groups showed less musculoskeletal discomfort than the reference group after 2 months of follow-up. Positive effects on discomfort were seen primarily for the shoulder, neck, and upper back areas. No significant differences were found for the strain levels or prevalence of pain. After the intervention the ergonomic level was distinctly higher in the intensive ergonomic group than in the education or reference group. CONCLUSIONS: Both the intensive ergonomics approach and education in ergonomics help reduce discomfort in VDU work. In attempts to improve the physical ergonomics of VDU workstations, the best result will be achieved with cooperative planning in which both workers and practitioners are actively involved. PMID- 11871849 TI - Physical workload and the risk of severe knee osteoarthritis. AB - OBJECTIVES: This study investigated the impact of physical workload on the risk of severe knee osteoarthritis (OA) leading to knee arthroplasty. METHODS: In this case-referent study, the cases were 55- to 75-year-old men (N=55) and women (N=226) who had undergone their first knee arthroplasty operation for primary knee OA in the Kuopio University Hospital in 1992-1993. The referents (N=524) were from the same source population and were matched with the cases for age and gender. Information on explanatory variables was obtained by a computer-assisted telephone interview. Exposure was assessed up to 49 years of age. RESULTS: After adjustment for body mass index, knee injury and leisure-time physical activity, an increased relative risk of knee OA was found for a history of high physical workload. The odds ratio (OR) was 1.53 [95% confidence interval (95% CI) 0.42 5.56] for the men and 2.03 (95% CI 1.03-3.99) for the women as compared with those with low physical workload. With respect to generic risk factors, climbing already at a medium level of exposure was associated with an increased risk of knee OA among the men (OR 3.06, 95% CI 1.25-7.46) and kneeling and squatting was a risk factor for both genders. Of the different trades, agriculture, forestry, fishing, transportation, and traffic showed the highest risks of knee OA. CONCLUSIONS: The results of this study agree with the hypotheses that heavy physical loading increases the risk of knee OA and that cumulative physical stress has a deleterious effect on the knee joint. PMID- 11871850 TI - Prevalence of self-reported hypersensitivity to electric or magnetic fields in a population-based questionnaire survey. AB - OBJECTIVES: The prevalence of medically unexplained symptoms attributed to exposure to electromagnetic fields is still largely unknown. Previous studies have investigated reported hypersensitivity to electricity in selected groups recruited from workplaces or outpatient clinics. The aim of this study was to estimate the prevalence of self-reported hypersensitivity to electric or magnetic fields in the general population and to describe characteristics of the group reporting such hypersensitivity with regard to demographics, other complaints, hypersensitivities, and traditional allergies. METHODS: A cross-sectional questionnaire survey was conducted in 1997 among 15,000 men and women between 19 and 80 years of age in Stockholm County. The response rate was 73%. RESULTS: One and a half percent of the respondents reported hypersensitivity to electric or magnetic fields. Prevalence was highest among women and in the 60- to 69-year age group. The hypersensitive group reported all symptoms, allergies, and other types of hypersensitivities included in the survey (as well as being disturbed by various factors in the home) to a significantly greater extent than the rest of the respondents. No specific symptom profile set off the hypersensitive group from the rest of the respondents. CONCLUSIONS: The results should be interpreted with caution. But they suggest that there is widespread concern among the general population about risks to health posed by electric and magnetic fields. More research is warranted to explore ill health among people reporting hypersensitivity to electric or magnetic fields. PMID- 11871852 TI - Exploration of asthma risk by occupation--extended analysis of an incidence study of the Finnish population. AB - OBJECTIVES: The objective of the study was to determine asthma risks at the most detailed level of occupational classification in a previously described nationwide follow-up study that included the entire employed workforce of Finland. METHODS: In Finland, persons with clinically verified persistent asthma are registered for medication reimbursement within the national health insurance scheme. Data were combined from three national registers, and all 25- to 59-year old employed Finns were followed for asthma incidence in 1986-1998. Altogether 49,575 cases were detected. A log-linear model was used to estimate the relative risks of asthma for 275 nonadministrative occupations in comparison with administrative work (33 occupations). RESULTS: A significantly increased risk was found for either men or women in 125 occupations. For the men, the risk was highest among bakers, laundry workers, shoemakers and repairers, tanners, fell mongers and pelt dressers, and metal plating and coating workers. For the women, the risk was highest among shoemakers and repairers, railway and station personnel, jewelry engravers, engineroom crew, molders, round-timber workers. and bakers. CONCLUSIONS: The results suggest that the work-related excess of asthma incidence is much more widely spread across the labor force than has been previously thought. A great number of occupations deserves to be targeted for in depth studies focusing on the determinants of asthma excess and on possibilities for better asthma control among asthmatics working in these occupations. The large work-relatedness of asthma incidence should also raise public health interest because of the economic losses incurred and the potential for prevention. PMID- 11871851 TI - Occupational exposure to magnetic fields in case-referent studies of neurodegenerative diseases. AB - OBJECTIVES: Case-referent studies of Alzheimer's disease, amyotrophic lateral sclerosis, and Parkinson's disease were conducted to explore the relationship between these neurodegenerative diseases and occupational exposure to magnetic fields. Three methods of exposure assessment were used for the comparison, and the consistency of findings between these approaches was evaluated. METHODS: Separate case-referent sets were formed from among recorded deaths of males in the state of Colorado for the years 1987 through 1996. The following three methods of exposure assessment were used: a dichotomous grouping of electrical versus nonelectrical occupations, a three-tiered grouping of potential magnetic field exposure based on a combination of job title and industry, and categories of exposure based on the means of the magnetic fields estimated from a job exposure matrix. RESULTS: A positive association was observed for Parkinson's disease with all the methods of magnetic-field exposure assessment, the odds ratio (OR) for the highest category in the job-exposure matrix being 1.50 [95% confidence interval (95% CI) 1.02-2.19]. Amyotrophic lateral sclerosis was associated with a history of electrical occupations (OR 2.30, 95% CI 1.29-4.09) but not with magnetic-field exposure as estimated by the job-exposure matrix. No consistent associations with magnetic fields were observed for Alzheimer's disease. CONCLUSIONS: This study provides some support for an association between occupational magnetic-field exposure and Parkinson's disease, but the findings are novel and require replication. Associations with the other neurodegenerative diseases were inconsistent and dependent on the method of exposure assessment. PMID- 11871853 TI - Adverse reproduction outcomes among employees working in biomedical research laboratories. AB - OBJECTIVES: The aim of the study was to investigate reproductive outcomes such as birthweight, preterm births, and postterrm births among women working in research laboratories while pregnant. METHODS: Female university personnel were identified from a source cohort of Swedish laboratory employees, and the database was linked to the medical birth register. The first births of the women were included in the analysis, 249 pregnancies among the women with laboratory work and 613 pregnancies among the women without laboratory tasks. Information about exposure to various laboratory agents was obtained from a previous questionnaire investigation at the research group level according to a specific definition. The ponderal index and ratio between observed and expected birthweights were calculated. Logistic regression models were used for analyses of dichotomous outcomes (preterm, postterrm and birthweight). RESULTS: Exposure to laboratory work with solvents was associated with an increased risk of preterm births, the estimated odds ratio (OR) being 3.4 (1.0 < 95% confidence interval < 11.9). An association with work with bacteria was also observed for postterm births (OR 2.7, 1.0 < or = 95% confidence interval < 7.4). CONCLUSIONS: There was a slightly elevated risk for some reproductive outcomes among the women working with certain laboratory tasks, specifically for preterm and postterm births in relation to work with solvents and bacteria. PMID- 11871854 TI - Alternative for estimating the burden of lung cancer from occupational exposures- some calculations based on data from Swedish men. AB - OBJECTIVES: This study attempts to demonstrate a calculation of the occupational lung cancer burden using economically active men in Sweden as an example. METHODS: Estimates were calculated using Swedish register data on occupation in 1970, lung cancer incidence in 1971-1989, smoking frequencies in 1963, and the formula I = RI0F + I0(1-F), where I is the overall incidence, R is the relative risk associated with a factor (here smoking), F is the fraction of persons at risk (smokers), and I0 is the incidence among those not at risk (nonsmokers). RESULTS: Farmers, gardeners, forestry workers, and fishermen had the lowest lung cancer risk (42.1 per 100,000 person-years) and a smoking frequency of 44.7%. Their I0 was 12.6 or 8.4 per 100,000 person-years, taking R for smoking as 6 or 10, respectively. From these I0 estimates, the expected rates for white- and blue collar workers (smoking frequencies 52.7 and 57.7%, respectively) were 45.8 and 49.1 per 100,000 person-years, as compared with the 22% and 57% higher observed rates, respectively. Weighing these excesses proportionally according to the sizes of the three occupational categories gave, respectively for R equal to 6 and 10, occupation-related excesses of 39% and 32% and population-attributable risks of 28% and 24%. CONCLUSIONS: About one-fourth of the lung cancers that occur among economically active Swedish men seem to have been related to occupation. This figure agrees with estimates made by other methods in Nordic countries. Due to interaction, the population-attributable risk from smoking is still high, 73% and 83% at relative risk values of 6 and 10, respectively. PMID- 11871855 TI - Shift work and age as interactive predictors of body mass index among offshore workers. AB - OBJECTIVES: This study investigated shift pattern (day shifts versus day-night rotation) and its interactions with age, and with years of shiftwork exposure, as predictors of body mass index (BMI). METHODS: Survey data were collected from offshore personnel working day shifts (N=787) or day-night shifts (N=787); information was obtained about shift pattern and years of shiftwork exposure, height, weight, demographic factors, and smoking habits. Hierarchical multiple regression was used to test a model in which BMI was predicted by additive and interactive effects of shift pattern, age, and exposure years with control for confounding variables. RESULTS: In a multivariate analysis (controlling for job type, education and smoking), BMI was predicted by the main effects of age and years of shiftwork exposure. Shift pattern was not significant as a main effect, but it interacted significantly with the curvilinear age term and with the linear and curvilinear components of shiftwork exposure. In the day shift group, age but not exposure predicted BMI; the opposite was true of the day-night shift group. The increase in BMI with an increase in age and exposure years was steeper for the day-night shift group than for the day shift group. CONCLUSIONS: The significant interaction effects found in this study were consistent with the view that continued exposure to day-night shift work gives rise to increases in BMI, over and above the normative effects of ageing on BMI shown by day-shift workers. PMID- 11871856 TI - Protooncogene Bcl-2 induces apoptosis in several cell lines. AB - Using a recombinant vaccinia virus expressing protooncogene Bcl-2, we demonstrate opposite effects of the expressed Bcl-2 in two cell lines: apoptosis induction in BSC-40 cells and apoptosis prevention in HeLa G cells. The apparent molecular weight of the expressed Bcl-2, its amounts and its effects on the mitochondrial membrane potential are comparable in both cell lines, suggesting that the consequences of Bcl-2 expression depend on the cellular environment. To further support these findings we demonstrate the pro-apoptotic effect of the expressed Bcl-2 in several other cell lines. PMID- 11871857 TI - The regulatory region of Prague C v-Src inhibits the activity of the Schmidt Ruppin A v-Src kinase domain. AB - Existing variants of the oncogene v-src differ in their transforming potential as well as in the range of their hosts. We compared the protein kinase activities of two Prague C v-Src variants (PRC and H19), reported to be of low oncogenic potential (Plachy et al., 1995), with the highly oncogenic Schmidt-Ruppin A v-Src (SRA). We employed in vitro kinase assays of affinity-purified proteins expressed in rabbit reticulocyte lysate and in S. cerevisiae. In both systems used, the specific kinase activity of the Prague C v-Src kinases amounted to only ca 20% of the activity of SRA. This positions the PRC Src close to activated c-Src, despite the lack of the regulatory C-terminal tail in PRC. We constructed chimeras between PRC and SRA v-Src and tested them for specific kinase activity in S. cerevisiae. Remarkably, the regulatory N-terminal part of PRC, when fused to the SRA-derived kinase domain, lowered the chimeras' PK activity to ca 20%, suggesting that it is the regulatory part of PRC that is responsible for its low phosphotransferase activity. PMID- 11871858 TI - Microdissection techniques for cancer analysis. AB - One difficulty in studying molecular changes of tumours has been the inability to isolate DNA and RNA from a homogeneous cell population. The combination of several new technologies should help overcome these hurdles. Microdissection is a technique for rapid and easy procurement of a pure cellular subpopulation away from its complex tissue milieu. Laser-assisted microdissection has recently been identified as a quick, simple and effective method by which microdissection of complex tissue specimens can be routinely performed for molecular analysis. With the advent of laser microdissection, cDNA libraries can be developed from pure cells obtained directly from stained neoplastic tissue, and microarrays of thousands of genes can now be used to examine gene expression in microdissected tumour tissue samples. This review will concentrate on the application of different microdissection techniques in the area of cancer research. PMID- 11871859 TI - Genomic organization of the mouse src gene. Sequencing of src introns revealed a new chromosome 2 microsatellite marker. AB - Ten introns interrupting the coding sequence of the mouse src protooncogene were sequenced (in total 11260 bp) and their general characteristics compared with the homologous genes in human and chicken. While the study of genome organization of the src gene was performed only in the inbred mouse strain BALB/cHeA (Mus musculus domesticus), one special region in the intron 5 was also sequenced in additional mouse strains (M. musculus musculus and M. spretus), because the discovered CA and GT repeat array differences could serve as a new polymorphic marker in the chromosome No. 2 and help elucidate some evolutionary relations between mouse strains. PMID- 11871860 TI - New monoclonal antibody to human apolipoprotein J. PMID- 11871861 TI - Chromosomal mapping of HCaRG, a novel hypertension-related, calcium-regulated gene. AB - We recently identified a novel gene that is negatively regulated by extracellular calcium concentration with higher levels of transcripts in hypertensive animals (SHR). We named this gene HCaRG (Hypertension-related, Calcium-Regulated Gene). In this work we report the chromosomal localization of the HCaRG gene among different species. We identified a BglII RFLP between BN.lx and SHR rats. We then analysed the strain distribution pattern of this RFLP in 31 RIS, originating from BN.lx and SHR rats, and compared it to the segregation of 475 markers localized in the rat genetic map. Hcarg localizes to the rat chromosome 7 between the markers Mit3 and Mit4. This region is homologous to human chromosome 8q21-24. We identified three clones in Genbank that contain the sequence of HCaRG. It was therefore possible to narrow down the localization of human HCaRG to chromosome 8q24.3. Furthermore, a suggestive localization of mouse Hcarg based on conservation of linkage between human and mouse is on chromosome 15. We previously identified a putative calcium-binding motif (EF-Hand) and a nuclear receptor-binding domain (LxxLL) in the rat sequence of the HCaRG protein. Sequence comparison between five different species showed that these domains are highly conserved. Furthermore, a search of ESTs in Genbank for homologous sequences showed that HCaRG is expressed only in eukaryotes, particularly in mammals. PMID- 11871862 TI - Breast cancer prevention: present and future. AB - Increased risk of breast cancer may result from modifiable factors such as endogenous hormone levels, obesity, HRT, and non-lactation, or non-modifiable factors such as genetic susceptibility or increasing age. Those factors that are easiest to modify may have a limited impact on the totality of breast cancer. The Gail model, based on known factors may be useful for estimating life-time risk in some individuals. Tamoxifen prevention still remains contentious. In the NSABP-P1 study, there was a 49% reduction in risk of breast cancer in women given tamoxifen but in the Italian and Royal Marsden trials, no effect on breast cancer incidence was detected, possibly because of the different case-mix in these studies. Raloxifene, tested in the MORE trial reduced the incidence of breast cancer by 65%. The effect was restricted to ER positive tumours: no reduction in ER negative cancers was seen. Life-style factors such as diet, obesity, exercise, and age of first full term pregnancy and number of pregnancies have a mild to moderate impact on risk and so may have little effect on the incidence of breast cancer. Reduction of alcohol intake could lead to a modest reduction in the risk of breast cancer but possibly adversely affect other diseases. So far, studies of retinoids have not shown a benefit in terms of breast cancer risk reduction. Fat reduction and GnRH analogues reduce mammographic density but have not yet been shown to affect risk. PMID- 11871863 TI - Nipple discharge: current diagnostic and therapeutic approaches. AB - Nipple discharge is a complex diagnostic challenge for the clinician. A variety of diseases (such as intraductal papillomas, mammary duct ectasia, breast cancer, pituitary adenomas, breast abscesses/infections, etc.) can manifest as nipple discharge. The importance of nipple discharge for both the patient and the physician is the possible association of this condition with an underlying carcinoma. With heightened public awareness of breast cancer, an increasing number of women are asking their health care providers about nipple discharge. A detailed clinical evaluation is invaluable to determine the pathophysiology, assess the risk of malignancy, and plan treatment of the patient with nipple discharge. A combination of diagnostic tests, including mammography, breast ultrasonography, and possibly galactography can help the clinician to establish the diagnosis and plan proper management. Depending on the underlying breast pathology, a central or single lactiferous duct excision is the procedure of choice. Breast carcinoma associated with nipple discharge should be treated by either a modified radical mastectomy of breast-conservation therapy (i.e. duct lobular segmentectomy with adequate, free margins [ideally>1cm], levels I and II axillary lymph node dissection, followed by breast irradiation). PMID- 11871864 TI - Chemotherapy in patients with metastatic or relapsed germ-cell tumours. AB - The optimal treatment in patients with poor prognosis germ-cell tumours (GCT), according to the International Germ Cell Cancer Collaborative Group (IGCCCG) classification, and in patients with refractory or relapsed disease after cisplatin-based chemotherapy is controversial. As the majority of patients will suffer systemic relapses, chemotherapy is the mainstay of treatment. However, the question of whether or not to use conventional-dose or high-dose chemotherapy (HDCT) in these patients arises. Prognostic factors have recently been recognised to aid in this decision. However, reliable data on chemotherapy as primary treatment in poor prognosis patients and as the first-salvage attempt in patients with relapsed or refractory GCT are lacking. This report reviews the recent developments in first-line and salvage HDCT strategies and discusses the role of predictive factors for treatment outcome. PMID- 11871865 TI - Defining a future role for radiogenic therapy. AB - The goal of cancer therapy is to eliminate the cancer and/or to arrest further growth while decreasing normal tissue toxicity, i.e. to increase the therapeutic ratio. This review focuses on a group of therapeutics that are either (1) directly stimulated by radiation to produce either directly or indirectly cytotoxic agents (i.e. genes under the control of a radiation inducible promoter that produce a cytotoxic protein or an enzyme that converts a prodrug to an active form, respectively); (2) auger-electron emitting radiolabelled oligonucleotides, antibodies, nucleotide analogues, or other small molecules that are internalized; (3) radiation inducible genes that produce a ligand or transporter (or the like) which then can be targeted by cytotoxic agents (e.g. radiolabelled substance). We have termed this group of therapeutics radiogenic therapy. PMID- 11871866 TI - BRCA-1 and BRCA-2 mutations as prognostic factors in clinical practice and genetic counselling. AB - Women in general have a 10% risk of developing breast cancer and a 2-3% chance of ovarian cancer in their life-times. Mutations in BRCA-1 and BRCA-2 are present in only a small portion (5-10%) of all breast cancers. Carriers of mutations in these genes have a greater risk of cancer, especially before menopause in the case of BRCA-1 carriers. In addition, their risk of contralateral breast cancer is significantly higher than for the general population (4.2-53% vs. 2%). The grade of contralateral tumours in these patients is more aggressive. BRCA-2 hereditary breast cancer seems more heterogeneous than the BRCA-1 phenotype, and not clearly different from sporadic forms. However, since 20-30% of carriers of BRCA mutations never develop breast or ovarian cancer, there must be other 'risk modifiers'. Survival is better for carriers of hereditary ovarian cancer. Patients with these mutations are referred for genetic counselling, a complex process which includes: an informative dialogue between the proband and the geneticist, drawing up a family history, informed consent, evaluation of risk, genetic testing and possible involvement of healthy family members. PMID- 11871867 TI - Pruritus in cancer patients. AB - Pruritus is an uncommon symptom in cancer patients but can often be difficult to treat. The pathogenesis is complex and is not fully understood although there is evidence of involvement of a number of mediators from which treatment options are developing. In cancer patients pruritus may be directly related to the cancer, indirectly related (e.g. cholestasis) or associated with treatment. It is not always possible to treat the underlying cause of the pruritus in these patients, or desirable to stop treatments that may contribute and in these cases we must address the pruritus itself. The treatment of pruritus is a developing area, and helpful research is slowly emerging. The evidence base is not extensive but some high quality studies exist. This short paper briefly discusses the pathogenesis, causes, effects and treatment options for pruritus in cancer patients. PMID- 11871868 TI - Fickle hexadienes. Manipulating the relative energies of chairlike and boatlike transition structures for the cope rearrangement. AB - We report calculations on various hexadienes that can assume both chairlike and boatlike conformations, yet turn out (theoretically) to have transition structures for boatlike Cope rearrangement that are equal to or lower in energy than those of alternative chairlike structures. Pathways connecting boatlike and chairlike transition states in these systems also allow for unusual and facile isomerization pathways of certain strained alkenes. PMID- 11871869 TI - A density-functional study of the mechanism for the diastereoselective epoxidation of chiral allylic alcohols by the titanium peroxy complexes. AB - The epoxidation of three stereolabeled methyl-substituted chiral allylic alcohols with (1,2)A and/or (1,3)A allylic strain, namely 3-methylbut-3-en-2-ol (1a), pent 3-en-2-ol (1b), and 3-methylpent-3-en-2-ol (1c), have been studied by the density functional theory method, B3LYP/6-31+G(d,p). For each substrate we calculated the two prereaction complexes with Ti(OH)(4)/MeOOH (the oxidant model for Ti(O-i Pr)(4)/t-BuOOH), their threo and erythro transition states for oxygen transfer, and the corresponding product complexes. For substrate 1a, the erythro transition state is 0.91 kcal/mol of lower energy than the threo one; for substrates 1b and 1c, the threo compared to the erythro transition states are by 1.05 and 0.21 kcal/mol more favorable, respectively. The threo/erythro product ratios have been estimated from the computed free energies for the competing threo and erythro transition states 3a-c in CH(2)Cl(2) solution to be 12:88 (1a), 92:8 (1b), and 77:23 (1c), which are in good accordance with the experimental values 22:78 (1a), 91:9 (1b), and 83:17 (1c). The diastereoselectivity of this diastereoselective oxyfunctionalization is rationalized in terms of the competition between (1,3)A and (1,2)A strain and the electronic advantage for the spiro transition state. In addition, solvent effects are also play a role for the diastereoselectivity at the same time. PMID- 11871870 TI - Templated conversion of a crown ether-containing macrobicycle into [2]rotaxanes. AB - A crown ether-containing macrobicycle was used as the wheel component in a templated synthesis of a [2]rotaxane with an acetal-containing axle. The molecular structures of the macrobicycle and the [2]rotaxane were characterized by NMR spectroscopy and X-ray crystallography. The chloride-binding ability of the macrobicycle, either free in solution or when it is part of a [2]rotaxane, is quite weak as determined by NMR titration experiments. A second analogous [2]rotaxane, with a longer axle, was synthesized, and its solvent-dependent co conformation was characterized by 2D NMR spectroscopy. The position of the wheel along the axle can be controlled by the solvent polarity, however, attempts to use metal cations such as Na(+), K(+), Ba(2+), and Ag(+) to switch the wheel position in polar solvents were unsuccessful. PMID- 11871871 TI - The azine bridge as a conjugation stopper: an NMR spectroscopic study of electron delocalization in acetophenone azines. AB - Dipole parallel-alignment of organic molecular crystals of azines has been achieved with a design that was based on the hypothesis that the azine bridge is a conjugation stopper. This hypothesis has now been tested in detail, and (1)H and (13)C NMR spectroscopic data of symmetric and asymmetric acetophenone azines are presented in support of this design concept. Previous structural, ab initio, and electrochemical studies have shown that the azine bridge largely inhibits through-conjugation in molecules with the general structure DPhC(Me) [double bond] N [bond] N [double bond] C(Me)PhA, where D is a donor group and A is an acceptor group. NMR spectroscopy is an excellent tool to probe the degree of conjugation through the azine bridge. The NMR results reported here for nine symmetrical and 18 asymmetrical azines show in a compelling fashion that the hypothesis holds. Varying the donor group does not change the chemical shifts of the aromatic hydrogen and carbon atoms on the acceptor-substituted phenyl ring. Likewise, varying the acceptor group does not change the chemical shifts of the atoms in the donor-substituted phenyl ring. PMID- 11871873 TI - Aromaticity of the Bergman, Myers-Saito, Schmittel, and directly related cyclizations of enediynes. AB - Aromaticity criteria (magnetic susceptibility exaltations, nucleus independent chemical shifts (NICS), and aromatic stabilization energy (ASE) evaluations) for enediyne and enyne-allene cyclizations evaluated at (UBS)-BLYP/6-31G* all agree that the degrees of cyclic electron delocalization of the benzenoid systems formed by the Bergman (3) and Myers-Saito reactions (5) are comparable to benzene. The reaction enthalpy differences between the parent cyclizations and their benzannelated analogues are not entirely due to disparities in gained ASE during the reactions. The alternative formation of fulvene biradicals is not accompanied by favorable aromatic effects. PMID- 11871874 TI - Tautomerism in the solid state and in solution of a series of 6-aminofulvene-1 aldimines. AB - To study systems able to sustain intramolecular proton-transfer, we have prepared a series of six aminofulvene aldimines including several labeled with (15)N and (2)H. These compounds show coupling constants through the hydrogen bond, (1h)J((15)N- (1)H) and (2h)J((15)N-(15)N). The position of the tautomeric equilibria, i.e., on what nitrogen atom is the proton, was determined in the solid state and in solution. The crystal structure of N[[5 [(phenylamino)methylene]-1,3-cyclopentadien-1-yl]methylene]pyrrole-1-amine (3) has been determined by X-ray analysis. In solution, both N-H and C-H tautomers were observed and their structures assigned by NMR spectroscopy. Particularly useful is the value of the (1)J((15)N-(1)H) coupling constant. PMID- 11871872 TI - Enantioselective synthesis of N(alpha)-Fmoc protected (2S,3R)-3-phenylpipecolic acid. A constrained phenylalanine analogue suitably protected for solid-phase peptide synthesis. AB - Reported herein is the first enantioselective preparation of (2S,3R)-3 phenylpipecolic acid as a conformationally constrained phenylalanine analogue bearing N(alpha)-protection suitable for solid-phase peptide synthesis. Stereochemistries at both the 2- and 3-positions are derived inductively from a single chiral center provided by the commercially available Evans chiral auxiliary, (4S)-4-benzyl-1,3-oxazolidin-2-one. By constraining phi and chi(1) torsion angles, this novel amino acid analogue can serve as a useful tool for the induction of defined geometry in phenylalanine-containing peptides. PMID- 11871877 TI - Solvolysis of 1,1-dimethyl-4-alkenyl chlorides: evidence for pi-participation. AB - Tertiary 1,1-dimethyl-4-alkenyl chloride (1) solvolyzes with significantly reduced secondary beta-deuterium kinetic isotope effect (substrate with two trideuteromethyl groups) and has a lower entropy and enthalpy of activation than the referent saturated analogue 4 (k(H)/k(D) = 1.30 +/- 0.03 vs k(H)/k(D) = 1.79 +/- 0.01; Delta Delta H(++) = -9 kJ mol(-1), Delta Delta S(++) = -36 J mol(-1) K( 1), in 80% v/v aqueous ethanol), indicating participation of the double bond in the rate-determining step. Transition structure 1-TS computed at the MP2(fc)/6 31G(d) level of theory revealed that the reaction proceeds through a late transition state with considerably pronounced double bond participation and a substantially cleaved C-Cl bond. The doubly unsaturated compound 3 (1,1-dimethyl 4,8-alkadienyl chloride) solvolyzes with further reduction of the isotope effect, and a drastically lower entropy of activation (k(H)/k(D) = 1.14 +/- 0.01; DeltaS(++) = -152 +/- 12 J mol(-1) K(-1), in 80% v/v aqueous ethanol), suggesting that the solvolysis of 3 proceeds by way of extended pi-participation, i.e., the assistance of both double bonds in the rate-determining step. PMID- 11871875 TI - 5-Dethia-5-oxacephams: toward correlation of absolute configuration and chiroptical properties. AB - The relationship between chiroptical properties of differently substituted 5 dethia-5-oxacephams and their respective molecular structures was investigated. The amide chromophore of the beta-lactam unit in these compounds was found to be nonplanar with a shallow pyramidal configuration at the nitrogen atom. Due to the nonplanarity, the beta-lactam system becomes inherently dissymmetric, which is supported by a high magnitude of the n --> pi* CD band. It was also found that the helicity of the lactam moiety in investigated oxacephams is controlled by the absolute configuration at the C(6) carbon atom. On this basis, a helicity rule correlating a positive (negative) sign of the n right arrow pi Cotton effect with a negative (positive) O [double bond] C [bond] N [bond] C(6) torsional angle for policyclic beta-lactam derivatives possessing a nonplanar amide chromophore was formulated. PMID- 11871878 TI - Asymmetric syntheses of new functionalized beta-amino alcohols via diastereoselective addition of organometallic reagents onto oxazolidines. AB - Diastereoselective reactions between (S)-phenylglycinol-derived oxazolidines and two unsaturated organolithium reagents afforded chiral beta-amino alcohols having vinyl and alkynylsilane moieties. When the same reactions were performed in the presence of titanium isopropoxide, a dramatic change of regioselectivity was observed, from the alpha- to the gamma-position, thus producing beta-amino alcohols with allyl or allenylsilane functions. A rationalization of the observed diastereoselectivity was suggested for each case. PMID- 11871879 TI - Unexpected hydrodeiodo Sonogashira-Heck-Casser coupling reaction of 2,2' diiodobiphenyls with acetylenes. AB - 2,2'-Diiodobiphenyl-4,4'-dicarboxylic acid dimethyl ester (3) undergoes either a ring-closure reaction with phenylacetylene to give 4 or hydrodeiodo phenylethynylation to give 5 under the catalytic conditions of Pd(OAc)(2)/CuI/phosphine in amines. In these reactions, the amine and the phosphine ligands play important roles in controlling the reactivity. Among the ligands we used, tris(o-tolyl)phosphine is the best ligand for hydrodeiodo phenylethynylation, while the bidentate phosphine ligand retards both of the reactions. On the basis of our results, we propose that 5 is formed through a fast hydrodeiodination, followed by a Sonogashira phenylethynylation. The results of the deuterium labeling experiments show that proton exchange between the acetylenic proton and the alkyl protons of amine occurs effectively under the reaction conditions. In addition, the hydrogen that replaces the iodide in the hydrodeiodination process arises mainly from the acetylenic proton. PMID- 11871876 TI - One-pot two-step enzymatic coupling of pyrimidine bases to 2-deoxy-D-ribose-5 phosphate. A new strategy in the synthesis of stable isotope labeled deoxynucleosides. AB - The enzymatic synthesis of thymidine from 2-deoxy-D-ribose-5-phosphate is achieved, in a one-pot two-step reaction using phosphoribomutase (PRM) and commercially available thymidine phosphorylase (TP). In the first step the sugar 5-phosphate is enzymatically rearranged to alpha-2-deoxy-D-ribose-1-phosphate. Highly active PRM is easily obtained from genetically modified overproducing E. coli cells (12,000 units/84 mg protein) and is used without further purification. In the second step thymine is coupled to the sugar-1-phosphate. The thermodynamically unfavorable equilibrium is shifted to the product by addition of MnCl(2) to precipitate inorganic phosphate. In this way the overall yield of the beta-anomeric pure nucleoside increases from 14 to 60%. In contrast to uracil, cytosine is not accepted by TP as a substrate. Therefore, 2'-deoxy cytidine is obtained by functional group transformations of the enzymatically prepared 2'-deoxy-uridine. The method has been demonstrated by the synthesis of [2',5'-(13)C(2)]- and [1',2',5'-(13)C(3)]thymidine as well as [1',2',5' (13)C(3)]2'-deoxyuridine and [3',4'-(13)C(2)]2'-deoxycytidine. In addition the nucleoside bases thymine and uracil are tetralabeled at the (1,3-(15)N(2),2,4 (13)C(2))-atomic positions. All compounds are prepared without any scrambling or dilution of the labeled material and are thus obtained with a very high isotope enrichment (96-99%). In combination with the methods that have been developed earlier it is concluded that each of the (13)C- and (15)N-positions and combination of positions of the pyrimidine deoxynucleosides can be efficiently labeled starting from commercially available and highly (13)C- or (15)N-enriched formaldehyde, acetaldehyde, acetic acid, potassium cyanide, methylamine hydrochloride, and ammonia. PMID- 11871881 TI - Influence of intermolecular hydrogen bonds on the tautomerism of pyridine derivatives. AB - The effect of the dimerization, by hydrogen-bond (HB) complexation, on the tautomerism of 2-hydroxypyridine and a series 2-aminopyridines has been carried using ab initio methods. The results obtained for 2-hydroxypyridine fit satisfactorily with the experimental data and show that the 2-pyridone/2-pyridone homodimer complex is the most stable. For 2-aminopyridines, the effect of the substituent on the amino group has been investigated. For the monomers studied, the most favorable tautomer is the 2H; however, with electronegative substituents, the 1H/1H homodimers are more stable than the corresponding 2H/2H ones. The atom in molecule methodology has been used to characterize the HBs formed. Exponential relationships have been found between the electron density and its laplacian at the HB critical point vs the HB distance. PMID- 11871880 TI - Iron(III) [bond] Salen complexes as enzyme models: mechanistic study of oxo(salen)iron complexes oxygenation of organic sulfides. AB - The oxidation of a series of para-substituted phenyl methyl sulfides was carried out with several oxo(salen)iron (salen = N,N'-bis(salicylidine)ethylenediaminato) complexes in acetonitrile. The oxo complex [O=Fe(IV)(salen)](*+), generated from an iron(III) [bond] salen complex and iodosylbenzene, effectively oxidizes the organic sulfides to the corresponding sulfoxides. The formation of [O [double bond] Fe(IV)(salen)](*+) as the active oxidant is supported by resonance Raman studies. The kinetic data indicate that the reaction is first-order in the oxidant and fractional-order with respect to sulfide. The observed saturation kinetics of the reaction and spectral data indicate that the substrate binds to the oxidant before the rate-controlling step. The rate constant (k) values for the product formation step determined using Michaelis-Menten kinetics correlate well with Hammett sigma constants, giving reaction constant (rho) values in the range of -0.65 to -1.54 for different oxo(salen)iron complexes. The log k values observed in the oxidation of each aryl methyl sulfide by substituted oxo(salen)iron complexes also correlate with Hammett sigma constants, giving positive rho values. The substituent effect, UV-vis absorption, and EPR spectral studies indicate oxygen atom transfer from the oxidant to the substrate in the rate-determining step. PMID- 11871882 TI - Tributylstannyl radical-catalyzed reaction of 1,2,3-selenadiazoles with olefins or dienes. AB - It was found that the reaction of 1,2,3-selenadiazoles derived from cyclic ketones with olefins or dienes was markedly promoted by a catalytic amount of tributylstannyl radical, which was generated in situ from tributylstannyl hydride or allyltributylstannane and AIBN, to give the corresponding dihydroselenophenes in moderate to good yields. In contrast, when 1,2,3-selenadiazoles prepared from linear and aromatic ketones were used as substrates, the same reaction did not take place, and alkynes were formed as the sole product. PMID- 11871883 TI - Computational characterization of sulfur-cxygen-bonded sulfuranyl radicals derived from alkyl- and (carboxyalkyl)thiopropionic acids: evidence for sigma type radicals. AB - Sulfide radical cations are known to stabilize via sulfur-oxygen bond formation with carboxylic acid functions. However, structural information on the resulting sulfuranyl radicals has so far only been provided by ESR spectroscopy for more stable, aromatic-substituted species, while the rather short-lived aliphatic sulfuranyl radicals have largely been characterized by time-resolved UV spectroscopy only. Therefore, we have obtained theoretical structural information on S-O-bonded sulfuranyl radicals from three model compounds, tert-butyl 2 (methylthio)peroxybenzoate, 3-methylthiopropionic acid, and 3,3'-thiodipropionic acid, using density functional theory, semiempirical, and molecular mechanics methods. All S-O-bonded species exist predominantly as three-electron-bonded sigma-radicals with an estimated heterolytic bond dissociation energy of the S therefore O bond on the order of 23-27 kcal mol(-1). Characteristic optical absorption bands, vibrational frequencies, and hyperfine coupling tensors are evaluated to facilitate the identification of such radicals by time-resolved UV, IR, and ESR spectroscopy. PMID- 11871885 TI - Synthesis and membrane activity of a bis(metacyclophane)bolaamphiphile. AB - The synthesis of macrocyclic tetraesters from 5-substituted isophthalic acids and 1,8-octane diol gave macrocylic metacyclophanes with the axially symmetric positions differentially protected. Dimer bis(metacyclophane)bolaamphiphiles proved to be extremely insoluble, but one derivative was shown to have significant membrane activity. Very high continuous conductance and the formation of discrete single-ion channels were both observed, depending on how the compound was incorporated into the bilayer membrane. PMID- 11871884 TI - Serine and threonine beta-lactones: a new class of hepatitis A virus 3C cysteine proteinase inhibitors. AB - Hepatitis A virus (HAV) 3C enzyme is a cysteine proteinase essential for viral replication and infectivity and represents a target for the development of antiviral drugs. A number of serine and threonine beta-lactones were synthesized and tested against HAV 3C proteinase. The D-N-Cbz-serine beta-lactone 5a displays competitive reversible inhibition with a K(i) value of 1.50 x 10(-6) M. Its enantiomer, L-N-Cbz-serine beta-lactone 5b is an irreversible inactivator with k(inact) = 0.70 min(-1), K(Iota) = 1.84 x 10(-4) M and k(inact)/K(Iota) = 3800 M( 1) min(-1). Mass spectrometry and HMQC NMR studies using (13)C-labeled 5b show that inactivation of the enzyme occurs by nucleophilic attack of the cysteine thiol (Cys-172) at the beta-position of the oxetanone ring. Although the N-Cbz serine beta-lactones 5a and 5b display potent inhibition, other related analogues with an N-Cbz side chain, such as the five-membered ring homoserine gamma lactones 14a and 14b, the four-membered ring beta-lactam 33, 2-methylene oxetane 34, cyclobutanone 36, and 3-azetidinone 39, fail to give significant inhibition of HAV 3C proteinase, thus demonstrating the importance of the beta-lactone ring for binding. PMID- 11871886 TI - Total synthesis of nor-1,6-germacradien-5-ols. AB - The first total synthesis of (+/-)-nor-1,6-germacradien-5-ols is described. The synthetic route involves the RCM methodology for the ring formation and a selective 1,2 addition of MeLi to cyclodecenone. By altering the order of the last synthetic steps, TBSO-protected (+/-)-(1Z,6E)-nor-1,6-germacradien-5-ols (+/ )-(5S*,8R*)-16 and -(+/-)-(5S*,8S*)-16 were obtained. The synthetic strategy via cyclodecenone offers the possibility of preparing different analogues of the title compounds through addition of other nucleophiles. Moreover, nor-germacrene D could be accessed from the target molecule by methylenation of its carbonyl moiety. (+/-)-nor-1,6-Germacradien-5-ol [(+/-)-(1E,5S*,6E,8S*)-2] was synthesized in eight steps from isovaleric acid. The 10-membered ring was formed by RCM, and the tertiary alcohol moiety was introduced in the last step via a highly diastereoselective addition of MeLi to (+/-)-(1E,6E)-1,6-cyclodecen-5-one (+/-) E,E-5. Addition of MeLi to cyclodecenone (+/-)-Z,E-5 also occurred with complete selectivity to provide (+/-)-(1Z,5S*,6E,8S*)-2. A slightly different synthetic pathway was also explored, in which the order of the final synthetic steps was switched: the enone formation and the addition of MeLi were conducted prior to the cyclization. When the hydroxy group was protected as a TBS ether, the newly formed olefin had exclusively Z configuration. Thus, TBSO-protected (+/-)-(1Z,6E) nor-1,6-germacradien-5-ols (+/-)-16 were obtained as a 1:1 (5S*,8S*)/(5R*,8S*) mixture. The NMR spectra of these two diastereomers confirmed the relative stereochemistry of natural (-)-1,6-germacradien-5-ol (1) at C5 and C8. PMID- 11871887 TI - Aigialomycins A-E, new resorcylic macrolides from the marine mangrove fungus Aigialus parvus. AB - Aigialomycins A-E (2-6), new 14-membered resorcylic macrolides, were isolated together with a known hypothemycin (1) from the mangrove fungus, Aigialus parvus BCC 5311. Structures of these compounds, including absolute configuration, were elucidated by spectroscopic methods, chemical conversions, and X-ray crystallographic analysis. Hypothemycin and aigialomycin D (5) exhibited in vitro antimalarial activity with IC(50) values of 2.2 and 6.6 microg/mL, respectively, while other analogues were inactive. Cytotoxicities of these compounds were also evaluated. PMID- 11871888 TI - New catalysts for the base-promoted isomerization of epoxides to allylic alcohols. Broadened scope and near-perfect asymmetric induction. AB - Optically active (1S,3R,4R)-3-[N-(trans-2,5-dialkyl)pyrrolidinyl]methyl-2 azabicyclo-[2.2.1]heptanes were evaluated as catalysts for the enantioselective beta-elimination of meso-epoxides. The (2R,5R)-dimethylpyrrolidinyl-substituted catalyst 4 exhibited exceptionally high enantioselectivity and reactivity, and several substrates were rearranged with enantioselectivities of 98-99% ee. In addition, the use of 4 allowed the first successful, true catalytic rearrangement of the difficult substrates cyclopentene oxide (81%, 96% ee) and (Z)-4-octene oxide (80%, 91% ee). PMID- 11871889 TI - Microwave specific Wolff rearrangement of alpha-diazoketones and its relevance to the nonthermal and thermal effect. AB - alpha-Diazoketones possess high electric dipole moments, as a consequence of the dipolar nature of the diazocarbonyl functional group. The vectorial analysis, theoretical calculations (PM3 and ab initio), and literature reports based on experimental and theoretical calculations reveal a higher dipole moment for the Z configuration of the diazo functional group. Microwave irradiation of alpha diazoketone (1a-m) (Figure 1) promotes Wolff rearrangement specifically via the Z configuration in excellent yields. The dielectric properties of the solvent govern the course of the microwave rearrangement. 3-Diazocamphor (1m) on microwave irradiation in benzylamine exhibits nonthermal effects to furnish exclusively the Wolff rearrangement product (4m), equivalent to its photochemical behavior. In the presence of an aqueous medium, through solvent heating predominates, leading to the formation of a tricyclic ketone (5) as the principal product, arising from an intramolecular C-H insertion. This behavior is similar to its known thermal and transition metal catalyzed reactivity pattern. PMID- 11871891 TI - Syntheses, derivatives, solubility, and interfacial properties of 2-methyl-2 polyfluoroalkenyloxymethyl-1,3-propanediols: potential building blocks for syntheses of amphiphatic macromolecules. AB - 2-Hydroxymethyl-2-methyl-1,3-propanediol (A) was reacted with (Me(3)Si)(2)NH and toluenesulfonyl chloride (TsCl) to give mainly CH(3)C(CH(2)OSiMe(3))(3) (1), and CH(3)C(CH(2)OTs)(3) (2), respectively. With allyl bromide, the products were CH(3)C(CH(2)OCH(2)CH[double bond]CH(2))(2)(CH(2)OH) (3) and CH(3)C(CH(2)OCH(2)CH[double bond]CH(2))(CH(2)OH)(2) x H(2)O (4). The reactions of 4 with perfluoroalkyl iodides (R(f)I) were catalyzed by Cu(I)Cl to form 2-methyl 2-polyfluoroalkenyloxymethyl-1,3-propanediols: (R(f)CH=CHCH(2)OCH(2))C(Me)(CH(2)OH)(2) [R(f) = C(4)F(9) (5), C(8)F(17) (6), and (CF(2)CF(2))(4)OCF(CF(3))(2) (7)]. Reduction of 5 and 6 with hydrogen gave two new 2-methyl-2-polyfluoroalkyloxymethyl-1,3-propanediols, 8 and 9. The sodium salt of 9 was reacted with allyl bromide or acetyl chloride to form (C(8)F(17)CH(2)CH(2)CH(2)OCH(2))C(Me)(CH(2)OX)(CH(2)OH)(2) [where X = CH(2)CH=CH(2) (10) or C(O)CH(3) (12)] and (C(8)F(17)CH(2)CH(2)CH(2)OCH(2))C(Me)(CH(2)OX)(2) [where X = CH(2)CH[double bond]CH(2) (11) or C(O)CH(3) (13)]. Reaction of tolenesulfonyl chloride with 7 gave the monotosylate, 14, as the sole product. With 4-trifluoromethylbenzyl bromide, the sodium salt of 4 gave (4 CF(3)C(6)H(4)CH(2)OCH(2))C(Me)(CH(2)CH[double bond]CH(2))(CH(2)OH) x H(2)O (15). The compounds were characterized by NMR ((1)H, (13)C, (19)F, (29)Si), GC-MS, and high-resolution MS or elemental analyses. UV evidence was obtained for partitioning of 9, 12, 14, and 15 between perfluorodecalin and n-octanol. The test compounds acted as surfactants by facilitating the solubility of phenol and Si(CH[double bond]CH(2))(4) in perfluorodecalin. The single-crystal X-ray structure of 8 was also obtained. It crystallized in the monoclinic space group P2(1)/c, and unit cell dimensions were a = 24.966(2) A (alpha = 90), b = 6.1371(6) A (beta = 100.730(2)), and c = 10.5669(10) A (gamma = 90). PMID- 11871890 TI - Efficient synthesis of the A-ring phosphine oxide building block useful for 1 alpha,25-dihydroxy vitamin D(3) and analogues. AB - The 1 alpha-hydroxy A-ring phosphine oxide 1, a useful building block for vitamin D analogues, was synthesized from (S)-carvone in nine synthetic operations and a single chromatographic purification in 25% overall yield. The synthesis features two novel efficient synthetic transformations: the Criegee rearrangement of alpha methoxy hydroperoxyacetate 10 in methanol to obtain directly the desired secondary 3 beta-alcohol 11 and the highly chemo- and stereoselective isomerization of dieneoxide ester (E)-7 to the 1 alpha-allylic alcohol with an exocyclic double bond (E)-8. Further insight into the selectivity control of the latter rearrangement was obtained from the reactions of (Z)-epimeric substrates. The new synthetic approach leading to the 1 alpha-hydroxy epimers complements our previously reported synthesis of the corresponding 1 beta-epimers, thus producing all stereoisomers of these versatile building blocks efficiently from carvone. PMID- 11871892 TI - A new method for the preparation of 2-thio substituted furans by methylsulfanylation of gamma-dithiane carbonyl compounds. AB - Several related methods for the preparation of differentially substituted 2 thiofurans are described. The general procedure involves the formation of a thionium ion from a gamma-dithianyl substituted carbonyl compound followed by cyclization of this reactive intermediate onto the tethered carbonyl group. Two methods for thionium ion generation were explored. One of these involved an acid catalyzed reaction of beta-ketenedithioacetals, prepared from the condensation of 2,2-bis(methylsulfanyl)acetaldehyde with a variety of ketones. Cyclization followed by loss of methane thiol gave 2-thiofurans 17, 18 and 23, 24 in 70-90% yield. Attempts to prepare 5-heteroatom substituted 2-thiofurans from the corresponding beta-ketenedithioacetal amides or esters were unsuccessful, leading to 1,2-thio rearranged products. A more successful route involved the reaction of beta-acetoxy-gamma-thianyl carbonyl compounds with dimethyl(methylthio)sulfonium tetrafluoroborate (DMTSF). Treatment of the dithiane with this reagent resulted in the smooth generation of a thionium ion. Cyclization followed by loss of acetic acid afforded thiofurans 17, 18, 23, 47-49, 51, and 61-64 in 40-100% yield. The N-butenyl substituted thioamido furan furnished a rearranged hexahydropyrroloquinolin-2-one in high yield when heated at 110 degrees C. PMID- 11871893 TI - Total synthesis of the ethyl ester of the major urinary metabolite of prostaglandin E(2). AB - The preparation of the ethyl ester of the major urinary metabolite of prostaglandin E(2) 3 is described. The key step is the kinetic opening of the TBS protected bicyclic ketone 7 with thiophenol. PMID- 11871894 TI - Substituent effects on the acidity of weak acids. 1. Bicyclo[2.2.2]octane-1 carboxylic acids and bicyclo[1.1.1]pentane-1-carboxylic acids. AB - The acidities of 3- and 4-substituted bicyclooctane-1-carboxylic acids and 3 substituted bicyclo[1.1.1]pentane-1-carboxylic acids have been calculated at the MP2/6-311++G** theoretical level. There is good agreement between the calculated and observed gas-phase acidities. The acidities of the 4-substituted bicyclooctane acids were found to be linearly dependent on the C-X bond dipoles, as expected from a field effect. The substituents had a negligible effect on the electron density at C1. The difference in acidity between 4 chlorobicyclo[2.2.2]octane-1-carboxylic acid and the parent acid (6.2 kcal/mol) is reproduced by the Kirkwood-Westheimer treatment of substituent effects on acidity, but only if the bicyclooctane ring is given an effective dielectric constant of unity. The acidities of the 3-substituted bicyclooctane acids are linearly related to the corresponding 4-substituted acids with a slope of 0.9. The acidities of the 3-substituted bicyclo[1.1.1]pentane-1-carboxylic acids are linearly related to the C-X bond dipoles for this ring system (which are different than those for the bicyclooctanes), and they are also linearly related to the acidity of the 4-substituted bicyclo[2.2.2]octanecarboxylic acids with a slope of 1.34. The larger slope is due to the smaller bridgehead-bridgehead distance in the bicyclopentane ring than in bicyclo[2.2.2]octane. PMID- 11871895 TI - Influence of the beta-alkoxy group on the diastereoselectivity of Aldol reactions of tetrahydro-4H-thiopyran-4-one with 4-Alkoxytetrahydro-2H-thiopyran-3 carboxaldehydes. AB - The diastereoselectivity of the aldol reaction of tetrahydro-4H-thiopyran-4-one (3) with 1,4-dioxa-8-thiaspiro[4.5]decane-6-carboxaldehyde (9a) under a variety of conditions is examined. Under optimized conditions, three of the four possible diastereomers from this aldol reaction can be obtained selectively (3-16:1). Reactions of 9a with the Li, B, Mg(II), and Ti(IV) enolates of 3 and with the corresponding trimethylsilyl enol ether 4b in the presence of BF(3) x OEt(2), SnCl(4), or TiCl(4) as promoters gave the Felkin adducts exclusively (>95%) as mixtures of syn (11a) and anti (12a) diastereomers. Use of the "amine-free" Li enolate of 3 gave 12a with a much higher diastereoselectivity (9:1) and yield (70%) than that obtained using the lithium diisopropylamide-generated Li enolate of 3 (2-3:1; 15-40%). The TiCl(4)-promoted reaction of 4b with 9a gave 11a with excellent selectivity (16:1). In contrast, the MgBr(2) x OEt(2)-promoted reaction of 4b with 9a gave the anti-Felkin adducts exclusively as a 3:1 mixture of syn (13a)/anti (14a) diastereomers. Similar aldol reactions of 3 with the cis and trans isomers of 4-(methoxy)methoxytetrahydro-2H-thiopyran-3-carboxaldehyde (9b and 9c) were examined to probe the influence of the ketal protecting group in 9a on the observed aldol diastereoselectivity. The results are rationalized by applying Evans' stereochemical model for merged 1,2- and 1,3-asymmetric induction (non-chelation), with the exception of the MgBr(2) x OEt(2)-promoted reactions of 4b with 9a, 9b, and 9c, which are accommodated by assuming chelation control. Comparison of the reactions of 9a, 9b, and 9c suggests that the ketal group in 9a uniquely allows high levels of either Felkin or anti-Felkin selectivity to be achieved. PMID- 11871896 TI - Hydride-transfer domino rearrangement of glycine-containing dioxa-azawurtzitane. AB - The novel synthetic method for dioxa-azawurtzitanes to selectively cap amino groups in amino acids or peptides is described. Mixing the CH(3)CN solution of cis,cis-1,3,5-triformyl-1,3,5-trimethylcyclohexane (2) with the aqueous solution of the equimolar amounts of glycine and NaHCO(3) yields glycine-containing dioxa azawurtzitane 7-Na. Dioxa-azawurtzitane 7-Na almost quantitatively isomerizes to lactone-imine 9-Na through the hydride-transfer rearrangement in CH(3)CN/H(2)O. Lactone-imine 9-Na also isomerizes to lactam-aldehyde 12-Na in DMSO. PMID- 11871897 TI - Synthesis and NMR studies of (13)C-labeled vitamin D metabolites. AB - Isotope-labeled drug molecules may be useful for probing by NMR spectroscopy the conformation of ligand associated with biological hosts such as membranes and proteins. Triple-labeled [7,9,19-(13)C(3)]-vitamin D(3) (56), its 25-hydroxylated and 1 alpha,25-dihydroxylated metabolites (58 and 68, respectively), and other labeled materials have been synthesized via coupling of [9-(13)C]-Grundmann's ketone 39 or its protected 25-hydroxy derivative 43 with labeled A ring enyne fragments 25 or 26. The labeled CD-ring fragment 39 was prepared by a sequence involving Grignard addition of [(13)C]-methylmagnesium iodide to Grundmann's enone 28, oxidative cleavage, functional group modifications leading to seco iodide 38, and finally a kinetic enolate S(N)2 cycloalkylation. The C-7,19 double labeling of the A-ring enyne was achieved by the Corey-Fuchs/Wittig processes on keto aldehyde 11. By employing these labeled fragments in the Wilson-Mazur route, the C-7,9,19 triple-(13)C-labeled metabolites 56, 58, and 68 as well as other (13)C-labeled metabolites have been prepared. In an initial NMR investigation of one of the labeled metabolites prepared in this study, namely [7,9,19-(13)C(3)] 25-hydroxyvitamin D(3) (58), the three (13)C-labeled carbons of the otherwise water insoluble steroid could be clearly detected by (13)C NMR analysis at 0.1 mM in a mixture of CD(3)OD/D(2)O (60/40) or in aqueous dimethylcyclodextrin solution and at 2 mM in 20 mM sodium dodecyl sulfate (SDS) aqueous micellar solution. In the SDS micellar solution, a double half-filter NOESY experiment revealed that the distance between the H(19Z) and H(7) protons is significantly shorter than that of the corresponding distance calculated from the solid state (X-ray) structure of the free ligand. The NMR data in micelles reveals that 58 exists essentially completely in the alpha-conformer with the 3 beta-hydroxyl equatorially oriented, just as in the solid state. The shortened distance (H(19Z))-H(7)) in micellar solutions as compared to that in the solid state is most easily rationalized on the basis that the 5(10)-torsion angle in 58 is decreased in micellar solutions as compared to that in the solid state. PMID- 11871899 TI - Efficient ruthenium-catalyzed aerobic oxidation of alcohols using a biomimetic coupled catalytic system. AB - Efficient aerobic oxidation of alcohols was developed via a biomimetic catalytic system. The principle for this aerobic oxidation is reminiscent of biological oxidation of alcohols via the respiratory chain and involves selective electron/proton transfer. A substrate-selective catalyst (ruthenium complex 1) dehydrogenates the alcohol, and the hydrogens abstracted are transferred to an electron-rich quinone (4b). The hydroquinone thus formed is continuously reoxidized by air with the aid of an oxygen-activating Co[bond]salen type complex (6). Most alcohols are oxidized to ketones in high yield and selectivity within 1 2 h, and the catalytic system tolerates a wide range of O(2) concentrations without being deactivated. Compared to other ruthenium-catalyzed aerobic oxidations this new catalytic system has high turnover frequency (TOF). PMID- 11871898 TI - Absolute stereochemistry of amphidinolide E. AB - The absolute configurations at eight chiral centers in amphidinolide E (1), a cytotoxic 19-membered macrolide isolated from a marine dinoflagellate Amphidinium sp., were determined to be 2R, 7R, 8R, 13S, 16S, 17R, 18R, and 19R on the basis of detailed analysis of NMR data and by chemical means. PMID- 11871900 TI - Conformational studies by dynamic NMR. 86. Structure, stereodynamics, and cryogenic enantioseparation of the stereolabile isomers of o-dinaphthylphenyl derivatives. AB - Static and dynamic stereochemistry of the hydrocarbon comprising a phenyl ring bearing two alpha-naphthyl substituents in the ortho positions, i.e., 1,2-di-(4 methyl-naphth-1-yl)-benzene 1, has been studied by a combination of variable temperature NMR, cryogenic HPLC, and MM calculations. Whereas in solution both syn (meso) and anti (chiral) forms were observed and the corresponding interconversion barrier was determined (Delta G(++) = 19.5 kcal mol(-1)), only the diastereoisomer anti was found to be present in the crystalline state (X-ray diffraction). When the molecule is rendered asymmetric by introduction of a nitro group in the phenyl ring as in 1,2-di-(4-methyl-naphth-1-yl)-4-nitrobenzene 2, the chiral syn and anti diastereoisomers are simultaneously present both in solution and in the solid state, albeit in different proportions. Cryogenic chromatography on a HPLC chiral stationary phase at 20 degrees C allowed the stereolabile diastereoisomers and the corresponding enantiomers to be separated. PMID- 11871901 TI - Development of versatile cis- and trans-dicarbon-substituted chiral cyclopropane units: synthesis of (1S,2R)- and (1R,2R)-2-aminomethyl-1-(1H-imidazol-4 yl)cyclopropanes and their enantiomers as conformationally restricted analogues of histamine. AB - The cyclopropane ring can be used effectively in restricting the conformation of biologically active compounds to improve activity and also to investigate bioactive conformations. We designed (1S,2R)- and (1R,2R)-2-aminomethyl-1-(1H imidazol-4-yl)cyclopropanes (1 and 2, respectively) and their enantiomers (ent-1 and ent-2) as conformationally restricted analogues of histamine. The four types of chiral cyclopropanes bearing two differentially functionalized carbon substituents in a cis or trans relationship on a cyclopropane ring, (1S,2R)-2 (tert-butyldiphenylsilyloxy)methyl-1-formylcyclopropane (7) and (1R,2R)-2-(tert butyldiphenylsilyloxy)methyl-1-formylcyclopropane (8) and their enantiomers (ent 7 and ent-8), were developed as the key intermediates for synthesizing 1, 2, ent 1, and ent-2. The reaction between (R)-epichlorohydrin [(R)-12] and phenylsulfonylacetonitrile (13a) in the presence of NaOEt in EtOH followed by treatment with acid gave the chiral cyclopropane lactone 11a with 98% ee in 82% yield. Compound 11a was converted into both the cis- and trans-chiral cyclopropane units 7 and 8, respectively, via reductive desulfonylation with Mg/MeOH as the key step. The corresponding enantiomers, the cis-substituted ent-7 and the trans-substituted ent-8, were also prepared starting from (S) epichlorohydrin [(S)-12]. The four conformationally restricted target histamine analogues 1, 2, ent-1, and ent-2 were successfully synthesized from 7, 8, ent-7, and ent-8, respectively. The chiral cyclopropane units 7, 8, ent-7, and ent-8 should be useful as versatile intermediates for synthesizing various compounds having an asymmetric cyclopropane structure. PMID- 11871902 TI - Straightforward synthesis of enantiomerically pure (3S,4R)- and (3R,4S)-3,4 isopropylidenedioxypyrroline 1-oxide, precursors of functionalized cis-dihydroxy azaheterocycles, by a novel "one-pot" procedure. AB - The enantiomerically pure nitrone 3, a valuable precursor of mono- and bicyclic azaheterocycles, has been synthesized in 57% yield by a novel "one-pot" process starting from lactol 1, in turn readily available from D-arabinose. The same process, consisting of reaction with a O-silyl-protected hydroxylamine followed by mesylation in pyridine, furnished ent-3 in 55% yield when applied to L arabinose. PMID- 11871903 TI - Nickel-catalyzed coupling of aryl iodides with aromatic aldehydes: chemoselective synthesis of ketones. AB - Aryl iodides (ArI) couple with aryl aldehydes (Ar'CHO) in the presence of Ni(dppe)Br(2) and Zn to give the corresponding biaryl ketones (ArCOAr'). The use of a bidentate phosphine complex is critical to the success of this catalytic reaction. The reaction provides a new procedure for the synthesis of various functionalized biaryl ketones. PMID- 11871904 TI - Synthesis of enantiopure omega-functionalized C15 alpha-amino carboxylates. AB - An efficient route for the synthesis of enantiopure omega-hydroxy, omega-carboxy, omega-oxo, and omega-amino alpha-amino acids and bis-alpha-amino acids was developed. The synthesis of omega-trityloxy delta,epsilon-unsaturated alpha-amino acids was based on the Wittig reaction of methyl (2S)-2-[bis(tert butoxycarbonyl)amino]-5-oxopentanoate with omega-trityloxy alkylidene triphenylphosphoranes. After hydrogenation, the omega-hydroxy alpha-amino acid was used as starting material for the synthesis of other omega-functionalized alpha-amino acids. The length of the side chain of alpha-amino acids or bis-alpha amino acids depends on the starting alkanediol or dibromide used to prepare the phosphoranes. PMID- 11871905 TI - Squaric acid ester-based total synthesis of echinochrome A. AB - The total synthesis of echinochrome A is described. Both key intermediates 5 and 8 were efficiently prepared from diisopropyl squarate 7. Nucleophilic addition of aryllithium 8 to 5, followed by thermal ring-expansion/cyclization of the 1,2 adduct 4, furnished hydroquinone 3. Oxidation and full deprotection of 3 gave the title compound. PMID- 11871907 TI - Discovery of C(60)O(3) isomer having C(3)(nu) symmetry. AB - A hitherto undetected type of C(60)O(3) isomer was found in the reaction solution of C(60) with m-chloroperoxybenzoic acid by means of a chromatographic technique using two different columns. Both electronic spectroscopy and atmospheric pressure chemical ionization (APCI) mass-spectroscopy examinations show its C(3)(nu) symmetry, in which three oxygen atoms are added onto one benzenoid ring of C(60). PMID- 11871906 TI - Efficient synthesis of medium-sized cyclic ether diamines. PMID- 11871908 TI - Facile synthesis of N-aryl pyrroles via Cu(II)-mediated cross coupling of electron deficient pyrroles and arylboronic acids. AB - N-Arylation of electron-deficient pyrroles has been achieved by cross-coupling with arylboronic acids at room temperature in the presence of stoichiometric amounts of copper(II) acetate. The generality of this reaction has been established with variously substituted pyrroles as well as boronic acids. A key intermediate in the synthesis of a matrix metalloprotease inhibitor has been achieved using this methodology. PMID- 11871909 TI - Novel C-C bond cleavage under mild, neutral conditions: conversion of electron deficient aryl alkyl ketones to aryl carboxylic esters. AB - A novel, unique way to cleave the carbon-carbon bond in aryl alkyl ketones under mild, neutral conditions is described. Treatment of aryl alkyl ketones in a refluxing mixture of N,N-dimethylformamide dimethyl acetal and methanol for 16 h provided aryl carboxylic esters. The scope and limitations of the reaction are discussed. Useful yields of the reaction can be obtained with electron-deficient aryl groups, and the yields are higher when the alkyl group is larger than a methyl group. Studies toward elucidation of the reaction mechanism led to a proposed mechanism that is consistent with all the observations. PMID- 11871910 TI - Stereoselective additions of chiral (E)-crotylsilanes to thionium ions: asymmetric synthesis of homoallylic thioethers. AB - Stereochemically well-defined homoallylic thioethers 3 are synthesized via Lewis acid promoted condensation reaction between chiral organosilane reagents 2 and in situ generated thionium ions. The stereochemical course of the reaction is consistent with earlier reports concerning crotylsilations of oxonium ions. PMID- 11871911 TI - Aromatic amination/imination approach to chiral benzimidazoles. AB - The powerful Buchwald-Hartwig amination was utilized for the construction of the benzimidazole nucleus with the substituted nitrogen atom bearing a chiral substituent. A successive amination/imination was followed by an acid-catalyzed ring closure step to give the benzimidazole ring. The products were deprotonated and acylated at the C2 position and could be alkylated on nitrogen to give chiral benzimidazolium salts. PMID- 11871912 TI - Practical synthesis of optically active amino alcohols via asymmetric transfer hydrogenation of functionalized aromatic ketones. AB - 2-Substituted acetophenones such as 2-cyano-, 2-azido-, or 2-nitroacetophenones were effectively reduced with a mixture of HCOOH/N(C(2)H(5))(3) containing a chiral Ru(II) catalyst, RuCl[(S,S)-N-(p-toluenesulfonyl)-1,2 diphenylethylenediamine](p-cymene), giving the corresponding optically active alcohols, which can be converted to optically active amino alcohols with excellent ee's. Similarly, the reaction of 2-benzoylacetophenone with the same Ru catalyst gave a quantitative yield of the corresponding optically active 1,3-diol with 99% ee. PMID- 11871913 TI - Preparation of enantiopure ketones and alcohols containing a quaternary stereocenter through parallel kinetic resolution of beta-keto nitriles. AB - Racemic 1-methyl-2-oxocycloalkanecarbonitriles have been subjected to bioreduction by the fungus Mortierella isabellina NRRL 1757 through a parallel kinetic-resolution process. The u and l alcohols thus obtained (up to >99% ee) were easily separated and oxidized to the R and S ketones, respectively. The process can be then repeated so that both enantiomers of the ketone and two epimers of the alcohol can be obtained in their enantiopure forms. PMID- 11871914 TI - Electrogenerated base-induced N-acylation of chiral oxazolidin-2-ones. Part 2. AB - An improved and efficient electrochemical N-acylation of chiral oxazolidin-2-ones has been achieved. The generation of the nitrogen anion is obtained under mild conditions and without addition of base or probase, by direct electrolysis of a solution of MeCN-TEAP containing the oxazolidinone. Acylation agents (acid chlorides or anhydrides) were added at the end of the electrolysis. N-Acylated products were isolated in high to excellent yields. PMID- 11871915 TI - A simple and efficient method for the resolution of all four diastereomers of 4,4,4-trifluorovaline and 5,5,5-trifluoroleucine. PMID- 11871917 TI - The changing face of hypertension: is systolic blood pressure the final answer? PMID- 11871918 TI - Psychological factors in heart failure: a review of the literature. AB - Congestive heart failure (CHF) is the end stage of many diseases of the heart and a major cause of morbidity and mortality. The incidence of CHF is increasing steadily as treatment for its coronary antecedents, such as myocardial infarction, advances. Treatment of CHF generally relies on a battery of pharmacological interventions, alongside exercise and diet regimens. It is only in recent years that the psychological impact of heart failure has been explored, which is reflected by the absence of standardized psychological assessment for patients with CHF. In this article, we review studies that have addressed the effects of depression, anxiety, coping style, and level of social support in CHF. From the available evidence, it appears that patients generally experience moderate levels of depression, but not greatly heightened anxiety. Level of social support and style of coping with the disease are, however, important prognostic factors. It is difficult to draw definitive conclusions owing to the paucity of literature. Further work examining this issue is needed if the psychological issues of heart failure are not to be neglected. PMID- 11871919 TI - Clostridial myonecrosis cluster among injection drug users: a molecular epidemiology investigation. AB - A molecular epidemiologic investigation was performed on a cluster of severe necrotizing Clostridium infections in 5 injection drug users admitted to an urban community hospital. Interviews with survivors suggested a point source of infection. Pulsed-field gel electrophoresis of SmaI restriction digests was performed to determine the molecular relatedness of clinically obtained isolates and isolates obtained from heroin samples and the home environment. A common clonal strain was found in Clostridium sordellii isolates from 2 socially unrelated patients and from drug paraphernalia. Clonality of a Clostridium perfringens strain from another patient isolate was identical to an isolate from a syringe found in her home. Other C perfringens isolates from patients, heroin, and the environment were determined to be polyclonal. We postulate that rapid recognition and public health notification led to rapid resolution of the outbreak. PMID- 11871920 TI - Do subspecialists working outside of their specialty provide less efficient and lower-quality care to hospitalized patients than do primary care physicians? AB - BACKGROUND: Studies show that subspecialists can provide better quality care than primary care physicians when working within their subspecialty for patients with some medical conditions. However, many subspecialists care for patients outside of their chosen subspecialty. The present study compared the quality of care provided by subspecialists practicing outside of their specialty, general internists, and subspecialists practicing within their specialty. METHODS: The severity-adjusted mortality rate and the severity-adjusted length of stay were used as indexes of quality of care. Data from 5112 hospital admissions (301 different physicians) for community-acquired pneumonia, acute myocardial infarction, congestive heart failure, or upper gastrointestinal hemorrhage at 6 hospitals in the greater Cleveland, Ohio, area were used in this study. The data were severity adjusted with the CHOICE Severity of Illness System. RESULTS: Subspecialists working outside of their subspecialty cared for 25% of hospitalized patients. When comparing patients cared for by subspecialists practicing outside of their subspecialty, severity-adjusted lengths of stay were longer for patients with congestive heart failure (23% longer; 95% confidence interval [CI], 15%-32%), upper gastrointestinal hemorrhage (22% longer; 95% CI, 7%-39%), and community-acquired pneumonia (14% longer; 95% CI, 5%-24%) than for patients cared for by subspecialists practicing within their subspecialty. Patients also had a slightly higher hospital mortality rate when cared for by subspecialists practicing outside of their specialty than by subspecialists practicing within their subspecialty (mortality rate odds ratio, 1.46; P =.047). In addition, patients cared for by subspecialists practicing outside of their subspecialty had longer lengths of stay, and prolongations of stay were observed for patients with congestive heart failure (16% longer; 95% CI, 8%-26%), upper gastrointestinal hemorrhage (15% longer; 95% CI, 2%-30%), and community-acquired pneumonia (18% longer; 95% CI, 9%-28%) than patients cared for by general internists. CONCLUSIONS: Subspecialists commonly care for patients outside of their subspecialty, despite the fact that their patients may have longer lengths of stay than those cared for by subspecialists practicing within their specialty or by general internists. In addition, such patients may have slightly higher mortality rates than those cared for by subspecialists practicing within their subspecialty. PMID- 11871921 TI - Statin use, bone mineral density, and fracture risk: Geelong Osteoporosis Study. AB - BACKGROUND: Recent data suggest that 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors (statins) decrease fracture risk and increase bone mineral density (BMD). METHODS: This cross-sectional study is set in southeastern Australia. We evaluated the association between statin use, fracture risk, and BMD in 1375 women (573 with incident fractures and 802 without incident fracture, all drawn from the same community). Fractures were identified radiologically. Medication use and lifestyle factors were documented by questionnaire. RESULTS: Unadjusted odds ratio for fracture associated with statin use was 0.40 (95% confidence interval [CI], 0.23-0.71). Adjusting for BMD at the femoral neck, spine, and whole body increased the odds ratio to 0.45 (95% CI, 0.25-0.80), 0.42 (95% CI, 0.24-0.75), and 0.43 (95% CI, 0.24-0.78), respectively. Adjusting for age, weight, concurrent medications, and lifestyle factors had no substantial effect on the odds ratio for fracture. Statin use was associated with a 3% greater adjusted BMD at the femoral neck (P =.08), and BMD tended to be greater at the spine and whole body but did not achieve statistical significance. CONCLUSION: The substantial 60% reduction in fracture risk associated with statin use is greater than would be expected from increases in BMD alone. PMID- 11871922 TI - Balancing the risks of stroke and upper gastrointestinal tract bleeding in older patients with atrial fibrillation. AB - OBJECTIVE: To determine how factors that increase the risk of major upper gastrointestinal (GI) tract hemorrhage (recent upper GI tract bleeding or concurrent use of nonsteroidal anti-inflammatory drugs) influence the choice of antithrombotic therapy in older patients (those > or = 65 years) with atrial fibrillation. METHODS: For older patients with atrial fibrillation and no other contraindications to antithrombotic therapy, a Markov decision-analytic model was used to determine the preferred treatment strategy (no antithrombotic therapy, long-term aspirin use, or long-term warfarin sodium use) based on their risk of major upper GI tract hemorrhage. Input data were obtained by a systematic review of MEDLINE. Outcomes were expressed as quality-adjusted life-years (QALYs). RESULTS: For 65-year-old patients with average risks of stroke and upper GI tract bleeding, warfarin therapy was associated with 12.1 QALYs per patient; aspirin therapy, 10.8 QALYs; and no antithrombotic therapy, 10.1 QALYs. For persons with significantly higher risks of upper GI tract bleeding and/or lower risks of stroke, warfarin was no longer clearly the optimal antithrombotic therapy (eg, for 80-year-old persons with a baseline risk of stroke of 4.3% per year who were concurrently taking a conventional nonsteroidal anti-inflammatory drug: warfarin, 7.44 QALYs; aspirin, 7.39 QALYs; and no treatment, 7.21 QALYs). CONCLUSIONS: For older patients with atrial fibrillation and factors that place them at a higher than average risk of upper GI tract bleeding, the optimal choice of antithrombotic therapy to prevent stroke can vary according to the magnitude of this risk. Based on the risks of stroke and upper GI tract bleeding, clinicians can use the treatment recommendations of this study to provide rational stroke prevention therapy for older patients with atrial fibrillation. PMID- 11871923 TI - Quality of life after acute myocardial infarction among patients treated at sites with and without on-site availability of angiography. AB - BACKGROUND: Previous studies have compared the treatment and outcome of patients with acute myocardial infarction (AMI) admitted at sites with and without availability of angiography. Although mortality rates do not differ, it is unknown if quality of life (QOL) and functional status differ. METHODS: We measured QOL and functional status in patients with AMI treated within Quebec at 5 sites with (n = 253) and 5 sites without (n = 334) angiography. RESULTS: At admission, clinical characteristics, complication rates, and baseline measures of QOL and functional status were similar at sites with and without angiography. During hospitalization, patients treated at sites with angiography were more likely to undergo an invasive cardiac procedure than patients admitted at sites without angiography (angiography, 63% vs 26%; percutaneous transluminal coronary angioplasty, 33% vs 13%; and coronary artery bypass graft, 12% vs 5%). At 30 days and 6 months after AMI, QOL was slightly superior at sites with angiography, but by 1 year, most measures of QOL were back to baseline at both types of sites and were similar between the 2 groups. At 6 months, most standard health-related QOL components were similar; only physical and emotional role limitations were higher at sites with angiography. Return to work occurred earlier (at 30 days, 23% vs 12%), and a lower proportion of patients was readmitted for angina (within 1 year after AMI, 12% vs 18%) at sites with angiography. CONCLUSIONS: In the early post AMI period, the QOL of patients admitted at sites with angiography was higher than that of patients admitted at sites without angiography. However, by 1 year, the QOL and functional status of patients was similar in both groups. Differences in QOL were greatest when differences in treatment were greatest, lending support to a positive albeit small association between an early invasive approach to post AMI care and improved QOL. PMID- 11871924 TI - Effectiveness of thrombolytic therapy for acute myocardial infarction in the elderly: cause for concern in the old-old. AB - BACKGROUND: National guidelines have encouraged increased use of thrombolytic therapy for elderly patients with acute myocardial infarction (AMI). However, evidence supporting thrombolytic therapy in patients 75 years and older is lacking. In a retrospective cohort study of 2659 elderly AMI patients, we determined the association between thrombolytic use and in-hospital mortality by age and among patients with or without absolute or relative contraindications to thrombolytic treatment. METHODS: We abstracted the medical records of 2659 elderly patients admitted with AMI at 37 Minnesota community hospitals between 1992 and 1996. The main outcome measure was in-hospital mortality, controlling for demographic, clinical, comorbidity, and severity-of-illness variables. RESULTS: Sixty-three percent of 719 eligible patients received thrombolytic therapy. Twenty-seven percent of thrombolytic recipients had absolute contraindications to treatment. Patients receiving thrombolytic agents had fewer and less severe comorbidities than those not receiving thrombolytic therapy. There was a 4% increase in the odds of death for every 1-year increase in age for all thrombolytic recipients vs nonrecipients (odds ratio [OR], 1.04 per year; 95% confidence interval [CI], 1.01-1.08; P =.03). Among patients with 1 or more contraindication, the OR for death associated with thrombolytic use was 1.57 (95% CI, 1.03-2.40; P =.04). The adjusted odds of death among eligible thrombolytic recipients (vs nonrecipients) increased significantly with age (OR, 1.08 per year; 95% CI, 1.02-1.14; P =.008). Among eligible patients aged 80 to 90 years, the predicted odds of death among thrombolytic recipients vs nonrecipients was 1.4. Among eligible patients younger than 80 years, thrombolytic use was associated with reduced mortality. CONCLUSIONS: Our findings suggest the need for more research on the effectiveness of thrombolytic therapy for AMI patients 75 years and older and for more careful selection of elderly patients for this treatment. PMID- 11871925 TI - Prospective study of moderate alcohol consumption and risk of hypertension in young women. AB - BACKGROUND: Heavy alcohol consumption is associated with an increased risk of hypertension. However, the effect of moderate alcohol consumption; the specific effects of wine, beer, and liquor; and the pattern of drinking in relation to risk of hypertension among young women are unclear. METHODS: We prospectively examined the association between alcohol consumption and subsequent risk of hypertension among 70 891 women 25 to 42 years of age. RESULTS: During the 8 years of follow-up, 4188 cases (5.9%) of incident hypertension were reported. After adjustment for multiple covariates, the association between alcohol consumption and risk of hypertension followed a J-shaped curve. Compared with nondrinkers, the risk of developing hypertension according to average number of drinks consumed per day was as follows: 0.25 or less, 0.96 (95% confidence interval [CI], 0.89-1.03); 0.26 to 0.50, 0.86 (95% CI, 0.75-0.98); 0.51 to 1.00, 0.92 (95% CI, 0.82-1.04); 1.01 to 1.50, 1.00 (95% CI, 0.80-1.24); 1.51 to 2.00, 1.20 (95% CI, 0.92-1.58); and more than 2.0 drinks, 1.31 (95% CI, 1.02-1.68). Exclusion of past drinkers yielded similar results. Among women in the highest category of alcohol consumption, there was a suggestion that the increased risk of hypertension was present regardless of the specific beverage consumed (beer, wine, or liquor). Episodic drinking, defined as consumption of more than 10.5 drinks over 3 or fewer days per week, was not associated with increased risk of hypertension (relative risk, 0.80; 95% CI, 0.51-1.23). CONCLUSIONS: The association between alcohol consumption and risk of chronic hypertension in young women follows a J-shaped curve, with light drinkers demonstrating a modest decrease in risk and more regular heavy drinkers demonstrating an increase in risk. PMID- 11871926 TI - Prognostic value of systolic and diastolic blood pressure in treated hypertensive men. AB - BACKGROUND: The aim of this study was to assess the cardiovascular risk in hypertensive subjects according to systolic blood pressure (SBP) and diastolic blood pressure (DBP) levels. METHODS: The study sample consisted of 4714 hypertensive men, treated by their physician, who had a standard health checkup at the d'Investigations Preventives et Cliniques Center, Paris, France, between 1972 and 1988. Cardiovascular disease (CVD) and coronary heart disease (CHD) mortality were assessed for a mean period of 14 years. RESULTS: Among treated subjects, 85.5% presented uncontrolled values for SBP (> or = 40 mm Hg) and/or DBP (> or = 90 mm Hg). After adjustment for age and associated risk factors, these subjects presented an increased risk for CVD mortality (risk ratio [RR], 1.66; 95% confidence interval [CI], 1.04-2.64) and for CHD mortality (RR, 2.35; 95% CI, 1.03-5.35) compared with controlled subjects. After adjustment for age, associated risk factors, and DBP, and compared with subjects with SBP under 140 mm Hg, the RR for CVD mortality was 1.81 (95% CI, 1.04-3.13) in subjects with SBP between 140 and 160 mm Hg and 1.94 (95% CI, 1.10-3.43) in subjects with SBP over 160 mm Hg. By contrast, after adjustment for SBP levels, CVD risk was not associated with DBP. Compared with subjects with DBP under 90 mm Hg, RR for CVD mortality was 1.17 (95% CI, 0.80-1.70) in subjects with DBP between 90 and 99 mm Hg and 1.03 (95% CI, 0.67-1.56) in subjects with DBP over 100 mm Hg. Similar results were observed for CHD mortality. CONCLUSIONS: In hypertensive men treated in clinical practice, SBP is a good predictor of CVD and CHD risk. Diastolic blood pressure, which remains the main criterion used by most physicians to determine drug efficacy, appears to be of little value in determining cardiovascular risk. Evaluation of risk in treated individuals should take SBP rather than DBP values into account. PMID- 11871927 TI - Systolic vs diastolic blood pressure control in the hypertensive patients of the PAMELA population. Pressioni Arteriose Monitorate E Loro Associazioni. AB - BACKGROUND: Previous studies have shown that in the treated fraction of the hypertensive population, blood pressure (BP) control is less common for systolic BP (SBP) than for diastolic BP (DBP) as measured in the physician's office. Whether this phenomenon is artifactually attributable to a temporary increase in BP owing to a "white-coat" effect or represents a true rarity of SBP control in daily life is unknown. METHODS: Data were obtained from the PAMELA (Pressioni Arteriose Monitorate E Loro Associazioni) study population, which involved individuals ranging in age from 25 to 74 years who were representative of the residents of Monza (a city near Milan, Italy) and who were stratified according to sex. Office (an average of 3 sphygmomanometric measurements), home (an average of morning and evening self-measurements using a semiautomatic device), and 24 hour ambulatory (average of measurements performed every 20 minutes during the day and at night) BP values were obtained in all study subjects. In the treated hypertensive patients, BP was regarded as controlled if office values were less than 140 (SBP) or 90 (DBP) mm Hg. Home and 24-hour average SBP and DBP were regarded as controlled if the values were lower than 132/83 and 125/79 mm Hg, respectively. RESULTS: In the study participants (n = 2051), the number of patients with hypertension who were receiving antihypertensive treatment was 398, or approximately 42% of all individuals with hypertension. In-office SBP control by treatment was less frequent than DBP control (29.9% vs 41.5%, P<.05). This was also the case when home and 24-hour SBP and DBP control was considered (38.3% vs 54.6% and 50.8 vs 64.9%, respectively, P<.05 for both). CONCLUSIONS: In the PAMELA population, SBP control by treatment was much less frequent than DBP control by treatment. This was the case not only for office BP values but also for home and 24-hour BP values, demonstrating that inadequate SBP control is not limited to artificial BP-measuring methods but occurs in daily life. PMID- 11871928 TI - The association of sex and payer status on management and subsequent survival in acute myocardial infarction. AB - BACKGROUND: Previous reports have generally shown lower utilization of hospital resources and lower survival in women than men with acute myocardial infarction. However, to our knowledge, no reports have described the influence of payer status on the treatment and outcome of women and men with acute myocardial infarction. METHODS: Baseline and clinical presenting characteristics, utilization of hospital resources, and subsequent clinical outcome were ascertained among 327 040 women and men enrolled in a national registry of myocardial infarction from June 1, 1994, to January 31, 1997. Separate Cox regression analyses were performed for Medicare, Medicaid, health maintenance organizations, and commercial payer groups to ascertain variables that were predictive of mortality in the study population. RESULTS: After adjustment for differences in age and other baseline and presenting characteristics, women were significantly more likely than men to die in the hospital (hazard ratio, 1.13; 95% confidence interval, 1.10-1.16), and this difference was greatest among women with health maintenance organization and commercial insurance (hazard ratios, 1.30 and 1.29, respectively), and least among women with Medicare (hazard ratio, 1.07). However, after adjustment for the additional effect on short-term survival of sex differences in the utilization of both pharmacologic treatments administered within the first 24 hours and invasive cardiac procedures, the mortality difference observed for women and men further diminished (hazard ratio, 1.08; 95% confidence interval, 1.05-1.10). CONCLUSION: In this large registry, we did not observe significant variations among payer classes in management and mortality among women and men after acute myocardial infarction. PMID- 11871929 TI - Association between Chlamydia pneumoniae antibodies and intimal calcification in femoral arteries of nondiabetic patients. AB - BACKGROUND: Chlamydia pneumoniae, a gram-negative bacterium, has been suggested to be a risk factor for atherosclerosis. Calcium is a well-known component of atherosclerotic plaques, but it is uncertain whether infectious agents play a role in the calcification process of the arteries. PATIENTS: To address this issue we investigated the association of Chlamydia antibodies with intimal arterial calcification as assessed by soft tissue radiograms from the thigh region of 1373 nondiabetic Finnish individuals aged 45 to 64 years. RESULTS: At baseline, radiologically detectable intimal calcification in femoral arteries was found in 172 (27%) of 638 men and 43 (6%) of 735 women (P<.001). The presence of intimal artery calcifications was strongly related to conventional atherosclerotic risk factors and to Chlamydia antibodies. In Cox regression analysis, association of Chlamydia antibodies with intimal artery calcification persisted after extensive adjustment for other cardiovascular risk factors (P =.04). A dose-response relationship was observed between Chlamydia antibodies and intimal femoral artery calcification (P =.006). The presence of intimal artery calcification was strongly associated with an increased risk of future coronary heart disease mortality (P<.001). CONCLUSION: Chlamydia antibodies are strongly associated with intimal calcification of the femoral arteries. PMID- 11871930 TI - The impact of empirical management of acute cystitis on unnecessary antibiotic use. AB - BACKGROUND: Guidelines for the management of acute cystitis support empirical antibiotic treatment; however, up to half of symptomatic women have negative urine cultures. OBJECTIVE: To determine whether empirical treatment leads to unnecessary antibiotic prescriptions in women with symptoms of acute cystitis. METHODS: A cohort of 231 women (defined as females aged 16 years and older) presenting to family physicians' offices with symptoms of cystitis underwent a standardized clinical assessment, urine dip testing, and culture. Recommendations for urine testing and antibiotic treatment under 3 empirical strategies were compared with observed physician management and a logistic regression model for the outcomes of antibiotic prescriptions, urine culture testing, and unnecessary antibiotics, defined as a prescription where the subsequent urine culture was negative. RESULTS: There were 123 positive urine cultures (53.3%). Physicians prescribed antibiotics to 186 women (80.9%), of whom 74 (39.8%) were culture negative. Unnecessary antibiotic use was similar for 2 guidelines recommending empirical antibiotic treatment without testing for pyuria (41.4% and 40.6%). Treating women with classic cystitis symptoms and pyuria would have decreased unnecessary antibiotic use (26.2%; P =.02) but resulted in fewer women with confirmed urinary tract infection receiving immediate antibiotics (66.4% vs 91.8% usual care; P<.001). A derived prediction model incorporating testing for pyuria and nitrites would also have reduced unnecessary antibiotic use (27.5%; P =.03), but more women with confirmed urinary tract infection would have received immediate antibiotics (81.3%; P =.01). CONCLUSIONS: Empirical antibiotic treatment of acute cystitis in women without testing for pyuria promotes unnecessary antibiotic use. A simple decision rule provides for prompt treatment of infected women while reducing antibiotic overuse and unnecessary urine testing. PMID- 11871931 TI - Bruxism masquerading as a murmur. PMID- 11871932 TI - West Nile virus meningoencephalitis with optic neuritis. PMID- 11871933 TI - Nationwide folate fortification has complex ramifications and requires careful monitoring over time. PMID- 11871934 TI - Underestimation of adverse drug events in nursing home residents. PMID- 11871936 TI - Blood pressure in early life and cardiovascular disease mortality. PMID- 11871938 TI - Predicting tuberculosis at hospital admission. PMID- 11871941 TI - Testing for hypercoagulable disorders. PMID- 11871940 TI - The effect of explicit financial incentives on physician behavior. PMID- 11871942 TI - Timing of hypercoagulable screens. PMID- 11871945 TI - Predicting difficult intubation--worthwhile exercise or pointless ritual? PMID- 11871946 TI - Effects of desflurane and isoflurane on intestinal tissue oxygen pressure during colorectal surgery. AB - Volatile anaesthetics differ in the effects they have on splanchnic haemodynamics and oxygenation. The aim of this study was to evaluate the effects of desflurane and isoflurane as part of a balanced anaesthetic technique on intestinal tissue oxygenation during colorectal surgery. Data were analysed from 44 patients randomly assigned to receive either desflurane (desflurane group, n = 20), or isoflurane (isoflurane group, n = 24) for inhalational anaesthesia. Tissue oxygen pressure (P(tiss)O2) was measured on the serosal side of the large intestine prior to colonic resection (T1) and following the completion of the bowel anastomosis (T2). In addition, haemodynamic and oxygenation parameters were assessed. No difference in mean P(tiss)O2 was observed between the groups at T1 [desflurane group: 8.1 (2.9) kPa vs. isoflurane group: 7.7 (2.7) kPa]. Following completion of the anastomosis (T2) mean P(tiss)O2 was higher in the isoflurane group [9.6 (2.9) kPa] than the desflurane group [7.7 (2.4) kPa, p = 0.025]. During surgery no difference between the groups could be observed with regard to haemodynamics and global oxygenation parameters. The lack of a difference between the groups in P(tiss)O2 before resection of the colon suggests that, under normal conditions, desflurane and isoflurane have comparable effects on intestinal blood flow and oxygenation. However, following local ischaemia, the reactive hyperaemia seems to be better preserved during isoflurane anaesthesia indicated by a local increase in P(tiss)O2blank(p = 0.013) following completion of the bowel anastomosis. PMID- 11871947 TI - Personality testing and profiling for anaesthetic job recruitment: attitudes of anaesthetic specialists/consultants in New Zealand and Scotland. AB - Specialist/consultant anaesthetists based in New Zealand and Scotland were sent a reply paid postal questionnaire asking about their attitudes to personality testing and personality types in the recruitment process for registrars and specialists. The questionnaire consisted of nine Likert-style questions and 14 visual analogue questions. The overall response rate was 65% (523/808). The responses to all the questions were broadly similar in the two countries. Personality testing was deemed of use in recruiting trainees and specialists, with a slightly greater proportion considering personality traits more important than academic achievement. An overwhelming majority believed the presence of an adverse personality trait would influence an appointment process, but few believed that the personality makeup of anaesthetists influenced the way in which they react to stressful situations. A slight majority considered the interview process a poor predictor of personality. New Zealand anaesthetists rated independence, orderliness, compassion, empathy, reflectiveness and patience higher than did anaesthetists in Scotland. In contrast, anaesthetists in Scotland rated pragmatism, as opposed to perfection, as a more important characteristic than did the New Zealand specialists. Personality assessment, although not effective as the sole tool for candidate selection, may have a role in the process of anaesthetic job recruitment and warrants further investigation. PMID- 11871948 TI - Orotracheal fibreoptic intubation for rapid sequence induction of anaesthesia. AB - We assessed whether flexible fibreoptic-guided orotracheal intubation could be rapidly and successfully achieved during a simulated rapid sequence induction in 30 anaesthetised and paralysed patients. Rapid sequence induction was simulated by applying practised cricoid pressure. Using a flexible fibreoptic laryngoscope with camera and closed circuit television, an anaesthetist experienced with the technique performed orotracheal endoscopy and intubation with a cuffed 7.0-mm Portex tracheal tube through a VBM Bronchoscope Airway. Fibreoptic intubation was successful at the first attempt in 28 patients (93%); two patients required two attempts. Mean (SD) time from removal of the facemask from the patient's face to the appearance of carbon dioxide in the expired breath after intubation was 111 (46) s (median 100 s; range 54-195 s). There were one or more difficulties in 13 patients (43%). These difficulties were largely avoidable and included problems with fibreoptic equipment, the Bronchoscope Airway, copious secretions, cricoid pressure or railroading of the tracheal tube. Flexible fibreoptic-guided orotracheal intubation may have a place in the management of failed intubation during a rapid sequence induction. PMID- 11871949 TI - Comparison of times to achieve tracheal intubation with three techniques using the laryngeal or intubating laryngeal mask airway. AB - We compared the times to intubate the trachea using three techniques in 60 healthy patients with normal airways: (i) fibreoptic intubation with a 6.0-mm reinforced tracheal tube through a standard laryngeal mask airway (laryngeal mask fibreoptic group); (ii) fibreoptic intubation with a dedicated 7.0-mm silicone tracheal tube through the intubating laryngeal mask airway (intubating laryngeal mask-fibreoptic group); (iii) blind intubation with the dedicated 7.0-mm silicone tracheal tube through the intubating laryngeal mask airway (intubating laryngeal mask-blind group). Mean (SD) total intubation times were significantly shorter in the intubating laryngeal mask-blind group (49 (20) s) than in either of the other two groups (intubating laryngeal mask-fibreoptic 74 (21) s; laryngeal mask fibreoptic group 75 (36) s; p < 0.001). However, intubation at the first attempt was less successful with the intubating laryngeal mask-blind technique (15/20 (75%)) than in the other two groups (intubating laryngeal mask-fibreoptic 19/20 (95%) and laryngeal mask-fibreoptic 16/20 (80%)) although these differences were not statistically significant. We conclude that in this patient group, all three techniques yield acceptable results. If there is a choice of techniques available, the intubating laryngeal mask-blind technique would result in the shortest intubation time. PMID- 11871951 TI - Anions and the anaesthetist. AB - Anions are the negative components of most chemical structures and play many important physiological and pharmacological roles that are of interest to the anaesthetist. Their relevance is reviewed with a particular emphasis on the inorganic anions (halides, bicarbonate, phosphate and sulphate) and the significance and limitations of the anion gap. Organic anions (albumin, lactate) are also discussed, albeit briefly. The suitability of anions for their role in neurotransmission and acid-base balance is outlined. PMID- 11871950 TI - Remifentanil and the tunnelling phase of paediatric ventriculoperitoneal shunt insertion. A double-blind, randomised, prospective study. AB - Sixty-two children were randomly allocated to receive, during inhalational anaesthesia with isoflurane and nitrous oxide, either 1.0 microg x kg(-1) remifentanil (n = 33) or saline (n = 29) just before the tunnelling phase of ventriculoperitoneal shunt insertion, in a double-blind study. The remifentanil group showed little stress response to tunnelling as indicated by median (interquartile range [range]) change in heart rate -5.2 (-11.4 to 9.8 [-19.4 to 30.4])%, mean arterial pressure -5.0 (-20.8 to 15.5 [-40.9 to 42.9])% or plasma norepinephrine -13.5 (-38.1 to -2.5 [-77.7 to 81.5])% compared with the saline group, in which the changes were 20.1 (11.5-36.1 [2.1-83.1])%, 42.7 (27.1-56.8 [3.2-73.5])% and 13.3 (0.8-70.0 [-45.2 to 337.5])%, respectively (p < 0.001 for all comparisons). These changes were consistent across most different age categories. The cardiovascular response in the saline group lasted for 8 (4-15 [0 39]) min. Tracheal extubation occurred after 3 (2-4 [1-8]) min in the remifentanil group and 3 (2-6 [0-15]) min in the saline group (p = 0.29), with transfer to the recovery area and discharge to the ward, respectively, 4 (4-5 [1 10]) min and 9 (7-13 [2-32]) min in the remifentanil group and 7 (4-8 [2-18]) min and 14 (10-19 [7-44]) min in the saline group (p = 0.06 and 0.01, respectively). Postoperatively there was some evidence of respiratory depression and increased oxygen requirements in all age categories, but this was similar in both groups. Overall, the maximum increase from baseline in transcutaneous carbon dioxide tension was 41.2 (11.3-66.7 [-2.0 to 141.7])% in the remifentanil group compared with 30.7 (20.5-55.1 [1.7-159])% in the saline group (p = 0.8), and the time taken for transcutaneous carbon dioxide tension to decrease to < 6.0 kPa was 4 (0 13 [0-60]) min compared with 7 (0-13 [0-60]) min, respectively (p = 0.75). There was no difference between the two groups in postoperative analgesic requirements or in blood loss and there were no significant side-effects. We conclude that remifentanil is an appropriate and safe analgesic to provide balanced anaesthesia to cover the tunnelling phase of paediatric ventriculoperitoneal shunt insertion. PMID- 11871952 TI - Tracheal intubation and sore throat: a mechanical explanation. AB - Although tracheal intubation remains a valuable tool, it may result in pressure trauma and sore throat. The evidence for an association between these sequelae is not conclusive and sore throat may be caused at the time of intubation. This hypothesis was tested in a mechanical model and the results from tracheal intubation compared with those from insertion of a laryngeal mask airway, which is associated with a lower incidence of sore throat. Use of the model suggests that the tracheal tube and laryngeal mask airway impinge on the pharyngeal wall in different manners and involve different mechanisms for their conformation to the upper airway, but that in a static situation, the forces exerted on the pharyngeal wall are low with both devices. It also suggests that the incidence of sore throat should be lower for softer and smaller tracheal tubes and that the standard 'Magill' curve (radius of curvature 140 +/- 20 mm) is about optimum for the average airway. PMID- 11871953 TI - Measuring the filtration performance of breathing system filters using sodium chloride particles. AB - The filtration performance of 33 breathing system filters (nine pleated hydrophobic and 24 electrostatic filters) was measured using sodium chloride particles. The particles had a size distribution with a count median diameter of 0.07 microm and a geometric standard deviation not exceeding 1.83. The geometric mean penetration values ranged from 0.002 to 0.67% for the nine pleated hydrophobic filters and from 0.25 to 35% for the 24 electrostatic filters (p < 0.0001 for the difference between the two filter types). The filtration performance obtained when filters are challenged with either sodium chloride particles or microbes is compared and discussed. PMID- 11871954 TI - The effect of 10 degrees head-up tilt in the right lateral position on the systemic blood pressure after subarachnoid block for Caesarean section. AB - Forty women presenting for elective Caesarean section under spinal anaesthesia were randomly assigned to have anaesthesia induced in the right lateral position either in the horizontal position or with 10 degrees head-up tilt. Hyperbaric bupivacaine 2 ml 0.5% with 0.1 mg of morphine was injected intrathecally before the parturients were placed in the supine position with 15 degrees left lateral tilt. Blood pressure and heart rate were monitored every minute and the sensory level (loss of sharp sensation to pinprick) was monitored every 3 min until clamping of the umbilical cord. Ephedrine 6 mg was given every minute that the systolic blood pressure decreased below 90 mmHg. The mean systolic blood pressure during the first 5 min after induction of spinal anaesthesia was lower in the control group compared to the tilted group (99 mmHg vs. 109 mmHg; p = 0.043). The upper limit of block was higher in the control group compared to the tilted group (p = 0.002). The use of 10 degrees head-up tilt resulted in a reduced incidence of hypotension initially and less extensive sensory block. PMID- 11871955 TI - The margin of safety associated with the use of cuffed paediatric tracheal tubes. AB - There is increasing interest in the use of cuffed paediatric tracheal tubes, these tubes have a number of advantages and disadvantages. One disadvantage is the decreased margin of safety associated with the use of cuffed tracheal tubes. When using an uncuffed tube, the margin of safety is approximately the length of the trachea, however, for a cuffed tube, this margin is reduced and is the length of the trachea minus the distance between the proximal edge of the cuff and the and the tip of the tube. We have found that, compared with traditional uncuffed tubes, cuffed tubes may be associated with a reduced margin of safety of approximately 50% or more. PMID- 11871956 TI - Day surgery and body mass index: results of a national survey. AB - In March 1992, the Royal College of Surgeons issued Guidelines for Day Case Surgery. Patients with a body mass index (BMI) > 30 were deemed unsuitable for operations to be performed as a day case. Since these guidelines were issued, many changes have occurred. Two years ago we successfully increased the BMI limit for patients undergoing general anaesthesia in our day surgery unit from 30 to 34. The success of this led us to question the current validity of the Royal College of Surgeons guidelines. A postal questionnaire was conducted surveying current practice in day surgical units within the UK. We achieved a 96% response rate. The results demonstrated a range of acceptable BMI values, with 85% of units anaesthetising patients with a BMI > 30. We conclude that many day case units routinely anaesthetise patients with BMI values > 30. Therefore, the current guidelines, which were issued 9 years ago, are no longer being adhered to nationally. PMID- 11871957 TI - Effect of videotape feedback on anaesthetists' performance while managing simulated anaesthetic crises: a multicentre study. AB - The aim of this study was to examine the performance of anaesthetists while managing simulated anaesthetic crises and to see whether their performance was improved by reviewing their own performances recorded on videotape. Thirty-two subjects from four hospitals were allocated randomly to one of two groups, with each subject completing five simulations in a single session. Individuals in the first group completed five simulations with only a short discussion between each simulation. Those in the second group were allowed to review their own performance on videotape between each of the simulations. Performance was measured by both 'time to solve the problem' and mental workload, using anaesthetic chart error as a secondary task. Those trainees exposed to videotape feedback had a shorter median 'time to solve' and a smaller decrease in chart error when compared to those not exposed to video feedback. However, the differences were not statistically significant, confirming the difficulties encountered by other groups in designing valid tests of the performance of anaesthetists. PMID- 11871958 TI - Outreach critical care is not the solution. PMID- 11871959 TI - An unexpected complication resulting from the use of a laryngeal mask during an operation to remove a branchial cyst. PMID- 11871960 TI - Outreach critical care. PMID- 11871961 TI - Intensive care costs. PMID- 11871962 TI - Overcoming difficulties with percutaneous tracheostomy. PMID- 11871964 TI - Cricoid pressure application by intensive care nurses. PMID- 11871965 TI - Cardiac output determination using compliance. PMID- 11871966 TI - Manpower requirements when implementing a partial shift system for anaesthetic juniors. PMID- 11871967 TI - Probability of winning the National Lottery. PMID- 11871969 TI - Anaesthesia induction rooms--sheer luxury. PMID- 11871970 TI - Compartment syndrome. PMID- 11871971 TI - Obstruction of airway equipment. PMID- 11871972 TI - Another airway foreign body. PMID- 11871973 TI - Yet another foreign body in a laryngeal mask airway. PMID- 11871974 TI - A phantom capnograph trace. PMID- 11871975 TI - Phantom anaesthetic vapour. PMID- 11871977 TI - Anaesthetic machine safety--the story continues. PMID- 11871978 TI - Halothane vs. sevoflurane in the difficult airway. PMID- 11871980 TI - Novel use of capnography during an awake fibreoptic intubation. PMID- 11871981 TI - Arterial line insertion. PMID- 11871982 TI - Flush volume for a vascular access device. PMID- 11871983 TI - Experience of complications of percutaneous dilatational tracheostomy. PMID- 11871984 TI - Single-lung ventilation via a double lumen tube in a patient with a tracheostomy. PMID- 11871985 TI - Acid-base disorders in the critically ill. PMID- 11871987 TI - Inadvertent catheterisation of a partial anomalous pulmonary venous channel during central venous cannulation. PMID- 11871988 TI - Correct nomenclature for stereo isomers. PMID- 11871989 TI - Hypo-osmolar Hartmann's. PMID- 11871990 TI - Effective labelling is difficult, but safety really does matter. PMID- 11871993 TI - Cheese, drug labels and anaesthetic room error. PMID- 11871992 TI - Use of herbal medicines in ambulatory surgical patients. PMID- 11871994 TI - A-Q alphabetic anaesthetic assessment algorithm. PMID- 11871991 TI - Persistent cough following target-controlled infusion (TCI) with propofol. PMID- 11871995 TI - An unusual complication of fasting. PMID- 11871996 TI - Can propofol cause keratitis? PMID- 11871997 TI - The incidence of pulmonary oedema in fit patients of African origin. PMID- 11871998 TI - Preventing rubber stopper coring. PMID- 11871999 TI - Non-English information on ampoules and drug package-inserts. PMID- 11872001 TI - Just who do they think we are? PMID- 11872000 TI - Anaesthetic history of a patient: 250 anaesthetics in 30 years. PMID- 11872004 TI - Prions and blood: the impact on artificial organ technology development. PMID- 11872005 TI - The effect of methanol washing of plasticized polyvinyl chloride on biomaterial contact-mediated CD11b (mac-1) expression in a rat recirculation model. AB - Our objective was to assess whether using a methanol wash to reduce the level of plasticizer present on the surface of medical-grade polyvinyl chloride (PVC) has a moderating effect on the expression of CD11b (mac-1) on neutrophils in rats undergoing recirculation. The study was carried out on 3 groups of 10 adult male Sprague-Dawley rats weighing between 350 and 450 g. In the 2 test groups, the animals were exposed to 48 cm(2) of di-(2-ethyl-hexyl)-phthalate (DEHP) plasticized PVC in a parallel plate recirculating test cell through which blood was recirculated at 1.5 ml/min. In the first test group, the PVC was untreated; in the second test group, the PVC was washed in methanol to reduce the level of plasticizer on the surface. The test cell was connected to the right femoral circulation, and recirculation was established for a period of 60 min. Blood samples were taken at 0, 30, and 60 min for assessment of CD11b expression on neutrophils using flow cytometric analysis. In a third group of 10 control experiments, rats underwent the entire surgical procedure, but without recirculation through the test cell. There was statistically significant (p < 0.001) lower Cd11b expression on neutrophils in the blood of rats perfused through the cell containing methanol-washed PVC after 30 min and at 60 min. CD11b expression was significantly (p < 0.001) lower in the control group than in both test groups at both the 30 and 60 min time points and at the 60 min time point on comparison with the group where blood was perfused through methanol-washed PVC. These results demonstrate that the biomaterial-contact-mediated upregulation of CD11b may be significantly reduced by employing a methanol-washing technique on the plasticized PVC. Although this technique does not entirely eliminate the expression of CD11b on neutrophils, the difference is significant and suggests the role of the plasticizer in the development of this inappropriate inflammatory response. PMID- 11872006 TI - Atomic force microscopic observation of mechanically traumatized erythrocytes. AB - Erythrocytes are damaged or stimulated mechanically by artificial organs assisting in circulation. For several decades, a large number of research studies have been conducted to investigate the traumatizing phenomena due to nonphysiological flow conditions. These phenomena are thought to be the physical interaction between the cell membrane and the various fluidic conditions. To elucidate or evaluate the phenomena, however, chemical components emerging into the circulating solution, such as liberated hemoglobin or lactic dehydrogenase (LDH), have been measured as a main parameter. Naturally, the physical reaction caused on the membrane itself cannot be detailed by these parameters because they are the secondary products resulting from the mechanical membrane rupture. The aim of this study is to understand the traumatizing mechanism directly from a microbiological viewpoint. As a first step, we visualized the surface of sheep erythrocytes loaded with shear stress and measured erythrocyte surface roughness by atomic force microscopy (AFM) on a nanometer scale (10(-9) m). The constant shear rate was set at 1,800 (1/s), and the exposure time was set at 0.5, 1, and 2 h. We also measured the liberated hemoglobin concentration. As a result, it was found that the fine structure on the cell surface was changed drastically by the stress. It was also found that the surface roughness value increased with the exposure time, and correlated to the hemoglobin concentration. The visualization and the measurement of surface roughness of traumatized erythrocytes by AFM were thought to offer a new parameter for both hemolytic and subhemolytic studies. PMID- 11872007 TI - Effect of different solvents and crosslinkers on cytocompatibility of Type II collagen scaffolds for chondrocyte seeding. AB - Type II collagen extracted from porcine costal cartilage was evaluated as scaffolds for cartilage tissue engineering. Chemical crosslinkers were employed to improve the mechanical properties and the resistance toward degradation. Films and porous scaffolds were prepared from collagen solutions dissolved in 3% acetic acid (designated A) or in deionized water (designated W) and crosslinked by an epoxy (designated E) or by a carbodiimide (designated C). Immortalized rat chondrocytes and rabbit chondrocytes were used to assess cytocompatibility of crosslinked collagen matrices. Cell adhesion rate onto the films made by different preparations ranked in the order of WE > or = WC > AC > or = AE. Cell proliferation ranked in the order of AC > WC > AE > or = WE. Cells maintained round morphology only on AC and WC films. In 3-D seeding, AC scaffolds also were found to be the most cytocompatible. WC scaffolds, however, had better dimensional stability. It was concluded that Type II collagen scaffolds, when prepared by using deionized water as the solvent and carbodiimide as the crosslinker, could promote chondrocyte growth and matrix production. PMID- 11872008 TI - An investigation of blood damage induced by static pressure during shear-rate conditions. AB - Many investigators have studied the effect of a mechanical force (shear rate, pressure, or temperature) on hemolysis. However, there exists no investigation of a relationship between the interactions of mechanical forces and hemolysis. The purpose of this study is to investigate the interactions of mechanical forces on hemolysis. We performed in vitro tests by using bovine blood, applying shear rate (0, 500, 1,000, and 1,500 s-1), positive pressure (0, 200, 400, and 600 mm Hg), and temperature (21, 28, and 35 degrees C) simultaneously. In all temperatures at the shear rate of 1,500 s-1, there are statistically significant differences in the hemolysis rate between 0 and 600 mm Hg (p < 0.05). However, to investigate the effect of temperature on hemolysis, shear stress was calculated at each blood temperature. There were no statistically significant differences among them. The results suggested that erythrocyte trauma caused by pressure related to the level of shear rate. It was found that the causes of hemolysis included the shear rate as well as shear rate and pressure. PMID- 11872009 TI - In vitro investigation of opening behavior and hydrodynamics of bileaflet valves in the mitral position. AB - The performance of four 25 mm bileaflet valves of different designs was evaluated in the mitral position of our own pulse simulator. With the aid of a high-speed video camera, it was demonstrated that both the St. Jude Medical (SJM) valve (Hemodynamic Plus [HP] Series, St. Jude Medical, Inc., St. Paul, MN, U.S.A.) and the CM valve (CarboMedics, Inc., Austin, TX, U.S.A.) were able to open fully and that the CM valve fluttered much more vigorously at the fully open position than did the SJM HP valve. Conversely, neither the ATS valve (ATS Medical, Inc., Minneapolis, MN, U.S.A.) nor the On-X valve (Medical Carbon Research Institute, Austin, TX, U.S.A.) exhibited movement to a fully open configuration. The overall average opening angles of the ATS and the On-X, on 3, 4, and 5 L/min flow rate for a heart rate of 70 bpm and 5, 6, and 7 L/min for 100 bpm, were 74.8 degrees and 81.6 degrees, respectively, whereas their design opening angles were 85 degrees and 90 degrees. Pressure drops across the CM and the ATS were consistently higher than those of the On-X and the SJM HP. Closing volumes for all the valves were below 8% for a heart rate of 70 bpm. This in vitro investigation yielded the following conclusions: The ATS and On-X valves are not able to open fully in the mitral position, but this does not impair their normal function; both a larger orifice diameter and a large opening angle can decrease the pressure drop; in general, the On-X valve achieves its design goals in this experiment (i.e., it produces a lower pressure drop and lower closing volume by virtue of its large orifice and high-profile design); however, the hinge flow in the non-fully open state should be investigated further. PMID- 11872010 TI - Ripple reduction control of the undulation pump total artificial heart. AB - An undulation pump total artificial heart (UPTAH) in which the revolutions of the motor are converted to undulation motion of a disk has been developed. In an experiment, a goat using the UPTAH survived for 54 days. However, a large ripple was observed in the device's output pressure and flow waveform. In calculating the spectrum of the ripple, we found that the ripple mainly comprised 2 frequency sine waves: 1 having the same frequency as and 1 having double the frequency of the motor revolutions. To reduce the ripple, 2 sine waves, 1 having the same frequency as and 1 having double the frequency of the motor revolutions, were provided to the motor current to modulate the pulse width of the pulse width modulation controlling the motor revolutions. This ripple control method reduced the pressure ripple by 90% in a mock circulation and by 70% in animal experiments. These results revealed that the ripple generated in the UPTAH could be controlled through the use of motor control software. PMID- 11872011 TI - Inlet port positioning for a miniaturized centrifugal blood pump. AB - We are developing the Baylor-Kyocera KP implantable centrifugal blood pump for small sized adult and pediatric patients. This pump eccentrically positions the inlet port, which eliminates flow stagnation around the top pivot bearing. The inlet port design is important because it may vary the inlet orifice pressure on the top housing and change hydraulic performance and hemolytic characteristics. The pressure distribution inside the KP pump was assessed by a computational fluid dynamic (CFD) analysis with 2.7 x 10(5) elements and 3.16 x 10(5) nodes. Hydraulic performance and hemolysis were evaluated with 3 different pump housings, which had 3.8, 4.5, and 6.1 mm offset inlet ports from the center in a mock circuit. The CFD analysis revealed that the pressure gradually increased from the center toward the peripheral. The pressure difference between the 3.8 to 6.1 mm offsets was less than 600 Pa. The hydraulic performance did not drastically change at 3.8, 4.5, and 6.1 mm offset from the center. However, the hemolysis increased with the increase of the port offset from 0.0080+/- 0.0048 to 0.054 +/- 0.028 g/100 L. The inlet port positioning is important to attain less blood trauma in this small Gyro centrifugal blood pump. The preferable position of the inlet port is less than 4.5 mm offset from the center. PMID- 11872012 TI - Savings in dialysis treatment? AB - The steadily increasing number of dialysis patients prompts considerations on possibilities for budget reductions with maintenance of treatment quality. A literature survey is presented concerning trends of population increase, individual treatment costs, rationing of patient intake, and consequences of delayed progress of renal insufficiency as well as of savings during both the initial and the later phases of regular dialysis therapy. Cost reduction in one area may well induce rising total budgets and influence clinical outcome. A multidisciplinary approach is suggested to obtain answers to several questions: Can the economic burden of the changing patient demography be counterbalanced by a reorganized staff structure? Will early referral, good predialysis control, and incremental dialysis start imply longer survival? Will increased dialysis doses be economically neutralized by less staff requirements, drug consumption, and patient morbidity? Should dialyzer reuse be abandoned? Can pretransplant dialysis periods be reduced or omitted by improved planning? PMID- 11872013 TI - Serum neopterin monitoring and vitamin E-modified, regenerated hemodialyzer membrane influence on biocompatibility. AB - The exposure of blood to hemodialysis membranes results in numerous phenomena and/or complications in hemodialyzed patients, which have an influence on the quality of life (QOL) of those patients. A vitamin E-modified regenerated cellulose membrane (E-membrane) was developed to act as a scavenger for reactive oxygen species causing complications in hemodialysis patients. Neopterin (NEOP) is a metabolite derived from guanosine triphosphate with the production and release of NEOP being induced in monocytes and macrophages by cytokines such as interferon-gamma (IFN-gamma). Serum neopterin levels are shown to be a reactive marker of bioincompatibility of dialysis membranes in hemodialysis patients. The following report evaluates the usefulness of serum NEOP as a marker for the biocompatibility of the E-membrane hemodialyzer in a clinical study. In the clinical study, where extracorporeal ultrafiltration strategies with E-membranes were employed, the serum levels of NEOP were lower than those in patients using cellulose triacetate membranes (C-membranes). In the long-term evaluation of the biocompatibility of E- and C-membranes, the increase of serum neopterin levels in the C-membrane was higher than those in the E-membrane. In conclusion, the evaluation of serum neopterin levels during hemodialysis shows that the E membrane has a good biocompatibility in hemodialyzed patients. PMID- 11872014 TI - Light and electron microscopic analyses of autologous pericardial tissue used as a small-diameter arterial graft in dogs. AB - As a form of small-diameter arterial graft, we implanted fresh autologous pericardium and pericardium treated with 0.6% glutaraldehyde in the bilateral carotid arteries of dogs and then compared the time-related changes of the grafts explanted after the predetermined periods. The pericardial grafts were implanted in 1 animal each for scheduled periods of 3 days, 2 weeks, 1 month, 3 months, and 6 months. The retrieved grafts were processed for light and electron microscopic analyses following gross observation. The glutaraldehyde-treated small-diameter pericardial vascular grafts showed a better endothelialization of the blood contacting surface and a slower fragmentation of the collagen layers than the fresh grafts although it has yet to be proven whether these differences are so significant as to affect the patency results between the groups. PMID- 11872015 TI - The seventh prototype of the mini-spindle-pump: does it fulfill the expectations of a short-term pump? AB - According to recent trends to develop implantable nonpulsatile blood pumps for different function modes and times, our intention was and still is to build a Mini-Spindle-Pump for a pumping duration of about 14 days. Initial conception for this plan was the premise that the device in a mock circuit should move 4 L of water/min at a speed of 12,000 to 15,000 rpm against a pressure difference of 90 mm Hg between pump inlet and outlet. Despite the development of 6 different prototypes, this project was not realized. Under the above-mentioned conditions, the main problem of this type of blood pump, the blood trauma, could not be reduced to an adequate level, i.e., the Mini-Spindle-Pump is not a high speed pump. Therefore, a revision of the conception was necessary. The device in a mock circuit should transport 5 L of water/min at a speed of about 9,000 rpm against a pressure difference of 90 mm Hg between its inlet and outlet. Considering the implantability of the blood pump, the following measurements for its components were arrived at. The U-shaped blockformed plexiglas housing was enlarged to 120 x 40 x 40 mm (length of blood chamber 86 mm, inner diameter 27 mm), and the rotor with 5 windings was redesigned at a length of 64 mm (outer diameter 25 mm, inner diameter 6.7 mm). In a mock circuit, this 7th prototype transported with a speed of 9,000 rpm about 10 L of water/min at an afterload of 80 mm Hg. In acute animal experiments with calves up to 15 h of pumping duration, the device showed the expected efficiency. Experiments with a longer pumping duration are necessary to confirm that this prototype will fulfill the criteria of a short-term pump according to Dr. Y. Nose's advice. PMID- 11872016 TI - Impeller inner diameter in a miniaturized centrifugal blood pump. AB - To design a miniaturized centrifugal blood pump, the impeller internal diameter (ID), which is a circle diameter on the inner edge of the vane, is considered one of the important aspects. Hydraulic performance, hemolysis, and thrombogenicity were evaluated with different impeller IDs. Two impellers were fabricated with an outer diameter of 35 mm, of which 1 had an 8 mm ID impeller and the other had a 12 mm ID. These impellers were combined with 2 different housings in which the inlet port was eccentrically positioned 3.8 and 4.5 mm offset from the center. The hydraulic performance and hemolysis were evaluated in a mock circuit, and thrombogenicity was evaluated in a 2 day ex vivo study with each impeller housing combination. Both impellers required 3,000 rpm in the 3.8 mm offset inlet to attain 5 L/min against 100 mm Hg (left ventricular assist device condition). The 8 mm ID impeller required 3,200 rpm, and the 12 mm ID impeller required 3,100 rpm in the 4.5 mm offset housing. The normalized index of hemolysis was 0.0080 +/- 0.0048 g/100 L in the 8 mm ID impeller with the 3.8 mm offset and 0.022 +/- 0.018 g/100 L with 4.5 mm offset. The 12 mm ID impeller had 0.068 +/- 0.028 g/100 L with the 3.8 mm offset and 0.010 +/- 0.002 g/100 L with the 4.5 mm offset. After the 2 day ex vivo study, no blood clot was formed around the top bearing in all the pump heads. The 8 mm ID impeller with 3.8 mm offset inlet and the 12 mm ID impeller with the 4.5 mm offset had less hemolysis compared to the other pump heads that were subjected to 14 day ex vivo and 10 day in vivo studies. The 8 mm ID impeller with the 3.8 mm offset inlet had a blood clot around the top bearing after the 14 day ex vivo study. No thrombus was found around the top bearing of the 12 mm ID impeller with the 4.5 mm offset in the 10 day in vivo study. These results suggest that the ID does not greatly change the hydraulic performance of a small centrifugal blood pump. The proper combination of the impeller ID and inlet port offset obtains less hemolysis. The larger impeller ID is considered to have less thrombogenicity around the top bearing. PMID- 11872017 TI - Undergraduate urology: a survey of current provisions and guidelines for a core curriculum. PMID- 11872018 TI - Fixation of the testis. PMID- 11872019 TI - Simultaneous bilateral compared with unilateral percutaneous nephrolithotomy. AB - OBJECTIVE: To compare simultaneous bilateral percutaneous nephrolithotomy (SBPCNL) and unilateral PCNL in separate sessions in patients with bilateral renal stones for several variables before and after surgery. PATIENTS AND METHODS: The results from SBPCNL carried out at two centres in different countries on 198 patients (aged 1.25-70 years) were compared with those from 300 patients undergoing unilateral PCNL in separate sessions. At one centre where extracorporeal shockwave lithotripsy (ESWL) was available SBPCNL was used for stones where at least two sessions of ESWL would have been required on each side. Other associated procedures were also used wherever required. RESULTS: The stones were cleared bilaterally in one session of SBPCNL in 190 patients; in eight, fragments of < 3 mm remained but were not clinically significant. A second session was required in six patients on one side only. Variables assessed before and after treatment (e.g. hospital stay, analgesia requirements and complications) were not significantly different between SBPCNL and PCNL. The mean (range) total operative duration for SBPCNL was 46 (20-100) min and the hospital stay 4.3 (3-8) days. CONCLUSION: From this experience, SBPCNL is a cost-effective and beneficial solution for selected patients, with clear advantages over separate unilateral PCNL in patients with bilateral stones. PMID- 11872020 TI - Laparoscopic ureterolithotomy: technical considerations and long-term follow-up. AB - OBJECTIVE: To analyse the technical details and the long-term results of laparoscopic ureterolithotomy. PATIENTS AND METHODS: Laparoscopic ureterolithotomy was undertaken in 101 patients between 1991 and 2001; in only one patient with retroperitoneal fibrous adhesions was the procedure transperitoneal, being retroperitoneal in all the others, using Gaur's balloon retroperitoneoscopy. The mean (range) stone size was 16 (10-47) mm, and the stones were in the upper ureter in 75, mid-ureter in 11 and lower ureter in 15 patients. Nine patients had more than one stone, the maximum being six, in a megaureter. Most were impacted for > 2 months, the maximum being 240 months. RESULTS: Laparoscopic ureterolithotomy was successful in 93 patients, with the eight failures being mostly early in the series. The mean operative duration was 79 min (66 min when the ureter was left open and 92 min when it was sutured). The overall mean duration of urinary leakage was 5.5 days, which was reduced to 3.2 days by stenting and suturing the ureter. The mean (range) blood loss was 25 (5 100) mL. The overall complication rate was high (31%) because of prolonged urinary leakage in 20 patients. No patient required morphine for pain relief and the mean for oral analgesic use was 2.5 days. The mean hospital stay was 3.5 days and that for resuming work 14 (7-28) days. CONCLUSIONS: Laparoscopic ureterolithotomy by the retroperitoneal approach is a safe and reliable minimally invasive procedure. Although its role as a salvage procedure for failed extracorporeal shock wave lithotripsy and ureteroscopy is undisputed, in selected patients with large chronically impacted ureteric stones and particularly with solitary kidneys, it may be considered the first-line treatment. PMID- 11872021 TI - Is sexual intercourse a significant cause of haematuria? AB - OBJECTIVES: To determine whether recent sexual intercourse might be a cause of microscopic haematuria in patients referred to a urological unit following dipstick detection of urinary haemoglobin. SUBJECTS AND METHODS: Forty-eight volunteers (24 men and 24 women) consented to have heterosexual intercourse with their regular partner, and to provide samples of urine for testing before and from the first void on the morning after intercourse. After appropriate instruction, volunteers tested their own urine for the presence of blood using standard dipsticks. Any volunteer with haematuria either before or after intercourse was offered a standard haematuria assessment. The results were analysed using the chi-squared test. RESULTS: None of the volunteers tested positively for haematuria immediately before sexual intercourse; six of the 24 women (25%), but no men, became positive after intercourse (P < 0.01). Only one of the six women accepted the offer of a haematuria evaluation and no pathology was identified. CONCLUSION: These results suggest that up to a quarter of women develop microscopic haematuria as a direct result of sexual intercourse. A history of recent sexual intercourse should therefore be considered when assessing the clinical significance of microscopic haematuria in women. PMID- 11872022 TI - The community-based morbidity of flexible cystoscopy. AB - OBJECTIVE: To evaluate patients' experience after flexible cystoscopy (FC), particularly concentrating on the prevalence and degree of symptoms, the frequency of visits to their General Practitioner (GP), subsequent antibiotic rates and the actual incidence of urinary tract infection (UTI). PATIENTS AND METHODS: Consecutive patients (420) presenting for FC were audited prospectively. A pain score for the procedure was recorded immediately afterward (linear scale 0 10) and a self-administered questionnaire completed at 7 days, to assess the objective and subjective symptoms and their duration, and the incidence of GP visits and subsequent antibiotic provision noted. An interim analysis was conducted on the initial 274 datasets received. To estimate the incidence of FC induced UTI, the final 110 patients were asked not to consult their GP but to present to the urology department at 3 days after FC (or the emergency department if clinically necessary). These patients had initially provided a mid-stream urine (MSU) sample before FC and were assessed symptomatically with a subsequent sample obtained if a urinary dipstick test 3-days after FC was abnormal. RESULTS: In all, 384 (91%) evaluable forms were returned. The median (range) pain score for FC was 1.1 (0-8.5), with seven patients (1.8%) recording a pain score of > 5 (all men); 382 patients (99.5%) declared they would be happy to undergo an identical procedure in the future if medically indicated. Pain on voiding was reported in 190 patients (50%), urinary frequency in 142 (37%) and gross haematuria in 73 (19%). Eighteen of the initial 274 patients (6.6%) visited their GP, with 15 (5.5%) of these receiving antibiotics. The MSU data from the final 110 patients showed a FC-mediated infection in three (2.7%). CONCLUSION: Although FC is well tolerated, gross haematuria, urinary frequency and dysuria occur afterward much more frequently than expected. Patients should be thoroughly counselled before FC about these potential symptoms, to reduce their concern, any unnecessary GP visits and the use of antibiotics. PMID- 11872023 TI - Urgency, urge incontinence and voiding symptoms in men and women aged 70 years and over. AB - OBJECTIVE: To evaluate the prevalence of urgency, urge incontinence and voiding symptoms, and their associations in older men and women. SUBJECTS AND METHODS: A population-based cross-sectional survey was conducted involving 171 men and 227 women aged > or = 70 years. The data were collected by interview, with a response rate of 92.8%. Voiding symptoms were defined as weakened and/or intermittent stream. The prevalence of urgency, urge incontinence and voiding symptoms were calculated for men and women aged 70-79 and > or = 80 years. Logistic regression models were used to examine the association of voiding symptoms with urgency, with or without incontinence, adjusted in the separate models for age and in the combined model also for gender. RESULTS: The prevalence of urge incontinence was higher than urgency alone in both men and women (23.9% vs. 9.8% and 36.4% vs. 8.6%, respectively); 71.9% of the men and 48.3% of the women reported voiding symptoms (P < 0.001). Men and women with voiding symptoms were both significantly more likely to report urgency with or without incontinence than those with no voiding symptoms (odds ratio 3.49, 95% confidence interval 1.42-8.57, and 2.34, 1.31-4.17, respectively). Age had no independent effect in men, and in women the effect was marginal. In the combined model female gender (1.98, 1.25-3.16) increased the risk of urgency with or without incontinence. CONCLUSION: Urgency, urge incontinence and voiding symptoms are common and associated with each other in older men and women; the association is stronger in men. Women are at greater risk of having urgency with or without incontinence. Because the study was cross sectional a causal relationship cannot be confirmed. PMID- 11872024 TI - The cost-effectiveness of preoperative testing (basic office assessment vs. urodynamics) for stress urinary incontinence in women. AB - OBJECTIVE: To compare the cost-effectiveness of preoperative testing strategies in women with stress incontinence symptoms, as although urodynamic testing is used to improve the diagnostic accuracy in women with incontinence, the clinical and economic consequences of different levels of testing have not been evaluated. MATERIALS AND METHODS: Decision analysis was used to evaluate basic office assessment (BOA) and urodynamic testing for women with stress incontinence symptoms who were candidates for primary surgical treatment. Costs were calculated using the Federal Register. Parameter estimates for the effectiveness of treatment for different diagnoses of incontinence were based on published reports. Incremental cost-effectiveness was defined as the cost in dollars per additional patient cured of incontinence. RESULTS: Urodynamics did not improve the effectiveness of treatment; both strategies of a BOA and urodynamic testing resulted in a cure rate of 96% after initial and secondary treatments. The mean cost of care (including initial and secondary treatments and outcomes) was similar for the two strategies ($5042 for BOA, $5046 for urodynamic testing). With BOA reduced testing costs were balanced by increased costs for patients who failed the initial treatment. Under baseline assumptions, one additional cure of incontinence (incremental cost-effectiveness) using the urodynamic strategy cost $3847, compared with BOA. By sensitivity analyses, BOA was less costly than urodynamics when the prevalence of genuine stress incontinence was > or = 80%. CONCLUSION: These findings do not support the routine use of urodynamics before surgery in women likely to have genuine stress incontinence, and provide the justification for randomized trials of preoperative testing strategies. PMID- 11872025 TI - Artificial urinary sphincter implantation in the irradiated patient: safety, efficacy and satisfaction. AB - OBJECTIVE: To compare the long-term outcome of artificial urinary sphincter (AUS) implantation in patients after prostatectomy, with and with no history of previous irradiation. PATIENTS AND METHODS: The study included 98 men (mean age 68 years) with urinary incontinence after prostatectomy for prostate cancer (85 radical, 13 transurethral resection) who had an AUS implanted. Twenty-two of the patients had received adjuvant external beam irradiation before AUS implantation. Over a mean (range) follow-up of 46 (5-118) months, the complication and surgical revision rates were recorded and compared between irradiated and unirradiated patients. The two groups were also compared for the resolution of incontinence and satisfaction, assessed using a questionnaire. RESULTS: Overall, surgical revision was equally common in irradiated (36%) and unirradiated (24%) patients. After activating the AUS, urethral atrophy, infection and erosion requiring surgical revision were more common in irradiated patients (41% vs. 11%; P < 0.05); 70% of patients reported a significant improvement in continence, regardless of previous irradiation. Patient satisfaction remained high, with > 80% of patients stating that they would undergo surgery again and/or recommend it to others, despite previous irradiation and/or the need for surgical revision. CONCLUSIONS: Despite higher complication and surgical revision rates in patients who have an AUS implanted and have a history of previous irradiation, the long term continence and patient satisfaction appear not to be adversely affected. PMID- 11872026 TI - A study comparing various noninvasive methods of detecting bladder cancer in urine. AB - OBJECTIVES: To compare the nuclear matrix protein (NMP)-22 assay, bladder tumour specific antigen (BTAstat) test, telomerase activity (using the telomeric repeat amplification protocol assay, TRAP) and a haemoglobin dipstick test for their ability to replace voided urine cytology (VUC) for detecting bladder cancer. PATIENTS AND METHODS: The study included 120 urological patients prospectively recruited and assessed before surgery. A single freshly voided urine sample (approximate 100 mL) was collected from each patient and aliquoted for each test. All assays were conducted according to the manufactures' guidelines; 79 patients were tested for telomerase activity. The results were then compared with VUC and the diagnosis confirmed by cystoscopy and histology. RESULTS: Fifty-two patients had histologically confirmed transitional cell carcinoma. The overall sensitivity for BTAstat, NMP22, telomerase, VUC and dipstick testing was 63%, 81%, 84%, 48% and 50%, respectively. Combining the results for telomerase and NMP22 gave a sensitivity of 100%. For G1 tumours the respective sensitivities were 23%, 62%, 56%, 23% and 15%, for G2 tumours, 68%, 86%, 92%, 50% and 41% and for G3 tumours 88%, 88%, 100%, 71% and 82%. For pTa tumours the respective detection rates were 48%, 70%, 84%, 39% and 30%, for pT1 tumours 80%, 90%, 90%, 50% and 50%, for pT2/pTis tumours, 100/100%, 100/100%, 100/100%, 88/100% and 88/83%. The overall specificity for the respective tests was 82%, 87%, 93%, 87% and 54%; combining the results of NMP22 and telomerase activity increased the specificity to 96%. CONCLUSIONS: There was significantly better detection than VUC when using the NMP22 and TRAP assay, especially for well-differentiated (P < 0.001 and 0.0027, respectively) and superficial tumours (P < 0.001 and 0.034, respectively). Combining the results of NMP22 and telomerase activity yielded values comparable with cystoscopy. PMID- 11872027 TI - Outcome of radical cystectomy for bladder cancer according to the disease type at presentation. AB - OBJECTIVE: To examine whether the outcome of cystectomy for invasive transitional cell carcinoma (TCC) of the bladder was influenced by the type of disease at initial presentation. PATIENTS AND METHODS: The charts of 76 patients treated for TCC by radical cystectomy from 1987 to 1997 in our unit were reviewed. The patients were divided into three groups: group 1 comprised 43 patients with primary invasive disease; group 2 included 12 patients with progression of an initial superficial bladder tumour after failure of conservative treatment; and group 3 comprised 21 patients who had a radical cystectomy for superficial TCC, with a high risk of progression after attempts at conservative treatment. The pathological findings on transurethral resection and cystectomy specimens, cancer specific survival and the time to progression were compared among the three groups. RESULTS: The rate of pT0 in cystectomy specimens was 16%, 41% and 24% in groups 1, 2 and 3, respectively. Under-staging occurred in 24% of cases in group 3. The 10-year cancer-specific survival rates were 48%, 47% and 82% in groups 1, 2 and 3, respectively. The cancer-specific survival rate and progression rate were not significantly different between groups 1 and 2, but were significantly lower/higher in these patients than in group 3 (P < 0.01). CONCLUSIONS: These data suggest that the prognosis of superficial TCC which progresses despite conservative management is no better than that of invasive TCC at initial presentation, despite the closer follow-up received by the former patients. Early identification of this group of patients may improve the cancer-specific survival, as early cystectomy for high-risk superficial TCC yields better results. PMID- 11872028 TI - Hot flushes and prostate cancer: pathogenesis and treatment. PMID- 11872029 TI - Serendipity in detecting disease in low prostate-specific antigen ranges. AB - OBJECTIVE: To assess the magnitude of prostate cancer detection by serendipity (the coincidental detection of prostate cancer during the evaluation of an abnormal screening test result) when a digital rectal examination (DRE) and transrectal ultrasonography (TRUS) are used as initial screening tests for prostate cancer in men with low levels of prostate-specific antigen (PSA; 0.0-3.9 ng/mL). PATIENTS AND METHODS: In all, 117 participants of a population-based screening study were diagnosed with prostate cancer after a standard evaluation of an abnormal screening test result; 49 underwent radical prostatectomy. Serendipity was defined as either: (i) the presence of prostate cancer opposite to the side that raised suspicion for cancer on DRE and/or TRUS; (ii) a negative lesion-directed biopsy while cancer was present in one or more of the cores of the sextant biopsy; (iii) a tumour volume of < 0.5 mL on radical prostatectomy. RESULTS: Depending on the definition, 27-63% of prostate cancers detected at low PSA values were detected coincidentally and not as a result of a true-positive test result. The proportion of cancers detected by serendipity was inversely correlated with serum PSA level. CONCLUSION: A relatively high proportion of prostate cancers diagnosed in men with low PSA levels, and in which a biopsy was prompted by a suspicious DRE and/or TRUS, are considered to be detected by chance only. As these cancers are mostly small (< 0.5 mL), with potentially low biological aggressiveness, relying on serendipity seems disadvantageous in prostate-cancer screening. The level of serendipity in prostate cancer detection, the poor performance of the screening test, and high inter-observer variability, casts further doubt on the utility of DRE (and TRUS) as initial screening tests for prostate cancer in population-based screening. PMID- 11872030 TI - Clinicopathological features of prostate cancer in Jamaican men. AB - OBJECTIVE: To document the clinicopathological features of prostate cancer in a cohort of Jamaican men, and to determine which of these features are of prognostic significance in this population. PATIENTS AND METHODS: The clinical and pathological findings in 99 patients with prostate cancer (diagnosed consecutively after biopsy, in the Department of Pathology at the University of the West Indies) between 1993 and 1997 were reviewed retrospectively. Biopsy specimens included 74 needle biopsies and 25 transurethral resection (TUR) specimens. RESULTS: The mean age at diagnosis was 72.3 years and 79 patients (80%) were symptomatic. The median (range, interquartile range) serum prostate specific antigen (PSA) value at diagnosis was 37 (1-2100, 2-750) ng/mL; 63% of the patients had clinical stage T1 or T2 disease. Most (60%) of the cancers had a Gleason score of 8-10. Perineural invasion was present in a third of cases overall; high-grade prostatic intraepithelial neoplasia and periprostatic involvement were present in 18% and 8% of biopsies, respectively. The median percentage involvement of all biopsy samples was 37%, that for needle biopsies 47% and for TUR specimens 14%. Of the 90 patients with complete follow-up data, 37 (41%) died; the cause was progressive disease in 19 (51%). The mean (sd, range) survival was 41.3 (19.7, 1-73) months. On univariate analysis, age, PSA level, tumour stage, Gleason score, perineural involvement and periprostatic involvement were significantly associated with an increased risk of dying from prostatic cancer; in a multivariate model, PSA and tumour stage (4 vs. 1) were the only independent factors. CONCLUSIONS: The mean PSA values at the time of diagnosis, the median percentage of biopsy involvement by cancer and the number of patients with tumours of high histological grade were comparatively high, probably reflecting the patients' relatively late clinical presentation. Established prognostic markers were predictive of the risk of death from prostate cancer. PMID- 11872031 TI - The relationship of circulating insulin-like growth factor 1, its binding protein 3, prostate-specific antigen and C-reactive protein with disease stage in prostate cancer. AB - OBJECTIVE: To assess the relationships among circulating insulin-like growth factor-1 (IGF-1), IGF binding protein-3 (IGFBP-3), prostate-specific antigen (PSA) and C-reactive protein (an indicator of inflammatory systemic response) in patients with prostate disease and cancer of different stages. PATIENTS AND METHODS: Circulating IGF-1, IGFBP-3, PSA and C-reactive protein concentrations were measured in patients with BPH (17) or stages T1/T2 (15), T3/T4 (16) and metastatic prostate cancer (12 patients). RESULTS: IGF-1 and IGFBP-3 concentrations were similar between patients with BPH and those with cancer, and there was no difference between the groups with cancer. In the latter patients there was a significant correlation between age and IGFBP-3 concentrations (r = 0.400, P = 0.008) but not with IGF-1 concentrations. Controlling for age, there were significant partial correlations between C-reactive protein and IGF-1 (r = 0.412, P = 0.008) and IGFBP-3 (r = -0.277, P = 0.05). CONCLUSIONS: These results indicate that circulating IGF-1 and IGFBP-3 concentrations do not change with disease stage in prostate cancer, but that they decrease with an increase in the systemic inflammatory response. PMID- 11872032 TI - E-cadherin and beta-catenin are down-regulated in prostatic bone metastases. AB - OBJECTIVE: To determine the E-cadherin and beta-catenin expression phenotype in untreated primary prostate cancer and corresponding bone metastases. MATERIALS AND METHODS: Paired bone metastasis and primary prostate specimens were obtained from 14 men with untreated metastatic prostate carcinoma. The tumours were histologically graded by an independent pathologist. Expression of mRNA for E cadherin and beta-catenin was detected within the tumour cells using in-situ hybridization with a 35S-labelled cDNA probe. The expression of E-cadherin and beta-catenin were graded as uniform, heterogeneous or negative. RESULTS: The mRNA for E-cadherin was expressed in 13 of 14 primary carcinomas and 11 bone metastases; beta-catenin was expressed by 13 and nine, respectively. Of the primary tumours, nine expressed E-cadherin and beta-catenin uniformly; in contrast, all metastases had down-regulated E-cadherin and/or beta-catenin. CONCLUSIONS: The down-regulation of E-cadherin and beta-catenin are a feature of the metastatic phenotype, which may be a significant factor in the genesis of bone metastases. However, this does not appear to be reflected in the expression of these molecules in the primary tumours. PMID- 11872033 TI - The Lue procedure: an analysis of the outcome in Peyronie's disease. AB - OBJECTIVE: To assess the Lue procedure (plaque incision and venous grafting) for correcting the penile deformity of Peyronie's disease (which can cause penile shortening and erectile dysfunction) as an alternative to the Nesbit procedure (which can worsen the shortening). PATIENTS AND METHODS: Fifty-one patients (mean age 51 years, range 27-68) with Peyronie's disease had their penile deformity corrected by plaque incision and saphenous vein grafting. All patients had stable Peyronie's disease and a mean (range) penile deformity of 57 (20-90). The vein graft was taken from the long saphenous vein at the ankle or groin and several sites grafted in 14 patients. The mean follow-up was 16 months. RESULTS: An excellent or satisfactory result was obtained in 47 patients (92%); the penis was completely straightened in 42 (82%). Four patients (8%) developed postoperative erectile dysfunction. Eighteen men (35%) had some degree of penile shortening (> 1 cm in eight), among whom intercourse was affected to a variable extent in six (12%). CONCLUSION: The Lue procedure is an effective option in the surgical management of Peyronie's disease, but penile shortening after surgery remains a risk. PMID- 11872034 TI - A European multicentre study to evaluate the tolerability of apomorphine sublingual administered in a forced dose-escalation regimen in patients with erectile dysfunction. AB - OBJECTIVE: To determine the risk-benefit ratio of a forced dose-escalation regimen (2 to 3 to 4 mg) in a European clinical study evaluating apomorphine sublingual (SL) in treating erectile dysfunction (ED), by evaluating the overall tolerability and efficacy of the regimen compared with placebo in patients with ED, and evaluating efficacy by assessing the proportion of successful attempts resulting in sexual intercourse. PATIENTS AND METHODS: This randomized, double blind, two-arm, parallel-group study was conducted in 507 patients enrolled at 34 European sites. After a 1-2 week screening period, patients were treated for 8 weeks with either placebo or apomorphine SL administered as a forced dose escalation regimen. Heterosexual men (aged 18-70 years) were eligible for participation in the study if they were in stable health, a stable relationship of > or = 6 months duration, had a history of erectile inability, and were diagnosed with ED (successful in fewer than half of attempts to attain and maintain an erection firm enough for intercourse during the 30 days before screening). Patients provided information (recorded on diary cards and reviewed at each study visit) about the frequency and success in achieving erections and of sexual intercourse attempts during both the screening and treatment periods. The dosing regimen required patients to take one tablet of apomorphine SL (2 mg for 2 weeks, then 3 mg for 2 weeks and finally 4 mg for the remaining 4 weeks) or placebo 15-25 min before intercourse, and intercourse was to be attempted at least twice a week. Safety data were collected throughout the 8-week study period, and included recording adverse events, vital signs and changes in laboratory test values for standard haematology and biochemistry variables. The primary efficacy variable was the proportion of successful attempts, defined as an erection rigid enough for sexual intercourse, occurring after dosing (successful intercourse rate). The proportion of erections achieved was a secondary efficacy variable. RESULTS: Of the 507 patients, 254 received apomorphine SL and 253 received placebo; 87% of patients in both groups completed the 8-week treatment period. Of the patients receiving apomorphine SL, 24% had hypertension, 11% had coronary artery disease, 10% had diabetes, and 5.5% had benign prostatic hypertrophy; 62.6% of treated patients received concomitant medications for these maladies. The treatment groups were balanced for demographic and baseline variables, including comorbidity factors. Treatment emergent adverse events, reported by > 5% of patients in the treated group, were nausea (9.8%), dizziness (7.1%) and headache (6.7%), compared with 0.4%, 2.4% and 4.0%, respectively, in the placebo group. Sixty-six patients withdrew from the study, 16 because of study drug-related adverse events (12 from the apomorphine and four from the placebo group). Six patients (three in each group) reported a total of nine serious treatment-emergent adverse events, all of which resolved by the end of the study. In the intention-to-treat population, the proportion of successful attempts at sexual intercourse and of erections were statistically greater in the apomorphine than in the placebo group (P = 0.001 and 0.021, respectively); analysis of the per-protocol population results confirmed this significant difference. CONCLUSION: This European study supports the safety and tolerability of apomorphine SL despite the forced escalation to a 4-mg dose (exceeding the approved 2-3 mg dose). Adverse effects were not treatment limiting. These results further support the clinically significant efficacy of apomorphine SL for treating ED at all doses used. The risk/benefit ratio supports apomorphine SL as a safe and effective alternative in managing ED. PMID- 11872035 TI - The role of pelvic traction in the management of primary monosymptomatic nocturnal enuresis. AB - OBJECTIVE: To determine whether pelvic traction is beneficial in children with primary nocturnal enuresis. PATIENTS AND METHODS: There can be disproportionate growth between the spinal column and neural tube in prepubertal children. The normal elongation of the vertebral column in children during sleep could stretch the filum terminale and nerve roots, representing a minor degree of tethering that affects neural function and contributes to nocturnal enuresis. Pelvic traction induces a similar or more intense stretch while a patient is fully awake (and able to control their bladder). Releasing the potential tethering in this way, combined with conditioning therapy, could be beneficial. Fifty patients (aged 7-17 years) with monosymptomatic primary nocturnal enuresis were evaluated in a prospective study. All had 10 sessions of pelvic traction applied over 4 weeks and were followed up for 3 months afterward; no other medications were given. RESULTS: All patients had fewer wet nights, with variable degrees of success (20-80%) during and 3 months after traction. CONCLUSION: Pelvic traction is a safe, simple, economic and effective treatment for primary monosymptomatic nocturnal enuresis. PMID- 11872036 TI - Experience with the long-term use of desmopressin for nocturnal enuresis in children and adolescents. PMID- 11872037 TI - Knowledge and attitudes of Korean parents towards their son's circumcision: a nationwide questionnaire study. AB - OBJECTIVE: To evaluate knowledge about the foreskin and circumcision, and to understand the attitudes of parents to circumcision in Korea, where circumcision in childhood is widely practised with no particular religious or medical background. SUBJECTS AND METHODS: A nationwide study involving questionnaires was conducted on 5500 parents with at least one son attending elementary school. Responses were obtained from one of the parents. RESULTS: The response rate was 76.1% (4183); circumcision was most common in boys when aged 11 years, followed by neonatal circumcision. Of the parents, 91.3% believed that circumcision is necessary, while 2.1% believed it to be unnecessary. The principal reasons given for circumcision were 'to improve penile hygiene' (82.4%), followed by 'to improve future sexual potency' (7.5%). Among those who did not believe circumcision to be necessary, the most common reason was the expectation of spontaneous retraction of the prepuce with age (55.1%). Most (88.4%) of the parents believed that smegma is not a clean material, and is infected by microorganisms. Most parents (80.6%) thought that circumcision would prevent genital tract infection of the future spouse. Peer pressure was one of the most influential factors in deciding upon circumcision; 41.9% of the parents were anxious that their child might be ridiculed by his peer group unless he was circumcised, while 27.4% of the parents believed that their child might be ridiculed if he was circumcised. Mothers were more positive about circumcision than fathers (P < 0.05). Parents with a higher education and higher socio economic status were also more positive about circumcision (P < 0.05). Mothers were prone to emphasize improved sexual potency (P < 0.05). There was no significant difference in response between urban and rural areas. CONCLUSION: This study indicates that common beliefs held by parents about the prepuce or circumcision differ significantly from current medical knowledge, and these beliefs have a major influence on the practice of circumcision in Korea. More clinical research on the natural history of the foreskin is needed, and it is critical that both children and parents are informed about the potential benefits and disadvantages of circumcision. PMID- 11872038 TI - Influence of the bridging effect by sequential laser application on tissue ablation in an ex vivo model, taking organ perfusion into account. AB - OBJECTIVE: To improve the efficacy of interstitial laser coagulation of tissue by causing a 'bridging' effect, using a sequential multiple-probe procedure on an ex vivo kidney model, as only a limited area of tissue is destroyed with a single probe and the coagulation takes longer when multiple punctures are used. MATERIALS AND METHODS: A laser beam was generated using a Nd:YAG laser and applied to kidney tissue by a diffuser tip (quartz glass cap 2 x 19 mm). For sequential laser application (SLA), three probes were placed through punctures into kidney tissue, at 5 or 10 mm apart. The laser energy was applied in different time-energy combinations through the three probes. The effect of SLA was compared with that from one probe delivering the optimal PowerMode 180 protocol (Dornier, Germering, Germany), which represents the standard energy protocol for the clinical treatment of parenchymal organs using this laser device. An isolated porcine kidney was chosen for laser coagulation under different conditions of perfusion. The ablative efficacy was defined as the volume of necrosis per minute. RESULTS: Applying various time-energy combinations to isolated unperfused porcine kidney caused extensive tissue ablation (5.6 mL). In trials with saline and blood perfusion for improved cooling, the necrotic volume was 2.5 and 3.9 mL, respectively (with no carbonization, 3.2 mL). Compared with a single-probe procedure, the ablation efficacy was 10 times better with SLA coagulation. This improvement was initiated by the bridging effect: coagulation in neighbouring areas affects perfusion and convection to an extent that induces the formation of bridges of necrosis between the probes. CONCLUSION: Tissue ablation is markedly improved by interstitial laser coagulation using a sequential multiple-probe technique. PMID- 11872039 TI - Effects of electropermeabilization after the administration of anticancer drugs on transitional cell carcinoma. AB - OBJECTIVE: To assess in vitro and in vivo the potential utility of electropermeabilization (EP) as an anticancer drug delivery system for the treatment of transitional cell carcinoma (TCC). MATERIALS AND METHODS: To analyse the effects of EP on the internalization of adriamycin and bleomycin by cells, aliquots of a suspension of YTS-1 carcinoma cells (derived from a human TCC line) were mixed with a solution of adriamycin or bleomycin and then exposed immediately to an electric field (1000 V/cm, 1 Hz, 100-micros square wave, eight pulses). After a 2-h incubation the concentration of each drug in the cells was measured by high-performance liquid chromatography (for adriamycin) and by bioassay (for bleomycin). The concentrations of drugs in the cells were compared with untreated cells. To analyse the effects of EP on cytotoxicity, the same treatments were applied to a suspension of cells plus adriamycin or bleomycin and then the cells incubated for 6 h with tritiated thymidine ([3H]-TdR), monitoring the incorporation of [3H]-TdR into the cells. Cells with no electrotreatment acted as controls. To assess the effects on tumours in vivo, YTS-1 cells were transplanted subcutaneously into nude mice; when the tumours had reached 7 mm in diameter, EP was applied (using electrical pulses as before, 10 min after the direct injection of bleomycin into the tumour). Tumours were then measured regularly and compared with sham-treated tumours, tumours treated with electric pulses alone, and tumours treated with bleomycin alone. Survival was also compared. RESULTS: There were no significant differences in the levels of adriamycin between cells with and with no EP, whereas there was a marked difference for bleomycin. Growth inhibition by adriamycin with and with no EP was similar, while the growth-inhibitory effects of bleomycin were almost doubled. There was a reduction in tumour size only in the group treated with bleomycin plus electric pulses and two of five tumours disappeared completely. Survival in this group was also significantly better than in the other groups. CONCLUSION: EP after administering bleomycin might be an effective treatment for TCC. PMID- 11872040 TI - Resistive index: an experimental study of the effects of diuretic stimulation in the unobstructed porcine kidney. AB - OBJECTIVE: To study the effects of diuretic stimulation with frusemide and mannitol on the renal resistive index (RI) of the undilated unobstructed porcine kidney. MATERIALS AND METHODS: Experiments were carried out on 14 pigs divided into two equal groups (A and B), under general anaesthesia. In all pigs both ureters were catheterized, thereby allowing urine output to be measured on each side separately, and an indwelling catheter in a femoral artery used to continuously measure the systemic blood pressure. Furthermore, in five pigs from each group, right renal blood flow was measured by ultrasonic flow probes placed around the renal arteries. The right kidney RI was measured at baseline and at 5 min intervals after frusemide (group A) and mannitol (group B) stimulation, for 75 min. RESULTS: Frusemide caused significant increases in diuresis (P < 0.001) and renal blood flow (P = 0.009). This was accompanied by a significant decrease in mean arterial pressure (P < 0.001). The RI was unaffected (P = 0.706), with mean values in group A of 0.58-0.65. Mannitol also caused a significant increase in diuresis (P < 0.001) and a subtle but significant decrease in mean arterial pressure (P < 0.001). However, renal blood flow was unaffected by the diuretic (P = 0.820); the mean RI values were unaffected, at 0.61-0.66 (P = 0.375). CONCLUSIONS: These results show that diuretic stimulation with frusemide or mannitol does not affect the RI of the unobstructed undilated porcine kidney, despite significant changes in diuresis, renal blood flow and mean arterial pressure. PMID- 11872041 TI - The rho/rho-kinase pathway is involved in the progression of testicular germ cell tumour. AB - OBJECTIVE: To clarify the role of one of the downstream effectors of Rho (Rho kinase) in testicular germ cell tumour (GCT) by quantifying mRNA expression for Rho-kinase in patients with this disease. MATERIALS AND METHODS: The mRNA levels of the RhoA and Rho-kinase genes were analysed in surgical specimens of testicular GCT tissues from 57 consecutive Japanese patients, and in the corresponding non-tumour tissue originating from the same patient, using the polymerase chain reaction after reverse transcription. The expression levels of these genes were compared between the tissues and the relationship between their expression levels evaluated within tumours and with tumour stage. The difference in the expression levels of the mRNAs of RhoA and Rho-kinase genes were also assessed between tumours that were seminoma only and mixed tumours of seminoma and nonseminoma. RESULTS: RhoA and Rho-kinase mRNAs were more abundant in tumour tissue than in non-tumour tissue (P < 0.01 and < 0.05, respectively). High RhoA and Rho-kinase mRNA expressions were related to tumour stage (P < 0.05 and < 0.01, respectively). The mRNA levels of RhoA and Rho-kinase in mixed tumours were higher than in tumours with seminoma only (P < 0.01 and < 0.05, respectively). There was a positive relationship between expression levels of mRNAs of RhoA and Rho-kinase in tumour tissues (P < 0.001). CONCLUSIONS: These findings suggest that the RhoA/Rho-kinase pathway is involved in the progression of testicular GCT. This pathway might be a molecular target for new treatment strategies for this disease. PMID- 11872042 TI - Cisplatin-induced in vivo differentiation of human embryonal carcinoma. AB - OBJECTIVE: To investigate the differentiation of embryonal carcinoma (EC) by cisplatin, and the underlying mechanism, as untreated metastases of nonseminomatous germ cell tumours rarely consist of fully differentiated mature somatic tissues, but such mature metastases are more common after various treatments, including chemotherapy. MATERIALS AND METHODS: The TTSC-3 human testicular EC line heterotransplanted into nude mice was used as a target. After treating tumour-bearing mice with intraperitoneal injections of varying doses of cisplatin, the histopathology of the tumours was assessed and various gene expressions in the tumours determined by cDNA-array technology. RESULTS: When cisplatin at 1 mg/kg/week was injected intraperitoneally into TTSC-3-bearing mice, there was no effect on tumour growth. However, injecting cisplatin at 5 mg/kg/week induced a marked regression of the tumour. In contrast, cisplatin at 3 mg/kg/week had a modest inhibitory effect on tumour growth and induced tumour dormancy. Histological examination showed that 5 weeks after injecting cisplatin (3 mg/kg/week), primitive mesenchymal-like cells increased, and 10 weeks afterward cartilage and well-developed glands (teratoma) were apparent; at > 15 weeks afterward there were no EC cells visible. cDNA probes from reverse transcribed mRNAs of TTSC-3 treated with cisplatin or saline for 10 weeks were compared to identify genes differentially expressed in cisplatin-treated TTSC-3. Of 1176 different human cDNA transcripts in cisplatin-treated TTSC-3, three genes (tumour necrosis factor receptor 1, caspase 8 and Apaf1), which are associated with apoptosis, were expressed markedly more than after saline injection. CONCLUSIONS: The intermediate dose of cisplatin inhibited tumour growth of EC by inducing differentiation and enhancing apoptosis-related gene expression. These findings suggest that cisplatin may play a significant role in the differentiation of EC in vivo. PMID- 11872043 TI - Substitution of the distal female urethra with a vaginal flap and pedicled skin island. PMID- 11872044 TI - Appendicular abscess presenting as an infiltrating bladder tumour. PMID- 11872045 TI - Penile preservation in squamous cell carcinoma of the bulbomembranous urethra. PMID- 11872047 TI - Treatment of axillary hyperhidrosis. PMID- 11872048 TI - Anti-human epidermal growth factor receptor 2 monoclonal antibody therapy for breast cancer. AB - BACKGROUND: Advances in molecular biology and improved understanding of tumour biology have led to the development of novel treatments for cancer. Trastuzumab (Herceptin; Genentech, San Francisco, California, USA) is a monoclonal antibody directed against human epidermal growth factor receptor (HER) 2 protein, which is overexpressed in a wide variety of human cancers, including 20-30 per cent of human breast cancers. HER-2 plays an important role in oncogenic transformation, tumorigenesis and metastatic spread. Overexpression is associated with a poor prognosis and predicts a poor response to several treatment modalities. METHOD: Literature relating to the monoclonal antibody was identified by a Medline literature search and by cross-referencing from the references of seminal articles on the subject. Four major clinical trials were identified and reviewed. RESULTS AND CONCLUSION: In clinical trials approximately 15-20 per cent of patients with HER-2-overexpressing tumours benefited from treatment with trastuzumab. In sensitive patients the antibody appeared to have intrinsic anticancer activity when given as a single agent. In combination chemotherapy it appeared to act synergistically with other agents. Ongoing research is evaluating trastuzumab in combination with numerous standard chemotherapy regimens and with other novel chemotherapeutic agents. Clinical trials have also revealed several serious side-effects of monoclonal antibody therapy. Most notable is an unpredictable cardiotoxicity, especially when used in combination with anthracycline-based chemotherapy regimens. PMID- 11872049 TI - Early detection of colorectal cancer using high-magnification chromoscopic colonoscopy. AB - BACKGROUND: Endoscopic techniques aimed at early detection of colorectal cancer (CRC) and its precursors, permitting targeted in vivo luminal treatments, have been developed by the Japanese since the early 1990s. The introduction of this new technology to the UK (i.e. magnification endoscopes) may permit earlier and more accurate diagnosis. According to Japanese data, magnification chromoscopy can be used to predict histology and invasive depth of cancer, and help in the detection of flat and depressed colonic lesions. Flat and depressed lesions are not purely Japanese phenomena: they exist with a similar incidence in the UK. METHODS: A Medline search was performed for the years 1955-2001 using the following medical subject headings and search methodology: colorectal cancer and colonoscopy or aberrant crypt foci or molecular kinetics or flat/depressed lesions or chromoscopy. RESULTS AND CONCLUSION: Early CRC, in the form of flat or depressed lesions, can be difficult to detect using conventional colonoscopic techniques and penetrate the colonic mucosa deeply. The implications of detecting these lesions in relation to current approaches to the prevention of CRC are profound. PMID- 11872050 TI - Randomized clinical trial of screening for abdominal aortic aneurysm in women. AB - BACKGROUND: Screening for abdominal aortic aneurysm (AAA) is commonly restricted to men. Recent studies have indicated a possible increase in deaths due to ruptured AAA in women, and a higher rate of rupture in women than in men. The present report details results from a randomized controlled trial that assessed the effects of screening women for AAA. METHODS: Some 9342 women aged 65-80 years were entered into the trial and randomized to age-matched screen and control groups. A single ultrasonographic scan was offered to women in the screening arm of the study. Women with an AAA received follow-up scans, and were considered for elective surgery if certain criteria were met. RESULTS: The prevalence of AAA was six times lower in women (1.3 per cent) than in men (7.6 per cent). Over 5- and 10-year follow-up intervals, the incidence of rupture was the same in the screened and control groups of women. CONCLUSION: Screening women for AAA is neither clinically indicated nor economically viable. PMID- 11872051 TI - Randomized clinical trial of no wound drains and early discharge in the treatment of women with breast cancer. AB - BACKGROUND: Women undergoing surgery for primary breast cancer routinely have suction drains inserted deep to the wounds, which are removed approximately 6-8 days after operation, requiring a period of stay of that duration in hospital. The aim of this study was to perform a prospective randomized clinical trial to evaluate a new surgical technique of suturing flaps without wound drainage, combined with early discharge, in women undergoing surgery for breast cancer. METHODS: A total of 375 patients undergoing surgery for breast cancer were randomized to conventional surgery or suturing of flaps with no drain. The main outcome measures were length of hospital stay, surgical morbidity, psychological morbidity and health economics. RESULTS: Suturing of flaps and avoiding wound drainage in women undergoing surgery for breast cancer resulted in a significantly shorter hospital stay. Adopting this surgical technique with early discharge did not lead to any difference in surgical or psychological morbidity. Health economic benefits to the National Health Service resulted from saved bed days with no impact on community costs. CONCLUSION: Wound drainage following surgery for breast cancer can be avoided, thereby facilitating early discharge with no associated increase in surgical or psychological morbidity. PMID- 11872052 TI - Randomized clinical trial of non-mesh versus mesh repair of primary inguinal hernia. AB - BACKGROUND: The optimum method for inguinal hernia repair has not yet been determined. The recurrence rate for non-mesh methods varies between 0.2 and 33 per cent. The value of tension-free repair with prosthetic mesh remains to be confirmed. The aim of this study was to compare mesh and non-mesh suture repair of primary inguinal hernias with respect to clinical outcome, quality of life and cost in a multicentre randomized trial in general hospitals. METHODS: Between September 1993 and January 1996, all patients scheduled for repair of a unilateral primary inguinal hernia were randomized to non-mesh or mesh repair. The patients were followed up at 1 week and at 1, 6, 12, 18, 24 and 36 months. Clinical outcome, quality of life and costs were registered. RESULTS: Three hundred patients were randomized of whom 11 were excluded. Three-year recurrence rates differed significantly: 7 per cent for non-mesh repair (n = 143) and 1 per cent for mesh repair (n = 146) (P = 0.009). There were no differences in clinical variables, quality of life and costs. CONCLUSION: Mesh repair of primary inguinal hernia repair is superior to non-mesh repair with regard to hernia recurrence and is cost-effective. Postoperative complications, pain and quality of life did not differ between groups. PMID- 11872053 TI - Dynamic nature of early organ dysfunction determines outcome in acute pancreatitis. AB - BACKGROUND: All patients with organ dysfunction are currently classified as having severe acute pancreatitis. The aim of this study was to characterize the systemic inflammatory response syndrome (SIRS) and early organ dysfunction in patients with acute pancreatitis and the relationship with overall mortality. METHODS: Patients with predicted severe acute pancreatitis of less than 48 h duration had daily organ dysfunction scores and SIRS criteria calculated. These features were then correlated with outcome. RESULTS: Of 121 patients, 68 (56 per cent) did not develop organ dysfunction; only two of these patients died (mortality rate 3 per cent). Fifty-three (44 per cent) had early organ dysfunction, of whom 11 died (21 per cent). Organ dysfunction and persistent SIRS were both associated with an increased mortality rate, but on multivariate analysis only deteriorating organ dysfunction was an independent determinant of survival. CONCLUSION: Early organ dysfunction in acute pancreatitis usually resolves and in itself has no significant influence on mortality. In contrast, worsening organ dysfunction was associated with death in more than half of the patients (11 of 20); it is this group of patients who should be classified as having severe acute pancreatitis. PMID- 11872055 TI - Estimation of prognosis after hepatectomy for hepatocellular carcinoma. AB - BACKGROUND: The preferred means of treatment for hepatocellular carcinoma is surgical resection. However, the tumour recurrence rate is high. Accurate estimation of the risk of tumour recurrence after hepatectomy may facilitate the administration of adjuvant therapy after hepatectomy to patients with a high likelihood of tumour recurrence. METHODS: The clinical and pathological profiles of 176 patients undergoing hepatectomy for hepatocellular carcinoma from March 1992 to August 1998 were reviewed. The Kaplan--Meier method and log rank test were used to analyse univariate prognostic factors. The Cox proportional hazard model was used for multivariate analysis. Disease-free and overall cumulative survival rates were estimated with respect to the number of prognostic factors. RESULTS: Independent factors associated with a lower disease-free survival included the presence of venous infiltration, presence of daughter tumours, absence of tumour encapsulation and tumour size exceeding 5 cm. Factors decreasing the overall survival rate included the presence of venous infiltration, absence of tumour encapsulation and surgical resection margin less than 1 cm. The 1-year disease-free survival rate decreased from 77.5(s.e. 5.6) to 14.0(8.5) per cent when the number of risk factors present increased from zero to three. The 5-year survival rate decreased from 60.2(11.7) per cent to zero when the number of risk factors increased from zero to three. CONCLUSION: The deterioration of disease-free or overall survival of patients with hepatocellular carcinoma after hepatectomy correlates with increasing number of risk factors. The number of risk factors can be employed to accurately estimate disease-free and overall survival. PMID- 11872054 TI - Phase I study of percutaneous cryotherapy for colorectal liver metastasis. AB - BACKGROUND: The aim was to determine the safety and feasibility of percutaneous cryotherapy for treating irresectable colorectal liver metastases. METHODS: Liquid nitrogen cryoprobes were inserted percutaneously into metastases using the Seldinger technique under computed tomographic guidance. Single-probe treatments were performed with either 3.6- or 6.3-mm cryoprobes (ice-ball volumes 18 and 59 cm3 respectively), or dual-probe treatments with two adjacent 6.3-mm probes (ice ball volume 205 cm3). Treatment involved a single freeze--thaw cycle. RESULTS: Fifteen patients received 25 single-probe treatments and seven patients received 14 dual-probe treatments. The treatment-related mortality rate was zero and complications occurred after six of 39 treatments. Liver metastasis growth was significantly delayed for 2 months after dual-probe but not single-probe treatment. Metastasis cryotherapy stimulated an immediate rise, followed by a fall, in serum carcinoembryonic antigen (CEA) level, associated with immune upregulation that was significantly greater after dual-probe treatments. CONCLUSION: Ablation zones that were approximately four times larger than those produced by previously described percutaneous techniques delayed the growth of metastases, reduced serum CEA concentration, and induced detectable inflammatory and T-lymphocyte responses. Percutaneous cryotherapy for treatment of colorectal liver metastases is feasible and may have a place in conjunction with chemotherapy. PMID- 11872056 TI - Right-lobe live donor liver transplantation improves survival of patients with acute liver failure. AB - BACKGROUND: Right-lobe live donor liver transplantation (LDLT) is used by many liver transplant centres for treating adult patients with terminal liver disease, but its incremental benefit for the intended recipient over cadaveric liver graft transplantation has not been determined. The impact of LDLT as a proactive approach on the outcome of patients with acute liver failure was analysed. METHODS: From January 1999 to March 2001, right-lobe LDLT was offered proactively to 50 consecutive patients with acute liver failure and their families. The outcome of those who opted for right-lobe LDLT (n = 34) was compared with that of those who did not opt for LDLT (n = 16). RESULTS: In the group that opted for right-lobe LDLT, 16 patients eventually received a live donor right-lobe graft (14 patients survived) and three patients received a cadaveric liver graft that became available while the potential live donor was undergoing evaluation (all three patients survived). Among the group who did not opt for LDLT, only one patient received a cadaveric liver graft and survived. The former group had a higher overall survival rate (17 of 34 versus one of 16). With a proactive approach, the overall transplant rate was increased from four of 50 to 20 of 50. The morbidity rate among donors was low and none died. CONCLUSION: Right-lobe LDLT improves the overall survival rate of patients with acute liver failure and should be considered as one of the treatment options for adult patients with acute liver failure. PMID- 11872057 TI - Incidence and pattern of long saphenous vein duplication and its possible implications for recurrence after varicose vein surgery. AB - BACKGROUND: The incidence of long saphenous vein (LSV) duplication has not been clearly established. This anomaly could have implications for recurrence after varicose vein surgery. METHODS: Some 103 saphenograms obtained in 85 patients being considered for peripheral arterial bypass surgery were reviewed. Non-ionic contrast medium was injected directly into the vein or its tributaries at the ankle. Duplications of the LSV and their relation to thigh and calf perforator veins were assessed and recorded by two independent observers. RESULTS: There was evidence of duplication of the LSV in 50 (49 per cent) of the 103 saphenograms. Most duplications were present in the thigh (88 per cent) and the most common pattern was a closed loop (54 per cent). Perforator veins were connected to one branch of the duplication in 42 per cent of the legs (20 per cent of all 103 legs); in half the perforator vein was connected to the non-dominant branch of the duplication. Only ten of the 18 patients who had bilateral saphenograms had duplications in both legs, and only one patient had the same pattern of duplication on both sides. CONCLUSION: The incidence of LSV duplications is higher than previously reported. PMID- 11872058 TI - Prognostic significance of the circumferential resection margin following total mesorectal excision for rectal cancer. AB - BACKGROUND: Knowledge of prognostic factors following resection of rectal cancer may be used in the selection of patients for adjuvant therapy. This study examined the prognostic impact of the circumferential resection margin on local recurrence, distant metastasis and survival rates. METHODS: A national population based rectal cancer registry included all 3319 new patients from November 1993 to August 1997. Some 686 patients underwent total mesorectal excision with a known circumferential margin. This shortest radial resection margin was measured in fixed specimens. None of the patients had adjuvant radiotherapy. RESULTS: Following potentially curative resection and after a median follow-up of 29 (range 14--60) months, the overall local recurrence rate was 7 per cent (46 of 686 patients): 22 per cent among patients with a positive resection margin and 5 per cent in those with a negative margin (margin greater than 1 mm). Forty per cent of patients with a positive margin developed distant metastasis, compared with 12 per cent of those with a negative margin. With decreasing circumferential margin there was an exponential increase in the rates of local recurrence, metastasis and death. CONCLUSION: The circumferential margin has a significant and major prognostic impact on the rates of local recurrence, distant metastasis and survival. Information on circumferential margin is important in the selection of patients for postoperative adjuvant therapy. PMID- 11872059 TI - Breast reconstruction in the United Kingdom and Ireland. AB - BACKGROUND: Although it is becoming more common, previous surveys have identified concerns regarding the safety of immediate reconstruction following mastectomy. The aims of this study were to define current practice of breast reconstruction in the UK and Ireland, and to identify the characteristics of surgeons who use immediate breast reconstruction. METHODS: : A postal questionnaire survey of 498 consultant breast surgeons in the UK and Ireland was performed in January 2000. RESULTS: There were 376 responses (response rate 76 per cent). Eighty-eight per cent of surgeons 'always' or 'usually' discuss reconstruction with patients due to undergo mastectomy; clinicians with a heavy caseload were significantly more likely to discuss it (odds ratio (OR) 18.45 (95 per cent confidence interval 1.99 to 171.07)). The majority of respondents (57 per cent) preferred delayed to immediate breast reconstruction; 70 per cent believed that immediate reconstruction has disadvantages, most commonly that it interferes with adjuvant therapy (56 per cent). Older surgeons were significantly less likely to perform immediate reconstruction (OR 5.18 (2.21 to 12.11)), and were significantly more likely to believe that immediate breast reconstruction has disadvantages (OR 2.02 (1.01 to 4.05)). Surgeons from Ireland were less likely to discuss and perform breast reconstruction (OR 0.20 (0.10 to 0.43) and 0.27 (0.12 to 0.60) respectively), or to have access to a plastic surgeon (OR 0.22 (0.11 to 0.44)). CONCLUSION: : Significant variation exists in the delivery of breast reconstruction after mastectomy in the UK and Ireland. The age, workload and personal characteristics of the surgeon are important in determining reconstructive practice. PMID- 11872060 TI - Late results of selective axillary surgery based on contact cytology in women with operable breast cancer. AB - BACKGROUND: Interest in the possibility of intraoperative analysis of sentinel lymph nodes to select patients with operable breast cancer for immediate axillary clearance encouraged this review of a long-term experience of selective axillary surgery based on intraoperative contact cytology of conventionally sampled nodes. Survival was assessed as a potential marker for understaging. METHODS: Records of 437 patients who had surgery between 1991 and 1994 were reviewed to compare rates of axillary recurrence in patients who had contact cytology only with those who had contact cytology and axillary clearance. RESULTS: Axillary recurrence occurred in seven (3 per cent) of 219 patients who had negative contact cytology, three (4 per cent) of 75 patients who had positive contact cytology with axillary clearance and one (1 per cent) of 93 who had axillary clearance alone. In patients with positive contact cytology, 131 (78 per cent) of 168 positive nodes were in the sample specimen, which included all positive nodes on 19 occasions. Survival probability at 36, 72 and 96 months was 92, 87 and 84 per cent respectively for patients with negative contact cytology, and 85, 73 and 71 per cent for patients with positive cytology and axillary clearance. CONCLUSION: A selective approach to axillary surgery based on intraoperative contact cytology of sampled lymph nodes gave good long-term control of axillary disease. PMID- 11872061 TI - Surgical workload and outcome after resection for carcinoma of the oesophagus and cardia. AB - BACKGROUND: Performing cancer surgery in high-volume centres may lead to improved outcomes. This study explored the relationship between annual workload and outcome following resection for carcinoma of the oesophagus and cardia. METHODS: The study was a retrospective case-note review of 1125 patients who had surgery for cardio-oesophageal cancer in the West Midlands region of England. Outcome measures were 30-day mortality and long-term survival. RESULTS: The overall 30 day mortality rate was 10.0 per cent with a median survival of 14 months and a 5 year survival rate of 17.2 per cent. Increasing age, advanced stage of disease and emergency resection independently affected outcome adversely. Forty-one infrequent operators (fewer than four resections per year) performed 146 resections (13.0 per cent), 18 intermediate operators (between four and 11 resections per year) performed 488 resections (43.4 per cent) and five frequent operators (12 or more resections per year) performed 491 resections (43.6 per cent). The 30-day mortality rate was greatest in the infrequent group (15.1 per cent) compared with both the intermediate group (6.6 per cent; adjusted odds 0.40, P = 0.004) and the frequent group (11.8 per cent; odds 0.73, P = 0.28). There were no differences in survival rates between the groups, and no difference in outcome between high- and low-volume hospitals. CONCLUSION: In this unselected population-based series there was little evidence of a trend of improving 30-day mortality rate with increasing workload, or between workload and long-term survival. PMID- 11872062 TI - Expandable metallic stents for palliation of malignant pyloric and duodenal obstruction. PMID- 11872063 TI - Ultrasonography is superior to plain radiography in the diagnosis of pneumoperitoneum. AB - BACKGROUND: The aim of this study was to compare plain radiography with abdominal ultrasonography in the detection of pneumoperitoneum. METHODS: A total of 188 patients with suspected hollow organ perforation were studied. All patients had abdominal ultrasonography, upright chest radiography and left lateral decubitus abdominal radiography examinations. The sensitivity, specificity, positive and negative predictive value, and accuracy of chest and abdominal radiography were compared with that of abdominal ultrasonography. RESULTS: One hundred and seventy eight patients underwent laparotomy; 170 patients had hollow organ perforation, five patients had perforated appendicitis and three had acute cholecystitis. In the diagnosis of pneumoperitoneum, ultrasonography had improved sensitivity (92 versus 78 per cent), negative predictive value (39 versus 20 per cent) and accuracy (88 versus 76 per cent), and similar specificity (both 53 per cent) and positive predictive value (95 versus 94 per cent) compared with plain radiography. CONCLUSION: Ultrasonography is more sensitive than plain radiography in the diagnosis of pneumoperitoneum. PMID- 11872064 TI - Treatment and outcome of pancreatic venous graft thrombosis after kidney- pancreas transplantation. AB - BACKGROUND: Pancreas venous graft thrombosis after transplantation is the main non-immunological cause of graft failure and usually results in pancreatectomy. Duplex Doppler ultrasonography is the primary imaging technique for monitoring vascular patency after pancreas transplantation. This study reports the results of rescue treatments for pancreas graft thrombosis after simultaneous pancreas- kidney transplantation. METHODS: One hundred and ninety-six patients with insulin dependent diabetes mellitus received a simultaneous pancreas--kidney transplantation. Venous graft thrombosis was diagnosed in 25 of these patients based on Doppler ultrasonographic findings. RESULTS: Total venous graft thrombosis was diagnosed in 20 symptomatic patients, of whom 14 required graft pancreatectomy. Surgical thrombectomy was attempted in six patients with preserved arterial supply and was successful in four. Partial venous graft thrombosis was diagnosed in five asymptomatic patients; one also had partial splenic artery thrombosis. Rescue graft procedures included systemic anticoagulation (one patient), arterial thrombolysis (one) and venous thrombolysis and/or mechanical venous thrombectomy (four episodes in three patients). Graft rescue was achieved in three patients treated by venous thrombolysis/thrombectomy. CONCLUSION: Doppler ultrasonography allows the appropriate selection of rescue treatment based on the findings of total or partial thrombosis. PMID- 11872065 TI - Postoperative enterococcal infection after treatment of complicated intra abdominal sepsis. AB - BACKGROUND: The prevalence of enterococcal isolation and factors associated with postoperative enterococcal infection remain ill defined. METHODS: A prospective longitudinal observational study was conducted of consecutive patients with a first episode of intra-abdominal infection and a positive microbiological culture who did or did not develop a postoperative septic complication involving enterococci. The prevalence of initial enterococcal isolation was determined for each focus of infection. Postoperative enterococcal infections were related to whether appropriate (piperacillin--tazobactam), suboptimal (carbapenems) or inappropriate (cefotaxime plus metronidazole) antienterococcal therapy had been administered empirically. RESULTS: Enterococci were isolated in 42 (21 per cent) of the 200 patients investigated. The isolation rates were 11 per cent for community-acquired peritonitis, 50 per cent for postoperative peritonitis and 23 per cent for intra-abdominal abscesses of both origins. No enterococci were isolated from 49 patients with perforated appendicitis. Independent factors for postoperative enterococcal infection were type of intra-abdominal infection (P = 0.006), Acute Physiology And Chronic Health Evaluation (APACHE) II score greater than 12 (P = 0.04) and inappropriate empirical antibiotic cover (P = 0.05). Postoperative enterococcal infections were associated with a high mortality rate (21 versus 4 per cent; P < 0.0007). CONCLUSION: Enterococci are frequently isolated from intra-abdominal infections of non-appendiceal origin and are often involved in postoperative infectious complications, particularly peritonitis. Empirical antibiotic therapy covering Enterococcus faecalis should be contemplated in some circumstances. PMID- 11872067 TI - Explanatory models in the interpretations of clinical features of dental patients within a university dental education setting. AB - Clinicians may acquire biased perceptions during their dental education that can affect decisions about treatment/management of dental decay. This study established explanatory models used by students to interpret clinical features of patients. It employed a stereotypical dental patient under standardised consultation conditions to identify the interpretation of oral health/disease features in the eyes of student clinicians. The study aimed to establish the perceptions of the patient as a client of the university dental clinic, as seen through the ideological lens of a formal Dental Education system. The discourse during simulated clinical consultations was qualitatively analysed to interpret values and concepts relevant to the assessment of restorative treatment needs and oral health status. Three constructs during the consultation were identified: the Dual Therapeutic Realms, the Choices Underlying Treatment Options, and the High Risk Triad. Comparing these discourse components, the Patient Factors of the Bader and Shugars model for treatment decisions supported the existence of a core set of themes. It was concluded that certain consultation circumstances influenced the adequacy of diagnostic strategies, mainly by introducing loosely defined but highly specific socio-cultural biases ingrained in the Dental Education concepts and diagnostic/treatment needs systems. PMID- 11872068 TI - Practice-based learning: emerging professional characteristics, self-concepts and patterns of knowing in dental training. AB - This paper reports on a content analysis of a reflective learning activity named "A Learning Experience". The study aimed to elucidate the reflections of dental undergraduates and dental therapy students with the purpose of analysing the meaning they attribute to their experiences in training. Thirty-one student completed worksheets were analysed by drawing themes and patterns from the text. In addition, responses were categorised according to patterns of knowing: empirical, aesthetic, personal and ethical. Findings show that students focused mainly on negative experiences and problem situations. Student reflections showed all 4 patterns of knowing and highlighted emerging professional characteristics and students' own professional self-concepts. Findings are discussed in the light of a growing trend in higher and professional education to encourage reflection on learning and practice. PMID- 11872069 TI - Self-assessed clinical competence: a comparison between students in an advanced dental education elective and in the general clinic. AB - A six-month final-year elective, based on an advanced education in general dentistry (AEGD) model, was conducted in an AEGD Clinic to enable dental students to treat patients in a generalist comprehensive setting. The purpose of this study was to evaluate the elective with regard to students' self-assessed clinical skills, as self-assessment is recognized as a means to maintaining competency. Pre- and post-experience surveys were administered to 'AEGD elective students' (n=13) and a comparison group of 'control dental students' (n=18) under a discipline-based clinical curriculum. The survey instrument consisted of 75 self-rated statements on a wide range of clinical and patient management skills. Pre-experience means were not statistically different. The AEGD elective students' post-experience group mean was significantly higher than the control dental students' mean (P=0.02, ANOVA) which reflects significant (P<0.05) increases in 9 of 13 AEGD elective students' individual survey means compared to 4 of 18 control dental students' means. Both groups had pre-experience means of 2.9 or below on the same 9 statements, indicating poor self-rated skills. Four of 9 AEGD elective students' post-experience means significantly (P<0.05) increased, while only 1 of 9 control dental students' means increased. The data suggests that AEGD elective students were more confident in their clinical abilities and patient management skills than control dental students and they also reported greater improvement in low-rated skills. When adequate parameters were maintained that enhanced the accuracy of self-assessment, students appeared to rate their skill levels seriously. Independent self-assessment may be an important component of life-long learning. PMID- 11872070 TI - Psychological stress in undergraduate dental students: baseline results from seven European dental schools. AB - OBJECTIVES: To determine the degree of psychological distress, the experience of emotional exhaustion, and the extent of stress associated with course work in dental students and to compare these measurements among seven European dental schools. DESIGN: Multi-centred survey. SETTING: Dental Schools at Amsterdam, Belfast, Cork, Greifswald, Helsinki, Liverpool and Manchester. PARTICIPANTS: 333 undergraduate first-year dental students. MEASURES: General Health Questionnaire (GHQ12), Maslach Burnout Inventory (MBI), Dental Environment Stress Questionnaire (DES), demographic variables. PROCEDURE: Questionnaire administered to all students attending first year course. Completed questionnaires sent to central office for processing. RESULTS: Seventy-nine percent of the sampled students responded. Over a third of the students (36%) reported significant psychological distress (morbidity) at the recommended cut-off point (>3 on GHQ). These scores were similar to those reported for medical undergraduates. Twenty-two percent recorded comparatively high scores on emotional exhaustion. A wide variation in these 2 measurements was found across schools (p's<0.001). Stress levels indicated by the DES were less variable (p>0.5). Some evidence showed that contact with patients and the level of support afforded by living at home may be protective. CONCLUSION: Higher than expected levels of emotional exhaustion were found in a large sample of first-year undergraduate dental students in Europe. PMID- 11872071 TI - Computer literacy and attitudes among students in 16 European dental schools: current aspects, regional differences and future trends. AB - A questionnaire survey was carried out to investigate the competence and attitude of dental students towards computers. The current study presents the findings deriving from 590 questionnaires collected from 16 European dental schools from 9 countries between October 1998 and October 1999. The results suggest that 60% of students use computers for their education, while 72% have access to the Internet. The overall figures, however, disguise major differences between the various universities. Students in Northern and Western Europe seem to rely mostly on university facilities to access the Internet. The same however, is not true for students in Greece and Spain, who appear to depend on home computers. Less than half the students have been exposed to some form of computer literacy education in their universities, with the great majority acquiring their competence in other ways. The Information and Communication Technology (ICT) skills of the average dental student, within this limited sample of dental schools, do not facilitate full use of new media available. In addition, if the observed regional differences are valid, there may be an educational and political problem that could intensify inequalities among professionals in the future. To minimize this potential problem, closer cooperation between academic institutions, with sharing of resources and expertise, is recommended. PMID- 11872073 TI - Evaluation of an undergraduate community-based course in Family Dentistry. AB - This study aimed to determine dental students' expectations of attending a community clinic to provide family dental care and to assess how well the course met their expectations. Prior to the start of the course, 30 final year students were asked about the type and number of patients they expected to treat, the types of treatment they thought they would provide and additional skills, if any, they might gain and also about any worries they had about the course. The responses were used to compile a questionnaire, completed by the students after finishing the course. The students treated more patients a day than expected and underestimated the range of treatments carried out at the clinics, particularly trauma and advanced restorative work. Additional communication skills and increased confidence and quality of work were among the cited gains. Concerns expressed prior to the course had largely been eliminated, although some students still had reservations about National Health Service paperwork, treatment planning and running late. It was concluded that The Family Dentistry Course more than fulfilled students' expectations and it gave them enhanced skills in patient management and clinical competency. PMID- 11872074 TI - Special report: ADEE INITIATIVE. The positive tendency in approaches to the dental curriculum in Belarus: a report arising from a WHO-sponsored visitation to schools in the former USSR and Eastern Europe and a follow-up invited visit of ADEE/AADS representatives to Minsk, Belarus. PMID- 11872072 TI - Continuing professional development amongst dental practitioners in the United Kingdom: how far are we from lifelong learning targets? AB - This study was conducted to identify the extent to which qualified dental practitioners in the UK currently undertake three distinct activities of Continuing Professional Development (CPD): reading professional journals; attending courses; and undertaking retraining courses. Also, to determine the impact of gender, age, length of time since qualification, current working hours and career breaks upon the extent to which dental practitioners engage in CPD. Data were analysed from a questionnaire survey of a systematic sample of one in 10 dentists taken from the Dentists Register of the UK General Dental Council. The response rate was 66.6%. Only dentists practising at the time of the survey were included in the analysis (N = 1550). A high proportion of the sample reported regularly undertaking activities related to CPD. Approximately 87% read professional journals at least once per month; just over half had attended five or more days at professional meetings and courses in the last year. Only a small proportion of dentists (12%) had undertaken a retraining course in the past three years. Those dentists who had been qualified for between 21 and 30 years, those who had gained additional qualifications after qualifying as a dentist, and those who had taken a career break at some point in their life were more likely to read professional journals. Attendance at postgraduate dental courses was related to being male, not having taken a career break, possessing an additional qualification, longer working hours, and not being a General Dental Practitioners (GDP). Attendance at a retraining course was less likely for those who had taken a career break, those who had attended fewer courses in the last year and for those who worked 30--40 h per week. A large proportion of dental practitioners are currently undertaking sufficient CPD to meet the UK requirements of recertification. Greater attention should be directed towards identifying the barriers to CPD among female dentists and those who have been qualified for longer periods. More research is needed in assessing the long-term CPD consequences of those who have returned to dentistry following a career break. PMID- 11872075 TI - Prognostic factors in advanced stage Hodgkin's lymphoma: the significance of the number of involved anatomic sites. AB - BACKGROUND: Advanced Hodgkin's lymphoma (HL) is curable by conventional chemotherapy in 60--70% of patients. The pretreatment identification of a sizeable subgroup of patients with sufficiently low failure-free survival (FFS) to be eligible for investigational treatment is necessary. OBJECTIVES: To determine the prognostic significance of the number of involved sites (NIS) in patients with advanced HL and its relationship to the International Prognostic Score (IPS). METHODS: A retrospective review of patients with advanced HL, defined as Ann Arbor stage (AAS) IB, IIB, III or IV, treated with anthracycline based regimens. The end-point was FFS. RESULTS: We identified 277 patients with a median age of 32 yr (14--78), 57% of whom were males. AAS was I in 4% of patients, II in 29%, III in 38% and IV in 29%. B-symptoms were recorded in 81%. Most patients had nodular sclerosis (64%) and mixed cellularity (26%) histology. IPS was greater-than-or-equals 3 in 44% of 242 evaluable patients. The NIS was greater-than-or-equals 5 in 32% of the patients and 20% of all patients had both greater-than-or-equals 5 involved sites and IPS greater-than-or-equals 3. The 10 yr FFS was 67%, being 76% vs. 50% for patients with less-than-or-equals 4 vs. greater-than-or-equals 5 involved sites (P < 0.0001). The NIS (greater-than-or equal 5), AAS IV and anemia were independent predictors of FFS in multivariate analysis. The NIS remained significant along with IPS, when the latter was included in the analysis. Patients with greater-than-or-equals 5 involved sites and IPS greater-than-or-equals 3 had 10-yr FFS overall, and relapse-free survival of 41%, 45% and 49%, respectively. CONCLUSIONS: The NIS was associated with FFS in advanced HL, was independent of IPS, and led to the identification of a sizeable subgroup of patients with 10-yr FFS of approximately 40%. This factor should be evaluated during the development of prognostic systems. PMID- 11872076 TI - Mobilisation of tumour cells along with CD34+ cells to peripheral blood in multiple myeloma. AB - BACKGROUND: Cells belonging to the malignant clone are found in the peripheral blood in myeloma patients. In order to minimise the content of tumour cells in the stem cell product it is crucial to perform stem cell harvest at a time when tumour cells in the peripheral blood are at a minimum. OBJECTIVE: The aim of the study was to compare the mobilisation kinetics of normal CD34+ cells and myeloma plasma cells during mobilisation with either G-CSF alone or high-dose cyclophosphamide (HDCy) plus G-CSF. DESIGN AND METHODS: Morning blood samples were drawn each day during mobilisation from start of G-CSF or HDCy and to the end of leukapheresis, and were analysed by flow cytometry for content of CD34+ cells and myeloma plasma cells (CD38+ + CD45-). Tumour cells were also estimated by a patient-specific real-time polymerase chain reaction (PCR) method based on the 5' nuclease TaqMan technology. RESULTS: Flow cytometry data from 16 patients showed concomitant mobilisation of CD34+ cells and myeloma plasma cells. Seven patients were mobilised twice; first with G-CSF alone and then with HDCy plus G CSF. There was no difference between the two mobilisation regimens regarding tumour cell mobilisation kinetics. Real-time PCR was performed in one patient and confirmed the mobilisation of tumour cells at the time when CD34+ blood cells were at a maximum. CONCLUSIONS: Tumour cells are mobilised to the peripheral blood at the same time as CD34+ cells in multiple myeloma patients after priming with both G-CSF alone and HDCy in combination with G-CSF. PMID- 11872077 TI - Frequency and prognostic relevance of cyclin D1 dysregulation in multiple myeloma. AB - OBJECTIVE: Cyclin D1 dysregulation has been found with varying frequencies in multiple myeloma (MM) and has been suggested to be associated with a poor prognosis. The aim of this study was to investigate the frequency of cyclin D1 dysregulation in patients being treated for MM and to test whether cyclin D1 dysregulation is a prognostic factor for MM patients. METHODS: To achieve the above aims we designed a highly sensitive and reproducible real-time reverse transcription polymerase chain reaction (RT-PCR) assay for quantitation of cyclin D1 mRNA. Using this assay, 110 diagnostic bone marrow (BM) samples from patients with MM were screened for cyclin D1 dysfunction. RESULTS: The real-time assay was able to detect the presence of 0.01% cyclin D1 positive cells allowing a safe detection in MM BM samples. In 42% (46/110) of MM BM samples a greater-than-or equals 3-fold increase in cyclin D1 mRNA was observed compared to the cyclin D1 level in normal BM. In the remaining group of MM patients the cyclin D1 mRNA levels were comparable to normal donors. Follow-up of 76 MM patients showed no significant (P = 0.35) difference in survival between cyclin D1 positive and negative MM patients. In addition, cyclin D1 dysregulation did not correlate with known prognostic factors. CONCLUSION: The developed real-time RT-PCR assay for detection of cyclin D1 mRNA levels offers a fast and safe screening for cyclin D1 dysfunction. When a large cohort of MM patients was screened, the cyclin D1 gene was found to be frequently dysregulated, but there was no significant correlation to survival or known prognostic parameters. PMID- 11872078 TI - Kinetics of BCR-ABL fusion transcript levels in chronic myeloid leukemia patients treated with STI571 measured by quantitative real-time polymerase chain reaction. AB - The activated tyrosine kinase, which arises as a result of the balanced t(9,22) translocation in chronic myeloid leukemia (CML), is thought to be essential for the development of the leukemic phenotype. Recently, designer drugs have been introduced which specifically inhibit such specific kinases. Among these, STI571 (Glivec) has entered clinical trials and shown promising activities in chronic phase (CP), accelerated phase (AP) and blast crisis (BC) as evidenced by significant hematological and cytogenetic responses in CML patients. To evaluate the effect of STI571 at the molecular level we have employed quantitative real time PCR (RQ-PCR) to measure the amount of BCR-ABL fusion transcript in a series of 19 patients treated with STI571, either in CP(11) or in (AP)(8) of the disease for 3--9 months (median 6 months). Employing this method, which is able to detect at least one BCR-ABL+ cell in 500,000, in serial blood and bone marrow specimens we found decreases in transcript levels in 10/11 CP patients, but only in 1/8 of the AP patients. When present such decreases were gradual and became evident only after 3 months of STI571 treatment, and their kinetics in blood closely mirrored those seen in parallel marrow samples. Moreover, decreases were between 10- and 100-fold in 11/13 patients, with only two patients reaching residual disease levels below 10(-2) (a 900-fold decrease). Thus, no patient reached PCR negativity. We conclude that the RQ-PCR method is a highly suitable tool for following the effect of STI571 in CML and that further validation of the method, performed in a prospective manner, will contribute significantly to the elucidation of the proper role of STI571 in CML. PMID- 11872079 TI - Telomerase regulation and telomere dynamics in germinal centers. AB - Telomere length maintenance, usually executed by telomerase, is a prerequisite for an extended or infinite division potential. Nevertheless most telomerase positive normal cells exhibit telomere shortening. This study details the telomerase expression and telomere dynamics in purified tonsil B cell subsets during the germinal center (GC) reaction. Significant telomere lengthening was observed as naive B cells matured to centroblasts and when centroblasts matured further to centrocytes, resulting in an increase in telomere length of about 4 kbp determined by Southern blotting. Immunopurified cell populations were also studied by fluorescence in situ hybridization and flow cytometry (flow-FISH) confirming that the GC B cells exhibited lengthened telomeres. These data were further verified in unpurified tonsil cells by combining flow-FISH and immunophenotyping using selected surface markers. Centroblasts expressed high levels of telomerase activity, which was increased in centrocytes, whereas resting naive, activated naive and memory B cells were telomerase activity negative. Expression levels of the catalytic subunit (hTERT) RNA paralleled the telomerase activity levels. The unique telomere elongation in GC B cells permits extensive proliferation during the GC reaction and provides the memory cells with a substantial increase in division potential. Understanding the telomere biology of GC cells is important in defining requirements for telomere elongation in vivo, with implications for the normal immune system as well as for lymphomas, and could provide insights into how the division potential of cells can be manipulated in vitro. PMID- 11872080 TI - Bronchiolitis obliterans organizing pneumonia in a patient with chronic myelogenous leukemia developing after initiation of interferon and cytosine arabinoside. AB - A 59-yr-old man developed fevers, shortness of breath, persistent cough and weight loss, shortly after initiation of therapy with interferon-alpha 2a and cytosine arabinoside for treatment of chronic myelogenous leukemia. Radiologic pulmonary infiltrates and lung tissue biopsy were consistent with bronchiolitis obliterans organizing pneumonia (BOOP). After discontinuation of the chemotherapeutic drugs, the pneumonic symptoms and chest roentgenogram infiltrates resolved. This report suggests that treatment with interferon-alpha, in combination with cytosine arabinoside, may produce the rare complication of BOOP. PMID- 11872081 TI - Plasmablastic lymphoma in a patient with chronic lymphocytic leukemia heavily pretreated with cladribine (2-CdA): an unusual variant of Richter's syndrome. AB - Patients with chronic lymphocytic leukemia (CLL) may develop a large-cell transformation known as Richter's syndrome (RS). RS usually presents as diffuse large-cell lymphoma (DLCL) or its immunoblastic variant, and it can be recognized simultaneously with CLL or even 23 yr after its diagnosis. We describe an unusual case of CLL treated with cladribine (2-CdA) in whom DLCL of the plasmablastic type (PBL) developed 4 yr after CLL (Rai IV) diagnosis and 1.5 yr after the 10th course of 2-CdA treatment. Immmunologic, cytogenetic, and molecular studies performed at the time of CLL and PBL coappearance indicated that both tumors originated from different B-cell progenitors. Both malignancies were refractory to VAD (vincristine, doxorubicin, dexamethasone)-based chemotherapy, and only partial response was achieved with CHOP (cyclophosphamide, doxorubicin, vincristine, prednisone) salvage treatment. However, the patient died 6 months after the occurrence of RS due to rapid progression of PBL. This is the first description of a CLL patient who developed an unusual plasmablastic variant of RS. Recently, the PBL entity has been identified among DLCL associated with the human immunodeficiency virus (HIV) infection. We suggest that in our CLL patient heavily pretreated with 2-CdA, PBL arose as a second clone due to the prolonged and severe state of the host's immunosuppression. Overall survival with current strategies is poor, and further insight into the natural history, biology, and treatment of PBL are needed. PMID- 11872082 TI - Skin lesion caused by hydroxyurea. PMID- 11872083 TI - Hereditary giant platelet disorder presented as pseudothrombocytopenia. PMID- 11872084 TI - MAGE-C1 (CT7) gene expression in multiple myeloma: relationship to sperm protein 17. PMID- 11872085 TI - 'The Mexican approach' to conduct NST. PMID- 11872086 TI - Chemokines--linking receptors to response. PMID- 11872089 TI - Chemokines in allergic lung inflammation. PMID- 11872087 TI - Chemokine signalling: pivoting around multiple phosphoinositide 3-kinases. AB - The role of chemokines in mediating directional cell migration is well established, but more recently it has become evident that chemokines are able to couple to distinct signalling pathways that are involved in not only chemotaxis, but also cell growth and transcriptional activation. The signalling pathway controlled by the phosphoinositide 3-kinase (PI3K) family of lipid kinases has been the focus of much attention with respect to their role in chemokine-mediated functional responses. Indeed, there now exists convincing biochemical, pharmacological and genetic evidence that both CC and CXC chemokines stimulate PI3K-dependent chemotaxis of inflammatory cells such as eosinophils, macrophages, neutrophils and T lymphocytes. This review considers the role of individual PI3Ks (e.g. the p85/p110 heterodimer, PI3Kgamma and PI3KC2alpha) as well their downstream effector targets in mediating chemokine-stimulated cell migration. PMID- 11872088 TI - Role of chemokines and chemokine receptors in the gastrointestinal tract. AB - The pathological association between leucocytes and gastrointestinal diseases has long been recognized. Chemokines are a large family of chemotactic cytokines whose fundamental role is the recruitment of leucocytes to tissues. Although chemokines and their receptors are considered to be mediators of inflammation and tissue injury in several inflammatory diseases, their precise role in the pathophysiology of gastrointestinal diseases remains incompletely understood. Nonetheless, by virtue of their expression and localization at sites of gastrointestinal tissue injury and inflammation, a number of investigators have suggested a vital role for chemokines and their receptors in the pathophysiology of gastrointestinal diseases. This short review examines the role of chemokines and their receptors in the gastrointestinal tract with an emphasis on their involvement in the regulation of intestinal and hepatic inflammation. PMID- 11872090 TI - CXCR4 expression on monocytes is up-regulated by dexamethasone and is modulated by autologous CD3+ T cells. AB - Chemokines and their receptors regulate cell migration to sites of inflammation. The glucocorticoid dexamethasone has potent anti-inflammatory effects, yet paradoxically up-regulates expression of some cytokine receptors. We have examined the effects of dexamethasone on chemokine receptor expression. Using an RNase protection assay, we show that dexamethasone up-regulates human peripheral blood mononuclear cell (PBMC) expression of CXCR4 mRNA. Flow cytometric analysis demonstrated that increased expression of CXCR4, but not CXCR1 and CXCR2, occurred on both monocytes and CD3+ T cells in PBMC mixed cultures. A stromal derived factor (SDF)-1alpha-mediated calcium influx was detected on monocytes. Basal levels of CXCR4 expression on purified monocytes were lower when compared with monocytes in mixed PBMC cultures. Co-culture of monocytes with purified CD3+ T cells led to enhanced basal expression of CXCR4 on monocytes. The use of transwells to partition CD3+ T cells resulted in increased CXCR4 expression on monocytes, suggesting that CD3+ T-cell derived soluble factors regulate CXCR4 expression. PMID- 11872091 TI - Induction of thymocyte positive selection does not convey immediate resistance to negative selection. AB - The acquisition of functional competence represents a critical phase during intrathymic development of T cells. Thymocytes reaching this stage represent cells which have been positively selected on the basis of major histocompatibility complex reactivity, but which have also been purged of potentially autoreactive T-cell receptor specificities by negative selection. While the developmental window in which thymocytes are subjected to positive selection is now well defined, the precise developmental timing of negative selection, in relation to positive selection events, is less clear. Moreover, the underlying mechanism allowing single-positive thymocytes to respond to T-cell receptor ligation by activation rather than death, remains controversial. Here we have analysed the developmental timing of negative selection in relation to positive selection, using measurement of thymocyte susceptibility to dendritic cell presentation of the superantigen staphylococcal enterotoxin B (SEB). We show that thymocytes which have received initial positive selection signals, namely CD4+ CD8+ CD69+ thymocytes, like their CD4+ CD8+ CD69minus sign precursors, are susceptible to negative selection, indicating that induction of positive selection does not convey immediate resistance to negative selection. In contrast, newly generated CD4+ CD8minus sign CD69+ cells are not only resistant to deletion by SEB, but respond to SEB-mediated T-cell receptor-ligation by activation, indicating that the acquisition of functional competence occurs at the newly generated CD4+ CD8minus sign CD69+ stage. Finally, by using direct retroviral infection of primary CD4+ CD8+ thymocytes, we also show that Notch-1 activation in CD4+ CD8+ thymocytes does not correlate with, nor convey resistance to superantigen-mediated negative selection. Thus, our data suggest that although Notch-1 has been implicated in resistance to thymocyte apoptosis, the acquisition of resistance to negative selection occurs independently of Notch-1 signalling. PMID- 11872092 TI - Roles of cytotoxic T-lymphocyte-associated antigen-4 in the inductive phase of oral tolerance. AB - To elucidate the roles of cytotoxic T-lymphocyte-associated antigen-4 (CTLA-4) in oral tolerance, we studied the consequences of CTLA-4 blockade during the inductive phase of oral tolerance using a transgenic T-cell transfer model. We found that CTLA-4 blockade significantly accelerated cell cycle progression of antigen-specific T cells and dramatically increased their numbers in lymphoid organs following oral administration of ovalbumin (OVA). In mice fed with OVA, only approximately 35% of specific T cells underwent more than four cycles of cell division. This was increased to 65% in mice fed with OVA and treated with a blocking anti-CTLA-4 monoclonal antibody (mAb). The OVA-specific T cells in the latter group were localized primarily in the T-cell zones of the mesenteric lymph nodes and Peyer's patches with a few penetrated into B-cell follicles. Nevertheless, both faecal anti-OVA immunoglobulin A (IgA) and seral anti-OVA immunoglobulin G (IgG) were produced in anti-CTLA-4 mAb-treated mice. These results suggest that CTLA-4 limits the degree of T-cell activation by blocking cell cycle progression during the inductive phase of oral tolerance. In the absence of the CTLA-4 signal, mucosal exposure of antigen induces heightened T cell activation and expansion, which in turn promotes the production of antigen specific antibodies. PMID- 11872093 TI - Immunological characterization of a gammadelta T-cell stimulatory ligand on autologous monocytes. AB - Bovine gammadelta T cells are stimulated to proliferate by autologous monocytes. This is referred to as the autologous mixed leucocyte reaction (AMLR). It has been shown previously that the stimulatory component is constitutively expressed on the monocyte plasma membrane and is a protein or has a protein moiety. Here we showed that gammadelta T-cell responses to the monocytes requires interaction with the T-cell receptor because Fab1 fragments of a monoclonal antibody (mAb) that reacts with the delta chain of the T-cell receptor blocked proliferation in the AMLR. Monocyte molecules involved in stimulation were also characterized further by biochemical and immunological methods. A mAb, named M5, was generated by immunizing mice with bovine monocytes and shown to block the ability of monocytes to stimulate in the AMLR. Treatment of monocytes or monocyte membranes with high salt, chelating agents or phospholipase C did not affect their ability to stimulate gammadelta T-cell proliferation or reactivity with mAb M5 indicating the ability of monocytes to stimulate does not involve peripheral membrane components or a glycosyl-phosphatidylinsositol (GPI)-anchored components. Hence it was concluded that the stimulation occurred as a result of intergral membrane proteins including that recognized by mAb M5. The ligand for mAb M5 was on all bovine monocytes and to a lower level on granulocytes but not on lymphocytes. MAb M5 also reacted with sheep monocytes but not with human monocytes or murine macrophages, in agreement with a previous reports that sheep monocytes but not human or mouse mononuclear phagocytes have the capacity to stimulate bovine gammadelta T cells in in vitro cultures. The level of expression of the M5 ligand was not altered by gamma-irradiation or culture of monocytes with lipopolysaccharide but it was decreased following culture with interferon-gamma containing cell culture supernatants. PMID- 11872094 TI - Expression of caveolin by bovine lymphocytes and antigen-presenting cells. AB - Caveolin is a generic term for a family of proteins that include caveolin-1, -2 and -3. Although the distribution of these proteins varies between cells, caveolin-1 and -2 are commonly found coating membrane invaginations known as caveolae. Studies on human and murine cells suggest that caveolin/caveolae can be found in neutrophils, macrophages and mast cells, in which they are involved in the uptake of pathogens, but not in lymphocyte cell lines. Expression of caveolin 1, -2 and -3 in bovine immune cells was investigated using confocal microscopy and Western blotting. Staining for caveolin-1 was evident in all peripheral blood mononuclear cells (PBMC) and in CD4+, CD8+ and CD21+ lymphocytes, monocytes, macrophages and monocyte-derived dendritic cells (DC). In addition, the caveolin 1 antibody detected a protein with a molecular weight of approximately 22[?]000 in all PBMC, macrophages and DC, as well as in bovine aortic endothelial (BAE)-1 cells and human endothelial cells by Western blotting. In macrophages and DC, caveolin co-localized with the endoplasmic reticulum--Golgi intermediate compartment (ERGIC) and to a lesser extent with Golgi, but not with endoplasmic reticulum. Staining was not seen on the plasma membrane in any bovine immune cells, suggesting the absence of caveolae, while in BAE-1 cells staining was predominantly on the cell membrane. Caveolin-2 could not be detected in any bovine cells by confocal microscopy or Western blotting, while caveolin-3 was detected in all bovine cells by Western blotting, but not by confocal microscopy. These data provide evidence for the presence of caveolin in bovine lymphocytes and antigen-presenting cells. PMID- 11872095 TI - Age-dependent increase of peritoneal B-1b B cells in SCID mice. AB - The impact of increasing age upon immunoglobulin production and B-lymphocyte generation in "leaky" severe combined immune-defective (SCID) mice was examined by enzyme-linked immunosorbent assay and flow cytometry. By 1 year of age, the mice had normal numbers of B cells in their peritoneal cavity, while their spleen had very few immunoglobulin M-positive (IgM+) cells. The majority of B cells expressed the CD11b marker characteristic of the B-1b subset. B-1a (CD5+) cells were present at a lower frequency and B-2 cells were absent. The frequency of mice producing detectable immunoglobulin increased with age, and isotype diversity within individual mice was variable. IgM production was most frequently observed followed by IgG3 and IgG2a, then IgG1, and finally IgA. The selective persistence of the B-1 B-cell subset in the peritoneal cavity of aging SCID mice is a natural model for the study of those genetic and environmental influences that determine lymphocyte longevity. PMID- 11872096 TI - Immunization onto bare skin with synthetic peptides: immunomodulation with a CpG containing oligodeoxynucleotide and effective priming of influenza virus-specific CD4+ T cells. AB - Exploiting the immune system of the skin for vaccine administration offers an attractive alternative to the currently used invasive immunization procedures. In this study we report that a synthetic peptide representing a T-helper (Th) epitope from influenza virus haemagglutinin (aa 307--319) can be an effective immunogen when coapplied with cholera toxin (CT) onto bare skin. Proliferation of both peptide- and influenza virus-specific CD4+ T cells was measured in lymphocyte cultures from spleens and regional lymph nodes. The presence of the CpG oligodeoxynucleotide 1826 in the peptide/CT formulation, enhanced the proliferation of peptide- and virus-specific T cells as measured by the conventional [(3)H]thymidine uptake and interleukin (IL)-2 assays. Furthermore, the bias towards Th2-type of responses stimulated by CT was shifted towards Th1 as demonstrated (i) by the increase of interferon-gamma and decrease of IL-4 cytokine levels measured in culture supernatants, (ii) by the predominance of IG2a anti-CT antibodies in the serum, and (iii) by the down-regulation of total serum IgE antibody levels. These findings demonstrate the potential of the bare skin as a non-invasive route for administration of small molecular size peptide antigens. Furthermore, with the selection and combination of the appropriate type of adjuvants, immune responses can be modulated towards the desired type of Th phenotype. PMID- 11872097 TI - Fc receptor regulation of protective immunity against Chlamydia trachomatis. AB - The prevailing paradigm for designing potentially efficacious vaccines against the obligate intracellular bacterium, Chlamydia trachomatis, advocates regimens capable of inducing a mucosal antigen-specific T helper type 1 (Th1) response. However, recent reports indicate that rapid and efficient clearance of a secondary infection also requires certain B-cell functions. We investigated the hypothesis that Fc receptor (FcR)-mediated antibody effector mechanisms are important B-cell-related functions involved in controlling a chlamydial genital reinfection. Microbiological analysis of genital chlamydial infection in FcR knockout (FcRKO) mice lacking the activatory FcgammaRI (CD64) and FcRgammaIII (CD16), as well as the inhibitory FcgammaRIIB1 (CD32), revealed a greater intensity of secondary infection (i.e. bacterial shedding) in FcRminus sign/minus sign as compared to FcR+/+ mice; however, the course of the primary infection was indistinguishable in both animals. Pathologically, FcRKO mice suffered greater ascending infection than immunocompetent wild-type (WT) mice after a secondary infection. Immunological evaluation indicated that the presence of specific anti chlamydial antibodies enhanced chlamydial antigen presentation for induction of a Th1 response by FcR+/+, but not FcRminus sign/minus sign, antigen-presenting cells. In addition, specific anti-chlamydial antibodies augmented both macrophage killing of infected epithelial cells by antibody-dependent cellular cytotoxicity (ADCC) and macrophage inhibition of productive growth of chlamydiae in co cultures. These results indicate that B cells participate in anti-chlamydial immunity via FcR-mediated effector functions of antibodies, which are operative during reinfections. Such effector functions include ADCC, and possibly enhanced uptake, processing and presentation of chlamydial antigens for rapid induction of a Th1 response, all facilitating the early clearance of an infection. These findings suggest that a future anti-chlamydial vaccine should elicit both humoral and T-cell-mediated immune responses for optimal memory response and vaccine efficacy. PMID- 11872098 TI - Mycobacterial antigens induce apoptosis in human purified protein derivative specific alphabeta T lymphocytes in a concentration-dependent manner. AB - The morbidity and lethality of tuberculosis is partially the result of an ineffective delayed-type hypersensitivity reaction which causes caseating granulomas in the lung and other organs. Recently we showed that during caseation besides macrophages numerous Fas+ FasL+ lymphocytes undergo apoptosis and postulated that this phenomenon may be due to activation-induced cell death (AICD) as a consequence of T-lymphocyte reactivation via bacillary antigens. As purified protein derivative of Mycobacterium tuberculosis (Mtb-PPD) provokes caseation in tuberculosis patients, the question arose as to whether bacillary antigens are responsible for AICD within caseous areas. In the present study Mtb PPD-specific T helper 1 (Th1)-differentiated T lymphocytes were generated in vitro. Reactivation of these cells with Mtb-PPD resulted in a concentration dependent hyporesponsiveness, which was due to an increase in apoptosis of gammadelta+, alphabeta+ CD4+ as well as alphabeta+ CD8+ T lymphocytes as assessed by the demonstration of the apoptosis-associated mitochondrial membrane protein 7A6 and DNA fragmentation. Blocking experiments demonstrated that Mtb-PPD antigens exploited the Fas/FasL system to induce apoptosis in Mtb-PPD-specific T lymphocytes. These results may support the hypothesis that in tubercle granulomas with caseation T lymphocytes undergo AICD following reactivation by bacillary antigens, thus contributing to the persistence of tuberculosis. PMID- 11872101 TI - Intuition and the development of expertise in surgical ward and intensive care nurses. AB - AIM: The aim of this study was to explore and identify nurses' clinical expertise in surgical ward and intensive care settings in England. One of the objectives of the study and the focus of this paper was the exploration of these nurses' understanding and use of intuition in the context of their practice. BACKGROUND: Since 1980 many studies have investigated qualified nurses' use of intuition within aspects of their practice. However, it was Benner's (1982, 1984) seminal work that firmly established the relationship between intuition and expert clinical practice. Since that time a possible relationship between intuitive components of decision-making and nonexpert nurses' practice has remained relatively unexplored until recently. METHODS: This constructivist qualitative study incorporated observation and interview to explore 61 qualified nurses' expertise through their assessment of patients following major surgery. FINDINGS: Specific findings highlighted refinement in nurses' use of intuitive and analytical elements of decision-making across the four identified levels of expertise. The most fluent and effective use of intuitive and analytical components of decision-making was found in the expert group. CONCLUDING REMARKS: These results are discussed in relation to current understanding of the components of expert decision-making in nursing practice. Both intuitive and analytical elements should be recognized in any model that seeks to depict the true nature of nurses' decision-making as they develop clinical expertise. PMID- 11872099 TI - Interleukin-10 promotes B16-melanoma growth by inhibition of macrophage functions and induction of tumour and vascular cell proliferation. AB - The aim of this study was to investigate the mechanisms by which interleukin-10 (IL-10) induces tumour growth in a mouse-melanoma model. A B16-melanoma cell line (B16-0) was transfected with IL-10 cDNA and three clones that secreted high (B16 10), medium and low amounts of IL-10 were selected. Cell proliferation and IL-10 production were compared in vitro, and tumour growth, percentages of necrotic areas, tumour cells positive for proliferating cell nuclear antigen (PCNA), IL-10 receptor (IL-10R) and major histocompatibility complex type I (MHC-I) and II (MHC II), as well as infiltration of macrophages, CD4+ and CD8+ lymphocytes and blood vessels were compared in vivo among IL-10-transfected and non-transfected tumours. Proliferation and tumour growth were greater for IL-10-transfected than for non-transfected cells (P < 0.001), and correlated with IL-10 concentration (r > or =0.79, P < 0.006). Percentages of tumour cells positive for PCNA and IL-10R were 4.4- and 16.7-fold higher, respectively, in B16-10 than in B16-0 tumours (P < 0.001). Macrophage distribution changed from a diffuse pattern in non transfected (6.4 +/- 1.7%) to a peripheral pattern in IL-10-transfected (3.8 +/- 1.7%) tumours. The percentage of CD4+ lymphocytes was 7.6 times higher in B16-10 than in B16-0 tumours (P = 0.002). The expression of MHC-I molecules was present in all B16-0 tumour cells and completely negative in B16-10 tumour cells. In B16 0 tumours, 89 +/- 4% of the whole tumour area was necrotic, whereas tumours produced by B16-10 cells showed only 4.3 +/- 6% of necrotic areas. IL-10 transfected tumours had 17-fold more blood vessels than non-transfected tumours (61.8 +/- 8% versus 3.5 +/- 1.7% blood vessels/tumour; P < 0.001). All the effects induced by IL-10 were prevented in mice treated with a neutralizing anti IL-10 monoclonal antibody. These data indicate that IL-10 could induce tumour growth in this B16-melanoma model by stimulation of tumour-cell proliferation, angiogenesis and immunosuppression. PMID- 11872102 TI - Intravascular administration sets are accurate and in appropriate condition after 7 days of continuous use: an in vitro study. AB - BACKGROUND: The ideal duration of intravascular administration set use is unknown. Studies have compared the infective implications of 1--7 days of use. The Centers for Disease Control recommend at least 3 days usage. No previous study has evaluated the accuracy of volume delivery or integrity of administration sets after prolonged use. AIM: To evaluate the accuracy and condition of intravascular administration sets used continuously for 7 days. DESIGN: Prospective, randomized, experimental study in the laboratory setting. METHODS: Four administration sets were randomly assigned to deliver 2 mL/hour (IMEDreg syringe set 2280--0000), 20, 50 or 100 mL/hour (IMEDreg infusion sets 2210--0500) of crystalloid solution continuously for 7 days through an IMEDreg Geminireg four channel infusion pump (PC4). At study commencement and daily for 7 days, a 4-hour volume measurement and an inspection for leaks/erosion of administration sets occurred for each administration set (total measurements = 32). RESULTS: Mean volume outputs over 4 hours were 7.84 mL (2 mL/hour), 80.66 mL (20 mL/hour), 205.35 (50 mL/hour) and 406.37 (100 mL/hour). These differed significantly from the programmed volumes (P=0.00--0.01). Usage duration did not influence performance (F=0.866, P=0.55). Accuracy of volume delivery differed significantly with pump speed (F=106.933, P < 0.001) exhibiting increased volume to 50 mL/hour then a reduction at 100 mL/hour. Differences were within manufacturer specifications (+/-5%) and were clinically acceptable. All administration sets remained in appropriate condition displaying no leakage or erosion. CONCLUSION: There were small inaccuracies found between programmed and delivered volumes, however, there was no deterioration in performance over time. This suggests that inaccuracies were because of normal pump performance rather than the administration sets. Administration sets retain acceptable accuracy and condition after 7 days continuous use. Further research should assess the infective and other impacts of prolonged usage. PMID- 11872103 TI - The meaning of mobility for residents and staff in long-term care facilities. AB - BACKGROUND: Although the physical and psychological effects of decreased mobility have been documented, little is known about what it means to be mobile from the institutionalized elders' perspective. Even less is known about nurses' perceptions regarding institutionalized elders' mobility and about mobility enhancing strategies. AIM: The purpose of this study was to generate knowledge about the phenomenon of mobility from both the nurses' and institutionalized elders' perspective. DESIGN: An exploratory qualitative design was used. Focus groups with residents and nursing staff were conducted in three long-term care facilities in a large metropolitan city in 1998. Ethical approval was received from the ethics committees in all three facilities prior to study implementation. Twenty long-term care facility residents and 15 nursing staff participated in the study. METHODS: A trained facilitator used an interview guide to moderate discussions in which residents' and nurses' opinions and feelings about mobility were solicited. The group responses were tape recorded, transcribed and coded. The codes reflected major concepts or abstractions of the data and emerged from the participants' verbal responses to the research questions. FINDINGS: Both groups identified mobility as being pivotal to the residents' quality of life and well-being. The residents viewed mobility as a means of freedom, choice and independence, and they made great efforts to maintain their mobility. The nurses also valued the importance of mobility and assisted the residents through preparatory care, environmental modifications and encouragement. Factors that related to the residents' willingness to be mobile, environmental barriers and the impact of waiting were compared between the two groups. CONCLUSION: Mobility involves more than the movement from one point to another. The nature of the assistive nurse-client relationship and the resident's subjective perspective must become central to understanding the meaning of mobility for residents in a long-term care facility. PMID- 11872105 TI - An insight into the experiences of parents with inflammatory bowel disease. AB - AIMS AND OBJECTIVES: To identify parents' views, both positive and negative on: how inflammatory bowel disease (IBD) affects people in their parenting role; effects parents with IBD have noticed in their children; ways of dealing with any difficulties in parenting; and support needed by parents with IBD. DESIGN: This was a small-scale exploratory study, adopting a qualitative research design. A purposive sample of 19 mothers and five fathers with IBD was recruited through two hospitals and a voluntary group in the north of England. RESEARCH METHODS: Data were collected through a combination of focus groups and interviews, and analysed using the 'framework approach'. FINDINGS: The main positive effect for parents was developing a closer relationship with their children. When parents were experiencing symptoms there were difficulties in caring for young children; problems taking children to and from school, and attending school events; restrictions in social life; irritability and reduced tolerance of children. When in hospital, it was sometimes difficult to arrange for care of young children. These difficulties caused worry and guilt for parents, and a few spoke of periods of depression. Parents noticed that their children seemed caring and understanding of illness, but were also anxious when the parent was ill or in hospital, and reacted with anger or frustration to restrictions in social activities. Strategies commonly used to deal with difficulties were turning to family for support and trying to control symptoms. Support from health professionals was variable. Parents wanted more practical assistance, information for families on the effects of IBD, and support in coping with the condition. Awareness-raising about IBD was considered important within health, social services, education and housing departments. CONCLUSIONS: As a group in regular contact with parents with IBD, health professionals have an important role to play in providing support. Staff should be particularly vigilant towards parents with severe symptoms, mothers of younger children, and those receiving limited help from their families. PMID- 11872104 TI - Being in a fragmented and isolated world: interviews with carers working with a person with a severe autistic disorder. AB - AIM: To illuminate the meaning of being a carer for a person with a severe autistic disorder. BACKGROUND: Carers working with people with severe autism are occasionally exposed to residents' self-injurious behaviours and violent actions and at time residents appear resistant to all forms of treatment. DESIGN/METHOD: A qualitative case study was conducted. Six Swedish carers enrolled nurses (ENs), working on a special ward in a nursing home were interviewed about their lived experiences when caring for an individual with a severe autistic disorder. Narrative interviews were conducted and interpreted using a phenomenological hermeneutic method inspired by Paul Ricoeur. FINDINGS: Two themes were formulated which describe the carers' reality and their dream of an ideal. This ideal described carers' experiences of being trapped in a segmented and isolated care reality and their longing to achieve a sense of wholeness. The findings were interpreted and reflected on in the light of a framework inspired by the German philosopher Karl Jaspers in order to achieve a deeper understanding of the text. CONCLUSIONS: In their desperation, the carers used their empirical knowledge based on scientific knowledge, which could be understood as a substitute for their vision of a consolating wholeness. This paper shows that searching for a substitute to consolation seems to be an important aspect of the meaning of being a carer for a person with a severe autistic disorder. PMID- 11872106 TI - The short-term effects of myofascial trigger point massage therapy on cardiac autonomic tone in healthy subjects. AB - AIM OF THE STUDY: To investigate the effects of myofascial trigger-point massage therapy to the head, neck and shoulder areas on cardiac autonomic tone.Background. No studies have reported on the effect of back massage on autonomic tone as measured by heart rate variability. This is especially relevant to the nursing profession, as massage is increasingly available as a therapy complementary to conventional nursing practice. DESIGN/METHODS: An experimental study in which subjects were initially placed in age- and sex-matched groups and then randomized to treatment or control by alternate allocation. The study involved 30 healthy subjects (16 female and 14 male, aged 32.47 +/- 1.55 years, mean +/- standard error). A 5-minute cardiac interbeat interval recording, systolic and diastolic blood pressure and subjective self-evaluations of muscle tension and emotional state were taken before and after intervention. Autonomic function was measured using time and frequency domain analysis of heart rate variability. RESULTS: Following myofascial trigger-point massage therapy there was a significant decrease in heart rate (P < 0.01), systolic blood pressure (P=0.02) and diastolic blood pressure (P < 0.01). Analysis of heart rate variability revealed a significant increase in parasympathetic activity (P < 0.01) following myofascial trigger-point massage therapy. Additionally both muscle tension and emotional state, showed significant improvement (P < 0.01). CONCLUSIONS: In normal healthy subjects myofascial trigger-point massage therapy to the head, neck and shoulder areas is effective in increasing cardiac parasympathetic activity and improving measures of relaxation. PMID- 11872108 TI - The application of vignettes in social and nursing research. AB - AIM: The aim of this paper is to review the potential for, and the limitations of, the use of vignettes in research that seeks an understanding of people's attitudes, perceptions and beliefs, particularly with regard to sensitive subjects such as health care. BACKGROUND: Vignettes, in the form of text or pictures presented to research participants to prompt responses to interview questions, are widely used throughout the social sciences although their use in nursing research is less developed. REVIEW FOCUS: This review paper begins by addressing the differences between vignettes and real life processes. The following sections explore some of the practical advantages and pitfalls of using vignettes in social and nursing research. CONCLUSION: The paper demonstrates how vignettes can be very useful research tools yielding valuable data when studying people's attitudes, perceptions and beliefs in social and nursing research. PMID- 11872107 TI - Nursing diagnosis taxonomy across the Atlantic Ocean: congruence between nurses' charting and the NANDA taxonomy. AB - PURPOSE AND AIMS: The purpose of this study was to analyse expressions or terms used by nurses in Iceland to describe patient problems. The classification of NANDA was used as reference. The research questions were: (a) Does NANDA terminology represent patient problems documented by Icelandic nurses? (b) If so, what kind of nursing diagnoses does it represent? (c) What kind of patient problems are not represented by NANDA terminology? (d) What are the most frequent nursing diagnoses used? METHODS: A retrospective chart review was conducted in a 400 bed acute care hospital in Iceland. The sample was defined as nursing diagnosis statements in charts of patients hospitalized in two 6-month periods in two separate years. The data were analysed according to a predefined grading system based on the PES format or Problem -- (A)aetiology -- Signs and symptoms. RESULTS: A total of 1217 charts were used for the study, which yielded 2171 nursing diagnoses statements for analysis. Charts with at least one nursing diagnosis documented were 60.1% and the number of diagnoses per patient ranged from 0 to 10, with 65% of charts with three diagnoses or less. The number of diagnoses correlated with patients' length of stay, but not with increased age of the patients. The average number of statements per patient was 3.28. Almost 60% of the diagnoses were according to NANDA terminology, another 20% were stated as procedures, medical diagnoses or risks for complications. The 20 most frequently used nursing diagnoses accounted for 80% of all diagnoses documented. Discrepancy between nurses' documentation on emotional problems and availability of diagnosis in the NANDA taxonomy was evident. CONCLUSION: It can be concluded that the NANDA taxonomy seems to be culturally relevant for nurses in different cultures. PMID- 11872109 TI - A method comparison study to assess the reliability of urine collection pads as a means of obtaining urine specimens from non-toilet-trained children for microbiological examination. AB - BACKGROUND: In young, non-toilet-trained children, the collection of a urine sample for microbiology can be challenging, with the application of a urine bag being the main method of collection. However, recent research has shown that absorbent pads can be used yielding similar results to bag specimens. However, weaknesses in study design erode confidence in research findings. Therefore, improvements in research design are required to fully evaluate the reliability of pad collection. AIMS OF THE STUDY: This pilot study sought to test the feasibility of a technique for the collection of concurrent bag/pad urine samples from non-toilet-trained children, and to assess the reliability of urine pads over bags as a collection method for urine specimens for microbiological evaluation. DESIGN: A pilot, method comparison study. METHODS: Twenty concurrent bag and pad specimens were collected from non-toilet-trained children, following parental consent. Urine specimens were analysed for presence or absence of white cell count (WBC), and bacterial growth, using standard laboratory methods. DATA ANALYSIS: The Kappa (kappa) statistics and confidence interval (CI) estimation were used to assess agreement between the two collection methods. RESULTS: Despite concurrent samples there was a lack of agreement between bag and pad specimens on both main outcome measures. Agreement between bag and pad specimens for the presence of WBC yielded a kappa=0.10 (95% CI: 0.19, 0.39), indicating poor agreement, while a kappa of 0.5 (95% CI: 0.12, 0.88) was calculated for the degree of agreement in bacterial growth reflecting moderate agreement. Differences in proportions of the presence of WBC between bag and pad did not quite reach significance at the 5% level 0.2 (95% CI: 0.00, 0.42, P=0.062). For cultures the difference was calculated as 0.15 (95% CI: 0.05, 0.35, P=0.125). CONCLUSION: The pilot study demonstrates that concurrent urine samples can be obtained without difficulty. Despite poor to moderate agreement on outcome measures the level of agreement is greater than reported in those other studies, that use non-current methods of urine collection, suggesting an advantage of the concurrent technique. It is recommended that larger scale studies be undertaken using the concurrent collection technique to assess reliability of these findings. PMID- 11872110 TI - The content of management of violence policy documents in United Kingdom acute inpatient mental health services. AB - AIM OF THE STUDY: The aim of the study was to examine the content of Trust policies concerning the prevention and management of violence in acute in-patient settings in order to establish their usefulness as guidance for staff in this difficult, complex, and controversial aspect of inpatient psychiatric care. BACKGROUND: Violence is a commonly encountered problem in inpatient psychiatric settings. There are legal requirements for workplaces in general and mental health care facilities in particular to develop safe systems of work based upon the findings of assessments of this risk. Policies have a key role to play in making explicit the responsibilities of both employer and employees, and specifying standards of acceptable practice. DESIGN: A cross-sectional survey methodology was used, which entailed examination of the content of management of violence policies that had been forwarded to the authors from 40 Trusts providing acute inpatient psychiatric care throughout England, Scotland, Wales, and Northern Ireland. FINDINGS: Policies were found to vary widely in their content, and serious shortcomings were noted in the extent to which policies included information regarding their status and review, advice on the prevention of violence, the management of violent incidents, and postincident action. CONCLUSIONS: Further research is needed to tease out the extent to policies which are lacking in content, reflect shortcomings in the organizational approach to the prevention and management of violence by Trusts, and the extent to which such shortcomings result in harm being suffered by staff and/or patients. An alternative format for the presentation of management of violence policies is discussed, and items that should be included in inpatient units' management of violence policies are suggested. PMID- 11872112 TI - A role for intestinal mycoplasmas in the aetiology of Crohn's disease? PMID- 11872115 TI - Acid stress in the food pathogen Bacillus cereus. AB - AIMS: The pathogen Bacillus cereus, which is associated with a number of foods including dairy products, was studied for its response to acid stress during the exponential phase. METHODS AND RESULTS: Bacillus cereus was found to adapt to acid stress (pH 4.6) when pre-exposed to a non-lethal, inducing pH of 6.3 or to inducing concentrations of heat, ethanol, salt or hydrogen peroxide. Cells were found to maintain their internal pH at a higher level than the external acid pH and adapted cells had a higher internal pH than unadapted cells. A constitutive acid-sensitive mutant that was also heat- and ethanol-sensitive was found to be capable of high levels of adaptation despite its lack of induction of proteins induced in the wild type by exposure to moderate pH (6.3) values. CONCLUSIONS: A number of proteins were found to be underexpressed in the mutant compared with the wild type at pH 6.3, including some with homology to ribosomal proteins and to the sporulation regulator RapK, while one differentially expressed band contained two proteins, one of which was homologous to the competence regulator CodY. SIGNIFICANCE AND IMPACT OF THE STUDY: The work has implications for the processing of B. cereus-associated foods by acidification. The linked developmental processes of stationary phase, sporulation and possibly competence appear to be involved in the response to acid stress. PMID- 11872114 TI - Inhibition of the adherence of Escherichia coli strains to basement membrane by Lactobacillus crispatus expressing an S-layer. AB - AIMS: This study aimed to evaluate the efficiency with which Lactobacillus crispatus JCM 5810 inhibited the adhesion of enteric pathogens to a synthetic basement membrane and to elucidate the mechanism underlying the inhibition. METHODS AND RESULTS: Lactobacillus crispatus JCM 5810 inhibited the adhesion of three diarrhoeagenic Escherichia coli strains to a reconstituted basement membrane preparation called Matrigel, used as a model of a damaged intestinal tissue site. Inhibition was also observed with the use of immobilized laminin, a major component of Matrigel, but diminished after the removal of S-layer protein (CbsA) from JCM 5810 cells. The isolated CbsA inhibited the adhesion of E. coli to both Matrigel and immobilized laminin. Lactobacillus crispatus JCM 5810 and CbsA seem to inhibit pathogenic E. coli from adhering to basement membrane via competition with laminin molecules for binding sites. CONCLUSIONS: These results suggested that not only Lact. crispatus JCM 5810 cells but CbsA alone might prevent pathogens from colonizing damaged intestinal tissues. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first study to show the applied aspect of Lactobacillus S-layer protein. PMID- 11872113 TI - Combined effects of water activity, temperature and chemical treatments on the survival of Salmonella and Escherichia coli O157:H7 on alfalfa seeds. AB - AIMS: The objective of this study was to determine the combined effects of water activity (a(w)), chemical treatment and temperature on Salmonella and Escherichia coli O157:H7 inoculated onto alfalfa seeds. METHODS AND RESULTS: Alfalfa seeds inoculated with Salmonella or E. coli O157:H7 and adjusted to various a(w) values were subjected to simultaneous and separate treatments with chemicals and heat. The rate of death of both pathogens was correlated with increased a(w) (0.15 0.60) and temperature (5-37 degrees C) over a 52-week storage period. Higher seed a(w) enhanced the inactivation of pathogens on seeds heated at 50-70 degrees C for up to 24 h. Treatment of seeds with water, 1% Ca(OH)2, 1% Tween 80, 1% Ca(OH)2 plus 1% Tween 80 or 40 mg l(-1) Tsunami 200 at 23 or 55 degrees C for 2 min significantly (alpha=0.05) reduced populations of Salmonella and E. coli O157:H7. CONCLUSIONS: Overall, at the combinations of temperature and concentrations of chemicals tested, 1% Ca(OH)2 was most effective in killing Salmonella and E. coli O157:H7 without reducing seed viability. SIGNIFICANCE AND IMPACT OF THE STUDY: None of the treatments evaluated in this study, whether applied separately or in combination, eliminated Salmonella or E. coli O157:H7 on alfalfa seeds without sacrificing the viability of the seeds. It remains essential that practices to prevent the contamination of alfalfa seeds be strictly followed in order to minimize the risk of Salmonella and E. coli O157:H7 infections associated with sprouts produced from these seeds. PMID- 11872116 TI - Changes in activity of extracellular enzymes in dual cultures of Lentinula edodes and mycoparasitic Trichoderma strains. AB - AIMS: The main problem that arises during the cultivation of Lentinula edodes, the Asian Shiitake mushroom, is that the logs on which the cultivation is performed are contaminated by competing micro-organisms, especially Trichoderma spp. The aim of this study was to examine the changes in activity of extracellular enzymes in dual cultures of Trichoderma spp. and L. edodes. METHODS AND RESULTS: Extracellular enzyme activities were determined spectrophotometrically. Trichoderma enzymes important for the degradation of fungal cell walls (N-acetyl-beta-glucosaminidase and laminarinase) were shown to be induced by inactive L. edodes mycelia in liquid culture. The changes that occurred in the extracellular enzyme activities of L. edodes and mycoparasitic Trichoderma spp. (T. aureoviride, T. harzianum and T. viride) were examined during antagonistic interactions on solid medium. The extracellular enzyme patterns of both partners proved to be altered. Trichoderma spp. were induced to produce N-acetyl-beta-glucosaminidase and laminarinase in the presence of active L. edodes mycelia, similarly as observed in liquid culture. The activities of both laccase and manganese peroxidase of L. edodes decreased after physical contact with active Trichoderma mycelia, possibly in consequence of the beginning of degradation of L. edodes by the Trichoderma enzymes. However, besides a decrease in manganese peroxidase activity, an enhancement of L. edodes laccase activity was observed on solid media containing crude culture fluids from Trichoderma liquid cultures. The metabolites responsible for these effects proved to be heat stable. CONCLUSIONS: Induction and inhibition of several extracellular enzymes of both partners were shown in dual cultures of L. edodes and Trichoderma strains, indicating the important role of these enzymes in the antagonistic interaction between the two species. SIGNIFICANCE AND IMPACT OF THE STUDY: As the main problem during the large-scale cultivation of L. edodes is the contamination of the growth substrate by Trichoderma mycelia, the particular knowledge of the mechanism of this competition might be relevant. PMID- 11872118 TI - Culturability, injury and morphological dynamics of thermophilic Campylobacter spp. within a laboratory-based aquatic model system. AB - AIMS: To study the survival processes of thermophilic Campylobacter spp. within a modelled aquatic system and particularly the involvement and survival potential of viable but non-culturable forms. METHODS AND RESULTS: The survival and morphological characteristics of populations of thermophilic Campylobacter species exposed to simulated aquatic conditions were examined using a combination of cultural and microscopic techniques. Populations underwent progressive decay when exposed to simulated aquatic conditions. The rates of population decay were observed to be significantly greater at the higher temperature (20 degrees C) with a rapid transition of the dominant sub-populations from non-stressed to dead cells occurred within 3 days. At 10 degrees C the rate of culturability loss was much reduced with substantial development (approx. 80% of total population) of viable but non-culturable (VBNC) populations by all species within 3 days, declining to represent approximately 5-25% of the total population at day 60. Significant differences (P < 0.001) were identified between decay rates as a consequence of different species, sub-populations and temperature but not between sub-populations of different species. Morphological variants including spiral, elongated spirals and rods, short rods and coccoid forms were identified. The endpoints of morphological transition were temperature-independent and isolate specific yet the rate of morphological transition was directly related to temperature and approximately equivalent between species. CONCLUSION: The VBNC state is a transitory stage in the degeneration of Campylobacter population within the aquatic environments simulated during this study. SIGNIFICANCE AND IMPACT OF THE STUDY: VBNC cells form the most persistent, viable, potentially pathogenic sub-population of Campylobacter populations exposed to aquatic stress conditions. PMID- 11872117 TI - Sources of Salmonella on broiler carcasses during transportation and processing: modes of contamination and methods of control. AB - AIMS: The prevalence and types of salmonella in broiler chickens during transportation and during slaughter and dressing were studied. This was part of a comprehensive investigation of salmonellas in two UK poultry companies, which aimed to find the origins and mechanisms of salmonella contamination. METHODS AND RESULTS: Salmonellas were isolated using cultural methods. Serovars of Salmonella detected during rearing were usually also found in a small proportion of birds on the day of slaughter and on the carcasses at various points during processing. There was little evidence of salmonellas spreading to large numbers of carcasses during processing. Many serovars found in the feedmills or hatcheries were also detected in the birds during rearing and/or slaughter. Transport crates were contaminated with salmonellas after washing and disinfection. CONCLUSIONS: Prevalence of salmonellas fell in the two companies during this survey. A small number of serovars predominated in the processing plants of each company. These serovars originated from the feed mills. Reasons for transport crate contamination were: (1) inadequate cleaning, resulting in residual faecal soiling; (2) disinfectant concentration and temperature of disinfectant too low; (3) contaminated recycled flume water used to soak the crates. SIGNIFICANCE AND IMPACT OF THE STUDY: Efforts to control salmonella infection in broilers need to concentrate on crate cleaning and disinfection and hygiene in the feed mills. PMID- 11872119 TI - Bacteria in heart and lungs of young chicks. AB - AIMS: The purpose of these experiments was to determine whether the heart and lungs of young chicks harboured bacteria. METHODS AND RESULTS: Samples of the heart and lungs were aseptically removed from chicks on scheduled sampling days. Experiment 1 showed that of the 360 birds evaluated during the late embryonic and early post-hatching periods, only 10.8% harboured bacteria in the heart, lungs, and heart and lungs simultaneously. Experiment 2 suggested that bacteria in these organs were transient. Twenty-three bacterial species were found in the hearts whereas 30 were found in the lungs. Experiment 3 showed that only 1.4% of embryos harboured bacteria in the yolk, albumen, heart and lungs whereas 12.9% of the embryos had bacteria in the air cell. CONCLUSIONS: During the post-hatching period, there was a higher incidence of bacterial isolation in the heart and lungs, whilst during the embryonic development period, there was a lower incidence of bacterial isolation from these two organs. Results suggested that the heart and lungs do not have a residual bacterial flora; rather, opportunistic bacteria occasionally pass through these tissues. SIGNIFICANCE AND IMPACT OF THE STUDY: These experiments proved that bacteria could be isolated in the heart and lungs of healthy chicks reared from E17 to 3 weeks of age. PMID- 11872120 TI - Genetic diversity of vaginal lactobacilli from women in different countries based on 16S rRNA gene sequences. AB - AIMS: Lactobacilli are widely distributed in food and the environment, and some colonize the human body as commensal bacteria. The aim of this study was to determine the species of lactobacilli that colonize the vagina and compare them with those found in food and the environment. METHODS AND RESULTS: Thirty-five Lactobacillus strains from women from seven countries were isolated, and sequences from 16S rRNA genes were determined and compared with existing data in GenBank. A phylogenetic tree was achieved using the Neighbour-Joining method based on the analysis of 1465 nucleotides. The results showed that most vaginal isolates were L. crispatus, L. jensenii and L. gasseri. Some were L. vaginalis, L. fermentum, L. mucosae, L. paracasei and L. rhamnosus. Two isolates from a native American woman displayed distinct branches, indicating novel phylotypes. Few vaginal isolates matched food or environmental Lactobacillus species. CONCLUSIONS: Most women worldwide were colonized by three common Lactobacillus species: L. crispatus, L. jensenii and L. gasseri. SIGNIFICANCE AND IMPACT OF THE STUDY: Knowledge of vaginal Lactobacillus species richness and distribution in women worldwide may lead to the design of better probiotic products as bacterial replacement therapy. PMID- 11872121 TI - Effects of depuration of molluscs experimentally contaminated with Escherichia coli, Vibrio cholerae 01 and Vibrio parahaemolyticus. AB - AIMS: The aim of the present study was to investigate the behaviour of two pathogenic vibrios (Vibrio cholerae O1 and Vibrio parahaemolyticus) during depuration and to compare it with that of Escherichia coli, used as an indicator of suitability for consumption. METHODS AND RESULTS: Samples of Mytilus galloprovincialis were experimentally contaminated with E. coli, V. cholerae O1 and V. parahaemolyticus, depurated in a pilot plant using ozone and analysed at selected intervals. Numbers of E. coli and vibrios were estimated using an MPN method. The presence of vibrios was confirmed by the use of PCR. The target genes used were ctx for V. cholerae O1 and the restriction fragment pR72H for V. parahaemolyticus. There was a substantially smaller reduction in the numbers of both vibrios (approximately 1 log) during the depuration process than of E. coli (approximately 3 log). CONCLUSIONS: The results confirm the inadequacy of E. coli as an indicator that molluscs have been cleansed of other microbiological agents. SIGNIFICANCE AND IMPACT OF THE STUDY: The study confirms the risk associated with the consumption of mussels and the need to correctly conserve and cook them prior to consumption. PMID- 11872122 TI - Diversity of Bacillus thuringiensis strains from Colombia with insecticidal activity against Spodoptera frugiperda (Lepidoptera:Noctuidae). AB - AIMS: To identify and characterize Bacillus thuringiensis strains highly toxic to Spodoptera frugiperda, and to explore the genetic diversity of such strains. METHODS AND RESULTS: The insecticidal activity of 1100 strains of B. thuringiensis from Colombian soil samples was assayed against first instar S. frugiperda larvae, and 32 active strains were found. After a second bioassay evaluation, the eight most potent strains were selected for further characterization, which included crystal protein profiles determined by polyacrylamide gel electrophoresis, plasmid profile, plasmid restriction patterns, cry gene composition, qualitative determination of beta-exotoxin production, random amplified polymorphic DNA, serotyping, and toxicity to S. frugiperda. All Colombian strains contained cry1Aa, cry1Ab, cry1Ac, cry1B, cry1C and cry1D genes. However, PCR profiles of the Colombian strains suggested the presence of variants of the cry1 genes. Serotyping indicated that these strains belong to the kurstaki, thuringiensis, canadiensis and indiana subspecies. Interestingly, three strains belonging to different serotypes and subspecies were found in the same soil sample, and toxicity ranged between 11 and 976 ng cm(-2) of diet. CONCLUSIONS: It has been shown that B. thuringiensis strains belonging to different serotypes and displaying variable potency to S. frugiperda larvae can be found in the same soil sample. SIGNIFICANCE AND IMPACT OF THE STUDY: The results obtained indicate that some of the B. thuringiensis strains studied could be of interest for further development for S. frugiperda control programmes. PMID- 11872123 TI - Cytometric monitoring of growth, sporogenesis and spore cell sorting in Paenibacillus polymyxa (formerly Bacillus polymyxa). AB - AIMS: Formation of bacterial endospores is a basic process in Gram-positive bacteria and has implications for health, industry and the environment. Flow cytometry offers a practical alternative for the rapid detection, enumeration and characterization of bacterial endospores. METHODS AND RESULTS: Paenibacillus polymyxa was chosen for this study because its spores cause sporangium deformation and have thick walls with a star-shaped section. Sporulating populations were analysed with a particle analyser and a flow cytometer after labelling with propidium iodide and Syto-13. Flow cytometric detection of single spores was confirmed by optical and scanning electron microscopy after cell sorting. Four cell sub-populations were cytometrically detected in P. polymyxa cultures grown in liquid sporulation medium. Two sub-populations consisted of vegetative cells differing in both morphology and viability; the other two sub populations consisted of spores differing in their viability. CONCLUSIONS: This work has shown that flow cytometry is a simple and fast method (less than 15 minutes for sample preparation and analysis) for the study of the sporulation in P. polymyxa. The use of this technique allowed both detection and quantification of sporulation inside a culture, and distinguished cells that differed in viability despite being morphologically identical under microscopic observation. SIGNIFICANCE AND IMPACT OF THE STUDY: Flow cytometry has been proved to be a valuable tool for the analysis of sporulation in P. polymyxa cultures, with the unique capacity of distinguishing between endospores and vegetative cells, and between live and dead cells, in the same analysis. An important percentage of non viable endospores has been found in aged cultures using this method. PMID- 11872124 TI - Proteolytic activity of Staphylococcus xylosus strains on pork myofibrillar and sarcoplasmic proteins and use of selected strains in the production of "Naples type" salami. AB - AIMS: The aim of this study was to determine the proteolytic activities of Staphylococcus xylosus strains on sarcoplasmic and myofibrillar proteins in order to evaluate the suitability of selected strains as starter cultures in the processing of a dry fermented pork sausage. METHODS AND RESULTS: The proteolytic activity of 27 strains of Staphylococcus xylosus on sarcoplasmic and myofibrillar proteins was determined by agar plate method, o-phtaldialdehyde (OPA) spectrophotometric assay and sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). Four strains were selected for the formulation of six starter cultures to use in the production of "Naples type" salami. The proteolytic contribution of starters was determined by SDS-PAGE, comparing the protein profile of inoculated sausages with that of uninoculated sausages after 0, 15 and 33 days of ripening. The results showed that the proteolytic activity of some strains, determined by the agar plate method, were not confirmed by electrophoretic and spectrophotometric assays. In fact, of 24 strains of Staphylococcus xylosus able to hydrolyse muscle protein extracts on agar plate, only 12 strains were shown to change SDS-PAGE profile of pork proteins. The SDS PAGE profile of sarcoplasmic proteins extracted from all sausages showed that the major changes were produced with starters S3, S4 and S5 after 15 days of ripening. Also myofibrillar proteins undergo major changes after 15 days of ripening and the protein profiles showed the same pattern in all samples, except for the sausages produced with starter S4. CONCLUSIONS: The results of this work showed that the muscle protein extracts hydrolysis test is suitable for preliminary screening of Staphylococcus xylosus strains on the basis of their proteolytic activity. However, evaluation of muscle protein hydrolysis in a food model system could then be more appropriate for selecting micro-organisms for use as starter cultures for fermented sausages. SIGNIFICANCE AND IMPACT OF THE STUDY: The potential of the findings is discussed with reference to the formulation of starter cultures for the dry fermented sausages production. PMID- 11872125 TI - Genetic diversity among Geotrichum candidum strains from various substrates studied using RAM and RAPD-PCR. AB - AIMS: Assessment of genetic diversity within the species Geotrichum candidum and development of tools to trace the strains that play an important role in the agro food industry. METHODS AND RESULTS: RAM-PCR and RAPD-PCR techniques were assessed for their ability to discriminate 57 strains of various morphotypes, substrates and geographical origin. The techniques were complementary and, when combined, allowed us to discriminate isolates. Moreover, we established a link between a taxon and its occupation of an ecological niche, which should not be confused with the substrate of isolation. CONCLUSIONS: We observed a high degree of diversity, which could be linked to the variety of the ecological niches chosen and to the high degree of morphological polymorphism encountered within the species. SIGNIFICANCE AND IMPACT OF THE STUDY: Used in combination, RAM-PCR and RAPD-PCR permit traceability and monitoring systems for G. candidum strains during food processing. PMID- 11872126 TI - Phenotypic diversity of Campylobacter isolates from sporadic cases of human enteritis in the UK. AB - AIMS: The aim of this study was to identify and subtype a large collection of isolates of Campylobacter spp. to quantify diversity among strains causing human disease from geographically diverse sources in the United Kingdom. METHODS AND RESULTS: Isolates were characterized by the Penner serotyping scheme, Preston phage typing and biotyping methods. The diversity index calculated from the combined results of all three methods was 0.997 and indicated that isolates from sporadic cases of infection are very diverse. Strong associations between common phagetypes (PG52, PG121 and PG55) and the three most common serotypes (HS1, HS2 and HS4) found in the study were evident. CONCLUSIONS: Strains of C. jejuni causing human infections in the United Kingdom are very phenotypically diverse. Individual strains characterized by serotype, phagetype and biotype were detected throughout the 7-month study period and from geographically distinct sources, indicating an unrecognized outbreak or other epidemiologically significant source of human infection. SIGNIFICANCE AND IMPACT OF THE STUDY: The low frequency incidence of most C. jejuni strains should enable easy recognition of outbreaks by strain type surveillance at local, regional and national level in the United Kingdom. The characterization of common strain profiles in this study by simple phenotypic methods could provide the basis for strain specific epidemiological studies for reservoirs of infection and transmission routes for human infection. PMID- 11872127 TI - Development and application of a nested PCR to monitor brood stock salmonid ovarian fluid and spleen for detection of the fish pathogen Flavobacterium psychrophilum. AB - AIMS: To develop a nested PCR to detect Flavobacterium psychrophilum based on the intergenic spacer region 16S-23S rRNA and in 16S rRNA for analysis of brood stock salmonid fish samples. METHODS AND RESULTS: The sensitivity and specificity of the test was evaluated using pure cultures, spiked and naturally contaminated samples. Samples were internal organs (spleen and kidney), eggs and ovarian fluid from rainbow trout and coho salmon from European fish farms (France, Spain). This nested PCR was more specific and sensitive that the nested PCR based on 16S rRNA sequences primers only. The detection limit of this PCR assay was one bacterium per PCR tube corresponding to 10 bacteria/mg of spleen and 5 bacteria/ml from ovarian fluid. Analysis of mixed ovarian fluid samples from reproductive salmonids in various French hatcheries demonstrated that 69% of hatcheries were contaminated with Fl. psychrophilum. The analysis of individual samples demonstrated that 39% of rainbow trout (Oncorhynchus mykiss) and 62.5% of coho salmon (O. kisutch) samples were contaminated. CONCLUSIONS: The results demonstrated a very sensitive and specific detection of this fish pathogen and that most of the female rainbow trout and coho salmon breeders analysed carry Fl. psychrophilum in the ovarian fluid. SIGNIFICANCE AND IMPACT OF THE STUDY: The understanding of Fl. psychrophilum dissemination and transmission and the detection of asymptomatic carriers is important for the development of free breeders stock and for significantly decreasing Flavobacteriose. PMID- 11872128 TI - Pyrimidine biosynthesis in Pseudomonas oleovorans. AB - AIMS: To investigate the regulation of de novo pyrimidine biosynthesis in the polyhydroxyalkanoate-producing bacterium Pseudomonas oleovorans at the level of enzyme synthesis and at the level of aspartate transcarbamoylase activity. METHODS AND RESULTS: The effect of pyrimidine supplementation on the pyrimidine biosynthetic pathway enzyme activities was analysed relative to carbon source. Two uracil auxotrophs of P. oleovorans were isolated that were deficient for aspartate transcarbamoylase or dihydroorotase activity. Pyrimidine limitation of these auxotrophs increased the de novo pathway activities to varying degrees depending on the pathway mutation and the carbon source utilized. At the level of aspartate transcarbamoylase activity, pyrophosphate and uridine ribonucleotides were found to be strongly inhibitory of the Ps. oleovorans enzyme. CONCLUSIONS: Pyrimidine biosynthesis is regulated in Ps. oleovorans. Taxonomically, the regulation of the pyrimidine biosynthetic pathway appeared dissimilar from previously studied Pseudomonas species. SIGNIFICANCE AND IMPACT OF THE STUDY: New insights regarding the regulation of nucleic acid metabolism are provided that could prove significant during the genetic manipulation of Ps. oleovorans to increase the synthesis of polyhydroxyalkanoates. PMID- 11872129 TI - The detection of Bifidobacterium adolescentis by colony hybridization as an indicator of human faecal pollution. AB - AIMS: To develop an improved method for the detection of Bifidobacterium adolescentis as an indicator of human faecal pollution. METHODS AND RESULTS: Bifidobacterium medium (BFM) was identified as the optimal medium for the recovery of bifidobacteria from human effluent. Dilutions of faeces and effluent from both humans and animals were filtered, grown on BFM and human specific B. adolescentis identified via colony hybridization with a digoxigenin (DIG) labelled oligonucleotide probe. CONCLUSIONS: The combination of BFM with colony probing allows the detection of B. adolescentis, a specific indicator of human faecal pollution. SIGNIFICANCE AND IMPACT OF THE STUDY: It is now technically feasible to use B. adolescentis as indicators of human faecal pollution, and studies to examine the survival and appropriateness of bifidobacteria in this role can be initiated. PMID- 11872130 TI - Isolation and identification of synthetic pyrethroid-degrading bacteria. AB - AIMS: To isolate, select, identify and assess the potential for the biodegradation of synthetic pyrethroids (SPs) in sheep dips. METHODS AND RESULTS: SP-degrading bacteria were isolated from a mixed soil sample consisting of garden soil and soils from farms where SPs had been used. The two largest in size were then identified using microscopy, biochemical and genetic techniques to be members of the genera Pseudomonas and Serratia. By comparing the 16S rRNA gene sequences, the Pseudomonas sp. discovered was shown to group within the Pseudomonas fluorescens intrageneric cluster. The Serratia isolated was closely related to Serratia plymuthica. Cell growth and degradation was greatest in the Pseudomonas sp. culture where there was breakdown of 60 mg l(-1) to 6 mg l(-1) technical cypermethrin in 20 days. Tolerance to the SPs was greater in the Pseudomonas sp. but was found to depend on the availability of other carbon sources and nutrients. CONCLUSIONS: The bacteria characterized show the potential to be used in a bioremediation application for the treatment of SP residues. SIGNIFICANCE AND IMPACT OF THE STUDY: The SP-degrading bacteria may have use in the disposal of used SP residues and with further research could lead to an alternative route of disposal for use in agriculture or industry. PMID- 11872131 TI - Enrichment of a microbial culture capable of degrading endosulphate, the toxic metabolite of endosulfan. AB - AIMS: The aim of this study was to isolate a source of enzymes capable of degrading endosulphate (endosulfan sulphate), the toxic metabolite of the pesticide endosulfan. METHODS AND RESULTS: A microbial broth culture capable of degrading endosulphate was enriched from endosulfan-contaminated soil by providing the metabolite as the sole source of sulphur in broth culture. No microbial growth was observed in the absence of endosulphate. In the presence of endosulphate, growth of the culture occurred with the concomitant formation of three chlorine-containing compounds. Thin layer chromatography and gas chromatography--mass spectral analysis identified these metabolites as endosulfan monoaldehyde, 1,2,3,4,7,7-hexachloro-5,6-bis(methylene)bicyclo[2.2.1]-2-heptene and 1,2,3,4,7,7-hexachloro-5-hydroxymethylene-6-methylenebicyclo[2.2.1]-2 heptene. The second and third compounds have not been reported in previous metabolic studies. The enriched culture was also able to utilize alpha- and beta endosulfan as sulphur sources, each producing the hydrolysis product endosulfan monoaldehyde as the sole chlorine-containing metabolite. Alpha-endosulfan was more readily hydrolysed than the beta-isomer. CONCLUSIONS: This study isolated a mixed microbial culture capable of degrading endosulphate. The products of degradation were characterized as novel endosulfan metabolites. SIGNIFICANCE AND IMPACT OF THE STUDY: This study describes the isolation of a mixed microbial culture that is potentially a valuable source of hydrolysing enzymes for use in enzymatic bioremediation, particularly of endosulphate and alpha-endosulfan residues. PMID- 11872132 TI - A quantitative study of the survival of two species of Candida on porous and non porous environmental surfaces and hands. AB - AIMS: In spite of the importance of many species of Candida as human pathogens, little is known about their ability to survive on animate and inanimate surfaces. Such information is essential in understanding the vehicles and modes of their spread, and in designing proper infection control strategies against them. The aim of this study was to generate comparative quantitative data in this regard. METHODS AND RESULTS: The survival of one clinical isolate each of Candida albicans and C. parapsilosis on two types of hard inanimate surfaces (glass and stainless steel) and two types of fabrics (100% cotton and a blend of 50% cotton and 50% polyester) was evaluated under ambient conditions (air temperature 22 +/- 2 degrees C; relative humidity 45-62%) using quantitative test protocols. The survival of C. albicans was also assessed on human skin, using the fingerpads of adult volunteers as carriers. Each carrier surface received 10 microl of the test suspension containing a soil load to simulate body fluids. When dried on glass and stainless steel carriers, C. albicans and C. parapsilosis remained viable for at least three and 14 days, respectively. Both species could survive for at least 14 days on both types of fabric. On the skin, 20% of the viable C. albicans remained detectable one hour post-inoculation. SIGNIFICANCE AND IMPACT OF THE STUDY: This quantitative and comparative study demonstrated the potential for, and differences in the ability of clinically significant species of Candida to remain viable on porous and non-porous inanimate surfaces as well as on human hands. These results should help in understanding the epidemiology of nosocomial infections due to Candida, and in designing better prevention and control strategies against them. PMID- 11872133 TI - Growth, morphology and surface properties of Listeria monocytogenes Scott A and LO28 under saline and acid environments. AB - AIMS: The effect of salt and acid on the growth and surface properties of two strains of Listeria monocytogenes was investigated. METHODS AND RESULTS: Medium acidification and NaCl supplementation induced a marked increase in the lag and growth times (up to fivefold higher) and a decrease in the maximal optical density. Due to a strong synergic effect of pH and NaCl, growth was only detected after 280 h incubation for Scott A and not detected after 600 h for LO28 at pH 5.0 and 10% NaCl. Furthermore, the addition of NaCl in acidic conditions gave rise to cell filamentation and cell surfaces became strongly hydrophilic. CONCLUSIONS: Some L. monocytogenes strains subjected to high NaCl concentrations in acidic conditions are able to grow but may present altered adhesion properties due to modification of their surface properties. SIGNIFICANCE AND IMPACT OF THE STUDY: This study highlighted that L. monocytogenes do represent a hazard in acid and salted foods, such as soft cheese. PMID- 11872134 TI - Mechanisms contributing to hypochlorous acid resistance of a Salmonella isolate from a poultry-processing plant. AB - AIMS: We have recently reported the isolation of Salmonella that have acquired tolerance to hypochlorous acid (HOCl) (Mokgatla et al. 1998). The aim of this work was to investigate possible protective mechanisms involved in the increased tolerance to HOCl of a selected resistant strain. METHODS AND RESULTS: One resistant (Salmonella 104) and one sensitive (Salmonella 81) isolate in exponential phase were exposed to HOCl at a final active concentration of 28 mg l(-1). Cultures were assayed for superoxide dismutase and catalase activity, as well as for four membrane-bound dehydrogenases (malate, lactate, glutamate and glucose-6-phosphate dehydrogenase). The degree of single-strand breaks in genomic DNA was analysed and lipopolysaccharide profiles determined. The resistant Salmonella isolate differed from the sensitive isolate in a number of ways. It responded within 10 min of exposure by producing catalase and decreasing the activity levels of four membrane-bound dehydrogenases. This combination would lead to lower levels of hydroxyl radicals and singlet oxygen, moieties thought to be integrally involved in the antibacterial action of HOCl. Furthermore, the resistant strain did not display the same degree of DNA damage as did the sensitive strain. CONCLUSIONS: Strain 104 is believed to grow in the presence of 28 mg l(-1) HOCl by protecting itself against HOCl by decreasing the levels of species that could react with HOCl to generate toxic reactive oxygen radicals and by improved DNA damage repair mechanisms. SIGNIFICANCE AND IMPACT OF THE STUDY: The occurrence of Salmonella able to grow in the presence of 28 mg l(-1) HOCl is of relevance to the food-processing and drinking water treatment industries as these strains would survive sanitation regimes. PMID- 11872137 TI - An overview of C. Elegans trafficking mutants. AB - It is almost 40 years since Sydney Brenner introduced Caenorhabditis elegans as a model genetic system. During that time mutants with defects in intracellular trafficking have been identified in a diverse range of screens for abnormalities. This should, of course, come as no surprise as it is hard to imagine any biological process in which the regulated movement of vesicles within the cells is not critical at some step. Almost all of these genes have mammalian homologs, and yet the role of many of these homologs has not been investigated. Perhaps the protein that regulates your favorite trafficking step has already been identified in C. elegans? Here I provide a brief overview of those trafficking mutants identified in C. elegans and where you can read more about them. PMID- 11872135 TI - Thermophilic Campylobacter spp. in turkey samples: evaluation of two automated enzyme immunoassays and conventional microbiological techniques. AB - AIMS: To determine the sensitivity and specificity of two automated enzyme immunoassays (EIA), EiaFoss and Minividas, and a conventional microbiological culture technique for detecting thermophilic Campylobacter spp. in turkey samples. METHODS AND RESULTS: A total of 286 samples (faecal, meat, neckskin and environmental samples) were collected over a period of 4 months at a turkey slaughterhouse and meat-cutting plant in Denmark. Faecal and environmental samples were tested by the conventional culture method and by the two EIAs, whereas meat and neckskin samples were tested by the two EIAs only. Two enrichment broths were used, Campylobacter Enrichment Broth (CEB) and Preston Broth (PB). Verification of positive test results was carried out by conventional culture on selective solid media. The specificities of all methods were high. The sensitivities of the EIAs were higher than that of the conventional culture technique but varied depending on the type of sample and enrichment broth. For neckskin samples, the Minividas had a significantly higher sensitivity than the EiaFoss and using PB instead of CEB as the enrichment broth significantly improved the sensitivity for both EIAs. CONCLUSIONS: Both EIAs provided more accurate results than the conventional culture technique. Furthermore, neckskin samples enriched in PB resulted in more positive test results and Campylobacter growth than samples enriched in CEB. SIGNIFICANCE AND IMPACT OF THE STUDY: The Eiafoss and Minividas proved to be reliable methods for detecting Campylobacter spp. in various samples. However, the results emphasize the need for the development of specific enrichment protocols for specific samples. PMID- 11872138 TI - Deciphering endocytosis in Caenorhabditis elegans. AB - We discuss in this review recent studies using the worm Caenorhabditis elegans to decipher endocytic trafficking in a multicellular organism. Recent advances, including in vivo assay systems, new genetic screens, comparative functional analysis of conserved proteins, and RNA-mediated interference (RNAi) in C. elegans, are being used to study the functions of known membrane trafficking factors and to identify new ones. The ability to monitor endocytosis in vivo in worms allows one to test current endocytosis models and to demonstrate the physiological significance of factors identified by genetic and biochemical methods. The available human genome sequence facilitates comparative studies where human homologs of new factors identified in C. elegans can be quickly assayed for similar function using traditional cell biological methods in mammalian cell systems. New studies in C. elegans have used a combination of these techniques to reveal novel metazoan-specific trafficking factors required for endocytosis. Many more metazoan-specific trafficking factors and insights into the mechanisms of endocytosis are likely to be uncovered by analysis in C. elegans. PMID- 11872139 TI - Metazoan motor models: kinesin superfamily in C. elegans. AB - In eukaryotic cells members of the kinesin family mediate intracellular transport by carrying cellular cargo on microtubule tracks. The nematode Caenorhabditis elegans genome encodes 21 members of the kinesin family, which show significant homology to their mammalian orthologs. Based on motor domain sequence homology and placement of the motor domain in the protein, the C. elegans kinesins have been placed in eight distinct groups; members of which participate in embryonic development, protein transport, synaptic membrane vesicles movement and in the axonal growth. Among 21 kinesins, at least 11 play a central role in spindle movement and chromosomal segregation. Understanding the function of C. elegans kinesins and related proteins may help navigate through the intricacies of intracellular traffic in a simple animal. PMID- 11872140 TI - To 5D and beyond: quantitative fluorescence microscopy in the postgenomic era. AB - Digital fluorescence microscopy is now a standard technology for assaying molecular localisation in cells and tissues. The choice of laser scanning (LSM) and wide-field microscopes (WFM) largely depends on the type of sample, with LSMs performing best on thick samples and WFMs performing best on thin ones. These systems are increasingly used to collect large multidimensional datasets. We propose a unified image structure that considers space, time, and fluorescence wavelength as integral parts of the image. Moreover, the application of fluorescence imaging to large-scale screening means that large datasets are now routinely acquired. We propose that analysis of these data requires querying tools based on relational databases and describe one such system. PMID- 11872141 TI - Ordering of compartments in the yeast endocytic pathway. AB - We have characterized the morphology of the yeast endocytic pathway leading from the plasma membrane to the vacuole by following the trafficking of positively charged nanogold in combination with compartment identification using immunolocalization of t-SNARE proteins. The first endocytic compartment, termed the early/recycling endosome, contains the t-SNARE, Tlg1p. The next compartment, the prevacuolar compartment, contains Pep12p. After transport to the prevacuolar compartment, where vacuolar enzymes are seen on their way to the vacuole, endocytic content is delivered to the late endosome and on to the vacuole, both of which are devoid of Pep12p immunolabel. Traffic to the prevacuolar compartment is reduced in strains mutant for the Rab5 homologs, Vps21p, Ypt52p, and Ypt53p and in vps27 mutant cells. On the other hand, traffic to the early recycling endosome is less dependent on Rab5 homologs and does not require Vps27p. PMID- 11872142 TI - The Dictyostelium LvsA protein is localized on the contractile vacuole and is required for osmoregulation. AB - LvsA is a Dictyostelium protein that is essential for cytokinesis and that is related to the mammalian beige/LYST family of proteins. To better understand the function of this novel protein family we tagged LvsA with GFP using recombination techniques. GFP-LvsA is primarily associated with the membranes of the contractile vacuole system and it also has a punctate distribution in the cytoplasm. Two markers of the Dictyostelium contractile vacuole, the vacuolar proton pump and calmodulin, show extensive colocalization with GFP-LvsA on contractile vacuole membranes. Interestingly, the association of LvsA with contractile vacuole membranes occurs only during the discharge phase of the vacuole. In LvsA mutants the contractile vacuole becomes disorganized and calmodulin dissociates from the contractile vacuole membranes. Consequently, the contractile vacuole is unable to function normally, it can swell but seems unable to discharge and the LvsA mutants become osmosensitive. These results demonstrate that LvsA can associate transiently with the contractile vacuole membrane compartment and that this association is necessary for the function of the contractile vacuole during osmoregulation. This transient association with specific membrane compartments may be a general property of other BEACH-domain containing proteins. PMID- 11872143 TI - Objective evaluation of differences in protein subcellular distribution. AB - Research on the cell and molecular biology of proteins involved in membrane traffic often requires determination of the effects of various experimental conditions on the subcellular distributions of those proteins. This is most often accomplished by acquiring fluorescence microscope images and visually comparing these images. While this approach is quite suitable for detecting major changes in distributions, it is not sensitive to small changes and does not permit a quantitative and objective analysis. We therefore describe the application of pattern analysis methods to the comparison of sets of fluorescence microscope images. This approach provides a high throughput and reproducible technique to determine whether image distributions differ within a specified statistical confidence, and is shown to resolve image sets indistinguishable by visual inspection. PMID- 11872145 TI - Calcium, viruses and nuclear pores: meeting report from the EMBO workshop 'Signal Transduction-Mediated Regulation of Nuclear Transport', Strasbourg, France, 11- 14 August 2001. PMID- 11872144 TI - Automated recognition of intracellular organelles in confocal microscope images. AB - Recognition of the localisation of intracellular proteins is essential to the understanding of their function. It is usually made through knowledge of and comparison to the distribution of well-characterised intracellular organelles by experts in cell biology. We have automated this process in order to achieve a more objective and quantitative assessment of the protein distribution within the cell, which can be employed by the less experienced cell biologist and may be utilised as a training program for inexperienced users, or as a high throughput localisation program for novel genes in functional analysis. Here we describe the development and testing of a classification system based on a modular neural network trained with sets of confocal sections through cell lines fluorescently stained for markers of key intracellular structures. The system functioned well in spite of the variability in pattern that occurs between individual cells and performed with 97% accuracy, which gives us confidence in the method and in its future development. It is envisaged that this program will aid the design of further experiments utilising colocalisation with known organelle marker proteins, in order to confirm putative trafficking pathways and protein--protein interactions of the protein of interest. PMID- 11872146 TI - Days and knights discussing membrane dynamics in endocytosis: meeting report from the Euresco/EMBL Membrane Dynamics in Endocytosis, 6--11 October in Tomar, Portugal. AB - The Euresco/EMBL sponsored meeting on 'Membrane Dynamics in Endocytosis' took place on 6--11 October in Tomar, Portugal. Here we report on the 5 full days of exciting talks and active poster sessions that covered topics ranging from the mechanisms of clathrin-mediated endocytosis, the regulation of phagocytosis, caveolae dynamics and function, the role of lipids in regulating endocytic transport, the formation of and sorting into and out of multivesicular bodies, new links between the actin cytoskeleton and vesicular transport, and emerging roles for endocytic trafficking in signal transduction and development. PMID- 11872147 TI - The role of serine proteases and serine protease inhibitors in the migration of gonadotropin-releasing hormone neurons. AB - BACKGROUND: Mechanisms regulating neuronal migration during development remain largely undefined. Extracellular matrix cues, target site released factors, and components of the migratory neurons themselves are likely all coordinated in time and space directing neurons to their appropriate locations. We have studied the effects of proteases and their inhibitors on the extracellular matrix and the consequences to the migration of gonadotropin releasing hormone (GnRH) neurons in the embryonic chick. Chick GnRH neurons differentiate in the olfactory epithelium, migrate along the olfactory nerve and enter the forebrain. The accessibility of this coherent cell group make it amenable for studying protease/inhibitor roles in migratory processes. RESULTS: Affigel blue beads were used to deliver a serine protease inhibitor, protease nexin-1 (PN-1), and a target protease, trypsin, to the olfactory epithelium coincident with initiation of GnRH neuronal migration. PN-1 inhibited neuronal migration while trypsin accelerated their transit into the CNS. Prior to initiation of migration, neither PN-1 nor trypsin altered the timing of neuronal exit. Trypsin did, however, accelerate the timing of neuronal crossing into the nerve-forebrain junction. CONCLUSIONS: These data support the hypothesis that protease activity modulates neuronal movements across barriers. Moreover, the data suggest, for the first time, that aspects of GnRH neuronal migration may be cell autonomous but modulated by ECM alterations. PMID- 11872148 TI - Low molecular mass dinitrosyl nonheme-iron complexes up-regulate noradrenaline release in the rat tail artery. AB - BACKGROUND: Dinitrosyl nonheme-iron complexes can appear in cells and tissues overproducing nitric oxide. It is believed that due to their chemical nature these species may be implicated in certain pathophysiological events. We studied the possible role of low molecular mass dinitrosyl iron complexes in the control of noradrenaline release in electrically stimulated rat tail artery. RESULTS: A model complex, dinitrosyl-iron-thiosulfate (at 1-10 microM) produced a concentration-dependent enhancement of electrical field stimulated [3H]noradrenaline release (up to 2 fold). At the same time, dinitrosyl-iron thiosulfate inhibited neurogenic vasoconstriction, consistent with its nitric oxide donor properties. A specific inhibitor of cyclic GMP dependent protein kinase, Rp-8pCPT-cGMPS, partially inhibited the effect of dinitrosyl-iron thiosulfate on neurogenic vasoconstriction, but not on [3H]noradrenaline release. Another model complex, dinitrosyl-iron-cysteine (at 3 microM) elicited similar responses as dinitrosyl-iron-thiosulfate. Conventional NO and NO+ donors such as sodium nitroprusside, S-nitroso-L-cysteine or S-nitroso-glutathione (at 10 microM) had no effect on [3H]noradrenaline release, though they potently decreased electrically-induced vasoconstriction. The "false complex", iron(II) thiosulfate showed no activity. CONCLUSIONS: Low molecular mass iron dinitrosyl complexes can up-regulate the stimulation-evoked release of vascular [3H]noradrenaline, apparently independently of their NO donor properties. This finding may have important implications in inflammatory tissues. PMID- 11872150 TI - A comparative study of gallstones from children and adults using FTIR spectroscopy and fluorescence microscopy. AB - BACKGROUND: Cholelithiasis is the gallstone disease (GSD) where stones are formed in the gallbladder. The main function of the gallbladder is to concentrate bile by the absorption of water and sodium. GSD has high prevalence among elderly adults. There are three major types of gallstones found in patients, White, Black and Brown. The major chemical component of white stones is cholesterol. Black and brown stones contain different proportions of cholesterol and bilirubin. The pathogenesis of gallstones is not clearly understood. Analysis of the chemical composition of gallstones using various spectroscopic techniques offers clues to the pathogenesis of gallstones. Recent years has seen an increasing trend in the number of cases involving children. The focus of this study is on the analysis of the chemical composition of gallstones from child and adult patients using spectroscopic methods. METHODS: In this report, we present FTIR spectroscopic studies and fluorescence microscopic analysis of gallstones obtained from 67 adult and 21 child patients. The gallstones were removed during surgical operations at Soroka University Medical Center. RESULTS: Our results show that black stones from adults and children are rich in bilirubin. Brown stones are composed of varying amounts of bilirubin and cholesterol. Green stones removed from an adult, which is rare, was found to be composed mainly of cholesterol. Our results also indicated that cholesterol and bilirubin could be the risk factors for gallstone formation in adults and children respectively. Fluorescence micrographs showed that the Ca-bilirubinate was present in all stones in different quantities and however, Cu-bilirubinate was present only in the mixed and black stones. CONCLUSIONS: Analysis based on FTIR suggest that the composition of black and brown stones from both children and adults are similar. Various layers of the brown stone from adults differ by having varying quantities of cholesterol and calcium carbonate. Ring patterns observed mainly in the green stone using fluorescence microscopy have relevance to the mechanism of the stone formation. Our preliminary study suggests that bilirubin and cholesterol are the main risk factors of gallstone disease. PMID- 11872149 TI - 4-(N,N-dipropylamino)benzaldehyde inhibits the oxidation of all-trans retinal to all-trans retinoic acid by ALDH1A1, but not the differentiation of HL-60 promyelocytic leukemia cells exposed to all-trans retinal. AB - BACKGROUND: The signal transduction pathways mediated by retinoic acid play a critical role in the regulation of cell growth and differentiation during embryogenesis and hematopoiesis as well as in a variety of tumor cell lines in culture. Following the reports that two members of the superfamily of aldehyde dehydrogenase (ALDH) enzymes, ALDH1A1 and ALDH1A2, were capable of catalyzing the oxidation of all-trans retinal to all-trans retinoic acid with submicromolar Km values, we initiated an investigation of the ability of 4-(N,N dipropylamino)benzaldehyde (DPAB) to inhibit the oxidation of retinal by purified mouse and human ALDH1A1. RESULTS: Our results show that DPAB potently inhibits retinal oxidation, with IC50 values of 0.11 and 0.13 microM for purified mouse and human ALDH1A1, respectively. Since the HL-60 human myeloid leukemic cell line has been used extensively to study the retinoic acid induced differentiation of HL-60 cells to granulocytes, and ALDH1A1 activity had previously been reported in HL-60 cells, we investigated the ability of DPAB to block differentiation of HL 60 promyelocytic leukemia cells exposed to retinal in culture. In HL-60 cells coincubated with 1 microM retinal and 50 microM DPAB for 144 hours, cell differentiation was inhibited only 30%. Furthermore, the NAD-dependent oxidation of propanal or retinal was less than 0.05 nmoles NADH formed/min-10(7) cells in spectrophotometric assays using HL-60 cell extracts. CONCLUSION: Although ALDH1A1 may be the major catalytic activity for retinal oxidation in some retinoid dependent mouse and Xenopus embryonic tissues and in adult human and mouse hematopoietic stem cells, another catalytic activity appears to synthesize the retinoic acid ligand necessary to stimulate the differentiation of HL-60 cells to end stage granulocytes. PMID- 11872153 TI - A new chemical formulation for control of dental unit water line contamination: An 'in vitro' and clinical 'study' AB - BACKGROUND: Water delivered by dental units during routine dental practice is highly contaminated. The aim of this study is to evaluate the efficacy of a new chemical solution flushed through Dental Unit Water Lines (DUWL) for the control of contamination inside dental units. MATERIALS AND METHODS: Six old dental units equipped with a device designed to automatically flush disinfecting solutions through the water system (Castellini Autosteril) were selected. Water samples from DUWL effluents were collected in each dental unit for 10 randomly selected days, before and after a 5 minute DUWL disinfecting cycle with TetraAcetylEthileneDiamine (TAED) and persalt (Ster4spray produced by Farmec spa, and distributed by Castellini spa). Water samples were plated in R2A Agar and cultured at room temperature for 7 days, and the total number of heterotrophic microorganisms counted and expressed in Log10 CFU/mL A general linear model was fitted and multiple regression ANOVA for repeated measures was used for the statistical analysis. RESULTS: The mean contamination in DUWL effluent at baseline was 5.45 &#PlusMinus; 0.35 CFU/mL (range 4.79 to 5.93 CFU/mL). When water samples were tested "in vitro" against the chemical, no growth of heterotrophic bacteria was detected after a 5 minute contact in any of the water samples tested. After undergoing a 5 minute disinfecting cycle with the chemical, DUWL mean contamination in water effluents was 2.01 &#PlusMinus; 0.32 CFU/mL (range 1.30 to 2.74 CFU/mL) (significant difference with respect to baseline). CONCLUSIONS: An inbetween patient disinfecting procedure consisting of flushing DUWL with TAED and persalt equivalent to 0.26% peracetic acid could be useful in routine dental practice for cross-contamination control. PMID- 11872152 TI - A role for the collagen I/III and MMP-1/-13 genes in primary inguinal hernia? AB - BACKGROUND: Abnormal collagen metabolism is thought to play an important role in the development of primary inguinal hernia. This is underlined by detection of altered collagen metabolism and structural changes of the tissue in patients with primary inguinal hernia. However, it is still unknown whether these alterations reflect a basic dysfunction of the collagen synthesis, or of collagen degradation. METHODS: In the present study, we analysed type I and type III procollagen messenger ribonucleic acid (mRNA) and MMP-1 and MMP-13 mRNA in cultured fibroblasts from the skin of patients with primary inguinal hernia, and from patients without hernia (controls) by reverse transcription polymerase chain reaction (RT-PCR) and Northern Blot. RESULTS: The results indicated that the ratio of type I to type III procollagen mRNA was decreased in patients with primary hernia, showing significant differences as compared to controls (p = 0.01). This decrease was mainly due to the increase of type III procollagen mRNA. Furthermore, RT-PCR analysis revealed that the expression of MMP-1 mRNA in patients with primary hernia is equivalent to that of controls (p > 0.05). In addition, MMP-13 mRNA is expressed neither in patients with primary hernia nor in controls. CONCLUSION: We concluded that abnormal change of type I and type III collagen mRNAs contribute to the development of primary inguinal hernia, whereas the expressions of MMP-1 and MMP-13 mRNA appears not to be involved in the development of primary inguinal hernia. Thus, the knowledge on the transcriptional regulation of collagen in patients with primary inguinal hernia may help to understand the pathogenesis of primary inguinal hernia, and implies new therapeutic strategies for this disease. PMID- 11872154 TI - A thematic series on lipid signaling: prologue. PMID- 11872151 TI - Evidence for modulation of pericryptal sheath myofibroblasts in rat descending colon by transforming growth factor beta and angiotensin II. AB - BACKGROUND: Absorption of water and Na+ in descending colonic crypts is dependent on the barrier function of the surrounding myofibroblastic pericryptal sheath. Here the effects of high and low Na+ diets and exposure to whole body ionising radiation on the growth and activation of the descending colonic pericryptal myofibroblasts are evaluated. In addition the effect of a post-irradiation treatment with the angiotensin converting enzyme inhibitor Captopril was investigated. METHODS: The levels of Angiotensin II type 1 receptor (AT1), ACE, collagen type IV, transforming growth factor-beta type 1 receptor (TGF-betaR1), OB cadherin and alpha-smooth muscle actin in both descending colon and caecum were evaluated, using immunocytochemistry and confocal microscopy, in rats fed on high and low Na+ diets (LS). These parameters were also determined during 3 months post-irradiation with 8Gy from a 60Co source in the presence and absence of the angiotensin converting enzyme inhibitor, Captopril. RESULTS: Increases in AT1 receptor (135.6% +/- 18.3, P < 0.001); ACE (70.1% +/- 13.1, P < 0.001); collagen type IV (49.6% +/- 15.3, P < 0.001); TGF-+/-beta1 receptors (291.0% +/- 26.5, P < 0.001); OB-cadherin (26.3% +/- 13.8, P < 0.05) and alpha-smooth muscle actin (82.5% +/- 12.4, P < 0.001) were observed in the pericryptal myofibroblasts of the descending colon after LS diet. There are also increases in AT1 receptor and TGF-beta1 receptor, smooth muscle actin and collagen type IV after irradiation. Captopril reduced all these effects of irradiation on the pericryptal sheath and also decreased the amount of collagen and smooth muscle actin in control rats (P < 0.001). CONCLUSIONS: These results demonstrate an activation of descending colonic myofibroblasts to trophic stimuli, or irradiation, which can be attenuated by Captopril, indicative of local trophic control by angiotensin II and TGF-beta release. PMID- 11872155 TI - Phospholipase A2. AB - Phospholipase A2 (PLA2) catalyzes the hydrolysis of the sn-2 position of membrane glycerophospholipids to liberate arachidonic acid (AA), a precursor of eicosanoids including prostaglandins (PGs) and leukotrienes (LTs). The same reaction also produces lysophosholipids, which represent another class of lipid mediators. So far, at least 19 enzymes that possess PLA2 activity have been identified in mammals. The secretory PLA2 (sPLA2) family, in which 10 isozymes have been identified, consists of low-molecular-weight, Ca2+-requiring, secretory enzymes that have been implicated in a number of biological processes, such as modification of eicosanoid generation, inflammation, host defense, and atherosclerosis. The cytosolic PLA2 (cPLA2) family consists of 3 enzymes, among which cPLA2alpha plays an essential role in the initiation of AA metabolism. Intracellular activation of cPLA2alpha is tightly regulated by Ca2+ and phosphorylation. The Ca2+-independent PLA2 (iPLA2) family contains 2 enzymes and may play a major role in membrane phospholipid remodeling. The platelet activating factor (PAF) acetylhydrolase (PAF-AH) family represents a unique group of PLA2 that contains 4 enzymes exhibiting unusual substrate specificity toward PAF and/or oxidized phospholipids. In this review, we will overview current understanding of the properties and functions of each enzyme belonging to the sPLA2, cPLA2, and iPLA2 families, which have been implicated in signal transduction. PMID- 11872157 TI - Regulation and possible role of mammalian phospholipase D in cellular functions. PMID- 11872156 TI - Structure, regulation, and function of phospholipase C isozymes. PMID- 11872158 TI - The second largest subunit of mouse DNA polymerase epsilon, DPE2, interacts with SAP18 and recruits the Sin3 co-repressor protein to DNA. AB - DNA polymerase epsilon is essential for cell viability and chromosomal DNA replication in budding yeast. In addition, DNA polymerase epsilon may be involved in DNA repair and cell-cycle checkpoint control. The enzyme consists of at least four subunits in mammalian cells as well as in yeast. The largest subunit of DNA polymerase epsilon is responsible for polymerase activity. To date, the functions of the other subunits have remained unknown. With a view to elucidating the functions of the second largest subunit of mouse DNA polymerase epsilon (DPE2), yeast two-hybrid screening was performed to identify mouse proteins that interact with this subunit. SAP18, a polypeptide associated with co-repressor protein Sin3, was identified as an interacting protein. A part of the N-terminal region of DPE2 (comprising amino acids 85-250) was found to be responsible for the interaction with SAP18. The interaction induced repression of transcription in reporter plasmid assays, which was inhibited by trichostatin A. These results indicate that DPE2 may recruit histone deacetylase (HDAC) to the replication fork to modify the chromatin structure. PMID- 11872159 TI - Crystal structure of 1-deoxy-D-xylulose 5-phosphate reductoisomerase complexed with cofactors: implications of a flexible loop movement upon substrate binding. AB - The key enzyme in the nonmevalonate pathway, 1-deoxy-D-xylulose 5-phosphate reductoisomerase (DXR), has been shown to be an effective target of antimalarial drugs. Here we report the crystal structure of DXR complexed with NADPH and a sulfate ion from Escherichia coli at 2.2 A resolution. The structure showed the presence of an extra domain, which is absent from other NADPH-dependent oxidoreductases, in addition to the conformation of catalytic residues and the substrate binding site. A flexible loop covering the substrate binding site plays an important role in the enzymatic reaction and the determination of substrate specificity. PMID- 11872160 TI - Proteasome inhibitors block Ras/ERK signaling pathway resulting in the downregulation of Fas ligand expression during activation-induced cell death in T cells. AB - Activation-induced cell death (AICD) plays a critical role in the maintenance of homeostasis and peripheral tolerance in the immune system, and is mediated by Fas ligand (FasL) expression and the interaction between Fas and FasL. In the present study, we examined the role of the ubiquitin-proteasome system in AICD using T cell hybridoma N3-6-71 cells. The peptidyl aldehyde proteasome inhibitor carbobenzoxyl-Ile-Glu(O-t-butyl)-Ala-leucinal (PSI) blocked T cell receptor (TCR) stimulation-induced apoptosis in the T cell hybridoma. Fas and FasL gene expression and mouse FasL promoter activity following TCR stimulation were suppressed by PSI pretreatment. Deletion or point mutation of the kappaB site in the FasL promoter region did not suppress inducible FasL promoter activity effectively. PSI blocked extracellular signal-regulated kinase (ERK) activity induced by TCR stimulation, but had no effect on c-jun N-terminal kinase activation. ERK activation was essential for FasL expression and AICD. The initial tyrosine phosphorylation steps following TCR stimulation, i.e., phosphorylation of CD3zeta and Vav, were not altered by PSI. These data suggest that the ubiquitin-proteasome system has some regulatory function at an intermediate step between the initial tyrosine phosphorylation steps and ERK activation in AICD. PMID- 11872161 TI - Gamma-adaptin interacts directly with Rabaptin-5 through its ear domain. AB - In yeast two-hybrid screening using gamma1-adaptin, a subunit of the AP-1 adaptor complex of clathrin-coated vesicles derived from the trans-Golgi network (TGN), as bait, we found that it could interact with Rabaptin-5, an effector of Rab5 and Rab4 that regulates membrane docking with endosomes. Further two-hybrid analysis revealed that the interaction occurs between the ear domain of gamma1-adaptin and the COOH-terminal coiled-coil region of Rabaptin-5. Pull down assay with a fusion protein between glutathione S-transferase and the ear domain of gamma1-adaptin and coimmunoprecipitation analysis revealed that the interaction occurs in vitro and in vivo. Immunocytochemical analysis showed that gamma1-adaptin and Rabaptin 5 colocalize to a significant extent on perinuclear structures, probably on recycling endosomes, and are redistributed into the cytoplasm upon treatment with brefeldin A. These results suggest that the gamma1-adaptin-Rabaptin-5 interaction may play a role in membrane trafficking between the TGN and endosomes. PMID- 11872162 TI - Molecular cloning and genomic analysis of mouse glucuronyltransferase involved in biosynthesis of the HNK-1 epitope. AB - cDNA and genomic clones encoding the mouse glucuronyltransferase (GlcAT-P) involved in biosynthesis of the HNK-1 carbohydrate epitope were isolated and the structural organization of the gene was determined. The predicted amino acid sequence of mouse GlcAT-P is 96.2 and 98.2% identical to those of the rat and human enzymes, respectively. Alternatively spliced isoforms of mouse GlcAT-P are present in the brain and encode two proteins that are identical throughout their length except for an additional 13 amino acids in the N-terminal cytoplasmic domain of the major form. The coding region of GlcAT-P is composed of 5 exons spanning approximately 6 kb, and the GlcAT-P gene was mapped to the A4 region of mouse chromosome 9. Upstream of the transcriptional start site, no typical TATA or CCAAT box was found, but binding sites for several known transcription factors including Sp1 and Krox-20 were identified. Transient transfection of luciferase reporter constructs demonstrated that a 207 bp fragment of the 5'-upstream region acts as a strong promoter in PC-12 cells, which express the HNK-1 epitope, but not in COS-1 cells. Thus, this minimal promoter region of GlcAT-P is suggested to be associated with the regulation of HNK-1 expression. PMID- 11872163 TI - Characterization of the biological functions of a transcription factor, c-myc intron binding protein 1 (MIBP1). AB - The c-myc intron binding protein 1 (MIBP1) is a gigantic zinc finger protein comprising 2,437 amino acids and belonging to the MHC binding protein (MBP) family. MIBP1 is suggested to be a transcription factor involved in various biological functions. We show here that MIBP1 represses c-myc transcription from the major promoter, P2. Screening by the yeast two-hybrid system revealed that the MIBP1 protein interacts with the Ski-interacting protein (SKIP). In vitro pull-down assays and in vivo co-immunoprecipitation experiments confirmed this interaction. The acidic region of MIBP1 was found to be the site of interaction with the N-terminal half of SKIP. In situ hybridization analysis using developing rat embryos revealed that the MIBP1 mRNA is highly expressed in post-mitotic neurons, but the expression in immature neuroepithelium is low. The expression of MIBP1 in adult rat brain is also predominantly in neuronal cells, indicating that MIBP1 is involved in the physiology of mature neuronal cells. PMID- 11872164 TI - Determination of the specific interaction between sulfonylurea-incorporated polymer and rat islets. AB - A SU derivative, mimicking glibenclamide in chemical structure, was synthesized to incorporate it into a water-soluble polymeric backbone as a biospecific and stimulating polymer for insulin secretion. The ability of insulin secretion was examined with different glucose concentrations (3.3 and 11.6 mM). Although the vinylated SU did not exhibit significant activity compared to the control, the SU incorporated copolymer could enhance insulin secretion as much as or more than glibenclamide did. In this study, a polymer fluorescence-labeled with rodamine-B isothiocyanate was used to visualize the interactions and we found that the labeled polymer was strongly absorbed to rat islets, probably due to its specific interaction mediated by SU receptors on the cell membrane. To verify the specific interaction between the SU (K+ channel closer)-incorporated copolymer and rat islets, cells were pretreated with diazoxide, an agonist of ATP-sensitive K+ channels (K+ channel opener), before adding the incorporated polymer to the cell culture medium. This treatment suppressed the action of SUs on rat islets. A confocal laser microscopic study further confirmed this interaction. The results of this study provided evidence that the SU-incorporated copolymer stimulates insulin secretion through specific interactions of SU moieties in the polymer with rat islets. PMID- 11872165 TI - Three-dimensional structure of the flavoenzyme acyl-CoA oxidase-II from rat liver, the peroxisomal counterpart of mitochondrial acyl-CoA dehydrogenase. AB - Acyl-CoA oxidase (ACO) catalyzes the first and rate-determining step of the peroxisomal beta-oxidation of fatty acids. The crystal structure of ACO-II, which is one of two forms of rat liver ACO (ACO-I and ACO-II), has been solved and refined to an R-factor of 20.6% at 2.2-A resolution. The enzyme is a homodimer, and the polypeptide chain of the subunit is folded into the N-terminal alpha domain, beta-domain, and C-terminal alpha-domain. The X-ray analysis showed that the overall folding of ACO-II less C-terminal 221 residues is similar to that of medium-chain acyl-CoA dehydrogenase (MCAD). However, the N-terminal alpha- and beta-domains rotate by 13 with respect to the C-terminal alpha-domain compared with those in MCAD to give a long and large crevice that accommodates the cofactor FAD and the substrate acyl-CoA. FAD is bound to the crevice between the beta- and C-terminal domains with its adenosine diphosphate portion interacting extensively with the other subunit of the molecule. The flavin ring of FAD resides at the active site with its si-face attached to the beta-domain, and is surrounded by active-site residues in a mode similar to that found in MCAD. However, the residues have weak interactions with the flavin ring due to the loss of some of the important hydrogen bonds with the flavin ring found in MCAD. The catalytic residue Glu421 in the C-terminal alpha-domain seems to be too far away from the flavin ring to abstract the alpha-proton of the substrate acyl-CoA, suggesting that the C-terminal domain moves to close the active site upon substrate binding. The pyrimidine moiety of flavin is exposed to the solvent and can readily be attacked by molecular oxygen, while that in MCAD is protected from the solvent. The crevice for binding the fatty acyl chain is 28 A long and 6 A wide, large enough to accommodate the C23 acyl chain. PMID- 11872166 TI - Coupling of calcium transport with ATP hydrolysis in scallop sarcoplasmic reticulum. AB - Previously, we showed that incubation of the scallop sarcoplasmic reticulum (SR) with EGTA at above 37 degrees C resulted in the uncoupling of ATP hydrolysis with Ca2+ transport [Nagata et al. (1996) J. Biochem. 119, 1100-1105]. We have extended this study by comparing the kinetic behavior of Ca2+ release and binding to the uncoupled SR with that of intact scallop or rabbit SR. The change in the Ca2+ concentration in the reaction medium, as determined as the absorption of APIII, was followed using a stopped flow system. Intact scallop SR was preincubated with Ca2+ in the presence of a Ca2+ ionophore, A23187, and then ATP was added to initiate the reaction. The Ca2+ level in the medium increased to the maximum level in several seconds, and then slowly decreased to the initial low level. The rising and subsequent slow decay phases could be related to the dissociation and reassociation of Ca2+ with the Ca-ATPase, respectively. When uncoupled scallop SR vesicles were preincubated with CaCl2 in the absence of A23187 and then the reaction was initiated by the addition of ATP, a remarkable amount of Ca2+ was released from the SR vesicles into the cytosolic solution, whereas, with intact scallop or rabbit SR, only a sharp decrease in the Ca2+ level was observed. Based on these findings, we concluded that the heat treatment of scallop SR in EGTA may alter the conformation of the Ca-ATPase, thereby causing Ca2+ to be released from the enzyme, during the catalytic cycle, at the cytoplasmic surface, but not at the lumenal surface of SR vesicles. PMID- 11872167 TI - Expression of NADP+-dependent 15-hydroxyprostaglandin dehydrogenase mRNA in monkey ocular tissues and characterization of its recombinant enzyme. AB - Although prostaglandin (PG) F2a and its analogue latanoprost decrease the intraocular pressure in a variety of animals, their intraocular metabolism has not yet been clarified. Here, we isolated the cDNAs for the monkey homologues of NAD+- and NADP+-dependent types of 15-hydroxy PG dehydrogenase (PGDH) from lung and eye, respectively, and investigated the distribution of their mRNAs in the monkey eye. The cDNAs for the NAD+- and NADP+-dependent types of PGDH contained an open reading frame of 798 and 831 bp encoding 266 and 277 amino acid residues with calculated molecular masses of 28.9 and 30.5 kDa, respectively. The amino acid sequences of the monkey NAD+- and NADP+-dependent enzymes showed less than 20% identity to each other, and the former enzyme shows 98.5 and 86.8% identity, and the latter 94.9 and 85.2% identity, to the human and mouse enzymes, respectively. Reverse transcription-PCR analysis revealed that the mRNA for NADP+ dependent PGDH, but not that for NAD+-dependent PGDH, was highly expressed in monkey ocular tissues. In situ hybridization analysis demonstrated that the mRNA for NADP+-dependent PGDH was localized in the epithelial cells of the cornea. The recombinant NADP+-dependent PGDH catalyzed the dehydrogenation of the 15-hydroxyl group of PGF2a and the acid form of latanoprost in the presence of NADP+ as examined by HPLC. These results indicate that PGF2a and the acid form of latanoprost are degraded to their 15-keto metabolites by NADP+-dependent PGDH localized in the monkey eye. PMID- 11872168 TI - Genes for a nuclease and a protease are involved in the drastic decrease in cellular RNA amount in fission yeast cells during nitrogen starvation. AB - Cellular RNA in Schizosaccharomyces pombe cells drastically decreases in amount during nitrogen starvation. Previously, we found and purified a soluble RNA degrading enzyme whose activity drastically increased in the cells of S. pombe undergoing nitrogen starvation. The enzyme was a nuclease encoded by pnu1(+). In this study, the increase in the RNA-degrading activity and the decrease in cellular RNA level are examined in a null-mutant of pnu1(+) (pnu1Delta). During nitrogen starvation, wild-type cells show an apparent increase in RNA-degrading activity, whereas the pnu1Delta cells do not. The wild-type cells show a drastic decrease in cellular RNA amount, whereas the pnu1Delta cells show only a slight decrease. These results suggest that Pnu1 nuclease is implicated in the decrease in cellular RNA amount during nitrogen starvation, probably via the RNA-degrading activity. The increase in the RNA-degrading activity is independent of both the Wis1 stress-activated MAP kinase cascade and Tor1 signaling pathway, but it is strongly dependent on isp6(+), a gene for a possible protease, whose expression is induced during nitrogen starvation. A disruption mutant for isp6(+) (isp6Delta) is deficient in both the increase in the RNA-degrading activity and the drastic decrease in the cellular RNA amount during nitrogen starvation, which suggests that isp6(+) is involved in the RNA degradation via regulating the RNA degrading activity of Pnu1. PMID- 11872169 TI - Employment of the human estrogen receptor beta ligand-binding domain and co activator SRC1 nuclear receptor-binding domain for the construction of a yeast two-hybrid detection system for endocrine disrupters. AB - To screen a wide variety of chemicals for endocrine disrupters, and to develop an effective microbial degradation system for them, a good system is needed for the rapid and accurate evaluation of the endocrine-disrupting activities of suspected chemicals and their degradation products. We constructed two-hybrid systems that co-express the Gal4p DNA binding domain/ligand-binding domain of human estrogen receptor (hER) alpha or beta and the Gal4p transactivation domain/nuclear receptor-binding domain of co-activator SRC1, TIF2, or AIB1 in Saccharomyces cerevisiae with a chromosome-integrated lacZ reporter gene under the control of Gal4p-binding sites. We found that the combination of the hERbeta ligand-binding domain and SRC1 nuclear receptor-binding domain was most effective for the xenoestrogen-dependent induction of reporter activity. The extent of transcriptional activation by known xenoestrogens and phytoestrogens was found to correlate well with their estrogenic activities as measured by the previous system with rat ERalpha. This system detects estrogenic activity in some chemicals that have not been suspected of being positive. We also applied this assay system to test the microbial degradation products of gamma hexachlorocyclohexane (gamma-HCH) by Sphingomonas paucimobilis. Among the gamma HCH metabolites, 2,5-dichlorohydroquinone and chlorohydroquinone had estrogenic activities similar to the original chemical, while hydroquinone, a later stage metabolite, showed no activity, suggesting the necessity of evaluating intermediate metabolites in microbial degradation systems. PMID- 11872170 TI - Ca2+- and S1-induced conformational changes of reconstituted skeletal muscle thin filaments observed by fluorescence energy transfer spectroscopy: structural evidence for three States of thin filament. AB - Rabbit skeletal muscle alpha-tropomyosin (Tm) and the deletion mutant (D234Tm) in which internal actin-binding pseudo-repeats 2, 3, and 4 are missing [Landis et al. (1997) J. Biol. Chem. 272, 14051-14056] were used to investigate the interaction between actin and tropomyosin or actin and troponin (Tn) by means of fluorescence resonance energy transfer (FRET). FRET between Cys-190 of D234Tm and Gln-41 or Cys-374 of actin did not cause any significant Ca2+-induced movement of D234Tm, as reported previously for native Tm [Miki et al. (1998) J. Biochem. 123, 1104-1111]. FRET did not show any significant S1-induced movement of Tm and D234Tm on thin filaments either. The distances between Cys-133 of TnI, and Gln-41 and Cys-374 of actin on thin filaments reconstituted with D234Tm (mutant thin filaments) were almost the same as those on thin filaments with native Tm (wild type thin filaments) in the absence of Ca2+. Upon binding of Ca2+ to TnC, these distances on mutant thin filaments increased by approximately 10 A in the same way as on wild-type thin filaments, which corresponds to a Ca2+-induced conformational change of thin filaments [Miki et al. (1998) J. Biochem. 123, 324 331]. The rigor binding of myosin subfragment 1 (S1) further increased these distances by approximately 7 A on both wild-type and mutant thin filaments when the thin filaments were fully decorated with S1. This indicates that a further conformational change on thin filaments was induced by S1 rigor-binding (S1 induced or open state). Plots of the extent of S1-induced conformational change vs. molar ratio of S1 to actin showed that the curve for wild-type thin filaments is hyperbolic, whereas that for mutant thin filaments is sigmoidal. This suggests that the transition to the S1-induced state on mutant thin filaments is depressed with a low population of rigor S1. In the absence of Ca2+, the distance also increased on both wild-type and mutant thin filaments close to the level in the presence of Ca2+ as the molar ratio of S1 to actin increased up to 1. The curves are sigmoidal for both wild-type and mutant thin filaments. The addition of ATP completely reversed the changes in FRET induced by rigor S1 binding. For mutant thin filaments, the transition from the closed state to the open state in the presence of ATP is strongly depressed, which results in the inhibition of acto myosin ATPase even in the presence of Ca2+. The present FRET measurements provide structural evidence for three states of thin filaments (relaxed, Ca2+-induced or closed, and S1-induced or open states) for the regulation mechanism of skeletal muscle contraction. PMID- 11872171 TI - Sequence-independent DNA binding activity of DnaA protein, the initiator of chromosomal DNA replication in Escherichia coli. AB - The DnaA protein specifically binds to the origin of chromosomal DNA replication and initiates DNA synthesis. In addition to this sequence-specific DNA binding, DnaA protein binds to DNA in a sequence-independent manner. We here compared the two DNA binding activities. Binding of ATP and ADP to DnaA inhibited the sequence independent DNA binding, but not sequence-specific binding. Sequence-independent DNA binding, but not sequence-specific binding, required incubation at high temperatures. Mutations in the C-terminal domain affected the sequence independent DNA binding activity less drastically than they did the sequence specific binding. On the other hand, the mutant DnaA433, which has mutations in a membrane-binding domain (K327 to I344) was inert for sequence-independent binding, but could bind specifically to DNA. These results suggest that the two DNA binding activities involve different domains and perform different functions from each other in Escherichia coli cells. PMID- 11872172 TI - Second derivative fluorescence spectra of indole compounds. AB - The fluorescence emission spectrum of N-acetyl tryptophan amide (NATA) in 20 mM K phosphate buffer, pH 7.5, with excitation at 295 nm, when subjected to second derivatization, showed two troughs at 340 1.0 nm (A) and 358.5 1.0 nm (B). Linear dependence of derivative intensities at A and B was observed with increasing NATA concentration between 0-30 nM but the intensity ratio (B/A), termed R, was found to be invariant at 0.70 0.05. R remained unaffected with variation of the pH (4 10), temperature (15-70 degrees C), salt concentration (0-2 M NaCl), and excitation wavelength between 280-300 nm. A 50-fold molar excess of N-acetyl tyrosine over 10 nM NATA and inclusion of a quencher like 0.8 M acrylamide, 0.4 M potassium iodide or trichloroethanol had no effect on R. It was, however, linearly dependent on the polarity of the solvent-in 1,4-dioxane it became 0.07 0.05. Derivative spectra of tryptophans of proteins largely resembled that of NATA. Low R values of between 0.02-0.34 were observed for proteins under native conditions, which is consistent with the general buried character of tryptophan residues. R increased to 0.6-0.9 after unfolding with denaturants or extensive proteolysis and decreased to close to the original value after refolding. The equilibrium unfolding transitions of proteins expressed as R largely resembled the transitions measured using other physical parameters. R appears to be a more sensitive index for monitoring the hydrophobic environment of tryptophans in protein compared to parameters like emission maxima or intensity of underivatised spectra. PMID- 11872173 TI - Effect of antifungal azoles on the heme detoxification system of malarial parasite. AB - The antimalarial activities of some antifungal azole agents (ketoconazole, miconazole, and clotrimazole) have been known for several years, however, their antimalarial mechanism remains equivocal. Our recent study showed that clotrimazole has a relative high affinity for heme, inhibits reduced glutathione dependent heme catabolism, and enhances heme-induced hemolysis. In the present study, we have found that clotrimazole can remove heme from histidine rich peptide-heme complex, which initiates heme-polymerization in malaria. In addition, we show that two other azoles (ketoconazole and miconazole) behave similarly to clotrimazole in binding to heme: they bind to heme with similar affinities, remove heme from the histidine rich peptide-heme complex and from the reduced glutathione-heme complex to form stable heme-azole complexes with two nitrogenous ligands derived from the imidazole moieties of two azole molecules. We have also revealed that clotrimazole and miconazole have stronger promoting activities for heme-induced hemolysis than ketoconazole, implying that the stronger antimalarial activities of clotrimazole and miconazole might arise from their stronger ability to promote heme-induced hemolysis of clotrimazole and clotrimazole than that of ketoconazole. These results also suggest that ketoconazole and miconazole, like clotrimazole, might possess an antimalarial mechanism relating to their inhibition of heme polymerization and the degradation of reduced glutathione-dependent heme. PMID- 11872175 TI - Presence and features of fatty acyl-CoA binding activity in rat hepatic peroxisomes. AB - We studied the fatty acyl-CoA binding activity of rat liver peroxisomes. After subcellular fractionation of rat liver treated with or without clofibrate, a peroxisome proliferator, the binding activity with [1-(14)C]palmitoyl-CoA was detected in the light mitochondrial fraction in addition to the mitochondrial and cytosol fractions. After Nycodenz centrifugation of the light mitochondrial fraction, the binding activity was detected in peroxisomes. The peroxisomal activity depended on the incubation temperature and peroxisome concentration. The activity also depended on the concentration of 2-mercaptoethanol, and a plateau of activity was unexpectedly found at 2-mercaptoethanol concentrations from 20 to 40 mM. Clofibrate increased the total and specific activity of the fatty acyl-CoA binding of peroxisomes by 7.9 and 2.5 times compared with the control, respectively. In the presence of 20% glycerol at 0 degree C, approximately 90% of the binding activity was maintained for up to at least 3 wk. After successive treatment with an ultramembrane Amicon YM series, about 70% of the binding activity was detected in the M.W. 30,000-100,000 fraction. When the M.W. 30,000 100,000 fraction was added to the incubation mixture of the peroxisomal fatty acyl-CoA beta-oxidation system, a slight increase in the beta-oxidation activity was found. 2-Mercaptoethanol (20 mM) significantly activated the fatty acyl-CoA beta-oxidation system to 1.4 times control. After gel filtration of the M.W. 30,000-100,000 fraction, the peaks of fatty acyl-CoA binding protein showed broad elution profiles from 45,000 to 75,000. These results suggest that fatty acyl-CoA binding activity can be detected directly in peroxisomes and is increased by peroxisome proliferators. The high binding activity in the presence of higher concentrations of 2-mercaptoethanol indicates the importance of the SH group for binding. The apparent molecular weight of the binding protein may be from 45,000 to 75,000. PMID- 11872174 TI - A cytosolic form of aminopeptidase P from Drosophila melanogaster: molecular cloning and characterization. AB - Using a functional genomic approach, we have identified and characterized a cytosolic form of aminopeptidase P from Drosophila melanogaster. This study represents the first characterization of an insect aminopeptidase P. The complete sequence of a 12.5 kbp genomic clone from D. melanogaster showed the presence of a 1,839 bp ORF, encoding a protein of 613 amino acids with a calculated molecular mass of 68.5 kDa. The deduced amino acid sequence was 48% identical and 66% similar to rat and human cytosolic aminopeptidase P. Amino acids important for catalytic activity and the metal binding ligands were found to be conserved between Drosophila AP-P and its mammalian homologues. The recombinant enzyme expressed in Escherichia coli hydrolyzed the amino terminal Xaa-Pro bond of substance P and bradykinin, revealing its functional identity. Further enzyme characterization showed the enzyme to be a manganese-dependent metallopeptidase. Immunoblot analysis showed that DAP-P is located exclusively in the cytosol and is temporally regulated during Drosophila development. In the adult fly, the protein could be detected in gut, testis and ovary, with a high level of expression in brain. PMID- 11872176 TI - Close location of the first loop to the third loop of the mitochondrial ADP/ATP carrier deduced from cross-linking catalyzed by copper-o-phenanthroline of the solubilized carrier with Triton X-100. AB - Effects of the cross-linking catalyst copper-o-phenanthroline [Cu(OP)2] on the bovine heart mitochondrial ADP/ATP carrier solubilized with Triton X-100 were studied under various conditions. Without detergent treatment, Cu(OP)2 specifically catalyzed the formation of intermolecular disulfide bridges in submitochondrial particles between two Cys56 residues in the first loop facing the matrix space of the dimeric carrier [Majima, E., Ikawa, K., Takeda, M., Hashimoto, M., Shinohara, Y., and Terada, H. (1995) J. Biol. Chem. 270, 29548 29554]. However, an intramolecular disulfide bridge between Cys56 and Cys256 in the third loop was formed in the solubilized carrier. Proteolytic digestion of the carrier with lysylendopeptidase showed that it first cleaves the Lys42-Gln43 bond and then the Lys48-Gln49 bond of the first loop in the membrane-bound carrier, but it cleaves both sites almost simultaneously in the solubilized carrier. These features were observed only with the m-state carrier; the c-state carrier was not subject to any cross-linking or proteolytic digestion. It is suggested that the protruding first loop is located close to the third loop, which could be exposed to a certain degree. PMID- 11872177 TI - Activation of caspase-3, proteolytic cleavage of DFF and no oligonucleosomal DNA fragmentation in apoptotic Molt-4 cells. AB - A variety of endonucleases has been implicated in apoptotic DNA fragmentation. DNA fragmentation factor (DFF) is one of the endonucleases responsible for DNA fragmentation. Since an oligonucleosomal DNA ladder is not induced in apoptotic Molt-4 cells, we investigated whether or not the absence of ladder formation is related to an inability of DFF endonuclease in the cells. Semiquantitative RT-PCR analysis showed that the mRNA level of DFF-40 and DFF-45 in Molt-4 cells was approximately the same, compared with in other cells, which exhibit different levels of the fragmentation in apoptosis. When Molt-4 cells were induced to undergo apoptosis by neocarzinostatin (NCS) treatment, both caspase-3 activation and DFF-45 cleavage were observed. Furthermore, DFF immunoprecipitated from Molt 4 cells exhibited DNA degradation activity. These results suggest that functional expression of DFF is not sufficient for the induction of DNA fragmentation in Molt-4 cells. PMID- 11872179 TI - Structural change of ribosomes during apoptosis: degradation and externalization of ribosomal proteins in doxorubicin-treated Jurkat cells. AB - Changes in the amount and localization of human ribosomal proteins during apoptosis were determined. When total lysates of Jurkat cells undergoing apoptosis induced by doxorubicin were analyzed by Western blotting, degradation of three ribosomal proteins, S18, L5, and L14, was detected at 48 h after the induction of apoptosis. Decreases in the amounts of these three ribosomal proteins were also observed in ribosome-enriched fractions. These changes were partly abolished by the addition of the pan-caspase inhibitor z-VAD-fmk. Moreover, formation of the 80S ribosome complex appeared to be inhibited at 48 h after apoptosis induction. On the other hand, the rate of protein synthesis, assessed by measuring the incorporation of [35S]Met into bulk proteins, decreased as early as 12 h after the addition of doxorubicin. These results indicate that changes in the amount of ribosomal proteins and the overall structure of ribosomes in apoptosing cells occur after protein synthesis declines. Finally, analyses by flow cytometry, immunofluorescence, and Western blotting showed that six ribosomal proteins, S15, P0, L5, L6, L36a, and L41, were relocalized and expressed at the cell surface during apoptosis. The above results collectively indicate that ribosomes are structurally altered in apoptotic cells following inactivation of protein synthesis. PMID- 11872178 TI - Oxidative damage due to copper ion and hydrogen peroxide induces GlcNAc-specific cleavage of an Asn-linked oligosaccharide. AB - Cleavage of an asparagine-linked sugar chain by hydrogen peroxide (H2O2) and a copper salt was investigated. Incubation of a 2-aminopyridine (PA)-labeled biantennary sugar chain, GlcNAcbeta1-2Manalpha1-6(GlcNAcbeta1-2Manalpha1 3)Manbeta1-4GlcNAcbeta1-4GlcNAc-PA, with H2O2 and Cu2+ led to formation of four major degradation products. Reversed phase high performance liquid chromatographic analysis coupled with glycosidase digestion indicated that the sugar chain is not randomly degraded but specifically degraded at a GlcNAc residue. Treatment with either of H2O2 or copper alone did not cleave nor degrade the sugar chain to any extent. Electron spin resonance (ESR) spectra obtained using a spin trap reagent were consistent with the generation of OH* or an OH* like radical by the H2O2/copper salt mixture. The addition of ascorbic acid enhanced this radical generation as well as the degradation of the sugar chain. It was also found that H2O2/Cu2+ destroys the N-acetyl group of the monosaccharide GlcNAc, as judged by a decrease in the ultraviolet absorption spectrum of this group. On the other hand, replacement of copper by Fe2+ caused no cleavage of the sugar chain, although comparable levels of the same radical species were generated. Furthermore, spectrophotometric analysis showed that a GlcNAc-containing sugar chain coordinates to copper but not to iron, and, thus, the coordination appears to play an essential role in the degradation of the sugar chain. These findings suggest that coordination of copper ions to GlcNAc residues localizes the generation of a radical, which cleaves the glycosidic linkage, possibly involving alteration of the N-acetyl group, thereby allowing the GlcNAc-specific cleavage. PMID- 11872180 TI - Chronic lumbar spine and radicular pain: pathophysiology and treatment. AB - The lumbar spine forms the foundation and infrastructure of an organic skyscraper equipped with the physiologic capacity to act as a crane for lifting and a crankshaft for walking. Subjected to aging like other "human machinery," the lumbar spine adapts to the wear and tear of gravity and biomechanical loading through structural and neurochemical changes. Many of the changes are maladaptive, resulting in pain, physical and functional disability, and altered neurophysiologic circuitry. Some compensatory reactions are constructive, but others cause more interference with the organism's capacity to cope. A conceptional understanding of the multifaceted structural, biomechanical, biochemical, medical, and psychosocial influences that compose this mix elucidates the complexity of applying effective treatments. PMID- 11872181 TI - Is fibromyalgia a neurologic disease? AB - Fibromyalgia (FM) is characterized by abnormal pain sensitivity in response to diverse stimuli as well as persistent widespread pain and other symptoms such as fatigue and sleep disturbance. Progress has been made in identifying factors that contribute to the etiopathogenesis of abnormal pain sensitivity, but there is no single model of pathophysiology or treatment of FM that has gained wide acceptance among health care professionals. We review the literature on the etiopathogenesis of abnormal pain sensitivity in FM and describe an explanatory model that serves as a source of testable hypotheses in our laboratory. This model posits that interactions of exogenous (e.g., environmental stressors) and endogenous (e.g., neuroendocrine dysfunction) abnormalities in genetically predisposed individuals lead to a final common pathway, i.e., alterations in central nervous system function and neuropeptide production that underlie central sensitization and abnormal pain sensitivity. This model also suggests that efforts to develop and evaluate treatments for FM should focus on interventions with direct or indirect effects on central functions that influence pain sensitivity. PMID- 11872183 TI - AIDS and AIDS-treatment neuropathies. AB - AIDS and AIDS-treatment neuropathies are common in individuals infected with HIV. As patients live longer due to improved antiretroviral therapies, the impact of painful neuropathy on patients' lives may increase. Several antiretroviral medications are known to cause toxic neuropathy in patients with AIDS, but this may be outweighed by the beneficial effects of viral suppression. Current theories on the pathogenesis of AIDS neuropathies include mitochondrial toxicity secondary to gamma-DNA polymerase inhibition and subsequent abnormal mitochondrial DNA synthesis. Treatment of AIDS neuropathies is directed toward relief of symptoms; however, new evidence suggests that aggressive antiretroviral therapy may also be effective. PMID- 11872182 TI - Trigeminal neuralgia and other neuropathic pain syndromes of the head and face. AB - Trigeminal neuralgia is the most common craniofacial pain syndrome of neuropathic origin. Although the diagnosis remains based exclusively on history and symptomatology, modern diagnostic techniques, particularly high-resolution magnetic resonance imaging, provides valuable new insight into the pathophysiology of these cases with additional implications for therapeutic strategies. Other neuropathic syndromes affect the trigeminal nerve and warrant different treatments with varied rates of success. Rarely, neuralgias of other cranial nerves mimic trigeminal neuralgia. Finally, it is imperative to distinguish atypical facial pains from these neuropathic syndromes to avoid unsuccessful therapies. PMID- 11872185 TI - Triptans: are they all the same? AB - The triptans have provided a major advance in the treatment of the pain and disability associated with migraine headache. With seven triptans in use or in clinical development, the clinician is faced with the decision of which triptan to prescribe to the patient with migraine. Although the triptans are pharmacologically similar, they each have unique attributes. This article focuses on the pharmacologic differences between triptans with regard to their pharmacokinetics and drug interactions, and provides some helpful tips on how to optimize migraine treatment with the triptans. PMID- 11872184 TI - Triptans: do they differ? AB - Headache care specialists agree that the introduction of sumatriptan constitutes a major advance in headache therapy, but they differ about whether other triptans offer clinically significant advantages over sumatriptan. This article examines this issue by considering the similarities and differences among triptans. PMID- 11872186 TI - Butalbital-containing agents: should they be banned? No. AB - Butalbital compounds are of proven efficacy in the treatment of tension headache. Decades of experience have established their value in the treatment of other mild to-moderate headaches. Untold numbers of people rely on these medications as their drug of choice or use them when vasoconstrictors, opioids, or nonsteroidal anti-inflammatory agents are contraindicated. The medications are cost-effective with only occasional and minor immediate adverse effects. Their overuse may cause the evolution of episodic primary headaches to chronic daily headaches; however, removal of these agents from the market would reduce the chronic daily headache in the general population by a small fraction of 1%. PMID- 11872188 TI - Do over-the-counter medications help the physician manage migraine headache? AB - The use of over-the-counter-medication in migraine treatment is commonplace. They are often used in addition to physician-prescribed medications and may be used without the full knowledge of the physician. Although considerable evidence suggests their efficacy in migraine treatment, they are rarely used to maximal levels of efficacy and safety. A review of the evidence for these treatments is important for cost effective, well-tolerated, and specific treatment of migraine headache. PMID- 11872187 TI - Should butalbital-containing analgesics be banned? Yes. AB - In the United States analgesic-overuse headache is often caused by butalbital containing analgesics. These agents can cause physical and psychological dependency, and dangerous withdrawal syndromes. Butalbital-containing analgesics have already been banned in several European countries. They are proven effective in tension-type headache, but not in migraine; there are many alternative treatments for migraine and tension-type headache. In the 20 years since analgesic overuse headache was widely recognized, butalbital overuse has remained distressingly common. It is time to ban these agents. PMID- 11872190 TI - Leptin and reproduction: a review. AB - OBJECTIVE: To review recent advances in understanding the role of leptin in the physiology and pathophysiology of reproduction, with a focus on relevant clinical situations. DESIGN: A MEDLINE computer search was performed to identify relevant articles. RESULT(S): Leptin, an adipocyte hormone important in regulating energy homeostasis, interacts with the reproductive axis at multiple sites, with stimulatory effects at the hypothalamus and pituitary and inhibitory actions at the gonads. More recently, leptin has been shown to play a role in other target reproductive organs, such as the endometrium, placenta, and mammary gland, with corresponding influences on important physiologic processes such as menstruation, pregnancy, and lactation. As a marker of whether nutritional stores are adequate, leptin may act in concert with gonadotropins and the growth hormone axis to initiate the complex process of puberty. Conditions in which nutritional status is suboptimal, such as eating disorders, exercise-induced amenorrhea, and functional hypothalamic amenorrhea, are associated with low serum leptin levels; and conditions with excess energy stores or metabolic disturbances, such as obesity and polycystic ovarian syndrome, often have elevated serum or follicular fluid leptin levels, raising the possibility that relative leptin deficiency or resistance may be at least partly responsible for the reproductive abnormalities that occur with these conditions. CONCLUSION(S): Leptin may act as the critical link between adipose tissue and the reproductive system, indicating whether adequate energy reserves are present for normal reproductive function. Future interventional studies involving leptin administration are expected to further clarify this role of leptin and may provide new therapeutic options for the reproductive dysfunction associated with states of relative leptin deficiency or resistance. PMID- 11872189 TI - Do over-the-counter medications for migraine hinder the physician? AB - The availability of many over-the-counter drugs that were formerly prescription medications enable patients with migraine to self-medicate easily and delay entry into the appropriate medical management. The potential for adverse effects, drug interactions, and analgesic rebound headaches can often be complications that hinder treatment. Over-the-counter products force the patient to employ a less effective step-care approach as opposed to evidence-based guidelines. PMID- 11872191 TI - Raloxifene revisited. PMID- 11872192 TI - The National Regional Advisory Council practice survey for 2000. AB - A practice survey for the year 2000 was conducted through the National Regional Advisory Council. Each respondent was asked to supply detailed information on diagnostic and therapeutic procedures in the first 20 consecutive patients seen in that year with follow-up for 1 year or until an ongoing pregnancy was reached. The results show interesting patterns of geographic differences in treatment of the infertile couple. In general, American Society for Reproductive Medicine guidelines for practice were followed. PMID- 11872193 TI - A computerized decision support system for ovarian stimulation by gonadotropins. AB - OBJECTIVE: To evaluate the effectiveness of a computerized decision support system for ovarian stimulation with gonadotropins. DESIGN: Retrospective and prospective randomized studies. SETTING: Private and university teaching hospital. PATIENT(S): Women undergoing ovarian stimulation to treat infertility. MAIN OUTCOME MEASURE(S): Pregnancy rate. RESULT(S): In the retrospective study, computer-generated decisions were compared with clinicians' decisions in 118 stimulated cycles in 53 patients. In 90% of cases, the choice of FSH regimens and adjustments to dosages were identical. In the prospective study, the computer generated decisions achieved a pregnancy rate per cycle of 18% (15 of 82 cycles), compared with 16% (13 of 82 cycles) achieved by clinicians. CONCLUSION(S): A computerized decision making system was as effective as skilled clinicians in achieving pregnancy by using ovarian stimulation with FSH. PMID- 11872194 TI - Computer-assisted controlled ovarian hyperstimulation: where are the assisted reproductive technologies? PMID- 11872196 TI - Early follicular serum mullerian-inhibiting substance levels are associated with ovarian response during assisted reproductive technology cycles. AB - OBJECTIVE: To test the hypothesis that the concentration of early follicular phase serum mullerian-inhibiting substance (MIS) is associated with ovarian response in women undergoing ovulation induction in preparation for assisted reproductive technology (ART). DESIGN: Retrospective analysis of frozen day 3 serum samples. SETTING: Academic ART program. PATIENT(S): One sample of frozen day 3 serum from women with < or = 6 retrieved oocytes (n = 28) compared with women with > or = 11 oocytes retrieved (n = 79) in preparation for IVF. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Comparison of day 3 serum MIS levels between two groups of women. Other comparisons included maximum serum E(2) concentrations, number of retrieved oocytes, and percentage of mature oocytes between groups. RESULT(S): Mean serum MIS concentrations were 1.0 +/- 0.4 ng/mL compared with 2.5 +/- 0.3 ng/mL, or more than a 2.5-fold greater serum concentration of MIS in the group with > or = 11 oocytes retrieved compared with in the group with < or = 6 retrieved oocytes. CONCLUSION(S): These data demonstrate an association between early follicular phase serum MIS and the number of retrieved oocytes. Higher day 3 serum MIS concentrations were associated with greater number of retrieved oocytes. PMID- 11872195 TI - Increased prevalence of thrombophilia among women with severe ovarian hyperstimulation syndrome. AB - OBJECTIVE: To determine the prevalence of markers of thrombophilia in women hospitalized for severe OHSS. DESIGN: Prospective study. SETTING: Academic research center. PATIENT(S): Women undergoing induction of ovulation complicated by severe OHSS (n = 20) and women undergoing induction of ovulation without development of severe OHSS (n = 41). INTERVENTION(S): Blood samples to test for markers of thrombophilia were obtained during the luteal phase of the treatment cycle. MAIN OUTCOME MEASURE(S): Blood samples were analyzed for markers of thrombophilia, such as plasma levels of antithrombin, protein S and protein C, antiphospholipid antibodies, the factor V Leiden mutation, and 677T polymorphism in the 5,10 methyltetrahydrofolate reductase (MTHFR 677T) gene. RESULT(S): Seventeen of 20 patients with severe OHSS (85%) and 11 of 41 controls (26.8%) had one or more positive markers of thrombophilia. Of the women with severe OHSS, 6 had a decreased antithrombin level, 8 had decreased levels of protein S, 7 were homozygous for the MTHFR 677T mutation, 1 was heterozygous for the factor V Leiden mutation, and 5 had antiphospholipid antibodies. Eight women with OHSS and no controls had more than one positive marker of thrombophilia. CONCLUSION(S): The prevalence of thrombophilia is increased in women with severe OHSS. These findings suggest that prophylactic screening for this disorder and possible use of heparin prophylaxis for thromboembolic phenomena should be considered in these patients. PMID- 11872197 TI - Comparison of cryopreservation outcome with gonadotropin-releasing hormone agonists or antagonists in the collecting cycle. AB - OBJECTIVE: To compare the pregnancy rates of frozen-thawed 2-pronucleate (2PN) oocytes obtained either in a long protocol or in an antagonist protocol and ovarian stimulation with either human menopausal gonadotropin (hMG) or recombinant follicular stimulating hormone (recFSH). DESIGN: Retrospective data analysis. SETTING: Academic infertility center. PATIENT(S): Three hundred forty two infertile couples who underwent a transfer of cryopreserved 2PN oocytes. INTERVENTION(S): hMG (n = 194) or recFSH (n = 92) in a long protocol or hMG (n = 16) or recFSH (n = 40) stimulation under pituitary suppression with the GnRH antagonist Cetrotide was used. The 2PN oocytes were transferred after endometrial preparation using E(2) valerate and vaginal progesterone (Crinone 8% vaginal gel). MAIN OUTCOME MEASURE(S): Implantation, pregnancy, and abortion rates. RESULT(S): Implantation rates in the freeze-thaw cycles were 5.6% (hMG) and 3.8% (recFSH) with 2PN oocytes from the long protocol and 7% from the antagonist cycles, irrespective of whether hMG or recFSH was used. Pregnancy rates were similar independent of whether they resulted from the long-protocol cycles with hMG (15.4%) and recFSH (13.1%) or from the antagonist protocol cycles with hMG (25.0%) and recFSH (17.5%). CONCLUSION(S): The potential to implant is independent of the gonadotropin-releasing hormone analogue and gonadotropin chosen for the collection cycle when previously cryopreserved 2PN oocytes were replaced after thawing in the cleavage stage. PMID- 11872198 TI - Effects of transdermal hormone replacement therapy on levels of soluble P- and E selectin in postmenopausal healthy women. AB - OBJECTIVE: To study the adhesion molecule pattern in postmenopausal women who were not receiving hormone replacement therapy (HRT), HRT users, and fertile women. DESIGN: Case-control study. SETTING: Second University of Naples, Naples, Italy. PATIENT(S): Fifty healthy naturally postmenopausal women and 20 fertile women. INTERVENTION(S): Twenty-six women received no HRT and 24 received continuous transdermal 17 beta-estradiol, 0.05 mg/d, plus oral acetate nomegestrol, 5 mg/d. MAIN OUTCOME MEASURE(S): Levels of the soluble forms of intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), E-selectin, and P-selectin. RESULT(S): Women who did not received HRT showed a trend toward higher levels of soluble E-selectin and had significantly higher levels of soluble P-selectin than did fertile women. Levels of soluble E selectin and soluble P-selectin were significantly lower in HRT users than in nonusers. Levels of VCAM-1 levels were significantly higher in HRT users than in fertile women, but no significant differences in CAM concentrations were found between the other groups. CONCLUSION(S): Menopause may lead to increased levels of soluble E- and soluble P-selectin, whereas long-term HRT is associated with lower selectin concentrations. This suggests that HRT may have a beneficial effect on endothelial function. PMID- 11872199 TI - Effects of hormone replacement therapy and methylenetetrahydrofolate reductase polymorphism on plasma folate and homocysteine levels in postmenopausal Japanese women. AB - OBJECTIVE: To evaluate the relationships among the methylenetetrahydrofolate reductase (MTHFR) polymorphism, plasma folate, total homocysteine (Hcy) levels, lipids, and the reduction of Hcy levels resulting from hormone replacement therapy (HRT). DESIGN: Clinical study. SETTING: Outpatient department of obstetrics and gynecology in a general hospital. PATIENT(S): Two hundred seventeen postmenopausal Japanese women. INTERVENTION(S): Of the 217 women, 172 patients were under continuous treatment with oral conjugated equine estrogen and medroxyprogesteron acetate. MAIN OUTCOME MEASURE(S): Fasting Hcy, folate, methionine, lipids, and apolipoproteins were measured before and after 3 months of HRT. RESULT(S): The plasma Hcy concentration was significantly higher in the low folate than in the high-folate group only in patients with the homozygous (T/T) mutant. Plasma Hcy concentrations were significantly correlated with age (R = 0.64, P=.02) or years since menopause (R = 0.73, P=.02) only in the low-folate group with T/T. The plasma Hcy concentration decreased significantly in all genotypes after 3 months of HRT, but the levels of serum folate and methionine remained unchanged. CONCLUSION(S): The MTHFR polymorphism was associated with a higher Hcy concentration, and this association was related to the serum folate level. Hormone replacement therapy reduced the plasma Hcy concentration independently of the MTHFR polymorphism. PMID- 11872200 TI - Estrogen raises the sweating threshold in postmenopausal women with hot flashes. AB - OBJECTIVE: To determine if estrogen ameliorates hot flashes by raising the core body temperature sweating threshold, by reducing core body temperature fluctuations, and/or by reducing sympathetic activation (as measured by plasma 3 methoxy-4-hydroxyphenylglycol). DESIGN: Laboratory physiological study. SETTING: University medical center. PATIENT(S): Twenty-four healthy postmenopausal women reporting frequent hot flashes. INTERVENTION(S): Participants were randomly assigned, in double-blind fashion, to receive 1 mg/d 17beta-estradiol orally or placebo for 90 days. MAIN OUTCOME MEASURE(S): Core body temperature, core body temperature fluctuations, mean skin temperature, sternal sweat rate, laboratory hot flash counts (sternal skin conductance), plasma 3-methoxy-4 hydroxyphenylglycol. RESULT(S): The E(2) group had significant increases in plasma E(2) (8 +/- 2 vs. 132 +/- 22 pg/mL) and core body temperature sweating threshold (37.98 +/- 0.09 vs. 38.14 +/- 0.09 degrees C) and decreases in plasma FSH (58.8 +/- 8.9 vs. 40.1 +/- 7.6 mIU/mL) and hot flashes (1.4 +/- 0.5 vs. 0.6 +/- 0.6). These changes did not occur in the placebo group. There were no significant changes in any other measure. CONCLUSION(S): E(2) ameliorates hot flashes by raising the core body temperature sweating threshold, but does not affect core temperature fluctuations or plasma 3-methoxy-4-hydroxyphenylglycol. PMID- 11872201 TI - Effects of folic acid and zinc sulfate on male factor subfertility: a double blind, randomized, placebo-controlled trial. AB - OBJECTIVE: To study the effects of folic acid and zinc sulfate treatment on semen variables in fertile and subfertile men. DESIGN: Double-blind, placebo-controlled interventional study. SETTING: Two outpatient fertility clinics and nine midwifery practices in The Netherlands. PARTICIPANT(S): One hundred eight fertile and 103 subfertile men. INTERVENTION(S): Both groups were randomly assigned to receive one of four treatments for 26 weeks: folic acid and placebo, zinc sulfate and placebo, zinc sulfate and folic acid, and two placebos. Folic acid was given at a daily dose of 5 mg, and zinc sulfate was given at a daily dose of 66 mg. MAIN OUTCOME MEASURE(S): Before and after treatment, standardized semen and blood samples were obtained for determinations of sperm concentration, motility, and morphology according to World Health Organization guidelines; semen morphology according to strict criteria; and blood folate and zinc concentrations. Effects of the four interventions were evaluated separately in subfertile and fertile men. RESULT(S): Subfertile men demonstrated a significant 74% increase in total normal sperm count and a minor increase of 4% abnormal spermatozoa. A similar trend was observed in fertile men. Pre-intervention concentrations of folate and zinc in blood and seminal plasma did not significantly differ between fertile and subfertile men. CONCLUSION(S): Total normal sperm count increases after combined zinc sulfate and folic acid treatment in both subfertile and fertile men. Although the beneficial effect on fertility remains to be established, this finding opens avenues of future fertility research and treatment and may affect public health. PMID- 11872202 TI - Alternatively spliced variants of the follicle-stimulating hormone receptor gene in the testis of infertile men. AB - OBJECTIVE: To investigate whether or not alternatively spliced variants of the FSH receptor gene occur in human testis and whether the presence of the splicing variants is associated with spermatogenic defects and serum FSH concentration in infertile men. DESIGN: A prospective case control study. SETTING: An IVF clinic and infertility laboratory at a university hospital. PATIENT(S): Forty-three infertile patients undergoing testicular biopsy. INTERVENTION(S): Total RNA was extracted from the testicular tissues and used for reverse transcriptase polymerase chain reaction (RT-PCR). MAIN OUTCOME MEASURE(S): Expression pattern was analyzed by nested RT-PCR using primers designed to amplify a fragment of FSH receptor gene. PCR products of splicing variants were cloned and sequenced. RESULT(S): The PCR products showed three kinds of additional bands corresponding to alternatively spliced isoforms of the FSH receptor gene. Exon 9 deleted variant was detected in all patients and inclusion variant of small extra exon was detected in 64% (9/14) of the patients with normal spermatogenesis and 34% (10/29) of the patients with spermatogenic defects. The presence of inclusion variant was not significantly associated with spermatogenic defects but was associated with a low level of serum FSH. On the other hand, exon 6 deleted variant was detected in only one patient having a high level of FSH concentration (30 IU/L) and Sertoli cell only syndrome. CONCLUSION(S): We identified three different types of alternatively spliced variants of the human FSH receptor. However, it is not clear whether or not there is an association between three variants and spermatogenic defects. PMID- 11872203 TI - Cumulative probability of live birth after three in vitro fertilization/intracytoplasmic sperm injection cycles. AB - OBJECTIVE: To assess the probability of live birth after three available in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) cycles. DESIGN: Retrospective, observational study. SETTING: University hospital. PATIENT(S): Nine hundred seventy-four couples who started their first conventional IVF or ICSI cycle between January 1996 and December 1997. A total of 1985 stimulated cycles were initiated. INTERVENTION(S): Analysis of the cumulative live birth rate using the life-table approach with and without taking dropouts into account. MAIN OUTCOME MEASURE(S): Cumulative live birth rate. RESULT(S): The overall cumulative live birth rate after three completed stimulated cycles (including freezing/thawing cycles) was 65.5% with an "optimistic" approach. For the "realistic" and "pessimistic" approaches the corresponding figures were 63.1% and 55.5%, respectively. Unexpectedly, 65% of couples not achieving a live birth interrupted the full treatment program of three cycles. CONCLUSION(S): The cumulative live birth rate gives the couple a more accurate prognosis of achieving a live birth after IVF/ICSI than the statistics usually provided. With the "realistic" estimation, 63% of the couples achieved childbirth after three available conventional IVF or ICSI cycles. Further studies are required to investigate the high drop-out rate. PMID- 11872204 TI - Apoptosis in human cumulus cells in relation to zona pellucida thickness variation, maturation stage, and cleavage of the corresponding oocyte after intracytoplasmic sperm injection. AB - OBJECTIVE: To assess the degree of apoptosis in the cumulus cells and the variation of the zona pellucida and the maturity and fertilization of the corresponding oocyte. DESIGN: Retrospective study. SETTING: Private fertility clinic. PATIENT(S): Fifty couples undergoing ICSI. INTERVENTION(S): ICSI. MAIN OUTCOME MEASURE(S): Correlation between apoptosis in the cumulus cells and the zona pellucida thickness variation, maturation stage, fertilization rate, and embryo score. RESULT(S): This study demonstrated no correlation between apoptosis in cumulus cells and the thickness and variation of the zona pellucida in oocytes and embryos. The incidence of apoptosis was significantly higher in cumulus cells from empty zona pellucidas and germinal vesicle stage and metaphase I oocytes compared with metaphase II oocytes. Non-fertilized metaphase II oocytes showed significantly higher incidence of apoptosis compared with fertilized metaphase II oocytes. There was a correlation between embryo score and the zona pellucida thickness variation. CONCLUSION(S): Apoptosis in cumulus cells had no impact on the zona pellucida thickness and variation in oocytes and embryos. The zona pellucida thickness variation was positively correlated to good embryo score. A higher degree of apoptosis was seen in cumulus cells from immature oocytes compared with mature oocytes. Furthermore, apoptosis in cumulus cells impaired the fertilization rate of metaphase II oocytes after ICSI. PMID- 11872205 TI - Vasectomy reversal performed 15 years or more after vasectomy: correlation of pregnancy outcome with partner age and with pregnancy results of in vitro fertilization with intracytoplasmic sperm injection. AB - OBJECTIVE: To document a contemporary series of vasectomy reversals performed in men 15 years or more after vasectomy and to correlate the results with spousal age and results of ICSI for obstructive azoospermia. SETTING: University referral center for male infertility. DESIGN: Retrospective analysis of a single surgeon's experience compared with reported ICSI results. PARTICIPANT(S): One hundred seventy-three men who had vasectomy reversal 15 years or more after vasectomy. INTERVENTION(S): Reversal of vasectomy by vasovasostomy or epididymovasostomy. MAIN OUTCOME MEASURE(S): Correlation of pregnancy results after vasectomy reversal with spousal age and published ICSI results. RESULT(S): Pregnancy rates for the intervals of 15-19 years, 20-25 years, and >25 years after vasectomy were 49%, 39%, and 25%, respectively. For spousal age <30 years, 30--35 years, 36-40 years, and >40 years, pregnancy rates were 64%, 49%, 32%, and 28%, respectively. The overall pregnancy rate was 43%, which is similar to the pregnancy rate of 40% for ICSI in obstructive azoospermia. Sixty-two percent of the men required a unilateral or bilateral epididymovasostomy. CONCLUSION(S): Spousal age is an important predictive factor after vasectomy reversal among men who have reversal 15 years or more after vasectomy. Pregnancy rates after vasectomy reversal compare favorably with those obtained with ICSI. PMID- 11872206 TI - Metformin therapy throughout pregnancy reduces the development of gestational diabetes in women with polycystic ovary syndrome. AB - OBJECTIVE: To assess whether metformin safely reduced development of gestational diabetes in women with the polycystic ovary syndrome (PCOS). DESIGN: Prospective and retrospective study. SETTING: Outpatient clinical research center. PATIENT(S): The prospective study included 33 nondiabetic women with PCOS who conceived while taking metformin and had live births; of these, 28 were taking metformin through delivery. The retrospective study included 39 nondiabetic women with PCOS who had live birth pregnancies without metformin therapy. INTERVENTION(S): Metformin, 2.55 g/d, throughout pregnancy in women with PCOS. MAIN OUTCOME MEASURE(S): Development of gestational diabetes in women with PCOS. RESULT(S): Before metformin therapy, after covariance adjustment for age, the two cohorts did not differ in height, weight, basal metabolic index, insulin, insulin resistance, or insulin secretion. Both cohorts had high fasting insulin, were insulin resistant, and had high insulin secretion. Among the 33 women who received metformin, gestational diabetes developed in 1 of 33 (3%) pregnancies versus 8 of 12 (67%) of their previous pregnancies without metformin. Among the 39 women who did not take metformin, gestational diabetes developed in 14 of 60 (23%) pregnancies. When all live births were combined, gestational diabetes occurred in 22 of 72 pregnancies (31%) in women who did not take metformin versus 1 of 33 pregnancies (3%) in those who took metformin. With gestational diabetes as the response variable and age at delivery and treatment group (metformin or no metformin) as explanatory variables, the odds ratio for gestational diabetes in women with metformin versus without metformin was 0.093 (95% CI: 0.011 to 0.795). With gestational diabetes in 93 pregnancies as the response variable and age at delivery and treatment group (metformin no metformin) as explanatory variables, the odds ratio of gestational diabetes in pregnancies in women taking metformin versus without metformin was 0.115 (95% CI: 0.014 to 0.938). CONCLUSION(S): In PCOS, use of metformin is associated with a 10-fold reduction in gestational diabetes (31% to 3%). It also reduces insulin resistance and insulin secretion, thus decreasing the secretory demands imposed on pancreatic beta-cells by insulin resistance and pregnancy. PMID- 11872208 TI - Clinical outcome of day 2 versus day 5 transfer in cycles with one or two developed embryos. AB - OBJECTIVE: To determine whether extended culture of embryos to blastocysts has any benefit in cycles with only one or two created embryos. DESIGN: Retrospective analysis of cycles comparing outcomes of day 2 and day 5 transfers. Our day 2 group was from the year 1999 and our day 5 group, from the year 2000. SETTING: Assisted reproductive technology program of a teaching hospital. PATIENT(S): All patients, irrespective of age, who had developed one or two embryos. INTERVENTION(S): Stimulated IVF, intracytoplasmic sperm injection, or testicular sperm extraction and intracytoplasmic sperm injection cycles with 2-day culture in universal IVF medium (n = 133) or 5-day culture in BlastAssist media (MediCult, Jyllinge, Denmark; n = 132). MAIN OUTCOME MEASURE(S): Pregnancy, implantation, and take-home baby rates. RESULT(S): In the groups of 2-day and 5 day culture, embryo transfer was performed in 98% and in 57% of cycles, respectively. However, the total implantation rate per created embryo (18% vs. 18%), the pregnancy rate per cycle (23% vs. 21%), and the take-home baby rate (69.4% vs. 71.4%) did not differ between the day 2 and day 5 groups. CONCLUSION(S): Extended culture of embryos does not improve or decrease their capacity for implantation but only allows for better selection and is therefore not necessary in cycles with fewer than three embryos. PMID- 11872207 TI - Effect of hypertension therapy with the angiotensin-converting enzyme inhibitor lisinopril on hyperandrogenism in women with polycystic ovary syndrome. AB - OBJECTIVE: To investigate the effect of an angiotensin-converting enzyme inhibitor, lisinopril, on serum androgen and sex-hormone-binding globulin (SHBG) levels in hypertensive women with polycystic ovary syndrome (PCOS). DESIGN: Prospective, observational study. SETTING: Infertility Clinic of the Sani Konukoglu Hospital, Gaziantep, Turkey. PATIENT(S): Ten hypertensive women with PCOS. INTERVENTION(S): Lisinopril, 10 mg/day, for 4 weeks. MAIN OUTCOME MEASURE(S)): Serum levels of gonadotropins, DHEAS, total T, free T, 17 alpha hydroxyprogesterone (17-OHP), androstenedione, E2, SHBG, TSH, and PRL were determined. RESULT(S): Hypertension treatment with lisinopril (10 mg/day for 4 weeks) resulted in a statistically significant decrease in serum free T levels. However, there was no difference in the SHBG levels. CONCLUSION(S): Use of lisinopril, an angiotensin-converting enzyme inhibitor, results in decreased free T levels independently of SHBG. It may affect the free T levels by affecting the ovarian renin-angiotensin system. PMID- 11872209 TI - Attitudes on access to services at assisted reproductive technology clinics: comparisons with clinic policy. AB - OBJECTIVE: To determine the opinions of assisted reproductive technology (ART) clinic directors on access-to-services issues and to compare these opinions with policy at ART clinics. DESIGN: Survey sent to ART clinic directors. SETTING: Academic medical center, university-based ethics institute. PATIENT(S): None. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Assisted reproductive technology directors' opinions about access to services are related to specific clinic policies. Access issues examined include patient attributes (marital status, age, sexual preference), patient behaviors (substance abuse, psychiatric history, child abuse), number of cycles, and types of procedures offered (surrogacy, cloning, ova cryopreservation, ART for HIV-positive individuals). RESULT(S): Provider opinion was more restrictive than clinic policy in 19 of 20 categories measured. The largest difference between opinion and policy was seen on the question of setting restrictions as to age of the male partner. About 20% of directors think that surrogacy for convenience should be allowed: about 10% of directors think cloning of human beings should be offered. CONCLUSION(S): The opinions of ART directors on access to services are more conservative than clinic policies. Contributing factors may include respect for patient autonomy; fear of litigation; policy that reflects a continuum of beliefs among providers; and economic pressure to offer a broad range of procedures. PMID- 11872210 TI - Estradiol down-regulates MCP-1 expression in human coronary artery endothelial cells. AB - OBJECTIVE: To determine whether estrogen down-regulates MCP-1 in vascular endothelial cells. DESIGN: A prospective comparative study. SETTING: Academic research environment. PATIENT(S): Human umbilical vein endothelial cells (n = 3) and human coronary artery endothelial cells (n = 3) obtained from females. INTERVENTION(S): Human umbilical vein endothelial cells (HUVEC) and human coronary artery endothelial cells (HCAEC) were grown to preconfluence. Then, they were treated with various concentrations of estradiol (10(-11) M to 10(-7) M) as well as raloxifene (10(-7) M) and tamoxifen (10(-7) M). MCP-1 in culture media was quantified using an enzyme-linked immunosorbent assay (ELISA). Cellular ribonucleic acid (RNA) was extracted and Northern blots were hybridized with an oligonucleotide probe complementary to a specific sequence of MCP-1 mRNA. MAIN OUTCOME MEASURE(S): MCP-1 protein and mRNA. RESULT(S): Estrogen treatment did not change MCP-1 expression in HUVEC. On the other hand, in HCAEC, estradiol induced a 30% decrease in mRNA expression and resulted in dose-dependent inhibition of MCP-1 production as detected by ELISA. Raloxifene and tamoxifen also resulted in inhibition of MCP-1 mRNA and protein expression. CONCLUSION(S): Our findings suggest that one of the mechanisms by which estrogen down-regulates atherosclerosis is by suppressing vascular MCP-1 expression, resulting in decreased macrophage recruitment. PMID- 11872211 TI - Effects of exogenous estrogen on uterine leukocyte recruitment. AB - OBJECTIVE: To determine the role of estrogen in leukocyte recruitment to the human endometrium. DESIGN: Prospective, controlled in vivo study. SETTING: Academic research laboratory. PATIENT(S): Ten patients presenting for donor oocytes. INTERVENTION(S): Endometrial biopsies for the evaluation of leukocyte populations were collected from perimenopausal women in two consecutive regulated cycles who were given two different regimens of estrogen with identical progesterone treatment. MAIN OUTCOME MEASURE(S): Immunohistochemical identification of endometrial leukocyte populations and relative levels of expression of three chemokine genes. RESULT(S): The total uterine leukocyte population increased significantly when the women received oral estrogen, which resulted in higher serum estrogen levels. This rise in leukocytes was due to a significant increase in both the uterine natural killer cells and the macrophage populations. T-cell numbers did not change relative to circulating estrogen levels. The relative abundance of mRNA from three chemokines was also determined. No changes were found in the expression of M-CSF or MCP-1. Interleukin 8 decreased in glands relative to estrogen levels. CONCLUSION(S): These data demonstrate that changes in circulating levels of estrogen can regulate the recruitment of bone marrow-derived cells to the uterine endometrium; however, the mechanism whereby that occurs remains elusive. PMID- 11872212 TI - Leuteinizing hormone responses to leuprolide acetate discriminate between hypogonadotropic hypogonadism and constitutional delay of puberty. AB - OBJECTIVE: To assess if leuprolide acetate stimulation discriminates between hypogonadotropic hypogonadism (HH) and constitutional delay of puberty (CDP) in males. DESIGN: Case-control study. SETTING: Patients attending an academic research environment. PATIENTS: Only male patients were studied: 10 with HH (group 1, age 16.5 +/- 6.0 years), 8 prepubertal with CDP (group 2, age 14.3 +/- 1.2 years), 6 healthy prepubertal (group 3, age 9.5 +/- 3.3 years), and 8 healthy late-pubertal (group 4, age 15.1 +/- 3.1 years). INTERVENTION(S): Blood samples were obtained after an overnight fast. Leuprolide acetate was then administered SC, and blood samples were drawn at 0, 30, 60, 120, 180 minutes, and 6 and 24 hours after stimulation. MAIN OUTCOME MEASURE(S): Clinical follow-up evaluations of data and serum levels of LH, FSH, 17-hydroxyprogesterone, and testosterone. RESULT(S): Basal LH levels were similar in groups 1 through 3 and differed significantly from those in group 4. Peak serum LH levels were significantly higher in CDP compared with HH (8.9 +/- 1.4 vs. 1.4 +/- 0.2 IU/L). Baseline FSH levels were significantly higher only in pubertal boys (versus the HH group); peak levels did not differ among the groups. Basal and peak testosterone levels were significantly higher only in the control pubertal group when compared to the other groups; peak 17-hydroxyprogesterone concentrations were significantly higher in pubertal controls compared with HH and CDP. CONCLUSION(S): Peak LH responses clearly discriminate HH from CDP. Timing for blood sampling should be fixed at 0, 60, 120, 180 minutes after stimulation. PMID- 11872214 TI - Factors associated with the reduction of bone density in patients with gonadal dysgenesis. AB - OBJECTIVE: To correlate bone mineral density (BMD) in women with primary hypoestrogenism caused by 46,XX pure gonadal dysgenesis or Turner's syndrome with age, age at estrogen therapy initiation, length of estrogen use, and body mass index (BMI). DESIGN: Cross-sectional study. SETTING: Academic tertiary-care hospital. PATIENT(S): Thirty-eight women, aged 16 to 35 years (mean, 24.6 years), affected by these genetic disorders. INTERVENTION(S): Measurement of lumbar spine and femoral neck BMD using double x-ray absorptiometry. The results were correlated with the control variables by using Pearson's coefficient of correlation. Variables associated with BMD were evaluated by multiple linear regression analysis. MAIN OUTCOME MEASURE(S): Bone mineral density. RESULT(S): Bone mineral density of the lumbar spine showed that 90% of the women presented osteopenia or osteoporosis. The femoral neck was affected in 55% of these women. The length of estrogen therapy and the BMI showed a positive association with BMD at the lumbar spine and femoral neck, respectively. CONCLUSION(S): Women affected by pure gonadal dysgenesis or Turner's syndrome presented a marked decrease in BMD of the lumbar spine and femoral neck. Medical attention for their diagnosis and early hormone replacement therapy are advised. PMID- 11872213 TI - Potential involvement of hemoglobin and heme in the pathogenesis of peritoneal endometriosis. AB - OBJECTIVE: To test whether hemoglobin may accumulate in the peritoneal cavity in case of endometriosis and to assess whether heme oxygenases (HO), detoxifying heme, are expressed in ectopic endometrium and peritoneal cells. DESIGN: Prospective study involving patients with and without endometriosis. SETTING: Department of gynecology in a university hospital. PATIENT(S): Seventy-six patients undergoing laparoscopy for tubal sterilization or infertility and/or pelvic pain. INTERVENTION(S): Collection of peritoneal fluid (PF), blood samples, and biopsies from endometrium and peritoneum. MAIN OUTCOME MEASURE(S): Measurement of free hemoglobin and its byproduct, total and direct bilirubin, in serum and PF and analysis of HO-1 and HO-2 expression in biopsies by reverse transcription polymerase chain reaction and semiquantitative immunohistochemistry. RESULT(S): Higher levels of hemoglobin were found in the PF of patients with endometriosis. There was no concomitant increase in bilirubin concentrations in the PF, and HO-1 was poorly expressed in peritoneal mesothelium and macrophages. Heme oxygenase-1 and HO-2 were strongly expressed in ectopic endometrium, especially in red lesions. CONCLUSION(S): Our results suggest that heme may be involved in the pathogenesis and/or development of endometriosis and that the HO system, although expressed, might be insufficient to detoxify heme in women with endometriosis. PMID- 11872215 TI - Human oviductin mRNA expression is not maintained in oviduct mucosal cell culture. AB - OBJECTIVE: To determine whether oviduct mucosal cell culture supports the continued production of oviductin, a putative embryotrophic protein. DESIGN: Semiquantitative reverse-transcriptase polymerase chain reaction (RT-PCR) analysis of oviductin messenger RNA (mRNA) expression after oviduct mucosal cell culture. SETTING: University-based obstetrics and gynecology department. PATIENT(S): Ten women undergoing laparoscopy for tubal sterilization or hysterectomy for uterine fibroids. INTERVENTION(S): The mucosal layer was isolated from the oviduct tissue and subjected to routine culture conditions; semiquantitative RT-PCR was performed. MAIN OUTCOME MEASURE(S): The relationship between duration of cell culture and expression of oviductin mRNA. RESULT(S): There was a significant reduction in oviductin mRNA expression after 3 days in culture, with a complete loss after 6 days in 70% of the samples and after 12 days in the remaining 30%. CONLCUSION(S): This is the first study to investigate whether oviductin mRNA continues to be expressed in cultured human oviduct mucosal cells. Our results suggest that oviduct mucosal cells lose their ability to produce oviductin after short-term culture. This method of culture does not appear to be appropriate for a coculture system reliant upon oviductal secretion of oviductin. PMID- 11872216 TI - Common single nucleotide polymorphisms in intron 3 of the calpain-10 gene influence hirsutism. AB - OBJECTIVE: To study three common polymorphisms in intron 3 of the calpain-10 gene (CAPN10) in hyperandrogenic patients. DESIGN: Case-control study. SETTING: Academic hospital. PATIENT(S): Ninety-seven hyperandrogenic patients and 37 healthy controls. INTERVENTION(S): Basal and adrenocorticotropin-stimulated serum samples and genomic DNA samples were obtained during the follicular phase of the menstrual cycle. MAIN OUTCOME MEASURE(S): Genotyping of the UCSNP43, UCSNP44, and UCSNP45 polymorphisms in CAPN10 and serum androgen levels. RESULT(S): Sixteen patients had idiopathic hirsutism, defined as normal serum androgen levels and regular menstrual cycles. Eighty-one hyperandrogenic patients (those presenting with hyperandrogenemic hirsutism or the polycystic ovary syndrome) were analyzed further. UCSNP45 alleles were distributed differently among the study groups. Heterozygosity for the uncommon C allele was increased in patients with idiopathic hirsutism (31.3%) and reduced in hyperandrogenic patients (7.4%) compared with controls (16.2%). The UCSNP44 and UCSNP43 alleles were in linkage disequilibrium, and were distributed equally among patients with idiopathic hirsutism, hyperandrogenism, and controls. However, the uncommon A allele at UCSNP43 was associated with higher hirsutism score (mean [+/- SD], 9.9 +/- 6.8, 12.7 +/- 7.7, and 14.6 +/- 8.2 in GG, GA, and AA participants, respectively). No other differences were observed in clinical and biochemical characteristics, including insulin sensitivity, by CAPN10 variant. CONCLUSION(S): The C allele at the UCSNP45 locus in CAPN10 is associated with idiopathic hirsutism, and UCSNP43 influences the hirsutism score. PMID- 11872217 TI - Effect of hydrosalpinx fluid on secretion of trophoblastic matrix metalloproteinases. AB - OBJECTIVE: To determine if hydrosalpinx fluid affects trophoblastic metalloproteinases (MMPs) secretion. DESIGN: Measurement of the effect of hydrosalpinx and peritoneal fluids (as controls) added to the medium on the MMPs secreted by cytotrophoblastic cells. SETTING: Academic research center. PATIENT(S): Five samples of hydrosalpinx fluid were obtained at the time of ovocyte retrieval. Three samples of peritoneal fluids were collected at laparoscopic sterilization. MAIN OUTCOME MEASURE(S): The concentration and activity of MMP-2 and MMP-9, the concentration of the tissue inhibitor of metalloproteinases (TIMP-1), and the total gelatinolytic activity of the cytotrophoblastic cells were measured in the culture medium. RESULT(S): Hydrosalpinx significantly stimulated MMP-2, MMP-9, and TIMP-1. The net result was a significant stimulation of the total gelatinolytic activity. Peritoneal fluids increased MMP-2, MMP-9, and TIMP-1 concentrations, but the total gelatinolytic activity was not modified. CONCLUSION(S): In contrast to peritoneal fluids, hydrosalpinx stimulates the total gelatinolytic activity of cytotrophoblastic cells. This might indicate that the effect of hydrosalpinx on implantation rates may not be due to an inhibition of the capacity of an embryo to invade the endometrium. However, the stimulatory effect of hydrosalpinx on the net gelatinolytic activity could partly explain the increased incidence of ectopic pregnancies that have been described in the presence of hydrosalpinx. PMID- 11872218 TI - Morphological alterations and DNA fragmentation in oocytes from primordial and primary follicles after freezing-thawing of ovarian cortex in sheep. AB - OBJECTIVE: To evaluate DNA fragmentation in the oocyte of primordial and primary follicles and morphology of these follicles after freezing and thawing of ovarian cortex in sheep using two freezing protocols. DESIGN: Fragmentation of DNA was evaluated by the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end-labeling (TUNEL) technique. SETTING: Fertility clinic in a large university hospital. ANIMALS: Five- to 6-month-old lambs. INTERVENTION(S): Two-millimeter thick slices of hemi-ovary cortex were prepared. MAIN OUTCOME MEASURE(S): Histological structure and DNA fragmentation. RESULT(S): In the frozen fragments, the percentage of morphologically normal follicles was significantly lower for both protocols compared with the case of the control group of fresh fragments. There was no significant difference between the two types of freezing protocols (60.4% +/- 13.2% vs. 68.4% +/- 13.7%). However, the distribution of abnormalities (nucleus, cytoplasm, and nucleus and cytoplasm) was dissimilar. The results of the TUNEL technique for the three groups showed no significant difference, but the percentage of the TUNEL-positive follicles was slightly lower for the frozen fragments for both protocols with respect to the control group. CONCLUSION(S): The freezing and thawing process of the ovarian cortex does not induce fragmentation of the DNA on the oocyte of primary and primordial follicles. PMID- 11872219 TI - Estrogen replacement therapy induces telomerase RNA expression in the macaque endometrium. AB - OBJECTIVE: To evaluate the effects of hormonal therapies on the expression of telomerase RNA (TRNA) in the endometrium of ovariectomized female cynomolgus macaques (Macaca fascicularis). DESIGN: Randomized long-term experimental trial. SETTING: Animal study at an academic research institution. PATIENT(S): Surgically postmenopausal cynomolgus macaques. INTERVENTION(S): Treatments were given in the diet for three years and included conjugated equine estrogens (CEE), CEE + medroxyprogesterone acetate (MPA), and tamoxifen, at clinically relevant doses. MAIN OUTCOME MEASURE(S): Expression of TRNA within the basal glands, basal stroma, superficial glands, and superficial stroma of the endometrium by radiolabeled in situ hybridization. RESULT(S): Conjugated equine estrogens increased glandular TRNA expression, and the addition of MPA decreased this effect. Tamoxifen induced glandular TRNA expression to a lesser degree. Both CEE + MPA and tamoxifen increased stromal TRNA expression. The expression of TRNA in the endometrial glands was always greater than TRNA expression in the stroma. Treatment groups with greater proliferation and progesterone receptor expression also had elevated TRNA; within-group correlations were not significant. No statistically significant difference occurred between the basal and superficial endometrial layers. CONCLUSION(S): These results show for the first time a cell specific hormonal regulation of TRNA in the primate endometrium, with up regulation of TRNA by treatments associated with increased risk of endometrial cancer in women. PMID- 11872220 TI - Laparoscopic excision of ovarian cysts: is the stripping technique a tissue sparing procedure? AB - OBJECTIVE: To determine whether the stripping technique by laparoscopy is a tissue-sparing procedure. DESIGN: Prospective study. SETTING: University hospital. PATIENT(S): Forty-two women, 21 to 35 years of age, who had a unilateral ovarian cyst (26 endometriomas, 7 serous, 6 dermoid, and 3 mucinous cysts). INTERVENTION(S): Laparoscopic excision of ovarian cysts by using the stripping technique. MAIN OUTCOME MEASURE(S): Histologic analysis of the excised specimens was done to evaluate the presence and nature of ovarian tissue adjacent to the cyst wall. RESULT(S): Recognizable ovarian tissue adjacent to the cyst wall was present in 15 of 42 excised specimens (36%). A significant difference was present for endometriomas versus non-endometriosis cysts (ovarian tissue was present in 14 of 26 specimens [54%] vs. 1 of 16 specimens [6%]; P<.005). No specimen showed the normal follicular pattern observed in healthy ovaries. CONCLUSION(S): The stripping technique appears to be a tissue-sparing procedure. In 36% of the cysts, ovarian tissue is excised together with the cyst wall, but this tissue does not show the morphologic characteristics observed in normal ovarian tissue. PMID- 11872221 TI - Healthy twin delivery after day 7 blastocyst transfer coupled with assisted hatching. AB - OBJECTIVE: To report a normal twin delivery after transfer of two fresh day 7 blastocysts. DESIGN: Case report. SETTING: Private infertility clinic. PATIENT(S): A 35-year-old woman with a 6-year history of primary infertility with significant pelvic adhesions. INTERVENTION(S): Review of individual IVF-ET therapy cycle. MAIN OUTCOME MEASURE(S): Full-term delivery after day 7 blastocyst transfer. RESULT(S): During the patient's first IVF-ET cycle, the decision was made to undertake blastocyst transfer after extended culture. No blastocysts had formed until late on day 6, by which time the patient had been hospitalized with a renal stone. Subsequently, on day 7, the patient was asymptomatic and presented for embryo transfer, and after assisted hatching, two expanded blastocysts were transferred to her uterus under ultrasound guidance. After confirmation of implantation of a viable twin, pregnancy was uneventful with no obstetrical complications, and a dizygotic twin was delivered vaginally at 38 weeks of gestation. CONCLUSION(S): Few reports have been made regarding viability of more slowly developing blastocysts; however, this case indicates that blastocysts that did not fully expand until day 7 of extended in vitro culture are still able to implant after superovulation and IVF-ET therapy. Assisted hatching of these embryos may have been beneficial in achieving this successful outcome by hastening the blastocyst hatching, allowing more rapid contact with the endometrium. PMID- 11872222 TI - Uterine rupture after hysteroscopic resection of uterine septum. AB - OBJECTIVE: To describe a patient who underwent hysteroscopic resection of a uterine septum for recurrent miscarriage. The subsequent labor caused uterine rupture. DESIGN: Case report. SETTING: Academic medical center. PATIENT(S): A 37 year-old nullipara with three previous miscarriages. INTERVENTION(S): Resection of the septum by cutting diathermy using the operating hysteroscope. MAIN OUTCOME MEASURE(S): Pregnancy and delivery. RESULT(S): The patient had an uneventful pregnancy and spontaneous labor at 41 weeks. Cesarean section was performed because of suspected fetal distress. During cesarean section, the uterus was ruptured transversely along the fundus at the line of the attachment of the septum. CONCLUSION(S): When fetal distress occurs after previous uterine surgery, uterine rupture must be considered as a possible cause and appropriate treatment is necessary. PMID- 11872223 TI - Successful pregnancy with intrauterine insemination using vasal sperm retrieved by electric stimulation. AB - OBJECTIVE: To determine the feasibility of electric stimulation applied to the epididymis or vas deferens to retrieve vasal sperm. DESIGN: Two case reports. SETTING: Assisted reproduction practice in a hospital. PATIENT(S): Two patients with retrograde ejaculation and severe asthenozoospermia. INTERVENTION(S): Pulse electric simulation was applied to the epididymis or vas deferens to retrieve sperm via a tube cannulated into the vas deferens. After diluting with culture medium, retrieved sperm were used for IUI. MAIN OUTCOME MEASURE(S): The volume of retrieved vasal fluid and motility parameters of the sperm, the achievement of pregnancy, and outcome of pregnancy. RESULT(S): In patient 1, the volumes of vasal fluids and numbers of sperm were 10-40 microL and 31.4-75.9 x 10(6), respectively, during two cycles. Sperm motility was 88.4%--93.2%. His wife became pregnant and was delivered of a healthy infant by IUI during the second cycle. In patient 2, 0.6 x 10(6) sperm with a motility of 70% were retrieved and used for IUI. However, his wife did not become pregnant. CONCLUSION(S): Electric vasal sperm retrieval is a feasible method for collecting sperm from the vas deferens in selected patients with male factor infertility. PMID- 11872224 TI - Conception and spontaneous delivery after total hypophysectomy. AB - OBJECTIVE: To report the first case of pregnancy and spontaneous delivery after total hypophysectomy. DESIGN: A case report. SETTING: University hospital. PATIENT(S): A 34-year-old woman with panhypopituitarism after hypophysectomy in childhood for craniopharyngioma. INTERVENTION(S): Successful fertility treatment followed by sufficient hormone substitution therapy during pregnancy. MAIN OUTCOME MEASURE(S): Clinical variables. RESULT(S): The patient had an uncomplicated spontaneous delivery without any substitution of oxytocin before, during, or after delivery. CONCLUSION(S): Normal pregnancy and delivery are possible in women with lack of pituitary function. Maternal pituitary oxytocin release seems to play a limited role in the onset and progression of labor. PMID- 11872225 TI - Presence of DAZL transcript and protein in mature human spermatozoa. AB - OBJECTIVE: To identify the DAZL transcript and protein location in human spermatozoa. DESIGN: In vitro experiment. SETTING: University-based reproductive genetics laboratory. PATIENT(S): A fertile volunteer. INTERVENTION(S): Reverse transcription-polymerase chain reaction (RT-PCR), Western blot, and immunostaining for DAZL. MAIN OUTCOME MEASURE(S): Expression of DAZL in human spermatozoa. RESULT(S): The DAZL-specific primers yield a 128 bp product in ejaculate. A protein of approximately 33.5 kDa was detected by Western blot analysis. Immunofluorescence staining showed strong homogeneous staining in the midpiece of spermatozoa and weak staining in the principal piece. A speckled-type distribution was found in the head region. CONCLUSION(S): The DAZL transcript and protein are present in human spermatozoa. The roles of DAZL protein in sperm motility and in the sperm-oocyte interaction await further investigation. PMID- 11872226 TI - Ovarian response and in vitro fertilization outcome in patients with reduced ovarian reserve who were stimulated with recombinant follicle-stimulating hormone or human menopausal gonadotropin. PMID- 11872227 TI - IgA in the lumen of the human oviduct is not related to the menstrual cycle but increases during local inflammation. PMID- 11872228 TI - Serum and follicular fluid hormonal levels during ovulation induction. PMID- 11872229 TI - Can luteal phase estradiol levels predict the pregnancy outcome in in vitro fertilization cycles of good responders whose excess embryos yield blastocysts? PMID- 11872230 TI - A critical decision threshold--the temperature. PMID- 11872233 TI - Alloresponses of cord blood cells in primary mixed lymphocyte cultures. AB - The aim of this study was to compare the alloreactive responses against HLA antigens of cord blood cells with those of adult peripheral blood cells. In primary mixed lymphocyte cultures and bulk cell-mediated lympholysis experiments cord blood cells demonstrated significantly decreased proliferation and cytotoxicity. Experiments analyzing the specificity of anti-HLA cytotoxic T lymphocytes (CTL) revealed that cord blood (CB) CTL reacted only partially with third-party cells expressing the stimulating HLA antigens. Lower frequencies of IL-2 producing helper, cytotoxic T-cell precursors and IL-4 producing CB cells were found, whereas the frequencies of IFN-gamma producing cells, as determined by ELISpot experiments, were equivalent to the frequencies of adult IFN-gamma producing cells. Our results imply that, although CB cells have significantly decreased proliferative and cytotoxic alloresponses in bulk mixed lymphocyte cultures, their IFN-gamma production is comparable with that of adult mononuclear cells. Preserved production of IFN-gamma may be a risk factor for the development of graft-versus-host disease and should be taken into consideration when evaluating the possibility for stem cell transplantation with HLA-mismatched CB. PMID- 11872234 TI - Human umbilical cord blood NK T cells kill tumors by multiple cytotoxic mechanisms. AB - Natural killer (NK) T cells are restricted by CD1d and play an important role in the rejection of malignant tumors, but how kill these tumors is unclear. To investigate this, we cultured Valpha24+CD4+ NK T cells in human umbilical cord blood, which was enriched by immunomagnetic beads. In short-term (4 h) cytotoxicity assays, the NK T cells could kill only those targets expressing CD1d. In longer cytotoxicity assays (20 h), however, the NK T cells were able to kill all the tumors, regardless of CD1d expression. When each of the perforin, Fas-FasL, and TNF-alpha cytotoxic mechanisms were blocked, it was apparent that perforin killing dominated in both the short- and long-term assays. In the short term assay, perforin killing required that the targets expressed CD1d, but killing was more efficient if Fas was present because then the Fas-FasL mechanism was also used. Thus, cells that lacked Fas and CD1d and were not killed in the 4 h assay, were instead lysed in 20-h assay through a combination of perforin and TNF-alpha killing. NK T cells can kill tumor targets by perforin, Fas-FasL, and TNF-alpha mechanisms. TNF-alpha killing requires longer contact between effectors and targets, suggesting that TNF-alpha acts by enhancing perforin killing. PMID- 11872235 TI - Functional versus structural matching: can the CTLp test be replaced by HLA allele typing? AB - Human leukocyte antigen (HLA) incompatibilities are the most important immunological barriers to bone marrow transplant success when using unrelated donors. Until recently, standards for donor selection included serological methods for HLA class I antigens and DNA-based typing for HLA class II alleles. In our center cytotoxic T-lymphocyte precursor (CTLp) assays have been an integrated part of the search selection procedure as well. More recently, DNA based typing for HLA class I became available. This allowed us to determine the correlation of CTLp frequencies directed against incompatibilities at the HLA-A, B, and -C locus in 211 donor-recipient pairs. HLA class I incompatibilities are significantly (p < 0.001) associated with high CTLp frequencies. Exceptions did occur, high CTLp frequencies are seen in 14% of the HLA-A, -B, and -C allele matched pairs, whereas in 7% of the pairs mismatched for HLA-A or -B a low CTLp frequency occurred. The successful outcome of transplants performed in the latter cases suggest that the CTLp test can be used as a tool to detect permissible mismatches when no fully matched donor is available. The influence of HLA-C mismatches on the CTLp outcome was less clear. Although in the majority of mismatched pairs (64%) the CTLp frequency was high, in 36% of the pairs the CTLp frequency was low. Analysis of HLA amino acid sequences was performed on the HLA C allele mismatched group. An amino acid difference was always found at five polymorphic positions 97, 99, 113, 114, and 116 situated at the peptide binding groove in the high CTLp frequency group, whereas in the low CTLp frequency group this was observed in only 9 of 17 combinations (p < 0.001). However, this is mainly due to Cw*0303-0304 mismatches. In conclusion, although there is a highly significant correlation between the outcome of the CTLp frequency test and HLA allele class I typing, exceptions occur. It is unclear whether they are all clinically relevant but they certainly provide additional insight in allograft recognition. PMID- 11872236 TI - Mutational analysis of critical residues determining antigen presentation and activation of HLA-DQ0602 restricted T-cell clones. AB - Three different HLA-DQ0602 restricted T-lymphocyte clones (clones 5, 44, and 48) specific for two different Herpes simplex virus type 2 (HSV-2) VP16 peptides were used in a series of proliferation assays with BLS-1 cell lines expressing mutated HLA-DQ0604 molecules as APC. Up to four residues in the peptide-binding region of DQ0604 were replaced by the respective DQ0602 residue. For all three clones, residue beta70 played a crucial role in TCR recognition; beta30 and beta57 were important, although beta86 was less significant. Clone 5 and 48, specific to the HSV-2 VP16 369--379 peptide, responded to the same mutated DQ0604 molecules. Both clones could be stimulated only when the antigen presenting DQ molecule contained the DQ0602-like Gly at position beta70. Stimulation of clone 44, which recognized a different HSV-2 VP16 epitope (VP16 40-50), was less restricted. Molecular homology modeling showed that the beta70Arg of DQ0604 partially covered the peptide around P5/P6. Interactions of beta70 with residues from the antigen peptide and polymorphic residues at positions beta30 and beta57 can modulate this effect. Supported by molecular modeling data, we conclude that the distinct molecular topography of DQ0602 is not contributed by a single residue, but rather the interactions of various polymorphic DQ residues with particular antigenic peptides. PMID- 11872237 TI - Infectious complications in sickle cell disease are influenced by HLA class II alleles. AB - Despite systematic antibiotic therapy, severe infections (septicemia, meningitis, or osteomyelitis) are a major cause of mortality and morbidity in children with sickle cell disease (SCD). In this study, we explored the possibility that polymorphism at the human leukocyte antigen (HLA) locus might constitute an immunogenetic modifying factor to the intrinsic susceptibility to infection in patients with SCD. A cohort of 80 SCD patients living in Paris, 43 with at least one major infectious complication and 37 without infections, were typed for HLA class II loci by polymerase chain reaction-sequence-specific primers (PCR-SSP). We found that significantly more patients without infections carry the HLA class II DRB1*15 specificity than did patients with infections (21.6% in the first group, versus 4.7% in the second group; chi(2) = 10.47, p(c) = 0.01), supporting a protective effect of this allele. Conversely, significantly more patients were found to carry the DQB1*03 specificity within the group of severe infections, supporting a negative effect (34.9% versus 12.2%, chi(2) = 9.41, p(c) = 0.01). These findings suggest a direct involvement of HLA polymorphism in the development of major infections in SCD. Together with previous data on polymorphism of the Fc receptor and of the mannose-binding lectin, they provide evidence for a polygenic immunomodulation of the constitutively increased infectious risk in SCD. PMID- 11872238 TI - Loss or downregulation of HLA class I expression at the allelic level in acute leukemia is infrequent but functionally relevant, and can be restored by interferon. AB - Human leukocyte antigen (HLA) class I expression at the allelic level was analyzed in 397 acute myeloid leukemia (AML) and 186 acute lymphoid leukemia (ALL) using a complement-dependent cytotoxicity assay. Impaired recognition possibly due to HLA downregulation was observed in 2% of the patients with AML and ALL in complete remission, and in 8%-15% in the groups with blasts. In 15 instances of diminished cytotoxicity, leukemic cells and control PHA blasts from the same patients were further analyzed using flow cytometry. In 4/6 ALL and 4/9 AML patients HLA downregulation or complete loss (2 patients) of cell surface expression could be confirmed. No genomic abnormalities were observed. In addition, 12 AML and 13 ALL patients were tested during relapse using flow cytometry. In 1/12 AML patients and 1/13 ALL patients allelic downregulation of cell surface expression was found. In two patients tested, downregulation or loss of cell surface expression of HLA class I antigens corresponded with impaired T cell mediated lysis by HLA restricted cytotoxic T lymphocyte.Treatment of the cells with alpha- or gamma-interferon could restore HLA class I expression and T cell recognition. In conclusion, downregulation of cell surface expression of HLA class I expression at the allelic level in AML and ALL is infrequent but functionally relevant. HLA downregulation was reversible and T-cell recognition could be restored by alpha- or gamma-interferon. PMID- 11872239 TI - B-cell activation and allosensitization after left ventricular assist device implantation is due to T-cell activation and CD40 ligand expression. AB - Left ventricular assist device (LVAD) implantation is frequently complicated by B cell activation and allosensitization, posing a significant risk to successful transplant outcome. This study investigated whether B-cell hyperreactivity and alloantibody production in LVAD recipients involves T-cell dependent pathways. T cell calcium flux and nuclear translocation of NFATc were used to determine states of T-cell activation. Flow cytometry was used to assess human T- and B cell activation after culture with LVAD-derived biomaterial particles. Sera from LVAD recipients and controls were tested for the presence of anti-HLA antibodies, and for soluble CD40 ligand. LVAD-derived biomaterial induced rapid and sustained calcium flux into normal T cells, resulting in calcineurin-dependent nuclear translocation of NFATc. This resulted in increased T-cell expression of CD40 ligand and subsequent B-cell activation, which was reduced by inhibitors of T cell activation (CsA or anti-CD25 mAb) or by anti-CD40 ligand mAb. LVAD recipients demonstrated higher frequencies of anti-HLA antibodies and serum levels of soluble CD40 ligand compared with heart failure controls. The results indicate that exposure of human mononuclear cells to LVAD-derived biomaterial leads to T-cell dependent B-cell activation via CD40--CD40 ligand interaction, and suggest that treatment with calcineurin inhibitors or monoclonal antibodies against either CD25 or CD40 ligand could be effective at preventing B-cell hyperreactivity and allosensitization after LVAD implantation. PMID- 11872240 TI - DRB1*03 diversity and DRB3 associations in five major population groups in the United States. AB - One hundred sixty-one DRB1*03 positive individuals from each of five U.S. population groups (Caucasoids, African Americans, Asians/Pacific Islanders, Hispanics, and Native Americans) were randomly selected from a database of 82,979 individuals. DRB1*03 alleles were identified by polymerase chain reaction sequence-specific oligonucleotide probe typing. A total of six DRB1*03 alleles out of 21 known alleles were detected. DRB1*03011 was the predominant DRB1*03 allele in all populations. Caucasoids were found to be the least diversified; only DRB1*03011 was observed. African Americans carried DRB1*03021 at a high frequency. This allele was observed in three other populations. DRB1*0304 was found in Asians/Pacific Islanders and DRB1*0305, DRB1*0307 and a new allele, DRB1*0316, was found in Hispanics. A subset of individuals was also typed for DRB3 alleles. DRB3*0101, DRB3*0202, and DRB3*0301 were detected and seven DRB1 DRB3 haplotypes were defined. Testing of other individuals not included in the DRB1*03 frequency study identified a variation of a common extended haplotype, A1, B8, DR3, which carries DRB1*0304 and two previously unreported DRB1*03 alleles, DRB1*0311 and *0320, are also described. PMID- 11872242 TI - Inhibition of mitochondrial Na(+)/Ca(2+) exchange by 7-chloro-5-(2-chlorophenyl) 1,5-dihydro-4,1-benzothiazepin-2(3H)-one attenuates free fatty acid efflux in rat cerebral cortex during ischemia-reperfusion injury. AB - We evaluated the effects of 7-chloro-5-(2-chlorophenyl)-1,5-dihydro-4,1 benzothiazepin-2(3H)-one (CGP-37157) (50 muM), a specific inhibitor of mitochondrial Na(+)/Ca(2+) exchange, applied topically onto rat cerebral cortex during ischemia-reperfusion injury. Free fatty acid (FFA) levels in cortical superfusates, withdrawn at 10 min intervals from bilateral cortical windows, were analyzed by high performance liquid chromatography. During a 20 min period of ischemia in control animals, there were significant increases in all FFAs. Following reperfusion, FFA levels remained significantly elevated. Application of CGP-37157 significantly inhibited effluxes of all FFAs during both ischemia and reperfusion. These data indicate that inhibition of mitochondrial Na(+)/Ca(2+) exchange likely prevented the activation of phospholipases that usually occurs following an ischemic insult as evidenced by its attenuation of FFA efflux. PMID- 11872243 TI - Effects of hyperoxia on human sensorimotor cortex activity produced by electrical stimulation of the median nerve: a functional magnetic resonance imaging study. AB - This study investigated the effect of hyperoxia on sensorimotorcortical activity resulting from electrical stimulation of the median nerve, using functional magnetic resonance imaging (fMRI). Nine volunteers underwent stimulation at 5 and 100 Hz while breathing 21% FIO(2) (fraction of inspired oxygen) or 100% FIO(2). fMRI data were correlated with a stimulus predictor curve, transformed into Talairach space and averaged by group. Normoxic (21% FIO(2)) and hyperoxic (100% FIO(2)) sensorimotor activation volumes were compared using Student's t-test. There were no significant differences between the primary somatosensory/primary motor/Brodmann area 40 (SI/MI/Ba40) and secondary somatosensory cortex (SII) activation volumes for normoxia and hyperoxia. (P>0.05). There was no difference between SI/MI/Ba40 and SII activations at 5 and 100 Hz. In contrast to results previously reported for primary visual cortex (V1), hyperoxia did not enhance sensorimotor cortical activation in area SI/MI/Ba40 or SII. These results indicate that there is regional heterogeneity of the fMRI response to hyperoxia in the cerebral cortex. PMID- 11872244 TI - NMDAR-2C and 2D subunits gene expression is induced in brain by neonatal exposure of monosodium L-glutamate to adult rats. AB - Monosodium glutamate (MSG) was administered subcutaneously to male neonate rats, and the effects on N-methyl-D-asparatate (NMDA) subunit receptor types NR2C and NR2D from different brain regions were studied. A semi-quantitative reverse transcription-polymerase chain reaction was used to measure NR2C and NR2D expression levels in the cerebral cortex, hippocampus and striatum. MSG treatment (4 mg/g body weight, on postnatal days 1, 3, 5, and 7) produced an important increase of NR2C and NR2D subunit gene expression levels in the hippocampus and striatum of adults rats. No change was observed in the cerebral cortex. We propose that an early excessive activation of glutamate receptors could modify NMDA subunit expression and its structural composition on postnatal development. This, as part of a compensatory response by an altered neuronal circuitry, mainly in the hippocampus and striatum, suggests that the NMDA receptor could be a determinant factor to modulate the dendritic arrangement and the synaptogenesis. PMID- 11872245 TI - The insecticide imidacloprid is a partial agonist of the nicotinic receptor of honeybee Kenyon cells. AB - The main targets of the insecticide imidacloprid are neuronal nicotinic acetylcholine receptors (nAChRs) within the insect brain. We tested the effects of imidacloprid on ligand-gated ion channels of cultured Kenyon cells of the honeybee, Apis mellifera. Kenyon cells build up the mushroom body neuropils, which are involved in higher order neuronal processes such as olfactory learning. We measured whole-cell currents through nicotinic and gamma-aminobutyric acid (GABA) receptors using patch-clamp techniques. Pressure applications of imidacloprid elicited inward currents, which were irreversibly blocked by alpha bungarotoxin. Imidacloprid was a partial nicotinic agonist, since it elicited only 36% of ACh-induced currents and competitively blocked 64% of the peak ACh induced currents. GABA-induced currents were partially blocked when imidacloprid was coapplied and this block was independent upon activation of nAChRs. Our results identify the honeybee nAChR as a target of imidacloprid and an imidacloprid-induced inhibition of the insect GABA receptor. PMID- 11872247 TI - Overexpression of semicarbazide sensitive amine oxidase in the cerebral blood vessels in patients with Alzheimer's disease and cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy. AB - Semicarbazide sensitive amine oxidase (SSAO) metabolizes oxidative deamination of primary aromatic and aliphatic amines, and, in the brain, it is selectively expressed in blood vessels. SSAO expression is examined, by immunohistochemistry with a purified polyclonal antibody to SSAO from bovine lung, in the brains of subjects with Alzheimer disease (AD; n=10), cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL; n=2), and age-matched controls (n=8). SSAO immunoreactivity is restricted to meningeal and parenchymal blood vessels in control and diseased brains. Yet, a marked and selective increase in SSAO immunoreactivity occurs in association with betaA4 vascular amyloid deposits in patients with AD, and in the vicinity of the typical granular deposits in the blood vessels of gray and white matter in patients with CADASIL. Oxidative deamination of primary aromatic and aliphatic amines by SSAO produces ammonia, hydrogen peroxide and the corresponding aldehyde. Moreover, increased SSAO immunoreactivity is associated with increased Cu/Zn superoxide dismutase 1 expression restricted to abnormal blood vessels in diseased brains. Therefore, it is suggested that increased SSAO expression is a source of oxidative stress in the blood vessel wall in AD and CADASIL. PMID- 11872246 TI - Anandamide inhibits excitatory transmission to rat substantia gelatinosa neurones in a manner different from that of capsaicin. AB - Actions of anandamide (10 microM) were examined on monosynaptic glutamatergic transmission from the periphery to substantia gelatinosa (SG) neurones in adult rat spinal cord slices using the whole-cell patch-clamp technique. In 64% of neurones examined, Adelta-fibre-evoked excitatory postsynaptic currents (EPSCs) were depressed (in amplitude; by 32 +/- 4%, n=21) by anandamide. On the contrary, an inhibitory action on C-fibre-evoked EPSCs was observed in only 31% of neurones tested; this magnitude (17 +/- 3%, n=4) was less than that of Adelta-fibre EPSCs (P<0.05). A cannabinoid-receptor agonist, WIN 55,212-2 (5 microM), exhibited similar actions on the EPSCs. In a neurone with minimal effects of anandamide on C-fibre EPSCs, capsaicin (1 microM) largely depressed the EPSCs (n=3); Adelta fibre EPSCs were little affected by capsaicin. It is concluded that unlike capsaicin, anandamide inhibits more effectively Adelta-fibre than C-fibre mediated excitatory transmission in the SG, possibly through the activation of the cannabinoid receptor. PMID- 11872248 TI - Repeated experience of social defeats increases serotonin transporter and monoamine oxidase A mRNA levels in raphe nuclei of male mice. AB - Serotonin transporter (SERT) and monoamine oxidase A (MAOA) mRNA levels in the raphe nuclei area of the midbrain were measured by the multiplex reverse transcription-polymerase chain reaction method in male mice with repeated experience of social victories (winners) and defeats (losers) in ten daily agonistic confrontations. Experiments revealed enhanced SERT and MAOA mRNA levels in the losers compared with the winners and controls. It has been supposed that SERT and MAOA genes are involved in enhancement of serotonin inactivation in response to the increase of serotonergic activity shown earlier in the losers. A positive correlation between MAOA and SERT mRNA levels in the raphe nuclei area of the midbrain was shown. PMID- 11872249 TI - Glyceraldehyde-3-phospate dehydrogenase is translocated into nuclei through Golgi apparatus during apoptosis induced by 6-hydroxydopamine in human dopaminergic SH SY5Y cells. AB - 6-Hydroxydopamine is a neurotoxin specific to dopamine neurons, and this neurotoxin at 20 muM was confirmed to induce mainly apoptosis in human dopaminergic neuroblastoma SH-SY5Y cells. During the apoptotic process, translocation of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) into nuclei was observed, which is now proposed as an apoptogenic signal in various types of apoptosis. However, it remains to be clarified, how GAPDH is translocated into nuclei. In this paper, GAPDH translocation was followed by fluoro- and electron microscopic observation using antibody against GAPDH. Before the nuclear translocation, a condensed mass of GAPDH protein was detected in the Golgi apparatus, which was identified by 6-([N-(7-nitrobenz-2-oxa-1,3-diazol-4 yl)amino]hexanoyl)-sphingosine, a selective marker of Golgi apparatus. These results suggest that the Golgi apparatus may be involved in the compartmentalization of GAPDH from cytosol to nuclei. PMID- 11872250 TI - Differential effects of remacemide and desglycinyl-remacemide on epileptiform burst firing in the rat hippocampal slice. AB - Remacemide is a potential anticonvulsant drug with an active metabolite, desglycinyl-remacemide (DGR). Both moieties have been reported to block neuronal Na(+) channels and the N-methyl-D-aspartate (NMDA) subtype of glutamate receptor. The effects of remacemide and DGR on zero Mg(2+)/4-aminopyridine-induced epileptiform discharges were investigated in the rat hippocampal slice preparation and compared with carbamazepine (CBZ), a prototypic Na(+) channel blocker, and AR-R15896AR, a putative NMDA channel blocker. Remacemide (0-100 microM) was without significant effect, while DGR, CBZ and AR-R15896AR all decreased burst frequency in a concentration (0-100 microM) dependent manner. These findings suggest that remacemide is not sufficiently potent at the Na(+) channel or NMDA receptor to attenuate epileptiform activity in this model and that the anticonvulsant effects of the drug may be mediated by DGR. PMID- 11872251 TI - Inhibitory zinc-enriched terminals in the mouse cerebellum: double immunohistochemistry for zinc transporter 3 and glutamate decarboxylase. AB - In the present study, we showed for the first time the presence of inhibitory zinc-enriched neuron terminals in the mouse cerebellar cortex by means of double immunohistochemistry for zinc transporter 3 (ZnT3) and glutamate decarboxylase (GAD). The co-localization of ZnT3 and GAD in the cerebellar cortex was analyzed by confocal microscopy. Strong, punctuate ZnT3-immunoreactivity (Ir) was predominantly distributed in the granule cell layer, while GAD-Ir was seen throughout the cerebellar cortical layers. All of the ZnT3-immunoreactive structures were also immunopositive to GAD, but not vice versa. Based on size and position, these double-labeled elements were axonal terminals of the Golgi and basket cells, in the granule cell and molecular layers, respectively. Observations by electron microscopy revealed that ZnT3-immunoreactive terminals showed typical characteristics of the inhibitory synapses like the following: (1) presynaptic terminals containing flat vesicles; and (2) symmetrical synaptic contacts with dendritic elements. The present results indicate that a zinc containing GABAergic system exists in the mouse cerebellar cortex. PMID- 11872252 TI - Retinoic acid influences Phox2 expression of cardiac ganglionic cells in the developing rat heart. AB - We have studied the expression of the homeodomain transcriptional neuronal regulators Phox2a, Phox2b and the non-neuronal Schwann cell response using the marker S-100 in the differentiating phase of cardiac ganglionic cells in rat embryos following exogenous retinoic acid (RA) treatment of pregnant dams. In control embryos, the expression of Phox2b (E11) preceded that of Phox2a, which, along with the terminal neuronal differentiation marker PGP9.5, was expressed from E12 onwards. Phox2b expression remained unchanged in the differentiated phase of cardiac ganglionic cell development after RA treatment, whereas the population of cells expressing Phox2a, PGP9.5 and S-100 was diminished. These results suggest that RA disrupts the differentiation of cardiac neural crest cells into ganglionic cells destined to contribute to the parasympathetic innervation of the heart, by regulating the expression of Phox2a and Phox2b. PMID- 11872253 TI - GPI1046 prevents dopaminergic dysfunction by activating glutathione system in the mouse striatum. AB - We investigated both the antioxidant activities of GPI1046, a non immunosuppressive derivative of FK506, and the in vivo neuroprotective properties against toxicity of intracerebroventricular 6-hydroxydopamine (6-OHDA) in mice. The 6-OHDA-induced reduction in dopamine and its metabolites in the striatum was significantly normalized by daily administration of GPI1046. Moreover, GPI1046 significantly reduced lipid peroxidation in vivo. Further, GPI1046 significantly increased striatal glutathione (GSH) levels by activating GSH synthesis, although the striatal catalase and superoxide dismutase activities did not change. We conclude that GPI1046 may have neuroprotective effects both in cell cultures and in vivo. PMID- 11872254 TI - Normal development of serotonergic neurons in mice lacking S100B. AB - S100B, a glia-derived calcium binding protein, exhibits strong neurite extension activity in cultured serotonergic neurons. Using S100B-knockout mice, we examined whether this protein possesses in vivo serotonergic trophic activity. The distribution of serotonergic fibers, determined by immunohistochemistry, in the brains of S100B-knockout mice was quite similar to that of wild-type mice. Furthermore, the content of serotonin and its metabolite 5-hydroxyindole-3-acetic acid in knockout mice was also indistinguishable from those of wild-type mice. Our findings argue against the hypothesis that S100B has a crucial role in neurite extension of serotonergic neurons. PMID- 11872256 TI - Differences in white matter architecture between musicians and non-musicians: a diffusion tensor imaging study. AB - Previous studies found structural brain differences between musicians and non musicians. In order to determine possible differences in white matter architecture, diffusion tensor imaging was performed on five adult subjects with musical training since early childhood, and seven adult controls. The musicians displayed significantly greater fractional anisotropy (FA) in the genu of the corpus callosum, while significantly less FA was found in the corona radiata and the internal capsule bilaterally. Further areas also showed significant differences. We hypothesize that these changes are due to the cognitive and motor effects, respectively, of musical training. PMID- 11872255 TI - The neuroimmunophilin GPI-1046 partially protects against 3-acetylpyridine toxicity in the rat. AB - There are no published reports assessing whether the non-immunosuppressant immunophilin ligand GPI-1046, which has putative neurotrophic effects in a variety of models of neurotoxicity, possesses neuroprotective effects in the 3 acetylpyridine (3AP) toxicity model in rats. The purpose of the present study was to assess whether 3AP toxicity is mitigated by administration of GPI-1046. Survival after 3AP challenge and licking performance in surviving rats were used to measure the beneficial effects of GPI-1046. GPI-1046 increased animal survivability and partially ameliorated 3AP-induced performance deficits at the lick task when GPI-1046 was administered concurrently with a peri-lethal dose of 3AP (50 mg/kg). PMID- 11872257 TI - Lithium inhibits amyloid secretion in COS7 cells transfected with amyloid precursor protein C100. AB - To examine the regulation of amyloid secretion in more detail, Abeta sandwich ELISAs with high sensitivity and specificity were developed. Using this technique, we measured Abeta secreted from COS7 cells transiently transfected with APP C100 in the presence of LiCl, a potent glycogen synthase kinase (GSK) 3beta inhibitor. We found that both Abetax-40 and Abetax-42 secretion were reduced by LiCl treatment in a dose-dependent manner. Diminished amyloid secretion was associated with GSK-3beta activity. These results suggest that GSK 3beta might function as a possible mediator for regulating both amyloid deposition and tau pathology in Alzheimer's disease (AD), and that lithium should be re-evaluated as a candidate reagent for preventing AD pathology. PMID- 11872259 TI - Dexterity is not affected by fatigue-induced depression of human motor cortex excitability. AB - Following a fatiguing muscle contraction, motor evoked potentials (MEPs) evoked by transcranial magnetic brain stimulation remain depressed for many minutes, reflecting a reduction in the excitability of the corticospinal projection. No functional significance has been linked to this observation. We postulated that dexterity would be affected when MEPs are depressed. MEPs were recorded from the first dorsal interosseous muscle of 11 healthy subjects in resting muscles before and after a fatiguing maximum voluntary contraction. This induced significant MEP depression in all subjects. No change in dexterity was seen when MEPs were depressed or at any time point throughout the experiment. We conclude that fatigue-induced MEP depression is not associated with a decline in dexterity. PMID- 11872258 TI - Quetiapine attenuates the immobilization stress-induced decrease of brain-derived neurotrophic factor expression in rat hippocampus. AB - Quetiapine is a new atypical antipsychotic drug widely used in the treatment of schizophrenia and other psychotic disorders. This study examined the influence of quetiapine on the decrease of brain-derived neurotrophic factor (BDNF) expression, induced by chronic immobilization stress, in the hippocampus of the rat. Pretreatment with 10 mg/kg of quetiapine markedly attenuated the stress induced decrease in levels of BDNF protein, as determined by Western blot analyses, and the reduction of BDNF immunoreactivity, in hippocampal pyramidal and dentate granular neurons. These results suggest that the chronic administration of quetiapine could be neuroprotective to hippocampal neurons in schizophrenia and this effect may be related to its antipsychotic effect in patients with schizophrenia. PMID- 11872260 TI - Erythropoietin restores glutathione peroxidase activity in 1-methyl-4-phenyl 1,2,5,6-tetrahydropyridine-induced neurotoxicity in C57BL mice and stimulates murine astroglial glutathione peroxidase production in vitro. AB - Recently, we have reported that erythropoietin (Epo) provides neuroprotection in 1-methyl-4-phenyl-1,2,5,6-tetrahydropyridine (MPTP)-induced neurotoxicity in vivo. In the present study, we investigated the effects of single Epo administration on brain antioxidant enzyme (superoxide dismutase (SOD) and glutathion peroxidase (GSHPx)) activities in this model in C57BL/6 mice. We found that MPTP treatment decreased GSHPx activity in both substantia nigra and striatum, and Epo restores nigral GSHPx activity decreased by MPTP. SOD enzyme activity was not significantly changed by MPTP and Epo treatment. Further, Epo stimulated astroglial GSHPx production in neonatal murine astroglial cell culture suggesting that the possible cell source for the stimulation of GSHPx activity by Epo in the MPTP-induced neurotoxicity model are astroglia. In conclusion, modulation of the astroglial antioxidant defense system might be one of the mechanisms by which Epo exerts a beneficial effect in MPTP-induced Parkinsonism. PMID- 11872261 TI - Mental set can modulate response onset in the lower limb muscles to falls in humans. AB - The objective of this study was to investigate the modulation of the ankle muscle electromyographic (EMG) response as a function of mental set. Thirteen young healthy subjects underwent 40 unexpected and self-initiated drops from 30 cm above two separate force-plates. Following unexpected drops, reflex activities were observed in the medial gastrocnemius (MG) and tibialis anterior (TA) at mean latencies (+/- SD) of 83.59 +/- 10.1 and 99.43 +/- 21.82 ms, respectively. Following self-initiated drops, the response latency of the MG was significantly shortened (to 71.98 +/- 10 ms, P<0.05), and the TA was significantly lengthened (to 183.33 +/- 45 ms, P<0.05) when compared with unexpected drops. Such a modulation was associated with a significant reduction of the impact force on landing as compared with unexpected drops (by 17%, P<0.05). Interestingly, a negative correlation was found between the onset of the TA EMG response and the magnitude of the impact force on landing during expected (r= -0.66, P<0.05) but not unexpected drops. PMID- 11872262 TI - Thioredoxin suppresses 1-methyl-4-phenylpyridinium-induced neurotoxicity in rat PC12 cells. AB - Thioredoxin (TRX) is a redox-active protein which plays a cytoprotective role against oxidative stress. Geranylgeranylacetone (GGA), used widely as an anti ulcer drug, has been reported to induce TRX as well as heat shock protein 70 (HSP70) in hepatocytes and other cells. 1-Methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP), causes dopaminergic denervation and Parkinsonism in humans. The 1-methyl-4-phenylpyridinium ion (MPP(+)), an active metabolite of MPTP, induces cell death in a rat pheochromocytoma cell line (PC12 cells). We found that MPP(+) suppresses TRX expression in PC12 cells. Overexpression or administration of TRX attenuates MPP(+)-induced neurotoxicity on PC12 cells. Moreover, GGA induces expression of TRX and HSP70 and attenuates MPP(+)-induced toxicity in PC12 cells. These results indicate that TRX and GGA have a possible potential as new therapeutic agents for Parkinson disease. PMID- 11872263 TI - Behavioral, hormonal and histological stress markers of anxiety-separation in postnatal rats are reduced by prepro-thyrotropin-releasing hormone 178-199. AB - We investigated in the present study whether systemic injections of prepro thyrotropin-releasing-hormone 178-199 (PPTRH 178-199) in postnatal 3-days old rat pups can provide ameliorative effects in a model of anxiety-separation disorder. The pups were individually separated from their mother and placed in a novel environment. PPTRH 178-199-treated animals started exploring the novel environment in a significantly shorter time and elicited significantly less distress vocalizations than control animals. PPTRH 178-199-treated animals also had markedly lower serum adrenocorticotropic hormone and corticosterone compared to control animals. Furthermore, we observed a significant increase in PPTRH 178 199 immunoreactive cell bodies in the hypothalamus of PPTRH 178-199-treated animals compared to controls, suggesting that the peptide crossed the blood-brain barrier. PPTRH 178-199 treatment can help to reduce behavioral and hormonal disturbances associated with anxiety-separation situations. PMID- 11872264 TI - Long-term changes in calbindin D(28K) immunoreactivity in the rat hippocampus after cardiac arrest. AB - Calbindin D(28K) (CB) expression was analyzed in the rat hippocampus following 10 min-cardiac arrest-induced ischemia within a year after reperfusion. In rats examined 3 days after ischemia, CB immunoreactivity disappeared completely from CA1 pyramidal neurons and from most CA2 pyramids. In the stratum granulosum of the dentate gyrus, mossy fibers, and hippocampal interneurons, CB immunoreactivity was preserved, although staining was somewhat paler than that in control rats. A similar pattern of CB immunoreactivity was found in rats sacrificed 14 days and 1 month after cardiac arrest. From the 14th postischemic day, neuronal loss in the stratum pyramidale of CA1 but not in that of CA2 became apparent. The reappearance of CB immunoreactivity in CA1 and CA2 pyramidal neurons was noticed 6 months after ischemia, and the pattern was identical to that observed in animals sacrificed 12 months after the ictus. The prolonged loss and delayed reappearance of CB immunoreactivity in the hippocampus demonstrate that ischemia may induce long-term disturbances of protein expression, which may in turn result in impairment of hippocampal functioning. PMID- 11872265 TI - Neither the density nor function of striatal dopamine transporters were influenced by chronic n-3 polyunsaturated fatty acid deficiency in rodents. AB - We hypothesized that the chronic dietary deficiency of n-3 polyunsaturated fatty acids (n-3 PUFAs) might affect the density and/or function of dopamine transporters (DAT), which have a major role in regulating the synaptic level of dopamine. This hypothesis was tested by investigating DAT in the striatum using three complementary methods in control and deficient rats. The density of DAT was determined by quantitative autoradiography using [(125)I]PE2I, a specific ligand of this transporter. Functional investigations were performed (i) in vitro by measuring [(3)H]dopamine uptake on synaptosomes, and (ii) in vivo using intracerebral microdialysis. The results demonstrated that neither the density nor the function of DAT were influenced by n-3 PUFA deficiency in the striatum. This suggests lower sensitivity to n-3 PUFA deficiency in the striatum than that previously observed in the frontal cortex. PMID- 11872266 TI - Beneficial effects of alpha-lipoic acid plus vitamin E on neurological deficit, reactive gliosis and neuronal remodeling in the penumbra of the ischemic rat brain. AB - During cerebral ischemia-reperfusion, the enhanced production of oxygen-derived free radicals contributes to neuronal death. The antioxidants alpha-lipoic acid and vitamin E have shown synergistic effects against lipid peroxidation by oxidant radicals in several pathological conditions. A thromboembolic stroke model in rats was used to analyze the effects of this mixture under two oral treatments: intensive and prophylactic. Neurological functions, glial reactivity and neuronal remodeling were assessed after experimental infarction. Neurological recovery was only found in the prophylactic group, and both antioxidant schemes produced down-regulation of astrocytic and microglial reactivity, as well as higher neuronal remodeling in the penumbra area, as compared with controls. The beneficial effects of this antioxidant mixture suggest that it may be valuable for the treatment of cerebral ischemia in humans. PMID- 11872267 TI - Water-soluble chitosan inhibits the production of pro-inflammatory cytokine in human astrocytoma cells activated by amyloid beta peptide and interleukin-1beta. AB - A chronic inflammatory response associated with beta-amyloid (Abeta) and interleukin-1beta (IL-1beta) is responsible for the pathology of Alzheimer's disease (AD). Astrocytes are predominant neuroglial cells of the central nervous system and are actively involved in cytokine-mediated events in AD. To investigate the biological effect of water-soluble chitosan (WSC), we examined cytotoxicity, production of pro-inflammatory cytokines and inducible nitric-oxide synthase (iNOS) on human astrocytoma cell line CCF-STTG1 stimulated with IL-1beta and Abeta fragment 25-35 (Abeta[25-35]). In 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyltetrazoliumbromide colorimetric assay, WSC by itself had no effect on cell viability on human astrocytoma cells. The effects of WSC on tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) were evaluated with enzyme linked immunosorbent assay and Western blotting. The production of TNF-alpha and IL-6 was induced by IL-1beta and Abeta[25-35] and synergistically amplified by the co-stimulation of IL-1beta and Abeta[25-35]. The secretion and expression of pro-inflammatory cytokines, TNF-alpha and IL-6, was significantly inhibited by pretreatment with WSC in human astrocytoma cells. The expression of iNOS was induced by IL-1beta and Abeta[25-35] and was partially inhibited by treatment with WSC. We demonstrate the regulatory effects of WSC in human astrocytes for the first time and suggest the anti-inflammatory effect of WSC may reduce and delay AD pathologic events. PMID- 11872268 TI - S-adenosyl-L-methionine prevents 5-HT(1A) receptors up-regulation induced by acute imipramine in the frontal cortex of the rat. AB - S-adenosyl-L-methionine (SAM) has shown efficacy in speeding the onset of the antidepressant effect of imipramine in depressed patients. This effect may be related to their interactions at the serotonin(1A) (5-HT(1A)) receptors. Acute imipramine up-regulated the frontal cortex 5-HT(1A) receptors (B(max), 51.5 +/- 8.4 fmol/mg protein) vs. saline (B(max), 27.5 +/- 5.9 fmol/mg protein), and did not show antidepressant effect. Acute SAM and imipramine+SAM did not modify frontal cortex 5-HT(1A) receptors, and showed antidepressant effects (decrease of the immobility response of 26%, P<0.01; and 47%, P<0.001) vs. saline. All the chronic treatments showed antidepressant effects and up-regulated the hippocampus 5-HT(1A) receptors. SAM prevents the 5-HT(1A) receptor up-regulation induced by acute imipramine in the frontal cortex. This mechanism may contribute to imipramine's antidepressant effect. PMID- 11872269 TI - A brain nitric oxide synthase study in the rat: production of a nitroso-compound NA and absence of nitric oxide synthesis. AB - The products of brain NO-synthase (NOS) were studied by different analytical techniques with the same incubation conditions. Voltammetric techniques used a micro cell containing NOS and its substrate (10 mM arginine). Using porphyrin microelectrodes with differential pulse amperometry nitric oxide (NO) was not detected when nafion membrane was present (less than 0.3 muM). Nitrite was detected with the same microelectrode without membrane (0.42 mM). Differential pulse voltammetry (DPV) with micro carbon electrode detected a nitroso-compound (NA) in reduction (1 mM) and not NO. In oxidation the observed DPV peak was due to nitrite (0.43 mM). Citrulline was detected by high performance liquid chromatography (0.51 mM). Using Diels Alder reaction in NOS preparation a NA cycloadduct was observed by capillary electrophoresis (0.2 mM) and mass spectrometry (0.22 mM). Diels Alder reaction is the reaction of the identification of the nitroso group. NA-cycloadduct degradation by retro Diels Alder reaction gave equimolar concentrations of citrulline and nitrite without NO production. These observations lead us to affirm that NOS synthesizes NA. PMID- 11872270 TI - Baclofen inhibits ANP-mediated cyclic GMP synthesis in the rat cervical spinal cord. AB - The neurotransmitter gamma-aminobuteric acid (GABA) is believed to have a controlling action on spinal locomotor networks. In spasticity, spinal locomotor networks are thought to play a role. A well known drug in the treatment of spasticity is the GABA(B) agonist Baclofen. We report an inhibitory effect of Baclofen on the ANP-mediated cGMP synthesis in the superficial dorsal horn (laminae I-III) of the rat cervical spinal cord. This inhibitory effect of Baclofen could not be detected after incubation with the NO donor SNP. The clinical effect of Baclofen on the reduction of spasticity might be explained by an enhancement of GABAergic inhibition of ANP mediated cGMP concentration in the spinal cord dorsal horn, thus reducing afferent input. PMID- 11872271 TI - Elimination of phenolsulfonphthalein from the cerebrospinal fluid via capillaries in central nervous system in cats by active transport. AB - It was recently proposed that organic anions, such as cerebral acidic metabolites and phenolsulfonphthalein (PSP), are eliminated from cerebrospinal fluid (CSF) by diffusion into the central nervous system (CNS) and further by active transport into capillaries. To test this hypothesis, PSP was injected into cisternal CSF and its distribution into various parts of the CNS was measured 1 and 3 h later in control cats and those pretreated with probenecid, which blocks active transport of organic anions into capillaries. PSP in tissue shows an intensive pink color when exposed to 1 N NaOH. Planimetric analysis of color pictures of coronal CNS slices showed that at the first hour, diffusion and distribution of PSP into the CNS in both groups of animals was similar, while at the third hour, a great reduction of PSP distribution in the CNS in control and only a slight reduction in probenecid pretreated cats was observed. The results support the hypothesis that active transport across the capillary wall in the CNS is the main avenue for elimination of cerebral acidic metabolites from both CSF and CNS and in such a way that central homeostasis is maintained. PMID- 11872272 TI - Gastric surgical adjuvant radiotherapy consensus report: rationale and treatment implementation. AB - PURPOSE: Radiation therapy has recently emerged as a pivotal modality in the management of completely resected, high-risk gastric cancer. The recently published results of the Intergroup 0116 Gastric Surgical Adjuvant Trial randomized high-risk (T3,4 and/or node positive), completely resected gastric or gastroesophageal adenocarcinomas to receive either observation alone or radiochemotherapy after complete resection. Radiochemotherapy produced significant improvements in relapse-free (p < 0.0001) and overall survival (p = 0.01). Radiation oncologists must now clearly comprehend the principles governing the rationale supporting this therapy to apply it to those afflicted with this disease. This paper represents a consensus report reviewing data supporting radiotherapy, important clinical and anatomic issues related to radiotherapy, and details of the practical application of radiation therapy to commonly occurring clinical presentations. Supportive therapy during and after radiochemotherapy is also discussed. PMID- 11872273 TI - Apoptosis as a cellular predictor for histopathologic response to neoadjuvant radiochemotherapy in patients with rectal cancer. AB - BACKGROUND: Tumor shrinkage by preoperative radiochemotherapy (RCT) can markedly improve surgery in locally advanced (T4) rectal cancer with clear resection margins and may enable sphincter preservation in low-lying tumors. However, tumor response varies considerably, even among tumors treated according to the same protocol. If one is able to identify patients with highly radio-responsive tumors at the time of diagnosis, a selective and individualized policy of preoperative RCT might be pursued. METHODS: The apoptotic index (AI), Ki-67, p53, and bcl-2 were evaluated by immunohistochemistry on pretreatment biopsies from 44 patients treated uniformly according to a prospective neoadjuvant RCT protocol (CAO/AIO/ARO-94). Treatment response was assessed histopathologically in the resected surgical specimen, using a five-point grading system. Expression of each marker was correlated with tumor response and relapse-free survival after curative surgery. RESULTS: Tumors with complete (n = 3) or good (n = 28) response to RCT showed significantly higher pretreatment levels of apoptosis (mean AI: 2.06%) than tumors with moderate (n = 7), minimal (n = 5), or no regression (n = 1) from RCT (AI: 1.44%, p = 0.003). The AI was significantly related to Ki-67 (p = 0.05), but not to p53 and bcl-2 status. Tumor regression and AI best predicted relapse-free survival after combined modality treatment and curative surgery. CONCLUSION: Spontaneous apoptosis in rectal cancer may serve as an important predictor of tumor regression from RCT in rectal cancer and as a significant prognosticator of relapse-free survival. Thus, this molecular marker may finally help to tailor therapy with regard to (neo-) adjuvant treatment of rectal cancer. PMID- 11872274 TI - Accelerated radiochemotherapy in pancreatic cancer is not necessarily related to a pathologic pancreatic function decline in the early period. AB - PURPOSE: To evaluate the functional effects of ionizing radiation in patients with unresectable pancreatic cancer in the early period after accelerated radiochemotherapy (ART). METHODS AND MATERIALS: To analyze the exocrine component, the amino acid consumption test and fecal elastase 1 were performed in 13 patients immediately before and 4-8 weeks after ART. Pancreatic duct morphology was evaluated before therapy. Weight loss and clinical steatorrhea were recorded. Endocrine parameters were examined according to standardized criteria. RESULTS: The relative change of the amino acid consumption test results and the median elastase concentration was 41.2% and 56.4%, respectively. Five patients still had normal test results after ART and 5 patients developed pathologic values. The median relative weight loss of the total body weight was 7.7% +/- 4.5%. No steatorrhea occurred. Of the 5 patients with normal values, 3 had a mean organ dose of <40 Gy. Of the 5 patients with pathologic values, 4 had a mean organ dose of >41 Gy. The endocrine function measurements remained unchanged. CONCLUSION: Although a nominal reduction of exocrine function parameters occurred in most patients, ART was not necessarily related to a pathologic level in the early period. Diabetes was not established. The functional impairment that was existent in the patient population presumably contributed to the weight loss. Pancreatic enzyme preparations may also play a role in maintaining an anabolic state during and after radiochemotherapy. PMID- 11872275 TI - Palliation of advanced/recurrent esophageal carcinoma with high-dose-rate brachytherapy. AB - PURPOSE: The aim was to assess the improvement in swallowing status, complication rate, and overall survival. PATIENTS AND METHODS: Fifty-eight patients with advanced/recurrent esophageal carcinoma were treated for palliation with high dose-rate intraluminal brachytherapy (HDR-ILRT) with a remote afterloading microSelectron unit (192Ir source) with or without external radiation from November 1994 to May 2000 at the Tata Memorial Hospital. The mean age was 64 years. The mid-third of the esophagus was involved in 38 patients (66%). The group was comprised of 37 previously untreated patients (29 with < or = 50% Karnofsky performance status and old age, 4 with metastatic disease, and 4 with second primary esophageal lesions) and 21 patients with post-treatment recurrent tumors. Thirty-eight patients (65%) received intraluminal brachytherapy alone, whereas the remaining 20 patients (35%) received a combination of external and intraluminal radiation therapy. All patients received 2 fractions of HDR-ILRT 1 week apart with 600 cGy per fraction at 1 cm off axis. RESULTS: Overall improvement in swallowing status was seen in 22 patients (48%), and 24 (41%) maintained pretreatment swallowing status. Median dysphagia-free survival was 10 months. Overall complication rates were 30%, with stricture seen in 9 patients (15%), ulceration in 6 (10%), and tracheo-esophageal fistula in 3 patients (5%). Complication rates were higher in the post-treatment group (38%) than in the previously untreated group (27%) (p = 0.29). The median overall survival for the entire group was 7 months. Median survival was better, although not significantly, for the previously untreated cohort: 7.8 months vs. 6 months for the post-treatment group (p = 0.77). CONCLUSION: HDR-ILRT brachytherapy achieves good palliation with acceptable complications in advanced/recurrent esophageal carcinoma. PMID- 11872276 TI - Randomized trial on the efficacy of radiotherapy for cerebral low-grade glioma in the adult: European Organization for Research and Treatment of Cancer Study 22845 with the Medical Research Council study BRO4: an interim analysis. AB - PURPOSE: There is no consensus on the treatment strategy for adult patients with cerebral low-grade glioma. The diagnosis and primary treatment are usually undertaken by surgery. Some investigators doubt the efficacy of postoperative radiotherapy (RT), whereas others advise routine postoperative RT. We report the primary results of a multicenter randomized trial on this controversy. METHODS AND MATERIALS: From 24 European centers, 311 adult patients with low-grade glioma were randomized centrally after surgery from March 1986 through September 1997, between the two arms of the trial. The irradiated group received 54 Gy in 6 weeks. The other patients did not receive any treatment after surgery until the tumor showed progression, defined as clinical-neurologic deterioration and evidence of progressive tumor on imaging. RESULTS: Of 290 eligible and assessable patients (93%), the irradiated group showed a significant (log-rank p = 0.02) improvement in time to progression but not in overall survival, with a median follow-up of 5 years. The 5-year estimate was, respectively, 63% vs. 66% (overall survival) and 44% vs. 37% (time to progression) for the treated and control arms. Different treatment modalities, including RT, were undertaken for the 85 controls when a progressive tumor was noted. CONCLUSION: Early postoperative conventional RT such as that used for this protocol appears to improve the time to progression or progression-free survival, but not overall survival, for patients with low grade glioma. PMID- 11872277 TI - Preliminary results from a Phase II trail of conformal radiation therapy for pediatric patients with localised low-grade astrocytoma and ependymoma. AB - PURPOSE: To estimate the local control and patterns of failure for pediatric patients with low-grade astroglial tumors (LGA) and ependymoma (EP) treated with three-dimensional conformal radiation therapy (CRT) using an anatomically defined clinical target volume (CTV). METHODS AND MATERIALS: From an ongoing, prospective Phase II trial initiated in July 1997, 102 pediatric patients with LGA (n = 38) and EP (n = 64) have been treated with CRT using an anatomically defined CTV extending 1.0 cm beyond the gross tumor volume and a 0.5-cm margin (planning target volume) extending outside of the CTV. The prescribed dose was 54 Gy (LGA) and 59.4 Gy (EP). RESULTS: Patients with EP have been followed for a median of 17 months (range 3--43 months), and six failures have occurred. Patients with LGA have been followed for a median of 17 months (3--44 months), and four failures have occurred. Three-dimensional magnetic resonance (MR) studies performed to document treatment failure were registered with the MR and computed tomography (CT) data used in the treatment planning process. Failure occurred within the CTV for 5 patients with EP, including 3 with concurrent subarachnoid dissemination. One patient with EP developed metastatic disease with no evidence of local failure. Three patients with LGA failed within the CTV and one failed immediately outside of the CTV. CONCLUSIONS: Treatment of an anatomically defined CTV, encompassing 1.0 cm of non-involved brain beyond the margin of resection or neuroimaging-defined tumor, appears to be safe for pediatric patients with LGA and EP based on these preliminary data. Normal tissue sparing through the use of advanced radiation therapy treatment planning and delivery techniques should be beneficial to pediatric patients if the rate and patterns of failure are similar to conventional techniques and toxicity reduction can be objectively documented. PMID- 11872278 TI - Risk of symptomatic brain tumor recurrence and neurologic deficit after radiosurgery alone in patients with newly diagnosed brain metastases: results and implications. AB - PURPOSE: A single-institution experience using primary stereotactic radiosurgery (SRS) alone in the management of newly diagnosed brain metastases was analyzed to identify the risk of symptomatic brain tumor recurrence (BTR) and neurologic deficit associated with such a treatment strategy. METHODS AND MATERIALS: Thirty six patients were treated for newly diagnosed single/multiple brain metastases using SRS alone followed by planned observation. SRS minimum tumor dose ranged from 8 to 25 Gy (median: 20 Gy). Factors evaluated in analysis of treatment outcome included number of metastases, site of metastasis, primary tumor site, histology, extent of intracranial and extracranial disease, and interval to diagnosis of brain metastasis. RESULTS: Median and 1-year survival for the entire group was 9 months and 36%, respectively. BTR anywhere in the brain occurred in 47% (17/36) of patients. Forty-seven percent of BTR (8/17) recurred at the site of original metastasis; 35% (6/17) recurred at both original [corrected] and distant sites in the brain, and 18% (3/17) recurred at distant only [corrected] brain sites. Seventy-one percent (12/17) of the patients were symptomatic at the time of recurrence, and 59% (10/17) had an associated neurologic deficit. Multivariate analysis found that only the extent of disease was a predictor of BTR. Patients who had disease limited to the brain only had a BTR rate of 80% (8/10) vs. 35% (9/26) who had disease involving the brain, primary site, and/or other extracranial metastatic sites (p = 0.03). CONCLUSIONS: Use of primary SRS alone in this setting is associated with an increasingly significant risk of BTR with increasing survival time. In addition, the majority of such recurrences are symptomatic and associated with a neurologic deficit, a finding not analyzed in recently reported experiences withholding whole brain radiation therapy as part of the primary treatment of brain metastasis. PMID- 11872279 TI - The impact of (18)FDG-PET on target and critical organs in CT-based treatment planning of patients with poorly defined non-small-cell lung carcinoma: a prospective study. AB - PURPOSE: To prospectively study the impact of coregistering (18)F-fluoro-deoxy-2 glucose hybrid positron emission tomographic (FDG-PET) images with CT images on the planning target volume (PTV), target coverage, and critical organ dose in radiation therapy planning of non-small-cell lung carcinoma. METHODS AND MATERIALS: Thirty patients with poorly defined tumors on CT, referred for radical radiation therapy, underwent both FDG-PET and CT simulation procedures on the same day, in radiation treatment position. Image sets were coregistered using external fiducial markers. Three radiation oncologists independently defined the gross tumor volumes, using first CT data alone and then coregistered CT and FDG PET data. Standard margins were applied to each gross tumor volume to generate a PTV, and standardized treatment plans were designed and calculated for each PTV. Dose-volume histograms were used to evaluate the relative effect of FDG information on target coverage and on normal tissue dose. RESULTS: In 7 of 30 (23%) cases, FDG-PET information changed management strategy from radical to palliative. In 5 of the remaining 23 (22%) cases, new FDG-avid nodes were found within 5 cm of the primary tumor and were included in the PTV. The PTV defined using coregistered CT and FDG-PET would have been poorly covered by the CT-based treatment plan in 17--29% of cases, depending on the physician, implying a geographic miss had only CT information been available. The effect of FDG-PET on target definition varied with the physician, leading to a reduction in PTV in 24 70% of cases and an increase in 30-76% of cases. The relative change in PTV ranged from 0.40 to 1.86. On average, FDG-PET information led to a reduction in spinal cord dose but not in total lung dose, although large differences in dose to the lung were seen for a few individuals. CONCLUSION: The coregistration of planning CT and FDG-PET images made significant alterations to patient management and to the PTV. Ultimately, changes to the PTV resulted in changes to the radiation treatment plans for the majority of cases. Where possible, we would recommend that FDG-PET data be integrated into treatment planning of non-small cell lung carcinoma, particularly for three-dimensional conformal techniques. PMID- 11872280 TI - Early mortality after radical radiotherapy for non-small-cell lung cancer: comparison of PET-staged and conventionally staged cohorts treated at a large tertiary referral center. AB - PURPOSE: At our center, approximately 30% of radical radiotherapy (RRT) candidates become ineligible for RRT for non-small-cell lung cancer (NSCLC) after positron emission tomography (PET). We hypothesized that early cancer death rates would be lower in patients receiving RRT after PET staging compared with conventionally staged patients. METHODS AND MATERIALS: Two prospective cohorts were compared. Cohort 1 consisted of all participants in an Australian randomized trial from our center given 60 Gy conventionally fractionated RRT with or without concurrent carboplatin from 1989 to 1995. Eligible patients had Stage I--III, Eastern Cooperative Oncology Group status 0 or 1, <10% weight loss, and had not undergone PET. Cohort 2 included all RRT candidates between November 1996 and April 1999 who received RRT after PET staging and fulfilled the above criteria for stage, Eastern Cooperative Oncology Group status, and weight loss. RESULTS: Eighty and 77 eligible patients comprised the PET and non-PET groups, respectively. The PET-selected patients had significantly less weight loss; 73% and 49% of the PET and non-PET patients, respectively, received chemotherapy. The median survival was 31 months for PET patients and 16 months for non-PET patients. Mortality from NSCLC and other causes in the first year was 17% and 8% for PET patients and 32% and 4% for non-PET patients, respectively. The hazard ratio for NSCLC mortality for PET vs. non-PET patients was 0.49 (p = 0.0016) on unifactorial analysis and was 0.55 (p = 0.0075) after adjusting for chemotherapy, which significantly improved survival. CONCLUSION: Patients selected for RRT after PET have lower early cancer mortality than those selected using conventional imaging. PMID- 11872281 TI - Impact of institutional experience on survival outcome of patients undergoing combined chemoradiation therapy for inoperable non-small-cell lung cancer. AB - PURPOSE: Clinical experience of both physicians and institutions has been shown to significantly influence the outcome of patients. We conducted this retrospective cohort study to examine its impact on the outcome of patients undergoing combined chemoradiation therapy for the treatment of locally advanced inoperable non-small-cell lung cancer. METHODS AND MATERIALS: We compared the clinical data from 239 patients who were enrolled in two consecutive Radiation Therapy Oncology Group (RTOG) trials (RTOG 91-06, RTOG 92-04) according to the number of patients enrolled from each institution in either trial alone or the two trials combined. RESULTS: Overall, patients treated at the institutions that enrolled > or = 5 patients survived longer than those treated at the institutions that enrolled <5 patients (median survival 20.5 vs. 13.4 months, p = 0.0006) with a more than doubling of the 2- and 3-year survival rates (45% and 31% vs. 20% and 13%, respectively). Multivariate analyses confirmed that the number of patients enrolled from each institution was an important prognostic factor for the entire group (p = 0.001) and also for RTOG 91-06 (p = 0.05) and RTOG 92-04 (p = 0.004) when the data were analyzed separately. CONCLUSION: Institutional experience has a significant impact on the survival outcome of patients undergoing combined chemoradiation therapy for inoperable non-small cell lung cancer. PMID- 11872283 TI - Dose, volume, and tumor control prediction in primary radiotherapy of non-small cell lung cancer. AB - BACKGROUND: To evaluate the influence of total dose and tumor volume on local control and survival in primary radiotherapy of non-small-cell lung cancer (NSCLC). METHODS AND MATERIALS: We retrospectively analyzed the clinical course and CT-derived pre- and post-therapeutic tumor volume data of 135 patients with NSCLC undergoing primary radiotherapy at our department between 1989 and 1996. Among these, a total of 192 spatially separated tumor volumes (135 primary tumors, 1 additional intrapulmonary tumor, and 56 involved lymph nodes) were available for analysis. In all patients, treatment was planned using CT-based three-dimensional treatment planning. The dose to each tumor volume was derived from the individual dose plans. Mean total dose was 59.9 Gy (range: 30-80 Gy). All but 3 patients were followed until death. For local control analysis, each tumor was analyzed separately, and its remission status was determined in serial follow-up CT scans. A total of 784 CT scans were analyzed. Actuarial local control analysis was performed for the 192 separated tumor volumes, and survival analysis was performed for the 135 patients. Tumor control probability was calculated using a Poisson statistical model. RESULTS: Overall 1- and 2-year local control rate was 50% and 37%, respectively. The 2-year local control rate for tumors <50 ccm, 50-200 ccm, and >200 ccm was 51%, 22%, and 10%, respectively (p = 0.02). The 2-year local control rate for dose levels < or = 60 Gy and >60 Gy was 28% and 43% (p < 0.001). For the subgroup of 147 tumors smaller than 100 ccm, the local control rate increased up to 70% (1 year) and 51% (2 years) with doses of more than 60 Gy. For tumors larger than 100 ccm, no dose effect was seen. Only 2 of 45 tumors >100 ccm were controlled more than 2 years. Multivariate analysis revealed tumor volume, total dose, histopathologic type, and grading as significant and independent prognostic factors for local control. The number of delay days by split course (if used) and application of chemotherapy was not found to influence local control. Overall 1- and 2-year survival rate was 42% and 13%. Total radiation dose, chemotherapy, and T and N stage---but not tumor volume --were found to be independent and significant prognostic factors for survival in multivariate analysis. CONCLUSION: Tumor volume is an important predictor of local control in NSCLC. We found a clear dose effect for local control and survival in NSCLC. Long-term local control for a significant proportion of patients seems possible for small tumors only (<100 ccm, i.e., maximum diameter 6 cm) with doses of 70 Gy and more. Tumors of > or = 100 ccm are unlikely to be controlled long term by conventional doses up to 70 Gy. These results support dose escalation in patients with NSCLC. PMID- 11872282 TI - A quality-adjusted reanalysis of a Phase III trial comparing once-daily thoracic radiation vs. twice-daily thoracic radiation in patients with limited-stage small cell lung cancer(1). AB - PURPOSE: We undertook an analysis of quality-adjusted survival using the Q-TWiST (Quality Time Without Symptoms or Toxicity) methodology and developed a new graphic representation called a quality-adjusted life-years plot, which presents a complete and concise Q-TWiST analysis on a single plot. METHODS AND MATERIALS: The Q-TWiST plot incorporates the time without symptoms or toxicity and several combinations of utility coefficients for toxicity and relapse days into the same plot. In addition, the plot includes threshold lines, to judge whether a particular combination of utility coefficients reaches a significance level. RESULTS: The differential in toxicity incidence and severity between the two thoracic radiation treatment arms was inconsequential. Sensitivity analyses were run using Q-TWiST plots. For all combinations of the various toxicity definitions and utility coefficients, the median Q-TWiST was greater for the once-daily thoracic radiation treatment arm than for the twice-daily treatment arm, without achieved significance. CONCLUSION: This work refines the results previously reported for this Phase III clinical trial in patients with limited-stage small cell cancer, and there was no significant difference in survival after adjusting for toxicity and progression. Furthermore, the new methods developed for this trial allow for a more detailed and parsimonious presentation of survival and toxicity data for all oncology clinical trials. PMID- 11872284 TI - Reirradiation for locally recurrent lung cancer previously treated with radiation therapy. AB - PURPOSE: Local recurrence of lung cancer after previous external beam irradiation poses some problems for subsequent management. We retrospectively reviewed our series of patients with local recurrence of lung cancer to evaluate the efficacy and safety of reirradiation. PATIENTS AND METHODS: Between 1979 and 2000, 34 patients with local recurrence of lung cancer were retreated with external radiation. There were 29 males and 5 females ranging in age from 38 to 85 years (median: 69 years). At the time of reirradiation, the clinical stage was I or II in 2 patients, IIIa in 5 patients, IIIb in 14 patients, and IV in 13 patients. Reirradiation was performed in 18 patients with the aim of achieving a cure or prolongation of survival (radical treatment), while 16 patients were treated for improvement of their symptoms (symptomatic treatment). RESULTS: The median interval between the initial radiation therapy and reirradiation was 23 months, with a range of 5 to 87 months. The dose of initial irradiation delivered to the tumor ranged from 30 to 80 Gy (median: 60 Gy) in 1.5--2.0-Gy fractions per day. During reirradiation, it ranged from 10 to 70 Gy (median: 50 Gy) in 1.8--3.0-Gy fractions per day. The cumulative dose delivered to the tumor by treatments of both initial and second irradiation ranged from 56.5 to 150 Gy (median: 110 Gy). A response was observed in 14 out of 18 patients given radical treatment (complete response, 6; partial response, 8). Twelve of the 16 patients (75%) given symptomatic treatment also showed a symptomatic benefit. The overall survival rate after reirradiation was 43% at 1 year and 27% at 2 years, with a median survival time of 8 months. The median survival time after radical treatment was 15 months, with a range of 3 to 58 months, whereas that after symptomatic treatment was 3 months, with a range of 1 to 14 months. Six long-term survivors lived for more than 20 months. Reirradiation-induced toxicity included symptomatic radiation pneumonitis in 19 patients and symptomatic radiation esophagitis in 6 patients. These toxicities were not fatal, and radiation myelopathy was not caused by reirradiation. CONCLUSION: Based on this study, external beam reirradiation can achieve satisfactory results for local recurrence of lung cancer provided that attention is paid to the possible hazards. PMID- 11872285 TI - Low HER2/neu gene expression is associated with pathological response to concurrent paclitaxel and radiation therapy in locally advanced breast cancer. AB - PURPOSE: The objective of this study was twofold: first, to identify patients with locally advanced breast cancer (LABC) who will achieve a pathological response to a preoperative regimen of concurrent paclitaxel and radiation; and second, to explore associations between molecular markers from the original tumors and pathological response. METHODS AND MATERIALS: Patients with previously untreated LABC were eligible to receive a regimen of preoperative concurrent paclitaxel, 30 mg/m(2) twice a week for a total of 8 weeks, and radiation delivered Weeks 2--6, 45 Gy at 1.8 Gy per fraction to the breast, ipsilateral axilla, and supraclavicular nodes. At mastectomy, pathologic findings were classified as pathological complete response (pCR) = no residual invasive cells in the breast and axillary contents; pathological partial response (pPR) = presence of < or = 10 microscopic foci of invasive cells; no pathological response (pNR) = pathological persistence of tumor. For each patient, pretreatment breast cancer biopsies were prospectively analyzed by immunohistochemistry (IHC) for estrogen and progesterone (ER/PR) hormonal receptors, HER2/neu and p53 overexpression. Estrogen receptor (ER), HER2/neu, metablastin, beta-tubulin III and IV, microtubule-associated protein-4 (MAP-4), bcl-2, bax, and cyclooxygenase-2 (COX-2) gene expression were measured using real time quantitative polymerase chain reaction (PCR). RESULTS: A total of 36 patients had pretreatment biopsies and were evaluable for the analysis of the association of molecular markers with pathological response. Pathological response in the mastectomy specimen was achieved in 12 of these 36 patients (33%). Only HER2/neu and ER gene expression were found to be significantly associated with the extent of pathological response to the regimen, i.e., tumors with low HER2/neu gene expression and negative estrogen receptors were more likely to respond to the tested regimen (p = 0.009 and p = 0.006, respectively). Conversely, p53 protein expression measured by IHC did not appear to be associated with pathological response (p = 0.67). CONCLUSION: Further studies in LABC should assess whether patient selection for treatment based on the original tumor molecular characteristics could affect their chance to achieve a pathological response. PMID- 11872286 TI - Second nonbreast malignancies after conservative surgery and radiation therapy for early-stage breast cancer. AB - PURPOSE: Breast cancer patients treated with conservative surgery and radiation therapy are at risk of developing second nonbreast malignancies (SNBMs). The purpose of this study was to determine the incidence of all SNBMs and SNBMs by specific location among long-term survivors and to compare the risk of these events to the age-specific incidence of malignancies as first cancers in the Surveillance Epidemiology and End-Results Program (SEER) population. METHODS AND MATERIALS: We analyzed the likelihood of SNBM development for 1884 patients with clinical Stage I or II breast cancer treated with gross excision and > or = 60 Gy (median 63) to the breast between 1970 and 1987. Fifty-seven percent received supraclavicular/axillary radiation (median dose 45 Gy, range 20-60) and 28% received systemic therapy. The median age at diagnosis was 52 years. The median clinical tumor size was 2 cm. Patients were considered at risk of an SNBM until the development of the first of distant metastases or contralateral breast cancer or death or, if alive and disease-free, until the last follow-up visit. The expected numbers of cancers were obtained from the SEER database, using the age specific incidence for white women within 5-year age groups and 5-year calendar intervals. The median time at risk for an SNBM was 10.9 years (range 0.2-27.9). RESULTS: By 8 years of follow-up, 432 patients (23%) had developed distant metastases, 295 patients (16%) a local/regional recurrence, and 159 (8%) a contralateral primary. Of the 1884 patients in our cohort, 147 (8%) developed an SNBM compared with the 127.7 expected from SEER. This corresponds to an absolute excess of 1% of the study population and a relative increase of 15% greater than that expected from SEER (p = 0.05). Within the first 5 years, the observed and expected rates of SNBMs were identical (47 vs. 46.9). After 5 years, 24% more SNBMs were observed than expected (100 vs. 80.8, p = 0.02). Among patients <50 years old at breast cancer diagnosis, 43% more observed SNBMs occurred than expected (40 vs. 28, p = 0.02). For patients > or = 50 years, 7% more SNBMs were observed than expected (107 vs. 99.7, p = 0.25). Lung SNBMs were observed in 33 women, 52% more than the 21.67 predicted by SEER (p = 0.01). Most of the lung SNBMs occurred >5 years after treatment (n = 23) and in women who were >50 years at the time of their breast cancer diagnosis (n = 27). The observed incidence of ovarian cancer was significantly greater than expected among patients <50 years (7 vs. 1.96, p = 0.004) but was not different than expected for patients > or = 50 years (5 vs. 5.3, p = 0.61). Among the 7 sarcomas, 3 developed in the radiation field. CONCLUSIONS: SNBMs occur in a substantial minority (8%) of patients treated with conservative surgery and radiotherapy. However, the absolute excess risk compared with the general population is very small (1%). This excess risk is only evident after 5 years. In particular, a slightly increased incidence of lung SNBMs and a somewhat larger increase in ovarian cancer among younger patients was found. Our data suggest that preventive strategies to reduce the incidence of certain cancers (e.g., smoking cessation and prophylactic oophorectomy) and/or continued monitoring for SNBMs to increase the likelihood of early detection and treatment may be prudent in this population. PMID- 11872287 TI - DNA ploidy analysis performed prospectively using fresh tumor samples in early glottic carcinoma treated with radiotherapy. AB - PURPOSE: The correlation between the DNA content, determined using fresh tumor tissues, and the macroscopic presentation of the lesion was studied prospectively in patients with T1 glottic carcinoma. METHODS AND MATERIALS: DNA flow cytometry and fiber-optic endoscopic examination were performed for 30 previously untreated patients with T1 glottic carcinoma. The patients received radical radiotherapy at Aichi Cancer Center Hospital. RESULTS: In regard to the type of lesion, 4 (80%) were aneuploid, and 1 (20%) was diploid for the invasive type. There was a tendency to show an invasive appearance in aneuploid tumors. With respect to clinical outcome, there were 3 (43%) local recurrences among the aneuploid tumors that invaded the entire length of one vocal cord, 0 (0%) for medium-sized lesions, and 1 (17%) for small lesions. Aneuploid tumors showed a high correlation between lesion size and local control. CONCLUSIONS: The correlation was not strong enough to conclude that DNA content can replace the macroscopic presentation of the lesion. However, the combination of DNA content and tumor size may help predict radiation sensitivity. PMID- 11872288 TI - Post-RT CT results as a predictive model for the necessity of planned post-RT neck dissection in patients with cervical metastatic disease from squamous cell carcinoma. AB - PURPOSE: To establish whether the extent of neck disease on postradiation therapy (RT) computed tomography (CT) can predict the likelihood of positive neck nodes and, thereby, the necessity of planned post-RT neck dissection. METHODS AND MATERIALS: Ninety-five patients who underwent post-RT neck dissection within 2 months for squamous cell carcinoma of the head and neck were eligible. Of the 95 patients, 37 (32.7%) of 113 hemineck specimens were pathologically positive. On post-RT CT imaging studies, the number and size of lymph nodes >1 cm were recorded. Internal focal defects and the likelihood of extracapsular spread were graded. RESULTS: If lymph nodes on post-RT CT were < or = 15 mm, free of significant internal focal low-attenuation or calcification, and without imaging evidence of extracapsular spread, the surgical hemineck specimen was positive in 1 (3.4%) of the 29 hemineck specimens. A focal low-attenuation defect (p = 0.0078) and evidence of extracapsular spread (p = 0.0721) seen in the residual nodal mass on CT were independent predictors of a positive surgical specimen by multivariate analysis. CONCLUSION: CT findings on post-RT neck studies can help predict the likelihood of residual disease and, thereby, the necessity of planned post-RT neck dissection. PMID- 11872289 TI - Magnetic resonance spectroscopic imaging-guided brachytherapy for localized prostate cancer. AB - PURPOSE: Prostate brachytherapy (PB) entails the placement of radioactive sources throughout the entire prostate gland to treat localized cancer. Typically, the target volume in PB encompasses the entire prostate gland because of the inability to localize the cancer and the multifocal nature of this malignancy. However, because of the unique biochemical nature of the prostate gland, recent advances in magnetic resonance spectroscopic imaging (MRSI) of the prostate have allowed precise delineation of the cancer location within the prostate gland. This report reveals our initial experience of MRSI-guided PB. METHODS: A MRSI study was obtained in 15 localized prostate cancer patients before their scheduled PB. The results of this study were used to internally map 7 x 7 x 9-mm volumes of prostate tissue to assign cancerous areas a higher dose of radiation. Such tumor-bearing areas had a low citrate/(choline+creatine) ratio consistent with cancer. On the basis of the anatomic MRI and MRSI correlation, three dimensional coordinates were assigned to the locations of MRSI-defined cancer. The entire target volume was treated to a standard prescription dose using I-125 or Pd-103. Abnormal citrate regions, termed the biologic tumor volume, were prescribed a dose of 130% of the target volume dose to dose escalate in the abnormal citrate regions while respecting the normal radiation tolerances of the surrounding areas. Three-dimensional treatment planning was used to perform the implant. RESULTS: Of the 15 prostate cancer patients evaluated, all had successful three-dimensional MRSI acquisition before their scheduled PB procedure. In 14 of the 15 patients planned with MRSI, the data were successfully incorporated into their treatment planning and were used to increase the radiation dose prescription to 130% in the MRSI-defined volumes. In 1 patient, MRSI revealed significant background artifact that made a focal boost impractical. Postimplant dosimetry confirmed a median V100 of 95% (range 72% 100%) in the 15 evaluated patients for the prescription dose. Furthermore, the median BTV100 for the abnormal citrate region was 90% (range 80-100%) as determined by postimplant dosimetry. Urethral and rectal dose-volume histograms were within normal limits. Morbidity was comparable with that for conventionally treated patients. CONCLUSION: MRSI offers a promising new approach for the delivery of ionizing radiation in PB. Although this series was small and with a short follow-up, MRSI-guided implants are feasible and warrant further investigation as a means of improving the therapeutic ratio in PB [corrected]. PMID- 11872290 TI - Normalization of serum testosterone levels in patients treated with neoadjuvant hormonal therapy and three-dimensional conformal radiotherapy for prostate cancer. AB - PURPOSE: To determine the expected time to serum testosterone normalization after short-course neoadjuvant androgen deprivation therapy (NAAD) and three dimensional conformal radiotherapy for patients with localized prostate cancer and to identify pretreatment predictors that correlated with the time to testosterone normalization. METHODS: Between 1993 and 1999, 88 patients with localized prostate cancer, treated with NAAD and external beam radiotherapy, were prospectively monitored after treatment with sequential testosterone levels. NAAD was administered before and during the entire course of radiotherapy and discontinued at the end of treatment. The median duration of NAAD was 6 months. The actuarial rate of serum testosterone normalization from the end of treatment was evaluated, and the presence or absence of androgen deprivation-related symptoms was correlated with serum testosterone levels. Symptoms assessed included weight gain, loss of libido, breast tenderness, breast enlargement, hot flashes, and fatigue. RESULTS: Serum testosterone levels returned to the normal range in 57 (65%) of the 88 patients and failed to normalize in 31 patients (35%). The median time to normalization was 18.3 months. The actuarial rate of normalization at 3, 6, 12, and 24 months was 10%, 26%, 38%, and 59%, respectively. In a multivariate analysis, a pretreatment testosterone level in the lower range of normal was the only variable that predicted for delayed testosterone normalization after NAAD (p = 0.00047). Among 45 patients with information concerning androgen deprivation-related symptoms recorded 1 year after cessation of NAAD, 24 (53%) had normalized testosterone levels, but in 21 patients (47%), the levels had not yet returned to normal. At 1 year, only 1 (4%) of 24 patients whose testosterone level had returned to normal experienced NAAD related symptoms compared with 14 (67%) of 21 patients who did not have normal testosterone levels (p <0.001). CONCLUSION: Testosterone levels often remain depressed for extended periods after cessation of short-course NAAD. Lower baseline testosterone levels predict for a delay in testosterone normalization, and the persistence of symptoms related to androgen deprivation correlates with low testosterone levels. PMID- 11872291 TI - Role of hormonal therapy in the management of intermediate- to high-risk prostate cancer treated with permanent radioactive seed implantation. AB - PURPOSE: To study the impact of hormonal therapy (HTx) on intermediate- to high risk prostate cancer treated with permanent radioactive seed implantation. METHODS AND MATERIALS: Patients with Stage T1b-T3bN0 prostate cancer, and Gleason score > or = 7 or prostate-specific antigen (PSA) level >10 ng/mL were treated with seed implantation with or without HTx. Their disease was defined as intermediate risk (PSA 10-20, Gleason score 7, or Stage T2b) or high risk (two or more intermediate criteria, or PSA >20 ng/mL, Gleason score 8-10, or Stage T2c T3). The median follow-up for 201 eligible patients was 42 months (range 18-110). Biochemical failure was defined as a rising PSA >1.0 ng/mL. Pretreatment disease characteristics, implant dose, and HTx were evaluated using univariate and multivariate analyses. RESULTS: HTx significantly improved 5-year actuarial freedom from biochemical failure rate, 79% vs. 54% without HTx. In addition, high dose, PSA < or = 15 ng/mL, intermediate risk, and Stage T2a or lower significantly improved outcome in the univariate analyses. HTx was the most significant predictor of 5-year actuarial freedom from biochemical failure (p <0.0001) in a multivariate analysis. The best outcome was in the intermediate risk patients treated with a high implant dose and HTx, resulting in a 4-year actuarial freedom from biochemical failure rate of 94%. CONCLUSION: In this retrospective review, HTx improved outcome in intermediate- to high-risk prostate cancer patients treated with brachytherapy. HTx was the most important prognostic factor in the univariate and multivariate analyses. PMID- 11872292 TI - Factors influencing risk of acute urinary retention after TRUS-guided permanent prostate seed implantation. AB - PURPOSE: To look for factors predictive of acute urinary retention (AUR) after permanent seed prostate brachytherapy. METHODS AND MATERIALS: From March 1999 to February 2001, 150 permanent seed prostate implants were performed at Princess Margaret Hospital (Stage T1c, n = 113; T2a, n = 37; mean prostate-specific antigen level 5.9 ng/mL, prescription dose 145 Gy per Task Group No. 43). alpha Blockers were used routinely after implantation. Dosimetry was based on the 1 month postimplant CT scan. The International Prostate Symptom Score (IPSS) and catheterization were recorded at 1 month and 3 months and then every 3 months. The following variables were examined: age, baseline IPSS, prior androgen ablation, prostate transrectal ultrasound volume, number of seeds, D(90), V(100), V(200), and urethral dose. RESULTS: Twenty patients (13%) experienced AUR. No difference was seen in the mean D(90) (149 Gy vs. 152 Gy, p = 0.6), V(100) (90% vs. 91%, p = 0.6), V(200) (23% vs. 25% p = 0.4), IPSS (6.4 vs. 5.9, p = 0.8), or maximal urethral dose (204 Gy vs. 210 Gy, p = 0.5). The prostate volume was significantly larger in men with AUR (39.8 cm(3) vs. 34.3 cm(3), p = 0.003), and the mean number of seeds was higher (112 vs. 103, p = 0.006). Of the 20 patients experiencing AUR, 11 (55%) had received prior antiandrogen therapy to downsize their prostates vs. 35 (27%) of the 130 who did not have AUR (p = 0.02). Multivariate analysis showed prostate volume and prior hormone use to be independent predictors of AUR. CONCLUSIONS: Implant quality as determined by D(90), V(100), V(200), and urethral dose did not predict AUR. Prostate size was the major determinant of AUR. For any given prostate size, prior androgen ablation increased the risk of AUR. Men with larger prostates should be aware of the increased risk when contemplating brachytherapy. PMID- 11872294 TI - Use of intraoperative electron beam radiotherapy in the management of retroperitoneal soft tissue sarcomas. AB - PURPOSE: To evaluate the disease control, survival results, and tolerance of intraoperative electron beam radiotherapy (IOERT) as a component of treatment for retroperitoneal soft tissue sarcomas. METHODS AND MATERIALS: Between March 1981 and September 1995, 87 patients with primary (n = 43) or recurrent (n = 44) retroperitoneal or intrapelvic sarcomas received IOERT as a component of treatment at the Mayo Clinic. The tumors were high grade in 54 patients (62%) and low grade in 33 (38%). The median tumor size was 10 cm (range 2-36). All patients underwent maximal surgical resection with IOERT; in 72 patients, only microscopic or no residual tumor remained. The IOERT doses ranged from 8.75 to 30 Gy (median 15). All primary tumors received external beam irradiation (EBRT) with a median dose of 48.6 Gy. Thirty-four of the 44 recurrent tumors received EBRT to a median dose of 45 Gy. All patients were followed prospectively for outcome and toxicity evaluation. RESULTS: The median follow-up, based on 46 patients (53%) currently alive, was 3.5 years. The overall estimated 5-year survival was 47%. For patients with tumors > or = 10 cm, the 5-year overall survival was significantly poorer (28%) than for those with smaller lesions (60%) (p = 0.01). Neither primary vs. recurrent status nor tumor grade had a significant impact on survival. Patients with gross residual tumor had a marginally significantly poorer survival compared with patients with microscopic or no residual tumor, with a 5-year survival rate of 37% and 52%, respectively (p = 0.08). A total of 49 patients (56%) experienced failure, including 20 local recurrences (23%). The median time to failure was 2.3 years. Four recurrences were within the IOERT field, 3 within the IOERT and EBRT field, and 13 within the EBRT field alone. The 3- and 5-year estimated local control rate was 77% and 59%, respectively. Local control was marginally significantly affected by the amount of residual tumor, with a 5-year local control rate of 41% for those with gross residual tumor, 60% for those with microscopic residual tumor, and 100% for those with no residual tumor (p = 0.09). Gastrointestinal complications were recorded in 12 incidences (Grade 3 or higher toxicity). These complications were believed to be secondary to surgery and/or EBRT in 10 of the 12 cases. Seven patients had fistula formation, and 3 experienced severe proctitis. Grade 3 peripheral neurologic toxicities occurred in 9 patients (10%), but none had pain as a component of their neuropathy. CONCLUSION: Retroperitoneal soft tissue sarcomas can be treated with an aggressive combined approach of EBRT, surgery, and IOERT, with acceptable toxicity. Local control in primary disease appears to be improved in this retrospective series with this approach. Distant disease control and options for recurrent disease needs further definition. PMID- 11872293 TI - The dosimetry of prostate brachytherapy-induced urethral strictures. AB - PURPOSE: There is a paucity of data regarding the incidence of urethral strictures after prostate brachytherapy. In this study, we evaluate multiple clinical, treatment, and dosimetric parameters to identify factors associated with the development of brachytherapy-induced urethral strictures. METHODS AND MATERIALS: 425 patients underwent transperineal ultrasound-guided prostate brachytherapy using either (103)Pd or (125)I for clinical T1b/T3a NxM0 (1997, American Joint Committee on Cancer) adenocarcinoma of the prostate gland from April 1995 to October 1999. No patient was lost to follow-up. 221 patients were implanted with (103)Pd and 204 patients with (125)I. The median patient age was 68 years (range 48-81 years). The median follow-up was 35.2 months (range 15-72 months). Follow-up was calculated from the day of implantation. Thirteen patients developed brachytherapy-induced strictures, and all strictures involved the membranous urethra. A control group of 35 patients was rigorously matched to the stricture patients in terms of treatment approach; i.e., choice of isotope, plus or minus radiation therapy, and plus or minus hormonal manipulation. Nine of the 13 stricture patients had detailed Day 0 urethral dosimetry available for review. The apex of the prostate gland and the membranous urethra were defined by CT evaluation. Urethral dosimetry was reported for the prostatic urethra, the apical slice of the prostate gland, and the membranous urethra which was defined as extending 20 mm in length. RESULTS: The 5-year actuarial risk of a urethral stricture was 5.3%, with a median time to development of 26.6 months (range 7.8 44.1 months). Of multiple clinical and treatment parameters evaluated, only the duration of hormonal manipulation (>4 months, p = 0.011) was predictive for the development of a urethral stricture. The radiation dose to the membranous urethra was significantly greater in patients with strictures than those without: 97.6% +/- 20.8% vs. 81.0% +/- 19.8% of prescribed minimum prostate dose, mPD (p = 0.031). The urethral dose 20 mm distal to the prostate apex was 57.6% +/- 23.8% vs. 31.5% +/- 13.9% of mPD for the stricture and control patients, respectively (p = 0.011). In addition, the extent of the 75% mPD and 50% mPD levels beyond the prostatic apex was also significantly greater for stricture patients, 16.6 +/- 5.3 mm vs. 11.9 +/- 4.5 mm (p = 0.010) and 19.0 +/- 3.2 mm vs. 16.0 +/- 3.4 mm (p = 0.021), respectively. Dose to the prostatic urethra was not predictive of stricture, but the magnitude and extent of high-dose regions within the prostate were predictive of stricture. Twelve of the 13 patients who developed urethral strictures were successfully managed by dilatation/transurethral incision. To date, 1 of the 12 patients has required a second dilatation. The remaining patient developed an iatrogenic induced injury and was catheter-dependent for 6 months. CONCLUSIONS: After prostate brachytherapy, the actuarial 5-year incidence of urethral strictures is 5.3% with a median time to development of 26.6 months. All strictures involved the membranous urethra and occurred within the first 44 months after brachytherapy. In most cases, membranous urethral strictures are easily managed with dilatation/incision. Factors predicting for the development of a urethral stricture included the magnitude and extent of high-dose regions within the prostate, the mean membranous urethra dose and the dose 20 mm distal to the prostatic apex, the maximum extent along the membranous urethra of certain dose levels, and the duration of hormonal manipulation. PMID- 11872295 TI - Relationship between acute and late normal tissue injury after postoperative radiotherapy in endometrial cancer. AB - PURPOSE: To evaluate the relationship between acute and late normal tissue reactions in 317 consecutive endometrial cancer patients treated with surgery and adjuvant radiotherapy (RT). METHODS: The data of 317 patients (staging according to the International Federation of Gynecology and Obstetrics) treated with postoperative RT were analyzed. Both low-dose-rate brachytherapy and external beam RT were applied in 247 patients (78%); brachytherapy only in 49 (15%) and external beam irradiation only in 21 (7%). The median follow-up was 7.3 years (range 4-21). The European Organization for Research and Treatment of Cancer, Radiation Therapy Oncology Group system with elements of the late effects of normal tissue, subjective, objective, management, analytic (LENT/SOMA) scale was used to score the RT reactions. The correlation between the occurrence and severity of acute and late bowel and bladder toxicity, as well as the relationship between the severity of acute effects and time to occurrence of late reactions, were assessed using linear and logistic regression analyses. RESULTS: Of the 317 patients, 268 (85%) experienced acute RT reactions of any grade. Severe acute bowel reactions were observed in 15 patients (5%), urinary bladder complications in 1 patient (0.5%), cutaneous in 1 patient (0.5%), and vaginal in 1 patient (0.5%). Severe acute hematologic toxicity was seen in 3 patients (1%). A total of 158 patients (51%) experienced late RT reactions of any grade. Severe late bowel reactions were observed in 19 patients (6%), urinary bladder in 5 (2%), vaginal in 3 (1%), and bone in 10 (4%). When all toxic events were considered, there was a highly significant correlation between the acute and late bowel reactions (p <0.001), but the acute and late urinary bladder reactions did not correlate (p = 0.64). The grade of acute toxicity was found to predict the grade of late toxicity for the bowel but not for the bladder (p <0.001 and p = 0.47, respectively). The severity of acute bowel and bladder toxicity did not correlate with the time to occurrence of late toxicity in these locations (p = 0.34 and p = 0.47, respectively). CONCLUSION: Patients with increased acute bowel toxicity during postoperative RT for endometrial cancer have an increased risk of late bowel injury. A higher grade of acute bowel complications correlated with more severe late events, but was not predictive for its latency time. These findings suggest the possibility of an early indication of patients with an increased risk of late toxicity in whom preventive measures might be attempted. PMID- 11872296 TI - Lethal pulmonary complications significantly correlate with individually assessed mean lung dose in patients with hematologic malignancies treated with total body irradiation. AB - PURPOSE: To assess the impact of lung dose on lethal pulmonary complications (LPCs) in a single-center group of patients with hematologic malignancies treated with total body irradiation (TBI) in the conditioning regimen for bone marrow transplantation (BMT). METHODS: The mean lung dose of 101 TBI-conditioned patients was assessed by a thorough (1 SD around 2%) in vivo transit dosimetry technique. Fractionated TBI (10 Gy, 3.33 Gy/fraction, 1 fraction/d, 0.055 Gy/min) was delivered using a lateral-opposed beam technique with shielding of the lung by the arms. The median lung dose was 9.4 Gy (1 SD 0.8 Gy, range 7.8--11.4). The LPCs included idiopathic interstitial pneumonia (IIP) and non-idiopathic IP (non IIP). RESULTS: Nine LPCs were observed. LPCs were observed in 2 (3.8%) of 52 patients in the group with a lung dose < or = 9.4 Gy and in 7 (14.3%) of 49 patients in the >9.4 Gy group. The 6-month LPC risk was 3.8% and 19.2% (p = 0.05), respectively. A multivariate analysis adjusted by the following variables: type of malignancy (acute leukemia, chronic leukemia, lymphoma, myeloma), type of BMT (allogeneic, autologous), cytomegalovirus infection, graft vs. host disease, and previously administered drugs (bleomycin, cytarabine, cyclophosphamide, nitrosoureas), revealed a significant and independent association between lung dose and LPC risk (p = 0.02; relative risk = 6.7). Of the variables analyzed, BMT type (p = 0.04; relative risk = 6.6) had a risk predictive role. CONCLUSION: The mean lung dose is an independent predictor of LPC risk in patients treated with the 3 x 3.33-Gy low-dose-rate TBI technique. Allogeneic BMT is associated with a higher risk of LPCs. PMID- 11872297 TI - Radiotherapy in age-related macula degeneration. AB - PURPOSE: To ascertain the benefit from radiotherapy in age-related macula degeneration in a single-arm longitudinal study. METHODS AND MATERIALS: From 1997 to 1998, 39 patients with occult and 33 patients with classic choroidal neovascularization (CNV) were irradiated with 16 Gy. Fluorescein angiography and measurements of visual acuity were performed before and 3, 6, and 12 months after irradiation. RESULTS: Complete follow-up data for 1 year were available from 69 patients. The mean patient age was 72 years (range 49-92). Vision decreased in 43, was stable in 18, and improved in 8 cases. The mean vision deteriorated significantly (p = 0.02, Wilcoxon test), particularly within the first 3 months. Patients with occult CNV did significantly better than did those with classic CNV (p = 0.03). The proportion of patients retaining vision > or = 0.2 fell from 65% to 42% (p <0.01), for classic and occult CNV from 50% to 23%, and for occult CNV from 77% to 56% (p < 0.02), respectively. CNV size increased in 30 patients and was stable in 38. Neither age (p = 0.17) nor gender (p = 0.21, chi-square test) influenced prognosis. Four patients reported transitional complaints. CONCLUSION: Low-dose fractionated radiotherapy with 16 Gy is well tolerated. However, vision and reading ability were not preserved in most patients. PMID- 11872298 TI - Consensus guidelines for radiation therapy of benign diseases: a multicenter approach in Germany. AB - PURPOSE: To overcome the lack of written guidelines for radiation therapy (RT) of benign diseases, the German Working Group on Radiotherapy of Benign Diseases initiated a consensus process in 1999 to warrant continuous quality assurance and outcome research in this field. METHODS: An expert panel was convened to define key issues and develop written guidelines for RT of benign diseases. Pertinent information and data from published literature were reviewed, and data of most importance were identified. In addition, a patterns of care study was conducted to obtain a nationwide survey on the current status and treatment standards. RESULTS: From the data gathered, the expert panel prepared a first consensus statement that was open to propositions and comments from all participating institutions. After completion of the multicenter discussion, a final written consensus statement was compiled, discussed, and finally agreed on during a national conference of radiation therapists. For each individual nonmalignant disease, the accepted RT concepts were documented. Finally, specific evaluation tools and recommendations for follow-up examinations were defined. CONCLUSIONS: For the first time, written consensus guidelines for RT of nonmalignant diseases have been developed by the interaction of all institutions involved. These guidelines may serve as a starting point for quality assessment, prospective clinical trials, and outcome research. PMID- 11872299 TI - Relationship between cyclin D1 expression and poor radioresponse of murine carcinomas. AB - PURPOSE: We recently reported that overexpression of epidermal growth factor receptor (EGFR) positively correlated with radioresistance of murine carcinomas. Because cyclin D1 is a downstream sensor of EGFR activation, the present study investigated whether a relationship exists between the extent of cyclin D1 expression and in vivo radiocurability of murine tumors. We further investigated the influence of radiation on cyclin D1 expression and the expression of p27, an inhibitor of the cyclin D1 downstream pathway, as well as the relationship of these molecular determinants to cell proliferation and induced apoptosis in tumors exposed to radiation. METHODS AND MATERIALS: Cyclin D1 expression was assayed in nine carcinomas syngeneic to C3Hf/Kam mice using Western blot analysis. These tumors greatly differed in their radioresponse as assessed by TCD(50). The expression of cyclin D1 and p27 proteins was determined by Western blotting. Cell proliferative activity in tumors was determined by proliferating cell nuclear antigen (PCNA) immunochemistry. The effect of irradiation on the expression of cyclin D1 or p27 proteins and on PCNA positivity was determined in the radiosensitive OCa-I and in the radioresistant SCC-VII tumors. RESULTS: Cyclin D1 expression varied among tumors by 40-fold, and its magnitude positively correlated with poorer tumor radioresponse (higher TCD(50) values). The level of cyclin D1 expression paralleled that of EGFR. A 15-Gy dose reduced constitutive expression of cyclin D1 in the radiosensitive OCa-I tumors, but had no influence on expression of cyclin D1 in the radioresistant SCC-VII tumors. In contrast, 15 Gy increased the expression of p27 in radiosensitive tumors and reduced it in radioresistant tumors. Radiation induced no significant apoptosis or change in the percentage of PCNA-positive (proliferating) cells in SCC-VII tumors with high cyclin D1 levels, but it induced significant apoptosis and a decrease in the percentage of proliferating cells in OCa-I tumors with low cyclin D1 expression. CONCLUSION: Our findings show a positive correlation between cyclin D1 expression and tumor radioresistance. The expression of cyclin D1 and p27 was modified by radiation and was associated with cellular response to radiation, but this depended on the pretreatment level of cyclin D1 expression. These findings may have important clinical implications: The pretreatment assessment of cyclin D1 expression could serve as a useful predictor of radiotherapy outcome and assist in selecting an effective treatment modality. PMID- 11872300 TI - Evaluation of respiratory movement during gated radiotherapy using film and electronic portal imaging. AB - PURPOSE: To evaluate the effectiveness of a commercial system(1) in reducing respiration-induced treatment uncertainty by gating the radiation delivery. METHODS AND MATERIALS: The gating system considered here measures respiration from the position of a reflective marker on the patient's chest. Respiration triggered planning CT scans were obtained for 8 patients (4 lung, 4 liver) at the intended phase of respiration (6 at end expiration and 2 at end inspiration). In addition, fluoroscopic movies were recorded simultaneously with the respiratory waveform. During the treatment sessions, gated localization films were used to measure the position of the diaphragm relative to the vertebral bodies, which was compared to the reference digitally reconstructed radiograph derived from the respiration-triggered planning CT. Variability was quantified by the standard deviation about the mean position. We also assessed the interfraction variability of soft tissue structures during gated treatment in 2 patients using an amorphous silicon electronic portal imaging device. RESULTS: The gated localization films revealed an interfraction patient-averaged diaphragm variability of 2.8 +/- 1.0 mm (error bars indicate standard deviation in the patient population). The fluoroscopic data yielded a patient-averaged intrafraction diaphragm variability of 2.6 +/- 1.7 mm. With no gating, this intrafraction excursion became 6.9 +/- 2.1 mm. In gated localization films, the patient-averaged mean displacement of the diaphragm from the planning position was 0.0 +/- 3.9 mm. However, in 4 of the 8 patients, the mean (over localization films) displacement was >4 mm, indicating a systematic displacement in treatment position from the planned one. The position of soft tissue features observed in portal images during gated treatments over several fractions showed a mean variability between 2.6 and 5.7 mm. The intrafraction variability, however, was between 0.6 and 1.4 mm, indicating that most of the variability was due to patient setup errors rather than to respiratory motion. CONCLUSIONS: The gating system evaluated here reduces the intra- and interfraction variability of anatomy due to respiratory motion. However, systematic displacements were observed in some cases between the location of an anatomic feature at simulation and its location during treatment. Frequent monitoring is advisable with film or portal imaging. PMID- 11872301 TI - Imaging of 1.0-mm-diameter radiopaque markers with megavoltage X-rays: an improved online imaging system. AB - PURPOSE: To improve an online portal imaging system such that implanted cylindrical gold markers of small diameter (no more than 1.0 mm) can be visualized. These small markers would make the implantation procedure much less traumatic for the patient than the large markers (1.6 mm in diameter), which are usually used today to monitor prostate interfraction motion during radiation therapy. METHODS AND MATERIALS: Several changes have been made to a mirror-video based online imaging system to improve image quality. First, the conventional camera tube was replaced by an avalanche-multiplication-based video tube. This new camera tube has very high gain at the target such that the camera noise, which is one of the main causes of image degradation of online portal imaging systems, was overcome and effectively eliminated. Second, the conventional linear accelerator (linac) target was replaced with a low atomic number (low-Z) target such that more diagnostic X-rays are present in the megavoltage X-ray beam. Third, the copper plate buildup layer for the phosphor screen was replaced by a thin plastic layer for detection of the diagnostic X-ray components in the beam generated by the low-Z target. RESULTS: Radiopaque fiducial gold markers of different sizes, i.e., 1.0 mm (diameter) x 5 mm (length) and 0.8 mm (diameter) x 3 mm (length), embedded in an Alderson Rando phantom, can be clearly seen on the images acquired with our improved system. These markers could not be seen on images obtained with any commercial system available in our clinic. CONCLUSION: This work demonstrates the visibility of small-diameter radiopaque markers with an improved online portal imaging system. These markers can be easily implanted into the prostate and used to monitor the interfraction motion of the prostate. PMID- 11872302 TI - A comparison between tandem and ovoids and interstitial gynecologic template brachytherapy dosimetry using a hypothetical computer model. AB - PURPOSE: To evaluate the dose distribution within the clinical target volume between two gynecologic brachytherapy systems---the tandem and ovoids and the Syed-Neblett gynecologic template---using a hypothetical computer model. METHODS AND MATERIALS: Source positions of an intracavitary system (tandem and ovoids) and an interstitial system (GYN template) were digitized into the Nucletron Brachytherapy Planning System. The GYN template is composed of a 13-catheter implant (12 catheters plus a tandem) based on the Syed-Neblett gynecologic template. For the tandem and ovoids, the dwell times of all sources were evenly weighted to produce a pear-shaped isodose distribution. For the GYN template, the dwell times were determined using volume optimization. The prescribed dose was then normalized to point A in the intracavitary system and to a selected isodose line in the interstitial system. The treated volume in the two systems was kept approximately the same, and a cumulative dose-volume histogram of the treated volume was then generated with the Nucletron Brachytherapy Planning System to use for comparison. To evaluate the dose to a hypothetical target, in this case the cervix, a 2-cm-long, 3-cm-diameter cylinder centered along the tandem was digitized as the clinical target volume. The location of this hypothetical cervix was based on the optimal application of the brachytherapy system. A visual comparison of clinical target coverage by the treated volume on three different orthogonal planes through the treated volume was performed. The percentage dose volume histograms of the target were generated for comparison. Multiple midline points were also placed at 5-mm intervals away from the tandem in the plane of the cervix to simulate the location of potential bladder and rectal dose points. Doses to these normal structures were calculated for comparison. RESULTS: Although both systems covered the hypothetical cervix adequately, the interstitial system had a better coverage of the region lateral to the cervix. Smaller volumes of the vagina and uterine fundus received the full dose from the interstitial implant. The cumulative dose-volume histograms revealed larger high dose regions within the treatment volume for the intracavitary system. The volumes receiving > or = 180% of the prescription dose were 31 cc and 17 cc for the intracavitary system and interstitial system, respectively. The isodose lines showed that most of this difference results from the high-dose region around the tandem. The percentage dose-volume histograms showed that a larger percentage of cervix received a higher dose in the intracavitary system. Fifty-two percent of the target volume received 200% or higher of the prescription dose with tandem and ovoids, compared with only 20% with the template system. Analysis of dose points outside of the 100% isodose lines showed a slightly more rapid dose drop off with the interstitial system compared to the intracavitary system. Point doses at 20, 25, and 30 mm from the tandem in the interstitial system were 100%, 69%, and 51% of prescribed dose, and from the intracavitary system were 101%, 76%, and 58%, respectively. CONCLUSIONS: Our dosimetric analysis revealed a better coverage in the parametrial regions, but underdosage of the central cervical region, for the interstitial system. On the other hand, because of the increased distance of source to dose point, there is a more rapid dose drop-off outside the treated volume with the interstitial system, which has the potential to improve tissue sparing. Based on this analysis, we caution against using a radiotherapy system with a homogeneous central dose distribution when treating cervical cancer with an intact uterus. We recommend differential loading of the implant catheters with the majority of dose delivered from the tandem when using an interstitial GYN template with remote afterloader. PMID- 11872303 TI - Prostate brachytherapy postimplant dosimetry: a comparison of prostate quadrants. AB - PURPOSE: To investigate postimplant dosimetry for different regions of the prostate gland in patients treated with transperineal 125Iodine brachytherapy implants for low- and intermediate-risk prostate cancer. METHODS AND MATERIALS: Two hundred eighty-four patients treated with permanent interstitial prostate brachytherapy comprised the study population. A nonuniform, urethral-sparing algorithm was used to plan all patients. Prostate contours were outlined on postimplant CT images. Prostate volumes were then divided into four quadrants: anterior-superior quadrant (ASQ), posterior-superior quadrant (PSQ), anterior inferior quadrant (AIQ), and posterior-inferior quadrant (PIQ). Dose-volume histograms (DVHs) were calculated for the whole prostate and each quadrant. RESULTS: The mean postimplant V(100) +/- 95% confidence (the percent prostate volume encompassed within the isodose surface comprising the prescription dose = 144 Gy) for the ASQ was 78.5 +/- 1.9, which was significantly lower than that of the PSQ, AIQ, and PIQ in which the V(100) plus minus 95% confidence values were 94.9 +/- 0.8, 92.6 +/- 1.2, and 98.7 +/- 0.3, respectively. The mean V(100) +/- 95% confidence for the whole prostate was 90.4 +/- 0.8. Mean values for V(150) and D(90) (the minimum dose in Gy received by 90% of the target volume) for the four quadrants and the whole prostate showed similar results. CONCLUSIONS: Underdosed areas of the planning target volume (PTV), if present, were largely confined to the ASQ, which received a significantly lower dose, on average, compared to the other three quadrants of the prostate. PMID- 11872304 TI - Potential of tomotherapy for total scalp treatment. AB - PURPOSE: Total scalp radiotherapy is required in a variety of clinical situations. We compared conventional lateral photon-electron (LPE) technique with tomotherapy (intensity-modulated radiotherapy [IMRT]). METHODS AND MATERIALS: A patient with Merkel cell carcinoma was treated at our institution using conventional treatment techniques. Treatment plans were conducted using conventional three-dimensional treatment planning and IMRT. The clinical target volume included the entire scalp tissue volumes to the surface of underlying cranial bone, as well as superficial and deep neck nodes in the bilateral neck. To provide a consistent comparison between the IMRT and three-dimensional conventional treatment plans, the dose distributions were normalized such that 90% of the target volumes received the prescription dose. RESULTS: Examination of our results revealed an acceptable dose-volume histogram and adequate coverage of the clinical target volume using the conventional LPE technique. The IMRT plan provided a more homogeneous dose to the target volume; however, critical structure doses were uniformly higher than for the conventional treatment plan. CONCLUSIONS: The IMRT plan resulted in a substantial dose to the lens, brain, and orbit, making it clinically unacceptable compared with the LPE technique. Overall, the LPE technique was superior. PMID- 11872305 TI - Definition of a moving gross target volume for stereotactic radiation therapy of stented coronary arteries. AB - PURPOSE: To measure the effect of cardiac motion on coronary artery stent position during the cardiac cycle as a first step toward exploring the feasibility of stereotactic external beam radiation therapy targeted at restenotic stented coronary arteries. METHODS AND MATERIALS: The three dimensional (3D) position of eight coronary artery stents in 8 patients immobilized in a stereotactic body frame was studied noninvasively by single breathhold ECG-gated multislice spiral computed tomography (MSCT) during 10 retrospectively selected phases, equally distributed throughout the R-R interval of consecutive cardiac cycles. The volume encompassing all measured 3D positions of the stent was measured. RESULTS: Stent volumes measured by MSCT closely agreed with measurements by quantitative coronary angiography (r > 0.99). The mean of the maximum 3D stent center of mass displacement between any two phases during the cardiac cycle for all eight coronary arteries was 7.5 mm (range 3.3-20.5 mm) in the lateral direction, 8.6 mm (range 2.7-21.6 mm) in the ventrodorsal direction, and 8.2 mm (range 2.5-19.7 mm) in the craniocaudal direction. As was anticipated, the volume encompassing all measured 3D positions of the stent represented only a fraction of the whole heart volume in all patients, i.e., less than 0.6%. CONCLUSIONS: ECG-gated MSCT allowed the measurement of the volume encompassing multiphase 3D positions of coronary artery stents during the cardiac cycle. This volume, a measure of the cardiac motion effect on coronary artery stent position during the cardiac cycle, represents a moving gross target for stereotactic external beam radiation therapy. PMID- 11872306 TI - Dosimetric intercomparison for two Australasian clinical trials using an anthropomorphic phantom. AB - INTRODUCTION: Many different factors can affect the accurate delivery of dose to the clinical target volume in radiotherapy. This is particularly important in the context of multicenter clinical trials where different equipment and techniques may be used for supposedly identical treatments. A dosimetric intercomparison employing an anthropomorphic phantom (level III dosimetric intercomparison) can be used to check many of the factors that could affect treatment by mimicking the radiotherapy pathway of a patient as closely as possible. METHODS AND MATERIALS: An anthropomorphic phantom (ART) was taken to 18 radiotherapy centers in Australia and New Zealand and treated for two different treatment scenarios based on current clinical trials of the Trans-Tasman Radiation Oncology Group (TROG): a two-field treatment of a carcinoma of the tonsil (TROG 91.01), and a four-field prostate treatment (TROG 96.01). The dose distribution was assessed in two consecutive treatments using thermoluminescence dosimeters (TLDs) placed throughout the target volume and in "critical" structures such as the lens of the eye or the rectum. The study also included a check of absolute dose calibration in a slab phantom (level I dosimetric intercomparison). The influence of a variety of treatment parameters on the dose homogeneity in the target and the measured dose in the target and the critical organs was evaluated. RESULTS: The dose measurements confirmed that in all participating centers the correct dose was delivered to the ICRU reference point (tonsil: 99.8 +/- 2.3%; prostate: 100.9 +/- 1.9% [1 SD]). Also the absolute dose calibration and the mean dose in the target volume were within the specified action levels of plus minus 5% for all participating centers. No influence of shielding, beam modifiers, beam weighting, treatment planning approach (CT, 2D, 3D), and type of equipment used on the dose in the target and its homogeneity could be demonstrated. However, treatment technique and energy used influenced the dose to the critical organs. It was shown that the interpretation of results could be improved by including two complementary treatment scenarios and a level I intercomparison with the level III dosimetric intercomparison. CONCLUSION: The study demonstrated the feasibility of a level III dosimetric intercomparison service at a cost of approximately $1000(US) per center in Australasia. It confirmed that the dose delivered by all participating centers was as intended in the two treatment scenarios chosen. While this provides reassurance to the oncology community and the general public, the service must be extended to all centers and other potentially more complex treatment scenarios. The present study has built the foundation for this by establishing a baseline and action levels and suggesting improvements in phantom design which will be included in future TROG quality assurance exercises. PMID- 11872307 TI - In regard to Kapp et al.: experience with split-course external beam irradiation +/- chemotherapy and integrated (192)Ir high-dose-rate brachytherapy in the treatment of primary carcinomas of the anal canal. IJROBP 2001;49:997--1005. PMID- 11872309 TI - Triple modality (neoadjuvant chemoradiotherapy followed by surgical resection): is it better than definitive chemoradiation therapy in cancer of the thoracic esophagus? PMID- 11872311 TI - Spontaneous motor activity in human infants with iron-deficiency anemia. AB - This study compared spontaneous motor activity in 6-month-old Chilean infants with or without iron-deficiency anemia (IDA) who were otherwise healthy. Activity was assessed in conjunction with polysomnographic recording during an afternoon nap in 11 infants with IDA and 15 with normal hemoglobin levels. All infants were given oral iron, and activity was reassessed at 12 and 18 months. Using actigraphs placed on the ankle, the frequency of movement units per minute was determined for each waking/sleep state. The total amount of time infants were in an alert-active state before and after the nap was used to calculate the proportion of movements/minute of waking. There were no differences between anemic and nonanemic infants in total recording time, duration of sleep, or motor activity during sleep. However, infants with IDA showed reduced motor activity during waking at all ages. The magnitude of the differences increased at 12 and 18 months. Thus, IDA was associated with reduced motor activity in infants even after iron treatment. It will be important to confirm these results in a larger sample and to determine the 24-h pattern of motor activity, since reduced motor activity may limit infants' opportunities to explore and learn from the social and physical environment. PMID- 11872312 TI - The ability of neonatal and maternal erythrocytes to produce reactive oxygen species in response to oxidative stress. AB - The ability of neonatal and maternal erythrocytes to produce reactive oxygen species (ROS) in response to oxidative stress was investigated using the chemiluminescence probe Cypridina luciferin analogue. The chemiluminescence probe, based on 2-methyl-6-[p-methoxyphenyl]-3,7-dihydro-imidazo[1,2 alpha]pyrazin-3-one (MCLA), is highly specific and sensitive to superoxide anion (O(2)(-)) and singlet oxygen (1O(2)). Blood from 11 mothers who experienced no complications and their healthy full-term newborns was collected and heparinized. MCLA was put into the washed erythrocytes suspension, and phenylhydrazine (PH) was added to cause oxidative stress. Chemiluminescence was measured using an Argus 50 image processing system. It was found that erythrocytes in neonatal blood had a 2.0-fold greater maximum chemiluminescence than did those in maternal blood. There was no change in the emission after the addition of NaN(3), but there was complete suppression with superoxide dismutase (SOD), demonstrating that these were O(2)(-). The present results demonstrated that neonatal erythrocytes produce about twice as much O(2)(-) as do adult erythrocytes in response to oxidative stress. With this method, it is very simple to measure the amount of O(2)(-) produced in erythrocytes and the rate at which they are produced. This method may therefore be very useful for determining the effects of antioxidants. PMID- 11872313 TI - Arousal from sleep in infants is impaired following an infection. AB - Numerous studies have postulated a link between recent infection and Sudden Infant Death Syndrome (SIDS). In this study we contrasted arousal responses from sleep in infants on the day of discharge from hospital following an infection with those when fully recovered and also with well age-matched control infants. Thirteen term infants comprised the infection group and nine well infants acted as age-matched controls. All infants were studied using daytime polysomnography and multiple measurements of arousal threshold (cm H(2)O) in response to air-jet stimulation applied alternately to the nares were made in both active sleep (AS) and quiet sleep (QS). All infants were studied on two occasions: firstly, immediately before discharge from the Paediatric ward, and secondly, 10-15 days later when they were completely well in the case of the infection group.Arousal thresholds in QS in the infection group were significantly elevated on the day of discharge (262 +/- 48 cm H(2)O) compared with 10-15 days later (205 +/- 31 cm H(2)O, p<0.05). Thresholds in the control group were not different between studies. This study provides evidence that arousability from QS is impaired following an infection and we postulate that this may explain the increased risk for SIDS following infection observed in previous studies. PMID- 11872314 TI - Amniotic fluid soluble human leukocyte antigen G is markedly decreased in offspring with neural tube defects. AB - Pregnancies affected by a neural tube defect show changes in thymus morphology, neonatal and maternal T-cell repertoire. Amniotic fluid levels of soluble human leukocyte antigen G (HLA-G) (an immuno-modulatory protein) were found to be significantly lower as compared to controls. This may reflect a diminished cell mediated immunity in neural tube defects. PMID- 11872316 TI - Interaction between bed sharing and other sleep environments during the first six months of life. AB - This investigation was carried out to determine the relationship between bed sharing and other places of infant sleep in the first six months of life, and to identify patterns of change in the place of infant sleep for infants who do and do not routinely bed share in the first six months of life. The sleep--wake behaviour and place of infant sleep were recorded, at weekly intervals, for bed sharing (n=25) and non-bed sharing (n=68) infants between 2 and 24 weeks after birth. Bed sharing infants spent a significantly increased proportion of their total sleep time per 24 h in other sleep environments which favoured close parental proximity and significantly less time in solitary sleep. Non-bed sharing infants spent a substantial proportion of their time sleeping alone from 2 weeks of age whereas the transition to sleeping alone occurred after 16 weeks for bed sharing infants. We have found that bed sharing acts as a proxy for increased close parental proximity during the first six months of life. This may be of significance in studies which examine the relationship between bed sharing and sudden infant death syndrome. PMID- 11872315 TI - A review of the anatomy of the upper airway in early infancy and its possible relevance to SIDS. AB - BACKGROUND: Since the danger of prone sleeping in the first 6 months of life has been publicised, there has been a dramatic and consistent reduction in the incidence of sudden infant death syndrome (SIDS). However, unexpected infant deaths and apparent life-threatening events (ALTEs) continue to occur that are clearly not associated with known epidemiological risk factors. AIMS: To review the unique features of the anatomy and function of the upper airway of the young infant which contribute to increased vulnerability to hypoxia in this age group. We discuss the clinical identification of those infants at risk of obstruction or restriction of the upper airway and the management of the 'at risk' infant. CONCLUSIONS: In the era after the "back to sleep" campaigns, it is likely that an increasing proportion of cases of ALTEs and SIDS will be related to obstruction or limitation of upper airway size leading to sleep hypoxia/asphyxia. This type of problem may be anticipated by evaluation and investigation of infants with signs or a clinical history consistent with possible upper respiratory tract compromise, including micrognathia. PMID- 11872317 TI - Rescue high frequency oscillation and predictors of adverse neurodevelopmental outcome in preterm infants. AB - BACKGROUND: High frequency oscillation (HFO) is now frequently used as rescue support, but it has been suggested that as many as one-third of survivors have abnormal neurodevelopmental findings at follow-up. OBJECTIVE: To identify risk factors for adverse neurodevelopmental outcome at 1 and 2 years in very prematurely born patients, who, because of severe neonatal respiratory failure, had required transfer to high frequency oscillation (HFO). METHODS: A case control study was performed. Controls were supported by conventional mechanical ventilation (CMV) only and matched to HFO infants for gestational age. At 1 and 2 years, neurodevelopmental status was assessed in both groups. Abnormal neurodevelopmental outcome was diagnosed if infants had impairment with or without disability or a Griffiths developmental quotient of at least two standard deviations below the mean. PATIENTS: Fifty-six infants were studied, median gestation age of 28 weeks (range 23--31). RESULTS: At 2 years of age, a greater proportion of the HFO infants compared to the controls had an abnormal outcome (p<0.05). HFO infants with an abnormal outcome compared to those with a normal outcome had poorer oxygenation prior to transfer to HFO (p=0.05), but did not have a lower initial improvement in oxygenation or longer duration of hypocarbia on HFO. Logistic regression demonstrated adverse outcomes significantly related to HFO use and gestational age in the whole study population and to gestational age in the HFO infants. CONCLUSION: An initial response to HFO does not guarantee normal neurodevelopmental outcome. Rescue HFO in very immature infants should be used cautiously. PMID- 11872318 TI - Signals of adiposity. AB - Adipose tissue, a reserve of energy, has played an essential role in mammalian evolution. Adipose tissue differs from other tissues in that its mass has considerable capacity to expand, which while beneficial in decreasing the risk of starvation, increases the risk of predation. Adipose tissue mass is thus under tight control in nondomestic species. Adipose tissue secretes a variety of factors, some of which (leptin, tumor necrosis factor (TNF) alpha, resistin) are thought to be involved in modulation of adipose mass. Leptin has a variety of functions, primarily targetting the hypothalamus where it acts to decrease appetite and increase energy expenditure. Leptin is also involved in the adaptations to fasting, and leptin is also required for normal reproductive and immune function. TNF alpha and resistin appear to have key paracrine roles, attenuating the anabolic effects of insulin on adipose tissue metabolism. PMID- 11872319 TI - Leptin and the regulation of food intake, energy homeostasis and immunity with special focus on periparturient ruminants. AB - The biology of leptin has been studied most extensively in rodents and in humans. Leptin is involved in the regulation of food intake, energy homeostasis and immunity. Leptin is primarily produced in white adipose tissue and acts via a family of membrane bound receptors, including an isoform with a long intracellular domain (OB-Rb), and many isoforms with short intracellular domains (Ob-Rs). OB-Rb is predominantly expressed in the hypothalamic regions involved in the regulation of food intake and energy homeostasis. The other isoforms are distributed ubiquitously and are found in most peripheral tissues in far greater abundance than OB-Rb. The effects of leptin on food intake and energy homeostasis are central and are mediated via a network of orexigenic neuropeptides (neuropeptide Y, galanin, galanin-like peptide, melanin-concentrating hormone, orexins, agouti-related peptide) and anorexigenic neuropeptides (corticotropin releasing hormone, pro-opiomelanocortin, alpha-melanocyte stimulating hormone and cocaine- and amphetamine-regulated transcript). In addition, leptin acts directly on immune cells to stimulate hematopoesis, T-cell immunity, phagocytosis, cytokine production, and to attenuate susceptibility to infectious insults. Emerging data in ruminants suggest that leptin is dynamically regulated by many factors and physiological states. Thus, leptin is secreted in a pulsatile fashion, but without a marked diurnal rhythm. A positive relationship between adiposity and plasma leptin concentration exists in growing and lactating ruminants. The concentration of plasma leptin increases during pregnancy, starts to decline 1--2 wk before parturition, and reaches a nadir in early lactation. The reduction of plasma leptin at parturition is likely to promote centrally mediated adaptations required in periods of energy deficit, but could have negative effects on immune cell function. Future research is needed in ruminants to address the roles played by leptin and the central nervous system in orchestrating metabolism during the periparturient period and during infectious diseases. PMID- 11872320 TI - Leptin: a possible metabolic signal affecting reproduction. AB - Since its discovery in 1994, leptin, a protein hormone synthesized and secreted by adipose tissue, has been shown to regulate feed intake in several species including sheep and pigs. Although a nimiety of information exists regarding the physiological role of leptin in rodents and humans, the regulation and action of leptin in domestic animals is less certain. Emerging evidence in several species indicates that leptin may also affect the hypothalamo-pituitary-gonadal axis. Leptin receptor mRNA is present in the anterior pituitary and hypothalamus of several species, including sheep. In rats, effects of leptin on GnRH, LH and FSH secretion have been inconsistent, with leptin exhibiting both stimulatory and inhibitory action in vivo and in vitro. Evidence to support direct action of leptin at the level of the gonad indicates that the leptin receptor and its mRNA are present in ovarian tissue of several species, including cattle. These leptin receptors are functional, since leptin inhibits insulin-induced steroidogenesis of both granulosa and thecal cells of cattle in vitro. Leptin receptor mRNA is also found in the testes of rodents. As with the ovary, these receptors are functional, at least in rats, since leptin inhibits hCG-induced testosterone secretion by Leydig cells in vitro. During pregnancy, placental production of leptin may be a major contributor to the increase in maternal leptin in primates but not rodents. However, in both primates and rodents, leptin receptors exist in placental tissues and may regulate metabolism of the fetal-placental unit. As specific leptin immunoassays are developed for domestic animals, in vivo associations may then be made among leptin, body energy stores, dietary energy intake and reproductive function. This may lead to a more definitive role of leptin in domestic animal reproduction. PMID- 11872321 TI - Leptin in ruminants. Gene expression in adipose tissue and mammary gland, and regulation of plasma concentration. AB - This paper reviews data on leptin gene expression in adipose tissue (AT) and mammary gland of adult ruminants, as well as on plasma leptin variations, according to genetic, physiological, nutritional and environmental factors. AT leptin mRNA level was higher in sheep and goat subcutaneous than visceral tissues, and the opposite was observed in cattle; it was higher in fat than in lean selection line in sheep; it was decreased by undernutrition and increased by refeeding in cattle and sheep, and not changed by adding soybeans to the diet of lactating goats; it was increased by injection of NPY to sheep, and by GH treatment of growing sheep and cattle. Insulin and glucocorticoids in vitro increased AT leptin mRNA in cattle, and leptin production in sheep. Long daylength increased AT lipogenic activities and leptin mRNA, as well as plasma leptin in sheep. Mammary tissue leptin mRNA level was high during early pregnancy and was lower but still expressed during late pregnancy and lactation in sheep. Leptin was present in sheep mammary adipocytes, epithelial and myoepithelial cells during early pregnancy, late pregnancy and lactation, respectively. Plasma leptin in cattle and sheep was first studied thanks to a commercial "multi species" kit. It was positively related to body fatness and energy balance or feeding level, and decreased by beta-agonist injection. The recent development of specific RIA for ruminant leptin enabled more quantitative study of changes in plasma leptin concentration, which were explained for 35--50% by body fatness and for 15--20% by feeding level. The response of plasma leptin to meal intake was related positively to glycemia, and negatively to plasma 3-hydroxybutyrate. The putative physiological roles of changes in leptin gene expression are discussed in relation with published data on leptin receptors in several body tissues, and on in vivo or in vitro effects of leptin treatment. PMID- 11872322 TI - Biology of leptin in the pig. AB - The recently discovered protein, leptin, which is secreted by fat cells in response to changes in body weight or energy, has been implicated in regulation of feed intake, energy expenditure and the neuroendocrine axis in rodents and humans. Leptin was first identified as the gene product found deficient in the obese ob/ob mouse. Administration of leptin to ob/ob mice led to improved reproduction as well as reduced feed intake and weight loss. The porcine leptin receptor has been cloned and is a member of the class 1 cytokine family of receptors. Leptin has been implicated in the regulation of immune function and the anorexia associated with disease. The leptin receptor is localized in the brain and pituitary of the pig. The leptin response to acute inflammation is uncoupled from anorexia and is differentially regulated among swine genotypes. In vitro studies demonstrated that the leptin gene is expressed by porcine preadipocytes and leptin gene expression is highly dependent on dexamethasone induced preadipocyte differentiation. Hormonally driven preadipocyte recruitment and subsequent fat cell size may regulate leptin gene expression in the pig. Expression of CCAAT-enhancer binding proteinalpha (C/EBPalpha) mediates insulin dependent preadipocyte leptin gene expression during lipid accretion. In contrast, insulin independent leptin gene expression may be maintained by C/EBPalpha auto-activation and phosphorylation/dephosphorylation. Adipogenic hormones may increase adipose tissue leptin gene expression in the fetus indirectly by inducing preadipocyte recruitment and subsequent differentiation. Central administration of leptin to pigs suppressed feed intake and stimulated growth hormone (GH) secretion. Serum leptin concentrations increased with age and estradiol-induced leptin mRNA expression in fat was age and weight dependent in prepuberal gilts. This occurred at the time of expected puberty in intact contemporaries and was associated with greater LH secretion. Further work demonstrated that leptin acts directly on pituitary cells to enhance LH and GH secretion, and brain tissue to stimulate gonadotropin releasing hormone secretion. Thus, development of nutritional schemes and (or) gene therapy to manipulate leptin secretion will lead to practical methods of controlling appetite, growth and reproduction in farm animals, thereby increasing efficiency of lean meat production. PMID- 11872323 TI - Chicken leptin: properties and actions. AB - Chicken leptin cDNA shows a high homology to mammalian homologous, with an expression localized in the liver and adipose tissue. It is noteworthy, that the hepatic expression is most likely associated with the primary role that this organ plays in lipogenic activity in avian species. As in mammals, chicken leptin expression is regulated by hormonal and nutritional status. This regulation is tissue-specific and with a high sensitivity in the liver compared to adipose tissue. The blood leptin levels are regulated by the nutritional state with high levels in the fed state compared to the fasted state. The recombinant chicken leptin markedly inhibits food intake as reported in mammals, suggesting the presence of an hypothalamic leptin receptor. The chicken leptin receptor has been identified and all functional motifs are highly conserved compared to mammalian homologous. Chicken leptin receptor is expressed in the hypothalamus but also in other tissues such as pancreas, where leptin inhibits insulin secretion and thus may have a key role in regulating nutrient utilization in this species. PMID- 11872324 TI - Human CYP3A4 and the metabolism of nefazodone and hydroxynefazodone by human liver microsomes and heterologously expressed enzymes. AB - Nefazodone is an antidepressant with a relatively unique structure and mechanism of action. The current study was conducted to assess the potential for nefazodone to have metabolic drug interactions associated with cytochrome P450 (CYP) enzymes. Nefazodone is metabolised to hydroxynefazodone (OH-NEF), triazoledione (TD), and m-chlorophenylpiperazine (m-CPP), and OH-NEF is metabolised to TD and m CPP. Correlations with enzyme activities in a panel of microsomes prepared from human livers, incubations with heterologously expressed human CYP enzymes, and incubations with enzyme inhibitors were used to study these metabolic pathways. The results suggest that the metabolism of NEF and OH-NEF to each of their active metabolites is catalysed mainly by CYP3A4, which is in agreement with clinical reports of drug--drug interactions of nefazodone with substrates and inhibitors of CYP3A4. PMID- 11872325 TI - An integrative pharmacokinetic and pharmacodynamic study of the 5-HT1A receptor antagonist (S)-UH-301 in the rat. AB - (S)-UH-301 ((-)-(S)-5-fluoro-8-hydroxy-2-(di-n-propylamino)tetralin hydrochloride) is a well known 5-HT(1A) receptor antagonist. The present study describes the pharmacokinetic properties of (S)-UH-301 after subcutaneous administration in rats, using a newly developed HPLC-UV bioanalytical method. The relationships between (S)-UH-301 concentrations and some pharmacodynamic effects were also studied. The AUC of (S)-UH-301 in brain, but not in plasma, increased in proportion to dose (1-100 mumol/kg). However, at doses above 32 mumol/kg, peak concentrations of the drug did not increase in proportion to dose, and there was a doubling of its apparent half-life. There was a good correspondence between the time courses for the antagonism of 8-OH-DPAT-induced motor behaviours and hypothermia and the tissue concentrations of (S)-UH-301. Doses of (S)-UH-301 above 10 mumol/kg decreased 5-HT and dopamine synthesis. Therefore, a selective 5 HT(1A) antagonistic dose range of (S)-UH-301 should be 0.1-10 mumol/kg s.c., corresponding to concentrations below approximately 10 nmol/g in brain and approximately 1 nmol/ml in plasma. PMID- 11872326 TI - Modulation of cholestasis-induced antinociception by CCK receptor agonists and antagonists. AB - Acute cholestasis is associated with increased activity of the endogenous opioid system. Agonists and antagonists of cholecystokinin (CCK) receptors are known to modulate opioid-induced antinociception. In the present study, the effect of the CCK receptor agonist caerulein and the antagonist proglumide on antinociception induced during acute cholestasis was investigated in rats using the tail-flick test. A significant increase in nociception threshold was observed in bile duct ligated (BDL) rats compared to sham-operated controls that was maximum on day 7 after the operation and decreased thereafter. Proglumide (40 mg/kg, i.p.) did not affect nociception in unoperated and sham-operated animals, but exerted a significant potentiation of antinociception in cholestatic rats in a way similar to its potentiation effect on unoperated morphine-treated (2 mg/kg, s.c.) animals. Caerulein (0.005, 0.001, 0.01 and 0.02 mg/kg, s.c.), which did not change nociception per se or in sham-operated animals, also significantly potentiated the antinociception in BDL rats as well as in morphine-treated unoperated controls. Caerulein-induced potentiation of antinociception in BDL animals was completely reversed by proglumide pretreatment. Our findings show that, in cholestatic animals, modulation of nociception by the CCK system is different from normal subjects and resembles the state observable in morphine administered subjects. PMID- 11872327 TI - Successful treatment of reboxetine-induced urinary hesitancy with tamsulosin. AB - Urinary hesitancy can be an uncomfortable side effect during antidepressant treatment. Clinicians often use the selective alpha(1A)-adrenoceptor antagonist, tamsulosin, to treat urinary hesitancy associated with prostate enlargement. We report here a series of case studies in which tamsulosin has been successfully used in the management of urinary hesitancy during therapy with the selective noradrenaline reuptake inhibitor reboxetine for major depressive disorder (MDD). Eight male adults (aged 43-64 years; DSM-IV diagnosis of MDD) who were receiving treatment with reboxetine (4-8 mg/day) were considered candidates for concomitant tamsulosin (0.4 mg/day) therapy. Tamsulosin was administered either as prophylaxis (n=4) or as treatment (n=4) for emergent urinary hesitancy. All patients experienced relief of urinary hesitancy within 20 min of tamsulosin therapy and this effect was sustained. Concomitant treatment with tamsulosin should be considered for those patients in whom urinary hesitancy may lead to withdrawal from reboxetine therapy. PMID- 11872328 TI - Effects of acute tryptophan depletion on cognitive function in euthymic bipolar patients. AB - Decreasing brain 5-HT levels by acute tryptophan depletion has been shown to selectively impair cognition in healthy volunteers. In bipolar disorder, ATD causes measurable neurophysiological effects without altering mood. The purpose of this study was to examine the effects of ATD on neuropsychological performance in 14 euthymic bipolar patients. Cognitive function was evaluated 4-6 h after ingestion of a control or depleting amino-acid drink. Plasma tryptophan levels fell significantly following the depleting drink, however there were no main effects on the ID/ED set-shift task, Paired Associates Learning or Vigil. A trend towards a decrease in the proportion of perfect solutions on the Tower of London task was observed when depleted. While ATD reduces 5-HT levels in the brain, it does not appear to alter neuropsychological performance on tests of sustained attention or associative learning. Effects on specific 'executive' functions are less clear, and should be the focus for future research. PMID- 11872329 TI - Role of family history and 5-HTTLPR polymorphism in female seasonal affective disorder patients with and without premenstrual dysphoric disorder. AB - Seasonal affective disorder (SAD) and premenstrual dysphoric disorder (PMDD) share many clinical features, and have been associated with brain serotonin dysfunction. Females with SAD frequently fulfil the diagnostic criteria for PMDD. A polymorphism in the serotonin transporter promoter gene (5-HTTLPR) has been associated with SAD. We investigated the role of family history and 5-HTTLPR in female SAD patients with and without PMDD. Forty-four SAD females with, and 43 SAD females without PMDD, were genotyped for 5-HTTLPR. Family history of affective disorders in first degree relatives was assessed. An association between the presence of PMDD and family history (P=0.0029) and 5-HTTLPR long/short allele-heterozygosity (P=0.033) was found in females with SAD. PMDD and SAD may share genetic vulnerability factors, one candidate gene being 5 HTTLPR. The elevated rate of affective disorders in relatives of patients with SAD and PMDD suggests higher genetic vulnerability in this subgroup when compared to patients with SAD alone. PMID- 11872330 TI - Low platelet monoamine oxidase activity in Swedish imprisoned criminal offenders. AB - Numerous studies report a connection between low platelet monoamine oxidase activity (trbc MAO) and personality traits such as impulsiveness and sensation seeking. Generally, criminal offenders constitute a group of individuals that are high in such temperamental characteristics. In this study, we investigated trbc MAO activity in imprisoned criminal offenders and in controls where the confounding factor of smoking was under control. Radiometric MAO assays were performed in 99 male criminal offenders and in 60 non-criminal volunteers. Offenders had significantly lower trbc MAO activity than controls, i.e., 8.8 +/- 3.0 nmol/10(10) platelets/mm and 11.3 +/- 5.1, respectively (p<0.0001). When only smoking individuals were included in the analysis, the difference in trbc MAO was still statistically significant (p<0.05). Based on these data, we suggest that trbc MAO is related to mechanisms predisposing for development of specific personality characteristics that in turn increase vulnerability for criminal behaviour. The results also suggest that low trbc MAO activity in criminal offenders is not an artefact of cigarette smoking. PMID- 11872331 TI - Schizophrenic patients who smoke have a faster finger tapping rate than non smokers. AB - The increased rate of smoking in schizophrenia patients remains unexplained and may reflect attempts at self-treatment. The effect sought from smoking may be related to nicotine's stimulating action. We tested this hypothesis by examining the relationship between smoking status and finger tapping rate, a measure of central processing, in schizophrenia patients treated with atypical antipsychotics. Smokers showed significantly faster finger tapping rates than non smokers. This was not related to clinical state, illness chronicity, medication side-effects, antipsychotic dose or plasma concentrations. Nicotine can improve central processing in medicated schizophrenia patients and this may constitute part of the incentive for smoking. PMID- 11872332 TI - Functioning and neuronal viability of the anterior cingulate neurons following antipsychotic treatment: MR-spectroscopic imaging in chronic schizophrenia. AB - Magnetic resonance spectroscopic imaging provides a non-invasive approach for testing the hypothesis that neuronal function can improve under atypical antipsychotic medication leading to improvement in cognitive function. We studied two groups of schizophrenic patients, one treated exclusively with typical neuroleptics, the other with atypical medications. 1H MR-spectroscopic imaging of the anterior cingulate gyrus was performed in all patients. Perseveration errors in the Wisconsin Card Sorting Test (WCST) served as an additional marker for cingulate gyrus function. Our results showed that N-acetylaspartate (NAA), a measure of neuronal function, was closely correlated with perseveration errors seen on the WCST. Patients treated with atypical medications had fewer errors on the WCST and higher NAA levels than those on typical medications, and there was a correlation between the time treated with atypical medication, higher NAA levels and better test performance. These results suggest that atypical antipsychotics modify the function of anterior cingulate neurons in a specific manner. PMID- 11872333 TI - Suicide-attempters having immunoglobulin G with affinity for dopamine in cerebrospinal fluid. AB - Altered immunological functions and changes in the monoaminergic neurotransmitter systems are two important observations made previously in the study of possible etiological and pathophysiological factors for psychiatric disorders. In search of tentative autoimmune mechanisms involved in these disorders we studied the presence of immunoglobulin G (DA-IgG) with affinity for the monoamine dopamine (DA) in cerebrospinal fluid (CSF) using ELISA. In CSF from 49 suicide attempters the titer of DA-IgG was significantly higher (P<0.001) than in CSF obtained from control subjects undergoing neurological investigation. The results in the present study indicate that an autoimmune mechanism may be involved in the dopaminergic neurotransmitter system and may be of pathophysiological importance in psychiatric disorders connected to an attempt of suicide. PMID- 11872334 TI - In the rat forced swimming test, NA-system mediated interactions may prevent the 5-HT properties of some subacute antidepressant treatments being expressed. AB - In the rat forced swimming test (FST), reuptake inhibitors selective of either serotonin (5-HT) or noradrenaline (NA) decrease immobility duration, and increase, respectively, swimming and climbing behaviour. In this study, an almost total 6-OHDA-induced NA-depletion prevented the behavioural effects of desipramine, but not fluoxetine. Interestingly, the serotonin/noradrenaline reuptake-inhibitor milnacipran, as well as a (desipramine+fluoxetine) combination, could produce both swimming and climbing behaviour in NA-lesioned rats, but not in non-lesioned. The new antidepressant mirtazapine, which enhances both 5-HT and NA transmissions, supposedly through the antagonizing of alpha(2) adrenoreceptors, dose-dependently reduced immobility and increased climbing behaviour. Interestingly, a (mirtazapine+fluoxetine) combination treatment resulted in additive anti-immobility effects and in the summation of fluoxetine induced swimming with mirtazapine-induced climbing. Taken together, these data suggest that the NA system mediates presynaptic inhibiting interactions on the 5 HT system, that may involve alpha(2)-receptors, and that may limit the efficacy of mixed serotonin/noradrenaline reuptake inhibition in subacute antidepressant treatments. PMID- 11872335 TI - 2-Chloroadenosine, a preferential agonist of adenosine A1 receptors, enhances the anticonvulsant activity of carbamazepine and clonazepam in mice. AB - 2-Chloroadenosine (0.25-1 mg/kg) significantly raised the threshold for electroconvulsions in mice. This preferential adenosine A(1) receptor agonist (at 0.125 mg/kg) significantly potentiated the protective activity of carbamazepine against maximal electroshock-induced seizures in mice. 2-Chloroadenosine (1 mg/kg) showed also anticonvulsive efficacy against pentylenetetrazol-evoked seizures, raising the CD(50) value for pentylenetetrazol from 77.2 to 93.7 mg/kg. The drug (at 0.5 mg/kg) significantly enhanced the protective action of clonazepam in this test, decreasing its ED(50) value from 0.033 to 0.011 mg/kg. Moreover, aminophylline, a non-selective adenosine receptor antagonist (5 mg/kg), and 8-cyclopentyl-1,3-dimethylxanthine (8-CPX), a selective A(1) adenosine receptor antagonist (5 mg/kg) reversed the 2-chloroadenosine (0.125 mg/kg) induced enhancement of the protective activity of carbamazepine and clonazepam. 2 Chloroadenosine administered alone or combined with antiepileptic drugs, caused neither motor nor long-term memory impairment. Finally, the adenosine A(1) agonist did not change the free plasma concentration of antiepileptics, so a pharmacokinetic factor is not probable. Summing up, 2-chloroadenosine potentiated the protective activity of both carbamazepine and clonazepam, which seems to be associated with the enhancement of purinergic transmission mediated through adenosine A(1) receptors. PMID- 11872336 TI - A career in paediatric oncology. PMID- 11872337 TI - Current perspectives in muscle invasive bladder cancer. AB - Muscle-infiltrating bladder cancer should be dealt with in a multimodality approach with collaboration between the urologist, medical oncologist and radiotherapist. Neo-adjuvant chemotherapy has not been proven to improve survival, but may be useful in programs of bladder preservation. Response to M VAC neo-adjuvant chemotherapy is an important prognostic factor, but may represent patient selection factors. It is not known whether it is better to administer chemotherapy in the neo-adjuvant or in the adjuvant setting, that may spare some patients unnecessary chemotherapy. The international adjuvant chemotherapy trial coordinated by the EORTC (protocol 30994) will hopefully clarify some of the unanswered questions concerning whether or not adjuvant chemotherapy immediately following cystectomy improves survival. PMID- 11872338 TI - Treatment of paediatric Hodgkin's disease. a balance of risks. AB - Hodgkin's disease is one of the commoner malignancies presenting in adolescence and young adulthood and is curable in the majority of cases. A number of therapeutic regimens have been used successfully, often at the expense of the development of side-effects in later life, including second malignancies, infertility and cardiac disease. We discuss the challenge faced by paediatric oncologists today in finding the balance between maximising cure and minimising the late effects. PMID- 11872339 TI - Efficacy, tolerability and management of raltitrexed (Tomudex) monotherapy in patients with advanced colorectal cancer. a review of phase II/III trials. AB - Raltitrexed (Tomudex), a thymidylate synthase inhibitor, is an alternative to 5 fluorouracil (5-FU)/leucovorin (LV) for the first-line treatment of advanced colorectal cancer. Following the completion of four phase III studies with raltitrexed at the recommended dose of 3.0 mg/m(2), it is opportune to review the efficacy and tolerability data of raltitrexed and suggest guidelines for appropriate patient management. Data are analysed from four phase III and five phase II studies including over 1300 patients with advanced colorectal cancer, some of whom were elderly or received higher doses of raltitrexed. Median survival with raltitrexed was comparable to that of bolus or infusional 5-FU/LV in three of the four randomised studies and objective response rates in the four trials were similar for the two agents. Response rates were at least comparable in elderly patients in phase II studies. For the majority of patients, treatment with raltitrexed was well tolerated even at doses higher than that recommended or in the elderly. As with other cytotoxic agents, serious and potentially life threatening side-effects can occur; nevertheless, adherence to simple patient guidelines should minimise the incidence of serious side-effects with raltitrexed; these include the assessment of renal function before each and every treatment, dosage adjustment in the presence of renal impairment and close monitoring with prompt treatment of toxicities, particularly diarrhoea and neutropenia. PMID- 11872340 TI - Symptom response after palliative radiotherapy for patients with brain metastases. AB - Whole brain radiotherapy (RT) is frequently used to palliate symptoms in patients with brain metastases, but the palliative benefit to patients has not been well documented. We conducted a longitudinal observational prospective study of patients receiving standard RT (20 Gray (Gy)/5 fractions) for symptomatic brain metastases. End-points were observer rating of neurological symptoms, patient rated symptoms, performance status, neurological functional status, cognitive function and quality of life (QOL). Median survival for the 75 patients was 86 days (95% confidence interval (CI): 65-101 days). At 1 month, 19% of patients showed an improvement or resolution of presenting symptoms, 23% were stable and 55% had progressed or died. Patient-rated symptoms were increased at 1 month in comparison to baseline data. Only 4 patients had an improved performance status and 22 were stable. Many patients with brain metastases have a short life expectancy and may not benefit from even short duration radiation schedules. Further effort is needed to optimise patient selection and tailor treatment appropriately. PMID- 11872341 TI - Phase I pharmacokinetic and sequence finding study of the combination of docetaxel and methotrexate in patients with solid tumours. AB - This phase I study was performed to assess the feasibility and possible enhanced antitumour activity of the sequential administration of methotrexate (MTX) and docetaxel (D) in patients with solid tumours. Pharmacokinetic analysis was performed to investigate the pharmacokinetic interaction of the two agents. A total of 22 patients were enrolled, a total of six dose levels were investigated. MTX (days 1+15) 30, 40 and 50 mg/m(2)+D (day 2 or day 1) 75 and 85 mg/m(2) with supportive care measures. Both haematological and non-haematological toxicities were significant, preventing dose escalation above MTX 40 mg/m(2)+D 75 mg/m(2). Four partial responses were documented, three in patients with breast cancer, one in a patient with urothelial cell cancer. Pharmacokinetic data did not give an explanation for the significant toxicity as they revealed no interaction of D and MTX kinetics. Methotrexate and 7-OH MTX kinetics seemed to be independent of the administration of D and the moment of D administration appeared not to influence MTX kinetics. The sequential administration of MTX and D results in significant toxicity without any evidence of a clinical benefit. PMID- 11872342 TI - Effect of preoperative radiotherapy on matrilysin gene expression in rectal cancer. AB - Matrilysin, a member of matrix metalloproteinase family, is believed to play a significant role in the growth and proliferation of colon cancer cells. Overexpression of the matrilysin gene has been shown to correlate with Dukes' stage and increased metastatic potential in colorectal cancer. The aim of this study was to evaluate the effect of preoperative high-dose radiotherapy (25 Gy in five fractions over 5 days) on matrilysin (MMP-7) gene expression, in patients with resectable rectal cancer, by a quantitative reverse transcriptase-polymerase chain reaction (RT-PCR). Biopsy samples of tumour (n=30) and distant normal mucosa (n=12) from 15 patients were obtained pre- and post-radiotherapy. Messenger (m)RNA was extracted from all of the tissue samples and reverse transcribed to double-stranded cDNA. Quantitative RT-PCR was performed to study the effect of preoperative radiotherapy on matrilysin gene expression in both the tumour and normal mucosal specimens. Matrilysin mRNA values were expressed relative to glyceraldehyde-3-phosphate dehydrogenase (GAPDH) for each sample. In 14 out of 15 cases, matrilysin mRNA was detected in the cancerous tissue. Although all six normal mucosal specimens expressed matrilysin mRNA, the levels were approximately 10-fold lower compared with those seen in the paired tumour samples. Preoperative radiotherapy led to a significant 6- to 7-fold increase (P=0.001) in the expression of matrilysin mRNA in rectal cancer tissue. In contrast, there was no significant change in the matrilysin mRNA expression of normal mucosal specimens post-radiotherapy. Preoperative high-dose radiotherapy upregulates matrilysin gene expression in rectal cancer. Matrilysin inhibition may be a useful preventive or therapeutic adjunct to radiotherapy in rectal cancer. PMID- 11872343 TI - Prognostic factors of cutaneous melanoma and a new staging system proposed by the American Joint Committee on Cancer (AJCC): validation in a cohort of 1284 patients. AB - This study, involving a cohort of 1284 evaluable patients, validates the American Joint Committee on Cancer (AJCC) proposal for the introduction of ulceration of primary cutaneous melanoma as an independent prognostic factor of survival. In univariate analyses, ulceration (Hazard Ratio (HR) 1.983; P<0.0001; 95% Confidence Intervals (CI) 1.692-2.325) was a predictor of worse overall survival. In multivariate analyses, ulceration (HR 1.302; P=0.022; (95% CI: 1.039-1.633) retained its prognostic significance for survival independent of tumour thickness (HR 1.101; P<0.0001; 95% CI: 1.055-1.150); mitotic activity (HR 1.039; P=0.005; 95% CI: 1.012-1.067); and age (HR 1.009; P=0.006; 95% CI: 1.003-1.016). Ulceration lost its significance in a subgroup analysis of 256 patients with clinically apparent regional lymph node metastases to the number of lymph nodes involved (HR 1.15; P=0.004; 95% CI:1.047-1.263). Ulceration is prognostically significant in the tumour but not the nodal classification of melanoma, with mitotic activity the second most important prognostic factor after tumour thickness. PMID- 11872344 TI - Testing a new staging system for cutaneous melanoma proposed by the American Joint Committee on Cancer. AB - The American Joint Committee on Cancer (AJCC) recently proposed a new staging system for cutaneous melanoma. We tested its practicability and its prognostic value was compared with the currently used TNM classification. The data of 1976 melanoma patients were used for the testing. 1218 patients (61.6%) could be assigned to the proposed pT classification, 136 patients (90.1%) with lymph node metastases and/or in-transit metastases to the proposed pN classification and all 14 patients with distant metastases to the proposed pM classification. Proposed pathological staging was possible for 971 patients (49%). The number of pT1 patients (399 versus 230) and stage I patients (544 versus 393) was distinctly higher in the proposed classification. In proposed stage II and III groups, subgroups with different prognosis could be identified. The new staging system includes more detailed information on clinical and pathohistological findings. Nevertheless, it is practicable and enables more patients with excellent prognosis to be identified. PMID- 11872345 TI - Predictive value of thymidylate synthase expression in resected metastases of colorectal cancer. AB - Recent investigations have focused on the prognostic value of thymidylate synthase (TS) assessment in metastases of colorectal carcinoma (CRC). In order to evaluate the prognostic impact of TS expression after resection of metastases of colorectal cancer followed by systemic adjuvant chemotherapy, we performed an immunohistochemical characterisation of TS in the primary tumours and in the corresponding radically resected hepatic and pulmonary metastases. An additional objective was to compare the levels of TS in primary and metastatic disease. TS expression was assessed by immunohistochemistry using the monoclonal antibody TS 106. The study population consisted of 60 patients: 48 underwent liver and 12 lung resection. All of them received adjuvant chemotherapy after metastasectomy according to the Mayo Clinic schedule. In the 49 evaluable primary tumours, TS score was high in 53% and low in 47% of patients, while in the 60 metastatic samples TS immunostaining was high in 33% and low in 67%. There was a significantly smaller number of high TS expressors in metastatic than in primary tumours (P<0.04). No correlation was observed between TS expression and the site of the metastasis. TS status did not significantly correlate with the median disease-free interval (DFI) after metastasectomy, although this parameter was longer for patients with low TS immunoreactivity in the resected metastases than for those with high TS lesions (19.6 versus 13.8 months). Patients with high TS levels, however, had a significantly shorter median overall survival (OS) (27.6 months) than those with low TS expression (36.3 months) (P<0.008). TS status in the resected metastases confirmed its independent prognostic value in the multivariate analysis and was the only prognostic marker of OS in the subgroup of patients with resected liver metastases. These results suggest that high TS levels in resected metastases of colorectal cancer are associated with a poor outcome after surgery and 5-FU adjuvant therapy; therefore, a prospective assessment of TS levels in resected colorectal metastases could be useful to define which patients will most likely benefit from 5-FU adjuvant therapy after metastasectomy. Chemotherapeutic agents that target TS may not be the appropriate adjuvant treatment after metastasectomy for patients with a high TS expression in the resected metastases of colorectal cancer. PMID- 11872346 TI - C-erbB-2 expression does not predict response to docetaxel or sequential methotrexate and 5-fluorouracil in advanced breast cancer. AB - Breast cancer patients with c-erbB-2-positive tumours seem to benefit from anthracycline-based adjuvant chemotherapy. The predictive value of c-erbB-2 for taxane sensitivity is not yet clear. The purpose of this study was to assess whether c-erbB-2 expression is associated with clinical sensitivity to docetaxel (T) or sequential methotrexate and 5-fluorouracil (MF). A total of 283 patients with metastatic breast cancer were initially enrolled in a randomised multicentre trial comparing docetaxel with sequential MF in advanced breast cancer. Paraffin embedded blocks of the primary tumour were available for 131 patients (46%). c erbB-2 status was determined by immunohistochemistry using a polyclonal antibody to the c-erbB-2 protein. C-erbB-2 expression was scored in a semi-quantitative fashion using a 0 to 3+ scale. Staining scores 2+ or greater were considered positive. Response evaluation was performed according to World Health Organization (WHO) recommendations. Overall 54 (42%) patients had c-erbB-2 positive tumours. There was no association between treatment outcome and c-erbB-2 overexpression. The overall response rates (RR) (n=128) among c-erbB-2-negative and -positive patients were 35 and 44%, respectively (P=0.359). In the MF arm (n=62), the RR was somewhat higher in the c-erbB-2 overexpressors (33% versus 18%, P=0.18). In the docetaxel arm the RRs were very similar, regardless of the c erbB-2 expression (53% versus 53%). While several studies have suggested a prognostic and putative predictive significance of c-erbB-2 overexpression in early breast cancer, the significance of c-erbB-2 expression as a predictive factor for response to various cytotoxic treatments in advanced breast cancer is still controversial. In this study, c-erbB-2 expression could not predict response to either MF or T. Thus, tumours over-expressing c-erbB-2 are not uniformly more sensitive to taxanes and c-erbB-2 expression cannot yet be applied clinically as a predictive factor for response in advanced breast cancer. PMID- 11872347 TI - Progression-free rate as the principal end-point for phase II trials in soft tissue sarcomas. AB - We have estimated progression-free rates (PFR) for various groups of soft-tissue sarcoma patients from our clinical trials database, to provide reference values for conducting phase II studies with PFR as the principal end-point. In 146 pretreated patients receiving an active agent, the PFR estimates were 39 and 14% at 3 and 6 months; with inactive regimens (234 patients), those estimates were 21 and 8% respectively. In 1154-non-pretreated patients, PFR estimates varied from 77% (synovial sarcoma) to 57% (malignant fibrous histiocytoma (MFH)) at 3 months, and from 56% (synovial sarcoma) to 38% (MFH) at 6 months. In 61 leiomyosarcomas from gastrointestinal origin, the corresponding figures were 44 and 30%, respectively. Consequently, for first-line therapy, a 6-month PFR of > or = 30 56% (depending on histology) can be considered as a reference value to suggest drug activity; for second-line therapy, a 3-month PFR of > or = 40% would suggest a drug activity, and < or = 20% would suggest inactivity. PMID- 11872348 TI - Diagnostic strategies in non-palpable breast lesions. AB - The number of non-palpable breast lesions is growing. Needle-localised breast biopsy (NLBB) is the gold standard for evaluating these lesions. Cost-saving techniques and less invasive alternatives such as core-needle biopsy (LCNB) and fine-needle aspiration (FNA) have emerged. The aim of this study was to find out if the lesions of patients who were sent directly for surgery to undergo a NLBB differed from lesions of patients who were send for a non-operative procedure. Furthermore, if a benign result was obtained, we assessed the total and kind of subsequent diagnostic procedures that were undertaken. A retrospective study on 718 women with 749 non-palpable breast lesions was performed. In 58% of women with non-palpable breast lesion, a non-surgical procedure was chosen. Lesions sent directly for surgery were more frequently not visible on ultrasound (62%) and mainly consisted of microcalcifications only (56%). In 45%, this primary surgical approach could have been avoided. If the non-operative procedure showed a non-malignant result, 41% of these women received an additional surgical diagnostic procedure. These figures obtained from routine daily practice show the importance of protocols in order to standardise diagnostic procedures and prevent unnecessary surgery. PMID- 11872349 TI - Gemcitabine in advanced adult soft-tissue sarcomas. A phase II study of the EORTC Soft Tissue and Bone Sarcoma Group. AB - Gemcitabine (2'-deoxy-2'-difluorocytidine monohydrochloride) at a dose of 1250 mg/m(2) was given as a 30-min intravenous (i.v.) infusion on days 1 and 8 in a 3 weekly schedule to 32 patients with advanced soft-tissue sarcoma (STS) failing first-line chemotherapy. One patient was ineligible due to a delay between the previous chemotherapy and the start of treatment. Of the eligible patients, median age was 53 years (range 23-73 years). The predominant histological subtype was leiomyosarcoma in 12 patients (38%). The median number of cycles was three (range 1-8 cycles) with a median total dose of gemcitabine of 6.25 g/m(2) (range 1.25-19.97 g/m(2)). The relative dose intensity of gemcitabine was 96% (range 50 103%). Treatment was tolerated very well with non-complicated haematological toxicity as the most frequently observed side-effect. Only one partial tumour response was documented, giving a response rate of 3.23% (95% Confidence Interval (CI): 0.08-16.2%). The median overall survival was 268 days (95% CI: 129-377) and the median time to progression was 45 days (95% CI: 41-79). These results indicate that gemcitabine given at this dose and schedule is not active as second line therapy in advanced STS. PMID- 11872350 TI - Non-metastatic Ewing's sarcoma of the ribs: the French Society of Pediatric Oncology Experience. AB - From 1984 to 1997, 57 consecutive patients with non-metastatic Ewing's sarcoma of the ribs were treated according to multimodal French Society of Pediatric Oncology (SFOP) protocols EW 84, EW 88 and EW 93. The results of treatment were reviewed and analysed. Median age was 12 years. 34 patients had large tumours (greatest tumour dimension > or = 8 cm); pleural effusion was noted in 26. A tumour-positive margin after surgery was noted in 15 patients. Histological response after chemotherapy was assessed in 34 patients. 34 patients received radiation therapy. With a median follow-up of 5 years, the projected overall and relapse-free survival rates were 69 and 62%, respectively. The major site of relapse was local. None of the following was significant in predicting relapse: tumour size, gender, age at diagnosis, existence of pleural effusion, level of rib tumour, rib component, type of local control, surgical margin (positive or negative). Response to chemotherapy was the sole significant prognostic factor (P=0.004). Patients with pleural effusion had a higher percentage of relapse if they were treated without local radiation therapy. Our study confirms the prognostic significance of response to initial chemotherapy. Radiation therapy may be withheld in selected cases, but seems necessary in patients with pleural effusion. PMID- 11872351 TI - Assessment of the early impact of the population-based breast cancer screening programme in Florence (Italy) using mortality and surrogate measures. AB - The aim of this study was to evaluate the effects by the end of 1999 of the Florence breast screening programme that started in 1990. Approximately 60000 women (aged 50-69 years) were enrolled from 1990 to 1993. Breast cancer cases diagnosed from 1990 to 1996 were partitioned by the method of detection, classified by their tumour size and nodal status and followed-up for mortality at on the 31 December 1999. Incidence-based mortality in the 50-74-year-old women and advanced carcinomas rates were assessed. Due to low compliance (approximately 60%) and the long enrollment phase, only approximately 35% of the total age specific population person-years were screened. The number of invasive cases diagnosed was 1122, 17% higher than the 958 expected. After the prevalence screening, a reduction of approximately a quarter in advanced carcinomas was observed in the invited women (Odds Ratio (OR): 0.74; 95% Confidence Interval (CI): 0.55-0.98). In the period 1990-1999, 547 breast cancer deaths were observed: 78 (14%) occurred in women invited and half of these in never responders, 385 (70%) occurred in cases diagnosed before screening started. Disproportionate numbers of deaths occurred in women with advanced tumours. The 19% mortality reduction for the invited women was of borderline statistical significance (observed/expected (O/E) deaths: 0.81; 95% CI: 0.64-1.01); by a one sided test the result would be unequivocally significant. The mortality reduction attributable to screening in the whole population over the 10-year period was 3.2%. The incidence-based mortality analysis confirmed the current follow-up time is too short for screening to have had a major effect on the breast cancer mortality trends. Screening performance might be improved by a higher level of compliance and shorter interval times, but the estimate of the mortality reduction for the invited and the lower rate of advanced carcinomas confirmed that the effect of the programme is in the expected direction. PMID- 11872352 TI - Incidence of breast cancer on Crete, 1994-1995. AB - The aim of this study was to present epidemiological results relating to malignant neoplasms of breast using primary data from the island of Crete, Greece, 1994-1995. The patients were all female residents of Crete with breast cancer first diagnosed during 1994 and 1995, 208 and 207 new incident cases, respectively. The data were collected and analysed by the Cancer Registry of Crete (CRC). Direct age-standardised rates (ASR) for incidence and cumulative risk (to age 75 years) were calculated for Crete as a whole. Standardised incidence ratios (SIR) were calculated for the 20 provinces (administrative regions); these were also smoothed using Bayesian methods. The ASR for incidence per 100000 person-years was 70.6. The truncated rate (age 40 years and above) was 153.7. The SIR for the 20 provinces showed no marked variations, with three exceptions, two of which had ratios higher than 1 and one lower. Bayesian smoothing of provincial incidence rates showed that throughout Crete, the risk of breast cancer shows considerable uniformity. The incidence rate of breast cancer on Crete is higher than that of Greece overall, and is comparable with other southern European countries. A possible explanation is that the published incidence for Greece may be an underestimation of the true rate. The small variability in breast cancer incidence within Crete probably reflects the homogeneity of the population and environmental and social conditions. PMID- 11872353 TI - Primary breast cancer therapy in six regions of Germany. AB - Studies from six regions of Germany (Aachen (W1), Dresden (E1), Jena (E2), Marburg (W2), Munich (W3), and Stuttgart (C1)) have been compared to verify and assess the quality of healthcare using breast cancer as an example. All of the data collection was carried out in comprehensive cancer centres and is population based, with the exception of C1. Classic prognostic factors and the initial treatment of 8661 women with breast cancer, diagnosed between 1996 and 1998, were examined. Primary therapy, breast conserving therapy (BCT), and the use of subsequent local radiation and/or systemic therapy (chemotherapy or hormonal therapy) were analysed. BCT was performed on 39.3-57.7% of patients. By pT category, the proportion of BCT in the six regions were as follows: for pTis between 37.8 and 64.3%, for pT1 between 51.7 and 71.5%, for pT2 between 25.9 and 51.1%, for pT3 between 0 and 13.1% and for pT4 between 0 and 15.2%. Multivariate analyses, adjusted for age and biological factors, showed a significant influence of the treating hospital on the mastectomy rate. The use of radiotherapy after BCT (80%) was quite homogeneous in the six regions. The application of radiotherapy after mastectomy, however, varied between 10.4 and 32.2%. In all regions, for premenopausal patients, the use of adjuvant systemic therapy almost reflected the St. Gallen-Consensus recommendations. In contrast, post-menopausal women with positive lymph nodes were not always treated according to these standards. In all regions, age had an influence on the administration of treatment: elderly breast cancer patients received less BCT, less radiotherapy and less adjuvant therapy than recommended in the St. Gallen-Consensus. Feedback of the results was made available to each hospital, providing a comparative summary of patient care that could be used by the participating hospitals for self-assessment and quality-control. PMID- 11872354 TI - Decreased telomerase activity is not a reliable indicator of chemosensitivity in testicular cancer cell lines. AB - Telomere stabilisation is a critical step in tumorigenesis and telomerase, an enzyme which counteracts telomeric DNA loss, is active in most tumours. Conflicting evidence has been published concerning the potential use of telomerase activity as a measurement of drug-induced tumour cell killing. In this study, the time courses of telomerase loss and induction of apoptosis were investigated in two testicular cell lines, Susa CP and 833 K, following 4-h exposure to cisplatin, melphalan or doxorubicin. Telomerase activity was only affected in both cell lines at 20 h following exposure to high concentrations of cisplatin (100x the drug concentrations causing 50% growth inhibition (IC(50) values)). The time course of melphalan-induced telomerase loss, which was again only apparent at 100x IC(50) concentrations, varied between the cell lines and doxorubicin (100x IC(50)) did not induce telomerase loss in either of the cell lines. Importantly, the levels and rates of appearance of apoptotic cells (nuclear morphology and annexin V staining) were similar for all three drugs in both cell lines; i.e. cisplatin, melphalan and doxorubicin (100x IC(50)) caused similar frequencies of apoptosis in Susa CP cells at 24 h whereas telomerase activities were 65, 123 and 96% of the control, respectively. The possibility that telomerase activity was lost following cisplatin treatment through a direct interaction of cisplatin with telomerase was discounted. Additionally, the relative levels of the RNA component of telomerase (hTR) and mRNA for the telomerase catalytic subunit (hTERT) were not related to the observed decreases in telomerase activity. These data indicate that telomerase activity is not a reliable indicator of chemosensitivity in human testicular cancer cells. Furthermore, cisplatin-induced loss of telomerase activity is not due to a direct reaction with the enzyme or decreased hTR levels. PMID- 11872355 TI - Therapeutic outcome and side-effects after radiotherapy, chemotherapy and/or hyperthermia treatment of head and neck tumour xenografts. AB - The aim of the study was to optimise the still unsatisfactory therapeutic results in head and neck cancer by studying the results and the side-effects of radiotherapy, chemotherapy and/or local hyperthermia treatment of human tumour xenografts. Mice carrying human-derived head and neck squamous cell carcinoma xenografts with a mean volume of 100 mm(3) received 5x2 Gy, cisplatin or ifosfamide and/or local hyperthermia at 41/41.8 degrees C. Haematocrit and tumour volumes were determined two or three times per week, respectively, until day 25 or day 60. At day 60, the highest number of complete remissions (CRs) (80%) was observed in the triple modality therapy group with radiation, local hyperthermia at 41.8 C and cisplatin at a dosage of 2 mg/kg body weight (b.w.). Therapeutic side-effects were moderate weight loss and a mild anaemia. Thus, with regard to the long-term tumour-free survival, the most effective treatment was the combination of radiotherapy, cisplatin and local hyperthermia at 41.8 C. PMID- 11872356 TI - HPMA-hydrogels result in prolonged delivery of anticancer drugs and are a promising tool for the treatment of sensitive and multidrug resistant leukaemia. AB - Treatment of an established BCL1 leukaemia in mice showed that the use of hydrogels is advantageous in comparison with free doxorubicin (DOX), partially due to the different pharmacokinetic profile of the drug release. Pharmacologically active concentrations ranging from 100 to 800 ng/ml were detectable in the bloodstream for more than 4 days when DOX-loaded hydrogels were implanted into mice. Animals treated with free DOX survived for 35 days, survival of hydrogel-DOX treated animals increased up to 60 days and long-term survivors were achieved, when the second hydrogel was implanted 2 weeks after the first one. Hydrogels containing vinblastine (VLB) were ineffective. N-(2 hydroxypropyl)methacrylamide (HPMA) hydrogels were also used in combined therapy against multidrug resistant leukaemia P388-MDR to achieve a synergistic effect of both the cytostatic drug and chemosensitising agent. It was shown that when 4 times the maximal tolerated dose (MTD) of free DOX was incorporated into HPMA hydrogels, tumour volume was reduced by approximately 50% after implantation of the hydrogel containing DOX and cyclosporine A (CsA) and survival was slightly prolonged. PMID- 11872357 TI - Proceedings of the International Symposium on Insulin Resistance and Atherosclerosis. Osaka, Japan, 27-29 July 2000. PMID- 11872358 TI - The family of protein kinase C and membrane lipid mediators. PMID- 11872359 TI - Macrovascular disease risk factors and insulin resistance in Aboriginal and Torres Strait Islander people. AB - It has been proposed that insulin resistance (IR) underlies a cluster of cardiovascular disease (CVD) risk factors constituting a "metabolic syndrome." CVD is a leading cause of premature mortality among indigenous Australians. In a group of younger (15-44 years, fasting glucose <7.8 mmol/l) Aboriginal (n=643) and Torres Strait Islander (n=220) people participating in community-based risk factor surveys, we identified high prevalences of metabolic syndrome components: glucose intolerance, dyslipidaemia, hypertension, and IR. There were inconsistent associations of IR with other risk factors, and the data do not support a direct causal relationship between insulin and other metabolic variables. Rather, metabolic syndrome components may arise from social and environmental factors interacting with behavioural and biochemical factors in individuals. PMID- 11872360 TI - Molecular mechanism of vascular disease in metabolic syndrome X. PMID- 11872361 TI - Multifactorial insulin resistance and clinical impact in hypertension and cardiovascular diseases. AB - Insulin resistance and hyperinsulinemia have been observed in over 70% of the nonobese, nondiabetic subjects with essential hypertension (HT). Alpha-1 blockers, ACE-antagonists, long-acting Ca blockers including nifedipine CR, some form of beta-blockers, tilisolor, which is reported to increase blood flow, improve insulin sensitivity when blood pressure is better controlled. Decrease of serum potassium during insulin sensitivity test and intraplatelet free Ca2+ concentration is positively and negatively correlated with insulin sensitivity, respectively. Blood pressure is correlated with insulin resistance, which is also observed in secondary HT. The resistance is correlated with salt sensitivity as well as impaired nocturnal fall of blood pressure. These suggest the possible association of insulin resistance with altered intracellular cation metabolism. Insulin resistance and associated hyperinsulinemia have been observed in effort as well as vasospastic angina pectoris (VSAP), atherothrombotic cerebral infarction, and in ASO without obesity, HT, or diabetes, suggesting the resistance resulting from endothelial dysfunction. Insulin resistance has been observed in heart failure and is correlated with angiotensin II. Resistance is also observed in hypertrophic cardiomyopathy and is partially correlated with TNF alpha. These results indicate that insulin resistance seem to be multifactorial. An effort to normalize insulin sensitivity is crucial to eliminate multiple risk factors as well as to prevent the progression of atherosclerotic vascular lesions. PMID- 11872362 TI - The mechanism of HDL lowering in hypertriglyceridemic, insulin-resistant states. AB - Hypertriglyceridemia and reduced plasma levels of high-density lipoprotein cholesterol (HDL-c) are the most frequent forms of dyslipidemia observed in insulin-resistant states, such as obesity, impaired fasting glucose, and Type 2 diabetes, and are highly atherogenic in these settings. The hypertriglyceridemia of insulin resistance is primarily due to an overproduction of very low-density lipoproteins (VLDL), and in some instances, is also due to reduced VLDL clearance and postprandial accumulation of VLDL, chylomicrons, and their remnants [i.e., triglyceride (TG)-rich lipoproteins]. TG-rich lipoproteins actively exchange their core lipids with HDL in vivo, a process that is facilitated by cholesteryl ester (CE) transfer protein (CETP), and in hypertriglyceridemic states, this process is enhanced. This results in TG enrichment of HDL in hypertriglyceridemic states. There is accumulating evidence that TG enrichment of HDL plays an important role in determining the rate at which HDL particles are cleared from the circulation. Here, we review the evidence that TG-enriched HDL, when modulated by lipolytic enzymes in the circulation, are catabolized more rapidly than native HDL, and may ultimately explain the lowering of HDL-c in insulin resistant, hypertriglyceridemic states. Since we have recently reviewed in detail the evidence by Lamarche et al. [Clin. Chim. Acta 286 (1999) 145; J. Clin. Invest. 103 (8) (1999) 1191.] to support this hypothesis, in the present brief review, we will focus predominantly on our own recent research in this area. PMID- 11872363 TI - Atherogenic lipoproteins and diabetes mellitus. AB - The most common lipoprotein abnormality in type 2 diabetics is hypertriglyceridemia, which is known to be an independent risk factor for coronary artery disease (CAD) in diabetics. It is known that remnant lipoproteins, small, dense LDL, and isolated hypo-alphalipoproteinemia exist in diabetics even if they are apparently normolipidemic. Our previous observation revealed that type 2 diabetics had smaller LDL even if they were without hyperlipidemia. We also found that diabetics with microalbuminuria had smaller LDL than those with normoalbuminuria, indicating early nephrotoxicity of small, dense LDL. More than half of the Japanese type 2 diabetics associated with acute myocardial infarction (AMI) showed isolated hypo-alpha lipoproteinemia, indicating the clinical importance of suppressed HDL fraction without prominent hyperlipidemia in the diabetics. Finally, strict diet control and treatment of diabetics with dyslipidemias by acarbose, troglitazone, fibrates and/or statins were all successful in increasing LDL size. PMID- 11872364 TI - Influence of insulin and of insulin resistance on platelet and vascular smooth muscle cell function. AB - In this short review, we present the main results obtained in our laboratory in the last 15 years concerning the influence exerted by insulin on platelets and human vascular smooth muscle cells (VSMCs). In particular, we discuss: (i) the insulin ability to rapidly activate a constitutive nitric oxide synthase (NOS) in both cell types, with a consequent increase of the two nucleotides guanosine 3',5'-cyclic monophosphate (cGMP) and adenosine-3',5'-cyclic monophosphate (cAMP), well-known mediators of antiaggregation and vasodilation; (ii) the interplay of insulin with substances able to activate adenylate cyclase in both cell types; (iii) the impairment of the antiaggregating insulin effects in insulin-resistant subjects; (iv) the insulin-induced increase on endothelin in the VSMCs; (v) the insulin ability to slightly stimulate VSMC proliferation. PMID- 11872365 TI - The role of PPARgamma in high-fat diet-induced obesity and insulin resistance. AB - It has been well demonstrated that insulin resistance plays an important role in the clustering of coronary risk factors through the progression of atherosclerosis in animal models of insulin resistance. In humans, a high-fat diet is the major cause of obesity and insulin resistance. In this study, we investigated the role of peroxisome proliferator-activated receptor gamma (PPARgamma) in high-fat diet induced-obesity and insulin resistance by gene targeting and case-control study using the common PPARgamma2 polymorphism in human subjects. Homozygous PPARgamma-deficient embryos died at 10.5-11.5 dpc due to placental dysfunction. Heterozygous PPARgamma-deficient mice were protected from the development of insulin resistance due to adipocyte hypertrophy under a high-fat diet and the phenotypes were abrogated by PPARgamma agonist treatment. Heterozygous PPARgamma-deficient mice showed overexpression and hypersecretion of leptin despite the smaller size of adipocytes and decreased fat mass, which may explain these phenotypes at least in part. This study reveals a hitherto unpredicted role for PPARgamma in high-fat diet-induced obesity due to adipocyte hypertrophy and insulin resistance, which requires both alleles of PPARgamma. A Pro12Ala polymorphism has been detected in the human PPARgamma2 gene. Since this amino acid substitution may cause a reduction in the transcriptional activity of PPARgamma, this polymorphism may be associated with decreased insulin resistance and decreased risk of Type 2 diabetes. To investigate this hypothesis, we performed a case-control study of the Pro12Ala PPARgamma2 polymorphism. In an obese group, subjects with Ala12 were more insulin sensitive than those without. The frequency of Ala12 was significantly lower in the diabetic group, suggesting that this polymorphism protects against Type 2 diabetes. These results revealed that both in mice and humans, PPARgamma is a thrifty gene mediating Type 2 diabetes. PMID- 11872366 TI - Vascular protective effects by activation of nuclear receptor PPARgamma. AB - Pharmaceutical interventions targeting proteins that regulate VSMC growth and movement are promising new approach to treat diabetes-associated cardiovascular disease. Peroxisome proliferator-activated receptor-gamma (PPARgamma) is a ligand activated transcription factor in the nuclear receptor superfamily. Thiazolidineodione (TZT) insulin sensitizers are pharmacologic ligands for PPARgamma. All of the major cells in the vasculature express PPARgamma, including endothelial cells. VSMCs, and monocytes/macrophages. PPARgamma ligands may protect the vasculature against injury by inhibiting cell growth and movement, improving endothelial function, and suppressing tissue inflammation. Antiproliferative effects of PPARgamma ligands are mediated by targeting critical cell cycle regulators, including Rb and p27(Kip1), that regulate the progression of cells from G1 phase into S phase to conduct DNA synthesis. Pharmacologic activation of PPARgamma in vascular cells may provide a novel therapeutic approach to retard diabetes-associated vascular disease. PMID- 11872367 TI - Impact of insulin resistance on neointimal tissue proliferation after coronary stent implantation. Intravascular ultrasound studies. AB - Serial intravascular ultrasound (IVUS) studies in 55 nondiabetic patients showed that neointimal tissue proliferation after stent implantation in patients with impaired glucose tolerance (IGT) was greater than that in patients with normal glucose tolerance at follow-up. Multiple regression analysis showed that the sum of insulin levels was the best predictor of the greater neointimal index at follow-up. Another group of serial IVUS studies were performed in 62 stented lesions in 52 patients with Type 2 diabetes mellitus (DM). The study patients were randomized into a troglitazone group and a control group. The neointimal tissue proliferation at follow-up in the troglitazone group was significantly smaller than that in the control group. PMID- 11872368 TI - CD36, serves as a receptor for advanced glycation endproducts (AGE). AB - Interaction of advanced glycation endproducts (AGE) with AGE receptors induces several cellular phenomena relating potentially to diabetic complications. Five AGE receptors identified so far are receptor for AGE (RAGE), 80 K-H, OST-48, galectin-3, and macrophage scavenger receptor, types I and II (SR-A) [Eur. J. Biochem. 230 (1995) 408; Nature 386 (1997) 292.]. Since SR-A is known to belong to the class A scavenger receptor family and the scavenger receptor collectively represents a family of multiligand lipoprotein receptors, it is possible that CD36 belonging to class B scavenger receptor family (SR-B) can recognize AGE proteins as a ligand. This was tested in the present study at the cellular level by using Chinese hamster ovary (CHO) cells overexpressing human CD36 (CHO-CD36 cells). 125I-AGE-bovine serum albumin (BSA) was endocytosed in a dose-dependent fashion and underwent lysosomal degradation by CHO-CD36, but not wild-type CHO cells. Endocytic uptake of 125I-AGE-BSA by these cells was inhibited 50% by oxidized low-density lipoprotein (Ox-LDL) and 60% by FA6-152, an anti-CD36 antibody inhibiting cellular binding of Ox-LDL. Our results indicate that CD36 expressed by these cells mediates endocytic uptake and subsequent intracellular degradation of AGE proteins. Since CD36 is one of the major Ox-LDL receptors and is up-regulated in macrophage- and smooth muscle cell-derived foam cells in human atherosclerotic lesions, the present results suggest that, like Ox-LDL, AGE proteins generated in situ are recognized by CD36, which might contribute to the pathogenesis of diabetic macrovascular complications. PMID- 11872369 TI - Pathophysiological role of oxidized low-density lipoprotein in plaque instability in coronary artery diseases. AB - Oxidized low-density lipoprotein (ox-LDL) is considered to play a key role in the genesis of inflammatory processes in atherosclerotic lesions. It has also been shown that LDL isolated from patients with diabetes mellitus (DM) has an enhanced susceptibility to oxidation. Recently, a sandwich ELISA method for measurement of plasma ox-LDL levels has been developed. To elucidate the role of ox-LDL in plaque instability in coronary artery disease, we measured the plasma ox-LDL levels in patients with acute myocardial infarction (AMI), unstable angina pectoris (UAP), and stable angina pectoris (SAP), and moreover assessed whether a relationship is present between plasma ox-LDL levels and DM. We also measured the plasma ox-LDL level in a patient who died of AMI, thus enabling us to study the presence of ox-LDL and CD 36, which is one of the ox-LDL receptors, in the culprit lesion. Plasma ox-LDL levels were measured in 210 patients (AMI: 70, UAP: 70, SAP: 70), and in 55 control subjects. Plasma ox-LDL levels in AMI patients were significantly higher than in UAP patients (P<.0001), SAP patients (P<.0001), or controls (P<.0001). In the UAP group, plasma ox-LDL levels in patients with DM were significantly higher than those without DM (P<.005). The autopsied patient who died of AMI revealed an increased plasma level of ox-LDL, and immunohistochemically, the culprit coronary lesion contained abundant macrophage derived foam cells, showing distinct positivity for ox-LDL and CD 36. These results strongly suggest an important role for ox-LDL in the genesis of plaque instability in human coronary atherosclerotic lesions. PMID- 11872370 TI - Hyperglycemia-induced alteration of vascular smooth muscle phenotype. AB - We have previously reported that high glucose stimulates osteopontin (OPN) expression through protein kinase C-dependent pathway, as well as the hexosamine pathway, in cultured rat aortic smooth muscle cells (SMC). The finding prompted us to study in vivo expression of OPN in diabetes mellitus. In the present study, we found by immunohistochemistry that medial layers of the carotid arteries of streptozotocin (STZ)-induced diabetic rats, as well as the forearm arteries of diabetic patients, stained positive with OPN antibodies, whereas the staining of control rats, as well as nondiabetic patients, was negative. We also found that OPN stimulated migration and enhanced platelet-derived growth factor (PDGF) mediated DNA synthesis of cultured rat aortic SMC. OPN and PDGF synergistically activated focal adhesion kinase (FAK), as well as extracellular signal-regulated kinase (ERK), which seems to be a reason for OPN-induced enhancement of PDGF mediated DNA synthesis. Taken together, our present results raise a possibility that OPN plays a role in the development of diabetic vascular complications. PMID- 11872371 TI - Role of glycogen synthase kinase-3 in skeletal muscle insulin resistance in Type 2 diabetes. PMID- 11872373 TI - Lysophosphatidylcholine inhibits the expression of prostacyclin stimulating factor in cultured vascular smooth muscle cells. AB - We have cloned a prostacyclin (PGI2) stimulating factor (PSF), which stimulates PGI2 production by vascular endothelial cells. Previous study demonstrated the reduced PSF expression in the coronary arteries from the patients with ischemic heart disease. To clarify the mechanism of reduced PSF expression in atherosclerosis, we examined the effect of lysophosphatidylcholine (lysoPC), a main component of oxidized low density lipoprotein (LDL), on PSF expression in cultured vascular smooth muscle cells. LysoPC reduced PSF expression dose dependently. Whereas neither phosphatidylcholine nor native LDL affects the PSF expression. Calphostin C, a protein kinase C (PKC) inhibitor, restored the reduction of PSF expression by lysoPC. These results suggest that lysoPC-induced reduction of PSF expression is mediated by PKC activation and is playing a role in the initiation and progression of atherosclerotic lesions. PMID- 11872374 TI - Importance of blood pressure control in patients with diabetes mellitus. PMID- 11872372 TI - Hexosamines as mediators of nutrient sensing and regulation in diabetes. AB - High concentrations of glucose induce insulin resistance, impair insulin secretion, and affect hepatic glucose production in a manner that mirrors Type 2 diabetes, and hexosamines mimic many of these effects. This has led to the hypothesis that cells use hexosamine flux as a glucose- and satiety-sensing pathway. The hexosamine hypothesis for glucose sensing has been validated by overexpressing the rate-limiting enzyme for hexosamine synthesis, glutamine: fructose-6-phosphate amidotransferase (GFA) in several tissues including muscle, liver, fat, and beta cells. With overexpression of GFA in transgenic animals, skeletal muscle becomes insulin resistant, the liver synthesizes excess fatty acid, and the beta cell secretes excess insulin leading to hyperinsulinemia. Thus, excess hexosamine flux leads to a coordinated response whereby fuel is shunted toward long-term storage, mirroring the "thrifty phenotype." Chronically, however, these same adaptive changes result ultimately in obesity, hyperlipidemia, beta cell failure, and Type 2 diabetes. These results suggest a mechanism by which chronic overnutrition leads to the phenotype of Type 2 diabetes. PMID- 11872375 TI - Insulin resistance and vascular function. AB - It has become clear that amongst its many actions insulin is also a vasoactive hormone. Its effect to cause endothelial-nitric oxide-dependent vasodilation is physiologic and dose-dependent. Recent data suggest that insulin's metabolic and vascular actions are closely linked. Indeed, insulin resistant states, which by definition, exhibit diminished insulin-mediated glucose uptake into peripheral tissues also display impaired insulin mediated vasodilation as well as impaired endothelium dependent vasolidation to the muscarinic receptor agonist acetylcholine. Free fatty acids are elevated in states of insulin resistance and also cause endothelial dysfunction along with impaired insulin-mediated vasodilation. Thus, a picture is emerging linking insulin action in peripheral tissues to its action in endothelium. More recent data suggest that insulin signaling mechanisms in peripheral tissues and endothelium may be shared. Thus mechanisms causing insulin resistance via defects in insulin signaling might be expected to be manifest in both tissues. The protective action of nitric oxide and healthy endothelial function are critical to prevent atherosclerotic vascular disease. If follows that endothelial dysfunction associated insulin resistance through common defects in insulin signaling presents a parsimonious mechanism to account for the increased risk of cardiovascular disease associated with insulin resistance. PMID- 11872376 TI - Renal endothelial dysfunction and hypertension. AB - Patients with essential hypertension have an impaired endothelium-dependent vascular relaxation in the renal arteries. The possible mechanisms by which essential hypertension is associated with alterations in endothelial function are decreased endothelial nitric oxide (NO) synthase activity, decreased availability or deficiency of L-arginine, increased endogenous NO synthase inhibitor, inactivation of NO by superoxide anions, and increased vasoconstrictors. However, the precise mechanism is not known. In addition, we are now confronted with a difficult question. And the question is whether endothelial dysfunction is a cause or consequence of hypertension. We hypothesize that the initial endothelial dysfunction raises blood pressure, and the development of hypertension impairs much more endothelial function, resulting in constituting the vicious cycle between endothelial dysfunction and hypertension. However, at the moment, it is impossible to answer this question with any certainty. Impairment of endothelial function has been shown to play a critical role in the development and maintenance of hypertension. It is clinically important to select an appropriate intervention that is effective in improving endothelial dysfunction in patients with essential hypertension. Several investigators including us have demonstrated that certain interventions improve endothelial dysfunction of forearm and renal circulation in patients with essential hypertension: angiotensin-converting enzyme inhibitors; lifestyle modification: exercise, body weight reduction, and sodium reduction; estrogen replacement in postmenopausal women; and novel properties: vitamin C and tetrahydrobiopterine. In patients with essential hypertension, endothelial function is impaired in several arteries. However, endothelial dysfunction in essential hypertension is reversible. We can restore endothelial function in essential hypertensive patients. PMID- 11872377 TI - Roles of peroxisome proliferator-activated receptor gamma in cardiovascular disease. AB - Peroxisome proliferator-activated receptors (PPARs) are transcription factors belonging to a nuclear receptor superfamily. PPARs have three isoforms: alpha, beta (or delta), and gamma. It is known that PPARgamma is expressed predominantly in adipose tissue and promotes adipocyte differentiation and glucose homeostasis. Recently, synthetic antidiabetic thiazolidinediones (TZDs) and the natural prostaglandin D2 (PGD2) metabolite, 15-deoxy-Delta(12,14)-prostaglandin J2 (15d PGJ2), have been identified as ligands for PPARgamma. Furthermore, it has become apparent that PPARs are present both in a variety of different cell types and in atherosclerotic lesions and the studies about PPARgamma have been extended. Although activation of PPARgamma appears to have protective effects on atherosclerosis, it is still largely uncertain whether PPARgamma ligands prevent the development of cardiovascular disease. Recent evidence suggests that some benefit from antidiabetic agents, TZDs, may occur independent of increased insulin sensitivity. In this article, we review the latest developments in the PPAR field and summarize the roles of PPARgamma and the actions of PPARgamma ligands in the cardiovascular system. PMID- 11872378 TI - Clinical implication of multiple risk factor control in the management of diabetic macrovasucular complications. AB - Using cross-sectional and prospective analyses, the risk factors for macroangiopathy (MA) in nonobese Type 2 diabetic patients were evaluated. In the cross-sectional study, we determined a cutoff point for each variable at which changes in the prevalence of total MA reached statistically significant levels. In the prospective study, those who met more than four out of seven control criteria as set forth in the Multiclinical Study for Diabetic Macroangiopathy (MSDM) had less risk of MA in Type 2 diabetes initially diagnosed without MA compared with those who fulfilled less than three factors. These results suggest that multiple risk factor control is the most effective and reasonable way to lower the incidence of MA in Type 2 diabetes. PMID- 11872379 TI - Pathophysiogical role of leptin in lifestyle-related diseases. Studies with transgenic skinny mice overexpressing leptin. AB - Leptin is a major adipocyte-derived hormone that is involved in the regulation of food intake and energy expenditure. Plasma leptin concentrations are elevated in obese subjects, suggesting its pathophysiological role in obesity-related lifestyle-related diseases. We have recently succeeded in the generation of transgenic skinny mice overexpressing leptin. They exhibit increased glucose metabolism and insulin sensitivity accompanied by a significant increase in insulin signaling for glucose utilization in the skeletal muscle and liver. They also show blood pressure elevation through the sympathetic activation. Introduction of the lethal yellow agouti (A(y)) allele into transgenic skinny mice results in late-onset obesity and diabetes with blood pressure elevation similar to those found in nontransgenic agouti mice (A(y)/+ mice). After caloric restriction, blood pressure elevation is reversed but insulin resistance still remains in A(y)/+ mice in parallel with a reduction of plasma leptin concentrations. By contrast, blood pressure elevation is sustained but insulin resistance is reversed in transgenic mice overexpressing leptin with the A(y) allele (Tg/+:A(y)/+ mice), which remain hyperleptinemic. Collectively, our data suggest the pathophysiologic and therapeutic implication of leptin in obesity related insulin resistance and hypertension. PMID- 11872380 TI - The pathophysiologic basis of efficacy and clinical experience with the new oral antidiabetic agents. AB - Type 2 diabetes results from the abnormal resistance of peripheral tissues to insulin and from the progressive insulin secretory failure of the pancreatic beta cells. Treatment of type 2 diabetes has greatly improved due to the availability of new classes of oral antidiabetic drugs (OADs) and new insulin analogs. Three types of oral medications exert their antidiabetic action without directly stimulating insulin release: alpha-glucosidase inhibitors (e.g., acarbose) interfere with the digestion of dietary glucose precursors and the absorption of glucose; biguanides (e.g., metformin) inhibit hepatic gluconeogenesis, thereby lowering fasting blood glucose concentrations and increasing peripheral insulin sensitivity; and thiazolidinediones (e.g., rosiglitazone) improve the sensitivity of tissues to insulin-stimulated glucose disposal. In contrast, two classes of OADs stimulate insulin release from pancreatic beta-cells. Sulfonylureas (e.g., glyburide) have been used successfully for many years to treat type 2 diabetes, but their prolonged action may result in hypoglycemia. The third-generation sulfonylurea glimepiride is associated with a reduced risk of hypoglycemia and less weight gain than other sulfonylureas. Finally, the meglitinides (e.g., repaglinide) and D-phenylalanine derivatives (e.g., nateglinide) are powerful prandial insulin secretagogues. If the pancreatic beta-cells deteriorate to such an extent that insulin secretion is significantly impaired, treatment with additional exogenous insulin may be required. PMID- 11872381 TI - Genetically manipulated vectors of human disease: a practical overview. PMID- 11872382 TI - Chasing coccidia--new tools enter the race. AB - The 8th International Coccidiosis Conference, held on 9--13 July 2001 in Palm Cove, Australia, was a showcase of the latest studies on widely known coccidia, including Eimeria and Toxoplasma in addition to the emerging or re-emerging parasites such as Neospora, Cryptosporidium and Cyclospora. This meeting was staged in conjunction with the Annual Scientific Meeting of the Australian Society for Parasitology. PMID- 11872383 TI - American Society of Parasitologists presents parasites on the Rio Grande. PMID- 11872393 TI - Spirocerca lupi and dogs: the role of nematodes in carcinogenesis. PMID- 11872384 TI - Protective saliva: sandfly spit delivers. PMID- 11872395 TI - Zooprophylaxis: are we in Plato's cave? PMID- 11872396 TI - Heterogeneities in anti-schistosome humoral responses following chemotherapy. AB - Schistosomiasis is a major public health problem in Africa, the Middle East, Asia and South America. The main control strategy is to treat infected people with anthelmintic drugs, principally the safe and relatively cheap drug praziquantel. Several treatment re-infection studies in humans have shown that praziquantel can have long-term effects beyond a transient reduction of infection intensity. These long-term effects include the altering of schistosome-specific immune responses in humans, which is associated with resistance to re-infection. Differences have been observed in treatment-induced immunological changes between individuals and between populations. This article discusses the contributions of host- and parasite-related heterogeneities to post-treatment humoral responses in humans infected with Schistosoma mansoni and Schistosoma haematobium and considers the practical implications of such heterogeneity for schistosome immuno-epidemiology studies. PMID- 11872397 TI - Evolution of gametocyte sex ratios in malaria and related apicomplexan (protozoan) parasites. AB - 'Survival of the fittest' is usually interpreted to mean that natural selection favours genes that maximize their transmission to the next generation. Here, we discuss recent applications of this principle to the study of gametocyte sex ratios in malaria and other apicomplexan parasites. Sex ratios matter because they are an important determinant of fitness and transmission success -- and hence of disease epidemiology and evolution. Moreover, inbreeding rates can be estimated from gametocyte sex ratios. The sex ratio is also an excellent model trait for testing the validity of important components of what is being marketed as 'Darwinian medicine'. PMID- 11872399 TI - Decoding the language of var genes and Plasmodium falciparum sequestration. AB - Sequestration and rosetting are key determinants of Plasmodium falciparum pathogenesis. They are mediated by a large family of variant proteins called P. falciparum erythrocyte membrane protein 1 (PfEMP1). PfEMP1 proteins are multispecific binding receptors that are transported to parasite-induced, 'knob like' binding structures at the erythrocyte surface. To evade immunity and extend infections, parasites clonally vary their expressed PfEMP1. Thus, PfEMP1 are functionally selected for binding while immune selection acts to diversify the family. Here, we describe a new way to analyse PfEMP1 sequence that provides insight into domain function and protein architecture with potential implications for malaria disease. PMID- 11872398 TI - Aspartic proteases of Plasmodium falciparum and other parasitic protozoa as drug targets. AB - All parasitic protozoa contain multiple proteases, some of which are attracting attention as drug targets. Aspartic proteases are already the targets of some clinically useful drugs (e.g. chemotherapy of HIV infection) and a variety of factors make these enzymes appealing to those seeking novel antiparasite therapies. This review provides a critical analysis of the current knowledge on Plasmodium aspartic proteases termed plasmepsins, proposes a definitive nomenclature for this group of enzymes, and compares these enzymes with aspartic proteases of humans and other parasitic protozoa. The present status of attempts to obtain specific inhibitors of the parasite enzymes that will be useful as drugs is outlined and suggestions for future research priorities are proposed. PMID- 11872400 TI - Fc receptors and immunity to parasites. AB - Fc receptors (FcRs) are crucial in the immune system; they mediate a plethora of biological functions as diverse as antigen presentation, phagocytosis, cytotoxicity, induction of inflammatory cascades and modulation of immune responses. Parasites, in order to survive in the immunocompetent host, have devised ingenious methods to subvert this important aspect of the immune response. This article discusses the current thinking on FcRs, their role in immunity to parasites, and immune evasion strategies employed by parasites in their attempt to neutralize the important immune defense mechanisms mediated by these molecules. PMID- 11872405 TI - Nail changes in genodermatoses. AB - Nail changes may be marker lesions for complex systemic disorders and herald associated syndromes. Knowledge of the anatomy, embryology and biochemical properties of the nail apparatus is essential for understanding the pathogenesis of hereditary nail disorders. In the last few years significant progress has been made in the field of clinical and molecular pathology of human diseases. A considerable number of the genes responsible for genodermatoses have been identified. The homeobox master control genes, genes encoding for transcription factors, genes encoding for the maintenance of telomeres, or for structural molecules, such as the similarly evolutionary highly conserved a-helical rod domains of keratins, are involved in the embryogenesis and normal functioning of nails. Using nail changes in selected genodermatoses with a known genetic background, we try to elucidate the genesis of inherited nail disorders and review the resultant clinical manifestations. PMID- 11872406 TI - Mal de Meleda without mutations in the ARS coding sequence. AB - Mal de Meleda (Mal de Mljet) is a recessive palmoplantar hyperkeratosis associated with hyperhidrosis, brachydactyly and sometimes pseudo-ainhum. It was recently discovered to be caused by mutations in the ARS gene on chromosome 8. Here we report a patient suffering from Mal de Meleda not associated with ARS mutations. The related E48 and GML genes were also excluded. PMID- 11872407 TI - Speckled lentiginous nevus syndrome: delineation of a new distinct neurocutaneous phenotype. AB - Speckled lentiginous nevus syndrome, a so far unrecognized cutaneous phenotype associated with neurological anomalies, is postulated on the basis of the following arguments. Phacomatosis pigmentokeratotica represents a twin spot phenomenon. One isolated half of this complex phenotype is observed rather often in the form of Schimmelpenning syndrome, whereas the other half of this twin nevus syndrome consists of a speckled lentiginous nevus associated with various neurological abnormalities incompatible with Schimmelpenning syndrome, such as hyperhidrosis, muscular weakness and dysesthesia. This second component of phacomatosis pigmentokeratotica may likewise occur separately. For the association of speckled lentiginous nevus with hyperhidrosis, muscular weakness, dysesthesia or other neurological abnormalities, the term "speckled lentiginous nevus syndrome" is proposed. Some case reports that may be categorized as examples of this new syndrome are reviewed. The postulated new phenotype is tentatively categorized as a paradominant trait. Future clinical studies will probably confirm the existence of speckled lentiginous nevus syndrome as a distinct neurocutaneous phenotype. PMID- 11872408 TI - The effect of two moisturisers on skin barrier damage in allergic contact dermatitis. AB - Nickel (Ni)-induced allergic contact dermatitis (ACD) was performed in human volunteers to study the role of moisturisers in preventing ACD-related skin barrier damage. 15 Ni-sensitive females (mean age: 29.5 years; range: 23-38) were included. On days 1, 21, 24 and 26, TEWL, stratum corneum (SC)-capacitance and clinical score were evaluated on four test sites on the right and left forearms. Both a highly and a poorly hydrating moisturising formulation were applied on two sites from days 1 to 21, after which Ni-ACD was induced on the 2 pre-treated sites and one non-treated area. On day 24, TEWL values were significantly increased on the site pre-treated with poorly hydrating product compared to the rich formulation pre-treated site. SC-hydration was significantly improved on the latter site on days 21, 23 and 26. Long-term use of inadequate moisturiser increases skin barrier damage due to Ni-ACD. PMID- 11872409 TI - Changes of comedonal cytokines and sebum secretion after UV irradiation in acne patients. AB - The aims of this study were to investigate the effect of UVA and UVB on comedones and sebum secretion in acne patients. Thirteen acne patients were irradiated by UVA (starting from 20J/cm2 and increasing 10% every day) and UVB (starting from 2/3 MED and increasing 10% every day). IL-1alpha, IL-6, IL-1 receptor antagonist (IL-1ra), IL-10 and GM-CSF were measured by ELISA. Measurement of sebum level was also performed. Sebum level was increased in the first three days by UVA (18.4 - > 37.6 microg/cm2) and UVB (19.1 --> 40.0 microg/cm2), but subsequently returned to normal values. Production of IL-1alpha, IL-1ra, IL-6, and IL-10 was generally higher on day 5 than on day 10. GM-CSF was not detected from all comedones. After UV irradiation, clinically stationary acne patients showed a higher increase in cytokine production compared with improved acne patients. It is suggested that IL 10 & IL-1ra have key roles in this cytokine network as the anti-inflammatory comedonal cytokines. They may play important roles in the immuno-regulation, which may be disturbed in stationary acne patients. PMID- 11872410 TI - Comparison of the effects of tretinoin, adapalene and collagenase in an experimental model of wound healing. AB - Adapalene is a new naphthoic acid derivative with strong retinoid agonistic pharmacological properties. We propose that adapalene might contribute to the wound repairing process as is detected with retinoids. In this controlled study, the effects of topical adapalene, tretinoin and collagenase on full-thickness wound healing were compared in an animal model. Thirty-two adult male Wistar Albino rats were used in the study. Two circular, full-thickness wounds were made for each animal with a standard 8-mm punch biopsy, on both sides of the midline on the back. No treatment was given to Group I rats (n:8) which comprised the control group. Tretinoin cream (0.1%) was applied topically in Group II (n:8), adapalene gel (0.1%) in Group III (n:8), and collagenase ointment in Group IV (n:8) once daily. On day 7, the wounds were photographed to measure the wound surface area. The wounds on the left side of each animal were excised on day 7, for histopathologic and biochemical examination. The treatments were continued for the right side wounds up to 14 days when the same procedure was repeated. In Group II, a significant decrease in hydroxylproline (HP) levels was detected at day 7 (p = 0.018), and an increase at day 14 (p = 0.002) compared to the control group. HP results revealed no difference either in Group III nor in Group IV versus control at day 7 or 14. However, findings of improved healing were more prominent in Groups II and III than the other groups in histopathologic examination. In conclusion, tretinoin and adapalene contributed to the wound healing process resulting in an enhancement of collagen production, angiogenesis and granulation tissue formation. PMID- 11872411 TI - Number and distribution of interstitial cells lining the epidermis of normal human skin from different anatomical locations. AB - A study was conducted to determine the number and distribution of interstitial cells lining the epidermis of normal human skin from various anatomical locations. Five to seven normal human skin specimens per each anatomical site were collected from surgical specimens, mostly with pigmented nevi. Ten anatomical locations were classified; and a total of 65 normal human skin samples were evaluated. The number of interstitial cells lining the epidermis was quantified under light microscopy using a computer-assisted image analyzer. Two types of interstitial cells were recognized beneath the dermo-epidermal junction of normal human skin: cells containing oval nuclei and spindle-shaped cells containing elongated nuclei. As for the number of oval cells, no significant difference was found among the anatomical locations. In contrast, significantly greater numbers of spindle-shaped cells were found in the palm (4.11 + 1.24; p < 0.01), sole (3.52 + 0.83; p < 0.001) and buttock (2.52 + 0.49; p < 0.01), compared with those in the anterior trunk (0.60 + 0.22). In normal skin of the palm and sole, the number of spindle-shaped cells located beneath the apices of rete ridges (7.35 + 1.56) was significantly greater than along the dermal papillae (1.39 + 0.39, p < 0.01). However, cells containing oval nuclei also predominated beneath the apices of rete ridges, but the difference was not significant. In summary, the present study demonstrated that the number of spindle-shaped cells, quantified by H & E staining, was significantly greater beneath the apices of rete ridges than in dermal papillae. The number was greater in palm and sole skin compared with other samples of normal human skin. This data may relate to the glabrous nature of palm and sole skin. PMID- 11872412 TI - Effects of topical petrolatum and salicylic acid on the erythemogenicity of UVB. AB - Various topical agents used in combination with phototherapy have blocking effects, however in contrast to in vitro studies there were not enough in vivo studies about this subject. Our purpose was to examine the photoprotective effects of white petrolatum and salicylic acid which can be used before UVB therapy in psoriasis patients. In 35 volunteers, a phototest was performed to determine the minimal erythema dose (MED) and the test was repeated with thin (0.1 cc/25 cm2) and thick (0.3 cc/25 cm2) petrolatum, thin and thick salicylic acid (20%) in petrolatum and sunscreen. After 24 hrs, the effects of each agent on MED was investigated. MED values detected after pure UVB and after application of topical agents were compared one by one, and the differences between all of them have been found statistically significant. These showed that MED values were increased by thin or thick petrolatum and also by thin and thick salicylic acid (20%) in petrolatum. The highest MED values were detected with thick salicylic acid in petrolatum followed by thin salicylic acid in petrolatum, thick petrolatum and thin petrolatum. The application of petrolatum and salicylic acid, which can block UVB, before phototherapy is not recommended. PMID- 11872413 TI - Treatment of tinea pedis with a single pulse of itraconazole. AB - An open-design, prospective, non-comparative study was conducted to evaluate the efficacy and safety of itraconazole in the treatment of tinea pedis (interdigital type, hyperkeratotic-type and the combination of both). Treatment consisted of one pulse of itraconazole, meaning that each patient received 400 mg/day for one week. Clinical and mycologic control examinations were performed at baseline and at the end of treatment; follow-up visits took place at 30 and 60 days after the last medication administration. A total of 44 patients were evaluated. The major causal agent isolated was Trichophyton rubrum (93%). At the final follow-up visit (60 days), 37 cases (84.4%) achieved clinical and mycologic cure; 5 (11.3%) had improvement and 2 cases (4.5%) failed. Three cases reported side effects attributable to itraconazole (6.6%); one patient had a moderate headache and two reported moderate dyspepsia. None of the 3 cases required discontinuation of the medication. We concluded that the administration of one pulse of itraconazole is an effective, safe and short regimen to treat tinea pedis. PMID- 11872415 TI - Usefulness of erythrocyte sedimentation rate as tumor marker in cancer associated dermatomyositis. AB - Dermatomyositis is an inflammatory myopathy characterized by proximal symmetrical muscle weakness with a characteristic cutaneous eruption. Population-based cohort studies provide evidence of an increased frequency of cancer in dermatomyositis patients. Many signs and serology tests have been suggested as markers for malignancy in dermatomyositis. We performed a case-control study on the patients admitted in our institutions for dermatomyositis. Clinical and laboratory data were collected and statistical analysis was performed to reveal important predictive signs of malignancy in dermatomyositis. We found no statistical difference in the clinical or laboratory parameters between the dermatomyositis patients with or without malignancy, with the exception of the erythrocyte sedimentation rate. In our study group an erythrocyte sedimentation rate higher than 35 mm/hr was very strongly associated with the presence or the development of a malignancy. Given the high positive and negative predictive values observed in our study, erythrocyte sedimentation rate evaluation using this cut-off point could be very useful in alerting dermatologists to the need for more in-depth diagnostic procedures in dermatomyositis patients. PMID- 11872414 TI - Pityriasis versicolor rubra. AB - We report six typical cases with pityriasis versicolor (PV) rubra, with a background of collagen diseases in five cases and none in one case. Two cases of systemic lupus erythematosus (SLE) and one case of systemic scleroderma (SSc) had both PV rubra and nigra on the trunk. Diagnosis of superficial infections of PV was made by microscopic examination of skin scrapings following KOH, and many small whitish colonies were obtained in Sabouraud's slant agar medium culture containing cycloheximide with olive oil in all cases. Malassezia sympodialis was isolated from the scales of two different lesions at a 6 week-interval in a same person (a 32 year-old male without SLE nor SSc) by the method of Makimura et al. [5], although the other five cases were not examined for the isolation. Histopathological features of the lesion on the dorsum of the trunk showed no epidermal hyperplasia without elongation of rete ridges and no inflammatory cell infiltration in the dermis, however there was only dilatation of small blood vessels in the dermis, which was reconfirmed capillaroscopically. In the horny layers, several yeastlike and fine filamentous structures were seen which were positive with PAS and Grocott stains. Both clinical and histological features led us to speculate PV rubra. All the patients were treated with anti-fungal ointment, and the lesions diminished in less than 2 weeks. No recurrence has been seen in any of the cases. This PV rubra may be independent from PV nigra, although Horiuchi [2] suggested the earlier lesion occurs in advance of PV alba or nigra. PMID- 11872416 TI - Prospective case-control study of chlamydia, legionella and mycoplasma infections in patients with pityriasis rosea. AB - A double-blind placebo-controlled trial reported the benefit of erythromycin in treating pityriasis rosea (PR), a postulated mechanism being the eradication of bacteria susceptible to erythromycin. The aim of this study was to investigate the association between PR and Chlamydia pneumoniae, C. trachomatis, Legionella longbeachae, L. micdadei, L. pneumophila, and Mycoplasma pneumoniae infections. We recruited 13 patients aged seven to 46 years (mean: 26.8 years) diagnosed to have PR in a primary care setting in 18 months. Lesional histopathology was arranged for atypical cases. Clotted blood was collected at initial presentation and four weeks later. Controls were 13 paired age-and-sex-matched patients requiring blood collection for non-dermatological diseases. Serology tests were performed in parallel but were read "blinded" on the acute and convalescent specimens of patients and the control subjects. The serology profiles were not diagnostic of active infection by any of the bacteria studied for all 13 patients. Two patients had four-fold increase in IgG titres against C. pneumoniae, with IgM being negative. Two patients had IgM detectable against L. pneumophila serotype 6 and M. pneumoniae respectively, with no significant rise of the specific IgG. These patients had no symptom or sign of chest infection. The seroprevalence and IgG titres of the study patients for the bacteria investigated were insignificantly different from those of control subjects. We conclude that the bacteria investigated in this study do not play a significant role in the pathogenesis of PR. We believe that anti-inflammatory and immunomodulatary effects might contribute towards the action of erythromycin, if any, in PR. PMID- 11872417 TI - Steroid sparing effect of intravenous immunoglobulin therapy in patients with pemphigus foliaceus. AB - Pemphigus foliaceus (PF) is a rare autoimmune cutaneous blistering disease, with only skin involvement. Systemic corticosteroids and immunosuppressive agents are the mainstay of therapy. However, some patients develop multiple side effects to systemic corticosteroids, when they are used in high doses over prolonged periods. In some patients, immunosuppressive agents are not effective or contraindicated. In such patients, alternative treatment modalities are needed. The purpose of this study is to demonstrate the use of intravenous immunoglobulin (IVIg) therapy in seven patients with severe PF, who were steroid-dependent. The following information was recorded in each patient, before and after IVIg therapy initiation: total dose and total duration of prednisone therapy, and number of relapses. In addition, the highest dose and side effects of prednisone therapy, and duration of observation were documented. After the initiation of IVIg treatment, doses of systemic corticosteroids were gradually reduced and eventually discontinued over a mean period of 2.8 months. Thereafter, IVIg was used as monotherapy. A statistically significant difference was noted between pre IVIg therapy and after the initiation of IVIg therapy in the total dose (p = 0.005), and total duration of prednisone treatment (p = 0.02), and the number of relapses (p = 0.002). In all seven patients, IVIg produced an effective clinical response and demonstrated a steroid-sparing effect. In patients with PF, who are steroid-dependent and in whom use of conventional immunosuppressive agents is contraindicated, IVIg appears a safe and effective agent to induce and maintain a prolonged clinical remission. PMID- 11872418 TI - Allergic contact dermatitis in hairdressers: frequency and source of sensitisation. AB - Occupational skin diseases are very common among hairdressers. The aim of our study was to evaluate the frequency, the age distribution and the source of contact sensitization in a group of 209 hairdressers who consulted our Clinic from 1990 to 1999. One hundred and thirty-two patients showed one or more clinically relevant positive reactions to different allergens; 89 of them were positive to the hairdressers' series and 43 were positive to other allergens. Para-phenylenediamine base and para-toluenediamine sulphate caused the greatest number of positive reactions (77 and 29 respectively). Both glyceryl monothioglycolate and ammonium persulphate gave 25 positive patch tests. The allergens not included in the hairdressers' series which gave the greatest number of positive reactions were nickel sulphate and disperse dyes yellow 3, blue 124 and red 1. The allergens known as strong skin sensitizers have remained almost the same over the years. Preventive measures should be mandatory to protect hands and to improve the safety of this job. PMID- 11872419 TI - Giant orf on the nose. AB - Ecthyma contagiosum, or orf, is a viral zoonosis of sheep and goats that can be transmitted to humans. In humans, it generally manifests as a solitary skin lesion, although rarely it can have an unusual course or be accompanied by systemic symptoms or complications. We present a case of giant orf lesion on the nose of a 9-year-old. The lesion grew rapidly and measured 5 cm by 4 cm and was attached to the right ala nasi by a base 2 cm round in diameter. The diagnosis was suggested by clinical and histopathologic appearance and confirmed by electron microscopic visualization of the virus. The lesion resolved spontaneously with minimal scarring and the entire cycle lasted about 3 months. PMID- 11872420 TI - Multiple sclerosis and bullous pemphigoid: a casual association or a pathogenetic correlation? AB - The literature reports about thirty cases of bullous pemphigoid (BP) which developed in patients affected by multiple sclerosis (MS), although a relationship between these two diseases has not yet been identified. We report two cases of female patients affected by MS who subsequently developed BP. Two women came to our observation because of the presence of a bullous dermatitis. One, aged 54, had been affected by rapidly evolving MS for three years while the other, aged 60, had been suffering from MS since the age of 43. The clinical suspicion of BP was confirmed by cytodiagnostic examination, histology and immunofluorescence. Parenteral steroid therapy produced a rapid improvement of the clinical picture in both cases with a relapse after one year. On the basis of the cases observed, in which BP appeared in the absence of known potential triggering causes, and following a revision of the literature, we agree with those authors who assert that the two diseases could, even though only sporadically, be correlated and we think that the neurological disease could be the cause of the manifestations through immunological pathogenetic mechanisms. PMID- 11872421 TI - HPV 29-associated spotted hyperpigmentation of the face. AB - A healthy immunocompetent 40-year-old man presented with spotted hyperpigmentation of the face. Skin-biopsies taken from these lesions revealed slight acanthosis of the epidermis with vacuolization within the keratinocytes in the upper malpighian layer similar to HPV-induced cytopathic viral effects. Polymerase chain reaction analysis and subsequent direct DNA sequencing could clearly demonstrate the presence of HPV 29 DNA in the hyperpigmented macules. To our knowledge, this is the first report of facial hyperpigmented lesions induced by HPV 29. PMID- 11872422 TI - Primary cutaneous plasmacytoma on chronic lymphoedema. AB - An unusual case of cutaneous plasmacytoma with multiple lesions in a 77-year-old man affected by chronic lymphoedema of his lower limbs is reported. Histopathology and immunohistochemical staining led to the diagnosis of cutaneous plasmacytoma. Staging studies were not able to identify extracutaneous involvement. The lesions were successfully treated with melphalan and prednisone. No signs of progression of the disease were seen at a 2- year follow-up. PMID- 11872423 TI - Panniculitis after subcutaneous injection of interferon beta in a multiple sclerosis patient. AB - BACKGROUND: Interferons are used in the immune therapy of multiple sclerosis, Kaposi's sarcoma, hepatitis and melanoma based on their antiviral, immunoregulatory and antitumor activities. We report a rare side effect observed during treatment of multiple sclerosis with subcutaneous interferon beta injections. OBSERVATIONS: A 44-year-old patient diagnosed with multiple sclerosis received immune therapy with 6 Mio IU recombinant interferon-beta (IFN-beta) every other day. After 4 years of subcutaneous interferon injections painful indurations appeared directly at and adjacent to the injection sites on the thighs, arms and abdomen. Pain in the thighs made walking almost impossible and required therapy with opiates. After changing to another interferon-beta medication the symptoms improved. Subsequent treatment with doxycycline for 3 weeks was of no additional benefit. CONCLUSIONS: Panniculitis at the injection sites is a rare event but has a significant impact on the quality of life during interferon therapy. PMID- 11872424 TI - Neutrophilic eccrine hidradenitis in actinic reticuloid syndrome. AB - Neutrophilic eccrine hidradenitis (NEH) is a rare, transient complication, which usually occurs in patients with leukemia receiving various chemotherapeutic regimens. However, similar eruptions have been observed in other conditions, including HIV-positive patients, and are often preceded by the onset of malignancies. We report the first case of NEH arising in a patient with actinic reticuloid syndrome who had been treated with methotrexate. PMID- 11872425 TI - Telangiectasia macularis eruptiva perstans in polycythemia rubra vera. AB - We describe a 65-year-old Japanese man with a 20-year history of telangiectasia macularis eruptiva perstans, who developed polycythemia rubra vera and duodenal ulcer 10 and 12 years respectively after the onset of mastocytosis. Involvement of mast cells was found in neither bone marrow nor gastrointestinal tract. Immunohistochemical staining revealed that the mast cell was positive for both tryptase and chymase, indicating the nature of cutaneous mast cells. Despite the coexistence of a hematologic disorder, our case is suggested to have cutaneous but not systemic mastocytosis presenting as telangiectasia macularis eruptiva perstans. PMID- 11872426 TI - Guess what! Generalized eruptive histiocytosis (histiocytoma). PMID- 11872427 TI - Guess what! Multiple erythemato-hemorrhagic papules on the legs: papular dermatitis induced by Scleroderma domesticum. PMID- 11872428 TI - Guess what! Cellular variant of neurothekeoma. PMID- 11872432 TI - The evolving role of corporate medical departments in commercial aviation. AB - Dr. McKenas, who is Corporate Medical Director at American Airlines, describes some of AA s innovations in addressing on-board medical emergencies. These programs include Physician-on-Call and installation of automatic external defibrillators. The value of internal OM departments in matching medical services to changing traveler demographics, and in accident response such as was required on September 11, 2001, are also briefly discussed. PMID- 11872433 TI - Airline accident response. AB - This article outlines government regulations affecting accident response and offers guidelines for airline contingency plans in the face of major air disasters, such as those encountered on September 11, 2001. The author also touches upon the role of the corporate medical department in accident investigation and victim identification. PMID- 11872434 TI - The role of trauma debriefing in occupational medicine at an airline. AB - Most airlines now have formally established Critical Incident Management Programs. The critical incident stress debriefing (CISD) is a tool designed to relieve and prevent event-related stress. The author focuses on the use of CISD to assist victims of smaller traumas, such as assault, bomb threats, and severe turbulence, and shares the results of an American Airlines CIS program for flight attendants. PMID- 11872435 TI - Sudden incapacitation: occupational aviation medicine perspectives. AB - Within the aviation environment, occupational medicine tends to focus on pilots, because of the very great effects pilot injury or illness can have on public safety. However, medical conditions of other aviation workers can also endanger public and personal safety. Sudden incapacitation an abrupt loss or impairment of consciousness, control, or performance is the most important occupational concern in aviation medicine. The authors discuss the neurologic causes of sudden incapacitation and syncope, and evaluate the risks of returning affected pilots to duty. Also offered is a Controversy (Single Seizure: To Treat or Not To Treat). PMID- 11872436 TI - Toxicology and drug testing in aviation. AB - The occupational physician working in the aviation industry must have some toxicological expertise. Airline production and maintenance operations, while similar to other large manufacturing facilities, use some exotic metals and composites with unique toxicity. Airport operations involve exposure of the ground crew to de-icing chemicals and jet fuels. Moreover, evaluation of drug test results requires a background in pharmacology, physiology, and laboratory methods. Frank response to employees and coordination of plans with industrial hygienists, managers, and employees are necessary when toxicological questions arise. This article also offers a Controversy (Substituted Urine: Offer a Retest or Not?). PMID- 11872437 TI - The Federal Aviation Administration's role in evaluation of pilots and others with alcoholism or drug addiction. AB - The author explores the FAA Medical Standards, including old and new approaches toward rehabilitation; special issuance; and pilot initiatives. The current treatment model is also seen in various impaired physician programs around the U.S. PMID- 11872438 TI - Guidelines for the psychological evaluation of air crew personnel. AB - Air crew personnel (i.e., pilots, flight attendants, and air traffic controllers) are subjected to intense psychological and physical stress. Recent terrorist attacks in the U.S., the increased frequency of unruly passenger incidents, and the instability of the aviation industry have exacerbated air crew stress. In addition, common neuropsychiatric conditions (e.g., alcoholism, head trauma, HIV, depression) can impair operational fitness. This article discusses the aviation medical certification process and proposes guidelines for psychological evaluation, including: qualifications of providers, selection of psychological and neuropsychological tests, and the elements of a comprehensive, unbiased examination process. PMID- 11872439 TI - Alertness management in 24/7 settings: lessons from aviation. AB - Round-the-clock operational requirements pose physiological challenges for human operators. Fatigue due to sleep loss and circadian disruption can reduce safety, performance quality, and alertness. The authors describe the physiological factors underlying fatigue and provide examples from NASA research in aviation settings that demonstrate how fatigue affects real-world operations. A comprehensive alertness management approach to address fatigue effectively includes education, alertness strategies, scheduling, policy, and healthy sleep components. There is a need for cultural change that will encourage attitudes, behaviors, and practices that will reduce fatigue-related risks and improve safety, performance, and alertness in 24/7 operational settings. PMID- 11872440 TI - Medical standards for airmen and air traffic controllers. AB - This article details aeromedical certification requirements for first-, second-, and third-class airmen and reviews special issuance, including the administrative process regarding disease-specific special issuance. PMID- 11872441 TI - Airliner cabin air quality. AB - Airliner cabin air quality has attracted a lot of attention in the last two decades and has generated much controversy. This article reviews the situation to date only as it pertains to cabin air quality during normal aircraft operation. First discussed are the basic assumptions from the early days of commercial aviation. Second, different factors involved in cabin air quality are reviewed in some detail. Evidence from the current literature is employed to demonstrate that airliner cabin air quality is adequate and does not compromise aircrew health. However, the author does recognize that further studies are required for additional clarification of the issue. PMID- 11872442 TI - Radiation exposure of aircrews. AB - Information is provided about the radiation to which aircrews are exposed and possible health consequences. Recommended radiation exposure limits are given. Crewmembers on commercial aircraft are exposed to higher doses of ionizing radiation than normally received by members of the general population in most parts of the world. The principal ionizing radiation is galactic cosmic radiation. On infrequent occasions, radiation from the sun leads to an increase in the ionizing radiation at aircraft flight altitudes. Radioactive cargo is another possible source of exposure to ionizing radiation. Crewmembers are exposed to nonionizing radiation in the form of electric and magnetic fields generated by the aircraft s electronic and electrical systems. Other potential sources of nonionizing radiation exposure are microwave radiation from the aircraft's weather radar, laser radiation, and ultraviolet radiation. PMID- 11872443 TI - Aviation operations in foreign countries. AB - The authors make recommendations on personal medications and dental care, vaccinations, sanitation, and health coverage for airline personnel working outside the U.S. Also addressed are environmental demands and local customs. PMID- 11872444 TI - Handling an airline disaster: suggestions from psychological and logistical standpoints. AB - In light of the terrorist attacks on September 11, 2001, which eliminated all crew and passengers of four airliners and produced mass casualties on the ground, OM professionals must re-examine their crisis management programs to ensure that they are in the best possible shape. This article discusses the logistical components of such programs and stresses training, psychological debriefing and defusing, and reintegration of First Responders. PMID- 11872445 TI - Nitrogen cycling in the ocean: new perspectives on processes and paradigms. PMID- 11872446 TI - Chlorine disinfection of atypical mycobacteria isolated from a water distribution system. AB - We studied the resistance of various mycobacteria isolated from a water distribution system to chlorine. Chlorine disinfection efficiency is expressed as the coefficient of lethality (liters per minute per milligram) as follows: Mycobacterium fortuitum (0.02) > M. chelonae (0.03) > M. gordonae (0.09) > M. aurum (0.19). For a C.t value (product of the disinfectant concentration and contact time) of 60 mg.min.liter(-1), frequently used in water treatment lines, chlorine disinfection inactivates over 4 log units of M. gordonae and 1.5 log units of M. fortuitum or M. chelonae. C.t values determined under similar conditions show that even the most susceptible species, M. aurum and M. gordonae, are 100 and 330 times more resistant to chlorine than Escherichia coli. We also investigated the effects of different parameters (medium, pH, and temperature) on chlorine disinfection in a chlorine-resistant M. gordonae model. Our experimental results follow the Arrhenius equation, allowing the inactivation rate to be predicted at different temperatures. Our results show that M. gordonae is more resistant to chlorine in low-nutrient media, such as those encountered in water, and that an increase in temperature (from 4 degrees C to 25 degrees C) and a decrease in pH result in better inactivation. PMID- 11872447 TI - Development of a virus concentration method and its application to detection of enterovirus and norwalk virus from coastal seawater. AB - We developed a new procedure for concentration of enteric viruses from water using a negatively charged membrane. Rinsing the membrane with 0.5 mM H(2)SO(4) (pH 3.0) in order to elute cations prior to viral elution with 1 mM NaOH (pH 10.5) promoted poliovirus recovery yields from 33 to 95% when applied to pure water and 38 to 89% when applied to natural seawater from Tokyo Bay, Japan, respectively. This method showed average recovery yields of spiked poliovirus of 62% (n = 8) from 1 liter of artificial seawater. This method showed higher recovery yields (>61%) than that of the conventional method using positively charged membrane (6%) when applied to seawater. This method is also free from beef extract elution, which has an inhibitory effect in the subsequent viral genome detection by reverse transcription-PCR. Naturally occurring Norwalk viruses from 2 liters of Tokyo Bay water in winter and infectious enteroviruses from 2 liters of recreational coastal seawater in summer were detected by using this viral concentration method. PMID- 11872448 TI - Bacteriocin-like activity of Butyrivibrio fibrisolvens JL5 and its effect on other ruminal bacteria and ammonia production. AB - When ruminal bacteria from a cow fed hay were serially diluted into an anaerobic medium that had only peptides and amino acids as energy sources, little growth or ammonia production was detected at dilutions greater than 10(-6). The 10(-8) and 10(-9) dilutions contained bacteria that fermented carbohydrates, and some of these bacteria inhibited Clostridium sticklandii SR, an obligate amino acid fermenting bacterium. Phylogenetic analysis indicated that the most active isolate (JL5) was closely related to Butyrivibrio fibrisolvens B835. Strain JL5 inhibited B. fibrisolvens 49 and a variety of other gram-positive organisms, but it had little effect on most gram-negative ruminal bacteria. Strain JL5 did not produce a bacteriocin-like inhibitory substance (BLIS) until it reached the late log or stationary phase. The JL5 BLIS did not cause the lysis of B. fibrisolvens 49, but the intracellular potassium level, the ATP level, the electrical potential, and the viability decreased rapidly. The JL5 BLIS also caused marked decreases in the viability and cellular potassium level of C. sticklandii SR. The membrane potential and intracellular ATP level also declined. The BLIS was degraded very slowly by pronase E, but it could be precipitated with 60% ammonium sulfate and dialyzed (3,500-Da cutoff). The BLIS could be separated from other peptides by polyacrylamide gel electrophoresis, and C. sticklandii SR overlays indicated that the molecular size of this compound was approximately 3,600 Da. Based on these results, it appeared that the JL5 BLIS was a pore-forming peptide. Because carbohydrate-fermenting ruminal bacteria could inhibit the growth of obligate amino acid-fermenting bacteria, BLIS may play a role in regulating ammonia production in vivo. PMID- 11872450 TI - Two-dimensional electrophoresis study of Lactobacillus delbrueckii subsp. bulgaricus thermotolerance. AB - The response of Lactobacillus delbrueckii subsp. bulgaricus cells to heat stress was studied by use of a chemically defined medium. Two-dimensional electrophoresis (2-DE) analysis was used to correlate the kinetics of heat shock protein (HSP) induction with cell recovery from heat injury. We demonstrated that enhanced viability, observed after 10 min at 65 degrees C, resulted from the overexpression of HSP and from mechanisms not linked to protein synthesis. In order to analyze the thermoadaptation mechanisms involved, thermoresistant variants were selected. These variants showed enhanced constitutive tolerance toward heat shock. However, contrary to the wild-type strain, these variants were poorly protected after osmotic or heat pretreatments. This result suggests that above a certain threshold, cells reach a maximum level of protection that cannot be easily exceeded. A comparison of protein patterns showed that the variants were able to induce more rapidly their adaptive mechanisms than the original strain. In particular, the variants were able to express constitutively more HSP, leading to the higher level of thermoprotection observed. This is the first report of the study by 2-DE of the heat stress response in L. delbrueckii subsp. bulgaricus. PMID- 11872449 TI - Characterization of a chitinase gene from Stenotrophomonas maltophilia strain 34S1 and its involvement in biological control. AB - A chitinase gene was cloned on a 2.8-kb DNA fragment from Stenotrophomonas maltophilia strain 34S1 by heterologous expression in Burkholderia cepacia. Sequence analysis of this fragment identified an open reading frame encoding a deduced protein of 700 amino acids. Removal of the signal peptide sequence resulted in a predicted protein that was 68 kDa in size. Analysis of the sequence indicated that the chitinase contained a catalytic domain belonging to family 18 of glycosyl hydrolases. Three putative binding domains, a chitin binding domain, a novel polycystic kidney disease (PKD) domain, and a fibronectin type III domain, were also identified within the sequence. Pairwise comparisons of each domain to the most closely related sequences found in database searches clearly demonstrated variation in gene sources and the species from which related sequences originated. A 51-kDa protein with chitinolytic activity was purified from culture filtrates of S. maltophilia strain 34S1 by hydrophobic interaction chromatography. Although the protein was significantly smaller than the size predicted from the sequence, the N-terminal sequence verified that the first 15 amino acids were identical to the deduced sequence of the mature protein encoded by chiA. Marker exchange mutagenesis of chiA resulted in mutant strain C5, which was devoid of chitinolytic activity and lacked the 51-kDa protein in culture filtrates. Strain C5 was also reduced in the ability to suppress summer patch disease on Kentucky bluegrass, supporting a role for the enzyme in the biocontrol activity of S. maltophilia. PMID- 11872451 TI - Morphological changes of rhizobia in peat cultures. AB - Morphological changes that take place in peat cultures of several species of rhizobia were examined. These changes seemed to be associated with enhanced survival of cells in peat and after inoculation onto plastic beads, which were used as a model system for seeds. Cell wall changes, in which the periplasmic space appeared to be occluded with electron-dense material, were observed in Rhizobium sp. strain SU343 and Bradyrhizobium lupini WU425 cells after 7 and 14 days in peat, respectively. Nutrient limitation and low O(2) concentration in peat are suggested to be factors involved in the induction of the morphological changes. Polyhydroxybutyrate reserves, which were present in broth-cultured cells of both species of rhizobia, were mobilized after transfer into peat but did not appear to influence survival after inoculation onto beads. Enhanced expression of an iron-manganese superoxide dismutase was also observed after the cells were transferred into peat. We conclude that cell wall thickening in rhizobia after transfer from broth cultures into peat is an adaptive response for long-term survival under nutrient-limited conditions in peat. Cells with thickened walls may also be more resistant to other types of stress, such as that encountered on a seed surface. PMID- 11872453 TI - Simultaneous extraction from bacterioplankton of total RNA and DNA suitable for quantitative structure and function analyses. AB - The aim of this study was to develop a protocol for the simultaneous extraction from bacterioplankton of RNA and DNA suitable for quantitative molecular analysis. By using a combined mechanical and chemical extraction method, the highest RNA and DNA yield was obtained with sodium lauryl sarcosinate-phenol or DivoLab-phenol as the extraction mix. The efficiency of extraction of nucleic acids was comparatively high and varied only moderately in gram-negative bacterial isolates and bacterioplankton (RNA, 52 to 66%; DNA, 43 to 61%); significant amounts of nucleic acids were also obtained for a gram-positive bacterial isolate (RNA, 20 to 30%; DNA, 20 to 25%). Reverse transcription-PCR and PCR amplification products of fragments of 16S rRNA and its genes were obtained from all isolates and communities, indicating that the extracted nucleic acids were intact and pure enough for community structure analyses. By using single strand conformation polymorphism of fragments of 16S rRNA and its gene, community fingerprints were obtained from pond bacterioplankton. mRNA transcripts encoding fragments of the enzyme nitrite reductase gene (nir gene) could be detected in a pond water sample, indicating that the extraction method is also suitable for studying gene expression. The extraction method presented yields nucleic acids that can be used to perform structural and functional studies of bacterioplankton communities from a single sample. PMID- 11872452 TI - Flux through citrate synthase limits the growth of ethanologenic Escherichia coli KO11 during xylose fermentation. AB - Previous studies have shown that high levels of complex nutrients (Luria broth or 5% corn steep liquor) were necessary for rapid ethanol production by the ethanologenic strain Escherichia coli KO11. Although this strain is prototrophic, cell density and ethanol production remained low in mineral salts media (10% xylose) unless complex nutrients were added. The basis for this nutrient requirement was identified as a regulatory problem created by metabolic engineering of an ethanol pathway. Cells must partition pyruvate between competing needs for biosynthesis and regeneration of NAD(+). Expression of low K(m) Zymomonas mobilis pdc (pyruvate decarboxylase) in KO11 reduced the flow of pyruvate carbon into native fermentation pathways as desired, but it also restricted the flow of carbon skeletons into the 2-ketoglutarate arm of the tricarboxylic acid pathway (biosynthesis). In mineral salts medium containing 1% corn steep liquor and 10% xylose, the detrimental effect of metabolic engineering was substantially reduced by addition of pyruvate. A similar benefit was also observed when acetaldehyde, 2-ketoglutarate, or glutamate was added. In E. coli, citrate synthase links the cellular abundance of NADH to the supply of 2 ketoglutarate for glutamate biosynthesis. This enzyme is allosterically regulated and inhibited by high NADH concentrations. In addition, citrate synthase catalyzes the first committed step in 2-ketoglutarate synthesis. Oxidation of NADH by added acetaldehyde (or pyruvate) would be expected to increase the activity of E. coli citrate synthase and direct more carbon into 2-ketoglutarate, and this may explain the stimulation of growth. This hypothesis was tested, in part, by cloning the Bacillus subtilis citZ gene encoding an NADH-insensitive citrate synthase. Expression of recombinant citZ in KO11 was accompanied by increases in cell growth and ethanol production, which substantially reduced the need for complex nutrients. PMID- 11872454 TI - Effects of pressure-induced membrane phase transitions on inactivation of HorA, an ATP-dependent multidrug resistance transporter, in Lactobacillus plantarum. AB - The effects of pressure on cultures of Lactobacillus plantarum were characterized by determination of the viability and activity of HorA, an ATP-binding cassette multidrug resistance transporter. Changes in the membrane composition of L. plantarum induced by different growth temperatures were determined. Furthermore, the effect of the growth temperature of a culture on pressure inactivation at 200 MPa was determined. Cells were characterized by plate counts on selective and nonselective agar after pressure treatment, and HorA activity was measured by ethidium bromide efflux. Fourier transform-infrared spectroscopy and Laurdan fluorescence spectroscopy provided information about the thermodynamic phase state of the cytoplasmic membrane during pressure treatment. A pressure temperature diagram for cell membranes was established. Cells grown at 37 degrees C and pressure treated at 15 degrees C lost >99% of HorA activity and viable cell counts within 36 and 120 min, respectively. The membranes of these cells were in the gel phase region at ambient pressure. In contrast, cells grown at 15 degrees C and pressure treated at 37 degrees C lost >99% of HorA activity and viable cell counts within 4 and 8 min, respectively. The membranes of these cells were in the liquid crystalline phase region at ambient pressure. The kinetic analysis of inactivation of L. plantarum provided further evidence that inactivation of HorA is a crucial step during pressure-induced cell death. Comparison of the biological findings and the membrane state during pressure treatment led to the conclusion that the inactivation of cells and membrane enzymes strongly depends on the thermodynamic properties of the membrane. Pressure treatment of cells with a liquid crystalline membrane at 0.1 MPa resulted in HorA inactivation and cell death more rapid than those of cells with a gel phase membrane at 0.1 MPa. PMID- 11872455 TI - Population dynamics of active and total ciliate populations in arable soil amended with wheat. AB - Soil protozoa are characterized by their ability to produce cysts, which allows them to survive unfavorable conditions (e.g., desiccation) for extended periods. Under favorable conditions, they may rapidly excyst and begin feeding, but even under optimal conditions, a large proportion of the population may be encysted. The factors governing the dynamics of active and encysted cells in the soil are not well understood. Our objective was to determine the dynamics of active and encysted populations of ciliates during the decomposition of freshly added organic material. We monitored, in soil microcosms, the active and total populations of ciliates, their potential prey (bacteria and small protozoa), their potential competitors (amoebae, flagellates, and nematodes), and their potential predators (nematodes). We sampled with short time intervals (2 to 6 days) and generated a data set, suitable for mathematical modeling. Following the addition of fresh organic material, bacterial numbers increased more than 1,400 fold. There was a temporary increase in the number of active ciliates, followed by a rapid decline, although the size of the bacterial prey populations remained high. During this initial burst of ciliate growth, the population of cystic ciliates increased 100-fold. We suggest that internal population regulation is the major factor governing ciliate encystment and that the rate of encystment depends on ciliate density. This model provides a quantitative explanation of ciliatostasis and can explain why protozoan growth in soil is less than that in aquatic systems. Internally governed encystment may be an essential adaptation to an unpredictable environment in which individual protozoa cannot predict when the soil will dry out and will survive desiccation only if they have encysted in time. PMID- 11872456 TI - Engineering of a Bacillus subtilis strain with adjustable levels of intracellular biotin for secretory production of functional streptavidin. AB - Streptavidin is a biotin-binding protein which has been widely used in many in vitro and in vivo applications. Because of the ease of protein recovery and availability of protease-deficient strains, the Bacillus subtilis expression secretion system is an attractive system for streptavidin production. However, attempts to produce streptavidin using B. subtilis face the problem that cells overproducing large amounts of streptavidin suffer poor growth, presumably because of biotin deficiency. This problem cannot be solved by supplementing biotin to the culture medium, as this will saturate the biotin binding sites in streptavidin. We addressed this dilemma by engineering a B. subtilis strain (WB800BIO) which overproduces intracellular biotin. The strategy involves replacing the natural regulatory region of the B. subtilis chromosomal biotin biosynthetic operon (bioWAFDBIorf2) with an engineered one consisting of the B. subtilis groE promoter and gluconate operator. Biotin production in WB800BIO is induced by gluconate, and the level of biotin produced can be adjusted by varying the gluconate dosage. A level of gluconate was selected to allow enhanced intracellular production of biotin without getting it released into the culture medium. WB800BIO, when used as a host for streptavidin production, grows healthily in a biotin-limited medium and produces large amounts (35 to 50 mg/liter) of streptavidin, with over 80% of its biotin binding sites available for future applications. PMID- 11872457 TI - Quantitative trait loci controlling vegetative growth rate in the edible basidiomycete Pleurotus ostreatus. AB - Mycelium growth rate is a quantitative characteristic that exhibits continuous variation. This trait has applied interest, as growth rate is correlated with production yield and increased advantage against competitors. In this work, we studied growth rate variation in the edible basidiomycete Pleurotus ostreatus growing as monokaryotic or dikaryotic mycelium on Eger medium or on wheat straw. Our analysis resulted in identification of several genomic regions (quantitative trait loci [QTLs]) involved in the control of growth rate that can be mapped on the genetic linkage map of this fungus. In some cases monokaryotic and dikaryotic QTLs clustered at the same map position, indicating that there are principal genomic areas responsible for growth rate control. The availability of this linkage map of growth rate QTLs can help in the design of rational strain breeding programs based on genomic information. PMID- 11872458 TI - Identification of Cryptosporidium parvum oocysts by an artificial neural network approach. AB - Microscopic detection of Cryptosporidium parvum oocysts is time-consuming, requires trained analysts, and is frequently subject to significant human errors. Artificial neural networks (ANN) were developed to help identify immunofluorescently labeled C. parvum oocysts. A total of 525 digitized images of immunofluorescently labeled oocysts, fluorescent microspheres, and other miscellaneous nonoocyst images were employed in the training of the ANN. The images were cropped to a 36- by 36-pixel image, and the cropped images were placed into two categories, oocyst and nonoocyst images. The images were converted to grayscale and processed into a histogram of gray color pixel intensity. Commercially available software was used to develop and train the ANN. The networks were optimized by varying the number of training images, number of hidden neurons, and a combination of these two parameters. The network performance was then evaluated using a set of 362 unique testing images which the network had never "seen" before. Under optimized conditions, the correct identification of authentic oocyst images ranged from 81 to 97%, and the correct identification of nonoocyst images ranged from 78 to 82%, depending on the type of fluorescent antibody that was employed. The results indicate that the ANN developed were able to generalize the training images and subsequently discern previously unseen oocyst images efficiently and reproducibly. Thus, ANN can be used to reduce human errors associated with the microscopic detection of Cryptosporidium oocysts. PMID- 11872459 TI - Sunlight inactivation of fecal indicator bacteria and bacteriophages from waste stabilization pond effluent in fresh and saline waters. AB - Sunlight inactivation in fresh (river) water of fecal coliforms, enterococci, Escherichia coli, somatic coliphages, and F-RNA phages from waste stabilization pond (WSP) effluent was compared. Ten experiments were conducted outdoors in 300 liter chambers, held at 14C (mean river water temperature). Sunlight inactivation (k(S)) rates, as a function of cumulative global solar radiation (insolation), were all more than 10 times higher than the corresponding dark inactivation (k(D)) rates in enclosed (control) chambers. The overall k(S) ranking (from greatest to least inactivation) was as follows: enterococci > fecal coliforms greater-than-or-equal E. coli > somatic coliphages > F-RNA phages. In winter, fecal coliform and enterococci inactivation rates were similar but, in summer, enterococci were inactivated far more rapidly. In four experiments that included freshwater-raw sewage mixtures, enterococci survived longer than fecal coliforms (a pattern opposite to that observed with the WSP effluent), but there was little difference in phage inactivation between effluents. In two experiments which included simulated estuarine water and seawater, sunlight inactivation of all of the indicators increased with increasing salinity. Inactivation rates in freshwater, as seen under different optical filters, decreased with the increase in the spectral cutoff (50% light transmission) wavelength. The enterococci and F RNA phages were inactivated by a wide range of wavelengths, suggesting photooxidative damage. Inactivation of fecal coliforms and somatic coliphages was mainly by shorter (UV-B) wavelengths, a result consistent with photobiological damage. Fecal coliform repair mechanisms appear to be activated in WSPs, and the surviving cells exhibit greater sunlight resistance in natural waters than those from raw sewage. In contrast, enterococci appear to suffer photooxidative damage in WSPs, rendering them susceptible to further photooxidative damage after discharge. This suggests that they are unsuitable as indicators of WSP effluent discharges to natural waters. Although somatic coliphages are more sunlight resistant than the other indicators in seawater, F-RNA phages are the most resistant in freshwater, where they may thus better represent enteric virus survival. PMID- 11872460 TI - The aquatic budding bacterium Blastobacter denitrificans is a nitrogen-fixing symbiont of Aeschynomene indica. AB - Blastobacter spp. are freshwater bacteria that form rosette structures by cellular attachment to a common base. Comparative analyses of ribosomal 16S rRNA gene and internally transcribed spacer region sequences indicated that B. denitrificans is a member of the alpha-subdivision of Proteobacteria. Among the alpha-Proteobacteria, B. denitrificans was related to a cluster of genera, including Rhodopseudomonas palustris, Afipia felis, Nitrobacter hamburgensis, and Bradyrhizobium spp. Although the precise phylogenetic relationships among these genera could not be established with a high degree of confidence, the sequences of B. denitrificans and several bradyrhizobial isolates from nodules of Aeschynomene indica were almost identical. Bradyrhizobia are bacteria that form nitrogen-fixing symbioses with legumes, including soybeans (Glycine max) and members of the genus Aeschynomene. From symbiotic infectiveness tests we demonstrated that the type strain for B. denitrificans, IFAM 1005, was capable of forming an effective nitrogen-fixing symbiosis with A. indica. Not only do these results reveal a previously unknown ecological adaptation of a relatively obscure aquatic bacterium, but they also demonstrate how evidence gathered from molecular systematic analyses can sometimes provide clues for predicting ecological behavior. PMID- 11872461 TI - Novel Bacillus thuringiensis binary insecticidal crystal proteins active on western corn rootworm, Diabrotica virgifera virgifera LeConte. AB - A new family of insecticidal crystal proteins was discovered by screening sporulated Bacillus thuringiensis cultures for oral activity against western corn rootworm (WCR) larvae. B. thuringiensis isolates PS80JJ1, PS149B1, and PS167H2 have WCR insecticidal activity attributable to parasporal inclusion bodies containing proteins with molecular masses of ca. 14 and 44 kDa. The genes encoding these polypeptides reside in apparent operons, and the 14-kDa protein open reading frame (ORF) precedes the 44-kDa protein ORF. Mutagenesis of either gene in the apparent operons dramatically reduced insecticidal activity of the corresponding recombinant B. thuringiensis strain. Bioassays performed with separately expressed, biochemically purified 14- and 44-kDa polypeptides also demonstrated that both proteins are required for WCR mortality. Sequence comparisons with other known B. thuringiensis insecticidal proteins failed to reveal homology with previously described Cry, Cyt, or Vip proteins. However, there is evidence that the 44-kDa polypeptide and the 41.9- and 51.4-kDa binary dipteran insecticidal proteins from Bacillus sphaericus are evolutionarily related. The 14- and 44-kDa polypeptides from isolates PS80JJ1, PS149B1, and PS167H2 have been designated Cry34Aa1, Cry34Ab1, and Cry34Ac1, respectively, and the 44-kDa polypeptides from these isolates have been designated Cry35Aa1, Cry35Ab1, and Cry35Ac1, respectively. PMID- 11872462 TI - Development and evaluation of a 16S ribosomal DNA array-based approach for describing complex microbial communities in ready-to-eat vegetable salads packed in a modified atmosphere. AB - There is a clear need for new approaches in the field of microbial community analyses, since the methods used can be severely biased. We have developed a DNA array-based method that targets 16S ribosomal DNA (rDNA), enabling the direct detection and quantification of microorganisms from complex communities without cultivation. The approach is based on the construction of specific probes from the 16S rDNA sequence data retrieved directly from the communities. The specificity of the assay is obtained through a combination of DNA array hybridization and enzymatic labeling of the constructed probes. Cultivation dependent assays (enrichment and plating) and cultivation-independent assays (direct fluorescence microscopy and scanning electron microscopy) were used as reference methods in the development and evaluation of the method. The description of microbial communities in ready-to-eat vegetable salads in a modified atmosphere was used as the experimental model. Comparisons were made with respect to the effect of storage at different temperatures for up to 12 days and with respect to the geographic origin of the crisphead lettuce (Spanish or Norwegian), the main salad component. The conclusion drawn from the method comparison was that the DNA array-based method gave an accurate description of the microbial communities. Pseudomonas spp. dominated both of the salad batches, containing either Norwegian or Spanish lettuce, before storage and after storage at 4 degrees C. The Pseudomonas population also dominated the batch containing Norwegian lettuce after storage at 10 degrees C. On the contrary, Enterobacteriaceae and lactic acid bacteria dominated the microbial community of the batch containing Spanish lettuce after storage at 10 degrees C. In that batch, the Enterobacteriaceae also were abundant after storage at 4 degrees C as well as before storage. The practical implications of these results are that microbial communities in ready-to-eat vegetable salads can be diverse and that microbial composition is dependent both on the origin of the raw material and on the storage conditions. PMID- 11872463 TI - Interactions among plant species and microorganisms in salt marsh sediments. AB - The interactions among Spartina patens and sediment microbial populations and the interactions among Phragmites australis and sediment microbial populations were studied at monotypic sites in Piermont Marsh, a salt marsh of the Hudson River north of New York, N.Y., at key times during the growing season. Arbuscular mycorrhizal fungi (AMF) effectively colonized S. patens but not P. australis, and there were seasonal increases and decreases that coincided with plant growth and senescence (17 and 6% of the S. patens root length were colonized, respectively). In sediment samples from the Spartina site, the microbial community and specific bacterial populations were at least twice as large in terms of number and biomass as the microbial community and specific bacterial populations in sediment samples from the Phragmites site, and peak values occurred during reproduction. Members of the domain Bacteria, especially members of the alpha-, gamma-, and delta subdivisions of the Proteobacteria, were the most abundant organisms at both sites throughout the growing season. The populations were generally more dynamic in samples from the Spartina site than in samples from the Phragmites site. No differences between the two sites and no differences during the growing season were observed when restriction fragment length polymorphism analyses of nifH amplicons were performed in an attempt to detect shifts in the diversity of nitrogen-fixing bacteria. Differences were observed only in the patterns generated by PCR or reverse transcription-PCR for samples from the Spartina site, suggesting that there were differences in the overall and active populations of nitrogen-fixing bacteria. Regression analyses indicated that there was a positive interaction between members of the delta-subdivision of the Proteobacteria and root biomass but not between members of the delta-subdivision of the Proteobacteria and macroorganic matter at both sites. In samples from the Spartina site, there were indications that there were bacterium-fungus interactions since populations of members of the alpha-subdivision of the Proteobacteria were negatively associated with AMF colonization and populations of members of the gamma-subdivision of the Proteobacteria were positively associated with AMF colonization. PMID- 11872464 TI - Influence of soil on fecal indicator organisms in a tidally influenced subtropical environment. AB - The potential regrowth of fecal indicator bacteria released into coastal environments in recreational water bodies has been of concern, especially in tropical and subtropical areas where the number of these bacteria can be artificially elevated beyond that from fecal impacts alone. The task of determining the factors that influence indicator bacterial regrowth was addressed though a series of field sampling and laboratory experiments using in situ densities of Escherichia coli, enterococci, and Clostridium perfringens in river water, sediment, and soil. Field sampling efforts included the collection of surface sediments along the cross section of a riverbank, a 20-cm-deep soil core, and additional surface soils from remote locations. In addition to field sampling, two types of laboratory experiments were conducted. The first experiment investigated the survival of bacteria already present in river water with the addition of sterile and unsterile sediment. The second experiment was designed to simulate the wetting and drying effects due to tidal cycles. The results from the sampling study found elevated numbers of E. coli and C. perfringens in surficial sediments along the riverbank near the edge of the water. C. perfringens was found in high numbers in the subsurface samples obtained from the soil core. Results from laboratory experiments revealed a significant amount of regrowth for enterococci and E. coli with the simulation of tides and addition of sterile sediment. Regrowth was not observed for C. perfringens. This study demonstrates the need to further evaluate the characteristics of indicator microbes within tropical and subtropical water systems where natural vegetation, soil embankments, and long-term sediment accumulation are present. In such areas, the use of traditional indicator microbes to regulate recreational uses of a water body may not be appropriate. PMID- 11872465 TI - Differential expression of methanogenesis genes of Methanothermobacter thermoautotrophicus (formerly Methanobacterium thermoautotrophicum) in pure culture and in cocultures with fatty acid-oxidizing syntrophs. AB - The expression of genes involved in methanogenesis in a thermophilic hydrogen utilizing methanogen, Methanothermobacter thermoautotrophicus strain TM, was investigated both in a pure culture sufficiently supplied with H(2) plus CO(2) and in a coculture with an acetate-oxidizing hydrogen-producing bacterium, Thermacetogenium phaeum strain PB, in which hydrogen partial pressure was constantly kept very low (20 to 80 Pa). Northern blot analysis indicated that only the mcr gene, which encodes methyl coenzyme M reductase I (MRI), catalyzing the final step of methanogenesis, was expressed in the coculture, whereas mcr and mrt, which encodes methyl coenzyme M reductase II (MRII), the isofunctional enzyme of MRI, were expressed at the early to late stage of growth in the pure culture. In contrast to these two genes, two isofunctional genes (mtd and mth) for N(5),N(10)-methylene-tetrahydromethanopterin dehydrogenase, which catalyzes the fourth step of methanogenesis, and two hydrogenase genes (frh and mvh) were expressed both in a pure culture and in a coculture at the early and late stages of growth. The same expression pattern was observed for Methanothermobacter thermoautotrophicus strain DeltaH cocultured with a thermophilic butyrate oxidizing syntroph, Syntrophothermus lipocalidus strain TGB-C1. Two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis of whole proteins of M. thermoautotrophicus strain TM obtained from a pure culture and a coculture with the acetate-oxidizing syntroph and subsequent N-terminal amino acid sequence analysis confirmed that MRI and MRII were produced in the pure culture, while only MRI was produced in the coculture. These results indicate that under syntrophic growth conditions, the methanogen preferentially utilizes MRI but not MRII. Considering that hydrogenotrophic methanogens are strictly dependent for growth on hydrogen-producing fermentative microbes in the natural environment and that the hydrogen supply occurs constantly at very low concentrations compared with the supply in pure cultures in the laboratory, the results suggest that MRI is an enzyme primarily functioning in natural methanogenic ecosystems. PMID- 11872466 TI - Resolution of Prochlorococcus and Synechococcus ecotypes by using 16S-23S ribosomal DNA internal transcribed spacer sequences. AB - Cultured isolates of the marine cyanobacteria Prochlorococcus and Synechococcus vary widely in their pigment compositions and growth responses to light and nutrients, yet show greater than 96% identity in their 16S ribosomal DNA (rDNA) sequences. In order to better define the genetic variation that accompanies their physiological diversity, sequences for the 16S-23S rDNA internal transcribed spacer (ITS) region were determined in 32 Prochlorococcus isolates and 25 Synechococcus isolates from around the globe. Each strain examined yielded one ITS sequence that contained two tRNA genes. Dramatic variations in the length and G+C content of the spacer were observed among the strains, particularly among Prochlorococcus strains. Secondary-structure models of the ITS were predicted in order to facilitate alignment of the sequences for phylogenetic analyses. The previously observed division of Prochlorococcus into two ecotypes (called high and low-B/A after their differences in chlorophyll content) were supported, as was the subdivision of the high-B/A ecotype into four genetically distinct clades. ITS-based phylogenies partitioned marine cluster A Synechococcus into six clades, three of which can be associated with a particular phenotype (motility, chromatic adaptation, and lack of phycourobilin). The pattern of sequence divergence within and between clades is suggestive of a mode of evolution driven by adaptive sweeps and implies that each clade represents an ecologically distinct population. Furthermore, many of the clades consist of strains isolated from disparate regions of the world's oceans, implying that they are geographically widely distributed. These results provide further evidence that natural populations of Prochlorococcus and Synechococcus consist of multiple coexisting ecotypes, genetically closely related but physiologically distinct, which may vary in relative abundance with changing environmental conditions. PMID- 11872467 TI - Wax ester production from n-alkanes by Acinetobacter sp. strain M-1: ultrastructure of cellular inclusions and role of acyl coenzyme A reductase. AB - Acinetobacter sp. strain M-1 accumulated a large amount of wax esters from an n alkane under nitrogen-limiting conditions. Under the optimized conditions with n hexadecane as the substrate, the amount of hexadecyl hexadecanoate in the cells reached 0.17 g/g of cells (dry weight). Electron microscopic analysis revealed that multilayered disk-shaped intracellular inclusions were formed concomitant with wax ester formation. The contribution of acyl-CoA reductase to wax ester synthesis was evaluated by gene disruption analysis. PMID- 11872469 TI - Microscale biosensor for measurement of volatile fatty acids in anoxic environments. AB - A microscale biosensor for acetate, propionate, isobutyrate, and lactate is described. The sensor is based on the bacterial respiration of low-molecular weight, negatively charged species with a concomitant reduction of NO(-)(3) to N(2)O. A culture of denitrifying bacteria deficient in N(2)O reductase was immobilized in front of the tip of an electrochemical N(2)O microsensor. The bacteria were separated from the outside environment by an ion-permeable membrane and supplied with nutrients (except for electron donors) from a medium reservoir behind the N(2)O sensor. The signal of the sensor, which corresponded to the rate of N(2)O production, was proportional to the supply of the electron donor to the bacterial mass. The selectivity for volatile fatty acids compared to other organic compounds was increased by selectively enhancing the transport of negatively charged compounds into the sensor by electrophoretic migration (electrophoretic sensitivity control). The sensor was susceptible to interference from O(2), N(2)O, NO(2)(-), H(2)S, and NO(-)(3). Interference from NO(-)(3) was low and could be quantified and accounted for. The detection limit was equivalent to about 1 microM acetate, and the 90% response time was 30 to 90 s. The response of the sensor was not affected by changes in pH between 5.5 and 9 and was also unaffected by changes in salinity in the range of 2 to 32 per thousand. The functioning of the sensor over a temperature span of 7 to 30 degrees C was investigated. The concentration range for a linear response was increased five times by increasing the temperature from 7 to 19.5 degrees C. The life span of the biosensor varied between 1 and 3 weeks after manufacturing. PMID- 11872468 TI - Functional genomics approach to identifying genes required for biofilm development by Streptococcus mutans. AB - Streptococcus mutans, the primary etiological agent of human dental caries, is an obligate biofilm-forming bacterium. The goals of this study were to identify the gene(s) required for biofilm formation by this organism and to elucidate the role(s) that some of the known global regulators of gene expression play in controlling biofilm formation. In S. mutans UA159, the brpA gene (for biofilm regulatory protein) was found to encode a novel protein of 406 amino acid residues. A strain carrying an insertionally inactivated copy of brpA formed longer chains than did the parental strain, aggregated in liquid culture, and was unable to form biofilms as shown by an in vitro biofilm assay. A putative homologue of the enzyme responsible for synthesis of autoinducer II (AI-2) of the bacterial quorum-sensing system was also identified in S. mutans UA159, but insertional inactivation of the gene (luxS(Sm)) did not alter colony or cell morphology or diminish the capacity of S. mutans to form biofilms. We also examined the role of the homologue of the Bacillus subtilis catabolite control protein CcpA in S. mutans in biofilm formation, and the results showed that loss of CcpA resulted in about a 60% decrease in the ability to form biofilms on an abiotic surface. From these data, we conclude that CcpA and BrpA may regulate genes that are required for stable biofilm formation by S. mutans. PMID- 11872471 TI - Nucleotide sequence and genetic structure of a novel carbaryl hydrolase gene (cehA) from Rhizobium sp. strain AC100. AB - Rhizobium sp. strain AC100, which is capable of degrading carbaryl (1-naphthyl-N methylcarbamate), was isolated from soil treated with carbaryl. This bacterium hydrolyzed carbaryl to 1-naphthol and methylamine. Carbaryl hydrolase from the strain was purified to homogeneity, and its N-terminal sequence, molecular mass (82 kDa), and enzymatic properties were determined. The purified enzyme hydrolyzed 1-naphthyl acetate and 4-nitrophenyl acetate indicating that the enzyme is an esterase. We then cloned the carbaryl hydrolase gene (cehA) from the plasmid DNA of the strain and determined the nucleotide sequence of the 10-kb region containing cehA. No homologous sequences were found by a database homology search using the nucleotide and deduced amino acid sequences of the cehA gene. Six open reading frames including the cehA gene were found in the 10-kb region, and sequencing analysis shows that the cehA gene is flanked by two copies of insertion sequence-like sequence, suggesting that it makes part of a composite transposon. PMID- 11872470 TI - Production of penicillin by fungi growing on food products: identification of a complete penicillin gene cluster in Penicillium griseofulvum and a truncated cluster in Penicillium verrucosum. AB - Mycobiota growing on food is often beneficial for the ripening and development of the specific flavor characteristics of the product, but it can also be harmful due to the production of undesirable compounds such as mycotoxins or antibiotics. Some of the fungi most frequently isolated from fermented and cured meat products such as Penicillium chrysogenum and Penicillium nalgiovense are known penicillin producers; the latter has been shown to be able to produce penicillin when growing on the surface of meat products and secrete it to the medium. The presence of penicillin in food must be avoided, since it can lead to allergic reactions and the arising of penicillin resistance in human-pathogenic bacteria. In this article we describe a study of the penicillin production ability among fungi of the genus Penicillium that are used as starters for cheese and meat products or that are frequently isolated from food products. Penicillium griseofulvum was found to be a new penicillin producer and to have a penicillin gene cluster similar to that of Penicillium chrysogenum. No other species among the studied fungi were found to produce penicillin or to possess the penicillin biosynthetic genes, except P. verrucosum, which contains the pcbAB gene (as shown by hybridization and PCR cloning of fragments of the gene) but lacks pcbC and penDE. Antibacterial activities due to the production of secondary metabolites other than penicillin were observed in some fungi. PMID- 11872472 TI - Characterization of Cyt2Bc toxin from Bacillus thuringiensis subsp. medellin. AB - We cloned and sequenced a new cytolysin gene from Bacillus thuringiensis subsp. medellin. Three IS240-like insertion sequence elements and the previously cloned cyt1Ab and p21 genes were found in the vicinity of the cytolysin gene. The cytolysin gene encodes a protein 29.7 kDa in size that is 91.5% identical to Cyt2Ba from Bacillus thuringiensis subsp. israelensis and has been designated Cyt2Bc. Inclusions containing Cyt2Bc were purified from the crystal-negative strain SPL407 of B. thuringiensis. Cyt2Bc reacted weakly with antibodies directed against Cyt2Ba and was not recognized by an antiserum directed against the reference cytolysin Cyt1Aa. Cyt2Bc was hemolytic only upon activation with trypsin and had only one-third to one-fifth of the activity of Cyt2Ba, depending on the activation time. Cyt2Bc was also mosquitocidal against Aedes aegypti, Anopheles stephensi, and Culex quinquefasciatus, including strains resistant to the Bacillus sphaericus binary toxin. Its toxicity was half of that of Cyt2Ba on all mosquito species except resistant C. quinquefasciatus. PMID- 11872473 TI - Molecular cloning of XYL3 (D-xylulokinase) from Pichia stipitis and characterization of its physiological function. AB - XYL3, which encodes a D-xylulokinase (EC 2.7.1.17), was isolated from Pichia stipitis CBS 6054 genomic DNA by using primers designed against conserved motifs. Disruption of XYL3 eliminated D-xylulokinase activity, but D-ribulokinase activity was still present. Southern analysis of P. stipitis genomic DNA with XYL3 as a probe confirmed the disruption and did not reveal additional related genes. Disruption of XYL3 stopped ethanol production from xylose, but the resulting mutant still assimilated xylose slowly and formed xylitol and arabinitol. These results indicate that XYL3 is critical for ethanol production from xylose but that P. stipitis has another pathway for xylose assimilation. Expression of XYL3 using its P. stipitis promoter increased Saccharomyces cerevisiae D-xylulose consumption threefold and enabled the transformants to produce ethanol from a mixture of xylose and xylulose, whereas the parental strain only accumulated xylitol. In vitro, D-xylulokinase activity in recombinant S. cerevisiae was sixfold higher with a multicopy than with a single-copy XYL3 plasmid, but ethanol production decreased with increased copy number. These results confirmed the function of XYL3 in S. cerevisiae. PMID- 11872474 TI - Nitrous oxide formation in the Colne estuary, England: the central role of nitrite. AB - Nitrate and nitrite concentrations in the water and nitrous oxide and nitrite fluxes across the sediment-water interface were measured monthly in the River Colne estuary, England, from December 1996 to March 1998. Water column concentrations of N(2)O in the Colne were supersaturated with respect to air, indicating that the estuary was a source of N(2)O for the atmosphere. At the freshwater end of the estuary, nitrous oxide effluxes from the sediment were closely correlated with the nitrite concentrations in the overlying water and with the nitrite influx into the sediment. Increases in N(2)O production from sediments were about 10 times greater with the addition of nitrite than with the addition of nitrate. Rates of denitrification were stimulated to a larger extent by enhanced nitrite than by nitrate concentrations. At 550 microM nitrite or nitrate (the highest concentration used), the rates of denitrification were 600 micromol N.m(-2).h(-1) with nitrite but only 180 micromol N.m(-2).h(-1) with nitrate. The ratios of rates of nitrous oxide production and denitrification (N(2)O/N(2) x 100) were significantly higher with the addition of nitrite (7 to 13% of denitrification) than with nitrate (2 to 4% of denitrification). The results suggested that in addition to anaerobic bacteria, which possess the complete denitrification pathway for N(2) formation in the estuarine sediments, there may be two other groups of bacteria: nitrite denitrifiers, which reduce nitrite to N(2) via N(2)O, and obligate nitrite-denitrifying bacteria, which reduce nitrite to N(2)O as the end product. Consideration of free-energy changes during N(2)O formation led to the conclusion that N(2)O formation using nitrite as the electron acceptor is favored in the Colne estuary and may be a critical factor regulating the formation of N(2)O in high-nutrient-load estuaries. PMID- 11872475 TI - Novel alpha-glucosidase from Aspergillus nidulans with strong transglycosylation activity. AB - Aspergillus nidulans possessed an alpha-glucosidase with strong transglycosylation activity. The enzyme, designated alpha-glucosidase B (AgdB), was purified and characterized. AgdB was a heterodimeric protein comprising 74- and 55-kDa subunits and catalyzed hydrolysis of maltose along with formation of isomaltose and panose. Approximately 50% of maltose was converted to isomaltose, panose, and other minor transglycosylation products by AgdB, even at low maltose concentrations. The agdB gene was cloned and sequenced. The gene comprised 3,055 bp, interrupted by three short introns, and encoded a polypeptide of 955 amino acids. The deduced amino acid sequence contained the chemically determined N terminal and internal amino acid sequences of the 74- and 55-kDa subunits. This implies that AgdB is synthesized as a single polypeptide precursor. AgdB showed low but overall sequence homology to alpha-glucosidases of glycosyl hydrolase family 31. However, AgdB was phylogenetically distinct from any other alpha glucosidases. We propose here that AgdB is a novel alpha-glucosidase with unusually strong transglycosylation activity. PMID- 11872476 TI - Genetic relationship among worldwide strains of Xanthomonas causing canker in citrus species and design of new primers for their identification by PCR. AB - Partial sequence analysis of the ribosomal operon in Xanthomonas axonopodis allowed discrimination among strains causing the A, B, and C types of citrus bacterial canker (CBC) and quantification of the relationship of these organisms with other species and pathovars in the same genus. Sets of primers based on sequence differences in the internally transcribed spacer and on a sequence from the plasmid gene pthA involved in virulence were designed for specific identification of xanthomonads causing CBC diseases. The two sets were validated with a collection of Xanthomonas strains associated with citrus species. The primer set based on ribosomal sequences had a high level of specificity for X. axonopodis pv. citri, whereas the set based on the pthA gene was universal for all types of CBC organisms. Moreover, the relationships among worldwide Xanthomonas strains causing CBC were analyzed by amplification of repetitive sequences (enterobacterial repetitive intergenic consensus and BOX elements). Under specific conditions, pathotypes of these Xanthomonas strains could be discerned, and subgroups of the pathotypes were identified. Subgroups of strains were associated with certain geographic areas of the world, and on this basis the origin of type A strains introduced into Florida could be inferred. PMID- 11872478 TI - Reversal of flagellar rotation is important in initial attachment of Escherichia coli to glass in a dynamic system with high- and low-ionic-strength buffers. AB - The attachment rates of wild-type, smooth-swimming, tumbly, and paralyzed Escherichia coli to glass was measured at fluid velocities of 0.0044 and 0.044 cms(-1) (corresponding to shear rates of 0.34 and 3.4 s(-1), respectively), in 0.02 and 0.2 M buffer solutions. At the highest ionic strength, we did not observe a significant difference in the attachment rate of wild-type and paralyzed cells at either fluid velocity. However, when the ionic strength was reduced, paralyzed bacteria attached at rates 4 and 10 times lower than that of the wild type under fluid velocities of 0.0044 and 0.044 cms(-1), respectively. This suggested that the rotation of the flagella assisted in attachment. We then compared the attachment rates of smooth-swimming (counterclockwise rotation only) and tumbly (clockwise rotation only) cells to the wild type to determine whether the direction of rotation was important to cell attachment. At 0.0044 cms(-1), the smooth-swimming cells attached at rates similar to that of the wild type in both buffer solutions but significantly less at the higher fluid velocity. Tumbly cells attached at much lower rates under all conditions. Thus, the combination of clockwise and counterclockwise flagellar rotation and their coupling appeared to be important in cell attachment. We considered a number of hypotheses to interpret these observations, including a residence time analysis and a comparison of traditional Derjaguin-Landau-Verwey-Overbeek (DLVO) theory to soft particle theory. PMID- 11872477 TI - Molecular and culture-based analyses of prokaryotic communities from an agricultural soil and the burrows and casts of the earthworm Lumbricus rubellus. AB - The microbial populations in no-till agricultural soil and casts of the earthworm Lumbricus rubellus were examined by culturing and molecular methods. Clone libraries of the 16S rRNA genes were prepared from DNA isolated directly from the soil and earthworm casts. Although no single phylum dominated the soil library of 95 clones, the largest numbers of clones were from Acidobacteria (14%), Cytophagales (13%), Chloroflexi (8%), and gamma-Proteobacteria (8%). While the cast clone library of 102 clones was similar to the soil library, the abundances of several taxa were different. Representatives of the Pseudomonas genus as well as the Actinobacteria and Firmicutes increased in number, and one group of unclassified organisms found in the soil library was absent in the cast library. Likewise, soil and cast archaeal 16S rRNA gene libraries were similar, although the abundances of some groups were different. Two hundred and thirty aerobic bacteria were also isolated on general heterotrophic media from casts, burrows, and soil. The cast isolates were both phenotypically and genotypically different from the soil isolates. The cast isolates were more likely to reduce nitrate, grow on acetate and Casamino Acids, and utilize fewer sugars than the soil isolates. On the basis of their ribotypes, the cast isolates were dominated by Aeromonas spp. (28%), which were not found in the soil isolates, and other gamma Proteobacteria (49%). In contrast, the soil isolates were mostly Actinobacteria (53%), Firmicutes (16%), and gamma-Proteobacteria (19%). Isolates obtained from the sides of earthworm burrows were not different from the soil isolates. Diversity indices for the collections of isolates as well as rRNA gene libraries indicated that the species richness and evenness were decreased in the casts from their levels in the soil. These results were consistent with a model where a large portion of the microbial population in soil passes through the gastrointestinal tract of the earthworm unchanged while representatives of some phyla increase in abundance. PMID- 11872479 TI - Sequence analysis of marine virus communities reveals that groups of related algal viruses are widely distributed in nature. AB - Algal-virus-specific PCR primers were used to amplify DNA polymerase (pol) gene fragments from geographically isolated natural virus communities. Natural algal virus communities were obtained from coastal sites in the Pacific Ocean in British Columbia, Canada, and the Southern Ocean near the Antarctic peninsula. Genetic fingerprints of algal virus communities were generated using denaturing gradient gel electrophoresis (DGGE). Sequencing efforts recovered 33 sequences from the gradient gel. Of the 33 sequences examined, 25 encoded a conserved amino acid motif indicating that the sequences were pol gene fragments. Furthermore, the 25 pol sequences were related to pol gene fragments from known algal viruses. In addition, similar virus sequences (>98% sequence identity) were recovered from British Columbia and Antarctica. Results from this study demonstrate that DGGE with degenerate primers can be used to qualitatively fingerprint and assess genetic diversity in specific subsets of natural virus communities and that closely related viruses occur in distant geographic locations. DGGE is a powerful tool for genetically fingerprinting natural virus communities and may be used to examine how specific components of virus communities respond to experimental manipulations. PMID- 11872480 TI - Purification and characterization of an extracellular protease from Xenorhabdus nematophila involved in insect immunosuppression. AB - Xenorhabdus nematophila, a bacterium pathogenic for insects associated with the nematode Steinernema carpocapsae, releases high quantities of proteases, which may participate in the virulence against insects. Zymogram assays and cross reactions of antibodies suggested that two distinct proteases were present. The major one, protease II, was purified and shown to have a molecular mass of 60 kDa and an estimated isoelectric point of 8.5. Protease II digested the chromogenic substrate N-tosyl-Gly-Pro-Arg-paranitroanilide (pNA) with V(max) and K(m) values of 0.0551 microM/min and 234 microM, respectively, and the substrate DL-Val-Leu Arg-pNA with V(max) and K(m) values of 0.3830 microM/min and 429 microM, respectively. Protease II activity was inhibited 93% by Pefabloc SC and 45% by chymostatin. The optimum pH for protease II was 7, and the optimum temperature was 23C. Proteolytic activity was reduced by 90% at 60 degrees C for 10 min. Sequence analysis was performed on four internal peptides that resulted from the digestion of protease II. Fragments 29 and 45 are 75 and 68% identical to alkaline metalloproteinase produced by Pseudomonas aeruginosa. Fragment 29 is 79% identical to a metalloprotease of Erwinia amylovora and 75% identical to the protease C precursor of Erwinia chrysanthemi. Protease II showed no toxicity to hemocytes but destroyed antibacterial activity on the hemolymph of inoculated insects' larvae and reduced 97% of the cecropin A bacteriolytic activity. PMID- 11872482 TI - Production of N(2) through anaerobic ammonium oxidation coupled to nitrate reduction in marine sediments. AB - In the global nitrogen cycle, bacterial denitrification is recognized as the only quantitatively important process that converts fixed nitrogen to atmospheric nitrogen gas, N(2), thereby influencing many aspects of ecosystem function and global biogeochemistry. However, we have found that a process novel to the marine nitrogen cycle, anaerobic oxidation of ammonium coupled to nitrate reduction, contributes substantially to N(2) production in marine sediments. Incubations with (15)N-labeled nitrate or ammonium demonstrated that during this process, N(2) is formed through one-to-one pairing of nitrogen from nitrate and ammonium, which clearly separates the process from denitrification. Nitrite, which accumulated transiently, was likely the oxidant for ammonium, and the process is thus similar to the anammox process known from wastewater bioreactors. Anaerobic ammonium oxidation accounted for 24 and 67% of the total N(2) production at two typical continental shelf sites, whereas it was detectable but insignificant relative to denitrification in a eutrophic coastal bay. However, rates of anaerobic ammonium oxidation were higher in the coastal sediment than at the deepest site and the variability in the relative contribution to N(2) production between sites was related to large differences in rates of denitrification. Thus, the relative importance of anaerobic ammonium oxidation and denitrification in N(2) production appears to be regulated by the availability of their reduced substrates. By shunting nitrogen directly from ammonium to N(2), anaerobic ammonium oxidation promotes the removal of fixed nitrogen in the oceans. The process can explain ammonium deficiencies in anoxic waters and sediments, and it may contribute significantly to oceanic nitrogen budgets. PMID- 11872481 TI - Cloning, characterization, and transcription of three laccase genes from Gaeumannomyces graminis var. tritici, the take-all fungus. AB - Gaeumannomyces graminis var. tritici, a filamentous ascomycete, is an important root pathogen of cereals that causes take-all disease and results in severe crop losses worldwide. Previously we identified a polyphenol oxidase (laccase) secreted by the fungus when induced with copper. Here we report cloning and partial characterization of three laccase genes (LAC1, LAC2, and LAC3) from G. graminis var. tritici. Predicted polypeptides encoded by these genes had 38 to 42% amino acid sequence identity and had conserved copper-binding sites characteristic of laccases. The sequence of the LAC2 predicted polypeptide matched the N-terminal sequence of the secreted laccase that we purified in earlier studies. We also characterized expression patterns of these genes by reverse transcription-PCR. LAC1 was transcribed constitutively, and transcription of LAC2 was Cu inducible. All three genes were transcribed in planta; however, transcription of LAC3 was observed only in planta or in the presence of host (wheat) plant homogenate. PMID- 11872483 TI - Detection of Campylobacter jejuni and Campylobacter coli in environmental waters by PCR enzyme-linked immunosorbent assay. AB - A PCR enzyme-linked immunosorbent assay (ELISA) assay was applied to the detection of Campylobacter jejuni and Campylobacter coli in environmental water samples after enrichment culture. Bacterial cells were concentrated from 69 environmental water samples by using filtration, and the filtrates were cultured in Campylobacter blood-free broth. After enrichment culture, DNA was extracted from the samples by using a rapid-boiling method, and the DNA extracts were used as a template in a PCR ELISA assay. A total of 51 samples were positive by either PCR ELISA or culture; of these, 43 were found to be positive by PCR ELISA and 43 were found to be positive by culture. Overall, including positive and negative results, 59 samples were concordant in both methods. Several samples were positive in the PCR ELISA assay but were culture negative; therefore, this assay may be able to detect sublethally damaged or viable nonculturable forms of campylobacters. The method is rapid and sensitive, and it significantly reduces the time needed for the detection of these important pathogens by 2 to 3 days. PMID- 11872484 TI - Effects of T4 lysozyme release from transgenic potato roots on bacterial rhizosphere communities are negligible relative to natural factors. AB - Rhizosphere bacterial communities of two transgenic potato lines which produce T4 lysozyme for protection against bacterial infections were analyzed in comparison to communities of wild-type plants and transgenic controls not harboring the lysozyme gene. Rhizosphere samples were taken from young, flowering, and senescent plants at two field sites in three consecutive years. The communities were characterized in a polyphasic approach. Cultivation-dependent methods included heterotrophic plate counts, determination of species composition and diversity based on fatty acid analysis of isolates, and community level catabolic profiling. Cultivation-independent analyses were based on denaturing gradient gel electrophoresis (DGGE) of 16S rRNA gene fragments amplified from rhizosphere DNA using primers specific for Bacteria, Actinomycetales, or alpha- or beta Proteobacteria. Several bands of the DGGE patterns were further characterized by sequence analysis. All methods revealed that environmental factors related to season, field site, or year but not to the T4 lysozyme expression of the transgenic plants influenced the rhizosphere communities. For one of the T4 lysozyme-producing cultivars, no deviation in the rhizosphere communities compared to the control lines was observed. For the other, differences were detected at some of the samplings between the rhizosphere community structure and those of one or all other cultivars which were not attributable to T4 lysozyme production but most likely to differences observed in the growth characteristics of this cultivar. PMID- 11872486 TI - Production of new unsaturated lipids during wood decay by ligninolytic basidiomycetes. AB - Lipids were analyzed by gas chromatography-mass spectrometry for a 7-week in vitro decay of eucalypt wood by four ligninolytic basidiomycetes. The sound wood contained up to 75 mg of lipophilic compounds per 100 g of wood. Hydrolysis of sterol esters, which represented 38% of total wood lipids, occurred during the fungal decay. The initial increase of linoleic and other free unsaturated fatty acids paralleled the decrease of sterol esters. Moreover, new lipid compounds were found at advanced stages of wood decay that were identified from their mass spectra as unsaturated dicarboxylic acids consisting of a long aliphatic chain attached to the C-3 position of itaconic acid. These dicarboxylic acids were especially abundant in the wood treated with Ceriporiopsis subvermispora (up to 24 mg per 100 g of wood) but also were produced by Phlebia radiata, Pleurotus pulmonarius, and Bjerkandera adusta. We hypothesize that three main alkylitaconic acids (tetradecylitaconic, cis-7-hexadecenylitaconic, and hexadecylitaconic acids) are synthesized by fungi in condensation reactions involving palmitic, oleic, and stearic acids. We suggest that both wood unsaturated fatty acids (present in free form or released from esters during natural decay) and unsaturated metabolites synthesized by fungi could serve as a source for peroxidizable lipids in lignin degradation by white rot basidiomycetes. PMID- 11872487 TI - Quantitative analysis of the relative transcript levels of ABC transporter Atr genes in Aspergillus nidulans by real-time reverse transcription-PCR assay. AB - The development of assays for quantitative analysis of the relative transcript levels of ABC transporter genes by real-time reverse transcription-PCR (RT-PCR) might provide important information about multidrug resistance in filamentous fungi. Here, we evaluate the potential of real-time RT-PCR to quantify the relative transcript levels of ABC transporter Atr genes from Aspergillus nidulans. The AtrA to AtrD genes showed different and higher levels in the presence of structurally unrelated drugs, such as camptothecin, imazalil, itraconazole, hygromycin, and 4-nitroquinoline oxide. We also verified the relative transcript levels of the Atr genes in the A. nidulans imazalil-resistant mutants. These genes displayed a very complex pattern in different ima genetic backgrounds. The imaB mutant has higher basal transcript levels of AtrB and -D than those of the wild-type strain. The levels of these two genes are comparable when the imaB mutant is grown in the presence and absence of imazalil. The imaC, D, and -H mutants have higher basal levels of AtrA than that of the wild type. The same behavior is observed for the relative transcript levels of AtrB in the imaG mutant background. PMID- 11872485 TI - Saccharomyces cerevisiae URH1 (encoding uridine-cytidine N-ribohydrolase): functional complementation by a nucleoside hydrolase from a protozoan parasite and by a mammalian uridine phosphorylase. AB - Nucleoside hydrolases catalyze the cleavage of N-glycosidic bonds in nucleosides, yielding ribose and the respective bases. While nucleoside hydrolase activity has not been detected in mammalian cells, many protozoan parasites rely on nucleoside hydrolase activity for salvage of purines and/or pyrimidines from their hosts. In contrast, uridine phosphorylase is the key enzyme of pyrimidine salvage in mammalian hosts and many other organisms. We show here that the open reading frame (ORF) YDR400w of Saccharomyces cerevisiae carries the gene encoding uridine hydrolase (URH1). Disruption of this gene in a conditionally pyrimidine auxotrophic S. cerevisiae strain, which is also deficient in uridine kinase (urk1), leads to the inability of the mutant to utilize uridine as the sole source of pyrimidines. Protein extracts of strains overexpressing YDR400w show increased hydrolase activity only with uridine and cytidine, but no activity with inosine, adenosine, guanosine, and thymidine as substrates, demonstrating that ORF YDR400w encodes a uridine-cytidine N-ribohydrolase. Expression of a homologous cDNA from a protozoan parasite (Crithidia fasciculata) in a ura3 urk1 urh1 mutant is sufficient to restore growth on uridine. Growth can also be restored by expression of a human uridine phosphorylase cDNA. Yeast strains expressing protozoan N-ribohydrolases or host phosphorylases could therefore become useful tools in drug screens for specific inhibitors. PMID- 11872488 TI - Novel psbA1 gene from a naturally occurring atrazine-resistant cyanobacterial isolate. AB - A naturally occurring atrazine-resistant cyanobacterial isolate, strain SG2, was isolated from an atrazine-containing wastewater treatment system at the Syngenta atrazine production facility in St. Gabriel, La. Strain SG2 was resistant to 1,000 microg of atrazine per ml but showed relatively low resistance to diuron [3 (3,4-dichlorophenyl)-1,1-dimethyl urea]. Analyses of 16S ribosomal DNA indicated that strain SG2 falls into the Synechocystis/Pleurocapsa/Microcystis group. Photosynthetically driven oxygen evolution in strain SG2 was only slightly inhibited (about 10%) by 2,000 microg of atrazine per ml, whereas in the control strain Synechocystis 6803, oxygen evolution was inhibited 90% by 1,000 microg of atrazine per ml. No atrazine accretion, mineralization, or metabolites were detected when strain SG2 was grown with [(14)C]atrazine. Strain SG2 contained three copies of the psbA gene, which encodes the D(1) protein of the photosystem II reaction center. Nucleotide sequence analyses indicated that the psbA2 and psbA3 genes encoded predicted proteins with the same amino acid sequence. However, the psbA1 gene product contained five extra amino acids, which were not found in PsbA proteins from five other cyanobacteria. Moreover, the PsbA1 protein from strain SG2 had an additional 13 amino acid changes compared to the PsbA2/PsbA3 proteins and contained 10 amino acid alterations compared to conserved residues found in other cyanobacteria. Reverse transcriptase PCR analysis indicated that the psbA1 gene and the psbA2/psbA3 gene(s) were expressed in photosynthetically grown cells in the presence of atrazine. These results suggest that strong selection pressure conferred by the continual input of atrazine has contributed to the evolution of a herbicide-resistant, yet photosynthetically efficient, psbA gene in a cyanobacterium. PMID- 11872489 TI - Surface modifications created by using engineered hydrophobins. AB - Hydrophobins are small (ca. 100 amino acids) secreted fungal proteins that are characterized by the presence of eight conserved cysteine residues and by a typical hydropathy pattern. Class I hydrophobins self-assemble at hydrophilic hydrophobic interfaces into highly insoluble amphipathic membranes, thereby changing the nature of surfaces. Hydrophobic surfaces become hydrophilic, while hydrophilic surfaces become hydrophobic. To see whether surface properties of assembled hydrophobins can be changed, 25 N-terminal residues of the mature SC3 hydrophobin were deleted (TrSC3). In addition, the cell-binding domain of fibronectin (RGD) was fused to the N terminus of mature SC3 (RGD-SC3) and TrSC3 (RGD-TrSC3). Self-assembly and surface activity were not affected by these modifications. However, physiochemical properties at the hydrophilic side of the assembled hydrophobin did change. This was demonstrated by a change in wettability and by enhanced growth of fibroblasts on Teflon-coated with RGD-SC3, TrSC3, or RGD-TrSC3 compared to bare Teflon or Teflon coated with SC3. Thus, engineered hydrophobins can be used to functionalize surfaces. PMID- 11872490 TI - Hindgut fermentation in three species of marine herbivorous fish. AB - Symbioses with gut microorganisms provides a means by which terrestrial herbivores are able to obtain energy. These microorganisms ferment cell wall materials of plants to short-chain fatty acids (SCFA), which are then absorbed and used by the host animal. Many marine herbivorous fishes contain SCFA (predominantly acetate) in their hindgut, indicative of gut microbial activity, but rates of SCFA production have not been measured. Such information is an important prerequisite to understanding the contribution that gut microorganisms make in satisfying the energy needs of the fish. We have estimated the rates of acetate production in the gut of three species of temperate marine herbivorous fish from northeastern New Zealand: Kyphosus sydneyanus (family Kyphosidae), Odax pullus (family Odacidae), and Aplodactylus arctidens (family Aplodactylidae). Ex vivo preparations of freshly caught fish were maintained with their respiratory and circulatory systems intact, radiolabeled acetate was injected into ligated hindgut sections, and gut fluid was sampled at 20-min intervals for 2 h. Ranges for acetate turnover in the hindguts of the studied species were determined from the slope of plots as the log of the specific radioactivity of acetate versus time and pool size, expressed on a nanomole per milliliter per minute basis. Values were 450 to 570 (K. sydneyanus), 373 to 551 (O. pullus), and 130 to 312 (A. arctidens). These rates are comparable to those found in the guts of herbivorous reptiles and mammals. To determine the contribution of metabolic pathways to the fate of acetate, rates of sulfate reduction and methanogenesis were measured in the fore-, mid-, and hindgut sections of the three fish species. Both rates increased from the distal to proximal end of the hindgut, where sulfate reduction accounted for only a small proportion (<5%) of acetate methyl group transformed to CO(2), and exceeded methanogenesis from acetate by >50-fold. When gut size was taken into account, acetate uptake from the hindgut of the fish species, determined on a millimole per day per kilogram of body weight basis, was 70 (K. sydneyanus), 18 (O. pullus), and 10 (A. arctidens). PMID- 11872491 TI - Intraspecies genomic groups in Enterococcus faecium and their correlation with origin and pathogenicity. AB - Seventy-eight Enterococcus faecium strains from various sources were characterized by random amplified polymorphic DNA (RAPD)-PCR, amplified fragment length polymorphism (AFLP), and pulsed-field gel electrophoresis (PFGE) analysis of SmaI restriction patterns. Two main genomic groups (I and II) were obtained in both RAPD-PCR and AFLP analyses. DNA-DNA hybridization values between representative strains of both groups demonstrated a mean DNA-DNA reassociation level of 71%. PFGE analysis revealed high genetic strain diversity within the two genomic groups. Only group I contained strains originating from human clinical samples or strains that were vancomycin-resistant or beta-hemolytic. No differentiating phenotypic features between groups I and II were found using the rapid ID 32 STREP system. The two groups could be further subdivided into, respectively, four and three subclusters in both RAPD-PCR and AFLP analyses, and a high correlation was seen between the subclusters generated by these two methods. Subclusters of group I were to some extent correlated with origin, pathogenicity, and bacteriocinogeny of the strains. Host specificity of E. faecium strains was not confirmed. PMID- 11872492 TI - Successional development of sulfate-reducing bacterial populations and their activities in a wastewater biofilm growing under microaerophilic conditions. AB - A combination of fluorescence in situ hybridization, microprofiles, denaturing gradient gel electrophoresis of PCR-amplified 16S ribosomal DNA fragments, and 16S rRNA gene cloning analysis was applied to investigate successional development of sulfate-reducing bacteria (SRB) community structure and in situ sulfide production activity within a biofilm growing under microaerophilic conditions (dissolved oxygen concentration in the bulk liquid was in the range of 0 to 100 microM) and in the presence of nitrate. Microelectrode measurements showed that oxygen penetrated 200 microm from the surface during all stages of biofilm development. The first sulfide production of 0.32 micromol of H(2)S m(-2) s(-1) was detected below ca. 500 microm in the 3rd week and then gradually increased to 0.70 micromol H(2)S m(-2) s(-1) in the 8th week. The most active sulfide production zone moved upward to the oxic-anoxic interface and intensified with time. This result coincided with an increase in SRB populations in the surface layer of the biofilm. The numbers of the probe SRB385- and 660-hybridized SRB populations significantly increased to 7.9 x 10(9) cells cm(-3) and 3.6 x 10(9) cells cm(-3), respectively, in the surface 400 microm during an 8-week cultivation, while those populations were relatively unchanged in the deeper part of the biofilm, probably due to substrate transport limitation. Based on 16S rRNA gene cloning analysis data, clone sequences that related to Desulfomicrobium hypogeium (99% sequence similarity) and Desulfobulbus elongatus (95% sequence similarity) were most frequently found. Different molecular analyses confirmed that Desulfobulbus, Desulfovibrio, and Desulfomicrobium were found to be the numerically important members of SRB in this wastewater biofilm. PMID- 11872493 TI - Reduction of olive knot disease by a bacteriocin from Pseudomonas syringae pv. ciccaronei. AB - A bacteriocin produced by Pseudomonas syringae pv. ciccaronei, used at different purification levels and concentrations in culture and in planta, inhibited the multiplication of P. syringae subsp. savastanoi, the causal agent of olive knot disease, and affected the epiphytic survival of the pathogen on the leaves and twigs of treated olive plants. Treatments with bacteriocin from P. syringae pv. ciccaronei inhibited the formation of overgrowths on olive plants caused by P. syringae subsp. savastanoi strains PVBa229 and PVBa304 inoculated on V-shaped slits and on leaf scars at concentrations of 10(5) and 10(8) CFU ml(-1), respectively. In particular, the application of 6,000 arbitrary units (AU) of crude bacteriocin (dialyzed ammonium sulfate precipitate of culture supernatant) ml(-1) at the inoculated V-shaped slits and leaf scars resulted in the formation of knots with weight values reduced by 81 and 51%, respectively, compared to the control, depending on the strains and inoculation method used. Crude bacteriocin (6,000 AU ml(-1)) was also effective in controlling the multiplication of epiphytic populations of the pathogen. In particular, the bacterial populations recovered after 30 days were at least 350 and 20 times lower than the control populations on twigs and on leaves, respectively. These results suggest that bacteriocin from P. syringae pv. ciccaronei can be used effectively to control the survival of the causal agent of olive knot disease and to prevent its multiplication at inoculation sites. PMID- 11872494 TI - Identification of a hydrophobin gene that is developmentally regulated in the ectomycorrhizal fungus Tricholoma terreum. AB - The symbiosis between ectomycorrhizal fungi and trees is an essential part of forest ecology and depends entirely on the communication between the two partners for establishing and maintaining the relationship. The identification and characterization of differentially expressed genes is a step to identifying such signals and to understanding the regulation of this process. We determined the role of hydrophobins produced by Tricholoma terreum in mycorrhiza formation and hyphal development. A hydrophobin was purified from culture supernatant, and the corresponding gene was identified. The gene is expressed in aerial mycelium and in mycorrhiza. By using a heterologous antiserum directed against a hydrophobin found in the aerial mycelium of Schizophyllum commune, we detected a hydrophobin in the symbiosis between T. terreum and its native pine host Pinus sylvestris. The hydrophobin was found in aerial mycelium of the hyphal mantle and also in the Hartig net hyphae, which form the interface between both partners. Interestingly, this was not the case in the interaction of T. terreum with a host of low compatibility, the spruce Picea abies. The differential expression with respect to host was verified at the transcriptional level by competitive PCR. The differential protein accumulation pattern with respect to host compatibility seen by immunofluorescence staining can thus be attributed at least in part to transcriptional control of the hyd1 gene. PMID- 11872495 TI - Genetic differentiation between sympatric populations of Bacillus cereus and Bacillus thuringiensis. AB - Little is known about genetic exchanges in natural populations of bacteria of the spore-forming Bacillus cereus group, because no population genetics studies have been performed with local sympatric populations. We isolated strains of Bacillus thuringiensis and B. cereus from small samples of soil collected at the same time from two separate geographical sites, one within the forest and the other at the edge of the forest. A total of 100 B. cereus and 98 B. thuringiensis strains were isolated and characterized by electrophoresis to determine allelic composition at nine enzymatic loci. We observed genetic differentiation between populations of B. cereus and B. thuringiensis. Populations of a given Bacillus species--B. thuringiensis or B. cereus--were genetically more similar to each other than to populations of the other Bacillus species. Hemolytic activity provided further evidence of this genetic divergence, which remained evident even if putative clones were removed from the data set. Our results suggest that the rate of gene flow was higher between strains of the same species, but that exchanges between B. cereus and B. thuringiensis were nonetheless possible. Linkage disequilibrium analysis revealed sufficient recombination for B. cereus populations to be considered panmictic units. In B. thuringiensis, the balance between clonal proliferation and recombination seemed to depend on location. Overall, our data indicate that it is not important for risk assessment purposes to determine whether B. cereus and B. thuringiensis belong to a single or two species. Assessment of the biosafety of pest control based on B. thuringiensis requires evaluation of the extent of genetic exchange between strains in realistic natural conditions. PMID- 11872496 TI - Quantitative detection of microbial genes by using DNA microarrays. AB - To quantify target genes in biological samples using DNA microarrays, we employed reference DNA to normalize variations in spot size and hybridization. This method was tested using nitrate reductase (nirS), naphthalene dioxygenase (nahA), and Escherichia coli O157 O-antigen biosynthesis genes as model genes and lambda DNA as the reference DNA. We observed a good linearity between the log signal ratio and log DNA concentration ratio at DNA concentrations above the method's detection limit, which was approximately 10 pg. This approach for designing quantitative microarrays and the inferred equation from this study provide a simple and convenient way to estimate the target gene concentration from the hybridization signal ratio. PMID- 11872497 TI - Modeling growth and bacteriocin production by Lactobacillus amylovorus DCE 471 in response to temperature and pH values used for sourdough fermentations. AB - The biokinetics of cell growth of Lactobacillus amylovorus DCE 471 and bacteriocin production by this strain were investigated as a function of the temperatures (28 to 44C) and pH values (pH 4.2 to 6.4) that are characteristic of a sourdough fermentation process. The influence of temperature and pH on microbial behavior is described by using a successfully validated predictive model. PMID- 11872498 TI - Isolation and genotyping of Helicobacter pylori from untreated municipal wastewater. AB - For this study, we isolated Helicobacter pylori from wastewater by a series of steps beginning with immunomagnetic separation and cell culture. After Gram staining and three standard microbial tests, the 16S rRNA sequences of a total of 23 out of 37 putative H. pylori isolates were verified by PCR. Eleven H. pylori isolates were genotyped and fell into four vacA classes: those with the vacA allelic variants s1a and m1, s1b and m1, s2 and m2, or s2 and m1. Most H. pylori isolates were of the vacA s1a/m1 type, which has been shown to be associated with advanced diseases based on genotyping of H. pylori from gastric cancer patients. These results demonstrated that H. pylori survives in water and may be a potential source of H. pylori transmission, especially where water is not adequately treated. PMID- 11872499 TI - Microbial iron respiration can protect steel from corrosion. AB - Microbiologically influenced corrosion (MC) of steel has been attributed to the activity of biofilms that include anaerobic microorganisms such as iron-respiring bacteria, yet the mechanisms by which these organisms influence corrosion have been unclear. To study this process, we generated mutants of the iron-respiring bacterium Shewanella oneidensis strain MR-1 that were defective in biofilm formation and/or iron reduction. Electrochemical impedance spectroscopy was used to determine changes in the corrosion rate and corrosion potential as a function of time for these mutants in comparison to the wild type. Counter to prevailing theories of MC, our results indicate that biofilms comprising iron-respiring bacteria may reduce rather than accelerate the corrosion rate of steel. Corrosion inhibition appears to be due to reduction of ferric ions to ferrous ions and increased consumption of oxygen, both of which are direct consequences of microbial respiration. PMID- 11872501 TI - Nitrite as a stimulus for ammonia-starved Nitrosomonas europaea. AB - Ammonia-starved cells of Nitrosomonas europaea are able to preserve a high level of ammonia-oxidizing activity in the absence of ammonium. However, when the nitrite-oxidizing cells that form part of the natural nitrifying community do not keep pace with the ammonia-oxidizing cells, nitrite accumulates and may subsequently inhibit ammonia oxidation. The maintenance of a high ammonia oxidizing capacity during starvation is then nullified. In this study we demonstrated that cells of N. europaea starved for ammonia were not sensitive to nitrite, either when they were starved in the presence of nitrite or when nitrite was supplied simultaneously with fresh ammonium. In the latter case, the initial ammonia-oxidizing activity of starved cells was stimulated at least fivefold. PMID- 11872500 TI - Identification of the functionally active methanotroph population in a peat soil microcosm by stable-isotope probing. AB - The active population of low-affinity methanotrophs in a peat soil microcosm was characterized by stable-isotope probing. "Heavy" (13)C-labeled DNA, produced after microbial growth on (13)CH(4), was separated from naturally abundant (12)C DNA by cesium chloride density gradient centrifugation and used as a template for the PCR. Amplification products of 16S rRNA genes and pmoA, mxaF, and mmoX, which encode key enzymes in the CH(4) oxidation pathway, were analyzed. Sequences related to extant type I and type II methanotrophs were identified, indicating that these methanotrophs were active in peat exposed to 8% (vol/vol) CH(4). The (13)C-DNA libraries also contained clones that were related to beta-subclass Proteobacteria, suggesting that novel groups of bacteria may also be involved in CH(4) cycling in this soil. PMID- 11872502 TI - Rupture of the cell envelope by decompression of the deep-sea methanogen Methanococcus jannaschii. AB - The effect of decompression on the structure of Methanococcus jannaschii, an extremely thermophilic deep-sea methanogen, was studied in a novel high-pressure, high-temperature bioreactor. The cell envelope of M. jannaschii appeared to rupture upon rapid decompression (ca. 1 s) from 260 atm of hyperbaric pressure. When decompression from 260 atm was performed over 5 min, the proportion of ruptured cells decreased significantly. In contrast to the effect produced by decompression from hyperbaric pressure, decompression from a hydrostatic pressure of 260 atm did not induce cell lysis. PMID- 11872503 TI - Continuous steady-state method using tenax for delivering tetrachloroethene to chloro-respiring bacteria. AB - Tenax-TA, a solid-phase sorbent, was used as an alternative to hexadecane for continuous delivery of tetrachloroethene (PCE) to Desulfuromonas strain BB1, a chloro-respiring microorganism. In both batch and bioreactor configurations, Tenax not only maintained low, steady-state concentrations of PCE in an active culture for several months but also adsorbed the product of dechlorination, cis 1,2-dichloroethene, before it approached toxic levels. PMID- 11872504 TI - Higher abundance of bacteria than of viruses in deep Mediterranean sediments. AB - The interactions between viral abundance and bacterial density, biomass, and production were investigated along a longitudinal transect consisting of nine deep-sea stations encompassing the entire Mediterranean basin. The numbers of viruses were very low (range, 3.6 x 10(7) to 12.0 x 10(7) viruses g(-1)) and decreased eastward. The virus-to-bacterium ratio was always < 1.0, indicating that the deep-sea sediments of the Mediterranean Sea are the first example of a marine ecosystem not numerically dominated by viruses. The lowest virus numbers were found where the lowest bacterial metabolism and turnover rates and the largest cell size were observed, suggesting that bacterial doubling time might play an important role in benthic virus development. PMID- 11872505 TI - Inhibition of bacterial growth, enterotoxin production, and spore outgrowth in strains of Bacillus cereus by bacteriocin AS-48. AB - Bacteriocin AS-48 showed high bactericidal activity for mesophilic and psychrotrophic strains of Bacillus cereus over a broad pH range. AS-48 inhibition of the enterotoxin-producing strain LWL1 was enhanced by sodium nitrite, sodium lactate, and sodium chloride. The latter also enhanced AS-48 activity against strain CECT 131. Bacterial growth and enterotoxin production by strain LWL1 were completely inhibited at bacteriocin concentrations of 7.5 microg/ml. At subinhibitory bacteriocin concentrations, enterotoxin production decreased markedly and sporulation was delayed. Intact spores were resistant to AS-48 but became gradually sensitive to AS-48 during the course of germination. PMID- 11872507 TI - The antidepressant debate. PMID- 11872506 TI - Widespread distribution in polar oceans of a 16S rRNA gene sequence with affinity to Nitrosospira-like ammonia-oxidizing bacteria. AB - We analyzed the phylogenetic compositions of ammonia-oxidizing bacteria of the beta subclass of Proteobacteria from 42 Southern Ocean samples. We found a Nitrosospira-like 16S rRNA gene sequence in all 20 samples that yielded PCR products (8 of 30 samples from the Ross Sea and 12 of 12 samples from the Palmer Peninsula). We also found this sequence in Arctic Ocean samples, indicating a transpolar, if not global, distribution; however, slight differences between Arctic and Antarctic sequences may be evidence of polar endemism. PMID- 11872508 TI - Fear knot. Neurobiological disruption of long-term fear memory. PMID- 11872509 TI - Ethnic minorities and the Mental Health Act. PMID- 11872510 TI - The maturing of therapy. Some brief psychotherapies help anxiety/depressive disorders but mechanisms of action are unclear. AB - BACKGROUND: Psychiatric therapy needs assessment regarding its maturation as a therapeutic science. AIMS: Judgement of whether such a science is emerging. METHOD: Four criteria are used: efficacy; identification of responsible treatment components; knowledge of their mechanisms of action; and elucidation of why they act only in some sufferers. RESULTS: Brief behavioural, interpersonal, cognitive, problem-solving and other psychotherapies have a mature ability to improve anxiety and depressive disorders reliably and enduringly, often only with instruction from a manual or a computer. Therapy's cost-effectiveness and acceptability deserve more attention. We know little about which treatment components produce improvement, how they do so and why they do not help all sufferers. CONCLUSIONS: Therapy is coming of age regarding efficacy for anxiety and depression, but is only a toddler regarding the scientific principles to explain its effects. PMID- 11872511 TI - Impact of Event Scale: psychometric properties. AB - BACKGROUND: For more than 20 years, the Impact of Event Scale (IES) has been widely used as a measure of stress reactions after traumatic events. AIMS: To review studies that evaluated the IES's psychometric properties. METHOD: Literature review. RESULTS: The results indicated that the IES's two-factor structure is stable over different types of events, that it can discriminate between stress reactions at different times after the event, and that it has convergent validity with observer-diagnosed post-traumatic stress disorder. The use of IES in many psychopharmacological trials and outcome studies is supportive of the measure's clinical relevance. CONCLUSIONS: The IES is a useful measure of stress reactions after a range of traumatic events, and it is valuable for detecting individuals who require treatment. PMID- 11872512 TI - Australia's National Mental Health Strategy. AB - BACKGROUND: Australia commenced a 5-year reform of mental health services in 1993. AIMS: To report on the changes to mental health services achieved by 1998. METHOD: Analysis of data from the Australian National Mental Health Report 2000 and an independent evaluation of the National Mental Health Strategy. RESULTS: Mental health expenditure increased 30% in real terms, with an 87% growth in community expenditures, a 38% increase in general hospitals and a 29% decrease in psychiatric hospitals. The growth in private psychiatry, averaging 6% annually prior to 1992, was reversed. Consumer and carer involvement in services increased. CONCLUSIONS: Major structural reform was achieved but there was limited evidence that these changes had been accompanied by improved service quality. The National Mental Health Strategy was renewed for another 5 years. PMID- 11872513 TI - Mental health of teenagers who use cannabis. Results of an Australian survey. AB - BACKGROUND: There is concern in the community about increasing cannabis use and its potential effect on health. AIMS: To ascertain the prevalence of cannabis use among Australian adolescents, associations with mental health problems, risk behaviours and service use. METHOD: Examination of data from a national representative sample of households comprising 1261 adolescents aged 13-17 years. Parents completed a psychiatric interview and questionnaires while adolescents completed questionnaires. RESULTS: One-quarter of the adolescents in the sample had used cannabis. There were no gender differences. Use increased rapidly with age, was more common in adolescents living with a sole parent and was associated with increased depression, conduct problems and health risk behaviours (smoking, drinking) but not with higher use of services. CONCLUSIONS: Cannabis use is very prevalent. The association with depression, conduct problems, excessive drinking and use of other drugs shows a malignant pattern of comorbidity that may lead to negative outcomes. PMID- 11872514 TI - Age, gender and ethnicity of those detained under Part II of the Mental Health Act 1983. AB - BACKGROUND: Aggregate returns give limited information about those detained under the Mental Health Act 1983. AIMS: To use existing data-sets to examine detentions under Part II of the Act. METHOD: Data from 26 areas, with a combined population of 9.2 million, were combined. Population census data were used to standardise rates of detention by age, gender and ethnicity. RESULTS: The 31 702 detentions are distributed bimodally with peaks at age 25-34 years and at over age 80 years. In the younger age group rates of detention are higher for men. The excess of women in the older group is no longer apparent when rates are standardised for age and gender. Detentions are over six times more likely to be of Black people than of White (450 v. 68 per standardised 100 000 population). CONCLUSIONS: The difference in rates of detention between Black and White people is greater than previously thought. The excess of older women detained under Part II of the Act is largely due to the lower life expectancy of men. PMID- 11872515 TI - Annual cost of bipolar disorder to UK society. AB - BACKGROUND: The socio-economic impact of bipolar disorder in the UK is unknown. AIMS: To estimate the annual socio-economic burden imposed by bipolar disorder on UK society. METHOD: The annual cost of resource use attributable to managing bipolar disorder was calculated. Indirect societal costs were also calculated. RESULTS: The annual National Health Service (NHS) cost of managing bipolar disorder was estimated to be 199 million pounds sterling , of which hospital admissions accounted for 35%. The annual direct non-health-care cost was estimated to be 86 million pounds sterling annually and the indirect societal cost was estimated to be 1770 million pounds sterling annually. CONCLUSIONS: The annual cost to UK society attributable to bipolar disorder was estimated to be 2 billion pounds sterling at 1999/2000 prices (estimated 297 000 people with the disorder). Ten per cent of this cost is attributable to NHS resource use, 4% to non-health-care resource use and 86% to indirect costs. PMID- 11872516 TI - Psychotherapy for sexually abused girls: psychopathological outcome findings and patterns of change. AB - BACKGROUND: Controversy exists about the efficacy of psychotherapy for the mental health problems of sexually abused children. AIMS: To compare the relative efficacy of focused individual or group therapy in symptomatic sexually abused girls, and to monitor psychiatric symptoms for persistence or change. METHOD: A multi-centre psychotherapy outcome study recruited 71 sexually abused girls aged 6-14 years who were randomly assigned to focused individual psychotherapy (up to 30 sessions) or psychoeducational group therapy (up to 18 sessions). Changes over the course of the study were monitored. RESULTS: Both treatment groups showed a substantial reduction in psychopathological symptoms and an improvement in functioning, but with no evident difference between individual and group therapy. However, individual therapy led to a greater improvement in manifestations of post-traumatic stress disorder (PTSD). CONCLUSIONS: The beneficial effects on PTSD support the use of individual therapy. However, the small sample size and lack of a control group limit conclusions about changes attributable to treatment. PMID- 11872517 TI - Frequent attenders with medically unexplained symptoms: service use and costs in secondary care. AB - BACKGROUND: Frequent attenders in medical settings account for a disproportionate amount of health-care resources. Little is known about service use and costs of secondary care in those frequent attenders presenting with medically unexplained symptoms. AIMS: To compare health-care use and costs of patients with medically unexplained symptoms with other frequent attenders in secondary care. METHOD: In a sample of 400 frequent attenders of secondary care services, those presenting with medically unexplained symptoms were identified by a review of medical records. Their use of health-care resources was compared with that of other frequent attenders. RESULTS: Of the frequent attenders 17% had at least two medically unexplained consultation episodes. These patients had a greater number of referrals to secondary care and were more likely to undergo particular investigations. CONCLUSIONS: Frequent attenders with medically unexplained symptoms account for levels of service use and expenditure that are comparable with other frequent attenders, but the use and cost of medical investigations in this group are significantly greater. PMID- 11872518 TI - Monitoring community psychiatric services in Italy: differences between patients who leave care and those who stay in treatment. AB - BACKGROUND: Continuity of care has been monitored rarely in Italian community mental health centres. AIMS: To estimate the long-term probability of leaving care in first-contact patients attending an out-patient service, and to identify patients most likely to drop out. METHOD: All patients who had a first contact with the community mental health centre of Magenta during a 1-year recruitment period were followed up for 24 months. Patients who failed to return after the last out-patient visit were regarded as 'drop-outs'. RESULTS: During the 1-year recruitment period 330 subjects were at their first contact. The 1-year incidence of first-contact patients was nearly 33 per 10 000 inhabitants. At follow-up, 46% of patients had dropped out. In comparison with patients with psychoses, subjects suffering from neurotic (P =0.004) and personality disorders (P=0.029) were more likely to drop out. CONCLUSIONS: In the Italian system of community psychiatric care nearly half of the patients are no longer in contact after 2 years. Those who stay in treatment are more likely to suffer from psychosis, suggesting a commitment of Italian out-patient facilities to tackling the needs of patients with more severe disorders. PMID- 11872520 TI - Use of a routine, self-report outcome measure (HoNOSCA-SR) in two adolescent mental health services. Health of the Nation Outcome Scale for Children and Adolescents. AB - BACKGROUND: The Health of the Nation Outcome Scale for Children and Adolescents (HoNOSCA) is an established outcome measure for child and adolescent mental health. Little is known of adolescent views on outcome. AIMS: To develop and test the properties of an adolescent, self-rated version of the scale (HoNOSCA-SR) against the established clinician-rated version. METHOD: A comparison was made of 6-weekly clinician-rated and self-rated assessments of adolescents attending two services, using HoNOSCA and other mental health measures. RESULTS: Adolescents found HoNOSCA-SR acceptable and easy to rate. They rated fewer difficulties than the clinicians and these difficulties were felt to improve less during treatment, although this varied with diagnosis and length of treatment. Although HoNOSCA-SR showed satisfactory reliability and validity, agreement between clinicians and users in individual cases was poor. CONCLUSIONS: Routine outcome measurement can include adolescent self-rating with modest additional resources. The discrepancy between staff and adolescent views requires further evaluation. PMID- 11872519 TI - Observer effects and heritability of childhood attention-deficit hyperactivity disorder symptoms. AB - BACKGROUND: Twin studies have found that childhood attention-deficit hyperactivity disorder (ADHD) has a strong genetic component. Estimates of heritability, the extent of non-additive genetic effects and of 'sibling contrast' effects vary between different studies. AIMS: To use multiple informants to assess the extent to which observer effects influence such estimates in an epidemiological sample of twins. METHOD: Questionnaire packs were sent to the families and teachers of twins aged 5-16 years in the Bro Taf region of South Wales. The twins were ascertained from community paediatric registers. RESULTS: Both parent- and teacher- rated data showed a high degree of heritability for ADHD measured as a symptom dimension, but the correlation between the two types of rater was modest. Bivariate analyses suggested that parent and teacher ratings reflect the effects of different genes. Self-report data from twins aged 11-16 years showed no evidence of genetic effects. CONCLUSIONS: Although ADHD is shown to be highly heritable by both parent- and teacher-rated data, the underlying genotypes may be substantially different. This has implications for study designs aiming to find genes that contribute to the disorder. PMID- 11872521 TI - Islington study of dementia subtypes in the community. AB - BACKGROUND: Epidemiological studies of dementia subtypes have revealed widely varying distribution rates. There are almost no published community prevalence data for dementia with Lewy bodies (DLB) or the frontal lobe dementias (FLD). AIMS: To identify the distribution of dementia subtypes in a representative community population of older people. METHOD: People aged > or = 65 years in randomised enumeration districts in Islington, north London, were screened using a reliable and valid questionnaire. People screened as having dementia were assessed in detail and diagnoses were made according to standard diagnostic criteria. RESULTS: Of 1085 people interviewed, 107 (9.86%) met screening criteria for dementia. Diagnoses were made for 72 people (67.3%). Distribution of subtypes varied according to the criteria used; the best-validated criteria yielding: Alzheimer's disease 31.3%; vascular dementia 21.9%; DLB 10.9%; and FLD 7.8%. CONCLUSIONS: Alzheimer's disease is confirmed as the most common cause of dementia in older people, followed by vascular dementia. However, DLB and FLD occur sufficiently often to be seen frequently in clinical practice and should be incorporated into future editions of standard diagnostic criteria. PMID- 11872522 TI - Genetic risk factors and variation in European suicide rates. PMID- 11872523 TI - Genetic risk factors and variation in European suicide rates. PMID- 11872524 TI - Aggression in schizophrenia: assessment and prevalence. PMID- 11872525 TI - Psychiatric morbidity and elderly offenders. PMID- 11872526 TI - Psychiatric morbidity and elderly offenders. PMID- 11872527 TI - Post-traumatic stress disorder and management of stillbirth. PMID- 11872528 TI - The evolutionary psychology debate. PMID- 11872532 TI - The misdiagnosis of epilepsy. PMID- 11872533 TI - Surgery for temporal lobe epilepsy. PMID- 11872534 TI - The discomfort of patient power. PMID- 11872535 TI - Randomised controlled trials for homoeopathy. PMID- 11872536 TI - Brain drain and health professionals. PMID- 11872537 TI - GPs asked to do more for drug misusers. PMID- 11872538 TI - NHS settles claim of patients treated with LSD. PMID- 11872540 TI - Blair warns of tax increases to pay for NHS. PMID- 11872541 TI - US encourages flu jabs for infants aged under 2. PMID- 11872542 TI - Embryo cell research should continue, committee says. PMID- 11872543 TI - Watchdog approves embryo selection to treat 3 year old child. PMID- 11872544 TI - Bush moves to tighten security around scientific research. PMID- 11872545 TI - US issues new guidelines as mammography debate continues. PMID- 11872546 TI - First cases of type 2 diabetes found in white UK teenagers. PMID- 11872547 TI - Costs and benefits of a one stop clinic compared with a dedicated breast clinic: randomised controlled trial. AB - OBJECTIVE: To determine the cost to the NHS and the impact on anxiety of a one stop clinic for assessing women with suspected breast cancer. STUDY DESIGN: Randomised controlled trial. PARTICIPANTS: Women aged 35 or over referred with a breast lump. STUDY SETTING: Teaching hospital, north west England. INTERVENTIONS: Women were randomly allocated to attend a one stop clinic or a dedicated breast clinic. OUTCOME MEASURES: Reduction in mean anxiety from baseline at 24 hours after the first visit and at 3 weeks and 3 months after diagnosis; mean cost per patient. RESULTS: 670 women were randomised. Compared with women who attended the dedicated clinic, patients attending the one stop clinic were less anxious 24 hours after the visit (adjusted mean change in state anxiety _5.7 (95% confidence interval _8.4 to _3.0)) but not at 3 weeks or 3 months after diagnosis. The additional cost to the NHS of a one stop attendance was pound 32 per woman; this was largely explained by greater cytopathological and radiological staff costs. CONCLUSION: One stop clinics may not be justified in terms of a reduction in short term anxiety. PMID- 11872548 TI - Ethnic differences in invasive management of coronary disease: prospective cohort study of patients undergoing angiography. AB - OBJECTIVES: To compare rates of revascularisation in south Asian and white patients undergoing coronary angiography in relation to the appropriateness of revascularisation and clinical outcome. DESIGN: Prospective cohort study of patients with two and a half years' follow up; appropriateness of revascularisation rated by nine experts with no knowledge of ethnicity of patient. SETTING: Tertiary cardiac centre in London with referral from five contiguous health authorities. PARTICIPANTS: Consecutive patients (502 south Asian, 2974 white) undergoing coronary angiography in the appropriateness of coronary revascularisation study (ACRE). MAIN OUTCOME MEASURES: Coronary revascularisation, non-fatal myocardial infarction, mortality. RESULTS: There was no difference between south Asian and white patients in the proportions deemed appropriate for revascularisation (72% (361) v 68% (2022)) or in the proportions for whom the physician's intended management was revascularisation (39% (196) v 41% (1218)). Among patients appropriate for revascularisation, age adjusted rates of coronary angioplasty (hazard ratio 0.69, 95% confidence interval 0.47 to 1.00, P=0.058) and coronary artery bypass grafting (0.74, 0.58 to 0.91, P=0.007) were lower in south Asian than in white patients. These differences were smaller but still present after adjustment for socioeconomic status and after restriction of analysis to those patients for whom the intended management was revascularisation. There were no differences in mortality and non-fatal myocardial infarction between south Asian and white patients (1.07, 0.78 to 1.47). CONCLUSION: Among patients deemed appropriate for coronary artery bypass grafting, south Asian patients are less likely than white patients to receive it. This difference is not explained by physician bias. PMID- 11872549 TI - Eliciting views of patients with head and neck cancer and carers on professionally derived standards for care. AB - OBJECTIVES: To examine views of patients and carers on the process of care for people with head and neck cancer; to assess whether focus groups are useful in this setting; to compare priorities and standards identified with those published by healthcare professionals; and to incorporate the expressed views into existing national standards. DESIGN: Multicentre study of nine regional focus groups. SETTING: Area covered by two regional health authorities. PARTICIPANTS: 40 patients who had had head and neck cancer and 18 carers. MAIN OUTCOME MEASURES: Views of individuals and groups on standards. Applicability of the method for patients whose appearance and ability to communicate was altered and for recently bereaved carers. Ease of incorporation of views into national and regional standards. RESULTS: Patients and carers participated in discussions on all the principal questions. Opinions were expressed on waiting times, information available to patients, coordination of care, and crisis management. Professionally derived standards were substantially improved by the incorporation of the views of patients and carers. There were no technical problems in carrying out this study on patients with communication difficulties or altered appearance nor with recently bereaved carers. Occasionally, participants said that the meetings were therapeutic. CONCLUSIONS: Professionally facilitated and analysed focus groups are effective in assessing views of patients with cancer and carers on professionally derived standards for care and can be applied in settings traditionally viewed as difficult. Views expressed by patients and carers are powerful motivators for change in the delivery of cancer care. PMID- 11872551 TI - Use of ultramolecular potencies of allergen to treat asthmatic people allergic to house dust mite: double blind randomised controlled clinical trial. AB - OBJECTIVE: To evaluate the efficacy of homoeopathic immunotherapy on lung function and respiratory symptoms in asthmatic people allergic to house dust mite. DESIGN: Double blind randomised controlled trial. SETTING: 38 general practices in Hampshire and Dorset. PARTICIPANTS: 242 people with asthma and positive results to skin prick test for house dust mite; 202 completed clinic based assessments, and 186 completed diary based assessments. INTERVENTION: After a four week baseline assessment, participants were randomised to receive oral homoeopathic immunotherapy or placebo and then assessed over 16 weeks with three clinic visits and diary assessments every other week. OUTCOME MEASURE: Clinic based assessments: forced expiratory volume in one second (FEV(1)), quality of life, and mood. DIARY BASED ASSESSMENTS: morning and evening peak expiratory flow, visual analogue scale of severity of asthma, quality of life, and daily mood. RESULTS: There was no difference in most outcomes between placebo and homoeopathic immunotherapy. There was a different pattern of change over the trial for three of the diary assessments: morning peak expiratory flow (P=0.025), visual analogue scale (P=0.017), and mood (P=0.035). At week three there was significant deterioration for visual analogue scale (P=0.047) and mood (P=0.013) in the homoeopathic immunotherapy group compared with the placebo group. Any improvement in participants' asthma was independent of belief in complementary medicine. CONCLUSION: Homoeopathic immunotherapy is not effective in the treatment of patients with asthma. The different patterns of change between homoeopathic immunotherapy and placebo over the course of the study are unexplained. PMID- 11872553 TI - A comparative case study of two models of a clinical informaticist service. AB - OBJECTIVES: To describe and evaluate two different models of a clinical informaticist service. DESIGN: A case study approach, using various qualitative methods to illuminate the complexity of the project groups' experiences. SETTING: UK primary health care. INTERVENTIONS: Two informaticist projects to provide evidence based answers to questions arising in clinical practice and thereby support high quality clinical decision making by practitioners. RESULTS: The projects took contrasting and complementary approaches to establishing the service. One was based in an academic department of primary health care. The service was academically highly rigorous, remained true to its original proposal, included a prominent research component, and involved relatively little personal contact with practitioners. This group achieved the aim of providing general information and detailed guidance to others intending to set up a similar service. The other group was based in a service general practice and took a much more pragmatic, flexible, and facilitative approach. They achieved the aim of a credible, acceptable, and sustainable service that engaged local practitioners beyond the innovators and enthusiasts and secured continued funding. CONCLUSION: An informaticist service should be judged on at least two aspects of quality---an academic dimension (the technical quality of the evidence based answers) and a service dimension (the facilitation of questioning behaviour and implementation). This study suggests that, while the former may be best achieved within an academic environment, the latter requires a developmental approach in which pragmatic service considerations are addressed. PMID- 11872557 TI - Bradycardias and atrioventricular conduction block. PMID- 11872555 TI - Renal transplantation. PMID- 11872558 TI - The architecture of diagnostic research. PMID- 11872560 TI - Performance league tables. League tables are unreasonably simple. PMID- 11872561 TI - Effect of patient centredness and positive approach. Airing uncertainty can be positive. PMID- 11872562 TI - Somatisation in primary care. Solitary disclosure allows people to determine their own dose. PMID- 11872563 TI - Deputy editor of Clinical Evidence replies to letter. PMID- 11872564 TI - Quality of Cochrane reviews. Quality of Cochrane reviews is better than that of non-Cochrane reviews. PMID- 11872565 TI - Reye's syndrome revisited. Outdated concept of Reye's syndrome was used. PMID- 11872566 TI - Demand for prostate specific antigen testing in primary care. Screening through back passage as well as back door? PMID- 11872567 TI - Despite author's opinion, radiology guideline was correct. PMID- 11872568 TI - Dishonest doctors should not continue to practise. PMID- 11872569 TI - Clinical medication review by pharmacists would improve care. PMID- 11872570 TI - Dementia is being avoided in NHS and social care. PMID- 11872603 TI - Towards the elucidation of the genetic and brain bases of developmental speech and language disorders. PMID- 11872604 TI - Behavioural analysis of an inherited speech and language disorder: comparison with acquired aphasia. AB - Genetic speech and language disorders provide the opportunity to investigate the biological bases of language and its development. Critical to these investigations are the definition of behavioural phenotypes and an understanding of their interaction with epigenetic factors. Here, we report our investigations of the KE family, half the members of which are affected by a severe disorder of speech and language, which is transmitted as an autosomal-dominant monogenic trait. The cognitive manifestations of this disorder were investigated using a number of linguistic and non-linguistic tests. The aims of these investigations were to establish the existence of a 'core' deficit, or behavioural phenotype, and to explain how such a deficit during development might give rise to the range of other impairments demonstrated by affected family members. The affected family members were compared both with the unaffected members and with a group of adult patients with aphasia resulting from a stroke. The score on a test of repetition of non-words with complex articulation patterns successfully discriminated the affected and unaffected family members. The affected family members and the patients with aphasia had remarkably similar profiles of impairment on the tests administered. Pre-morbidly, however, the patients with aphasia had enjoyed a normal course of cognitive development and language experience. This benefit was reflected on a number of tests in which the patients with aphasia performed significantly better than the affected family members and, in the case of some tests, at normal levels. We suggest that, in the affected family members, the verbal and non-verbal deficits arise from a common impairment in the ability to sequence movement or in procedural learning. Alternatively, the articulation deficit, which itself might give rise to a host of other language deficits, is separate from a more general verbal and non-verbal developmental delay. PMID- 11872605 TI - MRI analysis of an inherited speech and language disorder: structural brain abnormalities. AB - Analyses of brain structure in genetic speech and language disorders provide an opportunity to identify neurobiological phenotypes and further elucidate the neural bases of language and its development. Here we report such investigations in a large family, known as the KE family, half the members of which are affected by a severe disorder of speech and language, which is transmitted as an autosomal dominant monogenic trait. The structural brain abnormalities associated with this disorder were investigated using two morphometric methods of MRI analysis. A voxel-based morphometric method was used to compare the amounts of grey matter in the brains of three groups of subjects: the affected members of the KE family, the unaffected members and a group of age-matched controls. This method revealed a number of mainly motor- and speech-related brain regions in which the affected family members had significantly different amounts of grey matter compared with the unaffected and control groups, who did not differ from each other. Several of these regions were abnormal bilaterally, including the caudate nucleus, which was of particular interest because this structure was also found to show functional abnormality in a related PET study. We performed a more detailed volumetric analysis of this structure. The results confirmed that the volume of this nucleus was reduced bilaterally in the affected family members compared with both the unaffected members and the group of age-matched controls. This reduction in volume was most evident in the superior portion of the nucleus. The volume of the caudate nucleus was significantly correlated with the performance of affected family members on a test of oral praxis, a test of non-word repetition and the coding subtest of the Wechsler Intelligence Scale. These results thus provide further evidence of a relationship between the abnormal development of this nucleus and the impairments in oromotor control and articulation reported in the KE family. PMID- 11872606 TI - Changes in visual cortex excitability in blind subjects as demonstrated by transcranial magnetic stimulation. AB - Any attempt to restore visual functions in blind subjects with pregeniculate lesions provokes the question of the extent to which deafferented visual cortex is still able to generate conscious visual experience. As a simple approach to assessing activation of the visual cortex, subjects can be asked to report conscious subjective light sensations (phosphenes) elicited by focal transcranial magnetic stimulation (TMS) over the occiput. We hypothesized that such induction of phosphenes can be used as an indicator of residual function of the visual cortex and studied 35 registered blind subjects after partial or complete long term (>10 years) deafferentation of the visual cortex due to pregeniculate lesions. TMS was applied over the visual cortex in 10 blind subjects with some residual vision (visual acuity <20/400; Group 1), 15 blind subjects with very poor residual vision (only perception of movement or light; Group 2), 10 blind subjects without any residual vision (Group 3) and 10 healthy controls. A stimulation mapping procedure was performed on a 1 x 1 cm skull surface grid with 130 stimulation points overlying the occipital skull. We analysed the occurrence of phosphenes at each stimulation point with regard to frequency and location of phosphenes in the visual field. Previous experiments have shown that repetitive TMS reliably elicits brief flashes of white or coloured patches of light. Therefore, stimulation was performed with short trains of seven consecutive 15 Hz stimuli applied with an intensity of 1.3 times the motor threshold. Under such conditions, phosphenes occurred in 100% of subjects in Group 1, in 60% of Group 2 and in 20% of Group 3. Phosphene thresholds were normal, but the number of effective stimulation sites was significantly reduced in Groups 2 and 3. The results indicate that in blind subjects there is alteration in TMS-induced activation of the deafferented visual cortex or processes engaged in bringing the artificial cortex input to consciousness. The ability to elicit phosphenes is reduced in subjects with a high degree of visual deafferentation, especially in those without previous visual experience. PMID- 11872607 TI - Defective cortical drive to muscle in Parkinson's disease and its improvement with levodopa. AB - We recorded whole-scalp magnetoencephalographic (MEG) signals simultaneously with surface electromyographic (EMG) activity from eight patients with Parkinson's disease after withdrawal and reinstatement of treatment with levodopa. Variations were seen in the coherence between the forearm extensor EMG and the MEG signal originating near or in the hand region of the primary motor cortex. As a group, the parkinsonian patients withdrawn from levodopa showed a reduction in the coherence at 15-30 Hz and 35-60 Hz, and a further three untreated patients had abnormally strong MEG-EMG coherence at 5-12 Hz compared with when medicated or with eight healthy age-matched control subjects. We conclude that the basal ganglia have a specific effect on the temporal organization of motor cortical activity during voluntary tonic contraction. Abnormalities in this aspect of basal ganglia function may directly contribute to bradykinesia and weakness in Parkinson's disease. PMID- 11872608 TI - Sensory determinants of thermal pain. AB - It is still unclear whether the quality of painful thermal sensation is determined only by conduction in specific, dedicated nociceptive channels (i.e. C or Adelta nociceptors) or whether it is a result of integrated activity in both nociceptive and non-nociceptive systems. To evaluate this question, we conducted quantitative and qualitative somatosensory testing in spinal cord injury subjects who suffered from partial or complete loss of thermal sensibility. Testing was performed in skin areas, below the level of the lesion, which were either lacking any thermal sensibility, lacking only one thermal sensation (either heat or cold) or having normal thermal sensations. We found that, in areas lacking any thermal sensibility, warm and cold stimuli produced a sensation of pricking pain, which had no thermal quality and was detected at significantly higher thresholds than in normal controls (48.5 +/- 1.8 and 9.7 +/- 5.1 degrees C for noxious heat- and noxious cold-induced pricking pain, respectively). Normal thermal pain sensations, consisting of normal perception of thermal quality and normal mean pain thresholds, were present both in normal skin areas (42.1 +/- 1.9 and 27.6 +/ 2.25 degrees C for heat and cold pain, respectively) and in areas in which only one thermal modality remained intact, when tested for that modality. Thus, testing for heat pain in areas in which only warm sensation was intact, or cold pain when only cold was intact produced normal qualities and thresholds of pain (42.8 +/- 3.4 and 24.4 +/- 6.2 degrees C for heat and cold pain, respectively). No spatial summation of pricking pain was observed, in contrast to the marked summation of heat pain in normal areas. In areas with only a single intact thermal modality, the quality of the perceived non-painful sensation was not determined by the thermal stimulus but by the intact modality (paradoxical sensation). Cold stimuli were perceived as warm in areas in which only warm sensation was preserved, and vice versa. A similar pattern was also seen for pain perception in areas with intact warm sensation. In these areas, both noxious heat and cold elicited a sensation of heat pain. No consistent pattern of heat elicited pain was observed in areas in which only cold sensation was intact. These data suggest that the integrity of non-noxious thermal systems is essential for the normal perception of thermal pain, and that the subjective sensation of pain depends on the integration of information from nociceptive and non nociceptive channels. PMID- 11872609 TI - Deficits of musical timbre perception after unilateral temporal-lobe lesion revealed with multidimensional scaling. AB - Thirty patients with unilateral temporal lobe excisions and 15 normal control subjects were tested in a task involving judgements of timbre dissimilarity in single tone and melodic conditions. Perceptual correlates of spectral and temporal parameters resulting from changing the number of harmonics and rise-time duration, respectively, were investigated by using a multidimensional scaling technique. The results of subjects with left temporal lobe lesion suggest that they were able to use the spectral and temporal envelopes of tones independently in making perceptual judgements of single tones. In the melodic condition, their results were significantly different from those of normal control subjects, suggesting that left temporal lesions do affect subtle aspects of timbre perception, despite these patients' preserved ability to make discrimination judgements using traditional paradigms. The major finding of this study concerns perceptual ratings obtained by subjects with right temporal lobe lesion, which revealed a disturbed perceptual space in both conditions. The most distorted results were obtained with single tones, in which the temporal parameter was less prominent. Tones were grouped according to their spectral content, but the results did not reflect a coherent underlying perceptual dimension. In general, the data from both patient groups (left lesions and right lesions) showed that the extraction of temporal cues was easier in the melodic than in the single tone condition, suggesting that the different durations and frequencies heard in a musical phrase enhance the importance of certain physical parameters. The findings of the present study replicate and extend previous results showing that timbre perception depends mainly upon the integrity of right neocortical structures, although a contribution of left temporal regions is also apparent. These data also demonstrate that multidimensional techniques are sensitive to more subtle perceptual disturbances that may not be revealed by discrimination paradigms. PMID- 11872610 TI - Local improvement in auditory frequency discrimination is associated with hearing loss slope in subjects with cochlear damage. AB - Earlier data in the literature have shown local improvements in frequency discrimination performance near the cut-off frequency of steeply sloping, high frequency hearing loss in subjects with cochlear damage. The general objective of the present study was to characterize further the relationships between this effect and various audiometric variables: namely, the slope, extent and shape of the hearing loss. In particular, we were interested in determining whether the effect was present in subjects with more moderately sloping hearing loss and/or other patterns of loss. Frequency difference limens (DLFs) were measured in 20 subjects (eight female, 12 male, median age 55.5 years) with high-frequency hearing loss. At least 12 frequencies were tested at intervals of 1/8 octave over a range of 1.5 octaves around the cut-off frequency for hearing loss (Fc). The Fc corresponded to the audiogram edge frequency and was defined as the highest test frequency, at the beginning of the slope, with a hearing threshold of no more than 5 dB HL above that of the best hearing frequency. The level of the test tones was randomized over a range of 6 dB around a nominal level, following an equal-loudness contour curve measured at 1/2-octave intervals. Results showed that DLFs were significantly smaller in a frequency band 1/4 octave wide centred on Fc than in the other bands. Furthermore, the average DLF measured in this band proved to be negatively correlated with the slope of hearing loss. No such significant relationship was found with the other audiometric indices considered, namely, the extent and maximum amount of hearing loss and the log-transformed cut off frequency. The 20 subjects were divided into three groups according to the slope of their hearing loss relative to Fc (steep, >25 dB/1/2 octave; medium, between 12 and 25 dB/1/2 octave; and shallow, <12 dB/1/2 octave). A local improvement in DLF around Fc was observed in the steep- and medium-slope groups and was confirmed statistically in the steep-slope group. Similar measurements in subjects with low-frequency or notched hearing loss allowed us to establish the presence of similar local improvements in DLFs around audiogram edges. These results, which suggest the slope of the hearing loss to be the most important factor for the occurrence of local DLF improvements, are consistent with both an interpretation in terms of peripheral mechanisms and one in terms of central mechanisms, i.e. injury-induced neural reorganization. PMID- 11872611 TI - Oligoclonal expansion of memory CD8+ T cells in cerebrospinal fluid from multiple sclerosis patients. AB - Multiple sclerosis is a chronic inflammatory demyelinating disease of the CNS. Although the aetiology of multiple sclerosis is still unknown, it is widely believed that T cells play a central role in its pathogenesis. To identify and characterize disease-relevant T cells, we analysed CD4+ and CD8+ T cells freshly isolated from the CSF and peripheral blood of 36 multiple sclerosis patients for their T-cell receptor variable beta (TCRBV) chain repertoire. In most patients, we found significant overexpression of individual TCRBV chains on CD8+ T cells from CSF compared with peripheral blood. In contrast, only a few multiple sclerosis patients showed differences between the two compartments in TCRBV expression on CD4+ T cells. The overexpression of specific TCRBV chains on CD8+ T cells was found to be stable over several months in selected patients and involved mainly T cells with a memory phenotype. In two patients studied, individual TCRBV chain overexpression was found to be caused by the expansion of T cell populations with identical or highly similar rearranged T-cell receptor beta- and alpha-chain sequences, which were not found among peripheral blood CD8+ T cells. Our findings demonstrate selective enrichment of memory CD8+ T cells in the CSF of multiple sclerosis patients, suggesting a role for these CD8+ T cells in the pathogenesis of multiple sclerosis. Our study provides a basis for future trials to identify disease-associated antigens and disease pathogenesis in multiple sclerosis. PMID- 11872612 TI - Patients lacking the major CNS myelin protein, proteolipid protein 1, develop length-dependent axonal degeneration in the absence of demyelination and inflammation. AB - Axonal degeneration contributes to clinical disability in the acquired demyelinating disease multiple sclerosis. Axonal degeneration occurs during acute attacks, associated with inflammation, and during the chronic progressive phase of the disease in which inflammation is not prominent. To explore the importance of interactions between oligodendrocytes and axons in the CNS, we analysed the brains of rodents and humans with a null mutation in the gene encoding the major CNS myelin protein, proteolipid protein (PLP1, previously PLP). Histological analyses of the CNS of Plp1 null mice and of autopsy material from patients with null PLP1 mutations were performed to evaluate axonal and myelin integrity. In vivo proton magnetic resonance spectroscopy (MRS) of PLP1 null patients was conducted to measure levels of N-acetyl aspartate (NAA), a marker of axonal integrity. Length-dependent axonal degeneration without demyelination was identified in the CNS of Plp1 null mice. Proton MRS of PLP1-deficient patients showed reduced NAA levels, consistent with axonal loss. Analysis of patients' brain tissue also demonstrated a length-dependent pattern of axonal loss without significant demyelination. Therefore, axonal degeneration occurs in humans as well as mice lacking the major myelin protein PLP1. This degeneration is length dependent, similar to that found in the PNS of patients with the inherited demyelinating neuropathy, CMT1A, but is not associated with significant demyelination. Disruption of PLP1-mediated axonal--glial interactions thus probably causes this axonal degeneration. A similar mechanism may be responsible for axonal degeneration and clinical disability that occur in patients with multiple sclerosis. PMID- 11872613 TI - Global attentional-executive sequelae following surgical lesions to globus pallidus interna. AB - It has been demonstrated that selective unilateral surgical ablation of posteroventral globus pallidus interna relieves the movement disorders associated with advanced Parkinson's disease, without necessarily incurring the executive cognitive sequelae that have been observed following gross pathological lesions to this brain region. This finding is consistent with established theory that underlying neuronal circuitry is functionally segregated into parallel cortico striatal-pallidal-thalamo-cortical 'loops'. We have studied a series of 12 patients with advanced Parkinson's disease at baseline, and then following bilateral pallidotomy, with a battery of neuropsychological tests including the Cambridge Neuro psychological Test Automated Battery. We identified a selective and universal loss of individual patients' ability to shift attention to novel dimensions in a test of abstract rule-learning following surgery, which was not reliably associated with any other change in cognition, personality, mood or medication. This finding is rare in its specificity and has implications for theoretical models of the functional architecture and pathophysiology of the globus pallidus, and the clinical practice of pallidotomy. PMID- 11872614 TI - Clinical predictive factors of subthalamic stimulation in Parkinson's disease. AB - High-frequency stimulation of the subthalamic nucleus (STN) constitutes one of the most effective treatments for advanced forms of Parkinson's disease. The cost and potential risks of this procedure encourage the determination of clinical characteristics of patients that will have the best postoperative outcome. Forty one Parkinson's disease patients underwent surgery for bilateral STN stimulation. The selection criteria were severe parkinsonian motor disability, clear response of symptoms to levodopa, occurrence of disabling levodopa-related motor complications and the absence of dementia and significant abnormalities on brain MRI. Clinical evaluation was performed 1 month before and 6 months after surgery. The improvement in the activities of daily living subscale of the Unified Parkinson's Disease Rating Scale, Part II (UPDRS II) and parkinsonian motor disability (UPDRS III) was greater when the preoperative scores for activities of daily living and parkinsonian motor disability, in particular axial symptoms, such as gait disorders and postural instability assessed at the time of maximal clinical improvement (on drug), were lower. Age and disease duration were not predictive, but parkinsonian motor disability tended to be more improved in patients with younger age and shorter disease duration. The severity of levodopa related motor complications was not a predictive factor. The outcome of STN stimulation was excellent in levodopa-responsive forms of Parkinson's disease, i.e. in patients with selective brain dopaminergic lesions, and moderate in patients with axial motor symptoms and cognitive impairment known to be less responsive or unresponsive to levodopa treatment, i.e. when brain non dopaminergic lesions develop in addition to the degeneration of the nigrostriatal dopaminergic system. The results are consistent with the classical inclusion criteria for STN stimulation, but imply that the decision to operate on the oldest patients and/or patients with gait and postural disorders, who are poorly responsive to levodopa, should be weighed carefully. PMID- 11872615 TI - Dopaminergic modulation of high-level cognition in Parkinson's disease: the role of the prefrontal cortex revealed by PET. AB - This study examined the effects of L-dopa medication in patients with Parkinson's disease on cortical and subcortical blood flow changes during two tasks known to involve frontostriatal circuitry. Eleven patients with Parkinson's disease were scanned on two occasions, one ON L-dopa medication and one OFF L-dopa medication, during performance of the Tower of London planning task and a related test that emphasized aspects of spatial working memory. L-dopa-induced decreases were observed in the right dorsolateral prefrontal cortex during performance of both the planning and the spatial working memory tasks compared with the visuomotor control task. Conversely, L-dopa-induced blood flow increases were observed in the right occipital lobe during the memory task relative to the control task. Data from age-matched healthy volunteers demonstrated that L-dopa effectively normalized blood flow in these regions in the patient group. Moreover, a significant correlation was found between L-dopa-induced, planning related blood flow decreases in the right dorsolateral prefrontal cortex and L-dopa-induced changes in performance on the planning task. These data suggest that L-dopa ameliorates high-level cognitive deficits in Parkinson's disease by inducing relative blood flow changes in the right dorsolateral prefrontal cortex. PMID- 11872616 TI - Variability of cerebral blood volume and oxygen extraction: stages of cerebral haemodynamic impairment revisited. AB - The presence or degree of haemodynamic impairment due to occlusive cerebrovascular disease is often inferred from measurements of cerebral blood flow (CBF), cerebral blood volume (CBV), oxygen extraction fraction (OEF) and the cerebral rate for oxygen metabolism (CMRO2). However, the relationship of these variables, in particular CBV, to regional cerebral haemodynamics is not clearly established in humans with subacute or chronic disease. In the present study, we investigated the relationship of CBV to OEF, CBF and CMRO2, and to subsequent stroke risk in patients with unilateral carotid artery occlusion, in order to define better the associated haemodynamic and metabolic changes. We reviewed data from 81 patients with symptomatic carotid occlusion enrolled in a prospective study of haemodynamic factors and stroke risk. Measurements of CBV, CBF, OEF and CMRO2 were made on entry using PET. Patients were divided into groups by hemispheric ratios and absolute ipsilateral values of OEF and CBV, based on comparison with normal controls. Haemodynamic and metabolic values, risk factors and stroke risk were compared between groups. Based on hemispheric ratios, 45 patients had increased ipsilateral OEF; CBV was increased in 19 of these 45 patients. No differences in CBF, CMRO2 or clinical risk factors were found between these 19 patients and the remaining 26 patients with increased OEF and normal or reduced CBV. Thirteen ipsilateral strokes occurred during follow-up, and 10 of the 13 occurred in the 19 patients with increased OEF and CBV (log rank P < 0.0001). Thirty-two of the 68 patients with complete quantitative PET data had increased OEF by absolute ipsilateral values. CBV was increased in 20 of the 32 patients. No differences in CBF, CMRO2 or clinical risk factors were found between these 20 patients and the remaining 12 patients with increased OEF and normal CBV. Seven of the nine ipsilateral strokes that occurred in the 68 patients occurred in those 20 patients with increased OEF and increased CBV (log rank P = 0.003). The higher risk of ischaemic stroke in patients with increased OEF and CBV suggests that their degree of haemodynamic compromise is more severe than those with increased OEF and normal CBV. In patients with chronic carotid occlusion and increased OEF, increased CBV may indicate pronounced vasodilation due to exhausted autoregulatory vasodilation. The physiological explanation for the measurement of normal CBV in patients with increased OEF is less certain and may reflect preserved autoregulatory capacity. PMID- 11872617 TI - Long-lasting amelioration of visuospatial neglect by prism adaptation. AB - It has been shown that unilateral left neglect can be significantly improved for a short time after a short period of adaptation to a prismatic shift of the visual field to the right. In neuropsychological studies, however, there is no evidence demonstrating long-lasting effects following treatment by prism adaptation (PA). The first aim of the present study was to find out whether the short-term amelioration found after prismatic adaptation could be converted into long-term therapeutic improvement. Secondly, we investigated whether the improvement of neglect in standard tests could be generalized to ecological visuospatial tests. Thirdly, the effects of prism adaptation on different spatial domains (far, near and personal space) were evaluated. Fourthly, the influence of PA on high-order visuospatial functions, such as spatial representation, and on a low-order factor, i.e. sensory--motor bias, was investigated. Finally, we investigated the possible correlation between neglect amelioration, the adaptation effect and the visuomotor after-effect, as assessed by a pointing task during and after PA. Seven patients with right hemisphere lesion and left visuospatial neglect were treated with prismatic lenses in twice-daily sessions over a period of 2 weeks. In each training session, patients were required to perform a pointing task wearing base-left wedge prisms inducing a shift of the visual field to the right by 10. The presence of visual neglect and the duration of the amelioration achieved were assessed before the treatment and 2 days, 1 week and 5 weeks after treatment by using a standardized battery that included a series of behavioural and ecological visuospatial tests. Six control, untreated patients, matched to the experimental group for gravity and duration of illness, were submitted to the same tests at the same intervals as the experimental patients. The results showed an improvement in the experimental patients' performance after PA, which was maintained during the 5-week period after treatment. The amelioration of neglect was found in standard as well as in behavioural tests and in all spatial domains. In contrast, control patients did not show any improvement in neglect. The amelioration of neglect occurred only in patients who showed the adaptation effect and the after-effect in the pointing task. Neglect amelioration did not occur in one patient who did not show the adaptation effect and had an unstable after-effect. In conclusion, these findings show that prism adaptation is a productive way of achieving long-lasting improvements in neglect treatment. PMID- 11872618 TI - Decision-making processes following damage to the prefrontal cortex. AB - Recent work has suggested an association between the orbitofrontal cortex in humans and practical decision making. The aim of this study was to investigate the profile of cognitive deficits, with particular emphasis on decision-making processes, following damage to different sectors of the human prefrontal cortex. Patients with discrete orbitofrontal (OBF) lesions, dorsolateral (DL) lesions, dorsomedial (DM) lesions and large frontal lesions (Large) were compared with matched controls on three different decision-making tasks: the Iowa Gambling Task and two recently developed tasks that attempt to fractionate some of the cognitive components of the Iowa task. A comprehensive battery including the assessment of recognition memory, working memory, planning ability and attentional set-shifting was also administered. Whilst combined frontal patients were impaired on several of the tasks employed, distinct profiles emerged for each patient group. In contrast to previous data, patients with focal OBF lesions performed at control levels on the three decision-making tasks (and the executive tasks), but showed some evidence of prolonged deliberation. DL patients showed pronounced impairment on working memory, planning, attentional shifting and the Iowa Gambling Task. DM patients were impaired at the Iowa Gambling Task and also at planning. The Large group displayed diffuse impairment, but were the only group to exhibit risky decision making. Methodological differences from previous studies of OBF patient groups are discussed, with particular attention to lesion laterality, lesion size and psychiatric presentation. Ventral and dorsal aspects of prefrontal cortex must interact in the maintenance of rational and 'non-risky' decision making. PMID- 11872619 TI - Seizure anticipation in human neocortical partial epilepsy. AB - The transition of brain activity towards an epileptic seizure is still a poorly understood phenomenon. Dynamic changes in brain activity have been detected several minutes before seizure emergence in populations of patients with mesial temporal lobe epilepsy (MTLE), using non-linear analysis of intracranial EEG recordings. Similar detection of a pre-ictal state has been obtained with standard scalp EEG recordings using a modified non-linear method. Here we applied this strategy to the seizures of patients with neocortical partial epilepsy. Results obtained by non-linear similarity analysis of 41 seizures from 11 patients with refractory partial epilepsy originating from various sites of the neocortex showed that (i) a pre-ictal state was detected in 90% of the patients and in 83% of the seizures whatever their location, with a mean anticipation time of 7.5 min; (ii) similar pre-ictal dynamic changes were detected when non-linear analysis methods were applied to either intracranial or scalp EEG recordings; (iii) the recording sites exhibiting these pre-ictal changes were distributed both within the epileptogenic focus and at remote locations; (iv) most pre-ictal dynamic changes were not correlated with linear changes in the frequency spectrum or with changes in the visually inspected EEG and the patients' behaviour. Hypotheses on the neuronal mechanisms underlying the pre-ictal period are discussed. The present results, together with those recently obtained in an MTLE population, suggest that changes in pre-ictal dynamics are a general phenomenon associated with seizure emergence in a wide population of patients with partial epilepsy, wherever the epileptogenic focus is located. The possibility of anticipating the onset of seizures has considerable therapeutic implications. PMID- 11872620 TI - Neuronal intranuclear inclusions in SCA2: a genetic, morphological and immunohistochemical study of two cases. AB - Spinocerebellar ataxia 2 (SCA2) belongs to the family of autosomal dominant cerebellar ataxias (ADCA), a genetically heterogeneous group of neurodegenerative diseases. The SCA2 gene maps to chromosome 12q24 and the causative mutation involves the expansion of a CAG repeat within the coding region of the gene. Pathologically, SCA2 presents as olivo-ponto-cerebellar atrophy (OPCA). We present the cases of a 41-year-old man and a 54-year-old woman who died after a long illness characterized by severe cerebellar ataxia. Diagnosis of SCA2 was confirmed by genetic analysis. The brains were moderately to severely atrophic and atrophy was particularly obvious in the cerebellum and brainstem. Histological examination revealed extreme loss of pontine and olivary nuclei and Purkinje cells, with preservation of the dentate nuclei, and of the pigmented cells in the substantia nigra. The whole spinal cord was also severely affected, with shrinkage of the dorsal columns and reduction in the number of neurones in the motor pool and Clarke's nuclei. Immunohistochemistry with 1C2 antibody showed granular neuronal cytoplasmic deposits in all the areas examined and widespread intranuclear inclusions, which were particularly numerous in the residual pontine nuclei. Intranuclear inclusions were not considered a feature in SCA2. Our results support the view that intranuclear inclusions are an integral part of the pathology of this mutation. PMID- 11872621 TI - Evidence for axonal membrane hyperpolarization in multifocal motor neuropathy with conduction block. AB - Multiple nerve excitability measurements were used to investigate axonal membrane properties of patients diagnosed with multifocal motor neuropathy (MMN). Six patients were selected, all with evidence of distal focal motor conduction block involving the median nerve in the forearm. In all patients, the median nerve was stimulated at the wrist, just distal to the site of block, and the resulting compound muscle action potentials were recorded from abductor pollicis brevis. Stimulus-response behaviour, the strength--duration time constant, threshold electrotonus to 100 ms polarizing currents, a current-threshold relationship and the recovery of excitability following supramaximal activation were recorded using a protocol described recently. When compared with control values, patients demonstrated significantly greater superexcitability, a 'fanning out' of threshold electrotonus recordings, and a significant change in the slope of the current--threshold relationship. These abnormalities in axonal membrane excitability parameters closely resembled those in normal axons hyperpolarized following release from ischaemia. To test for axonal hyperpolarization, DC depolarizing currents were applied to the nerves of three patients, and all the excitability parameters were normalized by depolarization. Attempts to trace excitability measures proximally towards the site of block were unsuccessful, as the nerve became inexcitable in all cases. It is suggested that the distal hyperpolarization is probably linked to focal depolarization and that the clinical features of MMN are consistent with a depolarizing/hyperpolarizing lesion. PMID- 11872623 TI - Retinal haemorrhages in infant head injury. PMID- 11872622 TI - Neuropathology of inflicted head injury in children. PMID- 11872625 TI - Reporting clinical trials: full access to all the data. PMID- 11872626 TI - Women who smoke: are women more susceptible to tobacco-induced lung cancer? PMID- 11872627 TI - Variable promoter region CpG island methylation of the putative tumor suppressor gene Connexin 26 in breast cancer. AB - Intercellular communication via gap junctions is a mechanism for tumor suppression. Connexin 26 (Cx26) is a structural component of gap junctions expressed by breast epithelial cells. Expression levels of Cx26 are reduced in many breast tumors. Methylation-sensitive single-stranded conformation analysis showed variable methylation in the promoter region CpG island in 11 out of 20 (55%) breast cancer patients. Heterogeneity in methylation patterns was observed both between and within tumors. The degree of methylation ranged from a few CpG dinucleotides to almost all the CpG dinucleotides in the analyzed region. The most frequently methylated CpG was in an Sp1 site known to be important for Cx26 gene expression. One of eight breast cancer cell lines (MD-MBA-453) was methylated in the promoter region and did not express Cx26. Treatment of MDA-MB 453 with 5-aza-2'-deoxycytidine resulted in the re-expression of Cx26 mRNA. Methylation of the promoter region is likely to be an important mechanism in modulating the expression of Cx26 in breast cancer. PMID- 11872628 TI - Overexpression of alpha(v)beta6 integrin in serous epithelial ovarian cancer regulates extracellular matrix degradation via the plasminogen activation cascade. AB - Recent evidence suggests that integrins are involved in the multi-step process of tumour metastasis. The biological relevance of alpha(v) integrins and associated beta-subunits in ovarian cancer metastasis was examined by analysing the expression of these cell surface receptors in nine ovarian cancer cell lines and also in the primary human ovarian surface epithelial cell line (HOSE). beta1, beta3 and beta5 subunits were present in all ten ovarian cell lines. beta6 subunit was present at varying levels in eight out of nine cancer cell lines but was absent in the HOSE cell line. Immunohistochemical staining showed that beta6 was present in both non-invasive (borderline) and high-grade ovarian cancer tissues but was absent in benign and normal ovarian tissue. High alpha(v)beta6 integrin expressing ovarian cancer cell lines had high cell surface expression of uPA and uPAR. Ovarian cancer cell lines expressing high to moderate level of alpha(v)beta6 integrin demonstrated ligand-independent enhanced levels of high molecular weight (HMW)-uPA and pro-matrix metalloproteinase 2 and 9 (pro-MMP-2 and pro-MMP-9) expression in the tumour-conditioned medium. High and moderate expression of alpha(v)beta6 integrin correlated with increased plasminogen dependent degradation of extracellular matrix which could be inhibited by inhibitors of plasmin, uPA and MMPs or by monoclonal antibody against uPA, MMP-9 or alpha(v)beta6 integrin. These results suggest that endogenous de novo expression of alpha(v)beta6 integrin in ovarian cancer cells may contribute to their invasive potential, and that alpha(v)beta6 expression may play a role in ovarian cancer progression and metastasis. PMID- 11872629 TI - Increased expression of cyclooxygenase-2 protein during rat hepatocarcinogenesis caused by a choline-deficient, L-amino acid-defined diet and chemopreventive efficacy of a specific inhibitor, nimesulide. AB - Expression of cyclooxygenase (COX)-2 protein during rat hepatocarcinogenesis associated with fatty change, fibrosis, cirrhosis and oxidative DNA damage, caused by a choline-deficient, L-amino acid-defined (CDAA) diet were investigated in F344 male rats, along with the chemopreventive efficacy of the specific COX-2 inhibitor, nimesulide (NIM). Nimesulide, which was administered in the diet at concentrations of 200, 400, 600 and 800 p.p.m. for 12 weeks, decreased the number and size of preneoplastic enzyme-altered liver foci, levels of oxidative DNA damage, and the grade and incidence of fibrosis in a dose-dependent manner. A preliminary long-term study of 65 weeks also revealed that 800 p.p.m. NIM decreased the multiplicity of neoplastic nodules and hepatocellular carcinomas and prevented the development of cirrhosis. Western blot analysis revealed that COX-2 protein was barely expressed in control livers and increased approximately 2.9-fold in the livers of rats fed on a CDAA diet for 12 weeks and approximately 4.5-5.4-fold in tumors, with a diameter larger than 5 mm, at 80 weeks. Immunohistochemically, COX-2 protein was positive in sinusoidal and stromal cells in fibrotic septa, which were identified by immunoelectron microscopy as Kupffer cells, macrophages, either activated Ito cells or fibroblasts, after exposure to the CDAA diet for 12 weeks, whereas it was only occasionally weakly positive in sinusoidal, probably Kupffer, cells in control livers. In neoplastic nodules in rats fed on a CDAA diet for 30 and 80 weeks, sinusoidal cells and cells with relatively large round nuclei and scanty cytoplasm were strongly positive for COX 2 protein, with the neoplastic hepatocytes in the minority of the nodules, but not the cancer cells, being moderately positive. These results clearly indicate that rat hepatocarcinogenesis, along with fatty change, fibrosis and cirrhosis, is associated with increased expression of COX-2 protein, and point to the chemopreventive efficacy of a selective COX-2 inhibitor against, at least, the early stages of hepatocarcinogenesis. PMID- 11872630 TI - Cyclin D1 gene polymorphism is associated with an increased risk of urinary bladder cancer. AB - Cyclin D1 is believed to play an important role in the genesis and/or progression of transitional cell cancer (TCC) of the urinary bladder. Cyclin D1 gene (CCND1) mRNA is alternatively spliced to produce two transcripts, and the splicing pattern may be modulated by a G to A single nucleotide polymorphism within the splice donor site of exon 4. This study was conducted to explore the association between the polymorphism and the susceptibility to and disease status of TCC of the bladder in 222 cases and 317 native Japanese controls. The relationship between the CCND1 polymorphism and the mRNA splicing pattern in TCC cells was evaluated by semi-quantitative reverse-transcription PCR. The CCND1 A allele was more frequently observed in the TCC group than the control group (P = 0.032) with a significant difference in the genotype frequency between the two groups (P = 0.029). The AA genotype was associated with a significantly higher risk of TCC compared with the AG+GG genotypes (adjusted odds ratio (aOR) = 1.76, 95% confidence interval (CI) = 1.09-2.84, P = 0.022). This association was observed more significantly in nonsmoking cases (aOR = 2.53; 95% CI = 1.28-4.51, P = 0.008). Looking at tumor grade, the presence of the A allele was associated with higher grade (= grade 3) tumors with a gene dosage effect (aOR = 1.77, CI = 1.16 2.69, P = 0.008). In tumor stage, although not significant, the AA + AG genotypes tended to be more frequently observed in cases with T1-4 tumors than those with Ta tumors (aOR = 1.94, 95% CI = 0.98-3.82, P = 0.057). The genotype seemed to influence the two alternatively spliced forms of the CCND1 mRNA because the ratio of the CCND1 transcript-b/transcript-a was significantly higher in cases with the AA genotype compared with those with the AG + GG genotypes. These data suggest that the CCND1 variant A allele may be associated with an increased risk of TCC of the bladder, especially in men without a history of smoking, and it may also have an effect on its disease status. PMID- 11872631 TI - Development of a multi-organ rat model for evaluating chemopreventive agents: efficacy of indole-3-carbinol. AB - Indole-3-carbinol (I-3-C) is among the most widely and popularly known antiestrogens. Due to its putative chemopreventive action, I-3-C is being marketed to the general public in health food establishments. Although it has been demonstrated to prevent cancer in animal bioassays, I-3-C also acts as a promoter in the liver and colon. Because of this potential dual biological activity, it is important to investigate both the inhibitory and promotional activities of I-3-C in multi-organ tumorigenesis animal models. 7,12 Dimethylbenz[a]anthracene, aflatoxin B1 and azoxymethane were used to initiate mammary, liver and colon carcinogenesis, respectively in female Sprague-Dawley rats. The rats were fed continuously on a diet containing I-3-C for 25 weeks after initiation. I-3-C treatment was begun one week after the last carcinogen treatment had been administered. I-3-C treatment resulted in a delay in latency of mammary tumor formation, but did not alter tumor incidence or multiplicity among survivors. In the colon, the protocol produced a 40% decrease in aberrant colon crypt foci. However, in the liver, it strongly-induced GST-P foci in carcinogen-treated (a four-fold increase in volume percent foci) and in the vehicle controls (a 69-fold increase). These data support previous findings in other rodent and fish tumor models that I-3-C both inhibits and promotes carcinogenesis. The results of this study clearly demonstrate that I-3-C is not an appropriate chemoprotective agent for human use, in spite of its effects in the breast and colon in this rat animal model. PMID- 11872632 TI - Analysis of 8-oxo-7,8-dihydro-2'-deoxyguanosine and DNA strand breaks in white blood cells of occupationally exposed workers: comparison with ambient monitoring, urinary metabolites and enzyme polymorphisms. AB - The relationship between biomarkers of effect (8-oxo-7,8-dihydro-2' deoxyguanosine (8-oxodGuo, HPLC system) and tail extent moment (comet assay)), markers of external and internal exposure, and biomarkers of susceptibility was evaluated for coke-oven and graphite-electrode-producing plant workers exposed to polycyclic aromatic hydrocarbons (PAHs). Mean 8-oxodGuo levels in white blood cells (WBC) of exposed workers were between 1.38 times (coke-oven, n = 20; P < 0.01) and 2.15 times (graphite-electrode-producing plant, n = 30; P < 0.01) higher than levels found in control samples (mean +/- SD 0.52 +/- 0.16 8 oxodGuo/10(5) dGuo, n = 47). The mean tail extent moment in lymphocytes was 1.38 times higher for coke-oven workers (n = 19; P = 0.09) and 3.13 times higher for graphite-electrode-producing plant workers (n = 29; P < 0.01) when compared with controls (mean plus minus SD 2.54 +/- 0.68, n = 32). Elevated tail extent moments (>3.73) were found in the majority (84%) of PAH-exposed workers showing increased DNA adduct levels (>0.78 8-oxodGuo/10(5) dGuo). However, no association (P > 0.05) was found between DNA damage (8-oxodGuo/10(5) dGuo or tail extent moment) in WBC of all PAH-exposed workers and either benzo[a]pyrene levels or the sum of 16 PAH levels in the air at work place. Furthermore, no relation (P > 0.05) could be established between DNA damage in WBC and biomarkers of internal exposure (1 hydroxypyrene (1-OHP) and sum of five hydroxyphenanthrenes (OHPHs)). Higher exposure to airborne pyrene and phenanthrene led to increasing concentrations of the metabolites 1-OHP (P < 0.01) and the sum of five OHPHs (P < 0.01) in the urine of PAH-exposed workers. The polymorphisms of genes CYP1A1, GSTM1, GSTT1 and GSTP1 (biomarkers of susceptibility) showed no association with biomarkers of effect. In conclusion, both biomarkers of effect may be appropriate for further surveillance studies of workers under PAH exposure. PMID- 11872633 TI - Chemoprevention of 2-amino-1-methyl-6-phenylimidazo- [4,5-b]pyridine-induced colon carcinogenesis by 1-O-hexyl-2,3,5-trimethylhydroquinone after initiation with 1,2-dimethylhydrazine in F344 rats. AB - The aim of this study is to investigate the chemopreventive effects of the synthetic phenolic antioxidant 1-O-hexyl-2,3, 5-trimethylhydroquinone (HTHQ) on 2 amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP)-associated colon carcinogenesis in rats after initiation with 1,2-dimethylhydrazine (DMH) in male F344 rats. Groups of 20-22, 6-week-old male F344 rats were given four subcutaneous injections of 40 mg/kg body wt of DMH during the initial 4 weeks. They were then maintained on powdered basal diet containing 0.03% PhIP alone, PhIP together with 0.5 or 0.125% HTHQ, 0.5 or 0.125% HTHQ alone or basal diet for 32 weeks. A small number (1.1 +/- 1.1/rat) of colon tumors were induced by DMH treatment alone. After initiation with DMH, the number of colon tumors was greatly increased to 8.3 +/- 5.6 by the administration of PhIP. Additional treatment with HTHQ dose-dependently decreased the multiplicity of colon adenocarcinomas to 4.9 +/- 2.8 and 2.6 +/- 1.4 with 0.125 and 0.5%, respectively. This treatment similarly reduced atypical hyperplasias of the ventral prostate. Furthermore, HTHQ significantly reduced the multiplicity of duodenal adenocarcinomas induced by DMH + PhIP or DMH alone. Immunohistochemically, HTHQ was revealed to suppress PhIP-DNA adduct formation in the epithelial cells of the colon and prostate in a separate 2 weeks experiment. The present results clearly showed that HTHQ has chemopreventive potential for PhIP-associated colon and prostate carcinogenesis. The observed inhibition may largely be due to interference with PhIP-DNA adduct formation. In addition, HTHQ has been demonstrated to inhibit duodenal carcinogenesis in the post-initiation stage. PMID- 11872635 TI - Modulation of repair of ultraviolet damage in the host-cell reactivation assay by polymorphic XPC and XPD/ERCC2 genotypes. AB - DNA repair capacity (DRC) plays an important role in genetic susceptibility to cancer. Polymorphisms of a number of DNA repair genes involved in several distinct pathways have been identified. However, their effects on repair function have not been well characterized. We demonstrated previously that DRC for removal of benzo[a]pyrene diol epoxide-induced DNA damage measured by a host-cell reactivation assay was modulated by two XPD/ERCC2 polymorphisms in lung cancer. In this report, we investigated the association between the repair phenotype of ultraviolet (UV)-induced damage and genotypes of three DNA repair genes, XPC and XPD [involved in nucleotide excision repair (NER)] and XRCC1 [involved in base excision repair (BER)]. We measured DRC for removal of UV photoproducts by the host-cell reactivation assay in cryopreserved lymphocytes from 102 healthy non Hispanic white subjects. We also typed these subjects for five polymorphisms in these three DNA repair genes (at intron 9 of XPC; exons 6, 10 and 23 of XPD and exon 10 of XRCC1). Compared with wild-type homozygotes, subjects homozygous for polymorphisms of the two NER genes consistently had suboptimal DRC. The DRC was consistently lower in subjects homozygous for XPC, XPD or both than in subjects with other genotypes, although the difference was not statistically significant for XPD variants. In contrast, the polymorphic allele of the BER gene, XRCC1, had no consistent effect on DRC. We concluded that these NER polymorphisms may modulate DRC and may be useful biomarkers for identifying individuals at risk of developing cancer. PMID- 11872634 TI - Expression of cyclin D1/2 in the lungs of strain A/J mice fed chemopreventive agents. AB - Male strain A mice were fed a diet containing chemopreventive agents. After 1 and 3 weeks on the diets, lung nuclear fractions were examined for expression of cyclin D1/2 with western blot analysis. In animals fed a diet containing a mixture of myoinositol and dexamethasone, a treatment found previously to be effective in preventing the development of tobacco smoke-induced lung tumors in A/J mice, cyclin D1/2 expression was reduced to 30-40% of control levels. A similar decrease in cyclin D1/2 expression was found when animals were fed either myoinositol or dexamethasone alone. Paradoxically, tobacco smoke by itself had a similar effect on cyclin D1/2 expression. On the other hand, several agents that had been previously found not to be effective against tobacco smoke carcinogenesis [phenethyl isothiocyanate, 1,4 phenylenebis(methylene)selenoisocyanate, N-acetylcysteine, acetylsalicylic acid, D-limonene and beta carotene] did not decrease cyclin D1/2 expression after 1 or 3 weeks of feeding. It was concluded that expression of cyclin D1/2 might be a potentially useful marker in the identification of chemopreventive agents for tobacco smoke and could be of some help in the evaluation of their effects. PMID- 11872637 TI - The in vivo levels of DNA alkylation products in human lymphocytes are not age dependent: an assay of 7-methyl- and 7-(2-hydroxyethyl)-guanine DNA adducts. AB - Endogenous DNA damage is assumed to be a major contributor to aging and cancer. This study compares the steady-state levels of 7-methyl- and 7-(2-hydroxyethyl) guanine DNA adducts in lymphocytes isolated from the younger (mean age 39.8 years) and the older (mean age 82.8 years) healthy subjects. Using a 32P-post labelling method, these adducts were measured in lymphocyte DNA from a total of 34 subjects. The results show that the amount of both 7-methyl- and 7-(2 hydroxyethyl)-guanine adducts in the younger age group was similar to that in the older age group. Our findings show that at steady-state the levels of DNA alkylation products are independent of age, suggesting that endogenous DNA damage, through methylation or lipid peroxidation, and the repair of such damage may not be deficient in lymphocytes of older individuals. PMID- 11872636 TI - Aromatic DNA adducts and polymorphisms of CYP1A1, NAT2, and GSTM1 in breast cancer. AB - Previous studies by us and others have shown a significantly higher level of aromatic DNA adducts in normal adjacent breast tissue samples obtained from breast cancer patients than in those obtained from non-cancerous controls. The increased amount of DNA damage could be related to excess environmental carcinogen exposure and/or genetic susceptibility to such exposure. In the current study, we investigated the relationship between the levels of aromatic DNA adducts in breast tissues and polymorphisms of the drug-metabolizing genes cytochrome P4501A1 (CYP1A1), N-acetyltransferase-2 (NAT2), and glutathione S transferase M1 (GSTM1), in 166 women having breast cancer. DNA adducts were measured using (32)P-postlabeling and information on smoking status was obtained from medical records. When pooled data of smokers and non-smokers were analyzed by multiple regression analyses, no significant correlation was found between the level of total DNA adducts and age, race, or polymorphisms of CYP1A1, GSTM1, and NAT2. The only significant predictor of the level of DNA adducts in breast tissues was smoking (P = 0.008). When data were analyzed separately in smokers and non-smokers, however, a significant gene-environment interaction was observed. Smokers with CYP1A1*1/*2 or *2/*2 genotypes had a significantly higher level of DNA adducts than those with the CYP1A1*1/*1 genotype. This effect was not seen among non-smokers. There was also a gene-gene interaction, as smokers with combined CYP1A1*1/*2 or CYP1A1*2/*2 genotypes and GSTM1 null had a much higher level of adducts than those with either CYP1A1 or GSTM1 polymorphism. Genetic polymorphisms of CYP1A1 and NAT2 were also significantly correlated with the frequency of certain types of DNA adducts. For example, a bulky benzo[a]pyrene (B[a]P)-like adduct was detected in 26% of the samples, the presence of which was not related to age, race, smoking status, or GSTM1 and NAT2 genotype. However, a significantly higher frequency of the B[a[P-like adduct was found in individuals having CYP1A1*1/*2 or *2/*2 genotypes than in those having the *1/*1 genotype (P = 0.04). In addition, individuals having slow NAT2 alleles had a significantly higher frequency of the typical smoking-related DNA adduct pattern, i.e. a diagonal radioactive zone (DRZ), than others did (P = 0.008). These findings suggest that polymorphisms of CYP1A1, GSTM1, and NAT2 significantly affect either the frequency or the level of DNA adducts in normal breast tissues of women having breast cancer, especially in smokers. Further large-scale studies are required to determine the exact role of these polymorphisms and types of DNA damage in breast cancer susceptibility. PMID- 11872638 TI - Associations between common polymorphisms in TP53 and p21WAF1/Cip1 and phenotypic features of breast cancer. AB - The tumour suppressor gene TP53 and its downstream effector p21 are thought to play major roles in the development of breast cancer. We investigated three common sequence variants in TP53 and p21 for possible associations with the risk of breast cancer and with various phenotypic features of this disease. A total of 351 cases were available for study. Germline DNA obtained from female subjects of similar age but without cancer was used to estimate the TP53 and p21 genotype frequencies in a control population. A single nucleotide polymorphism in intron 2 of p21 was associated with slightly increased breast cancer risk (RR = 1.4, P = 0.011) and with well/moderately differentiated tumour histology (P = 0.029). The 16 bp insertion polymorphism in intron 3 of TP53 was associated with poor histological grade (OR = 2.3, P = 0.013) independently of other pathological features. The codon 31 polymorphism in p21 was strongly linked to negative progesterone receptor status (OR = 3.4, P = 0.0001), suggesting this variant may have functional significance for the progesterone signalling pathway in breast cancer. These results add to the growing body of evidence that genetic variants can influence not only the risk of breast cancer but also the disease phenotype. PMID- 11872640 TI - Enhanced growth of colorectal aberrant crypt foci in fasted/refed rats involves changes in TGFbeta1 and p21CIP expressions. AB - We previously demonstrated that fasting/refeeding enhances the initiation phase of liver and colorectal carcinogenesis in rats. The present study was undertaken to establish whether cycles of fasting/refeeding carried out during the promotion phase of carcinogenesis may also affect the formation of aberrant crypt foci (ACF), preneoplastic lesions induced in the colon by azoxymethane (AOM). We were also interested in studying whether this effect might be mediated by changes in the proliferation, apoptosis or expression of TGFbeta1 and p21CIP genes in the colon. 44 male Fisher 344 rats were given a single dose of AOM (20 mg/kg s.c.) and one week later, they were exposed to 5 cycles of 4 days fasting followed by 7 10 days of refeeding (refed rats); controls were regularly fed; the rats were killed 2, 8 or 30 days after the last cycle of fasting. Fasting/refeeding caused a dramatic increase in crypt multiplicity when compared with regularly fed rats (AC/ACF was 4.30 +/- 1.3 in refed and 2.38 +/- 0.4 in regularly fed rats, P < 0.005 means +/- SD), while no significant changes were observed in the number of ACF/colon. In the two experimental groups, cell proliferation was higher in ACF than in the surrounding mucosa, but proliferative indexes were higher and the apoptotic index lower in ACF of refed rats compared with regularly fed rats. TGFbeta1 expression was higher in the ACF of refed rats than in those of fully fed controls while p21CIP was less expressed in refed rats than in controls. These results suggest that fasting/refeeding is a risk factor for colon cancer and must be taken into account for cancer prevention in humans. PMID- 11872639 TI - Effects of the isoflavone 4',5,7-trihydroxyisoflavone (genistein) on psoralen plus ultraviolet A radiation (PUVA)-induced photodamage. AB - Long-term psoralen plus ultraviolet A radiation (PUVA) therapy is associated with an increased risk of squamous cell carcinoma and malignant melanoma. Genistein (4',5,7-trihydroxyisoflavone), a major isoflavone in soybeans and a specific inhibitor of protein tyrosine kinase, has been shown to inhibit UVB induced skin carcinogenesis in hairless mice. For this study we examined the protective effects of topical genistein on PUVA-induced photodamage. In two separate experiments, genistein in a dimethyl sulfoxide/acetone (1:9) solution was applied to SKH-1 female mice 1 h post 8-methoxy-psoralen dosing and 1 h prior to UVA irradiation. Application of genistein significantly decreased PUVA-induced skin thickening, and greatly diminished cutaneous erythema and ulceration in a dose dependent manner. Histological examination showed that PUVA treatment of mouse skin induced dramatic inflammatory changes throughout the epidermis; topical genistein prevented these changes without noticeable adverse effects. Cells containing cleaved poly(ADP-ribose) polymerase (PARP) and active caspase-3 were significantly increased in PUVA-treated skin (P < 0.05 and P < 0.0001, respectively) as compared with unexposed control skin. Topical genistein completely inhibited cleavage of PARP and caspase-3. Proliferating cell nuclear antigen (PCNA) positive cells were observed in suprabasal areas of the epidermis and were significantly decreased in PUVA-treated skin compared with both control samples and samples treated with PUVA plus topical genistein (P < 0.005). These results indicate that genistein protects the skin from PUVA-induced photodamage. PMID- 11872641 TI - Catechol estrogen metabolites and conjugates in different regions of the prostate of Noble rats treated with 4-hydroxyestradiol: implications for estrogen-induced initiation of prostate cancer. AB - Prostate carcinomas arise in 100% of Noble rats treated with estradiol and testosterone. We hypothesize that estrogens initiate prostate cancer mainly by formation of 4-catechol estrogens (CE), followed by their oxidation to catechol estrogen-3,4-quinones (CE-3,4-Q), which can react with DNA. To avoid cancer initiation, CE can be detoxified by catechol-O-methyltransferase (COMT), and CE 3,4-Q by conjugation with glutathione (GSH) or by reduction to CE, catalyzed by quinone reductase and/or cytochrome P450 reductase. To investigate the prostatic metabolism of estrogens, Noble rats were treated with the CE 4-hydroxyestradiol (4-OHE2) or estradiol-3,4-quinone (E2-3,4-Q), and CE metabolites and conjugates were analyzed in the four regions of the prostate, which differ in susceptibility to carcinoma formation. Following treatment of rats with 4-OHE2 (6 micromol/100 g body weight in 200 microl of trioctanoin/dimethylsulfoxide (4:1) by intraperitoneal injection) for 90 min, the non-susceptible ventral (VP) and anterior (AP) prostate had higher levels of 4-methoxyCE and GSH conjugates than the susceptible dorsolateral prostate (DLP) and periurethral prostate (PUP). After treatment with the same molar amount of E2-3,4-Q, the VP and AP contained more GSH conjugates, 4-CE and 4-methoxyCE than the susceptible DLP and PUP. These results suggest that prostate areas susceptible to carcinoma induction have less protection by COMT, GSH, and quinone reductase and/or cytochrome P450 reductase, favoring reaction of CE-3,4-Q with DNA, presumably to initiate cancer. PMID- 11872642 TI - Aberrant CpG island methylation of the p16(INK4a) and estrogen receptor genes in rat lung tumors induced by particulate carcinogens. AB - Recent studies by our laboratory indicate that the p16(INK4a) gene is frequently methylated in lung tumors induced by genotoxic carcinogens and that the frequency for methylation of the estrogen receptor alpha (ER) gene varies as a function of carcinogenic exposure. The purpose of the current investigation was to define the role of these two genes in lung tumors induced by the particulate carcinogens carbon black (CB), diesel exhaust (DE) or beryllium metal. Methylation of p16 was observed in 59 and 46% of DE and CB tumors, respectively. In contrast, the ER gene was inactivated in only 15% of DE or CB tumors. Methylation of the p16 and ER genes was very common (80 and 50%, respectively) in beryllium-induced lung tumors; both genes were methylated in 40% of the tumors. Bisulfite sequencing revealed dense methylation throughout exon 1 of the ER gene. The inhibitory effect of methylation on gene transcription was confirmed through RT-PCR expression studies in which p16 gene expression was 30-60-fold lower in methylated than unmethylated tumors. Residual expression in methylated tumors was consistent with contamination by stromal and inflammatory cells. Results indicate that tumors induced by these particulate carcinogens arise, in part, through inactivation of the p16 and ER genes. Furthermore, the inactivation of the p16 gene by these carcinogenic exposures supports a possible role for oxidative stress and inflammation in the etiology of human lung cancer. PMID- 11872643 TI - Formation of 8-hydroxydeoxyguanosine and cell-cycle arrest in the rat liver via generation of oxidative stress by phenobarbital: association with expression profiles of p21(WAF1/Cip1), cyclin D1 and Ogg1. AB - To evaluate the risk of exposure to so-called non-genotoxic chemicals and elucidate mechanisms underlying their promoting activity on rat liver carcinogenesis the formation of 8-hydroxy-2'-deoxyguanosine (8-OHdG), cytochrome P-450 (P-450) and hydroxyl radicals induction, DNA repair and alteration to cellular proliferation and apoptosis in the rat liver were investigated during 2 weeks of phenobarbital (PB) administration at a dose of 0.05%. Significant increase of hydroxyl radical levels by day 4 of PB exposure accompanied the accumulation of 8-OHdG in the nucleus and P-450 isoenzymes CYP2B1/2 and CYP3A2 in the cytoplasm of hepatocytes. Conspicuous elevation of 8-OHdG and apoptosis in the liver tissue were associated with reduction of the proliferating cell nuclear antigen (PCNA) index after 8 days of PB application. Thereafter, 8-OHdG levels decreased with an increase in mRNA expression for the 8-OHdG repair enzyme, DNA glycosylase 1 (Ogg1). Analysis with LightCycler quantitative 2-step RT-PCR demonstrated induction of cyclin D1 (CD1) and p21(WAF1/Cip1) mRNA expression on days 4 and 6, respectively, preceding marked elevation of PCNA and apoptotic indices. These results suggest that similar to genotoxic, non-genotoxic chemicals might induce reversible alteration to nuclear 8-OHdG in the rat liver after several days of continuous application; however, by a different mechanism. Increased 8-OHdG formation is caused by developing oxidative stress or apoptotic degradation of DNA and coordinated with enhanced expression of CD1 mRNA and cell proliferation, subsequent increase of p21(WAF1/Cip1) mRNA expression, cell-cycle arrest and apoptosis, while activation of 8-OHdG repair mechanisms contributes to protection of tissue against reactive oxygen species-induced cell death. PMID- 11872644 TI - Influence of transfection with connexin 26 gene on malignant potential of human hepatoma cells. AB - We investigated the effect of transfection with connexin (Cx) 26 gene on the malignant potential of PLC/PRF/5 hepatoma cells, observing changes in their morphological features, alpha-fetoprotein (AFP) expression, cell proliferation and apoptosis in vitro, and their tumor growth in vivo. Fluorescence-activated cell sorting (FACS) analysis showed that 10.6% of PLC/PRF/5 hepatoma cells transfected with Cx26 cDNA expressed excessive Cx26, and the spread of lucifer yellow was wider in the colony of stable transfectants (PLC/Cx26) after its microinjection than in control. Nucleo-cytoplasmic (N/C) ratio was significantly lower in PLC/Cx26 (P < 0.0001). Cell proliferation assay showed significantly lower numbers in PLC/Cx26 on day 10 after seeding than in control (P = 0.0039), and AFP level /10(5) cells was significantly lower in medium of PLC/Cx26 (P = 0.0039). The number of proliferating cell nuclear antigen (PCNA)-positive cells was less in PLC/Cx26 in vitro than in control (P = 0.0039), and single-stranded DNA (ssDNA)-positive cells were more abundant in the colony of PLC/Cx26 (P = 0.029). Tumor volume in SCID mice was significantly smaller in the group of PLC/Cx26 than in the control (P < 0.01) throughout the observation period, and tumor weight of PLC/Cx26 was significantly lower (P = 0.0019) week 9 after inoculation. Transfection with Cx26 cDNA inhibited dedifferentiation, suppressed cell proliferation, and apoptosis was induced. Tumor growth of PLC/Cx26 was retarded. These findings suggest that transfection with Cx26 gene into human hepatoma cells reduces their malignant potential. PMID- 11872645 TI - Targeted expression of spermidine/spermine N1-acetyltransferase increases susceptibility to chemically induced skin carcinogenesis. AB - The bovine keratin 6 gene promoter was used to target expression of spermidine/spermine N1-acetyltransferase (SSAT) to epidermal keratinocytes in the hair follicle of transgenic mice. K6-SSAT transgenic mice appeared to be phenotypically normal and were indistinguishable from normal littermates until subjected to a two-stage tumorigenesis protocol. For such tumorigenesis studies, mice were bred for six generations onto a tumor promoter resistant C57BL/6 background strain. K6-SSAT transgenic mice showed a 10-fold increase in the number of epidermal tumors that developed in response to a single application of 400 nmol of the tumor initiator 7,12-dimethylbenz[a]anthracene followed by twice weekly applications of 17 nmol of the tumor promoter 12-O-tetradecanoylphorbol-13 acetate for 19 weeks. Tumor samples from transgenic animals showed marked elevations in SSAT enzyme activity and SSAT protein levels compared with tumors from non-transgenic littermates, and the accompanying changes in putrescine and N1-acetylspermidine pools indicated activation of SSAT-mediated polyamine catabolism in transgenic animals. An unusually high number of tumors were shown both grossly and histologically to have progressed to carcinomas in this model and these occurred with an early latency and only in mice carrying the K6-SSAT transgene. These results suggest that activation of polyamine catabolism leading to increases in putrescine and N1-acetylspermidine may play a key role in chemically induced mouse skin neoplasia. PMID- 11872647 TI - False cell lines. PMID- 11872646 TI - Analysis of DNA adducts in rats exposed to pentachlorophenol. AB - Pentachlorophenol (PCP) is a widely used biocide that has been reported to be hepatocarcinogenic in mice. Its effects in rats are equivocal, but the liver clearly is not a target organ for carcinogenesis. The carcinogenic effects of PCP in mice may relate to reactive oxygen species generated during metabolism. PCP is known to increase the hydroxyl radical-derived DNA lesion, 8-oxodeoxyguanosine (ohdG), in the liver of exposed mice. To investigate whether the generation of oxidative DNA damage and direct DNA adducts may explain the species difference in carcinogenicity, we have analyzed ohdG in hepatic DNA from PCP-exposed rats. Rats were exposed acutely to PCP for 1 or 5 days. Tissues also were obtained from a 27 week interim sacrifice of the 2 year National Toxicology Program carcinogenesis bioassay. We used HPLC with electrochemical array detection for ohdG analysis. Single or 5 day exposure to PCP (up to 120 or 60 mg/kg/day, respectively) did not increase ohdG. Dietary exposure to 1000 p.p.m. PCP (equivalent to 60 mg/kg/day) for 27 weeks induced a 2-fold increase in ohdG (1.8 versus 0.91x10(-6) in controls). In parallel, formation of direct DNA adducts was analyzed by 32P-post labeling following nuclease P1 adduct enrichment. We detected two major DNA adducts with relative adduct labeling of 0.78x10(7) adducts per total nucleotides. One of these adducts was found to co-migrate with the adduct induced by the metabolite, tetrachloro-1,4-benzoquinone. We observed differences in DNA adduct formation between acute and chronic studies, with acute studies not inducing any detectable amount of DNA adducts. These results indicated that chronic, but not acute exposure to PCP increased ohdG and direct adducts in hepatic DNA. As the same exposure conditions that enhanced ohdG did not produce liver cancer in rats, the generation of reactive oxygen species, oxidative DNA damage and direct DNA adducts is not sufficient for the induction of hepatocarcinogenesis by PCP in the rat. PMID- 11872650 TI - Molecular detection of circulating beta-cells after islet transplantation. AB - Islet transplantation is a promising treatment for type 1 diabetes. However, islet grafts are submitted to multiple injuries, including immunosuppressive drug toxicity, hyperglycemia, hypoxia, unspecific inflammatory reactions, as well as allo- and autoimmune destruction. Therapeutic approaches to these damage mechanisms require early detection of islet injury, which is currently not feasible because of the lack of efficient markers. Based on the hypothesis of islet dissociation and release of islet cells into the circulation during islet injury, we designed a highly sensitive and specific molecular assay, able to detect two beta-cells per milliliter of venous blood by RT-PCR of insulin mRNA. We report that circulating beta-cells can be demonstrated up to 10 weeks after intraportal islet transplantation, as assessed after six islet grafts in four type 1 diabetic patients. Furthermore, our results suggest that the time during which circulating islet cells can be detected may depend on the graft environment and the immunosuppressive regimen. This test may allow better estimation of islet cell loss and identification of factors involved in islet graft injury. PMID- 11872651 TI - Elevation of cytotoxic lymphocyte gene expression is predictive of islet allograft rejection in nonhuman primates. AB - Hyperglycemia and increased insulin requirements are indicators of ongoing islet allograft rejection, but there are no methods to predict or confirm rejection. Elevation of cytotoxic lymphocyte (CL) gene expression in peripheral blood (PB) has been correlated with renal allograft rejection in humans, but no published study has assessed the utility of monitoring these markers as predictors of rejection before the onset of clinical symptoms. We have established quantitative real-time PCR methods to determine the levels of mRNA transcripts for the CL genes granzyme B (GB), perforin, and fas ligand in blood samples from rhesus and cynomolgus monkeys. Four rhesus monkeys with long-term islet allograft function were studied. Antirejection (anti-CD154) therapy was discontinued, and weekly PB samples were obtained to determine whether the levels of mRNA transcripts for CL genes correlated with and/or were predictive of islet allograft rejection, defined as a loss of C-peptide production. For all monkeys, elevation of CL gene expression preceded rejection by 83--197 days, with GB as the best predictor. Elevated mRNA levels were sustained for 2--2.5 months in three of four animals and 1 month in the other, thus suggesting that the testing of these parameters may have practical applications in clinical islet cell transplantation. PMID- 11872652 TI - Effect of AICAR treatment on glycogen metabolism in skeletal muscle. AB - AMP-activated protein kinase (AMPK) is proposed to stimulate fat and carbohydrate catabolism to maintain cellular energy status. Recent studies demonstrate that pharmacologic activation of AMPK and mutations in the enzyme are associated with elevated muscle glycogen content in vivo. Our purpose was to determine the mechanism for increased muscle glycogen associated with AMPK activity in vivo. AMPK activity and glycogen metabolism were studied in red and white gastrocnemius muscles from rats treated with 5-aminoimidazole-4-carboxamide ribonucleoside (AICAR) in vivo, and also in muscles incubated with AICAR in vitro. In vivo AICAR treatment reduced blood glucose and increased blood lactate compared with basal values. AICAR increased muscle alpha2 AMPK activity, glycogen, and glucose-6 phosphate concentrations. Glycogen synthase activity was increased in the red gastrocnemius but was decreased in the white gastrocnemius. Glycogen phosphorylase activity increased in both muscles, with an inhibition initially observed in the red gastrocnemius. In vitro incubation with AICAR activated alpha2 AMPK but had no effect on either glycogen synthase or glycogen phosphorylase. These results suggest that AICAR treatment does not promote glycogen accumulation in skeletal muscle in vivo by altering glycogen synthase and glycogen phosphorylase. Rather, the increased glycogen is due to the well known effects of AICAR to increase glucose uptake. PMID- 11872653 TI - Mode of transcapillary transport of insulin and insulin analog NN304 in dog hindlimb: evidence for passive diffusion. AB - A defect in transcapillary transport of insulin in skeletal muscle and adipose tissue has been proposed to play a role in the insulin resistance that leads to type 2 diabetes, yet the mechanism of insulin transfer across the capillary endothelium from plasma to interstitium continues to be debated. This study examined in vivo the interstitial appearance of insulin in hindlimb using the fatty acid acylated insulin analog Lys(B29)-tetradecanoyl des-(B30) human insulin, or NN304, as a marker for insulin transport. If the insulin transport were a saturable process, then "swamping" the capillary endothelial insulin receptors with native insulin would suppress the subsequent appearance in interstitial fluid of the insulin analog NN304. This analog binds to insulin receptors with an affinity of about 50% of native insulin. Experimental conditions established a physiologic NN304 dose in the absence or presence of pharmacologic and saturating concentrations of regular human insulin. Euglycemic clamps were performed in dogs under inhalant anesthesia with deep hindlimb lymphatic sampling, representative of skeletal muscle interstitial fluid (ISF). In group 1 (n = 8), NN304 alone was infused (3.6 pmol center dot min(-1) center dot kg(-1)) from 60 to 360 min. In group 2 (n = 6), starting at time 0, human insulin was infused at a pharmacologic dose (60 pmol center dot min(-1) center dot kg(-1)) with the addition of NN304 infusion (3.6 pmol center dot min(-1) center dot kg(-1)) from 60 to 360 min. In group 3 (n = 4), the human insulin infusion was increased to a saturating dose (120 pmol center dot min(-1) center dot kg(-1)). Pharmacologic insulin infusion (group 2) established steady-state human insulin concentrations of 6,300 plus minus 510 pmol/l in plasma and 5,300 plus minus 540 pmol/l in ISF. Saturating insulin infusion (group 3) achieved steady-state human insulin concentrations of 22,000 plus minus 1,800 pmol/l in plasma and 19,000 plus minus 1,500 pmol/l in ISF. Total (bound and unbound) NN304 plasma concentrations rose from a steady state of 1,900 plus minus 110 (group 1) to 2,400 plus minus 200 pmol/l (group 2) and 3,100 plus minus 580 pmol/l (group 3), consistent with a competition-driven decline in NN304 clearance from plasma as the human insulin level increased (P < 0.05 by ANOVA). Steady-state interstitial NN304 concentrations also rose with increasing human insulin levels but did not achieve significance in comparison with analog alone (162 plus minus 15 vs. 196 plus minus 22 and 241 plus minus 53 pmol/l for group 1 versus groups 2 and 3, respectively; P = 0.20), yet the steady-state plasma:ISF ratio for NN304 remained essentially unchanged in the absence and presence of elevated human insulin levels (12.6 plus minus 1.2 vs. 12.4 plus minus 0.5 and 13.1 plus minus 1.5 for group 1 versus groups 2 and 3, respectively; P = 0.93). Last, NN304 rate of appearance in interstitial fluid (i.e., half-time to steady state) was similar between groups; mean half-time of 92 plus minus 4 min (NS between groups). In conclusion, appearance of the insulin analog NN304 in skeletal muscle interstitial fluid was constant whether in the absence or presence of human insulin concentrations sufficient to saturate the endothelial insulin receptors. These findings support the hypothesis, provided that the mechanism of insulin and NN304 transcapillary transport is similar, that transcapillary transport of insulin in skeletal muscle occurs primarily via a nonsaturable process such as passive diffusion via a paracellular or transcellular route. PMID- 11872655 TI - Human skeletal muscle expresses a glycogen-targeting subunit of PP1 that is identical to the insulin-sensitive glycogen-targeting subunit G(L) of liver. AB - Insulin has been previously shown to regulate the expression of the hepatic glycogen-targeting subunit, G(L), of protein phosphatase 1 (PP1) and is believed to control the activity of the PP1-G(L) complex by modulation of the level of phosphorylase a, which allosterically inhibits the activity of PP1-G(L). These mechanisms contribute to the ability of insulin to increase hepatic glycogen synthesis. Human G(L) shows >88% amino acid identity to its rat and mouse homologs, with complete conservation of the phosphorylase a binding site. G(L) is highly expressed in the liver and present at appreciable levels in heart tissue of all three species. Surprisingly, G(L) is highly expressed in human skeletal muscle while only being detected at very low levels in rat, mouse, and rabbit skeletal muscle. The amino acid sequence of G(L) predicted from the cDNA is identical in human liver and skeletal muscle and encoded by a gene on chromosome 8 at p23.1. The species-specific difference in the level of expression of G(L) mRNA and protein in skeletal muscle has important implications for understanding the mechanisms by which insulin regulates glycogen synthesis in human skeletal muscle and for questions regarding whether rodents are appropriate models for this purpose. PMID- 11872654 TI - Caffeine-induced impairment of insulin action but not insulin signaling in human skeletal muscle is reduced by exercise. AB - We investigated the effects of caffeine ingestion on skeletal muscle glucose uptake, glycogen synthase (GS) activity, and insulin signaling intermediates during a 100-min euglycemic-hyperinsulinemic (100 microU/ml) clamp. On two occasions, seven men performed 1-h one-legged knee extensor exercise at 3 h before the clamp. Caffeine (5 mg/kg) or placebo was administered in a randomized, double-blind fashion 1 h before the clamp. During the clamp, whole-body glucose disposal was reduced (P < 0.05) in caffeine (37.5 +/- 3.1 micromol x min(-1) x kg(-1)) vs. placebo (54.1 +/- 2.9 micromol x min(-1) x kg(-1)). In accordance, the total area under the curve over 100 min (AUC(0--100 min)) for insulin stimulated glucose uptake in caffeine was reduced (P < 0.05) by approximately 50% in rested and exercised muscle. Caffeine also reduced (P < 0.05) GS activity before and during insulin infusion in both legs. Exercise increased insulin sensitivity of leg glucose uptake in both caffeine and placebo. Insulin increased insulin receptor tyrosine kinase (IRTK), insulin receptor substrate 1-associated phosphatidylinositol (PI) 3-kinase activities, and Ser(473) phosphorylation of protein kinase B (PKB)/Akt significantly but similarly in rested and exercised legs. Furthermore, insulin significantly decreased glycogen synthase kinase 3alpha (GSK-3alpha) activity equally in both legs. Caffeine did not alter insulin signaling in either leg. Plasma epinephrine and muscle cAMP concentrations were increased in caffeine. We conclude that 1) caffeine impairs insulin-stimulated glucose uptake and GS activity in rested and exercised human skeletal muscle; 2) caffeine-induced impairment of insulin-stimulated muscle glucose uptake and downregulation of GS activity are not accompanied by alterations in IRTK, PI 3 kinase, PKB/Akt, or GSK-3alpha but may be associated with increases in epinephrine and intramuscular cAMP concentrations; and 3) exercise reduces the detrimental effects of caffeine on insulin action in muscle. PMID- 11872656 TI - Mechanism of amino acid-induced skeletal muscle insulin resistance in humans. AB - Plasma concentrations of amino acids are frequently elevated in insulin-resistant states, and a protein-enriched diet can impair glucose metabolism. This study examined effects of short-term plasma amino acid (AA) elevation on whole-body glucose disposal and cellular insulin action in skeletal muscle. Seven healthy men were studied for 5.5 h during euglycemic (5.5 mmol/l), hyperinsulinemic (430 pmol/l), fasting glucagon (65 ng/l), and growth hormone (0.4 microg/l) somatostatin clamp tests in the presence of low (approximately 1.6 mmol/l) and increased (approximately 4.6 mmol/l) plasma AA concentrations. Glucose turnover was measured with D-[6,6-(2)H(2)]glucose. Intramuscular concentrations of glycogen and glucose-6-phosphate (G6P) were monitored using (13)C and (31)P nuclear magnetic resonance spectroscopy, respectively. A approximately 2.1-fold elevation of plasma AAs reduced whole-body glucose disposal by 25% (P < 0.01). Rates of muscle glycogen synthesis decreased by 64% (180--315 min, 24 plus minus 3; control, 67 plus minus 10 micromol center dot l(-1) center dot min(-1); P < 0.01), which was accompanied by a reduction in G6P starting at 130 min (DeltaG6P(260--300 min), 18 plus minus 19; control, 103 plus minus 33 micromol/l; P < 0.05). In conclusion, plasma amino acid elevation induces skeletal muscle insulin resistance in humans by inhibition of glucose transport/phosphorylation, resulting in marked reduction of glycogen synthesis. PMID- 11872657 TI - Fructose improves the ability of hyperglycemia per se to regulate glucose production in type 2 diabetes. AB - The ability of hyperglycemia per se to suppress endogenous glucose production (GP) is blunted in type 2 diabetes. This could be due in part to decreased glucose-induced flux through glucokinase (GK). Because fructose activates hepatic GK, we examined whether catalytic amounts of fructose could restore inhibition of GP by hyperglycemia in humans with type 2 diabetes. Glucose fluxes ([3 (3)H]glucose) were measured during euglycemia (5 mmol/l) and after abrupt onset of hyperglycemia (10 mmol/l; variable dextrose infusion) under fixed hormonal conditions (somatostatin infusion for 6 h with basal insulin/glucagon/growth hormone replacement). A total of 10 subjects with moderately controlled type 2 diabetes and 7 age- and BMI-matched nondiabetic subjects were studied on up to three separate occasions under the following conditions: without fructose (F(-)) or with infusion of fructose at two dosages: 0.6 mg/kg center dot min (low F) and 1.8 mg/kg center dot min (high F). Although GP failed to decrease in response to hyperglycemia in type 2 diabetes, the coinfusion of both doses of fructose was associated with comparable decreases in GP in response to hyperglycemia (low F = 27%, high F = -33%; P < 0.01 vs. F(-) at both dosages), which approached the 44% decline in GP observed without fructose in the nondiabetic subjects. GP responses to hyperglycemia were not altered by the addition of fructose in the nondiabetic group (low F = -47%, high F = -42%; P > 0.05 vs. F(-)). Thus, the administration of small amounts of fructose to type 2 diabetic subjects partially corrected the regulation of GP by hyperglycemia per se, yet did not affect this regulation in the nondiabetic subjects. This suggests that the liver's inability to respond to hyperglycemia in type 2 diabetes, likely caused by impaired GK activity, contributes substantially to the increased GP in these individuals. PMID- 11872659 TI - Increased fatty acid re-esterification by PEPCK overexpression in adipose tissue leads to obesity without insulin resistance. AB - Adipose tissue glyceroneogenesis generates glycerol 3-phosphate, which could be used for fatty acid esterification during starvation. To determine whether increased glyceroneogenesis leads to increased fat mass and to explore the role of obesity in the development of insulin resistance, we overexpressed PEPCK, a regulatory enzyme of glyceroneogenesis in adipose tissue. Transgenic mice showed a chronic increase in PEPCK activity, which led to increased glyceroneogenesis, re-esterification of free fatty acids (FFAs), increased adipocyte size and fat mass, and higher body weight. In spite of increased fat mass, transgenic mice showed decreased circulating FFAs and normal leptin levels. Moreover, glucose tolerance and whole-body insulin sensitivity were preserved. Skeletal muscle basal and insulin-stimulated glucose uptake and glycogen content were not affected, suggesting that skeletal muscle insulin sensitivity is normal in transgenic obese mice. Our results indicate the key role of PEPCK in the control of FFA re-esterification in adipose tissue and, thus, the contribution of glyceroneogenesis to fat accumulation. Moreover, they suggest that higher fat mass without increased circulating FFAs does not lead to insulin resistance or type 2 diabetes in these mice. PMID- 11872658 TI - Insulin and isoproterenol differentially regulate mitogen-activated protein kinase-dependent Na(+)-K(+)-2Cl(-) cotransporter activity in skeletal muscle. AB - Recent studies have demonstrated that p44/42(MAPK) extracellular signal-regulated kinase (ERK)1 and -2-dependent Na(+)-K(+)-2Cl(-) co-transporter (NKCC) activity may contribute to total potassium uptake by skeletal muscle. To study the precise mechanisms regulating NKCC activity, rat soleus and plantaris muscles were stimulated ex vivo by insulin or isoproterenol (ISO). Both hormones stimulated total uptake of the potassium congener (86)Rb by 25--70%. However, only ISO stimulated the NKCC-mediated (86)Rb uptake. Insulin inhibited the ISO-stimulated NKCC activity, and this counteraction was sensitive to the p38 mitogen-activated protein kinase (MAPK) inhibitor SB203580 in the predominantly slow-twitch soleus muscle. Pretreatment of the soleus muscle with the phosphatidylinositol (PI) 3 kinase inhibitors wortmannin and LY294002 or with SB203580 uncovered an insulin stimulated NKCC activity and also increased the insulin-stimulated phosphorylation of ERK. In the predominantly fast-twitch plantaris muscle, insulin-stimulated NKCC activity became apparent only after inhibition of PI 3 kinase activity, accompanied by an increase in ERK phosphorylation. PI 3-kinase inhibitors also abolished insulin-stimulated p38 MAPK phosphorylation in the plantaris muscle and Akt phosphorylation in both muscles. These data demonstrated that insulin inhibits NKCC-mediated transport in skeletal muscle through PI 3 kinase-sensitive and SB203580-sensitive mechanisms. Furthermore, differential activation of signaling cascade elements after hormonal stimulation may contribute to fiber-type specificity in the control of potassium transport by skeletal muscle. PMID- 11872660 TI - C(2)-ceramide influences the expression and insulin-mediated regulation of cyclic nucleotide phosphodiesterase 3B and lipolysis in 3T3-L1 adipocytes. AB - Cyclic nucleotide phosphodiesterase (PDE) 3B plays an important role in the antilipolytic action of insulin and, thereby, the release of fatty acids from adipocytes. Increased concentrations of circulating fatty acids as a result of elevated or unrestrained lipolysis cause insulin resistance. The lipolytic action of tumor necrosis factor (TNF)-alpha is thought to be one of the mechanisms by which TNF-alpha induces insulin resistance. Ceramide is the suggested second messenger of TNF-alpha action, and in this study, we used 3T3-L1 adipocytes to investigate the effects of C(2)-ceramide (a short-chain ceramide analog) on the expression and regulation of PDE3B and lipolysis. Incubation of adipocytes with 100 micromol/l C(2)-ceramide (N-acetyl-sphingosine) resulted in a time-dependent decrease of PDE3B activity, accompanied by decreased PDE3B protein expression. C(2)-ceramide, in a time- and dose-dependent manner, stimulated lipolysis, an effect that was blocked by H-89, an inhibitor of protein kinase A. These ceramide effects were prevented by 20 micromol/l troglitazone, an antidiabetic drug. In addition to downregulation of PDE3B, the antilipolytic action of insulin was decreased by ceramide treatment. These results, together with data from other studies on PDE3B and lipolysis in diabetic humans and animals, suggest a novel pathway by which ceramide induces insulin resistance. Furthermore, PDE3B is demonstrated to be a target for troglitazone action in adipocytes. PMID- 11872662 TI - Genetic risk determines the emergence of diabetes-associated autoantibodies in young children. AB - Timing of onset of autoimmunity is a prerequisite for unmasking triggers and pathogenesis of type 1 diabetes. We followed 4,590 consecutive newborns with 8 or 3% HLA-DQB1 conferred risk for type 1 diabetes at 3-, 6-, or 12-month intervals up to 5.5 years of age. Islet cell autoantibodies (ICAs) and, in the 137 children with ICAs, insulin autoantibodies (IAAs), GAD65 autoantibodies (GADAs), and IA-2 protein autoantibodies (IA-2As) were measured. Children with high genetic risk developed ICAs more often than those with moderate risk (log-rank P = 0.0015); 85 and 91% remained ICA negative by 5 years of age, respectively. The time of appearance of biochemical autoantibodies was then compared with the appearance of ICAs. IAAs and GADAs emerged usually before ICAs (means -1.8 and -1.5 months, respectively) and IA-2As after ICAs (mean 2.0 months). Ninety-five percent of all IAAs, GADAs, and IA-2As seroconversions occurred in a cluster (-12 to 8 months) around the ICA seroconversion. We conclude that diabetes-associated autoantibodies emerged in children with predisposing HLA-DQB1 alleles after 3 months of age at a constant tempo, determined by the genetic risk level, usually in the order of IAA, GADA, ICA, and IA-2A. Seroconversion to multiple autoantibody positivity usually occurred tightly clustered in time. PMID- 11872661 TI - Combination therapy with sirolimus and interleukin-2 prevents spontaneous and recurrent autoimmune diabetes in NOD mice. AB - Sirolimus is an immunosuppressant that inhibits interleukin (IL)-2 signaling of T cell proliferation but not IL-2-induced T-cell apoptosis. Therefore, we hypothesized that administration of IL-2, together with sirolimus, might shift T cell proliferation to apoptosis and prevent autoimmune destruction of islet beta cells. We found that sirolimus and IL-2 therapy of female NOD mice, beginning at age 10 weeks, was synergistic in preventing diabetes development, and disease prevention continued for 13 weeks after stopping sirolimus and IL-2 therapy. Similarly, sirolimus and IL-2 were synergistic in protecting syngeneic islet grafts from recurrent autoimmune destruction after transplantation in diabetic NOD mice, and diabetes did not recur after stopping sirolimus and IL-2 combination therapy. Immunocytochemical examination of islet grafts revealed significantly decreased numbers of leukocytes together with increased apoptosis of these cells in mice treated with sirolimus and IL-2, whereas beta-cells were more numerous, and significantly fewer were apoptotic. In addition, Th1-type cells (gamma-interferon-positive and IL-2(+)) were decreased the most, and Th2 type cells (IL-4(+) and IL-10(+)) and Th3-type cells (transforming growth factor beta1(+)) were increased the most in islet grafts of sirolimus and IL-2-treated mice. We conclude that 1) combination therapy with sirolimus and IL-2 is synergistic in protecting islet beta-cells from autoimmune destruction; 2) diabetes prevention continues after withdrawal of therapy; and 3) the mechanism of protection involves a shift from Th1- to Th2- and Th3-type cytokine-producing cells, possibly due to deletion of autoreactive Th1 cells. PMID- 11872663 TI - Dipeptidyl peptidase IV-resistant [D-Ala(2)]glucose-dependent insulinotropic polypeptide (GIP) improves glucose tolerance in normal and obese diabetic rats. AB - The therapeutic potential of glucose-dependent insulinotropic polypeptide (GIP) for improving glycemic control has largely gone unstudied. A series of synthetic GIP peptides modified at the NH(2)-terminus were screened in vitro for resistance to dipeptidyl peptidase IV (DP IV) degradation and potency to stimulate cyclic AMP and affinity for the transfected rat GIP receptor. In vitro experiments indicated that [D-Ala(2)]GIP possessed the greatest resistance to enzymatic degradation, combined with minimal effects on efficacy at the receptor. Thus, [D Ala(2)]GIP(1--42) was selected for further testing in the perfused rat pancreas and bioassay in conscious Wistar and Zucker rats. When injected subcutaneously in normal Wistar, Fa/?, or fa/fa Vancouver Diabetic Fatty (VDF) Zucker rats, both GIP and [D-Ala(2)]GIP significantly reduced glycemic excursions during a concurrent oral glucose tolerance test via stimulation of insulin release. The latter peptide displayed greater in vivo effectiveness, likely because of resistance to enzymatic degradation. Hence, despite reduced bioactivity in diabetic models at physiological concentrations, GIP and analogs with improved plasma stability still improve glucose tolerance when given in supraphysiological doses, and thus may prove useful in the treatment of diabetic states. PMID- 11872664 TI - Differential effects of hyperlipidemia on insulin secretion in islets of langerhans from hyperglycemic versus normoglycemic rats. AB - Chronic elevations in plasma levels of fatty acids (FAs) adversely affect pancreatic beta-cell function in type 2 diabetes. In vitro, we have previously shown that deleterious effects of prolonged exposure of isolated islets to FAs were dependent on the presence of elevated glucose concentration. This led us to hypothesize that both hyperlipidemia and hyperglycemia must be present simultaneously for FAs to affect beta-cell function. To test this hypothesis in vivo, we administered a high-fat diet for 6 weeks to Goto-Kakizaki (GK) rats. High-fat feeding had no effect on insulin secretion, insulin content, or insulin mRNA levels in islets from normoglycemic Wistar rats. In contrast, high-fat feeding markedly impaired glucose-induced insulin secretion in islets from GK rats. High-fat feeding did not affect triglyceride (TG) content or the rate of glucose oxidation in islets. It was, however, accompanied by a twofold increase in uncoupling protein (UCP)-2 levels in GK rat islets. Insulin treatment completely normalized glucose-induced insulin secretion and prevented the increase in UCP-2 expression in islets from high-fat-fed GK rats. We conclude that hyperlipidemia induced by high-fat feeding affects insulin secretion in islets from hyperglycemic GK rats only, by a mechanism which may involve, at least in part, modulation of UCP-2 expression. PMID- 11872665 TI - Assessment of the role of interstitial glucagon in the acute glucose secretory responsiveness of in situ pancreatic beta-cells. AB - Glucagon is a potent stimulator of insulin release in the presence of a permissive glucose concentration, activating beta-cells in vitro via both glucagon- and glucagon-like peptide-1 (GLP-1)-receptors. It is still unclear whether locally released glucagon amplifies the secretory responsiveness of neighboring beta-cells in the intact pancreas. The present study investigates this question in the perfused pancreas by examining the effects of antagonists for glucagon receptors ([des-His(1),des-Phe(6),Glu(9)]glucagon-NH(2), 10 micromol/l) and GLP-1-receptors [exendin-(9-39)-NH(2), 1 micromol/l] on the insulin secretory response to glucose. The specificity of both antagonists was demonstrated by their selective interaction with glucagon-receptor signaling in rat hepatocytes and GLP-1-receptor signaling in Chinese hamster lung (CHL) fibroblasts. In purified rat beta-cells, the glucagon-receptor antagonist (10 micromol/l) inhibited the effect of 1 nmol/l glucagon upon glucose-induced insulin release by 78 plus minus 6%. In the perfused rat pancreas, neither of these antagonists inhibited the potent secretory response to 20 mmol/l glucose, although they effectively suppressed the potentiating effect of, respectively, an infusion of glucagon (1 nmol/l) or GLP-1 (1 nmol/l) on insulin release. When endogenous glucagon release was enhanced by isoproterenol (100 nmol/l), no amplification was seen in the simultaneous or subsequent insulin secretory response to glucose. It is concluded that, at least under the present selected conditions, the glucose-induced insulin release by the perfused rat pancreas seems to occur independent of an amplifying glucagon signal from neighboring alpha-cells. PMID- 11872666 TI - Peroxisomal proliferator-activated receptor-gamma upregulates glucokinase gene expression in beta-cells. AB - Thiazolidinediones, synthetic ligands of peroxisomal proliferator-activated receptor-gamma (PPAR-gamma), improve peripheral insulin sensitivity and glucose stimulated insulin secretion in pancreatic beta-cells. To explore the role of PPAR-gamma in glucose sensing of beta-cells, we have dissected the beta-cell specific glucokinase (betaGK) promoter, which constitutes glucose-sensing apparatus in pancreatic beta-cells, and identified a peroxisomal proliferator response element (PPRE) in the promoter. The betaGK-PPRE is located in the region between +47 and +68 bp. PPAR-gamma/retinoid X receptor-alpha heterodimer binds to the element and activates the betaGK promoter. The betaGK promoter lacking or having mutations in PPRE cannot be activated by PPAR-gamma. PPAR-gamma activates the betaGK promoter in beta-cells as well as non-beta-cells. Furthermore, troglitazone increases endogenous GK expression and its enzyme activity in beta cell lines. These results indicate that PPAR-gamma can regulate GK expression in beta-cells. Taking these results together with our previous work, we conclude that PPAR-gamma regulates gene expression of glucose-sensing apparatus and thereby improves glucose-sensing ability of beta-cells, contributing to the restoration of beta-cell function in type 2 diabetic subjects by troglitazone. PMID- 11872667 TI - Gastrin stimulates beta-cell neogenesis and increases islet mass from transdifferentiated but not from normal exocrine pancreas tissue. AB - It is still unclear which factors regulate pancreatic regeneration and beta-cell neogenesis and which precursor cells are involved. We evaluated the role of intravenously infused gastrin in regenerating pancreas of duct-ligated rats. The ligation of exocrine ducts draining the splenic half of the pancreas resulted in acinoductal transdifferentiation within the ligated part but not in the unligated part. We found that infusion of gastrin from day 7 to 10 postligation resulted in a doubling of the beta-cell mass in the ligated part as measured by morphometry. This increase in insulin-expressing cells was not associated with increased proliferation, hypertrophy, or reduced cell death of the beta-cells. Furthermore, we found an increased percentage of single, extra-insular beta-cells and small beta-cell clusters induced by gastrin infusion. These changes occurred only in the ligated part of the pancreas, where transdifferentiation of the exocrine acinar cells to ductlike cells (metaplasia) had occurred, and was not found in the normal unaffected pancreatic tissue. In conclusion, we demonstrate that administration of gastrin stimulates beta-cell neogenesis and expansion of the beta-cell mass from transdifferentiated exocrine pancreas. PMID- 11872668 TI - NFAT regulates insulin gene promoter activity in response to synergistic pathways induced by glucose and glucagon-like peptide-1. AB - Currently there is intense interest to define the mechanism of action of glucagon like peptide-1 (GLP-1) in regulating beta-cell function, including insulin gene transcription. In this study, GLP-1 (100 nmol/l), in the presence of glucose (11 mmol/l), induced a similar71-fold increase in insulin gene promoter activity in INS-1 pancreatic beta-cells, an effect that was an order of magnitude larger than with either stimulant alone. The response to GLP-1 was mimicked by forskolin and largely inhibited by the protein kinase A (PKA) inhibitors, H89 and myristoylated PKI(14--22) amide, indicating partial mediation via a cAMP/PKA pathway. Significantly, the actions of both GLP-1 and forskolin were abolished by the selective Ca(2+)/calmodulin-dependent phosphatase 2B (calcineurin) inhibitor, FK506, as well as by the chelation of intracellular Ca(2+) by BAPTA (bis(2 aminophenoxy)ethane-N,N,N',N'-tetraacetate). Glucose and GLP-1 also synergistically activated NFAT (nuclear factor of activated T-cells)-mediated transcription from a minimal promoter construct containing tandem NFAT consensus sequences. Furthermore, two-point base pair mutations in any of the three identified NFAT sites within the rat insulin I promoter resulted in a significant reduction in the combined effect of glucose and GLP-1. These data suggest that the synergistic action of glucose and GLP-1 to promote insulin gene transcription is mediated through NFAT via PKA- and calcineurin-dependent pathways in pancreatic beta-cells. PMID- 11872669 TI - Primary in vivo oscillations of metabolism in the pancreas. AB - The role of metabolism in the generation of plasma insulin oscillations was investigated by simultaneous in vivo recordings of oxygen tension (pO(2)) in the endocrine and exocrine pancreas and portal blood insulin concentrations in the anesthetized rat. At the start of the experiment, the blood glucose concentration of seven rats was 6.2 +/- 0.1 mmol/l and the arterial blood pressure was 116 +/- 5 mmHg. These values did not differ from those obtained at the end of the experiment. Islet pO(2) was measured by impaling superficially located islets with a miniaturized Clark electrode. The pO(2) measurements revealed slow (0.21 +/- 0.03 min(-1)) with superimposed rapid (3.1 +/- 0.3 min(-1)) oscillations. The average pO(2) was 39 +/- 5 mmHg. Simultaneous recordings of pO(2) in the exocrine pancreas were significantly lower (16 +/- 6 mmHg), but showed a slow and a rapid oscillatory activity with similar frequencies as seen in the endocrine pancreas. Corresponding measurements of portal insulin concentrations revealed insulin oscillations at a frequency of 0.22 +/- 0.02 min(-1). The results are the first in vivo recordings of an oscillatory islet parameter with a frequency corresponding to that of plasma insulin oscillations; they support a primary role of metabolic oscillations in the induction of plasma insulin oscillations. PMID- 11872670 TI - Counteraction of type 1 diabetic alterations by engineering skeletal muscle to produce insulin: insights from transgenic mice. AB - Insulin replacement therapy in type 1 diabetes is imperfect because proper glycemic control is not always achieved. Most patients develop microvascular, macrovascular, and neurological complications, which increase with the degree of hyperglycemia. Engineered muscle cells continuously secreting basal levels of insulin might be used to improve the efficacy of insulin treatment. Here we examined the control of glucose homeostasis in healthy and diabetic transgenic mice constitutively expressing mature human insulin in skeletal muscle. Fed transgenic mice were normoglycemic and normoinsulinemic and, after an intraperitoneal glucose tolerance test, showed increased glucose disposal. When treated with streptozotocin (STZ), transgenic mice showed increased insulinemia and reduced hyperglycemia when fed and normoglycemia and normoinsulinemia when fasted. Injection of low doses of soluble insulin restored normoglycemia in fed STZ-treated transgenic mice, while STZ-treated controls remained highly hyperglycemic, indicating that diabetic transgenic mice were more sensitive to the hypoglycemic effects of insulin. Furthermore, STZ-treated transgenic mice presented normalization of both skeletal muscle and liver glucose metabolism. These results indicate that skeletal muscle may be a key target tissue for insulin production and suggest that muscle cells secreting basal levels of insulin, in conjunction with insulin therapy, may permit tight regulation of glycemia. PMID- 11872671 TI - Unregulated elevation of glutamate dehydrogenase activity induces glutamine stimulated insulin secretion: identification and characterization of a GLUD1 gene mutation and insulin secretion studies with MIN6 cells overexpressing the mutant glutamate dehydrogenase. AB - Glutamate dehydrogenase (GDH) is important in normal glucose homeostasis. Mutations of GDH result in hyperinsulinism/hyperammonemia syndrome. Using PCR/single-strand conformation polymorphism analysis of the gene encoding GDH in 12 Japanese patients with persistent hyperinsulinemic hypoglycemia of infancy (PHHI), we found a mutation (Y266C) in one PHHI patient. This mutation was not found in any of the control or type 2 diabetic subjects. The activity of the mutant GDH (GDH266C), expressed in COS-7 cells, was constitutively elevated, and allosteric regulations by ADP and GTP were severely impaired. The effect of the unregulated increase in GDH activity on insulin secretion was examined by overexpressing GDH266C in an insulinoma cell line, MIN6. Although glutamine alone did not stimulate insulin secretion from control MIN6-lacZ, it remarkably stimulated insulin secretion from MIN6-GDH266C. This finding suggests that constitutively activated GDH enhances oxidation of glutamate, which is intracellularly converted from glutamine to alpha-ketoglutarate, a tricarboxylic acid cycle substrate, which thereby stimulates insulin secretion. Interestingly, insulin secretion is also exaggerated significantly at low glucose concentrations (2 and 5 mmol/l) but not at higher glucose concentrations (8--25 mmol/l). Our results directly illustrate the importance of GDH in the regulation of insulin secretion from pancreatic beta-cells. PMID- 11872672 TI - Regional differences in the response of human pre-adipocytes to PPARgamma and RXRalpha agonists. AB - We have previously reported that omental (OM) preadipocytes respond less well to the prodifferentiating effects of thiazolidinediones than do preadipocytes from subcutaneous (SC) depots. This finding is consistent with in vivo alterations in fat distribution that occur in humans treated with thiazolidinediones. To explore these site-related differences further, we used real-time RT-PCR to quantify the specific mRNAs encoding peroxisome proliferator-activated receptor (PPAR) gamma1 and gamma2 and found that both isoforms were more highly expressed in SC than in OM preadipocytes. After 10 days of thiazolidinedione treatment, preadipocytes from both depots showed a small and comparable increase in expression of PPARgamma1 mRNA (1.7 +/- 0.2-fold [P = 0.007]) and 1.3 +/- 0.1-fold [P = 0.008] increase for SC and OM, respectively). There was a much larger increase in PPARgamma2 expression, which was significantly greater in SC compared with OM preadipocytes (11.1 +/- 2.8-fold [P = 0.0003] and 5.5 +/- 1.7-fold [P = 0.0003], respectively; P = 0.014 for SC versus OM). To establish whether the refractoriness of OM preadipocytes to differentiation was unique to activators of the PPARgamma pathway, we examined the effects of the retinoid X receptor (RXR) ligand LG100268. As assessed by glycerol-3-phosphate dehydrogenase activity, LG100268 had a greater effect on the differentiation of SC compared with OM preadipocytes when examined alone (SC = 5.7 +/- 1.7-fold vs. OM = 1.9 +/- 0.6 fold; P < 0.05) or in combination with rosiglitazone (SC = 27.0 +/- 7.5 vs. OM = 10.6 +/- 3.6-fold; P < 0.05). Consistent with this, RXRalpha mRNA levels were also higher in SC than in OM preadipocytes. In summary, the previously reported insensitivity of OM preadipocytes to the differentiating effects of thiazolidinediones may relate to their lower basal levels of PPARgamma1 and gamma2 mRNA and their diminished capacity to upregulate PPARgamma2 expression in response to ligand. That omentally derived cells also show reduced responsiveness to the prodifferentiating actions of an RXR ligand and a lower expression of RXRalpha in the undifferentiated state suggests that they may have a more generalized resistance to differentiation. PMID- 11872673 TI - Hypoglycemia-associated autonomic failure in advanced type 2 diabetes. AB - We tested the hypotheses that the glucagon response to hypoglycemia is reduced in patients who are approaching the insulin-deficient end of the spectrum of type 2 diabetes and that recent antecedent hypoglycemia shifts the glycemic thresholds for autonomic (including adrenomedullary epinephrine) and symptomatic responses to hypoglycemia to lower plasma glucose concentrations in type 2 diabetes. Hyperinsulinemic stepped hypoglycemic clamps (85, 75, 65, 55, and 45 mg/dl steps) were performed on two consecutive days, with an additional 2 h of hypoglycemia (50 mg/dl) in the afternoon of the first day, in 13 patients with type 2 diabetes --7 treated with oral hypoglycemic agents (OHA R(X); mean [+/- SD] HbA(1c) 8.6 +/ 1.1%) and 6 requiring therapy with insulin for an average of 5 years and with reduced C-peptide levels (insulin R(X), HbA(1c) 7.5 +/- 0.7%)---and 15 nondiabetic control subjects. The glucagon response to hypoglycemia was virtually absent (P = 0.0252) in the insulin-deficient type 2 diabetic patients (insulin R(X) mean [+/- SE] final values of 52 plus minus 16 vs. 93 plus minus 15 pg/ml in control subjects and 98 +/- 16 pg/ml in type 2 diabetic patients, OHA R(X) on day 1). Glucagon (P = 0.0015), epinephrine (P = 0.0002), and norepinephrine (P = 0.0138) responses and neurogenic (P = 0.0149) and neuroglycopenic (P = 0.0015) symptom responses to hypoglycemia were reduced on day 2 after hypoglycemia on day 1 in type 2 diabetic patients; these responses were not eliminated, but their glycemic thresholds were shifted to lower plasma glucose concentrations. In addition, the glycemic thresholds for these responses were at higher-than-normal plasma glucose concentrations (P = 0.0082, 0.0028, 0.0023, and 0.0182, respectively) at baseline (day 1) in OHA R(X) type 2 diabetic patients, with relatively poorly controlled diabetes. Because the glucagon response to falling plasma glucose levels is virtually absent and the glycemic thresholds for autonomic and symptomatic responses to hypoglycemia are shifted to lower glucose concentrations by recent antecedent hypoglycemia, patients with advanced type 2 diabetes, like those with type 1 diabetes, are at risk for hypoglycemia associated autonomic failure and the resultant vicious cycle of recurrent iatrogenic hypoglycemia. PMID- 11872674 TI - Systemic and local adrenergic regulation of muscle glucose utilization during hypoglycemia in healthy subjects. AB - Adrenergic responses are crucial for hypoglycemic recovery. Epinephrine increases glucose production, lipolysis, and peripheral insulin resistance as well as blood flow and glucose delivery. Sympathetic activation causes vasoconstriction and reduces glucose delivery. To determine the effects of alpha- and beta-adrenergic activity on muscle glucose uptake during hypoglycemia, we studied forearm blood flow (FBF) (plethysmography), arteriovenous glucose difference (AV-diff), and forearm glucose uptake (FGU) during insulin infusion with 60 min of euglycemia followed by 60 min of hypoglycemia. Twelve healthy subjects (27 plus minus 5 years of age) were randomized to intravenous propranolol (IV PROP, 80 microg/min), intravenous phentolamine (IV PHEN, 500 microg/min), intra-arterial propranolol (IA PROP, 25 microg/min), intra-arterial phentolamine (IA PHEN, 12 microg/min per 100 ml forearm tissue), and saline (SAL). FBF increased during hypoglycemia with SAL (P < 0.001) but not with IA or IV PROP. FGU (P = 0.015) and AV-diff (P = 0.099) fell during hypoglycemia with IA PROP but not with IV PROP. FBF increased during hypoglycemia with IA and IV PHEN (P < 0.005). AV-diff fell during hypoglycemia with IA and IV PHEN (P < 0.01), but FGU was unchanged. Blood pressure fell (P < 0.001), and adrenergic and neuroglycopenic symptoms increased with IV PHEN (P < 0.01). Thus, systemic but not local propranolol prevents a decrease in forearm glucose extraction during hypoglycemia, suggesting that epinephrine increases peripheral muscular insulin resistance through systemic effects. alpha-Adrenergic activation inhibits vasodilation and helps maintain brain glucose delivery. PMID- 11872675 TI - Increased insulin sensitivity in IGF-I receptor--deficient brown adipocytes. AB - Immortalized brown adipocyte cell lines have been generated from fetuses of mice deficient in the insulin-like growth factor I receptor gene (IGF-IR(-/-)), as well as from fetuses of wild-type mice (IGF-IR(+/+)). These cell lines maintained the expression of adipogenic- and thermogenic-differentiation markers and show a multilocular fat droplets phenotype. IGF-IR(-/-) brown adipocytes lacked IGF-IR protein expression; insulin receptor (IR) expression remained unchanged as compared with wild-type cells. Insulin-induced tyrosine autophosphorylation of the IR beta-chain was augmented in IGF-IR--deficient cells. Upon insulin stimulation, tyrosine phosphorylation of (insulin receptor substrate-1) IRS-1 was much higher in IGF-IR(-/-) brown adipocytes, although IRS-1 protein content was reduced. In contrast, tyrosine phosphorylation of IRS-2 decreased in IGF-IR- deficient cells; its protein content was unchanged as compared with wild-type cells. Downstream, the association IRS-1/growth factor receptor binding protein-2 (Grb-2) was augmented in the IGF-IR(-/-) brown adipocyte cell line. However, SHC expression and SHC tyrosine phosphorylation and its association with Grb-2 were unaltered in response to insulin in IGF-IR--deficient brown adipocytes. These cells also showed an enhanced activation of mitogen-activated protein kinase (MAPK) kinase (MEK1/2) and p42/p44 mitogen-activated protein kinase (MAPK) upon insulin stimulation. In addition, the lack of IGF-IR in brown adipocytes resulted in a higher mitogenic response (DNA synthesis, cell number, and proliferating cell nuclear antigen expression) to insulin than wild-type cells. Finally, cells lacking IGF-IR showed a much lower association between IR or IRS-1 and phosphotyrosine phosphatase 1B (PTP1B) and also a decreased PTP1B activity upon insulin stimulation. However, PTP1B/Grb-2 association remained unchanged in both cell types, regardless of insulin stimulation. Data presented here provide strong evidence that IGF-IR--deficient brown adipocytes show an increased insulin sensitivity via IRS-1/Grb-2/MAPK, resulting in an increased mitogenesis in response to insulin. PMID- 11872676 TI - Extreme insulin resistance of the central adipose depot in vivo. AB - Despite the well-described association between obesity and insulin resistance, the physiologic mechanisms that link these two states are poorly understood. The present study was performed to elucidate the role of visceral adipose tissue in whole-body glucose homeostasis. Dogs made abdominally obese with a moderately elevated fat diet had catheters placed into the superior mesenteric artery so that the visceral adipose bed could be insulinized discretely. Omental insulin infusion was extracted at approximately 27%, such that systemic insulin levels were lower than in control (portal vein) insulin infusions. Omental infusion did not lower systemic free fatty acid levels further than control infusion, likely because of the resistance of the omental adipose tissue to insulin suppression and the confounding lower systemic insulin levels. The arteriovenous difference technique showed that local infusion of insulin did suppress omental lipolysis, but only at extremely high insulin concentrations. The median effective dose for suppression of lipolysis was almost fourfold higher in the visceral adipose bed than for whole-body suppression of lipolysis. Thus, the omental adipose bed represents a highly insulin-resistant depot that drains directly into the portal vein. Increased free fatty acid flux to the liver may account for hepatic insulin resistance in the moderately obese state. PMID- 11872677 TI - Albumin binding of acylated insulin (NN304) does not deter action to stimulate glucose uptake. AB - NN304 [Lys(B29)-tetradecanoyl des(B30) human insulin] is a potentially therapeutic insulin analog designed to exhibit protracted glucose-lowering action. In dogs with infusion rates similar to insulin itself, NN304 exhibits similar glucose uptake (R(d)) stimulation with delayed onset of action. This compartmental modeling study was to determine if NN304 action could be accounted for by the approximately 2% unbound NN304 concentration. NN304 (or human insulin) (n = 6 each) was infused at 10.2 pmol center dot min(-1) center dot kg(-1) under euglycemic clamp conditions in anesthetized dogs. NN304 appearance in lymph, representing interstitial fluid (ISF), was slow compared with insulin (t(1/2) = 70 +/- 7 vs. 14 +/- 1 min, P < 0.001). R(d) was highly correlated with the ISF concentration for insulin and NN304 (r = 0.86 and 0.93, respectively), suggesting that slow transendothelial transport (TET) is responsible for sluggish NN304 action. Insulin and NN304 concentration data were fit to a two-compartment (plasma and ISF) model. NN304 plasma elimination and TET were reduced to 10 and 7% of insulin, respectively. Thus, there was reduction of NN304 transport, but not to the degree expected. In ISF, there was no reduction in NN304 elimination. Thus, this acylated insulin analog demonstrates blunted kinetics in plasma, and full efficacy in the compartment of action, ISF. PMID- 11872678 TI - Unprocessed proinsulin promotes cell survival during neurulation in the chick embryo. AB - We have chosen a vertebrate model accessible during neurulation, the chick, for analysis of endogenous insulin signaling and its contribution to early embryonic cell survival. Unlike rodents, humans and chickens have a single preproinsulin gene, facilitating its prepancreatic expression characterization. We show that in vivo interference with embryonic insulin signaling using antisense oligonucleotides against the insulin receptor increases apoptosis during neurulation. In contrast, high glucose administration does not increase the level of apoptosis in culture or in vivo. Exogenous insulin and, remarkably, proinsulin achieve similar survival protective effects at 10(-8) mol/l. The low abundant preproinsulin mRNA from the prepancreatic embryo is translated to a protein that remains as unprocessed proinsulin. This concurs with the absence of prohormone convertase 2 (PC2) in the embryo, whereas PC2 is present later in embryonic pancreas. A C-peptide--specific antibody stains proinsulin-containing neuroepithelial cells of the chick embryo in early neurulation, as well as other cells in mesoderm- and endoderm-derived structures in the 2.5-day embryo. We have determined by 5'-RACE (rapid amplification of cDNA ends), and confirmed by RNase protection assay, that prepancreatic and pancreatic proinsulin mRNA differ in their first exon, suggesting differential transcriptional regulation. All these data support the role of endogenous proinsulin in cell survival in the chick embryo during important pathophysiologic periods of early development. PMID- 11872679 TI - Attenuation of diabetic hyperphagia in neuropeptide Y--deficient mice. AB - The combined effects of increased hypothalamic signaling by neuropeptide Y (NPY) and decreased signaling by melanocortins are hypothesized to stimulate food intake when body fat stores are depleted. To investigate NPY's role in the hyperphagic response to uncontrolled diabetes, streptozotocin (STZ) (200 mg/kg intraperitoneally) or saline vehicle was given to NPY-deficient (Npy(--/--)) and wild-type (Npy(+/+)) mice. In Npy(+/+) mice, STZ-induced diabetes increased mean daily food intake to plateau values 50% above baseline intake (+2.0 +/- 0.6 g/day; P < or = 0.05), an effect that was not seen in STZ-treated Npy(--/--) mice (+0.8 +/- 0.1 g/day; NS), despite comparably elevated levels of plasma glucose and comparably decreased levels of body weight, fat content, and plasma leptin. Unlike the impaired feeding response to uncontrolled diabetes, Npy(--/--) mice exhibit intact hyperphagic responses to fasting (Erickson et al. [1], Nature 381:415-418, 1996). To investigate whether differences in hypothalamic melanocortin signaling can explain this discrepancy, we used in situ hybridization to compare the effects of STZ-diabetes and fasting on pro opiomelanocortin (POMC) and agouti-related peptide (AgRP) mRNA levels in the hypothalamic arcuate nucleus (ARC) of Npy(--/--) and Npy(+/+) mice. AgRP mRNA levels were increased by both fasting and STZ-diabetes, but the increase in STZ diabetes was small (50-80%) compared with the effect of fasting (approximately 20 fold increase of AgRP mRNA). STZ-diabetes also lowered POMC mRNA levels by 65% in the ARC of Npy(+/+) mice (P less-than-or-equal 0.05) but by only 11% in Npy(--/- ) mice (NS); fasting significantly lowered POMC mRNA levels in both genotypes. We conclude that NPY is required for both the increase of food intake and the decrease of hypothalamic POMC gene expression induced by uncontrolled diabetes. In contrast, NPY is not required for either of these responses when the stimulus is food deprivation. Moreover, fasting is a more potent stimulus to hypothalamic AgRP gene expression than is STZ-diabetes. Therefore, central nervous system melanocortin signaling appears to be suppressed more effectively by fasting than by uncontrolled diabetes, which provides a plausible explanation for differences in the feeding response to these two stimuli in mice lacking NPY. PMID- 11872680 TI - The fate of [U-(13)C]palmitate extracted by skeletal muscle in subjects with type 2 diabetes and control subjects. AB - The current study investigated the fate of a [U-(13)C]palmitate tracer extracted by forearm muscle in type 2 diabetic and control subjects. We studied seven healthy lean male subjects and seven obese male subjects with type 2 diabetes using the forearm muscle balance technique with continuous intravenous infusion of the stable isotope tracer [U-(13)C]palmitate under baseline conditions and during intravenous infusion of the nonselective beta-agonist isoprenaline (ISO; 20 ng *kg(-1) lean body mass* min(-1)). In skeletal muscle of control subjects, there was a significant release of (13)C-labeled oxidation products in the form of (13)CO(2) (15% of (13)C uptake from labeled palmitate) and a significant release of (13)C-labeled glutamine (release of (13)C-labeled atoms from glutamine was 6% of (13)C uptake from labeled palmitate), whereas in type 2 diabetic subjects there was no detectable release of (13)CO(2) and (13)C-glutamine, despite a significant uptake of [U-(13)C]palmitate (60% of control value). There was net uptake of arterial (13)C-labeled glutamate by forearm muscle in both groups. Also, the ISO-induced increase in arterial glutamine enrichment and arterial concentration of (13)C-glutamine was more pronounced in the diabetic group relative to control subjects. In view of the diminished ISO-induced release of (13)C-glutamine from type 2 diabetic muscle, the latter data indicate that more [U-(13)C]palmitate entered the liver in the diabetic group and was incorporated into newly synthesized glutamine and glutamate molecules. Thus, the lack of release of (13)C-labeled oxidation products by type 2 diabetic muscle during beta-adrenergic stimulation, despite significant [U-(13)C]palmitate uptake, indicates differences in the handling of fatty acids between type 2 diabetic subjects and healthy control subjects. PMID- 11872681 TI - Theophylline improves hypoglycemia unawareness in type 1 diabetes. AB - Iatrogenic hypoglycemias and the subsequent occurrence of hypoglycemia unawareness are well-known complications of intensive insulin therapy in type 1 diabetic patients that limit glycemic management. From a pharmacological point of view, the adenosine-receptor antagonist theophylline might be beneficial in the management of hypoglycemia unawareness. Theophylline stimulates the release of catecholamines and reduces cerebral blood flow, thereby facilitating stronger metabolic responses to and a prompter perception of decreasing glucose levels. To test the effect of theophylline on responses to hypoglycemia, we performed paired hyperinsulinemic-hypoglycemic clamp studies in 15 diabetic patients with hypoglycemia unawareness and 15 matched healthy control subjects. In random order, we concurrently infused either theophylline or placebo. Measurements included counterregulatory hormones, symptoms, hemodynamic parameters, and sweat detection using a dew-point electrode. Additionally, middle cerebral artery velocities (V(MCA)) using transcranial Doppler were monitored as an estimate of cerebral blood flow. When compared with placebo, theophylline significantly enhanced responses of plasma epinephrine, norepinephrine, and cortisol levels in both diabetic patients and control subjects. Because of the theophylline, sweat production started at approximately 0.3 mmol/l higher glucose levels in both groups (P < 0.01), and symptom scores in diabetic patients approached those in control subjects. Theophylline decreased V(MCA) in both groups (P < 0.001), but significantly greater in diabetic patients (P < 0.01), and prevented the hypoglycemia-induced increase of V(MCA) that occurred during the placebo studies. We conclude that theophylline improves counterregulatory responses to and perception of hypoglycemia in diabetic patients with impaired awareness of hypoglycemia. PMID- 11872683 TI - Association of fasting plasma glucose with heart rate recovery in healthy adults: a population-based study. AB - Diabetes is associated with abnormal autonomic function and increased mortality. Abnormal heart rate recovery after exercise, a measure of autonomic dysfunction, is also associated with increased mortality. The objective of this study was to determine the association of fasting plasma glucose with abnormal heart rate recovery and its prognostic importance in healthy adults. We studied 5,190 healthy adults who did not have medically treated diabetes (mean age 45 years, 39% women), were enrolled in the Lipid Research Clinics' Prevalence Study, and underwent exercise testing. Heart rate recovery was defined as the change from peak heart rate to that after 2 min of recovery; an abnormal value was < or = 42 bpm. All-cause mortality was assessed over 12 years. A total of 504 participants (10%) had impaired fasting glucose, and 131 (3%) had untreated diabetes. An abnormal heart rate recovery was found in 1,699 (33%). Compared with participants who had normal fasting plasma glucose, abnormal heart rate recovery was more common among those with impaired fasting glucose (42 vs. 31%; relative risk, 1.34; 95% confidence interval [CI], 1.20--1.50; P < 0.0001) and those with diabetes (50 vs. 31%; relative risk, 1.61; 95% CI, 1.35--1.92; P < 0.0001). Fasting plasma glucose remained an independent predictor of abnormal heart rate recovery even after adjustment for age, sex, and other confounders (P = 0.0003). An abnormal heart rate recovery added to impaired fasting plasma glucose for the prediction of death. Fasting plasma glucose is strongly and independently associated with abnormal heart rate recovery, even at nondiabetic levels. PMID- 11872684 TI - Effects of treatment with sulfonylurea drugs or insulin on ischemia-induced myocardial dysfunction in type 2 diabetes. AB - In patients with diabetes and coronary artery disease, the potential negative role of sulfonylurea drugs is under intensive investigation. We assessed the effects of treatment with glibenclamide or insulin on the extension of left ventricular myocardial dysfunction induced by acute ischemia. Nineteen consecutive patients with type 2 diabetes and coronary artery disease entered the study. Each patient was randomly assigned to either insulin or glibenclamide therapy. Treatment was crossed over after 12 weeks and maintained for another 12 weeks. At the end of each treatment, left ventricular myocardial function at rest and during dipyridamole infusion was studied by two-dimensional echocardiography under the same conditions of metabolic control. Glibenclamide or insulin treatment did not influence the rest values of left ventricular dimensions, left ventricular ejection fraction (LVEF), or wall motion score index (WMSI). Dipyridamole infusion, in patients receiving glibenclamide treatment, decreased LVEF (43 +/- 7 vs. 37 +/- 12%, P < 0.005) and increased WMSI (1.4 +/- 0.28 vs. 1.98 +/- 0.24, P < 0.001) compared with baseline values; during insulin treatment, LVEF (46 +/- 8 vs. 45 +/- 11%, NS) and WMSI (1.4 +/- 0.29 vs. 1.6 +/- 0.4, NS) did not change significantly. Peak stress LVEF was higher (45 +/- 11 vs. 37 +/- 12%, P < 0.001) and WMSI lower (1.6 +/- 0.4 vs. 1.98 +/- 0.24, P < 0.001) in patients receiving insulin. The results indicate that in patients with type 2 diabetes and coronary artery disease, ischemic myocardial dysfunction induced by dipyridamole infusion is less severe during treatment with insulin than with glibenclamide. Restitution of a preconditioning mechanism in insulin-treated patients may be the potential beneficial mechanism. PMID- 11872682 TI - The effects of rosiglitazone on insulin sensitivity, lipolysis, and hepatic and skeletal muscle triglyceride content in patients with type 2 diabetes. AB - We examined the effect of three months of rosiglitazone treatment (4 mg b.i.d.) on whole-body insulin sensitivity and in vivo peripheral adipocyte insulin sensitivity as assessed by glycerol release in microdialysis from subcutaneous fat during a two-step (20 and 120 mU.m(-2).min(-1)) hyperinsulinemic-euglycemic clamp in nine type 2 diabetic subjects. In addition, the effects of rosiglitazone on liver and muscle triglyceride content were assessed by (1)H-nuclear magnetic resonance spectroscopy. Rosiglitazone treatment resulted in a 68% (P < 0.002) and a 20% (P < 0.016) improvement in insulin-stimulated glucose metabolism during the low- and high- dosage-insulin clamps, respectively, which was associated with approximately 40% reductions in plasma fatty acid concentration (P < 0.05) and hepatic triglyceride content (P < 0.05). These changes were associated with a 39% increase in extramyocellular lipid content (P < 0.05) and a 52% increase in the sensitivity of peripheral adipocytes to the inhibitory effects of insulin on lipolysis (P = 0.04). In conclusion, these results support the hypothesis that thiazolidinediones enhance insulin sensitivity in patients with type 2 diabetes by promoting increased insulin sensitivity in peripheral adipocytes, which results in lower plasma fatty acid concentrations and a redistribution of intracellular lipid from insulin responsive organs into peripheral adipocytes. PMID- 11872685 TI - Coronary microvascular adaptation to myocardial metabolic demand can be restored by inhibition of iron-catalyzed formation of oxygen free radicals in type 2 diabetic patients. AB - Dilation of coronary vessels is impaired in diabetic patients when myocardial metabolic demand is increased. Deferoxamine (DFX) restores a normal dilation of epicardial coronary arteries. To assess the effects of DFX on metabolic coronary microvascular dilation in type 2 diabetic patients, coronary blood flow was measured using intracoronary Doppler and quantitative angiography in 17 type 2 diabetic patients with normal coronary arteries and without any other coronary risk factors. Measurements were made at baseline and during a cold pressor test (CPT), before and after intravenous administration of DFX. With a similar rate pressure product (RPP) increase during CPT before and after DFX (+21.1 +/- 8.7 vs. +20.5 +/- 8.9%, respectively), coronary blood flow increase was significantly enhanced after DFX (+31.8 +/- 16.7 vs. +6.3 +/- 12.9% before DFX, P < 0.001). Moreover, coronary resistance increased during CPT before DFX and decreased after DFX (+14.8 +/- 21.9 vs. -7.9 +/- 10.9%, respectively, P < 0.001). Lastly, the negative relationship between coronary blood flow and RPP before DFX (R = 0.546, P < 0.05) was changed in a positive relationship after DFX (R = 0.518, P < 0.05). In conclusion, in type 2 diabetic patients, inhibition of iron-catalyzed oxidative reactions by DFX restored dilation of the coronary microcirculation and a normal matching between myocardial metabolic demand and coronary blood flow. PMID- 11872686 TI - Morphometry of dorsal root ganglion in chronic experimental diabetic neuropathy. AB - Chronic hyperglycemia results in a predominantly sensory neuropathy. Recent studies suggest that dorsal root ganglion (DRG) neurons comprise a specific target and may be responsible for the important complication of diabetic sensory neuropathy, since hyperglycemia for longer than 6 months results in a vacuolar ganglionopathy with associated radiculopathy and distal sensory neuropathy. We undertook morphometric analysis of L5 DRG neurons in seven diabetic rats and six age- and sex-matched littermates. Nerve conduction studies were also performed, and neuropathy was confirmed. Diabetes was induced with streptozotocin; duration of diabetes was 12 months. The DRG count for control rats was 15,304 +/- 991 neurons. Two of seven diabetic DRG counts were reduced, but the group mean count at 14,847 plus minus 1,524 was not significantly reduced. The number of small neurons (type B) considerably exceeded that of large neurons (type A), at a ratio of 71:29. The percentage of large cells was significantly reduced in diabetic compared with control rats (P = 0.01). The large-diameter population can be subdivided into two groups; with this subdivision, the number of neurons < 50 microm was not reduced in samples from diabetic rats, but the neurons of largest size (> or = 50 microm) were significantly reduced (by 41%). PMID- 11872687 TI - Deconstructing and reconstructing obesity-induced diabetes (diabesity) in mice. AB - Obesity-driven type 2 diabetes (diabesity) involves complex genetic and environmental interactions to trigger disease. Here, we combine variable numbers of known quantitative trait loci (QTL) for obesity and diabetes contributed by New Zealand Obese (NZO/HlLt) and Nonobese Nondiabetic (NON/Lt) strains in the form of 10 interval-directed recombinant congenic strains (RCS), with NON/Lt as the background strain, to dissect the genetic interactions involved. All 10 RCS gain significantly more weight than the NON parental strain, but none are as obese as the parental, diabetes-prone NZO. Diabetes development in these RCS at F12 ranges between 0 and 100%, depending on genetic constitution. RCS-2, -1, and 10 represent a step-wise increase in numbers of specific diabetogenic QTL, resulting in a step-wise increase in diabetes incidence. RCS-10 recreates the 100% incidence seen in (NZOxNON)F1 males, but with less weight gain. Similarly, RCS-6, -7, -8, and -9 represent diabetes-prone strains with different combinations of diabetogenic QTL. RCS-3, -4, and -5 represent obese strains that do not transit to diabetes. Because these obesity and diabetes syndromes reflect different collections of QTL, rather than null mutations in the leptin or leptin receptor genes, they are extremely relevant as models for the polygenic obesity/diabesity syndromes in humans. PMID- 11872688 TI - A genome-wide scan for loci linked to plasma levels of glucose and HbA(1c) in a community-based sample of Caucasian pedigrees: The Framingham Offspring Study. AB - Elevated blood glucose levels are the hallmark of type 2 diabetes as well as a powerful risk factor for development of the disease. We conducted a genome-wide search for diabetes-related genes, using measures of glycemia as quantitative traits in 330 pedigrees from the Framingham Heart Study. Of 3,799 attendees at the 5th Offspring Study exam cycle (1991--1995), 1,461, 1,251, and 771 men (49%) and women provided information on levels of 20-year mean fasting glucose, current fasting glucose, and HbA(1c), respectively, and 1,308 contributed genotype data (using 401 microsatellite markers with an average spacing of 10 cM). Levels of glycemic traits were adjusted for age, cigarette smoking, alcohol and estrogen use, physical activity, and BMI. We ranked standardized residuals from these models, created normalized deviates from the ranks, and used the variance component model implemented in SOLAR (Sequential Oligogenic Linkage Analysis Routines) to evaluate linkage to normalized deviates as quantitative traits. We found peak evidence for linkage to 20-year mean fasting glucose levels on chromosome 1 at approximately 247 cM from p-telomere (pter) (multipoint logarithm of odds [LOD] 2.33) and on chromosome 10 at approximately 86 cM from pter (multipoint LOD 2.07); to current fasting glucose levels on chromosome 1 at approximately 218 cM from pter (multipoint LOD 1.80) and on chromosome 10 at approximately 96 cM from pter (multipoint LOD 2.15); and to HbA(1c) levels on chromosome 1 at approximately 187 cM (multipoint LOD 2.81). This analysis of unselected European Caucasian pedigrees suggests localization of quantitative trait loci influencing glucose homeostasis on chromosomes 1q and 10q. Findings at approximately 187--218 cM on chromosome 1 appear to replicate linkage reported in previous studies of other populations, pointing to this large chromosomal region as worthy of more detailed scrutiny in the search for type 2 diabetes susceptibility genes. PMID- 11872689 TI - Factors of insulin resistance syndrome--related phenotypes are linked to genetic locations on chromosomes 6 and 7 in nondiabetic mexican-americans. AB - Insulin resistance syndrome (IRS)-related phenotypes, such as hyperinsulinemia, obesity-related traits, impaired glucose tolerance, dyslipidemia, and hypertension, tend to cluster into factors. We attempted to identify loci influencing the factors of IRS-related phenotypes using phenotypic data from 261 nondiabetic subjects distributed across 27 low-income Mexican-American extended families. Principal component factor analyses were performed using eight IRS related phenotypes: fasting glucose (FG), fasting specific insulin (FSI), BMI, systolic blood pressure (SBP), diastolic blood pressure (DBP), HDL cholesterol, ln triglycerides (ln TGs), and leptin (LEP). The factor analysis yielded three factors: factor 1 (BMI, LEP, and FSI), factor 2 (DBP and SBP), and factor 3 (HDL and ln TG). We conducted multipoint variance components linkage analyses on these factors with the program SOLAR using a 10--15 cM map. We found significant evidence for linkage of factor 1 to two regions on chromosome 6 near markers D6S403 (logarithm of odds [LOD] = 4.2) and D6S264 (LOD = 4.9). We also found strong evidence for linkage of factor 3 to a genetic location on chromosome 7 between markers D7S479 and D7S471 (LOD = 3.2). In conclusion, we found substantial evidence for susceptibility loci on chromosomes 6 and 7 that appear to influence the factors representing the IRS-related phenotypes in Mexican Americans. PMID- 11872690 TI - A genomewide linkage scan for abdominal subcutaneous and visceral fat in black and white families: The HERITAGE Family Study. AB - Abdominal visceral fat (AVF), abdominal subcutaneous fat (ASF), and abdominal total fat (ATF) were measured using a computed tomography scan, both before (baseline) and after (post) a 20-week endurance exercise training protocol in the HERITAGE Family Study. Each of the baseline and response (post minus baseline) measures was adjusted for several covariates, including total fat mass, and responses to training were further adjusted for baseline levels. Multipoint variance components linkage analysis using a genomewide scan of 344 markers was conducted separately by race using race-specific allele frequencies. Several promising results (P < 0.0023) were obtained. For baseline AVF, the best evidence was on 2q22.1 and 2q33.2-q36.3 (including the IRS1 locus) in whites, with suggestive findings on 7q22.2-q31.3 (including the LEP locus) in blacks. Although several regions were indicated for baseline ASF, only 4q31.22-q32.2 and 11p15.4 p11.2 replicated the results of another study. For responses to training, promising results were limited to ASF and ATF primarily on 7q36.2 (including NOS3) in blacks, with suggestive regions (P < 0.01) on 1q21.2-q24.1 (S100A, ATP1A2, and ATP1B1), 10q25.2 (ADRA2A), and 11p15.5 (IGF2). In summary, the 4q and 11p regions have now been implicated in two independent studies for ASF; further research is warranted to identify the genes and mutations in these regions that are responsible for fat accumulation in the abdominal depot. Additional work in an independent sample is needed to verify the linkages for baseline AVF as well as the response measures. PMID- 11872691 TI - A genome-wide search for linkage-disequilibrium with type 1 diabetes in a recent genetically isolated population from the Netherlands. AB - Type 1 diabetes has a substantial genetic component, with consistent evidence for a susceptibility locus in the HLA-DR/DQ region (chromosome 6p) and the insulin gene region (chromosome 11p). Genome scans have identified >18 other genomic regions that may harbor putative type 1 diabetes genes. However, evidence for most regions varies in different data sets. Given the genetic heterogeneity of type 1 diabetes, studies in homogeneous genetically isolated populations may be more successful in mapping susceptibility loci than in complex outbred populations. We describe a genome-wide search in a recently Dutch isolated population. We identified 43 patients that could be traced back to a common ancestor within 15 generations and performed a genome-wide scan using a combined linkage- and association-based approach. In addition to the HLA locus, evidence for type 1 diabetes loci was observed on chromosome 8q24 (marker D8S1128) and on chromosome 17q24 (marker D17S2059). Both the 8q and 17q localization are supported by allele-sharing at adjacent markers in affected individuals. Statistical evidence for a conserved ancestral haplotype was found for chromosome 8q24. PMID- 11872692 TI - Human resistin gene, obesity, and type 2 diabetes: mutation analysis and population study. AB - The hormone resistin has been suggested to link obesity to type 2 diabetes by modulating steps in the insulin-signaling pathway and inducing insulin resistance. Thus, the resistin gene represents a potential candidate for the etiology of insulin resistance and type 2 diabetes. In this study, we analyzed the coding sequence of the three exons of the resistin gene, together with its 5' regulatory region and 3' untranslated region (UTR), by single-strand conformation polymorphism (SSCP) in 58 type 2 diabetic subjects, 59 obese subjects, and 60 normal subjects. Only one sequence variant was detected in the resistin gene. Sequencing of this variant revealed the presence of a single nucleotide substitution (SNP) in the 3'-UTR of exon 3 (G1326A) [corrected]. Because 3'-UTR SNPs have been shown to affect gene expression, we examined the frequency of this SNP in 591 subjects (198 obese subjects, 207 diabetic subjects, and 186 control subjects) by PCR amplification and BseRI digestion. No significant association was found between the G1326A [corrected] variant and diabetes and obesity. Comparison of clinical and metabolic parameters between G1326A [corrected] carriers and noncarriers again showed no significant difference. In conclusion, our data suggest that genetic defects of the resistin gene are unlikely to play a role in the etiology of these common disorders in our population. PMID- 11872693 TI - Systematic search for single nucleotide polymorphisms in the resistin gene: the absence of evidence for the association of three identified single nucleotide polymorphisms with Japanese type 2 diabetes. AB - Resistin is a novel polypeptide specifically secreted from adipocytes, and its serum levels are increased in obese diabetic mice. Resistin antagonizes insulin and could account for insulin resistance. To determine whether there are single nucleotide polymorphisms (SNPs) in the resistin gene associated with type 2 diabetes, sequences for 24 Japanese type 2 diabetic patients were initially analyzed using PCR direct sequencing. Three SNPs were found in the introns, but none were present in the coding regions. The allele frequencies of genomic 167C>T, +157C>T, and +299G>A in 99 Japanese control subjects were determined to be 3.5, 6.6, and 39.4%, respectively. In each pair of these SNPs, linkage disequilibria were found between either -167C>T and +299G>A or +157C>T and +299G>A. A linkage disequilibrium was also detected among -167C>T, +157C>T, and +299G>A, and only four of the eight possible haplotypes defined by these SNPs were found. A comparison of the frequencies of these SNPs and haplotypes between 99 type 2 diabetes and 99 control subjects revealed no evidence for any association. These identified SNPs, which were in linkage disequilibrium, represent potentially useful tools for searching for their association with specific phenotypes of diabetes. PMID- 11872694 TI - The proline 12 alanine substitution in the peroxisome proliferator--activated receptor-gamma2 gene is associated with lower lipoprotein lipase activity in vivo. AB - Lipoprotein lipase (LPL) plays a key role in lipid metabolism by hydrolyzing triglycerides in circulating lipoproteins. Low LPL activity has been linked to coronary artery disease (CAD), but the factors influencing LPL expression are not completely understood. Peroxisome proliferator--activated receptor (PPAR)-gamma is a nuclear receptor regulating lipid and glucose metabolism, and a PPAR responsive element is present in the LPL promoter. We determined the Pro12Ala polymorphism in the PPAR-gamma2 gene in 194 male CAD patients because this allele is associated with decreased PPAR activity and reduced LPL promoter activity in vitro. Presence of 12Ala was associated with 20% lower LPL activity in postheparin plasma (141 +/- 58 vs. 177 +/- 77 nmol.ml(-1).min(-1), P < 0.005). Remarkably, the influence of 12Ala on LPL was greater than that of the frequent polymorphisms (HindIII +9%, PvuII +/- 0%, 447stop +12%) in the LPL gene itself. To confirm these results in a different group of patients, we analyzed 100 diabetic patients in whom the 12Ala allele was also associated with lower LPL activity (12Ala: 132 +/- 88 vs. 190 +/- 129 nmol.ml(-1).min(-1), P < 0.05). Our data demonstrate that the Pro12Ala substitution in PPAR-gamma2 is associated with lower LPL activity in vivo and provides a new target for the analysis of genetic influences on LPL activity and CAD risk. PMID- 11872695 TI - Relationship between TaqIB cholesteryl ester transfer protein gene polymorphism and macrovascular complications in Japanese patients with type 2 diabetes. AB - Cholesteryl ester transfer protein (CETP) is a key regulating factor of lipid metabolism, and the polymorphism of its gene may therefore be a candidate for modulating the lipid parameters, altering the susceptibility to atherosclerosis in type 2 diabetic subjects. In a group of 443 unrelated Japanese patients with type 2 diabetes, we studied the B1B2 polymorphism at the CETP locus, which is detectable with the restriction enzyme TaqI. Patients were separated into three groups according to genotype and compared based on their clinical characteristics, lipid parameters, and macrovascular complications. The B2 allele was associated in a dose-dependent fashion with higher HDL cholesterol and apolipoprotein AI levels, together with lower CETP concentrations. Furthermore, the prevalence of macrovascular complications, such as coronary heart disease, arteriosclerosis obliterans, and cerebral vascular disease, was significantly higher in subjects with the B1B1 genotype. Multiple logistic regression analysis also showed that the B1 allele of CETP genotype was associated with the incidence of these three complications independently of other risk factors. Thus, in type 2 diabetic patients, the B1B2 polymorphism of CETP gene is likely to be a strong genetic predictor of macrovascular complications. PMID- 11872696 TI - K(IR)6.2 polymorphism predisposes to type 2 diabetes by inducing overactivity of pancreatic beta-cell ATP-sensitive K(+) channels. AB - E23K, a common single nucleotide polymorphism in K(IR)6.2, the pore-forming subunit of pancreatic beta-cell ATP-sensitive K(+) channels, significantly enhanced open probability of these channels, thus reducing their sensitivity toward inhibitory ATP(4-) and increasing the threshold concentration for insulin release. Previous association studies and high allelic frequency suggest this effect to critically inhibit secretion and play a major role in pathogenesis of common type 2 diabetes. Based on evidence for functional relevance of E23K in both the heterozygous (E/K; with E in position 23 of K(IR)6.2 in one allele and K in the other) and homozygous (K/K; with K in position 23 of K(IR)6.2 in both alleles) genotype, we propose a model in which enhanced susceptibility to type 2 diabetes is associated with evolutionary advantage of the E/K state. PMID- 11872697 TI - Association between a novel variant of the human type 2 deiodinase gene Thr92Ala and insulin resistance: evidence of interaction with the Trp64Arg variant of the beta-3-adrenergic receptor. AB - Thyroid hormone action is an important determinant of energy and glucose metabolism. T4 metabolism is regulated by the deiodinases of which type 2 is expressed in humans in skeletal muscle and brown adipose tissue, where its transcription is stimulated by the beta-3 adrenergic pathway. We performed molecular scanning of the human type 2 deiodinase (DIO2) gene and evaluated a novel variant for associations with obesity and insulin resistance, assessing both the main effect and interaction with the Trp64Arg beta-3--adrenergic receptor (ADRB3) variant. Molecular scanning of DIO2 in 50 obese Caucasians demonstrated a Thr92Ala variant. Association studies in 972 nondiabetic patients, 135 of whom underwent euglycemic-hyperinsulinemic clamps, showed that subjects with the Thr92Ala variant had lower glucose disposal rate (0.54 plus minus 0.02 mg center dot min(-1) center dot kg(-1) fat-free mass Ala92 homozygotes vs. 0.44 plus minus 0.02 Ala92 heterozygotes vs. 0.42 plus minus 0.04 Thr92 homozygotes, P = 0.0088). Association analysis of the entire group showed significant evidence for a synergistic effect between the Thr92Ala DIO2 and Trp64Arg ADRB3 variants on BMI (both variants 34.3 plus minus 0.9 kg/m(2) vs. neither variant 33.1 plus minus 0.4 kg/m(2), P = 0.04 for interaction). To our knowledge, Thr92Ala is the first description of a missense mutation of DIO2. This variant strongly associates with insulin resistance and, in subjects with the Trp64Arg ADRB3 variant, an increased BMI, suggesting an interaction between these two common gene variants. PMID- 11872698 TI - Variations in insulin secretion in carriers of gene variants in IRS-1 and -2. AB - Associations between type 2 diabetes (and/or parameters contributing to glucose homeostasis) and genetic variation in the genes encoding insulin receptor substrate (IRS)-1 and -2 have been reported in several populations. Recently, it has been reported that the Gly(972)Arg variant in IRS-1 was associated with reduced insulin secretion during hyperglycemic clamps in German subjects with normal glucose tolerance. We have examined glucose-stimulated insulin secretion in relation to gene variants in the IRS-1 (Gly(972)Arg) and IRS-2 (Gly(1057)Asp) genes in two Dutch cohorts. Subjects with normal (n = 64) or impaired (n = 94) glucose tolerance underwent 3-h hyperglycemic clamps at 10 mmol/l glucose. All subjects were genotyped for the IRS-1 and IRS-2 variants by PCR-RFLP--based methods. We did not observe any significant difference in both first- and second phase insulin secretion between carriers and noncarriers of both gene variants, nor was there evidence for an association with other diabetes-related parameters. We conclude that the common gene variants in IRS-1 and IRS-2 are not associated with altered glucose-stimulated insulin secretion in two populations from the Netherlands. PMID- 11872699 TI - Status of research funded by the American Diabetes Association: year 3. PMID- 11872701 TI - Identification and functional characterization of flgM, a gene encoding the anti sigma 28 factor in Pseudomonas aeruginosa. AB - We describe here the functional characterization of the putative flgM gene of Pseudomonas aeruginosa. FlgM of P. aeruginosa is most similar to FlgM of Vibrio parahaemolyticus. A conserved region is present in the C-terminal half of the FlgM of P. aeruginosa and in FlgM homologues of other organisms that includes the sigma(28) binding domain. A role for the flgM gene of P. aeruginosa in motility was demonstrated by its inactivation. The beta-galactosidase activity of a transcriptional fusion of the fliC promoter to lacZ was upregulated in the flgM mutant, suggesting that the activity of FliA, the sigma factor that regulates fliC, was increased. Consistent with these results, an increased amount of flagellin was demonstrated in the flgM mutant of P. aeruginosa strain PAK by Western blot, suggesting that FlgM negatively regulates transcription of fliC by inhibiting the activity of FliA. Direct interaction of the P. aeruginosa FlgM with the alternative sigma factor sigma(28) was demonstrated by utilizing the yeast two-hybrid system. Three putative consensus sigma(54) recognition sites and one sigma(28) site were found in the flgM upstream region. However, analysis of the transcriptional fusion of the flgM promoter to lacZ in different mutant backgrounds showed that the flgM promoter was not entirely dependent on either sigma(28) or sigma(54). A transcript was detected by primer extension that was 8 bp downstream of the consensus sigma(28)-binding site. Thus, a system for the control of flagellin synthesis by FlgM exists in P. aeruginosa that is different from that in the enteric bacteria and seems to be most similar to that of V. cholerae where both sigma(28)-dependent and -independent mechanisms of transcription exist. PMID- 11872700 TI - Mutational analysis of the TonB1 energy coupler of Pseudomonas aeruginosa. AB - Siderophore-mediated iron transport in Pseudomonas aeruginosa is dependent upon the cytoplasmic membrane-associated TonB1 energy coupling protein for activity. To assess the functional significance of the various regions of this molecule and to identify functionally important residues, the tonB1 gene was subjected to site directed mutagenesis, and the influence on iron acquisition was determined. The novel N-terminal extension of TonB1, which is absent in all other examples of TonB, was required for TonB1 activity in both P. aeruginosa and Escherichia coli. Appending it to the N terminus of the nonfunctional (in P. aeruginosa) Escherichia coli TonB protein (TonB(Ec)) rendered TonB(Ec) weakly active in P. aeruginosa and did not compromise the activity of this protein in E. coli. Elimination of the membrane-spanning, presumed membrane anchor sequence of TonB1 abrogated TonB1 activity in P. aeruginosa and E. coli. Interestingly, however, a conserved His residue within the membrane anchor sequence, shown to be required for TonB(Ec) function in E. coli, was shown here to be essential for TonB1 activity in E. coli but not in P. aeruginosa. Several mutations within the C terminal end of TonB1, within a region exhibiting the greatest similarity to other TonB proteins, compromised a TonB1 contribution to iron acquisition in both P. aeruginosa and E. coli, including substitutions at Tyr264, Glu274, Lys278, and Asp304. Mutations at Pro265, Gln293, and Val294 also impacted negatively on TonB1 function in E. coli but not in P. aeruginosa. The Asp304 mutation was suppressed by a second mutation at Glu274 of TonB1 but only in P. aeruginosa. Several TonB1 TonB(Ec) chimeras were constructed, and assessment of their activities revealed that substitutions at the N or C terminus of TonB1 compromised its activity in P. aeruginosa, although chimeras possessing an E. coli C terminus were active in E. coli. PMID- 11872702 TI - The global regulatory hns gene negatively affects adhesion to solid surfaces by anaerobically grown Escherichia coli by modulating expression of flagellar genes and lipopolysaccharide production. AB - The initial binding of bacterial cells to a solid surface is a critical and essential step in biofilm formation. In this report we show that stationary-phase cultures of Escherichia coli W3100 (a K-12 strain) can efficiently attach to sand columns when they are grown in Luria broth medium at 28 degrees C in fully aerobic conditions. In contrast, growth in oxygen-limited conditions results in a sharp decrease in adhesion to hydrophilic substrates. We show that the production of lipopolysaccharide (LPS) and of flagella, as well as the transcription of the fliC gene, encoding the major flagellar subunit, increases under oxygen-limited conditions. Inactivation of the global regulatory hns gene counteracts increased production of LPS and flagella in response to anoxia and allows E. coli W3100 to attach to sand columns even when it is grown under oxygen-limited conditions. We propose that increased production of the FliC protein and of LPS in response to oxygen limitation results in the loss of the ability of E. coli W3100 to adhere to hydrophilic surfaces. Indeed, overexpression of the fliC gene results in a decreased adhesion to sand even when W3100 is grown in fully aerobic conditions. Our observations strongly suggest that anoxia is a negative environmental signal for adhesion in E. coli. PMID- 11872703 TI - Bordetella interspecies allelic variation in AlcR inducer requirements: identification of a critical determinant of AlcR inducer responsiveness and construction of an alcR(Con) mutant allele. AB - Previous studies established the critical roles of AlcR and alcaligin inducer in positive regulation of alcaligin siderophore biosynthesis and transport genes in Bordetella pertussis and Bordetella bronchiseptica. Transcriptional analyses using plasmid-borne alcR genes of B. pertussis UT25 and B. bronchiseptica B013N to complement the alcR defect of B. bronchiseptica strain BRM13 (Delta alcR1 alcA::mini-Tn5 lacZ1) revealed interspecies differences in AlcR inducer requirements for activation of alcABCDER operon transcription. Whereas the B. pertussis UT25 AlcR protein retained strong inducer dependence when produced from multicopy plasmids, B. bronchiseptica B013N alcR partially suppressed the alcaligin requirement for transcriptional activation. Functional analysis of AlcR chimeras produced by interspecies domain swapping and interspecies reciprocal site-specific mutagenesis determined that the phenotypic difference in AlcR inducer dependence was due to a single amino acid difference within the proposed inducer-binding and multimerization domain of AlcR. Structural predictions guided the design of a mutant AlcR protein with a single amino acid substitution at this critical position, AlcR(S103T), that was fully constitutive not only when produced from multicopy plasmids but also at a single-copy gene dosage. These results indicate that AlcR residue 103 affects a critical determinant of alcaligin inducer dependence of AlcR-mediated transcriptional activation. The alcR(S103T) mutant allele is the first alcR(Con) mutant allele identified. PMID- 11872704 TI - The DevT protein stimulates synthesis of FruA, a signal transduction protein required for fruiting body morphogenesis in Myxococcus xanthus. AB - Fruiting body formation in Myxococcus xanthus involves three morphologic stages-- rippling, aggregation, and sporulation---all of which are induced by the cell surface-associated C-signal. We analyzed the function of the DevT protein, a novel component in the C-signal response pathway. A mutant carrying an in-frame deletion in the devT gene displays delayed aggregation and a cell autonomous sporulation defect, whereas it remains rippling proficient. To further define the function of DevT, the methylation pattern of FrzCD, a cytoplasmic methyl accepting chemotaxis protein homologue, was examined in the Delta devT mutant, and we found that DevT is required for methylation of FrzCD during development. Specifically, DevT was found to be required for the C-signal-dependent methylation of FrzCD. The Delta devT mutant produced wild-type levels of C signal. However, accumulation of the FruA response regulator protein, which is essential for the execution of the C-signal-dependent responses, was reduced in the Delta devT mutant. The DevT protein was found to stimulate the developmentally activated transcription of the fruA gene. Epistasis analyses indicate that DevT acts independently of the A- and E-signals to stimulate fruA transcription. These findings suggest that the developmental defects of the Delta devT mutant are associated with a lack of FruA to ensure a proper response to the C-signal during the aggregation and sporulation stages of development. PMID- 11872705 TI - Salicylate 5-hydroxylase from Ralstonia sp. strain U2: a monooxygenase with close relationships to and shared electron transport proteins with naphthalene dioxygenase. AB - The genes from the oxygenase cluster nagAaGHAbAcAd of naphthalene-degrading Ralstonia sp. strain U2 were cloned and overexpressed. Salicylate 5-hydroxylase (S5H) activity, converting salicylate to gentisate, was present in vitro only in the single extract of cells with overexpressed nagAaGHAb or in a mixture of three cell extracts containing, respectively, NagGH (the oxygenase components), NagAa (ferredoxin reductase), and NagAb (ferredoxin). Each of the three extracts required for S5H activity was rate limiting in the presence of excess of the others but, when in excess, did not affect the rate of catalysis. S5H catalyzed the 5-hydroxylation of the aromatic rings of 3- and 4-substituted salicylates. However, the methyl group of 5-methylsalicylate was hydroxylated to produce the 5 hydroxymethyl derivative and the 6-position on the ring of 5-chlorosalicylate was hydroxylated, producing 5-chloro-2,6-dihydroxybenzoate. In an assay for the nag naphthalene dioxygenase (NDO) based on the indole-linked oxidation of NADH, three extracts were essential for activity (NagAcAd, NagAa, and NagAb). NDO and S5H were assayed in the presence of all possible combinations of the nag proteins and the corresponding nah NDO proteins from the "classical" naphthalene degrader P. putida NCIMB9816. All three oxygenase components functioned with mixed combinations of the electron transport proteins from either strain. The S5H from strain U2 is a unique monooxygenase which shares sequence similarity with dioxygenases such as NDO but is also sufficiently similar in structure to interact with the same electron transport chain and probably does so in vivo during naphthalene catabolism in strain U2. PMID- 11872706 TI - Involvement of superoxide dismutases in the response of Escherichia coli to selenium oxides. AB - Selenium can provoke contrasting effects on living organisms. It is an essential trace element, and low concentrations have beneficial effects, such as the reduction of the incidence of cancer. However, higher concentrations of selenium salts can be toxic and mutagenic. The bases for both toxicity and protection are not clearly understood. To provide insights into these mechanisms, we analyzed the proteomic response of Escherichia coli cells to selenate and selenite treatment under aerobic conditions. We identified 23 proteins induced by both oxides and ca. 20 proteins specifically induced by each oxide. A striking result was the selenite induction of 8 enzymes with antioxidant properties, particularly the manganese and iron superoxide dismutases (SodA and SodB). The selenium inductions of sodA and sodB were controlled by the transcriptional regulators SoxRS and Fur, respectively. Strains with decreased superoxide dismutase activities were severely impaired in selenium oxide tolerance. Pretreatment with a sublethal selenite concentration triggered an adaptive response dependent upon SoxRS, conferring increased selenite tolerance. Altogether, our data indicate that superoxide dismutase activity is essential for the cellular defense against selenium salts, suggesting that superoxide production is a major mechanism of selenium toxicity under aerobic conditions. PMID- 11872708 TI - Identification of an acetoacetyl coenzyme A synthetase-dependent pathway for utilization of L-(+)-3-hydroxybutyrate in Sinorhizobium meliloti. AB - D-(-)-3-Hydroxybutyrate (DHB), the immediate depolymerization product of the intracellular carbon store poly-3-hydroxybutyrate (PHB), is oxidized by the enzyme 3-hydroxybutyrate dehydrogenase to acetoacetate (AA) in the PHB degradation pathway. Externally supplied DHB can serve as a sole source of carbon and energy to support the growth of Sinorhizobium meliloti. In contrast, wild type S. meliloti is not able to utilize the L-(+) isomer of 3-hydroxybutyrate (LHB) as a sole source of carbon and energy. In this study, we show that overexpression of the S. meliloti acsA2 gene, encoding acetoacetyl coenzyme A (acetoacetyl-CoA) synthetase, confers LHB utilization ability, and this is accompanied by novel LHB-CoA synthetase activity. Kinetics studies with the purified AcsA2 protein confirmed its ability to utilize both AA and LHB as substrates and showed that the affinity of the enzyme for LHB was clearly lower than that for AA. These results thus provide direct evidence for the LHB-CoA synthetase activity of the AcsA2 protein and demonstrate that the LHB utilization pathway in S. meliloti is AcsA2 dependent. PMID- 11872709 TI - Rhodospirillum rubrum possesses a variant of the bchP gene, encoding geranylgeranyl-bacteriopheophytin reductase. AB - The bchP gene product of Rhodobacter sphaeroides is responsible for the reduction of the isoprenoid moiety of bacteriochlorophyll (Bchl) from geranylgeraniol (GG) to phytol; here, we show that this enzyme also catalyzes the reduction of the isoprenoid moiety of bacteriopheophytin (Bphe). In contrast, we demonstrate that a newly identified homolog of this gene in Rhodospirillum rubrum encodes an enzyme, GG-Bphe reductase, capable of reducing the isoprenoid moiety of Bphe only. We propose that Rhodospirillum rubrum is a naturally occurring bchP mutant and that an insertion mutation may have been the initial cause of a partial loss of function. Normal BchP function can be restored to Rhodospirillum rubrum, creating a new transconjugant strain possessing Bchl esterified with phytol. We speculate on the requirement of Rhodospirillum rubrum for phytylated Bphe and on a potential link between the absence of LH2 and of phytylated Bchl from the wild type bacterium. The identification of a second role for the fully functional BchP in catalyzing the synthesis of phytylated Bphe strongly suggests that homologs of this enzyme may be similarly responsible for the synthesis of phytylated pheophytin in organisms possessing photosystem 2. In addition to bchP, other members of a photosynthesis gene cluster were identified in Rhodospirillum rubrum, including a bchG gene, demonstrated to encode a functional Bchl synthetase by complementation of a Rhodobacter sphaeroides mutant. PMID- 11872707 TI - The histone-like protein HU does not obstruct movement of T7 RNA polymerase in Escherichia coli cells but stimulates its activity. AB - In vivo, RNA polymerases (RNAPs) do not transcribe naked DNA but do transcribe protein-associated DNA. Studies with the model enzyme T7 RNAP have shown that, in eukaryotic cells or in vitro, nucleosomes can inhibit both transcription initiation and elongation. We examine here whether the presence of HU, one of the major histone-like proteins in Escherichia coli cells (the genuine milieu for T7 RNAP) affects its activity. An engineered lac operon fused to the T7 late promoter was introduced into the chromosome of T7 RNAP-producing strains that either overexpress HU or lack it. The flows of RNAP that enter and exit this operon were compared with regard to the content of HU. We found that the fraction of T7 RNAP molecules that do not reach the end of the lac operon (ca. 15%) is the same whether the host cells overexpressed HU or lacked it: thus, the enzyme either freely displaces HU or transcribes through it. However, in these cells, the transcript yield was increased when HU is overexpressed and decreased in the hup mutants, presumably reflecting changes in DNA supercoiling. Thus, in contrast to eukaryotic nucleosomes, HU does not impair T7 RNAP activity but has a stimulatory effect. Finally, our results suggest that HU can also influence mRNA stability in vivo. PMID- 11872710 TI - Phenotypic selection and phase variation occur during alfalfa root colonization by Pseudomonas fluorescens F113. AB - During colonization of the alfalfa rhizosphere, Pseudomonas fluorescens F113 undergoes phenotypic variation, resulting in the appearance of colonies with different morphology. Among phenotypic variants, three isolates, C, F, and S were selected, with the C variant showing colony morphology identical to that of the inoculated wild-type strain and F and S having a translucent and diffuse morphology. Phenotypic variants F and S were shown to preferentially colonize distal parts of the roots and showed alterations in motility, swimming faster than the C variant and swarming under conditions that did not allow swarming of the C variant. The motility behavior correlated with overproduction of the fliC encoded protein flagellin but not with hyperflagellation. Flagella of the F and S variants were several times longer than those of the C variant, and overproduction of flagellin was regulated at the transcriptional level. Variant F showed alterations in traits that have been shown to be important for rhizosphere colonization, such as siderophore, cyanide, and exoprotease production, and these phenotypes were complemented by a cloned gacA. Sequence analysis of the gacA alelle in variant F suggested selection of the phenotype in the rhizosphere. Variant F was also affected in other phenotypes, such as lipopolysaccharide structure and flocculation in unshaken liquid medium, which were not complemented by the gacA or gacS gene. Mutation of the F113 sss gene, encoding a site-specific recombinase, showed that most of the phenotypic variation was due to the activity of this recombinase, indicating that phase variation occurs during rhizosphere colonization. PMID- 11872711 TI - raiIR genes are part of a quorum-sensing network controlled by cinI and cinR in Rhizobium leguminosarum. AB - Analysis of N-acyl-L-homoserine lactones (AHLs) produced by Rhizobium leguminosarum bv. viciae indicated that there may be a network of quorum-sensing regulatory systems producing multiple AHLs in this species. Using a strain lacking a symbiosis plasmid, which carries some of the quorum-sensing genes, we isolated mutations in two genes (raiI and raiR) that are required for production of AHLs. The raiIR genes are located adjacent to dad genes (involved in D-alanine catabolism) on a large indigenous plasmid. RaiR is predicted to be a typical LuxR type quorum-sensing regulator and is required for raiI expression. The raiR gene was expressed at a low level, possibly from a constitutive promoter, and its expression was increased under the influence of the upstream raiI promoter. Using gene fusions and analysis of AHLs produced, we showed that expression of raiI is strongly reduced in strains carrying mutations in cinI or cinR, genes which determine a higher-level quorum-sensing system that is required for normal expression of raiIR. The product of CinI, N-(3-hydroxy-7-cis tetradecenoyl) homoserine lactone, can induce raiR-dependent raiI expression, although higher levels of expression are induced by other AHLs. Expression of raiI in a strain of Agrobacterium that makes no AHLs resulted in the identification of N-(3 hydroxyoctanoyl)-L-homoserine lactone (3OH,C(8)-HSL) as the major product of RaiI, although other AHLs that comigrate with N-hexanoyl-, N-heptanoyl-, and N octanoyl-homoserine lactones were also made at low levels. The raiI gene was strongly induced by 3OH,C(8)-HSL (the product of RaiI) but could also be induced by other AHLs, suggesting that the raiI promoter can be activated by other quorum sensing systems within a network of regulation which also involves AHLs determined by genes on the symbiotic plasmid. Thus, the raiIR and cinIR genes are part of a complex regulatory network that influences AHL biosynthesis in R. leguminosarum. PMID- 11872712 TI - Osmoregulation of dimer resolution at the plasmid pJHCMW1 mwr locus by Escherichia coli XerCD recombination. AB - Xer-mediated dimer resolution at the mwr site of plasmid pJHCMW1 is osmoregulated in Escherichia coli. Whereas under low-salt conditions, the site-specific recombination reaction is efficient, under high-salt conditions, it proceeds inefficiently. Regulation of dimer resolution is independent of H-NS and is mediated by changes in osmolarity rather than ionic effects. The low level of recombination at high salt concentrations can be overcome by high levels of PepA or by mutating the ARG box to a sequence closer to the E. coli ARG box consensus. The central region of the mwr core recombination site plays a role in regulation of site-specific recombination by the osmotic pressure of the medium. PMID- 11872713 TI - VanT, a homologue of Vibrio harveyi LuxR, regulates serine, metalloprotease, pigment, and biofilm production in Vibrio anguillarum. AB - Vibrio anguillarum possesses at least two N-acylhomoserine lactone (AHL) quorum sensing circuits, one of which is related to the luxMN system of Vibrio harveyi. In this study, we have cloned an additional gene of this circuit, vanT, encoding a V. harveyi LuxR-like transcriptional regulator. A V. anguillarum Delta vanT null mutation resulted in a significant decrease in total protease activity due to loss of expression of the metalloprotease EmpA, but no changes in either AHL production or virulence. Additional genes positively regulated by VanT were identified from a plasmid-based gene library fused to a promoterless lacZ. Three lacZ fusions (serA::lacZ, hpdA-hgdA::lacZ, and sat-vps73::lacZ) were identified which exhibited decreased expression in the Delta vanT strain. SerA is similar to 3-phosphoglycerate dehydrogenases and catalyzes the first step in the serine glycine biosynthesis pathway. HgdA has identity with homogentisate dioxygenases, and HpdA is homologous to 4-hydroxyphenylpyruvate dioxygenases (HPPDs) involved in pigment production. V. anguillarum strains require an active VanT to produce high levels of an L-tyrosine-induced brown color via HPPD, suggesting that VanT controls pigment production. Vps73 and Sat are related to Vibrio cholerae proteins encoded within a DNA locus required for biofilm formation. A V. anguillarum Delta vanT mutant and a mutant carrying a polar mutation in the sat vps73 DNA locus were shown to produce defective biofilms. Hence, a new member of the V. harveyi LuxR transcriptional activator family has been characterized in V. anguillarum that positively regulates serine, metalloprotease, pigment, and biofilm production. PMID- 11872714 TI - A ToxR homolog from Vibrio anguillarum serotype O1 regulates its own production, bile resistance, and biofilm formation. AB - ToxR, a transmembrane regulatory protein, has been shown to respond to environmental stimuli. To better understand how the aquatic bacterium Vibrio anguillarum, a fish pathogen, responds to environmental signals that may be necessary for survival in the aquatic and fish environment, toxR and toxS from V. anguillarum serotype O1 were cloned. The deduced protein sequences were 59 and 67% identical to the Vibrio cholerae ToxR and ToxS proteins, respectively. Deletion mutations were made in each gene and functional analyses were done. Virulence analyses using a rainbow trout model showed that only the toxR mutant was slightly decreased in virulence, indicating that ToxR is not a major regulator of virulence factors. The toxR mutant but not the toxS mutant was 20% less motile than the wild type. Like many regulatory proteins, ToxR was shown to negatively regulate its own expression. Outer membrane protein (OMP) preparations from both mutants indicated that ToxR and ToxS positively regulate a 38-kDa OMP. The 38-kDa OMP was shown to be a major OMP, which cross-reacted with an antiserum to OmpU, an outer membrane porin from V. cholerae, and which has an amino terminus 75% identical to that of OmpU. ToxR and to a lesser extent ToxS enhanced resistance to bile. Bile in the growth medium increased expression of the 38-kDa OMP but did not affect expression of ToxR. Interestingly, a toxR mutant forms a better biofilm on a glass surface than the wild type, suggesting a new role for ToxR in the response to environmental stimuli. PMID- 11872715 TI - TonB interacts with nonreceptor proteins in the outer membrane of Escherichia coli. AB - The Escherichia coli TonB protein serves to couple the cytoplasmic membrane proton motive force to active transport of iron-siderophore complexes and vitamin B(12) across the outer membrane. Consistent with this role, TonB has been demonstrated to participate in strong interactions with both the cytoplasmic and outer membranes. The cytoplasmic membrane determinants for that interaction have been previously characterized in some detail. Here we begin to examine the nature of TonB interactions with the outer membrane. Although the presence of the siderophore enterochelin (also known as enterobactin) greatly enhanced detectable cross-linking between TonB and the outer membrane receptor, FepA, the absence of enterochelin did not prevent the localization of TonB to the outer membrane. Furthermore, the absence of FepA or indeed of all the iron-responsive outer membrane receptors did not alter this association of TonB with the outer membrane. This suggested that TonB interactions with the outer membrane were not limited to the TonB-dependent outer membrane receptors. Hydrolysis of the murein layer with lysozyme did not alter the distribution of TonB, suggesting that peptidoglycan was not responsible for the outer membrane association of TonB. Conversely, the interaction of TonB with the outer membrane was disrupted by the addition of 4 M NaCl, suggesting that these interactions were proteinaceous. Subsequently, two additional contacts of TonB with the outer membrane proteins Lpp and, putatively, OmpA were identified by in vivo cross-linking. These contacts corresponded to the 43-kDa and part of the 77-kDa TonB-specific complexes described previously. Surprisingly, mutations in these proteins individually did not appear to affect TonB phenotypes. These results suggest that there may be multiple redundant sites where TonB can interact with the outer membrane prior to transducing energy to the outer membrane receptors. PMID- 11872716 TI - RecA Protein from the extremely radioresistant bacterium Deinococcus radiodurans: expression, purification, and characterization. AB - The RecA protein of Deinococcus radiodurans (RecA(Dr)) is essential for the extreme radiation resistance of this organism. The RecA(Dr) protein has been cloned and expressed in Escherichia coli and purified from this host. In some respects, the RecA(Dr) protein and the E. coli RecA (RecA(Ec)) proteins are close functional homologues. RecA(Dr) forms filaments on single-stranded DNA (ssDNA) that are similar to those formed by the RecA(Ec). The RecA(Dr) protein hydrolyzes ATP and dATP and promotes DNA strand exchange reactions. DNA strand exchange is greatly facilitated by the E. coli SSB protein. As is the case with the E. coli RecA protein, the use of dATP as a cofactor permits more facile displacement of bound SSB protein from ssDNA. However, there are important differences as well. The RecA(Dr) protein promotes ATP- and dATP-dependent reactions with distinctly different pH profiles. Although dATP is hydrolyzed at approximately the same rate at pHs 7.5 and 8.1, dATP supports an efficient DNA strand exchange only at pH 8.1. At both pHs, ATP supports efficient DNA strand exchange through heterologous insertions but dATP does not. Thus, dATP enhances the binding of RecA(Dr) protein to ssDNA and the displacement of ssDNA binding protein, but the hydrolysis of dATP is poorly coupled to DNA strand exchange. The RecA(Dr) protein thus may offer new insights into the role of ATP hydrolysis in the DNA strand exchange reactions promoted by the bacterial RecA proteins. In addition, the RecA(Dr) protein binds much better to duplex DNA than the RecA(Ec) protein, binding preferentially to double-stranded DNA (dsDNA) even when ssDNA is present in the solutions. This may be of significance in the pathways for dsDNA break repair in Deinococcus. PMID- 11872717 TI - TrwD, the hexameric traffic ATPase encoded by plasmid R388, induces membrane destabilization and hemifusion of lipid vesicles. AB - TrwD, a hexameric ATP hydrolase encoded by plasmid R388, is a member of the PulE/VirB11 protein superfamily of traffic ATPases. It is essential for plasmid conjugation, particularly for expression of the conjugative W pilus. In the present study, we analyzed the effects that TrwD produced on unilamellar vesicles consisting of cardiolipin and phosphatidylcholine in equimolar amounts. TrwD induced dose-dependent vesicle aggregation and intervesicular mixing of the lipids located in the outer monolayers in the presence of calcium. It also induced extensive leakage of the vesicular aqueous contents. A point mutant of TrwD with a mutation in the P loop of the nucleotide-binding region (K203Q) that lacks both ATPase activity and the ability to support conjugation showed the same behavior as native TrwD in all of these processes, which were independent of the presence of ATP. Structure prediction methods revealed a close similarity to Helicobacter pylori protein HP0525, another member of the PulE/VirB11 family, whose crystal structure is known. The interpretation of our data in the light of this structure is that TrwD interacts with the lipid bilayer through hydrophobic regions in its N-terminal domain, which leads to a certain degree of membrane destabilization. TrwD appears to be a part of the conjugation machinery that interacts with the membranous systems in order to facilitate DNA transfer in bacteria. PMID- 11872718 TI - Extensive variation in the O-antigen gene cluster within one Salmonella enterica serogroup reveals an unexpected complex history. AB - The 46 serogroups of Salmonella enterica have different O-antigens, and each is thought to have a specific form of the O-antigen cluster. Comparison of the 145 serovars of serogroup B revealed much more intraserogroup genetic diversity than expected. The O27 factor, due to an alpha 1-6 linkage between O units in place of the more common alpha 1-2 linkage and previously thought to be due to a converting bacteriophage, is now shown to be due to a wzy(alpha(1-6)) gene located within the major gene cluster. Surprisingly a remnant of this gene in all O27(-) serovars shows that the ancestor was O27(+). There are six distinct gene cluster forms, five apparently derived by a series of deletions and one by an insertion from an ancestral O27(+) form present in 57 serovars. The history of the gene cluster and movement between subspecies I and II can be traced. Two of the derivative forms still have a functional wzy(alpha(1-6)) gene, while in three it has been inactivated by deletion or insertion. Two of the forms lacking a functional wzy(alpha(1-6)) gene have the wzy(alpha(1-2)) gene first described for strain LT2 as rfc, whereas for the third the wzy gene has not been located. PMID- 11872719 TI - An extracellular matrix-associated zinc metalloprotease is required for dilauroyl phosphatidylethanolamine chemotactic excitation in Myxococcus xanthus. AB - An extracellular matrix connects bacteria that live in organized assemblages called biofilms. While the role of the matrix in the regulation of cell behavior has not been extensively examined in bacteria, we suggest that, like mammalian cells, the matrix facilitates cell-cell interactions involved with regulation of cohesion, motility, and sensory transduction. The extracellular matrix of the soil bacterium Myxococcus xanthus is essential for biofilm formation and fruiting body development. The matrix material is extruded as long, thin fibrils that mediate adhesion to surfaces, cohesion to other cells, and excitation by the chemoattractant dilauroyl phosphatidylethanolamine. We report the identification of a putative matrix-associated zinc metalloprotease called FibA (fibril protein A). Western blotting with FibA-specific monoclonal antibody 2105 suggests extensive proteolytic processing of FibA during assembly into fibrils, consistent with the autoprocessing observed with other members of the M4 metalloprotease family. Disruption of fibA had no obvious effect on the structure of the fibrils and did not inhibit cell cohesion, excitation by dioleoyl phosphatidylethanolamine, or activity of the A- or S-motility motors. However, the cells lost the ability to respond to dilauroyl phosphatidylethanolamine and to form well-spaced fruiting bodies, though substantial aggregation was observed. Chemotactic excitation of the fibA mutant was restored by incubation with purified wild-type fibrils. The results suggest that this metalloprotease is involved in sensory transduction. PMID- 11872720 TI - Oxygen-mediated regulation of porphobilinogen formation in Rhodobacter capsulatus. AB - A Rhodobacter capsulatus hemC mutant has been isolated and used to show that oxygen regulates the intracellular levels of porphobilinogen. Experiments using a hemB-cat gene fusion demonstrated that oxygen does not transcriptionally regulate hemB transcription. Porphobilinogen synthase activity is not regulated by oxygen nor is the enzyme feedback inhibited by hemin or protoporphyrin IX. It was demonstrated that less than 20% of [(14)C]aminolevulinate was incorporated into bacteriochlorophyll, suggesting that the majority of the aminolevulinate is diverted from the common tetrapyrrole pathway. Porphobilinogen oxygenase activity was not observed in this organism; however, an NADPH-linked aminolevulinate dehydrogenase activity was demonstrated. The specific activity of this enzyme increased with increasing oxygen tension. The results presented here suggest that carbon flow over the common tetrapyrrole pathway is regulated by a combination of feedback inhibition of aminolevulinate synthase and diversion of aminolevulinate from the pathway by aminolevulinate dehydrogenase. PMID- 11872721 TI - Regulation of nap gene expression and periplasmic nitrate reductase activity in the phototrophic bacterium Rhodobacter sphaeroides DSM158. AB - Bacterial periplasmic nitrate reductases (Nap) can play different physiological roles and are expressed under different conditions depending on the organism. Rhodobacter sphaeroides DSM158 has a Nap system, encoded by the napKEFDABC gene cluster, but nitrite formed is not further reduced because this strain lacks nitrite reductase. Nap activity increases in the presence of nitrate and oxygen but is unaffected by ammonium. Reverse transcription-PCR and Northern blots demonstrated that the napKEFDABC genes constitute an operon transcribed as a single 5.5-kb product. Northern blots and nap-lacZ fusions revealed that nap expression is threefold higher under aerobic conditions but is regulated by neither nitrate nor ammonium, although it is weakly induced by nitrite. On the other hand, nitrate but not nitrite causes a rapid enzyme activation, explaining the higher Nap activity found in nitrate-grown cells. Translational nap'-'lacZ fusions reveal that the napK and napD genes are not efficiently translated, probably due to mRNA secondary structures occluding the translation initiation sites of these genes. Neither butyrate nor caproate increases nap expression, although cells growing phototrophically on these reduced substrates show a very high Nap activity in vivo (nitrite accumulation is sevenfold higher than in medium with malate). Phototrophic growth on butyrate or caproate medium is severely reduced in the NapA(-) mutants. Taken together, these results indicate that nitrate reduction in R. sphaeroides is mainly regulated at the level of enzyme activity by both nitrate and electron supply and confirm that the Nap system is involved in redox balancing using nitrate as an ancillary oxidant to dissipate excess reductant. PMID- 11872722 TI - Two different lantibiotic-like peptides originate from the ericin gene cluster of Bacillus subtilis A1/3. AB - A lantibiotic gene cluster was identified in Bacillus subtilis A1/3 showing a high degree of homology to the subtilin gene cluster and occupying the same genetic locus as the spa genes in B. subtilis ATCC 6633. The gene cluster exhibits diversity with respect to duplication of two subtilin-like genes which are separated by a sequence similar to a portion of a lanC gene. Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) analyses of B. subtilis A1/3 culture extracts confirmed the presence of two lantibiotic-like peptides, ericin S (3,442 Da) and ericin A (2,986 Da). Disruption of the lanB-homologous gene eriB resulted in loss of production of both peptides, demonstrating that they are processed in an eriB-dependent manner. Although precursors of ericins S and A show only 75% of identity, the matured lantibiotic-like peptides reveal highly similar physical properties; separation was only achieved after multistep, reversed-phase high-performance liquid chromatography. Based on Edman and peptidase degradation in combination with MALDI-TOF MS, for ericin S a subtilin-like, lanthionine-bridging pattern is supposed. For ericin A two C-terminal rings are different from the lanthionine pattern of subtilin. Due to only four amino acid exchanges, ericin S and subtilin revealed similar antibiotic activities as well as similar properties in response to heat and protease treatment. For ericin A only minor antibiotic activity was found. PMID- 11872723 TI - IntI2 integron integrase in Tn7. AB - Integrons can insert and excise antibiotic resistance genes on plasmids in bacteria by site-specific recombination. Class 1 integrons code for an integrase, IntI1 (337 amino acids in length), and are generally borne on elements derived from Tn5090, such as that found in the central part of Tn21. A second class of integron is found on transposon Tn7 and its relatives. We have completed the sequence of the Tn7 integrase gene, intI2, which contains an internal stop codon. This codon was found to be conserved among intI2 genes on three other Tn7-like transposons harboring different cassettes. The predicted peptide sequence (IntI2*) is 325 amino acids long and is 46% identical to IntI1. In order to detect recombination activity, the internal stop codon at position 179 in the parental allele was changed to a triplet coding for glutamic acid. The sequences flanking the cassette arrays in the class 1 and 2 integrons are not closely related, but a common pool of mobile cassettes is used by the different integron classes; two of the three antibiotic resistance cassettes on Tn7 and its close relatives are also found in various class 1 integrons. We also observed a fourth excisable cassette downstream of those described previously in Tn7. The fourth cassette encodes a 165-amino-acid protein of unknown function with 6.5 contiguous repeats of a sequence coding for 7 amino acids. IntI2*179E promoted site-specific excision of each of the cassettes in Tn7 at different frequencies. The integrases from Tn21 and Tn7 showed limited cross-specificity in that IntI1 could excise all cassettes from both Tn21 and Tn7. However, we did not observe a corresponding excision of the aadA1 cassette from Tn21 by IntI2*179E. PMID- 11872724 TI - Rubredoxins involved in alkane oxidation. AB - Rubredoxins (Rds) are essential electron transfer components of bacterial membrane-bound alkane hydroxylase systems. Several Rd genes associated with alkane hydroxylase or Rd reductase genes were cloned from gram-positive and gram negative organisms able to grow on n-alkanes (Alk-Rds). Complementation tests in an Escherichia coli recombinant containing all Pseudomonas putida GPo1 genes necessary for growth on alkanes except Rd 2 (AlkG) and sequence comparisons showed that the Alk-Rds can be divided in AlkG1- and AlkG2-type Rds. All alkane degrading strains contain AlkG2-type Rds, which are able to replace the GPo1 Rd 2 in n-octane hydroxylation. Most strains also contain AlkG1-type Rds, which do not complement the deletion mutant but are highly conserved among gram-positive and gram-negative bacteria. Common to most Rds are the two iron-binding CXXCG motifs. All Alk-Rds possess four negatively charged residues that are not conserved in other Rds. The AlkG1-type Rds can be distinguished from the AlkG2-type Rds by the insertion of an arginine downstream of the second CXXCG motif. In addition, the glycines in the two CXXCG motifs are usually replaced by other amino acids. Mutagenesis of residues conserved in either the AlkG1- or the AlkG2-type Rds, but not between both types, shows that AlkG1 is unable to transfer electrons to the alkane hydroxylase mainly due to the insertion of the arginine, whereas the exchange of the glycines in the two CXXCG motifs only has a limited effect. PMID- 11872725 TI - Functional analysis of alkane hydroxylases from gram-negative and gram-positive bacteria. AB - We have cloned homologs of the Pseudomonas putida GPo1 alkane hydroxylase from Pseudomonas aeruginosa PAO1, Pseudomonas fluorescens CHA0, Alcanivorax borkumensis AP1, Mycobacterium tuberculosis H37Rv, and Prauserella rugosa NRRL B 2295. Sequence comparisons show that the level of protein sequence identity between the homologs is as low as 35%, and that the Pseudomonas alkane hydroxylases are as distantly related to each other as to the remaining alkane hydroxylases. Based on the observation that rubredoxin, an electron transfer component of the GPo1 alkane hydroxylase system, can be replaced by rubredoxins from other alkane hydroxylase systems, we have developed three recombinant host strains for the functional analysis of the novel alkane hydroxylase genes. Two hosts, Escherichia coli GEc137 and P. putida GPo12, were equipped with pGEc47 Delta B, which encodes all proteins necessary for growth on medium-chain-length alkanes (C(6) to C(12)), except a functional alkane hydroxylase. The third host was an alkB knockout derivative of P. fluorescens CHA0, which is no longer able to grow on C(12) to C(16) alkanes. All alkane hydroxylase homologs, except the Acinetobacter sp. ADP1 AlkM, allowed at least one of the three hosts to grow on n alkanes. PMID- 11872726 TI - Partitioning of chromosomal DNA during establishment of cellular asymmetry in Bacillus subtilis. AB - The switch from symmetric to asymmetric cell division is a key feature of development in many organisms, including Bacillus subtilis sporulation. Here we demonstrate that, prior to the onset of asymmetric cell division, the B. subtilis chromosome is partitioned into two unequally sized domains, with the origin proximal one-third of the future forespore chromosome condensed near one pole of the cell. Asymmetric chromosome partitioning is independent of polar division, as it occurs in cells depleted of FtsZ but depends on two transcription factors that govern the initiation of sporulation, sigma(H) and Spo0A-P. It is also independent of chromosome partitioning proteins Spo0J and Soj, suggesting the existence of a novel mechanism controlling chromosome structure. Thus, our results demonstrate that, during sporulation, two separable events prepare B. subtilis for asymmetric cell division: the relocation of cell division sites to the cell poles and the asymmetric partitioning of the future forespore chromosome. PMID- 11872727 TI - Glyoxylate regeneration pathway in the methylotroph Methylobacterium extorquens AM1. AB - Most serine cycle methylotrophic bacteria lack isocitrate lyase and convert acetyl coenzyme A (acetyl-CoA) to glyoxylate via a novel pathway thought to involve butyryl-CoA and propionyl-CoA as intermediates. In this study we have used a genome analysis approach followed by mutation to test a number of genes for involvement in this novel pathway. We show that methylmalonyl-CoA mutase, an R-specific crotonase, isobutyryl-CoA dehydrogenase, and a GTPase are involved in glyoxylate regeneration. We also monitored the fate of (14)C-labeled carbon originating from acetate, butyrate, or bicarbonate in mutants defective in glyoxylate regeneration and identified new potential intermediates in the pathway: ethylmalonyl-CoA, methylsuccinyl-CoA, isobutyryl-CoA, methacrylyl-CoA, and beta-hydroxyisobutyryl-CoA. A new scheme for the pathway is proposed based on these data. PMID- 11872728 TI - A two-component regulator mediates population-density-dependent expression of the Bradyrhizobium japonicum nodulation genes. AB - Bradyrhizobium japonicum nod gene expression was previously shown to be population density dependent. Induction of the nod genes is highest at low culture density and repressed at high population densities. This repression involves both NolA and NodD2 and is mediated by an extracellular factor found in B. japonicum conditioned medium. NolA and NodD2 expression is maximal at high population densities. We demonstrate here that a response regulator, encoded by nwsB, is required for the full expression of the B. japonicum nodYABC operon. In addition, NwsB is also required for the population-density-dependent expression of both nolA and nodD2. Expression of nolA and nodD2 in the nwsB mutant remained at a basal level, even at high culture densities. The nwsB defect could be complemented by overexpression of a second response regulator, NodW. Consistent with the fact that NolA and NodD2 repress nod gene expression, the expression of a nodY-lacZ fusion in the nwsB mutant was unaffected by culture density. In plant assays with GUS fusions, nodules infected with the wild type showed no nodY-GUS expression. In contrast, nodY-GUS expression was not repressed in nodules infected with the nwsB mutant. Nodule competition assays between the wild type and the nwsB mutant revealed that the addition of conditioned medium resulted in a competitive advantage for the nwsB mutant. PMID- 11872729 TI - Analyses of the roles of the three cheA homologs in chemotaxis of Vibrio cholerae. AB - The Vibrio cholerae genome revealed the presence of multiple sets of chemotaxis genes, including three cheA gene homologs. We found that the cheA-2, but not cheA 1 or cheA-3, gene is essential for chemotaxis under standard conditions. Loss of chemotaxis had no effect on virulence factor expression in vitro. PMID- 11872730 TI - Heat shock proteome of Agrobacterium tumefaciens: evidence for new control systems. AB - The regulation of Agrobacterium tumefaciens heat shock genes involves a transcriptional activator (RpoH) and repressor elements (HrcA-CIRCE). Using proteome analysis and mutants in these control elements, we show that the heat shock induction of 32 (out of 56) heat shock proteins is independent of RpoH and HrcA. These results indicate the existence of additional regulatory factors in the A. tumefaciens heat shock response. PMID- 11872731 TI - Identification of XcpZ domains required for assembly of the secreton of Pseudomonas aeruginosa. AB - Most of the exoproteins secreted by Pseudomonas aeruginosa are transported via the type II secretion system. This machinery, which is widely conserved in gram negative bacteria, consists of 12 Xcp proteins organized as a multiprotein complex, also called the secreton. We previously reported that the mutual stabilization of XcpZ and XcpY plays an important role in the assembly of the secreton. In this study, we engineered variant XcpZ proteins by using linker insertion mutagenesis. We identified three distinct regions of XcpZ required for both the stabilization of XcpY and the functionality of the secreton. Interestingly, we also demonstrated that another component of the machinery, XcpP, can modulate the stabilizing activity of XcpZ on XcpY. PMID- 11872732 TI - Induction of ResDE-dependent gene expression in Bacillus subtilis in response to nitric oxide and nitrosative stress. AB - Transcription of ResDE-controlled genes in Bacillus subtilis was induced by sodium nitroprusside and nitric oxide. This induction requires the sensor kinase ResE and the response regulator ResD. Among members of the ResDE regulon, only the flavohemoglobin gene was induced by nitrosative stress via both a ResDE dependent mechanism and an unidentified ResDE-independent mechanism. PMID- 11872734 TI - Lack of regulation of the modification-dependent restriction enzyme McrBC in Escherichia coli. AB - Restriction alleviation (RA) by the type I restriction enzyme EcoKI is caused by treatments that damage DNA. RA is due to proteolysis of the EcoKI HsdR subunit by the ClpXP ATP-dependent protease. Here we show that the modification-dependent enzyme McrBC is not subject to RA, although it is moderately sensitive to ClpAP. PMID- 11872733 TI - The active partition gene incC of IncP plasmids is required for stable maintenance in a broad range of hosts. AB - Plasmids of incompatibility group P (IncP) are capable of replication and stable inheritance in a wide variety of gram-negative bacteria. Three determinants of IncP plasmids are components of an active partition locus that is predicted to function in the segregation of plasmid copies to daughter cells. These determinants are incC, which codes for a member of the ParA family of partition ATPases; korB, which specifies a DNA-binding protein that also functions as a global transcriptional repressor; and O(B), the DNA target for KorB, which occurs at multiple locations on IncP plasmids. To determine the importance and host range of the IncC/KorB partition system in the maintenance of IncP plasmids, we constructed an in-frame deletion of incC in the otherwise intact 60-kb IncP alpha plasmid R995. R995 Delta incC was found to be highly unstable in Escherichia coli, Pseudomonas aeruginosa, Pseudomonas putida, Agrobacterium tumefaciens, and Acinetobacter calcoaceticus, whereas wild-type R995 is stable in all these hosts. In addition, R995 Delta incC could not be established in Actinobacillus actinomycetemcomitans. trans-Complementation analysis showed that the coding region for IncC2 polypeptide, which is expressed from an internal translational start within the incC gene, was sufficient to restore stable maintenance to wild type levels. The results show that the IncC/KorB active partition system of IncP plasmids is remarkably proficient for stable maintenance in diverse bacteria. PMID- 11872735 TI - Site-directed mutagenesis studies of selected motif and charged residues and of cysteines of the multifunctional tetracycline efflux protein Tet(L). AB - All of the transmembrane glutamates of Tet(L) are essential for tetracycline (TET) resistance, and E397 has been shown to be essential for all catalytic modes, i.e., TET-Me(2+) and Na(+) efflux and K(+) uptake. Loop residues D74 and G70 are essential for TET flux but not for Na(+) or K(+) flux. A cysteineless Tet(L) protein exhibits all activities. PMID- 11872736 TI - The tra region of the conjugative plasmid pIP501 is organized in an operon with the first gene encoding the relaxase. AB - The tra genes orf1 to orf11 of pIP501 were shown to be transcribed as a single operon of 11.3 kb in Enterococcus faecalis by reverse transcription-PCR. The transcriptional start site of the tra mRNA was mapped at 110 bp upstream from the predicted TTG start codon of the first gene of the operon, the traA relaxase. The TraA protein (660 amino acids) and a C-terminally truncated version of the TraA protein (293 amino acids) were purified as fusions with glutathione S transferase. oriT cleavage activity of both TraA proteins was demonstrated in vitro on supercoiled plasmid pVA2241 DNA containing oriT(pIP501). The activity of the DNA relaxase TraA is strictly dependent on the presence of Mg(2+) or Mn(2+) and is highest at temperatures of between 42 and 45C. PMID- 11872737 TI - Protective role of tolC in efflux of the electron shuttle anthraquinone-2,6 disulfonate. AB - Extracellular electron transfer can play an important role in microbial respiration on insoluble minerals. The humic acid analog anthraquinone-2,6 disulfonate (AQDS) is commonly used as an electron shuttle during studies of extracellular electron transfer. Here we provide genetic evidence that AQDS enters Shewanella oneidensis strain MR-1 and causes cell death if it accumulates past a critical concentration. A tolC homolog protects the cell from toxicity by mediating the efflux of AQDS. Electron transfer to AQDS appears to be independent of the tolC pathway, however, and requires the outer membrane protein encoded by mtrB. We suggest that there may be structural and functional relationships between quinone-containing electron shuttles and antibiotics. PMID- 11872738 TI - The IntI-like tyrosine recombinase of Shewanella oneidensis is active as an integron integrase. AB - We have found an integron-like integrase gene and an attI site in Shewanella oneidensis as part of a small chromosomal integron. We have cloned this gene and tested the ability of the integrase to excise cassettes from various integrons. Most cassettes flanked by two attC sites are readily excised, while cassettes in the "first" position, with an attI1 or attI3 site on one end, are not excised. An exception is a cassette with attI2 on one end. The attI2 site, from Tn7, has greater similarity to the attI site adjacent to the integrase of S. oneidensis than do attI1 or attI3. We cloned the attI site of S. oneidensis and observed the integration of two different cassettes. We have, therefore, demonstrated the function of this integron-like integrase. PMID- 11872739 TI - Interdomain interaction of Stat3 regulates its Src homology 2 domain-mediated receptor binding activity. AB - Activation of Stat proteins by cytokines is initiated by their Src homology 2 (SH2) domain-mediated association with the cytokine receptors. Previously, we identified an essential role of the coiled-coil domain of Stat3 in binding of the receptor peptides derived from the interleukin-6 receptor subunit, gp130. In this study, we further investigated the molecular basis of this regulation. We found that the C-terminal domain of Stat3 negatively regulates its receptor binding activity only in the absence of the first alpha-helix of the coiled-coil domain, which leads to a hypothesis of intramolecular interaction. Physical interactions between the coiled-coil domain and the C-terminal domain, as well as the SH2 domain, were indeed detected. Furthermore, a sub-region of the C-terminal domain (amino acids 720-740), which is also involved in the interaction with the coiled coil domain, was demonstrated to be critical for the regulation of the receptor binding. Correspondingly, phosphorylation on Ser-727 within this region inhibits this interaction. In agreement with the peptide binding results, both the coiled coil domain and the C-terminal sub-region are necessary for the functional recruitment of Stat3 to the cellular gp130 in response to interleukin-6, suggesting that the interdomain interaction is a prerequisite for the SH2 mediated receptor binding in interleukin-6 signaling. PMID- 11872740 TI - Coaggregation, cointernalization, and codesensitization of adenosine A2A receptors and dopamine D2 receptors. AB - Antagonistic and reciprocal interactions are known to exist between adenosine and dopamine receptors in the striatum. In the present study, double immunofluorescence experiments with confocal laser microscopy showed a high degree of colocalization of adenosine A(2A) receptors (A(2A)R) and dopamine D(2) receptors (D(2)R) in cell membranes of SH-SY5Y human neuroblastoma cells stably transfected with human D(2)R and in cultured striatal cells. A(2A)R/D(2)R heteromeric complexes were demonstrated in coimmunoprecipitation experiments in membrane preparations from D(2)R-transfected SH-SY5Y cells and from mouse fibroblast Ltk(-) cells stably transfected with human D(2)R (long form) and transiently cotransfected with the A(2A)R double-tagged with hemagglutinin. Long term exposure to A(2A)R and D(2)R agonists in D(2)R-cotransfected SH-SY5Y cells resulted in coaggregation, cointernalization and codesensitization of A(2A)R and D(2)R. These results give a molecular basis for adenosine-dopamine antagonism at the membrane level and have implications for treatment of Parkinson's disease and schizophrenia, in which D(2)R are involved. PMID- 11872742 TI - Enzymatic properties of S-adenosylmethionine synthetase from the archaeon Methanococcus jannaschii. AB - S-Adenosylmethionine synthetase (ATP:l-methionine S-adenosyltransferase, MAT) catalyzes a unique enzymatic reaction that leads to formation of the primary biological alkylating agent. MAT from the hyperthermophilic archaeon Methanococcus jannaschii (MjMAT) is a prototype of the newly discovered archaeal class of MAT proteins that are nearly unrecognizable in sequence when compared with the class that encompasses both the eucaryal and bacterial enzymes. In this study the functional properties of purified recombinant MjMAT have been evaluated. The products of the reaction are AdoMet, PP(i), and P(i); >90% of the P(i) originates from the gamma-phosphoryl group of ATP. The circular dichroism spectrum of the dimeric MjMAT indicates that the secondary structure is more helical than the Escherichia coli counterpart (EcMAT), suggesting a different protein topology. The steady state kinetic mechanism is sequential, with random addition of ATP and methionine; AdoMet is the first product released, followed by release of PP(i) and P(i). The substrate specificity differs remarkably from the previously characterized MATs; the nucleotide binding site has a very broad tolerance of alterations in the adenosine moiety. MjMAT has activity at 70 degrees C comparable with that of EcMAT at 37 degrees C, consistent with the higher temperature habitat of M. jannaschii. The activation energy for AdoMet formation is larger than that for the E. coli MAT-catalyzed reaction, in accord with the notion that enzymes from thermophilic organisms are often more rigid than their mesophilic counterparts. The broad substrate tolerance of this enzyme proffers routes to preparation of novel AdoMet analogs. PMID- 11872741 TI - Interactions of phocein with nucleoside-diphosphate kinase, Eps15, and Dynamin I. AB - Phocein, an intracellular protein interacting with striatin, bears a few homologies with the sigma-subunits of clathrin adaptor proteins (Baillat, G., Moqrich, A., Castets, F., Baude, A., Bailly, Y., Benmerah, A., and Monneron, A. (2001) Mol. Biol. Cell 12, 663-673). Using phocein as a bait in a yeast two hybrid screen, we identified two novel interacting proteins, nucleoside diphosphate kinase (NDPK) and Eps15. Immunoprecipitation and pull-down experiments involving native and/or recombinant phocein and, respectively, NDPK and Eps15, biochemically validated their interactions. NDPK and Eps15 were recently shown to be functional neighbors of dynamin. Dynamin I is shown here to directly interact with NDPK through its C-terminal proline-rich domain, whereas recombinant phocein associates with native dynamin I. Immunocytochemical studies of rat embryonic hippocampal neurons demonstrated partial co-localization of phocein and dynamin I. Phocein thus appears to be a component of the complexes involved in some steps of the vesicular traffic machinery. PMID- 11872743 TI - A proton pump ATPase with testis-specific E1-subunit isoform required for acrosome acidification. AB - The vacuolar-type H(+)-ATPases (V-ATPases) are a family of multimeric proton pumps involved in a wide variety of physiological processes. We have identified two novel mouse genes, Atp6e1 and Atp6e2, encoding testis-specific (E1) and ubiquitous (E2) V-ATPase subunit E isoforms, respectively. The E1 transcript appears about 3 weeks after birth, corresponding to the start of meiosis, and is expressed specifically in round spermatids in seminiferous tubules. Immunohistochemistry with isoform-specific antibodies revealed that the V-ATPase with E1 and a2 isoforms is located specifically in developing acrosomes of spermatids and acrosomes in mature sperm. In contrast, the E2 isoform was expressed in all tissues examined and present in the perinuclear compartments of spermatocytes. The E1 isoform exhibits 70% identity with the E2, and both isoforms functionally complemented a null mutation of the yeast counterpart VMA4, indicating that they are bona fide V-ATPase subunits. The chimeric enzymes showed slightly lower K(m)(ATP) than yeast V-ATPase. Consistent with the temperature sensitive growth of Deltavma4-expressing E1 isoform, vacuolar membrane vesicles exhibited temperature-sensitive coupling between ATP hydrolysis and proton transport. These results suggest that E1 isoform is essential for energy coupling involved in acidification of acrosome. PMID- 11872744 TI - Partially folded structure of flavin adenine dinucleotide-depleted ferredoxin NADP+ reductase with residual NADP+ binding domain. AB - Maize ferredoxin-NADP(+) reductase (FNR) consists of flavin adenine dinucleotide (FAD) and NADP(+) binding domains with a FAD molecule bound noncovalently in the cleft between these domains. The structural changes of FNR induced by dissociation of FAD have been characterized by a combination of optical and biochemical methods. The CD spectrum of the FAD-depleted FNR (apo-FNR) suggested that removal of FAD from holo-FNR produced an intermediate conformational state with partially disrupted secondary and tertiary structures. Small angle x-ray scattering indicated that apo-FNR assumes a conformation that is less globular in comparison with holo-FNR but is not completely chain-like. Interestingly, the replacement of tyrosine 95 responsible for FAD binding with alanine resulted in a molecular form similar to apo-protein of the wild-type enzyme. Both apo- and Y95A FNR species bound to Cibacron Blue affinity resin, indicating the presence of a native-like conformation for the NADP(+) binding domain. On the other hand, no evidence was found for the existence of folded conformations in the FAD binding domains of these proteins. These results suggested that FAD-depleted FNR assumes a partially folded structure with a residual NADP(+) binding domain but a disordered FAD binding domain. PMID- 11872745 TI - Involvement of VIP36 in intracellular transport and secretion of glycoproteins in polarized Madin-Darby canine kidney (MDCK) cells. AB - VIP36, an intracellular lectin that recognizes high mannose-type glycans (Hara Kuge, S., Ohkura, T., Seko, A., and Yamashita, K. (1999) Glycobiology 9, 833 839), was shown to localize not only to the early secretory pathway but also to the plasma membrane of Madin-Darby canine kidney (MDCK) cells. In the plasma membrane, VIP36 exhibited an apical-predominant distribution, the apical/basolateral ratio being approximately 2. Like VIP36, plasma membrane glycoproteins recognized by VIP36 were found in the apical and basolateral membranes in the ratio of approximately 2 to 1. In addition, secretory glycoproteins recognized by VIP36 were secreted approximately 2-fold more efficiently from the apical membrane than from the basolateral membrane. Thus, the apical/basolateral ratio of the transport of VIP36-recognized glycoproteins was correlated with that of VIP36 in MDCK cells. Upon overproduction of VIP36 in MDCK cells, the apical/basolateral ratios of both VIP36 and VIP36-recognized glycoproteins were changed from approximately 2 to approximately 4, and the secretion of VIP36-recognized glycoproteins was greatly stimulated. In contrast to the overproduction of VIP36, that of a mutant version of VIP36, which has no lectin activity, was of no effect on the distribution of glycoproteins to apical and basolateral membranes and inhibited the secretion of VIP36-recognized glycoproteins. Furthermore, the overproduction of VIP36 greatly stimulated the secretion of a major apical secretory glycoprotein of MDCK cells, clusterin, which was found to carry at least one high mannose-type glycan and to be recognized by VIP36. In contrast to the secretion of clusterin, that of a non glycosylated apical-secretion protein, galectin-3, was not stimulated through the overproduction of VIP36. These results indicated that VIP36 was involved in the transport and sorting of glycoproteins carrying high mannose-type glycan(s). PMID- 11872746 TI - Tyrosine kinase c-Src constitutes a bridge between cystic fibrosis transmembrane regulator channel failure and MUC1 overexpression in cystic fibrosis. AB - Cystic fibrosis (CF), a disease caused by mutations in the cystic fibrosis transmembrane regulator (CFTR) chloride channel, is associated in the respiratory system with the accumulation of mucus and impaired lung function. The role of the CFTR channel in the regulation of the intracellular pathways that determine the overexpression of mucin genes is unknown. Using differential display, we have observed the differential expression of several mRNAs that may correspond to putative CFTR-dependent genes. One of these mRNAs was further characterized, and it corresponds to the tyrosine kinase c-Src. Additional results suggest that c Src is a central element in the pathway connecting the CFTR channel with MUC1 overexpression and that the overexpression of mucins is a primary response to CFTR malfunction in cystic fibrosis, which occurs even in the absence of bacterial infection. PMID- 11872747 TI - Signal pathways involved in activation of p70S6K and phosphorylation of 4E-BP1 following exposure of multiple myeloma tumor cells to interleukin-6. AB - Interleukin-6 (IL-6) is a prominent tumor growth factor for malignant multiple myeloma cells. In addition to its known activation of the Janus tyrosine kinase STAT and RAS-MEK-ERK pathways, recent work suggests that IL-6 can also activate the phosphatidylinositol 3-kinase (PI3-K)/AKT kinase pathway in myeloma cells. Because activation of the PI3-K/AKT as well as RAS-MEK-ERK pathways may result in downstream stimulation of the p70(S6K) (p70) and phosphorylation of the 4E-BP1 translational repressor, we assessed these potential molecular targets in IL-6 treated myeloma cells. IL-6 rapidly activated p70 kinase activity and p70 phosphorylation. Activation was inhibited by wortmannin, rapamycin, and the ERK inhibitors PD98059 and UO126, as well as by a dominant negative mutant of AKT. The concurrent requirements for both ERK and PI3-K/AKT appeared to be a result of their ability to phosphorylate p70 on different residues. In contrast, IL-6 induced phosphorylation of 4E-BP1 was inhibited by rapamycin, wortmannin, and dominant negative AKT but ERK inhibitors had no effect, indicating ERK function was dispensable. In keeping with these data, a dominant active AKT mutant was sufficient to induce 4E-BP1 phosphorylation but could not by itself activate p70 kinase activity. Prevention of IL-6-induced p70 activation and 4E-BP1 phosphorylation by the mammalian target of rapamycin inhibitors rapamycin and CCI 779 resulted in inhibition of IL-6-induced myeloma cell growth. These results indicate that both ERK and PI3-K/AKT pathways are required for optimal IL-6 induced p70 activity, but PI3-K/AKT is sufficient for 4E-BP1 phosphorylation. Both effects are mediated via mammalian target of rapamycin function, and, furthermore, these effects are critical for IL-6-induced tumor cell growth. PMID- 11872748 TI - NF-kappa B as a therapeutic target in multiple myeloma. AB - We have shown that thalidomide (Thal) and its immunomodulatory derivatives (IMiDs), proteasome inhibitor PS-341, and As(2)O(3) act directly on multiple myeloma (MM) cells and in the bone marrow (BM) milieu to overcome drug resistance. Although Thal/IMiDs, PS-341, and As(2)O(3) inhibit nuclear factor (NF)-kappaB activation, they also have multiple and varied other actions. In this study, we therefore specifically address the role of NF-kappaB blockade in mediating anti-MM activity. To characterize the effect of specific NF-kappaB blockade on MM cell growth and survival in vitro, we used an IkappaB kinase (IKK) inhibitor (PS-1145). Our studies demonstrate that PS-1145 and PS-341 block TNFalpha-induced NF-kappaB activation in a dose- and time-dependent fashion in MM cells through inhibition of IkappaBalpha phosphorylation and degradation of IkappaBalpha, respectively. Dexamethasone (Dex), which up-regulates IkappaBalpha protein, enhances blockade of NF-kappaB activation by PS-1145. Moreover, PS-1145 blocks the protective effect of IL-6 against Dex-induced apotosis. TNFalpha induced intracellular adhesion molecule (ICAM)-1 expression on both RPMI8226 and MM.1S cells is also inhibited by PS-1145. Moreover, PS-1145 inhibits both IL-6 secretion from BMSCs triggered by MM cell adhesion and proliferation of MM cells adherent to BMSCs. However, in contrast to PS-341, PS-1145 only partially (20 50%) inhibits MM cell proliferation, suggesting that NF-kappaB blockade cannot account for all of the anti-MM activity of PS-341. Importantly, however, TNFalpha induces MM cell toxicity in the presence of PS-1145. These studies demonstrate that specific targeting of NF-kappaB can overcome the growth and survival advantage conferred both by tumor cell binding to BMSCs and cytokine secretion in the BM milieu. Furthermore, they provide the framework for clinical evaluation of novel MM therapies based upon targeting NF-kappaB. PMID- 11872749 TI - Calbindin D28k exhibits properties characteristic of a Ca2+ sensor. AB - Calbindin D(28k) is a member of the calmodulin superfamily of Ca(2+)-binding proteins and contains six EF-hands. The protein is generally believed to function as a Ca(2+) buffer, but the studies presented in this work indicate that it may also act as a Ca(2+) sensor. The results show that Mg(2+) binds to the same sites as Ca(2+) with an association constant of approximately 1.4.10(3) m(-1) in 0.15 m KCl. The four high affinity sites in calbindin D(28k) bind Ca(2+) in a non sequential, parallel manner. In the presence of physiological concentrations of Mg(2+), the Ca(2+) affinity is reduced by a factor of 2, and the cooperativity, which otherwise is modest, increases. Based on the binding constants determined in the presence of physiological salt concentrations, we estimate that at the Ca(2+) concentration in a resting cell calbindin D(28k) is saturated to 40-75% with Mg(2+) but to less than 9% with Ca(2+). In contrast, the protein is expected to be nearly fully saturated with Ca(2+) at the Ca(2+) level of an activated cell. A substantial conformational change is observed upon Ca(2+) binding, but only minor structural changes take place upon Mg(2+) binding. This suggests that calbindin D(28k) undergoes Ca(2+)-induced structural changes upon Ca(2+) activation of a cell. Thus, calbindin D(28k) displays several properties that would be expected for a protein involved in Ca(2+)-induced signal transmission and hence may function not only as a Ca(2+) buffer but also as a Ca(2+) sensor. Digestion patterns resulting from limited proteolysis of the protein suggest that the loop of EF-hand 2, a variant site that does not bind Ca(2+), becomes exposed upon Ca(2+) binding. PMID- 11872750 TI - Ubiquitin (UbC) expression in muscle cells is increased by glucocorticoids through a mechanism involving Sp1 and MEK1. AB - The muscle protein catabolism present in rats with insulin-dependent diabetes and other catabolic conditions is generally associated with increased glucocorticoid production and mRNAs encoding components of the ubiquitin-proteasome system. The mechanisms that increase ubiquitin (UbC) expression have not been identified. We studied the regulation of UbC expression in L6 muscle cells because dexamethasone stimulates the transcription of this gene and others encoding components of the ubiquitin-proteasome pathway. Results of in vivo genomic DNA footprinting experiments indicate that a protein(s) binds to Sp1 sites approximately 50 bp upstream from the UbC transcription start site; dexamethasone changes the methylation pattern at these sites. Sp1 binds to DNA probes corresponding to the rat or human UbC promoter, and treating cells with dexamethasone increases this binding. Deletion and mutation analyses of the rat and human UbC promoters are consistent with an important role of Sp1 in UbC induction by glucocorticoids. Dexamethasone-induced ubiquitin expression is blocked by mithramycin, an inhibitor of Sp1 binding. UO126, a pharmacologic inhibitor of MEK1, also blocks UbC transcriptional activation by dexamethasone; L6 cells transfected to express constitutively active MEK1 exhibit increased UbC promoter activity. Thus, glucocorticoids increase UbC expression in muscle cells by a novel transcriptional mechanism involving Sp1 and MEK1. PMID- 11872751 TI - A secreted type of beta 1,6-N-acetylglucosaminyltransferase V (GnT-V) induces tumor angiogenesis without mediation of glycosylation: a novel function of GnT-V distinct from the original glycosyltransferase activity. AB - Angiogenesis is the first regulatory step of tumor progression. Herein, we report on some findings that show that beta1,6-N-acetylglucosaminyltransferase V (GnT-V) functions as an inducer of angiogenesis that has a novel and completely different function from the original function of glycosyltransferase. A secreted type of GnT-V protein itself promoted angiogenesis in vitro and in vivo at physiological concentrations. The highly basic domain of GnT-V induced the release of fibroblast growth factor-2 from heparan sulfate proteoglycan on the cell surface and/or extracellular matrix, leading to angiogenesis. These findings provide some novel information on the relationship between GnT-V and tumor metastasis. The inhibition of GnT-V secretion or its expression represents a novel potential strategy for the inhibition of tumor angiogenesis. PMID- 11872752 TI - Active-site residues governing high steroid isomerase activity in human glutathione transferase A3-3. AB - Glutathione transferase (GST) A3-3 is the most efficient human steroid double bond isomerase known. The activity with Delta(5)-androstene-3,17-dione is highly dependent on the phenolic hydroxyl group of Tyr-9 and the thiolate of glutathione. Removal of these groups caused an 1.1 x 10(5)-fold decrease in k(cat); the Y9F mutant displayed a 150-fold lower isomerase activity in the presence of glutathione and a further 740-fold lower activity in the absence of glutathione. The Y9F mutation in GST A3-3 did not markedly decrease the activity with the alternative substrate 1-chloro-2,4-dinitrobenzene. Residues Phe-10, Leu 111, and Ala-216 selectively govern the activity with the steroid substrate. Mutating residue 111 into phenylalanine caused a 25-fold decrease in k(cat)/K(m) for the steroid isomerization. The mutations A216S and F10S, separate or combined, affected the isomerase activity only marginally, but with the additional L111F mutation k(cat)/K(m) was reduced to 0.8% of that of the wild type value. In contrast, the activities with 1-chloro-2,4-dinitrobenzene and phenethylisothiocyanate were not largely affected by the combined mutations F10S/L111F/A216S. K(i) values for Delta(5)-androstene-3,17-dione and Delta(4) androstene-3,17-dione were increased by the triple mutation F10S/L111F/A216S. The pK(a) of the thiol group of active-site-bound glutathione, 6.1, increased to 6.5 in GST A3-3/Y9F. The pK(a) of the active-site Tyr-9 was 7.9 for the wild-type enzyme. The pH dependence of k(cat)/K(m) of wild-type GST A3-3 for the isomerase reaction displays two kinetic pK(a) values, 6.2 and 8.1. The basic limb of the pH dependence of k(cat) and k(cat)/K(m) disappears in the Y9F mutant. Therefore, the higher kinetic pK(a) reflects ionization of Tyr-9, and the lower one reflects ionization of glutathione. We propose a reaction mechanism for the double-bond isomerization involving abstraction of a proton from C4 in the steroid accompanied by protonation of C6, the thiolate of glutathione serving as a base and Tyr-9 assisting by polarizing the 3-oxo group of the substrate. PMID- 11872753 TI - The multivalent PDZ domain-containing protein CIPP is a partner of acid-sensing ion channel 3 in sensory neurons. AB - Acid-sensing ion channels (ASICs) are cationic channels activated by extracellular pH. They are present in the brain, where they are thought to participate in signal transduction associated with local pH variations, and in sensory neurons, where they have been involved in pain perception associated with tissue acidosis and in mechanoperception. The ASIC3 subunit is mainly expressed in dorsal root ganglion neurons. Its expression is associated with a rapidly inactivating current followed by a slowly activating sustained current thought to be required for the tonic sensation of pain caused by acids. We report here the interaction of this channel subunit with the multivalent PDZ (PSD-95 Drosophila discs-large protein, Zonula occludens protein 1) domain-containing protein CIPP. This interaction requires the C-terminal region of ASIC3 and the fourth PDZ domain of CIPP. Co-expression of CIPP and ASIC3 in COS cells increases the maximal ASIC3 peak current density by a factor of 5 and slightly shifts the pH(0.5) for activation from pH 6.2 to pH 6.4. CIPP mRNA is found at a significant level in the same dorsal root ganglion neuronal cell population that expresses the ASIC3 subunit, i.e. mainly in the small nociceptive neurons. CIPP is thus a scaffolding protein that could both enhance the surface expression of ASIC3 and bring together ASIC3 and functionally related proteins in the membrane of sensory neurons. PMID- 11872754 TI - Growth-dependent regulation of mammalian pyrimidine biosynthesis by the protein kinase A and MAPK signaling cascades. AB - The carbamoyl phosphate synthetase domain of the multifunctional protein CAD catalyzes the initial, rate-limiting step in mammalian de novo pyrimidine biosynthesis. In addition to allosteric regulation by the inhibitor UTP and the activator PRPP, the carbamoyl phosphate synthetase activity is controlled by mitogen-activated protein kinase (MAPK)- and protein kinase A (PKA)-mediated phosphorylation. MAPK phosphorylation, both in vivo and in vitro, increases sensitivity to PRPP and decreases sensitivity to the inhibitor UTP, whereas PKA phosphorylation reduces the response to both allosteric effectors. To elucidate the factors responsible for growth state-dependent regulation of pyrimidine biosynthesis, the activity of the de novo pyrimidine pathway, the MAPK and PKA activities, the phosphorylation state, and the allosteric regulation of CAD were measured as a function of growth state. As cells entered the exponential growth phase, there was an 8-fold increase in pyrimidine biosynthesis that was accompanied by a 40-fold increase in MAPK activity and a 4-fold increase in CAD threonine phosphorylation. PRPP activation increased to 21-fold, and UTP became a modest activator. These changes were reversed when the cultures approach confluence and growth ceases. Moreover, CAD phosphoserine, a measure of PKA phosphorylation, increased 2-fold in confluent cells. These results are consistent with the activation of CAD by MAPK during periods of rapid growth and its down-regulation in confluent cells associated with decreased MAPK phosphorylation and a concomitant increase in PKA phosphorylation. A scheme is proposed that could account for growth-dependent regulation of pyrimidine biosynthesis based on the sequential action of MAPK and PKA on the carbamoyl phosphate synthetase activity of CAD. PMID- 11872756 TI - Osvald T. Avery and the Nobel Prize in medicine. PMID- 11872755 TI - Thiol-disulfide oxidoreductases are essential for the production of the lantibiotic sublancin 168. AB - Thiol-disulfide oxidoreductases are required for disulfide bond formation in proteins that are exported from the cytoplasm. Four enzymes of this type, termed BdbA, BdbB, BdbC, and BdbD, have been identified in the Gram-positive eubacterium Bacillus subtilis. BdbC and BdbD have been shown to be critical for the folding of a protein required for DNA uptake during natural competence. In contrast, no function has been assigned so far to the BdbA and BdbB proteins. The bdbA and bdbB genes are located in one operon that also contains the genes specifying the lantibiotic sublancin 168 and the ATP-binding cassette transporter SunT. Interestingly sublancin 168 contains two disulfide bonds. The present studies demonstrate that SunT and BdbB, but not BdbA, are required for the production of active sublancin 168. In addition, the BdbB paralogue BdbC is at least partly able to replace BdbB in sublancin 168 production. These observations show the unprecedented involvement of thiol-disulfide oxidoreductases in the synthesis of a peptide antibiotic. Notably BdbB cannot complement BdbC in competence development, showing that these two closely related thiol-disulfide oxidoreductases have different, but partly overlapping, substrate specificities. PMID- 11872757 TI - Tetanus prophylaxis in the A&E department. PMID- 11872758 TI - The decline of rehabilitation services and its impact on disability benefits. PMID- 11872759 TI - A clutch of new syndromes? PMID- 11872760 TI - Emergency ambulance dispatch: is there a case for triage? AB - Emergency telephone calls for an ambulance (999 calls) are usually dealt with first-come first-served. We have devised and assessed criteria that ambulance dispatch might use to prioritize responses. Data were collected retrospectively on consecutive patients presenting to an accident and emergency (A&E) department after a 999 call. An unblinded researcher abstracted data including age, date, time, caller, location, reason for call and A&E diagnosis and each case was examined for ten predetermined criteria necessitating an immediate ambulance response--namely, cardiac arrest; chest pain; shortness of breath; altered mental status/seizure; abdominal/loin pain >65 years old; fresh haematemesis; fall >2m; stabbing; major burns. 471 patients were recruited, 55% male, median age 50 years. 406 calls came from bystanders or the patients themselves, 36 from general practitioners, 8 from other hospitals and 21 from the police. 52% of patients were admitted. 44% met at least one of the above criteria. Most patients did not meet the criteria for an immediate ambulance response but might nonetheless be suitable for an urgent response. The criteria used in this study have the advantage of being based on the history provided by the caller. The introduction of a priority-based dispatch system could reduce response times to those who are seriously ill, and also improve road safety. PMID- 11872761 TI - Venous thromboembolic prophylaxis for transurethral prostatectomy: practice among British urologists. AB - Venous thromboembolism (VTE) is an occasional cause of death after transurethral prostatectomy but there are no established guidelines for its prevention in relation to this operation. We assessed practice in the UK by mailing a questionnaire to 460 consultant members of the British Association of Urological Surgeons. 362 (79%) completed questionnaires were received. 280 of 362 (77%) respondents routinely used VTE prophylaxis with transurethral prostatectomy; 82 (23%) did not. 230 of the 280 urologists who took precautions used mechanical methods; 50 used low dose heparin, either with stockings or alone. This survey indicates that, despite a lack of clear evidence, most British urologists favour some form of precaution against VTE in patients undergoing transurethral prostatectomy. PMID- 11872762 TI - Wheeze and Mycoplasma pneumoniae. PMID- 11872763 TI - Refractory coeliac disease, small-bowel lymphoma and chorea. PMID- 11872764 TI - Bilateral knee pain with hyperparathyroidism. PMID- 11872765 TI - Perineal necrotizing fasciitis with dilatation of Cowper's gland. PMID- 11872766 TI - The man who walks backwards. PMID- 11872767 TI - Cholecystitis after cholecystectomy. PMID- 11872769 TI - Understanding organizational culture in reforming the National Health Service. PMID- 11872770 TI - Geophagia: the history of earth-eating. PMID- 11872771 TI - Medicine and the Spanish Civil War. PMID- 11872774 TI - Secondary tumours of the penis. PMID- 11872772 TI - Sir John Franklin's last arctic expedition: a medical disaster. PMID- 11872773 TI - Personal experiences in the malaria eradication campaign 1955-1962. PMID- 11872775 TI - Crash. PMID- 11872776 TI - Revalidation of the retired. PMID- 11872777 TI - Revalidation of the retired. PMID- 11872778 TI - Revalidation of the retired. PMID- 11872779 TI - Do in-hospital waiting lists show self-regulation? PMID- 11872780 TI - Edge of chaos. PMID- 11872781 TI - Control of foot and mouth disease. PMID- 11872782 TI - Counselling and consent in vasectomy. PMID- 11872783 TI - Edge of chaos. PMID- 11872785 TI - The art of the mentally ill. PMID- 11872787 TI - The health of frequent business travellers. PMID- 11872788 TI - The prevalence of musculoskeletal troubles among car drivers. AB - In order to explore the relationship between car driving and musculoskeletal troubles, a cross-sectional structured-interview survey of low- to high-mileage drivers (including individuals who drove as part of their job) was conducted based on the Nordic Musculoskeletal Questionnaire. The results clearly showed that exposure to car driving was associated with reported sickness absence due to low back trouble and that those who drive as part of their job appear to be more at risk from low back trouble than those whose jobs primarily involve sitting (not driving) and standing activities. The frequency of reported discomfort also increased with higher annual mileage. In addition, drivers of cars with more adjustable driving packages had fewer reported musculoskeletal troubles. This identifies an urgent need for the training of managers of fleet vehicles in the importance of developing measures to reduce this problem, for example, the selection of an individual's car with respect to comfort and postural criteria. PMID- 11872789 TI - Fragrance as an occupational allergen. AB - Sensitization to fragrance is believed to occur mainly outside the workplace. This study addresses the frequency of fragrance allergy in patch test patients of differing occupations during a 15 year period. The occupation most strongly associated with fragrance allergy in both sexes was health care work (positive tests in 11.7% of males and 10.4% of females). Retired individuals also had high rates of fragrance allergy (11.6% of males and 14.5% of females), and the prevalence of sensitization increased with advancing age. Health care workers and metal workers had statistically significantly higher rates of allergy to eugenol than did workers in other occupations. Food handlers had significantly higher rates of allergy to cinnamal and cinnamic alcohol. These findings suggest that sensitization to fragrance occurs more frequently in an occupational setting than is generally understood and could have implications for preventive measures. PMID- 11872790 TI - Guidance on standards of health for clinical health care workers. AB - Judgements about the health of clinical health care workers in relation to fitness to practice are made by a variety of doctors. These guidelines have been written to assist with such judgements and to facilitate equitable decision making in matters of employment. PMID- 11872792 TI - Prevalence of hepatitis B and C virus infection in barbers in the Sivas region of Turkey. AB - Hepatitis B virus (HBV) and hepatitis C virus (HCV) infections are among the most devastating health problems in the world, including Turkey. The route of transmission of HBV and HCV is mainly parenteral, a small number of epidemiological studies demonstrating that perinatal, sexual, household and occupational transmission occurs. Contact of a patient's blood or bodily fluids with non-intact skin is another mode of HBV and HCV transmission. Barbers in Turkey may often be exposed accidentally to the blood and bodily fluids of their customers. The aim of this study was to determine the prevalence of HBV and HCV infection in barbers. We conducted a study to determine the prevalence of antibodies against HBV and HCV among 176 barbers and 180 control subjects in the Sivas region of Turkey. The prevalence of HBV and HCV was found to be higher in barbers (39.8 and 2.8%, respectively) than in a comparison group (28.3 and 1.1%, respectively). No significant relationship was found with the duration of occupation. Among the seropositive subjects, it was found that most had been exposed to needle pricks or scissor cuts. Our data suggest that both HBV and HCV infections may constitute occupational hazards for barbers. The sources of infection could be not only such personal risk factors as 'sharps' injuries and scissor cuts, but may also include other unknown factors. PMID- 11872791 TI - Four worksite weight loss programs with high-stress occupations using a meal replacement product. AB - The objective was to evaluate the efficacy of worksite weight reduction programs at high-stress worksites. We employed a longitudinal study based on two meal replacements daily with subjects choosing a third 'sensible' meal. The subjects were 492 healthy, overweight men and women working in high-stress occupations (police, hospital health professionals, flight crew members, firefighters). The mean group ages ranged from 32.17 +/- 5.70 to 44.50 +/- 16.40 years; the mean group body mass indexes (BMIs) ranged from 27.40 +/- 2.54 to 32.90 +/- 3.39 kg/m(2). The completion rate for the 12 weeks was 79.8%. Reductions in mean weight and mean BMI were greater than in medically supervised clinical trials with non-worksite adults. Firefighters lost the most weight and medical personnel the least. Follow-up found considerable retention of weight loss. Men lost significantly more weight than women (p < 0.006). We conclude that employees in some high-stress settings may participate productively in worksite weight reduction and maintenance programs that use meal replacements. PMID- 11872793 TI - Epidemiological surveillance at Electricite de France-Gaz de France: health assessment of nuclear power plant employees between 1993 and 1998. AB - Because the 17,500 employees working in nuclear power plants at Electricite de France, the national power company, may be exposed to a wide variety of industrial hazards, the health insurance department of the company has set up an epidemiological surveillance programme for them. This report describes its first stage, an analysis of their health problems. This descriptive, cross-sectional and exhaustive study examined sick-leave, mortality and cancer incidence to assess the health of the employees working from 1993 through 1998. The analysis compared the employees in nuclear power plants, considered 'exposed', with the rest of the personnel of Electricite de France-Gaz de France, the 'non-exposed' (125,000 persons). Relative risks for these variables were estimated, after stratification for age, sex and work grade. Globally, the employees in the nuclear sector appeared to have fewer health problems than the other company employees. This was true regardless of age and especially for men, operating employees and supervisory employees. Nonetheless, three points must be noted: non work accidents generated a non-significant excess of absenteeism and mortality among these employees, especially among management and supervisory personnel; suicides affected supervisors in particular; and an excess of primary malignant brain tumours affected both mortality [relative risk (RR) = 1.96, n.s.] and incidence, especially among operating employees (RR = 2.87, 95% confidence interval = 1.00-8.43). No excess of malignant blood disease was observed. This study, performed for surveillance purposes, points to brain tumours as a problem that raises important aetiological questions. Research is now necessary to provide answers. Surveillance of nuclear plant employees at Electricite de France should be continued and expanded to include the study of specific occupational exposures. PMID- 11872795 TI - Lead exposure in scaffolders during refurbishment construction activity--an observational study. AB - The toxic effects of lead have been known for centuries. Occupational exposure to this chemical hazard has also been well documented in relation to various industry groups, including construction, where workers are recognized as being significantly exposed during refurbishment work, in particular through inhalation and ingestion of lead fumes and dust. It is easy to see how so-called 'burners', 'cutters' and 'blasters'--workers directly involved in removing old lead paint- may become exposed; the influence of personal hygiene, smoking, eating/drinking and nail biting has also been documented in the literature. We now report on one group, the scaffolders, not previously considered to be at risk. Although not directly involved in the paint removal, anecdotal and personal experience of the authors indicate that these workers, who erect and later dismantle access structures during the renovation of previously lead-painted surfaces, may take up significant amounts of lead, mainly by ingestion, to raise their personal blood lead levels (and body burden) in line with recognized 'lead workers'. Exposures of this magnitude would also bring the scaffolders involved in such refurbishment work under the Control of Lead at Work Regulations 1998. The authors make various recommendations on measures to minimize and control exposure of scaffolders to lead. PMID- 11872794 TI - Maxillofacial injuries related to work accidents: a new concept of a hospital based full electronic occupational trauma surveillance system. AB - The objective of this study was to investigate the occurrence of occupational maxillofacial injuries using a newly installed relational database. Twenty-six injuries were identified out of 28,624 patients admitted to the emergency unit during a 12 month period. Falls from height or being struck by moving objects were common causes of these injuries. Two-thirds of those identified were construction workers. This paper demonstrates the power of modern databases to identify specific occurrences that may provide the basis for prevention in the future. PMID- 11872797 TI - Papers that have changed the practice of occupational medicine: 'Pulmonary disease due to inhalation of derivatives of Bacillus subtilis containing proteolytic enzyme' by M.L.H. Flindt. 1969. PMID- 11872798 TI - Chemically-induced hepatitis in a Taiwanese male construction worker. PMID- 11872801 TI - Noble gas-actinide compounds: complexation of the CUO molecule by Ar, Kr, and Xe atoms in noble gas matrices. AB - The CUO molecule, formed from the reaction of laser-ablated U atoms with CO in a noble gas, exhibits very different stretching frequencies in a solid argon matrix [804.3 and 852.5 wave numbers (cm(-1))] than in a solid neon matrix (872.2 and 1047.3 cm(-1)). Related experiments in a matrix consisting of 1% argon in neon suggest that the argon atoms are interacting directly with the CUO molecule. Relativistic density functional calculations predict that CUO can bind directly to one argon atom (U-Ar = 3.16 angstroms; binding energy = 3.2 kilocalories per mole), accompanied by a change in the ground state from a singlet to a triplet. Our experimental and theoretical results also suggest that multiple argon atoms can bind to a single CUO molecule. PMID- 11872800 TI - Living with lethal PIP3 levels: viability of flies lacking PTEN restored by a PH domain mutation in Akt/PKB. AB - The phosphoinositide phosphatase PTEN is mutated in many human cancers. Although the role of PTEN has been studied extensively, the relative contributions of its numerous potential downstream effectors to deregulated growth and tumorigenesis remain uncertain. We provide genetic evidence in Drosophila melanogaster for the paramount importance of the protein kinase Akt [also called protein kinase B (PKB)] in mediating the effects of increased phosphatidylinositol 3,4,5 trisphosphate (PIP3) concentrations that are caused by the loss of PTEN function. A mutation in the pleckstrin homology (PH) domain of Akt that reduces its affinity for PIP3 sufficed to rescue the lethality of flies devoid of PTEN activity. Thus, Akt appears to be the only critical target activated by increased PIP3 concentrations in Drosophila. PMID- 11872802 TI - Requirement for a peptidoglycan recognition protein (PGRP) in Relish activation and antibacterial immune responses in Drosophila. AB - Components of microbial cell walls are potent activators of innate immune responses in animals. For example, the mammalian TLR4 signaling pathway is activated by bacterial lipopolysaccharide and is required for resistance to infection by Gram-negative bacteria. Other components of microbial surfaces, such as peptidoglycan, are also potent activators of innate immune responses, but less is known about how those components activate host defense. Here we show that a peptidoglycan recognition protein, PGRP-LC, is absolutely required for the induction of antibacterial peptide genes in response to infection in Drosophila and acts by controlling activation of the NF-kappaB family transcription factor Relish. PMID- 11872803 TI - Beauty and the beast. PMID- 11872804 TI - Disappointing data scuttle plans for large-scale AIDS vaccine trial. PMID- 11872805 TI - Has GM corn 'invaded' Mexico? PMID- 11872806 TI - NAS asks for more scrutiny of GM crops. PMID- 11872807 TI - T. rex was no runner, muscle study shows. PMID- 11872808 TI - Solar system kicks up its own dust. PMID- 11872809 TI - Reforms would loosen bonds, cut safety net. PMID- 11872810 TI - NASA decision not suited for women. PMID- 11872811 TI - CDC head's resignation expands leadership void. PMID- 11872812 TI - Battle heats up over mammography benefits. PMID- 11872813 TI - Clear-cut publication rules prove elusive. PMID- 11872814 TI - Forest biotech edges out of the lab. PMID- 11872815 TI - Can genetic engineering help restore 'heritage' trees? PMID- 11872816 TI - New state of matter not so new? PMID- 11872817 TI - Elbow room and new jewelry. PMID- 11872818 TI - Viral threat to newborns under radar. PMID- 11872819 TI - 'Earth Simulator' puts Japan on the cutting edge. PMID- 11872820 TI - Tangled roots? Genetics meets genealogy. PMID- 11872821 TI - Portrait of Science. Scientist, technologist, proto-feminist, superstar. AB - Although Marie Curie is known primarily for her discovery of radium, her true gift to science was her realization that radioactivity is an intrinsic atomic property of matter rather than the result of chemical processes. She was one of the few Nobel laureates to win the prize twice (physics and chemistry). During her career and as one of the first prominent women scientists, she became increasingly aware of the need for funding for research and of the scientific freedom that money can bring. By nature shy and reserved, Marie's fame, as both a scientist and as an exemplar of a liberated professional woman of the roaring twenties, grew to superstar proportions. PMID- 11872822 TI - Does BDNF have pre- or postsynaptic targets? AB - A type of synaptic plasticity in the brain called long-term potentiation (LTP) is thought to form the molecular basis of learning and memory. In a Perspective, Manabe discusses new findings (Kovalchuk et al.) showing brain-derived neurotropic factor modulates LTP by binding to TrkB receptors on the postsynaptic neuron. PMID- 11872823 TI - Blood out of a stone. AB - Almost 50 years ago, astronomers provided evidence for powerful radio emissions localized in two giant "lobes" on opposite sides of elliptical galaxies. These lobes are now known to be powered by jets emanating from black holes at the centers of the galaxies. In his Perspective, Blandford discusses how the black hole may generate the jets. He highlights the report by Koide et al., who have simulated one mechanism for supplying the jets with energy via a magnetic connection to the gas just outside the black hole's event horizon. PMID- 11872824 TI - A pit stop at the ER. AB - Although ligand activation of receptor signaling is well understood, less is known about how a cell switches off signaling by the activated receptor. In his Perspective, Gill discusses new work (Haj et al.) that visualizes one step in the process of deactivating a ligand-activated receptor tyrosine kinase--the dephosphorylation of the internalized receptor by a phosphatase in the endoplasmic reticulum. PMID- 11872825 TI - Making a network of hydrophobic clusters. AB - The early events in protein folding are often difficult to track. A hydrophobic collapse, during which nonpolar residues are buried away from the polar solvent, has been proposed, but little is known about the role of this collapse in protein folding. In his Perspective, Baldwin discusses the report by Klein-Seetharaman et al., who shed some light on this issue. A network of hydrophobic clusters stabilizes at least one nonnative interaction in unfolded hen lysozyme. In conjunction with earlier studies, the results suggest that this network has a beneficial effect on the folding of the protein. PMID- 11872826 TI - Close before opening. AB - As bacteria need iron from the environment to survive, they have evolved active iron transporter proteins in their outer membranes. In her Perspective, Postle discusses new insights into iron transport revealed by the crystal structure of the iron transporter FecA in E. coli (Ferguson et al.). PMID- 11872827 TI - The next frontier. AB - High-intensity lasers are opening up a new realm of light-matter interactions. In his Perspective, Yamanouchi reviews recent progress in this field, focusing on two intensity regimes: the Coulombic regime, which mostly deforms molecular structures and causes tunneling ionization, and the relativistic regime, where high-intensity lasers produce x-rays and high-energy particles and may cause nuclear fusion reactions. Efforts are under way to increase laser intensity further for accessing the next frontier. PMID- 11872829 TI - Systems biology: a brief overview. AB - To understand biology at the system level, we must examine the structure and dynamics of cellular and organismal function, rather than the characteristics of isolated parts of a cell or organism. Properties of systems, such as robustness, emerge as central issues, and understanding these properties may have an impact on the future of medicine. However, many breakthroughs in experimental devices, advanced software, and analytical methods are required before the achievements of systems biology can live up to their much-touted potential. PMID- 11872830 TI - Reverse engineering of biological complexity. AB - Advanced technologies and biology have extremely different physical implementations, but they are far more alike in systems-level organization than is widely appreciated. Convergent evolution in both domains produces modular architectures that are composed of elaborate hierarchies of protocols and layers of feedback regulation, are driven by demand for robustness to uncertain environments, and use often imprecise components. This complexity may be largely hidden in idealized laboratory settings and in normal operation, becoming conspicuous only when contributing to rare cascading failures. These puzzling and paradoxical features are neither accidental nor artificial, but derive from a deep and necessary interplay between complexity and robustness, modularity, feedback, and fragility. This review describes insights from engineering theory and practice that can shed some light on biological complexity. PMID- 11872832 TI - Modeling the heart--from genes to cells to the whole organ. AB - Successful physiological analysis requires an understanding of the functional interactions between the key components of cells, organs, and systems, as well as how these interactions change in disease states. This information resides neither in the genome nor even in the individual proteins that genes code for. It lies at the level of protein interactions within the context of subcellular, cellular, tissue, organ, and system structures. There is therefore no alternative to copying nature and computing these interactions to determine the logic of healthy and diseased states. The rapid growth in biological databases; models of cells, tissues, and organs; and the development of powerful computing hardware and algorithms have made it possible to explore functionality in a quantitative manner all the way from the level of genes to the physiological function of whole organs and regulatory systems. This review illustrates this development in the case of the heart. Systems physiology of the 21st century is set to become highly quantitative and, therefore, one of the most computer-intensive disciplines. PMID- 11872833 TI - Flight of the dodo. PMID- 11872831 TI - A genomic regulatory network for development. AB - Development of the body plan is controlled by large networks of regulatory genes. A gene regulatory network that controls the specification of endoderm and mesoderm in the sea urchin embryo is summarized here. The network was derived from large-scale perturbation analyses, in combination with computational methodologies, genomic data, cis-regulatory analysis, and molecular embryology. The network contains over 40 genes at present, and each node can be directly verified at the DNA sequence level by cis-regulatory analysis. Its architecture reveals specific and general aspects of development, such as how given cells generate their ordained fates in the embryo and why the process moves inexorably forward in developmental time. PMID- 11872834 TI - Quantum impurities in the two-dimensional spin one-half Heisenberg antiferromagnet. AB - The study of randomness in low-dimensional quantum antiferromagnets is at the forefront of research in the field of strongly correlated electron systems, yet there have been relatively few experimental model systems. Complementary neutron scattering and numerical experiments demonstrate that the spin-diluted Heisenberg antiferromagnet La2Cu1-z(Zn,Mg)(z)O4 is an excellent model material for square lattice site percolation in the extreme quantum limit of spin one-half. Measurements of the ordered moment and spin correlations provide important quantitative information for tests of theories for this complex quantum-impurity problem. PMID- 11872835 TI - Micro/nano encapsulation via electrified coaxial liquid jets. AB - We report a method to generate steady coaxial jets of immiscible liquids with diameters in the range of micrometer/nanometer size. This compound jet is generated by the action of electro-hydrodynamic (EHD) forces with a diameter that ranges from tens of nanometers to tens of micrometers. The eventual jet breakup results in an aerosol of monodisperse compound droplets with the outer liquid surrounding or encapsulating the inner one. Following this approach, we have produced monodisperse capsules with diameters varying between 10 and 0.15 micrometers, depending on the running parameters. PMID- 11872836 TI - A thermally re-mendable cross-linked polymeric material. AB - We have developed a transparent organic polymeric material that can repeatedly mend or "re-mend" itself under mild conditions. The material is a tough solid at room temperature and below with mechanical properties equaling those of commercial epoxy resins. At temperatures above 120 degrees C, approximately 30% (as determined by solid-state nuclear magnetic resonance spectroscopy) of "intermonomer" linkages disconnect but then reconnect upon cooling, This process is fully reversible and can be used to restore a fractured part of the polymer multiple times, and it does not require additional ingredients such as a catalyst, additional monomer, or special surface treatment of the fractured interface. PMID- 11872837 TI - Niobium-zirconium chronometry and early solar system development. AB - Niobium-92 (92Nb) decays to zirconium-92 (92Zr) with a half-life of 36 million years and can be used to place constraints on the site of p-process nucleosynthesis and the timing of early solar system processes. Recent results have suggested that the initial 92Nb/93Nb of the solar system was high (>10(-3)). We report Nb-Zr internal isochrons for the ordinary chondrite Estacado (H6) and a clast of the mesosiderite Vaca Muerta, both of which define an initial 92Nb/93Nb ratio of approximately 10(-5). Therefore, the solar system appears to have started with a ratio of <3 x 10(-5), which implies that Earth's initial differentiation need not have been as protracted as recently suggested. PMID- 11872838 TI - Imaging sites of receptor dephosphorylation by PTP1B on the surface of the endoplasmic reticulum. AB - When bound by extracellular ligands, receptor tyrosine kinases (RTKs) on the cell surface transmit critical signals to the cell interior. Although signal termination is less well understood, protein tyrosine phosphatase-1B (PTP1B) is implicated in the dephosphorylation and inactivation of several RTKs. However, PTP1B resides on the cytoplasmic surface of the endoplasmic reticulum (ER), so how and when it accesses RTKs has been unclear. Using fluorescence resonance energy transfer (FRET) methods, we monitored interactions between the epidermal- and platelet-derived growth factor receptors and PTP1B. PTP1B-catalyzed dephosphorylation required endocytosis of the receptors and occurred at specific sites on the surface of the ER. Most of the RTKs activated at the cell surface showed interaction with PTP1B after internalization, establishing that RTK activation and inactivation are spatially and temporally partitioned within cells. PMID- 11872839 TI - Discrete microdomains with high concentration of cAMP in stimulated rat neonatal cardiac myocytes. AB - The second messenger cyclic adenosine monophosphate (cAMP) is the most important modulator of sympathetic control over cardiac contractility. In cardiac myocytes and many other cell types, however, cAMP transduces the signal generated upon stimulation of various receptors and activates different cellular functions, raising the issue of how specificity can be achieved. In the general field of signal transduction, the view is emerging that specificity is guaranteed by tight localization of signaling events. Here, we show that in neonatal rat cardiac myocytes, beta-adrenergic stimulation generates multiple microdomains with increased concentration of cAMP in correspondence with the region of the transverse tubule/junctional sarcoplasmic reticulum membrane. The restricted pools of cAMP show a range of action as small as approximately 1 micrometer, and free diffusion of the second messenger is limited by the activity of phosphodiesterases. Furthermore, we demonstrate that such gradients of cAMP specifically activate a subset of protein kinase A molecules anchored in proximity to the T tubule. PMID- 11872841 TI - Long-range interactions within a nonnative protein. AB - Protein folding and unfolding are coupled to a range of biological phenomena, from the regulation of cellular activity to the onset of neurodegenerative diseases. Defining the nature of the conformations sampled in nonnative proteins is crucial for understanding the origins of such phenomena. We have used a combination of nuclear magnetic resonance (NMR) spectroscopy and site-directed mutagenesis to study unfolded states of the protein lysozyme. Extensive clusters of hydrophobic structure exist within the wild-type protein even under strongly denaturing conditions. These clusters involve distinct regions of the sequence but are all disrupted by a single point mutation that replaced residue Trp62 with Gly located at the interface of the two major structural domains in the native state. Thus, nativelike structure in the denatured protein is stabilized by the involvement of Trp62 in nonnative and long-range interactions. PMID- 11872840 TI - Structural basis of gating by the outer membrane transporter FecA. AB - Siderophore-mediated acquisition systems facilitate iron uptake. We present the crystallographic structure of the integral outer membrane receptor FecA from Escherichia coli with and without ferric citrate at 2.5 and 2.0 angstrom resolution. FecA is composed of three distinct domains: the barrel, plug, and NH2 terminal extension. Binding of ferric citrate triggers a conformational change of the extracellular loops that close the external pocket of FecA. Ligand-induced allosteric transitions are propagated through the outer membrane by the plug domain, signaling the occupancy of the receptor in the periplasm. These data establish the structural basis of gating for receptors dependent on the cytoplasmic membrane protein TonB. By compiling available data for this family of receptors, we propose a mechanism for the energy-dependent transport of siderophores. PMID- 11872842 TI - A functional screen for the type III (Hrp) secretome of the plant pathogen Pseudomonas syringae. AB - Type III secreted "effector" proteins of bacterial pathogens play central roles in virulence, yet are notoriously difficult to identify. We used an in vivo genetic screen to identify 13 effectors secreted by the type III apparatus (called Hrp, for "hypersensitive response and pathogenicity") of the plant pathogen Pseudomonas syringae. Although sharing little overall homology, the amino-terminal regions of these effectors had strikingly similar amino acid compositions. This feature facilitated the bioinformatic prediction of 38 P. syringae effectors, including 15 previously unknown proteins. The secretion of two of these putative effectors was shown to be type III--dependent. Effectors showed high interstrain variation, supporting a role for some effectors in adaptation to different hosts. PMID- 11872843 TI - Colorectal cancer in mice genetically deficient in the mucin Muc2. AB - The gastrointestinal tract is lined by a layer of mucus comprised of highly glycosylated proteins called mucins. To evaluate the importance of mucin in intestinal carcinogenesis, we constructed mice genetically deficient in Muc2, the most abundant secreted gastrointestinal mucin. Muc2-/- mice displayed aberrant intestinal crypt morphology and altered cell maturation and migration. Most notably, the mice frequently developed adenomas in the small intestine that progressed to invasive adenocarcinoma, as well as rectal tumors. Thus, Muc2 is involved in the suppression of colorectal cancer. PMID- 11872844 TI - Postsynaptic Induction of BDNF-Mediated Long-Term Potentiation. AB - Brain-derived neurotrophic factor (BDNF) and other neurotrophins are critically involved in long-term potentiation (LTP). Previous reports point to a presynaptic site of neurotrophin action. By imaging dentate granule cells in mouse hippocampal slices, we identified BDNF-evoked Ca2+ transients in dendrites and spines, but not at presynaptic sites. Pairing a weak burst of synaptic stimulation with a brief dendritic BDNF application caused an immediate and robust induction of LTP. LTP induction required activation of postsynaptic Ca2+ channels and N-methyl-d-aspartate receptors and was prevented by the blockage of postsynaptic Ca2+ transients. Thus, our results suggest that BDNF-mediated LTP is induced postsynaptically. Our finding that dendritic spines are the exclusive synaptic sites for rapid BDNF-evoked Ca2+ signaling supports this conclusion. PMID- 11872846 TI - Understanding racial-ethnic and societal differentials in STI. PMID- 11872845 TI - Divided by cytochrome oxidase: a map of the projections from V1 to V2 in macaques. AB - Current models partition the primate visual system into dorsal (magno) and ventral (parvo, konio) streams. Perhaps the strongest evidence for this idea has come from the pattern of projections between the primary visual area (V1) and the second visual area (V2). Prior studies describe three distinct pathways: magno to thick stripes, parvo to pale stripes, and konio to thin stripes. We now demonstrate that V1 output arises from just two sources: patch columns and interpatch columns. Patch columns project to thin stripes and interpatch columns project to pale and thick stripes. Projection of interpatches to common V2 stripe types (pale and thick) merges parvo and magno inputs, making it likely that these functional channels are distributed strongly to both dorsal and ventral streams. PMID- 11872848 TI - Preventive human papillomavirus vaccination. PMID- 11872850 TI - An introduction to mathematical models in sexually transmitted disease epidemiology. AB - Mathematical models serve a number of roles in understanding sexually transmitted infection epidemiology and control. This article seeks to provide the non mathematician with a description of their construction and use and presents illustrative examples from sexually transmitted infection epidemiology. PMID- 11872851 TI - Progressive intrathoracic lymphadenopathy: EBV associated non-Hodgkin's lymphoma. AB - A 30 year old man presented with late stage HIV disease and intrathoracic lymphadenopathy. Histology of a mediastinal biopsy suggested infective follicular hyperplasia or a peripheral T cell lymphoma. Subsequently, Epstein-Barr virus (EBV) infection was demonstrated in lymphocytes in the biopsy. Later, hepatosplenomegaly and peripheral lymphadenopathy developed. Histology of a cervical lymph node biopsy showed EBV associated diffuse large B cell (non Hodgkin's) lymphoma. PMID- 11872852 TI - An update on pelvic inflammatory disease. AB - A review of the publications on pelvic inflammatory disease (PID) over the past 18 months reveals a number of common themes. This brief article highlights some relevant papers which may be of interest and summarises their main messages. PMID- 11872854 TI - Polymerase chain reaction for diagnosis of genital herpes in a genitourinary medicine clinic. AB - BACKGROUND: Polymerase chain reaction (PCR) has well established advantages over culture for diagnosis of herpes viruses, but its technical complexity has limited its widespread application. However, recent methodological advances have rendered PCR more applicable to routine practice. AIM: To compare automated PCR with viral culture for diagnosis of genital herpes. METHODS: We studied 236 patients presenting with clinical features suggestive of genital herpes at an inner city genitourinary medicine clinic. Two swabs were taken from each patient. Cell culture and typing were performed by standard methods. Automated PCR was performed using the LightCycler instrument and the infecting viral type was determined by restriction endonuclease digestion of amplicons. RESULTS: 109 patients (46%) had a positive test for herpes simplex virus (HSV). In 88, both PCR and culture were positive; in 21 PCR only was positive. With both detection methods, lesion duration and morphology were associated with HSV detection. Compared with culture alone, use of PCR increased sensitivity by 13.3% in specimens from vesicular lesions, by 27.4% from ulcerative lesions, and by 20.0% from crusting lesions. CONCLUSIONS: We advocate adoption of automated PCR as an efficient HSV detection and typing method for diagnosis of genital herpes in routine clinical practice. PCR allowed rapid laboratory confirmation of the diagnosis and increased the overall HSV detection rate by 24%. PMID- 11872855 TI - The accuracy of reported sensitive sexual behaviour in Britain: exploring the extent of change 1990-2000. AB - OBJECTIVES: The 1990-1 British national probability sample survey of sexual attitudes and lifestyles (Natsal 1990) was repeated in 1999-2001 (Natsal 2000) to update population estimates of risk behaviours, and assess change over time. We examine whether changes in prevalence estimates may partly result from changes in measurement accuracy. METHODS: Taking Natsal 2000 (11 161 respondents) and Natsal 1990 (13 765 respondents aged 16-44) we compared the response rate, sample representativeness, reporting of abortion last year (relative to official statistics), and selected attitudes. Among the common birth cohort eligible for both surveys (aged 16-34 Natsal 1990, 26-44 Natsal 2000), we compared reporting of experiences before 1990. RESULTS: The response rate (66.8% Natsal 1990, 65.4% Natsal 2000) and completeness of reporting abortion were unchanged (84% Natsal 1990, 86% Natsal 2000). Attitudes were significantly changed in Natsal 2000 relative to Natsal 1990--for example, increased tolerance of male homosexual sex, OR (95% CI) 2.10 (1.93-2.29) men and 2.95 (2.74 to 3.18) women. In the common birth cohort reporting of heterosexual intercourse before 16 (OR 1.15 (1.02 to 1.29) men, 1.49 (1.31 to 1.69) women), and homosexual experience (OR 1.80 (1.46 to 2.21) men, 2.00 (1.61 to 2.48) women) were significantly increased. CONCLUSIONS: The results are consistent with improved reporting accuracy for some sensitive behaviours in Natsal 2000, in line with greater social tolerance and improved survey methodology. However, the evidence is not conclusive, and may not be generalisable to all such behaviours. The increase found in the reported prevalence of STI risk behaviours between Natsal 1990 and Natsal 2000 is likely to be somewhat overstated. PMID- 11872856 TI - STD in Bangladesh's trucking industry: prevalence and risk factors. AB - OBJECTIVES: This study characterises the prevalence of a broad spectrum of sexually transmitted diseases (STDs) (herpes simplex virus 2, syphilis, chlamydia, gonorrhoea), and examines associations between risk factors and infection in men working in Bangladesh's trucking industry. Given the high risk sexual behaviours of truck drivers and helpers in many contexts, as well as the direct health effects of STDs and their role in facilitating HIV transmission, it is important to understand the prevalence of STDs and associated risk factors in this population. METHODS: A cross sectional study was conducted at Tejgaon truck stand, one of the largest truck stands in Dhaka, the capital city. The study group, comprising 388 truck drivers and helpers, was selected via a two tiered sampling strategy. Of 185 trucking agencies based at the truck stand, 38 agencies were randomly selected, and a mean of 10 subjects (drivers/helpers) were recruited from each agency. Urine and blood samples were collected from subjects after an interview about their lifestyle and a comprehensive physical examination. Gold standard laboratory tests were conducted for the detection of STD. Multiple logistic regression was used to assess associations between infections and potential risk factors. RESULTS: The levels of prevalence of disease were HSV-2 (25.8%), serological syphilis (5.7%), gonorrhoea (2.1%), chlamydia (0.8%). For infection with any bacterial STD (syphilis, gonorrhoea, or chlamydia) the only significant risk factor was having sex with a commercial sex worker in the past year (OR=3.54; CI=1.29-9.72). For HSV-2, truck helpers working primarily on interdistrict routes were significantly more likely to be infected than drivers working on these routes (OR=2.51, CI=1.13--5.55). CONCLUSIONS: The high prevalence of HSV-2, and to a lesser extent syphilis, and the low levels of condom use despite high numbers of casual sexual partners, illustrate the importance of promoting condom use, particularly in commercial sexual encounters, to men in Bangladesh's trucking industry. PMID- 11872857 TI - Pathways to HIV testing and care by black African and white patients in London. AB - OBJECTIVE: To examine factors associated with uptake of HIV clinic services by black African HIV positive people living in London. DESIGN: Questionnaire survey of patients attending study clinic. SETTING: HIV outpatient clinic in south London, UK. SUBJECTS: All HIV positive patients attending the clinic between July 1999 and March 2000. MAIN OUTCOME MEASURES: Use of health services, delay in seeking HIV test, pre-HIV test concerns, delay in uptake of HIV care, barriers to clinic use, disclosure, sources of support. RESULTS: 392 questionnaires were completed. Respondents were 64% white, 26% black African, and 10% from other ethnic groups. Twenty eight per cent of black Africans suspected they were HIV positive before diagnosis (white patients 45% (p<0.01)). Before testing 11% of black Africans had previously attended a genitourinary medicine clinic, 80% had consulted a GP. Twenty per cent of black Africans expressed concern over entitlement to care and where to get an HIV test. The majority of black Africans (66%) received HIV care within 1 month of their diagnosis. They were significantly (p<0.01) less likely than white patients to disclose their HIV status to family and friends. CONCLUSION: This study suggests that although black Africans are a high risk group for HIV infection they generally do not suspect their status. While they may delay testing, their uptake of HIV clinic care and use of statutory and voluntary support services after diagnosis is similar to their white counterparts. However, they lack informal support networks. This study highlights the continuing need for health promotion work among London's African communities, to reduce the stigma surrounding HIV/AIDS and to raise awareness of the benefits of testing. PMID- 11872858 TI - Geographic epidemiology of gonorrhoea and chlamydia on a large military installation: application of a GIS system. AB - OBJECTIVES: The geographic epidemiology of infectious diseases can help in identifying point source outbreaks, elucidating dispersion patterns, and giving direction to control strategies. We sought to establish a geographic information system (GIS) infectious disease surveillance system at a large US military post (Fort Bragg, North Carolina) using STDs as the initial outcome for the model. METHODS: Addresses of incident cases were plotted onto digitised base maps of Fort Bragg (for on-post addresses) and surrounding Cumberland County, NC (for off post addresses) using MAPINFO Version 5. We defined 26 geographic sectors on the installation. Active duty soldiers attending the post preventive medicine clinic were enrolled between July 1998 and June 1999. RESULTS: Gonorrhoea (GC) was diagnosed in 210/2854 (7.4%) and chlamydia (CT) in 445/2860 (15.6%). African American male soldiers were at higher risk for GC (OR = 4.6 (95% CL 3.0 to 7.2)) and chlamydia (OR = 2.0 (1.4 to 2.7)). For women, there were no ethnic differences in gonorrhoea prevalence, but chlamydia was higher in African Americans (OR = 2.0 (1.4-2.7)). Rank and housing type were associated with gonorrhoea and chlamydia in men, but were not significant factors in women. For gonorrhoea, two geographic sectors had prevalences between 14.0%-16.5%, three between 10.3%-13.9%, three between 7.1%-10.2%, and five between 3.0%-7.1%. The geographic distribution demonstrated a core-like pattern where the highest sectors were contiguous and were sectors containing barracks housing lower enlisted grade personnel. In contrast, chlamydia prevalence was narrowly distributed. CONCLUSION: GIS based disease surveillance was easily and rapidly implemented in this setting and should be useful in developing preventive interventions. PMID- 11872859 TI - Sex difference in partner notification: results from three population based surveys in France. AB - OBJECTIVES: To estimate the proportion of individuals in the general population who did not notify their sexual partners at the time of an STD diagnosis, according to the sex of the patient and the type of partner. METHODS: We analysed behaviour at the time of diagnosis of a self reported STD, using data from three large French national population based surveys of adults (ACSF, Barometre Sante) and adolescents (ACSJ). Univariate and multivariate analyses took into account the complex sampling design. RESULTS: In the ACSF, 14% (95% CI: 4% to 24%) of men reported that they had not informed their main sexual partner compared with only 2% (95% CI: 0% to 5%) of women (p = 0.03). This sex difference was independent of the nature of the STD, the patient's age, level of education, and number of partners. Similarly, in the ACSJ, 51% (95% CI: 21% to 81%) of boys reported that they had not talked about this STD with their current sexual partner compared with only 9% (95% CI: 0% to 26%) of girls (p = 0.04). Notification by a sexual partner had led to discovery of the STD more frequently in male subjects than in female subjects, both in adults (32% of men compared with 4% of women (p=0.04)), and adolescents (36% of boys compared with 12% of girls). Most subjects, irrespective of sex, had not informed partners other than their main or current partner: 73% (95% CI: 62% to 84%) of adults and 86% (95% CI: 77% to 95%) of adolescents. CONCLUSIONS: Procedures must be developed urgently to improve the notification of sexual partners, particularly female partners and adolescents, who are unlikely to be tested early without such notification. PMID- 11872860 TI - Surveillance of HIV in the army of the Republic of Cyprus (SHARC); rationale, design, and implementation of an inexpensive system. AB - OBJECTIVES: To design and implement an HIV surveillance system using periodic cross sectional prevalence surveys in National Guard recruits of the Republic of Cyprus. METHODS: HIV infection surveillance used unlinked anonymous screening (UAS) methodology, which tested residual blood originally collected for other purposes. Residual blood from samples collected for ABO blood group typing at intake and samples from blood collected for hepatitis testing at discharge was used. Screening was unlinked and anonymous. RESULTS: The system operated for four semiannual recruitment seasons: summer 1998 to the end of winter 2000. No recruits screened at entry into the ranks tested positive. CONCLUSIONS: This was the first large scale HIV surveillance project in Cyprus. Without nationwide HIV surveys, periodic measurements of prevalence could lead to estimates of HIV incidence and provide insights on temporal changes in HIV infection rates. The prevalence data collected provide useful epidemiological information about the status of the HIV epidemic in this segment of the population in Cyprus. PMID- 11872861 TI - Evaluation of a video based health education strategy to improve sexually transmitted disease partner notification in South Africa. AB - OBJECTIVES: To evaluate the feasibility and impact of a health education intervention promoting partner notification for sexually transmitted diseases (STDs). METHODS: The research setting was a busy public health clinic in a rural district in KwaZulu Natal, South Africa. A before/after quantitative study design was used to measure the effect of an audiovisual presentation of a compelling love drama, posters, and pamphlets. Measures collected from all consenting STD index patients during a 6 week pre-intervention (control) phase were compared with those collected during a 6 week intervention phase. A qualitative evaluation assessed whether the intervention accurately portrayed the intended educational messages. RESULTS: 150 index patients (55% female) were interviewed in the control phase and 185 index patients (64% female) in the intervention phase. The intervention phase showed improvements on several measures of self efficacy about notifying casual partners, such as a belief among index patients that a greater proportion of their casual partners would see the importance of seeking treatment as a result of their notification interaction. The rate of contact cards returned per index patient was 0.27 in the intervention phase, compared with 0.20 in the control phase (95% CI for the rate difference: -0.05, 0.17). The qualitative research found that the intervention was thoroughly enjoyed by patients and clinicians, but a fundamental problem with it was that patients received confused messages about the relation between HIV/AIDS and other STDs. This has potentially negative consequences for partner notification. CONCLUSION: The intervention needs further development, and then could provide a highly acceptable, cost effective model for health education in clinics in developing countries. PMID- 11872863 TI - Mycobacterium xenopi pulmonary infection in an HIV infected patient under highly active antiretroviral treatment. AB - Highly active antiretroviral therapy (HAART) is responsible for a striking reduction in AIDS related morbidity and mortality by partly restoring immune function. However, HAART can also precipitate the development of clinically apparent opportunistic infections in patients with latent infections. We report a case of an HIV infected patient who developed granulomatous nodular and cavitatory lesions of the lungs due to Mycobacterium xenopi as a manifestation of the immune restoration syndrome. PMID- 11872862 TI - Acute onset lactic acidosis and pancreatitis in the third trimester of pregnancy in HIV-1 positive women taking antiretroviral medication. AB - We report the case histories of two HIV-1 positive women in the third trimester of pregnancy who presented with acute lactic acidosis and acute pancreatitis, respectively. One case was fatal for mother and baby. Both women had been stable on regimens containing stavudine and didanosine for at least 2 years before their acute presentations. We speculate on the differential diagnosis, discuss possible reasons for an increased risk of these presentations in pregnant women taking antiretrovirals, and advocate increased vigilance of these women, particularly in the last trimester. PMID- 11872864 TI - Tuberculosis of the cervix: case presentation and a review of the literature. AB - Cervical tuberculosis is uncommon in the developed world. We describe a patient who attended a genitourinary medicine clinic, and who was found to have cervical tuberculosis, in association with HIV infection. PMID- 11872865 TI - Does increased aromatase activity in adipose fibroblasts cause low sexual desire in patients with HIV lipodystrophy? AB - There is evidence from our own unit and other workers that many patients who have lipodystrophy on HAART given for HIV disease also have raised oestrogen levels and complain of low sexual desire. This hypothesis paper discusses a possible pathological mechanism for these changes--an increase in the number of fibroblasts and macrophages present in lipoatrophic areas that could convert testosterone to oestrogen by intracellular aromatisation. This process is known to be enhanced by increased levels of tumour necrosis factor, interleukin 6 (IL 6), and hydroxycorticosteroids present in many patients with HIV lipodystrophy. Treatment options are discussed, including aromatase inhibitors and testosterone. PMID- 11872868 TI - Reproductive morbidity in an Indian urban slum: need for health action. PMID- 11872871 TI - Nevirapine + efavirenz based salvage therapy in heavily pretreated HIV infected patients. PMID- 11872872 TI - Detection of Neisseria gonorrhoeae by PCR using orf1 gene as target. PMID- 11872873 TI - Comparing cost effectiveness of screening women for Chlamydia trachomatis in systematic and opportunistic approaches. PMID- 11872874 TI - Major improvements in cost effectiveness of screening women for Chlamydia trachomatis using pooled urine specimens and high performance testing. PMID- 11872876 TI - Anti-HIV serology in patients with sexual dysphoria in screening test before sex change surgery. PMID- 11872875 TI - Neuropsychiatric reaction induced by clarithromycin. PMID- 11872877 TI - Cost effectiveness analysis of a population based screening programme for asymptomatic Chlamydia trachomatis infection in women. PMID- 11872878 TI - Immune reconstitution eosinophilia due to schistosomiasis. PMID- 11872879 TI - Prevalence of different HIV-1 subtypes in an urban clinic in Madrid. AB - The introduction of non-B HIV-1 variants into Western Europe and North America is of great concern since diagnostic tools and drugs are mainly optimised for targeting HIV-1 subtype B viruses. Madrid is an endemic area for HIV, and either the final destination or transient location for many immigrants from HIV-1 non-B endemic areas (mainly from Africa and South America). From a total of 902 HIV seropositive individuals examined since June 1999, only 27 (3%) were found to be infected with non-B HIV variants. However, up to 71.4% of African immigrants carried non-B viruses. Therefore, HIV subtyping should be requested from all HIV infected African immigrants before considering viral load testing and recommending antiretroviral therapy. PMID- 11872880 TI - Syndromic management of sexually transmitted diseases at primary care level, Mozambique. AB - OBJECTIVE: To assess STD management in primary healthcare facilities where STD clinics and syndromic management guidelines had been introduced. METHODS: In 26 health facilities in selected sites, we observed management of all STD patients presenting in the adult general or STD clinic. We assessed the referral system by analysing patient registers. RESULTS: 408 STD patients (65% in STD and 35% in general clinics) were observed. 70% were women. Women were examined less (26% against 75%, p<0.0001), had laboratory tests ordered more (74% against 45%, p=0.0002), were more often diagnosed syndromically (57% against 38%, p=0.008), and received less advice on condom use (19% against 87%, p<0.001) and contact treatment (47% against 81%, p=0.04). Examination, laboratory requests, diagnosis, and treatment were not significantly different in the STD and general clinic. Health education was better in the STD clinic (condom advice 47% against 8%, p <0.001). Only 41% of referred patients presented to the STD clinic. CONCLUSIONS: The better performance of STD clinics in health education was offset by high referral losses. A proposed integration of STD treatment into general outpatient clinics and better implementation of syndromic management and health education should improve STD case management at primary level in Mozambique. PMID- 11872881 TI - Spontaneous cervical artery dissection: from risk factors toward pathogenesis. PMID- 11872882 TI - Do we really need a better way to give heparin in acute cerebral ischemia? PMID- 11872883 TI - Endothelium-derived hyperpolarizing factor in the brain: a new regulator of cerebral blood flow? PMID- 11872884 TI - Plasma homocysteine concentration, C677T MTHFR genotype, and 844ins68bp CBS genotype in young adults with spontaneous cervical artery dissection and atherothrombotic stroke. AB - BACKGROUND AND PURPOSE: The role of mild hyperhomocysteinemia as a risk factor for cerebral ischemia may depend on stroke subtype. To test this hypothesis, we undertook a prospective case-control study of a group of patients with spontaneous cervical artery dissection (sCAD), a group of patients with atherothrombotic stroke (non-CAD), and a group of control subjects. METHODS: Fasting total plasma homocysteine (tHcy) concentration, C677T MTHFR genotype, and 844ins68bp CBS genotype were determined in 25 patients with sCAD, 31 patients <45 years of age with non-CAD ischemic stroke, and 36 control subjects. Biochemical data in the patient groups were obtained within the first 72 hours of stroke onset. RESULTS: Median tHcy levels were significantly higher in patients with sCAD (13.2 micromol/L; range, 7 to 32.8 micromol/L) compared with control subjects (8.9 micromol/L; range, 5 to 17.3 micromol/L; 95% CI, 1.05 to 1.52; P=0.006). Cases with tHcy concentration above the cutoff level of 12 micromol/L were significantly more represented in the group of patients with sCAD compared with control subjects (64% versus 13.9%; 95% CI, 2.25 to 44.23; P=0.003); a significant association between the MTHFR TT genotype and sCAD was also observed (36% versus 11.1%; 95% CI, 1.10 to 19.23; P=0.045). No significant difference in tHcy levels and in the prevalence of thermolabile MTHFR was found between patients with non-CAD ischemic stroke and control subjects and between patients with sCAD and non-CAD ischemic stroke. The distribution of the 844ins68bp CBS genotype and the prevalence of subjects carrying both the TT MTHFR and 844ins68bp CBS genotypes were not significantly different among the 3 groups. CONCLUSIONS: Our results are consistent with the hypothesis that increased plasma homocysteine levels and the TT MTHFR genotype may represent risk factors for sCAD. In contrast, their role in atherothrombotic strokes remains a contentious issue. PMID- 11872885 TI - Validation of a weight-based nomogram for the use of intravenous heparin in transient ischemic attack or stroke. AB - BACKGROUND AND PURPOSE: Intravenous heparin therapy is often used in patients presenting with transient ischemic attack (TIA) or stroke as either bridging therapy for anticoagulation with warfarin or primary therapy in suspected intracranial arterial dissection, crescendo TIAs, or suspected hypercoagulable states. We attempted to validate the use of a weight-based nomogram for heparin adjusted therapy during hospital admission of patients with TIA or stroke. METHODS: A prospective, single-blinded, randomized, clinical trial was undertaken to compare the use of a specially designed, weight-based heparin nomogram against the traditional method of physician-ordered heparin therapy for patients admitted with TIA or stroke. The trial was not designed to examine the efficacy of heparin therapy but to examine the use of the nomogram for labor requirements, costs of monitoring, safety, length of heparin therapy, and user-friendliness. RESULTS: Pretreatment clinical factors were comparable between those randomized to use of the nomogram (n=101) and to usual care (n=105). Nomogram patients had a significantly lower first activated partial thromboplastin time than nonnomogram patients (60.6 +/- 16.8 versus 69.8 +/- 28.7 seconds). Patients treated by nomogram achieved a therapeutic range of anticoagulation sooner than nonnomogram patients (13.4 +/- 17.0 versus 17.9 +/- 14.1 hours). The fraction of time during which anticoagulation was therapeutic was significantly greater in patients on nomogram therapy (74 +/- 25% versus 67 +/- 26%). Nomogram patients also had significantly fewer supratherapeutic coagulation results, significantly fewer dose adjustment mistakes, significantly fewer calls to house staff regarding anticoagulation, and significantly fewer total complications than nonnomogram patients. The times required for discontinuation of heparin and discharge from hospital were not significantly different. A survey of house staff and nursing staff showed a preference for nomogram use. CONCLUSIONS: The heparin nomogram is a user-friendly method of maintaining heparin infusions and is associated with improved anticoagulation measures, fewer total complications related to heparin therapy, fewer mistakes in heparin dosage adjustment, and decreased labor on the part of house staff and nursing staff. PMID- 11872886 TI - Characteristics of brain arteriovenous malformations with coexisting aneurysms: a comparison of two referral centers. AB - BACKGROUND: Patients harboring a brain arteriovenous malformation (BAVM) often have coexisting arterial aneurysms. Experts have argued about the clinical significance of these aneurysms, which may be important for risk stratification in patient management and clinical trials. We studied the association between coexisting aneurysms and initial presentation with intracranial hemorrhage (ICH) in patients with BAVM evaluated at two tertiary-care centers. METHODS: Demographic and clinical data were collected from a prospective series of patients evaluated for BAVM at the University of California, San Francisco (UCSF; n=82), and Columbia-Presbyterian Medical Center, New York (CPMC; n=254). Using multivariate logistic regression, we examined the independent association between ICH presentation and the presence of a coexisting aneurysm, and compared the association at the two hospitals. RESULTS: Aneurysms were associated with 28 BAVMs at UCSF (34%) and 74 at CPMC (29%; P=0.39). Initial presentation with ICH was associated with the presence of a coexisting aneurysm at CPMC (odds ratio 1.8, 95% confidence interval 1.0 to 3.0, P=0.044). The opposite trend was observed at UCSF (odds ratio 0.4, 95% confidence interval 0.2 to 1.1, P=0.085). We observed an interaction by site involving the association between ICH presentation and aneurysm (P=0.016). CONCLUSION: Although many BAVM characteristics were similar at the referral centers studied, the association between initial presentation with ICH and coexisting aneurysms was not. This heterogeneity between populations undermines the validity of inferences on the role of aneurysms from any single referral series, and emphasizes the complexity in creating BAVM risk-stratification models that incorporate aneurysms. PMID- 11872887 TI - Neurovascular complications of marfan syndrome: a retrospective, hospital-based study. AB - BACKGROUND AND PURPOSE: Small case series have associated Marfan syndrome with cerebral and spinal ischemia or hemorrhage. However, there has been no investigation of the frequency and etiology of neurovascular disorders in a large series of Marfan patients. METHODS: We conducted a retrospective, hospital-based study of all Marfan syndrome patients seen in an 8-year period. Records were reviewed in detail, and clinical characteristics of those with and without a neurovascular diagnosis compared. RESULTS: Of 513 patients, 18 (3.5%) had a neurovascular diagnosis, as follows: transient ischemic attack (11), cerebral infarction (2), spinal cord infarction (2), subdural hematoma (2), and spinal subarachnoid hemorrhage (1). A cardioembolic source was identified in 12 of 13 patients with cerebral ischemia, as follows: prosthetic heart valves (9), mitral valve prolapse (2), and atrial fibrillation (1). Chronic anticoagulant therapy was a likely cause in 2 of 3 patients with hemorrhagic events. Compared with other Marfan syndrome patients, those with neurovascular events were older (39.6 versus 31.7 years, P=0.04) and more likely to be in atrial fibrillation (22.2% versus 3.2%, P=<0.01), to have prosthetic heart valves (61.1% versus 7.7%, P=0.001), and to be taking anticoagulant therapy (72.2% versus 16.1%, P<0.001). Aortic disease, a putative factor in the etiology of neurovascular complications, was present in equal measure in Marfan patients with and without neurovascular complications (78% versus 65%, P=NS). CONCLUSIONS: Neurovascular complications of Marfan syndrome are rare during 8 years of follow-up, and generally are ischemic in nature. A high-risk cardiac source was identified in the majority. A significant association with vascular dissection was not established. PMID- 11872888 TI - Are spontaneous cerebral microemboli consistent in carotid disease? AB - BACKGROUND AND PURPOSE: Transcranial Doppler may be used to detect spontaneous cerebral emboli (SCE), but this information will only identify at-risk patients if these individuals are consistently identified over time. We investigated the consistency of SCE production in patients with symptomatic carotid disease. METHODS: Transcranial Doppler signals from the ipsilateral middle cerebral artery in 25 patients with symptomatic carotid stenosis of >70% were recorded over 1 hour for blind analysis by a panel of trained observers. This was repeated at the same time of day, weekly, for 6 weeks. RESULTS: The number of patients with SCE increased with each week of monitoring until 13 (52%) were positive. The range of the cumulative number of SCE was 1 to 6. Ten (40%) patients were positive for SCE during only 1 monitoring session, and 2 (8%) were positive for SCE during 2 sessions. SCE-positive patients tended to have more recent symptoms. The correlation coefficients between time elapsed since last cerebral symptom and SCE were weak and not significant. CONCLUSIONS: Most, if not all, patients with severe carotid disease will eventually produce SCE. However, the production of an SCE is random, and it is likely that many hours of monitoring are required to determine whether a patient with symptomatic carotid disease is SCE positive. SCE are unlikely to identify at-risk patients but may indicate periods of transiently increased risk in individual patients. PMID- 11872889 TI - Carotid artery intima-media thickness and lacunar versus nonlacunar infarcts. AB - BACKGROUND AND PURPOSE: Increases in the thickness of the intima and media of the carotid artery have been associated with an increased risk of myocardial infarction and stroke in subjects without a history of cardiovascular disease. Lacunar infarcts, one of the most common subtype of ischemic stroke, show unique pathological and clinicoradiological characteristics. The present study examines the relationship between vascular risk factors, including carotid artery intima media thickness (IMT), and lacunar versus nonlacunar infarcts. METHODS: We collected data from patients with acute ischemic stroke admitted to hospital. Patients and 129 control subjects underwent B-mode ultrasonographic measurements of IMT of the common carotid artery. We examined the association of lacunar and nonlacunar infarcts with age, sex, and potential vascular risk factors. RESULTS: Of 292 adult patients with an acute first-ever ischemic stroke, 96 were considered lacunar and 196 were considered nonlacunar strokes. We did not find a significantly different percentage of diabetes, smoking, hypertension, dyslipidemia, myocardial infarction, and previous transient ischemic attack between the 2 groups of patients. The multinomial logistic regression procedure selected carotid artery IMT and atrial fibrillation as the only independent factors able to discriminate between lacunar and nonlacunar patients. IMT values were significantly higher in patients with nonlacunar stroke versus both those with lacunar stroke and control subjects. CONCLUSIONS: The present results indicate the usefulness of noninvasive measurement of IMT with ultrasonic techniques as a diagnostic tool that may help to identify different subtypes of ischemic stroke patients. The noninvasive measurements may have predictive power with respect to lacunar versus nonlacunar infarcts. PMID- 11872890 TI - Venous stasis retinopathy in symptomatic carotid artery occlusion: prevalence, cause, and outcome. AB - BACKGROUND AND PURPOSE: Chronic ocular ischemia is a rare form of ischemia of the eye in patients with carotid artery occlusion (CAO). The early and often asymptomatic stage of chronic ocular ischemia is referred to as venous stasis retinopathy (VSR). The aim of this study was to gain insight into the prevalence, cause, and outcome of VSR in patients with symptomatic CAO. METHODS: In 110 patients with symptomatic CAO, we prospectively investigated the frequency of VSR, the association between the presence of VSR and impaired cerebral blood flow, and the proportion of patients who developed clinically manifest chronic ocular ischemia with ischemia of the anterior eye segment or blindness. RESULTS: At study entry, VSR was found in 32 patients (29%; 95% CI, 21 to 38), particularly in those with symptoms classically associated with a hemodynamic cause, such as limb shaking (relative risk, 2.4; 95% CI, 1.0 to 5.9). Patients with VSR had lower pulsatility indexes in the ophthalmic artery in case of reversed flow, lower cerebral CO2 reactivity, and lower cerebropetal blood flow than patients without VSR. On follow-up (mean, 29 months), clinically manifest chronic ocular ischemia developed in 4 patients (annual rate, 1.5%; 95% CI, 0.4 to 3.8); it tended to occur more often in patients in whom VSR was present at study entry (relative risk, 7.3; 95% CI, 0.8 to 68). CONCLUSIONS: One third of patients with symptomatic CAO has VSR on ophthalmoscopy. VSR is associated with an impaired flow state of the brain. Development of clinically manifest chronic ocular ischemia is rare. PMID- 11872891 TI - Vascular aphasias: main characteristics of patients hospitalized in acute stroke units. AB - BACKGROUND AND PURPOSE: Aphasia is frequent in stroke patients and is associated with poor prognosis. However, characteristics and determinants of vascular aphasias remain controversial. The aim of this study was to evaluate aphasia characteristics at the acute stage in patients admitted to a stroke unit. METHODS: The study was performed in 308 patients consecutively assessed with a standardized aphasia battery. RESULTS: Aphasia was observed in 207 patients; global and nonclassified aphasias accounted for 50% of aphasic syndromes at the acute stage, whereas classic aphasias (Wernicke's, Broca's, transcortical, and subcortical aphasias) were less frequent. Age differed across aphasic syndromes in ischemic stroke patients only; patients with conduction aphasia were younger, and patients with subcortical aphasia were older. Sex did not significantly differ across aphasic syndromes. The presence of a previous stroke was more frequent in nonclassified aphasia. CONCLUSIONS: This study shows (1) that vascular aphasias are frequently severe or nonclassic at the acute stage, a finding explained in part by the presence of a previous stroke; (2) that the age effect is due mainly to its influence on infarct location; and (3) that the main determinant of aphasia characteristics is lesion location. PMID- 11872892 TI - Clinical and imaging findings in cryptogenic stroke patients with and without patent foramen ovale: the PFO-ASA Study. Atrial Septal Aneurysm. AB - BACKGROUND AND PURPOSE: Patent foramen ovale (PFO) has been identified as a potential risk factor for stroke, but the mechanisms of PFO-associated stroke remain unsettled. The aim of our study was to evaluate possible differences in stroke risk factors and stroke patterns between patients with and without PFO that may give clues to the mechanism of PFO-associated stroke. METHODS: This prospective, multicentric study involved 581 young cryptogenic stroke patients. The presence of PFO and atrial septal aneurysm was assessed by transesophageal echocardiography and reviewed independently by 2 experienced sonographers. Clinical, brain, and vascular imaging findings were reviewed by 2 neurologists and 2 neuroradiologists. RESULTS: Of the 581 stroke patients, 267 (45.9%) had PFO. Patients with PFO were younger (OR, 0.95; 95% CI, 0.93 to 0.97) and less likely to have traditional risk factors such as hypertension (OR, 0.49; 95% CI, 0.28 to 0.85), hypercholesterolemia (OR, 0.56; 95% CI, 0.34 to 0.93), or current smoking (OR, 0.67; 95% CI, 0.47 to 0.97). Features suggestive of paradoxical embolism, such as Valsalva-provoking activities or deep vein thrombosis, were not more frequent in patients with PFO. Migraine was more common in patients with PFO (27.3%) than in those without PFO (14.0%). PFO (OR, 1.75; 95% CI, 1.08 to 2.82), particularly when associated with atrial septal aneurysm (OR, 2.71; 95% CI, 1.36 to 5.41), was significantly associated with migraine after adjustment for age and sex. CONCLUSIONS: Differences in stroke risk factors and stroke patterns suggest that different stroke mechanisms occur in patients with and without PFO. PFO is significantly and independently associated with migraine, and this association is even stronger in patients with PFO and atrial septal aneurysm. PMID- 11872893 TI - Microembolus detection in patients with Takayasu's arteritis. AB - BACKGROUND AND PURPOSE: Takayasu's arteritis (TA) is a chronic inflammatory disease of unknown etiology that can affect the aorta and its branches. The cerebral ischemia in TA can be caused by a variety of mechanisms, and the focus of this study is to detect the possible contribution of microembolus in the pathogenesis of stroke. METHODS: Eighteen patients with TA according to the criteria for the classification of TA of the American College of Rheumatology and 100 age-matched healthy controls were studied. Both middle cerebral arteries were monitored by transcranial Doppler (TCD) ultrasound for at least 30 minutes. All patients with TA were followed up for a mean duration of 2.1 months, and recurrent strokes were registered. RESULTS: Microembolic signals (MES) were present in 22% of the patients overall, and the intensity of the MES varied between 9 and 30 dB. Moreover, MES were found in 30% of the patients with higher erythrocyte sedimentation rate. Two (67%) of 3 patients who did not receive any treatment had MES, but only 2 (13%) of 15 patients who received immunosuppressive and anticoagulant therapy before the TCD ultrasonography monitoring had MES. During the follow-up period after MES recording, we did not observe any recurrent stroke. CONCLUSIONS: TCD ultrasonography monitoring can be used as an additional noninvasive procedure to detect microembolus in patients with TA during the acute and chronic phase of the disease. The monitoring of MES may also help in choosing better treatment for the long-term prophylaxis of the disease from acute ischemic stroke, but further large studies are required to justify the efficacy of immunosuppressive treatment in these patients. PMID- 11872894 TI - Predictors of hemorrhagic transformation in patients receiving intra-arterial thrombolysis. AB - BACKGROUND AND PURPOSE: Hemorrhagic transformation (HT) is a major complication of intra-arterial (IA) thrombolytic therapy. Identifying significant predictors of hemorrhage after thrombolysis would be useful in guiding patient selection for IA treatment. METHODS: Data were collected retrospectively on consecutive patients with acute focal cerebral ischemia within the anterior or posterior circulation who were treated with combined intravenous (IV)-IA or pure IA thrombolysis over an 8-year period at the UCLA Medical Center. RESULTS: Eighty nine patients were treated. Median baseline National Institutes of Health Stroke Scale (NIHSS) score was 16, and mean age was 69 years. Twenty-six patients received IA tissue plasminogen activator (tPA) only, 22 received IV-IA tPA, and 41 received IA urokinase only. Asymptomatic HT occurred in 29 patients (33%), minor symptomatic HT (1- to 3-point worsening in NIHSS score) occurred in 10 patients (11%), and major symptomatic HT (> or = 4-point worsening in NIHSS score) occurred in 6 patients (7%). The rate of any HT was similar in patients treated with pure IA thrombolysis (39%) versus combined IV-IA thrombolysis (41%). In pure IA cases, the rate of any HT was 50% with tPA versus 32% with urokinase (P=0.2). Eighty-six percent of the patients with HT versus 39% of the patients without HT were dead or disabled (modified Rankin score >2) at day 7 (P<0.0001). On multivariate analysis, independent predictors of any HT were higher NIHSS score, longer time to recanalization, lower platelet count, and higher glucose level. A model using these variables correctly predicted HT with positive predictive value 70% and overall accuracy 78%. CONCLUSIONS: In this large series of IA thrombolysis, rates of HT were similar to those demonstrated in prior series and clinical trials. Higher NIHSS score, longer time to recanalization, lower platelet count, and higher glucose level were independent predictors of any HT. PMID- 11872895 TI - Abciximab bolus injection does not reduce cerebral ischemic complications of elective carotid artery stenting: a randomized study. AB - BACKGROUND AND PURPOSE: Abciximab has been shown to significantly reduce thromboembolic complications of coronary artery stenting. A prospective, randomized study was performed to test whether abciximab has comparable beneficial effects in carotid artery stenting. METHODS: Seventy-four consecutive patients undergoing elective stenting of the carotid artery were included in the study. Standard antithrombotic medication consisted of aspirin, clopidogrel, and heparin. In addition, half of the patients received an abciximab bolus of 0.25 mg/kg body weight given prophylactically before the intervention. RESULTS: The procedure was successful in all but 1 patient. In patients receiving abciximab, ischemic complications consisted of 4 transient ischemic attacks, 1 minor stroke, 1 nonfatal major stroke, and 1 fatal stroke caused by cerebral hemorrhage. In the control group, 2 transient ischemic attacks and 1 major nonfatal stroke occurred. In summary, the total number of periprocedural ischemic events was 7 (19%) in the abciximab group and 3 (8%) in the control group. Nonischemic complications consisted of 1 inguinal hematoma requiring blood transfusions in each group. CONCLUSIONS: Abciximab bolus given prophylactically before elective carotid artery stenting does not reduce ischemic complications. PMID- 11872896 TI - Fractures after stroke: frequency, types, and associations. AB - BACKGROUND AND PURPOSE: Stroke patients may have an increased risk of fractures because of weak bones or an increased risk of falling. Our goal was to estimate the frequency of fracture after stroke and to identify those at greatest risk. METHODS: This study incorporated 2 complementary strategies: a prospective, single-center, cohort study and an analysis of Scottish routine hospital discharge data. RESULTS: Eighty-eight fractures (30% hip) occurred in 2696 hospital-referred stroke patients. The proportions sustaining any fracture or hip fracture within 2 years were 4% and 1.1%, respectively, 1.4 (95% CI, 0.92 to 2.07) times the rate of hip fracture in the general population (ie, observed number divided by expected number or standardized morbidity ratio). Female sex, older age, low abbreviated mental test score, and prestroke dependence were associated with an increased hip fracture rate. Routine data identified 129 935 acute stroke patients admitted to Scottish hospitals. During 363 447 patient years, 4528 patients had hip fractures, 2.0% had fractures by 1 year, and 10.6% had fractures by 10 years. This is 1.7 times the rate of hip fracture in the general population and 2.3 times that in patients with myocardial infarction. Older patients predictably had the highest rate of poststroke hip fractures but a lower standardized morbidity ratio than younger patients. CONCLUSIONS: Fractures after stroke are probably frequent and serious enough to justify the development of preventive strategies, but the modest event rate would mean that randomized, controlled trials to test these strategies specifically in stroke patients would need to enroll thousands of patients. PMID- 11872897 TI - Old microbleeds are a potential risk factor for cerebral bleeding after ischemic stroke: a gradient-echo T2*-weighted brain MRI study. AB - BACKGROUND AND PURPOSE: T2*-weighted gradient-echo MRI is known to detect old microbleeds (MBs), considered indicative of microangiopathy. MBs might be a potential risk factor for early cerebral bleeding (CB) after ischemic stroke. Therefore, we assessed the impact of MBs on the occurrence of CB after cerebral infarction. METHODS: We included prospectively stroke patients who had documented ischemic damage. The imaging protocol involved baseline CT scan, T2*-weighted gradient-echo MRI, diffusion-weighted imaging, T2-weighted imaging, and magnetic resonance angiography and had to be performed within 24 hours after symptom onset. The assessment of CB with T2*-weighted gradient-echo sequence necessitated a focal area of signal loss either within the ischemic area revealed by diffusion weighted imaging or remote from it. Old MBs were defined on T2*-weighted images as homogeneous rounded areas of signal loss without surrounding edema. CT scan was systematically repeated within the first week to verify CB as diagnosed by the T2* weighted sequence. RESULTS: One hundred patients (mean age, 60 +/- 13 years; range, 19 to 83 years; 58 men, 42 women) met the inclusion criteria. MBs were seen in 20 patients on T2*-weighted imaging. Multivariate logistic regression analysis revealed that age, diabetes, previous use of antithrombotic drugs, evidence of an atherothrombotic source of stroke, and lacunar infarct were significantly associated with MBs (P<0.0001). CB was diagnosed in 26 patients: at the acute stage by T2*-gradient echo sequence in 18 patients and with CT scan performed within the first week in 8 patients. Multivariate logistic regression analysis showed that baseline National Institutes of Health Stroke Scale score, diabetes, and MBs were considered significant and independent predictors of CB (P<0.001). CONCLUSIONS: Although the pathogenesis of CB after ischemic stroke is multifactorial, the increased observation of CB in patients with MBs suggests that the associated vascular vulnerability contributes to CB. PMID- 11872898 TI - Body height is associated with decreased long-term stroke but not coronary heart disease mortality? AB - BACKGROUND AND PURPOSE: There is disagreement whether shorter persons suffer increased rates of coronary heart disease (CHD) or stroke. Potential mechanisms for such associations are not well understood. We used findings from a cohort study, in which 10 000 Israeli men were followed up, to examine the associations between stature and fatal CHD as well as fatal stroke. METHODS: The cohort was composed of 10 059 men aged > or = 40 years who were tenured civil servants or municipal employees. They were followed up for mortality over 23 years (1963 to 1986), for a total of 203 452 person-years of follow-up. We divided men by their height, as measured in the baseline (1963) examinations, into quartiles (< or = 162 cm, 163 to 167 cm, 168 to 171 cm, and > or = 172 cm). RESULTS: During the follow-up period, 1098 men died of CHD, and 364 men died of stroke. Height and weight had been measured for 10 034 men, including all but 1 of the deceased. In contrast to the finding of little variation of CHD death rates between different quartiles of body height, a clear significant pattern of declining stroke mortality (slightly reduced by age adjustment) was observed with increasing body height, with rates of 46, 36, 33, and 29 per 1000 men with increasing height quartiles, respectively (P=0.002 for linear trend). The estimated age-adjusted hazard risk of stroke mortality associated with a 5-cm decrement in height was 1.13 (95% CI 1.04 to 1.22). The respective risk associated with being at the shortest quartile versus the tallest one was 1.54 (95% CI 1.13 to 2.10). Adjustment for socioeconomic status, a predictor of stroke in this cohort, for antihypertensive therapy and for established predictors of stroke (blood pressure, smoking, and diabetes) did not alter these findings. CONCLUSIONS: Height, a potential strong indicator of nutritional status, may be inversely associated with the long-term incidence of fatal stroke in a way that remains to be elucidated. PMID- 11872899 TI - Routine duplex surveillance does not improve the outcome after carotid endarterectomy: a decision and cost utility analysis. AB - BACKGROUND: Doppler ultrasound (duplex) tests are commonly applied after carotid endarterectomy to detect possible recurrent stenosis. The appropriate frequency and the benefits are unknown. We investigated the costs and effects of various follow-up strategies to determine the optimal strategy after carotid endarterectomy. METHODS: Using decision-analytic methods, a Monte Carlo Markov model was constructed. Probabilities and costs were obtained by systematic literature review. From empirical data regarding restenosis, a disease model was constructed to test the effect of various follow-up strategies using duplex testing and angiography. Main outcome measures were quality-adjusted life-years (QALYs), probability of stroke, and costs (for both the Dutch and the American situation). RESULTS: The average quality-adjusted life expectancy for a 66-year old patient was 6.31 years for the symptom-guided strategy (with duplex scanning only being performed in case of symptoms of cerebral ischemia). The mean lifetime costs for this strategy were $5 600 for the US and 4 600 Euro for the Netherlands. The cumulative probability of stroke was 13%. Yearly routine duplex tests up to 5 years after operation resulted in similar QALYs and a similar probability of stroke, but higher costs ($7 300 for the US and 5 600 Euro for The Netherlands situation). No other strategy, including routine duplex surveillance, increased QALYs. When MR instead of conventional angiography was used as confirmatory test, no improvement was observed either. CONCLUSIONS: Routine duplex surveillance does not result in an increase in quality-adjusted life expectancy, but it does increase costs. After successful carotid endarterectomy, a symptom-guided follow-up is an appropriate approach. PMID- 11872900 TI - Functional walk tests in individuals with stroke: relation to perceived exertion and myocardial exertion. AB - BACKGROUND AND PURPOSE: Functional walk tests such as the 6- and 12-Minute Walk Test (ie, 6MWT and 12MWT, respectively) are submaximal measures used to determine functional capacity in individuals with compromised ability. The purpose of this study was to determine the relationship between these walk tests and measures of exertion (perceived and myocardial), in addition to impairment in individuals with stroke. The relationship among the 6MWT, 12MWT, and the more traditionally assessed measure of self-paced gait speed (generally assessed over a short distance, eg, 10 m) was also evaluated. METHODS: Twenty-five community-dwelling individuals with stroke were evaluated for the following: 12MWT distance, 6MWT distance, self-paced gait speed over 8 m, plantarflexion strength, Berg Balance Scale, Ashworth Scale of Spasticity, and Chedoke-McMaster Stroke Assessment. Heart rate (HR), rate-pressure product (RPP), and perceived exertion were assessed during the functional walk tests. Correlational analysis quantified the relationship between gait, impairment measures, and physiological responses during the functional walk tests. RESULTS: HR reached a steady state after 6 minutes and reflected a moderate exercise intensity of 63% of age-predicted maximum HR. The 6MWT, 12MWT, and self-paced gait speed were all highly correlated with one another (r>0.90) and were all also related to the severity of impairments. The functional walk distances did not relate either to perceived exertion or actual exertion (increase in the myocardial oxygen demand as measured by RPP). CONCLUSIONS: Stroke-specific impairments are the major limitations to the distance walked in individuals with stroke. If the functional walk test is used to assess performance of an individual over time (eg, in response to an intervention), we recommend that both exertion (eg, increase in RPP or HR) and distance be measured. PMID- 11872901 TI - Handicap after stroke: how does it relate to disability, perception of recovery, and stroke subtype?: the north North East Melbourne Stroke Incidence Study (NEMESIS). AB - BACKGROUND AND PURPOSE: Knowledge of patterns of handicap after stroke and of the relationship among handicap, disability, perception of recovery, and stroke subtype is limited. The aim of this study was to assess handicap 3 and 12 months after first-ever stroke in a community-based study. METHODS: All strokes occurring in a population of 133 816 people were found and assessed. Patients were classified as having cerebral infarction (CI) or intracerebral hemorrhage (ICH) according to imaging or autopsy findings. Cases of CI were categorized using the Oxfordshire stroke classification. Handicap, disability, and perception of recovery were assessed 3 and 12 months after stroke using the London Handicap Scale, Barthel Index, and the question "Have you made a complete recovery from your stroke?" The association between disability and handicap was examined using Pearson's correlation. Differences in handicap among subtypes of CI were evaluated using one-way ANOVA. RESULTS: There were 264 cases of CI or ICH. Of surviving patients, 113 (59%) were assessed at 3 months and 107 (64%) at 12 months. The domains of handicap most affected were physical independence and occupation. Only half the variance in handicap was due to disability. Of patients without disability, those who claimed complete recovery were less handicapped than those who claimed incomplete recovery. Patients with total anterior circulation infarction were more handicapped at 3 and 12 months than those with other subtypes of CI. CONCLUSIONS: Stroke patients were handicapped across many domains. Handicap is only partly explained by disability. Stroke subtype should be considered in the interpretation of outcome data. PMID- 11872902 TI - Genetic liability in stroke: a long-term follow-up study of Danish twins. AB - BACKGROUND AND PURPOSE: Few studies have assessed the overall importance of genetic factors on stroke risk, and the results have been contradictory. We used a large, population-based twin register and nationwide registries of death and hospitalization with long-term follow-up to estimate the effect of genetic factors on the risk of stroke. METHODS: Through the population-based Danish Twin Register, we identified same-sex twin pairs born in 1870 through 1952 for whom at least 1 twin was recorded under a stroke diagnosis in the Register of Causes of Death or the Danish National Discharge Register. From the day of the first stroke event in each twin pair, the live co-twins were followed up for stroke. In survival analyses, we estimated the age- and sex-adjusted effect of zygosity on the risk of stroke death or hospitalization for stroke. Concordance rates, tetrachoric correlations, and heritability were also assessed. RESULTS: Thirty five of 351 monozygotic pairs (10%) and 34 of 639 dizygotic pairs (5%) were concordant for stroke death. The age- and sex-adjusted relative risk of stroke death in monozygotic compared with dizygotic co-twins was 2.1 (95% CI, 1.3 to 3.3). The probandwise concordance rates were 0.18 (95% CI, 0.14 to 0.22) for monozygotic and 0.10 (95% CI, 0.08 to 0.13) for dizygotic pairs. Thirty-three of 309 monozygotic pairs (11%) and 39 of 560 dizygotic pairs (7%) were concordant for stroke hospitalization or stroke death. The age- and sex-adjusted relative risk of stroke hospitalization or stroke death in monozygotic compared with dizygotic co-twins was 1.5 (95% CI, 0.9 to 2.4). The probandwise concordance rates were 0.19 (95% CI, 0.15 to 0.24) for monozygotic and 0.13 (95% CI, 0.10 to 0.16) for dizygotic pairs. The heritability estimates were 0.32 for the liability to stroke death and 0.17 for the liability to stroke hospitalization or stroke death. CONCLUSIONS: The observed increased risk of stroke death and stroke hospitalization in monozygotic compared with dizygotic co-twins suggests that genetic factors increase the risk of stroke and that the size of this effect is moderate. PMID- 11872903 TI - Inhibitory effect with antisense mitogen-activated protein kinase oligodeoxynucleotide against cerebral vasospasm in rats. AB - BACKGROUND AND PURPOSE: Mitogen-activated protein kinase (MAPK) may be associated with the pathogenesis of cerebral vasospasm after subarachnoid hemorrhage (SAH). This study aimed to clarify the role of MAPK expression and activation during cerebral vasospasm and to evaluate the therapeutic effect on cerebral vasospasm using an antisense MAPK oligodeoxynucleotide (ODN). METHODS: Antisense MAPK, sense MAPK, or scrambled ODN was injected into the rats intracisternally. We used a single-hemorrhage experimental SAH model to assess vasospasm in the basilar arteries at 30 minutes, 1 day, and 2 days after SAH by cross-sectional area measurement and other histological parameters. Immunohistochemistry and Western blot analysis were used to quantify MAPK expression and activation. In addition, a double-hemorrhage rat SAH model was used to test the effect of post-SAH treatment with antisense MAPK ODN. RESULTS: Antisense MAPK therapy significantly inhibited cerebral vasospasm when compared with sense MAPK or scrambled ODN treatment on day 2. The immunohistochemistry and Western blotting performed in the basilar artery of rats that received antisense MAPK ODN demonstrated inhibition of MAPK and phosphorylated MAPK on day 2. In post-SAH treatment study, antisense ODN reduced MAPK and phosphorylated MAPK in the basilar artery and attenuated cerebral vasospasm. CONCLUSIONS: MAPK activation, but not expression, might be implicated with sustained smooth muscle contraction during cerebral vasospasm after SAH. This study suggests that antisense MAPK ODN strategy is an effective treatment against cerebral vasospasm. PMID- 11872904 TI - Protective vasomotor effects of in vivo recombinant endothelial nitric oxide synthase gene expression in a canine model of cerebral vasospasm. AB - BACKGROUND AND PURPOSE: Post-subarachnoid hemorrhage (SAH) cerebral vasospasm is a potentially devastating condition whose pathogenesis involves impaired nitric oxide (NO) bioavailability. We aimed to determine whether recombinant endothelial NO synthase (eNOS) gene expression may protect vasomotor function and prevent vasospasm in a canine experimental SAH model. METHODS: Recombinant adenoviral vectors (5x10(9) plaque-forming units/animal) encoding genes for eNOS (AdeNOS) and beta-galactosidase (AdLacZ) or vehicle were injected into the cerebrospinal fluid (CSF) of dogs on day -1 (ie, 24 hours before the first intra-CSF injection of blood on day 0). Cerebral angiography was performed at day 0 (baseline) and day 7 (immediately before death), and tissues were harvested for additional studies. RESULTS: Western analysis and immunohistochemistry detected recombinant eNOS exclusively in cerebral arteries isolated from AdeNOS-transduced dogs, and in this group of animals CSF NO concentrations were significantly elevated by day 2. Analysis of day 7 versus day 0 cerebral angiograms for each group revealed significant spasm at the basilar artery midpoint in AdLacZ-transduced and nontransduced dogs but not in AdeNOS-transduced dogs. Isometric force recording of basilar arteries isolated from AdeNOS-transduced dogs showed significantly augmented relaxations to bradykinin and reduced contractions to endothelin-1. CONCLUSIONS: Our results suggest that expression of recombinant eNOS in the adventitia of cerebral arteries may contribute toward protection against post-SAH vasospasm. PMID- 11872905 TI - Oxidative stress in rats with heatstroke-induced cerebral ischemia. AB - BACKGROUND AND PURPOSE: Heatstroke is associated with cerebral ischemia as well as increased levels of interleukin-1beta, dopamine, and glutamate in the brain. These factors are known to increase free radical production. This study attempted to ascertain whether an excessive accumulation of cytotoxic free radicals in the brain and oxidative stress can occur during heatstroke. METHODS: Urethane anesthetized rats underwent instrumentation for the measurement of mean arterial pressure, cerebral blood flow, neuronal damage score, and colonic temperature. Rats were exposed to heat stress (ambient temperature, 42 degrees C) until mean arterial pressure and cerebral blood flow began to decrease from their peak levels, which was arbitrarily defined as the onset of heatstroke. Controlled rats were exposed to 24 degrees C. Concentrations of dihydroxybenzoic acid, lipid peroxidation, rate of O2*- generation, superoxide dismutase, and catalase activity of the brain or other vital organs were assessed during heatstroke. RESULTS: The values of mean arterial pressure and cerebral blood flow after heatstroke onset were all significantly lower than those in control rats. However, the values of colonic temperature, dihydroxybenzoic acid levels in the striatum, and neuronal damage score were greater. The extent of lipid peroxidation in the brain and the rate of O2*- generation in the brain, liver, and heart were all greater in rats after heatstroke onset. In contrast, the values of total superoxide dismutase in the brain, liver, and heart and the catalase activity in the brain were lower. CONCLUSIONS: Taken together, these results indicate that hydroxyl radicals mediate cerebral ischemic injury associated with heatstroke. PMID- 11872906 TI - Hypoxic-ischemic injury induces macrophage inflammatory protein-1alpha expression in immature rat brain. AB - BACKGROUND AND PURPOSE: Macrophage inflammatory protein (MIP)-1alpha is a well characterized monocyte chemoattractant; its role in regulating monocyte and microglial recruitment and activation in the injured neonatal brain is unknown. We evaluated the impact of acute hypoxic-ischemic (HI) brain injury on the expression of MIP-1alpha in neonatal rat brain. METHODS: To elicit forebrain ischemic injury, 7-day-old (P7) rats underwent right carotid ligation, followed by 2.5 hours of 8% oxygen exposure. We used an enzyme-linked immunosorbent assay and immunohistochemistry to detect MIP-1alpha; double-labeling immunofluorescence assays were analyzed with confocal microscopy to identify cellular sources of MIP 1alpha. Immunocytochemistry assays were also used to detect 2 MIP-1alpha receptors, CCR1 and CCR5. RESULTS: We found marked increases in tissue concentrations of MIP-1alpha in the HI cerebral hemisphere, peaking from 8 to 72 hours after lesioning. Immunocytochemistry assays revealed that MIP-1alpha was constitutively expressed in physiologically activated microglia; from 8 to 120 hours after lesioning, MIP-1alpha immunoreactive monocytes and microglia accumulated in the lesion territory. In immunoreactive cells, MIP-1alpha was diffusely distributed throughout the cytoplasm at early post-HI time intervals; by 72 hours, MIP-1alpha immunoreactivity was typically concentrated adjacent to the nucleus, a pattern indicative of active MIP-1alpha production. In P7 to P12 brain, many cells expressed MIP-1alpha receptors; both CCR1 and CCR5 immunoreactivity were localized to endothelium and ependyma; CCR1-immunoreactive astrocytes and neurons and CCR5-immunoreactive microglia were also identified. CONCLUSIONS: These data implicate MIP-1alpha as a mediator of the complex and sustained inflammatory response initiated by perinatal HI braininjury. PMID- 11872907 TI - Ca2+ sparks and their function in human cerebral arteries. AB - BACKGROUND AND PURPOSE: Local Ca2+ release events (Ca2+ sparks) caused by the opening of ryanodine-sensitive Ca2+ channels in the sarcoplasmic reticulum have been suggested to oppose constriction in cerebral arteries through the activation of large-conductance Ca2+-activated K+ (BK) channels. We report the first identification and characterization of Ca2+ sparks and associated BK channel currents in smooth muscle cells isolated from human cerebral arteries. METHODS: Membrane currents and intracellular Ca2+ were measured with the use of the patch clamp technique and laser scanning confocal microscopy. RESULTS: Ca2+ sparks with a peak fractional fluorescence change (F/F0) of 2.02 +/- 0.04 and size of 8.2 +/- 0.5 microm2 (n=108) occurred at a frequency of approximately 1 Hz in freshly isolated, cerebral artery myocytes from humans. At a holding potential of -40 mV, the majority of, but not all, Ca2+ sparks (61 of 85 sparks) were associated with transient BK currents. Consistent with a role for Ca2+ sparks in the control of cerebral artery diameter, agents that block Ca2+ sparks (ryanodine) or BK channels (iberiotoxin) were found to contract human cerebral arteries. CONCLUSIONS: This study provides evidence for local Ca2+ signaling in human arterial myocytes and suggests that these events may play an important role in control of cerebral artery diameter in humans. PMID- 11872908 TI - Manganese superoxide dismutase deficiency exacerbates cerebral infarction after focal cerebral ischemia/reperfusion in mice: implications for the production and role of superoxide radicals. AB - BACKGROUND AND PURPOSE: Superoxide anion radicals (O2*-) are implicated in ischemia/reperfusion injury, although a direct relationship has not been elucidated. Recently, a specific method of hydroethidine (HEt) oxidation by O2*- was developed to detect O2*- production in a variety of experimental brain injury models. To clarify the role of O2*- in the mechanism of ischemia/reperfusion, we investigated O2*- production after ischemia/reperfusion and ischemia/reperfusion injury in mutant mice deficient in mitochondrial manganese superoxide dismutase (MnSOD) and in wild-type littermates. METHODS: Ischemia/reperfusion was performed for 60 minutes using intraluminal suture blockade of the middle cerebral artery in the mutant or wild-type mice. We evaluated fluorescent kinetics of HEt or ethidium, the oxidized form of HEt, in brains after an intravenous injection of HEt, followed by measurement of cellular O2*- production using specific HEt oxidation by O2*- before and after ischemia/reperfusion. Furthermore, we compared O2*- production and subsequent infarct volume in the mice using triphenyltetrazolium chloride after ischemia/reperfusion. RESULTS: HEt oxidation to ethidium is primarily a result of mitochondrially produced O2*- under physiological conditions. Cerebral ischemia/reperfusion produced O2*- prominently in neurons shortly after reperfusion, followed by a delayed increase in endothelial cells. A deficiency in MnSOD in mutant mice increased mitochondrial O2*- production and exacerbated cerebral infarction, worsening neurological deficits after ischemia/reperfusion. CONCLUSION: These results suggest that mitochondrial O2*- production may be a critical step underlying the mechanism of ischemia/reperfusion injury and that MnSOD may protect against ongoing oxidative cell death after ischemia/reperfusion. PMID- 11872909 TI - Ultrastructural changes of neuronal mitochondria after transient and permanent cerebral ischemia. AB - BACKGROUND AND PURPOSE: Mitochondrial swelling is one of the most striking and initial ultrastructural changes after acute brain ischemia. The purpose of the present study was to examine the role of reperfusion of the cerebral cortex after transient focal cerebral ischemia on neuronal mitochondrial damage. METHODS: Male Sprague-Dawley rats (n=16) were subjected to either temporary or permanent occlusion of the middle cerebral artery and bilateral carotid arteries. Three experimental conditions were compared: group I, permanent ischemia (3, 5, and 24 hours); group II, transient ischemia (2, 24 hours of reperfusion); and sham surgery. Anesthetized rats were killed by cardiac perfusion, and brain tissue was removed ipsilaterally and contralaterally from the ischemic core section of the frontoparietal cortex. Fixed tissue was prepared for electron microscopic examination, and electron microscopic thin sections of random neurons were photographed. Perinuclear neuronal mitochondria were analyzed in a blinded manner for qualitative ultrastructural changes (compared with sham control) by 2 independent investigators using an objective grading system. RESULTS: Cortical neuronal mitochondria exposed to severe ischemic/reperfusion conditions demonstrated dramatic signs of injury in the form of condensation, increased matrix density, and deposits of electron-dense material followed by disintegration by 24 hours. In contrast, mitochondria exposed to an equivalent time of permanent ischemia demonstrated increasing loss of matrix density with pronounced swelling followed by retention of their shape by 24 hours. CONCLUSIONS: Neuronal mitochondria undergoing transient versus permanent ischemia exhibit significantly different patterns of injury. Structural damage to neuronal mitochondria of the neocortex occurs more acutely and to a greater extent during the reperfusion phase in comparison to ischemic conditions alone. Further research is in progress to delineate the role of oxygen free radical production in the observed mitochondrial damage during postischemic reoxygenation. PMID- 11872910 TI - New insights into brain damage in stroke-prone rats: a nuclear magnetic imaging study. AB - BACKGROUND AND PURPOSE: The spontaneously hypertensive stroke-prone rat (SHRSP) is an animal model for a complex form of cerebrovascular pathology. MRI provides an efficient and noninvasive tool for studying the time course of brain damage. The aim of this study was to gain new insights into the pathological phenomena responsible for the occurrence of brain injury in SHRSP with the use of the apparent diffusion coefficient of water (ADC), one of the most efficient MRI parameters for detecting brain abnormalities. To this end, the pattern of ADC variation observed in SHRSP was compared with that of focal ischemia induced in both SHRSP and Sprague-Dawley rats. METHODS: Four groups of animals were studied: SHRSP developing spontaneous brain lesions fed with a salt-loaded (n=15, group 1) or standard diet (n=3, group 2) and Sprague-Dawley rats (n=8, group 3) and SHRSP (n=8, group 4) with permanent middle cerebral artery occlusion. ADC maps and T2 weighted images of brains were performed by MRI. After the rats were killed, the brains were removed and histologically processed. RESULTS: There was no decrease in ADC during spontaneous stroke in the SHRSP fed with a normal or salt-enriched diet, while both the SHRSP and Sprague-Dawley rats with middle cerebral artery occlusion showed a marked decrease that lasted for 24 to 48 hours. CONCLUSIONS: Cerebral ischemia cannot be considered a major factor in the onset of spontaneous brain lesions in SHRSP, which show only vasogenic edema after the beginning of the damage with no evidence of metabolic impairment. PMID- 11872911 TI - Involvement of matrix metalloproteinase in thrombolysis-associated hemorrhagic transformation after embolic focal ischemia in rats. AB - BACKGROUND AND PURPOSE: Thrombolytic therapy with tissue plasminogen activator (tPA) for acute ischemic stroke remains complicated by risks of hemorrhagic transformation. In this study we used a previously established quantitative rat model of tPA-associated hemorrhage to test the hypothesis that matrix metalloproteinases (MMPs) are involved. METHODS: Spontaneously hypertensive rats were subjected to embolic focal ischemia by placing homologous blood clots into the middle cerebral artery. Three groups of rats were studied: (1) untreated controls that received saline at 6 hours after ischemia; (2) rats that received tPA alone (10 mg/kg at 6 hours after ischemia); and (3) rats that received tPA plus the broad-spectrum MMP inhibitor BB-94 (50 mg/kg of BB-94 before ischemia and at 3 and 6 hours after ischemia plus tPA at 6 hours). Gelatin zymography was used to quantify MMP levels. A hemoglobin spectrophotometry method was used to quantify cerebral hemorrhage. Ischemic lesions were measured at 24 hours with tetrazolium staining. RESULTS: At 6, 12, and 24 hours, pro-MMP-9 and cleaved MMP 9 were upregulated in ischemic brain. At 12 hours, tPA-treated rats showed significantly higher levels of pro-MMP-9 and cleaved MMP-9 than untreated controls. By 24 hours, all rats showed evidence of hemorrhagic transformation in the ischemic territory. Rats treated with BB-94 and tPA showed significantly reduced hemorrhage volumes compared with those that received tPA alone. There was no effect on infarct size. CONCLUSIONS: These results indicate that (1) tPA treatment increases levels of MMP-9 after embolic focal cerebral ischemia, (2) MMPs are involved in the mechanism of tPA-associated hemorrhage, and (3) combination therapies with MMP inhibitors may be useful for decreasing the risk and severity of this dreaded complication of thrombolytic therapy. PMID- 11872912 TI - Dependence of early cerebral reperfusion and long-term outcome on resuscitation efficiency after cardiac arrest in rats. AB - BACKGROUND AND PURPOSE: While it is well known that longer duration of cardiac arrest (CA) is often associated with poorer long-term outcome, the influence of resuscitation efficacy on postischemia recovery is less clear. The objective of the present study is to investigate whether an inadequate and prolonged resuscitation after a shorter CA can lead to worse long-term outcomes than an effective resuscitation after a longer CA, provided that the total time from the onset of CA to the return of spontaneous circulation is comparable. METHODS: Thirty-eight rats were randomized into 2 groups with nominal 9 minutes (group 1) and 15 minutes (group 2) of normothermic asphyxial CA. Each group was further divided into 2 subgroups on the basis of the duration of resuscitation efforts (labeled as S and L for short and long, respectively). Thus, the asphyxia and nominal resuscitation times were 8 and 1 minute, respectively, for group 1S, 5 and 4 minutes for group 1L, 14 and 1 minute for group 2S, and 11 and 4 minutes for group 2L. Cerebral perfusion was measured continuously at the dorsal hippocampus level before, during, and after the CA, with the use of the arterial spin labeling MRI technique. The survival time, histological damage, and neurological deficit were evaluated 5 days after resuscitation. RESULTS: Groups 1S and 1L had nearly the same duration of CA (9.02 +/- 0.17 minutes, n=6 versus 8.58 +/- 0.80 minutes, n=6). The same is true for groups 2S and 2L (15.51 +/- 0.59 minutes, n=11 versus 15.65 +/- 1.25 minutes, n=15). Despite longer asphyxia, shorter and more effective resuscitation was associated with significantly improved long-term outcomes and higher cerebral perfusion at the early stage of reperfusion. CONCLUSIONS: Effective resuscitation increased early reperfusion and improved survival after CA. The clinical implication is that inadequate and prolonged resuscitation may have detrimental effects on the recovery of CA patients. PMID- 11872913 TI - Role of endothelial nitric oxide and smooth muscle potassium channels in cerebral arteriolar dilation in response to acidosis. AB - BACKGROUND AND PURPOSE: Potassium channels or nitric oxide or both are major mediators of acidosis-induced dilation in the cerebral circulation. However, these contributions depend on a variety of factors such as species and vessel location. The present study was designed to clarify whether potassium channels and endothelial nitric oxide are involved in acidosis-induced dilation of isolated rat cerebral arterioles. METHODS: Cerebral arterioles were cannulated and monitored with an inverted microscope. Acidosis (pH 6.8 to 7.4) produced by adding hydrogen ions mediated dilation of the cerebral arterioles in a concentration-dependent manner. The role of nitric oxide and potassium channels in response to acidosis was examined with several specific inhibitors and endothelial damage. RESULTS: The dilation was significantly inhibited by potassium chloride (30 mmol/L) and glibenclamide (3 micromol/L; ATP-sensitive potassium channel inhibitor). We found that 30 micromol/L BaCl2 (concentration dependent potassium channel inhibitor) also affected the dilation; however, an additional treatment of 3 micromol/L glibenclamide did not produce further inhibition. Tetraethylammonium ion (1 mmol/L; calcium-activated potassium channel inhibitor) and 4-aminopyridine (100 micromol/L; voltage-dependent potassium channel inhibitor) as well as ouabain (10 micromol/L; Na-K ATPase inhibitor) and N-methylsulphonyl-6-(2-proparglyloxyphenyl) hexanamide (1 micromol/L; cytochrome P450 epoxygenase inhibitor) did not alter acidotic dilation. N(omega)-Monomethyl L-arginine (10 micromol/L) and N(omega)-nitro-L-arginine (10 micromol/L) as nitric oxide synthase inhibitor blunted the dilation. Furthermore, the dilation was significantly attenuated after the endothelial impairment. Additional treatment with glibenclamide (3 micromol/L) further reduced the dilation in response to acidosis. CONCLUSIONS: Endothelial nitric oxide and smooth muscle ATP sensitive potassium channels contribute to acidosis-induced dilation of rat cerebral arterioles. Endothelial damage caused by pathological conditions such as subarachnoid hemorrhage or traumatic brain injury may contribute to reduced blood flow despite injury-induced cerebral acidosis. PMID- 11872914 TI - Role of the endothelial lining in persistence of residual lesions and growth of recurrences after endovascular treatment of experimental aneurysms. AB - BACKGROUND AND PURPOSE: We sought to investigate the role of the endothelial lining in aneurysmal persistence and recurrence after endovascular treatment of aneurysms. METHODS: Bilateral venous pouch canine carotid aneurysms were studied by angiography and pathology 1, 2, and 3 weeks after intraoperative collagen sponge embolization, coated or not coated with fibrinogen (n=15). In 12 dogs the endothelial lining of one aneurysm was removed before embolization, and results were compared with a control aneurysm after bilateral sponge embolization. In 4 animals embolization with a sponge covered with endothelium was compared with treatment with a sponge covered with adventitia. In 4 animals an inverted venous pouch aneurysm embolized with an adventitia-covered sponge was compared with a normal aneurysm embolized with an endothelialized sponge. In 3 animals inverted venous pouch aneurysms were embolized with a sponge covered or not covered with endothelium. Control aneurysms (n=3 or 4 each) included untreated normal, inverted, and de-endothelialized venous pouch aneurysms. Angiographic results at 3 weeks were compared by Wilcoxon's test. RESULTS: Endothelialization of the clot that forms on the sponge was complete at 1 week, forming clefts that developed into recurrences. Reversed or de-endothelialized aneurysms spontaneously thrombosed, while normal venous pouch aneurysms remained patent for at least 3 weeks. The addition of fibrinogen, endothelium, or adventitia to the sponges did not prevent recurrences, which occurred routinely after embolization of endothelialized aneurysm. De-endothelialization of the aneurysmal wall improved angiographic results at 3 weeks (P=0.02), while reversing the venous pouch before embolization led to complete healing (P=0.003). CONCLUSIONS: The endothelial lining is essential to the persistence of residual lesions. Early endothelial invasion of the clot leads to recanalization and recurrences after embolization of aneurysms. This observation provides new opportunities to improve results of endovascular treatment of aneurysms. PMID- 11872915 TI - Emergent use of anticoagulation for treatment of patients with ischemic stroke. AB - BACKGROUND: Several clinical trials have tested the potential utility of emergent anticoagulation for acute ischemic stroke. SUMMARY OF REVIEW: Rather than performing a meta-analysis that combines the data from several trials, this review focuses on individual studies. Although these trials do have inherent limitations, they demonstrate that emergent use of an anticoagulant is associated with a modest but significantly increased risk of hemorrhagic transformation of the ischemic stroke or serious nonneurological bleeding. The trials do not demonstrate a benefit from emergent anticoagulation in improving outcome, reducing mortality, and preventing early recurrent stroke. CONCLUSIONS: These results suggest that most patients with acute stroke should not be treated with unfractionated heparin or other rapidly acting anticoagulants after stroke. Prevention of deep vein thrombosis and pulmonary embolism among bedridden patients is the only established indication for early anticoagulation after acute ischemic stroke. PMID- 11872916 TI - Stroke prevention therapy beyond antithrombotics: unifying mechanisms in ischemic stroke pathogenesis and implications for therapy: an invited review. AB - BACKGROUND: It is estimated that about half of cardiovascular disease risk is explained by conventional risk factors. The realization that atherosclerosis is an inflammatory disease has led to a search for new stroke and cardiovascular disease risk factors and treatments. As such, the vulnerable atherosclerotic plaque has become the main focus for new medical strategies for plaque stabilization and stroke prevention. SUMMARY OF REVIEW: In this invited review, I discuss inflammation as a possible risk factor for stroke, unifying mechanisms in ischemic stroke pathogenesis, and new avenues for stroke prevention---statin agents, angiotensin-converting enzyme inhibitors, and vitamins. These new stroke prevention therapies may help to reduce inflammation, serve to stabilize the atherosclerotic plaque, or act by other protective mechanisms. CONCLUSION: Beyond the traditional antithrombotic agents, statin agents, angiotensin-converting enzyme inhibitors, and vitamins may prove to be important additions to our armamentarium for stroke prevention. PMID- 11872917 TI - The unusually shaped bifrontal hematoma. PMID- 11872918 TI - Re: Feasibility and safety of moderate hypothermia after massive hemispheric infarction. PMID- 11872919 TI - Re: Crossed nonaphasia in a dextral with left hemispheric lesions: a functional magnetic resonance imaging study of mirrored brain organization. PMID- 11872920 TI - An unjustified return to the past. PMID- 11872923 TI - Measurement of the speed of X-rays. AB - X-ray pulses from the Advanced Photon Source at Argonne National Laboratory were used to measure the speed of X-rays in the energy range between 21 and 60 keV. An LiNbO(3)-based 0.58 GHz surface acoustic wave device served as a temporal analyzer in the stroboscopic time-resolved diffraction measurements. By synchronizing the surface acoustic wave excitation and periodic X-ray illumination of the LiNbO(3) crystal, the temporal modifications in the LiNbO(3) diffraction profiles could be followed and the time points of X-ray arrivals at the analyzer position for different analyzer to storage ring distances determined. The speed of the X-rays was determined as the ratio of measured spacings and corresponding delay time intervals. Within the experimental error bars, the obtained X-ray velocities converged to the tabulated constant for the speed of light in a vacuum. PMID- 11872921 TI - Age-adjusted stroke incidence increase: could angiotensin AT1 receptor antagonists enhance stroke prevention? PMID- 11872924 TI - Quantitative investigations of supported metal catalysts by ASAXS. AB - Limits and potentiality of anomalous small-angle X-ray scattering for the investigation of supported metal catalysts are discussed. The different sources of statistical errors are illustrated using two catalysts containing metals with absorption edges at very different energies (Au and Pd). Optimized experimental conditions and measuring strategy are proposed, and data-evaluation methods for obtaining quantitative reliable results are suggested. By this method, Au content as low as 0.2 wt% and Pd content of 3 wt% could be investigated with success. The detection limits for palladium are higher, mainly due to its smaller electronic contrast. PMID- 11872925 TI - Synchrotron fibre diffraction identifies and locates foetal collagenous breast tissue associated with breast carcinoma. AB - Synchrotron fibre diffraction studies of collagenous breast tissue have revealed clear and consistent differences in the diffraction intensity patterns between samples taken from patients with breast carcinoma and those taken from controls. These changes, prelusive to carcinoma, are related to changes in and breakdown of the molecular structure of the collagen type IV support collagen adjacent to the type I, type III and type V collagen fibrils. In order to locate the positions and sequence of the different molecular arrangements, multiple samples were taken from a number of patients. The first of these was taken as near to the tumour site as was possible. Subsequent samples were taken at intervals along ducts leading away from the tumour. The results reveal that the collagen distal from the tumour is similar to that of the controls and this was taken as the standard molecular structure of breast tissue. It usually contained some fat. For normal samples taken closer to the tumour site, the wide diffraction ring associated with fat disappears leaving only the fibrillar pattern. Moving closer to the tumour a 'foetal-like tissue' was observed. The tissue immediately adjacent to the tumour was found to have the same molecular structure as that of foetal tissue. PMID- 11872926 TI - Fast residual stress mapping using energy-dispersive synchrotron X-ray diffraction on station 16.3 at the SRS. AB - Synchrotron energy-dispersive X-ray diffraction experiments on station 16.3 at the SRS for residual strain mapping are reported. A white beam with an energy discriminating detector allows measurements to be made through 3 mm Al, Ti, Fe and Cu alloys with acquisition times of approximately 30 s per 0.3 mm(3) sampling volume. The collected profiles were analysed using single-peak fitting and whole pattern Pawley refinement, and produced strain accuracy better than 10(-4). This configuration is therefore highly efficient for fast strain mapping in thin components using a second-generation synchrotron source. PMID- 11872927 TI - Short X-ray pulses in a Laue-case crystal. AB - The short X-ray pulses coming out of a SASE FEL (self-amplified stimulated emission free-electron laser) have stimulated a closer inspection of the response of a crystal reflection to them. After a short collection of formulae taken from the dynamical theory of X-ray diffraction, the response to a delta-pulse reflected by a Laue-case monochromator crystal is investigated. In contrast to the already discussed Bragg-case monochromator, a two-dimensional analysis is required. PMID- 11872928 TI - New opportunities in trace elements structural characterization: high-energy X ray absorption near-edge structure spectroscopy. AB - Garnets in lower crustal mafic and ultramafic rocks usually contain rare-earth elements (REE) in trace concentrations. Direct characterization of REE at trace levels in natural garnets is not available in the literature because of the difficulty of obtaining structural information by means of conventional diffraction methods. Here, the characterization of Nd at trace levels (176-1029 p.p.m.) in a set of natural garnets performed by means of Nd K-edge X-ray absorption near-edge structure spectroscopy is presented, showing the capability of high-energy XANES for REE in trace structural determinations. PMID- 11872929 TI - Reconstruction of magnetization density in two-dimensional samples from soft X ray speckle patterns using the multiple-wavelength anomalous diffraction method. AB - A non-destructive technique for imaging magnetic domains in thin films and two dimensional magnetic structures using coherent soft X-ray scattering and the multiple-wavelength anomalous diffraction method (MAD) is proposed. The method exploits the strong energy dependence in the magnetic scattering amplitude for 3d transition metals near the L(2,3) absorption edges and 4f elements near the M(4,5) absorption edges. The phase information required in the reconstruction algorithm is derived from the interference between the charge and magnetic scattering amplitudes. Magnetic speckle patterns from the magnetic domain distribution in an artificially defined Fe thin film are used to demonstrate this reconstruction algorithm. Circularly and linearly polarized incident light are examined separately to investigate the effect of polarization on the capability of the method. PMID- 11872930 TI - Optical switching of X-rays using laser-induced lattice expansion. AB - The optical switching of X-rays using laser-induced crystal lattice expansion is described. Irradiation of a gallium arsenide (GaAs) crystal using picosecond laser pulses shifts the Bragg angle through the lattice expansion with a response time of a few hundred picoseconds. A single pulse was extracted from the synchrotron radiation pulse train using a double-crystal arrangement of GaAs, in which the two crystals were irradiated by way of two successive laser pulses with an appropriate time delay. PMID- 11872931 TI - Fast multigrid fluorescent ion chamber with 0.1 ms time response. AB - A fast multigrid ion chamber for the detection of fluorescent X-rays has been developed. The structure of 17 grids with close separation was employed to maximize the time response as well as to give sufficient detection efficiency. The measured rise/fall response time to cyclic X-rays was shorter than that of an existing three-grid ion chamber by more than one order of magnitude. A 0.13 ms time response was obtained at the 500 V applied voltage, where the detector can stably operate without any discharge. The available frequency range is as high as 1 kHz with a practical amplitude response. PMID- 11872932 TI - Changes in the perception of biological X-ray absorption spectroscopy between ICBIC 1 and ICBIC 10/BIOXAS 2001: how far have we travelled? AB - The proceedings of BIOXAS 2001 (Siena, Italy, 2001) and both current and planned activities in the field of biological X-ray absorption spectroscopy are discussed against the perspective of changes in the perception of the technique since ICBIC 1 (Florence, Italy, 1983). PMID- 11872940 TI - Sexually dimorphic effect of glutamate treatment on cell cycle arrestment of astrocytes from the preoptic area of neonatal rats. AB - Neurotoxicological studies have indicated that L-glutamate exhibits more pronounced effects on the preoptic area (POA) neurons of male rats than on those of females in the neonatal period. However, no information has previously been available as to whether or not such sexual dimorphism also exists for the effects of glutamate on astrocytes from POA. The present paper reports the differential effects of L-glutamate on astrocytes isolated from POA of neonatal male and female rats. The proliferation of astrocytes was measured by methods of cell count and cell cycle analysis. In addition, the activity of Ca(2+)/calmodulin dependent protein kinase II (CaM kinase II) was assayed to understand its role in the glutamate-induced disturbance of the cell cycle progression of astrocytes. The results revealed that L-glutamate, at doses of 0.5 and 1.0 mM, inhibited the proliferation of astrocytes derived from male rats more severely than those derived from females. The L-glutamate treatment blocked the cell cycle progression and caused an accumulation of cells in the S phase. The activity of CaM kinase II declined more markedly in astrocytes derived from male rats than in those from females after glutamate treatment. These findings suggest that the proliferation of astrocytes derived from POA of neonatal rats can be inhibited in a sexually dimorphic manner by L-glutamate, possibly through blocking the cell cycle progression and partially related to the inactivation of the CaM kinase II. PMID- 11872939 TI - Study of the role of the low-affinity neurotrophin receptor p75 in naturally occurring cell death during development of the rat retina. AB - The aim of this study was to examine the tempo-spatial expression of low-affinity neurotrophin receptor p75, or p75(NTR), and its role in the induction of retinal ganglion cell (RGC) apoptosis in the rat retina during development. The cellular distribution of p75 in the retina was demonstrated with immunohistochemistry and double-immunofluorescent staining. Apoptosis in the developing rat retina was detected by DNA gel electrophoresis, and the number of RGCs undergoing apoptosis was estimated by terminal deoxyribonucleotidyl-mediated dUTP-digoxigenin nick end labeling (TUNEL). To localize p75 on apoptotic RGCs, p75 immunofluorescence and TUNEL fluorescent staining was performed on sections with Fluoro-Gold-prelabeled RGCs. p75 immunoreactivities were not detected either on the RGCs or TUNEL positive cells, whereas Muller cell processes were p75 immunopositive. Thus, it was most unlikely that p75 induced apoptosis of RGCs in the rat retina. PMID- 11872941 TI - Differential response of immature and mature neurons to hypoxia in rat mesencephalic cultures. AB - The effect of hypoxia on immature and mature mesencephalic neurons was studied in in vitro rat cerebral cell cultures on different days. In immature cultures (6-8 days in vitro), exposure to 24 h of hypoxia (10-20 mm Hg pO(2) in the culture medium) did not change the number of neuron-specific enolase (NSE)-immunoreactive (IR) (NSE-IR) neurons but increased the number of tyrosine hydroxylase (TH)-IR (TH-IR) cells, which might be attributed to transient induction of TH. In mature cultures (13-15 days in vitro), 16 h of hypoxia induced a considerable loss of both NSE- and TH-IR cells. A decrease in the number of TH-IR cells 6 and 24 h after hypoxia was more pronounced than that of NSE-IR cells; however, their numbers equalized 48 h after hypoxia, suggesting similar hypoxic vulnerability of dopaminergic and nondopaminergic neurons in mature mesencephalic cultures. In immature cultures, hypoxia slightly stimulated both apoptosis and necrosis, while in mature cultures, it dramatically increased the number of solely necrotic cells. PMID- 11872942 TI - Greater resistance and lower contribution of free radicals to hypoxic neurotoxicity in immature rat brain compared to adult brain as revealed by dynamic changes in glucose metabolism. AB - Seven-day-old rat brain slices were incubated at 36C in oxygenated Krebs-Ringer solution containing [(18)F]2-fluoro-2-deoxy-D-glucose ([(18)F]FDG), and serial two-dimensional time-resolved images of [(18)F]FDG uptake by the slices were obtained. The Gjedde-Patlak graphical method was applied to the image data, and the duration limit of hypoxia loading that allowed recovery of the fractional rate constant (k3*) of [(18)F]FDG (proportional to the cerebral glucose metabolic rate) after hypoxia loading to the unloaded control level was 50 min, and MK-801 as an N-methyl-D-aspartate antagonist had neuroprotective effects, but PBN as a free radical scavenger was ineffective. In our previous study in adult (7-week old) rat brains [Murata et al., Exp Neurol 2000, 164:269-279], the limit of the hypoxia loading time was 20 min, and both MK-801 and PBN were effective. In the immature rat brains, the ratio of aerobic glucose metabolism to the total glucose metabolism was low compared with the adult rat brains, suggesting only a slight involvement of free radicals in hypoxic neurotoxicity. These data suggest that the higher resistance of immature brains to hypoxia compared to that of adult brains is attributable to a lower involvement of free radicals due to a lower aerobic glucose metabolic rate. PMID- 11872943 TI - Gap junction-mediated bidirectional signaling between human fetal hippocampal neurons and astrocytes. AB - Gap junctions are clusters of intercellular channels that connect the interiors of coupled cells. In the brain, gap junctions function as electrotonic synapses between neurons and as pathways for the exchange of metabolites and second messenger molecules between glial cells. Astrocytes, the most abundant glial cell type coupled by gap junctions, are intimately involved in the active control of neuronal activity including synaptic transmission and plasticity. Previous studies have suggested that astrocytic-neuronal signaling may involve gap junction-mediated intercellular connections; this issue remains unresolved. In this study, we demonstrate that second-trimester human fetal hippocampal neurons and astrocytes in culture are coupled by gap junctions bidirectionally; we show that human fetal neurons and astrocytes express both the same and different connexin subtypes. The formation of functional homotypic and heterotypic gap junction channels between neurons and astrocytes may add versatility to the signaling between these cell types during human hippocampal ontogeny; disruption of such signaling may contribute to CNS dysfunction during pregnancy. PMID- 11872944 TI - Dopamine induces phenotypic differentiation or apoptosis in a dose-dependent fashion: involvement of the dopamine transporter and p53. AB - The effect of dopamine on the growth, phenotypes (morphological and biochemical) and programmed cell death (apoptosis) of the human neuronal NMB cell line was examined. Exposure to 20-50 microM of dopamine decreased cell growth, induced an apparent differentiated cell morphology and increased (3)H-dopamine uptake. At higher concentrations (100-300 microM) dopamine was neurotoxic and induced apoptosis, as reported previously. The observed effects of both low and high doses of dopamine were blocked by cocaine, which suggested involvement of dopamine transporters. Indeed, several experiments demonstrated the relationship between dopamine uptake of cells and their vulnerability to the toxic effect of dopamine. High concentrations of dopamine, which induced apoptosis, also increased p53 levels, detected by RT-PCR analysis and immunoblotting, whereas lower dopamine concentrations, which induced a differentiated phenotype, did not increase p53 immunoblotting. Dibutyryl-cAMP and dimethyl sulfoxide, which induced differentiation but not apoptosis of the NMB cells, did not increase p53 expression. These findings provide an insight into the role of dopamine, dopamine transporters and p53 in the differentiation and apoptosis of dopaminergic neurons, which will further our understanding of neuronal development and neurodegenerative diseases. PMID- 11872945 TI - Testosterone regulates terminal Schwann cell number and junctional size during developmental synapse elimination. AB - Previous work has shown that exposure to exogenous testosterone during synapse elimination permanently stabilizes synapses that would normally be lost in the androgen-sensitive levator ani (LA) muscle, indicating that testosterone is a potent stabilizing factor for developing LA synapses. Terminal Schwann cells (TSCs), which cap the neuromuscular junction, have also been implicated in the control of synaptic stability and may play a decisive role in the selective stabilization of synapses during synapse elimination. In this study, we begin to investigate the possible role of TSCs in the effect of testosterone on synapse elimination by determining whether testosterone influences their number. As the number of TSCs generally correlates with the size of endplates, we also measured endplate size. Male rats were castrated or sham gonadectomized at postnatal day (P) 7 and given capsules containing either testosterone or nothing. Three weeks later (P27-28), LA neuromuscular junctions (NMJs) were stained using immune and nonimmune markers. As expected, testosterone treatment during synapse elimination maintained synapses that would ordinarily be eliminated. In addition, we find that the size of LA endplates and the number of TSCs per LA junction were also increased by the testosterone treatment. However, testosterone significantly increased the number of TSCs on both singly and multiply innervated fibers, indicating that the effect of testosterone on the number of TSCs is not specific to its effect on synapse maintenance. Rather the testosterone-induced increase in the number of TSCs appears related to the size of LA NMJs, a relationship that has precedence. PMID- 11872946 TI - Identification of genes in the oligodendrocyte lineage through the analysis of conditionally immortalized cell lines. AB - The mouse oligodendrocyte cell lines, N19 and N20.1, were used as sources of potential stage-specific RNA in order to construct a subtraction library enriched in cDNAs expressed early in the oligodendrocyte (OL) lineage. From this library, 23 clones were examined and three were examined in most detail. The mRNAs of the three library clones were preferentially expressed in the N19 (progenitor) compared to the N20.1 (immature) OL line. One of these corresponded to the intermediate filament protein cytokeratin K19, which has not been reported to be expressed in OLs previously. Another was identified as the mouse homolog of T cadherin, previously reported not to be present in OLs. Antisera raised against a T-cadherin peptide indicated the protein colocalized with the OL lineage markers A(2)B(5), A007, and 01 in mouse primary glial cultures. However, small round cells resembling OL precursors labeled intensely with T-cadherin, but were negative for the other markers, suggesting that this gene might be expressed earlier in the lineage. In early postnatal brain, in addition to the expected neuronal tracts, the T-cadherin antibody labeled small bipolar cells, approximately 8-10 microm in diameter, in white matter tracts. These cells had the morphology of OLs or their precursors and were identified within the cerebellar white matter and the corpus callosum, regions rich in OLs. The third clone, 3g5, was homologous to the P8 clone isolated from rat pancreas. It encoded an 80-amino-acid polypeptide with a protein kinase C domain suggesting a possible role in signal transduction. Antisera to this peptide also colocalized 3g5 with cells expressing A(2)B(5), A007, and 01 in culture and in cells within white matter tracts which had the same morphology as those labeled by T-cadherin in these regions. In addition to these, beta(10) thymosin and mevalonate kinase clones were also isolated from the screen. PMID- 11872947 TI - Inhibition of alpha(1-6)-linked fucose decreases inner retinal cells and increases photoreceptors in chicken retinal reaggregates. AB - We investigated the developmental role of alpha(1-6)-linked fucose, applying Aleuria aurantia lectin to a specific retinal regeneration system. Thereby, dissociated retinal cells of chicken embryos reaggregate, proliferate, and differentiate in vitro into histotypical spheres, so-called retinospheroids. Under the influence of A. aurantia lectin, processes of proliferation, differentiation and histogenesis of retinospheroids were disturbed. Extending these in vitro studies, we here show that A. aurantia lectin treatment decreases cells of the inner half retina and their processes into inner plexiform layer areas, as revealed by quantitative enzyme histochemistry for butyryl- and acetylcholinesterase, and immunohistochemistry using antibodies to acetylcholinesterase, Pax-6, calbindin-D, and F11. Concomitantly, the number of rod and red/green photoreceptors dramatically increases, using the antibodies rho4D2 and CERN901 (both specific for rods) and CERN906 (specific for red/green cones). These findings show that glycoproteins exhibiting fucose in alpha(1-6) linkage are involved in processes determining retinal cell fate, strongly shifting the relative ratio of cells of the inner towards cells of the outer retina. PMID- 11872949 TI - Incorporation of long CDR3s into V domains: implications for the structural evolution of the antibody-combining site. AB - Available data suggest that 'primitive' antibody-combining sites often include longer than average HCDR3s. Long HCDR3 sequences have been reported in diverse vertebrates, including humans, cattle, camels and sharks. These long HCDR3 segments contain unusual sequence features such as stretches of Gly or Pro residues and multiple Cys residues. We examined how longer than average HCDR3s were accommodated in the V domains of human, murine and camel antibodies with known three-dimensional structures. The main conclusions were that (1) HCDR3s longer than 12 residues should protrude outward from the V domains; (2) descending HCDR3 polypeptides may utilize VL (including LCDR3) constituents as a platform, supporting the protruding segments; (3) intra- and inter-HCDR disulfides are frequently formed to rigidify the structure of HCDR3 or the combining site, and (4) V and C domains were possibly more similar in primordial antibodies than they are in their present day counterparts. PMID- 11872950 TI - Analysis of the sequence polymorphism within class II transactivator gene promoters. AB - The class II transactivator is a major transcriptional factor acting on the promoters of MHC class II genes. Transcription of the CIITA gene is driven by four alternative promoters, which exhibit cell-type-specific activity. The CIITA promoter III (PIII) is constitutively active in B cells, whereas promoter IV (PIV) becomes activated upon interferon-gamma activation. The aim of this study was to investigate whether these two promoters exhibit a sequence variability like the MHC class II promoters do. We isolated PIII and PIV fragments from healthy individuals and rheumatoid arthritis patients and screened them for sequence polymorphisms. Single base pair substitutions within the CIITA PIV were found in 9% of the individuals analyzed. The majority of the substitutions were located upstream of the known cis-acting elements of the promoter. PIII was non polymorphic. To evaluate the functional relevance of the detected polymorphism we cloned variable PIV upstream of the luciferase reporter gene. Such prepared constructs were transfected into monocytes, melanoma and HeLa cells, which were subsequently stimulated with interferon-gamma. The analysis of promoter activities did not reveal significant differences in all three cell types. We conclude that the level of CIITA expression does not vary within the population. Thus the differences in the level of MHC class II expression, which are observed between individuals, stem for the polymorphisms of the MHC class II promoters themselves. PMID- 11872951 TI - Increased frequency of the C3*F allele and the Leiden mutation of coagulation factor V in patients with severe coronary heart disease who survived myocardial infarction. AB - The aim of the present study was to compare the frequencies of the F allele of C3 complement component and the Leiden mutation of coagulation factor V in patients with severe coronary heart disease (CHD) who survived myocardial infarction (MI; group A), and those who had no MI in their case history (group B). We have determined the C3 allele frequencies by electrophoresis, and Leiden mutation by PCR in 338 patients with severe CHD and in 490 and 523 healthy controls, respectively. The C3*F allele frequency was significantly (p = 0.006) higher in group A (0.213) that in group B (0.132). A significant (p = 0.045) difference was found between < or = 60-year group A (0.077) and group B (0.029) patients in the frequency of Leiden mutation. These findings indicate that the C3*F allele and the Leiden mutation may be associated with an increased risk of developing myocardial infarction in CHD patients. PMID- 11872952 TI - In situ activated intestinal T cells expanded in vitro--without addition of antigen--produce IFN-gamma and IL-10 and preserve their function during growth. AB - OBJECTIVE: The balance between mucosal CD4+ T cells producing IFN-gamma or IL-10 is essential in the maintenance of intestinal homeostasis. We aimed to investigate how in situ activated T cells were expanded in vitro according to a new cell culture protocol and if it selected for continuous clonal CD4+ T cells capable of producing mainly IFN-gamma or IL-10. METHODS: T cell cultures were established from colonic biopsy specimens of 27 patients with Crohn's disease and from 10 healthy controls in a medium supplemented with IL-2 and IL-4 but without addition of exogenous antigen or mitogen. Cytokine production was measured after stimulation (IL-12, super antigen) and inhibition (cyclosporin and methylprednisolone). RESULTS: Cytokine production was increased in cultures from patients with Crohn's disease compared to controls (IFN-gamma, p = 0.005; IL-10, p = 0.003; TNF-alpha, p = 0.03). Early cultures were highly responsive to IL-12 stimulation. We established CD4+ T-cell clones escaping cellular senescence with an inflammatory or regulatory cytokine profile. DISCUSSION: The data indicate that cultures of in situ activated inflammatory and regulatory subpopulations of intestinal T lymphocytes with pathogenic importance in Crohn's disease can be established preserving their functional properties during growth. PMID- 11872953 TI - Use of human recombinant DNase I expressed in COS-7 cells as an immunogen to produce a specific anti-DNase I antibody. AB - To obtain human deoxyribonuclease I (DNase I) as an immunogen, we have developed a procedure that is more useful and effective than the conventional procedure, which uses human urine as a starting material. In the new procedure, we culture COS-7 cells transfected with expression vector carrying human DNase I cDNA, and then purify the enzyme from the culture medium. The enzyme can be easily isolated to apparent homogeneity by passage through only three chromatography columns. The rabbit antiserum that we used against the recombinant DNase I was not inferior to that used against DNase I from human urine, in terms of both its ability to discriminate DNase I phenotypes and its ability to neutralize enzyme activity. Therefore, our procedure may be useful for producing an antibody specific for human DNase I. PMID- 11872954 TI - Oligonucleotide fishing for STAT6: cross-talk between IL-4 and chemokines. AB - Signal transducer and activator of transcription 6 (STAT6) is essential for the biological activities of interleukin-4 (IL-4) and the development of allergic responses in mice. Here we report on a sensitive and specific assay for STAT6 activation in response to IL-4. We took advantage of double-stranded oligonucleotide probes containing a STAT6-binding gene-sequence from the promotor of the immunoglobulin heavy chain germline epsilon transcript to study the IL-4 induced DNA binding of STAT6. Using these probes, we show that repeated adjacent STAT6-binding sites result in enhanced STAT6-DNA binding. Moreover, the distance between the binding sites is critical for STAT-DNA binding, i.e. STAT6 binding is decreased at distances above 20 nucleotides between neighbouring binding sites. Using this assay to study cross-talk between IL-4 and chemokines, we provide evidence that MIP-1beta and MIG inhibit IL-4-induced STAT6 activation, whereas other chemokines and cytokines do not. In conclusion, our data show that oligonucleotide fishing is a supplementary tool for studying cytokine cross-talk at a genomic level. PMID- 11872955 TI - Nomenclature of the human immunoglobulin lambda (IGL) genes. AB - 'Nomenclature of the Human Immunoglobulin Lambda (IGL) Genes', the 18th report of the 'IMGT Locus in Focus' section, provides the first complete list of all the human IGL genes. The total number of human IGL genes per haploid genome is 87--96 (93--102 if the orphons are included), of which 37--43 genes are functional. IMGT/Human Genome Organization (HUGO) gene names and definitions of the human IGL genes on chromosome 22q11.2 and IGL orphons on chromosomes 8 and 22 are provided with the gene functionality and the number of alleles, according to the rules of the IMGT Scientific chart, with the accession numbers of the IMGT reference sequences and with the accession ID of the Genome Database GDB and NCBI LocusLink databases, in which all the IMGT human IGL genes have been entered. The tables are available at the IMGT Marie-Paule page of IMGT, the international ImMunoGeneTics database (http://imgt.cines.fr) created by Marie-Paule Lefranc, Universite Montpellier II, CNRS, France. PMID- 11872956 TI - Nomenclature and overview of the mouse (Mus musculus and Mus sp.) immunoglobulin kappa (IGK) genes. AB - 'Nomenclature and overview of the mouse (Mus musculus and Mus sp.) immunoglobulin kappa (IGK) Genes', the 19th report of the 'IMGT Locus in Focus' section, provides the first complete list of all the mouse (M. musculus) IGK genes. The mouse (M. musculus) locus spans 3,200 kb. The total number of mouse (M. musculus) IGK genes per haploid genome is 164 (174 if the orphons are included). The functional genomic repertoire comprises 93 IGKV belonging to 18 subgroups, 5 IGKJ and 1 IGKC gene. IMGT gene names and definitions of the mouse (M. musculus) IGK genes on chromosome 6 and IGK orphons are provided with the gene functionality and the number of alleles, according to the concepts of IMGT-ONTOLOGY and to rules of the IMGT Scientific chart, with the accession numbers of the IMGT reference sequences. These tables and figures are available at the IMGT Marie Paule page of IMGT, the international ImMunoGeneTics database (http://imgt.cines.fr) created by Marie-Paule Lefranc, Universite Montpellier II, CNRS, France. PMID- 11872957 TI - Autophagy and nuclear changes in FM3A breast tumor cells after epirubicin, medroxyprogesterone and tamoxifen treatment in vitro. AB - OBJECTIVE: Autophagy is a form of physiological programmed cell death which is observable after hormonal withdrawal. In this study, the FM3A murine breast tumor cell line was treated with epirubicin alone and with medroxyprogesterone acetate (MPA) or tamoxifen, to determine if structural and kinetic signs of autophagy may also occur in an enhanced manner during epirubicin sensitization via hormonal agents. METHODS: One-week soft agar colony growth, 96-hour values of plating and pulse thymidine labeling and electron microscopical examinations were performed following treatment with MPA and tamoxifen alone or with epirubicin. RESULTS: Tamoxifen induced signs of autophagy, which was enhanced when it was combined with MPA. Epirubicin also induced autophagy of secretory granules, which coalesced to form an intracytoplasmic lumen. Combining MPA with epirubicin enhanced the formation of apoptotic blebs and chromatin fragmentation. When epirubicin was combined with tamoxifen, peculiar nuclear structures were formed. CONCLUSIONS: Hormonal agents may modulate anthracycline activity towards specific patterns in eliciting cell death, via autophagy and/or as yet unknown nucleolus specific interactions. PMID- 11872958 TI - Overexpression of sialyl Lewis x antigen is associated with formation of extratumoral venous invasion and predicts postoperative development of massive hepatic metastasis in cases with pancreatic ductal adenocarcinoma. AB - The clinicopathological factors and expression of sialyl Lewis antigens which are the cell adhesion molecules to endothelial cells were compared in relation to the extent of the postoperative hepatic metastasis in 23 consecutive patients with pancreatic ductal adenocarcinoma whose clinical courses were carefully monitored and documented. The overall survival of cases with massive hepatic metastasis (MHM) was significantly poorer than that of those with local or no hepatic metastasis (p = 0.0453). Postoperative MHM was significantly correlated with the presence of duodenal invasion (p = 0.0418), the presence of portal vein invasion (p = 0.0435), the presence of extratumoral venous invasion (p = 0.0052) and high expression of sialyl Lewis x antigen (p = 0.0022). Multivariate analysis confined significant correlation between the high expression of sialyl Lewis x antigen and the development of MHM (p = 0.0402). Kaplan-Meier analysis revealed that the overall survival of patients with a high expression of sialyl Lewis x antigen was significantly poorer than that of patients with a low expression of the antigen (p = 0.0216). These results indicate that the overexpression of sialyl Lewis x antigen plays an important role in the development of MHM, and also predicts a poorer overall survival of these patients. Further studies with more cases are warranted to confirm these results. PMID- 11872959 TI - PTEN expression in breast and endometrial cancer: correlations with steroid hormone receptor status. AB - OBJECTIVE: The PTEN (MMAC1/TEP1) tumor suppressor gene is frequently mutated and homozygously deleted in human neoplasms, but there is only sparse information about PTEN protein expression in hormone-dependent female tumors. Therefore, we investigated PTEN expression in 68 breast and 43 endometrial carcinomas. METHODS: For PTEN protein detection, we used Western blot analysis followed by densitometry and compared these data with clinicopathologic parameters, the estrogen receptor (ER) and progesterone receptor (PR) status, HER2/neu and the proliferation marker Ki67. RESULTS: We were able to show significantly decreased PTEN protein expression in endometrial carcinomas compared with normal endometrial tissue samples, especially in the endometrioid histological subtype. In contrast, PTEN downregulation was found more rarely in breast cancer. Lower PTEN expression in breast cancer correlated significantly with high ER immunoreactivity (p = 0.008) and was weakly associated with PR expression (p = 0.055) and low histological grading (p = 0.081). No correlation with any of these parameters was observed in endometrial tumors. In both tumor types, no association of PTEN expression with any other analyzed parameter was found. CONCLUSIONS: These results suggest that PTEN expression plays different roles in the pathogenesis of endometrial carcinomas and breast cancer. In mammary carcinomas, loss of PTEN expression is mainly found in more differentiated tumors and is probably not a major event in carcinogenesis. PMID- 11872960 TI - Promoter methylation status of the DNA repair genes hMLH1 and MGMT in gastric carcinoma and metaplastic mucosa. AB - Hypermethylation of CpG islands in the promoter region is associated with the silencing of a variety of tumor suppressor genes. DNA repair genes human Mut L homologue 1 (hMLH1) and O(6)-methylguanine-DNA methyltransferase (MGMT) have been shown to be hypermethylated in certain carcinomas. We studied DNA methylation of CpG islands in hMLH1 and MGMT in 50 gastric carcinomas and 10 intestinal metaplastic mucosa samples. We analyzed the methylation status of hMLH1 and MGMT using methylation-specific polymerase chain reaction and DNA sequencing analysis. We measured protein levels of hMLH1 using Western blot and immunohistochemical analysis. CpG island hypermethylation of hMLH1 and MGMT was detected in 11 (22%) and 8 (16%) of the 50 gastric tumors, respectively. Hypermethylation of the promoter was more common in intestinal-type gastric carcinomas than in poorly diffuse-type gastric carcinomas (p = 0.016 and 0.021, respectively; Fisher's exact test). However, hMLH1 promoter hypermethylation did not coincide with MGMT promoter hypermethylation except in 1 patient. Hypermethylation of the hMLH1 promoter but not the MGMT promoter occurred in intestinal metaplastic mucosae. Immunohistochemical analysis revealed a corresponding reduction in hMLH1 protein expression in some of the intestinal metaplastic mucosae. Our results suggest that at least two types of promoter methylation participate in the development of gastric carcinoma. Tumor-specific promoter hypermethylation of hMLH1 may be an early event in carcinogenesis in the stomach. PMID- 11872961 TI - Expression of minichromosome maintenance 2 (MCM2), Ki-67, and cell-cycle-related molecules, and apoptosis in the normal-dysplasia-carcinoma sequence of the oral mucosa. AB - OBJECTIVE: We examined cell cycle and cell death biomarker trends with the normal dysplasia-carcinoma sequence of the oral epithelia analyzing the pathological significance of a new biomarker, minichromosome maintenance 2 (MCM2). METHODS: This study analyzed 12 patients with normal oral epithelia, 69 with dysplasia, and 35 with squamous cell carcinoma (SCC); in 13 patients, SCCs were preceded by dysplasia. The sections were immunostained for MCM2, Ki-67, P53, P27(Kip1) and P21(CIP1/WAF1), and conducted by TUNEL methods. Western blot analysis of MCM2 was performed in the 4 human cultured oral SCCs, all of which showed the expression. RESULTS: Significantly higher labeling indices (LI; %) of MCM2, Ki-67, and P53, as well as lower LI of TUNEL indices (TI; %), P27, and P21 were noted in the SCCs than in the dysplasias. The 13 dysplasias developed SCC with significantly higher LI of MCM2 and P53, and lower LI of P21 than the other dysplasias (each p < 0.05). The LI of MCM2, P21 and the TI were not correlated with P53 expression. CONCLUSIONS: Oral dysplasia was characterized by lower cell proliferation and a higher frequency of cell death compared to SCCs. The higher LI of MCM2 and P53 and the lower LI of P21 might predict malignant transformation of oral dysplasia. MCM2 is regulated via a P53-independent pathway, and a useful biomarker of proliferating cells. PMID- 11872962 TI - Differential adhesion of polymorphous neutrophilic granulocytes to macro- and microvascular endothelial cells under flow conditions. AB - OBJECTIVE: As one of the important active barriers in the human organism, endothelial cells (EC) play a central role in the biological reaction to a variety of stimuli, e.g. during the induction and regulation of inflammation, as well as in the reaction to transplantation and biomaterial implantation. In the study of endothelial function, the most widely used in vitro model is that of human umbilical vein EC (HUVEC), i.e. an EC type of embryonic and macrovascular origin. However, many of the important pathological processes occur at microvascular level, thus questioning the validity of the HUVEC model. Moreover, the morphological and functional heterogeneity of the endothelium in the various organs, e.g. kidney, liver and lung, must be taken into consideration. The purpose of the present study was to use a dynamic cell culture system to compare the reactions of HUVEC and human pulmonary microvascular EC (HPMEC) to pro inflammatory stimulation. METHODS: HUVEC and HPMEC in monolayer culture were stimulated by tumor necrosis factor-alpha (TNFalpha) in a parallel-plate flow chamber. Short- (4 h) and long-term (12 h) stimulation were compared. As a functional parameter, the adhesion of human peripheral blood polymorphonuclear granulocytes (PMN) to EC was quantitated both under venous and arterial flow conditions. RESULTS: Short-term (4 h) TNFalpha stimulation and venous flow conditions elicited a 32% higher PMN adhesion to HPMEC compared with HUVEC, whereas under arterial flow conditions no statistically significant differences were found. Following longer-term (12 h) TNFalpha stimulation, PMN adhesion to HPMEC was 65% higher than to HUVEC under venous flow. Under arterial flow no differences were detected. CONCLUSION: The present results provide new data on the heterogeneity of the endothelium and affect a central element in microvascular pathology, namely granulocyte-endothelial interactions. Moreover, this paper emphasizes the necessity to evaluate the in vitro models of the endothelium with respect to the extrapolation to the situation in vivo. PMID- 11872963 TI - Decreased expression of E-cadherin and Yamamoto-Kohama's mode of invasion highly correlates with lymph node metastasis in esophageal squamous cell carcinoma. AB - OBJECTIVES: A reduction in cell-cell adhesion in cancer cells is an essential step in the progression from localized malignancy to metastatic disease. E Cadherin is an important component of cell-cell adhesion molecules and may be a crucial determinant of tumor invasion and metastasis. E-Cadherin expression is reported to be correlated with lymph node metastasis in esophageal squamous cell carcinoma (SCC). The objective of this experiment is to examine the factors that are associated with invasion and metastasis of esophageal SCC. METHODS: Forty-six cases of esophageal SCC were examined by immunohistochemical staining for E cadherin. The relationship between E-cadherin-staining patterns, conventional clinicopathological parameters and Yamamoto-Kohama's (Y-K's) mode of invasion were examined. RESULTS: The expression of E-cadherin on the cell membrane was reduced or lost in some of the esophageal SCC. Lymph node metastasis was highly correlated with the expression pattern of E-cadherin (p = 0.0002) and also highly correlated with Y-K's mode of invasion (p = 0.0078). However, lymph node metastasis was not correlated with any conventional clinicopathological parameters for invasion. CONCLUSIONS: These results indicate that E-cadherin plays a crucial role in invasion and metastasis in esophageal SCC, and that Y-K's mode of invasion highly reflects the invasiveness and metastatic potentials of esophageal SCC cells. Therefore, examination of the expression of E-cadherin and Y-K's mode of invasion would be helpful in predicting lymph node metastasis in esophageal SCC. PMID- 11872964 TI - The changing pattern of coronary heart disease in the elderly. PMID- 11872965 TI - Prospects for prevention of coronary disease in the elderly. Introduction. PMID- 11872966 TI - Epidemiologic appraisal of hypertension as a coronary risk factor in the elderly. AB - Five decades of epidemiologic research has established elevated blood pressure as a major contributor to atherosclerotic cardiovascular diseases in the elderly, including coronary heart disease. Clinicians formerly favored the diagnosis and treatment of hypertension in terms of the diastolic blood pressure and categorical "hypertension." Epidemiologic data now emphasize the essential role of systolic blood pressure, pulse pressure, and a graded influence of blood pressure, even within the high-normal range. The risk of coronary heart disease, the most common lethal sequela of hypertension, increases with the extent of risk factor clustering. Among hypertensive persons, about 39% of coronary events in men and 68% in women are attributable to the presence of two or more additional risk factors. When risk factor clustering is associated with glucose intolerance, obesity, and dyslipidemia, it may be attributed to insulin resistance promoted by abdominal obesity. Other hazardous influences often accompanying hypertension in the elderly are the presence of an elevated heart rate, elevated levels of fibrinogen, and left ventricular hypertrophy. Because clustering with other risk factors is characteristic of hypertension in the elderly, it is essential to screen for them and for the presence of comorbid cardiovascular diseases, target organ disease, and subclinical vascular disease likely to be present. Multivariate risk assessment profiles enable global estimation of hypertensive risk of developing coronary heart disease. Hypertensive elderly patients are more appropriately targeted for antihypertensive therapy by such risk stratification than by relying solely on the severity of the blood pressure elevation. The goal of therapy should be to improve the multivariate risk profile as well as the level of the blood pressure. PMID- 11872968 TI - Coronary heart disease risk factors in the elderly. AB - Hypertension, dyslipidemia, impaired glucose tolerance, and obesity remain the major modifiable risk factors for most of the coronary disease afflicting the elderly. The relative risk associated with these established risk factors diminishes with advancing age, but this is offset by a greater absolute and attributable risk. Diabetes is increasing alarmingly in prevalence and operates more powerfully in women, eliminating their coronary disease resistance (relative to men). Interest in this entity now focuses on the insulin resistance syndrome promoted by abdominal obesity that has become so common in the elderly. The isolated systolic hypertension and large pulse pressure that predominate in the elderly is now recognized as a coronary disease hazard. Dyslipidemia, characterized by a high total to high-density lipoprotein cholesterol ratio, is the most predictive lipid profile for coronary disease in the elderly. High triglycerides, accompanied by low high-density lipoprotein cholesterol usually signifies insulin resistance and more atherogenic, small, dense low-density lipoprotein. Left ventricular hypertrophy is an ominous harbinger of coronary disease. Fibrinogen and the leukocyte count are correlated coronary disease risk factors that may indicate unstable lesions. Novel risk factors, such as hemostatic factors, homocysteine, lipoprotein(a), C-reactive protein, and hyperinsulinemia, are worthy of attention, but the efficacy of correcting them in the elderly has not yet been demonstrated. Nor has the efficacy of hormone replacement therapy in women. All the coronary risk factors tend to cluster, and the hazard posed by each is greatly influenced by the burden of coexisting risk factors. High-risk elderly candidates for coronary disease can be efficiently targeted for treatment by global risk assessment, using only the major established risk factors. The distinction between primary and secondary prevention in the elderly is less clear than in the middle-aged because they often have advanced presymptomatic vascular pathology that imposes a coronary event rate comparable to that of the middle-aged who have already sustained a clinical event. Declines in coronary mortality rates in the United States have included the elderly, justifying optimism about the efficacy of preventive measures. Most of the elderly have sufficient remaining life expectancy to warrant vigorous preventive management. Trials of risk factor modification in the elderly indicate that decades of exposure to modifiable risk factors can be countered by measures implemented late in life. PMID- 11872967 TI - Hemostatic and inflammatory markers as risk factors for coronary disease in the elderly. AB - Inflammation has been recognized as an integral component of atherothrombotic disease, and inflammatory markers are strongly related to future cardiovascular disease risk in elderly men, and to a certain extent in elderly women. The association is complicated by the prevalence of hormone replacement therapy among older women, and the variety of the underlying vascular disease. For fatal events, inflammation markers exhibit a time-to-event dependency in the elderly that has not been noted to the same degree in younger people. Hemostatic markers that represent inflammatory processes or coagulation activation also predict future risk in older individuals, but there is little evidence for associations between cardiovascular disease risk and ambient levels of coagulation or fibrinolytic factors. PMID- 11872969 TI - Current practice and future promise for clinical noninvasive measurements of subclinical atherosclerotic disease in the elderly. AB - The detection, treatment, and follow-up of subclinical vascular disease are becoming clinically essential components of cardiovascular disease prevention in the elderly. Noninvasive measurements are available for different vascular beds, including carotid, coronary, aortic, and peripheral arterial circulation. Current interest in these measures is aimed at improving the accuracy of risk prediction for coronary heart disease and cardiovascular disease. Indirect physical examination and imaging evaluations detect significant obstruction of flow in the peripheral arteries. Doppler measures of ankle-arm blood pressure index represent a simple, indirect test that has been shown to be predictive of incident cardiovascular disease independent of risk factors. Newer, high-resolution tests allow direct detection and quantitation of the burden of atherosclerosis and vascular disease within the arterial wall, independent of flow obstruction. Carotid intimal-medial thickness predicts incident coronary heart disease and stroke in the elderly, even after adjustment for traditional risk factors. Coronary calcium can be accurately detected by computed tomography and is a strong predictor of the incidence of coronary heart disease events. Evidence is accruing that coronary calcium screening will play a role in prevention in the elderly. Magnetic resonance imaging is currently under study as a promising modality for detection and quantification of aortic and carotid plaque. Ongoing studies will provide important information regarding the appropriate role of the many newer, high-resolution tests of subclinical atherosclerosis in disease prediction, treatment, and tracking of disease progression in the elderly. PMID- 11872970 TI - Obesity, diabetes, and risk of cardiovascular disease in the elderly. AB - There is an age-related increase in total body fat and visceral adiposity until age 65 years that often is accompanied by diabetes or impaired glucose tolerance. The prevalence of type 2 diabetes increases progressively with age, peaking at 16.5% in men and 12.8% in women at age 75-84 years. Over age 65, diabetes or glucose intolerance was present in 30%-40% of Framingham Study subjects. There has been an alarming increase, of epidemic proportions, in both obesity and diabetes in the general population. Type 2 diabetes and obesity are both associated with a clustering of atherogenic risk factors, and when three or more are present it generally signifies an insulin resistance syndrome. This is promoted by weight gain and visceral adiposity. The risk of macrovascular disease is increased before glucose levels reach the diagnostic threshold for "diabetes," and 25% of newly diagnosed diabetics already have overt cardiovascular disease. In the Framingham Study, increased risk of cardiovascular disease was two-fold in men and three-fold in women, eliminating the female advantage over men for all outcomes except stroke. Coronary disease is the most common and lethal sequela, and unrecognized myocardial infarctions are three times more common in diabetic than nondiabetic men. Following a myocardial infarction, diabetes imposes a high rate of recurrence, heart failure, and death, more so in women than men. The risk of cardiovascular sequelae in diabetics is variable, the majority of events occurring in those with two or more additional risk factors. Because of the variable risk of cardiovascular disease in either the diabetic or obese person, risk stratification is necessary to determine the hazard of impending cardiovascular disease. This is readily accomplished with Framingham cardiovascular risk formulations. For persons with diabetes or obesity, the chief goal is to avoid the common cardiovascular sequelae. Comprehensive care should include not only normalization of the blood sugar, but also weight reduction, dietary fat restriction, strict blood pressure and lipid control, exercise, and avoidance of tobacco. Trial data indicate that preventive measures benefit obese diabetics even more than nondiabetics. PMID- 11872972 TI - Atrial tachycardia with AV nodal Wenckebach in digitalis excess. PMID- 11872971 TI - Ethical issues in the management of geriatric cardiac patients: a competent patient on renal dialysis asks to go home to be under the care of her unqualified husband. PMID- 11872973 TI - Massive calcific deposits in the epicardial coronary arteries in the absence of calcific deposits in the aortic valve cusps and in the mitral valve annulus. PMID- 11872974 TI - Left atrial thrombus in dilated cardiomyopathy. PMID- 11872976 TI - Barriers to participation in and adherence to cardiac rehabilitation programs: a critical literature review. AB - Despite the documented evidence of the benefits of cardiac rehabilitation (CR) in enhancing recovery and reducing mortality following a myocardial infarction, only about one third of patients participate in such programs. Adherence to these programs is an even bigger problem, with only about one third maintaining attendance in these programs after 6 months. This review summarizes research that has investigated barriers to participation and adherence to CR programs. Some consistent factors found to be associated with participation in CR programs include lack of referral by physicians, associated illness, specific cardiac diagnoses, reimbursement, self-efficacy, perceived benefits of CR, distance and transportation, self-concept, self-motivation, family composition, social support, self-esteem, and occupation. Factors associated with non-adherence include being older, female gender, having fewer years of formal education, perceiving the benefits of CR, having angina, and being less physically active during leisure time. However, many of the studies have methodologic flaws, with very few controlled, randomized studies, making the findings tentative. Problems in objectively measuring adherence to unstructured, non-hospital-based programs, which are an increasingly popular alternative to traditional programs, are discussed. Suggestions for reducing barriers to participation and adherence to CR programs, as well as for future research aimed at clearly identifying these barriers, are discussed. PMID- 11872977 TI - The role of the nurse in enhancing quality of life in patients with an implantable cardioverter-defibrillator: the Swedish experience. AB - During the last 10-15 years, the implantable cardioverter-defibrillator (ICD) has become an important mode of treatment for patients suffering from grave ventricular arrhythmias, but ICD implantation involves psychosocial adjustments for both patients and relatives. The aim of this pilot study was to design a plan of education and to follow a selected group of patients with interviews, observations, and a questionnaire. The goals included seeing how well they accepted their situation after the operation when they had ongoing support of the nurse, in comparison to a control group who received conventional patient education by the physician. The patients were randomly allocated into two groups. Twenty patients were recruited, 10 in the study group and 10 in the control group, between February, 1997 and April, 1998. There were 16 men (average age, 63) and four women (average age, 57). The Nottingham Health Profile was used to measure health-related quality of life. Sleep disturbances were the greatest problem in both the study group and the control group before ICD implantation. In the study group, there was a significant improvement (p<0.05) after ICD implantation in four patients. The study also revealed a difference between men and women, with women having more sleep disturbances before ICD implantation than men (p<0.05). In both groups, there was a lack of energy and emotional reactions, both before and after ICD implantation. Few considered family life a problem before or after the study. In the control group, the patients missed the lack of contact with health care personnel more than in the study group. There was also a greater need for group meetings after the hospital stay. By means of the questionnaire, interviews, and observations, it became evident that there was a great need for information, and a plan of patient education in addition to follow up by the nurse was felt to be very important. PMID- 11872978 TI - Current options in the diagnosis and management of acute limb ischemia. AB - Acute limb ischemia occurs due to a sudden decrease in the blood flow to a limb, resulting in a potential threat to the viability of the extremity. Unfortunately, the threat is not only to the limb, but these patients are also at high risk for death. Limb hypoperfusion results in systemic acid-base and electrolyte abnormalities that impair cardiopulmonary and renal function. Successful reperfusion may result in the release of highly toxic free radicals, further compromising these critically ill patients. Therapeutic choices are often few and patient expectations are not always realistic. The management of acute limb ischemia requires a thorough understanding of the anatomy of the arterial occlusion and the open surgical and percutaneous options for restoring limb perfusion. Priorities for the diagnosis and effective management of these critically ill patients are provided. PMID- 11872979 TI - Emergent cardiovascular risk factor: homocysteine. AB - Homocysteine is an independent, modifiable risk factor for cardiovascular disease. It is an intermediate amino acid formed during the metabolism of methionine. Plasma homocysteine is normally < or = 12 micromol/L, but when elevated has many deleterious cardiovascular effects. This review explains homocysteine metabolism, the effects of elevated homocysteine, factors contributing to high homocysteine, and its measurement. Risk factors for elevated homocysteine and intervention with B vitamins are discussed. Cardiovascular nurses are encouraged to facilitate homocysteine awareness through a variety of educational means. PMID- 11872980 TI - African Americans' knowledge of and attitudes toward coronary artery bypass surgery as a treatment option. AB - The objective of this study was to determine if African Americans possess knowledge of cardiac bypass surgery as a treatment option, and their willingness to undergo the procedure if it were recommended by a cardiologist. Primary data were collected over a 2-week period. The study setting was a primary care clinic located in an urban area. The clinic is associated with a university medical center that provides high-technology cardiac services. An exploratory, prospective survey and interview technique was utilized to elicit responses from a convenience sample of 50 African American participants. The majority (80%) were knowledgeable about coronary artery bypass surgery and stated a positive preference for the procedure, if it were recommended. The author concluded that although African Americans are knowledgeable about coronary bypass, awareness of these procedures as a treatment option needs to be supported by nurses and physicians. PMID- 11872981 TI - Nurses' initiatives in smoking cessation in Hong Kong. PMID- 11872982 TI - How may we evaluate and treat a wide-complex tachycardia in a patient presenting with chest pain in the emergency department? PMID- 11872986 TI - Reducing the knowledge-practice gap in the management of patients with cardiovascular disease. AB - Despite major progress in the development of effective therapy to reduce mortality and morbidity from cardiovascular disease, it remains the leading cause of mortality in this country. One aspect of this problem is represented by the lag in adoption of treatments with documented efficacy in large clinical trials. This "knowledge-practice gap" has been attributed to factors at multiple levels of the health care system that impede implementation of optimal therapy. Although there is evidence of progress in the use of recommended therapeutic modalities in the past decade, this has been modest. Recent approaches to assessment of patient care by physicians, health plans, and institutions through the tracking of clinical performance have been instituted to promote optimal patient care. They are being increasingly utilized for purposes of accreditation and will also provide guidance for consumer purchasing of health care. Such methods have the potential to promote increased adherence to current standards of care. PMID- 11872987 TI - The chest pain center strategy for delivering community heart attack care by shifting the paradigm of heart attack care to earlier detection and treatment. AB - Heart attack remains the number one health problem in the United States and throughout the world. It has been that way for more than 100 years. Unless we change our course, heart attack will continue to exert its horrendous casualties, not only in the United States but also throughout the world. Our present strategy in dealing with this problem needs both leadership and a change in direction. In an effort to search "outside the box" for the solution to this problem, this symposium is a call to action that challenges us to approach the heart attack problem with a mindset bent on winning this war against heart disease, and not coexisting and accepting the problem as an inescapable fate. PMID- 11872988 TI - A comparison trial for stratifying intermediate-risk chest pain: benefits of emergency department observation centers. AB - Chest pain of uncertain etiology (intermediate-risk chest pain [IR-CP]) constitutes a majority of acute chest pain presentations to emergency departments (EDs). A before- and-after trial of 2197 IR-CP patients transferred from the hospital's ED to one of three units-ED-based observation center (ED-OC), inpatient observation center (IN-OC), and inpatient units-compared mean cost, length of stay, and safety over a 2-year period. The mean per patient cost for management of IR-CP was lower in the ED-OC ($1642) than the IN-OC ($1910) or the inpatient units ($2785). The mean length of stay was shorter in the ED-OC (0.75 days) than in the IN-OC (1.18 days) or the inpatient units (2.16 days). Return rates were lower in the ED-OC at 7 days (0%) and at 6 months (0.45%) than the IN OC (0% and 1.22%) or the inpatient units (0.77% and 3.67%). Overall hospital costs for managing IR-CP dropped significantly (12.5%) after the ED-OC was opened. ED-OCs provide a safe and cost-effective alternative to admission of IR CP patients. PMID- 11872983 TI - Detection of subclinical atherosclerosis using noninvasive tools. PMID- 11872989 TI - Conquering heart disease: a call to action. AB - Despite 20th century scientific and technologic advances in the area of cardiovascular medicine and public health and associated benefits in terms of significant declines in mortality, heart disease remains as the dominant cause of death for all Americans. Tremendous geographic variation in age-adjusted coronary heart disease mortality, varying over two-fold between the highest and lowest states, provides troubling evidence that this epidemic is raging unrelentingly in many communities across our country. Also disturbing are signs that the significant decline in coronary heart disease mortality observed since the 1960s may have slowed in recent years. The coronary heart disease epidemic continues to hit hard among certain racial and ethnic minorities, prompting the Healthy People 2010 national call for action to eliminate cardiovascular disease disparity. PMID- 11872990 TI - Dialing 911: a call to action. AB - Cardiovascular disease and stroke are the nation's leading causes of disability and death. Scientific advances have provided newer treatments that, when applied effectively and in time, save a significant number of lives. Multiple factors in our system of delivery impede our ability to apply effective treatments, but with an analysis of the components of care, each of us can take leadership roles in our communities to improve the chain of survival. These elements and the science supporting each are briefly reviewed. PMID- 11872991 TI - C-reactive protein, statins, and the primary prevention of atherosclerotic cardiovascular disease. AB - Emerging data implicate inflammation as integral to atherosclerosis and its complications. From a clinical perspective, the inflammatory biomarker C-reactive protein has demonstrated consistent predictive value in the detection of individuals at high risk for cardiovascular disease. Therapy with 3-hydroxy-3 methylglutaryl coenzyme A reductase inhibitors (statins) reduces C-reactive protein as well as low-density lipoprotein cholesterol, thus providing a potential additional mechanism for the reduction in cardiovascular events associated with the use of these agents. Evidence from the Air Force/Texas Coronary Atherosclerosis Prevention Study suggests that statin therapy may be effective in reducing incident coronary events among those with elevated levels of C-reactive protein but normal levels of low-density lipoprotein cholesterol. These data, along with accumulating laboratory data, support a potential anti inflammatory benefit of statins. Large-scale, randomized trials in the primary prevention of acute coronary events among individuals without overt hyperlipidemia but with evidence of elevated C-reactive protein are now needed to directly test this hypothesis. PMID- 11872992 TI - Still more benefits of statins: initial observations in aortic stenosis. PMID- 11872993 TI - Analysis of the mitochondrial DNA genome in the peripheral blood leukocytes of HIV-infected patients with or without lipoatrophy. AB - OBJECTIVE: To investigate the molecular mechanisms of nucleoside analogue reverse transcriptase inhibitor (NRTI)-associated mitochondrial dysfunction. METHODS: Peripheral blood samples were collected from 10 healthy individuals, 10 HIV infected, NRTI-treated patients with lipoatrophy, and four HIV-infected patients naive to all antiretrovirals. DNA was isolated from the leukocytes and the mitochondrial genome analyzed for DNA depletion, deletions and point mutations. RESULTS: We were not able to detect mitochodrial DNA (mtDNA) depletion, deletions, or DNA rearrangements in any of the specimens, including one from a patient with fulminant lactic acidosis. A complete analysis of the entire mitochondrial genome by temporal temperature gradient gel electrophoresis revealed several nucleotide substitutions in blood mtDNA of several HIV infected patients. CONCLUSION: We found no evidence for NRTI-associated mtDNA depletion or gross mtDNA mutations in leukocytes of HIV-infected patients, regardless of their treatment history. Thus, either NRTI-induced mutations in mtDNA are tissue specific or alternatively, pre-existent mtDNA variations in HIV disease predispose to the development of clinically apparent mitochondrial dysfunction during NRTI therapy. The significance of mtDNA variations in the development of mitochondrial-related clinical conditions in HIV patients with or without NRTI therapy is to be further investigated. PMID- 11872994 TI - CTLA-4 upregulation during HIV infection: association with anergy and possible target for therapeutic intervention. AB - OBJECTIVE: To study the role of cytotoxic T-lymphocyte-associated antigen 4 (CTLA 4) during HIV infection. METHODS: Intracellular CTLA-4 expression, determined by flow-cytometry, and proliferative responses to HIV antigens, were studied in peripheral blood mononuclear cells (PBMC) from 93 HIV-1-infected [HIV(+)] patients and 40 HIV-1 seronegative controls. RESULTS: The proportions of CTLA-4 expressing CD4+ T cells were: (1) significantly higher in HIV(+) patients, 10.95 +/- 0.66%, than in controls, 6 +/- 0.45% (P < 0.0001); (2) inversely correlated to CD4+ counts (r = -0.67, P < 0.005, n = 16, drug-naive patients; r = -0.57, P < 0.0001, n = 77, HAART-treated patients); and (3) positively correlated to proportion of activated (HLA-DR+CD3+) (r = 0.53, P < 0.0001) and memory (CD45RO+CD4+) T cells (r = 0.46, P < 0.001). CD28 median fluorescence intensity in CTLA-4- cells was twice that in CTLA-4+ cells (140 +/- 5.3 versus 70 +/- 2.28, P < 0.00001), whereas cells low in CD28 and CD4, expressed more CTLA-4 (P < 0.0001). Higher proportion of CTLA-4+CD4+ cells expressed CCR5 and Ki-67, in comparison with CTLA-4-CD4+ cells, (65 +/- 11.9 and 25 +/- 7.5% versus 27 +/- 8.9 and 3.7 +/- 2%, P < 0.0001 and P < 0.01, respectively). Among HAART-treated patients, with viral load below detectable levels, CD4+ cells increase was inversely correlated to %CTLA-4+CD4+ cells (r = -0.5, P = 0.003, n = 39). Proliferation of PBMC to anti-CD3, gp-120 depleted HIV-1 antigen or HIV-1 p24 stimulation was inversely correlated with CTLA-4 levels (r = -0.68, P = 0.0035; r = -0.38,P = 0.04; and r = -0.43, P = 0.028, respectively). CONCLUSIONS: (1) CTLA 4 is upregulated during HIV infection and may therefore account for CD4 T-cell decline and anergy in HIV-1 infection. (2) Increased levels of CTLA-4 may undermine immune responses and in the HAART-treated patient-immune reconstitution. (3) Blocking of CTLA-4 may offer a novel approach for immune based therapy in HIV infection. PMID- 11872995 TI - Effects of combination chemotherapy and highly active antiretroviral therapy on immune parameters in HIV-1 associated lymphoma. AB - OBJECTIVE: To measure the effects of combined chemotherapy and highly active antiretroviral therapy (HAART) on immune cell counts and plasma HIV-1 RNA loads in patients with AIDS-related lymphoma (ARL) to determine the implications for opportunistic infection prophylaxis and medium-term immune function. DESIGN AND METHODS: Peripheral blood total lymphocyte count, CD4 T-cell count, CD8 T-cell count, CD19 B-cell count, CD16/CD56 natural killer cell count and plasma HIV-1 RNA load were prospectively measured at ARL diagnosis, at 1 and 3 months during and 1, 3 and 6 months after chemotherapy in twenty patients receiving HAART. RESULTS: Significant declines in T-helper cell (CD4) count, natural killer cell (CD16/CD56) and B lymphocyte count (CD19 cells) occurred during the first 3 months of chemotherapy. There was no significant alteration in the T-cytotoxic cell (CD8) count, CD4 percentage or HIV-1 RNA load during the study period. The T helper cell and natural killer cell counts recovered to pre-treatment levels within 1 month of finishing chemotherapy. The recovery of B-cells was slower with pre-treatment levels only being achieved after 3 months. The recovery of CD4 T cell count following completion of chemotherapy was more rapid than described for ARL patients who were not receiving concomitant HAART. CONCLUSIONS: By combining chemotherapy with HAART, immune function is better maintained in the medium term. The CD4 T-cell count falls by 50% during chemotherapy and this will help to identify patients who require opportunistic infection prophylaxis during chemotherapy. PMID- 11872996 TI - Genotypic analysis of plasma HIV-1 RNA after influenza vaccination of patients with previously undetectable viral loads. AB - OBJECTIVE: In this study we evaluated the possibility that plasma viral load elevations secondary to influenza vaccination in HIV-1-seropositive individuals with previously undetectable viral loads (< 200 copies/ml) could develop resistance-bearing mutations in the viral reverse transcriptase (RT) and protease regions. METHODS: Thirty-four patients with undetectable viral burdens on highly active antiretroviral therapy (HAART) were evaluated for elevations in plasma viral load 2 and 4 weeks post-influenza vaccination. Plasma from patients whose viral load increased after vaccination was subject to genotypic resistance analysis by the line probe assay (LiPA) to determine whether primary resistance bearing mutations developed during this period and at follow-up. Stored plasma was used to evaluate whether RT or protease mutations existed pre-vaccination. RESULTS: Seven out of 34 patients were found to experience elevations in their viral load after influenza vaccination. Two of the patients revealed evidence of primary RT or protease mutations not demonstrated in earlier pre-vaccination samples. One patient failed therapy after vaccination, and one patient revealed post-vaccination viral load elevations that eventually led to the progressive development of primary zidovudine mutations. CONCLUSION: Evidence is presented that supports the contention that a small subset of patients who experience viral load elevations after influenza vaccination can develop mutational changes in the RT region of the viral genome either acutely or after a failure of the viral load to return to undetectable levels. PMID- 11872997 TI - The effects of cannabinoids on the pharmacokinetics of indinavir and nelfinavir. AB - BACKGROUND AND OBJECTIVES: The use of cannabinoids for appetite stimulation and the management of wasting and antiretroviral side-effects has become a common practice in the care of HIV-infected individuals. We present pharmacokinetic data from a randomized placebo-controlled study designed to evaluate the metabolic effects of smoked marijuana and dronabinol in HIV-infected patients receiving indinavir (IDV) or nelfinavir (NFV). METHODS: Subjects on stable regimens containing IDV 800 mg every 8 h (n = 28) or NFV 750 mg three time a day (n = 34) were randomized to one of three treatment arms: 3.95% THC marijuana cigarettes, dronabinol 2.5 mg capsules or placebo capsules administered three times daily. Serial blood sampling was performed at baseline and on day 14 of treatment. The changes in NFV and IDV pharmacokinetics were measured as the median percentage change from baseline. RESULTS: At day 14, the 8-h area under the curve (AUC(8)) changed by -10.2% (P = 0.15), maximum concentration (C(max)) by -17.4% (P = 0.46), and minimum concentration (C(min)) by -12.2% (P = 0.28) for patients in the NFV marijuana arm (n = 11). Similar decreases had occurred by day 14 among patients in the IDV marijuana arm (n = 9): AUC8 had changed by -14.5% (P = 0.074), C(max) by -14.1% (P = 0.039), and C(min) by -33.7% (P = 0.65). CONCLUSION: Despite a statistically significant decrease in C(max) of IDV in the marijuana arm, the magnitude of changes in IDV and NFV pharmacokinetic parameters in the marijuana arm are likely to have no short-term clinical consequence. The use of marijuana or dronabinol is unlikely to impact antiretroviral efficacy. PMID- 11872998 TI - Concentration-controlled compared with conventional antiretroviral therapy for HIV infection. AB - OBJECTIVES: To demonstrate the feasibility of a concentration-controlled approach to combination antiretroviral therapy, and to compare the virological responses and safety of this strategy versus conventional fixed-dose therapy. DESIGN: A prospective, randomized, 52 week, open-label trial of concentration-controlled compared with conventional dose zidovudine, lamivudine, and indinavir therapy conduced in a university-based general clinical research center in the United States. PATIENTS: Forty antiretroviral-naive individuals with plasma HIV-RNA levels > 5000 copies/ml. INTERVENTIONS: Zidovudine, lamivudine, and indinavir plasma concentrations were measured in all participants. Doses were adjusted in those assigned to concentration-controlled therapy to achieve levels equal to or greater than target values. MAIN OUTCOME MEASURES: The proportion of patients who achieved the desired drug concentrations, the proportion of patients with HIV-RNA levels < 50 copies/ml at week 52, and safety and tolerance in the concentration controlled versus conventional therapy arms. RESULTS: Significantly more concentration-controlled recipients achieved the desired concentration targets for all three drugs: 15 of 16 concentration-controlled recipients compared with nine of 17 conventional recipients (P = 0.017) had HIV-RNA levels < 50 copies/ml at week 52. No difference was observed in the occurrence of drug-related clinical events or laboratory abnormalities between the two treatment arms. CONCLUSION: Concentration-controlled therapy implemented simultaneously for three antiretroviral agents was feasible, as well tolerated as conventional therapy, and resulted in a greater proportion of recipients with HIV-RNA levels < 50 copies/ml after 52 weeks. These findings provide a scientific basis to challenge the accepted practice of administering the same dose of antiretroviral agents to all adults, ignoring the concentrations actually achieved. PMID- 11872999 TI - Final analysis of the Trilege induction-maintenance trial: results at 18 months. AB - BACKGROUND: First results of Trilege demonstrated that the strategy of less intensive antiviral therapy is less effective than continuation of triple-drug therapy. OBJECTIVE: To compare the final number of failures at month 18 and to study viral dynamics in patients experiencing a virological failure. DESIGN: Longitudinal follow-up from a randomized controlled trial. SETTING: Forty-three AIDS clinical-trial units. PATIENTS: A total of 279 HIV-1 infected adults randomized in Trilege. MEASUREMENTS: Analysis of recurrent values of HIV RNA > 500 copies/ml beyond time to virologic failure. RESULTS: A total of 83 patients experienced virological failure by month 18; 10 in the zidovudine (ZDV) + lamivudine (3TC) + indinavir (IDV) arm, 46 in the ZDV + 3TC arm, and 27 in the ZDV + IDV arm, confirming previous results. Whatever the treatment ultimately received, 87% of patients had an HIV RNA < 500 copies/ml at month 18 with no statistical difference between randomized arms. Patients experiencing a failure in the triple-drug regimen had a greater tendency to maintain HIV RNA > 500 copies/ml beyond the time of virological failure than patients in both less intensive treatment groups who experienced failure. Lower levels of HIV RNA at failure and reinitiating of either the original triple-drug regimen or a new combination of nucleoside analogue reverse transcriptase inhibitors and protease inhibitors were associated with lower hazard ratios for developing recurrent HIV RNA > 500 copies/ml. CONCLUSION: Results confirmed the failure of a less intensive regimen to maintain patients with a viral suppression (HIV RNA < 500 copies/ml). Although there is a lower incidence of failure in the triple-drug regimen, randomization to a less intensive regimen of ZDV + 3TC or ZDV + IDV was not detrimental, as treatment modification, either to the original triple regimen, or a different regimen was successful. PMID- 11873000 TI - Pharmacokinetic interactions between protease inhibitors and statins in HIV seronegative volunteers: ACTG Study A5047. AB - OBJECTIVE: Lipid lowering therapy is used increasingly in persons with HIV infection in the absence of safety data or information on drug interactions with antiretroviral agents. The primary objectives of this study were to examine the effects of ritonavir (RTV) plus saquinavir soft-gel (SQVsgc) capsules on the pharmacokinetics of pravastatin, simvastatin, and atorvastatin, and the effect of pravastatin on the pharmacokinetics of nelfinavir (NFV) in order to determine clinically important drug-drug interactions. DESIGN: Randomized, open-label study in healthy, HIV seronegative adults at AIDS Clinical Trials Units across the USA. METHODS: Three groups of subjects (arms 1, 2, and 3) received pravastatin, simvastatin or atorvastatin (40 mg daily each) from days 1-4 and 15-18. In these groups, RTV 400 mg and SQVsgc 400 mg twice daily were given from days 4-18. A fourth group (arm 4) received NFV 1250 mg twice daily from days 1-14 with pravastatin 40 mg daily added from days 15-18. Statin and NFV levels were measured by liquid chromatography/tandem mass spectrometry. RESULTS: Fifty-six subjects completed both pharmacokinetic study days. In arms 1-3, the median estimated area under the curves (AUC)(0-24) for the statins were: pravastatin (arm 1, n = 13), 151 and 75 ng.h/ml on days 4 and 18 (decline of 50% in presence of RTV/SQVsgc), respectively (P = 0.005); simvastatin (arm 2, n = 14), 17 and 548 ng.h/ml on days 4 and 18 (increase of 3059% in the presence of RTV/SQVsgc), respectively (P < 0.001); and total active atorvastatin (arm 3, n = 14), 167 and 289 ng.h/ml on days 4 and 18 (increase of 79% in the presence of RTV/SQVsgc), respectively (P < 0.001). In arm 4, the median estimated AUC(0-8) for NFV (24 319 versus 26 760 ng.h/ml; P = 0.58) and its active M8 metabolite (15 565 versus 14 571 ng.h/m; P = 0.63) were not statistically different from day 14 to day 18 (without or with pravastatin). CONCLUSIONS: Simvastatin should be avoided and atorvastatin may be used with caution in persons taking RTV and SQVsgc. Dose adjustment of pravastatin may be necessary with concomitant use of RTV and SQVsgc. Pravastatin does not alter the NFV pharmacokinetics, and thus appears to be safe for concomitant use. PMID- 11873001 TI - A randomized trial assessing the impact of phenotypic resistance testing on antiretroviral therapy. AB - OBJECTIVE: To compare the effect of treatment decisions guided by phenotypic resistance testing (PRT) or standard of care (SOC) on short-term virological response. DESIGN: A prospective, randomized, controlled clinical trial conducted in 25 university and private practice centers in the United States. PARTICIPANTS: A total of 272 subjects who failed to achieve or maintain virological suppression (HIV-1-RNA plasma level > 2000 copies/ml) with previous exposure to two or more nucleoside reverse transcriptase inhibitors and one protease inhibitor. INTERVENTIONS: Randomization was to antiretroviral therapy guided by PRT or SOC. MAIN OUTCOME MEASURES: The percentage of subjects with HIV-1-RNA plasma levels less than 400 copies/ml at week 16 (primary); change from baseline in HIV-1-RNA plasma levels and number of "active" (less than fourfold resistance) antiretroviral agents used (secondary). RESULTS: At week 16, using intent-to treat (ITT) analysis, a greater proportion of subjects had HIV-1-RNA levels less than 400 copies/ml in the PRT than in the SOC arm (P = 0.036, ITT observed; P = 0.079, ITT missing equals failure). An ITT observed analysis showed that subjects in the PRT arm had a significantly greater median reduction in HIV-1-RNA levels from baseline than the SOC arm (P = 0.005 for 400 copies/ml; P = 0.049 for 50 copies/ml assay detection limit). Significantly more subjects in the PRT arm were treated with two or more "active" antiretroviral agents than in the SOC arm (P = 0.003). CONCLUSION: Antiretroviral treatment guided prospectively by PRT led to the increased use of "active" antiretroviral agents and was associated with a significantly better virological response. PMID- 11873002 TI - Parallel decline of CD8+/CD38++ T cells and viraemia in response to quadruple highly active antiretroviral therapy in primary HIV infection. AB - OBJECTIVES: To monitor changes in the numbers of CD8 lymphocytes expressing the activated CD38++ phenotype in peripheral blood samples from patients with primary HIV infection (PHI) treated with highly active antiretroviral therapy (HAART). METHODS: Zidovudine, lamivudine, abacavir and amprenavir were initiated during PHI as part of the Quest study. Absolute numbers of CD8+/CD38++ T cells were determined using three-colour flow cytometry, and plasma viral load (VL) was measured using the Roche Amplicor method. RESULTS: The median, pre-therapy CD8+/CD38++ T cell count was 461/mm(3)(interquartile range 216, 974) in 131 patients compared with normal control values of less than 20 cells/mm(3). Levels fell markedly in parallel with VL within the first 2 weeks of HAART initiation, to a median of 47 cells/mm(3) at 28 weeks (median 436 cell decline; P < 0.001). At that time, 80% of patients had a VL less than 50 copies/ml, and 16.3% of all patients had less than 20 CD8+/CD38++ T cells/mm(3). A continued decrease in CD8+/CD38++ T cell count occurred in 67.2% of patients whose VL was maintained below 50 copies/ml (median change from first to last value -18 cells/mm(3); P < 0.001). CONCLUSION: After the initiation of HAART in PHI, CD8+/CD38++ lymphocytes declined rapidly in parallel with VL, and allowed for a normalization of CD8+/CD38++ T cell numbers in a subset of patients at week 28. Cell numbers continued to decline in patients who maintained VL below 50 copies/ml, indicating that the CD8+/CD38++ T cell count may represent a marker of residual viral replication when VL falls below detectable levels after HAART intervention. PMID- 11873003 TI - HIV-1 infection in rural Africa: is there a difference in median time to AIDS and survival compared with that in industrialized countries? AB - OBJECTIVES: To describe the progression times of HIV-1 infection from seroconversion to AIDS and to death, and time from first developing AIDS to death in rural Uganda. Also, to describe the proportion of individuals within the cohort dying with AIDS and the CD4 lymphocyte count before death. DESIGN: A prospective, longitudinal, population-based cohort. METHODS: Since 1990, 107 HIV prevalent cases, 168 incident cases and 235 HIV-seronegative controls have been recruited into a cohort in rural Uganda. Participants are recruited from the general population and they are reviewed routinely every 3 months and at other times when ill. RESULTS: The median time from seroconversion to death was 9.8 years. Age over 40 years at seroconversion was associated with more rapid progression (P < 0.001, log rank test). For the first 4 years of the study, HIV contributed little to the death rates in the HIV incident cases, but after 5 years, the contribution of HIV became greater and was particularly marked in the oldest age group. Survival rates in the cohort were similar to those in the general population. The median time from seroconversion to AIDS was 9.4 years and from AIDS to death was 9.2 months. Of those infected with HIV-1, 80% died with AIDS and 20% had a CD4 count < 10 x 106 cells/l. CONCLUSIONS: Survival with HIV-1 infection is similar in Africa to industrialized countries before the use of antiretroviral therapy; when they do die, many of those in Africa are severely immunosuppressed and most have clinical features of AIDS. PMID- 11873005 TI - Raised viral load in patients with viral suppression on highly active antiretroviral therapy: transient increase or treatment failure? AB - OBJECTIVE: To assess the occurrence of viral load greater than 50 copies/ml in patients on highly active antiretroviral therapy (HAART) having achieved less than 50 copies/ml and the chance of whether a viral load greater than 50 copies/ml would lead to a sustained and increasing viral load. DESIGN: A cohort of 553 patients on HAART with viral loads of less than 50 copies/ml were followed. RESULTS: Over a median of 56 weeks 35% of patients experienced a transient increase and 8% virological failure (two consecutive viral loads of > 400 copies/ml). Transient increases and virological failure were more common in those with greater drug experience, and those with initial raised viral load values of more than 400 copies/ml were more likely to have a sustained increase and become virological failures. CONCLUSION: Transient increases in viral load are common, mainly in the 50-400 copies/ml range, and the majority of subsequent viral load estimations show a return to less than 50 copies/ml. A single raised viral load should lead to adherence support and intensified monitoring. Subsequent treatment decisions can then be based on evidence of true virological rebound and failure. PMID- 11873004 TI - Validation of a simplified medication adherence questionnaire in a large cohort of HIV-infected patients: the GEEMA Study. AB - OBJECTIVE: To assess the effectiveness of the simplified medication adherence questionnaire (SMAQ) in identifying non-adherent patients. DESIGN: Prospective observational study of adherence. The six-item SMAQ was developed. The following aspects were evaluated: (i) criterion validity, comparison with electronic adherence monitoring; (ii) construct validity, association between adherence, as defined by the SMAQ, and virological outcomes; and (iii) reliability, internal consistency and reproducibility. PATIENTS: A group of 3004 unselected HIV patients who had initiated nelfinavir therapy combined with other antiretroviral drugs [21% naive, 15% protease inhibitor (PI)-naive, 64% PI-experienced] between January 1998 and December 1999 were enrolled in 69 hospitals in Spain. The SMAQ was administered at months 3, 6 and 12. RESULTS: The SMAQ showed 72% sensitivity, 91% specificity and a likelihood ratio of 7.94 to identified non-adherent patients, compared with the medication-event monitoring system (40 patients evaluated). At month 12, 1797 patients were evaluated, of whom 32.3% were defined as non-adherent; viral load < 500 copies/ml found in 68.3% of the adherent, and 46% of the non-adherent patients. A logistic regression analysis of PI-naive patients was performed, including age, sex, baseline viral load > 5 log10/ml, CD4 cell count < 200 x 10(6)/l, and non-adherence as independent variables. Non adherence was the only significant risk factor in failing to achieve virological suppression. Cronbach's alpha internal consistency coefficient was 0.75, and overall inter-observer agreement was 88.2%. CONCLUSION: The SMAQ appears to be an adequate instrument with which to assess adherence in HIV-infected patients, and may be applied in most clinical settings. PMID- 11873006 TI - Low prevalence of primary mutations associated with drug resistance in antiviral naive patients at therapy initiation. AB - OBJECTIVE: To assess the prevalence of mutations in the reverse-transcriptase (RT) and protease (PR) region in a cohort of chronically-infected HIV-positive patients requiring highly active antiretroviral therapy (HAART). METHODS: The study included 347 patients enrolled in the Italian Cohort of Antiretroviral Naive patients (I.CO.NA) who had to initiate HAART. The whole PR-region, and amino acids 1-320 of RT-region were sequenced from plasma samples at baseline. RESULTS: Median CD4-lymphocytes and HIV-RNA at baseline were 231 x 10(6) cells/l and 4.89 log(10) copies/ml; 307 of 347 (88.5%) patients carried no mutations in the RT region, whereas 40 (11.5%) carried one or more mutations associated with resistance to nucleoside-RT inhibitor (NRTI) (7.8%), or non-nucleoside-RTI (NNRTI) (4.9%), with four patients carrying mutations to both classes. Among mutations associated with high-level resistance to RTI, T215Y was found in only two patients, M184V in two cases, T69D and T215C in other two cases (one each), and K103N in only one patient, for a total of six patients (one carrying both T215Y and M184V) (1.7%). Seventy-six patients (21.9%) carried no mutations in the PR region, whereas 271 (78.1%) had one or more mutations. Primary mutations associated with substantial resistance to protease inhibitors were found in only five of 347 patients (1.4%) (M46V/L, I54V, V82A/I); all the other patients carried only secondary mutations (L10F/I/V, M36I, L63P, A71T/V, V77I). CONCLUSIONS: Prevalence of mutations associated with high-level resistance to antiretroviral drugs is low in HIV-infected patients with long-term infection. This suggests no preclusion in principle to any antiretroviral drug at the time of decision of the first therapeutic regimen. PMID- 11873007 TI - Changes in plasma HIV-1-RNA viral load and CD4 cell counts, and lack of zidovudine resistance among pregnant women receiving short-course zidovudine. AB - OBJECTIVE: To describe changes in HIV-1 plasma viral load (VL) and CD4 cell counts and to assess zidovudine resistance associated with a short course of oral zidovudine during late pregnancy. METHODS: From April 1996 to February 1998 in Abidjan, Cote d'Ivoire, 280 HIV-1-seropositive women were randomly assigned at 36 weeks' gestation to receive zidovudine (300 mg) or placebo twice a day, and then one tablet every 3 h from the onset of labor until delivery. Blood samples obtained every 2 weeks until delivery, then at 2 and 4 weeks, and 3 or 6 months after delivery were tested from selected women based on duration of therapy for plasma VL and CD4 cell counts, and samples from 20 women in the zidovudine group were tested by DNA sequencing for the presence of zidovudine resistance mutations. RESULTS: In the zidovudine group, the median reduction in plasma VL (log(10) copies/ml) was -0.48 after 2 weeks (P = 0.02 versus placebo), -0.48 after 4 weeks (P = 0.06), -0.80 after 6 weeks (P = 0.29) of treatment, -0.12 at delivery (P = 0.11), +0.21 at 2 weeks (P = 0.83), +0.17 at 4 weeks (P = 0.69), and +0.21 at 3 months (P = 0.56) postpartum. Median CD4 cell counts were higher in the zidovudine than in the placebo group after 2, 4, and 6 weeks of treatment (P < 0.05). No mutations associated with zidovudine resistance were identified in any of the samples tested. CONCLUSION: These findings suggest that a short course of zidovudine has no adverse HIV-1 virological consequences for the mother. PMID- 11873008 TI - Twenty-four month efficacy of a maternal short-course zidovudine regimen to prevent mother-to-child transmission of HIV-1 in West Africa. AB - OBJECTIVE: To assess the 24 month efficacy of a maternal short-course zidovudine regimen to prevent mother-to-child transmission (MTCT) of HIV-1 in a breastfeeding population in West Africa. METHODS: Data were pooled from two clinical trials: DITRAME-ANRS049a conducted in Abidjan, Cote d'Ivoire and Bobo Dioulasso, Burkina-Faso and RETRO-CI, conducted in Abidjan. Between September 1995 and February 1998, consenting HIV-1-seropositive women were randomly assigned to receive zidovudine (300 mg) or placebo: one tablet twice daily from 36-38 weeks' gestation until delivery, then in DITRAME only, for 7 more days. Paediatric HIV-1 infection was defined as a positive HIV-1 polymerase chain reaction, or if aged > or =15 months, a positive HIV-1 serology. Cumulative risks (CR) of infection were estimated using a competing risk approach with weaning as a competing event. RESULTS: Among 662 live-born children, 641 had at least one HIV-1 test. All but 12 children were breastfed. At 24 months, overall CR of MTCT were 0.225 in the zidovudine and 0.302 in the placebo group, a 26% significant reduction. Among children born to women with CD4 cell counts < 500/ml at enrollment, CR of MTCT were similar, 0.396 in the zidovudine and 0.413 in the placebo group. Among children born to women with CD4 cell counts > or =500/ml, CR of MTCT were 0.091 in the zidovudine and 0.220 in the placebo group, a significant 59% reduction. CONCLUSION: A maternal short-course zidovudine regimen reduces MTCT of HIV-1 at age 24 months, despite prolonged breastfeeding. However, efficacy was observed only among women with CD4 cell counts > or =500/ml. New interventions should be considered to prevent MTCT, especially for African women with advanced HIV-1 immunodeficiency. PMID- 11873009 TI - Study of bias in antenatal clinic HIV-1 surveillance data in a high contraceptive prevalence population in sub-Saharan Africa. AB - OBJECTIVE: To describe patterns, sources and consequences of bias in antenatal clinic (ANC) HIV prevalence estimates in a high contraceptive prevalence population. BACKGROUND: HIV surveillance in Africa relies on data from pregnant women attending ANCs. HIV estimates from pregnant women understate female infection levels in low income, high fertility populations. Bias in high contraceptive use, delayed sexual debut populations remains undescribed. DESIGN AND METHOD: Comparison of parallel cross-sectional population and antenatal survey data from rural Zimbabwe, where 60% of women are recent contraceptive users. RESULTS: HIV prevalence in recently pregnant women (25.7%; n = 576) and all women (25.5%; n = 5138) is similar over the age-range 15-44 years. As in high fertility populations, HIV prevalence is higher in pregnant women at young ages and lower at older ages but the crossover point occurs later due to delayed sexual activity. HIV understatement at older ages due to HIV-associated infertility is mitigated by less HIV infection and less frequent ANC attendance in contraceptive users. The local ANC HIV prevalence estimate is lower [21.2%; n = 1215; risk ratio versus pregnant women in the general population, 0.8; 95% confidence interval (CI), 0.7-1.0], possibly because women from more remote areas are included. ANC estimates overstate the relative risk of HIV in more educated women (age-adjusted odds ratio, 1.1; 95% CI, 0.8-1.4 versus 0.7; 95% CI, 0.6 0.9). CONCLUSIONS: ANC estimates understate female HIV prevalence in this low fertility population but, here, the primary cause is not selection of pregnant women. ANC estimate adjustment procedures that control for contraceptive use and age at first sex are needed. PMID- 11873010 TI - "Barebacking" in a diverse sample of men who have sex with men. AB - OBJECTIVES: To assess the prevalence of and factors associated with "Barebacking" as a sociocultural phenomenon in a sample of HIV-positive and -negative men who have sex with men (MSM), and to assess the reasons for barebacking and venues for meeting partners. DESIGN: A cross-sectional survey of MSM recruited in the San Francisco Bay Area from July 2000 to February 2001. METHODS: Barebacking, defined as "intentional anal sex without a condom with someone other than a primary partner", was assessed among men who had heard of the term. Participants were recruited outside multiple venues and interviewed later at community locations. Chi-square and multivariate logistic regression were used for analysis. RESULTS: The sample (n = 554) of MSM were African-American (28%), Latino (27%), white (31%) and other race/ethnicity (14%); 35% reported being HIV-positive. Seventy per cent of the men had heard of barebacking. Among men aware of the term, 14% had barebacked in the past 2 years (22% of HIV-positive versus 10% of HIV negative men, P < 0.001); 10% of the full sample did so. The prevalence of barebacking did not differ by race/ethnicity or sexual orientation identification. Men tended to report bareback partners who had the same HIV serostatus; however, a sizeable proportion of men had partners of different or unknown serostatus. Increased physical stimulation and emotional connectedness were the primary reasons for barebacking. CONCLUSION: New approaches are needed to reduce bareback behavior and the risk of HIV transmission, including innovative health-promoting behavioral and biomedical interventions. PMID- 11873011 TI - Comparison of HIV prevalences in community-based and antenatal clinic surveys in rural Mwanza, Tanzania. AB - OBJECTIVES: First, to compare the prevalence of HIV infection among women in the general population and antenatal clinic (ANC) attenders in rural Mwanza, Tanzania, and second, to validate a method for adjusting HIV prevalence in ANC attenders to estimate the prevalence in the general female population aged 15-44 years. METHODS: A cross-sectional population survey was conducted in 12 rural communities of Mwanza Region between 1991 and 1992. From the same communities sequential ANC attenders were recruited on two occasions between 1991 and 1993. Consenting subjects were interviewed, examined, treated and a serum sample was tested for HIV. The HIV prevalence in women in the general population was compared with unadjusted and adjusted prevalences in ANC attenders. Parity adjusted prevalences were obtained by applying correction factors to the observed prevalences in parous and nulliparous ANC attenders. RESULTS: A total of 5675 women aged 15-44 years from the general population and 2265 ANC attenders had complete socio-demographic and laboratory data. Unadjusted HIV prevalence was significantly lower in ANC attenders (3.6%) than women from the general population (4.7%, P = 0.025), but after adjustment there was no significant difference between the two groups (4.6 versus 4.7%, P = 0.95). CONCLUSION: In this rural population, the HIV prevalence in ANC attenders underestimated the prevalence among women in the general population, but this difference was eliminated by applying parity-based correction factors. Information on parity should be routinely collected in ANC-based HIV sentinel surveillance. PMID- 11873012 TI - Anti-idiotypic antibody Ab2/3H6 mimics the epitope of the neutralizing anti-HIV-1 monoclonal antibody 2F5. AB - Anti-idiotypic antibodies directed against potentially neutralizing anti-HIV-1 antibodies may mimic epitopes on gp41 otherwise cryptic to the immune system. This study reports the generation of murine monoclonal antibody Ab2/3H6 blocking the binding of human Ab1 2F5 to the synthetic epitope and to gp160 in an enzyme linked immunosorbent competition assay. Ab2/2H6 diminished the neutralizing potency of 2F5 in an in-vitro neutralization assay. Ab2/3H6 Fab fragments were capable of inducing neutralizing immune and 2F5-specific responses in B6D2F1 mice applying a simple prime-boost regimen of immunization. PMID- 11873013 TI - Biological characteristics of newly described HIV-1 BG recombinants in Spanish individuals. PMID- 11873014 TI - Discontinuation of secondary prophylaxis for penicilliosis marneffei in AIDS patients responding to highly active antiretroviral therapy. PMID- 11873015 TI - Inflammatory oedema associated with lopinavir-including HAART regimens in advanced HIV-1 infection: report of 3 cases. PMID- 11873016 TI - Poor solubility limiting significance of in-vitro studies with HIV protease inhibitors. PMID- 11873017 TI - First, do no harm: a call for emphasizing adherence and HIV prevention interventions in active antiretroviral therapy programs in the developing world. PMID- 11873018 TI - Concurrency and sexual transmission. PMID- 11873021 TI - Measurement of pain in children: state-of-the-art considerations. PMID- 11873022 TI - No preemptive analgesia: is that so bad? PMID- 11873023 TI - Validation of the Non-communicating Children's Pain Checklist-Postoperative Version. AB - BACKGROUND: This study evaluated the psychometric properties of the Non communicating Children's Pain Checklist-Postoperative Version (NCCPC-PV) when used with children with severe intellectual disabilities. METHODS: The caregivers of 24 children with severe intellectual disabilities (aged 3-19 yr) took part. Each child was observed by one of their caregivers and one of the researchers for 10 min before and after surgery. They independently completed the NCCPC-PV and made a visual analog scale rating of the child's pain intensity for those times. A nurse also completed a visual analog scale for the same observations. RESULTS: The NCCPC-PV was internally reliable (Cronbach alpha = 0.91) and showed good interrater reliability. A repeated-measures analysis of variance indicated NCCPC PV total and subscale scores were significantly higher after surgery and did not differ by observer. Postoperative NCCPC-PV scores correlated with visual analog scale ratings provided by caregivers and researchers, but not with those of nurses. A score of 11 on the NCCPC-PV, by caregivers, provided 0.88 sensitivity and 0.81 specificity for classifying children with moderate to severe pain. CONCLUSIONS: The NCCPC-PV displayed good psychometric properties when used for the postoperative pain of children with severe intellectual disabilities and has the potential to be useful in a clinical setting. The results suggest familiarity with an individual child with intellectual disabilities is not necessary for pain assessment. PMID- 11873024 TI - Lung function under high thoracic segmental epidural anesthesia with ropivacaine or bupivacaine in patients with severe obstructive pulmonary disease undergoing breast surgery. AB - BACKGROUND: Because general anesthesia with tracheal intubation can elicit life threatening bronchospasm in patients with bronchial hyperreactivity, epidural anesthesia is often preferred. However, segmental high thoracic epidural anesthesia (sTEA) causes pulmonary sympathetic and respiratory motor blockade. Whether it can be safely used for chest wall surgery as a primary anesthetic technique in patients with chronic obstructive pulmonary disease or asthma is unclear. Furthermore, ropivacaine supposedly evokes less motor blockade than bupivacaine and might minimize side effects. To test the feasibility of the technique and the hypotheses that (1) sTEA with ropivacaine or bupivacaine does not change lung function and (2) there is no difference between sTEA with ropivacaine or bupivacaine, the authors studied 20 patients with severe chronic obstructive pulmonary disease (forced expiratory volume in 1 s [FEV1] = 52.1 +/- 17.3% of predicted [mean +/- SD]) or asthma who were undergoing breast surgery. METHODS: In a double-blind, randomized fashion, sTEA was performed with 6.6 +/- 0.5 ml of either ropivacaine, 0.75% (n = 10), or bupivacaine, 0.75% (n = 10). FEV1, vital capacity, FEV1 over vital capacity, spread of analgesia (pin prick), hand and foot skin temperatures, mean arterial pressure, heart rate, and local anesthetic plasma concentrations were measured with patients in the sitting and supine positions before and during sTEA. RESULTS: Segmental high thoracic epidural anesthesia (segmental spread C4-T8 [bupivacaine] and C5-T9 [ropivacaine]) significantly decreased FEV1 from 1.22 +/- 0.54 l (supine) to 1.09 +/- 0.56 l (ropivacaine) and from 1.23 +/- 0.49 l to 1.12 +/- 0.46 l (bupivacaine). In contrast, FEV1 over vital capacity increased from 64.6 +/- 13.5 to 68.2 +/- 14.5% (ropivacaine) and from 62.8 +/- 12.4 to 66.5 +/- 13.6% (bupivacaine). There was no difference between ropivacaine and bupivacaine. Skin temperatures increased significantly, whereas arterial pressure and heart rate significantly decreased indicating widespread sympathetic blockade. All 20 patients tolerated surgery well. CONCLUSIONS: Despite sympathetic blockade, sTEA does not increase airway obstruction and evokes only a small decrease in FEV1 as a sign of mild respiratory motor blockade with no difference between ropivacaine and bupivacaine. Therefore, sTEA can be used in patients with severe chronic obstructive pulmonary disease and asthma undergoing chest wall surgery as an alternative technique to general anesthesia. PMID- 11873025 TI - Chronobiology of epidural ropivacaine: variations in the duration of action related to the hour of administration. AB - BACKGROUND: A temporal pattern of the kinetics of local anesthetics is demonstrated in dental and skin anesthesia, with an important variation in the duration of action related to the hour of administration. The aim of this study is to determine whether the hour of injection influences the duration of epidurally administered ropivacaine during labor. METHODS: One hundred ninety four women in the first stage of labor were assigned to one of four groups throughout the day period: group 1 (night: from 1:01 to 7:00 am), group 2 (morning: from 7:01 am to 1:00 pm), group 3 (afternoon: from 1:01 to 7:00 pm), and group 4 (evening: from 7:01 pm to 1:00 am). Each patient received 14 ml ropivacaine, 0.17%, epidurally, and analgesia duration was measured. RESULTS: Pain assessed by a visual analog score was not differ-ent among groups before the first injection of local anesthetic. Analgesia duration was greater in the diurnal period (group 2: 110 +/- 25 min and group 3: 117 +/- 23 min) compared with the nocturnal period (group 1: 94 +/- 23 min and group 4: 91 +/- 23 min) (P < 0.01). The largest intraday variation of analgesia duration among groups reached 28%. CONCLUSIONS: Epidural analgesia duration exhibits a temporal pattern with important differences among diurnal and nocturnal phases. The authors emphasize that the lack of consideration of the chronobiologic conditions in epidural analgesia studies may create significant statistical bias. Future studies dealing with epidural local anesthetics should consider the time of drug administration. PMID- 11873026 TI - Cesarean delivery: a randomized trial of epidural analgesia versus intravenous meperidine analgesia during labor in nulliparous women. AB - BACKGROUND: Controversy concerning increased cesarean births as a result of epidural analgesia for relief of labor pain has been attributed, in large part, to difficulties interpreting published studies because of design flaws. In this study, the authors compared epidural analgesia to intravenous meperidine analgesia using patient-controlled devices during labor to evaluate the effects of labor epidural analgesia, primarily on the rate of cesarean deliveries while minimizing limitations attributable to study design. METHODS: Four hundred fifty nine nulliparous women in spontaneous labor at term were randomly assigned to receive either epidural analgesia or intravenous meperidine analgesia. Epidural analgesia was initiated with 0.25% bupivacaine and was maintained with 0.0625% bupivacaine and fentanyl 2 microg/ml at 6 ml/h with 5-ml bolus doses every 15 min as needed using a patient-controlled pump. Women in the intravenous analgesia group received 50 mg meperidine with 25 mg promethazine hydrochloride as an initial bolus, followed by 15 mg meperidine every 10 min as needed, using a patient-controlled pump. A written procedural manual that prescribed the intrapartum obstetric management was followed for each woman randomized in the study. RESULTS: A total of 226 women were randomized to receive epidural analgesia, and 233 women were randomized to receive intravenous meperidine analgesia. Protocol violations occurred in 8% (38 of 459) of women. There was no difference in the rate of cesarean deliveries between the two analgesia groups (epidural analgesia, 7% [16 of 226; 95% confidence interval, 4-11%] vs. intravenous meperidine analgesia, 9% [20 of 233; 95% confidence interval, 5-13%]; P = 0.61). Significantly more women randomized to epidural analgesia had forceps deliveries compared with those randomized to meperidine analgesia (12% [26 of 226] vs. 3% [7 of 233]; P < 0.001). Women who received epidural analgesia reported lower pain scores during labor and delivery compared with women who received intravenous meperidine analgesia. CONCLUSIONS: Epidural analgesia compared with intravenous meperidine analgesia during labor does not increase cesarean deliveries in nulliparous women. PMID- 11873027 TI - Inability to consistently elicit a motor response following sensory paresthesia during interscalene block administration. AB - BACKGROUND: Two methods of nerve block based on eliciting neural feedback with the block needle currently exist. The paresthesia technique uses sensory feedback to ascertain that the needle tip is close to the nerve. By contrast, a peripheral nerve stimulator makes use of motor responses to electrical stimulation. The relation of motor responses to an electrical peripheral nerve stimulator and sensory nerve contact (paresthesia) had not been studied. METHODS: Thirty consecutive unpremedicated patients who presented for shoulder surgery with interscalene block anesthesia were prospectively studied. Interscalene block was performed by the single paresthesia method of Winnie, using an insulated or non insulated needle connected to a peripheral nerve stimulator with the power off. At the precise point of paresthesia, the peripheral nerve stimulator was turned on, and the current was slowly increased to 1.0 mA with a pulse width of 0.2 ms. Presence and location of any motor responses were observed and recorded. RESULTS: All patients had easily elicited paresthesias. The site of first paresthesia was to the shoulder in 73% of patients. Only 30% of patients exhibited any motor response to electrical stimulation up to 1.0 mA. There was no relation between site of paresthesia and associated motor nerve response. CONCLUSION: Elicitation of paresthesia does not translate to an ability to elicit a motor response to a peripheral nerve stimulator in the majority of patients. PMID- 11873028 TI - The influence of remifentanil on the dynamic relationship between sevoflurane and surrogate anesthetic effect measures derived from the EEG. AB - BACKGROUND: The authors modeled the influence of remifentanil on the dynamics of sevoflurane using three parameters derived from the electroencephalogram: 95% spectral edge frequency (SEF), canonical univariate parameter (CUP), and Bispectral Index (BIS). METHODS: Thirty-six patients with American Society of Anesthesiologists physical status class I or II were recruited, of which 12 received a target remifentanil concentration of 0 ng/ml, eight 2 ng/ml, eight 4 ng/ml, and another eight 8 ng/ml. Next (before surgery), several step-wise changes in the end-tidal sevoflurane concentration (F(ET,sevo)) were performed. A data acquisition system simultaneously recorded F(ET,sevo), the raw electroencephalogram, BIS, and SEF. The authors used a combination of an effect compartment and an inhibitory sigmoid E(MAX) model to describe the relation between F(ET,sevo) and BIS, SEF, and CUP. Model parameters (t(1/2)k(e0), E(MAX), E(MIN), C(50), gamma, CUP weight factors) were estimated using the population data analysis program NONMEM. Significant remifentanil model parameter dependencies (P < 0.01) were determined. RESULTS: Determined from SEF, remifentanil had no effect on t(1/2)k(e0) (1.91 +/- 0.26 min [mean +/- standard error]) but caused an increase in C(50) (baseline = 1.48 +/- 0.12%; 80% increase at 8 ng/ml) and decrease in E(MIN) (baseline = 10.8 +/- 0.6 Hz; 80% reduction at 8 ng/ml). Determined from CUP, remifentanil caused a dose-dependent decrease in t(1/2)k(e0) (baseline = 4.31 +/- 1.00 min; 60% decrease at 8 ng/ml), with no effect on C(50) (baseline = 0.88 +/- 0.13%). Determined from BIS, remifentanil caused a dose-dependent decrease in t(1/2)k(e0) (baseline value = 3.11 +/- 0.32 min; 40% decrease at 8 ng/ml), without affecting C(50) (baseline = 1.12 +/- 0.05%). Median R(2) values of the pooled data set were 0.815 for SEF, 0.933 for CUP (P < 0.01 vs. SEF), and 0.952 for BIS (P < 0.01 vs. SEF and CUP). Addition of remifentanil increased the R(2) values for CUP only. CONCLUSIONS: Remifentanil accelerates sevoflurane blood-brain equilibration without affecting its hypnotic potency as determined from BIS and CUP. In terms of R(2), the authors' pharmacodynamic model describes the anesthetic-BIS relation best. PMID- 11873029 TI - Development and evaluation of a graphical anesthesia drug display. AB - BACKGROUND: Usable real-time displays of intravenous anesthetic concentrations and effects could significantly enhance intraoperative clinical decision-making. Pharmacokinetic models are available to estimate past, present, and future drug effect-site concentrations, and pharmacodynamic models are available to predict the drug's associated physiologic effects. METHODS: An interdisciplinary research team (bioengineering, architecture, anesthesiology, computer engineering, and cognitive psychology) developed a graphic display that presents the real-time effect-site concentrations, normalized to the drugs' EC(95), of intravenous drugs. Graphical metaphors were created to show the drugs' pharmacodynamics. To evaluate the effect of the display on the management of total intravenous anesthesia, 15 anesthesiologists participated in a computer-based simulation study. The participants cared for patients during two experimental conditions: with and without the drug display. RESULTS: With the drug display, clinicians administered more bolus doses of remifentanil during anesthesia maintenance. There was a significantly lower variation in the predicted effect-site concentrations for remifentanil and propofol, and effect-site concentrations were maintained closer to the drugs' EC(95). There was no significant difference in the simulated patient heart rate and blood pressure with respect to experimental condition. The perceived performance for the participants was increased with the drug display, whereas mental demand, effort, and frustration level were reduced. In a post-simulation questionnaire, participants rated the display to be a useful addition to anesthesia monitoring. CONCLUSIONS: The drug display altered simulated clinical practice. These results, which will inform the next iteration of designs and evaluations, suggest promise for this approach to drug data visualization. PMID- 11873030 TI - Beneficial effects of short-term vasopressin infusion during severe septic shock. AB - BACKGROUND: Septic shock is associated with vasopressin deficiency and a hypersensitivity to its exogenous administration. The goal of the current study was to determine whether short-term vasopressin infusion in patients experiencing severe septic shock has a vasopressor sparing effect while maintaining hemodynamic stability and adequate end-organ perfusion. METHODS: Patients experiencing septic shock that required high-dose vasopressor support were randomized to a double-blinded 4-h infusion of either norepinephrine (n = 11) or vasopressin (n = 13), and open-label vasopressors were titrated to maintain blood pressure. To assess end-organ perfusion, urine output and creatinine clearance, gastric mucosal carbon dioxide tension, and electrocardiogram ST segment position were measured. RESULTS: Patients randomized to norepinephrine went from a median prestudy norepinephrine infusion of 20.0 microg/min to a blinded infusion of 17.0 mug/min at 4 h, whereas those randomized to vasopressin went from a median prestudy norepinephrine infusion of 25.0 microg/min to 5.3 microg/min at 4 h (P < 0.001). Mean arterial pressure and cardiac index were maintained in both groups. Urine output did not change in the norepinephrine group (median, 25 to 15 ml/h) but increased substantially in the vasopressin group (median, 32.5 to 65 ml/h; P < 0.05). Similarly, creatinine clearance did not change in the norepinephrine group but increased by 75% in the vasopressin group (P < 0.05). Gastric mucosal carbon dioxide tension and electrocardiogram ST segments did not change significantly in either group. CONCLUSIONS: The authors conclude that short-term vasopressin infusion spared conventional vasopressor use and improved some measures of renal function in patients with severe septic shock. PMID- 11873031 TI - The relationship between acceleromyographic train-of-four fade and single twitch depression. AB - BACKGROUND: During offset of nondepolarizing neuromuscular block, a train-of-four (TOF) fade ratio of 0.70 or greater is considered to reliably indicate the return of single twitch height (T1) to its control value. Studies using mechanomyography or electromyography confirm this observation. The authors' impressions when using the acceleromyograph as a neuromuscular monitor did not support these results. Therefore, the authors studied the relation between T1 and the TOF ratio (when measured by acceleromyography) during recovery from neuromuscular block. METHODS: Sixteen adult patients were studied. Anesthesia was induced with intravenous opioid plus 2.0-2.5 mg/kg propofol. Laryngeal mask placement or tracheal intubation was accomplished without the use of muscle relaxants. Anesthesia was maintained with nitrous oxide, desflurane (2.0-3.0%, end-tidal), and fentanyl. The response of the thumb to ulnar nerve stimulation was recorded with the TOF Guard acceleromyograph (Organon Teknika BV, Boxtel, The Netherlands). TOFs were administered every 15 s. After final calibration, 0.15 mg/kg mivacurium was administered. No further relaxants were administered. T1 and the TOF ratio were then recorded until the TOF ratio had returned to its initial value (+/- 5%). RESULTS: At a TOF ratio of 0.70 (during recovery of neuromuscular function), T1 averaged only 69 +/- 8% of control. At a TOF ratio of 0.90, T1 averaged 86 +/- 5% of control. To achieve 90% recovery of T1, a TOF ratio of 0.93 +/- 0.08 was required. CONCLUSION: Assumptions regarding the relation between T1 and the TOF ratio derived from studies using mechanomyography and electromyography do not necessarily apply to observations obtained using acceleromyography. PMID- 11873032 TI - Implicit memory for words played during isoflurane- or propofol-based anesthesia: the lexical decision task. AB - BACKGROUND: Unconscious processing of words during general anesthesia has been suggested after surgery with several tests of implicit memory. Patients can neither recall those words nor do they have explicit memories of other intraoperative events. It is unclear to what degree information is processed during general anesthesia and which tests are best suited to detect implicit memory. In the current study, a lexical decision paradigm not previously used to demonstrate implicit memory during anesthesia was used. METHODS: Sixty patients undergoing lumbar disc surgery were assigned to receive isoflurane infusion- or propofol infusion-based anesthesia combined with alfentanil infusions and a nitrous oxide-oxygen mixture. A control group of 10 medical students listened to tapes without receiving anesthesia. Two tapes, each containing a list of 30 low frequency German nouns repeated for 15 min, were prepared, with half of the patients listening to tape A and the other half listening to tape B during the operation. Exposure time was 15 min from the time of skin incision onward. In the test phase, approximately 7 h later, words from lists A and B plus 60 nonwords were presented in random order by a computer program. Subjects were asked to indicate, by pressing one of two response buttons, whether the spoken word was or was not a legal German word (lexical decision). RESULTS: A recognition test revealed chance recognition for words presented during anesthesia. Lexical decision responses, however, were slightly faster to primed (previously presented) words than to unprimed (not previously presented) words when the entire group of patients was tested, suggesting a small implicit memory effect, which barely failed to reach the significance level. When the two medication groups were tested separately, no significant implicit memory effect could be ascertained statistically. The effects of previous exposure were much more pronounced in the control group. CONCLUSIONS: Balanced anesthesia techniques with isoflurane or propofol lead to only a minimal, statistically borderline implicit memory effect in the lexical decision paradigm. PMID- 11873033 TI - Relationship between intracranial pressure and critical closing pressure in patients with neurotrauma. AB - BACKGROUND: The driving pressure gradient for cerebral perfusion is the difference between mean arterial pressure (MAP) and critical closing pressure (CCP = zero flow pressure). Therefore, determination of the difference between MAP and CCP should provide an appropriate monitoring of the effective cerebral perfusion pressure (CPP(eff)). Based on this concept, the authors compared conventional measurements of cerebral perfusion pressure by MAP and intracranial pressure (CPP(ICP)) with CPP(eff). METHODS: Simultaneous synchronized recordings of pressure waveforms of the radial artery and blood flow velocities of the middle cerebral artery were performed in 70 head trauma patients. CCP was calculated from pressure-flow velocity plots by linear extrapolation to zero flow. RESULTS: Intracranial pressure measured by intraventricular probes and CCP ranged from 3 to 71 and 4 to 70 mmHg, respectively. Linear correlation between ICP and CCP was r = 0.91. CPP(ICP) was 77 +/- 20 mmHg and did not differ from CPP(eff); linear correlation was r = 0.92. However, limits of agreement were only +/- 16.2 mmHg. Therefore, in 51.4% of the patients, CPP(ICP) overestimated CPP(eff) by 19.8 mmHg at most. CONCLUSION: Assuming that CPP(eff) (MAP - CCP) takes into account more determinants of cerebral downstream pressure, in individual cases, the actual gold standard of CPP determination (MAP - ICP) might overestimate the CPP(eff) of therapeutic significance. PMID- 11873034 TI - Impact of shorter-acting neuromuscular blocking agents on fast-track recovery of the cardiac surgical patient. AB - BACKGROUND: Residual paralysis associated with the use of long-acting muscle relaxants can delay recovery from anesthesia and surgery. The authors tested the hypothesis that use of shorter-acting neuromuscular blocking agents is associated with reductions in tracheal extubation times and intensive care unit (ICU) length of stay in patients undergoing cardiac surgery with cardiopulmonary bypass. METHODS: One hundred ten patients scheduled for elective coronary artery bypass grafting or single valve surgery were randomized prospectively to receive either pancuronium or rocuronium intraoperatively. Anesthetic management and muscle relaxant maintenance dosing were standardized. In the ICU, the time required to wean ventilatory support, the duration of tracheal intubation, and length of stay were recorded. Subjects were asked to quantify generalized muscle weakness as they awakened in the ICU and again after tracheal extubation. RESULTS: Complete data were collected on 51 patients in the pancuronium group and 52 patients in the rocuronium group. No differences were found between the groups in anesthetic, surgical, or ICU management. Significant increases in the duration of weaning of ventilatory support were observed in patients who received pancuronium (median, 180 min; range, 50-780 min) compared with the rocuronium group (median, 110 min; range, 45-250 min). Tracheal extubation was significantly delayed in the pancuronium group (median, 500 min; range, 240-1,305 min) compared with the rocuronium group (median, 350 min; range, 210-1,140 min). Subjects in the pancuronium group experienced more mild to severe weakness in the ICU. However, the choice of muscle relaxant did not influence ICU length of stay. CONCLUSION: The use of shorter-acting neuromuscular blocking agents in patients undergoing cardiac surgery with cardiopulmonary bypass is associated with reductions in tracheal extubation times and symptoms of residual paresis. PMID- 11873035 TI - Morphologic changes in the upper airway of children during awakening from propofol administration. AB - BACKGROUND: The purpose of this study was to determine the morphologic changes that occur in the upper airway of children during awakening from propofol sedation. METHODS: Children undergoing magnetic resonance imaging of the head underwent additional scans of the upper airway during deep sedation with propofol; this was repeated on awakening. Axial views were obtained at the most posterior sites of the pharynx at the levels of the soft palate and tongue. Measurements were then obtained of the anterior-posterior (A-P) diameter, transverse diameter, and cross-sectional areas at these levels. RESULTS: Data were obtained on 16 children, aged 10 months to 7 yr. In both sedated and awakening states, most children had the smallest cross-sectional area of the pharynx at the level of the soft palate. During the sedated state, at the soft palate level, the transverse diameter was most narrow in 11 children, the A-P diameter was most narrow in 1 child, and they were equal in 2 children. During the sedated state, at the level of the tongue, the transverse diameter was most narrow in 9 children, the A-P diameter was most narrow in 5 children, and they were equal in 2 children. During awakening, at the soft palate level, the transverse diameter was most narrow in none of the children, the A-P diameter was most narrow in 13 children, and they were equal in 1 child. At the level of the tongue, the transverse diameter was most narrow in 4 children, and the A-P diameter was most narrow in 12 children. During awakening, the A-P diameter of the pharynx at the level of the soft palate decreased in 12 children, increased in 1 child, and remained the same in 1 child. (P < 0.001). The transverse diameter increased in 11 children, decreased in 1 child, and remained the same in 2 children (P = 0.001). The cross-sectional area at the level of the soft palate increased in 4 children, decreased in 8 children, and stayed the same in 2 children (P = 0.5). During awakening, the A-P diameter of the pharynx at the level of the tongue decreased in 11 children, increased in 4 children, and remained the same in 1 child. (P = 0.01). The transverse diameter increased in 11 children and decreased in 5 children (P = 0.07). The cross-sectional area at the level of the tongue increased in 7 children, decreased in 7 children, and stayed the same in 2 children (P = 0.9). CONCLUSIONS: The dimensions of the upper airways of children change shape significantly on awakening from propofol sedation. When sedated, the upper airway is oblong shaped, with the A-P diameter larger than the transverse diameter. On awakening, the shape of the upper airway in most children changed such that the transverse diameter was larger. Cross sectional areas between sedated and awakening states were unchanged. These changes may reflect the differential effects of propofol on upper airway musculature during awakening. PMID- 11873036 TI - Increasing maternal blood pressure with ephedrine increases uterine artery blood flow velocity during uterine contraction. AB - BACKGROUND: During labor, ephedrine is widely used to prevent or to treat maternal arterial hypotension and restore uterine perfusion pressure to avoid intrapartum fetal asphyxia. However, the effects of ephedrine on uterine blood flow have not been studied during uterine contractions. The purpose of the study was to assess the effects of ephedrine on uterine artery velocities and resistance index using the Doppler technique during the active phase of labor. METHODS: Ten normotensive, healthy parturients with uncomplicated pregnancies at term received intravenous ephedrine during labor to increase mean arterial pressure up to a maximum of 20% above their baseline pressure. Peak systolic and end-diastolic Doppler flow velocities and resistance indices were measured in the uterine artery before and immediately after administration of bolus intravenous ephedrine and after ephedrine washout. Umbilical and fetal middle cerebral arterial resistance indices and fetal heart rate were also calculated. RESULTS: After ephedrine administration, mean arterial pressure increased by 17 +/- 4%. End-diastolic flow velocity in the uterine artery at peak amplitude of uterine contraction was restored to 74% of the value observed in the absence of contraction. The systolic velocity was totally restored, and the uterine resistance index was significantly decreased, compared with the values in the absence of contraction. Between uterine contractions, ephedrine induced similar but less marked effects. Fetal hemodynamic parameters were not altered by ephedrine administration. CONCLUSIONS: Bolus administration of intravenous ephedrine reversed the dramatic decrease in diastolic uteroplacental blood flow velocity and the increase in resistance index during uterine contraction, without altering fetal hemodynamic parameters. This suggests that the increase in uterine perfusion pressure during labor could in part restore uterine blood flow to the placenta during uterine contraction. PMID- 11873037 TI - Pharmacology of spinal glutamatergic receptors in post-thermal injury-evoked tactile allodynia and thermal hyperalgesia. AB - BACKGROUND: After a focal thermal injury to the heel of a rat, thermal hyperalgesia appears at the injury site (primary thermal hyperalgesia), and tactile allodynia appears at the off-injury site (secondary tactile allodynia). The pharmacology of spinal glutamatergic receptors in the initiation and maintenance of secondary tactile allodynia was examined. METHODS: In rats prepared with chronic intrathecal catheters, the heel of one hind paw was exposed to a 52 degrees C surface for 45 s, resulting in a local erythema without blistering. Intrathecal N-methyl-d-aspartate (NMDA) receptor antagonists (MK-801, AP5) and alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid-kainate (AMPA KA) receptor antagonists (CNQX, NBQX, NS257, etc.) were administered either before (pretreatment) or after (posttreatment) the induction of the injury. Tactile withdrawal thresholds and thermal paw withdrawal latencies were assessed. RESULTS: Pretreatment and posttreatment with AMPA-KA antagonists produced a dose dependent blockade of secondary tactile allodynia. However, NMDA antagonists, in doses that effectively block other models of facilitated states, showed little or no effect. Primary thermal hyperalgesia was blocked only by high-dose AMPA-KA antagonists. CONCLUSION: Spinal AMPA-KA receptors play a major role in the initiation of secondary tactile allodynia induced by focal thermal injury. In contrast, spinal NMDA receptors play only a minimal role. PMID- 11873038 TI - Magnesium increases morphine analgesic effect in different experimental models of pain. AB - BACKGROUND: An excess of excitatory pathway activation via N-methyl-D-aspartate (NMDA) receptors has been described in neuropathic pain that responds poorly to morphine. However, in this situation, several published data sets show that coadministration of NMDA receptor antagonists restores the efficacy of opioids. Considering that magnesium behaves like an NMDA receptor antagonist, we investigated the effect of the combination of magnesium and morphine in experimental models of chronic and tonic pain. METHODS: Mechanical hyperalgesia was assessed with the paw-pressure test in mononeuropathic (chronic constrictive injury model) and diabetic rats. Behavioral reactions were scored in a model of inflammation induced by formalin. The animals were assigned to one of three groups according to the intraperitoneal pretreatment: magnesium (30 mg/kg x 3), magnesium (30 mg/kg), and saline. Before testing, morphine was injected intravenously in mononeuropathic (0.3 mg/kg) and diabetic rats (1 mg/kg) and by the subcutaneous route in rats with the formalin test (1.5 mg/kg). RESULTS: Magnesium alone induced a significant antihyperalgesic effect in mononeuropathic and diabetic rats after a cumulative dose of 90 mg/kg. Furthermore, it significantly increased morphine analgesia, regardless of the loading dose used (30 or 90 mg/kg) in the two models of neuropathic pain. In the formalin test, magnesium alone did not have a significant effect. However, in combination with morphine, it revealed the analgesic effect of this opiate. CONCLUSIONS: These data show that magnesium amplifies the analgesic effect of low-dose morphine in conditions of sustained pain. Considering the good tolerability of magnesium, these findings may have clinical applications in neuropathic and persistent pain. PMID- 11873039 TI - A combination of gabapentin and morphine mediates enhanced inhibitory effects on dorsal horn neuronal responses in a rat model of neuropathy. AB - BACKGROUND: Peripheral nerve damage can result in severe, long-lasting pain accompanied by sensory deficits. This neuropathic pain remains a clinical problem, and effective morphine analgesia is often limited by intolerable side effects. The antiepileptic gabapentin has recently emerged as an alternative chronic pain treatment. Improved management of the diverse symptoms and mechanisms of neuropathic pain may arise from combination therapy, based on multiple pharmacologic targets and low drug doses. METHODS: The authors used the Kim and Chung rodent model of neuropathy to induce mechanical and cold allodynia in the ipsilateral hind paw. In vivo electrophysiologic techniques were subsequently used to record evoked dorsal horn neuronal responses in which the effects of systemic morphine and gabapentin were investigated, both individually and in combination. RESULTS: Morphine (1 and 4 mg/kg) inhibited neuronal responses of control rats but not after neuropathy. Gabapentin (10 and 20 mg/kg) inhibited neuronal responses in nerve injured rats and to a lesser extent in sham rats but not in naive rats. In the presence of gabapentin (ineffective low dose of 10 mg/kg), morphine (1 and 3 mg/kg) mediated significant inhibitory effects in all experimental groups, with the greatest inhibitions observed in spinal nerve ligated and sham-operated rats. After neuropathy, inhibitions mediated by morphine were significantly increased in the presence of gabapentin compared with morphine alone. CONCLUSIONS: After spinal nerve ligation, the inhibitory effects of systemic morphine on evoked dorsal horn neuronal responses are reduced compared with control, whereas the effectiveness of systemic gabapentin is enhanced. In combination with low-dose gabapentin, significant improvement in the effectiveness of morphine is observed, which demonstrates a clinical potential for the use of morphine and gabapentin combinational treatment for neuropathic pain. PMID- 11873040 TI - Comparison of the effects of racemic bupivacaine, levobupivacaine, and ropivacaine on ventricular conduction, refractoriness, and wavelength: an epicardial mapping study. AB - BACKGROUND: The study was designed to compare the effects of equimolar concentrations of racemic bupivacaine, levobupivacaine, and ropivacaine on ventricular conduction, anisotropy, duration and homogeneity of refractoriness, and wavelengths, and to provide a potency ratio for effects on conduction velocity. METHODS: Isolated frozen rabbit hearts (which leave a thin layer of surviving epicardial muscle) were treated with 0.1, 1, and 10 mum racemic bupivacaine, levobupivacaine, or ropivacaine. Left ventricular longitudinal and transverse conduction velocities, anisotropic ratio, minimum pacing cycle length, use dependency, duration and dispersion of ventricular effective refractory period, and wavelengths were studied. A high-resolution mapping system was used for data acquisition. In addition to two-way analysis of variance for repeated measures, data for conduction velocities were fitted simultaneously using a nonlinear mixed-effect modeling program to allow intergroup comparison. RESULTS: Each agent induced a concentration- and use-dependent slowing of conduction velocities, with no change of the anisotropic ratio. The use-dependent effect of levobupivacaine is similar to that of racemic bupivacaine concerning longitudinal conduction velocity. Fitting of conduction velocities provided a racemic bupivacaine to levobupivacaine and to ropivacaine ratio of 1:1.38 for concentration effect at 1,000-ms pacing cycle length, and 1:0.74 for use dependent effect at 600-ms pacing cycle length. Racemic bupivacaine and levobupivacaine prolonged the ventricular effective refractory period, whereas ropivacaine did not. No dispersion in ventricular effective refractory period values occurred. All three agents induced significant decreases in wavelengths. This effect was not different among groups. CONCLUSIONS: Differences among racemic bupivacaine, levobupivacaine, and ropivacaine at equimolar concentrations are mainly caused by the use-dependent effects on conduction velocities and the concentration-dependent effects on ventricular effective refractory period. Therefore, one must take into account the corresponding pacing rates when comparing the potency ratios of local anesthetics. PMID- 11873042 TI - Ketamine inhibits sodium currents in identified cardiac parasympathetic neurons in nucleus ambiguus. AB - BACKGROUND: Ketamine increases both blood pressure and heart rate, effects commonly thought of as sympathoexcitatory. The authors investigated the possibility that ketamine increases heart rate by inhibiting the central cardiac parasympathetic mechanisms. METHODS: We used a novel in vitro approach to study the effect of ketamine on the identified cardiac parasympathetic preganglionic neurons in rat brainstem slices. The cardiac parasympathetic neurons in the nucleus ambiguus were retrogradely prelabeled with the fluorescent tracer by placing rhodamine into the pericardial sac. Dye-labeled neurons were visually identified for patch clamp recording, and ketamine effects on isolated potassium (K+) and sodium (Na+) currents were studied. RESULTS: Cardiac nucleus ambiguus neurons (n = 14) were inherently silent, but depolarization evoked sustained action potential trains with little delay or adaptation. Ketamine (10 microm) reduced this response but had no effect on the voltage threshold for action potentials (n = 14; P > 0.05). The current-voltage relations for the transient K+ current and the delayed rectified K+ current (n = 5) were unaltered by ketamine (10 mum-1 mm). Ketamine depressed the total Na+ current dose-dependently (10 microm-1 mm). In addition, ketamine shifted the Na+ current inactivation curves to more negative potentials, thus suggesting the enhancement of the Na+ channel inactivation (P < 0.05; n = 7). In the presence of Cd2+, ketamine (10 mum) continued to inhibit voltage-gated Na+ currents, which recovered completely within 10 min. CONCLUSIONS: Ketamine inhibits Na+ but not K+ channel function in brainstem parasympathetic cardiac neurons, and such actions may mediate the decrease in parasympathetic cardiac activity and increase in heart rate that occurs with ketamine. PMID- 11873041 TI - Mechanisms involved in the antiplatelet activity of midazolam in human platelets. AB - BACKGROUND: Midazolam is widely used as a sedative and anesthetic induction agent. The aim of this study was to systematically examine the inhibitory mechanisms of midazolam in platelet aggregation. METHODS: The inhibitory mechanisms of midazolam in platelet aggregation were explored by means of analysis of the platelet glycoprotein IIb-IIIa complex, phosphoinositide breakdown, intracellular Ca+2 mobilization, measurement of membrane fluidity, thromboxane B2 formation, and protein kinase C activity. RESULTS: In this study, midazolam dose-dependently (6-26 microm) inhibited platelet aggregation in human platelets stimulated by agonists. Midazolam also dose-dependently inhibited phosphoinositide breakdown and intracellular Ca+2 mobilization in human platelets stimulated by collagen. Midazolam (6-26 mum) significantly inhibited thromboxane A2 formation stimulated by collagen in human platelets. Moreover, midazolam (15 and 26 mum) dose-dependently decreased the fluorescence of platelet membranes tagged with diphenylhexatriene. Rapid phosphorylation of a platelet protein of Mr 47,000 (P47), a marker of protein kinase C activation, was triggered by collagen (2 microg/ml). This phosphorylation was markedly inhibited by midazolam (26 microm). CONCLUSIONS: These results indicate that the antiplatelet activity of midazolam may be involved in the following pathways: the effects of midazolam may initially be caused by induction of conformational changes in platelet membrane, leading to a change in the activity of phospholipase C, and subsequent inhibition of phosphoinositide breakdown and thromboxane A2 formation, thereby leading to inhibition of both intracellular Ca+2 mobilization and phosphorylation of P47 protein. PMID- 11873043 TI - Ketamine inhibits presynaptic and postsynaptic nicotinic excitation of identified cardiac parasympathetic neurons in nucleus ambiguus. AB - BACKGROUND: Ketamine increases both blood pressure and heart rate, effects commonly thought of as sympathoexcitatory. The authors investigated possible central nervous system actions of ketamine to inhibit cardiac parasympathetic neurons in the brainstem by inhibiting multiple nicotinic excitatory mechanisms. METHODS: The authors used a novel in vitro approach to study the effect of ketamine on identified cardiac parasympathetic preganglionic neurons in rat brainstem slices. The cardiac parasympathetic neurons in the nucleus ambiguus were retrogradely prelabeled with the fluorescent tracer by placing rhodamine into the pericardial sac. Dye-labeled neurons were visually identified for patch clamp recording. The effects of ketamine were tested on nicotine-evoked ligand gated currents and spontaneous glutamatergic miniature synaptic currents (mini) in cardiac parasympathetic preganglionic neurons. RESULTS: Ketamine (10 microm) inhibited (1) the nicotine (1 microm)-evoked presynaptic facilitation of glutamate release (mini frequency, 18 +/- 7% of control; n = 9), and (2) the direct postsynaptic ligand-gated current (27 +/- 8% of control; n = 9), but ketamine did not alter the amplitude of postsynaptic miniature non-N-methyl-D aspartate currents. alpha Bungarotoxin, an antagonist of alpha 7 containing nicotinic presynaptic receptors, blocked ketamine actions on mini frequency (n = 10) but not mini amplitude. CONCLUSIONS: Ketamine inhibits the presynaptic nicotinic receptors responsible for facilitating neurotransmitter release, as well as the direct ligand-gated inward current, but does not alter the nicotinic augmentation of non-N-methyl-D-aspartate currents in brainstem parasympathetic cardiac neurons. Such actions may mediate the decrease in parasympathetic cardiac activity and increase in heart rate that occurs with ketamine. PMID- 11873044 TI - Is isoflurane-induced preconditioning dose related? AB - BACKGROUND: Volatile anesthetics precondition against myocardial infarction, but it is unknown whether this beneficial action is threshold- or dose-dependent. The authors tested the hypothesis that isoflurane decreases myocardial infarct size in a dose-dependent fashion in vivo. METHODS: Barbiturate-anesthetized dogs (n = 40) were instrumented for measurement of systemic hemodynamics including aortic and left ventricular pressures and rate of increase of left ventricular pressure. Dogs were subjected to a 60-min left anterior descending coronary artery occlusion followed by 3 h of reperfusion and were randomly assigned to receive either 0.0, 0.25, 0.5, 1.0, or 1.25 minimum alveolar concentration (MAC) isoflurane in separate groups. Isoflurane was administered for 30 min and discontinued 30 min before left anterior descending coronary artery occlusion. RESULTS: Infarct size (triphenyltetrazolium staining) was 29 +/- 2% of the area at risk in control experiments (0.0 MAC). Isoflurane produced significant (P < 0.05) reductions of infarct size (17 +/- 3, 13 +/- 1, 14 +/- 2, and 11 +/- 1% of the area at risk during 0.25, 0.5, 1.0, and 1.25 MAC, respectively). Infarct size was inversely related to coronary collateral blood flow (radioactive microspheres) in control experiments and during low (0.25 or 0.5 MAC) but not higher concentrations of isoflurane. Isoflurane shifted the linear regression relation between infarct size and collateral perfusion downward (indicating cardioprotection) in a dose-dependent fashion. CONCLUSIONS: Concentrations of isoflurane as low as 0.25 MAC are sufficient to precondition myocardium against infarction. High concentrations of isoflurane may have greater efficacy to protect myocardium during conditions of low coronary collateral blood flow. PMID- 11873045 TI - Extracellular magnesium ion modifies the actions of volatile anesthetics in area CA1 of rat hippocampus in vitro. AB - BACKGROUND: Magnesium ion (Mg2+) is involved in important processes as modulation of ion channels, receptors, neurotransmitter release, and cell excitability in the central nervous system. Although extracellular Mg2+ concentration ([Mg2+]o) can be altered during general anesthesia, there has been no evidence for [Mg2+]o dependent modification of anesthetic actions on neural excitability in central nervous system preparations. The purpose of current study was to determine whether the effects of volatile anesthetics are [Mg2+]o-dependent in mammalian central nervous system. METHODS: Extracellular electrophysiologic recordings from CA1 neurons in rat hippocampal slices were used to investigate the effects of [Mg2+]o and anesthetics on population spike amplitude and excitatory postsynaptic potential slope. RESULTS: The depression of population spike amplitudes and excitatory postsynaptic potential slopes by volatile anesthetics were significantly dependent on [Mg2+]o. The effects were attenuated in the presence of a constant [Mg2+]o/extracellular Ca2+ concentration ratio. However, neither N methyl-d-aspartate receptor antagonists nor a non-N-methyl-d-aspartate receptor antagonist altered the [Mg2+]o-dependent anesthetic-induced depression of population spikes. Volatile anesthetics produced minimal effects on input-output (excitatory postsynaptic potential-population spike) relations or the threshold for population spike generation. The effects were not modified by changes in [Mg2+]o. In addition, the population spike amplitudes, elicited via antidromic (nonsynaptic) stimulation, were not influenced by [Mg2+]o in the presence of volatile anesthetics. CONCLUSIONS: These results provide support that alteration of [Mg2+]o modifies the actions of volatile anesthetics on synaptic transmission and that the effects could be, at least in part, a result of presynaptic Ca2+ channel-related mechanisms. PMID- 11873046 TI - Propofol attenuates beta-adrenoreceptor-mediated signal transduction via a protein kinase C-dependent pathway in cardiomyocytes. AB - BACKGROUND: Activation of beta adrenoreceptors by catecholamines is an important mechanism for increasing the inotropic state of the heart. The objectives of the current study were to investigate the effects of propofol on beta-adrenoreceptor mediated increases in cardiomyocyte intracellular Ca2+ concentration ([Ca2+]i), cell shortening, L-type Ca2+ current (ICa) and cyclic adenosine monophosphate (cAMP) accumulation. The authors also investigated the site of action of propofol in the beta-adrenoreceptor signaling pathway, as well as the role of protein kinase C (PKC), and tested the hypothesis that propofol would inhibit the beta adrenoreceptor signaling pathway via a PKC-dependent mechanism. METHODS: Freshly isolated ventricular myocytes were obtained from adult rat and guinea pig hearts. Myocyte shortening (video edge detection) and [Ca2+]i (fura-2, 340/380 ratio) were monitored simultaneously in individual cells. Conventional whole cell patch clamp analysis was used to measure the ICa in individual myocytes. cAMP production was assessed in suspensions of myocytes using an enzyme immunoassay kit. RESULTS: Propofol (0.1-10 mum) had no effect on steady state [Ca2+]i, cell shortening, ICa, or cAMP production. In contrast, propofol caused dose-dependent decreases in isoproterenol-stimulated increases in [Ca2+]i, shortening, ICa, and cAMP. Forskolin-induced increases in [Ca2+]i, shortening, and cAMP production were not altered by propofol. PKC activation with phorbol myristate acetate attenuated isoproterenol-stimulated cAMP production. Inhibition of PKC with bisindolylmaleimide (broad range inhibitor) or Go 6976 (inhibitor of Ca2+ dependent PKC isoforms) abolished propofol-induced inhibition of isoproterenol stimulated increases in [Ca2+]i, shortening, and cAMP production. CONCLUSIONS: Clinically relevant concentrations of propofol attenuate beta-adrenergic signal transduction in cardiac myocytes via inhibition of cAMP production. The inhibitory site of action of propofol is upstream of adenylyl cyclase and involves activation of PKC alpha. PMID- 11873047 TI - The diverse actions of volatile and gaseous anesthetics on human-cloned 5 hydroxytryptamine3 receptors expressed in Xenopus oocytes. AB - BACKGROUND: General anesthetics can modulate the 5-hydroxytryptamine type 3 (5 HT3) receptor, which may be involved in processes mediating nausea and vomiting, and peripheral nociception. The effects of the new volatile anesthetic sevoflurane and the gaseous anesthetics nitrous oxide (N2O) and xenon (Xe) on the 5-HT3 receptor have not been well-characterized. METHODS: Homomeric human-cloned 5-HT3A receptors were expressed in Xenopus oocytes. The effects of halothane, isoflurane, sevoflurane, N2O, and Xe on 5-HT-induced currents were studied using a two-electrode, voltage clamping technique. RESULTS: Halothane (1%) and isoflurane (1%) potentiated 1 mum 5-HT-induced currents to 182 +/- 12 and 117 +/- 2%, respectively. In contrast, sevoflurane (1%), N2O (70%), and Xe (70%) inhibited 5-HT-induced currents to 76 +/- 1, 77 +/- 4, and 34 +/- 4%, respectively. The inhibitory effects were noncompetitive for sevoflurane and competitive for N2O and Xe. None of these inhibitory effects showed voltage dependency. CONCLUSION: Inhalational general anesthetics produce diverse effects on the 5-HT3 receptor. Both halothane and isoflurane enhanced 5-HT3 receptor function in a concentration-dependent manner, which is consistent with previous studies. Sevoflurane inhibited the 5-HT3 receptor noncompetitively, whereas N2O and Xe inhibited the 5-HT3 receptor competitively, suggesting the inhibitory mechanism of sevoflurane might be different from those of N2O and Xe. PMID- 11873048 TI - Effects of dantrolene on extracellular glutamate concentration and neuronal death in the rat hippocampal CA1 region subjected to transient ischemia. AB - BACKGROUND: Excessive extracellular glutamate produced by cerebral ischemia has been proposed to initiate the cascade toward neuronal cell death. Changes in extracellular glutamate concentration are closely linked to changes in intracellular calcium ion concentration. Dantrolene inhibits calcium release from intracellular calcium stores. In this study, the authors investigated the effects of dantrolene on extracellular glutamate accumulation and neuronal degeneration in a rat model of transient global forebrain ischemia. METHODS: Male Wistar rats weighing 230-290 g were anesthetized with halothane in nitrous oxide-oxygen and were subjected to 10 min of transient forebrain ischemia using a four-vessel occlusion technique. Fifteen minutes before ischemic injury, dantrolene sodium (5 mm), dimethyl sulfoxide as a vehicle for dantrolene, or artificial cerebrospinal fluid as a control was intracerebroventricularly administered (n = 8 in each group). In the hippocampal CA1 subfield, the extracellular glutamate concentration in vivo was measured during the periischemic period with a microdialysis biosensor, and the number of intact neurons was evaluated on day 7 after reperfusion. RESULTS: Both dantrolene and dimethyl sulfoxide significantly reduced the ischemia-induced increase in glutamate concentration to a similar extent, i.e., by 53 and 51%, respectively, compared with artificial cerebrospinal fluid (P < 0.01). The number of intact hippocampal CA1 neurons (mean +/- SD; cells/mm) in dantrolene-treated rats (78 +/- 21) was significantly higher than that in artificial cerebrospinal fluid- (35 +/- 14; P < 0.001) and dimethyl sulfoxide-treated (56 +/- 11; P < 0.05) animals. Dimethyl sulfoxide also significantly increased the number of preserved neurons in comparison with artificial cerebrospinal fluid (P < 0.05). CONCLUSIONS: Intracerebroventricular dantrolene prevents delayed neuronal loss in the rat hippocampal CA1 region subjected to transient ischemia; however, this neuroprotection cannot be accounted for only by the reduced concentrations of extracellular glutamate during ischemia. PMID- 11873049 TI - Pharmacodynamic modeling of muscle relaxants: effect of design issues on results. AB - BACKGROUND: Pharmacodynamic studies of muscle relaxants use different dosing regimens (such as administration by bolus vs. infusion and doses that produce complete vs. incomplete paralysis). The authors used published data to evaluate the effect of modeling assumptions on pharmacodynamic estimates. METHODS: The authors used a pharmacokinetic-pharmacodynamic dataset in which patients received cisatracurium, 75 or 300 microg/kg (1.5 or 6 x ED95), to generate plasma concentration (Cp) and twitch depression (effect) curves. They then evaluated the impact of the following: assuming that Cp decreased monotonically versus increasing initially before decreasing monotonically; misrecording effect data by 6 s or less; and doses targeting incomplete versus complete paralysis. Parameters evaluated were the steady state Cp depressing twitch tension 50% (C50) and the rate constant for equilibration between plasma and effect site concentrations (k(e0)). RESULTS: With the large dose, increasing the time at which Cp peaked from 0.0 to 1.5 min decreased C50 and increased k(e0) markedly; with the small dose, changes in both were small. Misrecording the timing of effect had a larger impact with the large dose compared with the small dose. Doses smaller than ED50 or those producing prolonged, complete twitch depression yielded biased and variable estimates. CONCLUSION: The erroneous assumption that Cp decreases monotonically after bolus administration affects accuracy of pharmacodynamic estimates with doses producing rapid, complete twitch depression. Other errors (e.g., misrecording the time of drug administration) impact on pharmacodynamic estimates, particularly with large doses. The authors' findings suggest that investigators performing neuromuscular (and other) pharmacodynamic studies should carefully consider the impact of study design on their parameter estimates. PMID- 11873050 TI - Use of linear programming to estimate impact of changes in a hospital's operating room time allocation on perioperative variable costs. AB - BACKGROUND: Administrators at hospitals with a fixed annual budget may want to focus surgical services on priority areas to ensure its community receives the best health services possible. However, many hospitals lack the detailed managerial accounting data needed to ensure that such a change does not increase operating costs. The authors used a detailed hospital cost database to investigate by how much a change in allocations of operating room (OR) time among surgeons can increase perioperative variable costs. METHODS: The authors obtained financial data for all patients who underwent outpatient or same-day admit surgery during a year. Linear programming was used to determine by how much changing the mix of surgeons can increase total variable costs while maintaining the same total hours of OR time for elective cases. RESULTS: Changing OR allocations among surgeons without changing total OR hours allocated will likely increase perioperative variable costs by less than 34%. If, in addition, intensive care unit hours for elective surgical cases are not increased, hospital ward occupancy is capped, and implant use is tracked and capped, perioperative costs will likely increase by less than 10%. These four variables predict 97% of the variance in total variable costs. CONCLUSIONS: The authors showed that changing OR allocations among surgeons without changing total OR hours allocated can increase hospital perioperative variable costs by up to approximately one third. Thus, at hospitals with fixed or nearly fixed annual budgets, allocating OR time based on an OR-based statistic such as utilization can adversely affect the hospital financially. The OR manager can reduce the potential increase in costs by considering not just OR time, but also the resulting use of hospital beds and implants. PMID- 11873051 TI - A qualitative and quantitative systematic review of preemptive analgesia for postoperative pain relief: the role of timing of analgesia. PMID- 11873052 TI - Practice guidelines for postanesthetic care: a report by the American Society of Anesthesiologists Task Force on Postanesthetic Care. PMID- 11873053 TI - Lethal air embolism during cesarean delivery for placenta previa. PMID- 11873054 TI - High gastric output as a perioperative sign of carcinoid syndrome. PMID- 11873055 TI - Subcellular localization of trifluoroacetylated liver proteins in association with hepatitis following isoflurane. PMID- 11873056 TI - Bilateral continuous interscalene block of brachial plexus for analgesia after bilateral shoulder arthroplasty. PMID- 11873057 TI - Delayed postoperative rhabdomyolysis in a patient subsequently diagnosed as malignant hyperthermia susceptible. PMID- 11873058 TI - Ventilatory failures with the Datex-Ohmeda 7900 SmartVent. PMID- 11873059 TI - Clinical and experimental research in anesthesiology in Europe at the change of the millennium. PMID- 11873060 TI - Clinical and experimental research in anesthesiology in Europe at the change of the millennium. PMID- 11873061 TI - Propofol, metabisulfite, and bronchoconstriction. PMID- 11873062 TI - Electrocautery-induced pacemaker tachycardia: why does this error continue? PMID- 11873063 TI - The effect of edrophonium on autonomic outflow. PMID- 11873064 TI - World War II and physician specialization. PMID- 11873065 TI - Calling all anesthetists to service in World War II. PMID- 11873066 TI - The LTA cannula and difficult intubations. PMID- 11873067 TI - Another technique of facilitating a difficult intubation. PMID- 11873068 TI - Subtle hyperkalemia detected through monitor "artifact". PMID- 11873070 TI - Fetal and maternal outcome after administration of tenofovir to gravid rhesus monkeys (Macaca mulatta). AB - Tenofovir has been shown to cross the placenta in quantities sufficient to sustain reductions in viral load in simian immunodeficiency virus (SIV)-infected fetal monkeys. With chronic exposure (30 mg/kg), however, significant bone related toxicity has been shown in approximately 25% of infants studied. Further investigations were conducted to determine whether the bone-related toxicity observed was initiated during fetal life. Gravid rhesus monkeys (n = 4) were administered tenofovir subcutaneously once daily from 20 to 150 days of gestation (30 mg/kg; term: 165 +/- 10 days). Fetuses were monitored sonographically, and maternal and fetal blood and urine samples were collected to assess hematologic parameters, clinical chemistry, insulin-like growth factor (IGF) levels, and bone biomarkers. Fetuses were delivered by hysterotomy near term for necropsy and evaluation of bone-related mechanical properties. Results of these studies have shown 1) normal fetal development, although overall body weights and crown-rump lengths were less than those for age-matched controls (p < or = .03); 2) a significant reduction in circulating IGF-I (p <.001); 3) a small reduction in fetal bone porosity (p < or = .03); and 4) transient alterations in maternal body weights and bone-related biomarkers during the treatment period. The results of these studies suggest that chronic fetal exposure to tenofovir at the maternal dose of 30 mg/kg throughout gestation can alter select fetal parameters and transiently affect maternal bone biomarkers. PMID- 11873071 TI - Randomized, open-label study of the impact of two doses of subcutaneous recombinant interleukin-2 on viral burden in patients with HIV-1 infection and CD4+ cell counts of > or = 300/mm3: CPCRA 059. AB - The effect of intermittent courses of recombinant interleukin-2 (rIL-2) on HIV-1 load in patients receiving combination antiretroviral therapy remains uncertain. CPCRA 059 was an open-label, randomized, multicenter trial in which 511 patients with HIV-1 infection and CD4+ cell counts of > or = 300/mm3 who were receiving antiretroviral therapy were assigned to receive no rIL-2 (255 patients [controls]) or subcutaneous rIL-2 in dosages of 4.5 MIU (130) or 7.5 MIU (126) twice daily for 5-day courses every 8 weeks to maintain CD4+ cell counts that were twice the baseline value or > or = 1,000/mm3. The primary objective of this study was to compare the effects of the two doses of rIL-2 and no rIL-2 on viral load and CD4+ cell counts over 12 months. There was no difference in the following viral load measurements between the rIL-2 treatment groups and the control treatment group: percentage of patients with viral loads of <50 copies/mL at 12 months (p =.55), time to viral load of > or = 50 copies/mL for patients who had baseline viral loads of <50 copies/mL (p =.35), and change in viral load from baseline for patients who had viral loads of > or = 50 copies/mL at baseline (p =.63). At each follow-up visit, the change in CD4+ cell count from baseline was significantly greater in the rIL-2 treatment groups than in the control treatment group, with a mean difference of 251/mm3 at month 12 (95% confidence interval, 207-295; p <.0001). No unanticipated adverse experiences were seen in this trial, to our knowledge the largest randomized evaluation of rIL-2 treatment conducted to date. PMID- 11873073 TI - Quality of life, emotional status, and adherence of HIV-1-infected patients treated with efavirenz versus protease inhibitor-containing regimens. AB - We assessed the impact of an efavirenz-containing regimen versus a protease inhibitor-containing regimen on quality of life, emotional status, and adherence of HIV-1-infected patients. In addition, we sought to define the adverse events associated with these treatments, with a special focus on central nervous system disorders in the efavirenz treatment group. This prospective, randomized, two arm, controlled study included 100 patients for whom initial treatment with a protease inhibitor-containing regimen failed. Patients were randomized to start treatment with two nucleoside retrotranscriptase inhibitors plus efavirenz (group 1; 51 patients) or two nucleoside retrotranscriptase inhibitors plus one or more new protease inhibitors (group 2; 49 patients). Quality of life was assessed by a five-point item adapted from the HIV questionnaire of the Medical Outcomes Study, emotional status was evaluated by the Profile of Mood State questionnaire, and patients self-reported adherence. Data were analyzed by both an as-treated method and an intention-to-treat-last observation carried forward method. Patients in group 1 reported the following findings at week 4: dizziness (66%), abnormal dreaming (48%), light-headedness (37%), and difficulty sleeping (35%). At week 24, dizziness (13%; p <.001), abnormal dreaming (18%; p =.002), light-headedness (13%; p =.01), difficulty sleeping (7%; p =.001), and nervousness (13%; p =.01) decreased in these patients. Irritability, abnormal dreaming, and nervousness persisted at week 48 in 13%, 10%, and 8% of group 1 patients, respectively. Patients in group 2 reported the following findings at week 4: light-headedness (8%), dizziness (5%), difficulty sleeping (4%), nervousness (4%), and headaches (3%). Patients in group 2 reported the following findings at week 48: difficulty sleeping (4%), nervousness (3%), headaches (3%), and light-headedness (2%). In group 1, quality of life (p <.001) and emotional status (week 48; p =.004) improved, both of which were better than those in group 2 (p =.001). Both groups maintained high levels of medication adherence, and no significant differences in the number of patients who had viral loads of <200 copies/mL at week 48 were found (78% of group 1 patients vs. 85% of group 2 patients; p = not significant). At week 48, the mean CD4 cell count +/- SD was 497 +/- 224/mm3 in group 1 and 539 +/- 298/mm3 in group 2 (p = not significant). Despite similar immunologic and virologic outcomes, a second-line efavirenz-containing regimen improved quality of life of HIV-1-infected patients compared with a second-line protease inhibitor containing regimen. However, close follow-up of patients receiving treatment with efavirenz-based regimens is recommended, especially for those with previous emotional disturbances due to central nervous system disorders in the short term and those with persistence of a low percentage of these disorders in the long term. PMID- 11873072 TI - Concordance between HIV source partner identification and molecular confirmation in acute retroviral syndrome. AB - Most HIV-1 transmission studies use self-reported history to define the source contact. To evaluate the reliability of epidemiologic source partner reporting, heteroduplex mobility assays (HMAs) were performed comparing the different viral strains present in the partners. Partners were typed for human leukocyte antigen (HLA) to evaluate the degree of shared alleles. Of 11 couples evaluated, HMA analysis confirmed nine transmissions (including 1 oral-genital transmission), indicated probable transmission in 1 couple, and suggested an alternative source partner in another. Nine source partners transmitted a major variant. Four source partners knew their HIV status. Previous HIV monitoring was reported by 5 of the 6 confirmed source partners who were unaware of their HIV status at the time of transmission. We also evaluated potential "sharing of HLA alleles" as a risk factor for HIV-1 acquisition; partners were not found to have a higher degree of shared HLA alleles. Lack of awareness about infection status as a consequence of infrequent testing plays a major role in the secondary transmission of HIV. These findings re-emphasize the importance of using safe sex practices at all times. PMID- 11873074 TI - Safety and immunogenicity of a high-titered canarypox vaccine in combination with rgp120 in a diverse population of HIV-1-uninfected adults: AIDS Vaccine Evaluation Group Protocol 022A. AB - To test the safety and immunogenicity of a high-titered preparation of ALVAC-HIV vCP205 in both high-risk and low-risk persons and to evaluate variations in dosing schedule, we conducted a multicenter, randomized, double-blind trial of this vector in combination with recombinant subunit gp120 in 150 HIV-1 seronegative volunteers. The high-titered ALVAC vaccine was well tolerated; adverse events were minimal and not influenced by dosing. At day 728, the cumulative probability of a cytotoxic T-lymphocyte (CTL) response was 76% (95% confidence interval [CI]: 64%-89%) among volunteers receiving vaccine, and the net amount attributable to vaccination was 50% (CI: 16%; 74%). The net probability of a repeated positive CTL response by day 728 was 50% (CI: 21%; 64%). There was a significant difference in CTL response at day 182 between volunteers who had received four doses versus three doses of vCP205 (42% vs. 24%, p =.052). The CTL response was similar in high-risk volunteers and vaccinia-naive volunteers compared with vaccinia-immune volunteers. Neutralizing antibody responses were detected in 95% of vaccinees at day 287, with higher geometric mean titers in recipients of sequential versus simultaneous dosing of the two vaccines and in vaccinia-naive volunteers. This high-titered preparation of ALVAC HIV vCP205 in combination with gp120 was safe and immunogenic in a diverse group of HIV-1-seronegative volunteers. PMID- 11873075 TI - Placental inflammation and perinatal transmission of HIV-1. AB - The effect of placental membrane inflammation on mother-to-child transmission (MTCT) of HIV-1 is reported. Placentas from HIV-1-infected women were examined as part of a perinatal HIV-1 project in Mombasa, Kenya. Polymerase chain reaction analysis was used to test for HIV-1 in the infants at birth and at 6 weeks. The maternal HIV-1 seroprevalence was 13.3% (298 of 2,235). The overall rate of MTCT of HIV-1 was 25.4%; polymerase chain reaction analysis revealed that of the 201 infants 6.0% (12) were already HIV-1-positive at birth (intrauterine transmission) and 19.4% (39) were infected during the peripartum period or in early neonatal life (perinatal transmission). The prevalence of acute chorioamnionitis was 8.8%, that of deciduitis was 10.8%, and that of villitis was 1.6%. Acute chorioamnionitis was independently associated with peripartum HIV-1 transmission but not with in utero MTCT (17.9% vs. 6.7%, respectively; adjusted odds ratio, 3.9; 95% confidence interval, 1.2-12.5; p =.025). Other correlates of perinatal MTCT were presence of HIV in the genital tract and in the baby's oral cavity and a high maternal viral load in peripheral blood. The adjusted population attributable fraction of 12.8% (95% confidence interval, 1.5%-22.8%) indicated that approximately 3% of MTCT could be prevented if acute chorioamnionitis was eliminated. We suggest that further research on the role of antimicrobial treatment in the prevention of chorioamnionitis and the reduction of peripartum MTCT needs to be performed. PMID- 11873076 TI - HIV-1 RNA viral load monitoring in HIV-infected drug users on antiretroviral therapy: relationship with outpatient care patterns. AB - HIV-1 viral load (VL) testing is a standard component of HIV care. We examined the use and predictors of VL testing in drug users, a group at risk for problematic care. Using 1996 to 1998 New York State (NYS) Medicaid files, we studied drug users who had been enrolled >10 months, had been prescribed antiretroviral agents in 1997 and 1998, and who had undergone any VL testing in 1997. Our outcome was regular VL testing shown by two or more paid claims for this test in 1998. Patterns of care in 1997 were defined as: regular source of medical care (>35% of visits to one provider), and/or regular drug treatment of >6 months, or neither. We counted visits in 1997 to a provider offering HIV focused care. Adjusted odds ratios (AORs) of VL testing were assessed. Of 3131 drug users, 73.9% had at least one VL test, whereas 56.2% had two or more VL tests in 1998. The AORs of two or more VL tests were increased for those with regular drug abuse care alone (AOR, 1.50; 95% confidence interval [CI], 1.21 1.84) or with regular medical care (AOR, 1.27; 95% CI, 1.03-1.57) versus those with neither. HIV-focused care was positively associated with two or more VL tests (AOR, 1.38; 95% CI, 1.05-1.81 for 1-3 visits; AOR, 1.94; 95% CI, 1.50-2.51 for four or more visits). We found that nearly half this cohort of drug users did not have regular VL testing. Drug users with HIV-focused care or with regular drug treatment are more likely to have regular VL testing. PMID- 11873078 TI - Incidence of HIV-1 infection and effects of clinic-based counseling on HIV preventive behaviors among married women in northern Thailand. AB - To determine the incidence of and risk factors for HIV-1 infection among married women in northern Thailand, we enrolled 779 seronegative women from family planning clinics and a postpartum ward in Chiang Rai, Thailand, from 1998 through 1999. Women were tested for HIV antibodies at 6 and 12 months after enrollment. They received HIV prevention counseling at enrollment and at each follow-up visit. Counseling covered partner communication, partner HIV testing, and condom use by steady partners. Effects of counseling were measured using standardized questionnaires. Follow-up rates were 94% at 6 months and 92% at 12 months. Only 1 woman seroconverted during the follow-up period, yielding an overall HIV incidence of 0.14 per 100 person-years. After receiving counseling, women reported significantly increased communication with husbands concerning HIV risk, HIV testing, and condom use during the first 6 months after enrollment; communication remained high for 6 to 12 months. Women reported a modest increase in HIV testing and consistent condom use by husbands. The risk for HIV transmission to women in steady relationships is low in northern Thailand. Although HIV prevention counseling promoted partner communication, its effects on HIV preventive behaviors were limited. PMID- 11873077 TI - Male viral load and heterosexual transmission of HIV-1 subtype E in northern Thailand. AB - We evaluated the association between HIV-1 RNA copies/mL in men and heterosexual transmission to their female partners among 493 couples in Thailand. Husbands were identified as HIV-positive when they were screened as blood donors; nearly all were infected with HIV subtype E. Wives had no known risks for HIV infection other than sex with their husbands. In multivariate analysis, each log10 increment of HIV RNA in the man was associated with an 81% increased rate of HIV transmission to his wife (odds ratio = 1.81, 95% confidence interval: 1.33-2.48). No transmission occurred at viral loads below 1094 copies/mL, and a dose-response effect was seen with increasing viral load in the man. In multivariate analysis, a history of a sexually transmitted disease in the man or woman, longer duration of hormonal contraceptive use, and the woman's onset of sexual activity at less than 20 years of age were also associated with increased seropositivity of the wife. PMID- 11873079 TI - Use of a public sexually transmitted disease clinic by known HIV-positive adults: decreased self-reported risk behavior and increased disease incidence. AB - High-risk sexual behavior by HIV-positive individuals is an important factor contributing to the spread of the HIV epidemic. We conducted a retrospective chart review to compare self-reported sexually transmitted disease (STD) risk behavior and clinic diagnoses of known HIV-positive clients attending Miami-Dade County STD clinics with those of uninfected controls. One hundred ninety-one HIV positive clients and 191 HIV-negative controls, 130 (68.1%) men and 61 (31.9%) women, were included in the analysis. HIV-positive clients were more likely than controls to report no sexual activity in the last 2 months (odds ratio [OR] = 2.6, 95% confidence interval [CI]: 1.5-4.5) or, if active, to report condom use at last sexual intercourse (OR = 3.1, CI: 1.9-5.3). However, HIV-positive clients were more likely to be diagnosed with infectious syphilis (OR = 13.0, CI: 1.6 99.4) and/or gonorrhea (OR = 2.1, CI: 1.1-4.2) than controls. This may be a result of overreporting of condom use or sexual activity in high-risk sexual networks with inefficient use of condoms. Ongoing sexual risk behavior and access to HIV primary care are important issues in this population. PMID- 11873081 TI - Detection of HTLV-I tax-rex and pol gene sequences of thymus gland in a large group of patients with myasthenia gravis. AB - We report in this study the use of polymerase chain reaction (PCR) for the amplification of the genomic DNA, isolated from thymic tissue, using the primers flanking HTLV-I/II tax-rex genes and the sequence method to analyze the HTLV-I pol sequence of 27 Italian patients with myasthenia gravis. These molecular methods showed that 92.5% of patients tested positive for tax gene and 55% for pol genes; 55.5% samples were positive for both the tax gene of HTLV-I/II, and the pol gene of HTLV-I. Histologic investigation of the thymus showed that 15 samples had thymic hyperplasia, 93% tested positive for the tax gene, and 40% tested positive for both the tax and pol genes of HTLV-I. In contrast, 91.6% of thymoma-positive samples were positive for tax gene I/II and 75% positive for both genes, tax and pol type I. The sequence analysis of PCR product for tax and pol genes confirmed that these amplified products were HTLV-I, with minimal variations. Our date suggested that either HTLV-I or part of the virus genome is involved in the etiopathogenesis of myasthenia gravis. PMID- 11873080 TI - Prevalence trend and correlates of HHV-8 infection in HIV-infected patients. AB - To assess the circulation of human herpesvirus (HHV)-8 infection over the years, two seroprevalence surveys were conducted, which tested sera from HIV-infected individuals recruited 10 years apart (206 individuals from 1986 to 1988 and 177 individuals from 1997 to 1998). For all patients, antibodies to hepatitis C virus (HCV), hepatitis B virus (HBV), and HHV-8 lytic and latent antigens were evaluated.HHV-8 seroprevalence was higher among individuals recruited in the 1990s (31.6% for anti-lytic, 8.5% for anti-latent antibodies) compared with similar findings in those seen in the late 1980s (14.6% and 3.4% for anti-lytic and anti-latent antibodies, respectively), with a twofold increase of the risk of HHV-8 infection. However, the increase was observed only among injecting drug users, whereas seroprevalence tended to slightly increase among those infected by sexual contact. At univariate analysis, time of recruitment and being homosexual men were factors associated with HHV-8 infection, an association that remained after adjusting for age. HBV infection was significantly associated with HHV-8 infection (odds ratio [OR], 2.2; 95% confidence interval [CI], 1.3-3.6), whereas those infected with HCV had a lower probability of having HHV-8 antibodies (OR, 0.3; 95% CI, 0.20-0.6). After controlling for age and gender, time of recruitment remained independently associated with HHV-8 infection among injecting drug users. In conclusion, HHV-8 seroprevalence appears to be increased during 10 years among HIV-infected injection drug users but not among homosexual men, who remain those at the highest risk of infection. PMID- 11873083 TI - Genetic analysis of HIV-1 discordant couples in Thailand: association of CCR2 64I homozygosity with HIV-1-negative status. PMID- 11873082 TI - HIV-1 infection in individuals with the CCR5-Delta32/Delta32 genotype: acquisition of syncytium-inducing virus at seroconversion. AB - Homozygosity for the 32 base-pair deletion (Delta32/Delta32) in the CCR5 coreceptor gene is associated with incomplete HIV-1 resistance. Six HIV-1 infected Delta32/Delta32 patients have been reported. We report 2 additional Delta32/Delta32-infected individuals, among 106 seroconverters in a vaccine preparedness study. Like the previous 6, these individuals experienced rapid CD4 decline. However, taken together, the 8 patients have neither uniformly high virus load nor rapid progression to AIDS. We obtained five virus isolates from 1 patient at 5, 6, 7, 10, and 12 months after the estimated time of infection. The earliest isolate exhibits the syncytium-inducing (SI) phenotype and exclusive use of the CXCR4 coreceptor, suggesting acquisition of HIV-1 through this coreceptor. Of the remaining 104 seroconverters, 8 were CCR5-Delta32/+ and 96 were CCR5-+/+. Three CCR5-+/+ seroconverters who showed the uncommon pattern of early SI virus and rapid CD4 decline had uniformly high viral load and more heterogeneous coreceptor usage. These results further support the conclusion that Delta32 mediated resistance is incomplete and is associated with acquisition of exclusively-X4 variants of HIV-1. The pathogenic potential of these viruses may be different from late-stage X4 virus or early X4 virus acquired by individuals with other CCR5 genotypes. PMID- 11873085 TI - Genotypic HIV-1 drug resistance testing in antiretroviral-naive subjects in Houston, Texas. PMID- 11873084 TI - Influence of hepatitis C virus coinfection on lipid abnormalities in HIV-positive patients after highly active antiretroviral therapy. PMID- 11873086 TI - Factors associated with the time elapsed between initial detection of HIV antibodies and first contact for healthcare in HIV seroconverters of the Lyon University Hospitals. PMID- 11873087 TI - Evaluation of two rapid HIV screening tests for the detection of HIV-2 antibody. PMID- 11873088 TI - Risk stratification of coronary heart disease through established and emerging lifestyle factors in a Mediterranean population: CARDIO2000 epidemiological study. AB - BACKGROUND: In recent decades various lifestyle factors have been associated with the increasing risk of coronary heart disease. The aim of this study is to assess coronary risk, based on established and emerging lifestyle risk factors such as smoking habit, physical activity, alcohol consumption and depression in a Mediterranean population. METHODS: During 2000--2001, 535 male and 126 female patients with a first event of an acute coronary syndrome and 661 controls matched by sex, age and region were entered into the study. Conditional logistic regression analysis estimated the relative risks of developing coronary heart disease under several scenarios. RESULTS: The multivariate analysis showed that stopping smoking is exponentially related to the reduction of coronary risk. By contrast, passive smoking increases the risk from 48% to 112%. Familial smoking habits are related to current smoking status, increasing the coronary risk fourfold. The effect of depression on coronary risk differs according to gender (+15% in males vs. 32% in females), and interacts with retirement, current smoking, physical inactivity, alcohol consumption and social status, increasing the risk from 15% to 189%. A J-shape association was found between alcohol intake and coronary risk Physical inactivity doubles the coronary risk, while long-term physical activity plays a preventive role in the reduction of coronary risk, even in the elderly. CONCLUSION: Analysis of the data showed that the cluster of established and emerging lifestyle factors poses its particular role for further investigation in the aetiology of coronary heart disease. PMID- 11873089 TI - Group- and sex-specific effects of age, body composition and pulse rate on blood pressure variability in some cross-cultural populations of Visakhapatnam District; South India. AB - BACKGROUND: Hypertension is becoming a major public health problem in developing countries. Blood pressure tends to vary cross-culturally and is influenced by various factors including age, body composition and pulse rate. This paper reports the influence of age, body composition and pulse rate on blood pressure variability. DESIGN: The data were collected from 1316 individuals (646 men and 670 women) belonging to two tribes (Khondh and Valmiki) inhabiting hilly tribal area and two castes (Wadabalija and Settibalija) from both rural and urban areas from Visakhapatnam district, Andhra Pradesh, India. RESULTS: There are considerable ethnic differences in blood pressure and body composition variables. Age contributes significantly to the variance in blood pressure in these populations. However the influence of age is less pronounced in Khondh, a primitive/unacculturized tribe, while age exerts a profound effect on blood pressure variability in Valmiki, an acculturizing tribe. Further, the effect of age is more pronounced on systolic than on diastolic pressure and it contributes more to the blood pressure of women than men. The subcutaneous fatness measures (sum of three trunk skin fold thicknesses and sum of three extremities skin fold thicknesses) are significant predictors of both systolic and diastolic blood pressure in Khondh, Valmiki and Wadabalija, while body mass index (BMI) is an important predictor of both systolic and diastolic blood pressure in Settibalija. Pulse rate is often found to influence blood pressure of women. CONCLUSION: The age-associated variability of blood pressure confirms the proposition that variability is high in adverse environment, when modernization/acculturization is taken to represent environmental adversity. Apart from the influence of age, body composition/adiposity contributes significantly to the blood pressure variability. PMID- 11873090 TI - Anticardiolipin antibodies and risk of thromboembolic disease in young Jamaican women. AB - BACKGROUND: Anticardiolipin antibodies (aCL) are a heterogeneous group of antiphospholipid antibodies that are associated with arterial and venous thrombosis. We measured aCL in women, aged 15-49 years, to determine if they are an independent risk factor for thromboembolic disease. STUDY DESIGN: Case- control study METHODS: Fifty cases were studied including venous thromboembolism (n=29), stroke and myocardial infarction (n=21), along with 148 age-matched controls. Serum samples were assayed for aCL and anti-beta2 glycoprotein 1 antibodies using the enzyme-linked immunosorbent assay (ELISA). Information on other risk factors was obtained by a standardized questionnaire. RESULTS: aCL were present in 16/50 (32%) of cases compared with 25/148 (17%) of controls (P[?]=[?]0.02). Unadjusted odds ratio (OR) and 95% confidence interval (95% CI) for thromboembolic disease associated with aCL was 2.32 (1.10--4.87). Other risk factors were hypertension, 2.93 (1.20--7.17) and a history of other heart diseases, 12.78 (1.32--123.60). Adjustment for hypertension, diabetes, oral contraceptive use, smoking, alcohol use, varicose veins, a family history of cardiovascular disease and a history of other heart diseases yielded OR (95%CI) 2.99 (1.32--6.80). beta2 glycoprotein 1-dependent aCL were also an independent risk factor, OR 4.56 (1.76--17.83). Subgroup analysis was carried out separately for cases of MI and stroke and for venous thrombosis. Adjusted OR (95% CI) associated with aCL in cases of MI and stroke was 1.76 (0.46--6.73) and 3.32 (1.15--9.54) for venous thromboembolism. CONCLUSION: aCL are a risk factor for thromboembolic disease in young Jamaican women. They confer a strong independent risk for venous thromboembolism. PMID- 11873091 TI - Early short-term intensive cardiac rehabilitation induces positive results as long as one year after the acute coronary event: a prospective one-year controlled study. AB - AIMS: The one-year effects of early and short-term intensive cardiac rehabilitation programmes in patients after acute myocardial infarction or coronary artery bypass surgery (CABG) are not well established. METHODS AND RESULTS: One to four weeks after hospital discharge for acute myocardial infarction (n=55) or CABG (n=54), 109 patients were included in a multidisciplinary ambulatory cardiac rehabilitation programme, lasting 2 to 3 months and including a mean of 33 daily sessions. A complete cardiological assessment of the classical coronary risk factors was performed at entry into the study and again 12 months later, that is 9 to 10 months after the end of the rehabilitation programme. Major effects at one-year follow-up were a high rate of aspirin intake, a low rate of smoking (14% of the patients), a 15% increase in physical capacity, a 7 beats/min decrease in resting heart and a 4 mg/dl increase in the HDL-cholesterol. Body weight increased by 4.9 kg in the patients who stopped smoking; the modest increase in body weight in the other patients reflected a partial weight recovery in the CABG patients. Blood pressure levels also increased at the end of the study but our data in CABG patients and their extrapolation to the post MI patients strongly suggest a progressive return of blood pressure to the pre-acute event levels. In a control group matched for age, sex and type of coronary event, no significant modifications were observed after one year, except for an increase in body weight of 1.7 kg (P < 0.000). CONCLUSIONS: Cardiac rehabilitation which started early after an acute coronary event and regularly followed during 2 to 3 months induced beneficial effects which were still present 9 to 10 months later. Weight gain after smoking cessation was prevalent. The lack of changes in the control group reinforced the benefit of cardiac rehabilitation. PMID- 11873092 TI - Underestimation of the importance of homocysteine as a risk factor for cardiovascular disease in epidemiological studies. AB - BACKGROUND: In epidemiological studies, within-person variability in plasma total homocysteine (tHcy) measurements may dilute the association of 'usual' levels of tHcy with risk of cardiovascular disease, referred to as 'regression dilution'. The aim of this report was to estimate the magnitude of regression dilution after varying intervals of follow-up. METHODS: Regression dilution ratios (RDR) for tHcy were calculated using replicate tHcy measurements obtained after 3, 6 and 8 years from the Rotterdam, Hordaland and Framingham studies, respectively, and after 3, 6, 9 and 12 years from the United Kingdom Prospective Study of type 2 Diabetes Mellitus (UKPDS). RESULTS: The RDR for tHcy decreased with increasing interval in the three population-based studies and in the UKPDS. Moreover, the rate of decline of the RDR in the population-based studies was similar to that obtained in the UKPDS. Using linear regression analysis for the population-based studies, these results suggest an RDR of 0.83 at 2 years, 0.71 at 6 years and 0.53 at 12 years. CONCLUSIONS: These results have important implications for the interpretation of prospective studies of tHcy and cardiovascular disease. Failure to correct for increasing regression dilution using lower RDRs for longer follow up may underestimate the relative risks of cardiovascular disease associated with tHcy by about one-fifth after 2 years and one-half after 10 years. PMID- 11873093 TI - Men of low socio-economic and educational level possess pronounced deficient knowledge about the risk factors related to coronary heart disease. AB - BACKGROUND: The aim of the present study was to determine whether certain background factors such as gender, education and social status were associated with an individual's knowledge of coronary heart disease (CHD) risk factors. DESIGN: A questionnaire survey. METHODS: A questionnaire survey designed to evaluate participants' general knowledge about the risk factors for CHD was used. A total of 1011 50-year-old individuals (457 men and 554 women) from 34 Health Care Centers participated in the study. RESULTS: Knowledge about CHD risk factors was significantly poorer in men than in women. Low education and low socio economic status were other factors related to poor knowledge of CHD risk factors. CONCLUSION: This study showed that men with low educational level and low socio economic status had inadequate information about the risk factors involved in CHD. PMID- 11873094 TI - The multiplicity of atherosclerotic risk factors corresponds to the appearance of increased leukocyte count in the peripheral blood: relevance to the pathogenesis of the disease. AB - White blood cells may have a role in the aetiopathogenesis of atherosclerosis disease in patients with risk factors for this disease. We examined the white blood cell count in a group of 331 patients and controls of the same age group (139 women and 192 men), the numbers of individuals with no, one, two or more atherosclerotic risk factors being 29, 47, 35 and 28 for women and 50, 45, 68 and 29 for men, respectively. The risk factors included were hypertension, hyperlipidaemia, current smoking and diabetes mellitus. A stepwise increment in the white blood cell count was found in both women and men, the respective values for no, one, two or more risk factors being 6.3 +/- 1.5, 7.6 +/- 1.9, 7.5 +/- 1.8, 7.3 +/- 1.4 and 6.6 +/- 1.6, 6.9 +/- 1.9, 7.4 +/- 2.1, 8.1 +/- 2.6 (absolute number of cells per cm x 103). The one-way analysis variance was found to be significant for both women (P=0.01) and men (P=0.01), as well as the entire cohort (P=0.03). We conclude that the multiplicity of risk factors for atherosclerosis is associated with the appearance of an increased number of white blood cells in the peripheral blood. These findings might represent an enhanced inflammatory response in these individuals and at the same time reveal a potential harmful role of the cells in the aetiopathogenesis of the disease. PMID- 11873095 TI - Rosuvastatin demonstrates greater reduction of low-density lipoprotein cholesterol compared with pravastatin and simvastatin in hypercholesterolaemic patients: a randomized, double-blind study. AB - BACKGROUND: Rosuvastatin (Crestor), a new, highly efficacious statin, has demonstrated dose-dependent low-density lipoprotein cholesterol (LDL-C) reductions of up to 65% in a dose-ranging programme with doses of 1 to 80 mg. DESIGN: A randomized, double-blind multicentre trial compared rosuvastatin with commonly used starting doses of pravastatin and simvastatin to determine relative efficacy in LDL-C reduction and impact on other lipid parameters in primary hypercholesterolaemia. METHODS AND RESULTS: A total of 502 patients (greater-than or-equal 18 years; LDL-C greater-than-or-equal 4.14 mmol/l [160 mg/dl] and < 6.50 mmol/l [250 mg/dl] and triglycerides less-than-or-equal 4.52 mmol/l [400 mg/dl]) were randomized to 12 weeks of rosuvastatin 5 mg (n = 120) or 10 mg (n = 115), pravastatin 20 mg (n=]137) or simvastatin 20 mg (n = 130). Rosuvastatin 5 and 10 mg reduced LDL-C by 42 and 49%, respectively, compared with 28% for pravastatin (P < 0.001 versus both rosuvastatin doses) and 37% for simvastatin (P < 0.01 versus rosuvastatin 5 mg; P < 0.001 versus 10[?]mg). National Cholesterol Education Program Adult Treatment Panel II (NCEP ATP II) goals were achieved by 87% of rosuvastatin 10[?]mg patients, 71% of rosuvastatin 5[?]mg patients, 53% of pravastatin patients, and 64% of simvastatin patients; similar proportions of patients achieved NCEP ATP III goals. European Atherosclerosis Society (EAS) goals were achieved by 83, 63, 20 and 50% of patients, respectively. All study treatments were well tolerated. CONCLUSIONS: Both doses of rosuvastatin were more effective than pravastatin and simvastatin in meeting NCEP ATP II and EAS LDL-C targets. Rosuvastatin 10 mg was more effective than pravastatin and simvastatin in meeting NCEP ATP III targets. PMID- 11873097 TI - Esophageal pH monitoring, indications, and methods. AB - Symptomatic gastroesophageal reflux disease (GERD) is a common problem that affects a substantial proportion of the American population. It is estimated that the symptoms of GERD may afflict 40% to 45% of Americans each month. The diagnosis of GERD can be difficult, as its symptoms vary from typical symptoms like heartburn to atypical symptoms such as hoarseness, coughing, and chest pain. Most patients present with typical symptoms and are diagnosed with GERD if they respond to empiric trials of acid suppression. Many tests are available to help with diagnosing GERD in patients who either present with atypical symptoms or who do not respond to acid suppression; however, each test has its own shortcomings. The only test that directly measures whether acid is refluxing into the esophagus is the pH probe, but this test is uncomfortable for the patient, can be difficult to interpret, and may not be necessary in all cases. This article reviews the indications for pH monitoring, its technique, its advantages and limitations, and its role the diagnosis of GERD. PMID- 11873098 TI - Nutritional management of short bowel syndrome in adults. AB - Short bowel syndrome (SBS) comprises the sequelae of nutrient, fluid, and weight loss that occurs subsequent to greatly reduced functional surface area of the small intestine. Signs and symptoms of SBS include electrolyte disturbances; deficiencies of calcium, magnesium, zinc, iron, vitamin B12, or fat-soluble vitamin deficiency; malabsorption of carbohydrates, lactose, and protein; metabolic acidosis, gastric acid hypersecretion; formation of cholesterol biliary calculi and renal oxalate calculi; and dehydration, steatorrhea, diarrhea, and weight loss. Thorough nutritional management is the key factor in achieving an optimal outcome in SBS. Total parenteral nutrition is necessary in the early stages, as is replacement of excess fluid and electrolyte losses. Nutritional management of SBS has traditionally been divided into three phases: an acute phase when total parenteral nutrition is usually begun, an adaptation phase, and a maintenance phase. Recommendations regarding the need for parenteral nutrition vary depending on the presence or absence of certain factors: the ileocecal valve, jejunum, and functional colon. Patients with residual small bowel length of 100 cm or less usually require the administration of parenteral nutrition at home with good results. The total parenteral nutrition diet should consist of a majority of calories from fat, followed by protein, and the remaining as carbohydrates. Vitamins, minerals, and trace elements should also be added accordingly. Although total parenteral nutrition is initially necessary, treatment goals should focus on early transition to enteral nutrition followed by oral feeds. Other recent advances in the medical management of SBS include pharmacologic treatment and the use of specific nutrients and growth factors to stimulate intestinal absorption and adaptation. Both animal studies and clinical trials in humans have shown much promise in supplementation with growth factors and hormones. This strategy is likely to play a greater role in the treatment of SBS in the future. PMID- 11873100 TI - Motility of the transverse colon used for esophageal replacement. AB - The authors studied the motility of transverse colon used for reconstruction of the pharyngogastric transit after esophagectomy. The study included 10 patients who underwent esophagectomy 15 to 201 months (median, 48.5 months) before motility evaluation. Nine patients underwent operation because of caustic injury and one, because of esophageal cancer. The age of the patients ranged from 19 to 54 years (median, 36 years). A manometric esophageal catheter with five side holes spaced 5 cm apart (using the continuous perfusion method) was used to record motility. In three patients, it was not possible to introduce the manometric catheter inside the colon interposition. In the other seven, most of the time there was no contraction when motility was recorded. In four, there was contraction only in the segment 2 to 5 cm below the upper esophageal sphincter. In three, there were peristaltic or simultaneous contractions of long duration, sometimes associated with dry or wet swallows. The motility of colon interposition used to restore transit after esophagectomy is similar to that described for the colon. The contractions may be the consequence of graft distention after successive swallows. PMID- 11873099 TI - Increased prevalence of irritable bowel syndrome in patients with gastroesophageal reflux. AB - GOALS: To determine the prevalence of irritable bowel syndrome (IBS) in subjects with gastroesophageal reflux disease (GERD) compared with non-GERD controls. STUDY: Two hundred subjects were identified from a list of Cedars-Sinai Medical Foundation patients and gastroenterology motility practice subjects with and without a potential diagnosis of GERD. All subjects were then evaluated independently by two blinded physicians who were asked to identify subjects with GERD based on taking a history (gold standard). A follow-up questionnaire was later mailed to patients. This questionnaire included Rome I criteria for IBS. The prevalence of IBS was compared between GERD and non-GERD subjects. Finally, to further strengthen the method, a retrospective review of all subjects' charts was conducted to identify patients who had had 24-hour pH tests, and the prevalence of IBS was determined in this subgroup. RESULTS: Of the 200 subjects, 90 (45%) patients returned the questionnaire. After excluding subjects with IBD and incomplete questionnaires, there were 84 subjects (35 with GERD) included in the analysis. Of the 35 GERD subjects, 25 (71%) were Rome I criteria positive for IBS, whereas only 17 of the 49 (35%) non-GERD subjects had IBS (odds ratio = 54.7, CI = 1.7-13.5, p < 0.01). In 11 of the GERD subjects a 24-hour pH study was available and confirmed GERD. Of these 11 subjects, 7 (64%) met Rome I criteria for IBS. CONCLUSION: There is a higher prevalence of IBS in subjects with GERD compared with subjects without GERD. PMID- 11873101 TI - The inflammatory polyp-fold complex in children. AB - The inflammatory polyp-fold complex (IPFC) is an uncommon endoscopic or radiologic finding in children. In this complex, an inflammatory polyp at the gastroesophageal junction is present, often in continuity with a prominent gastric fold. Histologically, there is an inflammatory infiltrate in otherwise benign gastric and esophageal mucosa. We report four cases of IPFC in children, all associated with reflux esophagitis. In two patients who underwent repeat endoscopy, acid suppression therapy led to a decrease in the size of the polyp and histologic improvement of esophagitis. Four case studies in children with IPFC are presented, followed by a literature review of this endoscopic finding as it applies to children. PMID- 11873102 TI - Magnetic resonance cholangiography versus ultrasound in the evaluation of the gallbladder. AB - Ultrasonography (US) is currently the reference technique for evaluating gallbladder pathology. The aim of this study was to prospectively determine the diagnostic efficacy of magnetic resonance cholangiography (MRCP) in evaluating the gallbladder, as compared with US. The study included 80 patients (mean age, 69.3 years; male-to-female ration, 1.3:1) who underwent prospective US and MRCP; 5 patients in whom MRCP was contraindicated were excluded. In all cases, US was performed before MRCP. Ultrasound was the reference technique for evaluating MRCP sensitivity and specificity. Magnetic resonance cholangiopancreatography provided good image quality in 65 patients (81.2%) and poor image quality in 15 (mostly because of poor patient cooperation). Artefacts did not influence visualization of the gallbladder or evaluation of the background pathology. The sensitivity of MRCP in diagnosing gallbladder stones (43 patients; 97.7%) was comparable to US (44 patients). In contrast, MRCP diagnosed biliary sludge or microlithiasis in 13 patients, versus 5 in the case of US. Magnetic resonance cholangiopancreatography is a good technique for diagnosing cholelithiasis and biliary sludge. However, its high cost, contraindications, and the need for patient cooperation limit the use of the technique in routine clinical gallbladder studies. Magnetic resonance cholangiopancreatography could contribute to the diagnosis of microlithiasis, provided that future studies confirm its greater sensitivity versus US. PMID- 11873103 TI - Increased colonic intraepithelial lymphocytes in patients with Hashimoto's thyroiditis. AB - BACKGROUND: Hashimoto's thyroiditis is an autoimmune thyroid disorder. Lymphocytic colitis and collagenous colitis are characterized by diarrhea with normal endoscopic findings. Autoimmune disorders are common in Hashimoto's thyroiditis and lymphocytic colitis. The aim of this study was to investigate the incidence of lymphocytic colitis in patients with Hashimoto's thyroiditis. STUDY: Fifty patients with well-documented Hashimoto's thyroiditis were included. Twenty patients with nonulcer dyspepsia served as a control group. Five of 50 patients with Hashimoto's thyroiditis reported intermittent diarrhea, whereas no patients had diarrhea in the control group. All patients and the control group underwent total colonoscopy, and multiple colonoscopic biopsies were performed. RESULTS: We found that 40% (20 of 50) of Patients with Hashimoto's thyroiditis had histologic findings consistent with lymphocytic colitis, and one patient in the control group had lymphocytic colitis (p < 0.01). The mean number of intraepithelial lymphocytes was 34.4/100 epithelial cells in these 20 patients, whereas the mean number of intraepithelial lymphocytes was 12.3/100 epithelial cells in the other 30 patients with Hashimoto's thyroiditis (p < 0.05). CONCLUSIONS: There was a higher incidence of histologic findings of lymphocytic colitis in patients with Hashimoto's thyroiditis, although most of the patients were clinically asymptomatic. This finding suggests that lymphocytic colitis may have an asymptomatic clinical course and should encourage further clinical investigations to better anticipate the relationship between autoimmune disorders. PMID- 11873104 TI - Eosinophilic gastroenteritis masquerading as ampullary adenoma. AB - Eosinophilic gastroenteritis is a rare gastrointestinal disorder of undetermined etiology that is characterized by eosinophilic infiltration of the gut wall. The presenting symptoms depend on the site and depth of intestinal involvement and varies from nausea, vomiting, and abdominal pain to acute bowel obstruction. Pancreaticobiliary obstruction caused by eosinophilic gastroenteritis is rare. We report a 39-year-old man who presented with abdominal pain, vomiting, abnormal liver tests, and a duodenal mass on upper endoscopy. Blood tests showed peripheral eosinophilia. Abdominal computed tomography scan showed a suspected mass in ampullary region. At endoscopic retrograde cholangiopancreatography, both pancreatic and common bile duct were dilated with no obvious ductal strictures. Biopsies from the duodenal mass showed evidence of eosinophilic gastroenteritis. He was successfully treated with prednisone, and his liver test results returned to normal. In conclusion, this unusual case of eosinophilic gastroenteritis presented with duodenal mass that was masquerading as an ampullary adenoma causing pancreaticobiliary obstruction. PMID- 11873105 TI - Cytomegalovirus enterocolitis in an immunocompetent individual. AB - We report a rare case of cytomegalovirus (CMV) enterocolitis in a healthy 57-year old woman. In March 1999, she developed hematochezia, diarrhea, and abdominal pain. Total colonoscopy on March 17th showed multiple aphthoid lesions and friable mucosa from the terminal ileum to the rectum and a shallow ulcer on the ileocecal valve. Repeat total colonoscopy on April 19th showed faded aphthoid lesions in the terminal ileum, and biopsy specimen revealed CMV inclusion bodies. Symptoms and endoscopic findings improved without any specific medication. In previous reports, the definition of "immunocompetent individual" varied. Here, we define immunocompetent individual as one who has no associated diseases, is not under immunosuppressive therapy, has no recent history of operation, is negative for human immunodeficiency virus antibody, is not pregnant, has no obvious infectious course, and is less than 70 years of age. This is the ninth report of CMV enterocolitis in an immunocompetent individual in the world literature. PMID- 11873106 TI - Constrictive pericarditis in chronic ulcerative colitis. AB - Acute pericarditis has been described as an extraintestinal manifestation of inflammatory bowel disease (IBD), as well as a consequence of IBD treatment, specifically sulfasalazine and mesalamine. Until now, there have been no reported cases of constrictive pericarditis associated with IBD or its treatment. A 37 year-old woman with a 24-year history of chronic ulcerative colitis (CUC) presented with a 3-month history of fevers, palpitations, dyspnea, syncope, and retrosternal chest pain. Two weeks before symptoms, she had initiated oral mesalamine for an ongoing CUC flare. Physical examination suggested constrictive pericarditis. An echocardiogram revealed a thickened pericardium with a nearly circumferential fibrinous effusion, with Doppler confirming diastolic compromise. The patient proceeded to radical pericardectomy. Pathological examination showed grossly hemorrhagic acute and chronic pericarditis, with cultures and cytology negative. To date, only 104 cases of IBD with acute pericarditis have been reported, with fewer than 10 cases of mesalamine-induced acute pericarditis reported. This is the first reported case of constrictive pericarditis related to IBD or its treatment. Although our patient may have had IBD-associated constrictive pericarditis, her mesalamine use raises the possibility of a drug induced constrictive pericarditis. PMID- 11873107 TI - An unusual presentation of enterocolic lymphocytic phlebitis. AB - Enterocolic lymphocytic phlebitis (ELP) is a recently described disease characterized by phlebitis in the wall and mesentery of the colon and small bowel. Unlike other systemic or localized vasculitic diseases that can have similar gastrointestinal manifestations, there is no involvement of the arterial system or evidence of systemic vasculitis. The lymphocytic phlebitis affects not only the grossly involved intestinal segment but also the apparently healthy bowel. The diagnosis of ELP is histologic, and other systemic vasculitis must be ruled out. Surgical resection of the bowel not only provides the diagnostic tissue but also leads to resolution of the symptoms. In the literature, ELP has most commonly been reported to present as an acute abdomen. We describe a case of ELP presenting as a large abdominal mass, without evidence of an acute abdomen. This mass was actually diffusely thickened colonic serosa, caused by extensive organizing fat necrosis and marked edema of the serosa and bowel wall. PMID- 11873108 TI - Update on nonalcoholic fatty liver disease. AB - Nonalcoholic fatty liver disease is now recognized as the most common liver disease in the United States, with a prevalence of approximately 5% in the general population and up to 25% to 75% in patients with obesity and type II diabetes mellitus. Nonalcoholic fatty liver disease is a clinicopathologic syndrome with a wide spectrum of histologic abnormalities and clinical outcomes. Hepatic steatosis has a benign clinical course. In contrast, nonalcoholic steatohepatitis (NASH) may progress to cirrhosis and liver-related death in 25% and 10% of patients, respectively. Cases occur most commonly in obese, middle aged women with diabetes. However, NASH may also occur in children and normal weight men with normal glucose and lipid metabolism. The pathophysiology involves two steps. The first is insulin resistance, which causes steatosis. The second is oxidative stress, which produces lipid peroxidation and activates inflammatory cytokines resulting in NASH. Liver biopsy provides prognostic information and identifies NASH patients who may benefit from therapy. Treatment consists of managing the comorbidities: obesity, diabetes, and hyperlipidemia. Although antioxidant therapy with vitamin E is often used, ursodeoxycholic acid is the only drug that has shown benefit and is the most promising of the drugs currently being investigated. Future therapies will depend on a greater understanding of the pathophysiology and should focus on diminishing fibrosis. PMID- 11873109 TI - Hepatic Fas protein expression might be a predictive factor for hepatocellular carcinoma development in patients with chronic hepatitis C undergoing interferon therapy. AB - BACKGROUND: Several studies have revealed that interferon treatment may reduce the incidence of hepatocellular carcinoma (HCC) in patients with chronic hepatitis C virus (HCV). However, even after eradication of HCV, patients remain at risk for developing HCC. STUDY: Of 153 consecutive HCV patients who were treated with interferon and followed up for 5 years, 17 (11.1%) developed HCC. To elucidate predictive factors of HCC development, multivariate analysis was done for the 153 patients, and Fas protein expression in the biopsied specimens of liver before interferon treatment was examined in 17 patients who developed HCC and 17 patients who did not. RESULTS: Among the independent factors (sex, age, HCV genotype, HCV-RNA level, effect of interferon therapy, serum alanine aminotransferase before interferon therapy, and histologic stage and grade) tested by Cox proportional-hazards analysis, histologic stage (hepatic fibrosis) before interferon was significantly associated with HCC development (p = 0.01). In addition, the intensity of Fas protein expression was significantly greater in the liver specimens of the patients who developed HCC than in those who did not (p = 0.015). CONCLUSION: Histologic stage (hepatic fibrosis) and Fas protein expression before interferon treatment might be indicative of the need for intensive follow-up in patients with chronic hepatitis C undergoing interferon therapy. PMID- 11873110 TI - A descriptive evaluation of eligibility for therapy among veterans with chronic hepatitis C virus infection. AB - GOAL: To assess the number of chronic hepatitis C patients eligible for therapy. BACKGROUND: Recent studies have shown improved response rates to treatment of chronic hepatitis C infection. However, treatment with interferon alfa has major side effects, and many patients may not be eligible for therapy. STUDY: One hundred consecutive patients with positive hepatitis C serologies at the Durham Veterans Affairs Medical Center were evaluated. Medical records were reviewed, and the patients were interviewed. Patients were considered ineligible for therapy if they had severe mental illness, hazardous alcohol consumption, current drug abuse, decompensated cirrhosis, dementia, terminal illness, diabetic ketoacidosis, and severe cardiac or pulmonary disease or if they were homeless. RESULTS: Of the 100 patients, 92% were male and 51% were African American. The mean age was 47.3 +/- 5.6 years. Only 32 of the 100 patients were eligible for therapy. Hazardous alcohol consumption was present in 44%. Major depressive symptoms were present in 12%. CONCLUSIONS: The minority of chronic hepatitis C patients were eligible for therapy. Significant rates of hazardous alcohol consumption and psychiatric disorders were present. For these patients to complete or become eligible for therapy, a multidisciplinary approach with psychiatric and substance abuse treatment will be necessary. PMID- 11873111 TI - Huge adenomatous hyperplasia of the liver. AB - Adenomatous hyperplasia (AH) of the liver is defined as a regenerative overgrowth with limited growth potential. Patients with AH of the liver usually have cirrhosis of the liver as well. Adenomatous hyperplasia is also described as a benign nodule more than 8 mm, which is the main differentiation between AH and regeneration nodules (which are less than 8 mm). Adenomatous hyperplasias more than 20 mm is extremely rare in the clinicopathologic studies. We present two cases of extraordinarily large AH (one was 100 mm and the other, 30 mm). Both patients were alcoholic, and one also had viral hepatitis B. By clinical, biochemical, and upper gastrointestinal endoscopic examinations, we diagnosed liver cirrhosis in both. Sizable nodules were discovered in their livers using imaging studies (including ultrasonography, computed tomography, magnetic resonance imaging, and positron emission tomography), and percutaneous liver biopsies of the nodules showed their cirrhotic background. However, neither of the patients developed hepatocellular carcinoma during the follow-up period. PMID- 11873113 TI - Absence of the common bile duct and junction of the cystic duct with the left hepatic duct in a patient with chronic portal vein thrombosis. AB - A50-year-old male patient was admitted to the hospital because of fatigue and a palpable abdominal mass. The diagnoses of chronic renal failure and portal vein thrombosis were established by specific investigations. Incidentally, junction of the cystic duct with the left hepatic duct and absence of the common bile duct were found during endoscopic retrograde cholangiopancreatography study. To the authors' knowledge, this is the first reported case of an absence of the common bile duct and junction of the cystic duct with the left hepatic duct in a patient with chronic portal vein thrombosis. Such lesions may cause unexpected complications during biliary surgery. PMID- 11873112 TI - Hepatic hydrothorax associated with vitamin a toxicity. AB - We report the first case of an adult presenting with respiratory symptoms caused by hepatic hydrothorax secondary to vitamin A intoxication. The patient was a 52 year-old woman who presented to the hospital with progressive dyspnea. Evaluation demonstrated mild elevation of her liver function tests, ascites, and a right pleural effusion. The patient consumed a variety of vitamins, including vitamin A. Her estimated vitamin A intake was at least 162,300,000 international units (IU) during 18 years. She dramatically escalated her dose the year before admission for a total acute dose of 98,550,000 IU, with a daily intake of 270,000 IU. The recommended daily allowance is 4,000 IU. A transjugular liver biopsy revealed histopathologic changes consistent with vitamin A toxicity: hypertrophy and hyperplasia of hepatic stellate cells, focal pericellular fibrosis, mild perivenular fibrosis, and minimal, predominantly microvesicular steatosis. Despite the absence of cirrhosis, pressure readings demonstrated portal hypertension. During her hospitalization, the patient's symptoms and biochemical profile improved. As the large and generally unregulated United States dietary supplement industry continues to grow, it is increasingly likely that individuals will present with the signs and symptoms of vitamin excess rather than vitamin deficiency. Physicians need to remain alert to the varied presentations and toxic manifestations of excessive vitamin use. PMID- 11873114 TI - Rapid placement of the Sengstaken-Blakemore tube using a guidewire. PMID- 11873115 TI - Gastric epithelial dysplasia and gastric cancer in young dyspeptic patients treated with proton pump inhibitors. PMID- 11873116 TI - Acute pancreatitis caused by ketorolac tromethamine. PMID- 11873117 TI - Gastric antral vascular ectasia causing severe hypoalbuminemia and anemia cured by antrectomy. PMID- 11873118 TI - RCAS1 expression in immune-mediated liver diseases. PMID- 11873120 TI - Regarding "Celiac disease is highly prevalent in lymphocytic colitis". PMID- 11873119 TI - Leptin response in patients undergoing interferon therapy. PMID- 11873122 TI - Small bowel obstruction as a unique complication of squamous cell carcinoma of the penis. PMID- 11873126 TI - Multicultural Realities: A Special Number of 43: Number 4, Fall 1999. PMID- 11873125 TI - Review of Psychiatric Series, Vol. 19. PMID- 11873199 TI - A challenge to the new government: be radical. PMID- 11873200 TI - Health visitors are nurses, but health visiting is not nursing. PMID- 11873201 TI - The benefits of integrated nursing teams in primary care. AB - This article reports a study in which a literature search, questionnaire study and interviews were used to measure the costs and benefits to patients and professionals when nurses work in integrated nursing teams in primary care. Our study showed that there are significant benefits for the nurses' employers, the primary health-care team, the nurses and the patients. Compared with traditional models, delivery by integrated nursing teams can offer a much higher quality service. Patients can expect to be cared for by a team in which morale is high, communication is good and care is efficient and effective. The team can also offer many specialist services that were not previously available. Integrated nursing teams are most effective when the management savings are used by the teams to meet their professional and client needs. Financial savings, therefore, cannot be expected. The difficulties that arise when such teams are set up should be short-term and surmountable, but will be influenced by the motivation of the team members and the efficiency of the human resource management. PMID- 11873203 TI - Restoring pleasure: nutritional management of dysphagia. AB - Dysphagia is associated with many different conditions and although widespread it is frequently not recognized. People with dysphagia are at risk of developing serious complications such at malnutrition, dehydration and aspiration. Texture modification, food fortification and dietary supplements are important elements in effective nutritional management of dysphagia and can also help bring back some of the pleasure to mealtimes. The ideal approach to the management of dysphagia involves a multidisciplinary team. PMID- 11873202 TI - Cost effectiveness in leg ulcer management in the community. AB - Wound care and leg ulcer management account for the largest part of district nurses' workload. This has major implications on prescribing budgets of primary care groups (PCGs). Ineffective care results in an expensive waste of financial resources and poor quality care for the patients involved. Two case studies of patients with venous leg ulcers are used as an example of how practice can vary within one PCG. Continuing support and training is needed for all community nurses to enable them to meet national and local standards and guidelines. PMID- 11873204 TI - Geriatric assessment in primary care: formulating best practice. AB - Comprehensive geriatric assessment (CGA) is a structured approach to measuring physical, mental and social functioning of older people to identify needs and to plan care. Meta-analysis of trials of CGA suggest that it is cost-effective, but there is no agreed approach to its implementation in primary care. Our aim was to develop a best-practice model for geriatric assessment in primary care. We took an iterative approach to development, combining expert and local stakeholder opinion, and using semi-structured interviews to assess patient and practitioner experience in nine general practices in Sheffield. Patients were aged 75 and over, living at home. The best-practice model was the use of a standardized instrument (EASY-Care) to unselected patients aged 75 years and over living at home or in residential care, administered by a practice nurse in the context of an over-75s health check. There was high patient and practitioner acceptability, and significant cost savings were noted. Key beneficial features were the assessment of mental health and sources of support; goal-setting; generation of a disability score; and high patient satisfaction from contact with nursing staff. We conclude that geriatric assessment in primary care is feasible, economical and beneficial to patients and practitioners. Nursing staff are central to successful implementation of geriatric assessment in primary care. PMID- 11873205 TI - Medicines and older people: a nurses' guide to administration. AB - Older people represent only 18% of the population but receive approximately 45% of all prescribed medication in the UK (Offerhaus, 1997). Problems arising from drug therapy are numerous, as medication will often act in a number of complex and even surprising ways. As the number of older people grows, it becomes increasingly necessary for nurses to recognize and understand the legal and professional issues relating to the administration of medicines. This article outlines some of the potential problems confronting the nursing profession and offers some practical solutions to common dilemmas. PMID- 11873206 TI - Assess, negotiate, treat: community prescribing for chronic wounds. AB - The cost of prescribing wound-care products in general practice in 1999 exceeded 95 million (Department of Health (DoH), 1999a). However, there is a lack of randomized controlled trials to assess the effectiveness of wound-care products (National Prescribing Centre, 1999a) and there is evidence that non-effective treatments are being prescribed (DoH, 1999a). Use of the prescribing pyramid, as taught during nurse prescribing courses, may help community nurses justify their decisions and assess their wound-care practices. Wounds commonly encountered in the community include pressure sores, leg ulcers, fungating wounds and cavity wounds. The ideal choice of product is one that allows moist wound healing, is cost effective, clinically effective and acceptable to the patient. PMID- 11873207 TI - Transfix: a new range of all-silicone male incontinence sheaths. AB - Urinary incontinence can have a significant detrimental effect on a person's body image and self-esteem because it undermine society's norms relating to control of the body. This can be further complicated by the use of devices to control urinary incontinence. Sheath drainage systems are the most common method of managing urinary incontinence in men. A discreet, reliable sheath drainage system can have both physical and psychological benefits. A full continence assessment is paramount for a successful outcome as there are several potential problems associated with this method of management. There are many products available, so the health professional needs a comprehensive knowledge of both products and application techniques. The Sims Portex Transfix range of all-silicone self adhesive sheathes have many benefits and offer a reliable form of sheath management for male urinary incontinence. PMID- 11873208 TI - Intermediate care doesn't provide all the answers. PMID- 11873209 TI - Use of the Polaroid Macro 3 SLR camera in wound assessment. PMID- 11873210 TI - Nurse shortage crisis will continue unless action is taken. PMID- 11873211 TI - The law relating to teenage pregnancy needs to be clearer. PMID- 11873212 TI - Psychiatric nurse who was too busy to keep up-to-date records. PMID- 11873214 TI - Management of intravascular devices to prevent infection. AB - Intravenous therapy is an essential part of clinical care used in a wide variety of healthcare settings. Thus, intravenous catheters have become indispensable to clinical practice. However, catheter-related-bloodstream infections (CR-BSIs) are a major source of morbidity and mortality, especially in hospital patients. Strategies to prevent infection need to change and develop to reflect the advances in technology and delivery of health care. Current guidance recommends a number of strategies to reduce the incidence of CR-BSIs. These include applying the principles of asepsis, the choice of catheter material, the site of insertion and when to replace equipment used. This article reviews the guidelines for current clinical practice. PMID- 11873213 TI - Collecting pressure ulcer prevention and management outcomes: 1. AB - This article, the first of two parts, presents the argument that a combination of efficacy and effectiveness is required to assess fully the impact of interventions such as pressure-redistributing beds and mattresses. The methodology adopted within a multinational, multicentre, prospective, non randomized cohort study, designed to record the occurrence and characteristics of patients vulnerable to, or with, established pressure ulcers, is described. General demographic data and the characteristics of the pressure ulcers experienced by the 2507 UK subjects recruited to the study across four UK hospitals between July 1996 and May 1998 are presented, with pressure ulcers affecting 218 subjects of whom 100 presented with ulcers on admission to hospital. Fourteen subjects developed severe ulcers, while a further 24 were admitted with full-thickness pressure ulcers. The second part of this article will report further details of the characteristics of the UK sample. Future articles will consider the subjects examined in greater detail and will also discuss the combination of this dataset with similar data collected in the USA. PMID- 11873215 TI - Contemporary issues in the care of sick children and their families. AB - This article considers the response of children's nurses to contemporary change in the management of sick children and their families throughout the last decade. In recognizing the rich and diverse history of children's nursing the impact of those factors which have led to the development of operational strategies to manage children's services will be considered. The quest, not always successful, to coordinate care as a seamless web of services to children and their families will be investigated, as will the role dimensions encompassed by specialist children's nurses and their interface with interprofessional colleagues. A range of nurse-led services in a variety of contemporary child health settings will be explored. PMID- 11873216 TI - Pregnant minors: confidentiality issues and nurses' duties. AB - Abortions in those under 16 years of age raise the issue of what to do in the face of a request that the young person's parents not be involved. The first question in such cases is whether or not the young person is competent to request confidentiality. A younger person who is competent is owned the same duty of confidentiality as an adult. In practice this means that some such requests can be granted straightforwardly. However, in many cases the teenager's pregnancy raises concern about child abuse. In the face of a serious crime the obligation to respect confidentiality is overridden. This creates a dilemma. We cannot respect confidentiality fully in such cases, but a policy of failing to do so may lead young people to seek (illegitimate) help elsewhere. Therefore, reform of the current system may be needed. PMID- 11873217 TI - Physiological insult/injury: pathophysiology and consequences. AB - This article provides a review of the pathophysiological responses of the body to an insult. Once the initial insult has occurred, common pathways are identifiable: the inflammatory immune response (IIR), the neuroendocrine system is stimulated, and endothelial damage initiates the clotting cascade, all of which play an important role in maintaining haemodynamic normality and promote healing. However, after an injury/insult patients are exposed to treatments, invasive procedures and may develop complications which can stimulate further these physiological processes. This can lead to alterations in oxygen supply and demand, metabolism and to the distribution of circulating volume. In some situations this can lead to overstimulation of the processes which, instead of promoting healing, cause damage. Why this downward progression and overstimulation occurs in some patients, despite optimum efforts at treatment, are not fully understood, but can lead to serious organ damage and even death days or weeks after the insult. PMID- 11873218 TI - Job satisfaction of registered nurses working in an acute hospital. AB - The links between job satisfaction and the retention of nurses are well established in the literature. Given the current difficulties experienced by the NHS in recruiting and retaining sufficient numbers of nurses, the need to understand how satisfied nurses are in their jobs becomes clear. This study set out to measure the job satisfaction of registered nurses working in adult acute medical and surgical specialties using the Mueller McCloskey (1990) Satisfaction Scale, a 31-item Likert questionnaire. Data were collected from 141 E-grade nurses working in a ward environment. In addition to providing a general outline of reported job satisfaction, the findings of this study identify key areas of reported job satisfaction around peer socialization and dissatisfaction with childcare facilities and control over work conditions. Based on these findings, recommendations are made that may affect positively nurse retention and suggestions put forward for future use of the Mueller McCloskey Satisfaction Scale. PMID- 11873219 TI - Legal aspects of consent 22: nurses' position when obtaining consent. AB - Case scenario: staff nurse Fawn was asked to take on the role of getting the patients to sign the consent forms before surgery was carried out. She was concerned at the legality of her being asked to undertake this since she would not be carrying out the operations. What is the law? PMID- 11873221 TI - Uncertainty in the next era of professional regulation. PMID- 11873220 TI - Clinical efficacy of Comfeel Plus Transparent Dressing. AB - Hydrocolloid dressings have been in use for more than 20 years. They are indicated for the treatment of granulating, superficial wounds with low to moderate exudates. There are a number of hydrocolloid dressings available, one of which is Comfeel Plus Ulcer Dressing. This differs from the others by combining a hydrocolloid with calcium alginate in its formation to increase the absorptive capacity. This article examines another product of the Comfeel Plus range, Comfeel Plus Transparent Dressing (Coloplast A/S) and describes a small evaluation of the product. The conclusion is that Comfeel Plus Transparent Dressing performs well in terms of healing, compliance, adherence and flexibility. Furthermore, it is transparent and it is therefore possible to follow the wound healing process without removing the dressing. PMID- 11873222 TI - A&E nurses must not let themselves be used. PMID- 11873223 TI - NICE guidelines unfair for Alzheimer's disease. PMID- 11873224 TI - Case 59: absenteeism from work. A&E nurse who was absent from her unit and patients. PMID- 11873225 TI - Recognizing the feet as being at risk from pressure damage. AB - This article reports the findings of a survey and an audit undertaken to investigate the provision of foot support in a university teaching hospital. Phase I surveyed strategies employed to support feet and phase II audited the use of the Repose Foot Protector, manufactured by Frontier Therapeutics, specifically designed to provide pressure support for the feet. Patients with reduced mobility, nursed out of bed in a chair, have been highlighted as a group potentially at risk of tissue damage to the heels. This survey of current strategies employed to support feet included 289 patients. Patients included were from both hospital and community settings. The survey reported a lack of specialist equipment for the heels of patients with reduced mobility sitting in a chair. Only 67 (23.2%) patients were allocated foot support (typically a stool, with or without a pillow) to use while seated out of bed in a chair. The audit of requests for a new device to protect feet included 100 patients. The main reasons for requesting this device included pressure relief (81 occasions), to treat 'foot drop' (32 occasions) and in promoting comfort (31 occasions). There was a significant improvement in the skin condition of the heels and comfort (P<0.0001) from study entry to exit. This audit indicated a high level of both staff and patient satisfaction. PMID- 11873226 TI - Legal aspects of consent 16: statutory provisions and living wills. AB - Case Scenario: Mary had cared for her invalid mentally infirm mother for many years and was determined that if she ever lost her own mental capacity she would not wish to be kept alive. She drew up a document, witnessed by her sister, that stated in the event of her suffering from any form of mental incapacity she would not wish to be fed or have any medical or nursing intervention. Some years later she began to suffer from the early signs of Alzheimer's disease. Her sister had died, but staff were aware that Mary still carried this living will on her person. She is now refusing all food. What is the law? PMID- 11873227 TI - Death of adolescents: parental grief and coping strategies. AB - The death of an adolescent is a particularly complex issue. The process of grieving and coping can be complicated by the tension that may have existed in the parent/child relationship because of the conflict in terms of personal ideology at this stage in the adolescent's development. As a result, parents of adolescent children who die have the potential to experience abnormal grief reactions. Parental coping strategies vary according to the mode of death and outlook adopted by the parents, although some studies suggest parents bereaved of adolescent children do not exhibit marked difference in grieving or coping in comparison to other groups of parents. Healthcare professionals must recognize the specific and special needs of this parent group, and provide appropriate support to minimize the risk of harmful sequelae that may occur as a result of inappropriate and insensitive care. PMID- 11873228 TI - Child health nurses' perceptions of enquiry-based learning. AB - Since the publication of Fitness for Practice (UKCC, 1999), most preregistration nursing curriculae embrace the principles of enquiry-based learning (EBL) as a method of educating students. EBL uses genuine real-life client scenarios which provide the students with an opportunity to explore a range of issues directly pertaining to client care in a variety of contemporary nursing settings (Long et al, 1999). This evaluative study of one group (n=12) of child branch students conducted during the last EBL session of the course reflects the students' experiences of this method of teaching over a 3-year period. Nominal group technique (Delbecq and Van de Ven, 1971) was utilized to give a quantitative dimension to the reporting and recording of individual student reflections of EBL. The results demonstrate overall satisfaction with this method of student learning. A number of concerns raised by students reflect some of the pitfalls associated with EBL and these have important ramifications for nurse educators and clinical placement mentors. However, the small size limitations of this study do not allow for generalizability. PMID- 11873229 TI - National Service Framework for Older People: management of falls. AB - The National Service Framework (NSF) for Older People was launched in England in the Spring of 2001. It sets out national standards and guidelines and with an increasingly ageing population is welcomed as a way forward to improving the care of older people across the health and social care spectrum. In Wales, work is underway in developing an 'Older persons' strategy', which is expected to reflect the main principles of the NSF. The NSF is underpinned by eight standards which are identified as priority issues. This article focuses on standard six: the prevention, treatment and management of falls. The implications and opportunities for health promotion that may arise from the standard are considered, in particular with reference to the development of an integrated falls service. PMID- 11873230 TI - Patient participation in peer support groups after a cardiac event. AB - Peer support groups may be important in long-term rehabilitation after a cardiac event. Questions can be raised about the clients that peer support groups attract. The aim of this study was to compare people who chose to attend peer support groups after a cardiac event with people who declined to attend with regard to health conditions, personal traits, lifestyle and available social support. Patients who sustained a myocardial infarction or were treated with percutaneous transluminal coronary angioplasty or coronary artery bypass surgery (n = 197) responded to a questionnaire. The results showed that there were differences between attenders (n = 64) and non-attenders (n = 133). Attenders reported more health problems than non-attenders, but scored higher on several dimensions of social support. Non-attenders, however, seemed to have a somewhat more relaxed attitude to life than attenders. The homogeneity of the subjects made it difficult to discern which personal and situational factors were of importance for social support seeking in peer support groups. The research indicates that innovative approaches are needed to encourage participation in existing peer support groups. PMID- 11873231 TI - Tempur-Med: choosing the correct pressure area care system. AB - The prevention and treatment of pressure ulcers is a major concern for hospital nursing staff. Selection of the appropriate equipment for pressure area care can prove difficult, as a great many different systems are available. Tempur-Med provides a simple and cost-effective solution to this problem with a range of products ensuring continuity of care in all areas of the hospital. PMID- 11873232 TI - Is preregistration nurse education being ignored? PMID- 11873233 TI - Collaboration: a critical challenge. PMID- 11873234 TI - Malignant cutaneous wounds: developing education for hospice, oncology and wound care nurses. AB - This article reports the results of an educational project, which included a needs assessment survey and the development of a malignant cutaneous wound educational programme. As there is little information on the differences between the management of malignant cutaneous wounds in the hospice and oncology settings, a survey was undertaken to identify differences encountered by hospice and oncology nurses. This needs assessment survey also included enterostomal/wound-ostomy-continence (ET/WOC) nurses who serve as wound care consultants in oncology and hospice settings. The 12-question survey addressed the frequency of wounds, symptoms, and psychosocial issues encountered. The education programmes was individualized to the setting by determining the differences between the nurses' roles based partially on the survey information. One interesting survey finding was that many of the nurses were using the internet to obtain answers for wound management questions; therefore, a future implication of this project is the need to develop an online malignant cutaneous wound educational programme for nurses. PMID- 11873235 TI - Spirituality in palliative care: what language do we need? AB - Spirituality can be described as a search for meaning. At the time of a significant life crisis, such as is experienced by patients in palliative care, this search can become more pertinent. Patients and their families describe times of questioning and loss of faith as well as discovery, growth and a deepening of experience to a place of inner peace. Because spirituality has an increasingly wide range of interpretations, the delivery of spiritual care can understandably be equally diverse. This article has its limitations in that it concentrates on Christianity, partly because it is a known starting place from the experience of the author and still is relevant for many patients in the West. The holistic/whole person approach to nursing care has helped to refocus on the spiritual dimension of care. However, nurses require support and guidance as to how to approach spiritual care and understand its concept, which appears resistant to language -- their basis for communication. PMID- 11873236 TI - Should we give palliative care to all those that need it? PMID- 11873237 TI - Improving the management of breathlessness using a clinical effectiveness programme. AB - Breathlessness represents a significant problem for the person with advanced cancer. Uncontrolled breathlessness ranks highly in terms of uncomfortable symptoms experience, causing pain and distress to the patient and resulting in significant anxiety to their carers. The key to the provision of effective care lies in the informed application of the nursing process, underpinned by a sound knowledge base in relation to the nursing management of breathlessness. Theoretical knowledge enables nursing staff to offer appropriate interventions for the management of breathlessness in collaboration with other members of the multidisciplinary team. This article will discuss the cause and management of breathlessness in the person with advanced cancer; the discussion focuses on the application of research-based interventions and the evaluation of clinical outcomes in a UK clinical governance context. PMID- 11873238 TI - Nurses' attitudes to palliative care in nursing homes in Western Australia. AB - Nursing homes are one of the care settings in Western Australia where older people may spend their final years. Residents should be able to receive palliative care where appropriate, but this type of care is not always available at some nursing homes in the state. This study investigated nurses' attitudes to palliative care in nursing homes by examining their cognitive, affective and behavioural information. A sample of 228 nurses working in nursing homes completed a questionnaire, using a free response methodology. Results showed that participants had either a positive or negative attitude to palliative care. Cognitive and affective information significantly and independently predicted the attitudes of nurse, whereas knowledge of palliative care did not contribute significantly to these attitudes. Nurses currently working in palliative care were more positively disposed towards such care, but this disappeared when they ceased working in the area. There is an emphasis on education in the literature which does not take into account the beliefs and emotions of the nurse. Therefore, there is a need to consider these in undergraduate and postgraduate training for nurses. Current experience is also important in palliative care education. The results obtained from nurses in this study should be incorporated into policy for introducing palliative care into nursing homes and used to provide support and assistance to nurses working in this field. PMID- 11873239 TI - Effects of interleukin 2 therapy on lymphocyte magnesium levels. AB - Interleukin 2 (IL-2) can cause partial or complete tumor regression in approximately 20% of patients with renal cell carcinoma. Among the many physiologic effects of IL-2, decreased serum levels of the divalent cations magnesium (Mg) and calcium have been demonstrated, with corresponding decreases in their urinary excretion. We investigated the effect of IL-2 on lymphocyte Mg levels among patients receiving three different dosing regimens. Twenty-eight patients with metastatic renal cell carcinoma were treated with high-dose intravenous, low-dose intravenous, or subcutaneous IL-2 therapy. Serum ionized Mg, urinary Mg, and peripheral blood mononuclear cell Mg levels were measured in samples from patients during treatment and compared with pretreatment levels. Serum Mg and ionized Mg levels decreased for all patients within 12 hours of treatment (P <.005) and remained low for the duration of therapy. Urinary Mg decreased in parallel with serum levels in all patients (P <.005). The peripheral blood mononuclear cell Mg content per cell increased within 24 hours of treatment (P <.005). The magnitude of these changes was similar during the first week of treatment for patients receiving intravenous or subcutaneous administration of IL 2. During IL-2 therapy, lymphocyte Mg increases coincident with serum Mg depletion. Mg availability may have functional implications for lymphocyte proliferation and integrin function. PMID- 11873240 TI - Regulation of apoptotic cell death by cytokines in a human salivary gland cell line: distinct and synergistic mechanisms in apoptosis induced by tumor necrosis factor alpha and interferon gamma. AB - We examined the mechanisms of apoptosis in a human salivary gland (HSG) cell line induced by tumor necrosis factor (TNF) alpha and interferon (IFN) gamma. DNA fragmentation and the activation of caspase-3 were determined in HSG cells cultured with TNF-alpha or IFN-gamma. Mitochondrial dysfunction also appeared to be involved in the process because a disruption of mitochondrial transmembrane potential with the activation of caspase-9 was demonstrated in TNF-alpha- and IFN gamma-stimulated HSG cells. Activation of caspase-8 was thought to be essential in TNF-alpha--induced apoptosis of HSG cells; however, the activation of caspase 8 was not involved in IFN-gamma-induced apoptosis of HSG cells. In contrast, Bcl 2 appeared to be an indispensable regulatory molecule in IFN-gamma-induced, but not in TNF-alpha-induced, apoptosis of HSG cells because its expression was inhibited in IFN-gamma-stimulated, but not in TNF-alpha-stimulated, cells. The inhibitory effect of IFN-gamma in Bcl-2 expression was enhanced by coadministration of TNF-alpha and, interestingly, apoptosis of HSG cells, as assessed by DNA fragmentation and the activation of caspase-9 and caspase-3, and disruption of mitochondrial transmembrane potential was also synergistically augmented by TNF-alpha and IFN-gamma. Our results suggest that cytokines expressed in the salivary glands of patients with Sjogren syndrome play an important role in regulating apoptosis of acinar-ductal epithelial cells through distinct and synergistic mechanisms, thereby modulating salivary gland function in patients with Sjogren syndrome. PMID- 11873241 TI - Lysophosphatidic acid augments fibroblast-mediated contraction of released collagen gels. AB - Lysophosphatidic acid (LPA) is a glycerophospholipid released from platelets that has multiple biologic effects. The present study evaluated the potential of LPA to modulate tissue repair and remodeling by modifying human lung fibro-blast mediated contraction of three-dimensional collagen gels. The contraction of native collagen gels caused by human fetal lung fibroblasts was augmented by LPA in a concentration-dependent manner. The estimated median effective concentration was 3 x 10(-7) mol/L, which was well below the concentrations likely released by platelets in tissues. LPA-augmented contraction was not blocked by pertussis toxin or cholera toxin but was inhibited by inhibition of phospholipase C. Neither calcium mobilization nor protein kinase C appeared to play a role. In contrast, the effect of LPA appeared to depend on a kinase inhibited by staurosporine but not by genistein or GF109203X, suggesting a process that depends on phospholipase C and may involve a novel protein kinase. By modulating fibroblast-mediated remodeling, LPA could play a role in the tissue remodeling that characterizes wound repair. PMID- 11873242 TI - Role of bile mucin in bacterial adherence to biliary stents. AB - Biliary stent placement is a well-established method of relieving obstructive jaundice. However, a frequent complication is occlusion of the stent caused by bacterial biofilm formation and sludge accumulation. In this study we investigated the possible effect of bile mucin on bacterial adherence to biliary stents at the initial stage of biofilm formation. By means of an in vitro bile perfusion system, polyethylene stents were perfused with pig gallbladder bile infected with Escherichia coli. The concentrations of mucin in the pig bile were adjusted with purified mucin. The amount of bacteria adhering to the inner surface of the stents was measured and compared for stents perfused with bile containing various concentrations of mucin. Furthermore, we conditioned the stent inner surface with purified pig bile mucin and observed the effect of the conditioning on subsequent bacterial adherence. In addition, a common method for assaying bacterial adhesion with polystyrene microtiter plates was also used in this study. The results demonstrated that more bacteria adhered to the inner surface of stents perfused with bile containing 5 mg/mL mucin than of those perfused with bile containing 0.5 and 0 mg/mL mucin. Increased bacterial adherence was demonstrated on the stent surfaces conditioned with purified mucin compared with that seen on the nonconditioned stent surfaces. The optical densities indicating bacterial adhesion in the microtiter plate wells precoated with mucin were higher than those in non-coated plate wells. The in vitro results indicate that when a biliary stent is implanted in vivo, mucin in bile may condition the stent inner surface, modulate subsequent bacterial adherence to the surface, and participate in stent occlusion. PMID- 11873243 TI - Treatment of hemophilia B in mice with nonautologous somatic gene therapeutics. AB - The implantation of nonautologous cells encapsulated in immunoprotective microcapsules provides an alternative nonviral method for gene therapy. This strategy was successful in reversing the disease phenotypes of dwarfism and a lysosomal storage disease, mucopolysaccharidosis VII, in murine models. In this article we implanted transgenic hemophilic B mice with microcapsules enclosing factor IX-secreting C2C12 myoblasts to study the clinical potential of this approach in the treatment of hemophilia. Treated mice showed increased plasma factor IX levels as high as 28 ng of human factor IX per milliliter of plasma and decreased activated thromboplastin times (reduced by 20% to 29%). However, the level of factor IX decreased to baseline levels by day 7, coinciding with emergence of anti-human factor IX antibody, the titer of which increased greater than 10-fold by day 28. Monoclonal anti-CD4 antibodies were used to deplete CD4+ T cells to suppress the immune response against the recombinant factor IX. In the treated hemophilic mice, the anti-factor IX antibody response was totally suppressed to beyond day 28 accompanied by a significant decrease in activated thromboplastin time compared with that seen in untreated hemophilic mice. When the microcapsules were recovered from the intraperitoneal cavity after 38 days of implantation, the encapsulated cells continued to secrete factor IX at preimplantation levels, but both cell viability and microcapsule mechanical stability were reduced. Hence although the polymer chemistry of the microcapsules and cell viability may need to be improved for long-term delivery, nonautologous gene therapy with microencapsulated cells has been shown to be effective, at least for the short-term, in alleviating the hemophilic hemostatic anomaly. Coadministration of an immunosuppressant is effective in inhibiting antibody development against the delivered factor IX and should be considered for recipients at risk of inhibitor development. PMID- 11873244 TI - Potentiation of human lung fibroblast chemotaxis by the thromboxane A(2) analog U 46619. AB - Fibroblast production of extracellular matrix is crucial not only for normal tissue development and maintenance of tissue structure but also for the repair and remodeling processes after injury. Thromboxane A(2) (TXA(2)) is a potent mediator in inflammatory processes. The aim of this study was to investigate the effect of TXA(2) on chemotaxis of human fetal lung (HFL-1) fibroblasts induced by human plasma fibronectin or platelet-derived growth factor BB (PDGF-BB). By using the blindwell chamber technique, the TXA(2) agonist U-46619 alone had no chemotactic activity. However, U-46619 (200 nmol/L) stimulated HFL-1 fibroblast chemotaxis to human plasma fibronectin (20 microg/mL; 161.8% +/- 13.4%; P <.005) and to PDGF-BB (10 ng/mL; 188.5% +/- 7.0%; P <.005). Checkerboard analysis of human plasma fibronectin-directed migration confirmed that the TXA(2) agonist increased both chemotaxis and chemokinesis. The stimulatory effect of the TXA(2) agonist was concentration dependent and increased with time. Another agent known for stimulating the protein kinase C pathway, phorbol 12-myristate 13-acetate (10(-8) mol/L), had a similar effect, stimulating chemotaxis to fibronectin (146.2% +/- 8.6%). The stimulatory effect of the TXA(2) agonist on HFL-1 fibroblast chemotaxis was inhibited by the synthetic thromboxane receptor antagonist SQ29,548 (10(-5) mol/L) and the protein kinase C inhibitor calphostin (10(-7) mol/L). In summary, TXA(2) appears to stimulate fibroblast chemotaxis to fibronectin and PDGF, perhaps by modulating the rate of fibroblast migration. Such an effect may contribute to regulation of wound healing and the development of fibrotic disorders. PMID- 11873245 TI - The iron-loaded gerbil model revisited: effects of deferoxamine and deferiprone treatment. AB - Although the beneficial effects of deferoxamine (DFO) on iron-associated morbidity and mortality are well documented, the role of deferiprone (L1) in the management of transfusional iron overload is controversial. This debate involves not only the question of efficacy but also of safety, with particular emphasis on the risk of a paradoxical aggravation of iron toxicity by L1. We used the iron loaded gerbil model introduced by Carthew et al to compare the chelating efficacy of L1, DFO, or both in two gerbil strains treated by means of weekly iron-dextran injections: Psammomys obesus and pathogen-free Mongolian gerbils (Meriones unguiculatus). The difference between the high mortality and advanced hepatocellular necrosis observed in iron-loaded P obesus and the absence of mortality and limited morbidity encountered in pathogen-free Mongolian gerbils is most likely explained by the prevention of coincidental laboratory infections in the latter group. Iron-chelating treatment in all experimental groups resulted in a significant decrease in hepatic iron concentrations and normalization of mitochondrial respiratory enzyme activities, with combined L1 and DFO treatment being the most efficient, followed, in decreasing order, by DFO and L1 as single drug treatments. Judged by tissue iron concentrations, mitochondrial enzyme activity, and hepatic histology, we could find no evidence of a paradoxical aggravation of iron toxicity by L1 in either of the two series of studies. Although these data appear to be reassuring, the present controversy related to the role of L1 in the development of hepatic cirrhosis should be eventually settled by clinical studies evaluating the effects of long-term iron-chelating treatment. PMID- 11873246 TI - Expression of CD44 variants in colorectal carcinoma quantified by real-time reverse transcriptase-polymerase chain reaction. AB - CD44 is a family of transmembrane glycoproteins that has been linked to carcinogenesis and metastasis. It serves as a major receptor for hyaluronate. The v3 isoform binds to growth factors through heparan sulfate side chains and targets these factors to their high-affinity signal transducing receptors. The purpose of this study was to analyze the expression of CD44 v3 and v4 in human colorectal carcinoma with real-time reverse transcriptase-polymerase chain reaction (RT-PCR). Our results show that 19 of 56 cases (33.9%) showed a greater than 2-fold increase in CD44 v3 expression in tumors as compared with matched normal mucosa, while 15 of 44 cases (34.1%) showed a greater than 2-fold increase in CD44 v4 expression. There was a marked variation in fold-differences of CD44 gene expression between tumor and normal samples (T/N ratios) among the tumors. This prompted us to correlate the T/N ratios of the tumors with clinicopathologic characteristics. Interestingly, overexpression of CD44 v3 mRNA was associated with the presence of vascular invasion (P <.05). Similarly, overexpression of CD44 v4 was significantly correlated to increased depth of invasion (P <.05). Results from the present study suggest that overexpression of CD44 v3 and v4 mRNA levels may be useful clinical markers for colorectal carcinoma invasiveness. PMID- 11873247 TI - Genetics and the general pediatrician: where do we belong in this exploding field of medicine? PMID- 11873252 TI - Difficult problems in thyroid surgery. PMID- 11873253 TI - Prenatal toxoplasmosis diagnosis from amniotic fluid by PCR. AB - Toxoplasmosis is one of the most common infections all over the world. Most cases are asymptomatic, except in immunosuppressed individuals and fetuses, which can be seriously damaged. Prenatal diagnosis should be made as soon as possible since treatment of the mother can minimize fetal sequelae. Our aim in this study was to test the polymerase chain reaction technique (PCR) in 86 samples of amniotic fluid from women who seroconverted during pregnancy. DNA was amplified using external primers and, in a second step, internal primers, in a nested PCR system. Samples were also inoculated into mice and the newborn were evaluated by T. gondii serology, skull x-ray, transfontanel ultrasound, fundoscopic examination, lumbar puncture and clinical examination. PCR was positive in seven cases and negative in 79. Among PCR-positive cases, two were negative by inoculation into mice and by clinical evaluation; among PCR-negative ones, three had clinical evidence of toxoplasmosis and one was positive after inoculation into mice. PCR showed values of sensitivity = 62.5% and specificity = 97.4%; the values of inoculation into mice where 42.9% and 100%, respectively. Although PCR should not be used alone for prenatal diagnosis of congenital toxoplasmosis, it is a promising method and deserves more studies to improve its efficacy. PMID- 11873254 TI - Evaluation of IFN-gamma and TNF-alpha as immunological markers of clinical outcome in cutaneous leishmaniasis. AB - To evaluate if IFN-gamma and TNF-alpha levels could be used as markers of therapeutic response in cutaneous leishmaniasis, 54 patients with history of one ulcerated cutaneous lesion, with up to 30 days onset, were enrolled in the study. IFN-gamma and TNF-alpha were measured by ELISA in lymphocyte cultures supernatant before and 60 days after initiating therapy. Cure was considered to be a complete healing of lesion 60 days after treatment. IFN-gamma and TNF-alpha levels were similar in both groups of patients before therapy. There was a tendency to increase IFN-gamma levels in patients that were cured in 60 days, however the values did not reach statistical significance. In both groups of patients, TNF alpha levels were similar before therapy and fell significantly after treatment, irrespective of cure or maintenance of active lesion. PMID- 11873255 TI - The effects of immunization with recombinant Sm14 (rSm14) in reducing worm burden and mortality of mice infected with Schistosoma mansoni. AB - To investigate whether mice immunization with the recombinant form of a 14.7 KDa Schistosoma mansoni protein (rSm14) confers protection against a S. mansoni lethal challenge infection, rSm14-immunized mice were challenged with different cercarial burdens. A significant protection was detected in immunized mice challenged with 100 or 1,000 S. mansoni cercariae when compared with their controls (p< 0.004 and p< 0.01 respectively). Differently from previous report, none of the mice from the control group (not immunized and infected with 1000 cercariae) died before the 30th day post-infection. A direct correlation between the number of challenge cercariae and the precocity of mice death was found. IgM anti-rSm14 antibodies were significantly produced (p< 0.05) mainly in the groups of immunized mice infected with 500 or 1000 cercariae. IgG and IgA anti-rSm14 antibodies were not significantly detected. In Western immunoblots, all mice sera showed a specific antibody response with a 14.7 KDa antigen being reacted with particular intensity in sera from immunized mice. The results show that immunization with rSm14 reduced mice worm burden independently of the cercariae load of challenge infection. No correlation was found between serum antibodies and worm burden reduction. In relation to cercarial load and the rate and precocity of mice mortality a direct correlation was found. PMID- 11873256 TI - [Epidemiology and etiology of dermatophytosis in Goiania, GO, Brazil]. AB - Dermatophytes are a group of closely related fungi that have the capacity to invade keratinized tissue (skin, hair, and nails) of humans and animals to produce infections called dermatophytosis. In order to review the etiology and epidemiology of dermatophytosis in Goiania, GO, 1955 specimens with diagnostic suspicion of dermatophytic lesions, were collected from January to December, 1999, from the Mycology Laboratory in the Institute of Tropical Pathology and Public Health, Federal University of Goias. A total of 445 (22.8%) samples were positive for dermatophytes and Trichophyton rubrum was the most frequently isolated species (49.4%) followed by Trichophyton mentagrophytes (30.8%) and Microsporum canis (12.6%). Concerning the location of the lesions, the inferior limbs, feet and nails together with tinea capitis were the most frequently found clinical pattern in the majority of patients. Correlation between sex, age, location of the lesions and etiologic agents is considered in the study. PMID- 11873257 TI - [Epidemic dermatits by Paederus irritans in Piura, Peru at 1999, related to El Nino phenomenon]. AB - The phenomen El Nino that affected Peru at 1998, made possible the growth of copious vegetation in traditionally dry places. On this way, Paederus irritans, dipterous of the order Coleoptera that produces dermatitis when croushed on the skin, found substract that allowed the natural growth of its population. The coming of occasional rains on the months of February to April 1999, attacked the habitat of this insect, that looked for urban areas, exposing seriously the general population of Piura on north Peru to the contact. During February and May 1,451 cases of dermatitis caused by Paederus irritans, called latigazo because of its linear and eritematous characteristic lesions on the skin, were notified at Piura. The more affected body areas were head (56.6%) and neck (30.9%), but there were also cases of conjunctivitis and genital injuries, because of hands contamination. PMID- 11873258 TI - [Correlation among the positivity of the artificial xenodiagnosis and the amount of blood and triatomines used in the exam, in chronic chagasic patients]. AB - The aim of this study was to verify whether the amount of blood and number of triotomines used could improve the artificial xenodiagnosis performed in 200 chronic phase infected individuals. Ten or 40ml of peripheral blood was collected in heparinized (20.4 IU) vacuum tubes, and fed to 60 and 360 triatomines, respectively. Dipetalogaster maximus (1st instar, about 15 days after eclosion), as well as 3rd instar Triatoma infestans and Triatoma vitticeps and the 4th instar of Rhodnius neglectus were used. The faecal examinations were performed 30 and 60 days after xenodiagnosis procedure. The positivity with 10ml of blood was 19% and with 40ml, 44%, from which it was concluded that a correlation existed between the amount of blood and the number of triatomines used (p < 0.01). The positivity of the xenodiagnosis ranged from 9.7 to 100%, higher in young and adults up to 34 years old, but independent in relation to the sex of the chronic chagasic individuals studied. PMID- 11873259 TI - [Schistosoma mansoni: the action of lovastatin on the murine model]. AB - Aimed to evaluate the lovastatin action on Schistosoma mansoni oviposition, infected mice with 100 plus minus 10 cercariae of the LE strain were used. Thirty days after infection the animals were treated with 100, 200 and 400mg/kg of lovastatin, per os, during five consecutive days and then sacrificed 7, 15, 30 or 60 days after treatment. We analyzed: distribution of worms in mesenteries and liver; mortality of worms in the liver; alteration of the oogram; eggs counting in the jejunum and liver; presence of intrauterine eggs and morphology of the worms from the treated and control groups (infected and not treated animals). Significant statistical differences were found between treated and control groups when the presence of intrauterine eggs was considered and also, alteration on the oogram, eggs at different stages of development in jejunum, liver and in the body length of males and females. The morphological study of the worms showed that the degenerative modifications occurred, mainly in the reproductive system, with reduction and alteration of the viteline follicles and the ovary of the females. Also modifications in the males testicles were observed. The results suggest that the drug under study reduces, considerably, the female S. mansoni, oviposition increases the worms' size, leads to alteration in the reproductive system of males and females. It also may induce death of a significant part of the worm populations at the dose of 400mg/kg. PMID- 11873260 TI - [Microflora in chagasic megaesophagus]. AB - Chagasic patients with megaesophagus were submitted to an endoscopy of the upper digestive tract and the samples were collected with special instruments under sterilized conditions. One of the four samples collected was from the stase liquid and the other three samples were collected from fragments of the esophageal mucosa at one, three and five centimeters from the esophageal-stomach transition (Z line). The samples were analyzed by the Microbiology and Pathologic labs for the identification of microorganisms. After that, the results were correlated with the degree of mega esophagus according to Ferreira-Santos. We observed that the incidence of pathogenic microorganism is very high in megaesophagus, with no relation with the degree of dilatation making the surgery for the treatment of this affection potentially contaminated. There was no significant difference concerning the positivity of the culture relating to the degree of esophagus dilatation. PMID- 11873261 TI - [Recommendations for the treatment of bancroftian filariasis in symptomless and diseased patients]. AB - The goals of treatment for lymphatic filariasis are: to prevent, reverse, or halt progression of disease; and to interrupt transmission of the parasite. Selecting the appropriate therapy for the patient with lymphatic filariasis requires knowledge of the various clinical features of filarial disease and their pathogenesis. In the past, treatment of lymphatic filariasis has focused primarily on antiparasitic chemotherapy; however, for many of the acute and chronic manifestations of lymphatic filariasis, it is now clear that supportive or other forms of clinical care are even more important than antiparasitic medication in order to prevent worsening of the disease. Regardless of the clinical manifestations of filarial disease in a particular patient, the following three components of treatment should, in general, be considered: supportive or disease specific clinical care (including hygiene and diet), patient education and counseling and finally, antiparasitic chemotherapy with diethylcarbamazine (DEC) and/or the combination of DEC with ivermectin. The authors also describe the proportional efficacy of diethylcarbamazine and ivermectin, alone or in combination, for use in mass treatment aiming at transmission interruption and the use of hygiene as a public health approach for lymphedema prevention. PMID- 11873262 TI - [Tuberculosis: the negleted calamity]. AB - Epidemiological data on tuberculosis and the fight against the disease in Brazil and in the world are reviewed herein. Historical aspects of the different approaches to the control of the disease in Brazil are highlighted and the ongoing Brazilian Program for the Control of Tuberculosis is examined. To conclude, the author takes a look at unresolved problems and considers new alternatives for the control of tuberculosis. PMID- 11873263 TI - Paradoxical reaction to the treatment of tuberculosis uncovering previously silent meningeal disease. AB - The development of paradoxical clinical worsening following initiation of tuberculosis treatment may complicate the clinical course of both HIV-infected and uninfected patients. We report a severe manifestation of the so called paradoxical reaction to the treatment of tuberculosis that unmasked previously silent meningeal disease in a 34-year-old HIV-infected male patient. PMID- 11873265 TI - [Immunoblot as a supplemental test to detect antibodies to hepatitis C virus in blood donors]. AB - Supplemental tests using Immunoblot are recommended to improve specificity of anti-HCV by ELISA. In Brazil immunoblot is not officially recommended. Aiming to identify EIA false-positive rate 70 positive EIA anti-HCV blood donors were submitted to 3rd generation immunoblot at Hemocentro of Mato Grosso State where polymerase chain reaction tests are not performed. There were 44 (62.9%) immunoblot-positive, 22 (31.4%) negative and 4 (5.7%) indeterminate. Anti-HCV immunoblot can distinguish blood donors with false-positive ELISA from those who need medical assessment. Our data suggest that immunoblot could be useful in Brazilian blood banks where molecular biology tests are not available. PMID- 11873264 TI - Association of chagasic megacolon and cancer of the colon: case report and review of the literature. AB - There are few descriptions of association between chagasic megacolon and colon cancer. We report a case of obstructive abdomen caused by adenocarcinoma of the left colon in chagasic megacolon. A review of the literature revealed 8 cases of this association and, analyzing together the series of findings of cancer in chagasic organomegalies, we found a frequency of 4.8% in megaesophagus and 0.1% in megacolon. PMID- 11873306 TI - [Endometrial carcinoma - molecular pathogenesis meets histomorphology and treatment]. PMID- 11873266 TI - [Paradoxical reactions in the treatment of tuberculosis]. PMID- 11873307 TI - [Precancerous lesion of the endometrium and endometrial morphology in patients with tamoxifen therapy]. AB - The endometrioid type of endometrial adenocarcinoma,(type 1-carcinoma) is estrogen-dependent and frequently associated with endometrial hyperplasia. The nomenclature of these hyperplasias is currently under discussion. The highest risk for metachronous carcinoma is associated with atypical hyperplasia of the endometrium as detected in fractional curettings. In postmenopausal patients treatment should consist of abdominal hysterectomy. The so-called type 2 carcinomas, serous-papillary and clear-cell type, do not demonstrate a similar association with precursor lesions. Pathological findings in patients treated with Tamoxifen include endometrial atrophy and fibro-cystic endometrial polyps, sometimes with cellular metaplasias. Patients with breast cancer and tamoxifen treatment have an increased risk of endometrial carcinoma. In some of these patients it could be argued whether the carcinoma has developed in a proceeding endometrial hyperplasia. PMID- 11873308 TI - [Dualistic model of molecular pathogenesis in endometrial carcinoma]. AB - Sporadic endometrial carcinoma can be divided into two biologically and clinically distinctive subtypes of which one is estrogen-related (type I), the other estrogen-unrelated (type II). Type I carcinomas occur at younger age, express estrogen (ER) and progesterone receptors (PR), are frequently associated with endometrial hyperplasia and show a good prognosis. Type II carcinomas occur at older age, are negative for ER and PR, arise in the background of atrophic endometrium and show poor prognosis. Histologically, endometrioid carcinomas correspond to type I carcinomas whereas serous carcinoma is the prototype of type II carcinomas. Endometrioid and serous carcinomas are significantly different with respect to their molecular changes. Endometrioid carcinomas frequently show microsatellite instability (MIN), PTEN and K-ras mutation but infrequently p53 mutations, loss of p16 expression and her2/neu amplification, respectively. In contrast, serous carcinomas show a high frequency of p53 mutations and often loss of p16 expression whereas MIN and PTEN and K-ras mutations are uncommon. Familial endometrial carcinoma associated with HNPCC occur about two decades earlier than sporadic carcinomas, show endometrioid histology and are frequently MIN positiv due to germline mutations of mismatch repair genes (mostly MLH1 and MSH2). During the progression of endometrioid carcinoma PTEN mutations and MIN are considered early changes since they are present in a high frequency in atypical endometrial hyperplasia whereas p53 mutations, loss of p16 expression and her2/neu amplification are considered late events since they are predominantly found in poorly differentiated tumors. In contrast, p53 mutations are considered an early event in the pathogenesis of serous carcinoma occurring already in its putative precursor endometrial intraepithelial carcinoma (EIC). The future research will focus, besides the discovery of new relevant genes, on the interaction of known genes as well as their clinical relevance. PMID- 11873309 TI - [Role of estrogen receptor isoforms in the pathogenesis and treatment of endometrial cancer]. AB - Estrogens play a crucial role in the regulation of the physiology of breast tissue and endometrium. Furthermore, estrogen has been implicated in the initiation and progression of neoplasms of these tissues. Estrogens mediate their effects through various estrogen receptor isoforms and isotypes. In breast tissue and in the endometrium the classical estrogen receptor ERalpha represents the mainly expressed ER isoform, whereas in the ovary the alternative estrogen receptor ERbeta is predominantly expressed. This review briefly describes the structure, function and expression of these receptors with special regard to endometrial cancer. Recent data indicate that alterations of the physiological ERalpha/ERbeta ratio in endometrial cancer correlates with poor clinical outcome. The potential clinical relevance of differential ER-isotype expression is also discussed with respect to an antihormonal therapy. PMID- 11873310 TI - [Sonography and hysteroscopy for endometrial carcinoma and its precursors]. AB - The transvaginal sonography is a non invasive diagnostic method to evaluate the endometrium. It has a high reliability in the diagnosis of endometrial carcinoma, in the assessment of the depth of myometrial invasion as well as in preoperative staging. Patients with an episode of postmenopausal vaginal bleeding and an endometrial thickness (double layer) less-than-or-equal 4 mm should be controlled by transvaginal ultrasound examination after 3 month. In case of a endometrial thickness > 4 mm or in case of a persistent bleeding a histologic assessment should be obtained. Because of the highest sensitivity and specificity a hysteroscopic biopsy or a diagnostic hysteroscopy with subsequent D & C should be performed. Vaginal bleeding under hormonal replacement therapy should not be regarded to have more importance than vaginal bleeding in patients without HRT. After the current consensus hysteroscopy causes only a slightly increase in the rate of positive peritoneal cytology. This phenomenon seems to be of no influence on the disease free survival rate. PMID- 11873311 TI - [Cross-sectional imaging of endometrial carcinoma: MR imaging and CT]. AB - Magnetic resonance (MR) imaging has superseded computed tomography (CT) in the diagnostic assessment of cancer of the uterus (endometrial and cervical carcinoma). Contrast-enhanced MR imaging is highly accurate in staging endometrial carcinoma, in particular in terms of the depth of myometrial invasion. MR imaging will in the future provide a good basis for preoperative therapeutic decision making, especially to answer the question whether or not lymphadenectomy is necessary based on myometrial infiltration depth. - This article outlines the potential of MR imaging for staging endometrial cancer, provides a brief description of the imaging strategies, and discusses the role of MR imaging in the diagnostic assessment of endometrial cancer. PMID- 11873312 TI - [Treatment of endometrial carcinoma - evidence based criteria]. AB - The evaluation of decisions in treating endometrial cancer according to levels of evidence and grades for recommendation is gaining importance. This can help to evaluate results of studies and publications for guidelines and standards in treating endometrial cancer. - Concerning endometrial cancer there are some publications with a high level of evidence especially in early stages whereas data of treating progressive and advanced disease with a high level of evidence is lacking. - There are some studies with an acceptable level of evidence evaluating radiation and surgical treatment in stages I and II. In stages III and IV further data concerning evaluation of hormonal treatment and chemotherapy is pending. PMID- 11873313 TI - [Recommendations for the oncologic pathology report and morphologic factors associated with prognosis in endometrial carcinoma]. AB - Summary. The endometrial carcinoma (EC) is the fourth common malignant tumor in women with a reported incidence for Germany of 24.5. The prognostic evaluation shows an overall five-year survival of more than 76 %. The histopathologic report should include the tumor type in detail with special consideration of serous papillary, clear cell and endometrioid type. Stage T2 a and T2 b should only be diagnosed with clear-cut invasion of endocervical glands or the cervical stroma. In most cases of positive endocervical curetting, endmotrial carcinoma has been dislocated during fractional curetting and represents no indication for advanced surgery (Wertheim-Meigs-procedure). Special reference should be made on isthmic carcinoma, because there is a combination of lymphatic drainage of the cervix and corpus uteri. Well established prognostic factors are surgical tumor stage (pTNM system), pelvic lymph node metastases and tumor grading. Molecular biologic and cellkinetic parameters should be used for research purpose only. PMID- 11873315 TI - [Radiotherapy for endometrial cancer]. AB - Over the past decades, incidence, therapy and overall survival of endometrial cancer have changed considerably. Since 1977, surgical treatment has been preferred. In 1988, for example, only 5 % of the patients, mainly inoperable, were treated by primary irradiation. Patients, who undergo surgical treatment alone, have a significantly better outcome than patients with surgery and adjuvant irradiation. Patients treated by primary irradiation show the worst survival outcome, because of the selection bias for advanced stages. This observation even holds true, if adjusted for age and therapy modality. Modern irradiation technology may offer curative treatment options even in cases with advanced stages or severe co-morbidity, as well as locoregional recurrence. PMID- 11873314 TI - [Hormonal therapy and chemotherapy of endometrial cancer]. AB - Endometrial cancer is usually diagnosed at an early stage where surgery alone is the adequate therapy. Chemotherapy and hormonal treatment are therefore almost exclusively performed in palliative situations. Hormonal treatment with progestogens (medroxyprogesterone acetate and megestrol acetate) should be the therapy of choice primarily as these drugs are very well tolerated. Tamoxifen and GnRH analogs are further options but are seldom used. The response rates to hormonal treatment are relatively low (max. 25 %) with short remissions in most cases. - So far neither hormonal treatment nor cytotoxic chemotherapy has been shown to have substantial benefits in the adjuvant setting. In some selected high risk cases (serous papillary carcinomas, extra uterine manifestation) adjuvant chemotherapy may be an option following surgery, before or after radiotherapy. Age, general condition and morbidity of the patients need to be considered as limiting factors for chemotherapy. Crucial for the prognosis of all endometrial cancer patients however, is the stage adapted surgery. - Cytotoxic chemotherapy has failed to bring a break through in the therapy of advanced endometrial cancer. Cisplatin plus doxorubicin is the standard combination to date, with anthracyclines being the more important component. In a mono-therapy setting, doxorubicin and epirubicin are well tolerated and convenient in their efficacy. For recurrent and metastatic disease, docetaxel is being evaluated for efficacy and side effects in a multicenter phase II trial. PMID- 11873316 TI - [Recommendations for diagnosis and treatment in patients with endometrial carcinoma]. PMID- 11873317 TI - [Certified study protocols for patients with endometrial carcinoma]. PMID- 11873318 TI - [Psychosocial aftercare of patients with endometrial or cervical cancer]. AB - INTRODUCTION: Research in psychooncology focuses primarily on the impact of cancer, less on psychosocial causes of cancer. Recent tendencies in clinical practice are shown. PSYCHOSOCIAL PROBLEMS: Original papers show that anxiety, depression, postmenopausal symptoms, fatigue, sexuell issues, relationship and family become important problem areas for the patients. Anxiety and depression are independent of the state of the disease but, instead, depend on the extension of treatment. For unmarried single women sexual issues become a problem earlier whereas the problem develops later for married women. Women with cervical carcinoma are more affected than others. INTERVENTION: There is a high need for psychosocial counselling which is not considered enough in the daily clinical practice. Psychosocial aftercare is, regardless to different focuses, the task of the entire team. Good precare is just as important as good aftercare to reduce anxiety, pain and increase well-being. Addressing information and support in a timely fashion is important in providing success to psychosocial aftercare. PMID- 11873319 TI - [Psychooncological aspects in the care of females in HNPCC families]. AB - HNPCC families are characterized by a genetic predisposition for colorectal and other cancers. In Dusseldorf we pursue an interdisciplinary counselling approach (geneticists, psychologists and surgeons). Apart from the genetic counselling itself, special emphasis is placed on the benefits of surveillance and screening recommendations. In this pilot study we were interested in investigating if the counselled women had understood the increased risk of gynecological cancers and correspondingly underwent the intensified screening modalities. We selected the approach of a telefone interview and encountered broad acceptance for contacting persons this way. Affected women are better compliant to screening recommendations than women at risk. Both groups consider colonoscopy more strainful than gynecological surveillance. A regular transvaginal ultrasound is not performed in some 40 % of at risk women although 2/3 of them see their gynecologist in regular intervals. Given the support of gynecologists a higher rate of transvaginal ultrasound in women at risk should be achievable. PMID- 11873321 TI - Person-environment mutual process: studying and facilitating healthy environments from a nursing science perspective. PMID- 11873320 TI - A PEX6-defective peroxisomal biogenesis disorder with severe phenotype in an infant, versus mild phenotype resembling Usher syndrome in the affected parents. AB - Sensorineural deafness and retinitis pigmentosa (RP) are the hallmarks of Usher syndrome (USH) but are also prominent features in peroxisomal biogenesis defects (PBDs); both are autosomal recessively inherited. The firstborn son of unrelated parents, who both had sensorineural deafness and RP diagnosed as USH, presented with sensorineural deafness, RP, dysmorphism, developmental delay, hepatomegaly, and hypsarrhythmia and died at age 17 mo. The infant was shown to have a PBD, on the basis of elevated plasma levels of very-long- and branched-chain fatty acids (VLCFAs and BCFAs), deficiency of multiple peroxisomal functions in fibroblasts, and complete absence of peroxisomes in fibroblasts and liver. Surprisingly, both parents had elevated plasma levels of VLCFAs and BCFAs. Fibroblast studies confirmed that both parents had a PBD. The parents' milder phenotypes correlated with relatively mild peroxisomal biochemical dysfunction and with catalase immunofluorescence microscopy demonstrating mosaicism and temperature sensitivity in fibroblasts. The infant and both of his parents belonged to complementation group C. PEX6 gene sequencing revealed mutations on both alleles, in the infant and in his parents. This unique family is the first report of a PBD with which the parents are themselves affected individuals rather than asymptomatic carriers. Because of considerable overlap between USH and milder PBD phenotypes, individuals suspected to have USH should be screened for peroxisomal dysfunction. PMID- 11873322 TI - Policy, procedure, and routine: matters of moral influence. PMID- 11873323 TI - Becoming invisible: elder as teacher. PMID- 11873324 TI - Who I am. PMID- 11873325 TI - Human becoming practice with children. PMID- 11873326 TI - The nurse theorists: 21st-century updates--Rosemarie Rizzo Parse. PMID- 11873327 TI - Energy: a bridging concept for nursing science. AB - A philosophical inquiry into the concept of energy revealed two ideas of energy across multiple disciplines. Analysis of the conceptual models of Nightingale, Levine, and Rogers supported the presence of two paradigmatic views of energy in nursing science that, while divergent, share a common theme. The outcome of this inquiry leads to the tentative belief that there is unity in diversity and may perhaps lead to refinement of existing nursing theory and a more congruent framework for scientific inquiry. This article specifically addresses the nursing theoretical implications related to ideas of energy arising from the inquiry. PMID- 11873328 TI - Adaptation in children with cancer: research with Roy's model. AB - This qualitative study was undertaken to establish a conceptual framework for the adaptation process of Taiwanese children with cancer. It was guided by Roy's adaptation model. Thirty-four pediatric cancer patients (23 boys and 11 girls) with different illness trajectories (from newly diagnosed to terminal stage) participated. Data were collected through individual in-depth interviews with patients and primary caregivers, observations, medical chart review, and researchers' reflective journals. The following five aspects of adaptation that emerged from the study were categorized according to different age groups: physiological and psychological, cognitive (self-concept), interdependence, and future orientation (self-concept and role). Comparison with Roy's adaptation theory is discussed. PMID- 11873331 TI - Nursing theories and their relation to knowledge development in Iceland. PMID- 11873330 TI - Borrowed theories, shared theories, and the advancement of nursing knowledge. AB - Despite the continued use of borrowed theories in nursing, little attention has been given to determining whether theories developed in another discipline are empirically adequate descriptions, explanations, or predictions of nursing phenomena. In this article, we demonstrate how a borrowed theory can be placed within a nursing context by linking it with two different conceptual models of nursing. We present our plans for research focused on condom use behavior and discuss how results from these studies will be used to determine whether the borrowed theory can be considered a shared theory. PMID- 11873329 TI - Empirical tests of the Neuman systems model: relational statement analysis. AB - To be a valid test of the Neuman systems model, researchers' relational statements must be congruent with Neuman's axioms and must also be congruent at the conceptual, theoretical, and operational levels. Guided by Cooper's five stage integrative review method, 92 quantitative research studies (dissertations and journal articles) were collected and categorized according to an expanded version of Silva's levels of theory testing. Nine studies, with explicit relational statements, were analyzed at three levels of abstraction, and interpretations of the studies' results were explored. Recommendations are made for the establishment of a program of research. PMID- 11873332 TI - Nursing theory-guided practice: what it is and what it is not. PMID- 11873333 TI - The universe is flat. PMID- 11873334 TI - Bearing witness-not bearing witness as synergistic individual-community becoming. PMID- 11873335 TI - Further reflections on energy fields. PMID- 11873336 TI - Nursing research on the health patterning modalities of therapeutic touch and imagery. PMID- 11873337 TI - Advance directives: living with certainty-uncertainty--a nursing perspective. PMID- 11873338 TI - Reflections on suffering. PMID- 11873339 TI - Helping students to know and respond to human suffering. PMID- 11873341 TI - The nurse theorists: 21st-century updates--Betty Neuman. Interview by Jacqueline Fawcett. PMID- 11873340 TI - Prescription, freedom, and participation: drilling down into theory-based nursing practice. PMID- 11873342 TI - Adaptation as a mediator of intimate abuse and traumatic stress in battered women. AB - The purpose of this research was to test one aspect of a proposed middle-range theory synthesized from the Roy adaptation model and linked with the framework of post-traumatic stress. A predictive-correlational design was used to examine adaptation as a mediator of traumatic stress in battered women. Analysis included regression and path analytic procedures. Results indicated (a) direct relationships between the focal stimuli of abuse and the response of post traumatic stress disorder and (b) adaptation in the physiologic, self-concept, and interdependence modes partially mediated the relationship between the focal stimuli of abuse and the response of post-traumatic stress disorder in battered women. PMID- 11873343 TI - Feeling understood: a melody of human becoming. AB - This phenomenological-hermeneutic study centered on the phenomenon of feeling understood, which was conceptualized by the researcher as a melody of human becoming significant to quality of life. For the first time the Parse research method was used with music as part of the dialogical engagement. The study was conducted with 10 women living with an enduring health situation who volunteered to be in tape-recorded dialogue with the researcher to discuss feeling understood and to create a musical expression of this phenomenon. The finding of this study, which is the structure of the lived experience of feeling understood, surfaced from the dialogues and musical expressions: Feeling understood is an unburdening quietude with triumphant bliss arising with the attentive reverence of nurturing engagements, while fortifying integrity emerges amid potential disregard. PMID- 11873344 TI - Neuman systems model-based research: an integrative review project. AB - The project integrated Neuman systems model-based research literature. Two hundred published studies were located. This article is limited to the 59 full journal articles and 3 book chapters identified. A total of 37% focused on prevention interventions; 21% on perception of stressors; and 10% on stressor reactions. Only 50% of the reports explicitly linked the model with the study variables, and 61% did not include conclusions regarding model utility or credibility. No programs of research were identified. Academic courses and continuing education workshops are needed to help researchers design programs of Neuman systems model-based research and better explicate linkages between the model and the research. PMID- 11873345 TI - The spiritual care meanings of adults residing in the midwest. AB - Only limited nursing knowledge exists as theoretical guidance for nurses in providing spiritual care. Using Leininger's theory of culture care diversity and universality, the purpose of this ethnonursing research study was to discover the embedded spiritual care meanings, expressions, lived experiences, and practices of adults residing in the Midwest and their perceptions of spiritual nursing care. Data were collected through interviews of 6 key and 12 general informants. Five universal spiritual themes were supported by the findings. Culture care modes were used to explicate spiritual knowledge that can be integrated into nursing practice. PMID- 11873346 TI - Caring narratives and the strategy of presence: narrative communication in nursing practice and research. AB - Professional nursing care is formed and carried out in a social cultural process. The discipline of nursing should study narrative communication to understand how individual and collective levels are connected in experiences of sickness and cure. Narrative as transformative acts of caring and the narrative structure of preverbal acts and contexts of care are waiting for further research and are naturally connected to the art of nursing. Personal presence, listening, and engagement are important to ethnographic research with a focus on narrative as well as to first-rate nursing care. PMID- 11873347 TI - The acceptance of nursing theory in Japan: a cultural perspective. PMID- 11873349 TI - Institutional ethics committees: a nursing perspective. PMID- 11873348 TI - Critical thinking: toward a nursing science perspective. AB - Critical thinking is an important phenomenon in nursing science because of its implications for education, practice, and the advancement of nursing knowledge. As a context-dependent, evolving life process, critical thinking appears to be congruent with assumptions and principles of SUHB. Thus, it may be asserted that critical thinking arises within the mutual process of human and environment and thus is a pattern manifestation of the human-environment field process. Before this assertion can be fully accepted, however, much investigation is needed. Nurse scholars are called upon to re-examine critical thinking and to consider possibilities that, until now, have been neglected or unimagined. To date, the positivistic view of critical thinking has yielded limited information. Nurse scientists, therefore, have an opportunity to extend and refine knowledge of critical thinking by embarking upon new and exciting avenues of discovery. PMID- 11873350 TI - An invitation to the unfamiliar: engaging the religion-science interface. PMID- 11873351 TI - Self-care deficit nursing theory and the nurse practitioner's practice in primary care settings. PMID- 11873352 TI - The nurse theorists: 21st-century updates--Dorothea E. Orem. PMID- 11873353 TI - A theory of dependent-care: a corollary theory to Orem's theory of self-care. AB - Dependent-care has its origins in people's requirements for regulatory care. The foundations for dependent-care are found in the ability of individuals to provide their required care. First introduced as a corollary to self-care, this work emerged through a process of reading and discussion. Models that support the theory of dependent-care are identified. Premises are stated. There is elaboration of the conceptualizations representing the work that has been done. There are still elements that need further development, such as specifying the enabling abilities of dependent-care agency and verifying and formalizing the various elements presented. PMID- 11873354 TI - Self-care: a foundational science. AB - Further development of conceptual elements of the theory of self-care, one of the three constituent theories of Orem's self-care deficit theory of nursing, is reported. Five content areas of a practical science of self-care are identified; one content area, self-care requisites, is refined and developed. The nature of self-care requisites is reformulated; guides and standards for the expression of self-care requisites, examples of expressed self-care requisites, and a self-care practice guide are described. These developments are illustrated using the example of the requisite to maintain an adequate fluid intake. PMID- 11873355 TI - The structure of self-care in a group of elderly people. AB - According to Orem's self-care deficit theory of nursing, the structure of self care consists of self-care agency balanced by therapeutic self-care demand. Different conditioning factors constitute these two constructs. The aim of this study was to investigate through secondary analysis the structure of self-care in a group of elderly. Data were originally collected from a total of 125 randomly chosen elderly individuals (65+ years of age) in Sweden by means of a mailed questionnaire. Confirmatory factor analysis was used to show that self-care agency was totally and significantly balanced against therapeutic self-care demand and explained by five conditioning factors. PMID- 11873356 TI - Understanding homeless adults by testing the theory of self-care. AB - This article explicates, within a sample of homeless adults, the relationship between self-care agency, self-care, and well-being, while controlling for select basic conditioning factors. Self-care is found to explain 30% of the variance in current well-being. Self-esteem and affect balance, along with the power components of self-care agency, are found to explain 25% of the variance in self care and, together with select basic conditioning factors, to explain 54% of its variance. Affect is proposed as an energy disposition, a dimension of self-care agency. Recommendations are made to strengthen the homeless individual's self care agency. PMID- 11873357 TI - The challenge of nursing in Guyana. PMID- 11873358 TI - Orem's general theory of nursing and families. PMID- 11873360 TI - A nursing theory of personal system empathy: interpreting a conceptualization of empathy in King's interacting systems. PMID- 11873359 TI - A nursing theory of personal system empathy: interpreting a conceptualization of empathy in King's interacting systems. PMID- 11873361 TI - Roy adaptation model and perspectives on the family. PMID- 11873362 TI - Language and the sow-reap rhythm. PMID- 11873363 TI - Interdisciplinarity and nursing: "everything is everything," or is it? PMID- 11873364 TI - Relational complexity: from grounded theory to instrument development and theoretical testing. PMID- 11873365 TI - The ethics of bearing witness in healthcare: a beginning exploration. PMID- 11873366 TI - On global health and justice: a nursing theory-guided perspective. PMID- 11873367 TI - Advancing a global perspective: the world as classroom. PMID- 11873368 TI - From silence to voice: knowledge, values, and beliefs guiding healthcare practices with persons living with dementia. PMID- 11873369 TI - The nurse theorists: 21st-century updates--Imogene M. King. Interview by Jacqueline Fawcett. PMID- 11873370 TI - An international human becoming hermeneutic study of Tom Hegg's A cup of Christmas Tea. AB - This article seeks to contribute to human becoming theory and to nursing by providing an international human becoming hermeneutic study of Thomas Hegg's A Cup of Christmas Tea. The human becoming hermeneutic method was used in this study to discover emergent meanings about human experiences. Guided by the method, the authors discovered three emergent meanings: honoring the cherished; communing with the was, is, and will be; and triumphing with new vision. These meanings were synthesized by the authors. A Cup of Christmas Tea is the story of the way triumphing with new vision arises with honoring the cherished in communing with the was, is, and will be. The conclusion for families and nurses is that by remaining open to all possibilities that exist in each now, moments of serendipitous togetherness can transform human trepidation and negative views of later life. PMID- 11873371 TI - Mexican American family survival, continuity, and growth: the parental perspective. AB - An ethnographic study using Roy's adaptation model was conducted among 23 Mexican American families in Hidalgo County, Texas, from 1994 to 1998. The purpose was to characterize the family goals of survival, continuity, and growth from the parental perspective during early family formation. Parents affirmed that being healthy, being a united couple, having supportive parents, having a steady job, and having civic harmony were essential characteristics of family survival. Family continuity was characterized by mothers doing tasks inside the house, father doing tasks outside the house, and both parents performing toddler and early childhood tasks. Family growth was characterized by having shared communication, growing in togetherness, planning ahead, exerting joint effort, and helping the child become part of the family. PMID- 11873372 TI - The lived experience of contentment: a study using the Parse research method. AB - The purpose of this study was to answer the research question. What is the structure of the lived experience of contentment? The participants were 10 women volunteers. The Parse research method, a phenomenological-hermeneutic method, was used to uncover the meaning of contentment. The major finding of this study is the structure: Contentment is a satisfying calmness amid the arduous as resolute liberty arises with benevolent engagements. The structure provides knowledge about contentment and its connection to health and quality of life. It is discussed in relation to the principles and concepts of human becoming and in relation to how it can inform future research and practice. PMID- 11873373 TI - Using conceptual models of nursing to guide nursing research: the case of the Neuman systems model. AB - Conceptual models of nursing inform thinking and give meaning and direction to nursing research. The Neuman systems model is used to exemplify the following five steps, which provide specific direction for conceptual model-based research: (a) Develop a comprehensive understanding of the substantive content and research rules of the conceptual model, (b) review existing research guided by the conceptual model, (c) construct a conceptual-theoretical-empirical structure, (d) clearly communicate the conceptual-theoretical-empirical structure, and (e) conclude the report with an evaluation of the empirical adequacy of the middle range theory and the credibility of the conceptual model. PMID- 11873374 TI - Developing perspectives on Korean nursing theory: the influences of Taoism. AB - Nursing theory provides a systematic explanation and description of nursing phenomena. Western nursing theories have widely influenced Korean nursing. And yet, although nursing theory has universal aspects, the differences in philosophy and culture that are unique to each country need to be considered. This inquiry seeks to investigate the Korean cultural heritage, which integrates Confucianism, Buddhism, and Taoism, and how it provides a unique worldview of human beings, the universe, health, and nursing. Essential principles and therapies consistent with Taoist philosophy are also identified. This framework is proffered as the basis for establishing understanding between Korean nurses and patients. PMID- 11873375 TI - Are nurses advancing nursing knowledge? PMID- 11873376 TI - Nursing knowledge in a mostly French-speaking Canadian province: from past to present. PMID- 11873377 TI - "The role of nursing theory in standards of practice: a Canadian perspective.". PMID- 11873378 TI - The value of economic modeling studies in the evaluation of treatment strategies for multiple sclerosis. PMID- 11873379 TI - Regulatory issues for health-related quality of life--PhRMA Health Outcomes Committee workshop, 1999. AB - INTRODUCTION: Health-related quality-of-life (HRQL) can be defined as the impact of disease and treatment across the physical, psychological, social and somatic domains of functioning and well-being. Health-related quality-of-life measures are included in clinical trials of drug treatment to assess the impact of therapy on the patient's functioning. HRQL guidance could allow for use of this data in drug labeling and promotion. OBJECTIVES: The aim of our study was to provide recommendations with respect to regulatory issues important to the development of guidelines for HRQL research. METHODS: The HRQL workshop was planned jointly by members of the Pharmaceutical Research and Manufacturers of America Health Outcomes Committee and the Division of Drug Marketing, Advertising, and Communications of the Food and Drug Administration. The workshop was limited to six regulatory issues related to HRQL research in clinical trials of pharmaceutical therapies. These six issues were: instrument selection and validation, study design, data analysis, HRQL and safety, clinical meaning, and promotional use. Before the meeting, a consensus was reached that HRQL does not measure, nor should it be used to measure, safety. Therefore, five work groups discussed HRQL issues and made recommendations. RESULTS: Overall, the workshop recommended that HRQL measures be treated as any other clinical end point. The workshop recognized that research in HRQL methods is ongoing and that any guidance should be flexible to allow for changes in this developing research area. CONCLUSIONS: HRQL provides a patient perspective on the impact of disease and therapy on patients' daily life and functioning. Including HRQL information in promotion could be beneficial to decision making on the use of therapies. HRQL is a measure of effectiveness, not safety, and should be treated as any other clinical end point. PMID- 11873380 TI - The direct cost and incidence of systemic fungal infections. AB - OBJECTIVES: In this study we determined the incidence and direct inpatient and outpatient costs of systemic fungal infections (candidiasis, aspergillosis, cryptococcosis, histoplasmosis) in 1998. METHODS: Using primarily the National Hospital Discharge Survey (NHDS) for incidence and the Maryland Hospital Discharge Data Set (MDHDDS) for costs, we surveyed four systemic fungal infections in patients who also had HIV/AIDS, neoplasia, transplant, and all other concomitant diagnoses. Using a case-control method, we compared the cases with controls (those without fungal infections with the same underlying comorbidity) to obtain the incremental hospitalization costs. We used the Student's t-test to determine significance of incremental hospital costs. We modeled outpatient costs on the basis of discharge status to calculate the total annual cost for systemic fungal infections in 1998. RESULTS: For 1998, the projected average incidence was 306 per million US population, with candidiasis accounting for 75% of cases. The estimated total direct cost was $2.6 billion and the average per-patient attributable cost was $31,200. The most commonly reported comorbid diagnoses with fungal infections (HIV/AIDS, neoplasms, transplants) accounted for only 45% of all infections. CONCLUSIONS: The cost burden is high for systemic fungal infections. Additional attention should be given to the 55% with fungal disease and other comorbid diagnoses. PMID- 11873381 TI - Outcomes research in the health-care system: driven by results. PMID- 11873382 TI - The incidence and cost of hospitalization for 5-FU toxicity among Medicare beneficiaries with metastatic colorectal cancer. AB - OBJECTIVE: To estimate the incidence and associated cost of hospitalizations for toxicities associated with 5-fluorouracil (5-FU) among patients with metastatic colorectal cancer. METHODS: Using the 1994 Medicare 5% sample, we identified all patients with metastatic colorectal cancer who had undergone colorectal surgery. We then stratified them into those who received 5-FU therapy within 90 days of their surgery (5-FU group) and those who did not receive chemotherapy (no chemotherapy group); patients who received chemotherapeutic agents other than 5 FU were excluded from the sample. Using techniques of survival analysis, we then compared the incidence and associated cost of all hospital admissions with listed International Classification of Disease, Ninth Revision, Clinical Modification (ICD-9-CM) diagnostic codes (primary or secondary) for conditions possibly related to 5-FU toxicity (e.g., volume depletion, stomatitis, nausea, and vomiting). RESULTS: A total of 441 patients met all study entry criteria, including 192 who received 5-FU and 249 who did not receive chemotherapy following surgery. Patients in the 5-FU group were younger than those in the no chemotherapy group (p < .001). Mean (+/- SD) follow-up time was slightly longer in the 5-FU group (137 +/- 96 days vs. 117 +/- 88 days for no chemotherapy). The incidence of toxicity-related hospitalizations at 10.5 months (principally volume depletion, agranulocytosis, gastroenteritis, and nausea and vomiting) was 31% among patients who received 5-FU and 8% among those who did not receive chemotherapy. The cost of inpatient care at 10.5 months was $2716 higher among 5 FU patients. CONCLUSIONS: Hospitalization for toxicity of Medicare patients with metastatic colorectal cancer receiving 5-FU is frequent and costly. PMID- 11873383 TI - Cost-effectiveness analysis of interferon beta in multiple sclerosis: a Markov process analysis. AB - OBJECTIVE: The objective of this study was to examine the cost-effectiveness of preventive treatment with interferon beta (IFNB) versus no preventive treatment in patients with multiple sclerosis. METHODS: The setting for this study was the United Kingdom. A lifetime Markov process model was constructed to model the average quality-adjusted life years (QALYs) and the costs of both treatment strategies. Data for the construction of the model came from published literature, including large multicenter randomized clinical trials in relapsing remitting and secondary progressive multiple sclerosis. Costs were obtained from published sources. RESULTS: The results of the baseline analysis from the National Health Service (NHS) perspective showed that the use of interferon beta as preventive treatment for MS increased the total average discounted cost from 51,214 Pounds to 221,436 Pounds per patient. The undiscounted effectiveness increased from 24.9 QALYs to 28.2 QALYs, resulting in an incremental cost effectiveness ratio of 51,582 Pounds per QALY. Sensitivity analyses showed the robustness of this model for other interferons. CONCLUSION: The study showed that preventive treatment with interferon beta in patients with multiple sclerosis may not be fully justified from a health-economic perspective, although interferon beta is associated with an improved effectiveness compared with no preventive treatment. PMID- 11873384 TI - The economics of gene therapy and of pharmacogenetics. AB - This paper focuses on the economic issues arising from two uses of genomics: 1) the development of gene therapy; 2) and use of pharmacogenetics to identify a patient's genotype before treatment to exclude those who will not benefit or who may be harmed. We conclude that private-sector investment aimed at developing gene therapy for monogenic diseases is likely to be socially suboptimal. Short term administration regimens yielding long-term therapeutic benefits are likely to meet payer resistance to large "one-off" costs because of budget constraints or, in competitive systems, concerns that the savings would accrue to future insurers or would attract high-cost patients. For some monogenic diseases, patient numbers may be too small to support commercial development without changes to orphan drug legislation or payer willingness to accept higher cost effectiveness thresholds. In the case of pharmacogenetics, we conclude that it can often be socially optimal to test before treatment, particularly if the proportion of nonresponders is high, if there is a potential for serious adverse reactions, or if the test is inexpensive. Genetic testing that fragments the patient population could reduce incentives for R&D unless prices are adjusted to reflect the higher expected benefits of targeted treatment per patient. Even in situations where prices are adjusted, patient populations may be too small to make commercial development viable. This problem with small numbers is analogous to that associated with gene therapy for monogenic diseases and may require similar remedies if society values developing treatments for these diseases. PMID- 11873385 TI - A method for identifying the financial burden of hospitalized infants on families. AB - OBJECTIVES: To describe a method for measuring the direct and indirect costs to families of infants hospitalized with respiratory syncytial virus (RSV). METHODS: After pretesting and revising a questionnaire, a prospective survey was conducted in multiple tertiary-care hospitals with pediatric intensive care units. Eligible patients were infants less than 12 months old who had not received RSV prophylaxis and were hospitalized with a confirmed RSV infection. All English- and Spanish-speaking caregivers of eligible subjects were asked to participate in a face-to-face, structured interview on the day of hospital discharge regarding hospitalization-related direct and indirect costs. Thirty days later, caregivers were re-interviewed by telephone about their RSV-related costs during the elapsed month. The survey was initiated in February 2000 and continued through April 2001. RESULTS: In addition to the infants' parents, numerous adults visited 55% of hospitalized infants. In 17% of cases, nonparents missed work to visit the child. Volunteers watched siblings of 26% of the infants. Relying only on closed ended questions about parents' costs during the hospitalization would have missed important information about child-care volunteers and types of expenses. Follow up interviews revealed that RSV-related out-of-pocket expenses and missed work continued during the month following discharge. CONCLUSIONS: Survey instruments should be pretested with potentially eligible subjects. Open-ended questions are needed, because all costs cannot be anticipated. Respondents should be probed for details. This method revealed certain time and financial burdens during and after hospitalization that had not been previously reported in the literature. PMID- 11873386 TI - Modeling for health care and other policy decisions: uses, roles, and validity. PMID- 11873387 TI - Modeling for health care and other policy decisions: uses, roles, and validity. PMID- 11873388 TI - Bioterrorism and biodefence. PMID- 11873389 TI - Stroke in elderly; identification of risk factors. AB - BACKGROUND: This study was undertaken to identify stroke risk factors prevalent in our elderly population. METHODS: The subjects included 100 consecutive stroke patients with recent stroke, at or above the age of fifty years, both sexes inclusive, who presented at medical units of Liaquat Medical College Hospital Hyderabad. A detailed history of the patients was taken, thorough clinical examination was done and various laboratory tests were carried out to identify all the possible risk factors for stroke in the subjects. RESULTS: Most important risk factors prevalent in our population were found to be hypertension (64%), diabetes mellitus (29%), smoking (29%), heart disease (23%), obesity (17%) and hypercholestraemia (15%). CONCLUSION: It is concluded that in elderly stroke patients, many risk factors are identified. Awareness of these risk factors, their early and effective treatment and adaptation of various preventive measures is warranted. PMID- 11873390 TI - Interlocked nailing of comminuted fractures shaft of femur. AB - BACKGROUND: Comminuted fracture shaft of femur are difficult to treat. Internal fixation with ordinary nail does not hold the fracture fragments while osteosynthesis with plates and screws does not always produce stable fixation and involves an increased risk of infection. METHODS: Thirty six comminuted fractures shaft of femur were treated with closed interlocked nailing. There were twenty closed and sixteen open fractures. RESULTS: Thirty six fractures united without additional surgical intervention. Average time to union was 34 weeks (range 26 to 64 weeks). In five patients secondary bone grafting was done. CONCLUSION: Closed Interlocked nailing is the treatment of choice for most comminuted fracture shaft of femur. PMID- 11873391 TI - Prevalence of thyroid microsomal and thyroglobulin autoantibodies in goitrous lesions. AB - BACKGROUND: This prospective study was done to evaluate the serum levels of Microsomal and Thyroglobulin autoantibodies in patients with toxic diffuse goiter and nodular goitre (non-toxic) undergoing radio-iodine therapy and thyroidectomy respectively. METHODS: Forty eight patients suffering from thyroid disorders, 29 with nodular goiter and 19 with toxic diffuse goitre (TDG) and 15 age & sex matched normal controls were studied. Thyroid microsomal (MSAb) and thyroglobulin autoantibodies (TGAb) were estimated in the sera of all the subjects using the commercially available kits based on tanned red cell haemaglutination technique. RESULTS: MSAb seropositivity in TDG and nodular goitre was found to be 78.9% and 51.7% respectively. On the other hand, TGAb seropositivity was 57.9% and 27.6% in cases of TDG and nodular goitre. 13.3% and 6.7% of the normal controls were positive for MSAb and TGAb respectively. CONCLUSION: These findings indicate that autoimmunity is implicated in the genesis of commonly occurring thyroid disorders. PMID- 11873392 TI - Metabolic effects of alcoholism and its relationship with alcoholic liver disease. AB - BACKGROUND: Chronic and excessive ethanol consumption is associated with cellular proliferation, fibrosis, cirrhosis and cancer of liver. It was planned to study the effect of alcohol on different biochemical parameters as distribution of these parameters may lead to several complications in gastrointestinal tract, liver, kidney, brain etc. METHODS: 50 alcoholic males and 20 normal subject (with no history of alcohol) with an age ranges 30-45 years were included in the study. Biochemical parameters related to liver were estimated by standard kit. Besides, ions and electrolytes were also determined by standard kit and flame photometry. RESULTS: It was observed that level of alkaline phosphatase, alanine transferase, protein and globulin were significantly increased as compared to normal subjects. Besides, ions like calcium and phosphate were significantly decreased. On the other hand the level of potassium and magnesium was significantly decreased as compared to normal subjects. Electrophoresis shows a protein of 100 Kda is present in the patients sample as compared to control subjects. CONCLUSION: It is therefore concluded that abnormal biochemical function of liver in alcoholism can lead to several complications, hence further research is needed to reach a definite conclusion. PMID- 11873393 TI - Diagnosis of deep vein thrombosis in the leg by using colour coded duplex sonography. AB - BACKGROUND: Deep Vein Thrombosis (DVT) is regarded rare [not readable: see text] Asia. There is no documentation of the incidence of DVT in Pakistan. Clinical diagnosis of Deep Vein Thrombosis is inaccurate. Venography is the most reliable method of diagnosis, but it has several disadvantages. Many non-invasive diagnostic methods have therefore been developed in the past decade. Of these only duplex sonography has comparable accuracy. METHODS: We studied 100 cases of suspected DVT by using colour coded duplex sonography. We report the results of a prospective study in patients with suspected deep vein thrombosis evaluated by both colour coded Doppler sonography and venography. CONCLUSION: We conclude that colour coded duplex sonography is a highly accurate, simple, non-invasive method for detecting femoropopliteal thrombosis. Additional venography is not necessary. Its value in diagnosing isolated calf vein thrombosis remains to be established. PMID- 11873394 TI - Effect of oral and injectable contraceptives on serum calcium, magnesium and phosphorus in women. AB - BACKGROUND: This study aims at evaluating serum calcium, magnesium and phosphorus levels in women taking oral and injectable contraceptives. METHODS: Serum calcium, magnesium and phosphorus were measured in 50 women taking oral contraceptives (Lofeminal) and 50 women taking injectable contraceptive (Depo medroxy progesterone acetate and Norigest). These women were used as controls before starting these contraceptives. RESULTS: There was significant decrease in serum levels of calcium, magnesium and phosphorus in women taking oral contraceptives but there was significant increase in these minerals in women taking injectable contraceptives. CONCLUSIONS: It is suggested that these contraceptives should be used with due care and with proper investigations of the women before and during the therapy. PMID- 11873395 TI - The optimum dose of nicotinamide for protection of pancreatic beta-cells against the cytotoxic effect of streptozotocin in albino rat. AB - BACKGROUND: The natural course of Insulin Dependent Diabetes Mellitus is characterized by progressive destruction of insulin producing beta-cells of the pancreas resulting from an autoimmune process. The toxic effect of some beta cells toxins like streptozotocin (used to produce animal models of IDDM) has been associated with the oxidative stress due to enhanced DNA repair and NAD depletion in damaged beta-cells. This activity of streptozotocin has been prevented with the use of nicotinamide. METHODS: A light microscopic study was designed to determine the optimum dose of nicotinamide required for protection of pancreatic beta-cells against the toxicity of streptozotocin. 35 adult male albino rats were divided into five equal groups A, B, C, D and E. the duration of study was 14 days. The animals in experimental groups C, D and E received a single intraperitoneal injection of nicotinamide 250 mg/Kg, 350 mg/Kg and 500 mg/Kg respectively on day one. Animals in group A and B acted as normal control and diabetic control respectively. All the animals except those in group A received simultaneous injection of streptozotocin 32 mg/Kg body weight intraperitoneally in a single dose. Fasting blood glucose was assessed and the animals weighed before starting the treatment, after 48 hours and at the end of the experimental period. Histological studies were carried out at the end of the study period. RESULTS: The blood glucose level and the final body weight of the animals in group C matched the values in diabetic control. Histologically the pancreas had generally reduced beta-cells mass (P < 0.001) with altered morphology. The animals in group D showed impaired glucose tolerance at 48 hours but were normoglycaemic at the end of the study period. There was some loss of beta-cells but a significant number of these cells (P < 0.05) showing normal morphology were saved. The animals in group E had normal number of beta-cells having normal morphological features. The final body weight and fasting blood glucose of these animals matched the values in normal control (group A). CONCLUSIONS: These data suggest that the optimum dose of nicotinamide in regard to prevention against the beta-cytotoxic effect of streptozotocin in albino rat is 500 mg/Kg body weight. PMID- 11873396 TI - Prevalence of hepatitis E virus (HEV) antibody in pregnant women of Karachi. AB - BACKGROUND: This study was aimed at determining the prevalence of HEV in Karachi. METHODS: Prevalence of hepatitis E antibody in pregnant women was carried out in 65 pregnant women. They had history of acute viral hepatitis while 15 control pregnant women were included having no history of jaundice. RESULTS: The statistical outcome of the present study indicates prevalence of HEV as 57% in pregnant women with jaundice. Majority of the patients (85%) were from lower socio-economic strata., with mean age of 25 years. All these patients had hemoglobin less than 10 gm%. The mean ALT level in HEV positive cases was 452 IU/L over a range of 102-5328. It was also observed that HEV affected more women in the last trimester (62%) and in primigravida (67%). It was observed that it was more common in last trimester and in primigravida. Majority of the patients (85%) were from lower socio-economic strata. CONCLUSIONS: The findings of the present study proved the presence of HEV in pregnant women and confirmed that HEV is endemic in Karachi. The study confirms that it occurs in last trimester and in young adult. PMID- 11873397 TI - Insulin therapy in NWFP Pakistan. PMID- 11873398 TI - Post-tonsillectomy and adenotonsillectomy morbidity & complications at District Headquarter Hospital Daggar. AB - BACKGROUND: Tonsillectomy or adenotonsillectomy is one of the commonly performed operations in ENT practice. Morbidity & complications associated with tonsillectomy and adenotonsillectomy include severe otalgia, pyrexia, odynophagia and haemorrhage. METHODS: A prospective study was undertaken from April 1996 to April 2001 to report the experience regarding postoperative morbidity/complications in tonsillectomy and/or adenotonsillectomy. A total of 1500 patients undergoing tonsillectomy or adenotonsillectom for chronic/recurrent tonsillitis or adenotonsillar hypertrophy were included. These patients received antibiotic in the form of amoxycillin and clavulanic acid and non-salicylate analgesics for 7 days postoperatively. 100 patients were lost to follow up. RESULTS: Seventy percent of patients were female and thirty percent males with maximum number of patients between the age of 11-22 years. Postoperative complications developed in 25 out of 1400 patients, 14 of them developed post operative bleeding and 9 developed postoperative infection. CONCLUSIONS: It was found that postoperative morbidity/complications were less in this study with regard to secondary haemorrhage, operative site infection, intensity & duration of postoperative pain, postoperative pyrexia and time for return to normal activities. PMID- 11873399 TI - Craniofacial alterations in adult rats after acute prenatal alcohol exposure. AB - BACKGROUND: Exposure during pregnancy to alcohol (ethanol) produces a number of adverse effects. One of them is fetal alcohol syndrome. The hallmark of fetal alcohol syndrome (FAS) is craniofacial dysmorphism and the changes in craniofacial measurement are dependent on the alcoholic dose and its time of exposure. Since prenatal ethanol exposure can alter craniofacial development in rodents and reliably produce long-term behavioral effect in them, the present study was designed to extend the same changes in the Sprague Dawley species. METHODS: The albino rat was studied to determine whether gestational exposure to alcohol (Ethanol) produces permanent craniofacial effect. On gestational day (GD7 10) 25% ethanol was injected intraperitoneally to pregnant rats. Various dimensions for skull and face of adult male rats were taken. RESULTS: Both vertical and coronal dimensions were altered in the exposed animals. CONCLUSIONS: This study demonstrates that exposure to ethanol on a critical gestational period produces permanent craniofacial defects. PMID- 11873400 TI - A rapid assessment study on prevalence of substance abuse disorders in metropolis Delhi. AB - BACKGROUND & OBJECTIVES: No studies in India have attempted to identify in the general population a 'dependent drug user' based on clinically used diagnostic criteria. This two point survey estimated the prevalence of substance use disorders and the change in the trends of drug use after a period of one year in metropolis Delhi. METHODS: A total of 72 colonies in five types of housing clusters were surveyed. The head of the household (HOH) was interviewed only for collecting information about family members (above 10 yr) on an instrument based on the DSM III R operationalised criteria. Trained non medical interviewers administered the precoded instrument to 6004 and 5599 HOH in the first and second surveys respectively. RESULTS: In the first survey, the prevalence of tobacco, alcohol, cannabis and opioids use among males was 27.6, 12.6, 0.3 and 0.4 per cent respectively. The rates were highest in resettlement clusters followed by urban villages, unauthorized, regularized and in 'others' clusters. The use rates remained unchanged during the re-survey. Dependent use (any drug) increased in the resettlement clusters only during the re-survey. INTERPRETATION & CONCLUSION: The results highlight that it is the legal drugs viz., tobacco and alcohol that could pose higher health and social consequences, both short and long term. The situation of illicit drug use (heroin) was higher in the resettlement clusters and urban villages. Need based programmes have to be evolved and executed to keep the drug dependent population stable. The rapid survey technique can be useful in developing countries like India, where resource crunch for survey research is acute. This technique is less costly, quicker to perform and can supplant traditional self-report methodologies. PMID- 11873401 TI - Modification of the multiplex PCR for unambiguous differentiation of the El Tor & classical biotypes of Vibrio cholerae O1. AB - BACKGROUND & OBJECTIVES: Biotyping of Vibrio cholerae O1 using multiplex PCR (ctxA-tcpA) exploits the nucleotide sequence differences of the major subunit protein of the toxin co-regulated pilus (TCP) gene (tcpA) to differentiate between the classical and El Tor biotypes. However, the presence of classical biotype specific tcpA amplicon with the El Tor strains often complicates the interpretation. The effect of PCR variables on the amplification of biotype specific tcpA in the multiplex PCR has been investigated. METHODS: Reference strains of toxigenic V. cholerae O1 belonging to classical and El Tor biotypes were selected to optimize the PCR variables for the unambiguous biotype determination by multiplex PCR. RESULTS: In the multiplex PCR assay, a reduction in the reaction volume from 100 microliters to 25 microliters and the annealing temperature of 64 degrees C, the El Tor strain produced ctxA amplicon (302 bp) along with tcpA amplicons of 618 bp and 472 bp which are specific for classical and El Tor tcpA respectively. The simplex PCR with biotype specific tcpA primer pairs showed the amplification of either 472 bp or 618 bp tcpA amplicon with El Tor template. With the classical biotype strain, the specific primer pair yielded tcpA amplicon of the expected size. Lowering of PCR annealing temperature from 64 to 60 degrees C resulted in the elimination of the amplification of the nonspecific tcpA amplicon with El Tor strain. INTERPRETATION & CONCLUSION: A comparison of the theoretical melting temperature (Tm) values of the reacting primers, and their alignment to the biotype specific tcpA revealed the basis of unambiguous biotyping of V. cholerae O1 at a PCR annealing temperature of 60 degrees C. PMID- 11873402 TI - Molecular comparison of toxigenic clinical & non-toxigenic environmental strains of Vibrio cholerae O1 Ogawa isolated during an outbreak of cholera in south India. AB - BACKGROUND & OBJECTIVES: While investigating a cholera outbreak in south India, toxigenic and nontoxigenic strains of Vibrio cholerae O1 were isolated from patients and from the environment, respectively. This study was performed to compare the genetic relatedness of the patient and environmental strains to determine clonal relationships among these strains and thereby determine the source of the cholera outbreak. METHODS: The 16 strains of V. cholerae isolated from hospitalized patients and 8 environmental V. cholerae strains isolated from the environment were phenotypically and genotypically characterized using a variety of standard techniques. RESULTS: Sixteen toxigenic clinical strains and 2 nontoxigenic environmental strains belonged to O1 serogroup, Ogawa serotype and El Tor biotype. The remaining 6 nontoxigenic environmental strains were classified as non-O1, non-O139 V. cholerae. The drug resistance pattern of the clinical and environmental strains of V. cholerae showed marked differences with the patient strains being resistant to more number of drugs as compared to the environmental strains. DNA fingerprinting of the strains showed considerable diversity between toxigenic clinical and nontoxigenic environmental O1 Ogawa isolates and between the O1 and non-O1, non-O139 isolates. INTERPRETATION & CONCLUSION: In this outbreak of cholera, the O1 strains of V. cholerae from clinical and environmental sources belonged to two different clones and the environmental strains could perhaps be the future cholera outbreak causing clones. PMID- 11873403 TI - A study on Staphylococcus aureus strains submitted to a reference laboratory. AB - BACKGROUND & OBJECTIVES: Staphylococcus aureus is regarded as one of the most devastating human pathogens. Recently there have been reports of increasing incidence of S. aureus strains resistant to methicillin (MRSA). A surveillance study was undertaken to record the occurrence of MRSA and to study the prevalence of various phage groups in India. METHODS: A total of 7574 strains of S. aureus received during 1992-98 at the National Staphylococcal Phage Typing Centre, New Delhi were tested for methicillin resistance and susceptibility to phages of the International basic set. The occurrence of various phage groups between MRSA and MSSA (methicillin sensitive S. aureus) was compared. Results were analyzed according to the geographical origin and source of isolation of the strains. RESULTS: The dominant phage group from different parts of the country was phage group III. Prevalence of phage group III among the MRSA and MSSA isolates was 62.32 and 33.95 per cent respectively. The highest isolation of phage group III strains was from nasal carriers (45.94%), phage group II strains from skin (8.74%), phage group I strains from blood (19.44%) and nontypable strains from the environment (80.68%). An increase in the occurrence of MRSA has been noticed from 9.83 per cent in 1992 to 45.44 per cent in 1998. INTERPRETATION & CONCLUSION: S. aureus strains of phage group III are prevalent in India. The increase in occurrence of MRSA indicates an alarming spread of these organisms. A constant monitoring is important to take appropriate and timely measures to control their spread. PMID- 11873404 TI - A study on some phenotypic virulence markers of enteropathogenic Escherichia coli. AB - BACKGROUND & OBJECTIVES: The problem of enteropathogenic Escherichia coli (EPEC) causing diarrhoea in infants exists in India. But often the enteropathogenic status is not based on adequate characterization. Hence there is a need for evaluating the serotyping being used to identify EPEC for its validity in the light of recent knowledge on phenotypic markers of virulence. This study was done to evaluate the EPEC isolates for two potential virulence factors namely entero adhesiveness with subsequent actin accumulation and verotoxin production. METHODS: Fifty consecutive EPEC strains identified by serotyping from stool samples of children with diarrhoea during January 1997 to June 1999 were studied for HEp-2 cell adherence, the fluorescent actin staining (FAS) characteristics of Hep-2 cells and vero cytotoxin production. RESULTS: Serotypes O55, O125 and O126 accounted for most of the isolates. In the Hep-2 assay, 72 per cent of the strains showed localised pattern of adherence and 22 per cent showed a mixed pattern of localised and diffuse adherence. In the FAS test 96 per cent strains showed typical staining while none of the strains produced verotoxin. INTERPRETATION & CONCLUSION: 'O' serogrouping appears to be still the simplest and an useful test for presumptive identification of EPEC. The FAS test for confirmation of EPEC was found to be very consistent in indicating EPEC. PMID- 11873405 TI - Evaluation of microscopic agglutination test as a diagnostic tool during acute stage of leptospirosis in high & low endemic areas. AB - BACKGROUND & OBJECTIVES: Making a diagnosis on the results of a single microscopic agglutination test (MAT) is difficult because of the uncertainties about the cut-off titre. The present study was conducted to determine the significant titre for a single MAT in areas of high and low endemicity for leptospirosis. METHODS: A total of 1944 serum samples were collected from healthy individuals and confirmed patients residing in areas of high and low endemicity. All the sera were screened by MAT using 10 live leptospiral strains as antigens. From the distribution of titres among healthy individuals and in patients, the sensitivity and specificity at different cut-off titres were calculated. Likelihood ratio positive (LR+), likelihood ratio negative (LR-), and LR+/LR- were calculated. Receiver operating characteristics (ROC) curves were plotted for the early and late stages of the disease in both the areas. RESULTS: The ROC plot was totally below the no benefit line during the first week of illness in high endemic area. During the second to fourth weeks it showed better characteristics and the best cut-off titre was 1:200, where the sensitivity was 93.4 per cent and specificity 74.7 per cent LR+ LR- ratio was 41.82 indicating reasonable separation between the positive and negative test results. In the other states the ROC plot was above the no benefit line even during the first week, the best cut-off being 1:50 where the sensitivity was 56.7 per cent and specificity was 90.6 per cent. During the second to fourth weeks the test showed the best characteristics in the low endemicity regions with an ROC curve having the ideal shape. Best cut-off was at 1:100 where the sensitivity was 96.6 per cent and specificity 94.8 per cent LR+ LR- ratio was 523.25 indicating a wide separation between the positive and negative test results. INTERPRETATION & CONCLUSION: MAT does not have any diagnostic value during the first week, particularly in high endemic areas. The best cut-off to be used will be 1:50 in low endemicity areas during the first week, 1:100 during the second to fourth week and 1:200 in high endemicity regions during the second to fourth weeks. PMID- 11873406 TI - [The state of anterior eye segment after posterior chamber intraocular lens implantation with transscleral fixation]. AB - PURPOSE: The aim of the study was to evaluate the state of anterior eye segment in postoperative period after PC-IOL transscleral fixation. MATERIAL AND METHODS: The material comprised 31 eyes in 31 patients, (11 women and 20 men), aged 20-80 years (average 61 years), who underwent the primary or secondary PC-IOL implantation with transscleral fixation. The examination was performed 10 to 14 months (average 12 months) after surgery. The clinical state of the corneas was evaluated using endothelial and confocal microscopy. The position of the IOL, its location and symmetry were evaluated with ultrasound biomicroscopy. RESULTS: The visual acuity ranged from 0.1 to 1.0. In 20 eyes (64%) the position of PC-IOL was correct. In the rest of cases the inappropriate position of implants was observed. The decrease of endothelial cells density was 13% in eyes with primary implantation and 7% in cases with secondary implantation of IOL. In the corneal endothelium, there were features of pleomorfism and polymegatism; in a few cases some deposits were observed. CONCLUSION: The implantation of posterior chamber intraocular lenses with transscleral fixation is recommended for some cases of primary or secondary problems with capsule support. The PC-IOL implantation with transscleral fixation can be the alternative method for the anterior IOL implantation. PMID- 11873407 TI - [Analyses of results of deep sclerectomy ab externo in open angle glaucoma based on own material]. AB - PURPOSE: The evaluation of postoperative results of the non-penetrating deep sclerectomy in primary open angle glaucoma. MATERIAL AND METHODS: From September 1999 to October 2000 the procedure was performed in 10 patients (15 eyes). There were 7 men and 3 women aged from 45 to 87 years. Preoperative IOPs ranged from 16 to 38 mm Hg. The follow up time ranged from 3 to 12 months. RESULTS: Success was considered an IOP of less than 20 mm Hg without medication in 87%. There were a few early postoperative complications included hypotony (9 patients), hyphema (1 patient), chorioidal detachment (3 patients). Late complication was bleb fibrosis in only 1 patient. Visual acuity and visual field were not worse than before surgery. CONCLUSIONS: Our experience confirms that the non penetrating deep sclerectomy appears to be a safe and effective procedure in open angle glaucoma. PMID- 11873408 TI - [Iron deposits in cornea in confocal microscope]. AB - PURPOSE: The study aimed to evaluate the iron deposits in corneas in confocal microscope. MATERIAL AND METHODS: The material comprised 16 eyes which underwent photorefractive keratectomy (PRK) procedure. The structure of corneas was evaluated between 3-10 years after PRK. The visual acuity after PRK was the same as the best corrected visual acuity before the procedure. The structure of corneas was evaluated in vivo using scanning slit confocal microscopy. The confocal images of corneas in patients after PRK were compared with confocal corneal images of patients with corneal scars (2 eyes), keratoconus (2 eyes), after radial keratotomy (RK) (2 eyes) and healthy patients. RESULTS: Within the central part of corneal epithelium and anterior part of stroma, the clusters of iron deposits were observed. They were round and produced different shapes. In the paracentral and peripheral part of corneas the subepithelial nerve plexus was detected. Beneath, the pattern of keratocytic nuclei, characteristic for state after PRK, was detected. In patients with corneal scars, keratoconus and after RK, the same clusters of deposits were detected. In cases of corneal scars, additionally high reflectivity of corneal structure was observed. CONCLUSIONS: The iron deposits in corneal structure arise in epithelium and anterior part of corneal stroma. The iron deposits which produce different shapes have no influence on visual acuity. PMID- 11873409 TI - [The causes of childhood blindness and visual impairment in Poland]. AB - AIM: To investigate the prevalence and the causes of childhood blindness and visual impairment in Poland. MATERIAL AND METHODS: The records of 3000 visually impaired children from the archives of Polish Association of Blinds and from the centers and schools for visually handicapped children from the years 1979-1999 have been reviewed. RESULTS: The number of the visually disabled children in Poland has increased by 70% in the recent 10 years. The main causes of blindness and serious visual loss in the years 1979-1999 are optic nerve atrophy (21.66%), retinopathy of prematurity (19.01%), cataracts (14.13%), high myopia (11.84%), congenital abnormalities (8.65%), retinal dystrophies (8.08%) and glaucoma (6.42%). Optic nerve atrophy occurred mainly in premature infants. There have been great changes in the epidemiology of blindness in the recent 20 years; the percentage of visually disabled children caused by ROP has increased from 8.1% to 54.5% and caused by optic nerve atrophy from 15.5% to 27.27%. The prevalence of other causes has decreased in the same time. CONCLUSION: The main activities required to control blindness in Poland are promotion of pregnancy and prematurity care and improvements in the early diagnosis and treatment of retinopathy of prematurity. PMID- 11873410 TI - [The results of clear lens extraction for anisometropia treatment in patients with high myopia and unilateral cataract]. AB - PURPOSE: To evaluate results of clear lens extraction for treatment of anisometropia in patients with bilateral high myopia and unilateral cataract. MATERIAL AND METHOD: 23 patients with bilateral high myopia who underwent clear lens extraction with PC IOL implantation to treat high anisometropia after cataract surgery in the other eye were observed in our study. Preoperative refractive error was from -8.0 D to -19.0 D (spherical equivalent). RESULTS: Anisometropia after operations in both eyes was from 0.0 D to 3.5 D. No serious complications after operations such as retinal detachment were observed. CONCLUSION: Clear lens extraction with PC IOL implantation in highly myopic eyes can be used for correction of high anisometropia after cataract surgery in the other eye. PMID- 11873411 TI - [Evaluation of tear film stability by means of laser interferometry]. AB - PURPOSE: The paper presents a new method for evaluation of the tear film stability on the human eye. METHODS: The tear film distribution on the cornea is measured by the lateral shearing interference technique. The eye is kept open during approximately a two-minute recording, when the blinking has to be prevented. Continuous recording and viewing of interferograms enables registration of the changes in disturbances of interference fringes during elapsed time. The changes in fringes are caused by evaporation of tears from the ocular surface and appearance of the breakups. CONCLUSIONS: The noninvasive tear breakup time (NITBUT) can be evaluated by comparing the recorded consecutive interferograms. PMID- 11873412 TI - [Evaluation of the surface of the new intraocular lenses in the scanning electron microscope]. AB - PURPOSE: To evaluate the surface of the new PC IOLs commercially available in Poland in 2000. MATERIAL AND METHODS: Representative samples of new posterior chamber IOLs produced by 6 different companies (Alcon, Lensita, Medicontur, Opsia, Rayner, Storz), 5 of each, underwent surface examination with Novoscan 30 scanning electron microscope. RESULTS: Although, in general, smooth surface of optic and haptic parts were observed, three samples with irregularities were found. CONCLUSIONS: Comparing to previous evaluation performed in 1994, significant improvement in quality of IOLs surface was noted. No considerable differences in this field between above mentioned producers were observed. PMID- 11873413 TI - [The use of indocyanine green angiography in differential diagnosis in intraocular tumors]. AB - The author presents current opinions about the usefulness of indocyanine green angiography in differential diagnosis of choroidal tumors: melanomas, naevi, haemangiomas and choroidal metastasis. PMID- 11873414 TI - [Usefulness of visual evoked potentials in visual acuity examination]. AB - Determination of visual acuity by means of VEP is carried out usually by the calculation of the amplitude function response from the value of pattern single element. That purpose is achieved by carrying out several tests of transient VEP performed each time with stimulation, using the pattern of different size of individual elements or changing sweep the following sizes of the pattern single element. Concentration of a patient is necessary during any examination and therefore in clinical practise the second of the listed methods is preferred because of its short duration. Correlation factor of visual acuity, determined by means of VEP and Snellen tests, differs depending on the examined group from about 0.4 to 0.9 and it is the lowest in case of ophthalmological disorders connected with optic nerve. Fixing of VEP optimal parameters when evaluating the visual acuity remains an open matter. PMID- 11873415 TI - [The role od adhesion molecules in the pathogenesis of Graves ophthalmopathy]. AB - Graves' ophthalmopathy (GO) is an autoimmune condition characterized by mononuclear cell infiltration of the extraocular muscles and/or orbital connective tissue. Adhesion molecules play an important role in the initiation and maintenance of the inflammatory immune process. Cellular activation and local expression of adhesion molecules lead to leucocyte recruitment, migration to inflammatory sites and targeting in the extravascular space. Vascular endothelium in retroocular connective tissues of patients with GO is strongly positive for EMAL-1 and VCAM-1, whereas VCAM-1 immunoreactivity is minimal and ELAM-1 immunoreactivity is generally absent in normal retroocular tissue. Interactions between matched activated T lymphocytes and orbital endothelial cells are mediated by integrin dependent ICAM-1/LFA-1 and VCAM-1/VLA-4 pathways and reveal marked differences when comparing GO orbital endothelial cells to normal ones. Higher soluble ICAM-1 volumes in patients with Graves' disease with GO than those in patients with Graves' disease without ophthalmopathy can reflect the degree of inflammatory activity. Increased soluble ELAM-1 concentration only in patients with GO suggests that soluble ELAM-1 could be a specific marker of endothelium activation in GO. PMID- 11873416 TI - [The effect of serotonin on flash visual evoked potential in the rat prenatally exposed to cadmium]. AB - AIM: The purpose of this paper was to find out any influence of cadmium (Cd) on the effect of serotonin (5-HT) in the central nervous system (CNS). METHOD: 18 Wistar albino strain female rats were divided into 3 groups: 6 received 5 ppm, 6 one 50 ppm cadmium in tap water since time of conception and during next 21 days after delivery. Control group of 6 rats received tap water only. Newborns were examined when they were 3 to 6 months old. Flash visual evoked potential (FVEP) was recorded before and after injection of 10 microliters saline and then 5-HT into the right lateral brain ventricle. Two doses of 5-HT 125 and 250 nmols were used. Amplitudes of the first deep negative wave (N1) and the next positive one (P1) were measured from isoelectric line to peaks. For statistic analysis the t test of Student was used with statistical significance by p < 0.05. RESULTS: Shortened latencies (89-99%) of the peaks N1 and P1 of FVEP in the control and Cd 50 ppm groups after both doses of 5-HT were observed, however, they slightly prolonged (103-105%) in the Cd 5 ppm group. The highly significant increase of amplitude of the waves N1 and P1 in all observed groups was received. The only differences were observed in the Cd 5 ppm group; the amplitude of P1 peak was of the same value (100-101%) after 125 nmols and decreased (91%) after 250 nmols of 5-HT compared to the initial values in this group. CONCLUSION: Cadmium increases the serotonin sensibility in the CNS. PMID- 11873417 TI - [Histopathological evaluation of orbital tissue encircling intraorbital implant made of glassy carbon and its influence on basic blood parameters in a rabbit]. AB - PURPOSE: The purpose of the study is to present histopathological evaluation of orbital tissue encircling intraorbital implant and to present influence of glassy carbon on blood parameters of rabbits. MATERIAL AND METHODS: We examined 20 rabbits (Oryctolagus Cuniculus) and 2 pigs (Suis Domesticus) with carbon orbital implants. Periodically the experimental animals were killed 5, 20, 35, 60, 100, 150 days after implantation of glassy carbon into the orbit. Encircling tissue was histopathologically examined. The blood was taken to analysis on 0, 5, 20, 35, 60, 100, 150 day after procedure. RESULTS: Histopathological specimens of rabbits did not show intolerance reaction of glassy carbon. Histopathological picture was typical of the "foreign body" reaction. Histopathological pictures of pigs did not show any reaction to implant. There was no deviation in blood parameters. CONCLUSIONS: We suggest that intraorbital glassy carbon implants do not induct intolerance reactions to orbital tissues. Glassy carbon implant does not have any influence on basic blood parameters of rabbits. PMID- 11873418 TI - [Use of amnion in the treatment of anterior segment diseases of the eye]. AB - PURPOSE: The aim of this study is to present the results of treating the anterior segment of the eye with amnion membrane transplantation (AMT). MATERIALS AND METHODS: 16 patients were treated with amnion membrane transplantation because of: pemphigoid ocularis, combustio chaemica, ulcus corneae, keratopathia bullosa, descemetocele, symblepharon, graft disease. Histopathological investigations were performed by the use of impression cytology. RESULTS: In 4 patients we obtained only temporary improvement. In 12 patients the treatment was successful with complete reepithelialization of the cornea and the conjunctiva. The impression cytology study showed an intensive regeneration of the corneal and conjunctival epithelium. PMID- 11873420 TI - Preventing AIDS: men matter most. PMID- 11873419 TI - [Treatment of corneal ulcerations with use of the amniotic membrane]. AB - Corneal ulcerations may cause complications such as, for example, the loss of transparency, descemetocele or perforation of the cornea. Widely used therapies do not always bring expected results. Recently the amniotic membrane has been applied for the treatment of corneal ulceration. PURPOSE: The purpose of the present study was to evaluate the efficacy of the amniotic membrane suturing over the locations of corneal ulcers. MATERIAL AND METHODS: The amniotic membrane was sutured over the locations of corneal ulcers in 9 eyes of 9 patients suffering from ulcers which did not regress with traditional therapeutic methods. After meticulous debridement of the ulcer floor and border area the amniotic membrane was sutured to the cornea with a single 10.0 Nylon suture (around the ulcer area), and additionally suture were fixed to the sclera. Following the procedure contact lenses were placed over the area for protection. The observation period was 6 months. Follow-up examinations were performed regularly during the first seven postoperative days, then after 2, 4 and 12 weeks; the final follow-up took place 6 months after the procedure. RESULTS: In all patients we observed healing of the corneal defect, improvement of visual acuity and regression of the active inflammatory process. After this procedure the subjective symptoms subsided. CONCLUSION: The amniotic membrane can be used for the treatment of corneal ulceration of various etiology. PMID- 11873421 TI - Concentration of heavy metals in drinking water of different localities in district east Karachi. AB - BACKGROUND: Several heavy metals are present in drinking water that play important roles in the body provided their level remains within the specified range recommended by WHO. But now due to the industrialization and rapid urbanization, the problems of pollution have surfaced. This study was designed to ascertain the contents of some heavy metals and then their variations if any in drinking water in different localities of district East of Karachi, Pakistan. METHODS: Drinking water samples were collected from different sources and localities of district East of Karachi. The concentration of the heavy metals i.e. Lead, Arsenic, Copper, Iron, Mercury, Chromium, Manganese, Nickel, Cadmium and Zinc were determined by Atomic Absorption Spectrophotometry. PH was estimated by pH meter. Total dissolved solids (TDS) were calculated by formula. These concentrations of heavy metals, pH and TDS were compared with the standards set by WHO. RESULTS: Concentrations of lead and nickel were found to be significantly elevated as compared to WHO recommended levels in all the three sources of water (Piped water, Hand pump water and Tanker water supply). Chromium was found to be raised in hand pump water. Arsenic and Mercury were not detected in any source of water. Copper, iron, manganese, cadmium and zinc were found to be within the safe limits in all the three sources of water. pH was found to be within the range of WHO recommended level in all the three sources of water. TDS was found to be elevated in hand pump water and tanker water. CONCLUSION: Concentrations of lead and nickel were found to be significantly elevated as compared to WHO recommended levels in all the three sources of water in district East of Karachi. PMID- 11873422 TI - Prevalence and pattern of congenital heart disease in Hazara. AB - BACKGROUND: The incidence and particularly the pattern of congenital heart disease may vary in different geographical locations, but the extent to which such reported variations are attributable to differences in genetic predisposition, environmental factors, or in study methodology and diagnostic precision remains uncertain. There is, therefore, a continuing need for studies on various aspects of congenital heart disease, in different communities and races. METHODS: This study was done on 188 consecutive cardiac patients reporting at Ayub Teaching Hospital, Abbottabad, Pakistan from June 1998 to June 2000. RESULTS: 114 of 188 cardiac patients studied had congenital heart disease. Both sexes were equally affected. Ventricular septal defects were the most common lesions (relative frequency 61.4%), followed by Tetrology of Fallots, ASD and PDA with a relative frequency of 8.77%. The detection rate under one month was 28.07% and at one year 75.43%. CONCLUSION: There is therefore, a need for increased awareness, especially among primary health and other front line doctors for earlier case detection. PMID- 11873423 TI - Stat tests service and quality control at Hamad Medical Corporation Doha. AB - BACKGROUND: The recognition and validity of laboratory results can better be judged by its internal and external quality control systems. Here we report an internal quality check of the stat service provided to the inpatients at Hamad Medical Corporation, Doha, State of Qatar. METHODS: Periodic internal quality control of nine chemistry analytes, BUN, Na, K, Cl, Ca, Co2, Glucose, Creatinine and Bilirubin were made on auto-analyzer Astra VIII. BIORAD Normal and Abnormal (Chemistry Control) and immunoassay level 1, 2 and 3 were employed. All results falling with in 2 SD were accepted. RESULTS: 2 SD% for chemistry Normal and Abnormal ranged from 61.5% to 85.5% and 92.0% to 98.8% and for hormones Immunoassay level 1, 2, and 3 from 92.7% to 100% respectively. PMID- 11873424 TI - Comparison of aztreonam against other antibiotics used in urinary tract infections. AB - BACKGROUND: Urinary tract infection is a very common problem in adults as well as in children. There is always need for the right antibiotic to be chosen for treatment. METHODS: This study was conducted at Microbiology section in Pathology department of Ayub Medical College, Abbottabad, Pakistan. In this study the aztronam which is only effective against gram negative bacilli has been compared with other conventionally used antibiotics, ampicillin, cotrimoxazole, minocyclin, pipemedic acid, nalidixic acid, norfloxacin, ciprofloxacin, gentamicin and ceftriaxon. RESULTS: Total gram negative isolates were 342. Out of this 76.6% (262) were E. coli, klebsiella pneumoniae 14.3% (49), proteus species 5.2%(18) and pseudomonas aeruginosa were 3.8% (13). The aztreonam showed 78% sensitivity against gram negative bacilli which is better than norfloxacin which showed 62.2% sensitivity. PMID- 11873425 TI - A study of relationship of ABO blood groups with myocardial infarction and angina pectoris. AB - BACKGROUND: This is a comprehensive report that has determined the occurrence of Myocardial infarction and Angina pectoris in "ABO" blood group system among patients with coronary artery disease in some areas of Sindh province of Pakistan. METHODS: Three hundred patients with Coronary Disease (CAD) were selected from cardiology wards of LMC hospital Hyderabad, DMC hospital Karachi and PMC hospital Nawabshah. The patients were separated into two categories: myocardial infarction and angina pectoris. The patients with old myocardial infarction were also included. A careful history was taken, suggesting myocardial infarction (MI) or angina from a standard WHO (Rose) chest pain, and a electrocardiogram showing evidence of possible myocardial infarction and angina, and the patient's recall of a doctor's diagnosis of M.I or angina. ABO blood grouping of above patients was done by simple agglutination method. RESULTS: The blood group "A" was the commonest among myocardial infarction and angina pectoris patients while these diseases were least in blood group "O" patients. CONCLUSIONS: This comparison shows the existence of a direct relation between blood group antigens and coronary artery disease. It is therefore of great importance for future genetic studies, as present report and our previous studies give clear picture of excess and deficit of CAD in particular blood groups of "ABO" system. This may be due to some special genetic makeup. PMID- 11873426 TI - Effect of oral and injectable contraceptives on serum electrolytes, weight and blood pressure. AB - BACKGROUND: Since the introduction of oral and injectable contraceptives, many metabolic side effects have been reported. The use of oral contraceptive has been shown to be associated with increased level of serum electrolytes, but decrease in levels of serum electrolytes has been reported in women using injectable contraceptives. This study evaluated serum electrolytes, weight and blood pressure in women taking oral and injectable contraceptives because these changes have adverse effects on the health and economy of the subject. METHODS: Serum electrolytes, weight and blood pressure were measured in 50 women taking oral contraceptive (Lofeminal) and 50 women taking injectable contraceptives (Depomedroxy progesterone acetate and Norigest). The same women served as control before starting these contraceptives. RESULTS: There was a significant increase in serum sodium and chloride but significant decrease in serum potassium and bicarbonate level in women taking oral contraceptive. There was non-significant decrease in serum electrolytes in women taking injectable contraceptives. There was significant increase in weight and blood pressure of women taking these contraceptives. PMID- 11873427 TI - Cornucopia of cultured Schwann cells: where is the best source in albino rats? AB - BACKGROUND: Schwann Cells (SC) are multifunctional glia cells with diversified biological potentials. To study their biological activities, ample amount of cultured SC is a pre-requisite. Many sources have been exploited in the past for SC culture but unfortunately no attempt has been made to find the richest source. METHODS: To make an attempt in this direction, we cultured SC under identical conditions from various neural sites in the neonate & young SD albino rats. Fresh, cryopreserved & pre-stimulated neural tissues have been compared for total & % of SC/unit tissue from the combined data of living & immunostained cultures. RESULTS: Our data shows that the upper limb nerves have not only quantitative advantage (2.35 x 10(4) cells/mg tissue) but also have significant qualitative edge (P < .05-.01) over the remaining sources. It can be concluded from our data that upper limb nerves are the best available source for SC/unit tissue. PMID- 11873428 TI - The incidence of non ulcer dyspepsia and its response to treatment. AB - BACKGROUND: This study was undertaken to identify Non Ulcer Dyspepsia (NUD) in young patients presenting with peptic ulcer like symptoms. Their response to treatment was also determined. METHODS: 100 patients between the ages of 20-40 of both sexes were included in the study. Investigations done were Chest X-ray Plain Abdominal X-ray, ECG, ESR, LFTs Amylase Abdominal Ultrasound and upper GI endoscopy. All patients then received 6-week therapy with Omeprazole and Helicobacter Pylori eradication with Clarithromycin and Tinidazole. RESULTS: NUD was by far the most common cause (59%) in both the sexes. Response to therapy was unsatisfactory. CONCLUSIONS: This relatively newly discovered clinical entity should be kept in mind while dealing with patients presenting with refractory dyspeptic symptoms. PMID- 11873430 TI - Nephrotic syndrome: minimal change disease. PMID- 11873429 TI - Dermatofibrosarcoma protuberans--a rare variant of soft tissue sarcoma of skin. An experience of two cases at Ayub Teaching Hospital Complex (ATHC), Abbottabad. PMID- 11873431 TI - Prevalence of mental health problems in acne patients. AB - BACKGROUND: Acne is the most common of all skin diseases. The condition affects young people at the time when they are undergoing maximum social and physical change and can produce significant psychological scarring. This study was done to determine the Mental Health problem in Patients suffering from acne and to determine significant difference of mental health problems in acne patients in comparison to seborrhic dermatitis patients as a control group. METHODS: 50 subjects with acne and 50 with seborrhic skin problems (13-25 yrs age) were included in this study through Psychiatry OPD of Lady Reading Hospital, Peshawar. The period of study was from March 1998 to August 1998. The patients were assessed for psychiatric problems by using WHO Guide to mental health in primary care ICD 10 chapter V primary care. The statistical significance was determined by chi square. RESULTS: The data shows that out of 50 acne patients 19 (38%) were suffering from Depression, with a female predominance. The control group (seborrhic) presented lower prevalence of Depression i.e. 57% in females. Similarly Anxiety problem in particular social anxiety were overall more in acne group (34%) as compared to seborrhea group (10%). A statistically significant difference was observed between the two groups indicating that depression and social anxiety was more in the acne group. A consistent finding in the study over six months depicted the difference between gender i.e. more females presenting with depression. Total Depressed Patients were 19, while total with suicidal Thoughts were 4 (21%). CONCLUSION: The importance of this study is to highlight depression among such patients. PMID- 11873432 TI - Impact of workshops on Themes of World AIDS Days. AB - BACKGROUND: Abbottonians Medical Association, Abbottabad, Pakistan in collaboration with UNAIDS Pakistan arranged two Workshops, the first in 1999, the Theme of which was "Listen, Learn, Live", and the second in the year 2000 with "Men Make Difference" as the Theme. The aim of these workshops was to create awareness amongst the participants regarding HIV/AIDS. OBJECTIVES: We used the data of the above-mentioned workshops to analyze the gain in knowledge and attitudes of the participants, with an idea to develop recommendations for the use of such workshops for increasing the awareness regarding HIV/AIDS. METHODS: The data of the two above-mentioned workshops were used to assess the percentage gain in knowledge and attitudes of the participants. We also compared our results with the contemporary studies and developed our recommendations. RESULTS: The results show highly significant (p < .001) increase in the HIV/AIDS related knowledge and attitudes in the participants of these workshops. This difference was more significant in the teachers as compared to the students, showing a higher level of awareness in the youngsters. The net gain in the knowledge was very significant. The knowledge about HIV spread increased by 28.96% for the students and 19.54% for the teachers, while the net gain in attitudes was 36.16% for the students and 71.43% in the teachers. CONCLUSIONS: Short workshops on the Themes of World AIDS Days are very beneficial for improvement in the level of awareness for HIV/AIDS prevention and control. PMID- 11873434 TI - [Chronic pulmonary thromboembolism originating from left forearm deep vein thrombosis: a case report]. AB - Upper extremity deep venous thromboembolism has become increasingly common due to the use of subclavian venous access. However, forearm deep venous thrombosis is rare. We report a case of chronic pulmonary thromboembolism originating from left forearm deep venous thrombosis. A 66-year-old woman was admitted to our hospital because of dyspnea, which had worsened over the previous 2 months. Echocardiography showed severe right ventricular enlargement and pulmonary hypertension. Perfusion lung scintigraphy revealed multisegmental perfusion defects. No likely factors responsible for hypercoagulability were found. The patient had a history of left radial fracture and had been treated with external fixation. Venography of the upper and lower extremities revealed total occlusion of the deep veins of the left forearm with collateral vessels, but no evidence of other upper or lower extremity venous thrombosis. PMID- 11873433 TI - P wave signal averaged electrocardiography in patients undergoing the Fontan operation. AB - OBJECTIVES: Supraventricular arrhythmias are one of the most common and fatal sequelae of the Fontan operation. P wave triggered signal averaged electrocardiography was performed in patients undergoing the Fontan operation to evaluate the presence of atrial degeneration, and to clarify which factors affected the development of atrial arrhythmias. METHODS: P wave triggered signal averaged electrocardiography was recorded in 14 patients after the Fontan-type operation (conventional atriopulmonary connection in 5 and total cavopulmonary connection in 9) and 15 healthy controls. The duration and area of the filtered P wave, and the signal magnitudes (M20, M30) at 20 Hz and 30 Hz obtained from the frequency domain analysis of the P wave (M20, M30) were evaluated and compared with the hemodynamic data. RESULTS: The duration and area of the filtered P wave, M20 and M30 in patients after atriopulmonary connection were significantly greater than in those after total cavopulmonary connection and the control subjects (p < 0.05). M20 was significantly greater in patients after total cavopulmonary connection than in the control subjects. Right atrial volume in patients after atriopulmonary connection was significantly (p < 0.001) larger than in patients after total cavopulmonary connection (p < 0.05). There were no significant differences in other indices including atrial pressure between the two groups. CONCLUSIONS: Our results suggest that the substrate for atrial arrhythmias such as atrial myocardial degeneration and fibrosis is frequently present in patients after the Fontan operation, especially after atriopulmonary connection. Thus, the enlarged right atrium may be involved in the presence of a substrate for atrial arrhythmias. The developmental risk for late atrial arrhythmias seems to be present even in patients after total cavopulmonary connection. PMID- 11873435 TI - Challenging case of pulse infusion thrombolysis using a unique pump system for a patient with deep vein thrombosis: a case report. AB - A 69-year-old man presented with chronic deep vein thrombosis due to massive thrombi extending from the inferior vena cava to both femoral veins. He had undergone surgery for prostatic cancer in 1991, and since then he had been taking an artificial estrogen agent. He was successfully treated by pulse infusion thrombolysis using a unique pump system, which we have developed, without complication. PMID- 11873436 TI - [A 78-year-old man with left hydronephrosis. Atherosclerotic abdominal aorta and retroperitoneal fibrosis]. PMID- 11873437 TI - [A 27-year-old woman suffering repeated syncopal attacks. Congenital discrete subaortic stenosis Kelly, type I: membranous type]. PMID- 11873438 TI - [Left ventricular peak systolic pressure/end-systolic volume ratio change after dobutamine infusion for predicting left ventricular contractile reserve: comparison with Emax]. AB - OBJECTIVES: Genuine left ventricular contractile function is difficult to assess in the clinical setting. Left ventricular peak systolic pressure/end-systolic volume (Pps/Ves) ratio may be misleading because this index takes no account of the left ventricular end-systolic point and V0 intercept in the pressure-volume relation geographic curve. End-systolic pressure-volume relation and maximum chamber elastance derived from left ventricular pressure-volume loops can provide reliable estimates of contractile function. However, the feasibility of this technique for clinical purposes is limited, because it requires instantaneous measurement of left ventricular pressure and volume. This study assessed the feasibility of using Pps/Ves ratio for predicting the left ventricular contractile reserve by direct comparison with maximum elastance (Emax) derived from left ventricular pressure-volume loops. METHODS: Studies were undertaken in 18 consecutive patients aged 60 +/- 9 years who underwent cardiac catheterization. On-line instantaneous left ventricular volume was derived from the acoustic quantification method by transthoracic echocardiography. Pps was determined by pressure manometer tipped wire and Ves was measured automatically from acoustic quantification software in an ultrasound system. Pps/Ves was compared with Emax derived from each simultaneous pressure-volume loop during inferior vena caval occlusion before and after dobutamine infusion. Emax was determined as the slope of end-systolic points for each loop with the use of an automated iterative linear regression technique. Left ventricular contractile reserve was assessed by evaluating its functional response to 10 micrograms/kg/min of dobutamine infusion. RESULTS: Pps/Ves showed significant correlation with Emax in all patients (r = 0.70, p < 0.0001). However, scattered distribution of V0 value differences were noted. Contractile reserve (Pps/Ves) showed strong correlation with contractile reserve (Emax) despite V0 value differences (r = 0.927, p < 0.0001). CONCLUSIONS: Pps/Ves change after dobutamine infusion may minimize individual V0 distribution. This simple index could be used to evaluate left ventricular systolic performance without requiring the left ventricular pressure-volume relationship and volume unloading maneuver. PMID- 11873439 TI - [Tei index evaluated by M-mode echocardiography in patients with dilated cardiomyopathy]. AB - OBJECTIVES: The Tei index obtained by the Doppler method is effective for analysis of global cardiac dysfunction. M-mode recordings of the aortic valve and mitral valve can also provide this index. METHODS: Forty-two patients with cardiomyopathy (idiopathic 15 and ischemic 27) with left ventricular ejection fraction < 40%, and 85 normal subjects were enrolled in the study. The M-mode Tei index (Tei-M) was obtained from the ejection time and closing period of the mitral valve in the same manner as the Tei index obtained by the Doppler method (Tei-D). RESULTS: The values of Tei-D and Tei-M were closely correlated (r = 0.75, p < 0.0001). Using index > or = 0.45 as the cut off value, patients with cardiomyopathy were identified with a sensitivity of 86% and a specificity of 78% by Tei-D, and 67% and 73% by Tei-M, respectively. In patients with mitral B-bump on M-mode echogram (n = 10), there was no mitral inflow during B-bump. Thus, Tei M was smaller than Tei-D in these patients. Using the new criteria of Tei-M > or = 0.45 and presence of B-bump, patients with cardiomyopathy were identified with a sensitivity of 86% and specificity 73% by Tei-M which were comparable with Tei D. CONCLUSIONS: The Tei-M is a simple and effective index for the analysis of cardiac dysfunction in patients with cardiomyopathy. PMID- 11873440 TI - [Relationship between long-term preventive efficacy of cibenzoline and atrial natriuretic peptide in patients with paroxysmal atrial fibrillation]. AB - OBJECTIVES: The relationship between the long-term efficacy of the antiarrhythmic agent cibenzoline in preventing lone paroxysmal atrial fibrillation (PAf) and plasma concentrations of atrial natriuretic peptide (ANP) and catecholamine was investigated during sinus rhythm and PAf. METHODS: Plasma concentrations of ANP, epinephrine, norepinephrine and dopamine during sinus rhythm and PAf were measured in 87 patients (70 men, 17 women, mean age 64 +/- 11 years) with lone PAf. All patients received cibenzoline (300 mg/day) after cardioversion, and they were divided into the no recurrence group (n = 28) and the recurrence group (n = 59). Mean follow-up period was 41 +/- 29 months. RESULTS: The plasma concentration of ANP was significantly higher during PAf (110.2 +/- 65.0 pg/ml) than during sinus rhythm (39.9 +/- 27.8 pg/ml, p < 0.01) for all patients. The concentrations of epinephrine, norepinephrine and dopamine during PAf were all similar to those during sinus rhythm. Patient characteristics showed no statistically significant difference between the no recurrence and recurrence groups. In the recurrence group, the incidence of thromboembolism was significantly higher (30.5% vs 10.7%) and the period of PAf was significantly longer (26.8 +/- 43.6 vs 12.4 +/- 21.2 months) than in the no recurrence group (both, p < 0.05). The plasma concentrations of ANP during sinus rhythm were similar in the no recurrence group (33.1 +/- 20.1 pg/ml) and the recurrence group (43.5 +/- 30.3 pg/ml), but was significantly higher during PAf in the no recurrence group (142.6 +/- 76.5 pg/ml) than in the recurrence group (95.8 +/- 54.2 pg/ml, p < 0.01). The ratio of ANP level during PAf to that during sinus rhythm in the no recurrence group (5.0 +/- 2.5) was significantly greater than that in the recurrence group (3.2 +/- 2.5, p < 0.01). CONCLUSIONS: Patients without recurrence of PAf under treatment with cibenzoline have preserved capacity of ANP secretion compared with patients with recurrence. PMID- 11873442 TI - Health care privacy: exceptions to the rule preparing for the brave new world, Part III. AB - As physicians prepare for implementation of HIPAA, there will be times when they are asked to and may be required to release individual health information without patient consent and authorization. This third article in a four-part series highlights the situations in which a doctor may release health information without a patient's knowledge. PMID- 11873441 TI - The agenda shifts to health care protection. PMID- 11873443 TI - Creating a more efficient physician practice: six steps toward turbo-charged practice revenues. AB - Health care providers face reimbursement and expense constraints. They will be helped in streamlining their operations by adopting financial benchmarks and following a six-step formulation to improve and assess their efficiency and effectiveness. These include: revenue and cost analysis; improved staffing and training procedures, attention to scheduling; developing functional patient intake procedures; selecting appropriate information systems; and evaluating the patient encounter by surveys and the examination of financial results. PMID- 11873444 TI - Measuring patient loss. AB - Few medical practices collect patient loss data. Yet patient defection has an enormous impact on practice profitability. Not only is the information on patient defection painful to analyze, but also difficult to capture. Patient requests for transfer of medical records are an insufficient measure of patient loss. Billing services are the likeliest source of patient loss data, but they have varying capabilities to provide this information, the information is often flawed, and each time a practice switches to a new billing service, it loses the ability to produce patient retention and defection data for a period of time. The previous article in this series detailed the costs of defection of patients to medical practice profitability together with methods to help retain patients. However, most practices do not or cannot quantify patient loss. This article concentrates on the problems of tracking defections and offers a possible solution through a focus on the data provided by billing systems. PMID- 11873445 TI - Practice re-engineering through the use of benchmarks: Part 1. AB - Benchmarking can be a useful tool to assist practices in successfully re engineering their practices to achieve performance improvement. Unfortunately, although many practices identify benchmarks, they often are not integrated into day-to-day operations or used to create accountability or re-engineer processes. Part 1 of this series describes the nine-step benchmarking process and discusses how to successfully use the process to improve performance through process re engineering. Part 2 will identify key indicators to measure performance and explain how to integrate their use into the practice's daily operations. PMID- 11873446 TI - How to increase economic returns and reduce liability exposure: Part 2. PMID- 11873447 TI - Verifying a job applicant's factual credentials. AB - Nearly one-third of all job applicants may make unjustified claims, exaggerate, omit important information, or lie in their resumes, employment applications, or interviews. Checking credentials before you hire is the best way to protect your staff and patients and to save time, money, and effort. In this article, the author identifies the most common job application lies and pinpoints the three most common resume/application red flags. In addition, the article suggests specific information that the applicant should supply in the application to help the employer get to the truth. It offers practical advice for verifying academic credentials, employment history, and awards and honors. Finally, this article provides useful suggestions for avoiding discrimination suits in hiring practices and techniques for coping with employees who have lied to you. PMID- 11873448 TI - Phone technology systems help practices communicate with patients. AB - Modern computer technology provides practices with modalities to communicate directly with patients at low cost. This article describes several TeleVox systems that allow managers to confirm appointments, issue recalls, and transmit laboratory and other data. Other systems allow patients to leave messages for their providers more directly and also allow practices to supply educational information on medical topics while patients are on hold. PMID- 11873449 TI - Are you committing health care fraud under the False Claims Act? The answer may not be as simple as you think. AB - Health care fraud has a significant financial effect on the operations of the American health care system. Governments and insurers are intent on quashing such fraud and have instituted massive efforts to indict and punish offenders as well as recover major monetary awards. This article provides examples of fraudulent practices and details the definitions of fraud and abuse. In addition, it characterizes the concept of qui tam that allows whistleblowers to share in any financial awards. Finally, it highlights the importance of proper coding as a means of reducing the suspicion of fraud or abuse. PMID- 11873450 TI - Life insurance and its effect on estate taxation. AB - Estate tax laws are subject to constant changes and revisions. Fortunately for most physicians, this ultimate liability has been drastically reduced, but it also has become much more complex. The key is to plan proactively with an understanding of the most current estate tax savings strategies. PMID- 11873451 TI - First Health: a new model for health benefits management. PMID- 11873452 TI - Trends in the consumer price index. PMID- 11873453 TI - Using the Internet to deliver health care value. AB - Beyond the popular uses of the Internet by HCOs--for recruiting employees, purchasing supplies and promoting brand and service awareness--lie a host of opportunities to add value to consumers online. All HCO programs can be enhanced through online initiatives: wellness and health promotion, risk reduction, prevention, early detection, symptom management, life event management, acute treatment and rehabilitation, disease management and end-of-life improvement. And beyond online initiatives in each of these categories lies the potential to use the Net for reminding consumers, individually and collectively, of the health and quality of life benefits they are gaining, thereby adding to the HCO's marketing and PR success. PMID- 11873454 TI - Physicians' adoption of information technology: a consumer behavior approach. AB - Studies report physician resistance to information technology in a time when the practice of medicine could benefit from technological support. Anecdotally, it is suspected that lack of training, discomfort with technological innovations, a perceived shift in the doctor/patient relationship, or medical/legal issues may account for this circumstance. Empirical studies attribute this lag to age, personality factors, behavioral issues, and occupational influences. This paper integrates the information technology and consumer behavior literatures to discuss physicians' acceptance, adoption, and application of IT. PMID- 11873455 TI - Redesigning geriatric healthcare: how cross-functional teams and process improvement provide a competitive advantage. AB - This study examines the consequences of adding a geriatric subacute unit to the traditional health care mix offered by a nonprofit hospital. Historically, geriatric health care offerings have been limited to either acute care units or long-term care facilities. The study's findings demonstrate that the addition of a subacute unit that is operated by an interdisciplinary team is a competitively rational move for two reasons. First, it provides a continuum of care that integrates services and departments, thereby reducing costs. Second, it provides a supportive environment for patients and their families. As a consequence patients have a higher probability of returning home than patients who are assigned to more traditional modes of care. PMID- 11873456 TI - Taking your medicine: relational steps to improving patient compliance. AB - Patient non-compliance with physicians' instructions is a major problem that costs billions of dollars each year. This study supports a significant role for communication, both as a form of information exchange and social support, and participative decision-making in improving patient compliance. These results, based on structural equation modeling, also support the interaction of communication and participative decision-making positively affecting compliance. Results suggest that one-way communication from physician to patient and patient education will not solve compliance problems by themselves. Instead the solution revolves around open, bi-directional information exchange, active listening by both parties, and truly informed consent on the part of patients. PMID- 11873457 TI - Determinant attributes in nursing home choice: profiling attribute segments. AB - The majority of research of nursing home choice focuses on attribute importance. In comparison, this study identifies attributes that determine choice. Utilizing dual-questioning methodology, 177 nursing home decision-makers rated not only the importance of ten attributes to their choice, but also how different they perceived nursing homes to be on those attributes. When compared, determinant attributes were different from importance attributes. Respondents reported that cleanliness, location, and reputation were most determinant in nursing home choice. However, depending on profile characteristics such as income and relationship of the decision maker to the patient, determinant attributes differed. Therefore, profile characteristics influence determinant attributes. PMID- 11873458 TI - Facial asymmetry. The diagnostic challenge. AB - Evaluation of facial asymmetries requires a complete and systematic approach. This clinical examination is supplemented with radiographic studies including panoramic and cephalometric views. For complex cases CT scans are useful. Evaluation of the shape of the cervical vertebrae seen on lateral cephalometric radiographs can be used to predict potential growth in children. PMID- 11873459 TI - Reconstruction of secondary post-traumatic zygomatic and orbital deformities. PMID- 11873460 TI - Pathologic facial asymmetries. PMID- 11873462 TI - The use of autogenous fat grafts in the correction of facial asymmetries. AB - Successful long-term augmentation of facial contour defects can be anticipated provided the appropriate technique is used, careful patient selection is carried out, and meticulous atraumatic techniques are used both to harvest and to augment. PMID- 11873461 TI - Facial asymmetries in hemifacial microsomia, Goldenhar syndrome, and Treacher Collins syndrome. PMID- 11873463 TI - Secondary autogenous and alloplastic reshaping procedures for facial asymmetry. PMID- 11873464 TI - Management of facial asymmetry. Introduction. PMID- 11873465 TI - Nursing research and the human sciences. AB - Nursing has long been associated with the natural sciences. However, more recently some nurses have begun to identify it as a human science. The epistemological and ontological bases then shift, with clear implications for the research approaches that are regarded as the most useful. This column offers a discussion of the worldviews represented in contemporary nursing knowledge, focusing particularly on the newer paradigm that includes the view of nursing as a human science, and the place of qualitative and quantitative research. Neither the human sciences nor the natural sciences are seen as providing a sufficient base for all nursing knowledge. Both qualitative and quantitative approaches are viewed as potentially useful strategies given the nature of the phenomenon to be explored. Parallels to current discussions of worldviews and research methods in transpersonal psychology are identified. PMID- 11873466 TI - Ethical implications for acting faithfully in the nurse-person relationship. AB - Bearing witness-not bearing witness has been posited as a fundamental rhythm of coexistence. In biomedical ethical writings, the concept of fidelity, or faithfulness, has been used to describe an obligation to act in good faith with such actions as keeping promises or vows, fulfilling agreements, maintaining relationships, and individuals' fulfilling fiduciary responsibilities. In this column, the nursing phenomenon of acting faithfully in the nurse-person relationship shall be defined and explored with practice implications through reflections as viewed from the lens of the nursing theoretical framework of the human becoming school of thought. PMID- 11873468 TI - Indian health initiatives: a nursing practice model. PMID- 11873467 TI - Nursing science as human science: the new world and human becoming. AB - The human science perspective of the theory of human becoming holds intrinsic characteristics conducive to permeating teaching-learning in the 21st century. This column focuses on the challenges involved in the teaching-learning of nursing science as a human science, particularly from a human becoming perspective. Moving from a medical model of studying disease processes as the foundation for nursing education, to a nursing model of understanding the meaning of unitary lived experiences, poses challenges for both faculty and students in today's nursing world. These challenges include: explicating the unitary nature of humankind, living the responsibility of expressing new thought, perceiving teaching-learning as engaging community; and visioning nursing leadership as a transformative presence in change. PMID- 11873469 TI - A human science practice model for long-term care. PMID- 11873470 TI - The bridge: a model of human becoming practice. PMID- 11873472 TI - On science and human science: a conversation with Marilyn M. Rawnsley. Interview by Jacqueline Fawcett. PMID- 11873471 TI - Nursing knowledge and human science revisited: practical and political considerations. AB - The human science tradition is rooted in human freedom and meaning and oriented toward narrative and dialogical methods. In the past 10 years, human science nursing has grown but the opposition has also increased. Whereas other health disciplines are turning to the study of lived experience, nursing on the whole may be turning away. This article updates progress in human science, including works related to major nursing theories. The authors address practical and political considerations related to language, community, theory-laden knowledge, and tolerance for diversity. The authors conclude that the suppression of human science imperils nursing as a practice of being-with, witnessing, and cocreating quality of life, lived by nurses. But theories live in the actions of those who support them; thus, any place where people seek human care has the potential to support a human science-based nursing practice. PMID- 11873473 TI - Transforming healthcare with a unitary view of the human. AB - The purpose of this article is to explicate the meaning of unitary from a historical perspective. The term was first used in ancient Greece, and its meaning has been transformed over time. The article explicates the similarities and differences among the nurse theorists who use the term unitary. It also specifies a different approach to healthcare when a unitary perspective of the human guides practice. PMID- 11873474 TI - What is nursing science? AB - The enigma of defining nursing science is preceded by defining nursing, science, research, and nursing theory-guided practice. The context for exploring the meaning of nursing science is provided through examination of the totality and simultaneity paradigms. Differing views of nursing as a discipline are discussed. The position is taken that nursing is a basic science with various nursing schools of thought that constitute the substantive knowledge of the discipline. Finally, a definition of nursing science is presented that is broad enough to encompass all disciplinary knowledge. Despite current challenges, an optimistic vision is emerging. The nurse theorists and other nurse scholars who are furthering the development of this work are considered to be the cultural creatives of nursing and contributors to a larger movement toward wholeness in science and in society. PMID- 11873475 TI - Caring science in a new key. AB - A reorientation is going on in caring science. It could be called a new key characterized by more humanistically oriented thinking, which gives new significance to caring science. The sounding board of the new key is to be found in its ontological core. Its progress depends on whether we will succeed in laying bare the core of caring and developing its fundamental concepts and main theory. We need to regain the hermeneutical approach to penetrate into the core. Caring today needs this knowledge to help the patient in an increasingly complex world. PMID- 11873476 TI - Yes, no, or perhaps: reflections on Swedish human science nursing research development. AB - This article attempts to describe and analyze whether Swedish nursing research development belongs in a human science perspective. Traditionally, nursing in Sweden has been taught and practiced as a medical science, however another worldview seems to characterize Swedish nursing research today. This trend was identified through a study of the latest Swedish theoretical developments, in comparison with different international standpoints and views on the human science perspective. The conclusion is that Swedish nursing research is developing toward, and perhaps belongs in, a human science nursing perspective. PMID- 11873477 TI - Ambiguous opportunity: toiling for truth of nursing art and science. AB - This article questions traditional boundaries between nursing art and nursing science and explores how nurses build knowledge and truth. A brief overview of familiar notions about nursing art is followed by questions that are meant to deepen understanding about nursing and the knowledge required for a discipline. Authors describe understanding as an event that heralds human creation of meaning and potential action. Art is then shown to be a way to enhance understanding and meaningful knowledge when woven with nursing theory to guide practice. Findings from Parse's research method are described as artistic expressions, and borders that have served to separate notions about nursing art and science are challenged. The hermeneutics of human becoming are presented as beacons for truth and understanding. Authors call for tolerance of ambiguity and openness to support dialogue and discovery. PMID- 11873478 TI - Toward a global perspective of the human sciences. AB - Human science-based research, as defined by Dilthey and later developed by Gadamer, provides a valuable way to uncover what human beings have shared without denying important differences. It assumes that people are self-interpreting beings who seek meaning and purpose in the concrete situations of their lives. Gadamer's hermeneutic method is offered as an approach to do international and cross-cultural studies of relevance to nursing and healthcare. Three examples of human science-based nursing studies, which provide global understanding, are discussed. PMID- 11873479 TI - Household chores: under what conditions do mothers lean on daughters? PMID- 11873480 TI - Extended schooling, adolescence, and the renegotiation of responsibility among Italian immigrant families in New Haven, Connecticut, 1910-1940. PMID- 11873481 TI - Work contributions to the family: developing a conceptual and research framework. PMID- 11873482 TI - Family obligation and the academic motivation of adolescents from Asian, Latin American, and European backgrounds. PMID- 11873483 TI - Children investing in their families: the importance of child obligation in successful development. PMID- 11873484 TI - Biological dosimetry using human interphase peripheral blood lymphocytes. AB - Conventional metaphase-spread chromosome-aberration-based biodosimetry techniques for radiation dose assessment, although robust, are laborious and time consuming. The molecular cytogenetic laboratory of the Armed Forces Radiobiology Research Institute is developing simple and rapid interphase-based cytological assays that will be applicable to a broad range of radiation exposure scenarios. These assays include analysis of chromosome aberrations (premature chromosome condensation fluorescence in situ hybridization assay) and mitochondrial DNA mutations (mtDNA4977 deletion assay) using resting human peripheral blood lymphocytes. The dose-effect relationship for radiation-induced aberrations involving chromosome 1 after 24 hours of repair at 37 degrees C in resting human peripheral blood lymphocytes was studied using fluorescence in situ hybridization after chemical induction of premature chromosome condensation as previously explained. In the present study, we examined whether gamma irradiation in the range of 0 to 7.5 Gy induces a dose-dependent increase in aberrations manifested as "excess spots." The number of excess spots per cell, reflecting aberrations involving chromosome 1, increased from 0.035 at 0.5 Gy to 0.236 at 7.5 Gy. This observed dose-effect relationship was fit with a nonlinear power model. This technique may be extended to the study of radiation-induced translocations in interphase cells for retrospective dose reconstruction. With a recently developed in situ polymerase chain reaction method to detect and quantify mtDNA deletion in interphase cells after radiation exposure in cultured human peripheral blood lymphocytes, 90% to 95% of cells are analyzable. We discuss the potential use of the mtDNA deletion assay in biological dosimetry applications. Interphase-based cytological assays may eliminate some inherent problems associated with metaphase-spread-based assays. These problems involve (1) the limited number of analyzable cells containing chromosome aberrations, which is due to various factors including radiation-induced cell death and delay in cell cycle progression into mitosis, and (2) the requirements for radiation cytogenetics expertise and tedious labor to manually score chromosome aberrations. PMID- 11873485 TI - Sulfur mustard medical countermeasures in a nuclear environment. AB - The possibility of chemical warfare occurring on a nuclear battlefield exists, given the increased proliferation of both types of weapons in small nations during the last two decades. Antidotes to mustard "gas" have not yet been fielded because its mechanism of action is not fully understood. Researchers at the U.S. Army Medical Research Institute of Chemical Defense have been addressing this dilemma and have developed six intervention strategies that may have efficacy for both chemical and nuclear weapons. These strategies include intracellular scavengers, DNA cell cycle modulators, poly(ADP-ribose) polymerase inhibitors, calcium modulators, anti-proteases, and anti-inflammatories. Because mustard and radiation may produce synergistic effects that could impair the efficacy of individual therapies, studies have been proposed to evaluate the combined effects of nuclear and chemical exposures. Once models for these studies have been established, the safety and efficacy of the intervention strategies can be evaluated. PMID- 11873486 TI - Bacillus anthracis infection in irradiated mice: susceptibility, protection, and therapy. PMID- 11873487 TI - Management of postirradiation sepsis. AB - Ionizing radiation depresses the immune defenses and enhances susceptibility to local and systemic infection due to endogenous or exogenous microorganisms. Exposure to a lethal dose of ionizing cobalt-60 gamma radiation induces a dose related reduction in the number of both aerobic and anaerobic bacteria from 10(10 12) to 10(4-6) per gram of stool within 4 days. Whereas the number of anaerobic bacteria stays low, the number of Enterobacteriaceae per gram of stool increases significantly--up to 10(9) by the 12th day following irradiation. This increase is associated with bacterial translocation of these organisms and fatal bacteremia. The use of quinolones was effective in controlling systemic endogenous Gram-negative infection following irradiation. Supplementation with penicillin prevented treatment failures due to Streptococci and increased survival. Quinolones given for 21 days were also effective in the management of systemic exogenous infections due to orally ingested Klebsiella pneumoniae and Pseudomonas aeruginosa. Quinolones may be effective because they inhibit growth of the exogenous organism within the gut lumen while preserving the anaerobic gut flora and their systemic antibacterial activity. Coadministration of antimicrobials effective against anaerobes may be required for the management of polymicrobial infections. The availability of both oral and parenteral routes of administration, the advantage of achieving selective inhibition of potential pathogens in the gut, and the ability to treat systemic infection make the quinolones promising agents for the therapy of endogenous and exogenous infections after irradiation. PMID- 11873488 TI - Combined effects modeling of ionizing radiation and biological agent exposures. AB - A simple two-species population model is used to simulate the interactions of a replicating pathogen and the immune system response. This deterministic model is flexible enough to reproduce the salient population-dynamic features, chiefly the tendency toward an equilibrium state in which the pathogen population is reduced to zero or a negligible level after some finite time. The specific immune system species that can be taken initially at zero increases to a finite equilibrium level that represents an immune system "memory" of a given pathogen. The coefficients in the model equations represent production, interaction, and loss rates. Their relative sizes determine the qualitative behavior of the solution in its phase space. The effect on the immune system of exposure to ionizing radiation is modeled by reducing the production rate coefficients; the reduction is proportional to the level of ionizing radiation. According to the model and based on historical hematopoietic data from retrospective accident studies, exposures to ionizing radiation can cause pathogen levels to rise an order of magnitude or more than the baseline (no radiation exposure) case and, for some simulated levels, escape from the bounded region of the phase space. A means is offered to link the deterministic model of time-varying pathogen level in an individual host to the corresponding probability of lethality (increasing with time) of an individual in a population. PMID- 11873489 TI - Management of cutaneous radiation injuries: diagnostic and therapeutic principles of the cutaneous radiation syndrome. AB - The cutaneous symptoms that appear after radiation exposure are caused by a combination of inflammatory processes and alteration of cellular proliferation as a result of a specific pattern of transcriptionally activated proinflammatory cytokines and growth factors. The symptoms follow a time course consisting of prodromal erythema, manifestation, chronic stage, and late stage; these symptoms are referred to as cutaneous radiation syndrome (CRS). The time course depends on several factors such as the applied radiation dose, radiation quality, individual radiation sensitivity, extent of contamination and absorption, and volume of skin exposed. For diagnosis of CRS, the following procedures are used: 7.5 to 20 MHz B scan sonography, thermography, capillary microscopy, profilometry, nuclear magnetic resonance imaging, bone scintigraphy, and histology. Based on the results of previous experimental and clinical research, pharmacotherapy of CRS can include topical or systemic application of corticosteroids, gamma interferon, pentoxifylline and vitamin E, and superoxide dismutase. The treatment varies according to the stage of CRS. Due to the complexity of the clinical manifestations of radiation disease in most patients, interdisciplinary treatment at specialized centers is necessary. In most cases, dermatologists are asked to provide lifelong therapy and follow-up of the patients. PMID- 11873490 TI - Nuclear/biological/chemical combined injury effects: expert panel consensus. PMID- 11873491 TI - Health effects of embedded depleted uranium. AB - The health effects of embedded fragments of depleted uranium (DU) are being investigated to determine whether current surgical fragment-removal policies are appropriate for this metal. The authors studied rodents implanted with DU pellets as well as cultured human cells exposed to DU compounds. Results indicate that uranium from implanted DU fragments distributes to tissues distant from implantation sites, including bone, kidney, muscle, and liver. Despite levels of uranium in kidney that would be nephrotoxic after acute exposure, no histological or functional kidney toxicity was observed with embedded DU, indicating that the kidney adapts when exposed chronically. Nonetheless, further studies of the long term health impact are needed. DU is mutagenic and transforms human osteoblastic cells into a tumorigenic phenotype. It alters neurophysiological parameters in rat hippocampus, crosses the placental barrier, and enters fetal tissue. Preliminary data also indicate decreased rodent litter size when animals are bred 6 months or longer after DU implantation. PMID- 11873492 TI - Potential late health effects of depleted uranium and tungsten used in armor piercing munitions: comparison of neoplastic transformation and genotoxicity with the known carcinogen nickel. AB - Limited data exist to permit an accurate assessment of risks for carcinogenesis and mutagenesis from embedded fragments or inhaled particulates of depleted uranium (DU). Ongoing studies have been designed to provide information about the carcinogenic potential of DU using in vitro and in vivo assessments of morphological transformation as well as cytogenetic, mutagenic, and oncogenic effects. For comparison, we also examined tungsten alloys used in military projectiles and the known carcinogen nickel. Quantitative and qualitative in vitro transformation studies were done to assess the carcinogenic potential of radiation and chemical hazards. Using a human osteosarcoma cell model, we demonstrated that soluble and insoluble DU compounds can transform cells to the tumorigenic phenotype, as characterized by morphological, biochemical, and oncogenic changes consistent with tumor cell behavior. Tungsten alloys and nickel were also shown to be neoplastic transforming agents, although at a frequency less than that of DU. Sister chromatid exchange, micronuclei, and alkaline filter elution assays showed DU and tungsten alloys were genotoxic. Exposure to a nontoxic, nontransforming dose of DU induced a small but statistically significant increase in the number of dicentrics formed in cells. These results suggest that long-term exposure to DU or tungsten alloys could be critical to the development of neoplastic disease in humans and that additional studies are needed. PMID- 11873493 TI - Health effects and biological monitoring results of Gulf War veterans exposed to depleted uranium. AB - A small group of Gulf War veterans have retained fragments of depleted uranium (DU) shrapnel, the long-term health consequences of which are undetermined. We evaluated the clinical health effects of DU exposure in Gulf War veterans compared with nonexposed Gulf War veterans. History and follow-up medical examinations were performed on 29 exposed veterans and 38 nonexposed veterans. Outcome measures used were urinary uranium determinations, clinical laboratory values, and psychiatric and neurocognitive assessment. Gulf War veterans with retained DU metal shrapnel fragments were found to be still excreting elevated levels of urinary uranium 7 years after first exposure to DU (range for exposed individuals is 0.01-30.7 micrograms/g creatinine vs. 0.01-0.05 microgram/g creatinine in the nonexposed). The persistence of the elevated urine uranium suggests ongoing mobilization of uranium from a storage depot, resulting in chronic systemic exposure. Adverse effects in the kidney, a presumed target organ, were not seen at the time of the study; however, other subtle effects were observed in the reproductive and central nervous systems of the DU-exposed veterans. PMID- 11873494 TI - Depleted uranium: a radiochemical toxicant? AB - The first large-scale combat use of depleted uranium (DU) weapons occurred during the Gulf War, and some U.S. personnel were wounded by DU fragments. Established fragment removal policies dictated that embedded metal fragments be left in place unless doing so posed unacceptable additional risks. However, questions were raised as to whether these policies are appropriate for a metal that--unlike lead, steel, or others--is chemically toxic and emits low-level radiation. Data from research currently under way indicate that long-term exposure to embedded DU fragments may present a level of risk that requires modification of established policies. Our understanding of DU health effects and of the possible mechanisms by which DU might affect tissues is evolving. Understanding more about the long term response of tissues exposed to DU could facilitate future development of treatments for DU injuries. PMID- 11873495 TI - Radiation-induced performance decrement. AB - This article is a brief review of performance decrement expected after low-level radiation doses (0.70 Gy or less) and after higher radiation doses. Examples are presented from both animal and human data. The data indicate that low-level radiation doses in humans are not expected to degrade performance on the battlefield. Doses higher than 1.25 Gy are likely to degrade performance. Transient performance deficits known as early transient incapacitation observed in animals and humans after large, rapidly delivered doses of ionizing radiation are discussed. PMID- 11873496 TI - Stress-gene induction by low-dose gamma irradiation. AB - Using a human myeloid tumor cell line (ML-1), we detected induction of mRNA expression of several stress-responsive genes by doses of gamma rays as low as 2 cGy. For instance, the dose response for induction of CIP1/WAF1 and GADD45 appears to be linear over the range of 2 to 50 cGy and shows no evidence of a threshold for induction. Although 2 and 5 cGy exposures did not result in any detectable reduction in cloning efficiency nor in increased apoptosis in ML-1 cells, these exposures did produce a brief cell-cycle delay. We also used fluorescent cDNA microarray hybridization to investigate transcriptional stress responses following low doses of gamma rays and to identify additional radiation responsive genes for inclusion in a stress-specific microarray we are developing. These studies provide insight into the molecular responses to physiologically relevant doses, which cannot necessarily be extrapolated from high-dose studies. The use of high throughput arrays will allow the identification of multiple stress-responsive genes that are radiation inducible in a variety of cell types and tissues. The expectation is that transcriptional stress responses will provide a molecular approach to monitoring for radiation exposure and detecting interindividual differences. PMID- 11873497 TI - Placing long-term and acute radiation effects in a common framework. AB - No method presently exists for placing long-term risks from low-level radiation doses and acute radiation sickness (ARS) from high-level doses within a common framework useful for the military commander. This article proposes such a method. The Defense Threat Reduction Agency and its predecessor agencies pioneered the quantitative description of the impact of ARS on human performance capability and military unit effectiveness. One product of these efforts is the Radiation Induced Performance Decrement (RIPD) software. RIPD software includes physiologically based models of ARS valid for both chronic and one-time radiation exposure. One key element of placing ARS and low-level radiation effects in a common framework is finding independent and dependent variables that have a common meaning for both. This paper examines the use of cumulative probability of effect vs. dose and time after exposure for establishing a common framework. The physiologically based models of upper gastrointestinal distress and fatigability and weakness from the RIPD software support this approach. PMID- 11873498 TI - Radiophobia: long-term psychological consequences of Chernobyl. AB - The primary health effect of Chernobyl has been widespread psychological distress in liquidators (workers brought in for cleanup), evacuees, residents of contaminated areas, and residents of adjacent noncontaminated areas. Several psychoneurological syndromes characterized by multiple unexplained physical symptoms including fatigue, sleep and mood disturbances, impaired memory and concentration, and muscle and/or joint pain have been reported in the Russian literature. These syndromes, which resemble chronic fatigue syndrome and fibromyalgia, are probably not due to direct effects of radiation because they do not appear to be dose related to radiation exposure and because they occur in areas of both high and low contamination. PMID- 11873499 TI - Human responses to the threat of or exposure to ionizing radiation at Three Mile Island, Pennsylvania, and Goiania, Brazil. AB - The psychological stressors and their aftereffects associated with the Three Mile Island accident, the Goiania, Brazil, cesium-137 accident, and the Abadia, Brazil, storage location are summarized and compared. Cross-cultural comparisons of human responses to ionizing radiation are rare. A multidisciplinary methodological approach to examining the psychological responses to ionizing radiation is even more rare. The psychological, behavioral, neuroendocrine, and cardiovascular results are summarized for Three Mile Island, Goiania, and Abadia. PMID- 11873500 TI - Psychological consequences of military operations in low-level radiation environments. AB - Military operations have produced neuropsychiatric (NP) casualties throughout history. The same is true for radiation accidents. Although we have not carried out military operations in nuclear environments, related experience suggests that operations performed under low-level radiation conditions could serve to compound the factors that are known to produce NP casualties. Historically, military NP casualty rates have varied widely depending on a complex array of factors, ranging from the actual conditions of the operation and experiences of the soldiers to the perception of the conflict by both the soldiers and the people back home. History has also shown that lack of preparation contributes to NP casualties. The number and severity of NP casualties can be minimized by ensuring that the equipment, training, and leadership are in place to instill in our soldiers the confidence that they can cope with the types of threats that are part of modern-day missions. PMID- 11873501 TI - Physiological and psychological impact of low-level radiation: an overview. PMID- 11873502 TI - Nucleic acid molecular biomarkers for diagnostic biodosimetry applications: use of the fluorogenic 5'-nuclease polymerase chain reaction assay. AB - A reliable, relatively easy method for diagnostic assessment of radiation exposure is needed to support the triage of radiation casualties and medical treatment decisions in military defense operations. Our strategy is to identify radiation-responsive DNA mutations and gene expression targets that can be analyzed using polymerase chain reaction (PCR) assays and an existing fluorescence-based nucleic acid analysis system designed for forward-deployable laboratory applications. Using an in vitro model system of human peripheral blood lymphocytes, we identified a candidate nucleic acid biomarker (i.e., gene expression target) that is responsive to ionizing radiation. In this report, we describe our preliminary Haras gene expression findings. A dose-dependent elevation in Haras gene expression levels was demonstrated using Northern-blot analysis 17 hours after exposure to a 250-kVp dose of X-rays (25-100 cGy, 1 Gy/minute); c-Haras expression levels at 100 cGy were ninefold higher than those of controls. An alternative protocol to quantify the Haras cDNA target, using the rapid, real-time reverse transcriptase fluorogenic 5'-nuclease PCR assay, is described, along with a preliminary characterization of the dynamic range for detection. Our research shows that the analysis of multitarget nucleic acid biomarkers, using the multiplex fluorogenic 5'-nuclease PCR assay, has beneficial applications in radiation epidemiology, radiation therapy, and biodosimetry. PMID- 11873503 TI - Immunofluorescence detection of radiation-induced DNA base damage. AB - We previously described a sensitive assay for measuring thymine glycol in the DNA of irradiated cells. The assay combines immunorecognition of the DNA lesion with capillary electrophoresis and laser-fluorescence detection to achieve an absolute detection level in the zeptomole (10(-21) mol) range. This article describes modifications to the protocol that overcome certain technical problems seen with the original methodology. In particular, the capillary electrophoresis is carried out at pH 8.3 rather than pH 10.5. The new protocol was used to examine removal of thymine glycol from the DNA of A549 lung adenocarcinoma cells and resting lymphocytes after exposure to 2 Gy gamma rays. Both cell types displayed similar repair kinetics. Removal of thymine glycol is almost complete at 4 hours postirradiation. PMID- 11873505 TI - Environmental monitoring technology. AB - As a consequence of the new geopolitical environment, missions of military forces have changed. Soldiers are now primarily involved in peacekeeping or humanitarian missions. Even the radiological threat has changed. Now there is greater concern about exposure to low-level radiation (LLR) as opposed to that from thermonuclear warfare. The North Atlantic Treaty Organization (NATO) defines LLR as radiation deriving from all but nuclear weapon sources, regardless of the intensity of the radiation. LLR raises new questions about physical dosimetry requirements of military forces in managing radiological situations. After presenting a list of potential radiological threats, this paper discusses questions raised by LLR, defines problems in operational terms, and suggests possible solutions. Finally, the new French military dosimeter is presented as the first attempt to create a military dosimeter useful for both peacetime and wartime. PMID- 11873507 TI - A survey of physical dosimetry to date and in the near future: Part 1. Review of standards and regulatory issues. AB - This article summarizes the status of the relevant standards and current regulatory issues for use of physical dosimetry devices for the occupational worker in the United States. Included is a summary of relevant standards from the International Organization for Standardization (ISO), the International Electrotechnical Commission (IEC), the American National Standards Institute (ANSI), the United States Nuclear Regulatory Commission NUREG-Series, the National Voluntary Laboratory Accreditation Program (NVLAP), the Department of Energy Laboratory Accreditation Program (DOELAP), and the U.S. Military Specifications and Standards (MIL-STD). Proposed changes to ANSI N13.11-1993, "American National Standard for Dosimetry-Personnel Dosimetry Performance Criteria for Testing," are listed. The strategic changes that the United States Nuclear Regulatory Commission (NRC) is making in rulemaking activities related to dosimetry and standards are given. The status of Measurement Program Description (MPD) C.18, "Implementation of Electronic Dosimetry for Primary Dosimetry," from the Council on Ionizing Radiation Measurements and Standards (CIRMS) is given. PMID- 11873504 TI - Overview of low-level radiation exposure assessment: biodosimetry. AB - The capability to make diagnostic assessments of radiation exposure is needed to support triage of radiation casualties and medical treatment decisions in military operations. At the International Conference on Low-Level Radiation Injury and Medical Countermeasures session on biodosimetry in the military, participants reviewed the field of biomarkers, covering a wide range of biological endpoints. Participants evaluated early changes associated with exposure to ionizing radiation, including chromosomal and DNA damage, gene expression and associated proteins, and DNA mutations. The use and development of advanced monitoring and diagnostic technologies compatible with military operations was emphasized. Conventional radiation bioassays require a substantial amount of time between when the sample is taken and when the data can be provided for decision making. These "reach back" bioassays are evaluated in laboratories that are not in the field; these laboratories routinely measure exposures of 25 cGy (photon equivalent levels). Detection thresholds can be reduced approximately fivefold by the addition of significant and tiresome scoring efforts. Alternative real-time biomarkers that can be measured in field laboratories or with handheld detection devices show promise as screening and clinical diagnostic tools, but they require further development and validation studies. PMID- 11873508 TI - A survey of physical dosimetry to date and in the near future: Part 2. Review of commercially available products. AB - This article summarizes the status of physical dosimetry for the occupational worker. The review of commercially available physical dosimetry systems was limited to the following technologies: thermoluminescent dosimeters, electronic personnel dosimeters, optically stimulated luminescence dosimeters, and direction ion storage dosimeters. Product reviews were limited to the top models and largest commercial manufacturers in each category. The physical principles of each dosimeter type are discussed. Information was gathered from journal literature, by direct experience, and by inviting six commercial vendors to present their newest technologies. Each system was found to have strengths and weaknesses. Many of the technologies presented by major vendors were still in development and thus could be considered near-future systems. PMID- 11873509 TI - Absorbed dose and the quantification of physical events at low-dose irradiation of tissue. PMID- 11873510 TI - Consensus regarding low-level radiation assessments and countermeasures: physical dosimetry. AB - One of the goals of the 1999 International Conference on Low-Level Radiation Injury and Medical Countermeasures was to review current and anticipated capabilities for measuring the quantity and quality of radiation exposures using personal dosimetry systems, fieldable spectroscopy equipment, and modeling strategies. Technological advances in these systems need to be evaluated for use in radiation exposure assessment. The conference focused on the application of these systems to military operations, emphasizing their use for radiation exposure assessment internationally. Capability exists to provide precise assessment of external dose during military operations, but consideration must be given to the deployability and practicality of providing additional gear for military personnel. Assessment of internal dose in the field continues to be a problem, as there are no current methods to rapidly assess intake or to account for resuspension of radioactive material due to the operation itself. Modeling can assist in overcoming confounding factors such as source-dependent energy deposition and decay, shielding, and radioisotope distribution. PMID- 11873511 TI - Sensitivity to low-level radiation in radiosensitive "wasted" mice. AB - Mice homozygous for the autosomal recessive "wasted" mutation (wst/wst) have abnormalities in T lymphocytes and in the anterior motor neuron cells of the spinal cord, leading to sensitivity to low doses of ionizing radiation, hind limb paralysis, and immunodeficiency. This defect results in a failure to gain weight by 20 days of age and death by 28 days. The wasted mutation (previously mapped to mouse chromosome 2) is shown to be a 3-bp deletion in a T cell-specific (and perhaps motor-neuron-specific) regulatory region (promoter) of the proliferating cell nuclear antigen (PCNA) gene on mouse chromosome 2. A regulatory element is also shown to be important in PCNA expression in T lymphocytes and motor neuron cells affected by the 3-bp deletions in the PCNA promoter. The model is as follows. Absence of PCNA expression in the thymuses (and motor neurons) of wasted mice causes cellular apoptosis. This absence of expression is mediated by a positive transfactor that can bind to the wild-type but not to the wasted mutant PCNA promoter. The bound protein induces late expression of PCNA in T lymphocytes and prevents onset of radiation sensitivity in the cells. PMID- 11873512 TI - Genomic instability and bystander effects: a paradigm shift in radiation biology? AB - A basic paradigm in radiobiology is that, following exposure to ionizing radiation, the deposition of energy in the cell nucleus and the resulting damage to DNA, the principal target, are responsible for the radiation's deleterious biological effects. Findings in two rapidly expanding fields of research- radiation-induced genomic instability and bystander effects--have caused us to reevaluate these central tenets. In this article, the potential influence of induced genomic instability and bystander effects on cellular injury after exposure to low-level radiation will be reviewed. PMID- 11873513 TI - Hematopoietic cell renewal as the limiting factor in low-level radiation exposure: diagnostic implications and therapeutic options. AB - In some radiation accidents, exposure doses are delivered over days or even months. In all cases the organ system most relevant to a patient's survival is the hematopoietic tissue. There appears to be a threshold of approximately 10 mSv per day above which hematopoietic effects become apparent and hematopoietic failure may occur. Experimental observations in dogs demonstrate that exposure to chronic gamma-irradiation may be tolerated for over a year if the daily dose does not exceed 7 mSv to 18 mSv. The pathophysiological mechanisms are being studied by hematological measures and biomathematical models. The results are in accordance with the assumption of excess cell loss and progressive diminution of the stem cell pool over time until a "turbulence region," with an increased probability of system failure, is approached. Diagnostic procedures require a thorough hematological assessment that includes the stem cell compartment. Therapeutic options include administration of hematopoietic growth factors and stem cell transplantation. PMID- 11873514 TI - Nitroxides as radiation protectors. AB - Selective protection of normal tissues from the damaging effects of ionizing radiation is an important objective in cancer treatment research. Likewise, radioprotective agents may be useful in protecting the human population in the event of radiation-related accidents or military conflicts. Over the past decade, we have identified stable nitroxide compounds as a unique class of antioxidants with demonstrated radioprotective properties. We have shown that nitroxides at nontoxic concentrations are effective as in vitro and in vivo antioxidants when oxidation is induced by superoxide, hydrogen peroxide, organic hydroperoxides, ionizing radiation, or specific DNA-damaging anticancer agents. Studies have shown that nitroxides protect against radiation-induced DNA damage, chromosome aberrations, mutation induction, cell killing, and lethality in mice that receive whole-body irradiation. Whether these agents provide radioprotection for low level radiation doses remains to be determined. PMID- 11873515 TI - Detection of single-strand breaks and base damage in DNA of human white blood cells as a tool for biological dosimetry of exposure to ionizing radiation. AB - An immunochemical assay has been adapted to detect DNA damage in whole blood at biologically relevant doses of ionizing radiation. Upon alkaline treatment of whole blood, both strand breaks and base damage (which is converted into strand breaks by the addition of damage-specific enzymes) are detected by using antibodies that specifically bind to single-strand DNA. Single-strand breaks can be detected immediately after irradiation at doses as low as 0.2 Gy. With unknown background damage, the lower detection limit increased to approximately 0.5 Gy immediately after irradiation due to interindividual variation. Because single strand breaks are repaired rapidly, this method is suitable only for blood collected less than 1 hour after exposure. Base damage represents a very promising biological indicator that can be used 1 hour and longer (at least to 4 hours) after radiation exposure because of an apparent lack of base damage repair during this time window. PMID- 11873516 TI - Relationships between cytoprotection and mutation prevention by WR-1065. AB - WR-2721, the leading compound that has emerged from the U.S. Army's Drug Synthesis and Development Program, has become the first clinically used cytoprotector in cancer therapy. The ability of its active thiol, WR-1065, to act as an antioxidant to scavenge free radicals, donate hydrogen atoms, and induce auto-oxidation is related to its cytoprotective effectiveness. However, at concentrations 10 to 20 times lower than those required for cytoprotection, this nonprotein thiol also affects gene expression, transcription factor binding, protein phosphorylation levels, and mutagenesis. For example, WR-1065 treatment activates binding of the redox-sensitive transcription factor NF-kappa B to DNA and affects expression of a number of genes, including thymidine kinase, c-myc, and manganese superoxide dismutase. At these lower drug concentrations, phosphorylation levels and subsequent activities on intracellular enzymes are also inhibited. These observations suggest that the mechanisms of action of WR 2721 and its metabolites to protect against cell killing are different from those that protect against mutagenesis and cancer formation. The ability to affect these postexposure processes makes WR-2721 an important agent in the reduction of the carcinogenic risks of environmental exposure to radiation or deleterious chemicals. PMID- 11873517 TI - Development of chemopreventive strategies for radiation-induced cancer: targeting radiation-induced genetic alterations. AB - Carcinogenesis is a multistage process involving dysregulation of signal transduction and cell cycle pathways. This dysregulation results in specific molecular and genetic alterations, including gene amplification, mutations, and chromosomal rearrangements. These aberrations can be measured to provide a novel means to assess carcinogenic risk or as targets for chemointervention. Recent human and in vivo studies have demonstrated that genetic alterations, such as oncogenes and oncoproteins, were observed in preneoplastic tissues or serum following exposure to chemical carcinogens or low-level radiation (LLR). Identification of preneoplastic changes following radiation exposure may provide information that will allow development of LLR chemopreventive strategies. Radiation carcinogenesis studies in vivo with a lung tumor model showed that a low-dose cobalt-60 radiation exposure induced persistent time-dependent genetic alterations, such as elevated ras expression. This radiation exposure also resulted in lung tumor formation in 26% of the irradiated animals at 232 days after irradiation. A significant and progressive increase in ras oncogene expression was measured using Northern blot analysis in 80% of the irradiated animals over the duration of the experiment. Pharmacological intervention strategies are being tested using buthionine-[S,R]-sulfoximine (BSO). BSO has been previously shown to down-regulate ras expression. Administration of BSO prevented radiation-induced changes in ras mRNA levels in this lung tumor model. Further studies are being conducted with an LLR-induced leukemia model in which detection of circulating levels of oncoproteins will be more feasible. Based on these preliminary results and on its clinical efficacy and low clinical toxicity, BSO warrants further study as an LLR chemopreventive agent. Furthermore, this strategy to target LLR-induced preneoplastic alterations may be an effective means of developing modulators of LLR-induced cancers. PMID- 11873518 TI - Nutritional approaches to radioprotection: vitamin E. AB - Low-level radiation injury is dependent on the radiation dose and dose rate. The major military use of any potential radioprotectant is to prevent the short-term effects of lethality and the long-term effects of cancer and other pathologies from radiation exposure that may occur in a nuclear battlefield or in a nuclear material contaminated field of operation. Therefore, a radioprotectant should not affect the ability of military personnel to perform tasks. Because exposure to ionizing radiation induces free radical species, effective antioxidants, either alone or in combination with other agents, can be used as potential radioprotectors. To test this hypothesis, we studied vitamin E for its radioprotective efficacy. Using CD2F1 male mice as the model system, we observed that vitamin E at a dose of 400 IU/kg acts as a good radioprotectant against lethal doses of cobalt-60 radiation. Vitamin E was more efficacious when given subcutaneously than when given orally. PMID- 11873519 TI - Inhibition of adjuvant-induced arthritis by 16 alpha-fluoro-5-androsten-17-one. AB - The adrenal steroid dehydroepiandrosterone (DHEA) produces cancer-preventive, antiatherosclerotic, antidiabetic, immunomodulating, and anti-inflammatory effects in laboratory animals. The clinical use of DHEA is limited by its androgenicity. We have developed a synthetic congener of DHEA called fluasterone that, in animal tests, lacks the androgenicity, estrogenicity, and peroxisome proliferating effects of DHEA but retains the cancer-preventive, antidiabetic, and anti-inflammatory efficacy. This report discusses how fluasterone ameliorates the development of joint inflammation in an adjuvant-arthritis model in Lewis rats. PMID- 11873520 TI - Protection against gamma-irradiation with 5-androstenediol. AB - We showed previously that treatment of gamma-irradiated female B6D2F1 mice with 5 androstenediol (AED) enhanced survival, stimulated myelopoiesis, and ameliorated radiation-induced decreases in circulating neutrophils and platelets. We have now tested survival in male CD2F1 mice, and we have investigated molecular and functional effects on neutrophils and bone marrow stromal cells and screened for toxicity in female B6D2F1 mice. AED (160 mg/kg, subcutaneously, 24 hours before irradiation) enhanced survival in male CD2F1 mice with a dose-reduction factor of 1.23, similar to the dose-reduction factor of 1.26 found previously for female B6D2F1 mice. Expression of CD11b, a developmental marker, was reduced on circulating neutrophils after either in vivo AED administration or whole-body gamma-irradiation (3 Gy), but neutrophil peroxidase activity was unchanged. Stromal cell progenitors (fibroblastoid colony-forming units) were reduced in marrow 5 days after AED injection in nonirradiated mice. Clinical chemistry, histopathology, and behavioral assays showed no evidence of toxicity. We conclude that AED and related steroids are attractive candidates to explore as countermeasures to high- and low-level ionizing radiation. PMID- 11873521 TI - NF-kappa B, cytokines, proteasomes, and low-dose radiation exposure. AB - Ionizing radiation shares with proinflammatory cytokines a pathway that involves reactive oxygen species and activation of the redox-sensitive nuclear transcription factor NF-kappa B, which leads to expression of inflammatory and cell survival programs. NF-kappa B activation normally requires phosphorylation of its inhibitor I kappa B and the inhibitor's subsequent degradation by the proteasome. Nonlinear dose-response curves have been reported for both radiation induced cytokines and NF-kappa B and I kappa B expression with maximum exposures of less than 2 Gy and greater than 4 Gy, respectively. Radiation-inhibited proteasomes function over a wide dose range, suggesting that the proteasome is a redox-sensitive target for radiation that may function along with transcription to cause nonlinear dose-response relationships for early expression of many molecules, including NF-kappa B and cytokines. These pathways are relevant to low dose radiation effects, adaptive responses, and carcinogenesis. PMID- 11873522 TI - Cytokine-based treatment of radiation injury: potential benefits after low-level radiation exposure. AB - The use of growth factors (GFs) in the treatment of radiation injury has focused on enhancing recovery from acute radiation syndrome. A number of new GFs have shown significant in vivo and in vitro preclinical efficacy; some of these have recently been approved by the Food and Drug Administration, some are in various phases of clinical trials, and some are moving through preclinical evaluations. The most promising new GFs in the context of enhancing the viability of irradiated hematopoietic stem cells (HSCs) are flt-3L, c-kitL, and c-mplL. These GFs, as well as interleukin 3 (IL-3), have been shown to maintain viability, suppress apoptosis, and promote the clonal growth of primitive murine and human hematopoietic progenitor cells. Further evidence suggests that these GFs may also act in synergy with each other. Additionally, three families of chimeric proteins that consist of dual GF receptor (R) agonists have been engineered: myelopoietin, promegapoietin, and progenipoietin. These proteins activate the IL-3 and granulocyte colony-stimulating factor Rs, the IL-3 and mpl Rs, and the flt-3L and granulocyte colony-stimulating factor Rs, respectively. The preclinical data indicate that the chimeric GFR agonists are potent stimulators of hematopoiesis in myelosuppressed nonhuman primates and can effectively alleviate acute radiation syndrome in animals. Acute or protracted low-level radiation exposure does not require the extensive clinical care necessary following radiation induced myelosuppression. The main question is whether these new GFs will allow for enhanced repair of radiation-induced chromosome aberrations while promoting early survival of HSCs. Other questions include the following: Will an early, brief exposure to GFs suppress p53-dependent apoptosis and induce expression of bcl-2 with a concomitant enhancement of DNA repair capacity? What is the effect of GF stimulation of irradiated HSCs on p53 cell cycle checkpoint activity? Will GFs promote survival of "transformed" cells that would otherwise be eliminated by p53 activation of apoptosis-promoting genes? Relevant animal models and access to appropriate GFs will be required to answer these questions. PMID- 11873523 TI - Radioprotection of lung and esophagus by overexpression of the human manganese superoxide dismutase transgene. AB - Protection of normal tissue from radiation-induced damage has been demonstrated in the murine lung and esophagus using human manganese superoxide dismutase (MnSOD) plasmid/liposome complex (PL). C57BL/6J mice were injected intratracheally with MnSOD-PL, LacZ-PL, or metallothionein (MT2)-PL and irradiated 24 hours later at 2,000 cGy to the pulmonary cavity. Mice injected with MnSOD-PL had significantly increased survival compared with control, LacZ PL, or MT2-PL irradiated mice. In an esophagitis model, male C3H/HeJ mice were injected intraesophageally with either MnSOD-PL or LacZ-PL and irradiated 24 hours later at 3,500 cGy to the pulmonary cavity. Mice injected with MnSOD-PL had significantly increased survival after irradiation at 3,500 cGy compared with control or LacZ-PL injected mice. However, orthotopic 3LL lung tumors in C57BL/6J mice were not protected from radiation following injection of MnSOD-PL, as seen by increased survival of mice with these tumors following irradiation. MnSOD-PL gene therapy protected normal lung and esophagus but not 3LL orthotopic lung tumors from radiation-induced damage. PMID- 11873524 TI - Ex vivo expansion of bone marrow and cord blood cells to produce stem and progenitor cells for hematopoietic reconstitution. AB - Ex vivo expansion of a small volume of bone marrow offers an alternate approach to cellular support for repair of a hematopoietic system damaged by radiation exposure. Unpurified bone marrow cells cultured using frequent exchange of medium (perfusion) results in (1) the growth of an adherent layer that provides a supportive microenvironment for hematopoiesis and (2) the growth of hematopoietic stem and progenitor cells. Similar increases in stem and progenitor cells were also observed in cord blood cultures using frequent medium exchange methods. An automated clinical scale system, the AastromReplicell Cell Production System, has been developed to implement the biology of this ex vivo expansion process. Clinical use of ex vivo expanded bone marrow alone or in augmenting a low dose of mobilized peripheral blood stem cells has successfully reconstituted hematopoiesis. Ex vivo expanded cord blood cells combined with unmanipulated cord blood cells have also shown significant benefits in overall survival and engraftment in pediatric and adult patients, respectively. These results suggest that ex vivo expansion of hematopoietic cells provides a mechanism for generating cells capable of hematopoietic reconstitution. PMID- 11873525 TI - Sensitivity, specificity, and persistence of chromosome translocations for radiation biodosimetry. AB - The analysis of chromosome translocations by fluorescence in situ hybridization ("whole chromosome painting") is the gold standard endpoint for radiation biological dosimetry. Translocation formation requires mis-repair of two or more DNA double-strand breaks, a critical class of damage that is efficiently induced by ionizing radiation in a dose-responsive manner. Translocations are widely used for radiation exposure assessment because they provide a sensitive detection system for low doses, show good specificity for radiation exposure, persist for decades, and are highly relevant to tumorigenesis. This paper provides a brief evaluation of the use of translocation analyses by chromosome painting for biological dosimetry following chronic or low-dose exposure to ionizing radiation. PMID- 11873526 TI - Accommodative responses to chronic irradiation: effects of dose, dose rate, and pharmacological response modifiers. AB - Low-level radiation exposures are expected to have long-term health implications but few near-term effects that would impair function. These assumptions are based on extrapolation from acute exposure responses, not on studies of a larger array of exposure scenarios (e.g., protracted exposures) that might present as operational threats. Protracted exposure is one scenario that needs to be better defined in terms of both the initial effect and the long-term health consequences. Reported here are the near-term effects of chronic, low daily-dose gamma-irradiation (3-128 mGy per day) on the blood-forming system of canines. Change in hematopoietic capacity was monitored along with time of exposure and cumulative radiation dose. The rate, magnitude, and timing of suppression and accommodation were determined. The ability of periodic treatment with a lipopolysaccharide immunomodulator to alleviate the suppressive hematopoietic effects of chronic exposure was tested. The effects of other pharmacologics (amifostine, granulocyte colony-stimulating factor, cytokine) are projected on the basis of current research using rodent models. Results indicated that low but significant suppression of blood leukocyte and platelet levels occurred at 3 mGy per day. As the dose rate increased from 3 mGy to 128 mGy per day, the rate of suppression increased approximately eightfold, whereas the time to accommodate declined from 2,000 days to approximately 150 days. Within the time frame required to reach the upper limit of 700 mGy, none of the dose rates examined elicited blood cell decrements large enough to severely compromise near-term immune function. Pharmacological intervention with lipopolysaccharide minimized hematopoietic suppression in only a small fraction of the treated animals that displayed distinctive long-term survival and pathology patterns. Comparable short term benefits of treatment with hematopoietic cytokines or chemoprotectants are predicted on the basis of responses noted in rodent models. Long-term benefits of such treatments remain to be determined. Future work will require the application of advanced molecular tools to more fully assess potential pathophysiological responses and their modulation after low-level radiation exposure. PMID- 11873527 TI - Prevention and treatments: summary statement. PMID- 11873528 TI - Nuclear, biological, and chemical combined injuries and countermeasures on the battlefield. AB - The Armed Forces Radiobiological Research Institute (AFRRI) has developed a research program to determine the major health risks from exposure to ionizing radiation in combination with biological and chemical warfare agents and to assess the extent to which exposure to ionizing radiation compromises the effectiveness of protective drugs, vaccines, and other biological and chemical warfare prophylactic and treatment strategies. AFRRI's Defense Technology Objective MD22 supports the development of treatment modalities and studies to assess the mortality rates for combined injuries from exposure to ionizing radiation and Bacillus anthracis, and research to provide data for casualty prediction models that assess the health consequences of combined exposures. In conjunction with the Defense Threat Reduction Agency, our research data are contributing to the development of casualty prediction models that estimate mortality and incapacitation in an environment of radiation exposure plus other weapons of mass destruction. Specifically, the AFFRI research program assesses the effects of ionizing radiation exposure in combination with B. anthracis, Venezuelan equine encephalomyelitis virus, Shigella sonnei, nerve agents, and mustard as well as their associated treatments and vaccines. In addition, the long-term psychological effects of radiation combined with nuclear, biological, and chemical (NBC) injuries are being evaluated. We are also assessing the effectiveness of gamma photons and high-speed neutrons and electrons for neutralizing biological and chemical warfare agents. New protocols based on our NBC bioeffects experiments will enable U.S. armed forces to accomplish military operations in NBC environments while optimizing both survival and military performance. Preserving combatants' health in an NBC environment will improve warfighting operations and mission capabilities. PMID- 11873529 TI - Chemical toxicants and low-level radiation exposure. PMID- 11873530 TI - The incidence of injury in light infantry soldiers. AB - U.S. Army soldiers lose substantial training hours as a result of musculoskeletal injuries. Most military injury data have focused on basic trainee populations. This study examines activities associated with injuries in operational infantry soldiers. We conducted a randomized, retrospective medical record review of 339 records from a total of 3,195 light infantry soldiers over 13 months. The annualized incidence of injuries was 95 per 100 soldiers per year vs. 74 per 100 for illness. There were 372 injuries, representing 56% of sick-call diagnoses. Physical training caused 50% of all injuries, and 30% of those were linked to running. Injuries caused nearly 10 times the number of limited duty days as illness. Soldiers with lower extremity running injuries spent seven times more days on profile than those with nonrunning injuries. We conclude that physical training is related to a high number of injuries in infantry soldiers. This study provides important data on activities and injuries in operational infantry soldiers. PMID- 11873531 TI - Differences in length of stay between coronary bypass and valve procedures. AB - OBJECTIVE: Emphasis on cost reduction, national standardization of medical care, and quality improvement initiatives have led to reduced postoperative hospital stays after cardiac surgery. The present study was designed to verify the observation that valve patients have longer lengths of stay than bypass patients and to identify possible reasons. METHODS: The inpatient records of 26 consecutive patients who underwent valve procedures at our institution were reviewed and compared with the records of 25 consecutive coronary bypass patients. Patients whose postoperative stays were longer than 2 weeks were considered outliers and were excluded from further review. RESULTS: A total of 51 records were reviewed. There were no in-hospital deaths. Five patients in the valve group and two in the bypass group were excluded because of lengths of stay exceeding 14 days. The mean length of stay for the valve procedure group was 7.7 +/- 2.1 days, vs. 5.7 +/- 1.5 days for the coronary bypass group (p = 0.001). There were no reoperative procedures in either group, and the number of emergency procedures was higher in the bypass group. Both groups were similar with respect to age and sex. The frequencies of associated pulmonary disease and malnutrition were similar. Bypass patients had a higher incidence of vascular disease and/or renal disease. Sixty percent of valve procedures were complex operations. Valve patients had a higher incidence of cardiopulmonary bypass times exceeding 3 hours and more postoperative bleeding complications. Although not statistically significant, valve procedure patients were more likely to require prolonged mechanical ventilatory and circulatory support postoperatively. The frequencies of thrombotic complications, neurological complications, and nosocomial infections were similar for both groups. Postoperatively, more valve patients had atrial fibrillation, and all of them received anticoagulation. Multivariate analysis revealed only two factors to be significant with regard to length of stay: valvular surgery and the duration of postoperative ventilatory support. CONCLUSIONS: Cardiac valve procedures are associated with prolonged lengths of stay compared with isolated coronary bypass procedures. Despite recent trends toward earlier operation, valve patients tended to present with advanced disease. More than half of the valve patients required complex surgical procedures. This is reflected in longer bypass times, increased bleeding complications, and more postoperative support. The proportion of valvular surgery patients should be considered when analyzing lengths of stay for cardiac surgery cohorts. PMID- 11873532 TI - Testing a model of exposure to environmental tobacco smoke in military women with children. AB - The purpose of this study was to test a model of exposure to environmental tobacco smoke (ETS) in military women and their children. The convenience sample consisted of 238 women, 81 smokers and 157 nonsmokers, with a mean age of 37 years (SD = 9.9). Participants were either on active duty or were reservists and/or military dependents. Model constructs, some of which were adapted from the transtheoretical model of behavior change, measured personal and situational factors, pros and cons of ETS exposure, self-efficacy to resist ETS, mother's expectation for child's ETS exposure, and mother's self-efficacy to reduce child's ETS exposure. The mediating variable was the mother's daily ETS exposure, and the outcome variable was the child's daily ETS exposure. The trimmed model showed that 32% of the variance in mother's daily exposure (mediating variable) was accounted for by living with a smoker, having high ETS "pros" (as opposed to ETS "cons"), having less self-efficacy to resist ETS, and having greater self efficacy to reduce the child's exposure. There was a significant, positive relationship (r = 0.51, p = 0.01) between the mother's and child's daily ETS exposure (outcome variable). PMID- 11873533 TI - The Department of Defense's Persian Gulf War registry year 2000: an examination of veterans' health status. AB - This study examined the health status of 46,633 Persian Gulf War theater veterans who received full clinical evaluations in the Department of Defense's Gulf War Comprehensive Clinical Evaluation Program (CCEP) as of spring 2000. Clinical data analyzed included demographic information, 15 health symptoms, 19 wartime exposures, and primary and secondary physician-determined medical diagnoses based on International Classification of Diseases, 9th Revision, Clinical Modification, criteria. Findings and discussions are arrayed, by gender, with comparative 1996 data from the Department of Veterans Affairs Health Examination Registry Program. Many veterans reported fewer physical symptoms now than during the time of the Gulf War. Many endorsed symptoms of joint pain, fatigue, weight change, and sleep disturbances. Most reported exposure to diesel fuel and the nerve agent antidote pyridostigmine bromide; far fewer female veterans reported combat involvement. The most frequent primary or secondary diagnosed medical conditions were musculoskeletal/connective tissue diseases, ill-defined conditions, and mental disorders. Female veterans were diagnosed more frequently with mental disorders. Symptom endorsement and diagnosis rates between the CCEP and the Department of Veterans Affairs registry were not dissimilar. Overall, the self-reported general health of veterans with symptoms was much poorer (females had higher rates of "fair to poor" health than males) than that of veterans with no reported symptoms. PMID- 11873534 TI - Management of ectopic pregnancy at a military medical center. AB - OBJECTIVE: Advances in laparoscopic surgical techniques and ultrasound technology along with the popularization of treatment with methotrexate have revolutionized accepted algorithms for treating ectopic pregnancy. I analyzed the management of ectopic pregnancy at a military medical center. METHODS: A retrospective chart review of ectopic pregnancies at Womack Army Medical Center between January 1997 and July 1999 was conducted. RESULTS: A total of 129 cases of ectopic pregnancy were found. Of those, 77 patients received primary surgical management, 51 patients received primary therapy with methotrexate, and 1 patient was managed expectantly. Eleven patients failed methotrexate therapy and went on to surgical therapy. Of the 88 patients ultimately treated surgically, 76 underwent laparoscopic procedures, 12 received laparotomy, and 11 more converted from laparoscopy to laparotomy. CONCLUSION: The advent of methotrexate therapy has greatly facilitated the treatment of ectopic pregnancy and altered our institutional algorithm; however, primary surgery is still used more often. PMID- 11873535 TI - U.S. Navy and Marine Corps Hearing Conservation Program, 1995-1999: mean hearing thresholds for enlisted personnel by gender and age groups. AB - This study presents mean hearing thresholds from a cross-sectional study of 68,632 monitoring audiograms submitted to the Navy Environmental Health Center for 1995 to 1999. Records included U.S. Navy and U.S. Marine Corps enlisted personnel (Navy men = 51,643; Navy women = 4,184; Marine Corps men = 12,251: Marine Corps women = 554). Mean hearing thresholds were calculated for age groups (17-24, 25-29, 30-34, 35-39, 40-44, 45-49, and 50 years and older), gender (male/female), and service (Navy/Marine Corps). Although hearing thresholds worsened with increasing age, as expected, Navy and Marine Corps men have worse levels than Occupational Safety and Health Administration age-corrected values throughout most of their careers, whereas women were closer to the Occupational Safety and Health Administration values. Hearing thresholds in the Navy have not improved appreciably from historical reports. Threshold variability, as revealed by standard deviations, increased with age and paralleled threshold levels. This epidemiological information suggests that hearing conservation continues to be an important force health protection issue. PMID- 11873536 TI - Acute pulmonary effects from o-chlorobenzylidenemalonitrile "tear gas": a unique exposure outcome unmasked by strenuous exercise after a military training event. AB - o-Chlorobenzylidenemalonitrile, more commonly called CS, is grouped with several other irritant agents referred to as "tear gas." It is a riot-control agent used frequently in military settings to test the ability and speed of personnel in donning their military gas masks. When personnel are exposed to CS without proper personal protective equipment, it has potent irritant effects. We report a unique cluster of hospitalizations of nine U.S. Marines who developed a transient pulmonary syndrome. All nine patients had symptoms of cough and shortness of breath. Five of the nine presented with hemoptysis, and four presented with hypoxia. Symptoms were associated with strenuous physical exercise from 36 to 84 hours after heavy exposure of CS in a field training setting. Four of the nine Marines required intensive care observation as a result of profound hypoxia. All signs and symptoms resolved within 72 hours of hospital admission. One week after CS exposure, all nine Marines demonstrated normal lung function during spirometry before and after exercise challenge using cycle ergometry. PMID- 11873537 TI - Predictors of course completion for the U.S. Army pharmacy specialist course. AB - The purpose of this study was to determine predictors of successful completion of the U.S. Army Medical Department pharmacy specialist (91Q) training program. The sample consisted of 143 students from four course iterations, and only students enrolled in the course for the first time were included for analysis. The relationship between 16 predictor variables and successful completion was assessed. Five variables, the ranks of private (E1) and specialist (E4), cross trainee status, gender, and Armed Services Vocational Aptitude Battery (ASVAB) general technical (GT) score, were found to be significant predictors using simple linear regression. Using hierarchical multiple linear regression analysis, three variables, service component, gender, and ASVAB GT score, were found to have a significant unique shared variance with success. The results of this study indicate that ASVAB GT scores are predictive of success (p < 0.01) in the 91Q training program. Using these predictors, the selection process may be modified to improve the likelihood of student success in the course. PMID- 11873538 TI - A behavioral intervention to prevent sexually transmitted diseases/human immunodeficiency virus in a Marine Corps sample. AB - Research evaluating human immunodeficiency virus (HIV) behavioral interventions among U.S. military personnel has been lacking. In this study, a behavioral intervention to prevent HIV and sexually transmitted diseases (STDs) was implemented in a sample of Marine security guards. Participants were assessed before and after a three-session intervention on a measure of STDs/HIV knowledge and a number of psychosocial scales. The results indicated that STDs/HIV knowledge was significantly greater after the intervention. Significant pre-test vs. post-test differences were also found on the Social Norms, Behavioral Intentions, Attitudes toward Condoms, and Self-Efficacy/Impulse Control scales. On Social Norms and Behavioral Intentions, the differences were as expected: subjects perceived greater social norms supporting condom use and had stronger intentions to practice safe sex after the intervention than they had before. On Attitudes toward Condoms and Self-Efficacy/Impulse Control, the differences were not in the expected direction. Psychosocial factors associated with self-reported condom use were also identified. PMID- 11873539 TI - Ultrasound, a new tool for surface matching in computer-navigated surgery. AB - OBJECTIVE: The object of this study was to investigate the feasibility of generating a bone surface from data provided by an ultrasound examination and to match this surface with the previous computed tomography (CT) scan. METHODS: From a CT data set of a training model of the pelvis, a three-dimensional surface was extracted by global thresholding-based segmentation. The same model was placed in a water basin, and ultrasound images were taken with a guided ultrasound transducer. The three-dimensional surface was generated from the ultrasound data set, and the two surfaces were matched in a semiautomatic mode. RESULTS: With special segmentation methods, a surface could be extracted automatically from the CT and the ultrasound data set. From these segmented ultrasound slices, a volume data set of the model was generated. After approximate initial matching, the local matching process was completed automatically. CONCLUSION: One of the limitations in computer-assisted surgery is the complicated matching process. Using special algorithms, a surface was extracted from the data set of an ultrasound examination and matched in a semiautomatic mode with the surface of a CT data set, facilitating the matching process. PMID- 11873540 TI - "Ecstasy"-induced hepatitis in an active duty soldier. AB - 3,4-Methylenedioxymethamphetamine, or "Ecstasy," is a drug commonly used at "rave" parties to heighten energy and intimacy. Although its complications have been well described in Europe, including less common side effects such as hepatic failure and rhabdomyolysis, physicians in the United States have less experience with this drug because of the shorter duration of its use in this country. We present a case of an active duty soldier who was admitted for acute hepatitis secondary to Ecstasy ingestion and describe its proposed pathophysiology. We believe that with the increased use of Ecstasy in the United States, especially among younger patients, including soldiers, military physicians will need to be more familiar with this potentially deadly drug. PMID- 11873541 TI - Magnetic resonance imaging findings in a case of cytarabine-induced myelopathy. AB - A 44-year-old white male with an isolated central nervous system relapse of acute lymphoblastic leukemia was treated with intrathecal cytarabine. He developed Staphylococcus epidermidis meningitis, which was treated successfully with intrathecal vancomycin. Four weeks after the initiation of intrathecal cytarabine, the patient developed progressive ascending paralysis to the upper cervical level. Initial magnetic resonance imaging of the brain and spine were normal, and cerebrospinal fluid evaluation showed no evidence of ongoing infection and clearance of lymphoblasts. Three weeks later, magnetic resonance imaging demonstrated marked edema and peripheral enhancement of the spinal cord, consistent with cytarabine toxicity. PMID- 11873542 TI - Oropharyngeal hymenoptera stings: a special concern for airway obstruction. AB - Hymenoptera stings are common and cause 40 to 50 deaths each year. Hymenoptera venom contains a variety of toxic and allergenic substances that can produce many types of both local and systemic reactions. Of these, anaphylaxis is the most feared and the most common cause of sting-related deaths. Oropharyngeal stings, although rare, have the added potential to produce life-threatening airway obstruction via localized swelling. This threat is of particular concern to military personnel who operate in environments where stings are more likely to occur and where emergency medical resources are limited or lacking. This risk can be minimized if such victims are treated early and aggressively, even though they may initially present with minimal symptoms. PMID- 11873543 TI - Acute presentation of undiagnosed end-stage renal disease in a young active duty male soldier. AB - Chronic renal insufficiency is a condition involving the deterioration of renal function over a period of months to years. This differs from acute renal failure, which is a sudden decrease in renal function that occurs over a period of hours to days. Patients with chronic renal insufficiency are often clinically asymptomatic until nearing end-stage renal disease, at which time their signs and symptoms reflect subtle changes in metabolic and volume control. In contrast, acute renal failure patients often present with overt symptoms caused by sudden metabolic abnormalities and volume overload. We describe a patient who's clinical presentation suggested acute renal failure but was actually caused by end-stage renal disease from a chronic renal process. PMID- 11873544 TI - Deep partial thickness burn after contact with a Meal Ready-To-Eat heater. AB - A case of a Meal Ready-To-Eat heater injury resulting in a deep partial thickness burn is presented. The patient was treated as an outpatient at our battalion aid station. By establishing the depth, extent, and severity of a burn injury, a decision can be made regarding whether outpatient or inpatient therapy is appropriate. A brief description of the Meal Ready-To-Eat heater injury and general guidelines for the outpatient management of burns are presented. PMID- 11873545 TI - Takayasu arteritis presenting as a recurrent respiratory tract infection: a diagnosis facilitated by bedside echocardiography and increased erythrocyte sedimentation rate. AB - Takayasu arteritis is a rare autoimmune disease affecting large and moderate sized arteries, often involving the aorta or coronary vasculature. We report a case of an adolescent male with a history of recurrent respiratory tract infections who presented with fever, cough, and shortness of breath and who was diagnosed with acute aortic valve failure and coronary ischemia. Ultimately, the patient's condition was attributed to Takayasa arteritis. This typical presentation of an atypical disease provides valuable teaching points, including the use of bedside echocardiography for the diagnosis of acute aortic insufficiency and the differential diagnosis of increased erythrocyte sedimentation rate. It also serves to remind clinicians to maintain a high index of suspicion for unusual disease processes in patients who fail to respond to empiric therapy for recurrent subacute illnesses. PMID- 11873546 TI - Coxa saltans externa treated with Z-plasty of the iliotibial tract in a military population. AB - Coxa saltans, or snapping hip, can be attributable to a number of causes. Coxa saltans externa typically occurs when the thickened portion of the iliotibial band snaps over the greater trochanter as the hip is flexed. This condition generally resolves with a course of nonoperative treatment. We have treated three patients with Z-plasty of the iliotibial band, which has the highest published success rate in the English language literature. Only one of three patients was able to return to full activities postoperatively. In our small series, this method of surgical treatment in an active duty military population yielded less than optimal results. PMID- 11873547 TI - A 20-year-old male with fever and hearing loss. PMID- 11873548 TI - Could foot and mouth disease be a biological warfare incident? PMID- 11873549 TI - Guidelines for evaluation of patients at risk for inherited breast and ovarian cancer: recommendations of the Department of Defense Familial Breast/Ovarian Cancer Research Project. AB - Patients at high risk for inherited breast and/or ovarian cancer are frequently encountered in all medical specialties. Department of Defense, Health Affairs funding as part of the Breast Cancer Education and Awareness Program was used to develop a comprehensive program for the identification, counseling, genetic testing, and long-term follow-up of such high-risk patients. This article reports the recommendations for high-risk patient management based on 4 years of evaluation and care, including discussions of the approach to counseling, indications for genetic testing, post-testing counseling, patient surveillance with examination, imagining, and laboratory testing, and suggested options for surgical and chemoprophylaxis as well as lifestyle modifications. PMID- 11873550 TI - Diagnostic criteria for testing for BRCA1 and BRCA2: the experience of the Department of Defense Familial Breast/Ovarian Cancer Research Project. AB - The Department of Defense Familial Breast/Ovarian Cancer Research Project has offered genetic counseling and testing for BRCA1 and BRCA2 on a research basis to patients meeting specific diagnostic criteria, with risk for BRCA1 and BRCA2 mutations calculated based on the Couch model. In 2.5 years, 250 patients were evaluated and 101 patients met criteria requirements, including 33 who met criteria in more than one category. Ninety patients elected to undergo DNA testing. In this group of 90 patients, 14 mutations (15.5%) and 16 unclassified variants (17.7%) were identified. The most common inclusion criteria were onset of breast/ovarian cancer before age 45 years (n = 32) and onset of breast/ovarian cancer before age 45 years with strong family history (n = 21). However, when number of mutations and unclassified variants found were compared separately across all diagnostic criteria (including those of more than one capacity) using the chi 2 statistic, no significant differences were seen among the categories to suggest that one criterion was more predictive of mutations or variants than another. Couch risk values for patients with mutations showed a mean of 14% and ranged from 3.2 to 43.5% (range for all patients, 1.2-69.7%). These findings emphasize the importance of using multiple diagnostic criteria and suggest that a Couch risk value of > 3% may be useful in selecting patients for testing. The data also underscore the necessity of genetic counseling in the testing process, particularly given the large number of unclassified variants diagnosed and their uncertain status for disease predisposition. PMID- 11873551 TI - AIDS fund board forms, issues first guidelines for awarding grant funds. PMID- 11873552 TI - Bush favors boosting AIDS research, but other funding static. PMID- 11873553 TI - World economic forum urges more support for AIDS fight. PMID- 11873554 TI - WHO: world may be mobilizing for battle against HIV/AIDS. PMID- 11873555 TI - New program seeks sponsors for country-based projects. PMID- 11873556 TI - Bush's new pledge to global AIDS fund said too small. PMID- 11873558 TI - Convicted murderer can avoid determination of HIV status. PMID- 11873557 TI - Planned namming of AIDS council cochairman stirs criticism. PMID- 11873559 TI - Women face higher HIV/AIDS risks, lower treatment rates. PMID- 11873561 TI - Appeals court decision sends viatical-fraud case to Florida. PMID- 11873560 TI - Court upholds penalty for refusal to take blood-alcohol test. PMID- 11873562 TI - Judge allows housecleaning needlestick lawsuit to proceed. PMID- 11873563 TI - HIV-positive woman loses right to visit her granddaughter. PMID- 11873564 TI - Court: woman failed to file timely suit after blood exposure. PMID- 11873565 TI - [Good intentions--hard realities]. PMID- 11873566 TI - [Tobacco kills]. PMID- 11873567 TI - [Can diabetes be prevented?]. PMID- 11873568 TI - [Physicians, sick leave and occupational life included]. PMID- 11873569 TI - [Does systematic evaluation of sickness certification II lead to less use of health insurance?]. AB - BACKGROUND: The increasing prevalence of disability pensioning in Norway has led to several attempts at strengthening the proactive role of the National Insurance System (NIS) in cases of long-term sick-listing. Since 1988, a special medical certificate is required after eight weeks of sick-leave. The aim of this study was to examine whether systematic evaluation of this medical certificate by NIS officers and NIS medical consultants could reduce future health insurance expenditure. MATERIAL AND METHODS: In 1994 a randomised study using a paired design of the NIS local offices in the county of Hordaland was undertaken. All eight-week medical certificates in the intervention group (N = 2,237) were systematically reviewed, whereas standard routines were used for the control group (N = 1,764). RESULTS: After three years, no significant differences were observed between the two groups in health insurance utilisation. INTERPRETATION: We conclude that local NIS offices are unable to use the information in the eight week sick notes to effectively influence future utilisation of health insurance. The reason may be that NIS offices lack the skills necessary for early intervention in long-term sick-listing. PMID- 11873570 TI - [Assessing the quality of medical certificates II--useful criteria or unnecessary schedule?]. AB - BACKGROUND: In Norway, doctors must provide the National Health Insurance (NIS) with a special medical certificate if a sick-leave exceeds eight weeks. The aim of this study was to evaluate this medical information in relation to the NIS criteria for further sickness benefits and the usefulness of the certificate in assessing the need for early rehabilitative and vocational initiatives. MATERIAL AND METHODS: 2,237 consecutive eight-week medical certificates submitted to 21 NIS offices in the county of Hordaland in 1994 were independently evaluated by the NIS officer and the NIS medical consultant in a questionnaire survey. RESULTS: Both evaluators found sufficient medical information in the vast majority of medical certificates, though in about 12% of the cases the medical criteria for further benefits were not sufficiently documented. In less than 10% of the cases, active rehabilitative or vocational measures were called for at this point in time. However, there was a rather low degree of consensus between the NIS officer and the medical consultant about in which cases supplementary information or active measures were warranted. INTERPRETATION: We conclude that the eight-week medical certificate usually contains enough medical information but is of limited use in assessing the need for active intervention by the NIS. Supplementary information from the absentee and the employer should be collected at an early stage of the sick leave. PMID- 11873571 TI - [Low-grade non-Hodgkin lymphoma in Northern Norway]. AB - BACKGROUND: We present a study of treatment outcome and evaluate pre-treatment prognostic factors in patients treated for low-grade non-Hodgkin's lymphoma (LG NHL) at our institution during a ten-year period. MATERIAL AND METHODS: 169 consecutively registered patients from 1986 to 1996 were retrospectively analysed with regard to demographic, treatment, and disease-specific characteristics. Median follow-up time was 52 months. All patients were diagnosed histologically according to the previous Kiel classification system. Median age was 60 years; the male:female ratio was 1.05:1. RESULTS: The response rate was 66%; median overall survival 8.3 years. Five and ten-year overall survival were 72% and 47% respectively. For stage 1, 2, 3 and 4, ten-year overall survival were 86%, 65%, 33% and 29%. Of the pre-treatment parameters, advanced stage, B-symptoms, poor performance status, bone marrow infiltration, tumour > or = 6 cm, low serum albumin, anaemia, and LD > or = 540 U/l were all related to survival. According to the multivariate analysis, stage, performance status, tumour size, and anaemia were found to be independent prognostic factors for overall survival. INTERPRETATION: The strategy of radiation therapy has proved successful for most patients with stage 1 disease. Independent prognostic factors for overall survival such as stage, performance status, tumour size, and anaemia may be useful guides in deciding when and how to treat. PMID- 11873572 TI - [A patient with thromboangiitis obliterans]. AB - BACKGROUND: Thromboangiitis obliterans (Buerger's disease) is characterised by inflammatory thrombi in small and middle-sized arteries and in the veins. As opposed to arteriosclerosis, the vessel wall remains normal. Diagnostic criteria are the presence of distal ischaemia in the extremities, documented by noninvasive vascular testing; exclusion of autoimmune diseases, hypercoagulable states, and diabetes mellitus, age less than 45 years and cigarette smoking. MATERIAL AND METHODS: The article is based on a case history and review of the literature. RESULTS AND INTERPRETATION: A 31-year-old man was in 1953 diagnosed with thromboangiitis obliterans. Lumbar sympathectomy on the left side in 1953 and on the right side in 1971 had excellent symptomatic effect. The patient has been treated with anticoagulants for 48 years. Amputation of the left 2nd toe was performed in 1971. At age 80, he is without further cardiovascular events and takes regular physical exercise. Over the last fifty years, the incidence of thromboangiitis obliterans has been drastically reduced. The reason for this is unknown. The only proven treatment is complete discontinuation of tobacco. The effect of sympathectomy and long-term anticoagulation has not been proven. PMID- 11873573 TI - [Aerococcus urinae and urinary tract infection]. AB - BACKGROUND: Bacteriological investigation of urine often shows the presence of alpha-haemolytic streptococci, but the clinical significance of this finding may be unclear. Among the alpha-haemolytic streptococci found in urine is Aerococcus urinae, which is pathogenic. MATERIAL AND METHODS: We present a prospective study of the prevalence and antibiotic susceptibility of A urinae in urine samples sent to a microbiology laboratory in Norway. RESULTS: Seven out of 27 alpha-haemolytic streptococcal isolates were A urinae. This represents 0.3% of all urine isolates from our laboratory. Appropriate methods for identification and resistance determination are described. INTERPRETATION: Microbiological laboratories should include identification of A urinae in their diagnostic repertoire. Recommended treatment against A urinae is ampicillin or nitrofurantoin. PMID- 11873575 TI - [Acute cerebral infarction--diagnosis of subgroups]. AB - BACKGROUND: Subclassification of cerebral infarcts is crucial in order to optimise and differentiate therapy. If we are to apply specific treatment modalities early while neurological deficits are still reversible, we need a fast diagnosis that reliably predicts etiology. MATERIAL AND METHODS: The early clinical classification into Oxford subgroups is presented with an assessment of their predictive value in relation to aetiology (TOAST classification). RESULTS: The various Oxford subgroups correlate well to the topography of the infarction demonstrated by neuroradiology, to the vascular aetiology as demonstrated by ultrasound techniques, and to the "aetiologic" subgroup as determined by a full diagnostic work-up. However, the correlation is not sufficient to guide potentially specific treatments of subgroups if the treatment in question has serious side effects (e.g. thrombolysis). However, if the clinical diagnosis is supplemented with diffusion-weighted MRI and perhaps MR angiography, the predictive value of the diagnosis is extremely high. INTERPRETATION: Very early clinical diagnosis of cerebral infarction must be supplemented with diffusion weighted MRI to make the diagnostic precision sufficiently high to guide the application of specific treatments with potentially serious side effects. PMID- 11873574 TI - [Tyrosine kinase receptor-ras-ERK signal transduction pathway as therapeutic tarfet in cancer]. AB - BACKGROUND: Experimental evidence indicates that various intracellular signalling cascades are altered in tumour cells. Among these, the receptor tyrosine kinase ras-ERK signalling pathway was found to be constitutively active in a significant percentage of human tumours; hence, considerable effort has been directed at finding compounds that inhibit its activation. MATERIAL AND METHODS: We review the recent progress in establishing novel approaches to interference with the constitutive activation of the receptor tyrosine kinase ras-ERK signalling pathway in cancer. RESULTS: Inhibition of the receptor tyrosine kinase ras-ERK signalling pathway activation by various novel agents (e.g. small molecule tyrosine kinase inhibitors, antibodies, FTase inhibitors, SH2/SH3 directed agents, antisense, ribozymes) impaired tumour growth. Some of the developed agents have been tested in clinical trials; promising results were obtained. INTERPRETATION: Inactivation of the receptor tyrosine kinase ras-ERK signalling pathway by small molecular inhibitors has confirmed its involvement in tumour growth. Thus, molecular and/or pharmacological modulation of the components that are critically involved in the constitutive activation of this pathway are expected to improve the treatment of human malignancies. PMID- 11873576 TI - [Pharmaceutical revision of hospital drug administration]. AB - BACKGROUND: Quality audits of the implementation of drug administration procedures are carried out in order to determine objectively to what extent implementation conforms to procedures. METHODS: Since September 1997, the pharmacy at Rikshospitalet University Hospital in Norway has performed quality audits of drug administration at the hospital, using interviews and surveys. Staff members in the audited unit and the auditing pharmacist agree on prospects for quality improvements and review possible action. A survey was carried out in the autumn of 2000 in order to determine staff opinion of the quality audits. RESULTS: On the basis of the observations made, improvements have been carried out at all levels of the organisation. INTERPRETATION: The survey indicates that hospital staff members are satisfied with the quality audits performed by the pharmacy. PMID- 11873577 TI - [Anticoagulation therapy of atrial fibrillation]. PMID- 11873578 TI - [Smoking in Norwegian hospitals--status by the end of the year 2000]. AB - BACKGROUND: Establishing smoke-free hospitals by the end of 1995 was a goal for Norwegian health authorities. The present study reports the smoking status in Norwegian hospitals by the end of the year 2000. MATERIAL AND METHODS: Information was collected by questionnaires sent to all 81 Norwegian hospital directors; 78 (96%) responded. RESULTS: 19 hospitals (27%) were still not smoke free. The lowest smoke-free rate (60%) was reported from psychiatric institutions. Smoking in all office areas was prohibited while 10% of all hospitals allowed smoking in their cafeterias. Outdoor smoking areas were provided in two thirds of the hospitals. Separate smoking rooms for patients were more common in non-smoke-free hospitals (79%) than in smoke-free institutions. The smoking restrictions were accepted with little resistance, but illicit smoking was not uncommon, even in-house. Sale of tobacco and cigarettes was a widespread practice in the hospitals (63%), irrespective of their smoking policy. INTERPRETATION: It appears that there is still a long way to go before Norwegian hospitals are really smoke-free. Sale of tobacco and cigarettes should be banned and further restrictive measures from the government are required together with active participation from doctors and nurses as role models and not the least from the patients themselves, in their own best interest. PMID- 11873579 TI - [Smoking ban in psychiatric department]. AB - BACKGROUND: It is well known that nicotine and caffeine can increase tension and anxiety, disturb sleep and cause a variety of smoking related health problems. MATERIAL AND METHODS: On 1 March 2001, it was decided to ban smoking in a general psychiatric inpatient unit and the patients were asked to confine smoking to outdoor areas. In a retrospective study we looked at the effects of the ban on smoking on "as needed" medication, ward atmosphere and incidents related to the change. The records from 30 patients discharged before the ban and 30 patients after the ban were examined and the demands for "as needed" medication registered. RESULTS: Two-sample t-test analyses of the groups showed a significant difference in "as needed" medications used between the two groups, with a reduction in the group discharged after the smoking ban (p < 0.01). There was a substantially reduction in coffee consumption and positive improvement in ward atmosphere after the ban as observed by the staff. INTERPRETATION: The material is limited; others are encouraged to proceed with validation of our experience. PMID- 11873580 TI - [Air pollution and health]. PMID- 11873582 TI - [Infant memory difficult to catch]. PMID- 11873581 TI - [Sexual abuse, recovered memory and multiple personality disorder]. PMID- 11873583 TI - [Does organized mammographic screening reduce breast cancer mortality? The Cancer Registry's view on the current debate]. PMID- 11873584 TI - [Therapeutic cloning--a breakthrough without scientific evidence?]. PMID- 11873585 TI - [Chronic pain--everything we do is not wrong]. PMID- 11873586 TI - [Derailment concerning generic substitution]. PMID- 11873587 TI - [Rehabilitation of psychiatric long-term patients]. PMID- 11873588 TI - [Alf Brodal--a great teacher]. PMID- 11873589 TI - [Medical effects of transcendental meditation]. PMID- 11873590 TI - [Arvid Carlsson: a Nobel Prize winner in physiology or medicine in the year 2000]. PMID- 11873591 TI - [Neurological and psychiatric symptoms in patients with systemic lupus erythematosus]. AB - Sixty patients (51 women/9 men) with diagnosis of SLE were studied for finding out the frequency of nervous system involvement in SLE, the time of of appearance of neurological involvement after diagnosis establishing, the coexistence of the antiphospholipid syndrome, and the character of changes in MR, CT and CSF. Nervous system involvement was found in 40 cases (67%), with 34 cases (56%) had involvement of the CNS, 6 patients (10%) had symptoms of peripheral nervous system dysfunction, and 3 (5%) had involvement of both systems. In 4 cases polineuropathy and transverse spinal cord lesion, and in 3 cases psychiatric symptoms were the first manifestations of SLE. Changes due to involvement of cerebral vessels (TIA, stroke) were observed in 20 patients (33%), and psychiatric symptoms in 16 cases (26.6%). No difference was found in the occurrence of stroke or TIA between SLE patients with and without antiphospholipid syndrome, and no correlations were noted between the presence of neurological or psychiatric symptoms and other SLE symptoms. CT scans demonstrated corticosubcortical atrophy in 28.3% of cases, while in MRI in T2 images small hyperintense lesions were situated mainly in the white matter (33.9%). In 5 cases oligoclonal band was found in the CSF, but without any correlation with specific neurological symptoms. PMID- 11873592 TI - [Thrombogenesis in patients with ischemic stroke]. AB - The activity of thrombinogenesis process is shown by thrombin restrain reaction by its physiological inhibitor antithrombin III, as a result of which biologically nonactive thrombinantithrombin III complexes (TAT) are created. The aim of the study was to evaluate TAT complexes in the blood of ischaemic stroke patients. 25 persons with ischaemic stroke and 15 controls took part in the examination. Taking into consideration brain CT outcomes 2 groups of patients were selected: with ischaemic stroke of small and great extent. The patients were also divided into a group of patients with less severe and with more severe ischaemic stroke, and into a group of patients with ischaemic stroke with and without accompanying atrial fibrillation. TAT complexes were determined with an enzyme-linked immunosorbent assay method on days 1st, 5th, and 12th after ischaemic stroke onset. TAT complexes were significantly higher in patients with cerebral infarction than in controls, significantly higher in patients with ischaemic stroke of great extent compared to patients with ischaemic stroke of small extent, and significantly higher in patients with more severe cerebral infarction than in patients with less severe cerebral infarction. In patients with cerebral infarction and atrial fibrillation TAT complexes were significantly higher only on the 1st day of the disease compared to patients with cerebral infarction without atrial fibrillation. Our investigations confirm intense activity of thrombinogenesis process in ischaemic stroke patients. PMID- 11873593 TI - [Neuroborreliosis: CT and MRI findings in 14 cases. Preliminary communication]. AB - Since 1987 when Januszkiewicz and Kieda first described borreliosis, it is commonly recognized as infection of the nervous system in Poland, especially in north-east region. The diagnosis of the disease is mainly based on typical clinical signs, supported by serological testing. In 14 patients with clinical symptoms of neuroborreliosis CT and MR were performed to evaluate CNS changes. MR examinations were abnormal in 36%. Most patients (60%) presented cerebral atrophy. In 2 cases areas of abnormal signal were identified within cerebral white matter as well as within the brain stem. In the first case it was, recognized as demyelination focus, in second one MR showed evidence of or were suggestive of vascular involvement. In one case symmetrical calcifications were also found in internal capsules. Neuroradiological signs in Lyme disease are not specific. Neuroborreliosis has to be considered when patients present foci of hyperintense signal (T2-weighted images) in white matter and brain stem. PMID- 11873594 TI - [Interaction evaluation between APOE epsilon 4 alleles and hippocampal atrophy. Preliminary communication]. AB - In CT images the atrophy of medial parts of temporal lobes was evaluated in 60 patients with Alzheimer-type dementia. In the genotype of 30 patients at least one APOE epsilon 4 allele was identified, the other 30 patients served as the control group. The results of in vivo measurements of temporal lobe atrophy in the examined patients did not show statistically significant relations with APOE genotype which was probably due to entirely different nature and application of both tests. PMID- 11873595 TI - [Sleep disorders as a risk factors for stroke]. AB - Sleep-disordered breathing seems to play a role in pathogenesis of stroke: for example, it was showed that snoring is a risk factor for stroke. Sleep disorders and sleep-disordered breathing increase the risk for stroke, probably by influencing systemic and cerebral blood circulation, causing hypoxaemia during night. This theory is supported by the fact of higher prevalence of stroke in the morning. During REM sleep there is a higher requirement for oxygen; as most sleep apnoeas occur during REM, it is possible that there is a relative hypoxaemia during this sleep stage. Stroke, including hemispheric stroke, can cause or aggravate the pre-existing sleep-disordered breathing. There are contradictory data in the literature regarding the influence of stroke on sleep architecture. Sleep disorders are associated with poorer stroke outcome, so their detection and treatment can be important in secondary stroke prophylaxis and will improve the patient's functioning and quality of life. PMID- 11873596 TI - [Corpus callosum pathological lesions in computed tomography and magnetic resonance imaging]. AB - The spectrum of congenital anomalies and other pathologic changes of the corpus callosum is presented. Callosal agenesis and hypoplasia, lesions in cases of congenital neurometabolic disorders, traumatic, vascular, inflammatory and neoplastic conditions as well as callosal atrophy are discussed. CT and MR images from the authors' own experience illustrate the material. PMID- 11873597 TI - [New data concerning the significance of immunoglobulins and B cells in the pathogenesis of multiple sclerosis]. AB - Contemporary studies concerning the significance of immunoglobulins in the mechanism of multiple sclerosis are based on the analysis on the genome level. It was found that genes encoding immunoglobulins are restricted to active gene segments and showed a great frequency of mutations in sequences encoding the complementarity-determining regions (CDR s). The above presented results led to the conclusion, that synthesis of immunoglobulins and activation of B cells are not only the result of nonspecific reaction but are the antigen-driven response, related to the immunological mechanism of the disease. PMID- 11873598 TI - [Multiple sclerosis in the year 2000: advances in the treatment with interferon beta-1b]. AB - The decline of millennium brings forward new challenges in therapeutic immunomodulation of relapsing-remittent multiple sclerosis (RR MS). Interferon beta-1b (Betaferon) belongs to expanding family of soluble cytokines which are capable to modify beneficially cellular immunity in RR MS. IFN beta-1b is indicated for young or middle-aged, ambulant patients with RR MS who have frequent relapses, "aggressive" brain lesions in magnetic resonance imaging and higher levels of pro-inflammatory cytokines. Betaferon reduced in people with RR MS mean annual rate and severity of relapses (34% and 49%) and lengthened median time to second relapse (to 295 days). IFN beta-1b also increased the number of relapse-free patients (31%). Although the drug diminishes weakly progression and activity (disability) it might improve slightly participation (handicap) and quality of life. The course of treatment, withdrawals from the trials, side effects and the occurrence of neutralizing antibodies to IFN beta-1b are concisely described. Combination of IFN beta-1b with other immunotherapeutic drugs needs further clinical trials. PMID- 11873599 TI - [Spontaneous spinal epidural hematoma with uncommon clinical presentation: a case report]. AB - Spinal epidural haematoma (SEH)--posttraumatic or spontaneous (SSEH) is a rare entity, usually associated with severe neurological deficit. Urgent surgical decompression of the spinal cord is mandatory procedure. A case of SSEH with unusual transient clinical symptomatology and subsequent spontaneous resolution during conservative management is reported. PMID- 11873601 TI - [Brain stereotactic procedures in the Department of Neurotraumatology CM UJ in Cracow]. AB - Stereotactic procedures using Leksell frame have been performed in that Department since 1995, and as jet 199 such operations were carried out. The most frequent complication after stereotaxic biopsies was intracerebral haemorrhage occurring in 0.6 to 7.2% of cases, according to various authors. In the presented material it occurred in 1.3% of biopsies. Besides biopsy, the stereotactic procedures (105 cases) were applied for reducing the size or removal of tumours, including multiple tumours in one or both hemispheres, in one-step operations. The histological diagnoses were based on the examination of HE-stained smears, immunocytochemical reactions and electron microscopic studies. The effectiveness of the operations was assessed in control examinations (CT). The greatest group of cases undergoing stereotaxic surgery were metastatic tumours (47 cases, 61.3%), primary brain tumours were treated in 22 cases (28.57%), meningiomas in 4 (5.19%), abscesses in 3 cases (3.89%). In patients with metastatic tumours improvement was obtained in 97% of cases. One patient died from disseminated malignancies with cardiorespiratory failure. In the remaining patients no worsening of the objective status was noted after stereotaxic surgery. PMID- 11873600 TI - [Evaluation of results of surgical treatment of epidural canal lipomatosis with 3 years of follow-up of a case]. AB - We present the results of surgical treatment of idiopathic epidural lipomatosis in a 38 year-old man. Our results were evaluated 3 years; after operation. This is the third case of idiopathic epidural lipomatosis in literature in which a very good result of treatment was obtained and patient returned to previous work. PMID- 11873602 TI - [Technical and diagnostic difficulties in stereotactic biopsy in children]. AB - The paper presents technical and diagnostic problems associated with stereotactic biopsies in children. Between the years 1992 and 2000, in the Dept. of Neurosurgery, Child's Health Center, we performed 132 stereotactic procedures, including 110 biopsies. In 15 cases bleeding developed from the biopsy site; in this group we performed 3 craniotomies for open haematoma evacuation, 1 child died (mesencephalic glioblastoma multiforme). The remaining 2 children recovered with no additional neurologic sequelae related to this complication. Since 1997 we use the Nashold needle to obtain tissue samples. This enables us to take larger and fewer samples with less risk of bleeding. The biopsy was non diagnostic in 3 cases. In 1 case we observed a slight movement of the child's head in relation to the stereotactic frame when the patient was moved from the CT scanner to the operating theater. In order to avoid such problems in the future and to facilitate transposition of heavy and paretic or unconscious patients, we devised a hard board with side handles. Methods designed to overcome these and other difficulties are described. PMID- 11873603 TI - [Stereotactic biopsy: safety based on own experience]. PMID- 11873604 TI - [Pathomorphological evaluation in stereotactic brain tumor biopsy]. AB - The method is described of multistep pathomorphological examination of biopsy material obtained by stereotactic procedures from brain tumours. In the analysis of 260 biopsies the possibility was assessed of intraoperative cytological examinations confronting the results of these examinations with those of final histological and immunohistochemical investigations. Attention is called to the difficulties in the evaluation of material derived from stereotactic biopsies and to the cause of possible diagnostic errors. It is concluded that the combination of the results of cytological examination of smears and histological examination of paraffin slides makes possible establishing of full pathomorphological diagnosis with determination of tumour type and its malignancy degree in 90.7% of cases. On most cases cytological intraoperative examination had the greatest influence on therapeutic management. In the confrontation of these examinations with histological examinations a high proportion of correct cytological examinations was found (76.6%). Owing to the possibility of application of all techniques of morphological diagnosis, stereotactic biopsy is a reliable method in the diagnosis of brain tumours. PMID- 11873605 TI - [Diagnostic pitfalls in stereotactic biopsy]. AB - Between October 1998-August 2000, 65 patients were diagnosed, and underwent stereotactic biopsy of brain lesions. Procedures were performed with the Riechert Mundinger stereotactic device. The diagnosis was made intra-operatively, by examination of cytological smears. The initial diagnosis was confirmed after H&E, and selective stains, as well as immunohistochemical examinations of paraffin specimens. The method applied makes possible to obtain a reliable, and precise morphological diagnosis in the majority of cases, so that a proper treatment method could be introduced. PMID- 11873606 TI - [Stereotactic biopsy of polylmorphic tumors and brain tumors invisible in CT image]. AB - The authors present the possibilities of obtaining tissue samples with stereotactic biopsy basing on three examples of intracranial polymorphic and invisible in CT tumours. CT scans of these tumours make a serious problem with proper targeting and tissue sampling from proper sites. Stereotactic serial biopsy directed by CT/MRI fusion is the best way of obtaining tissue material in order to establish correct diagnosis. PMID- 11873607 TI - [Considerations of biopsy in neurological diagnosis]. AB - The evolution is described in the views of neurologists on the indications to brain biopsy in clinical practice. In the 1960 and 1970s the main indication were diffuse progressing processes in brain associated with dementia. Presently, this biopsy is done mainly in cases of focal lesions of unclear aetiology. Technical advances in neurovisual examinations and the development of stereotactic methods sparing the patients open presently extensive possibilities of neuropathological diagnosis of lesions, even those situated deeply in the brain. The ethical and legal principles of the use of biopsies in neurological practice have not changed over years. PMID- 11873608 TI - [Early immunomodulatory treatment of multiple sclerosis. Economic aspects]. PMID- 11873609 TI - [Report on the 14th Congress of the European Society of Functional and Stereotactic neurosurgery --ESSFN, London, October 25-27, 2000]. PMID- 11873610 TI - [Physical therapy in peripheral facial paresis]. AB - Peripheral facial paresis is a serious clinical and social problem. The effect of physical therapy in 44 patients aged 13-80 was studied. Idiopathic peripheral facial paresis was the cause of mimic muscles dysfunction in 75% of our patients. Medical treatment was conducted according to the established program, consisting in applying combined physical, thermal, electrotherapeutic procedures, exercises and massage. All the patients were examined before physical therapy and after I and II series of procedures, based on Pietruski's evolution table of paresis scored in point and percent scale. Patients between age 13-44 demonstrated 47-50% mimic facial muscles efficiency before the treatment, after I series of procedure it amounted to 80-90% and after the II series to 90-100%, so all the symptoms of paresis disappeared. Similar values were scored by patients aged 45-64 years. Minimal percentage of improvement of mimic facial muscles (57%) was noticed in patients aged 65-80 years The increase of the score was evaluated in the studied group after two stages of treatment. It seems to be best in the younger patients. In the examined group the best improvement evaluated as regression of the symptoms was found in patients with idiopathic peripheral facial paresis. The process of improvement was very slow and not full among patients who additionally suffered from co-existing diseases. Patients with diabetes improved by 57%, with arterial hypertension by 53%, the patients with obesity only by 43% after two cycles of treatment. PMID- 11873611 TI - [Therapeutic potential of interferon beta-1b and related drugs in multiple sclerosis: comparative meta-analysis]. AB - The objective of this current meta-analysis is to determine whether IFN beta-1b is substantially better than other immunomodulating, immunosuppressive or anti inflammatory drugs in modifying the clinical course of relapsing-remitting and secondary progressive multiple sclerosis (RR MS, SP MS). A comparison shows that in 2-year treatment IFN beta-1b, beta-1a s.c., glatiramer acetate, immunoglobulin G i.v. and mitoxantrone i.v. reduced annual relapse rate (45%), increased the proportion of relapse-free patients (34%), decreased the mean level of disability (-0.23 in EDSS) and diminished the fraction of RR MS patients with sustained progression (25%); p < or = 0.05. Azathioprine demonstrated considerable delay in the onset of action and loss of efficacy in the third year of treatment. IFN beta 1b and cyclical pulses of intravenous high dose methylprednisolone decreased the annual relapse rate and slowed the progression in SP MS patients; p < or = 0.04. All drugs significantly reduced the number of new and enlarging brain lesions on GdT1 and PD/T2 images in MRI (mean -68%). Furthermore, IFN beta-1b s.c. and beta 1a s.c. evidently reversed burden of the disease (BOD) on T2 images (median 4.7%); p = 0.001. However, glatiramer acetate, immunoglobulin G i.v., IFN beta-1a i.m., methylprednisolone i.m. and possibly azathiopirine failed to diminish BOD on T2 images; p = ns. It is concluded that in most RR MS patients IFN beta-1b has similar clinical effect to other immunomodulating drugs and mitoxantrone. PMID- 11873612 TI - [Response of the immunological system to infection with particular reference to the role of NF-kappa B agent]. AB - Main components of innate and acquired immunity participating in the response to infection were discussed. The role of NF-kappa B transcription factor was stressed as a key regulatory agent governing the immune response to infection. Attention was also paid to the escape mechanisms of microbes to avoid immune reaction of the host. PMID- 11873613 TI - [Viral infections in the pathogenesis of multiple sclerosis]. AB - In the paper the current opinion on the role of viral infections in the pathogenesis of multiple sclerosis is presented. The results of epidemiological and serological studies are discussed as well as the results of viral isolation attempts and the search for virus structures in CNS of MS patients. Relation between viral infections and MS exacerbations and postulated mechanisms of virus induced demyelination is described. PMID- 11873614 TI - [Treatment of neurodegenerative diseases: new perspectives]. AB - The experimental models of neurodegeneration give a possibility to study the inflammatory reaction that starts in response to neuronal death. Inflammation consists of microglial and astroglial activation, expression of new molecules as cytokines, adhesion molecules and MHC antigens, and is potentially neurotoxic. This article is a summary of a few latest studies that investigate anti inflammatory agents effect on neuron survival in MPTP mice model of Parkinson's disease. Murine model of Parkinson's disease uses a quite selective toxic effect of MPTP on nigrostriatal system. MPTP causes degeneration of dopaminergic cells bodies in the substantia nigra and of their endings in striatum. Our findings show that anti-inflammatory treatment protects neuronal death. It may indicate that the inflammation contributes to the dopaminergic neuron impairment following MPTP intoxication. However this hypothesis needs further investigation because recent studies suggest that inflammation may have also a protective effect in neurodegentration. PMID- 11873615 TI - [Prions, epidemic of Creutzfeldt-Jakob variant disease and global emergency]. AB - We present here the current understanding of "prion" theory and global risk for epidemics of variant Creutzfeldt-Jakob disease (vCJD). Prion is the infectious agent of all transmissible spongiform encephalopaties (TSEs). It is regarded as an aggregate of a pathological conformer (PrPSc) of a normal cellular glycoprotein (PrPC) encoded by a gene, in humans on chromosome 20. The differences between PrPSc and PrPC are largely if not exclusively conformational; PrPC is mostly alpha-helical while PrPSc, beta-pleeted. Furthermore, mutations within the coding sequence (open reading frame) of PrP gene are linked with phenotypic expression of CJD, Gerstmann-Straussler-Scheinker disease (GSS) and fatal familial insomnia (FFI). VCJD is caused by transmission from cattle infected with bovine spongiform encephalopathy (BSE). PrP purified from vCJD affected brains is characterised by the so called fourth type of glycosylation pattern. The neuropathological hallmark of vCJD is the florid plaque, an amyloid plaque surrounded by a corona of spongiform change. VCJD affects mostly young people (range: 13-74 years) and it is clinically distinguishable from sporadic CJD. The extent of epidemics is currently unknown; but its dynamic (102 cases until July 2001) may suggest "the worst" scenario. PMID- 11873616 TI - [Neurotoxoplasmosis]. AB - Toxoplasmosis represents a clinical problem which focuses attention of several specialists due to the multiorgan pathology it induces and difficulties in evaluating the activity of the morbid process. The infection creates a particularly difficult problem in cases when central nervous system is involved, which frequently leads to unfavorable sequelae. Contemporary data have been presented which pertain to the morphology and life cycle of Toxoplasma gondii, sources of the infection and ways through which the protozoon penetrates the host organism. The pathomechanism of disturbances in the course of toxoplasmosis with involvement of central nervous system is described, as related to the stage of invasion and immune responses of cell-mediated or humoral type. The clinical pathology of congenital and acquired toxoplasmosis is presented, including that developing in patients with immune deficiencies. Diagnostic criteria are given, indispensable in evaluating the activity of toxoplasmosis process, based on contemporary laboratory techniques (antigen detection, detection of antibodies to T. gondii antigen, molecular tests, like PCR, and radiological techniques, like CT and MRI studies. The studies are important not only in evaluation of the primary infection and also for appraisal of reactivation of T. gondii invasion in conditions of acquired immunodeficiency (AIDS), in the course of systemic diseases, during immunosuppressive treatment and in patients who without treatment for toxoplasmosis in the first year of life. PMID- 11873617 TI - [Actual clinical and immunological problems of neurosyphilis and neurotuberculosis]. AB - In the past neurosyphilis and tuberculosis of the nervous system were the basic clinical problems in neurology. The decrease of the incidence rate has changed the situation. However, in the last few years, especially in connection with HIV infection, the incidence has even started to increase. In Poland the epidemiological situation is not alarming, but there exists some concern connected with the epidemic of syphilis and tuberculosis in countries near the Polish eastern border. In the laboratory diagnosis of syphilis the highest specificity and sensitivity has the TPHA test. In the therapy till now the best results are obtained with megadoses of penicillin. Early diagnosis of tuberculous meningitis, what is the decisive factor for positive results of the treatment can be obtained by PCR technique for detection of DNA genome of mycobacterium. In the treatment the method of choice is the concomitant use of four antimycobacterial drugs. However, there exists the serious problem of often observed drug resistance of mycobacterium. PMID- 11873618 TI - [Acute transverse myelitis]. AB - Acute transverse myelitis may be caused by many factors, however, in most cases the cause cannot be clinically found, which justifies the diagnosis of "Myelitis transversa acuta" or "myelopathia transversa acuta" in such cases. The disease is inflammatory spinal demyelination, differing morphologically from multiple sclerosis. Magnetic resonance is the examination which discloses the injury of several spinal segments. The upper limit of the lesion is higher than the clinical symptoms indicate. There is protein increase and pleocytosis in the cerebrospinal fluid. In most cases the prognosis is favourable; in 33% of patients complete regression of symptoms takes place; 33% present significant improvement and 33% show permanent disability. The frequency of relapses is high and then multiple sclerosis must be suspected. There also occur cases of monophasic multiple sclerosis and relapses of the disease without other symptoms of multiple sclerosis. The treatment of choice are steroids administered in high doses. PMID- 11873620 TI - [Tick-borne encephalitis versus multiple sclerosis: clinical and immunological aspects]. AB - The condition of making the diagnosis of tick-borne encephalitis is the performance of serological investigation, quite apart from careful evaluation of medical history and clinical status of patient. Contemporary diagnostics of tick borne encephalitis uses immuno-enzymatic test (ELISA), which enables detection of specific IgM antibodies (in initial phase of disease) or IgM and IgG antibodies (in the phase of the disease with neurological symptoms). The highest diagnostic value in the infection phase of disease has the demonstration of at least fourfold increase of viral antibodies level, measured in acute phase and 2 to 6 weeks after that. In case of multiple sclerosis, a laboratory test, specific for the illness, has not been established as yet. The basis for establishing the diagnosis is the clinical picture. Results of auxiliary examinations are used for the completion of clinical evaluation and for differential diagnosis. Seroepidemiological investigations were carried out in multiple sclerosis patients to estimate the level of antibodies against tick-borne encephalitis. The investigations indicated low level of IgG antibodies in patients' serum. PMID- 11873619 TI - [Laryngological experiences in treatment of Bell's palsy]. AB - Between 1990-2000 15 cases of unilateral Bell paresis of the VIIth nerve were treated using local injection of hydrocortisone in the region of the foramen stylomastoideum. Paralelly were administered iontophoresis, galvanisation of the facial mimic muscles together with vasodilating drugs. Early introduced treatment gave complete return of facialis function. The usefulness of treatment monitoring with topodiagnostic tests is stressed. PMID- 11873621 TI - [Acquired inflammatory neuropathies in children and their therapy]. AB - Neuropathies where there is an association with acquired peripheral nerves dysfunction and inflammation include inflammatory neuropathies (IN), as well as sequelae of vaccinations involving peripheral nerves. In a small portion of these diseases central nervous system is involved. In the years 1996-2000, among 22 children with acute flaccid paresis who were hospitalized in the Krakow Department of Paediatric Neurology, there were 16 patients with IN, including 13 with Guillain-Barre syndrome, single cases of Miller-Fisher syndrome, chronic inflammatory demyelinating polyneuropathy involving central nervous system and neuroborreliosis. Additionally, four children were hospitalized for optic neuritis. The author presents data on aetiology, electrophysiology and follow-up of these patients, as well as describes the management and outcome. Apart from their cognitive and practical value, these data significantly correspond with the currently implemented program of poliomyelitis eradication. PMID- 11873622 TI - [Neuronal grafts: a successful connection between basic and clinical research]. PMID- 11873623 TI - [Susceptibility of Pseudomonas aeruginosa to antibiotics isolated from patients of intensive care units in France in 1998. Resistant phenotypes to beta-lactams]. AB - Pseudomonas aeruginosa is responsible for nosocomial infections and demonstrates many types of resistance mechanisms to antibiotics. Thus, in vitro susceptibility survey are frequently required. In this study, susceptibility has been assessed on 105 non redundant consecutive strains isolated from ICU's in 18 general hospitals, from 01.02.98 to 30.06.98. Only clinically significant samples have been considered. MICs have been measured for nine beta-lactams, three aminoglycosides, one fluoroquinolone and colistine. For ticarcilline resistant strains, phenotype has been assessed on Mueller-Hinton medium supplemented with beta-lactamases inhibitor. Transferable beta-lactamases has been identified using pl and PCR. MIC 50 and MIC 90 (mg/L) for beta-lactams are the following (MIC 50- >90): ticarcilline (16-->512), ticarcilline + clavulanic acid (16-->512), piperacilline (4-->512), piperacilline + tazobactam (4-->64), aztreonam (4-->16), cefsulodine (4-->32), ceftazidime (2-->16), cefepime (4-->16), imipeneme (1-->8). For aminoglycosides: gentamicine (2-->32), tobramycine (1-->32), amikacine (4- >16). For ciprofloxacine (0.25-->32) and colistine (0.5-->2). According to CA-SFM break points recommendations, 50% of isolated strains are resistant to gentamicine, one out of three for ticarcilline + clavulanic acid (29%), one out of four for tobramycine (25%) and ciprofloxacine (25%), one out of ten for amikacine (9%), tazocilline (8%) and imipeneme (9%). Resistance to ceftazidime and aztreonam is uncommon (respectively 2%-1%) and never observed for cefepim. For ticarcilline resistant strains, (38% of total isolates) the following phenotypes have been detected: 6.7% non enzymatic resistance, 15.2% transferable beta-lactamase (TEM 4.8%, CARB 4.8%, TEM + CARB 4.8% and OXA-10 and derivated 0.9%) and 16.2% high level cephalosporinase. Extended-spectrum beta-lactamase has never been detected. TEM beta-lactamase is associated with resistance to amikacine and ciprofloxacine. PMID- 11873624 TI - [Clinical and serological characteristics of systemic lupus erythematosus: 128 cases]. AB - OBJECTIVE: To analyse the clinical and serological characteristics of systemic lupus erythematosus (SLE) in the center of Tunisia. METHODS: We studied 128 patients with SLE aged one to 73 years. Antinuclear antibodies (ANA) were detected by an immunofluorescence method. Anti-double-stranded DNA (anti-dsDNA) antibodies, anti-extractable nuclear antigen antibodies (anti-Sm, anti-SS-A, anti SS-B and anti-RNP) and anti-cardiolipin (aCL of IgG, IgA and IgM isotypes) antibodies were detected by ELISA. RESULTS: Malar rash (71%) and anemia (71%) were the most common clinical manifestations. Arthritis was seen in 62.5%. Severe kidney damage was observed in 39%. Pericarditis and pleuritis were observed in only 23%. Neurological manifestations (16%) were uncommon. Clinical manifestations of anti-phospholipid syndrome (SAPL) were observed in 15%. ANA were detected in 100%, anti-dsDNA in 76%, anti-Sm in 55.5%, anti-SS-A in 64%, anti-SS-B in 33.6%, anti-RNP in 49%. aCL of IgG, IgA and IgM isotypes were detected in 63.5%, 49% and 40.6% of the patients respectively. The only significant positive clinical associations were those of arthritis with anti dsDNA antibodies (p = 0.022) and malar rash with anti-SS-A antibodies (p = 0.002). CONCLUSIONS: This study suggests that tunisians with SLE present, in general, a mild form of disease predominantly manifested by cutaneous, musculoskeletal and hematologic involvement but low prevalence of major organ damage. PMID- 11873625 TI - Hispano-Indian admixture in Paraguay studied by analysis of HLA-DRB1 polymorphism. AB - The genetic polymorphism of the Paraguayan population results from the admixture between South American Indians named Guaranis and Spaniards. In order to evaluate the genetic predominance in the Paraguayan population, we typed 50 healthy Paraguayans for HLA-DRB1 by molecular biology and compared their HLA-DRB1 polymorphism to that of the Guaranis and of two Spanish populations. Six significant differences of alleles frequencies were observed between Paraguayans and Guaranis--DRB1*01, 06 (13, 14), 15, 16, 07--whereas only one difference was observed with the Spaniards (DRB1*14). The DRB1*14 frequency was higher in Paraguayan than in the Spanish populations essentially due to the presence of DRB1*1402 related alleles (1402,06,13). These alleles are extremely rare in the Spanish populations whereas frequent in the Guaranis from Brazil and in South American Indian tribes living in the lymitrophe regions of Paraguay (Toba, Wichi and Terena). Thus, the presence of the DRB1*1402 related alleles (6%) in the Paraguayan population constitutes the major Indian contribution to the HLA-DR polymorphism of the Paraguayan population. The genetic distances between Paraguayans and the two Spanish populations were closer (.494 and .415) than that between Paraguayans and Guaranis (.958). Altogether these results suggest the predominance of the Spanish genetic in the Paraguayan population. Historical events are discussed to explain this predominance. PMID- 11873627 TI - Is the incidence of Balkan endemic nephropathy decreasing? AB - Balkan Endemic Nephropathy (BEN) is a non-inflammatory, slowly progressing, familial, primarily tubulo-interstitial, bilateral renal disease that affects rural populations in several Balkan countries. Our study describes a time trend of the incidence of BEN in eight villages of Vratza District, Bulgaria, for the period 1965-1987, based on three various data sets. The data suggest that after the initial peak between 1967 and 1970, the incidence remained quite stable for the period 1970-1984, and declined after 1984. However, the study also demonstrates under-recording of BEN cases and less complete case identification, especially after 1979. Migration of population might also have contributed to an apparent decline in registered cases. We detected cases of BEN in villages that previously were BEN-free. We recommended a rigorous monitoring of BEN in all afflicted countries, before concluding that the incidence of BEN is decreasing. PMID- 11873628 TI - [Cellular transplantation in the treatmnt of ishemic heart failure]. PMID- 11873626 TI - [Evolution of susceptibility of aerobic gram-negative aerobic bacilli to quinolones and fluoroquinolones in a university hospital (1992-2000)]. AB - Susceptibility to quinolones of aerobic gram-negative bacilli was assessed in a 2000-bed university hospital from 1992 to 2000. There was a significant downward trend in the rate of susceptibility to nalidixic acid (Nal) for Enterobacteriaceae as a whole from 1992 to 2000 (86% vs 82%), and E. coli (92% vs 84%), and an upward trend for K. pneumoniae (74% vs 82%), the latter being related to the control of the spread of epidemic ESBL producing strains. The overall susceptibility of Enterobacteriaceae to ciprofloxacin (Cip) paralleled the susceptibility to Nal: decreased susceptibility for Enterobacteriaceae as a whole (96% vs 89%) and E. coli (99% vs 91%). A clear decrease in the level of susceptibility to Cip occurred during the study period among the Nal-resistant strains as demonstrated by the decrease in the median zone diameter (D) observed among the Nal-resistant strains of E. coli (26 mm in 1992 vs 19 mm in 1998-2000). The zone diameter distribution pattern changed from an unimodal distribution in 1992 to a trimodal distribution in 2000 secondary to the occurrence of a population of resistant strains (D = 13 mm) and of a highly resistant population (D = 6 mm). Finally, the susceptibility to Cip of P. aeruginosa strains remained stable around 62% throughout the study period. PMID- 11873629 TI - [Allogenic stem cell transplantation after non-myeloablative conditioning regimen (mini-allogenic" stem cell transplantation)]. AB - Allogeneic stem cell transplantation is able to cure many hematologic malignancies, through, at least partially, a graft versus disease effect of the donor's immune system transfer. However, the toxicity of this technique limits its use to selected patients. The aim of non-myeloablative stem cell transplantation is to reduce the toxicity of the conditioning regimen while allowing the engrafement of donor's stem cells and the immunological antitumoral activity of the donor's immune system. Several reports have already demonstrated the validity of this concept. This new multi-step therapeutic strategy is complex, raises many questions and deserves further studies to be fully applicable to a greater number of patients. PMID- 11873630 TI - [HLA-G: immune tolerance in normal and pathological physiology]. AB - HLA-G is a low polymorphic non-classical major histocompatibility complex class I molecule which can be expressed upon seven isoforms, and whose expression is correlated to immune tolerance situations. Indeed, the expression of HLA-G may allow partially or non histocompatible cells to escape host's immune aimed to eliminate these "foreign" cells. Functions share by HLA-G molecules are the ability to inhibit lysis mediated by natural killer (NK) and by cytotoxic T cells, and to induce apoptosis of activated CD8+ T cells via the HLA-G soluble form. Such model which evades defenses of the immune system was described in the foeto-maternal symbiosis, in tumoral cells or histo-incompatible graft previously. The importance of the role of HLA-G molecules expressed by particular cells as a shield against immune defenses is actually largely studied. The control of HLA-G expression could be used as immunotherapeutic approach, either by up-modulating HLA-G expression in order to favour tolerance of incompatible cells such as foetal cell or graft, or by down-modulating HLA-G expression in order to support the destruction of tumoral cells by cytotoxic T cells. This article reports the recent data on polymorphism, HLA-G gene transcription and implication of HLA-G molecules in immune responses during pregnancy, transplantation or tumoral growth. PMID- 11873631 TI - [Development of nutritional obesity in transgenic mice with an adrenergic receptivity in adipose tissue comparable with that of humans]. AB - Obesity is characterized by an excessive development of fat mass which is a consequence of increased fat cell size and/or fat cell number. Several hormones and neurotransmitters are regulators of adipose tissue development and metabolism. Among them, catecholamines play a major role by acting through alpha 2- and beta-adrenergic receptors. Stimulation of alpha 2-adrenergic receptors induce inhibition of lipolysis in mature adipocytes as well as preadipocyte proliferation. The antilipolytic effect mediated by alpha 2-adrenergic receptors is in part responsible for the weak lipid mobilization of some fat deposits in humans (subcutaneous fat in particular). Changes in beta- and alpha 2-adrenergic receptors ratio and function have been proposed to explain the lipolytic disturbances described in some obese subjects. Human and rodent adipocytes differ considerably with respect to the balance between beta- and alpha 2-adrenergic receptors. Human adipocytes express mainly alpha 2- but very few beta 3 adrenergic receptors while the reverse is true for rodent adipocytes. Since no suitable animal model was available to study the contribution of alpha 2/beta adrenergic balance in adipocytes in vivo, we combined gene targeting and transgenic approaches to create a mice with increased alpha 2/beta-adrenergic ratio in adipose tissue. Specifically, we have generated transgenic mice strains on a beta 3-adrenergic receptor knock-out background which express human alpha 2 adrenergic receptors. No particular phenotype was observed in mice maintained in normal diet whereas when fed a high fat diet, transgenic mice increased significantly body weight and fat mass. These results underline the physiologic relevance of the interaction of the presence of alpha 2-adrenergic receptors with a high fat diet in the control of adipose tissue development. PMID- 11873632 TI - [in vitro activity of telithromycin against Haemophilus influenzae]. AB - The aim of this study was to evaluate the in vitro activity of telithromycin against 142 strains of Haemophilus influenzae using determination of MICs by agar dilution method and to evaluate the correlation between MICs and inhibition diameter zones obtained by disk diffusion testing. MIC50 and MIC90 of telithromycin were 1 and 2 mg/L respectively. Telithromycin activity against H. influenzae was similar to that of azithromycin, superior to erythromycin and clarithromycin and irrespective of the susceptibility to betalactams. Distribution of diameter zones showed a similar pattern to that of MICs but correlation between MICs and diameter zones was poor with correlation coefficients inferior to 0.5 whatever the agar media used. PMID- 11873633 TI - [Identification of plasmid-encoded cephalosporinase ACC-1 among various enterobacteria (Klebsiella pneumoniae, Proteus mirabilis, Salmonella) isolated from a Tunisian hospital (Sfax 997-2000)]. AB - Because a multiresistant K. pneumoniae outbreak detected in an intensive care unit of a parisian hospital, combined to the production of the plasmid-encoded cephalosporinase ACC-1, a probable importation via a patient was suggested from another country (Tunisia). The investigation was conducted to examine 35 clinical strains of enterobacteria resistant to ceftazidime without synergy towards Augmentin. Other test of synergy with two inhibitors, BRL 42715, Ro 48-5545 was performed by diffusion method and deposit of 10 micrograms of inhibitor on disks containing ceftazidime, cefoxitin and cefotetan. Synergies were obtained suggesting a probable production of ACC-1 type among six isolates of K. pneumoniae (two), Proteus mirabilis (one) and Salmonella (three) issued from different units. The isoelectric focusing on gel revealed at least one band of beta-lactamase activity at 7.8 but also demonstrated the simultaneous production of several probable beta-lactamases including TEM-type, SHV-2 and ACC-1 among S. enterica ser. Livingstone. The PCR of the gene blaacc-1 was positive. The sequencing (1160 pb) of two products showed high identity (99-100%) with the gene blaacc-1 deposited in 1999. Finally the ACC-1 type reported in Tunisia was probably imported in France via a patient. Because a simultaneous synthesis of ESBL and ACC-1 type, its presence may be invisible and need more investigation. PMID- 11873634 TI - [New intraoperative imaging technologies]. PMID- 11873635 TI - [Current role of traditional endoscopy and prospects for virtual colonoscopy in the management of colorectal neoplastic lesions]. AB - The Authors, reviewing their surgical experience with colo-rectal cancer in the last 13 years, conclude that, in the management of this condition, the routine use of traditional endoscopy will continue to have a crucial role in terms of prevention, cure and/or survival. It is possible that in the future present limits of endoscopic techniques will be seperated by routine use of endoscopic ultrasounds and virtual colonoscopy. PMID- 11873636 TI - [Upper gastrointestinal hemorrhage of unusual etiology: clinical case and technical note]. AB - The Authors report a case of upper digestive tract haemorrhage at atypical side, the diagnosis being often difficult as well as therapeutical options, particularly referent to the surgical solutions adopted. PMID- 11873637 TI - [Gastrointestinal stromal tumors: a case with mixed muscular-neuronal origin]. AB - GIST have still today controversial aspects of their histogenesis that are reflected on the classification and prognosis. It's showed a particular case with mixed muscular-neuronal histogenesis that supposes totipotent cell origin of these neoplasms. Diagnosis, classification and prognosis are very difficult to evaluate; surgery at the moment is the only useful treatment modality. PMID- 11873638 TI - [Ultrasound-guided trans-parietohepatic cholecystostomy in the critical patient: current indications]. AB - The appearance of acute cholecystitis can make to complicate a natural history of cholelitiasis or post-operating time of patients that have concomitant predisposition factors. The best therapy is the cholecystectomy but somewhere for the critical general conditions is too much dangerous to make a surgical procedure. However we need to stabilize patients conditions, also for a short time. Our experience suggest us that percutaneous transhepatic cholecystostomy is a simple method without any complications, efficacious to resolve the acute sepsis in patients with cholecystitis that not be able to tolerate a surgical procedure. PMID- 11873639 TI - [Randomized prospective study of early removal of drainage in breast cancer surgery]. AB - Axillary seroma is absolutely the most frequent complication of breast cancer surgery. The Authors have accrued 100 consecutive breast cancer patients in a randomized study in order to compare seroma incidence by removing drains on 2nd postoperative day (1st arm) versus 3rd postoperative day (2nd arm); 48 patients were accrued in the first arm and 52 in the second. All patients received a standard axillary dissection. Two suction drains were placed. A compressive medication was applied after surgery. Patients started physiotherapy on the 1st postoperative day. The overall seroma prevalence was 21%. We have 8/48 (16%) seromas in the 1st group and 13/52 (25%) in the 2nd. No significant differences were registered between two arms. Clinical seroma was treated by needle aspiration and medication with a steroid. Conclusions coming out from this study are: 1) early drains removal doesn't increase seroma rate; 2) axillary clearance has to be performed removing en bloc the fatty tissue respecting surgical plains; 3) apply a compressive bandaging; 4) early arm physiotherapy; 5) medication with steroid may reduce the fluid formation. PMID- 11873640 TI - [New perspectives in the treatment of liver metastasis from colorectal cancer: radiofrequency thermal ablation]. AB - Preliminary results of new treatment of metastatic hepatic malignancies from colo rectal cancer, the radiofrequency ablation (RFA), are reported. The method is limited by few cases treated and short follow-up but it opens new perspectives in metasurgical treatment of these lesions as regards the previous experiences based on wide numbers of patients, recently reported in the literature. PMID- 11873642 TI - [Deep venous thrombosis after leg amputation. Review of the argument]. AB - Patients undergoing lower extremity amputation are perceived to be at high risk for deep vein thrombosis (DVT). DVT can cause micro or macro pulmonary embolism and often the post-thrombophlebitic syndrome. The chronic condition can affect patient quality of life and his residual working capacity. Usually the echo Doppler or the color-Doppler is used as a prevention and diagnostic method, identifying patients at high risk. Following the Authors examine and report the Literature opinion about the topics. PMID- 11873641 TI - [Current role of endoscopic ultrasonography in the treatment of colorectal neoplastic lesions]. AB - The Authors examined the value of endoscopic ultrasounds in the tmanagement of colo-rectal lesions, particularly neoplastic ones, on the basis of their surgical experience, concluding that to date such technique cannot substitute traditional endoscopy, but surely can be a valid complementary tool for imaging diagnostics, expecially to study wall tumoral infiltration or extra-parietal relapses. PMID- 11873643 TI - Mandatory patient safety training, from the board room to linen room. AB - Here is a list of counter-intuitive statements about patient safety, a list of required reading on the subject, and an excerpt of a tool that can help you analyze your organizations patient safety strengths and weaknesses. PMID- 11873644 TI - Natural disaster reaction averts a Houston disaster. AB - Having a disaster plan is one thing. But how will it work in a real disaster? One Houston hospital found out last June. PMID- 11873645 TI - Proper comparisons are key to hospital benchmarking. PMID- 11873647 TI - Patient Safety Alert. HHS commits $50 million to better patient safety. PMID- 11873646 TI - Patient Safety Alert. Anthrax outbreak forces closer focus on patient safety. PMID- 11873648 TI - Production of chimeras derived from murine embryonic stem cells. PMID- 11873649 TI - Gene-targeting strategies. PMID- 11873650 TI - Cre/loxP recombination system and gene targeting. PMID- 11873651 TI - Cryopreservation of transgenic mouse lines. AB - Cryopreservation will, if used appropriately, lower maintenance costs while creating embryo or germline banks from which transgenic lines can be restored. To maximize the likelihood of restoring a transgenic line, it is best to establish that the gametes or zygotes of that strain do survive cryopreservation before all of the animals have been culled. Furthermore, it is advisable to store more than one type of material (e.g., sperm and embryos) and to ensure that the material is stored under optimal conditions (i.e., at low subzero temperatures) and never partially or wholly thawed, until it is removed for use. PMID- 11873652 TI - Ovarian tissue transplantation and cryopreservation. Application to maintenance and recovery of transgenic and inbred mouse lines. PMID- 11873653 TI - Transgenesis in the rat. PMID- 11873654 TI - Gene transfer in Drosophila. PMID- 11873655 TI - Generation of transgenic livestock by pronuclear injection. PMID- 11873656 TI - Transgenic sheep from cultured cells. PMID- 11873657 TI - Biomedical and agricultural applications of animal transgenesis. PMID- 11873658 TI - Analysis of transgenic mice. PMID- 11873659 TI - Oocyte injection in the mouse. PMID- 11873660 TI - Adenoviral infection. PMID- 11873661 TI - Retroviral infection. PMID- 11873662 TI - In vitro isolation of murine embryonic stem cells. PMID- 11873663 TI - [Multimodal treatment of hepatocellular carcinoma]. AB - BACKGROUND: Several options now available create the need for a multidisciplinary approach to the treatment of patients with hepatocellular carcinoma (HCC). OBJECTIVE: To describe the experience with a group of patients with HCC diagnosed and treated with the current modalities for control of this entity. POPULATION AND METHOD: A total of 169 consecutive patients with HCC treated between January 1990 and February 1998; 41 liver resections, 14 liver transplants, 87 chemoembolizations and 8 alcoholizations were performed. The median follow-up time was 23.6 months (range: 0.2-94). In the other 29 patients, only support treatment was administered. RESULTS: Resected patients had a lower median hospitalization rate, blood transfusions, and hospitalization mortality. Recurrence was 42.1% in the resected group but 0% in transplanted patients. The overall survival at 36 months was better for transplanted than for resected patients (100% vs. 62.3%; p < 0.02). The median survival time was 13 months (IC: 95%: 11-15 months) in chemoembolizated patients. CONCLUSIONS: 1. The outcome of patients with HCC is influenced by several factors, therefore calling for multimodal approach. 2. Surgical procedures should be indicated with low morbimortality rates. 3. The overall survival and disease free survival time is better in transplanted patients than in the resected group. 4. The survival time of patients with unresectable HCC without extra-liver disease could be improved with chemoembolization. PMID- 11873664 TI - [Enteroendocrine cells modifications in Helicobacter pylori gastritis]. AB - OBJECTIVE: The aim of the present study was to analyze the modifications in the number and distribution of enteroendocrine cells (EEC) in antrum of patients with Helicobacter pylori (HE) gastritis. We also wanted to demonstrate their possible participation in the immune response. MATERIAL AND METHODS: Twenty-six (26) biopsies of gastric antrum from patients between the ages of 40 and 60 were used. Slides were stained with H&E, Giemsa for HP, and chromogranin A to visualize EEC. Five (5) patients were normal controls. Eleven (11) patients had antral chronic gastritis (ACG) with different grades of activity, and ten (10) patients had multifocal atrophic gastritis (MAG), both groups associated to HP. EEC were quantified in relation to 100 epithelial cells. Results were statistically compared. RESULTS: In the normal control group, EEC were sparsely distributed, deep in antral glands, with an average 19.51 EEC/100 epithelial cells. In ACG there were 12.01/100. Besides EEC were irregularly distributed, close to inflammatory areas, or near lymphoid follicles. CONCLUSION: The decrease in EEC is probably due to degranulation and later to a disappearance or inhibition of stem cells by inflammatory products in HP gastritis. The proximity of EEC to prominent inflammatory zones may indicate EEC modulate the immune response. They produce and excrete peptides that interact with membrane receptors found in T lymphocytes and macrophages. PMID- 11873665 TI - [Acute liver failure: clinical-epidemiological characteristics]. AB - In order to ascertain the clinical and epidemiological features of acute liver failure (ALF), we analyzed the clinical histories of 22 patients from La Plata city, with the diagnosis of ALF (prothrombin level or factor V below 50%) seen between November 1996 and November 2000. Age, sex, hepatic encephalopathy, reason for consultation, etiology, hepatic biochemical tests, serum creatinine, glycemia, digestive hemorrhage, course and treatment variables were analyzed. What is remarkable is the high frequency of the toxic etiology and of infection by HDV, as well as the high prevalence of ascites and the low incidence of hepatic encephalopathy. We think that the high survival rate we found is due to the early diagnosis and early referral of the patients to the intensive care unit and to centers with programs for liver transplantation. PMID- 11873666 TI - [Acute experimental pancreatitis in the opossum. The role of prior truncular vagotomy on the inflammatory response]. AB - BACKGROUND: Acute pancreatitis (AP) is a common cause of admission to the emergency room. Its etiopathogeny is poorly understood. The pancreatic inflammatory response in this process is unclear. The influence of the autonomic nervous system is a controversial issue. AIMS: To demonstrate the effects of truncular vagotomy on AP due to duodenal distention in the South American opossum. PLACE OF APPLICATION: Department of experimental surgery (Churruca-Visca Hospital) INEUCI (Neuroscience Institute, CONICET, UBA). POPULATION: Male and female South American opossum divided into: a control group (7 animals); group A: truncular vagotomy and pyloroplasty seven days after the onset of AP (7 animals). Group B: truncular vagotomy and pyloroplasty 30 days after provoking AP (7 animals). Group C: truncular vagotomy and pyloroplasty 45 days after the onset of AP (7 animals). METHODS: Acute pancreatitis was caused by duodenal distention of the second portion by inserting a Foley catheter through a gastrostomy. CONCLUSIONS: The method of provoking AP is original. The influence of autonomous nervous system is being underestimated in most of the literature available. PMID- 11873667 TI - [Pancreatic tumor: an unusual presentation of an occult breast carcinoma]. AB - A case of cholestasis due to a synchronous pancreatic head metastasis from an occult lobular breast carcinoma is presented. The patient had a clinical and radiological picture compatible with a pancreatic head primary tumor with cholestasis and ascites. Ultrasonographically guided fine needle aspiration cytology demonstrated a metastatic breast lobular cancer (positive for cytokeratin AE1 and AE3, cytokeratin 7 and epithelial membrane antigen and negative for cytokeratin 20, CA 19.9, CA 125, CEA and estrogenic receptors). The same cytologic findings were observed in skin and subcutaneous armpit nodules. Clinical and radiological breast examination was unable to demonstrate any tumor in the breast. Pancreatic metastases are rare events and the majority of them are secondary to renal and lung cancer and rarely to breast cancer. In these latter cases, metastases are usually disclosed after a disease-free interval of months or years between primary tumor resection and recognition of the pancreatic tumor. Synchronous presentation is extremely rare. Metastases of epithelial origin are uncommon in pancreas and generally are first misdiagnosed as primary pancreatic cancer. Fine needle aspiration is a useful tool for the differential diagnosis in patients with widespread disease. PMID- 11873668 TI - [Disseminated infection due to strongyloides stercoralis in AIDS patients. A report of 2 cases]. AB - Strongyloides stercoralis is an intestinal nematode that infects humans worldwide. Infected patients with severe involvement of cellular immunity may develop a syndrome characterized by the dissemination of larvae throughout the body. Extraintestinal strongyloidiasis has been infrequently reported and despite the prevalence of the helminth in tropical and developing countries there are few cases reported in AIDS patients. Most patients with disseminated strongyloidiasis present with fever, cough, diarrhea and shortness of breath. Chest radiographs usually show diffuse infiltrates. The diagnosis has been made by finding the helminth in respiratory secretions or stool. Enteric organisms like Escherichia coli can often be isolated in the blood or cerebrospinal fluid. We report two cases of disseminated strongyloidiasis in AIDS patients, in which stercoralis larvae were detected in sputum and stool samples. PMID- 11873670 TI - [Acute bacterial hepatitis]. PMID- 11873669 TI - Primary cancer of the cystic duct; a case report. AB - Cancer of the cystic duct is very infrequent, the diagnosis must be made during surgery for biliary pathology, generally lithiasis. The present tendency as regards treatment, is surgery associated with lymphadenectomy. PMID- 11873671 TI - [Focal atrophy with stellate scars in children with chronic gastritis associated to Helicobacter pylori. Is this early atrophic gastritis?]. AB - BACKGROUND: Atrophic gastritis has not been described in children in the setting of Helicobacter Pylori infection. METHODS: Gastric biopsies of six children (7 to 11 years old) with history of HpCG and recent therapeutic eradication of H. Pylori, were reviewed. In the 6 H. Pylori was documented with histology, culture, direct visualization and/or serology before treatment. Cases were compared with five biopsies of age-matched patients showing none of the above-mentioned clinical data. All the biopsies were formalin-fixed, paraffin embedded and stained with hematoxilin-eosin, Masson trichrome and reticulin stain. RESULTS: The biopsies of the six treated patients showed variable-in-size stellate-shaped spots of glandular loss replaced by dense connective tissue with few inflammatory cells. The fibrous tissue showed a central area of scarring and radially oriented spikes extending to adjacent interglandular tissue, more evident with the Masson trichrome stain. Density of inflammatory cells in the lamina propria was variable. H. Pylori organisms were consistently absent. On the reticulin stain the atrophic areas showed coarser and compacted reticulin. Stellate scars were not present in the five controls. CONCLUSIONS: Small foci with fibrous scars may be found in children with long standing HpCG, perhaps as an early sequel of it. We hypothesize that if the chronic gastritis-gastric atrophy process is a continuum, these stellate scars may be representing the very beginning of the multifocal atrophic gastritis usually seen in adult patients. PMID- 11873672 TI - [Acute hepatic failure. A syndrome with a not definitive delimitation but with better prognosis]. PMID- 11873673 TI - School dysfunction: evaluative aspects. AB - Inability of a child to function in a formal school setting necessitates a review of potential physical, mental, emotional, and behavioral etiologies. A thorough, systematic history and examination is expected; the time of onset, age, and grade of the patient helps to narrow the differential diagnosis. A careful assessment should determine the work-up, correct diagnosis, and treatment. Additional primary care physician responsibilities in school dysfunction cases include patient education, family support, and specialist referral, if necessary. PMID- 11873674 TI - Pulmonary carcinosarcoma. AB - Pulmonary carcinosarcoma is a rare malignancy, comprising less than 0.3% of all malignant neoplasms of the lung. Since the first report of this tumor by Kika in 1908, approximately 100 cases have been described in the literature. In this report, we present five cases treated by our group over a period of 20 years. PMID- 11873675 TI - Mediation of patient complaints. A challenge in 1990--a reality in 2002. PMID- 11873676 TI - A model of extraordinary social engagement, or "moral giftedness". PMID- 11873677 TI - The development of an enduring commitment to service work. PMID- 11873678 TI - Predictors of positive cooperative behavior in youths. PMID- 11873679 TI - The nature of vital engagement in adulthood. PMID- 11873680 TI - Encouraging mentorship in young scientists. PMID- 11873681 TI - Continuing motivation beyond the high school classroom. PMID- 11873682 TI - Multidisciplinary cleft management. PMID- 11873683 TI - Primary repair of cleft palate: a review of surgical technique. PMID- 11873684 TI - Secondary bone grafting for unilateral alveolar clefts: a review of surgical techniques. AB - For the past 5 years we have been performing the closure of the oronasal fistula with alveolar bone grafting in the early-to-late secondary time period. The final decision as to whether early or late grafting should be done is based on if a grafting is needed to support the eruption of a functional lateral incisor. Generally the Moczair type buccal flap, with a "Z" release for wide clefts, is used for the buccal flap. This is combined with lateral releasing incisions on the palate for palatal closure. Separate nasal and oral closures are performed in all cases, with bone placed between the two layers in the alveolar defect. It is believed that this treatment sequence best fulfills the criteria for successful alveolar bone grafting outlined at the beginning of this chapter. Figure 8 demonstrates an alveolar bone grafting procedure in a 10-year-old girl just before eruption of the canine tooth treated with a buccal Moczair flap, lateral releasing incisions on the palate, two-layered closure, and the placement of an iliac bone graft. PMID- 11873685 TI - Bilateral cleft alveolus surgery. PMID- 11873686 TI - Primary repair of the unilateral cleft lip. PMID- 11873687 TI - Orthognathic surgery in the patient with cleft lip and palate. PMID- 11873688 TI - Maxillary intraoral reconstruction with regional flaps. PMID- 11873689 TI - Arthrocentesis. PMID- 11873690 TI - Associated bony procedures for preservation. PMID- 11873691 TI - Surgical correction of temporomandibular dislocation. PMID- 11873692 TI - Temporomandibular joint disc preservation. PMID- 11873693 TI - Use of local tissues for temporomandibular joint surgery disc replacement. AB - The CTM flap offers the oral and maxillofacial surgeon an excellent graft material that can be used to replace, anatomically and functionally, a nonrestorable TMJ disc or failed alloplastic implant. It has numerous advantages over the existing autogenous, allogeneic, and alloplastic materials currently available. In addition, the anatomic location of the flap makes the logistics of harvesting the graft much easier. PMID- 11873695 TI - The condylotomy procedure. PMID- 11873694 TI - Autogenous dermal and auricular cartilage grafts for temporomandibular joint repair. PMID- 11873696 TI - Consultation, surgical anatomy, and esthetic nasal evaluation. PMID- 11873698 TI - Functional septorhinoplasty. PMID- 11873697 TI - Fundamental terms, considerations, and approaches in rhinoplasty. AB - It is essential to appreciate the healing dynamics in rhinoplasty patients that occurs over a period of months. It is for this reason that long-term analysis and follow-up are essential for the surgeon to fully realize and understand the final effects of the effort. Discussion of important patient selection considerations, review of standard terminology, review of incision and approach choices, and brief review of alar cartilage procedures were presented. The importance of performing minimal surgery that would be capable of predictably producing the desired changes is stressed. In the majority of rhinoplasty cases, use of nondelivery or limited-delivery approaches, limited transfixion incisions, and conservative alar rim strip techniques seems to offer maximal predictability. These considerations, however, must be balanced with the improved exposure and teaching opportunity allowed via open rhinoplasty. The decision of which approach to use must be based on a thorough understanding of the problems presented by each case, and the techniques available within a surgeon's armamentarium. Excellent exposure and control of symmetry in the open structure approaches has led many teaching institutions to rely solely on that approach. It is the author's opinion that nasal reconstruction in more complicated secondary rhinoplasty, severely distorted, or congenitally deforms noses, or complex combinations of anatomic structure requiring grafting or lengthening should be performed via open rhinoplasty. It is believed, however, that an effective rhinoplastic surgeon should be able to successfully accomplish most nasal correction via any of the standard approaches discussed. With more conservative nondelivery approaches comes the probability of reduction in surgical trauma, decreased scarring, and increased relative predictability necessary for cosmetic surgical cases. Unfortunately, limitations of exposure and access are often difficult for even very experienced surgeons. It is not acceptable that all patients have to be exposed to only one type of exposure or technique based solely on the individual's original training experience. Rhinoplasty offers an ongoing learning experience, with a variety of elements. It remains perhaps the most challenging and demanding of the procedures in cosmetic and reconstructive surgery. PMID- 11873699 TI - Augmentation rhinoplasty. AB - The combination of grafts and skeletal maneuvers will usually yield a nose that is esthetic and functional. Subtleness is the key--the nose should neither be stark in appearance nor impart the sensation of rigidity to feel. Where multiple grafts are called for, they are preferably nonarticulated so as to give the sensation of natural "give" to the touch, and forbearance against trauma as in the native nose. Constructive and reconstructive tactics for the nose, while based in sound rhinoplastic and craniomaxillofacial protocols, often include judicious usage of augmentative materials and techniques. For both appearance and also for function, their use is most gratifying for the patient and for the clinician. PMID- 11873700 TI - Surgery of the nasal tip. PMID- 11873701 TI - Surgical management of the alar base. PMID- 11873702 TI - Revisional rhinoplasty. PMID- 11873704 TI - September 2001--defining the 21st century. PMID- 11873703 TI - Rhinoplasty. Introduction. PMID- 11873705 TI - Schizophrenia: unravelling treatment. PMID- 11873706 TI - Atypical antipsychotics: mechanism of action. AB - BACKGROUND: Although the principal brain target that all antipsychotic drugs attach to is the dopamine D2 receptor, traditional or typical antipsychotics, by attaching to it, induce extrapyramidal signs and symptoms (EPS). They also, by binding to the D2 receptor, elevate serum prolactin. Atypical antipsychotics given in dosages within the clinically effective range do not bring about these adverse clinical effects. To understand how these drugs work, it is important to examine the atypical antipsychotics' mechanism of action and how it differs from that of the more typical drugs. METHOD: This review analyzes the affinities, the occupancies, and the dissociation time-course of various antipsychotics at dopamine D2 receptors and at serotonin (5-HT) receptors, both in the test tube and in live patients. RESULTS: Of the 31 antipsychotics examined, the older traditional antipsychotics such as trifluperazine, pimozide, chlorpromazine, fluphenazine, haloperidol, and flupenthixol bind more tightly than dopamine itself to the dopamine D2 receptor, with dissociation constants that are lower than that for dopamine. The newer, atypical antipsychotics such as quetiapine, remoxipride, clozapine, olanzapine, sertindole, ziprasidone, and amisulpride all bind more loosely than dopamine to the dopamine D2 receptor and have dissociation constants higher than that for dopamine. These tight and loose binding data agree with the rates of antipsychotic dissociation from the human-cloned D2 receptor. For instance, radioactive haloperidol, chlorpromazine, and raclopride all dissociate very slowly over a 30-minute time span, while radioactive quetiapine, clozapine, remoxipride, and amisulpride dissociate rapidly, in less than 60 seconds. These data also match clinical brain-imaging findings that show haloperidol remaining constantly bound to D2 in humans undergoing 2 positron emission tomography (PET) scans 24 hours apart. Conversely, the occupation of D2 by clozapine or quetiapine has mostly disappeared after 24 hours. CONCLUSION: Atypicals clinically help patients by transiently occupying D2 receptors and then rapidly dissociating to allow normal dopamine neurotransmission. This keeps prolactin levels normal, spares cognition, and obviates EPS. One theory of atypicality is that the newer drugs block 5-HT2A receptors at the same time as they block dopamine receptors and that, somehow, this serotonin-dopamine balance confers atypicality. This, however, is not borne out by the results. While 5-HT2A receptors are readily blocked at low dosages of most atypical antipsychotic drugs (with the important exceptions of remoxipride and amisulpride, neither of which is available for use in Canada) the dosages at which this happens are below those needed to alleviate psychosis. In fact, the antipsychotic threshold occupancy of D2 for antipsychotic action remains at about 65% for both typical and atypical antipsychotic drugs, regardless of whether 5-HT2A receptors are blocked or not. At the same time, the antipsychotic threshold occupancy of D2 for eliciting EPS remains at about 80% for both typical and atypical antipsychotics, regardless of the occupancy of 5-HT2A receptors. RELEVANCE: The "fast-off-D2" theory, on the other hand, predicts which antipsychotic compounds will or will not produce EPS and hyperprolactinemia and which compounds present a relatively low risk for tardive dyskinesia. This theory also explains why L-dopa psychosis responds to low atypical antipsychotic dosages, and it suggests various individualized treatment strategies. PMID- 11873707 TI - Cognitive therapy for schizophrenia: from conceptualization to intervention. AB - OBJECTIVES: To outline the cognitive understanding of symptoms of schizophrenia, such as delusions, hallucinations, and emotional withdrawal, and to review the cognitive therapy approach to ameliorating these symptoms. METHOD: We identified studies examining cognitive factors associated with symptoms of schizophrenia by electronic search (using Medline and Psycinfo). This paper integrates experimental findings and clinical treatment. RESULTS: Recent studies focusing on the psychological aspects of schizophrenia demonstrate the importance of common cognitive biases and distortions that are functionally related to the maintenance of symptoms. Understanding the disorder in cognitive terms provides a framework for psychotherapeutic intervention. Adapting cognitive strategies successfully used in cognitive therapy of depression and anxiety provides an important adjunct to standard treatment of schizophrenia. CONCLUSION: Given that the outcome of current treatment for schizophrenia remains poor, attention to therapist training in psychological approaches is essential. PMID- 11873708 TI - Alternatives to acute hospital psychiatric care in east-end Montreal. AB - OBJECTIVE: As pressure mounts to reduce the number of costly acute care beds, governments and the literature propose top-down ratios. Is this reasonable and fair to the responsible medical officers who, as the key care providers, will need to admit patients and develop discharge plans in a reduced-beds environment? METHOD: Treating physicians of all acute care inpatients on a given day (n = 212) and all new acute care admissions over a 2-week period (n = 125) completed an adapted version of the Nottingham Acute Beds Use Survey (NABUS) Questionnaire. RESULTS: On a given day, only 62 of 212 inpatients were unsuited for any alternative to acute care hospitalization. A floor ratio of 18 acute care beds per 100,000 inhabitants seems adequate for the catchment area in question, provided that alternatives to hospitalization are fully and efficiently available. Alternatives essentially involve an array of the following: supervised residential settings, day hospitals, and intensive home care (2 to 6 hours weekly). The ratio of intensive home care workers required would be 25 per 100,000 inhabitants. PMID- 11873709 TI - A follow-up study of patients with DSM-IV schizophreniform disorder. AB - BACKGROUND: Schizophreniform disorder (SFD) has an unclear diagnostic and prognostic status within the psychotic spectrum. METHOD: We studied 36 inpatients admitted to our ward between 1983 and 1993 due to SFD. The patients were contacted an average of 12 years after index hospitalization, and we noted the course of their illness, as well as their present diagnosis. RESULTS: Of the sample, 84% had additional, mostly psychotic, episodes during the follow-up, and 70% had diagnoses in the schizophrenic spectrum (that is, schizophrenia and schizoaffective disorder). A survival analysis revealed that confusion and the presence of at least 2 good prognostic factors (GPF) at index hospitalization predicted better outcome. CONCLUSIONS: SFD seems to be an early manifestation of schizophrenia. Only a few of those sampled did not experience additional relapses -a pessimistic finding at 12-year follow-up. The findings of this study accord with DSM-IV criteria and the literature regarding the long-term prognosis of SFD and the importance of the GPF. PMID- 11873710 TI - [The translation of questionnaires and of tests: techniques and problems]. AB - OBJECTIVES: To summarize the difficulties involved in translating tests, to describe the translation methods and the test validation procedures, and to apply those to a personality test. METHOD: The revised Freiburg Personality Inventory (FPI-R) was translated, then subjected to the following test validation methods: backtranslation, pretest, and review by a carefully selected expert committee. RESULTS: We used a literature review to clarify FPI-R translation problems. These include in particular the different types of equivalence between the source language and the target language (for example, semantics and idioms, as well as experiential and conceptual equivalence). Statistical validation procedures are employed in principle only. CONCLUSION: The current method combining translation with backtranslation is not sufficient and must be used with, at least, a pretest and step-by-step review by an expert committee. The presence of unilingual experts to explain the smallest details of the target language, which bilingual experts could miss, seems to be mandatory. PMID- 11873711 TI - The case of the missing data: methods of dealing with dropouts and other research vagaries. AB - Missing data are common in most studies, especially when subjects are followed over time. This can jeopardize the validity of a study because of reduced power to detect differences, and especially because subjects who are lost to follow-up rarely represent the group as a whole. There are several approaches to handling missing data, but some may result in biased estimates of the treatment effect, and others may overestimate the significance of the statistical tests. When cross sectional data (for example, demographic and background information and a single outcome measurement time) are missing, replacement with the group mean leads to an underestimate of the standard deviation (SD) and inflation of the Type I error rate. Using regression estimates, especially with error built into the imputed value, lessens but does not eliminate this problem. Multiple imputation preserves the estimates of both the mean and the SD, even when a significant proportion of the data are missing. With longitudinal studies, the last observation carried forward (LOCF) approach preserves the sample size, but may make unwarranted assumptions about the missing data, resulting in either underestimating or overestimating the treatment effects. Growth curve analysis makes maximal use of the existing data and makes fewer assumptions. PMID- 11873712 TI - Managing schizophrenia during the stable phase: is there consensus among practice guidelines? AB - METHOD: We performed a literature search using Medline and Psycinfo databases and Google Internet search engine. RESULTS: We identified 6 clinical practice guidelines (CPGs). All stress the need for antipsychotic therapy and psychosocial interventions. Differences lie in types of antipsychotics recommended, duration of antipsychotic trial, management of extrapyramidal symptoms, and types of psychosocial interventions. Areas poorly addressed by all guidelines include definition of the stable phase of schizophrenia, management of adverse effects with atypical agents, management of clozapine nonresponders, and management of personality issues. CONCLUSION: Published CPGs are helpful in the management of the stable phase of schizophrenia, although no single CPG series appears to address all treatment needs faced by practising clinicians. PMID- 11873713 TI - Validation of French- and English-Canadian versions of the Social Cue Recognition Test. AB - OBJECTIVE: Our primary objective was to create and validate the Social Cue Recognition Test-C (SCRT-C), a Canadian test comparable with the original SCRT. METHOD: We administered the SCRT-C and the original SCRT to 111 normal undergraduate students. RESULTS: In our sample, the reliability and validity of the SCRT-C were moderately high and similar to those found with Corrigan's SCRT. The results also suggest that the English and French versions of the Canadian SCRT are equivalent. CONCLUSIONS: The SCRT-C is an appropriate instrument for assessing social cue recognition in emotional contexts. PMID- 11873714 TI - Testing for antipsychotic-induced diabetes mellitus. PMID- 11873715 TI - Quetiapine blood level variability. PMID- 11873716 TI - Atypical antipsychotic drugs and glucose dysregulation. PMID- 11873717 TI - Intravenous valproate treatment: some observations. PMID- 11873718 TI - Principal-component analysis of the Positive and Negative Syndrome Scale in patients with schizophrenia: does a 5-factor model apply to published data? PMID- 11873719 TI - Current world literature. Cerebrovascular disease. PMID- 11873720 TI - Current world literature. Neuro-ophthalmology and neuro-otology. PMID- 11873721 TI - Journals in gerontology have achieved scientific credibility but are stalled at where to go next, when changing times call for more than--or something different from--doctoral student research articles. PMID- 11873722 TI - Bankruptcy update: a new name for Vencor, continued progress. PMID- 11873723 TI - IHS' reorganization plan taking shape. PMID- 11873724 TI - [Breakthrough in thrombosis prevention?]. PMID- 11873725 TI - [Anti-smoking campaigns and their contribution to health promotion]. PMID- 11873726 TI - The saddle bridge. PMID- 11873727 TI - CRST syndrome (calcinosis, Raynaud's disease, sclerodactyly, and telangiectasias). PMID- 11873728 TI - Pierre Fidele Bretonneau 1778-1862. A pioneer in understanding infectious diseases. PMID- 11873729 TI - Living and working with pain. PMID- 11873730 TI - Re: A descriptive study of 100 patients undergoing palliative nerve blocks for chronic intractable headache and neckache. Pain Res Manage 2000;5:243-248. PMID- 11873731 TI - High dosing methadone and a possible relationship to serious cardia arrhythmias. PMID- 11873733 TI - [The physician's moral choices conditioned by the advances in contemporary medicine]. PMID- 11873732 TI - [Aleksander August Kremer (1813-1880)]. PMID- 11873734 TI - [Opinion of the Warsaw and Cracow medical circle on the situation of physicians and medicine in the modern world]. PMID- 11873735 TI - [Cracow Medical Society and its connection with the Lwow Medical Society]. PMID- 11873736 TI - [Associate professor Oscar Liszka, pioneer of surgery in Poland in the 40th anniversary of the first operation]. AB - Oscar Liszka was the first in Poland who performed operations on human brain with the use of stereotactic apparatus. It took place in 1961 in the department of Neurosurgery in Cracow. He introduced Guiot-Gultingham method, which was modified in 1964 by Wicentowicz, Goscinski and Kunicki. Since 1964 stereotactic neurosurgical operations were carried out in the Department of Experimental and Clinical Medicine, Polish Academy of Sciences in Warsaw by E. Mempel. He introduced the original Talairach method and Coopers in Parkinson syndrome. Several years later, J. Bidzinski used the apparatus of Leksell at the Department of Neurosurgery, Medical Academy in Warsaw, stereotactic procedures were also performed by Mert and Bromowicz in Lodz, by Trojanowski and Turowski in Lublin, by Slosarek in Szczecin, by Sloniewski in Gdansk and by Harat in Bydgoszcz, as well Koszewski and Zabek in Warsaw. The treatments aimed at compensating for the profound loss of dopamine in the substantia nigra substantially limited the number of stereotactic operations from 1970 to 1980. Recent advancements in Computerized Axial Tomography and Magnetic Resonance imaging as well as in the histological techniques have led to new opportunities for a small stereo-static biopsy for histological diagnosis of profound brain tumors. PMID- 11873737 TI - [Cracovian Program for Secondary Prevention of Ischaemic Heart Disease. The quality of care in the field of secondary prevention of ischaemic heart disease depends on where patients are treated in the postdischarge period]. AB - Even though the majority of actions undertaken within the secondary prevention ischaemic heart disease should be initiated while the patient is still hospitalized, the maximal benefit (measured as decreased cardiovascular risk) achieved depends mostly on the continuation and modifications of those actions in the post-discharge period. There is not much known about the quality of medical care provided for patients after hospitalization due to ischaemic heart disease. The aim of the study was to assess the quality of post-discharge care in the field of secondary prevention of ischaemic heart disease in patients treated in hospital outpatients (HO), private practice (PP), and by general practitioners (GP). METHODS: Consecutive patients (age>70 years; residing in the Cracow province) were identified according to the following clinical diagnoses or procedures: acute myocardial infarction, unstable angina, coronary artery bypass grafting and percutaneous transluminal coronary angioplasty. Risk factors and medication used were assessed 6-18 months after discharge. RESULTS: There was no significant differences in smoking, high blood pressure (>140/90 mmHg), and high fasting glucose (>6.0 mmol/l) between the study groups. High total cholesterol was found in 57.5%, 71.2% (p<0.05 vs HO) and 76.2% (p<0.05 vs HO) patients from HO, GP and PP group, respectively. Antiplatelet drugs were used in 83.5%,72.9% (p<0.05vs HO) and 67.4% (p<0.05vsHO), beta-blockers in 65.4%, 54.2% (p<0.05 29) (OR 1.46, 95%CI 1.12-1.90). A tendency towards an increased risk was found for involuntary childlessness, spontaneous abortion, thyroid drugs, and nonsteroid anti-inflammatory drugs. CONCLUSIONS: Some known risk factors for cardiac defects (eg, maternal diabetes mellitus and the use of antiepileptics) could be identified. Other postulated risk factors could not be verified, for example, paternal age and parental occupation. The use of medicinal drugs seems not to be a major factor in the etiology of cardiac defects. It is possible, however, that there is an association with the use of nonsteroid anti inflammatory drugs or drugs for thyroid disease. The relationship between a high BMI and cardiovascular malformation observed in this study may be explained by impaired maternal glucose tolerance. PMID- 11873777 TI - Medical-research official cites ethics woes. PMID- 11873778 TI - Angiotensin-receptor blockers, type 2 diabetes, and renoprotection. PMID- 11873779 TI - Angiotensin-receptor blockers, type 2 diabetes, and renoprotection. PMID- 11873780 TI - Tourette's syndrome. PMID- 11873781 TI - Adverse events after imatinib mesylate therapy. PMID- 11873782 TI - Introduction. PMID- 11873783 TI - The University of Leiden: an eclectic institution. AB - Leiden University was founded in 1575, not only in the midst of great political turmoil, but also in a time that experimented intensely with new forms of higher education. In due course Leiden was to choose an eclectic attitude, remaining loyal on the one hand to late medieval, scholastic traditions, but on the other hand emancipating the arts faculty in agreement with humanist ideas. The thesis this article wants to examine is that the curriculum of Leiden University during the first 75 years of its existence was characterised by a high level of pre university, Latin schooling, and, linked up with this, a differentiation and specialisation of the arts faculty. These developments, however, had social rather than scientific goals. The arts courses did not prepare the way for a well defined profession, but served as an initiation into a cultural elite PMID- 11873784 TI - "Professionalization" and "confessionalization": the place of physics, philosophy, and arts instruction at Central European academic institutions during the Reformation era. AB - During the sixteenth and early seventeenth centuries, physics was regularly taught as part of instruction in philosophy and the arts at Central European schools and universities. However, physics did not have a special or privileged status within that instruction. Three general indicators of this lack of special status are suggested in this article. First, teachers of physics usually were paid less than teachers of most other university-level subject-matters. Second, very few Central European academics during this period appear to have made a career out of teaching physics. And third, Reformation Era schools and universities in Central Europe emphasized language instruction; such instruction not only was instrumental in promoting the confessional--i.e., Calvinist, Lutheran, and Roman Catholic--agendas of those same schools and universities, but also helped to prepare students for service in nascent but growing state governments. PMID- 11873785 TI - Britain grants embryo cloning patent. PMID- 11873786 TI - Medical journal apologizes for violating own standards. PMID- 11873787 TI - Panel seeks better monitoring of experiments using people. PMID- 11873788 TI - Death of infant is linked to hospital study of a drug: consent form had incorrect information. PMID- 11873789 TI - Experts re-examine Dr. Reiter, his syndrome and his Nazi past. PMID- 11873790 TI - Cancer researcher in South Africa who falsified data is fired. PMID- 11873791 TI - F.D.A. says researcher failed to report a second death linked to gene therapy. PMID- 11873792 TI - Companies to cut cost of AIDS drugs for poor nations. PMID- 11873793 TI - Sharing of profits is debated as the value of tissue rises. PMID- 11873794 TI - Drug companies and third world: a case study in neglect. Medicine merchants: a special report. PMID- 11873795 TI - Studies find research on women lacking. PMID- 11873796 TI - The Dutch seek to legalize long-tolerated euthanasia. PMID- 11873797 TI - Agencies urge use of affordable drug for H.I.V. in Africa. PMID- 11873798 TI - Boom in gene testing raises questions on sharing results. PMID- 11873799 TI - Company seeking donors of DNA for a 'gene trust'. PMID- 11873800 TI - '85 murder case tests limits of patient confidentiality: judge considers letting psychiatrist testify. PMID- 11873801 TI - Patient confidentiality at issue in 1985 murder case. PMID- 11873802 TI - Should a fetus's well-being override a mother's rights? PMID- 11873803 TI - Efforts to test drugs on children hasten drive for research guidelines. PMID- 11873804 TI - Judge bars testimony in murder case, citing doctor-patient privilege. PMID- 11873806 TI - New embryonic stem-cell research guidelines criticized. PMID- 11873805 TI - New voluntary standards are proposed for experiments on people. PMID- 11873807 TI - Ways to treat disease that do not devalue life. PMID- 11873808 TI - Respect for life in bioethical dilemmas--the case of physician-assisted suicide. PMID- 11873810 TI - Retraction. PMID- 11873809 TI - The ethics of electro-shock therapy. AB - Last September the Italian National Commission on Bioethics issued an opinion on the use of electro-shock (or electroconvulsive) therapy. It has now been made available in English translation and is reprinted here. The commission had been asked for an opinion by the leader of the Green Party group on Rome City Council. The request specifically asked whether it was now advisable to suspend the practice of this particular therapy. In producing its opinion, the commission's members seem to have consulted widely within Italy but not much outside. PMID- 11873811 TI - The war on drug--prices: states are taking up the fight to reduce prescription drug costs. PMID- 11873812 TI - Plan needed to fight terrorism. AB - Public health officials are urging state lawmakers to strengthen a public health system that has been in serious need of financial and political support for years. PMID- 11873813 TI - Potential mechanism of action of J5 vaccine in protection against severe bovine coliform mastitis. AB - Coliform mastitis is one of the most difficult diseases to treat in the modern dairy industry. Curative therapy with antibiotics remains only moderately effective and depends on the stage at which the disease is treated. The most successful strategies for combating coliform mastitis appear to be prevention by hygienic management or prophylactic immunization. The severity of clinical symptoms of coliform mastitis has been shown to be reduced by immunization with the Escherichia coli J5 vaccine. However, although the J5 vaccine has been licensed in the United States for about 10 years, the immunological basis of its mechanism of action is still unknown. Until now, protection by J5 vaccination has often been explained by a straightforward mechanism of enhanced antibody production resulting in increased opsonization of coliform bacteria and lipopolysaccharides (LPS). The possibility that J5 vaccination could decrease risk factors for coliform mastitis such as impaired blood polymorphonuclear neutrophil leukocyte (PMN) diapedesis has never been investigated. This review provides arguments to support the hypothesis that J5 vaccination may reduce the severity of coliform mastitis by inducing a condition of mammary gland hyper responsiveness, characterized by a T helper 1 (Th1) response and mediated by memory cells inside the mammary gland, finally resulting in enhanced PMN diapedesis upon an intramammary infection. PMID- 11873814 TI - Microbial protein production determined by urinary allantoin and renal urea sparing in normal and low protein fed Corriedale sheep. AB - The aim of the present study was to compare the amount of microbial N entering the duodenum and the efficiency of N utilisation for microbial protein synthesis in normal (NP, 17.4 g N/d) and low protein (LP, 7.5 g N/d) fed Corriedale sheep. Renal functional tests for urea handling studies, and determination of urinary allantoin as an indirect method to estimate the microbial protein production were used. Although the N intake was 57% lower in LP sheep, the microbial N production was not very different between both diets (NP: 3.99 +/- 0.01 vs. LP: 3.79 +/- 0.02 g/d, P < 0.05). The efficiency of the microbial protein synthesis in the rumen, expressed as grams of microbial N per kg of digestible organic matter apparently digested in the rumen, was not statistically different for both diets. The urinary elimination of urea was reduced by 84% in LP sheep, essentially due to an important decrease in both renal plasma flow (-63%) and glomerular filtration rate (-71%). These haemodynamic changes would also reduce the filtered load and the urinary elimination of allantoin, thereby leading to an underestimation of the amount of microbial protein entering in the duodenum. Since the renal urea spared by the kidneys remains in the blood, it limits the drop ofthe available urea for ruminal recycling consecutive to a low nitrogen diet. PMID- 11873815 TI - Urinary cortisol as an additional tool to assess the welfare of pregnant sows kept in two types of housing. AB - The use of urinary cortisol (UC) as an additional tool to evaluate sows welfare was assessed in two experiments. In a preliminary methodological experiment, the kinetics of cortisol excretion in urine was studied during an Adreno Cortico Trophic Hormone (ACTH) challenge test in 10 pregnant sows. In a second experiment, 96 primiparous sows of an experimental unit were assigned to two different housing systems: 48 animals were housed in individual pens (IP) and 48 animals in collective pens (CP) with 6 animals per pen. UC was measured at the beginning and at the end of pregnancy and compared with other welfare indicators such as behaviour or skin damage. In both experiments, UC was measured using a high pressure liquid chromatography assay. In experiment 1, UC was constant on the day before injection of ACTH, with no variations related to circadian rhythm. It began to rise 2 h after the injection, peaked between 2 to 5 h after then returned to the basal concentration on the day after the injection. In experiment 2, UC concentrations were not different between CP- and IP-housed sows but they were higher in sows exhibiting the less stereotypies in comparison with sows exhibiting the most stereotypies. The results of this study suggest that UC is a good indicator of acute stress, more convenient than plasma cortisol measurement since it is a non-invasive method avoiding restraint or catheterisation of sows. They also suggest that UC could also give additional information on the assessment of chronic stress and improve the evaluation of animal welfare if used in conjunction with other welfare indicators. PMID- 11873817 TI - Experimental model of Border Disease Virus infection in lambs: comparative pathogenicity of pestiviruses isolated in France and Tunisia. AB - Pestiviruses have been isolated from live sheep pox Tunisian vaccines. Vaccination with these vaccines caused outbreaks of Border Disease in Tunisia. In order to study more precisely the pathogenicity of these isolates, three groups of eight four month old lambs from a pestivirus-free flock were infected by the intratracheal route with a French strain (AV) and two Tunisian isolates (SN3G and Lot21). Clinical, hematological, immunological and virological parameters were evaluated. The three groups developed mild fever and leucopaenia by day 3 to 6 post infection (pi). The differences in the weight curves were not significant. Viruses were isolated from the peripheral blood buffy coat cells by day 4 to 9 pi. Antibodies were present on day 16 pi following infection by the French strain and on day 21 pi with the Tunisian isolates. The results demonstrated that SN3G and Lot21 are almost similar to the French strain used as the reference strain. In field conditions, they could induce economical losses in naive flocks, alone or in association with other pathogens. PMID- 11873816 TI - Reliable ELISAs showing differences between resistant and susceptible lines in hens orally inoculated with Salmonella Enteritidis. AB - Reliable ELISAs were investigated with the aim to select hen lines resistant to Salmonella Enteritidis and producing high levels of antibodies. In the first experiment, the relation between the humoral response and the bacteriological results was assessed on hens from the Y11 resistant line and the L2 susceptible line, orally inoculated with 10(8) CFU S. Enteritidis per animal. Anti lipopolysaccharide (LPS) IgG titres were higher but the liver and spleen were less contaminated in hens from the Y11 line than in hens from the L2 line (p = 0.013, 0.031 and 0.026 respectively). In the second experiment, the hens were inoculated orally with 1.7 x 10(8) CFU S. Enteritidis per animal in order to select the ELISA methods showing the more significant differences. ELISAs were based on LPS, flagella, LPS from rough (LPS-R) and smooth strains (LPS-S) and detected IgG and IgM antibodies from sera and yolks. No between-line host response variation was observed in the yolk, with LPS-S and R antigens nor with anti-LPS IgM in the sera. Otherwise, significant differences were encountered between hen lines with the ELISAs performed on the sera detecting anti-LPS IgG, anti-flagella IgG or IgM (p = 0.017, 0.017 and p < 0.001 respectively). When comparing the kinetics of the selected ELISAs, the IgG antibodies against LPS detected between-line variations as early as 1 to 4 weeks pi, whereas with IgG against flagella, the differences were only detected at 1 and 2 weeks pi and with IgM against flagella, the differences were significant at 1, 2, 4 and 8 weeks pi. In conclusion, resistant hen lines producing higher levels of antibodies than the susceptible hen lines may be selected with these ELISAs. PMID- 11873818 TI - Quantitative modification of the testicular structure in pigs fed with anabolic doses of clenbuterol. AB - Morphological and structural data of the testes of thirty male pigs were recorded in order to evaluate the effects of an anabolic clenbuterol treatment. Pigs aged 6 months were randomly allocated to one of three experimental groups. In two treated groups, the animals were fed with anabolic doses of clenbuterol (1 ppm); in the CLB group (n = 10) clenbuterol was given until they were slaughtered (treatment period = 3 months) whereas in the CLBW group (n = 10) the clenbuterol was withdrawn two weeks before slaughter (treatment period = 2.5 months); clenbuterol was not given to the pigs of the control group (n = 10). Stereological estimations of the tissular volume fraction and tubular volume density were applied to quantify the structural constituents of the testes. The results showed an increased volume fraction of the testicular interstitium especially in the Leydig cell population, as a result of the clenbuterol treatment. The increase in the nuclear volume fraction of the Leydig cells was the more persistent change in the variations recorded in both treated groups with respect to the control. A regression of the seminal epithelium was also recorded in the treated animals. The rest of the structural parameters were closer to the normal figures in the CLBW group, suggesting a recovery of the testicular structure when clenbuterol was withdrawn. PMID- 11873819 TI - Cross-protection of Salmonella abortusovis, S. choleraesuis, S. dublin and S. gallinarum in mice induced by S. abortusovis and S. gallinarum: bacteriology and humoral immune response. AB - Cross-protection induced by primary infection with Abortusovis and Gallinarum was examined against challenge injection with these Salmonella serotypes as well as with Dublin and Choleraesuis, the other virulent serotypes. Abortusovis induced efficient protection against the other Salmonella. Gallinarum was ineffective against Choleraesuis. Even with low multiplication in mice, the Gallinarum J91 strain induced a weak but significant protection against Dublin (same O group serotype). The antibodies in the blood of mice were tested with ELISA specific for the Salmonella antigens used to prime or to challenge animals. The Gallinarum J91 strain was detected to be more antigenic in ELISA than the other Salmonella antigens. It is difficult to conclude on a correlation between IgM or IgG antibodies and induction of protection, because of the variability in immune response according to the different serotype used. Nevertheless, the negative linkage between a number of bacteria in the spleen of mice challenged with Gallinarum and Dublin, and the level of IgM and IgG antibodies specific for the challenging serotype, showed that humoral immune response could be one element of cross-protection, mainly by the immune response against the same O serotype. PMID- 11873821 TI - Spontaneous mycotoxic nephropathy in Bulgarian chickens with unclarified mycotoxin aetiology. AB - Histopathological, biochemical and toxicological investigations of tissues and blood of normally slaughtered chickens exhibiting different frequencies (1-2%, 40 50% and above 80%) of nephropathy changes (congested or pale and enlarged kidneys) at the slaughtering meat inspection were carried out to elucidate the aetiology of nephropathies of chickens encountered in Bulgaria. A close relationship was observed between the frequency of this nephropathy and the rate of nephrotoxic mycotoxin ochratoxin A in muscles, kidneys and livers of chickens, but the levels of ochratoxin A in corresponding feed samples (0.1-0.3 ppm) were significantly lower than the levels (2-4 ppm) required to reproduce such nephropathy. Clinicomorphological changes such as nervous symptoms, vascular and oedematous changes in various internal organs and the brain, and subcutaneous or liver and kidney haemorrhages in addition to known degenerative changes in the kidneys, liver and lymphoid organs differed from the classical description of the nephropathy made in Scandinavia. The conclusion is that the Bulgarian chicken nephropathy may have a multitoxic aetiology because it cannot be explained by the concentration of ochratoxin A alone. PMID- 11873820 TI - Effect of 1-24ACTH administration on sheep blood granulocyte functions. AB - The objective of the present study was to determine the efficiency of blood neutrophils (PMN) taken from sheep during acute stress. Ten healthy Charolle sheep were sampled before treatment (T0) and 1 (T1), 2 (T2), 24 (T24) and 48 (T48) hours after 1-24ACTH administration. Ten sheep serving as the controls were sampled at the same time intervals, using saline solution instead of 1-24ACTH. At each time sampling, rectal temperature, heart rate, cortisol, glucose, non esterified fatty acids (NEFA), total and differential leukocyte counts were evaluated. PMN were isolated after centrifugation of whole blood and hypotonic lysis of RBC. Chemotaxis was evaluated on a modified Boyden chamber using a nitrate cellulose filter and both Zymosan activated serum (ZAS) and interleukin-8 (IL-8) as chemoattractants. Phagocytosis was measured using both non-opsonized latex beads and fluoresceinated yeasts opsonized with homologous serum. Superoxide (O(-)2) production was evaluated by measuring superoxide dismutase inhibitable reduction of ferricytochrome C, and adherence by a colorimetric assay of acid phosphatase activity of adherent cells. The administration of 1-24ACTH induced an acute stress reaction, indicated by the presence of clinical, biochemical and hematological changes. Adherence significantly increased from T0 to T2 in treated sheep. This might be responsible for the depression of non specific immunity in stressed animals. Studies using stressors other than 1-24 ACTH are needed to verify the influence of other components of the stress reaction on PMN functions. PMID- 11873822 TI - The influence of different food sources on cercarial production in Lymnaea truncatula experimentally infected with Digenea. AB - Experimental infections of Lymnaea truncatula with a digenean species (Fasciola gigantica, F. hepatica, or Paramphistomum daubneyi) were performed under laboratory conditions to study the effect of four sources of food (microalgae, romaine lettuce, wheat germs, or modified Boray diet) on cercarial production. The mean number of F. hepatica, F. gigantica, or P. daubneyi cercariae was significantly greater when snail food was microalgae or Boray diet. The lowest cost prices for 100 metacercariae of F. hepatica (9.7-10.1 euros) were noted when algae of Boray diet were used as food for snails. In contrast, the highest prices (18.0-18.5 euros) were found in lettuce-reared snails. The choice of a source of food for breeding the intermediate hosts of F. hepatica and collecting metacercariae must take into account the importance of the commercial demand for metacercariae and will depend on the strategy the producer wishes to develop. PMID- 11873823 TI - Antibody responses and morbidity following infection with infectious bronchitis virus and challenge with Escherichia coli, in lines divergently selected on antibody response. AB - We evaluated the association between antibody (Ab) production and disease resistance. A controlled-challenge protocol was developed to mimic natural infection and to yield a higher rate of mortality following Escherichia coli (EC) challenge. Chicks were first infected with infectious bronchitis virus (IBV) by injecting a high dose of vaccine (attenuated virus) into their air sacs and then were infected with pathogenic EC introduced intratracheally. The experimental population consisted of lines divergently selected for high (HH) or low (LL) Ab response to EC vaccination, an HH x LL cross (HL), and commercial broilers (CC). When chicks were vaccinated with EC vaccine, mean Ab titer 15 d post-EC challenge was threefold higher in HH than LL lines, but both lines exhibited very low mortality (approximately 2%). When chicks were not vaccinated prior to EC challenge, high mortality (8 to 20%) occurred in the slow-growing HH, LL, and HL lines, and much higher mortality (approximately 40%) occurred among the CC broilers that were 38% heavier than the HH, LL, and HL lines. Mean level of Ab to EC, 7 d after EC challenge, was about twofold higher in HH vs. LL chicks and intermediate in HL and CC chicks. Within each line, Ab levels were higher in chicks exhibiting colibacillosis than in healthy ones, suggesting that these Ab were produced as a result of ongoing infection but were too late to fully prevent morbidity and mortality. These results indicate that rapid growth rate substantially reduces broiler viability, whereas Ab levels produced in response to acute pathogenic challenge without prior vaccination do not contribute to disease resistance. Among the relatively slow-growing lines, mortality was about twofold higher in HH than in LL lines. This finding may confirm previous reports that without prior vaccination, high Ab response to acute challenge increases consequent mortality; alternatively, the LL line may be superior in nonspecific defense mechanisms. PMID- 11873824 TI - Effect of selection for increased body weight and increased plasma yolk precursor on developmental stability in Japanese quail. AB - Developmental stability of several Japanese quail lines was measured by bilateral asymmetry. Lines included in the study were as follows: a randombred control (R1), sublines of R1 selected for increased (HW line) and decreased (LW line) 4 wk BW, and sublines of R1 selected for increased (HP line) or decreased (LP line) total plasma phosphorus (TPP; a measure of yolk precursor in the blood) at the beginning of lay. In sublines of the HW line, the males were selected for increased 4-wk BW and the females for increased (HW-HP line) or decreased (HW-LP line) TPP. The HW, LW, HP, and LP lines were in their 41st generation of selection and the HW-HP and HW-LP lines in their 31st generation of selection. The number of birds in each line and sex subgroup was 30. The adult breeders (28 to 32 wk of age) were weighed and killed, and bilateral measurements were made of shank length, width (laterally at the dew claw), and depth (perpendicular to the dew claw), face length, and pectoralis major and p. minor weights. Data on asymmetry was expressed for the right side minus the left side as signed and absolute differences. In order to correct for the correlation between trait size and asymmetry, relative asymmetry (RA) was obtained by dividing the absolute differences between sides by the average value of both sides and multiplying by 100. All lines differed in BW at 4 wk of age with the ranking HW > HW-LP > HW-HP > LP > R1 > HP > LW. Line rankings of adult breeders were similar, except the HP and LP lines did not differ from the R1 line and the order of ranking of the HW HP and HW-LP lines was opposite that at 4 wk of age. Line differences in signed and absolute differences were significant for most bilateral traits. However, after adjustment for trait size, line differences in RA were less frequent. In general, there were few significant differences in RA for the R1 line versus the selected lines, even though inbreeding of the R1 line (19%) was less than half that of the selected lines (44 to 57%), suggesting that homozygosity did not influence developmental stability. Selection for increased or decreased BW had little influence on RA. Developmental stability tended to be higher in the lines (LP and HW-LP) selected for decreased TPP. The data indicated that bilateral asymmetry was not a good measure of developmental stability in the current study. PMID- 11873825 TI - Effects of housing type and breeding system on the reproductive capacity of the red-legged partridge (Alectoris rufa). AB - Current methods of intensive breeding of the red-legged partridge (Alectoris rufa) are based on "industrial" laying practices, including removal and artificial incubation of eggs. These procedures can alter the reproductive behavior and physiology of the birds and, therefore, may not be suitable for use in breeding programs designed to increase wild populations. This study aimed to determine the effects of intensive housing and breeding methods on the laying capacity and reproductive behavior of the red-legged partridge. In Experiment 1, 70 pairs from a commercial game farm were randomly allocated into three treatment groups and placed in differing designs of breeding cages: 8 m2 cages with solid sides (n = 30), 4 m2 cages with solid sides (n = 30), and 4 m2 cages with mesh sides (n = 10). The number of eggs laid was recorded each week. In Experiment 2, 30 pairs, placed in 30 closed 8 m2 cages, were used. Fifteen pairs were birds reared under the intensive system used on game farms, and the other 15 pairs were birds adopted by pairs of foster parents when they were less than 48 h old. The total number of eggs laid during the reproductive period was recorded. In Experiment 1, egg production was greater in pairs housed in 8 m2 cages. There were no differences in egg production between birds housed in closed or open 4 m2 cages. In Experiment 2, the rearing method did not affect egg production. In both experiments, regardless of rearing history or cage type, the numbers of eggs laid were considerably higher than published figures for wild red-legged partridges. This fact, together with the absence of incubation by 100% of the females, indicates the considerable physiological and behavioral modifications that red legged partridges have undergone due to domestication. PMID- 11873826 TI - Multiple-objective (goal) programming model for feed formulation: an example for reducing nutrient variation. AB - A multiple-objective programming (MOP) model was applied to the feed formulation process with the objectives of minimizing nutrient variance and minimizing ration cost. A MOP model was constructed for a broiler grower ration (3 to 6 wk) and formulated with a Microsoft Excel solver. Twenty-one ingredients with 17 nutrients were included in the formulation. Amino acids were based on digestible values. The following objectives were considered as soft constraints: (1) meeting the nutrient requirements; (2) meeting the ingredient restrictions; and (3) meeting nutrient ratios, including calcium to phosphorus and the relationship of amino acids to lysine (ideal amino acid ratios). Hard constraints considered were (1) a least-cost ration and (2) minimal nutrient variances for protein, methionine, and lysine. It was found that (1) the MOP model was more flexible in providing a compromise solution than a traditional feed formulation with a linear program, (2) the MOP model was able to handle several conflicting objectives simultaneously as compared to the traditional linear programming approach that could handle only one objective, and (3) the MOP model gave the best compromise solution that would satisfy multiple decision makers when trade-offs were made between the ration cost and minimum variances of protein and methionine. The MOP model is an efficient tool to assist the decision-making process through solving a series of linear/nonlinear programs and by interacting with decision-makers. PMID- 11873827 TI - Microsatellite markers linked to Salmonella enterica serovar enteritidis vaccine response in young F1 broiler-cross chicks. AB - Reduction in Salmonella enteritidis (SE) contamination is of importance for poultry production as well as for food safety. The objectives of this study were to identify potential genetic markers of antibody response to SE vaccine in young broiler chicks and then to confirm this linkage in broiler-cross offspring, as well as to explore interactions of marker alleles with dam line and sex. The initial identification of suggestive quantitative trait loci (QTL) markers for antibody response to SE vaccine was conducted by using bulked segregant analysis (BSA) with 58 microsatellite markers in a broiler breeder male line. Four unlinked microsatellites that had allele frequency differences between the high and low antibody response DNA pools were selected for subsequent analysis in a linkage study. Antibody response was measured in an F1 population (n = 379) that was derived by crossing each of four males of the broiler line with several dams from four genetically distant, highly inbred lines (Spanish, Fayoumi, and MHC congenic G-B1 and G-B2 Leghorn). These crosses enabled us to evaluate the broiler sire QTL-marker allele effects and to explore QTL interactions with the dam lines by individual genotyping. Each of the four microsatellites identified by BSA in the broiler population had a significant (P < 0.05) association with F1 population antibody response in one or more sire families. The effect of the interaction of microsatellite allele with dam line or sex on antibody response was frequently significant. Microsatellite markers linked to antibody response QTL were identified, and genetic interactions with dam line and sex were detected. PMID- 11873828 TI - Consistent production of transgenic chickens using replication-deficient retroviral vectors and high-throughput screening procedures. AB - We have developed a novel method of DNA extraction combined with a high throughput method of gene detection allowing thousands of potentially transgenic chicks to be screened quickly and reliably. By using this method and a replication-deficient retroviral vector based on avian leukosis virus (ALV), we have demonstrated germline transmission of three different transgenes. Several generations of chickens carrying intact transgenes were produced, validating the use of the ALV retroviral vectors for large-scale production of transgenic flocks. Fourth-generation chicks that were nontransgenic, hemizygous, or homozygous for the transgene were identified with the combined genetic screening methods. PMID- 11873829 TI - Fate of plasmid DNA encoding infectious bursal disease virus VP2 capsid protein gene after injection into the pectoralis muscle of the chicken. AB - The objective of this study was to determine whether recombinant plasmid DNA injected intramuscularly into chickens expressed the gene of interest in vivo and could be subsequently detected in primary and secondary lymphoid tissues with polymerase chain reaction (PCR). The VP2 capsid protein gene of the standard challenge strain (STC) of infectious bursal disease virus (IBDV) was cloned into a eukaryotic plasmid, and purified DNA was prepared. Fourteen 2-wk-old chickens were injected in the pectoral musculature with 500 microg of plasmid DNA dissolved in sterile PBS. Seven chickens were similarly injected with PBS alone. Pectoral muscle, thymus, spleen, bursa of Fabricius, and cecal tonsils were collected at 12, 24, 36, 48, 72, 96, and 168 h postinjection for detection of protein expression (in muscle) and to extract total DNA for PCR amplification of the VP2 capsid gene. Expression of VP2 was demonstrated in muscle tissue at 12 and 24 h postinjection by using an indirect immunofluorescence assay. PCR amplification with primers specific for the VP2 gene showed that the DNA was present in the thymus, spleen, and bursa of Fabricius but not in cecal tonsils. These results demonstrate that plasmid DNA injected directly into the pectoral muscle of chickens is transcribed and translated at the injection site and promptly distributed to primary and secondary lymphoid tissues. PMID- 11873830 TI - Responses of broiler chickens to cottonseed- and soybean meal-based diets at several protein levels. AB - Three experiments were conducted to evaluate the performance of broiler chicks fed diets with cottonseed meal (CSM) as the major protein source. Experiment 1 was a 3 x 2 factorial with three crude protein levels (17, 20, and 23%) by two protein sources, CSM or soybean meal (SBM). Diets were fed to male broilers (n = 840) in floor pens from 21 to 49 d of age. L-Lysine-HCl was added to keep lysine at 5.22% of protein. Protein source and level had significant (P < 0.001) effects on BW gain (BWG) and feed conversion ratio (FCR), respectively (no source by level interactions). The average BWG were 1.80, 2.00, and 2.00 kg for birds fed CSM-based diets, compared with 1.93, 2.09, and 2.21 for SBM-based diets (17, 20, and 23% protein, respectively). The average FCR were 2.56, 2.31, and 2.25 for CSM fed broilers and 2.39, 2.16, and 2.08 for SBM-fed broilers. Significant effects of protein source or level were found for percentage of chilled carcass, fillets, tenders, saddles, and fat pads. In Experiment 2, male broiler chicks (n = 336) were used to determine the lysine requirement of chicks fed a corn and CSM-based diet with 20% CSM and 6% SBM. The basal diet contained by calculation 3,200 kcal/kg of ME, 20% CP, and 0.81% lysine. Graded levels of lysine (0.81 to 1.30% in increments of 0.07%) were fed to chicks from 10 to 20 d, and BW and residual feed were measured at 20 d. The requirement, as determined by breakpoint analysis, was 1.023+/-0.01% lysine (R2 = 0.84) for BWG and 1.028+/-0.02% lysine (R2 = 0.56) for FCR. In Experiment 3, a 2 x 2 x 4 factorial arrangement of treatments involved feeding CSM or SBM to male and female broiler chicks (n = 768) from 21 to 42 d at four dietary protein levels (17, 20, 23, and 26% CP). Lysine was kept at 5.5% of the dietary protein and consisted of the calculated minimum level, established in Experiment 2, plus 7%. Protein level, but not source, had a significant effect on BWG and FCR (P < 0.01 and P < 0.001, respectively) for males. The average BWG were 1.53, 1.74, 1.78, and 1.81 kg for birds fed CSM compared to 1.46, 1.72, 1.84, and 1.82 kg for those fed SBM (17, 20, 23, and 26% CP, respectively); average FCR were 2.36, 2.14, 2.05, and 1.97 for CSM compared to 2.35,2.04, 1.87, and 1.80 for SBM. Protein source and level significantly (P < 0.05) affected feed intake. Significant effects of protein source or level were found for percentage carcass, fillet, tenders, leg quarters, and fat pads. Females had similar qualitative responses. This study showed that at slightly higher protein levels CSM could replace SBM in broiler grower diets to achieve similar performance. PMID- 11873831 TI - The effects of dietary humate supplementation on broiler growth and carcass yield. AB - The growth-promoting effect of Farmagulator DRY Humate (FH) on live performance, carcass weight, and the abdominal fat pad of broilers was studied during different feeding periods. Four hundred, 1-d-old straight-run birds were randomly distributed to 20 floor pens of an environmentally controlled house. Four dietary regimens were replicated in five pens, each containing 20 chicks, as follows: 1) birds received no added FH in the starter or grower (NAFH), 2) birds received FH from 0 to 21 d (FH0-21), 3) birds received FH from 22 to 42 d (FH22-42), 4) birds received FH from 0 to 42 d (FH0-42) in the starter and grower diets, respectively. The FH was added to the diets at 2.5 kg/per ton of feed. Starter and grower diets were formulated to meet the minimum NRC requirements for broilers and were provided as a mash feed. Body weights at 21 d were not affected by the dietary regimens. At 42 d, body weights and feed conversions of broilers were significantly affected by the dietary humate treatments. Birds fed FH22-42 weighed more than the NAFH, whereas the FH0-21 and FH0-42 were intermediate and not different from the other treatments. Feed:gain was lower for the FH22-42 and FH0-42 treatments compared to the NAFH. There was no difference in carcass yield or abdominal fat pad percentages due to feeding FH. Feeding FH during the grower period had the most beneficial effect in terms of growth and feed conversion on broiler performance. PMID- 11873832 TI - Hemodynamic responses of broiler pulmonary vasculature to intravenously infused serotonin. AB - Serotonin is a potent pulmonary vasoconstrictor actively accumulated by mammalian platelets and avian thrombocytes and released into the plasma during platelet or thrombocyte aggregation. Serotonin has been implicated in the mechanisms responsible for pulmonary hypertension in several human and animal studies. However, the role of serotonin in pulmonary hypertension syndrome (PHS, ascites) in broilers previously had not been evaluated. In the present study we evaluated the pulmonary hemodynamic responses of broilers to intravenous infusions of serotonin dissolved in 2.5% (wt/vol) mannitol solution (carrier vehicle). Carrier vehicle infusion alone had no influence on any of the hemodynamic variables. Serotonin infusion triggered rapid increases in pulmonary arterial pressure to approximately 50% above pre-infusion baseline values, accompanied by decreases in mean systemic arterial pressure and cardiac output. The peak pulmonary arterial pressure response occurred within approximately 70 s after the start of serotonin infusion and remained elevated above baseline values over the course of a 10-min infusion period. Pulmonary arterial pressure and cardiac output returned to pre infusion baseline values upon cessation of serotonin infusion, whereas mean systemic arterial pressure returned toward pre-infusion base-line values. Pulmonary hypertensive responses were associated with increased pulmonary vascular resistance (pulmonary vasoconstriction). The peak pulmonary arterial pressure attainable was inadequate to propel the normal cardiac output through the elevated pulmonary vascular resistance. Consequently, the impeded venous return to the left ventricle caused dependent reductions in stroke volume, cardiac output, and mean systemic arterial pressure. Reductions in cardiac output were associated with reductions in stroke volume but not heart rate. Any factor that reduces the pulmonary vascular capacity or increases the pulmonary vascular resistance theoretically can increase the incidence of PHS. The present study provides direct evidence that serotonin can trigger pulmonary vasoconstriction and pulmonary hypertension in broilers. PMID- 11873833 TI - Fertility of broiler breeders following categorization by the OptiBreed sperm quality index when hens are inseminated with a constant number of sperm. AB - If semen quality was known prior to insemination, sperm doses could possibly be decreased, maximizing the number of hens inseminated. The sperm quality index (SQI), an indicator of overall semen quality, is determined by the number of deflections in a light path due to sperm movement inside a capillary tube. The objectives of this study were 1) to determine the age at which the SQI becomes a static predictor of semen quality and 2) to determine if fertility of males with a higher SQI responds more favorably to insemination dose reduction than that of males with a lower SQI. Weekly from 23 to 32 wk of age, 144 Cobb males were tested for SQI. At 32 wk of age, males were placed into four groups that represented the SQI population quartiles as follows: poor, fair, good, and best. A fifth SQI group, uncategorized, was created to determine fertility of the original population by mixing equal amounts of semen from each of the four groups. Semen was collected weekly from 33 to 40 wk of age from 18 males in each of the four groups, pooled by group, and used to inseminate 30 hens per group with 50 or 100 million sperm. Eggs were collected daily, incubated, and broken out to determine fertility. Correlation coefficients between weekly SQI results and overall averages for individual males indicated that the SQI stabilized after the birds were 28 wk of age. The main effect for SQI selection revealed that the best SQI group had the highest fertility (88%), which did not differ from the good (83%) or fair group (82%) but was greater than the uncategorized group (80%). Fertilities of the top three groups and the uncategorized group were higher than the poor group (63%) (P < 0.0001, SEM 2.18). In addition, there was an interaction between SQI classification and insemination dose. Fertilities of the top three SQI groups were similar at the 50 and 100 million sperm doses. However, the poor and uncategorized SQI groups had lower fertility at the 50 million dose as compared to the 100 million dose. By categorizing males into SQI groups after 28 wk of age, insemination dose can be reduced, maximizing a male's fertilizing potential. PMID- 11873834 TI - Changes in shell membranes during the development of quail embryos. AB - The shell membrane of an avian egg acts as a bag enclosing albumen and water. At its interface with the albumen, a smooth layer of homogeneous, dense material called the limiting membrane demarcates the shell membrane. The present study aimed to investigate changes in the limiting membrane during development of quail embryos that were grown with or without being turned. Sixty-three percent of the embryos were hatched after the eggs were incubated at 39 C and in 60% humidity with automatic rotation around their long axis and with their equatorial side down, whereas the hatch rate decreased to 24% when the eggs were incubated without being turned. The width of the limiting membrane at the equatorial region of turned eggs gradually decreased from 74 nm on Days 0 to 2 of incubation to 35 nm on Day 10 and thereafter. Conversely, water permeability, measured by evaporation through the shell membrane increased from 4 to 5 nL/mm2 per min on Days 0 to 6, to 9 nL/mm2 per min on Day 12 and thereafter. In stationary eggs, the decrease in the width of the limiting membrane on the lower side of eggs was delayed until Day 8 of incubation. The water permeability of the shell membrane in this group was 51% of that of the membrane on the upper side of eggs on Day 8 of incubation. Forty to forty-four nanometers seemed to be the critical width of the limiting membrane at which high water permeation could occur. It was also shown that the albumen hinders water permeation through the membrane. These results show that (1) the limiting membrane is made thin during the development over the whole surface with egg-turning, possibly through digestion of still unknown agents, and (2) this thinning accelerates the rate of water permeation through the membrane. PMID- 11873835 TI - Antioxidant enzyme activities and mitochondrial fatty acids in pulmonary hypertension syndrome (PHS) in broilers. AB - Major objectives of this study were to assess antioxidant protection and fatty acid profile in lung mitochondria and whole liver in broilers with pulmonary hypertension syndrome [(PHS; with and without high dietary vitamin E (VE)] (Experiment 1) and in broilers that did not develop PHS but were genetically selected (S) or not selected (NS) for resistance to PHS (Experiment 2). In Experiment 1, lung mitochondrial glutathione peroxidase (GSH-Px) activity was elevated in broilers with PHS compared to controls, broilers fed high VE, and broilers fed high VE with PHS (VE-PHS), but there were no differences in GSH reductase (GSH-Rd) among groups. In liver tissue, GSH-Px was also elevated by PHS but was lower in VE and VE-PHS groups than in controls. There were no differences in liver GSH-Rd, superoxide dismutase (SOD), or gamma-glutamylcysteine synthetase (gamma-GCS) activities with the exception that gamma-GCS was higher in the VE-PHS group than in the other groups. In Experiment 2, S lung mitochondria exhibited lower GSH-Px and higher GSH-Rd compared to NS broilers. In the liver, there were no differences in GSH-Px, GSH-Rd, or gamma-GCS, but SOD was lower in S compared to the NS broilers. High VE increased the percentage of saturated fatty acids and decreased the percentage of unsaturated fatty acids in lung mitochondria in Experiment 1; there were no differences in fatty acid content between S and NS mitochondria in Experiment 2. Thus, it appears that GSH recycling enzyme activities are affected by PHS and high VE presumably in response to differences in oxidative stress and that genetic resistance to PHS is associated with an inherently better capability to metabolize oxidants in lung mitochondria. The increase in saturation of lung mitochondrial fatty acids with high dietary VE would presumably make them more resistant to oxidative stress and, thus, reduce the level of PHS-induced oxidative stress. PMID- 11873836 TI - Sex differences in central benzodiazepine receptor densities and circulating corticosterone release after acute stress in broiler chicks. AB - The purpose of the study was to determine the effect of sex on central benzodiazepine receptor (CBR) and serum corticosterone (CS) responses to an acute stressor in broiler chicks. Birds were housed in ten mixed-sex groups of eight chicks per cage. At 15 d of age, chicks were taken from a randomly selected cage and blood was immediately sampled (undisturbed controls), or they were taken from the same cage and immersed up to their necks in warm water (partial water immersion, PWI) for 15 min before blood was sampled. After blood sampling, forebrains were dissected for preparation of membranes, and bird sex was determined by gonadal inspection. Serum CS levels were determined by a competitive protein-binding assay. CBR densities were determined by radiolabeled receptor binding assay. There were no sex differences in serum CS levels or benzodiazepine receptor densities in controls. Exposure to PWI significantly increased (P < 0.01) circulating CS levels in both sexes, and this elevation was more pronounced (P < 0.01) in males than in females. Male, but not female, chicks also showed a significant stressor-induced increase (P < 0.01) in CBR densities. These findings showed sexual differences in acute, stressor-induced benzodiazepine and adrenocortical responses that suggest broiler males are more stress-susceptible than females. PMID- 11873837 TI - The effect of dietary ascorbic acid on semen traits and testis histology of male turkey breeders. AB - A 9-mo field trial was conducted to evaluate the effects of dietary L-ascorbic acid (AA) on semen traits of 144 male turkey breeders. Dietary AA treatments were initiated when birds were 30 wk of age. Semen and blood collection began at 32 wk of age. Three treatments with four pens per treatment and 12 birds per pen were fed 0, 75, and 150 mg/kg AA during the first 4 mo of their reproductive cycle. Levels of AA were doubled in the supplemented diets to 150 and 300 mg/kg during Months 5 to 9. Semen traits and blood AA were unaffected by dietary AA. When birds were 65 wk of age, testes were removed from 12 birds per treatment for histological analysis. Multinucleated giant cells (MCG), indicative of degeneration, were observed in the testes of 7 of the 12 control birds but were absent from AA-supplemented birds (P < 0.02). The antioxidant properties of AA may delay formation of these degenerative cells. In conclusion, dietary AA levels employed in the current study did not affect semen traits or testis weight but were associated with reduced formation of MGC in the testes of 65 wk-old breeder toms. PMID- 11873838 TI - Off-odor volatiles and pink color development in precooked, irradiated turkey breast during frozen storage. AB - The effect of irradiation on color, lipid oxidation, and volatile production of precooked, irradiated turkey breast during frozen storage was studied. Turkey breast muscles were precooked, aerobically or vacuum-packaged, irradiated at 0, 2.5, or 5.0 kGy using a linear accelerator (electron beam), and then frozen stored at -40 C. Lipid oxidation, volatiles, color values, gas production, and oxidation-reduction potential (ORP) of meat were determined during 3-mo storage periods. Ionizing radiation produced characteristic off-odor volatiles (dimethyldisulfide and methylthioethane) and lipid oxidation products in precooked, frozen turkey breast. The production of volatiles was accelerated by the confounding effect of high irradiation dose, aerobic packaging, and increased storage time. Volatile production and color changes in irradiated, precooked turkey breast were induced by different mechanisms. Irradiation increased pink color in precooked, vacuum-packaged turkey breast, and the pink color was stable during frozen storage. Decreased ORP and increased CO in irradiated meat indicated that denatured CO-heme pigments could be responsible for the pink color in precooked, irradiated turkey breast. Vacuum packaging was better than aerobic packaging in preventing lipid oxidation and oxidation-dependent volatile production, but pink color in precooked, irradiated turkey breast during frozen storage was maintained. PMID- 11873839 TI - Effects of raw broiler breast meat color variation on marination and cooked meat quality. AB - Three replicate trials were conducted to determine the effect of raw broiler breast meat color on marinated and cooked meat quality. In each trial, 90 fillets were collected from a commercial processing plant based on lightness (L*) values of breast meat as follows: light, L* > 53; normal, 48 < L* < 51; and, dark, L* < 46. For each fillet the color, pH, and weight were determined, and the fillets were marinated in three lots of 10 fillets from each color group (20% wt:wt of 5% salt and 2.5% sodium tripolyphosphate in a vacuum tumbler). Marination uptake was determined; the samples were held for 24 h at 2 C, and color, pH, and weight again were determined. Next, the samples were cooked, and the meat was subjected to color, pH, cooked yield, shear force, and moisture analyses. Results showed that absolute color values changed with marination and cooking, but that L* and pH differences by color group were maintained through marination and cooking. There were also significant differences in marinade absorption, cooked yield, and shear value, particularly among the extremes of light and dark meats. There was no significant difference in final cooked meat moisture. As expected, there were significant negative correlations between meat lightness and pH; however, raw muscle pH was not correlated to final product moisture or shear but was positively correlated to cooked meat yield and negatively correlated to marinade absorption. These results indicate that the pH variation associated with extreme raw breast meat color variation can affect breast meat marination and cooked meat quality. PMID- 11873840 TI - Rearrangement of receptive field topography after intracortical and peripheral stimulation: the role of plasticity in inhibitory pathways. AB - Intracortical microstimulation (ICMS) of a single site in the somatosensory cortex of rats and monkeys for 2-6 h increases the number of neurons responsive to the skin region corresponding to the ICMS-site receptive field (RF), with very little effect on the position and size of the ICMS-site RF, and the response evoked at the ICMS site by tactile stimulation. Large changes in RF topography are also observed following several weeks of repetitive stimulation of a restricted skin region during tactile frequency discrimination training in monkeys. It has been suggested that these changes in RF topography are caused by competitive learning in excitatory pathways. This paper analyses the possible role of lateral inhibitory synaptic plasticity in producing cortical plasticity after ICMS and peripheral conditioning in adult animals. The 'EXIN' (afferent excitatory and lateral inhibitory) synaptic plasticity rules are used to model RF changes after ICMS and peripheral stimulation. The EXIN model produces RF topographical changes similar to those observed experimentally. It is shown that lateral inhibitory pathway plasticity is sufficient to model RF changes and increase in position discrimination after peripheral stimulation. Several novel and testable predictions are made based on the EXIN model. PMID- 11873841 TI - Pattern storage and processing in attractor networks with short-time synaptic dynamics. AB - Neurophysiological experiments show that the strength of synaptic connections can undergo substantial changes on a short time scale. These changes depend on the history of the presynaptic input. Using mean-field techniques, we study how short time dynamics of synaptic connections influence the performance of attractor neural networks in terms of their memory capacity and capability to process external signals. For binary discrete-time as well as for firing rate continuous time neural networks, the fixed points of the network dynamics are shown to be unaffected by synaptic dynamics. However, the stability of patterns changes considerably. Synaptic depression turns out to reduce the storage capacity. On the other hand, synaptic depression is shown to be advantageous for processing of pattern sequences. The analytical results on stability, size of the basins of attraction and on the switching between patterns are complemented by numerical simulations. PMID- 11873842 TI - The role of intra-thalamic and thalamocortical circuits in action selection. AB - We previously proposed that the basal ganglia (BG) play a crucial role in action selection. Quantitative analysis and simulation of a computational model of the intrinsic BG demonstrated that its output was consistent with this proposition. Here we build on that model by embedding it into a wider circuit containing the motor thalamocortical loop and thalamic reticular nucleus (TRN). Simulation of this extended model showed that the additions gave five main results which are desirable in a selection/switching mechanism. First, low salience actions (i.e. those with low urgency) could be selected. Second, the range of salience values over which actions could be switched between was increased. Third, the contrast between the selected and non-selected actions was enhanced via improved differentiation of outputs from the BG. Fourth, transient increases in the salience of a non-selected action were prevented from interrupting the ongoing action, unless the transient was of sufficient magnitude. Finally, the selection of the ongoing action persisted when a new closely matched salience action became active. The first result was facilitated by the thalamocortical loop; the rest were dependent on the presence of the TRN. Thus, we conclude that the results are consistent with these structures having clearly defined functions in action selection. PMID- 11873843 TI - Viewpoint-dependent recognition of scenes. AB - Three experiments investigated scene recognition across viewpoint changes, involving same/different judgements on scenes consisting of three objects on a desktop. On same trials, the comparison scene appeared either from the same viewpoint as the standard scene or from a different viewpoint with the desktop rotated about one or more axes. Different trials were created either by interchanging the locations of two or three of the objects (location change condition), or by rotating either one or all three of the objects around their vertical axes (orientation change condition). Response times and errors increased as a function of the angular distance between the standard and comparison views, but this effect was bigger for rotations around the vertical axis than for those about the line of sight or horizontal axis. Furthermore, the time to detect location changes was less than that to detect orientation changes, and this difference increased with increasing angular disparity between the standard and comparison scenes. Rotation times estimated in a double-axis rotation were no longer than other rotations in depth, indicating that alignment was not necessarily simpler around a "natural" axis of rotation. These results are consistent with the hypothesis that scenes, like many objects, may be represented in a viewpoint dependent manner and recognized by aligning standard and comparison views, but that the alignment of scenes is not a holistic process. PMID- 11873844 TI - Feature sampling in categorization and recognition of objects. AB - This article presents an overview of some recent work on the time course of perceptual categorization and recognition memory. First, The EGCM-RT, which is a feature-sampling model of the time course of categorization, is described. It is shown that the model explains a wide range of categorization data. Second, an overview is given of a feature-sampling model of the time course of recognition that is derived from the EGCM-RT. This model explains results that have been interpreted in the past as evidence for dual-process models of recognition, and it provides a single-process alternative to dual-process accounts. PMID- 11873845 TI - Timing goals in bimanual coordination. AB - Phase coupling between movement trajectories has been proposed as the basic mechanism of hand coordination in the production of bimanual rhythmic movements with a 1:2 frequency ratio. Here a central temporal coupling view is proposed as an alternative. Extending previous models of two-handed synchronic and alternate hand tapping, we hypothesized that 1:2 tapping is performed under the control of a single internal timekeeper set at the frequency required for the fast hand. The fast hand is assumed to use every signal and the slow hand every other signal of the timekeeper, to produce actions coordinated in time. The model's predictions for the variance-covariance pattern of tap timing within and across hands were tested in an experiment that required tapping with both hands with 1:1 or 1:2 frequency ratio. The finger contact on the response plate was to be short or long, according to instruction. Prolonged finger contact entailed profound modifications in the movement trajectories but failed to modify the variance covariance pattern of the tap timing. This pattern proved to conform to predictions under both the short and the long contact conditions, thus supporting the central temporal coupling hypothesis. PMID- 11873846 TI - Responding under time pressure: testing two animal learning models and a model of visual categorization. AB - Two experiments are reported, which employed a Pavlovian eyelid conditioning procedure with human participants. The experiments tested the predictions of three models of the time-course of processing under time pressure. These were the extended generalized context model (Lamberts, 1998), and two variants of the Rescorla-Wagner model (Rescorla & Wagner, 1972), which were activated in cascade mode. Reinforcement schedules in the experiments were equivalent either to an AND rule or to an XOR rule. The time available for processing the conditioned stimulus and initiating a conditioned response was manipulated by varying the interval from the onset of the conditioned stimulus to the onset of the unconditioned stimulus. The results were in accord with the predictions of one of the two variants of the Rescorla-Wagner model. PMID- 11873847 TI - Verbal short-term memory as an articulatory system: evidence from an alternative paradigm. AB - In a series of experiments, the role of articulatory rehearsal in verbal [corrected] short-term memory was examined via a shadowing-plus-recall paradigm. In this paradigm, subjects shadowed a word target presented closely after an auditory memory list before they recalled the list. The phonological relationship between the shadowing target and the final item on the memory list was manipulated. Experiments 1 and 2 demonstrated that targets sounding similar to the list-final memory item generally took longer to shadow than unrelated targets. This inhibitory effect of phonological relatedness was more pronounced with tense- than lax-vowel pseudoword recall lists. The interaction between vowel tenseness and phonological relatedness was replicated in Experiment 3 using shorter lists of real words. In Experiment 4, concurrent articulation was applied during list learning to block rehearsal; consequently, neither the phonological relatedness effect nor its interaction with vowel tenseness emerged. Experiments 5 and 6 manipulated the occurrence frequencies and lexicality of the recall items, respectively, instead of vowel tenseness. Unlike vowel tenseness, these non-articulatory memory factors failed to interact with the phonological relatedness effect. Experiment 7 orthogonally manipulated the vowel tenseness and frequencies of the recall items; slowing in shadowing times due to phonological relatedness was modulated by vowel tenseness but not frequency. Taken together, these results suggest that under the present paradigm, the modifying effect of vowel tenseness on the magnitude of slowing in shadowing due to phonological relatedness is indicative of a prominent articulatory component in verbal short term retention. The shadowing-plus-recall approach avoids confounding overt recall into internal memory processing, which is an inherent problem of the traditional immediate serial recall and span tasks. PMID- 11873848 TI - The influence of only on syntactic processing of "long" relative clause sentences. AB - We report an eye movement experiment investigating the influence of the focus operator only on syntactic processing of "long" relative clause sentences. Paterson, Liversedge, and Underwood (1999) found that readers were garden pathed by "short" reduced relative clause sentences containing the focus operator only. They argued that due to thematic differences between "short" and "long" relative clause sentences, garden path effect might not occur when "long" reduced relative clause sentences are read. Eye-tracking data show that garden path effects found during initial processing of the disambiguating verb of "long" reduced sentences without only were absent or delayed in the case of counterparts with only. We discuss our results in terms of current theories of sentence processing. PMID- 11873849 TI - Modality-specific auditory and visual temporal processing deficits. AB - We studied the attentional blink (AB) and the repetition blindness (RB) effects using an audiovisual presentation procedure designed to overcome several potential methodological confounds in previous cross-modal research. In Experiment 1, two target digits were embedded amongst letter distractors in two concurrent streams (one visual and the other auditory) presented from the same spatial location. Targets appeared in either modality unpredictably at different temporal lags, and the participants' task was to recall the digits at the end of the trial. We evaluated both AB and RB for pairs of targets presented in either the same or different modalities. Under these conditions both AB and RB were observed in vision, AB but not RB was observed in audition, and there was no evidence o PMID- 11873850 TI - Goal specificity effects on hypothesis testing in problem solving. AB - Previous research has found that having a nonspecific goal (NSG) leads to better problem solving and transfer than having a specific goal (SG). To distinguish between the various explanations of this effect requires direct evidence showing how a NSG affects a participant's behaviour. Therefore we collected verbal protocols from participants learning to control a linear system consisting of 3 outputs by manipulating 3 inputs. This system was simpler than the one we had used previously, so in Exp. 1 we generalized our earlier goal specificity findings to this system. In Exp. 2 protocol analysis confirmed our prediction (based on dual-space theories of problem solving) that NSG participants focused on hypothesis testing whereas SG participants focused on the goal. However, this difference only emerged over time. We also replicated the goal specificity effect on performance and showed that giving participants a hypothesis to test improved performance. PMID- 11873851 TI - Verbal structure of numerals and digits handwriting: new evidence from kinematics. AB - Two experiments used a digitizing tablet to analyse the temporal, spatial, and kinematic characteristics of handwritten production of arabic numbers. They addressed a specific issue of the numerical domain: Does the lexical and syntactic structure of verbal numerals influence the production of arabic numerals (Experiments 1 and 2), even after enforced semantic processing in a comparison task (Experiment 2)? Subjects had to write multi-digit arabic numerals (e.g., 1200) presented in two different verbal structures: a multiplicative one (e.g., teen-hundred, douze cents (twelve hundred)) or an additive one (e.g., thousand-unit-hundred, mille deux cents (one thousand two hundred)). Results show differences in the inter-digit jumps that reflect the influence of the structure of verbal numerals, even after the semantic task. This finding is discussed with regard to different models of number transcoding (McCloskey, Caramazza, & Basili, 1985; Power & Dal Martello, 1990, 1997). PMID- 11873852 TI - Global precedence and response activation: evidence from LRPs. AB - Lateralized readiness potentials (LRPs) were measured in left/right/no-go tasks using compound global/local stimuli. In Experiment 1, participants responded to local target shapes and ignored global ones. RTs were affected by the congruence of the global shape with the local one, and LRPs indicated that irrelevant global shapes activated the responses with which they were associated. In Experiment 2, participants responded to conjunctions of target shapes at both levels, withholding the response if a target appeared at only one level. Global shapes activated responses in no-go trials, but local shapes did not. The results are consistent with partial-output models in which preliminary information about global shape can partially activate responses that are inconsistent with the local shape. They also demonstrate that part of the global advantage arises early, before response activation begins and probably before recognition of the local shape. PMID- 11873853 TI - How not to revisit Highway 61: negative repetition effects in a post-cue naming task. AB - Repetition effects were studied in a post-cue naming task, in which participants were cued to name one of two stimuli following their presentation. When pairs of pictures were repeated in a second block, former distractors (not named in Block 1) were named faster than former targets (named in Block 1). This negative repetition effect was not found when two words rather than two pictures were used or when a semantic categorization task was used with two pictures. From this we conclude that the effect reflects a process of mapping from a semantic representation to a name. Negative repetition was not found with a simultaneous selection cue, suggesting that it arose only when there was competition for name selection. It was also dependent on memory for previous acts of semantic naming. We propose that negative repetition reflects a form of speech monitoring that is applied when there is competition in the process of mapping from semantic to name representations. PMID- 11873854 TI - Conditionals and directionality: on the meaning of if vs. only if. AB - The aim of this study was to test the predictions of the current theories of reasoning about the comprehension of conditional statements. We used two types of conditional statement that are logically equivalent: if p then q and p only if q. The model theory of reasoning considers that these conditional forms differ in their initial meaning, because the negative contingency is considered only in the p only if q form. Mental-rule theories maintain that the interpretation of p only if q depends on a rephrasing of the statement as: if not q then not p. Alternatively, a directional bias may explain the differences between if p then q and p only if q. We report three experiments that demonstrate the existence of a directional bias in the comprehension of the conditionals. The results were not predicted by either the mental-rules theories or the model theory; they could, however, be easily assimilated by the model theory. PMID- 11873855 TI - Segmentation and selection contribute to local processing in hierarchical analysis. AB - The present study examined the role of segmentation and selection processes when we respond to local elements in hierarchical stimuli. The ease of segmentation and selection of an individual local element from hierarchical patterns was manipulated by making one local element substantially distinct from the others in colour. Experiment 1 showed that, when attention was spread across the global and local levels in a divided attention task, the introduction of the local red element speeded responses to local targets but slowed responses when targets appeared at the global level. Experiment 2 used a selective attention task in which subjects responded only to the local or the global shapes across a block of trials. Under these circumstances, the local red element reduced global-to-local interference in addition to speeding local responses. The results suggest that the efficiency with which local elements are segmented and selected affects responses to local aspects of hierarchical patterns; furthermore, the effect of local pop-out on global processing is contingent on top-down attentional control settings. PMID- 11873856 TI - Influences of different combinations of conceptual, perceptual, and structural similarity on stimulus-response compatibility. AB - This study evaluated the hypothesis that an increase in set-level stimulus response compatibility produces facilitation for congruent mappings and interference for incongruent mappings. The degree of set-level compatibility was manipulated by varying combinations of conceptual, perceptual, and structural similarity. Experiment 1 varied perceptual similarity, by combining two stimulus codes (spatial, verbal) with two response modalities (manual, vocal) for orthogonal spatial dimensions, which have structural similarity. The element level mapping effect did not vary as a function of the code-modality relation, in contrast to findings obtained with parallel spatial dimensions, which also have conceptual similarity. Experiment 2 manipulated combinations of conceptual and perceptual similarity by combining vertical and horizontal stimulus and response orientations, using verbal or spatial stimuli and vocal responses. The element level mapping effect was larger for parallel than orthogonal orientations, with congruent mappings showing facilitation and incongruent mappings showing interference. The largest effect was facilitation for parallel orientations with the verbal-vocal set, consistent with the view that perceptual similarity contributes to performance primarily when responding with the identity of the stimulus. Our results indicate that conceptual similarity, but not perceptual similarity, produces the facilitation/interference pattern suggestive of automatic activation of the corresponding response regardless of mapping. PMID- 11873857 TI - Plausibility and subcategorization preference in children's processing of temporarily ambiguous sentences: evidence from self-paced reading. AB - Three self-paced reading experiments investigated children's processing of temporarily ambiguous sentences. Across the three experiments, subcategorization preference of a verb in a subordinate clause and the semantic plausibility of the misanalysis were manipulated. Reading times in the temporarily ambiguous region and following syntactic disambiguation indicated that children in the age range tested (8 years, 11 months to 12 years, 11 months) routinely misanalyse sentences of the type tested, and their tendency to misanalyse the sentences does not depend on the subcategorization preferences of the initial verb. Additional correlational analyses suggested that subcategory information did affect the degree of difficulty that readers experienced processing the critical noun and matrix verb. PMID- 11873858 TI - Perceiving a strong causal relation in a weak contingency: further investigation of the evidential evaluation model of causal judgement. AB - Contingency information is information about the occurrence or nonoccurrence of a certain effect in the presence or absence of a candidate cause. An objective measure of contingency is the deltaP rule, which involves subtracting the probability of occurrence of an effect when a causal candidate is absent from the probability of occurrence of the effect when the candidate is present. Causal judgements conform closely to deltaP but deviate from it under certain circumstances. Three experiments show that such deviations can be predicted by a model of causal judgement that has two components: a rule of evidence, that causal judgement is a function of the proportion of relevant instances that are judged to be confirmatory for the causal candidate, and a tendency for information about instances in which the candidate is present to have greater effect on judgement than instances in which the candidate is absent. Two experiments demonstrate how this model accounts for some recently published findings. A third experiment shows that it is possible to use the model to predict the occurrence of high causal judgements when the objective contingency is close to zero. PMID- 11873860 TI - The regulation and physiological roles of the guanylyl cyclase receptors. PMID- 11873859 TI - Fibroblast growth factor (FGF)-23 and hypophosphatemic rickets/osteomalacia. PMID- 11873861 TI - Vascular endothelial growth factors and thyroid disorders. AB - Using thyroid follicles in suspension culture, in which thyroid function is maintained for a couple of weeks, while preserving Wolff-Chaikoff effect and responding to physiological concentrations of TSH (0.1 microU/ml), we have partly elucidated the pathophysiology of increased blood flow in Graves' thyroid gland. Furthermore, we are investigating the mechanism by which iodide inhibits thyroid blood flow. Since administration of a large dose of iodide prior to subtotal thyroidectomy decreases thyroid blood flow and reduces operative morbidity and mortality in patients with Graves' disease, it is important to elucidate the mechanism by which thyroid blood flow is regulated. Furthermore, this research field will be clinically important to develop new strategies for the treatment of hypervascular and aggressive thyroid carcinoma, particularly anaplastic thyroid carcinoma. A recent report that anti-VEGF antibody reduced cancer growth in nude mice transplanted with human thyroid carcinoma is an indication this field holds for future studies. PMID- 11873862 TI - Norepinephrine inhibits glucose-stimulated, Ca2+-independent insulin release independently from its action on adenylyl cyclase. AB - Inhibition of insulin release by norepinephrine has been attributed to activation of ATP-sensitive K+ channels, inactivation of voltage-dependent Ca2+ channels, and inhibition of adenylyl cyclase. However, direct inhibitory action of norepinephrine at a distal site of stimulus-secretion coupling has also been suggested. To obtain more direct evidence for norepinephrine inhibition of insulin release at a distal site, we performed experiments in intact, non permeabilized beta cells. In rat pancreatic islets, a combination of glucose, phorbol ester and forskolin under stringent Ca2+-free conditions was used as a trigger of insulin exocytosis at a distal site. Norepinephrine inhibited this Ca2+-independent insulin release in a concentration-dependent manner, with an IC50 of 50 nM. The inhibition was complete, reversible, and pertussis toxin sensitive, and not associated with any reduction of cAMP content in the islet cells. In conclusion, norepinephrine strongly, yet reversibly, inhibits insulin release in intact beta cells at a late step of exocytosis, through pertussis toxin-sensitive, G protein-mediated mechanism(s). PMID- 11873863 TI - Neonatal exposure to genistein reduces expression of estrogen receptor alpha and androgen receptor in testes of adult mice. AB - We investigated the long-term estrogenic influence of genistein on the male reproductive system in mice. Newborn ICR male mice were treated with genistein (10, 100, or 1,000 microg/mouse) for neonatal 5 days. As positive control, administration of diethylstilbestrol (0.5-50 microg/mouse) was carried out. In mice exposed to genistein,we examined weight of testes, sperm counts, sperm motility, and mRNA expression levels of estrogen receptor a (ERalpha) and androgen receptor (AR) at 4, 8 or 12 weeks after birth. Moreover, at 12 weeks of age, we evaluated protein level of ERalpha. In our conventional reproductive toxicological study (weight of testes, sperm counts and sperm motility), neonatal transient exposure to genistein did not show adverse effects on the male reproductive system in 4, 8 or 12 week old mice. However, in mice treated with genistein mRNA expression levels of ERa and AR were reduced at 8 weeks. This reduction was recovered at 12 weeks in mice treated with a lower dose (10 microg) of genistein but not in those with higher doses (100 microg and 1,000 microg). In addition, ERa protein levels tended to decrease in 12 weeks of adulthood. Our results exhibited that the disruption of gene expression continued for long term such as 3 months after administration of genistein, even if no effect was found at conventional reproductive-toxicological level. We have shown that neonatal administration of weak estrogenic compound (genistein) affects male reproductive organs at molecular levels in adulthood. PMID- 11873864 TI - Rational, effective metyrapone treatment of ACTH-independent bilateral macronodular adrenocortical hyperplasia (AIMAH). AB - Standard therapy for ACTH-independent bilateral macronodular adrenocortical hyperplasia (AIMAH), a rare form of Cushing's syndrome, is bilateral adrenalectomy. Patients with AIMAH are usually elderly, with a variety of complications, and at risk for surgery. Postoperatively, they must receive lifelong corticosteroids and spend the remainder of their lives avoiding adrenal crisis. Therapy using metyrapone, a potent inhibitor of steroidogenesis, provides the advantages of avoiding the surgery. Its effectiveness is further anticipated because adrenal steroidogenic enzymes are reportedly weak in AIMAH. Treatment with metyrapone thus appears a good therapy for AIMAH, but its effectiveness has not, to our knowledge, been studied. We treated a 59-year-old man with AIMAH with metyrapone. At a low dose of metyrapone (500 to 750 mg/day), his plasma cortisol levels decreased to the normal range, and hypertension and diabetes mellitus were ameliorated. Therapy using metyrapone thus appears effective in treating AIMAH, and can be recommended for high risk AIMAH patients as an alternative therapy. PMID- 11873865 TI - Production of 8-OHdG and cytochrome c by cultured human mononuclear cells in patients with autoimmune thyroid disease. AB - Since oxidative stress is related to autoimmune thyroid disease, we studied the production of 8-hydroxy-2'-deoxyguanosine (8-OHdG) and cytochrome c by culture of mononuclear cells from patients with Graves' disease and Hashimoto's thyroiditis. In patients with untreated Graves' disease, 8-OHdG and cytochrome c levels in culture supernatant of mononuclear cells were significantly higher than those of healthy control subjects, while the cytochrome c levels were significantly higher in patients with untreated Graves' disease and Hashimoto's thyroiditis than those of control subjects. Significant correlations between 8-OHdG and FT4, and cytochrome c were found. These results indicated that thyroid function has a potent influence on oxidative stress. PMID- 11873866 TI - A natural history of adrenocorticotropin-independent bilateral adrenal macronodular hyperplasia (AIMAH) from preclinical to clinically overt Cushing's syndrome. AB - A 49-year-old man was referred to our hospital for the treatment of gallstones in 1993. Bilateral adrenal nodular masses were detected incidentally by abdominal computed tomography. He had no clinical signs of Cushing's syndrome such as central obesity, striae of skin and diabetes mellitus. We performed cholecystectomy and partial adrenalectomy of right adrenal gland as a biopsy, and diagnosed him as preclinical Cushing's syndrome due to adrenocorticotropin independent bilateral adrenal macronodular hyperplasia (AIMAH) based on endocrinological and histological examinations. We followed him up for 7 years. During the observation period, the sizes of both adrenal glands increased gradually, and finally serum cortisol level increased beyond normal range, and he showed a Cushingoid appearance such as moon face and central obesity. His skin became atrophic and very fragile, and the bone mineral density of his lumbar spine was extremely low. Serum cortisol level was elevated, and plasma ACTH level was always suppressed. Urinary excretion of 17-hydroxycorticosteroid and free cortisol were increased. Diurnal rhythm of cortisol and ACTH was completely lost and high dose (8 mg/day) dexamethasone did not suppress urinary 17 hydroxycorticosteroid excretion. He became clinically overt Cushing's syndrome. We recommended total adrenalectomy, but he refused it. It is important to know the natural history of preclinical Cushing's syndrome due to AIMAH when choosing an adequate treatment. PMID- 11873867 TI - Relationships among serum testosterone levels, body fat and muscle mass distribution in women with polycystic ovary syndrome. AB - We investigated the relationships among serum testosterone levels, body fat and muscle mass distribution in women with polycystic ovary syndrome (PCOS). Subjects were 67 women with PCOS (mean age +/- standard deviation, 28.8 +/- 6.6 years). Baseline characteristics included age and height. Trunk-leg fat ratio and trunk leg muscle ratio were assessed with dual-energy x-ray absorptiometry. Serum testosterone and dehydroepiandrosterone sulfate levels were measured with radioimmunoassays. Relationships among serum testosterone levels, body fat and muscle mass distribution were investigated using Pearson and partial correlation tests. Serum testosterone levels were positively correlated with trunk-leg fat ratio (r = 0.398, P < 0.01), but were inversely correlated with trunk-leg muscle ratio (r = -0.332, P < 0.05). Trunk-leg muscle ratio was inversely correlated with trunk-leg fat ratio (r = -0.360, P < 0.01). Serum testosterone levels were still correlated with trunk-leg fat ratio (r = 0.500, P < 0.001) and trunk-leg muscle ratio (r = -0.286, P < 0.05), after adjusting for age and height. Trunk leg fat ratio was still correlated with trunk-leg muscle ratio, after adjusting for age, height, and serum testosterone levels. Based on these results, we concluded that higher serum testosterone levels may contribute to the upper body fat distribution and peripheral muscle mass distribution. In addition, peripheral muscle mass distribution may also contribute to the upper body fat distribution. PMID- 11873868 TI - Management of massive retroperitoneal hemorrhage from an adrenal tumor. AB - Spontaneous massive retroperitoneal hemorrhage from an adrenal gland is a rare event. A thoughtful and meticulous approach to such a patient, with appropriate diagnostic studies, ICU and surgical care are essential for patient survival. In patients with active bleeding, angiographic embolization is a valuable adjunct to achieve hemostasis, to allow for further work-up of the adrenal tumor, and an improved subsequent oncologic resection. Hemodynamically unstable patients, however, may require supportive transfusions in the intensive care unit, potential embolization if deemed feasible, or urgent surgical exploration. If possible, however, the acute surgical removal of an adrenal tumor within a large retroperitoneal hematoma should be avoided, as under such conditions a proper oncologic resection may not be possible. The possibility of a pheochromocytoma must always be entertained. Early recognition and treatment of patients with presumed adrenal insufficiency may decrease patient morbidity and mortality. PMID- 11873869 TI - Thallium-201 scintigraphy was useful in diagnosing ectopic ACTH syndrome due to bronchial carcinoid. AB - Abstract. Initial investigations of a 70-year-old woman with clinical Cushing's syndrome, including overnight dexamethasone suppression test, CRH test, and pituitary MRI, suggested the presence of ectopic ACTH production. Thoracic computed tomography (CT) scan revealed a mass measuring 7 mm in the right lung, but it was thought to be an incidental opacity, leaving the source of ectopic ACTH undetermined for several years. During this period, although the size of the lung opacity did not change remarkably, serum cortisol levels became elevated to 43 microg/dl, and the patient's symptoms worsened. Tl-201 SPECT demonstrated intense accumulation in the right lung. The mass was surgically resected using thoracoscopy to investigate it as the focus of ACTH production. Histological and immunohistochemical examination confirmed that the area of intense Tl-201 uptake was an ACTH-producing bronchial carcinoid. Plasma ACTH and cortisol levels decreased immediately after the surgery. In conclusion, this case demonstrated Tl 201 scintigraphy as a useful tool in identifying the location of an ACTH producing bronchial carcinoid. PMID- 11873870 TI - Blocking type anti-tSH receptor antibodies detected by radioreceptor assay in Graves' disease. AB - We previously developed a radioreceptor assay which is presumably specific for detection of blocking-type anti-TSH receptor (TSHR) antibodies using unsolubilized porcine TSHR. Employing this assay, we measured blocking TSH binding inhibitory immunoglobulin (TBII) in the sera from 30 untreated Graves' patients and compared the results with those of a bioassay measuring thyroid stimulation blocking antibodies (TSBAb). Blocking TBII was positive in 9 of 30 sera (30%) and the blocking TBII activity was correlated with the total TBII value, which was measured by the radioreceptor assay using solubilized porcine TSHR. On the other hand, TSBAb determined with the conventional bioassay was positive in only 2 sera (6.7%), and no correlation was observed with the TSAb activity. In some cases in which the TSAb activity was rather high, TSBAb could not be detected by bioassay, whereas blocking TBII was positive. There was no correlation of blocking TBII activity with goiter size, thyroid hormone level, or proptosis. However, there was a tendency for anti-thyroid therapy to require a shorter time for FT4 normalization in blocking TBII subjects, suggesting that blocking-type anti-TSHR antibody plays some role in the pathophysiology of Graves' disease. In conclusion, blocking type anti-TSHR antibodies are often found in the sera of Graves' patients when the blocking-specific radioreceptor assay is applied. PMID- 11873871 TI - Is an elevation in basal follicle-stimulating hormone levels in unexplained infertility predictive of fecundity regardless of age? AB - An elevation in follicle-stimulating hormone (FSH) levels is considered to reflect lowered ovarian function, resulting in poor fecundity in infertile women. However, it remains to be clarified whether or not the significance of FSH levels applies equally to all women irrespective of age. The objective of the present study is to compare basal FSH levels in infertile women who conceived or not after stratification by age. A total of 144 infertile women between ages 25 and 45 who underwent infertility treatment due to unexplained infertility in the University of Tokyo Hospital were included in the retrospective study. Subjects were divided by age into two groups, < 38 (n=98) vs > or = 38 (n=46) years, with ages ranging from 25 to 37, and from 38 to 45, respectively. Blood samples were collected in early follicular phase (day 4-6) for assessment of basal levels of LH, FSH, and PRL. In the older group, pregnant cases had significantly lower FSH levels (6.07 +/- 2.83 mIU/ml) than nonpregnant cases (9.60 +/- 3.67 mIU/ml), whereas no difference in basal FSH levels was observed between pregnant and nonpregnant cases in the younger group. Basal FSH levels of pregnant cases in the older group were significantly lower than those of pregnant cases in the younger group (8.26 +/- 2.95 mIU/ml). Basal LH and PRL levels were not related to fecundity in either group. Thus, an increase in basal FSH levels as a predictor of fecundity should be considered in the context of age. PMID- 11873872 TI - Cross-reactive mechanism for the false elevation of free triiodothyronine in the patients treated with diclofenac. AB - We report three cases of patients exhibiting a false elevation of serum free triiodothyronine (FT3) as a result of a cross-reaction with diclofenac. The first case is a 66-yr-old woman with a long history of rheumatoid arthritis (RA). The patient was receiving diclofenac for the treatment of her RA. The patient was subsequently diagnosed as having thyroid papillary adenocarcinoma and received a subtotal thyroidectomy. After the operation, the patient exhibited postoperative hypothyroidism except for a gradual elevation of FT3. The other two patients also exhibited an elevated serum FT3 level after the administration of diclofenac. Serum FT3 levels in these patients decreased to normal or below normal after diclofenac administration was discontinued. In the first case, the elimination of immunoglobulin from the sera using polyethylene glycol precipitation did not reduce the FT3 level. In our hospital, Vitros ECi (enhanced chemiluminescence enzyme immunoassay) system and Vitros FT3 kit were used for FT3 assay. The patient's FT3 levels were normal or below normal when they were measured using other FT3 kits. FT3 was also detected when diclofenac was dissolved in a phosphate buffered saline. Therefore, we concluded that a cross-reaction between FT3 and diclofenac was the mechanism causing the false elevation of FT3 in these patients. PMID- 11873873 TI - A polymorphism in the promoter region of the glucocorticoid receptor gene is associated with its transcriptional activity. AB - Since glucocorticoid exerts its biological effects by binding to its receptor, the expression efficiency of the glucocorticoid receptor (GR) gene could influence glucocorticoid sensitivity. We found a polymorphism of cytosine/adenine (-22 C/A) in the upstream region of the GR gene. There was no difference in the allelic frequency between normal and type 2 diabetic subjects. The promoter activity determined by luciferase assay was significantly lower in the -22 A allele than in the -22 C allele in both HepG2 (A allele, 4.19 +/- 0.15; C allele, 6.07 +/- 0.27, p < 0.001) and human embryonic kidney 293 cell lines (A allele, 0.93 +/- 0.16; C allele, 1.51 +/- 0.32, p < 0.001). This polymorphism is associated with transcription of the CR gene, which could be related to glucocorticoid sensitivity through an alteration in tissue GR number. PMID- 11873874 TI - Human pharmaceuticals in the aquatic environment a review. AB - There has been increasing concern in recent years about the occurrence, fate and toxicity of pharmaceutical products in the aquatic environment. Many of the more commonly used drug groups (for example antibiotics) are used in quantities similar to those of pesticides and other organic micropollutants, but they are not required to undergo the same level of testing for possible environmental effects. The full extent and consequences of the presence of these compounds in the environment are therefore largely unknown and the issue as a whole is ill defined. Although these compounds have been detected in a wide variety of environmental samples including sewage effluent, surface waters, groundwater and drinking water, their concentrations generally range from the low ppt to ppb levels. It is therefore often thought to be unlikely that pharmaceuticals will have a detrimental effect on the environment. However, the lack of validated analytical methods, limited monitoring data and the lack of information about the fate and toxicity of these compounds and/or their metabolites in the aquatic environment makes accurate risk assessments difficult. PMID- 11873875 TI - Leaching behavior and characteristics of glass components and surrogate nuclides in radioactive vitrified waste forms. AB - Several vitrified waste forms were fabricated and characterized, which contain simulated radioactive waste incineration ash, and a long-term leaching test was conducted by an ISO method for 820 days to assess the chemical durability of vitrified waste forms. Two semi-empirical mechanism models were applied to find out the dominant leaching mechanism of glass elements. For glass elements, dissolution associated with diffusion was the dominant leaching mechanism and leaching characteristics also depend upon solubilities of components. A type of prediction model was applied to observe the long-term leaching behavior of major glass elements and surrogates. Diffusion coefficients and dissolution rate constants, the main parameters in the long-term prediction model, were obtained for glass elements and surrogate nuclides using experimental data for short and long-term periods. The model could be used to predict long-term behavior of such elements to observe and assess the stability of vitrified waste forms. PMID- 11873876 TI - An engineering assessment of the burning of the combustible fraction of construction and demolition wastes in a redundant brick kiln. AB - This paper confirms both technical feasibility and economic potential via the use of redundant brick kilns as an alternative option for disposal of the combustible fractions of construction and demolition wastes by a three-stage analysis. To assess such an idea, one brick kiln was selected for performing an engineering feasibility study. First of all, field sampling and lab-analyses were carried out to gain a deeper understanding of the physical, chemical, and thermodynamic properties of the combustible fractions of construction and demolition wastes. Kinetic parameters for the oxidation of the combustible fractions of construction and demolition wastes were therefore numerically calculated from the weight loss data obtained through a practice of thermogravimetric analyzer (TGA). Secondly, an engineering assessment for retrofitting the redundant brick kiln was performed based on integrating several new and existing unit operations, consisting of waste storage, shredding, feeding, combustion, flue gas cleaning, and ash removal. Such changes were subject to the operational condition in accordance with the estimated mass and energy balances. Finally, addressing the economic value of energy recovery motivated a renewed interest to convert the combustible fractions of construction and demolition wastes into useful hot water for secondary uses. PMID- 11873877 TI - Optimalisation of magnesium ammonium phosphate precipitation and its applicability to the removal of ammonium. AB - Among the physico-chemical abatement technologies, mainly acid scrubbers have been used to control NH3-emission. The disadvantage of this technique is that it yields waste water, highly concentrated in ammonia. In this report, the applicability of the magnesium ammonium phosphate (MAP) process to regenerate the liquid phase, produced by scrubbing NH3-loaded waste gases, was investigated. In the MAP process, ammonium is precipitated as magnesium ammonium phosphate, which can be used as a slow release fertilizer. The influence of a number of parameters, e.g. pH, kinetics, molar ratio NH(+)4/Mg2+/PO(3-)4 on the efficiency of the formation of MAP and on the ammonium removal efficiency was investigated. In this way, optimal conditions were determined for the precipitation reaction. Next to this, interference caused by other precipitation reactions was studied. At aqueous NH(+)4-concentrations of about 600 mg l(-1), ammonium removal efficiencies of 97% could be obtained at a molar ratio NH(+)4/Mg2+/PO(3-)4 of 1/1.5/1.5. To obtain this result, the pH was continuously adjusted to a value of 9 during the reaction. According to this study, it is obvious that the MAP precipitation technology offers opportunities for ammonium removal from scrubbing liquids. The practical applicability of the MAP-process in waste gas treatment systems, however, should be the subject for further investigations. PMID- 11873878 TI - Selection of denitrifying phosphorus accumulating organisms in activated sludge. AB - Although anoxic phosphorus uptake has been demonstrated, the conditions that stimulate or inhibit the anoxic uptake are not fully understood. In this study, the possibility of achieving biological excess phosphorus removal in sequencing batch reactors using nitrate as an electron acceptor without including an aerobic phase has been confirmed. The use of an anaerobic/anoxic/aerobic sequencing batch reactor allows anoxic phosphorus uptake to be achieved, in spite of the low fraction of denitrifying phosphorus accumulating organisms present in the inoculum. Biomass characterisation was carried out to determine the denitrifying fraction of the phosphorus accumulating organisms in sequencing batch reactors operating at different operational conditions. With an anaerobic/anoxic/aerobic cycle, it was demonstrated that the disadvantage of denitrifying phosphorus accumulating organisms due to the lower substrate utilisation efficiency in anoxic conditions relative to aerobic conditions, is not so effective to cause the "strictly aerobic" phosphorus accumulating organisms to dominate in the system. PMID- 11873879 TI - Effect of different operational parameters in the enhanced biological phosphorus removal process. Experimental design and results. AB - The uncontrolled dumping of phosphorus into a water environment creates serious problems of eutrophication, affecting water quality and causing grave problems in the aquatic ecosystem. European legislation demands drastic reduction of phosphorus dissolved in wastewater. Enhanced Biological Phosphorus Removal (EBPR) is the current tendency in wastewater treatment. This biological process depends on a multiplicity of variables, but its three main factors are: influent COD/P ratio, anaerobic fraction and sludge retention time (SRT). The aim of this work is to statistically determine the effect of these three parameters in EBPR through a response surface methodology. The objective function that has been chosen is phosphorus removed per unit biomass formed. This function provides ample information on BPR, since the quantity of phosphorus removed depends on the accumulative capacity of the microorganisms present. Two levels were chosen for the SRT (5 and 10 days), two for the anaerobic fraction (0.1 and 0.2), and six for levels of the influent COD/P ratio (between 16 and 87). The experiments were undertaken at pilot scale (100 litres) with an A2/O configuration, with simultaneous nitrogen and phosphorous removaL The wastewater used is a synthetic mixture of complex sources of carbon and nitrogenwithout volatile fatty acids. The empirical model obtained indicates that the factor most influencing the EBPR process is the influent COD/P ratio, whilst the anaerobic fraction is that which has least influence. Additionally, there is an optimum in the influent COD/P ratio that is to be found between 41 and 48. PMID- 11873880 TI - Removal of arsenite from aqueous solutions by anionic clays. AB - Layered double hydroxides are antitype 2/1 clay minerals that can be synthesized rapidly under laboratory conditions. Due to their high anion exchange capacities, layered double hydroxides have been investigated as potential adsorbents for removal of anionic contaminants from aqueous systems. In this study, uncalcined and calcined layered double hydroxides were prepared and characterized by X-ray diffraction, with the products evaluated for their ability to adsorb As(III) in aqueous solutions. Results indicated that As(III) could be adsorbed on chloride layered double hydroxide and calcined layered double hydroxide, but no adsorption occurred for carbonate layered double hydroxide. The adsorption isotherms of As(III) on chloride layered double hydroxide and calcined layered double hydroxide were typical L and H-type curves, respectively. The adsorption of As(III) on calcined layered double hydroxide was a slow process and reached a quasi-equilibrium after a 20 hr reaction time. The layered double hydroxides had high pH buffering capacities and the As(III) adsorption on calcined layered double hydroxide was a function of pH. Competing anions strongly affected adsorption, with As(III) adsorption increasing in the order: HPO(2-)4 < SO(2-)4 < CO(2-)3 < F- < Cl- < Br- approximately equals I- < NO(-)3. Adsorbed As(III) on calcined layered double hydroxide could be desorbed by different anions, but there was no systematic relationship between As(III) desorption and anion affinities for the calcined layered double hydroxide. Calcination immobilized the As(III) adsorbed on calcined layered double hydroxides. Although layered double hydroxides could be recycled and used as an adsorbent, its adsorption efficiency was reduced with successive treatments. PMID- 11873881 TI - Inhibition on acidogenesis of dairy wastewater by zinc and copper. AB - Acidogenesis of dairy wastewater produced volatile fatty acids, mainly acetate and propionate, plus hydrogen as by-product. Zinc (Zn) inhibited acidogenesis at concentrations over 10 mg l(-1); but at 10 mg l(-1), or less, it enhanced acidogenesis slightly. On the other hand, copper (Cu) inhibited acidogenesis at all tested concentrations ranging 5-400 mg l(-1). Production of acetate was inhibited by both metals at all concentrations; but production of propionate and hydrogen was favored at low concentrations of Zn (up to 80 mg l(-1)) and Cu (up to 40 mg l(-1)). Production of hydrogen corresponded with the degradation of carbohydrate and the production of propionate. Overall, Cu was 1.4-4.3 folds more toxic than Zn, according to the overall production patterns of fatty acids and hydrogen as well as degradation patterns of carbohydrate and protein. PMID- 11873882 TI - Treatment of leachate from a domestic solid waste sanitary landfill by an electrolysis system. AB - This study deals with the characteristisation and the treatability of leachates from the new municipal domestic waste (DW) landfill site in Athens, Greece, using an electrolysis system. This site has been in operation since July 1998. The leachate from this site has mean COD and BOD5 values of 53300 and 30300 mg l(-1), respectively, and constitutes a major environmental problem that must be taken into serious consideration. An electrolysis system, using Ti/Pt electrode in alkaline conditions was used to treat the leachate. The study showed, that the COD was reduced by 84%, and that VSS, N-NH4 and Total Phosphorus were reduced by 100% within one hour of electolysis and at pH 9. The efficiency of electrolysis went up to 32 g COD removed (CODr)/h-A-m2 and the energy consumption was 12.6 kWh kg(-1)CODr. PMID- 11873883 TI - Microbial populations of an upflow anaerobic sludge blanket reactor treating wastewater from a gelatin industry. AB - The microbial populations of an upflow anaerobic sludge blanket reactor, used for treating wastewater from the gelatin industry, were studied by microbiological methods and phase-contrast and electron microscopy. Microscopy examination of the sludge showed a complex mixture of various rod-shaped and coccoid bacterial pluslong filaments and verymobile curved rods. In addition free-living anaerobic ciliates and flagellates were also observed. The trophic group population observed in decreasing order of dominance were hydrolytic and acetogenic at 10(6) and sulfate reducing and methanogenic at 10(5). The rate of methane production in anaerobic granular sludge cultivated in growth medium supplement with formate pressurized with H2:CO2 showed a significant increase in methane yield compared with theseed culture containingthe same substrate and atmosphere of N2:CO2. Similar rates of methane production were observed when the growth medium was supplemented with acetate pressurized either with H2:CO2 or N2:CO2. The number of total anaerobic bacteria at 10(7), fecal coliforms and total coliforms at 10(6), and fecal streptococci at 10(3) is based on colony counts on solid media. The four prevalent species of facultative anaerobic gram-negative bacteria that belong to the family of Enterobacteriaceae were identified as Escherichia coli, Esherichia fergusonii, Klebsiella oxytoca, and Citrobacter freundii. The species Aeromonas hydrophila, Aeromonas veronii, Acinetobacter iwoffi and Stenotrophomonas maltophila were the most frequently isolated glucose fermenting and nonfermenting gram-negative bacilli. PMID- 11873885 TI - Primary care groups as community laboratories. PMID- 11873884 TI - Effect of applying Hong Kong biosolids and lime on nutrient availability and plant growth in an acidic loamy soil. AB - A greenhouse experiment was conducted to evaluate the growth of Brassica chinensis L. in an acidic loamy soil amended with municipal biosolids with or without lime treatment (750 mg kg(-1)). The soil was amended with one of two municipal biosolids from Tai Po (TP) or Yuen Long (YL) wastewater treatment plants at application rates of 0, 5, 10, 25 and 50% (v/v). Initial NH(+)4-N and PO(3-)4-P concentrations in the amended soil increased with an increase in municipal biosolids loading rates and those with biosolids from TP had a greater increase in nutrients than those from YL. However, initial NO(-)3-N contents in both municipal biosolids amended soils decreased with an increase in biosolids loading rates, particularly for the soil amended with YL. Soluble K, Mg, Na and Ca concentrations in the soil increased with application rates. Municipal biosolids amended soil without lime treatment had higher NH(+)4-N, NO(-)3-N and major cation contents than those with lime. Addition of Tai Po municipal biosolids and lime significantly increased the dry weight yields of B. chinensis L., while YL municipal biosolids increased yields only at the 5% rate with lime amendment. The maximum growth was obtained at municipal biosolids application rates of 5 and 10% for TP and 5% for YL for limed soil. Addition of municipal biosolids increased the total N, P, Zn, Cr and major cation concentrations in the shoot tissue, while lime treatment reduced Zn and Cr contents. The high salt and metal concentrations would likely be factors inhibiting plant growth at high application rates, especially for YL biosolids. Further studies to determine the long-term effects of biosolid application on soil and crop quality are warranted. PMID- 11873886 TI - Secular trends in antidepressant prescribing in the UK, 1975-1998. AB - BACKGROUND: We have examined secular trends in age- and sex-specific prescribing of antidepressants to determine whether these mirror changes in other population measures of mental health. METHOD: An analysis was carried out of age- and sex specific rates of antidepressant prescribing by a representative sample or panel of UK general practitioners (GPs) in the period 1975-1998. RESULTS: The number of antidepressant prescriptions issued increased more than twofold in the period 1975-1998 and, in 1998, a total of 23.4 million antidepressant prescriptions were issued by GPs in the United Kingdom. Rates of antidepressant prescribing increased markedly in all age and sex groups with as much as a threefold increase in the older age groups. With the exception of 12-19-year-olds, these increases have been more marked in males, although absolute levels of prescribing are still at least two times higher in females. CONCLUSIONS: Antidepressant prescribing has increased in all age and sex groups. This indicates either that there have been changes in the presentation, recognition and management of depression in general practice or that the prevalence of depression has increased, or a combination of these two phenomena. The higher prescribing rate in females is in keeping with evidence from psychiatric morbidity surveys suggesting that women experience higher levels of psychiatric morbidity than men. Decreases in the ratio of female to male prescribing, however, support other data indicating that, relative to females, the mental health of young males has declined in recent years. Changes in patterns of help-seeking may also contribute to the observed trends. PMID- 11873887 TI - Changes in sports injuries to children between 1983 and 1998: comparison of case series. AB - BACKGROUND: Sports injuries sustained by children are worrying because they prevent and deter participation in physical activity. Before we can address such injuries we need to understand the size of the problem and whether there have been changes in occurrence. A study of sports injuries to children, carried out in a Cardiff Accident and Emergency department in 1983, provided the data against which to compare data gathered in 1998. METHODS: Data on all sports injuries to children aged 16 and under treated between September and December 1998 were compared with those reported for the same hospital, age group, injury and period in 1983. RESULTS: A total of 953 injuries were treated in 1998, representing an increase of 54 per cent [95 per cent confidence interval (CI) 44-64 per cent]. The male:female distribution remained constant and the majority of injuries were due to rugby and soccer. The number of females injured playing rugby and soccer increased and a wider range of sports led to injuries for both males and females. Amongst 10-15-year-olds injury risk increased from 1 in 78 for boys in 1983 to 1 in 22 in 1998 (p < 0.0001). For girls, the increase was from 1 in 117 to 1 in 55 (p < 0.0001). The number of soccer- and rugby-related fractures increased by 52 per cent (95 per cent CI 22-87 per cent). CONCLUSIONS: Sports injury rates have increased considerably over 15 years. With minimal population change, little variation in minor injuries and only small improvements in data capture, the main reason for change appears to be increased participation. PMID- 11873888 TI - Early discharge of low-risk women from cervical screening. AB - BACKGROUND: The Scottish Cervical Screening Programme currently offers three yearly screening to all women between the ages of 20 and 60. However, previous studies have indicated that well-screened women over the age of 50 are likely to be at low risk of cervical neoplasia. This study aimed to explore the implications of discharging these women from screening in a typical area of Scotland. METHODS: A case-control study of the screening histories of women with and without screen-detected cervical neoplasia between ages 50 and 59 in Lanarkshire was carried out, as well as a cross-sectional study of the prevalence of adequate screening histories among women currently aged 50 in Lanarkshire. Routine screening programme statistics were used to estimate the effects of introducing an early discharge policy. RESULTS: Women reaching the age of 50 with two recent, consecutive, negative smears had reduced odds of screen-detected neoplasia in the subsequent decade. The estimated odds ratio for all screen detected neoplasia (CIN 1-3, adenocarcinoma in situ and invasive carcinoma) was 4.4 [95 per cent confidence interval (CI) 1.6-13.2, p = 0.002]. The estimated odds ratio for screen-detected high-grade CIN and invasive squamous carcinoma was 17.0 (95 per cent CI 2.4-243.0, p = 0.0004). A total of 54.0 per cent (95 per cent CI 47.9-59.9 per cent) of screening participants currently aged 50 fulfilled the definition of adequate screening. Discharging these women might be expected to reduce screening workload by approximately 10 per cent, but those discharged would be at increased risk of neoplasia. CONCLUSION: Now that full screening histories are available in all health board areas since 1990, the identification of a low-risk group within the screened population could be the first step towards a screening programme targeted more closely on those with the greatest capacity to benefit. PMID- 11873889 TI - Development and validation of a computerized South Asian Names and Group Recognition Algorithm (SANGRA) for use in British health-related studies. AB - BACKGROUND: Studies on ethnic variations in health have played an important role in aetiological and health services research. Most routine datasets, however, do not include information on ethnicity. South Asians, one of the largest minority ethnic groups in Britain, have distinctive names that also allow differentiation of the main sub-groups with their important differences in health-related exposures and disease risks. METHODS: A computerized name recognition algorithm (SANGRA) was developed incorporating directories of South Asian first names and surnames together with their religious and linguistic origin. SANGRA was validated using health-related data with self-ascribed information on ethnicity. RESULTS: SANGRA was successful in recognizing South Asian origin in reference datasets, with sensitivity of 89-96 per cent, specificity of 94-98 per cent, positive predictive value (PPV) of 80-89 per cent and negative predictive value (NPV) of 98-99 per cent. Religious origin was correctly assigned in the majority of cases: sensitivity, specificity and PPV were 94 per cent, 91 per cent and 90 per cent for Hindus; 90 per cent, 99 per cent and 98 per cent for Muslims; and 76 per cent, 99 per cent and 94 per cent for Sikhs. SANGRA correctly identified 76 per cent Gujerati and 70 per cent Punjabi names, although only 62 per cent of Gujerati names were sufficiently distinct to be allocated to the Gujerati-only category and only 53 per cent Punjabi names were allocated to the Punjabi-only category. However, specificity and PPV were high for both languages (respectively 97 per cent and 93 per cent for Gujerati, and 99 per cent and 97 per cent for Punjabi). CONCLUSIONS: SANGRA provides a practical and valid method of ascertaining South Asian origin by name and, to a lesser degree of accuracy, of differentiating between the main religious and linguistic subgroups living in Britain. This algorithm will be useful in health-related studies where information on self-ascribed ethnicity is not available or is of a limited nature. PMID- 11873890 TI - Hidden need for drug treatment services: measuring levels of problematic drug use in the North West of England. AB - BACKGROUND: In the North West of England, data on drug users are routinely collected from a variety of agencies including specialist treatment centres, police and probation services. However, the covert nature of drug use means that alone, these conventional monitoring systems cannot provide the epidemiology required to target and develop drug treatment and prevention initiatives. METHODS: Utilizing surveillance data and capture-recapture techniques we estimate the rates of problematic drug users by age and sex in five North West health authorities and one local authority. RESULTS: Analyses show concentrations of problematic drug use in large metropolitan areas (Liverpool and Manchester) with levels as high as 34.5 and 36.5 per 1000 population (ages 15-44), respectively, and, for males, levels exceed 50 per 1000 in three authorities. Patterns of prevalence for those aged 25 and over differed from those in the younger age groups, with disproportionate levels of young users outside metropolitan areas. The proportion of young users already in treatment (21.3 per cent) was lower (older users, 35.3 per cent), with overall proportions in treatment varying between health authorities (range 26.2-46.5 per cent). CONCLUSION: With a multi agency approach, established monitoring systems can be used to measure hidden populations of drug users. Estimates of the current populations of such users in the North West of England suggest that planned increases of people in treatment by 100 per cent would fail to accommodate even current level of problematic users. A holistic approach to new initiatives must ensure that the high level of relapse once drug users are discharged are reduced and that the needs of young users are addressed before prolonged treatment is required. PMID- 11873891 TI - Women who are recalled for further investigation for breast screening: psychological consequences 3 years after recall and factors affecting re attendance. AB - BACKGROUND: In 1995-1996 a study was commenced investigating the experience of 'false-positive' women, i.e. who had undergone further investigations following routine breast screening and received a clear final result. These women were found to experience significantly greater adverse psychological consequences at 1 month, 5 months and 11 months after assessment compared with women who received a clear result after the initial basic mammogram. The present study follows up these 'false-positive' women 3 years later (at 35 months) just before being invited for their next routine breast screening. It investigates the effect of the previous experience of breast screening on adverse psychological consequences reported by false-positive women at this time, and explores factors that may be associated with the current adverse psychological consequences. Factors influencing attendance for the forthcoming appointment are reported, and the non attendance rate is monitored. METHODS: Women who had previously completed a questionnaire 1 month, 5 months and 11 months after their last breast screening 3 years ago, were invited to complete a postal questionnaire just before being invited to attend for their next routine mammogram 3 years later. Attendance for this appointment was monitored. A brief questionnaire was sent to non-attenders to ascertain their reasons for not attending. RESULTS: The response rate was 77 per cent (387/505). Women who, at their last routine breast screening, had received a clear result after fine needle aspiration (FNA) at assessment, after a surgical biopsy or after a 6 month early recall appointment, all suffered significantly greater adverse psychological consequences at 1 month before returning for routine breast screening 3 years later than women who had received a clear result after the initial mammogram at their last routine breast screening. They were between 1.7 and 2 times more likely to suffer psychological consequences than women who received a clear result after their last mammogram. Women who had received a clear result at assessment without undergoing FNA reported higher psychological consequences than those who received a clear result after mammography, but the difference was not significant (relative risk 1.28, 95 per cent confidence interval 0.82-2.00). Fifteen per cent of those who had undergone assessment 3 years earlier did not attend their next routine breast screening appointment compared with 8 per cent of those who received a clear result after mammography (p = 0.035). Factors associated with adverse psychological consequences are reported. CONCLUSION: Despite having received a final clear result during their previous routine breast screening 3 years ago, women who had undergone FNA, surgical biopsy or been placed on early recall suffered significantly greater adverse psychological consequences at 1 month before their next routine breast screening appointment than women who had received a clear result after their initial mammogram at their last routine breast screening. Having undergone further investigations did not necessarily motivate women to attend for their next routine appointment, with 15 per cent of these women not returning for routine screening 3 years on. PMID- 11873892 TI - Emergency transfer from independent hospitals to NHS hospitals: risk, reasons and cost. AB - BACKGROUND: In view of public concern about standards of emergency care in independent hospitals and the impact of transferred patients on NHS facilities we aimed to estimate the number and risk of emergency transfers from independent hospitals to NHS hospitals; to describe the circumstances; and estimate costs to the NHS. METHODS: Patients transferred in three months from 137 independent hospitals were identified from central records systems and local hospital enquiries. Circumstances were described by Directors of Nursing in telephone interviews. Numbers were weighted for whole year activity and non-participating hospitals to estimate total transfers in 1999. Medical Directors of NHS Trusts receiving the patients supplied durations of stay in critical care and other facilities. NHS Reference Costs were applied. RESULTS: There were 158 emergency transfers (plus 105 planned transfers, and 18 as a result of funding problems). Proportionately more emergency transfers were from hospitals lacking intensive care facilities. Patients over 65 years old constituted 61 per cent of transfers but only 25 per cent of all cases. Transfer followed major abdominal surgery in 42 (26 per cent) cases and major orthopaedic surgery in 31 (20 per cent), although these treatments constituted only 2 per cent and 3 per cent of the caseloads. There were an estimated 749 emergency transfers in 1999 (95 per cent confidence interval 640-875), a risk of 1 in 956 (all ages) and 1 in 392 (aged over 65); 729 had been funded privately, of whom two-thirds became NHS patients after transfer, costing Pound Sterling 2.61 million. CONCLUSIONS: The scale of emergency transfer (two per day) and resulting cost to the NHS is small. The risk is reducible if patients and interventions are matched to hospitals' critical care capabilities. Common clinical service guidelines should apply to NHS and independent hospitals. PMID- 11873893 TI - A birth cohort analysis of smoking by adults in Great Britain 1974-1998. AB - BACKGROUND: The aim of the study was to determine for Great Britain the percentage of current smokers and ever smokers by age in successive birth cohorts and the percentage of ever smokers who continue, by analysis of data from serial cross-sectional surveys of smoking status (General Household Survey). METHOD: A series of 5 year birth cohorts were followed through data from the 13 national surveys conducted at biennial intervals between 1974 and 1998. RESULTS AND CONCLUSION: In all birth cohorts, male and female, after age 25 years the percentage of current smokers falls with age. In the earliest birth cohort for males (1897-1901) about 85 per cent were ever smokers (i.e. had smoked at some time). After the 1922-1926 cohort this started to fall, to reach the level of about 50 per cent in the 1962-1966 cohort. In females only 25 per cent of the earliest cohort ever smoked but this rose, reaching about 65 per cent in the 1922 1926 cohort before falling back to about 50 per cent in the 1962-1966 cohort. The age at which smokers quit appears to be falling in successive cohorts. Once they have started quitting the rate at which smokers do so is very similar in all cohorts, with about 1 per cent of ever smokers quitting each year. If these trends are continued the UK smoking prevalence targets will not be met. PMID- 11873894 TI - Winter emergency pressures for the NHS: contribution of respiratory disease, experience in North Staffordshire district. AB - The rise in emergency medical admissions in winter in the NHS hospitals in the United Kingdom has been recognized to reflect respiratory and cardiovascular illness. In our study we looked at the contribution of respiratory disease to the winter pressures in our district. Respiratory disease related emergency admissions increased twofold in the winter months, with obvious implications for workload. An evidence-based National Service Framework for respiratory disease would be useful. PMID- 11873895 TI - The onset of the excess of childhood cancer in Seascale, Cumbria. AB - BACKGROUND: The objective of this study was to formally investigate the onset of the Seascale cluster of childhood and young person's cancer. This has not previously been attempted. METHODS: A mortality study within the Whitehaven registration district was set up and death records were abstracted for 1906-1970. They were categorized as death from leukaemias, lymphomas, other cancers and all other causes in persons aged 0-14, 0-24 and 25-84. The number of deaths, death rates and standardized mortality ratios were calculated. RESULTS: The mortality of persons aged 25-84 in Seascale civil parish, Gosforth civil parish and the rest of the Whitehaven district was unremarkable compared with national data 1906 1970. There were no cancer deaths aged 0-24 in Gosforth civil parish during 1906 1970. In Seascale civil parish a hitherto unrecorded childhood cancer case was revealed, dying in 1954. No cancer deaths aged 0-24 were found before that date. In the period 1946-1955 three cancer deaths gave a statistically significant excess owing to non-leukaemia cases, whereas in the period 1956-1965 a statistical excess of all types of leukaemia occurred as a result of two deaths. There was no case excess (based on one leukaemia death) in the period 1966-1970. CONCLUSION: We found no clear temporal associations of the case excesses either with the periods of significant nuclear activity on the Sellafield site or with the main periods of population growth in the area. PMID- 11873896 TI - Retirement intentions of doctors who qualified in the United Kingdom in 1974: postal questionnaire survey. AB - BACKGROUND: Medical workforce planning needs to be informed by knowledge about doctors' retirement intentions. Systematic information about retirement intentions, and factors that influence them, is sparse. METHODS: Postal questionnaires were sent to members of a cohort of medical qualifiers surveyed regularly since they qualified in 1974, with quantitative analysis of intentions about early retirement and qualitative analysis of reasons for wanting early retirement. RESULTS: A total of 1717 replies were received from 2217 traceable doctors (77.4 per cent). Of these, 1427 doctors worked in the NHS and answered the question about retirement: 14.8 per cent (211) said that they would definitely continue to normal retirement age and 20.1 per cent (287) probably would. Of those not definitely continuing to normal retirement age, 45.1 per cent had made financial provision to support early retirement, Seventy per cent cited reasons for considering early retirement: the main reasons were to reduce work related pressure, increase leisure time, job dissatisfaction, disillusionment with the NHS, and wanting a healthy retirement. Doctors might be encouraged to stay by more flexible working patterns, a reduction in workload with increasing age, improved staffing levels, preservation of pension rights for part-time working, fewer NHS administrative changes, and greater professional freedom. CONCLUSION: The impact of early retirement on medical workforce supply may be considerable. Approaches to retirement policy need to shift away from the extremes of either full-time employment or total retirement. PMID- 11873897 TI - Obstacles to influenza immunization in primary care. AB - BACKGROUND: General practices undertake annual immunization campaigns to protect susceptible patients against influenza. Many practices, however, do not adopt effective approaches and there is great variation in the immunization rates achieved. This study aimed to assess the attitudes of primary care staff to the annual immunization programme, the obstacles they face, and possible reasons for the wide variation in immunization rates. METHOD: A semi-structured questionnaire survey of general practice groups in Salford &Trafford during winter 1997-1998 was carried out, a total of 104 practices. RESULTS: Respondents perceived influenza vaccine to be effective (93.2 per cent), well received by patients (91.7 per cent) and without significant side-effects (83.6 per cent). The annual immunization programme was seen as being necessary (91.8 per cent), cost effective (76.7 per cent), reducing hospital admissions (82.2 per cent), but very time consuming (64.4 per cent). Practices were more likely to target patients specified in the Chief Medical Officer's guidelines; however, most (98.6 per cent) targeted the over-75s before their inclusion in the guidelines, and many (61.6 per cent) targeted the over-65s. Practices did not always use the most effective methods of contacting patients, primarily relying on posters (97.3 per cent), opportunistic contacts (95.9 per cent) and reminders on prescriptions (83.6 per cent), rather than letters (39.7 per cent) and telephone calls (11.0 per cent). Practices identified several common obstacles to immunization, relating to the cost and administrative burden of the annual immunization programme, difficulty identifying high-risk patients, and public beliefs about influenza and influenza vaccine. CONCLUSIONS: Although generally positive towards influenza immunization, practice staff differ in their ability to cope with, and the appropriateness of their response to, the pressures of the annual immunization programme. Additional support and co-operation from the Department of Health, Health Authorities and pharmaceutical companies could remove some of the obstacles to immunization of high-risk patients. PMID- 11873898 TI - A study of neonatal BCG immunization within an acute hospital trust. AB - BACKGROUND: Tuberculosis is a re-emerging problem in the United Kingdom. BCG immunization administered in the neonatal period is protective. National guidelines and locally published standards identify infants for whom BCG immunization is recommended. The study aimed to calculate the rate of identification of infants 'at risk' by parental ethnic group and/or family history of tuberculosis, to determine subsequent immunization uptake, and to describe characteristics associated with missed BCG immunization. METHODS: A retrospective audit was conducted. Demographic data were collected from a computer database of antenatal booking data, for 2043 pregnancies delivering between 1 October 1998 and 30 April 1999. A cohort of infants 'at risk' was defined, and infants referred for BCG immunization were identified. A manual search of immunization records determined immunization uptake. RESULTS: A cohort of 247 (12 per cent pregnancies) was 'at risk'. Fifty-five per cent of the cohort 'at risk' was correctly identified and 42 per cent correctly identified and immunized. The largest subgroup of the cohort, 48 per cent, was Caucasian and at risk because of a positive family history of tuberculosis. Family history of tuberculosis was the most important risk factor, and was missed in 86 per cent of cases. CONCLUSIONS: Despite the local publication of established guidelines, 58 per cent of infants 'at risk' failed to be immunized. Family history of tuberculosis was more important than parental ethnic group in predicting risk for the cohort, and was missed in the majority of cases. Appropriate guidelines alone do not guarantee good practice. Guidelines should be introduced in conjunction with regular audit to ensure effective implementation. PMID- 11873899 TI - Examining the direct costs and effectiveness of syphilis detection by selective screening and partner notification. AB - Selective screening and partner notification are two principal means of preventing and controlling syphilis in the United States, yet few studies have been undertaken to compare and evaluate the cost or effectiveness of detecting syphilis using either strategy. The objective of this paper is to assess from the perspective of a health department the cost-effectiveness of selective screening compared with the strategy of partner notification in the detection of early syphilis in Houston, Texas, in 1994 and 1995. The cost-effectiveness analysis was performed using the recurring direct costs associated with detecting syphilis by both strategies. The middle estimates for the total direct costs associated with selective screening and partner notification were $579,101 and $229,529, respectively, for the 1466 and the 567 cases of early syphilis detected. On a cost per case basis, selective screening was more cost-effective than partner notification in the detection of primary, secondary and maternal syphilis cases. However, when consideration was given to prophylactic treatment, partner notification was more cost-effective in the detection of all early stage disease. Our findings suggest that the relative benefit of partner notification over selective screening depends on prophylactic treatment and an increase in worker productivity. PMID- 11873900 TI - Promoting uptake of influenza vaccination among health care workers: a randomized controlled trial. AB - In a randomized controlled trial, an intensive promotional campaign failed to increase the uptake of vaccination against influenza among health care workers. The uptake of vaccination was low. PMID- 11873901 TI - Experience of 'screening' for domestic violence in women's services. PMID- 11873902 TI - Quarterly communicable disease review. April to June 2001. PMID- 11873904 TI - Audit of clinical nurse practitioner led thrombolysis. PMID- 11873903 TI - Trends in antibiotic prescribing. PMID- 11873905 TI - My favourite software. PMID- 11873906 TI - Specific and global regulation of genes associated with the degradation of aromatic compounds in bacteria. AB - A large number of bacteria are able to degrade aromatic carbon sources employing different strategies. All these pathways are objects of regulatory control at the level of gene expression. This includes specific control in response to the availability of the respective substrate and in many cases global control responding to other available carbon sources or to the metabolic status of the cell. Here, the regulatory proteins responsible for gene regulation are reviewed in particular in correlation to other proteins with a similar primary structure. Most common is the appearance of regulators of the LysR family; other abundant regulator types are NtrC/XyIR-type proteins, AraC/XyIS-type proteins and the IcIR type proteins. Almost all of the regulators exert their effects as activators of gene expression with the exception of the GntR-type proteins, which are exclusively described as repressors. Factors involved in individual cases of global regulatory mechanisms are enterobacterial CAP, (p)ppGpp, Crc protein, and direct modification of a specific regulator. However, for most pathways of aromatic compound degradation, the molecular mechanisms causing global regulation are not understood. PMID- 11873907 TI - Improved quantitation and reproducibility in Mycobacterium tuberculosis DNA microarrays. AB - We show here that the amount of labeled cDNA and its specific activity can play a significant role in the quantitation of microarray experiments. Standard reverse transcription of 2 microg total bacterial RNA with concomitant incorporation of cyanine dye-conjugated nucleotides did not produce enough label for optimal hybridization results in our Mycobacterium tuberculosis DNA microarray. Therefore we turned to an alternative labeling method using the incorporation of aminoallyl nucleotides followed by conjugation to Cy-dye. The method allows up to 10 fold more label to be produced, and at higher specific activity. In particular, more transcripts can be detected and variability between replicate features can be reduced by using more labeled cDNA. We show that optimizing the labeling protocol is a critical element in conducting microarray experiments and obtaining reproducible and interpretable data. PMID- 11873908 TI - A vector with transcriptional terminators increases efficiency of cloning of an RNA virus by reverse transcription long polymerase chain reaction. AB - Full-length cDNA clones of RNA viruses are advantageous for maintaining the genomic sequence without the generation of diversity by accumulation of sequence mutations during productive virus replication. They permit in vitro manipulation of the genomic clone to test the effect of sequence changes on the phenotype of reactivated virus. Infectious cDNA clones have been produced by ligation of subgenomic clones but are sometimes difficult to generate in a single cloning operation. We used reverse-transcription to synthesize full-length cDNA from genomic RNA of Coxsackievirus B3 of the Picornavirus family and enzymatically amplified this by long PCR. Five different cloning vectors were used to clone the long PCR product, including the vector Lorist6 which contains transcriptional terminators on either side of the cloning site to prevent transcription of inserts in E. coli. No recombinant colonies were obtained from any of the vectors lacking transcriptional terminators but three full-length clones were obtained using Lorist6. The results suggest that transcriptional terminators increase the recovery of cDNA clones of the 7.4 kb Coxsackie virus genome in this cosmid vector, without resort to phage packaging, representing an advance over previous methods and advantages in the molecular manipulation of these viruses. PMID- 11873909 TI - Sequence similarity as a predictor of the transmembrane topology of membrane intrinsic subunits of bacterial respiratory chain enzymes. AB - Integral membrane proteins usually have a predominantly alpha-helical secondary structure in which transmembrane segments are connected by membrane-extrinsic loops. Although a number of membrane protein structures have been reported in recent years, in most cases transmembrane topologies are initially predicted using a variety of theoretical techniques, including hydropathy analyses and the "positive inside" rule. We have explored the use of plots of the distribution of sequence similarity within families of membrane proteins comprising homeomorphic domains as a new method for the prediction/verification of the orientation of transmembrane topology models within certain families of multimeric respiratory chain enzymes. Within such proteins, analyses of sequence similarity can: i) identify heme and/or quinol binding sites; ii) identify potential electron transfer conduits to/from prosthetic groups; and iii) locate regions defining potential subunit-subunit interactions. We mined emerging bioinformatic data for sequences of 11 families of membrane-intrinsic proteins that are part of multimeric respiratory chain complexes that also have membrane-extrinsic subunits. The sequences of each family were then aligned and the resultant alignments converted into a graphical format recording an empirical measure of the sequence similarity plotted versus residue position. In each case, this plot was compared to the predicted transmembrane topology. With one exception, there is a strong correlation between the existence PMID- 11873910 TI - Analysis of regulatory elements and genes required for carbon tetrachloride degradation in Pseudomonas stutzeri strain KC. AB - Previously, we described the generation and initial characterization of four Tn5 mutants of Pseudomonas stutzeri strain KC with impaired ability to degrade carbon tetrachloride (Sepulveda-Torres et al., 1999). In this study, we show cloning and sequencing of an 8.3 kbp region in which all four transposons were located. This fragment encodes eight potential genes and is located in the central part of the 25 kbp fragment recently identified by Lewis et al. (2000) and shown by them to be sufficient to confer carbon tetrachloride transformation capability upon other pseudomonads. The four transposon insertion mutants mapped in ORF's F and I designated by Lewis et al. (2000). This is consistent with the results by Lewis et al. (2000) that orfFis required for carbon tetrachloride degradation. We further established that orfl is required for CCl4 degradation since the three mutants in this ORF were unable to degrade carbon tetrachloride. We present our analysis of the gene and protein sequences from the 8.3 kbp region and propose a tentative model for the role of different genes in the synthesis and activity of pyridine-2,6-bis(thiocarboxylate) (PDTC), the secreted factor responsible for carbon tetrachloride dechlorination. We also found a putative promoter that overlaps with a Fur-box-like sequence in the region upstream of mutated genes. To test this putative promoter region and Fur-box, we generated and ligated DNA fragments containing wild-type and mutant Fur-boxes to a lacZ reporter. The wild type fragment showed promoter activity that is regulated by the concentration of iron in the medium. Finally, we screened a selection of Pseudomonas strains, including P. putida DSMZ 3601--a strain known to produce PDTC--for the presence of the genes characterized in this study. None of the strains tested positive, suggesting that Pseudomonas stutzeri strain KC may possess a distinct biosynthetic pathway for PDTC production. PMID- 11873911 TI - Characterization of the ves gene, which is expressed at a low temperature in Escherichia coli. AB - A gene, designated ves, that is expressionally responsive to temperature was found in Escherichia coli. Experiments with a single-copy lacZ operon fusion and primer extension analysis revealed that ves was expressed at a low temperature with a peak around 25 degrees C but was hardly expressed at 42 degrees C. After a temperature downshift, the mRNA level increased until 6 to 12 h and then decreased. Consistently, an A + T-rich sequence similar to UP elements seen in cold-shock inducible cold-shock protein (Csp) genes was found up-stream of the ves promoter, and its 5'-untranslated region was found to share similarity with those of the cold-shock inducible and cold-adaptive cspA and cspB genes. Additionally, a putative down-stream box, which also exists in cold-inducible proteins, was found. The ves product was identified by overproduction and determination of its N-terminal sequence. Similarity of the C-terminal portion of Ves to the CspA family suggests that Ves belongs to this family. The results of gene-disruption experiments suggest that ves is not essential for E. coli. PMID- 11873912 TI - Questions of interest. PMID- 11873913 TI - Baseline staging tests for breast cancer. PMID- 11873914 TI - Safer injection facilities for injection drug users: the debate continues. PMID- 11873915 TI - Safer injection facilities for injection drug users: the debate continues. PMID- 11873916 TI - Safer injection facilities for injection drug users: the debate continues. PMID- 11873917 TI - Safer injection facilities for injection drug users: the debate continues. PMID- 11873918 TI - Interplanetary health care report cards. PMID- 11873919 TI - The staff and the "fiery serpent". PMID- 11873921 TI - Emergency department overcrowding: ambulance diversion and the legal duty to care. PMID- 11873920 TI - The effect of low-intensity pulsed ultrasound therapy on time to fracture healing: a meta-analysis. AB - BACKGROUND: The effect of low-intensity ultrasonography on fracture healing is controversial, and current management of fractures does not generally involve the use of ultrasound therapy. We describe a systematic review and meta-analysis of randomized controlled trials of low-intensity pulsed ultrasound therapy for healing of fractures. METHODS: We searched 5 electronic databases (MEDLINE, EMBASE, Cochrane Database of Randomised Clinical Trials, HealthSTAR and CINAHL) for trials of ultrasonography and fracture healing, in any language, published from 1966 to December 2000. In addition, selected journals published from 1996 to December 2000 were searched by hand for relevant articles, and attempts were made to contact content experts in the area of ultrasound therapy and fracture healing as well as primary authors of reviewed trials. Trials selected for review met the following criteria: random allocation of treatments; inclusion of skeletally mature patients of either sex with 1 or more fractures; blinding of both the patient and the assessor(s) as to fracture healing; administration of low intensity pulsed ultrasound treatments to at least 1 of the treatment groups; and assessment of time to fracture healing, as determined radiographically by bridging of 3 or 4 cortices. Two reviewers independently applied selection criteria to blinded articles, and selected articles were scored for methodologic quality. The internal validity of each trial was assessed with the use of a 5 point scale that evaluates the quality of trial method on the basis of description and appropriateness of randomization and double-blinding, and assessment of study withdrawals and likelihood of bias. RESULTS: We identified 138 potentially eligible studies, of which 6 met our inclusion criteria. Agreement beyond chance of quality assessments of the 6 trials was good (intraclass correlation coefficient 0.77, p = 0.03). One trial was a repeat analysis of previously reported data, and 2 trials appeared to report on a shared group of subjects. Three trials, representing 158 fractures, were of sufficient homogeneity for pooling. The pooled results showed that time to fracture healing was significantly shorter in the groups receiving low-intensity ultrasound therapy than in the control groups. The weighted average effect size was 6.41 (95% confidence interval 1.01-11.81), which converts to a mean difference in healing time of 64 days between the treatment and control groups. INTERPRETATION: There is evidence from randomized trials that low-intensity pulsed ultrasound treatment may significantly reduce the time to fracture healing for fractures treated nonoperatively. There does not appear to be any additional benefit to ultrasound treatment following intramedullary nailing with prior reaming. Larger trials are needed to resolve this issue. PMID- 11873922 TI - Dances with the pharmaceutical industry. PMID- 11873923 TI - The deferiprone controversy: time to move on. PMID- 11873924 TI - Early Toronto experience with new standards for industry-sponsored clinical research: a progress report. PMID- 11873925 TI - Physicians' liability and drug formulary restrictions. PMID- 11873926 TI - The legal duty of physicians and hospitals to provide emergency care. AB - Accessibility of hospital emergency services has been an issue of increasing concern and was recently brought into public focus in Ontario by the tragic death of Joshua Fleuelling, whose ambulance was redirected from the nearest hospital. As will be reviewed, the limited case law has identified a legal duty for physicians and hospitals to provide treatment to people in need of emergency care, a duty that should be considered when formulating hospital policies. The impact of this duty of care on the existing standard of medical practice will be considered. PMID- 11873927 TI - Nephrology: 3. Safe drug prescribing for patients with renal insufficiency. PMID- 11873928 TI - Can you get tuberculosis twice? PMID- 11873929 TI - Irritable bowel syndrome: could it be celiac disease? PMID- 11873930 TI - Epoetin alfa (Eprex): reports of pure red blood cell aplasia. PMID- 11873931 TI - The Olivieri dispute: no end in sight? PMID- 11873932 TI - Canada's first rural medical school: is it needed? Will it open? PMID- 11873933 TI - Australia's first rural medical school prepares to graduate first MDs. PMID- 11873934 TI - BC takes ax to budget of a health system "in danger". PMID- 11873935 TI - Ontario defies US firm's genetic patent, continues cancer screening. PMID- 11873936 TI - Talking about errors instead of hiding them goal of Vancouver hospital. PMID- 11873937 TI - Proportion of private sector spending declining. PMID- 11873938 TI - Catechol-O-methyltransferase and Parkinson's disease. AB - Parkinson's disease (PD) is one of the main causes of neurological disability in the elderly. Levodopa is the gold standard for treating this disease, but chronic levodopa therapy is complicated by motor fluctuation and dyskinesia. The catechol O-methyltransferase (COMT) inhibitors represent a new class of antiparkinsonian drugs. When coadministered with levodopa/decarboxylase inhibitor, 2 COMT inhibitors, tolcapone and entacapone have been shown to improve the clinical benefit of levodopa. COMT activity is genetically polymorphic, and individuals with the low activity (COMT(L/L)) genotype have a thermolabile COMT protein; studies suggest that this genotype is less common in Asians than in Caucasians. Differences in COMT activity may determine the individual response to levodopa and result in ethnic differences in PD susceptibility. Our recent study suggests that the COMTL allele can interact with the MAOB gene to increase the occurrence of PD in Taiwanese. In order to understand this new class of antiparkinsonian drugs, we review their basic properties, pharmacology, and clinical efficacy. The frequency distribution of COMT genetic polymorphisms among different populations and its implications in the etiology and drug response is also discussed. PMID- 11873939 TI - Neoadjuvant treatment with docetaxel and the effects of irradiation for human ovarian adenocarcinoma and cervical squamous cell carcinoma in vitro. AB - The in vitro radiosensitizing effects of docetaxel have been reported, but the DNA damage caused by the irradiation after docetaxel exposure has not been investigated. In this study, the authors attempted to evaluate the radiosensitizing effects in terms of cell survival and DNA single-strand breaks in a human ovarian adenocarcinoma cell line (known as line BG-1) and a human cervical squamous cell carcinoma cell line (known as line SiHa). The cell lines were exposed to various concentrations of docetaxel (from 2.27 x 10(-3) to 2.27 microg/ml) to investigate the cytocidal effects by colony-formation assay. DNA single-strand breaks after exposure to 2.27 microg/ml of docetaxel for 30 min or 100 min were measured by the alkaline-elution assay. The remarkable cytotoxicity of docetaxel followed by irradiation was observed when concentrations were greater than 2.27 x 10(-2) microg/ml in both cell lines. The combination of docetaxel and irradiation appears to be supraadditive. The DNA single-strand breaks induced by the irradiation were enhanced in both cell lines (BG-1; P < 0.01, SiHa; P < 0.05). The synergistic cytocidal effect cannot be explained quantitatively only by the single-strand breaks. PMID- 11873940 TI - The effect of immobilization stress on the pharmacokinetics of omeprazole in rats. AB - The effects of immobilization stress on the pharmacokinetics of omeprazole were studied in rats. The immobilization stress for 30 or 60 min immediately after oral administration of the drug caused an increase in the time to reach the maximum concentration. However, such stress did not alter the area under the plasma concentration-time curve (AUC). When administered intravenously, the half life during the elimination phase was significantly prolonged by 30 min of immobilization stress, but the AUC value remained unchanged. The intestinal propulsive activity was significantly decreased by immobilization stress. These findings suggest that immobilization stress reduces gastrointestinal motility. A resulting delay during the absorption phase of omeprazole occurs, although the degree of influence on overall pharmacokinetics is relatively insignificant. PMID- 11873941 TI - Human BRAL1 and BCAN genes that belong to the link-module superfamily are tandemly arranged on chromosome 1q21-23. AB - We herein determined by fluorescence in situ hybridization the chromosomal localization of 2 human genes, BRAL1 and BCAN, both of which belong to the link module superfamily, i.e. to the same band of chromosome 1q21-23. Further analysis of the genomic organization of BRAL1 and BCAN revealed that the BRAL1 gene was located 20-kb upstream of the BCAN start site. We isolated a polymorphic dinucleotide (CA) repeat sequence from a genomic clone containing the BCAN gene. High heterozygosity (0.79) makes this polymorphism a useful marker in the study of genetic disorders. Knowledge of the structure of the genes and the marker provides essential information for further analysis of the gene locus at chromosome 1q21-23. PMID- 11873942 TI - Localization of S100C immunoreactivity in various human tissues. AB - Using 2-dimensional gel electrophoresis, we previously demonstrated that the S100C protein remarkably decreased after immortalization of normal human fibroblasts, and that this protein caused growth inhibition of human tumor cells when forcibly expressed in these cells, suggesting that S100C plays a significant role in tumor suppression. The present study was carried out to determine what type of human tissues express S100C protein, and, subsequently, whether the S100C content in these tissues changes after normal cells have been transformed into cancer cells. We found that ductal cells in various tissues were positively stained with the S100C protein. In comparison, epithelial cells in digestive organs such as the stomach, small intestine, and colon were not stained as strongly. When 14 pairs of human normal and cancerous tissues were stained with the antibody, decreases in the staining levels of S100C were observed in 6 kinds of cancerous tissues--from the bronchus, mammary duct, renal tubule, prostate, uterus, and testis--in comparison with staining in their normal counterparts. These results suggest that S100C is a new tumor marker protein, the expression of which significantly decreases after malignant transformation of human tissues. PMID- 11873943 TI - Enhancement of gene transduction efficiency in cancer cells using cationic liposome with hyperthermia. AB - We evaluated the effects of hyperthermia on the efficiency of gene transduction by using a cationic liposome to develop an efficient method for lipofection. We used Lewis lung carcinoma (LLC), NIH3T3, and A549 cell lines, with Lipofectamine reagent as the cationic liposome and the LacZ gene as the reporter gene. In LLC, co-incubation of the cationic liposome and plasmid DNA complex (lipoplex) with the cells for 2 h at 41 degrees C enhanced the efficiency of gene transduction approximately 1.4-fold compared to incubation for 2 h at 37 degrees C, as measured by X-gal staining and beta-galactosidase activity. In cell lines NIH3T3 and A549, the efficiency of gene transduction showed a tendency toward enhancement after 2 h co-incubation with lipoplex at 41 degrees C compared to that at 37 degrees C, as measured by X-gal staining. This is the first study to demonstrate the enhancement of gene transduction efficiency achieved by using a cationic liposome under conditions of hyperthermia. This method should prove useful for lipofection in other cancer cells. PMID- 11873944 TI - Cytoskeletal inhibitors, anti-adhesion molecule antibodies, and lectins inhibit hepatocyte spheroid formation. AB - We investigated the role of cytoskeletons, adhesion molecules, membrane glycosylations, and proteoglycans in forming the shape of adult rat hepatocyte spheroids. Isolated hepatocytes were cultured on dishes coated with chondroitin sulfate phosphatidyl ethanolamine (CS-PE). Spheroid-forming ability was observed after adding cytoskeletal inhibitors (cytochalasin D, colchicine, okadaic acid, mycalolide B), anti-adhesion molecule antibodies (anti-E-cadherin, anti-connexin 32, anti-zo-1), a glycosphingolipid synthetic inhibitor (N butyldeoxynojirimycin), a proteoglycan synthetic inhibitor (p-nitrophenyl-beta-D xylopyranoside), and several lectins. Localization of actin was studied using confocal microscopy after rhodamine-phalloidin staining. Adding cytoskeletal inhibitors on the initial day resulted in weakly clustered cell aggregates rather than smoothly formed spheroids. These effects disappeared at lower reagent concentrations. When reagents were added on day 3, after the formation of spheroids, only mycalolide B was associated with an irregular spheroid surface; the others had no effect. Adding the anti-E-cadherin, anti-connexin 32 on the initial day showed inhibition of spheroid formation, but anti-zo-1 and proteoglycan synthetic inhibitor had no effects. Among the several lectins, only Wheat Germ Agglutinin (WGA), Ricinus communis Agglutinin I (RCA-I), and Concanavalin A (ConA) showed inhibition. These results suggest that cytoskeletal conformation and some adhesion molecules are necessary to form spheroids. Based on the interactions between lectins and hepatocytes in the present study, hepatocytes appear to contain an N-linked complex or N-linked hybrid glycosylated chains. PMID- 11873945 TI - Laparoscopic radical prostatectomy: initial cases at Okayama University Hospital. AB - We performed laparoscopic prostatectomy in seven cases with organ-confined prostate cancer. In 6 cases, the surgery was completed successfully and the mean operative time was 424 min. Volume of blood loss was 200 to 3,200 ml and catheterization lasted 6 to 37 days. No major complications were observed in 6 of the cases. In one case, open surgical conversion was necessary mainly due to a bladder injury. Although these were the first cases of laparoscopic prostatectomy in our institution, the technical difficulty and complexity of the surgery were moderate. We believe that laparoscopic radical prostatectomy will become a standard option for the treatment of organ-confined prostate cancer. PMID- 11873946 TI - A case of suffocation by an advertising balloon filled with pure helium gas. AB - We encountered a rare case of suffocation by an advertising balloon filled with pure helium gas. Suffocation caused by inhalation of atmosphere lacking in oxygen is not exceptional, but reports of death by suffocation due to a pure inert gas such as helium are very rare. In this case, the balloon mooring on the ground was enclosed, warning signs were displayed, and it was clear that entering the balloon filled with an atmosphere lacking in oxygen was extremely dangerous and should not be done; the accident did, however, occur. Accidents of this kind may occur in the future unless appropriate education and countermeasures are taken. PMID- 11873947 TI - The clinical value of urinary N-acetyl-beta-D-glucosaminidase levels in childhood age group. AB - N-acetyl-beta-D-glucosaminidase is a high molecular-weight lysosomal enzyme found in many tissues of the body. It cannot pass into glomerular ultrafiltrate due to its high molecular weight. However, this enzyme shows high activity in renal proximal tubular cells, and leaks into the tubular fluid as the ultrafiltrate passes through proximal tubules. When proximal tubular cells are injured due to to any disease process including glomerular proteinuria, nephrolithiasis, hyperglycemia, interstitial nephritis, transplant rejection or nephrotoxic agents such as antibiotics, antiepileptics, or radiocontrast agents, its urine level increases and thus is used as a reflection of proximal tubular cell necrosis. However, the clinical use of urinary N-acetyl-beta-D-glucosaminidase determination is limited in childhood because of certain technical problems. In addition, the urinary level of this enzyme changes with the maturational level of proximal tubular cells. Thus, difficulties are involved in assessing normal urine levels of this enzyme for age. On the other hand, successive measurements of urinary N-acetyl-beta-D-glucosaminidase during the longitudinal follow-up of the patients may enhance its clinical use as an indicator of ongoing tubular injury. PMID- 11873948 TI - Liver research group considers vascular hepatic diseases. PMID- 11873949 TI - What is your diagnosis? Hypoadrenocorticism (Addison's disease). PMID- 11873950 TI - Feline hyperthyroidism: advances towards novel molecular therapeutics. AB - Feline hyperthyroidism is the most common endocrine disorder of the elderly cat. Traditionally, the disease is treated by surgical thyroidectomy, medical management with antithyroid drugs or radiation therapy using iodine-131. However, none of these treatments is ideal and molecular therapeutics may offer novel methods of treating the disease. This article reviews the background of, and preliminary investigations into, the development of a transcriptionally targeted somatic gene therapy strategy for the treatment of this feline condition. PMID- 11873951 TI - Influence of a high fibre diet on glycaemic control and quality of life in dogs with diabetes mellitus. AB - A study was undertaken to evaluate a high fibre diet used in the management of 10 dogs with naturally occurring insulin-dependent diabetes mellitus. Following baseline measurements of health and glycaemic control, the dogs were fed a canned diet containing a blend of insoluble and soluble dietary fibres and were monitored during the ensuing four months. Switching to the high fibre diet was associated with significantly lower mean 24-hour and postprandial plasma glucose concentrations, which were maintained over the study period. The high fibre diet was also associated with significant reductions in plasma concentrations of fructosamine, glycated haemoglobin, free glycerol and cholesterol, and there were significant improvements in dog activity and demeanour. Bodyweight declined during the fourth month of feeding the diet, which is likely to have resulted from underfeeding relative to increased activity. The results indicate that a high fibre diet can significantly improve glycaemic control and quality of life in dogs with diabetes mellitus. PMID- 11873952 TI - Canine leucocyte adhesion deficiency in Irish red and white setters. AB - Seventy-six Irish red and white setter samples were tested for the recently documented CD18 point mutation which manifests as canine leucocyte adhesion deficiency (CLAD) in Irish setters. Six carrier dogs were identified, all originating from a lineage within which sporadic deaths from symptoms consistent with CLAD had been observed. This is the first demonstration that the CLAD mutation exists outside the Irish setter population, confirming that the mutation is present in a significant minority of Irish red and white setters. Routine testing by breeders to identify carrier animals will enable the eradication of CLAD from the Irish red and white setter population. PMID- 11873953 TI - Neosporosis with cerebellar involvement in an adult dog. AB - A case of an adult dog with multifocal, progressive neurological signs caused by Neospora caninum is reported. Pathological studies showed cerebellar lesions due to the parasite, which was also present in other parts of the nervous system and muscle. Cerebellar atrophy related to Neospora infection has been rarely reported in veterinary medicine, and has been shown to affect ruminants and dogs. The cerebellar involvement and the age of the present dog make this case uncommon. PMID- 11873955 TI - Incomplete humeral condylar fractures in the dog. PMID- 11873954 TI - Balloon dilation for the treatment of chronic recurrent nasopharyngeal stenosis in a cat. AB - Balloon dilation was used for the treatment of a recurrent nasopharyngeal stenosis in a cat with chronic stertorous breathing. The procedure was performed on three occasions at intervals of three and four months using a 15 mm valvuloplasty balloon dilation catheter. After the first dilation, clinical signs reappeared within two weeks due to re-stenosis. After the second dilation, all signs of nasopharyngeal disease disappeared, and did not recur. Endoscopic re evaluation after four months revealed a markedly enlarged nasopharyngeal opening with some degree of stenosis, and a third balloon dilation was performed. Five months later, the cat remained without clinical signs, although some recurrence of the stenosis was visible endoscopically. Complications occurred after the first dilation, and included mild conchal necrosis and transient mucopurulent discharge. Balloon dilation is a minimally invasive technique and can be a successful procedure for treating nasopharyngeal stenosis in cats. PMID- 11873956 TI - Veterinary education in 2010 and beyond. PMID- 11873957 TI - From broken bones to aches and limps at Congress 2002. PMID- 11873958 TI - Towards a DNA diagnosis for endemic arthropod-borne infections. PMID- 11873959 TI - Determination of triorganotin species in water samples by liquid chromatography electrospray-mass spectrometry. AB - Liquid chromatography coupled to electrospray-mass spectrometry (LC-ES-MS) with positive ion detection was evaluated for the determination of tributyltin and triphenyltin in water samples using tripropyltin as internal standard. The separation was performed in the isocratic mode on a silica-based C18 column with a mobile phase containing 0.02% trifluoroacetic acid in acetonitrile-water (50:50, v/v). The optimum LC-ES-MS conditions were established and quantification was performed on the basis of the [M]+ ions. Limits of detection for standard solutions were 100 and 200 pg Sn injected for triphenyltin and tributyltin, respectively, and good reproducibility was observed. Solid-phase extraction was carried out on C18 cartridges to preconcentrate the analytes from natural water samples, with recoveries ranging from 80 to 110%. Limits of detection for SPE-LC ES-MS were in the range of low ng l(-1), which demonstrates the suitability of the method for environmental samples. PMID- 11873960 TI - Retention characteristics of an immobilized artificial membrane column in reversed-phase liquid chromatography. AB - Retention for a varied group of compounds on an immobilized artificial membrane column (IAM PC DD2) with a methanol-water mobile phase is shown to fit a second order model for the retention factor (log k) as a function of the volume fraction of organic solvent. The numerical value of the intercept obtained by linear extrapolation to zero organic solvent (log k(w)) is shown to depend on the range of mobile phase composition used for the extrapolation. Each series of intercepts so obtained represents a different hypothetical distribution system as identified by the system constants of the solvation parameter model. Although a linear model is a poor fit for isocratic retention data, the linear solvent strength gradient model provides a reasonable estimate of isocratic retention factor values that are (slightly) larger than experimental values, but provide the same chemical information for the system. These preliminary results suggest that gradient elution may prove to be a rapid and useful method for creating system maps for column characterization and method development. In this work a system map is provided for methanol-water compositions from 0 to 60% (v/v) methanol and additional system constants for acetonitrile-water compositions containing 20 and 30% (v/v) acetonitrile. It is shown that the main factors contributing to retention on the IAM PC DD2 column are favorable cavity formation and dispersion interactions, electron lone pair interactions and the hydrogen-bond basicity of the sorbent. The latter feature more than any other distinguishes the IAM column from conventional chemically bonded phases. Interactions of a dipole-type (weakly) and inability to compete with the mobile phase as a hydrogen-bond acid reduce retention. A comparison of system constant ratios is used to demonstrate that the retention properties of the IAM column are not easily duplicated by conventional chemically bonded phases. The retention characteristics of the IAM column, however, are strongly correlated with the retention properties of pseudostationary phases used for micellar electrokinetic chromatography, which provide a suitable alternative to IAM columns for physical property estimations. By the same comparative method it is shown that retention on the IAM column possesses some similarity to biomembrane absorption processes, allowing suitable correlation models to be developed for the estimation of certain biopartitioning properties. PMID- 11873961 TI - Characterization and origin identification of 2,4,6-trinitrotoluene through its by-product isomers by liquid chromatography-atmospheric pressure chemical ionization mass spectrometry. AB - The by-products of industrial 2,4,6-trinitrotoluene (TNT), including isomers of trinitrotoluene, dinitrotoluene, trinitrobenzene and dinitrobenzene were investigated using liquid chromatography-mass spectrometry (LC-MS), in order to build a profile for the characterization of TNT samples from various origins. LC MS with atmospheric pressure chemical ionization, in the negative-ion mode, was found to be the most suitable method for this study. The characterization of TNT by the by-product profile was demonstrated on a variety of TNT samples. PMID- 11873962 TI - Fast microwave-assisted dansylation of N-nitrosamines. Analysis by high performance liquid chromatography with fluorescence detection. AB - A fast microwave-assisted dansylation procedure has been developed for the derivatization of N-nitrosamines prior to high-performance liquid chromatography determination. N-Nitrosomorpholine, N-nitrosodimethylamine, N nitrosodiethylamine, N-nitrosopyrrolidine and N-nitrosopiperidine are first denitrosated by hydrobromic acid-acetic acid to produce secondary amines, which are then quantitatively dansylated in 5 min using radiation power of 378 W and a maximum pressure of 1.4 bar inside the reactor. The reaction mixture is separated on a C18 column with acetonitrile-water (55:45, v/v) as mobile phase with fluorimetric detection at 531 nm (excitation at 339 nm). The detection limits range from 8 to 75 pg for N-nitrosomorpholine and N-nitrosodiethylamine, respectively. The method was applied to study the recoveries of N-nitrosamines in beer and their determination in cigarette smoke. PMID- 11873963 TI - Cutinase-peptide fusions in thermoseparating aqueous two-phase systems. Prediction of partitioning and enhanced tag efficiency by detergent addition. AB - It is of increasing importance to develop efficient purification methods for recombinant proteins where the number of steps can be minimised. The aim has been to establish a method for predicting the partitioning of the wild-type target protein in an aqueous two-phase system, and with this as basis, develop fusion tags and optimise the phase system for enhanced partitioning of the target protein. The surface of the lipolytic enzyme cutinase from Fusarium solani pisi was investigated with a computer program, Graphical Representation and Analysis of Surface Properties (GRASP). The accessible surface areas for the different amino acid residues were used together with peptide partitioning data to calculate the partition coefficient for the protein. The separation system was composed of a thermoseparating random copolymer of ethylene oxide and propylene oxide. Breox PAG 50A 1000, as top phase forming polymer and a hydroxypropyl starch polymer, Reppal PES 200, as bottom phase polymer. The calculated partition coefficient for the wild-type protein (K= 1.0) agreed reasonably well with the experimentally determined value (K=0.85). Genetic engineering was used to construct fusion proteins expressed in Saccharomyces cerevisiae based on cutinase and peptide tags containing tryptophan, to enhance the partitioning in aqueous two-phase systems. The partitioning of the cutinase constructs could qualitatively be predicted from peptide partitioning data, i.e. the trends in partitioning could be predicted. A spacer peptide introduced between protein and tag increased the partitioning of the protein towards the ethylene oxide propylene oxide (EOPO) copolymer top phase. The aqueous two-phase system was modified by addition of detergent to increase the partitioning of the cutinase variants towards the EOPO copolymer phase. Triton and a series of C12En detergents selectively increased the partitioning of cutinase constructs with (WP)4-based tags up to 14 times compared to wild-type cutinase. The protein partition could almost quantitatively be predicted from the peptide partition data. PMID- 11873964 TI - Purification of Food Color Red No. 106 (acid red) using pH-zone-refining counter current chromatography. AB - pH-Zone-refining counter-current chromatography was successfully applied to the separation of the main components of Food Color Red No. 106 (R-106, acid red, Color Index No. 45100). A 300-mg quantity of sample was separated using the following two-phase solvent system: n-butanol-water, 40 mM sulfuric acid in organic stationary phase and 30 mM ammonia in aqueous mobile phase. The obtained fractions were analyzed by high-performance liquid chromatography and fast atom bombardment mass spectrometry. The separation yielded 261.9 mg of main component of acid red with purity of 99.9%. PMID- 11873965 TI - Determination of triton X-100 in plasma-derived coagulation factor VIII and factor IX products by reversed-phase high-performance liquid chromatography. AB - Plasma protein pools are often virus-inactivated by the solvent-detergent method, using tri-n-butyl phosphate and Triton X-100, followed by removal and determination of these compounds. We used reversed-phase high-performance liquid chromatography for the determination of Triton X-100 in coagulation factor VIII and factor IX products, Octonativ-M and Nanotiv, respectively (Pharmacia, Stockholm, Sweden). The chromatographic system included a C18 silica column and a linear acetonitrile gradient. The advantage of this method is the low detection limit (0.3 microg/ml) combined with detection at 280 nm, which gives a more stable baseline and has less interference from other compounds. As compared to other methods, where shorter wavelengths are used. PMID- 11873966 TI - Dynamic coating ion-exchange chromatography of cations on an octadecyl-bonded silica stationary phase. AB - It was found that common cations (Na+, NH4+, K+, Mg2+ and Ca2+) could be strongly retained on an ODS stationary phase when aqueous solutions of carboxylic acids were used as eluents. The chromatographic conditions used in this work were the same as in common cation-exchange chromatography on a cation-exchange resin and the retention behavior of the above-mentioned cations on the ODS column was quite similar to that on a cation-exchange column. The retention behavior and mechanism have been investigated using a number of carboxylic acids as eluents. The retention mechanism of the cations in these experiments was considered to be a dynamic coating ion-exchange mechanism. The carboxylic acids in the mobile phase were coated onto the surface of the ODS stationary phase and formed a dynamic carboxylic acid functional layer which could act like the functional group layer of a carboxylic group cation exchanger. PMID- 11873967 TI - Multiple headspace extraction-gas chromatographic method for the study of process kinetics. AB - A multiple headspace extraction gas chromatographic technique to automate the study of process kinetics that involves volatile species was developed in this study. The technique was demonstrated by using it to study methanol formation in kraft black liquor samples under an isothermal temperature of 70 degrees C. The results indicate that methanol formation in kraft black liquors under isothermal conditions follows an exponential decay function. The present method is very simple, efficient and fully automated. It can be easily applied to study slow kinetic processes, such as reaction or adsorption and desorption, involving volatile species in any environmental and industrial samples with complicated matrices. PMID- 11873968 TI - Characterization of phosphorus-containing gas chromatographic stationary phases by linear solvation energy relationships. AB - Linear solvation energy relationships allow the prediction of a variety of solubility interactions based on a set of descriptors found in the following equation: [equation: see text]. SP refers to an intrinsic thermodynamic property that can be found experimentally for a series of solutes. Phases containing phosphate, phosphite and phosphine functional groups were studied in this work. Coefficients obtained during this work, as well as those available for previously characterized phases, were correlated with molecular structural descriptors. When effects of non-phosphorus functional groups are estimated and subtracted out, hydrogen bond acceptor capability, a1, shows a positive trend when correlated with percent functional group. Correlation of the dipolarity/polarizability coefficient, s, with calculated atomic polarizability shows stationary phases group according to like functional groups. A similar correlation with dipole moment gives a trend of increasing dipole as s1 increases. Further quantitative structure-solubility relationship work is planned to better describe the contributions of inner shell and valence electrons to the chemical and physical properties of these compounds. PMID- 11873969 TI - Separation of racemic sulfoxides and sulfinate esters on four derivatized cyclodextrin chiral stationary phases using capillary gas chromatography. AB - The separation of 17 chiral sulfoxides and eight chiral sulfinate esters by gas chromatography (GC) on four derivatized cyclodextrin chiral stationary phases (CSPs) (Chiraldex G-TA, G-BP, G-PN, B-DM) is presented. Many of these compounds are structural isomers or part of a homologous series. Differences in enantioselectivity of the methyl phenyl sulfoxide isomers on the derivatized gamma cyclodextrin and the heptakis 2,6-di-O-methyl-beta-cyclodextrin (i.e. B-DM) CSPs are discussed. Under the conditions of this study, the molecular mass cut off for the GC separation of these compounds was approximately 230. Compounds of higher molecular mass were not eluted from the CSPs at reasonable times and temperatures, but these higher molecular mass enantiomers can be separated by liquid chromatography and capillary electrophoresis. The enantiomeric separation and elution order of a sulfinate ester containing two stereogenic centers as well as 15 chiral sulfoxides is presented. The G-TA and B-DM CSPs generally gave opposite elution orders for most of the compounds studied. PMID- 11873971 TI - Isolation and identification of diisopropylnaphthalene isomers in the alkylation products of naphthalene. AB - GC analysis of diisopropylnaphthalene (DIPN) isomers is presented. Naphthalene was repeatedly alkylated with propylene over an amorphous aluminosilicate catalyst under different preparation conditions, and then pure isomers or at least DIPN fractions enriched in individual isomers were isolated from the alkylates and analysed using GC-MS, IR and NMR techniques. Eight of the ten possible isomers appeared in the chromatograms; seven of them (1,3-, 1,4-, 1,5-, 1,6-, 1,7-, 2,6- and 2,7-) were separated and identified, while the eighth one was indirectly proved to be 2,3-DIPN. The remaining two isomers (1,2- and 1,8 DIPN) were not found in the products. PMID- 11873970 TI - Determination of polychlorinated biphenyls in biota samples using simultaneous pressurized liquid extraction and purification. AB - In order to reduce time and cost of analysis, a new pressurised liquid extraction method that automatically and rapidly achieves quantitative and selective (i.e., lipid-free) extraction of polychlorinated biphenyls (PCBs) in biota tissues was optimized. It consists of on-line clean-up by inclusion of sorbents in the extraction cell. The freeze-dried sample is dispersed with Florisil and loaded in the extraction cell containing an extra amount of Florisil. The extraction is performed under mild conditions using 55 ml of a dichloromethane-pentane (15:85) mixture, a temperature of 40 degrees C, a static extraction time of 10 min and two extraction cycles. The Florisil retains coextracted lipids from the matrix, and the extract, after pre-concentration, is clean enough for direct injection into GC-MS and GC-electron-capture detection (ECD). Quantitative recoveries (from 90 to 106%) are obtained for both native and spiked PCB congeners in samples with a high lipidic content (up to 42% dry mass, in spoonbill eggs). The reproducibility of replicate extractions was better than 11% relative standard deviation. Method detection limits were in the ranges of 0.001-0.004 and 0.002 0.07 ng g(-1) dry mass for GC-ECD and GC-MS-MS, respectively. The method was validated using the standard reference material SRM 2974 (a mussel tissue) from the US National Institute of Standards and Technology, compared to Soxhlet and matrix solid-phase dispersion extraction methods, and used to evaluate the contamination by PCBs in bivalves from South of Spain. PMID- 11873972 TI - Analysis of a stable halogenated derivative of muramic acid by gas chromatography negative ion chemical ionization tandem mass spectrometry. AB - Muramic acid (Mur) is present in the cell wall of Eubacteria and serves as a chemical marker for the trace detection of bacteria and bacterial cell wall debris in complex matrices. There have been numerous studies using a variety of derivatives of Mur, particularly in combination with gas chromatography-tandem mass spectrometry (GC-MS-MS) where the detection limit has been steadily lowered. A stable, halogenated derivative, the pentafluorobenzyl oxime (PFBO) acetate of Mur, has been developed by others and successfully used for GC with electron capture detection. The current report is the first use of this derivative for GC MS-MS analysis of Mur, or indeed any other carbohydrate, using negative ion chemical ionization (NICI) with GC-MS-MS. Mur was readily detected in settled surface dust (166 ng/mg), as well as dust collected from indoor air (1.4-5.9 ng/mg). Analyses of Mur as a PFBO acetate by GC-NICI-MS-MS or as alditol acetates by electron impact GC-electron impact ionization MS-MS serve as complementary approaches for trace detection in complex matrices. PMID- 11873973 TI - Determination of oxadiazon residues by headspace solid-phase microextraction and gas chromatography-mass spectrometry. AB - A method for the determination of trace amounts of the herbicide oxadiazon was developed using headspace solid-phase microextraction (HS-SPME), gas chromatography-mass spectrometry (GC-MS) and selected ion monitoring. It was applied to determine oxadiazon in ground water, agricultural soil, must, wine and human urine samples. To determine oxadiazon in liquid samples, a response surface methodology generated with a Doehlert design was applied to optimize the HS-SPME conditions using a 100 microm polydimethylsiloxane fibre. For the analysis of soil samples, they were mixed with water and the SPME fibre suspended in the headspace above the slurry. Ground water, human urine and must show linear concentration range of application of 0.5-50 ng ml(-1)' with detection limits < or =0.02 ng ml(-1). HS-SPME-GC-MS analysis yielded good reproducibility (RSD values between 6.5 and 13.5%). The method validation was completed with spiked matrix samples. The developed analytical procedure is solvent free, cost effective and fast. PMID- 11873974 TI - Selective fluorometric detection of polyamines using micellar electrokinetic chromatography with laser-induced fluorescence detection. AB - The polyamines putrescine, cadaverine, spermine and spermidine were separated and quantified by micellar electrokinetic chromatography (MEKC) with laser-induced fluorescence detection. The derivatization reagent, 1-pyrenebutanoic acid succinimidyl ester (PSE), allowed for the selective detection of the polyamines at 490 nm. Multiple labeling of the polyamines with PSE allows the formation of intramolecular excimers that emit at longer wavelengths (450-520 nm) than mono labeled analytes (360-420 nm). Optimal separation of the labeled polyamines was achieved using a separation buffer consisting of 10 mM phosphate pH 7.2, 30 mM cholate, and 30% acetonitrile. Using these conditions, the four polyamines were separated in under 10 min. Limits of detection for putrescine, cadaverine, spermine and spermidine were 6, 5, 15 and 13 nM, respectively. These are superior or comparable to those previously reported in the literature using fluorescence detection. PMID- 11873975 TI - Solid-phase extraction and sample stacking-capillary electrophoresis for the determination of quaternary ammonium herbicides in drinking water. AB - Conditions for the simultaneous determination of paraquat, diquat and difenzoquat by capillary zone electrophoresis were established by combining two preconcentration procedures. Off-line solid-phase extraction was used for the isolation and preconcentration of quats in drinking water. Quats were then analysed by capillary electrophoresis using sample stacking with matrix removal as on-column preconcentration procedure. Two different porous graphitic carbon cartridges were compared. The breakthrough volumes of the three herbicides were calculated and the loading capacity of the sorbents was compared. Recoveries higher than 80% for difenzoquat and around 40% for paraquat and diquat were obtained when a sample volume of 250 ml was percolated. For the stacking capillary electrophoresis analysis of quats, 50 mM acetic acid-ammonium acetate (pH 4.0), 0.8 mM cetyltrimethylammonium bromide with 5% (v/v) methanol as carrier electrolyte was used. Detection limits, based on a signal-to-noise ratio of 3:1, were lower than 0.3 microg l(-1) for standards in Milli-Q water, and lower than 2.2 microg l(-1) for drinking water samples. Run-to-run and day-to-day precision of the method were established. The two preconcentration procedures used together was successfully applied to the analysis of the three herbicides in spiked drinking water at concentrations below the maximum admissible US Environmental Protection Agency levels. PMID- 11873976 TI - Polyallylamine-grafted cellulose gel as high-capacity anion-exchanger. AB - A new cellulose-based anion-exchanger was prepared by grafting polyallylamine onto cellulose. The material was obtained by partial oxidation of a size exclusion grade cellulose gel by aq. NaIO4, forming dialdehyde cellulose, followed by Schiff base formation with a polyallylamine (PAA, molecular mass 5000) and subsequent reduction for stabilization. Three grades of PAA-cellulose gels, with amino group contents of 0.78, 1.01 and 1.28 mmol/g cellulose, were examined for their ionic interaction with mono- and divalent carboxylic acids at pH 2.5-5.5. While the retention factor for monovalent acids was nearly proportional to the amino group content of the gel, that for divalent acids was remarkably greater for the PAA-cellulose gel than for the conventional diethylaminoethyl (DEAE) cellulose gel bearing more amino groups (1.97 mmol/g cellulose). Such high capacity can be explained by the high local density of amino groups on grafted PAA, in contrast to the random and sparse charge distribution in conventional exchangers. PMID- 11873977 TI - Determination of phenolic xenoestrogens in water by liquid chromatography with coulometric-array detection. AB - A sensitive method for the simultaneous determination of phenolic xenoestrogens such as bisphenol A, 2,4-dichlorophenol, 4-tert,-butylphenol, 4-n2-pentylphenol, 4-n-hexylphenol, 4-n-heptylphenol, 4-octylphenol, 4-nonylphenol was developed using reversed-phase LC and coulometric-array detection. Stepwise gradient elution with phosphoric acid in water-acetonitrile was used. The calibration curves were linear in the range of 5.0 (or 10.0)-1000 ng ml(-1) with correlation coefficients of 0.9978-0.9999, the limits of detection were 0.01-0.02 ng ml(-1). Sample clean-up was performed by solid-phase extraction (SPE) using 3M Empore extraction disks. Three commercial sorbents, C18, SDB-XD (styrene-divinylbenzene polymer) and SDB-RPS (sulfonated styrene-divinylbenzene polymer) were compared. The highest recoveries were obtained with SDB-RPS. They were above 70% with a relative standard deviation of less than 15%. The proposed method was applied to the determination of phenolic xenoestrogens in various water samples. PMID- 11873978 TI - Development and validation of a reversed-phase high-performance liquid chromatographic method for the determination of ethyl-3-(N-n-butyl-N acetyl)aminopropionate in an insect repellent semi-solid formulation. AB - A reversed-phase high-performance liquid chromatographic method with detection at 220 nm was developed and validated for the determination of ethyl-3-(N-n-butyl-N acetyl)aminopropionate, IR 3535, in an insect repellent semi-solid product. A Hypersil ODS RP-C18 column (250 x 4.6 mm), 5 microm particle size, was equilibrated with a mobile phase consisted of water-acetonitrile (60:40, v/v). Its flow-rate was 1.0 ml/min. Excipients did not interfere with the determination of IR 3535 (Rs = 8.663). Intra- and inter-day relative standard deviations for samples were not higher than 0.61 and 1.2%, respectively. Mean recovery was found not lower than 98.5% and not higher than 100.3%. The method of external standard was adopted. Calibration curves were linear in the concentration range between 1.0 x 10(-6) and 5.0 x 10(-4) M. Limits of detection and quantitation were 65 and 196 ng/ml, respectively. PMID- 11873979 TI - Observation of sorptive losses of volatile sulfur compounds during natural gas sampling. AB - The reason for the compound-dependent over-estimation of the recoveries of several volatile organic sulfur compounds when using a Silcosteel cylinder for sample storage as reported earlier was examined. From the different possible sources of errors that were taken into consideration, the silicone tubing, which was used to fill a standard Tedlar sample bag for calibration, was identified as the cause of the artefact. The comparison of different tubing materials showed that PTFE is the best choice since it causes only minor losses (<10%) of propyl- and butylmercaptans. PMID- 11873980 TI - Effect of the organic modifier concentration on the retention in reversed-phase liquid chromatography II. Tests using various simplified models. AB - Ten simplified expressions for the retention factor, k', that arise from either the adsorption or partition mechanism for retention in reversed-phase chromatographic columns are examined in what concerns the model they express and their performance to fit experimental data. In order to test the simplified expressions, which describe the variation of the retention of a solute with the organic modifier content in the mobile phase, a wide range of solutes in mobile phases modified with three different organic modifiers was used. It is shown that a new three-parameter expression of ln k' works more satisfactorily, since it combines simplicity, high applicability and good numerical behavior. It is also shown that the applicability of a simplified equation does not entail the validity of its model and thus no molecular information can be gained from its use. PMID- 11873981 TI - Mechanistic implications of the equality of compensation temperatures in chromatography. AB - A common interpretation of the observation that two processes exhibit similar compensation temperatures in an enthalpy-entropy plot is that the two processes occur via the same "mechanism". We show that this interpretation is not rigorously allowed. In fact, the only thing that can be concluded from the observation of identical compensation temperatures is that the relative contributions of enthalpy and entropy to the overall free energy are the same in the two processes. Since it is possible that two processes occur via different mechanisms that, by chance, result in the same relative blends of enthalpy and entropy, the observation of identical compensation temperatures cannot be used as evidence for mechanistic identity. If two processes exhibit different compensation temperatures, however, it can logically be concluded that the two processes are mechanistically distinct. PMID- 11873982 TI - Chromatographic explanation for the side-wall induced band broadening in pressure driven and shear-driven flows through channels with a high aspect-ratio rectangular cross-section. AB - Based on a simple, chromatography-based analogy, a quantitatively exact explanation for the strong additional band broadening induced by the presence of the side-walls in flow channels with a large aspect-ratio rectangular cross section is given and validated for two different flow types: pressure-driven and shear-driven flow. PMID- 11873983 TI - Evaluation of capillary liquid chromatography-electrospray ionization ion mobility spectrometry with mass spectrometry detection. AB - Due to the proteomics revolution, multi-dimensional separation and detection instruments are required to evaluate many peptides and proteins in single samples. In this study, electrospray ionization (ESI) ion mobility spectrometry (IMS) was evaluated as an additional separation after HPLC separations. Common HPLC mobile phase compositions (solvents, acid modifiers, and buffers) were assessed for the effect on ESI-IMS response. Up to 5 mM sodium phosphate, a non volatile buffer, was able to be electrosprayed into the IMS without degradation of the instrumental performance. Due to the rapid separation times of IMS, multiple IMS spectra were obtained within a single HPLC peak. A five-peptide mixture was separated in a capillary HPLC column under isocratic conditions within 3 min. Coelution of two peaks due to non-optimal HPLC conditions occurred and these two peaks could not be distinguished by HPLC with UV detection. In contrast, the single ion mobility chromatograms provided separation of each peptide as well as providing a second degree of analyte identification (HPLC retention time and IMS mobility). Furthermore, IMS-MS analysis of the five peptides and comparison with HPLC retention times showed that each peptide had a unique retention time-ion mobility-mass to charge value. This work showed that IMS could be employed for direct separation and detection of HPLC eluents and also could be combined with HPLC-MS for three unique dimensions of separation. PMID- 11873984 TI - Molecular mass distribution analysis of ethyl(hydroxyethyl)cellulose by size exclusion chromatography with dual light-scattering and refractometric detection. AB - Dual low-angle light scattering and refractometric detection coupled to size exclusion chromatography provided proof for the presence of a low amount of stable aggregates/particles in ethyl(hydroxyethyl)cellulose. Unlike the correct size-exclusion chromatographic behavior of the parent polysaccharide itself, the aggregates exhibit variable size-dependent weak retention as a function of flow rate and of ionic strength of the aqueous mobile phase. Therefore, determination of the molecular mass of non-aggregated polymer is possible in aqueous mobile phase containing 0.1 M NaCl under conditions at which aggregates are completely adsorbed on the column packing irrespective of the flow-rate used. Flow-rate and ionic strength-dependent variations of aggregate behavior as well as model size exclusion experiments with latex particles indicate that they partly carry a minute charge and have a compact structure. Their weak retention under the separation conditions used suggests a difference in their surface chemistry when compared with the dissolved polymer coils which exhibit a correct size-exclusion behavior. PMID- 11873985 TI - Enhancement of selectivity in reversed-phase liquid chromatography. AB - In an effort to gain insight into the relationship between stationary phase solvation and selectivity, the use of short- and medium-chained-length alcohols (methanol, n-propanol, n-butanol, and n-pentanol) as mobile phase modifiers in reversed-phase liquid chromatography (RPLC) was investigated to determine their impact on chromatographic selectivity. A wide range of mobile phase compositions was evaluated because of the large effect exerted by solvent strength on selectivity. Employing a set of six vanillin compounds as retention probes, evidence is presented to support the view that an increase in the hydrophobicity of the organic modifier used in RPLC can increase the selectivity of the C18 alkyl bonded phase while simultaneously decreasing the retention time of the eluting solutes. Thus, we are presented with an interesting paradox: higher selectivity and shorter retention times, which can be attributed to changes in either solvent selectivity and/or stationary phase solvation by the organic modifier. PMID- 11873986 TI - Effect of the organic modifier concentration on the retention in reversed-phase liquid chromatography I. General semi-thermodynamic treatment for adsorption and partition mechanisms. AB - A semi-thermodynamic treatment is adopted to account for adsorption or partition of solute molecules from aqueous mobile phases on/in reversed-phase liquid chromatography stationary phases. The theoretical expressions of ln k' versus organic modifier content are tested against 10 data sets covering a variety of solute molecules. It is shown that the mean field approximation, adopted widely in ptevious studies, is marginally valid in aqueous mobile phases, especially in the presence of solute molecules, and the lattice model approximation, which is also used in relevant studies, is a poor approximation. Clear conclusions about the validity of either the adsorption or the partition model for the retention mechanism could not be drawn. The equations of the adsorption model describe all data sets absolutely satisfactorily and yield a physically reasonable picture about the behavior of modifier and solvent at the adsorbed layer. However, the high applicability of the adsorption model may not safely entail the validity of the adsorption mechanism at a molecular level, especially in the case of solutes with small and non-polar molecules, where our analysis gives strong indications about the validity of the partition mechanism. The next steps needed for the final elucidation of the retention mechanism in reversed-phase chromatographic columns are indicated. PMID- 11873987 TI - Sulfoacylated poly(styrene-divinylbenzene) copolymers as resins for cation chromatography. Comparison with sulfonated, dynamically coated and silica gel cation exchangers. AB - Most important properties of an ion chromatographic resin are influenced by the resin matrix, the type of functional group and the ion-exchange capacity. Highly crosslinked PS-DVB resins of 5 microm diameter have been functionalized by sulfoacylation, by sulfonation and by dynamic coating over a wide range of exchange capacities. These materials allow a study of the influence of different ion-exchange sites and capacities. The influence of the degree of functionalization on resin performance is completely reverse for sulfoacylated and sulfonated resins. The HETP values for all observed analytes (Cu, Pb, Zn, Ni, Co, Cd, Mn, Ca, Mg) in a tartaric acid elution system decrease for sulfoacylated resins with increasing capacity, for sulfonated resins with decreasing capacity. The performance of sulfoacylated exchangers is better than for dynamically coated ones and far better than for sulfonated resins. The performance of silica gel based cation-exchangers such as BioSil CAT is in most cases better than observed for sulfoacylated resins. PMID- 11873988 TI - Methacrylate monolithic columns of 320 microm I.D. for capillary liquid chromatography. AB - Monolithic capillary columns (320 microm I.D.) were prepared for capillary liquid chromatography (CLC) by radical polymerization of butylmethacrylate (BMA) and ethylenedimethacrylate (EDMA) in the presence of a porogen solvent containing propan-1-ol, butane-1,4-diol and water. The influence of the contents of the porogen solvent and EDMA in the polymerization mixture on the monolith porosity and column efficiency was investigated. The composition of the polymerization mixture was optimized to attain a minimum HETP of the order of tens of microm for test compounds with various polarities. The separation performance and selectivity of the most efficient monolithic column prepared was characterized by van Deemter curves, peak asymmetry factors and Walters hydrophobicity and silanol indices. It was demonstrated that the 320-microm I.D. monolithic column exhibited CLC separation performance similar to that observed for 100- and 150-microm I.D. monolithic columns reported in the literature; moreover, the 320-microm I.D. column was easier to operate in CLC and exhibited a higher sample loadability. PMID- 11873989 TI - Collection in analytical-scale supercritical fluid extraction. AB - This review is a comprehensive summary of available collection techniques in supercritical fluid extraction (SFE), with emphasis on which parameters are especially important for a successful analyte collection. Environmental, biological and agricultural applications, including several types of sample matrices and analyte groups, are discussed with respect to choice of collection mode and optimization of collection conditions. This review also includes discussions about collection when a modifier is used or when the sample contains large amounts of fat or water, as well as possibilities to achieve enhanced selectivity. PMID- 11873990 TI - Gas chromatographic-mass spectrometric characterization of all acyclic C5-C7 alkenes from fluid catalytic cracked gasoline using polydimethylsiloxane and squalane stationary phases. AB - Published retention indices of acyclic alkenes C5-C7 on squalane and polydimethylsiloxane as stationary phases were investigated, and reliable retention indices of alkenes from various sources were converted to separation systems used in a laboratory. Retention indices measured on available authentic commercial alkenes and on alkenic fraction of gasoline, published retention indices as well as means of GC-MS were used for verification of calculated retention indices. Retention of some gas chromatographic unseparated isomer pairs was obtained by mass spectrometric deconvolution using a specific single-ion monitoring. On the basis of these retention data, C5-C7 alkenes were identified and analyzed in the gasoline from fluid catalytic cracking. In the gasoline all 59 acyclic C5-C7 isomeric alkenes were determined at significantly different concentration levels. PMID- 11873991 TI - Analysis of endosulfan isomers and endosulfan sulfate in air and tomato leaves by gas chromatography with electron-capture detection and confirmation by gas chromatography-mass spectrometry. AB - Rapid analytical methods for the determination of endosulfan isomers and endosulfan-sulfate in air and plant samples were developed. The insecticides were trapped from air using a column containing Florisil and extracted with a low volume of ethyl acetate, assisted by sonication. Pesticide residues were determined by gas chromatography with electron-capture detection using a nonpolar capillary column. Residue identities were confirmed by gas chromatography coupled with mass spectrometry. Recoveries of these compounds from air samples were always higher than 78% with an RSD lower than 11% and the detection limits obtained were at least 0.3 ng/l air. Leaf samples were homogenised with ethyl acetate and extracts cleaned-up on an aluminium oxide column. Pesticides were eluted with a hexane-ethyl acetate (80:20, v/v) mixture. Recoveries obtained from plant samples were higher than 78% with an RSD lower than 14% and detection limits in leaves were 0.02 microg/g for each pesticide. These methods were applied to study the volatilisation of endosulfan from tomato leaves under laboratory conditions. A volatilisation rate near 1% of the initial amount of endosulfan per hour was obtained during the first 24 h at room temperature. PMID- 11873993 TI - Rational approach to quantitative sodium dodecyl sulfate capillary gel electrophoresis of monoclonal antibodies. AB - Sodium dodecyl sulfate capillary gel electrophoresis has been used to separate and quantify murine monoclonal antibodies. The method uses a murine IgG, whose subclass differs from the analyte antibody, as an internal reference. The internal reference is chosen based on knowing that mouse IgG1 can be separated from mouse IgG2a or IgG2b. Good intra- and inter-day reproducibility [relative standard deviation (RSD)<2%] of peak-area ratio has been obtained. A calibration curve also demonstrates high linearity (R2=0.9999) of response for the analyte. The described method is highly suitable for accurate determination of the antibody concentration even if a capillary electrophoresis apparatus is unable to provide good injection reproducibility. PMID- 11873992 TI - Novel approach for the simultaneous analysis of glyphosate and its metabolites. AB - A novel approach for the simultaneous analysis of glyphosate (PMG), and aminomethylphosphonic (AMPA, GlyP), N-methylaminomethylphosphonic (MAMPA. SarP) and methylphosphonic (MPA) acids is presented. This includes a preliminary 31P NMR analysis of mixtures of PMG, MPA, AMPA and MAMPA, their further derivatization to volatile phosphonates by means of the trifluoroacetic acid trifluoroacetic anhydride-trimethyl orthoacetate reagent and subsequent MS [chemical ionization (CI) MS, GC-CI-MS, GC-electron impact ionization MS] and/or GC-flame ionization detection (FID) analysis of the products of derivatization. The detection limits of PMG, AMPA, MAMPA and MPA by means of GC-CI-MS and GC-FID were determined. The calibration graphs (GC-FID) for these derivatives were in the range 0.1 to 100 nmol linear and sufficiently reproducible for quantitative determinations. The applicability of the method was demonstrated during the analysis of water samples fortified with PMG, AMPA and MAMPA, characterized by recoveries of >95%. PMID- 11873994 TI - Impurity profiling of ephedrines in methamphetamine by high-performance liquid chromatography. AB - Separation of the enantiomers and diastereomers of ephedrines was investigated for impurity profiling of methamphetamine. We describe a method for the analysis of (1S,2R)-(+)-ephedrine, (1R,2S)-(-)-ephedrine, (1S,2S)-(+)-pseudoephedrine and racemic methylephedrine in bulk methamphetamine by HPLC using two different columns: a phenyl-beta-cyclodextrin-type column and an ODS-type column. The analytes were detected by UV absorbance measurement at 210 nm. As little as 0.05% of each ephedrine in bulk methamphetamine could be determined. In the impurity profiling of methamphetamine, the identification of ephedrines may provide valuable information about the precursor. This method was confirmed to be sufficiently sensitive to identify trace amounts of (1R,2S)-(-)-ephedrine and (1S,2S)-(+)-pseudoephedrine in bulk methamphetamine synthesized by the Emde method. PMID- 11873995 TI - Supercritical carbon dioxide extraction of volatiles from spices. Comparison with simultaneous distillation-extraction. AB - Supercritical fluid extraction (SFE) and simultaneous distillation-extraction (SDE) were used to extract the essential oils from three different spices (oregano, basil, and mint), and a comparative study of extracts obtained using SFE and SDE is presented. Temperature and pressure for the SFE extraction were optimized prior to the experimental extractions. The extracts obtained using the two methods were very similar in composition, but SFE yielded better relative standard deviations and avoided the thermal degradation or solvent contamination of samples. PMID- 11873996 TI - Micellar liquid chromatography: suitable technique for screening analysis. AB - The screening capability of micellar liquid chromatography (MLC) is discussed using the reported chromatographic data of several sets of compounds (amino acids, beta-blockers, diuretics, phenethylamines, phenols, polynuclear aromatic hydrocarbons, steroids and sulfonamides) and new results (sulfonamides and steroids). The chromatographic data are treated with an interpretive optimisation resolution procedure to obtain the best separation conditions. Usually, the pH and the concentration of surfactant (sodium dodecyl sulfate, SDS, or cetyltrimethylammonium bromide) for the optimal mobile phase were 2.5-3 and < 0.12 M, respectively. The nature and concentration of organic solvent depended on the polarity of the eluted compounds: a low volume fraction of propanol (approximately 1%, v/v) was useful to separate the amino acids, with log P(o/w) < -1 (where P(o/w) is the octanol-water partition coefficient). A greater concentration of this solvent (approximately 5-7%) was needed for compounds in the range -1 < log P(o/w) < 2, as with the studied diuretics and sulfonamides, and a high concentration of propanol (approximately 15%) or a low concentration of butanol (< 10%) had to be used for less polar compounds with 1 < log P(o/w) < 3, such as the beta-blockers. Pentanol (< 6%) was more suitable for the even less polar compounds with log P(o/w) > 3, such as the steroids. For basic drugs such as the phenethylamines (0 < log P(o/w) < 1.7), eluted with a micellar eluent of anionic SDS, propanol was too weak. A study is also shown for mixtures of sulfonamides (log P(o/w) = -1.2 to 1.7) and steroids (log P(o/w) = 3.0-8.1) eluted from conventional C18 columns with SDS mobile phases containing acetonitrile and 1-pentanol, respectively, which are compared with classical acetonitrile-water and methanol-water mixtures. The results complement a previous study on beta-blockers (log P(o/w) = -0.03 to 2.8) and reveal that MLC is a very competitive technique for the screening of compounds against conventional RPLC, due to its peculiar behaviour with regard to the selectivity and elution strength. The concentration of organic solvent needed to obtain sufficiently low retention times (even for highly hydrophobic steroids with log P(o/w) = 7-8) is also appreciably smaller for MLC, which reduces the environmental impact of the mobile phases. PMID- 11873997 TI - Retention of ionizable compounds in high-performance liquid chromatography. IX. Modelling retention in reversed-phase liquid chromatography as a function of pH and solvent composition with acetonitrile-water mobile phases. AB - The influence of pH and solvent composition of acetonitrile-water mobile phases on the retention of acids and bases on a polymeric stationary phase is studied. Very good relationships between retention and mobile phase pH are obtained if the pH is measured in the proper pH scale. The fit of retention to pH for a particular solvent composition provides the pKa values of the equilibria between the different acid-base species and the retention parameters of these species at this solvent composition. Several models are tested that relate these parameters to solvent composition and properties in order to propose a general model to predict retention for any mobile phase pH and composition. PMID- 11873998 TI - Application of the "VARICOL" process to the separation of the isomers of the SB 553261 racemate. AB - A new continuous chromatographic process (VARICOL) has been presented recently. The basic principle of the new VARICOL process consists of an asynchronous shift of the inlet/outlet lines in a multi-column system on a recycle loop. This process has been used to perform the separation of the optical isomers of the SB 553261 racemate. In this paper, we illustrate that for this specific separation, the VARICOL process is more efficient than the well-known SMB process. PMID- 11873999 TI - Screening approach for chiral separation of pharmaceuticals. Part I. Normal-phase liquid chromatography. AB - A strategy for rapid screening for the separation of chiral molecules of pharmaceutical interest by normal-phase liquid chromatography using three cellulose/amylose stationary phases is proposed. In a first step, the most important parameters for the separations were determined and studied for their effects by means of experimental designs. Results showed that the cellulose tris (3,5-dimethylphenylcarbamate), the amylose tris-(3,5-dimethylphenylcarbamate) and the cellulose tris-(4-methylbenzoate) stationary phases have very broad and complementary enantiorecognition properties. The type of organic modifier used in the mobile phase appeared to have a dramatic influence on the quality of the separation. Based on the results of the preliminary study, a screening strategy was developed and successfully applied to a set of 36 diverse drugs. Enantiomeric separation was observed in 89% of cases and the analysis times were usually shorter than 20 min. PMID- 11874000 TI - High-performance liquid chromatographic determination of furfural compounds in infant formulas. Changes during heat treatment and storage. AB - Furfural contents in adapted and follow-up infant formulas were measured by RP HPLC. The evolution of furfural compound contents during storage (a year at 20 and 37 degrees C) was studied. 2-Furylmethylketone and 5-methyl-2-furaldehyde were not detectable in analysed samples. The differences in the furfural compounds at point zero between both infant formulas has to be ascribed to the differences in protein and iron contents. An increase in free 5-hydroxymethyl-2 furfuraldehyde (HMF), 2-furaldehyde (F) and HMF+F contents was observed in all samples, although the differences were not statistically significant. The storage temperature affected the total HMF content and the storage time affected the total HMF and F contents. PMID- 11874001 TI - High-performance liquid chromatographic determination of tocopherols in infant formulas. AB - A method for the simultaneous determination of alpha-tocopherol acetate and alpha , delta-, and gamma-tocopherols by normal-phase high-performance liquid chromatography (HPLC) with a fluorescent detector in infant formula is proposed. The values obtained in the determination of the analytical parameters: linearity, precision, limit of detection and accuracy (analysis of a standard reference material, SRM 1846), confirm the quality of the method. The proposed method is useful for the determination of alpha-, delta-, and gamma-tocopherols and alpha tocopherol acetate in infant formulas at a low cost and in a total time of 2 h. PMID- 11874002 TI - Rehabilitative treatment after unsuccessful teeth replantation: a case report. AB - A nine-year-old boy suffered a severe facial trauma in a bicycle crash, which resulted in the avulsion and subluxation of upper central incisors. Teeth were repositioned and stabilized, attempting to achieve a successful replantation. Root resorption occurred, teeth were extracted, followed by orthodontic treatment, and esthetic procedures, modifying laterals and canines. This article reports an attempt and failure of replantation, providing alternatives for achieving patient's satisfaction, applying a variety of techniques and areas of dental profession. PMID- 11874003 TI - Interdisciplinary rehabilitation and prevention in a case with early and extensive loss of primary teeth. AB - The oral rehabilitation of a five-and-a-half year old boy is presented, who had an almost complete dental loss due to caries at the age of three years. Partial and complete dentures were provided to restore function and aesthetics, to prevent psychological and speech disorders and to reduce the severity of malocclusion and loss of space in the permanent dentition. The long-term treatment planning objectives are discussed in relation to orthodontic considerations, the establishment of the permanent dentition and the expectations for future growth. This case demonstrates the requirement for interdisciplinary involvement in the treatment planning for children with extensive tooth loss. PMID- 11874004 TI - Combining functional appliances in the straightwire system. AB - The Trainer for Braces (T4B) helps speed up fixed appliance therapy, by derotating teeth and pushing them into the line of the arch. It also aids treatment stability by reducing the influence of undesirable myo-functional habits and retraining the oral musculature. I issue a T4B to all my patients on the day of bracket placement. I have noticed a 30% reduction in treatment times for those patients who wear the T4B as directed. PMID- 11874005 TI - Pulse oximetry: a diagnostic instrument in pulpal vitality testing. AB - One hundred children with normal maxillary central and lateral incisors were subjected to vitality tests each by electrical and pulse oximetry. As a control population to confirm the readings, 10 known nonvital anterior teeth with complete endodontic fillings were tested. The systemic oxygen saturation values measured on the index finger of the patient served as the control for the comparison of the oxygen saturation values measure on the teeth. The SaO2 values obtained on the teeth were correlated with the electrical test readings. The correlation between the SaO2 readings and electrical testing readings were found to be negative i.e. as the values of the electrical pulp testing reading increased, the SaO2 values decreased. Since a reproducible SaO2 level is obtainable on the vital teeth, pulse oximetry has immediate clinical value in providing base line vitality data for the traumatised teeth. PMID- 11874006 TI - Effect of serial extraction alone on crowding: relationship between closure of residual extraction space and changes in dentition. AB - Mandibular dentitions from 33 subjects who had undergone serial extraction without appliances were analyzed at three stages: before extraction of deciduous canines (T1), after extraction of first premolars (T2), and at the end of the observation (T3). It was suggest that the mesial movement or tipping of the second premolars was associated with most of the space closure from T2 to T3, although the distal movement or tipping of canines might also contribute space closure. PMID- 11874007 TI - The morphology of the mandibular antegonial notches and facial symmetry. AB - Thirty randomly selected pre-treatment postero-anterior cranial radiographs of adolescent patients attending the orthodontic department, School of Dental Medicine, State University of New York at Buffalo comprised the sample in this study. The aims of this study were (1) to compare the depths of the right, and the left, mandibular antegonial notches, and (2) to determine whether the morphology of the antegonial notches bears a statistical relationship to some other transverse metrical characters of the face. The frontal cranial radiographs of thirty patients were digitized to determine the linear, and surface area, measurements of the right, and the left, antegonial notches as well as some transverse dimensions of the faces. An analysis of variance showed that no statistically significant difference existed between the measurements made by the two examiners, who digitized the radiographs. The data were analyzed by means of the Student's t-test. The results showed that there were statistically significant differences (P<0.05) between the measurements of the right, and the left, mandibular antegonial notches. The data also showed that there were highly statistically significant differences between the corresponding bilateral facial dimensions (P<0.001). The results of this study suggest that facial symmetry, as measured on a frontal skull radiograph, is associated with the respective depths of the right, and the left, mandibular antegonial notches. PMID- 11874008 TI - A comparative evaluation of oral midazolam with other sedatives as premedication in pediatric dentistry. AB - The purpose of present study was to evaluate the safety and efficacy of orally administered midazolam in children as a sedative agent and to compare it with two other older agents, triclofos and promethazine. The study was conducted on ninety child patients requiring some short dental procedure. All the patients were with a good physical status (ASA-I). The ages ranged between 3 and 9 years. The patients were randomized into three study groups: Group 1, midazolam, Group II, triclofos and Group III, promethazine, on the basis of the drugs to be administered. After administration of drugs in each group, the effects were evaluated in terms of onset of action, sedative effect, ease of treatment completion, recovery time and postoperative amnesia. Midazolam was found to be the best drug among the three to produce conscious sedation in children. PMID- 11874009 TI - Microleakage and penetration depth of three types of materials in fissure sealant: self-etching primer vs etching: an in vitro study. AB - Clinical preventive procedures must be done after a risk assessment. One of the risk factors is the occlusal morphology of the posterior teeth. These caries-free fissures must be sealed. This first in vitro experimentation of the study evaluated the microleakage and the penetration depth of three types of materials by Vivadent: Helioseal F, Tetric, Tetric Flow. The teeth were etched with phosphoric acid and bonded using a one bottle bonding in order to determine the best material for the sealing of the fissure. The depth of penetration of fuschine dye as well as that of the tested material was measured with a grid. The results, compared to the depth of the fissures, are expressed in percentage of penetration. The results were as follows: penetration of fuschine dye: 0% for the 2 composites, 100% for Helioseal F; penetration of the materials: 96.90% for Helioseal F, 70.82 for Tetric and 86.10 for Tetric Flow (significant difference, Wilcoxon test = 0.0105). In this first in vitro study, Tetric Flow shows no microleakage and is more efficient when compared to Helioseal F and Tetric in obturating deep fissures of non carious bicuspids. The second experiment of the study evaluated the microleakage and the penetration depth of Tetric Flow when it is bonded by two different methods: Group 1: total etch (phosphoric acid) and Scotch-bond 1 (3M), and Group 2: self-etching primer with Prompt (Espe). There was no significant difference (p > 0.03) between classical bonding vs self etching primer. The self-etching primer Prompt is very efficient vs phosphoric acid in obturating the fissures of non carious bicuspids with Tetric Flow. It is concluded that for prevention by sealing, using a flowable ceromer (Tetric Flow) with the self-etching (Prompt), is a really good technique. PMID- 11874010 TI - Conscious sedation in the 21st century. AB - Most (99%) patients treated at this university clinic do not need any form of sedation as rapport and behavioral management skills are more than sufficient and are safe. Those aged 1 to 5 years, who needed the use of oral sedation (Midazolam), showed 70% success. Those who needed nitrous oxide / oxygen ranged in age from 8 to 18 years and were later treated without any sort of conscious sedation after one or two sessions of nitrous oxide / oxygen sedation. PMID- 11874011 TI - Clinical evaluation of a patient with single maxillary central incisor. AB - Hypodontia in permanent dentition is the most common developmental anomaly and frequently found in the second premolar and maxillary lateral incisor In the primary dentition, however, hypodontia appears to be less frequent, with the exception of cases such as ectodermal dysplasia and cleft lip and palate. We report a child with one primary maxillary central incisor at midline. The presence of a single permanent maxillary central incisor was also confirmed by radiological examination. Other intraoral abnormalities were detected including absence of upper labial frenulum and abnormal palatal structure, but no other facial or brain anomalies. Although the condition is exceedingly rare, a thorough examination for more serious anomalies should be conducted since it is suggested to be the mildest feature of holoprosencephaly. PMID- 11874013 TI - External root resorption of the maxillary permanent incisors caused by ectopically erupting canines. AB - In this study, we analyzed the root resorption of 10 maxillary permanent incisors (two central and eight lateral incisors) in seven cases associated with ectopic eruption of adjacent canines. Two incisors were extracted because of marked root resorption. Two erupted after traction, whereas, five erupted after surgical exposure or without any treatment, although one lateral incisor submerged due to ankylosis. This suggests that self-correction of the ectopic canine occurs in some cases. PMID- 11874012 TI - Dentigerous cyst associated with an upper permanent central incisor: case report and literature review. AB - Traumas to deciduous teeth may have severe consequences. This article addresses a dentigerous cyst case report associated with an upper permanent incisor, unusual site of occurrence, which was impacted and dislodged from its natural site of eruption after trauma at the deciduous predecessor. The main aspects of etiopathology and its clinical characteristics are also discussed, with special focus on the radiographic features of the diagnosis in order to allow for an accurate surgical planning. PMID- 11874014 TI - Early detection of dens evaginatus appearing on the premolars and clinical management: histological study. AB - A review of the methods for the prevention of central cusp fracture is presented, with a case of central cusp protection of a premolar by self-curing acrylic resin by early detection. The anomalous tooth was extracted for histological observation. Closure of the pulpal horn, could be accomplished by early detection and adequate treatment for the central cusp fracture of premolars. In this case, if the gradual attrition of a central cusp were not possible, a protecting the cusp using a self-curing acrylic resin would be an effective form of treatment. PMID- 11874015 TI - Surgical treatment of mucocele in an 11 month-old baby: a case report. AB - The present report describes the occurrence of mucocele in the lower lip of an 11 month-old baby. The treatment instituted was the excision of the lesion. Microscopic examination confirmed the diagnosis of mucocele. Three months after surgery, no sign of recurrence was observed. PMID- 11874016 TI - Mobius syndrome: a case report. AB - Mobius Syndrome is characterized by showing unilateral facial nerve palsy of the sixth and seventh nerves, lack of facial expression, inability to smile and to tightly close the right eyelids. In this report, a 7-year-old-boy with Mobius syndrome is presented. He had asymmetry of facial expression, anomalies of fingers and severe tooth decay. After dental treatment, the periodic re-care visits should be done according to the eruption pattern. PMID- 11874017 TI - Orienting interns to being second-year residents. AB - BACKGROUND: Although orientation to the first year of residency is universal in family practice residencies, there is no published information on preparing first year residents to become upper-level residents, an almost equally dramatic change. METHODS: We conducted a national survey of all 484 family practice residency directors by e-mail and regular mail. RESULTS: The total response was 365 (75.4%). Slightly more than half (185) of the residencies had a formal program to prepare for the transition to second-year resident. Both faculty, and upper-level residents taught or exchanged information using multiple methods of teaching but mostly lectures and seminars. Topics included practical issues related particularly to new responsibilities, teaching and supervising skills, and the general topics of leadership and communication. An important minority objected to these orientations, expressing concern that they distracted from the ultimate goal of training competent practitioners. CONCLUSIONS: Orientation to the second year of residency is common, brief, and specific. No evaluation of the value of these programs exists. PMID- 11874018 TI - Family practice residents' awareness of medical care costs in British Columbia. AB - BACKGROUND AND OBJECTIVES: Health economics continues to be an important issue, and past studies in the United States and Europe have found that physicians and physicians in training have a limited understanding of medical care costs. No medical care cost-awareness studies have been done in Canada. In this study, the costs of 46 commonly used diagnostic tests and therapeutics were determined, and family practice residents' awareness of these costs was assessed. METHODS: Ninety seven first- and second-year residents of the University of British Columbia Family Practice Program were surveyed using the modified Dillman Total Design Method. Resident cost estimations were considered correct if within 25% or 50% of actual costs, and awareness was correlated with training location, gender, residency year, and importance ratings for ordering behavior. Degree of error was assessed by calculating median percent errors and confidence intervals for each therapeutic and diagnostic test. RESULTS: Costs were determined from the British Columbia Medical Association Guide to Fees, British Columbia Centre for Disease Control, hospital finance departments, and pharmaceutical wholesalers. A total of 82 (85%) residents completed the survey, but 11 were only partially completed. Few residents could estimate the cost of diagnostic tests or therapeutics to within 25% of the true cost, and the estimations were highly variable. Residents underestimated the cost of expensive drugs and overestimated the cost of inexpensive drugs. There was no relationship between cost awareness and training location, gender, residency year, or residents rating cost as important in ordering behaviour. CONCLUSION: Resident physicians in British Columbia, Canada have limited awareness of medical care costs. PMID- 11874019 TI - Family medicine clerkship web sites: the state of the art. AB - BACKGROUND AND OBJECTIVES: The use of on-line syllabi to acquire information about a course, clerkship, or rotation is growing. This study investigated the number of existing family medicine clerkship Web sites with syllabi and determined their completeness and usability. METHODS: In sping 1999, all schools with third-year family medicine clerkship Web sites were identified (n = 71) using Association of American Medical Colleges and American Academy of Family Physicians clerkship directories to identify required family medicine clerkships, followed by a Web search to identify any associated Web sites. A checklist with 11 content and 5 technical categories was developed and pilot tested. All sites were evaluated by at least two investigators. Two identifiable checklist characteristics were required to be present for a site to be identified as an on line syllabus and included in the study. RESULTS: Of the 71 sites initially identified, 40 were included in the study, 22 were excluded, and 9 required further review by an additional evaluator. A total of 44 sites were included in the study. The most common content features identified were instructor information, goals, overview, and requirements. Less-common features included objectives, learning resources, schedule, and grading policy. CONCLUSIONS: A minority of family medicine clerkships have on-line syllabi. Most sites have basic information, but other relevant clerkship information is absent. New and revised family medicine clerkship on-line syllabus development would benefit from using the checklist to guide the creation of instructionally sound educational resources. PMID- 11874020 TI - Physician attitudes and the use of office-based activities for tobacco control. AB - OBJECTIVES: This study explored family physicians' attitudes about smoking cessation counseling-its importance, their confidence in their ability to counsel, outcome expectations of counseling, perception of their influence on patient behavior types of counseling skills used, and the extent to which office based activities are used to support their counseling. METHODS: A cross-sectional design using qualitative and quantitative analyses was used. Data, including information from participant observation of the environment, medical chart reviews, and in-depth interviews, were collected from 89 physicians, drawn randomly from a list of family physicians in Nebraska. RESULTS: All physicians felt that counseling was important, and most were confident with their ability to provide cessation counseling. Only one third of physicians had positive expectations regarding the outcome of this counseling or of their influence on patient behavior in general. The counseling skills most likely to be used were giving advice to quit, prescriptions for pharmaceutical aids, and discussing barriers and resources. Office-based strategies to support physician counseling were seldom used. CONCLUSIONS: Physician attitudes and tobacco-control activities present a complex picture of low expectations, little office support, and limited counseling skills combined with a strong belief in the professional responsibility to counsel. Motivation to increase skills or implement supportive systems could be expected to be low. PMID- 11874021 TI - Prenatal alcohol intake in a rural, caucasian clinic. AB - BACKGROUND AND OBJECTIVES: This study examined prior use and psychosocial factors associated with alcohol and/or drug use in pregnant women from a predominantly Caucasian, rural clinic in northeastern Maine. METHODS: We conducted archival record reviews of 217 pregnant women who delivered at the Family Practice Clinic of Eastern Maine Medical Center As part of the standard initial prenatal visit during thefirst trimester, a nurse practitioner interviewed and collected data from pregnant women concerning pre-pregnancy and current-pregnancy use of alcohol, tobacco, and other drugs. Data were available for 212 subjects. RESULTS: The reported prevalence of pre-pregnancy alcohol abuse in this sample was 25%. Women in this cohort reported a significant decrease in tobacco and alcohol use following pregnancy awareness. However, pre-pregnancy alcohol intake levels and years of alcohol use were associated with alcohol intake during pregnancy. Other markers of maternal alcohol intake during pregnancy included tobacco use patterns and history of drug use. Family history of alcohol problems and drug use were associated with maternal substance use history and use by the father of the baby. Levels of maternal alcohol use during the current pregnancy were negatively associated with an alcohol problem in the father of the baby. CONCLUSIONS: Alcohol and other substance use were relatively common in our sample of rural Caucasian women in Maine. Several risk factors can be identified, and awareness of these risk factors may assist physicians in the diagnosis of substance abuse among pregnant women. PMID- 11874022 TI - A qualitative assessment of one cohort from the University of North Carolina Family Medicine Faculty Development Fellowship. AB - OBJECTIVES: This project qualitatively assessed the critical components and global impacts for one cohortfrom the part-time faculty development fellowship at the University of North Carolina at Chapel Hill. METHODS: Sixteen individual qualitative in-person interviews were conducted on one complete cohort of faculty development fellows. RESULTS: The data aggregates into four areas of influence. The first two are the critical components of the fellowship: the conditions and the curriculum. The other two are the impacts of the fellowship: the fellows' clarity of career vision and confidence. CONCLUSIONS: Graduates reported that their fellowship vitalized them to pursue their ideal position in academic family medicine. PMID- 11874023 TI - Geography and geographic information systems in family medicine research. AB - Understanding spatial relationships between determinants and outcomes of health care is important as the concept of population-based health care gains acceptance. A wide range of tools for understanding these spatial relationships is available to the family medicine researcher through the use of Geographic Information Systems (GIS). The power of GIS lies in its ability to display the spatial distribution of a health-related predictor or outcome. These maps can then be used to either generate or test hypotheses that would not have otherwise occurred to the investigator without visualizing the spatial relationships. The type of GIS application used is dependent on the type of data the researcher has and the research question. The three most common types of data are point or event data, lattice data, and geostatistical data. Point or event data can be displayed using a technique known as geocoding. Lattice data is most commonly displayed as shaded or colored areas where the shading represents rates or counts. Geostatistical data provides counts or numbers at a given location. The analytic techniques used for analyzing spatial data depend on the type of data. Maps tell powerful stories and display relationships that may not be obvious using other techniques. PMID- 11874024 TI - What authors need to know about the way editors think: report from the congress on peer review in biomedical publication. PMID- 11874025 TI - Teaching information mastery: the case of baby Jeff and the importance of Bayes' theorem. PMID- 11874026 TI - How long should a clerkship be? PMID- 11874028 TI - Support services and products available for community preceptors. PMID- 11874027 TI - Partnership created with a residency and an MPH program. PMID- 11874030 TI - Luke. PMID- 11874029 TI - Practice management: a third-year clerkship experience. AB - BACKGROUND AND OBJECTIVES: This report describes a curricular experience that introduces the basics of practice management to third-year family medicine students. The curriculum includes evidence-based medicine, managed care concepts and terminology, financial management terms and concepts, laws and legal issues, and insurance and coding. METHODS: In 1999-2000, a three-session practice management curriculum was implemented in five family medicine third-year clerkship rotations at the University of North Carolina at Chapel Hill, Wilmington site. The learning activities included readings, a "quiz show" game, and a patient practice-centered project. RESULTS: Students reported greater knowledge and confidence in the selected topics. The practice management curriculum was positively evaluated overall. CONCLUSIONS: This interactive learning module for the family medicine students begins the process of incorporating practice management skills and knowledge. PMID- 11874031 TI - The doctor: a seminal video for cinemeducation. PMID- 11874032 TI - Applications of narrative theory and therapy to the practice of family medicine. AB - This article presents narrative theory and therapy as an approach with significant potential for providing family physicians with additional tools to assist them in dealing with difficult clinician-patient encounters. We first define narrative therapy, then briefly describe its theoretical assumptions in relation to psychosocial concepts already familiar to family physicians. Important aspects of narrative therapy are examined, including the unique role of questioning in the narrative process; understanding and helping patients change their problem-saturated stories; renaming and externalizing the patient problem; and the use of rituals, documents, and audience in recognizing and reinforcing patient change. The article concludes with thoughts about how narrative approaches can contribute to more-healing doctor-patient relationships. PMID- 11874033 TI - Heavy metals in emergent trees and pioneers from tropical forest with special reference to forest fires and local pollution sources in Sarawak, Malaysia. AB - Leaf samples of tropical trees, i.e. Dryobalanops lanceolata (Kapur paji), Dipterocarpaceae and Macaranga spp. (Mahang), Euphorbiaceae were analyzed for 21 chemical elements. The pioneer Macaranga spp. exhibited higher concentrations for the majority of elements compared to the emergent species of Dryobalanops lanceolata, which was attributed to the higher physiological activity of the fast growing pioneer species compared to emergent trees. Lead showed rather high concentrations in several samples from the Bakam re-forestation site. This is suggested to be caused by emissions through brick manufacturing and related activities in the vicinity. A comparison of Dryobalanops lanceolata samples collected in 1993, 1995 and 1997 in the Lambir Hills National Park revealed that certain heavy metals, i.e. Co, Cu, Mn, Ni, Pb and Ti showed higher values in 1997 compared to the previous years, which could indicate an atmospheric input from the haze caused by the extensive forest fires raging in Borneo and other parts of Southeast Asia. PMID- 11874034 TI - Dietary intake of cadmium by children and adults from Germany using duplicate portion sampling. AB - The dietary intake of cadmium was studied among children and adults from Germany. The study area comprised the highly industrialized Ruhr district as well as the North Sea island Amrum. A total of 229 duplicate samples were collected from 49 individuals between December 1994 and May 1995. The sampling period for each participant was either 3 or 7 days. Cadmium concentrations in duplicate samples were measured by atomic absorption spectrometry. The daily cadmium intakes (geometric mean) for 4 different groups of individuals distributed as follows: for small children (mean age: 1.8 years) from the Ruhr district 0.17 microg/(kg(bw) x day) [range: 0.02-1.62 microg/(kg(bw) x day)], for children (mean age: 3.8 years) from the Ruhr district 0.49 microg/(kg(bw) x day) [range: 0.11-2.06 microg/(kg(bw) x day)], for children (mean age: 3.9 years) living on the North Sea island Amrum 0.35 microg/(kg(bw)-day) [range: 0.09-1.13 microg/ (kg(bw) x day)] and for adults (mean age: 40.9 years) from the Ruhr district 0.37 microg/(kg(bw) x day) [range: 0.05-1.32 microg/(kg(bw) x day)]. Compared to the provisional tolerable weekly intake (PTWI) of 7 microg/(kg(bw) x week) proposed by the WHO the dietary intake of cadmium was rather high. The geometric mean and maximum intake values for the different groups ranged between 24.3-55.7% and 62.7 120.7 respectively of the PTWI. We therefore conclude, that the cadmium exposure of the population needs to be reduced in order to minimize the risk of adverse health effects related to this metal. PMID- 11874035 TI - Spatial and temporal variation of polychlorinated biphenyl (PCB) in precipitation in southern Sweden. AB - Spatial and temporal variations in polychlorinated biphenyl (PCB) concentrations were studied in precipitation at 11 sampling-sites over a period of 1 year and compared with PCB concentrations in the air. The study was carried out in a region of southern Sweden approximately 11 000 km2 in area. The PCB concentration in precipitation ranged between 1.18 and 81.4 ng l(-1). Two of the sampling sites showed concentrations that were approximately 30 times higher than at the rest of the sites. The variation in concentration between sites may be explained by location as well as by differences in weather conditions and particle content in the air. Wind direction played an important role for PCB concentration in precipitation in coastal areas, while at the inland sites this variable seemed to have a minor influence. We found no seasonal trends in PCB concentration in precipitation or any dependence on air temperature. To examine the intensity of precipitation scavenging, the total washout ratios (Wt) were calculated. There was a large variation in Wt over time, and the highest ratios were observed at the two sites where PCB concentration in the air was high. Furthermore, high concentrations of PCB in precipitation correlated with a high-chlorinated pattern, as shown by principal component analysis. For most of the sites there was a significantly negative relationship between PCB concentration and rain volume. PMID- 11874036 TI - Role of microbes in controlling the speciation of arsenic and production of arsines in contaminated soils. AB - The influence of microbes on the speciation of arsenic and production of arsines in contaminated soils was investigated under laboratory conditions. Microbes were able to carry out reactions that resulted in changes in the speciation of arsenic in soil. The transformation of soil dominating species, arsenate [As(V)], under both aerobic and anaerobic conditions to arsenite [As(III)], monomethylarsonic acid [MMAA], dimethylarsinic acid [DMAA] and to volatile trimethylarsine [TMA] was, however, less than 0.5%, of which the production of TMA represented 0.02 0.3%. The volatilization process was also verified in the field, in the soil of a dumping area. The 'life-time' of arsines in air was, however, short and they were rapidly converted back to water soluble species, As(V) and trimethyl arsine oxide (TMAO). PMID- 11874037 TI - Dispersion of PCB in the environment following an atmospheric release caused by a fire. AB - During a fire at a power plant located in the coastal plain of Israel, PCBs were released to the atmosphere from a ruptured transformer. Since PCBs are probably carcinogenic to humans, this study was performed in order to assess the environmental contamination by PCBs via the atmospheric pathway and the need for remediation measures. The release conditions and the meteorological conditions which prevailed during the fire were analyzed. This provided the input to a Gaussian dispersion model used to estimate the downwind-contaminated sector as well as the location of the maximal concentration within this sector. A sampling plan was then devised and vegetation collected within this sector was analyzed for PCBs. A methodology was developed to convert PCB concentrations in vegetation to concentrations in the atmosphere. It allowed a reconstitution of the PCB source term to the atmosphere from the vegetation measurements. The PCB concentrations were found to be lower than the USEPA decontamination standards. Remediation measures were not needed beyond the plant fence. PMID- 11874038 TI - Trace element analysis of Cretan wines and wine products. AB - The object of this research is to investigate the ways and the degree of contamination of Cretan grapes from the area of Chania and their alcoholic products, with the elements aluminium, arsenic, cadmium, copper, chromium, iron, lead, manganese, nickel and zinc. Fifteen samples of grapes were collected and used for the production of experimental wines from rinsed and unrinsed grapes. A microwave furnace was used for the digestion and dissolution of the experimental wines, the precipitates that originated in these wines, as well as the wines of the corresponding producers. The analyses of all mentioned samples as well as 34 local alcoholic distillates were performed using total reflection X-ray fluorescence and graphite furnace atomic absorption spectrometry. The concentrations for all the elements that were determined were almost in all cases, well below the maximum permissible levels by the Greek and the European Union legislation. PMID- 11874039 TI - Chemical quality of bottled waters from three cities in eastern Alabama. AB - Twenty-five brands of bottled waters consisting of both purified and spring types collected randomly from three Alabama cities, USA were assessed for their suitability for human consumption. Water quality constituents analyzed include pH, conductivity, alkalinity, chloride, nitrate + nitrite, sulfate, phosphate, total carbon (TC), inorganic carbon (IC), total organic carbon (TOC), and 27 elements on the inductively coupled plasma-optical emission spectrometer (ICP OES). The results obtained were compared with US EPA drinking water standards and the European union (EU) drinking water directive. Ni was non-detectable in all the samples and Cu, Pb, Sb, Zn, Mn, Al, Cr, Mg, P, Ca, sulfate, chloride and nitrates + nitrites were all below their respective USEPA drinking water standards or EU maximum admissible concentrations (MAC). The conductivity, pH, As, Cd, Hg, Zn, Se and Tl values in some samples exceeded the EU and USEPA standards for drinking water. No sample had pH > 8.5, but seven bottled water brands analyzed were acidic (pH < 6.5). Most of the sampled brands had TOC concentrations exceeding 3 mg/l. The concentrations of most water quality constituents analyzed, in most cases, were higher in the spring water brands compared to the purified or distilled brands of bottled water. A one-way parametric analysis of variance (ANOVA) conducted on pH, conductivity, IC, TOC, Ca, Na, K, Mg, Se, sulfate, chloride and nitrate + nitrite values for 10 brands of bottled water to ascertain the homogeneity of variances within and between the brands, suggested significant differences in variances across the brands at a 95% confidence level except for selenium, sodium and calcium. PMID- 11874040 TI - Daily intake of TBT, Cu, Zn, Cd and As for fishermen in Taiwan. AB - The consumption of contaminated seafood has been reported as an important route of human exposure to metals in Taiwan. We consider the concentrations of TBT, Cu, Zn, Cd, As, and the consumption of oysters of Taiwanese to be the important information related to public health in Taiwan. Therefore, the aim of this study was to evaluate the public health risks associated with TBT, Cu, Zn, Cd and As from shellfish for the general population and fishermen of Taiwan. In general, TBT concentrations in various oysters ranging from 0.32 to 1.51 microg/g dry wt. varied with sampling locations. The highest TBT, Cu, and Zn geometric mean (GM) concentrations in oysters of 1.51, 1180 and 1567 microg/g dry wt. were obtained from the Hsiangshan coastal area. The values of oyster consumption for fishermen were 94.1 and 250 g/day for typically and maximally exposed individuals, respectively. In particular, the highest intake (250 g/day) from fishermen was almost two times greater than that of the general population (139 g/day). The THQ (target hazard quotient) values of Hsiangshan's fishermen are 3.87 and 20.50 for TBT and Cu for maximally exposed individuals are higher than other oyster culture areas. It is interesting that those consuming oysters from Hsiangshan, Lukang, Taishi caused abnormally high THQs of TBT and other metals (100% over 1.0), and TBT was attributed to only 3-21% of the total THQs in different fishermen of Taiwan. Our results suggest that current environmental levels of TBT and other metals are associated with a significant potential threat to human health for fishermen resident in coastal areas of Taiwan. PMID- 11874041 TI - Cadmium and zinc interactions and their transfer in soil-crop system under actual field conditions. AB - The transfer of Cd and Zn from calcareous soils nearby a non-ferrous mining and smelting bases to the spring wheat (Triticum aestivum L.) and corn (Zea mays L.) tissues and the interactions between the two metals concerned were investigated under actual field conditions. Samples of soils and entire crops were randomly collected during harvest time in 1998 in the Baiyin region. The soil metal contents showed that the furrows had been polluted (mean values: 3.16 mg kg(-1) for Cd; 146.78 mg kg(-1) for Zn) and the significant spatial variation of the soil contamination existed here (ranges, Cd: 0.14-19.3 mg kg(-1); Zn: 43.5-565.0 mg kg(-1)). The translocation ratios of the two metals from soil to crop parts in the region studied were relatively lower and the order of the element transfer in different plant tissues was root > stem > grain. The transfer ratio of element Cd was lower than that of element Zn. Cd and Zn uptake by the crop structures could be best described by four models (P < 0.01): linear; exponential; quadratic; and cubic. Apart from a linear relationship between the element Cd in the corn grains and soils, models were generally non-lincar. An analysis of Cd-Zn interaction mechanism led to the conclusion that the effects of the two metals were synergistic to each other under field conditions, in which increasing Cd and Zn contents in soils could increase the accumulations of Zn or Cd in the two crops. PMID- 11874042 TI - Distribution of some trace metals in Lochnagar, a Scottish mountain lake ecosystem and its catchment. AB - Anthropogenic trace metals enter the entire ecosystem of Lochnagar solely through atmospheric deposition. Trace metals, including Hg, have been monitored in atmospheric deposition and lake water, and measured in catchment vegetation, aquatic plants and zooplankton, revealing contamination levels in the ecosystem. Furthermore, 17 sediment cores were taken from different areas of the lake. Hg, Pb, Cd, Zn and Cu were analysed in all the cores, which show that the sediments have been heavily contaminated by these trace metals since the 1860s. The distribution of trace metals in the lake sediments was found to be heterogeneous, with concentrations in the surface sediments varying significantly: 110-250 ng/g, 100-360 microg/g, 39-180 microg/g, 0.3-1.9 microg/g and 8-25 microg/g for Hg, Pb, Zn, Cd and Cu, respectively. Trends in the concentration profiles for different trace metals in the same core are different, as are the trends of the profiles for the same metal in different cores. Hence, a single sediment core cannot represent the pollution history of the whole lake. As the soils and sediments contain a high proportion of plant debris and the debris has a high affinity for Hg, resulting in Hg enrichment. Hg was measured in plant debris (> 63 microm) separated from catchment soils and lake sediments. Plant debris may play an important role in storing and transferring Hg in this ecosystem. PMID- 11874043 TI - Measurements of particulate matter within the framework of the European Monitoring and Evaluation Programme (EMEP) I. First results. AB - Particulate matter (PM) monitoring presents a new challenge to the transboundary air pollution strategies in Europe. Evidence for the role of long-range transport of particulate matter and its significant association with a wide range of adverse health effects has urged for the inclusion of particulate matter within the European Monitoring and Evaluation Programme (EMEP) framework. Here we review available data on PM physico-chemical characteristics within the EMEP framework. In addition we identify future research needs for the characterisation of the background PM in Europe that include detailed harmonised measurements of mass, size and chemical composition (mass closure) of the ambient aerosol. PMID- 11874044 TI - Selenium levels in biological matrices in adult population of Mumbai, India. AB - Selenium (Se) levels in whole blood, serum, urine, muscle and saliva of Mumbai adults have been estimated by differential pulse cathodic stripping voltammetry (DPCSV); the detection limit of Se is 0.05 ng ml(-1). The reliability of estimation is further assessed through the analysis of Standard Reference Materials. The Se levels in whole blood (n = 35) and blood serum (n = 201) of the Mumbai adult population is 99.6 and 100 ng ml(-1); approximately 34.8% of the population have serum Se levels between 80 and 100 ng ml(-1). The blood serum levels of Se for Mumbai adults are comparable to those of whole blood. The mean concentration of Se in urine on a 24-h basis is 5.2 ng ml(-1). Muscle and saliva of the Mumbai adult population contain 195.4 ng g(-1) and 2 ng ml(-1) of Se, respectively. A good correlation between serum Se and dietary Se is observed with a correlation coefficient of 0.89. PMID- 11874045 TI - The stability and persistence of diflubenzuron in marine sediments studied under laboratory conditions and the dispersion to the sediment under a fish farm following medication. AB - A high performance liquid chromatographic (HPLC) method was developed to determine the concentration of diflubenzuron, a delousing agent used in fish farming, in marine mud and shell sand. The recovery of diflubenzuron from mud was 100.8+/-1.1% and 105.5+/-4.3% for shell sand. The limit of quantitation was found to be 0.1 microg g(-1). The stability of diflubenzuron was studied under laboratory conditions in marine sediments at different temperatures (4 and 14 degrees C). No degradation of diflubenzuron occurred in the organic rich mud sediment or in the shell sand sediment during the experimental period of 204 days. Increasing the temperature from 4 to 14 degrees C had no effect on the stability. Furthermore, diflubenzuron showed to be persistent in both mud and shell sand sediment since no detectable diffusion from the sediment to the water phase occurred during the experimental period of 204 days. Increasing the water current in the tanks had no effect on the persistence. Under field conditions, the concentrations of diflubenzuron found in the organic material from sediment traps placed 2 m from the bottom under the cage in a fish farm during medication were high and ranged from 71 to 259 microg g(-1). The concentrations of diflubenzuron in the sediment under the fish farm were, however, low, with a maximum concentration of 5.4 microg g(-1). The dispersion of diflubenzuron to the sediment was limited to less than 20 m from the edge of the cage in every direction. Fifteen months following the medication, only traces (< 0.1 microg g( 1)) of diflubenzuron were detected in the sediment under the fish farm. Possible explanations for this decrease are resuspension and redistribution of sediment and/or oxic degradation of the drug. PMID- 11874046 TI - Solid waste: terminological and long-term environmental risk assessment problems exemplified in a power plant fly ash study. AB - Legal definitions exert a significant impact on the waste management strategy. Waste that is technically suitable for recovery does not automatically become a raw material if there is no market for it, or its use is not commercially effective and, hence, they should be disposed of. The majority of disposed wastes, including recyclable waste, are not environmentally safe. Waste as a freshly generated anthropogenic material is not geochemically stable. Przezchlebie fly ash surface pond (Upper Silesia, Poland) in the post-closure stage was subject to field validation of the results of laboratory leaching/extraction tests and long-term column experiments on fly ash (FA) leaching behaviour under controlled conditions for environmental risk assessment. The study showed: (i) the possibility of a discontinuous non-linear time delayed increase of pollution potential of disused 'non-hazardous' large-volume waste in the dumping sites to the hazardous level; (ii) inconsistency of the laboratory leaching tests and the actual leaching behaviour of trace metals, particularly when equilibria conditions are dictated by kinetically determined reactions where the test results reflected entirely wash-out (I) and dissolution (II) phases, but did not comprise delayed release (III) phase; and (iii) necessity of life-cycle screening/monitoring of 'non-hazardous' dumping sites for contaminant release as a function of the primary (pH-Eh, ionic strength, ionic composition of solute) and secondary controlling factors (L/S-liquid to solid ratio, water flow conditions) along the vertical profile of an anthropogenic or natural vadose zone. These data are to be used to develop long-term predictive hydrogeochemical models and their field validation, and for providing an early warning and remedial actions with respect to the particular site. The formation of pH (and Eh) as a function of time-dependent (kinetically defined) processes appeared to be a key issue for a correct prediction of the leaching behaviour of waste. The presented case study on FA shows that waste, even those considered non-hazardous and fit for use in a commercially proven applications, should not be treated the same way as a natural raw material. To facilitate waste utilisation and disposal in environmentally safe way and to prioritise its use, reliable environmental risk assessment prediction models and testing procedures, as well as special enforcement strategy and regulations, should be developed with respect to waste and not 'materials.' PMID- 11874047 TI - Influence of nonylphenol on the microbial community of lake sediments in microcosms. AB - In this study the impact of nonylphenol, an estrogenic degradation product of alkylphenol polyethoxylates, on the microbial community structure in contaminated sediments of aquatic microcosms using in situ hybridization with fluorescently labeled oligonucleotides probes was investigated. A positive correlation between nonylphenol concentration and cell numbers of bacteria and microfungi as well as an increase in the numbers of active bacteria was found. However, the ratio between total microorganisms and active bacteria remained unchanged. A large fraction of the cells could be identified using group specific oligonucleotide probes. A slight change in the composition of the microbial community structure was observed, with Gram-positive bacteria with high DNA G + C-content becoming more abundant at higher concentrations of nonylphenol. PMID- 11874048 TI - Exchange of organic solvents between the atmosphere and grass--the use of open top chambers. AB - Volatile organic compounds (VOC) are of increasing environmental significance as a result of continually increasing volumes of traffic on European roads. An open top chamber fumigation system has been devised to investigate how these contaminants transfer between the atmosphere and the ground, and how they partition between and within air-plant-soil systems. Variation in chamber temperature, solar radiation in the chamber and chamber flow rate were identified as factors that affected final air concentrations. These were assessed and quantified for all individual chambers used--effectively characterising each chamber. The real-life VOC concentrations generated were stable and readily reproducible. Grass exposed to benzene, toluene, 1,1,1-trichloroethane and tetrachloroethene, respectively, equilibrated in response to a change in air concentration within hours. The rate of equilibration in exposed grass in all cases was independent of air temperature. 1,1,1-Trichloroethane and tetrachloroethene appear to be biologically inert demonstrating a simple physico chemical approach to equilibrium, however, benzene and toluene do not appear independent of plant metabolic activity. Aqueous solubility can account for all of the toluene and benzene in the fumigated plant material. PMID- 11874049 TI - Influence of irrigation with lagooned urban wastewater on chemical and microbiological soil parameters in a citrus orchard under Mediterranean condition. AB - The reduced availability of water resources in semi-arid Mediterranean regions requires an efficient use of supply sources. Urban wastewater, after treatment to minimise health hazards, may constitute an important resource for irrigation in areas characterised by intensive agriculture. These considerations have motivated an investigation (during the irrigation season 1996) of the dynamics of microbial biomass in the soil of a citrus orchard in eastern Sicily, which has been irrigated for 15 years with lagooned urban wastewater, to evaluate the effects of this practice on soil fertility. The analyses of parameters regarding soil microbial biomass (microbial carbon and microbial nitrogen, soluble carbon and nitrogen, cumulative respiration, respiratory quotient and enzymatic activity in the soil) have confirmed that the evolution of soil microflora is directly conditioned by the type of water used for irrigation and climatic conditions. Just before the beginning of the irrigation season (May), microbial biomass carbon (MBC), soluble C, cumulative respiration and enzymatic activity were significantly higher in the soil irrigated with wastewater with respect to the same soil irrigated with 'clear' water; the qCO2 was significantly lower. In June, after 1 month of irrigation, both soils showed an increase of all microbial parameters except for enzymatic activity and qCO2 which showed a significant reduction. In September, at the end of the irrigation period, there was a decrease of almost all investigated parameters in both plots, which was more evident in the soil irrigated with 'clear' water. The microbial biomass of the soil irrigated with wastewater, during the irrigation period, did not undergo any negative effects, having an evolution analogous to the plot irrigated with 'clear' water. The use of lagooned wastewater after three lustrums has shown, particularly in the dry season, an increase in quantity of easily available nutrients, with an improvement of the metabolic efficiency of soil microflora coupled with a more marked activity of total hydrolase and phosphatases. The variations of the parameters related to the soil microflora were strongly influenced by the seasonal climatic trend. PMID- 11874050 TI - The status and trends of trace element and organic contaminants in oysters, Crassostrea virginica, in the waters of the Carolinas, USA. AB - Concentrations of eight trace elements (As, Cd, Cu, Hg, Ni, Pb, Se and Zn), lindane and six groups of organic contaminants (total-chlordane, total-PCB, total DDT, Dieldrin and Aldrin, total-butyltins, total-PAHs) at the 11 NOAA mussel watch project (MWP) sites located in North and South Carolina have been compared with the national US MWP data. Three sites from North and South Carolina had concentrations of PAHs in the upper 15th percentile on a national scale. One site had high concentrations of butyltins, and two sites had high Se concentrations. All sites from Beaufort, North Carolina, south had high As concentrations. Decreasing temporal trends were found for As, Cd, total-chlordane, DDT, PCB, and PAHs at some sites. PMID- 11874051 TI - Effect of smoking and alcohol consumption on the serum selenium level of Lower Silesian population. AB - Serum selenium (Se) levels and glutathione peroxidase (GSH-Px) activity in erythrocytes have been determined for a healthy Lower Silesian population. The results have been analysed in relation to smoking and alcohol consumption habits. The mean concentration of serum Se for the group is within the lower range and appears to be significantly lower in the group of smokers that was studied (smoking over 20 cigarettes/day, mean concentration value = 50.8 microg/l), compared to non-smokers (mean value = 78.8 microg/l). For men that smoke, the concentration is lower than that found for smoking women, however, the difference is without statistical significance. Regarding GSH-Px activity, no difference between the smoking and non-smoking groups has been found (mean values = 19.84 and 20.92 U/gHb, respectively). From the group of people examined, we have selected those who drink an equivalent of 50 g or more of pure ethanol per week. For this group the serum Se level has been found to be lower compared to the group that does not consume that much alcohol. However, the difference is without statistical significance. We have found no correlation between age and the parameters studied (serum Se concentration and GSH-Px activity). PMID- 11874052 TI - Child dental expenditures: 1996. AB - PURPOSE: Because little has been reported about child dental expenditures, federal data were used to estimate dental care expenditures for U.S. children by age, sex, ethnic/ racial background, family income, parental education and parental employment. METHODS: Parentally reported data on dental expenditures and sources of expenditures were extracted from the most recent available federal healthcare expenditures studies, the 1996 federal Medical Expenditure Panel Survey (MEPS). Using the survey's large sample and complex design, these data represent the entire U.S. child population. RESULTS: Nearly 12 billion dollars were expended for children's dental care averaging $375 per child who obtained care. Overall sources of payment were 47% out of pocket, 45% insurance and 8% "other" including primarily Medicaid. Disproportionately litde spending was made on behalf of low-income and minority children despite their higher disease experience. The proportion of spending that was paid out of pocket was high for all groups of children including those eligible for Medicaid even though Medicaid prohibits cost sharing. CONCLUSIONS: Dental care for children accounts for approximately one-quarter of U.S. dental spending and is a major component of child health care costs. Income and racial disparities in expenditures favor higher income children despite Medicaid coverage for lower income children. High levels of reported out-of-pocket costs for Medicaid eligible children suggest that Medicaid fails to meet families' needs in obtaining care. Meeting the oral health needs of poor children will require considerably greater expenditures, particularly through improved Medicaid financing and administration. PMID- 11874053 TI - Effects of changing U.S. parenting styles on dental practice: perceptions of diplomates of the American Board of Pediatric Dentistry presented to the College of Diplomates of the American Board of Pediatric Dentistry 16th Annual Session, Atlanta, Ga, Saturday, May 26, 2001. AB - PURPOSE: This study surveyed board-certified pediatric dentists on their opinions about changes in U.S. parenting styles and the effects on the practice of pediatric dentistry. METHODS: A questionnaire was developed, piloted, and mailed to 1,129 members of the College of Diplomates of the American Board of Pediatric Dentistry during the summer of 2000. RESULTS: A total of 577 respondents (51%) returned questionnaires, equally distributed across AAPD districts, with male:female ratio of 4:1 and 90% married. A majority perceived parenting styles had changed during their practice lifetime (88% "absolutely or probably changed"), with older practitioners significantly more likely to say so. Ninety two percent felt changes were "probably or definitely bad" and 85% felt that these changes had resulted in "somewhat or much worse" patient behavior. Practitioners report performing less assertive behavior management techniques than in the past due to these changes. CONCLUSIONS: Diplomates report that parenting changes have occurred and they believe these are negative (bad) and have adversely influenced behavior and caused changes in pediatric dentists' behavior management. PMID- 11874054 TI - Microleakage of a new improved glass ionomer restorative material in primary and permanent teeth. AB - PURPOSE: The objective of this study was to assess the microleakage of the new conventional glass ionomer, Fuji IXgp in comparison to another conventional glass ionomer (Fuji II), a resin modified glass ionomer (Vitremer) and a composite resin (TPH) in primary and permanent teeth. METHODS: Twenty-five extracted human premolars and 13 primary molars were used. Preparations were made on the center of the buccal and lingual aspects of the premolars (Group A) and the mesio buccal and disto buccal surface as well as the mesio lingual and disto lingual surface of the primary molars (Group B). Each group was randomly divided into five subgroups of 10 specimens each and restored with a different material following the manufacturer's recommendations. Restorations were subjected to thermocycling followed by microleakage evaluation using 50% silver nitrate and computerized image analysis. RESULTS: Two factor analysis of variance revealed a significant main effect of material (P<0.001), a trend toward a main effect of tooth type (P=0.082) and a significant interaction term P=0.016. Materials were a source of difference, so a one-way ANOVA test was used for both primary and permanent teeth together and for each individual group of teeth. Differences were further examined with a multi-variate analysis using the Scheffe' test for both groups of teeth and each individual group of teeth. Each group of teeth restored with the same material was then analyzed with an Independent Samples t-test which showed that conventional glass ionomer (Fuji II) had more leakage than all other groups (P<0.01). In addition TPH showed more microleakage in primary teeth (P<0.02) and Fuji II showed more microleakage in permanent teeth (P<0.02). CONCLUSIONS: Fuji IXgp behaved similarly to the composite (TPH) and to the resin modified glass ionomer (Vitremer). This is a promising result for this material that is targeted for application in conjunction with the Atraumatic Restorative Technique and minimal intervention treatments. PMID- 11874055 TI - Partial pulpotomy for immature permanent teeth, its present and future. AB - Traumatic or carious exposure of a vital pulp in an immature permanent tooth presents a significant clinical challenge to maintain proper vitality. Currently available procedures include direct pulp capping, complete pulpotomy and partial pulpotomy. This paper reviews the application of partial pulpotomy in immature permanent teeth and provides prognostic and technique guidance. PMID- 11874056 TI - Management of caries in the child three years of age and younger: a survey of post-doctoral pediatric dentistry program directors. AB - PURPOSE: The purpose of this study was to report the results of a survey of pediatric dentistry post-doctoral program directors regarding education of post doctoral students about management of caries in children 3 years-of-age or younger. METHODS: Fifty-two pediatric dentistry advanced education program directors were sent questionnaires inquiring about payer sources in their programs, distribution of caries in children 3 years of age or younger within the payer sources, the methods they teach and use to treat the caries, and the effectiveness of treatment and outcome data about the success of their treatment. RESULTS: Twenty-nine programs responded (56%). On average, two-thirds (66%) of the patients in post-doctoral pediatric dentistry programs are Medicaid patients. Program directors are fairly uniform in how they define methods of caries management, and they rate definitive therapy as the most effective method to manage all types of caries. Literature/textbooks were most frequently cited as the major source of scientific evidence to support treatment decisions. Fewer than 20% of program directors have outcome data on the effectiveness of their methods of treatment. CONCLUSIONS: Medicaid is the major payer source for patients in post-doctoral programs and definitive therapy is considered by program directors to be the most effective approach to managing caries in this patient population. Program directors rely on the literature and textbooks and few have outcome data. PMID- 11874057 TI - The diagnostic value of lateral extraoral radiography for intruded maxillary primary incisors. AB - PURPOSE: The purpose of the study was to assess the contribution of a lateral extraoral radiograph for diagnosing the relation between the root of intruded maxillary primary incisors and their permanent successors. METHODS: Three pediatric dentists examined intruded primary teeth 0-7 days after injury. The relations between the primary and permanent teeth were assessed in three separate steps: 1) evaluation of clinical signs only; 2) evaluation of a periapical radiograph (no lateral radiograph); 3) evaluation of a lateral extraoral radiograph. The clinical and radiographic signs used to assess the relations were recorded. The lateral extraoral radiograph was regarded as contributory to diagnosis if the assessment after the third step differed significantly from that made after the first and second steps. Ninety-three evaluations of 53 intruded teeth in 37 children were available for analysis. RESULTS: The lateral extraoral radiograph was found valuable for assessment of the primary incisor's root alignment in only 5% (5/93) of the evaluations in which neither the clinical examination nor the periapical radiograph were contributory. Four of these five cases were in children less than 20 months old. In all other cases, the lateral radiograph was not contributory for two main reasons: It could not be evaluated due to overlap of multiple intruded teeth and/or when the teeth intruded were lateral incisors, and when the clinical and periapical radiographs were sufficient for diagnosis. CONCLUSIONS: Lateral extraoral radiographs should not be used routinely in cases of intrusion of primary incisors. The operator should base his or her diagnosis on clinical findings and examination of a periapical radiograph. The lateral extraoral radiograph should be taken only when its expected contribution to diagnosis can be confirmed, as in cases of children younger than 20 months. PMID- 11874059 TI - Gingival disease associated with a decorative crown. AB - Decorative crowns for the teeth have gained a resurgence of popularity among adolescents. Similar to other forms of body art found in the mouth, these trendy crowns may be associated with a variety of oral complications. This case report describes a localized form of necrotizing ulcerative gingivitis, exacerbated by a cosmetic gold crown in a teenage girl. Healthier artistic options are discussed to replace this flashy dental fad. PMID- 11874058 TI - Dental findings associated with the malformations of CHARGE. AB - The acronym CHARGE refers to a non-random clustering of congenital malformations whose cause remains unknown. To date, the dental features of CHARGE association are not well known. A brief review of the literature and a case in a 10-year old boy with the CHARGE association are presented. The patient had multiple dental anomalies including congenital absence of teeth, ectopic eruption, submergence of primary molars and an odontogenic fibroma associated with an impacted permanent molar. PMID- 11874060 TI - What a practice administrator can do for you. AB - Most dentists have at some time during their practice years yearned for administrative help. This article reasons through the why, what, who and how of employing a Practice Administrator so that the dentist is free to concentrate on clinical rather than business aspects of the practice. PMID- 11874062 TI - Dental care of the pediatric patient with splenic dysfunction. AB - The spleen is strategically situated in the vascular tree to filtrate the circulating blood, therefore it is of paramount importance in the defense against pathogenic organisms. Sepsis in individuals who have splenic dysfunction or have had a splenectomy is a life-long risk, thus patients, caretakers and health professionals should prevent infections and recognize them promptly so that treatment can be instituted immediately. This manuscript reviews the consequences of splenic dysfunction and provides recommendations for dental care of the pediatric patient. PMID- 11874061 TI - Use of self-etching adhesive system and compomer for splinting traumatized incisors. AB - Traumatized teeth that are displaced from their normal alveolar positions need to be repositioned and stabilized so that healing can occur. Orthodontic ligature wire bonded into place with resin-based composite works well for splinting teeth. Using a self-etching adhesive bonding system and compomer material facilitates both the resin bonding and its removal. This clinical report describes a simplified resin/wire splinting procedure. PMID- 11874063 TI - Stretching the safety net too far waiting times for dental treatment. AB - PURPOSE: The objective of this study was to survey pediatric dentistry program directors on perceived resource needs and changes over the last 5 years in the characteristics of their patient population and on waiting times for dental treatment with sedation or general anesthesia (GA) for children with complex dental and medical histories in hospital- and dental school-based training programs. METHODS: A 47-question survey was sent electronically to all pediatric dentistry program directors in the United States using the University of Washington's Catalyst Tools program. Two reminder messages were sent. After 3 months, the data was downloaded and descriptive statistics were performed using the SPSS for Windows version 8.0. RESULTS: Twenty-eight of 54 program directors responded with 26 usable survey responses (48%). Thirty-one percent reported outpatient clinics located in a dental school, 31% reported that their clinics were in a hospital, and 38% had clinics in both settings. Program directors perceive that the number of new, recall and emergency patients and the number of pre-school aged children and children with special health care needs had increased in their programs in the last 5 years. Payment by Medicaid was the most common insurance for children cared for in these settings. The mean waiting time for scheduling treatment with GA for a child in pain is 28 days; without pain 71 days. The mean waiting time for scheduling treatment with sedation is 36 days. The majority of program directors reported they had an adequate number of faculty and residents (61% and 66%, respectively) even though 52% of the directors were presently actively recruiting faculty. CONCLUSIONS: 1. Dental school and hospital based training programs are an important source for an increasing number of children with complex dental needs; 2. The majority of patients treated in the programs are Medicaid beneficiaries; 3. Average waiting times for complex dental care for children in pain is 28 days with GA; without pain and need for GA 71 days; 4. There was an average 36-day wait for treatment with sedation. PMID- 11874064 TI - Frequency of reported dental visits and professional fluoride applications in a cohort of children followed from birth to age 3 years. AB - PURPOSE: The purpose of this study is to report the prevalence of having a dental visit and/or a topical fluoride treatment at the dental visit from birth to age 36 months from an observational, longitudinal study of a cohort of children followed since birth. METHODS: Multiple questionnaires were sent to the families of children enrolled in a study of fluoride ingestion to ask parents if their child had had a dental (or dental hygiene) appointment and/or a fluoride treatment during the time interval since the previous mailed survey. Data were analyzed to determine the percentages of children who had at least one dental visit or at least one fluoride treatment by one, two or three years of age. RESULTS: Three hundred and forty parents completed all eleven questionnaires during the 36-month study period. Of these, 2% reported having taken their child for a dental visit by one year of age, 11% by two years of age and 31% by three years of age. Of those with a visit by three years of age, 19% received at least one fluoride treatment. CONCLUSIONS: Despite recommendations for early dental visits, only 31% of the children in this cohort had been seen by a dentist by the time they reached their third birthday and 19% of these had received a professional fluoride treatment. PMID- 11874065 TI - Failure rates of restorative procedures following dental rehabilitation under general anesthesia. AB - PURPOSE: The failure rates of restorative procedures for children undergoing dental rehabilitation under general anesthesia, performed by pediatric dental residents in advanced educational programs, were evaluated in order to determine treatment outcomes and best practices. METHODS: Retrospective review of 504 dental records of children receiving comprehensive dental treatment under general anesthesia at children's hospitals in Boston between 1990-1992 and in Washington, DC, between 1994-1998, were undertaken. Data regarding restoration outcomes were evaluated using chi square tests with correction for continuity. Only records of patients who returned for follow-up at least six months after their rehabilitations were evaluated. T-tests were performed on parametric data. RESULTS: Two-hundred and forty-one (48%) of the records were evaluated. Stainless steel crowns (SSCs) had significantly lower failure rates than amalgams (P<0.001, chi2=63). The highest failure rates were seen in composites (P<0.001, chi2=112) and composite strip crowns (P<0.001, chi2=121). CONCLUSIONS: SSCs are the most reliable restorations while composite restorations are the least durable. Failure of restorations appears to be related to follow-up length. PMID- 11874066 TI - Clinical section--case reviews: an analysis. PMID- 11874067 TI - Vulval intraepithelial neoplasia: current perspectives. AB - The heterogeneous clinical features of vulval intraepithelial neoplasia (VIN), its uncertain and variable natural history, difficulties in the management of certain cases and the frequency of recurrences provide a continuing challenge for gynaecologists. Patients with VIN present to a diverse range of physicians, all of whom provide differing perspectives on the multifarious issues relating to the condition. The importance of VIN relates principally to the symptoms it causes and its potential to progress to invasive vulval cancer. Both the International Society for the Study of Vulvovaginal Diseases and the International Society of Gynaecological Pathologists have stressed the importance of eliminating eponymous terminology and recommend only the term vulval intraepithelial neoplasia (VIN). This terminology includes both squamous and non-squamous varieties (Table 1). The latter includes both Paget's disease of the vulva and melanoma in situ. Non squamous VIN will not be considered in this paper. Until 30 years ago VIN was an uncommon condition, seen principally in middle and later life. The incidence particularly in younger women has increased significantly since then [1-3]. Since that time the mean age in our unit has fallen from 52.7 years to 35.8 years (Figure 1) [2]. The increasing incidence of the condition parallels similar trends in cervical intraepithelial neoplasia (CIN) and relates at least in part to changing sexual mores, human papilloma virus (HPV) infection, and cigarette smoking. PMID- 11874068 TI - A ten-year remission maintained by 6,272 mg (3,920 mg/m2) cumulative dose of cisplatin-based chemotherapy for recurrent epithelial ovarian cancer. AB - Since cisplatin is a heavy metal, renal and neurotoxicity is considered to be dose limiting in solid tumors. The current case is unusual in that remission has been maintained in a patient with recurrent epithelial ovarian cancer by cisplatin-based chemotherapy without evidence of renal or neurotoxicity, while receiving a total dose of 6,270 mg (3,920 mg/m2) of cisplatin over 11 1/2 years. PMID- 11874069 TI - Treatment of metastatic invasive moles in two husband-side sisters-in-law. Case reports and review of literature. AB - PURPOSE OF INVESTIGATION: The treatment of "high risk" persistent trophoblastic disease (PTD) consists of poly-chemotherapy. This policy probably will lead to overtreatment of some patients. Also, familiar molar pregnancies through the paternal line are unknown in the literature up till now. METHODS: We describe two cases of "high risk" PTD in two husband-side sisters-in-law, in which poly chemotherapy was stopped after histology became available and showed invasive metastatic mole. CONCLUSION: It should be stressed that treatment decisions should be made based on the concept of "high" or "low" risk PTD, but if histology becomes available, chemotherapy might be less aggressive in cases of invasive mole. If invasive mole could be familiar through the paternal line remains unclear with the current knowledge of genetics in trophoblastic disease. PMID- 11874070 TI - Adjuvant radiotherapy in stage I-II epithelial ovarian cancer. AB - OBJECTIVE: The purpose of this study was to assess the efficacy and side-effects of abdominopelvic irradiation applied as adjuvant postoperative therapy in early stage ovarian carcinomas. METHODS: From 1 January 1988 to 31 December 1993, 113 patients with FIGO stage IA-IIC epithelial ovarian carcinoma were treated with postoperative radiotherapy. Whole abdominal irradiation or lower abdominopelvic irradiation was used. The dose of specification was 20 Gy to the upper part of the abdominal cavity and 40 Gy to the lower part of the abdomen and the pelvic region. RESULTS: Primary cure was achieved in 110 patients (97%). During the period of follow-up, 33 cases of tumor recurrences (30%) were recorded. Abdominopelvic metastases were most frequent (18%). The overall 5-year survival rate for the complete series was 69% and the cancer-specific survival rate was 72%. Tumor grade was an independent and significant prognostic factor (Cox multivariate analysis; p = 0.007). Early radiation reactions of any type were noted in 93% of the cases and, in 11%, discontinuation of radiotherapy was necessary. Late radiation reactions were noted in 58% of the cases and the most common side-effect was diarrhea, but in most cases these reactions were of limited clinical significance. The incidence of severe bowel toxicity was 10% and, in two patients ( 1.8%), surgery was necessary due to late radiation reactions. CONCLUSIONS: Adjuvant abdominopelvic radiotherapy is one option among others (e.g. various types of chemotherapy or no further treatment) in primary treatment of early stage ovarian carcinoma. The optimal adjuvant therapy for this group of patients is not known today and further prospective and randomized studies are needed. PMID- 11874071 TI - Estrogen receptor expression in an endometrial stromal sarcoma after tamoxifen therapy. AB - INTRODUCTION: Several cases of low-grade endometrial stromal sarcomas in women with breast cancer have been reported to be associated with tamoxifen therapy. Estrogen receptor expression has been used to characterize the partial estrogenic action of tamoxifen on the endometrium and has been found in tamoxifen-associated endometrial pathologies. CASE: A low-grade endometrial stromal sarcoma in a woman with a history of breast cancer treated with adjuvant tamoxifen is presented. Steroid receptor studies performed on the tumor were negative for estrogen and positive for progesterone. CONCLUSION: The absence of estrogen receptor expression suggests that endometrial stromal sarcomas are not necessarily caused by the estrogenic properties of tamoxifen. PMID- 11874072 TI - Identifying local tumor variables for operable node-negative, margin-free patients with bulky cervical carcinoma of FIGO stage IB, IIA and IIB without adjuvant therapies. AB - PURPOSE: To identify local risk factors for FIGO IB, IIA and IIB bulky cervical squamous cell carcinoma (tumor size > or = 4 cm) patients with node-negative, margin-free tumors treated by radical hysterectomy, pelvic lymph node and para aortic lymph node dissections without adjuvant therapies. MATERIALS AND METHODS: Thirty-four patients were recruited between 1976 and 1989 because they all declined any postoperative adjuvant therapy. The pathology reports showed that all the specimen margins were free from cancer cells with no para-aortic or pelvic lymph node metastases. The survival interval was calculated starting from the time of surgical intervention to the time of death or the end of this study in the year 2000. RESULT: Tumor variables including cell differentiation, depth of stromal invasion, parametrial invasion, vaginal invasion, uterine body invasion, age, and FIGO stage were analyzed. Only vaginal invasion showed statistical significance for decreasing patient disease-free survival in both univariate and multivariate analyses with p values of 0.003 and 0.002, respectively. CONCLUSION: For node-negative and margin-free patients with bulky cervical squamous cell carcinoma with operable stage IB and IIB, surgical intervention alone could suffice when no vaginal invasion is noted plus an 85% survival rate could be achieved. A prospective pilot study should be initiated although this study showed an excellent survival rate which is perhaps due to the limited number of cases. PMID- 11874073 TI - Piver's radical hysterectomy (type III): Endo-Gia 30 stapler versus traditional forcipressure for resection of the cardinal ligament. AB - OBJECTIVE: Morbidity and costs associated with Piver's radical hysterectomy (type III) are noteworthy. The Endo-Gia stapler method for resection of cardinal ligaments can reduce duration of surgery and hospitalization, blood loss, costs and postoperative infection rates. METHOD: Two groups of patients (homogeneous for age, weight and medical condition) were studied: one group was operated on using the Endo-Gia stapler method (n=52) and the other with the traditional forcipressure (n=13). The size of parametrial tissue removed, blood loss, duration of surgery, duration of hospitalization, cost of materials and postoperative fever were compared in the two groups. RESULT: Mean operative times were lower in the Endo-Stapler group than in the controls (mean 180 min versus 220 min). Mean blood loss was 300 cc in the stapler group versus 450 cc in the forcipressure group. Mean cost of surgery (considering costs of materials, hospital stay. duration of surgery), was lower in the stapler group (3,095 euros) than in the group who underwent traditional surgery (3,434 euros). CONCLUSION: Our data suggest the Endo-Gia stapler method significantly reduces blood loss, operative time and cost. PMID- 11874074 TI - Glutathione S-transferases P1-1 and A1-1 in ovarian cyst fluids. AB - PURPOSE: The purpose of the present study was to determine the gluthathione S transferases (GST) P1-1 and A1-1 levels in cyst fluid from malignant, borderline, and benign ovarian tumors. The clinical relevance of these enzymes in cyst fluid was investigated, including the possible relation with resistance to chemotherapy. METHODS: A total of 90 ovarian cysts were punctured for cyst fluid collection. GSTP1-1 and GSTA1-1 concentrations were determined by ELISA in cyst fluid from 23 malignant, 9 borderline, and 51 benign primary ovarian tumors, and levels were correlated with histopathological data. RESULTS: Significantly higher GSTP1-I concentrations were found in cyst fluid from malignant (median: 477 ng/ml), compared with benign (median: 52 ng/ml) ovarian cysts (p < 0.0001), as well as in fluid from borderline (median: 366 ng/ml) compared with benign cysts (p < 0.0001). No significant differences were found in cyst fluid GSTA1-1 concentrations between the histologic subgroups. In cyst fluid from malignant tumors higher GSTPI-1 and lower GSTAI-1 concentrations were found in patients with worse prognostic factors: FIGO II-III-IV, grade 2-3, residual tumor > 2 cm, presence of ascites, patients with recurrent disease, and survival, but differences were not significant. In the subgroup of patients that received cisplatin-based chemotherapy (n = 14) significantly higher GSTP1-1 (p = 0.01) concentrations were found in patients with recurrence compared with patients without recurrence. Considering only FIGO stage I patients, a differentiation could be made between patients with or without recurrence based on cyst fluid GSTP I - I concentrations. CONCLUSIONS: Determination of glutathione S transferases P 1-1 in cyst fluid samples from ovarian tumors can be of additiona] value in the differentiation between histologic subgroups. In case of possible low malignant potential cysts where sampling of the most representative tissue can be an issue, determination of GSTP- I concentrations in cyst fluid may optimise histopathologic classification. Cyst fluid GSTP1-1 seems to be a good marker for aggressiveness of the ovarian tumor, and it may predict response to chemotherapy. PMID- 11874076 TI - Preoperational diagnosis of a uterine lipoleiomyoma using ultrasound and computed tomography images: a case report. AB - A uterine lipoleiomyoma is a variant of uterine myomas, however, it is rarely found in patients and the diagnosis of uterine lipoleiomyoma has always been in retrospect. Uterine lipoleiomyomas are often diagnosed preoperatively as uterine myomas or ovarian mature teratomas. The key to distinguishing the tumors from lipoleiomyomas is to ascertain the primary site of development--uterus or adnexa. When a large uterine tumor is found in a postmenopausal woman, the possibility of malignancy should be considered. Now, however, advanced modern imaging systems can provide more precise diagnoses than before. The following case illustrates how a uterine lipoleiomyoma was highly suggestive preoperatively based on typical characteristics on ultrasound and computed tomography (CT). A homogeneous hyper echoic mass confined to the uterus on ultrasound initiated the suspiction of the tumor. In addition, the uterine mass showed lower density than water on CT which further established the possibility of a fatty tumor of the uterus. The final pathological examination results confirmed the diagnosis of lipoleiomyoma. PMID- 11874075 TI - Immunohistochemical evaluation of a new epithelial antigen, Ber-EP4, in ovarian cancer: preliminary results. AB - OBJECTIVE: To assess the immunohistochemical expression of Ber-EP4, a new epithelial antigen in ovarian cancer. METHODS: We studied 25 cases of ovarian cancer in which Ber-EP4, CEA and CA 125 were investigated by an immunohistochemical method. We evaluated the correlations between immunohistochemical positivity and grading, histotype and stage of disease. RESULTS: CEA was positive in 5 out of 25 cases (20%), CA-125 in 17 out of 25 cases (68%) and Ber-EP4 in 14 out of 25 cases (56%). Ber-EP4 was mainly present in mucinous tumors in comparison to serous tumors (78.6% vs. 50%). Ber-EP4, as well as CA-125, were directly proportional to tumor differentation (70% of positivity in G1 vs 37.5% in G3 for the former and 80% in G1 vs 50% in G3 for the latter, respectively), whereas CEA showed no relevant difference regarding the grading. There were no differences among the three antigens studied with regard to clinical stage. CONCLUSIONS: In our study Ber-EP4 was positive in 14 out of 22 cases (63.6%) of the primary epithelial ovarian cancers studied. The presence of this antigen seems to be related to histotype and grading but not to clinical stage. PMID- 11874077 TI - Transvaginal ultrasonography and hysterosonography to monitor endometrial effects in tamoxifen-treated patients. AB - PURPOSE OF INVESTIGATION: Our purpose was to evaluate if, during tamoxifen treatment, hysterosonography may increase diagnostic accuracy when compared with transvaginal ultrasonography and to identify, when and in how many cases, further biopsies may be avoided. METHODS: We performed transvaginal utrasound in 310 asymptomatic women under tamoxifen treatment, using 8 mm endometrial thickness as the cut-off. One hundred and seven patients with an endometrium thicker than 8 mm were enrolled for hysterosonography. Parameters to be evaluated by transvaginal ultrasound and hysterosonography were thickness and structural features of the endometrium. It was possible to compare ultrasound examinations with histopathological findings obtained by biopsy in 83 patients. RESULTS: Globally only ten patients from the study cohort had true endometrial pathology. Based on structural features of the endometrium, we found a global accuracy of 95.6%, with 2.8% false negatives and 4.1% false positives. CONCLUSION: Hysterosonography can increase diagnostic accuracy during tamoxifen treatment and may allow further invasive investigations to be avoided in patients with suggestive hysterosonographic features. PMID- 11874078 TI - Bilateral primary squamous cell carcinoma of the ovary: a case report of isolated metastasis to the lateral pelvic wall. AB - Primary squamous cell carcinoma of the ovary is rare. The majority of cases arise most commonly from the lining of a dermoid cyst, and less often in endometriosis or a Brenner tumor. A 40-year-old woman underwent exploratory laparotomy and was found to have a right ovarian tumor adherent to the lateral pelvic wall with no ascites. She underwent total abdominal hysterectomy with bilateral salpingo oophorectomy, pelvic lymphadenectomy, infracolic omentectomy, appendectomy, and right nephrectomy for bilateral primary squamous cell carcinoma of the ovary. She was started on multiagent chemotherapy. On follow-up after two years the patient had died of cerebral metastases. To our knowledge in this report we present the first case in the English literature of bilateral pure squamous cell carcinoma of the ovary. PMID- 11874079 TI - Differential expression of alpha-smooth muscle actin molecule in a subset of bone marrow stromal cells, in b-cell chronic lymphocytic leukemia, autoimmune disorders and normal fetuses. AB - Lymphocytes are a constituent of normal marrow. Both B and T lymphocytes are derived from bone marrow stem cells. Lymphocytes are found in normal marrow as single cells and in lymphoid aggregates or follicles. Lymphocytes and precursors are particularly prominent in bone marrow from children in which they may account for up to 40% of the bone marrow cells. The development of hematopoetic cells within the bone marrow (BM) occurs in intimate association with cells of the bone marrow microenvironment. This phenotypically diverse population of connective tissue-type cells includes fibroblasts, macrophages, adipocytes and endothelial cells and, collectively, represents the stromal tissue of the bone marrow. The presence of myoid cells in human bone marrow has been observed during hemopoiesis in embryonic life, whereas during adult life, it is strictly related to different pathologic conditions such as metastatic carcinoma, Hodgkin's disease, hairy cell leukemia and chronic myelo-proliferative diseases. Under normal circumstances, lymphoid cells may constitute up to 20% of the population of nucleated cells in the bone marrow. However, there may be an absolute or a relative increase, the latter due to a reduction in hematopoietic tissue, as in some skeletal areas in advancing age, or in hypoplastic conditions. The aim of this study was to examine the presence, distribution and quantitation of cells expressing alpha-smooth muscle actin in the stroma of the BM of patients with nodular type b-cell chronic lymphocytic leukemia (B-CLL), patients with autoimmune disorders and embryos (gestational age 15 to 25 weeks). For this reason, we investigated the presence of myoid cells (MCs) in a series of 20 trephine bone marrow biopsies from adult patients and ten fetal specimens of the spine and femur, using a monoclonal antibody recognizing alpha-smooth muscle actin, a contractile microfilament expressed exclusively by smooth muscle cells, myofibroblasts and related cells. The results of our study showed that: 1. BM stromal myoid cells represent a distinct subpopulation of reticular cells in the bone marrow, undergoing cytoskeletal remodeling in response to various stimuli (fetuses). 2. The appearance of BM stromal myoid cells is not only seen as a characteristic feature in B-CLL, but is also seen, to a lesser degree, in the stroma of bone marrow in patients with autoimmune disorders. 3. Stromal cells with phenotypic smooth muscle features appear in bone marrow during pathological situations in a manner reminiscent of what occurs during normal development. PMID- 11874081 TI - Huge primary mucinous cystadenoma of the retroperitoneum mimicking a left ovarian tumor. AB - Primary mucinous cystic tumors of the retroperitoneum are rarely encountered and have been reported in approximately 25 cases in the literature. The histogenesis of primary mucinous cystadenomas is not clear. Most authors suggest that it develops through mucinous metaplasia in a pre-existing mesothelium-lined cyst. Surgery is the only treatment. In this report we present an additional case of primary retroperitoneal mucinous cystadenoma in a 44-year-old female. PMID- 11874080 TI - Prognostic value of p53, c-erb-B2 and MIB-1 in endometrial carcinoma. AB - OBJECTIVE: To assess the immunohistochemical expression of p53 protein, a tumour suppressor gene of the oncogene c-erb-B2 and MIB-1 proliferation marker (Ki-67 antigen) in endometrial carcinoma. METHODS: We studied 29 cases of endometrial carcinoma in which the p53, c-erb-B2 and MIB-1/Ki-67 antigens were investigated by an immunohistochemical method. We evaluated the correlations among the immunohistochemical positivity and the grading, depth of myometrial invasion, stage of the neoplasia and follow-up. RESULTS: Both p53 and c-erb-B2 were positive in 16 out of 29 cases (55.2%), whereas MIB-1 was positive in 19 out of 29 cases (65.5%). All these three antigens showed a positive correlation with the grading, myometrial invasion and FIGO stage. Regarding follow-up, p53, c-erb-B2 and MIB-1 were, respectively, positive in 100%, 83.4% and 66.7% of neoplasias of patients who died of disease whereas they were positive in 40%, 40% and 60%, respectively, of tumours of patients with no evidence of disease. CONCLUSION: The overexpression of p53, c-erb-B2 and MIB-1 seem to indicate a more malignant tumour phenotype. PMID- 11874082 TI - Uterine serous papillary carcinoma clinical and immunopathological study of 9 cases. AB - From January 1993 to December 1998, nine patients with serous papillary endometrial carcinoma (SPEC) were diagnosed and treated at the 2nd Department of Obstetrics and Gynecology, Areteion University Hospital. The incidence of SPEC in our Clinic was 6.77%. The mean age of patients was 65.5 years (range 54-82 years). All patients underwent total abdominal hysterectomy, bilateral salpingo oophorectomy and epiploectomy. Abdominal and para-aortic lymph node sampling was performed in all cases and peritoneal washings were examined cytologically. Histological sections of the specimens, stained with haematoxylin-eosin, were retrieved from the Laboratory of Pathology and re-evaluated by two pathologists. All cases conformed to the diagnostic criteria for SPEC. Immunohistochemical studies were performed in paraffin blocks retrieved from the files, by a streptavidin-viotin method for the detection of vimentin (ENZO monoclonal ab), secretory component (DAKO polyclonal ab), CEA (DAKO monoclonal ab), EMA (DAKO monoclonal ab). The hormonal receptor status, assessed by appropriate positive and negative controls, was studied as well. The presence of mucin and glycogen was studied by histochemical reaction, PAS, PAS diastase and mucicarmine. Serous papillary carcinoma is an unusual but distinct type of endometrical adenocarcinoma, a non-hormonal dependent tumor, with aggressive biologic behavior. Its recognition is mandatory for a correct therapeutic approach. PMID- 11874083 TI - Women with a pelvic mass: indicators of malignancy. AB - OBJECTIVE: To evaluate the efficacy of five methods: pelvic examination (PE), transvaginal ultrasonography (US), Doppler ultrasonography, serum CA125 assay and serum CA72-4 assay, alone or associated, to predict malignancy in patients presenting a pelvic mass originating in the ovary. METHODS: 92 patients underwent a standard protocol for physical examination, CA125, CA72-4, transvaginal ultrasonography and Doppler ultrasonography. RESULTS: Eighteen women were dropped from the study because they had clearly benign masses; two women were dropped from the study because they had clearly malignant lesions. Twenty-two malignant (30%) and 50 benign (70%) pelvic tumors were found. When one method was considered alone the best sensitivity (SENS) was found in physical examination (90%) and the best specificity (SPEC) was found in CA72-4: 88%. If all indicators were positive, the SPEC was 100% but the SENS was 40%. Logistic regression analysis prediction of the character of the pelvic masses was correct in 86%. CONCLUSION: Some additional information to discriminate between malignant and benign pelvic masses can be obtained from the valuation of serum tumor markers, particularly CA72-4. Also Doppler ultrasonography appeared to be useful in the differential diagnosis of pelvic tumors. The prediction of the character of the pelvic masses calculated by a logistic model in which PE, US, CA 125, and CA72-4 are included is very good. PMID- 11874084 TI - Effect of prechemotherapy filgrastim on the bone marrow toxicity of topotecan. AB - PURPOSE: To investigate the efficacy and safety of single-dose filgrastim administered 24 hours prior to chemotherapy in the prevention of topotecan related myeloid suppression. METHODS: No medication was given to 21 rats in group 1; 1.5 mg/m2/day topotecan was administered intraperitoneally for five days every three weeks to 21 rats in group II; a single dose of 5 microg/kg filgrastim was injected intraperitoneally 24 hours before the intraperitoneal administration of the same dose of topotecan to 21 rats in group III. After completion of six cycles of chemotherapy. the rats were decapitated and blood samples were immediately collected into citrated tubes for complete blood counts. RESULTS: White blood cell and lymphocyte counts in the control and the filgrastim + topotecan groups were similar (p > 0.05) and significantly higher than the counts in the topotecan group (p < 0.05). There was no difference in means of neutrophil, monocyte, eosinophil, basophil and erythrocyte counts among the groups (p > 0.05). CONCLUSION: Filgrastim administration prior to chemotherapy seems to be beneficial and further investigations are needed. PMID- 11874086 TI - The role of chemotherapy in malignant mixed mullerian tumors of the female genital tract. AB - Thirteen patients with malignant mixed mullerian tumor of the female genital tract, treated and followed in our clinic from 1989 to 1999 were retrospectively evaluated. Seven patients (53.8%) with advanced disease or postoperative residual tumor were treated with adjuvant chemotherapy. The median age at diagnosis was 64 years (range: 26-79). All patients underwent primary surgical cytoreduction. Tumors were localized to the endometrium in five (62.5%), to the ovaries in two (25%) and to the fallopian tube in one (12.5%) patient. One patient with endometrial carcinosarcoma had a simultaneous second primary ovarian epithelial carcinoma. Two patients (25%) had a heterologous sarcomatous component. Myometrial involvement included less than half the thickness in one patient, while there was no myometrial invasion encountered in two patients. Five patients (38.5%) had more than 50% of the myometrium invaded. Two patients received additional radiotherapy. Six patients received cisplatinum-based chemotherapy (4 had doxorubicin including combinations), while one patient was treated with a doxorubicin+ifosphamide combination. Five patients (71.4%) had a complete response (CR) to chemotherapy. Response duration in patients with a CR was +13, +67, +10, +14 and +2 months, respectively. After a median follow-up period of 20 months (3-115 months), six patients have died, five are being followed-up with no evidence of disease, one is alive with metastatic disease and one patient is under treatment. Malignant mixed mullerian tumor of the female genital tract is highly responsive to multimodality treatment strategies. Further prospective studies are required to identify distinct prognostic groups that may benefit from various treatment modalities. PMID- 11874085 TI - Reproductive tract pathology in asymptomatic women treated with tamoxifen. AB - PURPOSE: To determine the long-term effects of tamoxifen on the female reproductive tract in patients with breast cancer. METHODS: Forty-nine patients with breast cancer receiving tamoxifen longer than two years were analyzed. All the patients underwent pelvic examination, pap smear, transvaginal ultrasonography, serum CA 125 and dilatation and curettage. RESULTS: There were 16 patients with genital system pathology. Three of them had atypical Pap smears, one with cervical carcinoma and the other two with chronic cervicitis. Two significant ovarian pathologies were found. These were ovarian fibroma, and unilateral dermoid cyst. There were three patients with endometrial hyperplasia without atypia. Uterine myoma was encountered in seven of the cases. Only one patient had elevated CA 125 levels despite normal genital examination findings. CONCLUSION: Since no significant genital pathology attributable to tamoxifen therapy could be detected, the follow-up for gynecologic pathologies in breast cancer patients receiving tamoxifen therapy may be individualized. PMID- 11874088 TI - The "push" for evidence: management of the second stage. AB - Recognition that the available evidence does not support arbitrary time limits for the second stage of labor has led to reconsideration of the influence of maternal bearing down efforts on fetal/newborn status as well as on maternal pelvic structural integrity. The evidence that the duration of 'active' pushing is associated with fetal acidosis and denervation injury to maternal perineal musculature has contributed to the delineation of at least two phases during second stage, an early phase of continued fetal descent, and a phase of "active" pushing. The basis for the recommendation that the early phase of passive descent be prolonged and the phase of active pushing shortened by strategies to achieve effective, but non-detrimental pushing efforts is reviewed. The rational includes an emphasis on the obstetric factors that are optimal for birth and conducive to efficient maternal bearing down. Explicit assessment of these obstetric factors and observation of maternal behavior, particularly evidence of an involuntary urge to push, should be coupled with the use of maternal positions that will promote fetal descent as well as reduce maternal pain. The use of epidural analgesia for pain relief can also be accompanied by these same principles, although further research is needed to verify the strategies of "delayed pushing" and maintenance of pain relief along with a reconceptualization of the second stage of labor. PMID- 11874087 TI - Controversies in immunization practices: vaccine safety and implications for midwifery practice. AB - Adverse events occur only rarely after vaccine exposure. Yet, given the high vaccine coverage rates in the United States, the public is increasingly becoming concerned that vaccines may be causing immediate or long-term health problems and less concerned about the possibility of becoming infected. More than 10,000 reports of possible vaccine-related adverse events are reported every year to VAERS, the passive surveillance system that monitors vaccine safety after licensure. As providers of primary health care services to women, midwives are ideally positioned to answer women's questions about vaccine safety. This article provides the background midwives need to be able to help their clients make informed vaccine decisions. It discusses the incidence and risks of infection, the efficacy and risks of vaccines, issues complicating the evaluation of vaccine safety, the state of vaccine safety monitoring systems, and approaches consumers use in vaccine decision making. PMID- 11874089 TI - Late entry into prenatal care in a rural setting. AB - Social support, behavioral risk, and structural or demographic variables as well as acceptance of pregnancy were tested as determinants of late entry into prenatal care in a sample of 176 women in a rural county in California. The respondents were all those over age 18 served by four obstetric practices during a 4-month period ending in February 2000. One nurse-midwifery practice was included. Late entry into prenatal care during the first trimester occurred in 27.3% of the cases overall. Statistically significant independent variables in bivariate analyses were modeled in multivariate logistic regression. Stress, lack of family and friend support. Medicaid enrollment, age under 20 or over 34, low acceptance of pregnancy, and lack of a high school diploma were all predictors of late entry. Lack of family and friend support modified the effects of stress and Medicaid as payer. Although the determinants of late entry were remarkably complex in this sample, they have potential for public health intervention. PMID- 11874090 TI - The 1999-2000 task analysis of American nurse-midwifery/midwifery practice. AB - A master list of tasks, which contained 200 task statements, 23 professional issues statements, and 177 clinical conditions, was divided into three equivalent survey forms and distributed to those certified nurse-midwives (CNMs) and certified midwives (CMs) certified by the ACNM Certification Council, Inc. during the 5-year period from 1995 to 1999. Specific efforts were made to encourage the participation of CMs, because they represented a new professional cohort. A total of 627 valid responses were obtained. Reasonably similar numbers of respondents contributed data related to each of the three versions of the survey form. The responsibilities have expanded substantially within the domains of nonreproductive primary health care and gynecologic care of the well woman, including advances in assisted reproductive technology. A diminished emphasis on the CNM/CM role in the provision of newborn care was documented. The ACC Research Committee recommended the revision of the entry-level certification examination blueprint, and this was approved by the ACC Board of Directors. The specific recommendations included the development of a new primary care domain and the reconfiguration of content emphasis with percentage allocations as follows: Primary Care, 5-10%; Well-Woman/Gynecology, 15-20%; Newborn, 5-10%; Postpartum, 5 10%; Antepartum, 25-30%; Intrapartum, 25-35%; Professional Issues, up to 5%. PMID- 11874091 TI - Gaining ground, teen births in the United States 1940-2000. PMID- 11874092 TI - CNM birth attendance in the United States, 1999. PMID- 11874093 TI - Outcomes of high-risk women cared for by certified nurse-midwives. AB - This study describes the incidence of specific high-risk factors of a population cared for by a group of certified nurse-midwives (CNMs) in a Mid-Atlantic, inner city, nonprofit, hospital-based clinic. Outcomes were compared with all women who delivered in the United States in 1994. Univariate statistics, which consisted of descriptive statistics, frequencies, and percentage distribution, were used. This comparison suggests that CNMs can provide safe care to women with high-risk conditions. Outcomes for the midwifery sample were more favorable for vaginal births, vaginal deliveries after cesarean section, forceps- and vacuum-assisted deliveries, cesarean delivery, and 5-minute Apgar scores. The incidence of maternal fever and meconium stained amniotic fluid was higher. Implications and limitations of the study are discussed. PMID- 11874094 TI - I. Strengthening midwifery in America: reflections from the 46th Annual ACNM Meeting and Exhibit. American College of Nurse-Midwives. PMID- 11874095 TI - II. The multiple art forms within the midwife: reflections from the 46th Annual ACNM Meeting and Exhibit. American College of Nurse-Midwives. PMID- 11874096 TI - Impact of traditional birth attendant training in Mozambique: a controlled study. PMID- 11874097 TI - Early options: educating physicians and women about medical abortion. PMID- 11874098 TI - Expression of the high capacity calcium-binding domain of calreticulin increases bioavailable calcium stores in plants. AB - Modulation of cytosolic calcium levels in both plants and animals is achieved by a system of Ca2+-transport and storage pathways that include Ca2+ buffering proteins in the lumen of intracellular compartments. To date, most research has focused on the role of transporters in regulating cytosolic calcium. We used a reverse genetics approach to modulate calcium stores in the lumen of the endoplasmic reticulum. Our goals were two-fold: to use the low affinity, high capacity Ca2+ binding characteristics of the C-domain of calreticulin to selectively increase Ca2+ storage in the endoplasmic reticulum, and to determine if those alterations affected plant physiological responses to stress. The C domain of calreticulin is a highly acidic region that binds 20-50 moles of Ca2+ per mole of protein and has been shown to be the major site of Ca2+ storage within the endoplasmic reticulum of plant cells. A 377-bp fragment encoding the C domain and ER retention signal from the maize calreticulin gene was fused to a gene for the green fluorescent protein and expressed in Arabidopsis under the control of a heat shock promoter. Following induction on normal medium, the C domain transformants showed delayed loss of chlorophyll after transfer to calcium depleted medium when compared to seedlings transformed with green fluorescent protein alone. Total calcium measurements showed a 9-35% increase for induced C domain transformants compared to controls. The data suggest that ectopic expression of the calreticulin C-domain increases Ca2+ stores, and that this Ca2+ reserve can be used by the plant in times of stress. PMID- 11874099 TI - Expression of a synthetic porcine alpha-lactalbumin gene in the kernels of transgenic maize. AB - The main nutritional limitation of maize used for feed is the content of protein that is digestible, bioavailable and contains an amino acid balance that matches the requirements of animals. In contrast, milk protein has good digestibility, bioavailability and amino acid balance. As an initial effort to create maize optimized as a source of swine nutrition, a codon-adjusted version of a gene encoding the milk protein porcine alpha-lactalbumin was synthesized. Maize expression vectors containing this gene under the control of the Ubi-1 promoter and nos 3' terminator were constructed. These vectors were used to transform maize callus lines that were regenerated into fertile plants. The alpha lactalbumin transgenes were transmitted through meiosis to the sexual progeny of the regenerated plants. Porcine alpha-lactalbumin was detected in callus and kernels from transgenic maize lines that were transformed by two constructs containing the 27-kDa maize gamma-zein signal sequence at the 5' end of the synthetic porcine alpha-lactalbumin coding sequence. One of these constructs contained an ER retention signal and the other did not. Expression was not observed in kernels or callus from transgenic maize lines that were transformed by a construct that does not contain an exogenous protein-targeting signal. This suggests that the signal peptide might play an important role in porcine alpha lactalbumin accumulation in transgenic maize kernels. PMID- 11874101 TI - Resistance to wheat streak mosaic virus in transgenic wheat engineered with the viral coat protein gene. AB - Wheat (Triticum aestivum) plants were stably transformed with the coat protein (CP) gene of wheat streak mosaic virus (WSMV) by the biolistic method. Eleven independently transformed plant lines were obtained and five were analyzed for gene expression and resistance to WSMV. One line showed high resistance to inoculations of two WSMV strains. This line had milder symptoms and lower virus titer than control plants after inoculation. After infection, new growth did not show symptoms. The observed resistance was similar to the 'recovery' type resistance described previously using WSMV NIb transgene and in other systems. This line looked morphologically normal but had an unusually high transgene copy number (approximately 90 copies per 2C homozygous genome). Northern hybridization analysis indicated a high level of degraded CP mRNA expression. However, no coat protein expression was detected. PMID- 11874100 TI - Conditional gene expression in the respiratory epithelium of the mouse. AB - Transgenic mouse models mediating conditional temporal and spatial regulation of gene expression to the respiratory epithelium were developed utilizing the reverse tetracycline transactivator (rtTA) expressed under the control of SP-C and CCSP promoters. Luciferase activity was detected in the lungs of fetal and adult double transgenic mice but was not detected in other tissues or in single transgenic mice. In adult mice, maximal luciferase activity was detected 16 h after the administration of doxycycline in the drinking water, or 2 h after the injection of doxycycline. Activation of the transgene was observed after the administration of doxycycline in food pellets. After prolonged exposure to doxycycline, luciferase activity decreased slowly following removal of doxycycline, suggesting the importance of tissue pools which maintained expression of the transgene. In SP-C-rtTA mice, exposure of the pregnant dam to doxycycline induced luciferase activity in fetal lung tissue as early as E10.5. Luciferase activity was maintained in the lung tissue of pups during the period of lactation when the mother received doxycycline in the drinking water. In the CCSP-rtTA mice, luciferase was not detected in the absence of doxycycline. In the SP-C-rtTA mice, luciferase activity was detected in the absence of doxycycline but was enhanced approximately 10-fold by administration of drugs. The SP-C-rtTA and CCSP-rtTA activator mice control the expression of transgenes in the developing and mature respiratory epithelium, and will be useful for the study of gene function in the lung. PMID- 11874102 TI - Rapid and accurate determination of zygosity in transgenic animals by real-time quantitative PCR. AB - Successful identification of homozygous and heterozygous transgenic animals with currently available techniques demands tedious and time-consuming procedures with a high proportion of ambiguous results. Real-time PCR is a quantitative and extremely precise method with high throughput that could be applied to the analysis of large numbers of animals differing only by a factor of two in the amount of target sequences. We defined the technical conditions of real-time PCR to co-amplify a transgene and a reference gene using two fluorogenic probes and the comparative cycle threshold method. We applied these conditions to the analysis of zygosity in a line of transgenic rats. Real-time PCR allowed clear cut identification of all transgenic animals analysed (n = 45) as homozygous or heterozygous. Southern blot analysis of these animals using an internal quantitative control and PhosphorImager quantification showed ambiguous results in six of them and was concordant with real-time PCR in the rest. Mating of homozygous and heterozygous animals, as defined by real-time PCR, showed transgene transmission to the offspring following expected Mendelian laws. Real time PCR allows rapid, precise, non-ambiguous and high throughput identification of zygosity in transgenic animals. This technique could be helpful in the establishment of breeding programs for transgenic colonies and in experiments in which gene dosage effects could have a functional impact. PMID- 11874103 TI - Expression of a single-chain Fv antibody fragment specific for the hepatitis B surface antigen in transgenic tobacco plants. AB - An anti-Hepatitis B virus surface antigen (HBsAg) single chain Fv (scFv) antibody fragment was expressed in Nicotiana tabacum transgenic plants. The 6-histidine tagged scFv was targeted to either the cytosol, apoplast, and vacuole, or for retention in the endoplasmic reticulum. Expression of active scFv was detected by ELISA in fresh leaf material from F I transgenic plant lines representative of the genetic constructs targeting the antibody fragment to the apoplastic fluid (AF-12, 0.031% of the total soluble protein), vacuole (V-20, 0.032% of the total soluble protein), and endoplasmic reticulum (ER-52, 0.22% of the total soluble protein). No scFv was detected by ELISA or western blot in the plants transformed with the cytosol construct. The biologically active scFv was easily purified (to 94-95% purity) from ER-52 and AF-12 plant material using immobilized metal ion affinity chromatography. Recovery estimated from the ER-52 plant line indicates that 15-20 microg of pure active scFv can be obtained per gram of fresh leaf material, on a laboratory scale. PMID- 11874104 TI - Ubiquitous expression of goat cyclin T1 in transgenic mice. AB - Ubiquitous gene expression has a variety of applications in transgenesis that include, for example cell lineage analyses in chimeras and gain-of-function related to xenotransplantations. Although several promoters have already been used to these aims, they often do not reliably or reproducibly target gene expression in mice. We have recently reported the site-independent expression of a bacterial artificial chromosome (BAC)-derived goat alpha-lactalbumin transgene in the mammary gland of mice and the subsequent localisation within the insert of this BAC of the cyclin T1 locus. This ubiquitously expressed gene encodes for a protein that acts as a co-factor for the HIV nuclear transcriptional activator. In the present paper, we report that the goat BAC transgene, which encompasses around 30 kb of the cyclin T1 promoter, also confers ubiquitous expression of this gene in the six transgenic mouse lines analysed. These results suggest that the cyclin T1 promoter could be a useful alternative to target ubiquitous gene expression in transgenics. PMID- 11874105 TI - Transformation vector based on promoter and intron sequences of a replacement histone H3 gene. A tool for high, constitutive gene expression in plants. AB - This study explored the possibility of using non-viral, plant-based gene sequences to create strong and constitutive expression vectors. Replacement histone H3 genes are highly and constitutively expressed in all plants. Sequences of the cloned alfalfa histone H3.2 gene MsH3gl were tested. Constructs of the beta-glucuronidase (GUS) reporter gene were produced with H3.2 gene promoter and intron sequences. Their efficiency was compared with that of the commonly used strong 35S cauliflower mosaic virus promoter in transgenic tobacco plants. Combination of the H3.2 promoter and intron produced significantly higher GUS expression than the strong viral 35S promoter. Histochemical GUS analysis revealed a constitutive pattern of expression. Thus, alfalfa replacement H3 gene sequences can be used instead of viral promoters to drive heterologous gene expression in plants, avoiding perceived risks of viral sequences. PMID- 11874106 TI - No credible scientific evidence is presented to support claims that transgenic DNA was introgressed into traditional maize landraces in Oaxaca, Mexico. PMID- 11874107 TI - A single-cell matrix-assisted laser desorption/ionization time-of-flight mass spectroscopic assay of the cell-maturation process. PMID- 11874108 TI - Using reactands in CMOS-based calorimetric sensors: new functional materials for electronic noses. PMID- 11874109 TI - Highly selective transport of a uranyl ion through a liquid membrane containing a lipophilic ion-associate of methyltrioctylammonium and hydroxycalix[n]arene-p sulfonates (n = 6, 8) as metal carriers. AB - Uranyl ion, UO2(2+), in an aqueous sodium hydrogen carbonate solution of pH 4-8 (source phase) was simultaneously and selectively transported into a dilute sulfuric acid solution (receiving phase) through a membrane (chloroform, bulk) containing a lipophilic ion-associate of methyltrioctylammonium ion and hydroxycalix[n]arene-p-sulfonate ion, 2n (n = 6, 8), MTA+-2n, as a metal carrier. The rate of transport increased in proportion to the concentrations of UO2(2+) in the source phase and carrier in the membrane and along with an increase in the temperature of the system. The rate was also increased along with an increase in the pH of the source phase. None of the other metal ions were transported, or obstructed the transport of UO2(2+), while the presence of large amounts of sodium hydrogencarbonate and sodium chloride in the source phase interfered with the transport by causing a delay in the start of transport. PMID- 11874110 TI - Guest-responsive fluorescence variations of gamma-cyclodextrins labeled with hetero-functionalized pyrene and tosyl moieties. AB - Regioselectively hetero-labeled hosts, 6A-pyrenebutylate-6X-tosyl-modified gamma cyclodextrins (X = B or H, C or G, D or F, and E for gamma-1, gamma-2, gamma-3, and gamma-4, respectively), were synthesized in order to investigate their chemo sensor properties for applications to organic compounds, such as bile acids and terpenes. The hosts (gamma-1, gamma-2, gamma-3, and gamma-4) exhibit pure monomer fluorescence. The guest-induced fluorescence emissions of these hosts were suppressed in the presence of guests. The extent of fluorescence variations of these hosts with guests was recognized as a manifestation of the sensing ability of the hosts. A sensing parameter (deltaI/I0, where I and I0 are the fluorescence intensities in the presence and absence of a guest and deltaI = I0-I) was used to describe the sensing ability of these hosts. Host gamma-analogs were able to detect progesterone, ursodeoxycholic acid, chenodeoxycholic acid, and (-)-borneol with high sensitivity. The behaviors of the appended moieties of these hosts during the formation of host-guest complexes were studied using induced circular dichroism (ICD) spectra, fluorescence spectra and the MM2-energy-minimized structure. The host gamma-analogs exhibited different ICD spectra patterns before and after the addition of ursodeoxycholic acid. The guest-induced variations of ICD and the fluorescence spectra and MM2-minimized structures suggest that the pyrene and tosyl moieties move by altering the spatial relationship between them, in which the pyrene moiety works as a hydrophobic cap and the tosyl moiety is speculated to act as a spacer. PMID- 11874111 TI - Novel spectrophotometric method for the determination of molybdenum in a PVA medium. AB - A novel spectrophotometric method for the determination of trace molybdenum in the presence of a large amount of tungsten was developed. This proposed method was based on the formation of a green charge-transfer polyoxometalate, molybdo-11 tungstophosphate-3,3',5,5'-tetramethylbenzidine-N-propanesulfonic, which was solubilized and stabilized in a PVA medium; the wavelength of the maximum absorption was at 660 nm. Beer's law was obeyed in the Mo concentration range of 0.04-1.6 microg ml(-1). The molar absorptivity was 1.35 x 10(4) l mol(-1) cm(-1). The developed method avoided the interference of W(VI), and was conveniently applied to analyses of some tungsten ores containing Mo(VI) with satisfactory results. PMID- 11874112 TI - Nitric oxide biosensors based on Hb/phosphatidylcholine films. AB - Hemoglobin (Hb) was entrapped in a phosphatidylcholine (PC) film and immobilized at a pyrolytic graphite (PG) electrode surface. Its electron-transfer reactivity and enzyme activity were characterized by employing electrochemical methods. It was observed that Hb exhibited direct electrochemistry as well as enzyme-like activity towards the electrocatalytic reduction of NO in PC film. An unmediated, reagentless nitrogen oxide (NO) biosensor was accordingly prepared. Experimental results revealed that the peak current related to NO was linearly proportional to its concentration in the range of 1.0 x 10(-7)-3.0 x 10(-4) mol/L. The detection limit was estimated to be 1.0 x 10(-7) mol/L. Considering its good stability, nice selectivity and easy construction, this biosensor shows great promise for the rapid determination of traces of NO. PMID- 11874113 TI - Clotrimazole-triiodide ion association as an ion exchanger for a triiodide ion selective electrode. AB - A novel triiodide ion-selective electrode based on a clotrimazole-triiodide ion pair as a membrane carrier was prepared. It has a linear response to triiodide from 8 x 10(-6) to 5 x 10(-3) M with a slope of -68.9 mV per decade and a detection limit of 5 x 10(-6) M. The electrode response is independent of the pH of the solution in the pH range 2-9. It has a very short response time and can be used for at least 3 months without any considerable divergence in the potentials. The proposed sensor revealed very good selectivities for I3- over a variety of other anions. It was used as an indicator electrode in the potentiometric titration of triiodide ions and in an indirect potentiometric determination of clotrimazole in pharmaceutical preparations. PMID- 11874114 TI - Design of a new dodecyl sulfate-selective electrode based on conductive polyaniline. AB - A new, simple, sensitive, low cost and rapid potentiometric method for direct determination of ultra trace amounts of sodium dodecyl sulfate (SDS) with a new DS(-)-selective electrode is reported. The electrode was prepared by electropolymerization of aniline in acidified DS- ion on the surface of a Pt electrode. The cyclic voltammetry (CV) was used for electropolymerization of polyaniline (PA) in the potential range of -200 to +1000 mV vs. Ag/AgCl. This sensor showed a Nernstian behavior (59.0 +/- 2.3 mV/decade) over a very wide linear range (1.0 x 10(-9)-3.0 x 10(-6) M) with a detection limit of 1.0 x 10(-9) M. The response time of the electrode was 15 s for 1.0 x 10(-7) M of analyte; the electrode can be used for 4 weeks without any major deviation. This electrode can be used in the pH range of 3.5-9.8. The selectivity of electrode to DS- over some organic, inorganic and anionic surfactants was investigated with the fixed primary ion method. The results show that the electrode is highly selective to DS ion over other ions. The proposed electrode was applied to the determination of DS- in real samples. PMID- 11874115 TI - Component analysis of the commercial metal extractant, Cyanex 302, by GC-MS. AB - The composition of the commercial reagent Cyanex 302 was investigated by GC-MS. Mass spectrometric studies allowed us to confirm that bis(2,4,4 trimethylpentyl)monothiophosphinic acid is the major compound and that a considerable amount of tris(2,4,4-trimethylpentyl)phosphine oxide is also present. The study also revealed that the extractant has three minor components. These were identified as bis(2,4,4-trimethylpentyl)phosphinic acid, bis(2,4,4 trimethylpentyl)phosphine oxide and bis(2,4,4-trimethylpentyl)dithiophosphinic acid. The mass spectra of these compounds are discussed and some fragmentation processes are postulated. PMID- 11874116 TI - A novel two-enzyme amperometric electrode for lactose determination. AB - Coimmobilization of beta-galactosidase and glucose oxidase in a redox polymer, polyvinylferrocenium perchlorate (PVF+ ClO4-), led to the development of an enzyme electrode for the determination of lactose. The amperometric response of the electrode was measured at +0.70 V vs. SCE, which was due to the electrooxidation of enzymatically produced H2O2. The effects of the substrate and buffer concentrations as well as the pH on the electrode response were elucidated. PMID- 11874117 TI - Retention of anions on silica-based metalloporphyrin stationary phases. AB - The silica-based Fe(III)-protoporphyrin and Zn-tetraphenylporphyrin stationary phases were examined for the HPLC separation of anions. The retention of nine common inorganic anions as well as benzoate anion (BA) and its hydroxy analogues (HBA) was examined using tartrate, acetate, and succinate eluents. The retention factors of inorganic anions on the FeProP stationary phase were in the order Cl- < NO3- < ClO4- < I- < SCN- and for organic anions benzoate < p-hydroxybenzoate < m-hydoxybenzoate < o-hydroxybenzoate. The retention factors of organic anions examined for a ZnTPP column were in the order p-HBA < m-HBA < BA < o-HBA. PMID- 11874118 TI - An amperometric immunosensor based on a conducting immunocomposite electrode for the determination of Schistosoma japonicum antigen. AB - A renewable amperometric immunosensor based on a graphite-paraffin-Schistosoma japonicum antibody (SjAb) biocomposite electrode has been prepared for the detection of Schistosoma japonicum antigen (SjAg). Competitive ELISA was employed involving HRP-SjAg as a tracer and 3,3',5,5'-tetramethylbenzidine (TMB) as a substrate. The product of an enzyme catalytic reaction was detected at +0.1 V (vs. Ag/AgCl reference electrode) for measuring the amount of HRP-labeled SjAg binding to the electrode surface. The assay conditions were optimized, including the amount of SjAb loading in the electrode and HRP-SjAg in the incubation solution, the pH of the measuring solution and the incubation time. The measuring range was 0.5-30 microg/ml under the optimum conditions. Rabbit serum samples of different infection degree were measured, which demonstrated that the immunosensor meets the demands of clinical analysis. It exhibits some advantages, such as simplicity of fabrication, rapidity of measurement, and satisfactory sensitivity and reproducibility. PMID- 11874119 TI - Syntheses and characterizations of 4-(3,17beta-dihydroxyestra-1,3,5(10)-trien 6alpha- and 6beta-yl)amino-7-nitro-2,1,3-benzoxadiazoles as fluorescent probes. AB - 4-(3,17Beta-dihydroxyestra-1,3,5(10)-trien-6alpha- and 6beta-yl)amino-7-nitro 2,1,3-benzoxadiazoles have been synthesized and characterized as fluorescent probes for use in a receptor assay and/or a homogeneous immunoassay for estradiol. The fluorescence intensities are strongly dependent upon the solvent polarity used. The intensities in water were reduced to less than 1% of those in ethyl acetate, and a blue shift was also observed in polar solvents. The quenched fluorescence in aqueous solution was recovered by adding bovine serum albumin or an anti-estradiol antibody. Adding intact estradiol inhibited the fluorescence recovered by the antibody. PMID- 11874120 TI - Spectrophotometric determination of cerium(IV) using a phenothiazine derivative. AB - A simple, rapid and sensitive spectrophotometric method has been proposed for the determination of cerium(IV) using a phenothiazine derivative, propionyl promazine phosphate (PPP). This method is based on the formation of a red-colored radical cation upon a reaction of PPP with cerium(IV) in a phosphoric acid medium having maximum absorbance at 513 nm. Beer's law is valid over the concentration range of 1-11 microg/ml with a Sandell's sensitivity value of 16.14 ng/cm2. The proposed method has been successfully applied to the analysis of magnesium-base cerium alloys and synthetic mixtures corresponding to various cerium alloys. Other phenothiazine derivatives viz. butaperazine dimaleate and propericiazine were also used for the determination of cerium(IV). PMID- 11874121 TI - Analysis of residual solvents in ampicillin powder by headspace spectrophotometric method. AB - In this study a headspace spectrophotometric method is proposed for analysis of dichloromethane and isobutyl methyl keton (IBMK) residues in the ampicillin powder. Ampicillin is dissolved in 1 M NaOH in the vessel of an arsenic analyzer unit of an atomic absorption spectrophotometer. After 3-min stirring, the headspace has flowed by air into the flow-through cell and its absorbance is read at 196 nm, as emitted by a selenium hollow cathode lamp. The absorbance of the headspace is read in two cases (in the presence and absence of MnO4- ion). In the former case, the absorbance is only related to dichloromethane; in the latter, it is related to both solvents. By this method both solvents are determined in the ampicillin samples. The obtained results are compared with gas chromatography (GC) data. These results have good agreement. The proposed method is very rapid, selective and repeatable. Other solvents present, such as isopropyl alcohol, ethylacetate and triethylamine, are not interfering. PMID- 11874122 TI - A resonance light-scattering determination of proteins with fast green FCF. AB - The interaction of Fast Green FCF (FCF) with proteins (including bovine serum albumin (BSA), human serum albumin (HSA), pepsin (Pep) and alpha-chymotrypsin (Chy), and lysozyme (Lys)) was characterized by enhanced resonance light scattering (RLS) measurements using a common spectrofluorometer. The enhanced RLS signals of FCF by proteins at 279.0 nm were obtained, and the mechanism of the RLS enhancement was considered in terms of the effects of the pH and ionic strength on the interaction. It was found that the enhanced RLS intensities were in proportion to the concentrations of proteins in the range of nanogram levels, displaying that the present assay is much more sensitive than the reported RLS methods, with the limits of determination being 4.54, 0.6, 22.8, 4.32 and 1.75 ng/ml for BSA, HSA, Pep, Chy, and Lys. respectively. PMID- 11874123 TI - Square-wave adsorptive stripping voltammetric determination of the anti inflammatory drug lornoxicam. PMID- 11874124 TI - Determination of mercury at a dithizone-modified glassy carbon electrode by anodic stripping voltammetry. PMID- 11874125 TI - Simultaneous determination of nickel and cadmium by differential pulse polarography. PMID- 11874126 TI - Accumulation voltammetry of MoO4(2-) at a glassy carbon electrode covered with chitin film. PMID- 11874127 TI - Alumina and admicellar sorbents for the separation of copper(II) ions and humic complexes in water. PMID- 11874128 TI - Preparation of poly(N-isopropylacrylamide)-modified glass surface for flow control in microfluidics. PMID- 11874129 TI - The influence of triton X-405 on the fluorescence properties of abscisic acid. PMID- 11874130 TI - Crystal structure of 4-[[(1E)-(2-hydroxynaphthyl)methylidene]amino]-1,5-dimethyl 2-phenyl-2,3-dihydro-1H-pyrazol-3-one. PMID- 11874131 TI - Crystal structure of 2alpha,3alpha-dihydroxycativic acid: 6,7-dihydroxy 1,4,4a,5,6,7,8,8a-octahydro-beta,2,5,5,8a-pentamethyl-1-naphthalene pentanoic acid. PMID- 11874132 TI - Crystal structure of 2,5-bis-(3,5-dimethylbenzyl)-3,6-dinaphthalen-2-yl-2,5 dihydro-pyrrolo[3,4-c]pyrrole-1,4-dione. PMID- 11874133 TI - Crystal structure of 2,5-bis-(3,5-di-tert-butyl-benzyl)-3,6-bis-(4-dimethylamino phenyl)-2,5-dihydro-pyrrolo[3,4-c]pyrrole-1,4-dione. PMID- 11874134 TI - The effects of prolonged intracortical microstimulation on the excitability of pyramidal tract neurons in the cat. AB - This study was conducted to examine the excitability changes induced in cerebral cortical neurons during prolonged microstimulation with a spatially dense microelectrodes array. The arrays of 16 iridium microelectrodes were implanted chronically into the postcruciate gyrus of cats. Neuronal responses characteristic of single pyramidal tract axons (ULRs) were recorded in the medullary pyramid. 7 h of pulsing of individual electrodes at 50 Hz and at 4 nC/ph induced little or no change in the ULRs' electrical thresholds. The thresholds also were quite stable when 4 of the 16 microelectrodes were pulsed on each of 14 consecutive days. However, when all 16 microelectrodes were pulsed for 7 h at 4 nC/ph, the threshold of approximately half of the ULRs became elevated. Recovery of excitability required 2-18 days. Prolonged sequential (interleaved) pulsing of the 16 microelectrodes induced less depression of excitability than did simultaneous pulsing, but only when the stimulus amplitude was low (12 A, 1.8 nC/ph). Stimulation at a higher amplitude (15 nC/ph) induced much more depression of excitability. These findings imply that multiple processes mediate the stimulation-induced depression of neuronal excitability. The data also demonstrate that the depression can be reduced by employing a stimulus regimen in which the inherent spatial resolution of the array is maximized (sequential pulsing at an amplitude in which there is minimal overlap of the effective current fields). PMID- 11874135 TI - Ultrafast flow quantification with segmented k-space magnetic resonance phase velocity mapping. AB - Magnetic resonance (MR) phase-velocity mapping (PVM) is routinely being used clinically to measure blood flow velocity. Conventional nonsegmented PVM is accurate but relatively slow (3-5 min per measurement). Ultrafast k-space segmented PVM offers much shorter acquisitions (on the order of seconds instead of minutes). The aim of this study was to evaluate the accuracy of segmented PVM in quantifying flow from through-plane velocity measurements. Experiments were performed using four straight tubes (inner diameter of 5.6-26.2 mm), under a variety of steady (1.7-200 ml/s) and pulsatile (6-90 ml/cycle) flow conditions. Two different segmented PVM schemes were tested, one with five k-space lines per segment and one with nine lines per segment. Results showed that both segmented sequences provided very accurate flow quantification (errors<5%) under both steady and pulsatile flow conditions, even under turbulent flow conditions. This agreement was confirmed via regression analysis. Further statistical analysis comparing the flow data from the segmented PVM techniques with (i) the data from the nonsegmented technique and (ii) the true flow values showed no significant difference (all p values>>0.05). Preliminary flow measurements in the ascending aorta of two human subjects using the nonsegmented sequence and the segmented sequence with nine lines per segment showed very close agreement. The results of this study suggest that ultrafast PVM has great potential to measure blood velocity and quantify blood flow clinically. PMID- 11874136 TI - Parallel cascade recognition of exon and intron DNA sequences. AB - Many of the current procedures for detecting coding regions on human DNA sequences combine a number of individual techniques such as discriminant analysis and neural net methods. Recent papers have used techniques from nonlinear systems identification, in particular, parallel cascade identification (PCI), as one means for classifying protein sequences into their structure/function groups. In the present paper, PCI is used in a pilot study to distinguish exon (coding) from intron (noncoding; interspersed within genes) human DNA sequences. Only the first exon and first intron sequences with known boundaries in genomic DNA from the beta T-cell receptor locus were used for training. Then, the parallel cascade classifiers were able to achieve classification rates of about 89% on novel sequences in a test set, and averaged about 82% when results of a blind test were included. In testing over a much wider range of human nucleotide sequences, PCI classifiers averaged 83.6% correct classifications. These results indicate that parallel cascade classifiers may be useful components in future coding region detection programs. PMID- 11874137 TI - Assay procedure optimization of a rapid, reusable protein C immunosensor for physiological samples. AB - A fiber-optic immunosensor for quantifying the protein C (PC) amount in a plasma sample is being developed to provide accurate, fast, cost-effective diagnosis of heterozygous PC deficiency (0.5-2.5 microg ml(-1)). As a progress report on the sensor development, this paper focuses on optimizations of vanous assay steps. These include: (1) the primary antibody concentration; (2) the effect of the primary antibody leaching on the sensitivity; (3) the fluorophore to secondary antibody ratio; and (4) the sample and secondary antibody incubation times. The optimal primary antibody concentration for the PC sensor was determined to be 65 microg ml(-1); its leaching was minor, stabilized within 3 days, and the sensor sensitivity change after 30 days of storage was minor; sample and secondary antibody incubation times were reduced from 10 to 5 and from 5 to 3 min, respectively, while maintaining a reasonable signal-to-noise level. The immunosensor was also tested with (5) human serum albumin and human plasma and (6) with and without convective flow. High viscosity of human plasma decreased signal intensity, but clear signal discrimination was possible in the concentration range of interest. (7) Application of convective flow increased the signal intensity by increasing PC mass transport rate to the sensor surface. The optimized PC immunosensor provides a smaller (100 microl sample chamber) and faster (10-15 min) tool for PC detection in human plasma. PMID- 11874139 TI - The National Institute of Biomedical Imaging and Bioengineering: history, status, and potential impact. AB - This paper describes the history, current status, and objectives and potential impact of the new National Institute of Biomedical Imaging and Bioengineering (NIBIB). Three of the authors (Hendee, Chien, and Maynard) have been involved over several years in the effort to raise the identity of biomedical imaging and bioengineering at the National Institutes of Health. The fourth author (Dean) is the Acting Director of the newly formed NIBIB. These individuals have an extensive collective knowledge of the events that led to formation of the NIBIB, and are intimately involved in shaping its objectives and implementation strategy. This special report provides a historical record of activities leading to establishment of the NIBIB, and an accounting of present and potential advances in biomedical engineering and imaging that will be facilitated and enhanced by NIBIB. The National Institute of Biomedical Imaging and Bioengineering represents a "coming of age" of biomedical engineering and imaging, and offers great potential to expand the research frontiers of these disciplines to unparalleled heights. PMID- 11874138 TI - Methodology for estimation of time-dependent surface heat flux due to cryogen spray cooling. AB - Cryogen spray cooling (CSC) is an effective technique to protect the epidermis during cutaneous laser therapies. Spraying a cryogen onto the skin surface creates a time-varying heat flux, effectively cooling the skin during and following the cryogen spurt. In previous studies mathematical models were developed to predict the human skin temperature profiles during the cryogen spraying time. However, no studies have accounted for the additional cooling due to residual cryogen left on the skin surface following the spurt termination. We formulate and solve an inverse heat conduction (IHC) problem to predict the time varying surface heat flux both during and following a cryogen spurt. The IHC formulation uses measured temperature profiles from within a medium to estimate the surface heat flux. We implement a one-dimensional sequential function specification method (SFSM) to estimate the surface heat flux from internal temperatures measured within an in vitro model in response to a cryogen spurt. Solution accuracy and experimental errors are examined using simulated temperature data. Heat flux following spurt termination appears substantial; however, it is less than that during the spraying time. The estimated time varying heat flux can subsequently be used in forward heat conduction models to estimate temperature profiles in skin during and following a cryogen spurt and predict appropriate timing for onset of the laser pulse. PMID- 11874140 TI - The concept of using multifrequency energy transmission to reduce hot spots during deep-body hyperthermia. AB - We propose a multifrequency energy transmission scheme for annular phased arrays in deep-body hyperthermia. The problem of frequency dependent resonance effects reported in hyperthermia phantom calibration studies, leading to skewed energy deposition patterns and unwanted energy maxima (hot spots), is addressed by comparing broadband energy transmission with conventional monofrequency excitation. A heating system, including microstrip spiral applicators placed at a cylindrical surface in four quadrants, was designed to study this new concept in a homogeneous phantom. The system includes an option to select three cw tones for simultaneous transmission within a frequency range from 232 to 936 MHz. Quantitative analyzes of E-field probe-scan data, generated in a 6 g/L saline phantom and evaluated by three figures of merits defined, demonstrate the possibility of adding up power in the focus site while smearing secondary maxima outside this volume. Random selection of the three sinusoids within this band shows a probability of 0.7 to improve performance compared to the conventional single-frequency heating. A numerical study of the electromagnetic problem was also implemented using the FD-TD method. Analysis shows acceptable agreement between measured and simulated two-dimensional energy deposition patterns. PMID- 11874141 TI - Using uniaxial pseudorandom stress stimuli to develop soft tissue constitutive equations. AB - A nonlinear systems identification method was used to develop constitutive equations for soft tissue specimens under uniaxial tension. The constitutive equations are developed from a single test by applying a pseudorandom Gaussian (PGN) stress input to the specimen, measuring the resulting strain, and calculating the Volterra-Wiener kernels. First and second order kernels were developed for two tissues with widely different properties, rat medial collateral knee ligaments, and rat skin. These kernels were used to predict the strain response to a variety of sinusoidal stress inputs. These predicted strains were compared with the measured strain response using the normalized mean squared error (NMSE). Results showed NMSEs in the range of 0.01-0.08 provided that the magnitudes of the applied stresses were present in the original PGN stress input. Overall, the method provides a means to develop soft tissue constitutive equations that can predict both nonlinear and viscoelastic behavior over a wide range of stress inputs. PMID- 11874142 TI - Neural and mechanical contributions to the stretch reflex: a model synthesis. AB - A model for the soleus stretch reflex in the decerebrate cat was synthesized from models of the neural and muscular components, including the two proprioceptors (the muscle spindle and Golgi tendon organ) and their associated afferents (Ia, II, and Ib), the alpha motoneuron pool with its reflex pathways, the branches of the alpha motoneurons to the intrafusal muscles (beta innervation), and the extrafusal muscle. Parameters for the muscle and receptor models were chosen independently to match their responses in isolation. Reflex gains and gamma inputs were estimated to fit the response to stretch measured by Nichols and Houk. The chosen reflex gains and gamma inputs are not unique; many different combinations reproduced the characteristic stretch response. With a single set of fixed parameters, the model predicted many mechanical properties of the stretch reflex, including linearization effects (when the stretch magnitude and direction are varied), as well as the dependence on operating force and initial muscle length. The model did not accurately predict the responses at higher stretch velocities, due to failure of the extrafusal muscle model. PMID- 11874143 TI - Structural model of the muscle spindle. AB - A model of the muscle spindle was developed based on its anatomical structure. The model contains three intrafusal fibers (bag1, bag2, and chain), two efferents (dynamic gamma efferent to the bag1 fiber and static gamma efferent to bag2 and chain fibers), and two afferents [primary (Ia) and secondary (II)]. As in the real muscle spindle, the spindle model, under the modulation of gamma efferents, responds to the extrafusal muscle fiber length. Both outputs (Ia and II afferents) of the model were compared extensively with published data, under both sinusoidal stretch (with different stretch amplitudes and frequencies) and ramp and hold stretch (with different stretch amplitudes and velocities) in three different fusimotor activation conditions (dynamic gamma stimulation, static gamma stimulation, and without gamma stimulation). Model Ia afferent responses fit the published data well with active gamma input, but less well in the passive state. Model II afferent responses also fit the published data, although less quantitative data were available for comparison. The model correctly predicted the fractional power dependence of the primary and secondary ending responses on stretch velocity. The current model provides a powerful tool for simulation studies of neuromusculoskeletal systems, and demonstrates the feasibility of using a structural approach to model complex neurophysiological systems. PMID- 11874144 TI - A biorobotic structural model of the mammalian muscle spindle primary afferent response. AB - A biorobotic model of the mammalian muscle spindle Ia response was implemented in precision hardware. We derived engineering specifications from displacement, receptor potential, and Ia data in the muscle spindle literature, allowing reproduction of muscle spindle behavior directly in the robot's hardware; a linear actuator replicated intrafusal contractile behavior, a cantilever-based transducer reproduced sensory membrane depolarization, and a voltage-controlled oscillator encoded strain into a frequency signal. Aspects of muscle spindle behavior not intrinsic to the physical design were added in control software using an adaptation of Schaafsma's mathematical model. We tuned the response to biological ramp and hold metrics including peak, mean, dynamic index, time domain response, and sensory region displacement. The model was validated against biological Ia response to ramp and hold, sinusoidal and fusimotor inputs. The response with dynamic or static gamma motorneuron input was excellent across all studies. The passive spindle response matched well in five of the nine measures. Potential applications include basic science muscle spindle research and applied research in prosthetics and robotics. PMID- 11874145 TI - Algorithm for the detection of muscle activation in surface electromyograms during periodic activity. AB - A simple surface electromyography (EMG) activation detection algorithm was developed for improved numerical definition of initiation and deactivation of muscle activity during periodic motion when maximum voluntary contractions are impractical to obtain. For the encapsulation of activation/ deactivation periods of a signal as percentages of normal cycle parameters, two interrelated and variable thresholds of percent amplitude and duration of a normalized cycle were the analyzed inputs into an algorithm. Outputs for statistical analysis were total percent activation per cycle, standard deviation of activity per cycle, and temporal indices of where the signal turned on and off. Percent activity per cycle had a coefficient of variance of 0.24 (0.11). After the user chose whether to consider the signal for either encompassing all non-base-line activity or peak activity only, resulting coefficients of variation for percent activity were reduced to 0.16 (0.08). The results indicated the feasibility of a mathematically simple algorithm for repeatable decomposition of EMG activity. The need for a modifiable threshold parameter to incorporate varying needs of salient activity levels was also substantiated. PMID- 11874146 TI - Non-invasive tests for acute pulmonary embolism: what are the real advances? PMID- 11874147 TI - Intermediate probability lung scans (IPLS): retrospective review of 82 cases. AB - OBJECTIVE: In the light of a reported 30-40% prevalence of pulmonary embolism (PE) in intermediate probability lung scans (IPLS) based on results of the Prospective Investigation of Pulmonary Embolism Diagnosis (PIOPED) study, we examined the frequency of documented PE in 82 patients with IPLS, the management strategy employed in these patients with regards to additional imaging (e.g. further evaluation with venous sonography or spiral computed tomographic angiography (CTA)), anticoagulation therapy, and subsequent follow-up outcomes. METHOD: Retrospective review of the medical records of 82 patients with intermediate probability ventilation-perfusion (V/Q) lung scans from January 1998 to July 1999. RESULTS: 14.1% of V/Q scans were reported as having an intermediate probability of PE. 72% of IPLS were subject to further evaluation with venous Doppler ultrasound and/or CTA, and 39% of these patients had evidence of thrombo embolic disease. All patients with imaging evidence of thromboembolic disease were started on anticoagulation therapy. In addition, 19 patients were treated based on clinical judgement. Amongst the 35 patients who were not treated, 17 (49%) were based on clinical findings without further imaging. There was no mortality on follow-up of 28 cases of untreated IPLS. CONCLUSION: The majority of IPLS will have further imaging, out of which over one-third will have thrombo embolic disease. Approximately half of IPLS cases will receive anticoagulation therapy. No mortality or PE was found on follow-up of patients who were not treated. PMID- 11874148 TI - The role of spiral computed tomogram in the diagnosis of acute pulmonary embolism. AB - Acute pulmonary embolism is associated with considerable morbidity and mortality. Early diagnosis and prompt treatment is essential. A number of non-invasive diagnostic tools are available for its detection. However, each one of these tests has its limitations and the invasive pulmonary angiography remains the gold standard. We describe the use of spiral volumetric computerised tomogram in the diagnosis of acute pulmonary embolism in six patients in our centre where ventilation-perfusion scan facility is not available. This safe, simple and non invasive test has an excellent sensitivity and specificity for the detection of central and segmental pulmonary embolism and may replace the conventional invasive pulmonary angiography for the diagnosis of pulmonary embolism. PMID- 11874149 TI - Percutaneous endoscopic gastrostomy--indications and outcome of our experience at the Singapore General Hospital. AB - INTRODUCTION: Percutaneous endoscopic gastrostomy (PEG) is widely used for patients with dysphagia from neurological causes and head and neck malignancy. We examined the indications, complication rates and long term outcome of PEG inserted in our department. METHODS: We performed a study of PEG inserted in our department between January 1995 to March 2000. Consecutive patients with PEG inserted during this period were identified from our database that contained demographic data, primary and secondary underlying medical conditions, and immediate complications after the procedure. Casenotes were reviewed and caregivers (relatives or staff at nursing homes) were contacted for information on long term outcome at the time of this study between April 2000. Data was collected in standard form designed for this study. RESULTS: 181 cases of PEG insertion were performed during the study period. 174 patients were successfully followed up and reviewed. The median age was 70.5 (range 24 to 93) years old and there were 111 males. Indications for PEG insertion were: cerebrovascular diseases (60.4%), Parkinson's disease and other neuromuscular disorders (10.9%), nasopharyngeal carcinoma and other upper gastrointestinal malignancies (24.7%), and head injury (4%). Superficial wound infection (22.4%) and granuloma formation (31%) were common minor complications. Major complications were infrequent: peritonitis (2.3%) and gastrointestinal bleeding (0.6%). The mortality rates were 11.5% and 28.2% at one and six months respectively. Only one death from peritonitis was directly attributed to the procedure, most deaths were due to underlying co-morbidities with pneumonia being the most common cause. The proportion of the first PEG tubes removed or replaced were 12.2% and 35.5% at one and six months respectively. Thirty tubes were replaced due to blockage at median interval of 9.6 months. 9.7% of PEG tubes functioned longer than 24 months. CONCLUSIONS: Our results confirm the safety of PEG tubes in elderly patients with multiple co-morbidities. Major complications of the procedure were infrequent but produced grave consequences in these elderly patients with multiple co morbidities. As such, patients considered for PEG feeding should have reasonable prognosis and the procedure is inappropriate for patients with rapidly progressive and incurable diseases. PMID- 11874150 TI - Falls amongst institutionalised psycho-geriatric patients. AB - Falls are common among the elderly patients in the psycho-geriatric wards and yet they have been understudied. A fall is a multi-factorial syndrome involving the patient and the environment. Psycho-geriatric patients who fall may suffer serious physical injuries that result in morbidity, further institutionalisation or even mortality. This study aims to examine the contributing factors to, and morbidity and outcome of falls among institutionalised psycho-geriatric patients so that preventive strategies can be refined. Data of patients who fell over a year's period in four psycho-geriatric wards were collected retrospectively and compared with those who had not fallen within the same period. The general profile of the psycho-geriatric patient who falls is one who is: above seventy five years old; on three or more medications; and having recent changes in medication and mental status. About one in three fallers fell repeatedly. The rate of serious injury and mortality was low. In conclusion, while many factors are attributable to the common effects of aging and physical illnesses; psychotropic medication, change in mental state and specific environmental factors also play significant contributory roles to falls in this group of patients. PMID- 11874151 TI - A ten-year review of ruptured sinus of valsalva: clinico-pathological and echo Doppler features. AB - Rupture of the sinus of valsalva (RSOV) is an uncommon condition with a variety of manifestations ranging from an asymptomatic murmur to cardiogenic shock. This retrospective 10-year review (1985-1995) of 18 patients from a single institution revealed that 6 (33%) were female and 12 (67%) were male with a mean age of 37.6 +/- 13.4 years and that 72% were Chinese by ethnic descent with the remaining 28% being Malay. Eight patients (44.4%) presented with an asymptomatic murmur, 4 (22.2%) with acute chest pain, 4 (22.2%) with mild heart failure, 2 (11.1%) with severe heart failure, and 2 (11.1%) with cardiogenic shock. Rupture of the right aneurysmal coronary cusp (RCC) made up 15 (83.3%) while those of the non-coronary cusp (NCC) made up the remaining. Most of the RCC ruptures were directed into the right ventricle and all of the NCC ruptures were into the right atrium. Ventricular septal defects (VSDs) were found in 9 (50%) of the patients, (although detected by echocardiography in only one third of those patients), aortic regurgitation in 6 (33.3%) and aortic valve vegetations in 2 (11.1%). Echocardiography was found to be accurate in diagnosing RSOVs with 100% diagnostic accuracy after 1990 with four misdiagnoses before 1990. Of these four patients, two were misdiagnosed as having VSDs, one as having a coronary arteriovenous fistula and one as having a patent ductus arteriosus. The anatomical structure of the "windsock" was seen in 64% of the patients who were correctly diagnosed. The pattern of colour flow and spectral Doppler was seen in all patients and helped to localise the site of rupture and the direction of flow. In summary, echocardiography is a simple and accurate way of diagnosing and defining RSOVs and is the imaging modality of choice. PMID- 11874152 TI - Cervical cord injury in an elderly man with a fused spine--a case report. AB - We report a case of an elderly man presenting with co-existing diffuse idiopathic skeletal hyperostosis (DISH) and ossified posterior longitudinal ligament (OPLL) resulting in central cord syndrome. Only three such cases have been reported co existing with DISH. The patient recovered most of his neurological deficit through conservative management. A discussion on the radiological features of DISH co-existing with OPLL and how these differ from ankylosing spondylitis (AS) follows. PMID- 11874153 TI - Adjuvant therapy of bullous pemphigoid with mycophenolate mofetil: old drug, new use. AB - Bullous Pemphigoid (BP) is an autoimmune subepidermal blistering disease appearing predominantly in the elderly. The disease is primarily treated with systemic corticosteroids. However, the treatment can be associated with significant morbidity. Adjuvant corticosteroid sparing therapy can also be associated with significant morbidity. In this study a case of BP which was difficult to control with systemic steroids was successfully treated with mycophenolate mofetil as adjuvant therapy. Mycophenolate mofetil used previously in transplantation, has recently been shown to be useful in autoimmune blistering disorders. Further study to confirm this significant finding and to determine if the long term prognosis of BP can be altered by the drug, is required. PMID- 11874154 TI - Herpes zoster ophthalmicus and the superior orbital fissure syndrome. AB - Herpes Zoster Ophthalmicus (HZO) is not an uncommon condition in the elderly and the immunocompromised. The common ocular manifestations include blepharoconjunctivitis, keratitis and uveitis. Dramatic presentations like orbital apex syndrome and superior orbital fissure syndromes occur rarely in patients with herpes zoster meningo-encephalitis. We report a patient with herpes zoster meningo-encephalitis and the superior orbital fissure syndrome (SOFS). PMID- 11874155 TI - Understanding traditional Chinese medicine--a doctor's viewpoint. AB - Singapore is a cosmopolitan country and its population comprises the Chinese, Malays, Indians, and others such as the Eurasians. In this heterogeneous, multi racial, multi-lingual and multi-cultural society, medical treatment is also varied. People can seek modern (mainstream, western) medicine or traditional medicine when they are sick. Usually they first seek modern medicine. Some turn to traditional medicine as complementary treatment or alternative treatment. Traditional medicine is here to stay in this country. In November 2000, the Traditional Chinese Medicine (TCM) Practitioners Bill was passed in the Singapore Parliament. Health care providers, including doctors, would benefit from a good knowledge of both modern and traditional medicine. Practitioners in traditional medicine should also learn modern medicine. PMID- 11874156 TI - Clinics in diagnostic imaging (66). Right complete branchial fistula. AB - We report a case of complete branchial fistula in an 18-year-old woman which could be demonstrated by a contrast study. The tract was completely excised and the patient had an uneventful recovery. Branchial fistulae are formed due to the persistence of the embryonic second branchial cleft. Complete fistulae with the internal opening in the region of the tonsillar fossa are not common. PMID- 11874157 TI - Legionella species infection in adult febrile respiratory tract infections in the community. AB - Legionella spp. (Lsp) are well recognized as etiologic factors in pneumonia but less so in respiratory tract infections (RTI) in the community. The objective of the present study was to characterize febrile RTI patients with a documented Legionella etiology, in terms of specific serogroups, clinical manifestations of the disease, disease course and the effect of antibiotic therapy. Ambulatory adults with febrile RTI (n = 250) were included in a prospective study in which the etiological causes of the infection were identified using sophisticated serological techniques. Paired sera were obtained for each of the patients and were tested for 41 different serotypes of Lsp using micro-immuno-fluorescence (MIF) serology. Only a significant change in IgG and/or IgM antibody titers was considered diagnostic. In 28 patients (11.2%) there was serological evidence of acute infection with 1 of the types of Lsp. The infections were manifested clinically as upper RTI in 9 patients and as lower RTI in the other 19 patients (community-acquired pneumonia in 2 of these). L. pneumophila serogroup 1 was identified in 3 patients, L. pneumophila serogroups higher than 1 were identified in 13 patients and L. non-pneumophila serogroups in 18 patients. The clinical and laboratory findings in patients with acute Lsp infection were not significantly different from those in patients without evidence of this infectious agent. The length and course of the disease were similar in the 12 patients treated with specific antibiotics for Lsp and in those who were not. We conclude that Lsp can be identified in a significant percentage of patients with acute febrile RTI. No specific clinical or laboratory features were observed for these patients and specific antibiotic therapy does not affect the course of the disease. PMID- 11874158 TI - Neisseria meningitidis with decreased susceptibility to penicillin in Istanbul, Turkey. AB - This study was conducted to estimate the rate of decreased susceptibility to penicillin (MIC > 0.06-1 microg/ml) in Neisseria meningitidis isolates in Istanbul, Turkey. A total of 30 isolates collected during a 1-y period from patients with meningitis and from nasopharyngeal carriers were tested for penicillin and cefotaxime susceptibility using the E-test. Two out of 12 (17%) clinical isolates and 11/18 (61%) nasopharyngeal isolates showed decreased susceptibility to penicillin with MICs in the range 0.094-1.0 microg/ml, giving an overall resistance of 43% (n = 13). These data show that continued surveillance of trends in antimicrobial susceptibility of N. meningitidis is important for detecting the emergence of N. meningitidis strains with MICs > 1 microg/ml which may pose serious therapeutic problems. PMID- 11874159 TI - Impact of combinations of antineoplastic drugs on intestinal microflora in 9 patients with leukaemia. AB - The impact of antineoplastic drugs on the intestinal microflora was studied in 9 patients with acute leukaemia during chemotherapy and in 5 patients also during chemotherapy-induced neutropenia. Quantitative and qualitative microbiological analyses of faecal samples obtained before and during chemotherapy showed significantly increased counts of Bacteroides spp. in 3/9 patients and, during neutropenia, significantly increased counts of yeasts in 2/5 patients; however, the intestinal microflora was stable in most patients. PMID- 11874160 TI - Chlamydia pneumoniae and severity of asthma. AB - A substantial increase in the prevalence of asthma in the Western world during the last few decades has led to a continuous search for novel factors that might be involved in the development of the disease. We carried out a study to clarify whether there is a relationship between severity of asthma and Chlamydia pneumoniae-specific titres at the group level and whether antibodies to the 60 kDa chlamydial heat shock protein (chsp60) are associated with asthma. A total of 116 (31 men, 85 women) consecutive asthma patients from a chest clinic were recruited and divided into 3 groups according to the severity of the disease: there were 13 asthmatics with severe, 54 with moderate and 49 with mild asthma. In addition, 50 (31 men, 19 women) consecutive blood donors were enrolled to serve as a control group. Sera for the measurements of specific IgG, IgA and IgM antibodies using a microimmunofluorescence test and of chsp60 using an enzyme immunoassay were obtained upon enrolment and also 3-4 months later from the asthma patients. Severe and moderate asthma were found to be strongly associated with elevated IgA antibody levels to C. pneumoniae [odds ratio (OR) 5.58, 95% confidence interval (CI) 1.31-23.72 for severe and OR 5.65, 95% CI 2.05-15.53 for moderate asthma] in a logistic regression model. Furthermore, in women, the occurrence of elevated IgA antibody levels and the age-adjusted geometric mean titres of IgA antibodies were significantly higher among the asthmatics than the controls (p = 0.003 and 0.04, respectively). Antibodies to chsp60 occurred more frequently and in higher concentrations among the asthmatics than the controls, although the differences did not reach significance. In conclusion, severe and moderate asthma were significantly associated with elevated IgA antibody levels to C. pneumoniae suggestive of chronic infection. Antibodies to chsp60 did not prove to be a useful marker of such an infection among the asthmatics studied here. PMID- 11874161 TI - Is the increase in notifications of Chlamydia trachomatis infections in Sweden the result of changes in prevalence, sampling frequency or diagnostic methods? AB - Based on clinical and laboratory surveillance data, trends in the incidence rates of genital Chlamydia trachomatis infections in Sweden between 1991 and 1999 were analysed according to age group and sex. The influence of changes in laboratory methods on the reported infections was assessed. After a decrease in the incidence rate of infection of 36% between 1991 and 1994, followed by a period of stability, a 20% increase was observed between 1997 and 1999 (from 157 to 189/100,000). Between 1991 and 1999 the female:male ratio decreased from 1.7 to 1.4. Incidence rates started to increase in 1994 in the 15-19 y age group for both sexes. Crude Chlamydia positivity increased from 4.1% (352,050 people tested) in 1994 to 5.4% (305,946 people tested) in 1999. This increase in Chlamydia positivity was seen both in laboratories that had changed to more sensitive methods and in those that had not. Changes in laboratory methods can therefore only partially explain the increase in notified cases. Increased screening of men may have contributed to the increase, but rising incidence rates in all young age groups of both sexes suggest a true increase in prevalence. PMID- 11874162 TI - Cervical ureaplasma urealyticum colonization: comparison of PCR and culture for its detection and association with preterm birth. AB - We developed a 16S ribosomal (r) RNA gene-based PCR assay specific for Ureaplasma urealyticum and compared it with culture. We also wanted to assess the role of cervical U. urealyticum colonization in preterm births. Cervical swabs from 100 women with preterm contractions and from 50 asymptomatic pregnant women were collected and analyzed using PCR. The PCR and culture methods were compared using the samples from the asymptomatic patients. PCR and culture were equally effective at detecting U. urealyticum. Cervical colonization correlated with preterm delivery, with rates of 71% and 37% for those delivering preterm and those with term delivery, respectively (p = 0.008). The relative risk of preterm delivery if colonized with U. urealyticum was 3.34. 16S rRNA gene-based PCR proved to be a useful tool for detecting U. urealyticum compared to culture. Lower genital tract colonization with U. urealyticum was associated with preterm birth. PMID- 11874164 TI - The naive CD4+ count in HIV-1-infected patients at time of initiation of highly active antiretroviral therapy is strongly associated with the level of immunological recovery. AB - Current antiretroviral therapy can induce considerable, sustained viral suppression followed by immunological recovery, in which naive CD4 + cells are important. Long-term immunological recovery was investigated during the first 3 y of highly active antiretroviral therapy (HAART) in 210 HIV-1-infected patients. The focus was on the naive CD4 + cell time course and associations between naive CD4 + cell counts and established prognostic markers. Total and naive CD4 + cell counts were measured using flow cytometry. The HIV-RNA detection limit was 20 copies/ml. During 36 months of HAART, the total CD4 + count followed a triphasic pattern, reflecting an initial phase of rapid redistribution from lymphoid tissues, followed by a slow increase, partially due to an increase in naive CD4+ cell count. From Month 18 onwards, both naive and total CD4 + cell counts stabilized, although viral suppression was sustained. There was no association between plasma viral load and the increase in naive CD4 + cell count. Importantly, baseline naive CD4 + cell count was significantly associated with the change in naive CD4 + cell count, suggesting that the naive cell count at baseline does influence the immunological recovery that can be obtained from treatment. Surprisingly, the naive CD4 + cell count tended to stabilize at a subnormal level after 18 months of HAART. This finding merits further investigation. PMID- 11874163 TI - Response to two consecutive protease inhibitor combination therapy regimens in a cohort of HIV-1-infected children. AB - The response to 2 consecutive protease inhibitor (P1) combination regimens was evaluated in a cohort of HIV-1-infected children. Twelve children, most of whom had been heavily treated, received a 3-drug treatment: saquinavir in hard gelatin capsules (SQVhgc) + zidovudine (ZDV) + didanosine. When this treatment failed it was replaced by a 4-drug regimen: ritonavir + SQVhgc + ZDV + lamivudine. A mild and temporary decrease in viral load (VL) was observed with the initial regimen (p = 0.22). Therapy failure occurred in 7 patients (58%) within 9 months and in another 3 (25%) within 9-18 months. The 7 children who failed within 9 months received the subsequent boosted regimen, leading to a significant and lasting reduction in VL (p = 0.001). None of the patients failed on the boosted regimen: 5/7 achieved a VL of < 400 copies/ml and 3/7 achieved a VL of < 50 copies/ml. Our results suggest that a 4-drug regimen including 2 PIs produces a better and more sustained response than a 3-drug regimen including only 1 PI, and that a good, sustained response is possible with subsequent boosted regimens even in heavily treated children. PMID- 11874165 TI - CMV disease in AIDS patients: incidence of CMV disease and relation to survival in a population-based study from Oslo. AB - CMV disease is an important cause of morbidity and mortality in patients with AIDS. The purpose of this study was to investigate the incidence of CMV disease in a well-defined population of AIDS patients with a high rate of autopsy. No such study has previously been published from Scandinavia. A total of 248 patients who developed clinical AIDS in Oslo during the period 1 January, 1983 to 31 December, 1995 were included. Autopsy was performed in 152 of 213 deaths (71.3%). CMV disease was diagnosed in 95 patients. In the autopsy group, 73 patients (48%) had CMV disease, and in 52 of these patients CMV disease was first detected at autopsy. Retinitis was the most frequent manifestation, followed by adrenalitis, pneumonitis, encephalitis and gastrointestinal disease. No intravenous drug users (IVDUs) were diagnosed alive with CMV disease. All patients diagnosed with CMV disease before death had evidence of CMV disease at autopsy despite anti-CMV treatment. CMV disease was associated with increased risk of death. We conclude that CMV disease was frequent in patients with AIDS during the study period, was associated with increased mortality and was often diagnosed too late for the administration of appropriate therapy. PMID- 11874166 TI - Appropriate antibiotic use according to diagnoses and bacteriological findings: report of 12 point-prevalence studies on antibiotic use in a university hospital. AB - The objective of this study was to evaluate antibiotic use in relation to diagnoses and bacteriological findings in a 600-bed Norwegian university hospital. Twelve point-prevalence studies of antibiotic use were conducted between 1996 and 1999. In the point-prevalence studies, 1,096 of 6,588 adult patients (16.6%) used on average 1.25 antibiotics each. Of the patients who received antibiotics, 35% were treated for hospital-acquired infections. Lower respiratory tract and urinary tract infections accounted for more than half of all antibiotic use. Pencillins represented 54% of antibiotic use, cephalosporins 9%, quinolones 6% and antifungal agents 0.7%. The prescribed daily doses for the penicillins were 2-3 times higher than the defined daily doses. Bacteriological samples were obtained from 929 (85%) patients. Compliance with the guidelines was > 90% and was highest when the results of bacteriological samples were positive. Good compliance led to low prevalence of antibiotic use and the use of narrow spectrum antibiotics. PMID- 11874167 TI - Streptococcus bovis meningitis in a healthy adult patient. AB - We describe the first case in the English language of Streptococcus bovis meningitis in a 45-y-old patient without any underlying disease or predisposing condition. S. bovis biotype II was isolated from his spinal fluid and blood. The illness was community-acquired and was clinically and biologically similar to disease caused by the classical meningeal pathogens. The patient was cured after 10 d of therapy with ceftriaxone and, 2.5 y later, is currently healthy. As a result of this case and a similar case published recently in the Spanish literature we conclude that S. bovis should be considered a microorganism capable of causing meningitis in the absence of any underlying condition or clear focus of infection. PMID- 11874168 TI - Streptococcus bovis meningitis in a neonate with Ivemark syndrome. AB - Although Streptococcus bovis infections in adults are associated with endocarditis and bowel neoplasms, S. bovis-associated meningitis is rare in neonates. We describe the case of a neonate with Ivemark syndrome, which possibly predisposed her to infection with this bacterium. PMID- 11874169 TI - Cephalic tetanus as a result of rooster pecking: an unusual case. AB - In this paper, a case of cephalic tetanus caused by rooster pecking to the face is presented. Cephalic tetanus is a rare type of tetanus defined by trismus and paralysis of 1 or more cranial nerves. On admission to hospital the patient had facial palsy and trismus. With proper medical management she recovered without any relapse. PMID- 11874170 TI - Anthrax meningitis: case report and review. AB - We report a case of meningitis caused by Bacillus anthracis. Although this agent appears to be a rare pathogen, it should always be considered in the differential diagnosis of haemorrhagic meningitis cases owing to the high mortality rate associated with it. PMID- 11874171 TI - Transient lupus anticoagulant and prolonged activated partial thromboplastin time secondary to Epstein-Barr virus infection. AB - We describe the case of a previously well 19-month-old boy who presented with an acute Epstein-Barr virus infection and a prolonged activated partial thromboplastin time (APTT) associated with the presence of a transient lupus anticoagulant (LA). The boy had an excellent outcome, with gradual normalization of the APTT and disappearance of the LA. PMID- 11874172 TI - Kikuchi Fujimoto disease: case report and review of the literature. AB - We report a case of Kikuchi Fujimoto disease in a 34-y-old woman, with emphasis on the clinical picture and pathologic findings. PMID- 11874173 TI - A case of persistent anemia in a renal transplant recipient: association with parvovirus B19 infection. AB - We report an unexplained anemia that persisted for 4 months in a renal transplant patient who was receiving immunosuppression therapy that included prednisolone, tacrolimus and azathioprine. A bone marrow biopsy demonstrated pure erythroid hypoplasia and occasional giant pronormoblasts with intranuclear inclusions, characteristic of a parvovirus B19 infection. Both the serum and bone marrow cells were positive by parvovirus B19 DNA PCR. The anemia resolved 6 weeks after the administration of intravenous immunoglobulin (IVIG). Four months later, anemia redeveloped and IVIG was infused again. Hemoglobin levels were, however, still subnormal after 1 month of treatment and tacrolimus was then switched to cyclosporin A, resulting in a clear improvement. A parvovirus B19 infection should be included in the differential diagnosis of renal transplant recipients who present with anemia associated with a low reticulocyte count. Tacrolimus may possibly impair the clearance of a parvovirus B19 infection. PMID- 11874174 TI - Two cases of tularaemia after an orienteering contest on the non-endemic Island of Bornholm. PMID- 11874175 TI - Deteriorating diabetes control during adolescence: physiological or psychosocial? AB - Metabolic control often worsens during the pubertal years in individuals with type diabetes mellitus (DM). This may be due in part to the developmental transition from dependency on parents to a more autonomous lifestyle. Adolescence is also marked by challenging of authority figures and previous ways of thinking, experimentation, and, in some female teenagers, emergence of disordered eating attitudes and behaviors. The insulin resistance of puberty may be particularly maladaptive for the teenager with type 1 DM and is likely under-recognized. In this review, we examine the impact of physiological and psychosocial changes during puberty in type 1 DM, highlight treatment strategies aimed at modifying these factors, and suggest further interventions that warrant evaluation. PMID- 11874176 TI - How should the short stature of girls with Turner's syndrome be treated? PMID- 11874177 TI - Favorable final height outcome in girls with Ullrich-Turner syndrome treated with low-dose growth hormone together with oxandrolone despite starting treatment after 10 years of age. AB - The objective of this study was to find out whether moderate doses of growth hormone (GH) in combination with oxandrolone (Ox) and late initiation of puberty could improve adult height even in relatively old patients with Ullrich-Turner syndrome (UTS). Ninety-one patients with UTS were randomly assigned to receive either GH alone (Saizen, Ares-Serono, Geneva) 18 IU/m2/week (0.2 mg/kg/week) by daily s.c. injections (group GH) or a combination of GH and Ox 0.1 mg/kg/day p.o. (group GH + Ox). Prior to treatment mean age was 10.2 years (GH) and 10.5 years (GH + Ox), mean projected adult height (PAH) was 146.4 cm (GH) and 146.7 cm (GH + Ox). During year 2 the GH dose was increased in the GH group to 24 and later to 28 IU/m2/week (0.27 mg and later 0.31 mg/kg/week). In group GH + Ox, the Ox dose was reduced to 0.05 mg/kg/day after the first 12 months of therapy, and during the last treatment years the GH dose was raised to 24-28 IU/m2/week (0.27-0.31 mg/kg/week) due to declining growth promotion. Some of the patients of group GH were later given Ox in addition to GH because of waning growth velocity, whereas some of the patients of group GH + Ox were taken off Ox due to virilizing side effects of the high Ox dose, thus making up a third group of patients: group GH + transient Ox. Puberty was induced at a mean age of 14.9 years. In group GH + Ox, cumulative growth during 5 years of therapy was twice the growth anticipated from standards of untreated patients with UTS. Forty-seven patients are now near or at final height: in group GH (n = 7), mean final height was 151.7 cm (PAH 148.1 cm, gain 3.6 cm); in group GH + Ox (n = 15), 155.1 cm (PAH 147.2 cm, gain 7.9 cm); and in group GH + transient Ox (n = 25), 152.8 cm (PAH 146.4 cm, gain 6.4 cm). These results should be regarded as an underestimate of true final height since some the patients are still growing. Moderate doses of GH plus Ox and late induction of puberty definitely improved final height even in patients with UTS treated relatively late. PMID- 11874178 TI - SHOX intragenic microsatellite analysis in patients with short stature. AB - BACKGROUND: SHOX haplo-insufficiency is considered the molecular basis of short stature in patients with Turner's syndrome, and gives rise to the short stature with mesomelic dysplasia and Madelung deformity of patients with Leri-Weill syndrome. OBJECTIVE: Analysis of the intragenic SHOX microsatellite to define its utility in detecting SHOX haplo-insufficiency in patients with short stature. PATIENTS AND METHODS: 207 patients with short stature (57 girls with Turner's syndrome [TS] [24 mosaicisms]; 73 children with isolated short stature [ISS]; 77 patients with short stature and skeletal disproportion) and 30 control subjects. DNA extraction and PCR amplification of the intragenic SHOX microsatellite, at the 5'-untranslated region. SSCP and partial sequencing of the SHOX gene in one patient with Madelung deformity and two SHOX alleles. DXS1055 (Xp) and DXS1192 (Xq) microsatellites were also analyzed, together with DXS233 and DXS234 at 0 and 2 cM of the pseudoautosomal region (PAR), in patients with one SHOX allele. RESULTS: 1. 93% of patients with TS had a single SHOX allele, and allele unbalance was detected in the remainder. 2. Patients with ISS were not different from the normal population with respect to SHOX heterozygosity (0.92 and 0.93, respectively; p = 0.997). 3. Patients with short stature and skeletal disproportion showed a higher frequency of SHOX homo/hemizygosity (0.27 vs 0.08; p = 0.027). 4. Five patients with short stature with SHOX haplo-insufficiency were detected: three had Madelung deformity (inherited Yq;Xp translocation, de novo PAR deletion, and SHOX microdeletion), and two had de novo/inherited Xp partial monosomy. CONCLUSIONS: The SHOX intragenic microsatellite might be a useful molecular marker to detect TS (including Xp distal deletions). SHOX haplo insufficiency seems not to be an important contributor to ISS, but when skeletal disproportion is associated with short stature, a significant proportion of patients is found to have a single SHOX allele. Some of these patients were found to be SHOX haplo-insufficient upon molecular, cytogenetic and radiological examination. PMID- 11874179 TI - Auxological computer based network for early detection of disorders of growth and weight attainment. AB - INTRODUCTION: For several years practising pediatricians and pediatric endocrinologists in Leipzig have been collaborating closely to achieve early detection of growth disorders. As a result of this collaboration a data-bank was established in September 1998 into which height and weight measurements were entered and related to age and gender. This well established network of pediatricians in practice, pediatric endocrinologists and the auxological data bank functions well. METHODS: All data are anonymized and continually monitored. By September 2000, the data for 60,984 children had been assessed. The data were evaluated using the German Synthethic Norm Curve for body height and German normal data for BMI. Pathology of growth dynamics was assumed when change in height SDS was greater than 0.5 height SDS/year. RESULTS: Analysis showed that 2,216 children (3.6%) had a height greater than the 97th percentile and 2,775 children (4.5%) had a height less than the 3rd percentile of the normative reference. BMI above the 97th percentile was found in 7,687 children (12.6%) and below the 3rd percentile in 2,678 children (4.4%). When assessing growth development (n = 5,665), 194 (3.4%) showed acceleration and 155 (2.7%) showed deceleration of growth. CONCLUSION: The computer-based monitoring system is useful to document short-term and long-term changes of growth and weight attainment in the general population. The presence of growth disorders and/or disturbed weight gain can be detected early in the individual child. PMID- 11874180 TI - Microadenomas of the pituitary gland in children with and without hypophyseal dysfunction in magnetic resonance imaging. AB - The aim of this study was to correlate lesions of the pituitary gland with hormonal dysregulation. The hormonal status of 63 children was correlated with MRI findings of the pituitary gland. Two radiologists judged the MRI examinations without knowledge of the hormonal situation. The reliability of the diagnosis "adenoma" was evaluated in five steps from 0-100% for each sequence. A microadenoma was found in six of 14 children with hyperprolactinemia and in six of eight patients with increased IGF-I/IGFBP-3. However, microadenomas were also detected in eight of 28 children without hormonal dysfunction (clinical feature: obesity). The adenomas were seen best in a dynamic sequence after gadolinium administration. An expansive growing macroadenoma was found in one of 13 patients with hypopituitarism. We found a relatively high number of microadenomas even in children without any hormonal dysfunction. Taking into account the reported autopsy results (6.1-27% occult microadenomas), we suggest that the MRI diagnosis "microadenoma" is made too frequently if usual MRI criteria are used. Patients with increased levels of IGF-I/IGFBP-3 had a high incidence of microadenoma (up to 87.5%). Hyperprolactinemia was associated with microadenomas in about 43% ( 57%) of patients (nearly on the same level as children without hormonal dysfunction). Therefore unspecific stimulation of the pituitary gland with consecutive increased volume seems to be responsible for hyperprolactinemia in many of these patients. PMID- 11874182 TI - The relationship between clinical severity of Noonan's syndrome and growth, growth hormone (GH) secretion and response to GH treatment. AB - Short stature is one of the major features of Noonan's syndrome (NS). In a multicentre trial of growth hormone (GH) therapy in 25 children with NS, we observed a large inter-individual variation in first-year response to GH treatment. This suggested that subgroups might exist in NS that differ in either endogenous GH status or responsiveness to GH therapy. We therefore related growth, GH secretion and 2 years response to GH treatment to subtypes of phenotypic expression of NS. Twelve patients were moderately affected and 13 had a severe clinical phenotype of NS. The variability in phenotype did not correlate with significant differences in intra-uterine growth, infancy growth or childhood growth, and response to GH treatment. However, the variability in phenotype severity did account for striking differences in endogenous GH secretion. PMID- 11874181 TI - Pamidronate treatment of osteogenesis imperfecta--lack of correlation between clinical severity, age at onset of treatment, predicted collagen mutation and treatment response. AB - Severe forms of osteogenesis imperfecta (OI) are characterised by osteoporosis with multiple fractures, deformity, progressive loss of mobility and chronic bone pain. Bisphosphonates, as osteoclast inhibitors, reduce bone turnover and improve osteoporosis. OBJECTIVE: To investigate the effect of pamidronate treatment of severe OI in children, and find any correlation between clinical severity, age at start of treatment, type of predicted collagen mutation and treatment response. DESIGN: Open, observational trial. PATIENTS: A two-year study of pamidronate treatment was undertaken in a cohort of 18 children, (1.4-14.5 years) with OI types III and IV. INTERVENTIONS: Disodium pamidronate, 1 mg/kg/day for 3 days every 4 months, by i.v. infusion with measurement of bone turnover, bone density, vertebral morphology and skin biopsies to assess collagen mutation. RESULTS: Eleven children have completed 2 years of treatment and three more have completed 20 months. Sustained cessation of bone pain, improved mobility and decreased fracture rate were seen in all patients. Bone turnover decreased slightly but was not statistically significant. Bone mineral density (BMD) of lumbar spine increased by a mean of 124.7 +/- 75.7% over 2 years (Z score mean -5.08 +/- 1.27, to -3.30 +/- 1.71, p <0.001); the greatest change in BMD was seen in the most severely affected patients: 138 +/- 50.6% (severe), 62.47 +/- 22.9% (mild). There was a mean increase in vertebral height at L4 of 68.5% and in vertebral area of 85.4%. The majority of patients had slow electrophoretic migration of type I collagen alpha chains or reduced secretion of type I collagen, indicative of structural, helix-breaking mutations. There was no correlation between phenotypic severity, age at start of treatment and treatment response (r2 = 0.14) CONCLUSIONS: Pamidronate treatment of severe forms of OI is an effective therapeutic modality to increase bone density, decrease fracture rate, increase mobility and improve quality of life, irrespective of the severity of the mutation or clinical phenotype. It has a good short-term safety profile. PMID- 11874183 TI - Cardiac mass and function, carotid artery intima-media thickness and lipoprotein (a) levels in children and adolescents with type 1 diabetes mellitus of short duration. AB - OBJECTIVE: To evaluate cardiac mass and function, carotid intima-media thickness, and serum lipid and lipoprotein (a) (Lpa) levels in children and adolescents with type 1 diabetes mellitus (DM) of short duration. BACKGROUND: Diabetes mellitus has been found to be an important risk factor for macrovascular disease in adults. Increased serum lipids and Lpa levels have been reported in adolescents with type 1 DM; atherosclerotic vascular lesions involving a combination of fatty degeneration and vessel stiffening of the arterial wall and myocardial involvement impairing diastolic function may be present in adolescents and young adults with type 1 DM. DESIGN/METHODS: Twenty children and adolescents (10 males, 10 females) diagnosed with type 1 DM before 3.4 +/- 3.3 years with a mean age of 11.9 +/- 3.6 years were studied; their HbA1c levels were 8.0 +/- 1.9%. Twenty healthy non-diabetic controls, 10 males and 10 females, aged 12.1 +/- 3.4 years, matched for height and weight, participated in the study. Fasting blood samples were obtained for lipid and Lpa analysis. Patients underwent transthoracic M-mode and two-dimensional echocardiographic evaluation for measurement of left atrial and ventricular dimensions and left ventricular (LV) wall thickness and mass. Stroke volume and cardiac output were measured using pulsed Doppler echocardiography; carotid intima-media thickness was measured using high resolution mode B ultrasound. RESULTS: Interventricular septal thickness (7.1 +/- 1.8 vs 7.0 +/- 1.5 mm), LV posterior wall thickness (7.1 +/- 1.4 vs 7.5 +/- 2.0 mm) and LV mass after correction for body surface area (70.6 +/- 27.4 vs 70.7 +/- 18.0 g/m2) were similar in patients and controls. Similarly, the LV ejection fraction at rest was similar in patients and controls (69.9 +/- 2.3 vs 70.0 +/- 0.6%), as were pulmonary venous flow velocities (0.56 +/- 0.09 vs 0.55 +/- 0.10 m/s for diastolic peak velocity, 0.54 +/- 0.08 vs 0.50 +/- 0.09 m/s for systolic peak velocity and 0.17 +/- 0.07 vs 0.19 +/- 0.05 m/s for atrial reversal filling). Carotid intima-media thickness (0.60 +/- 0.02 and 0.59 +/- 0.02 mm for the right and left carotid artery) was similar to that of controls (0.60 +/- 0.03 and 0.61 +/- 0.02 mm for the right and left carotid artery). Low density lipoprotein cholesterol and Lpa levels were increased in patients compared to controls (113.2 +/- 26.0 mg/dl and 20.1 +/- 11.7 mg/dl in patients vs 90.4 +/- 14.3 mg/dl and 9.8 +/- 2.9 mg/dl in controls; p <0.01), while total cholesterol, HDL cholesterol and serum triglyceride concentrations were similar to those in controls. CONCLUSIONS: Although children and adolescents with type 1 DM seem not to show alterations in cardiac mass and function or early atherosclerotic changes in the first few years after diagnosis, their cardiovascular risk is increased as they present with dyslipidemia at an early stage of the disease. PMID- 11874184 TI - Tumor necrosis factor-alpha levels in short children. AB - Growth hormone has been suggested to modulate the release of cytokines, such as tumor necrosis factor-alpha (TNFalpha) and interleukin-1 (IL-1). Moreover, TNFalpha synthesis has been shown to be decreased in hypophysectomized rodents. The aim of this study was to evaluate the influence of GH status on TNFalpha levels in a group of 44 short prepubertal children. Among them, 13 children aged 9.8 +/- 3.5 years were growth hormone (GH) deficient and the other 31 short children had normal growth velocity, normal GH response to provocative testing, and did not suffer from any chronic disease, thus this group was diagnosed as having idiopathic short stature (ISS). A group of 40 age- and sex-matched healthy children was used as controls. No significant differences in basal TNFalpha levels (pg/ml) were found between the GH deficient, ISS children and healthy controls. Furthermore, there was no correlation between TNFalpha and basal serum concentrations of GH or peak GH levels after stimulation. Similarly, TNFalpha values did not correlate with either IGF-I or IGFBP-3 serum concentrations. PMID- 11874185 TI - Reference values for morning salivary cortisol concentrations in healthy school aged children. AB - INTRODUCTION: In children, sensitive, specific, pain-free sampling methods are important. An alternative is salivary sampling. Our knowledge about steroid levels in saliva and plasma in school-aged children and during puberty is sparse and contradictory. AIM OF THE STUDY: To estimate salivary cortisol concentrations in healthy school-aged children and relate the concentrations to age, sex, stage of puberty and adult values. PATIENTS AND METHOD: Saliva was collected in Salivette tubes from 210 boys and 176 girls aged 7-15 years, and from four adults, between 08.00 and 09.00 h. The tubes were centrifuged and then frozen at 20 degrees C until analysed by a commercial RIA-cortisol kit. RESULTS: Salivary cortisol was measurable in all infants. Boys and girls had nearly the same median concentrations, 8.8 versus 8.6 nmol/l, but girls had a higher maximal level, 53.9 compared to 33.2 nmol/l in boys. The median concentration was lower in 7-9 year old children, 7.2 in boys and 5.7 nmol/l in girls, compared to 10-12 year-old children, 11.5 in boys and 10.9 nmol/l in girls (p <0.001). The median concentrations in most age groups were lower than in adults. Salivary cortisol concentration was dependent on stage of puberty. CONCLUSION: Salivary cortisol was measurable in the morning in school-aged children and the median concentration was dependent on age, stage of puberty, but not on sex. PMID- 11874186 TI - Impaired glucose homeostasis in young adult thalassemic patients: a pilot study with acarbose. AB - This study investigated the effects of the alpha-glucosidase inhibitor, acarbose, on glycemic control and insulin secretion in thalassemic patients with impaired glucose tolerance. The safety and tolerability of the drug were also evaluated. Nine patients (4 men and 5 women, aged 20-34 years) with beta-thalassemia major received a standardized nutritional test load prior to and following 3 months treatment with acarbose 100 mg t.i.d. Blood glucose, insulin and C-peptide levels were measured at 0, 60, 90 and 120 min post-loading. Plasma glucose levels after 3 months of acarbose treatment tended to be slightly lower than pre-treatment levels. Although fasting serum insulin and plasma C-peptide levels were unchanged after acarbose therapy, postprandial serum levels of both hormones were markedly reduced (by 24-47% and 19-32%, respectively, at 60-120 min post-loading). Body mass index, liver enzymes and serum lipids were unaltered following acarbose treatment. Gastrointestinal disturbances were mild and tended to decrease during the course of acarbose therapy. Acarbose is a well-tolerated agent for the management of thalassemic patients with glucose intolerance and normal or increased insulin secretion. It is possible that acarbose may prevent or delay progression from impaired glucose homeostasis to frank insulin-dependent diabetes mellitus. PMID- 11874187 TI - An activating mutation of the thyrotropin receptor gene in hereditary non autoimmune hyperthyroidism. AB - The thyroid stimulating hormone (TSH) receptor gene displays a diverse spectrum of activating and inactivating mutations. We report a germline activating mutation M463V of the TSH receptor gene in two siblings with hereditary non autoimmune hyperthyroidism. The onset of disease in the affected members of the pedigree occurred during childhood or adolescence. The significance of diagnosing activating TSHR mutations lies in therapeutic management and genetic counseling; thyroid ablation is advocated as first line treatment. PMID- 11874188 TI - Growth hormone therapy may increase fracture risk in a pubertal patient with osteogenesis imperfecta. AB - We describe a patient with osteogenesis imperfecta type 1A who was treated with growth hormone for 9 years. The treatment resulted in a marked improvement in growth velocity, but four fractures occurred during the pubertal period. PMID- 11874189 TI - Hypogonadotrophic hypogonadism associated with prelingual deafness due to a connexin 26 gene mutation. AB - In Mediterranean countries, almost half the incidence of non-syndromic congenital hearing loss is caused by mutations in the gap junction (GJ) connexin 26 gene (GJB2/DFNB1 locus). In this form of deafness the cochlear defect is usually isolated. We describe here the first case of hypogonadotrophic hypogonadism in association with this particular cochlear defect. The male patient had moderate deafness inherited from his deaf parents. All family members had a homozygous 35delG mutation in the connexin 26 gene. This mutation accounts for 70% of all connexin 26 gene mutations. The patient was referred to a paediatric endocrinology unit at 11 years of age for moderate growth retardation. Growth rate was normal until 11 years. The patient then presented delayed puberty (testicular volume 4 ml, penis length 4 cm) and did not undergo the usual pubertal growth spurt. LH and FSH secretory responses to GnRH at the age of 14.5 years (bone age 13.5 years), were: LH baseline level 1.1 IU/l, peak 34 IU/l; FSH baseline level 1.8 IU/l, peak 5.7 IU/l. Testosterone concentration was <0.11 ng/ml. From 11 to 14 years old, testosterone concentration ranged from 0.11 to 0.2 ng/ml. Anti-Mullerian hormone (AMH) level was 38.6 ng/ml (normal for Tanner stage I), cortisol 109 ng/ml, and ACTH 37 pg/ml., Karyotype was 46 XY. On MRI analysis, the anterior pituitary and olfactory bulbs were normal. These data were consistent with partial hypogonadotrophic hypogonadism of hypothalamic origin, and the patient was treated with testosterone. This report supports the possible involvement of connexins in puberty initiation. Connexins may play a part in the co-ordination and synchronisation of GnRH release. PMID- 11874190 TI - Relation between parental restrictions on movies and adolescent use of tobacco and alcohol. AB - CONTEXT: Viewing smoking and drinking in movies may prompt adolescents to initiate these behaviors. Movies with R ratings contain more smoking than do movies in all other rating categories. OBJECTIVE: To evaluate the extent to which parents restrict the exposure of adolescents to R-rated movies and to determine whether such restrictions are associated with decreased tobacco and alcohol use in adolescents. DESIGN: Cross-sectional, self-administered survey. PARTICIPANTS: Students in grades 5 through 8 enrolled in New Hampshire and Vermont schools. SAMPLING STRATEGY: Fifteen schools in Vermont and New Hampshire were randomly selected from all middle schools with > or = 150 students. OUTCOME MEASURES: Students who had tried smoking cigarettes or tried drinking alcohol without parental knowledge. RESULTS: Of 4544 students surveyed, 18% had tried cigarettes and 23% had tried alcohol. Although 90% were younger than 14 years of age, only 16% were completely restricted from viewing R-rated movies. The prevalence of having tried smoking was 35% for those with no restrictions on viewing R-rated movies, 12% for those with partial restrictions, and 2% for those with complete restrictions. The prevalence of having tried alcohol was 46% for those with no restrictions on viewing R-rated movies, 16% for those with partial restrictions, and 4% for those with complete restrictions. Even after controlling for other factors, including grade, parental disapproval of smoking, maternal supervision, maternal responsiveness, peer and family smoking, and child personality characteristics, children who were completely restricted from viewing R-rated movies were significantly less likely to smoke (relative risk, 0.29; 95% CI, 0.19 to 0.45) or drink (relative risk, 0.30; CI, 0.21 to 0.42) compared with those who had no restrictions on viewing R-rated movies. CONCLUSION: Limiting the exposure of adolescents to R-rated movies may prevent early use of alcohol and tobacco. PMID- 11874191 TI - Outpatient diagnostic errors: unrecognized hyperglycemia. AB - CONTEXT: To estimate the prevalence of unrecognized diabetes in a large managed care organization (MCO). DESIGN: Retrospective data analysis. PATIENTS AND SETTING: All patients over age 30 enrolled in the (staff-model) MCO of Duke University Medical Center between April 1996 and March 1999. DATA SOURCES: Merged database of MCO administrative, billing, and laboratory files and selected medical records. CASE DEFINITIONS: We identified all patients with abnormal test results suggestive of diabetes (i.e., hemoglobin [Hb] A1c > or = 7.0% or plasma glucose > or = 200 mg/dL) on one or more occasions. Patients were considered to have recognized diabetes if they had an ICD-9 diagnostic code for diabetes in the administrative database or a diagnosis mentioned in their medical record (on the basis of medical record review of a random sample of 30% of patients with abnormal test results and no ICD-9 code). Patients with unrecognized diabetes did not have an ICD-9 code or a medical record diagnosis. RESULTS: 1426 patients had laboratory tests suggestive of diabetes. Of these patients, 1122 (79%) had an ICD 9 diagnostic code for diabetes in the administrative database. Forty-six of the remaining 304 patients without ICD-9 codes had mention of diabetes on medical record review; thus, we estimate that as many as 258 (18% of patients with laboratory tests suggestive of diabetes) had unrecognized diabetes. When this estimate was restricted to findings that are most suggestive of diabetes (high HbA1c or two high plasma glucose tests), 124 (9%) patients had unrecognized diabetes. CONCLUSIONS: A substantial proportion of patients in an MCO have laboratory values suggestive of diabetes with no evidence that their providers have recognized this condition. PMID- 11874192 TI - Determining medical error. Three case reports. PMID- 11874193 TI - Smoke-free movies: sense or censorship? PMID- 11874194 TI - Rate movies with smoking "R". PMID- 11874195 TI - Numeracy and the medical student's ability to interpret data. AB - CONTEXT: Although the ability to work with numbers is important to the practice of medicine, little is known about physician numeracy (basic skill with numbers). OBJECTIVE: To test medical students' numeracy and how it relates to the ability to interpret risk-reduction information. DESIGN: Randomized, cross-sectional survey. SAMPLE: 62 first-year medical students at the University of North Carolina at Chapel Hill medical school who attended a risk-communication seminar and had usable survey data (46% of the 134 students who received the survey). INTERVENTION: Students were given information about the baseline risk for developing a hypothetical disease and were randomly assigned to one of four risk reduction presentations-relative risk reduction, absolute risk reduction, number needed to treat (NNT), or a combination of these three formats-about how two drugs would reduce this risk. OUTCOME MEASURES: Number of correct answers to three numeracy questions (stating that 500 heads would be expected in 1000 coin flips; converting "1% of 1000" to 10; and converting "1 in 1,000" to 0.1%). Correct data interpretation was judged with two tasks: a comparative task (i.e., state which drug provides greater benefit) and a quantitative task (i.e., calculate how much one of the drugs reduces disease risk). RESULTS: 77% of students answered all three numeracy questions correctly; 18% answered two correctly; and 5% answered one or none correctly. While 90% correctly stated which drug worked better, only 61% accurately interpreted the quantitative data. The ability to interpret data varied with numeracy: 71% of students who answered all three numeracy questions correctly also accurately interpreted the quantitative data, compared with 36% who answered two questions correctly and 0% who answered one or no questions correctly (P < 0.01). Correct quantitative interpretation was lower with the NNT format than with the other three formats (25% vs. 75%; P = 0.01). CONCLUSIONS: Almost one quarter of first-year medical students in our study had trouble performing basic numerical tasks. Those who had trouble also seemed to have difficulty interpreting medical data. This difficulty seemed to be exacerbated by presenting data in the NNT format. PMID- 11874196 TI - Primer on interpreting surveys. PMID- 11874197 TI - Promoting self-referral for laboratory testing. PMID- 11874198 TI - Glutamate receptor desensitization block potentiates the stimulated GABA release through external Ca2+-independent mechanisms from granule cells of olfactory bulb. AB - Glutamate stimulated release of [3H]GABA was studied, during receptor desensitization block and its modulation by voltage gated Ca2+ channels, internal Ca2+ mobilization and GABA transport inhibitors from olfactory bulb slices. Under control conditions, glutamate and agonists induced release was strongly inhibited by Mg/0 Ca2+ Krebs and Cd2+ and partially inhibited by Ni2+ and nifedipine. Cyclothiazide, which blocks desensitization of glutamate receptors, potentiated glutamate, kainate, AMPA and quisqualate induced release. This effect was less dependent of entry of external Ca2+, but was inhibited by trifluoperazine and thapsigargin, inhibitors of Ca2+-calmodulin and endoplasmatic Ca2+ ATPase respectively. Nipecotic acid and NO-711, inhibitors of the GABA transporter, were also able to reduce cyclothiazide potentiated release induced by the 4 secretagogues. Under control conditions, glutamate stimulates the release of GABA in cooperation with VDCC. However, during receptor desensitization block, glutamate stimulated GABA release is mainly modulated through mechanisms dependent on internal Ca2+ mobilization and reversal of the GABA transporter. PMID- 11874199 TI - Role of reactive oxygen species and glutathione in inorganic mercury-induced injury in human glioma cells. AB - The present study was undertaken to examine the role of reactive oxygen species (ROS) and glutathione (GSH) in glia cells using human glioma cell line A172 cells. HgCl2 caused the loss of cell viability in a dose-dependent manner. HgCl2 induced loss of cell viability was not affected by H2O2 scavengers catalase and pyruvate, a superoxide scavenger superoxide dismutase, a peroxynitrite scavenger uric acid, and an inhibitor of nitric oxide N(G)-nitro-arginine Methyl ester. HgCl2 did not cause changes in DCF fluorescence, an H2O2-sensitive fluorescent dye. The loss of cell viability was significantly prevented by the hydroxyl radical scavengers dimethylthiourea and thiourea, but it was not affected by antioxidants DPPD and Trlox. HgCl2-induced loss of cell viability was accompanied by a significant reduction in GSH content. The GSH depletion was almost completely prevented by thiols dithiothreitol and GSH, whereas the loss of viability was partially prevented by these agents. Incubation of cells with 0.2 mM buthionine sulfoximine for 24 hr, a selective inhibitor of gamma glutamylcysteine synthetase, resulted in 56% reduction in GSH content without any change in cell viability. HgCl2 resulted in 34% reduction in GSH content, which was accompanied by 59% loss of cell viability. These results suggest that HgCl2 induced cell death is not associated with generation of H2O2 and ROS-induced lipid peroxidation. In addition, these data suggest that the depletion of endogenous GSH itself may not play a critical role in the HgCl2-induced cytotoxicity in human glioma cells. PMID- 11874200 TI - Inhibition of rat brain Na+, K+-ATPase activity induced by homocysteine is probably mediated by oxidative stress. AB - The objective of the present study was to investigate the effects of preincubation of hippocampus homogenates in the presence of homocysteine or methionine on Na+, K+-ATPase and Mg2+-ATPase activities in synaptic membranes of rats. Homocysteine significantly inhibited Na+, K+-ATPase activity, whereas methionine had no effect. Mg2+-ATPase activity was not altered by the metabolites. We also evaluated the effect of incubating glutathione, cysteine, dithiothreitol, trolox, superoxide dismutase and GM1 ganglioside alone or incubation with homocysteine on Na+, K+-ATPase activity. Tested compounds did not alter Na+, K+-ATPase and Mg2+-ATPase activities, but except for trolox, prevented the inhibitory effect of homocysteine on Na+, K+-ATPase activity. These results suggest that inhibition of this enzyme activity by homocysteine is possibly mediated by free radicals and may contribute to the neurological dysfunction found in homocystinuric patients. PMID- 11874201 TI - In vivo gene electroporation of glial cell line-derived neurotrophic factor (GDNF) into skeletal muscle of SOD1 mutant mice. AB - Motor neurons degenerate with intracellular vacuolar change and eventually disappear in spinal cords of SOD1 mutant mice, resembling human amyotrophic lateral sclerosis (ALS). The GDNF gene was electroporatically transferred into the leg muscles of SOD1 mutant mice and expressed in muscle cells. This gene therapy with GDNF delayed the deterioration of motor performance, being retrogradely transported into spinal motor neurons. However, the number of the motor neurons and survival of the mutant mice were not improved by GDNF treatment. These results indicate that in vivo gene electroporation of GDNF into muscles could be an appropriate therapeutic approach to ameliorate an early dysfunction of motor neurons in SOD1 mutant mice, but further improvement is needed to use this gene transfer as an effective treatment of ALS. PMID- 11874202 TI - Effects of microenvironment on morphology and function of the microglial cell line BV-2. AB - Effects of microenvironmental changes were examined in the microglial cell line BV-2. In serum supplemented medium cells were ameboid shaped and exhibited thin cytoplasmatic processes at lower concentration or in absence of serum. High levels of acetylated low-density lipoprotein (LDL) receptor and of phagocytic and proliferative activity were detected. Lipopolysaccharide (LPS) and the neuropeptide substance P (SP) induced secretion of interleukin-6. Low interleukin 3 secretion was detected only occasionally and was not influenced by LPS and SP. In defined medium, "process-bearing" cells were evident. Compared to cultures in serum supplemented medium, the cells expressed lower acetylated LDL-binding and phagocytic activity while actively proliferated, the response to LPS was reduced and to SP absent. Granulocyte/macrophage colony-stimulating factor increased the number of process-bearing cells, of acetylated LDL-binding and of IL-6 secretion induced by LPS. Cell morphology was not influenced by neurotrophins like nerve growth factor and brain-derived neurotrophic factor. The described phenotypical and functional plasticity makes the BV-2 cell line a useful model to investigate mechanisms of microglial activation. PMID- 11874204 TI - Immunohistochemical study of immunophilin 1-15 fragment in intact frog brain, and in the brain and spinal cord of intact and spinal cord hemisectioned rats. AB - Previously by immunohistochemical technique the distribution of immunophilin 1-15 fragment (IphF) isolated from bovine hypothalamus was examined in various tissues (heart, lung), including immune system organs (spleen and thymus) of intact rats. IphF-like immunoreactivity (IphF-LI) was revealed in several cell types: lymphocytes, monocytes, macrophages and mast cells. In the present study the immunohistochemical localization of IphF was examined in intact rat and frog brains. In rat brain several cell groups concentrated particularly in the supraoptic nucleus (SON) of hypothalamus, medulla oblongata (reticular formation, olives, hypoglossal and facial motor nuclei) and cerebellum (lateral cerebellar nucleus) demonstrated IphF-LI. In frog hypothalamus (SON) the same working dilution (1:5000) of IphF-antiserum revealed very strong immunoreactivity. In the paraventricular nucleus (PVN) IphF-LI varicosities were scattered around the immunonegative cells. The second cell groups showing IphF-LI in the frog brain were gliocytes (mainly the astrocytes). Besides, IphF distribution was investigated in rats subjected to hemisection of spinal cord (SC) with and without administration of proline-rich polypeptide (PRP). PRP was isolated from bovine neurohypophysis neurosecretory granules, produced by magnocellular nuclei of hypothalamus. Hemisection of SC led to changes of IphF distribution in the hypothalamus. In PRP treated animals IphF showed no immunoreactivity. PRP is suggested to act as a neurotransmitter and neuroregulator. PMID- 11874203 TI - Effects of metabotropic glutamate receptor agonists and antagonists on D aspartate release from mouse cerebral cortical and striatal slices. AB - The cytosolic release of L-glutamate has been held to be responsible for the increase in extracellular glutamate to toxic levels in the brain. The mechanism and regulation of this release was now studied in cerebral cortical and striatal slices with D-[3H]aspartate, a non-metabolized analogue of L-glutamate and a poor substrate for vesicular uptake. L-Glutamate and D-aspartate strongly stimulated the release in a concentration-dependent manner. Of the ionotropic glutamate receptor agonists, only kainate enhanced the basal release in the striatum. Of the metabotropic glutamate receptor ligands, the group I agonist (S)-3,5 dihydroxyphenylglycine (S-DHPG) failed to affect the basal release but inhibited the D-aspartate-evoked release in the striatum. The group I antagonist (RS)-1 aminoindan-1,5-dicarboxylic acid (AIDA) had no effect on the basal release in either preparation but enhanced the L-glutamate-evoked release and inhibited the D-aspartate-evoked release in the striatum, not however in the cerebral cortex. The group II agonist (2S,2'R,3'R)-2-(2',3'-dicarboxycyclopropyl)glycine (DCG IV) and the group II antagonist (2S)-2-ethylglutamate (EGLU) were without effect on the basal, D-aspartate- and L-glutamate-evoked releases of D-[3H]aspartate in either preparation. The group III agonist L-serine-O-phosphate (L-SOP) failed to affect the basal release but reduced the D-aspartate-evoked release in the striatum. The group III antagonist (RS)alpha-methylserine-O-phosphate (MSOP) failed to affect the basal release but increased the glutamate-evoked release and inhibited the D-aspartate-evoked release in the striatum. Both L-trans pyrrolidine-2,4-dicarboxylate (L-trans-PDC) and (2S,1'S,2'R)-2 carboxycyclopropyl)glycine (L-CCG-III), transportable inhibitors of the high affinity glutamate uptake, enhanced the basal release, more strongly in the striatum than in the cerebral cortex. L-CCG-III also increased the L-glutamate evoked release in the striatum. Nontransportable dihydrokainate enhanced the basal release much less and failed to affect the glutamate-evoked release. The results indicate that the release of glutamate from cytosolic pools is carrier mediated via homoexchange. This process is regulated in the striatum by metabotropic group I and group III receptors in a manner different from the regulation of the vesicular release of glutamate from presynaptic terminals. PMID- 11874205 TI - Regulation of sulfoglucuronyl glycolipid synthesis in the developing rat sciatic nerve. AB - Sulfoglucuronyl glycolipids (SGGLs) have been considered as target antigens in demyelinating peripheral neuropathies associated with IgM monoclonal gammopathy. The regulation of expression of SGGLs in the rat sciatic nerve during development was studied by assaying the levels of SGGLs and activities of four glycosyltransferases sequentially involved in their synthesis from lactosylceramide. The levels of SGGLs in the sciatic nerve increased with development and reached a maximum at sixty days after birth. The rate of increase in the level of SGGLs between day 5 to 20 was similar to rate of deposition of myelin in the nerve. Analysis of the activities of the glycosyltransferases showed that only lactotriosylceramide galactosyltransferase (LcOse3Cer-GalTr) increased in parallel with the levels of SGGLs during development. The other three enzymes were not co-relative with the synthesis of SGGLs. The product of LcOse3Cer-GalTr reaction, nLcOse4Cer is the key intermediate for all neolactoglycolipids, particularly NeuAc alpha2-3nLcOse4Cer or nLM1, which is the major ganglioside (60%) of myelin in rat sciatic nerve. The results suggest that in the sciatic nerve SGGLs are mostly associated with Schwann cell myelin and their synthesis is regulated by LcOse3Cer-GalTr, unlike in the cerebral cortex and cerebellum where SGGLs are associated with the neuronal membranes and their synthesis is regulated by lactosylceramide N-acetylglucosaminyltransferase (LcOse2Cer-GlcNAcTr). PMID- 11874206 TI - Concentrations of different sterols in the striatum and serum of 3-nitropropionic acid-treated Wistar and Lewis rats. AB - In the present study, we examined the long-term effect of oxidative stress induced neurodegeneration on sterol concentrations in striatum and serum of Wistar and Lewis rats. Three weeks after treatment with 3-nitropropionic acid, no differences in 24S-hydroxycholesterol concentrations were observed in striatal homogenates and serum. Ratios of striatal campesterol to cholesterol were higher after 3-nitropropionic acid treatment as compared to controls of both rat strains suggesting an increased passage of this exogenous plant sterol across the blood brain-barrier. Ratios of lathosterol to cholesterol in serum and striatum were lower in treated rats as compared with controls of both rat strains. Absolute concentrations of serum and striatal cholesterol precursors and plant sterols differed between the controls of both rat strains. It was concluded that the changes observed in sterol concentrations in the striatum and serum indicate that cholesterol homeostasis may be affected during neurodegenerative processes associated with blood-brain-barrier damage. PMID- 11874207 TI - EGb761 blocks MPP+-induced lipid peroxidation in mouse corpus striatum. AB - EGb761 has been suggested to be an antioxidant and free radical scavenger. Excess generation of free radicals, leading to lipid peroxidation (LP), has been proposed to play a role in the damage to striatal neurons induced by 1-methyl-4 phenylpyridinium (MPP+). We investigated the effects of EGb761 pretreatment on MPP+ neurotoxicity. C-57 black mice were pretreated with EGb761 for 17 days at different doses (0.63, 1.25, 2.5, 5 or 10 mg/kg) followed by administration of MPP+, (0.18, 0.36 or 0.72 mg/kg). LP was analyzed in corpus striatum at 30 min, 1 h, 2 h and 24 h after MPP+ administration. Striatal dopamine content was analyzed by HPLC at the highest EGb761 dose at 2 h and 24 h after MPP+ administration. MPP+-induced LP was blocked (100%) by EGb761 (10 mg/kg). Pretreatment with EGb761 partially prevented (32%) the dopamine-depleting effect of MPP+ at 24 h. These results suggest that supplements of EGb761 may be effective at preventing MPP+ induced oxidative stress. PMID- 11874208 TI - An endogenous Na+, K+-ATPase inhibitor enhances phosphoinositide hydrolysis in neonatal but not in adult rat brain cortex. AB - The effect of an endogenous Na+, K+-ATPase inhibitor, termed endobain E, on phosphoinositide hydrolysis was studied in rat brain cortical prisms and compared with that of ouabain. As already shown for ouabain, a transient effect was obtained with endobain E; maximal accumulation of inositol phosphates induced by endobain E was 604 +/- 138% and 186 +/- 48% of basal values in neonatal and adult rats, respectively. The concentration-response plot for the interaction between endobain E and phosphoinositide turnover differed from that of ouabain, thus suggesting the involvement of distinct mechanisms. In the presence of endobain E plus ouabain at saturating concentrations, no additive effect was recorded, suggesting that both substances share at least a common step in their activation mechanism of inositol phosphates metabolism or that they enhance phosphatidylinositol 4,5-biphosphate breakdown from the same membrane precursor pool, until its exhaustion. Experiments with benzamil, a potent blocker of Na+/Ca2+ exchanger, showed that it partially and dose-dependently inhibited endobain E effect. These results indicate that the endogenous Na+, K+-ATPase inhibitor endobain E, like ouabain, is able to stimulate phosphoinositide turnover transiently during postnatal brain development. PMID- 11874209 TI - Obsessive-compulsive symptoms during treatment with olanzapine and risperidone: a prospective study of 113 patients with recent-onset schizophrenia or related disorders. AB - OBJECTIVE: To determine whether severity of obsessive-compulsive symptoms (OCS) differs during treatment with olanzapine or risperidone and to establish whether duration of antipsychotic treatment is related to severity of OCS. METHOD: We conducted a prospective study of consecutively hospitalized young patients (mean age = 22.4 years) with DSM-IV schizophrenia or related disorders (N = 113) who were treated with olanzapine or risperidone. Olanzapine or risperidone was randomly prescribed for patients who were drug-naive or were treated with typical antipsychotics before admission (N = 36). Patients who had started olanzapine (N = 39) or risperidone treatment (N = 23) prior to admission continued with that medication if they showed initial clinical response. Patients who prior to admission started olanzapine (N = 6) or risperidone (N = 9) but showed no response or suffered from adverse effects switched at admission to risperidone or olanzapine, respectively. Medical records, parents, and patients revealed information on duration of treatment and compliance with olanzapine or risperidone prior to admission. The Yale-Brown Obsessive Compulsive Scale (YBOCS) was administered at admission and 6 weeks thereafter. RESULTS: At baseline and 6 week assessments, OCS were found in about 30% of 106 evaluable cases and 15% met DSM-IV criteria for obsessive-compulsive disorder. No differences in OCS were found in the patients randomly assigned to olanzapine or risperidone. The 35 subjects treated with olanzapine at both assessments had significantly (p = .01) more severe OCS at week 6 than the 20 subjects treated with risperidone at both assessments. Duration of treatment with olanzapine was significantly (p < .01) related to severity of OCS. CONCLUSION: There are no differences in the short term propensity of olanzapine or risperidone to induce or exacerbate OCS. However, severity of OCS was associated with duration of treatment with olanzapine. PMID- 11874210 TI - Comparison between the effects of atypical and traditional antipsychotics on work status for clients in a psychiatric rehabilitation program. AB - BACKGROUND: Although many studies have compared the impact of atypical antipsychotics with that of traditional antipsychotics on psychiatric symptoms, few have compared the impact on work status, especially in the context of best practices psychiatric rehabilitation. METHOD: A cross-sectional design examined symptom and employment status for 82 clients with DSM-IV schizophrenia-spectrum disorders who had attended a psychiatric rehabilitation program for a mean of 5 years. Using chart review and client interviews, we examined the relationship between type of antipsychotic prescribed and symptom and work status in 59 clients prescribed an atypical antipsychotic (olanzapine or risperidone) for a mean of 20 months and 23 clients prescribed a traditional antipsychotic for a mean of 75 months. Measures included the Positive and Negative Syndrome Scale and 2 work status measures: an 8-point employment status scale (the Work Placement Scale) and percentage of clients working in independent employment. RESULTS: The atypical group had significantly fewer symptoms of cognitive impairment and hostility/excitement than the traditional group (p < .05). However, self-reported adverse events were similar in the 2 medication groups, and the 2 groups did not differ significantly on work status. Less severe negative, cognitive, and hostility/excitement symptoms were associated with more independent employment status. CONCLUSION: For long-term clients in a psychiatric rehabilitation program, type of medication prescribed was associated with better symptom control but not better work status. The association between symptoms and work status, however, may suggest an indirect link favoring atypical antipsychotics for achieving paid employment. PMID- 11874211 TI - No weight gain among elderly schizophrenia patients after 1 year of risperidone treatment. AB - OBJECTIVE: Weight gain has been reported in younger patients treated with most atypical neuroleptics. The goal of this study was to examine whether elderly schizophrenic patients gain weight while being treated with risperidone. METHOD: Data are from an international multicenter, open-label study of 180 elderly (69 men, 111 women), chronically ill, psychotic patients (meeting DSM-III-R criteria for schizophrenia or schizophreniform disorder; median age = 72 years [range, 54 89 years]), 97 of whom completed the 12-month study. At endpoint, the mean dose of risperidone was 3.7 mg/day. Patients were weighed at baseline and at endpoint. RESULTS: There was no significant weight gain in patients who completed the trial (N = 96) or in those who did not complete the entire trial (N = 31). CONCLUSION: The results suggest that risperidone treatment is not associated with weight gain among elderly persons with chronic psychosis. PMID- 11874212 TI - Demographic and clinical characteristics of individuals in a bipolar disorder case registry. AB - BACKGROUND: The goal of this analysis was to characterize a cohort of 3000 persons who self-identified as having bipolar disorder by demographic, clinical, and treatment characteristics and to document the burden that this disorder imposed on their lives. METHOD: The Stanley Center Bipolar Disorder Registry used a variety of recruitment methods to reach people with bipolar disorder. The cohort included those currently in treatment and those active in support groups. Registrants completed an interviewer-administered questionnaire to obtain information on demographic characteristics, clinical history, and treatment history. RESULTS: The median age of the 2839 patients who were analyzed was 40.1 years, 64.5% were women, and over 90% were white. The median age at onset was 17.5 years, and the mean was 19.8 years. Despite the fact that over 60% completed at least some college and 30% completed college, 64% were currently unemployed. The patients' family histories point to a high prevalence of mental disorder in the families, especially mood disorders. Patients were concurrently taking multiple medications, and more than one third were taking at least 3 types of psychotropic medications. This pattern of pharmacotherapy was consistent with participants' overall mood ratings, which demonstrated how unusual it was for them to be symptom-free over a 6-month period. CONCLUSION: Our present findings point to the chronicity and severity of bipolar disorder as experienced in the community. We still need to develop better interventions, ensure access to care consistent with current consensus guidelines, and initiate care as early as possible in the course of the condition. PMID- 11874213 TI - Current comorbidity of psychiatric disorders among DSM-IV major depressive disorder patients in psychiatric care in the Vantaa Depression Study. AB - BACKGROUND: While numerous studies have documented the high comorbidity of major depressive disorder (MDD) with individual mental disorders, no published study has reported overall current comorbidity with all Axis I and II disorders among psychiatric patients with MDD, nor systematically investigated variations in current comorbidity by sociodemographic factors, inpatient versus outpatient status, and number of lifetime depressive episodes. METHOD: Psychiatric outpatients and inpatients in Vantaa, Finland, were prospectively screened for an episode of DSM-IV MDD, and 269 patients with a new episode of MDD were enrolled in the Vantaa Depression MDD Cohort Study. Axis I and II comorbidity was assessed via semistructured Schedules for Clinical Assessment in Neuropsychiatry, version 2.0, and Structured Clinical Interview for DSM-II-R personality disorders interviews. RESULTS: The great majority (79%) of patients with MDD suffered from 1 or more current comorbid mental disorders, including anxiety disorder (57%), alcohol use disorder (25%), and personality disorder (44%). Several anxiety disorders were associated with specific Axis II clusters, and panic disorder with agoraphobia was associated with inpatient status. The prevalence of personality disorders varied with inpatient versus outpatient status, number of lifetime depressive episodes, and type of residential area, and the prevalence of substance use disorders varied with gender and inpatient versus outpatient status. CONCLUSION: Most psychiatric patients with MDD have at least 1 current comorbid disorder. Comorbid disorders are associated not only with other comorbid disorders, but also with sociodemographic factors, inpatient versus outpatient status, and lifetime number of depressive episodes. The influence of these variations on current comorbidity patterns among MDD patients needs to be taken account of in treatment facilities. PMID- 11874214 TI - Hepatotoxicity associated with the new antidepressants. AB - BACKGROUND: Safety profiles of classical and new antidepressants are well established. Hepatotoxicity is known to occur. Recently, several cases of severe hepatic injury associated with the new antidepressants have been reported, prompting us to quantify this risk. METHOD: To estimate the cumulative incidence of hepatic adverse reactions associated with antidepressants, we used cases of hepatic damage collected via spontaneous reporting and included in the Spanish Pharmacovigilance System database; for exposure, we have used data from drug sales to the Spanish National Health System. RESULTS: The estimated reported incidence did not show major differences for the antidepressants studied, ranging from 1.28 cases per 100,000 patient-years for sertraline to 4.00 for clomipramine, except for nefazodone, which was the agent that had the highest incidence with 28.96 cases per 100,000 patient-years. CONCLUSION: The reported incidence of hepatic adverse reactions to nefazodone seems to be higher than that estimated so far. Given the high prevalence of depression and the widespread use of antidepressants, physicians should be alert to the possibility that these medications cause hepatitis and consider early discontinuation of an antidepressant if the condition is suspected. PMID- 11874215 TI - Behavior therapy for obsessive-compulsive disorder guided by a computer or by a clinician compared with relaxation as a control. AB - BACKGROUND: The demand for effective behavior therapy for obsessive-compulsive disorder (OCD) by exposure and ritual prevention exceeds its supply by trained therapists. A computer-guided behavior therapy self-help system (BT STEPS) was created that patients access by telephone from home via interactive voice response technology. This study compared the value of computer-guided behavior therapy value with that of clinician-guided behavior therapy and systematic relaxation as a control treatment. METHOD: After screening by a clinician, 218 patients with DSM-IV OCD at 8 North American sites were randomly assigned to 10 weeks of behavior therapy treatment guided by (1) a computer accessed by telephone and a user workbook (N = 74) or (2) a behavior therapist (N = 69) or (3) systematic relaxation guided by an audiotape and manual (N = 75). RESULTS: By week 10, in an intent-to-treat analysis, mean change in score on the Yale-Brown Obsessive Compulsive Scale was significantly greater in clinician-guided behavior therapy (8.0) than in computer-guided (5.6), and changes in scores with both clinician-guided and computer-guided behavior therapy were significantly greater than with relaxation (1.7), which was ineffective. Similarly, the percentage of responders on the Clinical Global Impressions scale was significantly (p < .05) greater with clinician-guided (60%) than computer-guided behavior therapy (38%), and both were significantly greater than with relaxation (14%). Clinician-guided was superior to computer-guided behavior therapy overall, but not when patients completed at least 1 self-exposure session (N = 36 [65%]). At endpoint, patients were more satisfied with either behavior therapy group than with relaxation. Patients assigned to computer-guided behavior therapy improved more the longer they spent telephoning the computer (mostly outside usual office hours) and doing self-exposure. They improved slightly further by week 26 follow-up, unlike the other 2 groups. CONCLUSION: For OCD, computer-guided behavior therapy was effective, although clinician-guided behavior therapy was even more effective. Systematic relaxation was ineffective. Computer-guided behavior therapy can be a helpful first step in treating patients with OCD when clinician-guided behavior therapy is unavailable. PMID- 11874216 TI - Stabilization of mood from below versus above baseline in bipolar disorder: a new nomenclature. AB - Management of bipolar disorder has traditionally emphasized the acute treatment of mania. Although acute treatment of mania is a critical aspect of care, this emphasis has tended to overshadow other important phases of bipolar disorder, such as depression, hypomania, and subsyndromal symptoms. We offer a reconceptualization of bipolar disorder that highlights unmet needs and the importance of differential spectra of efficacy. In this reconceptualization, bipolar disorder can be viewed as an aberration of mood, behavior, and cognition from baseline (euthymia). "Below baseline" is characterized by depression and subsyndromal depression. "Above baseline" is characterized by mania, mixed states, hypomania, and subsyndromal mood elevation. In contrast to the treatment options for mania, the options for depression are limited. This new nomenclature emphasizes the need to develop mood stabilizers that possess the ability to stabilize mood "from below baseline," either alone or in combination with other agents. In this article, the treatment options for bipolar disorder, with a focus on depression and rapid cycling, are discussed according to this new conceptualization of management from below and above baseline. PMID- 11874217 TI - Predictors of response to pharmacotherapy in social anxiety disorder: an analysis of 3 placebo-controlled paroxetine trials. AB - BACKGROUND: There is increasing evidence that patients with social anxiety disorder (social phobia) respond to treatment with selective serotonin reuptake inhibitors (SSRIs). Response rates to SSRIs in social anxiety disorder have ranged from at least 50% in controlled trials to up to 80% in open trials. To date, however, there has been little information available about predictors of response to treatment in this disorder. METHOD: Data from 3 placebo-controlled multicenter trials of paroxetine in DSM-IV social anxiety disorder (N = 829) were analyzed using logistic regression to determine predictors of response. Demographic (age, sex), physiologic (baseline heart rate, baseline mean arterial pressure), clinical (baseline social anxiety symptom severity, baseline disability, duration of illness), and trial variables (paroxetine dose, treatment duration) were included. RESULTS: Only duration of treatment was a statistically significant predictor of treatment response. Further analysis demonstrated that, in paroxetine-treated patients in particular, many nonresponders at week 8 (46/166; 27.7%) were responders at week 12. CONCLUSION: These data demonstrate that paroxetine is a reasonable choice of treatment in a broad spectrum of patients with social anxiety disorder. An optimal trial of medication should continue beyond 8 weeks. PMID- 11874219 TI - Organic hypomania secondary to sibutramine-citalopram interaction. PMID- 11874218 TI - Duration of therapy and health care costs of fluoxetine, paroxetine, and sertraline in 6 health plans. AB - BACKGROUND: Previous studies comparing fluoxetine, paroxetine, and sertraline, the 3 most common selective serotonin reuptake inhibitors (SSRIs), in naturalistic settings have produced conflicting results. With this study, we provide new evidence as to the similarities and differences among these SSRI therapies with respect to the duration of use and health care costs. METHOD: Data from 6 health maintenance organizations were used to identify patients with new onset major depression. number of days with filled prescriptions, and total health care and depression-related costs. The sample consisted of 1771 patients given initial prescriptions for sertraline (N = 386), fluoxetine (N = 840), or paroxetine (N = 545) in the period from July 1, 1994, to March 31, 1997. Analyses included Cox proportional hazards models (for duration of initial therapy) and ordinary least squares regression (for cost). RESULTS: Patients who initiated therapy with fluoxetine were more likely to have a later interruption of therapy than patients who initiated therapy with sertraline (p = .03) and paroxetine (p = .001). Total 1-year costs did not differ statistically between the treatment groups, but 1-year depression-related costs were significantly lower for patients who initiated therapy with sertraline or paroxetine than for those who initiated therapy with fluoxetine ($332 less for sertraline, 95% confidence interval [CI] = $125 to $562; $339 less for paroxetine, 95% CI = $144 to $416). LIMITATIONS: A limitation of this observational study, as well as of observational studies in general, is that unobserved characteristics of the patients may lead to biased estimates of the impact of treatment on adherence or cost, even with controls for observed characteristics. CONCLUSION: We found no significant differences in total health care costs among the 3 SSRIs, but noted significant differences in depression-related costs (the costs of fluoxetine are greater than those of sertraline and paroxetine). Importantly, there was no relationship between treatment interruption and increased health care or depression-related costs, in contrast to the findings of some, but not all, prior studies. PMID- 11874220 TI - Risperidone-induced Pisa syndrome. PMID- 11874221 TI - Interferon-induced depression treated with citalopram. PMID- 11874222 TI - Clozapine and tardive dyskinesia. PMID- 11874223 TI - Role of receptor binding profiles in antidepressant-induced sexual dysfunction. PMID- 11874224 TI - Olanzapine for the treatment of monosymptomatic hypochondriacal psychosis. PMID- 11874225 TI - Safety and tolerability of a rapidly escalating dose-loading regimen for risperidone. PMID- 11874226 TI - Antipsychotic polypharmacy: squandering precious resources? PMID- 11874227 TI - What role do atypical antipsychotic drugs have in treatment-resistant depression? AB - Despite significant advances in the treatment of depression, many patients fail to respond to treatment with adequate dose and duration. Multiple therapeutic approaches are available for the treatment of patients not responding to standard antidepressant medication. These include switching medication or combination and augmentation strategies. A substantial number of patients do not respond to multiple treatment trials. These patients suffer from treatment-resistant depression (TRD) and represent a challenge to the treating physician. There have been a growing number of reports on the use of atypical antipsychotics as augmenting agents in nonpsychotic TRD. Second-generation antipsychotics are less likely to provoke parkinsonian side effects. It has also been reported that these agents produce lower rates of tardive movement disorders than traditional neuroleptics. Furthermore, second-generation antipsychotics are serotonin-2A/2C antagonists, possibly allowing them to improve the efficacy and some aspects of the side effect profile of selective serotonin reuptake inhibitors (SSRIs). Weight gain and sedation are likely to be the most common adverse events of such combined therapy. In a recent controlled clinical trial, the atypical antipsychotic olanzapine was combined with fluoxetine therapy in an 8-week, double-blind clinical trial in patients with TRD. This combination drug therapy demonstrated clinical efficacy on several rating scales and showed rapid onset of action. Although more studies will be required to confirm and extend these findings, the results suggest that there may be a clinical benefit to combining atypical antipsychotics with SSRIs in nonpsychotic TRD. PMID- 11874228 TI - A randomized school-based jumping intervention confers site and maturity-specific benefits on bone structural properties in girls: a hip structural analysis study. AB - We compared 7-month changes in bone structural properties in pre- and early pubertal girls randomized to exercise intervention (10-minute, 3 times per week, jumping program) or control groups. Girls were classified as prepubertal (PRE; Tanner breast stage 1; n = 43 for intervention [I] and n = 25 for control [C]) or early-pubertal (EARLY; Tanner stages 2 and 3; n = 43 for I and n = 63 for C). Mean +/- SD age was 10.0 +/- 0.6 and 10.5 +/- 0.6 for the PRE and EARLY groups, respectively. Proximal femur scans were analyzed using a hip structural analysis (HSA) program to assess bone mineral density (BMD), subperiosteal width, and cross-sectional area and to estimate cortical thickness, endosteal diameter, and section modulus at the femoral neck (FN), intertrochanter (IT), and femoral shaft (FS) regions. There were no differences between intervention and control groups for baseline height, weight, calcium intake, or physical activity or for change over 7 months (p > 0.05). We used analysis of covariance (ANCOVA) to examine group differences in changes of bone structure, adjusting for baseline weight, height change, Tanner breast stage, and physical activity. There were no differences in change for bone structure in the PRE girls. The more mature girls (EARLY) in the intervention group showed significantly greater gains in FN (+2.6%, p = 0.03) and IT (+1.7%, p = 0.02) BMD. Underpinning these changes were increased bone cross-sectional area and reduced endosteal expansion. Changes in subperiosteal dimensions did not differ. Structural changes improved section modulus (bending strength) at the FN (+4.0%, p = 0.04), but not at the IT region. There were no differences at the primarily cortical FS. These data provide insight into geometric changes that underpin exercise-associated gain in bone strength in early-pubertal girls. PMID- 11874229 TI - An exercise in geometry. PMID- 11874230 TI - Imaging of Erdheim-Chester disease. PMID- 11874231 TI - Two new single-nucleotide polymorphisms in the COL1A1 upstream regulatory region and their relationship to bone mineral density. AB - Single-nucleotide polymorphisms (SNPs) in regulatory regions of candidate genes may determine variability in bone mineral density (BMD) because they may be responsible for differences in levels of a gene product in response to external signals. Under this hypothesis, we scanned an 800-base pair (bp) region within the COL1A1 promoter, known to harbor cis elements important for in vivo expression, and we found two new polymorphisms: -1663indelT and -1997 G/T. The G to T transversion at -1997 was associated with lumbar spine BMD (p = 0.015) when tested in a cohort of 256 postmenopausal women after adjusting by age, body weight, and years since menopause; a lower degree of association was detected also for femoral neck BMD in a subgroup of 146 women in univariate analysis and after adjusting by age (p = 0.044). The polymorphism -1663indelT, which corresponds to a deletion of a T in a tract of eight T residues (-1670 to -1663), did not show significant association with BMD. Interestingly, -1663indelT is in strong linkage disequilibrium (LD) with the previously described Sp1 polymorphism of intron 1, which in this study did not show association with BMD either. Significant interaction between -1997 G/T and -1663indelT (p = 0.019), and between -1997 G/T and Sp1 (p = 0.045) was observed also. Individuals heterozygous for the three polymorphisms showed the highest mean BMD value. Gel retardation assays showed that oligonucleotides containing either the -1663 or the -1997 polymorphic sites specifically bind primary osteoblast nuclear proteins. We named these binding sites as PCOL1 and PCOL2, respectively. In summary, this study describes two new SNPs in the COL1A1 promoter, which may affect bone mass determination. PMID- 11874232 TI - Overexpressed LIM mineralization proteins do not require LIM domains to induce bone. AB - Rat LIM mineralization protein 1 (LMP-1, an LIM domain protein) mediates bone morphogenetic protein 6 (BMP-6) induction of bone nodule formation in fetal rat calvarial osteoblast (ROB) cultures. We have isolated the complementary DNA (cDNA) for the human homologue of LMP-1 from an adult human heart cDNA library and showed that when overexpressed it is osteoinductive in the same culture system. The recently revised cDNA sequence of Enigma, the protein product of which binds to the insulin receptor and the tyrosine kinase receptor ret, now matches the nucleotide sequence of human LMP-1 (hLMP-1). A truncated, 223 amino acid (AA) LMP-1(t) protein has identical effects as the full-length protein, despite the deletion of the LIM domains. Two splice variants of human LMP-1 have been detected. Human LMP-2 has a 119-base pair (bp) deletion between bp 325 and 444 and a 17-bp insertion at bp 444. The resulting derived protein contains 423 AA with the LIM domains intact and does not induce bone formation when overexpressed in ROB cultures. Human LMP-3 has the same 17 nucleotide insertion at bp 444, resulting in a shift in the reading frame that causes a stop codon to occur at bp 505-507. The resulting 153 AA protein does not have the LIM domains, but overexpression of hLMP-3 induces bone formation in osteoblast cultures. These findings suggest that the LIM domains are not required for LMPs to induce bone formation. In addition, a small region (36 AA) of the LMP-1 protein may be required for bone formation. PMID- 11874233 TI - The secreted protein thrombospondin 2 is an autocrine inhibitor of marrow stromal cell proliferation. AB - Marrow stromal cells (MSCs) are obtained in increased number from mice in which the thrombospondin 2 (TSP2) gene is disrupted, and these cells show increased DNA synthesis in vitro. To examine more closely the role of TSP2 in the physiology and osteogenic differentiation of MSCs, an in-depth characterization of TSP2-null MSCs was conducted. Determination of TSP2 protein content by Western analysis and RNA levels by reverse-transcription polymerase chain reaction (RT-PCR) indicated that MSCs are the primary source of TSP2 in the marrow and secrete abundant TSP2 into culture medium. Morphologically, the TSP2-null and wild-type (WT) cell populations were similar and by flow cytometry contained equivalent numbers of CD44+, Mac1+, intercellular adhesion molecule-1 (ICAM-1+), and ScaI+ cells. TSP2 null cells showed delayed mineralization associated with an increased rate of proliferation. Consistent with this finding, there was a decrease in expression of collagen and osteocalcin RNA by TSP2-null MSCs on day 7 and increased osteopontin expression on day 7 and day 14. In add-back experiments, recombinant TSP2 produced a dose-dependent decrease in proliferation. This reduction was associated with an accumulation of TSP2-treated cells in the G1 phase of the cell cycle and did not result from an increase in apoptosis. When TSP2 treatment was terminated, the cell population reentered the S phase. We conclude that the increased endosteal bone formation observed in TSP2-null mice results primarily from the failure of TSP2 to regulate locally MSC cell cycle progression. PMID- 11874234 TI - The uptake of lipoprotein-borne phylloquinone (vitamin K1) by osteoblasts and osteoblast-like cells: role of heparan sulfate proteoglycans and apolipoprotein E. AB - Vitamin K is essential for the gamma-carboxylation of Gla-containing bone proteins such as osteocalcin and a suboptimal vitamin K status has been linked to osteoporosis but nothing is known of how the lipoprotein-borne vitamin accesses the bone matrix. We have studied the mechanism of transport of lipoproteins labeled with [3H]-phylloquinone (vitamin K1 [K1]) into osteoblasts using both tumor-derived cell lines and normal osteoblast-rich cell populations. We also investigated the effect of heparin in this model since long-term heparin treatment causes osteopenia and the anticoagulant is known to impair normal lipoprotein metabolism. Heparinase treatment, which removes heparan sulfate proteoglycans (HSPG), reduced uptake of [3H]-K1 from triglyceride-rich lipoproteins (TRL) and low-density lipoproteins (LDL). The effect of heparin in this model was complex depending on cell type, concentration, and time but, overall, the results were consistent with an inhibition of vitamin K uptake by osteoblasts. Anti-apolipoprotein E (apoE) antiserum reduced uptake of TRL-[3H]-K1 by 55 +/- 4% and LDL-[3H]-K1 uptake by 35 +/- 2%. Exogenous apoE4 increased uptake of TRL-[3H]-K1 by 90 +/- 1% compared with 53 +/- 11% for apoE3 and 52 +/- 5% for apoE2. Our findings show that HSPG on the cell surface and apoE in the lipoprotein particles contribute to lipoprotein-K1 uptake by osteoblasts as is known for lipoprotein uptake by hepatocytes. This mechanism is significant in view of the epidemiological association of both undercarboxylation of osteocalcin and the presence of an apo epsilon4 allele with increased fracture risk and reduced bone mineral density (BMD). The inhibition by heparin of lipoprotein mediated carriage of vitamin K and possibly other lipids to bone may provide a basis for the future understanding of heparin-induced osteoporosis. PMID- 11874235 TI - Evidence that peroxynitrite affects human osteoblast proliferation and differentiation. AB - Peroxynitrite (PN), a nitric oxide (NO*)-derived anion, has been associated with NO* damage in various cell types. We examined the effects of adding PN to cultured human osteoblast-like (hOB) cells obtained after hip arthroplasty. Exposure to PN (0.1-0.4 mM) decreased both hOB proliferation and differentiation, measured by [3H]thymidine uptake and alkaline phosphatase production, respectively. Incubation with 3-morpholinosydnonimine (SIN-1; 0.25-1 mM), an NO* and O2- donor that leads to PN release, also reduced both hOB proliferation and differentiation. Coincubation with both superoxide dismutase (SOD; 100 U/ml) and catalase (CAT; 50 U/ml), rendering SIN-1 a pure NO* donor, reversed its effects on hOB proliferation and differentiation. However, SIN-1-induced NO* production, measured by nitrite release to the hOB medium, was not altered by cotreatment with SOD and CAT. Expression of nitrotyrosine by hOB, a marker of PN action, was significantly increased after SIN-1 addition, as compared with untreated cells, as revealed by Western blot analysis. Interleukin-1alpha (IL-1alpha) and interferon gamma (IFN-gamma) but not tumor necrosis factor alpha (TNF-alpha) also significantly increased nitrotyrosine expression in these cells. These data show that PN is at least partially responsible for osteoblast derangement by NO* and that cytokines released during inflammatory arthropathies can induce PN production in hOB cells. PMID- 11874236 TI - Thyroid hormones promote chondrocyte differentiation in mouse ATDC5 cells and stimulate endochondral ossification in fetal mouse tibias through iodothyronine deiodinases in the growth plate. AB - Thyroid hormones (THs), 3,3',5-triiodo-L-thyronine (T3) and L-thyroxine (T4), are important for the normal development of the growth plate (GP); congenital TH deficiency leads to severe dwarfism. In mouse chondrogenic cell line, ATDC5, T3 enhanced differentiation and increased Alizarin red staining, but did not affect Alcian blue staining. In organ-cultured mouse tibias, THs stimulated the cartilage growth, especially in the hypertrophic zone. Interestingly, T4 was as equally potent as T3 in organ-cultured tibias, which suggests that T4 is metabolized locally to T3, because T4 is a prohormone and must be converted to T3 for its activity. Two enzymes catalyze the conversion; type I deiodinase (D1) and type II deiodinase (D2). D1 has a ubiquitous distribution and D2, with a high affinity for T4, is present where the maintenance of intracellular T3 concentration is critical. Messenger RNAs (mRNAs) for D1 and D2 were detected in neonatal mouse tibias and ATDC5 cells. The enzyme activity was unaffected by the D1 inhibitor 6-propyl-2-thiouracil, suggesting that D2 mainly catalyzes the reaction. D2 mRNA was detected in differentiated ATDC5 cells. In organ-cultured mouse tibias, D2 activity was greater at later stages. In contrast, thyroid hormone receptors (TRs) were expressed in neonatal mouse tibias and ATDC5 cells, but their expression levels in ATDC5 cells were stable throughout the culture periods. Therefore, increased T3 production at later stages by D2 is likely to contribute to the preferential effects of THs in the terminal differentiation of GP. This article is the first to show that T4 is activated locally in GP and enhances the understanding of TH effects in GP. PMID- 11874237 TI - Nuclear localization of type I parathyroid hormone/parathyroid hormone-related protein receptors in deer antler osteoclasts: evidence for parathyroid hormone related protein and receptor activator of NF-kappaB-dependent effects on osteoclast formation in regenerating mammalian bone. AB - Parathyroid hormone-related protein (PTHrP) is not required for osteoclastogenesis during embryonic development; however, after birth it has been shown to regulate osteoclast formation during tooth eruption. Our study explores the hypothesis that PTHrP also may regulate osteoclast differentiation in the regenerating skeletal tissues of deer antlers, bones capable of complete regeneration. Osteoclast-like multinucleated cells (MNCs) formed spontaneously in micromass cultures derived from antler cartilage and these cells had the phenotypic characteristics of osteoclasts. PTHrP and receptor activator of NF kappaB ligand (RANKL) stimulated antler osteoclast formation although the effect of RANKL was less marked than that of PTHrP. The addition of osteoprotegerin (OPG) only partially decreased (by approximately 65%) the number of osteoclasts in PTHrP-treated cultures. To determine whether PTHrP also potentially could have direct effects on antler osteoclasts, we studied, by confocal microscopy, the expression of the type I PTH/PTHrP receptor (PTH1R) in MNCs cultured on glass and found the receptor protein to have a nuclear localization. In situ hybridization showed that antler MNCs also expressed PTH1R and PTHrP messenger RNAs (mRNAs). PTHrP was immunolocalized in MNCs cultured on glass but was undetectable in cells resorbing a dentine substrate. In tissue sections of antler cartilage, PTHrP and PTH1R were expressed in vitronectin receptor-positive (VNR+) osteoclast-like cells localized in the perivascular stroma. Thus, these data show that PTHrP plays a role in the regulation of osteoclast differentiation in regenerating skeletal tissues and that PTHrP can have effects on osteoclastogenesis that are independent of RANKL synthesis. Ours is the first study to describe the expression of the type I PTH/PTHrP receptor in mammalian osteoclasts at a protein and mRNA level, which indicates that PTHrP also may have a direct effect on osteoclasts. This also is the first study to show a nuclear localization of the PTHIR in cells of the osteoclast lineage, although the functional significance of this observation has yet to be established. PMID- 11874238 TI - Nuclear magnetic resonance spin-spin relaxation of the crystals of bone, dental enamel, and synthetic hydroxyapatites. AB - Studies of the apatitic crystals of bone and enamel by a variety of spectroscopic techniques have established clearly that their chemical composition, short-range order, and physical chemical reactivity are distinctly different from those of pure hydroxyapatite. Moreover, these characteristics change with aging and maturation of the bone and enamel crystals. Phosphorus-31 solid state nuclear magnetic resonance (NMR) spin-spin relaxation studies were carried out on bovine bone and dental enamel crystals of different ages and the data were compared with those obtained from pure and carbonated hydroxyapatites. By measuring the 31P Hahn spin echo amplitude as a function of echo time, Van Vleck second moments (expansion coefficients describing the homonuclear dipolar line shape) were obtained and analyzed in terms of the number density of phosphorus nuclei. 31P magnetization prepared by a 90 degree pulse or by proton-phosphorus cross polarization (CP) yielded different second moments and experienced different degrees of proton spin-spin coupling, suggesting that these two preparation methods sample different regions, possibly the interior and the surface, respectively, of bone mineral crystals. Distinct differences were found between the biological apatites and the synthetic hydroxyapatites and as a function of the age and maturity of the biological apatites. The data provide evidence that a significant fraction of the protonated phosphates (HPO4(-2)) are located on the surfaces of the biological crystals, and the concentration of unprotonated phosphates (PO4(-3)) within the apatitic lattice is elevated with respect to the surface. The total concentration of the surface HPO4(-2) groups is higher in the younger, less mature biological crystals. PMID- 11874239 TI - A comparison of type I collagen, fibronectin, and vitronectin in supporting adhesion of mechanically strained osteoblasts. AB - We used an adhesion assay for cells cultured under high dynamic strain to measure human osteoblast-like HOS cell adherence to immobilized type I collagen, fibronectin, and vitronectin. These conditions were designed to model the increased forces present at unstable fractures or loose joint prostheses. At a constant, low protein-coating density (1000 molecules/microm2) and 20% cyclic strain for 24 h, type I collagen, fibronectin, and vitronectin supported 24.6 +/- 2%, 16.7 +/- 3%, and 1.1 +/- 1% adherence, respectively, which paralleled the relative number of integrin-binding sites in each protein. Thus, when the number of available binding sites was limited, strain resistance was proportional to the number of integrin-ligand interactions. In contrast, at high protein-coating densities (> or = 2,500 molecules/microm2), vitronectin supported greater adherence (45.7 +/- 2%) when compared with type I collagen (37 +/- 2%) or fibronectin (34.8 +/- 2%) and directed constitutive expression of osteopontin (OPN), which suggested that there exist discrete signals on vitronectin receptor occupancy that promoted cell adherence and survival under strain. Integrin mediated binding was necessary for resistance to strain, as evidenced by the low levels of strain resistance observed when cells were adherent in a nonintegrin dependent manner. These findings support the utilization of at least two distinct mechanisms (i.e., tensegrity and integrin-mediated signal transduction) by HOS cells to remain adherent and viable on exposure to mechanical forces. PMID- 11874240 TI - Noninvasive loading of the murine tibia: an in vivo model for the study of mechanotransduction. AB - Transgenic and knockout mice present a unique opportunity to study mechanotransduction pathways in vivo, but the difficulty inherent with applying externally controlled loads to the small mouse skeleton has hampered this approach. We have developed a novel device that enables the noninvasive application of controlled mechanical loads to the murine tibia. Calibration of tissue strains induced by the device indicated that the normal strain environment was repeatable across loading bouts. Two in vivo studies were performed to show the usefulness of the device. Using C57Bl/6J mice, we found that dynamic but not static loading increased cortical bone area. This result is consistent with previous models of bone adaptation, and the lack of adaptation induced by static loading serves as a negative control for the device. In a preliminary study, transgenic mice selectively overexpressing insulin-like growth factor 1 (IGF-1) in osteoblasts underwent a low-magnitude loading regimen. Periosteal bone formation was elevated 5-fold in the IGF-1-overexpressing mice but was not elevated in wild-type littermates, showing the potential for synergism between mechanical loading and selected factors. Based on these data, we anticipate that the murine tibia-loading device will enhance assessment of mechanotransduction pathways in vivo and, as a result, has the potential to facilitate novel gene discovery and optimization of synergies between drug therapies and mechanical loading. PMID- 11874241 TI - Regulation of fibroblast growth factor 2 and fibroblast growth factor receptors by transforming growth factor beta in human osteoblastic MG-63 cells. AB - Fibroblast growth factor 2 (FGF-2) and its receptors (FGFRs) are important regulators of bone cell function. Although FGF-2 is a major modulator of bone cell function, its expression and regulation in human osteoblasts have not been investigated. We examined FGF-2 messenger RNA (mRNA) expression and regulation in the human osteosarcoma MG-63 cells. Northern analysis revealed that MG-63 cells expressed FGF-2 mRNA transcripts of 7, 4, 2.2, and 1.3 kilobases (kb). In the absence of serum, treatment with transforming growth factor beta (TGF-beta; 0.1 10 ng/ml) increased all FGF-2 mRNA transcripts. Maximal increase was seen with 1 ng/ml of TGF-beta. TGF-beta increased FGF-2 mRNA expression within 2 h and this was sustained for 24 h. Phorbal myristate acetate (PMA; 1 microM) also increased FGF-2 mRNA at 6 h. Time course studies showed that TGF-beta did not significantly alter FGFR1 or FGFR2 mRNA expression in MG-63 cells. Western blotting with anti human FGF-2 revealed that MG-63 cells synthesize three isoforms of FGF-2 protein of approximately 18, 22/23, and 24 kDa, which were increased after either 6 h or 24 h of treatment with TGF-beta. Increased FGF-2 mRNA and protein expression in response to TGF-beta was markedly reduced by the protein kinase A (PKA) inhibitor H-89. Immunogold labeling of MG-63 cells treated with TGF-beta showed increased labeling for FGF-2 and FGFR2 in the nuclei. In contrast, TGF-beta treatment significantly decreased FGFR1 labeling in the nuclei. These data show that TGF beta regulates FGF-2 gene expression in human osteosarcoma cells. Furthermore, TGF-beta modulates the cellular localization of FGF-2 and its receptors. PMID- 11874242 TI - Differential temporal expression of members of the transforming growth factor beta superfamily during murine fracture healing. AB - Fracture healing is a unique postnatal repair process in which the events of endochondral and intramembranous bone formation follow a definable temporal sequence. The temporal patterns of messenger RNA (mRNA) expression for members of the transforming growth factor beta (TGF-beta) superfamily were examined over a 28-day period of fracture healing in mouse tibias. Bone morphogenetic protein 2 (BMP-2) and growth and differentiation factor 8 (GDF8) showed maximal expression on day 1 after fracture, suggesting their roles as early response genes in the cascade of healing events. Restricted expression of GDF8 to day 1, in light of its known actions as a negative regulator of skeletal muscle growth, suggests that it may similarly regulate cell differentiation early in the fracture healing process. GDF5, TGF-beta2, and TGF-beta3 showed maximal expression on day 7, when type II collagen expression peaked during cartilage formation. In contrast, BMP 3, BMP-4, BMP-7, and BMP-8 showed a restricted period of expression from day 14 through day 21, when the resorption of calcified cartilage and osteoblastic recruitment were most active. TGF-beta1, BMP-5 and BMP-6, and GDF10 were constitutively expressed from day 3 to day 21. However, during the same time period, GDF3, GDF6, and GDF9 could not be detected, and GDF1 was expressed at extremely low levels. These findings suggest that several members of the TGF-beta superfamily are actively involved in fracture healing and although they are closely related both structurally and functionally, each has a distinct temporal expression pattern and potentially unique role in fracture healing. PMID- 11874243 TI - A randomized trial of nasal spray salmon calcitonin in men with idiopathic osteoporosis: effects on bone mineral density and bone markers. AB - In a 12-month randomized, double-blind, placebo-controlled trial, we have studied the effects of intranasal salmon calcitonin (SCT) on bone mineral density (BMD) and biochemical markers of bone turnover. Twenty-eight men with idiopathic osteoporosis aged 27-74 years (mean, 52.4 years) were randomized to receive either nasal SCT (200 IU) or a nasal placebo daily for a period of 1 year. All the men received a daily supplement of 0.5 g of calcium. The men who received SCT had a mean (+/-SEM) increase in BMD of 7.1 +/- 1.7% at the lumbar spine. In contrast, the men who received the placebo had an increase of 2.4 +/- 1.5% (p > 0.05) for the comparison with baseline. The increase in lumbar BMD in the calcitonin group was significantly greater than that in the placebo group (p < 0.05). There were no significant changes in the femoral neck, trochanter, or Ward's triangle relative to both baseline and placebo after 12 months. Treatment with nasal SCT resulted in a significantly pronounced suppression of bone resorption markers (urinary deoxypyridinoline [DPD], type I cross-linked N telopeptide [NTX], and type I cross-linked C-telopeptide [CTX]) and to a lesser extent in bone formation markers (serum bone-specific alkaline phosphatase [BALP], osteocalcin [OC], serum C-terminal procollagen type I extension peptides [PICP], and serum N-termnal procollagen type I extension peptides [PINP]), whereas the placebo did not. Therapy was tolerated well and there were no treatment-related adverse events. We conclude that intranasal SCT (200 IU daily) is safe and effective in increasing lumbar BMD and reducing bone turnover in men with idiopathic osteoporosis. PMID- 11874244 TI - Does hormone-replacement therapy prevent fractures in early postmenopausal women? AB - The purpose of this population-based prospective cohort study was to examine the effect of hormone-replacement therapy (HRT) on the risk of fractures. The study population consisted of 7217 postmenopausal women aged 47-56 years (mean, 53.3 years) at baseline from data taken from the Kuopio Osteoporosis Risk Factor and Prevention Study (OSTPRE) in Finland. We compared fracture incidences between HRT users and nonusers. A total of 679 (9.4%) women recorded validated fractures during the 5-year follow-up. Of these, 268 (39%) women had a distal forearm fracture. Two thousand six hundred seventy women (37%) had used HRT >6 months during the follow-up--one-half of them continuously. The relative risk, estimated as hazard ratio with Cox regression, was 0.69 (95% CI, 0.58-0.82) for any fracture and 0.49 (0.36-0.66) for distal forearm fracture among HRT users as compared with never-users. After adjusting for age, body mass index (BMI), number of chronic health disorders, fracture history, and time since menopause (independent risk factors) the corresponding risks were 0.67 (0.55-0.81) and 0.53 (0.37-0.74), respectively. The respective adjusted risks for continuous HRT users were 0.62 (0.48-0.79) and 0.41 (0.26-0.67). The adjusted risk of other than distal forearm fracture was 0.74 (0.55-0.98). The results suggest that HRT has a beneficial effect on prevention of fractures in general and on that of distal forearm fracture in particular in early postmenopausal women. PMID- 11874245 TI - Reductions in bone mass, structure, and strength in axial and appendicular skeletons associated with increased turnover after ovariectomy in mature cynomolgus monkeys and preventive effects of clodronate. AB - Over 16 months, we evaluated the effects of ovariectomy (OVX) and bisphosphonate clodronate (CLO) on bone in 48 cynomolgus monkeys (9-15 years old) fed a normal calcium diet. We established three OVX groups (oral CLO at 0 [OVX control], 12, or 60 mg/kg per day) and one sham-operated (SHAM) group. At 16 months, the bone mineral density (BMD) values (percentage of group baseline; OVX control vs. SHAM) for lumbar bone (L3-L5), proximal femur, midfemur, radius, and tibia were -2.6% versus 11.2%, -3.5% versus 8.9%, -3.0% versus 9.0%, -5.5% versus 15.7%, and -6.7% versus 13.9%, respectively. In OVX control (i) tibia showed significant loss of bone mineral content (BMC; vs. baseline), (ii) urinary deoxypyridinoline (DPD) and serum osteocalcin (OC) levels increased (peak = 182% and 168%, respectively, of SHAM), (iii) in lumbar bone and midfemur, ultimate load (UL) was reduced (vs. SHAM), (iv) in lumbar bone, trabecular bone-formation rates (BFRs) were not changed significantly, but tibial endocortical and intracortical bone formation rates were significantly raised (vs. SHAM), (v) the volumetric BMD (vBMD) and geometry of the tibial cortex (measured by peripheral quantitative computed tomography [pQCT]) were significantly reduced (vs. SHAM). CLO, 60 mg/kg per day but not 12 mg/kg per day, significantly inhibited OVX-induced changes, age dependent increases in bone mass, and ability to maintain structure. Thus, in OVX mature cynomolgus monkeys (possibly, a unique model of the cortical bone loss secondary to estrogen deficiency), the post-OVX increases in systemic bone markers were slight, but stimulation of local turnover in the cortical envelope was enough to cause bone loss (more so in tibia than in lumbar trabecular bone). High-dose CLO prevented these changes. PMID- 11874246 TI - Associations of calcium intake and physical activity with bone density and size in premenopausal and postmenopausal women: a peripheral quantitative computed tomography study. AB - The purpose of this cross-sectional study was to examine the impact of long-term physical activity (PA) and calcium intake on non-weight-bearing radius and weight bearing tibia. Altogether, 218 healthy, nonsmoking women, [92 premenopausal women, mean age, 32.6 years (SD, 2.2 years), and 126 postmenopausal women, mean age, 67.3 years (SD, 2.0 years)] participated. The subjects were divided according to their habitual levels of physical activity (PA+ or PA-) and calcium intake (Ca+ or Ca-). The distal end and shaft regions of the radius and tibia were evaluated with peripheral quantitative tomography (pQCT). For the shaft regions, bone mineral content (BMC), cortical cross-sectional area (CoA), cortical density (CoD), and bone strength index, that is, 1-11.9% of the density weighted section modulus (BSI) were determined. For the distal ends, BMC, total cross-sectional area (ToA), trabecular density (TrD), and BSI were determined. The BMC at the distal radius in the young PA+ group was 6.6% (95% CI, 1- to 11.9%) lower than that of the PA- group. A similar nonsignificant trend was found for the radial shaft. The radial shaft showed a mechanically more competent structure among the older subjects with a BSI 8.5% (95% CI, 1.8-15.6%) higher in the older PA+ group than in the older PA- group. The associations between calcium intake and the radial bone characteristics were systematically positive in both age groups. PA seemed to benefit the distal tibia. In the younger age group the TrD was 6.9% (95% CI, 1.8-12.4%) higher in the PA+ group, and in the elderly the BMC was 5% (95% CI, 0.3-9.9%) higher in the PA+ group than in the PA- group. Note that in the younger age group the ToA was 5.1% (95% CI, 0-9.1%) smaller in the PA+ group than in the PA- group, and in the older age group the ToA was 4.2% (95% CI, -0.3-8.9%) greater in the PA+ group than in the PA- group. The association of PA and bone characteristics at the tibial shaft was positive in both age groups (statistically significant for the older subjects). The tibial shaft BSI of the older PA+ group was 8.6% (95% CI, 2.6-14.9%) better than that of the old PA- group. There was no association between calcium intake and the tibial bone characteristics in either age group. In conclusion, high calcium intake was positively associated with a mechanically competent structure in the radius among both younger and older women, whereas the influence of PA did not become apparent until older ages. PA seemed to benefit particularly the weight-bearing tibia, whereas calcium intake was not associated with the tibia. PMID- 11874247 TI - Decreased expression of MAP-2 and GAD in the brain of cats infected with feline immunodeficiency virus. AB - HIV-1 infection is often complicated by the dysfunction of central nervous system (CNS). Degenerative neuronal changes as well as neuronal loss have been documented in individuals with acquired immunodeficiency syndrome. Feline immunodeficiency virus (FIV) causes similar CNS manifestation and FIV infected cats provide an animal model for human immunodeficiency virus infection in humans. In this study, we examined the brain of FIV-infected cats and controls with immunohistochemical techniques using antibodies to microtubule-associated protein 2 (MAP-2) and glutamic acid decarboxylase (GAD). We found a significant decrease in expression of MAP-2 and GAD in neurons of infected animals compared to controls. In contrast, the expression of neurofilaments and glial fibrillary acidic protein was rather increased. The changes observed in the brain were similar to those seen in humans undergoing the normal aging process as well as those suffering from neurological diseases like Alzheimer's disease and other dementing disorders. These changes in the feline brain give insight into the deleterious effects of FIV on the CNS. PMID- 11874248 TI - Activation of mitochondrial ATP-dependent protease by peptides and proteins. AB - We examined the effect of peptides or protein on the proteolytic and ATPase activities of mitochondrial ATP-dependent LON protease purified from bovine adrenal cortex. Peptides/proteins including angiotensin I which stimulated ATPase activity without hydrolysis of any peptide bonds stimulated proteolysis of 125I labeled substrates at low concentrations; whereas at high concentrations they competitively inhibited proteolysis, thus displaying a biphasic activity profile. All peptides and proteins thus examined stimulated degradation of 125I-labeled substrates. When an ATP analog was substituted for ATP, only inhibition; i.e., no stimulation, of proteolysis by unlabeled peptides was observed. Without activator peptides, degradation of [125I] peptides was higher in the presence of an ATP analog than that in the presence of ATP. ADP, a product of the ATPase reaction, inhibited the proteolytic activity in the absence of an activator peptide but not in its presence. From analogy to E. coli ATP-dependent protease La (LON), we suggest that the activator peptides stimulated the proteolysis by releasing enzyme-bound ADP. PMID- 11874249 TI - Concentrations of trace elements in sweat during sauna bathing. AB - Trace elements in sweat during sauna bathing were assessed. Sweat collected by the whole body method was compared with that collected by the arm bag method. The sweat samples were collected from ten healthy male adults aged 22-26 years, by heat exposure in dry sauna bathing (60 degrees C, 30 minutes). Concentrations of major (Na, Cl, K, Ca, P and Mg) and trace (Zn, Cu, Fe, Ni, Cr and Mn) elements in sweat tended to be lower in the arm bag method than in the whole body method. It was found that Ca, Mg, Fe and Mn concentrations in the arm bag method were significantly lower than those in the whole body method. The amount of trace elements in sweat measured by the arm bag method was less than that by the whole body method; significant differences were observed in Fe and Mn amounts. These observations suggest that excretion of trace elements by sweating induces trace element decrease. Therefore, athletes and workers who work in a hot environment and sweat much habitually should ingest adequate amounts of trace elements. PMID- 11874250 TI - Effects of dobutamine on coronary flow velocity response and their relations to age. AB - The purpose of this study was to assess the effects of low-dose dobutamine on left ventricular (LV) functional and coronary flow reserves using transthracic echocardiography. The study group consisted of 30 children aged from 5 months to 16 years (mean 4.8 +/- 4.4 years). Echocardiographic studies were repeated before and during dobutamine infusion (5 microg/kg per minute). The peak diastolic velocity in the left descending coronary artery (LAD) was recorded by pulsed Doppler under the guidance of color Doppler flow mapping. The coronary flow velocity (CFV) response was calculated as the ratio of LAD peak flow velocity at dobutamine infusion to basal LAD peak flow velocity. Left ventricular contractility was calculated by two-dimensionally directed M-mode echocardiography. The rate-corrected mean velocity of circumferential fiber shortening (mVcfc) and LV end-systolic wall stress (ESS) were used as indices of contractility. Adequate spectral Doppler recordings of the LAD peak flow velocity for the assessment of CFV response were obtained in 26 of 30 patients (87%). The LAD peak flow velocity at dobutamine infusion increased significantly compared with the basal values. The CFV response in the younger children was low and increased significantly with age. The CFV response did not show significant correlations with the changes in heart rate, systolic blood pressure, rate pressure product, nor ESS during dobutamine infusion. However, a significant relationship between the CFV response and the percent change of mVcfc was observed. In the present study using high frequency transthoracic echocardiography, we demonstrated the age-related changes in CFV response and LV functional reserve by dobutamine infusion. Responses of LV contractility and coronary flow to dobutamine are less sensitive in youngerchildren and increased with increasing age. PMID- 11874251 TI - Clinical investigation of the lesions responsible for sensory disturbance in Minamata disease. AB - To clarify the lesions responsible for sensory disturbance in Minamata disease (MD), we clinically investigated the characteristics of sensory disturbance. In all patients with the classical type MD, two-point discrimination was severely disturbed, but the involvement of superficial sensation was relatively mild. On short-latency somatosensory evoked potential study, the component corresponding to N20 was completely absent with normal N9, N11, and N13 components. Although 14 of 38 chronic MD patients demonstrated intact superficial sensation, 10 of these 14 showed mild to moderate disturbance in two-point discrimination. The two-point discrimination in chronic MD patients was significantly high irrespective of the disturbance of superficial sensation. These findings suggest that the sensory disturbance of MD patients may mainly be caused by a lesion in the sensory cortex rather than in the peripheral nerves. However, other foci could be also responsible for the sensory impairment, since 9 of 38 chronic MD patients showed intact two-point discrimination. PMID- 11874252 TI - A case of hemosuccus pancreaticus associated with hereditary pancreatitis. AB - We report a 25-year-old male with hemosuccus pancreaticus associated with hereditary pancreatitis. He was originally diagnosed as having familial chronic pancreatitis at the age of 12, because his brother was also diagnosed as having pancreatitis. No history of pancreatitis was found in their parents. The patient was admitted because of a growing pancreatic pseudocyst. While he had undergone conservative treatment for the pseudocyst, computed tomography incidentally revealed a pancreatic pseudoaneurysm. Endoscopic examination revealed spontaneous bleeding from the major papilla. Interventional embolization was successfully performed. An R122H mutation in the cationic trypsinogen gene was identified in this patient, his brother, and his mother, indicating that they have hereditary pancreatitis. To our knowledge, this is the first report of hemosuccus pancreaticus associated with hereditary pancreatitis. Mutational screening is useful for the diagnosis of hereditary pancreatitis, especially in patients whose diagnosis is inconclusive based on the traditional clinical criteria. PMID- 11874253 TI - The paradox of health care. AB - The term "paradox" signifies a contradiction of some sort. Modern health care appears to be rich in contradictions, and it is claimed to be paradoxical in a number of ways. In particular health care is held to be a paradox itself: it is supposed to do good, but is accused of doing harm. The objective of this article is to investigate whether the concept of paradox can serve as a framework for analysing pressing problems in modern health care. To pursue this, three distinctive levels of paradox are identified: resolvable paradoxes, antinomies and aporias. The analysis reveals that when facing the challenges of modern health care the focus of attention should be to resolve the resolvable paradoxes, to acknowledge the antinomies and to learn to live with the aporias. PMID- 11874254 TI - Foetal images: the power of visual technology in antenatal care and the implications for women's reproductive freedom. AB - Continuing medico-technical progress has led to an increasing medicalisation of pregnancy and childbirth. One of the most common technologies in this context is ultrasound. Based on some identified 'pro-technology feminist theories', notably the postmodernist feminist discourse, the technology of ultrasound is analysed focusing mainly on social and political rather than clinical issues. As empirical research suggests, ultrasound is welcomed by the majority of women. The analysis, however, shows that attitudes and decisions of women are influenced by broader social aspects. Furthermore, it demonstrates how the visual technology of ultrasound, in addition to other reproductive technology in maternity care, is linked to the 'personification' of the foetus and has therefore contributed to a new image of the foetus. The exploration of these issues challenges some arguments of feminist discourse. It draws attention to possible adverse implications of the technology for women's reproductive freedom and indicates the importance of the topic for political discussions. PMID- 11874255 TI - 'Transgressing venues': 'health' studies, cultural studies and the media. AB - This paper looks at how the strategies of media and cultural studies can be applied to the health studies field. This relationship, however, has been met with resistance due to a number of status debates. We argue the importance of fostering links between these 'disciplines' namely because the definition of what constitutes 'health' has been broadened and is inscribed in most forms of popular media. Using the example of the 'health and lifestyle' debate, we argue that the media informs cultural understandings about requirements for living and is therefore a crucial area of analysis for health studies (and cultural/media studies) students. We are also concerned with analysing how the media contributes to the construction and regulation of 'healthy' subjects making links with public health discourses and current debates in the field of media and health. PMID- 11874256 TI - NHS trust chief executives as heroes? AB - This paper presents a reading of the transcripts of interviews with NHS Trust Chief Executives. Using a poststructuralist understanding of the interviews, it privileges a reading that (ironically) represents these Chief Executives as heroes. Following the classic hero story line, they leave the civilized order of home and journey into a threatening wilderness where they encounter dangerous and magical things but overcome them all because of their masculine characteristics such as rationality, strength and resourcefulness. One way in which these stories can be understood to have significance is that they (misleadingly but powerfully) portray management as obvious and necessary by evocatively drawing on a myth of ancient origin. The piece concludes with some reflections on the ontological implications of the analysis and reflexive comments on the production of truth as a problem. PMID- 11874257 TI - Is subjective well-being a useful parameter for allocating resources among public interventions? AB - Scarce public resources require trade-offs between competing programs in different sectors, and the careful allocation of fixed resources within a single sector. This paper argues that a general quality of life instrument encompassing health-related and non-health-related components is suitable for determining the best trade-offs between sectors. Further, this paper suggests that subjective well-being shows the properties crucial to a general quality of life measure and has additional advantages that makes it particularly useful for the allocation of public and health care resources. The paper argues that Western societies are in an unusually prosperous situation today which allows to concentrate efforts not only on reducing harm but also on improving positive states of health. Further, subjective well-being can be evaluated from the patient's perspective and incorporates a valuation of life expectancy. Criteria required for an appropriate questionnaire that measures subjective well-being are presented. PMID- 11874258 TI - Power issues in the doctor-patient relationship. AB - Power is an inescapable aspect of all social relationships, and inherently is neither good nor evil. Doctors need power to fulfil their professional obligations to multiple constituencies including patients, the community and themselves. Patients need power to formulate their values, articulate and achieve health needs, and fulfil their responsibilities. However, both parties can use or misuse power. The ethical effectiveness of a health system is maximised by empowering doctors and patients to develop 'adult-adult' rather than 'adult child' relationships that respect and enable autonomy, accountability, fidelity and humanity. Even in adult-adult relationships, conflicts and complexities arise. Lack of concordance between doctors and patients can encourage paternalism but may be best resolved through negotiated care. A further area of conflict involves the 'double agency' of doctors for both patients and the community. Empowerment of all players is not always possible but is most likely where each party considers and acknowledges power issues. PMID- 11874259 TI - Bioethics at the crossroad. AB - Bioethics and its offspring Health-care Ethics have a variety of uses and obligations among which and perhaps most importantly is their social obligation. This paper raises questions as to Bioethics fulfilling the necessary criteria for a profession, suggests that it can serve as a link between individual and communal problems, discusses the task of health-care ethics as well as ways of teaching it, lists some of the obligations of health-care ethics professionals and discusses the dangers to and failings of these health-care professionals today. It concludes that we are at a crossroads in which we must choose between our own personal security and comfort and fulfilling our social role. PMID- 11874260 TI - Management of hepatocellular carcinoma. AB - This article reviews the recent advances in the management of hepatocellular carcinoma (HCC). Partial hepatectomy still remains the mainstay of therapy for early HCC. The limits of liver resection have been extended with the use of a multi-modality approach, and neoadjuvant/adjuvant treatment has improved the results of liver resection. Orthotopic liver transplantation works better than partial hepatectomy in a subgroup of patients with poor liver function and with early HCC. For locally advanced HCC, which is still confined to the liver, debulking surgery gives excellent palliation and prolongs patient survival. Non operative local ablative therapy shows encouraging results. Hepatic artery chemoembolisation has been shown, by systemic review or meta-analysis, to have no significant impact on patient survival. Initial results with transarterial radioembolisation are promising. For patients with advanced HCC, systemic chemotherapy has little clinical benefit. The results of systemic immunotherapy or systemic tamoxifen on HCC are controversial and need further evaluation. Early results of chemoimmunotherapy are encouraging. PMID- 11874261 TI - Restorative proctocolectomy: one surgeon's experience of a now routine procedure in a district general hospital. AB - Restorative proctocolectomy is now the operation of choice for the definitive management of ulcerative colitis and familial adenomatous polyposis coli. The aim of this review is to evaluate clinical and functional results of the first fifty patients treated with restorative proctocolectomy over a period of seven years. A total of 50 patients (28 males, 22 females) underwent restorative proctocolectomy over a period of seven years in our district general hospital. A retrospective analysis of hospital notes was undertaken and supplemented with a patient questionnaire. Forty-three patients had ulcerative colitis and seven had familial adenomatous polyposis coli. Two pouch designs were used, a four limbed "pear pouch" (94%) and a two limb "J" pouch (6%). The majority (98%) of patients had a stapled ileal pouch-anal anastomosis and a similar number had a defunctioning ileostomy. There was no mortality. Early complications (within 30 days of operation) occurred in 18(36%) patients. Late complications were seen in 22 (44%) patients. The median number of daily bowel movements was four (range 1-10). Pouchitis was seen in ten patients (20%). Restorative proctocolectomy is a safe procedure for patients with ulcerative colitis and familial adenomatous polyposis coli. Although total morbidity is appreciable, functional results are generally good and patient satisfaction is high. PMID- 11874262 TI - Ligation of the splenic artery in operations of the spleen in children. AB - BACKGROUND: Every effort is made during surgery of the spleen to preserve splenic function. This serves the purpose of maintaining the integrity of the defence mechanisms of the patient against infection in the short - and long-term. With ligation of the splenic artery prior to excision of splenic lesions and splenic repair we aimed to secure control of peri- operative bleeding, thus, making feasible the preservation of the spleen. METHODS: Six patients with splenic pathology aged 5-14 years old (four boys and two girls) were operated on for splenic pathology between 1994-1999. Three had splenic cysts (dermoid), one had a hamartoma and 2 had hydatid cysts. Investigations included radiography, ultrasound scan, computerised tomography and scintigraphy. All patients were administered antibiotics and pneumovax pre-operatively. Ligation of the splenic artery was carried out with ligaclips. Congenital anomalies or hydatid cysts were excised and repair of the spleen with splenorrhaphy was performed. Peri-operative blood loss was minimal and this aided greatly in performing the excision of the splenic lesions and the repair of the spleen. RESULTS: All patients had an uncomplicated post-operative course. Scintigraphy in the follow-up from 6 months after the operation showed good splenic function. Laboratory investigations did not show decreased levels of immunological factors in the early or later post- operative period. CONCLUSION: Splenic artery ligation reduced the amount of bleeding and allowed for the preservation of the spleen in all patients in this study. PMID- 11874263 TI - Haematuria investigation based on a standard protocol: emphasis on the diagnosis of urological malignancy. AB - OBJECTIVE: To audit the findings of a standard investigation protocol for haematuria with emphasis on the diagnosis of urological malignancy. METHODS: Data were prospectively collected on haematuria referrals to one centre over a 5 year period. The standard protocol of investigation included flexible cystoscopy, urine cytology and culture, upper tract imaging, consisting of a renal tract ultrasound scan and a radiograph of kidney-ureter-bladder (KUB), proceeding to an intravenous urogram (IVU) in selected patients. RESULTS: 1046 patients were examined; 63% (n = 657) had microscopic haematuria and 37% (n = 389) had frank haematuria. No malignancy was found in patients with microscopic haematuria below 50 years of age. The findings of malignancy were not associated with either the sex or duration of symptoms in either groups. No association between the presence of symptoms and the finding of malignancy was observed in the microscopic haematuria group. Twenty five percent of patients presenting with frank haematuria had malignancy compared with 3.7% of patients with microscopic haematuria (p < 0.0001). The type of haematuria (frank or microscopic) was not predictive of grade or stage of malignancy. Of patients under 70 years with frank haematuria, males were more likely than females to have malignancy. This higher risk was not observed in older patients. Urine cytology had a poor predictive value for detection of malignancy with a sensitivity of only 25%. CONCLUSION: Full investigation of all patients with frank haematuria and those with microscopic haematuria above 50 years of age, is well justified. Patients under 50 years with microscopic haematuria should have a lower priority for investigation. PMID- 11874264 TI - Endoscopic anterior cruciate ligament reconstruction using a four strand hamstring tendon construct. AB - Disruption of the anterior cruciate ligament (ACL) may result in recurrent episodes of giving way of the knee with the risk of concomitant damage to the menisci and chondral surfaces. Surgical reconstruction for ACL ligament deficiency is aimed at restoring normal knee kinematics, thereby, allowing for return to pre-injury function. Endoscopic reconstruction of the ACL using a four strand tendon autograft is a well documented, prospectively evaluated methodology. This article outlines the authors' technique and identifies key points of the surgical procedure. PMID- 11874265 TI - Atraumatic rupture of the spleen in adults. AB - Atraumatic rupture of the spleen is an uncommon but important clinical entity. Unfortunately, the diagnosis is often missed due to the absence of any history of trauma. Various aspects of atraumatic rupture of the spleen, including those of 'pathologic' and 'spontaneous' rupture of the spleen, have been reviewed. Increased awareness of this condition would enhance early diagnosis and effective treatment. Further research is required to identify the possible risk factors associated with spontaneous rupture of the spleen. PMID- 11874266 TI - Review: platelet glycoprotein IIb/IIIa blockers for PCI or acute coronary syndromes reduce death and MI but increase bleeding. PMID- 11874267 TI - Reteplase plus abciximab and reteplase alone led to similar 30-day mortality rates in acute MI. PMID- 11874268 TI - Tenecteplase plus enoxaparin or abciximab was better than tenecteplase plus unfractionated heparin for acute MI. PMID- 11874269 TI - Abciximab did not reduce death or myocardial infarction in the acute coronary syndrome without early revascularization. PMID- 11874270 TI - Clopidogrel plus aspirin was effective but increased bleeding in acute coronary syndromes without ST-segment elevation. PMID- 11874271 TI - A nurse-led intervention reduced risk factors, anxiety, and depression in patients waiting for CABG. PMID- 11874272 TI - Revascularization led to less angina and fewer adverse cardiac events than did optimal medical care in angina pectoris in the elderly. PMID- 11874273 TI - The Dietary Approaches to Stop Hypertension diet lowered systolic blood pressure in stage-1 isolated systolic hypertension. PMID- 11874274 TI - Continuous and intermittent sibutramine were equally effective at 44 weeks for reducing weight in obese persons. PMID- 11874275 TI - Review: thrombolysis increases short-term death and intracranial hemorrhage but decreases long-term death or dependence. PMID- 11874276 TI - Combined therapy with indapamide and perindopril but not perindopril alone reduced the risk for recurrent stroke. PMID- 11874277 TI - Levodopa plus carbidopa before physiotherapy increased motor recovery after stroke. PMID- 11874278 TI - Review: studies on the cardiovascular effects of selective COX-2 inhibitors show mixed results. PMID- 11874279 TI - Review: mucolytic drugs reduce exacerbations, illness days, and antibiotic use in chronic bronchitis and COPD. PMID- 11874280 TI - Prone positioning for acute respiratory failure improved short-term oxygenation but not survival. PMID- 11874281 TI - Single-dose doxycycline prevented Lyme disease after an Ixodes scapularis tick bite. PMID- 11874282 TI - Prolonged antibiotic treatment did not relieve chronic symptoms in Lyme disease. PMID- 11874283 TI - Itraconazole was as effective as amphotericin B for fever and neutropenia in cancer and led to fewer adverse events. PMID- 11874284 TI - Donepezil improved the clinical state and quality of life in moderate-to-severe Alzheimer disease. PMID- 11874285 TI - A community support program for dementia delayed but did not reduce institutionalization over 2 years. PMID- 11874286 TI - Review: behavioral interventions show the most promise for the chronic fatigue syndrome. PMID- 11874287 TI - Review: antidepressants improve headache in patients with chronic headache. PMID- 11874288 TI - Review: long-term lithium treatment lowers suicide risk in major affective disorder. PMID- 11874289 TI - Review: problem-solving treatment after deliberate self-harm improves depression, hopelessness, and personal problems. PMID- 11874290 TI - Physiotherapy, aerobics, and training devices reduced pain intensity and frequency in chronic low-back pain. PMID- 11874291 TI - Zinc or vitamin A reduced diarrhea in young, poor Bangladeshi children. PMID- 11874292 TI - A rice-based diet with green banana or pectin reduced diarrhea in infants better than a rice-alone diet. PMID- 11874293 TI - B-natriuretic peptide levels had high sensitivity and specificity for detecting abnormal ventricular function. PMID- 11874294 TI - Review: CT angiography and magnetic resonance imaging are the best less-invasive tests for renal artery stenosis. PMID- 11874295 TI - Clinical and radiologic evaluation had moderate sensitivity and specificity for detecting idiopathic pulmonary fibrosis. PMID- 11874296 TI - Children infected with meningitis before 1 year of age were at increased risk for disability at 5 years. PMID- 11874297 TI - Low-level prenatal alcohol exposure was associated with adverse behavioral outcomes in children at 6 to 7 years of age. PMID- 11874298 TI - Review: multidisciplinary CHD management programs improve the process of care and reduce hospitalizations. PMID- 11874299 TI - Rule-based computerized reminders increased ordering of preventive services in an inpatient setting. PMID- 11874300 TI - An intranasal influenza vaccine for the prevention of influenza in healthy children was cost-effective. PMID- 11874301 TI - The TIMI risk score predicted mortality in patients with ST-elevation myocardial infarction. PMID- 11874302 TI - A standardized sequential clinical examination identified probable causes of syncope in 69% of patients. PMID- 11874303 TI - Clinical expertise in the era of evidence-based medicine and patient choice. PMID- 11874305 TI - Trends in the incidence and survival of patients with hospitalized myocardial infarction, Olmsted County, Minnesota, 1979 to 1994. AB - BACKGROUND: Evidence indicates that deaths from coronary disease are decreasing less in elderly persons and women. Understanding the determinants of these trends is important for prevention. OBJECTIVE: To test the hypothesis that trends in incidence and survival of hospitalized myocardial infarction differ according to sex and age. DESIGN: Longitudinal observational study. SETTING: Community dwelling persons in Olmsted County, Minnesota. PATIENTS: 5117 patients who had an estimated 1820 incident myocardial infarctions from 1979 to 1994. MEASUREMENTS: Myocardial infarctions were validated by using epidemiologic criteria. Rates were directly adjusted to the age distribution of the 2000 U.S. population. RESULTS: Of the 1820 incident infarctions, 44% occurred in women and 36% in persons 75 years of age or older. In 1979, the age-adjusted incidence of myocardial infarction was 205 per 100 000 persons (95% CI, 162 to 247 per 100 000 persons). Between 1979 and 1994, the age-adjusted incidence of myocardial infarction decreased by 8% (CI, -23% to 10%) in men but increased by 36% (CI, 9% to 70%) in women. A 31% decrease in the incidence of infarction over time was observed in men 40 years of age compared with a 49% increase in women 80 years of age. Survival improved predominantly in younger persons. CONCLUSIONS: Over time, the incidence of hospitalized infarction decreased in men but increased in women and elderly persons. Survival benefits were clustered among younger persons. These results suggest that both incidence and survival contribute to the contrasting mortality trends by age and sex and that the burden of coronary disease has shifted toward elderly persons, a finding that has public health implications in an aging population. PMID- 11874307 TI - Survival associated with 5-fluorouracil-based adjuvant chemotherapy among elderly patients with node-positive colon cancer. AB - BACKGROUND: Randomized clinical trials have demonstrated the efficacy of adjuvant 5-fluorouracil (5-FU)-based chemotherapy after surgical resection of node positive colon cancer. Although this treatment became the standard in 1990 following a National Institutes of Health Consensus Conference, among those at least 65 years of age it is less likely to be offered to older or nonwhite patients. OBJECTIVE: To determine the association between 5-fu-based chemotherapy and survival in older patients. DESIGN: Retrospective cohort study. SETTING: Combined database of the National Cancer Institute's Surveillance, Epidemiology, and End Results (SEER) program and Medicare. PATIENTS: 4768 patients 65 years of age or older who received a diagnosis of node-positive colon cancer from 1992 to 1996, were covered by Medicare Parts A and B, and resided in the population covered by the SEER program. MEASUREMENTS: Propensity scores to control for known predictors of receiving treatment, Cox proportional hazards models to assess the association of 5-FU therapy with survival, and sensitivity analyses to estimate the possible effects of unknown confounders. RESULTS: Fifty-two percent of patients received 5-FU therapy. For this sample, the hazard ratio for death associated with 5-FU therapy was 0.66 (95% CI, 0.60 to 0.73). Confounding could have accounted for this association only if an unmeasured confounder were extremely unequally distributed between the treated and untreated groups or increased mortality by at least 50%. CONCLUSIONS: 5-Fluorouracil adjuvant therapy is significantly associated with reduced mortality in older patients, similar to the association found in randomized, controlled trials among younger patients. More frequent use of 5-FU therapy in older patients would probably reduce death from colon cancer. PMID- 11874308 TI - Burnout and self-reported patient care in an internal medicine residency program. AB - BACKGROUND: Burnout is a syndrome of depersonalization, emotional exhaustion, and a sense of low personal accomplishment. Little is known about burnout in residents or its relationship to patient care. OBJECTIVE: To determine the prevalence of burnout in medical residents and explore its relationship to self reported patient care practices. DESIGN: Cross-sectional study using an anonymous, mailed survey. SETTING: University-based residency program in Seattle, Washington. PARTICIPANTS: 115 internal medicine residents. MEASUREMENTS: Burnout was measured by using the Maslach Burnout Inventory and was defined as scores in the high range for medical professionals on the depersonalization or emotional exhaustion subscales. Five questions developed for this study assessed self reported patient care practices that suggested suboptimal care (for example, "I did not fully discuss treatment options or answer a patient's questions" or "I made...errors that were not due to a lack of knowledge or inexperience"). Depression and at-risk alcohol use were assessed by using validated screening questionnaires. RESULTS: Of 115 (76%) responding residents, 87 (76%) met the criteria for burnout. Compared with non-burned-out residents, burned-out residents were significantly more likely to self-report providing at least one type of suboptimal patient care at least monthly (53% vs. 21%; P = 0.004). In multivariate analyses, burnout--but not sex, depression, or at-risk alcohol use- was strongly associated with self-report of one or more suboptimal patient care practices at least monthly (odds ratio, 8.3 [95% CI, 2.6 to 26.5]). When each domain of burnout was evaluated separately, only a high score for depersonalization was associated with self-reported suboptimal patient care practices (in a dose-response relationship). CONCLUSION: Burnout was common among resident physicians and was associated with self-reported suboptimal patient care practices. PMID- 11874309 TI - Paying research subjects: an analysis of current policies. AB - BACKGROUND: Few data are available on guidelines used by research organizations to make decisions about paying subjects. OBJECTIVE: To analyze existing guidance regarding payment of research subjects and to identify common characteristics and areas for further research. DESIGN: Descriptive content analysis of policies. MEASUREMENTS: Written policies and rules of thumb about paying subjects from 32 U.S. research organizations. RESULTS: Of 32 organizations, 37.5% had written guidelines about paying subjects; all but 1 reported having rules of thumb. Few (18.8%) were able to provide a confident estimate of the proportion of studies that pay subjects. Organizations reported that investigators and institutional review boards make payment decisions and that both healthy and ill subjects in some studies are paid for their time (87%), for inconvenience (84%), for travel (68%), as incentive (58%), or for incurring risk (32%). Most organizations require that payment be prorated (84%) and described in the consent document (94%). CONCLUSIONS: Most organizations pay some research subjects, but few have written policies on payment. Because investigators and institutional review boards make payment decisions with little specific guidance, standards vary. PMID- 11874310 TI - Acupuncture: theory, efficacy, and practice. AB - Traditionally, acupuncture is embedded in naturalistic theories that are compatible with Confucianism and Taoism. Such ideas as yin-yang, qi, dampness, and wind represent East Asian conceptual frameworks that emphasize the reliability of ordinary, human sensory awareness. Many physicians who practice acupuncture reject such prescientific notions. Numerous randomized, controlled trials and more than 25 systematic reviews and meta-analyses have evaluated the clinical efficacy of acupuncture. Evidence from these trials indicates that acupuncture is effective for emesis developing after surgery or chemotherapy in adults and for nausea associated with pregnancy. Good evidence exists that acupuncture is also effective for relieving dental pain. For such conditions as chronic pain, back pain, and headache, the data are equivocal or contradictory. Clinical research on acupuncture poses unique methodologic challenges. Properly performed acupuncture seems to be a safe procedure. Basic-science research provides evidence that begins to offer plausible mechanisms for the presumed physiologic effects of acupuncture. Multiple research approaches have shown that acupuncture activates endogenous opioid mechanisms. Recent data, obtained by using functional magnetic resonance imaging, suggest that acupuncture has regionally specific, quantifiable effects on relevant brain structures. Acupuncture may stimulate gene expression of neuropeptides. The training and provision of acupuncture care in the United States are rapidly expanding. PMID- 11874311 TI - Stress in medical residency: status quo after a decade of reform? AB - BACKGROUND: Although curricular reforms have attempted to address sources of stress in medical residency, no recent studies have examined the financial or emotional situations of current medicine residents. OBJECTIVE: To question medicine residents about financial status, educational debt, moonlighting, and psychological issues. DESIGN: Survey distributed in a nonrandomized fashion to medicine residents. SETTING: All 415 U.S. medicine residency programs. RESULTS: According to the questionnaire responses submitted by the 4128 (18%) participating residents, a substantial number of residents had financial and emotional distress that could have interfered with training. The reported educational debt was at least $50 000 for 1657 (42%) of the respondents and at least $100 000 for 737 (19%). The monthly disposable income was $100 or less for 1620 (43%) of the residents, and 637 residents (16%) could not afford safe housing. Among respondents in their 2nd through 5th year of postgraduate training, 2187 (52%) had insufficient funds to purchase books and equipment, and 678 (29%) could not afford the required fees for the American Board of Internal Medicine certifying examination; 2659 (33%) worked as moonlighters, and this percentage increased progressively with increasing educational debt. Four or five depressive symptoms during residency were reported by 1461 (35%) residents. Eight hundred ninety-nine residents (23%) thought they had become less humanistic over the course of their residency training; 2347 (61%) reported becoming more cynical. Female residents were more likely than male residents to report increased cynicism and multiple depressive symptoms. Increased cynicism and depressive symptoms were associated with increasing educational debt. CONCLUSIONS: Despite recent curricular reforms, an alarming number of current medicine residents report depressive symptoms, increasing cynicism, and decreasing humanism, which were associated with increasing educational debt and a need to moonlight for financial survival. Ongoing curricular reform, legislative relief from early loan repayment, and salary increases may be necessary to address these problems. PMID- 11874313 TI - Heeding the plea to deal with resident stress. PMID- 11874312 TI - Who is sicker: patients--or residents? Residents' distress and the care of patients. PMID- 11874314 TI - Physician-industry relations. Part 1: individual physicians. AB - This is part 1 of a 2-part paper on ethics and physician-industry relationships. Part 1 offers advice to individual physicians; part 2 gives recommendations to medical education providers and medical professional societies. Physicians and industry have a shared interest in advancing medical knowledge. Nonetheless, the primary ethic of the physician is to promote the patient's best interests, while the primary ethic of industry is to promote profitability. Although partnerships between physicians and industry can result in impressive medical advances, they also create opportunities for bias and can result in unfavorable public perceptions. Many physicians and physicians-in-training think they are impervious to commercial influence. However, recent studies show that accepting industry hospitality and gifts, even drug samples, can compromise judgment about medical information and subsequent decisions about patient care. It is up to the physician to judge whether a gift is acceptable. A very general guideline is that it is ethical to accept modest gifts that advance medical practice. It is clearly unethical to accept gifts or services that obligate the physician to reciprocate. Conflicts of interest can arise from other financial ties between physicians and industry, whether to outside companies or self-owned businesses. Such ties include honorariums for speaking or writing about a company's product, payment for participating in clinic-based research, and referrals to medical resources. All of these relationships have the potential to influence a physician's attitudes and practices. This paper explores the ethical quandaries involved and offers guidelines for ethical business relationships. PMID- 11874315 TI - Physician-industry relations. Part 2: organizational issues. AB - This is part 2 of a 2-part paper on ethics and physician-industry relationships. Part 1 offers advice to individual physicians; part 2 gives recommendations to medical education providers and medical professional societies. Industry often sponsors programs for graduate and continuing medical education, as well as major events of medical professional societies. Industry is an abundant source of advances in medicine and technology and plays a crucial role in disseminating up to-date medical information. Although industry information fills an important need, studies suggest that it is often biased. Providers of graduate and continuing medical education have a duty to present objective and balanced information to their participants; thus, they should not accept any funds that are contingent on a sponsor's ability to shape programming. Medical educators need to evaluate and control the planning, content, and delivery of education provided under their auspices. They should disclose industry sponsorship to students, faculty, and continuing medical education participants and should adopt explicit organizational policies about acceptable and unacceptable interactions with industry. Medical professional societies have a duty to promote the independent judgment and professionalism of their members. Organizers of industry sponsored meetings should clearly separate product promotion from impartial medical education. Adopting specific policies for dealing with industry sponsorship can also help professional societies guard against outside influence. The American College of Physicians--American Society of Internal Medicine's core ethical principles for external funding and relationships serve as an example. PMID- 11874316 TI - On being a doctor. Blockers. PMID- 11874318 TI - The crisis in local institutional review boards. PMID- 11874319 TI - The crisis in local institutional review boards. PMID- 11874320 TI - The crisis in local institutional review boards. PMID- 11874322 TI - Should we screen for depression in primary care? PMID- 11874324 TI - Misconceptions about Lyme disease: confusions hiding behind ill-chosen terminology. PMID- 11874327 TI - Summary for patients. Changes in incidence of heart attacks. PMID- 11874328 TI - Summary for patients. Survival of elderly patients with colon cancer treated with chemotherapy. PMID- 11874330 TI - Summary for patients. Paying research subjects: a survey of current policies. PMID- 11874329 TI - Summary for patients. Do medical residents experience burnout? PMID- 11874331 TI - New structural forms in molecular metal phosphonates: novel tri- and hexanuclear zinc(II) cages containing phosphonate and pyrazole ligands. AB - The reaction of ZnCl(2) with tert-butylphosphonic acid and 3,5-dimethylpyrazole in the presence of triethylamine as a hydrogen chloride scavenger affords a trinuclear molecular zinc phosphonate [Zn(3)Cl(2)(3,5-Me(2)Pz)(4)(t-BuPO(3))(2)]. The structure of this compound contains a planar trizinc assembly containing two bicapping mu(3) [t-BuPO(3)](2-) ligands and terminal pyrazole and chloride ligands. In contrast an analogous reaction of ZnCl(2) with phenylphosphonic acid and 3,5-dimethylpyrazole affords a hexanuclear zinc phosphonate [Zn(6)Cl(4)(3,5 Me(2)PzH)(8)(PhPO(3))(4)]. The six zinc centers are arranged in a chairlike conformation. The four phosphonates in this complex also act as bridging tripodal mu(3) [RPO(3)](2-) ligands. PMID- 11874332 TI - New rigid angular dicarboxylic acid for the construction of nanoscopic supramolecules: from a molecular rectangle to a 1-D coordination polymer. AB - A new rigid angular bridging ligand, 7-oxa-dibenzofluorene-3,11-dicarboxylic acid (H(2)L), was synthesized by cyanation of known rac-6,6'-dibromo-1,1'-bi-2 naphthol followed by ring closure and hydrolysis with concentrated sulfuric acid and used for the self-assembly of nanoscopic molecular rectangle [Cu(4)(L)(4)(Py)(8)].2DMF.10H(2)O, 1, and 1-D coordination polymer [Co(2)(L)(2)(Py)(4)].2DMF.2H(2)O, 2. Both 1 and 2 contain open channels occupied by DMF and water guest molecules. Crystal data for 1:[?] triclinic, space group P(-)1, a = 8.869(2) A, b = 16.437(3) A, c = 21.586(4) A, alpha = 78.18(3), beta = 79.19(3), gamma = 83.66(3), U = 3017.0(11) A(3), and Z = 1. Crystal data for 2: triclinic, space group P(-)1, a = 8.254(2) A, b = 12.154(2) A, c = 15.348(3) A, alpha = 95.34(3), beta = 93.38(3), gamma = 94.37(3), U = 1525.1(5) A(3), and Z = 1. PMID- 11874333 TI - A new type of tweezer complex involving a rhenium-rhenium multiple bond that enforces an unusual structure in a dipalladium(II) unit. AB - The substitution of the mu-acetato ligands in cis-Re(2)(mu-O(2)CCH(3))(2)Cl(2)(mu dppm)(2) (1, dppm = Ph(2)PCH(2)PPh(2)) and trans-Re(2)(mu-O(2)CCH(3))(2)Cl(2)(mu dppE)(2) (2, dppE = Ph(2)PC(=CH(2))PPh(2)) by [4-Ph(2)PC(6)H(4)CO(2)](-) occurs with retention of stereochemistry to give cis-Re(2)(mu-O(2)CC(6)H(4)-4 PPh(2))(2)Cl(2)(mu-dppm)(2) (3) and trans-Re(2)(mu-O(2)CC(6)H(4)-4 PPh(2))(2)Cl(2)(mu-dppE)(2) (6), respectively. The uncoordinated phosphine groups in complexes 3 and 6 have been used to form mixed-metal assemblies with Au(I) and Pd(II), including the Re(2)Pd(2) complex cis-Re(2)(mu-O(2)CC(6)H(4)-4 PPh(2))(2)Cl(2)(mu-dppm)(2)(Pd(2)Cl(4)) (5), in which the planar [(P)ClPd(mu Cl)(2)PdCl(P)] unit has the unusual cis structure. The crystal structures of 3 and 5 have been determined. PMID- 11874335 TI - Beyond Bragg scattering: the structure of AgCN determined from total neutron diffraction. AB - Total neutron diffraction has yielded accurate bond lengths, Ag-C = Ag-N = 2.06 A and C-N = 1.16 A, for the disordered crystalline solid AgCN. This information cannot be obtained from analysis of Bragg scattering studies, because the (-Ag-CN )(n) chains are randomly displaced along the chain axis relative to each other by a root-mean-square displacement of 0.24 A at 10 K. These results show the power of total neutron diffraction for determining structure in disordered systems. PMID- 11874334 TI - First [Fe-NO](6) complex with an N(2)S(3)Fe-NO core as a model of NO-inactivated iron-containing nitrile hydratase. Are thiolates and thioethers equivalent donors in low-spin iron complexes? AB - The spectroscopic and structural properties of [(bmmp-TASN)FeNO]BPh(4) (1) (bmmp TASN = 4,7-bis(2'-methyl-2'-mercaptopropyl)-1-thia-4,7-diazacyclononane) have been determined and are compared with the nitric oxide inactivated form of iron containing nitrile hydratase, NHase(dark). [(bmmp-TASN)FeNO]BPh(4) is prepared from the addition of NO to (bmmp-TASN)FeCl followed by addition of sodium tetraphenylborate. [(bmmp-TASN)FeNO]BPh(4) crystallizes from acetonitrile methanol solutions upon ether vapor diffusion as dark blue plates in the monoclinic space group P2(1)/c with a = 11.9521(14) A, b = 11.3238(13) A, c = 26.624(3) A, beta = 98.280(2), and Z = 4. The nu(NO) stretching frequency of 1856 cm(-)(1) and the Mossbauer parameters, delta = 0.06 mm/s and DeltaE(q) = 1.75 mm/s, compare favorably with those of NHase(dark). The similarities of the iron sulfur bond distances to the thiolate, 2.284(2) A and 2.291(2) A, versus thioether, 2.285(2) A, are attributed to the low-spin configuration of the iron. The relationship between this structural observation and the spectroscopic properties of the complex are discussed. PMID- 11874336 TI - Potentiometric investigations of carbonate ion stability in molten cesium and sodium iodides. AB - The reaction of carbonate ion dissociation in molten CsI and NaI was studied in the temperature range from the melting point to 800 (CsI) and 830 degrees C (NaI) by a potentiometric method with the use of a membrane oxygen electrode as an indicator one. The dissociation constant of CO(3)(2-) in the molten iodides was found to increase with the temperature. pK values for CsI are 4.47 (650 degrees C), 4.23 (700 degrees C), 3.89 (750 degrees C), 3.44 (800 degrees C); those for NaI are 4.68 (700 degrees C), 4.39 (750 degrees C), 4.17 (800 degrees C), 3.92 (830 degrees C). The plots of pK vs reverse temperature are linear. Carbonate stability in molten CsI is lower than that in the NaI due to lower stability of Cs(2)CO(3) compared with Na(2)CO(3). PMID- 11874337 TI - High-resolution optical spectroscopy of Na(3)[Ln(dpa)(3)].13H(2)O with Ln = Er(3+), Tm(3+), Yb(3+). AB - The title compounds were synthesized and studied by solution and single-crystal absorption, luminescence, and excitation spectroscopy. The f-f luminescence is induced in the Tm(3+) and Yb(3+) complexes in solution by exciting into the (1)Pi (1)Pi absorptions of the ligand in the UV. A single-configurational coordinate model is proposed to rationalize the nonradiative relaxation step from ligand centered to metal-centered excited states in [Yb(dpa)(3)](3-) (dpa = 2,6 pyridinedicarboxylate). Direct f-f excitation is used in crystals of Na(3)[Tm(dpa)(3)].13H(2)O and Na(3)[Yb(dpa)(3)].13H(2)O to induce f-f luminescence. From low-temperature, high-resolution absorption, luminescence, and excitation spectra, the ligand-field splittings in the relevant states can be determined. It was impossible to induce NIR to VIS upconversion in any of the complexes. This is mainly due to the fact that nonradiative relaxation among the f-f excited states is highly competitive, even in [Yb(dpa)(3)](3-) with an energy gap between (2)F(5/2) and (2)F(7/2) of about 10000 cm(-1). It can be rationalized on the basis of an adapted energy gap law. No luminescence at all could be detected in Na(3)[Er(dpa)(3)].13H(2)O. PMID- 11874338 TI - Synthesis, structure, and photophysical studies of luminescent two- and three dimensional gold-thallium supramolecular arrays. AB - The reactions of tetrahydrofuran solutions of NBu(4)[AuR(2)] (R = C(6)F(5), C(6)Cl(5)) with TlPF(6) and 4,4'-bipyridine lead to the synthesis of the luminescent materials [Tl(bipy)](2)[Au(C(6)F(5))(2)](2) 1 and [Tl(bipy)][Tl(bipy)(0.5)(thf)][Au(C(6)Cl(5))(2)](2) 2 in high yield. The structures of these complexes, as analyzed by X-ray diffraction, consist of planar polymers formed by repetition of Tl-Au-Au-Tl (1) or Tl-Au-Tl'-Au (2) moieties linked through bidentate bridging bipy ligands. In complex 1 these layers are associated via Tl...F contacts between atoms of adjacent planes, whereas in complex 2 each two polymeric layers are linked through additional bridging bipy molecules. Both complexes are strongly luminescent at room temperature and at 77 K in the solid state, losing this characteristic in solution even at high concentrations. The luminescence is attributed to interactions between metal atoms which are strongly affected by their structural dispositions. DFT calculations are in accord with the observed experimental behavior, showing the nature of the orbitals involved in each transition. Detailed analyses reveal a substantial participation of the metals in the transition giving rise to the emission maxima, and also other more energetic bands in which the ligands are involved and which also give rise to these emissions. The obtained theoretical excitation spectra clearly match the experimental results. PMID- 11874339 TI - Anomeric and mesomeric effects in methoxycarbonylsulfenyl chloride, CH(3)OC(O)SCl: an experimental and theoretical study. AB - The molecular structure and conformational properties of methoxycarbonylsulfenyl chloride, CH(3)OC(O)SCl, were determinated in the gas and solid phases by gas electron diffraction, low-temperature X-ray diffraction, and vibrational spectroscopy. Furthermore, quantum chemical calculations were performed. Experimental and theoretical methods result in structures with a planar C-O-C(O) S-Cl skeleton. The electron diffraction intensities are reproduced best with a mixture of 72(8)% syn and 28(8)% anti conformers (S-Cl bond synperiplanar/antiperiplanar with respect to C=O bond) and the O-CH(3) bond synperiplanar with respect to the C=O bond. The syn form is the preferred form and becomes the exclusive form in the crystalline solid at low temperature. This experimental result is reproduced very well by Hartree-Fock approximation and by density functional theory at different levels of theory but not by the MP2/6-311G method, which overestimates the value of DeltaG between the syn and anti conformers. The results are discussed in terms of anomeric effects and a natural bond orbital (NBO) calculation. Photolysis of matrix-isolated CH(3)OC(O)SCl with broad-band UV-visible irradiation produces an interconversion of the conformers, and the concomitant decomposition leads to formation of OCS and CO molecules. PMID- 11874340 TI - Electrochemical, spectroscopic, structural, and magnetic characterization of the reduced and protonated alpha-Dawson anions in [Fe(eta(5) C(5)Me(5))(2)](5)[HS(2)Mo(18)O(62)].3HCONMe(2).2Et(2)O and [NBu(4)](5)[HS(2)Mo(18)O(62)].2H(2)O(1). AB - Reaction of excess Fe(cp)(2) (cp = eta(5)-C(5)Me(5)) dissolved in Et(2)O with [NHex(4)](4)[S(2)Mo(18)O(62)] in acetonitrile, followed by recrystallization of the precipitated solid from N,N'-dimethylformamide (DMF), leads to isolation of the complex [Fe(cp)(2)](5)[HS(2)Mo(18)O(62)].3DMF.2Et(2)O. The solid has been characterized by microanalysis, by voltammetric analysis, by (1)H NMR, diffuse reflectance infrared, EPR, and Mossbauer spectroscopies, and by temperature dependent magnetic susceptibility measurements. The data are consistent with the presence of a paramagnetic [Fe(cp)(2)](+) cation and a diamagnetic two-electron reduced [HS(2)Mo(18)O(62)](5-) anion. The related salt [NBu(4)](5)[HS(2)Mo(18)O(62)].2H(2)O crystallizes in space group C2/c with a = 25.1255(3) A, b = 15.4110(2) A, c = 35.8646(4) A, beta = 105.9381(4), V = 13353.3(3) A(3), and Z = 4. The (2 e(-), 1 H(+))-reduced anion exists as the alpha-Dawson isomer, and its structure may be compared with those of the oxidized and (4 e(-), 3 H(+))-reduced anions as they exist in [NEt(4)](4)[S(2)Mo(18)O(62)].MeCN and [NBu(4)](5)[H(3)S(2)Mo(18)O(62)].4MeCN, respectively. Overall, the anion expands significantly upon the addition of two and then four electrons. However, the Mo...Mo distances along the bonds which connect the two equatorial belts decrease in the order 3.801, 3.780, and 3.736 A, making these distances the shortest for the three inequivalent sets of corner sharing octahedra in each anion. This is consistent with the two or four added electrons localizing essentially in molecular orbitals which are bondiing with respect to interactions between the belts. PMID- 11874341 TI - Molybdenum and tungsten complexes of sulfene (thioformaldehyde S,S-dioxide). AB - Propionitrile complexes fac-[M(CO)(3)(P-P)(NCEt)] (M = Mo (3), W (4); P-P = Ph(2)PCH(2)PPh(2) (a), Ph(2)PC(2)H(4)PPh(2) (b), Ph(2)PC(3)H(6)PPh(2) (c), (S,S) Ph(2)PCHMeCHMePPh(2) (d), Fe(C(5)H(4)PPh(2))(2) (e)) were synthesized from [M(CO)(3)(NCEt)(3)] and the corresponding diphosphine. Reactions of 3 and 4 with sulfur dioxide initially gave complexes fac-[M(CO)(3)(P-P)(eta(2)-SO(2))] (M = Mo (5), W (6)), which slowly isomerized to mer-[M(CO)(3)(P-P)(eta(1)-SO(2))] (M = Mo (7), W (8)). The structures of 7b and 8b were determined by X-ray crystallography. Both compounds are isostructural (monoclinic, space group P2(1)/n (No. 14)) with almost identical unit cell dimensions (7b, a = 14.511(5) A, b = 12.797(2) A, c = 16.476(6) A, beta = 115.92(2); 8b, a = 14.478(8) A, b = 12.794(3) A, c = 16.442(9) A, beta = 116.01(2)) and molecular geometries. Treatment of either fac-[M(CO)(3)(P-P)(eta(2)-SO(2))] or mer-[M(CO)(3)(P P)(eta(1)-SO(2))] with diazomethane yielded the sulfene complexes mer-[M(CO)(3)(P P)(eta(2)-CH(2)SO(2))] (M = Mo (9), W (10)). The structure of 10a was determined crystallographically: monoclinic, space group P2(1)/n (No. 14), a = 11.719(2) A, b = 17.392(4) A, c = 13.441(3) A, beta = 95.58(2). The tungsten atom resides in the center of a distorted pentagonal bipyramid. The sulfene ligand occupies two adjacent equatorial sites with the bond distances W-C, 2.322(13) A, W-S, 2.353(3) A, and S-C, 1.721(12) A. The latter equals the S-C single bond distance in thiirane S,S-dioxide, indicating a high degree of charge density transfer into the LUMO of the sulfene ligand. PMID- 11874342 TI - High-spin metal complexes containing a ferromagnetically coupled tris(semiquinone) ligand. AB - The tris-bidentate ligand 1,3,5-tris(5'-tert-butyl-3',4'-dihydroxyphenyl)benzene ((TBCat)(3)Ph) was synthesized. The reaction of this molecule in basic solution with two paramagnetic acceptors, i.e., a nickel(II)minus signtetraazamacrocyclic ligand complex (Ni(CTH)) (CTH = dl-5,7,7,12,14,14-hexamethyl-1,4,8,11 tetraazacyclotetradecane) and manganese(II)-hydrotris[3-(4'-cumenyl)-5 methylpyrazolyl]borate (Mn(Tp(Cum,Me))), yielded two complexes whose analytical formulas are consistent with those of trinuclear complexes. Spectroscopic and magnetic measurements suggest that these derivatives contain divalent metal ions coordinated to the tris(semiquinone) form of the ligand. Analysis of the magnetic data shows that the pi-connectivity of the ligand enforces ferromagnetic coupling between the three semiquinone units of the molecule, giving rise to complexes with S = 9/2 (M = Ni(II)) and S = 6 (M = Mn(II)) ground states. The coupling within the tris(semiquinone) unit is quite large (J = -26 cm(-1) for the nickel(II) derivative and J = -40 cm(-1) for the manganese(II) one, using the general exchange Hamiltonian H = sigma J(ij)S(i)S(j)), and it is of the same order of magnitude as that observed in an analogous series of bis(semiquinone) complexes that we recently reported. PMID- 11874343 TI - Metal ion-dependent molecular inclusion chemistry: inclusion of aromatic anions by coordinated 1,4,7,10-tetrakis((S)-2-hydroxy-3-phenoxypropyl)-1,4,7,10 tetraazacyclododecane. AB - The ability of the pendant donor macrocyclic ligand 1,4,7,10-tetrakis((S)-2 hydroxy-3-phenoxypropyl)-1,4,7,10-tetraaza- cyclododecane((S)-thphpc12) (or [Cd((S)-thphpc12)](2+)) to act as a metal ion-dependent receptor for aromatic anions has been investigated in solution and in the solid state. [Cd((S) thphpc12)](2+) adopts a stable conical conformation with a large hydrophobic cavity, which has been shown to contain, via complementary multiple hydrogen bonding, p-nitrophenolate, aromatic carboxylates, p-toluenesulfonate, certain aromatic amino acid anions, phenoxyacetate, and acetate. In the case of p nitrophenolate only, one or two anions can be contained within the receptor cavity. The crystal structure of [Cd((S)-thphpc12)(p-nitrophenolate)(2)] shows a coplanar arrangement of the p-nitrophenolates, where each is retained in the cavity by a pair of hydrogen bonds to cis hydroxyl groups. The crystal structure of the p-aminobenzoate inclusion complex indicates retention of the guest via a pair of hydrogen bonds to each oxygen atom of the carboxylate moiety. The crystal structure of the (L)-phenylalaninate inclusion complex indicates that the amino acid is retained by five hydrogen bonds, two involving the nitrogen atom and three to the oxygen atoms of the carboxylate moiety. Binding constants (10(3) 10(5) M(-1)) for the inclusion of some of the aforementioned anions in [Cd((S) thphpc12)](2+) and related receptors were measured by (1)H NMR titration in DMSO d(6) at 298 K. PMID- 11874345 TI - New diamide-diamine ligands and their zirconium and hafnium dichloride and bis(dimethylamide) complexes. AB - The multigram syntheses of the protio ligands (2 NC(5)H(4))CH(2)N(CH(2)CH(2)NHSiMe(2)R)(2) (R = Me, H(2)N(2)NN' 3; R = (t)Bu, H(2)N(2)NN() 4) are described via reactions of the previously reported (2 NC(5)H(4))CH(2)N(CH(2)CH(2)NH(2))(2) (1). A new synthesis of 1 is reported starting from 2-aminomethylpyridine and N-tosylaziridine, proceeding via (2 NC(5)H(4))CH(2)N(CH(2)CH(2)NHTs)(2) (2). Reaction of H(2)N(2)NN' or H(2)N(2)NN* with (n)BuLi gives good yields of the dilithiated derivatives Li(2)N(2)NN' and Li(2)N(2)NN*. Reaction of H(2)N(2)NN' or H(2)N(2)NN* with [MCl(2)(CH(2)SiMe(3))(2)(Et(2)O)(2)] gives the cis-dichloride complexes [MCl(2)(L)] (L = N(2)NN', M = Zr 7 or Hf 8; L = N(2)NN(), M = Zr 9). The corresponding reactions of H(2)N(2)NN' or H(2)N(2)NN* with [Zr(NMe(2))(4)] afford the bis(dimethylamide) derivatives [Zr(NMe(2))(2)(L)] (L = N(2)NN' 10 or N(2)NN* 11). All of these protonolysis reactions proceed smoothly and in good yields. Attempts to prepare the titanium complexes [Ti(X)(2)(N(2)NN')] (X = Cl or NMe(2)) were unsuccessful. The X-ray crystal structures of (2 NC(5)H(4))CH(2)N(CH(2)CH(2)NHTs)(2).EtOH, [ZrCl(2)(N(2)NN')].0.5C(6)H(6), [Zr(NMe(2))(2)(N(2)NN')], and [Zr(NMe(2))(2)(N(2)NN*)] are reported. PMID- 11874344 TI - Kinetic and equilibria studies of the aquation of the trinuclear platinum phase II anticancer agent [(trans-PtCl(NH(3))(2))(2)(mu-trans Pt(NH(3))(2)(NH(2)(CH(2))(6)NH(2))(2))](4+) (BBR3464). AB - The hydrolysis profile of the bifunctional trinuclear phase II clinical agent [(trans-PtCl(NH(3))(2))(2)(mu-trans-Pt(NH(3))(2)(NH(2)(CH(2))(6)NH(2))(2))](4+) (BBR3464, 1) has been examined using [(1)H,(15)N] heteronuclear single quantum coherence (HSQC) 2D NMR spectroscopy. Reported are estimates of the rate and equilibrium constants for the first and second aquation steps, together with the acid dissociation constant (pK(a1) approximately equal to pK(a2) approximately equal to pK(a3)). The equilibrium constants for the aquation determined by NMR at 298 and 310 K (I = 0.1 M, pH 5.3) are similar, pK(1) = pK(2) = 3.35 +/- 0.04 and 3.42 +/- 0.04, respectively. At lower ionic strength (I = 0.015 M, pH 5.3) the values at 288, 293, and 298 K are pK(1) = pK(2) = 3.63 +/- 0.05. This indicates that the equilibrium is not strongly ionic strength or temperature dependent. The aquation and anation rate constants for the two-step aquation model at 298 K in 0.1 M NaClO(4) (pH 5.3) are k(1) = (7.1 +/- 0.2) x 10(-5) s(-1), k(-1) = 0.158 +/ 0.013 M(-1) s(-1), k(2) = (7.1 +/- 1.5) x 10(-5) s(-1), and k(-2) = 0.16 +/- 0.05 M(-1) s(-1). The rate constants in both directions increase 2-fold with an increase in temperature of 5 K, and rate constants increase with a decrease in solution ionic strength. A pK(a) value of 5.62 plus minus 0.04 was determined for the diaqua species [(trans-Pt(NH(3))(2)(OH(2)))(2)(mu-trans Pt(NH(3))(2)(NH(2)(CH(2))(6)-NH(2))(2))](6+) (3). The speciation profile of 1 under physiological conditions is explored and suggests that the dichloro form predominates. The aquation of 1 in 15 mM phosphate was also examined. No slowing of the initial aquation was observed, but reversible reaction between aquated species and phosphate does occur. PMID- 11874346 TI - Synthesis, structure, and preparative transamination of tetrazinc carbamato complexes having the basic zinc carboxylate structure. AB - The series of tetranuclear zinc(II) carbamato complexes (Zn(4)O)(O(2)CR)(6), (1, R = diethylamino; 2, R = piperidino; 3, R = pyrrolidino) was prepared. Complexes 2 and 3 were crystallographically characterized and shown to have the same tetrahedral Zn(4)O(6+) core. Complex 2 crystallizes in the cubic space group I( )43d, a = 24.0131(5) A, V = 13846.6(5) A(3), R(1976 observed reflections) = 0.0194, and GOF = 1.013. Complex 3 crystallizes in the triclinic space group P( )1, a = 10.3178(6) A, b = 10.6962(6) A, c = 19.5130(11) A, alpha = 81.9070(10), beta = 75.4880(10), gamma = 81.6540(10), V = 2050.4(2) A(3), R(6141 observed reflections) = 0.0334, and GOF = 0.979. NMR spectroscopy was used to show that the (Zn(4)O)L(6) structure was maintained in nonpolar solvents. The complexes reacted with free amine in nonpolar solvents, which resulted in facile conversion of one member of the series to another. For example, reacting 1 with a stoichiometric amount of pyrrolidine in tetrahydrofuran followed by workup resulted in the quantitative formation of 3 with liberation of diethylamine. Formally, this is a transamination metathesis reaction between the diethylcarbamate ligand and pyrrolidine. The reaction is complete within 3 min at room temperature, in marked contrast to the extreme conditions required to effect transamination on organic carbamates. The complexes also undergo a facile transcarboxylation reaction with carbon dioxide which results in scrambling of the carboxyl group of the carbamate ligand with free CO(2), also complete in about 3 min. Both transamination and transcarboxylation reactions are consistent with the intermediacy of free CO(2). However, because of the propensity for the complexes to hydrolyze to liberate CO(2), the role of adventitious moisture in facilitating the reaction cannot presently be rejected. PMID- 11874347 TI - Kinetic evidence for intramolecular proton transfer between nickel and coordinated thiolate. AB - The complexes [Ni(YR)(triphos)]BPh(4) (Y = S, R = Ph or Et or Y = Se, R = Ph; triphos = (Ph(2)PCH(2)CH(2))(2)PPh) have been prepared and characterized, and the X-ray crystal structure of [Ni(SPh)(triphos)]BPh(4) has been solved. In MeCN, [Ni(YR)(triphos)](+) are protonated by [lutH](+) (lut = 2,6-dimethylpyridine) to give [Ni(YHR)(triphos)](2+). Studies on the kinetics of these equilibrium reactions reveal an unexpected difference in the reactivities of [Ni(SPh)(triphos)](+) and [Ni(SEt)(triphos)](+). In both cases, the reactions exhibit a first-order dependence on the concentration of complex. When R = Ph, the dependence on the concentrations of [lutH(+)] and lut is given by k(obs) = k(1)(Ph)[lutH(+)] + k(-1)(Ph)[lut], which is typical of an equilibrium reaction where k(1)(Ph) and k(-1)(Ph) correspond to the forward and back reactions, respectively. Analogous behavior is observed for [Ni(SePh)(triphos)](+). However, for [Ni(SEt)(triphos)](+), the kinetics are more complicated, and k(obs) = (k(1)k(2)[lutH(+)] + (k(-2) + k(2)))/(k(1)[lutH(+)] + k(-1)[lut]), which is indicative of a mechanism involving two coupled equilibria in which the initial protonation of the thiolate is followed by a unimolecular equilibrium reaction that is assumed to involve the formation of an eta(2)-EtS-H ligand. The difference in reactivity between the complexes with alkyl and aryl thiolate ligands is a consequence of the (Ni(triphos))(2+) site "leveling" the basicities of these ligands. The pK(a)'s of the PhSH and EtSH constituents coordinated to the (Ni(triphos))(2+) are 16.0 and 14.6, respectively, whereas the difference in pK(a)'s of free PhSH and EtSH differ by ca. 4 units. The pK(a) of [Ni(SeHPh)(triphos)](+) is 14.4. The more strongly sigma-donating EtS ligand makes the (Ni(triphos))(2+) core sufficiently electron-rich that the basicities of the sulfur and nickel in [Ni(SEt)(triphos)](+) are very similar; therefore, the proton serves as a bridge between the two sites. The relevance of these observations to the proposed mechanisms of nickel-based hydrogenases is discussed. PMID- 11874348 TI - Preparation and properties of the aqua ions [W(4)S(4)(H(2)O)(12)](n+) (n = 5, 6) and crystal structure of (Me(2)NH(2))(6)[W(4)S(4)(NCS)(12)].0.5H(2)O. AB - The [3 + 1] reaction of [W(3)S(4)(H(2)O)(9)](4+) with [W(CO)(6)] in 2 M HCl under hydrothermal conditions (130 degrees C) gives the [W(4)S(4)(H(2)O)(12)](6+) cuboidal cluster, reduction potential 35 mV vs NHE (6+/5+ couple). The reduced form is obtained by controlled potential electrolysis. X-ray crystal structure was determined for (Me(2)NH(2))(6)[W(4)S(4)(NCS)(12)].0.5H(2)O. The W-W and W-S bond lengths are 2.840 and 2.379 A, respectively. PMID- 11874349 TI - Carboxylate and alkyl carbonate coordination at the hydrophobic binding site of redox-active dicobalt amine thiophenolate complexes. AB - A series of new dicobalt complexes of the permethylated macrocyclic hexaamine dithiophenolate ligand H(2)L(Me) have been prepared and investigated in the context of ligand binding and oxidation state changes. The octadentate ligand is an effective dinucleating ligand that supports the formation of bioctahedral complexes with a central N(3)Co(mu-SR)(2)(mu-X)CoN(3) core structure, leaving a free bridging position X for the coordination of the substrates. The acetato- and cinnamato-bridged complexes [(L(Me))Co(II)(2)(mu-O(2)CMe)](+) (2) and [(L(Me))Co(II)(2)(mu-O(2)CCH=CHPh)](+) (5) were prepared by reaction of the mu-Cl complex [(L(Me))Co(II)(2)(mu-Cl)](+) (1) with the corresponding sodium carboxylates in methanol. The electrochemical properties of these and of the methyl carbonate complex [(L(Me))Co(II)(2)(mu-O(2)COMe)](+) (8) were also investigated. All complexes undergo two stepwise oxidations at ca. E(1)(1/2) = +0.22 and at E(2)(1/2) = ca. +0.60 V vs SCE, affording the mixed-valent complexes [(L(Me))Co(II)Co(III)(mu-O(2)CR)](2+) (3, 6, 9) and the fully oxidized Co(III)Co(III) forms [(L(Me))Co(III)(2)(mu-O(2)CR)](3+) (4, 7, 10), respectively. Compounds 3, 6, 9 and 4, 7, 10 refer to acetato-, cinnamato-, and methylcarbonato species, respectively. The Co(II)Co(III) compounds were prepared by comproportionation of the respective Co(II)(2) and Co(III)(2) compounds. The Co(III)Co(III) species were prepared by bromine oxidation of the Co(II)Co(II) forms. The crystal structures of complexes 2.BPh(4).MeCN, 3.(I(3))(2), 5.BPh(4).2MeCN, 6.(ClO(4))(2).EtOH, 7.(ClO(4))(3).MeCN.(H(2)O)(3), and 9.(ClO(4))(2).(MeOH)(2).H(2)O were determined by single-crystal X-ray crystallography at 210 K. The oxidations occur without gross structural changes of the parent complexes. The Co(II)Co(III) complexes are composed of high-spin Co(II) (d(7)) and low-spin Co(III) (d(6)) ions. The Co(III)Co(III) complexes are diamagnetic. The oxidation reactions affect the binding mode of the substrates. In the Co(II)(2) and Co(II)Co(III) forms the carboxylates bridge the two Co(2+) ions in a symmetric mu-1,3 fashion with uniform C-O bond distances, whereas asymmetric bridging modes, with one short C=O and one long C-O distance, are adopted in the fully oxidized species. This is consistent with the observed shifts in vibrational frequencies for nu(as)(C-O) and nu(s)(C-O) across the series. PMID- 11874350 TI - First structurally characterized silver(I) derivatives with nonfluorinated beta diketones. AB - Synthetic, spectroscopic, and single-crystal X-ray structural studies of diverse complexes of silver(I) acylpyrazolonate salts AgQ(R') (QH = 1-phenyl-3-methyl-4 R'(C=O)-pyrazol-5-one; Q(1), R = Ph; Q(2), R' = CF(3); Q(3), R' = Me) with neutral ligands L = unidentate PR(3) (R = Ph, o-tolyl, cyclohexyl) and Hmimt (1 methyl-2-mercaptoimidazole) and bidentate dppe (Ph(2)P(CH(2))(2)PPh(2)) and trimen (N,N,N'-trimethylethylenediamine) define the donor capability of the anionic Q(R') ligand in a variety of roles. In the free ligand Q(3)H (which crystallizes in the monoclinic space group C2/c (no. 15), Z = 8, unit cell parameters a = 17.981(6) A, b = 5.0641(4) A, c = 24.271(6) A, and beta = 99.67(2)), the acidic OH group hydrogen-bonds intramolecularly to the adjacent pyrazolone oxygen, i.e., the two oxygen atoms are cis, true of the other Q(R') species structurally characterized here in their anionic complexed forms, in which they chelate the silver in the usual beta-diketonate manner, but not of the free anion, found in the array [Ag(Ph(3)P)(Hmimt)(2)](Q(1)) (triclinic space group P(-)1(no. 2), Z = 2, unit cell parameters a = 11.553(1) A, b = 11.943(1) A, c = 15.479(2) A, alpha = 74.829(2), beta = 76.094(2), and gamma = 78.185(2)), or [Ag(trimen)Q(1)] (monoclinic space group P2(1)/c (no. 14), Z = 4, unit cell parameters a = 7.982(1) A, b = 12.299(2) A, c = 21.507(3) A, and beta = 95.119(3)), which forms an infinite one-dimensional polymer string, Q(1) linking successive silver(I) atoms by coordination by way of the unsubstituted nitrogen and the pyrazolonate oxygen. In all [Ag(R(3)P)(2)(chelate-Q(1))] (R = Ph, Cy) complexes, P(2)Ag(O,O') arrays are found (R = Ph, monoclinic space group C2/c (no. 15), Z = 8, unit cell parameters a = 16.193(8) A, b = 13.859(7) A, c = 39.306(7) A, and beta = 100.02(3); R = Cy, triclinic space group P(-)1(no. 2), Z = 2, unit cell parameters a = 10.4655(9) A, b = 12.079(1) A, c = 22.804(2) A, alpha = 104.872(2), beta = 95.180(2), and gamma = 104.144(2)), also true of [Ag(Ph(3)P)(2)(O,O'-Q(2))] (triclinic space group P(-)1(no. 2), Z = 2, unit cell parameters a = 10.672(2) A, b = 10.710(2) A, c = 18.713(3) A, alpha = 87.573(2), beta = 80.972(2), and gamma = 81.734(2)), whereas [Ag(o-tol(3)P)Q(1)] (monoclinic space group P2(1)/c (no. 14), Z = 2 dimers, unit cell parameters a = 11.8221(6) A, b = 13.2601(6) A, c = 20.5141(10) A, and beta = 91.758(1)) exists as a dinuclear species containing two AgO(2)NP units where the acylpyrazolonate is coordinated in a bridging O,O'-Q-Nfashion. Silver atoms are four-coordinate in all except the Hmimt complex. PMID- 11874351 TI - Photophysical effect of the coordination of water by ruthenium(II) bipyridyl complexes containing hemilabile phosphine-ether ligands. AB - A substantial concentration-dependent red shift of the absorption and emission spectra (77 K) of [Ru(bpy)(2)(POMe-P,O)](2+) (1) (POMe = (2 methoxyphenyl)diphenylphosphine) is reported. NMR experiments show this shift to be due to equilibration of 1 with an aquo complex (1b) (K(eff) = (6 +/- 3) x 10( 3)) that forms upon displacement of the coordinated ether in the hemilabile POMe ligand. The excited-state lifetimes of 1 and 1b at 77 K in solid 2:1 ethanol/acetone solution are tau = 2.13 +/- 0.02 and 1.95 +/- 0.02 mus, respectively. The preparation and X-ray crystal structure of a related complex, [Ru(bpy)(2)(PO(i)Pr-P)(OH(2))](PF(6))(2) (2b) (PO(i)Pr-P = (2-(2 propoxy)phenyl)diphenylphosphine), is also reported. In solution, this species exists as an equilibrium mixture of complexes that cannot be readily separated. This species also has concentration-dependent absorption spectra in 2:1 ethanol/acetone solution, with a significant red shift (20 nm) at lower concentrations. PMID- 11874352 TI - Synthesis, properties, and crystal structure of a novel anthracene-bridged molybdenum-zinc porphyrin dimer. AB - The synthesis and properties of novel anthracene-bridged porphyrin dimers having an oxomolybdenum(V) porphyrin unit, H(2)(DPA)[Mo(V)O(OMe)] (1) and (DPA)[Mo(V)O(OMe)][Zn(II)(MeOH)] (2), and the relevant monomer porphyrin complexes Mo(V)O(MPP)OMe (3) and Zn(II)(MPP) (4) are presented. An oxomolybdenum(V) unit was introduced into one of the two porphyrins in DPA to give 1, which has a free-base porphyrin unit. By introducing a zinc(II) ion to the free-base part, a mixed-metal complex of 2 was prepared and isolated. The structure of 2 was analyzed by X-ray crystallography (2.(7)/(6)CH(2)Cl(2), triclinic, P(-)1 (no. 2), a = 15.2854(12) A, b = 19.9640(15) A, c = 13.6915(12) A, alpha = 90.968(3), beta = 113.108(4), gamma = 96.501(4), Z = 2, R1 = 9.9, wR2 = 19.2). The structure of 2 demonstrated that a methanol is stably coordinated to the Zn(II) ion with the aid of a hydrogen bond to the methoxo ligand on the Mo(V) ion in the binding pocket of DPA. The electrochemical measurements of 2 suggested that the methanol was kept in the pocket of DPA in solution even at the reduced state of the molybdenum ion. PMID- 11874353 TI - Hydrothermal syntheses, structures, and properties of the new uranyl selenites Ag(2)(UO(2))(SeO(3))(2), M[(UO(2))(HSeO(3))(SeO(3))] (M = K, Rb, Cs, Tl), and Pb(UO(2))(SeO(3))(2). AB - The transition metal, alkali metal, and main group uranyl selenites, Ag(2)(UO(2))(SeO(3))(2) (1), K[(UO(2))(HSeO(3))(SeO(3))] (2), Rb[(UO(2))(HSeO(3))(SeO(3))] (3), Cs[(UO(2))(HSeO(3))(SeO(3))] (4), Tl[(UO(2))(HSeO(3))(SeO(3))] (5), and Pb(UO(2))(SeO(3))(2) (6), have been prepared from the hydrothermal reactions of AgNO(3), KCl, RbCl, CsCl, TlCl, or Pb(NO(3))(2) with UO(3) and SeO(2) at 180 degrees C for 3 d. The structures of 1 5 contain similar [(UO(2))(SeO(3))(2)](2-) sheets constructed from pentagonal bipyramidal UO(7) units that are joined by bridging SeO(3)(2-) anions. In 1, the selenite oxo ligands that are not utilized within the layers coordinate the Ag(+) cations to create a three-dimensional network structure. In 2-5, half of the selenite ligands are monoprotonated to yield a layer composition of [(UO(2))(HSeO(3))(SeO(3))](1-), and coordination of the K(+), Rb(+), Cs(+), and Tl(+) cations occurs through long ionic contacts. The structure of 6 contains a uranyl selenite layered substructure that differs substantially from those in 1-5 because the selenite anions adopt both bridging and chelating binding modes to the uranyl centers. Furthermore, the Pb(2+) cations form strong covalent bonds with these anions creating a three-dimensional framework. These cations occur as distorted square pyramidal PbO(5) units with stereochemically active lone pairs of electrons. These polyhedra align along the c-axis to create a polar structure. Second-harmonic generation (SHG) measurements revealed a response of 5x alpha quartz for 6. The diffuse reflectance spectrum of 6 shows optical transitions at 330 and 440 nm. The trailing off of the 440 nm transition to longer wavelengths is responsible for the orange coloration of 6. PMID- 11874354 TI - Novel organotellurium(IV) diazides and triazides. AB - New series dialkyltellurium(IV) diazides R(2)Te(N(3))(2) (R = CH(3) (6), C(2)H(5) (7), n-C(3)H(7) (8), i-C(3)H(7) (9), c-C(6)H(11) (10)) and alkyl/aryltellurium(IV) triazides R'Te(N(3))(3) (R' = CH(3) (11), C(2)H(5) (12), n-C(3)H(7) (13), i-C(3)H(7) (14), C(6)H(5) (15), 2,4,6-(CH(3))(3)C(6)H(2) (16)) were synthesized by the straightforward substitution of fluorine atoms in the corresponding tellurium difluorides, or trifluorides respectively, with trimethylsilyl azide. In addition to standard characterization methods, the first crystal structures of covalent organotellurium(IV) triazides 12, 13, 14, and 16 have been determined. Ethyltellurium(IV) triazide, C(2)H(5)Te(N(3))(3) (12), crystallizes in the monoclinic space group P2(1)/n, a = 8.4530(2) A, b = 7.9094(2) A, c = 12.6288(3) A, beta = 91.876(1). n-Propyltellurium(IV) triazide, n-C(3)H(7)Te(N(3))(3) (13), crystallizes in the monoclinic space group P2(1)/n as well, a = 8.7999(2) A, b = 7.9674(2) A, c = 13.2334(3) A, beta = 94.626(1). Isopropyltellurium(IV) triazide, i-C(3)H(7)Te(N(3))(3) (14), crystallizes in the monoclinic space group C2/c, a = 20.058(2) A, b = 6.9620(3) A, c = 15.030(1) A, beta = 114.260(9). Mesityltellurium(IV) triazide, 2,4,6 (CH(3))(3)C(6)H(2)Te(N(3))(3) (16), crystallizes monoclinic as well; the space group is P2(1)/c, a = 7.5503(6) A, b = 23.581(1) A, c = 7.5094(6) A, beta = 91.295(9). The structures and vibrational frequencies of the methyl derivatives dimethyltellurium(IV) diazide (6) and methyltellurium(IV) triazide (11) have been calculated by density functional theory methods and were compared with the experimental metric parameters and vibrational data. PMID- 11874355 TI - Metal scrambling in the trinuclear (Pt(2)Se(2)M)(M = Pt, Pd, Au) system using an electrospray mass spectrometry (ESMS) directed synthetic methodology; isolation and crystallographic characterization of (Pt(2)(mu(3) Se)(2)(PPh(3))(4)[Pt(cod)])(PF(6))(2) and (Pt(mu(3) Se)(2)(PPh(3))(2)[Pt(cod)](2))(PF(6))(2) (cod = cyclo-octa-1,5-diene). AB - Pt(2)(mu-Se)(2)(PPh(3))(4) reacts with PtCl(2)(cod) to give (Pt(2)(mu(3) Se)(2)(PPh(3))(4)[Pt(cod)])(2+) and an unexpected cod-rich product that arises from metal scrambling, viz. (Pt(mu(3)-Se)(2)(PPh(3))(2)[Pt(cod)](2))(2+). The formation of these species was detected and followed by electrospray mass spectrometry (ESMS) and subsequently verified by batch synthesis and crystallographic characterization. Other metal-scrambled aggregate products were successfully detected. PMID- 11874356 TI - Tuning of optical band gaps: syntheses, structures, magnetic properties, and optical properties of CsLnZnSe(3) (Ln = Sm, Tb, Dy, Ho, Er, Tm, Yb, and Y). AB - Eight new quaternary selenides CsSmZnSe(3), CsTbZnSe(3), CsDyZnSe(3), CsHoZnSe(3,) CsErZnSe(3), CsTmZnSe(3), CsYbZnSe(3), and CsYZnSe(3) have been synthesized with the use of high-temperature solid-state experimental methods. These compounds are isostructural with KZrCuS(3), crystallizing with four formula units in the orthorhombic space group Cmcm. The structure of these CsLnZnSe(3) compounds is composed of [LnZnSe(3)(-)] layers separated by Cs atoms. The Ln atom is octahedrally coordinated by six Se atoms, the Zn atom is tetrahedrally coordinated by four Se atoms, and the Cs atom is coordinated by a bicapped trigonal prism of eight Se atoms. Because there are no Se-Se bonds in the structure, the oxidation state of Cs is 1+, that of Ln is 3+, and that of Zn is 2+. CsYbZnSe(3) exhibits an antiferromagnetic transition at 11 K, whereas CsSmZnSe(3) does not follow a Curie-Weiss law. The remaining rare-earth compounds are paramagnetic, and the calculated effective magnetic moments of the rare-earth ions agree well with their theoretical values. Optical absorption data on face indexed single crystals of CsSmZnSe(3), CsErZnSe(3), CsYbZnSe(3), and CsYZnSe(3) demonstrate that the optical band gap changes by more than 0.75 eV with the composition and by as much as 0.20 eV with the crystal orientation. The optical band gaps range from 2.63 eV (CsSmZnSe(3), CsErZnSe(3)) to 1.93 eV (CsYbZnSe(3)) for the (010) crystal face and 2.56 eV (CsErZnSe(3)) to 1.88 eV (CsYbZnSe(3)) for the (001) crystal face. The difference in the optical band gap of the (010) face vs the (001) face varies from +0.05 eV (CsYbZnSe(3)) to +0.20 eV (CsSmZnSe(3)). PMID- 11874357 TI - Coordination chemistry of silver(I) with the nitrogen-bridged ligands (C(6)H(5))(2)PN(H)P(C(6)H(5))(2) and (C(6)H(5))(2)PN(CH(3))P(C(6)H(5))(2): the effect of alkylating the nitrogen bridge on ligand bridging versus chelating behavior. AB - The coordination chemistry of silver(I) with the nitrogen-bridged ligands (C(6)H(5))(2)PN(R)P(C(6)H(5))(2) [R = H (dppa); R = CH(3) (dppma)] has been investigated by (31)P NMR and electrospray mass spectrometry (ESMS). Species observed by (31)P NMR include Ag(2)(mu-dppa)(2+), Ag(2)(mu-dppa)(2)(2+), Ag(2)(mu dppa)(3)(2+), Ag(2)(mu-dppma)(2+), Ag(2)(mu-dppma)(2)(2+), and Ag(eta(2) dppma)(2)(+). Species observed by ESMS at low cone voltages were Ag(2)(dppa)(2)(2+), Ag(2)(dppa)(3)(2+), Ag(2)(dppma)(2)(2+), and Ag(dppma)(2)(+). (C(6)H(5))(2)PN(CH(3))P(C(6)H(5))(2) showed a strong tendency to chelate, while (C(6)H(5))(2)PN(H)P(C(6)H(5))(2) preferred to bridge. Differences in the bridging versus chelating behavior of the ligands are assigned to the Thorpe-Ingold effect, where the methyl group on nitrogen sterically interacts with the phenyl groups on phosphorus. The crystal structure of the three-coordinate dinuclear silver(I) complex (Ag(2)[(C(6)H(5))(2)PN(H)P(C(6)H(5))(2)](3))(BF(4))(2) has been determined. Bond distances include Ag-Ag = 2.812(1) A, Ag(1)-P(av) = 2.492(3) A, and Ag(2)-P(av) = 2.509(3) A. The compound crystallizes in the monoclinic space group Cc at 294 K, with a = 18.102(4)(o), Z = 4, V = 7261(3) A(3), R = 0.0503, and R(W) = 0.0670. PMID- 11874358 TI - Photochemical or thermal chelate exchange in the ruthenium coordination sphere of complexes of the Ru(phen)(2)L family (L = diimine or dinitrile ligands). AB - Complexes of the type Ru(phen)(2)L(2+), where L is a substituted bipyridine family member, have been prepared, and their photochemical substitution reactions have been investigated. In the presence of a bis-benzonitrile derivative, acting as a bidentate chelate, photoexpulsion of L is performed under the action of visible light, with quantitative formation of new complexes of the type Ru(phen)(2)L'(2+) (L' = bis-nitrile ligand). Several complexes have been characterized by X-ray crystallography. In particular, the bis-benzonitrile complexes could be crystallized, and the structure of these compounds, containing a 13-, 14-, or 15-membered metal incorporating ring, was obtained. By heating Ru(phen)(2)L'(2+) with a bipy derivative in refluxing ethylene glycol, quantitative formation of the starting complex [Ru(phen)(2)L(2+)] was carried out. The present series of compounds presents properties that could be profitably used in the design and construction of multicomponent systems acting as photochemically driven molecular machines. PMID- 11874359 TI - Structural investigations of palladium(II) and platinum(II) complexes of salicylhydroxamic acid. AB - Complexes of salicylhydroxamic acid (shaH) with palladium(II) and platinum(II) were investigated. The synthesis of [Pt(sha)(2)] was attempted via a number of methods, and ultimately (1)H NMR investigations revealed that salicylhydroxamate would not coordinate to chloro complexes of platinum(II). However, [Pt(sha H)(PPh(3))(2)] was successfully synthesized and the crystal structure determined (orthorhombic, space group Pca2(1) a = 17.9325(19) A, b = 11.3102(12) A, c = 18.2829(19) A, Z = 4, R = 0.0224). The sha binds via an [O,O] binding mode, in its hydroximate form. In contrast the palladium complex [Pd(sha)(2)] was readily synthesized and crystallized as [Pd(sha)(2)](DMF)(4) in the triclinic space group P(-)1,a = 7.066(1) A, b = 9.842(2) A, c = 12.385(2) A, alpha = 99.213(3)(o), beta = 90.669(3), gamma = 109.767(3)(o), Z = 1, R = 0.037. The unexpected [N,O'] binding mode of the salicylhydroxamate ligand in [Pd(sha)(2)] prompted investigation of the stability of a number of binding modes of salicylhydroxamic acid in [M(sha)(2)] (M = Pd, Pt) by density functional theory, using the B3LYP hybrid functional at the 6-311G* level of theory. Geometry optimizations were carried out for various binding modes of the ligands and their relative energies established. It was found that the [N,O'] mode gave the more stable complex, in accord with experimental observations. Stabilization of hydroxamate binding to platinum is evidently afforded by soft ligands lying trans to them. PMID- 11874360 TI - Electronic energy self-exchange with macrocyclic chromium(III) complexes. AB - The luminescence lifetimes of N-deuterated Cr(III) complexes of macrocyclic tetraamine ligands, trans-CrN(4)X(2)(n)()(+), are substantially longer than those of their undeuterated counterparts in room temperature solution. Thus, excited state emission quenching of the longer lived species by the shorter lived species may be studied by analyzing the decay profile following pulsed excitation. Flash photolysis experiments were carried out for three deuterated/undeuterated pairs of trans-CrN(4)X(2)(n)()(+) complexes (where X = CN-, NH(3), and F-). For the trans-Cr(cyclam)(CN)(2)(+) system in H(2)O, it was determined that energy transfer was occurring between the deuterated and undeuterated species. Although the rate constant of energy transfer was too fast to measure explicitly, it could be bracketed as k(et) >>7 x 10(6) M(-1) s(-1). For this reaction it was possible to measure an equilibrium constant which was very near 1.0. For trans Cr(cyclam)(NH(3))(2)(3+) in DMSO, it was also established that energy transfer was occurring and rate constants of 2.4 x 10(6) M(-1) s(-1) (mu = 0.1) and 9.7 x 10(6) M(-1) s(-1) (mu = 1.0) were determined by a Stern-Volmer analysis. For trans-Cr(tet a)F(2+) in H(2)O, no energy transfer was observed, which implies that the rate constant is <<3 x 10(5) M(-1) s(-1). Because these energy-transfer reactions represent self-exchange energy transfer and are thus thermoneutral, we are able to analyze the results using Marcus theory and draw some conclusions about the relative importance of nuclear reorganization and electronic factors in the overall rate. PMID- 11874361 TI - Microwave spectra, geometries, and hyperfine constants of OCAgX (X = F, Cl, Br). AB - A pulsed jet cavity Fourier transform microwave spectrometer has been used to measure the rotational spectra of OCAgX (X = F, Cl, Br) in the frequency range 5 22 GHz. Metal atoms were generated via laser ablation and were allowed to react with CO and a halide precursor, prior to stabilization of the products within a supersonic jet of argon. These are the first experimental observations of OCAgF and OCAgBr, and the first high resolution spectroscopic study of OCAgCl. All three molecules are linear. Accurately determined rotational constants have been used to evaluate the various internuclear distances, which are found to be consistent with trends established for OCAuX and OCCuX species. The C-O distances are short, and the M-C distances are significantly longer than those in other molecules containing a metal-carbonyl bond. Precise values of centrifugal distortion constants and halogen nuclear quadrupole coupling constants have also been determined. The coupling constants are compared with the results of previous studies of OCCuX and OCAuX and are used to infer trends in the electron distributions of the molecules. Ab initio calculations have been performed and employed to predict the geometries, vibrational frequencies, and Mulliken valence orbital populations of the various species. PMID- 11874362 TI - Syntheses and structures of P-anilino-P-chalcogeno- and P-anilino-P iminodiazasilaphosphetidines and their group 12 and 13 metal compounds. AB - The P-anilino-P-chalcogeno(imino)diazasilaphosphetidines [Me(2)Si(mu N(t)Bu)(2)P=E(NHPh)] (E = O (3), S (4), Se (5), N-p-tolyl (6)) were synthesized by oxidizing the P-anilinodiazasilaphosphetidine [Me(2)Si(N(t)Bu)(2)P(NHPh)] (2) with cumene hydroperoxide, sulfur, selenium, and p-tolyl azide, respectively. The lithium salt of 4 reacted with thallium monochloride to produce ([Me(2)Si(mu N(t)Bu)(2)P=S(NPh)-kappaN-kappaS]Tl)(7), which features a two-coordinate thallium atom. Treatment of 4-6 with AlMe(3) gave the monoligand dimethylaluminum complexes ([Me(2)Si(mu-N(t)Bu)(2)P=E(NPh)-kappaN-kappaE]AlMe(2)) (E = S (8), Se (9), N-p-tolyl (10)), respectively. In these complexes the aluminum atom is tetrahedrally coordinated by one chelating ligand and two methyl groups, as a single-crystal X-ray analysis of 8 showed. A 2 equiv amount of 4-6 reacted with diethylzinc to produce the homoleptic diligand complexes ([Me(2)Si(mu N(t)Bu)(2)P=E(NPh)-kappaN-kappaE](2)Zn)(E = S (11), Se (12), N-p-tolyl (13)). A crystal-structure analysis of 11 revealed a linear tetraspirocycle with a tetrahedrally coordinated, central zinc atom. PMID- 11874363 TI - Charge-transfer studies of iron cyano compounds bound to nanocrystalline TiO(2) surfaces. AB - Nanocrystalline (anatase) titanium dioxide films have been sensitized to visible light with K(4)[Fe(CN)(6)] and Na(2)[Fe(LL)(CN)(4)], where LL = bpy (2,2' bipyridine), dmb (4,4'-dimethyl-2,2'-bipyridine), or dpb (4,4'-diphenyl-2,2' bipyridine). Coordination of Fe(CN)(6)(4-) to the TiO(2) surface results in the appearance of a broad absorption band (fwhm approximately 8200 cm(-1)) centered at 23800 +/- 400 cm(-1) assigned to an Fe(II)-->TiO(2) metal-to-particle charge transfer (MPCT) band. The absorption spectra of Fe(LL)(CN)(4)(2-) compounds anchored to TiO(2) are well modeled by a sum of metal-to-ligand charge-transfer (MLCT) bands and a MPCT band. Pulsed light excitation (417 or 532 nm, approximately 8 ns fwhm, approximately 2-15 mJ/pulse) results in the immediate appearance of absorption difference spectra assigned to an interfacial charge separated state [TiO(2)(e(-)), Fe(III)], k(inj) > 10(8) s(-1). Charge recombination is well described by a second-order equal concentration kinetic model and requires milliseconds for completion. A model is proposed wherein sensitization of Fe(LL)(CN)(4)(2-)/TiO(2) occurs by MPCT and MLCT pathways, the quantum yield for the latter being dependent on environment. The solvatochromism of the materials allows the reorganization energies associated with charge transfer to be quantified. The photocurrent efficiencies of the sensitized materials are also reported. PMID- 11874364 TI - Trinuclear pyrazine-bridged ruthenium complexes: syntheses, electrochemistry, NIR Vis spectra, and their interpretation in terms of a 5-orbital-3-parameter model. AB - A study of absorption spectra in the near-infrared (NIR) and visible (vis) regions of trinuclear Ru complexes containing pyrazine (pyz) as bridging ligand, trans-[(Ru(NH(3))(5)pyz)(2)Ru(NH(3))(4)](m+)(m = 6-9), is reported. The spectra were recorded on aqueous solutions containing the described species formed in situ by stoichiometric additions of a standard solution of Ce(SO(4))(2). They were interpreted in terms of a simple 5-orbital-3-parameter model which includes the effects of d-pi interaction and electronic correlation. The model is shown to account for the observed NIR-vis spectra of the complex ions. The 6+ parent species was synthesized by an improved literature method and fully characterized. The novel 8+ complex was also prepared and characterized. The 9+ ion was established to be slowly reduced by water, with dioxygen formation. Electrochemical (CV and DPV) studies were performed on the trinuclear 6+ complex, as well as on its constituent fragments [Ru(NH(3))(5)(pyz)](2+) and trans [Ru(NH(3))(4)(pyz)(2)](2+). PMID- 11874365 TI - Oxorhenium(V) dithiolates catalyze the oxidation by tert-butyl hydroperoxide of sulfoxides and sulfides, including 4,6-dimethyldibenzothiophene. AB - tert-Butyl hydroperoxide (TBHP) efficiently converts a wide variety of sulfides to sulfoxides and sulfones. The method offers the advantage that one product or the other can be obtained in high purity by a modest variation of conditions. The reactions occur smoothly at 25minus sign50 C in chloroform and, to the extent studied, in toluene and methylene chloride. A catalyst is required; the most extensively studied was MeReO(mtp)PPh(3), 1, where mtpH(2) is 2 (mercaptomethyl)thiophenol. Other chelating dithiolate ligands can be used with comparable results. These oxidations were tested for dialkyl, alkylminus signaryl, and diaryl sulfides; thiophenes; and thianthrene. Even the "hard" sulfide, 4,6-dimethyldibenzothiophene (DMDBT) was quantitatively oxidized to the dioxide with TBHP:DMDBT 3.0-3.5 and 0.05-3.8 mol % 1. The mechanism was explored in kinetics studies carried out only for methyl tolyl sulfide. The product buildup curve was complex, with an induction period followed by a rapid growth phase. The kinetic data could be modeled adequately but not perfectly by allowing five rate constants to refine. Their values are consistent with the chemical sense of the mechanism. PMID- 11874366 TI - Synthesis, characterization, electrochemistry, electronic structure, and isomerization of mononuclear oxo-molybdenum(V) complexes: the serine gate hypothesis in the function of DMSO reductases. AB - Crystal structures of DMSO reductases isolated from two different sources and the crystal structure of related trimethylamine-N-oxide reductase indicate that the angle between the terminal oxo atom on the molybdenum and the serinato oxygen varies significantly. To understand the significance of this angular variation, we have synthesized two isomeric compounds of the heteroscorpionato ligand (L1OH) (cis- and trans-(L1O)Mo(V)OCl(2)), where the phenolic oxygen mimics the serinato oxygen donor. Density functional and semiempirical calculations indicate that the trans isomer is more stable than the cis. The lower stability of the cis isomer can be attributed to two factors. First, a strong antibonding interaction between the phenolic oxygen with molybdenum d(xy) orbital raises the energy of this orbital. Second, the strong trans influence of the terminal oxo group in the trans isomer places the phenol ring, and hence the bulky tertiary butyl group, in a less sterically hindered position. In solution, the cis isomer spontaneously converts to the thermodynamically favorable trans isomer. This geometric transformation follows a first-order process, with an enthalpy of activation of 20 kcal/mol and an entropy of activation of -9 cal/mol K. Computational analysis at the semiempirical level supports a twist mechanism as the most favorable pathway for the geometric transformation. The twist mechanism is further supported by detailed mass spectral data collected in the presence of excess tetraalkylammonium salts. Both the cis and trans isomers exhibit well-defined one electron couples due to the reduction of molybdenum(V) to molybdenum(IV), with the cis isomer being more difficult to reduce. Both isomers also exhibit oxidative couples because of the oxidation of molybdenum(V) to molybdenum(VI), with the cis isomer being easier to oxidize. This electrochemical behavior is consistent with a higher-energy redox orbital in the cis isomer, which has been observed computationally. Collectively, this investigation demonstrates that by changing the O(t)-Mo-O(p) angle, the reduction potential can be modulated. This geometrically controlled modulation may play a gating role in the electron transfer process during the regeneration steps in the catalytic cycle. PMID- 11874367 TI - Nanoscale characterization of redox and acid properties of keggin-type heteropolyacids by scanning tunneling microscopy and tunneling spectroscopy: effect of heteroatom substitution. AB - Nanoscale characterization of acid and redox properties of Keggin-type heteropolyacids (HPAs) with different heteroatoms, H(n)MW(12)O(40) (M = P, Si, B, Co), was carried out by scanning tunneling microscopy (STM) and tunneling spectroscopy (TS) in this study. HPA samples were deposited on highly oriented pyrolytic graphite surfaces to obtain images and tunneling spectra by STM before and after pyridine adsorption. All HPA samples formed well-ordered 2-dimensional arrays on graphite before and after pyridine exposure. NDR (negative differential resistance) peaks were observed in the tunneling spectra. Those measured for fresh HPA samples appeared at less negative voltages with increasing reduction potential of the HPAs and with increases in the electronegativity of the heteroatom, but with decreases in the overall negative charge of the heteropolyanions. These results support the conclusion that more reducible HPA samples show NDR behavior at less negative applied voltages in their tunneling spectra. Introduction of pyridine into the HPA arrays increased the lattice constants of the 2-dimensional HPA arrays by ca. 6 A. Exposure to pyridine also shifted NDR peak voltages of H(n)MW(12)O(40) (M = P, Si, B, Co) samples to less negative values in the TS measurements. The NDR shifts of HPAs obtained before and after pyridine adsorption were correlated with the acid strengths of the HPAs, suggesting that tunneling spectra measured by STM could serve to probe acid properties of HPAs. These results show how one can relate the bulk acid and redox properties of HPAs to surface properties of nanostructured HPA monolayers determined by STM. PMID- 11874368 TI - Density functional theory study of anionic and neutral per-substituted 12-vertex boron cage systems, B(12)X(12)(n-) (n = 2, 1, 0). AB - The 12(12) closomers form a rapidly expanding class of compounds where a 12 vertex cage is surrounded by 12 identical substituents. Density functional theory (B3LYP/6-31G(d)) is used to study a number of these closomers in different states of oxidation (dianion, radical anion, and neutral cages). The cage stability increases as the group electronegativity of the substituent increases. Also, the 12(12) closomer becomes easier to oxidize as the Hammett sigma(p) parameter becomes more negative (electron-donating). As the closomer is oxidized, the size of the cage increases and the B-B distances become more asymmetric. The Raman active breathing mode in the 404-434 cm(-1) range moves to lower frequency as the cage is oxidized, which is caused by removing one or two electrons from a cage bonding molecular orbital. PMID- 11874369 TI - Ligand substitution kinetics of the iron(III) hydroxo dimer with simple inorganic ligands. AB - The kinetics and mechanisms of ligand substitution reactions of the iron(III) hydroxo dimer, Fe(2)(mu-OH)(2)(H(2)O)(8)(4+), with various inorganic ligands were studied by the stopped-flow method at 10.0 or 25.0 C in 1.0 M NaClO(4). The transient formation of the following di- and tetranuclear complexes was confirmed: Fe(2)(OH)SO(4)(3+), Fe(2)(OH)H(2)PO(2)(4+), Fe(2)(OH)HPO(3)(3+), Fe(2)(OH)SeO(3)(3+), and Fe(4)(AsO(4))(OH)(2)(7+). The catalytic effect of arsenic(III) on the hydrolytic reaction of iron(III) was also attributed to the formation of a dinuclear complex at very low concentration levels. Fast formation and subsequent dissociation of the multinuclear species into the corresponding mononuclear complexes (FeL) proceed via parallel reaction paths which, in general, show composite pH dependencies. The appropriate rate laws were established. The reactions of the different ligands occur at very similar rates, though the uninegatively charged singly deprotonated form reacts about 1 order of magnitude faster than the neutral form of the same ligand. The results can conveniently be interpreted in terms of a dissociative interchange mechanism which postulates the formation of an intermediate complex in which the ligand is coordinated to only one Fe(III) center of the hydroxo dimer. In a subsequent fast step, the ligand forms a bridge between the two metal ions by replacing one of the OH groups. The dissociation of the dinuclear complex into FeL most likely proceeds via the same intermediate. PMID- 11874370 TI - Coordination of the neptunyl ion with carbonate ions and water: a theoretical study. AB - The results of a study on the ground-state of monocarbonate, bicarbonate, and tricarbonate complexes of neptunyl using multiconfigurational second-order perturbation theory (CASSCF/CASPT2) are presented. The equilibrium geometries of the complexes corresponding to neptunium in the formal oxidation state (V) have been fully optimized at the CASPT2 level of theory in the presence of an aqueous environment modeled by a reaction field Hamiltonian with a spherical cavity. Some water molecules have been explicitly included in the calculation. This study is consistent with the hypothesis that the monocarbonate complex has a pentacoordinated structure with three water molecules in the first coordination shell and that the bicarbonate complex has a hexacoordinated structure, with two water molecules in the first coordination shell. The typical bond distances are in good agreement with experimental results. The tricarbonate complex was studied with explicit counterions, which resulted in somewhat longer Np-carbonate bond distances than experiment indicates. PMID- 11874371 TI - Effects of first-row substituents on silicon-phosphorus triple bonds. PMID- 11874372 TI - Pentanuclear octacyanotungstate(V)-based molecule with a high spin ground state S= (13/2). PMID- 11874373 TI - Structural model for the Cu(B) site of dopamine beta-hydroxylase: crystal structure of a copper(II) complex showing N(3)OS coordination with an axial sulfur ligation. PMID- 11874374 TI - First cubane-like polysulfidomolybdate: synthesis and crystal structure of Cs(6)[Mo(4)S(23.6)]. PMID- 11874375 TI - Two entities in which a dissociative hypertropia may present after surgery. PMID- 11874376 TI - Further debate regarding surgery for accommodative esotropia. PMID- 11874377 TI - The interface between ophthalmology and optometric vision therapy. AB - Considerable disparity lies between ophthalmologic impressions of optometric vision therapy, and the reality of optometric vision therapy as practiced in the United States. The viewpoint shared by ophthalmology in particular, and the medical field in general, is one that is filtered through organizational policy statements and the isolated experiences of influential individual practitioners. This has resulted in a skewed portrayal of optometric vision therapy. The purpose of this paper is to present a balanced perspective on this subject, and one that should be of assistance in creating an interface between ophthalmology and optometry that better serves the public. PMID- 11874378 TI - Optometric vision therapy and training for learning disabilities and dyslexia: DVD surgery; curing complications of strabismus surgery. PMID- 11874379 TI - A surgical alternative for dissociated vertical deviation based on new pathologic concepts: weakening all four oblique eye muscles. Outcome and results in 9 cases. AB - PURPOSE: To test new and recent theories on the etiology of Dissociated Vertical Deviation (DVD) with an appropriate new alternative surgical technique. METHOD: Nine patients were prospectively selected. The surgical technique used was simultaneous and symmetrical weakening of all 4 oblique eye muscles in order to reduce bilateral cyclotorsion. For analysis, "Statistical Significance" level used was p<0.05. RESULTS: Mean preoperative hypertropia was 17.9 prism diopters (pd) for the right eyes (RE) and 17.7 pd for the left eyes (LE). Mean post surgical deviation achieved was 6.44 pd for the REs and 5.78 pd for the LEs. Statistical analysis (Wilcoxon's Test) showed a p<0.02 for both eyes. In all cases, a symmetrical correction was also obtained. CONCLUSIONS: The hypothesis that the manifest hypertropia seen in patients with DVD is secondary to cyclotorsion, mediated primarily by the oblique muscles, was validated by improving (reducing) the DVD by performing bilateral and symmetrical weakening of all four oblique extraocular muscles. PMID- 11874380 TI - Diagnosis and management of the surgical complication of postoperative "slipped" medial rectus muscle: a new "tendon step test" and outcome/results in 11 cases. AB - OBJECTIVES: To report our experience in eleven patient with a "slipped" medical rectus after prior surgery. We describe the "tendon step test" as the basis for early intraoperative suspicion of a "slipped" muscle. The diagnostic translucent empty capsule is usually identified following careful dissection of the fibrous tissue surrounding the capsule. METHODS: The patients medical records were reviewed and analyzed retrospectively. Preoperatively, adduction had been normal in one case, mild to moderately limited in eight and severely limited in two. Proptosis and widening of palpebral fissure were inconstant features. We had performed the tendon step tests intraoperatively and proceeded with further dissection to reveal the empty capsule to confirm the diagnosis. The procedure had consisted of excision of the empty capsule attached to the intended scleral insertion, advancement and reattachment of the retracted "slipped" muscle within its capsule to the sclera. In addition, the ipsilateral lateral rectus was recessed in 4 cases. RESULTS: All patients had presented postoperatively with consecutive exotropia. The tendon step test was positive in all cases. The final results of corrective surgery were "satisfactory" in eight patients (deviation +/ 10 pd or less), "fair" in two (+/115 pd or less) and "unsatisfactory" in 1 (+/-15 or more). CONCLUSION: The tendon step test described herein along with the identification of the empty capsule are valuable adjuncts to the diagnosis of "slipped" muscle. PMID- 11874381 TI - Grand rounds #65: a case of a left hypertropia which decreases markedly in both upgaze and downgaze. PMID- 11874382 TI - Spontaneous intracranial hypotension with unique strabismus due to third and fourth cranial neuropathies. AB - PURPOSE: To report an atypical case of Spontaneous Intracranial Hypotension (SIH) with bilateral trochlear nerve palsies and an oculomotor nerve palsy. DESIGN & METHOD: Case report and literature review. RESULTS (CASE REPORT): A 42 year old man was treated for SIH. His neurological symptoms resolved following neurosurgical treatment with the exception of diplopia due to bilateral trochlear nerve palsies and a pupil-sparing oculomotor nerve palsy. The cranial nerve palsies are believed to be secondary to brainstem ischemia and compression occurring during the acute phase of events. They did not spontaneously improve, but were treated successfully with eye muscle surgery. CONCLUSION: SIH is a rare disease that has been associated with a variety of symptoms and signs including cranial neuropathies. A diagnosis of SIH should be considered in a patient presenting with headache, diplopia secondary to cranial neuropathy and typical radiologic features. This is the first reported case in which bilateral trochlear nerve paresis has been reported in association with this condition. PMID- 11874383 TI - Clinical update on sildenafil citrate. PMID- 11874384 TI - Maximizing the value of medicines by including pharmacogenetic research in drug development and surveillance. AB - Genetics provides significant opportunities to maximize the safety and efficacy of medicines. Over the next 3--5 years, it may be possible to develop tools that use selective information from patients' DNA to enable healthcare professionals to predict more accurately those patients at risk of serious adverse events to some medicines currently available. This is likely to be followed, over the next 5--10 years, by the application of the technology to predict more accurately if individual patients will obtain a therapeutic benefit from a particular medicine. The ability to accurately predict patient response will inevitably change the way medicines are developed, evaluated, and prescribed. Advances in single nucleotide polymorphism (SNP) map technology are likely to drive this innovation. Abbreviated SNP profiles will provide the means to define medicine response tests, thereby allowing clinicians to select the medicine to which the patient is likely to gain the greatest benefit and least risk. This will help to maximize efficacy and reduce the incidence of drug-related adverse events. It may be possible to identify SNP profiles during larger Phase II clinical trials which predict efficacy, and use these to form the basis of Phase III entry criteria. As a result, Phase III trials may be streamlined for many medicines making them smaller, more efficient, and more focused. In addition it may be possible to incorporate pharmacogenetics into postmarketing surveillance strategies to provide a means to identify SNPs which predict uncommon serious adverse drug reactions, and so refine the initial medicine response test. The ability to develop drugs with a predictable response will allow clinicians to provide targeted treatment for patients, with greater confidence of safety and efficacy. Patients therefore will receive more efficacious, timely, and well-tolerated medicines. The challenge for those involved in drug development is to model and evaluate the application of pharmacogenetics so that steps can be taken to realize this potential. PMID- 11874385 TI - Pharmacokinetics of moxifloxacin, a novel 8-methoxy-quinolone, in patients with renal dysfunction. AB - AIMS: To evaluate the influence of impaired renal function on the plasma and urinary pharmacokinetics of moxifloxacin, a novel 8-methoxy-quinolone antibacterial drug. METHODS: Twenty male and 12 female subjects (8 healthy subjects, 24 patients with impaired renal function), 18--75 years of age were investigated in parallel fashion with four groups stratified according to creatinine clearance (CLCR; n=8 for each group). The pharmacokinetics of moxifloxacin and the metabolites M1 (sulphonate metabolite) and M2 (glucuronide) in plasma and urine were determined repeatedly up to 96 h after single oral doses of 400 mg. Patients were monitored intensively with regard to clinical and laboratory safety and tolerability. RESULTS: Single doses of 400 mg moxifloxacin were safe and well tolerated. The urinary excretion of moxifloxacin (Aeur, P: 0.0002) and renal clearance (CLR, P<0.0001) were reduced with decreasing CLCR, mean Cmax was slightly reduced (Cmax-ratio 85.0%, 90% CI 67.9, 106.4% severe renal impairment vs healthy subjects) but the AUC was unchanged even in severe renal impairment (AUC-ratio 101.3%, 90% CI 79.7, 128.6%). The mean AUC of the N sulphonate M1 was slightly increased (by about 53% for the most severe disease) by impaired renal function, but there was no significant correlation between individual AUC and CLCR, whilst Aeur and CLR were significantly correlated with CLCR. In contrast, for the acylglucuronide M2, Aeur (P: 0.0026), CLR (P<0.0001) and AUC (P: 0.0011) were directly correlated with CLCR. CONCLUSIONS: Renal dysfunction had little effect on the plasma pharmacokinetics of either moxifloxacin or metabolite M1, although their renal clearance and urinary excretion were reduced. In contrast renal dysfunction did result in changes in the plasma pharmacokinetics of metabolite M2, causing greater and longer exposure. However the extent of these changes is unlikely to be of clinical relevance. PMID- 11874386 TI - Sex and age differences in the pharmacokinetics of alosetron. AB - AIMS: To determine the effects of sex and age on the pharmacokinetics of alosetron. METHODS: Single oral and intravenous 2 mg doses of alosetron were administered on separate occasions to 48 healthy, young and elderly, males and females. Serum was sampled for 12 h post-dose to measure alosetron concentrations. RESULTS: Serum concentrations of alosetron were higher in females than in males, resulting from a sex difference in clearance by metabolism. Mean clearance values were 504 vs 677 ml min(-1) in young females vs males (mean ratio 0.75), and 461 vs 670 ml min(-1) in elderly females vs males (mean ratio 0.69). The sex difference in alosetron pharmacokinetics achieved statistical significance in the elderly, but not in the young. Irrespective of sex, alosetron clearance was increased by smoking. Serum concentrations tended to be higher in the elderly, although the effect of age was generally not significant. Volume of distribution was smaller in females (approximately 63 l) compared with males (approximately 84 l), regardless of age or the sex difference in body weight. CONCLUSIONS: A significant difference in clearance by metabolism of alosetron between the sexes, and possibly between the young and elderly was observed. PMID- 11874387 TI - Efficacy of calcium channel blockers as maintenance therapy for asthma. AB - AIMS: Previous bronchoprovocation studies indicate that nifedipine attenuates airway responsiveness to several stimuli whereas diltiazem has no effect. The aim of this study was to determine whether such studies predict the efficacy of calcium channel blockers as maintenance therapy for persistent asthma. METHODS: Twenty-one otherwise healthy adults with persistent asthma, mean age 25 years, completed treatment with maximum tolerated doses of placebo (P), nifedipine (N), and diltiazem (D) in a double-blind, randomized, three-treatment, three-period, crossover manner, each for 4 weeks. Frequency and severity of asthmatic symptoms were recorded twice daily, as well as peak expiratory flow and frequency of 'prn' use of inhaled terbutaline. Blood pressure, heart rate, P-R interval of the ECG and spirometry were measured biweekly. At the end of each treatment, airway responsiveness to exercise was measured. RESULTS: The mean (s.e. mean)% of days with wheeze was 69plus minus7% during P, 75plus minus6% during N and 72plus minus6% during D (P=0.7). FEV1 was 79plus minus2% of predicted during P, 81plus minus2% during N and 79plus minus2% during D (P=0.6). The decrease in FEV1 after exercise was 32plus minus4% during P, 32plus minus5% during N and 27plus minus4% during D (P=0.5). Heart rate was elevated during N (P=0.0002) whereas P-R interval was prolonged during D (P=0.0001). CONCLUSIONS: Maintenance therapy with calcium channel blockers, at doses that produce cardiovascular effects, do not suppress the signs and symptoms of persistent asthma. Previous bronchoprovocation studies did not predict these results. PMID- 11874388 TI - Comparative activity of cetirizine and mizolastine on histamine-induced skin wheal and flare responses at 24 h. AB - AIMS: The aim of our study was to compare the activity of cetirizine 10 mg with that of mizolastine 10 mg vs placebo at 24 h after intake in healthy volunteers. METHODS: This was a double-blind, randomized, placebo controlled, three-way cross over study with a wash-out period of 7 +/- 2 days between each period. The study included 36 healthy volunteers (18--50 years, mean age = 32 years; 9 males). The objective measurement was the cutaneous reactivity to increasing concentrations of histamine (0, 5, 10, 20, 40, 80, 160 mg ml(-1)) administered by prick tests. The reactivity was evaluated by the wheal and flare areas (mm2). The AUC (area under curves) values of the wheal and flare areas as a function of the log2 transformed histamine concentration were calculated for each subject and treatment, and compared. RESULTS: A highly significant treatment effect was evidenced both for wheal and flare responses (P = 0.0001). This indicates the good activity of both cetirizine 10 mg and mizolastine 10 mg in inhibiting skin wheal and flare reactions to histamine. In addition, the mean AUC values significantly differed between cetirizine and mizolastine (64.8 and 117.8 log2 (mg ml(-1)) x mm2 for wheal, and 939.4 and 2340.8 for flare, respectively; P = 0.0001), with a superior activity of cetirizine than mizolastine at 24 h after intake both on wheal and flare responses. The tolerance of cetirizine and mizolastine was good. The severity of the adverse events was never more than 'moderate', 'fatigue' being the most frequent reported symptom [cetirizine (6 subjects), placebo (3), mizolastine (5)], followed by 'somnolence' [cetirizine (0), placebo (1), mizolastine (3)]. There was no serious adverse event. CONCLUSIONS: This study shows that cetirizine (10 mg) suppresses skin reactivity to histamine more effectively than mizolastine (10 mg) 24 h after intake in healthy volunteers. PMID- 11874389 TI - Limitation of the in vitro whole blood assay for predicting the COX selectivity of NSAIDs in clinical use. AB - AIMS: To assess if the inhibitory potency of nonsteroidal anti-inflammatory drugs (NSAIDs) on cyclooxygenase (COX) isoenzymes, when given therapeutically in humans, can be predicted from their in vitro concentration-response curves using the whole blood assay. METHODS: Twenty-four healthy male volunteers aged 20--27 years were recruited. Inhibition of blood COX isoenzymes was determined in vitro before any drug intake and ex vivo after single and repeated intake of either 7.5 mg meloxicam once, 400 mg ibuprofen three times daily or 75 mg diclofenac SR once, taken in a randomized cross-over design. Production of thromboxane B2 (TXB2) during clotting and of prostaglandin E2 (PGE2) during endotoxin exposure served as indicators of platelet COX-1 and monocyte COX-2 activity, respectively. Drugs were determined in plasma by h.p.l.c., with a chiral separation of ibuprofen and free fractions after equilibrium dialysis. RESULTS: Intra-subject variation for COX-1 and COX-2 at baseline was at 26 +/- 18% and 18 +/- 13% respectively, and intersubject variation at 39% and 36%, respectively. The ratios of IC50s and, at best, of IC80s revealed diclofenac and meloxicam as selective COX-2 inhibitors and ibuprofen as a preferential COX-1 inhibitor in vitro. However, after oral intake, ibuprofen inhibited ex vivo COX-2 by 80% whereas diclofenac inhibited COX-1 by 70%. Meloxicam inhibited COX-1 from 30 to 55% depending on the repetition of the dose and increase in plasma concentrations. Using in vitro dose--response curves, the in vivo inhibitory potency of diclofenac was estimated adequately from its circulating concentration ([-0.18, 0.21] for COX-1 and [-0.13, -0.03] for COX-2) but this was not the case for ibuprofen on COX-2 ([-0.14, 0.27]) and meloxicam on COX-1 ([0.31, 1.05]). The limited predictability of the system was not improved through considering the unbound fraction of the drugs or the variable chiral inversion of ibuprofen. CONCLUSIONS: Assessment of COX-2 selectivity based on in vitro studies and pharmacological modelling has a limited clinical relevance. There is a need to investigate COX selectivity at therapeutic plasma concentrations of NSAIDs using the ex vivo whole blood assay. PMID- 11874390 TI - 5-HT1B-receptors and vascular reactivity in human isolated blood vessels: assessment of the potential craniovascular selectivity of sumatriptan. AB - AIMS: 5-HT1B-receptor mediated vasoconstriction of cranial arteries is a potential mechanism by which 5-HT1B/1D-receptor agonists such as sumatriptan produce their antimigraine effects. 5-HT1B-receptors exist in other blood vessels which may give rise to unwanted vascular effects. Therefore we examined the distribution of 5-HT1B-receptor immunoreactivity (i.r.) in human blood vessels (including target and nontarget vessels) and confirmed the functionality of this receptor protein, by comparing the vasoconstrictor effects of sumatriptan and 5 HT (the endogenous ligand) in isolated vessels. METHODS: Blood vessels (middle meningeal, pial, temporal and uterine arteries and saphenous veins) were obtained from surgical patients (with consent). Sections of the vessels were prepared for routine immunohistochemical studies using specific 5-HT1B- and 5-HT1D-receptor antibodies. For functional studies, ring segments of the vessels were mounted in organ baths for isometric tension recording. RESULTS: 5-HT1B-receptor i.r. was detected on the smooth muscle layer in middle meningeal, pial and uterine arteries and in saphenous vein and sumatriptan produced contractions in these vessels with potency values (mean pEC50) of 7.00, 7.08, 6.44 and 6.61, respectively, the magnitude of contraction was greatest in the cranial arteries with Emax values of 100.7, 60.3, 23.0 and 35.9%, respectively (expressed as a percentage of the reference agonist 45 mm KCl). 5-HT1B-receptor i.r. was not detected in temporal artery and sumatriptan had no effect in this artery. 5-HT1D receptor i.r. was not detected in any of the vessels studied. CONCLUSIONS: Sumatriptan can evoke vasoconstriction in antimigraine target vessels and also in nontarget vessels through an action at 5-HT1B-rcceptors. Sumatriptan acts preferentially to cause contraction in human cranial arteries compared with the other blood vessels we examined and this effect is likely to be shared by other drugs of this class. PMID- 11874392 TI - Levormeloxifene: safety, pharmacodynamics and pharmacokinetics in healthy postmenopausal women following single and multiple doses of a new selective oestrogen receptor modulator. AB - AIMS: The safety, pharmacodynamics and pharmacokinetics of levormeloxifene, a selective oestrogen receptor modulator (SERM), were investigated in postmenopausal women following single doses and multiple dosing once daily up to 56 days. METHODS: The two randomized, double-blind, placebo controlled studies of six single ascending doses and at four multiple dose levels, respectively, included a total of 104 healthy postmenopausal women. Safety assessments comprised vital signs, ECG, haematology, clinical chemistry and reporting of adverse events. The pharmacodynamic properties were investigated after multiple dosing by assessment of the short-term effects on bone and lipid metabolism and on the hypothalamic-pituitary axis. Blood samples for pharmacokinetic analysis were collected at intervals until 648 h (27 days) after single and multiple dosing. RESULTS: Levormeloxifene was tolerated well after single doses in the range of 2.5--320 mg and multiple once daily dosing in the range of 20--160 mg. Adverse events reported were generally mild or moderate. The most frequent adverse events after multiple dosing were headache, abdominal pain and leukorrhea with the highest frequency reported after the highest daily dose of 160 mg levormeloxifene. Five weeks of treatment with 20--160 mg levormeloxifene and 8 weeks of treatment with 40 or 80 mg levormeloxifene reduced the biochemical marker of bone turnover, the collagen I C-terminal telopeptide (CrossLaps) by 44.4% [95% CI: 11.3, 65.1] and 35.5% [95% CI: 14.0, 51.6], respectively, without any dose-dependent decrease in the studied dose range. The total cholesterol and LDL-cholesterol concentrations were significantly reduced by 19--25% and 28--35%, respectively, when compared with placebo. HDL-cholesterol and triglyceride concentrations were not affected. An oestrogen-like effect on the hypothalamic pituitary axis was observed with approximately 50% reductions of FSH and LH after 8 weeks of treatment. No clinically significant changes of other safety variables were observed. The pharmacokinetic analysis demonstrated a rapid absorption (mean tmax: 2--3 h), a slow elimination (mean t1/2: 4.8--8.4 days) and dose linearity of Cmax and AUC for doses up to 160 mg. As expected for a drug with slow elimination given frequently, the relative fluctuation around the steady state plasma concentration was small and the drug accumulation considerable (RA: 3--5). CONCLUSIONS: Short-term administration of levormeloxifene in postmenopausal women was well-tolerated at doses that elicited a favourable pharmacodynamic response suggesting oestrogen-like bone preserving and antiatherogenic effects. Little variation of peak-trough plasma concentrations was observed during daily administration due to a plasma half-life of approximately 1 week. PMID- 11874391 TI - Investigation of the effect of chlormethiazole on cerebral chemistry in neurosurgical patients: a combined study of microdialysis and a neuroprotective agent. AB - AIMS: Promising pre-clinical results from laboratory studies of neuro-protective drugs for the treatment of patients with stroke and head injury have not been translated into benefit during clinical trials. The objective of the study was to assess the feasibility of administrating a potential neuro-protective drug (chlormethiazole) in conjunction with multimodality monitoring (including microdialysis) to patients with severe head injury in order to determine the effect of the agent on surrogate endpoints and penetration into the brain. METHODS: Multimodality monitoring including cerebral and peripheral microdialysis was applied to five head-injured patients on the neuro-intensive care unit. Chlormethiazole (0.8%) was administered as a rapid (10 ml min(-1)) intravenous loading infusion for 5 min followed by a slow (1 ml min(-1)) continuous infusion for 60 min. The following parameters were monitored: heart rate, mean arterial blood pressure, intracranial pressure, cerebral perfusion pressure, peripheral oxygen saturation, continuous arterial oxygen partial pressure, arterial carbon dioxide partial pressure, arterial pH, arterial temperature, cerebral tissue oxygen pressure, cerebral tissue carbon dioxide pressure, cerebral pH, cerebral temperature, electroencephalograph (EEG), bi-spectral index, plasma glucose, plasma chlormethiazole, and cerebral and peripheral microdialysis assay for chlormethiazole, glucose, lactate, pyruvate and amino acids. RESULTS: Despite achieving adequate plasma concentrations, chlormethiazole was not detected in the peripheral or cerebral microdialysis samples. The drug was well tolerated and did not induce hypotension, hyperglycaemia or withdrawal seizures. The drug did not change the values of the physiological or chemical parameters including levels of GABA, lactate/pyruvate ratio and glutamate. The drug did, however, induce EEG changes, including burst suppression in two patients. CONCLUSIONS: Chlormethiazole can be safely given to ventilated patients with severe head injury. There was no evidence of hypotension or withdrawal seizures. Combining a pilot clinical study of a neuro-protective agent with multimodality monitoring is feasible and, despite the lack of effect on physiological and chemical parameters in this study, may be a useful adjunct to the development of neuro-protective drugs in the future. Further investigation of the capability of microdialysis in this setting is required. By investigating the effect of a drug on surrogate end points, it may be possible to identify promising agents from small pilot clinical studies before embarking on large phase III clinical trials. PMID- 11874393 TI - Differential cognitive effects of ebastine and (+)-chlorpheniramine in healthy subjects: correlation between cognitive impairment and plasma drug concentration. AB - AIMS: It has been widely recognized that classical antihistamines induce sedation as an adverse effect, while second-generation antihistamines have few if any sedative effects. In order to evaluate the sedative properties of ebastine, a second-generation antihistamine, its effect on cognitive performance in healthy subjects was compared with placebo and (+)-chlorpheniramine. METHODS: Twelve healthy male subjects were instructed to perform six types of attention-demanding cognitive tasks, and objective measurements of reaction times and accuracy was made before and after drug administration. Their sleepiness levels were also monitored. Test drugs were ebastine 10 mg, placebo and two doses of (+) chlorpheniramine 2 mg and 6 mg, as positive controls. Plasma drug concentrations at the end of the study were analysed. RESULTS: After treatments with (+) chlorpheniramine, the reaction times of the tasks were significantly prolonged (e.g. ratios of after/before dosing: placebo (0.998 +/- 0.113) vs (+) chlorpheniramine 2 mg (1.103 +/- 0.083; P<0.05) or (+)-chlorpheniramine 6 mg (1.170 +/- 0.139; P<0.001) in a 7 ms visual discrimination time task) and the accuracy was significantly decreased (e.g. ratios: placebo (1.038 +/- 0.158) vs (+)-chlorpheniramine 2 mg (0.792 +/- 0.202; P<0.01) or (+)-chlorpheniramine 6 mg (0.837 +/- 0.222; P<0.05) in a 7 ms task). On the other hand, performance was not affected by ebastine or placebo treatment (e.g. ebastine 10 mg (reaction time ratio; 1.014 +/- 0.067 and accuracy ratio; 0.990 +/- 0.146) in a 7 ms task). Subjective sleepiness was also not affected by ebastine but (+)-chlorpheniramine significantly increased sedation. With respect to the relationship between plasma drug concentrations and task performance, the latter deteriorated with an increase in plasma (+)-chlorpheniramine concentration (e.g. r=0.439 (P=0.007) in a 5 ms and r = 0.352 (P=0.039) in a 7 ms task), but it did not correlate with the plasma concentration of carebastine, an active metabolite of ebastine. CONCLUSIONS: Ebastine 10 mg did not cause any cognitive impairment or subjective sleepiness. On the other hand, (+)-chlorpheniramine impaired cognitive function and induced sleepiness even at 2 mg, the recommended dose in over-the-counter medication. In addition, impaired CNS performance was significantly correlated with plasma (+)-chlorpheniramine concentration. PMID- 11874394 TI - Early clinical experience with the novel NMDA receptor antagonist CNS 5161. AB - AIMS: To investigate the safety, tolerability and pharmacokinetics of the novel NMDA antagonist CNS 5161 in humans. Excessive activation of glutamate receptors, especially of the N-methyl-d-aspartate (NMDA) subtype has been associated with neuropathic pain, and brain damage caused by focal ischaemia in mature brain or hypoxia-ischaemia (HI) in neonates. CNS 5161 is a novel NMDA ion-channel antagonist that interacts with the NMDA receptor/ion channel site to produce a noncompetitive blockade of the actions of glutamate. Pre-clinical studies have demonstrated neuroprotective effects of CNS 5161 in the adult rat model of focal cerebral ischaemia, as well as anticonvulsant and analgesic effects. This study reports the first administration of CNS 5161 to man. Its objectives were to investigate the haemodynamic effects of the compound, to assess its safety and tolerability in healthy male volunteers, and to provide some preliminary human pharmacokinetic data. METHODS: We performed a randomized, double-blind placebo controlled phase 1 dose escalation study of CNS 5161. Volunteers were randomized to receive CNS 5161 or placebo in a ratio of 3:1. Twenty-four of 32 healthy volunteers received intravenous infusion of CNS 5161 over 15 min, followed by serial measurements of plasma drug concentration and haemodynamic observations over 24 h. A dose escalation design was adopted and the volunteers were stratified into eight dosage groups, ranging from 30 microg to 2000 microg. RESULTS: The drug was well tolerated by recipients. Side-effects were dose related, self limiting and comprised minor subjective sensory symptoms. A dose dependent rise in systolic, mean arterial and diastolic blood pressure was seen in subsequent dosage groups, reaching 23/19 mmHg. Maximal effects were seen between 60 and 120 min after commencement of infusion. All subjects returned to baseline haemodynamic values within 24 h. Putative neuroprotective concentrations of CNS 5161 were achieved transiently, although these levels were not sustained. The pharmacokinetic data were best described by a two compartment model. The mean half-life was 2.95 h (s.d. 0.75). Mean clearance was 106 l h(-1) (s.d. 17.8) mean volume of distribution was 296 l (s.d. 69). These parameters were not significantly affected by body weight. CONCLUSIONS: This study suggests that CNS 5161 is well tolerated in healthy volunteers within the dose range studied. In addition, information concerning the pharmacokinetics of the compound has been acquired. Studies to investigate the efficacy of the compound in man may now be justified. PMID- 11874395 TI - Activated charcoal alone or after gastric lavage: a simulated large paracetamol intoxication. AB - AIMS: Activated charcoal is now being recommended for patients who have ingested potentially toxic amounts of a poison, where the ingested substance adsorbs to charcoal. Combination therapy with gastric lavage and activated charcoal is widely used, although clinical studies to date have not provided evidence of additional efficacy compared with the use of activated charcoal alone. There are also doubts regarding the efficacy of activated charcoal, when administered more than 1 h after the overdose. The aim of this study was to examine if there was a difference in the effect of the two interventions 1 h post ingestion, and to determine if activated charcoal was effective in reducing the systemic absorption of a drug, when administered 2 h post ingestion. METHODS: We performed a four limbed randomized cross-over study in 12 volunteers, who 1 h after a standard meal ingested paracetamol 50 mg kg(-1) in 125 mg tablets to mimic real-life, where several factors, such as food, interfere with gastric emptying and thus treatment. The interventions were activated charcoal after 1 h, combination therapy of gastric lavage followed by activated charcoal after 1 h, or activated charcoal after 2 h. Serum paracetamol concentrations were determined by h.p.l.c. Percentage reductions in the area under the curve (AUC) were used to estimate the efficacy of each intervention (paired observations). RESULTS: There was a significant (P<0.005) reduction in the paracetamol AUC with activated charcoal at 1 h (median reduction 66%, 95% confidence intervals 49, 76) compared with controls, and a significant (P<0.01) reduction for gastric lavage followed by activated charcoal at 1 h (median reduction 48.2%, 95% confidence interval 32.4, 63.7) compared with controls. There was no significant difference between the two interventions (95% confidence interval for the difference -3.8, 34.0). Furthermore, we found a significant (P<0.01) reduction in the paracetamol AUC when activated charcoal was administered 2 h after tablet ingestion when compared with controls (median 22.7%, 95% confidence intervals 13.6--34.4). CONCLUSIONS: These results suggest that combination treatment may be no better than activated charcoal alone in patients presenting early after large overdoses. The effect of activated charcoal given 2 h post ingestion is substantially less than at 1 h, emphasizing the importance of early intervention. PMID- 11874396 TI - Patient reporting of potential adverse drug reactions: a methodological study. AB - AIMS: To develop a systematic generic method of enabling patients to report symptoms which they believe to be due to a particular prescribed drug. METHODS: A piloted body system-based questionnaire was distributed to patients registered with 79 medical practices in Grampian prescribed one of nine recently marketed 'black triangle' drugs. These comprised four antidepressants, three antiepileptics and two analgesics. This requested respondents to identify any symptoms experienced over the previous year which they thought could be due to the 'black triangle' drug they had used. A sample of medical records was examined to compare symptoms recorded with those reported by patients. A classification system was developed for the study to enable the assessment of symptoms reported for their potential relationship to patients' drug therapy. All symptoms reported were classified, taking into account information provided by patients on their concomitant drugs and diseases. A specialist pharmacist independently re classified a sample of the symptoms to validate the process. RESULTS: A 36.3% response rate was obtained (837/2307) with 742 respondents (88.6%) reporting at least one symptom. The median per patient was 6.0 (range 0--71), with almost half (406, 48.5%) reporting fewer than five symptoms. Most symptoms (71.0%) were classified as being probably or possibly related to the drugs studied. Agreement between researcher and specialist on the classification of 75.3% of 716 symptoms was obtained (Kappa=0.563). Responses from patients prescribed antidepressant drugs were more likely to include symptoms potentially caused by these drugs (74.5% of all symptoms reported) than those from patients prescribed analgesics (67.4%) or antiepileptics (65.1%, chi2 = 23.858, d.f. = 2, P < 0.001). Patients reporting large numbers of symptoms were more likely to report some which were classed as unlikely to be an ADR or unattributable (chi2 = 80.587, d.f. = 3, P < 0.001). Of the 742 reporting symptoms in questionnaires, 402 (54.2%) claimed to have reported some or all of these to their doctor. Only 162 (22.6%) of 716 patient-reported symptoms were documented in the primary care medical records of 103 patients prescribed tramadol or venlafaxine. CONCLUSIONS: Respondents were clearly willing to report symptoms, the majority of which were classed as possibly/probably related to the drugs studied. The results suggest that patients do not report all symptoms they suspect to be ADRs to their GP and that GPs do not record all symptoms which may be reported to them. The method could help to identify problems which patients perceive as being related to their drug therapy and contribute to increased ADR reporting. PMID- 11874397 TI - Hospital prescribing errors: epidemiological assessment of predictors. AB - AIMS: To demonstrate an epidemiological method to assess predictors of prescribing errors. METHODS: A retrospective case-control study, comparing prescriptions with and without errors. RESULTS: Only prescriber and drug characteristics were associated with errors. Prescriber characteristics were medical specialty (e.g. orthopaedics: OR: 3.4, 95% CI 2.1, 5.4) and prescriber status (e.g. verbal orders transcribed by nursing staff: OR: 2.5, 95% CI 1.8, 3.6). Drug characteristics were dosage form (e.g. inhalation devices: OR: 4.1, 95% CI 2.6, 6.6), therapeutic area (e.g. gastrointestinal tract: OR: 1.7, 95% CI 1.2, 2.4) and continuation of preadmission treatment (Yes: OR: 1.7, 95% CI 1.3, 2.3). CONCLUSIONS: Other hospitals could use our epidemiological framework to identify their own error predictors. Our findings suggest a focus on specific prescribers, dosage forms and therapeutic areas. We also found that prescriptions originating from general practitioners involved errors and therefore, these should be checked when patients are hospitalized. PMID- 11874398 TI - Flow-mediated nitric oxide activity in the forearm vasculature of premenopausal women. AB - AIMS: Nitric oxide (NO) is involved in acute flow-mediated vasodilatation in various vascular beds. We determined whether acutely increasing flow in the human forearm of premenopausal women increases vascular NO activity. METHODS: Forearm blood flow (FBF) was measured by venous occlusion plethysmography. Responses to brachial artery infusion of noradrenaline (a control vasoconstrictor, 20, 50, and 100 ng min(-1), each for 5 min) and NG-monomethyl l-arginine (L-NMMA), an NO synthase inhibitor (200, 400, and 800 microg min(-1), each for 5 min), were determined in eight premenopausal women before and following elevation of basal FBF with glyceryl trinitrate (GTN) on two separate occasions. RESULTS: Flow elevation with GTN increased responses to L-NMMA (summary measure 103 +/- 12 vs 65 +/- 12 arbitrary units, P<0.05), but not to noradrenaline (95 +/- 35 vs 74 +/- 12, P=0.50). CONCLUSIONS: Acute elevation of FBF in nonpregnant women is associated with enhanced responses to NO synthase inhibition, consistent with flow-mediated increased NO activity. PMID- 11874399 TI - The influence of body weight on the pharmacokinetics of mefloquine. PMID- 11874400 TI - Paracetamol can exacerbate irradiation-induced DNA damage. PMID- 11874402 TI - Management and outcome of central precocious puberty. PMID- 11874403 TI - Assessment of selective arterial calcium stimulating and hepatic venous sampling to localize insulin-secreting tumours. PMID- 11874404 TI - Hyperkalaemia and selective hypoaldosteronism in myotonic dystrophy. PMID- 11874405 TI - Severe impairment of bone mass and turnover in Cushing's disease: comparison between childhood-onset and adulthood-onset disease. AB - BACKGROUND: Osteoporosis is an important, frequently unrecognized consequence of hypercortisolism. OBJECTIVE: To evaluate whether the age of onset of hypercortisolism influences its effects on bone mass and turnover. SUBJECTS: 10 with childhood-onset (co) and 18 with adulthood-onset (ao) Cushing's disease (CD); 28 age-, sex- and body mass index (BMI)-matched healthy subjects served as controls. STUDY DESIGN: Open, cross-sectional controlled. MEASUREMENTS: Bone mineral density (BMD) at lumbar spine, serum osteocalcin (OC), and urinary N telopeptides of type I collagen (Ntx) levels. RESULTS: BMD at lumbar spine was significantly lower in all CD patients than in controls (Z score, -2.3 +/- 0.1 vs. -0.2 +/- 0.01; P < 0.001). co-CD and ao-CD patients had similar values of bone mass when expressed as Z score (-2.6 +/- 0.4 vs. -2.1 +/- 0.2; P = 0.27) or as BMD (0.728 +/- 0.03 vs. 0.78 +/- 0.03 g/cm2; P = 0.25). In particular, osteoporosis was observed in 16 patients (57.1%) [eight adolescents (80%) and eight adults (44.4%)] and none of the controls; osteopenia was found in two co-CD patients (20%) and none of the healthy adolescents, 10 ao-CD patients (55.6%) and four healthy adults (14.3%) (chi2 = 7.87, P < 0.01; chi2 = 2.99, P = 0.09, respectively). In co-CD and ao-CD patients, serum OC levels were similar and significantly lower than in controls (P < 0.01); urinary Ntx levels were significantly higher than in controls (P < 0.001) and were significantly higher in co-CD than in ao-CD patients (P < 0.001). No significant correlation was found between urinary cortisol levels, serum cortisol and age and lumbar Z score values, while a significant correlation was found between Ntx levels and disease duration (r = 0.434; P = 0.021) and plasma cortisol (r = 0.440; P = 0.019). CONCLUSIONS: Cushing's disease causes bone loss and abnormalities of bone turnover both in childhood-onset and in adulthood-onset patients. A strict follow up of bone mass and turnover is mandatory in all patients with Cushing's disease to prevent fractures later in life and specific treatment for bone loss is strongly suggested. PMID- 11874406 TI - Fracture risk is increased in patients with GH deficiency or untreated prolactinomas--a case-control study. AB - OBJECTIVE: The pituitary secretes many hormones of significance to bone turnover and thus skeletal integrity. The aim of this study was to examine fracture risk in patients with pituitary disorders with special reference to GH deficiency and hyperprolactinaemia. DESIGN: Case-control study. MEASUREMENTS: Fracture occurrence. PATIENTS: A self-administered questionnaire was issued to 537 consecutive patients with pituitary disorders excluding Cushing's disease. A total of 426 (79%) returned the questionnaire and 422 of these could be analysed. Each respondent was compared to three age- and gender-matched control respondents to the same questionnaire drawn randomly from the background population. RESULTS: The patients had a mean age of 51.4 +/- 14.8 years. One hundred and eight patients had acromegaly, 86 had prolactinomas, 136 had non-functioning pituitary adenomas (NFPA), 23 had craniopharyngiomas, and 73 had other types of pituitary disorders. For the total group the fracture risk was not elevated either before or after confirmed diagnosis compared to controls. However, among the patients with prolactinomas, the fracture risk was significantly increased before (relative risk, RR = 1.6, 95% CI: 1.1--2.3) but not after diagnosis. In patients with NFPA, fracture risk was borderline significantly elevated following diagnosis (RR = 1.6, 95% CI: 1.0--2.6). Patients with subnormal stimulated peak GH values suggestive of GH deficiency had a significantly higher risk of fractures after diagnosis than patients who had normal stimulated peak GH values (odds ratio, OR = 4.90, 95% CI: 1.10--21.88). CONCLUSIONS: Untreated prolactinomas were associated with a significant increase in fracture risk. Growth hormone deficiency was also associated with a higher fracture risk. PMID- 11874407 TI - Spinal irradiation impairs the osteo-anabolic effects of low-dose GH replacement in adults with childhood-onset GH deficiency. AB - BACKGROUND AND OBJECTIVE: Both adult- and childhood-onset GH-deficient adults are prone to osteopenia. Studies of GH replacement have, for the most part, demonstrated increases in bone mineral density (BMD). Previous studies have, however, used GH doses in excess of those currently used in low-dose titration regimens aimed at normalizing the serum IGF-I level. Furthermore, the effect of GH on the lumbar spine that has been irradiated during treatment of childhood cancer is unknown. PATIENTS: Thirty-two adult patients with childhood-onset GH deficiency were subdivided according to whether or not they had received spinal irradiation in childhood. The cohort in whom the spine had not been irradiated was comprised of 17 patients (seven male, 10 female), median age 29.8 years (range 20.6--40.8), the median age at primary pathological diagnosis being 9 years (range 4--16). The cohort who received spinal irradiation was composed of 15 patients (seven male, eight female), median age 22.9 years (range 16.5--40.3), with a median age at craniospinal irradiation of 9 years (range 2--16). MEASUREMENTS: At baseline, BMD was assessed at the lumbar spine and femoral neck by DXA, and at the ultradistal and distal radius by SXA. The patients were then commenced on GH replacement, titrating the dose at 4--6-weekly intervals to normalize the serum IGF-I level. BMD scans were reassessed following at least 1 year of GH replacement therapy. The mean duration of GH therapy was 1.68 plus minus 0.52 years. RESULTS: BMD was significantly reduced, compared to the reference data, at all four sites measured in both the spinally irradiated and unirradiated groups. No significant difference was observed between the subgroups with respect to lumbar spine BMD (P = 0.64). In the cohort who did not receive spinal irradiation, an increase in BMD of 3.5% above baseline was observed at the lumbar spine (P = 0.018), 13/17 patients showing a positive increment in lumbar spine BMD. No significant changes in BMD were observed at any other site within this cohort, or at any site in the patients who received spinal irradiation. A significantly greater change in lumbar spine BMD was observed in the unirradiated spine cohort compared with the spinally irradiated cohort (P = 0.018). No differences in response were demonstrated at the other sites studied between the two subgroups. CONCLUSIONS: We have demonstrated an increase in lumbar spine BMD with long-term (mean 1.78 +/- 0.55 years) GH therapy in adults with childhood onset GH deficiency, who have not received spinal irradiation, when GH was administered by a titration regimen aimed at normalizing the serum IGF-I level. No improvements were observed at the femoral neck, ultradistal or distal radius. Patients who had received spinal irradiation during childhood did not present with a reduction in spinal BMD in excess of that observed in the nonirradiated cohort. The spinally irradiated group were, however, resistant to the osteo anabolic effects of GH, which we propose reflects the capacity of radiation induced damage to suppress the skeletal response. PMID- 11874408 TI - Elderly people with hypothalamic-pituitary disease and untreated GH deficiency: clinical outcome, body composition, lipid profiles and quality of life after 2 years compared to controls. AB - OBJECTIVE: Elderly patients with GH deficiency (GHD) have significant impairments in multiple aspects of quality of life (QOL) but similar lipid profiles compared to age-matched control subjects. There are, however, no data on changes in these parameters with time. This study assessed the impact of untreated GHD over a period of 2 years in a group of elderly patients with hypothalamic-pituitary disease in relation to new illnesses and differences in body composition, circulating lipid profile levels and QOL. Control subjects were also followed for 2 years. SUBJECTS: Twenty-seven elderly patients (> 65 years) with hypothalamic pituitary disorders and GHD (mean peak stimulated GH response 1.6 mIU/l, range 0.6--5.0) were studied initially. Two years later 21 (13 males) agreed to attend for reassessment. Mean age was then 72.7 +/- 5.04 years (range 67--85). Eighteen patients had pituitary tumours, three had craniopharyngiomas. Twenty-seven control subjects were studied at baseline and 17 (7 males) agreed to attend for reassessment. Mean age was then 75.9 +/- 6.97 years (range 67--88). METHODS: Weight, body mass index (BMI), total fat mass (FM) (bioelectrical impedance), serum IGF-1 and fasting lipid profile (total cholesterol, triglyceride, HDL cholesterol, LDL cholesterol) were measured. QOL was assessed in both groups using five interviewer-administered self-rating questionnaires: the Nottingham Health Profile, Short Form-36, Hospital Anxiety and Depression Scale, Mental Fatigue Questionnaire and Life Fulfillment Scale. The GHD group also completed the Disease Impact Scale. RESULTS: Two of the 27 patients with GHD died during the 2-year follow-up (myocardial infarction and probable cerebrovascular accident). Four controls could not be traced but there were no deaths in the other 23. In the 21 GHD patients after 2 years, mean serum IGF-1 and BMI were unchanged (12.6 +/- 5.8 vs. 13.3 +/- 5.1 nmol/l, P = 0.5 and 28.3 +/- 4.3 vs. 29.1 +/- 4.2, P = 0.5, respectively) at the 2-year follow-up and there were no significant changes in the lipid profiles. However, there was a significant reduction in fat mass (31.7 +/- 11.2 vs. 28.5 +/- 10.9%, P = 0.04). In the 17 control subjects after 2 years, serum IGF-1 levels (17.2 +/- 4.0 vs. 15.7 +/- 5.6 nmol/l, P = 0.4), BMI and fat mass were unchanged. However, there was a significant fall in total cholesterol levels over the 2-year follow-up (6.3 +/- 0.9 vs. 5.7 +/- 0.9 mmol/l, P < 0.0001), although LDL cholesterol, triglycerides and HDL cholesterol were unchanged. Analysing the QOL data, the GHD patients had less energy (P < 0.05), more depression (P < 0.05), more pain (P < 0.05) and lower life fulfillment scores (P < 0.01) after 2 years. However, the control subjects also had less energy (P < 0.05), less vitality (P < 0.05) and lower self esteem (P < 0.05), more depression (P < 0.05), worse mental health (P < 0.05), life fulfillment personal (P < 0.01), life fulfillment material (P < 0.02), physical functioning and role physical functioning (P < 0.05) after 2 years. Comparing the patients and controls at baseline, there were significant differences in IGF-1, BMI, FM, LDL cholesterol, personal life fulfillment, mental fatigue, general health and mental health. However, after 2 years, only BMI and depression scores were significantly different. CONCLUSION: These patients with untreated GHD did not have deterioration of body composition or lipid profiles when reassessed after a period of 2 years. In fact, fat mass fell. The control subjects did have a significant decrease in total cholesterol but no change in other lipids or body composition. Some quality of life domains did deteriorate in the patients with GHD. However, the control subjects also had worse quality of life scores after 2 years which were then little different from the GHD patients. These results raise doubts about the benefits of GH replacement in elderly people with GHD. PMID- 11874409 TI - High incidence of mental disorders, reduced mental well-being and cognitive function in hypopituitary women with GH deficiency treated for pituitary disease. AB - OBJECTIVE: Previous studies have shown possible neuroendocrine effects of GH. In the present study we investigated the incidence of mental disorders and the prevalence of mental distress and cognitive dysfunction in hypopituitary women with untreated GH deficiency compared to population-based controls. DESIGN AND PATIENTS: Thirty-three hypopituitary women with a median age of 64 years (range 39--77 years) were investigated cross-sectionally, without any change in hormone substitutions. Twenty-nine of the patients had been operated for a pituitary tumour, 25 had received radiotherapy and 15 had visual dysfunction. The patients were with a very high probability GH deficient, as 29 had subnormal IGF-I levels and the other four were GH deficient as assessed by an insulin tolerance test. The patients were compared with 33 controls matched for sex, age, smoking habits, educational level and residence. MEASUREMENTS: The incidence of mental disorders was calculated from the date of diagnosed hypopituitarism to the time of the present investigation. Mental well-being was assessed by three self-rating questionnaires: the Symptom Checklist-90 (SCL-90), the Interview Schedule for Social Interaction (ISSI) and the social network concept. The subjects were examined with neuropsychological tests of vocabulary (SRB:1 vocabulary test), perceptual speed (WAIS-R Digit Symbol), spatial ability (WAIS-R Block Design), verbal memory (Cronholm--Molander verbal memory test), spatial learning (Austin Maze Test) and reaction time (APT Two-way Reaction Time and APT Inhibition). RESULTS: The hypopituitary women had a higher incidence of mental disorders than the controls; Incidence Rate Ratio 4.5 (95% CI 1.0--21). The Global Severity Index, i.e. the average score of all 90 questions of the SCL-90, was higher in patients (P = 0.001), and the patients had significantly more symptoms of somatization, anxiety, depression, obsession--compulsion, hostility- irritability, phobic and psychotic symptoms (all P less-than-or-equal 0.04). Moreover, 14 patients compared to four controls were classified as possible cases of mental distress according to the SCL-90 (P = 0.006). The patients experienced lower availability of both social attachment (P = 0.02) and integration (P = 0.001), but there were no group differences in the adequacy of these dimensions or in emotional support. The patients had lower scores in four of seven neuropsychological tests (all P less-than-or-equal 0.04). CONCLUSIONS: The hypopituitary women had a higher incidence of mental disorders, more symptoms of mental distress and increased prevalence of cognitive dysfunction. The impaired results in the patients could possibly be explained by several factors, such as transfrontal surgery, radiotherapy, visual dysfunction and unphysiological hormone substitution. Moreover, it is probable that GH deficiency contributed, but placebo-controlled double-blind studies are warranted to investigate whether the psychological dysfunction is reversible on GH substitution. PMID- 11874410 TI - Effects of low-dose oral hydrocortisone replacement versus short-term reproduction of physiological serum cortisol concentrations on insulin action in adult-onset hypopituitarism. AB - OBJECTIVE: Hypercortisolism is associated with impaired glucose tolerance and insulin resistance. For many years hydrocortisone 30 mg was the standard total daily replacement dose in adult hypopituitarism. The use of this conventional dose has now been shown to result in mild biochemical hypercortisolism and might contribute to the increased cardiovascular risk reported in hypopituitarism. The use of lower doses of hydrocortisone replacement therapy might prevent some of the adverse metabolic effects seen with conventional doses. PATIENTS: In a randomized crossover study we assessed peripheral and hepatic insulin action in 15 ACTH-deficient patients with normal glucose tolerance on two occasions while receiving either a low-dose oral hydrocortisone replacement (LOR) therapy (15 mg at 0800, 5 mg at 1700) or a physiological hydrocortisone infusion (PHI), which achieved physiological serum cortisol concentrations. RESULTS: Exogenous glucose infusion rates required to maintain euglycaemia were similar for the LOR and the PHI protocols (26.2 +/- 0.4 vs. 23.8 +/- 0.6 micromol/kg/min, respectively). Endogenous glucose production was also similar (12.0 +/- 2.5 vs. 11.6 +/- 2.4 micromol/kg/min, respectively) and in the post-absorptive state suppressed to a similar extent following insulin (4.5 +/- 2.0 vs. 5.1 +/- 3.1 micromol/kg/min). CONCLUSION: Hydrocortisone replacement therapy at a dose of 15 mg with breakfast, 5 mg with evening meal does not increase peripheral or hepatic insulin resistance when compared to a hydrocortisone infusion designed to simulate physiological serum cortisol concentrations. PMID- 11874412 TI - The corticotrophin-releasing hormone test in preterm infants. AB - OBJECTIVE: The developing hypothalamic--pituitary--adrenal axis (HPAA) may be immature and not yet fully functional in preterm infants. This may result in an inappropriate adrenal response to stress. Little is known about the pituitary- adrenal response to corticotrophin-releasing hormone (CRH) stimulation during the early neonatal period in preterm infants born before 32 weeks of gestation. Therefore, in a first study we investigated the pituitary--adrenal response to 1 microg/kg CRH i.v. in 13 preterm infants born less-than-or-equal 32 weeks of gestation. In addition, in a randomized placebo-controlled study we compared the pituitary--adrenal response of 1 microg/kg CRH to placebo and stimulation with 2 microg/kg CRH. RESULTS: In the first study, the level of ACTH increased from 6.9 +/-2.1 to 11.6 +/- 5.1 pmol/l (P < 0.01) and cortisol increased from 350 plus minus 115 to 582 +/- 201 nmol/l (P < 0.05). Thirty-eight percent of the studied infants showed a maximal level of ACTH < 9 pmol/l, and 15% showed a maximal level of cortisol < 360 nmol/l. In the randomized study, infants in the 1 microg/kg and in the 2 microg/kg CRH group, but not in the placebo group, showed a significant increase in cortisol and ACTH after stimulation (P < 0.01). Stimulated levels of ACTH and cortisol were significantly higher in the 2 microg/kg group compared with the placebo and the 1 microg/kg group. No differences were found for plasma ACTH and cortisol levels in the 1 microg/kg group compared with the placebo group. Basal levels of cortisol and ACTH obtained from the first and from the randomized study correlated significantly (n = 29; r = 0.42, P < 0.03). In addition, in infants stimulated with 1 microg/kg CRH a lower cortisol response correlated with a longer stay in hospital (n = 13; r = --0.57, P < 0.05). CONCLUSIONS: In this study we show that a 1 microg/kg CRH stimulation test in preterm infants results more often in an inappropriate adrenal response while stimulation with 2 microg/kg CRH gives rise to an appropriate response in all studied infants. Furthermore, stimulation with 2 microg/kg CRH results in higher levels of ACTH and cortisol compared to placebo and 1 microg/kg CRH. We conclude that in preterm infants the ability of the pituitary to respond adequately to CRH stimulation depends on the dose of CRH used and may also be dependent on the maturity of the pituitary--adrenal axis. PMID- 11874411 TI - Weight loss increases circulating levels of ghrelin in human obesity. AB - OBJECTIVE: Ghrelin, a novel endogenous ligand for the GH secretagogue receptor, has been reported to have adipogenic actions and induce weight gain in addition to its GH-releasing properties. Interestingly, recent data indicate that ghrelin is downregulated in human obesity, which is also known to be accompanied by reduced GH levels. PATIENTS AND METHODS: To investigate the influence of weight loss on circulating levels of ghrelin we recruited eight obese women among patients attending a 6-month weight-loss course organized by The Danish Heart Association. We measured body composition including computerized tomography as well as fasting plasma ghrelin concentrations before and after weight loss. RESULTS: Plasma ghrelin concentrations increased by 12% following weight loss (P < 0.01), and the increase in ghrelin levels was positively correlated with the extent of weight loss (r = 0.68, P < 0.05). Exposure to exogenous GH intravenously did not influence fasting ghrelin levels either before or after weight loss. Our data further suggest the existence of hyperghrelinaemia in a single subject with long-standing obesity but no signs of GH excess. CONCLUSIONS: This study provides evidence of a reversible suppression of ghrelin associated with obesity. The feasibility of measuring ghrelin in the circulation provides a new tool for the investigation of the complex hormonal regulation of appetite and energy balance. PMID- 11874413 TI - Inhibin B in boys from birth to adulthood: relationship with age, pubertal stage, FSH and testosterone. AB - OBJECTIVE: Inhibin B in males is produced principally by Sertoli cells under the influence of FSH and is thought to have a role in feedback regulation of FSH. The aims of our study were to investigate how inhibin B changes from birth to late adolescence in boys, to derive reference data and to explore its relation with pubertal stage, FSH and testosterone. DESIGN AND SUBJECTS: Blood samples were collected from (i) 366 boys aged 0--18 years to obtain age-related reference data; (ii) 195 boys who had full pubertal staging; and (iii) a cohort of 15 boys studied longitudinally as they approached and entered early puberty. MEASUREMENTS: Dimeric inhibin B was measured by double antibody enzyme-linked immunosorbent assay (ELISA), FSH by immunoradiometric assay (IRA) and testosterone by an extraction radioimmunoassay. RESULTS: Inhibin B was high in infant boys, decreased gradually to a nadir at 6--10 years of age, then increased rapidly in early adolescence to reach a new plateau at 12--17 years. It was detectable in all samples. Age-related reference ranges and data for calculation of SD scores are presented. In prepubertal boys, inhibin B correlated positively with age (P < 0.001), but not with FSH. Inhibin B increased progressively from pubertal stages G1 to G3 but then decreased slightly at stages G4 to G5 (P less than-or-equal 0.01). At stage G2, inhibin B correlated positively with testosterone (P < 0.01) but not with FSH. From stage G3 onwards, inhibin B correlated inversely with FSH (P < 0.01) but lost its relationship with testosterone. In the cohort of boys studied longitudinally, inhibin B increased progressively prior to pubertal onset and further on entry into early clinical puberty (P < 0.05). Testosterone also increased over this period (P < 0.05) but FSH showed no significant change. CONCLUSIONS: The two peaks of inhibin B during infancy and early puberty appear to reflect the two periods of Sertoli cell proliferation in normal human males. During mid-childhood, a relatively constant amount of inhibin B is secreted constitutively. The early FSH-independent increase in inhibin B that precedes clinical puberty and continues to stage G2 may be stimulated by testosterone or other factors from Leydig cells. The inverse relationship between inhibin B and FSH that subsequently develops from mid puberty onwards is consistent with the establishment of a negative feedback loop at this time. PMID- 11874414 TI - Dimeric inhibins in girls from birth to adulthood: relationship with age, pubertal stage, FSH and oestradiol. AB - OBJECTIVE: Inhibin B is produced by granulosa cells in small antral follicles under the influence of FSH, whilst inhibin A is produced by larger follicles and the corpus luteum. The aims of our study were to investigate how these inhibins change from birth to late adolescence in girls, to derive reference data and to explore their relation with pubertal stage, FSH, oestradiol and each other. STUDY DESIGN AND SUBJECTS: Blood samples were collected from: (a) 345 girls aged 0--18 years to obtain age-related reference data, and (b) 80 pre-menarcheal girls with full pubertal staging, of whom 40 were on GH treatment at the time of sampling. MEASUREMENTS: Dimeric inhibins A and B were measured by double antibody enzyme linked immunosorbent assay (ELISA), FSH by immunoradiometric assay (IRMA) and oestradiol by radioimmunoassay. RESULTS: Median inhibin B was low until age 6 years, slightly higher from 6 to 10 years, then increased from 10 to 12 years to reach a plateau from 12 to 18 years. Inhibin A was usually detectable in girls younger than 3 months but thereafter became undetectable in most samples until after age 10 years, when median levels rose progressively to 14 years, then stabilized from 14 to 18 years. Both inhibins displayed considerable scatter about the median throughout infancy, childhood and adolescence. Girls aged 0--10 years showed a positive correlation between inhibins A and B (P < 0.0001), whereas those aged 14--18 years showed an inverse relationship (P < 0.001), indicating the onset of ovulatory cycles. Age-related reference ranges and data for calculation of SD scores are presented. GH-treated girls at pubertal stage B2 (but not at B1 or B3--5) had higher inhibin B and FSH levels than untreated girls and were excluded from further analysis. Both inhibins A and B increased during puberty (P < 0.0001) and were positively correlated with each other (P < 0.01). Both inhibins were also positively correlated with FSH in pre-pubertal girls (P < 0.05) but not at pubertal stages B3--5. CONCLUSIONS: Although median levels of inhibins A and B remained low until after age 10 years in girls, the increased levels of both inhibins in individual samples, together with their positive relationship with FSH, provide further evidence of sporadic follicular development throughout infancy and childhood under the influence of FSH. The increase in both inhibins during puberty and their changing relationship with FSH are in keeping with the concept of follicular growth being dependent on the duration of FSH elevation above a critical threshold rather than the degree of elevation per se. PMID- 11874415 TI - Complete virilization in congenital adrenal hyperplasia: clinical course, medical management and disease-related complications. AB - OBJECTIVE: In girls with congenital adrenal hyperplasia (CAH), genital ambiguity usually leads to a rapid neonatal diagnosis. Rarely, CAH causes complete virilization and male sex assignment with a delayed diagnosis. After being confronted with very specific problems in two of such patients, we collected data of patients with CAH and complete virilization in a nationwide study to delineate specific problems of these rare patients in order to improve their management. DESIGN AND PATIENTS: Through the German Working Group of Paediatric Endocrinology (Arbeitsgemeinschaft Padiatrische Endokrinologie, APE), questionnaires were sent to all members caring for patients with CAH and complete virilization in their endocrine clinics. Data from 16 patients from 10 paediatric endocrine centres were assessed by questionnaire. RESULTS: The following problems have been encountered. (1) Sex assignment/gender identity: initially all patients had a male sex assignment. Six patients were diagnosed during the first month of life. Five were reassigned to female sex immediately, one at the age of 19 months. Except in one girl demonstrating some tomboyish behaviour, gender role behaviour in these patients did not differ from unaffected girls. Ten patients were diagnosed late at 3.4--7 years of age. In seven patients with a late diagnosis, male sex assignment was maintained; one of them expressed some concerns about living as a male. In three patients late sex reversal was performed, gender identity is very poor in one and new sex assignment is currently under consideration. (2) SURGERY: irrespective of the sex assigned, all patients had between one and three surgical procedures, including clitoris reduction and (repeated) vaginoplasties in patients with female sex assignment. Hysterectomy and ovarectomy were performed in patients with male sex assignment. (3) Short stature: patients with a late diagnosis of CAH had extremely advanced bone ages of +6.3 to +9.5 years, leading to severely reduced final height of 137 to 150 cm in adult patients. Patients tended to follow height percentiles of genetic females. One pubertal patient was suicidal due to short stature. (4) Central precocious puberty (CPP): prolonged exposition to adrenal androgens led to CPP in one patient. He was treated with GnRH analogues until gonadectomy. CONCLUSIONS: Patients with CAH and complete virilization have a high risk of being diagnosed late. There are major problems and uncertainties of the patients' families and the treating physicians concerning gender assignment. Gender identity is disturbed in some patients. In addition, multiple surgical procedures are necessary and short stature as well as central precocious puberty might be important to avoid late sequelae. While some surgical interventions are probably unavoidable, most of these issues could be resolved with an early diagnosis. Thus, especially for these patients, a neonatal screening programme for CAH would be of paramount importance. PMID- 11874416 TI - Classical and nonclassical 21-hydroxylase deficiency: a molecular study of Argentine patients. AB - OBJECTIVE: To characterize the molecular basis of the 21-hydroxylase deficiency in a group of Argentine patients presenting the classical and nonclassical forms of the disease. DESIGN: To analyse the frequency of point mutations in the CYP21 gene by DNA amplification and mutation detection. PATIENTS: Forty-one patients from 36 nonrelated families: 25 nonclassical (NC), 11 salt-wasting (SW) and five simple virilizing (SV). A total of 27 parents and 13 nonaffected siblings were also analysed. MEASUREMENTS: Basal steroid hormones and 17-hydroxyprogesterone levels following adrenal stimulation with adrenocorticotrophic hormone were measured, together with an analysis of 10 point mutations in the CYP21 gene. RESULTS: A total of 83% and 74.4% classical and nonclassical chromosomes, respectively, were characterized. The intron 2 mutation was the most prevalent among classical alleles. In addition, a high frequency for R356W was observed in both groups (13.3 and 6.9%, respectively), while V281L was the most frequent mutation among the nonclassical patients with a frequency of 39.5%. No alleles containing P30L were observed, and one de novo mutation (R356W) was found. A total of 68.3% patients were fully genotyped, and all but one showed no genotype/phenotype discrepancy. Though the cut-off value for post-ACTH 17 hydroxyprogesterone stimulation was 30.25 nmol/l (10.00 microg/l), the lowest value observed in the fully genotyped nonclassical group was 42.35 nmol/l (14.00 microg/l). CONCLUSIONS: The high number of unidentified alleles in the nonclassical group suggests that less frequent mutations, or the presence of new ones, might be the cause of the disease in the Argentine population. Alternatively, the cut-off value in the ACTH-stimulated 17-hydroxyprogesterone test might overestimate the diagnosis of the nonclassical form by including some patients with heterozygous status. PMID- 11874417 TI - The use of recombinant human TSH in the follow-up of differentiated thyroid cancer: experience from a large patient cohort in a single centre. AB - BACKGROUND: Periodic evaluation of serum thyroglobulin (Tg) and whole body 131I imaging (131I-WBS) are essential in the follow-up of differentiated thyroid carcinoma (DTC); both diagnostic modalities require stimulation by high levels of TSH. Administration of recombinant human TSH (rhTSH) is an alternative to the withdrawal of thyroid hormone therapy. OBJECTIVE: The aim of this study was to report our experience in the use of rhTSH for the management of patients with DTC. PATIENTS: One hundred and four patients were enrolled in the study. A dose of 10 U of rhTSH therapy was injected intramuscularly for 2 consecutive days; 24 h after the second dose of rhTSH the patients were administered 4--5 mCi of 131I and, 48 h later, WBS was performed. RESULTS: In all patients, baseline mean serum Tg and TSH levels were 2.4 +/- 1.9 ng/ml and 0.0153 +/- 0.0232 mIU/l, respectively. Basal Tg levels were detectable in 58 out of 104 patients. After rhTSH injection, mean serum TSH levels rose to 122.67 +/- 47.36 mIU/l. Stimulated serum Tg levels increased to greater-than-or-equal 5 ng/ml and the 131I-WBS showed an uptake in 18 patients (17.4%). Among them there were three with bone metastases and one with brain metastases, who reported violent skeletal pain and a severe headache, respectively. These were caused by the growth of tumour mass of metastases induced by rhTSH administration. CONCLUSIONS: The use of rhTSH avoids the debilitating effects of hypothyroidism and its use successfully promotes iodine uptake and increases the sensitivity of serum Tg testing. The risk of causing serious side-effects recommends performing skull magnetic resonance and radionuclide bone scan in cases of suspected brain or skeletal metastases. PMID- 11874418 TI - Normalized dyslipidaemia after parathyroidectomy in mild primary hyperparathyroidism: population-based study over five years. AB - OBJECTIVE: Postmenopausal women are at increased risk of primary hyperparathyroidism (pHPT). Secondary dyslipidaemia in pHPT has attracted little attention, although morbidity and mortality associated with cardiovascular diseases have been reported to be increased in these patients. DESIGN: A population-based screening programme was used to recruit postmenopausal women with mild, asymptomatic pHPT (mean serum calcium 2.57 +/- 0.12 mmol/l) and matched controls. MEASUREMENTS AND PATIENTS: Serum lipids, lipoprotein fractions and influences of treatment for the parathyroid disease were studied in 87 case control pairs (mean age 67 years), 69 of whom completed a 5-year follow-up period. RESULTS: pHPT was characterized by decreased serum high-density lipoprotein (HDL)-cholesterol, increased total triglycerides, very-low-density lipoprotein (VLDL)-triglycerides and VLDL-cholesterol levels and an elevated atherogenic index. The differences were more pronounced in the cases with serum parathyroid hormone levels in the normal range and were inversely correlated to the serum parathyroid hormone level. Parathyroidectomy, with or without additive hormone replacement therapy, normalized the dyslipidaemia. Five-year surveillance of pHPT without treatment was associated with a maintained increase in total triglycerides and the atherogenic index and a decrease in HDL-cholesterol levels. CONCLUSION: Proatherosclerotic dyslipidaemia characterizes mild pHPT and is effectively reversed by parathyroidectomy. As dyslipidaemia might contribute to the increased risk of cardiovascular diseases and death observed in pHPT, the findings favour operative intervention rather than conservative surveillance in mild, asymptomatic pHPT in postmenopausal females. PMID- 11874419 TI - Genetic and clinical characterization of sporadic cystic parathyroid tumours. AB - OBJECTIVE: The hyperparathyroidism--jaw tumour (HPT--JT) syndrome is one of the familial disorders characterized by primary hyperparathyroidism and has been linked to the chromosomal region of 1q32--q21. The parathyroid tumours related to this syndrome have shown loss of wild-type alleles at this locus suggesting that inactivation of a tumour suppressor gene might be responsible for the disease. In the majority of these tumours cysts are a prominent feature. By loss of heterozygosity (LOH) studies, we investigated the region of interest in an attempt to clarify its possible role in a series of cystic sporadic parathyroid adenomas. DESIGN AND SUBJECTS: A total of 30 patients diagnosed with sporadic hyperparathyroidism were included in the study, genotyped with 17 polymorphic microsatellite markers at chromosome 1q, and additional markers from 1p and 11q13 which are commonly involved in sporadic parathyroid tumours. The cystic parathyroid tumours were characterized clinically, and immunohistochemistry against PTH was carried out to confirm the parathyroid origin of the cysts. RESULTS: LOH was found in six of 30 tumours (20%) on 1q, six of 30 tumours (20%) on 1p and five of 30 tumours (17%) on 11q13. We found a significant correlation between allelic alterations and the clinical parameters, tumour weight and PTH. Furthermore, we found a significant difference between tumour weight and PTH in cases of cystic parathyroid tumours compared with unselected sporadic cases. CONCLUSIONS: These results suggest that cystic parathyroid tumours might represent a new subgroup among parathyroid tumours based on the genetic and clinical findings. Loss of heterozygosity at 1q further supports the presence of a tumour suppressor gene at this locus. PMID- 11874422 TI - Tracking diabetes in Albania: a natural experiment on the impact of modernization on health. PMID- 11874420 TI - Hyperkalaemia and selective hypoaldosteronism in myotonic dystrophy. AB - Myotonic dystrophy (MyD) is a common genetic neuromuscular disorder in which chromosome 19 gives rise to an abnormal expansion of CTG-trinucleotide repeats. MyD is a highly variable multisystem disorder with muscular and nonmuscular abnormalities. Increasingly, endocrine abnormalities, such as gonadal, pancreatic, and adrenal dysfunction are being uncovered. Herein we present three unrelated cases with MyD with abnormally elevated serum potassium; 2 of the 3 cases presented clinically with cardiac dysrhythmias. Hyperkalaemic conditions such as renal failure, cortisol deficiency, pseudohyperkalaemia, and hyperkalaemic periodic paralysis were excluded. Further endocrine evaluation revealed baseline hypoaldosteronism associated with elevated renin activity. Perturbation of the renin-angiotensin-aldosterone system resulted in appropriately enhanced renin activity but with a subnormal aldosterone response, which appeared to be due to adrenal hyporesponsiveness. The treatment of all cases with fludrocortisone was without effect. Whether the apparent mineralocorticoid abnormality in MyD is due to associated hormonal perturbations (i.e. excessive ACTH responsiveness. elevated cytokines, elevated atrial natriuretic hormone, etc.), adrenal atrophy, and/or a manifestation of the underlying kinase dysfunction is uncertain, but merits further evaluation in view of the clinical consequence of hyperkalaemia. PMID- 11874423 TI - Maternal transmission of diabetes. AB - Type 2 diabetes mellitus represents a heterogeneous group of conditions characterized by impaired glucose homeostasis. The disorder runs in families but the mechanism underlying this is unknown. Many, but not all, studies have suggested that mothers are excessively implicated in the transmission of the disorder. A number of possible genetic phenomena could explain this observation, including the exclusively maternal transmission of mitochondrial DNA (mtDNA). It is now apparent that mutations in mtDNA can indeed result in maternally inherited diabetes. Although several mutations have been implicated, the strongest evidence relates to a point substitution at nucleotide position 3243 (A to G) in the mitochondrial tRNA(leu(UUR)) gene. Mitochondrial diabetes is commonly associated with nerve deafness and often presents with progressive non-autoimmune beta-cell failure. Specific treatment with Coenzyme Q10 or L-carnitine may be beneficial. Several rodent models of mitochondrial diabetes have been developed, including one in which mtDNA is specifically depleted in the pancreatic islets. Apart from severe, pathogenic mtDNA mutations, common polymorphisms in mtDNA may contribute to variations of insulin secretory capacity in normal individuals. Mitochondrial diabetes accounts for less than 1% of all diabetes and other mechanisms must underlie the maternal transmission of Type 2 diabetes. Possibilities include the role of maternally controlled environments, imprinted genes and epigenetic phenomena. PMID- 11874424 TI - Risk of diabetes-related amputation in South Asians vs. Europeans in the UK. AB - AIMS: To compare the risk of and risk factors for diabetes-related amputation in South Asians and Europeans. METHODS: This was a population-based case control study based in the health districts of Bolton, Oldham and Central Manchester in the UK. Cases with diabetes-related amputation performed between 1992 and 1997 (n = 172) and controls with diabetes and no amputation (n = 376) were selected from the primary care-based North-west Diabetes Foot Study database. Risk factor data were also collected. RESULTS: Age at diagnosis adjusted odds ratio (OR) of amputation in South Asians compared with Europeans was 0.26, 95% confidence interval (CI) 0.11-0.65, P = 0.004. In the control population, South Asians were less likely than Europeans to have peripheral vascular disease (PVD) (9% vs. 24%, P = 0.02), neuropathy (30% vs. 54%, P = 0.003), and less likely to have ever been smokers (31% vs. 57%, P = 0.03). When these factors were added to the model, the OR was attenuated to 0.84, 95% CI 0.23-3.08, P = 0.8. CONCLUSIONS: South Asians with diabetes have about a quarter of the risk of amputation of Europeans. This is mostly explained by low rates of PVD and neuropathy in South Asians, in part associated with low rates of smoking. The reasons for the South Asian protection from both PVD and neuropathy deserve further exploration. PMID- 11874425 TI - Automated detection of diabetic retinopathy on digital fundus images. AB - AIMS: The aim was to develop an automated screening system to analyse digital colour retinal images for important features of non-proliferative diabetic retinopathy (NPDR). METHODS: High performance pre-processing of the colour images was performed. Previously described automated image analysis systems were used to detect major landmarks of the retinal image (optic disc, blood vessels and fovea). Recursive region growing segmentation algorithms combined with the use of a new technique, termed a 'Moat Operator', were used to automatically detect features of NPDR. These features included haemorrhages and microaneurysms (HMA), which were treated as one group, and hard exudates as another group. Sensitivity and specificity data were calculated by comparison with an experienced fundoscopist. RESULTS: The algorithm for exudate recognition was applied to 30 retinal images of which 21 contained exudates and nine were without pathology. The sensitivity and specificity for exudate detection were 88.5% and 99.7%, respectively, when compared with the ophthalmologist. HMA were present in 14 retinal images. The algorithm achieved a sensitivity of 77.5% and specificity of 88.7% for detection of HMA. CONCLUSIONS: Fully automated computer algorithms were able to detect hard exudates and HMA. This paper presents encouraging results in automatic identification of important features of NPDR. PMID- 11874426 TI - Diabetic nephropathy is associated with the 5'-end dinucleotide repeat polymorphism of the aldose reductase gene in Chinese subjects with Type 2 diabetes. AB - AIMS: We investigated whether the promoter dinucleotide repeat polymorphism of the aldose reductase gene (5'-ALR2), implicated in the development of nephropathy in Type 1 diabetes, was associated with diabetic nephropathy in Type 2 diabetes. METHODS: In 265 Southern Chinese with Type 2 diabetes the 5' -ALR2 polymorphism was identified in genomic DNA using polymerase chain reaction and automated fluorescent scanning. They were classified as normoalbuminuric (n = 128), microalbuminuric (n = 85) or albuminuric (n = 52) according to the mean albumin excretion rate of two 12-h overnight collections. RESULTS: The 5' -ALR2 allele and genotype distributions differed significantly among the three groups of patients (P < 0.003 and P < 0.01, respectively). Normoalbuminuric patients had the lowest Z - 2 allele frequency: 17.6% vs. 28.2% and 23.1% for microalbuminuric and albuminuric patients, respectively, and the highest Z + 2 allele frequency: 36.7% vs. 21.2% and 23.1% in microalbuminuric and albuminuric patients, respectively. They also had the lowest Z - 2/X genotype frequency (X = any allele other than Z + 2): 18.8% vs. 36.5% in microalbuminuric (P < 0.01) and 38.5% in albuminuric patients (P < 0.02), respectively, but the highest Z + 2/Y genotype frequency (Y = any allele other than Z - 2): 50.7% vs. 27.0% and 34.6% in microalbuminuric (P < 0.001) and albuminuric patients, respectively. In a multiple logistic regression model, the Z - 2/X genotype (odds ratio 3.10; P < 0.025) was an independent risk factor of diabetic nephropathy, microalbuminuria or albuminuria, together with age, mean arterial pressure and body mass index. CONCLUSIONS: The 5' -ALR2 dinucleotide repeat polymorphism is associated with the development of diabetic nephropathy in Southern Chinese with Type 2 diabetes. PMID- 11874427 TI - Role of sibutramine in the treatment of obese Type 2 diabetic patients receiving sulphonylurea therapy. AB - AIMS: To investigate whether the satiety-inducing agent sibutramine affected body weight and associated anthropometry in overweight and obese (body mass index (BMI) > 27) Type 2 diabetic patients on sulphonylurea therapy. METHODS: A randomized, placebo-controlled trial was undertaken in 134 patients with stable metabolic control on chronic sulphonylurea therapy. Patients were placed on moderate caloric restriction and received treatment with either sibutramine (15 mg/day) or placebo for 6 months. RESULTS: Fifty-three of 69 sibutramine-treated and 57/65 placebo-treated patients completed the study. Both groups showed progressive weight loss. At the end of the trial weight loss was two times greater in the sibutramine group (mean +/- SEM; -4.5 +/- 0.5 kg) than placebo ( 1.7 plus minus 0.5 kg, P < 0.001 vs. sibutramine). There was a trend for more patients to lose > 5% of initial body weight in the sibutramine group than placebo. BMI (P < 0.001) and waist circumference (P < 0.001) were also decreased to a greater extent by sibutramine. Mean reductions in HbA(1c) were commensurate with weight loss in both the sibutramine and placebo (- 0.78 +/- 0.17% and -0.73 +/- 0.23%; P = 0.84). Sibutramine was well tolerated with only two patients withdrawn due to potentially drug-related serious adverse events (palpitations). CONCLUSIONS: Sibutramine, in conjunction with moderate caloric restriction, enhances weight loss and reduces waist circumference in overweight and obese Type 2 diabetic patients receiving sulphonylurea therapy. This is associated with additional improvements in glycaemic control in a limited number of patients losing > or = 10% of their baseline body weight. PMID- 11874428 TI - Glycaemic control, disease duration and beta-cell function in patients with Type 2 diabetes in a Swedish community. Skaraborg Hypertension and Diabetes Project. AB - AIMS: To examine determinants for glycaemic control in primary care patients with Type 2 diabetes. METHODS: In a community-based surveillance of primary care patients with Type 2 diabetes, 190 men and 186 women were consecutively identified and examined for cardiovascular risk factors. Insulin resistance and beta-cell function were estimated using homeostasis model assessment (HOMA). Good glycaemic control was defined as HbA(1c) < 6.5%. RESULTS: Following adjustment for age and gender, HbA(1c) > or = 6.5% was associated with duration of diabetes (10.6 vs. 6.4 years, P < 0.001), lower levels of serum insulin (6.3 vs. 8.0 mU/l, P = 0.012), higher serum triglyceride levels (2.0 vs. 1.7 mmol/l, P = 0.002) and impairment of beta-cell function (HOMA index 19.5 vs. 45.8, P < 0.001). The association between HbA(1c) levels and duration remained with adjustment for age, gender, waist-hip ratio (WHR) and serum triglycerides (odds ratio (OR) for HbA(1c) > or = 6.5% by 5 years diabetes duration = 1.7; 95% confidence interval (CI) 1.4--2.1) but was lost following additional adjustment for beta-cell function (OR for HbA(1c) > or = 6.5% = 1.3; 95% CI 0.96-1.7). In a separate linear regression with beta-cell function as the dependent variable there was a significant association with HbA1c after adjustments for differences in age, gender, WHR, serum triglyceride levels and diabetes duration (P < 0.001). CONCLUSIONS: Increasing HbA1c by time was associated with declining beta-cell function. PMID- 11874429 TI - Impact of poverty on the prevalence of diabetes and its complications in urban southern India. AB - AIM: The impact of poverty on the profile of diabetes and its complications was studied. METHODS: A comparative study of low income group (LIG) (family income Rs. < 30,000/annum (approx. 432 pounds sterling) and high income group (HIG) (family income Rs. greater-than-or-equal 60,000/annum (approx. pounds sterling) subjects of > or = 40 years was done in Madras, India. By screening 1748 LIG subjects (M/W 844/904) 301 diabetic subjects were identified and 218 underwent tests for diabetic complications. Population data available in 635 (M/W 309/326) HIG subjects from the survey were used for comparison of glucose tolerance profile. Complications were studied in 221 diabetic HIG subjects. RESULTS: Age standardized prevalences of diabetes (12.6% vs. 25.5%; chi(2) = 56.9, P < 0.0001) and impaired glucose tolerance (IGT) (8.9% vs. 19.0%) were significantly lower (chi(2) = 57.7; P < 0.0001) in the LIG. Hypertension was more common in LIG (53.7% vs. 40.0% in HIG; chi(2) = 34.9; P < 0.0001). LIG subjects were more physically active; 73.8% did not go to school. Parameters significantly associated with diabetes were body mass index (BMI), age, higher income, waist- hip ratio and physical inactivity. Higher income, BMI and age were associated with IGT. Diabetic LIG subjects had a higher prevalence of cardiac disease, neuropathy and cataract and a lower prevalence of retinopathy than HIG subjects. The risk variables such as hyperglycaemia, dyslipidaemia, hypertension, smoking and alcohol consumption were more in the LIG group. CONCLUSIONS: The urban poor in the developing world has a lower prevalence of diabetes than the urban poor in developed societies. However, they have higher rates of complications of diabetes. PMID- 11874430 TI - Vascular effects of glibenclamide vs. glimepiride and metformin in Type 2 diabetic patients. AB - AIMS: Glibenclamide attenuates the protective responses to opening of vascular ATP-sensitive potassium (K(ATP)) channels during ischaemia. Therefore, glibenclamide treatment of Type 2 diabetes mellitus may have hazardous cardiovascular effects when used under conditions of ischaemia. Glimepiride and metformin seem to lack such characteristics. Based on these data, we hypothesized that, in contrast to glibenclamide, chronic treatment of Type 2 diabetic patients with glimepiride or metformin will not impair the vasodilator function of K(ATP) opening in vivo. METHODS: Two groups of 12 Type 2 diabetes mellitus patients participated in a double-blind randomized cross-over study consisting of two 8 week periods, in which treatment with orally administered glibenclamide (15 mg/day) was compared with either glimepiride or metformin (6 mg and 1500 mg/day, respectively). At the end of each treatment period, the increase in forearm blood flow (FBF, venous occlusion plethysmography) in response to intra-arterial administered diazoxide (K(ATP) opener), acetylcholine (endothelium-dependent vasodilator) and dipyridamole (adenosine-uptake blocker) and to forearm ischaemia was measured. RESULTS: There were no significant differences in vasodilator responses to diazoxide, acetylcholine, dipyridamole and forearm ischaemia after glibenclamide compared with glimepiride and metformin. CONCLUSIONS: Chronic treatment of Type 2 diabetes mellitus with glimepiride or metformin has similar effects on vascular K(ATP) channels compared with chronic glibenclamide treatment. PMID- 11874431 TI - Insulin, insulin propeptides and intima-media thickness in the carotid artery in 58-year-old clinically healthy men. The Atherosclerosis and Insulin Resistance study (AIR). AB - AIMS: To examine the relationship between specific (intact) insulin, insulin propeptides and subclinical atherosclerosis. METHODS: A cross-sectional study based on a stratified sampling of randomly selected, clinically healthy 58-year old men (n = 391). Ultrasound examinations of the carotid arteries were performed with measurement of intima-media thickness (IMT) in the common carotid artery and in the carotid artery bulb. Fasting, cross-reacting plasma insulin (RIA), specific (intact) insulin, proinsulin, 32,33 split proinsulin and C-peptide were measured. RESULTS: Plasma concentrations of cross-reacting plasma insulin, specific insulin, proinsulin, 32,33 split proinsulin and C-peptide were univariately associated with common carotid artery IMT. Established risk factors such as blood pressure, smoking, apoB, triglycerides, body mass index (BMI), and waist--hip ratio were also related to IMT. After adjustment for smoking, apoB, blood pressure and triglycerides, cross-reacting plasma insulin, proinsulin and C peptide but not specific insulin and split 32,33 proinsulin remained associated with carotid artery IMT. No associations remained after adjustment for BMI. CONCLUSIONS: Fasting plasma proinsulin, C-peptide, and insulin by cross-reacting RIA was associated with common carotid artery IMT independent of several conventional risk factors for atherosclerosis. The multicollinearity between the insulin peptides and propeptides makes it difficult to clarify the exact role of each peptide. PMID- 11874432 TI - Rheological determinants of red blood cell aggregation in diabetic patients in relation to their metabolic control. AB - AIMS: To determine whether increased red blood cell adhesiveness/aggregation in diabetic patients is related to the extent of their metabolic control. METHODS: We measured erythrocyte adhesiveness/aggregation in a group of 85 adult patients with diabetes mellitus by using citrated venous whole blood and a simple slide test. The erythrocyte adhesiveness/aggregation was determined by measuring the size of the spaces that are formed between the aggregated erythrocytes. We divided the patients into those with either low or high erythrocyte adhesiveness/aggregation values. RESULTS: The erythrocyte adhesiveness/aggregation values of the two groups differed significantly in terms of their fibrinogen concentration, erythrocyte sedimentation rate, high sensitive C-reactive protein (CRP), total cholesterol and triglyceride concentrations. There was no difference between the two groups regarding the concentrations of HbA(1c). Logistic regression was applied to construct a model to predict the belonging of a patient in the low or high erythrocyte adhesiveness/aggregation group. A linear regression was applied to construct a model to predict the erythrocyte adhesiveness/aggregation values. Both models turned out to include gender, age, fibrinogen, triglyceride, retinopathy, coronary artery disease and age and gender interaction. Neither HbA(1c) nor CRP entered the models. CONCLUSIONS: The degree of erythrocyte adhesiveness/aggregation and several variables of the acute-phase response in patients with diabetes mellitus are not directly related to the degree of metabolic control as evaluated by means of HbA(1c) concentration. Diabetic patients might benefit from rheological or anti inflammatory interventions regardless of their metabolic control. PMID- 11874433 TI - Blood Glucose Awareness Training in Dutch Type 1 diabetes patients. Short-term evaluation of individual and group training. AB - AIMS: The aims of the present study were: (i) to evaluate the effects of a Dutch translation and adaptation of Blood Glucose Awareness Training (BGAT-III) on blood glucose (bg) perception, glycaemic control, and decisions not to drive or to raise the bg during hypoglycaemia; (ii) to compare the effects of individual and group BGAT. METHODS: Fifty-nine patients with Type 1 diabetes participated in BGAT in either a group or an individual setting. Before and after BGAT, 39 (66%) of them completed 30-70 measurements on a hand-held computer (hhc). During every measurement, they estimated their bg, indicated whether they would drive or raise their bg on the basis of their estimation, and then measured their bg. RESULTS: Individual and group BGAT did not have significantly different effects (P = 0.35 0.98). Overall, BGAT did not significantly affect bg perception (P = 0.11-0.65). Before BGAT patients recognized a mean of 32% of their hypoglycaemic episodes, after BGAT a mean of 39% (P = 0.12). After BGAT, patients more often decided not to drive when their bg was low (P = 0.03). They tended to decide more often to raise their bg during hypoglycaemia (P = 0.09). CONCLUSIONS: The effects of BGAT were smaller than expected. Possible reasons for this negative outcome may be the adapted version of BGAT (shorter in duration), a lack of statistical power, or a difference between American and European samples in their reaction to BGAT. PMID- 11874434 TI - Trends in the incidence of childhood diabetes in south Asians and other children in Bradford, UK. AB - AIMS: To investigate incidence rates and time trends, over 21 years, of Type 1 diabetes in a migrant population of south Asian children in Bradford, UK. METHODS: Children (0-14 years) living in the city of Bradford and diagnosed with Type 1 diabetes were selected from a population-based region-wide register. Between 1978 and 1998, 289 new-onset cases were registered and classified as south Asian (Indian, Pakistani, Bangladeshi) or not, based on their full name using two different computer algorithms and visual inspection. RESULTS: Sixty-six children (22.8%) were designated as south Asian with 223 (77.2%) remaining. The overall age-sex standardized incidence for south Asian and non-south Asian children was 13.0 per 100,000 person years (95% confidence interval 9.9-16.2) and 12.9 (11.2-14.6), respectively. Rates were similar for south Asians at all ages, whereas for the mainly Caucasian children incidence differed significantly by age group (P < 0.001). An average annual increase in incidence of 4.3% (P = 0.001) was seen for all children compared with 6.5% in south Asians (P = 0.002) and 2.4% (P = 0.128) in non-south Asians. CONCLUSIONS: Children in south Asia have a low incidence of Type 1 diabetes but migrants to the UK have similar overall rates to the indigenous population. However, a more steeply rising incidence is seen in the south Asian population, and our data suggest that incidence in this group may eventually outstrip that of the non-south Asians. Genetic factors are unlikely to explain such a rapid change, implying an influence of environmental factors in disease aetiology. The similarity in rates by age group in the south Asian population is notable. PMID- 11874435 TI - Risk of developing diabetes is inversely related to lung function: a population based cohort study. AB - AIM: To investigate whether reduced lung function is a risk factor for developing diabetes. METHODS: Non-diabetic men (n = 382) from the population-based cohort 'Men Born in 1914' were examined with spirometry at age 55 years. The cohort was re-examined at 68 years. Diabetes and fasting plasma glucose at follow-up were studied in relation to vital capacity (VC) and forced expiratory volume (FEV1.0) at baseline. RESULTS: Fifteen men developed diabetes during the follow-up. The percentage with diabetes in the 1st, 2nd, 3rd and top quartile of vital capacity were 7%, 5%, 2%, and 1%, respectively (P for trend = 0.01). Fasting glucose (log transformed, mmol/l) at follow-up was 1.63 +/- 0.16, 1.62 +/- 0.18, 1.61 +/- 0.11 and 1.60 +/- 0.11, respectively (P for trend = 0.11). The longitudinal associations between VC and diabetes (P = 0.001) and log glucose (P = 0.036) were significant after adjustments for several potential confounders. FEV(1.0) at baseline showed similar associations with diabetes at follow-up. CONCLUSIONS: The risk of developing diabetes is inversely associated with pulmonary function among middle-aged men. PMID- 11874436 TI - Glucose-6-phosphate dehydrogenase: a candidate gene for diabetes. PMID- 11874437 TI - Eczema and Type 1 diabetes. PMID- 11874438 TI - Approaches to preventing mealtime hyperglycaemic excursions. PMID- 11874439 TI - Lipoic acid increases glucose uptake by skeletal muscles of obese-diabetic ob/ob mice. AB - AIM: Alpha-lipoic acid has been reported to increase glucose disposal in diabetic states. This study has examined the effect of alpha-lipoic acid on glucose uptake by cultured L6 muscle cells and different types of skeletal muscles in normal lean (+/+) and severely insulin-resistant, obese-diabetic (ob/ob) mice. METHODS: Glucose uptake was measured in L6 muscle cells using the non-metabolized glucose analogue 2-deoxy-d-glucose (2DG), and in isolated muscles by glucose disappearance from the incubation medium. RESULTS: In L6 muscle cells, short-term incubations (2-12 h) with 10(-3) m alpha-lipoic acid increased glucose uptake by 40-80%, approximately the same extent as 10(-6) m insulin. Combination of the two agents produced a slightly greater increase (120% at 12 h) than either alone. Red quadriceps (mainly type 1 fibres), diaphragm (similar proportions of type 1 and 2 fibres) and abdominal muscle (mainly type 2 fibres) from normal mice incubated with 10(-3) m alpha-lipoic acid showed increased glucose uptake to a similar extent as 10(-6) m insulin in each of the three muscles. Muscles from ob/ob mice, which showed little response to insulin, showed a substantial increase (approximately 300%, p < 0.05-0.01) in glucose uptake when 10(-3) m alpha-lipoic acid was added in the presence of insulin. The alpha-lipoic acid also increased glucose uptake in red quadriceps (approximately 300%, p < 0.01) from ob/ob mice without added insulin. CONCLUSION: The results suggest that alpha-lipoic acid can increase glucose uptake by a range of normal muscle types and improve the response to insulin by insulin-resistant skeletal muscles of ob/ob mice. PMID- 11874440 TI - Metformin improves lipid metabolism and attenuates lipid peroxidation in high fructose-fed rats. AB - AIM: Insulin resistance, hyperinsulinaemia and disturbances in glucose metabolism can be produced in normal rats by feeding them a fructose-enriched diet. Metformin, an antidiabetic drug, enhances insulin sensitivity in type 2 diabetic patients. Previous studies have shown that metformin improves insulin sensitivity in fructose-fed rats. The aim of this study was to determine the effect of metformin treatment on overall lipid metabolism and lipid peroxidation in rats that were fed a fructose-enriched diet, which leads to insulin resistance. The relationship between hyperinsulinaemia and hyperglycaemia with lipid peroxide levels was also investigated. MATERIALS AND METHODS: The animals were divided into two batches. One batch received a standard diet and the other was fed a fructose-enriched diet (72 g/100 g feed). After 2 weeks, each batch of rats was subdivided into two groups. One group received metformin (50 mg/kg per day in water) and the other received the standard diet and served as control. RESULTS: High fructose feeding resulted in hyperinsulinaemia, hyperglycaemia and alterations in lipids and lipid metabolism, and plasma and tissue lipid peroxides were significantly elevated. Administration of metformin (50 mg/kg/day) was associated with significant normalization of plasma insulin level and lipid alterations. These rats also showed significantly higher lipoprotein lipase (LPL) and lecithin cholesterol acyl transferase (LCAT) activities in plasma than untreated, fructose-fed rats. Lipid peroxides content was also decreased in plasma and tissues. Significant positive correlations were observed between the levels of plasma insulin and plasma glucose with plasma lipid peroxides. CONCLUSIONS: We conclude that enhanced lipid peroxidation occurs in addition to disturbances in lipid metabolism in fructose-fed rats. Plasma insulin level probably contributes to this increased peroxidation. Improved insulin action in metformin-treated rats could be responsible for the amelioration of these abnormalities induced by fructose feeding. PMID- 11874441 TI - Effects of a water-soluble extract of maitake mushroom on circulating glucose/insulin concentrations in KK mice. AB - AIM: We examined benefits of a water-soluble extract of maitake mushroom designated as Fraction X (FXM) on the glucose/insulin metabolism of insulin resistant KK mice, and compared the results of FXM with those of a sulphonylurea, Glipizide. DESIGN: In several acute studies, insulin-resistant KK mice were gavaged with a single dose of varying concentrations of FXM, or a single dose of one concentration of the oral hypoglycaemic drug, Glipizide. In the one chronic study, KK mice were gavaged with FXM, Glipizide, or an equal volume of isotonic saline (baseline control) twice daily. Retro-orbital blood was drawn on the morning of the 4th and 7th days before the early gavage. Blood glucose was measured by routine laboratory procedures, and serum insulin was estimated by a radioimmunoassay (RIA) assay developed specifically for rodents. RESULTS: At a dose of FXM (140 mg/mouse), a statistically significant lowering of circulating glucose concentrations was again seen at 8-12 h and 16-18 h after oral gavage. The lowering approximated 25% of the original concentration. Oral gavage of Glipizide resulted in statistically significantly lower values of circulating glucose (25-37% lower compared with baseline) at 8-24 h post dosing. In the chronic study, the circulating concentrations of glucose and insulin of mice taking 140 mg FXM per day were decreased significantly at days 4 and 7. CONCLUSIONS: FXM, a natural extract obtained from maitake mushroom, favourably influences glucose/insulin metabolism in insulin-resistant KK mice. The lowering of both circulating glucose and insulin concentrations suggests that FXM works primarily by enhancing peripheral insulin sensitivity. PMID- 11874442 TI - Evaluation of the safety and efficacy of sibutramine, orlistat and metformin in the treatment of obesity. AB - AIM: Some of our obese patients who were receiving 10 mg/day sibutramine reported feeling hunger at night. To address this, we designed a randomized, prospective clinical trial to study the efficacy and safety of 10 mg sibutramine twice daily (bid), and compare this treatment with 120 mg orlistat three times daily (tid) and 850 mg metformin (bid). METHODS: A total of 150 female patients with body mass index (b.m.i.) > 30 kg/m(2) were included. The subjects were all out patients at the Baskent University Endocrinology and Metabolism Clinic. Each individual was assigned randomly to receive 10 mg sibutramine bid (group 1; n = 50; mean age 42.27 +/- 1.40 years), 120 mg orlistat tid (group 2; n = 50; mean age 42.13 +/- 1.32 years) or 850 mg metformin bid (group 3; n = 50; mean age 43.58 +/- 1.40 years). All patients took the medications for 6 months. Two patients from the sibutramine group and two from the orlistat group were withdrawn from the study because of side-effects. RESULTS: After 6 months of treatment, the sibutramine, orlistat, and metformin groups all showed significantly reduced b.m.i. (13.57%, 9.06% and 9.90% respectively); waist circumference (10.43%, 6.64%, and 8.10% respectively); fasting and postprandial blood glucose levels; insulin resistance as assessed by the homeostasis model for assessment of insulin resistance (HOMA) (38.63%, 32.73% and 39.28%, respectively); levels of total cholesterol, low-density lipoprotein (LDLC) cholesterol, very low-density lipoprotein (VLDLC) cholesterol, triglyceride, lipoprotein (a), and apolipoprotein B; uric acid level; pulse rate; and systolic and diastolic blood pressure. None of the groups showed any significant changes in levels of high-density lipoprotein (HDLC) cholesterol, or apolipoprotein A1. There was a significantly greater fall in b.m.i. in the sibutramine group than in either of the other groups (p < 0.0001). CONCLUSIONS: The results of this study confirm that sibutramine, orlistat and metformin are all effective and safe medications that reduce cardiovascular risk and can decrease the risk of type 2 diabetes mellitus in obese females. Overall, treatment with 10 mg sibutramine bid is more effective than orlistat or metformin therapy in terms of weight reduction. PMID- 11874443 TI - Clinical evidence of thiazolidinedione-induced improvement of pancreatic beta cell function in patients with type 2 diabetes mellitus. AB - BACKGROUND: There is growing evidence in animal and in vitro studies showing that thiazolidinediones (TZDs) improve pancreatic beta cell (beta-cell) function. The aim of this study was to evaluate the effect of thiazolidinediones on the beta cell function of patients with Type 2 diabetes mellitus (T2DM) in clinical practice. PATIENTS AND METHODS: This is an observational, nested case-control study. We identified 28 patients (TZD group), with T2DM, who had had a meal stimulated C-peptide level documented before the addition of troglitazone to a failing double-therapy regimen with metformin (MET) and sulphonylurea (SU). As a control group (CTRL), we identified 26 patients, with T2DM, who also had had a meal-stimulated C-peptide documented before adding MET to a failing SU monotherapy regimen. We then proceeded to prospectively remeasure their meal stimulated C-peptide levels and compared the changes over time between the two groups. RESULTS: Both groups were well matched for age, body mass index (BMI), and HgbA1c before and after treatment. The C-peptide in the TZD group increased significantly during therapy (from 3.2 +/- 0.5 to 4.2 +/- 0.5, p = 0.01), whereas it remained unchanged in the CTRL group (from 4.8 +/- 0.6 to 5.0 +/- 0.5, p = 0.74). The C-peptide/glucose ratio also increased significantly in the TZD group (from 1.9 +/- 0.3 to 3.1 +/- 0.3, p = 0.0003) whereas it remained unchanged in the CTRL group (from 3.4 +/- 0.7 to 3.4 +/- 0.3, p = 0.97). Furthermore, the C peptide/glucose ratio of the TZD group, which was significantly lower at baseline compared with the CTRL group (1.9 +/- 0.3 vs. 3.4 +/- 0.7, p = 0.04), caught up to its level during treatment (3.1 +/- 0.3 vs. 3.4 +/- 0.3, p = 0.48). CONCLUSION: Thiazolidinediones seem to induce recovery of pancreatic beta cell function, independently of the correction of glucose toxicity. PMID- 11874444 TI - 2-[(18)F]fluoro-2-deoxy-D-glucose combined with microdialysis can be used for the comparison of tissue glucose metabolism in obese and lean rats. AB - AIMS: Direct assessment of tissue metabolism in vivo is important to understand the pathogenesis of obesity. Labelled glucose analogues are potential candidates to be used for this purpose. The aim of this study was to compare the kinetics and metabolism of 2-[(18)F]fluoro-2-deoxy-D-glucose (FDG) in obese (fa/fa) and lean (Fa/?) Zucker rat tissues with microdialysis, and the measurement of uptake and phosphorylation with or without insulin bolus injection. METHODS: Obese (n = 10) and lean (n = 11) anaesthetized rats underwent a microdialysis study after FDG-injection either with or without insulin stimulation. Microdialysis probes were inserted in the jugular vein, quadriceps muscle and liver. After 110 min, tissue [(18)F]-uptake and intracellular phosphorylation of FDG were studied in blood, liver, skeletal muscle, subcutaneous adipose tissue, intra-abdominal adipose tissue and hypothalamus. RESULTS: When measured with microdialysis, insulin-enhanced FDG disappeared from the blood pool and interstitial space of skeletal muscle and liver more effectively in lean rather than in obese animals. Insulin-stimulated skeletal muscle and adipose tissue[(18)F]-uptake was impaired in obese Zucker rats compared with lean animals. Hypothalamic FDG uptake was six to sevenfold higher than in other measured tissues, but was attenuated in obese rats. In liver and in mesenteric fat, insulin-enhanced FDG phosphorylation in lean rats compared with obese animals. CONCLUSIONS: Positron-emitting glucose analogue FDG, combined with microdialysis and tissue analysis, is a feasible method in studying glucose metabolism at the cellular level in animal studies. PMID- 11874445 TI - Elimination of *O(2)(-)/H(2)O(2) by alpha-lipoic acid mediates the recovery of basal EDRF/NO availability and the reversal of superoxide dismutase-induced relaxation in diabetic rat aorta. AB - AIM: The aims of this study were to ascertain the mechanism(s) of relaxant action of exogenous superoxide dismutase (SOD) in aortic rings obtained from 12-week, streptozotocin(STZ)-diabetic and age-matched control rats, and to examine the effects of alpha-lipoic acid (ALA) treatment (for 6 weeks, after 6 weeks of untreated diabetes) on SOD-induced relaxations. MATERIALS AND METHODS: Thoracic aorta rings were suspended to isolated tissue chamber, and the changes in isometric tension were recorded. RESULTS: SOD produced a greater relaxation in untreated-diabetic rings compared with control rings. ALA treatment partially reversed SOD-induced relaxation in diabetic aorta. Pretreatment of rings with N(G)-nitro-L-arginine methyl ester (L-NAME, 100 microm) inhibited SOD-induced relaxation. This effect of L-NAME was markedly observed in control and ALA treated-diabetic rings compared with untreated-diabetic rings. SOD-induced relaxation was also inhibited by catalase (60 U/ml) in untreated-diabetic rings but not in ALA-treated-diabetic and control rings. Pretreatment with the cyclooxygenase inhibitor, indomethacin, or the catalase inhibitor, aminotriazole, had no effect on SOD-induced relaxation in any ring. CONCLUSION: Findings suggested that: (i) in normal physiological conditions, the relaxant effect of SOD is related to the inhibition of superoxide anion radicals (*O(2)(-))-induced endothelium-derived relaxing factor/nitric oxide (EDRF/NO) destruction in the rat aorta; (ii) in diabetic state, excess *O(2)(-) increasingly inhibits basal EDRF/NO, and the dismutation of excess *O(2)(-) to H(2)O(2) is enhanced by exogenous SOD. H(2)O(2) a vasorelaxant molecule, which probably accounts for the increased responsiveness of diabetic rings to exogenous SOD; and (iii) the reversal effect of in vivo ALA treatment on SOD-induced relaxation in diabetic aorta is probably linked with the elimination of *O(2)(-)/H(2)O(2), which mediates the recovery of basal EDRF/NO availability. PMID- 11874446 TI - Effects of aminoguanidine and tolrestat on the development of ocular and renal structural changes in experimental diabetic rats. AB - Studies that researched the role of aminoguanidine and tolestat in the prevention of diabetic retinopathy and nephropathy resulted in conflicting data. We investigated the effects of these agents in the prevention of ocular and renal changes in streptozotocin (STZ)-induced diabetic rats. Diabetes was induced by intravenous injection of STZ in 30 rats. Ten rats that were not given STZ served as non-diabetic control (Group 1). Ten STZ-diabetic rats that were not given any treatment served as diabetic control (Group 2). Groups 3 and 4 were composed of STZ-induced diabetic rats (10 each) that were given tolrestat and aminoguanidine respectively. Eyes and kidneys were examined at the 24th week under electronmicroscopy. Cataract was observed in all six of the surviving rats in Groups 2 and 4, and in one of 6 surviving rats in group 3. Cataract development was lower in Group 3 than Groups 2 and 4. All retinal samples obtained from group 2 demonstrated a number of structural abnormalities, whereas there were no significant ultrastructural changes in groups 3 and 4. Groups 2 and 3 demonstrated mesangial proliferation and expansion, diffuse glomerular basement membrane (GBM) thickening, and focal GBM thickening in the bulb form. Group 4 demonstrated a normally appearing mesangial space, minimal diffuse but no focal GBM thickening. The urinary albumin excretion (UAE) was lower in Group 4 than the other groups. In conclusion, our results suggest that aminoguanidine may be an important agent for the prevention of renal changes, whereas tolrestat may be effective for the prevention of ocular changes in diabetes mellitus. PMID- 11874447 TI - Modelling of simple and complex calcium oscillations. From single-cell responses to intercellular signalling. AB - This review provides a comparative overview of recent developments in the modelling of cellular calcium oscillations. A large variety of mathematical models have been developed for this wide-spread phenomenon in intra- and intercellular signalling. From these, a general model is extracted that involves six types of concentration variables: inositol 1,4,5-trisphosphate (IP3), cytoplasmic, endoplasmic reticulum and mitochondrial calcium, the occupied binding sites of calcium buffers, and the fraction of active IP3 receptor calcium release channels. Using this framework, the models of calcium oscillations can be classified into 'minimal' models containing two variables and 'extended' models of three and more variables. Three types of minimal models are identified that are all based on calcium-induced calcium release (CICR), but differ with respect to the mechanisms limiting CICR. Extended models include IP3--calcium cross coupling, calcium sequestration by mitochondria, the detailed gating kinetics of the IP3 receptor, and the dynamics of G-protein activation. In addition to generating regular oscillations, such models can describe bursting and chaotic calcium dynamics. The earlier hypothesis that information in calcium oscillations is encoded mainly by their frequency is nowadays modified in that some effect is attributed to amplitude encoding or temporal encoding. This point is discussed with reference to the analysis of the local and global bifurcations by which calcium oscillations can arise. Moreover, the question of how calcium binding proteins can sense and transform oscillatory signals is addressed. Recently, potential mechanisms leading to the coordination of oscillations in coupled cells have been investigated by mathematical modelling. For this, the general modelling framework is extended to include cytoplasmic and gap-junctional diffusion of IP3 and calcium, and specific models are compared. Various suggestions concerning the physiological significance of oscillatory behaviour in intra- and intercellular signalling are discussed. The article is concluded with a discussion of obstacles and prospects. PMID- 11874449 TI - Prediction of protein--protein interaction sites in heterocomplexes with neural networks. AB - In this paper we address the problem of extracting features relevant for predicting protein--protein interaction sites from the three-dimensional structures of protein complexes. Our approach is based on information about evolutionary conservation and surface disposition. We implement a neural network based system, which uses a cross validation procedure and allows the correct detection of 73% of the residues involved in protein interactions in a selected database comprising 226 heterodimers. Our analysis confirms that the chemico physical properties of interacting surfaces are difficult to distinguish from those of the whole protein surface. However neural networks trained with a reduced representation of the interacting patch and sequence profile are sufficient to generalize over the different features of the contact patches and to predict whether a residue in the protein surface is or is not in contact. By using a blind test, we report the prediction of the surface interacting sites of three structural components of the Dnak molecular chaperone system, and find close agreement with previously published experimental results. We propose that the predictor can significantly complement results from structural and functional proteomics. PMID- 11874450 TI - Proteolysis of bovine beta-lactoglobulin during thermal treatment in subdenaturing conditions highlights some structural features of the temperature modified protein and yields fragments with low immunoreactivity. AB - Bovine beta-lactoglobulin was hydrolyzed with trypsin or chymotrypsin in the course of heat treatment at 55, 60 and 65 degrees C at neutral pH. At these temperatures beta-lactoglobulin undergoes significant but reversible structural changes. In the conditions used in the present study, beta-lactoglobulin was virtually insensitive to proteolysis by either enzyme at room temperature, but underwent extensive proteolysis when either protease was present during the heat treatment. High-temperature proteolysis occurs in a progressive manner. Mass spectrometry analysis of some large-sized breakdown intermediates formed in the early steps of hydrolysis indicated that both enzymes effectively hydrolyzed some regions of beta-lactoglobulin that were transiently exposed during the physical treatments and that were not accessible in the native protein. The immunochemical properties of the products of beta-lactoglobulin hydrolysis were assessed by using various beta-lactoglobulin-specific antibodies, and most epitopic sites were no longer present after attack of the partially unfolded protein by the two proteases. PMID- 11874451 TI - Uncoupling of protein-3 induces an uncontrolled uncoupling of mitochondria after expression in muscle derived L6 cells. AB - The uncoupling proteins (UCPs) are thought to uncouple oxidative phosphorylation in the mitochondria and thus generate heat. One of the UCP isoforms, UCP3, is abundantly expressed in skeletal muscle, the major thermogenic tissue in humans. UCP3 has been overexpressed at high levels in yeast systems, where it leads to the uncoupling of cell respiration, suggesting that UCP3 may indeed be capable of dissipating the mitochondrial proton gradient. This effect, however, was recently shown to be a consequence of the high level of expression and incorrect folding of the protein and not to its intrinsic uncoupling activity. In the present study, we investigated the properties of UCP3 overexpressed in a relevant mammalian host system such as the rat myoblast L6 cell line. UCP3 was expressed in relatively low levels (< 1 microg x mg(-1) membrane protein) with the help of an adenovirus vector. Immunofluorescence microscopy of transduced L6 cells showed that UCP3 was expressed in more than 90% of the cells and that its staining pattern was characteristic for mitochondrial localization. The oxygen consumption of L6 cells under nonphosphorylating conditions increased concomitantly with the levels of UCP3 expression. However, uncoupling was associated with an inhibition of the maximal respiratory capacity of mitochondria and was not affected by purine nucleotides and free fatty acids. Moreover, recombinant UCP3 was resistant to Triton X-100 extraction under conditions that fully solubilize membrane bound proteins. Thus, UCP3 can be uniformly overexpressed in the mitochondria of a relevant muscle-derived cell line resulting in the expected increase of mitochondrial uncoupling. However, our data suggest that the protein is present in an incompetent conformation. PMID- 11874453 TI - Study of substrate specificity of human aromatase by site directed mutagenesis. AB - Human aromatase is responsible for estrogen biosynthesis and is implicated, in particular, in reproduction and estrogen-dependent tumor proliferation. The molecular structure model is largely derived from the X-ray structure of bacterial cytochromes sharing only 15-20% identities with hP-450arom. In the present study, site directed mutagenesis experiments were performed to examine the role of K119, C124, I125, K130, E302, F320, D309, H475, D476, S470, I471 and I474 of aromatase in catalysis and for substrate binding. The catalytic properties of mutants, transfected in 293 cells, were evaluated using androstenedione, testosterone or nor-testosterone as substrates. In addition, inhibition profiles for these mutants with indane or indolizinone derivatives were obtained. Our results, together with computer modeling, show that catalytic properties of mutants vary in accordance with the substrate used, suggesting possible differences in substrates positioning within the active site. In this respect, importance of residues H475, D476 and K130 was discussed. These results allow us to hypothesize that E302 could be involved in the aromatization mechanism with nor-androgens, whereas D309 remains involved in androgen aromatization. This study highlights the flexibility of the substrate-enzyme complex conformation, and thus sheds new light on residues that may be responsible for substrate specificity between species or aromatase isoforms. PMID- 11874452 TI - Induction of (2'-5')oligoadenylate synthetase in the marine sponges Suberites domuncula and Geodia cydonium by the bacterial endotoxin lipopolysaccharide. AB - Recent studies have shown that the Porifera, with the examples of the demosponges Suberites domuncula and Geodia cydonium, comprise a series of pathways found also in the immune system of Deuterostomia, such as vertebrates, but are absent in Protostomia, with insects or nematodes as examples. One pathway is the (2' 5')oligoadenylate synthetase [(2-5)A synthetase] system. In the present study we show that crude extracts from tissue of S. domuncula collected from the sea display a considerable amount of (2-5)A synthetase activity; 16% of the ATP substrate is converted to the (2-5)A product, while tissue from specimens which were kept for 6 months in an aquarium shows only 1% of conversion. As aquarium animals show a lower bacterial load, those specimens were treated for the experiments with the bacterial endotoxin lipopolysaccharide (LPS); they responded to LPS with a stimulation of the (2-5)A synthetase activity. To monitor if this effect can be obtained also on the in vitro level, primmorphs which comprise proliferating and differentiating cells, were incubated with LPS. Extracts obtained from LPS-treated primmorphs also convert ATP to the (2-5)A products mediated by the synthetase. In parallel to this effect on protein level, LPS causes after an incubation period of 12 h also an increase in the steady-state level of the transcripts encoding the putative (2-5)A synthetase. It is postulated that in sponges the (2-5)A synthetase is involved in antimicrobial defense of the animals. PMID- 11874454 TI - Structure of the O-polysaccharide and classification of Proteus mirabilis strain G1 in Proteus serogroup O3. AB - The O-chain polysaccharide of the lipopolysaccharide (LPS) of a previously nonclassified strain of Proteus mirabilis termed G1 was studied by sugar analysis and 1H and 13C NMR spectroscopy, including 2D COSY, TOCSY, rotating-frame NOE (ROESY), H-detected 1H,13C HMQC, and heteronuclear multiple-bond correlation (HMBC) experiments. The following structure of the polysaccharide was established: [carbohydrate structure: see text] where D-GalA6(L-Lys) stands for N(alpha)-(D-galacturonoyl)-L-lysine. The structure of the O-polysaccharide of P. mirabilis G1 is similar, but not identical, to that of P. mirabilis S1959 and OXK belonging to serogroup O3. Immunochemical studies with P. mirabilis G1 and S1959 anti-(O-polysaccharide) sera revealed close LPS-based serological relatedness of P. mirabilis G1 and S1959, and therefore it was suggested to classify P. mirabilis G1 in serogroup O3 as a subgroup. P. mirabilis G1 and S1959 anti-(O polysaccharide) sera also cross-reacted with LPS of P. mirabilis strains from two other serogroups containing D-GalA6(L-Lys) in the O-polysaccharide or in the core region. PMID- 11874455 TI - Artocarpus hirsuta lectin. Differential modes of chemical and thermal denaturation. AB - Unfolding, inactivation and dissociation of the lectin from Artocarpus hirsuta seeds were studied by chemical (guanidine hydrochloride, GdnHCl) and thermal denaturation. Conformational transitions were monitored by intrinsic fluorescence and circular dichroism. The gradual red shift in the emission maxima of the native protein from 335 to 356 nm, change in the ellipticity at 218 nm and simultaneous decrease in the sugar binding activity were observed with increasing concentration of GdnHCl in the pH range between 4.0 and 9.0. The unfolding and inactivation by GdnHCl were partially reversible. Gel filtration of the lectin in presence of 1-6 m GdnHCl showed that the protein dissociates reversibly into partially unfolded dimer and then irreversibly into unfolded inactive monomer. Thermal denaturation was irreversible. The lectin loses activity rapidly above 45 degrees C. The exposure of hydrophobic patches, distorted secondary structure and formation of insoluble aggregates of the thermally inactivated protein probably leads to the irreversible denaturation. PMID- 11874456 TI - CK2(beta)tes gene encodes a testis-specific isoform of the regulatory subunit of casein kinase 2 in Drosophila melanogaster. AB - An earlier described CK2(beta)tes gene of Drosophila melanogaster is shown to encode a male germline specific isoform of regulatory beta subunit of casein kinase 2. Western-analysis using anti-CK2(beta)tes Ig revealed CK2(beta)tes protein in Drosophila testes extract. Expression of a CK2(beta)tes-beta galactosidase fusion protein driven by the CK2(beta)tes promoter was found in transgenic flies at postmitotic stages of spermatogenesis. Examination of biochemical characteristics of a recombinant CK2(beta)tes protein expressed in Escherichia coli revealed properties similar to those of CK2beta: (a) CK2(beta)tes protein stimulates CK2alpha catalytic activity toward synthetic peptide; (b) it inhibits phosphorylation of calmodulin and mediates stimulation of CK2alpha by polylysine; (c) it is able to form (CK2(beta)tes)2 dimers, as well as (CK2alpha)2(CK2(beta)tes)2 tetramers. Using the yeast two-hybrid system and coimmunoprecipitation analysis of protein extract from Drosophila testes, we demonstrated an association between CK2(beta)tes and CK2alpha. Northern-analysis has shown that another regulatory (beta') subunit found recently in D. melanogaster genome is also testis-specific. Thus, we describe the first example of two tissue-specific regulatory subunits of CK2 which might serve to provide CK2 substrate recognition during spermatogenesis. PMID- 11874457 TI - Interaction of decorin with CNBr peptides from collagens I and II. Evidence for multiple binding sites and essential lysyl residues in collagen. AB - Decorin is a small leucine-rich chondroitin/dermatan sulfate proteoglycan reported to interact with fibrillar collagens through its protein core and to localize at d and e bands of the collagen fibril banding pattern. Using a solid phase assay, we have determined the interaction of peptides derived by CNBr cleavage of type I and type II collagen with decorin extracted from bovine tendon and its protein core and with a recombinant decorin preparation. At least five peptides have been found to interact with all three decorin samples. The interaction of peptides with tendon decorin has a dissociation constant in the nanomolar range. The triple helical conformation of the peptide trimeric species is a necessary requisite for the binding. All positive peptides have a region within the d and e bands of collagen fibrils. Two chemical derivatives of collagens and of positive peptides were prepared by N-acetylation and N methylation of the primary amino group of Lys/Hyl side chains. Chemical modifications performed in mild conditions do not significantly alter the thermal stability of peptide trimeric species whereas they affect the interaction with decorin: N-acetylation eliminates both the positive charge and the binding to decorin, whereas N-methylation preserves the cationic character and modulates the binding. We conclude that decorin makes contacts with multiple sites in type I collagen and probably also in type II collagen and that some collagen Lys/Hyl residues are essential for the binding. PMID- 11874458 TI - The insert within the catalytic domain of tripeptidyl-peptidase II is important for the formation of the active complex. AB - Tripeptidyl-peptidase II (TPP II) is a large (Mr>10(6)) tripeptide-releasing enzyme with an active site of the subtilisin-type. Compared with other subtilases, TPP II has a 200 amino-acid insertion between the catalytic Asp44 and His264 residues, and is active as an oligomeric complex. This study demonstrates that the insert is important for the formation of the active high-molecular mass complex. A recombinant human TPP II and a murine TPP II were found to display different complex-forming characteristics when over-expressed in human 293-cells; the human enzyme was mainly in a nonassociated, inactive state whereas the murine enzyme formed active oligomers. This was surprising because native human TPP II is purified from erythrocytes as an active oligomeric complex, and the amino-acid sequences of the human and murine enzymes were 96% identical. Using a combination of chimeras and a single point mutant, the amino acid responsible for this difference was identified as Arg252 in the recombinant human sequence, which corresponds to a glycine in the murine sequence. As Gly252 is conserved in all sequenced variants of TPP II, the recombinant enzyme with Arg252 is atypical. Nevertheless, as Arg252 evidently interferes with complex formation, and this residue is close to the catalytic His264, it may also explain why oligomerization influences enzyme activity. The exact mechanism for how the G252R substitution interferes with complex formation remains to be determined, but will be of importance for the understanding of the unique properties of TPP II. PMID- 11874459 TI - NMR-based determination of the binding epitope and conformational analysis of MUC 1 glycopeptides and peptides bound to the breast cancer-selective monoclonal antibody SM3. AB - Mucin glycoproteins on breast cancer cells carry shortened carbohydrate chains. These partially deglycosylated mucin 1 (MUC-1) structures are recognized by the monoclonal antibody SM3, which is being tested for its diagnostic utility. We used NMR spectroscopy to analyze the binding mode and the binding epitope of peptide and glycopeptide antigens to the SM3 antibody. The pentapeptide PDTRP and the glycopentapeptide PDT(O-alpha-D-GalNAc)RP are known ligands of the monoclonal antibody. The 3D structures of the ligands in the bound conformation were determined by analyzing trNOESY build-up rates. The peptide was found to adopt an extended conformation that fits into the binding pocket of the antibody. The binding epitopes of the ligands were determined by saturation transfer difference (STD) NMR spectroscopy. The peptide's epitope is predominantly located in the N terminal PDT segment whereas the C-terminal RP segment has fewer interactions with the protein. In contrast, the glycopeptide is interacting with SM3 utilizing all its amino acids. Pro1 shows the strongest binding effect that slightly decays towards Pro5. The GalNAc residue interacts mainly via the N-acetyl residue while the other protons show less interactions similar to that of Pro5. The glycopeptide in the bound state also has an extended conformation of the peptide with the carbohydrate oriented towards the N-terminus. Docking studies showed that peptide and glycopeptide fit the binding pocket of the mAb SM3 very well. PMID- 11874460 TI - Overexpression of a recombinant wild-type and His-tagged Bacillus subtilis glycine oxidase in Escherichia coli. AB - We have cloned the gene coding for the Bacillus subtilis glycine oxidase (GO), a new flavoprotein that oxidizes glycine and sarcosine to the corresponding alpha keto acid, ammonia and hydrogen peroxide. By inserting the DNA encoding for GO into the multiple cloning site of the expression vector pT7.7 we produced a recombinant plasmid (pT7-GO). The pT7-GO encodes a fully active fusion protein with six additional residues at the N-terminus of GO (MARIRA). In BL21(DE3)pLysS Escherichia coli cells, and under optimal isopropyl thio-beta-D-galactoside induction conditions, soluble and active chimeric GO was expressed up to 1.14 U g(-1) of cell (and a fermentation yield of 3.82 U x L(-1) of fermentation broth). An N-terminal His-tagged protein (HisGO) was also successfully expressed in E. coli as a soluble protein and a fully active holoenzyme. HisGO represents approximately 3.9% of the total soluble protein content of the cell. The His tagged GO was purified in a single step by nickel-chelate chromatography to a specific activity of 1.06 U x mg(-1) protein at 25 degrees C and with a yield of 98%. The characterization of the purified enzyme showed that GO is a homotetramer of approximately 180 kDa with the spectral properties typical of flavoproteins. GO exhibits good thermal stability, with a Tm of 46 degrees C after 30 min incubation; its stability is maximal in the 7.0-8.5 pH range. A comparison of amino-acid sequence and substrate specificity indicates that GO has similarities to other flavoenzymes acting on primary amines and on D-amino acids. PMID- 11874461 TI - Regulation of alpha1,3galactosyltransferase expression in pig endothelial cells. Implications for xenotransplantation. AB - The disaccharide galactose(alpha)1,3 galactose (the alphaGal epitope) is the major xenoantigen responsible for the hyperacute vascular rejection occurring in pig-to-primates organ transplantation. The synthesis of the alphaGal epitope is catalyzed by the enzyme alpha1,3-galactosyltransferase (alpha1,3GalT). To be able to control porcine alpha1,3GalT gene expression specifically, we have analyzed the upstream portion of the alpha1,3GalT gene, and identified the regulatory sequences. Porcine alpha1,3GalT transcripts were detected by 5' RACE analysis, and the corresponding genomic sequences were isolated from a phage library. The porcine alpha1,3GalT gene consists of at least 10 different exons, four of which contain 5' untranslated sequence. Four distinct promoters, termed A-D, drive alpha1,3GalT gene transcription in porcine cells. A combination of alternative promoter usage and alternative splicing produces a series of transcripts that differ in their 5' portion, but encode the same protein. Promoters A-C have been isolated, and functionally characterized using luciferase reporter gene assays in transfected porcine endothelial cells (PEC-A). Promoter preference in porcine endothelial cells was estimated on the basis of relative transcript levels as determined by real-time quantitative PCR. More than 90% of the alpha1,3GalT transcripts in PEC-A cells originate from promoter B, which has characteristics of a housekeeping gene promoter. While promoter preference remains unchanged, alpha1,3GalT mRNA levels increase by 50% in 12 h upon tumour necrosis factor alpha-activation of PEC-A cells. However, the magnitude of this change induced by inflammatory conditions could be insufficient to affect cell surface alpha1,3 galactosylation. PMID- 11874462 TI - Identification of the 19S regulatory particle subunits from the rice 26S proteasome. AB - The 26S proteasome, a protein complex consisting of a 20S proteasome and a pair of 19S regulatory particles (RP), is involved in ATP-dependent proteolysis in eukaryotes. In yeast, the RP contains six different ATPase subunits and, at least, 11 non-ATPase subunits. In this study, we identified the rice homologs of yeast RP subunit genes from the rice expressed sequence tag (EST) library. The complete nucleotide sequences of the homologs for five ATPase subunits, OsRpt1, OsRpt2, OsRpt4, OsRpt5 and OsRpt6, and five non-ATPase subunits, OsRpn7, OsRpn8, OsRpn10, OsRpn11 and OsRpn12, and the partial sequences of one ATPase subunit, OsRpt3, and six non-ATPase subunits, OsRpn1, OsRpn2, OsRpn3, OsRpn5, OsRpn6 and OsRpn9, were determined. Gene homologs of four ATPase subunits, OsRpt1, OsRpt2, OsRpt4 and OsRpt5, and three non- ATPase subunits, OsRpn1, OsRpn2 and OsRpn9, were found to be encoded by duplicated genes. The rice RP was purified by immunoaffinity chromatography with a Protein A column immobilized antibody against rice 20S proteasome, and the subunit composition was determined. The homologs obtained from the rice EST library were identified as genes encoding subunits of RP purified from rice, including the both products of duplicated genes by using electrospray ionization quadrupole time-of-flight mass spectrometry. Post-translational modifications and processing in rice RP subunits were also identified. Various types of RP complex with different subunit compositions are present in rice cells, suggesting the multiple functions of rice proteasome. PMID- 11874463 TI - Formation of aberrant phosphotau fibrillar polymers in neural cultured cells. AB - Here we show, for the first time, the in vitro formation of filamentous aggregates of phosphorylated tau protein in SH-SY5Y human neuroblastoma cells. The formation of such aberrant aggregates, similar to those occurring in vivo in Alzheimer's disease and other tauopathies, requires okadaic acid, a phosphatase inhibitor, to increase the level of phosphorylated tau, and hydroxynonenal, a product of oxidative stress that selectively adducts and modifies phosphorylated tau. Our findings suggest that both phosphorylation and oxidative modification are required for tau filament formation. Importantly, the in vitro formation of intracellular tau aggregates could be used as a model of tau polymerization and facilitate the development of novel therapeutic approaches. PMID- 11874464 TI - Functional epitope of common gamma chain for interleukin-4 binding. AB - Interleukin 4 (IL-4) can act on target cells through an IL-4 receptor complex consisting of the IL-4 receptor alpha chain and the common gamma chain (gamma(c)). An IL-4 epitope for gamma(c) binding has previously been identified. In this study, the gamma(c) residues involved in IL-4 binding were defined by alanine-scanning mutational analysis. The epitope comprises gamma(c) residues I100, L102, and Y103 on loop EF1 together with L208 on loop FG2 as the major binding determinants. These predominantly hydrophobic determinants interact with the hydrophobic IL-4 epitope composed of residues I11, N15, and Y124. Double mutant cycle analysis revealed co-operative interaction between gamma(c) and IL-4 side chains. Several gamma(c) residues involved in IL-4 binding have been previously shown to be mutated in X-linked severe combined immunodeficiency. The importance of these binding residues for gamma(c) function is discussed. These results provide a basis for elucidating the molecular recognition mechanism in the IL-4 receptor system and a paradigm for other gamma(c)-dependent cytokine receptor systems. PMID- 11874465 TI - Variations in receptor site-3 on rat brain and insect sodium channels highlighted by binding of a funnel-web spider delta-atracotoxin. AB - Delta-atracotoxins (delta-ACTXs) from Australian funnel-web spiders differ structurally from scorpion alpha-toxins (Sc(alpha)Tx) but similarly slow sodium current inactivation and compete for their binding to sodium channels at receptor site-3. Characterization of the binding of 125I-labelled delta-ACTX-Hv1a to various sodium channels reveals a decrease in affinity for depolarized (0 mV; Kd=6.5 +/- 1.4 nm) vs.polarized (-55 mV; Kd=0.6 +/- 0.2 nm) rat brain synaptosomes. The increased Kd under depolarized conditions correlates with a 4.3 fold reduction in the association rate and a 1.8-increase in the dissociation rate. In comparison, Sc(alpha)Tx binding affinity decreased 33-fold under depolarized conditions due to a 48-fold reduction in the association rate. The binding of 125I-labelled delta-ACTX-Hv1a to rat brain synaptosomes is inhibited competitively by classical Sc(alpha)Txs and allosterically by brevetoxin-1, similar to Sc(alpha)Tx binding. However, in contrast with classical Sc(alpha)Txs, 125I-labelled delta-ACTX-Hv1a binds with high affinity to cockroach Na+ channels (Kd=0.42 +/- 0.1 nm) and is displaced by the Sc(alpha)Tx, Lqh(alpha)IT, a well defined ligand of insect sodium channel receptor site-3. However, delta-ACTX-Hv1a exhibits a surprisingly low binding affinity to locust sodium channels. Thus, unlike Sc(alpha)Txs, which are capable of differentiating between mammalian and insect sodium channels, delta-ACTXs differentiate between various insect sodium channels but bind with similar high affinity to rat brain and cockroach channels. Structural comparison of delta-ACTX-Hv1a to Sc(alpha)Txs suggests a similar putative bioactive surface but a 'slimmer' overall shape of the spider toxin. A slimmer shape may ease the interaction with the cockroach and mammalian receptor site-3 and facilitate its association with different conformations of the rat brain receptor, correlated with closed/open and slow-inactivated channel states. PMID- 11874466 TI - Regulation of RAS in human platelets. Evidence that activation of RAS is not sufficient to lead to ERK1-2 phosphorylation. AB - In this study, we show that the G protein-coupled receptor agonist thrombin, the glycoprotein VI agonist convulxin, and the cytokine receptor Mpl agonist thrombopoietin (TPO) are able to induce activation of RAS in human platelets. Recruitment of GRB2 by tyrosine-phosphorylated proteins in response to TPO and convulxin but not by thrombin occurred with a similar time-course to RAS activation, consistent with a causal relationship. On the other hand, activation of ERK2 by thrombin and convulxin is delayed and also inhibited by the protein kinase C inhibitor Ro-31 8220, whereas RAS activation is unaffected. Further evidence for differential regulation of RAS and ERK is provided by the observations that TPO, which activates RAS but not protein kinase C, does not activate ERK, and that the inhibitor of SRC kinases PP1 inhibits activation of RAS but not ERK2 in response to thrombin. Our results demonstrate that activation of RAS is not necessarily coupled to ERK in human platelets. PMID- 11874467 TI - Lectin-sugar interaction. Calculated versus experimental binding energies. AB - Although a steadily increasing number of protein--ligand docking experiments have been performed successfully, there are only few studies concerning protein--sugar interactions. In this study, we investigate the interaction of wheat germ agglutinin (WGA) with N-acetylglucosamine and a number of its derivatives and predict the binding free energies using flexible docking techniques. To assess the quality of our predictions, we also determined those binding free energies experimentally in cell-binding studies. The predicted binding site, ligand orientation, and details of the binding mode are in perfect agreement with the known crystal structure of WGA with a sialoglycopeptide. Furthermore, we obtained an excellent linear correlation of our predicted binding free energies with both our own data and experimental data from the literature [Monsigny, M., Roche, A.C., Sene, C., Maget Dana, R. & Delmotte, F. (1980) Eur. J. Biochem. 104, 147 153.]. In both cases, predicted energies were within 1.0 kJ x mol(-1) of the experimental value. These results illustrate the usefulness of docking-based methods for the qualitative and quantitative prediction of protein--carbohydrate interactions. The insights gained from such theoretical studies may be used to complement the results from the still scarce crystal structures. PMID- 11874468 TI - Holliday junction binding and processing by the RuvA protein of Mycoplasma pneumoniae. AB - The RuvA, RuvB and RuvC proteins of Escherichia coli act together to process Holliday junctions formed during recombination and DNA repair. RuvA has a well defined DNA binding surface that is sculptured specifically to accommodate a Holliday junction and allow subsequent loading of RuvB and RuvC. A negatively charged pin projecting from the centre limits binding of linear duplex DNA. The amino-acid sequences forming the pin are highly conserved. However, in certain Mycoplasma and Ureaplasma species the structure is extended by four amino acids and two acidic residues forming a crucial charge barrier are missing. We investigated the significance of these differences by analysing RuvA from Mycoplasma pneumoniae. Gel retardation and surface plasmon resonance assays revealed that this protein binds Holliday junctions and other branched DNA structures in a manner similar to E. coli RuvA. Significantly, it binds duplex DNA more readily. However it does not support branch migration mediated by E. coli RuvB and when bound to junction DNA is unable to provide a platform for stable binding of E. coli RuvC. It also fails to restore radiation resistance to an E. coli ruvA mutant. The data presented suggest that the modified pin region retains the ability to promote junction-specific DNA binding, but acts as a physical obstacle to linear duplex DNA rather than as a charge barrier. They also indicate that such an obstacle may interfere with the binding of a resolvase. Mycoplasma species may therefore process Holliday junctions via uncoupled branch migration and resolution reactions. PMID- 11874469 TI - Functional studies of the Synechocystis phycobilisomes organization by high performance liquid chromatography on line with a mass spectrometer. AB - This study was designed to yield data on the supramolecular organization of the phycobilisome apparatus from Synechocystis, and the possible effects of environmental stress on this arrangement. Phycobilisomes were dissociated in a low ionic strength solution and a quantitative estimation of the protein components present in each subcomplex was obtained using liquid chromatography coupled on-line with a mass spectrometer equipped with an electrospray ion source (ESI-MS). An advantage of this approach is that information can be collected on the initial events, which take place as this organism adapts to environmental changes. Ultracentrifugation of whole phycobilisomes revealed five subcomplexes; the lightest contained four linker proteins plus free phycocyanin, the second the core complex, while the last three bands contained the rod complexes. Four linkers were found in band 1 with higher molecular masses than those expected from the DNA sequence, indicating that they also contain linked chemical groups. UV-B irradiation specifically destroyed the beta-phycocyanin and one rod linker, which resulted in the disintegration of the rod complexes. The two bilins present in beta-phycocyanin give a greater contribution to the UV absorption than the single bilin of the other bilinproteins and probably react with atmospheric oxygen forming toxic radicals. The protein backbone is, in fact, protected from damage in anaerobic conditions and in the presence of radical scavengers. Cells grown in sulfur- and nitrogen-deficient medium contained significantly reduced levels of beta-phycocyanin and one rod linker. PMID- 11874470 TI - Irregular spiking in free calcium concentration in single, human platelets. Regulation by modulation of the inositol trisphosphate receptors. AB - Fluorescence ratio imaging indicates that immobilized, aspirin-treated platelets, loaded with Fura-2, respond to inositol 1,4,5-trisphosphate- (InsP3)-generating agonists such as thrombin by high-frequency, irregular rises in cytosolic [Ca2+]i with spikes that vary in peak level and peak-to-peak interval. This differs from the regular [Ca2+]i oscillations observed in other, larger cells. We found that the thiol-reactive compounds thimerosal (10 microm) and U73122 (10 microm) evoked similar irregular Ca2+ responses in platelets, but in this case in the absence of InsP3 generation. Thrombin-induced spiking was acutely abolished by inhibiting phospholipase C or elevating intracellular cAMP levels, while spiking with sulfhydryl reagents was only partially blocked by cAMP elevation. Confocal laser scanning microscopy using fluo-3-loaded platelets indicated that, with all agonists or conditions, the irregular spikes were almost instantaneously raised in various regions within a single platelet. When using saponin-permeabilized platelets, we found that InsP3-induced Ca2+ release from stores was stimulated by modest Ca2+ concentrations, pointing to a mechanism of InsP3-dependent Ca2+ induced Ca2+ release (CICR). This process was completely inhibitable by heparin. The Ca2+ release by InsP3, but not the CICR sensor, was negatively regulated by cAMP elevation. Thimerosal treatment did not release Ca2+ from intracellular stores, but markedly potentiated the stimulatory effect of InsP3. In contrast, U73122 caused a heparin/cAMP-insensitive Ca2+ leak from stores that differed from those used by InsP3. Taken together, these results demonstrate that InsP3 receptor channels play a crucial role in the irregular, spiking Ca2+ signal of intact platelets, even when induced by agents such as thimerosal or U73122 which do not stimulate InsP3 formation. The irregular Ca2+ release events appear to be subjected to extensive regulation by: (a) InsP3 level, (b) the potentiating effect of elevated Ca2+ on InsP3 action via CICR, (c) InsP3 channel sensitization by sulfhydryl (thimerosal) modification, (d) InsP3 channel-independent Ca2+ leak with U73122, and (e) down-regulation via cAMP elevation. The observation that individual Ca2+ peaks were generated in various parts of a platelet at similar intervals and amplitudes points to effective cooperation of the various stores in the Ca2+-release process. PMID- 11874471 TI - Mydj2 as a potent partner of hsc70 in mammalian cells. AB - Dj2 is a member of the DnaJ family of proteins, which regulate the chaperoning function of the hsp70s. We isolated a monkey cDNA dj2 clone corresponding to the large mRNA species encoded by the gene. This mRNA differs from the small mRNA produced by the same gene in that it contains a long 3' untranslated region. Both messages were found to be equally stable and to produce the same protein, which is susceptible to farnesylation. Studies in mouse tissues and various cell lines revealed that these messages and their products are differentially expressed. Surprisingly, we found that only the nonfarnesylated form of dj2 is capable of translocating to the cell nucleus, especially after heat shock. Finally, based on protein interaction studies, our results indicate that dj2 is a specific partner for hsc70 and not for hsp70. PMID- 11874472 TI - Ubiquitination of soluble and membrane-bound tyrosine hydroxylase and degradation of the soluble form. AB - Tyrosine hydroxylase (TH) demonstrates by two-dimensional electrophoresis a microheterogeneity both as a soluble recombinant human TH (hTH1) and as a membrane-bound bovine TH (bTHmem). Part of the heterogeneity is likely due to deamidation of labile asparagine residues. Wild-type (wt)-hTH1 was found to be a substrate for the ubiquitin (Ub) conjugating enzyme system in a reconstituted in vitro system. When wt-hTH1 was expressed in a coupled transcription-translation TnT(R)-T7 reticulolysate system 35S-labelled polypeptides of the expected molecular mass of native enzyme as well as both higher and lower molecular mass forms were observed. The amount of high-molecular-mass forms increased by time and was enhanced in the presence of Ub and clasto-lactacystin beta-lactone. In pulse-chase experiments the amount of full-length hTH1 decreased by first-order kinetics with a half-time of 7.4 h and 2.1 h in the absence and presence of an ATP-regenerating system, respectively. The ATP-dependent degradation was inhibited by clasto-lactacystin beta-lactone. Our findings support the conclusion that hTH1 is ubiquitinated and at least partially degraded by the proteasomes in the reticulocyte lysate system. Finally, it is shown that the integral TH of the bovine adrenal chromaffin granule membrane (bTHmem) is ubiquitinated, most likely monoubiquitinated. Additional Ub-conjugates of this membrane, detected by Western blot analysis, have not yet been identified. PMID- 11874473 TI - The expression of glutathione reductase in the male reproductive system of rats supports the enzymatic basis of glutathione function in spermatogenesis. AB - Glutathione reductase (GR) recycles oxidized glutathione (GSSG) by converting it to the reduced form (GSH) using an NADPH as the electron source. The function of GR in the male genital tract of the rat was examined by measuring its enzymatic activity and examining the gene expression and localization of the protein. Levels of GR activity, the protein, and the corresponding mRNA were the highest in epididymis among testes, vas deferens, seminal vesicle, and prostate gland. The localization of GR, as evidenced by immunohistochemical techniques, reveals that it exists at high levels in the epithelia of the genital tract. In testis, GR is mainly localized in Sertoli cells. The enzymatic activity and protein expression of GR in primary cultured testicular cells confirmed its predominant expression in Sertoli cells. Intracellular GSH levels, expressed as mol per mg protein, was higher in spermatogenic cells than in Sertoli cells. As a result of these findings, the effects of buthionine sulfoximine (BSO), an inhibitor for GSH synthesis, and 1,3-bis(2-chlorethyl)-1-nitrosourea (BCNU), an inhibitor for GR, on cultured testicular cells were examined. Sertoli cells were prone to die as the result of BCNU, but not BSO treatment, although intracellular levels of GSH declined more severely with BSO treatment. Spermatogenic cells were less sensitive to these agents than Sertoli cells, which indicates that the contribution of these enzymes is less significant in spermatogenic cells. The results herein suggest that the GR system in Sertoli cells is involved in the supplementation of GSH to spermatogenic cells in which high levels of cysteine are required for protamine synthesis. In turn, the genital tract, the epithelia of which are rich in GR, functions in an antioxidative manner to protect sulfhydryl groups and unsaturated fatty acids in spermatozoa from oxidation during the maturation process and storage. PMID- 11874474 TI - Introducing Wilson disease mutations into the zinc-transporting P-type ATPase of Escherichia coli. The mutation P634L in the 'hinge' motif (GDGXNDXP) perturbs the formation of the E2P state. AB - ZntA, a bacterial zinc-transporting P-type ATPase, is homologous to two human ATPases mutated in Menkes and Wilson diseases. To explore the roles of the bacterial ATPase residues homologous to those involved in the human diseases, we have introduced several point mutations into ZntA. The mutants P401L, D628A and P634L correspond to the Wilson disease mutations P992L, D1267A and P1273L, respectively. The mutations D628A and P634L are located in the C-terminal part of the phosphorylation domain in the so-called hinge motif conserved in all P-type ATPases. P401L resides near the N-terminal portion of the phosphorylation domain whereas the mutations H475Q and P476L affect the heavy metal ATPase-specific HP motif in the nucleotide binding domain. All mutants show reduced ATPase activity corresponding 0-37% of the wild-type activity. The mutants P401L, H475Q and P476L are poorly phosphorylated by both ATP and P(i). Their dephosphorylation rates are slow. The D628A mutant is inactive and cannot be phosphorylated at all. In contrast, the mutant P634L six residues apart in the same domain shows normal phosphorylation by ATP. However, phosphorylation by P(i) is almost absent. In the absence of added ADP the P634L mutant dephosphorylates much more slowly than the wild-type, whereas in the presence of ADP the dephosphorylation rate is faster than that of the wild-type. We conclude that the mutation P634L affects the conversion between the states E1P and E2P so that the mutant favors the E1 or E1P state. PMID- 11874475 TI - Mast cell-fibroblast interactions: human mast cells as source and inducers of fibroblast and epithelial growth factors. AB - As mast cells have been implicated in cutaneous repair processes, we have examined the ability of human mast cells to produce important epithelial and fibroblast growth factors or to stimulate the production of such factors in dermal fibroblasts. Isolated, highly purified human dermal mast cells and human leukemic mast cells were examined for mRNA and partly also for protein expression of these molecules as such or after preincubation with interleukin-4, stem cell factor, or with phorbol myristate acetate. In addition, mast cells were studied for their ability to induce fibroblast growth factor 2 and fibroblast growth factor 7 secretion from dermal fibroblasts. Both dermal and leukemic mast cells expressed fibroblast growth factor 2, fibroblast growth factor 7, and heparin binding epidermal growth factor, but not hepatocyte growth factor at mRNA level, and dermal mast cells expressed fibroblast growth factor 10 in addition. At protein level, spontaneous fibroblast growth factor 2 secretion was noted that was markedly enhanced by phorbol myristate acetate, whereas no fibroblast growth factor 7 protein was detected under these conditions. Instead, human mast cell-1 supernatants induced enhanced fibroblast growth factor 7 secretion from dermal fibroblasts, with phorbol-myristate-acetate-stimulated supernatants being more effective. This effect could be reproduced with histamine and was H1-receptor mediated. Tryptase was ineffective but stimulated instead fibroblast growth factor 2 secretion from fibroblasts. These data demonstrate for the first time the ability of mast cells to express and/or secrete several growth factors of the fibroblast growth factor family as well as heparin-binding epidermal growth factor directly or indirectly via stimulation of fibroblasts, underlining the potentially pivotal role of these cells during human tissue repair and homeostasis. PMID- 11874476 TI - Nerve growth factor, neuropeptides, and mast cells in ultraviolet-B-induced systemic suppression of contact hypersensitivity responses in mice. AB - The induction of systemic immunosuppression following ultraviolet B radiation exposure has been linked with the release of inflammatory and immunomodulatory mediators by cells of the epidermis and dermis. Nerve growth factor has not previously been linked with ultraviolet-B-induced immunosuppressive effects. Nerve growth factor antibodies abrogated ultraviolet-B-induced systemic suppression of contact hypersensitivity responses in BALB/C mice. Subcutaneous injection of nerve growth factor (20 microg per mouse) into dorsal skin 5 d before hapten sensitization on ventral skin suppressed contact hypersensitivity responses in mast-cell-replete but not Wf/Wf mast-cell-depleted mice. Nerve growth factor injected 24 h prior to challenge was not able to suppress the efferent phase of the contact hypersensitivity response. Subcutaneous injection of nerve growth factor (20 microg per mouse) did not suppress contact hypersensitivity responses in capsaicin-pretreated (neuropeptide-depleted) BALB/c mice, and thus sensory c-fibers are necessary for nerve-growth-factor-mediated systemic suppression of contact hypersensitivity responses. Increased concentrations of nerve growth factor within epidermal keratinocytes 8 h after ultraviolet B irradiation were confirmed immunohistochemically. These findings support a role for keratinocyte-derived nerve growth factor via its action on sensory c-fibers, and subsequent release of neuropeptides to mediate mast cell degranulation in systemic suppression of contact hypersensitivity responses in mice following ultraviolet B exposure. PMID- 11874477 TI - Connective tissue growth factor: expression in human skin in vivo and inhibition by ultraviolet irradiation. AB - Connective tissue growth factor, which is induced by transforming growth factor beta, has been reported to mediate the stimulatory actions of transforming growth factor beta on type I procollagen synthesis. Connective tissue growth factor is expressed in fibrotic disease such as scleroderma, where it is believed to promote abnormal deposition of collagen. Connective tissue growth factor expression has not been described in normal human skin or cultured skin cells, however. We report here that connective tissue growth factor mRNA is constitutively expressed in normal human skin. In situ hybridization demonstrated that connective tissue growth factor mRNA was expressed in keratinocytes throughout the epidermis and in dermal cells. Quantitative real-time reverse transcription polymerase chain reaction revealed that the level of connective tissue growth factor mRNA in the epidermis and dermis of normal human skin was comparable to the level of housekeeping gene 36B4. Ultraviolet irradiation (2 minimal erythema dose, UVB/A2 source) reduced connective tissue growth factor mRNA expression throughout the epidermis and dermis in normal human skin in vivo. Connective tissue growth factor mRNA was reduced (30%) within 4 h post ultraviolet irradiation, and remained reduced (50%) 8-24 h post ultraviolet. Connective tissue growth factor mRNA and protein were also constitutively highly expressed in normal cultured human skin keratinocytes and fibroblasts. Ultraviolet irradiation of cultured normal human skin fibroblasts resulted in a time-dependent inhibition of connective tissue growth factor mRNA expression. At 24 h post ultraviolet, connective tissue growth factor mRNA expression was reduced 80%. Transforming growth factor beta1 rapidly induced connective tissue growth factor mRNA levels (5-fold within 4 h) in skin fibroblasts, but not keratinocytes, and this induction was attenuated 80% by ultraviolet irradiation. Electrophoretic mobility shift assays demonstrated that ultraviolet irradiation reduced protein binding to the transforming growth factor beta/Smad responsiveness elements in the connective tissue growth factor gene promoter, in human skin in vivo and human skin fibroblasts. Constitutive expression of connective tissue growth factor in normal human skin suggests that it is a physiologic regulator of procollagen synthesis. Ultraviolet reduction of connective tissue growth factor expression may contribute to reduced procollagen synthesis observed in ultraviolet-irradiated normal human skin and human skin fibroblasts. PMID- 11874478 TI - Alterations in fibroblast alpha1beta1 integrin collagen receptor expression in keloids and hypertrophic scars. AB - Keloids and hypertrophic scars are significant symptomatic clinical problems characterized by excess collagen. Although extensive research has focused on fibroblasts and collagen turnover in these aberrant scars, little work has been done on the expression of integrins (cell membrane structures that link cells to extracellular matrix) within these lesions. Integrin-mediated regulation of collagen synthesis has previously been observed in explanted fibroblasts from normal and fibrotic dermis, and integrin alpha1 knockout mice maintain increased collagen synthesis consistent with a role for alpha1beta1 in providing negative feedback on collagen synthesis. These findings suggested the need to evaluate integrin roles in keloids and hypertrophic scars. In this study we examined integrin expression in keloids (n = 11), hypertrophic scars (n = 5), radiation ulcers (n = 2), and normal skin specimens (n = 8). We used a novel approach to analysis by isolating dermal fibroblasts directly from tissue (without explant culture) and determining surface integrin expression by flow cytometry. We found that keloids and hypertrophic scars have marked alterations in fibroblast integrin expression and contain several distinct populations of fibroblasts. One of these populations expresses high levels of alpha1 integrin, and the proportion of these cells is higher in keloids (63% +/- 3.6% SEM) and hypertrophic scars (45% +/- 2.7% SEM) than in normal skin tissues (28% +/- 4.7% SEM). The different populations of fibroblasts defined by integrin expression merge, however, when the cells are serially cultured, suggesting that there may be aspects of the dermal microenvironment that maintain the integrin phenotypic heterogeneity in dermal fibroblasts. PMID- 11874479 TI - Creatine kinase and creatine transporter in normal, wounded, and diseased skin. AB - Skin comprises many cell types that are characterized by high biosynthetic activity and increased energy turnover. The creatine kinase system, consisting of creatine kinase isoenzymes and creatine transporter, is known to be important to support the high energy demands in such cells. We analyzed the presence and the localization of these proteins in murine and human skin under healthy and pathologic conditions, using immunoblotting and confocal immunohistochemistry with our recently developed specific antibodies. In murine skin, we found high amounts of brain-type cytosolic creatine kinase coexpressed with lower amounts of ubiquitous mitochondrial creatine kinase, both mainly localized in suprabasal layers of the epidermis, different cell types of hair follicles, sebaceous glands, and the subcutaneous panniculus carnosus muscle. With exception of sebaceous glands, these cells were also expressing creatine transporter. Muscle type cytosolic creatine kinase and sarcomeric mitochondrial creatine kinase were restricted to panniculus carnosus. Immediately after wounding of murine skin, brain-type cytosolic creatine kinase and a creatine transporter-subspecies were transiently upregulated about 3-fold as seen in immunoblots, whereas the amount of ubiquitous mitochondrial creatine kinase increased during days 10-15 after wounding. Healthy and psoriatic human skin showed a similar coexpression pattern of brain-type cytosolic creatine kinase, ubiquitous mitochondrial creatine kinase, and creatine transporter in this pilot study, with creatine transporter species being upregulated in psoriasis. PMID- 11874480 TI - Enhanced cutaneous inflammatory reactions to Aspergillus fumigatus in a murine model of chronic granulomatous disease. AB - Chronic granulomatous disease is the manifestation of genetic defects of the leukocyte NADPH oxidase resulting in the absence of a respiratory burst. Patients with chronic granulomatous disease can develop chronic granulomas in many locations of the body, including the skin. Using an established murine model of X linked chronic granulomatous disease (X-CGD) created by homologous recombinant disruption of the gene encoding the gp91phox component of the NADPH oxidase, in this study we examined cutaneous reactivity to sterile Aspergillus fumigatus hyphae. Injection of Aspergillus fumigatus into the dorsal ears of X-CGD mice resulted in an enhanced inflammatory response by 24 h, consisting of neutrophils, which developed into suppurative granulomas by 10 d. Intradermal injection of Aspergillus fumigatus into wild-type mice only resulted in a transient inflammatory response that resolved by 10 d. Injection of Aspergillus fumigatus into female carrier mice resulted in an acute inflammatory response that was similar to that of wild-type mice, but, at higher doses of Aspergillus fumigatus, many carriers subsequently developed granulomatous lesions that were qualitatively similar but smaller than those seen in X-CGD mice by 30 d. Consistent with the ability of X-CGD mice to mount an enhanced neutrophil-rich inflammatory response to Aspergillus fumigatus, significant levels of the potent neutrophil activator/chemoattractant leukotriene B4 were measured by mass spectrometry in skin biopsies at 24 and 72 h. In contrast to the exaggerated inflammatory response to intradermal Aspergillus fumigatus in X-CGD mice compared to their wild-type counterparts, similar levels of inflammation were seen in a model of delayed-type hypersensitivity using 2,4-dinitrofluorobenzene. This study represents the first report of a cutaneous granuloma model in mice with X-CGD, which may also prove useful as a functional test to evaluate the efficacy of gene therapy protocols being developed for chronic granulomatous disease. PMID- 11874481 TI - Altered water barrier function in epidermal-type fatty acid binding protein deficient mice. AB - We have generated mutant mice for epidermal-type fatty acid binding protein by the gene targeting technique and examined the phenotype in detail. Despite a lack in the expression of epidermal-type fatty acid binding protein mRNA and its protein in the skin and other tissues of the mutant mice, the animals appeared normal in gross and histologic examination. Northern blot analysis of other fatty acid binding proteins revealed a distinct elevated gene expression of heart-type fatty acid binding protein in the skin of the homozygous mice. In analyses of the skin, no differences were observed in contents of major fatty acids, electron microscopic appearance as well as inflammatory responses in ear skin between the mutant and wild-type mice. Basal transepidermal water loss of homozygous mice was lower than that of the wild mice. When acetone was applied to the skin for disruption of the water permeability barrier, recovery in transepidermal water loss was delayed, although maximum transepidermal water loss upon acetone treatment was similar between homozygous and wild-type mice in terms of size and time course. The molecular mechanism by which epidermal-type fatty acid binding protein contributes to the water barrier function of the skin remains to be elucidated. PMID- 11874483 TI - Epidermal overexpression of stratum corneum chymotryptic enzyme in mice: a model for chronic itchy dermatitis. AB - Identification of tissue-specific mechanisms involved in the pathophysiology of inflammatory skin diseases could offer new possibilities to develop effective therapies with fewer systemic effects. The serine protease stratum corneum chymotryptic enzyme is preferentially expressed in cornifying epithelia. We have previously reported on increased expression of the stratum corneum chymotryptic enzyme in psoriasis. Here is reported an increased epidermal expression of stratum corneum chymotryptic enzyme also found in chronic lesions of atopic dermatitis. Transgenic mice expressing human stratum corneum chymotryptic enzyme in suprabasal epidermal keratinocytes were found to develop pathologic skin changes with increased epidermal thickness, hyperkeratosis, dermal inflammation, and severe pruritus. The results suggest that stratum corneum chymotryptic enzyme may be involved in the pathogenesis of inflammatory skin diseases, and that stratum corneum chymotryptic enzyme and related enzymes should be evaluated as potential targets for new therapies. PMID- 11874482 TI - Elevated stratum corneum hydrolytic activity in Netherton syndrome suggests an inhibitory regulation of desquamation by SPINK5-derived peptides. AB - Netherton syndrome is a congenital ichthyosis associated with erythroderma, hair shaft defects, and atopic features. The mutations of the secretory serine protease inhibitor Kazal-type 5 gene have been identified in Netherton syndrome patients; however, the actual physiologic substrates of the serine protease inhibitor Kazal-type 5 proprotein are unknown, and how the genetic defects cause characteristic skin phenotype remains uncertain. Here, we describe the serine protease inhibitor Kazal-type 5 gene mutations, including two novel non-sense mutations, and genotype-phenotype correlation in three Netherton syndrome patients in two unrelated Japanese families. Furthermore, based on the reappraisal of the structure of the serine protease inhibitor Kazal-type 5 proprotein, demonstration of the presence of carboxypeptidase in normal keratinocytes, and the observation of mRNA localization of the serine protease inhibitor Kazal-type 5 transcripts in the uppermost epidermis as well as pilosebaceous units, we propose a hypothetical model of proteolytic processing of the serine protease inhibitor Kazal-type 5 proprotein in the epidermis and inhibitory regulation of corneocyte desquamation by a set of serine protease inhibitor Kazal-type 5-derived peptides. This hypothesis is supported by the marked increase of trypsin-like hydrolytic activity demonstrated in stratum corneum samples from our Netherton syndrome patients. The findings in this study suggest that the defective inhibitory regulation of desquamation due to the serine protease inhibitor Kazal-type 5 gene mutations may cause over-desquamation of corneocytes in Netherton syndrome, leading to severe skin permeability barrier dysfunction. PMID- 11874484 TI - Cord blood CD34+ cells differentiate into dermal dendritic cells in co-culture with cutaneous fibroblasts or stromal cells. AB - The skin is a unique organ that contains two different subsets of dendritic cells, i.e., Langerhans cells and dermal dendritic cells. Our hypothesis is that cutaneous fibroblasts may affect the development of these dendritic cells. We cocultured cord blood CD34+ hematopoietic progenitor cells with several human cutaneous fibroblast cell lines without any exogenous cytokines for 3 wk. In this culture, hematopoietic progenitor cells increased in number from 20.1 +/- 2.4 times, and produced aggregates of cells with dendritic processes. They were composed of 54.9 +/- 3.2% HLA-DR+ CD14+ CD1a-- cells and 13.8 +/- 3.6% HLA-DR+ CD1a+ cells, which also expressed CD11b and CD11c. There were significant numbers of factor XIIIa+ cells in the culture, whereas no Lag+ or E-cadherin+ cells were detected, and they were potent stimulators in allogeneic T cell activation. There was a significant difference in the ability to induce CD1a+ cells among different human cutaneous fibroblast cell lines. These CD1a+ cells lacked the expression of CD80, CD86, or CD83. In addition, half of them still expressed CD14. When these dendritic cells were cultured with tumor necrosis factor-alpha, however, they became mature dendritic cells with augmented expression of CD86 and CD83 and with increased allogeneic T cell stimulation. The subsequent experiment using a dividing chamber, enzyme-linked immunosorbent assay for granulocyte-macrophage colony-stimulating factor and macrophage colony-stimulating factor, and the blocking studies with antibodies for these cytokines suggested that both the presence of direct contact between hematopoietic progenitor cells and human cutaneous fibroblast cell lines and macrophage colony-stimulating factor produced by human cutaneous fibroblast cell lines are required for their maximum growth and differentiation into CD1a+ dendritic cells, whereas macrophage colony stimulating factor was solely responsible for their differentiation. These data suggest that cutaneous fibroblasts support the differentiation of dermal dendritic cells in addition to that of monocytes from hematopoietic progenitor cells by their direct contact with hematopoietic progenitor cells and by their macrophage colony-stimulating factor production. PMID- 11874485 TI - Halofuginone, an inhibitor of type-I collagen synthesis and skin sclerosis, blocks transforming-growth-factor-beta-mediated Smad3 activation in fibroblasts. AB - Halofuginone is a drug that has been shown to have an antifibrotic property in vitro and in vivo. Whereas halofuginone shows promise as a therapeutic agent for a variety of diseases including scleroderma, liver cirrhosis, cystic fibrosis, and certain types of cancer, the mechanism of action remains unknown. Using the tight skin mouse (TSK) model for scleroderma, we evaluated the ability of halofuginone to inhibit spontaneous development of dermal fibrosis. We found that administration of a low dose of halofuginone both in adult and newborn animals for 60 d prevented the development of cutaneous hyperplasia (dermal fibrosis). In vitro halofuginone was found to reduce the amount of collagen synthesized by fibroblasts. This effect was due to a reduction in the promoter activity of the type-I collagen genes as treatment of fibroblast cultures with 10(-8) M halofuginone reduced the level of alpha2(I) collagen message detectible by northern blot and greatly reduced the activity of a reporter construct under control of the -3200 to +54 bp alpha2(I) collagen promoter. In addition, analysis of transforming growth factor beta signaling pathways in fibroblasts revealed that halofuginone inhibited transforming-growth-factor-beta-induced upregulation of collagen protein and activity of the alpha2(I) collagen promoter. Further we found that halofuginone blocked the phosphorylation and subsequent activation of Smad3 after transforming growth factor beta stimulation. Apparently the inhibitory property was specific to Smad3 as there was no inhibitory effect on the activation of Smad2 after stimulation with transforming growth factor beta. Our results demonstrate that halofuginone is a specific inhibitor of type-I collagen synthesis and may elicit its effect via interference with the transforming growth factor beta signaling pathway. PMID- 11874486 TI - Transforming growth factor beta1 regulates melanocyte proliferation and differentiation in mouse neural crest cells via stem cell factor/KIT signaling. AB - Stem cell factor is essential to the migration and differentiation of melanocytes during embryogenesis based on the observation that mutations in either the stem cell factor gene, or its ligand, KIT, result in defects in coat pigmentation in mice. Stem cell factor is also required for the survival of melanocyte precursors while they are migrating towards the skin. Transforming growth factor beta1 has been implicated in the regulation of both cellular proliferation and differentiation. NCC-melb4, an immortal cloned cell line, was cloned from a mouse neural crest cell. NCC-melb4 cells provide a model to study the specific stage of differentiation and proliferation of melanocytes. They also express KIT as a melanoblast marker. Using the NCC-melb4 cell line, we investigated the effect of transforming growth factor beta1 on the differentiation and proliferation of immature melanocyte precursors. Immunohistochemically, NCC-melb4 cells showed transforming growth factor beta1 expression. The anti-transforming growth factor beta1 antibody inhibited the cell growth, and downregulated the KIT protein and mRNA expression. To investigate further the activation of autocrine transforming growth factor beta1, NCC-melb4 cells were incubated in nonexogenous transforming growth factor beta1 culture medium. KIT protein decreased with anti-transforming growth factor beta1 antibody concentration in a concentration-dependent manner. We concluded that in NCC-melb4 cells, transforming growth factor beta1 promotes melanocyte precursor proliferation in autocrine and/or paracrine regulation. We further investigated the influence of transforming growth factor beta1 in vitro using a neural crest cell primary culture system from wild-type mice. Anti transforming growth factor beta1 antibody decreased the number of KIT positive neural crest cell. In addition, the anti-transforming growth factor beta1 antibody supplied within the wild-type neural crest explants abolished the growth of the neural crest cell. These results indicate that transforming growth factor beta1 affect melanocyte precursor proliferation and differentiation in the presence of stem cell factor/KIT in an autocrine/paracrine manner. PMID- 11874487 TI - Advanced glycation end product-modified beta2-microglobulin is a component of amyloid fibrils of primary localized cutaneous nodular amyloidosis. AB - Primary localized cutaneous nodular amyloidosis is a rare form of cutaneous amyloidosis. Amyloid fibrils in primary localized cutaneous nodular amyloidosis have been reported to be originated from immunoglobulin light chains. Immunohistochemical studies on the lesional skins of four patients with primary localized cutaneous nodular amyloidosis demonstrated that amyloid deposits of all cases showed a positive reaction with the antibodies for beta2-microglobulin and advanced glycation end products as well as immunoglobulin light chain (kappa or lambda). No beta2-microglobulin and advanced glycation end product immunoreactivity was found in the amyloid deposits of other primary localized cutaneous amyloidosis (lichen amyloidosis and macular amyloidosis). Double immunofluorescence study of the lesional skin of primary localized cutaneous nodular amyloidosis showed that anti-kappa light chain, anti-beta2-microglobulin and anti-advanced glycation end product antibodies mostly reacted with the same area of amyloid deposit. Amyloid proteins were sequentially extracted with distilled water from one case of primary localized cutaneous nodular amyloidosis and recovered in the five water-soluble fractions (fractions I-V). Immunoblot assay of amyloid fibril proteins demonstrated that immunoreactive polypeptides with anti-kappa light chain antibody (29 kDa) and with anti-beta2-microglobulin antibody (12 kDa) were detected in fractions I-V, whereas immunoreactive polypeptide with anti-advanced glycation end product antibody (12 kDa) was detected exclusively in fractions III-V but not in fractions I and II. Two dimensional polyacrylamide gel electrophoresis revealed that 12 kDa polypeptide in fractions I and II was electrophoretically identical with authentic beta2 microglobulin and that beta2-microglobulin in fractions III-V was advanced glycation end product-modified beta2-microglobulin with more acidic pI value. These results indicate that beta2-microglobulin is another major component of amyloid fibrils in primary localized cutaneous nodular amyloidosis and that beta2 microglobulin in primary localized cutaneous nodular amyloidosis is partly subjected to the modification of advanced glycation end product. PMID- 11874488 TI - Cloning of hamster type XVII collagen cDNA, and pathogenesis of anti-type XVII collagen antibody and complement in hamster bullous pemphigoid. AB - Bullous pemphigoid is an inflammatory subepidermal blistering skin disease associated with an IgG autoimmune response to the type XVII collagen. The immunopathologic features of bullous pemphigoid can be reproduced in mice by the passive transfer of anti-type XVII collagen antibodies. In this model, it is thought that blister formation depends upon complement activation, neutrophil recruitment, and some proteolytic enzymes. In this study, we cloned hamster type XVII collagen cDNA, which contains a 4296 bp coding region and which is predicted to be a transmembrane protein with an extracellular collagenous domain, residing in type II orientation. Antipeptide antibodies (anti-1191 IgG) were obtained against a segment of hamster type XVII collagen homologous with the human type XVII collagen autoantibody-reactive site. The antipeptide antibodies were passively transferred to neonatal Syrian hamsters. The injected hamsters developed a microscopic subepidermal blister as seen previously in the mice. In order to test whether antigen-antibody complexes and complement initiate the subepidermal blister formation, we carried out experiments in vitro on condition that inflammatory cells were completely eliminated. Complement activation in sera was inhibited either by heating (at 56 degrees C for 30 min) or by preincubating with cobra venom factor. When the hamster skin was incubated with fresh anti-1191 antisera, separation of dermal-epidermal junction was observed. The anti-1191 IgG failed to induce C3 deposition and dermal-epidermal junction separation, however, if the anti-1191 IgG was added alone or complement activation in sera was inhibited. Under these conditions, IgG but not C3 was deposited on the basement membrane. These results strongly suggest that antigen-antibody complexes and complement initiate dermal-epidermal junction separation. PMID- 11874489 TI - Frequent abnormalities of the p15 and p16 genes in mycosis fungoides and sezary syndrome. AB - There are few data on the molecular pathogenesis of cutaneous T cell lymphomas. A recent allelotyping study by our group identified frequent allelic loss on 9p, 10q, and 17p including losses on 9p21 in 16% of patients with mycosis fungoides and 46% with Sezary syndrome. The P15 and P16 genes are intricately linked on 9p21 and can be inactivated in melanoma and non-Hodgkin's lymphoma. We have therefore studied 76 patients with either mycosis fungoides or Sezary syndrome for abnormalities of these genes. DNA samples were analyzed for loss of heterozygosity, homozygous deletion, intragenic mutations, and promoter methylation. In addition P15 and P16 protein expression was assessed. Microsatellite analysis was informative in 73 of 76 cases: allelic loss on 9p21 was identified in 18 patients (25%), including 12 of 57 with mycosis fungoides (21%) and six of 16 with Sezary syndrome (37%). Single strand conformation polymorphism analysis of the entire coding regions of both genes did not identify any mutations, although two polymorphisms were identified including C613A, which has not previously been described. P15 and P16 gene promoter methylation was found in 45% and 29% of patients, respectively. Furthermore aberrant P15 protein expression was detected in 85% of patients analyzed with P15 gene abnormalities and abnormal P16 expression in 59% with P16 gene abnormalities. These abnormalities were not dependent on cutaneous stage of disease. This study suggests that abnormalities of the P15 and P16 genes are common in both early and advanced stages of mycosis fungoides and Sezary syndrome and that these genes may be inactivated by allelic loss and aberrant promoter methylation. PMID- 11874490 TI - Evolution of melanocytic nevi on the faces and necks of adolescents: a 4 y longitudinal study. AB - All melanocytic nevi on the faces and necks of a cohort of students, initially aged 12-14 y, were mapped and photographed annually for 4 y. The features of each nevus were charted yearly noting changes in size and profile, and the appearance or disappearance of any nevi on a student's face and neck was recorded. Nevi were classified by size (small, < 2 mm; medium, 2-5 mm; large, > 5 mm), and by profile (flat, raised). Data from 20 adolescents selected randomly from the cohort for detailed analysis showed males had about twice as many nevi as females, but there was little difference between sexes in their patterns of nevus development. Approximately half the nevi were small in all years; under 5% were large. Over the 4 y of follow-up the proportion of flat nevi dropped from 70% to 57%, whereas nevus numbers increased by 47% in year 1, with smaller increases in older students. Most new or disappearing nevi were small and flat, although both incident and disappearing nevi could be larger and/or raised. Of the existing nevi that altered in the follow-up period, the tendency was towards an increase in size among raised but not among flat nevi; a lowering of profile among small nevi; and a raising of profile among larger nevi; but there were many exceptions to this pattern. Among several host factors examined, inability to tan after sun exposure was found to be significantly negatively associated with the propensity of nevi to change size over the study period. Overall our findings indicate that, contrary to conventional belief, there is a measurable turnover among melanocytic nevi even in early life. PMID- 11874491 TI - Bcl-2 and bcl-xL antisense oligonucleotides induce apoptosis in melanoma cells of different clinical stages. AB - Recent clinical studies have shown the promise of bcl-2 antisense therapy in patients with melanoma. To further demonstrate the importance of bcl-2 and validate the related antiapoptotic protein bcl-xL as targets for antisense therapy in melanoma, their implication as survival factors in melanoma cells of different clinical stages as well as in normal melanocytes was investigated. Primary cell cultures derived from 17 melanomas, the cell line A375, and normal melanocytes from healthy donors were treated with antisense oligonucleotides targeting either the bcl-xL mRNA or the bcl-2 and the bcl-xL mRNAs simultaneously. Bcl-2 and bcl-xL expression in cells was analyzed by real-time polymerase chain reaction and Western blotting. Cell viability was assessed in 3 (4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide and apoptosis assays. Bcl-2 expression was low in melanoma cells of stages I, II, and III, hardly detectable in A375 cells, but high in normal melanocytes. Bcl-xL expression was high in all cell types tested. As shown in A375 cells and the stage III melanoma cells 0513, both the bcl-xL monospecific oligonucleotide 4259 and the bcl-2/bcl xL bispecific oligonucleotide 4625 effectively reduced tumor cell viability by induction of apoptosis with IC50 values ranging from 200 to 350 nM. Oligonucleotide 4625 proved to be superior to 4259, as it significantly reduced the viability of cells from all melanoma stages. Both oligonucleotides reduced also the viability of normal melanocytes. Our data suggest that bcl-2 and bcl-xL are promising targets for antisense therapy of melanoma, and that the simultaneous downregulation of their expression may provide additional clinical benefit. PMID- 11874492 TI - Oxidative stress-independent depletion of epidermal vitamin A by UVA. AB - In hairless mice, epidermal vitamin A (retinol and retinyl esters) is strongly decreased following a single exposure to UVB. Here, using the same mouse model, we studied the effects of UVA on epidermal vitamin A content, lipid peroxidation, and CRBP-I expression, as well as the putative prevention of vitamin A depletion or lipid peroxidation by topical alpha-tocopherol. An acute exposure to UVA completely depleted epidermal vitamin A with EC50 of 0.25 and 0.5 J per cm2 for retinyl esters and retinol, respectively; these values were 0.1 J per cm2 for both retinoids under UVB exposure. CRBP-I expression was increased 2-fold 8 h following UVA exposure (10 J per cm2), and this increase persisted for at least 16 h. A single UVA exposure induced a concentration-dependent epidermal lipid peroxidation (EC50 = 3.5 J per cm2) giving rise to 55.4 +/- 4.2 nmol lipid peroxides per g at 20 J per cm2, whereas UVB, up to 1 J per cm2, did not increase the basal concentration of 6.7 +/- 0.9 nmol lipid peroxides per g. On the other hand, topical menadione induced a concentration-dependent lipid peroxidation, but did not affect vitamin A content. Pretreatment with alpha-tocopherol (i) did not inhibit UV-induced vitamin A depletion, (ii) completely inhibited the increased lipid peroxidation induced by UVA or menadione, and (iii) accelerated reconstitution of epidermal vitamin A after UVB but not UVA induced depletion. Thus acute UVA induced both epidermal vitamin A depletion and lipid peroxidation, UVB induced only vitamin A depletion, and menadione induced only a lipid peroxidation; topical alpha-tocopherol prevented lipid peroxidation but not vitamin A depletion. These observations indicate (i) that CRBP-I neither provides protection to UVB- and UVA-induced epidermal vitamin A depletion, nor interferes significantly with reconstitution, and (ii) that the UV-induced vitamin A depletion and lipid peroxidation in mouse epidermis are unrelated processes. UV light does not destroy epidermal vitamin A through an oxidative stress but probably by a photochemical reaction in which UV radiations at about 325 nm give the corresponding activation energy. PMID- 11874493 TI - 17beta-estradiol enhances vascular endothelial growth factor production and dihydrotestosterone antagonizes the enhancement via the regulation of adenylate cyclase in differentiated THP-1 cells. AB - We studied the in vitro effects of sex hormones on vascular endothelial growth factor (VEGF) production in differentiated THP-1 monocytic cells. Phorbol-12 myristate-13-acetate differentiated THP-1 into macrophage-like cells. 17beta estradiol (10 (-9) M) increased VEGF secretion of controls 3.1-fold in differentiated THP-1 and this effect of 17beta-estradiol was antagonized by dihydrotestosterone, although dihydrotestosterone alone did not alter VEGF secretion. 17beta-estradiol increased steady-state mRNA level of VEGF and the increase was counteracted by dihydrotestosterone in differentiated THP-1, although dihydrotestosterone alone did not alter the VEGF mRNA level. Progesterone did not affect the constitutive and 17beta-estradiol-induced VEGF secretion and mRNA level. Transient transfection revealed that 17beta-estradiol enhanced chloramphenicol acetyl transferase expression driven by VEGF promoter and the enhancement was antagonized by dihydrotestosterone. Adenylate cyclase inhibitor suppressed 17beta-estradiol-induced enhancement of VEGF secretion, mRNA level, and promoter activity, whereas dihydrotestosterone-induced suppression on the effects of 17beta-estradiol was counteracted by 3',5'-adenosine cyclic monophosphate (cAMP) analog. 17beta-estradiol increased intracellular cAMP level by activating adenylate cyclase, while dihydrotestosterone reduced the basal and 17beta-estradiol-increased cAMP level by inhibiting adenylate cyclase. Transfection with 5'-deleted VEGF promoters demonstrated that the region between 88 and -66 bp may be involved in the transcriptional regulation by each hormone. The mutation within activator protein-2 element in this region abrogated the transcriptional stimulation and repression by the respective hormones. 17beta estradiol activated transcription from activator protein-2-responsive reporter plasmid while dihydrotestosterone antagonized the effect of 17beta-estradiol. These results suggest that 17beta-estradiol enhances VEGF production while dihydrotestosterone antagonizes the effect of 17beta-estradiol via up- or downregulation of adenylate cyclase in differentiated THP-1. PMID- 11874494 TI - A novel connexin 30 mutation in Clouston syndrome. AB - Clouston syndrome (hidrotic ectodermal dysplasia) is an autosomal dominant ectodermal dysplasia characterized by alopecia, palmoplantar hyperkeratosis, and nail dystrophy. Recently, mutations in the GJB6 gene encoding the gap junction protein connexin 30 have been shown to cause this disorder. To date, all mutations have involved two codons: G11R and A88V. Here, we report a novel mutation V37E within the first transmembrane domain of connexin 30 in a spontaneous case of Clouston syndrome. The mutation was detected in genomic DNA, confirmed in reverse transcription polymerase chain reaction products, and was excluded from 100 ethnically matched control individuals by restriction enzyme analysis. PMID- 11874495 TI - Evidence for expression of melanocortin-1 receptor in human sebocytes in vitro and in situ. AB - Many lines of evidence indicate that the activity of sebaceous glands can be modulated by neuropeptides. Direct evidence in man, however, is still missing. We show that SZ95 sebocytes, an immortalized human sebaceous gland cell line, express receptors for alpha-melanocyte-stimulating hormone. Reverse transcription polymerase chain reaction with primers against the five melanocortin receptors and immunofluorescence studies using an antibody directed against a peptide corresponding to the amino acids 2-18 of the human melanocortin-1 receptor disclosed specific transcripts and immunoreactivity for melanocortin-1 receptor in these cells. Melanocortin-1 receptor expression was confirmed in sebocytes of normal human skin by immunohistochemistry. In contrast, no immunostaining for the melanocortin-5 receptor could be detected in sebocytes in situ, in accordance with the lack of specific transcripts for this melanocortin receptor in SZ95 sebocytes. As cytokines play an important role in the recruitment of inflammatory cells in acne and related disorders and alpha-melanocyte-stimulating hormone exerts immunomodulatory effects in many other cell types, we investigated the effect of alpha-melanocyte-stimulating hormone on interleukin-8 secretion by SZ95 sebocytes. Treatment with interleukin-1beta resulted in a marked increase in interleukin-8 release that was partially blocked by coincubation with alpha melanocyte-stimulating hormone in a dose-dependent manner. Taken together, we show here that the melanocortin-1 receptor is expressed in vitro and in situ in human sebocytes. By modulating interleukin-8 secretion, alpha-melanocyte stimulating hormone may act as a modulator of inflammatory responses in the pilosebaceous unit. PMID- 11874496 TI - Human skin penetration of flufenamic acid: in vivo/in vitro correlation (deeper skin layers) for skin samples from the same subject. AB - Previously, the interest in in vivo/in vitro correlations in the dermal field of research has increased steadily. Unfortunately, in most cases the skin from different human donors was taken for in vivo and in vitro experiments, which led to problems concerning the interindividual variability of the skin. Therefore, we established a methodology to utilize the same skin for both sets of data. In time dependency, drug amounts in the stratum corneum and the deeper skin layers were determined from eight donors using the same skin area for in vivo and the corresponding in vitro tests. Penetration experiments were carried out with the lipophilic drug flufenamic acid dissolved in wool alcohols ointment as the model formulation, which was administered to the skin under "infinite dose" conditions. At different time points prior to starting the surgery, the drug preparation was applied topically on the edges of the skin area, which was planned for excision using Finn chambers. After anesthetizing the patient and disinfecting the operation area, the incubated skin pieces were cut off first and immediately frozen to limit further drug diffusion. In vitro experiments were performed on the remaining skin flap, using two different test systems, a penetration and a permeation model. At the end of all experiments (in vivo and in vitro) the skin specimens were segmented horizontally and the drug was extracted and quantified. The in vivo and in vitro drug amounts in the stratum corneum and the deeper skin layers, respectively, were compared. The inevitable use of unknown volumes of disinfectant in vivo (medical reasons) might be the reason why a correlation failed for the stratum corneum. Nevertheless, for both in vitro test systems a direct linear correlation was found for the deeper skin layers, which showed slopes of a = 3.2272 +/- 0.3933 (penetration model vs in vivo) and a = 1.7776 +/- 0. 1926 (permeation model vs in vivo). This difference demonstrates the varying influence of the test systems and represents a factor about which in vivo and in vitro data are shifted against each other. As far as the model drug flufenamic acid is concerned, this methodology represents a tool to predict drug penetration into the deeper skin layers in vivo after carrying out corresponding in vitro experiments. Therefore, the potential is given to reduce the number of in vivo experiments, the risk for the volunteers, and the costs for the development of new drug preparations. PMID- 11874497 TI - A novel point mutation in the keratin 17 gene in a Japanese case of pachyonychia congenita type 2. PMID- 11874498 TI - EB simplex superficialis resulting from a mutation in the type VII collagen gene. PMID- 11874499 TI - Analysis of ATP2C1 gene mutation in 10 unrelated Japanese families with Hailey Hailey disease. PMID- 11874501 TI - Increased macrophage migration inhibitory factor (MIF) in the sera of patients with extensive alopecia areata. PMID- 11874500 TI - Unusual pemphigus phenotype in the presence of a Dsg1 and Dsg3 autoantibody profile. PMID- 11874502 TI - Evidence for extensive locus heterogeneity in Naxos disease. PMID- 11874503 TI - The use of histamine in cancer immunotherapy. PMID- 11874505 TI - Bruxism is mainly regulated centrally, not peripherally. AB - Bruxism is a controversial phenomenon. Both its definition and the diagnostic procedure contribute to the fact that the literature about the aetiology of this disorder is difficult to interpret. There is, however, consensus about the multifactorial nature of the aetiology. Besides peripheral (morphological) factors, central (pathophysiological and psychological) factors can be distinguished. In the past, morphological factors, like occlusal discrepancies and the anatomy of the bony structures of the orofacial region, have been considered the main causative factors for bruxism. Nowadays, these factors play only a small role, if any. Recent focus is more on the pathophysiological factors. For example, bruxism has been suggested to be part of a sleep arousal response. In addition, bruxism appears to be modulated by various neurotransmitters in the central nervous system. More specifically, disturbances in the central dopaminergic system have been linked to bruxism. Further, factors like smoking, alcohol, drugs, diseases and trauma may be involved in the bruxism aetiology. Psychological factors like stress and personality are frequently mentioned in relation to bruxism as well. However, research to these factors comes to equivocal results and needs further attention. Taken all evidence together, bruxism appears to be mainly regulated centrally, not peripherally. PMID- 11874506 TI - Use of optical coherence tomography for assessment of dental caries: quantitative procedure. AB - A method for quantitative assessment of dental caries using optical coherence tomography (OCT) was demonstrated. Development of caries lesions in 15 bovine teeth, by demineralization in acidic buffer solution, was quantitatively assessed daily for 3 days, using OCT. An OCT system which can collect A-scans (depth versus reflectivity curve), B-scans (longitudinal images) and C-scans (transverse images at constant depth) was used. While the B- and C-scans qualitatively described the lesion detected, the A-scan which showed the depth (mm) resolved reflectivity (dB) of the tooth tissue was used for the quantitative analysis. After a simple normalization procedure to determine the actual depth the light travelled into the tooth tissue, the area (R) under the A-scan was quantified as a measure of the degree of reflectivity of the tissue. The result showed that R (dB mm) decreased with increasing demineralization time. The percentage reflectivity loss (R%) in demineralized tissue, which related to the amount of mineral loss, was also calculated, and it was observed that R% increased with increasing demineralization time. It was concluded that with the above procedure, OCT could quantitatively monitor the mineral changes in a caries lesion on a longitudinal basis. PMID- 11874507 TI - Effects of diameter, chemical impregnation and hydration on the tensile strength of gingival retraction cords. AB - Gingival retraction cords are useful clinical aids but little information is available regarding desirable physical features that would preserve their integrity under tensile forces. The study aimed to establish under experimental conditions the extent to which tensile strength is affected by variation in cord diameter; impregnation with ferric sulphate (FS) or aluminium sulphate (AS); and cord hydration (wet/dry). Commercial cords and standard cotton cords were assayed in an Instron 1137 machine. Data were analysed using t-test, and one- and four way ANOVA. Results indicated that tensile strength for 200 commercial cords was 1.4950 +/- 1.032 kg; and for 560 cotton cords was 1.2964 +/- 1.4560 kg. Cord hydration had no significant effect on tensile strength, whereas impregnation with AS or FS, a smaller diameter, and/or being a cotton cord decreased tensile strength (P < 0.001). Bivariate analyses showed that hydrated commercial cords had higher tensile strength than dry specimens; hydrated or dry cotton cords were not different. The FS-impregnated cotton cords had lower tensile strength than AS impregnated or control cords, and the effect was greater at higher FS concentrations. This study is one of the first evaluations of the physical properties of cords, highlighting characteristics that may minimize the risk of tearing. PMID- 11874508 TI - Treatment need for fixed metal ceramic bridge prostheses in patients treated by dental students in 1984-1996. AB - Even if implant therapy is very common nowadays, treatment with conventional fixed bridge prosthesis still has indications and cannot be forgotten. Because of improved dental health more teeth can be preserved and more fixed prostheses are prepared also for the elderly. The aim of this study was to discuss the future of treatment need in fixed metal ceramic bridge prostheses based on the analysis of distribution of pontics in dentition in four different age groups during the years 1984-1996. Data were collected from the patient files. The numbers of upper lateral incisors, upper first premolars and lower first molars were analysed in years 1984-1987, 1988-1992 and 1993-1996 between and within age groups of under 34, 35-49, 50-64 years and over 65 years. As a conclusion, in the future the treatment need for fixed bridge prostheses will be highest among patients over 50 years and their most replaced teeth are, besides lower first molars, the upper first premolars. PMID- 11874509 TI - Water sorption and solubility of dental composites and identification of monomers released in an aqueous environment. AB - Water sorption and solubility of six proprietary composite resin materials were assessed, and monomers eluted from the organic matrix during water storage identified. Water sorption and solubility tests were carried out with the following storage times: 4 h, 24 h and 7, 60 and 180 days. After storage, water sorption and solubility were determined. Eluted monomers were analysed by high performance liquid chromatography (HPLC). Correlation between the retention time of the registered peak and the reference peak was observed, and UV-spectra confirmed the identity. The results showed an increase in water sorption until equilibrium for all materials with one exception. The solubility behaviour of the composite resin materials tested revealed variations, with both mass decrease and increase. The resin composition influences the water sorption and solubility behaviour of composite resin materials. The HPLC analysis of eluted components revealed that triethyleneglycol dimethacrylate (TEGDMA) was the main monomer released. Maximal monomer concentration in the eluate was observed after 7 days. During the test period, quantifiable quantities of urethanedimethacrylate (UEDMA) monomer were observed, whereas 2,2-bis[4-(2-hydroxy-3-methacryloyloxypropoxy) phenyl]propane (Bis-GMA) was only found in detectable quantities. No detectable quantities of bisphenol-A were observed during the test period. PMID- 11874510 TI - Defects of dental appearance assessed by patient and dental student groups. AB - A study was carried out to establish the relative need for treatment of a range of defects of dental appearance and identify differences between individual and group assessments. Three groups (n = 42), two of patients (one referred for a defect of dental appearance and one not) and one of dental students, assessed urgency of need on a visual analogue scale (VAS), of four cases of dental defect presented as colour prints. All groups rated the need for treatment of the four cases in the same order. Data sets were non-normal with wide ranges. Patients without a defect of appearance usually assessed treatment need as significantly less urgent. Marked variation was discovered between individual assessments within otherwise similar groups. Based on group decisions, guidelines for treatment of defects of dental appearance could be established. However, so extreme are individual variations that very sympathetic application would be needed if dissatisfaction is not to result. PMID- 11874511 TI - Effect of desensitizers on bond strength of adhesive luting agents to dentin. AB - The current study investigates the influence of three dentin hypersensitivity treating agents (Gluma CPS, MS Coat and Saforide) on bond strength to dentin of two luting agents (Panavia Fluoro Cement and Super-Bond C & B). Sixty bovine dentin substrates were divided into 12 combinations of four treatment conditions (Gluma CPS, MS Coat, Saforide and control) and three adhesive systems (AD Gel sodium hypochlorite + Panavia Fluoro Cement, Panavia Fluoro Cement without AD Gel and Super-Bond C & B). After bonding the treated teeth to steel rods, 24-h tensile bond strengths were determined, and average values (n=5) were compared by analysis of variance (ANOVA). Without application of the desensitizers, bond strengths of the two groups (Super-Bond C & B, 10.2 MPa; AD Gel + Panavia, 11.5 MPa) were comparable, and they were greater than the group bonded with the Panavia material with no AD Gel conditioning (7.1 MPa). Application of the Saforide ammoniated silver fluoride desensitizer reduced bond strength of both the Super-Bond and Panavia luting agents, whereas the MS Coat polymeric agent negatively affected bond strength of the Panavia cement only. The use of the Gluma desensitizer did not affect bond strength of any of the three adhesive systems, and the bond strength of the Panavia cement with the AD Gel conditioning was not reduced by application of any of the three desensitizers. The four combinations of two desensitizers (MS Coat and Gluma CPS) and two adhesive systems (Super-Bond C & B and AD Gel + Panavia) are deemed to be applicable to fixed prosthodontic treatment. PMID- 11874512 TI - Maintenance of condylar position using an occlusal splint after mechanical vibrating-traction of the TMJ. AB - Although adequate relief of excess mechanical loading to the joint has been accepted as one of the important treatment concepts in the orthopaedic field, a treatment method for the temporomandibular joint (TMJ) to relieve excess mechanical loading has not yet been established. This study aimed to clarify the effect of an occlusal splint on the maintenance of the distracted condylar position achieved by vibrating-traction method. Vibrating dynamic traction force was applied for 5 min to the right TMJ using vibrating-traction apparatus. A flat stabilization splint was adjusted to keep the mandibular position and the condylar displacement was evaluated for 6 h after the dynamic traction procedure. Mean vertical displacement of the mandibular right first molar immediately after the vibrating-traction for the six subjects was 156 microm (ranging from 141 to 179 microm). The calculated mean condylar displacement immediately after the traction was 480 mum and could be kept to be 381 mum even after 6 h by wearing the flat stabilization splint. From the results of this study, it was revealed that the mechanically tracted condylar position could be maintained by a flat occlusal splint. It was suggested that the vibrating-traction method followed by the provision of occlusal support might have a possibility to work as a mechanical relieving procedure for the TMJ. PMID- 11874513 TI - Can a prosthesis cause psychological disturbances? AB - This study has the aim of evaluating whether a very simplified, fixed prosthesis and correction (single crown or small bridge) can be the cause of psychological problems in some patients. The tests utilised are, STAI-Y (STAI-S for state anxiety, STAI-T for trait anxiety), BDI for depression, SP-it for stress events, SDS for somatoform disorders, The tests were given to patients at the first examination (t(0)) and at the check-up (t(1)) 1 month after the definitive prosthesis was cemented. The results have shown that somatic preoccupations, in some cases, increased from t(0) to t(1). It is then right that we ask ourselves the following: if the prosthesis were to be very complicated (implants, dentures), what would happen to the patient from a psychological standpoint? PMID- 11874514 TI - The short-term effect of stabilization-type splints on the local asymmetry of masseter muscle sites. AB - Patients with temporomandibular disorders (TMD) may present with a cluster of joint and muscle disorders characterized primarily by pain, joint sounds and irregular or deviating jaw function. Maxillary stabilization-type splints represent the best standard therapy, so the purpose of this study was to evaluate changes in local cross-sectional dimensions (LCSD) of masseter muscle sites associated with short-term application of 'splint therapy' using the diagnostic approach of high-resolution gray-scale ultrasonography. The study included 29 patients who had signs and symptoms of temporomandibular disorders. Ultrasonographic investigation was performed with a linear (B-scan) 7.5 MHz small part transducer to visualize the antero-superior, antero-inferior, medio superior, medio-inferior, postero-superior and postero-inferior sites of the masseter muscle. To assess local muscle asymmetry patterns and to evaluate the respective effect of occluding splints, the 'absolute asymmetry index (AAI)' was used, with the mean maximum muscle diameter of the respective right and left sides calculated from three consecutive measurements before and after splint therapy. Comparing the pre-treatment with the 2-month follow-up values revealed a significant decrease in the overall mean asymmetry indices at the anterior (P < 0.01), medial (P < 0.05) and posterior (P < 0.05) muscle sites. There was no significant change in LCSDs at the various muscle sites (P > 0.05). The results of this study suggest stabilization-type splints to be effective in reducing local muscle asymmetries. Further studies are warranted to evaluate muscle-site specific effects in patient and non-patient groups and to relate these effects to pre-treatment variables like bite force, preferred chewing side, facial morphology and occlusion. PMID- 11874515 TI - Head movement properties during voluntary rapid jaw movement in humans. AB - The purpose of this study was to determine whether the start of the synchronized head movement during mandibular movement is evoked by the peripheral reflexes following mandibular movement (i.e. stretch or trigemino-neck reflexes), or, alternatively, is started by pre-programmed central command. Head movement accompanying voluntary rapid jaw opening movement was studied using accelerometers fixed to the upper and lower incisors, as well as electromyographs (EMGs) of the neck muscles. The direction of head acceleration at the upper incisor was towards head extension at the beginning of jaw opening movement in 89.2% of all trials, opposite to the direction of lower jaw acceleration. The onset of head acceleration was later than that of the lower jaw acceleration by averages of 6.2-10.7 ms, and the onset of electromyographic activities of the sternocleidomastoid (SCM) muscle preceded that of head acceleration by an average of 12.5-24.3 ms. These findings suggest that head movement during mandibular movement is not started by peripheral reflexes but by pre-programmed central commands. This may be relevant to muscular discomfort in the neck and shoulder regions of patients with stomatognathic disorders. PMID- 11874516 TI - Bonding strength between a hard chairside reline resin and a denture base material as influenced by surface treatment. AB - Direct relining of dentures made with hard chairside reline resins is faster than laboratory-processed reline systems and the patient is not without the prosthesis for the time necessary to perform the laboratory procedures. However, a weak bond between the autopolymerizing acrylic reline resins and the denture base material has been observed. This study evaluated the effect of six different surface treatments on the bond strength between a hard chairside reline acrylic resin and a heat-cured acrylic resin. Specimens of the heat-cured acrylic resin were divided into seven groups. One of these groups remained intact. In the other groups, a 10-mm square section was removed from the centre of each specimen. The bonding surfaces were then treated with (i) methyl methacrylate monomer, (ii) isobutyl methacrylate monomer, (iii) chloroform, (iv) acetone, (v) experimental adhesive and (vi) no surface treatment -- control group. Kooliner acrylic resin was packed into the square sections and polymerized. The bonding strength was evaluated by a three-point loading test. The results were submitted to one-way analysis of variance (ANOVA) followed by a Tukey multiple range test at a 5% level of significance. No significant difference was found between the surface treatment with Lucitone 550 monomer or chloroform, but both were stronger than the majority of the other groups. The bond strength provided by all the surface treatments was lower than that of the intact heat-cured resin. PMID- 11874517 TI - Effectiveness of exercise therapy in patients with internal derangement of the temporomandibular joint. AB - This study intended in evaluating the effectiveness of exercise therapy in patients with craniomandibular disorders (CMD). Twenty consecutive patients suffering from CMD with anterior disc displacement without reduction consulting a CMD service were included in the study if they met following criteria: (i) pain in the temporomandibular region, (ii) reduced incisal edge clearance (<35 mm), (iii) magnet resonance imaging confirmed anterior disc displacement without reduction and (iv) evidence of postural dysfunction. All patients were assigned to a waiting list, serving as a no-treatment control period, according to a before-after trial. The treatment consisted of active and passive jaw movement exercises, correction of body posture and relaxation techniques. A total of 18 patients completed the study, no adverse effects occurred. Following main outcome measures were evaluated: (1) pain at rest (2) pain at stress (3) impairment (4) mouth opening at base-line, before and after treatment and at 6 month follow-up. As a result of treatment pain, impairment and mouth opening improved significantly more than during control period (paired samples t-test P < 0.05). After treatment four patients had no pain at all (chi-square: P < 0.05) and only seven patients revealed an impaired incisal edge clearance after treatment. (chi square Test, P < 0.001). At follow up, seven patients had no pain and experienced no impairment. Exercise therapy seems to be useful in the treatment of anterior disc displacement without reduction. PMID- 11874518 TI - Electromyographic analysis of fatigue in temporalis and masseter muscles during continuous chewing. AB - The aim of this study was to evaluate the electromyographic changes in the anterior temporalis and masseter muscles after fatigue induced by continuous chewing. Surface electrodes were placed bilaterally over the anterior temporalis and masseter muscles of 31 subjects with normal dentition. Recordings were taken every minute from the beginning of chewing to the exact moment the volunteer reported subjective sensation of fatigue in the jaw muscles. The length of endurance period (fatigue threshold) was measured to each subject, as well as the average amplitude and duration of activation phase and duration of relaxation phase through electromyographic recording of each muscle. The average endurance period was about 500 s. No significant change occurred on the average amplitude of activation phase meanwhile the duration of both the activation and relaxation phase diminished after fatigue. PMID- 11874519 TI - The efficiency of dentine adhesives in treating non-caries cervical lesions. AB - The aim of this study was to prove the hypothesis that dentine adhesives can be used as therapeutic material in treating dentine hypersensitivity, regardless of aetiology. The research was conducted on 492 students of Zagreb University School of Dental Medicine (154 male, 338 female), 20-25-years-old. The defects of the tooth neck were found in 38 patients, on 133 teeth (38 of male patients, 95 of female) and dentine hypersensitivity was recorded according to the subjective sensations of patients. Therapeutic characteristics of three adhesive materials were examined simultaneously: All Bond 2, fourth generation adhesive, Syntac Single Component and One Step, fifth generation adhesives. Teeth treated with dentine lacquer Cervitec were used as a control group. Tooth necks were treated with selected materials according to instructions of a manufacturer. A completed statistical survey of the results has clearly shown that dentine adhesives can be used in symptomatic therapy of dentine hypersensitivity of the non-caries cervical lesions (NCCL). The survey has also shown that dentine adhesives of the fifth generation (Syntac Single Component and One Step) have much higher efficiency rate than dentine adhesives of the fourth generation (All Bond 2) and dentine lacquer (Cervitec). According to data complied through this study the conclusion can be brought forward that dentine adhesives are not a final solution to the problem of dentine hypersensitivity, because their efficiency decreases with time. PMID- 11874520 TI - Comparative serum pharmacokinetics of the fluoroquinolones enrofloxacin, difloxacin, marbofloxacin, and orbifloxacin in dogs after single oral administration. AB - The pharmacokinetics after oral application of the fluoroquinolones (FQs), enrofloxacin, difloxacin, marbofloxacin and orbifloxacin were compared in independent crossover studies in Beagle dogs. Commercially available tablet formulations were given at common dosage recommended by the manufacturers which were 2.0 mg/kg body weight (bw) for marbofloxacin, 2.5 mg/kg bw for orbifloxacin and 5.0 mg/kg bw for enrofloxacin and difloxacin. Analysis was performed by an agar diffusion assay. Pharmacokinetic parameters were calculated by noncompartmental methods. All FQs were rapidly absorbed and achieved average peak serum concentrations of 1.41, 1.11, 1.47 and 1.37 mug/mL for enrofloxacin, difloxacin, marbofloxacin and orbifloxacin, respectively. Enrofloxacin was eliminated at a terminal half-life (t(1/2)) of 4.1 h, difloxacin at 6.9 h, orbifloxacin at 7.1 h and marbofloxacin at 9.1 h. While the area under the serum concentration-time curve of the 24-h dosing interval (AUC0--24) for marbofloxacin and orbifloxacin were similar (approximately 13 microg x h/mL), enrofloxacin attained an AUC(0-24) of 8.7 and difloxacin of 9.3 microg x h/mL. Because of its favourable pharmacokinetics combined with excellent in vitro activity, enrofloxacin exhibited superior pharmacodynamic predictors of in vivo antimicrobial activity as C(max)/MIC (maximum serum concentration/minimum inhibitory concentration) and AUC(0-24)/MIC (area under the 24-h serum concentration--time curve/minimum inhibitory concentration) compared with other FQs. PMID- 11874521 TI - Plasma achiral and chiral pharmacokinetic behaviour of intravenous oxfendazole co administered with piperonyl butoxide in sheep. AB - Co-administration of piperonyl butoxide (PB) potentiates fenbendazole (FBZ) in small ruminants. The resultant increase in bioavailability of FBZ and its metabolite oxfendazole (OFZ) has important implications for the efficacy of these drugs against benzimidazole (BZD)-resistant strains of Teladorsagia circumcincta. This study evaluated the racemic (achiral) and enantiomeric (chiral) plasma disposition kinetics of OFZ and its metabolites after the co-administration of PB and OFZ in sheep. Six 6-8-month-old, parasite-free, female Dorset sheep (30-40 kg) were used in a two-phase crossover experiment. In phase I, three sheep received 30 mg/kg PB orally, followed by a single intravenous (i.v.) injection of OFZ at 5 mg/kg. The other three animals were treated similarly except that 5 mL of water replaced PB. In phase 2, treatments for the two groups were reversed and were given 14 days after the initiation of phase I. Three analytes OFZ, FBZ and fenbendazole sulphone (FBZSO(2)) were recovered in plasma up to 48 h post treatment in both experimental groups. Achiral and chiral pharmacokinetic (PK) profiles for OFZ, after the co-administration of PB, were characterized by a significantly greater area under the concentration--time curve (AUC) and a longer mean residence time (MRT). Chiral OFZ distribution ratios were comparable in both treatment groups. Piperonyl butoxide treatment markedly influenced the plasma PK profiles for FBZ and FBZSO(2) following OFZ administration. Production of FBZ was enhanced as reflected by increased (> 60%) AUC, delayed T(max) and a significantly delayed (> 45%) elimination (t(1/2)(el)). Although AUC values for FBZSO(2) were not significantly different between groups, this metabolite was depleted more slowly from plasma (t(1/2)(el) > 60% and MRT > 42%) following PB treatment. This study demonstrated that PB co-administration is associated with an inhibition of OFZ biotransformation, as evidenced by the significantly higher plasma concentrations of OFZ and FBZ, and this could have important implications in terms of anti-parasite therapy against BZD-resistant parasite strains. PMID- 11874522 TI - In vitro ruminal biotransformation of benzimidazole sulphoxide anthelmintics: enantioselective sulphoreduction in sheep and cattle. AB - The comparative in vitro sulphoreduction of the (+) and (-) enantiomers of albendazole sulphoxide (ABZSO) and oxfendazole (OFZ) by ruminal fluid obtained from sheep and cattle, was investigated, under anaerobic conditions, in this study. Ruminal fluid samples were obtained from Holstein steers fitted with a permanent rumen fistula and from Corriedale lambs via an oesophageal tube. Albendazole sulphoxide, incubated as either the racemic (rac) mixture or as each individual enantiomeric form, was extensively sulphoreduced to form albendazole (ABZ) by ruminal fluid from both species. The concentrations of ABZ formed at different incubation times were between 55 and 158% greater after the incubation of cattle ruminal fluid with (+) ABZSO, compared with that produced when (-) ABZSO was the incubated substrate. Similarly, the concentrations of ABZ were 1.3- 3.0-fold higher when (+) ABZSO was incubated with sheep ruminal fluid. Significantly higher rates of depletion were observed for the (+) enantiomeric form when ABZSO was incubated with ruminal fluid from both species. The rates of ABZ formation from both ABZSO enantiomeric forms were significantly higher in sheep compared with cattle ruminal fluid. Fenbendazole (FBZ) was the metabolite formed after the incubation of the racemic form of OFZ with ruminal fluid obtained from both species. The metabolic profile of both OFZ enantiomers followed a similar pattern to that observed for ABZSO enantiomers. A bi directional chiral inversion of one enantiomer into its antipode was observed. The (+) enantiomer appeared in the incubation medium when (-) ABZSO was the incubated substrate, and also the (-) antipode was detected after (+) ABZSO incubation with ruminal fluid obtained from both species. The results reported here demonstrate an enantioselective ruminal sulphoreduction of ABZSO and OFZ (substrate enantioselectivity). These findings contribute to interpret the chiral behaviour of benzimidazole-sulphoxide anthelmintics. PMID- 11874523 TI - Serum pharmacokinetics and tissue and milk residues of oxytetracycline in goats following a single intramuscular injection of a long-acting preparation and milk residues following a single subcutaneous injection. AB - Separate groups of goats were used to determine drug depletion patterns in serum (n=10), tissue (n=20) and milk (n=8) following a single intramuscular (i.m.) dose of 20 mg/kg of a long-acting oxytetracycline (OTC) formulation (Liquamycin LA 200). Milk residues were also determined following a subcutaneous (s.c.) administration of the same product at the same dose. Serum samples were taken for 24 h post-treatment and tissues (fat, liver, kidney, muscle and injection site) collected at 4, 7, 14, 21 and 28 days following injection. Milk from lactating goats was collected every 12 h for 8 days following both the i.m. and s.c. treatments utilizing an intervening 5-week washout period. Residues in serum and tissue were measured using a microbial inhibition assay, while milk residues were measured using both a microbial inhibition assay and a validated HPLC method. The serum pharmacokinetic parameters of OTC in goats were determined, with a mean AUC=67.4 microg h/mL, mean terminal half-life=14.4 h, and apparent clearance=0.33 L/kg h. Tissue half-lives could not be determined with confidence because the collection times provided only two points at which residues could be measured for most tissues. Oxytetracycline residues in all goat tissue samples measured less then cattle tissue tolerance by 96 h postdosing. One-compartment model describing milk depletion data for i.m. and s.c. dosing had terminal slope half-lives of 20.1 and 36.1 h, respectively. By 96 h post-treatment none of the milk samples contained OTC residues in excess of the cattle milk tolerance (0.3 p.p.m.). For both milk and tissue, the upper-bound 99% confidence intervals for the samples taken from goats 96 h postdosing were lower than approved cow milk and tissue tolerances. PMID- 11874524 TI - Pharmacokinetics of ceftiofur in plasma and uterine secretions and tissues after subcutaneous postpartum administration in lactating dairy cows. AB - A study was conducted to measure concentrations of potentially active ceftiofur derivatives, in plasma, in uterine tissues (endometrium and caruncles) and in uterine secretions at different time points after a single subcutaneous administration of ceftiofur hydrochloride (Excenel RTU Sterile Suspension) at the dose of 1 mg/kg body weight in Holstein-Friesian dairy cows. The animals (n=4) were injected within 24 h of calving, after expulsion of the foetal membranes. Plasma, lochial fluid, caruncles and endometrium were collected before ceftiofur hydrochloride administration and at 1, 2, 4, 8, 12 and 24 h after treatment. For each cow the concentrations of ceftiofur in the biological matrices were quantified using an high-performance liquid chromatography (HPLC) assay. The limit of quantification of the method was 0.1 microg/mL for plasma and 0.1 microg/g for lochial fluid, caruncles and endometrium. The concentrations of potentially active ceftiofur derivatives detected in plasma reached a maximum of 2.85 +/- 1.11 microg/mL at 2 h and decreased to 0.64 +/- 0.14 microg/mL at 24 h after administration. In lochial fluid, these concentrations reached a maximum of 0.97 +/- 0.25 microg/g at 4 h and decreased to 0.22 +/- 0.21 microg/g at 24 h after administration. In endometrium, these concentrations reached a maximum of 2.23 +/- 0.82 microg/g at 4 h and decreased to 0.56 +/- 0.14 microg/g at 24 h following the injection, whereas these levels in caruncles were 0.96 +/- 0.45 and 0.60 +/- 0.39 microg/g obtained at 8 and 24 h, respectively. At the dose of 1 mg/kg body weight in healthy dairy cows, subcutaneous administration of ceftiofur (as ceftiofur hydrochloride) after parturition results in concentrations of ceftiofur derivatives in uterine tissues and in lochial fluid that exceed the reported minimal inhibitory concentrations (MICs) for the common pathogens (Escherichia coli, Fusobacterium necrophorum, Bacteroides spp., and Arcanobacterium pyogenes) associated with acute puerperal metritis. PMID- 11874525 TI - Some factors influencing the level of clinical sedation induced by medetomidine in rabbits. AB - Rabbits (n=23) received intravenous bolus medetomidine at 100 mug/kg. Prior to medetomidine administration, heart and respiratory rates were measured, arterial blood was collected and analysed for plasma cortisol, glucose and albumin concentrations. Fifteen minutes after medetomidine administration, heart and respiratory rates were measured again and sedation was scored. The rabbit was afterwards anaesthetized with 20 mg/kg ketamine administered intravenously to enable spinal tap and heart puncture. Cerebrospinal fluid (CSF) was collected (this occurred 20 min post medetomidine administration) and analysed for medetomidine concentration. Blood was collected by heart puncture immediately after the spinal tap and analysed for serum medetomidine concentration. Cerebrospinal fluid medetomidine concentration correlated negatively with sedation. Serum medetomidine correlated positively with CSF medetomidine concentration. Cerebro-spinal fluid medetomidine was 17 +/- 13% of serum medetomidine concentration. Plasma cortisol and glucose concentrations correlated negatively with serum medetomidine. We conclude that after an intravenous bolus administration of a low sedative dose of medetomidine to rabbits; CSF concentration of the drug correlate negatively with sedation and that this may be because of the fact that only the free and unbound medetomidine may be available for detection in the CSF, the concentration of medetomidine detected in the CSF was much lower than that in blood and a positive correlation exists between CSF and serum medetomidine concentrations. Stress may have some effect on the distribution or metabolism of medetomidine in rabbits. PMID- 11874526 TI - The pharmacokinetics, metabolism and urinary detection time of etamiphylline in camels after intramuscular administration. AB - The pharmacokinetics of etamiphylline were determined after an intramuscular (i.m.) dose of 3.5 mg/kg body weight in six healthy camels. Furthermore, the metabolites and drug detection time were evaluated. The data obtained median and (range) were as follows: the terminal elimination half-life (t(1/2 beta), h) was 3.04 (2.03-3.62); apparent total body clearance (Cl/F, L/h/kg) was 1.27 (0.74 2.99); the apparent volume of distribution at steady state (V(ss)/F, L/kg) was 4.94 (3.57-12.54); and renal clearance (Cl(r), L/h/kg) determined in two camels was 0.005 and 0.004, respectively. The detection time of etamiphylline in urine after an i.m. dose of 3.5 mg/kg body weight ranged between 12 and 13 days. Three etamiphylline metabolites were tentatively identified in camels urine: The first one desethyletamiphylline was the main metabolite and resulted from N deethylation of etamiphylline had a molecular weight of 251, and was detected in urine for about 13-14 days. Theophylline (molecular weight 180) was the second metabolite and resulted from ring N-dealkylation of etamiphylline. It was present in small amounts and was detected for about 5 h after drug administration in urine. The third metabolite, possibly resulted from demethylation of etamiphylline, had a molecular weight of m/z 265, and was present in small amounts and was detected in urine for about 5 h after drug administration. PMID- 11874528 TI - Residue depletion of thiamphenicol in the sea-bass. AB - The residue depletion of thiamphenicol (TAP) was investigated in the sea-bass (Dicentrarchus labrax) after 5 days' treatment with medicated food at a dose of 15 or 30 mg/kg bw/day. Fish were sampled for blood and muscle + skin from 3 h until 14 days after treatment. Thiamphenicol concentrations were assayed by high performance liquid chromatography. Thiamphenicol concentrations measured 3 h after stopping treatment were 0.77 microg/mL and 0.91 (15 mg/kg dose) or 1.32 microg/mL and 1.47 microg/g (30 mg/kg dose), in plasma and muscle + skin, respectively. After a withdrawal of 3 days, plasma and tissue concentrations were: 0.08 microg/mL and 0.03 microg/g (lower dose) or 0.12 microg/mL and 0.06 microg/g (higher dose), respectively. Thiamphenicol was not detectable either in plasma or in tissues on days 7, 10 and 14 following withdrawal of the medicated food. Based on maximum residue levels (MRL) for TAP in fin fish, established at 50 microg/kg for muscle and skin in natural proportions, a withdrawal period of 5 and 6 days is proposed, after treatment at 15 or 30 mg/kg of TAP with medicated feed pellets, respectively, to avoid the presence of violative residues in the edible tissues of the sea-bass. PMID- 11874527 TI - 5-Hydroxytryptamine mediated contractions in isolated preparations of equine ileum and pelvic flexure: pharmacological characterization of a new 5-HT(4) agonist. AB - The effects of 5-hydroxytryptamine (5-HT), HTF 919, a new 5-HT(4) agonist, and the antagonists SB 203-186 (5-HT(4)) and tropisetron (5-HT(3)) on intestinal motility were tested in vitro on isolated preparations of horse ileum and pelvic flexure. Concentration-response curves were created by cumulative application of the agonists with or without preincubation of the antagonists. The 5-HT preparation induced a concentration-dependent contraction in equine ileum and pelvic flexure. The results indicate that 5-HT receptors are present in all parts of equine intestine investigated in this study. Tropisetron was found to act as a noncompetitive antagonist in all locations of the equine intestine. SB 203-106 was confirmed as an antagonist to 5-HT in the equine ileum circular muscle, in pelvic flexure circular and longitudinal muscle. Nevertheless, a discernible increase of smooth muscle contractions caused by HTF 919 could only be observed in pelvic flexure. In accordance with an earlier study in the guinea pig, in the equine gut HTF 919 acted as a partial agonist for the 5-HT(4) receptor with an affinity constant in the nanomolar range. It is concluded that 5-HT receptors, and especially their subtypes, may represent a promising target for the treatment and prevention of gastrointestinal (GI) motility disorders in horses. PMID- 11874529 TI - Effect of acepromazine and butorphanol on isoflurane minimum alveolar concentration in goats. PMID- 11874530 TI - Modulation by levamisole of CD45RA and CD45RC isoforms expression in the gut of weaned pigs vaccinated against colibacillosis. PMID- 11874531 TI - Pharmacokinetics of ceftriaxone administered by the intravenous, intramuscular or subcutaneous routes to dogs. AB - The purpose of this study was to investigate the pharmacokinetics of ceftriaxone after single intravenous (i.v.), intramuscular (i.m.) and subcutaneous (s.c.) doses in healthy dogs. Six mongrel dogs received ceftriaxone (50 mg/kg) by each route in a three-way crossover design. Blood samples were collected in predetermined times after drug administration. Results are reported as mean +/- standard deviation (SD). Total body clearance (Cl(t)) and apparent volume of distribution (V(z)) for the i.v. route were 3.61 +/- 0.78 and 0.217 +/- 0.03 mL/kg, respectively. Terminal half-life harmonic mean (t(1/2 lambda)) was 0.88; 1.17 and 01.73 h for the i.v., i.m and s.c. routes, respectively. Mean peak serum concentration (C(max)) was 115.10 +/- 16.96 and 69.28 +/- 14.55 microg/mL for the i.m and s.c. routes, respectively. Time to reach C(max) (t(max)) was 0.54 +/- 0.24 and 1.29 +/- 00.64 h for the i.m and s.c. routes, respectively. Mean absorption time (MAT) was 1.02 +/- 0.64 and 2.23 +/- 00.73 h for the i.m and s.c. routes, respectively. Bioavailability was 102 +/- 27 and 106 +/- 14% for the i.m and s.c. routes, respectively. Statistically significant differences were determined in C(max), t(max), MAT and t(1/2 lambda) of s.c. administered ceftriaxone when compared with the i.v and i.m. routes. These findings suggest that once or twice s.c. or i.m. daily administered ceftriaxone should be adequate to treat most susceptible infections in dogs. PMID- 11874535 TI - Development of a high-volume aerosol collection system for the identification of air-borne micro-organisms. AB - AIMS: A high-volume aerosol collector was developed to efficiently capture airborne bacteria in order to assess levels of diversity in the air. METHODS AND RESULTS: Particulate matter was collected on a device designed to filter 1.4 x 10(6) litres of air in a 24 h period on a 1-microm pore size polyester membrane. Methods were optimized for extraction of genomic DNA from the air filter concentrate. Preparation times of 90 s with 0.5-0. 05 mm diameter zirconia/silica beads yielded the highest concentration genomic DNA that was able to support PCR. A 24-h air sample was taken in Salt Lake City, Utah and the microbial composition was determined by the amplification and sequence analysis of 16S ribosomal DNA fragments. CONCLUSIONS: Sequence analysis revealed a large diversity in the type of microbial species present including clones matching the sequence of Clostridium botulinum. The primary components of the aerosol sample included many different spore-forming bacteria as well as more fragile members of the Proteobacteria division. SIGNIFICANCE AND IMPACT OF STUDY: The high-volume air collection and genomic DNA recovery system allows for the rapid detection of both cultivable as well as culture-resistant organisms in the environment. PMID- 11874534 TI - Molecular analysis of Vibrio parahaemolyticus isolated from human patients and shellfish during US Pacific north-west outbreaks. AB - AIMS: The objective of this study was to investigate the occurrence and distribution of haemolysin genes, plasmid profile, serogroup analysis and cellular urease activity for Vibrio parahaemolyticus isolates from infected human patients and oysters from the Pacific north-western United States between 1988 and 1997. METHODS AND RESULTS: All of the clinical and environmental isolates tested in this study exhibited the presence of the thermolabile haemolysin gene, tl, confirming that all of the isolates were V. parahaemolyticus. Furthermore, the V. parahaemolyticus isolates that contained either the thermostable direct haemolysin gene, tdh, or the thermostable direct haemolysin-related gene, trh, or both, were also positive for urease. Isolates from infected human patients belong to serogroups O1 and O4, whereas, the isolates from oysters belong to serogroups O1, O4 and O5. These results suggest that the presence of a V. parahaemolyticus serogroup O1 and O4 could indicate the presence of a virulent strain of this pathogen. In this study, the presence of the haemolysin genes, serogroup profiles and urease production in V. parahaemolyticus isolated from human patients correlated with the oysters collected during the outbreaks. However, no significant correlation of the plasmid profiles was detected, based on their distribution and molecular weights, between V. parahaemolyticus isolated from infected human patients and from oysters collected during this outbreak. CONCLUSIONS, SIGNIFICANCE AND IMPACT OF STUDY: It is apparent from this study that the identification of the haemolysin genes by multiplex PCR amplification, in conjunction with serogroup analysis and urease production, can be used to monitor shellfish for the presence of potentially pathogenic strains of V. parahaemolyticus. PMID- 11874536 TI - Chitosan potentiates the antimicrobial action of sodium benzoate on spoilage yeasts. AB - AIMS: The objective of this study was to determine whether low concentrations of chitosan and benzoate in combination could be used to enhance the antimicrobial action of either compound alone against three spoilage yeasts in saline solutions. METHODS AND RESULTS: Saccharomyces exiguus, Saccharomycodes ludwigii and Torulaspora delbrueckii were suspended in 0.05 and 0.005% chitosan glutamate and 0.025% sodium benzoate, alone or in combination, in 0.9% saline solutions at pH 6.2 and 4.5. Survivor curves were constructed from viable counts determined periodically for up to 120 min. Chitosan at 0.005% almost doubled the extent of death caused by 0.025% benzoate alone, from about 1-2 log to about 2-4 log cfu ml(-1), depending on pH and target organism. CONCLUSIONS: Chitosan (0.005%) and 0.025% sodium benzoate acted synergistically against spoilage yeasts in saline solutions. SIGNIFICANCE AND IMPACT OF THE STUDY: These results suggest that the natural compound chitosan may be useful as an adjunct in the potentiation of the biocidal efficacy of antimicrobial compounds such as benzoates. PMID- 11874538 TI - Production of an exopolysaccharide bioflocculant by Sorangium cellulosum. AB - AIMS: To isolate a new exopolysaccharide bioflocculant produced by the myxobacterium Sorangium cellulosum NUST06, and to characterize its chemical composition and expolysaccharide production relative to carbon source. METHODS AND RESULTS: Exopolysaccharide levels and biomass production by S. cellulosum NUST06 were analysed relative to carbon source. Glucose in the medium at a level of 3 g l(-1) completely inhibited cell growth and exopolysaccharide production, but low concentrations of glucose (1-2 g l(-1)) could stimulate cell utilization of starch. The chemical composition and flocculating activity of the NUST06 exopolysaccharide was investigated. The flocculant comprised 38.3% proteins and 58.5% carbohydrates, of which glucose, mannose and glucuronic acid were present at 51.3%, 39.2% and 10.5%, respectively. The flocculating activity of the NUST06 flocculant depended strongly on cations. CONCLUSIONS: It is feasible to produce an exopolysaccharide bioflocculant by the strain NUST06 in a mineral salts medium using starch as a carbon source. SIGNIFICANCE AND IMPACT OF THE STUDY: This strain may be advantageous for commercial bioflocculant production and may enrich existing knowledge of myxobacteria. PMID- 11874537 TI - Microbial colonization of naturally black olives during fermentation and associated biochemical activities in the cover brine. AB - AIMS: To establish the site of microbial growth on naturally black fermented table olives, and to monitor the population dynamics of yeasts and selected micro organisms together with the changes in organic acid profile and pH in the cover brine during fermentation. METHODS AND RESULTS: During fermentation, the numbers of Enterobacteriaceae and Pseudomonas spp. in the brine decreased whilst lactic acid bacteria and yeast populations increased. Scanning electron microscopy showed that a yeast-rich biofilm developed on the epicuticular wax of the olive skin during fermentation. Yeasts also predominated in the stomatal openings, but bacteria were more numerous in intercellular spaces in the sub-stomatal flesh. Citric, malic and tartaric acids were the major organic acids accumulating in the brine during fermentation. CONCLUSIONS: Micro-organisms associated with the skin, stomata and flesh in fermenting black olives may experience different local conditions to those prevailing in the cover brine. SIGNIFICANCE AND IMPACT OF THE STUDY: These are the first observations of the micro-organisms associated with the fruit of naturally fermented black olives and of the accumulation of specific organic acids during fermentation. PMID- 11874539 TI - Involvement of RNA and DNA in the staining of Escherichia coli by SYTO 13. AB - AIMS: To assess the extent to which DNA and RNA bacterial content contributes to fluorescent response of SYTO 13. METHODS AND RESULTS: RNA and DNA of Escherichia coli 536 cells were extracted and fluorimetrically quantified to compare the different contents, throughout a 24 h culture, with their SYTO 13 fluorescence emission when analysed by the cytometer. SYTO 13 fluorescence varied depending on the stage of bacterial growth and in accordance with both DNA and RNA content. RNA content accounted for at least two-thirds of the total fluorescence of a cell. Escherichia coli cells were treated with chloramphenicol to improve their RNA content. With this treatment, both nucleic acids remained constant but there was a clear improvement in fluorescent emission. SYTO 13 fluorescence was also studied in E. coli X-1488 minicells. CONCLUSIONS: Although both nucleic acids are implicated, RNA accounts for a major part of SYTO 13 fluorescence. The fluorescence cannot be considered as a direct reflection of nucleic acid content. Other factors, such as topology or supercoiling, need to be considered. SIGNIFICANCE AND IMPACT OF THE STUDY: The results confirm the efficacy of SYTO 13 for labelling bacteria and for assessing the distinct physiological status. A better knowledge of the parameters implicated in its fluorescence emission has been achieved. PMID- 11874540 TI - Detection and potential indicators of the presence of hepatitis C virus on surfaces in hospital settings. AB - AIMS: The risk of hepatitis C virus infection in hospital environments can be assessed not only by studying epidemiological data and work practices, but also by the detection of these viruses (or indicators thereof) in health-care settings, on instruments etc. METHODS: Since standardized techniques specific to this end do not exist, this study was undertaken to apply methods currently used on clinical samples to the assessment of environmental HCV risk, either through direct detection of the virus (RT-PCR), or by probing for haemoglobin as a potential indicator of blood contamination. The tested techniques were applied in a trial environmental monitoring programme undertaken in various hospital laboratories and clinics, during which total bacterial count determinations were performed in parallel with haemoglobin and hepatitis C virus detection. SIGNIFICANCE AND IMPACT OF THE STUDY: The data indicate that the applied methods are of value in detecting low levels of contamination in a hospital environment. PMID- 11874541 TI - The occurrence and properties of Bacillus thuringiensis isolated from free-living animals. AB - AIMS: To assess the prevalence and properties of Bacillus thuringiensis isolated from the intestines of small mammals. METHODS AND RESULTS: Bacillus thuringiensis was found in 11% of rodents and 17% of insectivores. Using PFGE of chromosomal DNA, SDS-PAGE of whole-cell proteins and biochemical tests (API system), 12 isolates and three reference strains were classified. Numerical analysis revealed 61% and 89% similarity of protein profiles and biochemical properties of the bacilli, respectively. The results of PFGE were consistent with the outcomes of the analysis of protein profiles. CONCLUSIONS: Although B. thuringiensis is not common in the intestines of small mammals, it is heterogeneous at the genotypic and phenotypic level. SIGNIFICANCE AND IMPACT OF THE STUDY: The results presented here help to explain the diversity and the ecological significance of B. thuringiensis. Future study should focus on the toxic activity of the isolated strains. PMID- 11874542 TI - Immunoassay method for quantitative determination of nisin in solution and on polymeric films. AB - AIMS: To develop a non-competitive direct immunoassay method for quantifying nisin in solution or adsorbed to surfaces. METHODS AND RESULTS: The developed method differs from traditional ELISA in pre-forming a complex of the nisin antibodies and the secondary antibody conjugate before addition to the nisin coated solid phase. The modifications may help decrease interference from the nisin degradation molecules and reduce non-specific adsorption of secondary antibody conjugate. The new method has a detection limit of 65 ng nisin and correlates well (r=0.982) with the agar diffusion bioassay. CONCLUSIONS: A new sensitivity method was developed to determine the amounts of nisin adsorbed to a polymeric surface. SIGNIFICANCE AND IMPACT OF THE STUDY: The new technique can provide a reproducible and accurate method to quantitate nisin molecules in solutions and those bound on polymeric surfaces, and it is free of the limitations of the traditional agar diffusion bioassay. PMID- 11874543 TI - Immobilization of fructosyltransferase from Streptococcus mutans on hydroxyapatite surfaces induces the formation of multimeric complexes. AB - AIMS: To investigate the formation of fructosyltransferase (FTF) complexes on hydroxyapatite (HA) surfaces. METHODS AND RESULTS: Cell-free extracellular FTF from Streptococcus mutans, purified from hyperproducing strain V-1995, was adsorbed onto HA and then eluted from the surface by means of a concentration gradient of potassium phosphate buffer. The FTF monomers loaded onto HA formed, upon adsorption, various complexes ranging from 200 to 700 kDa as demonstrated using native PAGE. All these complexes exhibited enzymatic activity. CONCLUSIONS: Adsorption of FTF onto HA induced the formation of stable and enzymatically active complexes. SIGNIFICANCE AND IMPACT OF THE STUDY: The formation of these complexes may explain the change of FTF catalytic properties after adsorption onto HA. This study is another step in determining the properties of a-cellular constituents of the oral biofilm. PMID- 11874544 TI - A rapid and efficient assay for extracting DNA from fungi. AB - AIMS: A method for the rapid extraction of fungal DNA from small quantities of tissue in a batch-processing format was investigated. METHODS AND RESULTS: Tissue (< 3.0 mg) was scraped from freshly-grown fungal isolates. The tissue was suspended in buffer AP1 and subjected to seven rounds of freeze/thaw using a crushed dry ice/ethanol bath and a boiling water bath. After a 30 min boiling step, the tissue was quickly ground against the wall of the microfuge tube using a sterile pipette tip. The Qiagen DNeasy Plant Tissue Kit protocol was then used to purify the DNA for PCR/sequencing applications. CONCLUSIONS: The method allowed batch DNA extraction from multiple fungal isolates using a simple yet rapid and reliable assay. SIGNIFICANCE AND IMPACT OF THE STUDY: Use of this assay will allow researchers to obtain DNA from fungi quickly for use in molecular assays that previously required specialized instrumentation, was time-consuming or was not conducive to batch processing. PMID- 11874545 TI - Antibiotic susceptibility of bacterial strains isolated from patients with various infections. AB - AIMS: Isolates from various samples obtained during 1998 and 1999 were identified and their susceptibility to third-generation cephalosporins, monobactams and/or cephamycins studied along with any production of ESBLs. METHODS AND RESULTS: Of these samples, bacteria most frequently isolated by the conventional techniques and Vitek GNI card were Escherichia coli (37%), Klebsiella pneumoniae (27%) and Enterobacter cloacae (16%). Using disk diffusion and double-disk synergy tests, we found that 71% strains produced ESBLs and 18% strains produced ESBLs and cephamycinases. Banding patterns of PCR amplification with the designed primers showed that 57% strains were capable of harbouring bla(SHV) genes. The bla(TEM), bla(CMY) and bla(AmpC) genes were harboured by 55%, 31% and 12% strains, respectively. Forty-five percent of strains contained more than two types of beta lactamase genes. In particular, one strain contained bla(TEM), bla(SHV), bla(CMY) and bla(AmpC) genes. CONCLUSIONS: The percentage of ESBL-producing strains was high. The most prevalent beta-lactamase gene was bla(SHV) gene. The bla(CMY) genes have been prevalent in cephamycin-resistant strains. The multidrug resistant strains resistant to third-generation cephalosporins and cephamycins were detected in high percentage. SIGNIFICANCE AND IMPACT OF THE STUDY: Resistance mechanisms to beta-lactams, comprising mostly extended-spectrum beta lactamase (ESBL) production, lead to the resistance against even recently developed beta-lactams in enterobacteria, which is now a serious threat to antibiotic therapy. The high prevalence of bla(CMY) genes and multidrug-resistant genes may also cause therapeutic failure and lack of eradication of these strains by third-generation cephalosporins or cephamycins. PMID- 11874546 TI - PCR-ELISA detection of Escherichia coli in milk. AB - AIMS: The purpose of this study was to develop a reliable molecular procedure for the detection of Escherichia coli in milk. METHODS AND RESULTS: Robust and expeditious DNA extraction and PCR techniques were evaluated using Enzyme-Linked Immunosorbent Assay (ELISA) detection of biotin-labelled amplicons to facilitate optimal detection of E. coli DNA. CONCLUSIONS: It was found that 5 E. coli colony forming units (cfu) could be detected per PCR reaction using the PCR-ELISA system, equating to a sensitivity of detection of 100 E. coli cfu ml(-1) pasteurized milk. SIGNIFICANCE AND IMPACT OF THE STUDY: This approach should facilitate evaluation of milk contamination and enable rapid detection of E. coli mastitis, leading to correct deployment of relevant antibiotic therapy and improved animal welfare. PMID- 11874547 TI - EU Drinking Water Directive reference methods for enumeration of total coliforms and Escherichia coli compared with alternative methods. AB - AIMS: The reference methods for enumeration of total coliforms and Escherichia coli as stated in the European Drinking Water Directive were compared with alternative methods. METHODS AND RESULTS: Laboratories used the reference method on Lactose TTC agar (LTTC), the Colilert/18 system, Laurysulphate Agar (LSA), Chromocult Coliform Agar and the E. coli Direct Plating (DP) method. They enumerated more total coliforms on LTTC than on LSA. CONCLUSIONS: LTTC is suitable for analysis of very clean water samples only, due to heavy background growth. Colilert/18 is a good alternative but it enumerates a broader group of total coliforms, resulting in higher counts. The DP method appeared to be the best choice for enumeration of E. coli because Colilert/18 produces lower counts and false-negative results. SIGNIFICANCE AND IMPACT OF THE STUDY: This study shows the limitations of the EU reference method on LTTC due to lack of selectivity and suggests alternative methods for the enumeration of total coliforms and E. coli. PMID- 11874549 TI - Potentiation of motilin-induced contraction by nitric oxide synthase inhibition in the isolated chicken gastrointestinal tract. AB - The present experiments were designed to determine whether or not endogenous nitric oxide (NO) modifies the contractile response to chicken motilin (ch-MT) in the gastrointestinal (GI) tract (proventriculus and small intestine) of the chicken. ch-MT (1 nmol L(-1)-1 micromol L(-1)) caused contractions of longitudinal muscle strips of the proventriculus through both myogenic and neurogenic (mostly cholinergic) mechanisms. On the other hand, ch-MT (0.1 nmol L( 1)-100 nmol L(-1)) contracted the small intestine (duodenum, jejunum and ileum) only through a myogenic mechanism. L-Nitroarginine methylester (L-NAME) potentiated, and L-arginine inhibited, the ch-MT- induced contraction without affecting the responsiveness of acetylcholine (ACh) or 5-hydroxytryptamine in the proventriculus. Electrical field stimulation (EFS)- and 1,1-dimethyl-4 phenylpiperazinium (DMPP)- induced contractions were also potentiated by L-NAME. The potentiation by L-NAME was prevented by L-arginine but not by D-arginine. However, in the presence of atropine or tetrodotoxin, neither L-NAME nor L arginine modified the responses to ch-MT and DMPP. In contrast to the proventriculus, L-NAME and L-arginine were both ineffective in modifying the ch MT-induced contraction in the small intestine. These results indicate that NO synthase inhibition potentiates the contractile response of ch-MT, EFS and DMPP in the chicken proventriculus through reduction of endogenous NO-mediated presynaptic inhibition on neural ACh release. However, NOS inhibition did not modify the myogenic (direct) action of ch-MT in gastric and intestinal smooth muscles of the chicken. PMID- 11874550 TI - Automated, quantitative analysis of interdigestive small intestinal myoelectric activity in rats. AB - Quantitative analysis of myoelectric activity (EMG), to investigate small intestinal motility in rats, is normally based on manual classification into sequences of phase I, phase II and phase III. This classification is partly subjective. We aimed to develop a more objective method for the analysis of the migrating myoelectric complex (MMC). From the EMG, a derived signal is calculated as a measure of activity. Depending on the level of this derived signal, the EMG is classified into 'quiescent phase', 'irregular phase' or 'activity front'. The threshold levels for these phases are automatically calculated from the EMG data. A proposal for subdivision into MMCs is automatically generated. To calculate MMC length, the user must manually reject nonpropagated activity fronts. While developing the method, more than 19 derived signals were tested. These included variants of spike frequency, signal power and spike-burst length. The spike frequency signal was chosen because it gave minimal deviation from manual classification. Using the new automated method, recordings from the jejunum of 15 healthy rats were analysed (6 h each). The calculated phase lengths were consistent with the results of manual analysis. The presented method allows objective analysis of the interdigestive EMG signals of the small intestine. PMID- 11874551 TI - Antropyloroduodenal, cholecystokinin and feeding responses to pulsatile and non pulsatile intraduodenal lipid infusion. AB - The contribution of the pulsatile nature of gastric emptying to small intestinal feedback mechanisms modulating antropyloroduodenal motility and appetite is unknown. On separate days, eight healthy male volunteers (18-34 years) received randomized, single-blind, intraduodenal (ID) infusions of 10% Intralipid (2 kcal min(-1)), either continuously [CID], or in a pulsatile manner [PID] (5 s on/15 s off) and 0.9% saline (control) administered continuously, each at a rate of 1.8 mL min(-1) for 3 h. During each infusion, subjective ratings of appetite were assessed and antropyloroduodenal pressures recorded with a 16-lumen manometric assembly incorporating a pyloric sleeve sensor. Plasma cholecystokinin was measured from blood collected at regular intervals throughout the infusion. At the end of each infusion the manometric assembly was removed, subjects were offered a buffet meal and the energy and macronutrient content of the meal was measured. Both ID lipid infusions stimulated isolated pyloric pressure waves (IPPWs) (P < 0.001) and basal pyloric pressure (P < 0.01) and suppressed antral (P < 0.05) and duodenal (P < 0.05) pressure waves when compared to controls; there was no difference in the effects of CID and PID lipid on antropyloroduodenal pressures. Infusions of lipid significantly increased plasma CCK concentrations (P < 0.05) compared with saline, but concentrations were not different between the two modes of lipid delivery (P > 0.05, CID vs. PID). Both intraduodenal lipid infusions decreased hunger (P < 0.05), increased fullness (P < 0.05) and reduced energy intake (P < 0.05) when compared with controls; again there was no difference between CID and PID lipid. We conclude that at the infusion rate of similar 2 kcal min(-1), the acute effects of intraduodenal lipid on antropyloroduodenal pressures, plasma CCK concentration and appetite are not modified by a pulsatile mode of lipid delivery into the duodenum. PMID- 11874552 TI - Neuroendocrine control of intestinal mucosal mast cells under physiological conditions. AB - Mast cells are involved in the pathogenesis of both allergies to food and inflammatory bowel disorders. In addition, there are several lines of evidence suggesting that mucosal mast cells also respond to intraluminal stimuli. Our aim was to identify neuroendocrine stimuli that could modify mucosal mast cell activity in the rat. Anaesthetized rats were prepared for duodenal perfusion and mast cell activation was measured by analysis of RMCP II concentration in the duodenal perfusate. Either buffered saline solution or a 5% ovalbumin hydrolysate (OVH) solution was infused into the duodenum. Subdiaphragmatic vagotomy or afferent ablation by intraluminal treatment with capsaicin diminished RMCP II concentration in basal conditions and significantly reduced the response to OVH, which in control animals induced a three-fold increase of the protease. The noradrenergic blockers phentholamine and propranolol significantly diminished RMCP II concentration in basal conditions and completely blocked the response to OVH. Intravenous infusion of cholecystokinin-related peptides also induced a response of mast cells. However, the response was different depending on the peptide. CCK-8 induced a slight increase of RMCP II, whereas both CCK-33 and gastrin induced a significant decrease in mast cell activity. These results show that intraluminal content modulates mucosal mast cell activity by complex mechanisms involving both nervous and endocrine pathway. PMID- 11874553 TI - Morphometric and biomechanical remodelling in the intestine after small bowel resection in the rat. AB - The short-bowel syndrome is a clinical condition caused by intestinal resection. As intestinal adaptation occurs after resection, it can be used as a model for studying morphometric and biomechanical remodelling in the small intestine and to get a better understanding of the pathophysiology of the short-bowel syndrome. The resected rats had a 67% resection of jejunum and ileum. Control animals underwent no operation (nonoperated controls) or an ileal transection with subsequent end-to-end anastomosis (sham-resected controls). The animals were followed for up to 4 weeks after the operation. Changes in biomechanical properties were studied in terms of residual strain (the internal strain remaining when all external loads are removed), opening angle and stress--strain relations referenced to the zero-stress state (the cut-open state where external and internal stresses are released). The resected animals gained less weight than the controls. The intestinal length and diameter increased more in the resected groups than the control groups (P < 0.05), resulting in a larger absorptive surface. Resection induced profound gross morphometric changes and histological alterations characterized by proliferative increases in the tissue layers. The opening angle, along with residual strain at the mucosal and serosal surface, increased in the remnant small intestine (P < 0.05). All changes increased as function of postoperative time and were most prominent in the remnant ileum. However, the stress-strain relationship remained unchanged. In conclusion, this study demonstrated that resection of the majority of the small bowel results in significant remodelling in structural and residual strain properties in the remnant small intestine. The remodelling seems to be guided by the need for a greater absorptive surface area rather than for a change in the stress-strain properties. PMID- 11874554 TI - Effect of bowel cleansing on colonic transit in constipation due to slow transit or evacuation disorder. AB - Colon transit time measurement with radio-opaque markers is a method of studying the passage of luminal contents throughout the colon. Overall colonic transit time (CTT), as well as segmental transit times [right (RTT), left (LTT) and rectosigmoid (RSTT)], can be calculated. We hypothesize that CTT is influenced by faecal impaction when the rectum is emptied infrequently. The aim of this study is to investigate the effect of bowel cleansing on colonic transit time in patients with chronic constipation. In 25 women (age 41 years; range 20-65 years) with constipation according to Thompson criteria, CTT measurement was performed in an unprepared situation and repeated after cleansing with 4 L of Klean Prepreg. Ten healthy female volunteers (age 41 years; range 27-57 years) were used as controls. In constipated patients, CTT decreased from a median 70 h (range 10-130 h) to 48 h (5-94 h) in the cleansed state (P < 0.001). A shortening of transit time was found in all three segments. In 10 patients with slow transit (ST) (CTT > 86 h), CTT decreased from 110 h (range 94-130) to 86 (38-94) (P < 0.001). Five of the 10 patients with ST before bowel cleansing had a CTT below 86 h after cleansing. In female controls, uncleansed CTT and RSTT shortened from 39 h (23 to 62) and 17 h (8-29) to 29 h (17-48) and 10 h (0-20) after bowel cleansing (P=0.058 and P=0.046). Colonic intraluminal contents have a substantial effect on colonic transit. In female controls, bowel cleansing shortened rectosigmoid transit. Women with constipation had faster transit in the cleansed state, however, the distribution of markers was not altered. Despite the effect of bowel cleansing on CTT, it seems unnecessary to prepare the bowel in clinical practice because the differentiation of patients between slow transit constipation and outlet obstruction is not changed. However, because in an infrequent defecation pattern, the influence of faecal impaction is considerable, CTT should be applied with care for critical clinical decisions in the treatment of constipation. PMID- 11874555 TI - Calcium signalling and removal mechanisms in myenteric neurones. AB - To characterize further the Ca2+ signalling mechanisms of myenteric neurones, we studied the effect of thapsigargin, a blocker of the Ca2+-store ATPase, and the mechanisms involved in restoring the intracellular Ca2+ concentration ([Ca2+]i) after activation. Thapsigargin (5 x 10(-6) mol L(-1)) induced an oscillatory [Ca2+]i response in 86.6% of the neurones (n=276), which was blocked by the removal of extracellular Ca2+ and by omega-conotoxin MVIIA (5 x 10(-7) mol L( 1)). The IP3-blocker, 2-aminoethyl-diphenyl-borate (75 x 10(-6) mol L(-1)), blocked or reduced the responses in 74.5% of the neurones. The oscillatory responses induced by the depletion of Ca2+ stores suggest that myenteric neurones might recruite N-type Ca2+ channels as a refill mechanism. Thapsigargin pretreatment increased the amplitude, the upstroke and duration of the K+-induced [Ca2+]i responses. Mitochondrial blockers (rotenone and antimycin/oligomycin) also prolonged the responses, but without affecting the amplitude. Furthermore, it was found that for high [Ca2+]i, the thapsigargin-sensitive Ca2+ uptake was crucial, while mitochondrial blockade affected the Ca2+ uptake over a wide range of concentrations. The Ca2+-sequestering components might also have been compensating for each other, as most drugs only delayed and not inhibited Ca2+ removal. PMID- 11874556 TI - Acute and chronic stress differently affect visceral sensitivity to rectal distension in female rats. AB - Stressful life events are frequently associated with outward signs of irritable bowel syndrome (IBS). Increasing evidence suggests that acute and chronic stress stimuli implicate different physiological mechanisms and neuroendocrine responses. Therefore, we investigated the influence of acute and chronic stress on visceral nociception in female rats and the involvement of colonic mast cells in this effect. The effect of acute and chronic partial restraint stress (PRS) on visceral sensitivity to rectal distension (RD) was assessed by abdominal muscle electromyography. Colonic mast cell activation was determined by measuring histamine release after in vitro stimulation with substance P (SP) in colonic samples from rats experiencing RD vs. controls. Acute PRS significantly enhanced abdominal response to RD compared with sham PRS for all volumes of distension. In contrast, chronic PRS induced a hyperalgesic response for the highest volumes of distension (0.8 and 1.2 mL), but did not affect the number of abdominal contractions for the lowest volume (0.4 mL) compared with controls. Both acute and chronic PRS increased in vitro SP-induced histamine release without affecting mast cell numbers. RD induced similar in vitro histamine release from colonic samples from both acute and chronic PRS rats; this release, however, was significantly higher than that measured in sham-PRS rats. Acute and chronic PRS differently influence visceral sensitivity in response to RD in female rats. This difference, however, cannot be attributed to a different effect of either stress paradigm on mast cell histamine release. PMID- 11874557 TI - Alterations of neurokinin receptors and interstitial cells of Cajal during and after jejunal inflammation induced by Nippostrongylus brasiliensis in the rat. AB - Substance P (SP) and its receptors NK1 and NK2 are widely expressed in the intestinal wall by neurones, interstitial cells of Cajal (ICC) and smooth muscle cells. Changes in SP and/or its NK receptors have been documented during experimental inflammation in animals or inflammatory bowel diseases in humans, but the data concern the acute phase of the inflammatory process. We determined immunohistochemically whether NK receptors and SP were altered in the muscle coat during jejunal inflammation induced by the nematode Nippostrongylus brasiliensis and whether these alterations persisted when inflammation had spontaneously resolved 30 days postinfection. An ultrastructural analysis was also conducted on ICC, nerves and muscle. At day 14, when inflammation peaked, there was a reduction in NK1 receptors in myenteric neurones and in SP-immunoreactive nerve endings. There were also ultrastructural anomalies in synaptic vesicles and NK2 receptor loss in the circular muscle layer. The SP decrease persisted at day 30, whereas neurones and circular muscle cells re-expressed NK1 and NK2 receptors, respectively. The ICC at the deep muscular plexus, located near to the inflammatory site, underwent alterations leading to their complete loss at day 30. These morphological changes are probably associated with impairment in tachykinergic control of jejunal functions leading to the alterations of motility and sensitivity to distension already described in these animals. PMID- 11874558 TI - Low antroduodenal pressure gradients are responsible for gastric emptying of a low-caloric liquid meal in humans. AB - The motor mechanisms responsible for transpyloric flow of gastric contents are still poorly understood. The aim of our study was to investigate the relationship between luminal pressures and gastric wall motion and between gastroduodenal pressure gradients and pressure waves, and ante- and retro-grade transpyloric flow. In eight healthy volunteers, intraluminal pressures were recorded from the antrum and proximal duodenum. Transpyloric flow was monitored simultaneously using duplex ultrasonography, before, during and after ingestion of 300 mL meat soup. Transpyloric emptying occurred as sequences of alternating periods of emptying-reflux-emptying. Approximately one-third of the sequences were not associated with peristalsis. The antroduodenal pressure gradients were significantly lower during nonperistaltic-related emptying than during peristaltic-related emptying (0.15 (0-0.3) kPa, and 1.7 (0.2-2.0) kPa, respectively [mean plus minus (range)], P < 0.005). The duration of emptying episodes not associated with peristalsis were significantly longer than those associated with peristalsis at (6.5 (3-8.7) s and 4.4 (2-6) s, respectively, P=0.059). Manometry detected only 56% of the antral contractions seen on ultrasound. We concluded that gastric emptying of a low-calorie liquid meal occurs both during peristaltic and nonperistaltic antral activity. In spite of lower antroduodenal pressure gradients, the emptying episodes were longer for nonperistaltic emptying, which is likely to be caused by low pyloric resistance. Considerable flow seems to occur without peristalsis during gastric emptying of a low-calorie, liquid meal in humans. PMID- 11874560 TI - Fasciola hepatica cathepsin L suppresses sheep lymphocyte proliferation in vitro and modulates surface CD4 expression on human and ovine T cells. AB - Fasciola hepatica infection has been shown to suppress sheep lymphocyte proliferation in vitro and this is at least partially attributable to excretory/secretory products (ES) released by F. hepatica parasites. We identified a suppressive component in ES by analysing the effect of ES fractions, separated by gel filtration, on the proliferation of sheep T cells in vitro. A major proportion of the suppressive activity in ES was shown to coelute with the cathepsin L proteases: E64, a cysteine protease inhibitor, blocked the suppressive activity of cathepsin L. In order to identify possible mechanisms by which cathepsin L could suppress T cell proliferation, the effect of ES and F. hepatica recombinant cathepsin L (rFhCatL) on the expression of 22 different sheep T cell surface markers was analysed by flow cytometry. Incubation of sheep T cells with ES or two rFhCatL significantly reduced surface CD4 expression and this effect was prevented in the presence of E64. In similar experiments with human T lymphocytes, ES and rFhCatL were shown to down regulate surface CD4 expression. These results show that F. hepatica cathepsin L both suppresses sheep T cell proliferation and reduces surface CD4 expression on both human and ovine T cells. PMID- 11874561 TI - Chemokine and cytokine expression in murine intestinal epithelium following Nippostrongylus brasiliensis infection. AB - Infection of mice with the nematode parasite Nippostrongylus brasiliensis results in a well characterized intestinal mastocytosis with intraepithelial migration of mucosal mast cells (MMC). The molecules mediating this response are unknown. We examined expression of several putative mast cell chemoattractants in intestinal epithelium following N. brasiliensis infection. Expression of the chemokines monocyte chemoattractant protein-1 (MCP-1), macrophage inflammatory protein-1 alpha (MIP-1alpha), RANTES (regulated on activation normal T-cell expressed and secreted), fractalkine, and thymocyte expressed chemokine (TECK); and the cytokines stem cell factor (SCF) and transforming growth factor beta1 (TGFbeta1), was constitutive and no alteration was detected following infection. MCP-1 expression was also constitutive but at much lower levels and increased expression was detected on days 7 and 14 postinfection. Expression of MCP-1 in whole jejunum was at much higher levels than in epithelium. Constitutive expression of MCP-1, MIP-1alpha and TGFbeta1 was also detected in cultured bone marrow-derived homologues of MMC. In an intestinal epithelial cell line (CMT-93), there was constitutive expression of SCF, TGFalpha1, fractalkine and MCP-1. The results show that, in vivo, epithelium is a potentially important source of mast cell chemoattractants. PMID- 11874562 TI - Naturally acquired immunoglobulin (Ig)G subclass antibodies to crude asexual Plasmodium falciparum lysates: evidence for association with protection for IgG1 and disease for IgG2. AB - There is longstanding evidence for a role of immunoglobulin (Ig)G in protection against malarial disease and infection. IgG1 and IgG3 have been shown to be particularly efficient at associating with monocytes in potentially protective mechanisms (i.e. antibody-dependent cellular inhibition, opsonization and phagocytosis). Conversely, there is some evidence that IgG2 (and possibly IgG4) antibodies may be antagonistic to this protection. The protective effect of IgG subclass antibody activity present before the beginning of a malaria transmission season (preseason antibody levels) against severe malaria has not been tested in longitudinal studies. We measured IgG class and subclass antibody levels specific to crude Plasmodium falciparum lysates by enzyme linked immunosorbent assay in a case-control study of 76 children on the coast of Kenya. The mean optical density values for both IgG class and subclass antibodies were not significantly different between the children who developed severe malaria and those who remained healthy during an observation period of two malaria transmission seasons. However, elevated levels of IgG1 in relation to levels of IgG2 and IgG4 antibodies were associated with protection from severe malaria (P = 0.02). Conversely, elevated levels of IgG2 in relation to IgG1 and IgG3 antibodies were associated with a higher risk of developing severe malaria (P = 0.006). PMID- 11874563 TI - A comparison of cellular and humoral immune responses to trichuroid derived antigens in human trichuriasis. AB - Individuals, residing in a region highly endemic for Trichuris trichiura, were examined for cytokine and proliferative responses to T. trichiura worm homogenate (TtAg), T. trichiura excretory/secretory products (TtES) and the equivalent antigenic preparations from the murine whipworm, Trichuris muris. Serum antibody levels against TtAg, T. muris worm homogenate and T. muris ES products were also studied. Measurable levels of immunoglobulin (Ig)G1, IgG4, IgA and IgE against T. muris antigens were detected, indicating a degree of conservation of epitopes between antigens derived from both species. Although levels of interleukin (IL) 4, IL-10, IL-13, tumour necrosis factor (TNF)-alpha and proliferative responses produced were comparable between homogenate antigens of either species and ES antigens of either species, a markedly different cellular response was observed in cultures stimulated with homogenate antigens compared to ES antigens. ES antigens preferentially induced IL-10 (P > 0.001) and TNF-alpha (P > 0.001) production, whereas levels of IL-4 (P > 0.001), IL-13 (P > 0.001) and proliferative responses (P > 0.001) were greater in cultures stimulated with whole worm extracts. Our findings suggest that T. muris preparations could be used as an alternative to T. trichiura proteins as a source of antigens in ex vivo cultures and that ES products stimulate a distinct immune response compared to somatic antigens. PMID- 11874564 TI - The role of B cells in immunity against larval Strongyloides stercoralis in mice. AB - The objective of this study was to examine the role of B cells in primary and challenge infections of larval Strongyloides stercoralis in mice. Two strains of B-cell deficient mice were used in these studies, microMT mice that lack all B cells and Xid mice that lack B-1 cells. Primary immune responses in microMT mice were sufficient to eliminate all parasites within 1 week after infection. Immunized microMT and Xid mice, however, were unable to kill challenge parasites at 24 h post infection, the time that they were eliminated in immunized wild-type mice. This was despite having a significant increase in interleukin-5 secreting cells and high numbers of eosinophils in the microenvironment of the challenge larvae. In addition, immunized Xid mice did not generate parasite-specific immunoglobulin (Ig)M but did develop a weak IgG response compared to wild-type mice. These results demonstrate a dichotomy in the requirement of B cells in immunity to S. stercoralis. B cells are not required in the primary response, yet they are required in the secondary immune response. B-1 cells are required for the secondary immune response and their role appears to be the production of IgM and not as a source of immunoregulatory molecules. PMID- 11874566 TI - Antigenemia at 10 years after diethylcarbamazine treatment of asymptomatic microfilaraemic individuals: marginal conversion to infection-free state. AB - A group of asymptomatic microfilaraemic individuals (n = 44, 29 males, 15 females) living in a Wuchereria bancrofti endemic region of Orissa, India, was treated with a standard regimen of diethylcarbamazine (12 days, 6 mg/kg) in 1990. The incidence of microfilaraemia and antigenemia (Og4C3) was determined after a gap of 10 years in 2000. Nineteen individuals reacquired microfilariae (43.2%, 11 males, eight females), five males developed hydrocele, two females became acute filarial patients and 18 subjects (13 males, five females) were asymptomatic amicrofilaraemics. Filarial antigen was detected in 36 individuals (81.2%, 27 male, nine female) comprising microfilaraemics, amicrofilaraemics and diseased. Only eight individuals (18.2%, two males, six females) remained antigen free. PMID- 11874565 TI - Passive protection against fasciolosis in mice by immunization with a monoclonal antibody (ES-78 MoAb). AB - Recently, we reported a partial characterization of the epitope recognized by the ES-78 monoclonal antibody (MoAb). This monoclonal antibody was obtained from spleen lymphocytes of a mouse immunized with excretory-secretory antigens of Fasciola hepatica adult worms. In the present study, we report the results obtained in experiments of passive protection using this MoAb in BALB/c mice infected with 15 Fasciola hepatica metacercariae. The monoclonal antibody was able to reduce the parasite burden when administered 24 h before challenge but not when delivered 7 days after challenge. The antibody recognition of digestive tract structures in 3-week-old parasites was demonstrated by immune histochemical techniques. The antigens purified by affinity chromatography using this antibody had molecular weights of 14-20, 25-29 and 36-45 kDa and demonstrated proteinase activity similar to cathepsin L. These results suggest that the antigens carrying the epitope recognized by the ES-78 MoAb may be used as target in the protection against fasciolosis. PMID- 11874567 TI - Double mutation cpSRP43--/cpSRP54-- is necessary to abolish the cpSRP pathway required for thylakoid targeting of the light-harvesting chlorophyll proteins. AB - Biochemical and genetic studies have established that the light-harvesting chlorophyll proteins (LHCPs) of the photosystems use the cpSRP (chloroplast signal recognition particle) pathway for their targeting to thylakoids. Previous analyses of single cpSRP mutants, chaos and ffc, deficient in cpSRP43 and cpSRP54, respectively, have revealed that half of the LHCPs are still integrated into the thylakoid membranes. Surprisingly, the effects of both mutations are additive in the double mutant ffc/chaos described here. This mutant has pale yellow leaves at all stages of growth and drastically reduced levels of all the LHCPs except Lhcb 4. Although the chloroplasts have a normal shape, the thylakoid structure is affected by the mutation, probably as a consequence of reduction of all the LHCPs. ELIPs (early light-inducible proteins), nuclear-encoded proteins related to the LHCP family and inducible by light stress, were also drastically reduced in the double mutant. However, proteins targeted by other chloroplastic targeting pathways (DeltapH, Sec and spontaneous pathways) accumulated to similar levels in the wild-type and the double mutant. Therefore, the near total loss of LHCPs and ELIPs in the double mutant suggests that cpSRP is the predominant, if not exclusive, targeting pathway for these proteins. Phenotypic analysis of the double mutant, compared to the single mutants, suggests that the cpSRP subunits cpSRP43 and cpSRP54 contribute to antenna targeting in an independent but additive way. PMID- 11874568 TI - A peptide methionine sulfoxide reductase highly expressed in photosynthetic tissue in Arabidopsis thaliana can protect the chaperone-like activity of a chloroplast-localized small heat shock protein. AB - The oxidation of methionine residues in proteins to methionine sulfoxides occurs frequently and protein repair by reduction of the methionine sulfoxides is mediated by an enzyme, peptide methionine sulfoxide reductase (PMSR, EC 1.8.4.6), universally present in the genomes of all so far sequenced organisms. Recently, five PMSR-like genes were identified in Arabidopsis thaliana, including one plastidic isoform, chloroplast localised plastidial peptide methionine sulfoxide reductase (pPMSR) that was chloroplast-localized and highly expressed in actively photosynthesizing tissue (Sadanandom A et al., 2000). However, no endogenous substrate to the pPMSR was identified. Here we report that a set of highly conserved methionine residues in Hsp21, a chloroplast-localized small heat shock protein, can become sulfoxidized and thereafter reduced back to methionines by this pPMSR. The pPMSR activity was evaluated using recombinantly expressed pPMSR and Hsp21 from Arabidopsis thaliana and a direct detection of methionine sulfoxides in Hsp21 by mass spectrometry. The pPMSR-catalyzed reduction of Hsp21 methionine sulfoxides occurred on a minute time-scale, was ultimately DTT dependent and led to recovery of Hsp21 conformation and chaperone-like activity, both of which are lost upon methionine sulfoxidation (Harndahl et al., 2001). These data indicate that one important function of pPMSR may be to prevent inactivation of Hsp21 by methionine sulfoxidation, since small heat shock proteins are crucial for cellular resistance to oxidative stress. PMID- 11874569 TI - Identification, subcellular localization and some properties of a cysteine-rich suppressor of gene silencing encoded by peanut clump virus. AB - In plants, post-transcriptional gene silencing (PTGS) is part of a defence mechanism against virus infection. Several plant viruses have been shown to encode proteins which can counteract PTGS. In this paper it is demonstrated that P15 of peanut clump pecluvirus (PCV) has anti-PTGS activity. P15 is a small cysteine-rich protein with no sequence similarity to previously described PTGS suppressor proteins which has several novel properties. It possesses four C terminal proximal heptad repeats that can potentially mediate a coiled-coil interaction and is targeted to peroxisomes via a C-terminal SKL motif. The coiled coil sequence is necessary for the anti-PTGS activity of P15, but the peroxisomal localization signal is not, although it is required for efficient intercellular movement of the virus. PMID- 11874570 TI - An EDS1 orthologue is required for N-mediated resistance against tobacco mosaic virus. AB - In Arabidopsis, EDS1 is essential for disease resistance conferred by a structural subset of resistance (R) proteins containing a nucleotide-binding site, leucine-rich-repeats and amino-terminal similarity to animal Toll and Interleukin-1 (so-called TIR-NBS-LRR proteins). EDS1 is not required by NBS-LRR proteins that possess an amino-terminal coiled-coil motif (CC-NBS-LRR proteins). Using virus-induced gene silencing (VIGS) of a Nicotiana benthaminana EDS1 orthologue, we investigated the role of EDS1 in resistance specified by structurally distinct R genes in transgenic N. benthamiana. Resistance against tobacco mosaic virus mediated by tobacco N, a TIR-NBS-LRR protein, was EDS1 dependent. Two other R proteins, Pto (a protein kinase), and Rx (a CC-NBS-LRR protein) recognizing, respectively, a bacterial and viral pathogen did not require EDS1. These data, together with the finding that expression of N. benthamiana and Arabidopsis EDS1 mRNAs are similarly regulated, lead us to conclude that recruitment of EDS1 by TIR-NBS-LRR proteins is evolutionarily conserved between dicotyledenous plant species in resistance against bacterial, oomycete and viral pathogens. We further demonstrate that VIGS is a useful approach to dissect resistance signaling pathways in a genetically intractable plant species. PMID- 11874571 TI - A novel extinction screen in Arabidopsis thaliana identifies mutant plants defective in early microsporangial development. AB - Few Arabidopsis mutants defective in early male or female germline development have been reported. A novel extinction screen has been devised which permits the identification of mutants deficient in the earliest stages of anther development. Using mutagenized plants carrying GUS reporter constructs driven by tapetal specific promoters originally derived from Brassica genes, a wide spectrum of mutants have been identified in Arabidopsis, ranging from those defective in archesporial cell differentiation to others expressed later in development. Crosses between these lines and known anther development mutants have enabled the identification of lines carrying mutations in genes expressed during very early anther formation. Initial characterization reveals these early mutants fall into two classes, gne (GUS-negative) 1-like, and gne2-like. Members of the gne1 mutant class initiate all four layers of the anther wall and an appropriate number of sporogenous cells; however, as development proceeds the tapetal and middle-layer cells enlarge, eventually crushing the sporogenous cells. The gne2 class anthers are disrupted at an earlier stage, with the middle and tapetal layers failing to form, and an excess of sporogenous cells developing until the germline aborts late in meiosis II. Analysis of these mutants has already raised questions about the accuracy of current models of angiosperm anther development. PMID- 11874572 TI - Fusion genetic analysis of jasmonate-signalling mutants in Arabidopsis. AB - Jasmonates induce plant-defence responses and act to regulate defence-related genes including positive feedback of the lipoxygenase 2 (LOX2) gene involved in jasmonate synthesis. To identify jasmonate-signalling mutants, we used a fusion genetic strategy in which the firefly luciferase (FLUC) and Escherichia coli beta glucuronidase (GUS) reporters were expressed under control of the jasmonate responsive LOX2 promoter. Spatial and temporal patterns of reporter expression were determined initially, and revealed that JA-responsive expression from the LOX2 promoter required de novo protein synthesis. Reporter activity was also induced by the protein kinase inhibitor staurosporine and antagonized by the protein phosphatase inhibitor okadaic acid. FLUC bio-imaging, RNA gel-blot analysis and progeny analyses identified three recessive mutants that underexpress the FLUC reporter, designated jue1, 2 and 3, as well as two recessive mutants, designated joe1 and 2, that overexpress the reporter. Genetic analysis indicated that reporter overexpression in the joe mutants requires COI. joe1 responded to MeJA with increased anthocyanin accumulation, while joe2 responded with decreased root growth inhibition. In addition, reporter induction and endogenous LOX2 expression by staurosporine was absent in joe2. PMID- 11874573 TI - Restoration of stamen development and production of functional pollen in an alloplasmic CMS tobacco line by ectopic expression of the Arabidopsis thaliana SUPERMAN gene. AB - The alloplasmic male-sterile tobacco line Nta(rep)S, combining the nucleus of Nicotiana tabacum with the cytoplasm of Nicotiana repanda, exhibits cadastral type anomalies due to a fusion of several stamens with the pistil. These anomalies share similarities with Arabidopsis superman mutants. SUPERMAN (SUP) is a cadastral gene controlling the boundary between whorls 3 (androecium) and 4 (gynoecium). Thus we hypothesized that the expression of the tobacco SUP orthologue might be impaired in the alloplasmic Nta(rep)S line, and that the deficiency could be complemented by the Arabidopsis SUP gene. Here we show that the ectopic expression of SUP in the alloplasmic male-sterile tobacco line Nta(rep)S significantly increases the frequency of flowers possessing free stamens, inducing the recovery of a proper structure for whorls 3 and 4. Furthermore, flowers of transgenic plants show a significant improvement of the morphology of stamens, and more particularly of the anthers, which are able to produce few but functional pollen. The data show that ectopic expression of Arabidopsis SUP reactivates the regulatory cascade of anther development. The plausible causes of the developmental defects of anthers in the alloplasmic male sterile tobacco line are discussed in relation to the model of regulation of the Arabidopsis SUP gene. PMID- 11874574 TI - Cell cycle-dependent regulation of telomerase activity by auxin, abscisic acid and protein phosphorylation in tobacco BY-2 suspension culture cells. AB - Telomerase is a specialized RNA-directed DNA polymerase that adds telomeric repeats onto the ends of linear eukaryotic chromosomes. It was recently reported that the low, basal level of telomerase activity markedly increased at early S phase of the cell cycle, and auxin further increased the S-phase-specific telomerase activity in tobacco BY-2 cells. In this study we show that abscisic acid (ABA), a phytohormone known to induce the cyclin-dependent protein kinase inhibitor, effectively abolished both the auxin- and S-phase-specific activation of telomerase in a concentration- and time-dependent fashion in synchronized tobacco BY-2 cells. These results suggest that there exists a hormonal cross-talk between auxin and ABA for the regulation of telomerase activity during the cell cycle of tobacco cells. Treatment of synchronized BY-2 cells with the protein kinase inhibitor staurosporine or H-7 effectively prevented the S-phase-specific activation of telomerase activity. By contrast, when okadaic acid or cantharidin, potent inhibitors of protein phosphatase 2A (PP2A), was applied to the cells, the S-phase-specific high level of telomerase activity was continuously maintained in the cell cycle for at least 14 h after release from M-phase arrest. Incubation of tobacco cell extracts with exogenous PP2A rapidly abrogated in vitro telomerase activity, while okadaic acid and cantharidin blocked the action of PP2A, effectively restoring in vitro telomerase activity. Taken together, these findings are discussed in the light of the suggestion that antagonistic functions of auxin and ABA, and reciprocal phosphorylation and dephosphorylation of telomerase complex, are necessarily involved in the cell cycle-dependent modulation of telomerase activity in tobacco cells. PMID- 11874575 TI - Cultivated tomato has defects in both S-RNase and HT genes required for stylar function of self-incompatibility. AB - Cultivated tomato (Lycopersicon esculentum), a self-compatible species, evolved from self-incompatible (SI) species in the genus Lycopersicon following a breakdown of the self-incompatibility system. In order to elucidate the molecular basis of this breakdown in L. esculentum, we first analysed the stylar proteins with an in-gel assay for ribonuclease activity and 2D-PAGE. No S-RNase protein or its activity was detected in the style of L. esculentum. We then introduced the S6-RNase gene from an SI relative, L. peruvianum, into L. esculentum. However, the styles of transgenic plants expressing S6-RNase at levels comparable to those found in the L. peruvianum style were unable to reject self-pollen and L. peruvianum pollen in an allele-specific manner. This indicated that defect in the S-RNase expression was not the sole reason for the loss of self-incompatibility in tomato. The asparagine-rich HT protein, originally identified from the style of Nicotiana alata, is the other stylar factor involved in self-incompatibility reaction. We cloned and sequenced two distinct genes encoding HT-A and HT-B proteins from L. peruvianum (LpHT-A and LpHT-B) and L. esculentum (LeHT-A and LeHT-B). A frame shift mutation in the coding sequence of LeHT-A and a stop codon in the ORF of LeHT-B were found, and no LeHT-B transcript was detected in the style of L. esculentum. The results suggest that the breakdown of self incompatibility in cultivated tomato is associated with loss-of-function mutations in both S-RNase and HT genes. PMID- 11874576 TI - Activation of AtMEK1, an Arabidopsis mitogen-activated protein kinase kinase, in vitro and in vivo: analysis of active mutants expressed in E. coli and generation of the active form in stress response in seedlings. AB - The mitogen-activated protein kinase (MAPK) cascade, consisting of MAPK, MAPK kinase (MAPKK) and MAPK kinase kinase (MAPKKK), is the signaling system that relays various external signals, including mitogens and stresses in eukaryotes. MAPKK is activated by phosphorylation in the consensus motif, SXXXS/T, in animals, but the regulation mechanism for the plant MAPKK by phosphorylation, having the putative phosphorylation motif of S/TXXXXXS/T, is not yet fully clarified. Here we constructed a series of mutants of AtMEK1, an Arabidopsis MAPKK, having the sequence T218-X-S220-X-X-X-S224 that fits both of the plant- and animal-type motifs. We show that the two double-mutant proteins replacing Thr 218/Ser-224 and Ser-220/Ser-224 by Glu expressed in Escherichia coli show a constitutive activity to phosphorylate the Thr and Tyr residues of the kinase negative mutant of an Arabidopsis MAPK, named ATMPK4, in vitro. The mutation analysis of AtMEK1 replacing Thr-218 and Ser-220 to Ala suggested that Thr-218 is autophosphorylated by the enzyme. The wild-type ATMPK4 was also phosphorylated by the active mutants of AtMEK1 and showed a high protein kinase activity toward myelin basic proteins. In contrast, ATMPK3, another Arabidopsis MAPK, was a poor substrate of this plant MAPKK, indicating that AtMEK1 has a substrate specificity preferring ATMPK4 to ATMPK3, at least in vitro. Furthermore, AtMEK1 immunoprecipitated from Arabidopsis seedlings stimulated with wounding, cold, drought, and high salt showed an elevated protein kinase activity toward the kinase-negative ATMPK4, while the amounts of the AtMEK1 protein did not change significantly. These data indicate that the AtMEK1 becomes an active form through phosphorylation and activates its downstream target ATMPK4 in stress response in Arabidopsis. PMID- 11874577 TI - Salt causes ion disequilibrium-induced programmed cell death in yeast and plants. AB - Programmed cell death (PCD) is a fundamental cellular process conserved in metazoans, plants and yeast. Evidence is presented that salt induces PCD in yeast and plants because of an ionic, rather than osmotic, etiology. In yeast, NaCl inhibited growth and caused a time-dependent reduction in viability that was preceded by DNA fragmentation. NaCl also induced the cytological hallmarks of lysigenous-type PCD, including nuclear fragmentation, vacuolation and lysis. The human anti-apoptotic protein Bcl-2 increased salt tolerance of wild-type yeast strain and calcineurin-deficient yeast mutant (cnb1Delta) that is defective for ion homeostasis, but had no effect on the NaCl or sorbitol sensitivity of the osmotic hypersensitive hog1Delta mutant -- results that further link PCD in the response to the ion disequilibrium under salt stress. Bcl-2 suppression of cnb1Delta salt sensitivity was ENA1 (P-type ATPase gene)-dependent, due in part to transcriptional activation. Salt-induced PCD (TUNEL staining and DNA laddering) in primary roots of both Arabidopsis thaliana wild type (Col-1 gl1) and sos1 (salt overly sensitive) mutant seedlings correlated positively with treatment lethality. Wild-type plants survived salt stress levels that were lethal to sos1 plants because secondary roots were produced from the shoot/root transition zone. PCD-mediated elimination of the primary root in response to salt shock appears to be an adaptive mechanism that facilitates the production of roots more able to cope with a saline environment. Both salt-sensitive mutants of yeast (cnb1Delta) and Arabidopsis (sos1) exhibit substantially more profound PCD symptoms, indicating that salt-induced PCD is mediated by ion disequilibrium. PMID- 11874578 TI - Redistribution of membrane proteins between the Golgi apparatus and endoplasmic reticulum in plants is reversible and not dependent on cytoskeletal networks. AB - We have fused the signal anchor sequences of a rat sialyl transferase and a human galactosyl transferase along with the Arabidopsis homologue of the yeast HDEL receptor (AtERD2) to the jellyfish green fluorescent protein (GFP) and transiently expressed the chimeric genes in tobacco leaves. All constructs targeted the Golgi apparatus and co-expression with DsRed fusions along with immunolabelling of stably transformed BY2 cells indicated that the fusion proteins located all Golgi stacks. Exposure of tissue to brefeldin A (BFA) resulted in the reversible redistribution of ST-GFP into the endoplasmic reticulum. This effect occurred in the presence of a protein synthesis inhibitor and also in the absence of microtubules or actin filaments. Likewise, reformation of Golgi stacks on removal of BFA was not dependent on either protein synthesis or the cytoskeleton. These data suggest that ER to Golgi transport in the cell types observed does not require cytoskeletal-based mechanochemical motor systems. However, expression of an inhibitory mutant of Arabidopsis Rab 1b (AtRab1b(N121I) significantly slowed down the recovery of Golgi fluorescence in BFA treated cells indicating a role for Rab1 in regulating ER to Golgi anterograde transport. PMID- 11874580 TI - Dialysis access failure: an indication for immediate kidney transplantation. AB - In the United States, presently there are more than 50,000 patients with end stage renal disease (ESRD) awaiting a cadaveric kidney and each year less than a quarter receive kidney transplantation. Although the real incidence in unknown, a significant number of these patients die due to lack of dialysis access. While various medical necessities are indications for emergent transplantation of other organs, the current kidney allocation system of the United Network for Organ Sharing (UNOS) makes no room for those ESRD patients whose death is imminent due to lack of vascular access. Local organ procurement organizations (OPOs) are allowed to make decisions based on arbitrary policies (or no policies at all) which often falter and fail to deal appropriately with a largely ignored issue. The growing wait for cadaveric kidneys makes the problem of ESRD patients dying due to lack of dialysis access increasingly important and one that must be addressed through a revision of the UNOS kidney allocation system. PMID- 11874581 TI - Dissociation between dialysis adequacy and Kt/V. AB - Since the initiation of dialysis, nephrologists have sought an index (or indices) for the adequacy of toxic solute removal. This quest has been characterized by a gradual shift in thinking, ending with a preference for dynamic parameters such as clearances normalized for body size (Kt/V). The threshold Kt/V, however, has changed over the years. While present guidelines suggest 1.2 with single-pool kinetics, higher levels might be proposed in the future. In spite of the known relation between Kt/V and survival, the accuracy of this parameter as a representative of the removal of the whole spectrum of compounds that are responsible for uremia is problematic. Kt/V only assesses the removal of a water soluble compound from the body water through mostly hydrophilic membranes to the dialysate water. Furthermore, the small size of urea means that convective and/or diffusive transfer through a given semipermeable membrane is unlikely to be representative of larger molecules, especially if dialyzers with a small pore size are applied. Urea kinetics are also poorly representative of the removal of small protein-bound molecules and intracellular solutes with cell membrane limited clearance. Finally, it should be realized that the Kt/V concept has been developed in a specific population, that is, a group of renal failure patients with few comorbidities, submitted to short intermittent hemodialysis with small pore bioincompatible membranes very likely using dialysate of lower quality than that used today. Kt/V might well become less accurate and useful in predicting outcomes as different dialysis conditions are pursued, such as dialysis with biocompatible and/or large-pore membranes, (ultra) pure dialysate, alternative time frames, high levels of convection, and/or in populations with a different distribution of body mass. PMID- 11874582 TI - Should hemoglobin be normalized in patients with chronic kidney disease? AB - In the last decade the nephrology community has learned much about the impact of anemia on patients with kidney disease. Therapy of anemia can correct many of the symptoms which seriously compromise patient function. Despite the obvious benefits, controversy continues regarding the optimal target hemoglobin concentration both in patients prior to dialysis and in dialysis populations. In this editorial we review the clinical data that contribute to this controversy and the physiologic concepts underlying the treatment of anemia. Furthermore, we discuss the need to individualize hemoglobin targets for specific patient populations and the importance of early identification and treatment of anemia in patients with kidney disease. The economic impact of normalizing hemoglobin with the use of erythropoietin and intravenous or oral iron has affected clinical practice over the last decade. Current guidelines published by Kidney Disease Outcomes and Quality Initiative (KDOQI), the European Working Group on Anemia Management, and the Canadian Society of Nephrology all recommend target hemoglobin concentrations and thresholds for initiation of therapy and also suggest the need for reevaluation of current targets in light of new evidence. This editorial supports those guidelines and challenges the reader to critically evaluate current practice in the context of the accumulating data and the physiologic principles discussed herein. The therapy of anemia in patients with chronic kidney disease (CKD) is becoming increasingly sophisticated and is an essential component of care in patients with CKD. However, the effects of therapy will be most impressive when accompanied by the optimal care of all hemodynamic and metabolic abnormalities that are associated with CKD. PMID- 11874583 TI - Do AV fistulas contribute to cardiac mortality in hemodialysis patients? AB - Potential fistula-related problems which may impact on patient survival include high fistula flow with hyperkinetic circulation and cardiac failure, low fistula flow with the risks of underdialysis and fistula thrombosis, vascular access infection with local or systemic manifestations, and possibly induction and maintenance of a microinflammatory state (at least for synthetic grafts). All these complications are much more common with prosthetic grafts than with native arteriovenous (AV) fistulas. Fistula flow should be monitored (e.g., by duplex sonography) to guarantee adequate flow and permit preemptive intervention to avoid fistula thrombosis. PMID- 11874584 TI - How can the cardiac death rate be reduced in dialysis patients? PMID- 11874586 TI - How can the cardiac death rate be reduced in dialysis patients? PMID- 11874585 TI - How can the cardiac death rate be reduced in dialysis patients? PMID- 11874587 TI - How can the cardiac death rate be reduced in dialysis patients? PMID- 11874588 TI - How can the cardiac death rate be reduced in dialysis patients? PMID- 11874589 TI - Quality outcomes and obstacles to their achievement in end-stage renal disease. AB - Much effort has gone into improving the quality of care in end-stage renal disease (ESRD) in recent years. In particular, the National Kidney Foundation's Kidney Disease Outcomes Quality Initiative (KDOQI) serves as a positive example. However, there are certain barriers to quality of care that may hinder such efforts. In this article we discuss obstacles to quality of care in ESRD and present potential solutions. PMID- 11874590 TI - Improving hemodialysis patient outcomes: a step-by-step approach. AB - Hemodialysis patient outcomes are often suboptimal, with one-sixth of patients receiving an inadequate dialysis dose, one-fourth using catheters for vascular access, and more than one-half being malnourished. This review describes a four step approach for improving dialysis patient outcomes. First, select an outcome to improve. This can be a global outcome such as mortality, morbidity, quality of life, or health care costs or an intermediate outcome (such as dialysis dose) that has a demonstrated link with a global outcome. Second, determine barriers to optimal outcomes. Both patient factors (such as noncompliance) and provider factors (such as the process of care) may act as barriers. Third, intervene on specific barriers. Interventions may involve providing medical or surgical treatment, changing patient or provider behavior, or modifying the system of care. Fourth, disseminate the intervention to other settings. "Early adopters" (those who quickly adopt an innovation) and "opinion leaders" (well-respected local physicians) are especially influential in determining the rate of dissemination. By using this approach, physicians can move from helping an individual patient to improving the outcomes of many patients. PMID- 11874591 TI - Utilizing a disease management approach to improve ESRD patient outcomes. AB - In this era of processes and systems to improve quality, disease management is one methodology to improve care delivery and outcomes for patients with chronic kidney disease (CKD). In most disease management systems a senior renal nurse coordinates all aspects of the patient's care and ensures that the prescribed and necessary care is delivered for both CKD-related and comorbid conditions. The nurse also continually monitors outcomes on quality indicators and key performance measures. These outcome data are then aggregated and analyzed, are compared with local and national benchmarks, and drive the continuous quality improvement (CQI) process. Such a system attempts to centralize the currently fragmented care delivery system, continually improve patient outcomes, and conserve scarce economic resources. Early data suggest a disease management approach may improve both the morbidity and mortality of CKD patients. PMID- 11874592 TI - Quality end-of-life care in dialysis units. AB - Quality end-of-life care has not been a priority in dialysis units and patients often experience prolonged dying while suffering needlessly. Advance directives (ADs) and decisions to stop dialysis have been highlighted by the medical profession as priorities in improving the quality of care, yet these are only two aspects of end-of-life care. They may not reflect patients' priorities and may not have the expected impact in improving the quality of end-of-life care. This review argues that quality end-of-life care should be a clinical priority in the care of dialysis patients; end-of-life care needs to be developed primarily from the patients' perspective; a clinical framework is required that integrates many aspects of end-of-life care; and end-of-life care should be initiated much earlier in the course of patients' illnesses than traditionally is done. By communicating more effectively and sooner with patients, their values and needs can be identified so we will be better able to plan and facilitate their end-of life care and improve their experience of dying. PMID- 11874593 TI - The clinical and economic impact of pharmaceutical care in end-stage renal disease patients. AB - End-stage renal disease (ESRD) patients are medically complex, require multiple medications for treatments of their various comorbidities, and cost the healthcare system billions of dollars each year. These patients are at risk of drug-related problems (DRPs) that may lead to increased morbidity, mortality, and cost to the healthcare system. Review of the literature demonstrates that pharmaceutical care provided by pharmacists improves ESRD patient care. Pharmacist review of ESRD patients' medication profiles and medical records has shown to be beneficial in identifying and resolving DRPs. Economic analysis suggests that for every $1 spent on pharmaceutical care, the healthcare system saves an estimated $3.98. Provision of pharmaceutical care by pharmacists should be consdiered for all ESRD patients. PMID- 11874594 TI - Water treatment for hemodialysis: ensuring patient safety. AB - Patient safety has become an important focus of the Institute of Medicine and the medical community. Although hemodialysis is a routine therapy, it is nonetheless a complex procedure where errors can occur. In particular, errors related to water quality can lead to patient injury and to increased medical costs. Using the Institute of Medicine report on errors in medicine as a basis, this article discusses previously published incidents of patient injury related to water quality in terms of the types of errors that occurred. Epidemiologic techniques provide a framework to identify, correct, and possibly avert these types of errors in the future. While the ultimate responsibility for ensuring water quality rests with the medical director of the hemodialysis unit, patient safety should be a concern of all members of the nephrology community. PMID- 11874595 TI - Herbs, menopause, and dialysis. AB - Women with chronic kidney disease (CKD) are at increased risk for menstrual disorders, early menopause, and osteoporosis, and rarely discuss gynecologic and reproductive issues with their nephrologist. Various complementary and alternative medicine (CAM) products are of interest to women with end-stage renal disease (ESRD) who have these disorders. However, very little is known about the specifics of using herbal medicines in patients on chronic dialysis, resulting in numerous problems when patients and providers try to ascertain the safety and efficacy of these products. This article reviews evidence regarding the safety and efficacy of black cohosh, ginseng, chastetree, dong quai, evening primrose oil, soy products, and the so-called natural hormones. Pharmacologic parameters important to evaluating the quality of botanical products are discussed, along with recommendations and information resources. PMID- 11874596 TI - An introduction to epidemiology and biostatistics and issues in interpretation of studies. AB - Epidemiology is the basic medical science that focuses on the distribution and determinants of disease frequency in human populations. An understanding of the tools of epidemiology is helpful in defining the limitations of medical research and evaluating the conclusions of studies. This is the first in a series of three articles whose objective will be to present the basic concepts of epidemiology and biostatistics. Examples of each of the tools and limitations discussed from studies of patients with end-stage renal disease (ESRD) will be presented to provide the reader with a practical application of the concepts. This series of articles will help the reader to weigh methods and study designs to understand the appropriate conclusions that may be drawn from any data. PMID- 11874597 TI - Cardiac troponins: utility in renal insufficiency and end-stage renal disease. AB - Currently available serum markers of cardiac injury in patients with renal insufficiency suffer from impaired sensitivity and specificity. Cardiac troponins (cTnI, cTnT) are relatively new diagnostic markers of myocardial injury and have gained widespread application in the non-renal-failure population to diagnose myocardial infarction. Over the past few years the specificity and sensitivity of cardiac troponins for diagnosing acute myocardial infarction in patients with renal dysfunction have been examined. Most data indicate that cardiac troponin I has an excellent specificity, but until more studies are available this marker should be considered a useful but imperfect serum marker of an acute coronary syndrome in patients with underlying renal dysfunction. PMID- 11874598 TI - Safety of oral carnitine in dialysis patients. PMID- 11874599 TI - Alteplase use for clotted catheters. PMID- 11874600 TI - PTH assays in renal failure. PMID- 11874601 TI - Lower target calcium levels in dialysis patients. PMID- 11874603 TI - The Respiratory Care journal conferences: 20 years of excellence and innovation. PMID- 11874604 TI - Invasive mechanical ventilation in adults:implementation, management, weaning, & follow-up. PMID- 11874605 TI - Indications for mechanical ventilation in adults with acute respiratory failure. AB - Increased understanding of the mechanisms and effects of acute respiratory failure has not been accompanied by more precise criteria by which the clinician can determine when intubation should be carried out and invasive positive pressure ventilation (IPPV) instituted in a given patient. The indications traditionally offered in reviews and textbooks have tended to be either so broad as not to be very helpful in an individual case, or of questionable clinical relevance and too cumbersome for practical use. This review updates the indications for IPPV in adult patients with acute respiratory failure by examining available evidence from clinical trials and by considering new management alternatives that have become available in the last 20 years. Indications for IPPV based on specific threshold values for P(CO2) and pH or on various indices of arterial oxygenation have generally not been validated by clinical evidence, and it is unlikely that any cutoff value would be applicable to all patients or all categories of acute respiratory failure. Stated another way, there is probably no single value for arterial P(CO2), pH, or P(O2) that by itself constitutes an indication for IPPV. Compelling face validity justifies the use of IPPV in cases of apnea or when it appears certain that respiratory arrest is about to occur. However, dyspnea, tachypnea, or the subjective impression of respiratory distress are probably not in themselves justification for emergency intubation. It should be possible to avoid IPPV and its attendant complications in many cases of acute hypercapnic respiratory failure. In acute exacerbations of chronic obstructive pulmonary disease, noninvasive positive-pressure ventilation (NPPV) should be the initial ventilation approach unless the patient has one of several specific exclusion criteria such as cardiovascular instability or severely impaired mental status. It may also be possible to avoid intubation through the use of NPPV in certain immunocompromised patients with early acute hypoxemic respiratory failure. However, in other settings of acute hypoxemic respiratory failure, such as acute lung injury and acute respiratory distress syndrome, this has not been shown. The use of IPPV may improve outcomes in patients with severe cardiogenic shock. However, IPPV has not proven to be beneficial in traumatic brain injury and flail chest, in the absence of other indications. PMID- 11874606 TI - Setting the frequency-tidal volume pattern. AB - Alveolar (and thus arterial) P(O2) and P(CO2) clearly depend on minute ventilation. However, we need to balance gas exchange goals against the risk of overstretching, especially of the healthier regions of the lung. The plateau pressure is probably the best easily-obtained marker of the risk of stretch in the lung, and a commonly quoted threshold is 30--35 cm H(2)O, the normal maximum transalveolar pressure at total lung capacity. In establishing the proper balance of stretch versus gas exchange, we need to address what levels of pH and P(aO2) we consider acceptable. There are no good data to guide us on the lowest tolerable pH, but 7.2 is commonly quoted in the literature, and 7.15 was the lower limit of acceptability in the ARDS (acute respiratory distress syndrome) Network trial. P(O2) levels as low as 55 mm Hg may be well tolerated, provided there is reasonable oxygen delivery. In distributing the desired minute volume between respiratory frequency and tidal volume (V(T)), a V(T) of 6 mL/kg ideal body weight has been shown to improve ARDS outcome, compared to 12 mL/kg. Thus, 6 mL/kg should be the "start point." Adjustments upward could be considered the plateau pressure is acceptable, in order to improve gas exchange or comfort. Conversely, downward adjustments should be considered if the plateau pressure is high and the gas exchange is acceptable. Frequency is adjusted for the desired minute ventilation. It must be recognized, however, that as frequency (and minute ventilation) increases, the risk of air trapping and intrinsic positive end expiratory pressure (PEEP) increases. Just like applied PEEP, intrinsic PEEP increases the baseline pressure and stretch upon which the V(T) is delivered. The end-inspiratory stretch increases accordingly. The shape and duration of the flow pattern may affect gas mixing, recruitment, cardiac function, intrinsic PEEP buildup, and patient comfort. It is also conceivable that certain flow patterns can produce an acceleration injury. Although small clinical trials using physiologic end points espouse certain flow patterns, there are no good outcome data at present supporting any particular approach. Some authors suggest that high-frequency ventilation (HFV) might be considered an "ultimate" lung protective strategy. HFV creates considerable intrinsic PEEP, which, when coupled with sustained inflation maneuvers, can provide substantial alveolar recruitment. In addition, the small V(T) of HFV prevents excessive end-inspiratory distention. Although considerable clinical data support the use of HFV in pediatric patients at risk for ventilator-induced lung injury, there are few data from adults. Whether HFV will prove valuable in well-designed open lung strategies in the adult population still has to be determined. PMID- 11874607 TI - How to set positive end-expiratory pressure. AB - Application of positive end-expiratory pressure (PEEP) in acute lung injury patients under mechanical ventilation improves oxygenation and increases lung volume. The effect of PEEP is to recruit lung tissue in patients with diffuse lung edema. This effect is particularly important in patients ventilated with low tidal volumes. Measurement of respiratory system mechanics in patients with acute respiratory distress syndrome is important to assess the status of the disease and to choose appropriate ventilator settings that provide maximum alveolar recruitment while avoiding overdistention. In patients with acute respiratory distress syndrome in whom the lungs have been near-optimally recruited by PEEP and tidal volume, the use of recruitment maneuvers as adjuncts to mechanical ventilation remains controversial. The application of PEEP in patients with unilateral lung disease may be detrimental if PEEP hyperinflates normal lung regions, thus directing blood flow to diseased lung regions. In patients with air flow limitation and lung hyperinflation, the application of additional external PEEP to compensate for intrinsic PEEP and flow limitation frequently decreases the inspiratory effort to initiate an assisted breath, thus decreasing breathing work load. PMID- 11874608 TI - The role of spontaneous breathing during mechanical ventilation. AB - The tremendous progress in microprocessor-driven ventilator technology over the last years has facilitated the introduction of a broad variety of different ventilatory modes into the clinical practice of mechanical ventilation. Many of these newer modalities are designed for partial ventilatory support, which might reflect the complexity of the issue of patient ventilator interactions when spontaneous breathing activity is present compared to controlled mechanical ventilation. There are reasons to believe that allowing some degree of spontaneous breathing activity during mechanical ventilation is useful not only to gradually withdraw ventilatory assistance in the process of weaning but also to avoid some of the adverse effects of mechanical ventilation in the early phase of acute respiratory failure when classically controlled modes of ventilation are used. It is the aim of this article to review the effects of preserved spontaneous breathing activity during mechanical ventilation with different ventilatory modalities in acute respiratory failure patients. PMID- 11874609 TI - Lung recruitment: the role of recruitment maneuvers. AB - There is increasing appreciation that lung-protective strategies are beneficial in patients with acute respiratory distress syndrome. Using low tidal volume in these patients improves survival. However, low tidal volume ventilation may promote alveolar de-recruitment. This has led some to advocate the use of "open lung" strategies that stress the use of high positive end-expiratory pressure levels and recruitment maneuvers. A recruitment maneuver is a sustained increase in airway pressure with the goal to open collapsed lung tissue. A variety of approaches have been used as recruitment maneuvers, including increasing the level of positive end-expiratory pressure, sustained inflation maneuvers, sigh breaths, spontaneous breathing, and others. There have been a number of recent reports describing improvements in arterial oxygenation with the use of recruitment maneuvers. However, the impact of recruitment maneuvers on patient important outcomes such as survival is unknown. PMID- 11874610 TI - Ventilatory adjuncts. AB - A number of adjuncts to mechanical ventilation have been the focus of recent research. Automatic tube compensation (the regulation of airway pressure by estimation of tracheal pressure) appears to be an ideal approach to unloading the resistive effort imposed by the endotracheal tube. Randomized controlled trials have recently been performed with high frequency oscillation (HFO), partial liquid ventilation (PLV), and prone positioning. Unfortunately, all of those trials were negative; however, it appears the only technique that will be abandoned for the near future is PLV. The HFO trial trended toward benefit with HFO, and one must question the protocol used in the prone positioning trial. With both HFO and prone positioning we will have to wait for additional randomized clinical trials before the status of those techniques can be determined. No randomized trials of tracheal gas insufflation have been performed. Of major concern with tracheal gas insufflation is the lack of a commercial product. PMID- 11874611 TI - Sedation and paralysis during mechanical ventilation. AB - Treatment of anxiety and delirium, provision of adequate analgesia, and, when necessary, amnesia in critically ill patients is humane and may reduce the incidence of post-traumatic stress disorders. Injudicious use of sedatives and paralytics to produce a passive and motionless patient, however, may prolong weaning and length of stay in the intensive care unit. This report reviews indications and choices for pharmacologic treatment of anxiety, delirium, agitation, and provision of anesthesia in critically ill patients. The choice of pharmacologic agents is made difficult by complex or poorly understood pharmacokinetics, drug actions, and adverse effects in critically ill patients. Advantages, adverse effects, and limitations of drug treatment, including use of neuromuscular blocking drugs and use of sedatives and analgesia during the withdrawal of life-sustaining measures are reviewed. PMID- 11874612 TI - Remediation of "working attention" in mild traumatic brain injury. AB - Several studies have reported beneficial effects of treatments for attentional deficits following traumatic brain injury. Improvements in speed of processing appear to be less robust than improvements on non-speeded tasks, while several studies suggest greater benefits of training more complex forms of attention. The present study presents preliminary results concerning the effectiveness of an intervention for attentional deficits after mild traumatic brain injury. The treatment was based upon the conceptualization of deficits and interventions as a function of the central executive component of working memory, or "working attention" . A prospective, case-comparison design was employed comparing four treatment participants with an untreated comparison sample. Treatment tasks were derived from experimental procedures which have been demonstrated to elicit working memory demands, consisting of "n-back", random generation, and dual-task procedures. The intervention emphasized the conscious and deliberate use of strategies to effectively allocate attentional resources and manage the rate of information during task performance. Treatment participants were more likely to exhibit clinically significant improvement on measures of attention and reduction of self-reported attentional difficulties in their daily functioning. Further analysis suggested that the principal effect of the intervention was on working memory, i.e. the ability to temporarily maintain and manipulate information during task performance, with no direct effect on processing speed. The results are consistent with a strategy training model of remediation, in which the benefits of treatment are due to participants' improved ability to compensate for residual deficits and adopt strategies for the more effective allocation of their remaining attentional resources. PMID- 11874613 TI - Insomnia in a post-acute brain injury sample. AB - OBJECTIVE: The purpose was to establish the frequency of insomnia within the post acute TBI population and compare it with insomnia rates among other rehabilitation outpatients. DESIGN: A prospective study was undertaken of 50 consecutive post-acute TBI admissions and a comparison group of 50 rehabilitation outpatients evenly divided between spinal cord injury (SCI) and musculoskeletal (MSK) cases. SETTING: Subjects were recruited at various outpatient clinics of a major rehabilitation hospital. PATIENTS: Among the TBI subjects, the predominant cause of injury was motor vehicle accident; both mild and severe injuries were well represented in the sample; and, on average, patients were almost 4 months post-injury. The comparison and TBI groups did not differ significantly with respect to education or marital status. However, the MSK group was older and a higher proportion of the SCI group was female. MEASURES: The Pittsburgh Sleep Quality Index (PSQI), Beck Depression Inventory (BDI), and sleep diaries were administered to the TBI group. Only the PSQI and BDI were completed by the comparison group. RESULTS: Thirty per cent of the patients were found to suffer from insomnia. Sleep initiation was a problem almost twice as often as sleep duration. An additional 12% did not meet the DSM-IV criteria for insomnia but, nevertheless, experienced a degradation of sleep quality, as measured by the PSQI. Conversely, only slightly more than half (58%) of the TBI sample reported sleep to be relatively normal and satisfactory. Insomnia was also commonly reported by the patients in the rehabilitation comparison groups. They generated significantly higher mean PSQI Global Scores relative to the TBI group and the frequency of poor sleep quality was elevated significantly above the TBI rate. Relative to the TBI cases, twice as many comparison group patients were classified by the PSQI as insomniacs. CONCLUSION: Poor sleep quality and insomnia were definitely problems for the TBI group, although the magnitude of these problems was much greater for the rehabilitation comparison group. Degraded and disordered sleep may represent widespread challenges within the rehabilitation population in general. PMID- 11874614 TI - Functional recovery and instrumental activities of daily living: follow-up 1-year after treatment in a stroke unit. AB - The objective of the study was to assess the utility of the Frenchay Activities Index (FAI) to measure instrumental activities of daily living (IADL) and functional recovery in stroke patients compared to other measures such as Barthel Index (BI) and Scandinavian Stroke Scale (SSS). A cross-sectional descriptive analysis design was done. Ninety stroke survivors (41 women, 49 men; mean age 68 years) discharged from the Stroke Unit at San Carlos Hospital, Madrid, were assessed by BI at discharge and by BI and FAI 1-year after stroke. At discharge, 40% had total or severe disability (BI < or =60) and at 1-year 11.1%. FAI (mean value 36 +/ 11) correlated with Barthel index, capacity for walking, strength in upper limb and total SSS 1-year after stroke (p < 0.0001). Fifty-two per cent of stroke patients became independent in their ADL during the first year. BI was the strongest predictor of independence in FAI Social activities-category. PMID- 11874615 TI - The vegetative and minimally conscious states in children: spasticity, muscle contracture and issues for physiotherapy treatment. AB - The neuropathology of the vegetative (VS) and minimally conscious (MCS) states and the pathophysiology of spasticity are reviewed. Current treatment options available in the physical management of children in a low-level state and factors influencing the physiotherapy treatment of children in a low-level state will be discussed. The complex neuropathology of VS and MCS helps to explain the varied clinical presentations of children in VS and MCS. Spasticity and muscle contracture are common motor sequelae of VS and MCS. Loss of inhibition by descending motor pathways is thought to result in increased muscle tone or spasticity. However, secondary changes in muscle fibre structure and periarticular connective tissue may be an additional component to increased muscle tone. A multimodal approach combining physical, pharmacological and surgical interventions is likely to be the most effective. Knowledge of the likelihood of recovery from VS and MCS can be helpful in determining the frequency and intensity of physiotherapy. Ethical issues in the management of children in a low-level state include a consideration of the benefits to the child and the child's family and the costs to the health care team and the medical institution. PMID- 11874616 TI - Altered electrodermal response to facial expression after closed head injury. AB - The objective was to determine whether diffuse damage to orbital and ventromedial regions of the prefrontal cortex usually associated with moderate-to-severe closed head injury (CHI) would affect the ability to perceive and respond to socially relevant information. METHODS: Participants with CHI and age-matched non injured controls were presented with faces that varied with respect to emotional expression while electrodermal activity (EDA) was monitored. Cognitive and general adaptive functioning was also assessed. RESULTS: CHI was associated with a failure to increase EDA in response to negative facial expressions and with reduced ability to identify negative expressions, especially fear. The groups differed on other signs of orbital/medial prefrontal damage such as anosmia and in general social adaptability and awareness-of-deficit as measured by the Brock Adaptive Functioning Questionnaire. CONCLUSIONS: CHI affected the ability to identify and respond to negative facial expression. Addressing these deficits may enhance rehabilitative efforts within the social domain. PMID- 11874617 TI - Partial weight-bearing gait retraining for persons following traumatic brain injury: preliminary report and proposed assessment scale. AB - The objectives of this investigation were to (1) document the recovery patterns of walking ability in two patients recovering from traumatic brain injury receiving partial weight-bearing gait retraining, and (2) introduce a new assessment scale of gait progress for patients receiving partial weight support therapy. The two patients were categorized as acute (< 6 months) and chronic (> 2 years) injury. Each patient received extensive in-patient rehabilitation, including physical therapy designed with twice-weekly partial body support gait training. The subjects made improvements in all measured indicators of gait ability (i.e. muscle strength, spasticity, standing balance). However, assessment of their improvement using standard assessment scales showed little progress. The newly devised Missouri Assisted Gait (MAG) scale, which includes developmental components of gait ability measured dramatic gains. This added precision of measurement was useful in communicating progress to both patients and providers. PMID- 11874618 TI - Agitation assessment in severe traumatic brain injury: methodological and clinical issues. AB - The aim of this single case study was to evaluate the applicability of a graphic and statistical time-series analyses in the observation of an agitation disturbance in a 16-year-old patient who had sustained a severe traumatic brain injury. The agitation was measured using the Agitated Behaviour Scale. The experimental model was of the A-B type: phase A corresponded to the period of vegetative state, and phase B to the period following the reawakening from coma. The data were submitted to visual and statistical analysis by the split-middle trend line method, function of autocorrelation, and C statistic. The results show the different nature and frequency of the agitated behaviour during the vegetative state and after reawakening from coma. The application of a statistical analysis to establish whether the behavioural disturbance is random or a response to the environment allows the adoption of specific and potentially more efficacious treatments. PMID- 11874619 TI - Does inbreeding lead to decreased human fertility? AB - In most Western countries there is a widespread belief, fostered in part by historical prejudice and religious proscription, that inbreeding in human populations causes a reduction in fertility. Support for this belief has been claimed in HLA-based studies, with increased rates of fetal losses suggested in HLA-compatible unions. To critically assess the overall status of fertility in consanguineous unions, data on 30 populations resident in six countries were collated from a systematic review of the literature. The mean numbers of live births were then compared in four consanguinity test categories, ranging from second cousin to uncle-niece/double first cousin, and corresponding non consanguineous reference groups. Linear regressions indicated a positive association between consanguinity and fertility at all levels of inbreeding, attaining statistical significance at first cousin level (p < 0.0001). The results were, however, subject to a number of potential limitations, in particular lack of control for important socio-demographic variables. To overcome this problem, data on first cousin marriages were abstracted from the National Family and Health Survey conducted in India during 1992-1993. Multivariate analysis showed that fertility in first cousin unions was positively influenced by a number of variables, including illiteracy, earlier age at marriage and lower contraceptive uptake, but the most important of these parameters were duration of marriage and reproductive compensation. In net terms, consanguinity was not found to be associated either with a significant positive or negative effect on fertility. PMID- 11874620 TI - Prediction equations for handgrip strength in healthy Indian male and female subjects encompassing a wide age range. AB - PRIMARY OBJECTIVE: Handgrip strength is a simple index of skeletal muscle function and a functional index of nutritional status. A major lacuna in the use of handgrip strength is the limited availability of normative data. The main objective of this paper was to develop prediction equations for handgrip strength in Indians covering a wide age range. METHODS: Handgrip strength and basic anthropometric parameters were measured in 1024 healthy Indian subjects of both genders (613 males, 411 females) between the ages of 5 and 67 years. The sample was randomly divided into two sets; one set (n = 677) was used to develop the prediction equations for handgrip strength and the other (n = 347) was used to validate the equations. Each data set was further divided into two subsets (adults > 18 years, sub-adults 30 mm Hg, 1 mm Hg=0.133 kPa). The CPP also had a prognostic power on neurological deterioration when its level less than 60 mm Hg. CONCLUSIONS: It suggests that it's very important to lower the intracranial hypertension and keep the CPP not less than 60 mm Hg during the t reatment of STBI. PMID- 11874684 TI - Effect of endothelin-1 on hepatic damage induced by endotoxin. AB - OBJECTIVE: To observe effect of endothelin-1 (ET-1) on hepatic damage induced by endotoxin. METHODS: A total of 90 rats were randomly divided into control group (group C), endotoxin treated group (group LPS) and endotoxin plus ET-1 antibody treated group (group LEA). An observation was done on the changes of ET-1 concentration, and transcription and expression of ET-1 mRNA. Plasma glutamic pyruvic-transaminase enzyme (GPT), hepatic lactate dehydrogenase (LDH), adenosine triphosphate (ATP) and malondialdehyde (MDA) were also observed at 3, 6, 9, 12, 24 hours after saline, endotoxin (10 mg/kg) and ET-1 antibody (dalubine 1:2000, 2 ml/kg) administration. RESULTS: The results indicated that the concentration of plasma and hepatic ET-1 and expression of ET-1 mRNA in liver significantly increased following endotoxemia. The hepatic ET-1 levels were inversely correlated with the ATP concentration, and positively related to the MDA concentration. ET-1 antibody could partially protect the liver against damage induced by endotoxin. CONCLUSIONS: These results suggest that endotoxin may, on transcription and translation level, lead to an increase of ET-1 in synthesis. ET 1 may contribute to hepatic damage during endotoxemia. PMID- 11874683 TI - Effect of dexamethasone by local treatment on cerebral edema and serum myelin basic protein after brain injury in rabbits. AB - OBJECTIVE: To explore the effect of dexamethason e by local treatment on cerebral edema and brain damage after brain injury. METHODS: Twenty-two rabbits were classified into 2 groups, Gro up A (the control group, n=11) and Group B (the treated gr oup, n=11). An rabbit brain contusion model was made by bo ne windowplasty by extradural hitting. Group B was treated by local infiltrating and spraying of dexamethasone at equidistance to lesions. Group A was given nor mal saline in the same way as Group B. The changes of moisture in brain tissues and serum myelin basic protein (MBP) were observed. RESULTS: The percentage of water content in damaged hemisphere in Group A and Group B was 81.75%plus minus0.56% and 79.45%plus minus0.52% respe ctively. There was a significant difference between the 2 groups (P<0.05). The normal level of MBP was 1.66 mug/Lplus minus0.71 mug/L, while the value of MBP in Group A and Group B were 5.98 mug/Lplus minus2.08 mug /L and 3.15 mug/Lplus minus1.09 mug/L separately. The level of MBP in Group A an d Group B were higher than normal level and there was also a significant differe nce between Group A and Group B (P<0.05). CONCLUSIONS: The results of our study showed that the brain moi sture and MBP in serum were increased after brain injury while reduced after tre atment with dexamethasone. It is demonstrated that local treatment of brain inju ry with dexamethasone has an obvious therapeutic effect on cerebral edema and se rum MBP. PMID- 11874685 TI - Simulation study on the effectiveness of accident proneness test in drivers. AB - OBJECTIVE: To explore the effectiveness of accident proneness test in drivers. METHODS: Accident proneness test results in 100000 drivers were simulated by function of RANNOR using SAS statistical software, their accident records were simulated by function of UNIFORM according to 3 alternatives, the accident rate of qualified drivers was 0.01, and the accident rate of unqualified drivers was 0.01, 0.05 and 0.10, respectively. RESULTS: It was found that there was no effectiveness of accident proneness test if the accident rate of unqualified drivers was equal to that of the qualified ones, if the accident rate of the unqualified drivers was really higher than that of the qualified ones, the test could identify a certain proportion of high risk drivers. CONCLUSIONS: Accident proneness test may be effective to some extent, it is advisable to carry out further prospective study or pilot work so as to practically prove the effectiveness of the test. PMID- 11874686 TI - Experimental study on hemorheological and pathological changes following severe myocardial contusion. AB - OBJECTIVE: To investigate the mechanism of severe myocardial contusion in rabbits. METHODS: A total of 32 New Zealand rabbits were randomly divided into 2 groups, the severe myocardial contusion group (the experimental group, n=16) and the sham-impact control group (the control group, n=16). Hemorheological parameters, interleukin-8 (IL-8) in serum, the water contents of myocardium and polymorphonuclear neutrophil (PMN) infiltration in contused myocardium were observed at 24 hours after the experiment. RESULTS: As compared with the control group, the hemorheological parameters in the experimental group including the whole blood viscosity (etab), erythrocyte aggregation index (EAI), hematocrit (HCT), serum fibrinogen (Fib), Casson viscosity (Gammay) and erythrocyte sedimentation rate (ESR), significantly increased. The IL-8, PMN infiltration and the water contents of the contused myocardium also significantly increased. CONCLUSIONS: It suggests that the hemorheological disorder, increase of IL-8 in serum, and PMN infiltration in contused myocardium may contribute to the development of cardiac edema and secondary myocardial damage following severe myocardial contusion in rabbits. PMID- 11874687 TI - A new approach to tissue repair: gene therapy. AB - The process of tissue repair involves a complex tissue response to injury in which growth factors, playing a major role in this process, trigger, control and terminate soakage of inflammatory cells, cells proliferation, secretion of matrix and scars formation by autocrine, paracrine or both. Thus, growth factors can be used to alter the microenvironment of the wounded tissues and to promote their repair. But, there are notable disadvantages in using purified recombination growth factors, 1) the source is so limited that their prices are expensive; 2) the ir half-lives are short and easy to be destroyed by wound proteases; 3) there is no perfect carrier; 4) high initial doses are required but easy to bring toxicity; 5) it is difficult to apply growth factors in deep wounded tissues again and again, their function cannot be played enough accordingly; 6) most of growth factors are the products of recombination. All above-mentioned disadvantages result in a low activity. PMID- 11874688 TI - Traumatic basilar-cavernous fistula associated with aneurysm of basilar artery. AB - A 19-year-old boy's left temporal region was struck by a screwdriver, he immedietly lost consciousness for several minutes, when he came back he had a serious headache and obvious left ptosis. CT scanning showed an intracranial air accumulation and obvious traumatic subarachnoid hemorrhage (SAH), 2 weeks later magnetic resonance imaging (MRI) and magnetic resonance angiography (MRA) showed a traumatic aneurysm in basilar artery. Digital subtraction angiography (DSA) was performed 4 weeks later, revealing a basilar-cavernous fistula and a saccular aneurysm of the basilar artery in his head. After the patient was treated with endovascular embolization therapies twice and with mechanical detachable spiral (MDS) for 5 months the patient was cured finally. PMID- 11874689 TI - Identification of the adrenal protease that cleaves pro-gamma-MSH: the dawning of a new era in adrenal physiology? AB - In respect to growth, the adrenal is a dynamic organ that requires constant stimuli from pituitary-derived POMC peptides to maintain its tonic state since either hypophysectomy or dexamethasone treatment results in rapid adrenal atrophy. It has been previously demonstrated that peptides derived from the N terminus of the 16 kDa fragment of POMC not containing the gamma-MSH sequence are potent adrenal mitogens both in vitro and in vivo. However, since these shorter peptides are not found in the circulation, it has been suggested that they are generated by cleavage of the 16 kDa fragment by a specific protease expressed by the adrenal. This putative enzyme has recently been identified and this commentary describes the findings to date and highlights some of their possible implications. PMID- 11874691 TI - IGF-binding protein-5: flexible player in the IGF system and effector on its own. AB - The multiple activities of IGF-I and -II are modulated by a family of IGF-binding proteins (IGFBP-1 to -6). Although structurally related, each IGFBP has unique properties and exerts specific functions. IGFBP-5 is the most conserved IGFBP across species and was identified as an essential regulator of physiological processes in bone, kidney and mammary gland. In addition, IGFBP-5 appears to play a decisive role in the control of proliferation of specific tumour cell types. In many situations IGFBP5 exerts biological activities in the absence of IGFs, indicating the existence of IGF-independent actions. This concept was supported by the unexpected localisation of IGFBP-5 in the nucleus and the description of IGFBP-5-specific membrane-bound IGFBP-5 receptor(s). The scope of this review is to summarise the available information about the structure of IGFBP-5 and the regulation of its expression. Furthermore, the potential significance of IGFBP-5 in the regulation of physiological processes will be critically analysed in the light of recent experimental data. PMID- 11874692 TI - Comparative effects of testosterone propionate, oestradiol benzoate, ICI 182,780, tamoxifen and raloxifene on hypothalamic differentiation in the female rat. AB - Hypothalamic differentiation in the female rat during the neonatal period is critically dependent on the steroid milieu, as permanent changes in reproductive function are observed after administration of oestradiol and testosterone during such a critical stage. Selective oestrogen modulators (SERMs) constitute a family of drugs that, depending on the tissue, are able to exert oestrogenic or antioestrogenic actions. The present experiments were conducted to analyse whether the SERMs, tamoxifen and raloxifene, can cause oestrogenic actions during the hypothalamic differentiation period. Postnatal female rats were injected between days 1 and 5 with 100 microg/day tamoxifen, raloxifene or ICI 182,780 (a pure antioestrogen). Other groups of animals were injected on day 1 of age with 100 microg oestradiol benzoate (OeB) or 1.25 mg testosterone propionate (TP) alone or in combination with raloxifene (500 microg/day between days 1 and 5). In all experimental groups, the age, body weight and concentrations of serum gonadotrophins at vaginal opening were recorded, whereas vaginal cyclicity and the negative and positive feedback between oestradiol and LH were monitored in adulthood. The results obtained confirmed the ability of high doses of OeB or TP to alter the normal differentiation of the brain permanently. They also reinforced the hypothesis that oestrogens are also necessary for normal brain differentiation in female rats because administration of a pure antioestrogen, such as ICI 182,780 permanently altered the function of the reproductive axis. In addition, our data provided evidence for different actions of the two SERMs under analysis (raloxifene and tamoxifen) upon peripheral targets, as raloxifene advanced vaginal opening whereas tamoxifen did not. In contrast, their actions on brain differentiation appeared similar and analogous to those obtained after neonatal administration of oestradiol, as evidenced by vaginal acyclicity, ovarian atrophy, sterility and abolition of negative and positive feedback between oestradiol and LH, thus suggesting an oestrogenic action of these SERMs on hypothalamic differentiation. Moreover, the oestrogenic activity of raloxifene was supported by its inability to block the effects of OeB and TP administered neonatally. In conclusion, the present results indicated that the SERMs, tamoxifen and raloxifene, exert an oestrogen-like effect upon hypothalamic differentiation of the neonatal female rat. PMID- 11874690 TI - Pro-opiomelanocortin processing in the hypothalamus: impact on melanocortin signalling and obesity. AB - Bioactive peptides derived from the prohormone, pro-opiomelanocortin (POMC), are generated in neurons of the hypothalamus and act as endogenous ligands for the melanocortin-4 receptor (MC4R), a key molecule underlying appetite control and energy homeostasis. It is therefore important to understand many aspects of POMC gene regulation in the brain, as pharmacological manipulation of POMC expression/processing could be a potential strategy to combat obesity. Most studies that have analysed POMC gene expression in the hypothalamus have focused on gene transcription experiments. Ultimately, however, factors that regulate post-translational processing and secretion of peptides will have most bearing on melanocortin signalling. This article focuses on (a) current evidence that POMC is involved in obesity, (b) how POMC transcription is regulated in the hypothalamus, (c) the mechanism by which proteolytic processing of POMC is controlled in the hypothalamus and what peptides are produced and (d) which POMC derived peptides are the most potent ligands at the melanocortin receptor in vitro and in vivo. It seems that post-translational cleavage of POMC in the hypothalamus may be regulated with respect to energy requirement. We predict that further research into hypothalamic POMC processing, and the proteolytic enzymes involved, may yield important new clues on how flux through the MC4R pathway is regulated. PMID- 11874693 TI - Glucocorticoids are not necessary for the inhibitory effect of endotoxic shock on serum IGF-I and hepatic IGF-I mRNA. AB - The aim of this work was to elucidate the possible role of glucocorticoids in the bacterial lipopolysaccharide (LPS)-induced decrease in hepatic IGF-I synthesis. For this purpose, we studied the effect of LPS on IGF-I in two rat strains, Wistar and Lewis, which have different adrenal responses to inflammation. Compared with Wistar rats, Lewis rats have a reduced hypothalamic-pituitary adrenal response to inflammatory stimuli. Rats received two i.p. injections of 1 mg/kg LPS and were killed 4 h after the second injection. LPS induced an increase in serum concentrations of both ACTH and corticosterone, the increase being more pronounced in Wistar than in Lewis rats. LPS decreased hepatic GH receptor (GHR) and IGF-I mRNA only in Wistar rats. However, serum concentrations of IGF-I were significantly decreased (P<0.01) in both Wistar and Lewis rats. These data indicate that the adrenal axis may mediate the inhibitory effect of LPS on GHR and IGF-I synthesis in the liver. In a second experiment, adrenalectomized or sham-operated Wistar rats were injected with LPS. Two LPS injections (0.1 mg/kg) decreased serum concentrations of IGF-I in both type of rat; however, the inhibitory effect of LPS on liver GHR and IGF-I mRNA was observed in adrenalectomized rats, but not in intact rats. All these data suggest that some component of the adrenal axis, other than glucocorticoids, mediates the inhibitory effect of LPS on liver GHR and IGF-I. PMID- 11874694 TI - GH is a regulator of IGF2 promoter-specific transcription in human liver. AB - The regulation of the insulin-like growth factor-II gene (IGF2) is complex and involves the usage of four promoters resulting in different 5' untranslated regions, but with a common translated product. The IGF2 gene product is a mitogenic and survival factor that has been suggested to be important for a normal fetal development and cancer. In this paper we present evidence suggesting that the human IGF2 gene is regulated by GH, and that this regulation occurs in a promoter-specific way. Three lines of evidence support this finding. First, in vivo data from patients treated with GH (one injection or daily injections for 5 consecutive days) showed an increase in the IGF2 P2 promoter derived transcript after acute treatment, and of the P4 promoter transcript after short-term treatment while the P1 promoter derived transcript did not show any significant change. Secondly, isolated human liver cells treated with GH for 2 h displayed an upregulation of the P2 promoter derived transcript. Thirdly, employing transfection experiments in GH-receptor positive CHO cells with P2 and P4 promoter-luciferase constructs, an upregulation by GH was evident, while a P1 promoter construct was unresponsive. We suggest that GH may be a physiological regulator of IGF2 in humans. PMID- 11874695 TI - The IGF system in neuroblastoma xenografts: focus on IGF-binding protein-6. AB - With a view to investigating the implication of IGF-binding protein-6 (IGFBP-6) in the growth of neuroblastomas, nude mice were injected with IGFBP-6-expressing or control IGR-N-91 human neuroblastoma cells and the resulting xenografts examined. Expression of IGFBP-3, IGFBP-4 and type 1 and type 2 IGF receptor messengers was similar in control tumours and equal-sized IGFBP-6-expressing tumours that had developed. IGF-II was more strongly expressed in control tumours, and IGFBP-6-expressing tumours contained less IGFBP-2 than controls. In both populations, there was a significant positive correlation between IGF-II and IGFBP-2 expression. In small IGFBP-6-expressing xenografts where tumour development had apparently been arrested, haematoxylin--eosin and TUNEL staining revealed numerous apoptotic cells. In situ hybridization indicated homogeneous distribution of the IGFBP-6 signal in test tumours. In cell culture, IGFBP-6 expressing cells expressed similar amounts of IGFBP-2, IGF-II and N-myc mRNAs as control cells; but media conditioned by IGFBP-6-expressing cells contained less intact IGFBP-2 protein, with no increase in its proteolytic fragment. In media treated with plasminogen, in which IGFBP-2 was proteolysed, IGFBP-6 was increased. With its especially strong affinity for IGF-II and its resistance to proteolysis, IGFBP-6 would act by sequestering IGF-II, hence inhibiting its mitogenic and anti-apoptotic effects. In excess, IGFBP-6 would displace IGF-II from IGFBP-2 whose potentiation of IGF-II action would cease and whose susceptibility to degradation would be increased. This study therefore shows that IGFBP-6 plays a role in neuroblastoma cell growth in vivo and in vitro and that stable overexpression of IGFBP-6 leads to alteration of the initial balance between the IGFBPs. PMID- 11874696 TI - Exogenous expression of Pit-1 in AtT-20 corticotropic cells induces endogenous growth hormone gene transcription. AB - The pituitary-specific POU-homeodomain transcription factor, Pit-1, is known to regulate the expression of the GH gene in somatotropes, prolactin (PRL) in lactotropes, and TSH in thyrotropes. It is not normally expressed in corticotropes or gonadotropes. We addressed the question of whether exogenous Pit 1 was sufficient to induce ectopic transcription of the GH gene in the corticotropic cell line, AtT-20, or the gonadotropic cell line, alpha T3-1. A fusion gene composed of enhanced green fluorescent protein gene and human Pit-1 cDNA was transfected into AtT-20 and alpha T3-1 cells. The endogenous mouse GH mRNA was induced in three of nine AtT-20 cell lines and one of three alpha T3-1 cell lines containing the fusion gene. A small amount of GH protein was also detected in these cell lines. These data indicate that transfected Pit-1 is capable of inducing transcription of the GH gene in AtT-20 cells and alpha T3-1 cells. These data also suggest that synergistic co-factors might be required to transcribe the GH gene effectively for translation into GH protein. Furthermore, our findings support the hypothesis that the function of anterior pituitary cells is determined by the combinatorial action of specific transcription factors. PMID- 11874697 TI - Analysis of expression of chorionic gonadotrophin transcripts in prostate cancer by quantitative Taqman and a modified molecular beacon RT-PCR. AB - Expression of human chorionic gonadotrophin (hCG) is associated with trophoblastic, testicular and other malignancies such as bladder, pancreatic, cervical, breast and prostate cancer. In the prostate, however, hCG expression, associated with neuroendocrine cells, is also found in normal tissue. Of the six highly homologous genes that all encode the beta-subunit of hCG, the beta 7 gene is reportedly the only gene expressed in several non-transformed tissues. The beta 3, 5 and 8 genes would be variably expressed in malignant tissue and placenta, but not in normal tissue. To assess to what extent this expression difference can also be found in the prostate, we compared the levels of the different hCG beta transcripts in concurrent normal and cancerous prostate tissues obtained from 17 patients. To this end, we developed a Taqman real-time fluorescent RT-PCR assay for hCG beta, and a quantitative assay specific for the beta 3, 5 and 8 genes, modified from the molecular beacon principle. This latter assay proved highly specific and capable of reliably distinguishing between these hCG beta transcripts that differ in only one nucleotide. Surprisingly, median expression levels of hCG beta were lower in prostate cancer when compared with normal tissue from the same patient. In contrast, hCG beta 3, 5 and 8 transcripts were found in normal tissue and did not differ in prostate cancer, arguing against a specific role of these transcripts in the development of prostate cancer. PMID- 11874698 TI - Differential expression of human chorionic gonadotropin (hCG) glycosylation isoforms in failing and continuing pregnancies: preliminary characterization of the hyperglycosylated hCG epitope. AB - Human chorionic gonadotropin (hCG) glycoforms change as pregnancy progresses. We have developed an antibody (B152) which can measure a hyperglycosylated early pregnancy isoform of hCG. This putative hyperglycosylated form of hCG arises very early in pregnancies and is rapidly replaced by an isoform that predominates for the remainder of the pregnancy. The profiles of these hCG glycoforms are measured as a ratio of values of two immunometric assays. The profiles of these ratios differ between pregnancies which persist and those which will experience early failure. In this report, daily urine hCG isoform ratios from donor eggs (no exogenous hCG pretreatment), in vitro fertilization pregnancies were profiled and analyzed from the first day following embryo transfer (ET). Significant differences were found between continuing pregnancy and pregnancy loss throughout days 5-20 post-ET. When hCG isoform ratios were analyzed from the first day of detectable hCG, pregnancy loss could be predicted in the case of a single fetus both during the 5- to 10-day time segment (P=0.018) and the 10- to 15-day time segment (P=0.045). When single and multiple fetus pregnancies were analyzed together significance was approached in the 10- to 15-day time period (P=0.058). In a second population of pregnant women who conceived naturally, in whom urine samples were collected at approximately weekly intervals to either term birth or clinical spontaneous abortion, the ratio could discriminate between miscarriages and normal term pregnancies (P=0.043). In later pregnancy, the ratio of hCG isoforms declined more rapidly in miscarriages than in term pregnancy. Antibody B152 was produced using a choriocarcinoma-derived hCG (C5), which was hyperglycosylated at both N- and O-linked sites and was 100% nicked at position beta(47-48). Western blot analyses supported the assay results showing that early pregnancy urine does not contain nicked C5-like hCG. Also, the early pregnancy hCG appeared to be the same size as later pregnancy hCG as judged by SDS gel electrophoresis. A series of Western blot analyses and immunoassays conducted with the samples either non-reduced or reduced showed that B152 is directed to a linear epitope located in the COOH-terminal peptide region of the beta subunit. This indicated that only the O-glycan groups and not the N-linked glycans are part of the antibody epitope. PMID- 11874699 TI - Indomethacin blocks pre-partum nest building behaviour in the pig (Sus scrofa): effects on plasma prostaglandin F metabolite, oxytocin, cortisol and progesterone. AB - In the pig, nest building occurs in the day preceding parturition (gestation=114- 116 days). Nest building behaviour can be induced in pregnant, pseudopregnant and cyclic female pigs following injection of prostaglandin F2alpha. Here we investigated behaviour and endocrine changes after the administration of indomethacin, which inhibits cyclo-oxygenase enzymes and thus prostaglandin synthesis. In experiment 1, pregnant primiparous pigs (gilts) were blood sampled through jugular vein catheters every 20 min from 1000 h on day 113 of pregnancy and behaviour was recorded until birth. Two hours after pre-partum nest building began, animals received 4 mg/kg indomethacin (n=7) or control vehicle (n=8) intramuscularly. Indomethacin-treated animals showed less nest building than controls between 1 and 5 h after injection (P<0.05), during which time they were mostly inactive and lay down for longer than controls. From 5 h before birth until birth there was no significant treatment difference in nest building behaviour. There was a tendency for the start of birth to be delayed in indomethacin-treated animals. Plasma 13,14-dihydro-15-keto-prostaglandin F2 alpha (a major metabolite of prostaglandin F2 alpha) rose during pre-injection nest building and then fell following indomethacin treatment, but was not significantly different between groups when behaviour differed. Plasma oxytocin, cortisol and progesterone were not significantly affected by treatment. In experiment 2, indomethacin-treated non-pregnant gilts (n=7) did not show any changes in activity or posture compared with vehicle-treated controls (n=6) between 90 and 150 min after treatment. These results suggested that indomethacin treatment reversibly and specifically inhibits porcine pre-partum nest building by a mechanism that may involve endogenous prostaglandin F2 alpha synthesis inhibition but is independent of circulating oxytocin, cortisol and progesterone concentrations. PMID- 11874700 TI - Myometrial expression of 11 beta-hydroxysteroid dehydrogenase type 2 in rat pregnancy. AB - The enzyme 11 beta-hydroxysteroid dehydrogenase type 2 (11 beta-HSD2), which reduces glucocorticoid potency in target cells by metabolism of active glucocorticoids, is expressed in the non-pregnant rat uterus in an oestrogen dependent manner. Because glucocorticoids appear to facilitate parturition in many species, expression of 11 beta-HSD2 in pregnant myometrium is likely to influence pregnancy maintenance and possibly the onset and progression of labour. The present study therefore examined myometrial 11 beta-HSD2 mRNA, protein and bioactivity across rat pregnancy, with emphasis on the peripartum period. A single 1.9 kb transcript of 11 beta-HSD2 mRNA was evident in myometrium at all stages, with maximal (P<0.05) levels observed at day 16 (term=day 23). Consistent with this pattern of mRNA expression, Western blot analysis showed the presence of a 40 kDa 11 beta-HSD2 protein at all stages, with the maximal immunoreactive signal also observed on day 16. The 11 beta-HSD2 signal was immunolocalized to myometrial smooth muscle cells and endometrial stromal cells. Bioactivity specific to 11 beta-HSD2 was detectable in myometrium at all stages, but in contrast to the patterns of 11 beta-HSD2 mRNA and protein, the V(max) decreased at the beginning of pregnancy and remained stable until term. The apparent K(m) of 11 beta-HSD2 for corticosterone increased from 47 +/- 11 nM in non-pregnant myometrium to 75 +/- 7 nM by day 10 of pregnancy, and remained high until returning to an intermediate level on the day of delivery (60 +/- 8 nM). Progesterone competitively inhibited 11 beta-HSD2 bioactivity (K(i)=1.75 muM) whereas 20 alpha-hydroxypregn-4-en-3-one, the other major progestin present during rat pregnancy, had no such effect. In conclusion, these data suggest that local levels of active glucocorticoid in the myometrium are determined by the net effects of myometrial 11 beta-HSD-1 and -2 expression across pregnancy. Because the previously reported increase in myometrial 11 beta-HSD-1 near term occurs with little change in myometrial 11 beta-HSD2 bioactivity, this is likely to facilitate parturition by increasing local concentrations of active glucocorticoid. PMID- 11874701 TI - Regulation of 11 beta-hydroxysteroid dehydrogenase type 2 activity in ovine placenta by fetal cortisol. AB - The effect of fetal cortisol on the activity of the type 2 isoform of the enzyme, 11 beta-hydroxysteroid dehydrogenase (11 beta-HSD2), was examined in ovine placenta and fetal kidney by measuring tissue 11 beta-HSD2 activity during late gestation when endogenous fetal cortisol levels rise and after exogenous cortisol administration to immature fetuses before the prepartum cortisol surge. Placental 11 beta-HSD2 activity decreased between 128-132 days and term (approximately 145 days of gestation) in association with the normal prepartum increase in fetal plasma cortisol. Raising fetal cortisol levels to prepartum values in the immature fetus at 128--132 days of gestation reduced placental 11 beta-HSD2 activity to term values. In contrast, 11 beta-HSD2 activity in the fetal renal cortex was unaffected by gestational age or cortisol infusion. When all the data were combined, there was an inverse correlation between the log fetal plasma cortisol level at delivery and placental 11 beta-HSD2 activity, expressed both on a weight-specific basis and per mg placental protein. Fetal cortisol therefore appears to be a physiological regulator of placental, but not renal, 11 beta-HSD2 activity in fetal sheep during late gestation. These findings have important implications, not only for glucocorticoid exposure in utero, but also for the local actions of cortisol within the placental tissues that are involved in initiating parturition in the sheep. PMID- 11874702 TI - Ovariectomy improves neovascularization and microcirculation of freely transplanted ovarian follicles. AB - To investigate the influence of ovariectomy on the physiological process of neovascularization of ovarian follicles, we harvested follicles with a theca interna at 48 h after pregnant mare's serum gonadotropin (PMSG) treatment of Syrian golden hamsters, for subsequent transplantation onto striated muscle tissue of chronically implanted skinfold chambers of non-ovariectomized PMSG synchronized hamsters and bilaterally ovariectomized hamsters. Non-ovariectomized non-PMSG-treated animals served as controls. During a 2 week period after transplantation, neovascularization of these freely transplanted grafts was quantified in vivo by assessment of the newly developed microvascular follicular network, its microvessel density, the diameter of microvessels and their volumetric blood flow using fluorescence microscopic techniques. At day 3 after transplantation, capillary sprouts could be observed in all groups studied, finally developing a complete glomerulum-like microvascular network within 5-10 days. In ovariectomized animals, however, vascularization of follicular tissue was found to be accelerated and enhanced when compared with the follicles in both groups of non-ovariectomized animals. This was associated with significantly higher capillary blood perfusion, which may in part represent the graft's adaptive response to pro-angiogenic stimuli due to elevated gonadotropin levels, but might in particular be mediated by gonadotropin-induced release of vasoactive substances. Interestingly, small preantral follicles lacking a theca interna failed to vascularize in either of the groups. In conclusion, the study demonstrates that ovariectomy improves neovascularization and microcirculation of freely transplanted ovarian follicles. PMID- 11874703 TI - Truncated equine LH beta and asparagine(56)-deglycosylated equine LH alpha combine to produce a potent FSH antagonist. AB - Hybrid hormone preparations were prepared by combining intact and Asn(56) deglycosylated (N(56)dg) equine (e) LH or FSH alpha subunit preparations with truncated, des(121-149)eLH beta (eLH beta t), immunopurified, intact eLH beta or equine chorionic gonadotropin beta (eCG beta) preparations, and eFSH beta. The LH receptor-binding potencies of N(56)dg-eLH alpha:eLH beta t and N(56)dg-eFSH alpha:eLH beta t hybrids were equivalent to that of eLH; however, both N(56)dg alpha preparations were only 3-4% as active as eLH in the rat testis Leydig cell bioassay. In the granulosa cell FSH bioassay, eLH alpha:eLH beta t stimulated progesterone synthesis and induced aromatase activity, while N(56)dg-eLH alpha:eLH beta t was completely inactive at doses up to 5 microg. N(56)dg-eLH alpha:eLH beta t inhibited progesterone production and aromatase induction elicited by 0.3 ng eFSH or 2 ng human (h) FSH. The inhibitory activities of N(56)dg-eLH alpha:eLH beta and N(56)dg-eCG alpha:eLH beta t were only 10% that of N(56)dg-eLH alpha:eLH beta t. N(56)dg-eLH alpha:eCG beta did not inhibit progesterone synthesis stimulated by eFSH at all and appeared to further stimulate aromatase induction at the highest dose tested. Preincubation of N(56)dg-eLH alpha:eLH beta and N(56)dg-eLH alpha:eLH beta t for 72 h at 37 C resulted in no loss of FSH receptor-binding activity. Preincubation resulted in 50% loss of receptor-binding activity by the eFSH preparation due to subunit dissociation, while 88% of N(56)dg-eLH alpha:eFSH beta activity was lost following 72 h, 37 C preincubation. While alpha Asn(56) oligosaccharide had no effect on eLH beta hybrid stability, it did contribute to the stability of the eFSH heterodimer. PMID- 11874704 TI - Biphasic regulation of activin A secretion by gonadotropins in cultured human ovarian granulosa-luteal cells leads to decreasing activin:inhibin ratios during continuing gonadotropin stimulation. AB - Pituitary gonadotropins mediate part of their effects on ovarian function via local hormones and growth factors produced by granulosa cells. Activins and inhibins are among these factors, and they have often opposite effects on various components of the reproductive system. The purpose of this study was to investigate the regulation of ovarian activin A secretion using cultured human ovarian granulosa-luteal cells as a model. The granulosa-luteal cells, obtained from women taking part in an in vitro fertilization program, were cultured and treated with FSH, LH, 8-bromo cAMP (8-BrcAMP, a protein kinase A activator) and 12-O-tetradecanoyl phorbol-13-acetate (TPA, a protein kinase C activator). Conditioned cell culture media were analyzed for activin A, inhibin A and progesterone concentrations with specific enzyme immunoassays. FSH and LH (1-100 IU/l) increased activin A secretion with 24 h of treatment (to 132% and 253% of control respectively; P<0.05 for both), but their effects were inhibitory in 48-h treatments (26% and 16% decreases respectively; P<0.05 for both). In the same experiments, FSH and LH increased inhibin A and progesterone secretion after both 24 and 48 h of treatment. 8-BrcAMP (0.1-100 muM) increased activin A in 24- and 48-h experiments (to 206% and 148% of control respectively; P<0.01 for both). Inhibin A and progesterone secretion were stimulated by 8-BrcAMP time- and dose dependently. TPA increased activin A secretion dose-dependently (0.1-100 ng/ml) in both 24- and 48-h experiments. At 100 ng/ml concentration, it increased activin A up to 61-fold and inhibin A up to 16-fold of control in 24-h experiments. We conclude that gonadotropins regulate immunoreactive activin A secretion biphasically in cultured human granulosa-luteal cells: initial stimulation is followed by inhibition. In contrast, gonadotropins increase inhibin A and progesterone secretion continuously. Consequently, continuing gonadotropin stimulation leads to a decreasing activin:inhibin ratio, which may have a significant role in the local fine-tuning of ovarian steroidogenesis. PMID- 11874705 TI - Pachytene spermatocytes in co-culture inhibit rat Sertoli cell synthesis of inhibin beta B-subunit and inhibin B but not the inhibin alpha-subunit. AB - This study investigates the effects of spermatogenic germ cells on inhibin alpha subunit and beta B-subunit expression, and inhibin alpha-subunit and inhibin B production by rat Sertoli cells in vitro. Sertoli cells isolated from 19-day-old rats were cultured for 48 h at 32 degrees C, in the presence or absence of FSH (2.3-2350 mIU/ml), and in the presence of pachytene spermatocytes, round spermatids or cytoplasts of elongated spermatids purified from adult rat testis by elutriation and density gradient separation. Sertoli cell secretion of inhibin alpha-subunit and inhibin B, as measured by immunoassay, was dose-dependently stimulated by FSH (maximal stimulation 13- and 2-fold, respectively). Round spermatids or cytoplasts co-cultured with Sertoli cells had no effect on basal or FSH-induced secretion of inhibin alpha-subunit or inhibin B. When Sertoli cells were co-cultured with pachytene spermatocytes, inhibin alpha-subunit secretion was unaltered, while inhibin B secretion was suppressed in a cell concentration dependent manner to reach a maximal suppression of 45% compared with Sertoli cells alone (P<0.01). A similar suppression in inhibin B was still observed (64% of Sertoli cells alone) when the pachytene spermatocytes were separated from Sertoli cells by a 0.45 microm pore membrane barrier in bicameral chambers. Pachytene spermatocytes also suppressed FSH-induced inhibin B levels in Sertoli cell co-cultures and this suppression was attributed to a decrease in basal inhibin B production rather than a change in FSH responsiveness. Quantitation of Sertoli cell inhibin alpha- and beta B-subunit mRNA by quantitative (real-time) PCR demonstrated that pachytene spermatocytes did not alter Sertoli cell alpha subunit mRNA expression, but significantly (P<0.01) suppressed basal and FSH induced beta B-subunit mRNA expression to a similar degree to that seen with inhibin B protein levels. It is concluded that pachytene spermatocytes in vitro suppress Sertoli cell inhibin B secretion via factor-mediated suppression of inhibin beta B-subunit expression. These findings support the hypothesis that specific germ cell types can influence inhibin B secretion by the testis independent of FSH regulation. PMID- 11874706 TI - Secretory pattern of inhibin A, inhibin B and inhibin pro-alpha C during induced follicular atresia and subsequent follicular development in the golden hamster (Mesocricetus auratus). AB - The changes in plasma concentrations of inhibins A, B and pro-alpha C were determined in the cyclic golden hamster during follicular atresia induced with antiserum against luteinizing hormone releasing hormone (LHRH-AS) at 1100 h on day 4 (day 1=day of ovulation). Follicular status in the ovary was also studied by determining the number of follicles ovulating in response to human chorionic gonadotrophin (hCG) injection. The time-courses of changes in plasma concentrations of inhibins A, B and pro-alpha C were different from each other during induced follicular atresia and subsequent follicular development. Plasma concentrations of inhibin A decreased to 58.6% of initial values by 24 h after LHRH-AS treatment, and then remained relatively low until at least 60 h later. Plasma concentrations of inhibin B decreased to 64.2% of the initial values by 18 h after LHRH-AS treatment and remained at basal values for 36 h, but increased abruptly to greater than initial values at 42 h after the treatment. Plasma concentrations of inhibin pro-alpha C increased at 6 and 12 h, decreased suddenly to 21.9% of the initial values by 24 h after LHRH-AS treatment, and then gradually increased until 60 h after LHRH-AS. The number of follicles responding to hCG decreased gradually between 0 and 30 h after LHRH-AS, when no ovulations were observed, and then gradually increased until 60 h. The changes in follicular ovulatory responses to hCG correlated with the plasma profile of inhibin A throughout the experiment. These results suggest that inhibin A is mainly secreted by large antral follicles. In contrast, during the subsequent follicular development, the plasma concentration of inhibin B increased earlier than that of inhibin A. These results suggest that inhibin B is secreted by small and large antral follicles. Plasma concentrations of inhibin pro-alpha C were high at a time when plasma concentrations of oestradiol-17 beta had already decreased, indicating that inhibin pro-alpha C is secreted not only from healthy follicles but also from early atretic antral follicles. PMID- 11874708 TI - Oxygen-dependence of ACTH-stimulated aldosterone and corticosterone synthesis in the rat adrenal cortex: developmental aspects. AB - The control of ACTH-stimulated steroidogenesis under decreasing levels of O(2) is not fully understood. The purpose of this study was to examine the effects of decreased O(2) in vitro on rat adrenocortical steroid synthesis at different stages of development. Of interest was the evaluation of the effect of low O(2) on steroidogenesis during the stress hyporesponsive period of the neonate. Rats were killed at 7, 14, or 42 days of age, adrenals collected and capsules (zona glomerulosa, ZG) separated from subcapsules (zona fasciculata/reticularis, ZFR). Cells were dispersed and placed into glass vials each gassed with a different level of O(2) (21, 5, 2, 1, or 0% O(2)). The entire steroidogenic pathway was analyzed by measuring ACTH-stimulated cAMP, corticosterone and aldosterone production during a 2 h incubation. In addition, the early (P450 scc) and late (P450c11 beta and P450 aldo) pathway activities were examined in the presence of cyanoketone. The PO(2) for half-maximal activity (P(50)) for aldosterone synthesis in ZG cells from 7- and 42-day-old rats was approximately 28 mmHg and 7 mmHg respectively, indicating that cells from older rats were more resistant to inhibition by low O(2). The P(50) for cAMP production from the ZG was approximately 14 mmHg for both age groups. The P(50) for corticosterone synthesis was approximately 28 mmHg and <7 mmHg in ZFR cells from 7- and 42-day-old cells respectively. The only enzyme activities affected by low O(2) (<35 mmHg) were P450 aldo and P450 scc. Moderate decreases in O(2) (from approximately 150 mmHg) decreased aldosteronogenesis, possibly due to observed decreases in cAMP generation, but not due to decreases in steroidogenic enzyme activity (7-day old). Severe decreases in O(2) presumably inhibited P450 aldo through a direct effect on enzyme activity (both ages). P450 scc activity (including cholesterol transport) also seems to be decreased by very low O(2) (7-day-old). These findings illustrate a novel developmental alteration in O(2)-regulated steroid production, and may have implications for neonatal health and disease. PMID- 11874707 TI - Interdependence of steroidogenesis and shape changes in Y1 adrenocortical cells: studies with inhibitors of phosphoprotein phosphatases. AB - Y1 adrenocortical cells respond to activators of the cyclic AMP-dependent protein kinase (PKA) signalling pathway not only with increases in steroid secretion but also with a characteristic change in cell morphology from flat and adherent to round and loosely attached. This change of shape, which may facilitate cholesterol transport to the mitochondrion, requires tyrosine dephosphorylation of the focal adhesion protein, paxillin, and can be blocked by inhibitors of phosphotyrosine phosphatase (PTP) activity. In a previous study we demonstrated that inhibition of phosphoserine/threonine phosphatase 1 and 2A (PP1/2A) activities caused a similar morphological response to PKA activation whilst opposing the effects on steroid production. We have now investigated the responses to PKA activation and inhibition of PP1/2A and used PTP inhibitors to examine the relationship between the morphological changes and enhanced steroid production. Both forskolin (FSK) and the PP1/2A inhibitor, calyculin A (CA), caused rapid and extensive rounding of Y1 cells. FSK-induced cell rounding was reversible and accompanied by a reduction in the tyrosine phosphorylation of paxillin. Rounding was prevented by the PTP inhibitors pervanadate (PV) and calpeptin (CP) and was associated with the maintained tyrosine phosphorylation of paxillin. In contrast, CA-induced cell rounding was not reversible over a 2-h period and was not affected by the presence of PTP inhibitors, and CA had no effect on the tyrosine phosphorylation of paxillin. Although neither CA nor FSK produced any gross changes in cell viability as judged by Trypan Blue exclusion or mitochondrial activity, CA-treated cells showed a marked reduction in total protein synthesis assessed by (35)S-incorporation. The effects of FSK and the PTP inhibitors on cell rounding were reflected in their effects on steroid production since PV and CP also inhibited FSK-stimulated steroid production. These results suggest that the mechanism through which inhibition of PP1/2A activities induces morphological changes in Y1 cells is fundamentally different from that seen in response to activation of PKA. They are consistent with PKA-induced shape changes in adrenocortical cells being mediated through increased PTP activity and the dephosphorylation of paxillin, and support the view that the morphological and functional responses to PKA activation in steroidogenic cells are intimately linked. PMID- 11874709 TI - Regulation of prolactin receptor expression in ovine skin in relation to circulating prolactin and wool follicle growth status. AB - Seasonal patterns of hair growth are governed, at least in part, by levels of prolactin in circulation, and although receptors for prolactin (PRLR) have been demonstrated in hair follicles, little is known of their regulation in relation to follicular cycles. In this study, a photoperiod-generated increase in prolactin was used to induce a wool follicle cycle during which changes in PRLR expression in sheep skin were determined by ribonuclease protection assay and in situ hybridisation. mRNA for prolactin and both isoforms of PRLR were also detected in skin by reverse transcription and polymerase chain reaction. As circulating prolactin began to rise from low levels, PRLR mRNA in the skin initially fell. These changes immediately preceded the catagen (regressive) phase of the hair cycle. Further increase in prolactin resulted in up-regulation of PRLR during telogen (dormancy), particularly in the epithelial hair germ, to reach a peak during proanagen (reactivation). In anagen (when follicle growth was fully re-established), PRLR mRNA returned to levels similar to those observed before the induced cycle. Hence, this longer term rise and fall of PRLR expression followed that of plasma prolactin concentration with a lag of 12-14 days. PRLR mRNA was most abundant in the dermal papilla, outer root sheath, hair germ, skin glands and epidermis. Location of PRLR in the dermal papilla and outer root sheath indicates action of prolactin on the growth-controlling centres within wool follicles. These cycle-related patterns of PRLR expression suggest dynamic regulation of PRLR by prolactin, thereby modulating hormonal responsiveness of seasonally growing hair follicles. PMID- 11874710 TI - Genetic engineering of the glucocorticoid receptor by fusion with the herpes viral protein VP22 causes selective loss of transactivation. AB - The development of methods for engineering proteins with novel properties opens the way to manipulating intracellular processes in a therapeutically useful way. Glucocorticoids, acting via glucocorticoid receptors (GR), are potent anti inflammatory agents, acting to oppose nuclear factor kappa B (NF kappa B) function. The herpes viral protein, VP22, has been reported to confer intercellular trafficking activity on 'cargo' proteins, potentially facilitating gene therapy with intracellular proteins. VP22GR, resulting from the addition of VP22 to the N terminal of GR, was equipotent with the wild-type GR in opposing NF kappa B p65-driven expression of an NF kappa B reporter gene. Surprisingly, VP22GR was incapable of inducing transactivation of positive glucocorticoid reporter genes (MMTV-luc and TAT3-luc). Furthermore, the VP22GR had powerful dominant negative activity on both endogenous and exogenous GR transactivation. VP22GR was cytoplasmic in quiescent cells, and after hormone addition underwent nuclear translocation to share the same distribution as the GR. The ability of the VP22GR to selectively confer and enhance glucocorticoid-dependent transrepression of NF kappa B may be of use therapeutically in e.g. transplant rejection, inflammatory arthritis or asthma. PMID- 11874711 TI - Mutation analysis of thyroid peroxidase gene in Chinese patients with total iodide organification defect: identification of five novel mutations. AB - Total iodide organification defect (TIOD), where the iodide in the thyroid gland cannot be oxidized and/or bound to the protein, is caused by a defect in the thyroid peroxidase (TPO) gene. Single strand conformation polymorphism analysis was used to screen for mutations in the TPO gene from five unrelated TIOD patients in Taiwan, and five novel mutations were detected. Three of these were frameshift mutations: a single T insertion between nucleotide position 2268 and 2269 (c.2268-2269 insT) in exon 13 and two single C deletions at nucleotide positions 843 (c.843 delC) and 2413 (c.2413 delC) in exon 8 and 14 respectively. The other two were single nucleotide substitutions (c.G1477>A and c.G2386>T) located in exons 9 and 13 respectively. While the former would result in amino acid substitution (Gly493Ser) in the highly conserved region of the TPO polypeptide, the latter would result in either amino acid substitution (Asp796Tyr) or alternative splicing. Of those identified TPO mutations, c.2268 2269 insT was most prevalent and was detected as heterozygous in all but one TIOD patients. All five TIOD patients investigated in this study were compound heterozygous. The method presented in this study could be used for carrier assessment and mutation analysis of newly identified TIOD patients. PMID- 11874712 TI - Inhibitory effects of octreotide on renal and glomerular growth in early experimental diabetes in mice. AB - It was recently discovered that the streptozotocin (STZ)-diabetic mouse model is characterised by GH hypersecretion in contrast to the STZ-diabetic rat, the former thus mimicking the changes in GH in human type 1 diabetes. Inhibition of circulating and renal IGF-I by long-acting somatostatin analogues reduces renal and glomerular growth and urinary albumin excretion in diabetic rats. The aim of the present study was to examine renal and glomerular growth in early experimental diabetes in mice along with changes in the GH/IGF-I axis following treatment with the somatostatin analogue octreotide. Balb/C(a) mice were randomised into non-diabetic controls, placebo-treated and octreotide-treated diabetic (50 microg/day) mice and examined 7 and 14 days after induction of diabetes. There was no effect of octreotide treatment on body weight, glycaemic control or food intake. However, octreotide treatment significantly inhibited renal and glomerular growth by the end of the study period when compared with placebo treatment. In addition, octreotide prevented an increase in kidney IGF-I by day 7. GH hypersecretion was observed in the diabetic groups but octreotide treatment reduced GH levels compared with placebo treatment by day 14. No significant differences in serum or kidney IGF-binding protein-3 levels were observed between placebo- and octreotide-treated diabetic mice. In conclusion, this new diabetic mouse model mimicking human type 1 diabetes is characterised by GH hypersecretion and the somatostatin analogue octreotide is able to prevent renal and glomerular growth, probably mediated through changes in circulating GH and local kidney IGF-I levels. PMID- 11874714 TI - Secretion of bioactive human insulin following plasmid-mediated gene transfer to non-neuroendocrine cell lines, primary cultures and rat skeletal muscle in vivo. AB - The objective of these studies was to evaluate human insulin gene expression following intramuscular plasmid injection in non-diabetic rats as a potential approach to gene therapy for diabetes mellitus avoiding the need for immunosuppression. A wild-type human preproinsulin construct and a mutant construct in which PC2/PC3 sites were engineered to form furin consensus sites were evaluated in in vitro transfections of hepatocyte (HepG2) and myoblast (C2C12/L6) cell lines, primary rat myoblasts, and dermal fibroblasts. In vivo gene transfer by percutaneous plasmid injection of soleus muscle +/- prior notexin-induced myolysis was assessed in rats. In vitro transfection of non neuroendocrine cell lines and primary cultures with wild-type human preproinsulin resulted in secretion of predominantly unprocessed proinsulin. Employing the mutant construct, there was significant processing to mature insulin (HepG2, 95%; C2C12, 75%; L6, 65%; primary myoblasts, 48%; neonatal fibroblasts, 56%; adult fibroblasts, 87%). In rats aged 5 weeks, circulating human (pro)insulin was detected from 1 to 37 days following plasmid injection and the potential of augmenting transfection efficiency by prior notexin injection was demonstrated (wild-type processing, 87%; mutant, 90%). Relative hypoglycaemia was confirmed by HbA1C (saline, 5.5%; wild type, 5.1%; mutant, 5.1% (P<0.05)). Human (pro)insulin levels and processing (wild-type, 8%; mutant, 53%) were lower in rats aged 9 months but relative hypoglycaemia was confirmed by serum glucose at 10 days (saline, 6.4 mmol/l; wild-type, 6.0 mmol/l; mutant, 5.4 mmol/l). In conclusion, prolonged constitutive systemic secretion of bioactive human (pro)insulin has been attained in non-neuroendocrine cells in vitro and in growing and mature rats following intramuscular plasmid injection. PMID- 11874713 TI - Delays in insulin signaling towards glucose disposal in human skeletal muscle. AB - We explored whether the delay that occurs between a rise in plasma insulin and the increase of glucose disposal occurs before, at, or downstream of steps that are believed to be part of the insulin signaling cascade. Skeletal muscle biopsies were obtained from 16 nondiabetic subjects before, and 20 and 180 min after plasma insulin levels had been augmented in euglycemic hyperinsulinemic glucose clamps. Although plasma insulin had reached 98% of its final concentration within 10 min, insulin receptor kinase (IRK) activity, p85 associated with insulin receptor substrate-1 (IRS-1), IRS-1-associated phosphatidylinositol 3-kinase (PI3K) activity, and Thr(308)-protein kinase B (PKB) phosphorylation in the muscle biopsies at 20 min had reached only 60, 48, 34 and 47% respectively of those at 180 min. This suggests a delay before the level of IRK and little or no delay between IRK and PKB activation. The observation that glycogen synthase activity and glucose disposal at 20 min had both only reached 25% of the respective values at 180 min suggests an additional delay downstream of the investigated signaling steps. PMID- 11874715 TI - Gender- and age-related differences in osteoclast formation from circulating precursors. AB - A number of bone diseases characterised by excessive osteolysis (e.g. osteoporosis and Paget's disease) exhibit a marked gender difference in prevalence and are more common in the elderly population. Bone resorption is carried out by osteoclasts, which are formed by fusion of circulating mononuclear precursor cells of haematopoietic origin. In this study, we have determined whether there are gender- and age-related differences in osteoclast formation from circulating precursors. Peripheral blood mononuclear cells (PBMCs) were co cultured with UMR106 osteoblast-like cells in the presence of macrophage-colony stimulating factor (M-CSF) and 1,25 dihydroxyvitamin D(3) (1,25(OH)(2)D(3)) or cultured alone in the presence of sRANKL (soluble receptor activator of nuclear factor kappa B ligand) and M-CSF. As assessed by the formation of tartrate resistant acid phosphatase (TRAP)-positive (TRAP(+)) and vitronectin receptor positive (VNR(+)) multinucleated cells (MNCs), there was no difference in the number of circulating osteoclast precursors in males and females. Lacunar resorption carried out by osteoclasts formed from these precursors was generally increased in males compared with females (P=0.03). An increase in the number of TRAP(+) and VNR(+) MNCs formed from male PBMCs was noted in response to 1,25(OH)(2)D(3) (P<0.005). An increase in lacunar resorption in cultures of PBMCs (10(5) per well) from males was also noted in response to 10(-9) M 1,25(OH)(2)D(3) (P<0.05) and sRANKL (P=0.05), but not M-CSF. The addition of dexamethasone resulted in a marked increase in osteoclast formation and lacunar resorption in both males and females. Post-menopausal females and males of comparable age showed similar levels of osteoclastogenesis. Pre-menopausal women showed similar levels of osteoclastogenesis but less resorption (P=0.01) compared with males of comparable age. These results show that there are specific gender/age-related differences in osteoclast formation and bone resorption and have implications for evaluating osteoclastogenesis in skeletal diseases such as primary osteoporosis and Paget's disease. PMID- 11874716 TI - Modulation of gene expression in human osteoblasts by targeting a distal promoter region of human estrogen receptor-alpha gene. AB - Estrogen receptor (ER) alpha is expressed during osteoblast differentiation; however, both its functional role in bone metabolism and its involvement in osteoporotic pathogenesis caused by estrogen deficiency are not well understood. Loss of ER alpha gene expression could be one of the mechanisms leading to osteoporosis. Therefore, we investigated a possible modulation of ER alpha gene expression in a human osteoblastic cell line and in four primary osteoblast cultures by using a decoy strategy. Double stranded DNA molecules, mimicking a regulatory region of the ER alpha gene promoter (DNA-102) and acting as a 'silencer' in breast cancer cells, were introduced into osteoblasts as 'decoy' cis-elements to bind and functionally inactivate a putative negative transcription factor, and thus to induce ER alpha gene expression. We found that the DNA-102 molecule was able to specifically bind osteoblast nuclear proteins. Before decoy treatment, absence or variable low levels of ER alpha RNAs in the different cultures were detected. When the cells were transfected with the DNA 102 decoy, an increase in expression of ER alpha and osteoblastic markers, such as osteopontin, was observed, indicating a more differentiated osteoblastic phenotype both in the cell line and in primary cultures. These results showed that the DNA-102 sequence competes with endogenous specific negative transcription factors that may be critical for a decrease in or lack of ER alpha gene transcription. Therefore, osteoblastic transfection with the DNA-102 decoy molecule may be considered a tempting model in a putative therapeutic approach for those pathologies, such as osteoporosis, in which the decrease or loss of ER alpha expression plays a critical role in bone function. PMID- 11874717 TI - Expression and action of the growth hormone releasing peptide ghrelin and its receptor in prostate cancer cell lines. AB - This study has examined the expression of two new facets of the growth hormone axis, the growth hormone secretagogue receptor (GHS-R) and its recently identified putative natural ligand ghrelin, in prostate cancer cells. GHS-R 1a and 1b isoforms and ghrelin mRNA expression were detected by RT-PCR in the ALVA 41, LNCaP, DU145 and PC3 prostate cancer cell lines. A normal prostate cDNA library expressed GHS-R1a, but not the 1b isoform or ghrelin. Immunohistochemical staining for the GHS-R 1a isoform and ghrelin was positive in the four cell lines studied. PC3 cells showed increased cell proliferation in vitro in response to ghrelin to levels 33% above untreated controls, implying a potential tumour promoting role for ghrelin in this tissue. This study is the first to demonstrate the co-expression of the GHS-R and ghrelin in prostate cancer cells. It is also the first study to provide evidence that a previously unrecognised autocrine/paracrine pathway involving ghrelin, that is capable of stimulating growth, exists in prostate cancer. PMID- 11874718 TI - Photosynthesis and respiration of black spruce at three organizational scales: shoot, branch and canopy. AB - To gain insight into the function of photosynthesis and respiration as processes operating within a global ecosystem, we measured gas exchange of mature black spruce (Picea mariana (Mill.) B.S.P.) trees at three organizational scales: individual shoots, whole branches and a forest canopy. A biochemical model was fitted to these data, and physiological parameters were extracted. Pronounced seasonal variation in the estimated model parameters was found at all three organizational scales, highlighting the need to make physiological measurements throughout the year. For example, it took over 100 days for physiological activity to increase from zero during the springtime thaw to its yearly maximum. Good agreement was found between parameter values estimated for the different organizational scales, suggesting that, in the case of aerodynamically rough, largely mono-specific forest canopies, physiological parameters can be estimated from eddy covariance flux measurements. The small differences between photosynthetic parameters estimated at the different scales suggest that the overall spatial organization of photosynthetic capacity is nearly optimized for carbon uptake at each scale. PMID- 11874719 TI - Characterization of a Pinus pinaster cDNA encoding an auxin up-regulated putative peroxidase in roots. AB - As part of a study to identify host plant genes regulated by fungal auxin during ectomycorrhiza formation, we differentially screened a cDNA library constructed from roots of auxin-treated Pinus pinaster (Ait.) Sol. seedlings. We identified three cDNAs up-regulated by auxin. Sequence analysis of one of these cDNAs, PpPrx75, revealed the presence of an open reading frame of 216 amino acids with the characteristic consensus sequences of plant peroxidases. The deduced amino acid sequence showed homology with Arabidopsis thaliana (L.) Heynh., Arachis hypogaea L. and Stylosanthes humilis HBK cationic peroxidases. Amino acid sequence identities in the conserved domains of plant peroxidases ranged from 60 to 100%. In PpPrx75, there are five cysteine residues and one histidine residue that are found at conserved positions among other peroxidases. A potential glycosylation site (NTS) is present in the deduced sequence. Phylogenetic analysis showed that PpPrx75 is closely related to two A. thaliana peroxidases. The PpPrx75 cDNA was induced by active auxins, ethylene, abscisic acid and quercetin, a flavonoid possibly involved in plant-microorganism interactions. Transcript accumulation was detected within 3 h following root induction by auxin, and the amount of mRNA increased over the following 24 h. The protein synthesis inhibitor cycloheximide did not inhibit indole-3-acetic acid-induced transcript accumulation, suggesting that PpPrx75 induction is a primary (direct) response to auxin. This cDNA can be used to study expression of an auxin regulated peroxidase during ectomycorrhiza formation. PMID- 11874720 TI - Understanding the role of the cytoskeleton in wood formation in angiosperm trees: hybrid aspen (Populus tremula x P. tremuloides) as the model species. AB - The involvement of microfilaments and microtubules in the development of the radial and axial components of secondary xylem (wood) in hybrid aspen (Populus tremula L. x P. tremuloides Michx.) was studied by indirect immunofluorescent localization techniques. In addition to cambial cells, the differentiated cell types considered were early- and late-wood vessel elements, axial parenchyma, normal-wood fibers and gelatinous fibers, and contact and isolation ray cells. Microfilaments were rare in ray cambial cells, but were abundant and axially arranged in their derivatives once cell elongation had begun, and persisted in that orientation in mature ray cells. Microfilaments were axially arranged in fusiform cambial cells and persisted in that orientation in all xylem derivatives of those cells. Microtubules were randomly oriented in ray and fusiform cells of the cambial zone. Dense arrays of parallel-aligned microtubules were oriented near axially in the developing gelatinous fibers, but at a wide range of angles in normal-wood fibers. Ellipses of microfilaments were associated with pit development in fiber cells and isolation ray cells. Rings of co-localized microtubules and microfilaments were associated with developing inter-vessel bordered pits and vessel-contact ray cell contact pits, and, in the case of bordered pits, these rings decreased in diameter as the over-arching pit border increased in size. Although only microtubules were seen at the periphery of the perforation plate of vessel elements, a prominent meshwork of microfilaments overlaid the perforation plate itself. A consensus view of the roles of the cytoskeleton during wood formation in angiosperm trees is presented. PMID- 11874721 TI - Solute accumulation of chestnut oak and dogwood leaves in response to throughfall manipulation of an upland oak forest. AB - To determine the biochemical basis of osmotic adjustment, seasonal and treatment differences in foliar water- soluble organic solutes and inorganic ions were investigated for two hardwood species that exhibited osmotic adjustment in a Throughfall Displacement Experiment at the Walker Branch Watershed near Oak Ridge, Tennessee. Leaf samples of overstory and understory chestnut oak (Quercus prinus L.) and understory dogwood (Cornus florida L.) were collected during the 1994 (wet) and 1995 (dry) growing seasons from each of three treatments: dry ( 33% throughfall), ambient (control) and wet (+33% throughfall). Foliar soluble carbohydrates and organic acids were measured by gas chromatography-mass spectrometry. During May 1994, when the demand for sucrose was greatest, dogwood accumulated small amounts of glucose, quinic acid and Mg2+, offsetting a decline in nitrate concentration. As the mild drought continued and tree growth slowed, there was a significant accumulation of sucrose in dogwood in the dry treatment in June, and a trend toward increased K+. In overstory chestnut oak in the dry treatment over the same period, there were significant accumulations of fructose, glucose and K+, and a trend toward increased quinic acid accumulation. Sucrose did not become a key osmotically active compound in chestnut oak until August 1994. In 1995, with the exception of understory chestnut oak, there was no accumulation of K+ in either species. During the severe drought of 1995, monosaccharides, particularly glucose and fructose, accounted for most of the osmotic adjustment in both species. Among solutes, glucose constituted the largest accumulation in dogwood in the dry treatment in August 1995, followed by fructose and sucrose. There was only a moderate increase in solutes in chestnut oak trees in 1995, with fructose and glucose constituting over 50% of the predicted solute accumulation in July. Both species accumulated a wider array of solutes during the dry year than during the wet year, but treatment differences in solute accumulation in chestnut oak were partially limited by drought. The greater dehydration tolerance of chestnut oak than dogwood was evident in the higher baseline concentrations of organic solutes and the greater array of solutes accumulated in response to drought. PMID- 11874722 TI - Growth strain in coconut palm trees. AB - Until recently, growth stress studies have been made only on coniferous and dicotyledonous trees. Growth stress of trees is thought to be initiated in newly formed secondary xylem cells. This stress can accumulate for years and is distributed inside the trunk. Major characteristics of the trunk of monocotyledonous trees include numerous vascular bundles scattered inside the ground tissue and the lack of secondary growth for enlarging the diameter of the trunk. We used the strain gauge method to measure the released growth strain of the monocotyledonous woody palm, coconut (Cocos nucifera L.), and to investigate the surface growth strain of the trunk and central cylinder at different trunk heights. The internal strains of both vertical and leaning trunks were measured and compared with those of coniferous and dicotyledonous trees. We found that tensile stress existed longitudinally on the surface of vertically growing trunks, whereas compression stress was found at the bending position of leaning trunks. Compression stress was found in the outer part of the central cylinder, whereas tensile stress is generally found in the outer part of the trunk in coniferous and dicotyledonous trees. The distribution of strain in the palm trunk is similar to that of compression wood of the leaning trunk of a conifer. Specific gravity was greater in the outer part of the trunk than in the inner part of the trunk. This difference may be related to the distribution of growth stress. PMID- 11874723 TI - Rate of stomatal opening, shoot hydraulic conductance and photosynthetic characteristics in relation to leaf abscisic acid concentration in six temperate deciduous trees. AB - Correlations between leaf abscisic acid concentration ([ABA]), stomatal conductance (gs), rate of stomatal opening in response to an increase in leaf water potential (si), shoot hydraulic conductance (L) and photosynthetic characteristics were examined in saplings of six temperate deciduous tree species: Acer platanoides L., Padus avium Mill., Populus tremula L., Quercus robur L., Salix caprea L. and Tilia cordata Mill. Species-specific values of foliar [ABA] were negatively related to the mean values of gs, si, L and light- and CO2- saturated net photosynthesis (P(max)), thus providing strong correlative evidence of a scaling of foliar gas exchange and hydraulic characteristics with leaf endogenous [ABA]. In addition, we suggest that mean gs, si, L and Pmax for mature leaves may partly be determined by the species-specific [ABA] during leaf growth. The most drought-intolerant species had the lowest [ABA] and the highest gs, suggesting that interspecific differences in [ABA] may be linked to differences in species-specific water-use efficiency. Application of high concentrations of exogenous ABA led to large decreases in gs, si and P(max), further underscoring the direct role of ABA in regulating stomatal opening and photosynthetic rate. Exogenous ABA also decreased L, but the decreases were considerably smaller than the decreases in gs, si and Pmax. Thus, exogenous ABA predominantly affected the stomata directly, but modification of L by ABA may also be an important mechanism of ABA action. We conclude that interspecific variability in endogenous [ABA] during foliage growth and in mature leaves provides an important factor explaining observed differences in L, gs, si and Pmax among temperate deciduous tree species. PMID- 11874724 TI - Heat dissipation sensors of variable length for the measurement of sap flow in trees with deep sapwood. AB - Robust thermal dissipation sensors of variable length (3 to 30 cm) were developed to overcome limitations to the measurement of radial profiles of sap flow in large-diameter tropical trees with deep sapwood. The effective measuring length of the custom-made sensors was reduced to 1 cm at the tip of a thermally nonconducting shaft, thereby minimizing the influence of nonuniform sap flux density profiles across the sapwood. Sap flow was measured at different depths and circumferential positions in the trunks of four trees at the Parque Natural Metropolitano canopy crane site, Panama City, Republic of Panama. Sap flow was detected to a depth of 24 cm in the trunks of a 1-m-diameter Anacardium excelsum (Bertero & Balb. ex Kunth) Skeels tree and a 0.65-m-diameter Ficus insipida Willd. tree, and to depths of 7 cm in a 0.34-m-diameter Cordia alliodora (Ruiz & Pav.) Cham. trunk, and 17 cm in a 0.47-m-diameter Schefflera morototoni (Aubl.) Maguire, Steyerm. & Frodin trunk. Sap flux density was maximal in the outermost 4 cm of sapwood and declined with increasing sapwood depth. Considerable variation in sap flux density profiles was observed both within and among the trees. In S. morototoni, radial variation in sap flux density was associated with radial variation in wood properties, particularly vessel lumen area and distribution. High variability in radial and circumferential sap flux density resulted in large errors when measurements of sap flow at a single depth, or a single radial profile, were used to estimate whole-plant water use. Diurnal water use ranged from 750 kg H2O day-1 for A. excelsum to 37 kg H2O day-1 for C. alliodora. PMID- 11874725 TI - Pigment composition and location in honey locust (Gleditsia triacanthos) seeds before and after desiccation. AB - Gleditsia triacanthos L. pods were harvested at San Lorenzo del Escorial, Madrid, Spain, before and after seed desiccation. Maturing green pods, harvested from trees, contained soft light-green seeds, whereas fully mature brown pods, collected on the ground, contained hard dark-green seeds. Based on visual inspection of seeds, it was determined that the green color is located in the cotyledons and embryo of soft seeds but mainly in the seed coat of hard seeds. High performance liquid chromatographic analysis indicated that both hard and soft seeds contained the same set of photosynthetic pigments as fully developed leaves, but in different proportions. The hard and soft seeds mainly differed in their chlorophyll a to chlorophyll b ratio and in the composition of the xanthophyll cycle pool of pigments. Fluorescence at -196 degrees C revealed that the molecular organization of the pigment molecules in the seed coat of hard seeds differed from that in intact cotyledons of soft seeds and intact green leaves. The -196 degrees C fluorescence spectra also revealed the presence of a small heterogenous pool of non-photoactive protochlorophyll(ide)s, similar to those found in dark-grown tissues of gymnosperms and angiosperms. PMID- 11874726 TI - "Retinal imaging" aberrometry. PMID- 11874727 TI - "Retinal imaging" aberrometry. PMID- 11874729 TI - Factors in a hyperopic shift after LASIK. PMID- 11874732 TI - Posner Schlossman syndrome. PMID- 11874734 TI - Viscocanalostomy vs. trabeculectomy. PMID- 11874735 TI - Viscocanalostomy vs trabeculetomy. PMID- 11874737 TI - How good are measures of physician quality of care? PMID- 11874738 TI - Interventions to retard myopia progression in children: an evidence-based update. AB - TOPIC: To evaluate the efficacy of interventions such as eyedrops, bifocal lenses, or contact lenses in retarding the progression of myopia in myopic children. CLINICAL RELEVANCE: Myopia is a common ocular disorder, and high myopia (myopia at least -6.0 diopters) is associated with potentially blinding conditions. At present, there are no general guidelines on interventions that may decrease myopia progression in children, but some interventions such as contact lenses are offered on an ad hoc basis. METHODS OR LITERATURE REVIEWED: English and non-English language articles published from 1968 to 2000 were retrieved using a keyword search of MEDLINE, Embase, Cochrane Library, and Science Citation Index databases. Randomized controlled trials with comparisons of the effectiveness of interventions to decrease myopia progression in myopic children were reviewed. RESULTS: Ten clinical trials of different interventions to retard myopia progression were reviewed, including three trials that evaluated atropine and one trial that evaluated soft contact lenses. Atropine eye drops of 0.5% concentration were effective in clinical trials, but no significant effect was found for tropicamide or timolol eyedrops. Five of the six trials on bifocal spectacle lenses with various additions failed to show significant retardation, and results of the remaining trial were barely significant (P = 0.047). A trial of soft contact lenses failed to show significant effects. CONCLUSIONS: The latest evidence from randomized clinical trials does not provide sufficient information to support interventions to prevent the progression of myopia. Long term large-scale double-masked randomized clinical trials, including cycloplegic refraction, are needed before any recommendations about interventions in clinical practice to prevent high myopia in myopic children are considered. PMID- 11874742 TI - Human antitransforming growth factor beta(2) monoclonal antibody--a new modulator of wound healing in trabeculectomy: a randomized placebo controlled clinical study. AB - PURPOSE: The human monoclonal antibody that neutralizes the growth factor TGFbeta(2) (CAT-152) safely and effectively inhibits in vitro and in vivo models of conjunctival scarring. This phase I/IIa clinical trial was designed to assess the safety and tolerability of CAT-152 in patients undergoing trabeculectomy. DESIGN: Prospective randomized placebo-controlled clinical trial. PARTICIPANTS AND CONTROLS: Twenty-four patients who were due to undergo primary trabeculectomy at Moorfields or Western Eye Hospitals in London, England, were recruited for this study and randomly assigned to treatment with either CAT-152 (100 microg in 100 microl) (n = 16) or placebo (n = 8). METHODS: The treatment regimen was a series of four 100-microl subconjunctival injections, given immediately before and after surgery, and at 1 day and 1 week postoperatively. Assessment consisted of a full ophthalmic examination with recordings of the logarithm of the minimum angle of resolution visual acuities performed at baseline and at set intervals after surgery. Any adverse events were recorded. MAIN OUTCOME MEASURES: Logarithm of the minimum angle of resolution visual acuity, intraocular pressure, complications, and adverse events. RESULTS: The results of 12 month's follow-up on all patients are documented. There were no statistically significant differences in the incidence of complications between the two groups, and no serious adverse events related to the study drug occurred. Blebs after CAT-152 antibody treatment were diffuse, noncystic, and nonavascular, unlike blebs associated with antimetabolites. The fall in intraocular pressure was greater in the CAT-152 group at 3 and 6 months (P < 0.05) and approached statistical significance at 12 months. There was a trend toward less intervention in those patients treated with CAT-152. The small number of patients included limited the power of the study (34%) to detect a difference between groups. Sixteen patients in each arm of the study would be required to obtain a power of 90% with a 5% significance level. CONCLUSIONS: This is the first clinical study of CAT-152 in patients undergoing glaucoma filtration surgery. CAT-152 seems to be well tolerated, and based on these results further multicenter trials are underway. PMID- 11874743 TI - Baseline visual field characteristics in the ocular hypertension treatment study. AB - PURPOSE: The Ocular Hypertension Treatment Study (OHTS) seeks to evaluate the safety and efficacy of topical ocular hypotensive medication in preventing or delaying the onset of visual field loss and/or optic nerve damage in ocular hypertensive subjects at risk for developing primary open-angle glaucoma. This study evaluates the baseline visual field test characteristics (visual field status, reliability properties, etc.) of patients who underwent eligibility visual field testing for entry to the OHTS. DESIGN: Cross-sectional study of baseline data as part of a longitudinal randomized clinical trial. PARTICIPANTS: Two thousand eight hundred nineteen ocular hypertensive individuals, aged 40 to 80 (mean age, 55). METHODS: Subjects underwent at least two Humphrey Field Analyzer Program 30-2 Full Threshold visual field examinations in both eyes for study eligibility. A third examination was performed if a prior test was abnormal, questionable, or unreliable. For final eligibility, two sets of visual field examinations had to meet OHTS criteria for reliability and had to be classified as "normal." All OHTS visual field tests of potential subjects were submitted for eligibility assessment to the OHTS Visual Field Reading Center. MAIN OUTCOME MEASURES: The percentage of visual fields that were normal and reliable according to OHTS criteria. RESULTS: Of the subset of 2304 subjects who completed the eligibility assessments, 1828 (79%) were OHTS-eligible based on visual field test requirements. A third eligibility test was required for 11% of all eyes because of unreliable, questionable, or abnormal test results. With the 33% fixation loss cutoff in the OHTS, 97% of all eligibility visual field examinations were reliable and 3% were unreliable. The most frequent cause (69.5%) of unreliability was excessive fixation losses. CONCLUSIONS: Permitting one repeat test after an abnormal or unreliable test allowed an extra 560 patients to be "eligible" for the study based on visual field tests. A clinical screening review of otherwise normal and reliable tests was not restrictive. The adoption of a 33% fixation loss cutoff significantly reduced the number of required retests and prevented study rejection of 89 patients. PMID- 11874744 TI - Prevalence of the use of complementary and alternative medicine for glaucoma. AB - OBJECTIVES: To estimate the prevalence of complementary and alternative medicine (CAM) for glaucoma, explore possible demographic and disease-related associations, and inquire about the perceived benefit of these treatments. DESIGN: Cross-sectional study. SETTING AND PARTICIPANTS: One thousand twenty seven consecutive patients from two urban, referral glaucoma practices. MAIN OUTCOME MEASURE: Use of CAM specifically for glaucoma. RESULTS: The response rate was 97.4%. The percentage of people reporting use of CAM for glaucoma was 5.4% (54 of 1000 subjects) with 32 of these 54 (59%) having used more than one type. The percentages of those using the various types of nontraditional medicine were: megavitamin therapy (62.9%), herbal therapy (57.4%), exercise (24.0%), diet modification (22.2%), meditation, (1.8%), acupuncture (1.8%), faith healing (1.8%), and homeopathic remedies (1.8%). Patients who used CAM were more likely to be educated beyond high school (P = 0.0014) and less likely to be retired (P = 0.0053). Use of nontraditional therapy was not strongly associated with race (P = 0.044), age (P = 0.062), gender (P = 0.24), length of diagnosis (P = 0.91), or number of glaucoma medications (P = 0.58). Of those using nontraditional therapy, 52% believed that it was helpful, 39% were unsure, and 9% considered it not helpful; 72% discussed their use with an ophthalmologist. Seventy percent discovered it from sources other than providers of either traditional or nontraditional care. CONCLUSIONS: The prevalence of CAM use for glaucoma was 5.4% (95% confidence interval, 4.0%--6.4%). Most glaucoma patients currently cared for by ophthalmologists do not use nontraditional medicine. PMID- 11874745 TI - Standard achromatic perimetry, short wavelength automated perimetry, and frequency doubling technology for detection of glaucoma damage. AB - OBJECTIVE: Reevaluation of the relationship between short wavelength automated perimetry (SWAP), standard achromatic perimetry (SAP), and frequency doubling technology (FDT) in glaucoma and ocular hypertensive patients and in glaucoma suspects. DESIGN: Prospective comparative observational study. PARTICIPANTS: Four age-matched groups were evaluated (42 patients with early to moderate glaucoma, 34 ocular hypertensives, 22 glaucoma suspects, and 25 normal controls) using SAP, SWAP, and FDT. INTERVENTION: All participants underwent full clinical ophthalmologic evaluation followed by SWAP, SAP, and FDT perimetry within a period of not more than 3 months. Mean defect (MD), pattern standard deviation (PSD), visual field (VF) indices, and the percentage of depressed visual field points with P value <5% and <1% in the pattern deviation plot were evaluated. MAIN OUTCOME MEASURES: For each of the four study groups, the MD, PSD, and the percentage of abnormal points significantly depressed <5% and <1% were compared for the three VF testing modalities. RESULTS: In the glaucoma group, the mean percentage of the total number of abnormal points in SWAP was significantly less than abnormal points in SAP and FDT, both for 5% (P values were 0.0001 and 0.0001, respectively) and 1% (P values were 0.0001 and 0.0001, respectively). The same applied to the ocular hypertensives group. However, in the suspects group, no significant difference was detected. In normal controls, the abnormal points in SWAP were significantly lower than those in SAP for 5% (P value was 0.01) and 1% (P value was 0.05). FDT detected significantly larger defects (percentage of points <5%) than SAP in ocular hypertensives and suspects (P values were 0.01 and 0.004, respectively). CONCLUSIONS: SWAP in its existing condition is markedly less efficient than either SAP or FDT in detecting VF defects, especially in glaucoma patients and ocular hypertensives (defects detected with SWAP are less than both SAP and FDT). Defects detected with FDT are equivalent to SAP and sometimes larger, especially in ocular hypertensives and glaucoma suspects; this makes it a useful tool for picking up early glaucomatous defects in populations at risk. PMID- 11874746 TI - Change detection in regional and volumetric disc parameters using longitudinal confocal scanning laser tomography. AB - PURPOSE: To develop a clinically specific strategy for optic nerve head (ONH) surface change detection within longitudinally acquired confocal scanning laser tomographic (CSLT) images. DESIGN: Experimental study. PARTICIPANTS AND/OR CONTROLS: Twelve monkeys, each with one glaucomatous and one contralateral normal eye. MAIN OUTCOME MEASURES: Detection of ONH surface change within the CSLT images of each monkey's glaucomatous eye. METHODS: Six 10 and six 15 CSLT images and four stereo optic disc photographs were obtained from both eyes of 12 monkeys on 3 separate days (imaging sessions) and then every 2 weeks after laser to one eye (study eye) to elevate intraocular pressure. For each set of 10 and 15 images per eye (and for each of the 7-25 imaging sessions per monkey), 37 CSLT parameters were calculated. Univariate change required an individual parameter to change in excess of its analysis of variance-determined minimum detectable change (MDC) value. Multivariate change required groups of three parameters, considered together, to demonstrate significant change as determined by a multivariate analysis of variance. The rate of false-positive change detection for each individual parameter and for a group of three-parameter combinations was determined using both a one-in-a-row strategy (change at a single session) and a two-in-a-row strategy (change at two successive sessions) within the postlaser images of the 12 unchanging contralateral normal eyes. Change detection within the study eye images was then assessed for only those individual parameters and three-parameter combinations that were clinically specific (i.e., showed less than 10% false-positive change detection in the normal eyes). RESULTS: A total of 36 prelaser (three per monkey) imaging sessions and 158 postlaser (4-22/monkey) imaging sessions was performed. Clinically specific change detection (low rate of false positives) was achieved only with the two-in-a-row strategy. Overall, multivariate ONH surface change detection performed best; the best-performing three-parameter combination detected only 8 change events (4 onset and 4 progression) in 139 imaging sessions within the postlaser imaging sessions of the contralateral normal eyes and a total of 47 change events (11 onset and 36 progression) within the postlaser imaging sessions of the 12 study eyes. Counterintuitive (anterior) change occurred in most parameters within the late postlaser imaging sessions of the study eyes followed to end-stage damage. CONCLUSIONS: Clinically specific detection of the onset and progression of glaucomatous ONH surface change is possible within longitudinally acquired CSLT images. PMID- 11874747 TI - Clinician change detection viewing longitudinal stereophotographs compared to confocal scanning laser tomography in the LSU Experimental Glaucoma (LEG) Study. AB - PURPOSE: To compare optic nerve head (ONH) surface change detection by confocal scanning laser tomography (CSLT) within the LSU Experimental Glaucoma (LEG) study to expert clinicians viewing the LEG stereophotographs. DESIGN: Experimental study. PARTICIPANTS: Four fellowship-trained glaucoma specialists. METHODS: In the LEG study, six 15 CSLT images (TopSS, Laser Diagnostics Technologies, San Diego, CA) and four 2x optic disc stereophotographs were obtained from both eyes of 12 monkeys on 3 separate days and then every 2 weeks after laser to one eye (study eye) to elevate intraocular pressure. ONH surface change detection within the CSLT images is described in our companion report. In this report, the preliminary study compared change detection by the CSLT multivariate strategy with that of a single clinician viewing stereophotograph pairs on three separate occasions as the "gold standard." The main study compared change detection by three additional clinicians viewing a subset of LEG stereophotograph pairs on three separate occasions with that of the CSLT multivariate strategy as the "gold standard." Clinician change detection was assessed for partial (two of three occasions) or complete (three of three occasions) agreement. Three comparison groups within the main study are emphasized: 44 group A comparisons assessed false-positive change detection (specificity); 38 group B comparisons assessed change detection within 38 instances (11 onset and 27 progression events) of CSLT detected study eye change; and 30 group C comparisons assessed change detection within 30 instances in which the CSLT failed to detect change in study eyes. MAIN OUTCOME MEASURES: Clinician change detection within each comparison with either partial or complete agreement. RESULTS: Within the main study, intrareading (-.29 to 0.67) and interreading (0.24--0.56) session agreement for each clinician was slight to moderate by kappa test. Good specificity (less than 10% false-positive change detection) was achieved within the 44 group A comparisons by two of the three clinicians, but only when the more stringent criterion (change detection on three of three occasions) was applied. Of the 38 group B comparisons (in which the CSLT detected change), the two clinicians who achieved good specificity in group A failed to detect change in 25 and 16 instances, respectively, using the more stringent (three of three) criterion. Similarly, of the 30 group C comparisons (in which the CSLT failed to detect change), these two clinicians detected change in three and seven instances, respectively, but in only one comparison did they both detect change. CONCLUSIONS: This study provides the first direct evidence that an existing CSLT may reasonably meet or exceed the ONH surface change detection performance of fellowship-trained glaucoma specialists in at least those eyes with good CSLT reproducibility. PMID- 11874748 TI - Sleep apnea and intracranial hypertension in men. AB - PURPOSE: To investigate sleep apnea as an associated finding in idiopathic intracranial hypertension (IIH) in men. DESIGN: Multicenter, retrospective, noncomparative interventional case series. METHODS: Retrospective review of all men with the diagnosis of IIH seen within the last 5 years at three tertiary care academic ophthalmologic institutions. Cases with sleep apnea (SA) and IIH were identified and reviewed. RESULTS: Thirty-two cases of IIH in men were reviewed. Six cases with SA met the modified Dandy criteria for the diagnosis of IIH. Of these six patients, one received acetazolamide alone, four received acetazolamide and continuous positive airway pressure (CPAP), and one was treated with CPAP alone. All patients had preserved central acuity (20/20 or better in both eyes), enlarged blind spots, and optic disc edema in both eyes. Five patients had normal visual fields after treatment, and one patient had residual visual field loss. Three patients had normal optic nerve examinations, with resolution of the optic disc edema at last follow-up. After resolution of the optic disc edema, these three patients were maintained on CPAP but discontinued acetazolamide. Two patients had persistent but improved papilledema and are under continued treatment with acetazolamide and CPAP. One patient had optic disc pallor in both eyes and is stable. CONCLUSIONS: SA was a common finding in men meeting the modified Dandy criteria for IIH in adults. Treatment of sleep apnea with nocturnal oxygenation may improve the signs and symptoms of IIH in affected men. PMID- 11874749 TI - Comparing ophthalmoscopy, slide viewing, and semiautomated systems in optic disc morphometry. AB - PURPOSE: To compare disc measurements obtained by indirect ophthalmoscopy, the Heidelberg Retina Tomograph (HRT), stereoscopic slide viewing (SSV) of color transparencies, and the Topcon ImageNet System (ImageNet). DESIGN: Population based cross-sectional study. METHODS: From the Rotterdam Study, 324 subjects (567 eyes) were nonselectively included. All underwent a full ophthalmologic examination in mydriasis. Vertical cup/disc ratios (VCDRs) were compared between all four methods and disc area (mm(2)), neural rim area (mm(2)), cup area (mm(2)), and cup volume (mm(3)) between HRT and ImageNet. RESULTS: Mean VCDR for ophthalmoscopy was 0.25 (standard error [SE], 0.007), for HRT 0.42 (SE, 0.008), for SSV 0.39 (SE, 0.010), and for ImageNet 0.50 (SE, 0.006). The correlation for VCDR between ophthalmoscopy, the two devices, and SSV was 0.42, respectively 0.57; between ImageNet and HRT 0.75. The 97.5th percentiles of the VCDR for ophthalmoscopy, HRT, SSV, and ImageNet were 0.80, 0.73, 0.80, and 0.73, respectively; the 99.5th percentiles thus were 0.90, 0.79, 0.86, and 0.79. The mean disc area, rim area, cup area, and cup volume were 2.08, 1.63, 0.45 mm(2) and 0.09 mm(3) for HRT, and 2.39, 1.77, 0.61 mm(2) and 0.16 mm(3) for ImageNet, respectively. The corresponding correlations for these four parameters were 0.67, 0.42, 0.81, and 0.82. CONCLUSIONS: Different techniques lead to considerable differences in disc morphometric values. ImageNet produced higher mean values compared with HRT and ophthalmoscopy. Ophthalmoscopy showed the lowest correlations and SSV the highest ones with the two semiautomated devices. Between ImageNet and HRT the correlation for all parameters was high except for the neural rim area. PMID- 11874750 TI - Clinical features and management of intraorbital foreign bodies. AB - PURPOSE: To review the clinical features and management of patients with intraorbital foreign bodies. DESIGN: Noncomparative interventional case series. PATIENTS AND METHOD: Forty patients seen at two regional orbital surgery departments with intraorbital foreign bodies were reviewed. MAIN OUTCOME MEASURES: Visual acuity, surgical interventions, and complications. RESULTS: Seventy-three percent of patients were younger than 30 years old. There were 22 metallic, inorganic; 5 nonmetallic, inorganic; and 13 organic intraorbital foreign bodies (IOrbFb) in this series. Thirty patients were seen at the time of injury, and 10 patients were seen in a delayed setting with orbital complications. Thirty-four patients had surgical removal of their IOrbFb either because of complications or easy surgical access. Six patients had no surgery because of posteriorly located inorganic foreign bodies. Thirteen patients had resultant blind eyes; 12 of these were blind from the initial trauma. CONCLUSIONS: Loss of vision in conjunction with IOrbFbs is usually a result of the initial trauma. All patients should have antibiotic therapy because of the high incidence of secondary orbital infections. Computed tomography is the best initial mode of imaging. Surgical removal is indicated for all organic IOrbFbs. Inorganic IOrbFbs should be removed if causing complications or if located anteriorly after discussion of potential surgical complications with the patient. Posteriorly located inorganic IOrbFbs should be left alone, unless they are causing significant orbital complications. PMID- 11874751 TI - Pathogenesis of transient high myopia after blunt eye trauma. AB - OBJECTIVE: To determine the pathogenesis of transient high myopia after blunt eye trauma. DESIGN: Two observational case reports and literature review. METHODS: Refraction was measured in two patients with an autorefractometer in the acute and convalescent stages after a blunt eye injury. The anterior chamber angle, the ciliary body, and the choroid were examined by ultrasound biomicroscopy (UBM) in the acute and convalescent stages. In one patient, the anterior chamber depth, lens thickness, and axial length were measured by A-scan ultrasonography in the acute and convalescent stages. MAIN OUTCOME MEASURES: Comparison of the refraction, anterior chamber depth, lens thickness, axial length, and the UBM determined appearance of the choroid and ciliary body during the acute stage with the values during the convalescent stages. RESULTS: The first patient showed a myopic shift of -9.75 diopters (D) and an anterior chamber shallowing of 0.94 mm measured 3 days after trauma by an air bag inflation compared with the measurements at the convalescent phase. UBM showed an annular ciliochoroidal effusion with ciliary body edema, anterior rotation of the ciliary processes, and disappearance of the ciliary sulcus. Eleven days after the injury, these UBM findings normalized, and the myopia decreased to -0.75 D, 27 days after trauma. The second patient had a myopic shift of -8.9 D compared with the convalescent phase, immediately after blunt trauma by a firework. Seven days after the injury, UBM revealed a partial cyclodialysis in addition to findings similar to those in the first patient. Ten days after injury, a myopic shift (-4.75 D), anterior chamber shallowing (by 1.1 mm), and thickening of the crystalline lens (by 0.27 mm) were observed compared with the convalescent phase. Associated UBM findings confirmed the anterior shift of the lens-iris diaphragm. Seventeen days after trauma, the UBM findings, including the cyclodialysis, were normalized, and the myopia had decreased to -1.0 D. CONCLUSIONS: Transient high myopia after blunt trauma is caused by anatomic changes in the ciliary body and crystalline lens. The anterior shift of the lens-iris diaphragm caused by ciliochoroidal effusion with ciliary body edema and thickening of the crystalline lens from blunt eye trauma are involved in traumatic high myopia. PMID- 11874752 TI - Histologic findings after amniotic membrane graft in the human cornea. AB - OBJECTIVE: To describe the histopathologic findings in the human cornea several months after a stromal amniotic membrane graft. To show the clinicopathologic correlation after the graft in two cases with different follow-up times. DESIGN: Two interventional case reports with clinicopathologic correlation. PARTICIPANTS: Two patients with neurotrophic corneal ulcer unresponsive to medical treatment (one with stromal vascularization and the other without stromal vascularization). INTERVENTION: Amniotic membrane graft was performed in both patients to treat the neurotrophic ulcer. Three and 7 months after amniotic membrane grafting, a penetrating keratoplasty was needed, and the removed corneas were analyzed. MAIN OUTCOME MEASURES: Clinical and histopathologic examinations, including routine histopathologic and immunohistochemical studies. RESULTS: Complete epithelialization was observed on histologic examination over the basement membrane of the amniotic membrane graft. The amniotic membrane was slowly reabsorbed in the cornea without stromal vascularization with no inflammatory reaction produced. In the cornea that had stromal vascularization the amniotic membrane was rapidly reabsorbed because of the presence of abundant inflammatory cells. Once reabsorbed, the amniotic membrane was replaced by new fibrotic stroma, that was different from that found in the rest of the cornea but that helped to maintain corneal thickness. CONCLUSIONS: The amniotic membrane graft allows for correct epithelialization in cases of neurotrophic corneal ulcer. Once the amniotic membrane is reabsorbed, it is replaced by a new fibrotic stroma, which can reduce corneal transparency. In corneas that have no stromal vascularization, the graft may remain in the stroma for many months, compromising corneal transparency during this period. PMID- 11874753 TI - Clinical and ultrastructural features of a novel hereditary anterior segment dysgenesis. AB - OBJECTIVE: To describe the clinical, histopathologic, and hereditary features of a novel familial anterior segment dysgenesis. DESIGN: Prospective, observational case series and interventional case report. PARTICIPANTS: Ten individuals from three generations of a single family with iris and corneal abnormalities associated with congenital cataracts. MAIN OUTCOME MEASURES: An ophthalmic evaluation including slit-lamp examination, corneal topography, pachymetry, and specular biomicroscopy of all family members, and histopathologic and ultrastructural evaluation of one excised corneal button. RESULTS: The proband was an 81-year-old man with bilateral aphakia and diffuse corneal haze, and thinning associated with corneal guttae. His pupils were small, mildly eccentric, and difficult to dilate. Pachymeter readings were 335 microm (right eye) and 330 microm (left eye). Topography confirmed advanced steepening of both corneas. Light microscopic and transmission electron microscopic examinations of the corneal button revealed an attenuated endothelium with prominent intracellular random aggregates of small-diameter filaments staining positively for cytokeratin. Descemet's membrane was thickened and had marked posterior nodularity. Various-sized polymorphic vacuoles containing layered electron-dense material were present within and between collagen lamellae and within keratocytes throughout the stroma and Bowman's membrane. Secondary bullous changes of the epithelium with thickening of the basement membrane were also observed. The family pedigree demonstrated an autosomal dominant inheritance pattern. CONCLUSIONS: This constellation of autosomal dominantly inherited corneal endothelial and stromal disorder, with congenital cataracts and iris abnormalities, represents a novel anterior segment disorder. Its etiology may involve an abnormal migration of the secondary mesenchyme. PMID- 11874754 TI - Ophthalmic complications of slit-ventricle syndrome in children. AB - OBJECTIVE: To present the ophthalmic features and visual prognosis of patients with slit-ventricle syndrome (SVS). DESIGN: Observational case series. PARTICIPANTS: Six patients diagnosed and treated with SVS at the Montreal Children's Hospital between 1985 and 1999. METHODS: Patients were included in this study if they had an appropriate ophthalmologic follow-up and if they fulfilled the criteria for the diagnosis of SVS based on intracranial pressure monitoring and neuroimaging studies. MAIN OUTCOME MEASURES: Features studied included patients' baseline ophthalmologic evaluation, visual outcome, and neurosurgical characteristics. RESULTS: The follow-up ranged from 1 to 14 years. The average number of shunting procedures was 3.7. Ocular examination performed on the initial visit revealed a normal visual acuity in four patients, esotropia in three of six patients, and nystagmus in two of six patients. Cycloplegic refraction was normal in all the subjects. Initial funduscopic evaluation revealed optic atrophy in a 4-month-old infant, whereas two children developed optic atrophy later in the course of the disease. One child developed severe visual field defects. The two children with optic atrophy had moderate to severe loss of visual acuity associated with SVS. CONCLUSIONS: A prompt recognition of patients with SVS is crucial, because these individuals are at an increased risk for significant visual loss. PMID- 11874756 TI - Tactile corneal reflex development in full-term babies. AB - OBJECTIVE: To evaluate the development of the tactile corneal reflex in healthy full-term babies. DESIGN: Prospective longitudinal comparative case series. PARTICIPANTS: One hundred and ninety-one babies aged 1 to 3 days examined in the nursery, and 200 babies aged 1 to 12 weeks examined in an orthopedic outpatient clinic. METHODS: The tactile corneal reflex was evaluated by three ophthalmologists using the Cochet-Bonnet esthesiometer. MAIN OUTCOME MEASURES: The probability of the presence of the tactile corneal reflex was calculated in relation to gestational age, postpartum age, postconceptional age, and birth weight. RESULTS: The proportion of children with tactile corneal reflex was expressed in a linear logistic model. We found that 10% of the babies had tactile corneal reflex in at least one eye at 2 days of age, 25% at 1 week, 50% at 3.5 weeks, 75% at 6 weeks, and 100% at 12 weeks. The postpartum age had a greater impact on the development of the tactile corneal reflex than the gestational age. The birth weight also had a statistically significant influence (P = 0.005, Wald's test). CONCLUSIONS: The tactile corneal reflex is present in only a minority of newborns and develops during the first 3 months of life. These results demonstrate that the tactile corneal reflex has a longitudinal neurologic development and is part of the normal neurologic maturation process. PMID- 11874757 TI - Systematic unciformectomy for a standardized endonasal dacryocystorhinostomy. AB - PURPOSE: To describe a standardized osteotomy method for endonasal dacryocystorhinostomy (en-DCR) and the results obtained. DESIGN: Prospective nonrandomized, noncomparative, interventional case series study. PARTICIPANTS: One hundred consecutive adult patients (81 females, 19 males) underwent en-DCR performed by the same team of oculoplastic and rhinologic surgeons. METHODS: A standardized procedure was used in all cases. This included endoscopic assessment of the surgical site from the nasal aspect, diaphanoscopy of the common canaliculus, and endonasal osteotomy. Anterior unciformectomy was followed by resection of the lacrimal bone and protected drilling of the maxillary bone of the lacrimal fossa to expose the entire medial aspect of the lacrimal sac. The middle turbinate was preserved whenever atraumatic dissection of the nasal mucosa was possible or when it was resected partially before osteotomy. MAIN OUTCOME MEASURES: Rate of direct localization of the lacrimal fossa, rate of turbinectomy/septoplasty, rate of residual postoperative tearing, rate and nature of complications RESULTS: Transillumination of the common canaliculus was obtained directly in 94% of cases; transillumination, required the opening of an ethmoidal cell in 2%, partial resection of the middle turbinate in 3%, and unciformectomy in 1%. Access to the surgical site required partial resection of the middle turbinate in 21% of cases, but no septoplasty. Osteotomy was initiated in 90% of cases by cleavage of the anterior insertion of the uncinate process, and in 5% by cleavage of the first, overdeveloped ethmoidal cell. The cleavage opened directly to the medial aspect of the lacrimal fossa in these 95 cases. Osteotomy was achieved by drilling alone in only five cases. Perioperative complications were limited to significant bleeding in six cases. Postoperative complications included one case of resolved frontal sinusitis on day 3, and one case of inferior lid hematoma with emphysema. CONCLUSIONS: The authors suggest that anterior resection of the uncinate process is the most important surgical step to expose the medial aspect of the lacrimal fossa during endonasal DCR, whereas partial resection of the middle turbinate can be considered optional. PMID- 11874758 TI - Bilateral multifocal hemangiomas of the orbit in the blue rubber bleb nevus syndrome. AB - PURPOSE: To report the clinical findings and surgical treatment of multifocal, bilateral orbital hemangiomas in a patient with the blue rubber bleb nevus syndrome, a rare vascular disorder characterized by distinctive cutaneous and visceral hemangiomas. DESIGN: Interventional case report. METHODS: Review of clinical findings, radiologic studies, and treatment of the patient. RESULTS: A 36-year-old female with multifocal, bilateral orbital lesions was seen with severe proptosis and decreased visual acuity of the left eye. Surgical debulking of multiple cavernous hemangiomas of the orbit resulted in decreased proptosis and an improvement in visual acuity. Subsequent histopathologic analysis and the results of a systemic workup revealing multiple, distinctive, cutaneous and visceral lesions were found to be consistent with the blue rubber bleb nevus syndrome. CONCLUSIONS: Although hemangiomas of the orbit are relatively common occurrences, multifocal, bilateral lesions may exist in the setting of underlying systemic hemangiomatosis. Bilateral or multifocal hemangiomas associated with cutaneous or visceral lesions should alert the clinician to the possibility of a number of syndromes that may have life-threatening consequences. PMID- 11874759 TI - Human papillomavirus 16 and 18 expression in conjunctival intraepithelial neoplasia. AB - OBJECTIVE: To evaluate conjunctival intraepithelial neoplasia (CIN) and normal conjunctiva for the presence of human papillomavirus (HPV) DNA and for expression (as detected by the presence of mRNA) of the HPV E6 region. DESIGN: Prospective, case-controlled study. PARTICIPANTS: Ten consecutive patients who underwent CIN excision by one surgeon (CLK) and five age-matched control subjects who underwent retinal detachment repair at the Bascom Palmer Eye Institute. METHODS: A reverse transcriptase in situ polymerase chain reaction (PCR) technique was used to search for the presence of HPV mRNA in CIN specimens from 10 consecutive patients who underwent CIN excision by one surgeon (CLK) at Bascom Palmer Eye Institute, as well as in clinically uninvolved conjunctival specimens from the same eyes of these patients. In addition, conjunctival specimens from five control subjects (age-matched to five of the cases), who had no clinically identifiable conjunctival disease and who underwent retinal detachment repair at the Bascom Palmer Eye Institute, were analyzed in a similar manner. The clinical diagnoses of CIN and normal conjunctiva were confirmed histopathologically in all cases by an ocular pathologist, who was masked as to the patients' clinical diagnoses, and the PCR testing was performed by an investigator (GJN) who was masked as to the clinical diagnoses. RESULTS: HPV 16 DNA and mRNA were present in five CIN specimens, and HPV 18 DNA and mRNA were present in the other five CIN specimens; neither HPV 16 or 18 DNA nor mRNA were detected in any of the control specimens or in any of the clinically uninvolved conjunctival specimens (P < 0.001). In each of the CIN specimens, 20% to 40% of the dysplastic cells expressed the HPV E6 region. CONCLUSIONS: HPV 16 or 18 DNA and mRNA corresponding to the E6 region were detected in all CIN specimens examined. HPV 16 or 18 DNA or mRNA was not present in any of the control or uninvolved conjunctival specimens. The consistency of the current findings with those reported for human cervical malignant lesions, and the fact that the protein encoded by the E6 region of HPV 16 and 18 has been shown to form a complex with the protein encoded by the host tumor suppressor gene p53, provide strong evidence for an etiologic role of HPV in the development of CIN. PMID- 11874760 TI - Topical mitomycin-C for partially excised conjunctival squamous cell carcinoma. AB - PURPOSE: To evaluate the efficacy of topical mitomycin-C (MMC) for treatment of postoperative residual conjunctival squamous cell carcinoma (SCC). DESIGN: Retrospective noncomparative case series. PARTICIPANTS: Five patients, two males and three females, with conjunctival and histologically proven incompletely excised conjunctival SCC. METHODS: Patients were treated with topical MMC. Two to three courses of topical MMC, 0.02% or 0.04%, were applied four times daily for 14 days per course. One month after the final treatment, the scar area with surrounding normal conjunctiva was excised, and histologic evaluation was done. MAIN OUTCOME MEASURES: No evidence of malignant cells in excised tissues. RESULTS: Histologic evaluation of the five specimens showed no malignant cells. Conjunctival scarring with inflammatory response was observed. No regrowth was reported during the follow-up period of 18 to 37 months. The complications of MMC use included mild to moderate conjunctival hyperemia in three patients. All signs and symptoms were resolved after discontinuation of the treatment. CONCLUSIONS: Application of topical MMC is an efficient treatment for residual conjunctival SCC. Longer follow-up is required to confirm these findings. PMID- 11874761 TI - Histiocytoid variant of eccrine sweat gland carcinoma of the eyelid and orbit: report of five cases. AB - OBJECTIVE: To study the clinicopathologic features of the histiocytoid variant of adenocarcinoma of the eccrine sweat gland of the eye and orbit. DESIGN: Retrospective case series. PARTICIPANTS: Five patients undergoing orbital and eyelid biopsy as a diagnostic procedure. METHODS: The authors examined the clinical histories and pathologic findings of five patients with eccrine adenocarcinoma of the eyelid with orbital invasion. MAIN OUTCOME MEASURES: Clinical and histopathologic examinations, including routine histopathology, immunohistochemistry, and electron microscopy studies. RESULTS: The tumors presented as insidious, diffusely infiltrative, firm cutaneous masses in the periocular area that later infiltrated the orbit. Histopathologic examination revealed that the tumors infiltrated the dermis and were composed of cells with a histiocytic to signet ring appearance. Tumor cells exhibited intracellular mucin production. Immunohistochemical stains were positive in tumor cells for low and high molecular weight cytokeratins, carcinoembryonic antigen, and epithelial membrane antigen. Electron microscopic examination showed lumen formation and intracytoplasmic mucin in tumor cells. CONCLUSIONS: The histiocytoid variant of adenocarcinoma of the eccrine sweat gland of the eyelid may present as an insidious tumor and diffusely invade the orbit. These cases may be confused with metastatic adenocarcinoma. PMID- 11874762 TI - Detection using the multifocal electroretinogram of mosaic retinal dysfunction in carriers of X-linked retinitis pigmentosa. AB - PURPOSE: To examine whether a mosaic pattern of retinal dysfunction in obligate carriers of X-linked retinitis pigmentosa (XLRP) could be observed in local electroretinographic responses obtained with the multifocal electroretinogram (mfERG). DESIGN: Prospective observational case series. PARTICIPANTS: Five obligate carriers of XLRP (mean age, 53.2 years) were recruited into the study. METHODS: Examination of each subject included a complete ocular examination, Humphrey visual field, standard full-field electroretinogram (ERG), and mfERG testing. For the mfERG, we used a 103-scaled hexagonal stimulus array that subtended a retinal area of approximately 40 in diameter. The amplitudes and implicit times in each location for the mfERG was compared with the corresponding value determined for a group of normally sighted, age-corrected control subjects. MAIN OUTCOME MEASURES: Mapping of 103 local electroretinographic response amplitudes and implicit times within the central 40 with the multifocal electroretinogram. RESULTS: Localized regions of reduced mfERG amplitudes and/or delayed implicit times were found in four of five carriers. In one of these four carriers, a mosaic pattern of mfERG dysfunction was present even in the absence of any clinically apparent retinal changes, retinal sensitivity losses on Humphrey field testing, or abnormal full-field cone ERG responses. However, one carrier with a typical tapetal-like reflex demonstrated no deficit on any functional tests. CONCLUSIONS: The mfERG demonstrated patchy areas of retinal dysfunction in some carriers of XLRP. This mosaic pattern of dysfunction may be observed in some patients with a normal-appearing fundus, normal psychophysical thresholds, and normal amplitude and implicit time full-field ERG cone responses. PMID- 11874763 TI - Posterior segment changes associated with posterior microphthalmos. AB - PURPOSE: To characterize and analyze the posterior segment ocular involvement in patients with posterior microphthalmos. DESIGN: Retrospective observational case series. PARTICIPANTS: Eighteen patients (8 sporadic cases and 10 siblings from 5 different families) between the age of 4 and 36 years with posterior microphthalmos. METHODS: Records of patients with posterior microphthalmos over a 5-year-period were reviewed, including clinical, fundus photographic, fluorescein angiographic, and ultrasonographic findings, and management. RESULTS: All patients had bilateral foreshortening of the posterior ocular segment (range, 7- 11.2 mm) with associated high hyperopia (range, +12.00--+19.00 diopters) and normal or slightly smaller than normal anterior segment dimensions. Visual acuity ranged from 20/200 to 20/40. Inheritance of this syndrome was compatible with an autosomal recessive pattern. Posterior segment changes included bilateral elevated papillomacular retinal fold (13 patients, 72.2%); fine retinal folds (6 patients, 33.3%); chorioretinal folds (11 patients, 61.1%); uveal effusion syndrome (3 patients, 16.7%); pigmentary retinopathy (4 patients, 22.2%), including retinitis punctata albescens in 1 patient; absence or marked reduction of the capillary-free zone (18 patients, 100%); crowded optic discs (18 patients, 100%); and sclerochoroidal thickening on ultrasonography (18 patients, 100%). Two patients with uveal effusion were successfully treated with scleral surgery. CONCLUSION: A wide variety of congenital or acquired posterior segment changes may be encountered in patients with posterior microphthalmos. Although high hyperopia and elevated papillomacular retinal fold are the main causes of visual impairment, other chorioretinal changes, such as pigmentary retinopathy, chorioretinal folds and uveal effusion syndrome, should be considered as causes of visual disturbance in patients with posterior microphthalmos. Early ultrasonographic diagnosis, close follow-up, and appropriate management are mandatory to improve or maintain visual function in such patients. PMID- 11874764 TI - A distinctive form of congenital stationary night blindness with cone ON-pathway dysfunction. AB - PURPOSE: To characterize a distinctive form of congenital stationary night blindness (CSNB). DESIGN: Observational case report. PARTICIPANTS: A 30-year-old male with a history of night blindness, several members of his family, a patient with "complete" congenital stationary night blindness (CSNB1), and groups of age similar control subjects. METHODS: Rod-system function was evaluated by measuring psychophysical dark-adapted thresholds, by recording dark-adapted electroretinograms (ERGs), and by fundus reflectometry. Cone-system function was evaluated by recording light-adapted ERGs, including those to sawtooth flicker, and by recording light-adapted visually evoked potentials (VEPs) to luminance increments and decrements. MAIN OUTCOME MEASURES: Dark-adapted thresholds, ERGs, rhodopsin double densities, Goldmann visual fields, and VEPs. RESULTS: The patient's visual acuity, visual fields, and color vision were normal. His peripheral dark-adapted thresholds were rod-mediated but elevated by approximately 3 log units above normal. Rhodopsin double density and bleaching recovery were normal. His dark-adapted maximal-flash ERG showed a "negative" waveform, in which the b-wave was more reduced in amplitude than the a-wave, although the a-wave amplitude was also reduced. The rod photoreceptors contributed to the patient's dark-adapted ERGs, as illustrated by the unequal responses to cone-matched stimuli. The patient's cone-mediated thresholds for long-wavelength stimuli were within the normal range. However, his light-adapted brief-flash b-wave was abnormal in amplitude and implicit time. Selective abnormalities of the ON responses of the cone system were apparent in the patient's reduced b-wave amplitude to rapid-on flicker with a normal response to rapid-off flicker, and his prolonged VEP latencies to increments but not to decrements. CONCLUSIONS: The overall pattern of findings distinguishes this patient from previously described forms of CSNB. The results suggest that two factors likely contribute to the patient's night blindness: (1) a rod phototransduction defect and (2) a postreceptoral defect. The results also indicate dysfunction within the cone ON pathway. PMID- 11874765 TI - Sildenafil-associated nonarteritic anterior ischemic optic neuropathy. AB - PURPOSE: To describe the clinical features of five patients who developed nonarteritic anterior ischemic optic neuropathy (NAION) after ingestion of sildenafil citrate (Viagra; Pfizer Pharmaceuticals, New York, NY). DESIGN: Retrospective observational case series. PARTICIPANTS: Five patients with NAION who reported the use of sildenafil citrate before the onset of ocular symptoms. MAIN OUTCOME MEASURES: The symptoms presented, history, ophthalmic examination, and visual field examination of each patient. RESULTS: Nonarteritic anterior ischemic optic neuropathy developed in one eye within minutes to hours after ingestion of sildenafil. Four of the five patients had no vascular risk factors for ischemic optic neuropathy. The patients all developed unilateral blurry vision, altitudinal visual field defects, and optic disc edema. Each of the patients was noted to have a small cup-to-disc ratio in the unaffected optic nerve. CONCLUSIONS: Sildenafil citrate may be associated with NAION. A small cup to-disc ratio may be a risk factor for development of NAION in association with the use of sildenafil. PMID- 11874766 TI - Noninvasive diagnosis and ophthalmic features of mucolipidosis type IV. AB - OBJECTIVE: To comprehensively describe the ophthalmic characteristics of patients with mucolipidosis type IV. DESIGN: Prospective natural history study. PARTICIPANTS: Twenty-two patients with confirmed mucolipidosis type IV. METHODS OR TESTING: External and slit-lamp examination with dilated funduscopy, photography of corneal and retinal lesions, and exfoliative conjunctival cytology were performed. MAIN OUTCOME MEASURES: Grading of corneal, optic nerve, retinal vasculature, and pigmentary abnormalities. RESULTS: All patients exhibited some degree of corneal epithelial haze, optic nerve pallor, retinal vascular attenuation, and retinal pigment epithelial changes. The associated ocular findings observed in decreasing order of frequency were strabismus, corneal erosion, cataract, corneal abnormalities, fundus abnormalities, and ptosis. The older patients were significantly more likely to demonstrate severe optic nerve pallor, retinal vascular attenuation, and corneal epithelial haze. Conjunctival cytologic studies showed characteristic lysosomal inclusions on light and electron microscopy. CONCLUSIONS: Patients with mucolipidosis type IV have characteristic ophthalmic features, most of which have a progressive course. Conjunctival cytologic studies help confirm the diagnosis of this disorder. PMID- 11874767 TI - Clinical evaluation of patients with diabetic retinopathy: accuracy of the Inoveon diabetic retinopathy-3DT system. AB - PURPOSE: This study analyzed the accuracy of the Inoveon Diabetic Retinopathy (DR 3DT) system (Inoveon Corp., Oklahoma City, OK), a scalable evaluation method for the management of diabetic retinopathy using high-quality digital retinal imaging. DESIGN: An independent, masked, cross-sectional, clinical validation study. PARTICIPANTS: Two hundred ninety adult patients with diabetes from the Chickasaw Nation's Carl Albert Indian Health Facility in Ada, Oklahoma. METHODS: All participants underwent DRS7 imaging using a Zeiss FF450 fundus camera with images recorded on 35-mm film and a Kodak DCS520 digital camera back. Masked double grading with independent third reader adjudication yielded an Early Treatment Diabetic Retinopathy Study (ETDRS) Final Retinopathy Severity Scale Level (ETDRS Level) and macular edema stage for each eye. The presence of greater than-or-equal ETDRS Level 53, questionable or definite clinically significant macular edema in either eye, or ungradeable images was defined as a threshold event requiring referral. MAIN OUTCOME MEASURES: Accuracy (sensitivity, specificity, predictive values) of the digital system relative to the film "gold standard" on the threshold referral criteria per patient. RESULTS: All patients with gradeable 35-mm slides from at least one eye were included in this per patient analysis (n = 290). The prevalence of threshold events was 19.3%. The sensitivity of the digital system in detecting threshold events was 98.2% (95% confidence interval [CI], 90.5%-100.0%) and specificity 89.7% (95% CI, 85.1% 93.3%). The positive predictive value was 69.5% and negative predictive value 99.5% for this sample. CONCLUSIONS: When compared with the "gold standard," Inoveon's DR-3DT system provides highly accurate diabetic retinopathy referral decisions. Given their inherent advantages, high-quality digital imaging systems could replace the film "gold standard" as the basis for scalable, accessible, diabetic retinopathy evaluation. PMID- 11874768 TI - Hyperopic laser-assisted in situ keratomileusis for radial keratotomy induced hyperopia. AB - PURPOSE: To evaluate hyperopic laser in situ keratomileusis (H-LASIK) for radial keratotomy (RK)-induced hyperopia. DESIGN: Noncomparative interventional retrospective nonconsecutive case series. PARTICIPANTS: Sixty-nine eyes of 47 patients who had undergone RK and were seen with induced hyperopia. METHODS: H LASIK was performed with an excimer laser. MAIN OUTCOME MEASURES: The mean refractive error, in spherical equivalents (SE), uncorrected visual acuity (UCVA), and best spectacle-corrected visual acuity (BSCVA) before and after H LASIK are reported. Safety was analyzed using a mean follow-up time of 6.6 plus minus 3.24 months. RESULTS: Preoperative mean SE was +3.4 plus minus 1.6 diopters (D). Postoperative mean SE was -0.32 plus minus 1.2 D. A high percentage of eyes (79.7%; n = 55) were between plus minus1.0 D of emmetropia and 88% within plus minus 2.0 D. Preoperative BSCVA was 20/20 in 53.6% of eyes (n = 37) and 20/40 or better in 100% (n = 69). Postoperative BSCVA was 20/20 in 55% of eyes (n = 38) and 20/40 or better in 95.6% (n = 66) of eyes. Preoperative UCVA was less-than-or equal20/50 in 52 cases (75.4%). Postoperative UCVA was 20/20 in 13 cases (18.8%) and greater-than-or-equal20/40 in 45 cases (65.2%). Four eyes lost 2 Snellen lines because of epithelial ingrowth in the interface (n = 3) and diffuse lamellar keratitis (Sands of the Sahara syndrome; n = 2). One of the eyes with Sahara syndrome also had epithelial ingrowth and flap necrosis. Thirteen eyes lost 1 Snellen line, and 50 eyes maintained or gained Snellen lines. The only intraoperative complication was incision opening (n = 8) while the flap was lifted; there were no further complications. These patients did not lose any Snellen lines of their BSCVA. CONCLUSIONS: H-LASIK can be used successfully to correct RK-induced hyperopia. PMID- 11874769 TI - Incremental cost-effectiveness of initial cataract surgery. AB - PURPOSE: The purpose of this study was to perform a reference case, cost-utility analysis of initial cataract surgery using the current literature on cataract outcomes and complications. DESIGN: Computer-based econometric modeling. METHODS: Visual acuity data of patients treated and observed over a 4-month postoperative period was obtained from the US National Cataract Patient Outcomes Research Team (PORT). The results from this prospective study were combined with other studies that investigated the complication rates of cataract surgery to complete the cohort of patients and outcomes. These synthesized data were incorporated with time-tradeoff utility values, decision analysis, and econometric modeling to account for the time value of money. MAIN OUTCOME MEASURES: The number of quality adjusted life-years (QALYs) gained was calculated for the study group undergoing cataract extraction in the first eye when the vision was the same in both eyes. This was divided into the cost of the procedure to find the year 2000 nominal US dollars spent per quality-adjusted life-year ($/QALY) gained. RESULTS: Initial cataract surgery, compared with observation, resulted in a mean gain of 1.776 QALYs per patient treated. A 3% annual discount rate was used to account for the benefit over time, yielding 1.25 QALYs gained. The mean cost of treatment (also discounted at a 3% annual rate) of each patient totaled 2525 US dollars. The cost divided by the discounted benefit resulted in $2020/QALY gained for this procedure. CONCLUSIONS: Initial cataract surgery seems to be highly cost effective compared with procedures across multiple medical specialties. This information, incorporating patient preferences into evidenced-based medicine, will play an increasingly important role in the evaluation of health care in the future. PMID- 11874771 TI - Peer and expert opinion and the reliability of implicit case review. AB - OBJECTIVE: To compare the interrater and intergroup agreement in judging physician maloccurrence and compliance with standards of care using the implicit case review process. DESIGN: Mail survey with questionnaire. PARTICIPANTS: Case reviews and questionnaires were mailed to 140 board-certified ophthalmologists and 140 board-certified ophthalmologists with fellowship training. MAIN OUTCOME MEASURE: Agreement judging maloccurrence and compliance with standard of care within each group and between general ophthalmologists and specialists. RESULTS: Ninety-seven (35%) questionnaires were returned. Overall, 35% of respondents believed that ophthalmologists in the case reviews committed an error of either commission or omission. Forty-five percent of reviewers believed that physicians did not meet the standard of care. There was good within-group agreement for finding clinical error in management and not meeting the standard of care for all groups (kappa coefficient range: 0.55-0.83; P = < 0.004) except retina specialists (kappa coefficient = 0.12; P = 0.2). CONCLUSIONS: Unstructured implicit case review is not a reliable method for determining physician error or for measuring compliance with standards of care. The process is susceptible to bias, and results may vary with reviewer background or training. Unstructured implicit case review needs to be regarded as a rough screening tool and used accordingly. PMID- 11874772 TI - Hypertonic sodium bicarbonate for Taxus media-induced cardiac toxicity in swine. AB - OBJECTIVE: To determine whether intravenous (IV) hypertonic sodium bicarbonate is effective in the reversal of QRS widening associated with severe Taxus intoxication. METHODS: Seventeen anesthetized and instrumented swine were poisoned with an IV extract of Taxus media until doubling of the QRS interval on electrocardiography was achieved. After poisoning (time zero), the animals received either 4 mL/kg IV 8.4% sodium bicarbonate (experimental group; 6 animals), a similar volume of 0.7% NaCl in 10% mannitol (mannitol group; 6 animals), or nothing (control group; 5 animals). The main outcome parameter was QRS duration. Secondary outcome parameters were mean arterial pressure (MAP), heart rate (HR), and cardiac index (CI = cardiac output/kg). Additionally, arterial pH, partial pressure of carbon dioxide (pCO(2)), and plasma-ionized calcium, sodium, and potassium were monitored. RESULTS: Taxus toxicity, defined as a 100% increase in QRS duration, was produced in all animals. The animals were similar in regard to baseline and time 0 physiologic parameters as well as amount of Taxus media extract administered. From times 5 through 30 minutes, following assigned treatment, significant increases in QRS duration were detected in the experimental and mannitol groups compared with the control group. A significant lowering of MAP was found in the experimental group compared with the control group. No significant difference between groups was noted in HR or CI. The swine treated with hypertonic sodium bicarbonate had a statistically significant increase in pH, plasma sodium concentration, and base excess compared with the other groups. CONCLUSIONS: Hypertonic sodium bicarbonate was ineffective in reversing the widening of QRS interval associated with Taxus poisoning in this swine model. PMID- 11874773 TI - Determination of left ventricular function by emergency physician echocardiography of hypotensive patients. AB - OBJECTIVE: To determine whether emergency physicians (EPs) with goal-directed training can use echocardiography to accurately assess left ventricular function (LVF) in hypotensive emergency department (ED) patients. METHODS: Prospective, observational study at an urban teaching ED with >100,000 visits/year. Four EP investigators with prior ultrasound experience underwent focused echocardiography training. A convenience sample of 51 adult patients with symptomatic hypotension was enrolled. Exclusion criteria were a history of trauma, chest compressions, or electrocardiogram diagnostic of acute myocardial infarction. A five-view transthoracic echocardiogram was recorded by an EP investigator who estimated ejection fraction (EF) and categorized LVF as normal, depressed, or severely depressed. A blinded cardiologist reviewed all 51 studies for EF, categorization of function, and quality of the study. Twenty randomly selected studies were reviewed by a second cardiologist to determine interobserver variability. RESULTS: Comparison of EP vs. primary cardiologist estimate of EF yielded a Pearson's correlation coefficient R = 0.86. This compared favorably with interobserver correlation between cardiologists (R = 0.84). In categorization of LVF, the weighted agreement between EPs and the primary cardiologist was 84%, with a weighted kappa of 0.61 (p < 0.001). Echocardiographic quality was rated by the primary cardiologist as good in 33%, moderate in 43%, and poor in 22%. The EF was significantly lower in patients with a cardiac cause of hypotension vs. other patients (25 +/- 10% vs. 48 +/- 17%, p < 0.001). CONCLUSIONS: Emergency physicians with focused training in echocardiography can accurately determine LVF in hypotensive patients. PMID- 11874774 TI - Emergency medical preparedness and response to a Singapore airliner crash. AB - A Singapore airline crash in 2000 was the first documented mass casualty incident (MCI) caused by an aircraft disaster in Taiwan. This report reviews the emergency medical preparedness of the airport and examines its effects on the medical response during this incident. The anticipated benefits from the new MCI plan and prior medical preparedness were not achieved during this disaster. Poor compliance with the new MCI plan by the airport authority was noted. Victims were not triaged and did not receive adequate field medical care. Structural problems with the MCI plan were also noted. Site medical teams responding from hospitals could not function as the plan had been designed. This article discusses factors causing the poor compliance, various problems, and related issues in medical response to the incident. As learned from this experience, a properly and practically designed MCI plan, good compliance of responders, and a strong support system of responding agencies are the most important factors for successful emergency response to any MCI. PMID- 11874775 TI - Assessment of clinically significant changes in acute pain in children. AB - OBJECTIVE: To quantify, using two pain assessment scales, the amount of change in pain severity required to achieve a clinically significant improvement in pain in children presenting to a pediatric emergency department (ED) with pain. METHODS: Prospective, descriptive study involving all children presenting to a pediatric ED between 5 and 16 years of age inclusive with acute pain. Children were excluded if they 1) were intoxicated or had altered sensorium, 2) were clinically unstable, 3) were non-English-speaking, or 4) were developmentally delayed. Written informed consent was obtained. Children were asked to mark their current pain severity on the standardized Color Analogue Scale (CAS) and Faces Pain Scale (FPS). After each pain control intervention the child was asked to repeat these measurements and to describe whether his or her pain was "much less," "a little less," "about the same," "a little worse," or "much worse" compared with before. This process was repeated until the child was discharged from the ED or had a score of zero. The main outcome measure was the smallest change on the CAS or FPS necessary to cause the child to describe his or her pain as a "little less." This was defined as the clinically significant change in pain. The "ideal" change in pain was defined as the amount of change necessary for the child to describe the pain as "much less" or at which point the child thought he or she no longer required any medicine to help the pain go away. RESULTS: One hundred twenty-one children were enrolled with a mean age of 9.8 years (SD +/- 3.15). Males accounted for 56%. Pain was traumatic in 65% and nontraumatic in 35%. A total of 153 pain comparisons were made using the CAS and 154 using the FPS. Only three children complained that their pain got worse (two a little worse and one much worse). Pain was described as "the same" in 20. Of the 60 pain comparisons judged to be a "little less," the CAS score changed by a median of 2.0 cm [interquartile ratio (IQR) 1-3], and the FPS by 1.0 face (IQR 1-2). In the 71 children who judged their pain to be "much less," the CAS decreased by a median of 4.0 cm (IQR 2-5) and the FPS by 2.0 faces (IQR 2-3). CONCLUSIONS: The assessment and treatment of pain in children are an important component of pediatric practice, especially in the ED. This study provides health care professionals and clinical investigators the information necessary to assess whether their method of pain control in children is clinically relevant. PMID- 11874777 TI - The epidemiology and diagnosis of penetrating eye injuries. AB - It is estimated that there are 3.1 penetrating eye injuries per 100,000 person years in the United States. OBJECTIVES: To evaluate the epidemiology of penetrating eye injuries and to identify physical examination findings that facilitate the diagnosis and ophthalmologic referral of patients with these injuries. METHODS: This was a retrospective chart review of emergency department patients with penetrating eye injuries seen for evaluation from July 1987 to January 1999. The setting was a tertiary referral, university hospital. Three hundred eighty-four patients with 390 penetrating eye injuries were enrolled; 56% were transferred from outlying hospitals. RESULTS: Penetrating eye injuries were seen almost three times per month. Eighty percent of the injuries occurred in males, and the mean age was 29 years. Twenty-five percent of the patients had used alcohol in the period immediately preceding the injury. Final visual outcome was 28% with enucleation, "no light perception" (NLP) in 10%, light perception to 20/200 in 24%, and light perception of 20/200 or better in 38%. Poor visual outcome was associated with poor initial visual acuity, alcohol use, and delayed presentation (p = 0.036, 0.025, 0.036, respectively). Gun-related injuries caused 33% and motor vehicle crashes (MVCs) caused 21% of the worst outcomes (enucleation or NLP). In MVCs where seat belt use was reported, 71% of injured patients were unrestrained. The most common initial physical findings were hyphema (76%), abnormality of the pupil or uvea (94%), and initial visual acuity worse than 20/200 (77%). All patients had at least one of these findings. Complications occurred in 25% of cases, most commonly traumatic cataract or infection. Complications occurred more commonly in those patients transferred than in those presenting directly (p = 0.002). CONCLUSIONS: Penetrating eye injuries are relatively common, occur predominantly in young males, and often result in poor visual outcome in the affected eye. Motor vehicle crashes, alcohol use, and fire-arm use are associated with more severe injuries. PMID- 11874776 TI - How useful are clinical features in the diagnosis of acute, undifferentiated chest pain? AB - OBJECTIVES: To measure the predictive value and diagnostic performance of clinical features used to diagnose coronary syndromes in patients presenting with acute, undifferentiated chest pain. METHODS: The clinical features of patients presenting to the authors' chest pain unit with acute, undifferentiated chest pain were prospectively recorded on a standard form. Admitted patients were followed up by case note review. Discharged patients were followed up as outpatients three days later. Six months after the emergency department visit, evidence of adverse events was searched for from the hospital computer database, case notes, and the patient's primary care physician. The authors tested the power of each feature to predict: 1) acute myocardial infarction (AMI) by World Health Organization criteria, and 2) any acute coronary syndrome (ACS), evidenced by cardiac testing, AMI, arrhythmia, death, or revascularization procedure within six months. RESULTS: Eight hundred ninety-three patients were assessed, 34 (3.8%) with AMI and 81 (9.1%) with ACS. Features useful in the diagnosis of AMI were exertional pain [likelihood ratio (LR) = 2.35], pain radiating to the shoulder or both arms (LR = 4.07), and chest wall tenderness (LR = 0.3). Features useful in the diagnosis of ACS were exertional pain (LR = 2.06) and pain radiating to the shoulder, the left arm, or both arms (LR = 1.62). The site or nature of pain and the presence of nausea, vomiting, or diaphoresis were not predictive of AMI or ACS. CONCLUSIONS: Important differences exist when clinical features are specifically investigated in patients with acute chest pain and a nondiagnostic electrocardiogram. Clinical features have a limited role to play in triage decision making. PMID- 11874778 TI - Reflections on resilience. PMID- 11874787 TI - Emergency preparedness: the World Trade Center and Singapore airline disasters. PMID- 11874788 TI - Ethics seminars: physician complicity in the Holocaust: historical review and reflections on emergency medicine in the 21st century, part I. AB - Individual physicians as well as the medical establishment were complicit in a wide range of activities carried out by the Nazis during the period that encompassed the Holocaust. This article examines these activities and lists eight moral failures attributable to physicians of this era. The accompanying article reviews the ethical pitfalls encountered by German physicians during the Nazi era and examines them in relationship to current issues. It also explores the role of professionalism then and now. In particular, ethical issues presently confronting emergency physicians are examined through this prism. PMID- 11874789 TI - Ethics seminars: physician complicity in the Holocaust: historical review and reflections on emergency medicine in the 21st century, part II. AB - Part I of this seminar in ethics reviewed the participation of German physicians and the German medical establishment in carrying out Nazi policies and listed eight moral failures that could be attributed to doctors during the dark period of history known as the Holocaust. The collective acts that occurred during this period have, arguably, become a benchmark for abject ethical collapse on the part of mankind. Part II contemplates a variety of contemporary issues through the prism of the Holocaust. This article reviews and categorizes ethical pitfalls encountered by physicians during the Nazi era and examines them in relationship to several current issues. It also focuses on ethical concerns and challenges that confront contemporary emergency practitioners, some of which have parallels, though certainly not direct comparators, in the Nazi era. PMID- 11874790 TI - Social bias and injustice in the current health care system. AB - This paper reflects upon historical and modern events and challenges emergency physicians to affirm a genuine commitment to social justice. Such an affirmation does not allow the physician to rest in the belief that the system is inherently just. Rather, it challenges the practitioner to recognize the widespread and inherent injustices that are present. It is probable that significant strides have been made toward protecting the rights and dignity of our patients. Even so, much remains to be done. Poor and minority patients are still less than optimally treated, and increasing marketplace competitiveness may jeopardize some of the recent gains in caring for the uninsured. Future generations may look upon some of the current discriminatory practices of our professional lifetimes with the condemnation that we hold for past abuses. PMID- 11874791 TI - Filming of patients in academic emergency departments. AB - With increasing availability and utilization of advanced technologic modalities in medicine, questions frequently arise regarding the appropriate use of recorded images of patients. While recorded images (photography, video, etc.) of patients may often be appropriate for documentation, medical record use, peer review, and teaching, the nonmedical use of recorded images for entertainment or commercial purposes is more problematic, both ethically and procedurally. Practices regarding filming of patients in academic emergency departments are reviewed, and suggested guidelines are provided regarding the appropriate and inappropriate filming of patients. PMID- 11874793 TI - Mission-based budgeting hours model: an accurate way to understand physician costs. AB - Financial resources to manage academic emergency departments are becoming more scarce. The Association of American Medical Colleges (AAMC) has embraced mission based management (MBM) as one viable option to allocate financial and personnel resources based on medical school goals. The key features of this style are integrating financial statements, measuring faculty activity, holding service line leaders accountable, and building a trust through information sharing. The authors describe how they integrated their service line cost through an hours model. From this they describe a formula for calculating the cost of protected time. The authors use mission-based budgeting to more accurately understand their physician costs and use the formula to effectively negotiate internal and external contracts. PMID- 11874794 TI - The day that the START triage system came to a STOP: observations from the World Trade Center disaster. AB - The Fire Department of the City of New York--Emergency Medical Services (EMS) Operations is one of the largest EMS systems in the country. On a daily basis, this system responds to approximately 3,000 to 3,500 calls for ambulance requests. This equates to 1.2 to 1.5 million calls annually. As part of its response, EMS deals on a daily basis with multiple casualty and disaster-type situations. The magnitude of the attacks on the World Trade Center, however, was on a scale not previously seen by any system. This article is a case report of the September 11, 2001, incident. PMID- 11874795 TI - Is potential HIV exposure considered to be a medical emergency? PMID- 11874797 TI - Filming patients without prior consent. PMID- 11874798 TI - Filming patients without prior consent. PMID- 11874799 TI - Filming patients without prior consent. PMID- 11874802 TI - "I don't know what you guys are measuring but you sure are measuring it!" A fair criticism of measurements of exhaled condensates? PMID- 11874804 TI - Pesticide exposure and asthma. PMID- 11874803 TI - Sleep-disordered breathing and the current epidemic of obesity: consequence or contributing factor? PMID- 11874805 TI - Critical care medicine in AJRCCM 2001. PMID- 11874806 TI - Sleep-disordered breathing, control of breathing, respiratory muscles, and pulmonary function testing in AJRCCM 2001. PMID- 11874807 TI - Asthma, airway biology, and nasal disorders in AJRCCM 2001. PMID- 11874808 TI - Pediatrics, surfactant, and cystic fibrosis in AJRCCM 2001. PMID- 11874809 TI - Tuberculosis, lung infections, interstitial lung disease, and socioeconomic issues in AJRCCM 2001. PMID- 11874810 TI - Chronic obstructive pulmonary disease, pollution, pulmonary vascular disease, transplantation, pleural disease, and lung cancer in AJRCCM 2001. PMID- 11874811 TI - Dilution of respiratory solutes in exhaled condensates. AB - Most exhaled water is produced as gaseous water vapor, which can be collected in cooled condensers. The presence of nonvolatile solutes in these condensates suggests that droplets of respiratory fluid (RF) have also been collected. However, calculation of RF solute concentrations from condensates requires estimation of the dilution of RF droplets by water vapor. We used condensate electrolyte concentrations to calculate the dilution of RF droplets in condensates from 20 normal subjects. The total ionic concentration (conductivity) was 497 plus minus 68 (mean plus minus SEM) muM. Of this, 229 plus minus 43 muM was NH(4)(+), but little NH(4)(+) was collected from subjects with tracheostomies, indicating oral formation. The Na+ concentration in condensate ([Na+](cond)) averaged 242 plus minus 43 muM. Large variations in [Na(+)](cond) correlated well with variations of K+ in condensate ([K+](cond)) and Cl-) in condensate ([Cl-](cond)), and were attributed to differences in respiratory droplet dilution. Dividing condensate values of ([Na+] + [K+] ) by those of plasma indicated that RF represented between 0.01% and 2.00% of condensate volumes. Calculated values for Na+, K+, Cl-, lactate, and protein in RF were [Na+](RF) = 91 +/- 8 mM, [K+](RF) = 60 +/- 11 mM, [Cl-](RF) = 102 +/- 17 mM, [lactate](RF) = 44 +/- 17 mM, and [protein](RF) = 7.63 +/- 1.82 g/dl, respectively. PMID- 11874812 TI - Obstructive sleep apnea is independently associated with insulin resistance. AB - Epidemiological studies have implicated obstructive sleep apnea (OSA) as an independent comorbid factor in cardiovascular and cerebrovascular diseases. It is postulated that recurrent episodes of occlusion of upper airways during sleep result in pathophysiological changes that may predispose to vascular diseases. Insulin resistance is a known risk factor for atherosclerosis, and we postulate that OSA represents a stress that promotes insulin resistance, hence atherogenesis. This study investigated the relationship between sleep-disordered breathing and insulin resistance, indicated by fasting serum insulin level and insulin resistance index based on the homeostasis model assessment method (HOMA IR). A total of 270 consecutive subjects (197 male) who were referred for polysomnography and who did not have known diabetes mellitus were included, and 185 were documented to have OSA defined as an apnea-hypopnea index (AHI) > or =5. OSA subjects were more insulin resistant, as indicated by higher levels of fasting serum insulin (p = 0.001) and HOMA-IR (p < 0.001); they were also older and more obese. Stepwise multiple linear regression analysis showed that obesity was the major determinant of insulin resistance but sleep-disordered breathing parameters (AHI and minimum oxygen saturation) were also independent determinants of insulin resistance (fasting insulin: AHI, p = 0.02, minimum O(2), p = 0.041; HOMA-IR: AHI, p = 0.044, minimum O(2), p = 0.022); this association between OSA and insulin resistance was seen in both obese and nonobese subjects. Each additional apnea or hypopnea per sleep hour increased the fasting insulin level and HOMA-IR by about 0.5%. Further analysis of the relationship of insulin resistance and hypertension confirmed that insulin resistance was a significant factor for hypertension in this cohort. Our findings suggest that OSA is independently associated with insulin resistance, and its role in the atherogenic potential of sleep disordered breathing is worthy of further exploration. PMID- 11874813 TI - Sleep-disordered breathing and insulin resistance in middle-aged and overweight men. AB - Sleep-disordered breathing is a prevalent condition associated with impairment of daytime function and may predispose individuals to metabolic abnormalities independent of obesity. The primary objective of this study was to determine the metabolic consequences and community prevalence of sleep-disordered breathing in mildly obese, but otherwise healthy, individuals. One hundred and fifty healthy men, without diabetes or cardiopulmonary disease, were recruited from the community. Measurements included polysomnography, a multiple sleep latency test, an oral glucose tolerance test, determination of body fat by hydrodensitometry, and fasting insulin and lipids. The prevalence of sleep-disordered breathing, depending on the apnea-hypopnea index (AHI) cutoff, ranged from 40 to 60%. After adjusting for body mass index (BMI) and percent body fat, an AHI gt-or-equal, slanted 5 events/h was associated with an increased risk of having impaired or diabetic glucose tolerance (odds ratio, 2.15; 95% CI, 1.05-4.38). The impairment in glucose tolerance was related to the severity of oxygen desaturation (DeltaSa(O(2))) associated with sleep-disordered breathing. For a 4% decrease in oxygen saturation, the associated odds ratio for worsening glucose tolerance was 1.99 (95% CI, 1.11 to 3.56) after adjusting for percent body fat, BMI, and AHI. Multivariable linear regression analyses revealed that increasing AHI was associated with worsening insulin resistance independent of obesity. Thus, sleep disordered breathing is a prevalent condition in mildly obese men and is independently associated with glucose intolerance and insulin resistance. PMID- 11874814 TI - Chemical predictors of wheeze among farmer pesticide applicators in the Agricultural Health Study. AB - Pesticides may contribute to respiratory symptoms among farmers. Using the Agricultural Health Study, a large cohort of certified pesticide applicators in Iowa and North Carolina, we explored the association between wheeze and pesticide use in the past year. Self-administered questionnaires contained items on 40 currently used pesticides and pesticide application practices. A total of 20,468 applicators, ranging in age from 16 to 88 years, provided complete information; 19% reported wheezing in the past year. Logistic regression models controlling for age, state, smoking, and history of asthma or atopy were used to evaluate associations between individual pesticides and wheeze. Among pesticides suspected to contribute to wheeze, paraquat, three organophosphates (parathion, malathion, and chlorpyrifos), and one thiocarbamate (S-ethyl-dipropylthiocarbamate [EPTC] ) had elevated odds ratios (OR). Parathion had the highest OR (1.5, 95% confidence interval [CI] 1.0, 2.2). Chlorpyrifos, EPTC, paraquat, and parathion demonstrated significant dose-response trends. The herbicides, atrazine and alachlor, but not 2,4-D, were associated with wheeze. Atrazine had a significant dose-response trend with participants applying atrazine more than 20 days/year having an OR of 1.5 (95% CI 1.2,1.9). Inclusion of crops and animals into these models did not significantly alter the observed OR. These associations, though small, suggest an independent role for specific pesticides in respiratory symptoms of farmers. PMID- 11874815 TI - Polymorphisms in the tumor necrosis factor-alpha gene promoter may predispose to severe silicosis in black South African miners. AB - Susceptibility to silicosis is in part genetically determined. Polymorphisms in the promoter region of tumor necrosis factor (TNF)-alpha, a cytokine with a central role in the pathophysiology of silicosis, have been associated with predisposition to several infectious and inflammatory diseases. Polymorphisms at positions -308, -238, and -376 in the TNF-alpha promoter region were compared in nine patients with severe silicosis with International Labour Office (ILO) grade 3 nodularity, 112 patients with less severe silicosis (ILO grades 1/1 to 2/2), and 120 black South African gold miners without silicosis (ILO grades 0/0) in an age-frequency-matched case- control study. There were no significant differences between miners with less severe silicosis and controls at any loci in the TNF alpha promoter region, but miners with severe silicosis were significantly more likely than controls to have -238A (33% versus 6%, Fisher's exact p value = 0.022) and -376A (33% versus 5%, Fisher's exact p value = 0.016). These alleles were in linkage disequilibrium (p < 0.001), and so were not independent. The association remained significant (Fisher's exact p values = 0.011 and 0.011, respectively) when analysis was limited to the majority tribe (Basotho), which included all subjects with severe silicosis. Subjects with severe silicosis were also significantly more likely to have the -308A allele (Fisher's exact p value = 0.034), but this result was confounded by ethnicity and was not significant within Basotho tribe members (Fisher's exact p value = 0.15). TNF-alpha promoter polymorphisms are associated with severe, but not less severe, silicosis in this population. A predominant effect on disease severity, rather than on disease frequency, appears to be a general feature of promoter polymorphism in diseases in which TNF-alpha has a critical role. PMID- 11874816 TI - Internet-based home monitoring of pulmonary function after lung transplantation. AB - Home monitoring of spirometry has been advocated in lung transplant recipients for the early detection of acute infection and rejection of the allograft. We have developed a user-friendly, Internet-based telemonitoring system providing direct transmission of home spirometry to the hospital. In this prospective study, we assessed patient adherence with the monitoring, agreement between home and hospital spirometry, intrasubject coefficient of variation (CV) for FEV(1) and FEF(25-75), and sensitivity of these variables for the detection of acute complications. Twenty-two bilateral-lung and heart-lung transplant recipients were followed for a median of 473 d (range, 60-822), during which 13,833 measurements were obtained. Patient compliance was 55% for two measurements a day and 84% for one measurement a day. Agreement between home and hospital spirometry was within 4% for FEV(1) and 6% for FEF(25-75). Mean CV was 3.2% for FEV(1) and 7.5% for FEF(25-75). Using transbronchial lung biopsy and/or bronchoalveolar lavage as gold standards, the sensitivity of home spirometry was 63%, and 23% of true positives were detected by changes in FEF(25-75) alone. We conclude that home monitoring of pulmonary function in lung transplant recipients via the Internet is feasible and provides very reproducible data; yet it has only a mild sensitivity for the detection of acute allograft dysfunction. PMID- 11874817 TI - Comparison of nebulized budesonide and oral prednisolone with placebo in the treatment of acute exacerbations of chronic obstructive pulmonary disease: a randomized controlled trial. AB - Nebulized budesonide has been used successfully to treat acute asthma exacerbation, and we hypothesized that it could also be effective for exacerbations of chronic obstructive pulmonary disease (COPD). In this multicenter, double-blind, randomized, placebo-controlled trial, the efficacy of nebulized budesonide (Pulmicort Respules/Nebuamp), oral prednisolone, and placebo was compared in 199 patients with acute exacerbations of COPD requiring hospitalization. Patients received from randomization (H(0)) to 72 h (H(72)), 2 mg of budesonide every 6 h (n = 71), 30 mg of oral prednisolone every 12 h (n = 62), or placebo (n = 66). All received standard treatment, including nebulized beta(2)-agonists, ipratropium bromide, oral antibiotics, and supplemental oxygen. The mean change (95% confidence interval) in postbronchodilator FEV(1) from H(0) to H(72) was greater with active treatments than with placebo: budesonide versus placebo, 0.10 L (0.02 to 0.18 L); prednisolone versus placebo, 0.16 L (0.08 to 0.24 L). The difference in FEV(1) between budesonide and prednisolone was not significant, -0.06 L (-0.14 to 0.02 L). The occurrence of serious adverse events was similar for all groups. Budesonide had less systemic activity than prednisolone as indicated by a higher incidence of hyperglycemia observed with prednisolone. Both budesonide and prednisolone improved airflow in COPD patients with acute exacerbations when compared with placebo. Nebulized budesonide may be an alternative to oral prednisolone in the treatment of nonacidotic exacerbations of COPD but further studies should be done to evaluate its long-term impact on clinical outcomes after an initial episode of COPD exacerbation. PMID- 11874818 TI - The impact of chronic obstructive pulmonary disease on work loss in the United States. AB - Chronic obstructive pulmonary disease (COPD) is a rapidly growing public health problem in the United States and elsewhere. Although direct costs of COPD are well documented, the impact of COPD and its severity on labor force participation is not well known. Using population-based data from the Third National Health and Nutrition Examination Survey (NHANES III), we determined the adjusted relationship between COPD (and its severity) and labor force participation in the U.S. We used data from 12,436 participants involved in NHANES III; 1,073 of these participants (8.6% of the total) reported COPD. These participants were 3.9% (95% confidence interval, 1.3% to 6.4%) less likely to be in the labor force than those without COPD. Increasing severity of COPD was associated with decreased probability of being in the labor force (p for linear trend = 0.001). Mild, moderate, and severe COPD was associated with a 3.4%, 3.9%, and 14.4% reduction in the labor force participation rate relative to those without COPD. These data suggest that COPD has a considerable adverse impact on work force participation. Based on these data, we estimate that, in 1994, COPD was responsible for work loss of approximately $9.9 billion in the U.S. PMID- 11874819 TI - Role of serum cortisol levels in children with asthma. AB - Decreased serum cortisol levels have been proposed to contribute to nocturnal airway obstruction. We investigated whether endogenous cortisol levels are lower, and also whether the 24-h cortisol variation is greater, in children with asthma than in control subjects and assessed the relationship between serum cortisol and nocturnal airflow limitation in children with asthma. Cortisol and FEV(1) were measured every 4 h over 24 h; blood eosinophils, airway responsiveness to methacholine and adenosine 5'-monophosphate (AMP) were measured at 0400 and 1600. Children with asthma had lower cortisol levels than did control subjects; at midnight the difference was significant. Subjects with nocturnal asthma (24-h FEV(1) variation > or =15%) had significantly lower cortisol levels than did control subjects at 0000, 0800, and 1200. A higher mean 24-h cortisol level in subjects with asthma was associated with a significantly higher FEV(1) as a percentage of the predicted value (FEV(1) %pred) at 0400, 0800, and 2000, yet not in control subjects. Higher 24-h cortisol variation was associated with lower FEV(1) %pred at all time points in both control subjects and subjects with nonnocturnal asthma. There was no significant association between the level or variation of cortisol and PD(20) methacholine (provocative dose of methacholine causing a 20% fall in FEV(1)), PD(20) AMP, or eosinophils. Our data suggest that lower cortisol levels contribute to both overall lower levels of FEV(1) especially at night. This may be due to a lack of suppression of airway inflammation. PMID- 11874820 TI - Pneumococcal meningitis in the intensive care unit: prognostic factors of clinical outcome in a series of 80 cases. AB - We have undertaken this retrospective study to determine factors associated with in-hospital mortality and morbidity in 80 adult patients with severe Streptococcus pneumoniae meningitis. Clinical characteristics at admission of patients infected with susceptible (n = 54) and nonsusceptible (n = 17) strains to penicillin G were similar: age: 51 +/- 19 versus 58 +/- 15 yr (p = 0.16); Simplified Acute Severity Score (SAPS II): 39 +/- 14 versus 41 +/- 11 (p = 0.68); and Glasgow Coma Score: 8 +/- 3 versus 9.5 +/- 3 (p = 0.21), respectively. In hospital mortality was 25% (20/80), with one death among the 17 patients (6%) infected with a nonsusceptible strain (p = 0.03). High-dose dexamethasone was used in 22 cases. By multivariate analysis, three factors were independently associated with death: platelet count < 100 G/L (adjusted odds ratio [aOR] = 32.7; 95% CI = 3.2 to 332.5; p = 0.0032), arterial pH > 7.47 (aOR = 33.1; 95% CI = 3.4 to 319.7; p = 0.0025), and mechanical ventilation (aOR = 48.8; 95% CI = 2.6 to 901.5; p = 0.009). When adjusting for the identified prognostic factors, corticosteroids significantly reduced the risk of death (aOR = 0.069; 95% CI = 0.005 to 0.9; p = 0.048). Only SAPS II was predictive of adverse outcome (death or neurologic deficit). We conclude that in intubated patients with S. pneumoniae meningitis, hyperventilation should be used with caution. Nonsusceptibility to penicillin G is not associated with a worse outcome. High-dose corticosteroids may be beneficial in the most severely ill patients. PMID- 11874821 TI - A pilot study of all-trans-retinoic acid for the treatment of human emphysema. AB - Emphysema results from progressive destruction of alveolar septae and was considered irreversible until all-trans-retinoic acid (ATRA) was shown to reverse anatomic and physiologic signs of emphysema in a rat model. To evaluate the feasibility of ATRA as a clinical therapy, 20 patients with severe emphysema were enrolled into a randomized, double-blind, placebo-controlled pilot study. Participants included 16 male and 4 female former smokers, two with alpha(1) antitrypsin deficiency. Patients were treated with either 3 mo of ATRA (50 mg/m(2)/d) or 3 mo of placebo, followed by a 3-mo crossover phase. Plasma drug levels were followed and outcome measures included serial pulmonary function tests, blood gases, lung compliance, computed tomography (CT) imaging, and quality of life questionnaires. In general, treatment was well tolerated and associated with only mild side effects including skin changes, transient headache, hyperlipidemia, transaminites, and musculoskeletal pains. Plasma drug levels varied considerably between subjects and decreased significantly over time in 35% of the participants. Physiologic and CT measurements did not change appreciably in response to therapy. We conclude that ATRA is well tolerated in patients with emphysema, and trials evaluating higher doses, longer treatment, or different dosing schedules are feasible. PMID- 11874822 TI - Blockade of CD28/B7-2 costimulation inhibits experimental obliterative bronchiolitis in rat tracheal allografts: suppression of helper T cell type1 dominated immune response. AB - T cell activation is a proximal event in the initiation of chronic rejection that may ultimately lead to obliterative bronchiolitis (OB) after lung transplantation. In addition to primary signals generated by the T cell receptor, T cell activation relies on costimulatory signals, of which the most important are mediated via interaction between CD28 and its ligands B7-1 and B7-2. In nontreated rat tracheal allografts, B7-2, but not B7-1, expression peaked 10 d after transplantation, unlike in syngeneic grafts, where no B7-2 upregulation was observed. Selective blockade of the CD28/B7-1 T cell costimulatory pathway by a mutant form of CTLA4Ig (cytotoxic T lymphocyte antigen 4 immunoglobulin), CTLA4IgY100F, did not affect epithelial injury or degree of luminal occlusion in rat tracheal allografts. Treatment with CTLA4Ig fusion protein, which blocks both CD28/B7-1 and CD28/B7-2 interaction, significantly delayed the development of epithelial injury and airway occlusion. Immunohistochemical analyses of allografts showed that selective inhibition of the CD28/B7-1 pathway did not affect cytokine expression. In contrast, treatment with CTLA4Ig was associated with a significant decrease in the intragraft production of tumor necrosis factor alpha, interleukin 2, and interferon gamma, as well as slightly increased intragraft expression of interleukin 10. In conclusion, CTLA4Ig-mediated costimulatory blockade delays epithelial injury and attenuates obliterative changes and is associated with marked suppression of helper T cell type 1 (Th1) dominated cytokine response. These observations emphasize the role of the CD28/B7 2 costimulatory pathway in regulating proinflammatory and Th1 cytokine responses and thereby in the development of epithelial and graft injury gradually leading to obliteration of the airway lumen. PMID- 11874824 TI - PC(20) adenosine 5'-monophosphate is more closely associated with airway inflammation in asthma than PC(20) methacholine. PMID- 11874825 TI - More inflammation than lung in emphysema. PMID- 11874826 TI - The discriminatory capacity of the bronchodilator response. PMID- 11874827 TI - Newly diagnosed rheumatoid arthritis. PMID- 11874830 TI - Polymyositis as a cause of total gut failure. AB - BACKGROUND: Gastrointestinal manifestations are seen in systemic sclerosis and mixed connective tissue disorders but are rare in pure polymyositis. CASE REPORT: A 44 year old woman with polymyositis who developed total gut failure requiring treatment with total parenteral nutrition is described. RESULTS: The patient's polymyositis is now fully controlled biochemically, but her gastrointestinal symptoms persist. PMID- 11874829 TI - Tumour necrosis factor (TNF) in psoriatic arthritis: pathophysiology and treatment with TNF inhibitors. AB - High levels of proinflammatory cytokines, including tumour necrosis factor (TNF), have been detected in psoriatic skin lesions and joints of patients with the inflammatory disease. Early results of treatment of psoriatic arthritis and psoriasis with TNF neutralising agents are encouraging, but whether these agents will be able to improve long term outcomes, such as disability, is not yet known. PMID- 11874828 TI - Early referral recommendation for newly diagnosed rheumatoid arthritis: evidence based development of a clinical guide. AB - BACKGROUND: Effective treatment of active rheumatoid arthritis (RA) requires early diagnosis and early disease modifying antirheumatic drug (DMARD) treatment to have an impact on long term morbidity and mortality. Clinical criteria would facilitate early referral of the patient with suspected RA to a rheumatologist for definitive diagnosis and initiation of DMARD treatment at that point in the disease most likely to have an impact on the long term outcome. OBJECTIVE: To develop a referral recommendation that may serve as a clinical guide for primary care doctors, enabling them to identify patients with suspected RA during the early inflammatory stages. METHODS: Key points of the referral criteria were formed based on a thorough literature review targeting early RA, early arthritis clinics, DMARD treatment for early RA, prognostic factors of disease progression, early RA clinical trials, and quality of life. Evidence was graded using the methods defined by Shekelle et al. A draft version of the criterion was circulated among the authors for critical evaluation. A consensus integrated these comments. RESULTS: Clinical evidence strongly supports the observations that structural damage occurs early in active RA and that early DMARD treatment improves the long term outcome of the disease. The observations indicate that rapid referral to a rheumatologist is advised when RA is suspected. This may be supported by the presence of any of the following: >or=3 swollen joints, metatarsophalangeal/metacarpophalangeal involvement, and morning stiffness of >or=30 minutes. CONCLUSION: The proposed early referral recommendation is a viable tool for primary care doctors to identify potential patients with active RA early in the disease. Early referral to a rheumatologist for definitive diagnosis and early DMARD treatment should improve the long term outcome of RA. PMID- 11874831 TI - Effect of pregnancy and obstructive jaundice on inflammatory diseases: the work of P S Hench revisited. AB - Hench considered that cortisone improved inflammatory joint symptoms during pregnancy and obstructive jaundice. However, the improved symptoms are probably due to changes in the proportions of fatty acids in plasma and inflammatory cell phospholipids. These changes decrease the superoxide anions and eicosanoids produced and also reduce tumour necrosis factor alpha production. PMID- 11874834 TI - Characterisation of size and direction of osteophyte in knee osteoarthritis: a radiographic study. AB - OBJECTIVES: To examine the size and direction of osteophyte in knee osteoarthritis (OA) and to determine associations between osteophyte size and other radiographic features. METHODS: Knee radiographs (standing extended anteroposterior and 30 degrees flexion skyline views) were examined from 204 patients referred to hospital with symptomatic knee OA (155 women, 49 men; mean age 70, range 34-91 years). A single observer assessed films for osteophyte size and direction at eight sites; narrowing in each compartment; varus/valgus angulation; patellofemoral subluxation; attrition; and chondrocalcinosis using a standard atlas, direct measurement, or visual assessment. For analysis, one OA knee was selected at random from each subject. RESULTS: Osteophyte direction at the eight sites was divisible into five categories. At all sites, except for the lateral tibial plateau and the medial patella, osteophyte direction varied according to (a) the size of osteophyte and (b) the degree of local narrowing. At the medial femur, medial tibia, and lateral femur osteophyte direction changed from being predominantly horizontal to predominantly vertical with increasing size. The size of osteophyte correlated positively with the severity of local narrowing, except for the medial patellofemoral compartment where osteophyte size correlated positively with the severity of narrowing in the medial tibiofemoral compartment. Logistic regression analysis showed that osteophyte size was associated not only with local narrowing but also with local malalignment and bone attrition, and that chondrocalcinosis was positively associated with osteophyte size at multiple sites. CONCLUSION: In patients referred to hospital with knee OA different patterns of osteophyte direction are discernible. Osteophyte size is associated with local compartmental narrowing but also local alignment and attrition. Chondrocalcinosis is associated with osteophytosis throughout the joint. These data suggest that both local biomechanical and constitutional factors influence the size and direction of osteophyte formation in knee OA. PMID- 11874832 TI - Long term anti-tumour necrosis factor alpha monotherapy in rheumatoid arthritis: effect on radiological course and prognostic value of markers of cartilage turnover and endothelial activation. AB - OBJECTIVES: To investigate the effect of prolonged neutralisation of tumour necrosis factor alpha (TNFalpha) on the radiological course in rheumatoid arthritis (RA). To assess whether the radiological course can be predicted by clinical variables or biological markers of cartilage and synovium turnover and of endothelial activation. PATIENTS AND METHODS: Forty seven patients with active RA enrolled at our centre in monotherapy trials with adalimumab (D2E7), a fully human anti-TNFalpha monoclonal antibody, were studied for two years. Radiographs of hands and feet obtained at baseline and after one and two years were scored in chronological order by a single, blinded observer using the modified Sharp method. Radiological course was classified as stable or progressive using the smallest detectable difference as cut off point. The relation between radiological course and serum markers of cartilage and synovium turnover (metalloproteinases (MMP-1 and MMP-3), cartilage oligomeric matrix protein (COMP), human cartilage glycoprotein-39 (HC gp-39)), endothelial activation (soluble E-selectin and intercellular adhesion molecule (ICAM-1)), and integrated measures of disease activity were assessed using univariate and multivariate analysis. RESULTS: Radiological evaluation was performed in 36 patients with paired sets of radiographs at baseline and two years. After two years a total of 15/36 (42%) presented no radiological progression. More patients with stable radiological course were still receiving anti-TNFalpha treatment after two years (13/15 (87%) v 11/21 (52%); p=0.03) and had lower baseline COMP and sICAM-1 levels (p=0.01 and 0.04, respectively) than those in the group with progressive disease. In a logistic regression model the combination of sustained TNF neutralisation and baseline COMP and sICAM-1 levels was predictive for radiological outcome (p=0.03). C reactive protein and disease activity score area under the curve were significantly correlated with changes in radiological scores after two years (r=0.40 and 0.37, p<0.05). Long term TNFalpha neutralisation decreased the levels of COMP, sICAM, MMPs, and HC gp-39, but not sE-selectin. CONCLUSION: The results suggest that long term monotherapy with anti-TNFalpha has a positive effect on radiological outcome and modulates cartilage and synovium turnover as measured by biological markers. Baseline serum sICAM-1 levels and COMP levels may be helpful to identify patients with progressive or non progressive radiological outcome. PMID- 11874835 TI - Bone quality and bone mass as assessed by quantitative ultrasound and dual energy x ray absorptiometry in women with rheumatoid arthritis: relationship with quadriceps strength. AB - OBJECTIVE: To examine relationships of bone quality as assessed by quantitative ultrasound (QUS) and bone mineral density (BMD, g/cm(2)) with quadriceps strength (QS) in women with rheumatoid arthritis (RA). METHODS: Sixty seven women with RA according to the 1987 American College of Rheumatology (ACR) criteria were examined. Mean (SD) age was 62 (13) years, mean disease duration 15 years. Most were or had been receiving glucocorticoid treatment. Calcaneal bone quality expressed as speed of sound (SOS, m/s), broadband ultrasound attenuation (BUA, dB/MHz), and stiffness was measured by QUS. BMD of the femoral neck, spine, and distal forearm was measured by dual energy x ray absorptiometry (DXA). Maximal voluntary isokinetic quadriceps strength (Nm) was assessed by isokinetic dynamometry. Pain was recorded on a visual analogue scale (VAS), disability was scored by the Stanford Health Assessment Questionnaire (HAQ), and the degree of physical impairment was expressed by the Steinbrocker index (SI). RESULTS: In multiple regression analyses, QS predicted SOS, BUA, and stiffness (r(partial) ranging from 0.36 to 0.45, p<0.005) and femoral neck BMD (r(partial)=0.30, p<0.05) independently of age, height, weight, disease duration, HAQ, VAS, SI, and cumulative steroid dose. BMD of the spine and distal forearm was not associated with QS. After adjustment for covariates, women with subnormal BMD of the femoral neck (T score <-1), had a 20% lower QS than those with normal BMD (p<0.0001). CONCLUSIONS: Calcaneal bone quality and femoral neck BMD were associated with QS in women with RA. This finding indicates that physical activity including muscle strengthening exercises may play a part in the prevention of bone loss in these patients. PMID- 11874836 TI - Development of a radiographic index to assess the tarsal involvement in patients with spondyloarthropathies. AB - OBJECTIVE: To develop and test an index to evaluate the radiographic changes that occur in the tarsus and adjacent areas of the foot in patients with spondyloarthropathies (SpA). METHODS: The spondyloarthropathy tarsal radiographic index (SpA-TRI) was developed in three consecutive steps: (a) detection of descriptors after reviewing 70 radiographic files; (b) descriptor gradation and subsequent modifications performed by a consensus committee, and (c) interobserver variability assessed by three blinded and independent observers on 272 radiographs: anteroposterior 118, lateral 90, oblique 64 from 121 patients with SpA, and intraobserver variability on 75 radiographs from 25 patients with SpA. Statistical analysis included percentage of agreement and kappa test. SpA TRI score ranges from 0 to 4 (0=normal; 1=osteopenia or suspicious findings; 2=definite joint space narrowing, bony erosion(s), periosteal whiskering, or enthesophyte(s) in the plantar fascia or Achilleal tendon attachments; 3=para articular enthesophyte(s); 4=bony ankylosis (joint space fusion or complete bridging)). RESULTS: Complete agreement for every evaluation was >40%, and discordance >1 grade was <15%. The kappa scores among the three observers were acceptable for all the single projections: oblique (0.52, 0.36, 0.35), lateral (0.50, 0.42, 0.56), and anteroposterior (0.40, 0.41, 0.21) views. The combination of lateral and oblique views achieved the highest concordance rates (0.72, 0.33, 0.66), surpassing that of the three projections altogether (0.34, 0.58, 0.37). In every case the concordance was comparable with that of sacroiliac joints (0.47, 0.41, 0.34); intraobserver concordance showed a similar trend. CONCLUSION: The SpA-TRI is an index that includes the most prominent features of tarsal disease and adjacent areas of the foot in SpA and grades them accordingly, it has an adequate reproducibility, and is suitable for use with two or more projections, preferably the combination of oblique and lateral. PMID- 11874837 TI - Which patients stop working because of rheumatoid arthritis? Results of five years' follow up in 732 patients from the Early RA Study (ERAS). AB - OBJECTIVES: To assess the occurrence and prognostic factors for the ability to maintain paid work in patients with rheumatoid arthritis (RA). SETTING: Inception cohort of patients with RA recruited from rheumatology departments in nine NHS Hospital Trusts in England. PATIENTS: All consecutive patients with RA of less than two years' duration, before any second line (disease modifying) drug treatment, and followed up for five years. METHODS: Clinical, laboratory, and radiological assessments, and all treatments were recorded prospectively using a standardised format at presentation and yearly. OUTCOME MEASURES: Changes in, and loss of paid work by five years' follow up. RESULTS: 732 patients completed the five year follow up. 353/721 (49%) were gainfully employed at the onset of RA, 211 (60%) were still working at five years, 104 (29%) stopped because of the disease, and 31 (9%) retired for reasons other than RA. Work disability at five years was more likely in manual workers (odds ratio (OR) 2.3, 95% confidence interval (CI) 1.4 to 3.8) and worse baseline Health Assessment Questionnaire (HAQ>1.5, OR 2.26, 95% CI 1.38 to 3.7). In combination with other baseline variables (erythrocyte sedimentation rate, sex, age of onset, and radiological erosions), employment outcome was predicted in 78% using multivariate analysis. CONCLUSIONS: Nearly half of the patients with RA were in paid employment at onset, work disability was an adverse outcome for a third of these patients by five years, and manual work and high baseline HAQ were important predictors for this. These details are likely to be useful to clinicians, health professionals, and patients in order to plan medical, orthopaedic, and remedial treatments in early RA. Future disease modifying treatments could be compared with this cohort of patients who were treated with conventional second line drugs. PMID- 11874838 TI - Review of the function of a telephone helpline in the treatment of outpatients with rheumatoid arthritis. AB - OBJECTIVES: To examine the role, acceptability, and cost effectiveness of a telephone helpline organised and run by specialist nurses in a district general hospital outpatient rheumatology department. MATERIAL AND METHODS: Patients accessed the telephone helpline by leaving a taped message on an answer phone with a 24 hour response time. Assessment included an audit of the nature and outcome of helpline calls, patient satisfaction with the helpline, and a health economic analysis of the helpline operation. A postal questionnaire was used to assess patient satisfaction; this was sent to the 87 patients who called the helpline during one month, and overall satisfaction with the helpline was assessed. The nature and outcome of all calls was analysed retrospectively using a helpline record book for February and October of one year and February of the following year. From the results of the retrospective analysis and an estimate of the number of general practitioner consultations avoided by provision of the helpline, the cost effectiveness of the helpline was calculated. RESULTS: Of those returning questionnaires, 61/63 (97%) were satisfied with the response time, 63/63 (100%) with the courtesy, and 60/63 (95%) felt that their questions were answered directly and to their satisfaction in 62 (98%) of cases. Had the helpline not been available, 38/63 (60%) patients would have made an appointment with their GP. When these figures were extrapolated to an annual estimation, a basic cost analysis showed that the helpline produced a cost saving to the NHS, largely as a result of GP consultations avoided. CONCLUSION: Clinical advice and support can be provided by a rheumatology helpline set up as an adjunct to a standard outpatient service. The results of a postal questionnaire suggested more than 95% satisfaction with all aspects of the helpline service and that 99% of callers would call the helpline again. The provision of the helpline service contributes to the quality of care provided by an outpatient department and provides benefit to the NHS. PMID- 11874840 TI - Influence of HLA polymorphism on persistent remission in rheumatoid arthritis. AB - BACKGROUND: Several studies have reported an association between the presence of the shared epitope (SE) and susceptibility to rheumatoid arthritis (RA). Recent studies have shown that certain HLA-DRB1 alleles in combination with predisposing DQB1 and DQA1 alleles may protect against the development of RA. This model is known as the rheumatoid arthritis protection (RAP) hypothesis. OBJECTIVE: To determine the distribution of HLA-DRB1 and DQB1/DQA1 alleles in a cohort of patients with RA in remission and to determine the association between these HLA alleles and the persistence of remission. PATIENTS AND METHODS: HLA-DRB1 and DQB1 typings were performed in 167 patients with RA in remission, defined according to the American College of Rheumatology criteria. The disease course, as defined by the persistence of remission during a follow up of two years, was compared between subgroups. According to the RAP hypothesis patients were divided into three subgroups: patients carrying predisposing DQ alleles, patients carrying predisposing alleles in combination with protective alleles (DQ(RA+)/DERAA phenotype), and patients lacking the predisposing alleles. According to the SE hypothesis, patients were divided into three subgroups based on whether they were carrying two, one, or no predisposing alleles (SE alleles). RESULTS: Predisposing DQ alleles along with a DERAA-bearing allele were present in 14 (8%) of the 167 patients. At least one SE allele was present in 116 (69%) patients; 34 of them (20%) were carrying two copies. The disease course was not significantly different between the subgroups according to the SE and RAP hypothesis, respectively. CONCLUSION: The frequency of DQ(RA+)/DERAA combinations and of SE alleles in patients with RA clinically in remission was similar to that found in other RA populations. Persistent remission of RA was not associated with any particular HLA subtypes, indicating that HLA typing is not useful for predicting persistent clinical remission. PMID- 11874839 TI - Influence of prior pregnancies on disease course and cause of death in systemic sclerosis. AB - BACKGROUND: Microchimerism from fetal or maternal cells transferred during pregnancy has been implicated in the pathogenesis of systemic sclerosis (SSc). OBJECTIVE: To determine whether a prior pregnancy influenced disease progression and cause of death in patients with SSc. PATIENTS AND METHODS: The patients comprised a retrospective study cohort of 111 women with SSc: 78 patients with prior pregnancies (PP) and 33 who were never pregnant (NP), followed up at Thomas Jefferson University. Differences in age at onset, disease subset, organ involvement, cause of death, and type of antinuclear autoantibodies were evaluated statistically, including regression analysis. RESULTS: The age at onset of SSc in NP patients was 32.0 years compared with 45.7 years in patients with one or two prior pregnancies (p<0.0001), 46.6 years in patients with three or four pregnancies (p<0.0001), and 51.3 years in patients with five to seven pregnancies (p<0.0005). In the 16 patients who had an elective pregnancy termination, 14/16 (87.5%) had diffuse SSc v 2/16 (12.5%) with limited SSc (p<0.0001; odds ratio (OR)=49.0). Of the NP women, 7/30 (23%) died from SSc related causes v 3/78 (4%) women who had pregnancies (p=0.0058; OR=7.6). A carbon monoxide transfer factor (TLCO) of <60% and disease duration >10 years was found in 10/13 (77%) NP patients v 10/23 (43%) patients who had pregnancies (p=0.05; OR=4.7), and a TLCO <50% and disease duration >10 years was identified in 7/13 (54%) NP patients v 6/23 (26%) of the patients who had pregnancies (p=0.09; OR=3.2). CONCLUSIONS: There are differences in the age at onset, clinical course, severity of lung involvement, and cause of death in women who develop SSc before pregnancy compared with those who develop it after pregnancies. The NP patients with SSc had onset of disease at an earlier age, more severe lung involvement, and higher rate of death due to SSc. PMID- 11874841 TI - Juvenile idiopathic arthritis (JIA) is primarily associated with HLA-DR8 but not DQ4 on the DR8-DQ4 haplotype. AB - BACKGROUND: Juvenile idiopathic arthritis (JIA) is strongly associated with the DR8-DQ4 haplotype. The genes encoding DR8 and DQ4 are in strong linkage disequilibrium (LD) and occur together on the same HLA haplotype in almost all patients and controls. Because of the strong LD it is not clear whether DR8, DQ4, or both, are primarily associated with JIA. OBJECTIVE: To unveil the primary association of JIA--that is, with DR8 or DQ4. METHODS: DRB1, DQA1, and DQB1 alleles of 585 Norwegian and 47 Polish unrelated patients with JIA (categorised as pauciarticular and rheumatoid factor negative polyarticular JIA), and of 3155 Norwegian and 158 Polish unrelated controls, were typed using a polymerase chain reaction or oligonucleotide hybridisation and sequence-specific primers method. RESULTS: Several haplotypes which encoded DR8 (that is, carried DRB1*08) and which did not encode DQ4 (that is, did not carry DQA1*0401) were found. Such haplotypes were found in three Norwegian patients and two controls (p=0.029). In the Polish population such haplotypes were found among four patients with JIA and two controls (p=0.025). No haplotypes which carried DQA1*0401 and DQB1*0402 in the absence of DRB1*08 were found, either among patients with JIA (Polish and Norwegian) or among the controls (Polish). CONCLUSION: On the DR8-DQ4 haplotype the DRB1*08 allele is primarily associated with JIA. PMID- 11874843 TI - Anticardiolipin antibodies in rheumatoid patients treated with etanercept or conventional combination therapy: direct and indirect evidence for a possible association with infections. AB - OBJECTIVE: To assess the occurrence of anticardiolipin antibodies (ACA) (as well as of anti-DNA antibodies) in patients with rheumatoid arthritis treated with etanercept or combination therapy. METHODS: Eight patients treated with etanercept 25 mg twice weekly were studied for a period of 85 weeks. A control group of 39 patients with rheumatoid arthritis undergoing combination treatment (methotrexate (MTX) + cyclosporin A or MTX + chloroquine) were studied for the same period of time. The occurrence of anticardiolipin antibodies (ACA-IgG) and anti-DNA was examined, together with the possible occurrence of infections due to bacteria capable of inducing B cell activation. RESULTS: In 5/8 patients receiving etanercept an increase of ACA-IgG was seen, while anti-DNA became positive in 3/8 patients. A nasal or bronchial infection due to Staphylococcus aureus (Staph aureus) or a urinary tract infection due to E coli, occurred in all five cases. Antibiotic treatment produced a return to normal of ACA-IgG, and also of anti-DNA, in all cases except one. The infectious agent was eradicated in all subjects but one. In the control group Staph aureus was found in the nasal swab in 10/39 subjects; ACA-IgM (followed by ACA-IgG) appeared at the same time as infection occurred in 6/10, while no infection related to the increased ACA-IgM was recorded in the other four. CONCLUSIONS: Bacterial DNA, especially that enriched in CpG motifs, is a powerful immunostimulant that may, in some cases, lead to ACA or anti-DNA positivity, once tumour necrosis factor alpha is blocked. Eradication of the infections leads to a rapid decrease of ACA-IgG and of anti DNA levels. PMID- 11874844 TI - Limited polyarteritis nodosa of the male and female reproductive systems: diagnostic and therapeutic approach. AB - BACKGROUND: Polyarteritis nodosa (PAN) is a multisystem necrotising small and medium sized vasculitis that when left untreated carries a grave prognosis, with a five year survival of 10-15%. Prolonged immunosuppressive treatment with cyclophosphamide and steroids leads to high remission rates while carrying the risk of life threatening complications. The diagnostic and therapeutic approach for patients with isolated genital tract PAN is not well defined. OBJECTIVE: To present the management and follow up of two patients with limited PAN localised to the male and female reproductive system. CASE REPORTS: A 26 year old man presented with an "acute scrotum". He was afebrile and had no other sign or symptom. Laboratory tests, including complete blood count, erythrocyte sedimentation rate, liver and renal function tests, C reactive protein, antinuclear antibody, cryoglobulins, complement levels, antineutrophil cytoplasmic antibodies, and hepatitis B surface antigen, were all normal. His left testis was excised. Histopathology disclosed PAN of medium sized arteries with testicular infarction but no signs of torsion or infection. The other patient was a 51 year old woman who had had a total hysterectomy for a uterine myoma; incidentally PAN of the uterus and fallopian tubes was discovered. Neither patient received any immunosuppressive treatment after surgical removal of the affected organ. On prolonged follow up (clinical and laboratory evaluation) both patients are healthy with no sign of local recurrence or systemic PAN. PMID- 11874845 TI - Prevalence of Behcet's disease in an Arab community in Israel. AB - OBJECTIVE: To evaluate the prevalence of Behcet's disease (BD) in an Israeli Arab town (Taibe). METHODS: Questionnaires about the occurrence and prevalence of aphthous ulcers were distributed randomly to the parents of children attending a paediatric centre in Taibe. The parents were asked whether they or any of their children aged between 10 and 20 years had recurrent aphthous stomatitis. Any who had had more than four aphthous episodes (each episode lasting more than seven days) during the previous year were invited for an extensive interview and examination by a rheumatologist or a paediatrician. RESULTS: A total of 4876 subjects were included in this survey, of whom six (one male, five female) were diagnosed as having BD. Of these six, two were siblings (a brother and a sister). Five had skin lesions, four had visual involvement, and all had genital ulcers and joint symptoms; one in two patients had a positive pathergy test. Five of the six carried HLA-B5 antigens. The results showed a prevalence of 12/10,000 in Taibe. CONCLUSION: The prevalence of BD found in our survey is high and concurs with that found in other Mediterranean and Asian countries. PMID- 11874846 TI - Rapid detection of autoantibodies to dsDNA with the particle gel immunoassay (ID PaGIA). AB - OBJECTIVE: To describe a new particle agglutination test for the detection of autoantibodies to double stranded DNA (dsDNA). PATIENTS AND METHODS: Serum samples were collected from 40 unselected healthy blood donors and 200 patients with systemic lupus erythematosus (SLE) or a positive antinuclear antibody screen, or both. The samples were tested in the presence of red high density polystyrene particles coated with purified human dsDNA using the gel technique (Micro Typing System, ID-PaGIA, particle gel immunoassay). The results were compared with those obtained by the two standard anti-dsDNA antibody detection methods, Crithidia luciliae immunofluorescence test (CLIF) and enzyme linked immunosorbent assay (ELISA). RESULTS: The three anti-dsDNA assays exhibited an overall agreement of 87% and significant correlation with each other (p<0.0001). In the SLE group (n=71), 45 patients (63%) were found to be positive by ID-PaGIA compared with only 11/129 (9%) patients in the non-SLE group. Thus the ID-PaGIA had a sensitivity of 63%, and a specificity of 92% for SLE. In comparison, the standard detection methods showed sensitivities of 62% (CLIF) and 70% (ELISA) and specificities of 99% (CLIF) and 84% (ELISA) for SLE. Anti-dsDNA reactivity in the agglutination assay correlated closely with the quantities of antibody obtained by CLIF (r=0.81, p<0.0001) and ELISA (r=0.73, p<0.0001). CONCLUSIONS: The new particle gel agglutination test is a sensitive and specific immunoassay. It is a simple test procedure that might be well suited as a rapid screening method. PMID- 11874848 TI - Anticardiolipin antibodies in patients with post-streptococcal reactive arthritis. PMID- 11874847 TI - Progress towards an OMERACT-ILAR guideline for economic evaluations in rheumatology. AB - A working report from the OMERACT Health Economics Group. The group is working towards creating common standards for economic evaluation in rheumatology and also towards improving the scientific underpinning of economic evaluation, particularly pertaining to the rheumatic diseases. Preliminary recommendations for "reference cases" in osteoporosis, rheumatoid arthritis, and osteoarthritis are proposed. PMID- 11874849 TI - Successful treatment of SAPHO syndrome with infliximab: report of two cases. PMID- 11874850 TI - Muscle involvement in childhood sarcoidosis and need for muscle biopsy. PMID- 11874851 TI - Fever of unknown origin with seronegative spondyloarthropathy: an atypical manifestation of Whipple's disease. PMID- 11874852 TI - Synovial fluid neutrophil function in RA: the effect of pregnancy associated proteins. PMID- 11874853 TI - Behcet's disease and thrombophilia. PMID- 11874855 TI - Development and use of multimeric major histocompatibility complex molecules. PMID- 11874854 TI - Role of infectious and immune factors in coronary and cerebrovascular arteriosclerosis. PMID- 11874856 TI - Increased levels of inflammatory mediators in children and adults infected with Vibrio cholerae O1 and O139. AB - Investigations were carried out to study the production of factors associated with the innate immune response in the systemic and mucosal compartments in adults and children infected with Vibrio cholerae O1 and V. cholerae O139. The levels of nonspecific mediators of the innate defense system, i.e., prostaglandin E(2) (PGE(2)), leukotriene B(4) (LTB(4)), and lactoferrin (Lf), as well as myeloperoxidase (MPO), were elevated at the acute stage of the disease in stools obtained from both O1- and O139-infected adults and children. In the systemic compartment, the levels of Lf were increased after onset of disease, which in children remained elevated up to convalescence compared to the healthy controls. Increased concentrations of C-reactive protein were seen in the sera of adult cholera patients at the acute stage of infection. Elevated levels of the nitric oxide (NO*) metabolites (nitrite and nitrate [NO(2)(-) and NO(3)(-)]) were detected in plasma but not in urine. The activity of the scavenger of reactive oxygen species, superoxide dismutase, was higher in the plasma of adults immediately after the onset of disease, suggesting that an active scavenging of reactive oxygen species was taking place. The concentration of 8-iso prostaglandin F(2 alpha) remained unchanged in the systemic and mucosal compartments in the study subjects. After the recovery of patients from cholera, the concentration of the majority of the metabolites decreased to baseline levels by day 30 after the onset of infection. Immunohistochemical staining showed increased tissue expression of MPO, Lf, and inducible nitric oxide synthase at the acute stage in the duodenal biopsies of adults and rectal biopsies obtained from children with cholera. Very little difference was seen in the levels of the different inflammatory mediators in patients infected with V. cholerae O1 or the encapsulated V. cholerae O139. In summary, these results suggest that elevated concentrations of Lf, MPO, PGE(2), LTB(4), and NO*, as well as other metabolites, during the acute stage of the disease indicate that the innate defense system, as well as the inflammatory process, is activated in both adults and pediatric patients infected with V. cholerae O1 and O139. PMID- 11874857 TI - Depletion of human NK and CD8 cells prior to in vitro H1N1 flu vaccine stimulation increases the number of gamma interferon-secreting cells compared to the initial undepleted population in an ELISPOT assay. AB - In order to study the respective roles of CD4, CD8, and CD56 (NK) cells in gamma interferon (IFN-gamma) production after in vitro stimulation with flu vaccine in a healthy adult human population, we depleted these cellular subtypes before stimulation with antigen (inactivated split vaccine, A/Texas H1N1, or A/Sydney H3N2). We observed that while CD4 cells were required for IFN-gamma secretion in both conditions in vitro, CD56 (NK) cells and, to a lesser extent, CD8 cells had a negative effect on such synthesis upon H1N1 stimulation, as judged by an increased number of spots compared to the initial undepleted population. This regulation of IFN-gamma secretion was associated with an increase in ICAM-1 expression, in particular on T and B cells. This study points out the importance of evaluating in vitro immune responses on a whole-cell population in addition to isolated subtypes if one needs to address potential cellular interactions occurring in vivo in some situations (H1N1 stimulation in the present case). Such cross-regulations occur even in vitro during the antigenic stimulation step. PMID- 11874858 TI - Changes in CD4(+), CD8(+), CD4(+) CD8(+), and immunoglobulin M-positive peripheral blood mononuclear cells of postweaning multisystemic wasting syndrome affected pigs and age-matched uninfected wasted and healthy pigs correlate with lesions and porcine circovirus type 2 load in lymphoid tissues. AB - Forty-one 8- to 12-week-old wasted pigs were selected from several conventional farms with histories of postweaning multisystemic wasting syndrome (PMWS) and classified into two groups according to their porcine circovirus type 2 (PCV2) infection status, as determined by in situ hybridization (ISH). Twenty-four pigs tested positive for PCV2 (PCV2-positive group), while 17 pigs tested negative for PCV2 (PCV2-negative group). In addition, eight uninfected healthy pigs from an experimental farm were used as controls. Heparinized blood samples were taken to obtain peripheral blood mononuclear cells. The CD4(+), CD8(+), CD4(+) CD8(+) (double-positive [DP]), and immunoglobulin M-positive (IgM(+)) cell subsets were analyzed by flow cytometry with appropriate monoclonal antibodies. Histopathological studies were done to evaluate the apparent degrees of lymphocyte depletion in different lymphoid organs (superficial inguinal and mesenteric lymph nodes, Peyer's patches, tonsils, and spleen) and to determine the viral load of the PCV2 genome by using an ISH technique. Animals of the PCV2 positive group showed a significant downshift of the CD8(+) and DP cell subsets compared to the other groups (P < 0.05). Moreover, in PCV2-positive pigs, the amount of PCV2 genome in lymphoid tissues was related to the degree of cell depletion in those tissues (P < 0.05) as well as to the relative decrease in IgM(+) and CD8(+) cells in peripheral blood. These data support the notion that PCV2-positive pigs might have an impaired immune response. PMID- 11874859 TI - Differential modulation of surface and intracellular protein expression by T cells after stimulation in the presence of monensin or brefeldin A. AB - Intracellular cytokine staining is an increasingly popular analytical tool that can be used to define the profile of cytokines in various disease states. One important requirement for this assay is the inclusion of a protein transport inhibitor in stimulated cell cultures to trap the cytokine, thus allowing a brighter signal. Two compounds commonly used for this purpose are brefeldin A (BFA) and monensin (MN). Flow cytometry was used to assess the differential effects of BFA and MN on surface CD3, -4, -8, and -69 expression and the intracellular expression of gamma interferon (IFN-gamma) and tumor necrosis factor alpha (TNF-alpha) following stimulation with phorbol myristate acetate and ionomycin. We found that BFA blocked the majority of CD3(+) cells from expressing surface CD69, but BFA did not inhibit intracellular CD69 expression. MN did not significantly inhibit surface CD69 expression. With regard to lymphocyte marker expression following activation, surface CD4 expression was significantly downregulated; however, less downregulation was observed with BFA treatment than with MN treatment. Analyzing intracellular cytokine expression, BFA trapped a greater percentage of TNF-alpha inside activated cells than MN. An analysis of the cytokine concentration in culture supernatants indicated that cells treated with MN released TNF-alpha and IFN-gamma from the cells, while the BFA-treated cells released IFN-gamma only. With prolonged (18-h) stimulation, the cells treated with MN were less viable than those treated with BFA. We conclude that the choice of a protein transport inhibitor is an important variable in this assay. When developing this method as a tool for clinical immunology laboratory analysis, investigators should consider the differential effects of BFA and MN on results. PMID- 11874860 TI - T-cell-mediated immune responses in patients with cutaneous or mucosal leishmaniasis: long-term evaluation after therapy. AB - T-cell immune responses in patients with cutaneous leishmaniasis (CL) and mucosal leishmaniasis (ML) were studied during the active disease, at the end of therapy, and 1 to 17 years posttherapy (long-term follow-up). Lymphocyte proliferative responses, phenotypic characterization of CD4(+) and CD8(+) Leishmania-reactive T cells, and cytokine production were assayed. Patients with active ML and CL showed higher proportions of CD4(+) than CD8(+) T cells. In CL, the healing process was associated with a decrease of CD4(+) and an increase of CD8(+), leading to similar CD4(+) and CD8(+) proportions. This pattern was only seen in ML after long-term therapy. Long-term follow-up of patients with CL showed a positive CD4(+)/CD8(+) ratio as observed during the active disease, although the percentages of these T cell subsets were significantly lower. Patients with CL did not show significant differences between gamma interferon (IFN-gamma) and interleukin-5 (IL-5) production during the period of study. Patients with active ML presented higher IFN-gamma and IL-5 levels compared to patients with active CL. IL-4 was only detected during active disease. Patients long term after cure from ML showed increasing production of IFN-gamma, significant decrease of IL-5, and no IL-4 production. Two apparently beneficial immunological parameters were detected in tegumentary leishmaniasis: (i) decreasing proportions of CD4(+) Leishmania-reactive T cells in the absence of IL-4 production associated with cure of CL and ML and (ii) decreasing levels of IL-5 long after cure, better detected in patients with ML. The observed T-cell responses maintained for a long period in healed patients could be relevant for immunoprotection against reinfection and used as a parameter for determining the prognosis of patients and selecting future vaccine preparations. PMID- 11874861 TI - Improved assessment of T-cell receptor (TCR) VB repertoire in clinical specimens: combination of TCR-CDR3 spectratyping with flow cytometry-based TCR VB frequency analysis. AB - Antigen-specific T-cell responses may be described by combining three categories: (i) the specificity and effector functions of a T-cell population, (ii) the quantity of T-cell responses (i.e., the number of responding T cells within the CD4/CD8 population), and (iii) the "quality" of T cells (defined by the T-cell receptor [TCR] structure). Several methods to measure T-cell responses are now available including evaluation of T-cell precursors using limiting dilution, the enzyme-linked immunospot assay, ex vivo TCR variable (v)-segment analysis determined by flow cytometry, and TCR-CDR3 length analysis (spectratyping), as well as identification of peptide-specific T cells using major histocompatibility complex (MHC) class I tetramers containing appropriate peptides. Until now, only a limited set of MHC-peptide complexes have been available as tetramer complexes. We demonstrate that CD8(+) or CD4(+) T cells in patients with cancer can be molecularly defined using a combination of spectratyping (TCR structure and "molecular composition") plus the implementation of an antibody panel directed against 21 individual VB TCR chains ("quantity" of T-cell families). This approach is instrumental in defining and comparing the magnitudes of CD4(+) or CD8(+) T-cell responses over time in individual patients, in comparing the TCR VA and VB repertoire in different anatomic compartments, and in comparing the TCR VA VB diversity with that in normal healthy controls. This method provides the means of objectively defining and comparing the TCR repertoire in patients undergoing vaccination protocols and underlines the necessity to calibrate the TCR-CDR3 analysis with a qualitative assessment of individual TCR VB families. PMID- 11874863 TI - Immunophenotyping of peripheral blood lymphocytes in Saudi men. AB - Flow cytometry is an important tool for the diagnosis and follow-up of immunodeficiency patients, as well as for patients with leukemia and lymphoma. Lymphocytes and their subsets show variations with race. The aim of this study was to establish reference ranges for lymphocytes and their subsets in an Saudi adult population by using flow cytometry. Blood samples obtained from 209 healthy Saudi men were used for this study. All blood donors were between 18 and 44 years old. Lymphocytes and their subsets were analyzed by flow cytometry, and the absolute and percentage values were calculated. We investigated the expression of T-cell markers (CD3, CD4, and CD8), B cells (CD19), and natural killer cells (CD16 and CD56). The absolute and percent values of each cell subset were compared with published data from different populations by using the Student t test. Reference ranges, each expressed as the mean +/- the standard deviation, were as follows: leukocytes (6,335 +/- 1759), total lymphocytes (2,224 +/- 717), CD3 cells (1,618 +/- 547), CD4 cells (869 +/- 310), CD8 cells (615 +/- 278), CD19 cells (230 +/- 130), and CD3-CD16(+)/CD56+ cells (262 +/- 178). The CD4/CD8 ratio was 1.6 +/- 0.7. Our results for B cells, CD4 cells, and CD8 cells and for the CD4/CD8 ratio fell in between the reported results for Ethiopian and Dutch subjects. Our results were also different from previously reported findings in an Saudi adult population that showed no increase in CD8 T cells. We thus establish here the reference ranges for lymphocytes and their subsets in a large cohort of Saudi men. The CD8 cell count was not abnormally high, as previously reported, and fell in between previous results obtained for African and European populations. PMID- 11874862 TI - Antigen-driven T-cell selection in patients with cervical cancer as evidenced by T-cell receptor analysis and recognition of autologous tumor. AB - We characterized the T-cell receptor (TCR) repertoire in freshly harvested tumor lesions, in short-term-expanded CD4(+) tumor infiltrating lymphocytes (TIL) as well as in CD4(+) and CD8(+) peripheral blood lymphocytes (PBL) from three patients with cervical cancer. Skewing of the T-cell repertoire as defined by measuring the length of the complementarity-determining region 3 (CDR3) of the TCR VA and VB chains was observed in CD8(+) PBL, in freshly harvested tumor tissue, as well as in CD4(+) TIL. Comparative analysis of the TCR repertoire revealed unique monoclonal TCR transcripts within the tumor lesion which were not present in PBL, suggesting selection of TCR clonotypes due to antigenic stimulation. TCR repertoire analysis of the short-term (7-day) CD4(+) TIL lines revealed that the TCR composition is markedly different from that in CD4(+) PBL or in the freshly harvested tumor tissue. Only one-third of CD4(+) TIL lines showed HLA-DR-restricted recognition of autologous tumor cells as defined by cytolysis. These data provide support for the antigen-driven selection of T cells within cervical cancer lesions and suggest that analysis of the TCR repertoire may aid in obtaining an objective description of the immune response in patients with cervical cancer who are undergoing epitope-based immunotherapy. PMID- 11874864 TI - Impaired bactericidal activity and host resistance to Listeria monocytogenes and Borrelia burgdorferi in rats administered an acute oral regimen of ethanol. AB - A rat model was used to examine how ethanol ingestion may interfere with antimicrobial immunity both in vitro and in vivo. Nonimmune Long-Evans rats were given a short-course treatment orally with excessive amounts of ethanol. Their spleens were removed at the time of sacrifice, and separate spleen cell suspensions were prepared and tested in vitro for their ability to kill two bacterial pathogens, Listeria monocytogenes and Borrelia burgdorferi. After the bacteria were mixed separately with various concentrations of spleen cells, it was found that spleen cells from the ethanol-treated rats killed fewer bacteria than matching pair-fed controls, based on counts of the number of cultured CFU (for Listeria) or based on microscopic examination (for Borrelia). For the in vivo studies, ethanol-treated and control rats were infected intraperitoneally with Listeria, and then, 1 to 3 days later, they were assessed for systemic infection based on the numbers of organisms present in their livers and spleens. Numbers of bacterial CFU for both organs were significantly higher in the group fed ethanol for the first 2 days after listerial challenge. These results support the concept that acute exposure to high levels of ethanol can impair host defense mechanisms, especially those expressed at the cellular level, which could lead to increased susceptibility to certain types of infections. PMID- 11874865 TI - Exposure of HEp-2 cells to stress conditions influences antinuclear antibody reactivity. AB - This study of stress-related antinuclear antibody (ANA) reactivity was undertaken with the objective of improving clinical ANA testing. ANA was determined by parallel enzyme-linked immunosorbent assays of crude nuclear protein antigen extracted from HEp-2 cells either grown under optimal conditions (providing nonstress ANA antigen) or exposed to stress (providing stress ANA antigen). The stress stimuli used were gamma radiation (causing DNA damage) and a hypertonic environment (causing apoptosis). Signs of stress-related ANA reactivity were seen among connective tissue disease (CTD) patients (including patients with systemic lupus erythematosus; mixed CTD; calcinosis, Reynaud's phenomenon, esophageal motility disorders, sclerodactyly, and telangiectasia; scleroderma; and Sjogren's syndrome): 11% showed stress-positive ANA (i.e., a significantly stronger ANA reactivity with the extract from stressed cells), whereas 21% showed a markedly weaker reaction with the stress antigen. In contrast, among ANA screening patient sera, with no diagnosis of CTD, the fraction showing stress-positive ANA was higher (7 to 8%, depending on the type of stress) than among those showing a lower reactivity with stress antigen (1.5 to 2.5%). Only one serum among 89 (1%) tested sera from healthy individuals showed a stress-related ANA reaction. This demonstration of stress-related ANA suggests a means to improve the performance of clinical ANA testing. PMID- 11874866 TI - Influence of specimen age and use of different negative controls in determination of intracytoplasmic levels of cytokines after whole-blood culture assay. AB - Intracytoplasmic detection of cytokines by flow cytometry has become a powerful tool in the characterization of cytokine-producing cells. However, it is not known to what extent specimen age and the use of various negative controls may influence the amount of cytokine-positive cells. We therefore compared different times of storage and the use of several negative controls in the determination of intracytoplasmic levels of cytokines. There was a substantial decline of interleukin-2- and gamma interferon-positive lymphocytes after 20 h and especially after 48 h of storage. The precision of intracytoplasmic interleukin-6 determination decreases after long-term storage compared to 2 h of storage, whereas the amount of interleukin-8-positive monocytes remained rather stable. Therefore, we recommend performing the analysis as fast as possible after the blood sample is drawn. Under consideration of isotype-matched antibodies and nonstimulated cells as negative controls instead of the purified antibody blocking control, strikingly higher amounts of interleukin-2-, gamma interferon, interleukin-6-, and interleukin-8-positive cells were found. For a meaningful interpretation of data these differences have to be kept in mind. Further studies should evaluate the exact specificity of these controls. PMID- 11874867 TI - Comparative and prospective study of different immune parameters in healthy subjects at risk for tuberculosis and in tuberculosis patients. AB - It has not been fully elucidated which of the components of the immune response against Mycobacterium tuberculosis is indicative of resistance or susceptibility. The aim of this study was to identify an immune parameter that could be indicative of either resistance or susceptibility to M. tuberculosis infection. We prospectively studied (three determinations, at months 0, 8, and 12) 15 patients with chronic pulmonary tuberculosis and 42 healthy individuals with a recent and frequent contact with tuberculosis patients. Peripheral blood mononuclear cells were stimulated with a whole-protein extract or the 30-kDa antigen of M. tuberculosis for 6 days, and several immune parameters were determined. No consistent differences between tuberculosis patients and healthy controls were detected in most immune parameters studied, including the expression of different activation antigens, cytokine secretion, lymphocyte proliferation, and nitric oxide production. However, the synthesis of tumor necrosis factor alpha, the intracellular detection of gamma interferon, and the apoptosis of monocytes under certain culture conditions tended to show clear-cut differences in cells from patients and controls (P < 0.05 in all cases for most determinations). Nevertheless, when results were analyzed on an individual basis, it was evident that a significant degree of overlapping of values from patients and controls occurred for all parameters studied. We conclude that although the immune parameters tested do not allow the identification of individuals susceptible to M. tuberculosis, the specificity and sensitivity of some of them could be improved through future studies. PMID- 11874868 TI - Immunoglobulin G, A, and M responses in serum and circulating immune complexes elicited by the 16-kilodalton antigen of Mycobacterium tuberculosis. AB - The 16-kDa cytosolic antigen of M. tuberculosis was purified to homogeneity by molecular sieving chromatography, and the diagnostic potential of the antigen was evaluated in various categories of patients by enzyme-linked immunosorbent assay (ELISA). The immunoglobulin G (IgG), IgA, and IgM antibody levels to 16-kDa antigen were estimated in the two polar groups, namely, smear- and culture positive pulmonary tuberculosis (S(+)C(+)) patients and healthy subjects (HS). Sensitivities of 62, 52 and 11% with specificities of 100, 97, and 95% were obtained for the three isotypes, respectively. The total number of positives by a combination of the three isotypes was analyzed in the polar groups, and the sensitivity improved to 83% with a specificity of 93%. Even when a combination of IgG and IgA alone was considered, the sensitivity was 82% with a specificity of 97%. Polyethylene glycol precipitation of the circulating immune complex (CIC) in sera was carried out. The CIC-bound antibodies to 16-kDa antigen were assessed by ELISA in the S(+)C(+), S(-)C(+), and S(-)C(-) categories of patients. Measuring the IgG-IgA-IgM combination positivities of the CIC-bound antibodies gave sensitivities of 97.5, 100, and 45.3%, respectively. The specificity of the assay with these combinations was maintained at 95.4%. PMID- 11874870 TI - Cell-associated interleukin-8 in cord blood of term and preterm infants. AB - To assess the effect of gestational age and labor on the interleukin-8 (IL-8) concentration in whole cord blood and serum, IL-8 levels were determined simultaneously in cord blood serum and lysate in 134 infants. Following the elimination of some of the samples due to exclusion criteria, the data for 99 uninfected infants (71 term and 28 preterm) and 9 infants with neonatal bacterial infection delivered either vaginally or by elective or emergency cesarean section were analyzed. The effects of labor and gestational age were tested by analysis of variance. IL-8 was not detectable in the serum of 25 infants, whereas IL-8 levels in whole blood were measurable in all of the samples. The median IL-8 conncentrations in whole cord blood lysate were 106 pg/ml (range, 20 to 415 pg/ml) in preterm infants and 176 pg/ml (range, 34 to 1,667 pg/ml) in term infants. In contrast to the IL-8 levels in serum, IL-8 levels in whole blood were reduced after ECS. Gestational age had no independent effect on the IL-8 concentrations in either serum or whole blood; these concentrations increased in infected infants after labor. We conclude that the neonatal proinflammatory response to labor stress was more evident in the concentrations of IL-8 in whole blood than in serum. The levels of IL-8 in whole-blood lysate reflect proinflammatory stimulation in neonates and may be a useful diagnostic tool for the early diagnosis of neonatal infection. PMID- 11874869 TI - Analysis of Chlamydia pneumoniae growth in cells by reverse transcription-PCR targeted to bacterial gene transcripts. AB - Chlamydia pneumoniae is an obligate intracellular bacterium and has a unique development cycle consisting of an elementary body (EB) and reticular body (RB). EBs survive in extracellular environments as well as infect susceptible host cells. However, EBs display no measurable metabolic activity. In contrast, RBs are metabolically active and can replicate in a host cell but are noninfectious. Therefore, analysis of C. pneumoniae growth in infected cells by conventional bacterial culture may not permit sufficient information about growth of the bacteria in cells. In this study, therefore, we examined the usefulness of the reverse transcription (RT)-PCR method for analysis of bacterial transcripts to evaluate C. pneumoniae growth in HEp-2 cells because the levels of bacterial gene transcripts are known to show the metabolic activity of bacteria. The transcripts for the C. pneumoniae hsp60 gene and 16S rRNA in the cells were easily detected just after infection, followed by a marked increase. In contrast, pyk and omcB transcripts slowly increased after a latent period. The hydrocortisone treatment of C. pneumoniae-infected cells induced an increase of all bacterial transcripts tested compared with the control group. The treatment of the infected cells with the antibiotic minocycline showed a selective inhibition of bacterial gene transcripts, even though the complete inhibition of EB production determined by the bacterial culture assay was evident. These results indicate that the determination of bacterial gene transcripts by RT-PCR might be a powerful method to analyze in detail growth of C. pneumoniae in host cells, particularly altered bacterial growth caused by agents such as antimicrobials. PMID- 11874871 TI - Kinetics of antibody responses in Rickettsia africae and Rickettsia conorii infections. AB - African tick-bite fever, caused by Rickettsia africae, is the most common tick borne rickettsiosis in sub-Saharan Africa. Mediterranean spotted fever due to Rickettsia conorii also occurs in the region but is more prevalent in Mediterranean countries. Using microimmunofluorescence, we compared the development of immunoglobulin G (IgG) and IgM titers in 48 patients with African tick-bite fever and 48 patients with Mediterranean spotted fever. Doxycycline treatment within 7 days from the onset of disease significantly prevented the development of antibodies to R. africae. In patients with African tick-bite fever, the median times to seroconversion with IgG and IgM were 28 and 25 days, respectively, after the onset of symptoms. These were significantly longer by a median of 6 days for IgG and 9 days for IgM than the times for seroconversion in patients with Mediterranean spotted fever (P < 10(-2)). We recommend that sera collected 4 weeks after the onset of signs of patients with suspected African tick-bite fever should be used for the definitive serological diagnosis of R. africae infections. PMID- 11874872 TI - Performance characteristics of the PolyTiter Immunofluorescent Titration system for determination of antinuclear antibody endpoint dilution. AB - Conventional screening for circulating antinuclear antibodies (ANA) is generally performed by immunofluorescent (IF) microscopy with a 1:40 dilution of serum. Intensity of IF staining is then semiquantitated by using twofold serial dilutions, where the highest dilution in which staining intensity equals the endpoint control is expressed as an endpoint titer. The PolyTiter Immunofluorescent Titration system (Polymedco, Inc.) facilitates ANA-IF assay (IFA) testing by relating the intensity of IF staining to reference calibrators (defined in PolyTiter units), providing an endpoint titer directly from a 1:40 dilution. This study was conducted to assess the performance characteristics of the PolyTiter system. Two technologists each evaluated 10 replicates of three specimens and two controls on five sequential days. Endpoint dilution agreement (defined as +/-2 dilutions) with the reference was 100% for all controls and for all specimens by one technologist. The second reader reported agreement of 98, 88, and 100% for the low, medium, and high specimens, respectively. Analysis of PolyTiter unit values yielded between-reader, between-run, and within-run precision coefficients of variation of less than 10%. The variance component in the lot-to-lot analysis was zero, indicating all of the variation was due to run to-run differences. Overall endpoint dilution agreement between PolyTiter and serial dilution in the evaluation of 125 specimens at three sites was 90, 93, and 86%. Pattern identification with the PolyTiter was similar to that with serial dilution. The PolyTiter system demonstrates acceptable performance for routine ANA-IFA testing in the clinical laboratory. PMID- 11874873 TI - Treatment with neurokinin-1 receptor antagonist reduces severity of inflammatory bowel disease induced by Cryptosporidium parvum. AB - Inflammatory bowel disease (IBD) is a chronic, debilitating disorder of uncertain and perhaps multiple etiologies. It is believed to be due in part to disregulation of the immune system. Neuroimmune interactions may be involved in induction or maintenance of IBD. In the present study, we examined the potential role of a neurotransmitter, substance P, in a mouse model of IBD. We found that binding sites for substance P, and more specifically, neurokinin-1 receptors, were upregulated in intestinal tissue of mice with IBD-like syndrome. Dosing of mice with LY303870, a neurokinin-1 receptor antagonist, reduced the severity of IBD, and treatment of mice with preexisting IBD allowed partial healing of lesions. We hypothesize that blocking the binding of substance P to the neurokinin-1 receptor interrupts the inflammatory cascade that triggers and maintains intestinal lesions of IBD. PMID- 11874874 TI - Isolation and characterization of two European strains of Ehrlichia phagocytophila of equine origin. AB - We report the isolation and partial genetic characterization of two equine strains of granulocytic Ehrlichia of the genogroup Ehrlichia phagocytophila. Frozen whole-blood samples from two Swedish horses with laboratory-verified granulocytic ehrlichiosis were inoculated into HL-60 cell cultures. Granulocytic Ehrlichia was isolated and propagated from both horses. DNA extracts from the respective strains were amplified by PCR using primers directed towards the 16S rRNA gene, the groESL heat shock operon gene, and the ank gene. The amplified gene fragments were sequenced and compared to known sequences in the GenBank database. With respect to the 16S rRNA gene, the groESL gene, and the ank gene, the DNA sequences of the two equine Ehrlichia isolates were identical to sequences found in isolates from clinical cases of granulocytic ehrlichiosis in humans and domestic animals in Sweden. However, compared to amplified DNA from an American Ehrlichia strain of the E. phagocytophila genogroup, differences were found in the groESL gene and ank gene sequences. PMID- 11874875 TI - Immunocytochemical method for early laboratory diagnosis of tuberculous meningitis. AB - A simple immunocytochemical method was standardized for the direct demonstration of mycobacterial antigen in cerebrospinal fluid (CSF) specimens of patients with tuberculous meningitis (TBM). CSF-cytospin smears were prepared from 22 patients with a clinical diagnosis of TBM and also from an equal number of patients with nontuberculous neurological diseases (disease control). Immunocytological demonstration of mycobacterial antigens in the cytoplasm of monocytoid cells was attempted, by using rabbit immunoglobulin G to Mycobacterium tuberculosis as the primary antibody. Of the 22 CSF-cytospin smears from TBM patients, 16 showed positive immunostaining, while all of the CSF-cytospin smears from the disease control showed negative immunostaining for mycobacterial antigen. The technical aspects of this immunocytological method for the demonstration of mycobacterial antigens are simple, rapid, and reproducible, as well as specific, and therefore can be applied for the early diagnosis of TBM, particularly in patients in whom bacteriological methods did not demonstrate the presence of M. tuberculosis in the CSF. PMID- 11874876 TI - Role of Th1 and Th2 cytokines in immune response to uncomplicated Plasmodium falciparum malaria. AB - The relative balance between Th1 and Th2 cytokines appears crucial, since the role of cytokines has been evaluated in several studies by comparison of clinically heterogeneous groups of patients. The aim of this study is to determine the role of proinflammatory Th1 cytokines, interleukin-12 (IL-12) and gamma interferon (IFN-gamma), and anti-inflammatory Th2 cytokines, IL-4 and IL 10, in a homogeneous group of patients with uncomplicated Plasmodium falciparum malaria. Levels of IL-12, IFN-gamma, Il-4, and IL-10 in serum for 20 adult patients and 15 healthy control subjects were determined by an immunoenzymatic assay. Serum levels of Th1 cytokines, IL-12 (8.6 +/- 2.8 pg/ml; controls, 3.2 +/- 0.7 pg/ml) and IFN-gamma (39.2 +/- 67.6 pg/ml; controls, 8.4 +/- 6.3 pg/ml), were significantly increased at admission; 3 days later, levels of IL-12 in serum remained significantly high (8.8 +/- 2.6 pg/ml), whereas IFN-gamma levels returned to control values. The anti-inflammatory response of Th2 cytokines (IL 10 and IL-4) was distinct. Levels of IL-10 in serum were not significantly increased at day 0 and day 3 (306.6 +/- 200.4 pg/ml and 56.6 +/- 38.4 pg/ml, respectively; controls, 17.4 +/- 9.0 pg/ml). In contrast, levels of IL-4 in serum were not increased on admission (3.4 +/- 1.2 pg/ml; controls, 2.4 +/- 0.8 pg/ml), but at day 3 a moderate and significant increase of IL-4 levels was observed (4.5 +/- 1.7 pg/ml). In conclusion, the increase of Th1 cytokine IL-12 and IFN-gamma levels during the acute phase of uncomplicated P. falciparum malaria may reflect an early and effective immune response regulated by proinflammatory Th1 cytokines, and in particular IFN-gamma may play a role in limiting progression from uncomplicated malaria to severe and life-threatening complications. PMID- 11874877 TI - Relevance of dietary lipids as modulators of immune functions in cells infected with Listeria monocytogenes. AB - Nutritional status may have significant importance for the immune system, and particularly, unsaturated fatty acids may serve as modulators of immune functions. Clinical and epidemiological studies have demonstrated that fatty acids are involved in the reduction of the inflammatory processes that occur in diseases characterized by an overactivation of the immune system. At the same time, an increase in susceptibility to infection has also been reported. The importance of immune system modulation by dietary lipids in the presence of an intracellular bacterial pathogen, such as Listeria monocytogenes, was evaluated in the present study. BALB/c mice were divided into four groups which were each fed a low-fat (2.5% by weight) diet, an olive oil (OO; 20% by weight) diet, a fish oil (FO; 20% by weight) diet, or a hydrogenated coconut oil (HCO; 20% by weight) diet for 4 weeks. In each group, lymphocye proliferation was measured, and a reduction in the stimulation index was observed in the FO and HCO groups. Cytotoxicity exerted by L. monocytogenes was increased in the groups fed diets containing OO and FO after 6 h of incubation with the bacterium. An important increase in the production of reactive oxygen species was found in the groups fed the HCO diet after 12 h of incubation with L. monocytogenes. Finally, invasion and adhesion factors were not modified substantially by the action of dietary lipids, although these factors were reduced in cells from mice fed an FO diet. These results underline the importance of several dietary lipids as biological modulators of immune functions and their crucial role in the alteration of host natural resistance. PMID- 11874878 TI - Invasive pneumococcal infections in Denmark from 1995 to 1999: epidemiology, serotypes, and resistance. AB - Danish nationwide surveillance data on invasive pneumococcal disease from the 5 year period from 1995 to 1999, including 5,452 isolates, are presented and described. Annual overall incidence rates, serotype distribution, and antimicrobial susceptibility patterns of the isolates were monitored. Major changes in the total annual incidence rate from 27/100,000 in 1996 to 17/100,000 in 1999 and a significant change in the proportion of invasive isolates belonging to types 1 and 12F were observed. The serotype coverage rate by the 23-valent polysaccharide vaccine among the elderly was 92.9%, and the serotype coverage rate by the 7-, 9-, and 11-valent pneumococcal conjugate vaccines among children less than 2 years old were 71.7, 75.2, and 81.4%, respectively. Invasive isolates with reduced susceptibility to penicillin or erythromycin increased from 1995 to 1999, with a high proportion of the penicillin-nonsusceptible invasive isolates originating from people 60 years old or older (57.0%). These observations underline the importance of adequate surveillance systems of invasive pneumococcal disease to introduce and maintain national vaccine strategies and adequate antibiotic policy. PMID- 11874879 TI - Diagnostic usefulness of antibodies against ribosome recycling factor from Brucella melitensis in human or canine brucellosis. AB - The diagnostic usefulness of an enzyme-linked immunosorbent assay (ELISA) using a purified recombinant ribosome recycling factor from Brucella melitensis (CP24 antigen) was tested in human and canine infections caused by smooth and rough Brucella species, respectively. Anti-CP24 antibodies were detected in 9 (43%) of 21 consecutive cases of canine brucellosis and in 8 (53%) of 15 dogs followed for 60 days after the diagnosis of acute brucellosis. Among eight patients with acute brucellosis, anti-CP24 antibodies were detected in four in the 10 weeks following diagnosis, but the remaining four were negative during the whole follow-up (22 weeks). The frequency of anti-CP24 antibodies was also low among 24 patients with subacute brucellosis and 23 patients with chronic illness (29 and 26%, respectively). While all patients positive for anti-CP24 antibodies were also positive for antibodies to total cytoplasmic proteins of Brucella (CP), five were negative for antibodies to another cytoplasmic protein, the Brucella lumazine synthase (BLS). When a larger sample of 35 human sera negative for anti-BLS antibodies was assayed, 85.7% were positive for anti-CP24 antibodies, suggesting that the combined measurement of both reactivities could yield a higher sensitivity than any test alone. To test this hypothesis, an ELISA combining both antigens was designed. The percentage of positive results among chronic cases was higher for this assay than for the individual measurement of anti-CP24 or anti BLS antibodies (83 versus 26 and 65%, respectively) and was closer to the value obtained for anti-CP antibodies (91%). The frequency of anti-CP24 antibodies is low in both canine and human brucellosis. In the latter case, however, an ELISA combining CP24 and BLS is more sensitive than assays measuring anti-CP24 or anti BLS antibodies separately and almost as sensitive as the ELISA using CP. PMID- 11874880 TI - Evaluation of enzyme-linked immunosorbent assay for diagnosis of post-kala-azar dermal leishmaniasis with crude or recombinant k39 antigen. AB - The diagnosis of post-kala-azar dermal leishmaniasis (PKDL), a dermatosis that provides the only known reservoir for the parasite Leishmania donovani in India, remains a problem. Timely recognition and treatment of PKDL would contribute significantly to the control of kala-azar. We evaluated here the potential of the enzyme-linked immunosorbent assay (ELISA) as a diagnostic tool for PKDL. Antigen prepared from promastigotes and axenic amastigotes with parasite isolates that were derived from skin lesions of a PKDL patient gave sensitivities of 86.36 and 92%, respectively, in the 88 PKDL cases examined. The specificity of the ELISA test was examined by testing groups of patients with other skin disorders (leprosy and vitiligo) or coendemic infections (malaria and tuberculosis), as well as healthy controls from areas where this disease is endemic or is not endemic. A false-positive reaction was obtained in 14 of 144 (9.8%) of the controls with the promastigote antigen and in 14 of 145 (9.7%) of the controls with the amastigote antigen. Evaluation of the serodiagnostic potential of recombinant k39 by ELISA revealed a higher sensitivity (94.5%) and specificity (93.7%) compared to the other two antigens used. The data demonstrate that ELISA with crude or recombinant antigen k39 provides a relatively simple and less invasive test for the reliable diagnosis of PKDL. PMID- 11874881 TI - Heterologous expression, purification, and immunological reactivity of a recombinant HSP60 from Paracoccidioides brasiliensis. AB - The complete coding cDNA of HSP60 from Paracoccidioides brasiliensis was overexpressed in an Escherichia coli host to produce high levels of recombinant protein. The protein was purified by affinity chromatography. A total of 169 human serum samples were tested for reactivity by Western blot analysis with the purified HSP60 recombinant protein. Immunoblots indicated that the recombinant P. brasiliensis HSP60 was recognized by antibodies in 72 of 75 sera from paracoccidioidomycosis patients. No cross-reactivity was detected with individual sera from patients with aspergillosis, sporotrichosis, cryptococcosis, and tuberculosis. Reactivity to HSP60 was observed in sera from 9.52% of control healthy individuals and 11.5% of patients with histoplasmosis. The high sensitivity and specificity (97.3 and 92.5%, respectively) for HSP60 suggested that the recombinant protein can be used singly or in association with other recombinant antigens to detect antibody responses in P. brasiliensis-infected patients. PMID- 11874882 TI - Development and evaluation of an in-house enzyme-linked immunosorbent assay for early diagnosis and monitoring of human pythiosis. AB - Human pythiosis is an emerging, fatal, infectious disease caused by Pythium insidiosum and occurs in both tropical and subtropical countries. Thalassemic patients, farmers, and aquatic-habitat residents are predisposed to this disease. Delayed treatment due to the long time required for isolation and identification of the causative organism, as well as the difficulty in obtaining internal organ specimens, results in high morbidity and mortality. To facilitate rapid diagnosis, an in-house enzyme-linked immunosorbent assay (ELISA) for the detection of immunoglobulin G antibodies against P. insidiosum was developed and evaluated for the diagnosis and monitoring of human pythiosis. Sixteen sera were collected from seven culture-proven human pythiosis cases. A total of 142 sera from thalassemic patients, from patients with other infectious diseases, and from healthy blood donors served as controls. All sera were tested in duplicate. By choosing a suitable cutoff point to maximize sensitivity and specificity, sera from pythiosis cases were all determined to be positive, whereas sera from control groups were all determined to be negative. ELISA signals from serial samples of sera taken from treated patients showed gradually declining levels of antibodies to P. insidiosum. The ELISA test was highly sensitive (100%) and specific (100%) and was useful for early diagnosis and for monitoring the treatment for pythiosis. PMID- 11874883 TI - Sensitive microplate assay for detection of bactericidal antibodies to Vibrio cholerae O139. AB - A microplate assay for the detection of bactericidal antibodies to Vibrio cholerae O139 is described. The assay is sensitive, highly reproducible, specific, and convenient to perform. It has been used to demonstrate the induction of serum bactericidal antibodies in Vietnamese recipients of an oral, inactivated, bivalent O1/O139 vaccine, as well as in Bangladeshi patients with O139 disease. In both study groups there was a significant inverse correlation between the preexposure level of antibodies in serum and the magnitude of the subsequent bactericidal response. Although infection generated stronger responses than vaccination, the proportion of responders was similar among individuals with low background titers. PMID- 11874884 TI - Neutralizing antibody responses to human herpesviruses 6 and 7 do not cross-react with each other, and maternal neutralizing antibodies contribute to sequential infection with these viruses in childhood. AB - Seroprevalence of human herpesvirus 6 (HHV-6) and HHV-7 infections is very high throughout the world, and almost all people are exposed first to HHV-6 and second to HHV-7 in their childhood. However, it is not clear whether the neutralizing (NT) antibody response between each virus is cross-reactive or not. To elucidate the NT antibody response between each virus, 55 serum samples from an adult group (subjects 22 to 88 years old) and 60 serum samples from a young group (subjects 2 to 18 years old) were examined by a dot blot method for detecting viral late antigen. Thirty-nine serum samples obtained from cord bloods and a few serum samples obtained from pediatric patients with exanthem subitum were also examined to assess the maternal transferred NT antibodies against each virus. The NT antibody titers against HHV-7 in the adult group remained high throughout all the individuals, and none were negative. Those against HHV-6 were high values in the young group but low values, including negative values (three samples), in the adult group. These results suggested that the NT antibody response to either HHV 6 or HHV-7 in each individual was specific to each virus and did not cross-react with each other. In the adult group, the NT antibody response to HHV-6 decreased, while that to HHV-7 remained high throughout all the individuals. Maternal transferred NT antibody titers against HHV-7 were higher and remained longer after birth than those of HHV-6, and these findings were in accord with the clinical observation that HHV-6 infection usually occurs earlier than HHV-7 infection. PMID- 11874885 TI - Elucidation of the cross-reactive immunoglobulin M response to human herpesviruses 6 and 7 on the basis of neutralizing antibodies. AB - Human herpesvirus 6 (HHV-6) is closely related to HHV-7 in terms of genome organization and sequence. The cross-reactive responses between HHV-6 and HHV-7 have been reported by using immunofluorescent techniques. Recently we have shown that neutralizing (NT) antibody responses are specific to each virus and do not cross-react. We took advantage of this and used the NT antibody response to estimate the time of seroconversion to each virus and examined the pattern of humoral immune response, especially the immunoglobulin M (IgM) response, against each virus antigen in the natural course of infection with HHV-6 and HHV-7. In children who experienced HHV-6 infection first, followed by HHV-7 infection, the IgM response at the first HHV-6 infection was directed only against HHV-6, while no IgM response was directed against HHV-7 at the second HHV-7 infection. In contrast, in children who experienced HHV-7 infection first, followed by HHV-6 infection, the IgM response at the first HHV-7 infection was directed not only against HHV-7 but also against HHV-6. These data suggest that cross-reactive responses to heterologous viruses should be taken into consideration when making a diagnosis based on IgM antibody. PMID- 11874886 TI - Dependence of bacterial protein adhesins on toll-like receptors for proinflammatory cytokine induction. AB - Toll-like receptors (TLRs) are important signal transducers that mediate inflammatory reactions induced by microbes through pattern recognition of virulence molecules such as lipopolysaccharide (LPS) and lipoproteins. We investigated whether proinflammatory cytokine responses induced by certain bacterial protein adhesins may also depend on TLRs. In differentiated THP-1 mononuclear cells stimulated by LPS-free recombinant fimbrillin (rFimA) from Porphyromonas gingivalis, cytokine release was abrogated by monoclonal antibodies (MAbs) to CD14 and TLR4 but not to TLR2. Similar experiments using anti-beta2 integrin MAbs suggested that beta2 integrins (CD11/CD18) also play a role in cytokine induction by rFimA or native fimbriae. Minor fimbriae (distinct from the fimA-encoded major fimbriae) of P. gingivalis induced proinflammatory cytokine release in a CD14- and TLR2-dependent mode. Cytokine induction by BspA, a leucine rich repeat protein from Bacteroides forsythus, depended heavily on CD14 and TLR2. We also found that the ability of the streptococcal protein AgI/II to stimulate cytokine release depended partially on CD14 and TLR4, and the AgI/II segment that possibly interacts with these receptors was identified as its N terminal saliva-binding region. When THP-1 cells were exposed to rFimA for 24 h, surface expression of CD14 and CD18 was decreased and the cells became hyporesponsive to cytokine induction by a second challenge with rFimA. However, tolerance induction was abolished when the THP-1 cells were pretreated with rFimA in the presence of either anti-CD14 MAb or anti-TLR4 MAb. Induction of cross tolerance between rFimA and LPS correlated with downregulation of the pattern recognition receptors involved. Our data suggest that the CD14-TLR2/4 system is involved in cytokine production and tolerance induction upon interaction with certain proinflammatory bacterial protein adhesins. PMID- 11874887 TI - Reliability of Helicobacter pylori and CagA serological assays. AB - Background serological assays for Helicobacter pylori are commonly used without knowledge of reliability. This information is needed to define the ability of serological tests to determine either new cases of infection or loss of infection in longitudinal studies. We evaluated the reproducibility and the interrelationships of serological test results for H. pylori and cytotoxin associated gene product A (CagA) enzyme-linked immunoassays within a subset of participants in a population-based study. Stored samples from 1,229 participants in the third U.S. National Health and Nutrition Examination Survey were replicate serologically tested for H. pylori and CagA. Overall disagreement was 3.4% between duplicate tests for H. pylori (or 2.3% if equivocal results were disregarded). Six percent of samples positive on the first test had an immune serum ratio at least 30% lower on repeat testing. The odds ratio for H. pylori seropositivity on retesting was 2.8 (95% confidence interval [CI] = 1.8 to 4.5) when CagA serology was positive versus when it was negative. CagA antibody was found among 47.8% of H. pylori-equivocal and 7.0% of H. pylori-negative samples. CagA-positive yet H. pylori-negative samples were more likely to occur among Mexican Americans (odds ratio, 5.2; 95% CI = 2.4 to 11.4) and non-Hispanic blacks (odds ratio, 5.5; 95% CI = 2.3 to 13.0) than among non-Hispanic whites. Relying on repeated H. pylori serological tests over time to determine infection rates may result in misinterpretation due to limits in test reproducibility. CagA testing may have a role in verifying infection. PMID- 11874888 TI - Selective in vivo depletion of CD4(+) T lymphocytes with anti-CD4 monoclonal antibody during acute infection of calves with Anaplasma marginale. AB - To investigate the in vivo role of CD4(+) T lymphocytes during acute anaplasmosis, thymectomized calves were selectively depleted of CD4(+) T lymphocytes by treatment with anti-CD4 monoclonal antibody (MAb) and were then infected with the Florida strain of Anaplasma marginale in two sequential experiments (experiments 1 and 2). Treatment of thymectomized calves with a total of 5.0 mg of anti-CD4 MAb/kg of body weight during the 1st week followed by 0.3 mg/kg doses administered twice weekly for 7 weeks resulted in significant depletion of CD3(+) CD4(+) and CD4(+) CD45R(+) (naive) T lymphocytes from blood, spleen, and peripheral lymph nodes for the duration of the 8-week study, compared to the results for thymectomized control calves treated with a subclass-matched MAb. All calves became parasitemic and pyretic following experimental infection with A. marginale, and decreases in packed cell volume (PCV) coincided with peak parasitemia. No significant differences in PCV or parasitemia were observed between treatment groups. Thymectomized calves treated with anti-CD4 MAb were able to mount an anti-A. marginale antibody response, although in experiment 2, anti-CD4 MAb-treated calves had four- to sixfold lower immunoglobulin G1 (IgG1) and no detectable IgG2 anti-A. marginale major surface protein 2-specific antibody titers compared to thymectomized control calves treated with a subclass matched MAb. At the level of CD4(+)-T-lymphocyte depletion achieved and experimental anaplasmosis induced, thymectomized anti-CD4 MAb-treated calves were able to control acute anaplasmosis. This was in contrast to the prediction that significant depletion of CD4(+) T lymphocytes would abrogate resistance to acute infection. PMID- 11874890 TI - Granulocyte colony-stimulating factor improves deficient in vitro neutrophil transendothelial migration in patients with advanced liver disease. AB - Bacterial infections are frequent complications in patients with liver cirrhosis. Cirrhotic patients present abnormalities in both innate and adaptive immune responses, including a deficient neutrophil recruitment to infected sites. The purpose of this study was to assess neutrophil-endothelium interactions in cirrhotic patients and evaluate the effects of G-CSF on this process. We studied neutrophil adhesion and transendothelial migration in 14 cirrhotic patients and 14 healthy controls. We also analyzed neutrophil expression of the adhesion molecules CD62L and CD11b in whole blood by flow cytometry. Cirrhotic patients expressed higher levels of CD11b than healthy controls, whereas CD62L expression was significantly lower, suggesting exposure of neutrophils to activating agents within the bloodstream. Neutrophils from cirrhotic patients showed increased adhesion to both resting and tumor necrosis factor alpha-stimulated microvascular endothelial cells and decreased transendothelial migration. Granulocyte colony stimulating factor (G-CSF) (100 ng/ml) significantly enhanced neutrophil adhesion to microvascular endothelial cells in healthy controls but not in cirrhotic patients. G-CSF also significantly improved neutrophil transmigration in cirrhotic patients and healthy controls. In conclusion, cirrhotic patients exhibit increased neutrophil adhesion to microvascular endothelium and deficient transendothelial migration. G-CSF enhances neutrophil transendothelial migration in cirrhotic patients despite having no effect on neutrophil adhesion. Therefore, G-CSF may be able to increase neutrophil recruitment into infected sites in these patients. PMID- 11874891 TI - Elevated levels of lipopolysaccharide-binding protein and soluble CD14 in plasma in neonatal early-onset sepsis. AB - No data on lipopolysaccharide-binding protein (LBP) in newborns with sepsis have been available up to now. We therefore determined levels of LBP and soluble CD14 (sCD14) in plasma of healthy and septic neonates in order to evaluate their potential diagnostic role. The study included prospectively collected patient samples of two recently published studies on cytokine expression in neonatal sepsis. Twenty-nine septic patients were enrolled in the present analysis. Samples--either cord blood or peripheral blood--from patients admitted within the first 24 h of life for suspicion of sepsis and cord blood samples of a control group of 40 healthy mature infants delivered spontaneously were analyzed. For seven patients of the septic group, a second sample collected between 24 and 48 h of life was available. Levels of sCD14 and LBP in plasma were determined by an enzyme immunoassay using recombinant CD14 and LBP as standards. LBP and sCD14 were correlated to cytokine plasma levels. In septic neonates, LBP (median, 36.6 versus 7.8 microg/ml; P < 0.001) and sCD14 (median, 0.42 versus 0.28 microg/ml; P < 0.001) levels were highly elevated when compared to those of healthy neonates and strongly correlated to granulocyte colony-stimulating factor (G-CSF), interleukin-1beta (IL-1beta), IL-6, and IL-8 levels. LBP levels in septic neonates analyzed between 24 and 48 h of life even increased when compared to samples obtained at or shortly after delivery (median, 36.6 versus 60 microg/ml; P = 0.038). In summary, levels of LBP in plasma of neonates with early-onset sepsis are significantly elevated; the elevated plasma levels seem to persist for more than 24 h, which could provide the clinician with a prolonged time period to identify the newborn with bacterial sepsis. PMID- 11874889 TI - Systemic inflammation in cardiovascular and periodontal disease: comparative study. AB - Epidemiological studies have implicated periodontal disease (PD) as a risk factor for the development of cardiovascular disease (CVD). These studies addressed the premise that local infection may perturb the levels of systemic inflammatory mediators, thereby promoting mechanisms of atherosclerosis. Levels of inflammatory mediators in the sera of subjects with only PD, only CVD, both diseases, or neither condition were compared. Subjects were assessed for levels of C-reactive protein (CRP), serum amyloid A (SAA), ceruloplasmin, alpha(1)-acid glycoprotein (AAG), alpha(1)-antichymotrypsin (ACT), and the soluble cellular adhesion molecules sICAM-1 and sVCAM by enzyme-linked immunoabsorbent and/or radial immunodiffusion assays. CRP levels in subjects with either condition alone were elevated twofold above subjects with neither disease, whereas a threefold increase was noted in subjects with both diseases (P = 0.0389). Statistically significant increases in SAA and ACT were noted in subjects with both conditions compared to those with one or neither condition (P = 0.0162 and 0.0408, respectively). Ceruloplasmin levels were increased in subjects with only CVD (P = 0.0001). Increases in sVCAM levels were noted in all subjects with CVD (P = 0.0054). No differences in sICAM levels were noted among subject groups. A trend toward higher levels of AAG was noted in subjects with both conditions and for ACT in subjects with only PD. Immunohistochemical examination of endarterectomy specimens of carotid arteries from subjects with atherosclerosis documented SAA and CRP deposition in association with atheromatous lesions. The data support the hypothesis that localized persistent infection may influence systemic levels of inflammatory mediators. Changes in inflammatory mediator levels potentially impact inflammation-associated atherosclerotic processes. PMID- 11874892 TI - Newly characterized species-specific immunogenic Chlamydophila pneumoniae peptide reactive with murine monoclonal and human serum antibodies. AB - A monoclonal antibody (MAb) directed against an unknown Chlamydophila pneumoniae epitope has been characterized, and the respective peptide mimotope has been identified. A murine MAb specific for C. pneumoniae was used to select peptides from phage display libraries. The peptides identified from the phage display library clones reacted specifically with the respective target murine MAb and with human sera previously identified as having antibody titers to C. pneumoniae. The selected peptide mimotope sequences tended to be composed of charged residues surrounding a core of hydrophobic residues. The peptide with the best binding could inhibit >95% of binding to the MAb, suggesting that the selected peptide binds the paratope of the respective MAb. The peptide reacted with human sera previously determined by microimmunofluorescence to have anti-C. pneumoniae antibodies. The peptide was competitively competed with the MAb against Renografin-purified, sonicated C. pneumoniae in an enzyme-linked immunosorbent assay and with whole-cell C. pneumoniae in an indirect fluorescence assay format, demonstrating its potential utility in the development of diagnostics. The use of this novel peptide may allow investigators to establish standardized assays free from cross-reactive Chlamydia trachomatis and Chlamydophila psittaci epitopes and immunoreactivity. PMID- 11874893 TI - Interpretation of the gamma interferon test for diagnosis of subclinical paratuberculosis in cattle. AB - A group of 252 cattle without clinical signs of paratuberculosis (paraTB) in 10 herds infected with paraTB and a group of 117 cattle in 5 herds without paraTB were selected. Whole-blood samples were stimulated with bovine, avian, and johnin purified protein derivative (PPD) and examined for gamma interferon (IFN-gamma) release. For diagnosis of paraTB, satisfactory estimated specificities (95 to 99%) could be obtained by johnin PPD stimulation irrespective of interpretation relative to bovine PPD or no-antigen stimulation alone, but numbers of test positives in the infected herds varied from 64 to 112 with different interpretation criteria. For a limited number of test-positive animals, no change in the test results could be observed with increasing antigen concentrations but IFN-gamma responses were significantly reduced (P < 0.0001) and four out of seven reactors tested negative when stimulation was performed on day-old samples. Denmark is free of bovine tuberculosis, but cross-reactivity with paraTB could be documented for cattle more than 14 months old in paraTB-infected herds compared with those in non-paraTB-infected herds. In both paraTB-free and paraTB-infected herds, false positives were observed when the test was applied to calves less than 15 months of age. Until novel antigen formulations more specific for these diseases are available, interpretation of the IFN-gamma test must be individually adjusted to fit specific needs and the context within which the test is applied and, for paraTB, the test seems most appropriate for use as a supportive tool for evaluation of disease-preventive measures in young stock. PMID- 11874894 TI - Detection of Cryptococcus neoformans DNA in tissue samples by nested and real time PCR assays. AB - Two PCR protocols targeting the 18S rRNA gene of Cryptococcus neoformans were established, compared, and evaluated in murine cryptococcal meningitis. One protocol was designed as a nested PCR to be performed in conventional block thermal cyclers. The other protocol was designed as a quantitative single-round PCR adapted to LightCycler technology. One hundred brain homogenates and dilutions originating from 20 ICR mice treated with different azoles were examined. A fungal burden of 3 x 10(1) to 2.9 x 10(4) CFU per mg of brain tissue was determined by quantitative culture. Specific PCR products were amplified by the conventional and the LightCycler methods in 86 and 87 samples, respectively, with products identified by DNA sequencing and real-time fluorescence detection. An analytical sensitivity of 1 CFU of C. neoformans per mg of brain tissue and less than 10 CFU per volume used for extraction was observed for both PCR protocols, while homogenates of 70 organs from mice infected with other fungi were PCR negative. Specificity testing was performed with genomic DNA from 31 hymenomycetous fungal species and from the ustilaginomycetous yeast Malassezia furfur, which are phylogenetically related to C. neoformans. Twenty-four strains, including species of human skin flora like M. furfur and Trichosporon spp., were PCR negative. Amplification was observed with Cryptococcus amylolentus, Filobasidiella depauperata, Cryptococcus laurentii, and five species unrelated to clinical specimens. LightCycler PCR products from F. depauperata and Trichosporon faecale could be clearly discriminated by melting curve analysis. The sensitive and specific nested PCR assay as well as the rapid and quantitative LightCycler PCR assay might be useful for the diagnosis and monitoring of human cryptococcal infections. PMID- 11874895 TI - Grape seed extract activates Th1 cells in vitro. AB - Although flavonoids manifest a diverse range of biological activities, including antitumor and antiviral effects, the molecular mechanisms underlying these activities await elucidation. We hypothesize that the flavonoid constituents of a proprietary grape seed extract (GSE) that contains procyandins exert significant antiviral and antitumor effects, by inducing production of the Th1-derived cytokine gamma interferon (IFN-gamma) by peripheral blood mononuclear cells) from healthy donors. Our results show that GSE significantly induced the transcription of IFN-gamma mRNA as demonstrated by reverse transcription-PCR but had no effect on the Th2-derived cytokine interleukin-6. The enhancing effect of GSE on IFN gamma expression was further supported by a concomitant increase in the number of cells with intracytoplasmic IFN-gamma as well as the synthesis and secretion of IFN-gamma. Our results demonstrate that the potentially beneficial immunostimulatory effects of GSE may be mediated through the induction of IFN gamma. PMID- 11874896 TI - Sodium bicarbonate enhances the severity of infection in neutropenic mice orally inoculated with Listeria monocytogenes EGD. AB - Epidemiological studies have suggested an association between antacid therapy and development of listeriosis in humans. In this study we used a neutropenic mouse model to demonstrate that oral administration of sodium bicarbonate shortly before intragastric (i.g.) inoculation with Listeria monocytogenes EGD (serotype 1/2a) significantly increased the severity of the resulting systemic infection. An explanation for this observation is provided by evidence that L. monocytogenes EGD is rapidly inactivated in synthetic gastric fluid at pH below 5. A second strain of L. monocytogenes (CM [serotype 1/2b]) exhibited little ability to cause systemic infection following i.g. inoculation and was not significantly enhanced by administration of sodium bicarbonate. Strain CM was readily inactivated in synthetic gastric fluid even at pH 7. These data suggest that gastric acidity and enzymes provide some innate defense against gastrointestinal listeriosis in neutropenic mice. PMID- 11874897 TI - Potential benefit of plasma exchange in treatment of severe icteric leptospirosis complicated by acute renal failure. AB - Leptospirosis is a common zoonosis seen worldwide, but it is rare in our locality (Hong Kong). Clinical manifestations of leptospirosis are variable and may range from subclinical infection to fever, jaundice, hemorrhagic tendency, and fulminant hepato-renal failure. Severe hyperbilirubinemia and acute renal failure have been associated with high mortality. We report our experience with a patient who developed severe Weil's syndrome with marked conjugated hyperbilirubinemia and oliguric acute renal failure. These complications persisted despite treatment with penicillin and hemodiafiltration. Plasma exchange was instituted in view of the severe hyperbilirubinemia (970 micromol/liter). This was followed by prompt clinical improvement, with recovery of liver and renal function. The beneficial effects of plasma exchange could be attributed to amelioration of the toxic effects of hyperbilirubinemia on hepatocyte and renal tubular cell function. We conclude that plasma exchange should be considered as an adjunctive therapy for patients with severe icteric leptospirosis complicated by acute renal failure who have not shown rapid clinical response to conventional treatment. PMID- 11874898 TI - Opsonophagocytosis of fluorescent polystyrene beads coupled to Neisseria meningitidis serogroup A, C, Y, or W135 polysaccharide correlates with serum bactericidal activity. AB - We developed a polysaccharide-specific flow cytometric opsonophagocytic assay (OPA) for the simultaneous measurement of functional antibody to Neisseria meningitidis serogroups A, C, Y, and W135. OPA titers significantly correlated with serum bactericidal assay titers for all serogroups tested (mean r = 0.96; P < 0.001). OPA could be used in meningococcal vaccine evaluation. PMID- 11874899 TI - Preconception seroconversion and maternal seronegativity at delivery do not rule out the risk of congenital toxoplasmosis. AB - We describe two unusual cases of congenital toxoplasmosis, one occurring after preconception maternal infection with cervical adenopathies and the other occurring after maternal infection at the very end of pregnancy with maternal seronegativity at delivery. These documented cases of congenital toxoplasmosis demonstrate the value of extending the serologic monitoring period during pregnancy, according to the individual clinical context. PMID- 11874900 TI - Effect of exogenous interleukin-18 (IL-18) and IL-12 in the course of Brucella abortus 2308 infection in mice. AB - In this study we demonstrated that combined inoculation of interleukin-12 (IL-12) and IL-18 reduced the number of bacteria in the spleens of mice infected with Brucella abortus 2308 and that the effect of the treatment was mediated by an increased capability of spleen cells to produce gamma interferon at the early phase of infection. PMID- 11874901 TI - Diagnosis of Helicobacter pylori infection in children: comparison of a salivary immunoglobulin G antibody test with the [(13)C]urea breath test. AB - The prevalence of Helicobacter pylori infection in a population-based sample of 477 children (mean age plus minus standard deviation, 5.8 plus minus 0.5 years) determined by the [(13)C]urea breath test ([(13)C]UBT) was 10.7% (95% confidence interval [CI], 8.1 to 13.8%), and that determined by salivary enzyme-linked immunosorbent assay (ELISA) was 11.9% (95% CI, 9.2 to 15.2%). Compared to the [(13)C]UBT, the sensitivity and specificity of the salivary ELISA were 80.9% (95% CI, 66.3 to 90.4%) and 95.3% (95% CI, 92.7 to 97.1%), respectively. PMID- 11874902 TI - Cervical cat scratch disease lymphadenitis in a patient with immunoglobulin M antibodies to Toxoplasma gondii. AB - We report on a young patient with chronic cervical lymphadenopathy and serological and histological evidence for infection with Bartonella henselae and Toxoplasma gondii. Serological follow-up studies, including testing for avidity of Toxoplasma-specific immunoglobulin G antibodies, assisted in the determination of the cause of the acute lymphadenitis. Our results suggest that the clinical symptoms were most likely due to cat scratch disease rather than to acute toxoplasmosis. PMID- 11874903 TI - PrP(Sc) is not detected in peripheral blood leukocytes of scrapie-infected sheep: determining the limit of sensitivity by immunohistochemistry. AB - Peripheral blood leukocytes (PBLs) from scrapie-infected sheep were evaluated for the presence of PrP(Sc) by using dissociated retropharyngeal lymph node (DRLN) cells and immunohistochemistry (IHC). PrP(Sc)-positive cells were detected in 2.05% +/- 0.28% of 3 x 10(6) DRLN cells, but were not detected in 3 x 10(6) PBLs from scrapie-infected sheep. Titration of DRLN cells mixed with PBLs showed that IHC detects a minimum of 0.00205% or 60 PrP(Sc)-positive cells in 3 x 10(6) PBLs. PMID- 11874904 TI - Beta-defensin 2 in the rhesus monkey (Macaca mulatta) and the long-tailed macaque (M. fascicularis). PMID- 11874906 TI - Cues from neuroepithelium and surface ectoderm maintain neural crest-free regions within cranial mesenchyme of the developing chick. AB - Within the developing vertebrate head, neural crest cells (NCCs) migrate from the dorsal surface of the hindbrain into the mesenchyme adjacent to rhombomeres (r)1 plus r2, r4 and r6 in three segregated streams. NCCs do not enter the intervening mesenchyme adjacent to r3 or r5, suggesting that these regions contain a NCC repulsive activity. We have used surgical manipulations in the chick to demonstrate that r3 neuroepithelium and its overlying surface ectoderm independently help maintain the NCC-free zone within r3 mesenchyme. In the absence of r3, subpopulations of NCCs enter r3 mesenchyme in a dorsolateral stream and an ectopic cranial nerve forms between the trigeminal and facial ganglia. The NCC-repulsive activity dissipates/degrades within 5-10 hours of r3 removal. Initially, r4 NCCs more readily enter the altered mesenchyme than r2 NCCs, irrespective of their maturational stage. Following surface ectoderm removal, mainly r4 NCCs enter r3 mesenchyme within 5 hours, but after 20 hours the proportions of r2 NCCs and r4 NCCs ectopically within r3 mesenchyme appear similar. PMID- 11874907 TI - Onset of the segmentation clock in the chick embryo: evidence for oscillations in the somite precursors in the primitive streak. AB - Vertebrate somitogenesis is associated with a molecular oscillator, the segmentation clock, which is defined by the periodic expression of genes related to the Notch pathway such as hairy1 and hairy2 or lunatic fringe (referred to as the cyclic genes) in the presomitic mesoderm (PSM). Whereas earlier studies describing the periodic expression of these genes have essentially focussed on later stages of somitogenesis, we have analysed the onset of the dynamic expression of these genes during chick gastrulation until formation of the first somite. We observed that the onset of the dynamic expression of the cyclic genes in chick correlated with ingression of the paraxial mesoderm territory from the epiblast into the primitive streak. Production of the paraxial mesoderm from the primitive streak is a continuous process starting with head mesoderm formation, while the streak is still extending rostrally, followed by somitic mesoderm production when the streak begins its regression. We show that head mesoderm formation is associated with only two pulses of cyclic gene expression. Because such pulses are associated with segment production at the body level, it suggests the existence of, at most, two segments in the head mesoderm. This is in marked contrast to classical models of head segmentation that propose the existence of more than five segments. Furthermore, oscillations of the cyclic genes are seen in the rostral primitive streak, which contains stem cells from which the entire paraxial mesoderm originates. This indicates that the number of oscillations experienced by somitic cells is correlated with their position along the AP axis. PMID- 11874905 TI - HEN1 functions pleiotropically in Arabidopsis development and acts in C function in the flower. AB - Four classes of floral homeotic MADS domain proteins specify the identities of the four organ types in an Arabidopsis flower. While the activities of the MADS domain proteins are essentially confined to the flower or to the inflorescence, several genes, such as APETALA2, HUA1 and HUA2, also act outside the flower in addition to their organ identity functions inside the flower. We identified a new gene, HUA ENHANCER 1 (HEN1) from a sensitized genetic screen in the hua1-1 hua2-1 background that is compromised in floral homeotic C function. We showed that HEN1, like the C function gene AGAMOUS, acts to specify reproductive organ identities and to repress A function. HEN1 also shares AG's non-homeotic function in controlling floral determinacy. HEN1 may achieve these functions by regulating the expression of AG. hen1 single mutants exhibit pleiotropic phenotypes such as reduced organ size, altered rosette leaf shape and increased number of coflorescences, during most stages of development. Therefore, HEN1, like the A function gene AP2, plays multiple roles in plant development as well as acting in organ identity specification in the flower. HEN1 codes for a novel protein and is expressed throughout the plant. PMID- 11874908 TI - Grunge, related to human Atrophin-like proteins, has multiple functions in Drosophila development. AB - We have carried out a genetic screen designed to isolate regulators of teashirt expression. One of these regulators is the Grunge gene, which encodes a protein with motifs found in human arginine-glutamic acid dipeptide repeat, Metastasis associated-like and Atrophin-1 proteins. Grunge is the only Atrophin-like protein in Drosophila, whereas several exist in humans. We provide evidence that Grunge is required for the proper regulation of teashirt but also has multiple activities in fly development. First, Grunge is crucial for correct segmentation during embryogenesis via a failure in the repression of at least four segmentation genes known to regulate teashirt. Second, Grunge acts positively to regulate teashirt expression in proximoventral parts of the leg. Grunge has other regulatory functions in the leg, including the patterning of ventral parts along the entire proximodistal axis and the proper spacing of bristles in all regions. PMID- 11874909 TI - MAX1 and MAX2 control shoot lateral branching in Arabidopsis. AB - Plant shoots elaborate their adult form by selective control over the growth of both their primary shoot apical meristem and their axillary shoot meristems. We describe recessive mutations at two loci in Arabidopsis, MAX1 and MAX2, that affect the selective repression of axillary shoots. All the first order (but not higher order) axillary shoots initiated by mutant plants remain active, resulting in bushier shoots than those of wild type. In vegetative plants where axillary shoots develop in a basal to apical sequence, the mutations do not clearly alter node distance, from the shoot apex, at which axillary shoot meristems initiate but shorten the distance at which the first axillary leaf primordium is produced by the axillary shoot meristem. A small number of mutant axillary shoot meristems is enlarged and, later in development, a low proportion of mutant lateral shoots is fasciated. Together, this suggests that MAX1 and MAX2 do not control the timing of axillary meristem initiation but repress primordia formation by the axillary meristem. In addition to shoot branching, mutations at both loci affect leaf shape. The mutations at MAX2 cause increased hypocotyl and petiole elongation in light-grown seedlings. Positional cloning identifies MAX2 as a member of the F-box leucine-rich repeat family of proteins. MAX2 is identical to ORE9, a proposed regulator of leaf senescence ( Woo, H. R., Chung, K. M., Park, J.-H., Oh, S. A., Ahn, T., Hong, S. H., Jang, S. K. and Nam, H. G. (2001) Plant Cell 13, 1779-1790). Our results suggest that selective repression of axillary shoots involves ubiquitin-mediated degradation of as yet unidentified proteins that activate axillary growth. PMID- 11874910 TI - Development of pigment-cup eyes in the polychaete Platynereis dumerilii and evolutionary conservation of larval eyes in Bilateria. AB - The role of Pax6 in eye development in insects and vertebrates supports the view that their eyes evolved from simple pigment-cup ocelli present in their last common ancestors (Urbilateria). The cerebral eyes in errant polychaetes represent prototype invertebrate pigment-cup ocelli and thus resemble the presumed ancestral eyes. We have analysed expression of conserved eye specification genes in the early development of larval and adult pigment-cup eyes in Platynereis dumerilii (Polychaeta, Annelida, Lophotrochozoa). Both larval and adult eyes form in close vicinity of the optic anlagen on both sides of the developing brain ganglia. While pax6 is expressed in the larval, but not in the developing, adult eyes, expression of six1/2 from trochophora stages onwards specifically outlines the optic anlagen and thus covers both the developing larval and adult eyes. Using Platynereis rhabdomeric opsin as differentiation marker, we show that the first pair of adult eye photoreceptor cells is detected within bilateral clusters that transitorily express ath, the Platynereis atonal orthologue, thus resembling proneural sensory clusters. Our data indicate that--similar to insects, but different from the vertebrates--polychaete six1/2 expression outlines the entire visual system from early developmental stages onwards and ath-positive clusters generate the first photoreceptor cells to appear. We propose that pax6-, six1/2- and ath-positive larval eyes, as found in today's trochophora, were present already in Urbilateria. PMID- 11874911 TI - Sexually dimorphic development of mouse primordial germ cells: switching from oogenesis to spermatogenesis. AB - During embryogenesis, primordial germ cells (PGCs) have the potential to enter either spermatogenesis or oogenesis. In a female genital ridge, or in a non gonadal environment, PGCs develop as meiotic oocytes. However, male gonadal somatic cells inhibit PGCs from entering meiosis and direct them to a spermatogenic fate. We have examined the ability of PGCs from male and female embryos to respond to the masculinising environment of the male genital ridge, defining a temporal window during which PGCs retain a bipotential fate. To help understand how PGCs respond to the male gonadal environment, we have identified molecular differences between male PGCs that are committed to spermatogenesis and bipotential female PGCs. Our results suggest that one way in which PGCs respond to this masculinising environment is to synthesise prostaglandin D(2). We show that this signalling molecule can partially masculinise female embryonic gonads in culture, probably by inducing female supporting cells to differentiate into Sertoli cells. In the developing testis, prostaglandin D(2) may act as a paracrine factor to induce Sertoli cell differentiation. Thus part of the response of PGCs to the male gonadal environment is to generate a masculinising feedback loop to ensure male differentiation of the surrounding gonadal somatic cells. PMID- 11874913 TI - her1 and the notch pathway function within the oscillator mechanism that regulates zebrafish somitogenesis. AB - Somite formation is thought to be regulated by an unknown oscillator mechanism that causes the cells of the presomitic mesoderm to activate and then repress the transcription of specific genes in a cyclical fashion. These oscillations create stripes/waves of gene expression that repeatedly pass through the presomitic mesoderm in a posterior-to-anterior direction. In both the mouse and the zebrafish, it has been shown that the notch pathway is required to create the stripes/waves of gene expression. However, it is not clear if the notch pathway comprises part of the oscillator mechanism or if the notch pathway simply coordinates the activity of the oscillator among neighboring cells. In the zebrafish, oscillations in the expression of a hairy-related transcription factor, her1 and the notch ligand deltaC precede somite formation. Our study focuses on how the oscillations in the expression of these two genes is affected in the mutants aei/deltaD and des/notch1, in 'morpholino knockdowns' of deltaC and her1 and in double 'mutant' combinations. This analysis indicates that these oscillations in gene expression are created by a genetic circuit comprised of the notch pathway and the notch target gene her1. We also show that a later function of the notch pathway can create a segmental pattern even in the absence of prior oscillations in her1 and deltaC expression. PMID- 11874912 TI - The Sox-domain containing gene Dichaete/fish-hook acts in concert with vnd and ind to regulate cell fate in the Drosophila neuroectoderm. AB - In the Drosophila embryonic central nervous system, neural stem cells, called neuroblasts, acquire fates in a position-specific manner. Recent work has identified a set of genes that functions along the dorsoventral axis to enable neuroblasts that develop in different dorsoventral domains to acquire distinct fates. These genes include the evolutionarily conserved transcription factors ventral nerve cord defective and intermediate neuroblasts defective, as well as the Drosophila EGF receptor. We show that the Sox-domain-containing gene Dichaete/fish-hook also plays a crucial role to pattern the neuroectoderm along the DV axis. Dichaete is expressed in the medial and intermediate columns of the neuroectoderm, and mutant analysis indicates that Dichaete regulates cell fate and neuroblast formation in these domains. Molecular epistasis tests, double mutant analysis and dosage-sensitive interactions demonstrate that during these processes, Dichaete functions in parallel with ventral nerve cord defective and intermediate neuroblasts defective, and downstream of EGF receptor signaling to mediate its effect on development. These results identify Dichaete as an important regulator of dorsoventral pattern in the neuroectoderm, and indicate that Dichaete acts in concert with ventral nerve cord defective and intermediate neuroblasts defective to regulate pattern and cell fate in the neuroectoderm. PMID- 11874914 TI - A novel thioredoxin-like protein encoded by the C. elegans dpy-11 gene is required for body and sensory organ morphogenesis. AB - Sensory ray morphogenesis in C. elegans requires active cellular interaction regulated by multiple genetic activities. We report here the cloning of one of these genes, dpy-11, which encodes a membrane-associated thioredoxin-like protein. The DPY-11 protein is made exclusively in the hypodermis and resides in the cytoplasmic compartment. Whereas the TRX domain of DPY-11 displays a catalytic activity in vitro, mapping of lesions in different mutant alleles and functional analysis of deletion transgenes reveal that both this enzymatic activity and transmembrane topology are essential for determining body shape and ray morphology. Based on the abnormal features in both the expressing and non expressing ray cells, we propose that the DPY-11 is required in the hypodermis for modification of its substrates. In turn, ray cell interaction and the whole morphogenetic process can be modulated by these substrate molecules. PMID- 11874915 TI - Short Stop provides an essential link between F-actin and microtubules during axon extension. AB - Coordination of F-actin and microtubule dynamics is important for cellular motility and morphogenesis, but little is known about underlying mechanisms. short stop (shot) encodes an evolutionarily conserved, neuronally expressed family of rod-like proteins required for sensory and motor axon extension in Drosophila melanogaster. We identify Shot isoforms that contain N-terminal F actin and C-terminal microtubule-binding domains, and that crosslink F-actin and microtubules in cultured cells. The F-actin- and microtubule-binding domains of Shot are required in the same molecule for axon extension, though the length of the connecting rod domain can be dramatically reduced without affecting activity. Shot therefore functions as a cytoskeletal crosslinker in axon extension, rather than mediating independent interactions with F-actin and microtubules. A Ca(2+) binding motif located adjacent to the microtubule-binding domain is also required for axon extension, suggesting that intracellular Ca(2+) release may regulate Shot activity. These results suggest that Shot coordinates regulated interactions between F-actin and microtubules that are crucial for neuronal morphogenesis. PMID- 11874916 TI - Novel conserved elements upstream of the H19 gene are transcribed and act as mesodermal enhancers. AB - The reciprocally imprinted H19 and Igf2 genes form a co-ordinately regulated 130 kb unit in the mouse controlled by widely dispersed enhancers, epigenetically modified silencers and an imprinting control region (ICR). Comparative human and mouse genomic sequencing between H19 and Igf2 revealed two novel regions of strong homology upstream of the ICR termed H19 upstream conserved regions (HUCs). Mouse HUC1 and HUC2 act as potent enhancers capable of driving expression of an H19 reporter gene in a range of mesodermal tissues. Intriguingly, the HUC sequences are also transcribed bi-allelically in mouse and human, but their expression pattern in neural and endodermal tissues in day 13.5 embryos is distinct from their enhancer function. The location of the HUC mesodermal enhancers upstream of the ICR and H19, and their capacity for interaction with both H19 and Igf2 requires critical re-evaluation of the cis-regulation of imprinted gene expression of H19 and Igf2 in a range of mesodermal tissues. We propose that these novel sequences interact with the ICR at H19 and the epigenetically regulated silencer at differentially methylated region 1 (DMR1) of Igf2. PMID- 11874917 TI - Drosophila myosin phosphatase and its role in dorsal closure. AB - Myosin phosphatase negatively regulates nonmuscle myosin II through dephosphorylation of the myosin regulatory light chain (MRLC). Its regulatory myosin-binding subunit, MBS, is responsible for regulating the catalytic subunit in response to upstream signals and for determining the substrate specificity. DMBS, the Drosophila homolog of MBS, was identified to study the roles of myosin phosphatase in morphogenesis. The embryos defective for both maternal and zygotic DMBS demonstrated a failure in dorsal closure. In the mutant embryos, the defects were mainly confined to the leading edge cells which failed to fully elongate. Ectopic accumulation of phosphorylated MRLC was detected in lateral region of the leading edge cells, suggesting that the role of DMBS is to repress the activation of nonmuscle myosin II at the subcellular location for coordinated cell shape change. Aberrant accumulation of F-actin within the leading edge cells may correspond to the morphological aberrations of such cells. Similar defects were seen in embryos overexpressing Rho-kinase, suggesting that myosin phosphatase and Rho-kinase function antagonistically. The genetic interaction of DMBS with mutations in the components of the Rho signaling cascade also indicates that DMBS functions antagonistically to the Rho signal transduction pathway. The results indicate an important role for myosin phosphatase in morphogenesis. PMID- 11874918 TI - Exploring the myriapod body plan: expression patterns of the ten Hox genes in a centipede. AB - The diversity of the arthropod body plan has long been a fascinating subject of study. A flurry of recent research has analyzed Hox gene expression in various arthropod groups, with hopes of gaining insight into the mechanisms that underlie their evolution. The Hox genes have been analyzed in insects, crustaceans and chelicerates. However, the expression patterns of the Hox genes have not yet been comprehensively analyzed in a myriapod. We present the expression patterns of the ten Hox genes in a centipede, Lithobius atkinsoni, and compare our results to those from studies in other arthropods. We have three major findings. First, we find that Hox gene expression is remarkably dynamic across the arthropods. The expression patterns of the Hox genes in the centipede are in many cases intermediate between those of the chelicerates and those of the insects and crustaceans, consistent with the proposed intermediate phylogenetic position of the Myriapoda. Second, we found two 'extra' Hox genes in the centipede compared with those in Drosophila. Based on its pattern of expression, Hox3 appears to have a typical Hox-like role in the centipede, suggesting that the novel functions of the Hox3 homologs zen and bicoid were adopted somewhere in the crustacean-insect clade. In the centipede, the expression of the gene fushi tarazu suggests that it has both a Hox-like role (as in the mite), as well as a role in segmentation (as in insects). This suggests that this dramatic change in function was achieved via a multifunctional intermediate, a condition maintained in the centipede. Last, we found that Hox expression correlates with tagmatic boundaries, consistent with the theory that changes in Hox genes had a major role in evolution of the arthropod body plan. PMID- 11874919 TI - Parasegmental organization of the spider embryo implies that the parasegment is an evolutionary conserved entity in arthropod embryogenesis. AB - Spiders belong to the chelicerates, which is a basal arthropod group. To shed more light on the evolution of the segmentation process, orthologs of the Drosophila segment polarity genes engrailed, wingless/Wnt and cubitus interruptus have been recovered from the spider Cupiennius salei. The spider has two engrailed genes. The expression of Cs-engrailed-1 is reminiscent of engrailed expression in insects and crustaceans, suggesting that this gene is regulated in a similar way. This is different for the second spider engrailed gene, Cs engrailed-2, which is expressed at the posterior cap of the embryo from which stripes split off, suggesting a different mode of regulation. Nevertheless, the Cs-engrailed-2 stripes eventually define the same border as the Cs-engrailed-1 stripes. The spider wingless/Wnt genes are expressed in different patterns from their orthologs in insects and crustaceans. The Cs-wingless gene is expressed in iterated stripes just anterior to the engrailed stripes, but is not expressed in the most ventral region of the germ band. However, Cs-Wnt5-1 appears to act in this ventral region. Cs-wingless and Cs-Wnt5-1 together seem to perform the role of insect wingless. Although there are differences, the wingless/Wnt-expressing cells and en-expressing cells seem to define an important boundary that is conserved among arthropods. This boundary may match the parasegmental compartment boundary and is even visible morphologically in the spider embryo. An additional piece of evidence for a parasegmental organization comes from the expression domains of the Hox genes that are confined to the boundaries, as molecularly defined by the engrailed and wingless/Wnt genes. Parasegments, therefore, are presumably important functional units and conserved entities in arthropod development and form an ancestral character of arthropods. The lack of by engrailed and wingless/Wnt-defined boundaries in other segmented phyla does not support a common origin of segmentation. PMID- 11874920 TI - Sensory neurons of the Atonal lineage pioneer the formation of glomeruli within the adult Drosophila olfactory lobe. AB - The first centers for processing of odor information by animals lie in the olfactory lobe. Sensory neurons from the periphery synapse with interneurons in anatomically recognizable units, termed glomeruli, seen in both insects and vertebrates. The mechanisms that underlie the formation of functional maps of the odor-world in the glomeruli within the olfactory lobe remains unclear. We address the basis of sensory targeting in the fruitfly Drosophila and show that one class of sensory neurons, those of the Atonal lineage, plays a crucial role in glomerular patterning. Atonal-dependent neurons pioneer the segregation of other classes of sensory neurons into distinct glomeruli. Furthermore, correct sensory innervation is necessary for the arborization of projection neurons into glomeruli and for the elaboration of processes of central glial cells into the lobe. PMID- 11874921 TI - Identification of a developmental transition in plasmodesmatal function during embryogenesis in Arabidopsis thaliana. AB - Plasmodesmata provide routes for communication and nutrient transfer between plant cells by interconnecting the cytoplasm of adjacent cells. A simple fluorescent tracer loading assay was developed to monitor patterns of cell-to cell transport via plasmodesmata specifically during embryogenesis. A developmental transition in plasmodesmatal size exclusion limit was found to occur at the torpedo stage of embryogenesis in Arabidopsis; at this time, plasmodesmata are down-regulated, allowing transport of small (approx. 0.5 kDa) but not large (approx. 10 kDa) tracers. This assay system was used to screen for embryo-defective mutants, designated increased size exclusion limit of plasmodesmata (ise), that maintain dilated plasmodesmata at the torpedo stage. The morphology of ise1 and ise2 mutants discussed here resembled that of the wild type during embryo development, although the rate of their embryogenesis was slower. The ISE1 gene was mapped to position 13 cM on chromosome I using PCR based biallelic markers. ise2 was found to be allelic to the previously characterized mutant emb25 which maps to position 100 cM on chromosome I. The results presented have implications for intercellular signaling pathways that regulate embryonic development, and furthermore represent the first attempt to screen directly for mutants of Arabidopsis with altered size exclusion limit of plasmodesmata. PMID- 11874922 TI - Control of Drosophila imaginal disc development by rotund and roughened eye: differentially expressed transcripts of the same gene encoding functionally distinct zinc finger proteins. AB - The Drosophila rotund gene is required in the wings, antenna, haltere, proboscis and legs. A member of the Rac family of GTPases, denoted the rotund racGAP gene, was previously identified in the rotund region. However, previous studies indicated that rotund racGAP was not responsible for the rotund phenotypes and that the rotund gene had yet to be identified. We have isolated the rotund gene and show that it is a member of the Kruppel family of zinc finger genes. The adjacent roughened eye locus specifically affects the eye and is genetically separable from rotund. However, roughened eye and rotund are tightly linked, and we have therefore also isolated the roughened eye transcript. Intriguingly, we show that roughened eye is part of the rotund gene but is represented by a different transcript. The rotund and roughened eye transcripts result from the utilization of two different promoters that direct expression in non-overlapping domains in the larval imaginal discs. The predicted Rotund and Roughened Eye proteins share the same C-terminal region, including the zinc finger domain, but differ in their N-terminal regions. Each cDNA can rescue only the corresponding mutation and show negative effects when expressed in each others domain of expression. These results indicate that in addition to the differential expression of rotund and roughened eye, their proteins have distinct activities. rotund and roughened eye act downstream of early patterning genes such as dachshund and appear to be involved in Notch signaling by regulating Delta, scabrous and SERRATE: PMID- 11874923 TI - The Kallmann syndrome gene homolog in C. elegans is involved in epidermal morphogenesis and neurite branching. AB - Kallmann syndrome is an inherited disorder defined by the association of anosmia and hypogonadism, owing to impaired targeting and migration of olfactory axons and gonadotropin-releasing hormone secreting neurons. The gene responsible for the X-linked form of Kallmann syndrome, KAL-1, encodes a secreted protein of still elusive function. It has been proposed that KAL-1 might be involved in some aspects of olfactory axon guidance. However, the unavailability of a mouse model, and the difficulties in studying cellular and axonal migration in vertebrates have hampered an understanding of its function. We have identified the C. elegans homolog, kal-1, and document its function in vivo. We show that kal-1 is part of a mechanism by which neurons influence migration and adhesion of epidermal cells undergoing morphogenesis during ventral enclosure and male tail formation. We also show that kal-1 affects neurite outgrowth in vivo by modulating branching. Finally, we find that human KAL-1 cDNA can compensate for the loss of worm kal-1 and that overexpression of worm or human KAL-1 cDNAs in the nematode results in the same phenotypes. These data indicate functional conservation between the human and nematode proteins and establish C. elegans as a powerful animal in which to investigate KAL function in vivo. Our findings add a new player to the set of molecules, which appear to underlie both morphogenesis and axonal/neuronal navigation in vertebrates and invertebrates. PMID- 11874924 TI - Dietary fat and meat intake in relation to risk of type 2 diabetes in men. AB - OBJECTIVE: To examine dietary fat and meat intake in relation to risk of type 2 diabetes. RESEARCH DESIGN AND METHODS: We prospectively followed 42,504 male participants of the Health Professionals Follow-Up Study who were aged 40-75 years and free of diagnosed diabetes, cardiovascular disease, and cancer in 1986. Diet was assessed by a validated food frequency questionnaire and updated in 1990 and 1994. During 12 years of follow-up, we ascertained 1,321 incident cases of type 2 diabetes. RESULTS: Intakes of total fat (multivariate RR for extreme quintiles 1.27, CI 1.04-1.55, P for trend=0.02) and saturated fat (1.34, 1.09 1.66, P for trend=0.01) were associated with a higher risk of type 2 diabetes. However, these associations disappeared after additional adjustment for BMI (total fat RR 0.97, CI 0.79-1.18; saturated fat 0.97, 0.79-1.20). Intakes of oleic acid, trans-fat, long-chain n-3 fat, and alpha-linolenic acid were not associated with diabetes risk after multivariate adjustment. Linoleic acid was associated with a lower risk of type 2 diabetes in men <65 years of age (RR 0.74, CI 0.60-0.92, P for trend=0.01) and in men with a BMI <25 kg/m(2) (0.53, 0.33 0.85, P for trend=0.006) but not in older and obese men. Frequent consumption of processed meat was associated with a higher risk for type 2 diabetes (RR 1.46, CI 1.14-1.86 for > or = 5/week vs. <1/month, P for trend <0.0001). CONCLUSIONS: Total and saturated fat intake were associated with a higher risk of type 2 diabetes, but these associations were not independent of BMI. Frequent consumption of processed meats may increase risk of type 2 diabetes. PMID- 11874925 TI - Effect of a high-protein, high-monounsaturated fat weight loss diet on glycemic control and lipid levels in type 2 diabetes. AB - OBJECTIVE: To determine the effect of a high-protein (HP) weight loss diet compared with a lower-protein (LP) diet on fat and lean tissue and fasting and postprandial glucose and insulin concentrations. RESEARCH DESIGN AND METHODS: Replacing dietary protein for carbohydrate (CHO) during energy restriction and weight loss has been effective in sparing lean mass and improving insulin sensitivity in obese subjects but has not been tested in subjects with type 2 diabetes. We compared an HP diet (28% protein, 42% CHO, 28% fat [8% saturated fatty acids, 12% monounsaturated fatty acids, 5% polyunsaturated fatty acids]) with an LP diet (16% protein, 55% CHO, 26% fat [8% saturated fatty acids, 11% monounsaturated fatty acids, 5% polyunsaturated fatty acids]) in 54 obese men and women with type 2 diabetes during 8 weeks of energy restriction (1,600 kcal) and 4 weeks of energy balance. Body composition was determined by dual-energy X-ray absorptiometry at weeks 0 and 12. RESULTS: Overall, weight loss of 5.2 +/- 1.8 kg was achieved independently of diet composition. However, women on the HP diet lost significantly more total (5.3 vs. 2.8 kg, P=0.009) and abdominal (1.3 vs. 0.7 kg, P=0.006) fat compared with the women on the LP diet, whereas, in men, there was no difference in fat loss between diets (3.9 vs. 5.1 kg). Total lean mass decreased in all subjects independently of diet composition. LDL cholesterol reduction was significantly greater on the HP diet (5.7%) than on the LP diet (2.7%) (P < 0.01). CONCLUSIONS: Both dietary patterns resulted in improvements in the cardiovascular disease (CVD) risk profile as a consequence of weight loss. However, the greater reductions in total and abdominal fat mass in women and greater LDL cholesterol reduction observed in both sexes on the HP diet suggest that it is a valid diet choice for reducing CVD risk in type 2 diabetes. PMID- 11874926 TI - Effects of an energy-restrictive diet with or without exercise on abdominal fat, intermuscular fat, and metabolic risk factors in obese women. AB - OBJECTIVE: The primary objective was to examine whether the combination of diet and aerobic exercise (DA) or diet and resistance exercise (DR) is associated with greater improvements in metabolic risk factors by comparison to diet only (DO) in obese women. A second objective considered whether reductions in metabolic risk factors are related to concurrent changes in abdominal and/or intermuscular fat distribution. RESEARCH DESIGN AND METHODS: A total of 38 premenopausal obese women were randomly assigned to one of three 16-week treatments: DO (n=13), DA (n=11), or DR (n=14). Plasma glucose, insulin, and lipid levels were measured in a fasting state and after a 75-g oral glucose challenge (oral glucose tolerance test [OGTT]). Total, abdominal subcutaneous, visceral, and intermuscular fat were measured by magnetic resonance imaging. RESULTS: Significant reductions (P < 0.02) in body weight (approximately 10 kg or 10%) and in total, abdominal subcutaneous, visceral, and intermuscular fat were observed within each group. Fasting and OGTT insulin, total cholesterol, LDL cholesterol, and apolipoprotein B also decreased within each group (P < or = 0.02). The changes in the body fat and metabolic variables were not different across treatment (P > 0.05). Visceral fat alone was related to the metabolic risk factors both before and after the treatment. CONCLUSIONS: Weight loss was associated with reductions in metabolic risk factors in obese women. The improvement in the metabolic profile was not enhanced by the addition of aerobic or resistance exercise. The findings reinforce the importance of diminished visceral fat in the treatment of insulin resistance. PMID- 11874927 TI - Comparison of insulin aspart with buffered regular insulin and insulin lispro in continuous subcutaneous insulin infusion: a randomized study in type 1 diabetes. AB - OBJECTIVE: To compare the safety and efficacy of insulin aspart (IAsp), buffered regular insulin (BR), and insulin lispro administered by continuous subcutaneous insulin infusion (CSII) in patients with type 1 diabetes. RESEARCH DESIGN AND METHODS: After completing a 4-week run-in period with BR, 146 adult patients with type 1 diabetes (with pretrial CSII experience) were randomly assigned (2:2:1) to CSII treatment with IAsp, BR, or lispro for 16 weeks in a multicenter, open label, randomized, parallel-group study. Bolus insulin doses were administered 30 min before meals (BR) or immediately before meals (IAsp or lispro). RESULTS: Treatment groups had similar baseline HbA(1c) (7.3% +/- 0.7 for IAsp, 7.5% +/- 0.8 for BR, and 7.3% +/- 0.7 for lispro). After 16 weeks of treatment, HbA1c values were relatively unchanged from baseline, and the mean changes in baseline HbA1c values were not significantly different between the three groups (0.00 +/- 0.51, 0.15 +/- 0.63, and 0.18 +/- 0.84 for the IAsp, BR, and lispro groups, respectively). The rates of hypoglycemic episodes (blood glucose <50 mg/dl) per patient per month were similar (3.7, 4.8, and 4.4 for the IAsp, BR, and lispro groups, respectively). Clogs/blockages in pumps or infusion sets were infrequent; most subjects (76, 83, and 75% in the IAsp, BR, and lispro groups, respectively) had < or = 1 clog or blockage per 4 weeks during the trial. CONCLUSIONS: Insulin aspart in CSII was as efficacious and well tolerated as BR and lispro and is a suitable insulin for continuous subcutaneous insulin infusion using external pumps. PMID- 11874928 TI - Intensive lifestyle changes are necessary to improve insulin sensitivity: a randomized controlled trial. AB - OBJECTIVE: The extent to which lifestyle must be altered to improve insulin sensitivity has not been established. This study compares the effect on insulin sensitivity of current dietary and exercise recommendations with a more intensive intervention in normoglycemic insulin-resistant individuals. RESEARCH DESIGN AND METHODS: Seventy-nine normoglycemic insulin-resistant (determined by the euglycemic insulin clamp) men and women were randomized to either a control group or one of two combined dietary and exercise programs. One group (modest level) was based on current recommendations and the other on a more intensive dietary and exercise program. Insulin sensitivity was measured using a euglycemic insulin clamp, body composition was measured using dual-energy X-ray absorptiometry, and anthropometry and aerobic fitness were assessed before and after a 4-month intervention period. Four-day dietary intakes were recorded, and fasting glucose, insulin, and lipids were measured. RESULTS: Only the intensive group showed a significant improvement in insulin sensitivity (23% increase, P=0.006 vs. 9% in the modest group, P=0.23). This was associated with a significant improvement in aerobic fitness (11% increase in the intensive group, P=0.02 vs. 1% in the modest group, P=0.94) and a greater fiber intake, but no difference in reported total or saturated dietary fat. CONCLUSIONS: Current clinical dietary and exercise recommendations, even when vigorously implemented, did not significantly improve insulin sensitivity; however, a more intensive program did. Improved aerobic fitness appeared to be the major difference between the two intervention groups, although weight loss and diet composition may have also played an important role in determining insulin sensitivity. PMID- 11874929 TI - Basal and postglucagon C-peptide levels in Ethiopians with diabetes. AB - OBJECTIVE: To study basal C-peptide (BCP) and postglucagon C-peptide (PGCP) levels in Ethiopians with diabetes. RESEARCH DESIGN AND METHODS: A total of 56 subjects with type 1 diabetes, 97 subjects with type 2 diabetes, and 50 control subjects were recruited from a hospital in Ethiopia. BCP was determined in all subjects and PGCP in 86 subjects. RESULTS: Mean (+/- SEM) BCP, PGCP, and the increment after glucagon in type 1 diabetic subjects (0.14 +/- 0.04, 0.22 +/- 0.11, and 0.08 +/- 0.05 nmol/l, respectively) were lower (P < 0.001) than those in type 2 diabetic subjects (0.66 +/- 0.04, 1.25 +/- 0.10, and 0.56 +/- 0.06 nmol/l, respectively) or control subjects (0.54 +/- 0.04, 1.52 +/- 0.26, and 1.11 +/- 0.24 nmol/l, respectively). The mean BCP level was higher in type 2 diabetic subjects than control subjects (P=0.015), whereas the mean increment was lower (P=0.005). Insulin-treated type 2 diabetic subjects, compared with non-insulin treated type 2 diabetic subjects, had lower mean BCP (0.55 +/- 0.08 nmol/l [n=37] vs. 0.73 +/- 0.04 [n=60], P=0.001), lower PGCP (0.97 +/- 0.20 nmol/l [n=18] vs. 1.40 +/- 0.11 [n=35], P=0.010), and a lower C-peptide increment (0.34 +/- 0.06 [n=18] vs. 0.67 +/- 0.07 nmol/l [n=35], P=0.003). In both the type 1 and type 2 diabetic groups, those with BCP levels <0.2 nmol/l had lower BMI than those with higher BCP levels (P=0.023 and P < 0.001, respectively). CONCLUSIONS: Combined with clinical criteria, C-peptide levels are good discriminators between type 1 and type 2 diabetes in Ethiopians and may also be useful in identifying subjects with type 2 diabetes who require insulin therapy. There is a subgroup of type 2 diabetic subjects with features of type 1 diabetes. PMID- 11874930 TI - Health-related quality of life and treatment satisfaction in Dutch patients with type 2 diabetes. AB - OBJECTIVE: To estimate the health-related quality of life (HRQOL) and treatment satisfaction for patients with type 2 diabetes in the Netherlands and to examine which patient characteristics are associated with quality of life and treatment satisfaction. RESEARCH DESIGN AND METHODS: For a sample of 1,348 type 2 diabetes patients, recruited by 29 general practitioners, we collected data regarding HRQOL. This study was performed as part of a larger European study (Cost of Diabetes in Europe - Type 2 [CODE-2]). We used a generic instrument (Euroqol 5D) to measure HRQOL. Treatment satisfaction was assessed using the Diabetes Treatment Satisfaction Questionnaire. RESULTS: Patients without complications had an HRQOL (0.74) only slightly lower than similarly aged persons in the general population. Insulin therapy, obesity, and complications were associated with a lower HRQOL, independent of age and sex. Although higher fasting blood glucose and HbA1c levels were negatively associated with HRQOL, these factors were not significant after adjustment for other factors using multivariate analysis. Overall treatment satisfaction was very high. Younger patients, patients using insulin, and patients with higher HbA1c levels were less satisfied with the treatment than other patients. CONCLUSIONS: Obesity and the presence of complications are important determinants of HRQOL in patients with type 2 diabetes. PMID- 11874931 TI - Comorbid depression is associated with increased health care use and expenditures in individuals with diabetes. AB - OBJECTIVE: This study ascertained the odds of diagnosed depression in individuals with diabetes and the relation between depression and health care use and expenditures. RESEARCH DESIGN AND METHODS: First, we compared data from 825 adults with diabetes with that from 20,688 adults without diabetes using the 1996 Medical Expenditure Panel Survey (MEPS). Second, in patients with diabetes, we compared depressed and nondepressed individuals to identify differences in health care use and expenditures. Third, we adjusted use and expenditure estimates for differences in age, sex, race/ethnicity, health insurance, and comorbidity with analysis of covariance. Finally, we used the Consumer Price Index to adjust expenditures for inflation and used SAS and SUDAAN software for statistical analyses. RESULTS: Individuals with diabetes were twice as likely as a comparable sample from the general U.S. population to have diagnosed depression (odds ratio 1.9, 95% CI 1.5-2.5). Younger adults (<65 years), women, and unmarried individuals with diabetes were more likely to have depression. Patients with diabetes and depression had higher ambulatory care use (12 vs. 7, P < 0.0001) and filled more prescriptions (43 vs. 21, P < 0.0001) than their counterparts without depression. Finally, among individuals with diabetes, total health care expenditures for individuals with depression was 4.5 times higher than that for individuals without depression ($247,000,000 vs. $55,000,000, P < 0.0001). CONCLUSIONS: The odds of depression are higher in individuals with diabetes than in those without diabetes. Depression in individuals with diabetes is associated with increased health care use and expenditures, even after adjusting for differences in age, sex, race/ethnicity, health insurance, and comorbidity. PMID- 11874932 TI - Diabetes-related morbidity and mortality in a national sample of U.S. elders. AB - OBJECTIVE: Although the number of elders with diabetes has increased dramatically, there are few data on rates of mortality and serious complications in older populations with diabetes. To determine such rates, we conducted a population-based, nonconcurrent cohort study using claims data from the 1994-1996 Medicare 5% Standard Analytical File. RESEARCH DESIGN AND METHODS: Codes from the ICD-9 were used to identify diabetes and the following complications: amputation, lower extremity infection, gangrene, blindness, acute myocardial infarction, ischemic heart disease, stroke, and metabolic disorders. Using these codes, we assembled a cohort of 148,562 Medicare Part A and B beneficiaries who were > or = 65 years of age, who were alive on 1 January 1995, who were not in managed care in 1994, and who had a diabetes-related claim in 1994. Age-specific rates of death and complications were then calculated. RESULTS: During 24 months of follow up, 22,044 (14.8%) elders with diabetes died. Death rates in men and women increased significantly with age. Compared with their counterparts in the general U.S. population, elders with diabetes suffered excess mortality at every age group, corresponding to an overall standardized mortality ratio of 1.41 (95% CI 1.39,1.43). The incidence of ischemic heart disease and stroke was 181.5 and 126.2 per 1,000 person-years, respectively, which was higher than the incidence of all other diabetes-related complications. CONCLUSIONS: In every age group, elders with diabetes have significantly higher all-cause mortality rates than the general population. Medicare data may be useful in monitoring trends in diabetes related morbidity and total mortality in U.S. elders with diabetes. PMID- 11874933 TI - Lifetime costs of complications resulting from type 2 diabetes in the U.S. AB - OBJECTIVE: To model the lifetime costs associated with complications of type 2 diabetes. RESEARCH DESIGN AND METHODS: A cohort of 10,000 patients with diabetes was simulated using a model based on existing epidemiological studies. Complication rates were estimated for various stages of macrovascular disease, nephropathy, retinopathy, neuropathy, and hypoglycemia. At the beginning of the simulation, patients were assumed to have been treated for 5 years and have a mean HbA1c of 8.4. From the U.K. Prospective Diabetes Study, it was estimated that on current therapies, the HbA1c would drift upward on average 0.15% per year. Direct medical costs of managing each complication were estimated (in 2000 U.S. dollars) from all-payor databases, surveys, and literature. RESULTS: Macrovascular disease was estimated to be the largest cost component, accounting for 85% of cumulative costs of complications over the first 5 years. The costs of complications were estimated to be $47,240 per patient over 30 years, on average. The management of macrovascular disease is estimated to be the largest cost component, accounting for 52% of the costs; nephropathy accounts for 21%, neuropathy accounts for 17%, and retinopathy accounts for 10% of the costs of complications. CONCLUSIONS: The complications of diabetes account for substantial costs, with management of macrovascular disease being the largest and earliest. If improving glycemic control prevents complications, it will reduce these costs. PMID- 11874934 TI - The impact of cardiovascular disease on medical care costs in subjects with and without type 2 diabetes. AB - OBJECTIVE: We examined whether cardiovascular disease (CVD) affects medical care costs differently in subjects with and without diabetes and explored the impact of CVD on costs across the dimensions of age and diabetes duration. RESEARCH DESIGN AND METHODS: We compared the prevalence of CVD and medical care costs for subjects with and without CVD in all 16,180 full-year health maintenance organization members in 1999 who had been diagnosed with type 2 diabetes and in control members matched by year of birth and sex. We ascertained diagnoses from the Kaiser Permanente Northwest Region's electronic ambulatory medical record and from hospital discharge datafiles. Utilization from these and other data systems were multiplied by unit costs. RESULTS: CVD was 76% more prevalent in subjects with diabetes, but the risk ratios of more severe forms of CVD were even greater. Risk ratios for CVD were greatest in younger subjects. Cost profiles for subjects with both CVD and diabetes differed markedly from those with diabetes but without CVD. In the latter group, costs grew steadily with age, whereas in the former group, costs peaked in the 55- to 64-year age group before declining with age. CONCLUSIONS: The types of CVD present in diabetic patients are more likely to be more severe and therefore more costly than in similar subjects without diabetes. CVD also disproportionately affects younger diabetic subjects. Finally, when CVD is present in diabetes, more costs occur earlier in life as well as earlier in the course of diabetes. PMID- 11874935 TI - A comparison of rates, risk factors, and outcomes of gestational diabetes between aboriginal and non-aboriginal women in the Saskatoon health district. AB - OBJECTIVE: To determine possible differences in gestational diabetes mellitus (GDM) between aboriginal and non-aboriginal people in the Saskatoon Health District. RESEARCH DESIGN AND METHODS: This was a prospective survey of all women admitted for childbirth to the Saskatoon Royal University Hospital between January and July 1998. We compared prevalence rates, risk factors, and outcomes of GDM between aboriginal and non-aboriginal women. RESULTS: Information was obtained from 2,006 women, of whom 252 aboriginal and 1,360 non-aboriginal subjects had been tested for GDM. The overall rates of GDM were 3.5% for women in the general population and 11.5% for aboriginal women. For those living within the Saskatoon Health District, GDM rates were 3.7 and 6.4%, respectively. Multivariate analysis demonstrated that aboriginal ethnicity, most notably when combined with obesity, was an independent predictor for GDM. Pregravid BMI > or = 27 kg/m(2) and maternal age > or = 33 years were the most important risk factors for GDM in aboriginal women, whereas previous GDM, family history of diabetes, and maternal age > or = 38 years were the strongest predictors for GDM in non aboriginal women. CONCLUSIONS: There may be fundamental differences in GDM between aboriginal and non-aboriginal people. Because GDM contributes to an increased risk for type 2 diabetes in aboriginal women and their offspring, the impact of prevention and optimal treatment of GDM on the type 2 diabetes epidemic in susceptible populations are important areas for further investigation. PMID- 11874936 TI - Type 2 diabetes and metabolic syndrome in Filipina-American women : a high-risk nonobese population. AB - OBJECTIVE: To compare the prevalence of type 2 diabetes and features of the metabolic syndrome among Filipina and Caucasian women in San Diego County, California. RESEARCH DESIGN AND METHODS: Data on several chronic diseases were collected between 1992 and 1999 from community-dwelling Filipina (n=294) and Caucasian (n=379) women aged 50-69 years. RESULTS: Filipina and Caucasian women did not differ in mean age (59.7 vs. 60 years, respectively), BMI (25.6 vs. 25.4 kg/m(2)), percentage of body fat (33.5 vs. 34.2%), or waist-to-hip ratio (0.84 vs. 0.83), although Filipinas had larger waist circumferences and higher percentages of truncal fat. Compared with Caucasians, Filipinas were less likely to be obese (BMI > or = 30 kg/m(2), 8.8 vs. 14%, P=0.04) and less likely to smoke, consume alcohol, or take postmenopausal estrogen; Filipinas also had lower levels of HDL cholesterol. Compared with Caucasians, Filipinas had higher prevalence of type 2 diabetes by oral glucose tolerance test criteria (36 vs. 9%) and the metabolic syndrome (34 vs. 13%). These differences persisted after adjusting for age, body size, fat distribution, percentage of body fat, smoking, alcohol consumption, exercise, and estrogen therapy. CONCLUSIONS: A total of 10% of Filipinas with diabetes were obese, compared with one third of Caucasians with diabetes. The finding of a high prevalence of diabetes in an unstudied nonobese ethnic group reinforces the importance of expanding the study of diabetes to diverse populations. The high prevalence of diabetes in populations who are not of Northern European ancestry may be missed when they are not obese by Western standards. PMID- 11874937 TI - The effect of estrogen use on levels of glucose and insulin and the risk of type 2 diabetes in american Indian postmenopausal women : the strong heart study. AB - OBJECTIVE: To examine the associations between estrogen use and levels of insulin and glucose as well as the effect of estrogen use on the risk of type 2 diabetes. RESEARCH DESIGN AND METHODS: This report is based on 857 women who were both nondiabetic and postmenopausal at the baseline examination (1989-1992) and who completed a second examination (1993-1995) an average of 4 years later. The participants were divided into three groups: never, past, and current users based on their baseline estrogen use status. ANCOVA was used to compare the insulin and glucose levels among estrogen use groups. Logistic regression was used to evaluate the association between estrogen use and the incidence of type 2 diabetes. RESULTS: Postmenopausal estrogen use was associated with lower fasting glucose (0.2 mmol/l lower) but higher 2-h glucose levels (0.4 mmol/l higher) compared with never users. It was not significantly associated with the risk of type 2 diabetes compared with past and never users, based on American Diabetes Association or World Health Organization definitions of diabetes or on only a 2-h glucose level > or = 11.1 mmol/l. However, the risk of type 2 diabetes increased with increasing duration of estrogen use among current users, with an odds ratio of 1.10 per year of use (95% CI: 1.01-1.19). CONCLUSIONS: The data suggest that estrogen use in American Indian postmenopausal women may relate to deterioration of glucose tolerance. Longer duration of estrogen use among current users may relate to an increased risk of type 2 diabetes. PMID- 11874938 TI - Successful prospective prediction of type 1 diabetes in schoolchildren through multiple defined autoantibodies: an 8-year follow-up of the Washington State Diabetes Prediction Study. AB - OBJECTIVE: Almost 90% of type 1 diabetes appears in individuals without a close family history. We sought to evaluate the best current predictive strategy, multiple defined autoantibodies, in a long-term prospective study in the general population. RESEARCH DESIGN AND METHODS: Autoantibodies to pancreatic islets (islet cell antibodies [ICAs]) and defined autoantibodies (d-aab) to human GAD, IA2/ICA512, and insulin were tested in 4,505 Washington schoolchildren. Eight years later, 3,000 (67%) subjects were recontacted, including 97% of subjects with any test >99th percentile. RESULTS: Six subjects developed diabetes (median interval 2.8 years), all from among the 12 individuals with multiple d-aab, representing 50% positive predictive value (95% CI 25-75%) and 100% sensitivity (58-100%). Among the others, diabetes occurred in 0 of 6 with one d-aab plus ICA, 0 of 26 with ICA only, 0 of 7 with one d-aab equaling the 99th percentile and another d-aab equaling the 97.5th percentile, 0 of 86 with one d-aab, and 0 of 2,863 with no d-aab or ICA. Adjusted for verification bias, multiple d-aab were 99.9% specific (99.86-99.93%). At this age, new d-aab seldom appeared. Once present, d-aab usually persisted regardless of disease progression, although less so for insulin autoantibodies. Insulin secretion by sequential glucose tolerance testing remained normal in four multiple d-aab subjects not developing diabetes. Of children developing diabetes, five of six (83%) would be included if HLA-DQ genotyping preceded antibody testing, but HLA-DQ did not explain outcomes among high-risk subjects, even when considered along with other genetic markers. CONCLUSIONS: Multiple d-aab were established by age 14 years and prospectively identified all schoolchildren who developed type 1 diabetes within 8 years. PMID- 11874939 TI - Diabetes in Ontario: determination of prevalence and incidence using a validated administrative data algorithm. AB - OBJECTIVE: Accurate information about the magnitude and distribution of diabetes can inform policy and support health care evaluation. We linked physician service claims (PSCs) and hospital discharge abstracts (HDAs) to determine diabetes prevalence and incidence. RESEARCH DESIGN AND METHODS: A retrospective cohort was constructed using administrative data from the national HDA database, PSCs for Ontario (population 11 million), and registries carrying demographics and vital statistics. All HDAs and PSCs bearing a diagnosis of diabetes (ICD9-CM 250) were selected for 1991-1999. Two previously reported algorithms for identification of diabetes were applied as follows: "1-claim" (any HDA or PSC showing diabetes) and "2-claim" (one HDA or two PSCs within 2 years showing diabetes). Incident cases were defined as individuals who met the criteria for diabetes for the first time after at least 2 years of observation. For validation, diagnostic data abstracted from primary care charts (n=3,317) of 57 randomly selected physicians were linked to the administrative data cohort, and sensitivity and specificity were calculated. RESULTS: -In 1998, 696,938 individuals met the 1-claim criteria and 528,280 met the 2-claim criteria. Sensitivity for diabetes was 90 and 86%; for the 1- and 2-claim algorithms, specificity was 92 and 97%, respectively, and positive predictive values were 61 and 80%, respectively. Using the 2-claim algorithm, the all-age prevalence increased from 3.2% in 1993 to 4.5% in 1998 (6.1% in adults). Incidence remained stable. CONCLUSIONS: Administrative data can be used to establish population-based incidence and prevalence of diabetes. Diabetes prevalence is increasing in Ontario and is considerably higher than self reported rates. PMID- 11874940 TI - Dose-response effect of pioglitazone on insulin sensitivity and insulin secretion in type 2 diabetes. AB - OBJECTIVE: To investigate the dose-response effects of pioglitazone on glycemic control, insulin sensitivity, and insulin secretion in patients with type 2 diabetes. RESEARCH DESIGN AND METHODS: A total of 58 diet-treated patients with type 2 diabetes (aged 54 +/- 1 years; 34 men and 24 women; BMI 31.5 +/- 0.6 kg/m(2)) were randomly assigned to receive placebo (n=11) or 7.5 mg (n=13), 15 mg (n=12), 30 mg (n=11), or 45 mg (n=11) of pioglitazone per day for 26 weeks. Before and after 26 weeks, subjects underwent a 75-g oral glucose tolerance test (OGTT). RESULTS: Patients treated with 7.5 or 15 mg/day of pioglitazone had no change in fasting plasma glucose (FPG) and fasting plasma insulin (FPI) concentrations or in plasma glucose (PG) and insulin concentrations during the OGTT. Patients treated with 30 and 45 mg/day of pioglitazone, respectively, had significant decreases from placebo in HbA1c (delta=-2.0 and -2.9%), FPG (delta= 66 and -97 mg/dl), and mean PG during OGTT (delta=-84 and -107 mg/dl). Fasting plasma insulin decreased significantly in the 45-mg/day pioglitazone group, but the mean plasma insulin during the OGTT did not change. The insulinogenic index (delta area under the curve [AUC] insulin/deltaAUC glucose) during the OGTT increased significantly in the 30- and 45-mg/day pioglitazone groups (0.13 +/- 0.03 to 0.27 +/- 0.05, P < 0.05). From the OGTT, we previously have derived a composite whole-body insulin sensitivity index (ISI) that correlates well with that measured directly with the insulin clamp technique. Whole-body ISI [ISI=10,000/(square-root (FPG x FPI) x (PG x PI)) where PG and PI equal mean plasma glucose and insulin concentrations during OGTT] increased significantly in patients treated with 30 mg (1.8 +/- 0.3 to 2.5 +/- 0.3, P < 0.05) or 45 mg (1.6 +/- 0.2 to 2.7 +/- 0.6, P < 0.05) per day of pioglitazone. In the basal state, the hepatic ISI [k/(FPG x FPI)[k/(FPG x FPI)], which agrees closely with that measured directly with tritiated glucose, increased in patients treated with 30 mg (0.13 +/- 0.02 to 0.21 +/- 0.03, P < 0.05) and 45 mg (0.11 +/- 0.02 to 0.24 +/ 0.06, P < 0.05) per day of pioglitazone. Significant correlations between the dose of pioglitazone and the changes in HbA1c (r=-0.58), FPG (r=-0.47), mean PG during the OGTT (r=-0.46), insulinogenic index (r=0.34), hepatic ISI (r=0.44), and whole-body ISI (r=0.36) were observed. CONCLUSIONS: Pioglitazone improves glycemic control through the dose-dependent enhancement of beta-cell function and improved whole-body and hepatic insulin sensitivity. PMID- 11874941 TI - alpha-Tocopherol supplementation decreases plasminogen activator inhibitor-1 and P-selectin levels in type 2 diabetic patients. AB - OBJECTIVE: Type 2 diabetic subjects have an increased propensity to premature atherothrombosis. alpha-Tocopherol (AT), a potent antioxidant, has anti inflammatory properties at high doses. The aim of the study was to test the effect of natural (RRR)-AT supplementation (1,200 IU/day) on markers of thrombosis, plasminogen activator inhibitor-1 (PAI-1), and soluble P-selectin (sP selectin) in type 2 diabetic patients with and without macrovascular complications (MVCs) compared with matched control subjects. RESEARCH DESIGN AND METHODS: The volunteers comprised type 2 diabetic patients with (n=23) and without (n=24) MVCs and matched control subjects (n=25). Plasma levels of PAI-1 and P-selectin were assayed at baseline, after 3 months of supplementation, and after a 2-month washout phase. RESULTS: Both diabetic groups had significantly increased levels of PAI-1 compared with control subjects (P < 0.025), whereas only type 2 diabetic patients with MVCs had significantly elevated levels of sP selectin compared with control subjects. AT supplementation significantly lowered levels of PAI-1 and sP-selectin in all three groups. The reduction in PAI-1 levels with AT supplementation was significantly greater in type 2 diabetic patients with MVCs than in those without MVCs (P=0.005). CONCLUSIONS: Thus, AT therapy decreases markers of thrombosis in diabetic patients and control subjects and could be an adjunctive therapy in the prevention of atherosclerosis. PMID- 11874942 TI - Energy metabolism in diabetic and nondiabetic heart transplant recipients. AB - OBJECTIVE: This study examined the metabolic effects of heart transplantation in patients in end-stage cardiac failure. RESEARCH DESIGN AND METHODS: A total of 18 patients after heart transplantation for end-stage heart disease (age 47 +/- 3 years; transplant age 5.5 +/- 1.5 years; BMI 25.8 +/- 0.8 kg/m(2); cyclosporin A 4.2 +/- 0.6 mg/[kg.day]; azathioprine 0.87 +/- 0.31 mg/[kg.day]), 12 patients with type 2 diabetes (D-Tx), and 6 patients without type 2 diabetes (Tx) were studied by means of 1) an oral glucose tolerance test (OGTT) to assess the beta cell secretory response, 2) a euglycemic-hyperinsulinemic (1 mU/[kg.min]) clamp combined with indirect calorimetry and a primed continuous infusion of [6,6 2H2]glucose and [1-13C]leucine to measure postabsorptive and insulin-stimulated carbohydrate and protein metabolism, and 3) 1H-NMR spectroscopy of the calf muscles to measure intramyocellular triglyceride (IMCL) content. The patients were selected from 480 transplant patients in whom there was a 6% prevalence of type 2 diabetes. Five healthy subjects matched for anthropometric parameters served as control subjects (CON). RESULTS: Tx had postabsorptive and insulin stimulated glucose, leucine, and free fatty acid metabolism, as well as IMCL content, similar to that of CON. D-Tx were characterized by a reduced secretory response during the OGTT and peripheral insulin resistance with respect to glucose metabolism, which was paralleled by increased plasma free fatty acid concentrations and IMCL content. A defective insulin-dependent suppression of the endogenous leucine flux (index of proteolysis) was also evident during the clamp in D-Tx. CONCLUSIONS: Heart transplantation, notwithstanding the immunosuppressive therapy, was characterized by a normal postabsorptive and insulin-stimulated glucose, leucine, and free fatty acid metabolism in Tx. In contrast, insulin resistance with respect to glucose, free fatty acids, and protein metabolism was present in D-Tx regardless of whether diabetes was preexisting or consequent to heart transplantation. PMID- 11874943 TI - Plasma F2 isoprostanes: direct evidence of increased free radical damage during acute hyperglycemia in type 2 diabetes. AB - OBJECTIVES: Acute hyperglycemia in type 2 diabetes increases the generation of plasma 8-epi-prostaglandin F2 (8-epi-PGF2alpha) isoprostane, a sensitive direct marker of in vivo free radical oxidative damage to membrane phospholipids. RESEARCH DESIGN AND METHODS: A total of 21 patients with type 2 diabetes underwent an oral 75-g glucose tolerance test. Plasma 8-epi-PGF2alpha isoprostane concentrations (by gas chromatography [GC]/mass spectrometry [MS]), intralymphocyte reduced-to-oxidized glutathione ratios, and plasma total antioxidant capacity were measured at baseline and 90 min after glucose loading. RESULTS: Plasma 8-epi-PGF2alpha isoprostane concentrations rose significantly (P=0. 010) from 0.241 +/- 0.1 to 0.326 +/- 0.17 ng/l after 90 min. Intracellular oxidative balance and plasma antioxidant capacity did not change in either group. CONCLUSIONS: Plasma concentrations of 8-epi-PGF2alpha isoprostane increase during acute hyperglycemia in type 2 diabetes, providing direct evidence of free radical mediated oxidative damage and demonstrating a pathway for an association between acute rather than fasting hyperglycemia and macrovascular risk in type 2 diabetes. PMID- 11874944 TI - Differential effects of metformin and troglitazone on cardiovascular risk factors in patients with type 2 diabetes. AB - OBJECTIVE: Traditional cardiovascular risk factors (CVRF) only partly explain the excessive risk of cardiovascular disease in patients with type 2 diabetes. There is now an increasing appreciation for many novel CVRF that occur largely as a result of insulin resistance and hyperinsulinemia. Therefore, we investigated whether diabetes medications that vary in their mechanism of action and ability to reduce insulin resistance may differ in their effects on both traditional and novel CVRF. RESEARCH DESIGN AND METHODS: We compared the addition of metformin or troglitazone therapy on CVRF in 22 subjects with type 2 diabetes who remained in poor glycemic control (with HbA1c >8.5%) while taking glyburide 10 mg twice daily. Subjects were initially randomized to either metformin 850 mg once daily or troglitazone 200 mg once daily. Both medications were then titrated upward as needed to achieve fasting plasma glucose <120 mg/dl. Measures of glucose control, insulin resistance, and CVRF (blood pressure, lipids, plasminogen activator inhibitor-1, C-reactive protein, fibrinogen, and small dense LDL) were assessed both before and after therapy. RESULTS: After 4 months of treatment, both metformin and troglitazone led to similar decreases in fasting plasma glucose and HbA1c. The reduction in insulin resistance determined by hyperinsulinemic euglycemic clamp was nearly twofold greater with troglitazone than metformin. Metformin did not induce significant changes in blood pressure, LDL cholesterol, LDL size, HDL cholesterol, triglycerides, or plasminogen activator inhibitor-1. However, C-reactive protein did decrease by 33% (6 +/- 1 to 4 +/- 1 mg/l; P < 0.01) [corrected]. Troglitazone therapy was associated with increases in LDL size (26.21 +/- 0.22 to 26.56 +/- 0.25 nm; P=0.04) and HDL cholesterol (33 +/- 3 to 36 +/- 3 mg/dl; P=0.05) and decreases in triglycerides (197 +/- 19 to 155 +/- 23 mg/dl; P=0.07) and C-reactive protein by 60% (8 +/- 3 to 3 +/- 1 mg/l, P < 0.01) [corrected]. CONCLUSIONS: For patients with type 2 diabetes in whom maximal sulfonylurea therapy failed, the addition of the insulin sensitizer troglitazone seemed to have greater benefits on several traditional and novel CVRF than metformin therapy. These differences were not related to glycemic improvement but reflected, in part, the greater reduction in insulin resistance obtained with addition of troglitazone. These data suggest that medications that more effectively address this underlying metabolic defect may be more beneficial in reducing cardiovascular risk in type 2 diabetes. PMID- 11874945 TI - The Gly972Arg polymorphism in insulin receptor substrate-1 is associated with decreased birth weight in a population-based sample of Brazilian newborns. AB - OBJECTIVE: We studied the association between the Gly972Arg polymorphism in insulin receptor substrate-1 (IRS-1) and birth weight in a population-based sample of Brazilian newborns. RESEARCH DESIGN AND METHODS: We studied 194 newborn children with adequate gestational age to identify the association between the Gly972Arg polymorphism and birth weight using PCR-restriction fragment length polymorphism analysis. RESULTS: The data showed that the birth weight was lower in the newborns with the Gly972Arg polymorphism in IRS-1 compared with control subjects (3,141 +/- 31.8 vs. 3,373 +/- 80.3 g, P < 0.008). The results also showed that the frequency of this polymorphism was increased in newborns with a birth weight <3,000 g (P=0.041). CONCLUSIONS: These results suggest that the genotype Gly972Arg may influence birth weight, reinforcing the hypothesis that genetically determined insulin resistance and/or reduced insulin secretion can result in impaired insulin-mediated growth in the fetus. PMID- 11874946 TI - Risk indicators predictive for severe hypoglycemia during the first trimester of type 1 diabetic pregnancy. AB - OBJECTIVES: To investigate the frequency of severe hypoglycemia (SH) and hypoglycemic coma during the first trimester of type 1 diabetic pregnancy and in the 4 months before gestation and to identify risk indicators predicting first trimester SH in a nonselected nationwide cohort of pregnant women with type 1 diabetes. RESEARCH DESIGN AND METHODS: We conducted a longitudinal cohort survey in 278 pregnant type 1 diabetic women using questionnaires at inclusion and at 17 weeks of gestation, addressing the frequencies of SH (i.e., external help required) and hypoglycemic coma, general characteristics, hypoglycemia awareness, blood glucose symptom threshold, and the Hypoglycemia Fear Survey. RESULTS: The occurrence of SH (including hypoglycemic coma) rose from 0.9 +/- 2.4 episodes per 4 months before gestation to 2.6 +/- 6.3 episodes during the first trimester (P < 0.001), including an increase in episodes of coma from 0.3 +/- 1.3 to 0.7 +/- 3.7 (P=0.03). The proportion of women affected by SH rose from 25 to 41% (P < 0.001). First-trimester SH was independently related to a history of SH before gestation (odds ratio [95%CI]: 9.2 [3.9-21.7]), a 10 years' longer diabetes duration (1.6 [1.0-2.4]), an HbA1c level < or = 6.5% (2.5 [1.3-5.0]), and a 0.1 unit/kg higher daily insulin dose (5.4 [1.5-18.9]), adjusted for a decreased symptom threshold. CONCLUSIONS: In type 1 diabetic pregnancy, the risk of SH is increased already before pregnancy and rises further during the first trimester. A history of SH before gestation, longer duration of diabetes, an HbA1c level < or = 6.5%, and a higher total daily insulin dose were risk indicators predictive for SH during the first trimester. Further research should aim to elucidate how the benefits of strict glycemic control balance with the markedly increased risk of SH early in pregnancy. PMID- 11874947 TI - Abnormal carbohydrate metabolism during pregnancy : association with endothelial dysfunction. AB - OBJECTIVE: To evaluate whether abnormal endothelial function, a common finding in premenopausal women with type 2 diabetes, is present in early states of diabetes during pregnancy, such as impaired glucose tolerance (IGT) and gestational diabetes mellitus (GDM). RESEARCH DESIGN AND METHODS: Brachial artery flow mediated dilatation (FMD) (endothelium-dependent) and nitrate-induced dilatation (NID) (endothelium-independent) were measured in 23 pregnant subjects with carbohydrate abnormalities (10 IGT, 13 GDM) and in 15 pregnant control subjects during the third trimester of gestation. High-resolution vascular ultrasonography was used to perform these investigations. A fasting lipid panel was obtained, and glucose and insulin values in response to a 100-g oral glucose load were also measured. RESULTS: FMD was significantly reduced in both groups of women with abnormal carbohydrate metabolism compared with control subjects (7.6 +/- 1.1% in the IGT group and 4.1 +/- 0.9% in the GDM group vs. 10.9 +/- 1.1% in control subjects, P < 0.04 and P < 0.0001, respectively). Significant difference in FMD was also observed between IGT and GDM groups (P < 0.04). NID was comparable in the three groups. Among all subjects, FMD showed a strong independent negative correlation with glycemic area (r=-0.60, P < 0.0001). CONCLUSIONS: Endothelial dysfunction, an early marker of macrovascular disease, is present in pregnancies complicated by IGT and GDM. This alteration, which seems to be directly related to glycemic levels, could explain, at least in part, the increased risk for concurrent hypertensive disorders during pregnancy in these women. PMID- 11874948 TI - Carpal tunnel syndrome in patients with diabetic polyneuropathy. AB - OBJECTIVE: Carpal tunnel syndrome (CTS) and diabetic polyneuropathy (DPN) are common conditions in patients with diabetes and therefore frequently occur concomitantly. Diagnosis of CTS in patients with DPN is important, as therapeutic interventions directed toward relief of CTS may be effective irrespective of diffuse neuropathy. The prevalence of clinical CTS and the most efficient electrodiagnostic discriminators of CTS from diffuse neuropathy are uncertain. RESEARCH DESIGN AND METHODS: A total of 478 subjects, including reference subjects (without diabetes and without neuropathy), nonneuropathic subjects with diabetes, and diabetic subjects with mild, moderate, and severe neuropathy, were evaluated in a cross-sectional design for clinical features of CTS. In the ascertainment of the cohort, a clinical stratification method was used to ensure a broad spectrum of neuropathy severity. All subjects underwent nerve conduction study determinations of median, ulnar, and sural nerve parameters. RESULTS: The prevalence of clinical CTS was 2% in the reference population, 14% in diabetic subjects without DPN, and 30% in those with DPN. Multiple linear regression analysis revealed that mean electrodiagnostic parameters are not significant predictors of clinical CTS in patients with diabetes. Generally, the parameters worsened with severity of neuropathy, but none reliably distinguished diabetic patients with and without CTS. CONCLUSIONS: Given the high prevalence of CTS in patients with DPN and that electrodiagnostic criteria cannot distinguish those with clinical CTS, it is recommended that therapeutic decisions for CTS be made independently of electrodiagnostic findings. PMID- 11874949 TI - Foot ulceration and lower limb amputation in type 2 diabetic patients in dutch primary health care. AB - OBJECTIVE: To determine the incidence of foot ulceration and lower limb amputation in type 2 diabetic patients in primary health care. RESEARCH DESIGN AND METHODS: Data on type 2 diabetes were collected by the Nijmegen Monitoring Project between 1993 and 1998 as part of a study of chronic diseases. The records of all patients recorded as having diabetic foot problems and those who died, moved to a nursing home, or were under specialist care were included. The annual incidence of foot ulceration was defined as the number of type 2 diabetic patients per patient-year who developed a new foot ulcer. Incidence of lower limb amputation was similarly defined. Additional information was collected on treatment of foot ulcers. RESULTS: The study population of type 2 diabetic patients increased from 511 patient-years in 1993 to 665 in 1998. The annual incidence of foot ulceration varied between 1.2 and 3.0% (mean 2.1) per year; 25% of the patients had recurrent episodes. The annual incidence of lower limb amputation varied between 0.5 and 0.8% (mean 0.6). Ten of the 15 amputees died, and 12 of 52 (23%) patients with ulceration had a subsequent amputation or a previous history of amputation. In 35 of the 73 (48%) episodes of ulceration, only the family physician provided treatment. Patients with foot problems were older and had more cardiovascular disease, retinopathy, and absent peripheral pulses. CONCLUSIONS: The incidence of foot ulceration and lower limb amputation in type 2 diabetes is low; nevertheless, recurrence rates of ulceration and risk of amputation are high, with high mortality. PMID- 11874950 TI - Effect of diabetes on renal medullary oxygenation during water diuresis. AB - OBJECTIVES: To study the effect of water diuresis on renal medullary and cortical oxygenation in patients with diabetes using blood oxygenation level--dependent magnetic resonance imaging (BOLD MRI). RESEARCH DESIGN AND METHODS: Nine mild diabetic subjects (48 +/- 2.7 years of age, six women) and nine nondiabetic subjects of similar age and sex, all without known vascular or renal disease, were studied noninvasively by MRI before and during water diuresis. RESULTS: Water diuresis induced an increase in medullary oxygenation in control subjects, producing a decrease in R2* (apparent spin-spin relaxation time) of 1.89 +/- 0.27 (P < 0.01), but no significant change in the group of diabetic subjects. CONCLUSIONS: These findings in middle-aged diabetic subjects, which resembled those previously described in elderly subjects >65 years of age, suggest early impairment of adaptive vasodilatation within the renal medulla in diabetes. PMID- 11874951 TI - Hypertriglyceridemic hyperapoB in type 2 diabetes. AB - OBJECTIVES: Much less attention has been paid to LDL in type 2 diabetes than to VLDL or HDL. In particular, there are few data on apoB levels in these patients. Moreover, most reports have focused on mean lipoprotein levels and consequently there is little information on the frequencies of the various dyslipidemic phenotypes. RESEARCH DESIGN AND METHODS: Plasma and lipoprotein lipids, apoB and apoA1 were measured by standardized methods. LDL particle size was determined by PAGE. The total cohort was divided into phenotypes by two different methods. The first was based on triglycerides (> or = or <1.5 mmol/l) and LDL cholesterol (> or = or <4 mmol/l), whereas the second was based on triglycerides (> or = or <1.5 mmol/l) and apoB (> or = or <120 mg/dl). RESULTS: For the overall cohort, plasma triglycerides were elevated (2.13 +/- 1.6 mmol/l), total and LDL cholesterol were normal (5.34 +/- 1.1 and 3.28 +/- 0.88 mmol/l, respectively), and peak LDL size was reduced (252.9 +/- 5.8 A). HDL cholesterol was between the 25th and 50th percentiles of the general population (1.12 +/- 0.36 mmol/l). The average level of apoB was 114 +/- 29 mg/dl, a value that is between the 50th and 75th percentiles of the general population and is higher than that for LDL cholesterol, which was between the 25th and the 50th percentiles of the population. The results of the phenotyping analysis were as follows. Using the conventional approach, only 23% has abnormal LDL, i.e., an elevated LDL cholesterol level. Using the new approach, almost 40% has an elevated apoB and therefore an elevated LDL particle number. Only 12.8% has combined hyperlipidemia based on the conventional approach, whereas almost one-third had the equivalent, hypertriglyceridemic hyperapoB-based on the new algorithm. The severity of the dyslipoproteinemia in this group was noteworthy. Although the average LDL cholesterol was 3.91 mmol/l, a value just below the 75th percentile of the general population, the average apoB was 145 mg/dl, a value that approximates the 95th percentile of the population. CONCLUSIONS: The dyslipidemic profile of patients with type 2 diabetes is not uniform. A substantial group have normal lipids and normal LDL particle number and size whereas others have markedly abnormal profiles. Diagnosis based on triglycerides and apoB rather than triglycerides and LDL cholesterol revealed that more than one in five had hypertriglyceridemic hyperapoB, which is characterized by hypertriglyceridemia, marked elevation of LDL particle number, small dense LDL, and low HDL, a constellation of abnormalities that is associated with markedly accelerated atherogenesis and therefore justifies intensive medical therapy. PMID- 11874952 TI - Posttransplantation diabetes: a systematic review of the literature. AB - OBJECTIVES: To systematically review the incidence of posttransplantation diabetes (PTD), risk factors for its development, prognostic implications, and optimal management. RESEARCH DESIGN AND METHODS: We searched databases (MEDLINE, EMBASE, the Cochrane Library, and others) from inception to September 2000, reviewed bibliographies in reports retrieved, contacted transplantation experts, and reviewed specialty journals. Two reviewers independently determined report inclusion (original studies, in all languages, of PTD in adults with no history of diabetes before transplantation), assessed study methods, and extracted data using a standardized form. Meta-regression was used to explain between-study differences in incidence. RESULTS: Nineteen studies with 3,611 patients were included. The 12-month cumulative incidence of PTD is lower (<10% in most studies) than it was 3 decades ago. The type of immunosuppression explained 74% of the variability in incidence (P = 0.0004). Risk factors were patient age, nonwhite ethnicity, glucocorticoid treatment for rejection, and immunosuppression with high-dose cyclosporine and tacrolimus. PTD was associated with decreased graft and patient survival in earlier studies; later studies showed improved outcomes. Randomized trials of treatment regimens have not been conducted. CONCLUSIONS: Physicians should consider modification of immunosuppressive regimens to decrease the risk of PTD in high-risk transplant recipients. Randomized trials are needed to evaluate the use of oral glucose-lowering agents in transplant recipients, paying particular attention to interactions with immunosuppressive drugs. PMID- 11874953 TI - Continuous subcutaneous insulin infusion at 25 years: evidence base for the expanding use of insulin pump therapy in type 1 diabetes. AB - Continuous subcutaneous insulin infusion (CSII) is used in selected type 1 diabetic subjects to achieve strict blood glucose control. A quarter of a century after its introduction, world-wide use of CSII is increasing. We review the evidence base that justifies this increase, including effectiveness compared with modern intensified insulin injection regimens and concern about possible complications. Review of controlled trials shows that, in most patients, mean blood glucose concentrations and glycated hemoglobin percentages are either slightly lower or similar on CSII versus multiple insulin injections. However, hypoglycemia is markedly less frequent than during intensive injection therapy. Ketoacidosis occurs at the same rate. Nocturnal glycemic control is improved with insulin pumps, and automatic basal rate changes help to minimize a prebreakfast blood glucose increase (the "dawn phenomenon") often seen with injection therapy. Patients with "brittle" diabetes characterized by recurrent ketoacidosis are often not improved by CSII, although there may be exceptions. We argue that explicit clinical indications for CSII are helpful; we suggest the principal indications for health service or health insurance-funded CSII should include frequent, unpredictable hypoglycemia or a marked dawn phenomenon, which persist after attempts to improve control with intensive insulin injection regimens. In any circumstances, candidates for CSII must be motivated, willing and able to undertake pump therapy, and adequately psychologically stable. Some diabetic patients with well-defined clinical problems are likely to benefit substantially from CSII and should not be denied a trial of the treatment. Their number is relatively small, as would therefore be the demand on funds set aside for this purpose. PMID- 11874954 TI - Behavioral science research in the prevention of diabetes : status and opportunities. AB - Recent studies show diabetes can be prevented. Growing knowledge of its biological bases opens further prevention opportunities. This article focuses on behavioral science research that may advance these opportunities. An ecological model guides attention to how prevention research may be pursued at the individual, group, or community levels. Three key areas are reviewed: risk communication, screening, and preventive interventions. Research on diabetes risk communication is limited but suggests that many are relatively unaware of risks and may have misconceptions about the disease. Amid policy debates and research regarding the potential benefits and costs of screening, identification of diabetes may itself be risky in terms of psychological and social consequences. The Diabetes Prevention Program and other studies make clear that diabetes can be prevented, both by the combination of weight loss and physical activity and by medications. Research needs to address promoting these methods to individuals as well as to groups and even whole communities. Fundamental as well as applied research should address how risks of diabetes are understood and may be communicated; how to enhance benefits and minimize psychological and other risks of screening; how to promote healthy eating and weight loss, physical activity, and appropriate use of medications to prevent diabetes; and how to reduce socioeconomic and cultural disparities in all these areas. PMID- 11874955 TI - Diabetes TV news coverage lags behind. PMID- 11874956 TI - The evidence for the effectiveness of medical nutrition therapy in diabetes management. PMID- 11874957 TI - Treatment issues in type 2 diabetes. PMID- 11874958 TI - Dietary fat and the development of type 2 diabetes. PMID- 11874959 TI - Can we predict type 1 diabetes in the general population? PMID- 11874960 TI - Insulin sensitivity indexes from a single sample in nonobese Japanese type 2 diabetic patients: comparison with minimal model analysis. PMID- 11874961 TI - Status of research funded by the American Diabetes Association: year 3. PMID- 11874962 TI - Fenofibrate lowers plasma triglycerides and increases LDL particle diameter in subjects with type 2 diabetes. PMID- 11874963 TI - Somatostatin therapy in the management of resistant diabetic ketoacidosis. PMID- 11874964 TI - Validation of a case definition for foot complications among hospitalized patients with diabetes. PMID- 11874965 TI - Impairment of the auditory brainstem function in diabetic neuropathy. PMID- 11874966 TI - Plasma homocysteine is not increased in microalbuminuric patients with type 2 diabetes without clinical cardiovascular disease. PMID- 11874967 TI - Continuous subcutaneous insulin infusion to resolve an allergy to human insulin. PMID- 11874968 TI - Lipohypertrophy in young patients with type 1 diabetes. PMID- 11874969 TI - T-cell-mediated autoimmunity may be involved in fulminant type 1 diabetes. PMID- 11874970 TI - Increased risk of diabetes in first-degree relatives of young-onset type 2 diabetic patients compared with relatives of those diagnosed later. PMID- 11874971 TI - Newly diagnosed type 2 diabetic population in Belgium. PMID- 11874972 TI - Response to Jungheim and Koschinsky. PMID- 11874976 TI - Targeting leukemia: from bench to bedside. PMID- 11874977 TI - CpG oligonucleotides: novel regulators of osteoclast differentiation. AB - The macrophage capability to recognize bacterial DNA is mimicked by oligodeoxynucleotides containing unmethylated CG dinucleotides ('CpG' motifs) in specific sequence contexts (CpG ODN). CpG ODN stimulates NF-kappaB activation in murine macrophages. In light of the pivotal role played by NF-kappaB in osteoclast differentiation, we examined the ability of CpG ODN to modulate osteoclastogenesis. CpG ODN alone induced TRAP-positive cells in bone marrow macrophage (BMM) cultures, but not multinucleation or calcitonin receptor expression. CpG ODN inhibited RANKL-induced osteoclastogenesis when present from the beginning of BMM culture, but strongly increased RANKL-induced osteoclastogenesis in RANKL-pretreated BMMs. CpG ODN enhanced the expression of interleukin 1beta (IL-1beta) and tumor necrosis factor alpha (TNF-alpha). Antibodies to TNF-alpha and the TNF type 1 receptor, but not the addition of IL-1 receptor antagonist, blocked CpG ODN-induced osteoclastogenesis in RANKL pretreated cultures. On the other hand, CpG ODN reduced expression of the M-CSF receptor, which is critical during the initiation of osteoclast differentiation. These results suggest that CpG ODN, via the induction of TNF-alpha, support osteoclastogenesis in cells that are committed to the osteoclast differentiation pathway but, due to down-modulation of M-CSF receptor, inhibit early steps of osteoclast differentiation. Thus, CpG ODN represents a potential therapeutic tool for treating bone diseases. PMID- 11874978 TI - A novel doxycycline-inducible system for the transgenic analysis of mammary gland biology. AB - Normal developmental events such as puberty, pregnancy, and parity influence the susceptibility of the mammary gland to tumorigenesis in both humans and rodent model systems. Unfortunately, constitutive transgenic mouse models that rely on mammary-specific promoters to control transgene expression have limited utility for studying the effect of developmental events on breast cancer risk since the hormonal signals governing these events also markedly influence transgene expression levels. A novel transgenic mouse system is described that uses the MMTV-LTR to drive expression of the reverse tetracycline-dependent transactivator rtTA. Transgenic mice expressing rtTA in the mammary epithelium were crossed with reporter lines bearing tet operator-controlled transgenes. We tested the ability to spatially, temporally, and quantitatively control reporter gene expression after administration of doxycycline to bitransgenic mice. Transgene expression using this system can be rapidly induced and deinduced, is highly mammary specific, can be reproducibly titrated over a wide range of expression levels, and is essentially undetectable in the uninduced state. Homogeneous transgene expression throughout the mammary epithelium can be achieved. This system permits transgene expression to be restricted to any desired stage of postnatal mammary gland development. We have developed a mammary-specific, doxycycline-inducible transgenic mouse model for studying the effect of mammary gland development on transgene-mediated phenotypes. Unlike other mammary-specific, transgenic systems that have been described, this system combines spatially homogeneous transgene expression in the mammary epithelium during puberty, pregnancy, lactation, and involution with the use of an orally administered, inexpensive, and widely available inducing agent. This system offers new opportunities for the transgenic analysis of mammary gland biology in vivo. PMID- 11874979 TI - Upstream binding factor up-regulated in hepatocellular carcinoma is related to the survival and cisplatin-sensitivity of cancer cells. AB - Upstream binding factor (UBF) is an RNA polymerase I-specific transcription factor. By representational difference analysis, Northern blot, and cDNA array analysis, up-regulation of UBF was detected in 12 of 17 clinical hepatocellular carcinoma samples comparing to the paired normal liver tissues. Introduction of UBF in human lung fibroblast cells that do not express UBF resulted in an accelerated rate of cell growth; on the other hand, antisense oligodeoxynucleotides (ODNs) treatment of UBF-expressing hepatoma cell lines reduced the level of UBF protein, suppressed the colony formation capacity of these cells on soft agarose, and finally caused cell death. Annexin V binding analysis suggested that anti-UBF ODN-caused cell death might involve weak apoptosis, however, DNA laddering and cleavage of poly (ADP-ribose) polymerase were not observed in these ODN-treated cells. Expression profiling of the anti UBF ODN-treated cells using a human cDNA array revealed that the expression of 30 genes was altered in response to the inhibition of UBF expression. Notably, UBF expression could increase the cell sensitivity to the chemotherapeutic reagent cis-diaminedichloroplatinum (II). We proposed that UBF is fundamental to the survival of cells expressing the gene, and is potential as a target for screening anti-cancer drugs and an indicator in selecting chemotherapeutic reagents. PMID- 11874981 TI - Hsp27 regulates podocyte cytoskeletal changes in an in vitro model of podocyte process retraction. AB - Nephrotic syndrome (NS) is characterized by structural changes in the actin-rich foot processes of glomerular podocytes. We previously identified high concentrations of the small heat shock protein hsp27 within podocytes as well as increased glomerular accumulation and phosphorylation of hsp27 in puromycin aminonucleoside (PAN) -induced experimental NS. Here we analyzed murine podocytes stably transfected with hsp27 sense, antisense, and vector control constructs using a newly developed in vitro PAN model system. Cell morphology and the microfilament structure of untreated sense and antisense transfectants were altered compared with controls. Vector cell survival, polymerized actin content, cell area, and hsp27 content increased after 1.25 microg/ml PAN treatment and decreased after 5.0 microg/ml treatment. In contrast, sense cells were unaffected by 1.25 microg/ml PAN treatment whereas antisense cells showed decreases or no changes in all parameters. Treatment of sense cells with 5.0 microg/ml PAN resulted in increased cell survival and cell area whereas antisense cells underwent significant decreases in all parameters. Hsp27 provided dramatic protection against PAN-induced microfilament disruption in sense > vector > antisense cells. We conclude that hsp27 is able to regulate both the morphological and actin cytoskeletal response of podocytes in an in vitro model of podocyte injury. PMID- 11874980 TI - A novel mechanism for coupling cellular intermediary metabolism to cytosolic Ca2+ signaling via CD38/ADP-ribosyl cyclase, a putative intracellular NAD+ sensor. AB - CD38 is an ectocyclase that converts NAD+ to the Ca2+-releasing second messenger cyclic ADP-ribose (cADPr). Here we report that in addition to CD38 ecto catalysis, intracellularly expressed CD38 may catalyze NAD+-->cADPr conversion to cause cytosolic Ca2+ release. High levels of CD38 were found in the plasma membranes, endoplasmic reticulum, and nuclear membranes of osteoblastic MC3T3-E1 cells. More important, intracellular CD38 was colocalized with target ryanodine receptors. The cyclase also converted a NAD+ surrogate, NGD+, to its fluorescent product, cGDPr (Km approximately 5.13 microM). NAD+ also triggered a cytosolic Ca2+ signal. Similar results were obtained with NIH3T3 cells, which overexpressed a CD38-EGFP fusion protein. The Delta(-49)-CD38-EGFP mutant with a deleted amino terminal tail and transmembrane domain appeared mainly in the mitochondria with an expected loss of its membrane localization, but the NAD+-induced cytosolic Ca2+ signal was preserved. Likewise, Ca2+ release persisted in cells transfected with the Myr-Delta(-49)-CD38-EGFP or Delta(-49)-CD38-EGFP-Fan mutants, both directed to the plasma membrane but in an opposite topology to the full-length CD38-EGFP. Finally, ryanodine inhibited Ca2+ signaling, indicating the downstream activation of ryanodine receptors by cADPr. We conclude that intracellularly expressed CD38 might link cellular NAD+ production to cytosolic Ca2+ signaling. PMID- 11874982 TI - Absence of inducible nitric oxide synthase modulates early reperfusion-induced NF kappaB and AP-1 activation and enhances myocardial damage. AB - The role of nitric oxide (NO) generated by the inducible NO synthase (iNOS) during myocardial ischemia and reperfusion is not understood. We investigated the role of iNOS during early reperfusion damage induced in genetically deficient iNOS (iNOS-/-) mice and wild-type littermates. In wild-type mice, ischemia (60 min) and reperfusion (60 min) induced an elevation in serum levels of creatine phosphokinase and myocardial injury characterized by the presence of scattered apoptotic myocytes and mild neutrophil infiltration. Northern blot analysis showed increased expression of iNOS, whose activity was markedly elevated after reperfusion. Immunohistochemistry showed staining for nitrotyrosine; Western blot analysis showed elevated expression of heat shock protein 70 (HSP70), a putative cardioprotective mediator. Plasma levels of nitrite and nitrate, tumor necrosis factor alpha (TNF-alpha), interleukin 6 (IL-6), and IL-10 were also increased. These events were preceded by degradation of inhibitor kappaBalpha (IkappaBalpha), activation of IkappaB kinase complex (IKK) and c-Jun-NH2-terminal kinase (JNK), and subsequently activation of nuclear factor-kappaB (NF-kappaB) and activator protein 1 (AP-1) as early as 15 min after reperfusion. In contrast, iNOS-/- mice experienced 35% mortality after reperfusion. The extensive myocardial injury was associated with marked apoptosis and infiltration of neutrophils whereas expression of HSP70 was less pronounced. Nitrotyrosine formation and plasma levels of nitrite and nitrate were undetectable. TNF-alpha and IL-6 were increased and IL-10 was reduced in earlier stages of reperfusion. Activation of IKK and JNK and binding activity of NF-kappaB and AP-1 were significantly reduced. Thus, we conclude that iNOS plays a beneficial role in modulating the early defensive inflammatory response against reperfusion injury through regulation of signal transduction. PMID- 11874983 TI - Redistribution of Ca2+ among cytosol and organella during stimulation of bovine chromaffin cells. AB - Recent results indicate that Ca2+ transport by organella contributes to shaping Ca2+ signals and exocytosis in adrenal chromaffin cells. Therefore, accurate measurements of [Ca2+] inside cytoplasmic organella are essential for a comprehensive analysis of the Ca2+ redistribution that follows cell stimulation. Here we have studied changes in Ca2+ inside the endoplasmic reticulum, mitochondria, and nucleus by imaging aequorins targeted to these compartments in cells stimulated by brief depolarizing pulses with high K+ solutions. We find that Ca2+ entry through voltage-gated Ca2+ channels generates subplasmalemmal high [Ca2+]c domains adequate for triggering exocytosis. A smaller increase of [Ca2+]c is produced in the cell core, which is adequate for recruitment of the reserve pool of secretory vesicles to the plasma membrane. Most of the Ca2+ load is taken up by a mitochondrial pool, M1, closer to the plasma membrane; the increase of [Ca2+]M stimulates respiration in these mitochondria, providing local support for the exocytotic process. Relaxation of the [Ca2+]c transient is due to Ca2+ extrusion through the plasma membrane. At this stage, mitochondria release Ca2+ to the cytosol through the Na+/Ca2+ exchanger, thus maintaining [Ca2+]c discretely increased, especially at core regions of the cell, for periods that outlast the duration of the stimulus. PMID- 11874984 TI - Up-regulation of HIV coreceptor CXCR4 expression in human T lymphocytes is mediated in part by a cAMP-responsive element. AB - The chemokine and HIV receptor CXCR4 has been shown to play a role in chemotaxis and HIV-1 entry into T cells. Dibutyryl cAMP (DcAMP), an analog of cAMP, has been shown to increase CXCR4 cell surface expression and HIV-1 infectivity, but the molecular mechanism(s) responsible is unknown. Here we show that DcAMP treatment of purified human T lymphocytes increased transcription of CXCR4 mRNA as well as cell surface and intracellular CXCR4 protein expression. DcAMP-mediated stimulation of human PBL increased T-trophic HIV-1 (X4) fusion and viral replication as measured by syncytia formation and p24 levels, respectively. To determine the region(s) of the CXCR4 promoter required for cAMP responsiveness, truncations and point mutations of the CXCR4 promoter (nucleotides -1098 to +59) fused to luciferase were constructed and transiently transfected into human PBL. Deletional analysis demonstrated that the -1098 to -93 region of the CXCR4 promoter construct could be eliminated; the residual (-93 to +59) promoter retained cAMP responsiveness. Site-directed mutagenesis of a putative cAMP responsive element (CRE) in the 5' UTR (+41 to +49) significantly and specifically attenuated the ability of DcAMP to drive the minimal CXCR4 promoter. Electrophoretic mobility shift assays demonstrated the formation of a complex between the CREB transcription factor and the putative CXCR4 CRE site. Our findings demonstrate a CRE element within the CXCR4 promoter that regulates CXCR4 transcription in response to changes in cAMP signaling. The cAMP-dependent up regulation of CXCR4 mRNA results in increased CXCR4 intracellular and cell surface protein expression as well as increased HIV infectivity. PMID- 11874985 TI - Damage to nuclear DNA induced by Shiga toxin 1 and ricin in human endothelial cells. AB - Ribosome-inactivating proteins (RIPs) remove a specific adenine from 28S rRNA leading to inactivation of ribosomes and arrest of translation. Great interest as to a possible second physiological substrate for RIPs came from the observation that in vitro RIPs remove adenine from DNA. This paper addresses the problem of nuclear lesions induced by RIPs in human endothelial cells susceptible to the bacterial RIP Shiga toxin 1 and the plant RIP ricin. With both toxins, nuclear DNA damage as evaluated by two independent techniques (alkaline-halo assay and alkaline filter elution) appears early, concomitant with (ricin) or after (Shiga toxin 1) the inhibition of protein synthesis. At this time, the annexin V binding assay, caspase 3 activity, the formation of typical < or = 50 Kb DNA fragments, and changes in morphology associated with apoptosis were negative. Furthermore, a block of translation comparable to that induced by RIPs, but obtained with cycloheximide, did not induce nuclear damage. Such damage is consistent with the enzymatic activity (removal of adenine) of RIPs acting in vitro on RNA-free chromatin and DNA. The results unequivocally indicate that RIPs can damage nuclear DNA in whole cells by means that are not secondary to ribosome inactivation or apoptosis. PMID- 11874986 TI - Sodium calcium exchanger plays a key role in alteration of cardiac function in response to pressure overload. AB - The Na+-Ca2+ exchanger (NCX) on the plasma membrane is thought to be the main calcium extrusion system from the cytosol to the extracellular space in many mammalian excitable cells, including cardiac myocytes. However, the pathophysiological role of NCX in the heart is still unclear because of the lack of known specific inhibitors of NCX. To determine the role of NCX in cardiac contraction and the development of cardiac hypertrophy, we imposed pressure overload on the heart of heterozygous NCX knockout (KO) mice by constricting transverse aorta, and examined cardiac function and morphology 3 wk after operation. Although there was no difference in cardiac function between sham operated KO mice and sham-operated wild-type (WT) mice, KO mice showed higher left ventricular pressure and better systolic function than WT mice in response to pressure overload. Northern blot analysis revealed that mRNA levels of sarcoplasmic reticulum Ca2+-ATPase were reduced by pressure overload in left ventricles of WT but not of KO mice. However, hypertrophic changes with interstitial fibrosis were more prominent in KO mice than WT mice. These results suggest that reduction of NCX results in supernormalized cardiac function and causes marked cardiac hypertrophy in response to pressure overload. PMID- 11874987 TI - Cleavage of denatured natural collagen type II by neutrophil gelatinase B reveals enzyme specificity, post-translational modifications in the substrate, and the formation of remnant epitopes in rheumatoid arthritis. AB - During acute inflammation, leukocytes release proteolytic enzymes including matrix metalloproteinases (MMPs), but the physiopathological mechanisms and consequences of this process are not yet fully understood. Neutrophils, the predominant leukocyte type, produce neutrophil collagenase (MMP-8) and gelatinase B (MMP-9) but not the tissue inhibitors of MMPs. After stimulation, these cells also activate MMPs chemically. In arthritic diseases, neutrophils undergo great chemoattraction to the synovium, are activated by interleukin-8, and are stimulated to release gelatinase B in vivo. Production levels and net activities of gelatinase B were found to be absent in degenerative osteoarthritis but significantly increased in rheumatoid arthritis. The cleavage sites in cartilage type II collagen by gelatinase B were determined by a combination of reverse phase high-performance liquid chromatography, Edman degradation, and mass spectrometry analysis. The analysis revealed the site specificity of proline and lysine hydroxylations and O-linked glycosylation, the cleavage specificities by gelatinase B, and the preferential absence and presence of post-translational modifications at P2' and P5', respectively. Furthermore, gelatinase B leaves the immunodominant peptides intact, which are known from studies with (autoreactive) T cells. Lysine hydroxylation was detected at a critical position for T-cell activation. These data lend support to the thesis that extracellular proteolysis and other post-translational modifications of antigenic peptides may be critical in the establishment and perpetuation of autoimmune processes. PMID- 11874988 TI - Disease-associated mutations in KCNE potassium channel subunits (MiRPs) reveal promiscuous disruption of multiple currents and conservation of mechanism. AB - KCNE genes encode single transmembrane-domain subunits, the MinK-related peptides (MiRPs), which assemble with pore-forming alpha subunits to establish the attributes of potassium channels in vivo. To investigate whether MinK, MiRP1, and MiRP2 operate similarly with their known native alpha subunit partners (KCNQ1, HERG, and Kv3.4, respectively) two conserved residues associated with human disease and influential in channel function were evaluated. As MiRPs assemble with a variety of alpha subunits in experimental cells and may do so in vivo, each peptide was also assessed with the other two alpha subunits. Inherited mutation of aspartate to asparagine (D --> N) to yield D76N-MinK is linked to cardiac arrhythmia and deafness; the analogs D82N-MiRP1 and D90N-MiRP2 were studied. Mutation of arginine to histidine (R --> H) to yield R83H-MiRP2 is associated with periodic paralysis; the analogs K69H-MinK and K75H-MiRP1 were also studied. Macroscopic and single-channel currents showed that D --> N mutations suppressed a subset of functions whereas R/K --> H changes altered the activity of MinK, MiRP1, and MiRP2 with all three alpha subunits. The findings indicate that the KCNE peptides interact similarly with different alpha subunits and suggest a hypothesis: that clinical manifestations of inherited KCNE point mutations result from disruption of multiple native currents via promiscuous interactions. PMID- 11874989 TI - Homocystine solubility and vascular disease. AB - There is evidence that mild elevations of tHcy are associated with an increased risk for occlusive vascular disease, thrombosis, and stroke. It is hypothesized here that cellular toxicity could indirectly result from auto-oxidation of homocysteine to homocystine. Elevated levels of total plasma homocysteine could be the primary cause of increased vascular risk, causing endothelial damage through a mechanism similar to that of cystine precipitation, which is known to cause stone formation in cystinosis and cystinuria. In fact, only traces of homocysteine circulate in plasma as the free thiol; the remainder is present as oxidation products. Of these, the symmetric disulfide homocystine is scarcely soluble at neutral pH. Its saturation limit is so close to the concentration of homocysteine in normal plasma that a transient increase of homocysteine levels could lead to precipitation of homocystine microcrystals in the bloodstream. These could damage endothelial tissue, acting as a mechanic primer for subsequent prothrombotic blood vessel alterations. PMID- 11874990 TI - Experimental biology 2002: Translating the genome, April 20-24 New Orleans, Louisiana. PMID- 11874991 TI - Discordant research opportunities and human resources: can we meet the challenge? PMID- 11874992 TI - Artificial neural networks distinguish among subtypes of neoplastic colorectal lesions. AB - BACKGROUND & AIMS: There is a subtle distinction between sporadic colorectal adenomas and cancers (SAC) and inflammatory bowel disease (IBD)-associated dysplasias and cancers. However, this distinction is clinically important because sporadic adenomas are usually managed by polypectomy alone, whereas IBD-related high-grade dysplasias mandate subtotal colectomy. The current study evaluated the ability of artificial neural networks (ANNs) based on complementary DNA (cDNA) microarray data to discriminate between these 2 types of colorectal lesions. METHODS: We hybridized cDNA microarrays, each containing 8064 cDNA clones, to RNAs derived from 39 colorectal neoplastic specimens. Hierarchical clustering was performed, and an ANN was constructed and trained on a set of 5 IBD-related dysplasia or cancer (IBDNs) and 22 SACs. RESULTS: Hierarchical clustering based on all 8064 clones failed to correctly categorize the SACs and IBDNs. However, the ANN correctly diagnosed 12 of 12 blinded samples in a test set (3 IBDNs and 9 SACs). Furthermore, using an iterative process based on the computer programs GeneFinder, Cluster, and MATLAB, we reduced the number of clones used for diagnosis from 8064 to 97. Even with this reduced clone set, the ANN retained its capacity for correct diagnosis. Moreover, cluster analysis performed with these 97 clones now separated the 2 types of lesions. CONCLUSIONS: Our results suggest that ANNs have the potential to discriminate among subtly different clinical entities, such as IBDNs and SACs, as well as to identify gene subsets having the power to make these diagnostic distinctions. PMID- 11874993 TI - Resolution of chronic hepatitis B and anti-HBs seroconversion in humans by adoptive transfer of immunity to hepatitis B core antigen. AB - BACKGROUND & AIMS: Impaired T-cell reactivity is believed to be the dominant cause of chronic hepatitis B virus (HBV) infection. We characterized HBV-specific T-cell responses in chronic hepatitis B surface antigen carriers who received bone marrow from HLA-identical donors with natural immunity to HBV and seroconverted to antibody to hepatitis B surface antigen. METHODS: T-cell reactivity to HBV antigens and peptides was assessed in a proliferation assay, the frequency of HBV core- and surface-specific T cells was quantified directly by ELISPOT assays, and T-cell subsets were analyzed by flow cytometry. RESULTS: CD4+ T-cell reactivity to HBV core was common in bone marrow donors and the corresponding recipients after hepatitis B surface antigen clearance, whereas none reacted to surface, pre-S1, or pre-S2 antigens. Furthermore, CD4+ T cells from donor/recipient pairs recognized similar epitopes on hepatitis B core antigen; using polymerase chain reaction for the Y chromosome, the recipients' CD4+ T lymphocytes were confirmed to be of donor origin. The frequency of core specific CD4+ and CD8+ T cells was several-fold higher than those specific for surface antigen. CONCLUSIONS: This study provides the first evidence in humans that transfer of hepatitis B core antigen-reactive T cells is associated with resolution of chronic HBV infection. Therapeutic immunization with HBV core gene or protein deserves further investigation in patients with chronic hepatitis B. PMID- 11874994 TI - Long-term effect of H2RA therapy on nocturnal gastric acid breakthrough. AB - BACKGROUND & AIMS: Adding histamine 2 receptor antagonists (H2RAs) to proton pump inhibitor (PPI) therapy is a common practice to block nocturnal acid breakthrough (NAB). Controversy exists over its efficacy because of H2RA intolerance. No prospective study has addressed this issue. METHODS: Twenty-three healthy volunteers and 20 gastroesophageal reflux disease (GERD) patients were studied. Ambulatory pH monitoring was performed with one electrode in the gastric fundus and the other 5 cm above the lower esophageal sphincter. Baseline pH testing was performed and repeated after 2 weeks on PPI twice daily before meals (omeprazole 20 mg). All subjects then received 28 days of PPI plus H2RA Qhs (ranitidine 300 mg) with repeat pH testing on days 1, 7, and 28. RESULTS: Eighteen controls and 16 GERD patients completed all 5 studies. Compared with baseline, all 4 medication regimens decreased supine % time pH < 4 (P = 0.001). The administration of PPI + 1 day of H2RA was the only therapy that significantly decreased % time gastric pH < 4 for the supine period compared with PPI twice daily alone (P < 0.001). There was no difference in % time supine gastric pH < 4 between 2 weeks of PPI twice daily alone and either 1 week or 1 month of PPI + bedtime H2RA. CONCLUSIONS: The combination of H2RA and PPI therapy reduced NAB only with the introduction of therapy. Because of H2RA tolerance, there is no difference in acid suppression between PPI twice daily and PPI twice daily + H2RA after 1 week of combination therapy. PMID- 11874995 TI - Surveillance and survival in Barrett's adenocarcinomas: a population-based study. AB - BACKGROUND & AIMS: Guidelines recommend periodic endoscopic surveillance of Barrett's esophagus (BE) patients to detect and treat early esophageal adenocarcinomas; however, no trials or population-based studies exist. We evaluated the association between endoscopic surveillance of BE and survival among esophageal/gastric cardia adenocarcinoma patients. METHODS: We studied a cohort of 23 BE patients, among 589 esophageal or gastric cardia adenocarcinoma patients diagnosed between 1990-1998 at Northern California Kaiser Permanente (a large health maintenance organization). We measured the presence of BE, detection of cancer by endoscopic surveillance, cancer stage, mortality, and potential confounders. RESULTS: BE was diagnosed in 135 of 589 adenocarcinoma patients, with 23 BE patients diagnosed greater than 6 months before cancer was diagnosed. Among these 23 patients, 73% of the surveillance-detected cancer patients (n = 15) were alive at the end of follow-up, compared with none of the patients without surveillance-detected cancers (n = 8; P = 0.001). All surveillance detected cancer patients had low-stage disease and none died directly from cancer. The surveillance/survival association was not substantially altered by stratification for age at BE diagnosis or other potential confounders. CONCLUSIONS: Surveillance-detected BE-associated adenocarcinomas were associated with low-stage disease and improved survival. Additional studies are needed to evaluate potential biases and whether screening/surveillance programs decrease mortality among all patients in surveillance. Few patients (3.9%) had a BE diagnosed before their cancer. Thus, even if current surveillance techniques are effective, they are unlikely to substantially impact the population's mortality from esophageal cancer; better methods are needed to identify at risk patients. PMID- 11874996 TI - Long-term treatment with sulindac in familial adenomatous polyposis: a prospective cohort study. AB - BACKGROUND & AIMS: Management of patients with familial adenomatous polyposis (FAP) can consist of colectomy with ileorectal anastomosis (IRA). Sulindac, a nonsteroidal anti-inflammatory drug, causes regression of colorectal adenomas in the retained rectal segment of FAP patients, although long-term use of this therapy has not been studied. We evaluated the long-term effectiveness and toxicity of sulindac in attempting to maintain retained rectal segments free of adenomas. METHODS: Twelve FAP patients (5 women), mean age 37.1 years, with IRA received sulindac (mean dosage, 158 mg/day) for a mean period of 63.4 +/- 31.3 months (range, 14-98 months). Number, size, and histologic grade of polyps, side effects, and medication compliance were assessed every 4 months. RESULTS: Seven of 12 patients (58%) remained in the study (6 of these polyp-free) for a mean of 76.9 +/- 27.5 months. Five of 12 patients (42%) withdrew from the trial after a mean follow-up period of 44 +/- 28 months (range, 14-89 months). A significant regression of polyp number was observed in all patients at 12 months (P = 0.039) and at a mean of 63.4 +/- 31.3 months (P = 0.006). Prevention of recurrence of higher-grade adenomas (tubulovillous, villous adenomas) was also observed (P = 0.004). At 35 months of follow-up, 1 patient developed stage III cancer in the rectal stump. The most common side effect was rectal mucosal erosions in 6 patients. CONCLUSIONS: Long-term use of sulindac seems to be effective in reducing polyp number and preventing recurrence of higher-grade adenomas in the retained rectal segment of most FAP patients. Erosions at the IRA site can preclude adequate dose maintenance. PMID- 11874997 TI - A population-based study of the biochemical and clinical expression of the H63D hemochromatosis mutation. AB - BACKGROUND & AIMS: Two major mutations are defined within the hemochromatosis gene, HFE. Although the effects of the C282Y mutation have been well characterized, the effects of the H63D mutation remain unclear. We accessed a well-defined population in Busselton, Australia, and determined the frequency of the H63D mutation and its influence on total body iron stores. METHODS: Serum transferrin saturation and ferritin levels were correlated with the H63D mutation in 2531 unrelated white subjects who did not possess the C282Y mutation. RESULTS: Sixty-two subjects (2.1%) were homozygous for the H63D mutation, 711 (23.6%) were heterozygous, and 1758 (58.4%) were wild-type for the H63D mutation. Serum transferrin saturation was significantly increased in male and female H63D homozygotes and heterozygotes compared with wild-types. Serum ferritin levels within each gender were not influenced by H63D genotypes. Elevated transferrin saturation > or = 45% was observed in a greater proportion of male H63D carriers than male wild-types. Male H63D homozygotes (9%) and heterozygotes (3%) were more likely to have both elevated transferrin saturation and elevated ferritin > or = 300 ng/mL than male wild-types (0.7%). Homozygosity for H63D was not associated with the development of clinically significant iron overload. CONCLUSIONS: Presence of the H63D mutation results in a significant increase in serum transferrin saturation but does not result in significant iron overload. In the absence of the C282Y mutation, the H63D mutation is not clinically significant. PMID- 11874998 TI - Primary biliary cirrhosis: incidence and predictive factors of cirrhosis development in ursodiol-treated patients. AB - BACKGROUND & AIMS: Ursodeoxycholic acid (UDCA) slows the progression of primary biliary cirrhosis (PBC). However, some UDCA-treated patients escape and progress toward cirrhosis and end-stage disease. This study aimed to assess the incidence of cirrhosis in UDCA-treated patients with PBC and to determine the predictive factors of cirrhosis development under this treatment. METHODS: A Markov model was used to describe the progression toward cirrhosis in 183 UDCA-treated patients with PBC. A total of 254 pairs of liver biopsy specimens collected during 655 patient-years were studied. RESULTS: The incidence of cirrhosis after 5 years of UDCA treatment was 4%, 12%, and 59% among patients followed-up from stages I, II, and III, respectively. At 10 years, the incidence was 17%, 27%, and 76%, respectively. The median time for developing cirrhosis from stages I, II, and III was 25 years, 20 years, and 4 years, respectively. The independent predictive factors of cirrhosis development were serum bilirubin greater than 17 mumol/L, serum albumin less than 38 g/L, and moderate to severe lymphocytic piecemeal necrosis. CONCLUSIONS: This study provides new data about the time course of PBC under UDCA and constitutes a rationale for the design and evaluation of clinical trials aimed to assess the efficacy of drugs associated with UDCA. PMID- 11874999 TI - TRAIL and its receptors in the colonic epithelium: a putative role in the defense of viral infections. AB - BACKGROUND & AIMS: Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a member of the tumor necrosis factor family and induces apoptosis by cross-linking either of the 2 TRAIL receptors containing a death domain (TRAIL-R1 or TRAIL-R2). TRAIL-R3 and TRAIL-R4 are receptors that do not transmit an apoptotic signal. The aim of this study was to investigate the expression and function of TRAIL and its receptors in normal colonic epithelium. METHODS: TRAIL and TRAIL receptor expression was studied by reverse-transcriptase polymerase chain reaction and immunohistochemistry. TRAIL sensitivity of epithelial cells was determined in vitro. RESULTS: Normal colonic epithelial cells express TRAIL, TRAIL-R1, TRAIL-R2, and TRAIL-R4. Interestingly, TRAIL and TRAIL-R2 are coexpressed mostly in the luminal surface epithelium. Despite the expression of apoptosis-mediating TRAIL receptors, the normal colonic crypt epithelium is completely resistant to TRAIL-induced apoptosis in vitro. Using an infection model with restricted human cytomegalovirus gene expression or productive adenovirus infection in the colon carcinoma cell line Caco-2, we show that TRAIL sensitivity of colonic epithelial cells is induced on virus infection along with an up-regulation of TRAIL-R1 and TRAIL-R2 on the cell surface. CONCLUSIONS: We conclude that the TRAIL system may play a role in the early elimination of virus infected epithelial cells in the normal gut. PMID- 11875000 TI - Ubiquitous production of macrophage migration inhibitory factor by human gastric and intestinal epithelium. AB - BACKGROUND & AIMS: Macrophage migration inhibitory factor (MIF) inhibits macrophage migration and has pleiotropic activities on immune and inflammatory responses, cell growth, and glucose metabolism. MIF is produced by T cells, macrophages, and endothelial cells. Because intestinal epithelial cells produce mediators important for regulating mucosal immune and inflammatory responses, we sought to determine if these cells produce MIF. METHODS: MIF expression was determined by immunostaining of human intestinal mucosa, intestinal xenografts, and cultured cells. MIF protein levels were quantitated by enzyme-linked immunosorbent assay and immunoblot analysis, messenger RNA was assessed by real time reverse-transcription polymerase chain reaction, and functional activity was assessed by enzymatic and migration assays. RESULTS: MIF was abundantly expressed in vivo in gastric, small intestinal, and colonic epithelium and in epithelium lining human intestinal xenografts. MIF was also constitutively expressed at the messenger RNA and protein level by several cultured colon and gastric epithelial cell lines, and its expression in those cells was not up-regulated by the proinflammatory cytokines interleukin 1alpha, tumor necrosis factor alpha, or interferon gamma. Epithelial MIF from cultured cells was released predominantly from the apical side after Salmonella infection, had tautomerase activity, and arrested macrophage migration. CONCLUSIONS: Human intestinal epithelial cells are a major source of MIF, a molecule that can regulate macrophage migration, inflammation, and cell metabolism. PMID- 11875001 TI - Increased sensitivity to the locomotor-activating effects of corticotropin releasing hormone in cholestatic rats. AB - BACKGROUND & AIMS: Fatigue is a common complaint of patients with cholestatic liver disease. Defective central corticotropin-releasing hormone (CRH) release has been postulated as playing a role in the genesis of fatigue and decreased hypothalamic CRH expression has been identified in an animal model of cholestatic liver injury. Therefore, we hypothesized that reduced central CRH release contributes to fatigue associated with cholestatic liver disease and tested this hypothesis in cholestatic rats. METHODS: Locomotor activity during prolonged observation, measured by using an infrared beam activity monitor, was used as a surrogate marker of fatigue or fatigability. Rats with cholestasis secondary to bile duct resection (BDR) had significantly lower basal locomotor activity compared with sham controls. RESULTS: Intracerebroventricular injections of CRH (0.05, 0.1, 1.0 microg/rat) caused significantly greater locomotor activation in BDR animals than controls. In BDR rats, this locomotor activation was blocked by the coadministration of the nonspecific CRH-receptor antagonist astressin (25 microg/rat) and the specific CRH type 1-receptor antagonist NBI-27941 (10 microg/rat). Immunoblotting showed a dramatic increase in hypothalamic CRH type 1 receptor expression in BDR rats compared with controls, which was paralleled by a striking reduction in hypothalamic CRH levels. CONCLUSIONS: These findings are consistent with defective central CRH neurotransmission contributing to decreased locomotor activity in cholestatic rats and have direct implications for cholestasis-associated fatigue. PMID- 11875002 TI - Cdx2 ectopic expression induces gastric intestinal metaplasia in transgenic mice. AB - BACKGROUND & AIMS: Intestinal-type gastric cancer is often preceded by intestinal metaplasia in humans. The genetic events responsible for the transdifferentiation that occurs in intestinal metaplasia are not well understood. Cdx2, a transcription factor whose expression is normally limited to the intestine, has been detected in gastric intestinal metaplasia. Cdx2 induces differentiation of intestinal epithelial cells in vitro; therefore, we sought to establish whether a causal relationship exists between Cdx2 activation and intestinal metaplasia. METHODS: Cdx2 expression was directed to the gastric mucosa in transgenic mice using cis-regulatory elements of Foxa3 (Hnf3gamma). Transgenic mice were analyzed for histologic and gene expression changes. RESULTS: Histologic examination of the gastric mucosa of the Foxa3/Cdx2 mice revealed the presence of alcian blue positive intestinal-type goblet cells, a hallmark of intestinal metaplasia. In addition, Cdx2 induced the expression of intestine-specific genes. CONCLUSIONS: Gastric expression of Cdx2 alone was sufficient to induce intestinal metaplasia in mice. These mice represent a powerful tool to investigate the molecular mechanisms that promote intestinal metaplasia. Moreover, as gastric cancer in humans is often preceded by intestinal metaplasia, the phenotype described here strongly suggests involvement of Cdx2 in the initiation of the process leading to intestinal neoplasia of the gastric mucosa. PMID- 11875003 TI - Induction of anaphylatoxin C5a receptors in rat hepatocytes by lipopolysaccharide in vivo: mediation by interleukin-6 from Kupffer cells. AB - BACKGROUND & AIMS: In normal rat liver, anaphylatoxin C5a induces glucose output from hepatocytes indirectly via prostanoids released from Kupffer cells. Correspondingly, it was found that hepatocytes, in contrast to Kupffer cells, did not express C5a receptors. Lipopolysaccharide (LPS) has been reported to enhance C5a receptor expression in murine livers. This might be the result of de novo expression in hepatocytes. METHODS: C5a receptor expression was investigated in hepatocytes after in vivo treatment of rats with LPS and in vitro stimulation of isolated cells with LPS and proinflammatory cytokines on messenger RNA (mRNA) and protein level, and functionally in isolated hepatocytes and perfused liver. RESULTS: In vivo treatment of rats with LPS induced C5a receptor mRNA and protein in hepatocytes with a maximum after 8-10 hours. At this time-point, C5a directly activated glycogen phosphorylase in isolated hepatocytes and enhanced glucose output in perfused livers without the involvement of prostanoids. LPS failed to induce C5a receptors in cultured hepatocytes in vitro, whereas interleukin (IL) 6 and IL-1beta, which are known to be released from Kupffer cells on stimulation with LPS, did so. In cocultures of hepatocytes with Kupffer cells, LPS induced C5a receptors in hepatocytes in an IL-6-dependent manner. CONCLUSIONS: Thus, IL-6 from Kupffer cells appears to be the main mediator of LPS-induced de novo expression of C5a receptors in hepatocytes. PMID- 11875004 TI - Down-regulated in adenoma mediates apical Cl-/HCO3- exchange in rabbit, rat, and human duodenum. AB - BACKGROUND & AIMS: Duodenal bicarbonate secretion is in part mediated by an apical Cl-/HCO3- exchanger of unknown molecular nature. The recently discovered dra (down-regulated in adenoma) gene encodes a transport protein (DRA) for SO4(2 ), Cl-, and HCO3-. The aim of this study was to investigate whether DRA may be the duodenal apical Cl-/HCO3- exchanger. METHODS: DRA, Na+/H+ exchanger (NHE) isoform 3, and anion exchanger isoform (AE) 2 messenger RNA expression levels were studied in rat, rabbit, and human gastrointestinal tract by semiquantitative reverse-transcription polymerase chain reaction and in situ hybridization (DRA in human intestine). The subcellular localization of DRA was determined by Western analysis and immunohistochemistry. Using rabbit and rat duodenal brush border membrane vesicles, anion exchange characteristics were investigated. RESULTS: DRA expression was high in duodenum and colon of all species, whereas NHE3 messenger RNA expression was low in duodenum and high in colon. Western analysis and immunohistochemistry showed an apical localization for DRA. Rabbit and rat duodenal brush border membrane vesicles showed 4,4'-diisothiocyanostilbene-2,2' disulfonic acid-sensitive Cl-/Cl-, HCO3-/Cl-, SO4(2-)/Cl-, and Cl-/SO4(2-) exchange, with evidence for one major brush border membrane Cl-/anion exchanger, an affinity for Cl- > HCO3-, and a much higher affinity for SO4(2-) in rat than rabbit. The strong predominance of DRA over NHE3 and NHE2 expression in duodenum was paralleled by much higher Cl-/HCO3- than Na+/H+ exchange rates in brush border membrane vesicles and likely explains the high duodenal HCO3- secretory rates. CONCLUSIONS: These data suggest that DRA is the major apical anion exchanger in the duodenum as well as the colon and the likely transport protein for duodenal electroneutral HCO3- secretion. PMID- 11875005 TI - Genetic deficiency in the chemokine receptor CCR1 protects against acute Clostridium difficile toxin A enteritis in mice. AB - BACKGROUND & AIMS: The role of the CC chemokine receptor (CCR) 1 in acute enteritis was investigated by subjecting CCR1 knockout mice to Clostridium difficile toxin A treatment. METHODS: Toxin A or vehicle was injected into ileal loops in anesthetized wild-type, CCR1-/- and macrophage inhibitory protein (MIP) 1alpha-/- mice. After 1-4 hours, fluid accumulation was calculated, and the loops were processed for histology, myeloperoxidase activity, regulated on activation, normal T cell expressed and secreted (RANTES) production, and messenger RNA measurements. RESULTS: Toxin A induced in all mice a significant (P < 0.05) increase in ileal fluid accumulation, epithelial damage, and neutrophil infiltration, with all parameters being significantly (P < 0.01) lower in CCR1-/- and MIP-1alpha-/- mice. Ileal messenger RNA expression of the CCR1 ligands MIP 1alpha and RANTES and RANTES synthesis were increased in toxin A-treated wild type mice. The RANTES antagonist Met-RANTES significantly (P < 0.01) reduced the toxin A-induced increases in ileal fluid accumulation and myeloperoxidase activity in wild-type mice. CONCLUSIONS: C. difficile toxin A-induced murine enteritis involves CCR1 and its ligands MIP-1alpha and RANTES, which may be important mediators of the neutrophil recruitment characterizing acute, enterotoxin-mediated enteritis. PMID- 11875006 TI - Intravital observation of adhesion of lamina propria lymphocytes to microvessels of small intestine in mice. AB - BACKGROUND & AIMS: Although the recirculation of lymphocytes through the intestinal mucosa is important for the specific immune defense, the homing of lamina propria lymphocytes (LPLs) has not been clearly understood. The aim of this study is to compare, under an intravital microscope, the dynamic process of lymphocyte-endothelium recognition and binding in the murine intestinal mucosa of T lymphocytes from the lamina propria of intestine to that of T lymphocytes from the spleen. METHODS: LPLs isolated from nonlymphoid areas of the small intestine and spleen (SPL) were fluorescence-labeled and injected into a jugular vein of recipient mice. Microvessels of the villus mucosa and ileal Peyer's patches were observed under an intravital fluorescence microscope, and the effects of anti adhesion-molecule antibodies on lymphocyte-endothelial interaction were investigated. RESULTS: LPLs accumulated abundantly in the microvessels of villus tips but not in the submucosal venules or postcapillary venules of Peyer's patches, where SPLs migrated selectively. The accumulation of LPLs in the villus tips was almost completely inhibited by anti-beta7-integrin and was significantly inhibited by anti-mucosal addressin cell-adhesion molecule 1 (MAdCAM-1) and anti alpha4-integrin. Significant MAdCAM-1 expression was observed in the microvessels of the villus mucosa. Some SPLs adhered to the nonlymphoid mucosa, but most soon detached. CONCLUSIONS: It was shown in vivo for the first time that T lymphocytes from the lamina propria but not from the spleen adhere selectively, mostly via alpha4beta7 and MAdCAM-1, to the microvessels of villus tips of the intestine, but not to the postcapillary venules of Peyer's patches. PMID- 11875007 TI - Inactivation of the hemochromatosis gene differentially regulates duodenal expression of iron-related mRNAs between mouse strains. AB - BACKGROUND & AIMS: Hfe knockout mice, like patients with hereditary hemochromatosis, have augmented duodenal iron absorption and increased iron deposition in hepatic parenchymal cells. The goals of the present study were to gain further insight into the control of iron absorption by comparing the transcript levels of iron-related genes in the duodenum of DBA/2 Hfe-/- mice, susceptible to iron loading, and wild-type controls, and to test whether variations in the duodenal expression of these messengers contribute to the DBA/2 and C57BL/6 strain differences in the severity of hepatic iron loading. METHODS: Expression of the different transcripts was quantified by real-time polymerase chain reaction. RESULTS: The 2 strains differ strikingly, not only in the severity of hepatic iron loading, but also in the duodenal expression of iron related genes. In DBA/2 Hfe-/- mice, increased intestinal iron absorption results from the concomitant up-regulation of the Dcytb, DMT1, and FPN1 messengers. No increase in the expression of these messengers is seen in C57BL/6 Hfe-/- mice. CONCLUSIONS: The up-regulation of these transcripts suggests that an inappropriate iron-deficiency signal is sensed by the duodenal enterocytes, leading to an enhanced ferric reductase activity and the increase of duodenal iron uptake and transfer to the circulation. The genes modifying the hemochromatosis phenotype probably act by modifying the expression of these 3 messengers. PMID- 11875008 TI - Mild gastritis alters voltage-sensitive sodium currents in gastric sensory neurons in rats. AB - BACKGROUND & AIMS: Visceral hypersensitivity can be found in more than one third of patients with dyspeptic symptoms. We hypothesized that peripheral sensitization plays an important role in the development of hypersensitivity. METHODS: We induced mild gastritis in Sprague-Dawley rats by adding 0.1% iodoacetamide to the drinking water. The stomach was injected with a retrograde label to identify gastric sensory neurons. Nodose and T9, T10 dorsal root ganglia were removed 7 days after initiation of iodoacetamide treatment. The cells were dissociated and cultured for 3-8 hours before recording whole cell currents using the patch-clamp technique. RESULTS: Iodoacetamide induced a mild gastritis. Although there were no changes in voltage-sensitive inward and outward currents in nodose neurons, the inward currents increased significantly in T9, T10 spinal neurons. A more detailed analysis of sodium currents showed that this was caused by an increase in the tetrodotoxin-resistant sodium current. CONCLUSIONS: Mild gastritis increases the tetrodotoxin-resistant current in gastric spinal sensory neurons. Considering the importance of sodium currents as determinants of neuron excitability, this change may contribute to peripheral sensitization and enhanced neuron excitability. PMID- 11875009 TI - CD40/CD154 ligation is required for the development of acute ileitis following oral infection with an intracellular pathogen in mice. AB - BACKGROUND & AIMS: Acute inflammatory ileitis occurs in C57BL/6 mice after oral infection with Toxoplasma gondii. We evaluated the role of CD40/CD154 interaction in the development of acute ileitis in this experimental model. METHODS: CD154-/- and anti-CD154 antibody-treated mice as well as chimeric mice, either C57BL/6 or CD40-/- reconstituted with bone marrow from C57BL/6 or CD40-/- mice, were orally infected with cysts. Inflammation was assessed by qualitative histologic and phenotypic analysis of the intestinal compartment at day 7 after infection. Intestinal chemokine and cytokine production was assayed by ribonuclease protection assay. RESULTS: CD154-/- and anti-CD154 monoclonal antibody-treated mice failed to develop an acute, lethal ileitis after oral infection and survived. Chimeric mice reconstituted with bone marrow from C57BL/6 mice developed ileitis and died, whereas those recipient mice deficient in CD40 survived. CD40 expression in the intestine after infection was found principally within the B-cell compartment. A modest increase in CD40 expression in both the macrophage and dendritic cell compartments was also observed. Both chemokine and cytokine expression was up-regulated in those recipients of wild-type bone marrow. Impairment of CD40/CD154 interaction abrogated the production of these proinflammatory productions. CONCLUSIONS: CD40/CD154 interaction is essential to the development of inflammation in this pathogen-driven experimental model of acute ileitis. PMID- 11875010 TI - Multiple mechanisms in indomethacin-induced impairment of hepatic cytochrome P450 enzymes in rats. AB - BACKGROUND & AIMS: Indomethacin impairs liver microsomal monooxygenase activities mediated by cytochrome P450 (CYP). We investigated the inhibition mechanism and the isoform selectivity in vitro and in vivo. METHODS: In an in vitro study, liver microsomes from male Wistar rats were preincubated with indomethacin and a reduced nicotinamide adenine dinucleotide phosphate-generating system, followed by assay of monooxygenase activities indicative of several CYP isoforms. In an in vivo study, rats were intraperitoneally treated with indomethacin, followed by preparation of microsomes and the enzyme assays. RESULTS: The preincubation of microsomes with indomethacin and reduced nicotinamide adenine dinucleotide phosphate decreased CYP3A2 activity but not any other isoforms. Kinetic analysis showed the mechanism-based inactivation of CYP3A2. The metabolism of [14C]indomethacin resulted in covalent binding to microsomal protein, which was diminished by inhibiting CYP3A enzyme. Administration of indomethacin caused impairment of not only CYP3A2 but also other CYP isoforms. Rats were protected from the impairment of the CYP enzymes except CYP3A2 by depleting macrophages and inhibiting inducible nitric oxide synthase. CONCLUSIONS: Metabolism of indomethacin causes inactivation of CYP3A2, which is the result of the covalent binding of its metabolite, whereas partially selective in vivo impairment of CYP isoforms is suggested to be indirect inhibition by inflammatory mediators probably released from Kupffer cells. PMID- 11875011 TI - Molecular evidence for the same clonal origin of both components of an adenosquamous Barrett carcinoma. AB - We describe an uncommon case of adenosquamous carcinoma arising in a Barrett esophagus in a 72-year-old white man who occasionally used alcohol, and was a nonsmoker for 34 years. Polymerase chain reaction-based microsatellite analysis was performed on the adenocarcinoma component (AC) and squamous cell carcinoma component (SC) of the tumor. The metaplastic Barrett epithelium (BE), the AC and the SC all showed loss of the same allele at 4 markers on chromosome 9p. Furthermore, the AC and the SC both showed loss of the same allele at all informative markers tested on chromosomal arms 3p, 5q, 10q, 14q, and 18q. In addition, both the SC and AC component contained the same missense mutation in the p53 tumor-suppressor gene. The only observed difference was a shift at a marker on chromosome 16q in the AC, whereas no shift was found in the BE and the SC. These findings suggest that this biphasic tumor has a monoclonal origin. The divergence presumably occurred late in the tumorigenesis of this carcinoma. PMID- 11875012 TI - A homozygous HFE gene splice site mutation (IVS5+1 G/A) in a hereditary hemochromatosis patient of Vietnamese origin. AB - The vast majority of Caucasian patients presenting with hereditary hemochromatosis demonstrate a single homozygous missense mutation in the HFE gene (C282Y). The underlying genetic defects in hemochromatosis patients of non Caucasian origin are largely unknown. A 48-year-old man of Vietnamese origin presented with insulin-dependent diabetes mellitus, tertiary adrenocortical insufficiency, and laboratory results highly indicative of hereditary hemochromatosis. Because the patient was negative for the known HFE gene mutations C282Y, H63D, and S65C HFE, the entire coding region and intron/exon boundaries of the HFE gene was investigated. Sequencing studies identified a homozygous G-to-A transition at position +1 of intron 5 (IVS5+1 G/A). This newly described mutation alters the invariant G at position +1 of the 5' splice site causing altered mRNA splicing and exon skipping with exon 4 being spliced to exon 6. Both heterozygously affected children (age 19 and 20 years) had moderately increased ferritin levels with normal serum iron concentration and transferrin saturation. The newly described mutation was not detected in a control group consisting of 220 Caucasian individuals as verified by allele-specific polymerase chain reaction. We describe for the first time a homozygous HFE splice site mutation (IVS5+1 G/A) in a non-Caucasian patient with hereditary hemochromatosis. Although the absence of this novel HFE gene mutation in Caucasian subjects suggests that the mutation is exclusive to this family, mutation screening in populations of different ethnic background is recommended to precisely define its contribution to hereditary hemochromatosis in non-Caucasian patients. PMID- 11875013 TI - A syndrome of hereditary pancreatic adenocarcinoma and cysts of the liver and kidneys. AB - Pancreatic adenocarcinoma is the fifth leading cause of cancer death in developed countries. Several family and population studies have suggested that there is a genetic predisposition in about 10% of cases. Despite this, pedigrees showing a definite Mendelian inheritance pattern are quite rare. Recently, a family came to our attention with several cases of pancreatic adenocarcinoma. A detailed family medical history was obtained from the index patient. Medical records, including death certificates, histopathology, radiology, and laboratory reports from several family members were reviewed. Computerized tomography scans and CA19-9 serum assays were performed on selected family members. The family seems to have a syndrome of autosomal dominant adenocarcinoma of the pancreas, accompanied by multiple cysts of the liver and kidneys. Affected family members without pancreatic cancer have elevated serum CA19-9 levels. This seems to be a previously undescribed syndrome. The family may be carrying a tumor suppressor gene mutation specific for pancreatic adenocarcinoma. PMID- 11875014 TI - Successful infliximab treatment for steroid-refractory celiac disease: a case report. AB - Celiac disease is a T cell-mediated enteropathy induced by gluten in genetically predisposed individuals. The majority of patients responds to a gluten-free diet but a small number do not. After the exclusion of gluten in the diet, ulcerative jejunititis, and an enteropathy-associated T-cell lymphoma, another treatment modalities, such as systemic steroids and immunosuppressives, may be necessary. This article reports the case of a 47-year-old white woman with immunoglobulin A deficiency. She was diagnosed with celiac disease with subtotal villous atrophy on jejunal biopsy together with positive antiendomysium and antigliadin immunoglobulin G antibodies. Despite close adherence to a gluten-free diet, her weight continued to decrease, she had diarrhea, and her distal duodenal histology showed no improvement. Some improvement in her symptoms was observed with cyclosporine and systemic steroids, but this was not sustained. Recent evidence has suggested that anti-tumor necrosis factor alpha antibodies have a role in the amelioration of an animal model of villous atrophy, and after careful consideration, she was treated with infliximab. There was a dramatic improvement in her weight, symptoms, and distal duodenal histology. The response has been maintained for 18 months while on azathioprine therapy. It is concluded that infliximab is an effective treatment that may be considered in a small number of patients with refractory celiac disease, resistant to other therapy. PMID- 11875015 TI - Tissue plasminogen activator is required for the growth, invasion, and angiogenesis of pancreatic tumor cells. AB - BACKGROUND & AIMS: Overexpression of tissue-type plasminogen activator (t-PA) in exocrine pancreatic tumors might be a determinant of the aggressive biological behavior of these tumors. METHODS: Endogenous t-PA production was suppressed by antisense oligonucleotides or transcripts in CAPAN-1 and RWP-1 cell lines. Reciprocally, the t-PA non-expressing BxPC-3 and PANC-1 cells were stably transfected to overexpress t-PA. Recombinant t-PA and chemical inhibitors were also used on these cells. Clones were assayed for invasion and growth in vitro and in vivo. RESULTS: In vitro, specific inhibition of t-PA expression or activity significantly inhibited the proliferation of t-PA-producing RWP-1, CAPAN 1, and SK-PC-1 cells. Antisense constructs were used to generate RWP-1 clones stably suppressed for t-PA expression (AS clones). These clones had a significantly reduced invasion and proliferation on plastic and in soft agar. The addition of recombinant t-PA rescued the growth of the AS clones to parental levels and was mitogenic for other independent pancreas cell lines. This effect did not require plasmin activity. In athymic mice, RWP-1 AS clones produced tumors fivefold smaller than control clones. AS tumors contained a significantly reduced number of Ki67-positive nuclei, fewer mitotic cells, and a remarkably reduced angiogenic network. Finally, the generation of tetracycline-repressed t PA transfectants in PANC-1 cells confirmed the activity of t-PA in invasion and proliferation in vitro and in vivo. CONCLUSIONS: t-PA, in addition to its known role in invasion, plays other critical roles in pancreas tumor progression, stimulating cancer cell proliferation and tumor-associated angiogenesis. PMID- 11875016 TI - Surveying the case for surveillance. PMID- 11875017 TI - (Post) inflammatory visceral hyperalgesia: don't we believe what we don't see? PMID- 11875018 TI - The NOD2 gene and Crohn's disease: another triumph for molecular genetics. PMID- 11875019 TI - Fantastic voyage through the gut. PMID- 11875020 TI - Hereditary diffuse gastric cancer: more answers or more questions? PMID- 11875021 TI - Prevention of varices rebleeding: are drugs better after all? PMID- 11875023 TI - Identification of multiple genetic loci linked to the propensity for "behavioral despair" in mice. AB - The forced swim test (FST) and tail suspension test (TST) are widely used and well established screening paradigms for antidepressants. A variety of antidepressive agents are known to reduce immobility time in both FST and TST. To identify genetic determinants of immobility duration in both tests, we analyzed 560 F2 mice from an intercross between C57BL/6 (B6) and C3H/He (C3) strains. Composite interval mapping revealed five major loci (suggestive and significant linkage) affecting immobility in the FST, and four loci for the TST. The quantitative trait loci (QTL) on chromosomes 8 and 11 overlap between the two behavioral measures. Genome-wide interaction analysis, which was developed to identify locus pairs that may contribute epistatically to a phenotype, detected two pairs of chromosomal loci for the TST. The QTL on chromosome 11 and its associated epistatic TST-QTL on chromosome X encode gamma-aminobutyric acid type A (GABA(A)) receptor subunits as candidates. Sequence and expression analyses of these genes from the two parental strains revealed a significantly lower expression of the alpha1 subunit gene in the frontal cortex of B6 mice compared to C3 mice. The present quantitative trait study should open up avenues for identifying novel molecular targets for antidepressants and unraveling the complex genetic mechanisms of depressive and anxiety disorders. PMID- 11875024 TI - Exploration of novel motifs derived from mouse cDNA sequences. AB - We performed a systematic maximum density subgraph (MDS) detection of conserved sequence regions to discover new, biologically relevant motifs from a set of 21,050 conceptually translated mouse cDNA (FANTOM1) sequences. A total of 3202 candidate sequences, which shared similar regions over >20 amino acid residues, were screened against known conserved regions listed in Pfam, ProDom, and InterPro. The filtering procedure resulted in 139 FANTOM1 sequences belonging to 49 new motif candidates. Using annotations and multiple sequence alignment information, we removed by visual inspection 42 candidates whose members were found to be false positives because of sequence redundancy, alternative splicing, low complexity, transcribed retroviral repeat elements contained in the region of the predicted open reading frame, and reports in the literature. The remaining seven motifs have been expanded by hidden Markov model (HMM) profile searches of SWISS-PROT/TrEMBL from 28 FANTOM1 sequences to 164 members and analyzed in detail on sequence and structure level to elucidate the possible functions of motifs and members. The novel and conserved motif MDS00105 is specific for the mammalian inhibitor of growth (ING) family. Three submotifs MDS00105.1-3 are specific for ING1/ING1L, ING1-homolog, and ING3 subfamilies. The motif MDS00105 together with a PHD finger domain constitutes a module for ING proteins. Structural motif MDS00113 represents a leucine zipper-like motif. Conserved motif MDS00145 is a novel 1-acyl-SN-glycerol-3-phosphate acyltransferase (AGPAT) submotif containing a transmembrane domain that distinguishes AGPAT3 and AGPAT4 from all other acyltransferase domain-containing proteins. Functional motif MDS00148 overlaps with the kazal-type serine protease inhibitor domain but has been detected only in an extracellular loop region of solute carrier 21 (SLC21) (organic anion transporters) family members, which may regulate the specificity of anion uptake. Our motif discovery not only aided in the functional characterization of new mouse orthologs for potential drug targets but also allowed us to predict that at least 16 other new motifs are waiting to be discovered from the current SWISS PROT/TrEMBL database. PMID- 11875025 TI - The human ribosomal protein genes: sequencing and comparative analysis of 73 genes. AB - The ribosome, as a catalyst for protein synthesis, is universal and essential for all organisms. Here we describe the structure of the genes encoding human ribosomal proteins (RPs) and compare this class of genes among several eukaryotes. Using genomic and full-length cDNA sequences, we characterized 73 RP genes and found that (1) transcription starts at a C residue within a characteristic oligopyrimidine tract; (2) the promoter region is GC rich, but often has a TATA box or similar sequence element; (3) the genes are small (4.4 kb), but have as many as 5.6 exons on average; (4) the initiator ATG is in the first or second exon and is within plus minus 5 bp of the first intron boundaries in about half of cases; and (5) 5'- and 3'-UTRs are significantly smaller (42 bp and 56 bp, respectively) than the genome average. Comparison of RP genes from humans, Drosophila melanogaster, Caenorhabditis elegans, and Saccharomyces cerevisiae revealed the coding sequences to be highly conserved (63% homology on average), although gene size and the number of exons vary. The positions of the introns are also conserved among these species as follows: 44% of human introns are present at the same position in either D. melanogaster or C. elegans, suggesting RP genes are highly suitable for studying the evolution of introns. PMID- 11875026 TI - Processed pseudogenes of human endogenous retroviruses generated by LINEs: their integration, stability, and distribution. AB - We report here the presence of numerous processed pseudogenes derived from the W family of endogenous retroviruses in the human genome. These pseudogenes are structurally colinear with the retroviral mRNA followed by a poly(A) tail. Our analysis of insertion sites of HERV-W processed pseudogenes shows a strong preference for the insertion motif of long interspersed nuclear element (LINE) retrotransposons. The genomic distribution, stability during evolution, and frequent truncations at the 5' end resemble those of the pseudogenes generated by LINEs. We therefore suggest that HERV-W processed pseudogenes arose by multiple and independent LINE-mediated retrotransposition of retroviral mRNA. These data document that the majority of HERV-W copies are actually nontranscribed promoterless pseudogenes. The current search for HERV-Ws associated with several human diseases should concentrate on a small subset of transcriptionally competent elements. PMID- 11875027 TI - Recombination rate and the distribution of transposable elements in the Drosophila melanogaster genome. AB - We analyzed the distribution of 54 families of transposable elements (TEs; transposons, LTR retrotransposons, and non-LTR retrotransposons) in the chromosomes of Drosophila melanogaster, using data from the sequenced genome. The density of LTR and non-LTR retrotransposons (RNA-based elements) was high in regions with low recombination rates, but there was no clear tendency to parallel the recombination rate. However, the density of transposons (DNA-based elements) was significantly negatively correlated with recombination rate. The accumulation of TEs in regions of reduced recombination rate is compatible with selection acting against TEs, as selection is expected to be weaker in regions with lower recombination. The differences in the relationship between recombination rate and TE density that exist between chromosome arms suggest that TE distribution depends on specific characteristics of the chromosomes (chromatin structure, distribution of other sequences), the TEs themselves (transposition mechanism), and the species (reproductive system, effective population size, etc.), that have differing influences on the effect of natural selection acting against the TE insertions. PMID- 11875028 TI - Domain-level differences in microsatellite distribution and content result from different relative rates of insertion and deletion mutations. AB - Microsatellites (short tandem polynucleotide repeats) are found throughout eukaryotic genomes at frequencies many orders of magnitude higher than the frequencies predicted to occur by chance. Most of these microsatellites appear to have evolved in a generally neutral manner. In contrast, microsatellites are generally absent from bacterial genomes except in locations where they provide adaptive functional variability, and these appear to have evolved under selection. We demonstrate a mutational bias towards deletion (repeat contraction) in a native chromosomal microsatellite of the bacterium Mycoplasma gallisepticum, through the collection and analysis of independent mutations in the absence of natural selection. Using this and similar existing data from two other bacterial species and four eukaryotic species, we find strong evidence that deletion biases resulting in repeat contraction are common in bacteria, while eukaryotic microsatellites generally experience unbiased mutation or a bias towards insertion (repeat expansion). This difference in mutational bias suggests that eukaryotic microsatellites should generally expand wherever selection does not exclude them, whereas bacterial microsatellites should be driven to extinction by mutational pressure wherever they are not maintained by selection. This is consistent with observed bacterial and eukaryotic microsatellite distributions. Hence, mutational biases that differ between eukaryotes and bacteria can account for many of the observed differences in microsatellite DNA content and distribution found in these two groups of organisms. PMID- 11875029 TI - Genomic microsatellites as evolutionary chronometers: a test in wild cats. AB - Nuclear microsatellite loci (2- to 5-bp tandem repeats) would seem to be ideal markers for population genetic monitoring because of their abundant polymorphism, wide dispersal in vertebrate genomes, near selective neutrality, and ease of assessment; however, questions about their mode of generation, mutation rates and ascertainment bias have limited interpretation considerably. We have assessed the patterns of genomic diversity for ninety feline microsatellite loci among previously characterized populations of cheetahs, lions and pumas in recapitulating demographic history. The results imply that the microsatellite diversity measures (heterozygosity, allele reconstitution and microsatellite allele variance) offer proportionate indicators, albeit with large variance, of historic population bottlenecks and founder effects. The observed rate of reconstruction of new alleles plus the growth in the breadth of microsatellite allele size (variance) was used here to develop genomic estimates of time intervals following historic founder events in cheetahs (12,000 yr ago), in North American pumas (10,000-17,000 yr ago), and in Asiatic lions of the Gir Forest (1000-4000 yr ago). PMID- 11875030 TI - Computational comparison of human genomic sequence assemblies for a region of chromosome 4. AB - Much of the available human genomic sequence data exist in a fragmentary draft state following the completion of the initial high-volume sequencing performed by the International Human Genome Sequencing Consortium (IHGSC) and Celera Genomics (CG). We compared six draft genome assemblies over a region of chromosome 4p (D4S394-D4S403), two consecutive releases by the IHGSC at University of California, Santa Cruz (UCSC), two consecutive releases from the National Centre for Biotechnology Information (NCBI), the public release from CG, and a hybrid assembly we have produced using IHGSC and CG sequence data. This region presents particular problems for genomic sequence assembly algorithms as it contains a large tandem repeat and is sparsely covered by draft sequences. The six assemblies differed both in terms of their relative coverage of sequence data from the region and in their estimated rates of misassembly. The CG assembly method attained the lowest level of misassembly, whereas NCBI and UCSC assemblies had the highest levels of coverage. All assemblies examined included <60% of the publicly available sequence from the region. At least 6% of the sequence data within the CG assembly for the D4S394-D4S403 region was not present in publicly available sequence data. We also show that even in a problematic region, existing software tools can be used with high-quality mapping data to produce genomic sequence contigs with a low rate of rearrangements. PMID- 11875031 TI - A tale of two genotypes: consistency between two high-throughput genotyping centers. AB - Multiple genome-wide scans involving sib-pairs or limited pedigrees have been extensively used for a wide number of complex genetic conditions. Comparing data from two or more scans, as well as combining data, require an understanding of the sources of genotyping errors and data discrepancies. We have conducted two genome-wide scans for age-related maculopathy using the Center for Inherited Disease Research (CIDR) and the Mammalian Genotyping Service (MGS). Thirty individuals were typed in common, in order to allow for the alignment of alleles and comparison of the data sets. The analysis of these 8914 genotypes distributed over 321 markers in common demonstrated excellent agreement between these two laboratories, which have low rates of internal errors. Under the assumption that within each genotype, the smaller MGS allele should correspond to the smaller CIDR allele, the alleles align well between the two centers, with only a small fraction (less than 0.65%) of the aligned alleles showing large differences in sizes. However, since called allele sizes are integer "labels" which may not directly reflect the true underlying allele sizes, it is important to carefully prepare in advance if one wishes to merge data from different laboratories. In particular, it would not suffice to attempt to align alleles by typing only one or two controls in common. Fortunately, for the purposes of linkage analysis, one can avoid merging difficulties by simply carrying out linkage analyses using laboratory-specific allele labels and allele frequencies for each laboratory specific subset of the data. PMID- 11875032 TI - Accounting for human polymorphisms predicted to affect protein function. AB - A major interest in human genetics is to determine whether a nonsynonymous single base nucleotide polymorphism (nsSNP) in a gene affects its protein product and, consequently, impacts the carrier's health. We used the SIFT (Sorting Intolerant From Tolerant) program to predict that 25% of 3084 nsSNPs from dbSNP, a public SNP database, would affect protein function. Some of the nsSNPs predicted to affect function were variants known to be associated with disease. Others were artifacts of SNP discovery. Two reports have indicated that there are thousands of damaging nsSNPs in an individual's human genome; we find the number is likely to be much lower. PMID- 11875033 TI - Oligonucleotide arrays for high-throughput SNPs detection in the MHC class I genes: HLA-B as a model system. AB - A simple and efficient oligonucleotide array was developed to identify single nucleotide polymorphisms (SNPs) encoded within the highly polymorphic human major histocompatibility complex (MHC) using HLA-B as a model system. A total of 137 probes were designed to represent all known polymorphisms encoded in exons 2 and 3. PCR products were amplified from human genomic DNA and allowed to hybridize with the oligonucleotide array. Hybridization was detected by fluorescence scanning, and HLA-B alleles were assigned by quantitative analysis of the hybridization results. Variables known to influence the specificity of hybridization, such as oligonucleotide probe size, spacer length, surface density, hybridization conditions, and array uniformity and stability were studied. The efficiency and specificity of identifying HLA-B SNPs using the oligonucleotide arrays was evaluated by blinded analysis of 100 samples from unrelated individuals representing all HLA-B phenotypes. The oligonucleotide array method described in this paper provides unambiguous detection of complex heterozygous SNP combinations. This methodological approach may be applied to other highly polymorphic gene systems. PMID- 11875034 TI - Computational detection and location of transcription start sites in mammalian genomic DNA. AB - Transcription, the process whereby RNA copies are made from sections of the DNA genome, is directed by promoter regions. These define the transcription start site, and also the set of cellular conditions under which the promoter is active. At least in more complex species, it appears to be common for genes to have several different transcription start sites, which may be active under different conditions. Eukaryotic promoters are complex and fairly diffuse structures, which have proven hard to detect in silico. We show that a novel hybrid machine learning method is able to build useful models of promoters for >50% of human transcription start sites. We estimate specificity to be >70%, and demonstrate good positional accuracy. Based on the structure of our learned models, we conclude that a signal resembling the well known TATA box, together with flanking regions of C-G enrichment, are the most important sequence-based signals marking sites of transcriptional initiation at a large class of typical promoters. PMID- 11875035 TI - Consensus promoter identification in the human genome utilizing expressed gene markers and gene modeling. AB - Deciphering the human genome includes locating the promoters that initiate transcription and identifying the exons of genes. Many promoter prediction programs have been proposed, but when they are applied to extended regions of the genome, most of their predictions are false-positives. The extensive collection of gene transcript sequences is an important new source of information, which has not been used previously in promoter predictions. Our approach is to enhance the specificity of predictions by restricting the genomic regions that are searched using gene transcript alignments as anchors in the genome for gene modeling. We developed a consensus promoter prediction method combining previously developed algorithms with the GENSCAN gene modeling program. Our method, CONPRO (CONsensus PROmoter), identifies promoters with very high confidence, and the predicted promoters are guaranteed to be associated with genes. On our test data set, the method correctly detects promoters for approximately half of all human genes (37% 71%), and most predictions are true promoters (85%-90%). Applying our method to the human genome and human genes from the Unigene data set, we find the promoters for 13,744 genes. Of these, 6440 are genes with a functionally cloned mRNA, and 7304 are novel genes for which only expressed sequence tags (ESTs) are available. Candidate promoters for many novel genes will be a useful resource in elucidating complex biological response mechanisms. PMID- 11875036 TI - Extraction of functional binding sites from unique regulatory regions: the Drosophila early developmental enhancers. AB - The early developmental enhancers of Drosophila melanogaster comprise one of the most sophisticated regulatory systems in higher eukaryotes. An elaborate code in their DNA sequence translates both maternal and early embryonic regulatory signals into spatial distribution of transcription factors. One of the most striking features of this code is the redundancy of binding sites for these transcription factors (BSTF). Using this redundancy, we explored the possibility of predicting functional binding sites in a single enhancer region without any prior consensus/matrix description or evolutionary sequence comparisons. We developed a conceptually simple algorithm, Scanseq, that employs an original statistical evaluation for identifying the most redundant motifs and locates the position of potential BSTF in a given regulatory region. To estimate the biological relevance of our predictions, we built thorough literature-based annotations for the best-known Drosophila developmental enhancers and we generated detailed distribution maps for the most robust binding sites. The high statistical correlation between the location of BSTF in these experiment-based maps and the location predicted in silico by Scanseq confirmed the relevance of our approach. We also discuss the definition of true binding sites and the possible biological principles that govern patterning of regulatory regions and the distribution of transcriptional signals. PMID- 11875037 TI - High-density cell microarrays for parallel functional determinations. AB - Whole-genome sequencing projects have generated a wealth of gene sequences from a variety of organisms. A major challenge is to rapidly uncover gene regulatory circuits and their functional manifestations at the cellular level. Here we report the coupled fabrication of nanocraters ranging in size from 100 pL to 1.5 nL on permeable membranes for culturing cells. Using this approach, we developed bacterial and yeast cell microarrays that allowed phenotypic determinations of gene activities and drug targets on a large scale. Cell microarrays will therefore be a particularly useful tool for studying phenotypes of gene activities on a genome-wide scale. PMID- 11875038 TI - Novel fluorescence labeling and high-throughput assay technologies for in vitro analysis of protein interactions. AB - We developed and tested a simple method for fluorescence labeling and interaction analysis of proteins based on a highly efficient in vitro translation system combined with high-throughput technologies such as microarrays and fluorescence cross-correlation spectroscopy (FCCS). By use of puromycin analogs linked to various fluorophores through a deoxycytidylic acid linker, a single fluorophore can be efficiently incorporated into a protein at the carboxyl terminus during in vitro translation. We confirmed that the resulting fluorescently labeled proteins are useful for probing protein-protein and protein-DNA interactions by means of pulldown assay, DNA microarrays, and FCCS in model experiments. These fluorescence assay systems can be easily extended to highly parallel analysis of protein interactions in studies of functional genomics. PMID- 11875039 TI - Cross-referencing eukaryotic genomes: TIGR Orthologous Gene Alignments (TOGA). AB - Comparative genomics promises to rapidly accelerate the identification and functional classification of biologically important human genes. We developed the TIGR Orthologous Gene Alignment (TOGA; ) database to provide a cross-reference between fully and partially sequenced eukaryotic transcribed sequences. Starting with the assembled expressed sequence tag (EST) and gene sequences that comprise the 28 TIGR Gene Indices, we used high stringency pair-wise sequence searches and a reflexive, transitive closure process to associate sequence-specific best hits, generating 32,652 tentative ortholog groups (TOGs). This has allowed us to identify putative orthologs and paralogs for known genes, as well as those that exist only as uncharacterized ESTs and to provide links to additional information including genome sequence and mapping data. TOGA provides an important new resource for the analysis of gene function in eukaryotes. In addition, an analysis of the most widely represented sequences can begin to provide insight into eukaryotic biological processes. PMID- 11875040 TI - Gene3D: structural assignment for whole genes and genomes using the CATH domain structure database. AB - We present a novel web-based resource, Gene3D, of precalculated structural assignments to gene sequences and whole genomes. This resource assigns structural domains from the CATH database to whole genes and links these to their curated functional and structural annotations within the CATH domain structure database, the functional Dictionary of Homologous Superfamilies (DHS) and PDBsum. Currently Gene3D provides annotation for 36 complete genomes (two eukaryotes, six archaea, and 28 bacteria). On average, between 30% and 40% of the genes of a given genome can be structurally annotated. Matches to structural domains are found using the profile-based method (PSI-BLAST). and a novel protocol, DRange, is used to resolve conflicts in matches involving different homologous superfamilies. PMID- 11875041 TI - BAC library of T. pallidum DNA in E. coli. AB - Treponema pallidum subspecies pallidum (Nichols) chromosomal DNA was used to construct a large insert bacterial artificial chromosome (BAC) library in Escherichia coli DH10B using the pBeloBAC11 cloning vector; 678 individual insert termini of 339 BAC clones (13.9 x coverage) were sequenced and the cloned chromosomal region in each clone was determined by comparison to the genomic sequence. A single 15.6-kb region of the T. pallidum chromosome was missing in the BAC library, between bp 248727 and 264323. In addition to the 12 open reading frames (ORFs) coded by this region, one additional ORF (TP0596) was not cloned as an intact gene. Altogether, 13 predicted T. pallidum ORFs (1.25% of the total) were incomplete or missing in the library. Three of 338 clones mapped by restriction enzyme digestion had detectable deletions and one clone had a detectable insertion within the insert. Of mapped clones, 19 were selected to represent the minimal set of E. coli BAC clones covering 1026 of the total 1040 (98.7%) predicted T. pallidum ORFs. Using this minimal set of clones, at least 12 T. pallidum proteins were shown to react with pooled sera from rabbits immunized with T. pallidum, indicating that at least some T. pallidum genes are transcribed and expressed in E. coli. PMID- 11875042 TI - Human deafness dystonia syndrome is caused by a defect in assembly of the DDP1/TIMM8a-TIMM13 complex. AB - Mohr-Tranebjaerg syndrome (MTS/DFN-1) or deafness/dystonia syndrome results from a mutation in deafness/dystonia protein 1/translocase of mitochondrial inner membrane 8a (DDP1/TIMM8a). DDP1/TIMM8a is similar to a family of yeast proteins in the mitochondrial intermembrane space which mediate the import and insertion of inner membrane proteins. We now show that TIMM8a assembles in a 70 kDa complex in the intermembrane space with TIMM13. DDP1/TIMM8a is not detectable in fibroblasts derived from a patient with a missense mutation in the DDP1/TIMM8a gene; the point mutation results in cysteine-66 being changed to tryptophan-66 in the conserved 'twin CX(3)C' motif. The corresponding mutation in yeast translocase of inner membrane 8p (Tim8p) yields an unstable protein that does not assemble with yeast Tim13p. DDP1/TIMM8a, when expressed with TIMM13 in yeast mitochondria lacking the Tim8p-Tim13p complex, restores Tim23p import, and TIMM8a and TIMM13 can be cross-linked to the hTim23 import intermediate in rat and yeast mitochondria. In a similar manner to Tim8p, TIMM8a seemingly mediates the import of hTim23. Deafness/dystonia syndrome thus may be caused by decreased levels of Tim23 in the mitochondrial inner membrane in affected tissues. PMID- 11875043 TI - Knockout mouse model for Fxr2: a model for mental retardation. AB - Fragile X syndrome is a common form of mental retardation caused by the absence of the FMR1 protein, FMRP. Fmr1 knockout mice exhibit a phenotype with some similarities to humans, such as macro-orchidism and behavioral abnormalities. Two homologs of FMRP have been identified, FXR1P and FXR2P. These proteins show high sequence similarity, including all functional domains identified in FMRP, such as RNA binding domains. They have an overlap in tissue distribution to that of FMRP. Interactions between the three FXR proteins have also been described. FXR2P shows high expression in brain and testis, like FMRP. To study the function of FXR2P, we generated an Fxr2 knockout mouse model. No pathological differences between knockout and wild-type mice were found in brain or testis. Given the behavioral phenotype in fragile X patients and the phenotype previously reported for the Fmr1 knockout mouse, we performed a thorough evaluation of the Fxr2 knockout phenotype using a behavioral test battery. Fxr2 knockout mice were hyperactive (i.e. traveled a greater distance, spent more time moving and moved faster) in the open-field test, impaired on the rotarod test, had reduced levels of prepulse inhibition, displayed less contextual conditioned fear, impaired at locating the hidden platform in the Morris water task and were less sensitive to a heat stimulus. Interestingly, there are some behavioral phenotypes in Fxr2 knockout mice which are similar to those observed in Fmr1 knockout mice, but there are also some different behavioral abnormalities that are only observed in the Fxr2 mutant mice. The findings implicate a role for Fxr2 in central nervous system function. PMID- 11875044 TI - Late onset neurological phenotype of the X-ALD gene inactivation in mice: a mouse model for adrenomyeloneuropathy. AB - Adrenomyeloneuropathy (AMN) and cerebral childhood adrenoleukodystrophy (CCALD) are the main phenotypic variants of an X-linked inherited metabolic disorder causing demyelination, X-linked adrenoleukodystrophy (X-ALD). It is caused by mutations in the ABCD1 (ALD) gene encoding a peroxisomal ABC transporter. Inactivation of the murine ALD gene does not lead to a detectable clinical phenotype in mice up to 6 months, and no cerebral pathology resembling the childhood form (CCALD) was observed. In this work, we show that older ALD deficient mice exhibit an abnormal neurological and behavioral phenotype, starting at around 15 months. This is correlated with slower nerve conduction, and with myelin and axonal anomalies detectable in the spinal cord and sciatic nerve, but not in brain. The phenotype of ALD-deficient mice mimics features of human AMN, thus providing a model for investigating the pathogenesis of this disease. PMID- 11875045 TI - A Gja8 (Cx50) point mutation causes an alteration of alpha 3 connexin (Cx46) in semi-dominant cataracts of Lop10 mice. AB - Mutations of connexin alpha 8 (GJA8 or Cx50) and connexin alpha 3 (GJA3 or Cx46) in humans have been reported to cause cataracts with semi-dominant inheritance patterns. Targeted null mutations in Gja8 and Gja3 in mice cause cataracts with recessive inheritance. The molecular bases for these differences in inheritance patterns and the mechanism for cataractogenesis in these mutants are poorly understood. We recently mapped an autosomal semi-dominant cataract [lens opacity 10 (Lop10)] mutation to mouse chromosome 3 and identified a missense mutation (G- >C) in the Gja8 gene, which causes glycine at codon 22 to be replaced with arginine (G22R). Moreover, we demonstrated that the alpha 8 G22R isoform is a loss-of-function mutant for alpha 8, as well as a dominant mutation for reducing the phosphorylated forms of alpha 3 connexin in vivo. To test the hypothesis that the alteration of endogenous alpha 3 connexin in Lop10 mice led to a unique lens phenotype, we generated double mutant offspring between Lop10 and the Gja3(tm1) (alpha 3(-/-)) mice. The double homozygous mutant mice (Lop10/Lop10 alpha 3(-/-)) showed relatively normal lens cortical fibers compared to the Lop10 mice. A functional impairment of endogenous alpha 3 connexin is therefore partly responsible for cellular phenotypes in the Lop10 mice. This study has provided some novel molecular insights into mouse and human cataractogenesis caused by alpha 8 and alpha 3 mutations. These mouse models will be useful for investigating the mechanistic relationship between gap junction impairment and cataract formation. PMID- 11875046 TI - Molecular chaperones enhance the degradation of expanded polyglutamine repeat androgen receptor in a cellular model of spinal and bulbar muscular atrophy. AB - Spinal and bulbar muscular atrophy (SBMA) is one of a growing number of neurodegenerative diseases caused by a polyglutamine-encoding CAG trinucleotide repeat expansion, and is caused by an expansion within exon 1 of the androgen receptor (AR) gene. The family of polyglutamine diseases is characterized by the presence of ubiquitinated, intranuclear inclusions associated with molecular chaperones and 26S proteasome components, although the role of these inclusions in the pathogenesis of polyglutamine diseases remains unclear. The over expression of molecular chaperones of the Hsp70 and Hsp40 families has been shown to modulate inclusion frequency and cellular toxicity. We developed a cell culture system which enables the quantitative analysis of the effects of molecular chaperones on the biochemical properties of an expanded repeat AR. Using this approach, we demonstrate that Hsp70 and its co-chaperone Hsp40 not only increase expanded repeat AR solubility, but function to enhance the degradation of expanded repeat AR through the proteasome. Furthermore, our studies indicate that these molecular chaperones significantly decrease the half life of an expanded repeat AR. Molecular chaperone enhancement of protein degradation points to the modulation of molecular chaperones as a potential therapeutic target for polyglutamine diseases. PMID- 11875047 TI - A mouse model of TSC1 reveals sex-dependent lethality from liver hemangiomas, and up-regulation of p70S6 kinase activity in Tsc1 null cells. AB - Tuberous sclerosis (TSC) is a autosomal dominant genetic disorder caused by mutations in either TSC1 or TSC2, and characterized by benign hamartoma growth. We developed a murine model of Tsc1 disease by gene targeting. Tsc1 null embryos die at mid-gestation from a failure of liver development. Tsc1 heterozygotes develop kidney cystadenomas and liver hemangiomas at high frequency, but the incidence of kidney tumors is somewhat lower than in Tsc2 heterozygote mice. Liver hemangiomas were more common, more severe and caused higher mortality in female than in male Tsc1 heterozygotes. Tsc1 null embryo fibroblast lines have persistent phosphorylation of the p70S6K (S6K) and its substrate S6, that is sensitive to treatment with rapamycin, indicating constitutive activation of the mTOR-S6K pathway due to loss of the Tsc1 protein, hamartin. Hyperphosphorylation of S6 is also seen in kidney tumors in the heterozygote mice, suggesting that inhibition of this pathway may have benefit in control of TSC hamartomas. PMID- 11875049 TI - Identification of an IMPDH1 mutation in autosomal dominant retinitis pigmentosa (RP10) revealed following comparative microarray analysis of transcripts derived from retinas of wild-type and Rho(-/-) mice. AB - Comparative analysis of the transcriptional profiles of approximately 6000 genes in the retinas of wild-type mice with those carrying a targeted disruption of the rhodopsin gene was undertaken by microarray analysis. This revealed a series of transcripts, of which some were derived from genes known to map at retinopathy loci, levels of which were reduced or elevated in the retinas of Rho(-/-) mice lacking functional photoreceptors. The human homologue of one of these genes, encoding inosine monophosphate dehydrogenase type 1 (IMPDH1), maps to the region of 7q to which an adRP gene (RP10) had previously been localized. Mutational screening of DNA from the Spanish adRP family, originally used to localize the RP10 gene, revealed an Arg224Pro substitution co-segregating with the disease phenotype. The amino acid at position 224 of the IMPDH1 protein is conserved among species and the substitution is not present in healthy, unrelated individuals of European origin. These data provide strong evidence that mutations within the IMPDH1 gene cause adRP, and validate approaches to mutation detection involving comparative analysis of global transcription profiles in normal and degenerating retinal tissues. Other genes showing significant alterations in expression include some with anti-apoptotic functions and many encoding components of the extracellular matrix or cytoskeleton, a possible reflection of a response by Muller cells to preserve the remaining outer nuclear layer of the retina. We suggest that those genes identified are prime candidates for etiological involvement in degenerative retinal disease. PMID- 11875050 TI - Mutations in the inosine monophosphate dehydrogenase 1 gene (IMPDH1) cause the RP10 form of autosomal dominant retinitis pigmentosa. AB - Autosomal dominant retinitis pigmentosa (adRP) is a heterogeneous set of progressive retinopathies caused by several distinct genes. One locus, the RP10 form of adRP, maps to human chromosome 7q31.1 and may account for 5-10% of adRP cases among Americans and Europeans. We identified two American families with the RP10 form of adRP by linkage mapping and used these families to reduce the linkage interval to 3.45 Mb between the flanking markers D7S686 and RP-STR8. Sequence and transcript analysis identified 54 independent genes within this region, at least 10 of which are retinal-expressed and thus candidates for the RP10 gene. A screen of retinal transcripts comparing retinas from normal mice to retinas from crx-/crx- knockout mice (with poorly differentiated photoreceptors) demonstrated a 6-fold reduction in one candidate, inosine monophosphate dehydrogenase 1 (IMPDH1; EC 1.1.1.205). Since many of the genes known to cause retinitis pigmentosa are under CRX control in photoreceptors, IMPDH1 became a high-priority candidate for mutation screening. DNA sequencing of affected individuals from the two American RP10 families revealed a GAC-->AAC transition in codon 226 substituting an asparagine for an aspartic acid in both families. The identical mutation was also found in a British RP10 family. The Asp226Asn missense mutation is present in all affected individuals tested and absent from unaffected controls. The aspartic acid at codon 226 is conserved in all IMPDH genes, in all species examined, including bacteria, suggesting that this mutation is highly deleterious. Subsequent screening of probands from 60 other adRP families revealed an additional family with this mutation, confirming its association with retinitis pigmentosa and the relatively high frequency of this mutation. Another IMPDH1 substitution, Val268Ile, was also observed in this cohort of patients but not in controls. IMPDH1 is a ubiquitously expressed enzyme, functioning as a homotetramer, which catalyzed the rate-limiting step in de novo synthesis of guanine nucleotides. As such, it plays an important role in cyclic nucleoside metabolism within photoreceptors. Several classes of drugs are known to affect IMPDH isoenzymes, including nucleotide and NAD analogs, suggesting that small-molecule therapy may be available, one day, for RP10 patients. PMID- 11875051 TI - The role of matrix metalloproteinase polymorphisms in the rate of decline in lung function. AB - The matrix metalloproteinases (MMPs) comprise a family of at least 20 proteolytic enzymes that play an essential role in tissue remodeling. MMP1 (interstitial collagenase), MMP9 (gelatinase B) and MMP12 (macrophage elastase) are thought to be important in the development of emphysema. A number of naturally occurring polymorphisms of human MMP gene promoters have been identified and found to alter transcriptional activity. Additionally, we detected a novel polymorphism in the MMP12 coding region (Asn357Ser). The aim of this study was to investigate the role of MMP polymorphisms in the development of chronic obstructive lung disease. We determined the prevalence of these polymorphisms in 590 continuing smokers chosen from the National Heart Lung and Blood Institute, Lung Health Study for having the fastest (n = 284) and slowest (n = 306) 5 year rate of decline of lung function. Of the five polymorphisms, only G-1607GG was associated with a rate of decline in lung function. The -1607GG allele was associated with a fast rate of decline (P = 0.02) [corrected]. However, haplotypes consisting of alleles from the MMP1 G-1607GG and MMP12 Asn357Ser polymorphisms were associated with rate of decline of lung function (P = 0.0007). These data suggest that polymorphisms in the MMP1 and MMP12 genes, but not MMP9, are either causative factors in smoking related lung injury or are in linkage disequilibrium with causative polymorphisms. PMID- 11875048 TI - Different evolutionary processes shaped the mouse and human olfactory receptor gene families. AB - We report a comprehensive comparative analysis of human and mouse olfactory receptor (OR) genes. The OR family is the largest mammalian gene family known. We identify approximately 93% of an estimated 1500 mouse ORs, exceeding previous estimates and the number of human ORs by 50%. Only 20% are pseudogenes, giving a functional OR repertoire in mice that is three times larger than that of human. The proteins encoded by intact human ORs are less highly conserved than those of mouse, in patterns that suggest that even some apparently intact human OR genes may encode non-functional proteins. Mouse ORs are clustered in 46 genomic locations, compared to a much more dispersed pattern in human. We find orthologous clusters at syntenic human locations for most mouse genes, indicating that most OR gene clusters predate primate-rodent divergence. However, many recent local OR duplications in both genomes obscure one-to-one orthologous relationships, thereby complicating cross-species inferences about OR-ligand interactions. Local duplications are the major force shaping the gene family. Recent interchromosomal duplications of ORs have also occurred, but much more frequently in human than in mouse. In addition to clarifying the evolutionary forces shaping this gene family, our study provides the basis for functional studies of the transcriptional regulation and ligand-binding capabilities of the OR gene family. PMID- 11875052 TI - SRp30c-dependent stimulation of survival motor neuron (SMN) exon 7 inclusion is facilitated by a direct interaction with hTra2 beta 1. AB - Proximal spinal muscular atrophy (SMA) is caused by the homozygous loss of survival motor neuron (SMN1). SMN2, a nearly identical copy gene, is present in all SMA patients; however this gene cannot provide protection from disease due to the aberrant splicing of a critical exon. SMN1-derived transcripts are exclusively full-length, whereas SMN2-derived transcripts predominantly lack SMN exon 7. A single non-polymorphic nucleotide difference (C in SMN1; T in SMN2) is responsible for the alternative splicing patterns. We have previously shown that transient expression of an SR-like splicing factor, hTra2 beta 1, stimulates inclusion of exon 7 in SMN2-derived mini-gene transcripts through an interaction with the AG-rich exonic splice enhancer within exon 7. We now demonstrate that a second splicing factor, SRp30c, can stimulate SMN exon 7-inclusion and that this activity required the same AG-rich enhancer as hTra2 beta 1. SRp30c did not directly associate with SMN exon 7; rather its association with the exonic enhancer was mediated by a direct interaction with hTra2 beta 1. In the absence of the hTra2 beta 1 binding site, SRp30c failed to complex with SMN exon 7. Taken together, these results identify SRp30c as a modulator of SMN exon 7-inclusion and provide insight into the molecular regulation of this critical exon. PMID- 11875053 TI - Coding haplotype analysis supports HCR as the putative susceptibility gene for psoriasis at the MHC PSORS1 locus. AB - PSORS1, near HLA-C, is the major genetic determinant of psoriasis. We present genetic and structural evidence suggesting a major role for the HCR gene at the PSORS1 locus. Genotyping of 419 families from six populations revealed that coding single-nucleotide polymorphisms of HCR formed a conserved allele HCR*WWCC that associated highly significantly with psoriasis and with the HLA-Cw6 allele in all populations. Because of strong linkage disequilibrium between HLA-Cw6 and HCR*WWCC, the two genes could not be genetically distinguished by this sample size. However, the variant HCR allele was predicted to differ in secondary structure from the wild-type protein. HCR protein expression in lesional psoriatic skin differed considerably from that observed in normal skin. These results provide strong evidence for the HCR*WWCC allele as a major genetic determinant for psoriasis, probably by a mechanism impacting on keratinocyte proliferation. PMID- 11875054 TI - A frequent mild mutation in ALG6 may exacerbate the clinical severity of patients with congenital disorder of glycosylation Ia (CDG-Ia) caused by phosphomannomutase deficiency. AB - Single nucleotide polymorphisms occur throughout the human genome. A gene that causes one of the congenital disorders of glycosylation (CDG) has a mutation (911T-->C ) that changes a phenylalanine to serine at position 304 (F304S) of the alpha 1,3 glucosyl transferase. We show that this change reduces the ability of the gene product to rescue defective glycosylation of an alg6-deficient strain of Saccharomyces cerevisiae during rapid growth. This finding suggested that the mutation might affect glycosylation in humans. We therefore compared the frequency of this variant in 301 controls and in 101 CDG patients who carry known mutations in other genes involved in CDG, i.e. PMM2 (CDG-Ia; 91 patients) and MPI (CDG-Ib; 10 patients). The variant allele frequency is identical in both CDG patients (0.30) and controls (0.28). Importantly, the F304S genotype frequency in 55 CDG-Ia patients classified as mild/moderate (n = 28), or severe (n = 27) was significantly higher in severely affected patients (0.41) than in mild/moderately affected patients (0.21). Mortality (n = 9) was higher when F304S was present (n = 6). Severely affected patients with the PMM2 mutations F119L/R141H (n = 22) carry the F304S mutation more often (0.36) than mildly affected patients (0.18, n = 11) with this mutation. Clinical severity of mildly affected sibs with the same PMM2 mutations did not correlate with F304S genotype. Thus, the presence of the F304S allele may exacerbate the clinical outcome, especially in severely affected CDG patients. We speculate that this type of variant may be implicated in other multi-factorial disorders that involve N-glycosylation. PMID- 11875055 TI - Mutations in the RPGR gene cause X-linked cone dystrophy. AB - X-linked cone dystrophy is a type of hereditary retinal degeneration characterized by a progressive dysfunction of the day vision or photopic (cone) system with preservation of night vision or scotopic (rod) function. The disease presents with a triad of photophobia, loss of color vision and reduced central vision. This phenotype is distinct from retinitis pigmentosa (RP) in which there are prominent night and peripheral vision disturbances. X-linked cone dystrophy is a genetically heterogeneous disorder, with linkage to loci on Xp11.4--Xp21.1 (COD1, OMIM 304020) and Xq27 (COD2, OMIM 303800). COD1 maps to a region that harbors the RPGR gene, mutations in which account for >70% of patients with X linked RP. The majority of these mutations reside in one purine-rich exon, ORF15, encoding 567 amino acids with a repetitive domain rich in glutamic acid residues. We mapped two families with X-linked cone dystrophy to the COD1 locus and identified two distinct mutations in ORF15 in the RPGR gene (ORF15+1343_1344delGG and ORF15+694_708del15) leading to a frame-shift and premature termination of translation in one case and a deletion of five amino acids in another. Consistent with expression of RPGR in rods and cones, our results show that mutations in RPGR, in addition to X-linked RP, can also cause cone-specific degeneration. PMID- 11875056 TI - Solution structure of human GABA(A) receptor-associated protein GABARAP: implications for biolgoical funcrion and its regulation. AB - Control of neurotransmitter receptor expression and delivery to the postsynaptic membrane is of critical importance for neural signal transduction at synapses. The gamma-aminobutyric acid, type A (GABA(A)) receptor-associated protein GABARAP was reported to have an important role for movement and sorting of GABA(A) receptor molecules to the postsynaptic membrane. GABARAP not only binds to GABA(A) receptor gamma2-subunit but also to tubulin, gephyrin, and ULK1. We present for the first time the high resolution structure of human GABARAP determined by nuclear magnetic resonance in aqueous solution. One part of the molecule, despite being well ordered and rigid on a MHz time scale, exists in at least two different conformations that interchange with each other on a time scale slower than 25 Hz. An important feature of the solution structure is the observation that amino- and carboxyl-terminal ends of the protein directly interact with each other, which is not seen in recently reported crystal structures. The possible biological relevance of these observations for the regulation of GABARAP interactions and functions is discussed. PMID- 11875057 TI - ATM mediates phosphorylation at multiple p53 sites, including Ser(46), in response to ionizing radiation. AB - The p53 tumor suppressor protein preserves genome integrity by regulating growth arrest and apoptosis in response to DNA damage. In response to ionizing radiation (IR), ATM, the gene product mutated in ataxia telangiectasia, stabilizes and activates p53 through phosphorylation of Ser(15) and (indirectly) Ser(20). Here we show that phosphorylation of p53 on Ser(46), a residue important for p53 apoptotic activity, as well as on Ser(9), in response to IR also is dependent on the ATM protein kinase. IR-induced phosphorylation at Ser(46) was inhibited by wortmannin, a phosphatidylinositol 3-kinase inhibitor, but not PD169316, a p38 MAPK inhibitor. p53 C-terminal acetylation at Lys(320) and Lys(382), which may stabilize p53 and activate sequence-specific DNA binding, required Ser(15) phosphorylation by ATM and was enhanced by phosphorylation at nearby residues including Ser(6), Ser(9), and Thr(18). These observations, together with the proposed role of Ser(46) phosphorylation in mediating apoptosis, suggest that ATM is involved in the initiation of p53-dependent apoptosis after IR in human lymphoblastoid cells. PMID- 11875058 TI - The utrophin gene is transcriptionally up-regulated in regenerating muscle. AB - The utrophin gene codes for a large cytoskeletal protein closely related to dystrophin, the gene mutated in Duchenne's muscular dystrophy. Although utrophin could functionally substitute for dystrophin, in Duchenne's muscular dystrophy patients it did not compensate for the absence of dystrophin because in adult muscle utrophin was poorly expressed and limited to subsynaptic nuclei. However, increased levels of utrophin have been observed in regenerated muscles fibers suggesting that utrophin up-regulation in muscle is feasible. We observed that utrophin mRNA was transiently up-regulated at early time points after muscle injury with a peak already 24 h after muscle damage and utrophin induction in activated satellite cells before fusion into young regenerated fibers. Injection of utrophin lacZ constructs into regenerating muscle demonstrated that the utrophin upstream promoter under the control of its intronic enhancer activated the transcription that leads to the expression of the reporter gene in the newly formed fibers, which was not limited to neuromuscular junctions. Utrophin enhancer activity was dependent on an AP-1 site, and in satellite cells of regenerating muscle the AP-1 factors Fra1, Fra2, and JunD were strongly induced. These results establish that utrophin was induced in adult muscle independently from neuromuscular junctions and suggest that growth factors and cytokines that mediate the muscle repair up-regulate utrophin transcription. PMID- 11875059 TI - Pre-existing distortions in nucleic acid structure aid polypurine tract selection by HIV-1 reverse transcriptase. AB - Precise cleavage at the polypurine tract (PPT)/U3 junction by human immunodeficiency virus type 1 (HIV-1) reverse transcriptase RNase H is critical for generating a correct viral DNA end for subsequent integration. Using potassium permanganate (KMnO(4)) modification, we have identified a significant distortion in the nucleic acid structure at the HIV-1 PPT/U3 junction in the absence of trans-acting factors. Unusually high reactivity of template thymine +1 is detected when the PPT primer is extended by DNA or RNA at its 3' terminus. Chemical footprinting suggests that the extent of base unstacking in the wild type species is comparable when the +1 A:T base pair is replaced by a C:T mismatch. However, reactivity of this template base is diminished after alterations to upstream (rA)(4):(dT)(4) or (rG)(6):(dC)(6) tracts. Importantly, there is a correlation between the structural deformation at base pair +1 and precise cleavage at the PPT/U3 junction by HIV-1 reverse transcriptase/RNase H. KMnO(4) modification also revealed unusually high reactivity for one of two (dT)(4):(rA)(4) duplexes upstream of the PPT/U3 junction, suggesting a significant structural distortion within the PPT itself in the absence of the retroviral polymerase. Structural abnormalities in this region are not only essential for resistance of the PPT to hydrolysis but also significantly impact the conformation of the PPT/U3 junction. Our data collectively suggest that the entire PPT sequence contributes to the structural distortion at the PPT/U3 junction, potentially providing a mechanism for its selective processing. PMID- 11875060 TI - Role of sterol regulatory element-binding protein 1 in regulation of renal lipid metabolism and glomerulosclerosis in diabetes mellitus. AB - Diabetic renal disease is associated with lipid deposits in the kidney. The purpose of our study was to determine whether there is altered regulation of the sterol regulatory element-binding proteins (SREBPs) in the diabetic kidney and whether SREBPs mediate the abnormal renal lipid metabolism and diabetic renal disease. In streptozotocin-induced diabetes in the rat, there were marked increases in SREBP-1 and fatty acid synthase (FAS) expression, resulting in increased triglyceride (TG) accumulation. Treatment of diabetic rats with insulin prevented the increased renal expression of SREBP-1 and the accumulation of TG. The role of hyperglycemia in the up-regulation of SREBP-1 was confirmed in renal cells cultured in a high glucose media. High glucose induced increased expression of SREBP-1a and -1c mRNA, SREBP-1 protein, and FAS, resulting in increased TG content. To determine a direct role for SREBP in mediating the increase in renal lipids and glomerulosclerosis, we studied SREBP-1a transgenic mice with increased renal expression of SREBP-1. The increase in SREBP-1 was associated with increased expression of FAS and acetyl CoA carboxylase, resulting in increased TG content, increased expression of transforming growth factor beta1 and vascular endothelial growth factor, mesangial expansion, glomerulosclerosis, and proteinuria. Our study therefore indicates that renal SREBP-1 expression is increased in diabetes and that SREBP-1 plays an important role in the increased lipid synthesis, TG accumulation, mesangial expansion, glomerulosclerosis, and proteinuria by increasing the expression of transforming growth factor beta and vascular endothelial growth factor. PMID- 11875061 TI - Foxa2 (HNF3beta ) controls multiple genes implicated in metabolism-secretion coupling of glucose-induced insulin release. AB - The transcription factor Foxa2 is implicated in blood glucose homeostasis. Conditional expression of Foxa2 or its dominant-negative mutant DN-Foxa2 in INS-1 cells reveals that Foxa2 regulates the expression of genes important for glucose sensing in pancreatic beta-cells. Overexpression of Foxa2 results in blunted glucose-stimulated insulin secretion, whereas induction of DN-Foxa2 causes a left shift of glucose-induced insulin release. The mRNA levels of GLUT2 and glucokinase are drastically decreased after induction of Foxa2. In contrast, loss of Foxa2 function leads to up-regulation of hexokinase (HK) I and II and glucokinase (HK-IV) mRNA expression. The glucokinase and the low K(m) hexokinase activities as well as glycolysis are increased proportionally. In addition, induction of DN-Foxa2 also reduces the expression of beta-cell K(ATP) channel subunits Sur1 and Kir6.2 by 70%. Furthermore, in contrast to previous reports, induction of Foxa2 causes pronounced decreases in the HNF4alpha and HNF1alpha mRNA levels. Foxa2 fails to regulate the expression of Pdx1 transcripts. The expression of insulin and islet amyloid polypeptide is markedly suppressed after induction of Foxa2, while the glucagon mRNA levels are significantly increased. Conversely, Foxa2 is required for glucagon expression in these INS-1-derived cells. These results suggest that Foxa2 is a vital transcription factor evolved to control the expression of genes essential for maintaining beta-cell glucose sensing and glucose homeostasis. PMID- 11875062 TI - Lipoplex-mediated transfection of mammalian cells occurs through the cholesterol dependent clathrin-mediated pathway of endocytosis. AB - Synthetic amphiphiles are widely used as a carrier system. However, to match transfection efficiencies as obtained for viral vectors, further insight is required into the properties of lipoplexes that dictate transfection efficiency, including the mechanism of delivery. Although endocytosis is often referred to as the pathway of lipoplex entry and transfection, its precise nature has been poorly defined. Here, we demonstrate that lipoplex-mediated transfection is inhibited by more than 80%, when plasma membrane cholesterol is depleted with methyl-beta-cyclodextrin. Cholesterol replenishment restores the transfection capacity. Investigation of the cellular distribution of lipoplexes after cholesterol depletion revealed an exclusive inhibition of internalization, whereas cell-association remained unaffected. These data strongly support the notion that complex internalization, rather than the direct translocation of plasmid across the plasma membrane, is a prerequisite for accomplishing effective lipoplex-mediated transfection. We demonstrate that internalized lipoplexes colocalize with transferrin in early endocytic compartments and that lipoplex internalization is inhibited in potassium-depleted cells and in cells overexpressing dominant negative Eps15 mutants. In conjunction with the notion that caveolae-mediated internalization can be excluded, we conclude that efficient lipoplex-mediated transfection requires complex internalization via the cholesterol-dependent clathrin-mediated pathway of endocytosis. PMID- 11875064 TI - Characterizing the DNA contacts and cooperative binding of F plasmid TraM to its cognate sites at oriT. AB - TraM is a DNA binding protein required for conjugative transfer of the self transmissible IncF group of plasmids, including F, R1, and R100. F TraM binds to three sites in F oriT: two high affinity binding sites, sbmA and sbmB, which are direct repeats of nearly identical sequence involved in the autoregulation of the traM gene; and a lower affinity site, sbmC, an inverted repeat important for transfer, which is situated nearest to the nic site where transfer originates. TraM bound cooperatively to its binding sites at oriT; the presence of sbmA and sbmB increased the affinity for sbmC 10-fold. Bending of oriT DNA by TraM was minimal, suggesting that TraM, a tetramer, was able to loop the DNA when bound to sbmA and sbmB simultaneously. Hydroxyl radical footprinting of DNA of sbmA and sbmC revealed that TraM contacted the DNA within a region previously delineated by DNase I footprinting. TraM protected the CT bases within the sequence CTAG, which occurred at 12-base intervals on the top and bottom strand of sbmA, most consistently with other protected bases. The footprint on sbmC revealed that the predicted inverted repeats were protected by TraM with a pattern that began at the center of the repeats and radiated outward at 11-12 base intervals toward the 5'-ends of either strand. At high protein concentrations, this pattern extended beyond the footprint defined by DNase I, suggesting that the DNA was wrapped around the protein forming a nucleosome-like structure, which could aid in preparing the DNA for transfer. PMID- 11875063 TI - The circadian regulatory proteins BMAL1 and cryptochromes are substrates of casein kinase Iepsilon. AB - The serine/threonine protein kinase casein kinase I epsilon (CKIepsilon) is a key regulator of metazoan circadian rhythm. Genetic and biochemical data suggest that CKIepsilon binds to and phosphorylates the PERIOD proteins. However, the PERIOD proteins interact with a variety of circadian regulators, suggesting the possibility that CKIepsilon may interact with and phosphorylate additional clock components as well. We find that CRY1 and BMAL1 are phosphoproteins in cultured cells. Mammalian PERIOD proteins act as a scaffold with distinct domains that simultaneously bind CKIepsilon and mCRY1 and mCRY2 (mCRY). mCRY is phosphorylated by CKIepsilon only when both proteins are bound to mammalian PERIOD proteins. BMAL1 is also a substrate for CKIepsilon in vitro, and CKIepsilon kinase activity positively regulates BMAL1-dependent transcription from circadian promoters in reporter assays. We conclude that CKIepsilon phosphorylates multiple circadian substrates and may exert its effects on circadian rhythm in part by a direct effect on BMAL1-dependent transcription. PMID- 11875065 TI - The yeast glutaredoxins are active as glutathione peroxidases. AB - The yeast Saccharomyces cerevisiae contains two glutaredoxins, encoded by GRX1 and GRX2, which are active as glutathione-dependent oxidoreductases. Our studies show that changes in the levels of glutaredoxins affect the resistance of yeast cells to oxidative stress induced by hydroperoxides. Elevating the gene dosage of GRX1 or GRX2 increases resistance to hydroperoxides including hydrogen peroxide, tert-butyl hydroperoxide and cumene hydroperoxide. The glutaredoxin-mediated resistance to hydroperoxides is dependent on the presence of an intact glutathione system, but does not require the activity of phospholipid hydroperoxide glutathione peroxidases (GPX1-3). Rather, the mechanism appears to be mediated via glutathione conjugation and removal from the cell because it is absent in strains lacking glutathione-S-transferases (GTT1, GTT2) or the GS-X pump (YCF1). We show that the yeast glutaredoxins can directly reduce hydroperoxides in a catalytic manner, using reducing power provided by NADPH, GSH, and glutathione reductase. With cumene hydroperoxide, high pressure liquid chromatography analysis confirmed the formation of the corresponding cumyl alcohol. We propose a model in which the glutathione peroxidase activity of glutaredoxins converts hydroperoxides to their corresponding alcohols; these can then be conjugated to GSH by glutathione-S-transferases and transported into the vacuole by Ycf1. PMID- 11875066 TI - Role of insulin receptor dimerization domains in ligand binding, cooperativity, and modulation by anti-receptor antibodies. AB - To define the structures within the insulin receptor (IR) that are required for high affinity ligand binding, we have used IR fragments consisting of four amino terminal domains (L1, cysteine-rich, L2, first fibronectin type III domain) fused to sequences encoded by exon 10 (including the carboxyl terminus of the alpha subunit). The fragments contained one or both cysteine residues (amino acids 524 and 682) that form disulfides between alpha-subunits in native IR. A dimeric fragment designated IR593.CT (amino acids 1-593 and 704-719) bound (125)I-insulin with high affinity comparable to detergent-solubilized wild type IR and mIR.Fn0/Ex10 (amino acids 1-601 and 650-719) and greater than that of dimeric mIR.Fn0 (amino acids 1-601 and 704-719) and monomeric IR473.CT (amino acids 1-473 and 704-719). However, neither IR593.CT nor mIR.Fn0 exhibited negative cooperativity (a feature characteristic of the native insulin receptor and mIR.Fn0/Ex10), as shown by failure of unlabeled insulin to accelerate dissociation of bound (125)I-insulin. Anti-receptor monoclonal antibodies that recognize epitopes in the first fibronectin type III domain (amino acids 471-593) and inhibit insulin binding to wild type IR inhibited insulin binding to mIR.Fn0/Ex10 but not IR593.CT or mIR.Fn0. We conclude the following: 1) precise positioning of the carboxyl-terminal sequence can be a critical determinant of binding affinity; 2) dimerization via the first fibronectin domain alone can contribute to high affinity ligand binding; and 3) the second dimerization domain encoded by exon 10 is required for ligand cooperativity and modulation by antibodies. PMID- 11875067 TI - Rho activity can alter the translation of p27 mRNA and is important for RasV12 induced transformation in a manner dependent on p27 status. AB - The amount of p27(Kip1) establishes a threshold to which G(1) cyclin-cyclin dependent kinase complexes must surpass prior to cells progressing into S-phase. The amount of p27 is greatest in G(0) cells, intermediate in G(1) cells, and lowest in S-phase cells. However, there is little known regarding the pathways and mechanisms controlling p27 accumulation in G(0) cells. We report that inhibition of Rho, by either lovastatin or C3 exoenzyme, can increase the translational efficiency of p27 mRNA. Similar pharmacologic inhibition of the phosphatidylinositol 3-kinase, the S6 kinase, and the Mek1 kinase pathways all fail to increase translational efficiency in MDA468 cells. This Rho-responsive element lies within a 300-nucleotide region at the 3'-end of the mRNA. By supporting the significance of this signaling pathway to Rho function, we showed that the suppression of Ras(V12) transformation by RhoA(N19) is blocked in p27-/- cells. In contrast this activity is not blocked in Rb-/- or p16-/- cells. The resistance of p27-/- cells to RhoA(N19) is not associated with a failure of RhoA(N19) to accumulate to amounts sufficient to block Rho activity as measured by the organization of actin stress fibers. Together these results indicate a link between Rho and p27. PMID- 11875068 TI - Myogenic differentiation is dependent on both the kinase function and the N terminal sequence of mammalian target of rapamycin. AB - The mammalian target of rapamycin (mTOR) is a serine/threonine protein kinase known to control initiation of translation through two downstream pathways: eukaryotic initiation factor 4E-binding protein 1 (4E-BP1)/eukaryotic initiation factor 4E and ribosomal p70 S6 kinase (S6K1). We previously showed in C2C12 murine myoblasts that rapamycin arrests cells in G(1) phase and completely inhibits terminal myogenesis. To elucidate the pathways that regulate myogenesis, we established stable C2C12 cell lines that express rapamycin-resistant mTOR mutants (mTORrr; S2035I) that have N-terminal deletions (Delta10 or Delta91) or are full-length kinase-dead mTORrr proteins. Additional clones expressing a constitutively active S6K1 were also studied. Our results show that Delta10mTORrr signals 4E-BP1 and permits rapamycin-treated myoblasts to differentiate, confirming the mTOR dependence of the inhibition of myogenesis by rapamycin. C2C12 cells expressing either Delta91mTORrr or kinase-dead mTORrr(D2338A) could not phosphorylate 4E-BP1 in the presence of rapamycin and could not abrogate the inhibition of myogenesis. Taken together, our results indicate that both the kinase function of mTOR and the N terminus (residues 11-91, containing part of the first HEAT domain) are essential for myogenic differentiation. In contrast, constitutive activation of S6K1 does not abrogate rapamycin inhibition of either proliferation or myogenic differentiation. PMID- 11875069 TI - Purification and characterization of a doxorubicin-inhibited NADH-quinone (NADH ferricyanide) reductase from rat liver plasma membranes. AB - Plasma membrane-associated redox systems play important roles in regulation of cell growth, internal pH, signal transduction, apoptosis, and defense against pathogens. Stimulation of cell growth and stimulation of the redox system of plasma membranes are correlated. When cell growth is inhibited by antitumor agents such as doxorubicin, capsaicin, and antitumor sulfonylureas, redox activities of the plasma membrane also are inhibited. A doxorubicin-inhibited NADH-quinone reductase was characterized and purified from plasma membranes of rat liver. First, an NADH-cytochrome b(5) reductase, which was doxorubicin insensitive, was removed from the plasma membranes by the lysosomal protease, cathepsin D. After removal of the NADH-cytochrome b(5) reductase, the plasma membranes retained a doxorubicin-inhibited NADH-quinone reductase activity. The enzyme, with an apparent molecular mass of 57 kDa, was purified 200-fold over the cathepsin D-treated plasma membranes. The purified enzyme had also an NADH coenzyme Q(0) reductase (NADH: external acceptor (quinone) reductase; EC 1.6.5.) activity. Partial amino acid sequence of the enzyme showed that it was unique with no sequence homology to any known protein. Antibody against the enzyme (peptide sequence) was produced and affinity-purified. The purified antibody immunoprecipitated both the NADH-ferricyanide reductase activity and NADH coenzyme Q(0) reductase activity of plasma membranes and cross-reacted with human chronic myelogenous leukemia K562 cells and doxorubicin-resistant human chronic myelogenous leukemia K562R cells. Localization by fluorescence microscopy showed that the reaction was with the external surface of the plasma membranes. The doxorubicin-inhibited NADH-quinone reductase may provide a target for the anthracycline antitumor agents and a candidate ferricyanide reductase for plasma membrane electron transport. PMID- 11875070 TI - ERK8, a new member of the mitogen-activated protein kinase family. AB - The ERKs are a subfamily of the MAPKs that have been implicated in cell growth and differentiation. By using the rat ERK7 cDNA to screen a human multiple tissue cDNA library, we identified a new member of the ERK family, ERK8, that shares 69% amino acid sequence identity with ERK7. Northern analysis demonstrates that ERK8 is present in a number of tissues with maximal expression in the lung and kidney. Fluorescence in situ hybridization localized the ERK8 gene to chromosome 8, band q24.3. Expression of ERK8 in COS cells and bacteria indicates that, in contrast to constitutively active ERK7, ERK8 has minimal basal kinase activity and a unique substrate profile. ERK8, which contains two SH3-binding motifs in its C terminal region, associates with the c-Src SH3 domain in vitro and co immunoprecipitates with c-Src in vivo. Co-transfection with either v-Src or a constitutively active c-Src increases ERK8 activation indicating that ERK8 can be activated downstream of c-Src. ERK8 is also activated following serum stimulation, and the extent of this activation is reduced by pretreatment with the specific Src family inhibitor PP2. The ERK8 activation by serum or Src was not affected by the MEK inhibitor U0126 indicating that activation of ERK8 does not require MEK1, MEK2, or MEK5. Although most closely related to ERK7, the relatively low sequence identity, minimal basal activity, and different substrate profile identify ERK8 as a distinct member of the MAPK family that is activated by an Src-dependent signaling pathway. PMID- 11875071 TI - The peptide repertoires of HLA-B27 subtypes differentially associated to spondyloarthropathy (B*2704 and B*2706) differ by specific changes at three anchor positions. AB - HLA-B*2704 is strongly associated with ankylosing spondylitis. B*2706, which differs from B*2704 by two amino acid changes, is not associated with this disease. A systematic comparison of the B*2704- and B*2706-bound peptide repertoires was carried out to elucidate their overlap and differential features and to correlate them with disease susceptibility. Both subtypes shared about 90% of their peptide repertoires, consisting of peptides with Arg(2) and C-terminal aliphatic or Phe residues. B*2706 polymorphism influenced specificity at three anchor positions: it favored basic residues at P3 and POmega-2 and impaired binding of Tyr and Arg at POmega. Thus, the main structural feature of peptides differentially bound to B*2704 was the presence of C-terminal Tyr or Arg, together with a strong preference for aliphatic/aromatic P3 residues. This is the only known feature of B*2704 and B*2706 that correlates to their differential association with spondyloarthropathy. The concomitant presence of basic P3 and POmega-2 residues was observed only among peptides differentially bound to B*2706, suggesting that it impairs binding to B*2704. Similarity between peptide overlap and the degree of cross-reaction with alloreactive T lymphocytes suggested that the majority of shared ligands maintain unaltered antigenic features in the context of both subtypes. PMID- 11875072 TI - Peroxisome proliferator-activated receptor (PPAR) gamma coactivator-1 recruitment regulates PPAR subtype specificity. AB - The peroxisome proliferator-activated receptors (PPAR) alpha and gamma play key roles in the transcriptional control of contrasting metabolic pathways such as adipogenesis and fatty acid beta-oxidation. Both ligand-activated nuclear receptors bind to common target gene response elements and interact with distinct domains of the transcriptional coactivator PGC-1 to attain their full transcriptional potency. Thus, PPAR subtype specificity may be determined by ligand availability and transcription factor or coactivator expression levels. To identify other, perhaps more precise mechanisms contributing to PPAR subtype specificity, we studied PGC-1 recruitment by PPARs using a previously described hormone response element in the human UCP1 promoter and a human brown adipocyte cell line as our model system. As in rodents, PGC-1 is involved in the transcriptional regulation of the UCP1 gene in humans and mediates the effects of PPARalpha and PPARgamma agonists and retinoic acid. Interestingly, a previously postulated PGC-1 repressor selectively affects the PPARalpha-mediated activation of UCP1 gene expression. Furthermore, inhibition of p38 MAPK signaling, known to regulate the PGC-1/repressor interaction, decreases the stimulatory effect of PPARalpha agonist treatment without reducing the response to thiazolidinedione or retinoic acid. These data support a model whereby PPAR subtype specificity is regulated by recruitment of PGC-1. PMID- 11875073 TI - Fast biphasic regulation of type 3 inositol trisphosphate receptors by cytosolic calcium. AB - In cytosol-like medium (CLM) with a free [Ca(2+)] of 200 nm, a supramaximal concentration of inositol 1,4,5-trisphosphate (IP(3)) (30 microm) evoked (45)Ca(2+) release from type 3 IP(3) receptors only after a latency of 48 +/- 6 ms; this latency could not be reduced by increasing the IP(3) concentration. In CLM containing a low free [Ca(2+)] ( approximately 4 nm), 300 microm IP(3) evoked (45)Ca(2+) release after a latency of 66 +/- 11 ms; this was reduced to 14 +/- 3 ms when the [Ca(2+)] was 1 mm. Preincubation with CLM containing 100 microm Ca(2+) caused a rapid (half-time = 33 +/- 9 ms), complete, and fully reversible inhibition that could not be overcome by a high concentration of IP(3) (300 microm). Hepatic (type 2) IP(3) receptors were not inhibited by Ca(2+) once they had bound IP(3), but 100 microm Ca(2+) rapidly inhibited type 3 IP(3) receptors whether it was delivered before addition of IP(3) or at any stage during a response to IP(3). Ca(2+) increases the affinity of IP(3) for hepatic receptors by slowing IP(3) dissociation, but Ca(2+) had no effect on IP(3) binding to type 3 receptors. The rate of inhibition of type 3 IP(3) receptors by Ca(2+) was faster than the rate of IP(3) dissociation, and occurred at similar rates whether receptors had bound a high (adenophostin) or low affinity (3-deoxy-3-fluoro IP(3)) agonist. Dissociation of agonist is not therefore required for Ca(2+) to inhibit type 3 IP(3) receptors. We conclude that type 2 and 3 IP(3) receptors are each biphasically regulated by Ca(2+), but by different mechanisms. For both, IP(3) binding causes a stimulatory Ca(2+)-binding site to be exposed allowing Ca(2+) to bind and open the channel. IP(3) binding protects type 2 receptors from Ca(2+) inhibition, but type 3 receptors are inhibited by Ca(2+) whether or not they have IP(3) bound. Increases in cytosolic [Ca(2+)] will immediately inhibit type 3 receptors, but inhibit type 2 receptors only after IP(3) has dissociated. PMID- 11875074 TI - Structural characterization of the M* partly folded intermediate of wild type and P138A aspartate aminotransferase from Escherichia coli. AB - A combination of spectroscopic techniques, hydrogen/deuterium exchange, and limited proteolysis experiments coupled to mass spectrometry analysis was used to depict the topology of the monomeric M* partly folded intermediate of aspartate aminotransferase from Escherichia coli in wild type (WT) as well as in a mutant form in which the highly conserved cis-proline at position 138 was replaced by a trans-alanine (P138A). Fluorescence analysis indicates that, although M* is an off-pathway intermediate in the folding of WT aspartate aminotransferase from E. coli, it seems to coincide with an on-pathway folding intermediate for the P138A mutant. Spectroscopic data, hydrogen/deuterium exchange, and limited proteolysis experiments demonstrated the occurrence of conformational differences between the two M* intermediates, with P138A-M* being conceivably more compact than WT-M*. Limited proteolysis data suggested that these conformational differences might be related to a different relative orientation of the small and large domains of the protein induced by the presence of the cis-proline residue at position 138. These differences between the two M* species indicated that in WT-M* Pro138 is in the cis conformation at this stage of the folding process. Moreover, hydrogen/deuterium exchange results showed the occurrence of few differences in the native N(2) forms of WT and P138A, the spectroscopic features and crystallographic structures of which are almost superimposable. PMID- 11875075 TI - Cleavage properties of an archaeal site-specific recombinase, the SSV1 integrase. AB - SSV1 is a virus infecting the extremely thermophilic archaeon Sulfolobus shibatae. The viral-encoded integrase is responsible for site-specific integration of SSV1 into its host genome. The recombinant enzyme was expressed in Escherichia coli, purified to homogeneity, and its biochemical properties investigated in vitro. We show that the SSV1 integrase belongs to the tyrosine recombinases family and that Tyr(314) is involved in the formation of a 3' phosphotyrosine intermediate. The integrase cleaves both strands of a synthetic substrate in a temperature-dependent reaction, the cleavage efficiency increasing with temperature. A discontinuity was observed in the Arrhenius plot above 50 degrees C, suggesting that a conformational transition may occur in the integrase at this temperature. Analysis of cleavage time course suggested that noncovalent binding of the integrase to its substrate is rate-limiting in the cleavage reaction. The cleavage positions were localized on each side of the anticodon loop of the tRNA gene where SSV1 integration takes place. Finally, the SSV1 integrase is able to cut substrates harboring mismatches in the binding site. For the cleavage step, the chemical nature of the base in position -1 of cleavage seems to be more important than its pairing to the opposite strand. PMID- 11875076 TI - Functional characterization of the G protein regulator RGS13. AB - The signaling cascades evoked by G protein-coupled receptors are a predominant mechanism of cellular communication. The regulators of G protein signaling (RGS) comprise a family of proteins that attenuate G protein-mediated signal transduction. Here we report the characterization of RGS13, the smallest member of the RGS family, which has been cloned from human lung. RGS13 has been found most abundantly in human tonsil, followed by thymus, lung, lymph node, and spleen. RGS13 is a GTPase-activating protein for Galpha(i) and Galpha(o) but not Galpha(s). RGS13 binds Galpha(q) in the presence of aluminum magnesium fluoride, suggesting that it bears GTPase-activating protein activity toward Galpha(q). RGS13 blocks MAPK activity induced by Galpha(i)- or Galpha(q)-coupled receptors. RGS13 also attenuates GTPase-deficient Galpha(q) (Galpha(q)QL) mediated cAMP response element activation but not transcription evoked by constitutively active Galpha(12) or Galpha(13). Surprisingly, RGS13 inhibits cAMP generation elicited by stimulation of the beta(2)-adrenergic receptor. These data suggest that RGS13 may regulate Galpha(i)-, Galpha(q)-, and Galpha(s)-coupled signaling cascades. PMID- 11875077 TI - Dominant negative Rab3D inhibits amylase release from mouse pancreatic acini. AB - Rab3 proteins are believed to play an important role in regulated exocytosis and previous work has demonstrated the presence of Rab3D on pancreatic zymogen granules. To further understand the function of Rab3D in acinar cell exocytosis, adenoviral constructs were prepared encoding hemagglutinin-tagged wild type Rab3D and three mutant forms, N135I and T36N (both deficient in guanine nucleotide binding) and Q81L (deficient in GTP hydrolysis), which also expressed enhanced green fluorescent protein driven by a separate promoter. When isolated mouse pancreatic acini were cultured with 5 x 10(6) pfu/ml adenovirus, nearly 100% of acini were infected as visualized by expression of green fluorescent protein. Cultured acini showed a biphasic dose-response to cholecystokinin (CCK); basal amylase secretion was 1.8 +/- 0.3%/30 min, peak release was 7.3 +/- 0.2%/30 min at 30 pm CCK and reduced secretion was observed at higher CCK concentrations. Control beta-galactosidase virus infection had no effect on either basal or CCK induced secretion in the titer range from 0.5 to 10 x 10(6) pfu/ml. While the expression of Rab3D and Rab3D Q81L had no effect on amylase secretion, Rab3D N135I and T36N functioned as dominant negative mutants and inhibited CCK-induced amylase release by 40-50% at all points on the CCK dose-response curve from 3 to 300 pm. Inhibition was stronger during the first 5 min (71 +/- 5%) than over 30 min (36%+/-5%). Similar inhibition was found using other agonists including bombesin, carbachol, and cAMP. Localization of adenoviral expressed Rab protein showed wild type Rab3D localized to zymogen granules. The two dominant negative mutants did not localize to granules and were primarily in the basolateral region of the cell. Since both dominant negative Rab3D mutants had no effect on intracellular calcium increase induced by CCK, it is unlikely that they acted at receptors or transmembrane signaling. These results suggest that Rab3D plays an important role in regulating the terminal steps of acinar exocytosis and that this effect is greatest on the early phase of amylase release. PMID- 11875078 TI - Caspase proteolysis of the cohesin component RAD21 promotes apoptosis. AB - Caspases are a conserved family of proteases that play a critical role in the execution of apoptosis by cleaving key cellular proteins at Asp residues and modifying their function. Using an expression cloning strategy we recently developed, we isolated human RAD21/SCC1/MCD1 as a novel caspase substrate. RAD21 is a component of the cohesin complex that holds sister chromatids together during mitosis and repairs double-strand DNA breaks. Interestingly, RAD21 is cleaved by a caspase-like Esp1/separase at the onset of anaphase to trigger sister chromatid separation. Here, we demonstrate that human RAD21 is preferentially cleaved at Asp(279) by caspases-3 and -7 in vitro to generate two major proteolytic products of approximately 65 and 48 kDa. Moreover, we show that RAD21 is specifically proteolyzed by caspases into a similarly sized 65-kDa carboxyl-terminal product in cells undergoing apoptosis in response to diverse stimuli. We also demonstrate that caspase proteolysis of RAD21 precedes apoptotic chromatin condensation and has important functional consequences, viz. the partial removal of RAD21 from chromatin and the production of a proapoptotic carboxyl-terminal cleavage product that amplifies the cell death signal. Taken together, these findings point to an entirely novel function of RAD21 in the execution of apoptosis. PMID- 11875079 TI - Genetic fusions of globular proteins to the epsilon subunit of the Escherichia coli ATP synthase: Implications for in vivo rotational catalysis and epsilon subunit function. AB - The rotational mechanism of ATP synthase was investigated by fusing three proteins from Escherichia coli, the 12-kDa soluble cytochrome b(562), the 20-kDa flavodoxin, and the 28-kDa flavodoxin reductase, to the C terminus of the epsilon subunit of the enzyme. According to the concept of rotational catalysis, because epsilon is part of the rotor a large domain added at this site should sterically clash with the second stalk, blocking rotation and fully inhibiting the enzyme. E. coli cells expressing the cytochrome b(562) fusion in place of wild-type epsilon grew using acetate as the energy source, indicating their capacity for oxidative phosphorylation. Cells expressing the larger flavodoxin or flavodoxin reductase fusions failed to grow on acetate. Immunoblot analysis showed that the fusion proteins were stable in the cells and that they had no effect on enzyme assembly. These results provide initial evidence supporting rotational catalysis in vivo. In membrane vesicles, the cytochrome b(562) fusion caused an increase in the apparent ATPase activity but a minor decrease in proton pumping. Vesicles bearing ATP synthase containing the larger fusion proteins showed reduced but significant levels of ATPase activity that was sensitive to inhibition by dicyclohexylcarbodiimide (DCCD) but no proton pumping. Thus, all fusions to epsilon generated an uncoupled component of ATPase activity. These results imply that a function of the C terminus of epsilon in F(1)F(0) is to increase the efficiency of the enzyme by specifically preventing the uncoupled hydrolysis of ATP. Given the sensitivity to DCCD, this uncoupled ATP hydrolysis may arise from rotational steps of gammaepsilon in the inappropriate direction after ATP is bound at the catalytic site. It is proposed that the C-terminal domain of epsilon functions to ensure that rotation occurs only in the direction of ATP synthesis when ADP is bound and only in the direction of hydrolysis when ATP is bound. PMID- 11875080 TI - Regulation of erythropoietin-induced STAT serine phosphorylation by distinct mitogen-activated protein kinases. AB - The STAT proteins are a family of latent transcription factors that are activated by a wide variety of cytokines. Upon receptor engagement, STATs become tyrosine phosphorylated, translocate to the nucleus, and induce expression of target genes. In addition to tyrosine phosphorylation, maximal activation of some STAT proteins requires serine phosphorylation within the transactivation domain. Here we focus on STAT phosphorylation after engagement of the erythropoietin receptor (EPO-R). In Ba/F3-EPO-R cells, EPO induces tyrosine and serine phosphorylation of STAT1, STAT3, STAT5A, and STAT5B. Identical regions of the EPO-R couple to both tyrosine and serine phosphorylation of each cognate STAT protein. A proximal region of the EPO-R lacking cytoplasmic tyrosines couples to STAT1 and STAT3 phosphorylation as well as ERK and p38(HOG) activation, but not JNK/SAPK. STAT1 serine phosphorylation was perturbed by inhibition of ERK and p38 pathways, whereas only inhibition of ERK activation blocked STAT3 serine phosphorylation in response to EPO. STAT5A/B phosphorylation is downstream of EPO-R Tyr(343), however, STAT5A/B serine phosphorylation is unaffected by either ERK or p38 inhibition. Physiological responses induced by EPO may depend on regulation of serine phosphorylation of the STAT molecules by p38(HOG) and the ERK family of kinases as well as additional serine/threonine kinases. PMID- 11875081 TI - IRB approval--who needs it? PMID- 11875082 TI - Society of Vascular Surgery arteriovenous access reporting standards. PMID- 11875083 TI - Platelet aggregation inhibitors for use in peripheral vascular interventions: what can we learn from the experience in the coronary arteries? AB - During the last decade, an enormous amount of information has been gathered about the function of the platelet and its impact on percutaneous vascular interventions. With the discovery of the GP IIb/IIIa receptor, which is responsible for platelet aggregation, new drug antagonists have been developed to prevent platelet aggregation that may result in arterial thrombosis or platelet microembolization. These drugs include the three GP IIb/IIIa receptor antagonists approved by the Food and Drug Administration: abciximab (ReoPro), eptifibatide (Integrilin), and tirofiban (Aggrastat). These drugs have been used in several large studies to improve the outcome of coronary interventions and in conjunction with plasminogen activators to accelerate thrombolysis. In addition, because no oral GP IIb/IIIa inhibitor exists, other oral regimens have been developed with use of the thienopyridines, ticlopidine (Ticlid) and clopidogrel (Plavix), in combination with aspirin to prevent platelet aggregation and thrombosis. Because the majority of investigations have been performed in patients undergoing coronary interventions, knowledge of these data is necessary to attempt to translate the use of these antiplatelet drugs to peripheral vascular interventions. The goal of this article is to review the use of these agents in the percutaneous treatment of coronary artery disease and give insight to their potential utility in noncoronary interventions. PMID- 11875084 TI - Research publications in vascular and interventional radiology: research topics, study designs, and statistical methods. AB - PURPOSE: Statistical analysis is the universal language of medical research and is a vital tool for communicating the results of vascular and interventional radiology (VIR) procedures. Major articles in two radiology journals were surveyed to characterize the research topics, study designs, and statistical methods seen in recent VIR research publications. MATERIALS AND METHODS: The authors retrospectively reviewed 130 major clinical VIR articles published from July 2000 to June 2001: 72 articles (55%) from JVIR and 58 articles (45%) from RADIOLOGY: Articles were categorized by research topic and study design. Data were collected on the statistical methodology of each article. RESULTS: Research topics included vascular intervention in 65 of 130 articles (50%), nonvascular intervention in 26 (20%), vascular imaging in 23 (18%), biopsy in nine (7%), and other topics in seven (5%). Study design was descriptive in 87 studies (67%), comparative in 39 studies (30%), and involved secondary data analysis in four studies (3%). Of 126 primary clinical studies, outcome was cross-sectional (assessed at a single time point) in 40 studies (32%) and longitudinal (measured over time) in 86 studies (68%). Median sample size was 61. Basic tests of association (t-test, chi(2) test, etc.) were used in 71 articles (56%) and advanced tests of association (regression analysis) were presented in 25 (20%). Survival analysis was applied in 34 articles (27%). Decision statistics such as sensitivity/specificity were not used commonly (12%). Confidence intervals and power calculations were reported infrequently (15% and 7%, respectively). CONCLUSIONS: VIR publications focus on time-dependent outcomes after therapeutic interventions. Readers should understand basic tests of association and survival analysis--these include only 20 named statistical tests. PMID- 11875085 TI - Endovascular placement of self-expanding nitinol coil stents for the treatment of femoropopliteal obstructive disease. AB - PURPOSE: To report on a prospective two-center study to evaluate safety and effectiveness of a self-expanding nitinol coil stent in patients with femoropopliteal obstructive disease. MATERIALS AND METHODS: The IntraCoil nitinol stent was used in 37 patients (28 men and nine women; mean age, 62.4 y plus minus 10.7; range, 43-81 y) presenting with high-grade stenoses (n = 23) or short (<3 cm) occlusions (n = 17) of the superficial femoral artery (SFA; n = 33) or popliteal artery (n = 4). Indications for stent placement were significant residual stenosis (>30%) or dissection after angioplasty. Follow-up evaluations with measurement of the Doppler ankle-brachial index (ABI), assessment of Rutherford clinical stage, and color-coded duplex sonography were performed at discharge and 1, 3, 6, 12, and 18 months thereafter. Primary endpoints of the study were immediate technical and clinical success and 1-year patency. RESULTS: Initial technical success was achieved in all patients. In 10 patients (27%), more than one 40-mm-long device had to be implanted for total lesion coverage; in three patients (8.1%), stents were placed in two separate segments of the SFA simultaneously. The total number of stents deployed was 50. Stent placement induced an initial improvement of the ABI from 0.54 plus minus 0.2 to 0.92 plus minus 0.11 (P <.01). Follow-up data for 12 months after treatment are available for 29 of 37 patients (78.4%); mean follow-up is 15.6 months (range, 1-26 mo). Primary patency rates at 6 and 12 months were 97.1% (SE = 2.9) and 86.2% (SE = 6.5). The primary assisted patency rate was 100% at 12 months. CONCLUSION: Endovascular placement of the IntraCoil self-expanding nitinol coil stent for salvage of failed angioplasty in patients with femoropopliteal obstructive disease is an effective and safe procedure with promising mid-term results. PMID- 11875086 TI - Main portal vein access in transjugular intrahepatic portosystemic shunt procedures: use of three-dimensional ultrasound to ensure safety. AB - PURPOSE: To document the safety of main portal vein (PV) access to create transjugular portosystemic intrahepatic shunts (TIPS), provided that three dimensional ultrasonography (3D US) can document the puncture to have entered a surface of the PV suitable for tamponade. MATERIALS AND METHODS: In 11 patients who underwent conventional TIPS creation (n = 10) or a transjugular portacaval shunt procedure (n = 1), the puncture was found angiographically to enter the main PV. In seven cases, this occurred at the PV bifurcation and, in four, it occurred in the superior third of the PV. 3D US was used to determine whether the point of PV entry was functionally intrahepatic or extrahepatic. The puncture site was deemed to be intrahepatic if liver covered the puncture site on all three orthogonal imaging planes (sagittal, coronal, and transverse). If the puncture site was surrounded by liver, the access was used to deploy a metallic stent (uncovered, n = 10; covered, n = 1). Medical records and follow-up cross sectional imaging studies were reviewed for evidence of hemorrhage complications. Pathologic correlation was performed in one explanted liver and autopsy specimens in five other patients. RESULTS: In nine of 11 patients, 3D US was diagnostic and confidently verified that liver completely covered the portal vein access site. In two patients with diagnostically uncertain 3D US results, transcatheter injection of contrast medium documented no extravasation. All TIPS and direct portacaval shunt procedures were technically successful. No hemorrhagic complications occurred. Examination of pathologic specimens documented this portion of the portal vein to be extraperitoneal, but attached to the superior surface of the caudate lobe with fibrous tissue and small portal vein branches. CONCLUSIONS: The bifurcation and posterior aspect of the superior third of the main PV can be safely used for TIPS procedures, provided access is proven to be surrounded by liver. 3D US can usually confidently determine if the PV entry site is functionally intrahepatic. PMID- 11875087 TI - Malignant gastric outlet obstructions: treatment by means of coaxial placement of uncovered and covered expandable nitinol stents. AB - PURPOSE: To assess whether coaxial placement of uncovered and covered expandable nitinol stents overcomes the disadvantages of the increased migration rate seen with covered stents and the tumor ingrowth seen in uncovered stents in the treatment of malignant gastric outlet obstructions. MATERIALS AND METHODS: Two types of expandable nitinol stent were designed: an uncovered stent and a covered stent. Under fluoroscopic guidance, the uncovered and covered stents were placed coaxially with complete overlap in 39 consecutive patients with malignant gastric outlet obstruction caused by stomach cancer. Food intake capacity was graded on a scale of 0-4. Stent patency rate was estimated by the Kaplan-Meier method. RESULTS: Technical success rate was 97% (38 of 39 patients). After stent placement, food intake capacity improved at least one grade in 36 patients. Stent migration occurred in three patients (8%), that is, partial (n = 2) or complete (n = 1) upward migration of the inner covered stent into the stomach. Two of these patients were treated by placement of an additional covered stent. During the mean follow-up period of 134 days (range, 15-569 d), 10 patients developed recurrent symptoms of obstruction with tumor overgrowth being the most common cause. Nine underwent placement of an additional covered stent with good results. The median period of primary stent patency was 157 days (mean, 278 d). The 30-, 60-, and 180-day patency rates were 97%, 91%, and 39%, respectively. Four patients (10%) died within 1 month after the procedure. CONCLUSION: Coaxial stent placement technique seems to contribute to decreasing the migration rate of the stent and decrease the rate of recurrent obstruction by preventing or delaying tumor ingrowth. PMID- 11875088 TI - Retrievable covered nitinol stents: experiences in 108 patients with malignant esophageal strictures. AB - PURPOSE: The authors report their experience with three types of retrievable covered nitinol stents in patients with malignant esophageal strictures. MATERIALS AND METHODS: Three types of retrievable covered nitinol stents were designed. Type A stents were placed in 45 patients, type B stents were placed in 29 patients, and type C stents were placed in 34 patients. The stents were removed with use of a stent retrieval set under fluoroscopic guidance when the stents caused complications. Stent patency, symptom relief, survival rate, and complications were analyzed relative to stent type and radiation therapy. RESULTS: The timing of radiation and the stent type have significant effects on occurrence of complications such as stent migration and fistula formation (P =.002 and P = 0.029, respectively). Complications were significantly more frequent in patients with the type B stent than those with type A or type C stents (P =.008). Patients who underwent radiation therapy before stent placement or who underwent no radiation therapy experienced substantially less complications than those who underwent radiation therapy after stent placement (P =.005 and P <.001, respectively). The survival period was significantly longer in patients who underwent radiation therapy after stent placement than in the other groups (P =.034). Stents were removed from 15 patients (14%) 2 days to 16 weeks (mean, 4 weeks) after stent placement as a result of severe pain (n = 7), stent migration (n = 6), or stent deformity (n = 2). Stent removal was well tolerated in all patients. CONCLUSION: Use of retrievable covered nitinol stents seems to be a safe and effective method of treatment in patients with malignant esophageal strictures. However, removal of the stents was needed in 14% of the patients because of complications. Patients who underwent radiation therapy after stent placement and those with the type B stent experienced more complications than other patients. PMID- 11875089 TI - Nonsurgically placed nasolacrimal stents for epiphora: long-term results and factors favoring stent patency. AB - PURPOSE: To evaluate long-term effectiveness of a polyurethane stent in lacrimal system treatment. MATERIALS AND METHODS: Fluoroscopically guided placement of a polyurethane lacrimal stent was evaluated in 727 obstructed lacrimal systems in 588 patients with a mean follow-up period of 77 weeks (range, 1-277 wk), from which the technical and initial success rates and long-term patency were calculated. The impact of factors affecting long-term patency, including symptom duration, previous probing, presence of pus or eye discharge, lesion multiplicity, and site of obstruction were evaluated. Changes in 295 patients' lacrimal sac configuration after stent removal were analyzed and classified into three categories: contraction, widening, and no change. RESULTS: The overall technical success rate in this study was 95.9%, with a 90.8% initial clinical success rate. The median primary patency duration was 504.33 days plus minus 23.17; secondary median patency duration was 642.87 days plus minus 24.68. Most favorable factors such as symptom duration, previous probing, presence of pus, or eye discharge showed no significant difference; however, site of obstruction and lesion multiplicity influenced mean patency. Lesions below the junction and single lesions showed longer patency periods. Common canalicular obstruction after stent removal occurred in 50 (17%) lacrimal systems. Irregular sac configuration appeared in 171 (58%) lacrimal systems after stent removal. Sac configuration was the same in 185 (62.7%) lacrimal systems, contracted in 105 (35.6%), and widened in five (1.7%). In 151 eyes of 295, epiphora recurred 121.6 days (range, 1-1,182 d) after stent removal. CONCLUSION: Fluoroscopic lacrimal stent placement is a simple and safe outpatient procedure, but high recurrence and sac irregularities after stent removal are not encouraging. PMID- 11875090 TI - Experimental evaluation of a new transcatheter vascular embolization device in the swine model. AB - PURPOSE: To evaluate a new transcatheter device suitable for arterial embolization in an animal model. MATERIALS AND METHODS: A new prototype self expanding braided embolic device (Embolizor), consisting of nitinol wire strands fixed at either end with platinum-iridium bands and covered with a film of polyethylene, was deployed through 5-F diagnostic catheters into renal artery branches in five swine. Standard stainless-steel spring coils were deployed in other renal branches and served as controls. The animals underwent follow-up selective renal arteriography to determine presence or absence of vessel recanalization or device migration 15-23 days after device deployment. Histopathologic evaluation of target vessels and peripheral renal parenchyma was also performed. RESULTS: Ten Embolizors and 10 stainless-steel coils were deployed in arteries ranging in size from 1.8 to 3.0 mm in diameter. The Embolizor was easily and precisely deployed. Angiographic evidence of vascular occlusion in the Embolizor group was noted within 30 seconds in eight device deployments and within 5 minutes in two. No early or delayed device migration was noted on follow-up arteriography. In the control group, seven of 10 previously occluded arteries were recanalized. No recanalization was noted in the Embolizor group. Light microscopy revealed evidence of infarction in all specimens examined. Whereas three specimens in the Embolizor group contained occasional giant cells, there were numerous multinucleated giant cells present within the interstices of all control spring coils. CONCLUSION: The Embolizor was easily, precisely, and successfully deployed through standard selective diagnostic angiographic catheters. Short-term follow-up demonstrated no recanalization or migration of the device. The Embolizor was shown on histopathologic analysis to have no significant foreign body reaction. PMID- 11875091 TI - Percutaneous radiofrequency ablation therapy after intrathoracic saline solution infusion for liver tumor in the hepatic dome. AB - Two liver tumors undetected by ultrasonography (US) because they were located in the hepatic dome were treated with radiofrequency (RF) ablation therapy after intrathoracic saline solution infusion. After administration of local anesthesia, artificial pneumothorax was produced by needle thoracentesis and a drainage catheter was inserted into the right thoracic cavity. After saline solution (450 500 mL) was injected into the thoracic cavity via the catheter, US-guided RF ablation was performed. No severe complications occurred and complete therapeutic effects were obtained. Percutaneous RF ablation therapy with intrathoracic saline solution injection seems to be a feasible alternative to other ablation therapies. PMID- 11875092 TI - Radiofrequency ablation for hemobilia secondary to hepatocellular carcinoma. AB - Hemobilia is a rare manifestation of hepatic malignancies. The current treatment of choice for hemobilia is transcatheter hepatic arterial embolization. However, there have been only two published reports that describe the use of hepatic arterial embolization for hemobilia caused by hepatic neoplasms. In addition, this procedure is occasionally unsuccessful in the treatment of hemobilia. A case in which hemobilia caused by hepatocellular carcinoma was successfully treated with percutaneous radiofrequency tumor ablation after several failed hepatic arterial embolizations is described in this report. PMID- 11875093 TI - Aneurysm expansion after stent-graft placement in the absence of endoleak. AB - Sixty-three patients with thoracic or abdominal aortic aneurysms were treated with endovascular stent-grafts. No endoleak was identified at any interval of follow-up in 58 patients. In four of them (7%), the aneurysms expanded by 10 mm or more during follow-up and additional interventions were required. Aneurysm expansion was caused by inappropriate sealing at the aneurysmal necks in two patients and transgraft seroma in the other two. Although some aneurysm expansion could be avoided by proper patient selection and accurate placement of stent grafts, it seems difficult to predict aneurysm expansion in most cases. PMID- 11875094 TI - Cutting balloon angioplasty for resistant renal artery in-stent restenosis. AB - A 76-year-old woman presented with recurrent arterial hypertension 6 months after uncomplicated primary renal artery stent placement. Diagnostic arteriography revealed severe renal artery in-stent restenosis. On repeat intervention, the lesion was resistant to attempted conventional percutaneous transluminal angioplasty (PTA) with unchanged systolic pressure gradients across the stent. Cutting balloon angioplasty (CBA) was performed with use of a 4-mm cutting balloon (IVT, San Diego, CA). CBA successfully reduced the pressure gradient to below the level of significance. Subsequent conventional PTA enhanced the lumen diameter inside the stent. The arterial hypertension reverted to normal values and duplex ultrasonography (US) at 10-month follow-up demonstrated normal renal artery hemodynamics without stenosis. CBA for potential use in renal artery in stent restenosis and other peripheral neointimal hyperplasia is discussed. PMID- 11875095 TI - Resolution of secondary pulmonary arteriovenous malformations after embolization of a congenital superior-mesenteric-vein-to-left-renal-vein shunt. AB - A 7-year-old boy who presented with cyanosis and exercise intolerance was diagnosed with pulmonary arteriovenous malformations (PAVMs) secondary to a congenital superior-mesenteric-vein-to-left-renal-vein (SMV-LRV) shunt. Resolution of the PAVMs (confirmed by chest computed tomography [CT]) occurred after embolization of this shunt. Although the pathogenesis of his PAVM is not well known, this case demonstrated a close relationship between a congenital SMV LRV shunt and the development of PAVM. PMID- 11875096 TI - Thoracic duct injury associated with left internal jugular vein catheterization: anatomic considerations. AB - Ultrasound (US)-guided cannulation of the internal jugular vein (IJV) has become the preferred approach for venous access as a result of its higher success rate and lower incidence of complications. This report describes a case of thoracic duct injury during US-guided left IJV catheterization. The normal and variant anatomy of the thoracic duct in the neck is illustrated. PMID- 11875097 TI - Transoral vertebroplasty for a fractured C2 aneurysmal bone cyst. PMID- 11875098 TI - Re: A randomized, prospective comparison of the Tesio, Ash Split, and Opti-flow hemodialysis catheters. PMID- 11875099 TI - GnRH activates ERK1/2 leading to the induction of c-fos and LHbeta protein expression in LbetaT2 cells. AB - GnRH acts on pituitary gonadotropes to stimulate the synthesis and release of LH and FSH. However, the signaling pathways downstream of the GnRH receptor that mediate these effects are not fully understood. In this paper, we demonstrate that GnRH activates ERK, c-Jun N-terminal kinase, and p38MAPK in the LbetaT2 gonadotrope cell line. Phosphorylation of both ERK and p38MAPK are stimulated rapidly, 30- to 50-fold in 5 min, but activation of c-Jun N-terminal kinase has slower kinetics, reaching only 10-fold after 30 min. Activation of ERK by GnRH is blocked by inhibition of MAPK kinase (MEK) and partially blocked by inhibition of PKC and calcium, but not PI3K or p38MAPK signaling. We demonstrate that phosphorylated ERK accumulates in the nucleus in a PKC-dependent manner. We also show that GnRH induces c-fos and LHbeta subunit protein expression in LbetaT2 cells via MEK. Experiments with EGTA or calcium channel antagonists indicated that calcium influx is important for the induction of both genes by GnRH. In conclusion, these results show that GnRH activates all three MAPK subfamilies in LbetaT2 cells and induces c-fos and LHbeta protein expression through calcium and MEK-dependent mechanisms. These results also demonstrate that the nuclear translocation of ERK by GnRH requires PKC signaling. PMID- 11875100 TI - Identification of a discrete promoter region of the human GnRH gene that is sufficient for directing neuron-specific expression: a role for POU homeodomain transcription factors. AB - The human GnRH (hGnRH) gene is expressed, and the GnRH decapeptide produced, primarily in the GnRH neurons of the diencephalon. The molecular elements important for the cell-specific expression and regulation of the hGnRH gene are not well established at this time; therefore, we have used a transgenic mouse model to isolate cis-regulatory elements important for directing gene expression to GnRH neurons in the hypothalamus. Gene constructs consisting of various promoter deletion fragments of the hGnRH gene fused to the luciferase (LUC) reporter gene have been used to create transgenic mouse lines. Cell-specific expression, with the criterion being luciferase expression directed to GnRH neurons of the hypothalamus, was observed when 992 bp, but not 795 bp, of the hGnRH gene promoter were used. Tissue-specific expression was also observed when a deletion construct containing the region from -992 to -763 was fused to a minimal 48-bp promoter fragment fused to LUC. These data indicate that the region between -992 and -795 contains elements both essential and sufficient for targeting gene expression to GnRH neurons. This promoter region was found to contain two DNA-binding sites for the POU class of transcription factors, each of which specifically interacted with the POU homeodomain proteins Brn-2 and Oct-1. Functional studies demonstrated that Brn-2 increased promoter activity of the human and mouse GnRH genes. PMID- 11875102 TI - Decreased expression of the GHRH receptor gene due to a mutation in a Pit-1 binding site. AB - A variety of mutations in the gene encoding the GHRH receptor (GHRHR) that are predicted to alter protein structure or function have been recently described in patients with isolated GH deficiency type IB. In the present report we describe a patient with isolated GH deficiency type IB who was heterozygous for two novel mutations in this gene: a missense mutation in codon 329 that replaces lysine with glutamic acid (K329E) and an A-->C transversion (position -124) in one of the two sites of the promoter region that binds the pituitary-specific transcription factor Pit-1, which is required for GHRHR expression. Chinese hamster ovary cells that were transfected with a cDNA encoding the K329E GHRHR expressed the receptor but failed to show a cAMP response after treatment with GHRH, confirming the lack of functionality. To test the effect of the A-->C mutation at position -124 of the promoter, we transfected rat GH3 pituitary cells, which express endogenous Pit-1, with plasmids in which the luciferase reporter gene was under the control of either the wild-type or the mutant promoter. GH3 cells expressing the mutant promoter showed significantly less luciferase activity than cells expressing the wild-type promoter. DNA-binding studies confirmed that the A-->C base change markedly reduces DNA binding to the Pit-1 protein. These results demonstrate that mutations in the GHRHR are not limited to the coding sequence and that promoter mutations that impair Pit-1 binding can reduce expression of the GHRHR gene. PMID- 11875103 TI - A negative coregulator for the human ER. AB - ERalpha is a ligand-activated transcription factor and a key regulator of the processes involved in cellular proliferation and differentiation. In addition, aberrant ERalpha activity is linked to several pathological conditions including breast cancer. A complex network of coregulatory proteins is largely believed to determine the transcriptional activity of ERalpha. We report here the isolation of a protein, denoted RTA for repressor of tamoxifen transcriptional activity, which contains an RNA recognition motif and interacts with the receptor N terminal activation domain. RTA interacts with RNA in vitro, and its overexpression inhibits the partial agonist activity manifest by the antiestrogen tamoxifen while minimally affecting E2-activated transcription. Mutation of the RNA recognition motif alters RNA binding specificity and results in a dominant negative form of RTA that leads to derepression of ERalpha transcriptional activity, allowing all classes of antiestrogens to manifest partial agonist activity and enhancing agonist efficacy. These findings suggest a role for RNA binding proteins as coregulatory factors of the nuclear receptor family and reveal a novel mechanism by which antiestrogens can manifest agonist activities in some tissues. PMID- 11875105 TI - Allosteric regulation of estrogen receptor structure, function, and coactivator recruitment by different estrogen response elements. AB - Hormone-activated ERs (ERalpha and ERbeta) bind with high affinity to specific DNA sequences, estrogen response elements (EREs), located within the regulatory regions of target genes. Once considered to function solely as receptor tethers, there is an increasing amount of recent evidence to suggest that the sequence of the ERE can influence receptor activity. In this study, we have performed a systematic analysis of the role of different EREs in ER pharmacology. Specifically, by measuring ER activity on the vitellogenin A2, complement 3 gene, pS2, and lactoferrin EREs, we demonstrate that the activities of E2 and xenoestrogen ligands through ERalpha and ERbeta are significantly influenced by the nature of the response element. Using a series of ERalpha and ERbeta interacting peptides that contain the coactivator-binding motif LXXLL, we show that the type of ERE with which the receptor associates regulates the structure of the coactivator pocket on ER. Furthermore, using a novel ELISA developed to measure ER-coactivator interactions revealed that these different conformational states of ERalpha and ERbeta are functionally relevant, as they dictate receptor coactivator binding preference. Together, these results indicate that the DNA response element is a key regulator of receptor structure and biological activity and suggest the ERE sequence influences the recruitment of coactivators to the ER at target gene promoters. We propose that DNA-induced alteration of protein structure and coregulator recruitment may serve as a universal regulatory component for differential gene expression by other nuclear hormone receptors and unrelated transcription factors. PMID- 11875107 TI - Ligand-selective interactions of ER detected in living cells by fluorescence resonance energy transfer. AB - Some aspects of ligand-regulated transcription activation by the estrogen receptor (ER) are associated with the estrogen-dependent formation of a hydrophobic cleft on the receptor surface. At least in vitro, this cleft is required for direct interaction of ER with an alpha helix, containing variants of the sequence LXXLL, found in many coactivators. In cells, it is unknown whether ER interactions with the different LXXLL-containing helices are uniformly similar or whether they vary with LXXLL sequence or activating ligand. Using fluorescence resonance energy transfer (FRET), we confirm in the physiological environment a direct interaction between the estradiol (E2)-bound ER and LXXLL peptides expressed in living cells as fusions with spectral variants of the green fluorescent protein. This interaction was blocked by a single amino acid mutation in the hydrophobic cleft. No FRET was detected when cells were incubated with the antiestrogenic ligands tamoxifen and ICI 182,780. E2, diethylstilbestrol, ethyl indenestrol A, and 6,4'-dihydroxyflavone all promoted FRET and activated ER dependent transcription. Measurement of the level of FRET of ER with different LXXLL-containing peptides suggested that the orientations or affinities of the LXXLL interactions with the hydrophobic cleft were globally similar but slightly different for some activating ligands. PMID- 11875108 TI - A nonclassical estrogen membrane receptor triggers rapid differential actions in the endocrine pancreas. AB - Glucose homeostasis in blood is mainly maintained by insulin released from beta cells and glucagon released from alpha-cells, both integrated within the pancreatic islet of Langerhans. The secretory processes in both types of cells are triggered by a rise in intracellular calcium concentration ([Ca2+](i)). In this study, rapid effects of the natural hormone E2 on [Ca2+](i) were studied in both types of cells within intact islets using laser scanning confocal microscopy. alpha- And beta-cells showed opposite [Ca2+](i) responses when stimulated with physiological concentrations of 17beta-E2. Although the estrogen produced an increase in the frequency of glucose-induced [Ca2+](i) oscillations in insulin-releasing beta-cells, it prevented the low glucose-induced [Ca2+](i) oscillations in glucagon-releasing alpha-cells. The effects of 17beta-E2 on alpha cells were mimicked by the cGMP permeable analog 8bromo-cGMP and blocked by the cGMP-dependent protein kinase (PKG) inhibitor KT5823. Evidence indicated that these were membrane actions mediated by a nonclassical ER. Both effects were rapid in onset and were reproduced by 17beta-E2 linked to horseradish peroxidase, a cell-impermeable molecule. Furthermore, these actions were not blocked by the specific ER blocker ICI 182,780. Competition studies performed with 17beta-E2 linked to horseradish peroxidase binding in alpha-cells supported the idea that the membrane receptor involved is neither ERalpha nor ERbeta. Additionally, the binding site was shared by the neurotransmitters epinephrine, norepinephrine, and dopamine and had the same pharmacological profile as the receptor previously described for beta-cells. Therefore, rapid estrogen actions in islet cells are initiated by a nonclassical estrogen membrane receptor. PMID- 11875109 TI - Induction of human liver X receptor alpha gene expression via an autoregulatory loop mechanism. AB - The liver X receptors (LXRs), members of the nuclear receptor superfamily, play an important role in controlling lipid homeostasis by activating several genes involved in reverse cholesterol transport. These include members of the ATP binding cassette (ABC) superfamily of transporter proteins ABCA1 and ABCG1, surface constituents of plasma lipoproteins like apolipoprotein E, and cholesterol ester transport protein. They also play an important role in fatty acid metabolism by activating the sterol regulatory element-binding protein 1c gene. Here, we identify human LXRalpha (hLXRalpha) as an autoinducible gene. Induction in response to LXR ligands is observed in multiple human cell types including macrophages and occurs within 2--4 h. Analysis of the hLXRalpha promoter revealed three LXR response elements (LXREs); one exhibits strong affinity for both LXRalpha:RXR and LXRbeta:RXR (a type I LXRE), and deletion and mutational studies indicate it plays a critical role in LXR-mediated induction. The other two LXREs are identical to each other, exist within highly conserved Alu repeats, and exhibit selective binding to LXRalpha:RXR (type II LXREs). In transfections, the type I LXRE acts as a strong mediator of both LXRalpha and LXRbeta activity, whereas the type II LXRE acts as a weaker and selective mediator of LXRalpha activity. Our data suggest a model in which LXR ligands trigger an autoregulatory loop leading to selective induction of hLXRalpha gene expression. This would lead to increased hLXRalpha levels and transcription of its downstream target genes such as ABCA1, providing a simple yet exquisite mechanism for cells to respond to LXR ligands and cholesterol loading. PMID- 11875111 TI - Inhibition of androgen receptor (AR) function by the reproductive orphan nuclear receptor DAX-1. AB - DAX-1 (NROB1) is an atypical member of the nuclear receptor family that is predominantly expressed in mammalian reproductive tissues. While a receptor function of DAX-1 remains enigmatic, previous work has indicated that DAX-1 inhibits the activity of the orphan receptor steroidogenic factor 1 and the estrogen receptors (ERs), presumably via direct occupation of the coactivator binding surface and subsequent recruitment of additional corepressors. In vivo evidence points at a particular role of DAX-1 for the development and maintenance of male reproductive functions. In this study, we have identified the androgen receptor (AR) NR3C4 as a novel target for DAX-1. We show that DAX-1 potently inhibits ligand-dependent transcriptional activation as well as the interaction between the N- and C-terminal activation domains of AR. We provide evidence for direct interactions of the two receptors that involve the N-terminal repeat domain of DAX-1 and the C-terminal ligand-binding and activation domain of AR. Moreover, DAX-1, known to shuttle between the cytoplasm and the nucleus, is capable of relocalizing AR in both cellular compartments, suggesting that intracellular tethering is associated with DAX-1 inhibition. These results implicate novel inhibitory mechanisms of DAX-1 action with particular relevance for the modulation of androgen-dependent gene transcription in the male reproductive system. PMID- 11875113 TI - Regulation of the orphan nuclear receptor steroidogenic factor 1 by Sox proteins. AB - Steroidogenic factor 1 (SF-1) is an essential factor in endocrine proliferation and gene expression. Despite the fact that SF-1 expression is restricted to specialized cells within the endocrine system, the only identified regulatory factors of SF-1 are the ubiquitously expressed E-box proteins (upstream stimulatory factors 1 and 2). Sequence examination of the SF-1 proximal promoter revealed a conserved site of AACAAAG (Sox-BS1), which matches exactly the defined consensus Sox protein binding element. Among the approximately 20 known members of the Sox gene family, we focused on Sox3, Sox8, and Sox9, based on their coexpression with SF-1 in the embryonic testis. Indeed, all three of these Sox proteins were capable of binding the proximal Sox-BS1 within the SF-1 promoter ( 110 to -104), albeit with differing affinities. Of the three Sox proteins, Sox9 exhibited high-affinity binding to the Sox-BS1 element and consistently activated SF-1 promoter-reporter constructs. Mutating the Sox-BS1 attenuated SF-1 promoter activity in both embryonic and postnatal Sertoli cells, as well as in the adrenocortical cell line, Y1. Our findings, taken together with the overlapping expression profiles of Sox9 and SF-1, and the similar intersex phenotypes associated with both SOX9 and SF-1 human mutations, suggest that Sox9 up regulates SF-1 and accounts partially for the sexually dimorphic expression pattern of SF-1 observed during male gonadal differentiation. PMID- 11875114 TI - Neogenesis of beta-cells in adult BETA2/NeuroD-deficient mice. AB - BETA2/NeuroD, a basic helix-loop-helix transcription factor, is expressed in pancreatic endocrine cells during development and regulates insulin gene expression. We demonstrated previously that the endocrine pancreas of BETA2/NeuroD-deficient mice undergoes massive apoptosis and, consequently, animals die of diabetes shortly after birth. Here we show that a significant fraction of BETA2-deficient mice in a new genetic background can survive diabetes and live to adulthood through the process of beta-cell neogenesis. Morphometric examination indicates that pancreatic beta-, but not alpha-cell mass, was restored to a level comparable to that of wild-type animals. However, the newly formed islet cells cannot form mature islets of Langerhans, indicating an indispensable role of BETA2 in morphogenesis of normal islet structure. Furthermore, immunohistochemical examinations revealed that newly formed beta cells of BETA2/NeuroD-deficient mice come from two sources: either directly budding from the pancreatic ductal tree or from the preexisting beta-cells in the residual endocrine pancreas. Our results indicate that beta-cell neogenesis in our BETA2/NeuroD-deficient mice contributes to their survival, and these mice may provide a useful model for studying the mechanism of beta-cell regeneration. PMID- 11875115 TI - 14-3-3 facilitates insulin-stimulated intracellular trafficking of insulin receptor substrate 1. AB - The appearance of a complex between tyrosine-phosphorylated insulin receptor substrate 1 (IRS-1) and PI3K in a high-speed pellet fraction (HSP) is thought to be a key event in insulin action. Conversely, the disappearance of the IRS-1/PI3K complex from this fraction has been linked to insulin desensitization. The present study examines the role of 14-3-3, a specific phospho-serine binding protein, in mediating the disappearance of IRS-1 from the HSP after insulin treatment. An in vitro pull-down assay using recombinant 14-3-3 revealed that insulin enhances the association of 14-3-3 with IRS-1 in cultured adipocytes and that this is completely inhibited by wortmannin. An association of IRS-1 and 14-3 3 was also observed and was maximal after stimulation by insulin, when endogenous proteins were immunoprecipitated. Epidermal growth factor (EGF), 12-O tetradecanoylphorbol-13-acetate, and okadaic acid, other agents that cause serine/threonine phosphorylation of IRS-1, also stimulated IRS binding to 14-3-3. The enhancement of IRS-1 binding to 14-3-3 by insulin was accompanied by movement of IRS-1 and the p85 subunit of PI3K from the HSP to the cytosol. In keeping with a key role of 14-3-3 in mediating this redistribution of IRS-1, the complexes of IRS-1 and 14-3-3 were found in the cytosol but not in the HSP of insulin-treated cells. In addition, colocalization of IRS-1 and 14-3-3 was observed in the cytoplasm after insulin treatment by confocal microscopy. Finally, the addition of a phosphorylated 14-3-3 binding peptide to an adipocyte homogenate (to remove 14-3-3 from IRS-1) increased the abundance of IRS-1/PI3K complexes in the HSP and decreased their abundance in the cytosol. These findings strongly suggest that 14 3-3 participates in the intracellular trafficking of IRS-1 by promoting the displacement of serine-phosphorylated IRS-1 from particular structures. They also suggest that 14-3-3 proteins could play an integral role in the process of insulin desensitization. PMID- 11875116 TI - Jak2 is an essential tyrosine kinase involved in pregnancy-mediated development of mammary secretory epithelium. AB - The PRL receptor (PrlR) and the signal transducer and activator of transcription 5a (Stat5a) are essential for the proliferation and differentiation of mammary epithelium during pregnancy. Based on tissue culture cell experiments, Jak2 is the tyrosine kinase responsible for the phosphorylation of both the PrlR and Stat5. We have now used a genetic approach to test the role of Jak2 in the mammary gland, a PrlR-responsive tissue. Because Jak2-null embryos die at E12.5, we transplanted Jak2-null mammary anlagen into cleared fat pads of wild-type mice and investigated epithelial development during pregnancy. In the absence of Jak2, no secretory alveoli were present at parturition, and epithelial cell proliferation was reduced by 95% after an acute hormone treatment. Furthermore, the Na-K-Cl cotransporter, a ductal marker, was maintained in Jak2-null epithelium and the sodium-phosphate cotransporter type IIb, a secretory cell marker, was absent. Nuclear Stat5a was only observed in a few epithelial cells in Jak2-null glands at pregnancy and parturition compared with most epithelial cells in wild-type glands. Taken together, our results demonstrate that Jak2 is a critical tyrosine kinase that conveys intracellular signals necessary for proliferation and differentiation of mammary epithelium during pregnancy. PMID- 11875117 TI - Direct regulation of beta3-integrin subunit gene expression by HOXA10 in endometrial cells. AB - Estrogen and progesterone regulate HOXA10 expression in the endometrium, where HOXA10 is necessary for implantation. The integrins are also involved in early embryo-endometrial interactions. Here we show that HOXA10 directly regulates beta3-integrin subunit expression in the endometrium, likely mediating the effect of sex steroids on beta3-integrin expression. beta3-Integrin expression was decreased in endometrium shown to have low HOXA10 expression. beta3-Integrin mRNA levels were increased in endometrial adenocarcinoma cells (Ishikawa) transfected with pcDNA3.1/HOXA10, and decreased in cells treated with HOXA10 antisense. Seven consensus HOXA10 binding sites were identified 5' of the beta3-integrin gene. Direct binding of HOXA10 protein to four sites was demonstrated by EMSA. Reporter gene expression increased in BT-20 cells cotransfected with pcDNA3.1/ HOXA10 and pGL3-promoter vector containing region F (encompassing all seven HOXA10 consensus sites). A 41-bp segment (Region A) showed highest affinity binding to HOXA10 protein. Increased reporter expression, equal in magnitude to that obtained with Region F, was obtained with Region A. HOXA10 protein binding within Region A was localized by deoxyribonuclease I footprinting. beta3-Integrin expression was directly up-regulated by HOXA10 through a 41-bp 5'-regulatory element. Sex steroids regulate the expression of endometrial beta3-integrin through a pathway involving HOXA10. PMID- 11875118 TI - Expression of FKHR, FKHRL1, and AFX genes in the rodent ovary: evidence for regulation by IGF-I, estrogen, and the gonadotropins. AB - Follicular development is dependent on both intraovarian growth regulatory factors, such as IGF-I and estrogen, as well as the pituitary gonadotropins, FSH and LH. Recently, we have shown that FSH impacts the IGF-I pathway via stimulation of the PI3K cascade leading to phosphorylation of protein kinase B (PKB)/Akt and the PKB-related kinase, Sgk. This study was undertaken to determine if during ovarian follicular development FSH regulates putative targets of PKB and Sgk, namely specific Forkhead transcription factor family members. Using in vivo and in vitro mouse and rat models, we show 1) that FKHR [Forkhead homolog of rhabdomysarcoma = Forkhead box binding protein (Foxo1), FKHRL1 (Forkhead-like protein-1 = Foxo3), and AFX (a Forkhead transcription factor = Foxo4); all defined according to the Human and Mouse Gene Nomenclature Committee) are expressed in the rodent ovary and 2) that FSH regulates transcription of the FKHR gene as well as phosphorylation of FKHR protein. Specifically, FSH/PMSG (primarily via E2) enhance expression of the FKHR gene in granulosa cells of developing follicles. Furthermore, E2 enhances expression of other IGF-I pathway components (IGF-1Rbeta and Glut-1), and IGF-I enhances expression of ERbeta, indicating that these two hormones comprise an autocrine regulatory network within growing follicles. In contrast, FSH and LH/human CG (via cAMP, PKA, and PI3K pathways) terminate FKHR expression as granulosa cells differentiate to luteal cells. In naive granulosa cells, both FSH and IGF-I stimulate rapid phosphorylation of FKHR at multiple sites causing its redistribution from the nucleus to the cytoplasm in a PI3K-dependent manner. However, the effects of FSH and IGF-I differ markedly in differentiated granulosa cells in which FSH (but not IGF-I) induces Sgk and enhances phosphorylation of FKHR, PKB, and Sgk. The elevated expression of FKHR in granulosa cells of growing follicles indicates that FKHR may be linked to the proliferation of granulosa cells and that its phosphorylation by FSH, IGF-I, and other factors may impact its functional activity in this process. Thus, as a target of FSH (cAMP), E2 and IGF-I signaling in granulosa cells, FKHR likely coordinates numerous cell survival mechanisms. PMID- 11875119 TI - A single residue (arg46) located within the N-terminus of the V1a vasopressin receptor is critical for binding vasopressin but not peptide or nonpeptide antagonists. AB - A fundamental issue in molecular endocrinology is to define how agonist:receptor interaction differs from antagonist:receptor interaction. The vasopressin V1a receptor (V1aR) is a member of a subfamily of related G protein-coupled receptors that are activated by the hormone AVP or related peptides. The N-terminus of the V1aR has recently been shown to be critical for binding agonists but not antagonists. Using a combination of N-terminally truncated constructs and alanine scanning mutagenesis, individual residues that provide these agonist-specific binding epitopes have now been identified in this study. Our data establish that a single residue, Arg46, is critical for AVP binding to the V1aR. Systematic substitution revealed that Arg was required at this locus and could not be substituted by Lys, Glu, Leu, or Ala. In contrast, antagonist binding (cyclic or linear, peptide or nonpeptide) was unaffected. Disruption of Arg46 also resulted in defective intracellular signaling. Arginine is conserved at this locus in all members of the neurohypophysial peptide hormone receptor family cloned to date, indicative of a fundamental role in receptor function. In addition to Arg46, the residues Leu42, Gly43, Asp45 form a patch contributing to AVP binding. This study provides molecular insight into the role of the V1aR N-terminus and key differences between agonist and antagonist binding requirements. PMID- 11875120 TI - Independence of angiotensin II-induced MAP kinase activation from angiotensin type 1 receptor internalization in clone 9 hepatocytes. AB - The agonist-induced internalization of several G protein-coupled receptors is an obligatory requirement for their activation of MAPKs. Studies on the relationship between endocytosis of the angiotensin II (Ang II) type 1 receptor (AT1-R) and Ang II-induced ERK1/2 activation were performed in clone 9 (C9) rat hepatic cells treated with inhibitors of endocytosis [sucrose, phenylarsine oxide (PAO), and concanavalin A]. Although Ang II-induced endocytosis of the AT1-R was prevented by sucrose and PAO, and was partially inhibited by concanavalin A, there was no impairment of Ang II-induced ERK activation. However, the specific epidermal growth factor receptor (EGF-R) kinase inhibitor, AG1478, abolished Ang II-induced activation of ERK1/2. Sucrose and PAO also inhibited EGFinduced internalization of the EGF-R in C9 cells, and the inability of these agents to impair EGF-induced ERK activation suggested that the latter is also independent of receptor endocytosis. In COS-7 cells transiently expressing the rat AT1A-R, Ang II also caused ERK activation through EGF-R transactivation. Furthermore, a mutant AT1A-R with truncated carboxyl terminus and impaired internalization retained full ability to activate ERK1/2 in response to Ang II stimulation. These findings demonstrate that Ang II-induced ERK1/2 activation in C9 hepatocytes is independent of both AT1-R and EGF-R endocytosis and is mediated by transactivation of the EGF-R. PMID- 11875121 TI - TASK-3 dominates the background potassium conductance in rat adrenal glomerulosa cells. AB - In a preceding study we showed that the highly negative resting membrane potential of rat adrenal glomerulosa cells is related to background potassium channel(s), which belong to the two-pore domain channel family. TWIK-related acid sensitive K+ channel (TASK-1) expression was found in glomerulosa tissue, and the currents elicited by injection of glomerulosa mRNA (I(glom)) or TASK-1 cRNA (I(TASK-1)) showed remarkable similarity in Xenopus laevis oocytes. However, based on the different sensitivity of these currents to acidification, we concluded that TASK-1 may be responsible for a maximum of 25% of the weakly pH dependent glomerulosa background K+ current. Here we demonstrate that TASK-3, a close relative of TASK-1, is expressed abundantly in glomerulosa cells. Northern blot detected TASK-3 message in adrenal glomerulosa, but not in other tissues. Quantitative RT-PCR experiments indicated even higher mRNA expression of TASK-3 than TASK-1 in glomerulosa tissue. Similarly to the glomerulosa background current, the current expressed by injection of TASK-3 cRNA (I(TASK-3)) was less acid-sensitive than I(TASK-1). Ruthenium red in the micromolar range inhibited I(glom) and I(TASK-3), but not I(TASK-1). Like I(TASK-1), I(TASK-3) was inhibited by stimulation of AT1a angiotensin II receptor coexpressed with the potassium channel. The high level of expression and its pharmacological properties suggest that TASK-3 dominates the resting potassium conductance of glomerulosa cells. PMID- 11875122 TI - Mutation of the Calpha subunit of PKA leads to growth retardation and sperm dysfunction. AB - The intracellular second messenger cAMP affects cell physiology by directly interacting with effector molecules that include cyclic nucleotide-gated ion channels, cAMP-regulated G protein exchange factors, and cAMP-dependent protein kinases (PKA). Two catalytic subunits, Calpha and Cbeta, are expressed in the mouse and mediate the effects of PKA. We generated a null mutation in the major catalytic subunit of PKA, Calpha, and observed early postnatal lethality in the majority of Calpha knockout mice. Surprisingly, a small percentage of Calpha knockout mice, although runted, survived to adulthood. This growth retardation was not due to decreased GH production but did correlate with a reduction in IGF I mRNA in the liver and diminished production of the major urinary proteins in kidney. The survival of Calpha knockout mice after birth is dependent on the genetic background as well as environmental factors, but sufficient adult animals were obtained to characterize the mutants. In these animals, compensatory increases in Cbeta levels occurred in brain whereas many tissues, including skeletal muscle, heart, and sperm, contained less than 10% of the normal PKA activity. Analysis of sperm in Calpha knockout males revealed that spermatogenesis progressed normally but that mature sperm had defective forward motility. PMID- 11875124 TI - Endocrine-related resources from the National Institutes of Health. PMID- 11875125 TI - Hydrogenases and hydrogen metabolism of cyanobacteria. AB - Cyanobacteria may possess several enzymes that are directly involved in dihydrogen metabolism: nitrogenase(s) catalyzing the production of hydrogen concomitantly with the reduction of dinitrogen to ammonia, an uptake hydrogenase (encoded by hupSL) catalyzing the consumption of hydrogen produced by the nitrogenase, and a bidirectional hydrogenase (encoded by hoxFUYH) which has the capacity to both take up and produce hydrogen. This review summarizes our knowledge about cyanobacterial hydrogenases, focusing on recent progress since the first molecular information was published in 1995. It presents the molecular knowledge about cyanobacterial hupSL and hoxFUYH, their corresponding gene products, and their accessory genes before finishing with an applied aspect--the use of cyanobacteria in a biological, renewable production of the future energy carrier molecular hydrogen. In addition to scientific publications, information from three cyanobacterial genomes, the unicellular Synechocystis strain PCC 6803 and the filamentous heterocystous Anabaena strain PCC 7120 and Nostoc punctiforme (PCC 73102/ATCC 29133) is included. PMID- 11875126 TI - Cytoskeleton of apicomplexan parasites. AB - The Apicomplexa are a phylum of diverse obligate intracellular parasites including Plasmodium spp., the cause of malaria; Toxoplasma gondii and Cryptosporidium parvum, opportunistic pathogens of immunocompromised individuals; and Eimeria spp. and Theileria spp., parasites of considerable agricultural importance. These protozoan parasites share distinctive morphological features, cytoskeletal organization, and modes of replication, motility, and invasion. This review summarizes our current understanding of the cytoskeletal elements, the properties of cytoskeletal proteins, and the role of the cytoskeleton in polarity, motility, invasion, and replication. We discuss the unusual properties of actin and myosin in the Apicomplexa, the highly stereotyped microtubule populations in apicomplexans, and a network of recently discovered novel intermediate filament-like elements in these parasites. PMID- 11875127 TI - Molecular biologist's guide to proteomics. AB - The emergence of proteomics, the large-scale analysis of proteins, has been inspired by the realization that the final product of a gene is inherently more complex and closer to function than the gene itself. Shortfalls in the ability of bioinformatics to predict both the existence and function of genes have also illustrated the need for protein analysis. Moreover, only through the study of proteins can posttranslational modifications be determined, which can profoundly affect protein function. Proteomics has been enabled by the accumulation of both DNA and protein sequence databases, improvements in mass spectrometry, and the development of computer algorithms for database searching. In this review, we describe why proteomics is important, how it is conducted, and how it can be applied to complement other existing technologies. We conclude that currently, the most practical application of proteomics is the analysis of target proteins as opposed to entire proteomes. This type of proteomics, referred to as functional proteomics, is always driven by a specific biological question. In this way, protein identification and characterization has a meaningful outcome. We discuss some of the advantages of a functional proteomics approach and provide examples of how different methodologies can be utilized to address a wide variety of biological problems. PMID- 11875129 TI - Regulation of cellular differentiation in filamentous cyanobacteria in free living and plant-associated symbiotic growth states. AB - Certain filamentous nitrogen-fixing cyanobacteria generate signals that direct their own multicellular development. They also respond to signals from plants that initiate or modulate differentiation, leading to the establishment of a symbiotic association. An objective of this review is to describe the mechanisms by which free-living cyanobacteria regulate their development and then to consider how plants may exploit cyanobacterial physiology to achieve stable symbioses. Cyanobacteria that are capable of forming plant symbioses can differentiate into motile filaments called hormogonia and into specialized nitrogen-fixing cells called heterocysts. Plant signals exert both positive and negative regulatory control on hormogonium differentiation. Heterocyst differentiation is a highly regulated process, resulting in a regularly spaced pattern of heterocysts in the filament. The evidence is most consistent with the pattern arising in two stages. First, nitrogen limitation triggers a nonrandomly spaced cluster of cells (perhaps at a critical stage of their cell cycle) to initiate differentiation. Interactions between an inhibitory peptide exported by the differentiating cells and an activator protein within them causes one cell within each cluster to fully differentiate, yielding a single mature heterocyst. In symbiosis with plants, heterocyst frequencies are increased 3- to 10-fold because, we propose, either differentiation is initiated at an increased number of sites or resolution of differentiating clusters is incomplete. The physiology of symbiotically associated cyanobacteria raises the prospect that heterocyst differentiation proceeds independently of the nitrogen status of a cell and depends instead on signals produced by the plant partner. PMID- 11875131 TI - Delayed childbearing and its impact on population rate changes in lower birth weight, multiple birth, and preterm delivery. AB - OBJECTIVE: This study quantified the impact of delayed childbearing (maternal age greater-than-or-equal to 35 years) on population rate changes in low birth weight (LBW; < 2500 g), preterm delivery (< 37 weeks), multiple births, and small for gestational age (SGA; < 10th percentile) in Alberta, Canada, between 1990 (N = 42 930) and 1996 (N = 37 710). METHODS: Data were obtained from the provincial notification of a live or stillbirth. Analyses included relative risk estimates and chi(2) tests for trend. Potential confounding attributable to in vitro fertilization was investigated. RESULTS: The proportion of births to women greater-than-or-equal to 35 years of age was 8.4% in 1990 and 12.6% in 1996, a 51.2% increase. Among these women, LBW delivery increased 11%, and preterm delivery increased 14%. Delayed childbearing accounted for 78% of the change in LBW rate in the population and 36% of the change in preterm delivery rate in the population. Provincial multiple birth rates increased by 15% for twins and 14% for triplets. Delayed childbearing accounted for 15% of the twin increase and 69% of the triplet increase. When in vitro fertilization pregnancies were excluded, the change was 43% for preterm rates, 100% for LBW, 14% for twins, and 9% for triplets. Delayed childbearing did not contribute to changes in singleton SGA deliveries. CONCLUSIONS: The findings suggest that the recent increase in LBW and preterm delivery is partly related to the population phenomenon of delayed childbearing. Maternal age was not related to changes in SGA, suggesting that the age effect is through pregnancy complications that lead to preterm delivery and LBW. Prospective parents should be informed about the higher risk for neonatal morbidity associated with delayed childbearing. Health care providers should be aware of the impact of delayed childbearing on health care resources. PMID- 11875132 TI - Pediatric resident training in the diagnosis and treatment of acute otitis media. AB - OBJECTIVE: To investigate the presence and characteristics of formalized curricula for pediatric resident education for the diagnosis and treatment of acute otitis media. DESIGN: A survey was mailed to the program directors of all 224 accredited US and Canadian pediatric residency programs. Questions focused on the development, components, and teaching of any otitis media curriculum. Program directors were also questioned about their approach towards evaluating resident competency of otitis media and their future plans for otitis media-related training. RESULTS: A total of 144 program directors (64%) responded to the survey. Among respondents, 59% had some form of formalized education related to the diagnosis or treatment of otitis media. These curricula primarily consisted of lectures by general pediatricians <3 times per year. CONCLUSIONS: Although otitis media is the most common disease seen by practicing general pediatricians, the Residency Review Committee in Pediatrics does not mandate formal otitis media related training. Aside from informal case-by-case education, this survey demonstrated that only slightly more than half of all pediatric residency programs have some formalized resident education of this common pediatric problem, and most of those curricula are infrequent lectures. PMID- 11875133 TI - A new look at myelomeningoceles: functional level, vertebral level, shunting, and the implications for fetal intervention. AB - OBJECTIVE: Previous reports have suggested that 80% to 85% of patients who have a myelomeningocele (MMC) and undergo surgical repair after birth develop hydrocephalus and require the placement of a ventricular shunt. However, the rate of shunting as a function of spinal level is not well established. We sought to determine the distribution of postnatally repaired MMC lesions as characterized by both functional and radiologic assessment, as well as the incidence of shunting when patients were categorized according to these 2 methods. METHODS: A retrospective chart review of 297 patients who were born with open MMCs and followed in the spina bifida clinic at the Children's Hospital of Philadelphia was performed. The presence or absence of a shunt was determined for each patient. Functional spinal level was determined by the best-recorded neurologic examination and vertebral level by spine radiographs. RESULTS: The overall rate of ventricular shunting was 81%. The level of the lesion significantly affected the incidence of shunting, with more cephalad lesions correlating with higher rates. This was true both for functional and radiologic categorizations. A significantly higher shunt rate was found among patients with sacral lesions when categorized by radiologic rather than functional criteria. In 86% of patients, the functional level was found to be equal to or higher (worse) than the radiologic level. CONCLUSIONS: This study describes the natural history of ventricular shunting in MMC patients with relation to both radiologic and functional criteria. Fetal MMC closure is being performed in some centers in an attempt to decrease the incidence of shunting and to improve leg function in selected patients. The present data may serve as a comparison group and aid in the design and analysis of a prospective trial to assess the efficacy of this new procedure. PMID- 11875128 TI - Alpha-crystallin-type heat shock proteins: socializing minichaperones in the context of a multichaperone network. AB - Alpha-crystallins were originally recognized as proteins contributing to the transparency of the mammalian eye lens. Subsequently, they have been found in many, but not all, members of the Archaea, Bacteria, and Eucarya. Most members of the diverse alpha-crystallin family have four common structural and functional features: (i) a small monomeric molecular mass between 12 and 43 kDa; (ii) the formation of large oligomeric complexes; (iii) the presence of a moderately conserved central region, the so-called alpha-crystallin domain; and (iv) molecular chaperone activity. Since alpha-crystallins are induced by a temperature upshift in many organisms, they are often referred to as small heat shock proteins (sHsps) or, more accurately, alpha-Hsps. Alpha-crystallins are integrated into a highly flexible and synergistic multichaperone network evolved to secure protein quality control in the cell. Their chaperone activity is limited to the binding of unfolding intermediates in order to protect them from irreversible aggregation. Productive release and refolding of captured proteins into the native state requires close cooperation with other cellular chaperones. In addition, alpha-Hsps seem to play an important role in membrane stabilization. The review compiles information on the abundance, sequence conservation, regulation, structure, and function of alpha-Hsps with an emphasis on the microbial members of this chaperone family. PMID- 11875134 TI - Use of an Internet-based community surveillance network to predict seasonal communicable disease morbidity. AB - OBJECTIVES: We designed an Internet-based surveillance network that linked community clinic diagnoses with viral isolation rates and admission patterns at a related children's hospital. We hypothesized that community surveillance would successfully predict subsequent hospital admissions and laboratory viral isolations. Secondarily, we expected the network to monitor trends in disease and that posting this information on a Web site would be useful to physicians in daily practice. STUDY DESIGN: Data were collected from December 1999 through August 2000. Information was summarized and posted weekly on a Web site. Active public piloting of the site took place during August 2000, after which the project was evaluated through an electronic mail survey. The predictive ability of the community surveillance data was evaluated by multivariate linear regression. RESULTS: Increases in the community diagnosis of most syndromes under surveillance, including lower respiratory infections (adjusted R(2) = 0.7086) and gastroenteritis (adjusted R(2) = 0.6532) successfully predicted an increase in subsequent hospital admissions. Community surveillance also successfully predicted laboratory isolation of associated viral organisms. Physicians completing the evaluation (N = 11) indicated that the site provided information useful in daily practice for both physician and parent education. CONCLUSIONS: An Internet-based surveillance network linking a hospital with community physicians is beneficial to the hospital in predicting waves of severe cases requiring admission and reciprocally provides useful information to physicians in daily practice regarding the incidence and cause of seasonal disease in the community. PMID- 11875135 TI - Internet use in families with children requiring cardiac surgery for congenital heart disease. AB - OBJECTIVE: We sought to document the prevalence of Internet access and usage patterns among families who have children with congenital heart disease presenting for cardiac surgery. DESIGN: A cross-sectional study was performed utilizing a questionnaire (10 questions) as the study tool. Statistical analysis was performed on all completed questionnaires. The chi(2) test was used to evaluate categorical variables and the t test to evaluate continuous variables. RESULTS: Two hundred seventy-five questionnaires were completed and analyzed. Thirty-seven percent (102/275) of the children had a cyanotic congenital heart defect. There were 21 children with Trisomy 21. Fifty-eight percent (160/275) of families had access to the Internet. The most common locations for accessing the Internet were home (80%; 129/160) and work (51%; 82/160). There were no significant differences in Internet access with regards to underlying individual congenital heart defect, cyanotic versus acyanotic heart defects, or congenital heart defects with functional univentricular hearts versus biventricular hearts. Families with older children (12--24 years) were more likely to have Internet access. Families of children undergoing placement of a right ventricle to pulmonary artery conduit were more likely to have Internet access. Of the 160 families with Internet access, 58% (93/160) used the Internet to obtain information related to their child's cardiac diagnosis. Eighty-two percent (76/93) characterized locating cardiology-related information as easy. Six parents created interactive personal Web sites specifically related to their child's congenital heart defect. Although families with older patients (12--24 years) were more likely to have access to the Internet, this did not translate into greater use of the Internet to obtain cardiology-related information. Among families who accessed the Internet for cardiology-related information, 95% (88/93) of families characterized the information as helpful or very helpful in furthering the understanding of their child's heart defect. CONCLUSION: Families are utilizing the Internet to educate themselves about congenital heart disease. Most parents consider the process easy and the information obtained helpful to the understanding of their child's congenital heart defect and surgery. Internet use in this patient population is expected to increase. Our vigilance in providing accurate Internet references, as well as in identifying inaccurate Internet information available to our patients and their parents, is of paramount importance. PMID- 11875136 TI - Impact of necrotizing enterocolitis on length of stay and hospital charges in very low birth weight infants. AB - OBJECTIVE: To determine the impact of necrotizing enterocolitis (NEC) on length of stay and hospital charges. DESIGN: Case-control study. SETTING: Two neonatal intensive care units in an academic medical center. PATIENTS: Infants born in 1992--1994 with birth weight <1500 g, matched by gestational age, hospital, and month of birth. MEASUREMENTS AND MAIN RESULTS. We performed odds ratio and t testing. As with previous studies, there was no single factor that increased the risk for developing NEC. However, the diagnosis of NEC increased the risk for death, infection, and the need for central line placement. Infants with surgical NEC had lengths of stay that exceeded those of controls by 60 days, whereas lengths of stay among infants with medical NEC exceeded those of controls by 22 days. Based on length of stay, the estimated total hospital charges for infants with surgical NEC averaged $186 200 in excess of those for controls and $73 700 more for infants with medical NEC. The yearly additional hospital charges for NEC were $6.5 million or $216 666 per survivor. CONCLUSIONS: A diagnosis of NEC in the very low birth weight infant imposes a significant additional financial burden to the individual patient as well as the neonatal community as a whole. This expense justifies additional research into preventive measures and potentially costly therapies aimed at reducing the incidence of NEC. These data also provide an estimated cost to compare the cost effectiveness of new preventive measures for NEC. PMID- 11875137 TI - Self-esteem of adolescents who were born prematurely. AB - OBJECTIVE: To determine whether there are any differences between the self-esteem of a cohort of adolescents who were extremely low birth weight (ELBW) in comparison with term controls (Cs); and to test the associations between self esteem and several predefined predictor variables. BACKGROUND: Self-esteem is considered to play a significant role in psychological adjustment and scholastic success. Little information exists on how adolescents who were ELBW regard themselves. DESIGN/METHODS: Longitudinal follow-up of a regional cohort of 132/169 (78%) ELBW survivors and 127/145 (88%) sociodemographically matched Cs, born between 1977 and 1982. MEASURES: Harter Adolescent Self-Perception Profile (1988) with 9 dimensions, including Global Self-Worth, socioeconomic status (Hollingshead), height-for-age and weight-for-age z scores, and Wide Range Achievement Test---Revised (WRAT-R; Reading, Spelling, and Arithmetic). DATA ANALYSIS: General linear model multiple analyses of covariance were performed to determine whether significant relationships existed between the 9 self-esteem dimensions and the independent variables of birth weight status and gender, and the covariates of age, socioeconomic status, physical development, and academic achievement. RESULTS: Global Self Worth was similar for ELBW and Cs (means: 3.1 and 3.2). Multivariate effects revealed no interactions, but significant main effects emerged for birth weight status, gender, weight-for-age z scores, age in months, and for all 3 WRAT-R subtests, all effect sizes medium to large. Follow up analysis of covariance revealed medium-size gender effects for athletic competence (means: 3.1 and 2.6), and physical appearance (means: 2.9 and 2.5), where boys rated themselves significantly higher on both domains; and age effects, where older teens rated themselves better for job competence. Significant but small effect sizes emerged for the following: 1) weight-for-age z scores, where heavier youth rated themselves higher on close friendships, 2) gender, where girls had higher ratings for close friendships, 3) birth weight, where Cs rated themselves higher on athletic competence, and 4) WRAT-R math effect, where children with higher math scores rated themselves better on scholastic competence. CONCLUSIONS: Overall, ELBW teenagers do not differ significantly from C teenagers on most dimensions of self-esteem. Gender effects emerged on some Harter domains. PMID- 11875138 TI - Noninvasive continuous monitoring of the effects of head position on brain hemodynamics in ventilated infants. AB - HYPOTHESIS: Laying supine with the head in midline position improves cerebral venous return by preventing functional occlusion of the vessels of the neck. OBJECTIVES: To assess changes in cerebral blood volume (DeltaCBV) and cerebral blood flow (CBF) with the position of the head in ventilated patients using a noninvasive method. The influence of the type of ventilation and birth weight was evaluated. METHODS: Thirteen conventionally ventilated and 8 high-frequency oscillatory ventilated infants, with mean gestational ages and birth weights of 31 +/- 5 weeks (24--38) and 1575 +/- 803 g (560--3000), respectively, were studied 5.8 +/- 7.8 days (1--33) after birth. DeltaCBV (mL/100 g) and CBF (mL/100 g/min) were measured by near-infrared spectroscopy with the head in supine midline position (DeltaCBVs, CBFs) and rotated 90 to one side (DeltaCBVlat, CBFlat). Heart rate, peripheral saturation, transcutaneous PCO(2), and blood pressure were monitored continuously. Ventilatory settings remained constant throughout the study period. RESULTS: Mean DeltaCBVs was lower than mean DeltaCBVlat, although no changes in blood pressure, transcutaneous PCO(2), oxygenation, or heart rate occurred. This change in DeltaCBV was not associated with the type of ventilation or birth weight, but the differences tended to be greater (dDeltaCBV = DeltaCBVlat-DeltaCBVs) in the smallest infants (<1200 g). In contrast, CBF did not vary. CONCLUSION: The supine midline position of the head favors cerebral venous drainage and helps to prevent elevation of CBV. SPECULATION: This finding may be important in the first days of life, particularly in tiny preterm infants recovering from lung disease with improving lung compliance, in which functional obstruction of cerebral venous drainage should be avoided. PMID- 11875130 TI - Secretory pathway of trypanosomatid parasites. AB - The Trypanosomatidae comprise a large group of parasitic protozoa, some of which cause important diseases in humans. These include Trypanosoma brucei (the causative agent of African sleeping sickness and nagana in cattle), Trypanosoma cruzi (the causative agent of Chagas' disease in Central and South America), and Leishmania spp. (the causative agent of visceral and [muco]cutaneous leishmaniasis throughout the tropics and subtropics). The cell surfaces of these parasites are covered in complex protein- or carbohydrate-rich coats that are required for parasite survival and infectivity in their respective insect vectors and mammalian hosts. These molecules are assembled in the secretory pathway. Recent advances in the genetic manipulation of these parasites as well as progress with the parasite genome projects has greatly advanced our understanding of processes that underlie secretory transport in trypanosomatids. This article provides an overview of the organization of the trypanosomatid secretory pathway and connections that exist with endocytic organelles and multiple lytic and storage vacuoles. A number of the molecular components that are required for vesicular transport have been identified, as have some of the sorting signals that direct proteins to the cell surface or organelles in the endosome-vacuole system. Finally, the subcellular organization of the major glycosylation pathways in these parasites is reviewed. Studies on these highly divergent eukaryotes provide important insights into the molecular processes underlying secretory transport that arose very early in eukaryotic evolution. They also reveal unusual or novel aspects of secretory transport and protein glycosylation that may be exploited in developing new antiparasite drugs. PMID- 11875139 TI - Genetic screening for maternal uniparental disomy of chromosome 7 in prenatal and postnatal growth retardation of unknown cause. AB - OBJECTIVE: Many short-statured children lack an etiologic explanation for their retarded growth. Recently, uniparental disomy (UPD), the inheritance of both chromosomes of a chromosome pair from only 1 parent, has been associated with short stature for many chromosomes. Silver-Russell syndrome (SRS) represents an extreme syndrome of intrauterine growth retardation (IUGR) and slight dysmorphic signs, and maternal UPD of human chromosome 7 (matUPD7) has been observed in approximately 10% of SRS cases. In addition, matUPD7 has been reported in patients with only slight dysmorphic features and prenatal or postnatal growth retardation. The objectives of this study were to study the role of matUPD7 in growth failure of unknown cause and in cases of SRS, and to evaluate the efficiency of genetic testing for matUPD7 as a diagnostic tool. METHODS: DNA samples were studied from 205 children, 92 girls and 113 boys, with short stature of unknown cause and their parents. The patient cohort included 39 cases of SRS, 91 patients with IUGR and subsequent postnatal short stature, and 75 patients with postnatal growth retardation only. MatUPD7 was screened for by genotyping DNA samples from the patient, mother, and father with 13 chromosome-7-specific polymorphic microsatellite markers. RESULTS: Six (3%) of 205 matUPD7 cases were observed exclusively among 39 (15%) SRS patients studied. Patients with IUGR and/or postnatal growth retardation and with dysmorphic features did not reveal cases of matUPD7. CONCLUSIONS: Our results indicate that matUPD7 cases are predominantly observed among patients meeting the criteria of SRS, and matUPD7 is not a common cause for growth retardation. Genetic screening for cases of matUPD7 among growth-retarded patients should be focused on patients with severe IUGR and features of SRS. In addition, matUPD7 screening is advisable in individuals with cystic fibrosis and other recessive disorders mapped to chromosome 7 who have unusually short stature. PMID- 11875140 TI - Inattention, hyperactivity, and symptoms of sleep-disordered breathing. AB - OBJECTIVE: Inattention and hyperactivity are frequent among children with sleep disordered breathing (SDB) and often improve when SDB is treated. However, the frequency of SDB symptoms among inattentive and hyperactive children has received little study. DESIGN: Cross-sectional survey. SETTING: Two university-affiliated but community-based general pediatrics clinics. PATIENTS: Patients consisted of N = 866 children (469 boys), aged 2.0 to 13.9 years (mean: 6.8 plus minus 3.2 years), with clinic appointments. MEASURES: A validated Pediatric Sleep Questionnaire assessed for habitual snoring (1 item), snoring severity (a 4-item subscale), sleepiness (4 items), and overall risk of SDB (16 items). Parents also completed 2 common behavioral measures, an inattention/hyperactivity scale (IHS) derived from the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition, and the hyperactivity index (HI, expressed as a t score) of the Conners' Parent Rating Scale. RESULTS: Habitual snoring was reported in 16% (95% confidence interval [CI]: 13, 19) of the participants. High HI scores (>60) were found in 13% (95% CI: 11, 16) of all participants, 22% (95% CI: 15, 29) of habitual snorers, and 12% (95% CI: 9, 14) of nonsnorers. Odds ratios between HI >60 and each of the following were: habitual snoring, 2.2 (95% CI: 1.4, 3.6); 1 additional positive symptom-item on the snoring scale, 1.3 (95% CI: 1.1, 1.5); 1 additional positive item on the sleepiness scale, 1.6 (95% CI: 1.4, 2.0); and a 1 standard deviation increase in the overall SDB score, 1.7 (95% CI: 1.4, 2.0; all odds ratios age- and sex-adjusted). Results were similar for high IHS scores (>1.25). Stratification by age and sex showed that most of the association with snoring (but not sleepiness) derived from boys <8 years old. CONCLUSIONS: Inattention and hyperactivity among general pediatric patients are associated with increased daytime sleepiness and---especially in young boys---snoring and other symptoms of SDB. If sleepiness and SDB do influence daytime behavior, the current results suggest a major public health impact. PMID- 11875141 TI - The significance of gastric residuals in the early enteral feeding advancement of extremely low birth weight infants. AB - OBJECTIVE: To evaluate whether the mean gastric residual volume (GRV) and green gastric residuals (GR) themselves are significant predictors of feeding intolerance in the early enteral feeding advancement in extremely low birth weight (ELBW; <1000 g) infants. DESIGN: Ninety-nine ELBW infants were fed following a standardized protocol (day 3--14). At 48 hours of age, milk feeding was started (12 mL/kg/d increments, 12 meals per day). GR were checked before each feeding, and a GRV up to 2 mL/3 mL in infants less-than-or-equal750 g/>750 g was tolerated. In cases of increased GRV, feedings were reduced or withheld. The color of GR was assessed as clear, milky, green-clear, green-cloudy, blood stained, or hemorrhagic. Multiple regression analysis was used to study the effect of the mean GRV and the color of GR on the feeding volume on day 14 (V14). RESULTS: The median V14 was 103 mL/kg/d (0--166). V14 increased with an increasing percentage of milky GR, whereas the mean GRV and the color green did not have a significant effect. CONCLUSIONS: 1) Early enteral feeding could be established in ELBW infants. The critical GRV seems to be above 2 mL/3 mL because there was no significant negative correlation between the mean GRV and V14. 2) Green GR were not negatively correlated with V14 and should not slow down the advancement of feeding volumes in absence of other clinical signs and symptoms. PMID- 11875142 TI - The usefulness of children's drawings in the diagnosis of headache. AB - OBJECTIVE: To determine whether drawings can aid in the differential diagnosis of headaches in children. METHODS: Before taking any history, 226 children who were seen consecutively for the evaluation of headache were asked to draw a picture to show how their headache felt. The pictures were then scored as migraine or nonmigraine by pediatric neurologists who were blinded to the clinical history. A clinical diagnosis of headache type was determined independently by another pediatric neurologist using the usual history and examination. The diagnoses of headache type based on the pictures drawn and the clinical findings obtained were then compared to calculate the sensitivity, specificity, and predictive values of the drawings for the diagnosis of migraine. RESULTS: Children produced dramatic and insightful headache drawings. Compared with the clinical diagnosis (gold standard), headache drawings had a sensitivity of 93.1%, a specificity of 82.7%, and a positive predictive value (PPV) of 87.1% for migraine. That is, drawings that contained an artistic feature consistent with migraine (eg, pounding pain, nausea/vomiting, desire to lie down, periorbital pain, photophobia, visual scotoma) predicted the clinical diagnosis of migraine in 87.1% of cases. Predictive values were also calculated for specific migraine-associated features on drawings: artistic depiction of focal neurologic signs, periorbital pain, recumbency, visual symptoms (photophobia, scotomata), or nausea/vomiting had a PPV of >90% for migraine; severe or pounding pain had a PPV of >80% for migraine. Band-like pain was not predictive of migraine (PPV of 11.1%). Features on drawings such as sadness or crying did not differentiate migraine from nonmigraine headaches. CONCLUSIONS: Children's headache drawings are a simple, inexpensive aid in the diagnosis of headache type, with a very high sensitivity, specificity, and predictive value for migraine versus nonmigraine headaches. We encourage the use of drawings in the evaluation of any child with a headache, as an adjunct to the clinical history and physical examination. PMID- 11875143 TI - Hypospadias and early gestation growth restriction in infants. AB - OBJECTIVE: There has been a major increase in the incidence of hypospadias in infants in the 1990s, but the risk factors are not known. Although there are scattered reports in the literature regarding the association of low birth weight and hypospadias, this has not been systematically studied. The objective of this study was to determine the association between early gestation intrauterine growth and hypospadias. METHODS: A retrospective review of 13 years of admissions to 2 tertiary care neonatal intensive care units (NICUs) in Connecticut (1987- 2000) showed that 112 (1.66%) of 6746 male infants had any degree of hypospadias. Of these, 8 were part of a genetic syndrome and were excluded. A retrospective cohort analysis of these 6738 infants was performed. Infant growth parameters at birth (weight, head circumference, and length) were analyzed along with maternal risk factors known to be associated with changes in fetal growth, including maternal age, race, diagnosis of preeclampsia, gestational diabetes, and maternal use of alcohol or tobacco or substance abuse during pregnancy. RESULTS: The incidence of hypospadias in the NICU population increased 10-fold from 0.4% in 1987 to 4% in the first quarter of 2000. Hypospadias was significantly more common in infants who had uniformly poor intrauterine growth (<10th percentiles) in the various parameters measured: birth weight, length, or head circumference. There were no significant differences in maternal age or race, nor were there differences in the use of alcohol, tobacco, or street drugs by the mother. There were no differences between singletons and multiple-gestation births. However, the frequency of occurrence was significantly higher among first-born infants (1.9%) compared with all other infants (0.9%). CONCLUSIONS: The incidence of hypospadias in our NICU population has increased 10-fold during the 13-year period of study. There was a significant association of hypospadias with poor intrauterine growth. The growth restriction was probably of early gestational cause as there was proportionate involvement of somatic (weight and length) and brain growth (head circumference). The increasing frequency of hypospadias and its association with poor intrauterine growth originating in early gestation suggests that common environmental factor(s) that have an impact on both conditions may be involved. PMID- 11875144 TI - Stature at time of diagnosis of type 1 diabetes mellitus. AB - OBJECTIVE: To assess the stature of children with type 1 diabetes mellitus at diagnosis. METHODS: We collected data from 451 records of children who were examined in a pediatric diabetes clinic and used data from the Third National Health and Nutrition Examination Survey for 10 522 children as control group. Analytical techniques included linear and logistic regression modeling. A semiquantitative meta-analysis evaluated 38 earlier publications that contain information on height at the onset of diabetes. RESULTS: Children <1 year of age were shorter than their peers by 1 standard deviation, whereas those from 3 years to near puberty were taller by approximately 0.3 standard deviation. Adjusting for parental height caused this difference to disappear for the older children but not for the infants. The meta-analysis results paralleled these observations. CONCLUSIONS: Taller children generally seem to experience increased risk for development of diabetes mellitus type 1, except perhaps during infancy or early adolescence. This observation may have implications regarding pathogenesis of this disorder. PMID- 11875145 TI - Appendicostomy for antegrade enema: effects on somatic and psychosocial functioning in children with myelomeningocele. AB - OBJECTIVE: To assess changes in somatic variables, psychosocial functioning, mental health and self-esteem after appendicostomy for antegrade enema (MACE) in children with myelomeningocele (MMC). METHODS: We performed the MACE procedure in 20 children, aged 6.3 to 17.0 years. Twelve patients had an open and 8 had a laparoscopic operation. Somatic function was assessed preoperatively and 6 and 16 months postoperatively. Psychosocial functioning and mental health were assessed preoperatively and 6 months postoperatively by a structured clinical interview and standardized questionnaires, Youth Self-Report, Child Behavior Checklist, and Harter's Self-Perception Profile for Adolescents. RESULTS: Preoperatively, 9 patients had fecal leaks several times a week, 7 had leaks 1 to 4 times per month, and 4 were continent. The corresponding numbers 6 and 16 months postoperatively were 0, 6, and 14 and 0, 3, and 16. Six patients have had postoperative stoma complications that required surgery; 5 had a well-functioning stoma thereafter, and 1 had a colostomy 7 months later. Preoperatively, 14 patients reported considerable, 4 moderate, 2 minor, and 0 no psychosocial problems related to bowel control and emptying regimen. Six months postoperatively, the corresponding numbers were 1, 2, 6, and 11. The standardized questionnaires revealed substantial psychopathology in the MMC patients. The Child Behavior Checklist and Youth Self-Report scores were not significantly changed postoperatively, but global self-esteem score and close friends score from the Self-Perception Profile for Adolescents were significantly improved. CONCLUSIONS: Fecal incontinence and constipation were greatly reduced by MACE. Children with MMC often have psychological and psychosocial problems, but important improvements in self-esteem and psychosocial function were observed 6 months after the MACE procedure. PMID- 11875147 TI - Seven-year follow-up of vaccine response in extremely premature infants. AB - OBJECTIVE: To assess the immune response of 7-year-old former extremely preterm (PT) infants to routine childhood immunizations. METHODS: Sixteen PT (<29 weeks and <1000 g) infants, followed since their primary immunizations at the recommended chronological ages, and 16 age-matched full-term (FT) control subjects were evaluated at 7 years of age. Antibodies to Haemophilus influenzae type b polyribosylribitol phosphate (Hib-PRP), tetanus, pertussis, diphtheria, polio, and hepatitis B (HBsAb) were measured. RESULTS: The FT group had higher antidiphtheria geometric mean titers (GMT) than the PT group (1.07 vs 0.36 IU/mL). All FT and 13 of 16 PT had protective diphtheria antibody titers (>0.1 IU/mL). The tetanus GMT were 4.22 IU/mL (FT) and 1.99 IU/mL (PT). All children had protective tetanus titers (>0.01 IU/mL). Pertussis titers did not differ between FT and PT. Hib-PRP GMT were higher in FT than in PT (3.21 vs 1.41 microg/mL). All children had anti-PRP > or = 0.15 microg/mL; 12 of 16 FT and 10 of 16 PT had levels > or = 1.0 microg/mL. Polio serotype 1 and 2 GMT were similar between groups, and all children had protective titers (> or = 8). Polio serotype 3 GMT were 59 (FT) and 24 (PT) Karber units; all FT and 12 of 16 PT had protective titers. Among children who had received hepatitis B vaccine, GMT were similar in FT and PT children (120 vs 186 mIU/mL, and similar proportions of children (11 of 16 FT and 12 of 14 PT) had protective HBsAb titers (>10 mIU/mL). CONCLUSIONS: At 7 years of age, PT children had lower antibody titers to many vaccine antigens than FT children. However, most PT children maintained antibody titers in the protective range. PMID- 11875146 TI - A pediatric, practice-based, randomized trial of drinking and smoking prevention and bicycle helmet, gun, and seatbelt safety promotion. AB - OBJECTIVE: To prevent early adolescent health risk behaviors and to maintain or improve safety behaviors, we compared the effects of 2 interventions, delivered through pediatric primary care practices. The interventions, based on an office systems' approach, sought to prevent early drinking and smoking or to influence bicycle helmet use, gun storage, and seatbelt safety for children who were followed from fifth/sixth grades through eighth/ninth grades. DESIGN: Settings and Participants. Twelve pediatric practices in New England were paired according to practice size and assigned randomly within pairs to deliver the multicomponent interventions, which built on pediatric primary care clinicians performing as counselors and role models during health supervision visits and other office encounters. INTERVENTION: One intervention arm focused on alcohol and tobacco use. The other intervention arm focused on gun safety, bicycle helmet, and seatbelt use. Office systems provided infrastructure that supported the clinician's role. Clinician messages encouraged family communication and rule setting about the issues of the middle school years. The intervention was initiated during a health supervision visit and continued for 36 months. Both child and parent received quarterly newsletters to reinforce the clinician messages. OUTCOME MEASURES: The primary outcomes were ever drinking alcohol, ever smoking, ever using smokeless tobacco, using a bicycle helmet in the previous year, using a seatbelt in the previous 30 days, and guns in the child's home in locked storage. RESULTS: The pediatric practices recruited 85% (N = 3525) of the practices' fifth/sixth grade children and their responding parents. We obtained 36 months' follow-up data on 2183 child-parent pairs. Chart audit verified that the intervention was implemented. Additional data from interviews and surveys showed that parents, children, and pediatric clinicians found the intervention useful. Despite this, comparisons between the 2 study arms show no significant intervention effects in the prevention of alcohol and tobacco use or gun storage or seatbelt safety. There was a negative effect in the alcohol arm. Only bicycle helmet use showed a positive outcome. CONCLUSION: With rigorous evaluation, 2 office interventions failed to produce desired outcomes. Coordinated multiple settings for prevention interventions are probably necessary. PMID- 11875148 TI - Practice variation in the emergency management of croup. AB - OBJECTIVE: To explore the effect of physician training background on the emergency management of croup. METHODS: Two community hospitals with a general emergency department (ED) staffed by board-certified emergency medicine (EM) practitioners were the setting for this study. At both sites, pediatricians (PED) or pediatric emergency medicine specialists (PEM) managed acute pediatric visits during evening and weekend hours. Retrospective patient cohorts (6 months to 6 years) with a primary discharge diagnosis of croup were identified from a 1-year period. Data abstraction was performed by a registered nurse who was blinded to the study hypothesis. RESULTS: There were 229, 92, and 209 patients in the PED, PEM, and EM cohorts, respectively, reflecting the practice of 69 physicians (19 PED, 12 PEM, and 38 EM). The groups had similar rates of admission and prescription of steroids at discharge. In regression models that incorporated all recorded clinical variables, EM patients were more likely to have received a chest radiograph (odds ratio [OR]: 6.6; 95% confidence interval [CI]: 3.1--14), racemic epinephrine (OR: 6.5; 95% CI: 3.1--14), albuterol in the ED (OR: 3.0; 95% CI: 1.4--6.4), and parenteral steroids (OR: 3.6; 95% CI: 2.1--6.3) and were less likely to have received oral steroids (OR: 0.41; 95% CI: 0.26--0.64). For the EM cohort, adjusted mean length of ED visit was 40 minutes longer (95% CI: 6.8--72) and mean direct costs were $90 higher (95% CI: $27--$153). Regression models comparing the PEM and PED cohorts revealed no significant management differences. CONCLUSION: Compared with physicians with a pediatric background, rates of resource utilization were higher for EM-trained physicians who managed uncomplicated cases of croup. PMID- 11875149 TI - 2001 Job Lewis Smith award acceptance address. PMID- 11875150 TI - Studies of feeding intolerance in very low birth weight infants: definition and significance. PMID- 11875151 TI - Oxygen and resuscitation: beyond the myth. PMID- 11875152 TI - Science informing policy: The National Institute of Child Health and Human Development's contribution to reading. PMID- 11875153 TI - Two wrongs don't make a right: harm aggravated by inaccurate information on the Internet. AB - There has been much concern expressed in the literature about the use of medical information on the Internet by patients and families. Although much work has been done to quantify the misinformation available on the Internet, there have not been reports of actual harm to children resulting from this misinformation. We present the case of a 1-year-old boy whose clinical course of diarrhea was complicated not only by inaccurate advice given by the emergency room physician, but also by the same advice received from the Internet. PMID- 11875154 TI - Use of photoscreening for children's vision screening. AB - This statement asserts that all children should be screened for risk factors associated with amblyopia. Guidelines are suggested for the use of photoscreening as a technique for the detection of amblyopia and strabismus in children of various age groups. The American Academy of Pediatrics favors additional research of the efficacy and cost-effectiveness of photoscreening as a vision screening tool. PMID- 11875155 TI - Health supervision for children with sickle cell disease. AB - Sickle cell disease (SCD) is a group of complex genetic disorders with multisystem manifestations. This statement provides pediatricians in primary care and subspecialty practice with an overview of the genetics, diagnosis, clinical manifestations, and treatment of SCD. Specialized comprehensive medical care decreases morbidity and mortality during childhood. The provision of comprehensive care is a time-intensive endeavor that includes ongoing patient and family education, periodic comprehensive evaluations and other disease-specific health maintenance services, psychosocial care, and genetic counseling. Timely and appropriate treatment of acute illness is critical, because life-threatening complications develop rapidly. It is essential that every child with SCD receive comprehensive care that is coordinated through a medical home with appropriate expertise. PMID- 11875156 TI - Health care of young children in foster care. AB - Greater numbers of infants and young children with increasingly complicated and serious physical, mental health, and developmental problems are being placed in foster care. All children in foster care need to receive initial health screenings and comprehensive assessments of their medical, mental, dental health, and developmental status. Results of these assessments must be included in the court-approved social services plan and should be linked to the provision of individualized comprehensive care that is continuous and part of a medical home. Pediatricians have an important role in all aspects of the foster care system. PMID- 11875157 TI - Skateboard and scooter injuries. AB - Skateboard-related injuries account for an estimated 50 000 emergency department visits and 1500 hospitalizations among children and adolescents in the United States each year. Nonpowered scooter-related injuries accounted for an estimated 9400 emergency department visits between January and August 2000, and 90% of these patients were children younger than 15 years. Many such injuries can be avoided if children and youth do not ride in traffic, if proper protective gear is worn, and if, in the absence of close adult supervision, skateboards and scooters are not used by children younger than 10 and 8 years, respectively. PMID- 11875158 TI - Guidelines for pediatric cardiovascular centers. AB - Pediatric cardiovascular centers should aim to provide high-quality therapeutic outcomes for infants and children with congenital and acquired heart diseases. This policy statement describes critical elements and organizational features of centers in which high-quality outcomes have the greatest likelihood of occurring. Center elements include noninvasive diagnostic modalities, cardiac catheterization, cardiovascular surgery, and cardiovascular intensive care. These elements should be organizationally united in centers in which pediatric cardiac physician specialists and specialized pediatric staff work together to achieve and surpass existing quality-of-care benchmarks. PMID- 11875159 TI - Selecting and using the most appropriate car safety seats for growing children: guidelines for counseling parents. AB - Despite the existence of laws in all 50 states requiring the use of car safety seats or child restraint devices for young children, more children are still killed as passengers in car crashes than from any other type of injury. Pediatricians and other health care professionals need to provide up-to-date, appropriate information for parents regarding car safety seat choices and proper use. Although the American Academy of Pediatrics is not a testing or standard setting organization, this policy statement discusses the Academy's current recommendations based on the peer-reviewed literature available at the time of publication and sets forth some of the factors that parents should consider before selecting and using a car safety seat. PMID- 11875160 TI - High-frequency oscillatory ventilation: "Please, keep your eyes on me," said the patient. PMID- 11875161 TI - Glutaric aciduria type 1 and nonaccidental head injury. PMID- 11875162 TI - Magnetic resonance imaging and kernicterus. PMID- 11875163 TI - Fever phobia. PMID- 11875164 TI - "Show me the money!". PMID- 11875165 TI - Payment confusion. PMID- 11875166 TI - Sinuses, staph, strep, patients, and families. PMID- 11875167 TI - A double-blind, placebo-controlled study of modified-release methylphenidate in children with attention-deficit/hyperactivity disorder. AB - OBJECTIVE: To compare the efficacy, safety, and tolerability of once-daily administration of modified-release methylphenidate (MPH MR) with placebo in children with attention-deficit/hyperactivity disorder (ADHD). METHODS: The study was a 3-week, double-blind, 32-site, randomized clinical trial comparing MPH MR with placebo. Children were 6 to 16 years of age, had a diagnosis of ADHD, and had not failed a previous trial of stimulant treatment for ADHD. After a 1-week, single-blind, placebo-washout period, participants received a once-daily dose of MPH MR or placebo, which was started with 1 capsule (20 mg) and individually titrated up to a maximum of 3 capsules (60 mg). The primary outcome measure was specified as a reduction in ADHD symptom severity from the teacher version of the 10-item Conners' Global Index. Investigators, teachers, and parents evaluated safety. RESULTS: The study randomized 321 children: 158 to MPH MR and 163 to placebo. Children in the MPH MR group were started on a dose of 20 mg/d and reached a mean dose of 40.7 mg/d (1.28 mg/kg/d) at endpoint. Compared with placebo, MPH MR significantly reduced ADHD symptoms ratings on the teacher version of the 10-item Conners' Global Index, on the parent version of the Conners' Global Index, on the parent assessment of global efficacy, and on investigator assessment of global improvement. The most common adverse events in the MPH MR group were headache, anorexia, abdominal pain, and insomnia. Only anorexia occurred at a rate that was significantly greater than placebo. CONCLUSION: MPH MR administered once daily in the morning is effective and safe in controlling ADHD symptoms throughout the school day. PMID- 11875168 TI - Systemic activity of inhaled steroids in 1- to 3-year-old children with asthma. AB - OBJECTIVE: To study the systemic activity of inhaled steroids in young children. METHODS: Forty children with mild asthma aged 1 to 3 years were studied in a 3 way crossover, randomized, placebo-controlled, double-blind trial. Treatment with inhaled fluticasone propionate, 200 microg twice daily delivered via pressurized metered-dose inhaler (pMDI) and Babyhaler (FP400), was compared with budesonide, 200 microg twice daily delivered via pMDI and NebuChamber (BUD400), and to placebo. The Babyhaler was primed before use. Knemometry was used to detect systemic steroid activity. It was performed with a hand-held knemometer after 1 and 4 weeks of treatment. The increase in lower-leg length within this 3-week period was used as the outcome measure. The intention-to-treat population was analyzed by analysis of variance. RESULTS: The increases in the lower-leg length during placebo, BUD400, and FP400 treatments were 85, 45, and 34 microm/d, respectively (adjusted mean). The growth in lower-leg length was significantly reduced from both steroid treatments. The difference between BUD400 and placebo was -40 microm/d (n = 25; 95% confidence interval [CI]: -8 to -72). The difference between FP400 and placebo was -51 microm/d (n = 26; 95% CI: -19 to 83). The difference between FP and BUD was -11 microm/d and was not statistically significant (n = 28; 95% CI: 20 to -42). CONCLUSION: FP and BUD are both systemically active in children 1 to 3 years old when administered for 4 weeks from their dedicated spacer devices in daily doses of 400 microg with no difference between the 2 steroid regimens. These findings call for studies of clinical side effects from these treatments of preschool children. PMID- 11875169 TI - Outpatient and hospital visits associated with otitis media among American Indian and Alaska native children younger than 5 years. AB - OBJECTIVE: To describe the burden of otitis media (OM) among American Indian and Alaska Native (AI/AN) children. METHODS: OM morbidity among AI/AN younger than 5 years was evaluated using OM-associated outpatient visit and hospitalization rates. These rates were compared with outpatient and hospitalization rates for the general US population of children younger than 5 years. AI/AN children who were younger than 5 years and receiving care through the Indian Health Service or tribally operated facilities and US children younger than 5 years of age were studied. RESULTS: From 1994--1996, the average annual rate of AI/AN OM-associated outpatient visits was 138 per 100 children younger than 5 years. Among AI/AN children younger than 1 year (infants), these rates were almost 3 times greater than those for US infants (318 vs 110 visits per 100 infants, respectively). AI/AN children 1 to 4 years of age had rates 1.5 times greater than US children of the same age (107 vs 65 visits per 100 children, respectively). AI/AN children also experienced higher rates of OM-associated hospitalization than did US children (5643 vs 2440 per 100 000 infants, 823 vs 665 per 100 000 1- to 4-year olds). CONCLUSION: We found that AI/AN children, especially AI/AN infants, have higher OM-associated outpatient and hospitalization rates than those for the general US population of children. The disparity in rates suggests that additional prevention programs and continued resources are needed to reduce OM morbidity among AI/AN children. PMID- 11875170 TI - Nutritional factors that affect the postnatal metabolic adaptation of full-term small- and large-for-gestational-age infants. AB - OBJECTIVE: To document metabolic adaptation to ex utero life in small- (SGA) and large-for-gestational-age (LGA) infants in relation to fetal nutrition and postnatal feeding practices. METHODS: In a prospective study, 65 SGA (< or = second centile) and 39 LGA (> or = 98th centile) full-term infants were recruited. Anthropometry was performed within the first 48 hours. There was full support of breastfeeding and close clinical observation. Blood glucose and ketone body (kb) concentrations were measured prefeed for the first 7 postnatal days. Infants were exclusively breastfed (BF), breastfed with formula milk supplementation (FS), or exclusively formula milk fed (FF). RESULTS: Within the SGA group, a measure of "thinness," the midarm circumference/head circumference ratio, was significantly correlated to the number of episodes of blood glucose < 2.00 mmol/L. Epoch (age at sampling) analysis in this group showed no difference in blood glucose levels across the different feeding groups but revealed a statistically significant greater kb concentration for infants who were exclusively breastfed. For SGA infants, the median peak kb concentration (peak kb) was significantly different for BF, FS, and FF groups. Multiple regression analysis for the SGA group demonstrated that peak kb concentration was negatively related to the volume of formula milk, independent of blood glucose levels and neonatal anthropometry. For LGA infants, low blood glucose levels were offset by kb concentrations equivalent to those observed in infants who were appropriate for gestational age. CONCLUSION: Neonatal ability to generate kb when blood glucose values are low depends more on successful breastfeeding than on size for gestational age or neonatal nutritional status. Routine blood glucose monitoring of LGA infants with no additional risk factors is not necessary. Routine formula milk supplementation for LGA and SGA infants should not be recommended. PMID- 11875171 TI - Hypoplasia of medullary arcuate nucleus in unexpected late fetal death (stillborn infants): a pathologic study. AB - OBJECTIVES: To evaluate the frequency, morphology, and pathogenesis (primary or secondary) of the abnormally developed medullary arcuate nucleus (ARCn) in stillbirths. METHODS: We examined 26 stillbirths (24 antepartum, 2 intrapartum) that had a gestational age between 25 and 40 weeks and a normal karyotype. All of the stillborns were described as well-developed, with body length and weight proportional to their gestational age. Each case was submitted to complete autopsy examination, which included a systematic gross and microscopic evaluation of the body, the placental disk, and the umbilical cord and membranes. The brainstem was the particular focus of the histologic examination. The study of the various nuclei (nucleus hypoglossus, dorsal vagus motor nucleus, tractus solitarii nucleus, nucleus ambiguus, trigeminal tractus and nucleus, arcuate nucleus, and ventrolateral reticular formation and its neurons and parabrachial/Kolliker-Fuse complex) was performed on transversal serial sections through the entire pons and medulla oblongata. The histologic analysis was supplemented by volumetric reconstruction and immunohistochemical detection of both apoptosis and proliferating cell nuclear antigen. RESULTS: Histologic examination showed abnormalities of the medulla oblongata ARCn in 9 fetuses (35%). In 8, a marked hypoplasia was evident, characterized by a volume reduction of the nucleus accompanied by neuronal depletion, whereas in 1 fetus the nucleus was completely absent (agenesis). The absence of gliosis, the negativity of the proliferating cell nuclear antigen analysis, and the similarities in apoptotic indices between the hypoplastic and well-developed arcuate are in keeping with a primary developmental defect. This anomaly is frequently associated with hypoplasia of the reticular formation and chronic hypoxia. CONCLUSIONS: A high frequency of hypoplasia of the ARCn occurs in fetuses who have died "sine causa," ie, in a similar manner to that observed in sudden infant death syndrome. Chemoreceptors, although not involved in reflexogenic oxygenation in fetal life, become of vital importance intrapartum and postpartum; therefore, whenever impaired in the course of development, chemoreceptors may underlie cardioventilatory abnormalities critical to sudden infant death syndrome. PMID- 11875172 TI - Increased activity of lysosomal enzymes in the peritoneal fluid of bacterial peritonitis. AB - OBJECTIVE: The activity of lysosomal enzymes is increased in the cerebrospinal fluid of bacterial meningitis, suggesting that inflammation may cause leakage of lysosomal enzymes into the extracellular fluid. Our objective was to study the activity of 3 lysosomal enzymes in cell-free peritoneal fluid of patients with peritoneal inflammation. METHODS: The beta-galactosidase, beta-glucuronidase, and alpha-mannosidase activity (nmol 4-methylumbelliferone/mL/h); the total, polymorphonuclear, and mononuclear cell number; and chemical parameters were determined in the peritoneal fluid of 26 patients with culture-positive acute bacterial peritonitis, 13 patients (under antibiotic treatment) with culture negative bacterial peritonitis, 6 patients with acute mesenteric lymphadenitis, and 26 control subjects who were operated on for surgical conditions without peritoneal inflammation. RESULTS: The median beta-galactosidase activity in the culture-positive bacterial peritonitis, mesenteric lymphadenitis, and controls was 175 (range: 63--2210), 50 (range: 37--56), and 16 (range: 8--32), respectively. The beta-glucuronidase was 488 (range: 79--998), 53 (range: 27- 98), and 15 (range: 3--22), respectively. The alpha-mannosidase was 801 (range: 100-3172), 78 (range: 33--157), and 41 (range: 16--63), respectively. The differences of the enzyme activities among the groups of the subjects studied were significant, with the exception of the alpha-mannosidase activity between mesenteric lymphadenitis and controls. There was no significant correlation between the enzyme activities and the cytologic or chemical parameters studied. CONCLUSIONS: The elevation of the lysosomal enzymes' activity in the peritoneal fluid of patients with bacterial peritonitis seems to be a reliable index of peritoneal infection. Of the enzymes studied, the beta-glucuronidase and beta galactosidase activities provide the best means for diagnosing bacterial inflammation of the peritoneal cavity. PMID- 11875173 TI - Delayed diagnosis of retinoblastoma: analysis of degree, cause, and potential consequences. AB - OBJECTIVE: To assess the degree, cause, and consequence of delays from presenting signs to diagnosis of retinoblastoma. METHODS: A retrospective chart review was conducted of 64 consecutive patients who presented to the Memorial Sloan Kettering Cancer Center with newly diagnosed retinoblastoma. Seven patients with a positive family history were excluded. RESULTS: The median times from presenting signs to diagnosis for patients with unilateral and bilateral disease were 1.5 and 2.25 months (range: 0--46), respectively; for those who presented with leukocoria and strabismus, median times were 1.5 (range: 0--46) and 2.5 months (range: 0--24). Parents noted the first signs in 75% of the cases. Seventy seven percent delayed seeking treatment, and primary care physicians (PCPs) delayed referral in 30%. Only 3 patients were referred from PCPs solely for physical examination findings. No adverse consequence of delayed diagnosis could be established clearly, but a trend toward eye loss being associated with longer delays in patients with bilateral retinoblastoma was noted. CONCLUSION: Leukocoria and strabismus secondary to retinoblastoma are usually first recognized by relatives rather than PCPs. At routine visits, PCPs should inform parents about the importance of reporting eye abnormalities, and children whose parents complain of leukocoria (white, shiny, jello-like eye) should be referred promptly to an ophthalmologist regardless of whether an absent red reflex is appreciated. PMID- 11875174 TI - Maternal influences on 5- to 7-year-old girls' intake of multivitamin-mineral supplements. AB - OBJECTIVE: To examine diet quality of girls who do or do not take multivitamin mineral (MVM) supplements and to evaluate predictors of girls' MVM use, including maternal eating behaviors, MVM use, beliefs, attitudes, and perceptions about child feeding, eating, and health. DESIGN: Participants were 192 mother and daughter pairs. Daughters were categorized as MVM supplement users or nonusers based on whether girls were consistently given MVM supplements at 5 and 7 years. Girls' and mothers' nutrient and food group intakes, maternal child-feeding practices, and maternal eating behavior were compared between the groups. RESULTS: Mothers who used MVM supplements were more likely to give MVM supplements to daughters. Excluding nutrients from MVM supplements, MVM users and nonusers did not differ in vitamin and mineral intake, either for girls or mothers, and patterns of food group intake were similar for users and nonusers. Mothers of MVM users reported the following: higher levels of pressuring their daughters to eat healthier diets, more monitoring of daughters' food intake, more success in dieting for weight control, more positive evaluations of their success in eating healthy diets, and lower body mass indexes than mothers who did not give MVMs to daughters. CONCLUSIONS: Daughters' MVM supplement use was predicted by mothers' beliefs, attitudes, perceptions, and practices regarding mothers' own eating and child feeding practices, rather than by daughters' diet quality. For both MVM users and nonusers, daughters' food group servings were below recommendations, whereas vitamin and mineral intakes exceeded recommendations, a pattern indicative of girls' relatively high intakes of fortified foods. Mothers should be encouraged to foster healthier patterns of food intake in daughters, rather than providing MVM supplements. PMID- 11875175 TI - Adolescent reports of physician counseling for smoking. AB - OBJECTIVE: Physicians can play an important role in reducing adolescent smoking by counseling their adolescent patients. The appropriate delivery of smoking prevention and cessation messages depends on adequate screening of adolescents, identification of smokers, and adolescents' willingness to disclose their smoking. The present study assessed adolescent reports of physician screening and counseling and adolescents' willingness to disclose smoking, as well as demographic and health status differences in these rates. METHODS: Adolescents (n = 5016), ages 16 to 19, completed a survey on smoking and health. Reports of the prevalence of physician screening, counseling, and adolescents' willingness to disclose their smoking were examined, and logistic regression analyses assessed demographic and health status differences in these prevalence estimates. RESULTS: Overall, 43.4% of the sample reported physician screening, 42.1% reported receiving counseling, and only 28.8% of adolescents reported both. Furthermore, 79.3% of smokers reported that they would admit their smoking if asked. Screening, counseling, and disclosure rates differed by gender, neighborhood income level, smoking status, and asthma status. CONCLUSIONS: More intensive provider-delivered intervention is needed. Efforts should focus on helping providers to identify smoking correctly and to communicate appropriate prevention or cessation messages. Persistence and sensitivity with boys, experimental smokers, and youths with chronic health conditions should be a focus of provider training, because the lower willingness of these youths to disclose their smoking may be a barrier to their identification and intervention. PMID- 11875176 TI - Sequential acquisition of toilet-training skills: a descriptive study of gender and age differences in normal children. AB - OBJECTIVE: To compare the ages, by gender, at which normally developing children acquire individual toilet-training skills and to describe the typical sequence by which children achieve complete toileting success. METHODS: A longitudinal survey was conducted of a cohort of children who were 15 to 42 months of age and attending 4 pediatric practices in the Milwaukee area (2 inner city and 2 suburban) from 1995 through 1997. Parents completed background surveys, and each child's development was assessed using the Bayley Scales of Infant Development II. Each week for 12 to 16 months, parents completed a training status survey (TSS). The TSS, designed for this project, included information on daily toilet training status (eg, number of urine successes on a 5-point scale) and 28 weekly toilet-training behaviors rated on a 5-point Likert scale from 1 (never) to 5 (always). Children were considered to have acquired a particular skill when they received a rating of 4 or 5 on the TSS scale. The median age and interquartile range for children for each toilet-training skill are reported for girls and boys separately using survival curve analysis. Ages at which each gender achieved these skills are compared using a log-rank test. RESULTS: The study included 126 girls and 141 boys; 88% were white. Parents submitted a total of 10 741 weekly surveys (range: 1--73; median: 49 per child). Girls demonstrated toilet-training skills at earlier ages than boys. The median ages for "staying dry during the day" were 32.5 months (95% confidence interval: 30.9--33.7) and 35.0 months (95% confidence interval: 33.3--36.7) for girls and boys, respectively. The median ages for readiness skills for girls and boys, respectively, were as follows: "showing an interest in using the potty," 24 and 26 months; "staying dry for 2 hours," 26 and 29 months; "indicating a need to go to the bathroom," 26 and 29 months. There was a marked concordance in the sequences in which girls and boys achieve individual skills. In addition, the interquartile ranges of the toileting skills varied from 6.9 to 11.4 months in girls and from 7.5 to 14.6 months in boys. CONCLUSIONS: In this study population, girls achieve nearly all toilet training skills earlier than boys, including successful completion. Most children do not master the readiness skills until after the second birthday. The range of normalcy for the attainment of individual skills may vary by as much as a year. PMID- 11875177 TI - Poisoning from a dietary supplement administered during hospitalization. AB - Increasing numbers of persons use dietary supplements (DS). Patients who believe in the effectiveness of DS may continue to take them on admission to a health care facility. We present the case of a child who received a DS on a daily basis as an outpatient, continued its use after admission to the hospital, and became poisoned by it during his hospitalization. PMID- 11875178 TI - Type 1 diabetes mellitus and epilepsia partialis continua in a 6-year-old boy with elevated anti-GAD65 antibodies. AB - A 6-year-old boy presented with epilepsia partialis continua 6 months after diagnosis of type 1 diabetes. Anti-glutamic acid decarboxylase 65 antibodies were found in his serum and cerebrospinal fluid. Anti-epileptic agents did not improve his seizures. High-dose steroids, plasmapheresis, and intravenous immunoglobulin resulted in decreased anti-glutamic acid decarboxylase 65 antibody levels and resolution of his seizures. PMID- 11875179 TI - Retropharyngeal cellulitis in a 5-week-old infant. AB - An infant who presents with acute, unexplained crying requires a thorough examination to identify the source of distress. We report the case of a 5-week old infant who had sudden irritability and was found to have retropharyngeal cellulitis caused by group B Streptococcus. PMID- 11875180 TI - Managing anemia in pediatric office practice: Part 1. PMID- 11875181 TI - Clinical evaluation of substance abuse. PMID- 11875182 TI - Visual diagnosis: an adolescent female who has persistent cough. PMID- 11875183 TI - Rubella vaccine. PMID- 11875184 TI - New views of gene expression in the brain focus on "gene expression tomography". PMID- 11875185 TI - Two closely linked interactive blood pressure QTL on rat chromosome 5 defined using congenic Dahl rats. AB - Previously we reported the construction of a congenic strain, S.LEW, spanning a large region of rat chromosome 5. The Lewis (LEW) strain was the donor, and the Dahl salt-sensitive (S) strain was the recipient. The congenic strain included a blood pressure quantitative trait locus (QTL). In the present work, a series of nine congenic substrains were constructed from S.LEW which defined two closely linked blood pressure QTL in the region previously thought to contain only one. LEW low-blood-pressure alleles at both QTL were required for a major effect on blood pressure. Neither LEW allele alone had a significant effect on blood pressure. The two QTL were localized to regions 6.3 and 4.6 cM, and these were 1.0 cM apart. PMID- 11875187 TI - Blood pressure QTLs identified by genome-wide linkage analysis and dependence on associated phenotypes. AB - Understanding genetic factors that contribute to population-wide variation in blood pressure is likely to benefit prevention and treatment of cardiovascular disease. The aim of the Victorian Family Heart Study is to identify genes for cardiovascular risk in 783 volunteer adult families recruited from the general population. In this preliminary study we sought to identify quantitative trait loci (QTLs) using a genome-wide linkage analysis in 274 adult sibling pairs of average age 24 yr selected without respect to blood pressure. We compared multipoint linkage results for carefully measured systolic (SBP) and diastolic (DBP) pressures before and after statistical adjustment for covariation with sex, oral contraception, age, height, and weight. The average BP was 123/67 (SD: 12/11) mmHg in males (n = 283) and 114/64 (SD: 10/9) mmHg in females (n = 265). Nonparametric Z-scores from multipoint GeneHunter II analysis were "suggestive" (3.1 or more) at four QTLs for SBP (chromosomes 1, 4, 16, and X) but at no QTLs for DBP. Most Z-scores were affected little by adjustment for covariates. However, the SBP QTL on chromosome 16 was obvious only for unadjusted pressures. This population-based quantitative trait analysis has identified more QTLs than any of the eight previous genome-wide scans for blood pressure. Considerable discrepancies between different studies may reflect the presence of false positive results or real biological differences between populations. PMID- 11875186 TI - Cation channel regulation by COOH-terminal cytoplasmic tail of polycystin-1: mutational and functional analysis. AB - Polycystin-1 (PKD1) mutations account for approximately 85% of autosomal dominant polycystic kidney disease (ADPKD). We have shown previously that oocyte surface expression of a transmembrane fusion protein encoding part of the cytoplasmic COOH terminus of PKD1 increases activity of a Ca2+-permeable cation channel. We show here that human ADPKD mutations incorporated into this fusion protein attenuated or abolished encoded cation currents. Point mutations and truncations showed that cation current expression requires integrity of a region encompassing the putative coiled coil domain of the PKD1 cytoplasmic tail. Whereas these loss of-function mutants did not exhibit dominant negative phenotypes, coexpression of a fusion protein expressing the interacting COOH-terminal cytoplasmic tail of PKD2 did suppress cation current. Liganding of the ectodomain of the PKD1 fusion protein moderately activated cation current. The divalent cation permeability and pharmacological profile of the current has been extended. Inducible expression of the PKD1 fusion in EcR-293 cells was also associated with activation of cation channels and increased Ca2+ entry. PMID- 11875188 TI - Analysis of isoproterenol-induced changes in parotid gland gene expression. AB - Parotid gland acinar cells undergo marked hypertrophy and hyperplasia upon systemic exposure to the beta-adrenergic agonist, isoproterenol. This glandular enlargement is accompanied by substantial cellular changes including DNA synthesis, an increase in glandular protein synthesis, and differential changes in RNA transcription. To gain a more detailed understanding of the underlying changes induced by isoproterenol, we have examined the parotid gland gene expression profile of mice up to 24 h post-isoproterenol injection using high density oligonucleotide arrays. Depending upon the exposure time, between 22 and 48 of the approximately 6,500 mouse genes and expressed sequence tags (ESTs) analyzed displayed significant changes in expression patterns. Genes that were previously shown to be repressed (alpha-amylase) or activated (proline-rich proteins) following isoproterenol exposure were found to be similarly affected in this experiment, validating this technique. This study demonstrates that the oligonucleotide array technology is a useful tool for examining isoproterenol induced salivary gland gene expression changes. Using this as a starting point, we can begin to dissect the specific pathways involved in mediating isoproterenol action within the parotid gland. PMID- 11875189 TI - Transcriptional and physiological responses of HepG2 cells exposed to diethyl maleate: time course analysis. AB - Expression levels of 767 genes were measured in HepG2 cells at eight time points (0, 0.5, 1, 6, 12, 16, 20, and 24 h) following exposure to the oxidizing agent, diethyl maleate (DEM). DEM treatment caused an immediate and sustained loss of intracellular GSH, with a concomitant increase in GSSG. From 6-12 h after exposure, there was a substantial increase in the percentage of cells undergoing S phase arrest and apoptosis. Expression profiles of approximately 90% of the genes fell into one of five clusters generated using hierarchical-clustering software, indicating the well-ordered nature of the stress response. The directional movement and timing of induction for many genes matched closely the known physiological role of the proteins they encode. Inhibitors of the cell cycle (CDKN1, CDKN4D, ATM) were induced, whereas cyclins [proliferating cell nuclear antigen (PCNA), cyclin A, cyclin D1, cyclin K] were downregulated during the period from 6--20 h. Likewise, pro-apoptotic genes such as the caspases (CASP9, CASP3, CASP2) and apoptotic protease activating factor (APAF) were induced during the same period. Results of this study indicate that there is a good correlation between time-dependent physiological, biochemical, and gene expression data. PMID- 11875190 TI - Transfer of the Rf-1 region from FHH onto the ACI background increases susceptibility to renal impairment. AB - The genetically hypertensive fawn-hooded (FHH/Eur) rat is characterized by the early presence of systolic and glomerular hypertension, progressive proteinuria (UPV), and albuminuria (UAV), and focal glomerulosclerosis, resulting in premature death from renal failure. Previous studies showed that at least five genetic loci (Rf-1 to Rf-5) were linked to the development of renal impairment. Of these five, Rf-1 appears to play a major role. To study the impact of Rf-1 in the absence of the other loci, we transferred the Rf-1 region of chromosome 1, between the markers D1Mit34 and D1Rat156, Rf-1B for short, onto the genomic background of the normotensive August x Copenhagen Irish (ACI) rat. In this congenic strain, named ACI.FHH-D1Mit34/Rat156 or ACI.FHH-Rf1B, we challenged the renal hemodynamic function of these animals by studying the effects of unilateral nephrectomy (UNX) alone, or combined with N(G)-nitro-L-arginine methyl ester (L NAME)-induced hypertension. Following UNX, the congenic strain developed significantly more UPV and UAV than the ACI progenitor. The differences were even more pronounced when UNX was combined with an L-NAME-induced rise in systolic blood pressure to about 150 mmHg, i.e., the level of hypertension present in the parental FHH strain. These findings indicate that the Rf-1B region of the FHH rat contains at least one gene affecting the susceptibility to progressive renal failure, especially in the presence of an increase in blood pressure. PMID- 11875191 TI - Comparison of gene expression profiling during postnatal development of mouse dentate gyrus and cerebellum. AB - Both the dentate gyrus of the hippocampus and the cerebellar cortex consist mainly of granule cells and develop postnatally. The granule cells in both tissues are presumed to be similar. Changes in gene expression were analyzed during the postnatal development of the dentate gyrus. Altogether, expression patterns of 1,937 genes were determined by adaptor-tagged competitive PCR. More than 90% of the genes belong to groups characterized by elevated expression either at earlier or later stages of development. A majority of the genes expressed showed marked changes during the developmental process, but there was little correlation between gene function and expression, unlike that observed during mouse postnatal cerebellar development. Despite anatomical and physiological similarities between these two processes, the gene expression profiles are completely different. PMID- 11875192 TI - Renal medullary genes in salt-sensitive hypertension: a chromosomal substitution and cDNA microarray study. AB - Substitution of chromosome 13 from Brown Norway BN/SsNHsd/Mcw (BN/Mcw) rats into the Dahl salt-sensitive SS/JrHsd/Mcw (SS/Mcw) rats resulted in substantial reduction of blood pressure salt sensitivity in this consomic rat strain designated SSBN13. In the present study, we attempted to identify genes associated with salt-sensitive hypertension by utilizing a custom, known-gene cDNA microarray to compare the mRNA expression profiles in the renal medulla (a tissue playing a pivotal role in long-term blood pressure regulation) of SS/Mcw and SSBN13 rats on either low-salt (0.4% NaCl) or high-salt (4% NaCl, 2 wk) diets. To increase the reliability of microarray data, we designed a four-way comparison experiment incorporating several levels of replication and developed a conservative yet robust data analysis method. Using this approach, from the 1,751 genes examined (representing more than 80% of all currently known rat genes), we identified 80 as being differentially expressed in at least 1 of the 4 comparisons. Substantial agreements were found between the microarray results and the results predicted on the basis of the four-way comparison as well as the results of Northern blots of 20 randomly selected genes. Analysis of the four-way comparison further indicated that approximately 75% of the 80 differentially expressed genes were likely related to salt-sensitive hypertension. Many of these genes had not previously been recognized to be important in hypertension, whereas several genes/pathways known to be involved in hypertension were confirmed. These results should provide an informative source for designing future functional studies in salt-sensitive hypertension. PMID- 11875193 TI - G protein beta 3 polymorphism and hemodynamic and body composition phenotypes in the HERITAGE Family Study. AB - A C825T polymorphism of the G protein beta3 (GNB3) gene has been reported to be associated with hypertension and obesity. We analyzed the associations between the GNB3 C825T polymorphism and hemodynamic and body composition phenotypes in the sedentary state and their responses to endurance training in mainly normotensive white (n = 473) and black (n = 255) men and women. Blood pressure (BP) and heart rate (HR) were measured at rest and during submaximal exercise at constant power output (50 W), and stroke volume and cardiac output were obtained during exercise. Body composition was assessed with underwater weighing. Baseline systolic BP (SBP) at 50 W was slightly higher in the white CC homozygotes (P = 0.036), whereas in blacks the CC genotype was associated with a lower resting HR (P = 0.012). In blacks, the CC homozygotes showed a greater training-induced reduction in HR at 50 W (P = 0.013) and a similar trend was observed also in whites (P = 0.053). Black women carrying the CC genotype showed significantly greater reductions in resting SBP and diastolic BP (DBP) than the TT homozygotes, whereas in black men the changes in resting BP were similar across the genotypes (P < 0.05 for sex-by-GNB3 interactions). The GNB3 genotype was not associated with baseline body composition in blacks or whites. In blacks, the TT genotype was associated with a greater training-induced decrease in fat mass (P = 0.012) and percent body fat (P = 0.006). These data suggest that DNA sequence variation in the GNB3 locus is not a major modifier of endurance training-induced changes in hemodynamic and body composition phenotypes in healthy but previously sedentary subjects. The GNB3 genotype may play a minor role in HR and body fatness regulation in blacks and in responsiveness of resting BP to endurance training in black women. PMID- 11875194 TI - Gene expression tomography. AB - Gene expression tomography, or GET, is a new method to increase the speed of three-dimensional (3-D) gene expression analysis in the brain. The name is evocative of the method's dual foundations in high-throughput gene expression analysis and computerized tomographic image reconstruction, familiar from techniques such as positron emission tomography (PET) and X-ray computerized tomography (CT). In GET, brain slices are taken using a cryostat in conjunction with axial rotation about independent axes to create a series of "views" of the brain. Gene expression information obtained from the axially rotated views can then be used to recreate 3-D gene expression patterns. GET was used to successfully reconstruct images of tyrosine hydroxylase gene expression in the mouse brain, using both RNase protection and real-time quantitative reverse transcription PCR (QRT-PCR). A Monte-Carlo analysis confirmed the good quality of the GET image reconstruction. By speeding acquisition of gene expression patterns, GET may help improve our understanding of the genomics of the brain in both health and disease. PMID- 11875195 TI - Self-organized complexity in the physical, biological, and social sciences. PMID- 11875196 TI - Fractal dynamics in physiology: alterations with disease and aging. AB - According to classical concepts of physiologic control, healthy systems are self regulated to reduce variability and maintain physiologic constancy. Contrary to the predictions of homeostasis, however, the output of a wide variety of systems, such as the normal human heartbeat, fluctuates in a complex manner, even under resting conditions. Scaling techniques adapted from statistical physics reveal the presence of long-range, power-law correlations, as part of multifractal cascades operating over a wide range of time scales. These scaling properties suggest that the nonlinear regulatory systems are operating far from equilibrium, and that maintaining constancy is not the goal of physiologic control. In contrast, for subjects at high risk of sudden death (including those with heart failure), fractal organization, along with certain nonlinear interactions, breaks down. Application of fractal analysis may provide new approaches to assessing cardiac risk and forecasting sudden cardiac death, as well as to monitoring the aging process. Similar approaches show promise in assessing other regulatory systems, such as human gait control in health and disease. Elucidating the fractal and nonlinear mechanisms involved in physiologic control and complex signaling networks is emerging as a major challenge in the postgenomic era. PMID- 11875197 TI - Allometric scaling of metabolic rate from molecules and mitochondria to cells and mammals. AB - The fact that metabolic rate scales as the three-quarter power of body mass (M) in unicellular, as well as multicellular, organisms suggests that the same principles of biological design operate at multiple levels of organization. We use the framework of a general model of fractal-like distribution networks together with data on energy transformation in mammals to analyze and predict allometric scaling of aerobic metabolism over a remarkable 27 orders of magnitude in mass encompassing four levels of organization: individual organisms, single cells, intact mitochondria, and enzyme molecules. We show that, whereas rates of cellular metabolism in vivo scale as M(-1/4), rates for cells in culture converge to a single predicted value for all mammals regardless of size. Furthermore, a single three-quarter power allometric scaling law characterizes the basal metabolic rates of isolated mammalian cells, mitochondria, and molecules of the respiratory complex; this overlaps with and is indistinguishable from the scaling relationship for unicellular organisms. This observation suggests that aerobic energy transformation at all levels of biological organization is limited by the transport of materials through hierarchical fractal-like networks with the properties specified by the model. We show how the mass of the smallest mammal can be calculated (approximately 1 g), and the observed numbers and densities of mitochondria and respiratory complexes in mammalian cells can be understood. Extending theoretical and empirical analyses of scaling to suborganismal levels potentially has important implications for cellular structure and function as well as for the metabolic basis of aging. PMID- 11875198 TI - Proteins: paradigms of complexity. AB - Proteins are the working machines of living systems. Directed by the DNA, of the order of a few hundred building blocks, selected from 20 different amino acids, are covalently linked into a linear polypeptide chain. In the proper environment, the chain folds into the working protein, often a globule of linear dimensions of a few nanometers. The biologist considers proteins units from which living systems are built. Many physical scientists look at them as systems in which the laws of complexity can be studied better than anywhere else. Some of the results of such studies will be sketched. PMID- 11875199 TI - Turbulence in nature and in the laboratory. AB - Fluid turbulence has attracted the attention of physicists, mathematicians, and engineers for over 100 years, yet it remains an unsolved problem. The reasons for the difficulties are outlined and recent advances in describing its small-scale statistical structure are described. Contrary to traditional notions, new experimental evidence indicates that the small scales are anisotropic, reflecting the overall character of the flow. The consequences of this finding with regard to the long-held postulate of the universality of the small-scale turbulence structure are discussed. PMID- 11875200 TI - What might we learn from climate forecasts? AB - Most climate models are large dynamical systems involving a million (or more) variables on big computers. Given that they are nonlinear and not perfect, what can we expect to learn from them about the earth's climate? How can we determine which aspects of their output might be useful and which are noise? And how should we distribute resources between making them "better," estimating variables of true social and economic interest, and quantifying how good they are at the moment? Just as "chaos" prevents accurate weather forecasts, so model error precludes accurate forecasts of the distributions that define climate, yielding uncertainty of the second kind. Can we estimate the uncertainty in our uncertainty estimates? These questions are discussed. Ultimately, all uncertainty is quantified within a given modeling paradigm; our forecasts need never reflect the uncertainty in a physical system. PMID- 11875201 TI - "Waves" vs. "particles" in the atmosphere's phase space: a pathway to long-range forecasting? AB - Thirty years ago, E. N. Lorenz provided some approximate limits to atmospheric predictability. The details---in space and time---of atmospheric flow fields are lost after about 10 days. Certain gross flow features recur, however, after times of the order of 10--50 days, giving hope for their prediction. Over the last two decades, numerous attempts have been made to predict these recurrent features. The attempts have involved, on the one hand, systematic improvements in numerical weather prediction by increasing the spatial resolution and physical faithfulness in the detailed models used for this prediction. On the other hand, theoretical attempts motivated by the same goal have involved the study of the large-scale atmospheric motions' phase space and the inhomogeneities therein. These "coarse graining" studies have addressed observed as well as simulated atmospheric data sets. Two distinct approaches have been used in these studies: the episodic or intermittent and the oscillatory or periodic. The intermittency approach describes multiple-flow (or weather) regimes, their persistence and recurrence, and the Markov chain of transitions among them. The periodicity approach studies intraseasonal oscillations, with periods of 15--70 days, and their predictability. We review these two approaches, "particles" vs. "waves," in the quantum physics analogy alluded to in the title of this article, discuss their complementarity, and outline unsolved problems. PMID- 11875202 TI - Positive feedback, memory, and the predictability of earthquakes. AB - We review the "critical point" concept for large earthquakes and enlarge it in the framework of so-called "finite-time singularities." The singular behavior associated with accelerated seismic release is shown to result from a positive feedback of the seismic activity on its release rate. The most important mechanisms for such positive feedback are presented. We solve analytically a simple model of geometrical positive feedback in which the stress shadow cast by the last large earthquake is progressively fragmented by the increasing tectonic stress. PMID- 11875203 TI - Unified scaling law for earthquakes. AB - We propose and verify a unified scaling law that provides a framework for viewing the probability of the occurrence of earthquakes in a given region and for a given cutoff magnitude. The law shows that earthquakes occur in hierarchical correlated clusters, which overlap with other spatially separated correlated clusters for large enough time periods and areas. For a small enough region and time-scale, only a single correlated group can be sampled. The law links together the Gutenberg--Richter Law, the Omori Law of aftershocks, and the fractal dimensions of the faults. The Omori Law is shown to be the short time limit of general hierarchical phenomenon containing the statistics of both "main shocks" and "aftershocks," indicating that they are created by the same mechanism. PMID- 11875204 TI - Self-organization in leaky threshold systems: the influence of near-mean field dynamics and its implications for earthquakes, neurobiology, and forecasting. AB - Threshold systems are known to be some of the most important nonlinear self organizing systems in nature, including networks of earthquake faults, neural networks, superconductors and semiconductors, and the World Wide Web, as well as political, social, and ecological systems. All of these systems have dynamics that are strongly correlated in space and time, and all typically display a multiplicity of spatial and temporal scales. Here we discuss the physics of self organization in earthquake threshold systems at two distinct scales: (i) The "microscopic" laboratory scale, in which consideration of results from simulations leads to dynamical equations that can be used to derive the results obtained from sliding friction experiments, and (ii) the "macroscopic" earthquake fault-system scale, in which the physics of strongly correlated earthquake fault systems can be understood by using time-dependent state vectors defined in a Hilbert space of eigenstates, similar in many respects to the mathematics of quantum mechanics. In all of these systems, long-range interactions induce the existence of locally ergodic dynamics. The existence of dissipative effects leads to the appearance of a "leaky threshold" dynamics, equivalent to a new scaling field that controls the size of nucleation events relative to the size of background fluctuations. At the macroscopic earthquake fault-system scale, these ideas show considerable promise as a means of forecasting future earthquake activity. PMID- 11875205 TI - Predictability of catastrophic events: material rupture, earthquakes, turbulence, financial crashes, and human birth. AB - We propose that catastrophic events are "outliers" with statistically different properties than the rest of the population and result from mechanisms involving amplifying critical cascades. We describe a unifying approach for modeling and predicting these catastrophic events or "ruptures," that is, sudden transitions from a quiescent state to a crisis. Such ruptures involve interactions between structures at many different scales. Applications and the potential for prediction are discussed in relation to the rupture of composite materials, great earthquakes, turbulence, and abrupt changes of weather regimes, financial crashes, and human parturition (birth). Future improvements will involve combining ideas and tools from statistical physics and artificial/computational intelligence, to identify and classify possible universal structures that occur at different scales, and to develop application-specific methodologies to use these structures for prediction of the "crises" known to arise in each application of interest. We live on a planet and in a society with intermittent dynamics rather than a state of equilibrium, and so there is a growing and urgent need to sensitize students and citizens to the importance and impacts of ruptures in their multiple forms. PMID- 11875206 TI - Self-organization, the cascade model, and natural hazards. AB - We consider the frequency-size statistics of two natural hazards, forest fires and landslides. Both appear to satisfy power-law (fractal) distributions to a good approximation under a wide variety of conditions. Two simple cellular automata models have been proposed as analogs for this observed behavior, the forest fire model for forest fires and the sand pile model for landslides. The behavior of these models can be understood in terms of a self-similar inverse cascade. For the forest fire model the cascade consists of the coalescence of clusters of trees; for the sand pile model the cascade consists of the coalescence of metastable regions. PMID- 11875207 TI - Complexity and robustness. AB - Highly optimized tolerance (HOT) was recently introduced as a conceptual framework to study fundamental aspects of complexity. HOT is motivated primarily by systems from biology and engineering and emphasizes, (i) highly structured, nongeneric, self-dissimilar internal configurations, and (ii) robust yet fragile external behavior. HOT claims these are the most important features of complexity and not accidents of evolution or artifices of engineering design but are inevitably intertwined and mutually reinforcing. In the spirit of this collection, our paper contrasts HOT with alternative perspectives on complexity, drawing on real-world examples and also model systems, particularly those from self-organized criticality. PMID- 11875208 TI - Natural variability of atmospheric temperatures and geomagnetic intensity over a wide range of time scales. AB - The majority of numerical models in climatology and geomagnetism rely on deterministic finite-difference techniques and attempt to include as many empirical constraints on the many processes and boundary conditions applicable to their very complex systems. Despite their sophistication, many of these models are unable to reproduce basic aspects of climatic or geomagnetic dynamics. We show that a simple stochastic model, which treats the flux of heat energy in the atmosphere by convective instabilities with random advection and diffusive mixing, does a remarkable job at matching the observed power spectrum of historical and proxy records for atmospheric temperatures from time scales of one day to one million years (Myr). With this approach distinct changes in the power spectral form can be associated with characteristic time scales of ocean mixing and radiative damping. Similarly, a simple model of the diffusion of magnetic intensity in Earth's core coupled with amplification and destruction of the local intensity can reproduce the observed 1/f noise behavior of Earth's geomagnetic intensity from time scales of 1 (Myr) to 100 yr. In addition, the statistics of the fluctuations in the polarity reversal rate from time scales of 1 Myr to 100 Myr are consistent with the hypothesis that reversals are the result of variations in 1/f noise geomagnetic intensity above a certain threshold, suggesting that reversals may be associated with internal fluctuations rather than changes in mantle thermal or magnetic boundary conditions. PMID- 11875209 TI - Wavelet analysis of shoreline change on the Outer Banks of North Carolina: an example of complexity in the marine sciences. AB - The horizontal, shore-perpendicular change in shoreline position along the Outer Banks of North Carolina is found to be a self-affine signal. We measure shoreline change by determining the horizontal change in position of the 0.8-m contour sampled from shore-perpendicular profiles spaced at 20-m intervals along the coast. The profiles are obtained from two light detection and ranging surveys performed in September 1997 and September 1998. For six selected sections of coast, wavelet analysis of the shoreline change signal indicates the signal is self-affine with a scaling exponent that varies from 1.2 to 2.1. This self-affine behavior indicates that the shoreline change signal is nonstationary with long range persistence. A stochastic diffusion model of sediment transport replicates the observed self-affine behavior observed south of Cape Hatteras (scaling exponent between 1.2 and 1.6) whereas a random walk model replicates the signal observed north of Cape Hatteras (scaling exponent approximately 2.0). Because of the finite nature of the data set, there are limits in space and time to the power law behavior of the system. Characteristics of such systems can be described by upper-truncated power laws, which yield the upper limits of power law behavior. Applying an upper-truncated power law to the data for one section of coast, we find an upper limit of 7 km for the maximum continuous alongshore distance eroding or accreting. For the same section of coast, we find upper limits of 25 m for the maximum shore-perpendicular erosion and 11 m for the maximum shore-perpendicular accretion during the study period. PMID- 11875210 TI - Self-organized complexity in economics and finance. AB - This article discusses some of the similarities between work being done by economists and by physicists seeking to contribute to economics. We also mention some of the differences in the approaches taken and seek to justify these different approaches by developing the argument that by approaching the same problem from different points of view, new results might emerge. In particular, we review two newly discovered scaling results that appear to be universal, in the sense that they hold for widely different economies as well as for different time periods: (i) the fluctuation of price changes of any stock market is characterized by a probability density function, which is a simple power law with exponent -4 extending over 10(2) SDs (a factor of 10(8) on the y axis); this result is analogous to the Gutenberg--Richter power law describing the histogram of earthquakes of a given strength; and (ii) for a wide range of economic organizations, the histogram shows how size of organization is inversely correlated to fluctuations in size with an exponent approximately 0.2. Neither of these two new empirical laws has a firm theoretical foundation. We also discuss results that are reminiscent of phase transitions in spin systems, where the divergent behavior of the response function at the critical point (zero magnetic field) leads to large fluctuations. PMID- 11875211 TI - Random graph models of social networks. AB - We describe some new exactly solvable models of the structure of social networks, based on random graphs with arbitrary degree distributions. We give models both for simple unipartite networks, such as acquaintance networks, and bipartite networks, such as affiliation networks. We compare the predictions of our models to data for a number of real-world social networks and find that in some cases, the models are in remarkable agreement with the data, whereas in others the agreement is poorer, perhaps indicating the presence of additional social structure in the network that is not captured by the random graph. PMID- 11875212 TI - Scaling phenomena in the Internet: critically examining criticality. AB - Recent Internet measurements have found pervasive evidence of some surprising scaling properties. The two we focus on in this paper are self-similar scaling in the burst patterns of Internet traffic and, in some contexts, scale-free structure in the network's interconnection topology. These findings have led to a number of proposed models or "explanations" of such "emergent" phenomena. Many of these explanations invoke concepts such as fractals, chaos, or self-organized criticality, mainly because these concepts are closely associated with scale invariance and power laws. We examine these criticality-based explanations of self-similar scaling behavior---of both traffic flows through the Internet and the Internet's topology---to see whether they indeed explain the observed phenomena. To do so, we bring to bear a simple validation framework that aims at testing whether a proposed model is merely evocative, in that it can reproduce the phenomenon of interest but does not necessarily capture and incorporate the true underlying cause, or indeed explanatory, in that it also captures the causal mechanisms (why and how, in addition to what). We argue that the framework can provide a basis for developing a useful, consistent, and verifiable theory of large networks such as the Internet. Applying the framework, we find that, whereas the proposed criticality-based models are able to produce the observed "emergent" phenomena, they unfortunately fail as sound explanations of why such scaling behavior arises in the Internet. PMID- 11875213 TI - Access to quality care matters. PMID- 11875215 TI - Datapoints: trends in combined pharmacotherapy with stimulants for children. PMID- 11875216 TI - Improving medication compliance of a patient with schizophrenia through collaborative behavioral therapy. AB - Introduction by the column editors: Numerous factors influence a patient's decision to accept or reject prescribed medications, including the patient's personal values, environmental conditions, and the quality of the patient physician relationship (1). Guidelines for evaluating and managing noncompliance with medication regimens by patients with schizophrenia take this multidimensional perspective into account, emphasizing functional assessment of nonadherence behaviors and individualized behavior-change strategies to secure and maintain the patient's cooperation (2). Moreover, a collaborative approach to planning pharmacotherapy is required to ensure medication compliance, with a particular emphasis on linking the positive effects of medications with the patient's personal goals and desires for better functioning and quality of life (3). The following case study illustrates the application of principles for enhancing medication compliance in the treatment of a woman diagnosed as having schizophrenia, paranoid type. Strategies presented by Dr. Heinssen include collaborative treatment contracts, analysis of adherence behaviors, and techniques for boosting medication cues and reinforcers in the patient's home. The therapy described was provided in the Life Skills partial hospitalization and psychiatric rehabilitation program, a multidisciplinary, multilevel outpatient service of the now-closed Chestnut Lodge Hospital in Rockville, Maryland. The program integrated medical, social-learning, and cognitive-behavioral interventions for psychosis within a psychiatric rehabilitation framework. PMID- 11875217 TI - Consumer & family information: workplace violence: fact, fiction, and prevention. PMID- 11875218 TI - Clinical computing: use of personal digital assistants in consultation psychiatry. PMID- 11875219 TI - Practical geriatrics: persistent postoperative cognitive decline in an elderly woman with preexisting neuropathology. PMID- 11875220 TI - Common ground: a framework for selecting core quality measures for mental health and substance abuse care. AB - There is widespread interest throughout the mental health system in routine quality assessment to facilitate quality improvement, oversight, purchasing, and consumer choice. In the absence of agreement on a limited number of meaningful and feasible quality measures, delivery systems, payers, managed care organizations, regulators, and accreditors have each implemented unique measures and specifications. The resulting heterogeneity among measures has increased the burden on providers, limited the comparability of results, and hindered efforts to focus limited resources on further development of the most promising measures. Policy makers have initiated efforts for stakeholders to reach consensus on a core set of measures for common use, but barriers to progress remain, including differences in stakeholder needs and trade-offs between prioritizing desirable attributes of measures and representing the mental health system broadly. The authors present a framework for the selection of a core set of measures, clarify divergent perspectives, and make recommendations for further development of core quality measures for mental health care. PMID- 11875221 TI - The DSM-IV Text Revision: rationale and potential impact on clinical practice. AB - One consequence of the longer interval between major revisions of the DSM(from seven years between DSM-III-R and DSM-IV to more than 15 years between DSM-IV and DSM-V) is that the accompanying descriptive text will become increasingly out of step with the psychiatric database. To remedy this problem, the DSM-IV Text Revision (DSM-IV-TR) was published in July 2000. The main objectives of the revision were to review the DSM-IV text and make changes to reflect information newly available since the close of the initial DSM-IV literature review process in mid-1992; to correct errors and ambiguities that have been identified in DSM IV; and to update the diagnostic codes to reflect changes in the ICD-9-CM coding system the U.S. government uses officially for health care reporting. This paper reviews the rationale for the text revision and describes changes that may have an impact on the day-to-day use of DSM-IV. PMID- 11875222 TI - Service use and health status of persons with severe mental illness in full-risk and no-risk medicaid programs. AB - OBJECTIVE: The service use patterns and health status outcomes of Medicaid recipients with severe mental illness in a system that assigned full financial risk to managed care organizations through capitation and a system that paid for mental health care on a no-risk fee-for-service basis were compared. METHODS: With use of a quasi-experimental design, initial interviews (time 1) and follow up interviews six months later (time 2) were conducted among 92 clients in the full-risk group and 112 clients in the no-risk group. Regression models were used to compare self-reported service use and health status between the two groups. RESULTS: Service use patterns differed between the two groups. When symptom severity at time 1 was controlled for, clients in the full-risk group were more likely to have received case management but less likely to report contact with a psychiatrist or to have received counseling than clients in the no-risk group. When health status at time 1 was controlled for, clients in the full-risk group reported poorer mental health at time 2 than clients in the no-risk group. When physical health status at time 1 was controlled for, clients in the full-risk group reported poorer physical health at time 2 than clients in the no-risk group. CONCLUSIONS: Capitation was associated with lower use of costly services. Clients with serious mental illness in the full-risk managed care system had poorer mental and physical health outcomes than those in the no-risk system. PMID- 11875223 TI - The effect of financial risk arrangements on service access and satisfaction among medicaid beneficiaries. AB - OBJECTIVE: The relationship between financial risk arrangements, access to services, and consumer satisfaction with services was assessed in a sample of Medicaid beneficiaries who were enrolled under three different financial risk arrangements for health care and mental health care. METHODS: A survey was mailed to a stratified random sample of 9,449 recipients of Supplemental Security Income. Respondents reported their health and mental health service needs, service use, and satisfaction with services. Access was measured in terms of service needs that were met. RESULTS: Access to services was related to the type of risk arrangement. Respondents who were enrolled in plans that assumed the risk for the cost of services had poorer access to services than respondents who were enrolled in plans that did not assume the risk for the cost of these services. Satisfaction with medical services was negatively related to the plan's assuming the risk for medical expenditures. CONCLUSIONS: Financial risk arrangements may have important implications for service use patterns among persons who have disabilities. Health and mental health policy makers should carefully consider risk arrangements when designing health plans for vulnerable populations. PMID- 11875224 TI - Determinants of client outcomes in self-help agencies. AB - OBJECTIVE: This study assessed the relationship between the outcomes of clients of client-run self-help agencies and attendance at the agency, satisfaction with the agency, psychological disability, and organizationally mediated empowerment, that is, the provision of opportunities for clients to meaningfully participate in decisions about their care and the care of others in the agency. The outcomes assessed were independent social functioning, assisted social functioning, and personal empowerment. METHODS: A total of 255 long-term users of four self-help agencies were interviewed at baseline and six months later. Univariate descriptive analyses as well as t tests describing changes in outcomes were conducted. The relationship of each of the four determinants to the three outcomes, after controlling for baseline status on the given outcome and other covariates, was assessed with structural modeling using Amos software. RESULTS: On average, personal empowerment among the clients of the self-help agencies increased, independent social functioning remained the same, and assisted social functioning decreased during the six-month follow-up period. Multivariate analyses showed a positive association between organizationally mediated empowerment and all three outcomes. CONCLUSIONS: The significant ingredient promoting positive outcomes for clients of self-help agencies appears to be the provision of opportunities for clients to meaningfully participate in decisions about their care and the care of others in the organization. PMID- 11875225 TI - Adherence to medication regimens and participation in dual-focus self-help groups. AB - OBJECTIVE: The authors examined the associations between attendance at self-help meetings, adherence to psychiatric medication regimens, and mental health outcomes among members of a 12-step self-help organization specifically designed for persons with both chronic mental illness and a substance use disorder. METHODS: A sample of members of Double Trouble in Recovery (DTR) was interviewed at baseline and one year later. Correlates of adherence to psychiatric medication regimens at the follow-up interview were identified for 240 attendees who had received a prescription for a psychiatric medication. RESULTS: Consistent attendance at DTR meetings was associated with better adherence to medication regimens after baseline variables that were independently associated with adherence were controlled for. Three baseline variables were associated with adherence: living in supported housing, having fewer stressful life events, and having a lower severity of psychiatric symptoms. In addition, better adherence was associated with a lower severity of symptoms at one year and no psychiatric hospitalization during the follow-up period. CONCLUSIONS: Treatment programs and clinicians should encourage patients who have both mental illness and a substance use disorder to participate in dual-focus self-help groups that encourage the responsible use of effective psychiatric medication, particularly after discharge to community living. Clinicians also should be sensitive to stressful life events and discuss with patients how such events might affect their motivation or ability to continue taking medication. PMID- 11875226 TI - Lifetime use of mental health and substance abuse treatment services by incarcerated women felons. AB - OBJECTIVE: This study examined the lifetime use of alcohol, drug, and mental health treatment services by recently incarcerated women prison inmates. METHODS: A total of 805 women entering a North Carolina prison for new felony charges in 1991 and 1992 were interviewed in person shortly after incarceration. The women were assessed for psychiatric disorders and lifetime use of substance abuse and mental health treatment services. Rates of service use were analyzed by inmate characteristics and were compared with rates for a sample of women in the community in North Carolina. RESULTS: The majority of women inmates reported a history of using substance abuse services or mental health services or both. Those with psychiatric disorders and prison recidivists were the most likely to have used such services. Rates of service use were substantially higher for the inmates than for the women in the community, even when the effects of having a psychiatric or substance use disorder were controlled for. CONCLUSIONS: Many of the incarcerated women in the study met lifetime criteria for alcohol, drug, and mental health disorders that were significantly related to their use of substance abuse and mental health treatment services. The majority of the inmates met criteria for a current disorder despite past treatment. Further research is needed to help in developing programs to reduce women inmates' alcohol, drug, and mental health problems. PMID- 11875227 TI - A level-of-functioning self-report measure for consumers with severe mental illness. AB - OBJECTIVE: Most scales that measure the ability of consumers with severe mental illness to function in community settings are designed to be completed by case managers or other clinicians. The objective of this study was to develop an instrument that could be completed by consumers. METHODS: The 17-item clinician version of the Multnomah Community Ability Scale (MCAS) was adapted for use as a self-report instrument (MCAS-SR). An initial version of the MCAS-SR was reviewed for appropriateness and clarity by 20 consumers and four peer counselors and then revised. Test-retest reliability was studied with 37 consumers, and construct validity was examined in a correlational study of 288 consumers. Further validation involved correlations among consumer self-reports, case manager ratings, and ratings by research interviewers. RESULTS: The instrument was acceptable to consumers; most consumers (80 percent) could complete it without assistance. The total score test-retest intraclass correlation was.91. Internal consistency was high (Cronbach's alphas greater than.80). The total score on the MCAS-SR was correlated with that on the Brief Symptom Inventory (Spearman's rho= .59) and with that on the Mental Component Scale of the Short Form 12 (Spearman's rho=.61). The Spearman's rho correlation between consumer and case manager total scores was.41 for urban consumers and.21 for rural consumers. The correlation between consumer total scores and research interviewer total scores was.57 in the urban sample and.31 in the rural sample. CONCLUSIONS: The MCAS-SR is a reliable self-report instrument and can be valuable as an outcome measure and in treatment planning. PMID- 11875228 TI - Firearm-related suicide among young african-american males. AB - National trends in firearm-related suicides among African-American and white males in the age groups 15 to 19 years and 20 to 24 years from 1979 to 1997 were examined. The rates and percentages of suicide by firearms increased significantly more among African-American males than among white males. During the 19-year period, firearms accounted for about 70 percent and 64 percent of all suicides among males aged 15 to 19 years and 20 to 24 years, respectively. The results support the Surgeon General's 1999 call for greater awareness of the suicide risk among African-American males. PMID- 11875229 TI - Integrating outcomes research into clinical practice. AB - A cohort of 102 psychiatric outpatients with major depressive disorder were followed up for about two years. Prospective ratings were obtained at each medication management visit with two instruments that measured changes in symptoms. The mean time to recovery was very similar to that reported in the National Institute of Mental Health's Collaborative Depression Study. This finding supports the validity of using brief instruments to measure outcomes in routine clinical practice. PMID- 11875230 TI - Gaps in use of antipsychotics after discharge by first-admission patients with schizophrenia, 1989 to 1996. AB - The authors examined gaps in the use of antipsychotic medications during the one year period after discharge in an epidemiological sample of 189 first-admission patients with schizophrenia between July 1989 and January 1996. Sixty-three percent of the patients had one or more such gaps, and 51 percent had gaps of 30 days or longer, with an average total time off medication of about seven months. Most gaps occurred soon after discharge, and 73 percent were initiated by the patient. These data, which were obtained before the widespread use of atypical antipsychotic agents, provide a benchmark against which to examine the impact of the newer medications on adherence and continuity of treatment in the critical early stages of schizophrenia. PMID- 11875231 TI - A comparison of psychiatrists' clinical-impression-based and social workers' computer-generated GAF scores. AB - The authors studied the utility of the DSM-IV Global Assessment of Functioning (GAF) scale for improving interdisciplinary communication about patient care. Discharge GAF scores for 165 discharged inpatients were computer generated by 13 trained unit social workers and derived by eight psychiatrists on the basis of their clinical impressions. Differences between the scores obtained by the two disciplinary groups were tested by using the paired t test and the nonparametric signed-rank test. Agreement between scores for various GAF categories was tested with kappa agreement indexes. Interdisciplinary agreement on discharge GAF scores was observed across diagnostic categories and across most categories of length of stay. The results suggest that social workers, after receiving systematic training in computer-based GAF reports, can provide reasonable assessments of clients' functioning. PMID- 11875232 TI - Treatment histories of patients with a syndrome putatively prodromal to schizophrenia. AB - Recent reports suggest that a symptomatic state that is often prodromal to schizophrenia can be identified prospectively. We examined treatment-seeking histories and psychiatric services received among patients with a syndrome similar to a prodromal state. The records of 47 patients who met the criteria for a prodromal state were reviewed. Most patients had previously sought and received psychiatric services (90 percent), including medications (64 percent), and 51 percent had previously received a psychiatric diagnosis. These data suggest that patients who present with a syndrome putatively prodromal to schizophrenia constitute a clinical population. Research into treatment interventions is indicated. PMID- 11875233 TI - Not a joking matter. PMID- 11875234 TI - Fraudulent claims of combat status in the VA? PMID- 11875235 TI - Caregivers' roles in India. PMID- 11875237 TI - Health information on the Internet. PMID- 11875238 TI - Risperidone-induced ejaculatory disturbances. PMID- 11875239 TI - Clozapine and borderline personality disorder. PMID- 11875241 TI - Screening for bipolar disorder. PMID- 11875242 TI - "Summus atque felicissimus salium": the medical relevance of the liquor alkahest. AB - This paper analyzes the development of the concept of alkahest from its origins in the Paracelsian corpus to its mature form in the works of Joan Baptista van Helmont (1579-1644) and his successors. Historians of science have usually focused on the chemical aspects of the alkahest, taking into account especially the claims that it was a substance capable of dissolving all kinds of matter. This paper shows the medical implications of the alkahest: it was not only a "solvent," but an important means of revealing nature's secrets and of producing medicines. The properties ascribed to the alkahest fit perfectly within Helmontian theories about matter, disease, and cure. PMID- 11875243 TI - Diagnosis and authority in the early-twentieth-century medical practice of Richard C. Cabot. AB - This paper examines diagnostic practices using the early twentieth-century medical literature and the patient correspondence and records from the clinic of Richard Cabot. What shaped medicine's rapidly growing persuasive authority in the twentieth century? Diagnostic expertise demonstrated the doctor's control over disease but offered a service of ambiguous value to patients. Cabot and his peers offered differing views on how new diagnostic techniques would influence their relationships to their patients. In his busy private clinic Cabot put into effect an exacting diagnostic process, modeled on his innovative Clinicopathological Conferences. The people who came to the clinic often sought his technical expertise but accepted his diagnostic practices and opinions sometimes only provisionally. PMID- 11875244 TI - Competent professionals and modern methods: state medicine in British Columbia during the 1930s. AB - Little has been written about the formation of state medicine in early-twentieth century Canada, particularly during the Depression era. Indeed, many historians and policy analysts have assumed that this was a time of stagnation and retrenchment in state health provision. To foster a more nuanced analysis of the formation of the Canadian medical state during the Depression decade, this article focuses on British Columbia and the public health initiatives brought in by the provincial Liberal government of T. D. Pattullo. In B.C., an energetic cadre of policymakers and bureaucrats sought to reform existing services by using professionally educated personnel, centralized administrative hierarchies, community education, and the surveillance of target health populations. Funding from the provincial government and the Rockefeller Foundation permitted considerable expansion in a range of public health sectors that included vital statistics, rural health centers, tuberculosis and venereal disease treatment schemes, and laboratory services. This article tells the story of this important period by bringing together details of the professional and personal lives of key individuals--the majority of whom were men--and exploring the new provincial health programs that were developed in B.C. during the interwar years. PMID- 11875245 TI - "Doctor, are you trying to kill me?": ambivalence about the patient package insert for estrogen. AB - In 1976, the U.S. Food and Drug Administration proposed new requirements for patient labeling for estrogens prescribed for menopausal and postmenopausal women. This paper explores the variety of responses to this proposal from women and their husbands, feminist and consumer activists, physicians, pharmacists, and pharmaceutical manufacturers, as represented in letters written to the FDA. The drug industry and the medical profession opposed patient labeling on the grounds of cost and a resentment of governmental intrusion. Feminists and consumer advocates were in favor of the idea, but the response from current estrogen users was mixed: most women wished to be better informed, but many expressed concern that estrogen would be removed from the market. This ambivalence suggests unresolved tensions regarding conceptions of female aging, the medical management of menopause and aging, informed consent in medicine, and governmental regulation of medical practice. The debate thus represents an important moment in the history of women's health care. PMID- 11875246 TI - Updating the accounts: global mortality of the 1918-1920 "Spanish" influenza pandemic. AB - The influenza pandemic of 1918-20 is recognized as having generally taken place in three waves, starting in the northern spring and summer of 1918. This pattern of three waves, however, was not universal: in some locations influenza seems to have persisted into or returned in 1920. The recorded statistics of influenza morbidity and mortality are likely to be a significant understatement. Limitations of these data can include nonregistration, missing records, misdiagnosis, and nonmedical certification, and may also vary greatly between locations. Further research has seen the consistent upward revision of the estimated global mortality of the pandemic, which a 1920s calculation put in the vicinity of 21.5 million. A 1991 paper revised the mortality as being in the range 24.7-39.3 million. This paper suggests that it was of the order of 50 million. However, it must be acknowledged that even this vast figure may be substantially lower than the real toll, perhaps as much as 100 percent understated. PMID- 11875248 TI - Outcome and quality of life in chronic pancreatitis. PMID- 11875249 TI - "A" is for amylin and amyloid in type 2 diabetes mellitus. AB - Amyloid deposits within the islet of the pancreas have been known for a century. In 1987, the islet amyloid precursor polypeptide (IAPP) amylin (a 37 amino acid) was discovered. Recently there has been an explosion of amylin's importance in the development of type 2 diabetes mellitus (T2DM). This review is intended to share what is understood about amylin derived amyloid and the role it plays in T2DM. Whether islet amyloid is an epiphenomenona, a tombstone, or a trigger it leaves an indelible footprint in greater that 70% of the patients with T2DM. There is current data supporting the damaging role of intermediate sized toxic amyloid particles to the beta cell resulting in a beta cell defect which contributes to a relative deficiency or loss of insulin secretion. Within the islet there is an intense redox stress which may be associated with the unfolding of amylin's native secondary structure compounding its amyloidogenic properties. In addition to the beta cell defect there may be an absorptive defect as a result of amyloid deposition in the basement membranes which form an envelope around the inta-islet capillary endothelium. We have an opportunity to change our current treatment modalities with newer medications and we should attempt to diagnose T2DM earlier and use these newer treatment strategies in combination to decrease glucotoxicity without elevating endogenous insulin and amylin. In the 21st century our goal should be to prevent remodeling, save the pancreatic islet, conquer islet amyloid, and amyloid diabetes. PMID- 11875250 TI - Effect of treatment with different doses of 17-beta-estradiol on insulin receptor substrate-1. AB - CONTEXT: Ovarian hormones modulate insulin sensitivity, but their exact role remains unclear. OBJECTIVE: We tried to determine whether different doses of 17 beta-estradiol cause changes in the regulation of insulin receptor substrate (IRS 1) levels, and if so, the possible implications in insulin sensitivity. DESIGN: Ovariectomized rats were treated with different doses of 17-beta-estradiol at 6, 11 and 16 days. MAIN OUTCOME MEASURES: Immunoprecipitation and Western blotting for IRS-1 were performed in different tissues. RESULTS: We found that estradiol treatment has an influence on the amount of IRS-1 but that it acts in different ways depending on the tissue studied, on the length of treatment, and on the doses employed. CONCLUSIONS: Our results suggest that low concentrations of 17 beta-estradiol could be responsible for the upregulation of insulin receptor substrate 1, increasing insulin sensitivity in muscle and adipose tissue. However, insulin receptor substrate 1 is downregulated with high concentrations of 17-beta-estradiol, thus these high hormone plasma levels could favour insulin resistance in peripheral tissues. The role of 17-beta-estradiol seems to modulate insulin receptor substrate 1 levels in insulin dependent tissues, but in a different manner in each tissue. These novel findings are important for improving knowledge about the possible risk for insulin resistance in women taking oral contraceptives or receiving hormone replacement therapy at menopause. PMID- 11875251 TI - Proceedings of the International Symposium on HCO3- and Cystic Fibrosis. San Diego, California, USA. March 3-5, 2001. PMID- 11875252 TI - Bicarbonate-regulated soluble adenylyl cyclase. AB - Soluble adenylyl cyclase (sAC) represents a novel form of mammalian adenylyl cyclase structurally, molecularly, and biochemically distinct from the G protein regulated, transmembrane adenylyl cyclases (tmACs). sAC possesses no transmembrane domains and is insensitive to classic modulators of tmACs, such as heterotrimeric G proteins and P site ligands. Thus, sAC defines an independently regulated cAMP signaling system within mammalian cells. sAC is directly stimulated by bicarbonate ion both in vivo in heterologously expressing cells and in vitro using purified protein. sAC appears to be the predominant form of adenylyl cyclase (AC) in mammalian sperm, and its direct activation by bicarbonate provides a mechanism for generating the cAMP required to complete the bicarbonate-induced processes necessary for fertilization, including hyperactivated motility, capacitation, and the acrosome reaction. Immunolocalization studies reveal sAC is also abundantly expressed in other tissues which respond to bicarbonate or carbon dioxide levels suggesting it may function as a general bicarbonate/CO(2) sensor throughout the body. PMID- 11875253 TI - The cellular physiology of carbonic anhydrases. AB - Carbonic anhydrases are zinc metalloenzymes that catalyze the reversible hydration of CO(2) to form HCO(3)(-) and protons according to the following reaction: CO(2) + H(2)O <=> H(2)CO(3) <=> HCO(3)(-) + H(+). The first reaction is catalyzed by carbonic anhydrase and the second reaction occurs instantaneously. The carbonic anhydrase (CA) gene family includes ten enzymatically active members, which are major players in many physiological processes, including renal and male reproductive tract acidification, bone resorption, respiration, gluconeogenesis, signal transduction, and formation of gastric acid. The newly identified CA IX (previously called MN) and CA XII are related to cell proliferation and oncogenesis. Carbonic anhydrase isozymes have different kinetic properties and they are present in various tissues and in various cell compartments. CA I, II, III and VII are cytoplasmic, CA V is mitochondrial, and CA VI is present in salivary secretions. CA IV, IX, XII and XIV are membrane proteins: CA IV is a glycosyl-phosphatidylinositol-anchored protein, and CA IX, XII and XIV are transmembrane proteins. The present work will focus on the roles of CA II and CA IV in transepithelial proton secretion and bicarbonate reabsorption processes. The localization of these isoforms in selected epithelia that are involved in net acid/base transport, such as kidney proximal tubules and collecting ducts, and tubules from the male reproductive tract will be reviewed. PMID- 11875254 TI - Carbonic anhydrase: in the driver's seat for bicarbonate transport. AB - Carbonic anhydrases are a widely expressed family of enzymes that catalyze the reversible reaction: CO(2) + H(2)O <=> HCO(3)(-) + H(+). These enzymes therefore both produce HCO(3)(-) for transport across membranes and consume HCO(3)(-) that has been transported across membranes. Thus these enzymes could be expected to have a key role in driving the transport of HCO(3)(-) across cells and epithelial layers. Plasma membrane anion exchange proteins (AE) transport chloride and bicarbonate across most mammalian membranes in a one-for-one exchange reaction and act as a model for our understanding of HCO(3)(-) transport processes. Recently it was shown that AE1, found in erythrocytes and kidney, binds carbonic anhydrase II (CAII) via the cytosolic C-terminal tail of AE1. To examine the physiological consequences of the interaction between CAII and AE1, we characterized Cl(-)/HCO(3)(-) exchange activity in transfected HEK293 cells. Treatment of AE1-transfected cells with acetazolamide, a CAII inhibitor, almost fully inhibited anion exchange activity, indicating that endogenous CAII activity is essential for transport. Further experiments to examine the role of the AE1/CAII interaction will include measurements of the transport activity of AE1 following mutation of the CAII binding site. In a second approach a functionally inactive CA mutant, V143Y, will be co-expressed with AE1 in HEK293 cells. Since over expression of V143Y CAII would displace endogenous wild-type CAII from AE1, a loss of transport activity would be observed if binding to the AE1 C-terminus is required for transport. PMID- 11875255 TI - Regulation of Na+-independent Cl-/HCO3- exchangers by pH. AB - Among human bicarbonate transporters, two major gene families encode Na independent Cl(-)/HCO(3)(-) exchangers: the SLC4 anion exchanger (AE) family, and the SLC26 "sulfate permease" anion transporter family. The SLC4 AE family contains at least three genes, and comprises a subfamily within the larger and phylogenetically more ancient bicarbonate transporter superfamily that includes the Na bicarbonate cotransporters (NBC) and the Na-driven Cl/base exchangers. Mutations in the human AE1 gene cause autosomal dominant spherocytic anemia and distal renal tubular acidosis of both dominant and recessive forms. Anemia is also associated with AE1 mutations in mouse, cow, and zebrafish. Naturally occurring mutations in the human AE2 and AE3 genes have not been detected. The SLC26 family in humans consists of at least 10 members, and includes anion exchangers which exchange chloride for bicarbonate, hydroxyl, sulfate, formate, iodide, and/or oxalate. Mutations in three of these genes cause hereditary disease, including chondrodysplasia (SLC26A2, DTD), diarrhea (A3, down-regulated in adenoma/chloride-losing diarrhea protein: DRA/CLD), and goiter/deafness syndrome (A4, pendrin). Little is known about the acute regulation of these modulators of intracellular and compartmental pH and volume. PMID- 11875256 TI - Sodium-coupled bicarbonate transporters. AB - Together, the Na(+)-coupled HCO(3)(-) transporters and the AE family of anion exchangers (i.e., Cl-HCO3 exchangers) comprise the bicarbonate transporter (BT) superfamily. Virtually all BTs are important for the regulation of intracellular pH (pH(i)) in cells throughout the body. Specific BTs also play roles in cell volume regulation, as well as for the transport of salt and/or acid-base equivalents across many epithelia. Electrogenic Na/HCO3 cotransporters (NBCe's) play key roles in HCO(3)(-) reabsorption by the renal proximal tubule, and HCO(3)(-) secretion by the pancreatic duct. Electroneutral NBC's (NBCn's) regulate pH(i) in vascular smooth muscle and are present in/near axons in the brain. Finally, the Na(+)-driven Cl-HCO3 exchanger (NDCBE's) appear to be the major pH(i) regulators in CNS neurons. A characteristic of most, but not all, BT's is that they are inhibited rather effectively by 4,4'-diisothiocyanostilbene 4,4'-disulfonate (DIDS). PMID- 11875257 TI - The electrogenic Na+/HCO3- cotransporter, NBC. AB - Electrogenic Na(+)/HCO(3)(-) (NBC) function has been characterized in many mammalian tissues including, kidney, pancreas, and brain. Cloning efforts identified a single cDNA, NBC/NBC1, that possesses all the functional attributes of the electrogenic Na(+)/HCO(3)(-) cotransporter. This NBC clone is related to the anion exchangers and thus forms a bicarbonate transporter superfamily. Presently two N-terminal and one C-terminal isoforms are known. All three isoforms appear to arise from the same gene and seem to have identical function. NBC antibodies have localized NBC isoforms in kidney, pancreas, brain, small intestine, colon, epididymis, eye, heart, liver, salivary glands, stomach, and testis. Functionally, NBC appears HCO(3)(-) and Na(+) selective. NBC stoichiometry in Xenopus oocytes is 1 Na(+) : 2 HCO(3)(-), implicating a possible accessory protein interaction. PMID- 11875258 TI - Pancreatic ductal bicarbonate secretion: past, present and future. AB - The pancreatic duct epithelium in the guinea-pig and many other species secretes HCO(3)(-) at concentrations approaching 150 mM. This cannot be explained by conventional models based upon HCO(3)(-) secretion via an anion exchanger at the luminal membrane because: 1) under these conditions, the Cl(-) and HCO(3)(-) concentration gradients would favour HCO(3)(-) reabsorption rather than secretion, and 2) the luminal anion exchanger appears to be inhibited by luminal HCO(3)(-) concentrations of 125 mM or more. There may, however, be a sufficiently large electrochemical gradient to drive HCO(3)(-) secretion across the luminal membrane via an anion conductance. In contrast to earlier studies on rat ducts, the membrane potential E(m) in guinea-pig duct cells does not depolarise appreciably upon stimulation with secretagogues but remains constant at about -60 mV. Consequently, even with 125 mM or more HCO(3)(-) in the lumen and an estimated 20 mM in the cytoplasm, the electrochemical gradient for HCO(3)(-) will still favour secretion to the lumen. Under the same conditions, the intracellular Cl(-) concentration drops to very low levels (approximately 7 mM) presumably because, although Cl(-) may leave freely through the cystic fibrosis transmembrane conductance regulator (CFTR) channels in the luminal membrane, there is no major pathway for Cl(-) uptake across the basolateral membrane. Consequently a HCO(3)(-)-rich secretion may arise as a result of the lack of competition from intracellular Cl(-) for efflux via the anion conductances at the luminal membrane. Whether CFTR, or another anion conductance, provides such a pathway for HCO(3)(-) remains to be seen. PMID- 11875259 TI - 150 mM HCO3(-)--how does the pancreas do it? Clues from computer modelling of the duct cell. AB - Cystic fibrosis (CF) takes its name from the pathological changes that occur in the pancreas. Cystic fibrosis transmembrane conductance regulator (CFTR) is highly expressed in the pancreatic ductal epithelium and plays a key role in ductal HCO(3)(-) secretion. In humans, the pancreatic duct secretes near isotonic NaHCO(3). Experimental data suggests that HCO(3)(-) secretion occurs via apical Cl(-)/HCO(3)(-) exchangers working in parallel with Cl(-) channels (CFTR and calcium activated chloride channels, CaCC). Programming the currently available experimental data into our computer model (based on network thermodynamics) shows that while the anion exchanger/Cl(-) channel mechanism will produce a relatively large volume of a HCO(3)(-)-rich fluid, it can only raise the luminal HCO(3)(-) concentration up to about 70 mM. To achieve secretion of about 150 mM NaHCO(3) it is necessary to modulate the properties of the apical membrane transporters as the secreted fluid flows down the ductal system. On the basis of our computer simulations, we propose that HCO(3)(-) secretion occurs mainly via the exchanger in duct segments near the acini (luminal HCO(3)(-) concentration up to about 70 mM), but mainly via channels further down the ductal tree (raising luminal HCO(3)(-) to about 150 mM). We speculate that the switch between these two secretory mechanisms is controlled by a series of luminal signals (e.g. pH, HCO(3)(-) concentration) acting on the apical membrane transporters in the duct cell. PMID- 11875260 TI - Coordination of pancreatic HCO3- secretion by protein-protein interaction between membrane transporters. AB - Increasing evidence suggests that protein-protein interaction is essential in many biological processes including epithelial transport. In this report, we discuss the significance of protein interactions to HCO(3)(-) secretion in pancreatic duct cells. In pancreatic ducts HCO(3)(-) secretion is mediated by cystic fibrosis transmembrane conductance regulator (CFTR) activated luminal Cl( )/HCO(3)(-) exchange activity and HCO(3)(-) absorption is achieved by Na(+) dependent mechanisms including Na(+)/H(+) exchanger 3 (NHE3). We found biochemical and functional association between CFTR and NHE3. In addition, protein binding through PDZ modules is needed for this regulatory interaction. CFTR affected NHE3 activities in two ways. Acutely, CFTR augmented the cAMP dependent inhibition of NHE3. In a chronic mechanism, CFTR increases the luminal expression of Na(+)/H(+) exchange in pancreatic duct cells. These findings reveal that protein complexes in the plasma membrane of pancreatic duct cells are highly organized for efficient HCO(3)(-) secretion. PMID- 11875261 TI - Functional interactions of HCO3- with cystic fibrosis transmembrane conductance regulator. AB - Disruption of normal cystic fibrosis transmembrane conductance regulator- (CFTR) mediated Cl(-) transport is associated with cystic fibrosis (CF). CFTR is also required for HCO(3)(-) transport in many tissues such as the lungs, gastro intestinal tract, and pancreas, although the exact role CFTR plays is uncertain. Given the importance of CFTR in HCO(3)(-) transport by so many CF-affected organ systems, it is perhaps surprising that relatively little is known about the interactions of HCO(3)(-) ions with CFTR. We have used patch clamp recordings from native pancreatic duct cells to study HCO(3)(-) permeation and interaction with CFTR. Ion selectivity studies shows that CFTR is between 3-5 times more selective for Cl(-) over HCO(3)(-). In addition, extracellular HCO(3)(-) has a novel inhibitory effect on cAMP-stimulated CFTR currents carried by Cl(-). The block by HCO(3)(-) was rapid, relatively independent of voltage and occurred over the physiological range of HCO(3)(-) concentrations. These data show that luminal HCO(3)(-) acts as a potent regulator of CFTR, and suggests that inhibition involves an external anion-binding site on the channel. This work has implications not only for elucidating mechanisms of HCO(3)(-) transport in epithelia, but also for approaches used to treat CF. PMID- 11875262 TI - Selective activation of cystic fibrosis transmembrane conductance regulator Cl- and HCO3- conductances. AB - While cystic fibrosis transmembrane conductance regulator (CFTR) is well known to function as a Cl(-) channel, some mutations in the channel protein causing cystic fibrosis (CF) disrupt another vital physiological function, HCO(3)(-) transport. Pathological implications of derailed HCO(3)(-) transport are clearly demonstrated by the pancreatic destruction that accompany certain mutations in CF. Despite the crucial role of HCO(3)(-) in buffering pH, little is known about the relationship between cause of CF pathology and the molecular defects arising from specific mutations. Using electrophysiological techniques on basolaterally permeabilized preparations of microperfused native sweat ducts, we investigated whether: a) CFTR can act as a HCO(3)(-) conductive channel, b) different conditions for stimulating CFTR can alter its selectivity to HCO(3)(-) and, c) pancreatic insufficiency correlate with HCO(3)(-) conductance in different CFTR mutations. We show that under some conditions stimulating CFTR can conduct HCO(3)(-). HCO(3)(-) conductance in the apical plasma membranes of sweat duct appears to be mediated by CFTR and not by any other Cl(-) channel because HCO(3)( ) conductance is abolished when CFTR is: a) deactivated by removing cAMP and ATP, b) blocked by 1 mM DIDS (4,4'-diisothiocyanostilbene-2,2'-disulfonic acid) in the cytoplasmic bath and, c) absent in the plasma membranes of DeltaF508 CF ducts. Further, the HCO(3)(-)/Cl(-) selectivity of CFTR appears to be dependent on the conditions of stimulating CFTR. That is, CFTR activated by cAMP + ATP appears to conduct both HCO(3)(-) and Cl(-) (with an estimated selectivity ratio of 0.2 to 0.5). However, we found that in the apparent complete absence of cAMP and ATP, cytoplasmic glutamate activates CFTR Cl(-) conductance without any HCO(3)(-) conductance. Glutamate activated CFTR can be induced to conduct HCO(3)(-) by the addition of ATP without cAMP. The non-hydrolysable AMP-PNP (5'-adenylyl imidodiphosphate) cannot substitute for ATP in activating HCO(3)(-) conductance. We also found that a heterozygous R117H/DeltaF508 CFTR sweat duct retained significant HCO(3)(-) conductance while a homozygous DeltaF508 CFTR duct showed virtually no HCO(3)(-) conductance. While we suspect that the conditions described here are not optimal for selectively activating CFTR Cl(-) and HCO(3)( ) conductances, we surmise that CFTR may be subject to dramatic alterations in its conductance, at least to these two anions under distinctly different physiological conditions which require distinctly different physiological functions. That is physiologically, CFTR may exhibit Cl(-) conductance with and/or without HCO(3)(-) conductance. We also surmise that the severity of the pathogenesis in CF is closely related to the phenotypic ability of a mutant CFTR to express a HCO(3)(-) conductance. PMID- 11875263 TI - Microelectrode and impedance analysis of anion secretion in Calu-3 cells. AB - Calu-3 cells secrete HCO(3)(-) in response to cAMP agonists but can be stimulated to secrete Cl(-) with K(+) channel activating agonists. Microelectrode and impedance analysis experiments were performed to obtain a better understanding of the conductances and driving forces involved in these different modes of anion secretion in Calu-3 cells. Microelectrode studies revealed apical and basolateral membrane depolarizations upon the addition of forskolin (V(ap) -52 mV vs. -21 mV; V(bl) -60 mV vs. -44 mV) that paralleled the hyperpolarization of the mucosal negative transepithelial voltage (V(T) -8 mV vs. -23 mV). These changes were accompanied by a decrease in the apical membrane fractional resistance (F(Rap)) from approximately 0.50 to 0.08, consistent with the activation of an apical membrane conductance. The subsequent addition of 1-ethyl-2-benzimidazolinone (1 EBIO), a K(+) channel activator, hyperpolarized V(ap) to -27 mV, V(bl) to -60 mV and V(T) to -33 mV. Impedance analysis revealed the apical membrane resistance (R(ap)) of the forskolin-stimulated cells was less than 20 ohm cm(2), indeed in most monolayers R(ap) fell to less than 5 ohm cm(2). The impedance derived estimate of the basolateral membrane resistance (R(bl)) was approximately 170 ohm cm(2) in forskolin treated cells and fell to 50 ohm cm(2) with the addition of 1 EBIO. Using these values for the R(bl) and the F(Rap) value of 0.08 yields a R(ap) of approximately 14 ohm cm(2) in the presence of forskolin and 4 ohm cm(2) in the presence of forskolin plus 1-EBIO. Thus, by two independent methods, forskolin-stimulated Calu-3 cells are seen to have a very high apical membrane conductance of 50 to 200 mS/cm(2). Therefore, we would assert that even at one tenth the anion selectivity for Cl(-), this high conductance could support the conductive exit of HCO(3)(-) across the apical membrane. We further propose that this high apical membrane conductance serves to clamp the apical membrane potential near the equilibrium potential for Cl(-) and thereby provides the driving force for HCO(3)(-) secretion in forskolin-stimulated Calu-3 cells. The hyperpolarization of V(ap) and V(bl) caused by 1-EBIO provides a driving force for Cl(-) exit across the apical membrane, inhibits the influx of HCO(3)(-) on the Na(+):HCO(3)(-) cotransporter across the basolateral membrane, activates the basolateral membrane Na(+):K:2Cl(-) cotransporter and thereby provides the switch from HCO(3)(-) secretion to Cl(-) secretion. PMID- 11875264 TI - Cystic fibrosis transmembrane conductance regulator and H+ permeability in regulation of Golgi pH. AB - This paper reviews experiments from this lab that have tested the hypothesis that pH of the Golgi (pH(G)) of cystic fibrosis (CF) airway epithelial cells is alkaline compared to normal, that this altered pH affects sialyltransferase and other Golgi enzymes controlling biochemical composition of the plasma membrane and that altered surface biochemistry increases bacterial binding. We generated a plasmid encoding a modified green fluorescence protein-sialyltransferase (GFP-ST) chimera protein that was pH-sensitive and localized to the Golgi when transfected into HeLa cells and also CF and normal or cystic fibrosis transmembrane conductance regulator- (CFTR)-corrected airway epithelial cells. Digital imaging microscopy of these Golgi-localized probes showed that there was no correlation between pH(G) (6.4-7.0) and the presence of CFTR, whether cells were in HCO(3)( )/CO(2)-containing or in HCO(3)(-)/CO(2)-free solutions. Activation of CFTR by raising cell [cAMP] had no effect on pH(G). Thus, CFTR seemed not to be involved in controlling pH(G). Experiments on HeLa cells using an avidin-sialyltransferase chimera in combination with a pH-sensitive fluorescent biotin indicated that even in cells that do not express CFTR, Cl(-) and K(+) conductances of the Golgi and other organelle membranes were large and that pH(G) was controlled solely by the H(+) v-ATPase countered by a H(+) leak. A mathematical model was applied to these and other published data to calculate passive H(+) permeability (P(H+)) of the Golgi, endoplasmic reticulum, trans-Golgi network, recycling endosomes and secrety granules from a variety of cells. An organelle's acidity was inversely correlated to its calculated P(H+). We conclude that the CFTR plays a minor role in organelle pH regulation because other (Cl(-) and K(+)) channels are present in sufficient numbers to shunt voltages generated during H(+) pumping. Acidity of the Golgi (and perhaps other organelles) appears to be determined by the activity of H(+) pumps countered by H(+) leaks. PMID- 11875265 TI - Impaired pancreatic ductal bicarbonate secretion in cystic fibrosis. AB - Patients with cystic fibrosis demonstrate a defect in HCO(3)(-) secretion by their pancreatic duct cells. However, attempts toward understanding or correcting this defect have been hampered by a lack of knowledge regarding the cellular and molecular mechanisms mediating HCO(3)(-) transport in these cells. Recent functional and molecular studies indicate a major role for a basolateral electrogenically-driven Na(+):HCO(3)(-) cotransporter (NBC1) in mediating the transport of HCO(3)(-) into the duct cells. The HCO(3)(-) exits at the lumen predominantly via two recently discovered apical HCO(3)(-) transporters. cAMP, which mediates the stimulatory effect of secretin on pancreatic ductal HCO(3)(-) secretion, potentiates the basolateral Na(+):HCO(3)(-) cotransporter due to generation of a favorable electrogenic gradient as a result of membrane depolarization by Cl(-)-secreting cystic fibrosis transmembrane conductance regulator (CFTR). Two apical HCO(3)(-) transporters drive the secretion of bicarbonate into the pancreatic duct lumen. Molecular and functional studies indicate that CFTR upregulates the expression of these two apical HCO(3)(-) transporters. In addition, CFTR may also upregulate the expression of certain water channels and facilitate the secretion of fluid into the duct lumen. In brief, current research suggests that the defect in pancreatic HCO(3)(-) secretion in patients with cystic fibrosis is multifactorial and involves the alteration in the function/expression of transporters at the basolateral and luminal membrane domains of the duct cells. PMID- 11875266 TI - Cl(-)-dependent HCO3- transport by cystic fibrosis transmembrane conductance regulator. AB - Cystic fibrosis (CF) affects the function of multiple organs. The inability to maintain luminal hydration of ducts leads to their plugging and destruction of the affected organs. An exacerbating problem is the acidic pH of the fluid produced by CF patients' secretory glands. This is best documented for pancreatic secretion. Alkaline fluid secretion requires vectorial transport of electrolytes and of HCO(3)(-). The mechanism of HCO(3)(-) secretion by cystic fibrosis transmembrane conductance regulator (CFTR) expressing cells is not well understood. In the present communication we discuss results suggesting that CFTR itself can transport large amounts of HCO(3)(-) and that HCO(3)(-) transport by CFTR is mediated by a coupled, Cl(-)-dependent process that is different from a simple HCO(3)(-) conductance. PMID- 11875267 TI - Na+/HCO3- cotransport in normal and cystic fibrosis intestine. AB - In a search for the HCO(3)(-) supply mechanisms to the enterocyte we cloned and sequenced an intestinal subtype of the Na(+)HCO(3)(-) cotransporter isoform I (dNBC1), which turned out to be identical to the pancreatic NBC1 subtype (pNBC1). Within the intestine, we found particularly high NBC1 expression levels in the duodenum and proximal colon. Experiments with stripped rabbit duodenum in Ussing chambers revealed that Na(+)HCO(3)(-) cotransport (NBC) and CO(2) hydration/Na(+)/H(+) exchange were equally important duodenal HCO(3)(-) supply pathways and were both upregulated during cAMP-mediated secretion. In the proximal colon, however, HCO(3)(-) secretion was low but NBC1 expression even higher than in the duodenum. Ussing-chamber experiments with an NBC-specific inhibitor revealed that NBC, coupled to basolateral Cl(-)/HCO(3)(-) exchange, was an important alternative Cl(-) supply pathway to Na(+)K(+)2Cl(-) cotransport (NKCC) during cAMP-stimulated colonic Cl(-) secretion. To investigate the functional integrity of anion uptake pathways in the absence of cystic fibrosis transmembrane conductance regulator (CFTR), we fluorometrically assessed NBC and NKCC transport rates and cell volume before and during forskolin-stimulation in isolated colonic crypts from normal and CFTR (-/-) mice. Although forskolin stimulation decreased cell volume only in normal, not in CFTR (-/-) crypts, it activated NBC and NKCC to a similar degree in both normal and CFTR (-/-) crypts. We conclude that, depending on the intestinal segment, NBC1 plays an important role in basolateral HCO(3)(-) or Cl(-) uptake. Expression and activation by cAMP is preserved in CFTR (-/-) intestine. PMID- 11875268 TI - Bicarbonate secretion in the murine gallbladder--lessons for the treatment of cystic fibrosis. AB - The epithelium lining the gallbladder of mammalian species has absorptive and secretory functions. An important function is the secretion of a bicarbonate rich fluid that helps neutralise stomach acid and provides an appropriate environment for intestinal enzymes. In cystic fibrosis (CF) this secretory function is lost. This study concerns the bicarbonate secreting activity of murine gallbladders in vitro using wild type and CF mice and four main questions are considered as follows: a) Does the murine gallbladder secrete bicarbonate electrogenically and is this prevented in CF? b) Can the secretory activity in CF gallbladders be restored by gene therapy or pharmacologically? c) How is the cystic fibrosis transmembrane conductance regulator (CFTR) involved in bicarbonate secretion? d) Does the data offer prospects for the treatment of CF?. Work from both the author's laboratory and the literature will be reviewed. Consideration of the currently available data indicates that the wild type murine gallbladder does secrete bicarbonate electrogenically and that this is absent in CF mice. Further it has been demonstrated that bicarbonate secretory activity can be restored by both gene therapy and by the use of drugs. The role of CFTR in bicarbonate secretion remains equivocal. Much evidence suggests that CFTR can act as a channel for HCO(3)(-) ions as well as Cl(-) ions, while others propose a parallel arrangement of CFTR with a Cl(-)/HCO(3)(-) exchanger is necessary. The matter is further complicated by the regulatory role of CFTR on other transporting activities. Opportunities for possible application to man are discussed. PMID- 11875269 TI - Intestinal bicarbonate secretion in cystic fibrosis mice. AB - Gene-targeted disruption of the cystic fibrosis transmembrane conductance regulator (CFTR) in mice results in an intestinal disease phenotype that is remarkably similar to bowel disease in cystic fibrosis patients. In the intestinal segment downstream from the stomach (i.e., the duodenum), CFTR plays an important role in bicarbonate secretion that protects the epithelium from acidic gastric effluent. In this report, we examine the role of CFTR in cAMP stimulated bicarbonate secretion in the murine duodenum and the mechanisms of acid-base transport that are revealed in CFTR knockout (CF) mice. Ion substitution, channel blocker and pH stat studies comparing duodena from wild type and CF mice indicate that CFTR mediates a HCO(3)(-) conductance across the apical membrane of the epithelium. In the presence of a favorable cell-to-lumen HCO(3)(-) gradient, the CFTR-mediated HCO(3)(-) current accounts for about 80% of stimulated HCO(3)(-) secretion. Exposure of the duodenal mucosa to acidic pH reveals another role of CFTR in facilitating HCO(3)(-) secretion via an electroneutral, 4,4'-diisothiocyanato-stilbene-2,2' disulfonic acid (DIDS) sensitive Cl(-)/HCO(3)(-) exchange process. In CF duodenum, other apical membrane acid-base transporters retain function, thereby affording limited control of transepithelial pH. Activity of a Cl(-)-dependent anion exchanger provides near constant HCO(3)(-) secretion in CF intestine, but under basal conditions the magnitude of secretion is lessened by simultaneous activity of a Na(+)/H(+) exchanger (NHE). During cAMP stimulation of CF duodenum, a small increase in net base secretion is measured but the change results from cAMP inhibition of NHE activity rather than increased HCO(3)(-) secretion. Interestingly, a small inward current that is sensitive to the anion channel blocker, 5-nitro-2(3-phenylpropyl amino)-benzoate (NPPB), is also activated during cAMP stimulation of the CFTR null intestine but the identity of the current is yet to be resolved. Studies to identify the proteins involved in non-CFTR mediated HCO(3)(-) secretion are on going and potentially will provide targets to correct deficient HCO(3)(-) secretion in the CF intestine. PMID- 11875270 TI - Duodenal intracellular bicarbonate and the 'CF paradox'. AB - HCO(3)(-) secretion, which is believed to neutralize acid within the mucus gel, is the most studied duodenal defense mechanism. In general, HCO(3)(-) secretion rate and mucosal injury susceptibility correlate closely. Recent studies suggest that luminal acid can lower intracellular pH (pH(i)) of duodenal epithelial cells and that HCO(3)(-) secretion is unchanged during acid stress. Furthermore, peptic ulcers are rare in cystic fibrosis (CF), although, with impaired HCO(3)(-) secretion, increased ulcer prevalence is predicted, giving rise to the 'CF Paradox'. We thus tested the hypothesis that duodenal epithelial cell protection occurs as the result of pH(i) regulation rather than by neutralization of acid by HCO(3)(-) in the pre-epithelial mucus. Cellular acidification during luminal acid perfusion, and unchanged HCO(3)(-) secretion during acid stress are inconsistent with pre-epithelial acid neutralization by secreted HCO(3)(-). Furthermore, inhibition of HCO(3)(-) secretion by 5-nitro-2-(3-phenylpropylamino) benzoic acid (NPPB) despite preservation of pH(i) and protection from acid-induced injury further question the pre-epithelial acid neutralization hypothesis. This decoupling of HCO(3)(-) secretion and injury susceptibility by NPPB (and possibly by CF) further suggest that cellular buffering, rather than HCO(3)(-) exit into the mucus, is of primary importance for duodenal mucosal protection, and may account for the lack of peptic ulceration in CF patients. PMID- 11875271 TI - pH regulation and bicarbonate transport of isolated porcine submucosal glands. AB - We have previously demonstrated that the airway serous cell line Calu-3 employs a number of pH regulatory mechanisms required for bicarbonate secretion by these cells. The aim of the present study was to investigate the pH regulatory mechanisms of serous cells of freshly isolated submucosal glands (SMG). Porcine SMG were dissected out of pig tracheas obtained from a local slaughterhouse. Single glands were transferred into the chamber of an inverted microscope, immobilized by two holding pipettes and the serous cells loaded with the fluorescent pH probe 2',7'-bis-(2-carboxyethyl)-5,6-carboxyfluorescein (BCECF). Fluorescence was monitored from small areas consisting of up to 20 cells. The fluorescence ratio of the emission after excitation at 488 nm and 436 nm respectively was used to estimate cytosolic pH (pH(i)). Resting pH(i) of SMG cells in the absence of HCO(3)(-)/CO(2) was 7.1 +/- 0.16 (n=24). Addition of a solution buffered with HCO(3)(-)/CO(2) to the bath transiently acidified the cells by 0.18 +/- 0.03 (n=18). pH(i) rapidly recovered to a slightly more alkaline value than baseline pH(i). Removal of the HCO(3)(-)/CO(2) buffer strongly alkalinized SMG cells by 0.2 +/- 0.03 (n=18). To challenge pH regulatory mechanisms we exposed the cells to 20 mmol/L NH4(+) in the absence and presence of HCO(3)(-)/CO(2). In both cases we observed a rapid increase in pH(i) followed by a slight recovery. Washout of NH4(+) strongly acidified the cells. Realkalinization of pH(i) could only be observed in the presence of Na(+). This effect was inhibited by the addition of the specific Na(+)/H(+) exchanger isoform 1 (NHE1) blocker 3-methylsulfonyl-4-piperidinobenzoyl guanidine hydrochloride (HOE 694, 10-100 micromol/L) with an half maximal inhibitory concentration (IC(50)) of approximately 20 micromol/L. Full recovery of pH(i) in the presence of HOE 694 was observed when the cells were bathed in HCO(3)(-)/CO(2) solution. Addition of forskolin (5 micromol/L) in the presence of HCO(3)(-)/CO(2) did not significantly alter pH(i) or change pH(i) recovery after acid loading. We conclude that SMG cells possess both HCO(3)(-) dependent and HCO(3)(-) independent pH(i); regulatory mechanisms that require the presence of extracellular Na(+). Further studies are required to understand whether bicarbonate is only transported to regulate pH(i) or whether this transport determines the overall secretory capacity of SMG serous cells. PMID- 11875272 TI - HCO3- transport in relation to mucus secretion from submucosal glands. AB - The role of HCO(3)(-) transport in relation to fluid secretion by submucosal glands is being studied in sheep, pigs, cats and humans. Optical methods have been developed to measure secretion rates of mucus volume from single glands with sufficient temporal resolution to detect differences in minute-by-minute secretion rates among glands. The ionic composition and viscoelastic properties of the uncontaminated gland mucus are measured with a combination of ratiometric fluorescent indicators, ion-selective microelectrodes, FRAP, and a miniaturized, magnetic force viscometer. Sheep glands secreted basally at low rates, showed small, transient responses to alpha- and beta-adrenergic agonists, and large responses to a cholinergic agonist, carbachol. Peak rates and temporal patterns of responses to carbachol differed markedly among glands. To assess the contribution of HCO(3)(-) transport to gland secretion, we either inhibited Na(+)/K(+)/2Cl(-) cotransporter (NKCC) with bumetanide or replaced HCO(3)(-) with HEPES and gassed with O(2). Bumetanide caused a small, non-significant inhibition of basal secretion, but removal of HCO(3)(-)/CO(2) significantly reduced basal secretion almost by half. Both bumetanide and removal of HCO(3)(-)/CO(2) reduced carbachol-stimulated secretion significantly, with HCO(3)(-) removal having the larger effect: a reduction to 33% of control (P<0.01). The remaining secretory response to carbachol was nearly eliminated by bumetanide. Sheep mucus pH measured with ion selective electrodes was about 0.4 log more acidic than the bath. In humans, we observed the same pattern of responses to agonists and antagonists as in sheep, and observed a mucus pH of 7.0 using 2',7' bis(carboxyethyl)-5,6-carboxyfluorescein (BCECF). We hypothesize that HCO(3)(-) transport is important in the formation of mucus secretion, but that most HCO(3)( ) is scavenged before the final mucus appears at the duct opening. Cystic fibrosis transmembrane conductance regulator's (CFTR) best understood function is as an anion channel, but increasing attention has been given to its role in HCO(3)(-) transport. By analogy with organ-specific CFTR effects on Cl(-) transport, it seems likely that the relative importance of CFTR in HCO(3)(-) transport will also vary across organs. Because lung disease is by far the greatest cause of mortality among people with cystic fibrosis, it is important to determine how loss of CFTR function causes lung disease. We are testing the hypothesis that loss of CFTR alters serous cell secretion in the lungs, and the corollary that such loss contributes to cystic fibrosis (CF) lung disease. CFTR is highly expressed in serous cells of submucosal glands and the Calu-3 serous cell model secretes HCO(3)(-). Human gland serous cells grown in culture and tested for fluid secretion under open circuit conditions showed reduced fluid secretion to all mediators. However, submucosal glands are complex organs containing at least 4 distinct regions and at least that many cell types, making it difficult to predict the consequences on whole-organ function from experiments with individual cell types. Therefore, we have resurrected long-neglected methods for studying whole-gland function, and have attempted to improve them in a variety of ways. We are refining these methods and increasing our understanding of gland function by studying tracheal glands from sheep, pigs and cats. As human tissues become available, they are studied with the best methods presently available. The key questions now being asked are: Is mucus secretion from submucosal glands altered in cystic fibrosis? If so, how is it altered and how does it contribute to CF lung disease? Answering the last question will require an understanding of how glands interact with other regions of the lung. In the context of this meeting, we present preliminary data on the role of HCO(3)(-) in gland mucus secretion. PMID- 11875273 TI - Detection of Cl--HCO3- and Na+-H+ exchangers in human airways epithelium. AB - Molecular species of the Na(+)-H(+) exchanger (NHE) and anion exchanger (AE) gene families and their relative abundance in the human airway regions were assessed utilizing RT-PCR and the RNase protection assay, respectively. Organ donor lung epithelia from various bronchial regions (small, medium, and large bronchi and trachea) were harvested for RNA extraction. Gene-specific primers for the human NHE and AE isoforms were utilized for RT-PCR. Our results demonstrated that NHE1, AE2, and brain AE3 isoforms were expressed in all regions of the human airway, whereas NHE2, NHE3, AE1, and cardiac AE3 were not detected. RNase protection studies for NHE1 and AE2, utilizing glyceraldehyde-3-phosphate dehydrogenase as an internal standard, demonstrated that there were regional differences in the NHE1 mRNA levels in human airways. In contrast, the levels of AE2 mRNA remained unchanged. Differential regional expression of NHE1 isoform may be related to a higher acid load in the tracheal epithelial cells than in epithelia of distal airways. Fluctuations in PCO(2) during inspiration and expiration are probably larger in the tracheal lumen than in the lumen of distal airways with associated larger swings in intracellular pH with each respiratory cycle. Immunohistochemical staining for AE2 protein demonstrated localization to the epithelial cells of human bronchial mucosa. PMID- 11875274 TI - cAMP stimulation of HCO3- secretion across airway epithelia. AB - To test for the presence of HCO(3)(-) transport across airway epithelia, we measured short-circuit current in primary cultures of canine and human airway epithelia bathed in a Cl(-)-free, HCO(3)(-)/CO(2)-buffered solution. cAMP agonists stimulated a secretory current that was likely carried by HCO(3)(-) because it was absent in HCO(3)(-)-free solutions. In addition, the cAMP stimulated current was inhibited by the carbonic anhydrase inhibitor, acetazolamide, and by the apical addition of a blocker of cystic fibrosis transmembrane conductance regulator (CFTR), diphenylamine-2-carboxylate. The current was dependent on Na(+) because it was inhibited by removing Na(+) from the submucosal solution and by inhibition of the Na(+)-K(+)-ATPase with ouabain. The cAMP-stimulated current was absent in cystic fibrosis (CF) airway epithelia. These data suggest that cAMP agonists can stimulate HCO(3)(-) secretion across airway epithelia and that CFTR may provide a conductive pathway for HCO(3)(-) movement across the apical membrane. PMID- 11875275 TI - Regulation and functional significance of airway surface liquid pH. AB - In gastrointestinal tissues, cumulative evidence from both in vivo and in vitro studies suggests a role for the cystic fibrosis transmembrane conductance regulator (CFTR) in apical epithelial bicarbonate conductance. Abnormal lumenal acidification is thus hypothesized to play a role in the genesis of cystic fibrosis (CF) pancreatic disease. However, consensus regarding CFTR's participation in pH regulation of airway surface liquid (ASL) and thus the contribution of ASL pH to the etiology of CF lung disease, is lacking. The absence of data reflects difficulties in both sampling ASL in vivo and modeling ASL biology in vitro. Here we evaluate the evidence in support of a lumenal acidification hypothesis in the CF lung, summarize current knowledge of pH regulation in the normal airway and illustrate how hyper-acidified airway secretions could contribute to the pathogenesis of CF lung disease. PMID- 11875276 TI - PMCA2 mutation causes structural changes in the auditory system in deafwaddler mice. AB - Homozygous deafwaddler mice (dfw/dfw) have a mutation in the gene encoding plasma membrane Ca2+ATPase isoform 2 (Pmca2). They walk with a hesitant and wobbly gait, display head bobbing and are deaf. Light microscopy and transmission electron microscopy were used to evaluate the nature and relationship of morphological changes in the cochlea, spiral ganglion cells and spherical cells of the cochlear nucleus in homozygous and heterozygous mice of different ages and controls. Ultrastructural findings showed that in 7 week old homozygous (dfw) mice, inner hair cells and their afferent terminals were present although outer hair cells appeared apoptotic. Stereocilia were absent from the second and third rows of outer hair cells. Ganglion cells were also present although abnormal in appearance. In older homozygous mutants there was a loss of hair cells and spiral ganglion cells. Remaining ganglion cells in this group contained very few cytoplasmic organelles apart from a few hypertrophied mitochondria. In the anteroventral cochlear nucleus, spherical cell soma size was smaller in all homozygous (dfw) mutants than in heterozygous mice and controls. The ultrastructural appearance of the end bulbs of Held in homozygous mutants was abnormal compared with controls, and in the younger group were seen to be swollen, with less distinct synaptic densities and containing large numbers of small synaptic vesicles arranged in clumps. In the older group these synapses were distorted and contained hypertrophied mitochondria and no synaptic densities could be seen, suggesting that these synapses may be non-functional. This study has shown that in homozygous (dfw) mice structural abnormalities occurred not only in cochlear hair cells but also in the spiral ganglion neurones and spherical cells in the cochlear nucleus. It seems likely that these changes are the result of the Pmca2 mutation and the subsequent accumulation of toxic levels of calcium that may lead to alterations in their functional integrity. PMID- 11875277 TI - Tenascin-R associates extracellularly with parvalbumin immunoreactive neurones but is synthesised by another neuronal population in the adult rat cerebral cortex. AB - The molecular components surrounding a neurone serve as recognition cues for the nerve terminals and glial processes that contact them and the constellations formed by these inputs will therefore be determined by the blend of adhesive and repulsive components therein. Using immunohistochemical methods, we observed that the large extracellular matrix-protein, tenascin-R (Restrictin, J1-160-180, Janusin), associates preferentially with the parvalbumin-positive subpopulation of interneurones within the cerebral cortex. In situ-hybridization indicated that tenascin-R-mRNA was expressed in a subpopulation of nerve cells distinct from that containing parvalbumin, suggesting that this protein's association with the latter is receptor mediated. These nerve cells thus modulate at a distance the composition of the extracellular matrix around parvalbuminneurons. PMID- 11875278 TI - The expression of GAD67 isoforms in zebrafish retinal tissue changes over the light/dark cycle. AB - We show the levels of glutamic acid decarboxylase (GAD), the enzyme catalyzing the conversion of glutamic acid to GABA, changes in zebrafish retinal tissue during the light/dark cycle. Further, we identify two transcripts of the GAD67 gene, full-length GAD67 and the truncated 25 kDa alternative splice variant (ES), as the major GAD isoforms in this tissue. GAD-positive neurons were identified immunocytochemically by probing retinal sections with K2, an antibody to the GAD67 isoform, and with an antibody specific for the 25 kDa splice variant. For both antibodies, GAD-immunoreactivity was observed in horizontal cells in the distal retina and amacrine cells in the proximal retina, with both cell bodies and processes labeled. No apparent difference in K2 labeling pattern was observed in tissue harvested 8 hrs after light offset or onset, whereas ES label was identified in more structures in dark tissue. Quantification of GAD levels was determined by densitometry of Western Blots. The protein content of GAD67 and ES varied between tissue harvested during the light and the dark. ES expression was up-regulated in dark tissue; whereas, full-length GAD67 expression increased in light tissue. In vivo GABA content, measured with high performance liquid chromatography (HPLC), was found to increase in light tissue, paralleling the expression of full-length GAD67 transcripts. Expression of ES did not correlate with measured GABA levels, suggesting this isoform, which lacks the catalytic domain necessary for enzymatic activity, may have a different physiological role in retinal tissue. The inverse expression patterns of full-length GAD67 and ES suggest that alternative splicing of GAD67 may be triggered by the light and/or dark cycle, resulting in a change in inhibitory neurotransmitter content in retinal tissue. PMID- 11875279 TI - Distribution and action of some putative neurotransmitters in the stomatogastric nervous system of the earthworm, Eisenia fetida (Oligochaeta, Annelida). AB - The chemical neuroanatomy of the stomatogastric nervous system of the earthworm, Eisenia fetida, has been investigated, using antibodies raised against serotonin, tyrosine hydroxylase, octopamine, GABA, FMRFamide, proctolin, Eisenia tetradecapeptide and neuropeptide Y. Neurons immunoreactive to these antibodies can be observed in the stomatogastric ganglia. The labelled cells comprise altogether 95.4% of the total number of neurons in the ganglion. Immunoreactive projections were followed between stomatogastric individual ganglia as well as towards the enteric plexus. Intrinsic neurons containing the different signal molecules examined are present along the entire length of the enteric plexus, but serotonin immunoreactive perikarya were only found in the hindgut. The density of the different immunoreactive neurons, except the serotonin ones, is highest in the pharyngeal plexus, and the number of labelled neurons decreases along the alimentary canal towards the hindgut. A number of epithelial cells also reveal tyrosine hydroxylase, octopamine, GABA and Eisenia tetradecapeptide immunoreactivity. The action of some putative neurotransmitters, such as dopamine, octopamine, serotonin and proctolin was tested on foregut preparations. Dopamine and octopamine (10(-6)-10(-4) M) have an excitatory effect on the musculature, whereas the effect of serotonin depends on the actual muscle tension. Following precontraction evoked by acetylcholine, serotonin in low concentrations (10(-7)-10(-6) M) causes relaxation, whereas in higher (10(-4) M) concentration it evokes slight contractions. In preparations at basal tone, serotonin (10(-7)-10(-6) M) evokes contractions of the foregut. Atropine strongly inhibits the action of acetylcholine but is ineffective against serotonin, dopamine and octopamine. Similarly, the Na+ channel blocker tetrodotoxin fails to influence the contractile effect of dopamine, octopamine and serotonin. These results suggest that dopamine, octopamine and serotonin act directly on the muscle cells of the alimentary tract. Proctolin do not evoke any significant effect on the foregut. PMID- 11875280 TI - Sensory neurons influence the expression of cell adhesion factors by cutaneous cells in vitro and in vivo. AB - Dorsal root ganglion (DRG) neurons co-cultured with skin-derived fibroblast-like cells (FLCs) show a strong neurite outgrowth. However, when physical contact between FLCs and neurons is prevented with membrane inserts, the DRG neurons exhibit a low survival and a deficient neurite growth. This indicates that cell adhesion molecules influence neuronal survival and neurite growth in co-cultures. The aim of the present study is to find out if selected adhesion molecules are expressed by cultivated FLCs with and without nervous influences, and/or by normal and denervated whole skin. RT-PCR data show that cultured FLCs and denervated skin express L1, N-CAM, N-cadherin and ninjurin, but not neurofascin or TAG-1. However, cultured FLCs exposed to DRG homogenates and innervated skin express N-cadherin only. Following application of neutralizing L1-, N-cadherin- and ninjurin-antibodies (but not N-CAM-antibodies) in the culture medium the mean number of surviving neurons is decreased. Co-cultures incubated with L1-, N cadherin- or ninjurin-antibodies all show significantly less neurite outgrowth compared to controls. In conclusion, the findings in this paper indicate (i) that FLCs cultured in vitro and denervated whole skin express the cell adhesion factors L1, N-CAM, N-cadherin and ninjurin, (ii) that FLCs treated with neural molecules and innervated whole skin express N-cadherin only, (iii) that L1, N cadherin and ninjurin are important for DRG neurons co-cultured with FLCs in vitro in terms of survival and neurite extension and (iv) that there may exist subpopulations of DRG-neurons with different sensitivities for N-cadherin- and ninjurin-antibodies. PMID- 11875281 TI - Immunocytological localization of the HNK-1 carbohydrate in murine cerebellum, hippocampus and spinal cord using monoclonal antibodies with different epitope specificities. AB - The HNK-1 carbohydrate, an unusual 3'-sulfated glucuronic acid epitope characteristic of many neural recognition molecules, serves as a ligand in neural cell interactions and is differentially expressed in the quadriceps and saphenous branches of the femoral nerve in the PNS of adult mice. Based on these observations, we investigated the possibility that the HNK-1 carbohydrate may be differentially distributed in neurons and fiber tracts also in the CNS thereby contributing to different targeting and guidance mechanisms. We have used antibodies with different HNK-1 epitope specificities to probe for subtle differences in expression patterns. In the adult mouse cerebellum the HNK-1 carbohydrate is detectable in stripe-like compartments in the molecular and Purkinje cell layers, whereas N-CAM and its associated alpha2,8 polysialic acid does not show this compartmentation. In the adult hippocampus, the HNK-1 carbohydrate localizes to perineuronal nets of inhibitory interneurons and marks the inner third of the molecular layer of the dentate gyrus. In the adult spinal cord, HNK-1 labeling is most pronounced in gray matter areas. White matter enriched regions show differential labeling with regard to fiber tracts and antibody specificity. Whereas the different antibodies do not show differences in staining in the cerebellum and the hippocampus, they show differences in staining pattern of fiber tracts and motoneurons in the spinal cord. The HNK-1 expression pattern also differed in the adult spinal cord from that observed at embryonic day 14 and postnatal day 14. Our observations suggest a functional role in the specification of functionally discrete compartments in different areas of the CNS and during development. PMID- 11875282 TI - A light and electron microscopic study of the iron transporter protein DMT-1 in the monkey cerebral neocortex and hippocampus. AB - We have studied by immunocytochemistry, the distribution of DMT-1, a cellular iron transporter responsible for transport of metal irons from the plasma membrane to endosomes, in the normal monkey cerebral neocortex and hippocampus. Light to moderate DMT-1 staining was observed in glial cell bodies in the neocortex, the subcortical white matter, and the hippocampus. Despite light labeling of cell bodies, glial end feet around cortical and subcortical blood vessels were heavily labeled. In the neocortex, the glial cell bodies displayed the morphological features of protoplasmic astrocytes. Labeled glial cells in the subcortical white matter contained dense bundles of glial filaments and were identified as fibrous astrocytes. The observation that DMT-1 was present on astrocytic endfeet suggests that these cells are involved in uptake of iron from endothelial cells. It is possible that the iron could then be redistributed into the extracellular space in the brain parenchyma. PMID- 11875284 TI - Preschool attention deficit hyperactivity disorder: a review of prevalence, diagnosis, neurobiology, and stimulant treatment. AB - The clinical use of stimulant medications for 3- to 6-year-old preschool children who meet diagnostic criteria for attention deficit hyperactivity disorder (ADHD) is becoming more common. A systematic computerized literature search extending back to 1970 identified nine controlled studies of stimulant treatment and two controlled trials of stimulant side effects in preschool ADHD children. Treatment benefits are reported for eight of nine (89%) controlled stimulant trials involving a total of 206 preschool subjects. In comparison with school-aged ADHD youth, there may be a greater variability of stimulant response in ADHD preschoolers. Domains assessing cognition, interpersonal interactions, and hyperactive-impulsive behavior are noted to improve on drugs relative to placebos. Side effects in this age range are generally reported as mild. ADHD preschool children may experience slightly more and different types of stimulant induced side effects compared with older children. High rates of behavior reported as stimulant side effects are found for children receiving a placebo, necessitating a baseline evaluation for medication side effects before stimulants are initiated. Despite the lack of research assessing stimulant effects on the very young and developing brain and the need for more controlled medication trials in this age range, this review of the extant literature finds stimulants to meet evidence based criteria as beneficial and safe for carefully diagnosed ADHD preschool children aged 3 years and older. PMID- 11875285 TI - Lessons from three year olds. PMID- 11875286 TI - In harm's way: toxic threats to child development. AB - Developmental disabilities result from complex interactions of genetic, toxicologic (chemical), and social factors. Among these various causes, toxicologic exposures deserve special scrutiny because they are readily preventable. This article provides an introduction to some of the literature addressing the effects of these toxicologic exposures on the developing brain. This body of research demonstrates cause for serious concern that commonly encountered household and environmental chemicals contribute to developmental disabilities. The developing brain is uniquely susceptible to permanent impairment by exposure to environmental substances during time windows of vulnerability. Lead, mercury, and polychlorinated biphenyls (PCBs) have been extensively studied and found to impair development at levels of exposure currently experienced by significant portions of the general population. High dose exposures to each of these chemicals cause catastrophic developmental effects. More recent research has revealed toxicity at progressively lower exposures, illustrating a "declining threshold of harm" commonly observed with improved understanding of developmental toxicants. For lead, mercury, and PCBs, recent studies reveal that background-population exposures contribute to a wide variety of problems, including impairments in attention, memory, learning, social behavior, and IQ. Unfortunately, for most chemicals there is little data with which to evaluate potential risks to neurodevelopment. Among the 3000 chemicals produced in highest volume (over 1 million lbs/yr), only 12 have been adequately tested for their effects on the developing brain. This is a matter of concern because the fetus and child are exposed to untold numbers, quantities, and combinations of substances whose safety has not been established. Child development can be better protected by more precautionary regulation of household and environmental chemicals. Meanwhile, health care providers and parents can play an important role in reducing exposures to a wide variety of known and suspected neurodevelopmental toxicants that are widely present in consumer products, food, the home, and wider community. PMID- 11875287 TI - Five burning questions. PMID- 11875288 TI - Patterns of psychopathology and dysfunction in clinically referred preschoolers. AB - Despite the growing interest in the use of psychotropic medications in preschoolers, little is known about the clinical presentation of young children referred for psychiatric services. We describe the clinical characteristics, psychiatric disorders, and functioning of preschoolers referred for pediatric psychiatry evaluation. Structured psychiatric interviews assessing lifetime psychopathology by DSM-III-R criteria were completed on clinically referred youth. Family, social, and overall functioning were assessed at intake. From the pool of 1658 consecutive referrals, we identified 200 children less than or equal to ( 3.5 mg/mmol and macroalbuminuria if UACR > 35 mg/mmol. RESULTS: The mean age of the 8029 patients was 66 years, 54% were women. Microalbuminuria was found in 23% and macroalbuminuria in 4% of patients. Microalbuminuria was more prevalent in patients of African American (35%), Hispanic (37%) and Asian (36%) ethnicity, heavy smokers (32%), diabetics (36%) and in patients with ECG left ventricular hypertrophy by both ECG-criteria (29%). Urine albumin/creatinine was positively related to Sokolow-Lyon voltage criteria and Cornell voltage-duration product criteria. In multiple regression analysis, higher UACR was independently associated with older age, diabetes, higher blood pressure, serum creatinine, smoking and left ventricular hypertrophy. Patients smoking > 20 cigarettes/day had a 1.6-fold higher prevalence of microalbuminuria and a 3.7-fold higher prevalence of macroalbuminuria than never-smokers. ECG left ventricular hypertrophy by Cornell voltage-duration product or Sokolow-Lyon criteria was associated with a 1.6-fold increased prevalence of microalbuminuria and a 2.6 fold increase risk of macroalbuminuria compared to no left ventricular hypertrophy on the second ECG. CONCLUSIONS: In patients with moderately severe hypertension, left ventricular hypertrophy on two consecutive ECGs is associated with increased prevalences of micro- and macroalbuminuria compared to patients without persistent ECG left ventricular hypertrophy. High albumin excretion was related to left ventricular hypertrophy independent of age, blood pressure, diabetes, race, serum creatinine or smoking, suggesting parallel cardiac damage and albuminuria. PMID- 11875308 TI - Angiotensin I-converting enzyme gene polymorphism influences chronic hypertensive response in the rat Goldblatt model. AB - BACKGROUND AND OBJECTIVE: In humans, the insertion/deletion polymorphism in the angiotensin (Ang) I converting enzyme (ACE) gene significantly determines ACE activity. The deletion allele induces higher ACE levels and is associated with hypertension in men. In the rat, a microsatellite marker in the ACE gene allows differentiation of the ACE alleles among strains with different ACE levels. We evaluated the effect of genetically determined ACE expression on the development of renovascular hypertension in the rat. METHODS AND RESULTS: Systolic BP (SBP), ACE and angiotensin II (Ang II) levels were measured using the Goldblatt (Gb) model (two kidneys, one clip) in homozygous males of two inbred strains (F2) of Lewis x Brown-Norway (BN) rats. SBP was significantly higher in the BN-Gb rats compared to the Lewis-Gb rats throughout the study (F = 239.6, P < 0.001). An interaction was observed between SBP and strain (F = 2.92, P < 0.01). Plasma ACE activity was 100% higher in the BN-Gb than in the Lewis-Gb rats (P < 0.05). Ang II plasma levels were higher in the BN-sham than in the Lewis-sham rats (255 +/- 22 versus 161 +/- 16 pg/ml, P < 0.05), increased in both Gb groups and correlated significantly with SBP (r = 0.58, P < 0.01). CONCLUSIONS: Genetically determined ACE expression in male rats enhances the chronic hypertensive response after the induction of renovascular hypertension. A relationship between circulating Ang II and the development of hypertension was also observed in this experimental model of genetically modulated hypertension. PMID- 11875309 TI - Nitric-oxide-mediated relaxations in salt-induced hypertension: effect of chronic beta1 -selective receptor blockade. AB - BACKGROUND: Nebivolol is a new beta1-selective adrenergic receptor antagonist with a direct vasorelaxant effect that involves activation of the l-arginine nitric oxide (NO) pathway. Therefore, treatment with nebivolol may protect against endothelial injury in hypertension. OBJECTIVE: To investigate whether chronic selective beta1-blockade with nebivolol could prevent endothelial dysfunction in salt-induced hypertension, and to compare it with atenolol. METHODS: Dahl salt-sensitive rats were treated for 8 weeks with standard chow or chow containing 4% NaCl alone or in combination with nebivolol (10 mg/kg per day) or atenolol (100 mg/kg per day). Isometric tension was continuously recorded in isolated aorta and small mesenteric arteries. Constitutive NO synthase (cNOS) activity was determined by [3H]citrulline assay. RESULTS: Chronic salt administration increased systolic blood pressure by 38 +/- 5 mmHg in salt-treated rats as compared with that in control rats. Both nebivolol and atenolol prevented a salt-induced increase in pressure. cNOS activity was significantly decreased by a high-salt diet. The impairment of endothelium-dependent relaxations in response to acetylcholine in salt-treated rats was prevented only by nebivolol, in both large and small arteries. In contrast, the reduced endothelium-independent relaxations and contractions in response to sodium nitroprusside and endothelin 1, respectively, were restored by both drugs. Nebivolol, but not atenolol, restored cNOS activity. CONCLUSIONS: Despite nebivolol and atenolol having the same blood-pressure-decreasing effect, only nebivolol was able to prevent endothelial dysfunction. This study demonstrates for the first time that the acute NO-mediated vasodilatory action of nebivolol is also present during chronic treatment. Hence, nebivolol might become a new therapeutic tool with which to exert vascular protective effects against end-organ damage in conditions associated with NO deficiency. PMID- 11875310 TI - Simvastatin improves endothelial function in spontaneously hypertensive rats through a superoxide dismutase mediated antioxidant effect. AB - BACKGROUND: Hydroxymethylglutaryl coenzyme A (HMGCoA) reductase inhibitors have beneficial effects beyond their cholesterol-lowering properties. The antioxidant mechanism of HMGCoA reductase inhibitors is not completely understood. OBJECTIVES: To elucidate the antioxidant effect of simvastatin. METHODS: We studied the influence of simvastatin treatment on the development of hypertension, modification of antioxidant systems, and reactivity of aortic rings in Wistar-Kyoto (WKY) and spontaneously hypertensive (SHR) rats. RESULTS: Simvastatin had no effect on blood pressure (BP). Simvastatin treatment (either 1 or 2 mg/kg body weight for 12 or 20 weeks) increased superoxide dismutase (SOD) and glutathione peroxidase (GPx) activities in SHR rats compared with untreated control SHR rats. Carbachol-induced relaxation of aortic rings was impaired in control SHR rats and was restored by simvastatin treatment. Addition of SOD improved the response in control SHR rats and did not have any effect in treated SHR rats. Addition of diethyldithiocarbamic acid, a selective inhibitor of SOD, produced a mild non-significant impairment in carbachol-induced relaxation in control SHR rats, suggesting a deficient antioxidant system in these animals. However, in treated SHR and in WKY rats, impairment of the relaxation was marked, implying that SOD activity in these animals was important to maintain endothelial function. In aortic rings without endothelium from SHR rats, contraction induced by free radicals was substantially higher than in WKY rats. This effect was attenuated in 1-mg-treated rats and abolished in 2-mg-treated rats. CONCLUSIONS: Simvastatin promotes intracellular antioxidant systems, fundamentally SOD, restoring endothelial function but not having any effect on blood pressure. PMID- 11875311 TI - Angiotensin II receptor antagonist improves age-related endothelial dysfunction. AB - BACKGROUND: We previously demonstrated that the angiotensin converting enzyme (ACE) inhibitor, enalapril, prevents the age-related impairment of endothelium dependent hyperpolarization and relaxation mediated by endothelium-derived hyperpolarizing factor (EDHF). OBJECTIVE: To test whether angiotensin II type 1 (AT1) receptor antagonists would also improve age-related endothelial dysfunction. METHODS: Normotensive Wistar-Kyoto (WKY) rats were treated for 3 months with either the AT1 receptor antagonist, candesartan cilexetil (3.5 mg/kg per day; candesartan group), or the ACE inhibitor, enalapril (20 mg/kg per day; enalapril group), from 9 to 12 months of age. Untreated 12-month-old WKY rats (old group) served as controls (n = 7-12). RESULTS: The two treatments decreased systolic blood pressure comparably. EDHF-mediated hyperpolarization in response to acetylcholine (ACh; 10(-5) mol/l) in the presence of norepinephrine in mesenteric arteries was improved in both the candesartan and enalapril groups to a similar extent compared with the old group (candesartan group, -24 +/- 3 mV; enalapril group, -21 +/- 2 mV; old group, -13 +/- 2 mV). EDHF-mediated relaxation was similarly improved in the candesartan and enalapril groups (maximum relaxation: candesartan group, 70 +/- 7%; enalapril group, 63 +/- 8%; old group, 33 +/- 9%). Hyperpolarization and relaxation responses to levcromakalim, an ATP sensitive K+-channel opener, were similar in all groups. CONCLUSIONS: These findings suggest that the AT1 receptor antagonist is as effective as the ACE inhibitor in improving the age-related decline in EDHF-mediated hyperpolarization and relaxation in normotensive rats. Thus AT1 receptor antagonists might serve as novel tools with which to prevent endothelial dysfunction associated with aging. PMID- 11875312 TI - Contribution of sarcoplasmic reticulum Ca2+ to the activation of Ca2+ -activated K+ channels in the resting state of arteries from spontaneously hypertensive rats. AB - OBJECTIVE: Localized release of Ca2+ from the sarcoplasmic reticulum (SR) toward the plasmalemma, sometimes visualized as Ca2+ sparks, can activate Ca2+-activated K+ (KCa) channels. We have already reported that the addition of charybdotoxin (ChTX), a blocker of KCa channels, to the resting state of arteries from spontaneously hypertensive rats (SHR) caused a powerful contraction, suggesting that KCa channels were active in the resting state. This study aimed to determine whether the Ca2+ responsible for activity of KCa channels was derived from SR. METHODS: Possible mechanisms underlying the ChTX-induced contractions were examined in endothelium-denuded strips of femoral, mesenteric, small mesenteric and carotid arteries from 13-week-old SHR and normotensive Wistar-Kyoto (WKY) rats by using selective inhibitors of the Ca2+ spark process. RESULTS: ChTX (100 nmol/l) induced a contraction in the SHR arteries. The ChTX-induced contractions were increased by a moderate membrane depolarization by 15.9 mmol/l K+ and were abolished by nifedipine (100 nmol/l). When SR Ca2+ was depleted by treatment of the strips with ryanodine (10 mumol/l) plus caffeine (20 mmol/l) or with thapsigargin (100 nmol/l), the ChTX-induced contraction was decreased in femoral, mesenteric and small mesenteric arteries and was almost abolished in the carotid artery. A similar phenomenon can be observed in arteries from WKY rats after a moderate membrane depolarization. In both SHR and WKY rats, SR Ca2+-dependent ChTX-induced contraction always represents 20-30% of the maximal K+-induced contraction. CONCLUSIONS: We conclude that activation of KCa channels depended upon influx of Ca2+ through L-type Ca2+ channels and release of Ca2+ from the SR, suggesting that recycling of entering Ca2+ from the superficial SR toward the plasmalemma sufficiently elevated Ca2+ near these channels to activate them. PMID- 11875313 TI - Downregulation of nitric oxide accumulation by cyclooxygenase-2 induction and thromboxane A2 production in interleukin-1beta-stimulated rat aortic smooth muscle cells. AB - BACKGROUND: Cytokines from inflammatory cells do not produce nitric oxide, but stimulate the production of nitric oxide in vascular smooth muscle cells (VSMC). Thromboxane A2 (TXA2) has been believed to have a key role in atherosclerogenesis and post-angioplasty restenosis. OBJECTIVE: To determine whether cytokine-induced nitric oxide production is regulated by the TXA2/prostaglandin H2 (PGH2) receptor. METHODS AND RESULTS: We studied the interleukin-1beta (IL-1beta) induced production of nitric oxide in rat VSMCs using the TXA2/PGH2 receptor antagonists, seratrodast and Bay-u3405, and an agonist, U-46619. Nitrite formation was measured colorimetrically. IL-1beta increased nitrite formation in a time-dependent manner. The nitrite concentration was 1.7 times greater in the presence of seratrodast than that without it. Nitrite accumulation was increased by Bay-u3405, but was decreased in the presence of U-46619, to 44% of that in its absence. Western and Northern blotting showed that seratrodast increased the levels of expression of inducible nitric oxide synthase (iNOS) protein and mRNA in a dose-dependent manner, whereas U-46619 decreased them. We speculated that VSMCs produced TXA2, thereby decreasing nitric oxide production; therefore we measured the accumulation of TXB2 using an enzyme immunoassay. Untreated VSMCs produced about 20 pg/mg protein of TXB2. This was increased by the addition of IL 1beta, to 152.1 +/- 43.0 pg/mg protein after a 24 h incubation; the expression of cyclooxygenase-2 (COX-2) protein was also increased, but there was no effect on the expression of COX-1 and TXA2 synthase. U-63557A, a TXA2 synthase inhibitor, increased the accumulation of nitrite to 1.3-fold that in its absence. CONCLUSIONS: These data suggest that the expression of iNOS and the production of nitric oxide are regulated by the TXA2/PGH2 receptor in IL-1beta-stimulated VSMCs. The endogenous production of TXA2 by the induction of COX-2 from IL-1beta stimulated VSMCs probably downregulated the production of nitric oxide in VSMCs. TXA2/PGH2 receptor inhibitors may contribute to the reduction in formation of atherosclerosis in lesions with vascular injury by enhancing the production of nitric oxide by VSMCs. PMID- 11875314 TI - Reduction of the soluble cyclic GMP vasorelaxing system in the vascular wall of stroke-prone spontaneously hypertensive rats: effect of the alpha1 -receptor blocker doxazosin. AB - OBJECTIVE: The aim of the present study was to analyse the nitric oxide (NO)/cyclic GMP (cGMP) relaxing system in spontaneously hypertensive rats of the stroke-prone substrain (SHRSP). DESIGN: The study was performed in 20-week-old SHRSP rats. A group of normotensive Wistar-Kyoto (WKY) rats was used as control. RESULTS: The endothelium-dependent relaxation to acetylcholine was reduced in SHRSP rats (n = 15). No modifications in the expression of the endothelial nitric oxide synthase were found in the vascular wall of WKY rats (n = 15) and SHRSP rats. SHRSP rats demonstrated an impaired relaxing response to the NO-donor sodium nitroprusside that was accompanied by a reduction in the level of the main second messenger of NO, cyclic GMP. The expression of the soluble guanylate cyclase (sGC) beta1-subunit was markedly reduced in the vascular wall of SHRSP rats. In the experimental model of SHRSP, an increased concentration of catecholamines has been reported. Therefore, we evaluated the effect of an alpha1 receptor blocker, doxazosin, on the NO/cGMP system. Doxazosin [10 mg/kg body weight (bw) per day for 15 days, n = 15] reduced mean arterial pressure (MAP) in SHRSP rats. Treatment with doxazosin preserved the endothelium-independent relaxation response to sodium nitroprusside in aortic segments from SHRSP rats which was associated with an increased expression of the sGC beta1-subunit. A dose of doxazosin (1 mg/kg bw per day, n = 15) that did not modify MAP partially prevented sGC protein expression in the vascular wall. CONCLUSIONS: Independently of the endothelial NO-generating system, impaired vasorelaxation could also result from vascular smooth muscle cell layer dysfunction. Doxazosin treatment improved the endothelial-independent relaxation and preserved the cGMP generating system in the vascular wall of SHRSP rats. PMID- 11875315 TI - Structural factors increase blood pressure through the interaction of resistance vessel geometry with neurohumoral and local factors: estimates in rabbits with renal cellophane-wrap hypertension with intact effectors and during neurohumoral blockade. AB - BACKGROUND: The structural changes in hypertension include narrowing of the lumen of the large resistance vessels and an increase in their wall thickness : lumen ratio. Their haemodynamic role has been controversial. OBJECTIVE: To examine resting haemodynamics and the responses to graded drug-induced changes in tone in renal cellophane-wrap (wrap) and sham-operated (sham) rabbits, when their neurohumoral effectors were intact and during high-level blockade. METHODS: Each rabbit was implanted with a flow probe for measuring cardiac output, had catheters inserted for drug infusions, and underwent mean arterial pressure (MAP) measurements. Resting values were determined and we infused graded doses of dilator and constrictor drugs: acetylcholine or adenosine; angiotensin II or methoxamine. The dilator and constrictor dose-response curves were combined into a single relationship for MAP, cardiac output, total peripheral conductance (TPC) and heart rate; total peripheral resistance (TPR) was estimated as 1/TPC. RESULTS: Throughout the range of vascular tone and with intact effector function, MAP was greater and TPC lower in wrap than in sham rabbits, and cardiac output was the same in both groups. The ratios of wrap : sham slopes of the log dose response regression lines were 0.47 for TPC, 2.04 for TPR and 1.89 for MAP. Thus MAP and TPR responses were enhanced to the same degree in wrap rabbits. During neurohumoral block, baroreflex-mediated heart rate responses were abolished. In addition, resting vascular tone was lower than with intact effectors in both wrap and sham rabbits; however, the ratios of wrap : sham slopes for TPC and TPR were similar to those with intact effectors, whereas the ratio for the slope for MAP was slightly smaller, although still enhanced. CONCLUSION: In wrap hypertension, the enhanced MAP and TPR responses are in accord with an interaction between vascular geometry and the sum of altered neurohumoral + local activity, plus a rarefaction component. PMID- 11875316 TI - Changes in pain perception during treatment with angiotensin converting enzyme inhibitors and angiotensin II type 1 receptor blockade. AB - OBJECTIVES: Besides the well-known role of the angiotensin system in blood pressure control, an interaction of angiotensin and pain perception has been suggested. This study sought to investigate whether an angiotensin converting enzyme inhibitor, which facilitates bradykinins, algesic peptides, and/or an AT1 receptor antagonist may modify hypertension-related hypoalgesia in humans. The study was approved by the ethical committee of our Department. METHODS: A total of 22 hypertensive patients were submitted to dental pulp stimulation to obtain the dental pain threshold and tolerance, and to 24 h blood pressure monitoring together with a control group of 55 normotensives. Then the hypertensives were randomized to enalapril or losartan treatment and were re-evaluated (dental pain perception and ambulatory monitoring) after 8 weeks of the first treatment and after an additional 8 weeks of the second treatment. RESULTS: Untreated hypertensives showed a reduced perception to painful stimuli when compared with normotensives. A significant reduction of both pain threshold and tolerance was observed during the anti-hypertensive treatments (Friedman test: P = 0.007 and P = 0.006, respectively). Pain sensitivity was similar during the two treatments and it did not differ from pain sensitivity values of normotensive controls. ANCOVAs were computed to evaluate the relationship between anti-hypertensive agents and pain sensitivity, after controlling for blood pressure. A 24 h mean pressure served as covariate, removing any effect of blood pressure; a significant difference was observed entering both pain threshold and tolerance as dependent variables (F = 5.28, P = 0.0076; F = 8.16, P = 0.0007, respectively). CONCLUSIONS: Both the angiotensin converting enzyme inhibitor enalapril and the AT1 receptor blocking agent losartan acted similarly on pain threshold and tolerance, pain sensitivity being increased during the two anti-hypertensive treatments. The blood pressure reduction during drug assumption could not account for the pain sensitivity changes observed. The latter may be due to a specific pharmacodynamic mechanism mediated through angiotensin II AT1 receptors. PMID- 11875317 TI - Nitric oxide and potassium channels are involved in brain natriuretic peptide induced vasodilatation in man. AB - OBJECTIVE: Brain natriuretic peptide (BNP) causes vasodilatation but the mechanisms by which this is accomplished are not fully known. The aim of the present study was to determine whether, besides K+Ca2+-channels, nitric oxide (NO) is involved in BNP-induced vasodilatation. METHODS: We studied 10 healthy males twice, in random order, at an interval of 2 weeks. Experiments always started with infusion of BNP (8-16-32-64 pmol/dl per min) into the brachial artery. On the first day this infusion was followed by a second BNP infusion combined with the K+Ca2+-channel-blocker, tetraethylammonium (TEA, 0.1 mg/dl per min), and on the other day by BNP infusion combined with the NO-synthase inhibitor, l-NG-monomethyl arginine (l-NMMA, 0.8 mumol/dl per min). The latter was then followed by a combined infusion of BNP, l-NMMA and TEA. All infusions were separated by a 1 h washout period. Forearm blood flow (FBF) was determined by venous occlusion plethysmography. RESULTS: Mean arterial pressure and heart rate did not change during any of the experiments. BNP alone induced a dose dependent dilatation, which was similar on both days. TEA, l-NMMA, and their combination all reduced the BNP-induced dilatation (P < 0.05). The combined infusion had a significantly greater effect than TEA alone (P = 0.005). BNP infusions were associated with a significant increase in plasma cyclic guanosine monophosphate (cGMP) and C-type natriuretic peptide (CNP) (P < 0.05). CONCLUSIONS: BNP induces arterial vasodilatation not only by opening K+Ca2+ channels, but also via stimulation of NO production. In addition, BNP stimulates net CNP increase. PMID- 11875318 TI - Relationship between muscle sympathetic nerve activity and large artery mechanical vessel wall properties in renal transplant patients. AB - OBJECTIVES: Renal transplant recipients (RTX) show a major impairment of large artery elastic wall properties. Sympathetic overactivity present in patients with renal disease has been shown to alter large artery elasticity; however, in RTX, this issue has not been addressed. The present study therefore investigated a possible relationship between sympathetic activity and large artery distensibility in RTX. METHODS: In 32 patients treated with calcineurin inhibitors (RTX-CI, cyclosporine n = 16, tacrolimus n = 16) mean arterial pressure (MAP, automatic sphygmomanometer), muscle sympathetic nerve activity (MSNA, microneurography) and distensibility coefficients of the brachial and carotid arteries (pulsed Doppler) were measured. Sixteen healthy volunteers (CTR), six patients with calcinneurin inhibitor-free immunosuppression (RTX-AZA) and 12 transplant patients after native kidney nephrectomy (RTX-NC) served as control groups. RESULTS: RTX-CI significantly increased MSNA compared to CTR (36 +/- 3 versus 16 +/- 2 bursts/min, P < 0.05, mean +/- SEM). Both brachial and carotid artery distensibility were decreased in RTX-CI compared to CTR (7 +/- 1 versus 13 +/- 1 +/- 10(-3) /kPa and 17 +/- 1 versus 25 +/- 2 x 10(-3) /kPa, respectively, both P < 0.05). In RTX-CI, a significant inverse correlation between brachial, but not carotid artery distensibility and MSNA (r = -0.46, P < 0.01, r = -0.12, not significant, respectively) was found. Correlation between brachial artery distensibility and MSNA remained statistically significant on separate analysis of cyclosporine- or tacrolimus-treated RTX and after correction for arterial diameter, blood pressure, graft function, age and sex by stepwise multiple regression analysis. Results in RTX-AZA were similar to those in RTX-CI. In contrast, in RTX-NC with MSNA not significantly different from CON (16.6 +/- 2.0 bursts/min), brachial artery distensibility was significantly higher compared to RTX-CI and RTX-AZA (14.2 +/- 2.0 x 10(-3) /kPa, P < 0.05, respectively). CONCLUSIONS: Increased sympathetic nerve activity in renal transplant patients is related to decreased distensibility of the muscular type brachial artery, but not the elastic type carotid artery. PMID- 11875319 TI - Relation of insulin resistance to blood pressure in childhood. AB - OBJECTIVE: To determine the relation between blood pressure and insulin resistance in children and the differences between fasting insulin and the insulin clamp in that relation. METHODS: Children were randomly selected after blood pressure screening of 12 043 fifth--eigth grade Minneapolis, Minnesota, students, with stratification by systolic blood pressure (SBP) percentile [half from the upper 25th percentile (high blood pressure) and half from the lower 75th percentile (low blood pressure)]. Euglycemic insulin clamps were performed with an insulin infusion rate of 1 mU/kg per min and a variable infusion of 20% glucose to maintain plasma glucose at 5.6 mmol/l (100 mg/dl). Insulin sensitivity (Mlbm) is defined as the amount of glucose required to maintain euglycemia (mg glucose infused/kg lean body mass (LBM)/min. RESULTS: Diastolic blood pressure was not significantly correlated with any of the body measurements or laboratory data. SBP was significantly correlated with virtually all measures of body size in males and females. SBP and Mlbm were not significantly correlated in either sex. The correlation between SBP and fasting insulin was significant for boys and girls, but became non-significant after adjustment for BMI. All measures of body fatness were significantly greater in the high blood pressure group, and a significant clustering effect for fasting insulin, BMI, triglycerides, and HDL-C was related to blood pressure. The clustering effect was similar when Mlbm was substituted for fasting insulin and was similar for boys, girls, blacks and whites. CONCLUSIONS: These results suggest that level of blood pressure in children is mediated through body fat and that insulin resistance, as determined by the insulin clamp, does not play a primary role at this age of development. Nevertheless, the clustering effect of the risk factors according to SBP grouping is consistent with an early relation of blood pressure to the insulin resistance syndrome. PMID- 11875320 TI - Silent cerebral white matter lesions in middle-aged essential hypertensive patients. AB - OBJECTIVE: Age, hypertension, diabetes mellitus and a history of cardiovascular disease are the most important factors related to the presence of cerebral white matter lesions (WML), which are a common finding in elderly people. This study investigates which factors related to hypertension per se are associated with the presence of WML in asymptomatic, middle-aged, never-treated essential hypertensive patients. METHODS: A total of 66 untreated essential hypertensive patients of both genders, aged 50-60 years, with neither diabetes mellitus nor evidence of cardiovascular disease, were studied. Hypertensive patients were classified into two groups according to the presence or absence of WML in brain magnetic resonance imaging (MRI). RESULTS: A total of 39 (59.1%) hypertensives showed no WML in brain MRI, and 27 (40.9%) exhibited the presence of WML. Compared with hypertensives without WML, patients with WML showed significantly higher values of both office and 24 h ambulatory blood pressure monitoring (ABPM) systolic, diastolic, mean and pulse pressure. No differences were observed in either the nocturnal fall of blood pressure, or in blood pressure variability, assessed by 24 h standard deviation, among hypertensives with WML. In contrast, the nocturnal decline of heart rate was significantly blunted in patients with WML, compared with those without WML. CONCLUSIONS: Cerebral white matter lesions are a common finding in asymptomatic middle-aged essential hypertensives. The severity of blood pressure elevation seems to be the most important factor related to the presence of WML. Neither the circadian rhythm nor the long-term variability of blood pressure were related to WML. PMID- 11875321 TI - Impact of NO-synthase inhibition on renal hemodynamics in normotensive and hypertensive subjects. AB - OBJECTIVE: To examine the acute effects of NO-synthase inhibition on renal hemodynamics in normotensive and hypertensive subjects. METHODS: Changes of renal plasma flow (RPF) and glomerular filtration rate (GFR) in response to intravenous infusions of NG-monomethyl-l-arginine (l-NMMA) (3 mg/kg per 30 min) were measured in 32 normotensive and in 39 essential hypertensive patients by use of clearance technique. RESULTS: l-NMMA significantly decreased RPF in normotensive and hypertensive individuals (P < 0.001), while GFR was preserved. Changes of renal hemodynamic parameters were similar in hypertensive and normotensive subjects (deltaRPF: -88 +/- 89 versus -81 +/- 105 ml/min, P = NS; deltaGFR 1.6 +/- 8.2 versus 4.3 +/- 8.9 ml/min, P = NS) Furthermore, l-NMMA increased mean arterial pressure (deltaMAP 5.3 +/- 6.3 versus 6.0 +/- 6.1 mmHg, P = NS) and decreased heart rate (deltaHR -5.8 +/- 3.9 versus -4.1 +/- 3.8 beats/min, P = NS) to a similar extent in both groups. CONCLUSION: Basal NO synthesis of the renal vasculature is not impaired in patients with established essential hypertension. PMID- 11875322 TI - Relations of pulse pressure and other components of blood pressure to preclinical echocardiographic abnormalities. AB - OBJECTIVE: To evaluate the extent to which pulse pressure (PP) is associated with echocardiographic abnormalities, and in particular to whether PP is related to LV hypertrophy taking into account other blood pressure (BP) components. DESIGN: Cross-sectional. SETTING: University hospital, hypertension outpatient unit. PARTICIPANTS: A total of 275 adults (mean age 47 years, range 19-69, 3% aged > or = 65) with essential hypertension. Overt coronary artery disease, valvular disease and secondary hypertension were exclusion criteria. Subjects were divided in two groups with PP < or = 50 or PP > 50 mmHg. OUTCOME MEASURES: Left ventricular (LV) mass, hypertrophy, LV systolic dysfunction. RESULTS: Prevalence of LV hypertrophy was higher in subjects with clinic PP > 50 mmHg. Subjects with PP > 50 mmHg had higher clinic and ambulatory systolic than subjects with PP < or = 50 mmHg while diastolic BP did not differ between groups. PP and systolic BP, either clinic or ambulatory, showed similar correlation to LV hypertrophy in separate logistic multivariate models. Using different methodologies, PP was not related to LV mass index or hypertrophy when the effect of its component systolic BP was taken into account. In separate analyses, PP was not significantly related to ejection fraction or midwall mechanics. CONCLUSION: Middle-aged clinically healthy hypertensives with PP > 50 mmHg had two-fold higher prevalence of LV hypertrophy than those with PP < or = 50 mmHg, which may contribute to the higher cardiovascular risk in subjects with higher PP. However, in our sample, PP was not related to LV hypertrophy independently of systolic BP, suggesting that systolic BP is the explanatory link of the relation between PP and LV hypertrophy. PMID- 11875323 TI - Ethnic differences in carotid and left ventricular hypertrophy. AB - OBJECTIVES: Afro-Caribbean subjects have a higher prevalence of hypertension, a lower prevalence of ischaemic heart disease and a higher premature mortality compared to White Europeans. Left ventricular hypertrophy (LVH) is also more prevalent in Afro-Caribbeans even at similar levels of blood pressure. It is widely believed that carotid artery intima-media thickening (IMT) represents an early marker for the development of atheroma, and carotid IMT and LVH are associated in White populations. Whether the relationship between carotid IMT and LVH is similar in Black subjects is unknown. METHODS: Thirty-eight subjects were studied using carotid and femoral ultrasonography and echocardiography; 19 Afro Caribbean and 19 White European subjects were matched for age, sex and mean 24 h systolic blood pressure. RESULTS: The Afro-Caribbean group had a significantly greater left ventricular mass index (LVMI) compared to the White European: 136.4 +/- 6.1 versus 112.4 +/- 6.2 g/m2, P < 0.01. However, carotid IMT, carotid diameter, femoral IMT and femoral diameter were similar between the groups: 0.75 +/- 0.02 versus 0.77 +/- 0.04 mm, 6.54 +/- 0.15 versus 6.56 +/- 0.16 mm, 0.66 +/- 0.03 versus 0.68 +/- 0.03 mm and 8.40 +/- 0.33 versus 8.25 +/- 0.23 mm, respectively. CONCLUSIONS: Afro-Caribbean subjects with similar blood pressures have similar mean carotid and femoral IMTs compared to White Europeans, in spite of marked differences in LVMI. Whether this reflects a discrepancy in the degree of cardiovascular risk for similar levels of LVMI or whether this is a reflection of an altered pattern of target organ damage associated with hypertension in Afro Caribbean subjects is unclear. PMID- 11875324 TI - Twenty-four hour ambulatory blood pressure in the International Nifedipine GITS Study Intervention as a Goal in Hypertension Treatment (INSIGHT). AB - OBJECTIVES: The International Nifedipine GITS Study Intervention as a Goal in Hypertension Treatment (INSIGHT) showed, by means of office blood pressure measurements, that long-term treatment with nifedipine GITS is as effective as diuretics in preventing cardiovascular and cerebrovascular complications. However, since office blood pressure measurements reflect to a limited extent blood pressure outside the office, a side-arm INSIGHT study in which patients underwent both office measurement and 24 h ambulatory blood pressure monitoring was also performed. DESIGN AND METHODS: The study had a randomized, double-blind, parallel group design. After 4 weeks of placebo, mild-to-moderate essential hypertensive patients were randomized to nifedipine GITS 30 mg or amiloride 2.5 + hydrochlorothiazide 5 mg for 3.1 years. Dose titration was performed by dose doubling and addition of atenolol 25-50 mg or enalapril 5-10 mg, or other drugs when needed. Analysis was carried out by intention-to-treat and included computation of 24 h, day and night ambulatory blood pressure and heart rate values. Additional analyses included computation of the trough-to-peak ratio and the smoothness index (the ratio between the average of the 24-hourly blood pressure reductions after treatment and its standard deviation). RESULTS: A total of 151 patients were recruited and 149 were valid for analysis: 78 patients had 24 h ambulatory recordings both at baseline and during treatment and 134 during treatment. Office, 24 h and day and night blood pressures were all significantly and similarly reduced by both treatments. Office and ambulatory heart rate was left unchanged by diuretics, while it was slightly reduced by nifedipine. Median trough-to-peak ratios were always > 0.5 and superimposable between the two treatment groups. Similarly, smoothness indices of systolic and diastolic blood pressures were comparably high for nifedipine and diuretics, thus demonstrating a similar well-balanced antihypertensive response to both drugs. No significant differences were observed between the two treatment groups in the number of cardiovascular events (17 in the nifedipine-based and 26 in the diuretics-based treatment group). CONCLUSIONS: In the INSIGHT study, the long-term antihypertensive effect on 24 h blood pressure and the cardiovascular protection of nifedipine was similar to that of diuretics. PMID- 11875325 TI - Seeing the forest through the trees: big trucks-little trucks. PMID- 11875326 TI - Hormone replacement therapy and breast tissue density on mammography. PMID- 11875327 TI - Management of postmenopausal osteoporosis: position statement of the North American Menopause Society. AB - OBJECTIVE: The North American Menopause Society (NAMS) established a goal to create an evidence-based position statement regarding the management of postmenopausal osteoporosis. DESIGN: NAMS followed the general principles established for evidence-based guidelines to create this document. A MEDLINE search was conducted. Clinicians and researchers acknowledged to be experts in the field of osteoporosis were enlisted to review the evidence. The NAMS Board of Trustees reviewed and approved the final document. RESULTS: Osteoporosis, which has its highest rate of occurrence in postmenopausal women, increases the risk for fractures, including hip and spine fractures. These injuries are often associated with particularly high morbidity and mortality. Given the health implications of osteoporotic fractures, the primary goal of osteoporosis therapy is to prevent fractures by slowing or preventing bone loss, maintaining bone strength, and minimizing or eliminating factors that may contribute to falls. The evaluation of postmenopausal women for osteoporosis risk requires the recording of a medical history, a physical examination, and diagnostic tests. Major risk factors for osteoporosis are age, genetics, lifestyle (especially nutrition), and menopausal status. Management focuses first on nonpharmacologic measures, such as a balanced diet including adequate calcium and vitamin D intakes, appropriate exercise, smoking cessation, avoidance of excessive alcohol intake, and fall prevention. If pharmacologic therapy is indicated, FDA-approved options are estrogens (prevention only), bisphosphonates and selective estrogen-receptor modulators (prevention and treatment), and calcitonin (treatment only). CONCLUSIONS: Management of postmenopausal osteoporosis involves identifying the potential risk for osteoporosis and osteoporotic fracture, followed by measures that focus on reducing modifiable risk factors through lifestyle changes and, if indicated, pharmacologic therapy. PMID- 11875328 TI - Transdermal estradiol reduces plasma myeloperoxidase levels without affecting the LDL resistance to oxidation or the LDL particle size. AB - OBJECTIVE: This study was designed to investigate the effects of different therapeutic range doses of transdermal estradiol (E(2)), alone or in combination with progesterone (P) or medroxyprogesterone acetate (MPA), on plasma lipoprotein levels and on three parameters related with LDL oxidizability, the resistance of LDL to oxidation by copper, the LDL particle size, and the myeloperoxidase levels. DESIGN: Thirty-five healthy postmenopausal women who had been amenorrheic for at least 1 year received two consecutive, 2-month doses of transdermal estrogen (25-microg and 50-microg E(2) patch). Thereafter, they were randomly assigned to receive a 2-month treatment of either a 100-microg E(2) patch or a 50 microg E(2) patch combined with P (300 mg/day) or MPA (5 mg/day) during the last 14 days. RESULTS: Neither transdermal E(2) alone nor transdermal E(2) plus progestogen modified the lipoprotein profile, the LDL resistance to oxidation, or the LDL particle size. However, all treatments similarly reduced the myeloperoxidase protein levels. CONCLUSIONS: Different dosages of transdermal E(2) within the therapeutic range were equally effective in reducing myeloperoxidase protein levels. The effect remained after addition of P or MPA in a sequential regime. PMID- 11875329 TI - Mammographic changes associated with raloxifene and tibolone therapy in postmenopausal women: a prospective study. AB - OBJECTIVE: The prolonged use of estrogen therapy is associated with a slightly increased risk of breast cancer. Alternative therapies that are effective in the prevention of menopause, having associated morbidities but no unwanted effects, are of primary interest in the pharmacologic research. The aim of this study was to evaluate the effect of two alternative to estrogens drugs, the selective estrogen receptor modulator raloxifene and the tissue-specific tibolone, on the mammographic appearance of the breast. DESIGN: The study group comprised 131 postmenopausal women aged 41 to 67 years. The women were at least 2 years postmenopausal, free of climacteric symptoms, and at the time of entry to the study had not had therapy for at least 9 months. Women with risk factors for osteoporosis or cardiovascular disease were allocated either to tibolone (n = 56) or raloxifene (n = 48) therapy. Women with no risk factors and women who either did not qualify for or denied treatment (n = 27) served as controls. The study duration was 12 months. Women received a baseline mammogram before commencing therapy and a repeat mammogram at the end of the study period. Mammogram findings were classified according to the modified Wolfe criteria by two expert radiologists. RESULTS: No difference was identified between groups with respect to baseline characteristics associated with breast cancer risk. Similarly, no difference was detected between groups concerning the modified Wolfe classification of baseline mammographic findings. In the tibolone group, 10.7% of the women showed an increase in breast density in the 12-month reevaluation. The respective figure in the raloxifene group was 6.3%, whereas no woman in the control group showed an increase in breast density. Differences in the increase in breast density between groups did not, however, reach statistical significance. Accordingly, 10.7% of women in the tibolone group and 18.8% of women in the raloxifene group exhibited involutionary changes in the repeat mammogram, whereas 25.9% of women in the control group revealed a decrease in breast density in the 12-month examination. The percentages were not significantly different between groups. CONCLUSIONS: Breast density as shown by mammography was stable in a majority of patients and changed in a minority of cases for both tibolone and raloxifene. In most patients, these drugs are not likely to interfere with mammogram interpretation. Larger long-term studies are needed to confirm the impact of prolonged tibolone or raloxifene administration on mammography. PMID- 11875330 TI - The effect of hormone replacement therapy on appendicular lean tissue mass in early postmenopausal women. AB - OBJECTIVE: The impact of hormone replacement therapy (HRT) on skeletal muscle mass is still a controversial issue in women's health. Some authors hypothesize anabolic effects, others catabolic. These hypotheses, however, await confirmation by longitudinal observations based on more direct measurements of muscle mass. The aim of the present preliminary study was to evaluate the effect of a 3-year HRT program on appendicular lean tissue mass (LTM(A)) in early postmenopausal women aged 45-54 years. DESIGN: This was a randomized, double-blind and placebo controlled trial. Women received HRT with 2 mg estradiol valerate combined either continuously with 1 mg cyproterone acetate (days 1-28; n = 15) or sequentially with 75 mug levonorgestrel (days 17-28; n = 15), or placebo (n = 18). Serum estradiol was measured by radioimmunoassay. LTM(A) was measured by dual photon absorptiometry (baseline) and dual energy X-ray absorptiometry (years 2 and 3). RESULTS: Baseline serum estradiol did not show significant correlation with the respective LTM(A) (r = 0.018, p = 0.88, n = 75). Cross-sectional analysis found no significant differences between the intervention groups at any time points. The longitudinal changes between years 2 and 3 showed a trend toward decreasing LTM(A) in those receiving HRT (-0.08 +/- 0.12 kg, n = 30) compared to those receiving placebo (0.12 +/- 0.25 kg, n = 18, p = 0.44). CONCLUSIONS: The present preliminary study did not find significant effects on LTM(A) caused by HRT. The trends toward decreasing LTM(A) in the HRT groups might suggest catabolic rather than anabolic effects. These trends, however, await confirmation by larger clinical trials. PMID- 11875331 TI - Sex hormone replacement therapy reverses altered venous contractility in rats after pharmacological ovariectomy. AB - OBJECTIVE: Female sex hormones have several important effects on the venous system. We earlier found that hormone replacement has a significant effect on venous distensibility, but effects of menopause and hormone replacement on venous contractility have never been studied. Therefore, and because the changes we found earlier in distensibility were most likely caused by alterations of contractility, we examined the changes in contractility of saphenous vein caused by depletion and replacement of sex hormones in female rats. DESIGN: Twenty Sprague-Dawley rats were pharmacologically ovariectomized by triptorelin. Ten of these rats received combined sex hormone replacement (HRT) with estradiol propionate and medroxyprogesterone acetate. The rest were given vehicle. Ten animals without ovariectomy served as controls. After 3 months of treatment, segments of the saphenous vein were dissected. Pressure-diameter curves were recorded in relaxed, contracted, and control states. RESULTS: Venous diameter, adjusted for body weight, was significantly decreased after pharmacological ovariectomy. HRT increased the diameter. The presence of sex hormones augmented norepinephrine contraction measured at physiological pressures (control: 19.2 +/- 2.3%; pharmacological ovariectomy: 15.2 +/- 1.4%, p < 0.05 and 17.8 +/- 2.2% following HRT). Myogenic (spontaneous) tone of the saphenous vein did not change after ovariectomy, but it was lowered by hormone replacement (control: 8 +/- 1.1%; ovariectomy: 6.9 +/- 2.5%; ovariectomy + HRT: 2.7 +/- 1.1%, p < 0.05). CONCLUSIONS: Sex hormone depletion induces significant alterations in contractility of the saphenous vein, which could perturb venous capacitance function and distensibility. This effect has a potential role in the development of hypertension and venous varicosity, and these changes could possibly be prevented by HRT. PMID- 11875332 TI - Hormone replacement in postmenopausal women: impact of progestogens on autonomic tone and blood pressure regulation. AB - OBJECTIVE: Depressed heart rate variability (HRV) reflects an imbalance of autonomic tone and independently predicts increased cardiovascular risk in patients with congestive heart failure or after acute myocardial infarction. While hormone replacement therapy (HRT) with estrogens beneficially modulates autonomic tone and blood pressure (BP) regulation in postmenopausal women, the impact of concomitant treatment with progestogens remains unclear. DESIGN: In this cross-sectional study, HRV and BP were examined in 62 healthy women (ages 48 71 years) using digital beat-to-beat interval recordings of heart rate and 24 hour ambulatory BP measurements. RESULTS: Demographic parameters did not differ among women without HRT (n = 23), on estrogen (n = 17; ERT), or on progestogen estrogen containing HRT (n = 22; PERT). Total power of HRV was significantly lower, whereas mean heart rate (HR) was significantly higher among women on PERT group versus controls and ERT (total power: 1611 +/- 146 vs. 2497 +/- 308 and 2472 +/- 348 ms(2); heart rate: 80.7 +/- 1.2 vs. 75.0 +/- 1.4 and 74.0 +/- 2.2 bpm; p < 0.05). In addition, low-frequency power and time-dependent parameters of HRV were lower among women on PERT group versus controls and ERT (p < 0.05). ERT use was associated with reduced systolic and diastolic daytime BP, whereas no significant differences were evident PERT users compared with controls. CONCLUSIONS: Progestogen-containing replacement therapy was associated with increased HR and an attenuation of HRV in postmenopausal women. BP was lower in women on ERT, whereas this effect was offset in the PERT group. These observations could at least partially explain the ambiguous results of progestogen-containing HRT on cardiovascular risk in the Heart and Estrogen/Progestin Replacement Study (HERS). PMID- 11875333 TI - Low dose transdermal estradiol/norethisterone acetate treatment over 2 years does not cause endometrial proliferation in postmenopausal women. AB - OBJECTIVE: We investigated the effects of 2-year transdermal continuous combined estradiol (0.025 mg/day) and norethisterone acetate (0.125 mg/day) (Estragest TTS) on bleeding and on the endometrium. DESIGN: This double-blind, randomized, multicenter, parallel, 1-year trial enrolled 266 healthy women at least 2 years past menopause with intact uteri. Patients received a transdermal patch delivering either 0.025 mg estradiol and 0.125 mg norethisterone acetate daily or placebo. Of the 266 women initially included, 135 (96 Estragest TTS, 39 placebo) completed a second year open follow-up, where all women had the estradiol/norethisterone patch. Endometrial biopsies were performed at weeks 0, 48 (n = 171), and 96 (n =109). Effects on endometrial morphology and uterine bleeding were studied. RESULTS: The overall incidence of endometrial hyperplasia after treatment with the estradiol/norethisterone acetate patch for one year was 0.8% with only one case of atypical hyperplasia. There were no clinically significant changes in endometrial thickness in either treatment group. The proportion of bleed-free patients with the estradiol/norethisterone acetate transdermal system increased from 55% in cycles 1-3 to 83% in cycles 10-12. By the 12th cycle, 92% of patients receiving estradiol/norethisterone acetate patches were bleed-free. No additional hyperplasia was seen during the second year follow-up. CONCLUSIONS: A continuous combined transdermal patch delivering 0.025 mg estradiol/day and 0.125 mg norethisterone acetate/day provided good endometrial protection. The dose maintained a consistently high rate of amenorrhea in postmenopausal women. PMID- 11875334 TI - Estrogenic activity of herbs commonly used as remedies for menopausal symptoms. AB - OBJECTIVE: Women are increasingly turning to herbal therapies in an effort to manage their menopausal symptoms. In this study, we investigate the estrogenic activity of four selected herbs commonly used in menopause, namely dong quai, ginseng, black cohosh, and licorice root. DESIGN: We investigated the effect of these selected herbs on cell proliferation of MCF-7 cells, a human breast cancer cell line. We also assessed their estrogenic activity in a transient gene expression assay system using HeLa cells co-transfected with an estrogen dependent reporter plasmid in the presence of human estrogen receptor ER alpha or ER beta cDNA. Finally, we investigated the estrogenic activity of these herbs using a bioassay in mice. RESULTS: Dong quai and ginseng both significantly induced the growth of MCF-7 cells by 16- and 27-fold, respectively, over that of untreated control cells, while black cohosh and licorice root did not. The herbs tested failed to show transactivation of either hER alpha or hER beta and had no effect on uterine weight in vivo when administered orally to mice for a period of 4 days. CONCLUSIONS: Our studies show that dong quai and ginseng stimulate the growth of MCF-7 cells independent of estrogenic activity. Because of the lack of efficacy and the potential for adverse effects, use of these herbs in humans warrants caution pending further study. PMID- 11875335 TI - Developing a virtual reality environment in petrous bone surgery: a state-of-the art review. PMID- 11875336 TI - Endoscopically assisted minimally invasive microvascular decompression of hemifacial spasm. AB - OBJECTIVE: The aim of this study was to assess the use of endoscopy in minimally invasive surgery of the cerebellopontine angle in cases of hemifacial spasm. MATERIALS AND METHODS: Eighty patients with hemifacial spasm underwent endoscopically assisted microvascular decompression between October 1992 and October 1998, at the Ear, Nose, and Throat Department of Nord Hospital in Marseille, France. The microvascular decompression was performed via a minimally invasive retrosigmoid approach. The cerebellopontine angle was then explored by a 30-degree endoscope to visualize the root exit zone of the facial nerve and the precise location of the site of the conflict. Microvascular decompression was performed under the microscope. If the site was an artery, a Teflon sponge was inserted; if the site was a vein, it was coagulated and then dissected away from the facial nerve. RESULTS: In 80 patients seen regularly for at least 1 year of follow-up, and including patients operated on once or twice, the procedure was successful in 92.5% of patients, brought about improvement in 3.75% (96.25% success plus improvement), and failed in 3.75%. In relation to the type of conflict, success or improvement was experienced by 90.7% of patients with simple conflicts, 86.2% of patients with multiple conflicts, and 87.5% of patients with nutcracker conflicts. No major postoperative complication or mortality occurred in this series. No facial paresis or paralysis occurred immediately postoperatively. Three patients (3.25%) experienced delayed facial palsy. Postoperative cerebrospinal fluid leak occurred in 2 patients (2.5%) and was treated surgically. CONCLUSION: The principle of minimally invasive surgery in the cerebellopontine angle is gaining universal acceptance. The use of endoscopy in microvascular decompression for hemifacial spasm has helped tremendously in improving the results. In this study, the use of the endoscope enabled the authors to identify the site of the conflict in all cases, and to confirm the position of the Teflon sponge before closure. PMID- 11875337 TI - Technique of endoscope-aided myringoplasty. PMID- 11875338 TI - Endoscopy as a tool in minimally invasive trigeminal neuralgia surgery. AB - OBJECTIVE: The aim of this study was assessment of the use of endoscopy in minimally invasive surgery of the cerebellopontine angle in cases of trigeminal neuralgia. METHODS: This study comprises 42 cases of trigeminal neuralgia that underwent operation with endoscopic-assisted microvascular decompression between October 1992 and October 1998. This study was performed in the Ear, Nose, and Throat Department, Nord Hospital, in Marseille, France. The decompression was performed by means of a minimally invasive retrosigmoid approach without a cerebellar retractor. The cerebellopontine angle was then explored by a 30-degree endoscope that gives a panoramic view of this space, with clear visualization of the trigeminal nerve from the pons to Meckel's cave, allowing for the identification of the precise location of the site of the conflict. Microvascular decompression was performed under the microscope by separating the offending vessel from the trigeminal nerve; separation was maintained by the insertion of a piece of Teflon. RESULTS: The site of conflict was detected at the root entry zone of the nerve in 35 patients (83.3%) and at Meckel's cave in 7 patients (16.7%). In 32 cases (76.2%), the type of contact between the vessel and the nerve was of the simple type (1 vessel coming in contact with the nerve in a single point); in 6 cases (14.3%), it was a multiple type (2 vessels touching the nerve in the same point); and in 4 cases (9.5%), it was a nutcracker type (2 vessels compressing the nerve between them). After at least 1-year follow-up and a single operation (cases that required a second operation for revision were considered failures), a successful result was obtained in 31 cases (73.8%), and an improvement was obtained in 4 cases (9.5%). The operation was a failure or early recurrence occurred in 7 cases (16.7%). Postoperative complications were rare. A cerebrospinal fluid leak occurred in only 1 case (2.4%) and was subsequently treated with lumbar puncture and a compressive bandage. CONCLUSION: The minimally invasive retrosigmoid endoscopic-assisted microvascular decompression is an acceptable treatment of primary trigeminal neuralgia. Endoscopy provides a unique way to explore the cerebellopontine angle and to identify the exact location of the neurovascular conflict. PMID- 11875339 TI - Magnetic resonance imaging-based virtual endoscopy of inner ear pathology. AB - OBJECTIVE: To study the feasibility of high-resolution magnetic resonance imaging (MRI)-based virtual endoscopy of the labyrinth to assess subtle inner ear pathology. STUDY DESIGN: A retrospective case review of patient with known inner ear pathology to determine the feasibility and clinical value of MRI-based virtual labyrinthoscopy. SETTING: Tertiary referral center. PATIENTS: Ten patients with symptoms of sensorineural hearing loss or vertigo who underwent high-resolution MRI between 1996 and 1999. INTERVENTION: Diagnostic image modality with three-dimensional (3-D) postprocessing to assess inner ear pathology. MAIN OUTCOME MEASURES: To evaluate how 3-D rendering with virtual labyrinthoscopy can depict subtle labyrinthine pathology. RESULTS: Cases with typical 3-D models and virtual labyrinthoscopic views are presented to illustrate this new image processing approach. CONCLUSION: The virtual endoscopic view of the labyrinth revealed subtle inner ear pathology. This 3-D postprocessing technique is able to render inner surface changes of tiny structures within the inner ear. It can be performed within a very short time using dedicated hybrid rendering techniques. It allows visualization of pathology in a comprehensive way for clinicians and is able to add 3-D information for troubleshooting in doubtful two-dimensional findings. We suggest the term virtual labyrinthoscopy for virtual intraluminal visualization of the labyrinth. PMID- 11875340 TI - Retrosigmoid approach for small and medium-sized acoustic neuromas. AB - OBJECTIVE: Clinical study of the keyhole acoustic neuroma retrosigmoid approach for facial nerve and hearing preservation. STUDY DESIGN: This was a prospective case review from October 1993 to December 1998 in a referral hospital care unit. PATIENTS: A total of 119 consecutive patients with a tumor size of <25 mm in the cerebellopontine angle corrected by a retrosigmoid approach were included in the study. INTERVENTIONS: Standard audiometric and imaging assessments, complete tumor removal by using endoscopy-assisted control, and nerve monitoring. MAIN OUTCOME MEASURES: House-Brackmann facial nerve grade and hearing level by the American Academy of Otolaryngology-Head and Neck Surgery classification. RESULTS: Grades I and II facial nerve function was obtained in 96% of cases, measurable hearing was preserved in 49% of cases, and 30% of cases achieved serviceable hearing. CONCLUSION: The retrosigmoid approach is a safe and reliable approach in random patients with small and medium-sized acoustic neuromas. PMID- 11875341 TI - Effect of topical 5-fluorouracil on closure time of myringotomies created by a radiofrequency surgical unit in guinea pigs. AB - PURPOSE: To delay the closure time of myringotomy without inserting a ventilation tube by creating a myringotomies in guinea pigs by a radiofrequency surgical unit and using topical 5-fluorouracil (5-FU). BACKGROUND: Topical 5-FU was used successfully in glaucoma surgery to depress the proliferation of fibroblasts, but the effects of 5-FU in the myringotomy site in delaying closure are not known. The availability of the use of a radiofrequency surgical unit in performing myringotomy and the effects of a radiofrequency surgical unit myringotomy in the patency period of the myringotomy opening are not known. METHODS: A myringotomy in the tympanic membranes of 30 guinea pigs was created by using a radiofrequency surgical unit. Right ears of these guinea pigs were identified as the study group and sponges with 50 mg/ml 5-FU were applied topically. Saline-soaked sponges were applied to the left myringotomy site, which served as the control side for 15 minutes. Guinea pigs were separated into 2 groups. In Group 2, an additional 20 mg/ml 5-FU and saline solution were dropped into the right and left external ear on the fourth and on the 11th day, respectively. RESULTS: In all guinea pigs, it was found that all the left-ear myringotomy sites were closed at the end of the first week. In the right ears, the myringotomy sites were closed in 75% of Group 1 and in 80% of Group 2 at the end of the second week. In the histopathologic examination of tympanic membranes, there was no apparent difference between study and control sides, but a slight increase in inflammatory findings were encountered in the study sides. CONCLUSION: The closure time of the myringotomy site was delayed twice as much as in the control group in the site where topical 5-FU was applied, and the drop form of 5-FU did not create any additional delay in the closure time. PMID- 11875342 TI - Laser stapedotomy minus prosthesis (laser STAMP): absence of refixation. AB - OBJECTIVE: To determine what percentage of patients with otosclerosis could successfully undergo a laser stapedotomy minus prosthesis over a 5-year period, and to determine the percentage of patients in whom refixation develops during follow-up. STUDY DESIGN: Retrospective case review of 136 patients (137 ears) who underwent primary surgery for otosclerosis. SETTING: An otology/neurotology tertiary referral center. PATIENTS: Patients were chosen if they had clinical evidence of otosclerosis without a history of otologic surgery. INTERVENTIONS: A standard stapes approach was used for all patients. For the laser stapedotomy minus prosthesis, a hand-held laser probe was used to vaporize the anterior crus of the stapes and perform a linear stapedotomy across the anterior one third of the footplate. If otosclerosis was confined to the fissula ante fenestram, the stapes became completely mobile. The stapedotomy opening was sealed with an adipose tissue graft from the ear lobe. MAIN OUTCOME MEASURES: Pure-tone audiometry with appropriate masking and auditory discrimination testing was performed before surgery, 6 weeks after surgery, and every year thereafter. RESULTS: Of the 137 cases, favorable anatomy and minimal otosclerosis allowed 46 (33.6%) of these patients to undergo laser stapedotomy minus prosthesis. Fifty seven patients (41.6%) could not undergo the procedure because of extensive otosclerosis. The remaining 34 patients (24.8%) did not receive laser stapedotomy minus prosthesis because of other anatomic or technical difficulties. Of the 34 patients in the laser stapedotomy minus prosthesis group with more than 4 months follow-up, the average air-bone gap was closed from a mean of 22 dB (SD 10 dB) to 6 dB (SD 4 dB) 6 weeks postoperatively. Follow-up periods ranged from 5 months to 53 months (mean 767 days, SD 437 days). The long-term air-bone gap improved slightly to an average of 5 dB (SD 6 dB) in comparison with the sixth postoperative week value. CONCLUSION: Laser stapedotomy minus prosthesis is a minimally invasive procedure, which over the follow-up period has a very low incidence of refixation, as evidenced by a lack of progressive conductive hearing loss. The success of this procedure depends on the correct selection of cases. This procedure has been successfully performed on 33.6% of patients undergoing primary stapes surgery. Laser stapedotomy minus prosthesis seems to be a viable alternative to conventional stapedotomy that yields good results without evidence of refixation over an extended time. PMID- 11875343 TI - Otologic manifestations of amyloidosis. AB - OBJECTIVE: To describe otologic manifestations of amyloidosis. STUDY DESIGN: Retrospective case review. SETTING: Tertiary referral center. PATIENTS: Case report of a patient with bilateral external auditory meatal stenosis secondary to amyloidosis. RESULTS: The patient is a 60-year-old man who presented with several months' history of bilateral ear blockage and drainage. Physical examination revealed markedly thickened skin in each ear in the area of concha and meatus that was associated with fissuring. The meatus were markedly stenosed. The medial aspect of the external auditory canal and the tympanic membrane were normal. He had partial improvement on medical therapy with topical steroid-containing eardrops and ointment. Past medical history was positive for multiple myeloma. Meatal biopsy was performed. Grossly, the tissue was hypovascular and markedly thickened. Histopathologic examination was consistent with amyloidosis. The patient was continued on topical steroid ointments, frequent ear cleaning, and close follow-up. He was subsequently diagnosed with systemic amyloidosis. A review of the literature was done. Head and neck involvement with amyloidosis is reviewed, with emphasis on otologic manifestations. CONCLUSION: Otologic involvement with amyloidosis is extremely rare. The pathophysiology may be related to the local factors in the external auditory canal. PMID- 11875344 TI - Infection screening in sudden and progressive idiopathic sensorineural hearing loss: a retrospective study of 182 cases. AB - OBJECTIVE: To evaluate a systematic infectious screening program for patients who present with sudden or progressive sensorineural hearing loss (SHL) of unknown cause (negative history and clinical examination). METHOD: Retrospective study of 182 patients with idiopathic SHL. One hundred six patients presented with sudden SHL, and 76 presented with progressive SHL. Serologies for herpes simplex and varicella-zoster viruses (immunoglobulin M and immunoglobulin G titers), Lyme disease (enzyme-linked immunosorbent assay), syphilis (fluorescent treponemal antibody absorption or microhemagglutination-Treponema pallidum test) and human immunodeficiency virus were performed. RESULTS: The serologies were negative in 179 patients. Two patients had positive serologies for Lyme disease and another tested positive for syphilis. Both cases of suspected Lyme disease were later excluded by Western blot analysis and lumbar puncture (two false-positives). The patient with serologic syphilis was diagnosed as having latent syphilis after neurosyphilis was excluded. CONCLUSION: The infection screening was positive in only 1 (0.6%) of 182 patients: the patient who was diagnosed with latent syphilis. On the basis of these results and taking into account the cost of systematic screening, we propose that serologic tests be limited to patients with suspect histories or symptomatologies, except for patients with a diagnosis of syphilis. PMID- 11875345 TI - Pediatric cochlear implantation in auditory neuropathy. AB - OBJECTIVE: Auditory neuropathy (AN) is characterized by varying degrees of sensorineural hearing loss, an absent or severely abnormal auditory brainstem response, and normal otoacoustic emissions. The nomenclature for this condition reflects the concept that the site of lesion is proximal to the cochlea (e.g., cochlear nerve). Given this hypothesis, it is reasonable to expect limited benefit from cochlear implantation in patients with AN. However, a growing body of evidence shows the striking benefits of cochlear implantation in AN. To explore this topic, we reviewed our population of children with AN and, specifically, the performance results in those children having undergone cochlear implantation. STUDY DESIGN: A retrospective case review of those patients diagnosed with AN from 1993 to 2001. SETTING: A tertiary pediatric referral center. PATIENTS: A diagnosis of AN reported from the Center for Hearing and Deafness Research, Cincinnati, OH, database. RESULTS: Eighteen patients were diagnosed with AN (11 girls, 7 boys), with 3 sets of siblings, including 1 set of identical twins. Four patients with AN underwent implantation in the previous 5 years. Twelve out of the 18 patients had classic risk factors for AN (e.g., prematurity and hyperbilirubinemia). The degree of hearing loss varied in our patients, with a majority showing severe to profound deficits. All children with implants showed improvement in auditory and verbal development, but this improvement was variable. CONCLUSION: The success of cochlear implantation in these patients suggests that some children with AN have an auditory system lesion that can be compensated for by cochlear implantation. This implies either an inner hair cell or inner hair cell-cochlear nerve junctional pathology that can be overcome by direct electrical stimulation. PMID- 11875346 TI - Binaural cochlear implants placed during the same operation. AB - OBJECTIVE: To evaluate the binaural listening advantages for speech in quiet and in noise and to localize sound when independently programmed binaural cochlear implants are used, and to determine whether ears with different hearing ability and duration of profound deafness perform differently with cochlear implants as well as to what extent preimplant psychophysical and physiologic assessment could be predictive of performance. STUDY DESIGN: Prospective study in which patients were prospectively selected to undergo bilateral implantation during a single surgical procedure at a tertiary referral center. All testing was performed with patients using their right, left, or both cochlear implants. Preimplant and intraoperative measures used electrical stimulation at the round window and stimulation through the cochlear implant. RESULTS: Bilateral implantation during the same operation did not cause any postoperative problems such as severe vertigo or ataxia. At 1 year, results of speech testing in quiet demonstrated a binaural advantage for 2 of 10 subjects. Speech-in-noise testing demonstrated that two implants were beneficial for two individuals. All subjects benefited from a head shadow effect when an ear with a better signal-to-noise ratio was available. The ability to localize sound was improved with binaural implants in all subjects. Preimplant psychophysical or physiologic measures were not predictive of eventual speech perception performance. CONCLUSION: Binaural cochlear implants can assist in the localization of sounds and have the potential in some individuals to improve speech understanding in quiet and in noise. PMID- 11875347 TI - Temporal bone histopathology related to cochlear implantation in congenital malformation of the bony cochlea. AB - HYPOTHESIS: Histopathologic findings in temporal bones with congenital malformations of the bony cochlea may provide insight into cochlear implantation planning, surgical approach, and complications. BACKGROUND: Patients with congenitally malformed cochleae account for an increasing percentage of candidates for cochlear implantation. Few studies on the relationship between histopathologic findings of temporal bones with malformation of the bony cochlea and cochlear implantation have been reported. METHODS: We studied 21 temporal bones from 12 cases with congenital malformations of the bony cochlea. Ages ranged from stillborn to 50 years. Length of the cochleae and dimensions of facial recesses were measured with light microscopy. Other malformations associated with a shortened cochlea were determined, emphasizing how they affect cochlear implantation. RESULTS: The average length of the malformed cochlear duct was 22.84 +/- 0.69 mm. Average dimensions of facial recesses were not significantly different between malformed cochleae and the controls. Other malformations included enlarged cochlear aqueduct (43%), abnormal facial nerve course (57%), enlarged vestibular aqueduct (52%), aplasia of the middle ear (19%), malformed ossicles (67%), abnormal oval window (57%), and abnormal round window (29%). CONCLUSION: A shortened cochlear duct may cause an incomplete insertion of the implant electrode. Because dimensions of the facial recesses are similar to normal cases, a facial recess surgical approach is recommended. An enlarged cochlear aqueduct may cause perilymphatic oozing or gushing on fenestration of the cochlea. An anomalous course of the facial nerve is a common finding. Tympanic malformations such as round and oval window deformities and small middle ear cleft should be noted preoperatively to better guide surgery. PMID- 11875348 TI - Movements of cochlear implant electrodes inside the cochlea during insertion: an x-ray microscopy study. AB - BACKGROUND: There are no satisfactory, noninvasive techniques currently available to visualize the cochlear implant (CI) electrode in a dynamic state as it is advanced inside the cochlea. OBJECTIVE: This study describes a radiologic technique that can be used in temporal bones to monitor the electrode position in real time and to visualize the basilar membrane. METHODS: A cochleostomy was performed in accordance with the normal procedure for cochlear implantation in seven fresh cadaveric temporal bones. A special x-ray tube (Microfocus; Focus, Wunstorf, Germany) with a spot size of 1 microm and fluoroscopy were used for continuous videotape imaging of the advancing electrode in the cochlea. Conventional electrodes (MED-EL 40+; MED-EL, Innsbruck, Austria) and prototypes of perimodiolar electrodes (MED-EL), with varying thicknesses and angles of insertion, were used for the study. Finally, contrast liquid (Ultravist; Berlex Imaging, Schering, Germany) was injected into the scala vestibuli through the stapes footplate. RESULTS: The advancing electrode carrier was clearly visible at x20 magnification. With the perimodiolar electrodes, deep insertion was impossible if the guiding wire was too rigid, and retraction occurred if the wire was soft. The intact nature of the basilar membrane could be confirmed by the sequential filling of scala vestibuli and the scala tympani. CONCLUSION: To our knowledge, this is the first noninvasive study demonstrating continuous visualization with high-resolution and x-ray magnification of CI electrode advancement inside the intact cochlea. Our technique can be used for the development of CI electrodes with optimal design characteristics, especially stiffness. Visualization of the basilar membrane will influence the insertion techniques when CIs are developed for patients with residual hearing. PMID- 11875349 TI - Fate of the mesenchyme in the process of pneumatization. AB - HYPOTHESIS: This study's aim was to find histologic data that would indicate the mode of disappearance of the embryonal mesenchyme. BACKGROUND: The basic studies made during the first half of the 20th century concluded that mesenchyme disappears by regression and resorption. Recently, it was suggested that mesenchyme disappears by receding, spreading, and thinning to match the enlarging bony spaces. METHODS: We studied 11 serially sectioned temporal bones from newborns to adults and describe detailed findings in a 9-day-old newborn and in a 1.5-year-old infant. The temporal bones were sectioned to 20 mum and stained by hematoxylin and eosin. RESULTS: Histologic evidence of regression was found in the form of degenerating mesenchymal cells and fibers, in areas free of cells, and with empty spaces of varying size between the fibers. Vacuoles differing much in size appeared, and phagocytic cells were frequent. A rich capillary network allowed resorption of hemopoietic cells dispersed from the marrow spaces into the mesenchyme. From the lower lateral attic, from Prussak's space, and from the mastoid air cells, mesenchyme can disappear only by regression-there is no space where it could recede. CONCLUSION: Pneumatization of the middle ear spaces occurs by regression and resorption with an individual speed under genetic guidance. The osteoclastic activity of the periosteum, intertwined with the nearest mesenchyme, is decisive in the mastoid air cell formation. Dispersion and reabsorption of hemopoietic cells is a normal phenomenon in this process. Underpressure in the middle ear spaces, caused either by a meconium-related foreign body otitis media in infancy or by chronic otitis media in childhood, are factors that may lead to a partial or full arrest of pneumatization. PMID- 11875350 TI - Temporal bone anomalies in the branchio-oto-renal syndrome: detailed computed tomographic and magnetic resonance imaging findings. AB - OBJECTIVE: To inventory computed tomographic and magnetic resonance imaging findings in the branchio-oto-renal (BOR) syndrome. STUDY DESIGN: A prospective computed tomographic and magnetic resonance imaging study on a family with the BOR syndrome. SETTING: Department of medical imaging and magnetic resonance imaging at St. Jan Brugge, Brugge, Belgium. PATIENTS: Eight affected members of a Belgian family. Younger affected family members were excluded because of their age. RESULTS: Computed tomography showed inner ear malformations in all eight affected patients. Magnetic resonance imaging was performed on five patients and showed inner ear malformations. To define hypoplasia or congenital enlargement of the inner ear structures, measurements obtained from a control group of normal subjects were used for comparison. Almost symmetrical cochlear abnormalities were observed on the three-dimensional Fourier transformation-constructive interference in steady state images of the five patients who underwent magnetic resonance imaging; four had dysplasia of the cochlea, and one had hypoplasia. The vestibule was slightly enlarged in one patient; computed tomography and magnetic resonance imaging showed semicircular canal malformations. Magnetic resonance imaging clearly showed bilateral enlarged endolymphatic sacs and ducts, whereas computed tomography showed only unilateral widening of the vestibular aqueduct and borderline widening of the vestibular aqueduct. Magnetic resonance imaging showed bilateral hypoplasia of the cochlear branch of the eighth nerve in one patient. CONCLUSION: Hypoplasia and dysplasia of the cochlea were consistent findings, and only magnetic resonance imaging was able to evaluate the intracochlear changes in detail and corrected computed tomography in most patients. Moreover, magnetic resonance imaging also detected bilateral hypoplasia of the cochlear branch of the eighth nerve in one patient. A widened vestibular aqueduct and a widened vestibular sac were frequent but not obligatory features of the BOR syndrome. Other malformations of the middle ear included a reduced middle ear cavity and malformations of the ossicular chain. PMID- 11875351 TI - Three-dimensional Fourier transformation constructive interference in steady state magnetic resonance imaging of the inner ear in patients with unilateral and bilateral Meniere's disease. AB - OBJECTIVE: In this study, three-dimensional Fourier transformation constructive interference in steady state (3DFT-CISS) magnetic resonance imaging was used to quantify the distance between the vertical part of the posterior semicircular canal and the posterior fossa as a measure of the endolymphatic sac and duct in patients with Meniere's disease. Differences in this distance between affected and unaffected ears, as well as differences between unilaterally and bilaterally affected patients, were studied and compared with a control group. Also, possible correlations between the measured distance and the duration and severity of symptoms, patient age, and average hearing loss were investigated in the group of patients with Meniere's disease. STUDY DESIGN: Retrospective clinical study. SETTING: Tertiary referral center (University Hospital) as part of a large, diagnostic research project on Meniere's disease. PATIENTS: Of the 111 patients with Meniere's disease initially included, 90 patients underwent 3DFT-CISS MRI. Eighty-six of these patients were analyzed in this MRI study. Fifty-six patients had unilateral Meniere's disease, and 30 patients had bilateral Meniere's disease (116 affected and 56 unaffected ears). Sixty-two ears in patients without Meniere's disease were studied as controls. INTERVENTION: The distance between the vertical part of the posterior semicircular canal and the posterior fossa was determined by 3DFT-CISS MRI. MAIN OUTCOME MEASURES: Contiguous axial 3DFT-CISS MRI slices of 0.7 to 1.0 mm were made by a radiologist according to a strict protocol. Measurements of the distance between the vertical part of the posterior semicircular canal and the posterior fossa were taken by two professionals-a radiologist and an otolaryngologist-using a ruler and the original scan. RESULTS: A significantly smaller distance (2.9 mm) between the vertical part of the posterior semicircular canal and the posterior fossa as visualized on MRI scans was found in the ears of patients with Meniere's disease than in the ears of patients in the control group (3.8 mm, p < 0.001). In both uni- and bilaterally affected patients (n = 56 and n = 30, respectively), no significant difference between ears was found (p = 0.44 and p = 0.19, respectively). In bilaterally affected patients, however, this distance (3.2 mm) was significantly greater than the distance in unilaterally affected patients (2.7 mm, p = 0.004). There was no relationship between the MRI-visualized distance between the vertical part of the posterior semicircular canal and the posterior fossa and the duration of disease, average hearing loss, or severity of symptoms in uni- and bilaterally affected patients. CONCLUSION: The difference in MRI-visualized distances between the vertical part of the posterior semicircular canal and the posterior fossa of uni- and bilaterally affected patients strongly suggests that unilateral and bilateral hearing loss are two different entities in patients with Meniere's disease. The size of the endolymphatic sac seems not to be the only factor in the pathogenesis of Meniere's disease. That the MRI-visualized distance between the vertical part of the posterior semicircular canal and the posterior fossa does not have any relationship to the duration of the disease or to patient age indicates that this distance is a congenital feature. PMID- 11875352 TI - Can posturography identify informed malingerers? AB - OBJECTIVE: This study evaluated whether information about computed dynamic posturography given to subjects instructed to feign vestibular impairment (malingerers) improved their attempts to display test patterns consistent with vestibular weakness. STUDY DESIGN: The study design was prospective, with blinded scoring of tests. SETTING: The study was conducted at a university hospital. PATIENTS: Sixty volunteer subjects, 20 to 59 years old, were randomly assigned to one of two equal groups: naive subjects and subjects informed about computed dynamic posturography. INTERVENTIONS: Computed dynamic posturography was performed twice. First, subjects were instructed to feign a vestibular weakness. Second, subjects were instructed to perform the test normally. One group was given additional information about computed dynamic posturography before the first test (informed faking). MAIN OUTCOME MEASURES: The computed dynamic posturography test summaries were scored as normal, aphysiologic, or vestibular by two methods: the scoring method published by Cevette et al. in 1995 and blinded clinical scoring. The computed dynamic posturography test summaries of patients referred for vestibular disorders were included with the test summaries collected from subjects during scoring. These additional tests of possible "true" vestibular disorder allowed evaluation of scoring accuracy. RESULTS: The scoring method of Cevette et al. classified about one-third of the test results of feigned vestibular weakness as either normal (16.7%) or vestibular weakness (18.3%). The remaining two thirds of the test results of feigned vestibular weakness were classified as aphysiologic (65.0%). Blinded clinical scoring gave similar figures. Additional information about computed dynamic posturography did not increase the number of feigned test results scored as a vestibular weakness. CONCLUSION: When additional information about computed dynamic posturography was provided, the subjects did not improve their ability to feign vestibular weakness. This investigation supports the use of scoring computed dynamic posturography results by the formulae of Cevette et al. in separating aphysiologic behavior from vestibular impairment. PMID- 11875353 TI - Selective retrosigmoid vestibular neurectomy without internal auditory canal drill-out: an anatomic study. AB - OBJECTIVE: It is well established that selective vestibular nerve section by means of the retrosigmoid or posterior fossa approach can be accomplished with or without drill-out of the internal auditory canal (IAC) by virtue of the presence or absence of a surgically accessible cleavage plane between the vestibular and cochlear nerves. Some reports have indicated that a majority of patients would be amenable to successful separation of the vestibular nerve from the cochlear nerve medial to the IAC, thus obviating the need for IAC drill-out and associated complications. However, other reports have indicated routine difficulty in finding a satisfactory vestibulocochlear cleavage plane within the cerebellopontine angle. This in situ cadaver study was undertaken to determine whether normal anatomic relationships support the hypothesis that selective vestibular nerve section can be accomplished by means of the posterior fossa approach without the need for concomitant IAC drill-out in a majority of circumstances. METHODS: A retrosigmoid approach to the posterior fossa was performed bilaterally on 36 intact human cadavers. After displacement of the cerebellum, an operating surgical microscope was used to visualize the cerebellopontine angle in the surgical position. The ability to develop a satisfactory cleavage plane between the vestibular and cochlear nerves without the need for drill-out of the IAC was established in each case. RESULTS: Seventy two vestibulocochlear nerve bundles in 36 intact human cadavers were analyzed. A vestibulocochlear nerve cleavage plane within the cerebellopontine angle amenable to neurectomy medial to the porus of the IAC was observed in 81% left and 69% right vestibulocochlear nerve bundles (average, 75%). The facial nerve was found deep or anterior to the vestibulocochlear nerve bilaterally in all cases examined. The anterior inferior cerebellar artery, or a branch of the artery, was found to cross the plane between the facial and vestibulocochlear nerve bundles within the lateral cerebellopontine angle in 47% of the cases on the left and in 50% of cases on the right. CONCLUSIONS: A vestibulocochlear nerve cleavage plane amenable for selective vestibular nerve transection without drilling the IAC was found in 75% of the 72 cerebellopontine angles studied. The facial nerve consistently lies deep or anterior to the vestibulocochlear nerve within the cerebellopontine angle with the retrosigmoid approach. These findings support the rational and feasibility of avoiding drill-out of the IAC in the majority of circumstances when performing selective vestibular neurectomy by means of the posterior fossa approach for Meniere's syndrome and other vestibular disorders. PMID- 11875354 TI - Translabyrinthine approach for the management of large and giant vestibular schwannomas. AB - BACKGROUND: The removal of large vestibular schwannomas through the translabyrinthine approach is still controversial. MATERIALS AND METHODS: A retrospective review was performed of 81 patients (58 men and 23 women) with vestibular schwannomas 3 cm or greater, who underwent tumor removal via the translabyrinthine approach between 1985 and 2000. The mean tumor size was 3.7 +/- 0.81 cm, and the mean age of the patients was 47 +/- 16.1 years. The largest tumor was 6 cm. All surgical procedures were performed in collaboration with a neurosurgery team. RESULTS: Total tumor removal was accomplished in 77 cases (95.1%). The facial nerve was preserved anatomically in 69 (85.2%) of the patients. In 4 patients, divided nerves were repaired by primary anastomosis. Facial nerve function was assessed immediately after surgery and 1 year or more after discharge. Good function (House-Brackmann facial nerve Grade I or II) was present in 45% of patients and acceptable function (Grades I-IV) in 80% of patients 1 year after resection of the tumor. Cerebrospinal fluid leakage occurred in 12 patients (17%), meningitis developed in 3 patients (4%), and 1 patient experienced a stroke immediately after surgery. There were no deaths caused by surgery in this series. CONCLUSION: The translabyrinthine approach offers an excellent anatomical view of the cerebellopontine angle and a direct approach to the tumor with functional preservation of the facial nerve. Total removal is accomplished in most cases, with minimum incidence of morbidity and no incidence of mortality. PMID- 11875355 TI - Sigmund Freud and the VIIIth Cranial Nerve. PMID- 11875356 TI - Bilateral semicircular canal aplasia: a characteristic of the CHARGE association. PMID- 11875357 TI - Tuberculous otitis media. PMID- 11875359 TI - Myths in neurotology, revisited: smoke and mirrors in tinnitus therapy. PMID- 11875360 TI - A liberatory maneuver for the treatment of horizontal canal paroxysmal positional vertigo. PMID- 11875361 TI - Linking single nucleotide polymorphisms. PMID- 11875362 TI - CYP2C9 polymorphism: impact on tolbutamide pharmacokinetics and response. PMID- 11875363 TI - Molecular haplotyping of genomic DNA for multiple single-nucleotide polymorphisms located kilobases apart using long-range polymerase chain reaction and intramolecular ligation. AB - Genetic polymorphisms are well-recognized causes of interindividual differences in disease risk and treatment response in humans. For genes containing multiple single-nucleotide polymorphisms (SNPs), haplotype structure is often the principal determinant of phenotypic consequences, and haplotype distribution represents the best approach for assessing patterns of linkage disequilibrium. To permit more widespread molecular determination of haplotypes, we developed a simple yet robust method to determine haplotype structure for multiple SNPs located up to 30 kb apart in genomic DNA using long-range polymerase chain reaction (LR-PCR) and intramolecular ligation. Complete concordance was shown between the new method and conventional approaches, such as family pedigree analysis or cloning and sequencing. The availability of a simple method to directly determine haplotype structure using genomic DNA, without family pedigree analysis, cloning or complex instrumentation, provides an important new tool for elucidating the genetic determinants of drug disposition and effects, disease risk, and molecular evolution. PMID- 11875364 TI - Impact of CYP2C9 and CYP2C19 polymorphisms on tolbutamide kinetics and the insulin and glucose response in healthy volunteers. AB - Tolbutamide is known to be metabolized by cytochrome P450 2C9 (CYP2C9), and the effects of the CYP2C9 amino acid polymorphisms *2 (Arg144Cys) and *3 (Ile359Leu) could be important for drug treatment with tolbutamide and for use of tolbutamide as a CYP2C9 test drug. Tolbutamide pharmacokinetics and plasma insulin and glucose concentrations were studied in 23 healthy volunteers with all six combinations of the CYP2C9 alleles *1, *2 and *3, including two subjects with the combined CYP2C9*1/*1 and CYP2C19*2/*2 genotype. Volunteers received a single oral dose of 500 mg tolbutamide, followed by 75 g oral glucose at 1, 4.5 and 8 h after tolbutamide administration. Pharmacokinetic analysis was performed using a computer program for regression analysis of nonlinear mixed effects models. The mean oral clearances of tolbutamide were 0.97 (95% confidence interval [CI] 0.89 1.05), 0.86 (95% CI 0.79-0.93), 0.75 (95% CI 0.69-0.81), 0.56 (95% CI 0.51-0.61), 0.45 (95% CI 0.41-0.49) and 0.15 (95% CI 0.14-0.16) l/h in carriers of CYP2C9 genotypes 1/*1, *1/*2, *2/*2, *1/*3, *2/*3 and *3/*3, respectively. Tolbutamide pharmacokinetics in carriers of the functionally deficient CYP2C19*2/*2 genotype were not different from those in the CYP2C19 highly active genotype. Elimination in the six CYP2C9 genotype groups could be expressed as the linear combination of three constants (0.05, 0.04, 0.01 h(-1), which were specific to the respective CYP2C9 alleles *1, *2 and *3, thus indicating a co-dominant mode of inheritance. Insulin and glucose concentration-time curves did not change with differing CYP2C9 genotypes. Tolbutamide was confirmed as a substrate of the genetically polymorphic enzyme CYP2C9. The pronounced differences in pharmacokinetics due to the amino acid variants did not significantly affect plasma insulin and glucose concentrations in healthy volunteers. PMID- 11875365 TI - Effects of CYP2C19 and CYP2C9 genetic polymorphisms on the disposition of and blood glucose lowering response to tolbutamide in humans. AB - Several recent in-vitro data have revealed that CYP2C19, in addition to CYP2C9, is also involved in the 4-methylhydroxylation of tolbutamide. We evaluated the relative contribution of CYP2C9 and CYP2C19 genetic polymorphisms on the disposition of blood glucose lowering response to tolbutamide in normal healthy Korean subjects in order to reappraise tolbutamide as a selective in-vivo probe substrate of CYP2C9 activity. A single oral dose of tolbutamide (500 mg) or placebo was administered to 18 subjects in a single-blind, randomized, crossover study with a 2-week washout period. Twelve subjects (of whom six were CYP2C19 extensive metabolizer (EM) and six were CYP2C19 poor metabolizer (PM) genotype) were of the homozygous wild-type CYP2C9*1 genotype; the other six subjects were of the CYP2C9*1/*3 and CYP2C19 EM genotype. Pharmacokinetic parameters were estimated from plasma and urine concentrations of tolbutamide and 4 hydroxytolbutamide. Serum glucose concentrations were measured before and after oral intake of 100 g dextrose. In subjects heterozygous for the CYP2C9*3 allele, C(max) and AUC of tolbutamide were significantly greater and the plasma half-life significantly longer than those in homozygous CYP2C9*1 subjects. No pharmacokinetic differences were found between CYP2C19 EM and PM genotype subjects. The estimated AUC of the increase in serum glucose after oral intake of 100 g dextrose was 2.7-fold higher in subjects with the wild-type CYP2C9 genotype than in those with CYP2C9*1/*3, but CYP2C19 genetic polymorphism did not alter the blood glucose lowering effect of tolbutamide. The plasma AUC of 4 hydroxytolbutamide and the ratio of 4-hydroxytolbutamide/tolbutamide did not differ significantly between CYP2C19 PM and EM genotype subjects, while these parameters were about twice as high in subjects with the wild-type CYP2C9 genotype than in heterozygous CYP2C9*3 subjects (P < 0.05). Our results strongly suggest that the disposition and hypoglycemic effect of tolbutamide are affected mainly by CYP2C9 genetic polymorphism, but not by CYP2C19 polymorphism. The in vivo contribution of CYP2C19 to tolbutamide 4-methylhydroxylation appears to be minor in humans. This suggests that, at least in vivo, tolbutamide remains a selective probe for measuring CYP2C9 activity in humans. PMID- 11875366 TI - Common allelic variants of cytochrome P4503A4 and their prevalence in different populations. AB - Marked interindividual variability in expression of CYP3A4 influences the disposition of many endo- and xenobiotics, including the metabolism of steroids, environmental toxins and therapeutically useful drugs. The present study was designed to determine the genetic basis of CYP3A4 variability. We analysed DNA from 82 individuals with known CYP3A4 phenotype including 53 Caucasians and 21 African-American liver donors, seven individuals who were outliers in CYP3A4 metabolism and five individuals in a family of a poor nifedipine metabolizer. In addition, we analysed DNA from the eight person DNA Polymorphism Discovery Resource subset (Coriell Institute) and 89 individuals representing nine ethnic groups. Five non-synonymous mutations in the coding region of CYP3A4 were observed. CYP3A4*14 (T44C) in exon 1 resulted in an L15P change; CYP3A4*15 (G14387A) in exon 6 resulted in a R162Q substitution; CYP3A4*10 (G14422C) in exon 6 resulted in a D174H substitution; CYP3A4*16 (C15721G) in exon 7 resulted in a T185S amino acid substitution; and CYP3A4*12 (C22002T) in exon 11 resulted in a L373F change in the CYP3A4 protein. An additional six single nucleotide polymorphisms (SNPs) in the 5'-UTR, 13 SNPs in the introns and three SNPs in the 3'-UTR were observed. Extensive population differences were observed in the frequencies of various CYP3A4 alleles. None of the 28 CYP3A4 SNPs identified in CYP3A4 phenotyped persons (most individuals being heterozygous for any CYP3A4 variant) was associated with low hepatic CYP3A4 protein expression or low CYP3A4 activity in vivo. PMID- 11875367 TI - A comparative analysis of translated dihydropyrimidine dehydrogenase cDNA; conservation of functional domains and relevance to genetic polymorphisms. AB - A pharmacogenetic syndrome caused by molecular defects in the dihydropyrimidine dehydrogenase gene (DPYD ) results in partial to complete loss of dihydropyrimidine dehydrogenase (DPD) enzyme activity with patients exhibiting life-threatening toxicity following administration of routine doses of 5 fluorouracil. To date, more than 19 reported mutations have been putatively associated with DPD deficiency with 16 occurring within the open reading frame of the cDNA. The purpose of this study was to examine the conservation of functional domains (including the uracil, flavine adenine dinucleotide and NADPH binding sites) across three phyla (Chordata, Arthropoda and Nematoda) and the conservation of regions corresponding to the previously reported mutations. Comparative analysis of the uracil and NADPH binding sites in mammals and invertebrates demonstrated 100% amino acid identity between mammals and Drosophila melanogaster. Caenorhabditis elegans demonstrated 89% and 88% identity in these domains, respectively. The mammalian sequences demonstrated 100% identity in two iron sulphur motifs (amino acids 953-964 and 986-997) with significant conservation in D. melanogaster (92% and 92% identity, respectively) and C. elegans (100% and 92% identity, respectively). Comparative amino acid analysis revealed non-conservation in the loci of four DPYD mutations [DPYD*12 (R21Q), DPYD*5 (I543V), DPYD*6 (V732I), DPYD*9A (C29R)]. Seven mutations occurred in highly conserved regions [M166V, DPYD*8 (R235W), DPYD*11 (V335l), DPYD*4 (S534N), DPYD*9B (R886H), D949V, DPYD*10 (V995F)]. In summary, this comparative analysis identified conserved regions which may be critical to enzyme structure and/or function. The conservation of loci where DPYD mutations occur was also examined to evaluate their functional significance on DPD enzyme activity. These data should prove useful in the evaluation of newly discovered mutations in the DPYD gene. PMID- 11875368 TI - Genetic polymorphisms in heterocyclic amine metabolism and risk of colorectal adenomas. AB - High red meat intake has been linked with an increased risk of colorectal cancer and adenomas. During high temperature cooking of red meats, heterocyclic amines (HCAs) are generated; however, to be carcinogenic, they must be metabolized by enzymes including cytochrome P450 1A2 (CYP1A2) and N-acetyltransferase 1 (NAT1) and/or N-acetyltransferase 2 (NAT2). We have conducted a clinic-based case control study of colorectal adenomas that focused on assessment of exposure to HCAs (estimated by use of a HCA database and meat cooking module) and modification of these exposures by genetic factors. We have previously reported that intake of MeIQx was associated with an increased risk of colorectal adenomas [overall association at 80th percentile, > 27.00 ng/day: odds ratio (OR) = 2.68, 95% confidence interval (CI) 1.58-4.55]. Here, we report our evaluation of whether variation in CYP1A2, NAT1 and/or NAT2 modify the association between HCAs and colorectal adenoma formation in 146 cases and 228 frequency-matched controls. The NAT1*10 allele was associated with a nonsignificant increased risk of colorectal adenomas (OR = 1.43; 95% CI 0.86-2.36). Further, when we analysed 2 amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx) intake as a categorical variable, we observed a six-fold increase in adenoma risk among rapid NAT1 acetylators who consumed more than 27 ng a day (OR = 6.50; 95% CI 2.16-19.7), whereas among slow NAT1 acetylators, the increase in risk was two-fold (OR = 2.32; 95% CI 1.12-4.81). While suggestive, the results were not significantly different from each other on either an additive or multiplicative scale. In contrast, NAT2 genotype and CYP1A2 and NAT2 hepatic activity measured by caffeine urinary metabolites were not associated with adenoma risk, although an increase in risk with rapid CYP1A2 activity could not be ruled out (OR = 1.46; 95% CI 0.76 2.81). Moreover, there was no evidence that the effect of MeIQx was enhanced among subjects in any subgroup defined by variation in these measures. These results are compatible with the hypothesis that high HCA exposure is associated with an increased risk of colorectal adenomas, particularly in genetically susceptible subgroups. Further study of larger populations is needed to confirm and extend these observations. PMID- 11875369 TI - Sequence variation and phylogenetic history of the mouse Ahr gene. AB - The Ahr locus encodes for the aryl hydrocarbon receptor (AHR), which plays an important toxicological and developmental role. Sequence variation in this gene was studied in 13 different mouse lines that included eight laboratory strains, two Mus musculus subspecies and three additional Mus species. The data presented represent the largest study of sequence variation across multiple mouse lines in a single gene (approximately equal to 15.9 kb/mouse line). Among all mice, the average frequency of all polymorphisms in the intronic regions was 20.3 variants/kb and the average exonic frequency was 14.1 variants/kb. For substitutions alone, the average frequencies in the intronic and exonic regions for all mice were 13.3 and 8.9 substitutions/kb, respectively. Between laboratory strains, the average intronic and exonic frequencies for all polymorphisms dropped to 5.4 and 2.9 variants/kb, respectively. There were 111 non-synonymous polymorphisms that resulted in 42 different amino acid changes, of which only 10 amino acid changes had been previously identified. Based on the nucleotide sequence, the phylogenetic history of the gene showed mice from the Ahr(b2) and Ahr(d) alleles in separate branches while mice from the Ahr(b1) and Ahr(b3) alleles exhibited a more complex history. Evolutionarily, the AHR protein as a whole appears to be under purifying selective pressure (K(a) : K(s) ratio = 0.237). Despite significant functional constraint in the basic helix-loop-helix and PAS domains, ligand binding is not constrained to the high-affinity allele, which supports further the role of the AHR in development and its importance beyond the adaptive response to environmental toxicants. PMID- 11875371 TI - Glutathione transferase GSTT1, broccoli, and prevalence of colorectal adenomas. PMID- 11875370 TI - Pharmacological properties of the naturally occurring Ala(457)Pro variant of the human norepinephrine transporter. AB - Recently, another research group has reported an almost complete loss of function of the human norepinephrine transporter (hNET) in patients who had orthostatic intolerance and who were heterozygous for a guanine to cytosine exchange, resulting in a hNET Ala(457)Pro variant. To explore the reason for the deficiency in NET function, we compared in detail the pharmacology of the Ala(457)Pro variant with that of the wild-type hNET in COS-7 cells transiently transfected with hNET or Ala(457)Pro cDNA. Compared to the wild-type hNET, the Ala(457)Pro variant exhibited a five-fold higher affinity for cocaine, but a two-fold lower affinity for the NET inhibitor nisoxetine, and an unchanged affinity for the antidepressant desipramine. Plasma membrane expression (measured as Bmax of [3H]nisoxetine binding) of the Ala(457)Pro variant was only 40% of that of the wild-type hNET. The Ala(457)Pro variant showed a six- to 10-fold decrease in affinity for the substrates dopamine and 1-methyl-4-phenylpyridinium (MPP(+)). Compared with the wild-type hNET, the maximum rate (V(max)) of norepinephrine uptake by the Ala(457)Pro variant was slightly reduced, whereas the turnover number (calculated from V(max)/B(max)) was approximately two-fold higher. However, the Ala(457)Pro variant exhibited a 50-fold higher K(m) (i.e. lower apparent affinity) for norepinephrine than the wild-type hNET. Thus, the previously reported loss of function of the Ala(457)Pro variant associated with orthostatic intolerance is only partly due to a reduction in plasma membrane expression of the transporter, and is mainly caused by the pronounced reduction in the apparent affinity of norepinephrine. PMID- 11875372 TI - Relapses after treatment of external genital warts are more frequent in HIV positive patients than in HIV-negative controls. AB - BACKGROUND: Recurrences of cervical lesions associated with human papillomavirus are more frequent in HIV-infected (HIV+) than in HIV- women. Recurrences of external genital warts were investigated in HIV+ patients and HIV- control subjects. GOAL: To compare relapses after treatment of external genital warts between HIV+ and HIV- patients. STUDY DESIGN: At the sexually transmitted disease (STD) center in Brescia, Italy, 1336 patients (241 HIV+ and 1095 HIV-) with external genital warts were examined in the decade 1990 to 1999. Various local treatments were used. RESULTS: Treatments generally triggered recovery from the lesions. The relapses observed up to 1 year after the response, examined by survival analysis, were significantly (P < 0.001) more frequent in the HIV+ (160 cases; 66.4%) than in the HIV- (294 cases; 26.8%) subjects. Multiple relapses observed up to 1 year after treatment occurred in 69 of 241 HIV+ patients, as compared with 14 of 1095 HIV- control subjects (P < 0.001). CONCLUSION: According to the study findings, HIV infection can be considered a risk factor for the development and recurrence of external genital warts. Multiple relapses should drive patients to HIV testing. PMID- 11875373 TI - Prevalence and determinants of sexually transmitted diseases: an analysis of young Jamaican males. AB - BACKGROUND: Jamaican adolescents have high rates of sexually transmitted diseases (STDs). GOALS: Since the sexual behaviors that put an individual at risk for HIV are the same as for other STDs, the prevalence and determinants of STD symptoms among a sample of young Jamaican males were examined. STUDY DESIGN: As part of the 1997 Reproductive Health Survey, male adolescents and young adults in Jamaica were surveyed about symptoms of STDs and related sex behaviors. RESULTS: Overall, 9% of the sample reported symptoms of STDs in the year before the interview. Rates of high-risk sexual behaviors were high. Logistic regression analyses indicated that being older and having multiple sex partners were associated with having symptoms of STDs. CONCLUSIONS: Prevention programs should recognize that various factors can increase the risk of contracting and transmitting STDs, including HIV. Interventions should be targeted to those with high-risk behaviors that are conducive to continued participation in high-risk sexual behaviors. PMID- 11875374 TI - Sex partner concurrency: measurement, prevalence, and correlates among urban 18 39-year-olds. AB - BACKGROUND: Sex partner concurrency probably accelerates the spread of sexually transmitted disease (STD) and HIV, yet few data exist on population prevalence or correlates. GOAL: The goal of the study was to compare definitions and estimate the frequency of concurrent partnerships and to identify individual and partnership correlates of con-currency. STUDY DESIGN: A random-digit-dialing survey (n = 637) was performed to collect demographic information, sexual history and history of STD, and partnership characteristics. RESULTS: Men reported concurrency more frequently than women. For men, lifetime partners (odds ratio [OR], 1.15 per partner; 95% CI, 1.07-1.23), a night in jail (OR, 1.99; 95% CI, 1.03-3.82), and same sex partners (OR, 1.88; 95% CI, 0.92-3.84) were associated with concurrency. Important factors for women were first coitus before age 16 (OR, 2.90; 95% CI, 1.38-6.10), lifetime partners (OR, 1.09 per partner; 95% CI, 1.01-1.16), and STD diagnoses during relationship (OR, 3.53; 95% CI, 1.55-8.05). Partnership characteristics associated with concurrency included lifetime partners (OR, 1.09; 95% CI, 1.05-1.14), race discordance (OR, 1.72; 95% CI, 1.14 2.59), married/living together (OR, 0.60; 95% CI, 0.36-0.98), night in jail (OR, 2.04; 95% CI, 1.32-3.17), partnership duration of >6 months (OR, 2.43; 95% CI, 1.41-4.19), and STD diagnoses during relationship (OR, 2.68; 95% CI, 1.42-5.07). CONCLUSIONS: Concurrency was independently associated with individual STD risk. Sex differences may reflect true behavioral differences or differential reporting. PMID- 11875376 TI - Screening for syphilis in arrestees: usefulness for community-wide syphilis surveillance and control. AB - BACKGROUND: Syphilis screening of jail arrestees has been promoted as an effective method for both disease control and surveillance. GOALS: To evaluate the yield of the East Baton Rouge Parish Jail screening program in detecting previously undiagnosed syphilis, to evaluate the program as a means for monitoring community syphilis rates, and to characterize arrestees at greatest risk for syphilis infection. STUDY DESIGN: From July 1994 to December 1998, arrestees brought to the East Baton Rouge Parish Jail were screened for syphilis. Annual early syphilis prevalence in screened arrestees was calculated and compared with the annual period prevalence of early syphilis in the general population of East Baton Rouge Parish, as reported by laboratories and health providers. A case-control study of cases detected at the jail from 1995 to 1997 and contemporary controls was conducted. RESULTS: A total of 50,941 arrestees were booked into the East Baton Rouge Parish Jail, of whom 38,573 (76%) were screened for syphilis. Of the 38,573 arrestees screened, 494 (1.3%) were diagnosed with untreated syphilis. Of these, 299 (61%) were treated for syphilis before release. The estimated prevalence of early syphilis in arrestees decreased by 68% during the study period, from 0.79% in 1994 to 0.25% in 1998. During this time, the East Baton Rouge Parish community rates decreased by 79%, from 150 cases per 100,000 to 31 cases per 100,000. In female arrestees, a booking charge of prostitution was associated with syphilis (odds ratio [OR] 7.0; 95% CI, 1.5, 39.3). In male arrestees, a booking charge of felony theft was associated with syphilis (OR 4.8; 95% CI, 1.8, 13.8). However, only 15 (12%) of the early syphilis cases would have been detected if screening had been based on the booking charges found to be associated with syphilis in this study. CONCLUSIONS: Routine syphilis screening and treatment in jail settings is feasible and identifies many persons with syphilis. Monitoring of syphilis prevalence among arrestees is a useful method for monitoring community prevalence of syphilis. Analysis of booking charges may be useful for determining factors associated with syphilis infection, but not for developing screening criteria. PMID- 11875375 TI - Older partners not associated with recurrence among female teenagers infected with Chlamydia trachomatis. AB - BACKGROUND: Chlamydia trachomatis-infected female teenagers with older partners may be less likely to discuss the infection with their partner(s) and to use condoms and therefore may be more likely to get reinfected. GOAL: To determine if C trachomatis-infected female teenagers with older partners were more likely to be reinfected than those with same-aged partners. STUDY DESIGN: Females aged 14 years to 18 years who had uncomplicated chlamydial infection, were nonpregnant, attended clinics in five United States cities from June 1995 to May 1997, completed treatment, and resumed sexual activity were observed at 1 and 4 months for interim history and retesting. RESULTS: Of 225 women studied, 73.3% were black, 34.5% had at least one partner who was 3 or more years older during follow up, 51.6% reported using a condom at the last sex act with all partners, 13.8% had a recurrent infection, and 47.4% reported they were certain that all of their baseline partners were treated. Partner age was not associated with condom use, certainty of partners' taking medication, or recurrent infections after adjustment for visit. CONCLUSIONS: Older partners, accounting for approximately one third of all partners, did not increase the risk of reinfection. Given the high risk for recurrence, follow-up testing and enhanced efforts to ensure partner treatment are appropriate for all young women with chlamydial infections. PMID- 11875377 TI - The impact of syphilis, HIV-1, and HIV-2 on pregnancy outcome in Bissau, Guinea Bissau. AB - BACKGROUND: Syphilis remains a major cause of fetal loss and neonatal mortality in sub-Saharan Africa. Guinea-Bissau is the epicenter of the HIV-2 epidemic, and little is known about the impact of HIV-2 on pregnancy. GOAL: To understand better the impact of maternal syphilis and HIV-2 on the outcomes of pregnancy in Bissau, Guinea-Bissau. STUDY DESIGN: Using a case-control design, maternal syphilis and HIV-2 seropositive results were examined in relation to adverse outcomes of pregnancy. From June 1997 to April 1998, women presenting to the Simao Mendes hospital of Bissau for a delivery or a spontaneous abortion were invited to participate in the study, and 1341 women were enrolled. The 743 control subjects were women who had delivered a term neonate with a birthweight greater than 2500 g that survived the first 28 days of life. The cases were classified into five groups of mothers according to the outcome of pregnancy: stillbirths (n = 185), spontaneous abortions (n = 89), premature deliveries (n = 256), small-for-gestation-age babies (n = 55), and neonatal deaths (n = 13). RESULTS: Among the control subjects, the prevalences of serologic syphilis and HIV infection were 3.9% and 7.9%, respectively. Positive syphilis serology results together with a rapid plasma reagin titer of 1:16 or more were associated with delivery of a stillborn (adjusted odds ratio [AOR], 6.05) and premature delivery (AOR, 2.98). In Bissau, the population-attributable risk fraction of syphilis was 7.2% (95% CI, 2.2-11.9%) for stillbirths and only 2.4% (95% CI, 0 5.8%) for premature deliveries. Spontaneous abortions, delivery of a small-for gestation-age baby, and neonatal deaths were not associated with positive syphilis serology results. None of these pregnancy outcomes was significantly associated with HIV-2 infection. CONCLUSIONS: The effect of syphilis on the outcome of pregnancy in West Africa is similar to that reported from other parts of Africa. However, because the prevalence of positive syphilis serology results is relatively low, the impact of screening for syphilis on the outcome of pregnancy is likely to be relatively modest. The findings from this study confirm the absence of association between HIV-2 and an adverse pregnancy outcome. PMID- 11875378 TI - Dual protection against unintended pregnancy and sexually transmitted infections: what is the best contraceptive approach? AB - In the midst of the global epidemics of both unintended pregnancy and sexually transmitted infection, contraceptive options that provide dual protection are ideal. However, those contraceptives with the best record of preventing pregnancy under typical use conditions (sterilization, hormonal methods, intrauterine devices) provide little if any protection against sexually transmitted infection. Alternatively, barrier contraceptive methods (specifically, condoms), which can reduce risks of many sexually transmitted infections, are associated with relatively higher pregnancy rates for most users than other contraceptives. This situation has produced a dilemma for those wishing to promote dual protection: whether to advocate use of two methods (one primarily to prevent pregnancy and the other primarily to prevent infections) or whether to emphasize use of condoms for both purposes. Data comparing these two approaches are limited and often contradictory. We discuss the underlying concepts of exposure to both pregnancy and infection, provide a broad overview of the effectiveness of contraceptive methods against these two conditions, present approaches to optimize dual protection, and propose several new directions for necessary research. In the absence of evidence-based recommendations, we believe clinicians should assist clients in assessing their likelihood of exposure to infection, either by prevalence of sexually transmitted infection in the community or by the specific risk factors of the client. If exposure is likely, particularly to the more serious infections such as human immunodeficiency virus, the one-method approach should be given greater weight. However, in settings where unintended pregnancy is the greater concern, emphasizing the two-methods approach as a first option may be appropriate. PMID- 11875379 TI - Herpes simplex virus type 2 seropositivity among urban adults in Africa: results from two cross-sectional surveys in Addis Ababa, Ethiopia. AB - BACKGROUND: Although several surveys investigating the epidemiology of herpes simplex virus type 2 (HSV-2) infection using type-specific immunologic assays have been carried out in Africa, none has examined the risk factors for HSV-2 infection in a representative sample from an urban adult population. GOALS: To estimate the prevalence of HSV-2 infection in the adult population of Addis Ababa, Ethiopia, and to identify risk factors for HSV-2 infection. STUDY DESIGN: Two cross-sectional surveys, one community-based (June to September 1996, n = 506) and one factory-based (February to November 1997, n = 657), were conducted. Samples were tested for HSV-2 immunoglobulin G antibodies using type-specific enzyme-linked immunoassays (ELISA). RESULTS: In the community-based survey, HSV-2 prevalence increased with age until 25 years, then leveled off at 50% in both genders. The same independent predictors of HSV-2 infection were identified in both genders: older age, higher lifetime number of sexual partners, positive HIV serology, and positive Treponema pallidum hemagglutination serology. CONCLUSIONS: This study confirmed the high prevalence of HSV-2 infection among adults in an African urban population and its association with HIV infection. Prevention of HSV-2 and other sexually transmitted infections through partner reduction and condom use should be encouraged. PMID- 11875380 TI - Sexual behavior, human papillomavirus type 16 (HPV 16) infection, and HPV 16 seropositivity. AB - BACKGROUND: Sexual behaviors have been linked to seropositivity for human papillomavirus (HPV) but not with the magnitude of the seroreactivity. GOALS: The objective of this analysis was to examine the association of sexual behavior, cervical HPV 16 DNA positivity at enrollment (past) and at diagnosis (current), and other potential determinants with the likelihood and magnitude of HPV 16 seropositivity at diagnosis. STUDY DESIGN: With use of stored specimens from an incidence case-control study at Kaiser Permanente (Portland, OR), women were tested for seroreactivity to HPV 16 by enzyme-linked immunosorbent assay with virus-like particles at diagnosis and were tested for past and concurrent cervical HPV 16 DNA positivity with MY09/MY11 L1 consensus primer PCR. Questionnaire data were used to ascertain past sexual behavior. RESULTS: Increased lifetime number of sex partners (P(Trend) < 0.001), past HPV 16 DNA positivity (odds ratio = 6.9; 95% confidence interval = 1.5-31), and a current cytologic diagnosis (P(Trend) < 0.03) were independently associated with HPV 16 seropositivity. Among the seropositive, only lifetime number of sex partners (P(Trend) < 0.001) and past HPV 16 DNA positivity (P = 0.003) were independently associated with mean signal strength (optical density) in an age-adjusted analysis. Women negative for past and concurrent HPV 16 DNA had a significant trend of increasing optical densities associated with greater numbers of lifetime partners (P(Trend) < 0.001). Conversely, the mean signal strength for those women who were ever HPV 16 DNA-positive during the study did not depend on lifetime numbers of sex partners (P(Trend) = 0.36). CONCLUSIONS: HPV 16 seropositivity is a surrogate for past HPV 16 infection. Circulating levels of antibodies to HPV 16 may reflect recent HPV 16 infection or the frequency of past HPV 16 infection. PMID- 11875381 TI - Assessment of coronary stenoses of graded severity by myocardial contrast echocardiography. AB - BACKGROUND: Myocardial contrast echocardiography (MCE) has potential value in the assessment and quantitation of myocardial perfusion defects. However, the severity of stenosis detectable by MCE and its diagnostic accuracy remain undefined. Thus, we produced coronary stenoses of variable severity and quantified their effect on MCE. METHODS AND RESULTS: Three grades of left anterior descending (LAD) obstructions were produced in 7 open-chest swine. The stenoses were nonflow-limiting at rest, but decreased coronary hyperemia by 31.3% +/- 4.7%, 69.9% +/- 5.3% and 98.9% +/- 1.1%, respectively. Regional myocardial blood flow (RBF) was measured with fluorescent microspheres and was expressed as the ratio of LAD and control (LCx) beds. MCE was performed with 0.3 mg/kg intravenous AF0150 during ECG-gated harmonic imaging in short-axis view. Background-subtracted peak intensity (PI) was expressed as the ratio of LAD/LCx beds. Both RBF and PI ratios progressively decreased with increasing grades of stenosis. MCE showed a significant correlation with RBF (r = 0.74; P <.0001). Ratios of both PI and RBF differed significantly from baseline when coronary hyperemia was reduced more than 50%. An LAD/LCx ratio less than 0.6 by MCE yielded 61% and 83% sensitivity and 85% and 76% specificity with stenosis that reduced coronary hyperemia more than 50% and more than 75%, respectively. CONCLUSION: MCE with intravenous AF0150 during vasodilation correctly depicted the progressive reduction of flow ratios produced by graded coronary stenoses. A significant reduction of PI ratio was observed with stenosis causing more than 50% reduction of coronary hyperemia. An MCE ratio in stenosed/control beds could be selected, which exhibited good sensitivity and specificity in the identification of coronary stenosis. PMID- 11875382 TI - Quantitative assessment of left ventricular perfusion defects using real-time three-dimensional myocardial contrast echocardiography. AB - Quantitative assessment of perfusion defects with myocardial contrast echocardiography can be a valuable tool in the evaluation of patients with coronary artery disease. However, the use of 2-dimensional echocardiography for this purpose is limited to a restricted number of imaging planes. Real-time 3 dimensional echocardiography (RT3D) is a novel technique that provides instantaneous volumetric images. The aim of this study was to validate the use of RT3D for the quantitative assessment of myocardial perfusion defects in a model of acute coronary occlusion. To this end, 20 sheep underwent acute ligation of the left anterior descending (n = 14) or the posterior branch of the circumflex (n = 6) artery under general anesthesia. The RT3D images were obtained after left atrial injection of the contrast agent EchoGen (perflenapent emulsion; 0.8-1 mL). Evans blue dye was injected into the occluded coronary artery for subsequent anatomic identification of underperfused myocardium. The mass of the entire left ventricle and of the underperfused myocardial region were measured after death. Blinded off-line calculation of left ventricular (LV) mass and perfusion-defect mass from RT3D images were performed using an interactive aided-manual tracing technique. Total LV mass ranged from 68 to 141 g (mean plus minus SD: 92 +/- 24 g). The mass of the perfusion defect ranged from 0 to 43 g (mean +/- SD: 16 +/- 9 g) or 0 to 36% of total LV mass (mean +/- SD: 18% +/- 9%). The RT3D estimation of total LV mass strongly correlated with the anatomic measurement (r = 0.91; y = 2.54 + 1.04x; standard error of the estimate [SEE] = 11.9 g). The RT3D calculation of the mass of underperfused myocardium also strongly correlated with the anatomic measurement, both in absolute terms (r = 0.96; y = 2.01 + 0.87x; SEE = 2.2 g) and when expressed as percentage of total LV mass (r = 0.96; y = 0.11 + 1.02x; SEE = 2.8%). Hence, RT3D with myocardial contrast opacification accurately predicts the amount of underperfused myocardium in an animal model of acute coronary occlusion. This technique may therefore be useful for the quantitative assessment of myocardial perfusion defects in patients with coronary artery disease. PMID- 11875383 TI - Ultrasound-mediated transfection of canine myocardium by intravenous administration of cationic microbubble-linked plasmid DNA. AB - We tested the hypothesis that targeted disruption of cationic microbubble-linked plasmid DNA, using diagnostic ultrasound, may aid transfection of large animal myocardium. Plasmid DNA encoding for CAT (pCAT, chloramphenicol acetyltransferase) was bound to a novel cationic microbubble containing MRX-225 for intravenous administration, and 16 dogs in 4 groups variously received this conjugate or plasmid only, or were exposed to ultrasound. Histochemical staining and enzyme-linked immunosorbent assay analysis showed CAT activity in the myocardium of only those animals that received microbubble-linked DNA and were exposed to ultrasound. Thus, disruption of cationic-linked, low-dose plasmid systems by diagnostic ultrasound may facilitate transfection of large animal hearts. PMID- 11875384 TI - Hydrodynamics of color M-mode Doppler flow wave propagation velocity V(p): a computer study. AB - This study was designed to show the hydrodynamic mechanism of left ventricular (LV) flow wave propagation and to relate this propagated velocity to 2 dimensional (2D) color and color M-mode Doppler echocardiograms. A computer model is developed describing 3-dimensional axisymmetrical LV filling flow. The unsteady Navier-Stokes flow equations are solved in an LV truncated ellipsoid geometry with moving LV walls, including relaxation and compliance of the wall. The computed results confirm both intraventricular flow and pressure patterns during filling. Vortices are formed during the acceleration phases of the early and atrial filling waves. During the deceleration phases, the vortices are amplified and convected into the ventricle. The vortices are recognized on the derived 2D color echocardiograms as in vivo. The propagation of this vortex determines the propagation of the maximum velocity observed in the color M-mode Doppler echocardiogram. For pseudonormal filling of the left ventricle, the LV flow wave propagation velocity decreases. PMID- 11875385 TI - Quantification of left ventricular diastolic pressure-volume relations during routine cardiac catheterization by two-dimensional digital echo quantification and left ventricular micromanometer. AB - BACKGROUND: Currently there is no simple clinical method for quantifying the left ventricular (LV) diastolic pressure-volume relation. Echocardiographic-automated endocardial border detection, however, may be combined with LV micromanometer to construct LV pressure-volume loops. We investigated the feasibility of on-line display and sampling of LV pressure-volume loops by such an approach. For this purpose we used a new echocardiographic digital echo quantification (DEQ) method in combination with LV pressures on-line and in real-time. METHODS: Eighteen patients were screened by conventional echocardiography and DEQ. Ten of the patients with high quality images were included in the study. Left ventricular pressures and volumes were recorded simultaneously and were displayed on-line as pressure-volume loops. Changes in LV volume were induced by intravenous saline. Left ventricular chamber compliance was estimated as change in volume divided by change in pressure from minimum diastolic pressure to end-diastolic pressure (average LV chamber compliance). RESULTS: Left ventricular pressure-volume loops were displayed on-line during the examination. When compared with the Simpson's method, DEQ underestimated end-diastolic volume (EDV) by 35% and overestimated end-systolic volume (ESV) by 14%. Beat-to-beat variability for ESV and EDV were 7.4% +/- 0.8% and 7.2% +/- 0.7 %, respectively. Volume loading increased LV end diastolic pressure (LVEDP) from 14.0 +/- 1.6 to 24.7 +/- 2.0 mm Hg (P <.05) and EDV from 79 +/- 10 to 85 +/- 11 mL (NS), and decreased LV chamber compliance from 4.0 +/- 0.7 to 2.0 +/- 0.3 mL/mm Hg (P <.05). CONCLUSION: The current study demonstrates that LV pressure-volume loops can be displayed and evaluated in real time during routine cardiac catheterization. This may represent a clinically useful method for identifying patients with reduced chamber compliance. The underestimation of the volumes by DEQ compared with the Simpson's method suggests that further refinements should be performed to improve the endocardial border detection algorithm. PMID- 11875386 TI - Quantitative assessment of regurgitant flow with total digital three-dimensional reconstruction of color Doppler flow in the convergent region: in vitro validation. AB - BACKGROUND: This study was designed to develop and test a total digital 3 dimensional (3D) color flow map reconstruction for proximal isovelocity surface area (PISA) measurement in the convergent region. METHODS: Asymmetric flow convergent velocity field was created in an in vitro pulsatile model of mitral regurgitation. Image files stored in the echocardiographic scanner memory were digitally transferred to a computer workstation, and custom software decoded the file format, extracted velocity information, and generated 3D flow images automatically. PISA and volume flow rate were calculated without geometric assumption. For comparison, regurgitant volume was also calculated, using continuous wave Doppler, 2-dimensional (2D), and M-mode color flow Doppler with the hemispheric approach. RESULTS: Flows from 3D digital velocity profiles showed a closed, excellent relation with actual flow rates, especially for instantaneous flow rate. Regurgitant volume calculated with the 3D method underestimated the actual flow rate by 2.6%, whereas 2D and the M-mode method show greater underestimation (44.2% and 32.1%, respectively). CONCLUSION: Our 3D reconstruction of color flow Doppler images gives more exact information of the flow convergent zone, especially in complex geometric flow fields. Its total digital velocity process allows accurate measurement of convergent surface area and improves quantitation of valvular regurgitation. PMID- 11875387 TI - Evaluation of valvular regurgitation severity using digital acquisition of echocardiographic images. AB - BACKGROUND: Digital acquisition is a technique for storing echocardiographic data that offers advantages over conventional videotape (VT); however, limited information is available on its accuracy for the evaluation of valvular regurgitation. METHODS: We evaluated 102 patients with at least 1 regurgitant lesion. Data were obtained on VT and in 1 cardiac cycle stored digitally (1C). To assess for incremental improvement with acquisition of multiple cycles, digital images were also acquired with 2 (2C) or 3 cardiac cycles (3C). Both digital and VT images were graded for regurgitant severity as absent, trivial, mild, moderate, or severe. Kappa statistics were used to assess agreement. RESULTS: A total of 171 valvular regurgitant lesions (mild or greater) were evaluated. The overall agreement between 1C and VT images was kappa = 0.61. With multiple cycle acquisition, there was no improvement in agreement (kappa = 0.56 and 0.57 for 2C and 3C, respectively). When subgrouped, the level of agreement between 1C and VT was slightly lower for the aortic valve than for the mitral or tricuspid valves (kappa = 0.49, 0.63, 0.64, respectively). CONCLUSION: The 1C technique has substantial agreement and correlation with standard VT for the evaluation of regurgitant lesions with the use of color flow Doppler. The acquisition of multiple cardiac cycles does not provide incremental improvement over single beat acquisition. PMID- 11875388 TI - Ventricular septal flattening at end systole falsely predicts right ventricular hypertension in patients with ostium primum atrial septal defects. AB - PURPOSE: To assess the reliability of ventricular septal position in predicting elevated right ventricular pressure (RVP) in patients with ostium primum atrial septal defects (ASD 1). METHODS: Echocardiograms of 4 groups were retrospectively analyzed: Patients with ASD 1 and low RVP, patients with ASD 1 and high RVP, and 2 age-matched control groups: one with isolated ostium secundum atrial septal defects (ASD 2), and 1 with normal cardiac findings. End-systolic left ventricular sectional diameters along the midmitral diameter (D1) and a diameter orthogonal to it (D2) were measured off-line by a blinded observer. The ratio D2/D1, the eccentricity index (EI), was calculated; a higher index represents greater septal flattening. RESULTS: The mean EI in the ASD 1 with low RVP group was significantly higher than both the group with ASD 2 and the healthy control group. The mean EI of the ASD 1 group with high RVP was significantly higher than the mean EI of the ASD 1 group with low RVP, although there was a poor correlation between EI and RVP in this group, r = 0.54. CONCLUSION: The ventricular septum is flatter in the ASD 1 patients with low RVP than in an age matched control group with ASD 2 and compared with an age-matched control group of healthy subjects, giving a false impression of elevated RVP in the ASD 1 group. Although the mean EI is significantly higher in the ASD 1 group with high RVP than in the group with low RVP, there is a poor correlation between EI and RVP, which limits the reliability of this index. PMID- 11875389 TI - Noninvasive assessment of great cardiac vein flow by Doppler echocardiography: a validation study. AB - The objectives of this study were (1) to compare great cardiac vein (GCV) flow velocity detected by pulsed Doppler echocardiography (PDE) with Doppler guide wire (DGW) in the experimental setting and (2) to clarify whether transthoracic Doppler echocardiography (TTDE) can detect GCV flow in humans. Using opened-chest dogs, we detected GCV flow by PDE under the guidance of color flow Doppler mapping. GCV flow velocity was recorded by PDE and DGW, simultaneously. In 23 volunteers, GCV flow velocity was measured by TTDE. In the experimental setting, the prominent systolic flow wave of the GCV was obtained in PDE and DGW. There were good agreements between PDE and DGW for the measurements of GCV flow velocity (peak velocity: r = 0.98, y = 1.12chi-5.9; time velocity integral: r = 0.97, y = 1.10chi-0.71). In the human subjects, clear envelopes of GCV flow velocity were obtained in 21 (91%) of 23 subjects with the use of TTDE. PMID- 11875390 TI - Detection of septal coronary collaterals by color flow Doppler mapping is a marker for anomalous origin of a coronary artery from the pulmonary artery. AB - Between August 1991 and September 2000, 15 patients received a diagnosis of the anomalous origin of the coronary artery from the pulmonary artery, at the Children's Hospital of Wisconsin. All were evaluated initially by transthoracic echocardiography, with subsequent diagnosis confirmation at cardiac catheterization and/or surgery. Seven of the 15 patients were referred in infancy (mean age 4.3 months) with symptoms of congestive heart failure. The remaining 8 patients were older (mean age 7.0 years) at the time of diagnosis, and 7 of those 8 patients were clinically asymptomatic and were referred for evaluation of a heart murmur and/or cardiomegaly on chest radiograph. One older patient, previously healthy, was referred at age 18 for an episode of sudden death while playing basketball. All the older asymptomatic patients had echocardiographic detection of multiple unusual color flow Doppler signals within the ventricular septum, believed to represent septal coronary collaterals, which raised suspicion of a coronary artery abnormality and led to more detailed imaging of the coronary artery anatomy. In the younger infants with congestive heart failure, septal coronary collaterals were less frequent, but did aid in the diagnosis of an anomalous coronary artery when present. PMID- 11875391 TI - Anomalous origin of right coronary artery above the sinus of Valsalva: observation by transthoracic echocardiography. AB - We describe a 60-year-old woman with an anomalous right coronary artery originating from the tubular portion of the ascending aorta. The anomaly was diagnosed incidentally by transthoracic echocardiography. PMID- 11875392 TI - Nonobstructive membranes of the left atrial appendage cavity: report of three cases. AB - A membranous structure causing functional stenosis at the mouth of the left atrial appendage (LAA) has been reported. In this study we describe the presence of nonobstructive membranes traversing the cavity of the LAA found incidentally on transesophageal echocardiography (TEE). PMID- 11875393 TI - Role of transesophageal echocardiography in diagnosis and management of cardiac hydatid cyst: report of three cases and review of the literature. AB - Although diagnostic value of transthoracic echocardiography in cardiac echinococcus is well established, the role of transesophageal echocardiography in both the diagnosis and the management of this entity is not well known. We present 3 unusual cases of cardiac hydatid cyst in which transesophageal echocardiography was used. A review of the literature on the subject is also presented. PMID- 11875394 TI - Recommendations for a standardized report for adult transthoracic echocardiography: a report from the American Society of Echocardiography's Nomenclature and Standards Committee and Task Force for a Standardized Echocardiography Report. PMID- 11875395 TI - Jet eccentricity: a misleading source of agreement between Doppler/catheter pressure gradients in aortic stenosis. PMID- 11875397 TI - The AID enzyme induces class switch recombination in fibroblasts. AB - The switch of the immunoglobulin isotype from IgM to IgG, IgE or IgA is mediated by class switch recombination (CSR). CSR changes the immunoglobulin heavy chain constant region (CH) gene from Cmu to one of the other CH genes. Somatic hypermutation introduces massive numbers of point mutations in the immunoglobulin variable (V) region gene, giving rise to immunoglobulin with higher affinity. Activation-induced cytidine deaminase (AID), a putative RNA-editing cytidine deaminase, is expressed strictly in activated B cells and is indispensable in both CSR and somatic hypermutation. But the exact function of AID is unknown. Here we show that ectopic expression of AID induces CSR in an artificial switch construct in fibroblasts at a level comparable to that in stimulated B cells. Sequences around recombination junctions in the artificial substrate have features similar to endogenous CSR junctions. Furthermore, AID-induced CSR in fibroblasts is dependent on transcription of the target S region, as shown in endogenous CSR in B cells. The results show that AID is the only B-cell-specific factor required for initiation of the CSR reaction in the activated locus. PMID- 11875398 TI - Diurnal modulation of pacemaker potentials and calcium current in the mammalian circadian clock. AB - The central biological clock of the mammalian brain is located in the suprachiasmatic nucleus. This hypothalamic region contains neurons that generate a circadian rhythm on a single-cell basis. Clock cells transmit their circadian timing signals to other brain areas by diurnal modulation of their spontaneous firing rate. The intracellular mechanism underlying rhythm generation is thought to consist of one or more self-regulating molecular loops, but it is unknown how these loops interact with the plasma membrane to modulate the ionic conductances that regulate firing behaviour. Here we demonstrate a diurnal modulation of Ca2+ current in suprachiasmatic neurons. This current strongly contributes to the generation of spontaneous oscillations in membrane potential, which occur selectively during daytime and are tightly coupled to spike generation. Thus, day night modulation of Ca2+ current is a central step in transducing the intracellular cycling of molecular clocks to the rhythm in spontaneous firing rate. PMID- 11875399 TI - Will these pigs ever fly? PMID- 11875400 TI - I'll be back. PMID- 11875401 TI - UNEP "buys support for Cartagena," say critics. PMID- 11875403 TI - Panels jockey for position in US debate on cloning. PMID- 11875406 TI - Organic farmers sue GMO producers. PMID- 11875408 TI - Nontransgenic crops from transgenic plants. PMID- 11875409 TI - How to publish DNA sequences with copyright protection. PMID- 11875410 TI - Great companies, bad stocks. PMID- 11875412 TI - Real options for biotechnology valuation. PMID- 11875414 TI - ES cell guidelines in Sweden. PMID- 11875415 TI - Germany permits import of ES cells. PMID- 11875416 TI - A perspective on protein microarrays. PMID- 11875417 TI - Knocking out xenograft rejection. PMID- 11875418 TI - A solid base for assaying protein kinase activity. PMID- 11875419 TI - Hitting the sweet spot. PMID- 11875420 TI - Guided genes for tumor warfare. PMID- 11875421 TI - The genesis of embryonic stem cells. PMID- 11875422 TI - Spiderless spider webs. PMID- 11875424 TI - Metabolic control analysis in drug discovery and disease. AB - Metabolic control analysis (MCA) provides a quantitative description of substrate flux in response to changes in system parameters of complex enzyme systems. Medical applications of the approach include the following: understanding the threshold effect in the manifestation of metabolic diseases; investigating the gene dose effect of aneuploidy in inducing phenotypic transformation in cancer; correlating the contributions of individual genes and phenotypic characteristics in metabolic disease (e.g., diabetes); identifying candidate enzymes in pathways suitable as targets for cancer therapy; and elucidating the function of "silent" genes by identifying metabolic features shared with genes of known pathways. MCA complements current studies of genomics and proteomics, providing a link between biochemistry and functional genomics that relates the expression of genes and gene products to cellular biochemical and physiological events. Thus, it is an important tool for the study of genotype-phenotype correlations. It allows genes to be ranked according to their importance in controlling and regulating cellular metabolic networks. We can expect that MCA will have an increasing impact on the choice of targets for intervention in drug discovery. PMID- 11875425 TI - Targeted disruption of the alpha1,3-galactosyltransferase gene in cloned pigs. AB - Galactose-alpha1,3-galactose (alpha1,3Gal) is the major xenoantigen causing hyperacute rejection in pig-to-human xenotransplantation. Disruption of the gene encoding pig alpha1,3-galactosyltransferase (alpha1,3GT) by homologous recombination is a means to completely remove the alpha1,3Gal epitopes from xenografts. Here we report the disruption of one allele of the pig alpha1,3GT gene in both male and female porcine primary fetal fibroblasts. Targeting was confirmed in 17 colonies by Southern blot analysis, and 7 of them were used for nuclear transfer. Using cells from one colony, we produced six cloned female piglets, of which five were of normal weight and apparently healthy. Southern blot analysis confirmed that these five piglets contain one disrupted pig alpha1,3GT allele. PMID- 11875426 TI - Tumor antigen-specific induction of transcriptionally targeted retroviral vectors from chimeric immune receptor-modified T cells. AB - High-level systemic delivery of viral vectors to tumors has proved problematic as a result of immune neutralization, nonspecific adhesion, and clearance of circulating viral particles. Some cell types localize to tumors in response to particular biological properties associated with tumor growth. Their use to deliver viral vectors to tumors would allow precious viral stocks to be protected until they can be released at high local concentrations. Here, we describe a mechanism by which retroviral vector production by T cells can be regulated by a tumor-specific trigger through engagement of a chimeric immune receptor (CIR) with its target antigen. The virus that is released from the T cells can also be transcriptionally targeted. Finally, we show that it is possible to use vector loaded, antigen-triggered human T cells as therapeutic, tumor-specific vector delivery cells in models of both local intratumoral and systemic delivery to both lung and liver metastases. This strategy incorporates multiple levels of targeting into the delivery system at the stages of surface targeting, viral production, and gene expression. PMID- 11875427 TI - Enhancement of the antitumor activity of interleukin-12 by targeted delivery to neovasculature. AB - Interleukin-12 (IL-12) is a heterodimeric cytokine with potent immunostimulatory activity and anti-angiogenic properties. Its clinical applications are limited, however, by severe side-effects. Here we report that an IL-12 fusion protein, consisting of IL-12 fused to a human antibody fragment specific to the oncofetal ED-B domain of fibronectin, markedly enhances the antitumor activity of this cytokine, as demonstrated in a mouse lung-metastasis model and in two models of mice bearing different aggressive murine tumors. The residual small tumor masses seen in the treated mice were infiltrated with lymphocytes, macrophages, and natural killer cells and had elevated interferon gamma (IFN-gamma). These results are of therapeutic relevance as the ED-B domain of fibronectin, a naturally occurring marker of angiogenesis identical in mouse and man, is expressed in the majority of aggressive solid tumors but is not detectable in normal vessels and tissues. PMID- 11875428 TI - Peptide chips for the quantitative evaluation of protein kinase activity. AB - Peptide chips are an emerging technology that could replace many of the bioanalytical methods currently used in drug discovery, diagnostics, and cell biology. Despite the promise of these chips, their development for quantitative assays has been limited by several factors, including a lack of well-defined surface chemistries to immobilize peptides, the heterogeneous presentation of immobilized ligands, and nonspecific adsorption of protein to the substrate. This paper describes a peptide chip that overcomes these limitations, and demonstrates its utility in activity assays of the nonreceptor tyrosine kinase c-Src. The chip was prepared by the Diels-Alder-mediated immobilization of the kinase substrate AcIYGEFKKKC-NH(2) on a self-assembled monolayer of alkanethiolates on gold. Phosphorylation of the immobilized peptides was characterized by surface plasmon resonance, fluorescence, and phosphorimaging. Three inhibitors of the enzyme were quantitatively evaluated in an array format on a single, homogeneous substrate. PMID- 11875429 TI - Carbohydrate microarrays for the recognition of cross-reactive molecular markers of microbes and host cells. AB - We describe here the development of a carbohydrate-based microarray to extend the scope of biomedical research on carbohydrate-mediated molecular recognition and anti-infection responses. We have demonstrated that microbial polysaccharides can be immobilized on a surface-modified glass slide without chemical conjugation. With this procedure, a large repertoire of microbial antigens (approximately 20,000 spots) can be patterned on a single micro-glass slide, reaching the capacity to include most common pathogens. Glycoconjugates of different structural characteristics are shown here to be applicable for microarray fabrication, extending the repertoires of diversity and complexity of carbohydrate microarrays. The printed microarrays can be air-dried and stably stored at room temperature for long periods of time. In addition, the system is highly sensitive, allowing simultaneous detection of a broad spectrum of antibody specificities with as little as a few microliters of serum specimen. Finally, the potential of carbohydrate microarrays is demonstrated by the discovery of previously undescribed cellular markers, Dex-Ids. PMID- 11875430 TI - An electrical probe of protein-DNA interactions on DNA-modified surfaces. AB - DNA charge transport chemistry is found to provide a sensitive method for probing protein-dependent changes in DNA structure and enzymatic reactions. Here we describe the development of an electrochemical assay of protein binding to DNA modified electrodes based upon the detection of associated perturbations in DNA base stacking. Gold electrode surfaces that were modified with loosely packed DNA duplexes, covalently crosslinked to a redox-active intercalator and containing the binding site of the test protein, were constructed. Charge transport through DNA as a function of protein binding was then assayed. Substantial attenuation in current is seen in the presence of the base-flipping enzymes HhaI methylase and uracil DNA glycosylase, as well as with TATA-binding protein. When restriction endonuclease PvuII (R.PvuII) binds to its methylated target, little base-stacking perturbation occurs and little diminution in current flow is observed. Importantly, the kinetics of restriction by R.PvuII of its nonmethylated target is also easily monitored electrochemically. This approach should be generally applicable to assaying protein--DNA interactions and reactions on surfaces. PMID- 11875431 TI - Fluorescent indicators for imaging protein phosphorylation in single living cells. AB - To visualize signal transduction based on protein phosphorylation in living cells, we have developed genetically encoded fluorescent indicators, named phocuses. Two different color mutants of green fluorescent protein (GFP) were joined by a tandem fusion domain composed of a substrate domain for the protein kinase of interest, a flexible linker sequence, and a phosphorylation recognition domain that binds with the phosphorylated substrate domain. Intramolecular interaction of the substrate domain and the adjacent phosphorylation recognition domain within a phocus was dependent upon phosphorylation of the substrate domain by protein kinase, which influenced the efficiency of fluorescence resonance energy transfer (FRET) between the GFPs within a phocus. In the present study, we employed phocuses composed of insulin signaling proteins to visualize protein phosphorylation by the insulin receptor. This method may provide a general approach for studying the dynamics of protein phosphorylation-based signal transduction in living cells. PMID- 11875432 TI - Production of a freeze-thaw-stable potato starch by antisense inhibition of three starch synthase genes. AB - The use of unmodified starches in frozen foods is severely limited by the undesirable textural changes that occur after freezing and thawing. Retrogradation of glucan chains leads to syneresis, a separation of the starch gel and water phases. Stabilization of the starch structure is normally achieved by chemical modification to prevent these changes from occurring. We have now created a freeze-thaw-stable potato starch by alteration of starch composition and structure by genetic modification. An amylose-free starch with short-chain amylopectin was produced by simultaneous antisense downregulation of three starch synthase genes. This starch is extremely freeze-thaw-stable and shows no syneresis even after five freeze-thaw cycles. The use of this starch has potential for environmental and consumer benefits because its production requires no chemical modification. PMID- 11875433 TI - Phosphoproteome analysis by mass spectrometry and its application to Saccharomyces cerevisiae. AB - Protein kinases are coded by more than 2,000 genes and thus constitute the largest single enzyme family in the human genome. Most cellular processes are in fact regulated by the reversible phosphorylation of proteins on serine, threonine, and tyrosine residues. At least 30% of all proteins are thought to contain covalently bound phosphate. Despite the importance and widespread occurrence of this modification, identification of sites of protein phosphorylation is still a challenge, even when performed on highly purified protein. Reported here is methodology that should make it possible to characterize most, if not all, phosphoproteins from a whole-cell lysate in a single experiment. Proteins are digested with trypsin and the resulting peptides are then converted to methyl esters, enriched for phosphopeptides by immobilized metal-affinity chromatography (IMAC), and analyzed by nanoflow HPLC/electrospray ionization mass spectrometry. More than 1,000 phosphopeptides were detected when the methodology was applied to the analysis of a whole-cell lysate from Saccharomyces cerevisiae. A total of 216 peptide sequences defining 383 sites of phosphorylation were determined. Of these, 60 were singly phosphorylated, 145 doubly phosphorylated, and 11 triply phosphorylated. Comparison with the literature revealed that 18 of these sites were previously identified, including the doubly phosphorylated motif pTXpY derived from the activation loop of two mitogen-activated protein (MAP) kinases. We note that the methodology can easily be extended to display and quantify differential expression of phosphoproteins in two different cell systems, and therefore demonstrates an approach for "phosphoprofiling" as a measure of cellular states. PMID- 11875435 TI - Biotechnology patent developments in Europe: an overview. PMID- 11875436 TI - Recent patents in biosensor technology. PMID- 11875439 TI - Effective recruitment strategies for the biotechnology industry. PMID- 11875441 TI - Cancer Research UK. PMID- 11875440 TI - Formins direct Arp2/3-independent actin filament assembly to polarize cell growth in yeast. AB - Formins have been implicated in the regulation of cytoskeletal structure in animals and fungi. Here we show that the formins Bni1 and Bnr1 of budding yeast stimulate the assembly of actin filaments that function as precursors to tropomyosin-stabilized cables that direct polarized cell growth. With loss of formin function, cables disassemble,whereas increased formin activity causes the hyperaccumulation of cable-like filaments. Unlike the assembly of cortical actin patches, cable assembly requires profilin but not the Arp2/3 complex. Thus formins control a distinct pathway for assembling actin filaments that organize the overall polarity of the cell. PMID- 11875443 TI - Calcineurin and skeletal muscle growth. PMID- 11875444 TI - Selector and signalling molecules cooperate in organ patterning. AB - Cell signalling is essential for a plethora of inductive interactions during organogenesis. Surprisingly, only a few different classes of signalling molecules mediate many inductive interactions, and these molecules are used reiteratively during development. This raises the question of how generic signals can trigger tissue-specific responses. Recent studies in Drosophila melanogaster indicate that signalling molecules cooperate with selector genes to specify particular body parts and organ types. Selector and signalling inputs are integrated at the level of cis-regulatory elements, where direct binding of both selector proteins and signal transducers is required to activate tissue-specific enhancer elements of target genes. Such enhancers include autoregulatory enhancers of the selector genes themselves, which drive the refinement of expression patterns of selector genes. PMID- 11875445 TI - Nuclear organization and silencing: putting things in their place. PMID- 11875446 TI - Intervening through interferon. PMID- 11875447 TI - The ins and outs of polycystin-2 as a calcium release channel. PMID- 11875448 TI - Getting hit by SUMO. PMID- 11875449 TI - Many roads lead to the origin. PMID- 11875452 TI - Approaching stalemate? PMID- 11875453 TI - Skewed maturation of virus-specific CTLs? PMID- 11875454 TI - Eradication of poliomyelitis. PMID- 11875455 TI - Requirements for memory maintenance. PMID- 11875456 TI - Gonococci cause immunosuppression by engaging a coinhibitory receptor on T lymphocytes. PMID- 11875457 TI - Antigen receptors rap to integrin receptors. PMID- 11875458 TI - Thymocyte differentiation: it's time to bend a little. PMID- 11875459 TI - T(S) cells and immune tolerance induction: a regulatory renaissance? PMID- 11875461 TI - NF-kappaB at the crossroads of life and death. AB - The choice between life and death is one of the major events in regulation of the immune system. T cells that specifically recognize viral or bacterial antigens are selected to survive and proliferate in response to infection, whereas those that are self-reactive are eliminated via apoptosis. Even the survival of alloreactive T cells requires their proper costimulation and, when infection subsides, the activated T cells are eliminated. A major regulator of such life or death decisions is the transcription factor NF-kappaB. However, NF-kappaB cannot function alone. A variety of mechanisms exist to modulate its activity and thereby affect the ultimate outcome of a cell's fate. PMID- 11875462 TI - Tolerization of dendritic cells by T(S) cells: the crucial role of inhibitory receptors ILT3 and ILT4. AB - Immunoglobulin-like transcript 3 (ILT3) and ILT4 belong to a family of inhibitory receptors expressed by human monocytes and dendritic cells. We show here that CD8+CD28(-) alloantigen-specific T suppressor (TS) cells induce the up-regulation of ILT3 and ILT4 on monocytes and dendritic cells, rendering these antigen presenting cells (APCs) tolerogenic. Tolerogenic APCs show reduced expression of costimulatory molecules and induce antigen-specific unresponsiveness in CD4+ T helper cells. Studies of human heart transplant recipients showed that rejection free patients have circulating TS cells, which induce the up-regulation of ILT3 and ILT4 in donor APCs. These findings demonstrate an important mechanism of immune regulation. PMID- 11875465 TI - Insoluble problem? PMID- 11875466 TI - Integrin-mediated death: an explanation of the integrin-knockout phenotype? PMID- 11875467 TI - Bone marrow does not contribute substantially to endothelial-cell replacement in transplant arteriosclerosis. PMID- 11875468 TI - Bioterrorism becomes one of the hottest US research fields. PMID- 11875469 TI - Hopkins Dean criticizes smallpox research. PMID- 11875470 TI - New investigations into Gulf War syndrome. PMID- 11875471 TI - WHO still struggling to change. PMID- 11875472 TI - Promising drug is victim of bad business. PMID- 11875473 TI - Set back to Alzheimer vaccine studies. PMID- 11875474 TI - Scientists resign over stem-cell study... PMID- 11875475 TI - And reject NIH funds. PMID- 11875476 TI - New study on aging heart. PMID- 11875477 TI - Spanish scientists broke. PMID- 11875478 TI - Royal Society under spotlight. PMID- 11875479 TI - Experts at odds over mammography. PMID- 11875480 TI - Suzanne Cory. PMID- 11875481 TI - Max Ferdinand Perutz 1914--2002. PMID- 11875482 TI - AIDS vaccine models: challenging challenge viruses. PMID- 11875484 TI - Are somatic stem cells pluripotent or lineage-restricted? PMID- 11875485 TI - Are there any normal cloned mammals? PMID- 11875486 TI - Apoptosis and cancer: when BAX is TRAILing away. PMID- 11875487 TI - Telling growth receptors where to go. PMID- 11875488 TI - Epilepsy research gets new guidance. PMID- 11875489 TI - A rendezvous before rejection: where do T cells meet transplant antigens? PMID- 11875490 TI - Shrinking genes for therapy. PMID- 11875492 TI - HIV protease inhibitors are potent anti-angiogenic molecules and promote regression of Kaposi sarcoma. AB - Treatment with HIV-1 protease inhibitors (PI) is associated with a reduced incidence or regression of Kaposi sarcoma (KS). Here we show that systemic administration of the PIs indinavir or saquinavir to nude mice blocks the development and induces regression of angioproliferative KS-like lesions promoted by primary human KS cells, basic fibroblast growth factor (bFGF), or bFGF and vascular endothelial growth factor (VEGF) combined. These PIs also block bFGF or VEGF-induced angiogenesis in the chorioallantoic membrane assay with a potency similar to paclitaxel (Taxol). These effects are mediated by the inhibition of endothelial- and KS-cell invasion and of matrix metalloproteinase-2 proteolytic activation by PIs at concentrations present in plasma of treated individuals. As PIs also inhibit the in vivo growth and invasion of an angiogenic tumor-cell line, these data indicate that PIs are potent anti-angiogenic and anti-tumor molecules that might be used in treating non-HIV KS and in other HIV-associated tumors. PMID- 11875493 TI - Non-hematopoietic allograft cells directly activate CD8+ T cells and trigger acute rejection: an alternative mechanism of allorecognition. AB - Despite evidence that human non-hematopoietic cells, such as vascular endothelium, can activate allogeneic T lymphocytes in vitro, the prevailing view has been that hematopoietic antigen-presenting cells are required to trigger alloimmune responses in vivo. Here we report that mouse non-hematopoietic cells activate alloreactive CD8+ T lymphocytes in vitro and in vivo. We also show that vascularized cardiac allografts are acutely rejected via CD8+ direct allorecognition even if the alloantigen is not presented by hematopoietic professional antigen-presenting cells. Because activation of alloreactive CD8+ T cells by donor-type non-hematopoietic cells can continue for the life of the allograft, these findings present a new clinically relevant mechanism of allorecognition and should be taken into consideration when developing strategies to prevent allograft vasculopathy or to induce tolerance. PMID- 11875494 TI - Heme oxygenase-1 mediates the anti-inflammatory effect of interleukin-10 in mice. AB - The mechanisms underlying the action of the potent anti-inflammatory interleukin 10 (IL-10) are poorly understood. Here we show that, in murine macrophages, IL-10 induces expression of heme oxygenase-1 (HO-1), a stress-inducible protein with potential anti-inflammatory effect, via a p38 mitogen-activated protein kinase dependent pathway. Inhibition of HO-1 protein synthesis or activity significantly reversed the inhibitory effect of IL-10 on production of tumor necrosis factor alpha induced by lipopolysaccharide (LPS). Additional experiments revealed the involvement of carbon monoxide, one of the products of HO-1-mediated heme degradation, in the anti-inflammatory effect of IL-10 in vitro. Induction of HO-1 by IL-10 was also evident in vivo. IL-10-mediated protection against LPS-induced septic shock in mice was significantly attenuated by cotreatment with the HO inhibitor, zinc protoporphyrin. The identification of HO-1 as a downstream effector of IL-10 provides new possibilities for improved therapeutic approaches for treating inflammatory diseases. PMID- 11875495 TI - CD40L stabilizes arterial thrombi by a beta3 integrin--dependent mechanism. AB - CD40L, a member of the tumor necrosis factor family of ligands, plays a major role in immune responses via its receptor, CD40. Recently, CD40L has been detected on the surfaces of activated platelets and shown to activate endothelium. Here we further addressed the function of platelet CD40L. We show that absence of CD40L affects the stability of arterial thrombi and delays arterial occlusion in vivo. Infusion of recombinant soluble (rs)CD40L restored normal thrombosis, whereas rsCD40L lacking the KGD integrin-recognition sequence did not. CD40-deficient mice exhibited normal thrombogenesis. rsCD40L specifically bound to purified integrin alphaIIbbeta3 and to activated platelets in a beta3-dependent manner and induced platelet spreading. In addition, rsCD40L promoted the aggregation of either human or mouse platelets under high shear rates. Thus, CD40L appears to be an alphaIIbbeta3 ligand, a platelet agonist, and necessary for stability of arterial thrombi. PMID- 11875496 TI - Modular flexibility of dystrophin: implications for gene therapy of Duchenne muscular dystrophy. AB - Attempts to develop gene therapy for Duchenne muscular dystrophy (DMD) have been complicated by the enormous size of the dystrophin gene. We have performed a detailed functional analysis of dystrophin structural domains and show that multiple regions of the protein can be deleted in various combinations to generate highly functional mini- and micro-dystrophins. Studies in transgenic mdx mice, a model for DMD, reveal that a wide variety of functional characteristics of dystrophy are prevented by some of these truncated dystrophins. Muscles expressing the smallest dystrophins are fully protected against damage caused by muscle activity and are not morphologically different from normal muscle. Moreover, injection of adeno-associated viruses carrying micro-dystrophins into dystrophic muscles of immunocompetent mdx mice results in a striking reversal of histopathological features of this disease. These results demonstrate that the dystrophic pathology can be both prevented and reversed by gene therapy using micro-dystrophins. PMID- 11875497 TI - Cloned mice have an obese phenotype not transmitted to their offspring. AB - Mammalian cloning using somatic cells has been accomplished successfully in several species, and its potential basic, clinical and therapeutic applications are being pursued on many fronts. Determining the long-term effects of cloning on offspring is crucial for consideration of future application of the technique. Although full-term development of animals cloned from adult somatic cells has been reported, problems in the resulting progeny indicate that the cloning procedure may not produce animals that are phenotypically identical to their cell donor. We used a mouse model to take advantage of its short generation time and lifespan. Here we report that the increased body weight of cloned B6C3F1 female mice reflects an increase of body fat in addition to a larger body size, and that these mice share many characteristics consistent with obesity. We also show that the obese phenotype is not transmitted to offspring generated by mating male and female cloned mice. PMID- 11875498 TI - Hematopoietic competence is a rare property of neural stem cells that may depend on genetic and epigenetic alterations. AB - The concept of stem-cell plasticity received strong support from a recent observation that extensively passaged, clonally derived neural stem cells could contribute to hematopoiesis. We investigated whether hematopoietic potential was a consistent or unusual feature of neural stem cells, and whether it depended on the extent of in vitro passaging before transplantation. Here we transplanted over 128 x 10(6) neurosphere cells into 128 host animals; however, we never observed contribution to hematopoiesis, irrespective of the number of passages and despite the use of an assay that could detect the contribution of a single blood stem cell to hematopoietic repopulation. Although extensively cultured neurosphere cells continued to generate neural progeny, marked changes in their growth properties occurred, including changes in growth-factor dependence, cell cycle kinetics, cell adhesion and gene expression. Our results exclude hematopoietic competence as a consistent property of intravenously infused neural stem cells. However, the consistent changes that occurred during extended passaging are compatible with genetic or epigenetic alterations and suggest that rare transformation events may account for the neural-to-blood fate switch originally reported. PMID- 11875499 TI - Tumor-cell resistance to death receptor--induced apoptosis through mutational inactivation of the proapoptotic Bcl-2 homolog Bax. AB - The importance of Bax for induction of tumor apoptosis through death receptors remains unclear. Here we show that Bax can be essential for death receptor- mediated apoptosis in cancer cells. Bax-deficient human colon carcinoma cells were resistant to death-receptor ligands, whereas Bax-expressing sister clones were sensitive. Bax was dispensable for apical death-receptor signaling events including caspase-8 activation, but crucial for mitochondrial changes and downstream caspase activation. Treatment of colon tumor cells deficient in DNA mismatch repair with the death-receptor ligand apo2 ligand (Apo2L)/tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) selected in vitro or in vivo for refractory subclones with Bax frameshift mutations including deletions at a novel site. Chemotherapeutic agents upregulated expression of the Apo2L/TRAIL receptor DR5 and the Bax homolog Bak in Baxminus sign/minus sign cells, and restored Apo2L/TRAIL sensitivity in vitro and in vivo. Thus, Bax mutation in mismatch repair--deficient tumors can cause resistance to death receptor--targeted therapy, but pre-exposure to chemotherapy rescues tumor sensitivity. PMID- 11875500 TI - Restoration of the tumor suppressor function to mutant p53 by a low-molecular weight compound. AB - The tumor suppressor p53 inhibits tumor growth primarily through its ability to induce apoptosis. Mutations in p53 occur in at least 50% of human tumors. We hypothesized that reactivation of mutant p53 in such tumors should trigger massive apoptosis and eliminate the tumor cells. To test this, we screened a library of low-molecular-weight compounds in order to identify compounds that can restore wild-type function to mutant p53. We found one compound capable of inducing apoptosis in human tumor cells through restoration of the transcriptional transactivation function to mutant p53. This molecule, named PRIMA-1, restored sequence-specific DNA binding and the active conformation to mutant p53 proteins in vitro and in living cells. PRIMA-1 rescued both DNA contact and structural p53 mutants. In vivo studies in mice revealed an antitumor effect with no apparent toxicity. This molecule may serve as a lead compound for the development of anticancer drugs targeting mutant p53. PMID- 11875501 TI - Prostaglandin E2 transactivates EGF receptor: a novel mechanism for promoting colon cancer growth and gastrointestinal hypertrophy. AB - Prostaglandins (PGs), bioactive lipid molecules produced by cyclooxygenase enzymes (COX-1 and COX-2), have diverse biological activities, including growth promoting actions on gastrointestinal mucosa. They are also implicated in the growth of colonic polyps and cancers. However, the precise mechanisms of these trophic actions of PGs remain unclear. As activation of the epidermal growth factor receptor (EGFR) triggers mitogenic signaling in gastrointestinal mucosa, and its expression is also upregulated in colonic cancers and most neoplasms, we investigated whether PGs transactivate EGFR. Here we provide evidence that prostaglandin E2 (PGE2) rapidly phosphorylates EGFR and triggers the extracellular signal-regulated kinase 2 (ERK2)--mitogenic signaling pathway in normal gastric epithelial (RGM1) and colon cancer (Caco-2, LoVo and HT-29) cell lines. Inactivation of EGFR kinase with selective inhibitors significantly reduces PGE2-induced ERK2 activation, c-fos mRNA expression and cell proliferation. Inhibition of matrix metalloproteinases (MMPs), transforming growth factor-alpha (TGF-alpha) or c-Src blocked PGE2-mediated EGFR transactivation and downstream signaling indicating that PGE2-induced EGFR transactivation involves signaling transduced via TGF-alpha, an EGFR ligand, likely released by c-Src-activated MMP(s). Our findings that PGE2 transactivates EGFR reveal a previously unknown mechanism by which PGE2 mediates trophic actions resulting in gastric and intestinal hypertrophy as well as growth of colonic polyps and cancers. PMID- 11875508 TI - Proteomics in a small world. PMID- 11875502 TI - Autoantigen microarrays for multiplex characterization of autoantibody responses. AB - We constructed miniaturized autoantigen arrays to perform large-scale multiplex characterization of autoantibody responses directed against structurally diverse autoantigens, using submicroliter quantities of clinical samples. Autoantigen microarrays were produced by attaching hundreds of proteins, peptides and other biomolecules to the surface of derivatized glass slides using a robotic arrayer. Arrays were incubated with patient serum, and spectrally resolvable fluorescent labels were used to detect autoantibody binding to specific autoantigens on the array. We describe and characterize arrays containing the major autoantigens in eight distinct human autoimmune diseases, including systemic lupus erythematosus and rheumatoid arthritis. This represents the first report of application of such technology to multiple human disease sera, and will enable validated detection of antibodies recognizing autoantigens including proteins, peptides, enzyme complexes, ribonucleoprotein complexes, DNA and post-translationally modified antigens. Autoantigen microarrays represent a powerful tool to study the specificity and pathogenesis of autoantibody responses, and to identify and define relevant autoantigens in human autoimmune diseases. PMID- 11875509 TI - An intermediate seeks instant gratification. PMID- 11875510 TI - Man bites dog. PMID- 11875511 TI - How a rotavirus hijacks the human protein synthesis machinery. PMID- 11875512 TI - Changing chromatin from the inside. PMID- 11875513 TI - Picture story. A DNA look-alike. PMID- 11875514 TI - Don C. Wiley. PMID- 11875515 TI - Crystal structures of ferredoxin variants exhibiting large changes in [Fe-S] reduction potential. AB - Elucidating how proteins control the reduction potentials (E0') of [Fe--S] clusters is a longstanding fundamental problem in bioinorganic chemistry. Two site-directed variants of Azotobacter vinelandii ferredoxin I (FdI) that show large shifts in [Fe--S] cluster E0' (100--200 mV versus standard hydrogen electrode (SHE)) have been characterized. High resolution X-ray structures of F2H and F25H variants in their oxidized forms, and circular dichroism (CD) and electron paramagnetic resonance (EPR) of the reduced forms indicate that the overall structure is not affected by the mutations and reveal that there is no increase in solvent accessibility nor any reorientation of backbone amide dipoles or NH--S bonds. The structures, combined with detailed investigation of the variation of E0' with pH and temperature, show that the largest increases in E0' result from the introduction of positive charge due to protonation of the introduced His residues. The smaller (50--100 mV) increases observed for the neutral form are proposed to occur by directing a Hdelta+--Ndelta- dipole toward the reduced form of the cluster. PMID- 11875516 TI - Im7 folding mechanism: misfolding on a path to the native state. AB - Many proteins populate collapsed intermediate states during folding. In order to elucidate the nature and importance of these species, we have mapped the structure of the on-pathway intermediate of the four-helix protein, Im7, together with the conformational changes it undergoes as it folds to the native state. Kinetic data for 29 Im7 point mutants show that the intermediate contains three of the four helices found in the native structure, packed around a specific hydrophobic core. However, the intermediate contains many non-native interactions; as a result, hydrophobic interactions become disrupted in the rate limiting transition state before the final helix docks onto the developing structure. The results of this study support a hierarchical mechanism of protein folding and explain why the misfolding of Im7 occurs. The data also demonstrate that non-native interactions can play a significant role in folding, even for small proteins with simple topologies. PMID- 11875518 TI - Solution structure of the DNA-binding domain of MafG. AB - The Maf family proteins, which constitute a subgroup of basic region-leucine zipper (bZIP) proteins, function as transcriptional regulators of cellular differentiation. Together with the basic region, the Maf extended homology region (EHR), conserved only within the Maf family, defines the DNA binding specific to Mafs. Here we present the first NMR-derived structure of the DNA-binding domain (residues 1-76) of MafG, which contains the EHR and the basic region. The structure consists of three alpha-helices and resembles the fold of the DNA binding domain of Skn-1, a developmental transcription factor of Caenorhabditis elegans. The structural similarity between MafG and Skn-1 enables us to propose a possible mechanism by which Maf family proteins recognize their consensus DNA sequences. PMID- 11875519 TI - Structure of the C-terminal FG-nucleoporin binding domain of Tap/NXF1. AB - The vertebrate Tap protein is a member of the NXF family of shuttling transport receptors for nuclear export of mRNA. Tap has a modular structure, and its most C terminal domain is important for binding to FG repeat-containing nuclear pore proteins (FG-nucleoporins) and is sufficient to mediate nuclear shuttling. We report the solution structure of this C-terminal domain, which is based on a distinctive arrangement of four alpha-helices and is joined to the next module by a flexible 12-residue Pro-rich linker. F617A Tap suppresses FG-nucleoporin binding by the most C-terminal domain that, together with the structure of the other modules from which Tap is constructed, provides a structural context for its nuclear shuttling function. PMID- 11875520 TI - The crystal structure of class II ribonucleotide reductase reveals how an allosterically regulated monomer mimics a dimer. AB - Ribonucleotide reductases (RNRs) catalyze the conversion of ribonucleotides to deoxyribonucleotides, an essential step in DNA biosynthesis and repair. Here we present the crystal structure of class II (coenzyme B12-dependent) ribonucleoside triphosphate reductase (RTPR) from Lactobacillus leichmannii in the apo enzyme form and in complex with the B12 analog adeninylpentylcobalamin at 1.75 and 2.0 A resolution, respectively. This monomeric, allosterically regulated class II RNR retains all the key structural features associated with the catalytic and regulatory machinery of oligomeric RNRs. Surprisingly, the dimer interface responsible for effector binding in class I RNR is preserved through a single 130 residue insertion in the class II structure. Thus, L. leichmannii RNR is a paradigm for the simplest structural entity capable of ribonucleotide reduction, a reaction linking the RNA and DNA worlds. PMID- 11875522 TI - At last, a chance for postdocs to learn how to teach. PMID- 11875524 TI - A blast from the past. PMID- 11875525 TI - Burdened by expectation. PMID- 11875526 TI - Nuclear-weapons design plan raises fresh proliferation fears. PMID- 11875527 TI - Foreign researchers turn their backs on Germany. PMID- 11875528 TI - Minimum standards set out for gene-expression data. PMID- 11875529 TI - Power vacuum expands as CDC director resigns. PMID- 11875530 TI - Reef under threat from 'bleaching' outbreak. PMID- 11875531 TI - Academy proposes tighter crop monitoring. PMID- 11875532 TI - Alleged flaws in gene-transfer paper spark row over genetically modified maize. PMID- 11875533 TI - Poor nations seek new biodiversity deal. PMID- 11875534 TI - Cutbacks cost jobs at agricultural institute. PMID- 11875536 TI - Rebirth and regeneration. PMID- 11875537 TI - Voyage of the argonauts. PMID- 11875538 TI - Locking horns. PMID- 11875539 TI - Physics gets physical. PMID- 11875540 TI - Why impact factors don't work for taxonomy. PMID- 11875544 TI - The future of sex. PMID- 11875545 TI - Magnetic moments at Jupiter. PMID- 11875548 TI - The mass question. PMID- 11875546 TI - A monoclonal mouse? PMID- 11875549 TI - Walking with tyrannosaurs. PMID- 11875550 TI - Slip-sliding away. PMID- 11875552 TI - Mellifluous matures to malodorous in musth. AB - Male Asian elephants in musth--an annual period of heightened sexual activity and intensified aggression--broadcast odoriferous, behaviourally influential messages from secretions of the temporal gland. From our observations in the wild, together with instantaneous chemical sampling and captive-elephant playback experiments, we have discovered that young, socially immature males in musth signal their naivety by releasing honey-like odours to avoid conflict with adult males, whereas older musth males broadcast malodorous combinations to deter young males, facilitating the smooth functioning of male society. As elephant--human conflicts can upset this equilibrium, chemically modulating male behaviour may be one way to help the conservation of wild elephants. PMID- 11875553 TI - Age of long sediment cores from Lake Baikal. AB - The new BDP-98 drill core of the Baikal Drilling Project is a key palaeoclimate record in continental Asia because globally sensitive sedimentary records of such length and continuity are very rare. Kashiwaya et al. have attempted signal processing of the BDP-98 average grain-size record, but in constructing their age model they excised a 100-metre interval from the 600-metre section, stating that it is "erroneous". On the basis of our lithological studies, we consider that this excision is unjustified. PMID- 11875555 TI - MAP kinase signalling cascade in Arabidopsis innate immunity. AB - There is remarkable conservation in the recognition of pathogen-associated molecular patterns (PAMPs) by innate immune responses of plants, insects and mammals. We developed an Arabidopsis thaliana leaf cell system based on the induction of early-defence gene transcription by flagellin, a highly conserved component of bacterial flagella that functions as a PAMP in plants and mammals. Here we identify a complete plant MAP kinase cascade (MEKK1, MKK4/MKK5 and MPK3/MPK6) and WRKY22/WRKY29 transcription factors that function downstream of the flagellin receptor FLS2, a leucine-rich-repeat (LRR) receptor kinase. Activation of this MAPK cascade confers resistance to both bacterial and fungal pathogens, suggesting that signalling events initiated by diverse pathogens converge into a conserved MAPK cascade. PMID- 11875556 TI - Control of Jupiter's radio emission and aurorae by the solar wind. AB - Radio emissions from Jupiter provided the first evidence that this giant planet has a strong magnetic field and a large magnetosphere. Jupiter also has polar aurorae, which are similar in many respects to Earth's aurorae. The radio emissions are believed to be generated along the high-latitude magnetic field lines by the same electrons that produce the aurorae, and both the radio emission in the hectometric frequency range and the aurorae vary considerably. The origin of the variability, however, has been poorly understood. Here we report simultaneous observations using the Cassini and Galileo spacecraft of hectometric radio emissions and extreme ultraviolet auroral emissions from Jupiter. Our results show that both of these emissions are triggered by interplanetary shocks propagating outward from the Sun. When such a shock arrives at Jupiter, it seems to cause a major compression and reconfiguration of the magnetosphere, which produces strong electric fields and therefore electron acceleration along the auroral field lines, similar to the processes that occur during geomagnetic storms at the Earth. PMID- 11875557 TI - Ultra-relativistic electrons in Jupiter's radiation belts. AB - Ground-based observations have shown that Jupiter is a two-component source of microwave radio emission: thermal atmospheric emission and synchrotron emission from energetic electrons spiralling in Jupiter's magnetic field. Later in situ measurements confirmed the existence of Jupiter's high-energy electron-radiation belts, with evidence for electrons at energies up to 20[?]MeV. Although most radiation belt models predict electrons at higher energies, adiabatic diffusion theory can account only for energies up to around 20[?]MeV. Unambiguous evidence for more energetic electrons is lacking. Here we report observations of 13.8[?]GHz synchrotron emission that confirm the presence of electrons with energies up to 50[?]MeV; the data were collected during the Cassini fly-by of Jupiter. These energetic electrons may be repeatedly accelerated through an interaction with plasma waves, which can transfer energy into the electrons. Preliminary comparison of our data with model results suggests that electrons with energies of less than 20[?]MeV are more numerous than previously believed. PMID- 11875558 TI - The dusk flank of Jupiter's magnetosphere. AB - Limited single-spacecraft observations of Jupiter's magnetopause have been used to infer that the boundary moves inward or outward in response to variations in the dynamic pressure of the solar wind. At Earth, multiple-spacecraft observations have been implemented to understand the physics of how this motion occurs, because they can provide a snapshot of a transient event in progress. Here we present a set of nearly simultaneous two-point measurements of the jovian magnetopause at a time when the jovian magnetopause was in a state of transition from a relatively larger to a relatively smaller size in response to an increase in solar-wind pressure. The response of Jupiter's magnetopause is very similar to that of the Earth, confirming that the understanding built on studies of the Earth's magnetosphere is valid. The data also reveal evidence for a well developed boundary layer just inside the magnetopause. PMID- 11875559 TI - A nebula of gases from Io surrounding Jupiter. AB - Several planetary missions have reported the presence of substantial numbers of energetic ions and electrons surrounding Jupiter; relativistic electrons are observable up to several astronomical units (au) from the planet. A population of energetic (>30[?]keV) neutral particles also has been reported, but the instrumentation was not able to determine the mass or charge state of the particles, which were subsequently labelled energetic neutral atoms. Although images showing the presence of the trace element sodium were obtained, the source and identity of the neutral atoms---and their overall significance relative to the loss of charged particles from Jupiter's magnetosphere---were unknown. Here we report the discovery by the Cassini spacecraft of a fast (>103[?]km[?]s-1) and hot magnetospheric neutral wind extending more than 0.5[?]au from Jupiter, and the presence of energetic neutral atoms (both hot and cold) that have been accelerated by the electric field in the solar wind. We suggest that these atoms originate in volcanic gases from Io, undergo significant evolution through various electromagnetic interactions, escape Jupiter's magnetosphere and then populate the environment around the planet. Thus a 'nebula' is created that extends outwards over hundreds of jovian radii. PMID- 11875560 TI - Ultraviolet emissions from the magnetic footprints of Io, Ganymede and Europa on Jupiter. AB - Io leaves a magnetic footprint on Jupiter's upper atmosphere that appears as a spot of ultraviolet emission that remains fixed underneath Io as Jupiter rotates. The specific physical mechanisms responsible for generating those emissions are not well understood, but in general the spot seems to arise because of an electromagnetic interaction between Jupiter's magnetic field and the plasma surrounding Io, driving currents of around 1 million amperes down through Jupiter's ionosphere. The other galilean satellites may also leave footprints, and the presence or absence of such footprints should illuminate the underlying physical mechanism by revealing the strengths of the currents linking the satellites to Jupiter. Here we report persistent, faint, far-ultraviolet emission from the jovian footprints of Ganymede and Europa. We also show that Io's magnetic footprint extends well beyond the immediate vicinity of Io's flux-tube interaction with Jupiter, and much farther than predicted theoretically; the emission persists for several hours downstream. We infer from these data that Ganymede and Europa have persistent interactions with Jupiter's magnetic field despite their thin atmospheres. PMID- 11875561 TI - A pulsating auroral X-ray hot spot on Jupiter. AB - Jupiter's X-ray aurora has been thought to be excited by energetic sulphur and oxygen ions precipitating from the inner magnetosphere into the planet's polar regions. Here we report high-spatial-resolution observations that demonstrate that most of Jupiter's northern auroral X-rays come from a 'hot spot' located significantly poleward of the latitudes connected to the inner magnetosphere. The hot spot seems to be fixed in magnetic latitude and longitude and occurs in a region where anomalous infrared and ultraviolet emissions have also been observed. We infer from the data that the particles that excite the aurora originate in the outer magnetosphere. The hot spot X-rays pulsate with an approximately 45-min period, a period similar to that reported for high-latitude radio and energetic electron bursts observed by near-Jupiter spacecraft. These results invalidate the idea that jovian auroral X-ray emissions are mainly excited by steady precipitation of energetic heavy ions from the inner magnetosphere. Instead, the X-rays seem to result from currently unexplained processes in the outer magnetosphere that produce highly localized and highly variable emissions over an extremely wide range of wavelengths. PMID- 11875562 TI - Transient aurora on Jupiter from injections of magnetospheric electrons. AB - Energetic electrons and ions that are trapped in Earth's magnetosphere can suddenly be accelerated towards the planet. Some dynamic features of Earth's aurora (the northern and southern lights) are created by the fraction of these injected particles that travels along magnetic field lines and hits the upper atmosphere. Jupiter's aurora appears similar to Earth's in some respects; both appear as large ovals circling the poles and both show transient events. But the magnetospheres of Jupiter and Earth are so different---particularly in the way they are powered---that it is not known whether the magnetospheric drivers of Earth's aurora also cause them on Jupiter. Here we show a direct relationship between Earth-like injections of electrons in Jupiter's magnetosphere and a transient auroral feature in Jupiter's polar region. This relationship is remarkably similar to what happens at Earth, and therefore suggests that despite the large differences between planetary magnetospheres, some processes that generate aurorae are the same throughout the Solar System. PMID- 11875563 TI - Bandgap modulation of carbon nanotubes by encapsulated metallofullerenes. AB - Motivated by the technical and economic difficulties in further miniaturizing silicon-based transistors with the present fabrication technologies, there is a strong effort to develop alternative electronic devices, based, for example, on single molecules. Recently, carbon nanotubes have been successfully used for nanometre-sized devices such as diodes, transistors, and random access memory cells. Such nanotube devices are usually very long compared to silicon-based transistors. Here we report a method for dividing a semiconductor nanotube into multiple quantum dots with lengths of about 10nm by inserting Gd@C82 endohedral fullerenes. The spatial modulation of the nanotube electronic bandgap is observed with a low-temperature scanning tunnelling microscope. We find that a bandgap of approximately 0.5eV is narrowed down to approximately 0.1eV at sites where endohedral metallofullerenes are inserted. This change in bandgap can be explained by local elastic strain and charge transfer at metallofullerene sites. This technique for fabricating an array of quantum dots could be used for nano electronics and nano-optoelectronics. PMID- 11875564 TI - Factors determining crystal--liquid coexistence under shear. AB - The interaction between an imposed shear flow and an order--disorder transition underlies a broad range of phenomena. Under the influence of shear flow, a variety of soft matter is observed to spontaneously form bands characterized by different local order---for example, thermotropic liquid crystals subjected to shear flow exhibit rich phase behaviour. The stability of order under the influence of shear flow is also fundamental to understanding frictional wear and lubrication. Although there exists a well developed theoretical approach to the influence of shear flow on continuous transitions in fluid mixtures, little is known about the underlying principles governing non-equilibrium coexistence between phases of different symmetry. Here we show, using non-equilibrium molecular dynamics simulations of a system of spherical particles, that a stationary coexistence exists between a strained crystal and the shearing liquid, and that this coexistence cannot be accounted for by invoking a non-equilibrium analogue of the chemical potential. Instead of such thermodynamic arguments, we argue that a balancing of the crystal growth rate with the rate of surface erosion by the shearing melt can account for the observed coexistence. PMID- 11875565 TI - High mixing rates in the abyssal Southern Ocean. AB - Mixing of water masses from the deep ocean to the layers above can be estimated from considerations of continuity in the global ocean overturning circulation. But averaged over ocean basins, diffusivity has been observed to be too small to account for the global upward flux of water, and high mixing intensities have only been found in the restricted areas close to sills and narrow gaps. Here we present observations from the Scotia Sea, a deep ocean basin between the Antarctic peninsula and the tip of South America, showing a high intensity of mixing that is unprecedented over such a large area. Using a budget calculation over the whole basin, we find a diffusivity of (39 plus minus 10) x 104[?]m2[?]s 1, averaged over an area of 7 x 105[?]km2. The Scotia Sea is a basin with a rough topography, situated just east of the Drake passage where the strong flow of the Antarctic Circumpolar Current is constricted in width. The high basin-wide mixing intensity in this area of the Southern Ocean may help resolve the question of where the abyssal water masses are mixed towards the surface. PMID- 11875566 TI - Sudden aseismic fault slip on the south flank of Kilauea volcano. AB - One of the greatest hazards associated with oceanic volcanoes is not volcanic in nature, but lies with the potential for catastrophic flank failure. Such flank failure can result in devastating tsunamis and threaten not only the immediate vicinity, but coastal cities along the entire rim of an ocean basin. Kilauea volcano on the island of Hawaii, USA, is a potential source of such flank failures and has therefore been monitored by a network of continuously recording geodetic instruments, including global positioning system (GPS) receivers, tilt meters and strain meters. Here we report that, in early November 2000, this network recorded transient southeastward displacements, which we interpret as an episode of aseismic fault slip. The duration of the event was about 36 hours, it had an equivalent moment magnitude of 5.7 and a maximum slip velocity of about 6[?]cm per day. Inversion of the GPS data reveals a shallow-dipping thrust fault at a depth of 4.5[?]km that we interpret as the down-dip extension of the Hilina Pali--Holei Pali normal fault system. This demonstrates that continuously recording geodetic networks can detect accelerating slip, potentially leading to warnings of volcanic flank collapse. PMID- 11875567 TI - Tyrannosaurus was not a fast runner. AB - The fastest gait and speed of the largest theropod (carnivorous) dinosaurs, such as Tyrannosaurus, is controversial. Some studies contend that Tyrannosaurus was limited to walking, or at best an 11 m s(-1) top speed, whereas others argue for at least 20 m s(-1) running speeds. We demonstrate a method of gauging running ability by estimating the minimum mass of extensor (supportive) muscle needed for fast running. The model's predictions are validated for living alligators and chickens. Applying the method to small dinosaurs corroborates other studies by showing that they could have been competent runners. However, models show that in order to run quickly, an adult Tyrannosaurus would have needed an unreasonably large mass of extensor muscle, even with generous assumptions. Therefore, it is doubtful that Tyrannosaurus and other huge dinosaurs (approximately 6,000 kg) were capable runners or could reach high speeds. PMID- 11875568 TI - Adaptive protein evolution in Drosophila. AB - For over 30 years a central question in molecular evolution has been whether natural selection plays a substantial role in evolution at the DNA sequence level. Evidence has accumulated over the last decade that adaptive evolution does occur at the protein level, but it has remained unclear how prevalent adaptive evolution is. Here we present a simple method by which the number of adaptive substitutions can be estimated and apply it to data from Drosophila simulans and D. yakuba. We estimate that 45% of all amino-acid substitutions have been fixed by natural selection, and that on average one adaptive substitution occurs every 45 years in these species. PMID- 11875569 TI - Testing the neutral theory of molecular evolution with genomic data from Drosophila. AB - Although positive selection has been detected in many genes, its overall contribution to protein evolution is debatable. If the bulk of molecular evolution is neutral, then the ratio of amino-acid (A) to synonymous (S) polymorphism should, on average, equal that of divergence. A comparison of the A/S ratio of polymorphism in Drosophila melanogaster with that of divergence from Drosophila simulans shows that the A/S ratio of divergence is twice as high---a difference that is often attributed to positive selection. But an increase in selective constraint owing to an increase in effective population size could also explain this observation, and, if so, all genes should be affected similarly. Here we show that the difference between polymorphism and divergence is limited to only a fraction of the genes, which are also evolving more rapidly, and this implies that positive selection is responsible. A higher A/S ratio of divergence than of polymorphism is also observed in other species, which suggests a rate of adaptive evolution that is far higher than permitted by the neutral theory of molecular evolution. PMID- 11875570 TI - Brain potential and functional MRI evidence for how to handle two languages with one brain. AB - Bilingual individuals need effective mechanisms to prevent interference from one language while processing material in the other. Here we show, using event related brain potentials and functional magnetic resonance imaging (fMRI), that words from the non-target language are rejected at an early stage before semantic analysis in bilinguals. Bilingual Spanish/Catalan and monolingual Spanish subjects were instructed to press a button when presented with words in one language, while ignoring words in the other language and pseudowords. The brain potentials of bilingual subjects in response to words of the non-target language were not sensitive to word frequency, indicating that the meaning of non-target words was not accessed in bilinguals. The fMRI activation patterns of bilinguals included a number of areas previously implicated in phonological and pseudoword processing, suggesting that bilinguals use an indirect phonological access route to the lexicon of the target language to avoid interference. PMID- 11875571 TI - Functional neurogenesis in the adult hippocampus. AB - There is extensive evidence indicating that new neurons are generated in the dentate gyrus of the adult mammalian hippocampus, a region of the brain that is important for learning and memory. However, it is not known whether these new neurons become functional, as the methods used to study adult neurogenesis are limited to fixed tissue. We use here a retroviral vector expressing green fluorescent protein that only labels dividing cells, and that can be visualized in live hippocampal slices. We report that newly generated cells in the adult mouse hippocampus have neuronal morphology and can display passive membrane properties, action potentials and functional synaptic inputs similar to those found in mature dentate granule cells. Our findings demonstrate that newly generated cells mature into functional neurons in the adult mammalian brain. PMID- 11875572 TI - Monoclonal mice generated by nuclear transfer from mature B and T donor cells. AB - Cloning from somatic cells is inefficient, with most clones dying during gestation. Cloning from embryonic stem (ES) cells is much more effective, suggesting that the nucleus of an embryonic cell is easier to reprogram. It is thus possible that most surviving clones are, in fact, derived from the nuclei of rare somatic stem cells present in adult tissues, rather than from the nuclei of differentiated cells, as has been assumed. Here we report the generation of monoclonal mice by nuclear transfer from mature lymphocytes. In a modified two step cloning procedure, we established ES cells from cloned blastocysts and injected them into tetraploid blastocysts to generate mice. In this approach, the embryo is derived from the ES cells and the extra-embryonic tissues from the tetraploid host. Animals cloned from a B-cell nucleus were viable and carried fully rearranged immunoglobulin alleles in all tissues. Similarly, a mouse cloned from a T-cell nucleus carried rearranged T-cell-receptor genes in all tissues. This is an unequivocal demonstration that a terminally differentiated cell can be reprogrammed to produce an adult cloned animal. PMID- 11875573 TI - MEC-2 regulates C. elegans DEG/ENaC channels needed for mechanosensation. AB - Touch sensitivity in animals relies on nerve endings in the skin that convert mechanical force into electrical signals. In the nematode Caenorhabditis elegans, gentle touch to the body wall is sensed by six mechanosensory neurons that express two amiloride-sensitive Na+ channel proteins (DEG/ENaC). These proteins, MEC-4 and MEC-10, are required for touch sensation and can mutate to cause neuronal degeneration. Here we show that these mutant or 'd' forms of MEC-4 and MEC-10 produce a constitutively active, amiloride-sensitive ionic current when co expressed in Xenopus oocytes, but not on their own. MEC-2, a stomatin-related protein needed for touch sensitivity, increased the activity of mutant channels about 40-fold and allowed currents to be detected with wild-type MEC-4 and MEC 10. Whereas neither the central, stomatin-like domain of MEC-2 nor human stomatin retained the activity of full-length MEC-2, both produced amiloride-sensitive currents with MEC-4d. Our findings indicate that MEC-2 regulates MEC-4/MEC-10 ion channels and raise the possibility that similar ion channels may be formed by stomatin-like proteins and DEG/ENaC proteins that are co-expressed in both vertebrates and invertebrates. Some of these channels may mediate mechanosensory responses. PMID- 11875574 TI - Characterization of a common precursor population for dendritic cells. AB - Dendritic cells (DCs) are essential for the establishment of immune responses against pathogens and tumour cells, and thus have great potential as tools for vaccination and cancer immunotherapy trials. Experimental evidence has led to a dual DC differentiation model, which involves the existence of both myeloid- and lymphoid-derived DCs. But this concept has been challenged by recent reports demonstrating that both CD8- and CD8+ DCs, considered in mice as archetypes of myeloid and lymphoid DCs respectively, can be generated from either lymphoid or myeloid progenitors. The issue of DC physiological derivation therefore remains an open question. Here we report the characterization of a DC-committed precursor population, which has the capacity to generate all the DC subpopulations present in mouse lymphoid organs---including CD8- and CD8+ DCs, as well as the B220+ DC subset---but which is devoid of myeloid or lymphoid differentiation potential. These data support an alternative model of DC development, in which there is an independent, common DC differentiation pathway. PMID- 11875575 TI - A blue-light-activated adenylyl cyclase mediates photoavoidance in Euglena gracilis. AB - Blue light regulates processes such as the development of plants and fungi and the behaviour of microbes. Two types of blue-light receptor flavoprotein have been identified: cryptochromes, which have partial similarity to photolyases, and phototropins, which are photoregulated protein kinases. The former have also been found in animals with evidence of essential roles in circadian rhythms. Euglena gracilis, a unicellular flagellate, abruptly changes its swimming direction after a sudden increase or decrease in incident blue light intensity, that is, step-up or step-down photophobic responses, resulting in photoavoidance or photoaccumulation, respectively. Although these photobehaviours of Euglena have been studied for a century, the photoreceptor molecules mediating them have remained unknown. Here we report the discovery and biochemical characterization of a new type of blue-light receptor flavoprotein, photoactivated adenylyl cyclase, in the photoreceptor organelle of Euglena gracilis, with molecular genetic evidence that it mediates the step-up photophobic response. PMID- 11875576 TI - Structure and dynamics of KH domains from FBP bound to single-stranded DNA. AB - Gene regulation can be tightly controlled by recognition of DNA deformations that are induced by stress generated during transcription. The KH domains of the FUSE binding protein (FBP), a regulator of c-myc expression, bind in vivo and in vitro to the single-stranded far-upstream element (FUSE), 1,500 base pairs upstream from the c-myc promoter. FBP bound to FUSE acts through TFIIH at the promoter. Here we report the solution structure of a complex between the KH3 and KH4 domains of FBP and a 29-base single-stranded DNA from FUSE. The KH domains recognize two sites, 9-10 bases in length, separated by 5 bases, with KH4 bound to the 5' site and KH3 to the 3' site. The central portion of each site comprises a tetrad of sequence 5'd-ATTC for KH4 and 5'd-TTTT for KH3. Dynamics measurements show that the two KH domains bind as articulated modules to single-stranded DNA, providing a flexible framework with which to recognize transient, moving targets. PMID- 11875579 TI - Ask not what you can do for us, but what we can do for you. PMID- 11875580 TI - Children and heart health: the 2002 annual report card on Canadians' health. PMID- 11875581 TI - Weathering the storm. PMID- 11875582 TI - Magnesium sulphate infusion suppresses the cardiac release of noradrenaline during a handgrip stress test. AB - BACKGROUND: Magnesium has several important cardiovascular effects, but its effect on cardiac sympathetic efferent neuron activity has not been clarified. OBJECTIVES: To examine the effect of magnesium sulphate infusion on cardiac sympathetic efferent postganglionic neuronal liberation of noradrenaline. PATIENTS AND METHODS: Twenty-two patients who underwent cardiac catheterization were randomly allocated to the control group or the magnesium group. Plasma noradrenaline and adrenaline concentrations in the aorta and the coronary sinus were measured. Noradrenaline or adrenaline release from the heart was calculated by dividing the difference in noradrenaline or adrenaline concentration between the aorta and the coronary sinus by that of the aorta. After baseline blood sampling, the control patients and the patients in the magnesium group received intravenous infusion of saline or magnesium sulphate (10 mmol). All patients were then subjected to 3 min of handgrip exercise stress test to augment sympathetic efferent neuronal activity, and the blood sampling was repeated. RESULTS: Although blood pressure was increased by the handgrip stress test, there were no differences in heart rate and blood pressure between the two groups, both at baseline and during the handgrip stress test. The plasma noradrenaline and adrenaline concentrations and noradrenaline or adrenaline release from the heart did not differ between the two groups in the baseline condition. However, the handgrip stress increased plasma noradrenaline concentrations and the cardiac noradrenaline release was increased in the control group, whereas the cardiac noradrenaline release was not increased by the handgrip stress in the magnesium group (P<0.02). CONCLUSIONS: These data indicate that magnesium sulphate infusion suppresses the release of catecholamines by the heart, which is an indirect index of sympathetic efferent neuronal activity. PMID- 11875584 TI - Effects of the 1994 Canadian Cardiovascular Society clinical practice guidelines for congestive heart failure. AB - BACKGROUND: In 1994, the Canadian Cardiovascular Society (CCS) issued practice guidelines for the management of congestive heart failure (CHF), which strongly recommended the use of angiotensin-converting enzyme (ACE) inhibitors. OBJECTIVE: To compare a strategy of active implementation of the CCS guidelines for CHF with the usual passive approach on the use of ACE inhibitors in hospitalized patients with CHF. PATIENTS AND METHODS: The study was conducted in eight Canadian hospitals close to the time of release of the CCS guidelines for CHF in the summer of 1994. The patients comprised those who were admitted to the participating hospitals with a diagnosis of CHF during the study period. Active dissemination included stakeholder development of pocket cards outlining an algorithm for the management of CHF based on the CCS guidelines and was conducted at the two Edmonton, Alberta hospitals. The cards were widely disseminated in conjunction with a series of workshops and presentations. Passive dissemination occurred at six other hospitals, and involved only the distribution of the CCS guidelines in the usual fashion (with no structured dissemination program). The primary outcome measured was the use of ACE inhibitors in the six- to 12-month period before the release of the CCS guidelines, compared with after the release in the active and passive groups. RESULTS: In the active group, hospital records of 1170 patients with CHF were reviewed before, and 1279 were reviewed after, the release of the CCS guidelines. In the passive group, 3436 were reviewed before, and 1912 were reviewed after the release of the guidelines. ACE inhibitor use did not change significantly in the active group (52.4% before versus 50.9% after) or in the passive group (53.4% before versus 56.5% after). CONCLUSIONS: Neither the active nor passive approaches to the dissemination of the CCS guidelines for CHF had any impact on the use of ACE inhibitors in hospitalized patients with CHF. Further efforts to package, deliver and evaluate guidelines are needed. PMID- 11875583 TI - Transition of care to adult congenital heart centres: what do patients know about their heart condition? AB - BACKGROUND: The population of adults with congenital heart disease is growing rapidly, and the transition of care from the pediatric to the adult setting is often not optimal. OBJECTIVES: To assess the level of knowledge that adults with congenital heart disease have of their condition. METHODS: All new patients referred to an adult congenital heart centre were asked to answer questions about their clinical diagnosis, their need and reason for anti-bioprophylaxis, and their risks of pregnancy when applicable. The patient's age at first visit, source of referral and time from last visit in cardiology (defined as the last visit with a cardiologist) were also noted. RESULTS: From November 1999 to July 2000, 104 patients (42 men) were referred to the adult centre by pediatric cardiologists (56), medical cardiologists (26), general practitioners (11), dentists (two) or obstetricians (two), or they were self-referred (seven). The mean age at the time of referral was 28 11 years (range 16 to 72 years, median 24 years). The time from the last visit in cardiology varied widely from one month to 25 years (median three years), with 29 patients (28%) having had no follow-up for more than five years. Among these, 14 patients had no follow-up for more than 10 years, with six patients having been referred for complications related to their cardiac anatomy, such as heart failure (Eisenmenger, Ebstein), syncope (operated and unoperated tetralogy of Fallot) and arrhythmias (atrial septal defect, unoperated Fallot). The clinical diagnosis was completely unknown by 36 patients (34.6%), including patients with repaired Fallot (three patients), Mustard procedure (two), severe aortic stenosis (two), severe pulmonary stenosis (one), Eisenmenger (one), unoperated Fallot (one), ventricular septal defect (six) and bicuspid aortic valve (seven). Seventy-three patients (79%) knew about antibiotic prophylaxis, but 50% did not comprehend why they needed it. Many women (66%) had never discussed the risks of pregnancy with their physician. CONCLUSIONS: A large percentage of adults with congenital heart abnormalities show a poor level of knowledge about their heart condition. These observations suggest the importance of structured transitional programs, the impact of which will need to be validated in prospective studies. PMID- 11875585 TI - Effects of chronic, rapid right atrial pacing on cardiac hemodynamics and myofibrillar ATPase activity in piglets. AB - OBJECTIVES: To determine whether chronic, rapid right atrial pacing in newborn neonatal piglets has any effects on cardiac hemodynamics, and whether these changes are associated with intrinsic alterations in cardiac contractile potential as shown by cardiac myofibrillar calcium ATPase activity. BACKGROUND: Although many studies have examined aspects of heart function in models of supraventricular tachycardia, far less is known about its effects in neonatal animals. It is thought that rapid pacing induces a dilated cardiomyopathy in immature pigs. ANIMALS AND METHODS: Two-week-old piglets underwent rapid right atrial pacing (250 beats/min) for 10 days, and their cardiac hemodynamic response was monitored. To obtain subcellular mechanistic information regarding systolic dysfunction, cardiac myofibrils were isolated and calcium adenosine triphosphatase activity was measured. RESULTS: Control piglets had a heart rate of 185 beats/min at the end of the experimental period. Pulmonary artery flow, pulmonary artery flow index and left ventricular end-diastolic diameter were unchanged as a function of rapid, chronic right atrial pacing. Aortic pressure decreased in the paced piglets. Left atrial pressure increased approximately threefold in the paced animals. Left ventricular end-systolic diameter was also significantly higher after pacing, but left ventricular end-diastolic diameter was unchanged. Left ventricular shortening fraction was depressed approximately 50%. Myofibrillar calcium adenosine triphosphatase activity was significantly depressed as a function of pacing. CONCLUSIONS: Neonatal piglets undergoing chronic supraventricular tachycardia exhibit systolic dysfunction in the absence of dilation. The depression in contractile protein calcium adenosine triphosphatase activity provides information at a subcellular level regarding the mechanism responsible for this cardiomyopathy. PMID- 11875586 TI - Comparative effects of nitroglycerin on intestinal vascular capacitance and conductance. AB - BACKGROUND: Nitroglycerin (NTG) dilates capacitance veins and resistance arterioles, but its relative effects on veins and arterioles are not known. OBJECTIVES: To compare NTG-induced changes in capacitance and conductance. ANIMALS AND METHODS: Aortic, left ventricular and portal venous (P(port)) pressures, portal flow and relative changes in intestinal blood volume (IBV) ((99m)technetium blood-pool scintigraphy) were measured in seven isoflurane anesthetized, splenectomized dogs. Changes in intestinal vascular capacitance and conductance (mean portal flow/[mean aortic pressure - mean P(port)]) were determined when NTG was continuously administered (0.8 to 150 microg/kg/min) into a jugular vein. Pressure-volume (ie, P(port)-IBV) curves were defined by impeding portal flow, and capacitance was defined as the IBV at P(port)=7.5 mmHg. RESULTS: At lower doses, NTG increased capacitance without increasing conductance, but conductance increased considerably with little further increase in capacitance at higher doses. Dose-response analysis revealed that the half-maximum capacitance effect was achieved at an NTG infusion rate of 3.5 microg/kg/min, whereas a rate of 35 microg/kg/min was required for the half-maximum conductance effect. CONCLUSIONS: At lower doses, NTG dilates capacitance vessels primarily, and that effect approaches its maximum before significant dilation of conductance vessels is manifest. However, at higher doses, the increase in conductance is substantial with little additional effect on capacitance. PMID- 11875587 TI - Vasopeptidase inhibition: a novel approach to cardiovascular therapy. AB - Omapatrilat was designed to inhibit simultaneously angiotensin-converting enzyme (ACE) and neutral endopeptidase (NEP). The ubiquitous involvement of the renin angiotensin-aldosterone system, originally conceived as an axis of sodium and fluid metabolism in inflammation, thrombosis and cardiac and smooth muscle hypertrophy, is a major factor in disease progression for conditions as diverse as hypertension, heart failure, coronary artery disease and diabetes. Interruption of angiotensin II generation and bradykinin degradation by ACE inhibition is a major therapeutic advance in the management of these diseases. NEP metabolizes both bradykinin and the natriuretic peptides (atrial natriuretic peptide, brain natriuretic peptide, c-type natriuretic peptide and adrenomedullin). These peptides counter the adverse effects of angiotensin II by their vasodilator, natriuretic, diuretic and autonomic neural actions; by their antitrophic effects; and by suppressing plasma renin activity. These two systems can be considered key components of a cardiorenal axis that maintains blood pressure and cardiopulmonary blood volume within a stable range. This balance is compromised in the setting of heart failure and primary hypertension. The combination of ACE and NEP inhibition should augment the beneficial hemodynamic and tissue effects of bradykinin and the natriuretic peptides. Vasopeptidase inhibition, therefore, is a novel approach to cardiovascular therapy, with implications for hypertension, heart failure, renal function and ischemic heart disease. PMID- 11875588 TI - Detection of coronary microvascular disease by means of cardiac scintigraphy. AB - A 76-year-old woman strictly defined as having cardiac syndrome X underwent cardiac scintigraphies. A reversible perfusion abnormality was identified by (201)thallium in the inferior segment of the left ventricle. (123)iodine meta iodo-benzyl-guanigine uptake showed extremely diminished uptake in the inferior segment of the myocardium. (123)iodine labelled beta-methyl-iodophenyl pentadecanoic acid myocardial single photon emission computed tomography showed decreased uptake of the inferior segment in the early image, whereas the delayed images revealed significant fill-in of the tracer in the inferior segment of the myocardium. These findings suggest that coronary microvascular dysfunction causes regional myocardial ischemia, resulting in metabolic and sympathetic abnormality. PMID- 11875590 TI - Coronary artery-posterior interventricular vein fistula after endomyocardial biopsy in a heart transplant patient. PMID- 11875589 TI - Acute myocardial infarction late after Mustard procedure for dextrotransposition of the great arteries. AB - Systemic right ventricular dysfunction has been closely linked to late mortality and sudden cardiac death in patients with Mustard procedure for dextrotransposition of the great arteries. Two young patients with dextrotransposition of the great arteries late after Mustard procedure who presented with acute transmural myocardial infarction and sudden cardiac death (one patient) without prior exertional angina or causative coronary abnormalities are reported. It is surmised that acute coronary emboli originating from a severely dilated, hypocontractile systemic ventricle were the cause of transmural myocardial infarction. This phenomenon may be an important and as yet unrecognized factor in late morbidity and mortality in such patients. PMID- 11875591 TI - A framework for (health) reform. PMID- 11875592 TI - What are we going to do with you? Gastroenterology service providers' perceptions of "difficult to manage" IBD patients. AB - BACKGROUND: This paper reports the results of a systematic survey of members of a clinical gastroenterology service to determine their perceptions of patients with inflammatory bowel disease (IBD) who were deemed to function poorly and were difficult to manage clinically. OBJECTIVES: To assess objectively the defining characteristics of this perceived subgroup of patients who are encountered in virtually all gastroenterology services. METHODS: A sample of gastroenterologists and gastrointestinal surgeons (n=10), as well as gastrointestinal nurses (n=19), was surveyed regarding their beliefs about the characteristics of patients with IBD who they judged to be extremely "difficult to manage". A survey was developed to assess patient characteristics (eg, symptom presentation, narcotic over reliance, interpersonal behaviour and illness behaviour) and the emotional impact that this perceived patient group has on individual staff members as well as on the functioning of the gastrointestinal team. RESULTS: The data indicated that patients with IBD who were perceived to be poorly functioning were viewed to have high levels of dysfunctional behaviour. In particular, negative behaviours (eg, manipulative interpersonal behaviours and excessive illness behaviours) were noted. Not only were these categories of behaviours high in frequency, but survey participants also rated these categories of behaviour to be highly distinct from those of typical patients with IBD. Moreover, this perceived patient group was reported to have a negative impact on individual staff and on the gastrointestinal team, and participants confirmed that they experience significant frustration and hostility when they work with these patients. CONCLUSIONS: These data, if replicated, confirm the general clinical opinion that a small subgroup of "difficult to manage" and poorly functioning patients with IBD exists. These patients appear to differ from typical patients with IBD in interpersonal characteristics more than in medical characteristics. If follow-up research, which is currently underway by the authors' group, shows that groups of poorly functioning gastrointestinal patients and typical gastrointestinal patients actually differ in measures of illness behaviour, then novel treatment approaches to improve the clinical services that are provided to these patients can be developed. PMID- 11875593 TI - Complementary practitioners' views of treatment for inflammatory bowel disease. AB - A substantial number of patients with inflammatory bowel disease use complementary therapies to manage their disease, including chiropractic and herbal therapies. The objective of this study was to explore whether providers of these therapies see patients with inflammatory bowel disease and recommend therapies, and to determine their opinions about the treatments that they recommend. The study sample comprised 66 chiropractors, 19 pharmacists, 16 herbalists and 15 health food store employees in Calgary, Alberta. A structured questionnaire containing two patient scenarios (a patient with active ulcerative colitis and a patient with inactive Crohn's disease) was completed either by an in-person interview or by a mailed questionnaire. Most respondents had seen patients with ulcerative colitis, and at least 80% of each group except pharmacists (only 10%) would treat these patients or recommend treatment. Almost all chiropractors used spinal manipulation, whereas herbalists and health food store employees suggested a wide range of different treatments. Chiropractors rated their treatment as moderately effective; herbalists and health food store employees viewed their recommendations as very effective. The results with respect to the second scenario were very similar. The wide range of treatment recommendations by practitioners, who differ greatly in terms of skills, knowledge and experience, has important implications for physician-patient communication, information provision and education regarding complementary and alternative therapies. PMID- 11875594 TI - Cancer of the pancreas: the best image for early detection--CT, MRI, PET or US? AB - Pancreatic cancer has a poor prognosis, and the best chance for survival is to diagnose the tumour at an early stage. Abdominal ultrasound, computed tomography, magnetic resonance imaging and endoscopic retrograde cholangiopancreatography are the most commonly used radiological techniques for imaging the pancreas. The diagnostic evaluation should be tailored to the individual patient. Dual-phase helical computed tomography and magnetic resonance imaging have similar accuracies for detecting and staging pancreatic adenocarcinoma. Sonography results are highly dependent on the skill and persistence of the operator. No radiological examination is very sensitive at visualizing small metastases in the lymph nodes and peritoneum, or on the surface of the liver. Thus, it is difficult to establish with certainty whether a tumour is resectable. Another major challenge is to differentiate cancer from an inflammatory mass in chronic pancreatitis. Functional imaging (using positron emission tomography with fluorodeoxyglucose) may be helpful, especially if the images are fused with those of computed tomography or magnetic resonance imaging. The diagnostic accuracies, applications and limitations of the various modalities are discussed. PMID- 11875595 TI - Identifying contraindications to resection in patients with pancreatic carcinoma: the role of endoscopic ultrasound. AB - OBJECTIVE: To present recently published material comparing the performance of endosonography relative to other imaging modalities when evaluating the patient with a suspected or known pancreas carcinoma. METHODS: Medline was searched using the terms "endosonography" and "pancreas neoplasms". References from retrieved papers were reviewed to identify other reports. Emphasis was placed on peer reviewed material published within the past three years that included comparison with other imaging modalities. RESULTS: Despite advances in cross-sectional imaging modalities, endosonography remains the most sensitive and specific method for identifying pancreatic mass lesions. The resectability of pancreatic carcinoma is best determined with dual-phase helical computed tomography, although endosonography may be slightly more accurate for lymph node assessment. Endoscopic ultrasound-guided fine needle aspiration biopsy has a high sensitivity (93%) and specificity (100%) when used in patients with masses in whom pancreatic cancer is suspected but prior biopsies have been negative. CONCLUSIONS: Endosonography helps in the diagnosis of pancreatic neoplasms through definitive inclusion or exclusion of a mass lesion as well as biopsy confirmation of malignancy. The role of endosonography in the determination of resectability has been eclipsed by dual-phase helical computed tomography. However, endoscopic ultrasound with fine needle aspiration of nonperitumoral lymph nodes may identify advanced disease with sufficient frequency to justify its routine use in patients with lesions that are thought to be resectable based on helical computed tomography. PMID- 11875596 TI - Pancreatic cancer: who benefits from curative resection? AB - Surgical resection is the only chance for cure of pancreatic cancer. Unfortunately, the majority of patients have grossly unresectable disease. Patients with stage I or II disease according to the criteria of the International Union Against Cancer (UICC) should be considered for potentially curative surgery. The goal of surgery is to remove the entire tumour with no residual disease (oncological R0 resection), which requires extensive resection of the surrounding tissues. Even if lymph nodes are histologically free of disease, molecular biological techniques reveal infiltration with cancer cells in 50% of cases. Therefore, extensive local resection combined with radical resection of lymphatic tissue, including lymph nodes around the head of the pancreas, retroperitoneal tissue and neural plexus around the great vessels, affords a longer median survival time than standard resection alone. Even patients with UICC stage III disease can undergo aggressive surgical treatment, but their chances for long term survival are low. Some patients develop severe diarrhea after circumferential removal of nerve tissue around the superior mesenteric artery. Adjuvant radiochemotherapy also provides a modest prolongation of survival. Despite these advances, the prognosis for pancreatic cancer is still poor, and spread of tumour within the peritoneum and to the liver is common postoperatively. PMID- 11875597 TI - Pancreatic cancer: what the oncologist can offer for palliation. AB - Because pancreatic cancer has a poor survival rate and only 20% of patients present with potentially resectable disease, a key goal of therapy is to provide palliation. The poor medical condition of many patients interferes with their ability to tolerate traditional chemotherapy. Recently, however, a nucleoside analogue, gemcitabine, has been developed. This drug is more effective than 5 fluorouracil (5-FU), can be used in patients who fail to respond to 5-FU and has only modest toxicity. Combination therapies including gemcitabine and other agents are being tested. Local radiotherapy seems to provide pain relief, but gastrointestinal toxicity is significant. The effect of combined modality therapy (5-FU with radiotherapy) on survival is unclear, and it does not prevent local disease progression. Some novel biological agents, including angiogenesis inhibitors, matrix metalloproteinase inhibitors, antisense compounds, inhibitors of cell signalling such as epidermal growth factor and vascular endothelial growth factor, and inhibitors of oncogene activation, are undergoing phase II and III trials in patients with pancreatic cancer. Among the most promising are farnesyl protein transferase inhibitors, which modulate K-ras function. Such an approach is promising for the treatment of pancreatic cancer because this tumour frequently exhibits mutation of the ras gene. PMID- 11875598 TI - Bugs, TNF-alpha and IBD: more fuel for the fire. PMID- 11875599 TI - Esophageal varices: TIPS or ligation. What is your final answer? PMID- 11875601 TI - Hypertension/preeclampsia. PMID- 11875602 TI - Manipulation of papillary muscle cyclic nucleotides during anoxia-reoxygenation: effects on contractility. AB - Endogenous nitric oxide and the accompanying cGMP formation has been postulated to play a role in the pathophysiology of myocardial anoxia-reoxygenation. A direct relationship between cGMP and the alterations observed in contractility under these conditions has never been demonstrated. In this study, cGMP in rat papillary muscles during anoxia and reoxygenation was correlated with mechanical function. Isolated papillary muscles were stimulated continuously and made anoxic for 40 min after a 2-hour stabilisation period. Anoxia caused an abbreviated contraction curve by decreasing the maximum contraction strength, time to peak contraction and relaxation time, accompanied by a significant decrease in tissue cGMP and cAMP levels (controls: 29.09 +/- 1.62 and 568.8 +/- 35.65; anoxia:16.62 +/- 1.51 and 403.3 +/- 30.19 pmoles/gww), which partially returned to pre-anoxic values upon reoxygenation. cGMp levels were significantly elevated by addition of 8-Br-cGMP (a cGMP analogue), but this elevation (154.4 +/- 20.89 pmoles/gww), had no effects on the contractility pattern of muscles during normoxia, anoxia or reoxygenation, suggesting that in isolated ventricular muscle, cGMP levels play a minor role in regulating muscle contractility. PMID- 11875603 TI - Maternal deaths due to pre-existing cardiac disease. AB - Heart disease in pregnancy is an uncommon problem in the developed world, but reaches a high prevalence in poor countries. In South Africa 0.65% of all pregnant women have heart disease, and there is an unacceptably high morbidity and mortality rate (9.5% ). Rheumatic heart disease accounts for most of this mortality, mitral stenosis being the commonest lesion. In April 2000 the National Committee on Confidential Enquiries into Maternal Deaths (NCCEMD) reported that close to half of non-obstetric maternal deaths in South Africa were due to cardiac disease. Several preventable factors were identified that precipitated decompensation and could have accounted for this high mortality. Among them, lack of adequate antenatal evaluation, uncontrolled fluid infusion, failure to identify the patient at risk, and failure to recognise the risk of autotransfusion in the postpartum phase, were contributing factors. This report of the problems seeks to address ways in which these difficulties may be rectified. PMID- 11875604 TI - Acute myocarditis in a patient with eosinophilia and pulmonary infiltrates. AB - This is a case of nectrotising myopericarditis complicated with an acute apical left ventricular aneurysm in a young patient with the history of bronchial asthma and recurrent eosinophilia. Prompt clinical improvement coinciding with normalisation of the blood count in response to steroids was observed. Possible aetiology of hypereosinophilia is discussed. PMID- 11875606 TI - Hepatitis C virus (HCV)-related cryoglobulinemia after liver transplantation for HCV cirrhosis. AB - The aim of this study was to analyze the clinical impact of hepatitis C virus (HCV)-related cryoglobulinemia in patients that had received liver transplants after HCV cirrhosis. Thirty patients who had received transplants between 1990 and 1996 for HCV cirrhosis and who had a follow-up longer than 1 year were studied. Serum HCV RNA levels, HCV genotype, cryoglobulinemia, rheumatoid factor, serum C3 and C4, IgA, IgG, IgM levels, liver tests, and liver histology were studied 30 +/- 16 months post-transplant. Cryoglobulinemia was found in 9 of 30 patients (30.0%) and was symptomatic in 4 of the 9 cases (glomerulonephritis, 1 case; palpable purpura, 3 cases). Age, sex distribution, alanine aminotransferase (ALAT) activity, and Knodell score did not differ, whether cryoglobulinemia was present or not. Rheumatoid factor (209.5 +/- 70.4 IU/l vs 12.0 +/- 4.4 IU/l, P = 0.004) and IgM levels (3.2 +/- 0.5 g/l vs 1.6 +/- 0.9 g/l, P = 0.0001) were significantly higher, and C4 levels (0.16 +/- 0.16 g/l vs 0.30 +/- 0.10 g/l, P = 0.009) were significantly lower in patients with cryoglobulinemia. One patient died from cryoglobulin-related renal failure. We concluded that, after liver transplantation (LT) for HCV cirrhosis, cryoglobulinemia was frequent and often symptomatic. Cryoglobulinemia did not seem to be associated with more severe graft damage. Cryoglobulinemia-associated morbidity must be taken into account in the management of post-transplant HCV infection. PMID- 11875607 TI - Risk factors for delayed graft function after renal transplantation and their significance for long-term clinical outcome. AB - Abstract. Delayed graft function (DGF) remains a grieving complication after renal transplantation. In this study, we examined various factors related to organ donation, transport, and transplantation for their influence on the incidence of DGF and on long-term prognosis. The incidence of DGF, renal function after 5 years, and allograft survival were analyzed in 200 kidneys transplanted in Dusseldorf as well as in 193 partner kidneys transplanted at 43 other centers. The main risk factors for DGF were donor age, cold ischemia time (CIT) and organ shipment. DGF itself, as well as donor age, influenced the long term prognosis. A significant relationship between the partner organs regarding clinical outcome was demonstrated. Non-immunological factors strongly influence the clinical results after renal transplantation. Organs of older donors have a limited long term prognosis. To minimize additional risks, prevention of DGF, especially by reducing CIT, should be regarded as of paramount importance. PMID- 11875608 TI - Anionic polysaccharides. A class of substances with hepatoprotective and antiadhesive properties in rat liver preservation. AB - In liver preservation, the substitution of the anion Cl(-) by lactobionic acid (LB) prevents reperfusion edema and extends the preservation time for human livers. We studied the effect of compounds that are structurally related to lactobionic acid: anionic polycarbohydrates (sulfated anionic polysaccharide, SAP, and pentosan polysulfate, PPS) on liver function and leukocyte-endothelial cell interaction in isolated perfusion and liver transplant models. Rat livers, cold-stored (24 h) in a Cl(-) -containing control solution, became edematous during 1 h of reperfusion. Substitution of Cl(-) by either LB, SAP, or PPS decreased reperfusion edema in a Cl(-) concentration-dependent fashion. Reperfusion edema was abolished completely after preservation in 100 mM SAP solution or PPS solution. Also hepatic lactic dehydrogenase (LDH) and aspartate aminotransferase (ASAT) release was lowest after preservation in those solutions. After preservation in LB or anionic polycarbohydrate solutions, portal venous resistance was significantly higher than after preservation in Cl(-)-containing control solution. Capillary blood flow was 391 +/- 83 pl/s and 398 +/- 174 pl/s after preservation in SAP solution (SAPs) and PPSs, and 803 +/- 117 pl/s and 641 +/- 219 pl/s after preservation in LB or Cl(-)-containing control solution. The number of leukocytes sticking to the vascular wall was lower ( P < 0.05) after preservation in SAPs or PPSs (109 +/- 31 cells/mm(2) and 108 +/- 60 cells/mm(2), respectively), when compared with preservation in Cl(-)-containing control or LB solutions (429 +/- 63 cells/mm(2) and 277 +/- 59 cells/mm(2)). In rat liver preservation, anionic polysaccharides are antiedematous compounds, with a higher potency than LB and additional antiadhesive properties. PMID- 11875609 TI - The value of switching from cyclosporine to tacrolimus in the treatment of refractory acute rejection and obliterative bronchiolitis after lung transplantation. AB - Standard cyclosporine-based immunosuppression is ineffective in the treatment of refractory acute rejection (RAR) and obliterative bronchiolitis (OB) that follows lung transplantation. The aim of this study was to evaluate the results of switching from cyclosporine to tacrolimus in the treatment of these situations. Nineteen patients entered the study. The indication for switching was OB in 11 patients and RAR in 8. Mean age was 41.3 +/- 13.1 years. In patients with RAR, the number of acute rejections was 1.5 +/- 0.7 and there were zero episodes per patient per 100 days before and after switching, respectively ( P = 0.02). There was no significant reduction of the decline of forced expiratory volume (FEV(1)) within 6 months after switching in patients with OB. We conclude that the conversion from cyclosporine to tacrolimus was associated with favourable results in the treatment of RAR. Further studies are required to assess the influence of this approach in the treatment of OB. PMID- 11875610 TI - The importance of orthotopic liver transplantation in acute hepatic failure. AB - Selection of patients with acute hepatic failure for liver transplantation remains difficult, and there is no definite proof of a survival effect. We therefore did a retrospective study in 75 consecutive patients referred over a 12 year period. In two-thirds we identified a cause, mostly viruses or drugs. Patients were grouped by the Clichy and King's College criteria. In 20 there was no indication for transplantation. Of the 5 with autoimmune hepatitis, 3 died, significantly differing from the other 15 ( P = 0.009). The remaining 55 met our criteria, except 1. All 9 patients with absolute contraindications died. Of the 46 enlisted, 7 died without transplantation. One-year survival after transplantation was 69%, compared with 58% by "intention to treat." For patients enlisted, transplantation reduced mortality by 78% ( P = 0.069). The Clichy and King's College criteria reliably predict survival without transplantation, except in autoimmune hepatitis. Our study strongly suggests that transplantation improves survival. PMID- 11875611 TI - Outcome of orthotopic liver transplantation in autoimmune hepatitis according to subtypes. AB - The relevance of autoimmune hepatitis (AIH) classification for clinical purposes is controversial. We analyzed the outcome after orthotopic liver transplantation (OLT) of nine type I and seven type II AIH patients. Type II patients had a significantly higher incidence of cirrhosis at the time of diagnosis, more resistance to steroid therapy, and a higher Child-Pugh score at the time of OLT. OLT was performed in emergency in three type II patients and electively in all type I patients. Four type II and one type I patients died in the postoperative period. There was no difference regarding the incidence of post-OLT infection and rejection between the two types. No recurrence of AIH was observed. The 6-year actuarial survival rates for type I and type II patients were 76% and 43%, respectively. Type II AIH patients who have a poor response to medical therapy should be considered for OLT with a shortened delay. PMID- 11875612 TI - Double bone marrow transplantation for severe aplastic anemia after orthotopic liver transplantation: implications for clinical management and immune tolerance. AB - A 2-year-old boy underwent liver transplantation for fulminant hepatic failure of unknown cause. Four months later the child developed severe aplastic anemia. Allogeneic bone marrow transplantation (BMT) was performed using marrow from his 14-month-old HLA-identical sister. Severe aplastic anemia recurred 2.5 months later. After reconditioning a second BMT was performed using the same donor. Tapering of immunosuppression 2 years after BMT led to biopsy-confirmed rejection of the liver. Therapy with high-dose corticosteroids and an increase in cyclosporine A medication readily reversed rejection and a low-dose immunosuppression reflected by cyclosporine trough levels less than 50 ng/ml has been maintained since. Eight years later the boy is in excellent health with both bone marrow and liver functioning perfectly. In summary, this case demonstrates that even recurrent severe aplastic anemia after OLT can be cured by BMT, and that a transplanted liver can tolerate a double conditioning regimen without problems. Tolerance towards the liver through BMT did not develop. PMID- 11875613 TI - Caliceal fistula in kidney transplantation. The role of magnetic resonance imaging. AB - Caliceal fistula is a rare complication of renal transplantation, which often raises some diagnostic problems. We report the case of a patient in which this complication occurred and in whom the diagnosis could be clearly demonstrated by using magnetic resonance imaging (MRI). On the T1-weighted images, a perirenal collection was depicted by a low signal intensity. On T2-weighted images, the collection appeared with a high signal intensity, and a linear hyperintensity was observed on the internal graft's labium at the level of the inferior pole corresponding to a caliceal fistula arising from the lower pole of the graft. In this setting, the use of MRI is compared with the other diagnostic techniques (sonography, CT scan, nephrogram, scintigraphy). MRI constitutes a progress in imaging of the renal graft by its high definition and the lack of nephrotoxicity. Its place remains, however, to be more precisely defined in the evaluation of a renal graft's complications. PMID- 11875614 TI - Reversal of oxalosis cardiomyopathy after combined liver and kidney transplantation. AB - Few data have been published on the course of oxalosis cardiomyopathy after combined liver and kidney transplantation in hyperoxaluria patients with myocardial involvement. We report the case of a primary hyperoxaluria type 1 patient with renal failure who developed end-stage cardiomyopathy. Left venticulography showed severe diffuse hypokinesia and left ventricular ejection fraction was calculated at 12%. Endomyocardial biopsy demonstrated platelike calcium oxalate crystals within the myocardium and the connective tissue, and mild perivascular fibrosis. The patient was first considered for combined liver heart-kidney transplantation, but as his cardiac function improved slightly with an intensive dialysis program, combined liver and kidney transplantation was performed. Normal cardiac function was demonstrated at 1-year follow-up, and comparative endomyocardial biopsy showed regression of the myocardial oxalate deposits. This case adds stronger clinical, hemodynamic, and histopathological evidence that severe oxalosis cardiomyopathy may be reversed after combined liver and kidney transplantation. PMID- 11875615 TI - Hepatic artery pseudoaneurysm after liver transplantation. A result of transhepatic biliary drainage for primary sclerosing cholangitis. AB - Hepatic artery pseudoaneurysm (HAP) is a rare but often life-threatening complication of liver transplantation. Treatment is usually ligation, revascularization, or retransplantation. We report a patient suffering from primary sclerosing cholangitis who required transhepatic percutaneous biliary drainage. Following orthotopic liver transplantion (OLT), he developed HAP, which was successfully embolized angiographically. We discuss the association between transhepatic biliary drainage and the development of HAP after OLT, and the possibilities for angiographic intervention in these cases. PMID- 11875617 TI - ESOT - Novartis Study Grant: Call for Applications, Guidelines and Application form. PMID- 11875616 TI - Outcome of an AB0-incompatible renal transplant without splenectomy. PMID- 11875618 TI - Activation of the hexose monophosphate shunt in rat type II pneumocytes as an early marker of oxidative stress caused by cobalt particles. AB - Cobalt metal (mCo) and hard metal, a mixture of cobalt and tungsten carbide (CoWC), are cytotoxic for alveolar macrophages and alveolar type II cells (AT II). Although the exact mechanisms of toxicity are not entirely elucidated, evidence exists for an oxidant-mediated toxicity. In this study, we exposed primary cultures of rat AT-II, in vitro, to different forms of cobalt (mCo particles, CoWC particles, CoCl(2)) and assessed changes in the activity of the hexose monophosphate shunt (HMS). Activation of the HMS occurs as an early response to (per)oxidative stress. Cobalt metal-containing particles (mCo and CoWC) when freshly immersed in medium, lead to an early concentration-dependent stimulation of the HMS in rat AT-II. The maximum stimulations of HMS (reached after 90 min) were 2.0 +/-1.2, 2.9+/-0.4, 3.3 +/-1.6 and 4.0+/-0.4 fold-increases for 15, 75, 300 and 1200 microg mCo/well, respectively. The observed time course of the activation by mCo particles clearly differed from that caused by paraquat (10(-5 )M), which is known to produce activated oxygen species after cyclic oxidation-reduction reactions. The comparable effect of peroxides (H2O2 and t butyl hydroperoxide) on HMS and the inhibitory effects of catalase on the mCo induced stimulation of the HMS strongly suggest the production of peroxides by freshly immersed mCo particles. However, we were not able to show a simple relationship between the stimulation of the HMS and the subsequent cell damage. PMID- 11875619 TI - Cadmium exposure decreases androgen-dependent metabolism of acetohexamide in liver microsomes of male rats through its testicular toxicity. AB - Administration of cadmium (Cd) at a dose of 1.23 mg/kg (2.0 mg/kg as CdCl(2)) markedly decreased the activity of an enzyme (acetohexamide reductase) catalysing the ketone-reduction of acetohexamide, an oral antidiabetic drug, in liver microsomes of male rats. However, the decreased enzyme activity was increased by repeated treatment with testosterone propionate (TP). When male rats were castrated and TP was given to the castrated ones, a similar decrease and increase, as described above, were observed in the microsomal enzyme activity. Cd exposure to male rats induced haemorrhage and atrophy of the testes and significantly diminished serum testosterone levels. There was no possibility that Cd accumulated in liver microsomes of male rats causing direct inhibition of the microsomal enzyme activity. We conclude that Cd exposure decreases androgen dependent metabolism of acetohexamide in liver microsomes of male rats through its testicular toxicity. Cd exposure had no effect on acetohexamide reductase activity in liver cytosol of male rats. PMID- 11875620 TI - Critical analysis of biomonitoring endpoints for measuring exposure to polymeric diphenyl-methane-4,4'-diisocyanate (MDI) in rats: a comparison of markers of exposure and markers of effect. AB - The object of this study was to compare the relative sensitivity of markers of exposure and effects in the lung of rats exposed to polymeric diphenyl-methane 4,4'-diisocyanate (pMDI) aerosol. Rats were repeatedly exposed to 12.9 mg pMDI/m(3) (6 h/day, 5 days/week for 14 days; exposure was from days 0--17 followed by a post-exposure period to day 35). Markers of exposure were determined in bronchoalveolar lavage (BAL), blood (haemoglobin, plasma proteins), and urine on days 1, 4, 11, 18, 21, 28 and 35. Markers of effects were determined at the same time points and focused on changes in BAL constituents. In BAL, a maximum increase of total protein occurred following the first exposure and levelled off subsequently whilst BAL cell-related endpoints increased in a time dependent manner. The kinetics of formation and elimination of adducts differed appreciably from one dosimeter to another. Whilst haemoglobin adducts were integrated by the cumulative exposures, the incremental yield of adduct formation appeared to be dependent on pulmonary as well as yet unknown erythrocyte-related factors. Plasma protein adducts attained a plateau after 1 week of exposure. MDI related metabolite levels in urine did not show any time-dependent changes during the entire exposure period and declined rapidly during the post-exposure period. Thus, the kinetics of the fractional loading and clearance of pulmonary and extrapulmonary dosimeters did not parallel each other, nor was there a clear correlation with the markers of effects. In summary, it is concluded that biomonitoring is a powerful tool for the comparative dosimetry of well-defined exposure regimens. However, especially for irritant agents demonstrating portal of-entry effects, markers related to 'total body burden' may not necessarily predict the absence or presence of local responses occurring within the target organ, namely the lung. In all compartments, dosimeters related to higher oligomers of MDI demonstrated low bioavailability, i.e. their recovery was appreciably lower than expected. PMID- 11875622 TI - Inhibition and recovery of maternal and fetal cholinesterase enzymes following a single oral dose of chlorpyrifos in rats. AB - Pregnant Sprague-Dawley rats (14-18 days of gestation) were treated with a single dose of 50 mg/kg (61% of oral LD50 in female rats) of chlorpyrifos ( 0,0-diethyl- 0-3,5,6-trichloro-2-pyridyl phosphorothioate) by oral gavage. Animals treated on day 18 of gestation were sacrificed at 1, 2, 4, 12 h after dosing. Animals treated on days 17, 16, 15, and 14 of gestation were sacrificed at 24, 48, 72, and 96 h after dosing, respectively. Maternal and fetal brain acetylcholinesterase (AchE) and plasma butyrylcholinesterase (BuChE) activities were significantly inhibited 1 h after treatment. Activity of fetal brain AChE and plasma BuChE recovered faster than that of the maternal enzymes. Peak inhibition of maternal spinal cord AChE and BuChE activities occurred 2 h and 1 h after dosing, respectively. Maternal spinal cord BuChE activity was totally recovered by 96 h compared to the partial recovery of spinal cord AChE activity. Maternal liver BuChE activity was significantly decreased within 1 h of dosing. The individual molecular forms (10S and 4S) of maternal and fetal brain AChE and BuChE activities were significantly decreased 1 h after treatment. Recovery of both forms of fetal brain AChE activity was much faster than the maternal forms. Activity of the 10S form of maternal control brain AChE was significantly higher than in the fetus control. The rapid recovery of cholinesterase enzymes in the fetus is attributed to the de novo synthesis of AChE enzymes in the fetus compared to the mother. PMID- 11875621 TI - Transplacental transfer of the phytoestrogen daidzein in DA/Han rats. AB - Disposition and transplacental transfer of the phytoestrogen daidzein was studied in pregnant DA/Han rats on day 18 of gestation. Daidzein concentrations were determined by HPLC in maternal blood, maternal organs (liver, kidney, uterus), placenta and fetuses (liver and residual tissues) at specific times (5, 10, 20, 40 and 120 min) after intravenous administration of 10 mg/kg body weight. Early after injection, the majority of circulating daidzein was still in the aglycone form; at later time points the majority consisted of conjugates. The initially high isoflavone concentration in maternal plasma (about 25 microg/ml at 5 min) decreased rapidly within the first hour, and after 2 h total daidzein was below 1 microg/ml. Despite its efficient conjugation, daidzein was rapidly distributed in the organism: peak concentrations were attained 10 min after intravenous administration in all tissues analysed, with mean values of about 31 microg/g in maternal liver, 13 microg/g in kidneys and 5 microg/g in the uterus. Placenta contained about one-tenth the hepatic daidzein concentration, and fetal liver about 1/30 the peak concentration of maternal liver (i.e. 1.3 microg/g, which is one-third the placental concentration). Daidzein levels in tissues then declined in parallel with those in maternal blood. The data show that daidzein is transferred across the placenta of DA/Han rats to fetuses. This is indicative of a rapid transfer from the mother to the fetus, but also that efficient hepatic extraction of daidzein from the maternal blood occurs. Since dietary phytoestrogens account for a significant proportion of human exposure to potential endocrine modulators, and since the placenta does not represent a barrier to daidzein or related estrogenic isoflavones, the consequences of these exposures early in life should be examined and monitored carefully. PMID- 11875623 TI - Functional and morphological effects of styrene on the auditory system of the rat. AB - The effect of inhaled styrene on the structure and function of the auditory organ of the male Wistar rat was studied. The animals were exposed either to 600, 300 or 100 ppm styrene (12 h/day, 5 days/week, for 4 weeks). Auditory sensitivity was tested prior to and after the exposure by auditory brain stem audiometry (ABR) at frequencies of 1.0, 2.0, 4.0 and 8.0 kHz. Inner ear morphological changes were studied by light- and electron-microscopy. Exposure to 600 ppm styrene caused a 3 dB hearing loss only at the highest test frequency (8 kHz). Quantitative morphological analysis of cochlear hair cells (cytocochleograms) showed that 600 ppm styrene caused a severe outer hair cell (OHC) loss particularly in the third OHC row of the upper basal and lower middle coil. The inner hair cells were usually intact. Exposure to lower styrene concentrations (100 and 300 ppm) caused no unequivocal functional deficit or hair cell damage. We conclude that there appears to be a concentration threshold for styrene ototoxicity in rats (between 300 and 600 ppm). PMID- 11875624 TI - Humic acid extracted from Blackfoot disease-endemic well water induces adipocyte differentiation of C3H10T1/2 fibroblast cells: a possible mechanism leading to atherosclerotic-like plaque in Blackfoot disease. AB - A unique peripheral vascular disease named "Blackfoot disease" (BFD) is endemic on the southwest coast of Taiwan. Clinically, the signs and symptoms of BFD are similar to those of arteriosclerosis and Buerger disease. Humic acid has been proposed as a causative factor in BFD; however, the relationship between humic acid and atherosclerotic-like plaque associated with BFD remains unclear. In this study, we investigated the effects of humic acid extracted from Blackfoot disease endemic well water (BFD-HA) on cultured C3H10T1/2 fibroblasts, a murine embryonic cell line. Our present data demonstrate that C3H10T1/2 cells were arrested at the G1 phase and subsequently differentiated to adipocytes after treatment with BFD HA. The adipocyte differentiation of C3H10T1/2 cells induced by BFD-HA was also accompanied with increased glycosaminoglycan production. These results suggest that a large lipid accumulation of arterial blood vessels in BFD patients may be linked in part to the adipocyte differentiation of vascular fibroblasts induced by BFD-HA. PMID- 11875627 TI - 10th International Course on the Safety Assessment of Medicines Hyeres, France September 8-13, 2002. PMID- 11875625 TI - Cytotoxic and genotoxic effects of fumonisin B1 on rabbit kidney RK13 cell line. AB - Fumonisins, mycotoxins produced by certain strains of Fusarium moniliforme, could induce various diseases in animals and are suspected human carcinogens. Fumonisin B1 (FB1), the most commonly found fumonisin, has been characterised as a tumour initiator and a tumour promoter, a mitogen and an anti-proliferative agent. In this study we examined the cytotoxicity and genotoxicity of FB1 in rabbit kidney RK13 cells. To evaluate the effects of FB1 on survival of this cell line we analysed cell viability, membrane integrity, DNA fragmentation and overall morphology of the cells. The genotoxic potential of FB1 was estimated by monitoring the ability of this mycotoxin to induce micronuclei in RK13 cells. Exposure to FB1 caused a significant increase in micronucleus frequency in a concentration- and in a time-dependent manner. Nanomolar concentrations of FB1 decreased cell viability after 24 h and even more so after 48 h of exposure. The morphological changes observed suggested that an increased number of RK13 cells were dying by the process of apoptosis. However, FB1 also induced impairments of cell and mitochondrial membrane integrity, as assessed by lactate dehydrogenase and glutamate dehydrogenase leakage. These results could imply that nanomolar concentrations of FB1 induced apoptosis, which subsequently may proceed to secondary necrosis. In summary, our observations suggest that FB1 is genotoxic and cytotoxic to RK13 cells. PMID- 11875628 TI - Time-dependent induction of anxiogenic-like effects after central infusion of urocortin or corticotropin-releasing factor in the rat. AB - RATIONALE: Corticotropin-releasing factor (CRF) and urocortin (Ucn) belong to the CRF-related family, share a high degree of structural homology and bind to CRF receptors. However, compared with CRF, Ucn was shown to display either weaker or similar anxiogenic-like effects in vivo. OBJECTIVE: To compare the anxiogenic like responses of rats injected intracerebroventricularly (ICV) with different doses of either rat/human CRF (r/hCRF) or rat Ucn (rUcn) at different intervals after injection. METHODS: Rats were tested on three validated paradigms of emotional behavior [i.e. elevated plus-maze (EPM), defensive withdrawal (DW) and conflict test (CT)] 5 and 30 min after treatment. RESULTS: In the EPM test only r/hCRF, but not rUcn, produced anxiogenic-like effects at the dose of 1.0 microg, when the peptides were injected 5 min before testing. At 30 min after injection, both peptides caused a significant reduction of open arms exploration, rUcn being effective at 0.01 microg. In the DW test both peptides were equally potent in decreasing the exploratory behavior and increasing the time spent in the chamber at the dose of 1.0 microg when tested 30 min after injection. In the CT both rUcn (0.25-1.0 microg) and r/hCRF (0.75-1.0 microg) decreased significantly the responding in the punished component. However, rUcn reduced food responding also in the unpunished component possibly due to its powerful anorectic activity. CONCLUSIONS: Comparison of anxiogenic-like activities of r/hCRF and rUcn at doses up to 1.0 microg revealed striking differential effects that depended on the time of testing after ICV peptide injection, and on the paradigm of anxiety used. These results suggest that the onset of r/hCRF and rUcn actions related to behavioral responses to anxiety is likely to depend on brain peptide-specific mechanisms including binding properties to CRF-receptors, differential distribution to specific functional brain sites and the distribution and effectiveness of binding-protein interactions. PMID- 11875629 TI - Effect of MS-153 on the development of behavioral sensitization to locomotion- and ataxia-inducing effects of phencyclidine. AB - RATIONALE: Repeated administration of phencyclidine (PCP) produces behavioral sensitization to PCP. Although the precise mechanism is unknown, glutamatergic neurotransmission seems to play an important role in the development of sensitization. OBJECTIVES: The present study examined whether a novel compound, MS-153 (( R)-(-)-5-methyl-1-nicotinyl-2-pyrazoline), which has an ability to enhance glutamate uptake and inhibit glutamate release, would block the development of behavioral sensitization to PCP. METHODS: For studying effects of MS-153, locomotor activity was measured by an infrared sensor and ataxia was measured by a rating scale. RESULTS: MS-153 (10 and 100 mg/kg) enhanced locomotion and ataxia induced by a single injection of PCP (7.5 mg/kg). Repeated administration of PCP (20 mg/kg, once in every day, for 5 days) developed sensitization to locomotion- and ataxia-inducing effects of PCP (7.5 mg/kg). MS 153 given 60 min and 120 min later of every PCP treatment blocked the development of behavioral sensitization to both locomotion- and ataxia-inducing effects of PCP. Co-administration of MS-153 with repeated saline treatment did not produce hypersensitivity to PCP. CONCLUSIONS: These results suggest that the attenuation of glutamatergic neural transmission enhances acute effects of PCP, in contrast, blocks the behavioral sensitization developed by repeated PCP treatment. Therefore, glutamatergic neural transmission plays an important role in the development of behavioral sensitization to PCP. PMID- 11875630 TI - Characterization of the discriminative stimulus effects of buprenorphine in pigeons. AB - RATIONALE: Buprenorphine is a low-efficacy mu opioid agonist that can reduce drug taking in opioid abusers; however, the mechanism by which buprenorphine modifies the actions of other drug taking and the consequences of repeated treatment with buprenorphine are not fully understood. OBJECTIVE: The purposes of this study were to evaluate the time- and dose-dependence of discriminative stimulus effects in pigeons receiving buprenorphine repeatedly and to examine possible interactions between buprenorphine and heroin. METHODS: Six pigeons discriminated between vehicle and 0.178 mg/kg buprenorphine while responding under an FR schedule for food. Substitution and drug combination studies characterized the potency and time course for buprenorphine, as well as interactions between buprenorphine and heroin. RESULTS: Stimulus control by buprenorphine was maintained throughout the study and was not changed by repeated daily dosing or by an acute injection of large doses of buprenorphine. Mu opioid agonists substituted for buprenorphine with the following order of potency: heroin > or = butorphanol > nalbuphine > or = morphine. Ketamine, enadoline, spiradoline, amphetamine and cocaine failed to substitute completely for buprenorphine. The discriminative stimulus effects of buprenorphine lasted 2-72 h, depending on dose, and naltrexone prevented but did not reverse the effects of buprenorphine. CONCLUSION: Despite a very long duration of action and apparent irreversibility, under these conditions in pigeons, buprenorphine does not modulate the discriminative stimulus effects of itself or heroin. Thus, simple agonism might account for the therapeutic effectiveness of buprenorphine in opioid abusers. PMID- 11875631 TI - Chronic inhibition of alpha4beta2 nicotinic receptors in the ventral hippocampus of rats: impacts on memory and nicotine response. AB - RATIONALE: Acute and chronic systemic nicotine administration has been shown to cause significant spatial memory improvement. The critical nicotinic receptor subtypes for this effect and their location are still being determined. Nicotinic receptors in the ventral hippocampus have been found to be critically involved in memory. Acute ventral hippocampal infusions of dihydro-beta-erythroidine (DHbetaE), an alpha4beta2 nicotinic receptor antagonist, impaired spatial memory of rats in the radial-arm maze. OBJECTIVES: The current study used chronic ventral hippocampal infusion of DHbetaE as a model of nicotinic receptor loss such as that which occurs in Alzheimer's disease. The therapeutic effect of systemic nicotine treatment in reversing the DHbetaE-induced memory impairment was determined. METHODS: Rats were pretrained to asymptotic levels of performance on the eight-arm radial maze. Then, they were implanted with bilateral infusion cannulae in the ventral hippocampus, through which 0, 33.3, or 100 microg/side/day of DHbetaE was continuously infused for 4 weeks. The rats were retested on the eight-arm maze throughout infusion period and after withdrawal, and the interaction of acute systemic nicotine injections on memory was tested. RESULTS: The higher (100 microg/side/day) but not the lower (33.3 microg/side/day) DHbetaE dose caused a significant spatial memory impairment. Acute systemic nicotine injections (0, 0.1, 0.2, and 0.4 mg/kg, subcutaneous) attenuated the memory impairing effects of 100 microg/side/day of DHbetaE. There was no significant effect on response latency with the chronic DHbetaE infusion. Acute systemic nicotine infusions did significantly speed responding, an effect which was reversed by chronic hippocampal infusions of DHbetaE. After withdrawal there were no significant lasting effects on choice accuracy or response latency. Wet-dog shakes were significantly elevated during chronic hippocampal DHbetaE administration with no effect during the withdrawal period. CONCLUSIONS: These results indicate that chronic inhibition of a subset of nicotinic receptors in the hippocampus results in a significant impairment in the spatial memory choice accuracy. The ability of nicotine to attenuate the impairment supports the development of nicotinic agonist therapy of syndromes, such as Alzheimer's disease, that involve a chronic decrease in the activity of the alpha4beta2 nicotinic receptors and memory impairment. PMID- 11875632 TI - Neuropeptide Y administration into the third ventricle does not increase sucrose or ethanol self-administration but does affect the cortical EEG and increases food intake. AB - RATIONALE: Several studies have provided indirect evidence that neuropeptide Y (NPY) may play a role in the regulation of ethanol consumption. However, the direct effects of central NPY administration on ethanol drinking are unclear. OBJECTIVE: This study examined the effects of NPY on ethanol, sucrose, and food consumption as well as its concomitant effects on the cortical EEG. METHODS: Wistar rats were implanted with cortical recording electrodes and a cannula in the third ventricle after using a sucrose substitution procedure to establish ethanol self-administration. NPY (0-15 microg/3.0 microl) was infused into the third ventricle prior to drinking sessions, when 10% ethanol (10E), 2% sucrose (2S), 0.5% sucrose (0.5S), or food were available. Behavior and cortical EEG were monitored during the sessions. RESULTS: NPY had no effect on the intake of 10E, 2S, or 0.5S, but NPY (15 microg/3.0 microl) significantly increased food intake. Under baseline drinking conditions, EEG power in the 6-8 Hz range was significantly greater when 2S was consumed compared to 10E. NPY decreased power in the 8-16 Hz range, decreased peak frequency in the 6-8 Hz range, and increased peak frequency in the 32-50 Hz range when 10E or 2S was available. CONCLUSIONS: These data suggest that NPY administration into the third ventricle preferentially regulates feeding compared to ethanol or sucrose drinking. In addition, since NPY significantly altered the cortical EEG in the absence of effects on ethanol and sucrose consumption, these data may indicate that NPY's cortical EEG effects are more related to its sedative or anxiolytic properties, rather than any effect on consumption. PMID- 11875633 TI - The contribution of the metabolite p-hydroxyamphetamine to the central actions of p-methoxyamphetamine. AB - RATIONALE: Para-methoxyamphetamine (PMA) is a substituted amphetamine that has been responsible for a number of fatalities in Australia and North America. Previous investigators have shown that p-hydroxyamphetamine (PHA), the primary metabolite of PMA, has effects on central neurotransmitter kinetics in vitro that are similar to those of the parent compound. In order to understand the role of PHA, it is necessary to determine both the in vivo actions and the concentrations achieved relative to those of PMA. OBJECTIVES: The effects of PHA and PMA on 5 hydroxytryptamine (5HT) and dopamine kinetics in brain were determined and the concentrations of each compound measured in blood and brain. METHODS: Animals were housed at 20-22C on a standard 12/12-h light/dark cycle. High speed chronoamperometry was used to compare the ability of PMA and PHA to alter 5HT and dopamine kinetics in the rat striatum in vivo. Concentrations of PHA and PMA in blood, whole brain and striatum were determined following a dose of PMA (10 mg/kg, IP.) using HPLC with fluorescence detection. RESULTS: PHA was more effective than PMA at evoking neurotransmitter release and inhibiting the uptake of dopamine. However, both compounds were approximately equipotent 5HT uptake inhibitors. PMA and PHA concentrations in whole brain and striatum peaked within 30 min of the administered dose, whereas blood concentrations of both compounds peaked 1 h after the dose. PHA concentrations in both blood and brain were consistently much lower than PMA concentrations. CONCLUSIONS: These data indicate that although PHA is more effective than PMA at altering 5HT and dopamine kinetics in vivo, it is unlikely to achieve sufficient brain concentrations to contribute to the central effects of PMA. PMID- 11875634 TI - Dopamine D(2) receptors and ingestive behavior: brainstem mediates inhibition of intraoral intake and accumbens mediates aversive taste behavior in male rats. AB - RATIONALE: One of the factors that terminate the ingestion of an intraorally infused solution of sucrose may be an increase in the perceived aversiveness of its taste. OBJECTIVES: We tested the hypothesis that dopamine D(2), as opposed to D(1), receptors in the brainstem or nucleus accumbens inhibit intraoral intake by enhancing the aversiveness of the taste of the infused solution. METHODS: Male rats were infused intraorally with a 2 M sucrose solution (1 ml/min) and intake and the display of gapes and chin rubs, i.e. taste-related aversive behavior, was measured. Gapes and chin rubs were also measured in rats during and 40 s after brief intraoral infusion (1 ml/min during 20 s) of a 0.3 mM solution of quinine HCl. The full D(1) receptor agonist dihydrexidine (0.1-3.0 mg/kg) and antagonist SCH-23390 (0.03-0.1 mg/kg), the D(2) receptor agonist quinpirole (0.3 mg/kg) and antagonist raclopride (1.7 mg/kg) were injected IP. Quinpirole (14-55 microg) and raclopride (5 microg) were also infused into the fourth brain ventricle. In addition, quinpirole (2 or 10 microg) was infused into the shell region of the nucleus accumbens. RESULTS: IP dihydrexidine and quinpirole inhibited the intraoral intake of sucrose and pretreatment with raclopride, but (in the case of dihydrexidine) not SCH-23390, attenuated this effect. Injection of quinpirole into the fourth ventricle produced raclopride-reversible inhibition of intraoral intake but did not stimulate the display of gapes and chin rubs. Infusion of quinpirole into the shell region of the nucleus accumbens had the opposite effects. The intake of sucrose was suppressed by the addition of quinine HCl but this suppression was unaffected by dopamine agonist or antagonist treatment. CONCLUSIONS: It is suggested that brainstem dopamine D(2) receptors mediate suppression of consummatory ingestive behavior and that D(2) receptors in the shell region of the nucleus accumbens mediate the display of gapes and chin rubs, but that neither of these D(2) receptor populations mediate the hedonic evaluation of taste. PMID- 11875635 TI - Sex and rat strain determine sensitivity to kappa opioid-induced antinociception. AB - RATIONALE: Recent studies indicate that sex and rodent strain are determinants of sensitivity to opioid-induced antinociception. OBJECTIVES: The present study examined the influence of sex and rat strain on kappa opioid-induced antinociception using a series of kappa opioids that vary in their relative effectiveness. METHODS: In a warm-water (50, 52 and 55C) tail-withdrawal procedure, the antinociceptive effects of kappa opioids were determined in male and female rats of the F344, Lewis and Sprague-Dawley (SD) strains. RESULTS: In both males and females of each strain, spiradoline produced high levels of antinociception across all nociceptive stimulus intensities, whereas U50,488 produced high levels only at the low and moderate nociceptive stimulus intensities. Sex differences in the potency and effectiveness of these kappa opioids were relatively small and not consistently obtained. Enadoline, bremazocine and nalorphine were less effective than spiradoline in producing antinociception, and at low and moderate nociceptive stimulus intensities these opioids were both more potent and effective in F344 and SD males than their female counterparts. In contrast, in Lewis rats, only bremazocine was more potent and effective in males. In combination tests, bremazocine shifted the spiradoline dose-effect curve leftward and/or upward in males and rightward in females (i.e., antagonized spiradoline). In contrast, in both males and females enadoline shifted the spiradoline dose-effect curve leftward and/or upward. CONCLUSIONS: These data indicate that kappa opioids were generally more potent and effective as antinociceptive agents in males than females. Similar to data obtained with micro opioids, the magnitude of these sex differences was generally larger with the less effective kappa opioids and determined, in part, by rat strain and nociceptive stimulus intensity. PMID- 11875636 TI - Effects of antipsychotic drugs on operant responding after acute and repeated administration. AB - RATIONALE: The current generation of atypical antipsychotic drugs represents an improvement over traditional ("typical") antipsychotics in many respects. However, a theoretical framework and adequate preclinical models have not yet been developed to predict or explain differences among the atypical antipsychotics, a necessary component of future development. OBJECTIVES: The purpose of the present set of experiments was to identify differences between the acute and subchronic effects of several atypical antipsychotic drugs and the typical antipsychotic haloperidol on operant responding in rats. METHODS: The effects of haloperidol and the atypical antipsychotics clozapine, olanzapine, risperidone, sertindole, quetiapine, remoxipride, and thioridazine were determined in rats trained to respond for food reward under a multiple fixed ratio 30/fixed interval 60 s schedule. A profile of the acute effects of each drug on response rates, response durations, and within-session effects were determined. Next, the dose of each drug that produced 75% suppression of response rates was administered for 16 consecutive days to determine whether or not tolerance would develop to the rate-suppressing effects of that dose. RESULTS: All drugs produced dose-related decreases in response rates. Only haloperidol and risperidone produced significant increases in response duration, while only haloperidol and remoxipride displayed within-session response decrements. Tolerance was evident for clozapine and to a lesser extent thioridazine. CONCLUSIONS: These results illustrate that the current generation of atypical antipsychotics are a heterogeneous group and that operant procedures may be useful for identifying differences preclinically. Specifically, clozapine appears to possess properties that distinguish it from other atypical antipsychotics, particularly after repeated dosing. PMID- 11875637 TI - A dose-finding study on the effects of branch chain amino acids on surrogate markers of brain dopamine function. AB - RATIONALE: We have previously shown in healthy volunteers that an amino acid mixture lacking tyrosine and phenylalanine reduces tyrosine availability to the brain and produces cognitive and neuroendocrine effects consistent with reduced dopamine function. This could provide a potential nutritional approach to disorders such as mania and schizophrenia, which are characterised by overactivity of dopamine pathways. The amino acid mixture we tested previously is unpalatable, whereas mixtures containing only branch chain amino acids can be made more palatable. However, the effects of such mixtures on dopamine function in humans have not been studied. OBJECTIVE: To assess the tolerability of different doses of branch chain amino acids and to measure their effects on neuroendocrine and cognitive measures sensitive to changes in dopamine function. METHODS: We used a randomised, double-blind, cross-over design in 12 healthy volunteers to assess the effect of single oral doses of 10 g, 30 g and 60 g branch chain amino acids on plasma prolactin and a test of spatial recognition memory RESULTS: The branch chain amino acids were well tolerated. The availability of tyrosine for brain catecholamine synthesis decreased in a dose related manner. As hypothesised, the drink increased both the plasma prolactin and the latency to respond on the spatial recognition memory task. CONCLUSIONS: A drink containing branch chain amino acids is well tolerated in healthy volunteers and produces effects consistent with lowered dopamine function. PMID- 11875638 TI - Pharmacological manipulations of the pedunculopontine tegmental nucleus in the rat reduce self-administration of both nicotine and cocaine. AB - RATIONALE: The pedunculopontine tegmental nucleus (PPTg) has been implicated in the self-administration of drugs, particularly nicotine, which acts directly through the PPTg in addition to targeting midbrain dopamine neurons. The direct action of nicotine in PPTg may be through GABAergic mechanisms that have been shown to influence nicotine self-administration preferentially compared to cocaine. OBJECTIVE: The purpose of these experiments was to examine several pharmacological manipulations that alter neuronal activity in the PPTg for their specificity or generality in nicotine versus cocaine reinforcement. METHODS AND RESULTS: Rats trained to self-administer nicotine or cocaine intravenously were prepared with brain microcannulae directed to the PPTg. Intra-PPTg microinfusions of the muscarinic agonist carbachol (0.1-1.0 microg), the micro opioid agonist DAMGO (0.005 and 0.05 microg), tetrodotoxin (5 ng) and neostigmine (0.5 nmol) each reduced the self-administration of nicotine and cocaine maintained on an FR5 schedule of reinforcement. The muscarinic antagonist scopolamine (0.1-1.0 microg) and the opioid antagonist CTOP (1 microg) did not affect self-administration, but reversed the effects of the respective agonist when co-administered with it. Carbachol and DAMGO were also tested in self-administration maintained on a progressive-ratio schedule; each agonist again reduced both nicotine and cocaine self-administration. CONCLUSIONS: PPTg manipulations are able to alter established self-administration of nicotine, which acts at the level of the ventral tegmental area and the PPTg itself, and cocaine, which acts through the mesolimbic dopamine system. These data suggest that the PPTg is an important substrate in drug dependence. PMID- 11875639 TI - Effects of intraperitoneal injections of saline on the alcohol and sucrose consumption of C57/BL10 mice. AB - RATIONALE: To determine the effects of multiple saline injections on alcohol drinking by male and female C57/BL10 mice with low preference for alcohol. OBJECTIVE: An investigation of the effects of multiple saline injections on alcohol consumption, with a comparison of corresponding effects on sucrose consumption. METHODS: The effects of a range of injection schedules on preference for 8% alcohol, or 1% sucrose, compared with tap water, were measured in two bottle choice tests. RESULTS: The multiple saline injection schedule significantly increased the alcohol preference, even when no alcohol was available during the injection period. The actual administration of fluid was not necessary for the increase in alcohol preference, since sham injections without fluid administration also increased alcohol preference. A single injection of saline did not alter the alcohol preference 3 weeks later. Daily saline injections for 3 weeks did not alter the consumption of the dilute sucrose solution. In the population of mice used, the preference for sucrose over water was found to follow a biphasic distribution, similar to that reported earlier in these mice for alcohol preference, but there was no correlation between alcohol preference and sucrose preference. CONCLUSIONS: The results suggest that lasting changes in the areas of the brain that specifically control alcohol intake are produced by repetition of a routine laboratory procedure. PMID- 11875640 TI - Modulation of the effects of alcohol on driving-related psychomotor skills by chronic exposure to cannabis. AB - RATIONALE: Many previous studies have reported that alcohol and cannabis produce additive psychomotor effects in acute combination, but few have explicitly tested whether chronic exposure to cannabis, in the absence of acute administration, alters the effects of alcohol on psychomotor performance. OBJECTIVES: To test whether long-term cannabis use modulates the effects of alcohol on psychomotor skills and self-reported mood and sensation. METHODS: Regular cannabis users (minimum: daily use for at least 3 years) and infrequent users (maximum: once monthly use for at most 3 years) were matched for sex, age, alcohol intake and other drug use (14 participants in each group). Participants received alcohol (females 0.35 g/kg; males 0.45 g/kg) and placebo drinks. By urinalysis, only regular users tested positive for metabolites of Delta(9)-tetrahydrocannabinol; breath alcohol levels were similar between groups. Participants were tested on a computerised tracking task that has been used to screen drugs for adverse effects on driving. The task involved tracking a moving target on a computer screen while simultaneously responding to occasional presentations of stimuli in the periphery of the screen. RESULTS: Tracking accuracy was similar for both groups after placebo, but alcohol caused a significant deterioration in performance among infrequent cannabis users relative to regular users. These changes were mirrored by significant changes in self-reported scores for dizziness, measured by visual analogue scales. Alcohol slowed reaction times, but not differentially between groups. CONCLUSIONS: For psychomotor skills relevant to driving, chronic cannabis use (in the absence of acute administration) does not potentiate the effects of alcohol. In fact, the superior tracking accuracy of regular users relative to infrequent users after alcohol, and their lower scores for dizziness, suggest that chronic cannabis use may instead confer cross-tolerance to specific effects of alcohol on behaviour. PMID- 11875641 TI - Co-release of noradrenaline and dopamine in the prefrontal cortex after acute morphine and during morphine withdrawal. AB - RATIONALE: Acute morphine and abstinence from chronic morphine have been shown to increase and to decrease extracellular dopamine (DA) in the nucleus accumbens, respectively. In contrast, extracellular DA in the prefrontal cortex (PFC) is not modified by acute morphine and is markedly increased during abstinence syndrome. OBJECTIVES: We investigated whether the peculiar behaviour of PFC DA might depend on the fact that extracellular DA originates not only from DA but, mainly, noradrenaline (NA) terminals. Accordingly, we studied if the effect of acute morphine and morphine-abstinence was modified by the inhibition of DA or NA neurons. METHODS: Extracellular DA and noradrenaline (NA) concentrations were determined by microdialysis in the PFC (densely innervated by DA) and in the parietal cortex (lacking DA afferents) both after acute morphine and in morphine dependent rats during naloxone-precipitated abstinence syndrome. Dialysate catecholamine levels were evaluated by high performance liquid chromatography (HPLC) with electrochemical detection. RESULTS: Acute morphine (5 mg/kg IP) reduced extracellular NA (by 30%) and failed to modify extracellular DA level in the PFC, but reduced both amines by 40% in the parietal cortex. The co administration of morphine and the D(2) agonist quinpirole (0.5 mg/kg IP) decreased both extracellular DA and NA by 40% in the PFC. In morphine dependent rats the administration of naloxone (1.0 mg/kg, SC) precipitated a typical abstinence syndrome associated with a concomitant dramatic increase in extracellular DA and NA by about 200 and 100%, respectively, in the PFC. The alpha(2)-adrenoceptor agonist clonidine (0.15 mg/kg IP) suppressed naloxone precipitated abstinence symptoms and brought both NA and DA output in the PFC to <50% baseline values. In contrast, quinpirole was totally ineffective. CONCLUSIONS: The results suggest that: a) morphine-stimulated DA release from DA terminals is compensated by reduced DA release from NA terminals; b) morphine abstinence-induced inhibition of DA release from DA terminals is overshadowed by a marked increase in DA released from NA terminals. Thus, the paradoxical response of PFC DA to morphine and morphine abstinence may be explained by the fact that extracellular DA in the PFC mainly represents the amine co-released from NA terminals. PMID- 11875642 TI - Fibroblasts enhance the invasive capacity of melanoma cells in vitro. AB - In previous experiments we have shown an enhanced expression of matrix metalloproteinase-1 (MMP-1) in fibroblasts obtained from the border of invasive melanoma in comparison to fibroblasts more distant from the tumour. In the study reported here we sought to determine whether melanoma-derived soluble factors are responsible for the stimulation of MMP-1 expression in fibroblasts. By real-time PCR and enzyme-linked immunosorbent assays, we demonstrated that the stimulation of fibroblasts with melanoma cell conditioned medium led to an increased expression of MMP-1 mRNA as well as MMP-1 protein, whereas melanoma cells themselves did not produce detectable amounts of MMP-1 protein. Basic fibroblast growth factor (bFGF) was detected as an important factor responsible for the enhanced expression of MMP-1 by fibroblasts after stimulation with melanoma cell conditioned medium. In a three-dimensional in vitro invasion assay, we demonstrated that fibroblasts are essential for melanoma cell invasion into a collagen I matrix. These findings support the hypothesis that stromal fibroblasts assist the invasion of melanoma cells through the extracellular matrix by producing elevated amounts of proteolytic enzymes after interaction with soluble factors (e.g. bFGF). PMID- 11875643 TI - Specific inhibition of human skin fibroblast chemotaxis to platelet-derived growth factor A-chain homodimer by transforming growth factor-beta1. AB - Platelet-derived growth factor (PDGF) and transforming growth factor-beta (TGF beta) have been suggested to play important roles in wound healing. We investigated the effect of TGF-beta1 on the mitogenic and chemotactic activities of PDGF A-chain homodimer (PDGF-AA) and B-chain homodimer (PDGF-BB) in primary cultures of human skin fibroblasts. TGF-beta1 inhibited the growth-promoting activity of both PDGFs. Proliferative responses to basic fibroblast growth factor and epidermal growth factor were also restricted by TGF-beta1. A Boyden chamber chemotaxis assay revealed that the chemotactic migration to PDGF-AA was inhibited by TGF-beta1 pretreatment, but in contrast, the response to PDGF-BB was not affected by the same treatment. Western blot analysis showed that TGF-beta1 downregulated PDGF alpha-receptors, but not beta-receptors, indicating that the isoform-specific inhibition of chemotaxis is related to differential effects of TGF-beta1 on PDGF receptor expression. The present findings suggest that TGF beta1 may act antagonistically towards PDGFs in humans under certain conditions, and this antagonistic nature of TGF-beta1 must be considered when it is applied to human wounds as a therapeutic agent. PMID- 11875645 TI - Effect of vitamin D3 on the increased expression of Bcl-xL in psoriasis. AB - Psoriasis is a chronic skin disease characterized by epidermal hyperproliferation, which may be regulated by several mechanisms including apoptosis. In this study, we detected DNA fragmentation by the terminal deoxynucleotide transferase-mediated dUTP nick-end labeling (TUNEL) method and immunohistochemically examined the expression of Bcl-x and Bax in psoriasis. We determined the expression of bcl-xL mRNA by RT-PCR, and also determined the effect of vitamin D(3) (VD3) on bcl-xL mRNA expression in cultured normal human keratinocytes by RT-PCR, and the expression of Bcl-xL in psoriatic lesions before and after topical application of VD3. A large number of TUNEL-positive cells as well as Bcl-xL - and Bax-positive cells were observed throughout the epidermis in psoriatic lesions. Whereas, in nonlesional and normal skin, only a few TUNEL positive cells were observed and only the lower epidermis showed positive staining for Bcl-x and Bax. We also observed higher expression of bcl-xL mRNA in psoriatic lesions than in nonlesional and normal skin. The expression of bcl-xL mRNA in cultured normal human keratinocytes stimulated or not with IFN-gamma and PMA was suppressed by VD3 in a dose-dependent manner, and the expression of Bcl xL, but not Bax, in psoriatic lesional skin decreased after topical application of VD3 for 4 weeks. In conclusion, it is suggested that the apoptotic process in psoriatic lesions is in part regulated by Bcl-xL, and decreasing the expression of Bcl-xL by treatment with VD3 might ameliorate psoriatic lesions by contributing to the completion of the apoptotic process. PMID- 11875644 TI - Increased nerve growth factor- and tyrosine kinase A-like immunoreactivities in prurigo nodularis skin -- an exploration of the cause of neurohyperplasia. AB - Neurotrophins and their receptors play an important role in cutaneous nerve development and reconstruction after injury. Recent developments indicate that this group of molecules not only exert a neurotrophic action, but are also involved in immune responses and inflammation. Prurigo nodularis is a skin disease characterized by neurohyperplasia and intense itch. In the present study, the localization and distribution of nerve growth factor (NGF) and its receptors were explored by immunohistochemical methods, with the aim of detecting the cause of the neurohyperplasia in the disease. In normal healthy volunteers and in uninvolved skin, NGF immunoreactivity was seldom seen in the basal layer of the epidermis or in the dermis. In prurigo nodularis skin, there was also very little NGF immunoreactivity in the epidermis. However, in the dermis, a huge number of cells showed an NGF-like immunoreactivity. In normal skin of healthy volunteers, only a weak staining for tyrosine kinase A (trkA) was seen in the epidermis, whereas in the dermis, there was no trkA staining seen at all. However, in the prurigo nodularis tissue, the hyperplastic nerves clearly showed trkA immunoreactivity, and it seemed that the staining was only present in the axons. By NGF and p75 NGF receptor double-labelling, both immunoreactivities showed weak staining in the epidermis and dermis of normal skin. However, in the dermis of prurigo nodularis, strong staining for both NGF and NGF receptor antibodies was seen. NGF receptor-immunoreactive nerves were more dense in areas where there were more NGF-immunoreactive cells. The results indicate that in prurigo nodularis skin, NGF is overexpressed, locally infiltrated inflammatory cells may be the source of this NGF, and NGF and its receptors may contribute to the neurohyperplasia of the disease. PMID- 11875646 TI - Multiple biological effects of inhibitors of arachidonic acid metabolism on human keratinocytes. AB - BACKGROUND: Various compounds that inhibit processing of arachidonic acid (AA) are being intensively tested for their antitumour activity. However, the mechanisms responsible for such activity remain rather elusive. To approach this issue, we examined the effects of several structurally different inhibitors of AA metabolism in the human keratinocyte HaCaT cell line. METHODS: Several parameters were determined in HaCaT cells exposed to increasing concentrations of the inhibitors for 24 and/or 48 h. These included (1) oxidoreductase activity, total protein mass and cell cycle distribution to assess cell proliferation, (2) degradation of PARP protein to assess apoptosis, and (3) cell morphology, distribution of F-actin and expression of cytokeratins and E-cadherin to evaluate changes in differentiation status. RESULTS: While eicosatetraynoic acid (ETYA), nordihydroguaiaretic acid (NDGA), esculetin and MK-886 reduced proliferation of HaCaT cells, the cyclooxygenase inhibitors indomethacin and piroxicam had no such effects. Esculetin and NDGA arrested cells in S phase, and ETYA and MK-886 delayed cell progression through G(1) phase. Higher concentrations of NDGA, MK886 and/or ETYA caused cleavage of PARP. No changes in the expression of cytokeratins and E-cadherin were observed upon treatment with any of the inhibitors. However, esculetin induced redistribution of F-actin accompanied by increased cell adhesion and size. CONCLUSION: Our findings indicate that, in addition to their ability to inhibit cell proliferation and to induce apoptosis, lipoxygenase inhibitors and/or ETYA may also elicit other important physiological responses in HaCaT keratinocytes. PMID- 11875647 TI - Ultraviolet B irradiation increases keratin 5 and keratin 14 expression through epidermal growth factor receptor of SV40-transformed human keratinocytes. AB - Keratin intermediate filaments are heteropolymers composed of type I and type II keratins. Ultraviolet B (UVB) irradiation induces keratin expression by keratinocytes. Using SV40-transformed human keratinocytes (SVHK), we investigated the effect of UVB irradiation on keratin expression. UVB irradiation (10 mJ/cm(2)) increased keratin 5 and keratin 14 mRNAs and proteins without affecting cell viability. Upregulation of keratin 5 and keratin 14 was dependent on the dose of radiation: the effect was observed at 5 mJ/cm(2) and the maximal effect was observed at 10 mJ/cm(2). Higher UVB doses (more than 10 mJ/cm(2)) were cytotoxic. Expression of keratin 1 and keratin 10 was marginal in SVHK and was not affected at either the mRNA or protein level by UVB. The stimulatory effects on keratin 5 and keratin 14 expression were also observed in cultured normal human keratinocytes (NHK) and HaCaT keratinocytes. The tyrosine kinase inhibitor, genistein, and the epidermal growth factor (EGF) receptor inhibitor, AG1429, significantly suppressed the increase in expression of keratin 5 and keratin 14 by SVHK. In contrast, the suppressive effect was not observed with the protein kinase C inhibitor, H-7. Furthermore, pretreatment with neutralizing anti-EGF receptor antibody also suppressed UVB-induced keratin 5 and keratin 14 expression by SVHK, NHK and HaCaT cells. UVB irradiation did not affect the steady-state expression of TGF-alpha by SVHK. Immunoprecipitation and immunohistochemical studies revealed that UVB irradiation induced EGF receptor activation in the absence of EGF and TGF-alpha. These results indicate that UVB increases keratin 5 and keratin 14 expression through direct activation of the EGF receptor in SVHK. PMID- 11875648 TI - The effect of extracorporeal photochemotherapy on alpha1(I) and alpha1(III) procollagen mRNA expression in systemic sclerosis skin tissue. PMID- 11875650 TI - Meeting Calender 293/12. PMID- 11875649 TI - Ultrastructural localization of NF1 tumor suppressor protein in human skin. PMID- 11875651 TI - Transient activation of the micro1 homeobox gene family in the sea urchin ( Hemicentrotus pulcherrimus) micromere. AB - The animal-vegetal (A-V) axis of sea urchin embryos is morphologically evident at the 16-cell stage of development. Mesomeres, macromeres, and micromeres are arrayed along the A-V axis. The vegetal micromere differentiates into the skeletogenic mesenchyme and functions as a signaling center. To date, no zygotic or maternally specified gene with restricted expression in the micromere at the 16-cell stage has been reported. We performed subtraction PCR and dot blot hybridization using poly(A)+ RNA extracted from the micromere (tester) and the mesomere (driver) in order to identify micromere-specific genes. Using a cDNA fragment identified in this screen, we isolated four similar but distinct cDNA clones from a library, which corresponded to a group of genes that we refer to as the micro1 family. The micro1 family encoded putative transcription factors with a homeodomain which had 87-95% identity between family members. The most highly conserved protein was encoded by PlHbox12 from Paracentrotus lividus (71-76% identity among family members). Northern blot hybridization and in situ hybridization demonstrated that micro1 was transiently activated during the early cleavage stages and that the transcript was restricted to the micromere. Thus, the expression domain was complementary to that of PlHbox12 along the A-V axis. The micro1 gene family has at least six loci, including polymorphic alleles, which are probably clustered in the genome. PlHbox12 and micro1 constitute a novel family of paired-like class homeobox genes. Phylogenetic analyses suggest that PlHbox12/micro1 evolved exceptionally rapidly. PMID- 11875652 TI - Primordial germ cells originate from the endodermal strand cells in the ascidian Ciona intestinalis. AB - The origin of germ cells in the ascidian is still unknown. Previously, we cloned a vasa homologue (CiVH) of Ciona intestinalis from the cDNA library of ovarian tissue by polymerase chain reaction and showed that its expression was specific to germ cells in adult and juvenile gonads. In the present study, we prepared a monoclonal antibody against CiVH protein and traced the staining for this antibody from the middle tailbud stage to young adulthood. Results showed that positive cells are present in the endodermal strand in middle tailbud embryos and larvae. When the larval tail was absorbed into the trunk during metamorphosis, the CiVH-positive cells migrated from the debris of the tail into the developing gonad rudiment, and appeared to give rise to a primordial germ cell (PGC) in the young juvenile. The testis rudiment separated from the gonad rudiment, the remainder of which differentiated into the ovary. PGCs of the testis rudiment and the ovary rudiment differentiated into spermatogenic and oogenic cells, respectively. When the larval tail containing the antibody-positive cells was removed, the juveniles did not contain any CiVH-positive cells after metamorphosis, indicating that the PGCs in the juvenile originated from part of the larval tail. However, even in such juveniles, positive cells newly appeared in the gonad rudiment at a later stage. This observation suggests that a compensatory mechanism regulates germline formation in C. intestinalis. PMID- 11875653 TI - Heterospecific transgenesis in Drosophila suggests that engrailed.a is regulated by POU proteins in the crustacean Sacculina carcini. AB - Almost all knowledge of the regulation of segmentation genes in arthropods comes from Drosophila. In order to study the regulation of the segment-polarity gene engrailed in a non-insect arthropod we focussed on putative regulatory regions of the engrailed.a (en.a) gene in the barnacle crustacean Sacculina carcini. In this animal, en.ais expressed in segmental stripes like the engrailed genes of other arthropods. As transgenesis in Sacculina is not possible at present, we have used Drosophila as a test tube. The Sacculina en.aintron is able to induce a specific expression of lacZin the Drosophila wing imaginal disc.This pattern is not an engrailed-like pattern, but does suggest that some Drosophila transcription factors interact with the Sacculina en.a intron. We show that two DrosophilaPOU proteins, Nubbin and VVL, and Engrailed itself bind to the Sacculina en.a intron in vitro and that they regulate this expression in vivo. The conservation of POU protein binding sites in metazoans suggests that Sacculina POU proteins could recognize the same sequences. Hence, we looked at the expression of nubbin and vvlhomologues in Sacculinalarvae. Indeed, their expression patterns are consistent with a putative regulatory function on en.a in segments and appendages. Remarkably, the vvl homologue is expressed in Sacculina in a striking striped pattern that is very different from the vvl pattern in Drosophila embryos, and is complementary to the Sacculina en.a pattern. These experiments suggest that the Sacculina engrailed.a gene is regulated by POU proteins. PMID- 11875655 TI - Fate map of the eye-antennal imaginal disc in the stalk-eyed fly Cyrtodiopsis dalmanni. AB - Hypercephaly, in the form of lateral extensions of the head capsule, is observed in several families of Diptera. A particularly extreme form is found in diopsid stalk-eyed flies, in which both eyes and antennae are laterally displaced at the end of eyestalks. We have studied the developmental basis of this exaggerated morphology in Cyrtodiopsis dalmanni. Diopsid eye-antennal imaginal discs are divided into anterior and posterior portions, which are joined by a narrow "disc stalk" of intervening tissue. We established a fate map for this disc by cutting it into fragments and culturing them in vivo by injecting them into host larvae. The adult eye and dorsal head capsule structures, including the eyestalk and the ocelli, are derived from the posterior portion of the disc, while ventral adult structures such as the antenna and the palpus are derived from the anterior portion of the disc. Thus both posterior and anterior disc portions give rise to structures that are widely separated in the adult head. Moreover, structures that are adjacent in the adult are derived from different regions of the disc. These results confirm and extend previous conclusions about regional identity in diopsid eye-antennal discs that were based on the analysis of molecular markers. PMID- 11875654 TI - Isolation of cDNA clones for mRNAs transcribed zygotically during cleavage in the ascidian, Halocynthia roretzi. AB - The ascidian larva consists of a relatively small number of different cell types, and the cell lineages during embryogenesis have been well described. The clonal restriction of developmental fate takes place considerably early in development. The fates of most of the blastomeres become tissue-restricted by the 110-cell stage, just before the onset of gastrulation. To elucidate the molecular basis of the early events of fate determination in the ascidian Halocynthia roretzi, we isolated the genes for which zygotic expression is initiated during the early cleavage stages. Here we report 18 genes isolated by subtractive hybridization screening between 110-cell embryos and fertilized eggs. The expression of most (13) of the genes was initiated at the 32-cell stage. The genes were subdivided into three groups according to their spatial expression patterns. The first group included clones expressed throughout almost the entire embryo. The second and third groups represented clones expressed mainly in the animal hemisphere and in a subset of vegetal blastomeres, respectively. One of the genes, HrHesl1, encoded a polypeptide containing the bHLH domain that is similar to those of the Hairy/Enhancer of split/Deadpan family of transcriptional repressors. HrHesl1 was expressed exclusively in epidermal precursor cells during cleavage. Another gene named HrWnt-5 beta was expressed in muscle precursors. PMID- 11875656 TI - mPet-1, a mouse ETS-domain transcription factor, is expressed in central serotonergic neurons. AB - Here we describe the expression pattern of a previously unknown mouse gene mPet 1. The isolated cDNA codes for an ETS-domain transcription factor of 237 amino acids in length, which is localized to the nucleus. mPet-1 is a member of the winged helix transcription factor gene family like its rat homologue Pet-1 and the human homologue FEV. The start ATG of mPet-1 and the size of the predicted protein are identical to the human FEV. The mPet-1 protein is clearly smaller since it lacks the first 103 N-terminal amino acids of rat Pet-1. mPet- 1 is expressed in central serotonergic (5-hydroxytryptaminergic) neurons located in the mes-/metencephalic raphe nuclei from E11 on until adulthood. In these regions mPet-1 expression co-localizes precisely with the serotonin transporter (Sert),which it initially precedes. Interestingly, mPet-1 was not found in neurons transiently expressing Sert. PMID- 11875657 TI - Developmental expression of cardiac myosin-binding protein C in Xenopus. AB - We have isolated the Xenopus homologue of cardiac myosin-binding protein C ( cMyBP-C) and describe its expression during early embryogenesis. cMyBP-C is expressed in both somites and heart at the time these tissues differentiate. Expression in the somites declines in older tadpoles. There is a high degree of conservation in residues where mutations causing hypertrophic cardiomyopathy have been identified in humans. PMID- 11875658 TI - Developmental expression of ascidian neurotransmitter synthesis genes. I. Choline acetyltransferase and acetylcholine transporter genes. AB - To identify cholinergic neurons, we isolated a choline acetyltransferase (Ci ChAT) gene from Ciona intestinalis by PCR methods. In the cloning process, we also obtained the gene encoding the vesicular acetylcholine transporter (Ci vAChTP). These two genes shared the same 5'-UTR sequence as well as similar expression patterns. In both cases, the gene expression was first detected by whole-mount in situ hybridization in the anterior-dorsal region of the caudal nerve cord at the early tailbud stage. In the larva, the expression was seen in several cells of the visceral ganglion. These results suggest that ascidian larval motor neurons exist in the visceral ganglion. PMID- 11875659 TI - Spatial and temporal pattern of Fgf-8 expression during chicken development. AB - This study analyzes the temporal and spatial expression pattern of Fgf-8 over a continuous series of developmental stages. Special emphasis is laid on the paraxial mesoderm where Fgf-8 expression is highly dynamic. Whereas the anterior portion of the unsegmented mesoderm is devoid of expression, Fgf-8 is upregulated in the posterior half of a newly formed somite. Soon after somite formation, this highly localized expression gives way to a more diffuse pattern of Fgf-8 expression at low levels in presumptive sclerotomal cells. During later somite maturation, transcripts become restricted to the myotome. Co-staining with the myotome marker MyoD reveals that Fgf-8 expression defines a subpopulation of muscle precursor cells. PMID- 11875660 TI - Sequential observations of exencephaly and subsequent morphological changes by mouse exo utero development system: analysis of the mechanism of transformation from exencephaly to anencephaly. AB - Anencephaly has been suggested to develop from exencephaly; however, there is little direct experimental evidence to support this, and the mechanism of transformation remains unclear. We examined this theory using the exo utero development system that allows direct and sequential observations of mid- to late gestation mouse embryos. We observed the exencephaly induced by 5-azacytidine at embryonic day 13.5 (E13.5), let the embryos develop exo utero until E18.5, and re observed the same embryos at E18.5. We confirmed several cases of transformation from exencephaly to anencephaly. However, in many cases, the exencephalic brain tissue was preserved with more or less reduction during this period. To analyze the transformation patterns, we classified the exencephaly by size and shape of the exencephalic tissue into several types at E13.5 and E18.5. It was found that the transformation of exencephalic tissue was not simply size-dependent, and all cases of anencephaly at E18.5 resulted from embryos with a large amount of exencephalic tissue at E13.5. Microscopic observation showed the configuration of exencephaly at E13.5, frequent hemorrhaging and detachment of the neural plate from surface ectoderm in the exencephalic head at E15.5, and multiple modes of reduction in the exencephalic tissue at E18.5. From observations of the vasculature, altered distribution patterns of vessels were identified in the exencephalic head. These findings suggest that overgrowth of the exencephalic neural tissue causes the altered distribution patterns of vessels, subsequent peripheral circulatory failure and/or hemorrhaging in various parts of the exencephalic head, leading to the multiple modes of tissue reduction during transformation from exencephaly to anencephaly. PMID- 11875661 TI - Vasculogenesis and angiogenesis in the subcardinal venous plexus of quail mesonephros: spatial and temporal morphological analysis. AB - Vasculogenesis and angiogenesis are involved in a coordinated program for the development of the mesonephric subcardinal venous plexus of quail embryo. Vasculogenesis occurs between days 3 and 4 of incubation, while angiogenesis takes place from day 5 to day 7. Examination of vascular corrosion casts and whole mounts, and tissue sections labelled with specific markers to hemangioblast lineage (QH1, LEP100 and AcPase activity), allowed us to distinguish six phases in the formation of subcardinal plexus. (1) Appearance of isolated angioblast like cells where the subcardinal plexus will form. (2) Alignment of angioblast like cells into cellular strands. (3) Formation of compact vascular cords by association of angioblast-like strands. (4) Polygonal interconnection of vascular cords to constitute the primary subcardinal plexus. In this stage, isolated angioblast-like cells were present inside inter-vascular spaces. (5) The splitting of primary inter-vascular spaces by angiogenic sprouts to form secondary subcardinal plexus (outward angiogenesis). Isolated angioblast-like cells were not present in this stage. (6) Expansion of the secondary subcardinal plexus by insertion of slender transcapillary tissue pillars (inward angiogenesis) and angiogenic sprouts. We also describe three morphogenetic gradients during the development of the subcardinal plexus: ventral-to-dorsal, cranial-to-caudal and lateral-to-medial. PMID- 11875663 TI - Diaphorase-positive neurons in the cingulate cortex of human fetuses during the second half of gestation. AB - In this work, the time course of appearance, distribution and morphology of diaphorase-positive neurons were studied in the developing cingulate cortex of the human brain during the second half of gestation. Five human fetuses at 18, 20, 25, 30 and 35 weeks postovulatory (wpo) were examined. The brain tissue was reacted by an indirect histochemistry protocol for detection of NADPH-diaphorase activity. Labeled neurons were identified at the microscope and documented photographically or by computer-aided charts. We have found that heavily labeled neurons (type I) first appear in the subplate (SP) between 20 and 25 wpo, and in the cortical plate (CP) between 25 and 35 wpo. By 35 wpo, CP neurons were both type I and type II (lightly labeled neurons). In addition, we observed 4 different morphological types among subplate neurons, very similar to callosally projecting subplate cells (as described previously by our group). We concluded that medial nitridergic neurons of humans appear prenatally according to the usual gradient of cortical maturation -- first in the subplate and later in the cortical plate. Also, we suggest that some of the diaphorase-positive neurons in the transient subplate could possibly be callosal. PMID- 11875664 TI - Expression of Kin, a nuclear protein binding to curved DNA, in the brain of the frog (Rana esculenta), turtle (Trachemys scripta), quail (Coturnix coturnix) and mouse (Mus musculus). AB - The distribution of Kin protein, the vertebrate homologue of the bacterial recA nuclear protein involved in illegitimate recombinant DNA repair and gene regulation, was analysed in the brain of the mouse, quail, turtle and frog by immunocytochemical methods. The protein was expressed in all brains, but not in a uniform manner. Immunoreactivity was absent from major fibre tracts. In the cerebral nuclei, immunolabelling in the various species showed an important variation. A comparative analysis, based on the homologies between different brain structures in these species, showed that this variation was not due to interspecific variation but that of an ancestral pattern of distribution of Kin protein. It is also shown that whatever the species examined, Kin protein is consistently more highly expressed in those regions of the brain with a conservative evolutionary history (e.g. the olfactory and limbic systems, the hypothalamus, the monoaminergic system, the cerebellum, and the nuclei of sensory and motor cranial nerves). The protein is markedly less heavily expressed in the dorsal striatum and the sensory nuclei of the thalamus. PMID- 11875666 TI - Occlusion and subsequent re-canalization in early duodenal development of human embryos: integrated organogenesis and histogenesis through a possible epithelial mesenchymal interaction. AB - Histogenesis of the duodenum, especially changes in the epithelium in relation to temporal occlusion and re-canalization of the lumen, was investigated by light microscopy together with morphometric analysis, as well as by scanning and transmission electron microscopy of 133 externally normal human embryos ranging from Carnegie stage 12 to 23. A series of morphogenetic events passed the duodenum in a cranio-caudal (proximo-distal) wave like fashion during the period examined. They included: (1) a decrease in the caliber and area of the lumen, (2) 'occlusion' of the lumen, (3) vacuole formation, (4) 're-canalization' and villi formation. The only exemption to this rule was that, in the upper part of the duodenum, the lumen was not obliterated in the embryos examined. Morphometric analyses revealed that both the area of the epithelium and the number of epithelial cells decreased during the 'occlusion' phase. This result suggests that, unlike the classical view, epithelial cell proliferation does not play an important role in occluding the lumen, but the predominant morphogenetic event during this phase is convergence of the epithelial cells to elongate the duodenum. Apoptosis, contrary to some classical views, decreased during the 're canalization' phase, and it appeared to be involved in the formation of the small lumens in the epithelial 'plug' and in villi formation, but not in enlarging the secondary lumens. The secondary small lumens in the occluded lumen were frequently formed near the border between the central 'plug' and peripheral basal cells on the basement membrane. This and other findings of concentric differentiation in both the epithelial and mesenchymal layers suggested a possible control mechanism by the epithelium-mesenchymal interaction on human duodenal morphogenesis and histogenesis. The present electron microscopic observations also provided details on the mechanisms involved in the enlargement of the secondary lumen and differentiation of villi. The implications of these findings to duodenal anomalies are also discussed. PMID- 11875667 TI - The genus Acipenser as a model system for vertebrate urogenital development: nephrostomial tubules and their significance for the origin of the gonad. AB - Early gonadal development was studied in the sterlet, Acipenser ruthenus, by means of histological and semithin serial sections and scanning electron microscopy. Special attention was given to the role of opisthonephric nephrostomial tubules and their coelomic funnels (nephrostomes, coelomostomes) in the origin of the gonad. Specimens of about 1 mm in length (about 7 days post hatching) have a continuous kidney complex (holonephros) that extends from the branchial region to the level of the cloaca and may be divided into a cranial pronephros and a caudally following opisthonephros, with no overlapping of either portion. In specimens of 10 to 25 mm in length the regression of pronephros and cranial opisthonephros can already been seen; as a consequence, these parts of the kidney complex are not involved in gonadogenesis. The initial gonadal anlage is seen in specimens of 30-40 mm in length. The somatic cells of the gonadal primordium develop from the medial lips of segmentally arranged opisthonephric nephrostomes situated in a line that extends from the level of the stomach through that of the spiral valve. The nephrostomes involved in this process belong to the first-order set of nephrons, since nephrons of higher order that arise continuously from blastema cells during further growth of the animals never send nephrostomial tubules to the coelomic surface. The cells of the medial nephrostomial lips proliferate by many mitoses. They grow over and surround the large germ cells that have accumulated on the medial side of the nephrostomes. The proliferating nephrostomial cells are elongated in shape, and their long axes are oriented in the cranio-caudal direction. By their size, shape and arrangement they replace not only the flat, polygonal, mesothelial cells of lateral plate origin on the medial side of the nephrostomes, but also those in the interstices in between. The result is the formation of a continuous gonadal crest situated medially from the nephrostomial line. At 100-130 mm in length, the gonadal crest has reached the stage of a gonadal fold that is attached to the dorsal body wall by a thin mesogonadium. Stroma cells and blood vessels start to invade the gonadal fold. At 240-290 mm in length, parts of the gonadal fold are converted into a fat body, a structure that is typically present in adult sturgeons of either sex. Nephrostomial tubules and their nephrostomes are visible only for a short period in Acipenser ruthenus. They are completely developed in specimens of 25-40 mm in length and start continuous retrogression immediately afterwards. Therefore, they cannot play an important role in excretion. Their only purpose apparently is to function as precursor tissue for other organs, such as the gonad, in establishing a route by which cells of the intermediate mesoderm can gain access to and spread out over the coelomic surface. In conclusion, the observations made in Acipenser prove that this species is a most suitable model to explain the origin of the gonad from opistho/mesonephric nephrostomial tubules in vertebrates with a less-developed or early regressing pronephros. Most mammals, including man, belong to this category. PMID- 11875668 TI - Posttransplant lymphoproliferative disease in a child: clinical and molecular characterization. AB - We report a 12-year-old girl suffering from end-stage renal disease due to focal segmental sclerosis and retardation of statomotoric and mental development of unknown origin. Renal transplantation (TX) was performed 7 months after initiation of peritoneal dialysis at the age of 11 years. Immunosuppressive therapy included cyclosporine A, mycophenolate mofetil and methylprednisolone. The patient developed spiking fever up to 40 degrees C without signs of infection 10 months after TX. Kidney function remained stable but ultrasound examination and CT-scan showed hypodense masses within both liver and spleen. Epstein-Barr virus (EBV) polymerase chain reaction (PCR) results with a high number of copies (20 x 10(6) copies/ml blood) against the background of a previous EBV infection (IgG positive, IgM negative) made the diagnosis of EBV-reactivation likely. Splenectomy was performed. Examination of the spleen showed EBV-associated polymorphic posttransplant lymphoproliferative disease (PTLD) with predominant B cell proliferation and monoclonal VH3-rearrangement of the IgG heavy chain locus. Therapy with acyclovir was introduced and immunosuppression was reduced. No rejection episode occurred. Body temperature normalized and the patient recovered over a 3-month period. EBV-PCR in plasma was negative (0.02 x 10(6) copies/ml blood) 12 weeks after reduction of immunosuppression. The liver masses completely resolved after 27 months. After a total follow-up of 36 months the child remains in good health. PMID- 11875669 TI - Humoral immunity and frequency of peritonitis in chronic peritoneal dialysis patients. AB - This study was designed to investigate the humoral immune status and immunological abnormalities in relation to the pathogenesis of peritonitis in patients undergoing continuous ambulatory peritoneal dialysis (CAPD). The levels of serum and dialysate total protein, immunoglobulins (IgG, IgA, IgM, and IgG subgroups), and C3 were measured at baseline and at the end of the study in 12 children and 10 adult patients on CAPD. The relationship between the levels of immunoglobulin and C3 and the frequency of peritonitis was investigated. Peritonitis occurred at a frequency of one episode every 3.9 and 6.3 patient months in children at the beginning and at the end of the study, respectively; however, the respective rates for adult patients were one episode every 5.6 and 9.8 patient-months. The levels of serum immunoglobulin and C3 were normal in adults at the beginning and at the end of the study. However, the levels of serum IgG were low in 4 of 12 pediatric patients. Of the 4 patients with low total IgG, IgA was low in 1 and IgM was low in 2 patients. Of 4 patients with low total IgG, 2 also had low IgG1 and IgG2. However, 3 of the patients with normal IgG had low IgG2 levels at the end of the study. The levels of C3 were low in 6 of 12 and in 3 of 10 pediatric patients at the beginning and at the end of the study, respectively. The protein levels of dialysate were increased at 1-year follow-up in both groups, but the difference was only statistically significant in adult patients (P<0.05). No correlation was found between the levels of serum and dialysate immunoglobulin and the frequency of peritonitis in either group. In this study, hypogammaglobulinemia and hypocomplementemia were determined in pediatric patients undergoing CAPD. Although these immunological deficits are not major factors in the occurrence of CAPD-related peritonitis, they may contribute to the frequency of peritonitis. PMID- 11875670 TI - Renal failure after anti-D globulin treatment of idiopathic thrombocytopenic purpura. AB - Idiopathic thrombocytopenic purpura (ITP) is a disorder of rapid destruction of antibody-coated platelets. Anti-D immune globulin has been used for treatment of ITP in the United States since 1995. Initial studies identified no significant side effects of treatment. However, a recent report highlighted occasional episodes of intravascular hemolysis after anti-D immune globulin. We describe two children with ITP who developed acute renal failure (ARF) after treatment with anti-D immune globulin and also analyze ten additional cases of ARF reported to the manufacturer, Cangene Corporation, through postmarketing surveillance. All episodes of ARF were associated with intravascular hemolysis. Four patients required dialysis. Patient age ranged from 1 to 82 years, but those requiring dialysis were all under age 15 years. Several patients with ARF had preexisting creatinine elevation. Three of the patients with ARF had serologic evidence of acute Epstein-Barr virus (EBV) infection. Renal biopsy in one patient showed acute tubular necrosis, with findings consistent with pigment nephropathy. Anti-D immune globulin, used to treat ITP, may be associated with intravascular hemolysis and resultant ARF. Renal function should be monitored in patients with evidence of intravascular hemolysis. Children and adolescents may have increased risk of ARF requiring dialysis. PMID- 11875671 TI - Toxicity of chloroacetaldehyde is similar in adult and pediatric kidney tubules. AB - The nephrotoxicity of chloroacetaldehyde (CAA), one of the main products of hepatic ifosfamide metabolism, was compared in isolated human pediatric and adult renal tubules. Tubules metabolizing lactate were incubated in the presence of various concentrations of CAA (0.1-0.5 mM). Both at low, clinically relevant (0.2 mM), and at higher concentrations (0.3, 0.4 and 0.5 mM), CAA induced a cellular depletion of thiol compounds, i.e. glutathione, coenzyme A and acetyl-coenzyme A that are involved in CAA detoxication and cellular energy metabolism, respectively. The toxicity to renal cells was clearly observed in the presence of 0.4 and 0.5 mM CAA, which led to a fall of the cellular ATP level, to the accumulation of pyruvate and the inhibition of glucose synthesis from lactate. Inhibition of lactate uptake and an increase in the release of lactate dehydrogenase were observed only in the presence of 0.5 mM CAA. The sensitivity of pediatric tubules to the toxic effects of CAA and the rate of their CAA uptake were not statistically different from those found in adult tubules. It is concluded that an increased susceptibility of pediatric tubules to CAA toxicity cannot be put forward to explain the increased risk for ifosfamide-induced nephrotoxicity in children relative to adults. PMID- 11875672 TI - Impact of age on reference values for serum concentration of cystatin C in children. AB - Recently, cystatin C (cyst C) was proposed for the assessment of glomerular filtration rate, being more accurate than creatinine determination. Reference intervals for cyst C do not vary with age and sex, like creatinine values. Elevated values of cyst C were reported for pre- and full-term infants. Nevertheless, the age cut-off for stable cyst C values i.e., age independence, remains under discussion. Therefore, we conducted a prospective study in 98 healthy children, 51 being under 18 months of age. Cyst C was determined by the nephelometric method. The infants under the age of 18 months had a higher mean serum cyst C value (0.94 +/- 0.24 mg/l) than the older children (0.65 +/- 0.19 mg/l). There was a negative correlation between age and cyst C in the infant group under the age of 18 months (r(2)=0.631, P<0.01). Our results indicate that mean serum cyst C is higher in infants than older children; the age cut-off appears over the age of 1 year of life, presumably reflecting kidney maturation. Our study does not allow accurate assessment of the age cut-off at 18 months or 36 months. PMID- 11875673 TI - Quantitation of microalbuminuria using random urine samples. AB - Microalbuminuria is a harbinger of progressive renal disease and cardiovascular complications in patients with diabetes mellitus. The method most commonly used to measure microalbuminuria relies on a timed urine collection, either a 24-h or overnight specimen, which is time-consuming, cumbersome, and often inaccurate. We compared microalbumin-creatinine ratio (U(MA)/U(Cr)) in a random urine sample obtained after the first voided morning specimen with the quantity of microalbumin in a 24-h collection to determine whether the U(MA)/U(Cr) correlates with the microalbumin content of 24-h urine collection. In a study of 124 urine samples from 97 pediatric patients with type I diabetes, daily microalbumin excretion varied from 7 to 108 mg/24-h with a mean of 55.7 +/- 18.2 mg and U(MA)/U(Cr) ranged from 5 to 59 microg/mg with a mean of 39.4 +/- 11.3. An excellent correlation was found between the microalbumin excretion measured in 24 h urine collections and the random urine U(MA)/U(Cr) specimens (r=0.89, P<0.001). All patients who excreted more than 30 mg microalbumin in the 24-h specimen also had a U(MA)/U(Cr) of more than 20 microg/mg in the randomly voided sample. Microalbuminuria was unlikely if the U(MA)/U(Cr) was below 20 microg/mg. The results of this study indicate that the measurement of U(MA)/U(Cr) in a second voided morning specimen is a simple and reliable method for monitoring microalbuminuria in diabetic patients and may replace the need to assess quantitative microalbumin excretion on 24-h urine collections. PMID- 11875674 TI - Nutrition in children with preterminal chronic renal failure. Myth or important therapeutic aid? AB - Nutrition has been believed to be an important therapeutic instrument in children with chronic renal failure (i) for improving growth, and (ii) for slowing down the deterioration of renal function. The therapeutic strategies for both targets may be conflicting, at least in part, since a high calorie intake is needed for optimal growth, whereas a low protein diet, which was believed to protect renal function, places patients at risk of low calorie intake. Dietary manipulations for optimal growth are mainly effective in infants with chronic renal failure. However, growth remains suboptimal even with an energy intake above 80% of RDA. Although a low protein diet is able to slow down the rate of deterioration in renal function in rodent studies, the results of prospective clinical studies were disappointing at least for an observation period up to three years. The conclusions out of meta-analyses of these clinical studies in adults are contradictory. The progression rate was not significantly influenced by protein restriction, whereas renal replacement therapy could be postponed. However, the latter seems to be the effect of weakening uremic symptoms during the phase of end-stage renal failure. According to our present knowledge it is not justified to prescribe special diets to children early in the course of chronic renal failure, but the composition of their nutrition should follow the general concept of an optimal mixed diet. PMID- 11875675 TI - In utero exposure to immunosuppressive drugs: experimental and clinical studies. AB - Over the last few decades, the number of pregnant women under immunosuppressive (IS) therapy following transplantation or autoimmune diseases has increased. At first, IS drugs, including prednisone, azathioprine, and cyclosporine A were used, but now new molecules such as tacrolimus and mycophenolate mofetil have appeared. These IS drugs cross the placental barrier and enter into the fetal circulation, which poses a risk for fetal development. Experimental data have shown that IS drugs often have deleterious effects on fetuses, while human data have reported an increased rate of abortion, prematurity, intrauterine growth retardation (IUGR), and low birth weight, without significant increases in malformation rates. However, only limited information is available about the newly used molecules. Although fetal and neonatal data are reassuring, long-term effects of IS drugs on fertility, immune response and renal function, as well as the consequences of prematurity and IUGR, should be monitored. PMID- 11875676 TI - Clinical quiz. Peritoneal pseudocyst. PMID- 11875677 TI - International Perinatal Nephrology Symposium, 20-21 June 2001, Lausanne, Switzerland. AB - The 5th International Symposium on Prenatal Nephrology returned this year to Lausanne, Switzerland, after being held in the 2 previous years across the border in France (Strasbourg 1999 and Dijon 2000). The main topics presented for discussion to the approximately 80 participants (from 13 countries) dealt this year -- in English and in French -- with several aspects of acute renal failure (ARF) in the newborn. Indeed lively, friendly and interesting discussions again characterized this very constructive meeting held in ideal warm weather beside Lac Leman. PMID- 11875679 TI - Time-related decay of the benefits of biofeedback therapy. AB - Although immediate results are good to excellent in great majority of patients who undergo biofeedback treatment (BFT) for chronic constipation and fecal incontinence, they tend to loose the benefit over a period of time. The purpose of this study was to evaluate the long-term sustainability of results after successful biofeedback treatment. Two groups of patients who successfully completed BFT at our institution from 1995 to 1997 were created based on the date of completion. The first had a mean follow-up of 35 months and the second group was followed for an average of 12 months. Both groups were questioned as to the presence of constipation and incontinence. The questioning was focused depending on the patient's diagnosis. This information was then compared with the initial BFT results. Overall, all patients were satisfied by the initial BFT results. All patients initially had an excellent or good response to BFT. However, after a mean of 35 months, in the first group, 19 of 22 patients had a near complete regression back to their pre-biofeedback status. In the 14 patients in the second group with mean follow-up of 12 months, 11 had a significant decay in benefits. Only time was a significant factor in the decay of BFT benefits. In conclusion, BFT is highly effective in the treatment of selected patients with complex defecation disorders. Although there is a high initial success rate, there is a clear loss of the immediate benefits over time. Other factors such as dietary habits, pelvic floor exercises, manometry, invasive EMG, and rectal sensation did not correlate with long-term outcomes. The comparison between the two groups reveals a linear model describing the time decay of the benefits of BFT. Based on the linear model, patients may need reevaluation after one year and may benefit from additional BFT. PMID- 11875680 TI - Prospective randomised trial comparing ayurvedic cutting seton and fistulotomy for low fistula-in-ano. AB - The aim of this study was to evaluate the role of ayurvedic setons in the treatment of low fistula-in-ano. One hundred and eight patients were randomised into either conventional fistulotomy (F) or ayurvedic cutting seton insertion (C). Endpoints investigated included time to wound healing and complications of surgery. Post-operative pain scores were measured daily using a visual analog scale. Anal function was compared using a continence score. Pre- and postoperative manometry and ultrasound were also performed. After exclusions, there were 54 patients in group F and 46 in group C. There were no differences in age, sex or follow-up duration between the two groups. Healing time was similar between the groups. Group C reported more pain following operation and on the first 2-4 postoperative days, but both groups experienced the same amount of pain subsequently. In conclusion, chemical seton was more painful than conventional fistulotomy in the first few days following surgery. However, there was no difference in time to wound healing, complications or functional outcome. PMID- 11875681 TI - Long-term results of topical nitroglycerin in the treatment of chronic anal fissures are disappointing. AB - Several treatments for anal fissure offer different responses and risks for complications. Reversible chemical sphincterotomy with topical nitroglycerin is an alternative to surgical sphincterotomy. The aim of this observational prospective study was to compare the effectiveness of topical nitroglycerin with the conventional medical treatment. Forty-three patients with anal fissure were treated: 22 (16 chronic and 6 acute) received 0.25% nitroglycerin topically, and 21 (16 chronic and 5 acute) received conventional treatment. Nitroglycerin or placebo was administered in two daily applications for two weeks. Median follow up was 39 weeks. Healing rate with NTG was 75% for chronic and 83% for acute fissures, but recurrence rate for chronic fissures at 9 months was 67%. Healing rate with placebo for chronic fissures was only anecdotical (1 out of 16 patients). Headache as side effects occurred in 17 cases (77%). In conclusion, both treatments were effective for acute fissures, whereas for chronic ones NTG had a high healing rate but also a high recurrence rate. PMID- 11875682 TI - Which surgical approach for rectocele? A multicentric report from Italian coloproctologists. AB - The most effective surgical technique for rectocele has not yet been clearly established. A retrospective multicentric study was carried out to compare the long-term results of 3 endorectal techniques (Block, Sarles and stapled) and the perineal levatorplasty, alone and in association, in a series of patients with symptomatic rectocele. From January 1992 to December 1999, 2212 patients with defecation disorders were referred to 5 Italian coloproctology units. An anterior rectocele was clinically diagnosed in 1045 patients and confirmed with defecography. On the basis of clinical and radiological parameters, 317 patients (312 women; mean age, 52.4+/-20.1 years) were selected for surgery. Group 1 consisted of 141 patients (136 women; mean age, 50.4+/-18.8 years) who were submitted to endorectal operations. Group 2 consisted of 126 women (mean age, 52.5+/-19.7 years) who received perineal levatorplasty. Finally, 50 women (mean age, 54.3+/-21.9 years) in Group 3 received endorectal operations associated with perineal levatorplasty. A total of 269 patients were followed postoperatively (mean period, 24.2+/-3.1 months, 27.5+/-5.4 months and, 22.8+/-2.8 months, respectively) with the same questionnaire and clinical examination. Three months after surgery, a defecography examination and anorectal manometry were performed in 136 and 132 patients, respectively. Operative time, hospital stay and time to return to work were significantly higher in Group 3 (p<0.001). There was one death in Group 3 due to severe sepsis. Main postoperative complications were: in Group 1, hemorrhage (7.8%, all Sarles), dehiscence of the endorectal suture (5.0%, all Block), distal rectal stenosis (2.1%, 1 stapled, 2 block), and rectovaginal fistula (1.4%, all Sarles); in Group 2, delayed healing of the perineal wound (16.4%); in Group 3 delayed healing of the perineal wound (22.0%), hemorrhage (6%, all Sarles), dehiscence (4.0%), stenosis (2.0%). 17.3% of patients of Group 2 and 22.5% of Group 3 complained of dyspareunia. Postoperative defecography showed a complete absence of the rectocele in 44.1% of patients and reduction of size in the others, without significant differences among the three groups. Manometric pattern was not significantly modified by surgery. Significant symptoms recurred in 5.9% of the patients in Group 1, 6.4% in Group 2, and 5.0% in Group 3. Perineal levatorplasty did not significantly improve obstructed defecation, as it did not allow to excise the rectal mucosal prolapse, and was followed by an high incidence of delayed healing of the perineal wound and dyspareunia. Sarles procedure achieved better control of mucosal prolapse but carried a higher complication rate compared to the others. The association of the perineal levatorplasty with an endorectal technique required significantly longer operative time, and led to a longer hospital stay and time to return to work. In conclusion, the investigated techniques showed different patterns of postoperative complications: bleeding after Sarles, dehiscence after Block, dyspareunia after perineoplasty and fatal gangrene after stapled, but non of them showed a clear superiority over the others in term of clinical or functional results 2 years after surgery. PMID- 11875683 TI - Associations of defecography and physiologic findings in male patients with rectocele. AB - This study evaluated the incidence and physiological findings in male patients with rectoceles. All defecographic studies were evaluated by a single colorectal surgeon. After diagnosis of rectocele in male patients, the patient's history, symptoms, and physiologic tests (anal manometry, pudendal nerve terminal motor latency [PNTML], assessment and electromyography [EMG]) were studied. A prominent rectocele was defined as one that did not empty during defecography and was associated with outlet obstructive syndrome. Forty (17%) rectoceles were diagnosed in 234 male patients with evacuatory disorders who underwent defecography. Rectoceles were anterior in 19 (48%) and posterior in 21 (52%) patients. The main complaint was constipation with difficult defecation in 33 (83%), followed by rectal pain in 5 (13%), rectal prolapse in 1 (3%), and incontinence in 1 (3%). Previous prostatic surgery had been performed in 16 (40%) patients. The mean age and duration of symptoms were 72.4 years (range, 30-88) and 10.3 years (range, 0.5-70), respectively. Excessive straining during evacuation was noted in 73%, unilateral or bilateral pudendal neuropathy in 24.5%, paradoxical puborectalis contraction in 49% and abnormal EMG in 11% of patients. Higher resting pressures with a mean 3.9 cm high pressure zone were noted in 29% of patients. The accompanying findings in defecography were, non relaxing or partially relaxing puborectalis muscle (66%), perineal descent (65%), intussusception (23%), and sigmoidocele (15%). None of the patients underwent surgery for rectocele alone. In conclusion, rectocele is uncommon in males; it rarely appears as an isolated dysfunction as it is often associated with functional disorders of the pelvic floor. There is a frequent association between rectocele and prostatectomy. Clinical significance and therapeutic strategy remain unknown. PMID- 11875684 TI - Bristol scale stool form. A still valid help in medical practice and clinical research. AB - The collection of clinical data concerning bowel habit is always empirical. A more extended use of visual descriptive stool form scales could contribute to a clearer and more standardized reporting of data about bowel function. This could be helpful for both clinical practice and research purposes. PMID- 11875685 TI - The evaluation and physiologic assessment of hemorrhoidal disease: a review. AB - The term hemorrhoids in generally used to describe "symptomatic hemorrhoids". A Medline review of the literature on anatomy, physiology and post-hemorrhoidectomy changes was performed and summarized in this review. PMID- 11875686 TI - Mesh repair of parastomal hernias: new aspects of the Onlay technique. AB - When a hernia becomes symptomatic with pain, obstruction, or mechanical distortion, need for a repair is likely. In this short note are new aspects on the Onlay mesh repair technique of parastomal hernias presented. The satisfactory results achieved in 5 patients are reported. One recurrence required a further successful repair. PMID- 11875687 TI - Chilaiditi's syndrome. Successful surgical correction by colopexy. AB - We describe the case of a patient affected by the Chilaiditi's syndrome, the interposition of the small or large bowel between the inferior face of the diaphragm and the liver, a rare anomaly, often asymptomatic. A 50-year-old man came to our outpatients clinic because of persistent pain in the right abdominal region. Two attempts to perform colonoscopy failed because of the impossibility of passing through the transverse colon. A double contrast enema indicated only sigmoid diverticulitis. Only after abdominal radiography was the abnormal position of the right colon noted. At surgery the right colon was totally intraperitoneal and positioned between the diaphragm and liver, which was smaller than normal. The right colon was repositioned and fixed to the anterior abdominal wall. The postoperative course was uneventful and the right abdominal pain disappeared completely. In conclusion, surgical treatment of Chilaiditi's syndrome may be required in cases of persistent abdominal pain. PMID- 11875689 TI - Conversion of a pelvic pouch to a continent pouch (Kock pouch). PMID- 11875690 TI - A controlled "before-after" study: impact of a clinical guidelines programme and regional cancer network organization on medical practice. AB - A regional cancer network has been set up in the Rhone-Alpes region in France. The aim of the project is to improve the quality of care and to rationalize prescriptions in the network. In this network, we assessed the impact of the implementation of a clinical practice guidelines project by assessing the conformity of practice with the guidelines and comparing this with the conformity in an external matched control group from another French region without a regional cancer network. Four hospitals (private and public) accepted to assess the impact of the clinical practice guidelines on the management of breast and colon cancer in the experimental group and three hospitals (private and public) in the control group. In 1994 and 1996, women with non-metastatic breast cancer (282 and 346 patients in the experimental group, 194 and 172 patients in the control group, respectively) and all new patients with colon cancer (95 and 94 patients in the experimental group, and 89 and 118 patients in the control group, respectively) were selected. A controlled "before-after" study, using institutional medical records of patients with breast and colon cancer. The medical decisions concerning the patients were analyzed to assess their compliance with the clinical practice guidelines. When medical decisions were judged to be non-compliant, we verified if they were based on scientific evidence in a published article, if they were not, the medical decision was classified as having "no convincing supporting scientific evidence". The compliance rates were significantly higher in 1996 than in 1994 in the experimental group; 36% (126 out of 346) vs 12% (34 out of 282) and 46% (56 out of 123) vs 14% (14 out of 103) (P<0.001) for breast and colon cancer, respectively. Whereas, in the control group the compliance rates were the same for the two periods; 7% (12 out of 173) vs 6% (12 out of 194) (P=0.46) and 39% (49 out of 126) vs 32% (31 out of 96), P=0.19. In the experimental group, in 1994, 101 of the 282 medical decisions (36%) and 27 of the 103 (26%) for breast and colon cancer, respectively, were classified as having "no convincing supporting scientific evidence" compare with 72 out of 346 in 1996 (21%) for breast cancer, and 21 of the 123 (17%) for colon cancer P<0.05. Whereas in the control group these results were 106 out of 194 in 1994 (55%) and 90 out of 172 in 1996 (52%), P=0.65 for breast cancer and 28 out of 96 in 1994 (29%) and 30 out of 126 in 1996 (24%), P=0.36 for colon cancer. The development and implementation strategy of the clinical practice guidelines programme for cancer management results in significant changes in medical practice in our cancer network. These results would suggest that introducing guidelines with specific implementation strategy might also increase the compliance rate with the guideline and "evidence-based medicine". PMID- 11875691 TI - Parathyroid hormone related peptide and receptor expression in paired primary prostate cancer and bone metastases. AB - Parathyroid hormone-related peptide is a regulatory protein implicated in the pathogenesis of bone metastases, particularly in breast carcinoma. Parathyroid hormone-related peptide is widely expressed in primary prostate cancers but there are few reports of its expression in prostatic metastases. The aim of this study was to examine the expression of parathyroid hormone-related peptide and its receptor in matched primary and in bone metastatic tissue from patients with untreated adenocarcinoma of the prostate. Eight-millimetre trephine iliac crest bone biopsies containing metastatic prostate cancer were obtained from 14 patients from whom matched primary tumour tissue was also available. Histological grading was performed by an independent pathologist. The cellular location of mRNA for parathyroid hormone-related peptide and parathyroid hormone-related peptide receptor was identified using in situ hybridization with (35)S-labelled probe. Expression of parathyroid hormone-related peptide and its receptor was described as uniform, heterogenous or negative within the tumour cell population. Parathyroid hormone-related peptide expression was positive in 13 out of 14 primary tumours and in all 14 metastases. Receptor expression was evident in all 14 primaries and 12 out of 14 metastases. Co-expression of parathyroid hormone related peptide and parathyroid hormone-related peptide receptor was common (13 primary tumours, 12 metastases). The co-expression of parathyroid hormone-related peptide and its receptor suggest that autocrine parathyroid hormone-related peptide mediated stimulation may be a mechanism of escape from normal growth regulatory pathways. The high frequency of parathyroid hormone-related peptide expression in metastases is consistent with a role in the pathogenesis of bone metastases. PMID- 11875692 TI - Phase II multicentre study of docetaxel plus cisplatin in patients with advanced urothelial cancer. AB - A multicentre phase II trial was undertaken to evaluate the activity and toxicity of docetaxel plus cisplatin as first-line chemotherapy in patients with urothelial cancer. Thirty-eight patients with locally advanced or metastatic transitional-cell carcinoma of the bladder, renal pelvis or ureter received the combination of docetaxel 75 mg m(-2) and cisplatin 75 mg m(-2) on day 1 and repeated every 21 days, to a maximum of six cycles. The median delivered dose intensity was 98% (range 79-102%) of the planned dose for both drugs. There were seven complete responses and 15 partial responses, for and overall response rate of 58% (95% CI, 41-74%). Responses were even seen in three patients with hepatic metastases. The median time to progression was 6.9 months, and the median overall survival was 10.4 months. Two patients who achieved CR status remain free of disease at 4 and 3 years respectively. Grade 3-4 granulocytopenia occurred in 27 patients, resulting in five episodes of febrile neutropenia. There was one toxic death in a patient with grade 4 granulocytopenia who developed acute abdomen. Grade 3-4 thrombocytopenia was rare (one patient). Other grade 3-4 toxicities observed were anaemia (three patients), vomiting (five patients), diarrhoea (four patients), peripheral neuropathy (two patients) and non-neutropenic infections (seven patients). Docetaxel plus cisplatin is an effective and well-tolerated regimen for the treatment of advanced urothelial cancer, and warrants further investigation. PMID- 11875693 TI - Outcome following surgery for colorectal cancer: analysis by hospital after adjustment for case-mix and deprivation. AB - Outcome, adjusted for case-mix and deprivation, in 3200 patients undergoing resection for colorectal cancer in 11 hospitals in Central Scotland between 1991 and 1994 was studied. There were significant differences among individual hospitals in the proportion of elderly (P<0.001) and deprived (P<0.0001) patients, the mode (P=0.007) and stage (P<0.0001) at presentation, and the proportion of patients who underwent apparently curative resection (P<0.001). There were no significant differences in postoperative mortality. Cancer-specific survival at 5 years following apparently curative resection varied from 59 to 76%; cancer-specific survival at 2 years following palliative resection varied from 22 to 44%. The corresponding hazard ratios, adjusted for the above prognostic factors, for patients undergoing apparently curative resection varied among hospitals from 0.58 to 1.32; and the ratios for palliative resection varied from 0.73 to 1.26. This study demonstrates that, after adjustment for variations in case-mix and deprivation, significant differences in outcome among hospitals following resection for colorectal cancer persist. PMID- 11875694 TI - Clinical and immunological assessment of Mycobacterium vaccae (SRL172) with chemotherapy in patients with malignant mesothelioma. AB - The objectives of this study were to determine the toxicity of intratumoural/intrapleural SRL172 in addition to intradermal SRL172 and standard chemotherapy (mitomycin-C, vinblastine and cisplatin) in patients with malignant mesothelioma. Patients received chemotherapy (mitomycin-C: 8 mg m(-2), vinblastine: 6 mg m(-2), cisplatin 50 mg m(-2)) on a 3-weekly basis for up to six courses. IP SRL172 injections were given 3-weekly prior to chemotherapy and escalated in groups of three patients from 1 microg to 1 mg bacilli in 10-fold increments. Patients were also given ID SRL172 at a dose of 1 mg bacilli 4 weekly. Patients were assessed for toxicity after each course of chemotherapy and for response by CT imaging. Immuno-haematological parameters were analyzed pre treatment and 1 month after completion of treatment. There was no dose limiting toxicity with IP SRL172 although there was greater toxicity at the highest dose (n=13). There were six out of 16 partial responses (37.5%). Haemato-immunological parameters, measured in seven patients pre and post-therapy, revealed that response rate correlated with a decrease in platelet count and there was an increase in activation of natural killer cells and a decrease in the percentage of IL-4 producing T cells in all tested patients post-treatment. SRL172 can be given safely into tumour deposits and the pleural cavity in patients with malignant mesothelioma and we have established the dose for phase II testing. PMID- 11875695 TI - Multicentre phase II study of gemcitabine and cisplatin in malignant pleural mesothelioma. AB - Malignant pleural mesothelioma is a notoriously chemoresistant tumour. However, a recent single institution study showed an impressive activity of gemcitabine and cisplatin. Our aim is to investigate the efficacy and toxicity of a gemcitabine and cisplatin combination in selected and chemo-naive patients with histologically proven malignant pleural mesothelioma. METHOD: Gemcitabine 1250 mg m(-2) was administered on day 1 and day 8 and cisplatin 80 mg m(-2) was administered on day 1 in a 3-week cycle with a maximum of six cycles. Response and toxicity evaluations were performed according to WHO and NCIC-CTC criteria. Pathology and radiology were centrally reviewed. Results show that in 25 evaluable patients, four PR were observed (ORR 16%, 95% CI 1-31%). Responses of seven patients were unevaluable. No unexpected toxicity occurred. Time to progression was 6 months (5-7 months) with a median survival from registration of 9.6 months (95% CI 8-12 months). In conclusion this trial excludes with 90% power a response rate of greater than 30% in patients with malignant pleural mesothelioma using a combination of gemcitabine and cisplatin at the proposed dose and schedule. PMID- 11875696 TI - Congou tea drinking and oesophageal cancer in South China. AB - The study from a large hospital-based case-control for 1248 cases with oesophageal cancer and the same number of controls in South China showed that Congou, a grade of Chinese black tea, may protect against cancers of the oesophagus and reduce the risk of a combination of alcohol drinking and smoking (especially smoking), regardless of temperature when drinking. PMID- 11875697 TI - Mobile phones and malignant melanoma of the eye. AB - Recently a four-fold increase in the risk of malignant melanoma of the eye was associated with the use of radiofrequency transmitting devices, including mobile phones in Germany. We contrasted the incidence rates of this rare cancer with the number of mobile phone subscribers in Denmark. We observed no increasing trend in the incidence rate of melanoma, which was in sharp contrast to the exponentially increasing number of mobile phone subscribers starting in the early 1980s. Our study provides no support for an association between mobile phones and ocular melanoma. PMID- 11875698 TI - The importance of full participation: lessons from a national case-control study. AB - Differential participation between cases and controls can lead to biased estimates of risk. However, the effects of participation are often ignored. We report a detailed analysis of locations of residence for participants and non participants in a large, national case-control study of childhood cancer in Great Britain, using the 1991 census. The initial selection of 7669 controls, taken from lists of those registered with a General Practitioner, was representative of the British population in respect to an areal-based index of material deprivation. However, parents of controls agreeing to participate were living in more affluent areas than initially selected controls and their matched 3838 cases. The three components of the deprivation index, persons unemployed, households not owning a car or their home were similarly associated with participation. Other census characteristics, such as proportion of flat dwellers and centrally heated households were also associated with control participation. Population density of the local area was not different between participating controls and their matched cases. However, initially selected controls lived in more urban areas than their cases. Such differences are not unique to this study, as they are an inevitable consequence of incomplete participation. The implications of these differences are discussed, in relation to the difficulty this imposes in the interpretation of studies of disease aetiology. PMID- 11875699 TI - Association of early life factors and acute lymphoblastic leukaemia in childhood: historical cohort study. AB - In a historical cohort study of all singleton live births in Northern Ireland from 1971-86 (n=434,933) associations between early life factors and childhood acute lymphoblastic leukaemia were investigated. Multivariable analyses showed a positive association between high paternal age (> or =35 years) and acute lymphoblastic leukaemia (relative risk=1.49; 95% confidence interval (CI)=0.96- 2.31) but no association with maternal age. High birth weight (> or =3500 g) was positively associated with acute lymphoblastic leukaemia (relative risk=1.66; 95% CI=1.18--2.33). Children of mothers with a previous miscarriage or increased gestation (> or =40 weeks) had reduced risks of ALL (respective relative risks=0.49; 95% CI=0.29--0.80, and 0.67; 95% CI=0.48--0.94). Children born into more crowded households (> or =1 person per room) had substantially lower risks than children born into less crowded homes with also some evidence of a lower risk for children born into homes with three adults (relative risks=0.56; 95% CI=0.35-0.91 and 0.58; 95% CI=0.21-1.61 respectively). These findings indicate that several early life factors, including living conditions in childhood and maternal miscarriage history, influence risk of acute lymphoblastic leukaemia in childhood. PMID- 11875700 TI - Breast cancer incidence in food- vs non-food-producing areas in Norway: possible beneficial effects of World War II. AB - It has been suggested that World War II influenced breast cancer risk among Norwegian women by affecting adolescent growth. Diet changed substantially during the war, and the reduction in energy intake was assumed to be larger in non-food producing than in food-producing municipalities. In the present study, we have looked at the influence of residential history in areas with and without food production on the incidence of breast cancer in a population-based cohort study consisting of 597,906 women aged between 30 and 64 years. The study included 7311 cases of breast cancer, diagnosed between 1964 and 1992. The risk estimates were calculated using a Poisson regression model. The results suggest that residential history may influence the risk of breast cancer, where the suggested advantageous effect of World War II seems to be larger in non-food-producing than in food producing areas. Breast cancer incidence was observed to decline for the post-war cohorts, which is discussed in relation to diet. PMID- 11875701 TI - Ethnic differences in ovulatory function in nulliparous women. AB - African-American women have a long-standing approximately 20% higher breast cancer incidence rate than USA White women under age 40 while rates among Latinas are lower than those of Whites. The reasons for this are not clear, however they may be due to ethnic differences in circulating oestradiol and progesterone levels. In a cross-sectional study, we investigated whether anovulation frequency and circulating serum oestradiol and/or progesterone levels vary among normally cycling nulliparous African-American (n=60), Latina (n=112) and non-Latina White (n=69) women. Blood and urine specimens were collected over two menstrual cycles among healthy 17- to 34-year-old women. Frequency of anovulation was greater among White women (nine out of 63, 14.3%) than African-American women (four out of 56, 7.1%) or Latina women (seven out of 102, 6.9%), although these differences were not statistically significant. African-American women had 9.9% (P=0.26) higher follicular phase oestradiol concentrations than Latina women and 17.4% (P=0.13) higher levels than White women. African-American women also had considerably higher levels of luteal phase oestradiol (vs Latinas, +9.4%, P=0.14; vs Whites, +25.3%, P=0.003) and progesterone (vs Latinas, +15.4%, P=0.07; vs Whites, +36.4%, P=0.002). Latina women were also observed to have higher follicular oestradiol, and luteal oestradiol and progesterone levels than White women (follicular oestradiol: +6.8%, P=0.48; luteal oestradiol: +14.6%, P=0.04; luteal progesterone: +18.2%, P=0.06). These results suggest that exposure to endogenous steroid hormones may be greater for young African-American and Latina women than for Whites. PMID- 11875702 TI - Muscle UCP-3 mRNA levels are elevated in weight loss associated with gastrointestinal adenocarcinoma in humans. AB - The mitochondrial uncoupling proteins-2 and -3 are putative mediators of thermogenesis and energy expenditure. We measured the mRNA levels of uncoupling proteins-2 and -3 in skeletal muscle from 12 gastrointestinal adenocarcinoma patients, of whom six had stable weight and six had lost 2-18 kg, and from six healthy controls undergoing elective surgery. Uncoupling proteins-3 mRNA levels were significantly higher in the muscle of the cancer patients with weight loss (2.2 +/- 0.47 arbitrary units) compared both with controls (0.39 +/- 0.20) and with cancer patients who had not lost weight (0.47 +/- 0.23; P<0.02). Uncoupling proteins-2 mRNA levels did not differ significantly between groups. Elevations in muscle uncoupling proteins-3 activity may enhance energy expenditure and this in turn could contribute to tissue catabolism. PMID- 11875703 TI - Abnormal FHIT expression profiles in cervical intraepithelial neoplastic (CIN) lesions. AB - Abnormal fragile histidine triad transcripts were found in 20-30% of CIN2/3 lesions and 11% of normal cervical biopsies by RT-PCR. Bi-allelic loss of the fragile histidine triad gene and the loss of fragile histidine triad protein expression detectable by immunochemical staining with a polyclonal fragile histidine triad specific antibody was rare. The genomic changes showed no association with the presence of human papillomavirus types which carry high risk for cervical cancer (high risk human papillomavirus) as assessed by a type specific multiplex PCR. The presence of abnormal fragile histidine triad transcripts in a subset of CIN2/3 lesions with no high risk human papillomavirus suggests that this could be an independent risk factor associated with an alternative carcinogenic pathway. PMID- 11875704 TI - Primary screening for cervical cancer precursors by the combined use of liquid based cytology, computer-assisted cytology and HPV DNA testing. AB - Primary screening for cervical cancer precursors has considerably evolved with the introduction of new technology to improve the early detection of disease. The objective of this study was to elaborate a diagnostic pathway integrating liquid based and computer-assisted cytology and human papillomavirus DNA testing to focus screening on women at risk which may be more cost-effective for the healthcare system. A single laboratory analysis was conducted during a 5-month period using liquid-based cytology followed by human papillomavirus DNA testing for women with an abnormal result or with previous abnormal cytology. Human papillomavirus prevalence was estimated by testing 909 consecutive unselected samples. All slides were then rescreened using automated cytologic testing and triaged into a high- or low-score group according to computer results. Of the 8676 slides scanned, 352 had a test result of atypical squamous cells of undetermined significance or worse. Two hundred and ninety-seven (84.3%) samples with an atypical squamous cells of undetermined significance or worse result and 100% of those with detection of high-grade squamous intraepithelial lesions and carcinomas (HSIL+) were triaged into the high-score group. The combination of instrument scores and human papillomavirus results indicated that 51.0% of high score/human papillomavirus-positive cases should be considered as ASCUS+, while 99.6% of low-score/human papillomavirus negative cases remained negative in the final cytologic diagnosis, representing 49.0% of all cases. Of the screened women 89.5% should test negative for human papillomavirus and be reported as such in the final cytologic diagnosis. In conclusion, preliminary results suggest that this diagnostic pathway has the potential to improve primary cervical cancer screening and cost-effectiveness. By using a combination of testing methods to focus screening and clinical attention to cases at risk, it would be possible to lengthen screening intervals for 90% of women and to archive without further review all low-score/human papillomavirus-negative slides, representing 50% of the screening workload. PMID- 11875705 TI - Plasma fibrin D-dimer levels correlate with tumour volume, progression rate and survival in patients with metastatic breast cancer. AB - Plasma levels of D-dimer are elevated in cancer patients. Activation of the extrinsic coagulation system and the fibrinolytic cascade within a tumour is thought to be related with growth, invasion and metastasis. We have investigated the relationship between these markers of fibrin metabolism, standard clinicopathological variables and serum levels of angiogenic cytokines in three cohorts: group A (n=30) consisted of 30 healthy female volunteers, group B (n=23) of consecutive patients with operable breast cancer and group C (n=84) of patients with untreated or progressive metastatic breast cancer. Plasma D-dimers, fibrinogen, IL-6, vascular endothelial growth factor and calculated vascular endothelial growth factor load in platelets are clearly increased in patients with breast cancer. D-dimers were increased in nearly 89% of patients with progressive metastatic disease. The level of D-dimers was positively correlated with tumour load (P<0.0001), number of metastatic sites (P=0.002), progression kinetics (P<0.0001) and the cytokines related to angiogenesis: serum vascular endothelial growth factor (P=0.0016, Spearman correlation=0.285), calculated vascular endothelial growth factor load in platelets (P<0.0001, Spearman correlation=0.37) and serum interleukin-6 (P<0.0001, Spearman correlation=0.59). Similarly increased D-dimer levels were positively correlated with increased fibrinogen levels (P<0.0001, Spearman correlation=0.38). The association between markers of fibrin degradation in patients with progressive breast cancer suggests that the D-dimer level is a clinically important marker for progression and points towards a relation between haemostasis and tumour progression. A role of interleukin-6, by influencing both angiogenesis and haemostasis, is suggested by these observations. PMID- 11875706 TI - Occult axillary lymph node metastases are of no prognostic significance in breast cancer. AB - The significance of occult metastases in axillary lymph nodes in patients with carcinoma of the breast is controversial. Additional sections were cut from the axillary lymph nodes of 477 women with invasive carcinoma of the breast, in whom no metastases were seen on initial assessment of haematoxylin and eosin stained sections of the nodes. One section was stained with haematoxylin and eosin, and one using immunohistochemistry with two anti-epithelial antibodies (CAM5.2 and HMFG2). Occult metastases were found in 60 patients (13%). The median follow-up was 18.9 years with 153 breast cancer related deaths. There was no difference in survival between those with and those without occult metastases. Multivariate analysis, however, showed that survival was related to tumour size and histological grade. This node-negative group was compared with a second group of 202 patients who had one involved axillary node found on initial assessment of the haematoxylin and eosin sections; survival was worse in the patients in whom a nodal metastasis was found at the time of surgery. Survival was not related to the size of nodal metastases in the occult metastases and single node positive groups. Some previous studies have found a worse prognosis associated with occult metastases on univariate analysis, but the evidence that it is an independent prognostic factor on multivariate analysis is weak. We believe that the current evidence does not support the routine use of serial sections or immunohistochemistry for the detection of occult metastases in the management of lymph node negative patients, but that the traditional factors of histological grade and tumour size are useful. PMID- 11875707 TI - Cyclin A is a prognostic indicator in early stage breast cancer with and without tamoxifen treatment. AB - Overexpression of G1-S regulators cyclin D1 or cyclin A is frequently observed in breast cancer and is also to result in ligand-independent activation of oestrogen receptor in vitro. This might therefore, provide a mechanism for failure of tamoxifen treatment. We examined by immunohistochemical staining the effect of deregulation of these, and other cell cycle regulators on tamoxifen treatment in a group of 394 patients with early stage breast cancer. In univariate analysis, expression of cyclin A, Neu, Ki-67 index, and lack of OR expression were significantly associated with worse prognosis. When adjusted by the clinical model (for lymph node status, age, performance status, T-classification, grade, prior surgery, oestrogen receptor status and tamoxifen use), only overexpression of cyclin A and Neu were significantly associated with worse prognosis with hazard ratios of, respectively, 1.709 (P=0.0195) and 1.884 (P=0.0151). Overexpression of cyclin A was found in 86 out of the 201 OR-positive cases treated with tamoxifen, and was the only independent marker associated with worse prognosis (hazard ratio 2.024, P=0.0462). In conclusion, cyclin A is an independent predictor of recurrence of early stage breast cancer and is as such a marker for response in patients treated with tamoxifen. PMID- 11875708 TI - S100A4 (p9Ka) protein in colon carcinoma and liver metastases: association with carcinoma cells and T-lymphocytes. AB - The presence of the EF-hand-calcium-binding protein S100A4 in the carcinoma cells of the primary tumour is associated with a shorter survival time of a group of breast cancer patients. In colon cancer, primary tumours as well as metastases to the liver can be studied. Here we show, using quantitative PCR applied to RNA from 24 normal colon, four liver tissues, 24 colon carcinoma specimens, and 24 livers containing colonic carcinoma metastases, that the level of S100A4 mRNA was significantly higher in the carcinomas compared to normal specimens (Mann-Whitney U-test, P=0.05), and in liver metastases compared to carcinoma specimens (P=0.039). The latter comparison included seven liver metastases and their matched primary carcinomas (P<0.001) from the same patient. In situ hybridization and immunocytochemistry techniques have localized S100A4 to both carcinoma cells and lymphocytes in the malignant specimens. The percentage of specimens stained for S100A4 in the epithelial cells is significantly higher for those isolated from carcinomas and metastases than from the corresponding normal tissue, and from metastases than from corresponding carcinoma (Fisher Exact text, P<0.0016, P=0.04, respectively). In most specimens, S100A4 is present in clusters of T lymphocytes and this distribution is also found in the lymphoid, uninflamed appendix. PMID- 11875709 TI - Prognostic impact of matched preoperative plasma and serum VEGF in patients with primary colorectal carcinoma. AB - In serum, the major part of vascular endothelial growth factor derives from in vitro degranulation of granulocytes and platelets. Therefore, plasma may be preferred for vascular endothelial growth factor measurements. However, which specimen is the best predictor of survival is still debated. The present study analyzed the prognostic value of matched preoperative serum and plasma vascular endothelial growth factor concentrations in patients with colorectal cancer. To establish the reference range among healthy people, vascular endothelial growth factor was analyzed in 50 matched EDTA-plasma and serum samples from healthy blood donors. Preoperatively, in 524 patients with colorectal cancer, matched plasma and serum vascular endothelial growth factor concentrations were analyzed. In the colorectal cancer patients, the median plasma vascular endothelial growth factor concentration (44 pg ml(-1)) was significantly (P=0.01) higher than the median plasma vascular endothelial growth factor concentration (30 pg ml(-1)) in the healthy blood donors. In serum, no significant (P=0.30) difference in the median vascular endothelial growth factor concentration was found between colorectal cancer patients (268 pg ml(-1)) and healthy blood donors (220 pg ml( 1)). The preoperative vascular endothelial growth factor concentrations were dichotomized by the 95th percentile of the healthy blood donors (plasma=112 pg ml(-1), serum=533 pg ml(-1)). In univariate survival analyses, both high plasma vascular endothelial growth factor (>112 pg ml(-1)) and high serum vascular endothelial growth factor (>533 pg ml(-1)) predicted a reduced survival. In multivariate survival analyses, high serum vascular endothelial growth factor (>533 pg ml(-1)) independently predicted a reduced survival (HR=1.65, P=0.015), while high plasma vascular endothelial growth factor (>112 pg ml(-1)) did not (HR=1.27, P=0.23). This study indicates that preoperative serum vascular endothelial growth factor apparently is a better predictor of overall survival than the preoperative plasma vascular endothelial growth factor. PMID- 11875710 TI - Role of beta3-adrenergic receptors in the action of a tumour lipid mobilizing factor. AB - Induction of lipolysis in murine white adipocytes, and stimulation of adenylate cyclase in adipocyte plasma membranes, by a tumour-produced lipid mobilizing factor, was attenuated by low concentrations (10(-7)--10(-5)M) of the specific beta3-adrenoceptor antagonist SR59230A. Lipid mobilizing factor (250 nM) produced comparable increases in intracellular cyclic AMP in CHOK1 cells transfected with the human beta3-adrenoceptor to that obtained with isoprenaline (1 nM). In both cases cyclic AMP production was attenuated by SR59230A confirming that the effect is mediated through a beta3-adrenoceptor. A non-linear regression analysis of binding of lipid mobilizing factor to the beta3-adrenoceptor showed a high affinity binding site with a Kd value 78 +/- 45 nM and a B(max) value (282 +/- 1 fmole mg protein(-1)) comparable with that of other beta3-adrenoceptor agonists. These results suggest that lipid mobilizing factor induces lipolysis through binding to a beta3-adrenoceptor. PMID- 11875711 TI - Local hypoxia is produced at sites of intratumour injection. AB - Intratumour injection, commonly used for gene or drug delivery but also associated with needle biopsy or insertion of invasive measuring devices, may damage tumour microvessels. To examine this possibility, SCCVII tumours grown subcutaneously in C3H mice were injected with a 26 gauge needle containing 0.1 ml of the fluorescent dye Hoechst 33342 to label cells lining the track of the needle. Hoechst-labelled cells sorted from these tumours were more sensitive to killing by hypoxic cell cytotoxins (tirapazamine, RSU-1069) and less sensitive to damage by ionizing radiation. Hoechst-labelled cells also bound the hypoxia marker pimonidazole when given by i.p. injection. Intratumour injection transiently increased hypoxia from 18 to 70% in the tumour cells adjacent to the track of the needle. The half-time for return to pre-treatment oxygenation was about 30 min; oxygenation of tumour cells along the track had recovered by 20 h after intratumour injection. This effect could have significant implications for intratumour injection of drugs, cytokines or vectors that are affected by the oxygenation status of the tumour cells as well as potential effects on biodistribution via local microvasculature. PMID- 11875712 TI - Prerequisites for effective adenovirus mediated gene therapy of colorectal liver metastases in the rat using an intracellular neutralizing antibody fragment to p21-Ras. AB - Ras mutations are present in 40-50% of colorectal cancers. Inactivating this oncogene may therefore reduce proliferation capacity. In order to target ras we studied the transduction efficacy and anti tumour activity of an adenoviral vector expressing an intracellular, neutralizing single chain antibody to p21-ras (Y28). In in vitro studies transfection levels of the K-ras mutated rat colon carcinoma cell line CC531 were studied using the LacZ marker gene. In our in vivo liver metastases model different routes of administration were evaluated to determine which regimen resulted in the best transfection levels and tumour responses: intravenous injection, intratumoural injection, isolated liver perfusion, or hepatic artery infusion. CC531 cells are readily transfected in vitro, resulting in significant inhibition of tumour cell proliferation by the Y28 construct. Intravenous injection did not result in any measurable transfection. Intratumoural injection resulted only in the transfection of tumour cells along the needle track. IHP as well as single HAI achieved low transfection levels of tumour tissue. Expression of Y28 was demonstrated in tumours after IT injection, HAI and IHP. Whereas, repeated HAI's clearly achieved expression in and around tumour associated vessels. Only five times repeated HAI's with Y28 resulted in a tumour response: in all animals tumour growth was inhibited, and in three rats out of eight a complete regression of the liver tumours was observed. PMID- 11875713 TI - Ciprofloxacin induces apoptosis and inhibits proliferation of human colorectal carcinoma cells. AB - Efficacy of chemotherapy in advanced stages of colorectal tumours is limited. The quinolone antibiotic ciprofloxacin was recently shown to inhibit growth and to induce apoptosis in human bladder carcinomas cells. We investigated the effect of ciprofloxacin on colon carcinoma lines in vitro. CC-531, SW-403 and HT-29 colon carcinoma and HepG2 hepatoma cells (control cells) were exposed to ciprofloxacin. Proliferation was assessed by bromodeoxyuridine-incorporation into DNA and apoptosis was measured by flow cytometry after propidium iodide or JC-1 staining. Expression of anti-apoptotic Bcl-2 and pro-apoptotic Bax was analyzed by semiquantitative Western blot analysis and activity of caspases 3, 8 and 9 by substrate-cleavage assays. Ciprofloxacin suppressed DNA synthesis of all colon carcinoma cells time- and dose-dependently, whereas the hepatoma cells remained unaffected. Apoptosis reached its maximum between 200 and 500 microg ml(-1). This was accompanied by an upregulation of Bax and of the activity of caspases 3, 8 and 9, and paralleled by a decrease of the mitochondrial membrane potential. Ciprofloxacin decreases proliferation and induces apoptosis of colon carcinoma cells, possibly in part by blocking mitochondrial DNA synthesis. Therefore, qualification of ciprofloxacin as adjunctive agent for colorectal cancer should be evaluated. PMID- 11875714 TI - Interferon-alpha resistance in renal carcinoma cells is associated with defective induction of signal transducer and activator of transcription 1 which can be restored by a supernatant of phorbol 12-myristate 13-acetate stimulated peripheral blood mononuclear cells. AB - Therapy of selected human malignancies with interferon-alpha is widely accepted but often complicated by the emergence of interferon-alpha resistance. Interferon is a pleiotropic cytokine with antiproliferative, antitumour, antiviral and immunmodulatory effect; it signals through the Jak-STAT signal transduction pathway where signal transducer and activator of transcription 1 plays an important role. Here we report both, a lack of signal transducer and activator of transcription induction in interferon-alpha resistant renal cell carcinoma cells and signal transducer and activator of transcription 1 reinduction of phorbol 12 myristate 13-acetate-stimulated peripheral blood mononuclear cells supernatant. Preliminary experiments on the identification of the molecules that reinducing signal transducers and activators of transcription 1 indicate that interferon gamma may be the responsible candidate cytokine, but several others may be involved as well. This work provides the basis for therapeutic strategies directed at the molecular modulation of interferon-alpha resistance in human neoplasms. PMID- 11875715 TI - Targeting the EGF receptor in ovarian cancer with the tyrosine kinase inhibitor ZD 1839 ("Iressa"). AB - The modulating effects of the orally active epidermal growth factor receptor specific tyrosine kinase inhibitor ZD 1839 ("Iressa") on cell growth and signalling were evaluated in four ovarian cancer cell lines (PE01, PE04, SKOV-3, OVCAR-5) that express the epidermal growth factor receptor, and in A2780, which is epidermal growth factor receptor-negative. Transforming growth factor-alpha stimulated growth was completely inhibited by concentrations of ZD 1839 > or =0.3 microM in the epidermal growth factor receptor-expressing cell lines, as were transforming growth factor-alpha stimulated phosphorylation of the epidermal growth factor receptor and downstream components of the MAP kinase and PI-3 kinase signalling cascades. Growth inhibition in the absence of added transforming growth factor-alpha was also observed which could be consistent with suppression of action of autocrine epidermal growth factor receptor-activating ligands by ZD 1839. In support of this, transforming growth factor-alpha, EGF and amphiregulin mRNAs were detected by RT-PCR in the epidermal growth factor receptor-expressing cell lines. ZD 1839 inhibited growth of the PE04 ovarian cancer xenograft at 200 mg kg(-1)day(-1). These data lend further support to the view that targeting the epidermal growth factor receptor in ovarian cancer could have therapeutic benefit. PMID- 11875716 TI - Src tyrosine kinase augments taxotere-induced apoptosis through enhanced expression and phosphorylation of Bcl-2. AB - Activation of Src, which has an intrinsic protein tyrosine kinase activity, has been demonstrated in many human tumours, such as colorectal and breast cancers, and is closely associated with the pathogenesis and metastatic potential of these cancers. In this study, we have examined the effect of activated Src on the sensitivity to taxotere, an anticancer drug targeting microtubules, using v-src transfected HAG-1 human gall bladder epithelial cells. As compared with parental HAG-1 cell line, v-src-transfected HAG/src3-1 cells became 5.9 and 7.0-fold sensitive to taxotere for 2 and 24-h exposure, respectively. By contrast, HAG-1 cells transfected with activated Ras, which acts downstream of Src, acquired approximately 2.5- approximately 4.8-fold taxotere resistance. The taxotere sensitivity in HAG/src3-1 cells was reversed, if not completely, by herbimycin A, a specific inhibitor of Src family protein tyrosine kinase, indicating that Src protein tyrosine kinase augments sensitivity to taxotere. Treatment of HAG/src3-1 cells with taxotere resulted in phosphorylation of Bcl-2 and subsequent induction of apoptotic cell death, whereas neither Bcl-2 phosphorylation nor apoptosis occurred in parental or c-H-ras-transfected HAG-1 cells. Interestingly, the Bcl-2 protein is overexpressed in v-src-transfected cell line, compared to those in parental or Ras-transfected cell line. Treatment of HAG/src3-1 cells with herbimycin A significantly reduced the expression and phosphorylation of Bcl-2, and abrogated taxotere-induced apoptosis, suggesting a potential role for Src protein tyrosine kinase in the taxotere-induced apoptotic events. H-7, a protein kinase C inhibitor and wortmannin, a phosphatidylinositol-3 kinase (PI-3 kinase) inhibitor, neither altered taxotere sensitivity nor inhibited taxotere-induced apoptosis in these cells. These data indicate that the ability of activated Src to increase taxotere sensitivity would be mediated by apoptotic events occurring through Src to downstream signal transduction pathways toward Bcl-2 phosphorylation, but not by activated Ras, PI-3 kinase or protein kinase C. PMID- 11875717 TI - Mitomycin C in combination with radiotherapy as a potent inhibitor of tumour cell repopulation in a human squamous cell carcinoma. AB - The potential of Mitomycin C in combination with fractionated irradiation to inhibit tumour cell repopulation of a fast growing squamous cell carcinoma after fractionated radiotherapy was investigated in vivo. A rapidly growing human squamous cell carcinoma (FaDu(dd)) was used for the study. For experiments, NMRI (nu/nu) mice with subcutaneously growing tumours were randomly allocated to no treatment, Mitomycin C, fractionated irradiation (ambient: 11 x 4.5 Gy in 15 days), or fractionated irradiation combined with Mitomycin C. Graded top up doses (clamped blood flow: 0-57 Gy) were given at day 16, 23, 30 or 37. End point of the study was the time to local tumour progression. Data were examined by multiple regression analysis (Cox). Mitomycin C alone resulted in a median time to local tumour progression of 23 (95% confidence limits: 17-43) days, fractionated irradiation in 31 (25-35) days and combined Mitomycin C plus fractionated irradiation in 65 (58-73) days (P=0.02). Mitomycin C decreased the relative risk of local recurrence by 94% (P<<0.001) equivalent to 31.7 Gy top up dose. Repopulation accounted for 1.33 (0.95-1.72) Gy per day top up dose after fractionated irradiation alone and for 0.68 (0.13-1.22) Gy per day after fractionated irradiation+Mitomycin C (P=0.018). Mitomycin C significantly reduces the risk of local recurrence and inhibits tumour cell repopulation in combination with fractionated irradiation in vivo in the tested tumour model. PMID- 11875719 TI - Schedule-dependent response of neuroblastoma cell lines to combinations of etoposide and cisplatin. AB - The growth inhibitory effects of cisplatin and etoposide on neuroblastoma cell lines were investigated in several scheduled combinations. Results were analyzed using median effect and combination index analyses. In all schedules in which cisplatin was administered prior to etoposide a synergistic effect was observed. Conversely, an antagonistic effect was seen in all schedules where etoposide was administered before cisplatin. PMID- 11875718 TI - Mxi1 inhibits the proliferation of U87 glioma cells through down-regulation of cyclin B1 gene expression. AB - Mxi1 is a Mad family member that plays a role in cell proliferation and differentiation. To test the role of Mxi1 on tumorigenesis of glioma cells we transfected a CMV-driven MXI1 cDNA in U87 human glioblastoma cells. Two clones were isolated expressing MXI1 levels 18- and 3.5-fold higher than wild-type U87 cells (clone U87.Mxi1.14 and U87.Mxi1.22, respectively). In vivo, U87.Mxi1.14 cells were not tumorigenic in nude mice and delayed development of tumours was observed with U87.Mxi1.22 cells. In vitro, the proliferation rate was partially and strongly inhibited in U87.Mxi1.22 and U87.Mxi1.14 cells respectively. The cell cycle analysis revealed a relevant accumulation of U87.Mxi1.14 cells in the G(2)/M phase. Interestingly, the expression of cyclin B1 was inhibited to about 60% in U87.Mxi1.14 cells. This inhibition occurs at the transcriptional level and depends, at least in part, on the E-box present on the cyclin B1 promoter. Consistent with this, the endogenous Mxi1 binds this E-box in vitro. Thus, our findings indicate that Mxi1 can act as a tumour suppressor in human glioblastomas through a molecular mechanism involving the transcriptional down-regulation of cyclin B1 gene expression. PMID- 11875720 TI - Overexpression of the p53-inducible brain-specific angiogenesis inhibitor 1 suppresses efficiently tumour angiogenesis. AB - The brain-specific angiogenesis inhibitor 1 gene has been isolated in an attempt to find fragments with p53 "functional" binding sites. As reported herein and by others, brain-specific angiogenesis inhibitor 1 expression is present in some normal tissues, but is reduced or lost in tumour tissues. Such data and its particular structure prompted the hypothesis that brain-specific angiogenesis inhibitor 1 may act as a mediator in the local angiogenesis balance. We herein demonstrate that brain-specific angiogenesis inhibitor 1 over-expression suppresses tumour angiogenesis, delaying significantly the human tumour growth in immunodeficient mice. The inhibitory effect of brain-specific angiogenesis inhibitor 1 was documented using our intravital microscopy system, strongly implicating brain-specific angiogenesis inhibitor 1 as a mediator in the control of tumour angiogenesis. In contrast, in vitro tumour cell proliferation was not inhibited by brain-specific angiogenesis inhibitor 1 transfection, whereas some level of cytotoxicity was assessed for endothelial cells. Immunohistochemical analysis of tumour samples confirmed a reduction in the microvessel density index in brain-specific angiogenesis inhibitor 1-overexpressing tumours. At messenger level, moderate changes could be detected, involving the down-regulation of vascular endothelial growth factor and collagenase-1 expression. Furthermore, brain-specific angiogenesis inhibitor 1 expression that was lost in a selection of human cancer cell lines could be restored by wild-type p53 adenoviral transfection. Brain-specific angiogenesis inhibitor 1 should be considered for gene therapy and development of efficient drugs based on endogenous antiangiogenic molecules. PMID- 11875722 TI - Beatson International Cancer conference: genomic regulation and cancer. PMID- 11875723 TI - Phase II study of bi-weekly administration of paclitaxel and cisplatin in patients with advanced oesophageal cancer. AB - In a phase I study we demonstrated the feasibility of a bi-weekly combination of paclitaxel 180 mg x m(-2) with cisplatin 60 mg x m(-2). In this study we further assessed toxicity and efficacy of this schedule in the treatment of advanced cancer of the oesophagus or the gastro-oesophageal junction. Patients received paclitaxel 180 mg x m(-2) administered over 3 h followed by a 3-h infusion of cisplatin 60 mg x m(-2). Patients were retreated every 2 weeks unless granulocytes were <0.75x10(9) or platelets <75x10(9). Patients were evaluated after three and six cycles and responding patients received a maximum of eight cycles. Fifty-one patients were enrolled into the study. The median age was 56 years (range 32-78). WHO performance status were: 0 (19 patients); 1 (29 patients); 2 (three patients). All patients received at least three cycles of chemotherapy and all were evaluable for toxicity and response. Haematological toxicity consisted of uncomplicated neutropenia grade 3 in 39% and grade 4 in 31% of patients. Five patients (10%) were hospitalised, three patients because of treatment related complications and two patients because of infections without neutropenia. Sensory neurotoxicity was the predominant non-haematological toxicity; grade 1 and 2 neurotoxicity was observed in 43 and 20% of patients, respectively. Response evaluation in 51 patients with measurable disease: complete response 4%, partial response 39%, stable disease 43% and progressive disease in 14% of the patients. The median duration of response was 8 months. The median survival for all patients was 9 (range 2-29+) months and the one-year survival rate was 43%. Four patients who received additional local treatment (two patients surgery and two patients radiotherapy) are still disease free after a follow-up of 20-29 months. This bi-weekly treatment of paclitaxel and cisplatin is well tolerated by patients with advanced oesophageal cancer. The toxicity profile of this regimen compares favourable to that of previously used cisplatin- and paclitaxel-based regimens. Trials are underway evaluating this bi-weekly regimen in a neo-adjuvant setting. PMID- 11875724 TI - Influence of pathological tumour variables on long-term survival in resectable gastric cancer. AB - Although tumour stage and nodal status are established prognostic factors for resectable gastric cancer, the relative importance of other pathological characteristics remains unclear. This study reports univariate and multivariate analyses of the prognostic value of various pathological and staging factors based on 324 patients entered into the MRC randomised surgical trial for gastric cancer. In the univariate analysis tumour stage, nodal status, UICC clinical stage, number of involved nodes, WHO predominant type, mixed Lauren type, Ming type, tumour differentiation, lymphocytic and tumour stromal eosinophilic infiltration were all found to have a significant impact on survival (logrank test, 5% level). In the multivariate analysis, UICC clinical stage and eosinophilic infiltration were found to have a significant influence. Risk of death increased for UICC stage II and III patients (Hazard Ratio for stage II compared to stage I=2.0, 95% Confidence Interval (CI) 1.4-2.9; Hazard Ratio for stage III compared to stage I=3.5, 95% CI 2.5-4.8). Patients with numerous eosinophils had a lower risk of death than those with none (Hazard Ratio=0.5, 95% CI 0.3-0.8). This association between survival and eosinophilic infiltration merits further study. PMID- 11875725 TI - A phase II trial of the vitamin D analogue Seocalcitol (EB1089) in patients with inoperable pancreatic cancer. AB - Inoperable cancer of the exocrine pancreas responds poorly to most conventional anti-cancer agents, and new agents are required to palliate this disease. Seocalcitol (EB1089), a vitamin D analogue, can inhibit growth, induce differentiation and induce apoptosis of cancer cell lines in vitro and can also inhibit growth of pancreatic cancer xenografts in vivo. Thirty-six patients with advanced pancreatic cancer received once daily oral treatment with seocalcitol with dose escalation every 2 weeks until hypercalcaemia occurred, following which patients continued with maintenance therapy. The most frequent toxicity was the anticipated dose-dependent hypercalcaemia, with most patients tolerating a dose of 10-15 microg per day in chronic administration. Fourteen patients completed at least 8 weeks of treatment and were evaluable for efficacy, whereas 22 patients were withdrawn prior to completing 8 weeks' treatment and in 20 of these patients withdrawal was due to clinical deterioration as a result of disease progression. No objective responses were observed, with five of 14 patients having stable disease in whom the duration of stable disease was 82-532 days (median=168 days). The time to treatment failure (n=36) ranged from 22 to 847 days, and with a median survival of approximately 100 days. Seocalcitol is well tolerated in pancreatic cancer but has no objective anti-tumour activity in advanced disease. Further studies are necessary to determine if this agent has any cytostatic activity in this malignancy in minimal disease states. PMID- 11875727 TI - Sequential or alternating administration of docetaxel (Taxotere) combined with FEC in metastatic breast cancer: a randomised phase II trial. AB - The aim of this study, using a Fleming single-stage design, was to explore the efficacy and safety of Taxotere 100 x mg x m(-2) docetaxel and FEC 75 cyclophosphamide 500 mg x m(-2), fluorouracil 500 x mg x m(-2) and epirubicin 75 mg x m(-2), in alternating and sequential schedules for the first-line treatment of metastatic breast cancer. One hundred and thirty-six women were randomly allocated, to one of three treatment regimens: DTX 100 plus FEC 75, alternated for eight courses (ALT); four courses of DTX 100 followed by four courses of FEC 75 (SEQ T); or four courses of FEC 75 followed by four courses of DTX 100 (SEQ F). One hundred and thirty-one women were evaluable for tumour response. Although the treatment outcome was equivalent in the two sequential arms and the alternating regimen (P=0.110, not significant), the response rate was less encouraging in the SEQ F arm (52.3%) than in the other two arms (71.1% for ALT and 70.5% for SEQ T), in which docetaxel was administered first. Time to progression was similar in the ALT, SEQ T and SEQ F arms (9.5, 9.3 and 10.4 months respectively). Grade 3-4 neutropenia was observed in nearly all patients; febrile neutropenia occurred in 9% (ALT), 16% (SEQ T) and 2% (SEQ F) of patients. Few patients (< or =9%) developed grade 3-4 non-haematological toxicities. Relative dose intensity was 97-99% for all regimens. All treatment regimens were active and well tolerated. PMID- 11875726 TI - Pre-treatment nomogram for biochemical control after neoadjuvant androgen deprivation and radical radiotherapy for clinically localised prostate cancer. AB - Phase III studies have demonstrated the clinical benefit of adding neo-adjuvant androgen deprivation to radical radiotherapy for clinically localised prostate cancer. We have developed a nomogram to describe the probability of PSA control for patients treated in this way. Five hundred and seventeen men with clinically localised prostate cancer were treated with 3-6 months of neo-adjuvant androgen deprivation and radical radiotherapy (64Gy in 32#) between 1988 and 1998. Median presenting PSA was 20 ng x ml(-1), and 56% of patients had T3/4 disease. Multivariate analysis of pre-treatment factors was performed, and a nomogram developed to describe PSA-failure-free survival probability. At a median follow up of 44 months, 233 men had developed PSA failure. Presenting PSA, histological grade and clinical T stage were all highly predictive of PSA failure on multivariate analysis. The nomogram score for an individual patient is given by the summation of PSA (<10=0, 10-19=16, 20-49=44, > or =50=100), grade (Gleason 2 4=0, 5-7=44, 8-10=81) and T stage (T1/2=0, T3/4=35). For a nomogram score of 0, 50, 100 and 150 points the 2 year PSA control rate was 93, 87, 75 and 54%, and the 5 year PSA control rate was 82, 67, 44 and 18%. These results are comparable to those using surgery or higher doses of radical radiotherapy alone. The nomogram illustrates the results of multivariate analysis in a visually-striking way, and facilitates comparisons with other treatment methods. PMID- 11875729 TI - Low prevalence of Epstein-Barr virus in incident gastric adenocarcinomas from the United Kingdom. AB - Epstein-Barr virus has been associated with a proportion of typical gastric adenocarcinomas. Here we report that the prevalence of Epstein-Barr virus in gastric adenocarcinomas from the United Kingdom is one of the lowest in the World. Gastric adenocarcinoma is another tumour whose association with Epstein Barr virus varies with the population studied. PMID- 11875728 TI - Randomised beta-carotene supplementation and incidence of cancer and cardiovascular disease in women: is the association modified by baseline plasma level? AB - In a nested case-control study of 513 women with cancer; 130 with cardiovascular disease and equal numbers of controls, we found no effect of randomised beta carotene on risk of cancer or cardiovascular disease within any quartile of baseline plasma beta-carotene, nor was there a trend across quartiles (P for trend 0.15 and 0.62, respectively). PMID- 11875730 TI - Prevalence and determinants of human papillomavirus genital infection in men. AB - Four-hundred-forty-five husbands of women with invasive cervical carcinoma, 165 of women with in situ cervical cancer, and 717 of control women (age range 19-82 years) were interviewed and a sample of exfoliated cells from the penis obtained in seven case-control studies conducted by the International Agency for Research on Cancer. The characteristics of human papillomavirus-positive and human papillomavirus-negative husbands were compared using odds ratios and 95% confidence intervals. Thirteen per cent of the husbands of control women, 18% of the husbands of women with invasive cervical carcinoma, and 21% of the husbands of in situ cervical carcinoma women were positive for penile human papillomavirus DNA. Human papillomavirus 16 was detected in 45 husbands, human papillomavirus 18, 31 or 33 in 19, and human papillomavirus 6/11 in 6, but the majority of human papillomavirus infection (158) was with other or unspecified human papillomavirus types. The same human papillomavirus type was seldom identified in both husband and wife. The strongest variation in penile human papillomavirus infection was by country, with percentages among the husbands of control women ranging between 3% in Spain and 39% in Brazil. Having had over 50 lifetime sexual partners, compared with only one, was associated with an odds ratio of 2.3. PMID- 11875731 TI - Diet and ovarian cancer risk: a case-control study in China. AB - This case-control study, conducted in Zhejiang, China during 1999-2000, investigated whether dietary factors have an aetiological association with ovarian cancer. Cases were 254 patients with histologically confirmed epithelial ovary cancer. The 652 controls comprised 340 hospital visitors, 261 non-neoplasm hospital outpatients without long-term diet modifications and 51 women recruited from the community. A validated food frequency questionnaire was used to measure the habitual diet of cases and controls. The risks of ovarian cancer for the dietary factors were assessed by adjusted odds ratios based on multivariate logistic regression analysis, accounting for potential confounding demographic, lifestyle, familial factors and hormonal status, family ovarian cancer history and total energy intake. The ovarian cancer risk declined with increasing consumption of vegetables and fruits but vice versa with high intakes of animal fat and salted vegetables. The adjusted upper quartile odds ratio compared to the lower quartile was 0.24 (0.1-0.5) for vegetables, 0.36 (0.2-0.7) for fruits, 4.6 (2.2-9.3) for animal fat and 3.4 (2.0-5.8) for preserved (salted) vegetables with significant dose-response relationship. The risk of ovarian cancer also appeared to increase for those women preferring fat, fried, cured and smoked food. PMID- 11875732 TI - A follow-up study of breast and other cancers in families of an unselected series of breast cancer patients. AB - The cancer experience among relatives of an unselected cohort of 402 breast cancer patients was previously reported. Cases and their first degree relatives were flagged at the National Health Service Central Register for continuous notification of cancer registrations and deaths. More than 10 years of follow-up data have been analysed to update cancer risks overall and to estimate breast cancer risk in relatives prospectively according to family history at the time of breast cancer diagnosis in the index case. Significant excesses of breast cancer (RR 2.24, P<0.0001), prostate cancer (RR 1.71, P=0.039) and bone sarcoma (RR 6.564, P=0.042) overall and soft tissue sarcoma in mothers only (RR 15.44, P=0.001) were found. There was no excess of any other cancer, including ovarian. High breast cancer risk in relatives was associated with young age at diagnosis in the index (index <40 years at diagnosis, RR in relatives 3.76, P=0.004). Prospective risk of breast cancer was higher in relatives of index patients who had an affected first degree relative at the time of their diagnosis (no family history, RR 1.87, P=0.012; with a family history, RR 3.72, P=0.015). These prospective risk estimates are valuable in advising relatives of newly diagnosed breast cancer patients. PMID- 11875733 TI - Differential effects of reproductive factors on the risk of pre- and postmenopausal breast cancer. Results from a large cohort of French women. AB - The aim of this study was to obtain a better understanding of the role of hormonal factors in breast cancer risk and to determine whether the effect of reproductive events differs according to age at diagnosis. It analysed the effect of age at menarche, age at first full-term pregnancy, number of full-term pregnancies and number of spontaneous abortions both on the overall risk of breast cancer and on its pre- or postmenopausal onset, using the data on 1718 breast cancer cases, obtained from a large sample of around 100000 French women participating in the E3N cohort study. The results provide further evidence that the overall risk of breast cancer increases with decreasing age at menarche, increasing age at first pregnancy and low parity. No overall effect of spontaneous abortions was observed. The effect of these reproductive factors differed according to menopausal status. Age at menarche had an effect on premenopausal breast cancer risk, with a decrease in risk with increasing age of 7% per year (P<0.05). Compared to those who had their first menstrual periods at 11 or before, women experiencing menarche at 15 or after had an RR of 0.66 (95% CI 0.45-0.97) in the premenopausal group. Age at first full-term pregnancy had an effect on both pre- and postmenopausal breast cancer risk, with significant tests showing increasing risk per year of increasing age (P=0.001 and P<0.05 respectively). A first full-term pregnancy above age 30 conveyed a risk of 1.63 (95% CI 1.12-2.38) and 1.35 (95% CI 1.02-1.78) in the pre- and postmenopausal groups respectively. A protective effect of high parity was observed only for postmenopausal breast cancer risk (P for trend test =0.001), with point estimates of 0.79 (95% CI 0.60-1.04), 0.69 (95% CI 0.54-0.88), 0.66 (95% CI 0.51-0.85) and 0.64 (95% CI 0.48-0.86) associated to a one, two, three and four or more full term pregnancies. A history of spontaneous abortion had no significant effect on the risk of breast cancer diagnosed before or after menopause. Our results suggest that reproductive events have complex effects on the risk of breast cancer. PMID- 11875734 TI - Alpha-foetoprotein in umbilical cord in relation to severe pre-eclampsia, birth weight and future breast cancer risk. AB - Women born after pre-eclamptic pregnancies have been reported to be at reduced risk of breast cancer as adults, because of reduced intrauterine oestrogen influence on breast tissue; high levels of alpha-foetoprotein (a glycoprotein with anti-oestrogenic properties), however, could also be important. In severe pre-eclampsia, placental function and foetal growth are reduced, and umbilical cord plasma levels of alpha-foetoprotein could reflect the underlying processes. Umbilical cord blood was collected in 12804 consecutive deliveries. Among 307 pregnancies with clinical pre-eclampsia, 66 singleton pregnancies were identified as clinically severe, and 610 singleton pregnancies were selected as controls. Oestradiol and alpha-foetoprotein were measured from umbilical plasma, and birth weight was standardized as the ratio between the observed and expected birth weight, adjusted for differences in gestation length and offspring sex. Cord plasma levels of alpha-foetoprotein were significantly higher in severe pre eclampsia than controls (P<0.01) after adjustment for gestational age and birth weight. For oestradiol, there was no difference in cord plasma levels between the severe pre-eclampsia group and controls, after adjustment for length of gestation and birth weight. These results suggest that an anti-oestrogenic effect associated with pre-eclampsia may be mediated through high levels of alpha foetoprotein rather than low levels of oestradiol. PMID- 11875735 TI - A case-control study of childhood leukaemia and paternal occupational contact level in rural Sweden. AB - In a national case-control study in Sweden, we investigated whether in rural areas (where susceptible individuals are more prevalent than in urban areas) leukaemia risk was higher among the young children of fathers with many work contacts, as the infective hypothesis has predicted. A total of 1935 cases diagnosed in 1958-1998 together with 7736 age-matched (within 1 year) population controls (of whom 970 and 3880 respectively were aged 0-4) were linked to paternal occupational details as recorded in the census closest to the year of birth. Applying the two classifications of occupational contact level used in a study of rural Scotland, the odds ratios for children aged 0-4 years in the highest contact category (which includes teachers) in the most rural Swedish counties were 3.47 (95% CI 1.54, 7.85) and 1.59 (1.07, 2.38) respectively, relative to the medium and low (reference) category; no such excess was found in urban or intermediate counties. There was also a significant positive trend at ages 0-4 in the rural counties across the three levels of increasing occupational contact (P for trend 0.02 and 0.03, respectively), but again not in the urban or intermediate counties. No such effect or trend was found at ages 5-14 in any of the three county groupings. The findings confirm those of a recent study in rural Scotland, and also suggest that unusual population mixing (as occurred in Scotland as a result of the North Sea oil industry) is not a necessary requirement for the effect, since comparable mixing has not been a feature of rural Sweden. PMID- 11875736 TI - Expression of different survivin variants in gastric carcinomas: first clues to a role of survivin-2B in tumour progression. AB - Survivin is a novel member of the inhibitor of apoptosis family and determines the susceptibility of tumour cells to pro-apoptotic stimuli. Recently, we identified two novel alternative splice variants of survivin, differing in their anti-apoptotic properties: whereas the anti-apoptotic potential of survivin DeltaEx3 is preserved, survivin-2B has lost its anti-apoptotic potential and may act as a naturally occurring antagonist of survivin. Because the in vivo expression of these alternative splice variants has not been explored so far, we analysed gastric carcinomas of different histological subtypes, grades and stages. Since no antibodies are currently available to determine the novel splice variants, quantitative reverse transcriptase polymerase chain reaction was performed, using RNA samples obtained from 30 different gastric carcinomas. Polymerase chain reactions products were quantified by densitometric evaluation. We found that all gastric carcinomas, irrespective of their histological types, grades or stages, express survivin-DeltaEx3, survivin-2B and survivin, the latter being the dominant transcript. Comparing the disease stages I+II with III+IV, expression of survivin and survivin-DeltaEx3 remained unchanged. In contrast, a significant (P=0.033) stage-dependent decrease in the expression of survivin-2B became evident. Our study demonstrates for the first time the expression of alternative splice variants in gastric carcinomas and provides a first clue to a role of survivin-2B in tumour progression. PMID- 11875737 TI - An analysis of p53, BAX and vascular endothelial growth factor expression in node positive rectal cancer. Relationships with tumour recurrence and event-free survival of patients treated with adjuvant chemoradiation. AB - Tumours of patients with node-positive rectal cancer were studied by immunohistochemistry for p53, BAX and vascular endothelial growth factor expressions. Results were correlated to the relapse rate, the pattern of relapse and the event-free survival after radical surgery and adjuvant chemoradiation. After a median follow-up of 60 months, 39 patients remained disease-free and 40 patients relapsed (18 local relapses and 22 distant metastases). The majority of disease-free patients showed p53 negative and vascular endothelial growth factor negative tumours. Local relapses occurred more frequently in patients with p53 overexpressing tumours (P<0.01), while distant metastases were in patients with vascular endothelial growth factor positive tumours (P<0.003). Patients with p53 negative or vascular endothelial growth factor negative tumours showed better event-free survival than patients with p53 positive or vascular endothelial growth factor positive tumours. BAX analysis did not show any association with patients' outcome and it was unrelated to the p53 status. Adjuvant treatment strategies for node-positive rectal cancer may be improved by identifying categories of high-risk patients. In this study, vascular endothelial growth factor and p53 expressions correlated with recurrent disease, pattern of relapse and poor event-free survival. PMID- 11875738 TI - p53 as a potential predictive factor of response to chemotherapy: feasibility of p53 assessment using a functional test in yeast from trucut biopsies in breast cancer patients. AB - Assessment of the predictive value of p53 requires the testing of large numbers of samples from patients enrolled in prospective phase III clinical trials. The goal of this study was to determine whether p53 status can be determined by p53 yeast functional assay using the limiting amounts of material that can typically be obtained in prospective phase III trials (particularly when chemotherapy is given before surgery). All patients presenting with a clinically palpable tumour which could be considered large enough to perform a trucut biopsy (> or =2 cm breast tumour) were eligible for this study. Two trucut biopsies and one incisional biopsy were performed on the surgical specimens (mastectomy or tumourectomy). Samples were snap frozen and cryostat sections were taken for histology and p53 testing. Thirty patients were included. Three samples out of 90 failed to give any p53 PCR products, probably because these samples contained almost entirely fibrous tissue. Of the 87 samples that could be tested, the incisional and trucut biopsies results were fully concordant in every case. p53 could be defined in 97% of patients by double trucut biopsy. Eight out of 30 tumours tested were mutant for p53 (27%). p53 status can be reliably determined by yeast assay from single frozen sections of trucut biopsies. Histological examination before p53 testing is essential to exclude cases where the p53 result may reflect only the status of the normal cells in the biopsy. PMID- 11875739 TI - Analysis of CHK2 in vulval neoplasia. AB - Structure and expression of the Rad53 homologue CHK2 were studied in vulval neoplasia. We identified the previously described silent polymorphism at codon 84 (A>G at nucleotide 252) in the germ-line of six out of 72, and somatic mutations in two out of 40 cases of vulval squamous cell carcinomas and none of 32 cases of vulval intraepithelial neoplasia. One mutation introduced a premature stop codon in the kinase domain of CHK2, whereas the second resulted in an amino acid substitution in the kinase domain. The two squamous cell carcinomas with mutations in CHK2 also expressed mutant p53. A CpG island was identified close to the putative CHK2 transcriptional start site, but methylation-specific PCR did not detect methylation in any of 40 vulval squamous cell carcinomas, irrespective of human papillomavirus or p53 status. Consistent with this observation, no cancer exhibited loss of CHK2 expression at mRNA or protein level. Taken together, these observations reveal that genetic but not epigenetic changes in CHK2 occur in a small proportion of vulval squamous cell carcinomas. PMID- 11875741 TI - Inhibitory effect on expression of angiogenic factors by antiangiogenic agents in renal cell carcinoma. AB - Since it has been widely recognised that renal cell carcinoma is refractory to standard therapies such as chemotherapy and radiotherapy, a new modality of treatment is needed. One of the potential alternative therapies for renal cell carcinoma may be inhibition of angiogenesis. In this study, we analysed the inhibitory effects of several potential agents on expression of angiogenic factors such as vascular endothelial growth factor and basic fibroblast growth factor, which are the main mediators in angiogenesis of renal cell carcinoma. We used medroxyprogesterone acetate, interferon-alpha, interferon-gamma, minocycline hydrochrolide and genistein, which are known to be antiangiogeneic. Northern blot analyses revealed that, among the five agents examined, genistein had a strong inhibitory effect on expression of vascular endothelial growth factor mRNA and basic fibroblast growth factor mRNA. Medroxyprogesterone acetate and interferon alpha did not significantly decrease the level of either vascular endothelial growth factor mRNA or basic fibroblast growth factor mRNA. Interferon-gamma and minocycline had mild inhibitory effects on vascular endothelial growth factor mRNA and basic fibroblast growth factor mRNA expression. Genistein also inhibited both vascular endothelial growth factor mRNA and basic fibroblast growth factor mRNA expression after treatment with epidermal growth factor and hypoxia. These findings suggest that one of the mechanisms of the inhibition of angiogenesis by genistein is suppression of the expression of the angiogenic factors vascular endothelial growth factor and basic fibroblast growth factor in renal cell carcinoma. PMID- 11875740 TI - Tamoxifen induction of angiogenic factor expression in endometrium. AB - Tamoxifen is the current therapy of choice for patients with oestrogen receptor positive breast cancer, and it is currently under evaluation as a prophylactic for women at high risk of developing the disease. However, tamoxifen is also known to induce proliferative changes in the endometrium increasing the risk of developing endometrial hyperplasia, polyps and carcinoma. Angiogenesis is an intimate part of this process. For this reason, we have examined the expression of several well characterized angiogenic factors, namely, acidic and basic fibroblast growth factor, thymidine phosphorylase, vascular endothelial growth factor and adrenomedullin in both normal and tamoxifen exposed pre- and postmenopausal endometrium. Vascular density and endothelial proliferation index were also quantified. We found increased expression of acidic and basic fibroblast growth factor and adrenomedullin after treatment with tamoxifen mainly in premenopausal tissue. Vascular density was significantly increased in pre- but not post-menopausal endometrium (P=0.0018) following tamoxifen treatment. These results support the notion that angiogenesis is integral to the response to tamoxifen exposure, and is a potential target with which to block these side effects of tamoxifen. PMID- 11875742 TI - The cancer preventative agent resveratrol is converted to the anticancer agent piceatannol by the cytochrome P450 enzyme CYP1B1. AB - Resveratrol is a cancer preventative agent that is found in red wine. Piceatannol is a closely related stilbene that has antileukaemic activity and is also a tyrosine kinase inhibitor. Piceatannol differs from resveratrol by having an additional aromatic hydroxy group. The enzyme CYP1B1 is overexpressed in a wide variety of human tumours and catalyses aromatic hydroxylation reactions. We report here that the cancer preventative agent resveratrol undergoes metabolism by the cytochrome P450 enzyme CYP1B1 to give a metabolite which has been identified as the known antileukaemic agent piceatannol. The metabolite was identified by high performance liquid chromatography analysis using fluorescence detection and the identity of the metabolite was further confirmed by derivatisation followed by gas chromatography-mass spectrometry studies using authentic piceatannol for comparison. This observation provides a novel explanation for the cancer preventative properties of resveratrol. It demonstrates that a natural dietary cancer preventative agent can be converted to a compound with known anticancer activity by an enzyme that is found in human tumours. Importantly this result gives insight into the functional role of CYP1B1 and provides evidence for the concept that CYP1B1 in tumours may be functioning as a growth suppressor enzyme. PMID- 11875743 TI - Thiram inhibits angiogenesis and slows the development of experimental tumours in mice. AB - Thiram-tetramethylthiuram disulphide--a chelator of heavy metals, inhibited DNA synthesis and induced apoptosis in cultured bovine capillary endothelial cells. Bovine capillary endothelial cells were 10-60-fold more sensitive to thiram than other cell types. These effects were prevented by addition of antioxidants, indicating involvement of reactive oxygen species. Exogenously added Cu2+ impeded specifically and almost completely the inhibitory effect of thiram for bovine capillary endothelial cells. Moreover, thiram had markedly inhibited human recombinant Cu/Zn superoxide dismutase enzymatic activity (85%) in vitro. Moreover, PC12-SOD cells with elevated Cu/Zn superoxide dismutase were less sensitive to thiram treatment than control cells. These data indicate that the effects of thiram are mediated by inhibition of Cu/Zn superoxide dismutase activity. Oral administration of thiram (13-30 microg mouse(-1)), inhibited angiogenesis in CD1 nude mice. Tumour development is known to largely depend on angiogenesis. We found that oral administration of thiram (30 microg) to mice caused significant inhibition of C6 glioma tumour development (60%) and marked reduction (by 3-5-fold) in metastatic growth of Lewis lung carcinoma. The data establish thiram as a potential inhibitor of angiogenesis and raise the possibility for its use as therapy in pathologies in which neovascularisation is involved, including neoplasia. PMID- 11875744 TI - Inhibition of the alpha-nu integrins with a cyclic RGD peptide impairs angiogenesis, growth and metastasis of solid tumours in vivo. AB - Anti-angiogenetic cancer therapy is a potential new form for treatment of solid tumours. The alpha(v)-integrins (alpha(v)beta3, alpha(v)beta5) mediate the contact of activated endothelial cells to proteins of the extracellular matrix during tumour angiogenesis as a prerequisite for survival of endothelial cells. The aim of this study was to investigate the effects of application of a methylated cyclic RGD-peptide as an alpha(v)-integrin antagonist on angiogenesis, microcirculation, growth and metastasis formation of a solid tumour in vivo. Experiments were performed in the dorsal skinfold preparation of Syrian Golden hamsters bearing the amelanotic hamster melanoma A-Mel-3. Animals were injected intraperitoneally with a methylated cyclic RGD-peptide every 12 h, the control group received an inactive peptide. Microcirculatory parameters of tumour angiogenesis including functional vessel density, red blood cell velocity, vessel diameter and leucocyte-endothelium interaction were analysed using intravital microscopy. In an additional study the effects on growth and metastasis of subcutaneous A-Mel-3 were quantified. Functional vessel density was markedly reduced on day 3 in treated animals compared to controls (37.2 +/- 12.1 vs 105.2 +/- 11.2 cm(-1); mean +/- s.e.m.; P<0.05) and increased subsequently in both groups. Red blood cell velocity at day 3 was below values of controls (0.026 +/- 0.01 vs 0.12 +/- 0.03 mm x s(-1); P<0.05). No differences were observed in vessel diameters and leucocyte-endothelium interaction was almost absent in both groups. Furthermore, growth and metastasis of subcutaneous tumours after administration of the cyclic RGD-peptide was significantly delayed in comparison to controls (P<0.05). Inhibition of alpha(v)-integrins by a cyclic RGD-peptide resulted in significant reduction of functional vessel density, retardation of tumour growth and metastasis in vivo. Taken together, these results implicate RGD-peptides as agents which have anti-tumour and anti-metastatic activity in vivo. PMID- 11875745 TI - Murine 5T multiple myeloma cells induce angiogenesis in vitro and in vivo. AB - Multiple myeloma is a B cell malignancy. Recently, it has been demonstrated that bone marrow samples of patients with multiple myeloma display an enhanced angiogenesis. The mechanisms involved seem to be multiple and complex. We here demonstrate that the murine 5T multiple myeloma models are able to induce angiogenesis in vitro by using a rat aortic ring assay and in vivo by determining the microvessel density. The rat aortic rings cultured in 5T multiple myeloma conditioned medium exhibit a higher number of longer and more branched microvessels than the rings cultured in control medium. In bone marrow samples from 5T multiple myeloma diseased mice, a statistically significant increase of the microvessel density was observed when compared to bone marrow samples from age-matched controls. The angiogenic phenotype of both 5T multiple myeloma cells could be related, at least in part, to their capacity to produce vascular endothelial growth factor. These data clearly demonstrate that the 5T multiple myeloma models are good models to study angiogenesis in multiple myeloma and will allow to unravel the mechanisms of neovascularisation, as well as to test new putative inhibitors of angiogenesis. PMID- 11875746 TI - Anti-angiogenic effects of the thienopyridine SR 25989 in vitro and in vivo in a murine pulmonary metastasis model. AB - Neovascularisation is a key step in tumour growth and establishment of distant metastases. We have recently demonstrated that the thienopyridine SR 25989 an enantiomer of the anti-aggregant clopidogrel (Plavix) lacking anti-aggregant activity, inhibits endothelial cell proliferation in vitro by increasing the expression of endogenous thrombospondin-1, a natural potent inhibitor of angiogenesis. The anti-angiogenic effect of SR 25989 was further assessed in vitro in a quantitative assay of angiogenesis comprising a fragment of rat aorta embedded in a fibrin gel and in vivo in a pulmonary metastatic model using C57BL/6 mice inoculated in the foot pad with the highly metastatic melanoma cell line B16 F10. SR 25989 induced a dose dependent inhibition of spontaneous microvessel development in vitro reaching half maximal inhibition at around less than 50 microM and caused platelet derived growth factor induced angiogenesis to regress as a function of thienopyridine concentration. In vivo, SR 25989 did not alter significantly the growth rate of the primary tumour in the foot pad and did not inhibit development of inguinal nodes which appeared after amputation. However, the number and size of lung metastases were reduced in treated animals when examined at the time of sacrifice. In addition, the few metastases over 1 mm3 did not show any neovascularisation, as confirmed by negative von Willebrand immunostaining and in contrast to intense vascularisation seen in metastases developed by control mice. These results confirm that SR 25989 possesses potent anti-angiogenic properties and is able to inhibit metastatic dissemination and growth. The lack of effect on the primary tumour and inguinal nodes illustrates the complexity of the mechanisms involved in tumoural neo-angiogenesis and points out the possibility for distinct processes leading to neovascularisation in primary tumour as opposed to metastases. PMID- 11875747 TI - A fully human anti-Ep-CAM scFv-beta-glucuronidase fusion protein for selective chemotherapy with a glucuronide prodrug. AB - Monoclonal antibodies against tumour-associated antigens could be useful to deliver enzymes selectively to the site of a tumour for activation of a non-toxic prodrug. A completely human fusion protein may be advantageous for repeated administration, as host immune responses may be avoided. We have constructed a fusion protein consisting of a human single chain Fv antibody, C28, against the epithelial cell adhesion molecule and the human enzyme beta-glucuronidase. The sequences encoding C28 and human enzyme beta-glucuronidase were joined by a sequence encoding a flexible linker, and were preceded by the IgGkappa signal sequence for secretion of the fusion protein. A CHO cell line was engineered to secrete C28-beta-glucuronidase fusion protein. Antibody specificity and enzyme activity were retained in the secreted fusion protein that had an apparent molecular mass of 100 kDa under denaturing conditions. The fusion protein was able to convert a non-toxic prodrug of doxorubicin, N-[4-doxorubicin-N carbonyl(oxymethyl)phenyl]-O-beta-glucuronyl carbamate to doxorubicin, resulting in cytotoxicity. A bystander effect was demonstrated, as doxorubicin was detected in all cells after N-[4-doxorubicin-N-carbonyl(oxymethyl)phenyl]-O-beta glucuronyl carbamate administration when only 10% of the cells expressed the fusion protein. This is the first fully human and functional fusion protein consisting of an scFv against epithelial cell adhesion molecule and human enzyme beta-glucuronidase for future use in tumour-specific activation of a non-toxic glucuronide prodrug. PMID- 11875748 TI - Sequence-dependent effects of ZD1839 ('Iressa') in combination with cytotoxic treatment in human head and neck cancer. AB - Elevated levels of epidermal growth factor receptor in head and neck cancer have been extensively reported, and are correlated with poor prognosis. The combination of cisplatin and 5-fluorouracil is a standard treatment regimen for head and neck cancer, with radiation representing another therapeutic option. Six head and neck cancer cell lines were used to study the cytotoxic effects of combining ZD1839 ('Iressa'), a new selective epidermal growth factor receptor tyrosine kinase inhibitor, and radiation. Two of the cell lines were also used to study the combination of ZD1839 and cisplatin/5-fluorouracil. Cytotoxic effects were assessed by the MTT test. The results indicated that ZD1839 applied before radiation gave the best effects (P=0.002); an effect that was strongest in those p53-mutated cell lines that express the highest epidermal growth factor receptor levels. The effects of ZD1839 with cisplatin and/or 5-fluorouracil were sequence dependent (P<0.003), with the best results achieved when ZD1839 was applied first. For the triple combinations, ZD1839 applied before cisplatin and 5 fluorouracil resulted in a slight synergistic effect (P=0.03), although the effect was greater when ZD1839 was applied both before and during cytotoxic drug exposure. In conclusion, ZD1839 applied before radiation and before and/or during cisplatin/5-fluorouracil may improve the efficacy of treatment for head and neck cancer. PMID- 11875749 TI - Superantigen reactive Vbeta6+ T cells induce perforin/granzyme B mediated caspase independent apoptosis in tumour cells. AB - The endogenous viral superantigen 7 in DBA/2 mice serves as a target antigen on syngeneic ESb-MP lymphoma cells for allogeneic graft-vs-leukaemia reactive cells. Allogeneic viral superantigen 7 reactive Vbeta6+ T cells are able to transfer graft-vs-leukaemia reactivity and to kill specifically viral superantigen 7+ ESb MP tumour cells in vitro. Here we elucidate the mechanism of this superantigen specific cell lysis. Already 10 min after co-incubation with in vitro stimulated Vbeta6+ T cells, viral superantigen 7+ ESb-MP tumour cells show an apoptotic phenotype (Annexin V-positivity, DNA-fragmentation). This extremely rapid type of cell death is not mediated by the death inducing ligands CD95L, TRAIL and TNF but by perforin and granzyme B. Surprisingly, neither mitochondria nor any of the known caspases appear to be involved in this type of tumour cell killing. In contrast, nitric oxide, released by activated macrophages and endothelial cells, induces in the same tumour cells another type of apoptosis which is much slower and involves mitochondria and caspase activation. A synergistic effect between the two different effector mechanisms of superantigen reactive donor cytotoxic T lymphocytes and nitric oxide releasing host macrophages and endothelial cells might explain the effective immune rejection of even advanced metastasised cancer in this graft-vs-leukaemia animal model. PMID- 11875751 TI - Transplantation of islets of Langerhans: new developments. AB - The clinical results recently reported by the Edmonton group in recipients of allogeneic islet grafts, all of whom achieved at least temporary insulin independence, has rekindled interest in transplantation of islets of Langerhans as a means to cure diabetes. Long-term islet graft survival has been achieved in a non-human primate pre-clinical model with a protocol of T-cell signaling blockade using a new monoclonal antibody. Islet xenotransplantation (namely the use of animal islets, with the aim of transplanting them into humans), or stem cell technology (the controlled differentiation of stem cells to obtain specialised cells for the treatment of diabetes) are other procedures currently being evaluated in animal models. The recent clinical success suggests that, in the near future, diabetes might be treated by islet transplantation early in the clinical course of the disease before the development of complications, and without the risks associated with conventional immunosuppression. PMID- 11875750 TI - ALA and ALA hexyl ester in free and liposomal formulations for the photosensitisation of tumour organ cultures. AB - In spite of the wide range of tumours successfully treated with 5-aminolevulinic acid mediated photodynamic therapy, the fact that 5-aminolevulinic acid has low lipid solubility, limits its clinical application. More lipophilic 5 aminolevulinic acid prodrugs and the use of liposomal carriers are two approaches aimed at improving 5-aminolevulinic acid transmembrane access. In this study we used both 5-aminolevulinic acid and its hexyl ester in their free and encapsulated formulations to compare their corresponding endogenous synthesis of porphyrins. Employing murine tumour cultures, we found that neither the use of hexyl ester nor the entrapment of either 5-aminolevulinic acid or hexyl ester into liposomes increase the rate of tumour porphyrin synthesis. By light and electronic microscopy it was demonstrated that exposure of tumour explants to either free or liposomal 5-aminolevulinic acid and subsequent illumination induces the same type of subcellular damage. Mitochondria, endoplasmic reticulum and plasma membrane are the structures mostly injured in the early steps of photodynamic treatment. In a later stage, cytoplasmic and nuclear disintegration are observed. By electronic microscopy the involvement of the endocytic pathway in the incorporation of liposomal 5-aminolevulinic acid into the cells was shown. PMID- 11875753 TI - A year's review of bacterial pneumonia at the central hospital of Lucerne, Switzerland. AB - Community acquired pneumonia (CAP) remains an important cause of substantial morbidity and mortality in inhospital patients. We conducted a retrospective study of all patients hospitalised at our hospital with the diagnosis of bacterial pneumonia according to ICD-10 within one year. Of 360 identified charts, 335 met the requirements and were reviewed regarding risk factors, diagnosis, treatment, and overall mortality. The typical patient hospitalised with pneumonia was elderly (mean 68 years), male (60%), and suffered from comorbidities or predisposing factors (96.4%). A total of 72.8% of pneumonias were localized in the inferior lobes, and 21.1% had bilateral infiltrates. Etiologic agents were searched for in 297/335 patients (87.5%) and were found positive in 33.4%: of 169 blood cultures 9.5% were positive, of 150 sputum cultures taken 46.6% were positive, of 17 serologies taken 58.8% were positive, and of 9 pleural effusions analysed 22.2% were positive. Encapsulated bacteria were the most common found bacterial etiologies, namely Streptococcus pneumoniae (S. pneumoniae) in 30.9% of patients with known bacterial etiology, Haemophilus in 24.7%, and Klebsiella in 12.4%. Methicillin-resistant S. aureus was not found. The three most commonly used antibiotics were amoxicillin/clavulanic acid used in 77.3% of patients, clarithromycin (41.2%), and ceftriaxone (16.6%). Mean duration of treatment was 12.1 days. 245/335 (73.1%) patients had a favourable outcome, 16.7% (56/335) of patients had a protracted illness with delayed resolution (i.e. prolonged hospital stay, need for intensive care, intubation or several of these complications). Overall mortality in our unit was 8.6%. PMID- 11875752 TI - Intensive care unit admission in patients with haematological disease: incidence, outcome and prognostic factors. AB - OBJECTIVES: To examine incidence and outcome of intensive care unit (ICU) admission in patients with haematological malignancy and analyse prognostic factors associated with outcome. DESIGN: Retrospective cohort study in an intensive care unit of a tertiary referral center. PATIENTS: 78 patients with severe haematological malignancy were admitted 97 times between 1990-97 to the medical ICU for septic shock (18), respiratory failure (30), postoperative monitoring (19), cardiovascular (10) and central nervous complications (8) or for other reasons (12). Median age was 43 (4-73) years, average duration of ICU stay was 4 (1-43) days. Forty-two patients required mechanical ventilation, 46 vasopressors and 8 haemodialysis. RESULTS: Rates of ICU admission differed by treatment of the underlying disease. There were 18, 10 and 27 ICU admissions per 100 treatments in patients undergoing chemotherapy for acute leukaemia, autologous and allogeneic stem cell transplantation (p <0.005) respectively. Thirty-two of 78 patients died within 60 days of ICU admission. Organ failure, i.e. cardiovascular failure requiring vasopressors, respiratory failure requiring mechanical ventilation and renal failure, requiring haemodialysis, was most significantly associated with outcome. Mortality by day 60 after admission was 16%, 36%, 64%, and 83% (p <0.0002) for patients without organ failure, and for patients with 1, 2 or 3 failing organs. In a multivariate logistical regression model, only the organ failure score (p <0.0005) and evidence of liver damage, defined as ASAT or ALAT >100 IU/ L (p <0.007), but not age, sex, primary disease and treatment of the underlying disease predicted outcome. CONCLUSION: Multi organ failure and evidence of liver damage but no other patient, disease, or treatment related factor predict outcome in patients with haematological disease admitted to the ICU. PMID- 11875754 TI - Repetitive intraoperative use of recombinant hirudin (Lepirudin) in peripheral vascular surgery with HITT. AB - Heparin induced thrombocytopenia with thrombosis (HITT) is a rare but dangerous complication related to the application of unfractionated heparin or low molecular weight heparin. Due to an antibody dependent in vivo platelet activation, severe thromboembolic episodes may occur. We present the case of a patient with HITT following implantation of an aortobifemoral graft secondary to bilateral common iliac artery stenoses. An arterial clot developed and led to a partial occlusion of the graft to the right external iliac artery. Heparin was replaced by Lepirudin, a recombinant hirudin. A bolus of 0.4 mg/kg body weight was given, thereafter 0.15 mg/kg body weight per hour was administered continuously i.v. to maintain the aPTT 2- to 2.5-fold above the baseline value. The platelet count (minimum 47 G/l) normalised within two days. During thrombectomy of the right common femoral artery we used Lepirudin intraoperatively (bolus injection of 0.2 mg/kg body weight) to prevent any further platelet and coagulation activation during the clamping phase. Six months later the patient underwent two further bypass operations due to severe stenoses of both superficial femoral arteries. Due to the high risk of thromboembolism if HITT recurred, a bolus of 0.2 mg/kg body weight of Lepirudin was given during each intervention. No bleeding complications occurred. In addition Lepirudin appeared to decrease platelet consumption in the absence of active thrombosis. Direct thrombin inhibitors such as Lepirudin possess no heparin-like immunological properties and seem to have become the therapeutic "gold-standard" in patients with HITT. Our experience suggests that the repetitive intraoperative use of Lepirudin is safe and effective. PMID- 11875755 TI - CARD15/NOD2 mutational analysis and genotype-phenotype correlation in 612 patients with inflammatory bowel disease. AB - CARD15/NOD2 encodes a protein involved in bacterial recognition by monocytes. Mutations in CARD15 have recently been found in patients with Crohn disease (CD), a chronic inflammatory condition of the digestive tract. Here, we report the mutational analyses of CARD15 in 453 patients with CD, including 166 sporadic and 287 familial cases, 159 patients with ulcerative colitis (UC), and 103 healthy control subjects. Of 67 sequence variations identified, 9 had an allele frequency >5% in patients with CD. Six of them were considered to be polymorphisms, and three (R702W, G908R, and 1007fs) were confirmed to be independently associated with susceptibility to CD. Also considered as potential disease-causing mutations (DCMs) were 27 rare additional mutations. The three main variants (R702W, G908R, and 1007fs) represented 32%, 18%, and 31%, respectively, of the total CD mutations, whereas the total of the 27 rare mutations represented 19% of DCMs. Altogether, 93% of the mutations were located in the distal third of the gene. No mutations were found to be associated with UC. In contrast, 50% of patients with CD carried at least one DCM, including 17% who had a double mutation. This observation confirmed the gene-dosage effect in CD. The patients with double-dose mutations were characterized by a younger age at onset (16.9 years vs. 19.8 years; P=.01), a more frequent stricturing phenotype (53% vs. 28%; P=.00003; odds ratio 2.92), and a less frequent colonic involvement (43% vs. 62%; P=.003; odds ratio 0.44) than were seen in those patients who had no mutation. The severity of the disease and extraintestinal manifestations were not different for any of the CARD15 genotypes. The proportion of familial and sporadic cases and the proportion of patients with smoking habits were similar in the groups of patients with CD with or without mutation. These findings provide tools for a DNA-based test of susceptibility and for genetic counseling in inflammatory bowel disease. PMID- 11875756 TI - Phenotypic homogeneity provides increased support for linkage on chromosome 2 in autistic disorder. AB - Autistic disorder (AutD) is a neurodevelopmental disorder characterized by significant disturbances in social, communicative, and behavioral functioning. A two-stage genomic screen analysis of 99 families with AutD revealed suggestive evidence for linkage to chromosome 2q (D2S116 nonparametric sib-pair LOD score [MLS] 1.12 at 198 cM). In addition, analysis of linkage disequilibrium for D2S116 showed an allele-specific P value of <.01. Recently, linkage to the same region of 2q was reported in an independent genome screen. This evidence for linkage increased when analysis was restricted to the subset of patients with AutD who had delayed onset (>36 mo) of phrase speech (PSD). We similarly classified our data set of 82 sib pairs with AutD, identifying 45 families with AutD and PSD. Analysis of this PSD subset increased our support for linkage to 2q (MLS 2.86 and HLOD 2.12 for marker D2S116). These data support evidence for a gene on chromosome 2 contributing to risk of AutD, and they suggest that phenotypic homogeneity increases the power to find susceptibility genes for AutD. PMID- 11875757 TI - The evolutionary origin of human subtelomeric homologies--or where the ends begin. AB - The subtelomeric regions of human chromosomes are comprised of sequence homologies shared between distinct subsets of chromosomes. In the course of developing a set of unique human telomere clones, we identified many clones containing such shared homologies, characterized by the presence of cross hybridization signals on one or more telomeres in a fluorescence in situ hybridization (FISH) assay. We studied the evolutionary origin of seven subtelomeric clones by performing comparative FISH analysis on a primate panel that included great apes and Old World monkeys. All clones tested showed a single hybridization site in Old World monkeys that corresponded to one of the orthologous human sites, thus indicating the ancestral origin. The timing of the duplication events varied among the subtelomeric regions, from approximately 5 to approximately 25 million years ago. To examine the origin of and mechanism for one of these subtelomeric duplications, we compared the sequence derived from human 2q13--an ancestral fusion site of two great ape telomeric regions--with its paralogous subtelomeric sequences at 9p and 22q. These paralogous regions share large continuous homologies and contain three genes: RABL2B, forkhead box D4, and COBW-like. Our results provide further evidence for subtelomeric-mediated genomic duplication and demonstrate that these segmental duplications are most likely the result of ancestral unbalanced translocations that have been fixed in the genome during recent primate evolution. PMID- 11875760 TI - Comprehensive critical care--foundation for the future? A selection of abstracts from 1 September and 3 November 2000. PMID- 11875758 TI - Age at onset in two common neurodegenerative diseases is genetically controlled. AB - To identify genes influencing age at onset (AAO) in two common neurodegenerative diseases, a genomic screen was performed for AAO in families with Alzheimer disease (AD; n=449) and Parkinson disease (PD; n=174). Heritabilities between 40% -60% were found in both the AD and PD data sets. For PD, significant evidence for linkage to AAO was found on chromosome 1p (LOD = 3.41). For AD, the AAO effect of APOE (LOD = 3.28) was confirmed. In addition, evidence for AAO linkage on chromosomes 6 and 10 was identified independently in both the AD and PD data sets. Subsequent unified analyses of these regions identified a single peak on chromosome 10q between D10S1239 and D10S1237, with a maximum LOD score of 2.62. These data suggest that a common gene affects AAO in these two common complex neurodegenerative diseases. PMID- 11875761 TI - Distinct histopathological patterns in single lesion leprosy patients treated with single dose therapy (ROM) in the Brazilian Multicentric Study. AB - This paper aims to describe the histomorphologic features of skin biopsies of single lesion leprosy patients recruited at outpatient clinics in four Brazilian states in the Northeast (Amazonas and Rondonia), Southeast (Rio de Janeiro) and Center-West (Goias) between October 1997 and December 1998. Patients clinically diagnosed as single skin lesion paucibacillary (SSL-PB) leprosy had a standard 4 mm punch biopsy taken from the lesion before rifampin, ofloxacin, minocycline (ROM) therapy. The features of the cellular inflammatory infiltrates, the presence of nerve involvement and acid-fast bacilli (AFB) were used to categorize SSL-PB biopsies into different histopathological groups. Two-hundred-seventy eight (93.0%) out of 299 patients had a skin biopsy available. Seven single lesion patients were diagnosed as BL or LL leprosy types (MB) by the histopathological exams and 12 cases were excluded due to other skin diseases. Therefore, 259 patients had skin lesions with histomorphological features compatible with PB leprosy categorized as follows: 33.6% (N = 87) of the biopsies represented well-circumscribed epithelioid cell granuloma (Group 1); 21.6% (N = 56) less-circumscribed epithelioid cell granuloma (Group 2); 12.0% (N = 31) were described as mononuclear inflammatory infiltrate permeated with epithelioid cells (Group 3), and 29.7% (N = 77) had perivascular/periadnexal mononuclear inflammatory infiltrate (Group 4). Minimal/no morphological alteration in the skin was detected in only 8 (3.1%) SSL-PB patients categorized as Group 5, who were considered to have leprosy by clinical parameters. SSL-PB leprosy patients recruited in a multicentric study presented histomorphology readings comprising the whole PB leprosy spectrum but also a few MB cases. These results indicate heterogeneity among SSL-PB patients, with a predominance of well-circumscribed and less-circumscribed epithelioid cell granulomas (Groups 1 and 2) in the sites studied and the heterogeneity of local cellular immune response. PMID- 11875762 TI - Impact of combined Mycobacterium w vaccine and 1 year of MDT on multibacillary leprosy patients. AB - A total of 20 bacteriologically positive multibacillary (MB) leprosy patients older than 18 years of age with a bacterial index (BI) of 2+ or greater were given standard World Health Organization multiple drug therapy (MDT-MB) for 12 consecutive months plus four intradermal doses of Mycobacterium w vaccine at 3 monthly intervals (Study group). Twenty age-matched MB patients were given WHO/MDT alone (Control group). The patients of both groups were followed up for 1 year. Improvements in the patients were periodically monitored by clinical (Ramu's score), bacteriological (SSS), histopathological (skin biopsy) and immunological (lepromin conversion) parameters. Study group patients showed more significant improvements in all parameters except for lepromin conversion compared to patients in the Control group. The incidence of type 1 reaction was more in the Study group (30% vs 10%), while the incidence of type 2 reaction was more in the Control group (25% vs 15%). Neuritis associated with reactions was seen more often in the Control group compared to the Study group (20% vs 10%). The addition of Mycobacterium w vaccine as an adjunct to the 1-year WHO/MDT regimen appears to be significantly more beneficial in MB leprosy patients with a high initial BI compared to WHO/MDT given alone. Studies on larger numbers of patients with extended follow up will be in order. PMID- 11875759 TI - Protean PTEN: form and function. AB - Germline mutations distributed across the PTEN tumor-suppressor gene have been found to result in a wide spectrum of phenotypic features. Originally shown to be a major susceptibility gene for both Cowden syndrome (CS), which is characterized by multiple hamartomas and an increased risk of breast, thyroid, and endometrial cancers, and Bannayan-Riley-Ruvalcaba syndrome, which is characterized by lipomatosis, macrocephaly, and speckled penis, the PTEN hamartoma tumor syndrome spectrum has broadened to include Proteus syndrome and Proteus-like syndromes. Exon 5, which encodes the core motif, is a hotspot for mutations likely due to the biology of the protein. PTEN is a major lipid 3-phosphatase, which signals down the PI3 kinase/AKT pro-apoptotic pathway. Furthermore, PTEN is a protein phosphatase, with the ability to dephosphorylate both serine and threonine residues. The protein-phosphatase activity has also been shown to regulate various cell-survival pathways, such as the mitogen-activated kinase (MAPK) pathway. Although it is well established that PTEN's lipid-phosphatase activity, via the PI3K/AKT pathway, mediates growth suppression, there is accumulating evidence that the protein-phosphatase/MAPK pathway is equally important in the mediation of growth arrest and other crucial cellular functions. PMID- 11875763 TI - Immunological profile of treated lepromatous leprosy patients. AB - The immune responses of 19 treated lepromatous patients who had remained smear negative for a long period were assessed for specific cell-mediated immunity (CMI), anti-Mycobacterium leprae antibodies and cytokine release in response to challenge with M. leprae soluble antigen (MLSA). All of these patients remained anergic to Mitsuda lepromin. Lymphoproliferation in response to M. leprae antigen was noted in only two patients. Significant reduction in the phenolic glycolipid I (PGL-I) antibody response in treated patients with no difference in the M. leprae 35-kDa antibody response was observed when these responses were compared with those of active lepromatous patients. More treated patients produced interleukin-2 (IL-2) and interferon gamma (IFN-gamma) than did active patients. On the other hand, fewer treated patients produced IL-10 than did active patients. These limited findings suggest that the host immune response makes an attempt toward upregulation of CMI in some treated LL/BL patients. PMID- 11875764 TI - Cytokine profiles in paraffin-embedded biopsy samples of lepromatous leprosy patients: semi-quantitative measure of cytokine mRNA using RT-PCR. AB - A reproducible technique for fixation of tissue, RNA extraction and reverse transcription polymerase chain reaction (RT-PCR) analysis from paraffin-embedded leprosy biopsies, has been developed and used to study the mRNA profiles. This approach is valuable in retrospective analysis of gene expression, and the handling of infectious biopsy material is also minimized. Among the methods of RNA extraction compared, the most efficient method was found to be incubation of the tissue sections in digestion buffer followed by extraction with Trizol. The experimental conditions were optimized for first strand cDNA synthesis and PCR, and used to measure the quantity of cytokine transcripts in biopsy materials. Interleukin-10 (IL-10) mRNA was detectable in all cases examined, which correlates well with other earlier reports using frozen tissues. However, IL-5 transcripts were present in 60% of the biopsies, unlike the earlier reports which showed IL-5 mRNA in all LL cases. Transforming growth factor-beta (TGF-beta) mRNA was detected in 80% of the biopsies, and this confirmed earlier immuno cytochemical data which showed TGF-beta protein in all cases. Tumor necrosis factor-alpha mRNA was found in about 60% of the biopsies; whereas interferon gamma mRNA was detected in 30% of the LL cases. In conclusion, the results obtained in our study confirm and extend earlier observations which examined cytokines in peripheral blood cells and dermal lesions of leprosy. The simplicity of this method would allow the examination of a large number of samples across the spectrum of leprosy. PMID- 11875765 TI - Pathogenesis of dry eye in leprosy and tear functions. AB - Seventy-five leprosy patients and an equal number of age- and sex-matched controls were examined for tear functions, using Schirmer's test and tear break up time (BUT). There was no statistically significant difference in the Schirmer's test, but the tear BUT showed a statistically significant lower value of < 10 seconds in multibacillary patients compared to paucibacillary patients. Leprosy patients with lagophthalmos and decreased corneal sensation showed a lower value of tear BUT which was also statistically significant. This study shows that even though the quantity of tears is not affected, proper and prolonged wetting of the cornea is deficient in many leprosy patients. PMID- 11875766 TI - MRI in clinically asymptomatic neuropathic leprosy feet: a baseline study. AB - This study was undertaken to analyze the magnetic resonance imaging (MRI) findings in the clinically asymptomatic neuropathic feet of leprosy patients. Since in the literature no MRI data are available concerning the asymptomatic neuropathic foot in leprosy, the interpretation of MRI examinations in clinically suspected neuropathic feet in leprosy is difficult. We examined 10 adult leprosy patients with clinically asymptomatic neuropathic feet. Inclusion criteria were a normal or near normal neuropathic foot, without signs of inflammation. All patients underwent an MRI protocol with the inclusion of two-point Dixon chemical shift imaging as fat suppression sequence. We found MRI changes in almost all patients. The most striking were the changes located in the region of the first metacarpophalangeal (MTP) joint. These changes ranged from degradation and interruption of the subcutaneous fat to effusion/synovitis in the first MTP joint. This study reveals significant MRI changes in clinically asymptomatic neuropathic feet in patients with leprosy. These changes may relate to the development of ulcerations. MRI may play an important role in detecting feet at risk and may influence clinical decision making. PMID- 11875767 TI - Pentoxifylline downregulates nitric oxide and tumor necrosis factor-alpha induced by mycobacterial lipoarabinomannan in a macrophage cell line. AB - Pentoxifylline (PTX), a phosphodiesterase inhibitor, is known to downregulate tumor necrosis factor-alpha (TNF-alpha) secretion induced by lipopolysacchride (LPS) and gamma interferon (IFN-gamma). We have had limited success in treating leprosy reactions, including erythema nodosum leprosum (ENL), in which TNF-alpha has been identified as a major proinflammatory cytokine. PTX inhibited production of NO (IC50 approximately equal to 1.0 mg/ml) and TNF-alpha (IC50 approximately equal to 0.05 mg/ml) in a dose-dependent fashion. As little as 0.5 mg/ml of PTX decreased NO production and 0.01 mg/ml of PTX inhibited TNF-alpha production. Western blot analyses demonstrated that iNOS was suppressed by PTX. Northern blot analyses showed significant reduction of TNF-alpha mRNA. We conclude that PTX is an effective inhibitor of lipoarabinomannan (LAM)-induced TNF-alpha production at both the product and transcriptional levels in our macrophage cell line. PTX also showed moderate inhibition of NO at the product level as well as translation of iNOS. PMID- 11875768 TI - Some epidemiological observations on leprosy in India. AB - This population sample survey conducted in rural and urban areas of the Agra District in India showed an active leprosy caseload of 60.1/10,000 in the rural and 39.1/10,000 in the urban areas against a targeted prevalence of < 1/10,000. The disease appeared to be widespread since almost 65% of the villages or urban pockets surveyed had at least one prevalent case of leprosy. Significantly larger numbers of leprosy patients were found among males, agricultural/manual workers, persons with no formal schooling, individuals living in unkept households with dirty surroundings, and among those living in dark and poorly ventilated houses. The epidemiological significance of this study reveals the endemic nature of leprosy in Agra and suggests the need to intensify and widen case-detection activities to achieve leprosy control. PMID- 11875769 TI - Acceptance of WHO/MDT over the last 20 years. PMID- 11875770 TI - Dapsone-induced methemoglobinemia in leprosy patients. PMID- 11875771 TI - Tumor necrosis factor (TNF) production in leprosy patients. PMID- 11875772 TI - Leprosy and neurofibromatosis. 2: What is common? PMID- 11875773 TI - Report on the 1st annual National Science and Policy Research Consensus Conference on Health and Climate Change--Ottawa, 13 to 15 March, 2001, and implications for infectious diseases. PMID- 11875774 TI - An advisory committee statement (ACS). National Advisory Committee on Immunization (NACI). NACI update to statement on varicella vaccine. PMID- 11875775 TI - [Mannose receptor]. AB - The mannose receptor (MR) is a calcium-dependent C-type lectin that is expressed on some macrophages, immature dendritic cells, endothelial, and epithelial cells. The most important feature of MR is the capability to distinguish self from non self molecules through the interaction of multiple CRDs. The purpose of the short review is to summarise the structural and functional properties of MR. PMID- 11875776 TI - [The role of components of peritoneal fluid in the immunopathogenesis of endometriosis]. AB - Endometriosis is characterised by the presence of abnormally located tissue resembling the endometrium with glands and stroma. The abnormal endometrium of women affected by endometriosis could be able to protect itself from harmful effects of immune cells by expressing specific antigens, by harbouring a different population and by synthesizing and secreting immunosuppressive factors. This disturbance may be regulated by factors in peritoneal fluid which is a specific microenvironment. PMID- 11875777 TI - [NK cells. II. Their phenotypic and functional heterogeneity]. AB - This article reviews of recent data on the phenotypic markers and receptors of human and animal NK cells. It informs of the NK cell phenotypic heterogeneity and the functional dependence of NK cells on the modulatory action of their activating and inhibitory receptors. Moreover, it describes the NK killing mechanisms. PMID- 11875778 TI - [Participation of final products of lipid peroxidation in the anticancer mechanism of ionizing radiation and radiomimetic cytostatics]. AB - This review reports the evidence for the participation of final products of lipid peroxidation in the anticancer mechanism of ionising radiation and radiomimetic cytostatics. Processes of lipid peroxidation occur endogenously in response to oxidative stress and great diversity of reactive metabolites is formed. However, direct observation of radical reaction in pathophysiology of cells, tissues and organs is limited technically. Most investigations focused on the indirect assessment of their final products, aldehydes. The peroxidative breakdown of polyunsaturated fatty acids is believed to be involved in the regulation of cell division, and antitumor effect through biochemical and genetic processes. PMID- 11875780 TI - [The role of arginine vasopressin (AVP) in pain transmission and perception]. AB - In mammalian organisms were found composed systems controlling the transmission and perception of nociceptive impulses. The present review focuses on clinical and laboratory studies that are aimed at identifying the role of AVP, known antidiuretic hormone in the mechanism of pain phenomenon. PMID- 11875779 TI - [Influence of abnormalities of TP53 and ATM genes for course of B-cell chronic lymphocytic leukemia]. AB - Uncontrolled cell proliferation is the hallmark of cancer, and tumour cells have typically acquired damage to genes that directly regulate their cell cycles. Genes that link checkpoint controls to cancer and other related genetic disease include the TP53 and ATM genes. The abnormalities of these genes are clearly associated with particular forms of the B-cell chronic lymphocytic leukemia and poor prognosis. PMID- 11875781 TI - [Dehydroepiandrosterone (DHEA)--structure, clinical importance and the role in human body]. AB - DHEA and DHEA-S are hormones synthetized primarily by the adrenal cortex. The levels oh this hormones are systematically decreased, beginning from the fourth life decade. The levels of this hormones are also abberrated as a consequence of divorce systematical diseases like cardiovascular diseases, skeletal diseases, diabetes mellitus or obesity. This hormones, probably, have antiaheromatic facilities. There are also data suggesting their influence on stimulation of immunological system. It is already confirmed that the levels of this hormones are modified in congenital function disorders that are present in different diseases, like Alzheimer diseases, and oral administration of DHEA can improves the memory. Presumably DHEA-S have also anticarcinogenic facilities. The levels of this hormones can be also a marker monitoring the course of pregnancy. There are still a lot of discrepancies between results of different studies and it is very difficult to describe their role in human body. Because their levels are decreased with ageing process, this observation makes the researchers call them as the "youth hormones". PMID- 11875782 TI - [Oxidative stress in glomerulonephritis]. AB - The aim of this review is to focus on the possible role of reactive oxygen species (ROS) in the pathogenesis of glomerulonephritis (GN) and to review the potential mechanisms of this ROS-mediated renal injury. This paper examines the literature which demonstrates ROS as primary mediators in GN, responsible for modifications of glomerular permeability to proteins, development of morphologic lesions and alteration of glomerular haemodynamics (reduction in glomerular blood flow and glomerular filtration rate). In glomeruli, ROS are generated by both infiltrating cells (neutrophils, monocytes) and resident glomerular cells (mesangial and endothelial cells and podocytes). The participation of ROS in glomerular damage was proved in many experimental studies by detection of products of oxidant injury in renal tissue or urine and by demonstration of a protective effect of antioxidants in those lesions. PMID- 11875784 TI - 'It's like Enron'. FPA, other physician practice management companies hear death knell. PMID- 11875783 TI - [Melatonin as an antioxidant]. AB - This review describes the structure and properties of melatonin. The interaction of melatonin with reactive oxygen species and its protective action in relation to DNA, lipids and proteins are presented. The effect of melatonin on antioxidant and prooxidant enzymes is discussed, too. PMID- 11875785 TI - Uneasy truce. Catholic-run public hospital to walk fine line. PMID- 11875789 TI - Attorneys general take on HMOs. PMID- 11875790 TI - Getting on the right track. Better supply management means a better bottom line, and healthcare is catching on to software that maximizes efficiency. PMID- 11875791 TI - When more means less. Utah gets federal approval limiting Medicaid services to some to provide a basic health package to 25,000 low-income uninsured. AB - Utah hospitals are cautiously optimistic about a controversial new Medicaid project that would expand health insurance coverage to 25,000 of the state's low income workers but would not cover hospitalization. The project marks the first time the federal government has let a state offer a Medicaid benefit plan that fails to cover hospitalization and reduces benefits for some enrollees in order to expand access for others. PMID- 11875792 TI - States search for ways to trim back spending. Officials consider rollbacks, delays in attempt to minimize effects of federal Medicaid cuts. PMID- 11875793 TI - Rising to the challenge. PMID- 11875794 TI - Internally displaced persons. AB - There were estimated to be over 20 million internally displaced persons (IDPs) at the end of 1999, a number that surpasses global estimates of refugees. Displacement exposes IDPs to new hazards and accrued vulnerability. These dynamics result in greater risk for the development of illness and death. Often, access of IDPs to health care and humanitarian assistance is excluded deliberately by conflicting parties. Furthermore, the arrival of IDPs into another community or region strains local health systems, and the host population ends up sharing the sufferings of the internally displaced. Health outcomes are dismaying. From a health perspective, the best option is to avoid human displacement. WHO contributes to the prevention of displacement by working for sustainable development. Placing health high on the political agenda helps maintain stability, and thereby, reduce the likelihood for displacement. Primary responsibility for assisting IDPs, irrespective of the cause, rests with the national government. However, where the government is unwilling or unable to provide the necessary aid, the international humanitarian community must step in, with WHO playing a major role in the health sector. There is consensus among the partners of the World Health Organization (WHO) that, in emergencies, the WHO must: 1) take the lead in rapid health assessment, epidemiological and nutritional surveillance, epidemic preparedness, essential drugs management, control of communicable diseases, and physical and psychosocial rehabilitation; and 2) provide guidelines and advice on nutritional requirements and rehabilitation, immunisation, medical relief items, and reproductive health. If the vital health needs of IDPs--security, food, water, shelter, sanitation and household items--are not satisfied, the provision of health services alone cannot save lives. Community participation is essential, and community participation implies bolstering the assets and capacities of the beneficiaries. PMID- 11875796 TI - The concept of assisted management of large-scale disasters by horizontal organizations. AB - Management of large-scale disasters is impeded by inadequately designed organizational infrastructure. The vertical organizational structures of most agencies responding to disasters contribute to a poorly integrated response, especially when collaboration, information sharing, and coordination are required. Horizontal (or lateral) organizations have assisted traditionally vertical civilian and military agencies by enhancing their capacity to operate successfully in complex human emergencies and large-scale natural disasters. Because of the multiagency and highly technical multidisciplinary requirements for decision-making in chemical and biological disasters, similar horizontal management options must be considered. PMID- 11875795 TI - External chest compression in the management of acute severe asthma--a technique in search of evidence. AB - Compelling anecdotal evidence exists for the potentially lifesaving benefits of mechanical external chest compression (MECC), but no published trials of the technique exist. The history and technique for MECC are discussed and illustrated by a case report. Although the technique is not discussed in the Resuscitation Guideline 2000, and the need for it within the intensive care unit has reduced, the use of MECC will have its greatest impact when initiated in the prehospital setting for patients suffering from severe, sudden-onset, asphyxic asthma. PMID- 11875797 TI - Emergency health surveillance after severe flooding in Louisiana, 1995. AB - INTRODUCTION: In disaster situations, timely surveillance systems that provide illness, injury, and mortality information to public health officials and hospitals are essential for planning and evaluating interventions. OBJECTIVES: To describe flood surveillance methodology, the impact of the event on hospitals, and the number of daily patient visits due to selected illnesses and injuries before, during, and after severe flooding in southeastern Louisiana in May 1995. METHODS: Survey of disaster-area hospitals regarding flood impact. Emergency department surveillance of injuries and illnesses for the week before, the two days during, and the week after the flood. RESULTS: There occurred an increase in the number of persons who drowned or were injured that presented to the moderately affected hospitals during the storm, but there was no increase in visits for gastroenteritis to any group of hospitals. Services were disrupted in more than half of hospitals. The severely affected hospitals had the least variation in the average number of daily visits. None of the drownings were reported by those hospitals that reported severe service disruption. CONCLUSIONS: Data should be collected from all hospitals in or near disaster areas, even if they were not directly affected by the disaster. Public education about the danger of drowning during flash flooding must be improved. The Louisiana experience emphasizes the need for a disaster-preparedness plan for rapid surveillance of illnesses and injuries. PMID- 11875798 TI - Medical relief personnel in complex emergencies: perceptions of effectiveness in the former Yugoslavia. AB - Humanitarian medical assistance and intervention during the civil war in Bosnia and Croatia was felt by national health workers to be relatively ineffective (2.8 on a 5-point Likert scale), compared to other forms of humanitarian assistance such as medical supplies (4.4/5) and non-medical materials (3.9/5). Bosnian physicians treating civilians noted that the most helpful types of personnel were surgeons and emergency physicians. This study suggests that assessment of personnel needs at the recipient level, in addition to standard relief assessments, is required early in models of complex emergencies. This study supports existing epidemiological models of complex emergencies, especially when high trauma-related mortality and morbidity are likely to occur. PMID- 11875799 TI - Mass gathering medicine: a predictive model for patient presentation and transport rates. AB - INTRODUCTION: This paper reports on research into the influence of environmental factors (including crowd size, temperature, humidity, and venue type) on the number of patients and the patient problems presenting to first-aid services at large, public events in Australia. Regression models were developed to predict rates of patient presentation and of transportation-to-a-hospital for future mass gatherings. OBJECTIVE: To develop a data set and predictive model that can be applied across venues and types of mass gathering events that is not venue or event specific. Data collected will allow informed event planning for future mass gatherings for which health care services are required. METHODS: Mass gatherings were defined as public events attended by in excess of 25,000 people. Over a period of 12 months, 201 mass gatherings attended by a combined audience in excess of 12 million people were surveyed throughout Australia. The survey was undertaken by St. John Ambulance Australia personnel. The researchers collected data on the incidence and type of patients presenting for treatment and on the environmental factors that may influence these presentations. A standard reporting format and definition of event geography was employed to overcome the event-specific nature of many previous surveys. RESULTS: There are 11,956 patients in the sample. The patient presentation rate across all event types was 0.992/1,000 attendees, and the transportation-to-hospital rate was 0.027/1,000 persons in attendance. The rates of patient presentations declined slightly as crowd sizes increased. The weather (particularly the relative humidity) was related positively to an increase in the rates of presentations. Other factors that influenced the number and type of patients presenting were the mobility of the crowd, the availability of alcohol, the event being enclosed by a boundary, and the number of patient-care personnel on duty. Three regression models were developed to predict presentation rates at future events. CONCLUSIONS: Several features of the event environment influence patient presentation rates, and that the prediction of patient load at these events is complex and multifactorial. The use of regression modeling and close attention to existing historical data for an event can improve planning and the provision of health care services at mass gatherings. PMID- 11875800 TI - A comparison of rural and urban Emergency Medical System (EMS) personnel: a Texas study. AB - INTRODUCTION: In treating accident victims, actions by the Emergency Medical Personnel (EMP) at the scene may be the difference between life or death, full recovery or permanent disability. Development of selected profiles based on locale of services, tenure, and paramedic certification will provide valuable insight into the diversity within the Emergency Medical Services (EMS) profession. Not only will these profiles enable administrators to improve their recruitment, training, and retention of the emergency medical workforce, it potentially could enhance the quality of health care in the community. POPULATION: Emergency medical personnel attending a statewide conference in Texas in late 1996 (n = 425). HYPOTHESES: 1) There is no difference between the profiles of urban and rural EMP; 2) There is no difference between the profiles of urban EMP with < 9 years of experience and those with > or = 9 years of experience; 3) There is no differences between the profiles of rural EMP with < 9 years of experience and those with > or = 9 years of experience. 4) There is no difference between the profiles of urban EMP with paramedic certification and those without certification; and 5) There is no difference between the profiles of rural EMP with paramedic certification and those without certification. METHODS: EMP attending the conference completed 425 survey instruments measuring five demographic features, five work-related features, and two psychological features. Survey instruments were included in each registrant's conference package. Completed surveys were deposited anonymously in labeled receptacles throughout the statewide conference site. Data collection ceased at the end of the conference. Discriminant analysis identified distinct profiles for the urban and rural EMP. RESULTS: The urban EMP, more than rural subjects, was younger (mean = 36 years), more likely to be compensated 100% for their services, had a higher level of education (mean = 13.8 years), and reported a lower level of burnout. Urban EMP with < 9 years of experience tended to be younger, male, married, and reported less burnout. Urban paramedics were more likely to be compensated 100% for their services, and had achieved a higher level of education. The rural EMP with < 9 years of experience were less likely to be paramedic, reported lower burnout scores, and was younger. The rural EMP without paramedic certification was more likely to be a volunteer, and have had fewer years of service. CONCLUSIONS: In Texas, locale of service (urban or rural), length of tenure as an EMP (> or = 9 years), and paramedic certification appear to be significant factors that define the EMP population in Texas. PMID- 11875801 TI - The effects of Hurricane Mitch on a community in northern Honduras. AB - INTRODUCTION: Hurricane Mitch was an event described as one of the most damaging recent natural disasters in our hemisphere. This study examined its effects on a community of 5,000 residents in northern Honduras. METHODS: Survey responses of 110 attendants at an ambulatory clinic 4 months after the event were analyzed. Correlates were established between demographic and housing characteristics and morbidity and mortality. RESULTS: The availability of food, water, and medical care decreased significantly immediately after the hurricane, but by four months afterward returned to baseline values. Residents reported emotional distress correlated with the loss of a house or intrafamilial illness or mortality. Diarrheal illnesses more commonly were found in households with poor chronic access to medical care. The use of cement block housing correlated with availability of food or running water, with access to medical care and vaccinations, and with a reduced frequency of diarrhea or headaches in the immediate post-hurricane phase. CONCLUSIONS: Improvements in housing construction appear to be the most effective preventive measure for withstanding the effects of future hurricanes in tropical regions similar to northern Honduras. PMID- 11875802 TI - Topical mafenide hydrochloride aqueous spray in initial management of massive contaminated wounds with devitalized tissue. AB - Since at least WWII, some open, contaminated wounds involving massive soft tissue injury and vascular damage have resulted in "irreversible shock," despite prompt rescue, hemorrhage control, and blood and fluid replacement, without signs of clinical infection. In animal studies, survival time was related statistically to the dosage of Clostridium perfringens in multicontaminated explosive wounds. Survival time was lengthened by the application of some topical antibacterial agents, but actual recovery was achieved only with topical mafenide hydrochloride solution aqueous spray, which resulted in negative clostridium. perfringens cultures. Although not related statistically to survival time, the mafenide hydrochloride spray also controlled the Pseudomonas aeruginosa in these wounds. Mafenide hydrochloride had the American trade name of Sulfamylon from about 1942 until 1998, when another pharmaceutical company patented Sulfamylon as the trade name for mafenide acetate, a weaker antibacterial agent. However, mafenide hydrochloride still is available from chemical companies. Mafenide hydrochloride solution spray has been used successfully in treatment of patients with severe by contaminated wounds and deep burns, and its use in initial care should be revisited. PMID- 11875803 TI - Principles of disaster management. Lesson 13. Organizational development. AB - Change in any organization is difficult. Relief organizations constantly are evolving and changing form to adapt to different needs, demands, and environment. As the phases of a disaster evolve, adjustments must be made by relief organizations to meet the changing needs. The sequential processes used to manage change include recognition and diagnosis of the problem, identification of alternatives, recognition of limiting conditions, selection of a strategy for change, and implementing and monitoring the change. The techniques used to effect change may be classified as structural, management, or technological. Changes can occur in division of labor, content of the work, relationships with other workers, supervisory and/or technical skills, operations, and decision-making hierarchy. Approaches can be mandated from the top, worked out jointly by management and personnel, or implemented by the affected personnel. Implementation of changes has two dimensions: timing and scope. Whenever changes are implemented, the impact of the changes must be monitored and the effects compared with what was expected. PMID- 11875804 TI - [Clinical utility and safety of diagnostic thoracoscopy]. AB - The increasing use of thoracoscopy performed under local anesthesia has made contributions to the diagnosis of pleural disease with effusion. During the past 7 years, we have performed 100 such thoracoscopy procedures using a flexible fiberoptic bronchoscope. On the basis of our clinical findings, we are able to discuss the utility and safety of this procedure. The causes of pleural effusion were carcinomatous pleurisy in 72 cases, tuberculosis pleurisy in 15 cases, infection without tuberculosis in 4 cases, malignant pleural mesothelioma in 8 cases and one case of asbestosis. The success rate of thoracoscopic pleural biopsies were 97% for carcinomatous pleurisy, 100% for malignant pleural mesothelioma and 86% for tuberculosis pleurisy. This procedure was performed with no serious effect on blood pressure, oxygen saturation, monitored ECG or BGA data, and with no serious complications. Therefore, we concluded that this method is very useful for the diagnosis of pleural effusions and has few complications. PMID- 11875805 TI - [Cavernostomy and simultaneous removal of fungus ball of pulmonary aspergilloma]. AB - Between 1991 and 2000, 21 patients (16 male and 5 female) underwent 28 cavernostomies for the treatment of pulmonary aspergilloma. The median age was 59.4 years (range 37-85 years). The mean %VC was 59.6 (range 30.4-91.2), and the mean FEV 1.0 was 1.51 ml (range 0.64-2.67 ml). The mean body mass index was 17.6 (range 12.7-23.2). The most common complaint was hemoptysis. The underlying lung disease was tuberculosis in 17 cases, atypical mycobacteriosis in 2, and unclassified in 2. All cases had been diagnosed as complex aspergilloma. The mean surgical duration was 136 minutes (range 85-203 min.) and the mean blood loss during surgery was 242 ml(range 5-810 ml). No death or major complications occurred in the postoperative course. During follow-up, 4 patients died of massive hemoptysis, cancer, respiratory failure or an unknown cause. Relapses of aspergilloma occurred in 9 patients (42.9%). Recavernostomy was performed safely on 5 patients. In conclusion, although the relapse rate of aspergilloma was high after cavernostomy, safe reoperations were performed. Cavernostomy is thus an effective treatment in high-risk patients. PMID- 11875806 TI - [Clinical features in twenty-three patients with paragonimiasis westermani]. AB - We assessed the clinical features in twenty-three cases diagnosed as paragonimiasis westermani from April 1988 to May 2000 in our clinic at Miyazaki Medical College Hospital. Twenty-one patients had eaten wild boar or crab meat. Their symptoms included productive cough, chest pain and hemosputum. The findings of radiography and computed tomography (CT) were nodules, masses, infiltrative shadows and pleural effusion. The percentages of eosinophils in the peripheral blood were elevated in 16 of 23 patients, and the serum IgE values were elevated in 12 of 18 patients. Serologic tests, which are very helpful for diagnostic purposes were performed on the blood sera of all patients, and on the pleural effusions of five patients. It is often difficult to diagnose paragonimiasis westermani in patients in whom eosinophilia or increased serum IgE is not present. However, paragonimiasis westermani should also be considered as a possible diagnosis when abnormal shadows are found in chest radiographs and chest CT. PMID- 11875807 TI - [A case of sarcoidosis involving multiple organs that showed significant improvement as the serum KL-6 level fell after steroid therapy]. AB - Pulmonary sarcoidosis was diagnosed by transbronchial lung biopsy in a 27-year old man admitted to our hospital in September 1996. In October 1999, bilateral parenchymal findings on chest radiography and the results of pulmonary function tests had significantly deteriorated compared with those of three years before. Since skin, heart, liver, and spleen lesions were also detected, this patient was treated with corticosteroids. The lung parenchymal lesions and the pulmonary function as well as the abnormal findings in other organs showed significantly improvement along with a decrease of the KL-6 level after steroid therapy. PMID- 11875808 TI - [Pulmonary infection due to Mycobacterium szulgai, a group 2 nontuberculous mycobacterium]. AB - A 37-year-old male was admitted to our hospital because of abnormal findings seen in a chest radiograph from a physical examination. There was nothing unusual about his past history, and he had been healthy. Chest radiography showed an infiltrative shadow with a cavity in the right upper lobe. A smear test of the sputum was negative but a culture was positive for a mycobacterium, which was identified as M. szulgai. A diagnosis of atypical pulmonary mycobacteriosis caused by M. szulgai. The patient was treated with isoniazid, rifampicin and ethambutol, which gradually improved the infiltration shadow, and his sputum cultures became negative for mycobacteria. In recent years, the frequency of atypical mycobacteriosis has increased up to 10% to 15% of all mycobacterial infections. In Japan, M. avium complex and M. kansasii account for more than 90% of such infections, so other isolates are rare. Most previous cases of atypical mycobacteriosis were secondary infections associated with underlying disease. In this patient, however, we describe a rare case of pulmonary infection due to M. szulgai appearing in a healthy male without underlying disease. PMID- 11875809 TI - [Pulmonary hyalinizing granuloma with massive infiltration of lymphocytes]. AB - A 39-year-old man was admitted to our hospital for examination of multiple nodules and infiltrates on a chest radiograph. His chest HRCT revealed multiple nodules with or without thick- or thin-walled cavities. Specimens obtained by video-assisted thoracoscopic biopsy showed bundles of hyalinized collagen fibers, some of which contained accumulated plasma cells in the center. The nodules were surrounded by massive lymphoid cells which formed germinal centers. These findings are compatible with pulmonary hyalinizing granuloma. The lymphoid cells looked uniform in some areas and had infiltrated along the bronchioles and small vessels and into the intralobular septa in a manner resembling pulmonary lymphoma or pseudolymphoma. The findings suggested that pulmonary hyalinizing granuloma may overlap pulmonary lymphoma. The disease has shown no progression for four years although no treatment has been given. PMID- 11875810 TI - [A case report of pulmonary blastoma--biphasic pulmonary blastoma]. AB - We report a biphasic pulmonary blastoma in a 77-year-old man. In a routine chest radiographic examination, in the left upper lung field, a massive shadow that had been entirely absent one year before was detected. The preoperative imaging films showed a 10 x 8 cm, well-circumscribed solid tumor in the upper lobe of the left lung. A preoperative clinical diagnosis of primary lung cancer was considered. The intraoperative findings were that the tumor had invaded the visceral pleura with adhesion to the parietal pleura. Left upper lobectomy with lymph node sampling was performed, and since invasion of the pulmonary artery wall was confirmed, complete removal of tumor was not possible. The postoperative diagnosis was biphasic pulmonary blastoma. The patient was treated with radiotherapy up to a limit of 50 Gy, covering the area around of the left hilum. At the latest follow-up, 1 year postoperatively, the patient was clinically and radiologically free of the disease. PMID- 11875811 TI - [A case of multiple pulmonary fibroleiomyomatous hamartoma]. AB - A 50-year-old woman had been followed up at another hospital since 1996 because of multiple nodular shadows in both lung fields on chest radiography and CT. She was admitted to our hospital on 27 July 1999 to undergo further examination for the chest shadows, which had become enlarged since 1998. One of them was surgically removed by using video-assisted thoracoscopy. Its pathological features showed a mass composed of spindle-shaped smooth muscle cells, located adjacent to the bronchiolar wall and connected with the bronchiolar smooth muscle. Moreover, the mass had cystic spaces covered only with monolayered bronchiolar epithelium. Therefore, the mass was considered to be a hamartomatous proliferation of bronchiolar smooth muscle rather than a metastatic tumor of uterine leiomyoma. This case was thought to be important, since there are few reports concerning the pathological features of multiple pulmonary fibroleiomyomatous hamartomas. PMID- 11875812 TI - [Pulmonary granulomatous lesion with severe chest pain and hemoptysis: a case report]. AB - We encountered a rare case of pulmonary granulomatous lesion accompanied with severe chest pain and hemoptysis. A 42-year-old man visited our hospital complaining of hemosputum. A chest radiograph showed a nodular shadow in the left lower lung field. Further examinations including fiberoptic bronchoscopy, bronchoalveolar lavage and transbronchial lung biopsy did not suggest a diagnosis. During the course of his illness, he suffered an episode of severe chest pain which could be controlled only by intravenous morphine chloride (10 mg). The chest radiograph at the time showed a broad infiltration in the left lower lung field. However, the lung perfusion scintigram taken 2 days before demonstrated decreased blood flow in the same field. We waited for the infiltration in the chest radiograph to diminish and then performed partial resection of the left lower lobe, thus terminating both hemosputum and chest pain. Histological examination showed a cavitary lesion in the periphery of the lung, surrounded by large numbers of infiltrating plasma cells and lymphocytes, among which were many hemosiderinladen macrophages. A small amount of mycelium, considered to be Nocardia or fungus, was seen in the cavity wall. These findings may indicate that an infection had contributed to the formation of a hemorrhagic granulomatous lesion, and that this lesion caused chest pain mainly because of the pleuritis and the decrease in the local pulmonary circulation. PMID- 11875813 TI - [A case of tuberous sclerosis with pulmonary lymphangiomyomatosis]. AB - A 52-year-old woman complained of exertional dyspnea. She had two sons with tuberous sclerosis and presented with facial angiofibroma and bilateral renal angiomyolipomas. Her chest radiograph demonstrated diffuse reticular shadows and chest CT revealed numerous well-defined cysts outlined by thin walls. Pulmonary lymphangiomyomatosis was diagnosed by transbronchial biopsy that showed multifocal proliferation of immature smooth muscle cells located in the walls of some bronchi, and in alveolar walls and pulmonary vessels. Immunoreactivity for SMA and HMB 45 was present in the same cells, but immunostaining for estrogen and progesterone receptors was negative. Much is still unknown about pulmonary lymphangiomyomatosis and pulmonary involvement in tuberous sclerosis. Further studies, such as gene analysis, will clarify the relationship between these diseases. PMID- 11875814 TI - [A case of Legionella pneumonia successfully treated intravenously with both erythromycin and ciprofloxacin]. AB - A 54-year-old man was admitted to our hospital complaining of a high fever and watery diarrhea. The chest radiograph on admission revealed a homogeneous consolidation of the left upper lobe. Laboratory findings included proteinuria, oligouria, hematuria, myoglobinuria, hyponatremia, and serum CPK elevation. On the basis of these findings, a tentative diagnosis of Legionella pneumonia was made. He was treated with sulbactam/cefoperazon and erythromycin, but his high fever remained and the consolidation shadow deteriorated. He was therefore given both erythromycin and ciprofloxacin intravenously. After several days the fever had returned to normal, the appearance of the chest radiograph had improved, and his symptoms were quickly relieved. This case suggests that intravenous administration of ciprofloxacin and erythromycin can be an effective treatment against Legionella pneumonia. PMID- 11875815 TI - [A case of polymyositis complicated with pulmonary tuberculosis concerned with steroid resistance due to rifampin administration]. AB - A 63-year-old woman in whom polymyositis had been diagnosed since 1994 was treated with ten mg of prednisolone. In 1998, she contracted pulmonary tuberculosis simultaneously with acute exacerbation of the polymyositis. The polymyositis was then treated with 40 mg of prednisolone, and the pulmonary tuberculosis, with isoniazid (INH), rifampin (RFP) and ethambutol (EB). However, the polymyositis was worsening, possibly because the administration of RFP had induced steroid resistance. Because of this, a large amount of steroid was considered necessary for the treatment of the polymyositis (PSL 80 mg/day). An additional administration of cyclosporine (CyA) made possible a decrease in the amount of steroid given. RFP has a potency to induce the enzyme p450, which has the effect of metabolizing steroids or CyA, thus shortening the half-life of these agents. When a case of polymyositis is superimposed with pulmonary tuberculosis, it should be remembered that RFP may shorten the half-life of any steroid or CyA that is administered as treatment. PMID- 11875816 TI - [A case of combined squamous cell carcinoma and aspergilloma arising in a cyst wall]. AB - A 76-year-old man in whom interstitial pneumonia and diabetes mellitus had been diagnosed complained of bloody sputum in August, 1998. Chest radiography disclosed irregular shadows in the left lower lung field. Chest computed tomography (CT) scans revealed a cyst and a small nodular lesion in the left S6 segment. Although primary lung cancer was suspected, we did not detect any malignant cells in the transbronchial lung biopsy specimen. CT scans in January 2000 showed a ball-like shadow in the thick-walled cyst in the left S6 segment. Cytologic examination of the sputum and the bronchial lavage fluid from the left B6 revealed squamous cell carcinoma. Left lower lobectomy and mediastinal lymph node dissection were performed. Pathological examination revealed that moderately differentiated squamous cell carcinoma had extensively invaded the wall of the cyst in the left S6 and S10 segments, and was accompanied with aspergilloma. Abnormal thickening of a cyst wall may in some cases suggest the presence of lung cancer. PMID- 11875817 TI - [A case of Pneumocystis carinii pneumonia with pulmonary cysts and increased level of serum KL-6]. AB - In August, 1999, a 46-year-old man with fever, cough, and dyspnea was admitted to a hospital. On the basis of the clinical and radiographic findings, bacterial pneumonia was suspected. Antibiotics were not effective, because of atypical lymphocytes in the peripheral blood and positive anti-human T-cell leukemia virus antibody, and he was transferred to our hospital. A chest radiograph and a CT scan revealed bilateral ground-glass opacities with huge multiple cysts. Intensive treatment of Pneumocystis carinii pneumonia associated with human T cell leukemia was unsuccessful. Pneumocystis carinii was found in the bronchoalveolar lavage fluid. Human T-cell leukemia and Pneumocystis carinii pneumonia were diagnosed. In this case, numerous pulmonary cysts were progressing rapidly, the largest cyst being 8.7 cm in diameter, and the largest cyst in our experience either in clinical practice or in reading of the literature in Pneumocystis carinii pneumonia. The maximum serum KL-6 was markedly increased to 15,200 U/ml, which is the highest level reported for Pneumocystis carinii pneumonia. PMID- 11875818 TI - [Natural food additives given new function by enzymatic reaction]. PMID- 11875819 TI - [Formation of ethyl carbamate in umeshu (plum liqueur)]. AB - Samples of umeshu, a Japanese plum liqueur made from unripe plums, shochu and crystal sugar, were stored under fluorescent light, in the dark and in the refrigerator. The amount of ethyl carbamate formed in umeshu exposed to light or room temperature was larger than that in the dark or at low temperature. The amount of ethyl carbamate formed in umeshu to which cyanide had been added was larger than that in the absence of added cyanide. Thus, the amount of ethyl carbamate formed in the umeshu was increased by not only light and higher temperature, but also cyanide. Samples of model alcoholic beverages were stored under various conditions using red, yellow and blue cellophanes. The amount of ethyl carbamate formed in the model alcoholic beverage with blue cellophane was larger than in the cases of red and yellow cellophanes. It was found that the amount of ethyl carbamate formed in the model alcoholic beverage was increased by light in the wavelength range of 375-475 nm. PMID- 11875821 TI - [Comparison of carotenoid components between GM and non-GM papaya]. AB - We compared the carotenoid profile in GM papaya (Sunup) line to that of a non-GM one (Sunset). First, to identify major carotenoids in papaya, large-scale extraction was carried out with methanol. HPLC analyses of the methanol extracts revealed that both papayas mainly contained 5 pigments and no apparent difference was observed in the HPLC profiles. On the basis of LC/MS data and photodiode array spectra, beta-carotene (3), lycopene (2), beta-cryptoxanthin (1), and beta cryptoxanthin myristoyl and lauroyl esters (4 and 5) were identified as major carotenoids. It is well known that most carotenoids are labile, so a rapid analysis with precautions to avoid decomposition was developed to quantify their contents in the original fruits. Frozen samples of the fruits were sliced and a piece (about 2 g) of fruit was cut out and lyophilized. After extraction of the piece with methanol containing an anti-oxidant, BHT, the extract was further partitioned with hexane and methanol. Finally the contents of the main carotenoids in the hexane fraction were analyzed by HPLC. The contents of total carotenoids (sum of 1-5) and beta-cryptoxanthin (1, 4 and 5) in GM papaya fruit were estimated to be 0.764 +/- 0.056 and 0.604 +/- 0.051 (mumol/g), respectively and those in non-GM fruit were 0.883 +/- 0.145 and 0.705 +/- 0.098 (mumol/g), respectively. These differences are not statistically significant. PMID- 11875820 TI - [Determination of 2,4,6-tri-tert-butylphenol and related compounds in foods]. AB - An analytical method has been developed for the determination of 2,4,6-tri-tert butylphenol (TTBP) in foods. TTBP was determined by GC/MS (SIM) after extraction from food samples using a steam distillation technique. The developed method was able to determine simultaneously 2,4-di-tert-butylphenol (2,4-DTBP), 2,6-di-tert butylphenol (2,6-DTBP), 3,5-di-tert-butylphenol (3,5-DTBP) and 2,4-di-tert pentylphenol (2,4-DTPP). The method was applied to analyze the residues of the 5 phenolic compounds in 101 food samples purchased from markets. TTBP was found in some samples of meat, liver and fish (muscle) at the levels of trace (tr)-0.50 ng/g, tr and tr-1.83 ng/g, respectively. 2,4-DTBP was found in some samples of vegetables, meat, liver, fish (muscle) and fish (viscera) at the levels of 1.4 10.6 ng/g, 2.7-26.4 ng/g, tr-34.2 ng/g, tr-21.6 ng/g and tr, respectively. 2,6 DTBP was found in some samples of fish (muscle) and fish (viscera) at the levels of tr-3.9 ng/g and tr, respectively. 3,5-DTBP and 2,4-DTPP were not found in any of the analyzed samples. PMID- 11875822 TI - [Determination of streptomycin and dihydrostreptomycin in meat by liquid chromatography/mass spectrometry]. AB - A sensitive and selective method using liquid chromatography-electrospray mass spectrometry (LC-ESI-MS) for the determination of aminoglycoside antibiotics, streptomycin and dihydrostreptomycin, in meat has been developed. The LC separation was performed on a TSK-gel Super ODS column (10 cm x 2 mm i.d.) using 5 mmol/L heptafluoro-n-butyric acid (HFBA)-acetonitrile (88:12) as the mobile phase at a flow rate of 0.18 mL/min. The positive ionization produced typical [M + H]+ molecular ions of both drugs (streptomycin m/z 582; dihydrostreptomycin m/z 584). The calibration graphs for streptomycin and dihydrostreptomycin were rectilinear from 0.25 to 25 ng with selected ion monitoring (SIM). The drugs were extracted with 1% metaphosphoric acid, and the extracts were added to 2 mL of 0.1 mol/L heptanesulfonic acid. The solution was cleaned up on a Bond Elut C18 (500 mg) cartridge. The recoveries of streptomycin and dihydrostreptomycin from swine and bovine muscle fortified at 0.2 microgram/g were 73.2-82.6%, and the detection limits were 0.01 microgram/g for both drugs. PMID- 11875823 TI - [Preparation of HPLC test solutions for organic impurities in aluminum lakes of food red no. 40 (allura red AC) and food yellow no. 5 (sunset yellow FCF)]. AB - The HPLC determination of organic impurities in Food Red No. 40 aluminum lakes (R 40Als) as directed by Japan's Specifications and Standards for Food Additives, 7th Ed. (JSFA-VII), has problems, such as reproducibility and low recovery. ICP analyses suggested that the problem was caused by the aluminum in the test solution. In the improved method for preparation of the test solution, aluminum was precipitated as a hydroxide gel by boiling with 1% aqueous ammonia. After centrifugation, the supernatant was used for the HPLC analysis of the organic impurities in the lakes. Recoveries of organic impurities were more than 85% from R-40Al spiked at the 0.1 and 1.0% levels of R-40. The proposed method was also adapted for Food Yellow No. 5 aluminum lakes. PMID- 11875824 TI - [Multiresidue analysis of organophosphorus pesticides in vegetables and fruits using dual-column GC-FPD, -NPD]. AB - We investigated simultaneous and consecutive analytical methods for pesticide residues in large numbers of vegetable and fruit samples. Extraction of the sample with acetonitrile was followed by a salting-out step using a graduated cylinder. Co-extractives were removed by gel permeation chromatography (GPC) and the pesticide eluate was separated to 2 fractions. Firstly, the second fraction (85-125 mL) was passed through a silica-gel mini-column. After putting a Florisil mini-column before the silica-gel mini-column, the first fraction (55-85 mL) was passed through the tandem mini-columns, which were then eluted with 15 mL of 50% acetone/petroleum ether. The eluate was subjected to dual-column GC with dual FPD (P mode, column Rtx-OPPesticides) and NPD (column Rtx-200) detectors. Recoveries of 63 organophosphorus pesticides from fortified spinach, tomato, apple and strawberry, ranged from 71 to 126% with RSD values of 1-18%, except for the RSDs of omethoate, isoxathion, and pyraclofos (20% <). Detection limits of pesticides were 0.5-2 ng/g. Surveillance of pesticides in vegetables and fruits was carried out by using the present method. From 20 out of 30 samples, 15 pesticides (39 in total) were detected. The results indicated that the present method can be applied as an efficient and reliable tool for monitoring organophosphorus pesticide residues in vegetables and fruits. PMID- 11875825 TI - Occurrence of clostridia in commercially available curry roux. AB - The occurrence of clostridia was investigated in a total of 60 commercially available curry roux samples. Clostridia were isolated from 37 (62%) samples, and Clostridium perfringens was isolated from 7 (12%) samples. The isolates of C. perfringens did not produce enterotoxin. The frequency of occurrence was higher by the enrichment broth culture detection method than by the agar plate or pouch method. These findings suggest that enrichment broth culture is necessary for the detection of clostridia. PMID- 11875826 TI - [Clean-up procedure with ion-exchange mini column in the analysis of flusulfamide in agricultural products by HPLC]. AB - A clean-up procedure with an ion-exchange column in the analysis of flusulfamide by HPLC was examined. Pesticide in the sample was extracted with methanol following liquid-liquid partition with ethyl acetate. The ethyl acetate fraction was cleaned up with silica gel column chromatography. The eluate from the silica gel column was further cleaned up with SAX + PSA mini column, then determined by HPLC. Carotenoids and interfering peaks were removed by washing the combined mini columns with 10 mL of 20% acetone-containing n-hexane and 5 mL of acetone, and flusulfamide was eluted with 35 mL of acetone. PMID- 11875827 TI - Estimation of preservative concentrations in foods and their daily intake based on official inspection results in Japan in fiscal year 1998. AB - The mean concentration and daily intake of five preservatives were estimated based on the results of an analysis of 89,927 samples of food obtained in official inspections by Japanese local governments in fiscal year 1998. The mean concentration of benzoic acid was 9.5% of the allowable limit, and those of dehydroacetic acid, p-hydroxybenzoic acid, propionic acid, and sorbic acid were 1.5%, 5.7%, 1.7%, and 23.9%, respectively. Daily intake levels of these preservatives per person estimated from the concentration and daily consumption of foods were 6.23 mg, 0.0303 mg, 1.02 mg, 8.10 mg, and 25.0 mg, respectively, and assuming a body weight of 50 kg, the amounts of benzoic acid, p hydroxybenzoic acid, and sorbic acid consumed were 2.5%, 0.2%, and 2.0% of their acceptable daily intakes, respectively. These values were similar to those obtained based on the results of the official inspections in fiscal years 1994 and 1996. PMID- 11875828 TI - [An introduction to predictive microbiology]. PMID- 11875829 TI - [Interpretation of food poisoning by natural poisons and chemical substances (1980-1997)]. PMID- 11875830 TI - [The approach to pesticide performance in analytical procedure]. PMID- 11875831 TI - [New food additives applying for the designation and the revision of standards for use]. PMID- 11875832 TI - Medical staffing. Exclusion zones. AB - The UK has a lower ratio of doctors to population than most other developed countries. The government's pledge to increase the number of doctors by 9,500 by 2004 will represent only a marginal increase. Reliance on doctors from other parts of Europe, as envisaged in the NHS plan, is not realistic. European countries have provided few doctors for the UK. The position of non-European overseas doctors in the NHS, and the low numbers achieving consultant status, needs a radical review. Institutionalised racism must be addressed. Future medical workforce planning must provide exchange and development opportunities for developing countries. PMID- 11875833 TI - Open space. Take five. PMID- 11875834 TI - Nurse training. Teenage hang-ups. AB - A survey of acute trusts' nurse cadet schemes in England showed a wide variety in entry requirements and qualifications offered. Most schemes offered successful students a place on a nurse training scheme. Legal issues surrounding 16-year olds working in hospitals need to be clarified. Standardisation of entry requirements and course qualifications is required. At present, a student could complete a cadet course in one part of the country and not be eligible for nurse training elsewhere. PMID- 11875835 TI - [Undifferentiated connective tissue disease: clinical and serological profile of 578 patients followed for five years: disease course, prognosis and therapy]. AB - INTRODUCTION: Evolution of immunopathological diseases is usually slow and progressive. The term the undifferentiated connective tissue disease (UCTD) is used to describe the phase preceding a defined connective tissue diseases (CTD). AIMS: The objective of this work was evaluate the clinical and serological profile of patients with UCTD, who had been followed between 1994-1999. They have investigated the frequency and the type the developed autoimmune diseases from UCTD. PATIENTS: A total of 578 UCTD patients were evaluated. RESULTS: In 143/578 patients (24.7%) with the UCTD differentiated to systemic connective tissue diseases (28 systemic lupus erythematosus, 26 mixed connective tissue disease, 19 progressive systemic sclerosis, 3 polymyositis/dermatomyositis, 45 Sjogren syndrome, and 22 systemic vasculitis). 86.7 percent (124/143) of the systemic connective disease developed in first two years of UCTD. The condition of 435/578 (75.2%) remained UCTD after 5 years, among them in 82 patients with UCTD was regression of the symptoms. The presence of the fever and anti-DNS antibodies correlated with SLE (P = 0.0104, Fisher exact test), arthritis/arthralgia and anti-RNP antibodies with MCTD (P = 0.0302), Raynaud phenomenon and ANA positivity with PSS (P = 0.0144), xerostomia/xerophtalmia and anti-SSA/SSB antibodies with Sjogren syndromes (P = 0.0144). CONCLUSIONS: The UCTD in our patients seem to represents an dynamic phase, one part of the patients show progression to definite connective tissue diseases, one part show regression, and on part of the patients stay in UCTD phase. PMID- 11875836 TI - [Endoscopic carpal tunnel release: long-term results]. AB - INTRODUCTION: Based on the literature data in the last ten years the endoscopic carpal tunnel release seems to be an advantageous alternative method in the treatment of carpal tunnel syndrome. AIMS: Based on minimum 5 year follow-up the authors report the long-term results of endoscopic surgery introduced since 1993. PATIENTS AND METHODS: On 46 hand of 42 patients endoscopic procedure was performed. Thirty-two patients returned to follow-up, they filled out questionnaire, underwent physical examination and 15 patients had electrophysiological test. RESULTS: Twenty-eight patients were without any complaints and symptoms, 3 patients had thenar atrophy. One patient was reoperated on because of residual symptoms. The authors make a comparison between their data and the great experienced Chow's results and they review the literature data of open procedure. CONCLUSIONS: They conclude that the long-term results of the endoscopic surgery are similar to the outcome of the open procedures. The electrophysiologic tests have not always any relationship with the patient's complaints and symptoms. PMID- 11875837 TI - [Benign pulmonary and tracheal tumors in our biopsies]. AB - INTRODUCTION: Benign lung and trachea tumors account for only a small percentage of a bronchologist's and a histopathologist's everyday practice. AIMS: The authors describe the clinicopathological features some of these tumors because they may be confused with malignant conditions. To avoid misdiagnosis of malignancy the knowledge of bronchologic and histologic appearance of these tumors is important. RESULTS: In the past three years 1453 cases were examined and 14 cases of benign tumors were diagnosed, which is 1% of the total number of biopsies and 2.68% of the tumor cases. The diagnoses were: granular cell tumor (Abrokosoff's tumor), neurilemmoma, hemangioma, endobronchial hamartoma, and papillary adenoma. Two of the reported tumors: granular cell tumor and papillary adenoma appeared in the trachea. Tracheal granular cell tumor and bronchial hemangioma were incidental findings beside bronchial carcinoma. CONCLUSION: To our best knowledge this is the fifth reported case of the coexistence of Abrikossoff's tumor with bronchial carcinoma. PMID- 11875838 TI - [Double-blind peer review]. AB - The peer review process in medical sciences is much debated. The method is not yet evidence based. Who are the reviewers and how do they perform? They are "independent" experts. As such they are bound to be involved in research similar to that outlined in the manuscript. Very often they are contestant in the same race. That is why the author consider the double blind method, where the author is blinded and the reviewer masked from each other's identity, the best choice. Nevertheless in very many scientific journals of high quality and envied impact factors do not blind the identity of the authors. Science is a race for fame, self accomplishment and also a means to get grants for pursuing the scientific research. Only success is able to provide resources for expensive scientific research. There is no fail-safe method against bias in grant giving and editorial process. In the tidal wave of electronic information it is mandatory to help in differentiating between signal and noise in science. Peer review is the best method to protect readers from the trash of uncontrolled publications in medical science. PMID- 11875839 TI - [A language reformer and physician: Jozsef Polya (1802-1873)]. PMID- 11875840 TI - [The medical laboratory: the topic segment is expanded in Praxis]. PMID- 11875841 TI - [D-dimer test in the general practice laboratory]. PMID- 11875842 TI - [Basedow disease. From subtotal to total thyroidectomy]. AB - BACKGROUND: Surgical treatment of Graves' disease is based on modern pathophysiolic understanding and adequate surgical tactic and technique. STUDY: This is an audit from one institution about 81 consecutive, prospectively documented patients, undergoing subtotal (remnant < 6 g) or total thyroidectomy, by the technique of capsular dissection. RESULTS: Patients were female in 89%, aged 11-79 (median 35) years. They constitute 9% of all thyroid operations, i.e. 47% of those performed for hyperthyroidism. Indications were persistence and recurrence of disease, orbitopathy, large goiter, nodule formation, refusal of radioiodine, preconception control. Preoperative treatment was thyrostatic (97%), iodine (87%), propranolol (50%). During the study period use of total thyroidectomy increased from 0% to 87% (p < 0.0001), and the remnant size decreased from median 3.5 g to 0 g (p < 0.0001). Goiter weight was 20-255 g, median 70.28 (23%) patients had concomitant colloid or adenomatous nodules, 5 (6%) had an incidental microcarcinoma, and 3 (4%) had a clinical papillary or follicular carcinoma, 1 patient had a parathyroid adenoma. Mortality was 0; surgical morbidity was early postoperative haemorrhage (n = 2 (2.5%)), permanent nerve palsy (1 patient with recurrence after previous resection (1.2%; 0.6% of nerves at risk)); no case of permanent hypoparathyroidism occurred. Functional results: In 6 patients (8%; 15% of those with subtotal resection) recurrent hyperthyroidism developed, 1 month to 8 years postoperatively, necessitating reablative treatment (surgical in 2 instances). 1 further patient developed preclinical hyperthyroidism 11 years postoperatively. Postoperative thyroxine substitution was found to be inadequate in 16/73 (20%) patients, as demonstrated by suppressed or elevated TSH values. Recurrence-free survival at 10 years was 100% in patients with a remnant measuring < 3 g, vs. 55.8% in those with a remnant size > or = 3-6 g (p = 0.002). Ophthalmopathy improved in 33/47 (70%) and deteriorated in 5 (10%) patients postoperatively, independently of the procedure (total or subtotal). CONCLUSIONS: Extensive or total thyroidectomy is necessary for late recurrence-free survival.--Variation of the spontaneous remnant function may occur and contribute to inadequate substitution.--(Near) total thyroidectomy has a low morbidity and is considered the treatment of choice, also with the theoretical advantage of complete autoantigene removal.--Surgical progress is based on capsular dissection with fine preparatory operative technique. PMID- 11875843 TI - [Radiofrequency ablation of supraventricular re-entry tachycardia: a "new life" with a successful and complications-free method]. AB - The aim of this retrospective study was to evaluate the success rate and the satisfaction of patients after radiofrequency ablation of supraventricular tachycardias. 94 consecutive patients, 62 with AV-node-reentry-tachycardia (AVNRT) and 32 with AV-reentry-tachycardia (AVRT) were included. Success rate of ablation was 98% for AVNRT and 94% for AVRT. Recurrence rates were 11% and 9% respectively. The interval between the first tachycardia and ablation was very long (mean 17 +/- 14 years) and prior drug therapy was successful in only 13% of patients. Minor complications were observed in 12% and only one major complication (AV-fistula) was treated surgically. No pacemaker had to be implanted due to a complete AV-block. After ablation 96% of patients were "strongly satisfied" or "satisfied" with the procedure and their relief of symptoms. In addition limitations in all investigated activities (everyday-life, work, sport, hiking, travelling, sexual activity) were significantly reduced (all p < 0.005). PMID- 11875844 TI - [Percutaneous transpedicular vertebroplasty for treatment of osteoporosis-induced vertebral fractures]. AB - Osteoporosis plays an increasing medical, social, and economic role in our society. Vertebral body fractures are the most common fractures, along with fractures around the hip joint, and fractures of the distal radius due to osteoporosis. An osteoporotic vertebral compression fracture can lead to incapacitating back pain and immobility and often requires in-hospital treatment. Vertebroplasty is an effective procedure consisting of the percutaneous injection of polymethylmethacrylate (PMMA) cement into vertebral bodies. Preliminary studies showed promising clinical results with this new technique. As there is a high incidence of cement leakage during the injection of the cement, we prefer to perform the procedure under computed tomography control with a fluoro-CT-option. Furthermore, we recommend that the procedure is only done in collaboration with a spine surgeon. In case of a neurological complication, immediate decompression of the spinal canal is mandatory. Vertebroplasty is a very promising, effective and safe method for the treatment for osteoporotic vertebral compression fractures. Based on our preliminary results, this method can be recommended for a more widespread clinical use. PMID- 11875846 TI - [Risk factors for mortality and locomotor ability 6 months after proximal femoral fracture]. PMID- 11875845 TI - [Exanthema with emphasis of body flexure points 10 hours after administration of amoxicillin, "baboon syndrome"]. PMID- 11875847 TI - ["Who is interested in fast and wished-for death--thoughts of a health economist"]. PMID- 11875848 TI - Epidemiological concepts regarding disease monitoring and surveillance. AB - Definitions of epidemiological concepts regarding disease monitoring and surveillance can be found in textbooks on veterinary epidemiology. This paper gives a review of how the concepts: monitoring, surveillance, and disease control strategies are defined. Monitoring and surveillance systems (MO&SS) involve measurements of disease occurrence, and the design of the monitoring determines which types of disease occurrence measures can be applied. However, the knowledge of the performance of diagnostic tests (sensitivity and specificity) is essential to estimate the true occurrence of the disease. The terms, disease control programme (DCP) or disease eradication programme (DEP), are defined, and the steps of DCP/DEP are described to illustrate that they are a process rather than a static MO&SS. PMID- 11875849 TI - Economic aspects of disease monitoring with special reference to bovine paratuberculosis. AB - Monte Carlo simulation models were used to evaluate the feasibility and potential results of a proposed national survey of the prevalence of bovine paratuberculosis (PTB) in dairy herds in Norway. The expected herd prevalence was assumed to be 0.2% in the simulations. Infected herds were classified as detected if 1 animal was sero-positive. With a sample size of 6000 herds at least 1 truly infected herd was detected in 99% of the iterations. The low sensitivity of the ELISA test, the assumed low herd prevalence, the typical low within-herd prevalence of PTB and the small herd sizes in Norway all present problems in detection of the disease. The results showed that the ratio between false positive herds and true positive herds detected had a median of 70:1. At the assumed herd prevalence of 0.2% and a cost/test of 70 NOK the median cost of detecting 1 infected herd was approximately 900,000 NOK. If 2 positive reactors were needed to classify a herd "infected" the median cost of detecting 1 infected herd was 5,055,000 NOK. Our results suggest that a randomized national prevalence survey would not be feasible, due to the low probability of detecting infected herds and because of the high number of false-positive reactions that would be expected. PMID- 11875850 TI - Surveillance of ruminant diseases in the Nordic countries. AB - All the Nordic countries have a basis for their surveillance and disease control in ruminants in national legislation and regulations listing notifiable diseases of concern to the countries. The Nordic countries are a disease-free zone comparing to other parts of the world and the aim of the surveillance is to keep that status and be able to document it. Following is a short summary from each country. PMID- 11875851 TI - Disease surveillance strategies in swine. PMID- 11875852 TI - Surveillance of fish diseases in the Nordic countries. AB - Due to the increasing importance of disease problems in the fish farming industry and the impact disease may have on both feral and farmed fish in the Nordic countries, monitoring and surveillance on diseases have for many years been considered to be of socioeconomic importance. All the Nordic countries have a national legislation as basis for their surveillance and disease control in aquatic animals and regulations listing notifiable diseases of concern to the countries. The list of diseases vary between the countries. In addition, Denmark, Finland and Sweden are ruled by Directive 91/67/EEC as regards placing on the market of aquaculture animals and products. The surveillance for viral diseases in all the Nordic countries has mainly been based on the testing procedures given in the EU Commission Decision 96/240/EC. PMID- 11875853 TI - Disease recording systems and herd health schemes for production diseases. AB - Disease recording of cattle is compulsory in Sweden and Norway. Sweden and Denmark also have mandatory disease recording for swine, whereas Finland and Norway only have compulsory recording of infectious diseases. Both compulsory and voluntary systems are data based, the first ones developed in the 1970's. Disease recording at pig slaughtering is somewhat older. The veterinary practitioner, and often also the farmer, can report treated cases as well as fertility disturbances to the systems. Disease recording at slaughter is carried out by veterinarians and inspection officers. The data bases are handled by the veterinary authorities or the agricultural organisations in each country. Costs are defrayed by the authorities and/or the agricultural industry. The farmers receive periodic reports. Data are stored for three to ten years, often longer. Affiliation to animal health schemes for cattle or swine is voluntary. In Sweden and Denmark (cattle) they are run within the scope of government regulations. Affiliation to animal health programmes may also be demanded by organisations within the agricultural industry. These organisations are also responsible for the administration of the programmes. Costs to take part in herd health schemes are covered by the farmers them-selves. In certain cases, grants are received from agricultural organisations, authorities, or the European Union. Recording of diseases and the format of animal health schemes in the Nordic countries are described here in order to illustrate the possibilities to compare data between countries. PMID- 11875855 TI - Why do we record disease? PMID- 11875854 TI - A concept for a decision support system based on practical experiences from a national disease emergency. The Dutch experience. AB - In 1997-1998, the Netherlands experienced a large epidemic of classical swine fever (CSF). The magnitude of this epidemic stressed the role information systems could play in supporting the management during an eradication campaign. The enforcement of an eradication strategy can become very complicated, especially with large epidemics, due to time pressure and many different procedures that have to be executed at the same time. The application of comprehensive information systems may result in more control over the process and in a relief of the operational management. After a brief description of the Dutch epidemic the authors provide an overview and the general application of four different types of information systems, classified as decision support systems. The application of these information systems in animal disease control is illustrated by providing concepts for a system architecture for transaction processing, management and executive information support and decision support. The application of a data warehouse as part of this systems architecture is explained. The eradication of CSF from the Netherlands was complicated by several factors. It is important to notice that information systems cannot prevent these factors. However, information systems can support disease control authorities in controlling these factors. PMID- 11875857 TI - Geographical Information System (GIS) as a tool in surveillance and monitoring of animal diseases. AB - In the veterinary epidemiology, the advantage of mapping the locations of farms and other facilities with animals is obvious. In an outbreak of a disease it could make the management of the situation easier, and it could also provide a tool to evaluate different strategies to prevent the spread of infectious diseases. This paper aims to describe and give an overview of the possibilities and potential uses of a Geographical Information System (GIS) in the field of surveillance and monitoring of animal diseases. The following areas in which GIS and special GIS-functions could be incorporated are presented: recording and reporting information, epidemic emergency, cluster analysis, modelling disease spread, and planning control strategies. Different sources of data; geographical data, farm locations and disease information, used in the development of the GIS at the National Veterinary Institute in Norway are thoroughly described in the paper. Further, it presents a few examples where the GIS has been applied to studies of epidemiology and surveillance of animal diseases in Norway, which shows the significant value of GIS in these areas. At the same time, the incorporation of GIS in this field shows the scarcity of the data available, which should encourage improvement in the data recording and the quality of the registries. PMID- 11875856 TI - Relationships between animal health monitoring and the risk assessment process. AB - Risk assessment is part of the risk analysis process as it is used in veterinary medicine to estimate risks related to international trade and food safety. Data from monitoring and surveillance systems (MO&SS) are used throughout the risk assessment process for hazard identification, release assessment, exposure assessment and consequence assessment. As the quality of risk assessments depends to a large extent on the availability and quality of input data, there is a close relationship between MO&SS and risk assessment. In order to improve the quality of risk assessments, MO&SS should be designed according to minimum quality standards. Second, recent scientific developments on state-of-the-art design and analysis of surveys need to be translated into field applications and legislation. Finally, knowledge about the risk assessment process among MO&SS planners and managers should be promoted in order to assure high-quality data. PMID- 11875858 TI - Documenting freedom from disease and re-establishing a free status after a breakdown. Rabies. AB - Rabies reappeared in Finland in the spring of 1988 after a 29-year absence. This time rabies occurred in sylvatic form and the major species involved was the raccoon dog. During the outbreak 1988-89 66 animals were diagnosed rabid. Vaccination of cats, cattle and horses was strongly recommended and vaccination of dogs was compulsory in the outbreak area. A field trial was started on oral immunisation of raccoon dogs and foxes against rabies using baits containing rabies vaccine strain. The outbreak area and a wide buffer zone were baited three times. Finland was declared free of rabies again in 1991. Oral vaccination campaign with vaccine baits has been organised along the south-eastern border once a year since the beginning of 90s. Continuous surveillance and epidemiological screening is necessary to detect any new outbreaks of rabies at an early stage. PMID- 11875859 TI - [Clarifying informed consent]. PMID- 11875860 TI - [Feminine acne: dermatologic disease or endocrine disease?]. AB - Acne is a problem of the pilo-sebaceous follicle caused by the conjunction of three factors: seborrhea, follicle obstruction, and follicle inflammation. The key element, seborrhea, is under androgenic control. Acne in women is also influenced by developments and modifications in genital life, as well as by hormonal contraceptive and replacement therapies. Acne is rare prior to puberty, when it may indicate endocrine disease. At puberty, acne is quasi-physiological, because of the relative hyperandrogenism induced by the andrenarche preceding pubarche, as well as by the relative shortage of estrogens and progesterone during the first menstrual cycles. Other signs of hyperandrogenism, such as menstrual cycle difficulties and excess weight, which favor a hormonal origin, must be sought in cases of persistent or late-onset acne in adults. There is a mirror image of puberty during the peri-menopausal period, but with decreased seborrhea, so acne is rare. Finally, a tumoral origin must be sought in the rare cases of acne occurring after menopause. Hormonal investigation of acne should not be systematic, but is justified during prepuberty when other symptoms are associated with acne that resists well-conducted dermatological treatment. The therapeutic approach should be primarily dermatological, but hormone-oriented treatment should be considered when such therapy fails, or in the presence of other signs of hyperandrogenism. Sometimes the association of isotretinoin and an anti-androgen treatment are necessary to effectively treat such acne. Finally, particular attention must be paid to contraceptive therapies and hormone treatments, which can induce or aggravate acne, especially during the peri menopausal period. PMID- 11875861 TI - [I.U.D. degradation in utero as a function of time: the phenomena of calcification. Radiocrystallographic and scanning electron microscopy study]. AB - OBJECTIVE: A study was undertaken in order to determine the frequency of the process of calcareous deposition, to clarify the nature of the deposits, their arrangement along the IUD and their possible influence on the appearance of metrorrhagias. MATERIAL AND METHODS: A total of 408 coopper IUD's belonging to four standard models were observed by means of optical microscopy at low magnifications. The most typical samples were studied under the scanning electron microscope and deposits analyzed by crystallography. The data were analysed statistically. RESULTS: It was possible to determine both the frequency and the importance of the process of calcification according to the model of IUD and to the bearing time, to demonstrate a preferential location of concretions and to suspect a sensibility to the phenomenon variable according to the model. It was also possible to clarify the stages of the process of calcification, to describe its most characteristic aspects and to identify the essential component of calcareous deposits. CONCLUSIONS: Calcification concerns roughly over 50% of IUD's in utero. It occurs preferentially on the fundic portion of an IUD. Obviously, the minimal time required for the phenomenon to start as well as to increase its intensity are submitted to strong individual variations. Concretions are composed of vaterite, a distinct variety of carbonate of calcium. They do not seem to be the primary cause of possible metrorrhagias. PMID- 11875862 TI - [The ancient Alexandria school of medicine]. AB - A famous medical school was established in old Alexandria during the third century BC. Although mainly Greek in essence, and following the Hippocratic teachings, it was heavily tainted by the medical practices of ancient Egypt. Anatomy was particularly advanced due to the possibility of dissecting the human body. The most important Alexandrian physicians were Herophilus and Erasistratus. Many graduates of this medical school traveled and practiced throughout the Mediterranean basin. Galen, the famous Roman physician studied in Alexandria before practicing in Rome. His teachings and writings survived well into the sixteenth century and formed the basis of more modern medical practices during the renaissance. These writings were conserved partly by Christian monks and partly by Arab and Jewish scholars of the middle ages. The medical school of Alexandria was still active until late in the 3rd century AD. However, it slipped slowly into oblivion after the fire of 389 AD, which also devastated its famous library. PMID- 11875864 TI - [Gestational trophoblastic diseases. Apropos of 105 cases]. AB - The authors report a retrospective study of 105 observations of gestational trophoblastic diseases managed at the university clinic of obstetric gynecology I (Pr Chaoui). Of this study, one listed 72 cases of complete mole hydatiforme with 5 cases of sacrofetal pregnancy. The invasive mole is found in 4 cases and the choriocarcinoma in 24 cases. The general frequency of this pathology is of 1/770 pregnancies. The age of our patients varies from 15 to 52 years with an average age 27 years and the multiparity is found in 50% of the cases. 103 patients (95.5%) consulted for metrorrhagia associated pelvic pains in 31 cases (30%). The toxic syndrome was present in 20 patients (12%) with a preeclampsy in 6 cases (5.71%). The physical examination showed a very increased uterus of size in 92 cases (87.5%) associated adnexal mass in 37 cases (35.2%). The diagnosis was especially echographic in the totality of the cases associated or not with a proportioning of plasmatic beta HCG or prolans urinary. The treatment of the trophoblastic disease varies simple endo-uterine aspiration (85%) until the chemotherapy treatment (32.4%), the hysterectomy was indicated in a third of the cases. The evolution of the non complicated mole hydatiforme was good in 100% of the cases, it quasi totality of the invasive moles presented a complete remission. Among the 24 choriocarcinoma, we deplore 4 deaths in a table of pulmonary, hepatic and cerebral metastases. In order to improve the forecast of these diseases, the diagnosis must be early with an adequate treatment and a rigorous monitoring. PMID- 11875863 TI - [Comparison of predictive values of inhibins A and B, and plasma estradiol in IVF patients treated with GnRH agonists and recombinant FSH]. AB - The objectives of this study were to determine the mean plasma inhibin A and B kinetics in normoovulatory patients treated by GnRH agonist and rec-FSH and to compare their predictive value with that of plasma estradiol on retrieved oocytes number and pregnancy rate. The study was carried out retrospectively in 36 normoovulatory IVF patients stimulated by GnRH agonist from D21 and rec-FSH from D2 of the following cycle. Two groups of 18 patients (having obtained or not a pregnancy) were paired for age and cause of infertility. Estradiol was measured by direct immunoassay. Inhibin A and B were measured by ELISA (Serotec Limited, Oxford, UK). Inhibin A was correlated with estradiol until P-2 (P0: day of follicles aspiration). Inhibin B rose earlier, its concentration reached its maximum at P-5. Plasma estradiol and inhibin A kinetics were not different among pregnant and nonpregnant women. There was a trend for a broader plasma Inhibin B surface under the curve in pregnant than in nonpregnant women. Estradiol and Inhibin A were correlated to the oocytes number at the end of stimulation (P-2 and D10). Inhibin B was correlated earlier than estradiol and inhibin A, at P-8 and D7. Plasma inhibin A did not demonstrate a better predictive value than estradiol neither of the oocytes number nor the chance of pregnancy. Inhibin B could be useful for early decision of cycle cancellation or stimulation adjustment. PMID- 11875865 TI - [Benign mesothelioma of the uterus. Clinical and histopathological aspects and evidence of its mesothelial origin. Apropos of 1 case]. AB - Benign mesothelioma is a rare tumor nearly always find in relation to the genital tract. We report the case of a 47-year, old woman admit in the gynecologic department for metrorragia. The tumor was composed of irregular tubules between smooth muscle cells, lined by flattened or cuboidal cells without nuclear atypia or mitoses. The tumor cells express cytokeratins and vimentin. Histologically, differential diagnosis can be made with adenocarcinoma or vascular proliferation like lymphangioma, hemangioma or angiomyoma. Clinical outcome is always favourable without recurrence or malignant transformation. PMID- 11875866 TI - [Psychological aspects of the diagnosis of chromosomal aberrations in the first trimester]. AB - Over the past ten years, rapid and quite considerable developments have been made in antenatal screening and foetal scanning. This progress had led to the early identification of increasingly minute malformations and a more accurate diagnosis of a number of foetal pathologies. For some years now, parents have been offered the possibility of an early screening of the Down's syndrome during the 1st quarter scan. If the risk proves to be a major one for the child, the mother can have an amniocentesis to determine if a chromosomic anomaly does in actual fact exist. At the same time, as medical science advances, it is important to take into consideration the psychological impact these screening tests can have on parents and in particular mothers. The purpose of this article is to highlight the real-life experience of parents and examine the reality of parental anguish within the specific framework of 1st quarter chromosomic screening. We will also look at the role psychologists, psychiatrists and psychoanalysts are called on to play with patients and medical teams. PMID- 11875867 TI - [Peripartum dilated cardiomyopathies: is there a correlation with sexually transmitted diseases?]. AB - A revue of literature about peripartum cardiomyopathy; a disease of unknown pathogenesis. Some retrospective studies suggest a relation with sexually transmitted diseases. Other risk factors were observed. Diuretics and digoxin are used in the treatment. Cardiac transplantation is the final solution but the affection could appear again. A database must be started with epidemiologic information to understand this disorder and its correlation with sexually transmitted diseases. PMID- 11875868 TI - [Fetal pulse oximetry in 2001]. PMID- 11875870 TI - [Contribution of exogenous LH during ovarian stimulation: for a targeted indication]. PMID- 11875869 TI - [GnRH antagonists]. PMID- 11875871 TI - [Contribution of exogenous LH during ovarian stimulation: against a systematic indication]. PMID- 11875872 TI - [Adjuvant treatment for breast cancer. Results of the consensus conference of Saint-Gall (February 2001)]. PMID- 11875873 TI - [Is it necessary to make of the laws of voluntary interruption of pregnancy in France flexible again? Gynecol Obstet Fertil 2001: 29: 137-8]. PMID- 11875874 TI - [Management of early symptoms of acute septic hip in neonates and infants- conservative or surgical?]. AB - The paper presents 41 cases of acute hip sepsis in 36 patients, age ranging from 5 to 66 days. 32 cases were treated at the Institute of Polish Mother Hospital and 4 at the Pediatric Surgery Department of the Medical School of Lodz. In 40% of the cases multifocal infections were noted. Septic arthritis was diagnosed basing on clinical symptoms, lab tests, ultrasound screening, X-rays and immunoglobulin scyntigraphy. Patients were divided into three groups according to treatment: group 1 was administered antibiotics and immobilization was applied (16 patients--19 hips), group 2 had a biopsy, was administered antibiotics and immobilization (5 patients--6 hips), group 3 had an arthrotomy performed, wound drainage, casting was applied and antibiotics were administered (15 patients--16 hips). Immobilization was maintained by means of a cast or Koszla's harness. Patients were qualified for surgery basing on ultrasound investigation. After completion of various modes of treatment patients were followed at the outpatient clinic of the orthopedic department. A complete assessment was performed 3-6 years after the first symptoms appeared. A full clinical examination (ROM assessment), X-ray investigations were performed. Hip dysfunction was assessed basing on X-ray images according to the Ho Choi classification. Conservative treatment seems to be sufficient in early stages of septic arthritis, with only minor morphological pathologies visible on ultrasound examination. Early arthrotomy seems to be the method of choice in septic arthritis treatment. Aspiration of the hip may not be possible due to solid consistency of pus. PMID- 11875875 TI - [The use of Bicontact hip prosthesis in treatment of coxarthrosis]. AB - The issue of a stabile, long lasting bone-implant interface is still very valid. One of the new models of uncemented hip prosthesis is the Bicontact prosthesis made by Aesculap. This four-part endoprosthesis offers two types of cups: threaded Munchen type and spherical Plasmacup type. The prosthesis stem is a wedged type stem, porous in the proximal type. The aim of this paper was to evaluate the value of Bicontact prosthesis in treatment of coxarthrosis. Our material consisted of 69 patients (38 female and 31 male) with 72 operated hips. Average age of the patients was 50.8 years. The mean follow-up was 24.5 months. Clinical and radiological evaluation according to Merle d'Aubine-Postel classification with Charnley's modification yielded 88% very good and good results. In 11 hips a similar complication was noted--fracture of the femur during final hammering of the prosthesis stem into the medullary canal. This was immediately repaired by circular wire fixation without any influence on the final result. This complication was observed during the first procedures. The learning curve of surgical technique allowed us to avoid this complication during subsequent procedures. We concluded that type of hip prosthesis was is very useful in in treatment of coxarthrosis. PMID- 11875876 TI - [Can samples from the cups of Weller endoprosthesis be the basis for analysis of the degree of biomaterial crystallinity under the influence of external factors?]. AB - The paper is a thesis verification. The thesis stated that samples from cups of Weller endoprosthesis can be the basis of biomaterial crystallinity analysis under the influence of external factors. Cup segments were deformed and exposed to X-rays. Basing on thermoanalytic analysis the authors prove that the cups (completely transformed and sterilized) cannot be used as research material, to be tested by application of external factors, to assess polyurethane morphology. An alternative reliable method has been proposed: direct investigation of the material used for cup production. These conclusions were reached as result of research done with a differential scanning calorimeter (heat-flux type) DCS 30 in Mettler TA-4000 thermoanalytic system. PMID- 11875877 TI - [Intramedullary Gamma nail fixation in the treatment of peritrochanteric fractures]. AB - The paper presents the authors' own experience in the treatment of periotrochanteric fractures. A clinical survey from 1998 to 2001 was carried out on 26 patients aged 21-72. The number of women was 20 (77%) and of men 6 (23%). This paper is an preliminary report. PMID- 11875878 TI - [Assessment of quadriceps muscle isometric strength following operative treatment of femoral shaft fractures]. AB - The authors investigated quadriceps muscle strength following operative treatment of femoral shaft fractures in 135 patients. Patients were categorized according to the kind of procedure performed: open Kuntscher intramedullary nailing--30 cases; Zespol--30 cases; AO plate osteosynthesis--47 cases and closed intramedullary nailing 28 cases. Two groups of patients were formed according to surgical technique: open method--107 cases and closed method--28 cases. 39 patients treated with an open method required re-operation. The best results were obtained in cases treated with closed technique--79.6%. The worst results were noted in the group requiring re-operation: 56.7%. The worst results were found in cases with comminuted fractures, and those located in the distal third of the femoral shaft. PMID- 11875879 TI - [Evaluation of clinical and radiological results of operative treatment of patellar fractures]. AB - The paper presents the clinical and radiological evaluation of surgical treatment of patellar fractures in the years 1984-1999. Th study group included 45 patients (18 female and 27 males). The observation period ranged from 2 to 6 years; on average 6 years. The most common procedure was tension band fixation (20 patients), followed by hemipatellectomy (14 patients) and patellectomy (11 patients). Radiological assessment revealed the highest number of excellent and very good results in patients treated with tension band fixation. No excellent results and 50% food results were noted in patients treated by partial patellectomy. The worst radiological resulted were noted in patients treated by total patellectomy. Clinical results were similar to radiological results: 75% of patients treated by tension band fixation had no pain, while the group treated by total patellectomy revealed only 27.3% excellent and very good results. PMID- 11875880 TI - [Surgical resection of primary soft-tissue sarcoma of the extremities]. AB - We evaluated 5 patients who had soft tissue sarcoma of the extremity treated by limb sparing surgery. All tumors were adjacent to bone, with no features of infiltration. According to the Surgical Staging System, all tumors were IIB. The mean follow-up was 13 months. To achieve wide resections of the tumors in such cases we recommend resections of the tumor with part of the bone, which is in direct contact with the tumor. Wide resection margins were achieved in all patients and none had local recurrence during the observation period. PMID- 11875881 TI - [Kramer's distal I metatarsal osteotomy in the treatment of halluces valgi]. AB - The paper presents the surgical technique for Kramer's distal I metatarsal osteotomy and the results of halluces valgi treatment in 23 patients. The mean age of the patients was 48.6 years and mean follow-up time was 4 years and 5 months. Final clinical and radiological evaluation was performed according to Kramer's scale. Excellent and good results were found in 54% of the cases, fair in 32% and bad in 14%. This simple extraarticular osteotomy, without use of any external immobilization yielding three-dimensional correction is a useful method for treatment of hallux valgus. PMID- 11875882 TI - [The USS (Universal Spine System) in the treatment of thoraco-lumbar spine fractures]. AB - The paper present the results of surgical treatment of 15 patients (11 males and 4 females--average age 35.3 years) with thoraco-lumbar spine fractures. Pre-op neurological disorders were found in 4 patients with narrowing of the spinal canal, including 1 case of paraplegia. The decrease of vertebrae height was on average 45%. All patients underwent surgical treatment with transpedicular fixation by means of USS (Universal Spine System). In 7 patients 4 vertebrae were instrumented, the rest of the patients had only 3 vertebrae instrumented. In this latter case arthrodesis was not performed. Post-op, the neurological symptoms receded completely in 3 patients and partially in the case with paraplegia. In 76% of the cases vertebra height was restored completely. Patients were allowed to stand on the 3-4 day post-op. Follow-up was 22 months, and the neurological status of the patients was stabile. No decrease in vertebra height was noticed. PMID- 11875883 TI - [HIV infection in the operating room conditions]. AB - Several cases of occupational HIV transmission during surgical procedures have been reported all over the world in the last few years. Orthopaedic surgeons are at particularly high risk for blood contamination and skin injuries. This article addresses the risk of HIV transmission in the surgical setting with a focus on specific risk factors: the prevalence of the infection in general population, the stage of the disease, virulence, inoculum size, the nature and frequency of blood contacts. It also shows how the last two factors depend on surgical specialty, duration and kind of the procedure, role of each person, number of needles used and amount of blood loss. The recommended safer surgical techniques as well as personal protective equipment are presented as the strategy to prevent health care worker--to patient transmission. The importance of compliance with universal precautions is pointed as a way to achieve a successful outcome in preventing infection with HIV. PMID- 11875884 TI - [Intervate dislocation of the distal interphalangeal joint of the annular finger in a child]. AB - The paper presents a good cosmetic and functional result of surgical treatment of distal interphalangeal joint dislocation of the annular finger in a 14 year old boy. The mechanism and pathology of this rare lesion of the hand are discussed. Factors affecting final outcome are: preserved joint cartilage, careful surgical approach and proper post-op treatment. PMID- 11875885 TI - [Snapping hip of internal origin]. AB - The paper presents an unusual case of internal snapping hip in an 18 year old female, due to abnormal iliopsoas tendon tension. Operative treatment with precision of the fibrous slip over the rim of the pelvis gave a very good result, with slight limitation of active hip flexion. Various opinions on the pathogenesis and treatment of this rare condition have been presented. Lack of snapping in a supine position during clinical assessment has been emphasized. PMID- 11875886 TI - [Late external iliac vein injury after total hip arthroplasty--a case report]. AB - The use of bone cement during total hip prosthesis implantation despite it's advantages triggers of some local and general complications. The paper presents and classifies most cement related complications, with special attention to vessel complications. This paper includes an external vein injury, which occurred 5 years after a total hip replacement procedure. Chronic erosion and mechanical stress of the iliac vein by a cement hook was the cause of this complication. It has been stress that most vascular complications are a result of imperfect surgical technique. PMID- 11875887 TI - [Sympathetically maintained pain syndrome--a case report]. AB - Sympathetically maintained pain syndrome (SMPS) is considered to be a clinical form of reflex sympathic dystrophy. It develops usually after trauma, and consists of continuous, burning pain with sympathetic component. We present a case of this syndrome affecting both hands, that developed in a patient 3 months after severe burn injury. Intravenous fentolamine test was used to recognise sympathetic transmission of the pain. The management included regional intravenous fentolamine blocks and orally administrated phenoxybenzamine. The former gave only temporary relief; after latter improvement lasted 2 months, but patient eventually failed to recover. Diagnostic and therapeutic considerations concerning this syndrome were underlined. PMID- 11875888 TI - [Classification of results of treatment of congenital clubfeet]. AB - The paper presents the comparison of late results of surgical treatment of congenital clubfeet using a number of assessment classifications. 61 feet in 46 patients treated between 1979-1990 with a mean follow-up time of 15.7 years. Strong correlations were noted between the various classifications, but none was found "ideal" or particularly useful in everyday clinical practice. PMID- 11875889 TI - [Struggle for freedom of the research dissemination]. PMID- 11875890 TI - [Free choice of hospital]. PMID- 11875892 TI - [Sports diving]. PMID- 11875891 TI - [Linguistic understanding of Nordic physicians]. PMID- 11875893 TI - [Pathological gambling]. PMID- 11875894 TI - [The letter of condolence--obsolete or beneficial?]. PMID- 11875895 TI - [Treatment of patients with postpolio syndrome in warm climate]. AB - BACKGROUND: Treatment in a warm climate of various patient groups, including patients with postpolio syndrome, is controversial. MATERIAL AND METHODS: 88 patients with postpolio syndrome (61 women) were recruited, stratified according to sex, age (<> 60 years old) and use/non-use of electrical wheelchair, and randomized to three groups. Group 1 (n = 30) underwent treatment in a rehabilitation centre in Tenerife for four weeks in November/December 1999. Group 2 (n = 29) was treated in two similar centres in Norway for the same period of time, while Group 3 (n = 29), the control group, followed their ordinary health care programme. All patients were tested at the start of the study and three and six months later with physical tests and several questionnaire and qualitative interviews. Patients in groups 1 and 2 were also tested after the rehabilitation period. RESULTS: Groups 1 and 2 improved significantly both in physical tests and subjective ratings. The positive effects in group 1 tended to exceed the positive effects in group 2, and the effects lasted longer. Six minutes walking distance in the two groups was 347 metres and 316 metres, respectively before the treatment period, 429 metres and 362 metres immediately after, and 431 metres and 356 metres three months later. Subjective rating of pain (VAS scale) was 42 and 43 respectively before treatment, 17 and 31 immediately after, and 28 and 44 three months later. In the control group, only minor changes were found. INTERPRETATION: The study seems to document a positive effect of treatment of patients with postpolio syndrome in a warm climate. PMID- 11875897 TI - [Evaluation of the new doctoral degree program at the Faculty of Medicine in Oslo 1993-98]. AB - BACKGROUND: In 1993, the doctoral degree programme in the Faculty of Medicine of the University of Oslo was substantially revised to include coursework and supervision of thesis work. PhD students were expected to complete their work towards the doctorate in three years, and funding was only provided for this period. MATERIAL AND METHODS: In spring 1999, all doctoral candidates, their supervisors and members of the adjudicating committees were invited to reply to a questionnaire with the purpose of evaluating the results of the new programme over the 1993-99 period. RESULTS: Only a few doctoral students had been able to obtain their degrees in three years, the defined length of the programme. The mean age for new PhDs was 38; however, physicians obtained their PhD at a later age than the other life scientists enrolled in the programme, and the percentage of PhDs with a medical background declined from 71% in 1993-95 to 51% in 1996-98. INTERPRETATION: The doctoral programme should be extended from three to four years. More physicians should go into research soon after graduating from medical school and more openings for postdocs should be created. More time for research in the university clinics is also needed. PMID- 11875896 TI - [Lyme neuroborreliosis in More and Romsdal]. AB - BACKGROUND: The broad variations in the clinical manifestation in Lyme borreliosis underline the importance of laboratory investigations in serum and cerebrospinal fluid. MATERIAL AND METHODS: We have studied patients with neurological signs compatible with Lyme neuroborreliosis, pleocytosis in cerebrospinal fluid and positive Borrelia serology in serum/cerebrospinal fluid analysed by ELISA. We have evaluated clinical characteristics, laboratory parameters, treatment effects, and incidence variations. RESULTS: We included 25 patients in the study. Isolated facial palsy was often seen, but clinical manifestations showed huge variation. Fourteen of 25 (56%) patients had positive Borrelia burgdorferi-IgM and IgG titres in cerebrospinal fluid despite negative tests in serum. The mean annual incidence rate in the county judged by notified cases to the Norwegian Surveillance System for Communicable Diseases (MSIS) was 4.4/100,000 in the period 1989-99 as compared to the national rate of 4.3/100,000 in the same period. In 1998, however, the annual incidence rate in the county was 8.8/100,000 as compared to the national rate of 4.1/100,000. INTERPRETATION: The diversity of symptoms and signs suggests a liberal attitude towards serological testing including CSF analyses. More and Romsdal is a high incidence region for Lyme borreliosis in Norway. The annual variation in incidence might reflect a changing prevalence of the tick vector along the Norwegian coastline. PMID- 11875898 TI - [Nose in the light of medical history]. AB - Vanity, it seems, is one of the qualities that separate us from other species; indeed, we may said to possess it in excess. Decorating body and apparel with all kinds of objects in order to look attractive goes a long way back in time; even Stone Age man knew how to make himself look smart. This very human trait has not abated over the millennia, but the perception of what looks good has changed a great deal, just like fashion, and no part of the human body has probably been of such significance in this respect as the nose. This article presents a brief review of nasal fashion. PMID- 11875899 TI - [Where are all the Viking helmets?]. AB - Based on archaeological finds and old Norse literature, this article describes the Scandinavian helmet tradition from the Bronze Age to the Viking Age, as well as the Viking culture, with special emphasis on weaponry, burial customs, and head protection. Contrary to what is commonly believed, metal helmets must have been used very infrequently by the Vikings. Only one Viking helmet has been retrieved in Scandinavia. Possible reasons for the wide-spread misunderstanding that the Vikings wore helmets are discussed. The archaeological profession must partly bear the responsibility for not correcting this misunderstanding. PMID- 11875900 TI - [Manuscripts don't burn]. AB - Mikhail Bulgakov (1891-1940) studied at Kiev University and qualified in medicine in 1916. He worked as a doctor for a few years, then chose a literary career. All his life he was sceptical to the Soviet system and used his satire against the regime. Because of that he met difficulties as an author and could not publish anything after 1927. He worked on his main work, The Master and Margarita, from 1928 until his death. The novel was not published in his lifetime. In 1966-67 an abridged and censored edition appeared. Not until 1973 was a complete edition published in Moscow, and it soon became a cult book in Russia. In the major part of The Master and Margarita, the scene is in Moscow between the two world wars, with Bulgakov using his own experience. Through subtle irony and wild humour he exposes the imperfections of this society. To obtain this, he employs the Devil with his suite of strange figures. The book is also a love story between the Master and Margarita. The Master is writing a novel about Pontius Pilate, and this narrative runs parallel to the Moscow story, so that the book puts an eternal perspective on human frailty. PMID- 11875902 TI - [Physical activity and health--recommendations]. AB - BACKGROUND: The health consequences of physical inactivity and the health gains of regular physical activity are briefly summarized. We present physical activity recommendations based on a review of the literature for the general adult population. METHODS: This report employs recognised international standards for establishing causal relationships between physical activity and disease, distinguishing between four different levels: Convincing, probable, possible and insufficient evidence. Convincing or probable evidence have provided the basis for the recommendations given. RESULTS: The total amount of physical activity (a combination of intensity, duration and frequency) is related to a number of health variables in a dose-response relationship. The preventive effect (the health gain) increases with increasing activity level, but the relationship is not linear. The greatest health gains are attained by those who are least physically fit. This applies even in old age. The health gain seems primarily to be dependent on the total energy expenditure, and less on the intensity. The dose response relationship between physical activity levels and health gains is a continuum that does not seem to have a lower limit. Thus, any activity can be said to be better than none. INTERPRETATION: We conclude that a minimum "target dose" that will yield substantial health gains for those who have been physically inactive for a long time, is activity of moderate intensity which corresponds to an energy expenditure of approximately 150 kcal (630 kJ) per day (or slightly more than 1,000 kcal (4.2 MJ) per week). This is equivalent to a daily walk of about 30 minutes. The activity can be divided into shorter intervals of physical activity during the course of the day, for instance intervals lasting 5-10 minutes. An increase in activity beyond this level will yield further health gains. The effect of exercise is just as good for older people as for younger people, and the same recommendations apply as to other adults. PMID- 11875901 TI - [Lou Andreas-Salome (1861-1937)--psychoanalytical and feministic contribution to understanding her biography]. AB - Lou (Louise) Andreas-Salome's life and work has preoccupied many biographers. The interest may have be sparked by her liaisons with many of the greatest men of her time. She had an intimate relationship with Friedrich Nietzsche in a period of great change for him. She was Rainer Marie Rilke's mistress for several years. And she pursued a close friendship and working relationship with Sigmund Freud in the latter part of her life. But her significance goes beyond these associations. She was a celebrated novelist and essayist in her own right, with ten novels and more than 50 essays, also on psychoanalytical subjects. She has been viewed as femme fatale, opportunist, feminist, radical, liberal, but also as a significant contributor to psychoanalytical thought. There have been two biographical approaches: a psychoanalytical approach focusing on her loss of father-figures and later difficult relationships with famous men, and a feministic approach accusing psychoanalysts of not contributing to insight, but belittling Salome's legitimate position. A fuller understanding may be obtained by integrating these two views. PMID- 11875903 TI - [Do Scandinavian physicians understand each other's languages?]. AB - BACKGROUND: The national medical journals in Denmark, Norway, and Sweden publish selected papers from the other journals in their original languages. We wanted to know how well Scandinavian general practitioners understand their neighbour languages and English, today the "lingua franca" in medical publishing. MATERIAL AND METHODS: 111 doctors (36 Norwegians, 32 Danes, 43 Swedes) completed a questionnaire in which they rated their own subjective understanding of Danish, Norwegian, Swedish, and English on a five point Likert scale. RESULTS: The Norwegian doctors rated their understanding of Danish significantly higher than Swedish (p < 0.01) and English (p < 0.01). The Swedish doctors rated English significantly higher than Danish (p < 0.01) and Norwegian (p = 0.01) and Norwegian significantly higher than Danish (p = 0.02). The Danish doctors did not rate the foreign languages differently. INTERPRETATION: Particularly because of the Swedish doctors' incomplete understanding of Danish and to some extent Norwegian, there is a risk that future communication between doctors in these countries will be conducted in the English language. PMID- 11875904 TI - [Tuberculosis among immigrants in Scandinavian medical journals]. AB - BACKGROUND: The decline in incidence of tuberculosis in Scandinavia has stagnated in the last decades because of a growing number of immigrants from high incidence countries. The aim of this study was to find out what has been published on tuberculosis among immigrants in Denmark, Norway and Sweden in the main medical journals in these countries from 1995 to 2000. MATERIAL AND METHODS: 23 papers on the topic were identified through PubMed searches. Eight papers were selected for a more detailed description of their content. RESULTS: The main medical journals in Denmark, Norway and Sweden have published papers on epidemiological aspects and clinical traits as well as the challenge for the health services of controlling tuberculosis in immigrants. The majority of patients are under the age of 40 and of African origin. They seldom infect the population of the host country. Extrapulmonary tuberculosis is relatively common. Control on arrival could be improved in all three countries. Considerable doctor's and patient's delay is not uncommon. INTERPRETATION: Medical journals are an important source for knowledge on tuberculosis among immigrants from high incidence countries. PMID- 11875905 TI - [Etiology of non-melanoma skin cancer]. AB - BACKGROUND: Most countries are experiencing an increased incidence of non melanoma skin cancer (basal cell carcinoma and squamous cell carcinoma). MATERIAL AND METHODS: Based on relevant literature found by search in the PubMed database, an overview of the present knowledge of the etiology and pathogenesis of non melanoma skin cancer is given. RESULTS: DNA damage caused by ultraviolet (UV) radiation from the sun is the main cause of non-melanoma skin cancer. Failure to repair these genetic alterations leads to unrestrained growth and ultimately tumour formation. In addition, ultraviolet radiation induces a relative immunosuppression in the skin that prevents cutaneous tumour rejection. UV specific mutations in the p53 tumour suppressor gene probably play an important role through both tumour initiation and cell selection. INTERPRETATION: Physicians should advise their patients to avoid unnecessary sun exposure and to use sun protection, including sun screen creams. PMID- 11875906 TI - [Drugs and advice to travellers]. PMID- 11875907 TI - [Paranoid personality disorders and political power]. PMID- 11875908 TI - [Some things were better before--how can we make use of them?]. PMID- 11875909 TI - [Norwegian medical students abroad]. PMID- 11875910 TI - [Norwegian medical students abroad]. PMID- 11875911 TI - [Revised national budget and peritoneal dialysis--all that glitters is not gold]. PMID- 11875912 TI - [The national health authority and the responsibility for cost-effective drug utilization]. PMID- 11875913 TI - [No one is avoiding the General Practice Research Committee]. PMID- 11875914 TI - [Doping]. PMID- 11875915 TI - [The family practice system--some important details]. PMID- 11875916 TI - [Medical history taking--only for the physician?]. PMID- 11875917 TI - [Cosmetic surgery on the edge]. PMID- 11875918 TI - [Mouth diseases--who does what?]. PMID- 11875919 TI - [Vitamin and fatty acid supplementation to infants]. PMID- 11875920 TI - [Use of statins in hospitals after myocardial infarction]. AB - BACKGROUND: The objective of this study was to register the frequency of statin prescription during the initial hospitalization for acute myocardial infarction and therapeutic intensity at follow-up. MATERIAL AND METHODS: A retrospective study among patients aged below 70 of both sexes with acute myocardial infarction, who survived the initial hospitalization at the University Hospital of Tromso during 1995-1998. RESULTS: 473 patients with acute myocardial infarction, 76% men, mean age 57.4 (range 33-70 yrs) were included. Statin treatment was started in 55% of the patients within discharge from the hospital. Total cholesterol (odds ratio 0.51, 0.41-0.64; 95% CI) and decreasing age (1.60, 1.21-2.10) were significant predictors for statin prescription. Statin treatment started during hospitalization increased gradually from 42% in 1995 to 91% in 1998 (p < 0.001) among patients with total cholesterol above 5.0 mmol/l at admission. 54% of the statin users achieved the treatment goal at their first outpatient control 8-12 weeks after discharge. INTERPRETATION: The proportion of statin users has increased satisfactory during 1995-98, but the therapeutic intensity is unsatisfactory compared to guidelines for cholesterol-lowering treatment of patients with acute myocardial infarction. PMID- 11875921 TI - [Surgery of femoral neck fractures--higher rate of osteosynthesis failure with the use of 4,5 mm screws compared to 6,5 mm screws]. AB - BACKGROUND: Displaced femoral neck fractures are usually treated with screw osteosynthesis or primary hip replacement (unipolar or bipolar). MATERIAL AND METHOD: In this retrospective study we compared the rate of early failure during two consecutive years using two different screw implants each year. The patient groups were demographically similar, and the relationships between the use of hip replacement and screws as primary treatment were similar in the two groups. In the first group (group A), two screws with a shaft and outer thread diameter both of 6.5 mm were used. In the second group (group B), two screws with shaft diameter of 4.5 mm and outer thread diameter of 7.3 mm were used. The number of patients operated with screws were 191 the first year and 177 the following year. RESULTS: The number of patients reoperated with a hip replacement within the first 100 days was 16 (8%) in group A and 29 (16%) in group B (risk difference 0.08; 95% CI 0.013-0.147). INTERPRETATION: We recommend that when two screws are used for fixation of femoral neck fractures, they should have a shaft diameter of at least 6.5 mm. PMID- 11875922 TI - [Liver resection--indications and results]. AB - BACKGROUND: Liver resection is an established treatment for malignancies like colorectal metastases and hepatocellular carcinoma. MATERIAL AND METHODS: Indications and outcomes of liver resection at the National Hospital, Oslo, Norway was studied retrospectively in 226 patients operated between 1977 and 1999. RESULTS: The main indication for surgery was colorectal metastases (n = 137). The frequency of liver resection for colorectal malignancies was < 1 per 100,000 patients per year in the hospital's catchment area. Other indications included hepatocellular carcinoma (n = 30), benign tumours like hemangioma (n = 14), and various primary and secondary malignant tumours. Reoperation due to postoperative complications was performed in 13 patients (6%). Total perioperative mortality defined as death before hospital discharge or within 30 days after discharge, was 3% (7/226). No perioperative deaths occurred among the 159 patients operated after 1987. Five year survival for patients operated for colorectal metastases and hepatocellular carcinoma were 29% and 24%, respectively. INTERPRETATION: The main indication for liver resection is colorectal metastases. Liver resection is a safe operation with potential curation for selected patients. PMID- 11875923 TI - [Hernia surgery in Norway]. AB - BACKGROUND: Inguinal hernia repair has undergone major changes during the last decade. This study aimed to explore the impact on treatment algorithms used in Norway. METHODS: A questionnaire was sent to all public hospitals in January 1999. RESULTS: 57 of 58 hospitals where inguinal hernia repairs were undertaken responded. Most repairs are undertaken in local/district hospitals, but in the majority of hospitals, surgeons performed a limited number of procedures (one or two per month). The vast majority of hospitals used two different surgical techniques, open mesh techniques being the preferred technique. Most hospitals made limited use of laparoscopic techniques. Sick-leave periods were shorter after surgery with open mesh techniques as compared to traditional techniques. Less than 20% of hospitals had established control regimens. INTERPRETATION: A clear tendency towards standardization of inguinal hernia repair can be observed throughout the country. Open mesh techniques have now replaced the traditional operative methods, whereas laparoscopic technique has not been adopted. Quality assessment of inguinal hernia repair could be improved. PMID- 11875924 TI - [Collaborated admission to a surgical department]. AB - BACKGROUND: The medical case record is an important tool for securing high quality in treatment and care of patients. Efficient and accurate documentation of diagnostic and therapeutic procedures, patient information etc. is crucial. MATERIAL AND METHODS: We have prospectively studied how time and patient satisfaction are influenced when nurse and doctor together ("combined model") take the patient's medical history, perform a clinical examination, and inform the patient, compared to separate history-taking by nurse and doctor ("separate model"). RESULTS: The mean nurse time per patient increased from 17 min in the separate model to 21 min in the combined model (p = 0.01). Corresponding time use by the doctor was 26 min and 21 min (p = 0.04), and for the patient 43 min and 23 min (p < 0.001). Patients reported the two models to be comparable in quality. INTERPRETATION: The "combined registration model" secures high quality of the medical case record, respects the patient's time, and signals professional collaboration to the patient. PMID- 11875926 TI - [Crohn disease of the oral cavity, illustrated by some cases]. AB - BACKGROUND: We have several times experienced that Crohn's disease has not been diagnosed until several years after the start of oral symptoms. This article presents some patients with oral manifestations of Crohn's disease, as a contribution to more awareness of this possibility. MATERIAL AND METHODS: Clinical and histological data from four patients are presented. RESULTS: All patients had granulomatous inflammation of the oral mucosa. In three patients, the disease started in the mouth. The diagnosis of intestinal Crohn's disease was made from some months up to seven years later. In one patient, no granuloma was found in the intestinal tract, but the patient had intestinal symptoms. Two patients experienced disappointments in their contacts with the local health care system, with considerable delay in diagnosis. INTERPRETATION: We interpret these cases as evidence that a greater awareness of the possibility of Crohn's disease in the oral cavity may lead to a quicker diagnosis, earlier correct treatment, and less patient worry caused by uncertainty. PMID- 11875925 TI - [Lung cancer mortality--now higher in women than in men under 50 years]. AB - BACKGROUND: The smoking prevalence in Norway has evolved differently in men and women. This article examines to what extent this difference is reflected in the lung cancer mortality. MATERIAL AND METHODS: Data on lung cancer deaths and mean population have been taken from official publications from Statistics Norway and grouped in five-year intervals from 1951-55 until 1991-95 and the two-year period 1996-97. Age-adjustments have been made by the direct method against the world standard population. RESULTS: Lung cancer mortality in men peaked in 1991-95, whereas in women it is strongly increasing. In the age group 30-49 years, the mortality in 1996-97 was higher in women than in men, and it is as high as it has ever been in men. If age-specific mortality today had been the same as it was 40 years ago, there would be 81% fewer deaths from lung cancer per year. INTERPRETATION: Women are now paying the price for their smoking habits. This study suggests that smoking is a stronger risk factor for lung cancer in women than in men. PMID- 11875928 TI - [Does supplementation of essential fatty acids influence the visual acuity and cognitive function of term infants?]. AB - BACKGROUND: Norwegian health authorities recommend supplementation of essential fatty acids such as cod liver oil to term infants from four weeks of age. The essential fatty acids are seen as contributing to optimal development of the visual acuity and cognitive functions. This study reviews recent research on this issue. MATERIAL AND METHODS: We assessed six randomized controlled trials, published in 1998 and 1999, of which two evaluated visual function and four cognitive development. Outcome were measured by a number of standardized tests. We compared the results from our review with results from a Cochrane review which evaluated six studies published from 1995 to 1997. RESULTS: Only one of the tests used in the visual acuity studies showed significant effect at three out of the four age-related points in time at which the test was administered. Only one of the tests used in the cognitive development studies showed significant effect. INTERPRETATION: The studies reviewed here are in accordance with previous research and do not provide evidence for the claim that supplementation with essential fatty acids as such is significant in order to achieve optimal visual function and cognitive development in healthy term infants. PMID- 11875927 TI - [Microarray technology--potential in cancer research]. AB - BACKGROUND: Researchers have worked for decades to solve the enigma of cancer. We know that essential checkpoints in the life cycle of cells have to be disrupted in order to create a tumour cell, and some of the genes and proteins involved have been identified. Most of the previous work on identifying these genes have been based on "educated guesswork", as the methods and technologies used have been limited to the examination of genes one by one, or a few at a time. MATERIAL AND METHODS: Microarray technology allows tens of thousands of genes to be examined at the same time, without any previous information on the genes. Both expression levels and copy numbers of the genes can be evaluated. Our studies of breast cancer and bone tumours are presented, as well as examples from the literature. RESULTS AND DISCUSSION: Microarray analyses have been used to produce molecular portraits of breast cancer, malignant melanomas and other cancers. These portraits may help in sub-classification of tumours, in prognosis, and in the general understanding of cancer. For example, studies of gene expression patterns of breast carcinomas, similar with respect to classic prognostic markers (such as ER status, grading and morphology), have identified subgroups of patients that show differences in survival. PMID- 11875929 TI - [Cryoablation--a treatment option for inoperable liver tumors?]. AB - BACKGROUND: Most patients with primary and secondary liver tumours are inoperable by conventional surgery. This has prompted the development of different techniques of local destruction of liver tumours, i.e. cryosurgical ablation radiofrequency, laser, and microwave ablation. MATERIAL AND METHODS: On the basis of relevant literature and our own experience we describe the principles of local destruction by cryoablation of colorectal metastases. RESULTS: Indications for ablation are mainly colorectal metastases and hepatocellular carcinoma. Mechanisms for tumour destruction include intra- and extracellular ice crystal formation, cellular membrane rupture, cellular dehydration and ischaemic damage. Ablation is regularly monitored by ultrasonography, which is suboptimal because of inadequate visualisation of the iceball. Long-term outcome of local destruction of liver tumours is not documented and randomized trials are not ethically acceptable. This complicates analyses of patient outcomes. INTERPRETATION: Local ablation of liver tumours is experimental therapy and should only be performed as a part of prospective trials. PMID- 11875930 TI - [Elective surgery--cancellations, ring fencing and efficiency]. AB - BACKGROUND: We set out to study to what extent emergency admissions disturb elective surgical activities, resulting in decreasing efficiency and fewer elective procedures. We looked at the evidence of whether ring fencing elective surgery is an effective strategy to reduce disturbance from emergency activities, and improve the overall efficiency in hospitals with emergency as well as elective activities. MATERIAL AND METHODS: We systematically searched for published studies that have addressed the above mentioned issues. RESULTS: Approximately 10-17% of elective surgical operations are cancelled. In a Norwegian study, about 14% of cancellations seem to be caused by interference from emergency admissions. We did not find any empirical evidence on the effects of ring fencing elective surgery. INTERPRETATION: The belief that ring fencing is an effective strategy to decrease cancellations and increase hospital efficiency seems to be anchored in a priori reasoning rather than in published evidence. PMID- 11875931 TI - [Caesar Boeck and Boeck's sarcoid]. AB - Caesar Boeck (1845-1917) and his uncle Carl Wilhelm Boeck (1808-1875) established dermatovenerology in the 19th century in Norway. Caesar Boeck began his academic career in the skin department at Rikshospitalet in Oslo in 1875 where the was appointed chief in 1889 and professor in 1895. He retired in 1915. Caesar Boeck was a great clinical capacity, a gifted teacher and a prolific writer. His most important scientific work is the identification of the disease named after him as Sarcoidosis Boeck. PMID- 11875932 TI - [Do nitrates reduce the risk of drug induced hemorrhage of the upper gastrointestinal tract?]. PMID- 11875933 TI - [Humanitarian challenges of a new millennium]. PMID- 11875934 TI - [Sponsorship, authorship and accountability]. PMID- 11875935 TI - [Lamotrigine, documentation requirements and rejection rules]. PMID- 11875936 TI - [The hospital reform and economical profit]. PMID- 11875937 TI - [Specialist education in obstetrics and gynecology]. PMID- 11875938 TI - [Was Hitler just paranoid?]. PMID- 11875939 TI - [Colloidal silver instead of antibiotics]. PMID- 11875940 TI - [Surgical technologies in the treatment of lower extremity varicose disease]. AB - Rational method of surgery for lower extremities varicosis are developed. Method consists of Babkok's extirpation of greater and minor (if it is necessary) saphenous veins with spiral device; preliminary transcutaneous-supracutaneous temporary caprone ligation of changed superficial veins, subcutaneous dissection of ligated veins with minimal blood loss. Original flexible spiral saphenoextractor, inflatable hemostatic device, phlebotoms of simple construction are used. These devices permit to simplify surgery, to decrease tissues trauma and to achieve good cosmetic effect. Recurrence over 1-8 years was seen in 145 operated patients. PMID- 11875941 TI - [Morphologic aspects of transplantation of vascularized tissues complexes]. AB - Microcirculation and tissue leukocytes in transplanted on vascular peduncle tissue complex were studied with light microscopy. Lymph outflow disorders were the causes of transplant edema. Lymph outflow and hemocirculation were restored through new vessels formation that was revealed in all transplant tissues over 2 8 months after transplantation. With time, processes of sclerotic transformation in tissues of transplanted flap progress due to denervation and function loss of transplanted tissues. PMID- 11875942 TI - [Essence and significance of DIC-syndrome in acute peritonitis]. AB - Section specimens of 20 patients who had died of peritonitis of different genesis and specimens of various organs from 10 rats which had of died experimental peritonitis were studied morphologically. It was revealed that 100% patients demonstrated intravascular blood coagulation, it was disseminated in 60% cases and localized--in 40%. In experimental peritonitis intravascular blood coagulation developed in 6 of 10 animals only, and it was disseminated in 3 rats only. It is demonstrated that microthrombi don't lead to focal necrosis due to their special localization in microcirculatory bed vessels. PMID- 11875943 TI - [Differentiated treatment of pulmonary echinococcosis]. AB - Results of operative treatment of 824 patients with pulmonary echinococcosis over the last 25 years were analyzed. In most often encountered oval shaped cysts cappitonage with vertical half-pursestring sutures was the most simple and effective method of residual cavity elimination with the least postoperative complications rate. In 43 patients with cysts of difficult configuration combined method of parasite bed closure with 2-3 surgical methods was used. Differentiated approach to residual cavity elimination has permitted to increase organ-saving surgeries rate from 79.2 to 92.4% that is very important for medical and social rehabilitation of patients. PMID- 11875944 TI - [Plasma technologies in surgery of pulmonary echinococcosis]. AB - 178 patients were operated over 18 years for pulmonary echinococcosis, 100 of them with standard methods, 78--with plasma technologies (surgical device SUPR 2M). Close echinococcectomy was performed in 22.5% patients, open--in 69.1%, combination of these operations--in 8.4% patients. Plasma technologies were used at pneumolysis, cysts section, antiparasitic treatment of residual cavities, fibrous capsule resection (pericystectomy) or cystpericystectomy, marginal resection of the lung with cyst, bronchotomy, sanation of pleural cavity, zone of operation and margins of operative wound. It is demonstrated that postoperative complications were seen in 34% patients operated by standard methods and in 8.9% patients operated with plasma technologies, lethal outcomes were in 2 (2%) and 1 (1.3%) patients, respectively. Plasma technologies permitted to reduce postoperative complications rate 3.8 times and postoperative treatment by 9.8 bed days or by 45.8%. Long-term results were studied in 61 (78.2%) operated patients followed from 1 month to 7 years, there were no recurrence of echinococcosis. PMID- 11875945 TI - [Mediastinal echinococcosis]. AB - Among 29,875 autopsies 59 cases of mediastinal echinococcosis were revealed. Among 4178 patients with thoracic echinococcosis 55 patients had mediastinal echinococcosis. All the patients were operated, most of them underwent ideal ecinococcectomy. Intraoperative prophylaxis of echinococcosis was performed: plearal cavity was treated by low-frequency ultrasound and glycerin. 7 examined patients demonstrated reduction of immune and phagocytosis indices in blood (DC3+, CD4+, CD8+, CD16+, CD21+). Increase of immunoglobulines A, M, G and circulating immune complexes was revealed. Reactions of scolexprecipitation and lymphocytes antigen-fixing were positive. PMID- 11875946 TI - [Immune status in diabetic patients with pyo-necrotic lesions of lower extremities]. AB - Long-term and severe pyonecrotic processes in diabetic patients testify to severe disorders of immune system in this disease. High titer of antibodies to tested autostrain demonstrated its etiologic role in infectious process. The study group consisted of 29 patients (with diabetic pyonecrotic foot lesions), control group- 17 patients with burns of III a, b--IV stage affecting from 20 to 60% of body surface. In diabetic patients antibodies titer to the most encountered infectious agents Staphilococcus aureus and Pseudomonas aeroginosa was lower than in burn patients with immunity deficiency. Decrease of antibodies titer in diabetic patients testifies to high insufficiency of B-immunity. PMID- 11875947 TI - [Ozone-ultrasonic therapy in the treatment of purulent wounds]. AB - Based on planimetric, bacteriologic and histologic study high efficiency of local ozonotherapy of wound in combination with low-frequency ultrasound was demonstrated experimentally on rat model of infected purulent skin wound. This method was used in 45 patients with purulent wounds of soft tissues (postoperative, posttraumatic, burn, sore spot) that led to fast cleaning of wound surface, decrease of bacterial contamination and granulations. It permitted to eliminate inflammation and to create optimum conditions for wound closure. PMID- 11875948 TI - [The method of endo-ventricular repair of the post-infarction left ventricle aneurysm]. AB - Functional importance of left ventricular endoventricular plastic repair in postinfarction aneurysms are discussed. It is proposed to apply vascularized muscle flap consisting of middle and internal myocardial layers as the patch. This policy permits to ensure structural-geometrical optimization of left ventricle and to increase left ventricle contractility due to involvement of automyocardial patch into active contraction. PMID- 11875949 TI - [Central hemodynamics in the remote period in children operated for extrahepatic portal hypertension]. AB - The authors studied functional state of circulation in various terms after operation. Depending on anastomosis creation two group of patients different by visceral hemodynamics change were identified. 58 patients were examined before operation, one year and later after the operation. Functional state of central hemodynamics was evaluated with echo- and Dopplercardiography. It was revealed that after operation the number of children with hyperdynamic type of circulation decreased. A year after distal splenorenal anastomosis creation number of children with hyperdynamic type of central hemodynamics decreased by 20%. Later, regardless of anastomosis type increased cardiac index was revealed in 32.32 and 36.3% children respectively after total and selective anastomosis creation. After total bypass more severe hemodynamic disorders (diastolic dysfunction and inotropic disorders) developed. After distal splenorenal anastomosis creation hemodynamic disorders were seen later due to increase of bypass volume blood flow. Increase of venous return leads to moderate diastolic overloading and myocardial hypertrophy. PMID- 11875950 TI - [Choice of surgical method in patients with inguinal hernias]. AB - Results of surgical treatment of 310 patients with inguinal hernias were analyzed depending on hernioplasty method. Advantages of hernioplasty methods without tissues tension over traditional ones were demonstrated. Indications and contraindications of each method were determined. Algorithm of individual choice of surgery method for inguinal hernias has been developed. PMID- 11875951 TI - [Dispensary-diagnostic department in the system of emergency surgical care]. AB - 3-year experience of dispensary-diagnostic department (DDD) working at the Center of emergency surgical care is analyzed. DDD activity consists of 2 types: dispensary-diagnostic and curative-prophylactic. Department has 5 beds where patients with indeterminate diagnosis are hospitalized (2737 patients over 3 years). Diagnostic beds permit to make diagnosis and select patients with parallel therapeutic procedures. 503 patients were discharged from DDD over 3 years. 534 patients were transferred to surgery department, 97 patients--in internal medicine department. 2% patients required 1-3 hours for final diagnosis, 17%--3-6 hours, 29%--12-24 hours, and only 4%--over twenty-four hours. In 1998- 30% patients didn't require hospital treatment, in 1999--38.5%, in 2000--48.5%. A significant cost-effect was achieved. PMID- 11875953 TI - [Acute appendicitis: disputable problems]. AB - Latest Russian publications about pathogenesis of acute appendicitis, surgical policy and new approaches to its treatment are discussed. Based on many-year experience of surgical treatment of abdominal diseases and literature data, the author gives his opinion about inexpedience of appendectomy for cataral and unchanged appendix. Unfounded proposals towards changed diagnosis formulation in various forms of destructive appendicitis are subjected to criticism. It is concluded that it is necessary to use traditional approaches to diagnosis and treatment of acute appendicitis. PMID- 11875952 TI - [Autologous liver transplantation in localized hepatic lesions]. PMID- 11875954 TI - [Comparative evaluation of topic diagnostic methods of adrenal tumors]. PMID- 11875955 TI - [Congenital additional spleen]. PMID- 11875956 TI - [Lung surgery in N. N. Burdenko surgical clinic of I. M. Sechenov Moscow Medical Academy in 20th century: history]. PMID- 11875957 TI - [What is "clinical evidence" and how to use it]. PMID- 11875958 TI - [Methods of treatment of acute thrombophlebitis of greater saphenous vein]. AB - One-stage combination of thrombectomy on the thigh with compressive sclerotherapy is proposed. This method was used in 66 patients with acute thrombophlebitis of greater saphenous vein (GSV) trunk. It permitted to improve treatment quality, to reduce the scope of subsequent radical operation and to improve its cosmetic result. There were no intraoperative complications. All the patients were discharged in satisfactory state, repeated radical operation for varicose veins of the leg was recommended. PMID- 11875959 TI - Endoscopic retrograde cholangiopancreatography. PMID- 11875960 TI - Starts & facts. Dually eligible patients in the system. PMID- 11875961 TI - Open source powers the medical Internet today and tomorrow. PMID- 11875962 TI - Financial standards for HMOs. AB - In today's health care environment, it is important to assess the liquidity and profitability performance of HMOs. This study focuses on three liquidity ratios and three profitability ratios derived from national databases of between 740 and 776 HMOs from 1996 to 1999. Most of the HMOs appear to be using more debt and are less liquid now than they were in 1995. Since administrative overhead costs and dollars spent on medical costs have been increasing, HMOs' margins have been consistently negative. A more careful analysis of overhead costs and the cost of the delivery of medical services could result in improved HMO quality of care, efficiencies, and a return to positive profit margins. PMID- 11875963 TI - A framework for the practice of pharmacy in the 21st century. PMID- 11875964 TI - Drug approval and pharmaceutical pricing. PMID- 11875965 TI - Can workers' compensation health costs be controlled by managed care? AB - Three recent studies provide, for the first time, the information needed to evaluate objectively the cost savings, utilization, worker satisfaction, and productivity differences of network versus nonnetwork care in workers' compensation cases. The authors review these studies and analyze their findings. PMID- 11875966 TI - How do MCOs use data? It depends on who you ask. PMID- 11875967 TI - A retrospective economic evaluation of olanzapine versus risperidone in the treatment of schizophrenia. AB - This retrospective study evaluates drug treatment patterns and economic outcomes of olanzapine in comparison with risperidone in the treatment of schizophrenia in usual practice. Results showed that patients taking olanzapine versus risperidone stayed on therapy longer (P < .0001) and were prescribed anti-Parkinsonian medications less frequently (P < .005). Compared with risperidone, olanzapine treatment resulted in lower direct mental health care costs ($1,827 less, P < .03) and lower direct total health care costs ($1,834 less, P < .05). The results of this study suggest that the initial selection of an antipsychotic for the treatment of schizophrenia is important: Olanzapine offset its acquisition cost by reducing medical service costs and demonstrated better drug treatment patterns than risperidone. PMID- 11875968 TI - [Kinetics and mechanism of peptide synthesis in solution]. AB - The kinetics of the reaction of Boc-Xaa fluorophenyl esters (where Xaa = Ala, Val, Phe, Ser, Leu, Gly, Met, Pro, or Ile) with leucinamide was studied measuring changes in the fluorescence emission at 375 nm of the fluorophenyl chromophore accompanying the reaction. It was found that the experimental kinetic data couldn't be described by a simple scheme of the second order reaction. The measurements of the kinetic parameters of the reaction at various initial concentrations of reagents indicated that the reaction rate can be expressed as: v = kCNaCAEb, where k is the reaction rate constant, CN is the concentration of leucinamide, and LeuNH2, CAE is the concentration of fluorophenyl ester. The a and b reaction orders were close to 1/2 and 3/2 for Xaa = Ala, Val, Phe, Ser, or Leu, 1/2 and 1 for Gly, Met, or Pro, and 1 and 2 for Ile. The experimental equations for the reaction rate can theoretically be derived from a single scheme of chain reactions with various deactivation ways for active intermediates. The English version of the paper. PMID- 11875969 TI - [The effect of proteolytic removal of the C-terminal fragment of rhodopsin on its ability to activate visual cascade]. AB - The role of the C-terminal domain of rhodopsin in the activation of transducin was studied. The treatment of photoreceptor membranes with trypsin, thermolysin, and Asp-N-endoprotease led to the respective rhodopsin species devoid of 9, 12-, or 19-aa C-terminal fragments. It was shown that the removal of 9-aa fragment by trypsin does not affect the catalytic activity of the receptor, whereas the thermolysin-induced truncation of the rhodopsin C-terminus by 12 aa about 1.5 fold enhances its activity. The Asp-N-endoprotease-assisted removal of 19 aa (i.e., the shortening by seven more C-terminal aa) virtually unchanges the rhodopsin catalytic activity compared to the preparation truncated with thermolysin. These results suggest that the part of the rhodopsin C-terminal fragment between the sites of its cleavage by trypsin and thermolysin (Val337 Ser338-Lys339) inhibits the signal transduction from rhodopsin to the next component of visual cascade. The English version of the paper. PMID- 11875970 TI - [A comparative structure-function analysis and molecular mechanism of action of endonucleases from Serratia marcescens and Physarum polycephalum]. AB - Structural and functional characteristics were compared for wild-type nuclease from Serratia marcescens, which belongs to the family of DNA/RNA nonspecific endonucleases, its mutational forms, and the nuclease I-PpoI from Physarum polycephalum, which is a representative of the Cys-His box-containing subgroup of the superfamily of extremely specific intron-encoded homing DNases. Despite the lack of sequence homology and the overall different topology of the Serratia marcescens and I-PpoI nucleases, their active sites have a remarkable structural similarity. Both of them have a unique magnesium atom in the active site, which is a part of the coordinatively bonded water-magnesium complex involved in their catalytic acts. In the enzyme-substrate complexes, the Mg2+ ion is chelated by an Asp residue, coordinates two oxygen atoms of DNA, and stabilizes the transition state of the phosphate anion and 3'-OH group of the leaving nucleotide. A new mechanism of the phosphodiester bond cleavage, which is common for the Serratia marcescens and I-PpoI nucleases and differs from the known functioning mechanism of the restriction and homing endonucleases, was proposed. It presumes a His residue as a general base for the activation of a non-cluster water molecule at the nucleophilic in line displacement of the 3'-leaving group. A strained metalloenzyme-substrate complex is formed during hydrolysis and relaxes to the initial state after the reaction. The English version of the paper. PMID- 11875971 TI - [A new program complex EFOLD for molecular modeling: the use of a flexible weighting coefficient scheme and the standard valence geometry in modeling the enzyme active sites]. AB - An algorithm for the representation of biopolymer structures in an internal coordinate system (so-called structure regularization) by minimizing the target function with a flexible weighting coefficient scheme using three components that determine the reliability of deviations of each atom was proposed. For the structure regularization, an algorithm for taking into account the temperature factor was suggested for the first time. It was shown by the example of the aspartyl protease rhizopuspepsin that the representation in the internal coordinate system may result in an accurate reproduction of the structural details of separate molecule fragments, such as the active site region of the enzyme. This algorithm was realized as one of the modules of our EFOLD program complex. The English version of the paper. PMID- 11875972 TI - [Coprecipitation of the Pseudomonas fluorescens lipase with hydrophobic compounds as an approach to its immobilization for catalysis in nonaqueous media]. AB - The precipitation of N-cetylamine, N-cetylacetamide, hexan-1,2-diol, cetyl alcohol, and poly(butyl metacrylate) in acetone-water media in the presence of the lipase from Pseudomonas fluorescens was found to be accompanied by the coprecipitation of the enzyme. Within the lyophilized coprecipitates, the lipase exhibits a high catalytic activity and enantioselectivity in the reaction of (1RS)-phenylethanol acetylation with vinyl acetate in t-butyl methyl ether. In order of increasing lipase activity, the coprecipitates can be arranged in the series: cetyl alcohol, poly(butyl metacrylate), hexadecane-1,2-diol, N cetylamine, and N-cetylacetamide, with the activity 2.5- to 19-fold exceeding the activity of the native enzyme. The immobilization of the lipase on solid supports, such as Celite 545 (physical sorption) and Eupergit C250L (covalent binding), in the presence of hexadecane-1,2-diol was found to increase the esterifying activity of the enzyme. The English version of the paper. PMID- 11875973 TI - [Biologically effective sphingolipids devoid of 4-trans-double bond in the sphingoid hydrocarbon chain]. AB - The bioregulatory functions of sphingolipids devoid of 4E-double bond in the sphingoid chain are discussed; the sphingolipids are shown to be biologically active. The English version of the paper. PMID- 11875974 TI - [The lipase/lipoxygenase bienzyme system in AOT reversed micelles in octane]. AB - In this work it is shown that the bienzyme lipase/lipoxygenase system can function in reversed micelles of bis(2-ethyl)hexyl sulfosuccinate (AOT) in octane. As a lipase substrate, a fish fat preparation (fat of sea mammals) with a high content of polyunsaturated fatty acids was used. It was demonstrated that the bienzyme reaction proceeded in a stationary mode and had a rate-limiting step catalyzed by lipase. Under optimal conditions, the efficacy of functioning of the bienzyme system was by an order of magnitude higher than that in water. The lipase/lipoxygenase bienzyme system can be used as a new method of spectrophotometric determination of lipase activity. The English version of the paper. PMID- 11875975 TI - [Modified oligonucleotides containing 1-beta-D-galactopyranosylthymine: synthesis and substrate properties]. AB - A convenient method of regioselective introduction of 1-beta-D galactopyranosylthymine into oligonucleotides was developed and the substrate properties of the modified oligonucleotides were investigated in the enzymic reaction of formation and hydrolysis of internucleotide bonds. The English version of the paper. PMID- 11875976 TI - [Isolation of a highly purified capsular polysaccharide from Salmonella enterica serovar Typhi (Salmonella typhi)--Vi-antigen and its use in serological diagnosis of typhoid fever]. AB - For use in differential diagnostics of typhoid fever, samples of the capsular polysaccharide from Salmonella enterica serovar Typhi (usually named Vi-antigen) were isolated and characterized by physicochemical and serological methods. It was shown that only the sample of Vi-antigen with the minimal (0.57%) admixture of the corresponding lipopolysaccharide (LPS) from S. typhi retained a high serological activity in the tests with monoreceptor anti-Vi sera. However, it exhibited a substantially weaker reaction with sera from normal donors and patients with acute nontyphoid salmonelloses, than Vi-antigen preparations with a higher (0.8-1.2%) LPS content. The chromatographically pure Vi-antigen was purified by triple reprecipitation with hexadecyltrimethylammonium bromide. The content of the LPS admixture in the resulting Vi-antigen samples was quantitatively determined by GC. A high purification level of the Vi-antigen from the LPS admixture allows us to hope that this preparation could serve as a basic component of the test system for the diagnostics of typhoid fever. The English version of the paper. PMID- 11875977 TI - [The synthesis of phosphates of long-chain omega-hydroxyalkyl esters of 11 deoxyprostaglandin E1]. AB - Di(p-methylbenzyl) phosphates of omega-hydroxyalkyl esters of 11 deoxyprostaglandin E1 were synthesized from disubstituted 1,10-decane and 1,22 docosane derivatives for studying permeability of bilayer membranes. The English version of the paper. PMID- 11875978 TI - [Antibiotic N',N''-dibenzyleremomycin with reduced 1,2-peptide bond]. AB - Eremomycin derivatives with benzylated amino groups of both residues of eremosamine and with (R) or (S)-2-amino-4-methylpentyl substituted for N-methyl-D Leu, the first amino acid residue of its heptapeptide, were synthesized to study the role of the peptide bond between the first and the second amino acid residues of the heptapeptide moiety of the antibiotic in its interaction with the precursors of the bacterial cell wall peptidoglycan and exhibition of its antibacterial activity. Comparison of the antibacterial activities of N',N" dibenzyleremomycin, de-(N-methyl-D-Leu)-N',N"-dibenzyleremomycin, and its N-(2 amino-4-methylpentyl)-derivative (1,2-deoxo-N',N"-dibenzyleremomycin) demonstrated that cleavage or replacement of the first amino acid residue by the corresponding aminoalkyl residue results in a decrease in its antibacterial activity towards both vancomycin-sensitive and vancomycin-resistant strains of microorganisms. The English version of the paper. PMID- 11875979 TI - [Nuclease activity of human interleukin-10]. AB - The nuclease activity of human interleukin-10, an immunosuppressive cytokine, was predicted on the basis of structural homology between the 97-105 sequence of human interleukin-10 and the DNA/RNA-hydrolyzing fragment of the endogenous differentiation factor for the HL-60 line of human promyelocyte leukemia cells. The human recombinant interleukin-10 was shown to cleave all forms of plasmid DNA. The role of interleukin-10 in the apoptosis induction in monocytic cells was hypothesized. The English version of the paper. PMID- 11875980 TI - [M.BstF5I-4, the forth DNA methyltransferase of BstF5I restriction-modification system from Bacillus stearothermophilus F5]. AB - The fourth DNA-methyltransferase of the BstF5I restriction-modification (RM) system from Bacillus stearothermophilus F5 (M.BstF5I-4) was discovered, which modifies the adenine residue within the upper strand of the recognition site 5' GGATG-3'/5'-CATCC-3'. Thus, unlike other known RM systems, the BstF5I RM system comprises four genes encoding DNA-methyltransferases, three of which possess the same substrate specificity and methylate adenine within the 5'-GGATG sequence. The English version of the paper. PMID- 11875981 TI - International trauma care: a comparison between Jerusalem, Israel, and Fairfax County, Virginia, USA. AB - BACKGROUND: Trauma is viewed by many as a global problem. The phenomenon of similar outcomes within differing healthcare delivery systems can illuminate the strengths and weaknesses of various trauma systems as well as the effects of these characteristics on patient outcome. OBJECTIVES: To compare and contrast demographic and injury characteristics as well as patient outcomes of two urban/suburban trauma centers, one in Israel and the other in the United States. METHODS: Study data were obtained from the trauma registries of two trauma centers. Demographic variables, injury characteristics and outcomes were compared statistically between registries. RESULTS: Significant differences between the registries were found in demographic variables (age), injury characteristics (Injury Severity Score and mechanism of injury), and outcome (mortality and length of stay). Age and Injury Severity Score were found to be significant predictors of outcome in both registries. The Glasgow Coma Score was found to contribute to patient outcomes more than the ISS. Differences were found in the relative impact of injury and demographic factors on outcomes between the registries. After including the influence of these factors on patient outcomes, significant differences still remained between the outcomes of the trauma centers. CONCLUSIONS: Despite possible explanations for these differences, true comparisons between centers are problematic. PMID- 11875982 TI - Effect of basic fibroblast growth factor on left ventricular geometry in rats subjected to coronary occlusion and reperfusion. AB - BACKGROUND: Previous studies have demonstrated myocardial salvage by basic fibroblast growth factor administration following chronic myocardial ischemia or acute myocardial infarction. OBJECTIVES: To study the effect of bFGF on left ventricular morphometry following coronary occlusion and reperfusion episode in rats. METHODS: bFGF (0.5 mg) or placebo was continuously administered for a period of one week using an implanted osmotic pump. Animals were sacrificed 6 weeks after surgery and myocardial cross-sections were stained with Massontrichrome and with anti-proliferating cell nuclear antigen antibody. RESULTS: LV area, LV cavity diameter, LV cavity/wall thickness ratio, and injury size were unchanged compared with control animals. Proliferating endothelial cells were significantly more abundant in injured compared with normal myocardium, but with no differences between animals treated or not treated with bFGF. CONCLUSIONS: One week of systemic bFGF administration following coronary occlusion and reperfusion had no additional effect on LV geometry or cellular proliferation in rats. PMID- 11875983 TI - Transcatheter ablation of septal hypertrophy: a promising alternative to surgery in hypertrophic obstructive cardiomyopathy. PMID- 11875984 TI - Beware of comparing international health systems, including trauma units. PMID- 11875985 TI - Congenital toxoplasmosis in Israel: to screen or not to screen. PMID- 11875986 TI - The accuracy of diagnosis of Parkinson's disease. PMID- 11875987 TI - Mechanical alternatives to the human heart: paracorporeal assist systems. AB - The currently available paracorporeal assist systems provide reliable short or long-term mechanical assistance to the failing heart, albeit necessitating continuous hospitalization. The intracorporeal assist systems, which provide out of hospital assistance, will be described in the next part of this review. PMID- 11875989 TI - Infant choking or the "A" of resuscitation. PMID- 11875988 TI - Severe combined immunodeficiencies of the common gamma-chain/JAK3 signaling pathway. PMID- 11875990 TI - Unusual simple bone cyst of the distal radius in a toddler. PMID- 11875991 TI - Warfarin therapy in a patient homozygous for the CYP2C9 3 allele. PMID- 11875992 TI - A nerve-racking syncope. PMID- 11875993 TI - Typhlitis: a computed tomography diagnosis. PMID- 11875994 TI - Intratesticular varicocele: diagnosis by power Doppler sonography with the Valsalva maneuver. PMID- 11875995 TI - Clinical learning experiences of Israeli medical students in health promotion and prevention of cancer and cardiovascular diseases. AB - BACKGROUND: The importance of health promotion and disease prevention in health policy and clinical practice is widely accepted in many countries. However, a large number of medical schools do not dedicate a significant part of their curriculum to these aspects. In Israel, there are no reports on the training of the future physician towards his or her role as health promoter in general, or in the areas of cardiovascular and cancer diseases specifically. OBJECTIVES: To examine the preparation of Israeli medical students for the role of health promoter in cancer and cardiovascular diseases. METHODS: The study was carried out over 2 years in two of the four medical schools in Israel: the Sackler Faculty of Medicine at Tel Aviv University and the Faculty of Health Sciences at Ben-Gurion University in Beer Sheva. The students (n = 172, 70% response rate) were surveyed during 1999-2000 by means of a questionnaire, which included assessment of their training towards the role of health promoter, their clinical experiences and exposure to patients at different stages of illnesses at various medical sites, and the specific skills and relevant knowledge they acquired. RESULTS: Most of the students' learning experiences occurred in hospitals with patients at the treatment stage and little time was dedicated to prevention, especially in the community. They demonstrated better knowledge, skills and satisfaction with their learning experiences in CVD than in cancer; and reported having insufficient exposure to several common cancer diseases and lacking examining skills for early detection of cancer. The students in Beer Sheva had significantly more interaction with patients at different stages of CVD and acquired more examination skills than the Tel Aviv students. CONCLUSIONS: A change in the curriculum is urgently needed: namely training medical students in community settings and preparing them to promote the well-being of their patients, including prevention. Attention should be given to launching new learning modes in the pre-clinical and clinical curriculum. We propose that: a) pre-clinical courses include prevention techniques in CVD and cancer, problems of cancer patients, and some examining skills; and b) the clinical phase should integrate oncology concepts and total cancer and CVD care into existing clerkships in the hospitals and in the community. PMID- 11875996 TI - MEDAX 2001--summary of the medical corps session. PMID- 11875997 TI - The notion of health: a conceptual analysis. AB - The notion of health used in medicine may have important implications, such as guiding the allocation of medical resources. This paper explores the notion of health through an overview and conceptual analysis of various notions of health found in modern medical and philosophical literature. It argues that health is characterized either positively or negatively (per exclusion), and either mechanistically (as the set of common or ideal states of a species) or holistically (as unimpaired self-organization of organisms). The paper concludes that a sound notion of health characterizes health negatively and holistically, assimilating mechanism as a good approximation in simple cases. PMID- 11875998 TI - Non-surgical myocardial reduction in hypertrophic obstructive cardiomyopathy. AB - BACKGROUND: Percutaneous transluminal septal ablation was recently introduced as an alternative to surgical treatment of hypertrophic obstructive cardiomyopathy. In this procedure, alcohol is injected into a proximal septal artery to create a localized myocardial infarction. OBJECTIVES: To characterize the immediate and medium-term results following PTSMA. METHODS: Of 13 patients referred for PTSMA, 8 were found suitable for the procedure. Hemodynamic parameters were evaluated prior to and following the procedure, and clinical and echo-Doppler parameters at 2 weeks and 9 months later. RESULTS: The procedure was technically successful in all patients. Resting left ventricular outflow gradient at rest (by Doppler) fell from 82 +/- 37 to 15 +/- 8 mmHg (P < 0.001) 9 months later. Late post-procedural gradient after the Valsalva maneuver was 2 +/- 24 mmHg. The degree of mitral regurgitation fell from 2.0 +/- 0 to 1.5 +/- 0.5 (P < 0.05). New York Heart Association class for dyspnea improved from 2.8 +/- 0.5 to 1.8 +/- 0.8 (P < 0.01) and Canadian Cardiovascular Society class for angina from 2.0 +/- 1.3 to 1.3 +/- 1.2 (P = 0.08). Complete right bundle branch block developed in six patients, temporary complete atrioventricular block in three, and persistent block requiring permanent pacing in one. No flow in the distal left anterior descending coronary artery (presumably due to spilling of alcohol) was seen in one (with development of a small antero-apical infarction) and ventricular fibrillation 2 hours post-procedure in one. None of the patients died. CONCLUSION: PTSMA provided a substantial reduction in left ventricular outflow gradient associated with an improvement in symptomatology. Serious complications are not uncommon. Long-term follow-up is unknown. PMID- 11875999 TI - Laparoscopic versus open appendectomy: results of a retrospective comparison in an Israeli hospital. AB - BACKGROUND: Acute appendicitis is one of the most common conditions requiring surgical intervention. Open appendectomy has been a safe and effective operation for acute appendicitis for more than a century. Recently, several authors proposed that the new technique of laparoscopic appendectomy should be the preferred treatment for acute appendicitis. However, unlike laparoscopic cholecystectomy, LA has not yet gained popularity. OBJECTIVES: To compare open with laparoscopic appendectomy for length of operation, complications, postoperative pain control, length of hospitalization, and hospital costs. METHODS: A sample of 194 patients who underwent OA and LA during 1995 was randomly selected for the study. Patients' demographic data, preoperative laboratory and physical values, histopathologic diagnosis of removed appendix, mean operating time, length of hospitalization, and postoperative pain control and complications were reviewed. RESULTS: Acute appendicitis was confirmed in 66% of patients. The groups were similar demographically (gender and mean age). We could not find any statistical differences in intraoperative and postoperative complications and use of antibiotics. The operative time was longer in the OA group (62.4 vs. 57.3 minutes), but the difference was not statistically significant (P = 0.075). The hospital stay was 2.5 days in the LA group and 2.7 days in the OA group. Higher operative costs were observed in the LA group. CONCLUSION: Laparoscopic appendectomy is comparable to open appendectomy with regard to complications, length of operation, hospital stay, but it is more costly. Laparoscopic appendectomy does not offer any significant benefit over the open approach. PMID- 11876000 TI - Bloom syndrome and Fanconi's anemia: rate and ethnic origin of mutation carriers in Israel. AB - BACKGROUND: The Bloom syndrome gene, BLM, was mapped to 15q26.1 and its product was found to encode a RecQ DNA helicase. The Fanconi's anemia complementation group C gene was mapped to chromosome 9q22.3, but its product function is not sufficiently clear. Both are recessive disorders associated with an elevated predisposition to cancer due to genomic instability. A single predominant mutation of each disorder was reported in Ashkenazi Jews: 2281delATCTGAinsTAGATTC for Bloom syndrome (BLM-ASH) and IVS4 + 4AT for Fanconi's anemia complementation group C. OBJECTIVES: To provide additional verification of the mutation rate of BLM and FACC in unselected Ashkenazi and non-Ashkenazi populations analyzed at the Sheba Medical Center, and to trace the origin of each mutation. METHODS: We used polymerase chain reaction to identify mutations of the relevant genomic fragments, restriction analysis and gel electrophoresis. We then applied the Pronto kit to verify the results in 244 samples and there was an excellent match. RESULTS: A heterozygote frequency of 1:111 for BLM-ASH and 1:92 for FACC was detected in more than 4,000 participants, none of whom reported a family history of the disorders. The Pronto kit confirmed all heterozygotes. Neither of the mutations was detected in 950 anonymous non-Ashkenazi Jews. The distribution pattern of parental origin differed significantly between the two carrier groups, as well as between each one and the general population. CONCLUSIONS: These findings as well as the absence of the mutations in non-Ashkenazi Jews suggest that: a) the mutations originated in the Israelite population that was exiled from Palestine by the Roman Empire in 70 AD and settled in Europe (Ashkenazi), in contrast to those who remained; and b) the difference in origin distribution of the BS and FACC mutations can be explained by either a secondary migration of a subgroup with a subsequent genetic drift, or a separate geographic region of introduction for each mutation. PMID- 11876001 TI - Factors influencing oral colonization in premature infants. AB - BACKGROUND: Factors influencing the oral flora of premature infants have not been adequately investigated. OBJECTIVE: To investigate the effects of gestational age and of anti-bacterial therapy on the oral flora of premature infants. METHODS: Oral cultures were obtained at age 1 day and age 10 days from 65 premature infants, divided into three groups: a) 24 neonates of 30-34 weeks gestation who did not receive ABT, b) 23 neonates of 30-34 weeks gestation who received ABT, and c) 18 neonates < 30 weeks gestation who received ABT. RESULTS: Oral bacterial colonization increased from day 1 to day 10 of life. In 24-34 week neonates, gestational age did not affect early bacteremia or oral colonization at birth. Neither gestational age nor ABT affected late bacteremia or oral colonization at day 10. In 30-34 week neonates with ABT, the oral flora consisted mainly of non Escherichia coli gram-negative bacteria, whereas those who did not receive ABT grew mainly alpha-hemolytic streptococci, Klebsiella pneumoniae and E. coli. In neonates < 30 weeks who received ABT the oral flora were mainly coagulase negative staphylococci. Oral colonization with anaerobes was zero and colonization with fungi was minimal. CONCLUSIONS: Acquisition of oral bacteria rose from day 1 to day 10 of life, regardless of gestational life or ABT. On day 10 of life, the spectrum of oral bacterial flora changed following ABT and consisted mainly of coagulase-negative Staphylococcus and non-E. coli gram negative bacteria. Oral colonization showed few fungi but no anaerobes. These microbiologic observations merit attention when empirical anti-microbial therapy is considered in premature infants suspected of having late-onset sepsis. PMID- 11876002 TI - [X-rays in molecular biophysics: structure-activity relationships and activity of steroid hormones]. PMID- 11876003 TI - Job stress and cardiovascular diseases with health workers. AB - Due to increasing number of patients, reduced possibilities of diagnostics and therapy as well as poor living conditions as a result of total socio-political and economic situation in our country in the last ten years, health workers have been exposed to additional stress factors. Therefore, we wonted to establish to what extent professional stress affects appearance of complications in arterial hypertension with health workers. The examination included employees treated in the health centers "Hospital" and "Novi Sad". According to register for chronic mass non-contagious diseases of MONICA Project, younger patients suffering from arterial hypertension were excluded. From the aspect of possible causative factors of occupation on appearance of cardiovascular complications in patients already suffering from arterial hypertension, the examined group involved doctors and nurses. The control group included laboratory technicians, clerks, cleaners and service workers. It was established that doctors and nurses with hypertension registered in earlier life periods, were more vulnerable to angina pectoris, myocardial infarction and cerebrovascular insult as complications, compared to the rest of employees (if age as a risk factor is excluded) (RR = 3.7; 95% confidence interval 1.6 < RR < 8.6). PMID- 11876004 TI - [Herpes simplex virus and malignancies of female genital organs]. AB - INTRODUCTION: Primary herpes simplex virus (HSV) infections of female genital tract usually end with remission, while the virus remains in the organism--almost in the sacral ganglion in a latent form, protected from humoral and cellular immunity. Stress induces the virus and the result is recurrent genital infection. Frequent exacerbations damage some parts of vital cellular structures without cytolysis, but stimulate malignant transformations. MATERIAL AND METHODS: Vulvar (portio vaginalis uteri) and endometrial tumor tissue samples were analyzed for HSV by direct and indirect fluorescent antibody technique (FAT). Pre and postoperative sera samples were analyzed for presence of anti-HSV antibodies--IgM and IgG by Elisa-Enzygnost method. Acellular filtrates obtained by ultrasonic destruction of malignant tissues were used as inoculum for rabbit corneal scarification. RESULTS AND DISCUSSION: Out of 63 tissue samples, 42 were positive for HSV antigen i.e. 67.3%. According to location 50% of vulvar, 76% PVU and 65% of endometrial tissues were positive. This antigen induces production of virus specific antibodies. Two types of antigens are known: the so-called T-antigen persisting in the cell nucleus and cell-surface antigen--product of the viral genome and can be evidenced by immunofluorescence method. Anti HSV antibodies were present in 63 preoperative serum samples and belonged to IgG group, but not one to IgM, implying a long and chronic course of infection excluding acute primary. Out of 38 postoperative serums the titer of antibodies decreased in 36 evidently, but in two samples remained unchanged. Two samples of endometrial and one from PVU origin contained HSV antigen type one. In the remaining 16 samples HSV 2 antigen was present. Rabbit corneal scarification was the proof of complete infectious virus in malignant tissues. Acellular filtrate of malignant tissues served as inoculum. Corneas of examined rabbits showed a mild inflammation after 24 hours which disappeared in the next 24 hours. We could not isolate the infectious virus by rabbit corneal scarification. Instead of herpetic changes, mild inflammation was evident. This abortive, incomplete symptomatology was probably caused by nonstructural early protein, which is a product of viral genome incorporated in malignant cells. CONCLUSION: On the basis of our results, we can conclude that HSV can have, beside other factors, a very important, maybe an initial role in development of malignant changes of female genital tract, not only on vulva and PVU, but on endometrium as well. HSV I can cause genital infections and have some role in malignant changes as well as HSV 2. However, complete infective virion couldn't be isolated from malignant tissues. PMID- 11876005 TI - [Evaluation of body height and body weight in relation to growth in children in the North Backa region]. AB - INTRODUCTION: Body weight and height in regard to age present an important indicator for evaluation of health status in children. The aim of this study was to evaluate body weight and height in regard to age in children of the North Backa Region (Subotica, Backa Topola and Mali Idos) compared to reference values. MATERIAL AND METHODS: Results of transversal anthropometric measurements performed from 1995-1998 of body weight and body height of 25.790 children aged 1 18 from Health Centers in North Backa Region have been statistically processed by software "CHILD". Body weight and height in regard to age were evaluated by standard deviation, according to international World Health Organization reference values and National Center for Health Statistics (NCHS/WHO). RESULTS: Body weight was established (+2 standard deviation) in 9.33% children, while 1.14% had decreased body weight from -2 standard deviation. Obesity, as well as undernourishment, dominated among children from village Mali Idos. Body height higher than +2 standard deviation was registered in 8.07% children, mostly among boys from Backa Topola. Body height lower than -2 standard deviation was found in 2.10% children of our region. DISCUSSION: Obesity in children aged 1-18, in our region is higher than in the children from UN regions and subregions. Only Russia has worse results generally speaking. In regard to the WHO Global Database of underweight children, we can say that in our children it is hardly present. CONCLUSION: These findings reflect the need for prospective growth monitoring of children in our area and for health improvement through nutrition intervention programs. PMID- 11876006 TI - Typing of field rabies virus strains in FR Yugoslavia by limited sequence analysis and monoclonal antibodies. AB - A total of 32 rabies virus isolates (15 of fox, 14 of cat and 3 of dog origin) from the territory of FR Yugoslavia were collected from December 1996 till February 1998 and analyzed by limited sequencing of N gene and by indirect immunofluorescence and a panel of 20 antinucleocapsid monoclonal antibodies (MAbs). All examined strains were characterized as sylvatic fox strains. Two main genetic variants were detected, 15 isolates belonging to Group I, 14 belonging to Group II, while the remaining 3 could not be classified into any group. This classification was confirmed by MAbs. The obtained results indicate at least two independent cycles of rabies transmission, probably resulting from multiple modes of transmission to the territories now belonging to FR Yugoslavia. PMID- 11876007 TI - [Effect of smoking on lipoprotein metabolism]. AB - INTRODUCTION: Recent aspects concerning effects of smoking on lipoprotein metabolism are presented as well as their harmful effects on blood vessels. It has been proved that smoking stimulates oxidation of LDL particles, producing a significant increase of triglycerides and a decrease of HDL-cholesterol, mostly due to HDL, subfraction diminution. LIPID AND LIPOPROTEIN DISORDERS: Changes of other lipid and lipoprotein parameters are not significant and may even not be found in certain investigations. They are independent of age, body mass, physical activity, alcohol consumption and hormonal status in women; however, they are affected by inappropriate diet. A complete and very fast reversibility of changes after smoking cessation is significant and is important while taking blood for laboratory analyses and interpretation of results. Concerning atherosclerosis, increased oxidation in LDL particles is of great significance. It is established that in vitro exposition of LDL particles to the cigarette smoke induces their increased oxidation and in smokers an increased antibody level of oxidased LDL particles is found. CONCLUSION: A number of arguments speak in favor to the opinion that the unique mechanism by which smoking affects lipoprotein metabolism is its connection to the insulin resistance phenomenon. PMID- 11876008 TI - [Etiopathogenic importance of human herpes viruses type 6, 7 and 8 in manifestations of certain skin diseases]. AB - INTRODUCTION: In the past few years new human herpes viruses (HHV): HHV-6, -7 and -8 have been discovered. According to the most recent literature, they might have an important role in etiopathogenesis of some dermatological diseases. HUMAN HERPESVIRUS 6: HHV-6 was isolated in 1984 from peripheral blood lymphocytes of AIDS patients and patients with different lymphoproliferative diseases. Up to now, two variants of this virus have been identified, A and B, which differ in genetic, biological and immunological characteristics. The etiological importance of variant A, has not yet been clarified, while variant B is considered to be the major cause of many diseases, such as exanthema subitum in infants. In many cases primary infection is associated with elevated temperature, without rash. HUMAN HERPESVIRUS 7: HHV-7 was isolated in 1990 from activated peripheral blood CD4+ T cells of healthy persons. The virus is ubiquitous and more than 80% of babies and infants are affected. Presence of DNA sequences of this virus in mononuclear cells of peripheral blood, skin and plasma of pityriasis rosea patients, points to possible connection between this illness and HHV-7 infection. HUMAN HERPESVIRUS 8: HHV-8 was first identified in tissue samples of patients with Kaposi's sarcoma associated with AIDS in 1994. DNA virus sequences were also isolated in HIV negative persons with Kaposis's sarcoma. Presence of virus can be established in mononuclear cells of peripheral blood, endothelial cells that cover vascular spaces and spindle cells within skin changes. Modes of transmission are still not clarified. However, HHV-8 was identified in some other dermatological diseases as well. PMID- 11876009 TI - [The Wertheim-Meigs radical hysterectomy in surgical treatment of cervical carcinoma]. AB - INTRODUCTION: Wertheim from Vienna was the first to introduce the technique of radical hysterectomy into surgical practice in treatment of uterine cervix carcinoma associated with partial lymphadenectomy (1989). In concern to the level of surgical extensiveness, radical hysterectomy can be divided into 5 classes (after Piver-Rutledge). MATERIAL AND METHODS: At the Department of Obstetrics and Gynaecology in Novi Sad 55 patients with invasive cervical cancer (FIGO st. IB1 IIB) underwent surgery during the period from 1991 to 2001. All patients underwent radical hysterectomy after Wertheim-Meigs technique. The aim of this study was to present operative techniques of radical hysterectomy and complications following surgery. RESULTS: Distribution by stage of the disease was as follows: IB1 = 39 (70.9%), IB2 = 6 (10.9%), IIA = 7 (12.7%) and IIB = 3 (5.5%). On average, there were 15 lymph nodes removed, out of which 9 (16.4%) cases were positive. Postoperative complications were recorded in 19 (34.5%) patients, whereas the incidence of urinary fistulas and intrahospital mortality rate have been reduced to the minimum (0%). Recurrence was evident in 3 (5.4%) patients and 5-year survival rate was 92.3% (in patients operated up to 1997). DISCUSSION AND CONCLUSION: In relation to previously published results of treatment at the Department, there is an evident increase in the number of stage I patients and a decrease in stage II patients, as well as a reduced incidence of postoperative urinary fistulas, total number of complications, but an increase in the 5-year survival rate. PMID- 11876010 TI - [Clinico-epidemiologic characteristics of Lyme disease treated at the Infectious Disease in Novy Sad 1993-1998]. AB - INTRODUCTION: Borrelia burgdorferi, the etiological agent of Lyme disease, is transmitted by the bite of Ixodes ricinus, registered in all parts of Yugoslavia. Vectors are very active in spring and early summer and the disease has a seasonal distribution. Generally speaking, there are three defined stages of the disease, but some of them can be misdiagnosed or really absent. Serological analysis of Lyme disease is very difficult to interpret, especially in later stages, so confirmation by immunoblot assays is recommended. The aim of this study was to present some epidemiologic and clinical characteristics of Lyme disease in Vojvodina in the period from 1993-1998. Throughout this period, 1.659 persons with tick bite were registered, whereas 560 with diagnosed Lyme disease have been treated at the Clinic for Infectious Diseases in Novi Sad. RESULTS: In 511 patients (91.25%) we registered the first stage of the disease, in 42 (7.50%) the second stage and in 7 (1.25%) the third stage of the disease. The mean age of patients with erythema migranes was 38.67 years, mean incubation period was 9.37 days, and tick was removed from the skin after 2.29 days on average. Most of the identified tick bites originated from suburban areas (50.29%), they predominantly occurred in May and June (63.01%), and most of the ticks were removed improperly (57.67%). Dominant clinical manifestations of the second stage were acute meningitis (9.52%), Bannwarth's syndrome (9.52%), arthralgia and arthritis (50%), skin lesions (14.28%), cardiac disorders (11.90%) and mild liver lesions (2.38%) and generalized lymphadenopathy (2.38%). Chronic neuroborreliosis (42.85%), acrodermatitis chronica atrophicans (28.57%) and chronic arthritis were dominant clinical manifestations of the third stage. Up to 81.63% of patients with late stage of disease had a history of previous tick bite. One third of patients were asymptomatic in the first stage of the disease. Improper treatment of the first stage resulted in development of late stage disease in 57.14% of patients. CONCLUSIONS: Morbidity of Lyme disease in Vojvodina is about 1.98-9.8 cases on 100.000 inhabitants, and it belongs to regions with low incidence. Majority of bites are registered during summer months arround cities. Longer persistence of vector on the skin is a risk factor for manifested diseases. Inappropriately treated or completely untreated persons have higher risk for disseminated infection. In our country, most common manifestations of the second stage are neurological and articular manifestations as in the third stage. Leading symptoms and epidemiology aren't enough for diagnosis of Lyme disease, and more specific and sensitive serologic assays are necessary. PMID- 11876012 TI - [Characteristics of morbidity in school-age children in families at social risk]. AB - INTRODUCTION: Social and economic circumstances have a significant influence on children's health. The health status differs from country to country and it depends on gross national product. The aim of the study was to investigate characteristics of morbidity of school children from disadvantaged families. MATERIAL AND METHODS: The study included 232 school children from disadvantaged families who were hospitalized at the Institute of Child and Adolescents Health Care in Novi Sad during a five year period. Disadvantaged families are those with the following risk factors: low income, parents who are not able to comply with parenthood, unsuitable parents, single parent. Discharge diagnoses were analyzed. Diagnoses of children from disadvantaged families were compared with diagnoses of other hospitalized children. RESULTS: School children from disadvantaged families were significantly more often hospitalized because of infectious diseases, parasitic infections, mental disorders, neurologic disorders, symptoms, signs and undefined conditions, poisonings and injuries than children from families without social risks. School children from disadvantaged families significantly more often suffered from mental retardation, emotional and behavior disorders, epilepsy, cerebral palsy and poisonings, than other hospitalized children. CONCLUSION: School children from disadvantaged families have a specific morbidity which differs from the morbidity of children from families without social risks. PMID- 11876011 TI - [Pharmacomodulation of hypersensitivity rhinitis]. AB - INTRODUCTION: Hypersensitive rhinitis is characterized by accumulation of mast cells and eosinophils (believed to be primary effector cells) in the nasal epithelium. Treatment of hypersensitive rhinitis is directed towards reducing either tissue accumulation of these activated cells or the end-organ effects of the released mediators. The aim of this study was to examine effects of local corticosteroid treatment on the number of eosinophils in nasal secretion of patients with isolated hypersensitive rhinitis. MATERIAL AND METHODS: A total of 55 subjects were included in a prospective controlled trial. Thirty-seven patients with hypersensitive rhinitis were included in the experimental group, and 18 patients with hypersensitive rhinitis in the control group. A local corticosteroid, beclomethasone dipropionate, aqueous nasal spray, was administrated every 12 hours (400 micrograms per day) to the experimental group during 6 weeks. Patients of both groups were regularly controlled during the study period by ENT and cytological examination of nasal secretions. RESULTS AND DISCUSSION: Very significant differences in regard to the number of eosinophils were found in the nasal secretion of experimental patients with hypersensitive rhinitis, whereas they were insignificant in patients from the control group. Strong antieosinophilic effect of beclomethasone was evident. Eosinophil apoptosis at the inflammatory site appeared to be delayed when interleukin-5 was generated by neighbouring cells, or this delay is due to autocrine production of this cytokine. CONCLUSION: Local corticosteroid treatment of hypersensitive form of chronic rhinitis significantly reduces the number of eosinophils and thus probably modulates the pathogenesis of this inflammatory process. PMID- 11876013 TI - [Neurotrichinosis]. AB - INTRODUCTION: Trichinellosis is an anthropozoonosis caused by Trichinella spiralis. Central nervous system complications can occur during the course of this disease. This paper presents a case of neurotrichinellosis, assayed by using indirect immunofluorescence test. CASE REVIEW: A patient aged 30, was admitted to the Clinic on the fifth day after onset of the disease, which was gradual, starting with flu-like symptoms. Two days before admittance, the diseased became somnolent, disconcerted and disoriented. On admission, the patient was highly febrile (39 degrees C), dehydrated, with eyelid edema. Right hemiparesis was present. The disease was confirmed by indirect immunofluorescence test, showing an increase of trichinella antibody titre (1:20, 1:160 and 1:640). The cerebrospinal fluid was cytobiochemically normal. Electroencephalographic findings exhibited a moderate cerebral dysfunction. Multifocal unspecific changes were established by magnetic tomography. The diseased was treated by mebendazole and prednisolone. The course of the disease was favourable and the patient was cured without sequelae. DISCUSSION: Recognition and diagnosis of Trichinellosis are complicated due to its polymorphid symptomatology. In the case reviewed, the disease started with clinical features of flu-like symptoms and febrile gastroenteritis. According to some authors, central nervous system manifestations occurred in 10-15% of the diseased. CONCLUSION: When examining clinically manifested encephalitis, Trichinelosis should be taken into consideration as a cause of the disease. PMID- 11876014 TI - [Atrophic pilar keratosis of the face: case report]. AB - INTRODUCTION: Keratosis pilaris atrophicans faciei (KPAF), previously called ulerythema ophryogenes, belongs to a group of follicular syndromes with inflammation and atrophy. The disease often starts at birth or during the first months of life with autosomal dominant inheritance. CASE REPORT: We report a case of a 24-year-old woman, who noticed the first lesion two years ago. Skin lesion spread symmetrically on the cheeks, forehead and chin. Keratotic follicular papules were surrounded by erythema. After disappearance of follicular papules, atrophy occurred. Histopathological analysis from skin biopsy specimens confirmed the diagnosis of KPAF. DISCUSSION: Follicular syndrome with inflammation and atrophy starts in early childhood, but first lesions can also appear among teenagers or in adults. The course of the disease is progressive with permanent follicular destruction. The histopathologic changes are dynamic and follow the clinical course of the disease. A topical retinoid: tretinoin therapy was introduced. After one month of treatment good effects were visible, including decrease of erythema and follicular hyperkeratosis. PMID- 11876015 TI - [Acquired B antigen in a pregnant woman belonging to the A1 blood group: case report]. AB - INTRODUCTION: This article presents a case of acquired B antigen in a 24 year old pregnant woman, with A1 Rh D positive blood group verified by expression ABO tube test, secretor status, and investigation of group specific substance in saliva, by agglutinating inhibition test and examination of blood groups in patient's family (both parents and sister). CASE REPORT: Since the patient was a clinically healthy woman, without anamnestic data and clinical signs of any kind of disease (such as diseases of the gastrointestinal system, especially colonic disorders), and with regular course of pregnancy, this acquired irregular agglutination of red blood cells, suggested a possible infection with Gram-negative bacteria. That is why she underwent additional investigations of blood and urine. RESULTS AND CONCLUSION: Hemocultures were negative, but urine tests revealed a urinary infection with uropathogenic strains of Escherichia coli O86. Eight months later the patient was retested, and findings of acquired B antigen and latent urinary infection with E. coli still persisted, proving the cause of irregular agglutination. PMID- 11876016 TI - Listening with the creative ear. PMID- 11876017 TI - Racism, vicious circles, and the psychoanalytic vision. PMID- 11876018 TI - Freud's alleged repudiation of the seduction theory revisited: facts and fallacies. PMID- 11876019 TI - What if there were no ghosts: a dissociative perspective on The sixth sense. PMID- 11876020 TI - Determination of carbamazepine and its metabolite carbamazepine-10,11-epoxide in serum with gas-chromatography mass spectrometry. AB - Carbamazepine, one of the most often used antiepileptic drugs, undergoes enzyme biotransformation through epoxidation with the formation of its metabolite, carbamazepine-10,11-epoxide (carbamazepine epoxide). The determination of carbamazepine epoxide is clinically significant in therapeutic drug monitoring as it decreases the risk of toxic reactions and increases the possibility of reaching the expected therapeutic result. The aim of this study was to introduce a gas chromatographic method with mass spectrometric detection to simultaneously determine the serum levels of carbamazepine and carbamazepine epoxide. Blood samples from 80 epileptic patients aged between 1 and 63 years were analyzed. All patients were taking carbamazepine as monotherapy and had achieved the steady state serum drug concentration. A microcolumn extraction of carbamazepine and carbamazepine epoxide was obtained by elution. A gas chromatograph with a mass spectrometer and a 25 m x 0.2 mm ID, 0.33 micron film thickness, crosslinked 5% phenyl-methylsilicone capillary column HP-5 was used. The coefficients of correlation for the calibration plots obtained for carbamazepine epoxide and carbamazepine were 0.988 and 0.995, respectively. The precision, tested for n = 20, showed coefficients of variation within one day as follows: carbamazepine epoxide: 11.0% and 6.55%; carbamazepine: 11.8% and 5.4%. The coefficients of variation from day to day were: carbamazepine epoxide: 9.4% and 6.83%; carbamazepine: 8.5% and 5.7%. The detection limit was 10 micrograms/l and the recovery 82.5% for carbamazepine epoxide and 92.5% for carbamazepine. A fast and simple gas chromatographic method for the routine therapeutic monitoring of carbamazepine and carbamazepine epoxide was developed. The simultaneous determination of the serum levels of carbamazepine epoxide and carbamazepine offers the possibility of measuring the total drug concentration, as well as that of its metabolite, while considering the results of clinical response and the special features of carbamazepine's enzymatic biotransformation through epoxidation with the formation of its metabolite. PMID- 11876021 TI - Biotinylation of antisense oligonucleotides does not alter lipofectin enhanced cellular uptake in prostate cancer cell lines. AB - Biotinylation is a common modification made to pharmaceuticals, including antisense oligonucleotides (oligos), to enhance their specific delivery. Such agents bind to targets that have been previously labeled with conjugated avidin, or alternatively, heteroconjugate monoclonal antibodies that have dual biotin and tumor-specific antigen specificities may be employed. However, for a drug to be efficacious it must also be taken up by the targeted cells. This is frequently difficult for large molecular weight compounds and cationic lipids, like lipofectin, are often employed. However, the effect of biotinylation on oligo uptake has not been examined in the presence of lipofectin, particularly in prostate cancer cells. Oligos conjugated with biotin and FITC were incubated in vitro with LNCaP and PC-3 cells in the presence of a previously determined effective concentration of lipofectin. Fluorescent uptake and distribution was compared to similar oligos that were not biotinylated. The results demonstrate that biotinylation does not alter the uptake of oligos in LNCaP or PC-3 prostate cancer cells, nor does it alter their retention or cytoplasmic distribution in PC 3 cells when used with lipofectin. PMID- 11876022 TI - Effect of repeated administration of the antioxidant stobadine on the behavior of singly-housed male mice in intraspecies conflict. AB - The aim of the present study was to assess the effect of repeated oral administration of stobadine (70 mg/kg) on the occurrence of selected behavioral elements during exposure to an intraspecies conflict between singly-housed and group-housed male mice. Isolation induced timidity (defensive-escape behavior without attacks) in most mice (87%). This isolation-induced timidity was reduced after stobadine treatment. In the stobadine-treated group, sociable activities (following, climbing) were also decreased. After discontinuation of the treatment (18 days), aggressive behavior tended to increase in the stobadine-treated group. The results of this study are indicative of an inhibitory effect of repeated administration of stobadine on some behavioral activities of singly-housed male mice in an intraspecies conflict. PMID- 11876023 TI - Role of nitric oxide in experimental models of psychosis in rats. AB - The aim of our study was to investigate the effects of the NO precursor L arginine and the nitric oxide synthase inhibitor N omega-nitro-L-arginine (L NORAG) on amphetamine-induced stereotypy, haloperidol-induced catalepsy and conditioned avoidance response (CAR) in rats. Amphetamine (3 mg/kg i.p.) was used for the induction of stereotypy, while for the induction of catalepsy and CAR, haloperidol (2 mg/kg i.p.) was used. This study was divided into 2 parts--acute administration of L-arginine (150 mg/kg i.p.) and L-NOARG (50 mg/kg i.p.) and chronic administration of L-arginine (150 mg/kg/day i.p.) and L-NOARG (50 mg/kglday i.p.) for 5 days. We found that L-arginine inhibited amphetamine induced stereotypy and haloperidol-induced catalepsy, but intensified CAR. On the other hand, L-NOARG intensified stereotypy and catalepsy but inhibited CAR. Also, there was no significant difference between the scores of acute and chronic administration of L-arginine and L-NOARG. It is concluded from our study that nitric oxide produces conflicting results on various models of psychosis. L arginine might be useful as an antipsychotic without causing extrapyramidal symptoms. PMID- 11876024 TI - Reversal of pyrogallol-induced delay in gastric emptying in rats by ginger (Zingiber officinale). AB - The effects of the acetone extract of ginger (Zingiber officinale) was studied against pyrogallol-induced delay in gastric emptying in rats. Wistar rats of either sex, weighing between 200-250 g, were used. Pyrogallol, at a dose of 100 mg/kg i.p., significantly delayed the gastric emptying of a methyl cellulose meal. One-hour pretreatment with ginger acetone extracts (100, 250 and 500 mg/kg p.o.) reversed the pyrogallol-induced delay in gastric emptying. The effect was significant at doses of 250 and 500 mg/kg. When the low dose of ginger (100 mg/kg p.o.) was given with vitamin C and vitamin E (100 mg/kg p.o., each), the reversal of gastric emptying was more pronounced than when only two vitamins or ginger (100 mg/kg and 500 mg/kg) were given alone. The present study indicates the potential of ginger in improving symptoms such as abdominal discomfort and bloating, which may accompany several gastrointestinal illnesses. PMID- 11876025 TI - Safety and efficacy of low-dose amphotericin B lipid complex for empirical antifungal therapy of neutropenic fever in patients with hematologic malignancies. AB - Amphotericin B lipid complex (ABLC) has been investigated as an empirical antifungal treatment for neutropenic patients with persistent fever of unknown origin (FUO). We studied the safety and efficacy of low dose ABLC (1 mg/kg/day) for empirical treatment of neutropenic FUO. Sixty-one patients with hematologic malignancies developing 69 episodes of neutropenic FUO after chemotherapy or hematopoietic stem cell transplantation were included in the study. The median patient age was 47 years (18-68). The median duration of neutropenia (< 0.5 x 10(9)/l) was 17 days (7-45) and the median duration of ABLC therapy was 8 days (2 19). Thirteen patients (19%) suffered from mild to moderate infusion-related adverse events. Creatinine levels were stable in 42 cases (61%), improved in 9 (13%) and deteriorated in 18 (26%), with no other significant toxicities. Among 67 evaluable episodes, the response rate (resolution of fever during the period of neutropenia without developing a fungal infection) was 67%, while 33% were treatment failures. Low-dose ABLC is safe, well tolerated and seems to be at least as effective as c-AmB for empirical antifungal therapy of FUO. Randomized trials at this dose level comparing ABLC with c-AmB or other lipid formulations are warranted. PMID- 11876026 TI - DNA research in forensic dentistry. AB - DNA analysis has recently been introduced to forensic dentistry and is now frequently used in identifying individuals or determining the origin of certain tissues. This review reports on teeth and saliva as a source of DNA. Not only the quantity of DNA available for the laboratory is important, but also the quality and purity. Teeth are resistant against extreme circumstances such as temperature, humidity and acidity, which is an important advantage in DNA analysis. Furthermore, an abundance of DNA can be extracted from teeth. Saliva can be obtained in a simple, painless and non-radical way. The double swab method is very effective; DNA recovery is significantly higher with the double swab method compared to the single swab or filter paper method. This review reports on the different techniques used to extract DNA from teeth and saliva, as well as DNA analysis of these samples. The usefulness and advantages of the double swab method for saliva, cryogenic grinding for teeth and the chelex extraction and polymerase chain reaction for both types of samples is also described. DNA analysis has proven its value in forensic dentistry, but ethical and juridical considerations are still a matter of debate and criticism. PMID- 11876027 TI - Overview of the effects of Ringer's acetate solution and a new concept: renal ketogenesis during hepatic inflow occlusion. AB - Developments in infusion therapy with sodium lactate administered with its isomer, and developments in acetate utilization, both as an infusate or a dialysate, have been previously described, as have the various effects of sodium acetate on cardiovascular function, metabolism and fluid replacement. Unfavorable effects of acetate infusate or dialysate, such as cardiovascular depression, have also been reported, and although the number of studies reporting favorable effects on cardiovascular metabolism is increasing, it is still a matter of controversy. It is clear, however, that acetate can be metabolized in the liver as well as in extrahepatic organs yielding bicarbonate, which can be utilized as an alkalizing agent. Recently, the authors reported new experimental findings that sodium acetate was metabolized to ketone bodies by the kidney during hepatic inflow occlusion. Ketone bodies attract considerable attention as energy substrates in the stress phase and in the reduced redox state. Administration of sodium acetate under such conditions may therefore contribute as substrate supply. PMID- 11876028 TI - [Hypoglycemia caused by oral hypoglycemic agents: risk of relapse after normalisation of blood glucose]. AB - Three patients, two women aged 83 and 79 years, and one man aged 61 years, known with diabetes mellitus and using oral blood glucose-lowering drugs, presented with impaired consciousness due to hypoglycaemia. Two of them were admitted and recovered well after recurrence of hypoglycaemia in one and discontinuation of medication in both. The third patient had prolonged hypoglycaemia and died three weeks later, still unconscious, due to pneumonia. Older patients with hypoglycaemia caused by longer acting sulphonylurea derivatives should be admitted, especially if they live alone or have risk factors such as renal or hepatic insufficiency. Intravenous glucose should be given if necessary. Discontinuation of the longer acting sulphonylurea derivative or replacement with a shorter acting drug should be considered. PMID- 11876029 TI - [The Dutch College of General Practitioners' "Gout" Standard: a response from general practitioners]. AB - Although gout has a long nosological history, there are still many uncertainties regarding its pathophysiology, causative factors and most common therapies. Therefore, composing an evidence-based guideline on gout is a challenge. There is a lack of good clinical research, especially in primary care populations where most gout patients are diagnosed and treated. Far more insight is required into the mechanisms which underlie increasing and decreasing serum uric acid levels which, via the blood-synovium barrier, should increase or decrease urate crystals with inflammatory potency. In view of this lack of information, it would have been more appropriate for the Standard not to contain unproven facts and therapeutic recommendations. Guidelines should be kept simple until good clinical research proves the opposite. PMID- 11876030 TI - [The Dutch College of General Practitioners' "Gout" Standard: a response from rheumatologists]. AB - The authors (rheumatologists) agree with most recommendations of the Dutch College of General Practitioners' guidelines for the diagnosis and management of gout. The most important criticism concerns the supposed diagnostic value of serum urate levels: close examination of the data suggests that this value is very limited. PMID- 11876031 TI - [World Health Organization places psychiatry high on the agenda, also consequences for the Netherlands]. AB - In October 2001 the World Health Organization (WHO) devoted its yearly World Health Report to psychiatry under the title 'Mental health: new understanding, new hope'. Psychiatric disorders have a high prevalence and an enormous impact. There is a great need for medical care, rehabilitation and support from family and community. Effective treatments have become available for most disorders. Recommendations for the Netherlands are: to increase care in the community, to better educate the public, to improve national policies, programmes and legislation, to further develop human resources (currently an enormous problem, hindering provision of minimal care for chronically hospitalised patients) and to link mental health more effectively with other sectors in the community (to overcome the vast numbers of people on disability benefit because of psychiatric disorders). PMID- 11876032 TI - [Pregnancy and medical irradiation; summary and conclusions from the International Commission on Radiological Protection, Publication 84]. AB - Prenatal exposure to ionising radiation as used during most diagnostic procedures generally presents no increased risk of prenatal death, malformation or impairment of mental development (i.e. deterministic effects) compared to the background incidence of these entities. Higher doses of radiation used in therapeutic procedures can result in significant foetal harm. In general, malformations only occur above a threshold dose of 100-200 mGy. These doses are not normally reached with most properly executed diagnostic procedures. During the period from 8 to 25 weeks after conception, the central nervous system is particularly sensitive to radiation. Foetal doses in excess of about 100 mGy may result in a decrease in IQ. Between 8-15 weeks after conception, a foetal dose of 1000 mGy (1 Gy) reduces IQ by about 30 points. This reduction is less marked during the period from 16-25 weeks. At foetal doses of 1000 mGy in the period from 8 to 15 weeks after conception the risk of severe mental retardation is about 40%. During the period from 16 to 25 weeks, this risk is practically zero at a dose of 1000 mGy. Radiation exposure of the embryo/foetus is associated with an increased risk of tumour induction (stochastic effect). Recent absolute risk estimates for fatal cancer risk for ages 0-15 year after in utero irradiation have been estimated to be 6% per Gy (0.06% per 10 mGy). For the whole life span this risk is about 15% per Gy (0.15% per 10 mGy). Pre-conception irradiation of either parent's gonads has not been shown to result in increased cancer or malformations in the children. PMID- 11876033 TI - [Errors in methodology (conclusion). XXXIX. The ultimate truth]. AB - Studies that have methodological shortcomings may still lead to the right conclusion. Sometimes, the bias is small, or the shortcoming is not relevant for the issue being studied. It is also possible that a perfectly designed and executed study does not yield the 'right' answer. For example, chance circumstances might lead to the incomparability of groups. Bayes' theorem clarifies why different experts may come to opposite conclusions based on the same studies. The ultimate truth remains elusive. PMID- 11876034 TI - [Summary of the Dutch College of General Practitioners' "Gout" Standard]. AB - The typical form of acute gout can be clinically diagnosed. The term 'complicated gout' is used if there are more than three acute attacks of gout per year, tophi or urate stones in the urinary tracts. In the case of recurrent probable acute gout, a diagnostic fine needle aspirate from the joint during an attack is indicated. First choice treatment of acute gout consists of NSAIDs. Colchicine is the second choice treatment and the third choice treatment consists of corticosteroids. Excessive alcohol use should be limited. Treatment of chronic gout depends on the uric acid excretion in the 24-hour urine. If the level of excretion is too low, the first choice should be benzbromarone, and if the uric acid output is too high, allopurinol should be the treatment of first choice. Increased fluid intake is recommended; maintenance treatment with colchicine is not advised. Consultation with or referral to a rheumatologist is indicated in the case of doubt about the diagnosis of 'acute gout' or 'complicated gout', or (suspected) bacterial arthritis and insufficient treatment effect. PMID- 11876035 TI - [Diagnostic image (76). A woman with acute pancreatitis]. AB - A 59-year-old woman had severe acute pancreatitis and Grey Turner's sign of bluish discoloration of the flanks with local infiltration at CT. PMID- 11876036 TI - [Reported cases of Legionella pneumonia in the Netherlands, 1987-2000]. AB - OBJECTIVE: To describe the incidence and mortality of legionnaires' disease (LD) in the Netherlands on the basis of registration, with a focus on possible sources of infection, and geographic and seasonal variations. DESIGN: Retrospective, descriptive. METHOD: The incidence and mortality rate of LD from July 1987 until December 2000 was described using registration data from the Dutch Health Inspectorate. Possible sources of infection were also listed (patients involved in the Bovenkarspel outbreak (1999) were excluded). Geographic variations in occurrence of LD were investigated by incidences computed per province. To gain insight into the possible existence of seasonal variations the mean numbers of patients per month were compared. RESULTS: During the period of interest a total of 806 persons with LD were reported, 36 of whom were excluded. The mean incidence rose from 0.27 cases per 100,000 inhabitants in the 1987-1998 period to 0.91 cases per 100,000 inhabitants in the 1999-2000 period. The male:female ratio was 2.6:1. A total of 96 persons (12.5%) died as a result of LD. There were more deaths in the group of patients with confirmed LD compared to the group of patients with probable LD. After 1998 the case fatality rate was lower than in the previous period (Fisher's exact test: p = 0.005). The potential source of infection was situated in the open population in 92.9% of cases. In 7.1% of cases the potential source was situated in a hospital. The incidence per province varied from 0.13 per 100,000 (Drente) to 0.66 per 100,000 (Limburg). During the summer months more patients were reported, most of whom had contracted the infection abroad. In contrast to the 1987-1998 period, after 1998 the incidence in the Netherlands was higher than the European mean and the incidence of confirmed LD was higher than in the United States. PMID- 11876037 TI - [A man with plasma cell dyscrasia and polyneuropathy due to POEMS syndrome]. AB - In a 52-year-old man with general malaise, muscle stiffness and weakness, POEMS syndrome was diagnosed based on polyneuropathy, splenomegaly, lymphadenopathy, subclinical hypothyroidism and the presence of a monoclonal paraprotein with osteosclerotic lesions and an indurated skin (POEMS is an acronym for Polyneuropathy, Organomegaly, Endocrinopathy, Monoclonal protein, Skin changes). This is a rare systemic disease from the clinical spectrum of plasma cell dyscrasias with polyneuropathy. The clinical picture is broader and more pleomorphic than the acronym suggests. The possibility of a POEMS syndrome should be considered in the differential diagnosis of polyneuropathy in association with monoclonal gammopathy. Quite often it is associated with osteosclerotic myeloma or mixed osteoscleroticlytic lesions. The patient described was treated with high dose corticosteroids which were gradually decreased over the next three months, upon which a marked improvement could be seen. The general malaise subsided, as did the splenomegaly, and the skin became supple again. PMID- 11876038 TI - [Gas configuration between the liver and the right diaphragmatic dome: Chilaiditi's sign]. AB - A 64-year-old man of Hindustan origin presented at the Emergency Department with symptoms which suggested stomach perforation. A chest X-ray taken with the patient in the upright position showed gas between the liver and the right hemidiaphragm. After 30 minutes the pain subsided spontaneously. The gas seen on the chest X-ray was diagnosed as Chilaiditi's sign, caused by interposition of the colon between the liver and the diaphragm. This interposition is usually asymptomatic and does not require surgical treatment. PMID- 11876039 TI - [Marius Tausk (1902-1990), influential endocrinologist and producer of medicines; a retrospect to mark the centenary of his birth]. AB - Marius Tausk, born in Sarajevo, studied medicine at Graz in Austria. In 1926, he attended a socialist youth rally in Amsterdam. Whilst there, he met Prof. Ernst Laqueur, a pharmacologist in Amsterdam, who offered him a position at Organon, a newly-founded pharmaceutical company in Oss, the Netherlands. He remained in the Netherlands and became the driving force behind this innovative company. Tausk made many contributions to new developments across the field of endocrinology, including the discovery of the adrenal steroids (together with T. Reichstein, Nobel Prize Laureate in 1950), and the development of oral contraceptive pills. He was astute enough to quickly patent the first corticosteroids. He could quickly extract the essential elements from an information source and disseminate this in five languages. His sharp judgment brought him friends and admirers, yet also those who feared him. In 1937 he was appointed as a private lecturer at the Medical Faculty of Utrecht University and in 1956 he was made Special Professor of Theoretical Endocrinology. He was awarded a number of scientific distinctions including two honorary doctorates. In 1949 he was honoured with the Knighthood of the Order of The Netherlands Lion. PMID- 11876040 TI - [Medical research and the pharmaceutical industry. Uneasy lovers or premarital agreements?]. PMID- 11876041 TI - [Development of hearing aids]. PMID- 11876042 TI - [The hospital psychiatrist as consultant in assisted suicide requests: clinical experiences in a oncology center]. PMID- 11876043 TI - [External quality assurance of medical rehabilitation in statutory health insurance]. AB - Medical rehabilitation and the statutory health insurance companies (SHI) are increasingly coming into conflict due, on the one hand, to the current shortage of resources in the health care system and, on the other hand, the increasing need for rehabilitation measures. This makes it necessary to base the planning and management of rehabilitation treatments on economic and qualitative considerations equally. Quality assurance procedures play an important role in guaranteeing that rehabilitation is effective and efficient. This article discusses the expectations of the SHI companies regarding quality assurance (QS), explains the demands made by the legislator and reports on the activities of the SHI companies aimed at implementing a common external QS-program. As rehabilitation units are treating patients for various health insurance companies it is imperative that these agencies--in spite of competing with each other- agree on a common QS-program in order to avoid a situation in which a rehabilitation unit would have to provide different (and perhaps conflicting) proofs of its quality. PMID- 11876044 TI - [Quality management in medical rehabilitation--a comparison of national and international approaches]. AB - The paper presents an integrated quality management program that is being run in acute-care hospitals and rehab facilities in Germany and in the U.S.. The experiences gained through all commonly implemented quality management concepts have been incorporated; which has allowed the development of comparison and integration tools. The overall experience indicates that the contents of good quality management are fairly independent of the concept applied and uniform as to their core. By using an integrated Quality Report, accreditation can be achieved while striving for excellence. The experiences gained through the KTQ Pilot Phase, the QMK Field Phase, and Joint Commission applications in Germany as well as in the hospitals of Pacific Health in the U.S. were incorporated. The sum total of experiences with more than 100 projects indicates that it is easy to combine two or more assessment models in such a manner as to reap the benefits of the models without incurring the expense of the extra work involved. The author recommends that German rehab facilities engage in quality assessment as soon as possible. Owing to insufficient clinical parameters in Baldrige and EFQM, crosswalk complementation with clinical evaluation instruments such as Joint Commission or KTQ and QMK is advisable. PMID- 11876045 TI - [Treatment goals as an instrument of quality management in psychosomatic rehabilitation]. AB - The present article outlines the development of a system of categories of treatment goals in psychosomatic rehabilitation. As a first step a content analysis of 242 letters of discharge was carried out. In order to give a systematic describe of the thus extracted goals a system of categories was developed. Which was then in a second step used as the basis for detailed discussions by psychosomatic rehabilitation experts aimed at revising and optimizing the system of categories. This developed classification of treatment goals--owning to the possibility in principle of operationalization--will provide the conditions for further strategies within the context of evaluation and measures of quality assurance. PMID- 11876046 TI - [Indicators of process and outcome quality in the context of multicenter quality management in cardiac rehabilitation]. AB - From the point of view of quality management in general and clinic comparisons in particular, indicators of outcome quality must be chosen in such a way that conclusions regarding the quality of rehabilitational care processes can be drawn. Moreover, these indicators should reflect disease characteristics which, on the one hand, are relevant in terms of prognoses and, on the other, provide indications for rehabilitation therapy. Predictors of rehabilitation success need to be differentiated analytically in order to safeguard the adjustment of risks when, for instance, clinics are being compared. Suitable outcome indicators for cardiological rehabilitation are LDL-cholesterol and functional-physiological capacities. Taking low-density lipoprotein (LDL) as an example, a well-tested approach to applying the results of studies, in this case those of the Cardiac Rehabilitation Outcome (CARO) study, to internal and external quality management is shown. The way in which information is provided by a quality information system is explained. The scheme is to be expanded into a routine monitoring system with the aid of a nation-wide follow-up study (CARO-II). The studies are being supported by the DGPR cardiological society. PMID- 11876048 TI - [The nationwide quality assurance program of the German pension insurance scheme. Experiences of several years of routine implementation]. AB - The German pension insurance scheme's quality assurance programme for medical rehabilitation is aimed at improving the quality of medical rehabilitation. Interviews with patients and peer-review of medical files show that patient satisfaction is high and that experienced staff perceive the quality of the rehabilitation processes to be high. The results of external quality assurance procedures have also helped to improve the quality of rehabilitation centres. PMID- 11876047 TI - [An outcome-oriented quality management strategy for the inpatient rehabilitation of orthopedic patients--towards an empirically based "best practice"]. AB - This paper presents the development and testing of an outcome-oriented quality management system for the inpatient rehabilitation of orthopaedic patients, with a special focus on the indication of knee-replacement. On the level of outcomes we describe the development of appropriate indication-specific outcome indicators, capable of being used to rank the performance of hospitals. With regard to the process-quality of the empirical analysis of the rehabilitative therapeutic services possibilities and limitations are shown. Furthermore an inkling of the shape that an expert-consent-based base of evidence for identifying best practice--arrived at by using primary empirical findings--might assume is presented. The procedure for establishing the latter offers a methodical approach toward continuous quality development of orthopaedic inpatient rehabilitation, one that is characterised by guidelines and performance transparency. Development potentials are recognised in the formal and the content quality of the therapeutic services. PMID- 11876049 TI - [Quality assessment by patients as a method for securing the quality of the rehabilitation of persons addicted to alcohol]. AB - We present a discharge questionnaire given to patients at the end of their inpatient treatment for alcohol addiction. The questionnaire poses questions regarding the satisfaction of the patients with the therapy received, the improvements in their state of health and allows them to rate the rehabilitative strategy and the therapeutic programme. The results show a very close correlation in the light of behavioural medicine between the rehabilitative strategy and the improvement rates. PMID- 11876050 TI - [Outcomes count: the importance of measuring the outcomes of hospital care]. AB - A century ago the Boston surgeon Earnest A. Codman described in detail the requirements for monitoring quality of care and emphasized the importance of outcomes in evaluating care. At the time the medical societies were disconcerted by his ideas, which were perceived as revolutionary. After several decade of focusing on the structures and processes of care we are now witnessing a renaissance of measuring outcomes. This paper emphasizes the need for outcomes measurement monitor quality of care. The introduction of diagnosis-related groups in Germany is the most recent development that underlines the importance of outcomes measurement and benchmarking. PMID- 11876051 TI - [Aspects of colchicine therapy. 1: Pharmacology, toxicology, classic indications]. AB - Colchicine has been traditionally used for the treatment of gout. Many observations discuss the prophylactic and/or therapeutic action of colchicine upon a whole range of other diseases. The first part of the overview deals with the pharmacology, and toxicology and the classical indications. PMID- 11876052 TI - [Systemic lupus erythematosus. International Conference on Systemic Lupus Erythematosus, Barcelona, March 24-28, 2001]. PMID- 11876053 TI - [Why a white paper on internal medicine?]. PMID- 11876054 TI - [Pregnancy and internal medicine: a new rubric]. PMID- 11876055 TI - [Diogenes syndrome in the elderly: clinical form of frontal dysfunction? Report of 4 cases]. AB - PURPOSE: Diogenes syndrome is characterised by self-neglect and domestic squalor which leads to unhealthy living conditions. It seems that no single model satisfactorily explains the development of Diogenes syndrome. METHODS: We report four cases of Diogenes syndrome discovered during a short-stay geriatric hospitalization. We were able to analyse them as regards social, physical, psychiatric and cognitive aspects. RESULTS: The results showed absence of socioeconomic precariousness, psychiatric and physical pathologies which generate handicaps. A dementia characterised by a frontal dysfunction was constantly found. CONCLUSION: Diogenes syndrome seems to be a particular behavioural demonstration of a frontal lobe dementia. It does not correspond to a lucid life choice, which must encourage a medico-social assumption. PMID- 11876056 TI - [Changes in hemogram parameters in infections]. AB - PURPOSE: Leukocytosis is considered as an argument for infection. We have compared leukocytosis with the other data available with the white blood cell count. METHODS: White blood cell counts obtained from 187 patients (age: 18 to 81 years), admitted to an emergency room for abdominal pain, were analysed using an automate (Bayer Technicon H2, Dublin, Ireland), and compared with matched healthy subjects. The patients were classified into two groups: diseases of the biliary or of the urinary tract, and subsequently in two subgroups: infectious diseases (angiocholitis, cholecystis, pyelonephritis) or non-infectious diseases (hepatic and nephritic colic). RESULTS: Leukocytes and neutrophils were significantly increased, and eosinophils significantly decreased in all subjects by comparison with controls. These abnormalities were more important in infected patients. Lymphocytes were significantly decreased in infectious disease. For predicting infection, sensitivity and specificity of leukocytosis (> 1,000/mm3) were respectively 66% and 56%, while that of eosinopenia (< 100/mm3) were respectively 91% and 38%, and that of lymphopenia (< 1,200/mm3) respectively 58% and 73%. The probability of infection was less than 3% when neutrophils were less than 7,000/mm3, and eosinophils and lymphocytes respectively more than 100/mm3 and 1,200/mm3. CONCLUSION: This study shows that leukocytosis, eosinopenia or lymphopenia are poor indicators of infection, when considered alone or in combination. However, eosinopenia and lymphopenia appear as better criteria of infection than leukocytosis. A detailed analysis of the white blood cell count allows the exclusion of infection with an acceptable risk of error. PMID- 11876057 TI - [Retrospective study of 55 patients with circulating blood T gama/delta lymphocytosis]. AB - PURPOSE: Gamma/delta T lymphocytes constitute a singular population due to their particular antigenic recognition and their localization inside the epithelium. Their functions are complementary to those of the alpha/beta T-cells and they are involved in the defense and regulation of the immune system. Their role in human diseases is not very well understood and the aim of our study was to analyze a population of patients with a peripheral gamma/delta T-cell lymphocytosis. METHODS: The study included 55 patients, recruited from 1997 to 2000, with a peripheral gamma/delta T lymphocytosis (defined by a proportion of gamma/delta T cells of over 10% of total peripheral T lymphocytes). Analysis of the lymphocyte population was obtained by cytometry after peripheral blood sampling. RESULTS: Three main groups of diseases were observed: infectious diseases (viral infections and tuberculosis), inflammatory diseases (sarcoidosis and autoimmune diseases) and blood diseases (monoclonal gammopathies and hemopathies). Persistence of gamma/delta T lymphocytosis was dependent on the underlying disease (transitional when associated with an infectious disease and lasting when associated with sarcoidosis). The rest of the immunophenotyping analysis was usually normal. CONCLUSION: Our results confirm the data published in the literature concerning the role of the gamma/delta T lymphocytes in infectious, inflammatory and autoimmune diseases and neoplasias. These data are in agreement with the cytotoxic and regular functions of these lymphocytes. PMID- 11876058 TI - [Camptocormia: a sign of axial myopathy. Report of 7 cases]. AB - PURPOSE: Camptocormia or progressive lumbar kyphosis is an anterior bend of the trunk. It appears in orthostatism or while walking and is reducible in the decubitus position. It concerns patients older than 60 years of age. It is due to a fatty degeneration of the paravertebral muscles, although the physiopathology remains unclear. METHODS: We report seven cases of camptocormia revealing authentic myopathies. RESULTS: Our observations concern five women and two men of 55 to 72 years of age. All patients present lumbar kyphosis and had a fatty involution of the paraspinal muscles on the muscular MRI. Four patients fulfilled the Bohan and Peter criteria of polymyositis and dermatomyositis. In the other cases paravertebral muscular biopsies led to the diagnosis of a congenital myopathy, a mitochondrial myopathy and an amyloid myopathy. Four patients received a corticosteroid-immunoglobulins or cyclosporin regimen. An improvement in the camptocormia was observed in three cases. In the other cases the treatment consisted of chemotherapy on account of severe nephrotic syndrome, a coenzyme-Q treatment for the patient with mitochondrial myopathy and only physiotherapy in the case of congenital myopathy, but without positive effect on camptocormia. CONCLUSION: Camptocormia appears as a muscular symptom that may reveal an axial myopathy due to multiple and varied pathologies. Thus, the discovery of camptocormia requires an aetiological investigation in order to propose an adequate treatment, which should be associated with physiotherapy. PMID- 11876059 TI - [Castleman's disease in patients infected with HIV]. AB - PURPOSE: Castleman's disease is a polyclonal lymphoplasmacytic and vascular proliferation prominant in lymphoid tissues. It is associated with lymph node enlargement, hepatosplenomegaly and fever. This manifestations could be secondary to hyperproduction of interleukin 6. The prognosis is poor. The opportunistic infections which are characteristic of severe HIV infection worsen the prognosis. Prolonged monochemotherapy with vinblastine or etoposide can control Castleman's disease. CURRENT KNOWLEDGE AND KEY POINTS: Recent advances in human herpesvirus 8 (HHV8) knowledge and its predominance in the forms which are linked to the HIV seropositivity have partly explained the clinical manifestations of Castleman's disease. Indeed, HHV8 produce an homologous interleukin 6, the vIL-6, responsible for lymphoplasmacytic proliferation. The presence of other homologues of human cytokines produced by HHV8 could contribute to lymphoplasmacytosis and to endothelial proliferation. FUTURE AND PROSPECTS: Taking into account this viral origin, alpha interferon could be an alternative in forms which are less progressive. However, antiviral therapy against HHV8 or HIV and the immunitary restoration do not have any influence on the evolution of Castleman's disease, contrary to opportunistic infections. PMID- 11876060 TI - [Reversibility of radiation-induced fibroatrophy]. AB - PURPOSE: The radiation-induced fibro-atrophic process described in numerous tissues and organs is a localized and irreversible late effect of high-dose radiation therapy. Our purpose is to show that this process is today reversible. CURRENT KNOWLEDGE AND KEY POINTS: This review describes a synthesis of various clinical, paraclinical and histopathological aspects of radiation-induced fibro atrophic process, and of cellular and molecular process regulation. Schematically, there exists a prefibrotic aspecific inflammatory phase, then a constituted and cellular phase, then a matricial densification and remodeling phase, associated in some cases with a tissular terminal necrosis. The respective parts and their evolution during time of the main protagonists as myofibroblast, extracellular matrix and growth factor TGF beta 1 are clarified. From the pathophysiological mechanisms described, curative therapeutic attitudes are proposed for the different progressive phases. Especially, superoxide dismutase (not available) and the pentoxifylline-tocopherol combination seem to allow reduction and reversibility of the fibro-atrophic radiation-induced established process, in clinics as in animal experiments. FUTURE PROSPECTS AND PROJECTS: Some phase II trials try to assess the therapeutic interest of combined pentoxifylline tocopherol in various radiation-induced sequelae, as in osteo-radionecrosis. A clinical randomized trial phase III has just been achieved and could support the results of these experimental and retrospective clinical trials. PMID- 11876061 TI - [Acute alveolo-interstitial pneumopathies in HIV-negative patients treated with antineoplastic chemotherapy]. AB - INTRODUCTION: Treatment of malignant tumors can delay some opportunistic infections. In this paper, we report three cases of alveolo-interstitial pneumonia among HIV negative patients who received chemotherapy. EXEGESIS: All three patients received corticotherapy for a long time. In two cases, it was pneumocystis-tuberculosis coinfection. Evolution was favourable for two patients with sulfamethoxazole-trimethoprime (one with corticotherapy). CONCLUSION: The possible role of chemotherapy, corticotherapy and malignant tumor in the pathogenesis of opportunistic infections will be discussed. PMID- 11876062 TI - [Hereditary persistence of alpha-fetoprotein: report of a case]. AB - INTRODUCTION: Hereditary persistence of alpha-fetoprotein is a rare disorder which exists with no simultaneous disease. The tenth case in the world (two brothers with seminoma and their father) is documented and a complete literature review was done. EXEGESIS: It is transmitted as an autosomal dominant; a disease point mutation has been identified. The failure to recognize hereditary persistence of alpha-fetoprotein sometimes involves unjustified treatments. CONCLUSION: The occurrence of this situation is probably underestimated and it could explain residual levels of alpha-fetoprotein. PMID- 11876063 TI - [Hemolytic-uremic syndrome complicating a long-term treatment with gemcitabine. Report of a case and review of the literature]. AB - INTRODUCTION: Gemcitabine is a nucleoside analog used in solid tumors since 1987. The main side effect is myelosuppression. Acute renal failure with thrombotic microangiopathy has also been reported. We report a new case and suggest to screen for this complication. EXEGESIS: A 71-year-old man with metastatic adenocarcinoma of the pancreas was treated with gemcitabine. He developed episodes of recurred haemolysis followed by haemolytic uremic syndrome. One single haemodialysis session was performed. No other known causes for haemolytic and uremic syndrome were found. Gemcitabine appears to be a new cause of thrombotic microangiopathy. It results from cumulative effects, arises preferentially when there is a renal dysfunction and diagnosis is often delayed. Treatment must be stopped. CONCLUSION: We suggest that reticulocyte count, haptoglobin level and urinalysis could help the clinician to maintain high vigilance and to have a rapid diagnosis for this rare disorder. PMID- 11876064 TI - [Primary orthostatic tremor]. AB - INTRODUCTION: Tremor is frequent in neurologic practice but primary orthostatic tremor was first described in 1984. Its prevalence accounts for around 4% for tremors explored in neurophysiology; in contrast, essential and parkinsonian tremors represent respectively 28 and 22% of these cases. EXEGESIS: Orthostatic tremor is characterized by its appearance while standing. Walking, sitting, and lying down are unaffected. Clinical examination is normal except for unsteadiness disappearing when walking. Surface electromyography in the standing position is necessary for the diagnosis and shows a regular rapid tremor (frequency around 14 to 18 Hz). Its pathophysiology is unknown. CONCLUSION: Clonazepam is the first line treatment for orthostatic tremor. In cases of resistance or side effects of this drug orthostatic tremor may be improved by primidone or, as in our case, gabapentin. PMID- 11876065 TI - [Unusual association: hepatic peliosis and Crohn's disease]. AB - INTRODUCTION: Hepatobiliary disorders occurs in about 10 to 30% of inflammatory bowel diseases. Fatty liver and gallstones are preferentially described with Crohn's disease. Conversely, peliosis hepatis has been only exceptionally reported in association with inflammatory bowel diseases. We describe here a case of peliosis hepatis complicating a misdiagnosed Crohn's disease. EXEGESIS: A 24 year-old woman was referred because of a hepatomegaly and a deterioration of the general state. The patient complained about diarrhea for 3 months and about an episode of bloody stools. There was an aphthous-like lesion on the tongue. X-ray pull-through revealed a typical aspect of terminal ileitis as seen in Crohn's disease. The liver was heterogeneous. Liver biopsy revealed a peliosis hepatis. No other possible cause of peliosis hepatis was found. Therefore, in this case, peliosis hepatis, revealed by the hepatomegaly, was associated with Crohn's disease. CONCLUSION: Association between Crohn's disease and peliosis hepatis has only exceptionally been described. A priori, this complication is not a pejorative feature in inflammatory bowel disease. The pathophysiological significance of this association remains unknown. PMID- 11876066 TI - [Recurrent cerebral hematoma]. PMID- 11876067 TI - [Lyme disease disclosed by pseudoneoplastic weight loss]. PMID- 11876068 TI - [Breast plasmacytoma. A new case]. PMID- 11876069 TI - [Serous non-Hodgkin's lymphoma in immunocompetent patient]. PMID- 11876070 TI - Multivariate ordination approach for identification of sub-regional homogeneities in Gujarat, western India. AB - Identification of homogeneous invariant geographical units assumes considerable importance in the context of current attempts at developing environmental indices to monitor changes. Data on the edaphic and climatic parameters for the state of Gujarat--a predominantly arid to semi-arid province in western India--has been used to elaborate an approach that employs principal component analysis (PCA) as the basis for unbiased clustering of similar geographic units at sub-regional scales. PCA identified different sets of edaphic and climatic variables responsible for separating one unit from another. With the exception of a few cases, soil variables exhibited greater influence on the separation of units than climate. Response of some agro-ecological variables (total cultivable area, forestland and normalized difference vegetation index) to both original and ordinated variables in each of the identified edaphic-climatic units (ECU) has been evaluated using step-wise multiple regressions. The edaphic-climatic parameters were good predictors of these variables, although ordination did not necessarily enhance the predictive power of the independent variables. Instead, in many cases original variables showed better correlations. The response of these variables, chosen on the basis of varying levels of dynamism (slow to seasonal fluctuations), showed that independent of the degree of dynamism, the dependence of these variables was distinct in each ECU. This provided a reality check on the agro-ecological and bio-climatic differentiation among the ECUs. PMID- 11876071 TI - Using Geographical Information Systems to identify and target sites for creation and restoration of native woodlands: a case study of the Chiltern Hills, UK. AB - Rare and threatened habitats in Europe must be restored and enhanced in accordance with the European Union's Habitats and Species Directive. In the United Kingdom, conservation and expansion objectives for species and habitats are outlined in the Species Action Plans and Habitat Action Plans. Site identification for these measures has to date been ad hoc without consideration of either the existing "stock" of the natural resource or the ability of the surrounding land use to deliver the enhancement (enlargement) of a given habitat. Using a Geographical Information System, we outline a targeting system for creating new woodland in association with existing ancient woodland in the Chiltern Hills Area of Outstanding Natural Beauty. The aim was to create woodland blocks of at least 100 ha, as being of the most benefit to biodiversity. We identified existing patches of woodland between 20 and 50 ha as cores for habitat expansion and classified land use in terms of its suitability and proximity to these core areas for tree planting to meet the targets of the statutory body. Our results suggest that the targeting method employed is a useful tool for habitat restoration. PMID- 11876072 TI - More woody plants? The status of bush encroachment in Botswana's grazing areas. AB - Foci points, which are currently intensified by increased anthropogenic activities, have resulted in vegetation changes in the cattle-dominated grazing areas of Botswana. Bush encroachment species--for instance Acacia tortilis, A. erubescens, A. mellifera, Dichrostachys cinerea, Grewia flava, and Terminalia sericea--are increasing in cover and density around foci points (e.g. water points and kraals) at the expense of the grass cover. A number of factors have the effect of encouraging the germination and survival of bush encroachment species. The practice of cattle husbandry and continual shifting of foci points within grazing areas have resulted in the spread of the distribution of bush encroachment species across the country. This is evidenced by the potential extent of 37,000 km2 (6.4% of Botswana) of darkened and near infrared (NIR) reflective bush encroached areas in 1994. This paper suggests that specific management strategies should be adopted to help overcome the bush encroachment problem, which is causing a significant reduction in the extent of Botswana's high quality rangeland. These strategies may vary from the enforced reduction of grazing intensity in areas identified as being heavily bush encroached to the selective management of opportunistic (communal) grazing in better quality predominantly grassland areas. Further work is however required to update this analysis and especially to consider trends since 1994-1995. While some work on the extent of woody cover and the further causes of bush encroachment is being undertaken under the SAFARI2000 project, more research is needed in specific areas to pinpoint causes and responses to the bush encroachment problem. PMID- 11876073 TI - Preferences for dealing with environmental problems: an empirical study of managers in three mainland Chinese cities. AB - This study examines managerial preferences for various policy mechanisms commonly used to address environmental problems using a sample of 653 Chinese managers from firms in Beijing, Dalian and Guangzhou. Overall, these managers appeared to favor regulatory instruments, placing somewhat less faith in market and communicative instruments. However, all measures appear to have ample support within each subgroup. In addition, the influence of two situational factors on these preferences was investigated--the immediacy of environmental impact and diffusion of responsibility. The results of the study indicate that these two factors interact in explaining managerial preferences, but only for the use of market mechanisms. PMID- 11876074 TI - Development and implementation of the Norwegian monitoring programme for agricultural landscapes. AB - This paper describes the development and implementation of the Norwegian monitoring programme for agricultural landscapes--the '3Q programme'. The main objective of the scheme is to indicate development trends in the agricultural landscape, and their consequences for spatial structure, biodiversity, cultural heritage and accessibility. The monitoring programme aims to give policy feedback and provide data to fulfill international reporting requirements. This paper describes the background to the programme and reasons behind the choice of methods. Results are presented to show the accuracy of the methods employed and the range of indicator values recorded in the programme. Strengths and limitations of the monitoring programme are discussed, and potential future improvements and developments are outlined. Although there remains a potential for methodological improvement, we stress the importance of establishing a baseline to enable the detection of development trends in a rapidly changing environment. PMID- 11876075 TI - Urbanization's impact on timber harvesting in the south central United States. AB - The impact of urbanization on timber harvesting in the south central United States was investigated. Geo-referenced Census and Forest Inventory Analysis (FIA) data were combined using a geographic information system (GIS) in order to examine the effects of various demographic and biophysical forest inventory characteristics on timber harvesting. These effects were estimated for intermediate and final harvests using a multinomial logit model. The probability for both types of harvests decreased with increasing population density, decreasing forest size, and decreasing distance to urban areas; however, the reduction in intermediate harvests was greater for each variable. Harvesting rates decreased by as much as 19% as population densities increased or distance to urban areas decreased. The results indicated that active forest management is curtailed far beyond the urban boundary. In order to model the impact of urbanization adequately, timber supply projections must also account for its impact on harvesting frequencies in surrounding areas. PMID- 11876076 TI - Cross-achievements between policies for drinking water protection. AB - Environmental dynamics have important spatial dimensions, which calls for a spatial approach in policy analyses. Further to this, assessing agri environmental policies involves analyses of individual measures as well as their combined effects on farmer behaviour and the environment. The integration of an economic behavioural model in a spatial framework has enabled analyses of a geographically targeted subsidy scheme for drinking water protection in combination with a uniform tax on commercial nitrogen fertilizer. Results show that policy measures for reducing nitrogen use can have combined effects (cross achievements), thereby affecting each other's cost-effectiveness. Cross achievements between a nitrogen fertilizer tax and a subsidy scheme based on elicitation are shown not to be additive, making partial analyses of policy measures more uncertain. PMID- 11876077 TI - The application of local measures of spatial autocorrelation for describing pattern in north Australian landscapes. AB - This paper tests the use of a spatial analysis technique, based on the calculation of local spatial autocorrelation, as a possible approach for modelling and quantifying structure in northern Australian savanna landscapes. Unlike many landscapes in the world, northern Australian savanna landscapes appear on the surface to be intact. They have not experienced the same large scale land clearance and intensive land management as other landscapes across Australia. Despite this, natural resource managers are beginning to notice that processes are breaking down and declines in species are becoming more evident. With future declines of species looking more imminent it is particularly important that models are available that can help to assess landscape health, and quantify any structural change that takes place. GIS and landscape ecology provide a useful way of describing landscapes both spatially and temporally and have proved to be particularly useful for understanding vegetation structure or pattern in landscapes across the world. There are many measures that examine spatial structure in the landscape and most of these are now available in a GIS environment (e.g. FRAGSTATS* ARC, r.le, and Patch Analyst). All these methods depend on a landscape described in terms of patches, corridors and matrix. However, since landscapes in northern Australia appear to be relatively intact they tend to exist as surfaces of continuous variation rather than in clearly defined homogeneous units. As a result they cannot be easily described using entity-based models requiring patches and other essentially cartographic approaches. This means that more appropriate methods need to be developed and explored. The approach examined in this paper enables clustering and local pattern in the data to be identified and forms a generic method for conceptualising the landscape structure where patches are not obvious and where boundaries between landscape features are difficult to determine. Two sites are examined using this approach. They have been exposed to different degrees of disturbance by fire and grazing. The results show that savanna landscapes are very complex and that even where there is a high degree of disturbance the landscape is still relatively heterogeneous. This means that treating savanna landscapes as being made up of homogeneous units can limit analysis of pattern, as it can over simplify the structure present, and that methods such as the autocorrelation approach are useful tools for quantifying the variable nature of these landscapes. PMID- 11876078 TI - [The quality of life of chronically ill and psychiatrically disturbed children. Initial experiences with an inventory for assessing quality of life in children and adolescents]. AB - OBJECTIVES: The assessment of the quality of life in chronically ill children as well as in children with emotional and behavioural problems. METHODS: A series of questionnaires (Inventar zur Erfassung der Lebensqualitat bei Kindern und Jugendlichen, ILK, Mattejat et al.) were administered to 360 patients and 288 mothers at the Vienna University Children's Hospital. RESULTS: The results clearly show that patients with diabetes rate themselves as the most emotionally burdened and impaired in nearly all aspects of life: a picture which is confirmed by their mothers. Children and adolescents with psychological problems judge the initial problem as well as their social environment as significantly more burdensome. The respective mothers of these psychologically ill children feel that their impairments are greater in more aspects of life than do mothers of children who are chronically ill. The children generally pinpoint most of their difficulties in the areas "ability to occupy myself" and "psychological health". Among adolescents the most negative ratings occur in the areas "school", "psychological health", and "initial problem". CONCLUSIONS: The child's problems seem to weigh heavily upon the mothers. Interventions aimed at adolescents and mothers should be based on problem solving: supportive and anxiety-reducing approaches seem best suited for chronically ill children. Children with psychological problems primarily need problem-centred and practical support. PMID- 11876079 TI - [Specificity hypothesis of a theory of mind deficit in early childhood autism]. AB - OBJECTIVES: In order to test the hypothesis that a theory of mind deficit is specific for autism, the present study presents the first replication of the Sally-Anne test (Baron-Cohen, Leslie & Frith, 1985) in the German-speaking countries. METHODS: The Sally-Anne test was administered to 16 autistic, 24 probands with Down's syndrome and 20 normal preschool prosands. The intelligence of the autistic group and that with Down's syndrome was measured by the CPM/SPM. In addition, the ADI-R was used with the principal caregivers of the autistic and Down's syndrome subjects. RESULTS: With regard to the clinical diagnosis, theory of mind deficit turned out to be not specific for autism. Six of 16 (37.5%) autistic subjects passed the theory of mind tasks. Thus performance in the autistic group surpassed that of both control groups. Out of 16 autistic subjects, autism could be confirmed in only 8 on the basis of the ADI-R diagnostic criteria, only one of whom showed a theory of mind. The autistic individuals with a theory of mind differed significantly in their mean IQ from those without this ability. CONCLUSIONS: Spectrum and specificity of a theory of mind deficit in autism remain controversial. For further research it seems important to administer the ADI-R during the diagnostic process. The findings suggest that the clinical diagnosis of autism is not precise enough to distinguish between autism and nonautistic mental handicap. PMID- 11876080 TI - [The Baer law in the age of biotechnology]. PMID- 11876081 TI - [Behavior therapy in children with aggressive dyssocial disorders]. AB - Conduct disorder in children and adolescents is prevalent and has a poor prognosis: it is now considered to be a chronic condition that requires treatment as well as life-long monitoring. Two forms of behavior therapy, parent management training and training in problem-solving skills are currently seen as the most effective treatment. Parent management training is the treatment of choice for younger children, while for older children it should be supplemented with training in problem-solving skills. Parent management training is one of the best researched therapy techniques for conduct disorder, deserving the criterion "empirically supported treatment" defined by the American Psychological Association. Training in problem-solving skills has led to therapeutic change in clinical samples and is qualified as "probably efficacious treatment". Limitations of both treatments include the high drop-out rate among parents and the fact that many youth improve but remain outside the range of normative functioning. To optimize treatment effectiveness, complex prevention programs have been developed in recent years. Results to date are mixed; however, two large ongoing studies, the FAST and the LIFT study, report positive first outcomes. A higher priority of conduct disorder and related problems in child psychiatric and psychotherapeutic services would be desirable. PMID- 11876082 TI - [Motor, cognitive and socio-emotional development of 11-year-olds with early childhood risk factors: late sequelae]. AB - OBJECTIVES: The investigation of the impact of early childhood risk factors on developmental outcome at the age of 11 years. METHODS: The onset and course of developmental and behavioral disturbances were examined in a prospective longitudinal study of a sample of 362 children born with different risks. Organic (obstetric complications) and psychosocial risks (family adversity) were varied in a two-factorial design. Measures of motor, cognitive and social-emotional outcome were obtained from 341 children aged 11 years (168 boys, 173 girls, corresponding to 94.2% of the initial sample). Previous assessments had been conducted at the ages of 3 months, and again at the ages of 2, 4 1/2 and 8 years. RESULTS: The negative impact of early risk factors persisted into late childhood. Rates of developmental and behavioral disturbances in high-risk children were up to three times higher than in non-risk children. Both organic and psychosocial risks contributed to adverse outcomes. While organic complications were related to disturbances in motor and cognitive development, the detrimental effects of psychosocial adversity pertained to social-emotional functioning. Late sequelae of pre- and perinatal complications were found especially in cognitive outcome and school performance. The cumulative effect of early risks was best explained by summing up the single risk effects. CONCLUSIONS: Early risk factors have specific and long-term sequelae resulting in adverse school outcomes at later ages. PMID- 11876083 TI - [Is topiramate effective for weight loss in neuroleptic-induced obesity? 2 case reports]. AB - OBJECTIVES: Topiramate, a fairly new anticonvulsant, is increasingly being used as a mood stabilizer in bipolar and schizoaffective disorders. One common side effect is a reduced appetite that often results in weight loss. This finding raises the interesting question of whether both mood and body weight can be stabilized in patients who have gained weight while being treated with neuroleptics for one of the disorders mentioned above. METHODS: We studied the body weight, subjective sense of well-being, and psychopathology in two adolescent patients who were being treated with topiramate (alone or in combination with a neuroleptic drug). Both patients had reduced appetite, while body weight either remained stable or was reduced. RESULTS: The patients reported both improved control of food intake and mood stabilization. CONCLUSIONS: We conclude that adolescents with affective disturbance who have gained weight on neuroleptic drugs may benefit from topiramate in terms of mood stabilization and body weight control. PMID- 11876084 TI - [Early childhood psychosis versus early childhood autism? Differential diagnosis based on one week observation in an inpatient setting]. AB - We report on the case of a 4-year old child suffering for the past six months from a lingering psychotic development. Several months of outpatient observation did not permit a clear differential diagnosis of either childhood autism or an early psychosis. During a one-week inpatient stay at our hospital recurring productive-psychotic episodes were observed that finally led to the exclusion of the diagnosis of "childhood autism" (ICD-10 F 84.0 (Dilling et al., 1993)) and to the diagnosis of "other disintegrative childhood disorder" (ICD-10: F 84.3 (Dilling et al., 1993)). PMID- 11876086 TI - Healthy Delaware 2010. PMID- 11876087 TI - Perspective on preventive medicine in the 21st century. PMID- 11876085 TI - Collagen-binding I domain integrins--what do they do? AB - Collagens are the most abundant proteins in the mammalian body and it is well recognized that collagens fulfill an important structural role in the extracellular matrix in a number of tissues. Inactivation of the collagen alpha 1(I) gene in mice results in embryonic lethality and collagen mutations in humans cause defects leading to disease. Integrins constitute a major group of receptors for extracellular matrix components, including collagens. Currently four collagen binding I domain-containing integrins are known, namely alpha 1 beta 1, alpha 2 beta 1, alpha 10 beta 1 and alpha 11 beta 1. Unlike the undisputed role of collagens as structural elements, the biological importance of integrin mediated cell-collagen interactions is far from clear. This is in part due to the limited information available on the most recent additions of the integrin family, alpha 10 beta 1 and alpha 11 beta 1. Future studies using gene inactivation of individual and multiple integrin genes will allow testing of the hypothesis that collagen-binding integrins have redundant functions but will also shed light on their importance in pathological conditions. In this review we will describe what is currently known about the collagen-binding integrins and discuss their biological functions. PMID- 11876088 TI - Medicine and public health. PMID- 11876089 TI - Childhood obesity: evaluation, treatment, and relationship to disease. PMID- 11876090 TI - Evidence-based approach to preventive care. PMID- 11876091 TI - The case for screening for colorectal cancer: a preventable disease. PMID- 11876092 TI - Preventing health problems: in the office and legislative hall. PMID- 11876093 TI - Births: final data for 2000. AB - OBJECTIVES: This report presents 2000 data on U.S. births according to a wide variety of characteristics. Data are presented for maternal demographic characteristics including age, live-birth order, race, Hispanic origin, marital status, and educational attainment; maternal characteristics (medical risk factors, weight gain, tobacco and alcohol use); medical care utilization by pregnant women (prenatal care, obstetric procedures, complications of labor and/or delivery, attendant at birth, and method of delivery); and infant characteristics (period of gestation, birthweight, Apgar score, abnormal conditions, congenital anomalies, and multiple births). Also presented are birth and fertility rates by age, live-birth order, race, Hispanic origin, and marital status. Selected data by mother's State of residence are shown, as well as data on month and day of birth, sex ratio, and age of father. Trends in fertility patterns and maternal and infant characteristics are described and interpreted. METHODS: Descriptive tabulations of data reported on the birth certificates of the 4.059 million births that occurred in 2000 are presented. RESULTS: The number of births rose 3 percent in 2000; birth and fertility rates rose 1 to 2 percent. The total fertility rate was above "replacement" for the first time in almost 30 years. Teenage birth rates continued to fall while birth rates for women aged 20 24 years rose slightly, and rates for women in their late twenties and thirties rose 3 to 5 percent. Births to women in their forties and early fifties were also up for 2000. The number of births to unmarried women, the birth rate, and the percent of births that were to unmarried women rose 1 to 3 percent, but birth rates for unmarried teenagers declined. Smoking by pregnant women was down again. The cesarean delivery rate rose 4 percent to 22.9, the fourth consecutive increase; the primary cesarean rate was up and the rate of vaginal births after a previous cesarean was down. The number and rate of twin births continued to rise, but the triplet/+ birth rate declined for the second year in a row. For the first year in almost a decade the preterm birth rate declined (to 11.6 percent); the low birthweight rate, however, was unchanged at 7.6 percent. PMID- 11876094 TI - The history of immunization in the U.S. armed forces. PMID- 11876096 TI - A good shot. PMID- 11876095 TI - New directions in vaccine research. PMID- 11876097 TI - No shots for my child! PMID- 11876098 TI - Childhood vaccines--what's new? PMID- 11876099 TI - Vaccinating adults for vaccine-preventable diseases. PMID- 11876100 TI - Vaccines in public health emergencies. PMID- 11876101 TI - Not my fault--call a lawyer. PMID- 11876102 TI - Giving vaccines a boost. PMID- 11876103 TI - Improving Web site ease-of-use for people with disabilities. PMID- 11876104 TI - Marketing tip: using usenet. PMID- 11876106 TI - Search engine strategies. PMID- 11876105 TI - Can quality seals save us from the sharks? PMID- 11876107 TI - Improving usability: site maps. PMID- 11876108 TI - California hospital system launches Web-based asthma program. PMID- 11876109 TI - Fetal thoracic measurements in prenatal diagnosis of Jeune syndrome. AB - We describe prenatal sonographic findings in a 34-week fetus with Jeune syndrome or asphyxiating thoracic dystrophy (ATD). The long bones measured were less than third percentile; the thoracic circumference (TC) measured 216 mm (< 2.5th percentile); the abdominal circumference (AC) measured 303.5 mm (50th-75th percentiles) and the rib cage perimeter (RCP) measured was 98 mm. The TC/AC was 0.70 (normal, 0.85) and the RCP/TC was 0.45 (normal, 0.68). Following birth diagnosis of Jeune syndrome was made based on radiographic analysis, which was subsequently confirmed by clinical and postmortem examination. This case highlights the utility of both TC/AC and RCP/TC in diagnosis of ATD and other skeletal dysplasias associated with a small thorax. PMID- 11876110 TI - Neonatal Bartter syndrome. AB - A case of neonatal Bartter syndrome is reported. The baby born pre-term following a pregnancy complicated by polyhydramnios, presented at 7 months of age with failure to thrive, gastroenteritis and facial dysmorphisms. An unusual feature was the absence of the classical biochemical abnormality of hypochloremic alkalosis early in the course of the disease. Metabolic acidosis was the initial manifestation at 5 weeks of age. Awareness of this presentation is important to avoid delay in diagnosis and treatment. PMID- 11876112 TI - Supra-patellar bursitis in a neonate. PMID- 11876111 TI - Morquio-B syndrome (MPS-IV B) associated with beta-galactosidase deficiency in two siblings. AB - In the present article we describe two cases with Morquio-B syndrome characterized by beta-galactosidase deficiency in a Muslim family. They were found to have skeletal dysplasia, short stature and short trunk dwarfism with undetectable level of beta-galactosidase in leucocytes. Probands' sister who had no clinical signs of mucopolysaccharidosis was investigated and found to have normal levels of the enzyme. Mother was found to have a deficient activity of beta-galactosidase and father was not available for the study. Since mother was pregnant, prenatal study from chorionic cells was carried out to investigate beta galactosidase activity in the chorionic villus. An intermediate level of beta galactosidase activity was found in the chorionic villus cells suggesting a carrier status. The diversity and rarity of the study makes it worth presenting. PMID- 11876113 TI - Conventional ventilation in neonates: experience from Saudi Arabia. AB - OBJECTIVE: To study the clinical profile and immediate outcome of inborn neonates receiving intermittent positive pressure ventilation (IPPV) at the neonatal intensive care unit of Civil Hospital, Khamis Mushayt, Saudi Arabia, a level II nursery. METHODS: 78 liveborn neonates who had received IPPV over a 20 months period from January 1999 to August 2000 were reviewed from their charts and nursery registers. The indications for IPPV and the immediate outcome including complications were studied with respect to various weight groups (1 kg or less, > 1-1.25 kg, > 1.25-1.5, > 1.5-2 kg and > 2 kg) and gestation groups (28 weeks or less, 29-32 weeks, 33-36 weeks and full term). RESULT: Hyaline Membrane disease (n = 31, 39.7%) and perinatal asphyxia (n = 29, 37.2%) were the major indications for IPPV. 67.9% (53 of the 78) ventilated neonates survived. The chances for survival showed a statistically significant increase with increasing birthweight (P = 0.0006) and with increasing gestational age (P = 0.002). (80%) (44 of 55) of neonates weighing more than 1.25 kg survived vs 39.1% (9 of 23) of those 1.25 kg or less, P = 0.0011. Similarly, 79.3% (46 of 58) of neonates of 29 or more weeks of gestation survived vs 35% (7 of 20) of those 28 weeks or less, P = 0.0007. The complications seen in the study group included blood culture positive sepsis (n = 7), pulmonary hemorrhage (n = 6), air leak syndromes (n = 4), endotracheal tube related problems (n = 5), chronic lung disease (n = 3) and retinopathy of prematurity (n = 2). CONCLUSION: Gestational age of less than 28 weeks and birth weight less than 1.25 kg can be recommended as the cut off weight and gestation criteria for in utero transfer in this centre and upgradation of existing facilities are urgently called for to improve the survival rates further. PMID- 11876114 TI - Determinants of pregnancy in adolescents in Nepal. AB - OBJECTIVE: Social factors and prevalent norms in the community determine the proportion of teenage pregnancy in the community. In the light of high risk associated with teenage pregnancy, the socio-cultural determinants, which influence the conception among teenagers in Nepal, need to be understood. These determinants may be modified by suitable interventions to reduce teenage pregnancy. Aim of this study was to examine the socio-cultural determinants of teenage pregnancy in eastern Nepal. METHODS: A case-control study design was selected for comparing the education, economic status, family support and freedom towards conception among teenagers as compared to higher age group women. RESULT: Seventy adolescent pregnant women were compared with seventy primigravida women in the 20 to 29 years age group. The teenage pregnant women were less educated, had poor economic background, more likely to have accidental pregnancies as compared to the other group and more likely to have love marriages. Husbands were more likely to decide about continuation of pregnancy. They had less psychological and social support from the family. CONCLUSION: Marriage at young age and pregnancy during teens are associated with less social acceptance and poor support in the family. PMID- 11876115 TI - Autopsy study of pediatric deaths. AB - OBJECTIVE: Childhood autopsies occurring during January 1997 to December 1999 were reviewed. METHOD: Of the total 1,445 deaths autopsied, 441 (30.5%) were of pediatric patients. RESULT: Of these deaths, 70.3% occurred during neonatal period, 8.6% in infancy, and 21.1% in childhood. Birth asphyxia (57.7%) was the most frequent mode of death in the neonatal period. Infections (67.2%) predominantly accounted for infant and childhood mortality, those of respiratory tract (46.1%) being most frequent in infancy, early and mid childhood, and enteric fever in late childhood (33.3%). Discrepancy between ante-mortem and post mortem diagnosis occurred in 29 cases (6.6%). CONCLUSION: Autopsy is an important clinical tool providing useful information to the physician. Few published reports are available on pediatric autopsies. This study aims to help in a better understanding of causes of deaths in neonates, infants and children. PMID- 11876116 TI - Quantification of coronary risk score. AB - OBJECTIVE: To quantify the coronary risk score in children and/or grandchildren of patients with coronary artery disease (CAD). METHODS: One hundred and fifty children with positive family history of CAD (cases) and age and sex matched children with no such history (controls) were enrolled in the study. Fasting blood sugar, serum cholesterol and triglycerides were estimated on all children and a risk score was calculated using Nora's scheme. RESULT: The cases and controls were similar with respect to mean age, six distribution, mean weight and mean height. None of the child was hypertensive. Mean serum levels of triglycerides were 164.7 m/dl and 105.7 mg% respectively in cases and controls (p < 0.005). Prevalence of hypertriglyceridemia was three times more in cases (p < 0.001). Cases had a significantly higher risk scores (p < 0.00001) and this was mainly attributable to higher biochemical risk (P < 0.0006). CONCLUSION: Children and grand children with a positive family history of CAD had significantly higher risk scores for future CAD. The increased risk was mainly due to biochemical derangements. PMID- 11876117 TI - Bone marrow transplantation for hematological disorders--Shiraz experience. AB - OBJECTIVE: In the past 8 years, 120 cases of hematological disorders were transplanted from the HLA identical donors. METHOD: Using chemotherapy based conditioning regimen with cyclophosphamide 200 mg/kg and busulfan 15-16 mg/kg, 80 cases of beta-thalassemia major and 35 cases of leukemia and five patients with aplastic anemia had received bone marrow transplantation. RESULT: The five-year survival in thalassemic group was 72%, for leukemic group (acute and chronic) was 58%, and also for aplastic anemia 65%. Transplantation related mortality was the cause of death in 29 cases. The two major causes of death were acute graft versus host disease and poor medical condition of patients before marrow transplantation. CONCLUSION: At the present time, allogenic marrow transplantation is curative mode of treatment for many hematological diseases. PMID- 11876118 TI - Measles outbreak in a Periurban area of Chandigarh: need for improving vaccine coverage and strengthening surveillance. AB - OBJECTIVE: An outbreak of measles was investigated in the periurban areas of Chandigarh Union Territory, during the months of December 1998 to February 1999. Mainly the children below 15 years of age were affected. The children of migrant labourers belonging to the neighbouring states of Uttar Pradesh and Bihar constituted the majority of population in the area under study. They belonged to lower socio economic status with low immunization coverage. METHODS: A total of 2968 houses were surveyed for epidemiological investigations in the areas of colony No. 5, Ramdarbar, Palsora and Pandit colony of Kajheri, covering a population of 14,601 and 7.3% (216/2968) of families were affected in the outbreak. RESULTS: Two hundred and eighty three cases of measles were reported with an attack rate of 4.5% and male to female ratio of (M:F) 5.3%:3.6%. Among the measles cases, 48.8% had received measles vaccination. The outbreak was investigated by detecting measles specific IgG/IgM antibodies either in acute or convalescent serum samples or both. Due to inadequate surveillance system and containment measures, the outbreak was in full swing during the winter months. Measles related complications were reported in 31.1% cases (i.e. diarrhoea in 15.2% and Pneumonia is 7.1%). CONCLUSION: Following smallpox and guinea worm eradication, WHO's next thrust, is on eradication of poliomyelitis and measles. Hence, strengthening of disease surveillance as well as vaccination policies are mandatory to achieve disease control in these areas. PMID- 11876119 TI - Do we need campaign approach of vitamin A administration in non vitamin A deficient areas? PMID- 11876120 TI - Integrated Management of Childhood Illness (IMCI): a robust strategy. AB - Integrated Management of Childhood Illness (IMCI), a strategy fostering holistic approach to child health and development, is built upon successful experiences gained from effective child health interventions like immunization, oral rehydration therapy, management of acute respiratory infections and improved infant feeding. The core intervention of IMCI is integrated management of the five most important causes of childhood deaths-acute respiratory infections, diarrheal diseases, measles, malaria and malnutrition. Using a set of interventions for the integrated treatment and prevention of major childhood illnesses, the IMCI strategy aims to reduce death as well as the frequency and severity of illness and disability, thus contributing to improved growth and development. In health facilities, the IMCI strategy promotes the accurate identification of childhood illness (es) in the outpatient settings, ensures appropriate combined treatment of all major illnesses, strengthens the counselling of caretakers and the provision of preventive services, and speeds up the referral of severely ill children. The strategy also aims to improve the quality of care of sick children at the referral level. It also creates a scientifically sound link between the management guidelines at the community level and the management approach in a referral centre. The strategy also envisages actual situations when referral is not possible and offers the best possible options in such circumstances. In the home setting, it promotes appropriate early home care and care-seeking, improved nutrition and prevention, and the correct implementation of prescribed care. In addition to its focus on treatment of illness in the health facility as well as at home, it also provides an opportunity for important preventive interventions such as immunization and improved infant and child nutrition including breastfeeding. The IMCI strategy reduces wastage of resources and avoids duplication of efforts that may occur in a series of separate disease control programs. The essential pillars include improvement in the case management skills of health personnel, improvement in health systems, and improvement in family and community practices. IMCI has been introduced in more than 80 countries and 19 of them have already scaled up IMCI implementation Even though it is too early to relate the decrease in childhood mortality with the introduction of IMCI in these countries, there are several indirect indicators which endorse its validity as a comprehensive and effective strategy. IMCI has helped countries to revise and update their child health policies, streamline the essential drug lists for children, increase service utilization, improve quality of care and nutritional counselling, improve health systems and improved family and community practices. PMID- 11876121 TI - Anthrax. AB - Anthrax is caused by Bacillus anthracis, an encapulated and spore-forming bacillus. The disease is usually contracted through uptake of spores that remain viable in the contaminated soil for many years. Anthrax is primarily a disease of herbivorous animals and is uncommon in humans who may get the infection through contact with contaminated animals or their products. Anthrax spores germinate after entering the body through skin abrasions (cutaneous anthrax) or by inhalation (inhalation anthrax) or ingestion (gastrointestinal anthrax) and multiply to produce two exotoxins which determine the virulence along with capsule. Although most cases occur within 48 hours of exposure, germination of spores may occur upto 60 days later. While inhalation anthrax is almost always fatal, intestinal anthrax results in death in 25% to 60% of cases. Upto 20% of cases having cutaneous anthrax may die. Antibiotics are effective if the disease is recognised early and treated appropriately. Penicillin is the drug of choice when disease occurs in natural setting. Ciprofloxacin is recommended when aerosols of anthrax spores are used as bioweapon, prophylactic antibiotics should not be prescribed until risk of exposure is considered real by experts. PMID- 11876122 TI - Atopy Patch Test (APT)--its role in diagnosis of food allergy in atopic dermatitis. AB - During the past few years, the Atopy Patch Test (APT) has become a valuable additional tool in the diagnostic work-up of food allergy in infants and children with atopic dermatitis. A positive APT may help to prevent restrictive and unnecessary diets which may be the consequence of misjudging late reactions by clinical assessment alone. Furthermore, the combination of positive atopy patch tests together with defined levels of specific IgE (cow milk = 0.35 kU/l and egg = 17.5 kU/l) makes DBPCFC superfluous for suspected milk and egg allergy. However, several open questions indicate the need for further studies. PMID- 11876123 TI - Food allergy in children. AB - Food allergies are increasing in prevalence and as a disease burden throughout the world, however they seem to increasingly affect countries with a formerly low prevalence. Consideration and diagnosis of food allergies are important as it has ramifications that affect a child's diet, care at school and home and is recognised to be associated with anxiety of parents, family and care takers. Food allergies vary significantly between countries, however nut allergies appear to be widely associated with serious reactions and death. The value of specific food immunoglobulin E (IgE) and skin prick tests (SPT) has been extensively analysed in children in recent years and can provide very useful information in an appropriately selected population. Diagnosis may require formal challenges to confirm a genuine allergic reaction condition rather than an intolerance reaction due to other mechanisms. The medical care of a food allergic child requires concurrent dietary advice and management, risk avoidance and emergency management plans. Reintroduction of foods occurs when the risk profile is appropriate based mainly on the predictive information obtained by SPTs and specific IgE levels. Fortunately allergies to egg and milk resolve by 3-5 years in about 80% of children. There have been some recent advances in immunotherapeutic approaches to food allergy although this has not translated to success in human treatment to date. PMID- 11876124 TI - Antipyretics in children. AB - Fever is an important symptom of underlying disease condition and in general is considered harmful in pediatric age group as it may lead to febrile seizures, stupor, dehydration increase work of breathing, discomfort and tachycardia. The increase metabolic demands stress the patient with marginal cardiac and cerebral vascular supply. The hypothalamus controls the body temperature. The fever results due to resetting of the hypothalamus that occur from the prostaglandins produced by the pyrogens. Fever is treated variedly by the pediatricians. The physical therapy offers a simple and cost effective way of lowering the body temperature. The drugs as paracetamol, nimesulide and ibuprofen lower the temperature by inhibiting the prostaglandin synthesis. Paracetamol is considered the safest of all the antipyretic drugs. It is recommended that a combination of physical therapy such as tepid sponging and paracetamol is best way of controlling temperature. PMID- 11876125 TI - Diabetic ketoacidosis in the pediatric patient. AB - Diabetic ketoacidosis is the major source of morbidity and mortality in patients with Type I diabetes mellitus. One population-based study noted that approximately 25% of newly diagnosed patients present with diabetic ketoacidosis. Most of those hospitalized with diabetes ketoacidosis, however, are those with poor control of their pre-existing diabetes. Cerebral edema has been found to be present in only 1% of children with diabetic ketoacidosis, but accounts for 20% of all diabetes-related deaths. Although the mortality rate has fallen dramatically over the course of the last century, the underlying mechanisms involved with the development of cerebral edema are still not clear. Several studies have shown specific risk factors to be associated with cerebral edema, but as of yet no reduction in the recent incidence of cerebral edema has been demonstrated. A discussion of these risk factors and a suggested protocol for treating diabetic ketoacidosis are included in this review. PMID- 11876126 TI - Polycythemia in the newborn. AB - Polycythemia is defined as a venous hematocrit above 65%. The relationship between viscosity and hematocrit is almost linear till 65% and exponential thereafter. Increased viscosity of blood is associated with symptoms of hypo perfusion. The hematocrit in a newborn peaks at 2 hours of age and decreases gradually after that. The etiology of polycythemia is related either to intra uterine hypoxia or secondary to fetal transfusion. Clinical features related to hyperviscosity may affect all organ systems and this entity should be screened for in high-risk infants. Polycythemia maybe symptomatic or asymptomatic and guidelines for management of both types are provided in the protocol. PMID- 11876127 TI - Retinopathy of prematurity. AB - With improving survival of very low birth infants in India, Retinopathy of Prematurity (ROP is likely to emerge as a significant problem. The most important risk factor in the pathogenesis of ROP is prematurity. Other factors like frequent blood transfusions; sepsis, apnea and problems with oxygenation have also been implicated in the causation of ROP. Essentially asymptomatic in the initial stages, a good screening program is essential for the early detection and treatment of this condition. Description of the various stages and threshold ROP has been included in the protocol. Guidelines regarding the procedure of dilatation, ophthalmic examination and treatment (if required) has been provided in the protocols. Close cooperation between the ophthalmologist and neonatologist is essential for a successful program. PMID- 11876128 TI - Acute liver failure. AB - Acute liver failure (ALF) is a rare condition in the pediatric population. Patients who present with severe failure of liver synthetic function have a high mortality with medical therapy alone. The main causes of death are cerebral edema, hemorrhage, renal failure and sepsis. The etiology of ALF is age specific, with a significant number due to inborn errors of metabolism especially in neonates and infants. Treatment of children with ALF is supportive, aimed at preventing and managing associated complications until the native liver recovers or liver transplantation. Sedation should not be administered unless a decision for artificial ventilation has been made. As all children are potential transplant candidates, transfer to and management in a liver transplant centre is recommended. Prognostic criteria for mortality are less well defined compared to the adult population, although a significantly elevated INR > or = 4 carries a high chance of death, and liver transplantation should be considered at this stage. Auxiliary transplantation is an attractive option in selected individuals and provides the chance to stop immunosuppression should sufficient hepatic regeneration occur. The use of various liver assist devices and hepatocyte transplantation as a bridge to liver transplantation show promise, although when used in isolation, they do not have an impact on overall patient survival. PMID- 11876129 TI - Autoimmune liver disease. AB - Autoimmune liver disorders are inflammatory liver diseases characterised histologically by a dense mononuclear cell infiltrate in the portal tract and serologically by the presence of non-organ and liver specific autoantibodies and increased levels of immunoglobulin G (IgG), in the absence of a known etiology. They usually respond to immunosuppressive treatment, which should be instituted as soon as diagnosis is made. The onset is variable and often mimics acute hepatitis. The previously accepted requirement of six month duration of symptoms before a diagnosis of autoimmune disease could be made has been abandoned. PMID- 11876130 TI - Adrenal hemorrhage due to vitamin-K deficiency. AB - Adrenal hemorrhage has many etiologies including bleeding diathesis. A 10-day-old female baby was admitted to our clinic with the complaint of abdominal distention. Hemorrhage was determined on the right adrenal gland with abdominal ultrasonography and computerized tomography. Laboratory investigations showed PT 44 sec and aPTT longer than one minute. This article reports here an infant diagnosed as adrenal hemorrhage due to Vitamin-K deficiency presenting as an abdominal mass. PMID- 11876131 TI - [Nothing better--no competition]. PMID- 11876132 TI - [Alcohol and cardiovascular disease]. PMID- 11876133 TI - [Diagnosis and vitamin B12 deficiency]. PMID- 11876134 TI - [Breast feeding in Tromso before and after the baby-friendly-hospital initiative]. AB - BACKGROUND: "The-Baby-Friendly Hospital Initiative" was introduced in the early 1990s by WHO/UNICEF to reverse a declining trend in breastfeeding worldwide. We wanted to investigate factors influencing breastfeeding and whether this initiative, introduced between 1993 and 1996, had improved breastfeeding in our region, Tromso, a mid-sized city in northern Norway. MATERIAL AND METHODS: Medical records at maternal and child health centres of 1,374 infants born 1992 (n = 653) and 1997 (n = 721) were studied. The number of mothers breastfeeding, duration of lactation, parents' age, their occupation and education, maternal marital status and parity were registered. RESULTS: In a multiple regression analysis, birth, year 1997, high parental level of education/occupational prestige, and higher maternal age significantly prolonged the total period of breastfeeding. Parental age, education/occupation, the number of women starting breastfeeding, the duration of exclusive breastfeeding and the total lactation period significantly increased from 1992 to 1997. When correcting for parental age and education/occupation in a multiple regression analysis we found an increase in exclusive breastfeeding and the total lactation period by 0.5 month (mean (95% CI)) 0.5 (0.2-0.8) and 1.1 month (0.6-1.5) respectively. INTERPRETATION: This improvement might be due to "the Baby-Friendly Hospital Initiative", but also to unknown factors. PMID- 11876135 TI - [Significance of maternal alloantibodies for neonatal thrombocytopenia]. AB - BACKGROUND: Thrombocytopenia has been shown to be present in about 1% of unselected newborns. Alloantibodies to platelets induce the most severe thrombocytopenia. MATERIAL AND METHODS: Samples from 195 mothers who had just given birth to thrombocytopenic children, were analysed by platelet antigen genotyping and detection of platelet specific antibodies. RESULTS: 75 mothers were typed human platelet antigen (HPA) 1bb, and in 65 mother, anti-HPA 1a antibodies could be detected. Anti-HPA 5b antibodies were detected in three samples and anti-HLA antibodies in 73 samples. INTERPRETATION: Alloantibodies were shown to be an important cause of thrombocytopenia in the new-born children and anti-HPA 1a antibodies were, as expected, the most common platelet-specific antibody involved. Anti-HLA class 1 antibodies were detected as the only antibody in 51 cases of thrombocytopenia. Though it is not yet formally shown, this may indicate that anti-HLA class 1 antibodies may cause thrombocytopenia in the fetus and new-born. Based on the assumption that neonatal alloimmune thrombocytopenia is present in 1:1,000 new-born, 25% of the neonatal alloimmune thrombocytopenia cases in Norway are verified by laboratory analysis. Alloantibodies to thrombocytes are of clinical importance in future pregnancies and transfusions. The cost and benefit of a national screening program for anti-HPA 1a antibodies in all pregnant women should be carefully considered. PMID- 11876136 TI - [Laparoscopic examination of non-palpable testis]. AB - BACKGROUND: Approximately 20% of undescended testes are impalpable. They can be located intraabdominally, in the inguinal canal, in ectopic position, or they can be missing. Traditionally, surgical groin exploration has been carried out in such cases, sometimes followed by laparotomy if no testis or spermatic cord could be found in the groin. This approach often involves unnecessary and sometimes too extensive surgery. Laparoscopy facilitates the identification of the testicle or proves the lack of it in a less invasive way. MATERIAL AND METHODS: From 1986 to 1999, 77 boys aged one year five months to 13 years 7 months underwent laparoscopy for unilateral or bilateral nonpalpable testis in our department. RESULTS: In 72 boys (94%), the findings were conclusive in the form of intraabdominal testis in 26, an intact intraabdominal spermatic cord entering the deep inguinal ring in 27, and a missing testis in 19 boys. INTERPRETATION: Laparoscopy identified the boys who did not require further surgery because the testis was missing, Furthermore, when a testis was found, laparoscopy was helpful in choosing the most appropriate method of orchiopexy. It is concluded that at present, laparoscopy is the preferred method of investigation in boys older than one year of age with nonpalpable testis. PMID- 11876137 TI - [Acute pyelonephritis in children with minimal or normal urine findings]. AB - BACKGROUND: Over the past years we have seen several children with serious bacterial infections in whom the focus of the infection was found in the kidneys despite of normal or minimal urine findings. MATERIAL AND METHODS: We review the cases of all children hospitalised at our paediatric department from 1995-2000 with normal or minimal urine findings, and in whom acute pyelonephritis was diagnosed by means of computerized tomographic imaging of the kidneys. RESULTS: All 10 children (aged 9 months to 9 years) had focal areas of decreased enhancement in the kidney parenchyma. All received intravenous antibiotic treatment. Vesicouretheral reflux was detected in four patients and required surgical intervention in one. One patient had considerable renal scarring and decreased renal function on dimercaptosuccinic acid (DMSA) scintigraphy. INTERPRETATION: In children with suspected serious bacterial infection, it is important to have the possibility of renal infection in mind in spite of normal urine findings. The importance of establishing the right diagnosis lies in the fact that patients with pyelonephritis need further investigation and follow-up. PMID- 11876138 TI - [Small foreign bodies in the lungs of children--treatment or observation?]. AB - The diagnosis of foreign body aspiration may easily be missed. Three children younger than three years of age inhaled, respectively, popcorn, a plastic pearl, and a piece of tape to the lungs. The children were admitted to the paediatric department under strong clinical suspicion of foreign body inhalation, but were discharged with minor symptoms and after normal X-ray investigation. Two of the children expelled their foreign body (tape and popcorn) after three days and three weeks, respectively, whereas the plastic pearl was removed by bronchoscopy after six months. X-ray investigation of the lungs are of limited value in the exclusion of presence of small inhaled foreign bodies. Children with minor symptoms suspected of having inhaled foreign bodies may possibly be observed for a short period in order to see if the foreign body is expelled spontaneously. However, the children should be carefully followed up, and bronchoscopy should be considered at all times. PMID- 11876139 TI - [Stickler's syndrome--an underdiagnosed condition?]. AB - BACKGROUND: Stickler's syndrome is an autosomal dominantly inherited connective tissue disorder characterised by ocular, orofacial, skeletal and auditory features. The estimated prevalence is 1:10,000. MATERIAL AND METHODS: We present a girl with the salient features of Stickler syndrome. Based on a literature search on Medline, we present an overview of this disorder. RESULTS: The patient presented at birth with Pierre Robin sequence and bilateral exophtalmus. Serial ophthalmological investigations have revealed a non-progressive myopia of high degree and abnormalities of the vitreous gel architecture. From the age of three, she had joint hypermobility and joint pain. Her intelligence is normal, but she requires speech therapy because of problems with articulation. INTERPRETATION: Recent research has provided a better understanding of the molecular genetic background of this condition. According to mutations in three genes encoding type II- and/or type XI-collagen, Stickler's syndrome can be subclassified into type 1, 2 and 3, but there is a considerable clinical overlap in symptoms. Patients with mild symptoms may be undiagnosed. Once the diagnosis is established, a coordinated multidisciplinary follow-up approach is recommended. PMID- 11876140 TI - [DiGeorge syndrome. An underdiagnosed disease category with different clinical features]. AB - BACKGROUND: DiGeorge syndrome is estimated to affect one in every 3,000-4,000 live-born individuals. The syndrome is also known as velocardiofacial syndrome (VCFS) and conotruncal anomaly face syndrome (CTFS). The most common clinical features are mental retardation, congenital heart anomalies, primary hypoparathyroidism (with hypocalcaemia), aplasia or hypoplasia of the thymus, and a dysmorphic face. 90% of the affected individuals have a deletion at the long arm of chromosome 22. 80-90% of those deletions are de novo mutations. MATERIAL AND METHODS: This article presents the case of a 32-year-old woman who was diagnosed with DiGeorge syndrome after a grand mal seizure due to hypocalcaemia. The hypocalcaemia was caused by primary hypoparathyroidism. We also give a brief review of the current state of knowledge about DiGeorge syndrome. RESULTS AND INTERPRETATION: DiGeorge syndrome is probably underdiagnosed. A correct and early diagnosis is important in order to prevent medical complications, e.g. hypocalcaemia and hypothyrosis, and to evaluate the patient's overall need of care. PMID- 11876141 TI - [Leflunomide--a new disease modifying anti-rheumatic agent]. AB - BACKGROUND: Leflunomide is a novel disease-modifying antirheumatic drug (DMARD) for treatment of rheumatoid arthritis. The agent has been developed for the treatment of rheumatoid arthritis, but its multiple immunomodulatory properties may in the future be of interest in the treatment of other rheumatic and immunological diseases. MATERIAL AND METHODS: Review of the literature in order to present the current relevant clinical documentation of the drug. RESULTS: The clinical documentation is mainly based on three large, prospective, randomized trials of six months "to two years" duration comparing leflunomide with placebo, sulphasalazine or methotrexate. The efficacy of leflunomide in all trials was superior to placebo and comparable to sulphasalazine and methotrexate. The frequency of adverse events was also comparable to the comparators. INTERPRETATION: Leflunomide is a safe and efficacious addition to the roster of antirheumatic drugs, but further clinical trials and experience from clinical practice are needed in the evaluation of its place as a disease-modifying agent. PMID- 11876142 TI - [Incidents connected to pregnancy and labor]. AB - In the period 1993-2000 the Norwegian Board of Health investigated 70 cases of claimed professional misconduct in relation to pregnancy or delivery. There were two maternal deaths, 38 perinatal deaths and 17 cases of cerebral damage. The Board of Health found reason to criticize midwives in 11 cases, obstetricians in 39 cases, and the hospital for lack of routines in 18 cases. Many of the mishaps were connected to false interpretation of fetal signs of distress (clinical signs or cardiotocography), and the investigation concluded that Caesarean section was delayed too long. The number of patient complaints has been constant through the period, in contrast to the widespread belief that misconduct and complaints are steadily increasing. The health authorities and the Norwegian Association of Gynaecologists have focused on quality and standards in obstetrics through committees for investigation of perinatal deaths, national guidelines in obstetrics, and improved national statistics. PMID- 11876143 TI - [COX-2 inhibitors--Columbus' egg or emperor's new clothes?]. PMID- 11876144 TI - [COX-2 inhibitors and cardiovascular adverse effects]. PMID- 11876145 TI - [Natural antimicrobial peptides--promising new antibiotics or a problem later on?]. AB - BACKGROUND: Antimicrobial proteins and peptides are important elements of host defence that are found in virtually all living species examined so far. MATERIAL AND METHODS: This review is based on the author's own research and a search of Medline (key words: antimicrobial peptides, biochemistry, mode of action, resistance). RESULTS: In animals, antimicrobial peptides are found on mucosal epithelial surfaces, in body fluids and in the microbicidal organelles of phagocytic cells. The peptides vary in size, structure and activity, but most are amphiphilic and positively charged. They act on a broad range of bacteria, fungi, viruses, parasites and certain tumour cells. Cationic peptides can be classified into several groups on the basis of sequence similarities, secondary and tertiary structure, function and origin. Their main acting mechanism is thought to be the disruption of the cytoplasmic membrane which kill bacteria swiftly and thoroughly. INTERPRETATION: Antimicrobial peptides are promising novel antibiotic candidates for clinical use, topically as well as systemically. So far, development of resistance against antimicrobial peptides has not been a problem, but this will have to be monitored closely in the years ahead. PMID- 11876146 TI - [Bacteria-killing viruses, Stalinists and "superbugs"]. AB - In June 2000, the WHO warned that the level of resistance to drugs used to treat common infectious diseases is now reaching a crisis point. If world governments do not control infections better in order to slow down the development of drug resistance, entire populations could be wiped out by superbugs against which there is no efficient treatment. Development of resistance is due to both underuse and overuse of drugs, and strategies have been worked out, to slow down the development of resistance for instance by the Norwegian Ministry of Health and Social Affairs. The present article deals with an old principle, mainly developed behind the Iron Curtain, which is now attracting renewed attention in the west: the application of bacterial viruses (bacteriophages) in the fight against bacteria. According to clinical trials in Eastern Europe, mostly uncontrolled, phages have been used successfully in treatments against antibiotic resistant bacteria, for instance in suppurative wound infections, gastroenteritis, sepsis, osteomyelitis and pneumonia. These encouraging data are supported by recent findings in well-controlled animal models demonstrating that phages can rescue animals from a variety of fatal infections. The present review discusses possible advantages and limitations of phage treatment in humans. PMID- 11876148 TI - [Obstetric departments, delivery units and births in Norway in the 1990s]. PMID- 11876149 TI - [Chronic pain--is everything we do wrong?]. PMID- 11876147 TI - [Can disinfectants contribute to antibiotic resistance?]. AB - BACKGROUND: Disinfectants are widely used in medicine, veterinary medicine, and the food processing industry. Increasingly, disinfectants are included in consumer products. Broad-scale use of antiseptics and disinfectants may have detrimental ecological consequences, for instance the development of antimicrobial resistance. MATERIAL AND METHODS: We give an overview of the correlation between the use of certain antiseptics and disinfectants, bacterial resistance to these agents, and antibiotic resistance. RESULTS: The mechanisms of antibiotic and biocide resistance share many common characteristics. There are links between disinfectant resistance and antibiotic resistance. Some biocides have the ability to select for antibiotic resistant mutants and vice versa. Resistance genes are often located on transferable genetic elements that facilitate horizontal gene transfer between microorganisms. Antibiotic resistance and disinfectant resistance may be stabilized and maintained even in the absence of a direct selective pressure. Higher incidence of bacteria resistant to certain disinfectants have been reported in environments where such agents are frequently used compared to environments where they are not in regular use. Increased domestic usage of non-antibiotic antimicrobial agents may select for antibiotic resistant bacteria of clinical significance. INTERPRETATION: The use of antiseptics and disinfectants should be restricted to products and areas where they have an essential and documented effect. PMID- 11876150 TI - [Young women and alcoholic intoxication]. PMID- 11876151 TI - [Sports diving--safety equipment exchanged for instruction]. PMID- 11876152 TI - [Tinnitus]. PMID- 11876153 TI - [Hitler's personality--one more time]. PMID- 11876154 TI - Commentary on Misconceptions and misuse of the MMPI-2 in assessing personal injury claimants. PMID- 11876155 TI - Commentary on Psychometric concerns in neuropsychological testing. PMID- 11876156 TI - Comment on article "Does pain confound interpretation of neuropsychological test results?". PMID- 11876157 TI - Comment on Faulder Colby "Using the binomial distribution to assess effort: forced-choice testing in neuropsychological settings". PMID- 11876159 TI - Resource Directory 2002. Need-to-know organizations & contacts at your fingertips. PMID- 11876158 TI - Improvements in mobility for people with disabilities. PMID- 11876160 TI - Evaluation of capillary perfusion. PMID- 11876161 TI - ["Licence" requirements for physicians who are to perform laparoscopic surgery! Frequency of complications still too high]. PMID- 11876162 TI - Current and ongoing clinical development of immunotherapy in B-cell malignancies. Proceedings of a conference. January 2001, Kailua-Kona, Hawaii, USA. PMID- 11876164 TI - Medical errors summit held in September. PMID- 11876163 TI - COX-2 specific inhibitor, NS-398, increases macrophage migration inhibitory factor expression and induces neuroendocrine differentiation in C4-2b prostate cancer cells. AB - BACKGROUND: Cyclooxygenase-2 (COX-2) is an inducible enzyme involved in the conversion of arachadonic acid to prostaglandins and other eicosaniods. Persistent COX-2 expression is associated with multiple forms of cancer.Therefore, there is much interest in COX-2 specific, non-steroidal anti inflammatory drug use for cancer chemotherapy. The mechanism by which these drugs inhibit tumor growth and progression is unclear, and our knowledge about their potential to prevent or treat prostate cancer is inadequate. MATERIALS AND METHODS: The effects of NS-398, a selective COX-2 inhibitor, on human prostate carcinoma cell line LNCaP and the LNCaP subline C4-2b were investigated in this study. NS-398 effects on apoptosis were examined by caspase-3 activity increase, as well as internucleosomal cleavage. ELISA and PCR were used to determine inhibitor effects on macrophage migration inhibitory factor (MIF) and COX-2 production. RESULTS: At 10 microM, NS-398 treatment resulted in increased production of COX-2 and the pro-inflammatory cytokine, MIF by the C4-2b LNCaP subline. NS-398 (10 microM) induces apoptosis in LNCaP cells, but not in the more aggressive, androgen-unresponsive C4-2b cells. The C4-2b cells were observed to continue to proliferate when treated with NS-398 and continued to retain malignant phenotype characteristics. NS-398 treatment resulted in C4-2b cell differentiation into an unusual neuroendocrine-like cell. These neuroendocrine like cells produced both epithelial (cytokeratin 18 and prostate specific antigen) and neuronal (neuron-specific enolase and chromogranin A) proteins. Furthermore, this C4-2b cellular response to NS-398 was mediated by NF-kappa beta transcription factor activation. CONCLUSIONS: These data suggest that COX-2 inhibition induces NF-kappa beta transcription factor activation, which subsequently induces pro-inflammatory protein expression (COX-2 and MIF) and neuroendocrine differentiation in the LNCaP C4-2b subline. These data provide further evidence that pro-inflammatory protein expression may play an important role in prostate cancer progression. PMID- 11876165 TI - Nurses' knowledge untapped by consumers. PMID- 11876166 TI - California to mandate nurse-patient staffing ratio. PMID- 11876167 TI - Does taking an orthostatic blood pressure include taking the pulse? PMID- 11876168 TI - A randomized, double-blind placebo-controlled trial of a Bach Flower Remedy. PMID- 11876169 TI - Acute coronary syndromes: new developments in pharmacological treatment strategies. PMID- 11876170 TI - The declining comprehensiveness of primary care. AB - BACKGROUND: Recent studies suggest that comprehensiveness of primary care has declined steadily over the past decade. This study tracks the participation rates of general practitioners and family physicians in 6 nonoffice settings across Ontario and examines among which types of physicians this decline in comprehensiveness has occurred. METHODS: Billing (claims) records were used to determine the proportions of fee-for-service general practitioners and family physicians who provided emergency, inpatient, nursing home, house call, anesthesia or obstetrical services from 1989/90 to 1999/2000. "Office-only" physicians were those who worked in none of these nonoffice settings. The relation of various physician characteristics to comprehensiveness of care was tested with multivariate analysis for 1999/2000. RESULTS: The proportion of "office-only" general practitioners and family physicians rose from 14% in 1989/90 to 24% in 1999/2000 (p < 0.001). Significant increases in this proportion were noted among general practitioners and family physicians of all ages, both sexes and all practice locations. In 1999/2000, recent graduates (who had completed medical school within the past 7 years) had higher participation rates for emergency medicine (40% v. 5% for physicians aged 65 years and older); female physicians had higher participation rates for obstetrics (16% v. 11% for males); and older physicians had higher participation rates for nursing home visits and house calls (20% and 57% respectively v. 11% and 37% for recent graduates). However, "office-only" physicians were more likely to be female (odds ratio [OR] 2.65, 95% confidence interval [CI] 2.37-2.96), recent graduates (OR 1.35, 95% CI 1.15-1.60), aged 65 years and older (OR 1.45, 95% CI 1.20-1.75) or practising in a city with a medical school (OR 2.30, 95% CI 2.06-2.56) and were less likely to be rural physicians (OR 0.31, 95% CI 0.24-0.41) or certified in family medicine (OR 0.58, 95% CI 0.52-0.66). INTERPRETATION: There has been a decline in the provision of comprehensive care by general practitioners and family physicians in Ontario. The decline is evident across all age groups and for both male and female physicians. It is also evident in rural areas and in cities with and without medical schools. PMID- 11876171 TI - Global history of science comes of age. [Review of: McClellan, JE 3d; Dorn, H. Science and technology in world history: an introduction. Johns Hopkins University Press, 1999]. PMID- 11876172 TI - One stop clinics should not be abandoned. PMID- 11876173 TI - Performance league tables. Use of language should be more careful in describing league tables. PMID- 11876174 TI - Performance league tables. Dr Foster's ranking of hospitals in good birth guide is misleading. PMID- 11876175 TI - Performance league tables. NHS is national but not uniform. PMID- 11876176 TI - Performance league tables. Publication of league tables needs to be open and accurate. PMID- 11876177 TI - Effect of patient centredness and positive approach. Partnership of patient and doctor may provide key to patient satisfaction. PMID- 11876178 TI - Somatisation in primary care. Descriptive use of term should not be confused with its conceptualisation. PMID- 11876179 TI - Quality of Cochrane reviews. Another study found that most Cochrane reviews are of a good standard. PMID- 11876180 TI - Demand for prostate specific antigen testing in primary care. Can the demand for PSA testing in primary care be managed? PMID- 11876181 TI - Symposium. Capacity building for research in minority health. Introduction. PMID- 11876182 TI - Capacity for research in minority health: the need for infrastructure plus will. AB - Cardiovascular mortality has continued to decline, but racial disparities in cardiovascular diseases (CVD) continue to grow. To build the capacity to address these racial disparities, two things will be required. First, a research and policy infrastructure must be in place to provide guidance on what to do and how to do it. Second, the will to implement and activate this infrastructure must be present at the community and policy-making levels. The Jackson Heart Study is an example of a research infrastructure with the economic resources, scientific expertise, and technical manpower required to monitor, organize, assess, and follow a cohort of individuals over time to study the burden, natural history, predictive factors, and level of care for CVD in an African American community. The creation of will within the community for CVD research may require additional strategies than in the majority community, such as community organization and local policy development. These additional efforts at the community level should create a fertile environment to develop research and, ultimately, test strategies for reducing national disparities in cardiovascular health. PMID- 11876183 TI - History of medical ethics and perspectives on disparities in minority recruitment and involvement in health research. AB - The legitimate and successful recruitment of minorities as research participants in clinical trials should be addressed from an ethical and historical perspective. To gain an appreciation of the challenges, to develop strategies and to overcome the disparities of minority involvement in clinic trials, it is essential to be cognizant of previous violations and abuses of ethics and human rights. Also significant are major legislation, regulations and federal initiatives that resulted from those abuses. From history, we have learned we cannot generalize data and assume that, if we have the majority group in clinical trials, then we can accurately apply that data to minorities. There are cultural and environmental differences; thus, it is absolutely crucial that researchers approach recruitment of minority groups with cultural competence and cultural sensitivity. Federal regulations and legislation set the framework for protection of human participants in research. PMID- 11876184 TI - Building capacity for health disparity research at minority institutions. AB - The science and technology enterprise of the United States has consistently produced seminal work and cutting-edge technologies. It has responded promptly to both new opportunities and urgent crises. The success of this enterprise derives largely from the diversity of the types of institutions doing the work and from the many sources of public and private funding available to accomplish it. To those who argue that public-sector funds should support only the best science at the premier research institutions on the nation's East and West coasts, Dr. Rita Colwell, the director of the National Science Foundation (NSF) eloquently responds, "No one region, no one group of institutions, and no special communities have a corner on the market of good and great ideas, smart people, or outstanding researchers. Great ideas can come from just about anywhere." PMID- 11876185 TI - Ethical challenges in community-based research. AB - Investigators in population-based studies confront unique ethical challenges due to the community context of their research, their methods of inquiry, and the implications of their findings for social groups. Issues surrounding requirements for informed consent, the protection of privacy and confidentiality, and relationships between investigators and participants take on greater complexity and have significance beyond the individual research subject. In this paper, ethical challenges associated with community-based epidemiological research are briefly examined. We argue that ethically responsible population-based studies must seriously consider community needs and priorities and that researchers should work collaboratively with local populations to implement study goals. Strategies that promote respect for populations in community-based studies are outlined. These include community participation in research development, implementation and interpretation; adequate provision of information about study objectives to community members; and systematic feedback of study results. PMID- 11876186 TI - Research capacity and closing the health gap. AB - In January 2000, the Healthy People 2010 initiative was introduced by the US Department of Health and Human Services (DHHS), which set the nation's health goals for the coming decade. In 1990, a previous DHHS administration had released the Healthy People 2000 initiative, which set goals to be met by the year 2000. In 1979, the first set of health goals for the nation was released, titled Healthy People. Thus, Healthy People 2010 is the third iteration of such goals. This effort has achieved momentum and indeed is paying off handsomely. The framework of the Healthy People initiatives, however, is just one gauge of US public health care status; it and a myriad of other programs and studies in recent decades reflect the gains and shortcomings of our complex health care system. In several areas, minority health disparities continue to be the weakness in our capacity building to achieve the nation's health goals. PMID- 11876187 TI - Developing community capacity and improving health in African American communities. AB - Community-based programs have produced mixed results. Community capacity is thought to be a major determinant of program effectiveness. Thus, enhancing community capacity may increase the beneficial effects of existing programs and enhance future program effectiveness. This highlights the need to focus on understanding the components of capacity and the methods of enhancing capacity. Although we are just beginning to examine and understand key concepts, community capacity is probably influenced by both relatively nonmodifiable characteristics (such as demographic factors, institutional resources, and social structures) and relatively modifiable characteristics (such as knowledge, skills, and the ability and willingness of members and agencies to work collaboratively). In their relationships with community members and agencies, academicians and public health practitioners may help acquire categorical funding to enhance opportunities to build community capacity and their own capacity as well. The relationship between academicians/practitioners and community members/agencies probably is influenced by a host of characteristics which determine the degree to which capacity can be built. This paper discusses: the key components of capacity; the factors that influence building capacity through collaborations; a community health advisor (CHA) model which both builds on sociocultural aspects of African American culture and is consistent with methods for building community capacity; and how modifications to this model allow it to be compatible with categorically funded projects. PMID- 11876188 TI - Building a gateway to promote cardiovascular health research in African American communities: lessons and findings from the field. AB - African American communities traditionally mistrust academic research. This forms a significant barrier to understanding cardiovascular risk factors in this population, which bears an excess risk of cardiovascular disease and stroke. A clergy/academic partnership was established to build a gateway for salient research and for improving resources for reducing cardiovascular disease risk in the community. From this partnership emanated the African American Family Heart Study. People with a family history of premature coronary heart disease (CHD) have an increased risk for the disease--as high as 12 times that of the general population, if among siblings. Considerably less is known about the actual remediable risk factors in African American families with premature CHD. We initiated the Family Heart Study with a full characterization of 161 apparently healthy, unaffected 30- to 59-year-old African Americans whose siblings were 85 African American index cases with documented premature CHD prior to 60 years of age. We compared their risk factor values to population reference norms obtained in the Third National Health and Nutrition Examination Survey (NHANES III) and the National Health Interview Survey (NHIS) for cigarette smoking. Only 13% of African American male siblings and 14% of female siblings from these families were without any major remediable risk factors. The fact that so many siblings were at extremely high risk calls into question the current applications by provider systems of national guidelines in high-risk African American families. This is an easily identifiable population that would be likely to benefit greatly from targeted screening and culturally sensitive and appropriate treatment. PMID- 11876189 TI - The Strong Heart Study: interaction with and benefit to American Indian communities. AB - The Strong Heart Study (SHS) is a large, multi-community study to investigate the high incidence of cardiovascular disease (CVD) among American Indians and to improve long-term health outcomes within the communities. CVD is the No. 1 cause of death for three geographically and culturally diverse American Indian populations. The SHS investigated the prevalence and incidence of CVD as well as the distribution of risk factors in members of 13 American Indian tribes in Oklahoma, Arizona, and (as one region) North and South Dakota. The SHS enrolled more than 4500 participants ranging in age from 45 to 74 years. During the three phase study, which began with baseline examinations from 1989 through 1991 and concluded in 1999, participants received medical examinations in each phase. To build trust and cooperation, SHS investigators worked closely with tribal leaders and community members, including establishing empowered committees and hiring community members for jobs within the study. As a result of such cooperation, the SHS maintained retention rates of about 90% across all three phases. Benefits to tribal communities also included health care improvements and education for healthy lifestyles. The SHS could serve as a model for similar studies of American Indians or other minority groups. PMID- 11876190 TI - Focus group data pertinent to the prevention of obesity in African Americans. AB - The increased prevalence of obesity among African-American women makes it likely that they bear a disproportionate burden of comorbidities attributable to obesity, such as diabetes, hypertension, and hyperlipemia. These observations suggest that intensive efforts to prevent obesity should be directed at this group. This presentation provides a summary of the findings of focus groups that convened prior to the Sisters Together, Move More, Eat Better campaign in Boston. This pilot campaign was designed to increase awareness of the importance of healthy eating and physical activity among young adult African-American women. In addition, data collected by the Division of Nutrition and Physical Activity of the US Centers for Disease Control and Prevention (CDC) provide additional information about attitudes toward diet and physical activity among African American youth. Such data are essential to understand the attitudinal changes necessary to prevent obesity in these vulnerable populations. PMID- 11876191 TI - Programs of the Robert Wood Johnson Foundation to develop minority medical careers. AB - The Robert Wood Johnson Foundation (RWJF) is the nation's largest philanthropy devoted to health. The foundation long has been concerned about increasing diversity in the health professions. Between 1972 and 1981, grants totaling nearly $6.7 million were made to medical, medical/dental schools, or other educational organizations to support minority students. Funds enabled students who were interested in applying to medical or dental school to enroll in special preparatory courses. Most students were African American. One program, however, targeted US Puerto Rican students and other Hispanic students. Nearly 2500 students enrolled in these preapplication enrichment programs. Data reported to the foundation on medical or dental school acceptance for 1959 of these students indicated that 57% of students were successful. An additional $10.5 million in grants were awarded during this period: $2.5 million to provide scholarships for minority group medical students, $580,000 to support preceptorships with minority physicians/mentors, and $7.5 million to strengthen Meharry Medical College's Comprehensive Primary Care Health Science Program. In the early 1980s, the RWJF Board of Trustees considered a series of staff analyses, which resulted in additional direct support to historically black medical schools, including Meharry and those at Drew University and Morehouse College. These analyses also set the stage for two RWJF programs, the Minority Medical Faculty Development Program and the Minority Medical Education Program, which exist today. This article describes these programs, along with the more recent Health Professions Partners Initiative, and offers reflection and analysis about their impact on diversity in the medical profession. PMID- 11876192 TI - Bogalusa Heart Study: a long-term community study of a rural biracial (Black/White) population. AB - The Bogalusa Heart Study, a long-term population study with a continued relationship with a community, addresses the problem of capacity building in minority health research. The study was originally funded as a Specialized Center of Research-Arteriosclerosis (SCOR-A) by the National Heart Lung and Blood Institute (NHLBI). These centers were to conduct research on atherosclerosis, coronary artery disease (CAD), hypertension, diabetes mellitus, and complications of cardiovascular-renal disease as the major causes of deaths in the United States. From earlier research on atherosclerosis, we became interested in the underlying characteristics in early life that would eventually lead to clinical morbidity and mortality from heart disease. An observation at autopsy showed the degree of atherosclerotic involvement in human aortas, from young to older individuals (Figure 1). For example, at age 40 years, marked individual variability occurred in the severity and involvement with atherosclerotic disease. Some individuals showed very little disease, while almost 70% of the surface was diseased in others. Further studies on arterial wall matrix showed aortas from young individuals varied with the extent of disease and its chemical composition. This background stimulated an interest in studying children for early clinical evidence of major adult heart diseases. The Bogalusa Heart Study was begun in 1972 as an epidemiology study of cardiovascular risk factors in children and adolescents; it eventually evolved into observations of young adults. Bogalusa, LA, is a biracial (black/white) rural community 70 miles north of New Orleans, comparable to many other communities in southeastern United States. PMID- 11876193 TI - Potential applications of differential permeability membranes in the control of gas composition in closed circuit atmospheres. AB - Long hyperbaric exposures of humans, at pressures in excess of 2.5 atmospheres, require that the oxygen concentration of the breathing gas be maintained at levels less than atmospheric air, in order to prevent pulmonary oxygen toxicity. Carbon dioxide produced by humans must be removed from the breathing gas of closed environments. Differential permeability gas separation devices can be used to reduce the concentrations of carbon dioxide and oxygen in recirculating nitrogen/oxygen atmospheres. The application of these devices can eliminate the requirement for nitrogen to dilute oxygen, and reduce, or eliminate, the requirement for carbon dioxide absorbents. The use of differential permeability gas separation devices for the production of nitrogen/oxygen breathing gas mixtures for divers, using air as a source gas, can produce mixtures with unacceptably high levels of carbon dioxide. PMID- 11876195 TI - Proposed minimum requirements for the operational characteristics and testing of submersible atmosphere monitoring and control units. AB - The design of a submersible and its myriad subsystems is a complex undertaking made more difficult by the interval between new designs. Special applications also tax the life support designer, such as diver lock-out submersibles, which are far more complex than one-atmosphere vehicles. The long intervals between designs generally means that a new generation of designers will be approaching the problems without the benefit of the knowledge and experience of the past generation of designers. This new generation of designers often has greatly improved technologies and materials at hand. Yet in the application of the new technology, life support issues that were addressed and resolved in the old and proven designs get lost in the process of redesign due to the lack of experience in the life support arena. Competent engineers operating outside their normal area of expertise can generate costly or even deadly mistakes. For this reason a set of performance requirements that details the critical functions of the life support systems is needed. Atmosphere control electronics can seriously impact the safety of the submersible crew. The triple dangers of fire, toxicity, and asphyxia demand that the electronics that monitor and control the submarine compartment atmosphere have certain characteristics. These characteristics are easily captured in a performance requirement. This article will present a set of proposed minimum performance requirements, with the goal of establishing a dialog for the creation of guidelines for the classification, rating, design, and testing of embedded electronics for life support systems used in submersible applications. These guidelines will serve as the foundation for the later creation of a set of industry specifications. PMID- 11876194 TI - Current submarine atmosphere control technology. AB - Air purification in submarines was introduced towards the end of World War II and was limited to the use of soda lime for the removal of carbon dioxide and oxygen candles for the regeneration of oxygen. The next major advances came with the advent of nuclear-powered submarines. These included the development of regenerative and, sometimes, energy-intensive processes for comprehensive atmosphere revitalization. With the present development of conventional submarines using air-independent propulsion there is a requirement for air purification similar to that of the nuclear-powered submarines but it is constrained by limited power and space. Some progress has been made in the development of new technology and the adoption of air purification equipment used in the nuclear-powered submarines for this application. PMID- 11876196 TI - Development of prototype full-automatic environmental control system for nitrox saturation diving. AB - A full-automated hyperbaric environmental control system (HECS) for nitrox saturation diving HABITAT has been examined since 1994. HECS was planned to be located inside HABITAT and operated automatically by supplying electricity and cooling water. To realize this, a regenerative-type carbon dioxide removal method and AC 100 V electric powered blower system were developed as key technology. By using molecular sieves as an adsorbent material for carbon dioxide, automated sequence for adsorbent and regeneration could be successfully realized. Also, by using solid-state conductor circuit, an AC 100 V type blower to use under hyperbaric conditions could be developed. A prototype HECS was manufactured and settled in JAMSTEC diving simulator for further evaluation. PMID- 11876197 TI - Aquarius: transitioning to an unmanned life support buoy. AB - The Aquarius underwater laboratory (or habitat) is the world's only operational saturation facility currently supporting scientific research and is operated by the University of North Carolina at Wilmington. The underwater laboratory accommodates and supports six aquanauts (scientists and habitat technicians) at habitat depth for 10-30-day missions. In the past, life support systems were provided by a manned support barge or Mobile Support Base (MSB) moored directly above Aquarius. The MSB was manned 24 h a day during saturation missions, which required 12 support staff in three separate shifts. A new unmanned Life Support Buoy (LSB) replaces the MSB and provides life support systems and is the voice, video, and data communications bridge from the habitat to the shore base. The LSB transmits status of all life support systems to the habitat and the shore base, thus minimizing the need for support staff to be present overtop of Aquarius during missions. PMID- 11876198 TI - Life support in small one-atmosphere underwater work systems. AB - Manned submersibles and atmospheric diving suits share a number of characteristics, with the most obvious being the rigid hull structure required to maintain a one-atmosphere cabin pressure in the presence of high external pressures. The rigid pressure hull results in a fixed internal volume that allow a simple, mechanical means of precisely controlling the input of make up oxygen to replace that metabolized by the occupant(s). The field-proven life support hardware described in this article is peculiar to monobaric systems but may have applications in other manned cabin situations where a fixed volume is present- regardless of maintenance pressure. Also briefly described is a state-of-the-art life support monitoring system developed by Nuytco Research Ltd. under contract to the National Energy Board of Canada. The "Biosensor" is thought to have broad application potential in the manned undersea field. PMID- 11876199 TI - Closed circuit, multiperson, lung-powered rebreathers for hyperbaric oxygen treatment. AB - During the conceptual design of a "fly away" Submarine Rescue System (SRS) for the US Navy, a closed circuit breathing system for hyperbaric oxygen treatment was chosen to reduce the weight and volume of the necessary oxygen stores used in decompression treatments. A multiperson, human-powered breathing system was modeled and analyzed for potential development. The potential development of a prototype will allow empirical testing of performance parameters such as breathing, resistance, oxygen concentration, and hyperbaric gas analysis. As a spinoff of the multiperson concept, the modification of commercial off-the-shelf closed circuit air rebreathers are now being considered as an alternative for development. Each concept has benefits and limitations in providing a solution to the procurement of a key component to the SRS. PMID- 11876200 TI - Microbial contamination of advanced life support (ALS) systems poses a moderate threat to the long-term stability of space-based bioregenerative systems. AB - Microbial contamination of terrestrial hydroponic plant-growing systems provides an effective analogue for studying microbial contamination issues for space-based advanced life support (ALS) systems. If aggressive root or foliar pathogens are introduced into ALS hydroponic systems, severe epidemics are probable. Greater than 80% of the reported outbreaks of root pathogens in terrestrial hydroponic systems describe epidemics caused by fungal species of Fusarium, Phytophthora, and Pythium. However, it is likely that a comprehensive sanitation and quarantine program for space-based ALS modules will prevent contamination by Phytophthora and Pythium spp. because these pathogens are general soilborne. However, Fusarium spp. are typically airborne, can grow saprophytically on diverse substrates, and have been common contaminants of American spacecraft. If comprehensive sanitation and quarantine programs are established for space-based ALS modules, then the threat of pathogen introductions into these systems will be significantly mitigated. Microbial contamination studies in spacecraft over the last 30 years indicate that a high diversity of bacteria, fungi, and actinomycetes are commonly carried on board probably via clothing, equipment, air currents during spacecraft handling and loading, food, and the astronauts themselves. Species of Alternaria, Aspergillus, Botrytis, Candida, Cephalosporium, Cladosporium, Fusarium, Mucor, Penicillium, Phoma, and Trichoderma were the most prevalent fungi recovered, and species of Bacillus, Escherichia, Klebsiella, Micrococcus, Pseudomonas, Staphylococcus, and Streptococcus were the most prevalent bacteria recovered from spacecraft. Most of these genera contain species that have been reported as plant pathogens. Strict quarantine procedures were not effective in preventing contamination of spacecraft during these missions. Research must be initiated to better understand how microorganisms interact with plants and animals in microgravity environments because microbial contamination of spacecraft and ALS modules cannot be avoided. An integrated pest management (IPM) program likely will be developed for managing plant disease outbreaks in space-based ALS systems. PMID- 11876201 TI - Consumer acceptance of vegetarian sweet potato products intended for space missions. AB - Sweet potato is one of the crops selected for NASA's Advanced Life Support Program for potential long-duration lunar/Mars missions. This article presents recipes of products made from sweet potato and determines the consumer acceptability of products containing from 6% to 20% sweet potato on a dry weight basis. These products were developed for use in nutritious and palatable meals for future space explorers. Sensory evaluation (appearance/color, aroma, texture, flavor/taste, and overall acceptability) studies were conducted to determine the consumer acceptability of vegetarian products made with sweet potato using panelists at NASA/Johnson Space Center in Houston, TX. None of these products including the controls, contained any ingredient of animal origin with the exception of sweet potato pie. A 9-point hedonic scale (9 being like extremely and 1 being dislike extremely) was used to evaluate 10 products and compare them to similar commercially available products used as controls. The products tested were pancakes, waffles, tortillas, bread, pie, pound cake, pasta, vegetable patties, doughnuts, and pretzels. All of the products were either liked moderately or liked slightly with the exception of the sweet potato vegetable patties, which were neither liked nor disliked. Mean comparisons of sensory scores of sweet potato recipes and their controls were accomplished by using the Student t-test. Because of their nutritional adequacy and consumer acceptability, these products are being recommended to NASA's Advanced Life Support Program for inclusion in a vegetarian menu plan designed for lunar/Mars space missions. PMID- 11876202 TI - Sweet potato in a vegetarian menu plan for NASA's Advanced Life Support Program. AB - Sweet potato has been selected as one of the crops for NASA's Advanced Life Support Program. Sweet potato primarily provides carbohydrate--an important energy source, beta-carotene, and ascorbic acid to a space diet. This study focuses on menus incorporating two sets of sweet potato recipes developed at Tuskegee University. One set includes recipes for 10 vegetarian products containing fom 6% to 20% sweet potato on a dry weight basis (pancakes, waffles, tortillas, bread, pie, pound cake, pasta, vegetable patties, doughnuts, and pretzels) that have been formulated, subjected to sensory evaluation, and determined to be acceptable. These recipes and the other set of recipes, not tested organoleptically, were substituted in a 10-day vegetarian menu plan developed by the American Institute of Biological Sciences (AIBS) Kennedy Space Center Biomass Processing Technical Panel. At least one recipe containing sweet potato was included in each meal. An analysis of the nutritional quality of this menu compared to the original AIBS menu found improved beta-carotene content (p<0.05). All other nutrients, except vitamin B6, and calories were equal and in some instances greater than those listed for NASA's Controlled Ecological Life Support Systems RDA. These results suggest that sweet potato products can be used successfully in menus developed for space with the added benefit of increased nutrient value and dietary variety. PMID- 11876203 TI - Water purification system by using the biofilter for long-term experiment equipment with aquatic animals for the space station. AB - We have developed a water purification system that enables long-term experiment with aquatic animals for 90 days or more on the space station. We designed the system that combined a biofilter for ammonia removal (nitrification) with another for nitrate removal (denitrification). The experiment with goldfish was for 90 days with an aquatic animals' examination device. The equipment consists of a fish tank, a filter module, pumps, and an artificial lung for gas exchange. The goldfish were kept in the tank without any water replacement throughout the experiment. When a filter module consists of adsorbents without bacteria, the concentration of the nitrite and ammonia begin to increase so that the goldfish die. On the contrary, neither ammonia nor nitrite accumulated throughout the experiment, and the concentration of T-N also maintained 30 ppm or less when the combined biofilter was used. Moreover, no fish died throughout the period. The water purification system with biofilter enabled us to examine the long-term life support testing. We also report a new denitrification (correction of dentrification) method for the life support system. PMID- 11876204 TI - Wetland systems for bioregenerative reclamation of wastewater: from closed systems to developing countries. AB - Results are presented from constructed wetland systems designed to treat wastewater in Akumal, Quintana Roo, Mexico, which was developed after prior experience with the Biosphere 2 closed ecological system wetland systems. These systems illustrate the congruity of needs in advanced life support systems and in solving social and environmental problems in developing countries. For sustainable food production for life support, closed ecological systems need to bioregenerate and recycle nutrient-rich wastewater. Developing countries need low tech ecologically engineered systems that minimize requirements for capital, nonrenewable energy, and technical expertise. Biosphere 2's surface flow wetlands covered 41 m2 and treated the wastewater from eight inhabitants, laboratories, and domestic animals during the 1991-1993 closure experiment. The Mexican wetlands are subsurface flow wetlands using limestone gravel as substrate. Two wetland systems treat sewage from 40 people and cover 131 m2. During the initial year of operation, the wetlands in Akumal reduced BOD 86%, TSS 39%, total P 80%, total N 75%, and coliform bacteria 99.85%. Phosphorus uptake in the limestone gravel was around 6 mg/kg. High biodiversity, with 70 plant species, was maintained in the Akumal constructed wetlands 1.5 years after planting. The Shannon diversity index was 4.7 (base 2). Plant diversity was slightly less than tropical forest ecosystems of the region, but far greater than biodiversity in natural mangrove wetlands. PMID- 11876205 TI - [Social conditions in the reception of scientific knowledge]. AB - Scientific knowledge has recently become dominant in the cognition and interpretation of the world. Its dominance is a result of the competence and authority that science has. The process of the social reception of science has an idealising character. The growing trust in science, on the one hand provides a better opportunity for the rationalisation of behaviour and a better control of natural phenomena, on the other hand it can be a cause of numerous distortions. The transmission of knowledge is a social act as we gain almost all our knowledge from and through other people. In this social process, apart from valid knowledge, mystifications and illusions are also transmitted. PMID- 11876206 TI - [The main field hospitals of the French and Polish armies behind the lines in 1807]. AB - The health requirements of the French and Polish troops fighting in Pomerania, mainly those of General Zajaczek's Observation Corps (Second Legion), forced the interim Polish authorities to establish main field hospitals in the Spring of 1807. Firstly, at Mlawa, and then in the former storehouses on the Vistula at Tokary and Dobrzykow. Though the latter were situated in two localities, they nevertheless constituted an administrative whole. Of temporary character, these large hospitals (Mlawa was capable of accommodating up to 1000 patients, Tokary and Dobrzykow up to 2000 patients) were closed after the end of hostilities. In spite of the temporary character of the field hospitals, the daily death rate was only several persons (sometimes there were a dozen or so fatalities, which also constituted a small number). During the establishment of the main field hospitals, a number of people demonstrated a degree of organisational talent. These included Jozef Wybicki, representative of the Governing Committee and Stanislaw Breza, director of internal affairs. Later there was Rev. Jozef Poniatowski, war commander, also local officials such as Jozef Zablocki and Teodor Mikolaj Dembowski, as well as representatives of the health service such as Dr. Hiacynt Dziarkowski, and Dr. Michal Bergonzoni. Other people worth mentioning include the hospital personnel (Colonel Teodor Pretwitz was Commanding Officer of the field hospitals in Mlawa, Tokary and Dobrzykow) who helped the sick and wounded soldiers, often under the difficult conditions of supplies and staff shortages. These shortages were a result of the scale of the enterprise, the poor relations between staff members, such as the conflict between Dr. Dziarkowski and Colonel Pretwitz, reservations about the competence of the head doctor Juliusz Szamborski. But most of all these shortages were a result of requisitions ordered by French Commissioners and Ward Heads. In spite of the problems main field hospitals achieved their purpose and gained recognition by high-ranking Polish and French commanding officers, especially General Zajaczek. PMID- 11876207 TI - [Carl Gustav Jung's Theatrum Mundi. Can the description of modern alchemy in Jung's Psychologie und Alchemie be really regarded as a historical reconstruction?]. AB - In his work, Psychologie und Alchemie, published in 1944, Jung wanted to present a reconstruction of alchemy. In order to do this he used a method of psychology, which he modified and enriched with inspirations drawn from neo-romantically oriented German medical historiographies of the nineteen thirties. Using historical materials, he intended to demonstrate the empirical character of his Depth Psychology, a widespread concept, ingrained in our supra-personal social unconscious. The present article questions the nature of Jung's reconstruction of alchemy as historical reconstruction. The author presents the methodological bases referred to by Jung derived from his concept of medical practice and compares them with a manner of thinking typical of contemporary historical analysis. The article also presents Jung's inspirations from the medical historiography of his time, as well as the dissimilarity of his concept with the model of historical narrative construction in force in the historiography of the thirties and forties. At the same time, it presents the elements of the "romantic inheritance" in Jung's thought, drawn from the tradition of German non materialistic medicine of the first half of the nineteenth century. PMID- 11876208 TI - Competence assessment approach broadens view of nursing skills. PMID- 11876209 TI - Time for sensitivity training: cultural diversity in cardiovascular disease. PMID- 11876210 TI - Calculating pulmonary artery pressure. PMID- 11876211 TI - Does "ICU psychosis" really exist? AB - In summary, ICU psychosis does not develop in all patients. Instead, many patients are at risk for hypoactive, hyperactive, or mixed hypoactive and hyperactive delirium. Prevention of delirium should always be foremost, including recognition of patients at high risk, minimal use of causative medications, and treatment of physiological conditions that are often unrelated to a patient's admitting diagnosis. When prevention fails, early diagnosis and treatment can make a marked difference in patients' outcomes. The potential adverse outcomes of delirium are well documented. These include increased mortality; increased length of stay; reduced level of functioning in the elderly, which often leads to placement in a nursing home; and stress response syndrome after hospitalization. The value of nursing in preventing delirium is evident when nurses apply their knowledge of potential causes and develop strategies to avoid these causes in their patients. Nurses provide early detection and coordinate with other members of the healthcare team to initiate a plan of care that includes prompt treatment of delirium to reduce the signs and symptoms, duration, and potential adverse sequelae of this disorder. Nursing interventions are designed to enhance patients' cognitive status, sense of security, safety, and comfort. Nurses are instrumental in providing appropriate choices, doses, and administration of medications and in recognizing side effects. Use of medications ordered to treat delirium is often left to nurses' discretion because the orders specify that the drugs should be given as needed. Finally, nurses are the ones who recognize the need for additional assistance via psychiatric consultations or for more intensive observation and management of patients to ensure quality care. PMID- 11876212 TI - Nurse-coached intervention for the families of patients in critical care units. PMID- 11876213 TI - Combining mechanical ventilation with hospice care in the home: death with dignity. PMID- 11876214 TI - Recognizing and managing purple glove syndrome. PMID- 11876215 TI - Nursing care of neonates receiving intravenous prostaglandin E1 therapy who are not in an intensive care unit. PMID- 11876216 TI - Acute acetaminophen overdose in adolescents and adults. PMID- 11876217 TI - Employee competency pathways. PMID- 11876218 TI - Using the Synergy Model to link nursing care to diagnosis-related groups. PMID- 11876219 TI - Alternative and complementary modalities for managing stress and anxiety. PMID- 11876220 TI - What is the current consensus or standard of practice for nurses removing chest tubes in a postoperative setting? PMID- 11876221 TI - Can you clarify the need for gentle versus vigorous bubbling in closed-system chest tube suctioning? PMID- 11876222 TI - True colour imaging of the fundus using a scanning laser ophthalmoscope. AB - Currently retinal imaging is performed with the fundus camera. This has a number of limitations, in particular the high level of illuminations required for imaging. The scanning laser ophthalmoscope (SLO) has been proposed as an alternative imaging device but to date one of its main limitations has been that it gives only monochromatic images. In this paper we describe an SLO which uses low power red, green and blue lasers to image the human fundus. Using three lasers simultaneously to produce a colour image will increase the fundus exposure by a factor of three. To overcome this problem, a technique has been developed for multiplexing the lasers so that each point on the retina is imaged by the three lasers pulsed rapidly in sequence. The total exposure is thus kept to the same level as for a single laser and total imaging time is not increased. An example is shown of the image from a patient with diabetic retinopathy. PMID- 11876223 TI - A multi-shell algorithm to reconstruct EIT images of brain function. AB - Electrical impedance tomography (EIT) may be used to image brain function, but an important consideration is the effect of the highly resistive skull and other extracerebral layers on the flow of injected current. We describe a new reconstruction algorithm, based on a forward solution which models the head as four concentric, spherical shells, with conductivities of the brain, cerebrospinal fluid, skull and scalp. The model predicted that the mean current travelling in the brain in the diametric plane for current injection from polar electrodes was 5.6 times less than if the head was modelled as a homogeneous sphere; this suggests that an algorithm based on this should be more accurate than one based on a homogeneous sphere model. In images reconstructed from computer-simulated data or data from a realistic saline-filled tank containing a real skull, a Perspex rod was localized to within 17% or 20% of the tank diameter of its true position, respectively. Contrary to expectation, the tank images were less accurate than those obtained with a reconstruction algorithm based on a homogeneous sphere. It is not yet clear if the theoretical advantages of this algorithm will yield practical advantages for head EIT imaging; it may be necessary to proceed to more complex algorithms based on numerical models which incorporate realistic head geometry. If so, this analytical forward model and algorithm may be used to validate numerical solutions. PMID- 11876224 TI - Response of some biophysical properties of blood to changes in the perfusion flow rate during cardiopulmonary bypass. AB - Cardiopulmonary bypass (CPB) is a safe support system during major cardiac operations because it allows the distribution of adequately oxygenated blood to the entire body. The optimal perfusion flow to be used during hypothermic CPB remains a controversial issue. In the present study, the effect of different flows and pressures on oxygen delivery, extraction and consumption was compared. Such an effect was evaluated for 40 patients undergoing open-heart surgery using CPB (membrane oxygenator) and systemic hypothermia. The patients were randomized into two groups (20 each) according to the flow and pressure (low flow, low pressure and high flow, high pressure). The electrical impedance from 50 Hz to 10 MHz for blood samples and erythrocytes for both groups was also measured at different time intervals (before-, during and post-CPB). No significant change for low flow group was recorded (p > 0.05) while the dielectric properties of the high-flow group were greatly affected by the time of operation. Hence, low flow can be safely used in young patients without any organ impairment or carotid artery disease, while high flow should be used for patients with any organ dysfunction, at the initiation of CPB and during rewarming to compensate for the increase in oxygen consumption and the need for better perfusion pressure. PMID- 11876225 TI - Optimal imaging with adaptive mesh refinement in electrical impedance tomography. AB - In non-linear electrical impedance tomography the goodness of fit of the trial images is assessed by the well-established statistical chi2 criterion applied to the measured and predicted datasets. Further selection from the range of images that fit the data is effected by imposing an explicit constraint on the form of the image, such as the minimization of the image gradients. In particular, the logarithm of the image gradients is chosen so that conductive and resistive deviations are treated in the same way. In this paper we introduce the idea of adaptive mesh refinement to the 2D problem so that the local scale of the mesh is always matched to the scale of the image structures. This improves the reconstruction resolution so that the image constraint adopted dominates and is not perturbed by the mesh discretization. The avoidance of unnecessary mesh elements optimizes the speed of reconstruction without degrading the resulting images. Starting with a mesh scale length of the order of the electrode separation it is shown that, for data obtained at presently achievable signal-to noise ratios of 60 to 80 dB, one or two refinement stages are sufficient to generate high quality images. PMID- 11876226 TI - The number of electrodes and basis functions in EIT image reconstruction. AB - In electrical impedance tomography, many factors affect the image reconstruction results. Among them are the number of electrodes (NOE) and the number of conductivity basis functions (NOCBF) for image reconstruction. The NOCBF generally reflects the density of the mesh with which images are reconstructed. How and to what extent do these factors affect the image reconstruction and corresponding images? In this area detailed analysis is still lacking. This study aims to address the above question. In this study, image reconstruction and its ill-posed condition were analysed by singular value decomposition (SVD) and spectral expansion theory with different configurations of NOE and NOCBF. The results in this study indicate that for a circular 2D plane object with electrodes evenly located around the boundary: (1) Under certain conditions, increasing the NOE enables us to improve the ill-posed condition in image reconstruction and hence improve the image quality. Generally more improvement is expected near the image periphery than in the image centre. (2) Increasing the NOCBF generally worsens the ill-posed condition. But it enables the solution to be sought in a finer subspace and may be able to improve the image quality on the periphery, while generally the result in image centre depends more on the prior information incorporated in the regularization. PMID- 11876227 TI - Analytical solution to the three-dimensional electrical forward problem for an elliptical cylinder. AB - In electrical impedance tomography, the shape of the object being imaged (such as the human thorax) is often complex. For this reason, numerical techniques, such as finite element method, are often used for solving the forward problem in 3D rather than analytical solutions which can only model simple geometrical shapes. However, an analytical solution to the 3D forward problem can often be useful. This paper will present an analytical solution to the forward problem for an elliptical cylinder whose eccentricity can be easily modified to approximate the shape of the human thorax. PMID- 11876228 TI - Design and performance of the UCLH mark 1b 64 channel electrical impedance tomography (EIT) system, optimized for imaging brain function. AB - The UCLH Mark 1b is a portable EIT system that can address up to 64 electrodes, which has been designed for imaging brain function with scalp electrodes. It employs a single impedance-measuring circuit and multiplexer so that electrode combinations may be addressed flexibly using software. It operates in the relatively low frequency band between 225 Hz and 77 kHz, as lower frequencies produce larger changes during brain activity, and has a videocassette-sized headbox on a lead 10 m long, connected to a base box the size of a video recorder, and notebook PC, so that recordings may be made in ambulant subjects. Its performance was assessed using a resistor-capacitor network, and two saline filled tanks-a cylindrical Perspex one and a latex one which contained a human skull. System signal-to-noise ratio was better than 50 dB and the maximum reciprocity error less than 10% for most frequencies. The CMMR was better than 80 dB at 38 kHz and a sponge, 20 mm across, which caused a local 12% impedance increase, was correctly localized in images. This suggests that the system has adequate performance to image impedance changes of 5-50% known to occur in the brain during normal activity, epilepsy or stroke; clinical trials to image these conditions are in progress. PMID- 11876229 TI - Modelled current distribution in cervical squamous tissue. AB - The electrical properties of cervical squamous epithelium have been modelled in the frequency range 100 Hz to 10 MHz. The hierarchical modelling process comprises a cellular level stage, which includes detailed models of cells typical of different depths within the epithelium and a tissue model, which utilizes electrical properties obtained from the cellular models. The fit between the modelled and measured impedance spectra and the distribution of current with depth depends on the macroscopic model structure. Both the properties of the basement membrane and the presence of a surface mucus layer are shown to have a significant effect. The best fit with measured data is obtained when a 10 microm thick, high-conductivity surface layer is included in the tissue model. PMID- 11876230 TI - A comparison of methods for measurement of spatial resolution in two-dimensional circular EIT images. AB - The literature concerning measurement of spatial resolution in electrical impedance tomography (EIT) is vague. Different groups often use their own method or a modified version of a better known method, thus hindering a generalized resolution measurement which could be useful for gauging the performance of one system against another. Measurement of spatial resolution in EIT is further complicated by its spatial variant nature and hence cannot be expressed simply with a single parameter as it can be in other imaging modalities (such as nuclear medicine or MRI for example). If the performance of each acquisition and image reconstruction system in EIT is to be compared objectively then there needs to be a common standard. In this paper the results of different methods for calculating spatial resolution are compared and an improved method is proposed which aims to fulfil this role. PMID- 11876231 TI - Indirect calculation of breast tissue impedance values. AB - One of the problems facing anyone attempting the investigation of dielectric properties of living tissue is the presence of skin, which screens all that lies under it from direct measurement. Thus, in non-invasive breast examination using transimpedance measurements, skin parameters heavily influence the results, specifically at low (less than 10 kHz) frequencies. In this paper a method for overcoming this difficulty by using multi-frequency measurements obtained from a surface current distribution over a flat probe is described. By using the variation in the shape of the real and imaginary parts of the surface current density at different frequencies, the original dielectric values of the skin and the underlying tissue can be obtained, based on the assumption of the existence of a two-layer geometry, with the upper (skin) layer much thinner than the lower (tissue) layer. The results obtained can be used in the diagnosis of breast cancer using existing transimpedance measurement devices. PMID- 11876232 TI - Resistivity changes in conductive silicone sheets under stretching. AB - This paper reports a preliminary finding associated with an investigation of how tissues respond to mechanical stress. The stress distribution within the tissue may be the result of normal function, for example, joint forces, or it may result from interventions such as tissue suturing during or after surgery. We sought to combine electrical and mechanical computational models in order to better understand the interaction between the two. For example, if mechanical stress is applied to tissue this may change the cell arrangements within the tissue matrix and hence change the electrical properties. If this interaction could be determined, then it should be possible to use electrical impedance tomography measurements to identify stress patterns in tissues. Measurements of resistivity changes have been made in conductive silicone rubber sheets when subject to a uniaxial stress of up to 10%. Relatively large changes in resistivity are produced (up to 200%). These changes are far larger than those predicted arising from topological changes alone. It is suggested that under stress the conductive islands of carbon within the silicone rubber sheet undergo a reversible disassociation from their neighbours and that the material's electrical properties change under load. If similar stress-resistivity relationships occur within biological materials it may be possible to recover the stress fields within tissues from transfer impedance measurements and thereby predict if actions such as inappropriate suture tension will compromise tissue viability. PMID- 11876233 TI - Frequency downconversion and phase noise in MIT. AB - High-frequency (3-30 MHz) operation of MIT systems offers advantages in terms of the larger induced signal amplitudes compared to systems operating in the low- or medium-frequency ranges. Signal distribution at HF, however, presents difficulties, in particular with isolation and phase stability. It is therefore valuable to translate received signals to a lower frequency range through heterodyne downconversion, a process in which relative signal amplitude and phase information is in theory retained. Measurement of signal amplitude and phase is also simplified at lower frequencies. The paper presents details of measurements on a direct phase measurement system utilizing heterodyne downconversion and compares the relative performance of three circuit configurations. The 100-sample average precision of a circuit suitable for use as a receiver within an MIT system was 0.008 degrees for input amplitude -21 dBV. As the input amplitude was reduced from -21 to -72 dBV variation in the measured phase offset was observed, with the offset varying by 1.8 degrees. The precision of the circuit deteriorated with decreasing input amplitude, but was found to provide a 100-sample average precision of <0.022 degrees down to an input amplitude of -60 dBV. The characteristics of phase noise within the system are discussed. PMID- 11876234 TI - Sensitivity maps for low-contrast perturbations within conducting background in magnetic induction tomography. AB - Magnetic induction tomography (MIT) is a contactless method for mapping the electrical conductivity of tissue by measuring the perturbation of an alternating magnetic field with appropriate receiver coils. Reconstruction algorithms so far suggested for biomedical applications are based on weighted backprojection, hence requiring tube-shaped zones of sensitivity between excitation coils and receiving coils, the sensitivity being essentially zero outside this 'projection beam'. This condition is met for conducting perturbations in empty space and for some special configurations of insulators in saline. In biological structures, however, perturbations with low conductivity contrast are embedded into a bulk conductor. The respective sensitivity distribution was investigated and quantified theoretically and experimentally by displacing a conducting (agar, 8 S m(-1)) and an insulating sphere within a saline tank (4 S m(-1)). In contrast to the case in the empty space the sensitivity is not confined to a tube but even increases outside the 'projection beam'. The difference can be explained by the interaction of bulk currents with the perturbing object. This effect invalidates backprojection and hence the solution of the complete inverse eddy-current problem is suggested. PMID- 11876235 TI - Phasic three-dimensional impedance imaging of cardiac activity. AB - Electrical impedance images were made using the ACT 3 instrument, which applies currents simultaneously to 32 electrodes and measures the resulting voltages on those same electrodes. A reconstruction algorithm was written for a three dimensional cylinder having electrodes in two or four layers, using current patterns that pass current among different planes of electrodes, as well as within each plane. We have previously reported useful vertical resolution by the use of added layers of electrodes. The aim of the present study was to demonstrate that physiologically useful information can be obtained by examining cephalo-caudal differences in three-dimensional images. Phasic changes throughout the cardiac cycle are seen to be markedly different at the heart compared to lung region, both above and beside it. We formed hydrogel electrodes each 3 cm tall and 7 cm wide and applied them to the thorax of an upright human subject in four horizontal rows; each row contained eight electrodes. During breath-holding, cardiac activity was seen in all layers. With systole, conductivity in the anterior of the lowest layers decreased, but not in the upper layer. In the upper layers, conductivity increased with systole in many regions. These observations are consistent with the opposite changes in blood volume of the heart and lungs and the locations of these organs. This paper demonstrates the feasibility of producing and displaying physiologically interpretable three-dimensional images of the chest in real time. PMID- 11876236 TI - Use of electrical impedance tomography (EIT) for the assessment of unilateral pulmonary function. AB - We describe a fully automatable quantification process for the assessment of unilateral pulmonary function (UPF) by means of EIT and propose a measurement protocol for its clinical implementation. Measurements were performed at the fourth and sixth intercostal levels on a first group of ten healthy subjects (5M, 5F, ages 26-48 years) to define the proper protocol by evaluating the most common postures and ventilation modes. Several off-line processing tools were also evaluated, including the use of digital filters to extract the respiratory components from EIT time series. Comparative measures were then carried out on a second group consisting of five preoperatory patients with lung cancer (4M, IF, ages 25-77 years) scheduled for radionuclide scanning. Results show that measurements were best performed with the subject sitting down, holding his arms up and breathing spontaneously. As regards data processing, it is best to extract Fourier respiratory components. The mean of the healthy subject group leads to a left-right division of lung ventilation consistent with literature values (47% left lung, 53% right lung). The comparative study indicates a good correlation (r = 0.96) between the two techniques, with a mean difference of (-0.4+/-5.4)%, suggesting that the elimination of cardiac components from the thoracic transimpedance signal leads to a better estimation of UPF. PMID- 11876237 TI - Imaging the breast with EIS: an initial study of exam consistency. AB - Use of electrical impedance spectroscopy (EIS) to image the breasts of women with both normal and abnormal conditions requires the ability to deliver a consistent and repeatable exam. To investigate the degree to which our current imaging system can meet this requirement we conducted an initial study of exam consistency. The trial involved the imaging of 25 breasts stratified into four separate substudies with increasing levels of electrode placement uncertainty. The degree of complexity ranged from single-placement single-session imaging to multiple-placement single-session imaging to multiple-placement multiple-session imaging. Both visual analysis and quantitative comparisons using mean squared difference (MSD) measures between pairs of permittivity and conductivity images were performed. A new breast interface with the improved vertical and radial electrode array positioning capability required to complete this study is described. Not surprisingly, the results show a dominant trend of increased image variability with increased electrode placement uncertainty. Importantly, quantitative levels of image consistency are reported through MSD analysis. On average across all frequencies analysed, MSDs for single placements are well below 1%, near 2-3% for repositioned breasts during the same session and approximately 15% for re-examined breasts in multiple sessions conducted over time. Overall, these results suggest that EIS breast exams are consistent provided the electrode placement is well controlled, typically with better than 1 cm accuracy. PMID- 11876238 TI - A maximum entropy method to compute the 13NH3 pulmonary transit time from right to left ventricle in cardiac PET studies. AB - The distribution function of pulmonary transit times (fPTTs) contains information on the transit time of blood through the lungs and the dispersion in transit times. Most of the previous studies have used specific functional forms with adjustable parameters to characterize the fPTT. It is the purpose here to investigate the possibility of estimating the fPTT in a model-free way. The method employs the maximum entropy principle and is used, in particular, on cardiac positron emission tomographic (PET) studies but is believed to be more generally applicable. Using this principle in a test case, we were able to accurately identify a two-peaked transfer function, which may theoretically be seen in patients with pulmonary disease confined to one lung. Transit time values for [13N]-ammonia were produced by applying the algorithm to PET studies from normal volunteers. PMID- 11876239 TI - Editorial: Biomedical applications of electrical impedance tomography. PMID- 11876240 TI - Preliminary static EIT images of the thorax in health and disease. AB - The results of a preliminary clinical evaluation of a one-frequency electrical impedance tomography (EIT) system enabling static in vivo imaging are presented. The design of the measuring system and image reconstruction software are described. Thirty-one subjects were examined and divided into four clinical groups. The first group consisted of 22 patients with clinical diagnosis of lung cancer with tumour localization in one lung. The second group consisted of seven healthy subjects. A patient after a one-sided pneumectomy and another with one sided emphysema diagnosis were also examined. Static EIT images of a healthy human chest and a chest with various abnormalities are given and discussed. The evaluated system distinguishably visualizes various states of lungs and thorax including lung cancer. The average static conductivity of an affected lung in the first clinical group statistically differs from the average conductivity of a healthy lung. In spite of low spatial resolution, according to preliminary results, the method can be sensitive to cancer and other lung diseases in screening investigations. PMID- 11876241 TI - Comparison of segmental and global bioimpedance spectroscopy errors using generalizability theory. AB - The generalizability theory, an expansion of classic true-score reliability theory, was used to investigate the generalizability of observed segmental extracellular fluid (ECF) and intracellular fluid (ICF) distribution measurements. The test instrument was a Xitron Hydra ECF/ICF bioimpedance analysis system model 4200, Xitron Technologies, San Diego, CA. Fifty American healthy men (17-72 years) and 50 American healthy women (17-76 years) volunteered as participants. Xitron continuous segmental ECF-ICF procedures for testing leg segmental data were followed for testing participants in both the standing erect and lying supine postures. A two-facet, person-by-trial, completely crossed design was used. All facets were treated as random. During a one-day session each subject was tested involving 20 trials for the standing erect posture and 20 trials for the lying supine posture. Data on each fluid measurement, each body posture and each sex group were independently analysed. The analyses revealed that the trial factor accounted for less than 0.2% of the total variance for ECF and ICF scores. ECF and ICF generalizability coefficients for the segmental method were 0.99 or greater. In comparing ECF segmental to ECF global, results showed generalizability coefficients were similar. However, ICF segmental coefficients were larger than the coefficients produced by the global method. In conclusion, the segmental method appeared more reliable than the global method, under the conditions of this study. PMID- 11876242 TI - Assessment of dynamic cerebral autoregulation based on spontaneous fluctuations in arterial blood pressure and intracranial pressure. AB - Assessments of dynamic cerebral autoregulation usually measure the cerebral blood flow velocity (CBFV) response to changes in arterial blood pressure (ABP). We studied the effect of substituting ABP by cerebral perfusion pressure (CPP), expressed as the difference between ABP and intracranial pressure (ICP), in estimates of dynamic autoregulation obtained by transfer function analysis. CBFV, ABP and ICP were recorded during periods of physiological stability in 30 patients with severe head injury. Transfer function analysis was performed using the following combinations of input-output variables: ABP-CBFV, CPP-CBFV and CBFV ICP. Frequency and time-domain (step response) functions were averaged for recordings with mean ICP < 20 mmHg (group A) and mean ICP > or = 20 mmHg (group B). The ABP-CBFV transfer function parameters and step response for group A were similar to previous studies in normal subjects, but group B showed deterioration of dynamic autoregulation. Radically different step responses were obtained from both groups for the CPP-CBFV transfer function and the coherence was not significantly improved. The CBFV-ICP transfer function had the highest values of coherence and indicates that changes in CBFV are the cause of spontaneous fluctuations in ICP. Furthermore, the ICP step response plateau was significantly higher for group B than for group A. An alternative calculation of the CBFV step response to changes in CPP resembled the corresponding responses for the ABP input. For spontaneous fluctuations in ABP, ICP and CBFV, it is not possible to calculate the CPP-CBFV transfer function directly due to the high positive correlation between ICP and CBFV, but an alternative estimate can be obtained by using the CBFV-ICP transfer function. The latter could also be useful as a method to assess intracranial compliance in head injury patients. PMID- 11876243 TI - The repeatability of cerebral autoregulation assessment using sinusoidal lower body negative pressure. AB - A forced periodic variation in blood pressure produces a similar variation in cerebral blood velocity. The amplitudes and phases of the pressure and velocity waveforms are indicative of the dynamic response of the cerebral autoregulation. The phase of the velocity leads the pressure; the greater the phase difference the faster the autoregulation response. Various techniques have been employed to oscillate arterial blood pressure but measurement reproducibility has been poor. The purpose of this study was to assess the reproducibility of phase measurements when sinusoidal lower body negative pressure is used to vary blood pressure. Five healthy volunteers were assessed at two vacuum levels on each of eight visits. For each measurement a 12 s sinusoidal cycle was maintained for 5 min. The Fourier components of blood pressure and the middle cerebral artery velocity were determined at the oscillation frequency. The phase of velocity consistently led the pressure. The mean phase difference was 42+/-13 degrees for the stronger vacuum and 36+/-42 degrees for the weaker vacuum. The variation given is the within-subjects standard deviation estimated from a one-way analysis of variance. Sinusoidal lower body negative pressure is a useful stimulus for investigating autoregulation; it has advantages over other methods. High vacuums show good reproducibility but are too uncomfortable for patient use. PMID- 11876244 TI - The difference in pulse transit time to the toe and finger measured by photoplethysmography. AB - Blood pressure pulse wave velocity (PWV) is a parameter which is related to arterial distensibility. Its direct assessment, by measuring the appearance time of a pressure pulse in two sites along an artery and the distance between the two sites, is complicated and inaccurate. In the current study, pulse transit time (PTT) to the toes and fingers of 44 normotensive male subjects was measured by photoplethysmography (PPG) and ECG. The arrival time of the pulses at the toe and finger was determined from the foot of the systolic rise of the PPG signal, i.e. at end-diastolic time. Two parameters, which are related to PWV, were tested: the time delay between the ECG R-wave and the arrival time of the pulses at the toe (E-T PTT), and the difference in the transit time of the blood pressure pulses between the toe and finger (T-F PTTD). E-T PTT and T-F PTTD decreased as functions of the subject's age and systolic blood pressure (SBP), but their dependence on the diastolic blood pressure (DBP) was not statistically significant. The decrease of the PTT parameters with age is attributed to the direct structural decrease of the arterial compliance with age and not to functional effects associated with the increase of the blood pressure with age, since the PTT parameters did not depend on DBP though the measurements were performed at end-diastole. PMID- 11876245 TI - Comparison of algorithms for non-linear inverse 3D electrical tomography reconstruction. AB - Non-linear electrical impedance tomography reconstruction algorithms usually employ the Newton-Raphson iteration scheme to image the conductivity distribution inside the body. For complex 3D problems, the application of this method is not feasible any more due to the large matrices involved and their high storage requirements. In this paper we demonstrate the suitability of an alternative conjugate gradient reconstruction algorithm for 3D tomographic imaging incorporating adaptive mesh refinement and requiring less storage space than the Newton-Raphson scheme. We compare the reconstruction efficiency of both algorithms for a simple 3D head model. The results show that an increase in speed of about 30% is achievable with the conjugate gradient-based method without loss of accuracy. PMID- 11876246 TI - Fractal characterization of complexity in temporal physiological signals. AB - This review first gives an overview on the concept of fractal geometry with definitions and explanations of the most fundamental properties of fractal structures and processes like self-similarity, power law scaling relationship, scale invariance, scaling range and fractal dimensions. Having laid down the grounds of the basics in terminology and mathematical formalism, the authors systematically introduce the concept and methods of monofractal time series analysis. They argue that fractal time series analysis cannot be done in a conscious, reliable manner without having a model capable of capturing the essential features of physiological signals with regard to their fractal analysis. They advocate the use of a simple, yet adequate, dichotomous model of fractional Gaussian noise (fGn) and fractional Brownian motion (fBm). They demonstrate the importance of incorporating a step of signal classification according to the fGn/fBm model prior to fractal analysis by showing that missing out on signal class can result in completely meaningless fractal estimates. Limitation and precision of various fractal tools are thoroughly described and discussed using results of numerical experiments on ideal monofractal signals. Steps of a reliable fractal analysis are explained. Finally, the main applications of fractal time series analysis in biomedical research are reviewed and critically evaluated. PMID- 11876247 TI - Portal vein thrombosis and pancreatic failure. AB - We report a case of a child with portal vein thrombosis presenting with protein losing enteropathy. He later developed exocrine and endocrine pancreatic failure. This association has not been reported before. PMID- 11876248 TI - Pancreatic arteriovenous malformation associated with duodenal ulcer and H. pylori infection. AB - Pancreatic arteriovenous malformation (PAVM) is a rare condition that may cause duodenal ulcer. A 36-yr-old man with PAVM associated with duodenal ulcer and H. Pylori infection is described. The patient had recurrent episode of upper abdominal pain despite healed ulcer and H. Pylori eradication. The preoperative diagnosis was confirmed by computed tomography and the patient was treated with a pancreatoduodenectomy. Histological examination of the resected pancreas revealed a pancreatic arteriovenous malformation involving the adjacent duodenal wall. PMID- 11876249 TI - Small-bowel obstruction secondary to subcutaneous small-bowel entrapment: a late complication of laparostomy for necrotizing pancreatitis. AB - Laparostomy is a well recognized strategy for the management of patients who have necrotizing pancreatitis and may require multiple re-intervention. The open wound can be left to heal through a process of granulation and contraction. This article describes intestinal obstruction secondary to entrapment of a loop of small bowel within the cicatrix of the contracting cutaneous scar. An awareness of the potential for entrapment of the small bowel in the healing scar is critical for clinicians using laparostomy in the management of acute necrotizing pancreatitis. PMID- 11876250 TI - The latest in pancreatic cancer research: Lustgarten Foundation awardees present their findings. PMID- 11876251 TI - In vivo effect of pancreatic phospholipase A2 on the arachidonic acid cascade. AB - BACKGROUND: In acute pancreatitis, pancreatic phospholipase A2 increases in systemic circulation. Yet the pathophysiological significance is controversial, because previous in vitro studies have shown that the enzyme has little cytotoxicity or ability to activate the arachidonic acid cascade by itself in contrast to other isozymes. AIM OF THE STUDY: The aims of this study are to examine the effect of pancreatic phospholipase A2 on the arachidonic acid cascade in vivo; to explain the discrepancy, if present, between in vitro and in vivo findings; and to reassess the pathophysiological significance of circulating pancreatic phospholipase A2. METHODS: Pancreatic phospholipase A2 was infused intravenously in guinea pigs, and changes in the arachidonic acid cascade, plasma lipoprotein, and cardiopulmonary function were investigated. RESULTS: Plasma concentrations of 6-keto-prostaglandin F1alpha, prostaglandin E2, and thromboxane B2 increased after intravenous (iv) infusion of pancreatic phospholipase A2. Some of the plasma phospholipids such as phosphatidylcholine and phosphatidylethanolamine decreased, and free dihomo-gamma-linolenic acid, arachidonic acid, and eicosapentaenoic acid were detected in plasma. These changes were accompanied with decreases in blood pressure, heart rate, and base excess. CONCLUSION: Circulating pancreatic phospholipase A2 activates the arachidonic acid cascade, probably by supplying free eicosanoid precursors from plasma lipoprotein to eicosanoid-producing cells. It is supposed to be a cause of systemic complications in acute pancreatitis. PMID- 11876252 TI - Distribution of nitric oxide synthase and secretory role of exogenous nitric oxide in the isolated rat pancreas. AB - BACKGROUND: Pancreatic production and in vivo effects of nitric oxide (NO) have been shown by several studies. In order to examine the direct actions of the NO donor sodium nitroprusside (SNP), this study used in vitro specimens of the rat pancreas where the distribution of neuronal nitric oxide synthase (NOS) and the secretory effects of SNP and the cyclic GMP (cGMP) analog 8-bromo cyclic GMP (8 Br cGMP) were investigated. METHODS: NO containing pancreatic nerves were visualized by NOS immunohistochemistry. Basal and stimulated amylase output from rat pancreatic segments was measured by an on-line fluorimetric method. Stimulation was achieved by either acetylcholine (ACh) or electrical field stimulation (EFS). Intracellular free calcium concentration ([Ca2+]i) was measured in dispersed pancreatic acinar cells. RESULTS: NOS containing nerves were demonstrated in the vicinity of pancreatic acini and blood vessels. SNP and 8-Br cGMP inhibited both basal and EFS evoked amylase output but failed to inhibit ACh induced amylase output. Basal [Ca2+]i was decreased by both SNP and 8 Br cGMP but neither SNP nor 8-Br cGMP influenced the ACh evoked increase in [Ca2+]i. CONCLUSION: NO is well distributed in the rat exocrine pancreas. Exogenous nitric oxide may have a dual action in the isolated rat pancreas: Inhibition of basal amylase secretion in acinar cells and inhibition of ACh release from intrinsic nerve terminals. Both effects seem to be calcium dependent and possibly mediated by cGMP. PMID- 11876253 TI - Expression of the IIIc variant of FGF receptor-1 confers mitogenic responsiveness to heparin and FGF-5 in TAKA-1 pancreatic ductal cells. AB - BACKGROUND: Fibroblast growth factors (FGFs) contribute to angiogenesis and mitogenesis by binding to tyrosine kinase receptors termed FGF receptors (FGFRs). FGF-5 is a secreted FGF that is believed to preferentially act via the IIIc splice variant of FGFR-1. Human pancreatic ductal carcinoma cells express FGF-5 and FGFR-1IIIc, implying a potential for autocrine growth modulation. AIM: In this study we investigated the importance of FGFR-1 IIIc expression for FGF-5 mitogenic signaling in a pancreatic ductal cell line. METHODS: A cDNA encoding FGFR-1 IIIc was expressed in the well-differentiated TAKA-1 Syrian hamster pancreatic ductal cell line. RESULTS: TAKA-1 cells secrete FGF-5, but were found not to express FGFR-1 and to be unresponsive to exogenous FGF-5. In contrast, TAKA-1 clones expressing FGFR-1 IIIc were growth stimulated in the presence of FGF-5 and displayed enhanced mitogen-activated protein kinase (MAPK) activity in the presence of FGF-5. PD98059, an inhibitor of this pathway, inhibited FGF-5 induced growth in these clones. CONCLUSION: Our data demonstrate that FGFR-1 IIIc can mediate FGF-5-induced mitogenesis via the MAPK pathway in pancreatic ductal cells, and suggest that expression of FGFR-1 IIIc in conjunction with FGF-5 may contribute to the pathobiology of human pancreatic cancer. PMID- 11876254 TI - Neurofibromatosis-1 and ampullary gangliocytic paraganglioma causing biliary and pancreatic obstruction. AB - Gangliocytic paragangliomas (GPs) are exceedingly rare tumors that arise in close proximity of the papilla of Vater. Nevertheless, jaundice is an uncommon presenting symptom, reported in only 3 of 125 cases described in the literature to date, with gastrointestinal bleeding being more common. Association between GPs and neurofibromatosis 1 (NF-1), described in two patients, may be more than casual, being the frequency in normal population 1:3,000. We report an additional case of this association, presenting for the first time with simultaneous obstruction of the common biliary duct, and of the main pancreatic duct. Previous cases of GPs with jaundice and/or NF-1 are reviewed, and updating of the histogenesis presented. PMID- 11876255 TI - Cavernous pancreatic ductal ectasia with smooth muscle proliferation causing recurrent acute pancreatitis. AB - BACKGROUND: Pancreatic cystic lesions have various etiologies, including pseudocyst (inflammatory cyst), retention cyst, congenital cyst, and neoplastic cyst. RESULTS: This report describes a previously unreported, unique pancreatic cyst-like lesion causing recurrent acute pancreatitis. A 23-yr-old man had an 8 x 5 x 3-cm pancreatic head mass which contained multiple 3-7-mm cysts communicating with the main pancreatic duct on imaging studies. Pancreatoduodenectomy with mass excision prevented further attacks of acute pancreatitis. Pathological examination showed multiple cystic dilatations of branch pancreatic ducts surrounded by proliferating smooth muscle tissue, probably associated with hamartomatous changes. CONCLUSION: We consider the present lesion to represent cavernous pancreatic ductal ectasia with smooth muscle proliferation because of its striking cholangiopancreatographic similarity to Caroli disease. PMID- 11876257 TI - Potential bile acid metabolites. 24. An efficient synthesis of carboxyl-linked glucosides and their chemical properties. AB - A facile and efficient synthesis of the carboxyl-linked glucosides of bile acids is described. Direct esterification of unprotected bile acids with 2,3,4,6-tetra O-benzyl-D-glucopyranose in pyridine in the presence of 2-chloro-1,3,5 trinitrobenzene as a coupling agent afforded a mixture of the alpha- and beta anomers (ca. 1:3) of the 1-O-acyl-D-glucoside benzyl ethers of bile acids, which was separated effectively on a C18 reversed-phase chromatography column (isolated yields of alpha- and beta-anomers are 4-9% and 12-19%, respectively). Subsequent hydrogenolysis of the alpha- and beta-acyl glucoside benzyl ethers on a 10% Pd-C catalyst in acetic acid/methanol/EtOAc (1:2:2, by vol) at 35 degrees C under atmospheric pressure gave the corresponding free esters in good yields (79-89%). Chemical specificities such as facile hydrolysis and transesterification of the acyl glucosides in various solvents were also discussed. PMID- 11876258 TI - Dibutyrate derivatization of monoacylglycerols for the resolution of regioisomers of oleic, petroselinic, and cis-vaccenic acids. AB - Dibutyrate derivatives of monoacylglycerols of oleic, petroselinic, and cis vaccenic acids were prepared by diesterification of monoacylglycerols with n butyryl chloride. The resulting triacylglycerols were analyzed by gas chromatography (GC) with a 65% phenyl methyl silicone capillary column and separated on the basis of both fatty acid composition and regiospecific position. The petroselinic acid derivatives eluted first, followed sequentially by the oleic and cis-vaccenic acid derivatives, with the sn-2 positional isomer eluting before the sn-1 (3) isomer in each case. Separation of the peaks was almost baseline between petroselinic and oleic acids as well as between oleic and cis vaccenic acids. To assess the accuracy of the method, mixtures of triolein, tripetroselinin, and tri-cis-vaccenin in various known proportions were partially deacylated with the use of ethyl magnesium bromide and derivatized and analyzed as above. The results showed that this method compares favorably to the existing methods for analysis of oleic, petroselinic, and cis-vaccenic fatty acids by GC with respect to peak separation and accuracy, and it also provides information on the regiospecific distribution of the fatty acids. The method was applied to basil (Ocimum basilicum) and coriander (Coriandrum sativum) seed oils. cis Vaccenic, oleic, and linoleic acids were mainly distributed at the sn-2 position in basil seed oil, and higher proportions of linolenic, palmitic, and stearic acids were distributed at the sn-1(3) position than at the sn-2 position. In coriander seed oil, petroselinic acid was mainly distributed at the sn-1 (3) position, and both oleic and linoleic acids were mostly located at the sn-2 position, whereas palmitic, stearic, and cis-vaccenic acids were located only at the sn-1 (3) position. PMID- 11876259 TI - Abietoid seed fatty acid compositions--a review of the genera Abies, Cedrus, Hesperopeuce, Keteleeria, Pseudolarix, and Tsuga and preliminary inferences on the taxonomy of Pinaceae. AB - The seed fatty acid (FA) compositions of Abietoids (Abies, Cedrus, Hesperopeuce, Keteleeria, Pseudolarix, and Tsuga) are reviewed in the present study in conclusion to our survey of Pinaceae seed FA compositions. Many unpublished data are given. Abietoids and Pinoids (Pinus, Larix, Picea, and Pseudotsuga) constituting the family Pinaceae-are united by the presence of several delta5 olefinic acids, taxoleic (5,9-18:2), pinolenic (5,9,12-18:3), coniferonic (5,9,12,15-1 8:4), keteleeronic (5,11-20:2), and sciadonic (5,11,14-20:3) acids, and of 14-methyl hexadecanoic (anteiso-17:0) acid. These acids seldom occur in angiosperm seeds. The proportions of individual delta5-olefinic acids, however, differ between Pinoids and Abietoids. In the first group, pinolenic acid is much greater than taxoleic acid, whereas in the second group, pinolenic acid is greater than or equal to taxoleic acid. Moreover, taxoleic acid in Abietoids is much greater than taxoleic acid in Pinoids, an apparent limit between the two subfamilies being about 4.5% of that acid relative to total FA. Tsuga spp. appear to be a major exception, as their seed FA compositions are much like those of species from the Pinoid group. In this respect, Hesperopeuce mertensiana, also known as Tsuga mertensiana, has little in common with Abietoids and fits the general FA pattern of Pinoids well. Tsuga spp. and H. mertensiana, from their seed FA compositions, should perhaps be separated from the Abietoid group and their taxonomic position revised. It is suggested that a "Tsugoid" subfamily be created, with seed FA in compliance with the Pinoid pattern and other botanical and immunological criteria of the Abietoid type. All Pinaceae genera, with the exception of Pinus, are quite homogeneous when considering their overall seed FA compositions, including delta5-olefinic acids. In all cases but one (Pinus), variations from one species to another inside a given genus are of small amplitude. Pinus spp., on the other hand, have highly variable levels of delta5 olefinic acids in their FA compositions, particularly when sections (e.g., Cembroides vs. Pinus sections) or subsections (e.g., Flexiles and Cembrae subsections from the section Strobus) are compared, although they show qualitatively the same FA patterns characteristic of Pinoids. Multicomponent analysis of Abietoid seed FA allowed grouping of individual species into genera that coincide with the same genera otherwise characterized by more classical botanical criteria. Our studies exemplify how seed FA compositions, particularly owing to the presence of delta5-olefinic acids, may be useful in sustaining and adding some precision to existing taxonomy of the major family of gymnosperms, Pinaceae. PMID- 11876260 TI - Assessment of dietary and genetic factors influencing serum and adipose fatty acid composition in obese female identical twins. AB - Fourteen pairs of obese female monozygotic twins were recruited for a study of genetic influences on serum and adipose fatty acid (FA) composition. Following 1 wk of inpatient stabilization, fasting serum and adipose tissue obtained by surgical excision were analyzed by thin-layer and gas chromatography. Intrapair resemblances (IPR) for individual FA were assessed by Spearman rank correlation and by analysis of variance and were found in serum cholesteryl esters (CF), triglycerides (TG), and adipose TG. With two exceptions (CE linoleate and adipose eicosapentaenoate), these IPR were limited to the nonessential FA. Palmitate had significant IPR in four lipid fractions; in serum CE and adipose TG palmitate was strongly correlated with multiple measures of adiposity. In contrast to other lipid fractions, serum phosphatidylcholine (PC) FA had 12 [PR, of which 6 were essential FA including arachidonate (r = 0.76, P < 0.0005), eicosapentaenoate (r = 0.78, P < 0.0005), and docosahexaenoate (r = 0.86, P< 0.0001). The PC [PR could not be explained by analysis of preadmission 7-d food records. After dividing the pairs into two groups differing and nondiffering according to fat intake of individuals in the pair, there was no evidence of a gene-environment interaction between fat intake and FA composition. The IPR for nonessential FA indicate that there is active genetic control of either food choices or postabsorptive metabolic processing. The high level of IPR in the PC fraction in contrast to the other lipid fractions suggests strong genetic influence over selection of specific FA for this membrane fraction independent of diet. PMID- 11876261 TI - Plant sterol esters lower plasma lipids and most carotenoids in mildly hypercholesterolemic adults. AB - The ability of plant sterol esters (PSE) in salad dressing to modify plasma lipids and carotenoids was determined in 26 men and 27 women fed controlled, weight-maintaining, isocaloric diets. Diets contained typical American foods that provided 32% of energy from fat. Dressings contained 8 g (ranch) or 4 g (Italian) of fat per serving. PSE (3.6 g/d) were provided in two servings/d of one of the dressings. Diets with ranch or Italian dressing without and with PSE were fed for 3 wk/diet and crossed over randomly within dressings. Diets were adjusted to similar fat and fatty acid concentrations. Type of salad dressing did not affect plasma lipids, lipoproteins, carotenoids, or fat-soluble vitamins (P > 0.05). Switching from a self-selected baseline diet to the control diet resulted in reduction in low density lipoprotein (LDL) cholesterol of 7.9%, a decrease in high density lipoprotein (HDL) cholesterol of 3.1%, and a decrease in triglycerides (TG) of 9.3%. Consumption of 3.6 g of PSE resulted in further decreases in LDL cholesterol (9.7%) and TG (7.3%) but no additional change in HDL cholesterol. Total plasma carotenoids decreased 9.6% with PSE. An automated stepwise procedure was developed to produce candidate mixed models relating plasma carotenoid response to PSE. These models adjusted for preintervention plasma carotenoid levels and effects of diets on blood lipids. There were significant decreases in beta-carotene, alpha-carotene, and beta-cryptoxanthin (females only) not associated with changes in plasma lipids. Plasma carotenoids on all diets remained within normal ranges. We conclude that low-fat foods, such as salad dressings, are effective carriers for PSE. PMID- 11876256 TI - Lipids and endothelium-dependent vasodilation--a review. AB - Studies using both in vitro and in vivo techniques have repeatedly shown that endothelium-dependent vasodilation (EDV) is impaired in different forms of experimental as well as human hypercholesterolemia. Clearly this impaired EDV can be reversed by lowering cholesterol levels by diet or medical therapy. Competitive blocking of L-arginine, changes in nitric oxide synthase activity, increased release of endothelin-1, and inactivation of nitric oxide due to superoxide ions all contribute to the impairment in EDV during dyslipidemia. The oxidation of low density lipoprotein, with its compound lysophosphatidylcholine, plays a critical role in these events. However, data on the role of triglycerides and fat-rich meals regarding EDV are not so consistent as data for cholesterol, although a view that the compositions of individual fatty acids and antioxidants are of major importance is emerging. Thus, this review shows that while impaired EDV is a general feature of hypercholesterolemia, the mechanisms involved and the therapeutic opportunities available still have to be investigated. Furthermore, discrepancies regarding the role of triglycerides and fat content in food may be explained by divergent effects of different fatty acids on the endothelium. PMID- 11876263 TI - Selective effect of cholesterylphosphoserine on intracellular cholesterol transport. AB - Cholesteryl-3beta-phosphoserine (CPHS) is a synthetic steroid affecting intracellular cholesterol transport. To compare CPHS with the well-known inhibitors progesterone and U18666A, we examined cholesterol transport in three human cell lines: the monocytic U-937, the endothelial ECV-304, and the lymphoid Jurkat. Under low density lipoprotein (LDL) loading, CPHS inhibited cholesterol esterification in U-937 and ECV-304 cells but not in Jurkat cells. In contrast, CPHS inhibited the mobilization of plasma membrane cholesterol induced by 25 hydroxycholesterol, brefeldin A, or sphingomyelinase in all cell lines. In cells pulse-labeled with [3 H]cholesterol, CPHS decreased incorporation of cholesterol and inhibited its esterification. In prelabeled cells, CPHS promoted cholesterol efflux and enhanced the cyclodextrin-mediated removal of plasma membrane cholesterol. CPHS did not affect endogenous cholesterol synthesis nor acylcoenzyme A:cholesterol acyltransferase activity. These data suggest that, unlike progesterone and U18666A, CPHS inhibits intracellular cholesterol transport by specifically affecting the movements of cholesterol in the plasma membrane. Owing to this restricted site of action, CPHS may help to clarify the role of the plasma membrane in cholesterol trafficking. For example, the lack of an effect of CPHS on the esterification of LDL-derived cholesterol in Jurkat cells suggests that most of the LDL-derived cholesterol in these cells is directly delivered to the endoplasmic reticulum without cycling through the plasma membrane. PMID- 11876262 TI - Exercise training-induced changes in sensitivity to endothelin-1 and aortic and cerebellum lipid profile in rats. AB - The purpose of this work was to study whether exercise training induces changes in the lipid profile of rat aorta and nervous system and in the in vitro intrinsic responsiveness of these tissues to endothel in-1 (ET-1) treatment. The exercise program performed successfully produced the characteristic metabolic alterations of the trained state. Exercise training induced a large and significant increase in the levels of both aortic ethanolamine plasmalogens (PlasEtn) and glucosylceramides. In contrast, a decrease of aortic ceramide and cholesterol levels was evoked by exercise training. ET-1 increased PlasEtn content only in sedentary animals. An exercise-induced increase in cerebellum levels of ceramides and ceramide monohexosides was found. The cerebellum ceramide content was increased by ET-1 more noticeably in sedentary rats than in trained animals. In contrast, cerebral cortex was observed to be largely insensitive to both exercise training and ET-1 treatment. It was concluded that exercise training (i) induces changes in both vascular and cerebellar lipid profiles, the former being much more pronounced than the latter, and (ii) diminishes the aortic and cerebellar sensitivity to ET-1 action. PMID- 11876264 TI - Reduced secretion of triacylglycerol in CaCo-2 cells transfected with intestinal fatty acid-binding protein. AB - The fatty acid-binding proteins are hypothesized to be involved in cellular fatty acid transport and trafficking. We established CaCo-2 cells stably transfected with intestinal fatty acid-binding protein (I-FABP) and examined how the expression of this protein may influence fatty acid metabolism. I-FABP expression was detectable in I-FABP-transfected cells, whereas parent CaCo-2 cells as well as mock-transfected cells failed to express detectable levels of I-FABP mRNA or protein at any stage of differentiation. For studies of lipid metabolism, cells were incubated with [14C]oleic acid in taurocholate micelles containing monoolein, and distribution of labeled fatty acid in cellular and secreted lipids was examined. In one transfected cell clone, expressing the highest level of I FABP, labeled cellular triacylglycerol increased approximately twofold as compared to control cells. The level of intracellular triacylglycerol in two other I-FABP-transfected clones resembled that of control cells. However, secretion of triacylglycerol was markedly reduced in all the I-FABP-expressing cell lines. Our data suggest that increased expression of I-FABP leads to reduced triacylglycerol secretion in intestinal cells. PMID- 11876265 TI - Regulation of phosphatidic acid phosphohydrolase 1 by fatty acids. AB - In the starved state and during metabolic stress, free fatty acids (FFA) are the principal hepatic energy supply, undergoing beta-oxidation. Accordingly, it appears paradoxical that FFA have been reported to increase the liver's esterification capacity by translocating the rate-limiting enzyme phosphatidic acid phosphohydrolase (PAP-1) from the cytosol to the endoplasmic reticulum. We have therefore investigated the regulation of rat liver PAP-1. Oleic acid inhibited PAP activity in all subcellular fractions, with PAP-1 activity in cytosol being the most sensitive. Inhibition was also observed with oleoyl-CoA, linoleate, and palmitate. Fatty acids and their derivatives show detergent effects at high concentrations, and such effects can lead to enzyme inhibition. Inhibition by oleate, however, was reversed by phosphatidic acid and albumin and exhibited sigmoidal kinetics. These results demonstrate that PAP-1 is reversibly inhibited by FFA and their CoA esters, which may play a role in directing hepatic FFA to beta-oxidation during times of increased energy demand. PMID- 11876266 TI - Formation of triacylglycerol core aldehydes during rapid oxidation of corn and sunflower oils with tert-butyl hydroperoxide/Fe2+. AB - The lipid ester core aldehydes formed during a rapid oxidation (7.8 M tert-butyl hydroperoxide, 90 min at 37 degrees C) of the triacylglycerols of purified corn and sunflower oils were isolated as dinitrophenylhydrazones by preparative thin layer chromatography and identified by reversed-phase high-performance liquid chromatography with on-line electrospray ionization mass spectrometry and by reference to standards. A total of 113 species of triacylglycerol core aldehydes were specifically identified, accounting for 32-53% of the 2,4 dinitrophenylhydrazine (DNPH)-reactive material of high molecular weight representing 25-33% of the total oxidation products. The major core aldehyde species (50-60% of total triacylglycerol core aldehydes) were the mono(9 oxo)nonanoyl- and mono(12-oxo)-9,10-epoxy dodecenoyl- or (12-oxo)-9-hydroxy-10,11 dodecenoyl-diacylglycerols. A significant proportion of the total (9-oxo)nonanoyl and epoxidized (12-oxo)-9,10-dodecenoyl core aldehydes was found in complex combinations with hydroperoxy or hydroxy fatty acyl groups (6-10% of total triacylglycerol core aldehydes). Identified were also di(9 oxo)nonanoylmonoacylglycerols (0.5% of total) and tri(9-oxo)nonanoylglycerols (trace). The identification of the oxoacylglycerols was consistent with the products anticipated from tert-butyl hydroperoxide oxidation of the major species of corn and sunflower oil triacylglycerols (mainly linoleoyl esters). However, the anticipated (13-oxo)-9,11-tridecadienoyl aldehyde-containing acylglycerols were absent because of further oxidation of the dienoic core aldehyde. A significant proportion of the unsaturated triacylglycerol core aldehydes contained tert-butyl groups linked to the unsaturated fatty chains via peroxide bridges (2-9%). The study demonstrates that rapid peroxidation with tert-butyl hydroperoxide constitutes an effective method for enriching natural oils and fats in triacylglycerol core aldehydes for biochemical and metabolic testing. PMID- 11876267 TI - Get your shoes on...! A response from a practicing dentist. PMID- 11876268 TI - A national oral health plan: implications and opportunities for research and academia. PMID- 11876269 TI - Overbite and overjet are not related to self-report of temporomandibular disorder symptoms. AB - Overbite and overjet, especially high or low values, have been found in some studies to be associated with temporomandibular disorders (TMD). This study evaluates the relationship between overbite/overjet and three TMD self-report measures (pain, joint noises, limited mouth-opening). Subjects were from two population-based cross-sectional studies (3033 subjects). After adjustment for age and gender, high or low values of overbite were not associated with an increased risk of self-reported TMD pain as compared with a reference category of a normal overbite of 2 to 3 mm (-8 to -1 mm, odds ratio = 0.36, 95% confidence interval = 0.05-2.76; 6 to 15 mm, odds ratio = 1.08, 95% confidence interval = 0.68-1.72). Similar non-significant results were found for overjet and TMD pain, and for the association of overjet/overbite and joint noises or limited mouth opening. This study provides the strongest evidence to date that there is no association between overbite or overjet and self-reported TMD. PMID- 11876270 TI - Cathepsin C, matrix metalloproteinases, and their tissue inhibitors in gingiva and gingival crevicular fluid from periodontitis-affected patients. AB - Successive active phases observed in periodontal diseases may be explained either by a sudden activation of the pro-forms of tissue-stored degradative enzymes such as metalloproteinases (MMPs) or by an imbalance between metalloproteinases and their tissue inhibitors (TIMPs). To discriminate between these two hypotheses, we quantified the levels, the percentage of active form, and the activities of four metalloproteinases (MMPs -1, -2, -3, and -9), as well as the levels of two tissue inhibitors of metalloproteinases (TIMP-1 and -2) and the activity of cathepsin C in tissue extract supernatants and their corresponding gingival crevicular fluid samples collected from periodontitis-affected and healthy patients. Our results supported evidence that tissue destruction results from an imbalance of metalloproteinases over their tissue inhibitors rather than from a sudden activation of the pro-forms of these enzymes. A significant reduction in the activity of cathepsin C also contributed to the degradative process. PMID- 11876271 TI - Influence of mastication on gastric emptying. AB - The role of mastication on digestion efficiency remains to be demonstrated. This study investigates whether masticatory function influences gastric emptying rate. Twelve normal volunteers were studied on two occasions after ingestion of the same test meal containing ham cubes, crackers, and egg (mixed with 13C-octanoic acid), chewed, in random order, either with 50 masticatory cycles or with 25 cycles, swallowing ham cubes whole. Lag phase (Tlag) and gastric half-emptying time (T1/2) were measured by means of the 13C-octanoic acid breath test. Trituration performance was assessed by the sieve test, and was expressed as the percentage of ham particles < or = 1 mm after 50 masticatory cycles. Tlag and T1/2 were significantly shorter when the meal was chewed with 50 cycles than with 25 cycles (Tlag 25.9+/-3.8 vs. 36.4+/-4.1 min, p=0.017; T1/2 49.1+/-5.7 vs. 62.5+/-6 min, p=0.009). Trituration performance was inversely related to both Tlag (r=0.621, p=0.031) and T1/2 (r=0.699, p=0.012). Comminution of food influences significantly gastric emptying rates. PMID- 11876272 TI - Beta-galactosidase activity in saliva is associated with oral malodor. AB - Deglycosylation of oral mucins may be a critical initial step leading to their subsequent proteolysis and putrefaction. The present study was undertaken to determine whether activity in saliva of a major glycosidic enzyme (beta galactosidase) is associated with oral malodor in a group of 64 subjects. Enzyme activity was detected by the use of a chromogenic substrate (X-Gal) impregnated on paper discs. Malodor-related measurements included two odor judges' assessments of whole-mouth and tongue malodor, and volatile sulfide levels measured by a portable sulfide monitor (Interscan Corp.). Beta-galactosidase assay scores were significantly associated with both odor judges' scores for whole-mouth (p < or = 0.002; Spearman) and tongue malodor (p < or = 0.001; Spearman). Beta-galactosidase activity and sulfide monitor measurements both factored significantly into multiple regression equations for odor judge scores, yielding multiple r-values ranging from 0.47 (p = 0.0007) to 0.60 (p < 0.0001). Analysis of the data presented indicates that beta-galactosidase activity in saliva is correlated with oral malodor. PMID- 11876273 TI - Pre-existing cardiovascular disease and periodontitis: a follow-up study. AB - Periodontal infections in individuals with pre-existing heart disease are believed to increase the risk for future coronary heart disease (CHD) events. The goal of this study was to search for an association between periodontitis and CHD events among individuals with pre-existing heart disease, reported in the First National Health and Nutrition Examination Survey Epidemiologic Follow-up Study. Dentate adults (n = 636) with a history of pre-existing cardiovascular disease were followed for CHD events. The presence of periodontitis and gingivitis did not increase CHD risk among these at-risk individuals (hazard ratio [HR], 0.97, and 95% confidence interval [CI], 0.72-1.31; and HR, 1.09, and 95% CI, 0.79-1.50, respectively). When limited to individuals with a self-reported prior heart attack, periodontitis was associated with a 34% decreased CHD risk (HR, 0.66; 95% CI, 0.42-1.05). It is concluded that periodontitis or gingivitis does not elevate CHD risk among individuals with a prior heart attack or self-reported pre existing cardiovascular disease. PMID- 11876275 TI - Tropoelastin expression by periodontal fibroblasts. AB - Elastic system fibers are load-bearing proteins found in periodontal tissue. There are three types--oxytalan, elaunin, and elastic fibers--which differ in their relative microfibril and elastin contents. Oxytalan fibers are known to be distributed in the periodontal ligaments and gingiva, whereas elaunin and elastic fibers are present only in the gingiva. We examined gene expression and accumulation of tropoelastin in the cell-matrix layers of human gingival fibroblasts (HGF) and periodontal ligament fibroblasts (HPLF) in vitro. HGF and HPLF were cultured in MEM containing 10% newborn calf serum for 8 wks. Northern blotting and RT-PCR analyses showed that only HGF expressed mRNA encoding tropoelastin. Western blotting analysis demonstrated 77-kDa protropoelastin and 68-kDa tropoelastin only in the cell-matrix layer of HGF cultured for 8 wks. These results suggest that the different tropoelastin expression patterns reflect the difference between HGF and HPLF phenotypes. PMID- 11876274 TI - Factors associated with delay in the diagnosis of oral cancer. AB - Early detection and treatment improve the prognosis for oral cancer. Delays from the onset of symptoms to clinical diagnosis are common. Our aim is to identify factors associated with this delay. Between 1995 and 1998, we interviewed 105 consecutive patients with histologically confirmed oral cancer in Greece. If 21 or more days elapsed from the time the patient noticed major symptoms to a definitive diagnosis, we called it a delay (52% of cases). We used logistic and linear regression to estimate odds ratios of delayed diagnosis and to identify correlates of length of delay, respectively. Former smokers had a 4.3 times greater risk of delayed diagnosis compared with current smokers (95% confidence interval: 1.1-17.1). The length of delay was greater among single patients, non smokers, or those with stage IV tumors. Clinicians should be advised that delay in the diagnosis of oral cancer occurs frequently, even in individuals who do not smoke heavily. PMID- 11876276 TI - Alpha-smooth-muscle actin in and contraction of porcine dental pulp cells. AB - The finding of expression of a muscle actin isoform, alpha-smooth-muscle actin (SMA), by the stromal cells of several tissues prompted this study of SMA expression by cells derived from the porcine dental pulp. The SMA content of the cells increased with time in culture. These SMA-containing cells were found to have the capability to contract a collagen-glycosaminoglycan analog of extracellular matrix in vitro. PMID- 11876277 TI - Experimentally induced unilateral tooth loss: histochemical studies of the temporomandibular joint. AB - Occlusal abnormality may contribute to osteoarthrosis of the temporomandibular joint (TMJ). Since mechanical force may induce changes in the extracellular matrix, we tested the hypothesis that unilateral removal of teeth and the resulting unilateral mastication change the content of glycosaminoglycans (GAGs) in the TMJ condyle and disc. Lower-right-side teeth were extracted from 12 adult male rabbits, which were killed 3 or 6 weeks later. Three normal rabbits served as controls. Sections were analyzed for morphological changes and levels of sulfated GAGs in the condyle and disc. Unilateral removal of teeth led to thickening of the condylar cartilage, alterations in the morphology of chondrocyte nuclei in the condylar cartilage and disc, and increases in levels of negatively charged ions in the hypertrophic layer of condylar cartilage. Small differences were observed, after unilateral removal of teeth, between the functional and non-functional sides of the TMJ. The results suggest that in response to mechanical stress, chondrocytes alter sulfated glycosaminoglycan (GAGs) synthesis and degradation rates, resulting in an elevated level of sulfated GAGs in the condylar cartilage. PMID- 11876278 TI - Fourier analysis reveals increased trabecular spacing in sickle cell anemia. AB - Sickle cell anemia may expand marrow spaces in the jaws. Fourier analysis is well suited to the analysis of trabecular spacing in radiographs. We hypothesize that individuals with sickle cell anemia demonstrate increased intertrabecular spacing. Periapical radiographs of 18 African Americans with sickle cell disease and 18 controls were examined by one-dimensional discrete Fourier analyses in both jaws for measurement of the spatial frequency distribution of repeating trabecular structures. A strut analysis of trabeculae was also performed and the results compared. Trabecular structures in individuals with sickle cell anemia revealed increased intertrabecular distance compared with controls. Strut analysis revealed significant reductions in trabecular complexity. Fourier analysis allows for the classification of subjects with 94% sensitivity and specificity. Fourier analysis of dental radiographs is a more effective method of identifying individuals with sickle cell anemia than strut analysis. PMID- 11876279 TI - Antiquity. PMID- 11876280 TI - Titanium elastic nails for pediatric femur fractures: lessons from the learning curve. AB - Traditionally, the treatment of choice in managing pediatric femur fractures has been traction and casting. Newer methods have focused on earlier mobility and shorter hospitalization. Use of retrograde titanium elastic nails (TENs) can quicken stabilization while allowing enough motion at the fracture site to generate excellent callus. Since TENs were first introduced in North America, our Level 1 Pediatric Trauma Center has prospectively followed all of its TEN patients. In this article, we present lessons from the learning curve of our first 50 cases--focusing on complications and their prevention. In the course of obtaining predominately excellent results, we have learned several important principles regarding TEN preoperative planning, operative technique, and aftercare. The most common problem encountered has been irritation at the nail insertion site (18% of cases). Very proximal fractures may be more challenging; unstable fractures and fractures in larger, older children are best managed with a short period of adjunctive immobilization. PMID- 11876281 TI - Percutaneous pinning of talar neck fractures. AB - In this article, we describe a technique for percutaneous pinning of talar neck fractures. This technique, which uses 2 partially threaded cannulated screws placed in opposite directions and parallel to each other, minimizes soft-tissue injury and blood-supply disruption and can shorten surgery. In addition, it allows for fracture compression and optimal positioning of screws, which increase the biomechanical stability of the fixation. PMID- 11876283 TI - Serous flexor tenosynovitis as an associated finding in meningococcal septic polyarthritis. AB - Flexor tenosynovitis complicating meningococcal septic polyarthritis may be sterile and could be misdiagnosed as disseminated gonococcal infection. Awareness of clinical findings associated with meningococcal polyarthritis is recommended to prevent onset of fulminant meningococcemia (Waterhouse-Friderichsen syndrome). PMID- 11876282 TI - Isolated posttraumatic posterior dislocation of the radial head in an adult. AB - Isolated posterior dislocation of the radial head without concomitant ulnar fracture or humeroulnar subluxation is a rare injury with few reports in the literature. We describe a case of a 27-year-old man who sustained a traction injury to the right elbow and crush injury to the forearm with subsequent posterior dislocation of the radial head. The dislocation was treated by manual reduction under anesthesia, surgical repair of the annular ligament, and subsequent rehabilitation for 6 weeks in a range-of-motion brace with the forearm in pronation. To our knowledge, this is the only case to be treated this way. Our patient reports excellent range of motion and strength 3 years after surgery. PMID- 11876284 TI - Effect of aging on male and female master athletes' performance in strength versus endurance activities. AB - Aging has traditionally been associated with several functional declines within the musculoskeletal system. In this study, we characterized and compared age related changes in performance in power and endurance activities (ie, power lifting, stationary rowing). From age 25 to age 85, men's and women's rowing performance decreases 29%; from age 25 to age 55, men's rowing performance decreases 0.12% per year, and women's performance decreases 0.23% per year. The pattern of decline in power-lifting performance is characterized by earlier onset and more rapid progression. During the fourth decade, men's and women's power lifting performance decreases 3% per year; thereafter, their performance decreases 1% per year. The difference between the age-related change rates for rowing and power-lifting is statistically significant (P = .05). The older athletes' performance in both activities suggests that the capacity for significant function in activities requiring power and endurance persists throughout life in individuals who train and who remain in good health. That strength decreases more rapidly than endurance capacity points to the importance of resistance exercises for minimizing muscle weakness in an aging population. PMID- 11876285 TI - Radio-frequency heating of the cornea: theoretical model and in vitro experiments. AB - We present a theoretical model for the study of cornea heating with radio frequency currents. This technique is used to reshape the cornea to correct refractive disorders. Our numerical model has allowed the study of the temperature distributions in the cornea and to estimate the dimensions of the lesion. The model incorporates a fragment of cornea, aqueous humor, and the active electrode placed on the cornea surface. The finite element method has been used to calculate the temperature distribution in the cornea by solving a coupled electric-thermal problem. We analyzed by means of computer simulations the effect of: a) temperature influence on the tissue electrical conductivity; b) the dispersion of the biological characteristics; c) the anisotropy of the cornea thermal conductivity; d) the presence of the tear film; and e) the insertion depth of the active electrode in the cornea, and the results suggest that these effects have a significant influence on the temperature distributions and thereby on the lesion dimensions. However, the cooling of the aqueous humor in the endothelium or the realistic value of the cornea curvature did not have a significant effect on the temperature distributions. An experimental model based on the lesions created in rabbit eyes has been used in order to compare the theoretical and experimental results. There is a tendency toward the agreement between experimental and theoretical results, although we have observed that the theoretical model overestimates the lesion dimension. PMID- 11876286 TI - On modeling the Wilson terminal in the boundary and finite element method. AB - In clinical electrocardiography, the zero-potential is commonly defined by the Wilson central terminal. In the electrocardiographic forward and inverse problem, the zero-potential is often defined in a different way, e.g., by the sum of all node potentials yielding zero. This study presents relatively simple to implement techniques, which enable the incorporation of the Wilson Terminal in the boundary element method (BEM) and finite element method (FEM). For the BEM, good results are obtained when properly adopting matrix deflation for modeling the Wilson terminal. Applying other zero-potential-definitions, the obtained solutions contained a remarkable offset with respect to the reference defined by the Wilson terminal. In the inverse problem (nonlinear dipole fit), errors introduced by an erroneous zero-potential-definition can lead to displacements of more than 5 mm in the computed dipole location. For the FEM, a method similar to matrix deflation is proposed in order to properly consider the Wilson central terminal. The matrix obtained from this manipulation is symmetric, sparse and positive definite enabling the application of standard FEM-solvers. PMID- 11876287 TI - A quality-on-demand algorithm for wavelet-based compression of electrocardiogram signals. AB - For the compression of medical signals such as electrocardiogram (ECG), excellent reconstruction quality of a highly compressed signal can be obtained by using a wavelet-based approach. The most widely used objective quality criterion for the compressed ECG is called the percent of root-mean-square difference (PRD). In this paper, given a user-specified PRD, an algorithm is proposed to meet the PRD demand by searching for an appropriate bit rate in an automatic, smooth, and fast manner for the wavelet-based compression. The bit rate searching is modeled as a root-finding problem for a one-dimensional function, where an unknown rate distortion curve represents the function and the desired rate is the root to be sought. A solution derived from root-finding methods in numerical analysis is proposed. The proposed solution is incorporated in a well-known wavelet-based coding strategy called set partitioning in hierarchical trees. ECG signals taken from the MIT/BIH database are tested, and excellent results in terms of convergence speed, quality variation, and coding performance are obtained. PMID- 11876288 TI - An averaging two-electrode probe for monitoring changes in myocardial conductivity evoked by ischemia. AB - This paper considers the applicability of effective conductivity measurements for monitoring physiological and/or pathological phenomena induced by ischemia in the myocardium. The sensitivity of a probe, calculated by means of the finite element method, to changes in the conductivity of the tissue examined is defined for this purpose. Probes developed by Schafer and collaborators (1995) and in our own departments have been examined on the basis of this sensitivity. Theoretical results were verified experimentally using a tank, enlarged models of the probes, and a specially developed electronic circuit. It follows from this study that the probe developed by Schafer et al. is characterized by positive and negative sensitivity. This can lead to misinterpretation of the measurements obtained. In contrast, the sensitivity of our probe is dominantly positive. An example of the in vivo result obtained during experimentally induced ischemia in a swine heart is included. PMID- 11876289 TI - Using electrical impedance to predict catheter-endocardial contact during RF cardiac ablation. AB - During radio-frequency (RF) cardiac catheter ablation, there is little information to estimate the contact between the catheter tip electrode and endocardium because only the metal electrode shows up under fluoroscopy. We present a method that utilizes the electrical impedance between the catheter electrode and the dispersive electrode to predict the catheter tip electrode insertion depth into the endocardium. Since the resistivity of blood differs from the resistivity of the endocardium, the impedance increases as the catheter tip lodges deeper in the endocardium. In vitro measurements yielded the impedance depth relations at 1, 10, 100, and 500 kHz. We predict the depth by spline curve interpolation using the obtained calibration curve. This impedance method gives reasonably accurate predicted depth. We also evaluated alternative methods, such as impedance difference and impedance ratio. PMID- 11876290 TI - Pacemaker interference by magnetic fields at power line frequencies. AB - Human exposure to external 50/60-Hz electric and magnetic fields induces electric fields within the body. These induced fields can cause interference with implanted pacemakers. In the case of exposure to magnetic fields, the pacemaker leads are subject to induced electromotive forces, with current return paths being provided by the conducting body tissues. Modern computing resources used in conjunction with millimeter-scale human body conductivity models make numerical modeling a viable technique for examining any such interference. In this paper, an existing well-verified scalar-potential finite-difference frequency-domain code is modified to handle thin conducting wires embedded in the body. The effects of each wire can be included numerically by a simple modification to the existing code. Results are computed for two pacemaker lead insertion paths, terminating at either atrial or ventricular electrodes in the heart. Computations are performed for three orthogonal 60-Hz magnetic field orientations. Comparison with simplified estimates from Faraday's law applied directly to extracorporeal loops representing unipolar leads underscores problems associated with this simplified approach. Numerically estimated electromagnetic interference (EMI) levels under the worst case scenarios are about 40 microT for atrial electrodes, and 140 microT for ventricular electrodes. These methods could also be applied to studying EMI with other implanted devices such as cardiac defibrillators. PMID- 11876291 TI - Compression depth estimation for CPR quality assessment using DSP on accelerometer signals. AB - Chest compression is a vital part of cardiopulmonary resuscitation (CPR). This paper demonstrates how the compression depth can be estimated using the principles of inertia navigation. The proposed method uses accelerometer sensors, one placed on the patient's chest, the other beside the patient. The acceleration to-position conversion is performed using discrete-time digital signal processing (DSP). Instability problems due to integration are combated using a set of boundary conditions. The proposed algorithm is tested on a mannequin in harsh environments, where the patient is exposed to external forces as in a boat or car, as well as improper sensor/patient alignment. The overall performance is an estimation depth error of 4.3 mm in these environments, which is reduced to 1.6 mm in a regular, flat-floor controlled environment. PMID- 11876292 TI - Directional disorder of ciliary metachronal waves using two-dimensional correlation map. AB - The interrelationship of cilia and the order of wave directions are important factors that determine the effectiveness of cilia to transport materials in mucociliary systems of the respiratory tract. The interrelationship of cilia and the directional disorder of ciliary metachronal wave were analyzed using digital microscopic images. The degree of synchronization between ciliary beats was determined by the correlation factor between two different spots. To find out the uniphase directions of beating cilia, principal axes of inertia were applied to the two-dimensional correlation map calculated from sequential ciliary images. The standard deviation of determined wave directions in a region of interest (ROI) was defined as a measure of metachronal wave disorder. The pooled mean of metachronal wave disorder was 23.4 +/- 8.79 degrees in ROIs of 8 microm x 8 microm and 25.4 +/- 6.46 degrees in 32 microm x 24 microm from the sphenoid sinus mucosa of five normal subjects. Our result shows that there is a considerable variation in metachronal wave directions of cilia beating on the epithelium. PMID- 11876293 TI - Comments on "Line patterns in the mosaic electric properties of human skin--a cross-correlation study". PMID- 11876294 TI - Methods for the analysis of protein-chromatin interactions. AB - The analysis of protein interactions with chromatin is vital for the understanding of DNA sequence recognition in vivo. Chromatin binding requires the interaction of proteins with DNA lying on the macromolecular protein surface of nucleosomes, a situation that can alter factor binding characteristics substantially when compared with naked DNA. It is therefore important to study these protein-DNA interactions in the context of a chromatin substrate, the more physiologically relevant binding situation. In this article we review techniques used in the investigation of protein interactions with defined nucleosomal templates. PMID- 11876295 TI - "De novo" sequencing of peptides recovered from in-gel digested proteins by nanoelectrospray tandem mass spectrometry. AB - Proteins separated by one-dimensional or two-dimensional gel electrophoresis can be digested in-gel with trypsin and the recovered peptides can be sequenced de novo using triple quadrupole or hybrid quadrupole time-of-flight instruments equipped with a nanoelectrospray ion source. The peptide sequences determined provide useful information for identification of proteins by homology searching for cloning of the cognate genes by PCR based approaches. PMID- 11876297 TI - Large-scale extraction of proteins. AB - The production of foreign proteins using selected host with the necessary posttranslational modifications is one of the key successes in modern biotechnology. This methodology allows the industrial production of proteins that otherwise are produced in small quantities. However, the separation and purification of these proteins from the fermentation media constitutes a major bottleneck for the widespread commercialization of recombinant proteins. The major production costs (50-90%) for typical biological product resides in the purification strategy. There is a need for efficient, effective, and economic large-scale bioseparation techniques, to achieve high purity and high recovery, while maintaining the biological activity of the molecule. Aqueous two-phase systems (ATPS) allow process integration as simultaneously separation and concentration of the target protein is achieved, with posterior removal and recycle of the polymer. The ease of scale-up combined with the high partition coefficients obtained allow its potential application in large-scale downstream processing of proteins produced by fermentation. The equipment and the methodology for aqueous two-phase extraction of proteins on a large scale using mixer-settlerand column contractors are described. The operation of the columns, either stagewise or differential, are summarized. A brief description of the methods used to account for mass transfer coefficients, hydrodynamics parameters of hold-up, drop size, and velocity, back mixing in the phases, and flooding performance, required for column design, is also provided. PMID- 11876298 TI - Immunoaffinity chromatography. AB - Immunoaffinity chromatography is a process in which the binding affinity of an antigen to a parent antibody is utilized as a basis of separation. Owing to the customized avidity and specificity, monoclonal antibodies (Mabs) have become indispensable for both protein characterization and purification. The immunosorbent performance is dependent on the support matrix upon which the antibody is immobilized and on the activation chemistry used couple the antibody to the matrix. This report details, protocols to immobilize Mabs on commercially available supports, and a method to compute immunosorbent efficiency. PMID- 11876296 TI - Classification of protein folds. AB - Classification is central to many studies of protein structure, function, and evolution. This article presents a strategy for classifying protein three dimensional structures. Methods for and issues related to secondary structure, domain, and class assignment are discussed, in addition to methods for the comparison of protein three-dimensional structures. Strategies for assigning protein domains to particular folds and homologous superfamilies are then described in the context of the currently available classification schemes. Two examples (adenylate cyclase/DNA polymerase and glycogen phosphorylase/beta glucosyltransferase) are presented to illustrate problems associated with protein classification. PMID- 11876299 TI - MALDI/MS-based epitope mapping of antigens bound to immobilized antibodies. AB - Proteolytic digestion of proteins bound to immobilized antibodies, combined with matrix assisted laser desorption (MALDI) mass spectrometric identification of the affinity-bound peptides, can be a powerful technique for epitope determination. Binding of the protein to the antibody is done while the protein is in its native, folded state. A purified protein is not required for this procedure, because only proteins containing the antigenic determinant will bind to the antibody in the initial step. The method makes use of the resistance of the antibody to enzymatic digestion. Enzymatic cleavage products of the antigenic protein not containing the epitope are washed off the beads, leaving the epitope containing fragments affinity bound to the immobilized antibody. Dissociation of the antigen-antibody complex prior to mass spectrometric analysis is unnecessary because the affinity-bound peptides are released by the MALDI matrix crystallization process, although the antibody remains covalently attached to the sepharose beads. This epitope-mapping protocol has been used in the determination of both continuous and discontinuous epitopes on both glycosylated and unglycosylated proteins. PMID- 11876300 TI - Preparation of recombinant RNase single-chain antibody fusion proteins. AB - This article describes the construction, expression, and purification of RNase single-chain antibody fusion proteins. To construct a fusion protein, the gene for each moiety, the RNase and the binding ligand, is modified separately to contain complementary DNA encoding a 13 amino acid spacer that separates the RNase from the binding moiety. Appropriate restriction enzyme sites for cloning into the vector are also added. The modified DNA is combined and fused using the PCR technique of splicing by overlap extension (1). The resulting DNA construct is expressed in inclusion bodies in BL21(DE3) bacteria that are specifically engineered for the expression of toxic proteins (2). After isolation and purification of the inclusion bodies, the fusion protein is solubilized, denatured, and renatured. The renatured RNase fusion protein mixture is purified to homogeneity by two chromatography steps. The first column, a CM-Sephadex C-50 or a heparin Sepharose column, eliminates the majority of contaminating proteins while the second column, an affinity column (Ni2+-NTA agarose), results in the final purification of the RNase fusion protein. PMID- 11876301 TI - Dye-ligand affinity adsorbents for enzyme purification. AB - Affinity chromatography is widely employed in laboratory and large-scale for the purification of biotherapeutics and diagnostics. Some of the most widely used ligands in affinity chromatography have been several reactive chlorotriazine dyes. In particular, immobilized anthraquinone dyes have found a plethora of applications in affinity chromatography because they are inexpensive, are resistant to chemical and biological degradation, are sterilizable and cleanable in situ, and are readily immobilized to generate affinity adsorbents which display high binding capacity for a broad spectrum of proteins. This article provides detailed protocols on the preparation of a dye-ligand affinity adsorbent. Also, detailed protocols for effective application of these media, emphasizing binding and elution conditions are presented. PMID- 11876302 TI - Aqueous two-phase extraction and purification of animal proteins. AB - Aqueous two-phase systems provide a rapid, easily scalable method for separation of soluble proteins from insoluble materials and other undesired proteins. The method can be operated in continuous mode. It is particularly useful for animal proteins, as it overcomes difficulties of other methods in removing bulk insoluble material, while at the same time providing purification with respect to total soluble protein. This article describes the development of methods for aqueous two-phase extraction and purification of animal proteins, at both laboratory and pilot scale. The strengths, weaknesses, and possible future prospects for the method are discussed. PMID- 11876303 TI - Protein identification by in-gel digestion and mass spectrometric analysis. AB - This protocol details a method for the identification of proteins that have been separated by gel electrophoresis. In-gel digestion of the protein bands with trypsin followed by quadrupole ion-trap or other triple quadrupole mass spectrometry techniques is described. The proteins can be identified by database searching of the mass fingerprint of the intact peptides and of the characteristic fragment masses produced by tandem mass spectrometry. PMID- 11876304 TI - Histone acetylation and histone deacetylation. AB - Regulation of inflammatory gene transcription is controlled, at least in part, by the degree of local unwinding of nucleosomal DNA. This unwinding is regulated by histone acetylation--increased acetylation results in a more loosely wound structure allowing access of basal transcription factors and RNA polymerase II. In contrast hypoacetylation of histones leads to tighter winding of DNA and reduced gene transcription. In this article we describe methods for measuring the histone acetyltransferase (HAT) and deacetylase (HDAC) activity of A549 cells. We initially describe methods examine whole cell HAT and HDAC activities and subsequently describe a technique for examining HAT activity associated with a specific co-activator CBP isolated by immunoprecipitation. These methods can also be applied to protein extracts from primary cells and from biopsy samples. PMID- 11876305 TI - Ratio model serves suprathreshold color--luminance discrimination. AB - We extended earlier results [J. Opt. Soc. Am. A 16, 2625 (1999)] to examine how the responses of the three postreceptoral mechanisms are combined to subserve discrimination of suprathreshold stimuli. Test thresholds were obtained in the presence of suprathreshold pedestals selected in different quadrants of the red green/luminance and blue-yellow/luminance planes of cardinal color space. We showed that (1) test threshold was directly proportional to pedestal contrast for pedestal contrasts exceeding five times pedestal contrast threshold, and (2) there were exceptions to this proportionality, notably when the test and pedestal directions were fixed in the cardinal directions. Results support a ratio model of suprathreshold color-luminance discrimination, in which discrimination depends on a ratio of outputs of the postreceptoral mechanisms. We also observed that when test threshold was measured as a function of test color-space direction, masking by the achromatic component of the pedestal was less than that by the chromatic component. In addition, masking by a dark (negative luminance component) pedestal was lower than masking by a light (positive luminance) pedestal of a similar contrast. Our results demonstrated that (1) there is no fundamental difference between discrimination in the isoluminant and in the two chromoluminant cardinal planes, (2) there exists the possibility that discrimination in cardinal directions differs from that in noncardinal (intermediate) directions, and (3) suprathreshold discrimination of luminance differences may be more sensitive than that of chromatic differences for a given suprathreshold pedestal. PMID- 11876306 TI - Superresolution in far-field imaging. AB - We demonstrate the feasibility of achieving superresolved images by using Fourier plane phase masks and image multiplication, which together create effective point spread functions that are not positive definite and therefore cannot be created by any single Fourier plane mask in a linear system. Three different configurations were investigated, all of which gave a spatial resolution exceeding that corresponding to the full open aperture of the optical system. One price that must be paid for the superresolution is inefficient use of the light source. PMID- 11876307 TI - Description and simulation of an active imaging technique utilizing two speckle fields: root reconstructors. AB - Quasi-monochromatic light will form laser speckle upon reflection from a rough object. This laser speckle provides information about the shape of the illuminated object. Further information can be obtained if two colors of coherent light are used, provided that the colors are sufficiently close in wavelength that the interference is also measurable. It is shown that no more than two intensities of two speckle patterns and their interference are required to produce an unambiguous band-limited image of an object, to within an overall spatial translation of the image, in the absence of measurement errors and in the case where all roots of both fields and their complex conjugates are distinct. This result is proven with a root-matching technique, which treats the electric fields as polynomials in the pupil plane, the coefficients of which form the desired complex object. Several root-matching algorithms are developed and tested. These algorithms are generally slow and sensitive to noise. So motivated, several other techniques are applied to the problem, including phase retrieval, expectation maximization, and probability maximization in a sequel paper [J. Opt. Soc. Am. A 19, 458 (2002)]. The phase-retrieval and expectation-maximization techniques proved to be most effective for reconstructions of complex objects larger than 10 pixels across. PMID- 11876308 TI - Description and simulation of an active imaging technique utilizing two speckle fields: iterative reconstructors. AB - Quasi-monochromatic light will form laser speckle upon reflection from a rough object. This laser speckle provides information about the shape of the illuminated object. In a prior paper [J. Opt. Soc. Am. A 19, 444 (2002)], it was shown that two intensities of two speckle patterns and their interference are sufficient to produce an unambiguous (except for object translation) band-limited image of the object, based on a root-matching technique described therein, in the absence of measurement error and in the case of distinct roots of the field polynomials and their complex conjugates. On the other hand, algorithms based on the root-matching technique are found to be slow and sensitive to noise. So motivated, several other techniques are applied to the problem, including phase retrieval, expectation maximization, and statistical maximization. The phase retrieval and expectation-maximization techniques proved to be most effective for reconstructions of complex objects larger than 10 pixels across, and high-quality images were formed by using three independent sets of two-field data (three frames of two-wavelength data), each comprising two speckle intensity patterns and their interference. Two additional results of note are reported. First, the expectation-maximization algorithm produced relatively good images when three or more frames each of only one speckle intensity pattern (data at just one wavelength) were used and second, the phase-retrieval algorithm when only the object autocorrelation was used also produced relatively good images for the chosen test object. PMID- 11876309 TI - Theory of quantitative phase-contrast computed tomography. AB - Phase-contrast x-ray computed tomography (CT) is an emerging imaging technique that can be implemented at third-generation synchrotron radiation sources or by using a microfocus x-ray source. Promising results have recently been obtained in materials science and medicine. At the same time, the lack of a mathematical theory comparable with that of conventional CT limits the progress in this field. Such a theory is now suggested, establishing a fundamental relation between the three-dimensional Radon transform of the object function and the two-dimensional Radon transform of the phase-contrast projection. A reconstruction algorithm is derived in the form of a filtered backprojection. The filter function is given in the space and spatial-frequency domains. The theory suggested enables one to quantitatively determine the refractive index of a weakly absorbing medium from x ray intensity data measured in the near-field region. The results of computer simulations are discussed. PMID- 11876310 TI - Finite-mode analysis by means of intensity information in fractional optical systems. AB - It is shown how a coherent optical signal that contains only a finite number of Hermite-Gauss modes can be reconstructed from the knowledge of its Radon-Wigner transform-associated with the intensity distribution in a fractional-Fourier transform optical system-at only two transversal points. The proposed method can be generalized to any fractional system whose generator transform has a complete orthogonal set of eigenfunctions. PMID- 11876311 TI - Generalized huygens-fresnel diffraction integral for misaligned asymmetric first order optical systems and decentered anisotropic Gaussian Schell-model beams. AB - The generalized Huygens-Fresnel diffraction integral for misaligned asymmetric first-order optical systems is derived by using the canonical operator method, which enables us to study propagation properties of anisotropic Gaussian Schell model (AGSM) beams through misaligned asymmetric first-order optical systems. It is shown that under the action of misaligned asymmetric first-order optical systems AGSM beams do not preserve the closed property. Therefore generalized partially coherent anisotropic Gaussian Schell-model beams called decentered anisotropic Gaussian Schell-model (DAGSM) beams are introduced, and AGSM beams can be regarded as a special case of DAGSM beams. PMID- 11876312 TI - Design of diffractive axicons producing uniform line images in Gaussian Schell model illumination. AB - We present a design method for diffractive axicons in spatially partially coherent Gaussian Schell-model illumination. The method of stationary phase applied to the Fresnel diffraction integral for on-axis intensity leads, on requiring a uniform axial image profile, to a second-order differential equation for the optimal axicon phase function. The first integral can be formally performed, and the phase function is subsequently obtained numerically. The correctness of the synthesized phase profiles is confirmed by numerical simulations using partially coherent Fresnel diffraction theory. The effects of input-beam spot size and coherence width are assessed, and influences of different forms of apodization, including asymmetric functions for narrow incident beams, in annular-aperture diffractive axicons are examined. PMID- 11876313 TI - Modal decomposition of partially coherent beams using the ambiguity function. AB - Phase-space representations of optical beams such as the ambiguity function or the Wigner distribution function have recently gained considerable importance for the characterization of coherent and partially coherent beams. There is growing interest in beam properties such as the beam propagation factor and the coherence and phase information that can be extracted from these phase-space representations. A method is proposed to decompose a partially coherent beam into Hermite-Gaussian modes by using the ambiguity function. The modal weights and the possible phase relations of the Hermite-Gaussian modes are retrieved. The method can also be applied for the decomposition of the Wigner distribution function. Some examples are discussed in the scope of beam characterization. PMID- 11876314 TI - Electromagnetic-wave scattering by a sphere with multiple spherical inclusions. AB - An exact solution to the problem of electromagnetic-wave scattering from a sphere with an arbitrary number of nonoverlapping spherical inclusions is obtained by use of the indirect mode-matching technique. A set of linear equations for the wave amplitudes of the electric field intensity throughout the inhomogeneous sphere and in the surrounding empty space is determined. Numerical results are calculated by truncation and matrix inversion of that set of equations. Specific information about the truncation number pertaining to the multipole expansions of the electric field intensity is given. The theory and the accompanying computer code successfully reproduce the results of other pertinent papers. Some numerical results [Borghese et al., Appl. Opt. 33, 484 (1994)] were not reproduced well, and that discrepancy is discussed. Our numerical investigation is focused on an acrylic sphere with up to four spherical inclusions. This is the first time that numerical results are presented for a sphere with more than two spherical inclusions. Interesting remarks are made about the effect that the look direction and the structure of the inhomogeneity have on backscattering by the acrylic host sphere. PMID- 11876315 TI - Coupled multipolar interactions in small-particle metallic clusters. AB - We propose a new formalism for computing the optical properties of small clusters of particles. It is a generalization of the coupled dipole-dipole particle interaction model and allows one in principle to take into account all multipolar interactions in the long-wavelength limit. The method is illustrated by computations of the optical properties of N = 6 particle clusters for different multipolar approximations. We examine the effect of separation between particles and compare the optical spectra with the discrete-dipole approximation and the generalized Mie theory. PMID- 11876316 TI - Diffusion equation boundary conditions for the interface between turbid media: a comment. AB - This discussion reconciles differences in literature expressions for the diffusion approximation boundary conditions for the interface between two turbid media with different refractive indices. PMID- 11876317 TI - Can simple particle shapes be used to model scalar optical properties of an ensemble of wavelength-sized particles with complex shapes? AB - We compute the scalar optical properties of size-shape distributions of wavelength-sized randomly oriented homogeneous particles with different nonaxially symmetric geometries and investigate how well they can be modeled with a simple spherical, spheroidal, or cylindrical particle model. We find that a spherical particle model can be used to determine the extinction and scattering cross sections, the single-scattering albedo, and the asymmetry parameter with an error of less than 2%, whereas the extinction-to-backscatter ratio Reb is reproduced only with an error of 9%. The cylindrical and spheroidal particle models yield slightly improved results for Reb that deviate from those obtained for the complex particle ensemble by 7% and 5%, respectively. Large discrepancies between results of the different models are observed for the linear depolarization ratio, thus indicating limitations of models based on simple particle shapes. PMID- 11876318 TI - Analysis of time-gated imaging through scattering media by a Fourier optics approach. AB - The method of Fourier optics is applied to the problem of time-gated imaging through scattering media. Tb adapt the problem to this treatment, appropriate alterations are made: The continuous medium is replaced by a cascade of thin scatterers, and a spatial filtering process is substituted for the conventional gating processes. Closed-form solutions are derived. PMID- 11876319 TI - Multiconjugate adaptive optics for large telescopes: analytical control of the mirror shapes. AB - We present an analytical algorithm for deriving the shapes of the deformable mirrors to be used for multiconjugate adaptive correction on a large telescope. The algorithm is optimal in the limit where the overlap of the wave-front contributions from relevant atmospheric layers probed by the guide stars is close to the size of the pupil. The fundamental principle for correction is based on a minimization of the sum of the residual power spectra of the phase fluctuations seen by the guide stars after correction. On the basis of the expressions for the mirror shapes, so-called layer transfer functions describing the distribution of the correction of a single atmospheric layer among the deformable mirrors and the resulting correction of that layer have been derived. It is shown that for five guide stars distributed in a regular cross, two- and three-mirror correction will be possible only up to a maximum frequency defined by the largest separation of the conjugate altitudes of the mirrors and by the angular separation of the guide stars. The performance of the algorithm is investigated in the K band by using a standard seven-layer atmosphere. We present results obtained for two guide-star configurations: a continuous distribution within a given angular radius and a five-star cross pattern with a given angular arm length. The wave-front fluctuations are subjected to correction using one, two, and three deformable mirrors. The needed mirror dynamic range is derived as required root-mean-square stroke and actuator pitch. Finally the performance is estimated in terms of the Strehl ratio obtained by the correction as a function of field angle. No noise has been included in the present analysis, and the guide stars are assumed to be at infinity. PMID- 11876320 TI - Invariant identification of material mixtures in airborne spectrometer data. AB - We present an algorithm for identifying linear mixtures of a specified set of materials in 0.4-2.5 microm airborne imaging spectrometer data. The algorithm is invariant to the illumination and atmospheric conditions and the relative amounts of the specified materials within a pixel. Only the spectral reflectance functions for the specified materials are required by the algorithm. Invariance over illumination and atmospheric conditions is achieved by incorporating a physical model for scene variability in the constrained optimization formulation. The algorithm also computes estimates of the amounts of the specified materials in identified mixtures. We demonstrate the effectiveness of the algorithm by using real and synthetic Hyperspectral Digital Imaging Collection Experiment imagery acquired over a range of conditions and altitudes. PMID- 11876321 TI - Inverse problem in optical diffusion tomography. II. Role of boundary conditions. AB - We consider the inverse problem of reconstructing the absorption and diffusion coefficients of an inhomogeneous highly scattering medium probed by diffuse light. The role of boundary conditions in the derivation of Fourier-Laplace inversion formulas is considered. Boundary conditions of a general mixed type are discussed, with purely absorbing and purely reflecting boundaries obtained as limiting cases. Four different geometries are considered with boundary conditions imposed on a single plane and on two parallel planes and on a cylindrical and on a spherical surface. PMID- 11876322 TI - Optimum divergence angle of a Gaussian beam wave in the presence of random jitter in free-space laser communication systems. AB - The average bit error rate (BER) of optical communication systems is considered in the presence of random angular jitter. First, the received power and the BER in the absence of jitter are reviewed. Then the average BER is obtained in the presence of circularly symmetric, normally distributed jitter by using the probability density function of the optical signal. By minimizing the power penalty for average BER, the optimum ratio of the divergence angle of the laser beam to the random angular jitter at the desired BER is obtained. An analytic approximation of the optimum ratio is derived as a function of the desired average BER. The results can be used for designing the link budget of optical communication and tracking channels in the presence of jitter. PMID- 11876323 TI - Theory of near-field magneto-optical imaging. AB - Scanning near-field optical microscopy has been recently applied to the imaging of magnetic samples. It was shown experimentally that an apertureless microscope suffers a substantial loss of resolution when used for magneto-optical imaging compared with that for conventional imaging. No such change is observed for aperture microscopes. We explain this observation by developing a model for the imaging process that incorporates the response of the probe. We calculate real observable properties such as the rotation of polarization at the detector or the circular dichroism signal and thus simulate magneto-optical images of a domain structure in cobalt for both aperture and apertureless microscopes. PMID- 11876324 TI - New class of axially apodizing filters for confocal scanning microscopy. AB - A new class of axially apodizing continuously transmitting filters is analytically determined and numerically implemented. The longitudinal and lateral properties of the associated point-spread function are displayed both for a single filtered lens and for a combination with a laterally superresolving mask in a confocal configuration. Interesting imaging properties relative to the increase in axial contrast and transverse resolving power for dephasing ring-free pupil filters of moderate losses in luminosity are predicted. PMID- 11876325 TI - Tailored freeform optical surfaces. AB - Freeform optical surfaces embedded in three-dimensional space, without any symmetry, are tailored so as to solve the archetypal problem of illumination design: redistribute the radiation of a given small light source onto a given reference surface, thus achieving a desired irradiance distribution on that surface. The shape of the optical surface is found by solving a set of partial nonlinear differential equations. For most cases, a few topologically distinct solutions exist, given suitable boundary conditions. PMID- 11876326 TI - Use of single-mode waveguides to correct the optical defects of a nulling interferometer. AB - In the area of long-baseline nulling interferometry, high rejection ratios are needed to cancel out the light of a bright central source and look for nearby faint structures or companions. These rejection requirements directly translate into drastic optical constraints. We discuss the possibility of using single-mode waveguides for such applications. Conversely to simple pinholes, single-mode waveguides efficiently correct wave-front defects of both high- and low-order spatial frequencies, ensure a perfect matching of the amplitude profiles coming from the various beams, and can be used with almost optimum coupling efficiency over a broad optical bandpass (typically an octave). They then appear to greatly enhance the feasibility of high-dynamic interferometric coronagraphs. PMID- 11876327 TI - Understanding multilayers from a geometrical viewpoint. AB - We reelaborate on the basic properties of lossless multilayers. We show that the transfer matrices for these multilayers have essentially the same algebraic properties as the Lorentz group SO(2, 1) in a (2 + 1)-dimensional space-time as well as the group SL(2, R) underlying the structure of the ABCD law in geometrical optics. By resorting to the Iwasawa decomposition, we represent the action of any multilayer as the product of three matrices of simple interpretation. This group-theoretical structure allows us to introduce bilinear transformations in the complex plane. The concept of multilayer transfer function naturally emerges, and its corresponding properties in the unit disk are studied. We show that the Iwasawa decomposition is reflected at this geometrical level in three simple actions that can be considered the basic pieces for a deeper understanding of the multilayer behavior. We use the method to analyze in detail a simple practical example. PMID- 11876328 TI - Analysis of optical waveguide structures by use of a combined finite difference/finite-difference time-domain method. AB - We present a method for full-wave characterization of optical waveguide structures. The method computes mode-propagation constants and cross-sectional field profiles from a straight forward discretization of Maxwell's equations. These modes are directly excited in a three-dimensional finite-difference time domain simulation to obtain optical field transmission and reflection coefficients for arbitrary waveguide discontinuities. The implementation uses the perfectly-matched-layer technique to absorb both guided modes and radiated fields. A scattered-field formulation is also utilized to allow accurate determination of weak scattered-field strengths. Individual three-dimensional waveguide sections are cascaded by S-parameter analysis. A complete 10(4)-section Bragg resonator is successfully simulated with the method. PMID- 11876330 TI - Standards of practice for assessment of aspiration of enteral tube feedings in patients with artificial airways. PMID- 11876329 TI - Calculated impact of higher-order monochromatic aberrations on retinal image quality in a population of human eyes. AB - We calculated the impact of higher-order aberrations on retinal image quality and the magnitude of the visual benefit expected from their correction in a large population of human eyes. Wave aberrations for both eyes of 109 normal subjects and 4 keratoconic patients were measured for 3-, 4-, and 5.7-mm pupils with a Shack-Hartmann sensor. Retinal image quality was estimated by means of the modulation transfer function (MTF) in white light. The visual benefit was calculated as the ratio of the MTF when the monochromatic higher-order aberrations are corrected to the MTF corresponding to the best correction of defocus and astigmatism. On average, the impact of the higher-order aberrations for a 5.7-mm pupil in normal eyes is similar to an equivalent defocus of approximately 0.3 D. The average visual benefit for normal eyes at 16 c/deg is approximately 2.5 for a 5.7-mm pupil and is negligible for small pupils (1.25 for a 3-mm pupil). The benefit varies greatly among eyes, with some normal eyes showing almost no benefit and others a benefit higher than 4 at 16 c/deg across a 5.7-mm pupil. The benefit for keratoconic eyes is much larger. The benefit at 16 c/deg is 12 and 3 for 5.7- and 3-mm pupils, respectively, averaged across four keratoconics. These theoretical benefits could be realized in normal viewing conditions but only under specific conditions. PMID- 11876331 TI - Nurturing unborn baby strains nurses' emotions. PMID- 11876334 TI - Primary nursing for neonates. PMID- 11876335 TI - Carotid endarterectomy: creating the pathway to 1-day stay. AB - Use of a collaborative team approach to design a care pathway and standing orders for carotid endarterectomy patients achieved the project's goals. Variation, LOS, and resource consumption were decreased while quality of care and patients' satisfaction levels were maintained. Education of patients, patients' families, and staff members increased. Coordination between caregivers increased. The consistent concurrent database provided a feedback loop for continued change and for setting the target. Essential to the success was sponsorship from key leadership via the hospital's steering committee. The diverse membership of key associates on the CQI team helped to create an excellent revised carotid endarterectomy process and ensured full implementation. This membership of the CQI team was essential to comprehensive education and implementation. The step-by step implementation kept the project moving forward. Creating a care pathway and changing practice require collaboration between nurses, doctors, and administrators. Creativity and systematic, thorough steps are what move a practice change from idea to inception. PMID- 11876336 TI - Endovascular grafting for repair of abdominal aortic aneurysm. PMID- 11876338 TI - Internet relay and Web-based chat: powerful tools for teaching and collegial networking. PMID- 11876337 TI - Providing spiritual care to cardiac patients: assessment and implications for practice. PMID- 11876339 TI - Stabilizing unstable plaque in acute coronary syndromes: implications for cardiac surgical patients. PMID- 11876340 TI - Managing children with splenic laceration. PMID- 11876341 TI - Family caregiving in hospice: effects on psychological and health functioning among spousal caregivers of hospice patients with lung cancer or dementia. AB - INTRODUCTION: Caregiving stress has been found to lead to depression and poor health among caregivers compared with age-matched non-caregiving controls. However, most of these studies have focused on dementia caregivers, and have not included hospice caregivers. The aim of this project was to assess the impact of caregiving stress on psychological and health functioning among spousal caregivers of hospice patients, in contrast to demographically matched non caregiving controls. A secondary aim was to compare the caregiving stressors and psychological and health functioning between spousal caregivers of hospice patients with dementia versus lung cancer. METHODS: Forty spousal caregivers of hospice patients with dementia, and 40 spousal caregivers of hospice patients with lung cancer, were compared on admission to hospice, using measures of caregiving stressors, depression, life satisfaction, and physical health, with a sample of 40 demographically equated control subjects. RESULTS: Both groups of caregivers showed higher depression, lower life satisfaction and poorer physical health (p < .05) compared with non-caregivers. Over half of all caregivers evidenced clinically significant levels of depression, with rates of depression about three times the prevalence found in community samples of older adults. CONCLUSIONS: While family caregivers of hospice patients with dementia and lung cancer face very different objective stressors, the negative psychological and health impacts on the caregiver are marked and comparable across diagnosis. Hospice family caregivers are at high risk for both psychological and physical health disorders, and caregiver depression and health problems should be systematically assessed and treated by the hospice team. PMID- 11876342 TI - Development of a home-based family caregiver cancer education program. AB - INTRODUCTION: This article describes a home-based educational program developed specifically for family caregivers of cancer patients who receive hospice and home care. The overall aim of this educational program is to specifically address family caregivers' needs for acquisition of necessary knowledge and skills to meet the physical and psychosocial demands associated with caring for a patient with advanced cancer. PROGRAM DEVELOPMENT: Originally, components of this program were offered in a small group discussion format within hospital and community settings. The educational program was transformed to accommodate the unique needs and constraints of homebound family caregivers who have very limited time and/or opportunities for support and education outside of the home. The program is comprised of educational modules that provide hospice and home care professionals with written and audiovisual materials designed to facilitate brief, structured, educational encounters with family caregivers in the home setting. DISCUSSION: Two hundred thirty-seven educational module kits were distributed to professionals affiliated with twenty-four home care and hospice agencies in the Greater Philadelphia area. Results of a telephone survey designed to elicit evaluation data from professional staff members who had used the educational modules are presented. Limitations, plans for future program evaluation, cost implications, and implementation recommendations related to this educational program are described. PMID- 11876343 TI - Junior medical students' perceptions of an introductory hospice experience. AB - INTRODUCTION: The importance of palliative care education in the medical school curriculum is becoming more recognized. The purpose of this study was to assess medical students' perceptions of an introductory hospice experience. METHODS: Forty-one second-year medical students took part in an introductory hospice experience in which they were acquainted with a wide range of hospice services provided to patients and families by an interdisciplinary team involved in hospice care. In addition, the students visited patients' homes individually with an experienced hospice nurse or social worker. At the end of their experience, the students were asked to complete a multi-item evaluation questionnaire in order to share their perceptions of the hospice experience and their suggestions for improvement of the course. RESULTS: The students spent an average of four hours on their introductory hospice experience, and they indicated that all of their personal goals for their experience had been met. Suggestions for improvement of the course were to increase the amount of course time allotted and to provide further opportunity to see more patients. Overall, the students rated their experience as "above average" to "excellent." CONCLUSIONS: According to the medical students who participated, the introductory hospice experience was a worthwhile and valuable educational experience. An equal or increased amount of hospice time should be allotted in the education of future junior medical students. PMID- 11876344 TI - Parents' perceived benefits and barriers of adolescents' diabetes self management: part 2. AB - Knowledge is limited about parents' decision making regarding adolescents' assumption of diabetes management. This study's purpose was to identify parents' perceived benefits and barriers about this process. We interviewed 17 parents of adolescents with type I diabetes. Questions were adapted from Janis and Mann's (1977) decisional balance sheet. Interviews were audiotaped and transcribed verbatim. Descriptive expressions were identified and categories inductively generated. Inter-rater agreement was initially 85% and was 100% after discussion of disagreements. Parents' perceived benefits for themselves were relief from burden and knowledge and confidence and/or pride in adolescents' abilities. Their perceived benefits for their sons and daughters were also twofold: knowledge and/or confidence in abilities and freedom, independence, and/or control. Parents perceived barriers for themselves were loss of control, authority, and supervision, dealing with consequences. and expression of lack of barriers. Their perceived barriers for their sons and daughters were burden of responsibility and also lack of barriers. Understanding parents' perceived benefits and barriers would be beneficial to health care professionals when working with parents, because decision making related to the transfer of diabetes management to their adolescent sons and daughters is crucial to the success of its management. PMID- 11876345 TI - California healthy and ready to work transition health care guide: developmental guidelines for teaching health care self-care skills to children. AB - This article presents developmental guidelines that can be used by parents and nurses for teaching children health care self-care skills. These guidelines are intended to provide a framework for instructing parents and nurses about the developmentally appropriate skills children can learn to become self sufficient in managing their own health care needs. The guidelines are based on the developmental frameworks of Piaget and Erikson and self-care models of Orem. Although general in scope, these guidelines can be individualized to the needs of each child. PMID- 11876346 TI - Fresh beginnings. PMID- 11876347 TI - Moving from technical to critical reflection in journalling: an investigation of students' ability to incorporate three levels of reflective writing. AB - This paper outlines a research project aimed at changing the levels of reflection of preregistration nursing students in a tertiary institution. Whilst reflection is widely espoused now in nursing, few studies have been found that identify whether the level of reflective writing can be identified or developed by students. Anecdotal and research evidence (Powell 1989; van Manen 1977) however indicates that most student reflective writing occurs at the technical level. A descriptive exploratory study using both qualitative and quantitative techniques was undertaken to apply van Manen's (1977) levels in a structured way in an attempt to facilitate the student's understanding and use of the levels in their reflective writing. The findings of the study indicate that student self evaluation and identification of the levels in their own writing can lead to change in the levels of critical reflective writing achieved by undergraduate students. PMID- 11876348 TI - A survey of nurses' views on indicators for continuing competence in nursing. AB - The study, commissioned by the Australian Nursing Council Inc. (ANCI), sought to develop an approach to the maintenance of continuing competence in nursing broadly acceptable to nurses in all States and Territories and included the conduct of a postal survey of registered nurses (RNs) throughout Australia. An options booklet and accompanying questionnaire were posted to a random sample of 2 % of RNs from all Sates and the Northern Territory in Australia. The sample consisted of 4,133 RNs and 1,005 completed questionnaires were returned representing a 24.3% response rate. Data were analysed using SPSSX. The results suggest that respondents most favoured the introduction of a signed declaration of competence for all nurses seeking annual relicensing and the conduct of random competency audits by nursing regulatory authorities. PMID- 11876349 TI - Factors influencing nurses' decisions to use non-pharmacological therapies to manage patients' pain. AB - This study investigated nurses' beliefs and attitudes toward the use of non pharmacological therapies as adjunct pain management strategies. Registered nurses (RNs) (n=37) from the medical, surgical, oncology/palliative care and critical care areas of two Australian hospitals participated in a series of focus group discussions that explored the use of non-pharmacological therapies to help manage patients' pain in a hospital setting. Results from the discussions identified that nurses believe non-pharmacological therapies offer several advantages to the management of patients' pain and general well being. For example non-pharmacological therapies were recognised to be useful as adjuncts while waiting for medications to take effect. However significant barriers such as lack of organisational and professional support were also identified as hindering nurses' current usage of non-pharmacological therapies. Further investigation of the key issues from this study is recommended to improve non pharmacological pain management and enhance patient outcomes. PMID- 11876350 TI - Continued neglect of rural and remote nursing in Australia: the link with poor health outcomes. AB - Difficulties facing Australian rural and remote nurses first emerged decades ago and recent Australian Federal Government rural health strategies have promised improvements in health outcomes. However closer scrutiny of the funding allocation supporting these claims reveals that although nurses provide the majority of health care, they receive only a small fraction of funding support. Successive rural health strategies have continued to overlook nurses' concerns regarding the nurse-practice environment interface. This persistent lack of political support stifles organisational support for the nursing role, resulting in frustration, resignation and diminished health care delivery. Meanwhile, rural and remote population health outcomes fail to show signs of improvement. PMID- 11876351 TI - Reflections on nursing: a time of challenge and choice. PMID- 11876352 TI - Women's experience of early miscarriage: a phenomenological study. AB - This paper describes the experience of three women who had experienced early miscarriage within the previous 12 months. Three major themes of loss emerged: the loss of a baby, the loss of the role of motherhood and the loss of the hopes and dreams the women possessed for their baby. These losses were complicated by the women's negative thoughts about the care they received while in hospital, their perceptions of health professional's negative attitudes, the lack of information given to them, and the lack of understanding shown by family and friends about their situation. The authors recommend a number of changes to practice including the need for an increased sensitivity towards women following early miscarriage. PMID- 11876353 TI - On-line fluorescence determination of pressure mediated outer membrane damage in Escherichia coli. AB - The outer membrane (OM) of Gram-negative bacteria provides a protective barrier for natural occurring inhibitors. Pressure mediated OM permeabilisation therefore contributes to the elimination of Escherichia coli and Salmonella by pressure preservation processes. Pressure mediated inactivation, sublethal injury, and membrane permeabilisation of E. coli were determined using two strains differing in their barotolerance. Pressure treatment of E. coli TMW 2.427 at 300, 500 and 600 MPa for 40 min resulted in a 0, 1, and greater 6 log decrease of viable cell counts, respectively. The kinetics of OM and cytoplasmic membrane permeabilisation after pressure treatment were determined by staining of pressure treated cells with the fluorescent dyes propidium iodide (PI) and 1-N phenylnaphtylamine (NPN), respectively. A slight increase of PI fluorescence was observed at conditions resulting in a greater 6 log decrease of viable cell counts only. In contrast, increased NPN fluorescence indicating OM permeabilisation was observed prior to cell death and sublethal injury. An on line assay for determination of pressure mediated OM damage based on NPN fluorescence was established to distinguish between reversible and irreversible OM damage. Generally, the same degree of outer membrane damage was observed by either on line or off line determinations. However, whereas reversible membrane damage occurred fast and in thermodynamic equilibrium with pressure conditions, irreversible outer membrane damage was a time dependent process. PMID- 11876354 TI - Surface immunolocalisation of HPr in the equine pathogen Streptococcus equi. AB - We have investigated the surface localisation of the phosphotransferase system protein HPr in the equine pathogen Streptococcus equi subsp. equi using immunogold localisation and transmission electron microscopy. Like the LppC acid phosphatase lipoprotein, a reference surface antigen, the S. equi HPR could be clearly detected on the surfaces of intact cells. This study is consistent with previous reports that some streptococcal HPr is cell surface associated and suggests that the extracytoplasmic mobilisation and transfer of phosphate groups by streptococci warrant further investigation. PMID- 11876355 TI - Stable low molecular weight RNA analyzed by staircase electrophoresis, a molecular signature for both prokaryotic and eukaryotic microorganisms. AB - Low-molecular weight RNA (LMW RNA) analysis using staircase electrophoresis was performed for several species of eukaryotic and prokaryotic microorganisms. According to our results, the LMW RNA profiles of archaea and bacteria contain three zones: 5S RNA, class 1 tRNA and class 2 tRNA. In fungi an additional band is included in the LMW RNA profiles, which correspond to the 5.8S RNA. In archaea and bacteria we found that the 5S rRNA zone is characteristic for each genus and the tRNA profile is characteristic for each species. In eukaryotes the combined 5.8S and 5S rRNA zones are characteristic for each genus and, as in prokaryotes, tRNA profiles are characteristic for each species. Therefore, stable low molecular weight RNA, separated by staircase electrophoresis, can be considered a molecular signature for both prokaryotic and eukaryotic microorganisms. Analysis of the data obtained and construction of the corresponding dendrograms afforded relationships between genera and species; these were essentially the same as those obtained with 16S rRNA sequencing (in prokaryotes) and 18S rRNA sequencing (in eukaryotes). PMID- 11876356 TI - Fats and fatty acids as growth factors for Lactobacillus delbrueckii. AB - The effects of various fats and fatty acids on the growth of Lactobacillus delbrueckii strains have been studied using modified MRS broth without Tween 80 as a basic growth medium. Among the six L. delbrueckii strains studied all except one strain required Tween 80 or Tween 20 as a fatty acid supplement for the growth. Tween 40 and Tween 60, which contain solely medium and long chain saturated fatty acids, inhibited the growth of all L. delbrueckii strains when present as a sole fat supplement in MRS broth. Free oleic acid but not free lauric acid could substitute Tween 80 or Tween 20 supplement suggesting that unsaturated fatty acids are essential growth factors for most L. delbrueckii strains. Among the natural food oils tested, the oils containing the lowest amounts of saturated long chain fatty acids promoted the growth of L. delbrueckii most effectively. Especially cellular C18:1 and C19 cyclopropane fatty acid contents of L. delbrueckii were strongly affected by exogenous fatty acid composition and by strain suggesting genetic diversity and polymorphism among the genes encoding and/or regulating cyclopropane synthase. In addition obviously most if not all L. delbrueckii strains lack particular synthase, desaturase and/or dehydrase activities required for de novo synthesis of long chain unsaturated fatty acids. These biochemical features could be used as informative chemotaxonomic characteristics for L. delbrueckii starter strain identification and selection. PMID- 11876357 TI - Cellular fatty acid composition from Sarcobium lyticum (Legionella lytica comb. nov.)--an intracellular bacterial pathogen of amoebae. AB - Legionella lytica comb. nov. an intracellular bacterial pathogen of small free living amoebae was subjected to cellular fatty acid (FA) analysis employing base and acid catalyzed cleavage, gas-liquid chromatography and mass spectrometry. Both unbranched and branched (iso and anteiso) FA of chains ranging from 14 to 30 carbon atoms occurred. The presence of two long-chain FA: 27-oxo-octacosanoic acid and heptacosane-1,27-dioic acid, characteristic for legionellae, was found. Nine amide-linked 3-hydroxy-FA were revealed. The main 3-hydroxy-fatty acids comprise: 3-OH-14:0, 3-OH-16:0, 3-OH-18:0, 3-OH-i18:0, 3-OH-15:OH, 3-OH-i16:0 amd 3-OH-i17:0. The profile of hydroxy FAs permits allocation of L. lytica to group 3 of legionellae which comprise blue-white fluorescent species. PMID- 11876358 TI - Gordonia namibiensis sp. nov., a novel nitrile metabolising actinomycete recovered from an African sand. AB - A polyphasic approach was used to establish the taxonomic position of two actinomycetes isolated from a Namibian soil and shown to utilise nitrile compounds as growth substrates. The organisms, strains NAM-BN063AT and NAM BN063B, had chemical and morphological properties consistent with their assignment to the genus Gordonia. Direct 165 rRNA sequencing studies confirmed the taxonomic position of the strains following the generation of phylogenetic trees using four different algorithms. The strains consistently formed a distinct phylogenetic line within the evolutionary radiation occupied by gordoniae and were most closely related to Gordonia rubropertincta DSM 43197T. DNA:DNA relatedness studies indicated that the two organisms belonged to a genomic species that was readily distinguished from G. rubropertincta. The unique phenotypic profile of the strains sharply separated them from representatives of all of the validly described species of Gordonia. The combination of genotypic and phenotypic data indicates that the two strains should be classified in the genus Gordonia as a new species. The name proposed for this taxon is Gordonia namibiensis, the type strain is NAM-BN063AT (= DSM 44568T = NCIMB 13780T). PMID- 11876359 TI - The coral bleaching Vibrio shiloi Kushmaro et al. 2001 is a later synonym of Vibrio mediterranei Pujalte and Garay 1986. AB - The coral bleaching Vibrio shiloi LMG 19703T was characterized by means of Fluorescent Amplified Fragment Length Polymorphism (FAFLP), DNA-DNA hybridisation, mol% G+C content, fatty acids methyl ester (FAME) analysis and phenotypical tests. Numerical analysis of the FAFLP band patterns indicated that the type strain of V. shiloi in fact belongs to the species V. mediterranei. The type strains of both species shared 77% DNA similarity, as determined by DNA-DNA hybridisation experiments at stringent conditions. Moreover, V. shiloi and V. mediterranei showed almost identical fatty acid composition and phenotypical features. Collectively, the genotypic and phenotypic data presented in this study suggest that V. shiloi Kushmaro et al. 2001 should be considered a later synonym of V. mediterranei Pujalte and Garay 1986. The involvement of V. mediterranei in coral bleaching was unknown until now. PMID- 11876360 TI - Genomic diversity amongst Vibrio isolates from different sources determined by fluorescent amplified fragment length polymorphism. AB - The genomic diversity among 506 strains of the family Vibrionaceae was analysed using Fluorescent Amplified Fragments Length Polymorphisms (FAFLP). Isolates were from different sources (e.g. fish, mollusc, shrimp, rotifers, artemia, and their culture water) in different countries, mainly from the aquacultural environment. Clustering of the FAFLP band patterns resulted in 69 clusters. A majority of the actually known species of the family Vibrionaceae formed separate clusters. Certain species e.g. V. alginolyticus, V. cholerae, V. cincinnatiensis, V. diabolicus, V. diazotrophicus, V. harveyi, V. logei, V. natriegens, V. nereis, V. splendidus and V. tubiashii were found to be ubiquitous, whereas V. halioticoli, V. ichthyoenteri, V. pectenicida and V. wodanis appear to be exclusively associated with a particular host or geographical region. Three main categories of isolates could be distinguished: (1) isolates with genomes related (i.e. with > or =45% FAFLP pattern similarity) to one of the known type strains; (2) isolates clustering (> or =45% pattern similarity) with more than one type strain; (3) isolates with genomes unrelated (<45% pattern similarity) to any of the type strains. The latter group consisted of 236 isolates distributed in 31 clusters indicating that many culturable taxa of the Vibrionaceae remain as yet to be described. PMID- 11876361 TI - Functional grouping of thermophilic Bacillus strains using amplification profiles of the 16S-23S internal spacer region. AB - Molecular and biochemical assays were used to determine the identification of thermophilic bacilli isolated from New Zealand milk powder. One hundred and forty one isolates of thermophilic bacilli were classified into six species using biochemical profiles. Geobacillus stearothermophilus represented 56% of the isolates. All isolates were also analysed by randomly amplified polymorphic DNA (RAPD) analysis, with 45 types identified. Amplification of the 16S-23S rDNA internal spacer region produced two to eight amplification products per strain. The patterns from gel electrophoresis of the internal spacer region amplicons formed two major groupings suggesting the possibility of two distinct species. Partial sequences of 16S rDNA from representatives from each group were compared with sequences in GeneBank and were found to match the 16S rDNA sequences of B. flavothermus and G. thermoleovorans. Primers were designed for these species and used to screen an arbitrary selection of 59 of the dairy isolates. This enabled the identification of 28 isolates as B. flavothermus and 31 isolates as Geobacillus species and these appear to be the predominant isolates in the New Zealand milk powder samples examined. Comparison of the fragment pattern generated by amplification of the 16S-23S rDNA internal spacer region is a simple method to differentiate thermophilic Bacillus species associated with the dairy industry. PMID- 11876362 TI - Differences between subcultures of the Mesorhizobium loti type strain from different culture collections. AB - Because of differences in the reported 16S rRNA gene sequence of the Mesorhizobium loti type strain available from different culture collections, we collected different subcultures of this strain and compared them by 16S rDNA sequencing, SDS-PAGE of whole-cell protein extracts and RAPD-PCR. Our results indicate that the 16S rDNA sequence differences can be explained by the presence of two different organisms in one of the subcultures. In addition, even for subcultures of the type strain that had identical 16S rDNA sequences, small differences could be observed in the protein profiles and in the RAPD-PCR patterns. These latter observations indicate that maintenance procedures necessary for long-term preservation by freeze-drying can cause subcultures of the same original strain to undergo changes, effectively leading to different fingerprints even though 16S rDNA sequences remain identical. PMID- 11876363 TI - Differentiation of Lactobacillus plantarum, L. pentosus and L. paraplantarum species by RAPD-PCR and AFLP. AB - Two high-resolution genotypic techniques (RAPD-PCR and AFLP) were evaluated for their possibility to discriminate the species Lactobacillus plantarum, Lactobacillus pentosus and Lactobacillus paraplantarum and to type these taxa at the infra-species level. In total 23 strains of L. plantarum, three strains of L. pentosus, two strains of L. paraplantarum and two related strains for which the species assignment was not clear, were studied. For RAPD-PCR, suitable oligonucleotides and amplification conditions were selected and tested. For AFLP, a double digest of total genomic DNA was used and a subset of restriction fragments was selectively amplified and visualised using different primer combinations. Both methodologies generated, species-specific electrophoretic profiles. Moreover, the presence of distinct subgroups was revealed within the species L. plantarum. PMID- 11876364 TI - Polyphasic investigation of the diversity within Lactobacillus plantarum related strains revealed two L. plantarum subgroups. AB - The diversity of 140 strains related to Lactobacillus plantarum was investigated using a polyphasic approach combining two molecular techniques: randomly amplified polymorphic DNA fingerprinting (RAPD) and Southern hybridisation with a pyr probe on BglI digests of chromosomal DNA, as well as phenotypic characterization. The RAPD technique allowed us to classify a subset of 60 representative strains into four groups. One group belonged to Lactobacillus paraplantarum, the second to Lactobacillus pentosus and the two remaining groups to L. plantarum (G(L)p1 and G(L)p2). The Southern hybridisation technique (F. Bringel, M.-C. Curk and J.-C. Hubert, Int. J. Syst. Bacteriol. 46: 588-594, 1996) revealed nine groups of profiles (I to IX). Results indicated an excellent convergence between RAPD and hybridisation classifications for more than 93% (56/60) of the strains studied. When we compared the fermentation patterns of the L. plantarum strains, three differences were found. Melezitose fermentation was not fermented by the G(L)p2 RAPD group, unlike the G(L)p1 RAPD group which included L. plantarum type strain NCIMB11974T. Second, alpha-methyl-D-mannoside was fermented by a majority of the strains of the G(L)p1 RAPD group but by none of the strains in the G(L)p2 RAPD group. Third, dulcitol was catabolized by nearly half of the strains of the G(L)p2 RAPD group but by none of the strains in the G(L)p1 RAPD group. Molecular diversity within L. plantarum was confirmed using Southern profiles, PCR amplification and subsequent sequencing of these PCR products. A 773 bp sequence overlapping the pyrDF genes showed high homology: at least 97% identical in L. plantarum strains (V to IX) and 99.9% identical in hybridisation groups VII and VIII. The same G-T transversion which destroyed the pyrF BglI site was found in 11 strains (hybridisation groups VI, VII and VIII). DNA rearrangements were identified downstream from the pyr genes, by PCR amplification and Southern hybridisation profile analysis in three strains of hybridisation groups VIII and IX, two of which also harboured the G-T transversion. PMID- 11876365 TI - A polyphasic taxonomic study of thermophilic bacilli from shallow, marine vents. AB - Eighty-seven thermophilic, aerobic, spore-forming bacteria were isolated from shallow, marine, thermal vents of the Eolian Islands (Italy) and tested for a broad spectrum of phenotypic characteristics. A numerical taxonomy study was performed on these isolates and 8 thermophilic Bacillus and Geobacillus reference strains by 89 selected features. Results from cluster analysis showed the formation of nine clusters. Most of the isolates (83%) fell into several phenetically well distinguished clusters, loosely related to Geobacillus thermodenitrificans. The remaining isolates grouped together with different reference strains. Eighteen isolates, representative of the different clusters, were selected for subsequent genotypic characterisation, including partial 16S rDNA sequence analysis of 18 strains and almost complete 16S rDNA sequences of 9 strains. Subsequent DNA/DNA reassociation studies and determination of the base composition of DNA identified seven isolates as Geobacillus thermodenitrificans, two isolates as G. thermoleovorans and one isolate as Bacillus pallidus. Four isolates represented two novel species of Bacillus. The remaining four represented novel Geobacillus species, one of which has recently been described as Bacillus vulcani DSMZ 13174 T. PMID- 11876366 TI - The CANON system (Completely Autotrophic Nitrogen-removal Over Nitrite) under ammonium limitation: interaction and competition between three groups of bacteria. AB - The CANON system (Completely Autotrophic Nitrogen Removal Over Nitrite) can potentially remove ammonium from wastewater in a single, oxygen-limited treatment step. The usefulness of CANON as an industrial process will be determined by the ability of the system to recover from major disturbances in feed composition. The CANON process relies on the stable interaction between only two bacterial populations: Nitrosomonas-like aerobic and Planctomycete-like anaerobic ammonium oxidising bacteria. The effect of extended periods of ammonium limitation was investigated at the laboratory scale in two different reactor types (sequencing batch reactor and chemostat). The lower limit of effective and stable nitrogen removal to dinitrogen gas in the CANON system was 0.1 kg N m(-3) day(-1). At this loading rate, 92% of the total nitrogen was removed. After prolonged exposure (> 1 month) to influxes lower than this critical NH4+-influx, a third population of bacteria developed in the system and affected the CANON reaction stoichiometry, resulting in a temporary decrease in nitrogen removal from 92% to 57%. The third group of bacteria were identified by activity tests and qualititative FISH (Fluorescence In Situ Hybridisation) analysis to be nitrite-oxidising Nitrobacter and Nitrospira species. The changes caused by the NH4+-limitation were completely reversible, and the system re-established itself as soon as the ammonium limitation was removed. This study showed that CANON is a robust system for ammonium removal, enduring periods of up to one month of ammonium limitation without irreversible damage. PMID- 11876367 TI - Structure and functional analysis of the microbial community in an aerobic: anaerobic sequencing batch reactor (SBR) with no phosphorus removal. AB - The bacterial community of an aerobic:anaerobic non-P removing SBR biomass fed a mixture of acetate and glucose was analysed using several 16S rRNA based methods. Populations responsible for anaerobic glucose and acetate assimilation were determined with fluorescent in situ hybridization (FISH) in combination with microautoradiography (FISH/MAR). At 'steady state' this community consisted of alpha-Proteobacteria (26%) and gamma-Proteobacteria (14%), mainly appearing as large cocci in tetrads (i.e. typical 'G-Bacteria'). Large numbers of low G+C bacteria (22%), and high G+C Gram-positive bacteria (29%) seen as small cocci in clusters or in sheets were also detected after FISH. DGGE fingerprinting of PCR amplified 16S rDNA fragments and subsequent cloning and sequencing of several of the major bands led to the identification of some of these populations. They included an organism 98% similar in its 16S rRNA sequence to Micropruina glycogenica, and ca. 76% of the high G+C bacteria responded to a probe MIC 184, designed against it. The rest responded to the KSB 531 probe designed against a high G+C clone sequence, sbr-gs28 reported in other similar systems. FISH analyses showed that both these high G+C populations were almost totally dominated by small clustered cocci. Only ca. 2% of cells were beta Proteobacteria. None of the alpha- and gamma-Proteobacterial 'G-bacteria' responded to FISH probes designed for the 'G-Bacteria' Amaricoccus spp. or Defluvicoccus vanus. FISH/MAR revealed that not all the alpha-Proteobacterial 'G Bacteria' could take up acetate or glucose anaerobically. Almost all of the gamma Proteobacterial 'G-Bacteria' assimilated acetate anaerobically but not glucose, the low G+C clustered cocci only took up glucose, whereas the high G+C bacteria including M. glycogenica and the sbr-gs28 clone assimilated both acetate and glucose. All bacteria other than the low G+C small cocci and a few of the alpha Proteobacteria accumulated PHB. The low G+C bacteria showing anaerobic glucose assimilation ability were considered responsible for the lactic acid produced anaerobically by this SBR biomass, and M. glycogenica for its high glycogen content. PMID- 11876369 TI - Viable cytophaga-like bacterium in the 0.2 microm-filtrate seawater. AB - A strain of the Cytophaga-like bacterium (CLB), Nano-1, was isolated from the 0.2 microm-filtrate of natural seawater. Both cellular fatty acid and 16S rDNA sequence analyses indicated that Nano-1 is closely affiliated to the marine gliding CLB genus, Microscilla. Nano-1 was observed to undergo cyclic morphological change typical of the genus Microscilla, and sub-0.2-microm cells were formed in the late stationary phase. The sub-0.2-microm cells were repeatedly revived and subcultured. Formation of the sub-0.2-microm cells seems to be adaptive for oligotrophic growth and starvation survival. PMID- 11876368 TI - The potential of a polyphasic PCR-dGGE approach in evaluating microbial diversity of natural whey cultures for water-buffalo Mozzarella cheese production: bias of culture-dependent and culture-independent analyses. AB - A polyphasic PCR-DGGE approach was used to describe the microbial population occurring in natural whey cultures (NWCs) for water-buffalo Mozzarella cheese production. Total microbial community was assessed without cultivation by analyzing DNA directly extracted from the original samples of NWC. In addition, DNA extracted from bulks of cells formed by harvesting colonies from the serial dilution agar plates of a variety of culture media was used to profile the "cultivable" community. The 16S rDNA V3 region was amplified using DNA from NWC as well as DNA from bulks as templates and the amplicons were separated by DGGE. The microbial entities occurring in NWCs were identified by partial 16S rDNA sequencing of DGGE bands: four lactic acid bacteria (LAB) closest relative of Streptococcus thermophilus, Lactococcus lactis, Lactobacillus delbrueckii and Lactobacillus crispatus were revealed by the analysis of DNA directly extracted from NWC while two other LAB, Lactobacillus fermentum and Enterococcus faecalis, were identified by analyzing DNA from the cultivable community. The developed PCR DGGE analysis of the "cultivable" community showed good potential in evaluating microbial diversity of a dairy environment: it usefully highlighted the bias introduced by selective amplification when compared to the analysis of the total community from NWC and allowed suitability of media and growth conditions to be evaluated. Moreover, it could be used to complete the culture independent study of microbial diversity to give information on concentration ratios among species occurring in a particular environment and can be proposed for rapid identification of dominant microorganisms in alternative to traditional tools. PMID- 11876370 TI - Microbial diversity of cultivatable bacteria associated with the North Sea bryozoan Flustra foliacea. AB - The microbial diversity of cultivatable bacteria associated with the bryozoan species Flustra foliacea from the North Sea was investigated by a molecular approach. Amplified ribosomal RNA restriction analyses (ARDRA) and 16S rDNA partial sequence analysis revealed differences in the composition of cultivatable bacteria populations from single bryozoan colonies collected from two different sampling sites in the North Sea as well from one site taken at different points in time. Whereas gamma-Proteobacteria identified as Shewanella frigidimarina, Pseudoalteromonas ssp. and Psycbrobacter ssp. were predominant on samples of Flustra I (taken near the island of Helgoland), most bacteria isolated from Flustra II, originating from the Steingrund, could be affiliated to Gram-positive taxa. Survey of the bryozoan samples from the latter site in February 2000 led to the detection of a phylogenetically mixed bacterial population, consisting of gamma-, and alpha-Proteobacteria and Gram-positive bacteria with low and high GC content (Flustra III). As these bacteria are among the most widely isolated organisms from the marine environment, it may be concluded that the bryozoan Flustra foliacea accepts colonization of surfaces by bacteria which are common inhabitants of the marine environment and which may have been transferred into this environment from terrestrial sites. PMID- 11876371 TI - Yeasts present during wine fermentation: comparative analysis of conventional plating and PCR-TTGE. AB - Yeasts isolated from must before and during fermentation at a wine cellar of La Mancha region in Spain were characterised using Polymerase Chain Reaction / Restriction Fragments Lengths Polymorphism and Polymerase Chain Reaction / Temporal Temperature Gradient Gel Electrophoresis. S. cerevisiae strains were differentiated using mtDNA restriction analysis. Direct PCR-TTGE was also used to study biodiversity during wine fermentation, and revealed the variations in the population. It was observed that isolation by conventional plating may afford a skewed view of the strains taking part in wine fermentation. PMID- 11876372 TI - Performance of industrial strains of Saccharomyces cerevisae during wine fermentation is affected by manipulation strategies based on sporulation. AB - Genetic manipulation of industrial wine yeast strains has become an essential tool for both the study of the molecular mechanisms underlaying their physiology and the improvement of their fermentative properties. The construction of null mutants for any gene in these usually diploid strains, by using a procedure based on sporulation of a heterozygote lacking one copy of the gene of interest, has been tested as an alternative to the tedious work of sequential disruption of the complete set of copies. Our results indicate that most of the homozygotes resulting from sporulation of wine yeast strains are defective in glucose consumption under microvinification conditions in synthetic must and produce stuck fermentations. These kinds of defects are observed even for strains derived from sporulation of wild type. Alteration of genomic features of wine strains by sporulation is responsible for these defects. PMID- 11876373 TI - Improving diabetes care--can we do more? PMID- 11876374 TI - A window on the current status of diabetes mellitus in Singapore--the Diabcare Singapore 1998 study. AB - The Diabcare-Singapore project was carried out in 22 clinics (general hospitals, GH and primary healthcare centres, PHC) to provide an overview of diabetes management and metabolic control status. Data from 1697 diabetic patients were collected on paper forms and analysed centrally. Type 2 diabetes mellitus patients constituted 91.4% and type I patients constituted 8.1% of population. The proportion of type I patients was greater in GH (18.1%) vs PHC (3.4%). The mean age (+/- SD) was 58.1 +/- 14.4 years and mean duration of diabetes was 10.1 +/- 7.5 years. Mean body mass index (BMI) was 25.1 +/- 4.4 kg/m2 and more than half (53%) of patients were overweight (BMI >25 kg/m2). Mean HbA1c and FBG levels were 8.0 (1.9% and 9.1 +/- 3.1 mmol/l. A total of 51% of patients had HbA1c (1% above the Upper Limits of Normal (ULN). Fasting blood glucose (FBG) was >7.8 mmol/l in 61% of patients. The majority (70%) had satisfactory levels of fasting lipids (triglycerides, total cholesterol and HDL-cholesterol). Only 19.7% practised home blood glucose self-monitoring, while 99% reported receiving some diabetes education. Sixteen percent of patients had abnormal levels of protein (>500 mg/24 h) in the urine, 3% had elevated serum creatinine levels and 36% had microalbuminuria. Retinopathy (12%), cataract (16%) and neuropathy (12%) were commonly reported diabetic complications. The data revealed suboptimal glycaemic control in about half of patients studied. PMID- 11876375 TI - The status of diabetes mellitus in primary care institution and restructured hospitals in Singapore. AB - The Diabcare-Asia Singapore 1998 project was carried out using data from 22 centres collected on paper forms to provide an overview of diabetes management and metabolic control status in 1697 diabetic patients from both primary health care clinic (PHC) (67%) and restructured hospital (RH) (33%) settings. PHC patients were on average older than RH patients (61.3 +/- 11.2 years vs 51.5 +/- 17.7 years), and had a shorter duration of diagnosed diabetes (9.2 +/- 6.8 years vs 12.0 +/- 8.5 years). The mean body mass index (BMI) for PHC patients was 25.5 +/- 4.4 kg/m2 vs 24.5 +/- 4.2 kg/m2 for RH patients. Proportionately more PHC than RH patients were overweight (BMI >25 kg/m2) (49% vs 42%). Patients with type I diabetes constituted 3.5% of PHC vs 18.1% of the RH cohort. HbA1c information was available for 92.5% of RH vs 69% of PHC patients. HbA1c measurements were <1% above ULN in 50% of PHC vs 37% of RH patients, while FBG was >7.8 mmol/l in >61% of all patients. Proteinuria (>500 mg/24 hrs) was reported in 13% of PHC vs 26% of RH patients tested. Microalbuminuria (20-300 mg/l) was noted in 36% of 171 RH patients tested. Oral hypoglycaemic agents were used as sole therapy in 83.5% of PHC vs 43% of RH patients. Eye, feet, renal and severe late complications were more commonly reported by RH than PHC patients. There is a variation in the patient profiles and care between PHC and RH patients. PMID- 11876376 TI - Latent autoimmune diabetes in adults (LADA): a case series. AB - An adult presenting with diabetes is usually assumed to have type 2 diabetes. Since the 1980s, type 2 diabetic subjects who had failed sulphonylurea therapy soon after diagnosis have been thought to be actually slowly progressive type I patients. This diabetes sub-type is currently referred to as latent autoimmune diabetes in adults (LADA). Early recognition of such patients has important clinical implications. To assist local doctors in the recognition of such patients, we performed a retrospective study to profile and highlight distinctive features of thirteen LADA patients. We found that these patients were mostly females with a mean body mass index of 17.2 kg/m2, diagnosed with type 2 diabetes in their fourth decade of life and becoming insulin dependent after a mean of 2.5 years. PMID- 11876377 TI - The glucose challenge test for screening gestational diabetes in pregnant women with no risk factors. AB - AIM OF STUDY: To evaluate the 50 g glucose challenge test as a screening tool for gestational diabetes in pregnant women with no risk factors, to determine the prevalence of gestational diabetes in this population and to determine the perinatal outcomes of pregnancy according to the glucose challenge test. METHODOLOGY: A descriptive prospective study. A total of 146 patients with no risk factors who booked a particular obstetrician and delivered between May 1996 and April 1997 were recruited. Pregnancy outcomes were assessed by the gestation and mode of delivery, neonatal outcomes included birth weights, apgar scores and other neonatal complications. RESULTS: The detected incidence of gestational diabetes was 8.2%. With the threshold plasma glucose level at 7.1 mmol/l, 53 women or 36% needed to undergo the 75 g oral glucose tolerance test and 12 women were found to have gestational diabetes. The diagnostic yield was 22.6%. With 7.8 mmol/l as the threshold value, 28 women or 20% needed the oral glucose tolerance test and eight women with gestational diabetes were detected. The diagnostic yield was 28.6%. Perinatal outcome for these diabetic women who were well controlled during pregnancy was similar to the rest of the women with normal glucose challenge test. CONCLUSIONS: The 50 g glucose challenge test is a useful screening test for diabetes in Singaporean women with no risk factors. A threshold value at 7.8 mmol/l with a smaller number of women requiring the 75 g oral glucose challenge test may be more acceptable. PMID- 11876378 TI - High prevalence of psychiatric morbidity in a medical intensive care unit. AB - This study seeks to determine the prevalence of psychiatric morbidity within a medical intensive care unit, examine its correlation with the various physiological parameters and delineate any clinical predictors for psychiatric morbidity. Seventy-seven patients who gave informed consent were administered the General Health Questionnaire (GHQ), Acute Physiological And Chronic Health Evaluation II (APACHE II) and thyroid function tests were performed. A high prevalence of psychiatric morbidity was found (36.4%). However, no statistically significant association was found between psychiatric morbidity and gender, age, APACHE II scores and thyroid function indices. Nevertheless, it is hoped that the index of suspicion for psychiatric morbidity can be raised in order to optimise the clinical management of patients within this setting. PMID- 11876379 TI - Hypodermoclysis or subcutaneous infusion revisited. AB - AIM OF STUDY: To review the use of hypodermoclysis in a local Hospice. METHOD: A review of all hypodermoclysis carried out over a six-month period was conducted. Special attention was paid to the reason for starting and stopping the drip, duration of the drip, complications, the type and amount of solution infused. RESULTS: Fifty-one (19%) out of 266 patients received hypodermoclysis during their stay. This constituted 5.9% of total patient-days in the study period. Vomiting and drowsiness were the main reasons for the use of drip. The commonest reason for stopping the drip was patient demise. Complications seen were drip site redness (16%), extravasation (15%) and bleeding (2.5%) There was no overt clinical sepsis in any of the patients. CONCLUSION: Hypodermoclysis is an easy and convenient means of providing hydration. The availability of a standard protocol with clearer guidelines on its use will help to reduce procedure-related complications and promote wider adoption of the practice. PMID- 11876380 TI - Acquired immunodeficiency syndrome presenting as childhood non-Hodgkin's lymphoma. AB - Two children with non-Hodgkin's lymphoma (NHL) as the presenting illness of acquired immunodeficiency syndrome (AIDS) are described. There was a delay in diagnosing the underlying AIDS in both cases. In the first case, an 18-month-old boy with stage IV, high-grade,T-cell NHL, the diagnosis of underlying AIDS was suspected only when he developed recurrent and profound opportunistic infection during chemotherapy. The second case, an eight-month-old female infant presented initially with hepatosplenomegaly and thrombocytopenia of undetermined cause. She had progressive abdominal distension and swelling of her right eye one year later due to high grade B-cell NHL. She was later found to be sero-positive for HIV during pre-chemotherapy screening. As the prevalence of HIV infection continues to increase, HIV infection should be considered in the differential diagnoses of childhood hepatosplenomegaly and thrombocytopenia, and as a possible underlying cause of childhood cancer, especially NHL. PMID- 11876381 TI - Gastrointestinal stromal sarcoma--a case report of palliative enteral stenting. AB - We report a case of metastatic gastrointestinal stromal sarcoma (GISS) in a 33 year-old man who subsequently underwent successful palliative endoscopically placed enteral stenting for duodenal stenosis secondary to extrinsic compression. Enteral stenting for palliative relief of malignant gastrointestinal obstruction is recommended for its safety, efficacy and cost-effectiveness. PMID- 11876382 TI - Medical management of cervical pregnancy--a report of two cases. AB - Cervical ectopic pregnancy is a rare life-threatening condition that can be managed conservatively. In this report, the authors describe the use of systemic methotrexate and prostaglandin (sulprostone) in the management of two cases of viable cervical pregnancy. Both cases were successfully treated without the need for surgical intervention. PMID- 11876383 TI - Clinics in diagnostic imaging (67). Hydronephrotic horseshoe kidneys with multiple calculi. AB - Horseshoe kidney is the commonest congenital renal anomaly. Its inherently-poor drainage system renders the patient prone to stone formation. A 56-year-old man with bilateral renal stones in a horseshoe system associated with hydronephrosis is presented. He was treated successfully with bilateral CT-guided percutaneous nephrostomy and percutaneous nephrolithotripsy. Various treatment options, including ESWL, PCNL and open surgery, are discussed. PMID- 11876384 TI - Intraoperative pathological monitorization of surgical margins: a method to reduce recurrences after conservative treatment for breast cancer. AB - INTRODUCTION: Local recurrences after conservative surgical treatment for breast cancer are not uncommon and cause negative influences on the oncological prognosis and quality of life of the patients. Aiming to avoid this problem, we have developed a method of intraoperative pathological monitoring of surgical margins (IPMSM), in order to assure adequacy of resection. MATERIALS AND METHODS: IPMSM is based on radiological. macroscopic, cytological and histological examination of frozen sections of the breast specimens in the operating room during the surgery. We evaluated 98 women with 100 tumors clinical stage I-II breast cancer for whom we planned conservative surgery. The margins were oriented by the surgeon and inked by the pathologist in different colors to retain orientation. RESULTS AND DISCUSSSION: According to the histological or cytological results, immediate re-excision was indicated and performed in 40 (40.8%) cases. In six of these, we had to perform a mastectomy. The indications for additional resections were: insufficient margins in 23 cases, extensive intraductal component in eight, multifocality in four, atypical proliferative lesion at the margin in four and diffuse tumor in one. Permanent histological sections confirmed all intraoperative results. These patients were followed by a median period of 42 months (range 3 to 99 months) and we observed 1% of local recurrence and 5.1% of distant metastasis. We compared this group of patients with a control group represented by 149 cases of breast cancer stages I-II treated by conservative surgery, but not submitted to IPMSM. In the control group, we observed 17 (11.4%) local recurrences and 49 (32.9%) distant metastases after a follow-up period from 14 to 213 months (median of 126 months). CONCLUSION: The IPMSM proved to be a safe and accurate method to prevent additional surgery for insufficient margins and to reduce the recurrence rate. PMID- 11876385 TI - Both TPA and SCC-Ag levels are prognostic even in high-risk stage Ib-IIa cervical carcinoma as determined by a stratification analysis. AB - OBJECTIVE: To determine the prognostic values of tissue polypeptide antigen (TPA), squamous cell carcinoma antigen (SCC-Ag), and carcinoembryonic antigen (CEA) in the sera of cervical carcinoma patients, especially in those with a poor prognosis. METHODS: In this retrospective study, the preoperative serum SCC-Ag, TPA, and CEA were analyzed in 779 patients with cervical squamous cell carcinoma of stage Ib-IIa who received radical hysterectomy and pelvic lymph node dissection (RAH-PLND) between 1984 and 1994. RESULTS: Due to poor predictive value and poor correlation between serum CEA and clinico-pathological factors, CEA was abandoned in this study. Elevated TPA and SCC-Ag levels, pelvic lymph node metastasis (PLNM), lymphvascular space involvement (LVSI) and deep stromal invasion (DSI) were associated with poor survival time by univariate analysis. The correlation study showed that elevated serum TPA was significantly related to PLNM, LVSI, and DSI (p = 0.004, 0.008, and 0.021, respectively), and SCC-Ag was related to PLNM and bulky tumor size (p = 0.001 and 0.02, respectively). In the multivariate analysis, only PLNM and LVSI remained independently significant indicating poor survival. Further stratification studies by PLNM and LVSI showed that elevated TPA levels could even indicate higher recurrence rates in patients with PLNM (p = 0.045), as well as SCC-Ag in patients with LVSI (p = 0.038). CONCLUSIONS: The results suggest that both elevated TPA and SCC-Ag levels depicting poor prognosis in stage Ib-IIa cervical SCC, especially indicates a group of high-risk patients who may need more aggressive therapy. PMID- 11876386 TI - Primary breast carcinoma of the vulva: case report and review of literature. AB - The occurrence of ectopic breast tissue within the vulva is uncommon and the development of breast carcinoma within vulvar ectopic breast tissue is very rare. To date, only 12 cases of primary vulvar breast carcinoma have previously been reported in the English literature. This paper presents the 13th reported case of primary breast carcinoma of the vulva. The patient presented with a vulvar ulcerated lump and the diagnosis was based on a morphologic pattern consistent with breast carcinoma and the presence of estrogen and progesterone receptors. Primary surgery consisted of radical vulvectomy and bilateral groin dissection. The groin lymph nodes were involved bilaterally. Adjuvant therapy consisted of systemic chemotherapy (4 cycles of adriamycin and cyclophosphamide followed by 4 cycles of paclitaxel) and pelvic radiotherapy. Oral tamoxifen 20 mg/day was started for the next five years. It is concluded that the management of primary breast carcinoma of the vulva should be modeled after that for primary carcinoma of the orthotopic breast with primary surgery followed by systemic chemotherapy and pelvic radiotherapy. Chemotherapy should be similar to that employed for breast carcinoma. Tamoxifen should be prescribed for patients whose tumors contain estrogen receptors. PMID- 11876387 TI - Aggressive endometrial carcinoma in a breast cancer patient treated with tamoxifen with normal transvaginal ultrasonography. Case report. AB - Since tamoxifen therapy can induce endometrial disorders, surveillance schemes of women taking tamoxifen have been recommended. Transvaginal ultrasonography is a very sensitive test and therefore is often performed as a first-line screening test. We described a very atypical case of a high stage, high grade endometrial cancer associated with tamoxifen in a 64-year-old woman with a past history of breast cancer. This women was assessed yearly by ultrasonography and Pap smear. The cancer developed on a very thin endometrium and transvaginal ultrasonography failed to detect it. The patient remained asymptomatic up to the diagnosis. Normal endometrial cells in the Pap smear test were the only signs associated with this cancer. Surveillance strategies and significance of endometrial cells on the Pap smear are reviewed. In conclusion, TVUS can fail to detect cancers if the endometrial lining is not enlarged. In case of normal endometrial cells in the Pap smear, a careful evaluation should be performed. PMID- 11876388 TI - Color Doppler ultrasonography in the differentiation of uterine sarcomas from uterine leiomyomas. AB - OBJECTIVE: The aim of this study was to investigate uterine vascularity in cases of uterine leiomyomas and uterine sarcomas, as well as to determine the efficiency of uterine blood flow analysis in differentiating between them. MATERIALS AND METHODS: Transvaginal color and pulsed Doppler findings obtained from 117 patients with histologically proven uterine leiomyoma and 12 with uterine sarcoma were retrospectively assessed. RESULTS: The mean intratumoral resistance index (RI) and pulsatility index (PI) were significantly lower and the intratumoral peak systolic velocity (PSV) was significantly higher in patients with sarcomas than in patients with uterine leiomyomas. Marked reduction of RI and PI and increased PSV could be found in 14 of the leiomyoma cases which showed large size and/or necrotic, degenerative and inflammatory changes. When a cut-off value of 0.5 for the RI was considered, the detection rate for uterine sarcoma was 67% and the false-positive rate was 11.8%. CONCLUSION: These results suggest that the intratumoral RI detected by color and pulsed Doppler ultrasonography in themselves could be poor for the preoperative differential diagnosis of uterine sarcoma. PMID- 11876389 TI - Impact of screening on breast cancer detection. Retrospective comparative study of two periods ten years apart. AB - OBJECTIVE: The aim of this study was to evaluate changes in the mode of discovery of breast cancer in the last 15 years. We compared two periods separated by a 10 year interval, during which a mass mammographic screening programme was established in our department. MATERIALS AND METHOD: We made a retrospective comparison of the records of female patients with breast cancer diagnosed in our hospital over the period 1986-1989 (first period) and 1997-1999 (second period). The mass screening programme for breast cancer began in 1995. RESULTS: We collected 372 patients in the first period and 341 in the second. We found a significant change in the mode of the discovery of breast cancer between the two periods: 80.2% versus 51.9%, respectively, of the cases of breast cancer were discovered by breast self-examination, 10.2% versus 13.7% were discovered by a physician, and 4.8% versus 29.1% were discovered by routine mammography as part of an individual or mass screening programme. The mean size of the tumours decreased significantly (2.6 cm versus 2.3 cm: p = 0.019), and the number of tumours with initial metastases or lymph node involvement decreased, almost attaining the level of significance (p = 0.06). It is difficult to compare the survival and disease-free survival curves because of the short follow-up in the second period (median follow-up = 10 months). However, a marked difference appears to be developing (p < 0.0001): patients diagnosed by mammography are showing better survival and disease-free survival compared with the others. DISCUSSION: We observed that more widespread use of mammography screening for breast cancer led to smaller tumours being discovered during the second period, with less lymph node involvement and less initial metastasis. Breast cancer screening is one of the most intensively evaluated health care practices with eight completed randomized trials yet its net benefit has remained controversial. It has been shown that, at least for patients aged 50 to 70, properly organized mass screening for breast cancer led to a reduction in mortality rate. However, individual breast self-exam, physician and mammographic screening can interfere with assessment of mass screening programmes in terms of individual benefit. In addition, introducing a mass screening programme may induce opportunistic screening in non-invited age groups and influence health behaviour in the target and non target populations. A retrospective study was performed to evaluate the mode of discovery, the diagnostic presentation, and prognostic factors in breast cancer in a French department before and after initiation of a mass-screening programme (MSP). PMID- 11876390 TI - Correlation between atypical colposcopy findings and detection of human papillomavirus (HPV) infection of the uterine cervix. AB - Reports on the detection of genome human papillomaviruses (HPV) in genital neoplasia differ to a great extent either in the overall prevalence or in the frequency of certain types. The aim of the study was to determine the correlation between the HPV infection and the occurrence of premalignant and malignant diseases of the uterine cervix and to investigate the ratio between clinical features and infection findings starting from the assumption that infection by human papillomaviruses is a key factor in the occurrence of premalignant and malignant disease of the uterine cervix. The investigation was carried out on 48 patients who formed the study group (Group I). Based on suspicious colposcopy findings, a Papanicolaou (Pap) smear and biopsy were performed and a histopathological analysis of the sample was carried out. A cervical smear was done on all the patients for HPV detection and typing. The patients in whom HPV infection was not found formed a control group (C Group). In spite of certain divergences it has not been proved that the ratio between colposcopy findings and HPV type has any statistical importance (chi2 = 3.305; p > 0.05). The distribution of Pap smear results did not shown a significant difference with respect to HPV type (chi2 = 0.105; p > 0.05). When the data are analyzed the diagnosis of low-grade squamous intraepithelial lesions (LGSIL) is evident in 20% of the HPV cases whereas it is significantly lower with respect to the group where HPV was not detected (42.5%). Histopathological (HP) findings of a high grade squamous intraepithelial lesions (HGSIL) in both groups are diagnosed in approximately the same percentage while 6.7% of cancer in situ was registered in the group of HPV positive patients. Based on this it can be concluded that if a diagnosis of LGSIL or HGSIL in particular has been made on the basis of HP findings there is a great probability that the infection was due to one or more joined types of human papillomaviruses. PMID- 11876391 TI - Abnormal "low grade" transformation zone: current diagnostic gold standard. AB - The aim of this work was to examine different methods of investigation in the diagnosis of the abnormal "low grade" transformation zone of the portio. Over a period of one year 41 patients subjected to colposcopic examination underwent exo endocervical sampling for oncologic evaluation and for detection of viral and bacterial infections (HPV, HSV, adenovirus, mycoplasmas and chlamydia trachomatis), as well as portio biopsy. A 65.8% correlation was found between cytology and the HPV-DNA test results, while histology and the presence of the HPV virus agreed in 51.4% of cases. In those cases in which minimal histological alterations were found (koilocytosis) a high percentage of HPV negativity was found. In discordant negative cytologic tests that were however positive for HPV by PCR, the genotypes identified were always 6 and 11. PMID- 11876392 TI - Strategies for the modification of risk factors in gynecological cancers. AB - Strategies to modify risk for female or gynecological cancers will vary with our knowledge of the epidemiology, etiology, and specific molecular mechanisms for each individual cancer. In general, cancer preventive strategies have been divided into primary and secondary prevention with primary prevention directed toward the causative factors for a disease. Secondary prevention is classically used in cervical cancer cytology screening programs and is essentially an attempt to identify individuals in a population with preclinical phases of the disease where intervention will impact mortality the most. A vast literature has evolved regarding the epidemiology of most of the common cancers in women. While the specific molecular mechanisms are not completely understood at this time knowledge of contributing factors for many of these tumors is well known. The association of cigarette smoking with lung cancer has been well established and the increasing rates of lung cancer, particularly in women, are directly linked to the increasing number of female smokers in the population. Indeed in many western countries lung cancer deaths have overtaken breast cancer as the most common cause of death from malignant disease in women. Excessive sun exposure without adequate skin protection is another lifestyle activity that is related to the high incidence of skin cancer in certain areas. Epidemiologically, cervical cancer has been studied extensively with the current data indicating a causal role of exposure to human papillomavirus (HPV), particularly at an early age in this disease. Hereditary breast and ovarian cancer syndromes are well understood and as more information on human genomics becomes available a clear understanding of the underlying molecular mechanisms of these diseases will be possible and hopefully will result in effective strategies for their control. Unfortunately, in spite of the vast knowledge that is available regarding risk factors for many of these malignancies we have been unable to influence effective lifestyle changes that could substantially reduce the risk of these malignancies in our population. Increased efforts in education, research, and commitment--both financial and educational--are required by governments and other social organizations. PMID- 11876393 TI - The effects of tamoxifen on rat skin. AB - BACKGROUND: Tamoxifen (Tx) is used mostly in the treatment of breast and gynecological cancers. It is also widely used in the treatment of different dermatological disorders. However, its effects on skin have not been investigated previously. OBJECTIVE: To investigate the effects of Tx administration on rat skin. METHODS: Forty Spraque-Dawley female newborn rats were separated into two control groups and two experimental groups (n 10). One day after birth, the control groups of newborn rats were given 0.02 ml saline subcutaneously (s.c.) daily whereas experimental litters were treated with 100 microg Tx citrate in 0.02 ml saline s.c. daily for five days. The first control group and experimental group of rats were anesthetized at 21 days whereas the second control group and experimental group of rats were anesthetized on the 28th day. Histopathological assessments were made and compared with the control groups. RESULTS: Abnormal hair follicles were observed in both experimental groups of rats. Epidermal atrophy together with increased dermal fibrosis was more prominent in the first experimental group. Dermal fibrosis and lymphohistiocytic inflammatory cell infiltration were found to be prominent around the hair follicles in the second experimental group. CONCLUSION: Considerable harmful effects of Tx administration were observed on rat skin. PMID- 11876394 TI - Comparison of two procedures for sentinel lymph node detection in patients with endometrial cancer: a pilot study. AB - OBJECTIVE: The purpose of this study was to assess the feasibility and contribution of two intraoperative procedures of lymphatic mapping and sentinel node detection using a blue dye in surgically-staged patients with early stage endometrial cancer. METHODS AND MATERIALS: In 25 cases of endometrial cancer, patent blue-V was injected into the subserosal myometrium (13 cases, SM group) or cervico-subserosal myometrium (12 cases, CSM group) during a surgical staging procedure. Laparoscopically-assisted vaginal hysterectomy and pelvic lymphadenectomy were completed successfully in 23 women out of 24 laparoscopically-staged patients (95.8%). One patient with FIGO stage IIa was indicated for a radical abdominal surgery. RESULTS: A deposition of the blue dye was found in at least one pelvic lymph node (LN) in eight out of 13 cases (61.5%) in the SM group compared with ten out of 12 cases (83.3%) in the CSM group (p = 0.378). The mean number of dye-colored LN (DCLN) was 1.15 (SM group) and 2.5 (CSM group), respectively (p = 0.05). The rate of DCLN/LN was 15/188 (SM group) versus 30/190. respectively (p = 0.03). An uptake of the blue bye was observed in a total of 45 out of 388 LN. CONCLUSION: An intraoperative combination of cervico subserosal myometrium application of the blue dye allows successful detection (83.3%) of sentinel LN in patients with endometrial cancer. Comparing SM and CSM groups the statistical significant difference was found in the DCLN/LN rate and mean number of sentinel lymph nodes (p = 0.03, p = 0.05, respectively). Clinical validity of this surgical procedure must be assessed prospectively. PMID- 11876395 TI - Evaluation of morbidity after external radiotherapy and intracavitary brachytherapy in 771 patients with carcinoma of the uterine cervix or endometrium. AB - PURPOSE: The aim of the present study was to evaluate early and late radiation morbidity and to assess the factors influencing morbidity in patients with cervical or endometrial cancer treated by a combination of external radiotherapy (ERT) and intracavitary brachytherapy (IBRT). MATERIALS AND METHODS: Early and late radiation morbidity were evaluated retrospectively using RTOG/EORTC criteria and Franco-Italian glossary in 771 patients treated between November 1992 and December 1999. RESULTS: Four hundred and seven patients (52.8%) had endometrial carcinoma and 364 (47.2%) had carcinoma of the cervix. One hundred and fifty-four patients with cervical carcinoma were inoperable. In patients with endometrial carcinoma total doses at the vagina, bladder and rectum were 60.36 Gy, 56.2 Gy and 55.6 Gy respectively. Biologically equivalent doses (BED) for the same points were 79.35, 68.63 and 67.37, respectively for early effects and 123.67, 97.65 and 94.85, respectively for late effects. One hundred and sixty-nine patients (41.5%) developed acute morbidity, grade I and II bladder morbidity being the most common type and 85 patients (20.9%) developed late morbidity, grade I and II vaginal morbidity being the most common type. No grade IV morbidity was recorded. Total doses at the vagina, bladder and rectum in operated cervix cancer patients were 60.51 Gy, 56.53 Gy and 55.67 Gy, respectively. BED for the same points were 79.77, 69.36 and 67.52, respectively for early effects and 124.74, 99.3 and 95.17, respectively for late effects. Eighty patients (38.1%) developed early morbidity. Grade I and II bladder morbidity was the most common type. Sixty-five patients (30.9%) developed late morbidity, vaginal morbidity being the most common type. Total doses at the vagina, bladder and rectum in inoperable patients were 70.92 Gy, 66.71 Gy and 62.38 Gy, respectively. BED for the same points were 97.43, 89.64 and 81.63, respectively for early effects and 159.3, 143.16 and 126.56, respectively for late effects. Sixty patients (39%) developed acute morbidity which was grade I or II bladder morbidity in 95%. Ninety-five patients (61.7%) developed late morbidity which was grade I-III vaginal morbidity in 94%. CONCLUSION: Patients with cervical or endometrial cancer can be treated safely by a combination of ERT and IBRT. However the patients should be assessed before, during and after treatment and at every period of follow-up using a standard and well-defined system in order to define and predict the morbidity rate. PMID- 11876397 TI - Intraoperative placement of a self-retaining Foley catheter for continuous drainage of malignant ascites. AB - Malignant ascites in advanced cancer is usually treated by repeated paracentesis, causing both discomfort and inconvenience to patients in the terminal stages of disease. We present a case of advanced ovarian carcinoma in which intraoperative placement of a Foley's self-retaining catheter into the peritoneal cavity was used to facilitate long-term continuous drainage of malignant ascites. This is a simple, convenient and cost-effective method which decreases the need for repeated hospital admissions. The aim complication might be peritonitis, but with proper care of the device and the use of antibiotics, this was not seen in our patient. PMID- 11876396 TI - Mechanism of action of Tripterygium Wilfordii polyglycoside on experimental endometriosis. AB - PURPOSE OF INVESTIGATION: This study was designed to examine the therapeutic effectiveness and mechanism of action of Tripterygium Wilfordii polyglycoside (TWP) in the treatment of endometriosis. METHODS: An experimental endometriosis model was developed using New Zealand White rabbits where endometrial tissue was autotransplanted into the peritoneum. Six weeks after transplantation, a total of 22 rabbits were randomly placed into two groups: Group I (n=17) was treated with TWP (10 mg/kg/day) and Group 2 (n=5) served as the water-fed control for three successive months. The volume of endometrial implants was measured before and after administration of TWP and water. Immune and endocrine systems were investigated in the normal phase, six weeks after induction of endometriosis, and three months after TWP treatment and water administration. RESULTS: After treatment with TWP, the average volume of endometrial implants significantly decreased (p < 0.0001), and the antiendometrial antibody (EmAb) level decreased (p < 0.05) to near normal levels, but it did not decrease in the untreated controls. Serum FSH and LH levels also decreased after TWP treatment. Furthermore, electron microscopic examination of the pituitary ultrastructure revealed morphological changes in gonadotropic cells (G-cell) after treatment with TWP, and changes gradually disappeared four weeks after withdrawal of TWP. CONCLUSION: This study indicates that TWP has both hormonal and immune system action that is effective as a medical treatment for experimental endometriosis by modulating both reproductive endocrine functions and immunosuppression that results in remission of the disease. PMID- 11876398 TI - Immunohistochemical investigation of p-53, C-NEU and EGFR expression in HPV related epidermoid endometrial carcinoma. AB - Epidermoid carcinoma (PSCC) of the endometrium is a rare form of endometrial cancer that constitutes about 0.1% of all malignant epithelial tumors of the uterus. The diagnosis of PSCC is based on strict criteria and is made in the absence of a glandular component of the tumor. Squamous cell carcinoma of the endometrium should enter the differential diagnosis in postmenopausal patients in the presence of atypical squamous cells in the uterine curettage, while the cervical biopsies are negative for malignancy. The presence of HPV should be investigated as well, so that its pathogenetic relation is clarified. While no significant relation was found to p-53, C-NEU and EGFR expression this investigation must be continued because. HPV may interact with tumor suppressor genes. PMID- 11876399 TI - Cavernous hemangioma of the uterus (a case report). AB - Cavernous hemangioma of the uterus is an extremely rare lesion. We report a postmenopausal patient with abnormal uterine bleeding due to hemangioma and simple endometrial hyperplasia. PMID- 11876400 TI - Detection of high-risk HPV (16, 18, 33) in situ cancer of the cervix by PCR technique. AB - OBJECTIVE: The purpose of this study was to collect data about the incidence of high-risk HPV (16, 18, 33) types in in situ cervical cancers, and to evaluate the reliability of the morphological signs of HPV infection by comparing the presence of these signs to the PCR-proven HPV virus infection. METHODS: Fifty patients who underwent conisation at the Department of Obstetrics and Gynecology of Semmelweis University, Budapest, Hungary because of in situ cervical cancer were examined retrospectively for the presence of HPV infection by the PCR technique. The direct and indirect morphological signs of HPV infection identified in the histological and cytological samples were compared to the actual results of virus DNA amplification by PCR in the identical histological sections. The evaluation of the cytological smears and the histological sections was accomplished independently by two different pathologists. RESULTS: E6 open reading frame of HPV 16, 18 or 33 was detected by PCR in 56% (28 cases) of the histological sections of the 50 examined patients with in situ cancer. In 92% (26 patients) of the 28 HPV positive patients one HPV type was detected, while in one of the remaining two cases two HPV types (16/33), or all three types could be detected. The direct morphological signs for HPV infection proved to be 75% sensitive and 50% specific when compared to the results of PCR. Their predictive value for HPV infection was 65%. For the indirect HPV signs the sensitivity was 64% and specificity 31%. The predictive value, prognosticating the presence of HPV 16, 18, 33 infection was 54% in the same sections. Using significance analysis no significant relationship (p = 0.7728) could be detected between the positivity of indirect signs and the presence of HPV 16, 18, 33 infection, while in case of direct signs the relationship was almost significant (p = 0.0675). The joint testing of the direct and indirect signs did not improve the results (p = 0.1338). During the review of the cytological smears the specificity of the cytology in predicting true HPV infections was found to be 68% and sensitivity was 20%. The predictive value was only 50%. A significance analysis was not accomplished by this diagnostic method because of the missing data (see text). CONCLUSION: The method of Nawa et al. seems to be a reliable approach for the detection of HPV DNA in paraffin-embedded material. The three main types of HPV (16, 18, 33) are probably represented in lower percentages in CIN III in Hungary, but a larger survey is needed to obtain reliable data. The direct and indirect morphological signs of HPV infection failed to show a significant relationship with the PCR proven presence of HPV 16, 18, 33. PMID- 11876401 TI - Expressions of p53, proliferating cell nuclear antigen, and Ki-67 in gestational trophoblastic diseases. AB - OBJECTIVE: This study was done to determine whether the expressions of p53, PCNA, and Ki-67 could differentiate spontaneous abortions with hydropic changes from gestational trophoblastic diseases. MATERIALS AND METHODS: Twenty partial hydatidiform moles, 21 complete hydatidiform moles, nine invasive hydatidiform moles, three choriocarcinomas and 19 first trimester hydropic spontaneous abortions were evaluated by means of immunohistochemical methods with antibodies to p53, PCNA, and Ki-67 in this study. RESULTS: The Ki-67, PCNA, and p53 immunoreactivity was significantly higher in the gestational trophoblastic disease group than in the spontaneous abortion group with hydropic changes. None of the three parameters provided reliable discrimination among gestational trophoblastic disease subgroups. CONCLUSION: Our findings suggest that expressions of Ki-67, proliferating cell nuclear antigen and p53 can be used to differentiate between spontaneous abortion with hydropic changes and gestational trophoblastic disease when all three markers are used together. PMID- 11876403 TI - Lymphangioma circumscriptum of the vulva. PMID- 11876402 TI - Unthreatened late pregnancy with a huge mucinous cyst adenoma of the left ovary: report of an unusual case. AB - Benign cystadenomas or cystic teratomas are most frequently diagnosed in pregnancy. In the latter half of pregnancy ovarian tumors are particularly difficult to diagnose. In this report we present a case of a huge mucinous cyst adenoma of the ovary diagnosed in the 26th week of pregnancy. To our knowledge this is the first report of a case of unthreatened late pregnancy with a huge mucinous cyst adenoma of the ovary. PMID- 11876404 TI - Genetic tools for cyanobacteria. AB - Cyanobacteria are oxygenic photosynthetic bacteria that have been used increasingly to study diverse biological processes, including photosynthesis and its regulation; cell differentiation and N2 fixation; metabolism of nitrogen, carbon, and hydrogen; resistance to environmental stresses; and molecular evolution. Many vectors and other genetic tools have been developed for unicellular and filamentous strains of cyanobacteria. Transformation, electroporation, and conjugation are used for gene transfer. Diverse methods of mutagenesis allow the isolation of many sought-for kinds of mutants, including site-directed mutants of specific genes. Reporter genes permit measurement of the level of transcription of particular genes, and assays of transcription within individual colonies or within individual cells in a filament. Complete genomic sequences have been obtained for the unicellular cyanobacterium, Synechocystis sp. strain PCC 6803 and the filamentous, heterocyst-forming cyanobacterium, Anabaena sp. strain PCC 7120. Genomic sequence projects are under way for Nostoc punctiforme strain PCC 73102 (ATCC 29133) and strains of the unicellular genera, Synechococcus, Prochlorococcus, and Gloeobacter. Genomic sequence data provide the opportunity for global monitoring of changes in genetic expression at transcriptional and translational levels in response to variations in environmental conditions. The availability of genomic sequences accelerates the identification, study, modification and comparison of cyanobacterial genes, and facilitates analysis of evolutionary relationships, including the relationship of chloroplasts to ancient cyanobacteria. The many available genetic tools enhance the opportunities for possible biotechnological applications of cyanobacteria. PMID- 11876405 TI - The elusive roles of bacterial glutathione S-transferases: new lessons from genomes. AB - Glutathione S-transferases constitute a large family of enzymes which catalyze the addition of glutathione to endogenous or xenobiotic, often toxic electrophilic chemicals. Eukaryotic glutathione S-transferases usually promote the inactivation, degradation or excretion of a wide range of compounds by formation of the corresponding glutathione conjugates. In bacteria, by contrast, the few glutathione S-transferases for which substrates are known, such as dichloromethane dehalogenase, 1,2-dichloroepoxyethane epoxidase and tetrachlorohydroquinone reductase, are catabolic enzymes with an essential role for growth on recalcitrant chemicals. Glutathione S-transferase genes have also been found in bacterial operons and gene clusters involved in the degradation of aromatic compounds. Information from bacterial genome sequencing projects now suggests that glutathione S-transferases are present in large numbers in proteobacteria. In particular, the genomes of three Pseudomonas species each include at least ten different glutathione S-transferase genes. Several of the corresponding proteins define new classes of the glutathione S-transferase family and may also have novel functions that remain to be elucidated. PMID- 11876406 TI - 13C-NMR analysis of glucose metabolism during citric acid production by Aspergillus niger. AB - The effect of glucose concentration on glycolytic metabolism under conditions of citric acid accumulation by Aspergillus niger was studied with 13C-labelled glucose. The results show that during cultivation at high glucose (14%, w/v), most of the label in citric acid is in C-2/C-4, and is thus due to the pyruvate carboxylase reaction. However, a significant portion is also present in C-1/C-5, whose origin is less clear but most likely due to reconsumption of glycerol and erythritol. Formation of trehalose and mannitol is high during the early phase of fermentation and declines thereafter. The early fermentation phase is further characterized by a high rate of anaplerosis from oxaloacetate to pyruvate, which also decreases with time. At low glucose concentrations (2%, w/v), which lead to a significantly reduced citric acid yield and formation rate, labelling of citrate in C-2/C-4 is decreased and C-l/C-5 labelling increased. Growth on 2% glucose is also characterized by an appreciable scrambling of mannitol and considerable backflux from mannitol to trehalose (indicating tight glycolytic control at the fructose-6-phosphate step) and an increased anaplerotic formation of pyruvate from oxaloacetate. These data indicate that cultivation on high sugar concentrations shifts control of glycolysis from fructose-6-phosphate to the glyceraldehyde-3-phosphate dehydrogenase step. PMID- 11876407 TI - Polygalacturonase production by Aspergillus awamori on wheat in solid-state fermentation. AB - The production of exo-polygalacturonase (exo-PG) and endo-PG by Aspergillus awamori grown on wheat in solid-state fermentation was studied. Endo- and exo-PG activities were detected after 24 h of inoculation. Glucose released from starch hydrolysis acted as a catabolite repressor for the exo-PG enzyme. In contrast, endo-PG production was not affected by glucose repression. When milled grains were used, the particle-size distribution and the chemical composition of the medium influenced the rate of micro-organism growth and therefore the trend followed by endo- and exo-PG production. However, these two parameters did not affect the maximum production of exo-PG and endo-PG. For one of the milled samples, three different moisture contents were used (50, 55, 60%). Moisture contents of 60% provide a higher yield of pectinases by A. awamori. PMID- 11876408 TI - Production and biotransformation of 6-pentyl-alpha-pyrone by Trichoderma harzianum in two-phase culture systems. AB - The final concentration of 6-pentyl-a-pyrone (6PP) produced in cultures of Trichoderma spp. is limited by the fact that inhibition of biomass growth occurs at 6PP concentrations as low as 100 mg/l. The aim of this work was to evaluate liquid-liquid extractive fermentation systems as an alternative to overcome the toxicity problems and to increase the production of 6PP by this fungus. Two alkanes (n-decane and n-hexadecane) and two dicarboxylic esters (dibutyl phthalate and dioctyl phthalate) were evaluated in shake flask cultures. The highest 6PP production (173 ppm) was achieved when n-hexadecane was used, being 3.5-fold the maximum 6PP concentration of a culture without the solvent. Cultivation of Trichoderma harzianum in a 10-1 bioreactor with n-hexadecane yielded 6PP production ninefold higher than that from control cultures. However, 6PP production in the bioreactor (83 ppm) was lower than in shake flasks. Differences in the power drawn to the fluid at each scale could account for such behavior. Even in the presence of the solvent, 6PP content decreased after reaching its maximal concentration. PMID- 11876409 TI - Effect of shear stress on cultivation of Bacillus thuringiensis for thuringiensin production. AB - Cultivation of Bacillus thuringiensis for thuringiensin production is a mixed growth-associated system. Cultivation conditions should be different during the cell growth stage and production stage. In this study, agitation speed and aeration rate were varied during the exponential growth phase and stationary phase in order to investigate the effect of shear stress via agitation on cultivation of B. thuringiensis for thuringiensin production. It was found that shear stress had a significant effect on thuringiensin production during the stationary phase. By decreasing the agitation speed during the stationary phase, product formation was increased up to 43%. PMID- 11876410 TI - Effects of genotypes of maitake (Grifola frondosa) on biological efficiency, quality and crop cycle time. AB - Twenty-three genotypes of maitake (Grifola frondosa) from Asia, North America and Europe were compared for biological efficiency (BE), quality and crop cycle time. Significant differences among lines were found for BE and crop cycle time when mushrooms were produced on nutrient-supplemented (15% millet, 10% wheat bran and 0.2% gypsum) oak sawdust substrate. Four isolates (WC828, M036, M037, and M040) were found to have the most consistent and highest BEs (38.5%, 39.5%, 35.8%, and 38.9%, respectively) and quality ratings (1.2, 1.3, 1.4, and 1.2, respectively, where I is highest quality and 4 is lowest quality). A commercial line (M039) used in China had the shortest crop cycle time (8 weeks). Lines deemed to have the best commercial potential were of Asian origin. PMID- 11876411 TI - A rapid fluorescence-based assay for detecting soluble methane monooxygenase. AB - A fluorescence-based assay was developed to estimate soluble methane monooxygenase (sMMO) activity in solution. Whole cells of Methylosinus trichosporium OB3b expressing sMMO were used to oxidize various compounds to screen for fluorescent products. Of the 12 compounds tested, only coumarin yielded a fluorescent product. The UV absorbance spectrum of the product matches that of 7-hydroxycoumarin, and this identification was confirmed by 13C-NMR spectroscopy. The dependence of the fluorescent reaction on sMMO activity was investigated by pre-incubation with acetylene, a known inhibitor of sMMO activity. Apparent kinetic parameters for whole cells were determined to be Km(app)=262 microM and Vmax(app)=821 nmol 7-hydroxycoumarin min(-1) mg protein( 1). The rate of coumarin oxidation by sMMO correlates well with those of trichloroethylene degradation and naphthalene oxidation. Advantages of the fluorescence-based coumarin oxidation assay over the naphthalene oxidation assay include a more stable product, direct detection of the product without additional reagents, and greater speed and convenience. PMID- 11876412 TI - Secretion of human interferon alpha 2b by Streptomyces lividans. AB - Biologically active human interferon alpha 2b (HuIFNalpha-2b) was secreted into the culture medium by Streptomyces lividans transformed with recombinant plasmids coding for HuIFNalpha-2b fused to the Streptomyces exfoliatus M11 lipase A signal sequence. Levels were low, 15 or 100 ng/ml, depending on the plasmid used. Neither processed nor unprocessed HuIFNalpha-2b was detected in cell lysates of the transformants secreting the recombinant product. However, the secreted recombinant product was found to partially degrade when cultures reached the stationary phase by the action of an, as yet, unidentified mycelium-associated factor. Experimental evidence suggests that the degrading factor is related to mycelium-associated proteolytic activity. PMID- 11876414 TI - The development of low temperature inactive (Lti) baker's yeast. AB - The construction of a novel baker's yeast variety via traditional genetic techniques is described. The phenotype was designated "Lti" ("Low temperature inactive"). Lti mutations with the desired characteristics within a genetically well-defined haploid laboratory strain of Saccharomyces cerevisiae were isolated, and two different approaches were taken to obtain baker's yeast strains, which exhibit reduced fermenting activity at refrigeration temperatures. In a first approach, a chosen Lti strain carrying mutation lti9 was combined with other laboratory strains carrying defined MAL alleles. In a second approach, the same lti mutation was introduced in the genetic background of polyploid commercial baker's yeast strains that harbor important "industrial" properties. Lti strains arising from both approaches were characterized with specifically developed screening procedures. Strains of the "academic" Lti strain family displayed between 85% and 92% of the biomass yield of a commercial reference strain, whereas strains of the "industrial" Lti strain family showed a variation between 60% and 115%. Lti strains from both families varied strongly among each other in their activity in model doughs: at 8 degrees C they displayed activities between 5% and 30%, and at 30 degrees C between 40% and 113% of a commercial reference baker's yeast strain. PMID- 11876415 TI - A novel methodology employing Corynebacterium glutamicum genome information to generate a new L-lysine-producing mutant. AB - Classical whole-cell mutagenesis has achieved great success in development of many industrial fermentation strains, but has the serious disadvantage of accumulation of uncharacterized secondary mutations that are detrimental to their performance. In the post-genomic era, a novel methodology which avoids this drawback presents itself. This "genome-based strain reconstruction" involves identifying mutations by comparative genomic analysis, defining mutations beneficial for production, and assembling them in a single wild-type background. Described herein is an initial challenge involving reconstruction of classically derived L-lysine-producing Corynebacterium glutamicum. Comparative genomic analysis for the relevant terminal pathways, the efflux step, and the anaplerotic reactions between the wild-type and production strains identified a Val-59-->Ala mutation in the homoserine dehydrogenase gene (hom), a Thr-311-->Ile mutation in the aspartokinase gene (lysC), and a Pro-458-->Ser mutation in the pyruvate carboxylase gene (pyc). Introduction of the hom and lysC mutations into the wild type strain by allelic replacement resulted in accumulation of 8 g and 55 g of L lysine/l, respectively, indicating that both these specific mutations are relevant to production. The two mutations were then reconstituted in the wild type genome, which led to a synergistic effect on production (75 g/l). Further introduction of the pyc mutation resulted in an additional contribution and accumulation of 80 g/l after only 27 h. This high-speed fermentation achieved the highest productivity (3.0 g l(-1) h(-1)) so far reported for microbes producing L lysine in fed-batch fermentation. PMID- 11876416 TI - Acid-base enrichment enhances anaerobic hydrogen production process. AB - This study offers a novel and quick enrichment technology that can be used as a preliminary method to obtain a hydrogen-producing species from the biological sludge produced by wastewater treatment. The influences of acid-base enrichment (by sludge pH adjustment) on the anaerobic hydrogen-producing micro-organisms were investigated using serum bottle assays. The enrichment pH values were controlled at 3, 4, 5, 7, 10, 11 and 12 with 1 N hydrochloric acid and 1 N sodium hydroxide. For each enrichment pH, the cultivation pH values were controlled at 5, 6 and 7. Based on the experimental results, hydrogen accumulation from sludge with acid or base enrichment is higher than that of the control. The hydrogen production potential of the sludge with acid or base enrichment is 200 and 333 times enhanced, compared with the control, when the enrichment pH is 10 and 3, respectively. The enhancement is due to a shortening of the micro-organisms' lag time which occurs at a proper cultivation-pH level. PMID- 11876413 TI - Molecular detection and diversity of polycyclic aromatic hydrocarbon-degrading bacteria isolated from geographically diverse sites. AB - Nineteen polycyclic aromatic hydrocarbon (PAH)-degrading bacteria were isolated from environmental samples in Kuwait, Indonesia, Thailand, and Japan by enrichment with either naphthalene or phenanthrene as a sole carbon source. Sequence analyses of the 16-S rRNA gene indicated that at least seven genera (Ralstonia, Sphingomonas, Burkholderia, Pseudomonas, Comamonas, Flavobacterium, and Bacillus) were present in this collection. Determination of the ability of the isolates to use PAH and its presumed catabolic intermediates suggests that the isolates showed multiple phenotypes in terms of utilization and degradation pathways. The large subunit of the terminal oxygenase gene (phnAc) from Burkholderia sp. strain RP007 hybridized to 32% (6/19) of the isolates, whilst gene probing using the large subunit of terminal oxygenase gene (pahAc) from Pseudomonas putida strain OUS82 revealed no pahAc-like genes amongst the isolates. Using three degenerated primer sets (pPAH-F/NR700, AJ025/26, and RieskeF/R), targeting a conserved region with the genes encoding the large subunit of terminal oxygenase successfully amplified material from 6 additional PAH-degrading isolates. Sequence analyses showed that the large subunit of terminal oxygenase in 4 isolates was highly homologous to the large subunit of naphthalene dioxygenase gene from Ralstonia sp. strain U2. However, we could not obtain any information on the oxygenase system involved in the naphthalene and/or phenathrene degradation by 7 other strains. These results suggest that PAH degrading bacteria are diverse, and that there are still many unidentified PAH degrading bacteria. PMID- 11876417 TI - Thermophilic biodesulfurization of naphthothiophene and 2-ethylnaphthothiophene by a dibenzothiophene-desulfurizing bacterium, Mycobacterium phlei WU-F1. AB - Naphtho[2,1-b]thiophene (NTH) is an asymmetric structural isomer of dibenzothiophene (DBT), and NTH derivatives can be detected in diesel oil following hydrodesulfurization treatment, in addition to DBT derivatives. Mycobacterium phlei WU-F1, which possesses high desulfurizing ability toward DBT and its derivatives over a wide temperature range (20-50 degrees C), could also grow at 50 degrees C in a medium with NTH or 2-ethylNTH, an alkylated derivative, as the sole source of sulfur. At 50 degrees C, the resting cells of WU-Fl degraded 67% and 83% of 0.81 mM NTH and 2-ethylNTH, respectively, within 8 h. By GC-MS analysis, 2-ethylNTH-desulfurized metabolites were identified as 2-ethylNTH sulfoxide, 1-(2'-hydroxynaphthyl)-1-butene and 1-naphthyl-2-hydroxy-1-butene, and it was concluded that WU-F1 desulfurized 2-ethylNTH through a sulfur-specific degradation pathway with the selective cleavage of carbon-sulfur bonds. Therefore, M. phlei WU-Fl can effectively desulfurize asymmetric organosulfur compounds, NTH and 2-ethylNTH, as well as symmetric DBT derivatives under high temperature conditions, and it may be a useful desulfurizing biocatalyst possessing a broad substrate specificity toward organosulfur compounds. PMID- 11876418 TI - Fungal contribution to in situ biodegradation of poly(3-hydroxybutyrate-co-3 hydroxyvalerate) film in soil. AB - The contribution of fungi to the microbial degradation of poly(3-hydroxybutyrate co-3-hydroxyvalerate) (PHBV) films in soil was studied. Various traces, cavities, and grooves observed on the dented surface of PHBV films demonstrated that the degradation was a concerted effect of a microbial consortium colonizing the film surface, including fungi, bacteria, and actinomycetes. The succession of microbial consortia in the soil around the PHBV films during the degradation showed a distinctive increase in the fungal population, resulting in its dominance. Comparison of the degradation ability of microbial strains isolated from soil where PHBV films were degraded, revealed that fungi showed the highest contribution to PHBV degradation, growing very rapidly along the film surface with their high degradation ability and then expanding their hyphae in a three dimensional manner. PMID- 11876419 TI - Assessment of metabolic properties and kinetic parameters of methanogenic sludge by on-line methane production rate measurements. AB - This report presents a new approach to studying the metabolic and kinetic properties of anaerobic sludge from single batch experiments. The two main features of the method are that the methane production is measured on-line with a relatively cheap system, and that the methane production data can be plotted as rate vs time curves. The case studies of specific methanogenic activity, biodegradability and toxicity tests here presented show that very accurate kinetic data can be obtained. The method is specifically useful in experiments in which strong changes in methane production occur, and it is proposed as a powerful tool to study methanogenic systems. Furthermore, the method is simple and could be implemented by industry in the routine analysis of sludge. PMID- 11876420 TI - Pure bacterial isolates that convert p-xylene to terephthalic acid. AB - Bacteria that grow on p-xylene, p-toluic acid, and terephthalic acid (TPA) were isolated from a wastewater bioreactor that is used to treat a waste stream that contains all three of these compounds. Although previously described aerobic bacteria degrade p-xylene by initially oxidizing a single methyl group to form p toluic acid and then cleaving the aromatic ring, some of the bacteria isolated during this study transformed p-xylene by oxidizing both methyl groups to produce TPA. PMID- 11876421 TI - Formaldehyde removal in synthetic and industrial wastewater by Rhodococcus erythropolis UPV-1. AB - Rhodococcus erythropolis strain UPV-1 is able to grow on phenol as the only carbon and energy source and to remove formaldehyde completely from both synthetic and industrial wastewater. The rate of formaldehyde removal is independent of either initial biomass or formaldehyde concentration. The presence of viable, intact cells is strictly necessary for this removal to take place. Discontinuous and continuous formaldehyde-feed systems were successfully tested with synthetic wastewater in shaken flasks. Once biodegradation was well established in model synthetic wastewater, a real wastewater sample was obtained from a local phenolic and melamine resin-manufacturing company. Incubation of biomass with this wastewater at subtoxic concentrations of formaldehyde resulted in the complete removal of the pollutant. Parameters, such as chemical oxygen demand and toxicity, were assessed as indicators of wastewater cleanup progress. PMID- 11876422 TI - An ATP-binding cassette multidrug-resistance transporter is necessary for tolerance of Gibberella pulicaris to phytoalexins and virulence on potato tubers. AB - The necrotrophic pathogen Gibberella pulicaris infects potato tubers through wounds that contain fungitoxic secondary metabolites such as the phytoalexins rishitin and lubimin. In order to colonize tuber tissue, the fungus must possess a mechanism to tolerate potato defense compounds. In this paper, we show that a gene, Gpabc1, that codes an ATP-binding cassette (ABC) transporter is required for tolerance to these phytoalexins and for virulence on potato. The Gpabc1 gene, isolated in the course of a differential cDNA screen, shares high sequence homology with the ABC1 gene of Magnaporthe grisea. G. pulicaris mutants deficient in Gpabc1 were still able to metabolize rishitin but lost their tolerance to this phytoalexin as well as their virulence on potato. These results strongly suggest that the Gpabc1-encoded ABC transporter is necessary for tolerance of G. pulicaris to rishitin and that this tolerance is required for virulence on potato. PMID- 11876423 TI - A signal transfer system through three compartments transduces the plant cell contact-dependent signal controlling Ralstonia solanacearum hrp genes. AB - Ralstonia solanacearum hrp genes encode a type III secretion system required for disease development in host plants and for hypersensitive response elicitation on non-hosts. hrp genes are expressed in the presence of plant cells through the HrpB regulator. This activation, which requires physical interaction between the bacteria and the plant cell, is sensed by the outer membrane receptor PrhA. PrhA transduces the plant cell contact-dependent signal through a complex regulatory cascade integrated by the PrhJ, HrpG, and HrpB regulators. In this study, we have identified two genes, named prhI and prhR, that belong to the hrp gene cluster and whose predicted products show homology with extracytoplasmic function sigma factors and transmembrane proteins, respectively. Strains carrying a mutation in prhIR show a delayed pathogenic phenotype toward host plants. PrhIR control the plant cell contact-dependent activation of hrp genes. prhIR gene expression is induced by a signal present in the plant cell coculture that is not PrhA dependent. Genetic evidence shows that PrhIR act upstream of PrhJ in the regulatory cascade, likely transducing the signal sensed by PrhA through the periplasm as described for signal transfer systems through three compartments. This is the first report of such a surface signaling mechanism activating pathogenicity determinants in response to a nondiffusible plant cell wall signal. PMID- 11876424 TI - CPR1: a gene encoding a putative signal peptidase that functions in pathogenicity of Colletotrichum graminicola to maize. AB - Colletotrichum graminicola causes anthracnose leaf blight and stalk rot of maize. We used restriction-enzyme mediated insertional (REMI) mutagenesis to identify a gene in this fungus that is required for pathogenicity to both stalks and leaves. The predicted polypeptide encoded by this gene, which we have named CPR1, is similar to a family of proteins that comprise one subunit of the eukaryotic microsomal signal peptidase. The nonpathogenic CPR1 REMI mutant contains a plasmid integration in the 3' untranslated region of the gene, 19 bp downstream from the stop codon. The result is a significant reduction in transcript levels in comparison to the wild type, perhaps as a result of increased transcript instability. We were unable to knock out the CPR1 gene, and it may be essential for viability. Microscopic examination of the REMI mutant on maize leaves revealed that it is fully capable of penetrating and colonizing host cells during the initial, biotrophic phases of the disease interaction but, unlike the wild type, it appears to be unable to switch to a necrotrophic mode of growth. We suggest that the CPR1 REMI mutant may be unable to secrete sufficient quantities of degradative enzymes to support that transition. The CPR1 REMI mutant provides us with a useful tool for future studies of the role of fungal protein transport in this important stalk rot disease of maize. PMID- 11876425 TI - Expression of genes encoding late nodulins characterized by a putative signal peptide and conserved cysteine residues is reduced in ineffective pea nodules. AB - Five nodulin genes, PsN1, PsN6, PsN314, PsN335, and PsN466, with reduced expression in ineffective nodules on the pea (Pisum sativum) mutant E135 (sym13) were characterized. They encode small polypeptides containing a putative signal peptide and conserved cysteine residues and show homology to the nodulins PsENOD3/14 and PsNOD6. For each gene, multiple bands were detected by genomic Southern analysis. Northern analysis showed that all five genes were expressed exclusively in nodules and that their temporal expression patterns were similar to that of the leghemoglobin (Lb) gene during nodule development. Their transcripts were localized predominantly from the interzone II-III to the distal part of nitrogen-fixing zone III in effective nodules, resembling the Lb gene. However, transcripts in ineffective E135 nodules were localized in narrower regions than those in the effective nodules. These results indicate that these nodulins are abundant in pea nodules and that their successive expression during nodule development is associated with nitrogen-fixing activity. PMID- 11876426 TI - Attachment to roots and virulence of a chvB mutant of Agrobacterium tumefaciens are temperature sensitive. AB - Agrobacterium tumefaciens chvB mutants are unable to produce beta-1,2 glucan. They are nonattaching and avirulent and show reduced motility at room temperature. At lower temperatures (16 degrees C), chvB mutants became virulent on Bryophyllum daigremontiana and Lycopersicon esculentum and were able to attach to L. esculentum, Arabidopsis thaliana, Daucus carota, and Tagetes erecta roots. The mutant bacteria also recovered wild-type motility at lower temperatures. Two other nonattaching mutants of A. tumefaciens, AttR and AtrA, were unaffected by the lowered temperature, remaining nonattaching and avirulent. PMID- 11876427 TI - Patterns of pectin methylesterase transcripts in developing stem nodules of Sesbania rostrata. AB - Differential display was applied to the early stages of the interaction between the tropical legume Sesbania rostrata and its microsymbiont Azorhizobium caulinodans ORS571. An upregulated clone that is similar to pectin methylesterase encoding genes was isolated (Srpmel). The full-length sequence of Srpme1 was used to localize PME transcripts in situ during S. rostrata stem-nodule development. Several expression patterns were distinguished, hinting at general roles in vascular tissue development and cell division or expansion and at symbiosis specific functions, such as uninfected cell differentiation. PMID- 11876428 TI - A new pathotype of Pea seedborne mosaic virus explained by properties of the P3 6k1- and viral genome-linked protein (VPg)-coding regions. AB - A fourth pathotype of Pea seedborne mosaic virus, a member of the genus Potyvirus, was identified by analysis of the infection profile on a panel of Pisum sativum lines. The new pathotype, designated P-3, was able to overcome resistance specified by the sbm-1 resistance gene but could not overcome resistance specified by the sbm-2 resistance gene. This infection profile distinguished P-3 from previously described pathotypes, P-1, P-2, and P-4. Analysis of chimeric viruses demonstrated that properties of the P3-6k1- and viral genome-linked protein (VPg)-coding regions accounted for the infection profile of the new pathotype. PMID- 11876430 TI - There is perhaps no area of science that holds more promise or elicits more fear than genetic research. PMID- 11876429 TI - Fdb1 and Fdb2, Fusarium verticillioides loci necessary for detoxification of preformed antimicrobials from corn. AB - Fusarium verticillioides is a fungus of significant economic importance because of its deleterious effects on plant and animal health and on the quality of their products. Corn (Zea mays) is the primary host for F. verticillioides, and we have investigated the impact of the plant's antimicrobial compounds (DIMBOA, DIBOA, MBOA, and BOA) on fungal virulence and systemic colonization. F. verticillioides is able to metabolize these antimicrobials, and genetic analyses indicated two loci, Fdb1 and Fdb2, were involved in detoxification. Mutation at either locus caused sensitivity and no detoxification. In vitro physiological complementation assays resulted in detoxification of BOA and suggested that an unknown intermediate compound was produced. Production of the intermediate compound involved Fdbl, and a lesion in fdb2 preventing complete metabolism of BOA resulted in transformation of the intermediate into an unidentified metabolite. Based on genetic and physiological data, a branched detoxification pathway is proposed. Use of genetically characterized detoxifying and nondetoxifying strains indicated that detoxification of the corn antimicrobials was not a major virulence factor, since detoxification was not necessary for development of severe seedling blight or for infection and endophytic colonization of seedlings. Production of the antimicrobials does not appear to be a highly effective resistance mechanism against F. verticillioides. PMID- 11876431 TI - Perceptions of alternative therapies available for women facing hysterectomy or menopause. AB - Women's perceptions of alternative therapies available for managing symptoms related to menopause and to the need for hysterectomy were explored. Data are reported from 17 focus groups of peri- and menopausal aged women (n = 82) living in two southern U.S. coastal counties. Analysis showed that emergent themes did not vary by race or surgery experience. Herbal preparations, vitamin supplements, "healthy living" (diet & exercise), and mind/body practices (prayer & "mental healing") were mentioned as possible alternatives for managing symptoms. Participants more frequently identified "other" conventional medical approaches- prescription medication, laser surgery, dilation and curettage, and "watchful waiting"--as alternatives to hysterectomy. Alternative therapies are linked to women's desire to gain control over their own health care decisions and practices. PMID- 11876432 TI - I want to know: exploring how older women acquire health knowledge after a myocardial infarction. AB - The purpose of this descriptive naturalistic study was to: (a) explore how older women not participating in outpatient cardiac rehabilitation (CR) acquire health knowledge regarding secondary prevention measures after a myocardial infarction (MI), and (b) describe women's perceived barriers to acquiring this knowledge. Women (N = 15), ages 66-88, participated in semi-structured in-depth interviews. Data analysis resulted in four global categories: (a) previous learning, (b) self knowledge, (c) actively seeking knowledge from others, and (d) information provided by others. Barriers to acquiring health knowledge after MI without the support of outpatient CR were also identified. PMID- 11876433 TI - A prediction model for polypharmacy: are older, educated women more susceptible to an adverse drug event? AB - INTRODUCTION: This study attempts to account for variation in the number of prescription medications concurrently consumed in geriatric populations. METHODS: Data were selected from The National Health and Nutrition Examination Survey, III, 1988-1994 (NHANES III), Adult Household Questionnaire, a complex, multistage, clustered sampling of civilian, non-institutionalized populations, which included a volunteer sample of 5,249 individuals aged 65 and older who participated in NHANES III survey, representing four broad geographic regions and twelve states. RESULTS: Age, income, and educational level accounted for nine percent of the variation in the number of prescription medications concurrently taken (p < 0.0001); though non-significant, there were gender differences regarding polypharmacy; in addition, there were significant differences with respect to educational levels by region and age by region (p < 0.05). CONCLUSIONS: Educational level accounted for the most variation in polypharmacy. Women, on average, consumed more prescription medications than men. Older, more educated women may be most likely to engage in polypharmaceutical consumption, suggesting a greater likelihood of an adverse drug event. PMID- 11876434 TI - Women's retirement, work and life paths: changes, disruptions and discontinuities. AB - In this qualitative study we examine women's perceptions and interpretations of the impact of retirement on their life experience, whether retirement was encountered from unexpected workplace redundancies, through their own decisions, or those of their partner. In the first instance we examine traditional definitions and meanings of retirement and their relevance to women's experience. Any theoretical explanation that work is central to defining core identity and social integration is challenged. Secondly, this study provides some alternative ways to examine changes in work, disruptions, and discontinuities in the context of women's life paths. Whereas specific identities and roles may be important, being able to contribute to social life, whether through paid or unpaid work or through self-interests, may be more appropriate determinants of societal integration, than identities that are formed and maintained by a production oriented society. PMID- 11876435 TI - Support groups for older victims of domestic violence. AB - A 1997 nationwide (US and Canada) search to identify support groups for older victims of domestic violence located 16 sponsored by domestic violence programs and 14 sponsored by aging services. Interviews with group leaders indicated more similarities than differences between the two types of sponsorship in group purpose, leadership, numbers served, content of support group sessions, and success in accomplishing goals. Resistance of elders to participate in a group experience was cited by leaders as a major barrier. Recommendations for future groups include insuring accessibility of meeting site; using a leader and co leader, at least one of whom is older or familiar with aging issues; allocating resources for recruitment; and seeking a steady source of funding. A policy of collaboration among the state's domestic violence coalition, state unit on aging, adult protective services, and victim assistance program may help in promoting support group development and utilization. PMID- 11876436 TI - Determination of amezinium in plasma by RP HPLC and its application to bioequivalence studies. AB - A modified method is described for the determination of 4-amino-6-methoxy-l phenylpyridazinium methyl sulfate (amezinium metisulfate) in plasma and in the dosage form. The method involves a sample preparation using SPE and liquid-liquid extraction and subsequent determination by high performance liquid chromatography. Retention time for amezinium was 3.2 min and there was no interference from plasma or amezinium metabolites. The method was applied for the drug determination during comparative studies of two different tablet formulation of amezinium. PMID- 11876437 TI - Estimation of buspirone-bovine serum albumin binding by affinity capillary electrophoresis. AB - Drug-protein binding is an important process in pharmacokinetic phase of drug action. Capillary electrophoresis was employed, specifically the Hummel-Dreyer method and Scatchard analysis, to study the interactions of an anxiolytic drug, buspirone, with pure bovine serum albumin (BSA) and with BSA present in the human recombinant 5-HT(1A) serotonin receptor preparation. The binding constant of buspirone with BSA determined in free BSA solution was K = 5.55 x 10(4) M(-1) whereas its value with BSA present in the serotonin receptor preparation was K = 5.57 x 10(4) M(-1). The method was found to be inadequate for measuring the specific binding interactions between buspirone and the 5-HT(1A) receptor in the preparation employed. PMID- 11876438 TI - Determination of 4-aminophenol impurities in selected pharmaceutical preparations by HPLC method with amperometric detection. AB - 4-Aminophenol (4-APh) is the main impurity present in preparations containing paracetamol. Using the cyclic voltammetry and differential pulse voltammetry methods, the electrode behaviour of 4-APh has been studied in various non buffered and buffered solutions at glassy carbon and golden electrodes. By means of the high-performance liquid chromatography with amperometric detection (HPLC EC), the dependency of current intensities of the 4-APh peaks on the potential in the range (0-600 mV in the pH range 2-5, the ionic strength of mobile phase ranging from 0.01 mol (-1) to 0.20 mol l(-1) LiCl, has been studied. While employing the HPLC-EC method in this work, a glassy carbon electrode was used as the amperometric detector. The optimal potential selected in this research was +325 mV. It was found that a 0.05 mol l(-1) LiCl solution, containing 18% of methanol, at pH 4.0 adjusted with orthophosphoric acid, is suitable for the separation of 4-APh and paracetamol from each other and from other pharmaceutical excipients present in tablets. By using the elaborated HPLC-EC method, the content of 4-APh (at concentrations from 4 ng ml(-1)) in the paracetamol from Aldrich and in tablets from different producers, containing 500 mg of paracetamol in a tablet was found. Statistical evaluation of the obtained results has shown that the proposed HPLC-EC method for the determination of 4-APh is characterized by a good accuracy and precision (RSD about 6%) and can be applied to routine investigations of pharmaceutical preparations in the form of tablets. PMID- 11876439 TI - Identification and determination of selected medicines reducing hypertension by densitometric and gas chromatographic methods. AB - Conditions have been elaborated for the identification and quantitative determination of captopril and enalapril in pharmaceutical preparations by the densitometric method in the concentration range 5-50 microg, and of enalapril by gas chromatography in the concentration range 0.5-3.5 mg/ml. The separation of the preparations was performed using methanol. In the densitometric method the determination of captopril was done at lambda = 212 nm, while for enalapril at lambda = 210 nm. In the gas chromatographic method a DB-17 capillary column was used as well as a flame ionization detector. Perindopril was used as the internal standard. PMID- 11876440 TI - Determination of hydrochlorothiazide, triamterene and propranolol hydrochloride by the spectrophotometric method and high-performance liquid chromatography (HPLC). AB - Spectrophotometric and chromatographic (HPLC) methods for determination of hydrochlorothiazide, triamterene and propranolol hydrochloride were elaborated. Both methods were appropriate for the determination of three compounds in pharmaceutical preparations containing their mixtures. Both the elaborated methods for the determination of the studied compounds give comparable results and can successfully be applied to the assay in their mixtures occurring in the composition of pharmaceutical preparations. PMID- 11876441 TI - Bioavailability of tramadol hydrochloride from tramadol--capsules 50 mg. AB - The aim of this study was to investigate the bioavailability of tramadol hydrochloride after oral administration of Tramadol--50 mg capsules, made in Synteza Pharmaceutical-Chemical Company in Poznan. As a reference preparation of Tramadol was used Tramal--50 mg capsules, (Grunenthal, Germany). The preparations were investigated in 20 healthy volunteers, according to a randomised two-way, cross-over design in the fasted state. Blood samples for determination of tramadol plasma concentrations were collected at pre-defined time points up to 24 h following drug administration. A washout period of one week separated both treatment periods. Tramadol plasma concentrations were determined by means of a validated HPLC method (fluorescence detector, verapamil as an internal standard). Values of 1,226.4 ng h/ml (Tramadol) and 1,397.01 ng x h/ml (Tramal) for the parameter AUC(0-infinity) demonstrate a nearly identical extent of drug absorption. Maximum concentrations--Cmax (217.81 ng/ml and 246.0 ng/ml) and time to reach maximum plasma concentration--Tmax (2.14 and 2.31 h) achieved for Tramadol and reference preparation did not differ significantly. CONCLUSIONS: the bioavailability of tramadol hydrochloride after administration of Tramadol is the same as after administration of Tramal, whose clinical efficacy was tested before. PMID- 11876442 TI - Synthesis and evaluation of 2-chloroethylnitrosoureas of substituted naphthalimides as mixed-function anticancer compounds. AB - New mixed function anticancer compounds as 2-chloroethylnitrosoureas of substituted naphthalimides represented by bromonap-NU 4a and chloronap-NU 4b, have been synthesized from 4-bromo- and 4-chloro-l,8-naphthalic anhydride, respectively following a 3-step process. Their chemical alkylating activity compared with nor -HN2 indicated that they possess greater alkylating activity than the latter. Their antitumour efficacies were assessed in vivo in two murine ascites tumours, namely Ehrlich ascites carcinoma (EAC) and Sarcoma-180 (S-180) by measuring the increase in median survival times (MST) of drug treated (T) over untreated control (C) mice. Two standard clinical drugs namely endoxan (cyclophosphamide) and 5-fluorouracil (5-FU) were used as positive controls for comparison. Both of them have displayed substantial and reproducible antitumoral activity in these tumours comparable with 5-FU. These were further screened in vitro in 6 different human tumour cell lines but no significant activity was observed in those lines. PMID- 11876443 TI - Synthesis of 1,4-disubstituted 2-methylpiperazine derivatives, new 5-HT(1A) receptor ligands. AB - Preparation of some new 1,4-substituted 2-methylpiperazine derivatives is reported. The influence of structural modifications on their affinity to 5-HT(1A) receptors is discussed. Compounds were synthesised by the reaction of 2 methylpiperazine with 2-chloropyrimidine or 2-chloroquinoline followed by condensation with 1,4 dibromobutane. The resulting quaternary ammonium salts after the reaction with an imide gave the respective final products. PMID- 11876444 TI - Flavonoids from the flowers of Prunus spinosa L. AB - Eight flavonoids were isolated from the flowers of Prunus spinosa: kaempferol, quercetin, kaempferol 3-O-alpha-L-arabinofuranoside, quercetin 3-O-alpha-L arabinofuranoside, kaempferol 3-O-alpha-L-ramnopyranoside, kaempferol 7-O-alpha-L ramnopyranoside, kaempferol 3-O-beta-D-xylopyranoside, kaempferol 3-O-(2''-E-p coumaroyl)-alpha-L-arabinofuranoside. The last three have been found for the first time in this plant. The structure of the compounds was determined by means of chemical and spectral methods (UV, IR, LSI MS, 1H NMR, 13C NMR). PMID- 11876445 TI - Comparative analysis of phenolic acids in mistletoe plants from various hosts. AB - Phenolic acids present in mistletoe plants collected from various hosts were analysed with the use of HPLC. The following numbers of compounds were found in the mistletoe plant material gathered from respective hosts: Sorbus aucuparia- 12 compounds; Acer plantanoides--14 compounds: Malus domestica, Pyrus communis and Populus nigra--13 compounds each; Quercus robur--15 compounds. Altogether 21 phenolic acids were chromatographically identified in the tested material. The compounds were either free or combined as esters or glycosides. Comparative chromatography revealed qualitative differences in the investigated compounds between the various plant materials. For example o-coumaric acid was only found in mistletoe hosted by Quercus robur. Digallic acid was only found in the plant material hosted by Acer plantanoides. Qualitative and quantitative composition of mistletoes hosted by Malus domestica and Pyrus communis showed considerable similarities as far as phenolic acids were concerned. Moreover. vanillic acid. absent in all other batches of plant material, seemed to be characteristic of the above mistletoes. Quantitative HPLC analysis demonstrated a considerable content of salicylic acid (39.55 mg%) in mistletoe hosted by Sorbus aucuparia. Apart from the above material, this compound was only present in small quantities in plants hosted by Populus nigra (15.63 mg%) and Quercus robur (2.63 mg%). PMID- 11876446 TI - Synthetic and bioorganic investigations of 2-cyclohexylthiocarbonyl( 1,2,3,6,7,11B)hexahydro-4H-pyrazino [2-1a] isoquinoline-4-thione, a new dithione mimic of the universal anthelminthic Praziquantel. AB - As a continuation for our previous approaches to establish structure antischistosomal activity relationship (SAR) among some new rationally synthesized analogues of praziquantel, herein a new C-4 and C-12 dithione mimic of the drug namely, 2-cyclohexylthiocarbonyl (1, 2, 3, 6, 7, 11b) hexahydro-4H pyrazino[2-la]isoquinoline-4-thione (II) was synthesized and antischistosomally investigated (mice infected with S. masoni cercariae). Further, some significant biochemical and toxicological parameters for both the control and the dithione II treated mice, particularly the total serum and liver proteins, liver enzymes, serum total lipids, cholesterol, triglycerides, albumin, globulins and creatinine, were assayed. The determined induced amino acid profile of liver protein hydrolysate could indicate a close similarity of the working biological mechanism for both I and II. Comparable to praziquantel, the dithione II was found, still promisingly antischistosomally active (approximately 70% of I, collective average activity, based on 500 mg II/kg mouse body weight). Equally, generally tolerant toxicity parameters for liver and kidney functions could be attributed. Due to the still absence of quasi-potent praziquantel candidates since its discovery (1975), the dithione II could be considered as an interesting anthelminthic candidate susceptible for further profound studies and structure modulations. In this context, some perspectives were also suggested. PMID- 11876447 TI - Evaluation of alpha-tocopherol, probucol and ascorbic acid as suppressors of digoxin induced lipid peroxidation. AB - Protective effects of three free radical scavengers, tocopherol (TOC), probucol (PR) and ascorbic acid (AA), on cardiotonic glycoside digoxin (DIG) induced lipid peroxidation in goat liver homogenate, have been studied by measuring malondialdehyde and glutathione contents as indicator parameters. The level of reduced glutathione decreased vis-a-vis malondialdehyde content increased in the drug treated samples in comparison with the controls. This suggests that DIG may have significant lipid peroxidation induction capacity. Considering lipid peroxidation as a toxicity mediating process, this may be related to the toxic potential of the drug. When the liver homogenate samples were incubated with antioxidant (TOC/PR/AA) in conjunction with the drug (DIG), lipid peroxidation was suppressed as indicated by increased level of reduced glutathione and decreased level of malondialdehyde in comparison with those of drug treated samples. This indicates that TOC, PR and AA may have considerable suppressive action on DIG induced lipid peroxidation. Thus, these antioxidants merit further extensive study to explore their potential in reducing DIG induced toxicity that may be mediated by free radical mediated process. PMID- 11876448 TI - Qualitative and quantitative chromatographic investigation of flavonoids in Bellis perennis L. AB - The qualitative and quantitative analysis of flavonoids in the flowers and leaves of naturally growing B. perennis and in the flowers of cultivated variations 'rosea' and 'rubra' were carried out. The qualitative analysis was carried out by TLC and compared with flavonoid compounds isolated from flowers of common daisy. The quantitative determinations were carried out by Christ-Muller's method and by HPLC after acid hydrolysis. Similar flavonoids were found in all the flowers, while differences were noted in the flavonoid composition of the leaves. The flavonoid contents were higher in the flowers than in the leaves. PMID- 11876449 TI - Synthesis and in vitro tuberculostatic activity of Co (II), Cu (II) and Ni (II) complexes of dialdehyde starch dithiosemicarbazone. AB - Co (II), Cu (II) and Ni (II) complexes of starch dialdehyde dithiosemicarbazone (DASTSC) of low (approximately 15%) degree of oxidation were prepared and their tuberculostatic activity was tested in vitro against isoniazid-sensitive and isoniazid-resistant strains of Mycobacterium tuberculosis. In 10-week lasting tests the DASTSC complex with Ni (II) was tuberculostatic to a similar extent as was the free ligand, whereas the Co (II) and Cu (II) complexes inhibited the growth of M. tuberculosis more efficiently. It was also proved that the corresponding non-co-ordinated metal salts were inactive to both strains of M. tuberculosis. PMID- 11876450 TI - Pertussis: a continuing hazard for healthcare facilities. PMID- 11876451 TI - Nasal carriage of Staphylococcus aureus among patients receiving allergen injection immunotherapy: associated factors and quantitative nasal cultures. AB - OBJECTIVE: To compare the prevalence of nasal Staphylococcus aureus carriage among outpatients receiving allergen-injection immunotherapy with the prevalence among healthy controls and to determine predictors of nasal S. aureus carriage. DESIGN: Survey. SETTING: Allergy clinic of a university hospital. PARTICIPANTS: A volunteer sample consisting of 45 outpatients undergoing desensitization therapy and 84 first- and second-year medical students. RESULTS: The nasal S. aureus carriage rate was significantly higher among patients (46.7%) than among students (26.2%; P=.019). In a multivariate model adjusted for age and gender, the presence of atopic dermatitis or eczema was the only independent predictor of nasal S. aureus carriage (odds ratio [OR], 4.4; 95% confidence interval [CI95], 1.2-16.0; P=.02). The only other participant characteristic associated with nasal S. aureus carriage was immunotherapy with allergen injections (OR, 1.98; CI95, 0.7-6.0), but this association did not reach statistical significance (P=.23). The probability of nasal S. aureus carriage was 88.9% for patients receiving allergen injections and having atopic dermatitis or eczema, and 36.1% for patients receiving allergen injections without atopic dermatitis or eczema. CONCLUSIONS: Patients undergoing desensitization have a higher nasal carriage rate of S. aureus. However, factors other than the regular use of needles, and in particular abnormalities related to the atopic constitution of these patients, may predispose this population for S. aureus carriage. PMID- 11876452 TI - Nosocomial infections in geriatric long-term-care and rehabilitation facilities: exploration in the development of a risk index for epidemiological surveillance. AB - OBJECTIVE: To compute a risk index for nosocomial infection (NI) surveillance in geriatric long-term-care facilities (LTCFs) and rehabilitation facilities. DESIGN: Analysis of data collected during the French national prevalence survey on NIs conducted in 1996. Risk indices were constructed based on the patient case mix defined according to risk factors for NIs identified in the elderly. SETTING: 248 geriatric units in 77 hospitals located in northern France. PARTICIPANTS: All hospital inpatients on the day of the survey were included. RESULTS: Data from 11,254 patients were recorded. The overall rate of infected patients was 9.9%. Urinary tract, respiratory tract, and skin were the most common infection sites in both rehabilitation facilities and LTCFs. Eleven risk indices, categorizing patients in 3 to 7 levels of increasing NI risk, ranging from 2.7% to 36.2%, were obtained. Indices offered risk adjustment according to NI rate stratification and clinical relevance of risk factors such as indwelling devices, open bedsores, swallowing disorders, sphincter incontinence, lack of mobility, immunodeficiency, or rehabilitation activity. CONCLUSION: The optimal index should be tailored to the strategy selected for NI surveillance in geriatric facilities in view of available financial and human resources. PMID- 11876453 TI - Hepatitis c virus infection in employees of a large university hospital in Israel. AB - OBJECTIVE: To assess whether hospital work constitutes a risk factor for hepatitis C virus (HCV) infection among employees of a large hospital in Israel. DESIGN: Seroprevalence survey. SETTING: A 1,006-bed, tertiary-care university hospital in Jerusalem. PARTICIPANTS: All 5,444 employees (18-65 years old) were eligible; 4,287 (79%) participated in the survey. METHODS: Sera were tested for antibodies to HCV (anti-HCV) using a third-generation enzyme immunoassay. A third generation strip immunoblot assay was used for confirmation. Participants were interviewed regarding their occupational history, and they completed a self administered questionnaire covering history of non-occupational exposure to blood and country of birth. Other demographic information was obtained from the personnel department. Rates and odds ratios (ORs) were calculated, and multivariate logistic-regression analyses were performed to adjust for potential confounding variables. RESULTS: Anti-HCV was found in 0.9% of employees (37/4,287; 95% confidence interval, 0.6-1.1), ranging from 0.1% among those born in Israel to 5.7% among those born in Central Asia. After age, gender, social status, country of birth, and history of blood transfusion were controlled for in a logistic regression, occupational exposure to blood > or = 10 years was significantly associated with the presence of antibodies (OR, 2.6; P=.01). Presence of anti-HCV also was associated with country of birth (range: Israel OR, 1; West OR, 3.8 [P=.1]; Central Asia OR, 48.6 [P<.0001]) and history of blood transfusion (OR, 2.7; P=.01). No significant associations were found between anti HCV and age, gender, social status, history of tattoo, acupuncture, current occupation, department, exposure to blood in current occupation, adherence to safety precautions, or history of percutaneous injury. The association with length of exposure was stronger (OR, 3.6; P=.01) when the same logistic regression was run excluding the outlier ethnic group of Central Asia. CONCLUSIONS: Hospital work does not seem to constitute a major risk factor for HCV infection in Israel today. A higher prevalence of anti-HCV among employees with longer versus shorter lengths of occupational exposure may be due to a cumulative effect of exposure over the years. Infection control efforts in recent years may have contributed to this association. PMID- 11876454 TI - Nosocomial infections in a children's hospital in Argentina: impact of a unique infection control intervention program. AB - OBJECTIVE: To assess the efficacy of parental education and use of parents as nursing assistants on reducing nosocomial infections. DESIGN: Prospective study. METHODS: Active surveillance for nosocomial infections was performed on two wards. On ward A, parents were educated about infection control practices and assisted nursing staff with routine tasks, so that nursing personnel could focus their efforts on procedures with higher risk of infection. Parental assistance was not sought on ward B, the comparison ward. RESULTS: From October 1990 through September 1991, 1,081 patients were admitted to wards A (470) or B (611). The over-all nosocomial infection rate was 7.1 per 100 admissions; the nosocomial infection rate was significantly higher on ward B than ward A (63/611 vs 14/470; P<.001). Multivariate analysis identified risk factors for nosocomial infection on the two wards as age <2 years (P=.01), malnutrition (P=.005), duration of hospitalization (P<.001), ward B hospitalization (P=.003), and ward cleanliness score (P=.009); the distribution of patients with these factors was similar on the two wards. CONCLUSIONS: Our data suggest that parental infection control education and recruitment to relieve nursing staff of routine low-risk procedures are economical and easily implemented measures to reduce nosocomial infections in hospitals with limited personnel resources in the developing world. PMID- 11876455 TI - Neonatal bacteremia: patterns of antibiotic resistance. AB - OBJECTIVE: To determine the incidence and evaluate the antimicrobial susceptibility patterns of bacterial infections in our neonatal units. DESIGN: Retrospective surveillance study. SETTING: The neonatal units of the Hadassah University Hospitals, Jerusalem, Israel. PATIENTS: All newborns admitted from January 1994 through February 1999. METHODS: The records of all patients with positive blood and cerebrospinal fluid cultures were reviewed. Bacteremia was considered early-onset (vertical) when occurring within the first 72 hours of life and late-onset (nosocomial) when occurring later. The prevalence and antibiotic-resistance patterns of vertically transmitted and nosocomially acquired strains were compared and studied over time. RESULTS: 219 of 35,691 newborn infants had at least one episode of bacteremia (6.13/1,000 live births). There were 305 identified organisms, of which 21% (1.29/1,000 live births) were considered vertically transmitted and 79% nosocomially acquired. The most common organism causing early-onset disease (29.2%) was group B streptococcus (0.38/1,000 live births), whereas coagulase-negative staphylococci (51%) were the most prevalent in late-onset disease. All gram-positive bacteria were susceptible to vancomycin. Most gram-positive organisms other than staphylococci were susceptible to ampicillin. Gram-negative organisms represented 31% of all isolates. Generally, there was a trend of increasing resistance to commonly used antibiotics among nosocomially acquired gram-negative organisms, compared to those vertically transmitted, with statistically significant differences for ampicillin and mezlocillin (P<.05 and P<.01, respectively). Over the years, a trend toward an increasing resistance to antibiotics was observed among gram negative organisms. CONCLUSIONS: The trend of increasing bacterial resistance to commonly used antibiotics necessitates the implementation of a rational empirical treatment strategy, based on local susceptibility data, reserving certain agents for emerging resistant pathogens. PMID- 11876456 TI - Nosocomial infections in pediatric cardiac surgery, Italy. AB - OBJECTIVE: To evaluate the incidence of nosocomial infection (NI) in pediatric patients who received cardiothoracic surgery and to identify possible associated risk factors. DESIGN: Prospective observational study. SETTING: The cardiac surgery and cardiac intensive care units at the Regina Margherita Children's Hospital, Turin, Italy. PATIENTS: All patients who underwent surgery from July 20, 1998, to July 19, 1999, were enrolled, except patients with operative catheterization only. METHODS: Clinical data were collected daily from July 20, 1998, to July 19, 1999. NIs were diagnosed according to US Centers for Disease Control and Prevention criteria. RESULTS: 104 patients were included in the present study, 80 (76.9%) of whom underwent extracorporeal circulation. The NI ratio was 48.1% (50/104); the percentage of patients with NI was 30.8% (32/104): 23.1% developed one infection, 7.7% two or more. The rate of NI was 2.17 per 100 days of hospitalization (50/2,304). The most common pathogen was Pseudomonas aeruginosa. Important risk factors were length of preoperative admission >5 days, total length of admission >10 days, open chest during postoperative phase, and cyanotic heart disease. There was a significant association between sepsis and central venous catheterization for 3 days or more. Rate of sepsis was 19 per 1,000 catheter days (16/852). CONCLUSION: NIs represent a frequent complication for children who undergo heart surgery. Based on our data, we suggest decreasing the preoperative stay as much as possible. The higher NI incidence in patients with an open chest postoperatively suggests that an alternative antibiotic strategy should be considered for these patients. PMID- 11876457 TI - Epidemiology of biological-exposure incidents among Spanish healthcare workers. AB - OBJECTIVE: To determine the frequency and the epidemiological characteristics of biological-exposure incidents occurring among healthcare personnel. DESIGN: Prospective surveillance study. SETTING: Participating Spanish primary-care and specialty centers from January 1994 to December 1997. PARTICIPANTS: 70 centers in 1994, 87 in 1995, 97 in 1996, and 104 in 1997. METHODS: Absolute and relative frequencies were calculated for several variables (position held, area of care, type of injuring object, activity, etc) and for the different categories of each variable. RESULTS: There were 20,235 registered incidents. Annual incidence rates were as follows: 1994, 51 per 1,000; 1995, 58 per 1,000, 1996, 54 per 1,000; and 1997, 59 per 1,000. Mean age of accident victims was as follows: 1994, 35.68 (standard deviation [SD], 16.26); 1995, 33.6 (SD, 11.9); 1996,38.2 (SD, 17.27); and 1997, 36.7 (SD, 16.33) years. Of the 20,235 incidents, 15,860 (80.7%) occurred to women; 50% (9,833) accidents were among nursing staff. The type of incident most frequently reported was percutaneous injury (81.1%). The highest frequency of accidents was seen in medical and surgical areas (28% and 25.6%, respectively). Blood and blood products were the most commonly involved material (87.6%). Administration of intramuscular or intravenous medication was the activity associated with the highest accident rate (20.3%). The most frequent immediate action in response was rinsing and disinfecting (65.6%). CONCLUSIONS: The incident registry was highly stable in terms of incidence rates over the observation period and served to highlight the large number of incidents recorded each year. The potential implications of the results are the need to explore reasons for increased exposures in certain areas, with the aim of focusing prevention efforts, and, similarly, to establish the factors associated with diminished incidence rates to model successful measures. PMID- 11876458 TI - Azithromycin prophylaxis during a hospitalwide outbreak of a pertussis-like illness. AB - A questionnaire regarding tolerability and adherence was administered for 5 days to hospital employees who received azithromycin prophylaxis during a hospitalwide outbreak of a pertussis-like illness. Analysis of the 239 responses from those having received prophylactic azithromycin determined that it was well tolerated and accounted for a minimal loss of days worked; 81.5% were fully adherent with the regimen. PMID- 11876460 TI - FDA approves first nucleic acid test for HIV and HCV. PMID- 11876459 TI - Nasal MRSA colonization of AIDS Patients cared for in a Brazilian university hospital. AB - Weekly culture surveillance was conducted over a 2-year period to determine the incidence of methicillin-resistant Staphylococcus aureus nasal colonization among acquired immunodeficiency syndrome patients cared for in a day-care unit and in an infectious diseases unit. Analysis of genomic DNA profiles showed a predominant pattern in both units. PMID- 11876461 TI - Circadian and thermoregulatory responses hold clinical clues. PMID- 11876462 TI - Circadian rhythmicity and homeostatic stability in thermoregulation. AB - Stability and circadian variation in core body temperature (Tc) were believed to be homeostatic responses until well into the 20th century. Defense of a narrow thermoneutral range was well documented, whereas circadian oscillations were attributed to episodic biochemical and environmental stimuli or chronological stressors in life routines. Research in thermal physiology has illuminated several of the "black boxes" in the understanding of temperature regulation, and advances in chronobiology have shattered old paradigms. While these discoveries are still evolving, existing information provides valuable clues about physiological responses to heat loss or over-heating that could improve clinical assessment and intervention. Discoveries that circadian rhythm of Tc is regulated by an endogenous "clock" and is remarkably stable have helped to make it the most widely used circadian indicator. More recently, Tc was found to exert its own cyclic rhythm under free-running conditions. While some investigators claim that circadian and homeostatic processes are independent, there are conditions in which clinical distinctions are less clear. This overview reviews contemporary scientific findings about circadian and homeostatic processes in thermoregulation. Examples are drawn from human and animal research. Physiological responses and mechanisms are explained in relation to their relevance to clinical treatment or health care. Gaps in existing research and application are discussed. PMID- 11876463 TI - Measuring circadian temperature rhythm. AB - Experimental control and mathematical techniques increase confidence that results of circadian temperature rhythm studies reflect true changes in the circadian timing system versus coupling with exogenous synchronizers. Masking effects represent confounding influences in studies that are concerned with the endogenous temperature rhythm. Because it is technically difficult to measure directly the behavior of the endogenous timing system, marker rhythms are used as proxy measures. However in addition to entraining, the external environment exerts a direct masking effect on the monitored rhythm. Methods for measuring circadian temperature rhythm are reviewed in this article. Constant routine, forced desynchrony, and purification methods represent attempts, at an experimental or mathematical level, to remove masking effects and more accurately capture the endogenous circadian temperature rhythm. Exogenous factors have not been subjected to the same scrutiny as the endogenous features of circadian temperature rhythm. But it is the environmental context, the extent to which the endogenous features are adaptively modified by the field environment, that will ultimately determine the biological value of circadian temperature rhythm to the organism. Thus, nurse investigators are encouraged to use rigorous methods to study both endogenous circadian temperature rhythm and exogenous rhythms. PMID- 11876464 TI - Effect of genetic obesity on thermoregulatory activity responses to inversion of the light/dark cycle. AB - In this pilot work, the authors compared the circadian rhythm responses of 3 month-old female fatty Zucker rats with those of lean Zucker rats and Sprague Dawley rats to reversal of the light/dark (LD) cycle. Core temperature and spontaneous cage activity were continuously monitored by implanted microtelemetry devices prior to and for 5 days following complete LD cycle reversal. By the 5th day after LD reversal, temperature rhythm nadir had phase-advanced 10.5 hours in the lean Zucker rats, 10.8 hours in the Sprague-Dawley rats, and only 3.8 hours in the fatty Zucker rats. Similarly, total activity increased in lean Zucker rats and Sprague-Dawley rats after LD reversal but declined in the fatty Zucker rat during the same time. Results of this study show that obese Zucker rats displayed an impaired ability to reentrain circadian rhythms for temperature and activity when compared to lean Zucker rats and Sprague-Dawley rats. These findings suggest that dysfunction in the circadian pacemaker previously shown to manifest itself by 43 weeks of age in fatty Zucker rats may already be present at 3 months of age and suggest that altered thermoregulation may play a role in the development of obesity in this animal model of genetic obesity. PMID- 11876465 TI - Reducing disruption of circadian temperature rhythm following surgery. AB - Temperature and other circadian rhythms are disrupted following surgery and other traumatic events. During recovery, coordination between temperature rhythms and other rhythmic physiologic processes is reduced. Studies of animals and humans have shown that return of synchrony is not immediate, but that it is important in the recovery process. The purpose of this study was to test a combination of cues that have been shown to adjust the timing of circadian temperature rhythm. The combined cues consisted of timed ingestion of caffeine and protein foods and adjustment of the sleep/wake cycle. The intervention was tested in 26 age- and gender-matched maxillofacial surgery patients. Patients were randomly assigned to control or experimental groups. Circadian temperature rhythm was measured by continuous monitoring with axillary probes and miniature recorders before and after surgery. Following surgery, both experimental and control subjects displayed 24-hour circadian temperature rhythms; however, the peak-to-trough difference was decreased more following surgery in the control subjects than in the subjects who had prepared for surgery by practicing the intervention. Control subjects also had less day-to-day stability in the phase of their rhythms following surgery. These results suggest that the intervention reduced circadian disruption following surgery and provides a way for patients to prepare themselves to resist rhythm changes. PMID- 11876466 TI - Effects of exercise conditioning on thermoregulatory response to anticholinesterase insecticide toxicity. AB - Chronic exercise conditioning has been shown to alter basal thermoregulatory processes (change in thermoregulatory set point) as well as the response to infectious fever Chlorpyrifos (CHP), an organophosphate insecticide, also affects thermoregulation, causing an acute period of hypothermia followed by a delayed fever. This study examined whether chronic exercise training in the rat alters the thermoregulatory response to CHP. Core temperature and motor activity were monitored by radiotelemetry in female Sprague-Dawley rats housed individually at an ambient temperature of 22 degrees C. The rats were either given continuous access to running wheels or housed in standard cages without wheels. The exercise group ran predominately at night. After 8 weeks, the rats were gavaged with corn oil or 15 mg/kg CHP. CHP induced a transient hypothermic response followed by a delayed fever, beginning 1 day after exposure. Relative to controls, T7 decreases were not significantly different between the exercise (1.6 degrees C) group and the sedentary (0.5 degrees C) group given CHP. The sedentary and exercise group administered CHP developed a fever the day after CHP treatment. The fever response was greater in the sedentary group and persisted for approximately 3 days post-injection. Fever of the exercise group persisted for just one-half of 1 day after CHP. It is well known that chronic exercise training improves aerobic capacity; however, trained rats were not protected from the hypothermic effects of CHP. Training did ameliorate the febrile effects of CHP. Thus, exercise training may afford protection to the toxic effects of organophosphate insecticides. PMID- 11876467 TI - Effects of hydration on febrile temperature patterns in rabbits. AB - Patients with fever have a predisposition to experience dehydration, which may alter their thermoregulatory responses to elevated body temperature. In view of the recent discovery of the antipyretic activity of arginine vasopressin (AVP), it is possible that dehydration has a beneficial role during fever. Dehydration may enhance endogenous antipyresis by stimulating AVP release, making aggressive fluid replacement, which may inhibit AVP release, undesirable during fever. This study addressed the effects of manipulation of hydration status on temperature and cardiovascular responses in endotoxin-injected rabbits. Eight unanesthetized chronically instrumented rabbits were exposed to lipopolysaccharide (LPS) while in euhydrated state, after furosemide (5 mg/kg) and 24 hours of water deprivation (dehydrated), after infusion of saline (30 mL/kg) while in euhydrated state (hyperhydrated), and after saline (mL/per overnight body weight loss in grams) while in dehydrated state (rehydrated). Dehydrated rabbits display higher fevers that are biphasic in nature and are accompanied by increased vasoconstriction and duration of mean arterial pressure increases, indicating that activation of antipyretic mechanisms in dehydrated rabbits was not sufficient to reduce body core temperature. In addition, fluid supplementation in euhydrated rabbits did not alter the febrile response; however, a marked decrease in heart rate was noted. Furthermore, fluid supplementation in dehydrated rabbits significantly attenuates the rectal temperature and heart rate response to LPS injection, indicating the possibility that activation of antipyretic mechanisms of AVP in rehydrated rabbits was sufficient to reduce body core temperature. The results suggest that fluid supplementation has a beneficial role in keeping body temperature lower . PMID- 11876468 TI - Quantitative reverse transcription-PCR measurement of tissue factor mRNA in glioma. AB - BACKGROUND: Initiation of the coagulation serine protease cascade in mammalian cells is mediated by tissue factor (TF), which is a cell surface receptor and cofactor for coagulation factor VII (FVII) and its activated form FVII (FVIIa). Increasing evidence suggests that TF is expressed in a wide range of cancer cells and plays important roles in cancer progression and metastasis. In this study, we investigated the association between the expression level of TF transcript and histologic features of glioma. METHODS: RNA was extracted from normal brain tissues and glioma tissues. We developed and validated a real-time quantitative reverse transcription (RT)-PCR assay, based on fluorescent TaqMan methodology, to quantify TF gene expression and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) at the mRNA level in human glioma. RESULTS: The dynamic range of the assay was 10(3)-10(8) copy/microg RNA. The relationship between Ct and log starting concentration was linear (r2 > or = 0.99). The mean expression of TF in healthy brain tissue was 6.2 x 10(3) copy/microg RNA. Overexpression of TF was found in 42 brain glioma samples, mean value is 2.9 x 10(6) copy/microg RNA. CONCLUSIONS: TF mRNA transcript is expressed in glioma and the level of expression correlates with histologic grade of malignancy. This new simple, rapid, semiautomated assay is a major alternative to Northern blot and competitive quantitative PCR for gene alteration analysis in human tumors and may be a powerful tool for large randomized, prospective cooperative group trials and support future TF-based clinical applications. PMID- 11876469 TI - Production of HIV-1 p24 protein in transgenic tobacco plants. AB - The production of antigens for vaccines in plants has the potential as a safe and cost-effective alternative to traditional production systems. Toward the development of a plant-based expression system for the production of human immunodeficiency virus type I (HIV-1) p24 capsid protein, the p24 gene was introduced into the genome of tobacco plants using Agrobacterium tumefaciens mediated gene transfer. Southern blot analyses confirmed the presence of the p24 coding sequence within the genome of transgenic lines. Western blot analysis of protein extracts from transgenic plants identified plant-expressed p24 protein that cross-reacted with a p24-specific monoclonal antibody, thus confirming the maintenance of antigenicity. Quantification of the p24 protein using enzyme linked immunosorbent assay (ELISA) estimated yields of approx 3.5 mg per g of soluble leaf protein. Similar accumulation levels of p24 were also detected in T1 plants, confirming that the p24 gene is transmitted stably. Our results indicate that plant-based transgenic expression represents a viable means of producing p24 for the development of HIV vaccine and for use in HIV diagnostic procedures. PMID- 11876470 TI - Isolation of differential genes in suspension cultures of Taxus cuspidata induced by additional taxol. AB - Addition of taxol into suspension cultures of Taxus cuspidata induced cell apoptosis, which was confirmed by gel electrophoresis of the DNA ladders indicating the progressive delineation of fragmented nuclear DNA (nDNA) into distinct bodies. The additional taxol not only changed the microtubule assembly of cells, but also affected the gene expression. Fourteen cDNA fragments, named as TIGT9-22, were isolated after addition of taxol and their GenBank accession numbers were given as BF704560-BF704573, respectively. Among them, TIGT13 and TIGT21 were apparently homogeneous with apbE and carbamoylphosphate synthetase, respectively. Other cDNA fragments showed no significant analogy with the known sequences in GenBank. PMID- 11876471 TI - Transfection of liver in vivo by biolistic particle delivery: its use in the investigation of cytochrome P450 gene regulation. AB - We describe the delivery of reporter gene constructs to rat liver through the use of the Helios Gene Gun system. The effectiveness of this transfection method is illustrated by describing its use for determining in vivo the role of a DNA element that regulates cytochrome P450 2B1 (CYP2B1) gene expression in response to xenobiotics. DNA was delivered to the liver of an anesthetized animal via DNA coated gold microcarriers. The highest level of reporter gene expression was obtained about six hours posttransfection; however, at this time endogenous CYP2B1 mRNA is transiently induced by the anesthetic treatment. The optimal time for investigating expression of a reporter gene under the control of CYP2B1 regulatory elements was 24 h after transfection, by which time the inductive effect of the anesthetic had ceased. Reporter gene expression subsequently declined rapidly to a low level by 48 h. In the transfected liver the heterologous SV40 promoter was about eight-fold stronger than the minimal CYP2B1 promoter. However, when attached to the phenobarbital response element both promoters give the same fold-induction of reporter activity in response to phenobarbital. PMID- 11876472 TI - Screening with tumor markers: critical issues. AB - Reviewing the literature it would appear that tumor markers have often flattered to deceive. Early promise does not often seem to be borne out in extended trials. Despite apparently high specificity, very few markers are capable of assisting in a screening process. This brief review attempts to put the roles of tumor markers in perspective and explain how their misapplication has led to misunderstanding of their potential value in a clinical context. It also considers the theoretical basis for their use and highlights how misunderstanding of these can lead to flawed studies and application. PMID- 11876474 TI - PCR-based methods for detecting DNA damage and its repair at the sub-gene and single nucleotide levels in cells. AB - Three PCR-based methods are described that allow covalent drug-DNA adducts, and their repair, to be studied at various levels of resolution from gene regions to the individual nucleotide level in single copy genes. A quantitative PCR (QPCR) method measures the total damage on both DNA strands in a gene region, usually between 300 and 3,000 base pairs in length. Strand-specific QPCR incorporates adaptations that allow damage to be measured in the same region as QPCR but in a strand-specific manner. Single-strand ligation PCR allows the detection of adduct formation at the level of single nucleotides, on individual strands, in a single copy gene in mammalian cells. If antibodies to the DNA adducts of interest are available, these can be used to capture and isolate adducted DNA for use in single-strand ligation PCR increasing the sensitivity of the assay. PMID- 11876473 TI - Characteristics and applications of nucleic acid sequence-based amplification (NASBA). AB - Nucleic acid sequence-based amplification (NASBA) is a sensitive, isothermal, transcription-based amplification system specifically designed for the detection of RNA targets. In some NASBA systems, DNA is also amplified though very inefficiently and only in the absence of the corresponding RNA target or in case of an excess (>1,000-fold) of target DNA over RNA. As NASBA is primer-dependent and amplicon detection is based on probe binding, primer and probe design rules are included. An overview of various target nucleic acids that have been amplified successfully using NASBA is presented. For the isolation of nucleic acids prior to NASBA, the "Boom" method, based on the denaturing properties of guanidine isothiocyanate and binding of nucleic acid to silica particles, is preferred. Currently, electro-chemiluminescence (ECL) is recommended for the detection of the amplicon at the end of amplification. In the near future, molecular beacons will be introduced enabling "real-time detection," i.e., amplicon detection during amplification. Quantitative HIV-1 NASBA and detection of up to 48 samples can then be performed in only 90 min. PMID- 11876475 TI - Direct analysis for familial adenomatous polyposis mutations. AB - The spectrum of disease causing mutations is immense. It just so happens that the overwhelming majority of genetic alterations in the APC gene with leads to adenomatous polyposis coli generate truncated gene products. This observation lead to the development of the in vitro synthesis protein assay (protein truncation test) which is a sensitive method to detect these truncated gene products from patient samples. This article describes the assay to detect truncated proteins for the APC gene, which can also be applied to other disease causing genetic alterations which commonly lead to truncations such in HNPCC, von Hippel-Lindau, osteogenesis imperfecta, retinoblastoma, BCRAI, beta-thalassemia, hemophilia B, Duchenene and Becker muscular dystrophy. PMID- 11876476 TI - Finding your knockout: reverse genetics techniques for plants. AB - The process of finding a mutant plant for your gene of interest has recently become far more straightforward and painless than has ever been possible before. This has come about through the production of large-scale insertional mutagenesis populations. These can now be readily screened in bulk for insertional mutants through a mixture of molecular (PCR/hybridization) and bioinformation techniques. Here we describe a step-by-step guide to the molecular protocols and a description of the bioinformatics approach. PMID- 11876478 TI - Cranial base kyphosis and the surface morphology of the anterior cranial fossa. AB - We investigated the relationship between the surface morphology of the anterior cranial fossa and cranial base kyphosis (sphenoid angle) in 52 cephalometric craniograms. Among them there were 25 female (mean age 54 +/- 15; range 31-82) and 27 male (mean age 43 +/- 18, range 19-85) skulls. The sphenoid angle and the altitudes of the highest elevation of the endofrontal eminence (cranial base over the orbital roof in the anterior cranial fossa) and the middle point of the sphenoid planum, measured according to the Frankfort horizontal, were analysed using classical cephalometric and morphometric analysis. Statistical analysis was performed by Pearson's product-moment correlation and simple linear regression. The sphenoid angle ranged from 97 degrees to 137 degrees (mean 118 +/- 9 degrees). The altitude ratio of the highest elevation of the endofrontal eminence and the middle point of the sphenoid planum ranged from 1.5 to 1.8 (mean 1.6 +/- 0.1). A significant correlation was found between this ratio and the sphenoid angle (r = -0.65; p < 0.001; coefficient of determination = 0.43). The elevation of the endofrontal eminence relative to the sphenoid planum was higher in skulls with increased cranial base kyphosis, whereas reduced sphenoid angle was associated with an increase in the elevations of the endofrontal eminence. Although the sphenoid angle has a significant effect on the morphology of the anterior cranial fossa, only 43% of the variance in altitude of the endofrontal eminence is likely to be explained by its relationship with the sphenoid angle. PMID- 11876477 TI - Aspects of the history of osteogenesis imperfecta (Vrolik's syndrome). AB - Osteogenesis imperfecta (OI) or Vrolik's syndrome is a heterogeneous group of inherited conditions arising from a variety of biochemical and morphological collagen defects. It was Willem Vrolik, Professor of Anatomy, Pathological Anatomy and Zoology at the Athenaeum Illustre (University of Amsterdam), who described in his Handbook of Pathological Anatomy (1842-1844) and Tabulae ad illustrandam embryogenesin hominis et mammalium, naturalem tam abnormem (1844 1849) a newborn infant with numerous fractures and hydrocephalus. In the Tabulae, having both Latin and Dutch texts, in the Latin text Vrolik used in the heading of Plate 91 the term Osteogenesis imperfecta (in Dutch: gebrekkige beenwording). Vrolik also mentioned that the infant lived three days and that both the parents were suffering from lues universalis at the time of birth. On our reexamination, the whole skeleton appeared poorly mineralised. The fairly large skull exhibited a broad and high forehead, large fontanels, frontal and temporal bossing, shallow orbits, and a protruding occiput. The calvaria consisted of many Wormian bones. The tubular bones, although of normal length and only slightly curved, were very thin, as were the ribs. All the skeletal structures showed one or more fractures and many fractures showed callus formation. In 1998 we re-diagnosed the condition of the specimen as osteogenesis imperfecta type II. Willem Vrolik was one of the first to realize that many skeletal dysplasias were not the result of a postnatal acquired disease, such as "rickets" or "osteomalacia" as many of his contemporaries believed. He thought that it might be due to insufficient intrinsic "generative energy." He substantiated this by stating that in this specimen a primary impairment of ossification is present and not a secondary degeneration. The descriptions given by Willem Vrolik in some of the specimens generated the term Osteogenesis imperfecta and the eponym Vrolik's syndrome for this genetic disorder characterized by increased fragility. PMID- 11876479 TI - The functional anatomy of the muscles of facial expression in humans with and without cleft lip and palate. A contribution to refine muscle reconstruction in primary cheilo- and rhinoplasties in patients with uni- and bilateral complete CLP. AB - The great variation of primary cheiloplasty procedures in Cleft Lip and Palate (CLP) patients shows that there is disagreement regarding the embryonic development of this part of the face, the macroscopic and microscopic functional anatomy of the human muscles of facial expression and their role as a functional matrix for balanced and harmonious facial development. The purpose of this study is to present results of microsurgically dissected facial muscles, several parts of the nose and the human midface in specimens with and without cleft lip and palate. The results are compared with those of other investigations. Recommendations are presented for a standardized dissection technique of the facial muscles of expression for different types of primary cheilo- and rhinoplasty techniques. PMID- 11876480 TI - Clinical implications of concomitant variations of the testicular, suprarenal and renal veins: a case report. AB - Multiple venous anomalies have been observed during dissection of the posterior abdominal wall in a 65-year-old, white male cadaver. The left testicular and suprarenal veins united inferior to the superior mesenteric artery, coursed anterior to the abdominal aorta and drained into the inferior vena cava (IVC). Further the left renal vein coursed retroaortically and divided into three branches. The superior branch coursed on the vertebral column and drained into the azygos vein while middle and inferior branches drained into the IVC. The right renal vein was double and both drained into the IVC separately. Due to implications for numerous therapeutical and diagnostic procedures in the retroperitoneal region knowledge of these variations could be useful for clinicians in its recognition and protection. PMID- 11876481 TI - Angiographic study of the arterial supply to the palatine mucoperiosteum. AB - Studies on the arterial supply of dog palatal mucoperiosteum have contributed to the success of surgical procedures on the hard palate of humans. We decided to undertake a more detailed anatomical study of the arterial supply of the canine palate. 15 male dogs of the species Canis familiaris were used. Contrast solution (barium sulfate) was injected into the palatal arterial system, followed by the removal of the dog's palate together with its mucoperiosteum. To obtain good radiographic imaging of the arterial network of the palatal mucoperiosteum, decalcification of the palatal bone was carried out. X-ray images were taken by a mammography X-ray machine set for 23 kV and 10 mA. The X-ray images showed that the mucoperiosteal arterial network is composed of a left and a right major artery which enters the mucoperiosteum through the left and right foramen, respectively. Anastomoses were observed between these two major arteries along their path from the posterior to the anterior regions of the mucoperiosteum. These anastomoses always occurred at the palatal transversal crests. The statistical study of the collected data showed that, for the posterior third of the palate, there is a proportionality between the number of the arterial derivations from the two major arteries and the mucoperiosteal area. The X-ray images also showed that the middle third of the palate is the least supplied with arterial derivations. PMID- 11876482 TI - Immunoelectron microscopic investigations of patching, capping, endocytotic and shedding processes in T and B lymphocytes. AB - Lymphocytes were isolated from the blood of healthy juvenile test persons by the FICOLL method. Subsequently, CD4-, CD8-, and CD19-positive cells were obtained by the use of magnetic beads. The sandwich technique and gold labelling method (preembedding) served for the demonstration of receptors in the electron microscope. The gold-labelled receptors were primarily endocytosed via smooth walled micropinocytotic processes, less frequently by coated pits/vesicles. The endocytosis cycle lasted only as far as the level of multivesiculated bodies. Lysosomes and structures of the Golgi apparatus were free from gold particles. It was surprising that after activation the capping phenomena were not associated with increased endocytotic activity. The inner surface of the membrane of endocytotic vesicles underneath the cap does not explain modulation or turnover of the receptor under these conditions, not even in view of a fast endocytotic cycle. Another possibility of a membrane turnover is the "shedding" process. We were indeed able to demonstrate gold labelling and surface coat-like material in the extracellular matrix. PMID- 11876483 TI - Phagocytosis of immunobeads by CD8 positive lymphocytes during magnetic cell sorting. AB - Lymphocytes of human blood were isolated by Ficoll gradients and separated into a CD8 positive fraction using the "Immunobead Technique". Electron microscopic examination approximately 45 min after mixing with the beads at room temperature revealed adhering beads in all cells of the CD8 positive fraction, whereas the negative fraction did not exhibit any adhering beads. About 2% of the lymphocytes of the positive fraction additionally showed bead incorporations. These cells exhibited a characteristic lymphocytic morphology and must be considered T lymphocytes. Formation of processes and ridges in the vicinity of the adhering beads points to a phagocytosis-like process. Since incorporations of beads can be demonstrated only in the CD8 positive fraction, this type of phagocytosis must be a specific; i. e. receptor-mediated phagocytotic process. The capability to perform phagocytosis by CD8 positive lymphocytes at room temperature should be considered during separation and when evaluating the behaviour of cells after magnetic separation, e. g. when attempting to remove the beads again. An exact experimental temperature (4 degrees C) should therefore be maintained. PMID- 11876484 TI - Glycoconjugate histochemistry of bovine Brunner glands. AB - The principal aims of this study have been to elucidate the nature of glycoconjugates produced by the two distinct parts of bovine Brunner glands, peripheral and central areas of lobules, and to investigate the presence of sialyl acid residues. Bovine duodenal tissues, embedded in paraffin wax, were investigated by means of both conventional histochemical methods (PAS, AB, HID) and biotinylated lectins (Con A, DBA, SBA, GS-I-B4, PNA, sWGA, GS-II, UEA-I, LPA, LFA). Conventional histochemical methods allowed us to accurately define two different areas: a central and a peripheral area. The central area, composed of secretory tubular tracts and the excretory duct, contained neutral glycoconjugates. The peripheral area was formed by both terminal alveolar and tubular secretory tracts and contained both neutral and acidic glycoconjugates, the latter partly carboxylated and partly sulfated. Lectin histochemistry confirmed differences highlighted by conventional histochemical methods and allowed us to characterise glycoprotein profiles of the preterminal and terminal tracts. The preterminal tracts and the excretory duct contained glycoconjugates with terminal D-Gal beta(1-3)GalNAc, alpha-D-Gal, alpha/beta-D-GalNAc, alpha/beta D-GlcNAc, and internal beta(1-4) D-GlcNAc and alpha-Man residues. The terminal tracts were characterised by terminal alpha-L fucose, beta-D-GalNac, alpha/betaD GlcNAc, alpha-D-Gal, alpha-D-GalNAc, and sialic acid residues. Internal beta(1-4) D-GlcNAc and alpha-Man residues were also identified. Finally, secretion of bovine Brunner glands is characterised by both O-linked and N-linked glycoproteins: cells located in the preterminal tracts and in the excretory duct produce mainly O-linked glycoproteins while cells located in the terminal tracts produce N-linked glycoproteins. PMID- 11876485 TI - Up-regulation of substance P and NMDA receptor mRNA in dorsal horn and preganglionic sympathetic neurons during adjuvant-induced noxious stimulation in rats. AB - Substance P (SP) and glutamate-containing terminals are found in the dorsal horn and preganglionic sympathetic neurons (PSNs) in the intermedio-lateral nucleus of the spinal cord. SP receptor (SPR) and N-methyl-D-aspartate type glutamate receptor (NMDAR) were also recognized in portions of the dorsal horn and PSNs. Primary sensory nerve fibers containing SP and glutamate terminated around PSNs, or partly on PSNs directly as well as on dorsal horn neurons (DHNs). The present study was performed to investigate the changes in SPR and NMDAR mRNA expressions during nociception in rats. Upon the injection of complete Freund's adjuvant (CFA) into the front paw, edema and hyperalgesia occurred immediately, with the difference in latency score between injected and non-injected paws continuing to day 10. The up-regulation of SPR and NMDAR mRNAs in DHNs and PSNs was recognized using in situ hybridization and northern blot techniques. CFA injection increased SPR mRNA expression in PSNs at days 1 and 4, and NMDAR mRNA expression at days 1, 4 and 7. At day 14, the mRNA expression of both receptors decreased to the control level. These changes in the amount of receptor mRNAs in DHNs and PSNs may cause hyperalgesia and sympathetically mediated pain. PMID- 11876486 TI - Route of lymphocyte migration through the high endothelial venule (HEV) in human palatine tonsil. AB - Eleven palatine tonsils were collected from subjects who underwent tonsillectomy in Christian Medical College Hospital and the route of migration of lymphocytes through the high endothelial vessel was studied under EM. In the interendothelial route, migration of a lymphocyte through HEV wall began with the adhesion of a lymphocyte to the surface of endothelial cells by means of a short cytoplasmic projection in the vicinity of intercellular space. The projection extended into the cleft between adjacent endothelial cells. The lymphocyte migrated through HEV by diapedesis. After the lymphocyte had traversed the interendothelial space, it occupied the subendothelial space. In the transendothelial route, migration of a lymphocyte through HEV was initiated by adherence of the lymphocyte to the endothelial cell. The adherent lymphocyte compressed or invaginated into the cytoplasm of the endothelial cell, entered the endothelial cell, was completely enclosed within the endothelial cell cytoplasm, and emerged from the endothelial cell to occupy the subendothelial space. Evidence is presented from static transmission electron microscopic pictures for the migration of lymphocytes by both interendothelial and transendothelial routes through the high endothelial venule. PMID- 11876487 TI - Macroscopical anatomy of the so-called "rotator interval". A cadaver study on 19 shoulder joints. AB - The triangular capsular space between the insertion tendons of the Mm. supraspinatus and subscapularis--the "rotator interval", can be divided into lateral, medio-superior and medio-inferior parts. The lateral part of the capsule is strengthened by the "Lig. semicirculare humeri" and the anterior fibres of the M. supraspinatus tendon. The Ligg. coracohumerale and "coracoglenoidale" are the macroscopical elements of the medio-superior part. The medio-inferior part of the "rotator interval" is reinforced by the Ligg. glenohumeralia superius et medium. The key ligament of the "rotator interval" is the "Lig. semicirculare humeri". Laterally it ensures the insertion of the anterior fibres of the M. supraspinatus tendon above the Lig. transversum humeri and on the Tubercula majus et minus. Medially it is the place of attachment of the Lig. coracohumerale and oblique fibres of the Lig. glenohumerale superius. The "rotator interval" is not a weak capsular region but a complex network of macroscopically recognizable tendinous and ligamentous structures. PMID- 11876488 TI - Types of neurons and some dendritic patterns of basolateral amygdala in humans--a golgi study. AB - Classification of the neurons in the human basolateral amygdala is performed on preparations impregnated by the Golgi technique. Three different neuronal types are found in the nuclei of the basolateral amygdala: Type I--Pyramidal cells, with numerous dendritic spines and two subtypes (slender and squat); Type II- Modified pyramidal cells, sparsely spinous with rare dendritic spines and two subtypes (single apical and double apical) and; Type III--Non-pyramidal cells, with few dendritic spines and three subtypes (bipolar, multipolar and gliaform). The analysis of the primary dendritic branches pointed out the occasional presence of dendritic bundles (fascicular dendritic arrangement) with their predomination in the parvicellular division of the basal nucleus and paralaminar nucleus. Additionally, the presence of dendrodendritic contacts, indicated by light microscopy, was also found in the parvicellular division of the basal nucleus and especially in the paralaminar nucleus. PMID- 11876489 TI - Vegetables, fruit, antioxidants and cancer: a review of Italian studies. AB - BACKGROUND: Case-control studies have suggested that a diet rich in fresh fruit and vegetables protects from the risk of most common epithelial cancers, including those of the digestive tract, and also several nondigestive neoplasms; however, selections in cohort studies have been generally weaker. AIM OF THE STUDY: To review the relation between frequency of consumption of vegetables and fruit, estimated intake of selected antioxidants and the risk of cancer at different sites. METHODS: Systematic overview of data, with specific focus on a network of case-control studies conducted in Italy from 1983 to 1999. RESULTS: The relative risks (RR) of digestive tract neoplasms were reduced in subjects reporting highest vegetable intake. A protective effect of vegetables was also observed for hormone-related neoplasms. Fruit was related to a reduced RR of cancers of the upper digestive tract, stomach and urinary tract. With reference to the role of selected antioxidants, beta-carotene, vitamins C and E showed a significant inverse relation with oral and pharyngeal, esophageal and breast cancer risk. Against colorectal cancer, the most consistent protective effects were provided by carotene, riboflavin and vitamin C, but inverse relations were observed also for calcium and vitamin D. CONCLUSIONS: Fruit and vegetable consumption in Mediterranean populations appears to provide protection against several types of neoplasms. PMID- 11876490 TI - Effects of dietary maritime pine seed oil on lipoprotein metabolism and atherosclerosis development in mice expressing human apolipoprotein B. AB - BACKGROUND: Conifer seeds are used for food preparation in several countries. Aim of the study To assess the lipid-lowering and antiatherogenic properties of maritime pine (Pinuspinaster) seed oil. METHODS: The effects of maritime pine oil supplementation (20% w/w) for 2 weeks were compared to those of coconut and sunflower oil in mice expressing human apolipoprotein B (hApoB). Atherosclerosis lesion development was measured in hApoB mice fed 1.25% (w/w) cholesterol and 0.05% (w/w) sodium cholate and either coconut, sunflower or maritime pine oil (20% w/w) for 8 weeks. RESULTS: After 2 weeks of dietary treatment, plasma cholesterol (p < 0.0001), triglyceride (p < 0.0003), phospholipid (p < 0.0001) and apolipoprotein B (p < 0.0001) levels were lower in mice supplemented with maritime pine oil than in those treated with coconut oil. These effects were accounted for by a lowering of LDL-cholesterol, LDL-phospholipids and LDL triglycerides, as well as a decrease in HDL-cholesterol and HDL-phospholipids. After 8 weeks of dietary treatment cholesterol and cholate, the mean area of aortic lesions was not statistically different between fat groups. CONCLUSIONS: Feeding maritime pine oil is associated with major changes of lipid and lipoprotein levels in hApoB mice. However, in the long term, maritime pine oil has no preventive effect on cholesterol-induced aortic lesion development in hApoB mice. PMID- 11876491 TI - Plasma levels of advanced glycation end products in healthy, long-term vegetarians and subjects on a western mixed diet. AB - BACKGROUND: Evidence indicates that food-derived Maillard's reaction products are absorbed and yet can be detected in the circulation. AIM OF THE STUDY: We postulated that consumption of the heat-treated food by omnivores could be reflected by higher plasma levels of advanced glycation end products (AGEs) in comparison with vegetarians, who in cooking (by keeping away from meat) use lower temperatures and less time for heating. METHODS: Plasma fluorescent AGEs (350/450 nm) and N(epsilon)-(carboxymethyl)lysine (CML, competitive ELISA) levels were investigated in 3 groups of healthy vegetarians (9 vegans-V, 19 lactoovo vegetarians--VLO and 14 semi-vegetarians--VS) and compared with those of age matched omnivores (O, n=19). Mean duration of vegetarian diet was V: 7.2 +/- 1.0,VLO: 8.2 +/- 0.8 and VS: 7.9 +/- 1.1 years. RESULTS: Both fluorescent AGE (O: 9.9 +/- 0.5; V: 10.8 +/- 0.7, LO: 13.1 +/- 0.8* and SV: 11.6 +/- 1.2 x 10(3) AU), and CML levels (O: 427.1 +/- 15.0,V: 514.8 +/- 24.6*, LO: 525.7 +/- 29.5**, SV: 492.6 +/- 18.0* ng/ml) were significantly lower in omnivores than in vegetarians. Plasma glucose, parameters of renal function (plasma concentration of creatinine and cystatin C, calculated glomerular filtration rate--GFR) as well as C-reactive protein levels were within the normal range and did not differ significantly between the groups. Thus, neither decline of kidney function nor inflammatory processes contributed to the rise in plasma AGEs. CONCLUSION: Enhanced plasma AGE levels in vegetarians in comparison to omnivores are herein presented for the first time. Mechanisms of AGE elevation and potential pathophysiological relevance of this finding are to be elucidated in prospective studies. PMID- 11876492 TI - Five year changes in waist circumference, body mass index and obesity in Augsburg, Germany. AB - AIMS: To assess temporal changes in body fat distribution, body mass index and obesity in Augsburg, Germany. METHODS: Waist circumference, weight and height were measured in two independent samples of 4804 and 4792, men and women, aged 25 74 years, in the MONICA Augsburg surveys 1989/90 and 1994/95. Abdominal obesity was defined as waist circumference greater than the 80th gender-specific percentile (men: 103, women: 92 cm) in the 1989/90 population. Obesity was defined as a body mass index (BMI) > or = 30 kg/m2. RESULTS: Age-standardized mean waist circumference increased by more than 1 cm (p-value < 0.00003) in both men and women while BMI increased by 0.3-0.4 kg/m2 (p-value < 0.01). We observed both a shift to higher values in the waist circumference distribution plus- particularly in women older than 45 years --a substantial right shift in the top of the distribution. Moreover, survey participants in 1994/95 who were at the higher end of the BMI distributions were disproportionately more obese than their respective peers in 1989/90. The prevalence of abdominal obesity rose by 3.3% in men and 3.6% in women, while the prevalence of obesity rose by 2% from 17% in men and by 2.5% from 19% in women. CONCLUSIONS: While changes in the Augsburg population may not be as alarming as in other countries, the secular increase in waist circumferences in both men and women occurring over a short time period indicates a need for prevention given the already high absolute weight, BMI and waist circumference levels in the population. PMID- 11876493 TI - Salicylic acid in soups prepared from organically and non-organically grown vegetables. AB - BACKGROUND: Salicylic acid is a chemical signal in plants infected by pathogens and it is responsible for the anti-inflammatory action of aspirin. Patients who take aspirin have a reduced risk of developing atherosclerosis and colorectal cancer, both of these pathologies having an inflammatory component. Dietary salicylic acid may help to prevent these conditions. We wondered if foods made from organically-reared plants might have a higher content of salicylic acid than those made from non-organic plants, since the latter are more likely to be protected from infection by the application of pesticides. OBJECTIVE: To determine if organic vegetable soups have a higher salicylic acid content than non-organic vegetable soups. METHODS: The contents of salicylic acid in organic and non-organic vegetable soups purchased from supermarkets were determined. Salicylic acid was identified by varying the chromatographic conditions and comparing the retention times of the unknown substance in the extracts with salicylic acid; by treating extracts of the soups with salicylate hydroxylase; and by using GCMS. Salicylic acid was determined by using HPLC with electrochemical detection. RESULTS: Salicylic acid was present in all of the organic and most of the non-organic vegetable soups. The median contents of salicylic acid in the organic and non-organic vegetable soups were 117 (range, 8 1040) ng x g(-1) and 20 (range, 0-248) ng x g(-1) respectively. The organic soups had a significantly higher content of salicylic acid (p=0.0032 Mann Whitney U test), with a median difference of 59 ng g(-1) (95 % confidence interval, 18 117ng x g(-1)). CONCLUSIONS: Organic vegetable soups contained more salicylic acid than non-organic ones, suggesting that the vegetables and plants used to prepare them contained greater amounts of the phenolic acid than the corresponding non-organic ingredients. Consumption of organic foods may result in a greater intake of salicylic acid. PMID- 11876494 TI - Effect of lactobacilli, bifidobacteria and inulin on the formation of aberrant crypt foci in rats. AB - BACKGROUND: Our studies were aimed at investigating the effect of lactic acid producing bacteria (LAB) or inulin, a natural source of non-digestible oligosaccharides derived from chicory, on the induction by carcinogens of aberrant crypt foci (ACF) in the colon, which are considered to be early precursor lesions of neoplasia. METHODS: Strains of Bifidobacterium longum, Lactobacillus casei and Lactobacillus acidophilus were administered to rats fed a purified high starch diet, under a variety of treatment protocols including daily gavage, via the drinking water and in the diet. The rats were treated with methyl N-nitrosourea, 1,2-dimethylhydrazine, or azoxymethane (AOM) to induce ACF. RESULTS: In general, no consistent significant changes in ACF numbers were detected in these experiments. In one study, the basal diet of the rats was changed to one containing a higher level of fat (corn oil). Under these conditions, a significant decrease in AOM-induced colonic ACF was seen in rats given L. acidophilus or inulin. In a concurrent group of animals fed a low fat diet, no significant decrease in ACF was observed. CONCLUSIONS: The results indicate that the type of diet fed can influence the detection of protective effects of LAB and oligosaccharides and that against the background of a diet with a level of fat typical of a Western diet, evidence for a protective effect of L. acidophilus and inulin towards colon cancer was obtained PMID- 11876495 TI - Health hazards due to the inhalation of amorphous silica. AB - Occupational exposure to crystalline silica dust is associated with an increased risk for pulmonary diseases such as silicosis, tuberculosis, chronic bronchitis, chronic obstructive pulmonary disease (COPD) and lung cancer. This review summarizes the current knowledge about the health effects of amorphous (non crystalline) forms of silica. The major problem in the assessment of health effects of amorphous silica is its contamination with crystalline silica. This applies particularly to well-documented pneumoconiosis among diatomaceous earth workers. Intentionally manufactured synthetic amorphous silicas are without contamination of crystalline silica. These synthetic forms may be classified as (1) wet process silica, (2) pyrogenic ("thermal" or "fumed") silica, and (3) chemically or physically modified silica. According to the different physicochemical properties, the major classes of synthetic amorphous silica are used in a variety of products, e.g. as fillers in the rubber industry, in tyre compounds, as free-flow and anti-caking agents in powder materials, and as liquid carriers, particularly in the manufacture of animal feed and agrochemicals; other uses are found in toothpaste additives, paints, silicon rubber, insulation material, liquid systems in coatings, adhesives, printing inks, plastisol car undercoats, and cosmetics. Animal inhalation studies with intentionally manufactured synthetic amorphous silica showed at least partially reversible inflammation, granuloma formation and emphysema, but no progressive fibrosis of the lungs. Epidemiological studies do not support the hypothesis that amorphous silicas have any relevant potential to induce fibrosis in workers with high occupational exposure to these substances, although one study disclosed four cases with silicosis among subjects exposed to apparently non-contaminated amorphous silica. Since the data have been limited, a risk of chronic bronchitis, COPD or emphysema cannot be excluded. There is no study that allows the classification of amorphous silica with regard to its carcinogenicity in humans. Further work is necessary in order to define the effects of amorphous silica on morbidity and mortality of workers with exposure to these substances. PMID- 11876496 TI - A repeated 28-day oral dose toxicity study of 17alpha-methyltestosterone in rats, based on the 'enhanced OECD Test Guideline 407' for screening the endocrine disrupting chemicals. AB - As part of the international validation project to establish the Enhanced OECD Test Guideline 407, we performed a 28-day repeated-dose toxicity study of 17alpha methyltestosterone, an exogenous androgen agonist. Special attention was paid to the sensitivity of additional parameters for detecting endocrine-related effects of endocrine-disrupting chemicals, based on the existing Test Guideline 407. Seven-week-old Crj:CD(SD)IGS rats were allocated to one of four groups, each consisting of ten males and ten females, and 17alpha-methyltestosterone was administered daily by gavage at doses of 0 (control), 5, 20 and 80 mg/kg body weight per day. Male rats were killed on the day after the 28th administration and females on the day of the diestrus stage during the 4 day period after the 28th administration. Male rats receiving 80 mg/kg 17alpha-methyltestosterone demonstrated decreases in testis and epididymis weights, atrophy of seminiferous tubules and Leydig cells, and degenerated pachytene spermatocytes in the testes and degenerated germ cells in the epididymides as major alterations. Female rats showed abnormal estrous cycles, decreases in ovary and adrenal weights, increase in immature follicles with decreased corpus lutea in the ovaries at doses of 5 mg/kg and higher, as well as atrophy of zona reticularis in the adrenals and increase in mammary gland secretion at 20 mg/kg and above. Dilatation of the lumina and apoptosis of endometrial cells in the uterus, mucinification in the vagina and increase in serum follicle-stimulating hormone were seen with 80 mg/kg. Among the parameters examined in the present experimental system, effects of 17alpha-methyltestosterone on endocrine-related organs were detected in organ weights and histopathological examination of both sexes, and in serum hormones and estrous cycle of females. Based on these results, the no-observed-adverse effect level (NOAEL) in the present study was estimated to be below 5 mg/kg per day. In particular, effects were most sensitively detected by organ weights and histopathological examination of sexual organs. PMID- 11876497 TI - Toxicokinetic interaction of 2,5-hexanedione and methyl ethyl ketone. AB - Co-exposure to methyl ethyl ketone (MEK) potentiates the neurotoxicity of n hexane in humans as well as in animals. This effect is associated with increased persistence of 2,5-hexanedione (2,5-HD) in blood, probably due to inhibition of 2,5-HD phase II biotransformation by MEK. There is no previous quantitative toxicokinetic model to describe this interaction. In this study we constructed a toxicokinetic model to depict the inhibition of 2,5-HD metabolism and elimination by MEK. Experimental data on 2,5-HD blood concentrations in rats from a published study were used to estimate model parameters. Three different inhibition mechanisms were evaluated: competitive, uncompetitive, and noncompetitive inhibition. Extrapolation from high to low doses was made to assess the interactive effects of MEK on 2,5-HD beyond experimental conditions. The models developed successfully described the toxicokinetic behavior of 2,5-HD when inhibited by MEK. The competitive inhibition model yielded a much lower estimate for the constant (65.5 mg/l) of 2,5-HD inhibition by MEK than did the uncompetitive and noncompetitive models (403 and 440 mg/l, respectively). The apparent half-life of 2,5-HD appeared to be a linear function of the Michaelis Menten constant, and 2,5-HD and MEK concentrations in rats. The area under the curve of 2,5-HD in blood of rats was a nonlinear function of 2,5-HD and MEK concentrations in the blood. This study highlights the importance of the interactive effect of MEK on deactivation and elimination of 2,5-HD, and further illustrates the advantage of toxicokinetic modeling to investigate chemical interactions associated with exposure to multiple chemical agents. PMID- 11876498 TI - Tissue concentrations and induction of a hepatic monooxygenase in male Wistar rats after repeated doses of defined polychlorinated dibenzo-p-dioxin and dibenzofuran (PCDDs and PCDFs) mixtures. AB - Two groups of male Wistar rats were treated 16 times (every 3rd day) subcutaneously with a defined mixture of polychlorinated dibenzo-p-dioxins (PCDDs) or of polychlorinated dibenzofurans (PCDFs). These mixtures contained no measurable amount of 2,3,7,8-TCDD. Each single dose was calculated to contain either 57 ng I-TEq (international 2,3,7,8-T4CDD toxicity equivalencies)/kg body weight of the PCDD mixture or 39 ng I-TEq/kg body weight of the PCDF mixture. Both mixtures contained a large excess of non-2,3,7,8-substituted congeners. The activities of ethoxyresorufin O-deethylase (EROD) in liver microsomes were correlated with the corresponding concentrations of PCDDs or PCDFs in hepatic tissue. Data were compared with results obtained after single injections of 2,3,7,8-tetrachlorodibenzo-p-dioxin (2,3,7,8-T4CDD). As expected, a complex kinetic situation resulted, because of the different tissue distributions and elimination half-lives of the various congeners: (1) 2,3,7,8-substituted PCDDs: the time course of the concentrations in liver and adipose tissue was similar for all congeners, the levels increased during the treatment period and decreased after treatment. Tissue concentrations of all 2,3,7,8-substituted PCDDs were considerably higher in liver than in adipose tissue. The liver/adipose tissue concentration ratios increased with the degree of chlorination. The ratio of 1,2,3,7,8-P5CDD was much lower than those of all other 2,3,7,8-substituted congeners. (2) 2,3,7,8-substituted PCDFs: 1,2,3,7,8-P5CDF was rapidly eliminated from liver and adipose tissue while 2,3,4,7,8-P5CDF largely persisted after the treatment period in both tissues. 2,3,7,8-T4CDF was eliminated even more rapidly than 1,2,3,7,8-P5CDF and could not be detected after treatment in both tissues. Time courses of the concentrations of 2,3,4,7,8-P5CDF, H6CDFs, H7CDFs and OCDF in liver and adipose tissue were similar: the levels of all congeners increased during the treatment period but no clear-cut decrease was observed within 34 days after the last treatment. Tissue concentrations of all 2,3,7,8-substituted PCDFs were higher in liver than in adipose tissue. The liver/adipose tissue concentration ratios increased with the degree of chlorination. The ratios of 2,3,7,8-T4CDF and 1,2,3,7,8-P5CDF were much lower than those of all other 2,3,7,8 substituted congeners. (3) non-2,3,7,8-substituted PCDDs and PCDFs: a number of non-2,3,7,8-substituted PCDD and PCDF congeners were found in both tissues in concentrations below 1 ng/g. In adipose tissue nearly all congeners were found during the treatment period showing a decrease after the treatment. In liver samples, many higher chlorinated PCDF congeners (with >4 chlorine atoms) could be detected. Most of those substituted in three of the four 2, 3, 7 and 8-positions persisted after treatment. In contrast, only one 1,4,6,9-substituted isomer of each PCDD homologue group was found during treatment with high recoveries after the third injection, but a rapid decline occurred already during the treatment period. (4) EROD activity: a good linear relationship (when using a double-log plot) between the EROD activities and the hepatic concentrations (ng I-TEq/g tissue) was found both in the PCDD-treated (r2= 85.8%) and in the PCDF-treated group (r2=87.3%). A similar correlation (r2=95.6%) was observed in rats treated with 2,3,7,8-TCDD alone (concentration range in liver tissue: 0.2 to 9.7 ng/g wet weight). The concentration-response curves for both the PCDD and PCDF mixtures run parallel to the curve for 2,3,7,8-T4CDD. However, the inductive potency of the PCDD or PCDF mixture was approximately 3-fold or 4-fold lower, respectively, compared with the inductive potency of 2,3,7,8-T4CDD. Thus, the I-TE factors overestimated the potency of the mixtures in the concentration range tested and taking EROD induction as an end point. PMID- 11876499 TI - Metabolism and CYP-inducer properties of astaxanthin in man and primary human hepatocytes. AB - Previous investigations in the rat have shown that the non-provitamin A carotenoid astaxanthin is metabolized into 3-hydroxy-4-oxo-beta-ionone and 3 hydroxy-4-oxo-7,8-dihydro-beta-ionone and, in addition, is a potent CYP1A gene inducer. Here we investigated the metabolism of this compound as well as its capacity to induce CYP genes in primary cultures of human hepatocytes. Free metabolites of 14C-astaxanthin produced in this cellular model were purified by high pressure liquid chromatography (HPLC) and identified by gas chromatography mass spectrometry (GC-MS) analyses as 3-hydroxy-4-oxo-beta-ionol and 3-hydroxy-4 oxo-beta-ionone. In addition, deconjugation of polar compounds by glusulase and further analyses with HPLC and GC-MS revealed four radiolabeled metabolites including: 3-hydroxy-4-oxo-beta-ionol, 3-hydroxy-4-oxo-beta-ionone, and their reduced forms, 3-hydroxy-4-oxo-7, 8-dihydro-beta-ionol and 3-hydroxy-4-oxo-7,8 dihydro-beta-ionone. The same four metabolites were identified in human plasma from two volunteers who had orally taken 100 mg astaxanthin 24 h before blood collection. In cultured hepatocytes, astaxanthin was a significant inducer of the major cytochrome P450 enzyme, CYP3A4 as well as of CYP2B6, but not of other CYPs, including those from CYP1A and CYP2C families. The lack of autoinduction of astaxanthin metabolism in human hepatocytes suggests that neither CYP3A4 nor CYP2B6 contribute to the formation of metabolites. We conclude that metabolism of astaxanthin and its CYP-inducing capacity are different in humans and in rats. The novel methodology used in our studies could be extended to evaluating the role of metabolites of more important carotenoids such as beta-carotene in differentiation and carcinogenicity. PMID- 11876500 TI - Age-related effects of chlorpyrifos on muscarinic receptor-mediated signaling in rat cortex. AB - Chlorpyrifos (CPF) is a widely used organophosphorus pesticide. Earlier work from our laboratory and others has demonstrated that the sensitivity to CPF exposure changes markedly during maturation. A number of studies suggest that in addition to inhibiting acetylcholinesterase (AChE), CPF oxon may also interact directly with m2 and/or m4 subtypes of muscarinic acetylcholine receptors (mAChRs). In the present study, we investigated the in vivo effects of CPF exposure on phosphoinositide (PI) hydrolysis and cAMP formation, second-messenger systems coupled to m1, m3 and m5 (PI hydrolysis) or m2 and m4 (cAMP formation) mAChRs. Neonatal (7-day), juvenile (21-day) and adult (90-day) rats were treated with either peanut oil s.c. or CPF s.c. at 0.3x or 1x the maximum tolerated dosage (MTD: 45, 127 and 279 mg/kg for 7-day, 21-day and 90-day rats, respectively). Neurochemical end-points including AChE activity, muscarinic receptor ([3H]quinuclidinyl benzilate, and [3H]oxotremorine) binding, PI hydrolysis, and cAMP formation in cortex were evaluated at 4 h, 24 h, or 96 h after treatment. Under these conditions, relatively similar maximal degrees of cholinesterase (ChE) inhibition were noted, but times to peak inhibition varied among these age groups (24 h in neonates and juveniles, 96 h in adults). Total muscarinic receptor (QNB) binding was reduced in all three age groups with 1x MTD exposure, at both 24 h and 96 h in neonates and juveniles, but only at 96 h in adults. Oxotremorine binding was also reduced at 96 h after MTD exposure in all three age groups. Neither basal nor carbachol-stimulated IP accumulation was affected in any age group or at any time point following CPF exposure. In contrast, basal cAMP formation was significantly increased by MTD exposure in all three age groups 4 h after exposure, and at 4 h, 24 h, and 96 h after exposure in juveniles. Forskolin/Mn2+-stimulated cAMP formation was increased in neonates and juveniles at 96 h, and in juveniles also at 24 h, but was significantly decreased in adults at 96 h after MTD exposure. Oxotremorine-mediated inhibition of cAMP formation was significantly greater at 96 h after MTD exposure in all three age groups. These results provide further evidence that the cortical cAMP signaling pathway may be particularly sensitive to CPF exposure in neonatal, juvenile, and adult rats, possibly due to a direct interaction between CPF (or its oxon) and mAChRs or other components of the adenylyl cyclase cascade. PMID- 11876501 TI - The coffee components kahweol and cafestol induce gamma-glutamylcysteine synthetase, the rate limiting enzyme of chemoprotective glutathione synthesis, in several organs of the rat. AB - The coffee components kahweol and cafestol (K/C) were reported to be protective against mutagenic damage by heterocylic amines and aflatoxin B1 in the rat, while in humans the consumption of coffee with a high K/C content was associated with a lower rate of colon tumors. An important mechanism of this antimutagenic effect appears to be the potential of K/C to induce glutathione-S-transferase (GST) and to enhance hepatic levels of glutathione (GSH), the co-factor of GST, which is independently involved in further protective mechanisms. In the present study, we investigated mechanisms and organ specificities (liver, kidney, lung, colon) of the K/C effect on GSH levels, and particularly the role of gamma-glutamylcysteine synthetase (GCS), the rate limiting enzyme of GSH synthesis. Chows containing one of four concentrations of either a 1:1 mixture of K/C (0.012-0.122%) or of cafestol alone (0.006-0.061%) were fed to male F344 rats for 10 days. In the K/C treated livers, a dose-dependent increase of up to 2.4-fold in the activity of GCS was observed, being statistically significant even at the lowest dose, and associated with an increase in GSH of up to three-fold. Notably, the highest dose doubled the hepatic mRNAs of the heavy and light subunits of GCS, suggesting enhanced transcription. In the extrahepatic organs, GCS activity and GSH levels were increased as well, although more moderately than in the liver. Since enhancement of GCS had also been observed as a consequence of oxidative stress, the possibility of such an involvement in the actions of K/C was examined by determining hepatic thiobarbituric acid reactive substances and the ratio of oxidized and reduced GSH. However, no evidence of oxidative stress was detected. In summary, K/C increased GSH levels apparently through the induction of the rate limiting enzyme of GSH synthesis, which may be a key factor in the chemopreventive potential of coffee components. PMID- 11876502 TI - Novel action of lignans isolated from Hernandia nymphaeifolia on Ca2+ signaling in human neutrophils. AB - The effects of five lignans (epi-aschantin, epi-magnolin, epi-yangambin, deoxypodophyllotoxin, yatein) isolated from Hernandia nymphaeifolia (Presl.) Kubitzki (Hernandiaceae) on intracellular Ca2+ levels ([Ca2+]i) in human neutrophils were investigated by using fura-2 as a fluorescent probe. In both Ca2+-containing and Ca2+-free media, the lignans (50-100 microM) did not alter basal [Ca2+]i but inhibited the [Ca2+]i increase induced by platelet activating factor (PAF, 10 microM), leukotriene B4 (LTB4, 0.2 microM), and thapsigargin (1 microM) to different extents. In Ca2+-free medium, after depleting stores of Ca2+ with PAF, LTB4 or thapsigargin, addition of 3 mM Ca2+ induced Ca2+ influx. Each of the lignans (50-100 microM) caused 39-89% inhibition of PAF-induced Ca2+ influx; whereas only epi-aschantin was able to inhibit LTB4- and thapsigargin induced Ca2+ influx by 54-79%. Together, the results suggest that in human neutrophils, these lignans did not alter basal [Ca2+]i but inhibited Ca2+ movement induced by Ca2+ mobilizing agents. PMID- 11876503 TI - Uterotrophic and Hershberger assays for n-butylbenzene in rats. AB - We performed the uterotrophic and Hershberger assays proposed by the OECD to investigate the estrogenic and androgenic effects of n-butylbenzene (nBB). For the uterotrophic assay, nBB was injected subcutaneously at doses of 0, 40, 200, 1000 and 2000 mg/kg to 19-day-old rats for 3 days. In some rats, ethynylestradiol (EE) was also injected subcutaneously at a dose of 0.6 microg/kg after the administration of nBB. There were essentially no differences in the uterine wet or blotted weights between the control and any of the nBB-treated groups, or between the control and any of the nBB plus EE-treated groups. For the Hershberger assay, nBB was administered orally at doses of 0, 200 and 600 mg/kg to 56-day-old castrated rats for 10 days. In some rats, 0.4 mg/kg testosterone propionate (TP) was also administered by subcutaneous injection after the administration of nBB. Doses of 0, 200 and 600 mg/kg nBB were also administered orally to non-castrated rats. The weights of the accessory sex organs of the castrated rats showed no significant differences between the control and any of the nBB-treated groups or between the control and the nBB plus TP-treated groups. No significant differences in the weights of the accessory sex organs of the non castrated rats were observed between the control and the nBB-treated groups. These findings suggest that nBB does not have any endocrine-disrupting properties on in vivo screening tests. PMID- 11876504 TI - Steroid hormone progesterone induces cell proliferation and abnormal mitotic processes in rat liver. AB - The aim of this study was to evaluate the acute hepatic effects exerted by the steroid hormone progesterone (PR) in the rat. Although the liver is not a target tissue for this hormone, a number of hepatic actions of PR have been described, and, furthermore, a specific binding site for PR (PBS) exists in rat liver microsomes. Immature male rats were treated intraperitoneally with 60 mg/kg PR per day for 1, 5 or 10 days, and different parameters were evaluated in order to detect possible alterations in liver cells. Morphological study of the livers did not present images of cytotoxicity in any group of animals. The presence of a clear hyperplasia of smooth endoplasmic reticulum (SER) was noteworthy, mainly seen in perilobular hepatocytes. Despite this SER increase, the levels of cytochrome P450 (Cyt P450) significantly decreased after 10 days of PR administration. Similarly, the concentration of PBS was significantly decreased after 10 days of treatment with PR. On the other hand, these studies revealed a clear increase of mitotic activity and Ki-67 labelling index in the livers of animals treated with PR; furthermore, livers of PR-treated animals showed an increased percentage of binucleate hepatocytes. Flow cytometry analysis showed that although ploidy status of liver cells was not modified in any case the percentage of diploid nuclei in S-phase decreased during treatment with PR. The most relevant finding was the presence of abnormal mitosis and c-mitosis in livers from animals from all PR-treated groups. This study demonstrates that PR (a) does not induce cytotoxicity although it can induce cell proliferation and spindle disturbances in liver cells, (b) may also modulate the drug-metabolizing liver enzyme function, and (c) downregulates the expression of its own microsomal specific binding site. PMID- 11876505 TI - Embryonic exposure to lead: comparison of immune and cellular responses in unchallenged and virally stressed chickens. AB - Lead, a ubiquitous environmental contaminant, has been shown to modulate various functions of the immune system and decrease host resistance to infectious disease. However, limited information is available concerning the direct effects of lead on the host immune response to an infectious agent after developmental exposure. The current study utilized chickens to examine the effect of embryonic lead exposure on immune and cellular responses during viral challenge. Sublethal doses of lead were introduced into fertilized Cornell K Strain White Leghorn chicken eggs via the air sac at day 5 or day 12 of embryonic development (designated as E5 and E12, respectively). Four-week-old female chickens were inoculated with infectious bronchitis virus (IBV) strain M41. Antibody titer to IBV, delayed-type hypersensitivity (DTH) response against bovine serum albumin (BSA), the absolute number and percentage of leukocyte subpopulations, and interferon-gamma (IFN-gamma)-like cytokine production by splenocytes were evaluated at 5-6 weeks of age. While antibody response to IBV in juvenile chicks was unaffected by the in ovo lead exposure, IFN-gamma-like cytokine production by splenocytes was significantly depressed following lead exposure at both developmental stages. In contrast with this pattern, the DTH response against BSA was unaffected following E5 exposure, but was significantly decreased after E12 exposure to lead. These changes were similar to those previously reported in chickens not exposed to IBV. While lead exposure at E5 induced significant changes in the percentage of circulating heterophils at 1 day postinfection (dpi), lead did not cause any change in relative leukocyte counts after E12 exposure. At 7 dpi, E5 lead exposure resulted in decreased absolute number and percentage of circulating lymphocytes, while total leukocyte counts, and the absolute number and percentage of circulating monocytes and heterophils were significantly reduced in E12 lead-exposed chickens. These results suggest that low-level exposure to lead has a direct effect on the developing chicken immune system, which is evident even during a postnatal infection. Furthermore, some of the changes were observed only when chicks were stressed by the viral infection. It appears that lead exposure during different stages of embryonic development is likely to result in different immunotoxic outcomes in juveniles. PMID- 11876506 TI - Ultrastructural characterization of murine limb buds after in vitro exposure to grepafloxacin and other fluoroquinolones. AB - The effects of selected quinolones (levofloxacin, lomefloxacin, temafloxacin and grepafloxacin) on growth and differentiation of murine limb buds were studied in vitro. Ciprofloxacin and ofloxacin served as controls. We used limb buds from 12 day-old mouse embryos that were grown for 6 days in a serum-free, standard or magnesium-deficient medium. Besides evaluation under a dissecting microscope, we used electron microscopy to characterize the effects in detail. The following results are noteworthy. (1) Comparing the effects of standard and magnesium deficient medium after 3 and 6 days in culture, we found ultrastructural changes after 6 days only. (2) Direct comparison of ofloxacin (racemate) and levofloxacin (L-enantiomer) showed that they had a similar, rather low, potential for affecting cartilage development. (3) The effects of temafloxacin and ciprofloxacin were more pronounced in magnesium-deficient medium, but those of the other drugs were not. (4) Grepafloxacin was the most active quinolone in this assay. It impaired growth and differentiation of limb buds at 30 mg/l; at higher concentrations the explants did not grow. With lower concentrations of 10 mg grepafloxacin/l, no effects were detectable under a dissecting microscope but characteristic changes were seen by electron microscopy. We observed electron dense aggregates on and within chondrocytes, detachment of the cell membrane from the matrix with matrix-free pericellular areas around chondrocytes, and swelling of cell organelles such as mitochondria and rough endoplasmic reticulum. (5) The affinity of grepafloxacin for divalent cations (Mg2+, Ca2+) was studied by measuring the fluorescence of grepafloxacin solution at various concentrations of Mg2+ and Ca2+. Grepafloxacin showed a relatively high affinity for Ca2+ in the fluorescence assay, which was more pronounced than the affinities of six other fluoroquinolones tested before. PMID- 11876507 TI - Detection of ochratoxin A-induced DNA damage in MDCK cells by alkaline single cell gel electrophoresis (comet assay). AB - The mycotoxin ochratoxin A (OTA), a widespread contaminant of food and feedstuffs, is nephrotoxic, immunosuppressive and carcinogenic in domestic and laboratory animals. Additionally, it is suspected as being responsible for urinary tract tumours in patients suffering from Balkan endemic nephropathy. Moreover, evidence has accumulated that OTA is a genotoxic carcinogen, although the mechanism that results in DNA damage has not been fully resolved. In this study, the induction of DNA damage by OTA and the subsequent DNA repair was investigated by alkaline single cell gel electrophoresis (comet assay) in cells originally derived from the kidney, a target organ of OTA. With modifications of the method, the influence of OTA uptake into the cells and of DNA repair on the genotoxic effect of OTA should be investigated. In Madin-Darby canine kidney (MDCK) cells, OTA induced single-strand breaks in a concentration dependent manner. When an external metabolising enzyme system (S9-mix from rat liver) was added, this genotoxic effect was significantly stronger. By co-incubation with methotrexate or with the mycotoxin citrinin, a substrate of the organic anion transporter, the adverse effect of OTA was inhibited. When DNA repair was inhibited by addition of cytosine arabinoside and hydroxyurea, the tail length increased dramatically and all treated cells showed single-strand breaks. A further culture of the damaged cells in the absence of any supplement resulted in a complete repair of the DNA damage within 2 h. Adverse effects on the mechanisms of DNA repair, or exposure to OTA in periods of reduced DNA repair capacity may influence the genotoxic potency of OTA and have to be regarded as a further mechanism by which genotoxic effects of OTA can be performed. PMID- 11876508 TI - Vectorborne infectious diseases. PMID- 11876509 TI - Therapeutic drug monitoring of indinavir in HIV-infected patients undergoing HAART. AB - BACKGROUND: Therapeutic drug monitoring (TDM) of protease inhibitors (PI) is gaining increasing importance for the management of HIV-infected patients undergoing highly active antiretroviral therapy (HAART). The PI indinavir (IDV) is widely used in HAART regimens. Combinations of IDV with ritonavir (RTV) have been used to increase the plasma concentration of IDV. However, the desirable IDV concentration range in clinical practice remains to be elucidated. PATIENTS AND METHODS: To study the value of TDM for IDV in clinical practice, a retrospective analysis of 501 plasma samples of patients treated with IDV in various dosages was performed. IDV levels were determined during routine outpatient visits. Analysis was performed by high pressure liquid chromatography (HPlC) with UV detection. RESULTS: A widespread range of IDV plasma concentrations was seen both within and between patients. The mean IDV level during therapy with IDV 2.4 g/d was 3,260 ng/ml (95% CI: 2,903 ng/ml; 3,618 ng/ml). IDV levels at a dose of IDV 1.6 g/d in combination with RTV resulted in a mean IDV plasma concentration of 4,191 ng/ml (95% CI: 3,356 ng/ml; 5,026 ng/ml). There was no significant difference between plasma levels at the doses of 2.4 g/d and 1.6 g/d. 35 of all 130 patients treated with IDV reached only suboptimal IDV plasma concentrations below the limit of 150 ng/ml. There was no statistically significant difference between the number of patients below an IDV plasma concentration of 150 ng/ml in the various dosage regimens. CONCLUSION: During therapy with IDV in a b.i.d. scheme, similar IDV plasma concentrations and a comparable number of patients with subinhibitory plasma concentrations were observed when compared to a therapeutic regimen with t.i.d. dosing. In this study, even at various times of plasma sampling after oral ingestion, TCM facilitated the surveillance of patients compliance. PMID- 11876510 TI - Short courses of intravenous empirical antibiotic treatment in selected febrile neutropenic children with cancer. AB - BACKGROUND: Since the optimal duration of antibiotic therapy in febrile neutropenic patients is not clear, we evaluated the safety and efficacy of short courses of intravenous antibiotic treatment in selected pediatric cancer patients admitted for fever and neutropenia. PATIENTS AND METHODS: We retrospectively analyzed the clinical course of children with chemotherapy-induced neutropenia and fever. All patients were treated with empirical intravenous antibiotics. In episodes of fever of unknown origin (FUO), treatment regimen allowed discontinuation of antibiotics and early hospital discharge regardless of absolute neutrophil count (ANC) or evidence of bone marrow recovery as long as patients were afebrile for at least 24 h and had been treated for a minimum of 72 h. RESULTS: 106 episodes of febrile neutropenia occurred in 56 patients. 84 episodes were classified as FUO and intravenous antibiotic therapy was discontinued regardless of ANC when patients met the criteria described above. No death or major complication occurred. None of the patients had to be rehospitalized for recurrent fever or infection. CONCLUSION: Discontinuation of intravenous antibiotics regardless of ANC or evidence of bone marrow recovery seems safe and effective in pediatric cancer patients with FUO when children are afebrile for at least 24 h and are treated for a minimum of 72 h. PMID- 11876511 TI - Use of fibrates in the management of hyperlipidemia in HIV-infected patients receiving HAART. AB - BACKGROUND: Despite potent antiretroviral activity, protease inhibitor-based pharmacological treatment of HIV disease has recently been associated with lipid and glucose metabolism abnormalities (more frequently hypertriglyceridemia and hypercholesterolemia). The aim of our open-label, randomized, prospective study was to evaluate the role of fibrates in the management of HIV-associated hyperlipidemia. PATIENTS AND METHODS: Plasma lipid levels of 635 HIV-infected patients referred to our tertiary care center and who had been receiving protease inhibitor-based antiretroviral therapy for at least 12 months were evaluated. All patients presenting hypertriglyceridemia (> 300 mg/dl) of at least 6-month duration and unresponsive to a hypolipidemic diet and physical exercise were treated with bezafibrate (400 mg once daily), gemfibrozil (600 mg twice daily) or fenofibrate (200 mg once daily) for 12 months. RESULTS: 69 (10.9%) of the 635 observed patients received fibrate therapy: bezafibrate was employed in 25 cases, gemfibrozil in 22 and fenofibrate in 22. Hypolipidemic drugs led to a reduction of 41.2% and 23.3% vs baseline triglyceridemia and cholesterolemia, respectively, with a favorable tolerability profile. CONCLUSION: All used fibrates showed a similar, significant efficacy in the treatment of diet-resistant hyperlipidemia, but further studies seem necessary in order to establish the most appropriate guidelines for the management of dyslipidemia associated with highly active antiretroviral therapy (HAART). PMID- 11876512 TI - Norwalk virus gastroenteritis among Israeli soldiers: lack of evidence for flyborne transmission. AB - BACKGROUND: Paired sera collected from subjects before and after a fly-control intervention trial conducted in the Israel Defense Force (IDF) were tested for seroconversion to Norwalk virus (NV) to examine the role of NV as a cause of diarrhea in this population and to ascertain whether flies might also be implicated in transmission. MATERIALS AND METHODS: An enzyme-linked immunosorbent assay (ELISA), using recombinant NV capsid proteins (rNV) as antigen was employed to determine the seroconversion rate in a sample of 444 subjects. RESULTS: During 11-week field training cycles, 18% of IDF soldiers who were tested had an NV infection defined as a > or = 4-fold rise in antibody, yielding a cumulative incidence of nearly one infection (0.95) per soldier per year. The rate of seroconversion was nearly twice as high among soldiers who recalled having diarrhea as among those who did not, but the rates did not differ significantly between soldiers in the fly intervention areas and those in the control areas. CONCLUSION: NV is a common cause of enteric infections and diarrhea among Israeli soldiers who serve under field conditions, but unlike infections with Shigella and enterotoxigenic Escherichia coli, transmission of NV cannot be interrupted with an aggressive program of fly-control. PMID- 11876513 TI - Interleukin-3 and interleukin-4 in childhood brucellosis. AB - BACKGROUND: The Th1/Th2 cytokine balance seems to be involved in the susceptibility or resistance to Brucella infection, however the precise role of interleukins in human brucellosis has not been thoroughly explored. PATIENTS AND METHODS: The serum levels of interleukin-3 (IL-3) and interleukin-4 (IL-4) were measured by enzyme-linked immunosorbent assay (Quantikines, R&D Systems) in five children hospitalized for brucellosis and in a control group comprising ten children who were residents of the same area. RESULT: IL-4 levels were significantly increased during both the acute phase (p = 0.0036) and convalescence (p = 0.026) as compared to controls. IL-3 levels were mildly increased during the acute phase (p = 0.026) without any significant difference noted during convalescence (p = 0.271) as compared to controls. Both IL-3 and IL 4 levels were neither related to hemoglobin values, nor to red and white blood cell, neutrophil, lymphocyte and platelet counts. CONCLUSION: Serum IL-4 levels are increased in hospitalized children with symptomatic brucellosis, a finding suggestive of a Th2 response leading to severe infection. PMID- 11876514 TI - Intrasellar tuberculoma--a difficult diagnosis. AB - Tuberculosis is an exceptional cause of intrasellar mass lesion and diagnosis is usually established after histological examination following surgery. We report a 32-year-old woman with headache and amenorrhea, analytical features of hypopituitarism and an intrasellar mass lesion in radiological studies. A transsphenoidal approach was performed and tissue examination revealed pituitary tuberculoma. Additionally, we review the previously reported cases. PMID- 11876515 TI - Disseminated Nocardia asteroides presenting as pulmonary non-caseating granulomas in a patient with Waldenstrom macroglobulinemia. AB - Disseminated nocardiosis has never been described before in a patient with Waldenstrom macroglobulinemia. We report an unusual case of disseminated nocardiosis in a patient with Waldenstrom macroglobulinemia who presented with pulmonary non-caseating granulomas. The patient was successfully treated with trimethoprim-sulfamethoxazole (TMP-SMX) for 1 year. PMID- 11876516 TI - Failure of trimethoprim/sulfamethoxazole prophylaxis for Pneumocystis carinii pneumonia with concurrent leucovorin use. AB - Pneumocystis carinii is a common cause of pneumonia in patients with AIDS, however, the incidence has dropped with the availability of effective prophylactic regimens. First-line treatment for both acute Pneumocystis pneumonia and chronic prophylaxis is trimethoprim/sulfamethoxazole (TMP/SMX). This combination can cause hypersensitivity reactions as well as myelosuppression. The simultaneous administration of leucovorin during acute treatment has been shown to reduce the incidence of neutropenia, but may interfere with the efficacy of TMP/SMX. We report a case of P. carinii pneumonia in a patient with AIDS who failed TMP/SMX prophylaxis while taking leucovorin. PMID- 11876517 TI - Common symptoms--different diseases: coexistence of neurosyphilis and non Hodgkin's lymphoma. AB - We describe the case of a 32-year-old man with generalized lymphadenopathy who was diagnosed with a low-grade follicular small-cleaved cell lymphoma. The patient developed hearing loss, tinnitus and cerebrospinal fluid (CSF) pleocytosis attributed to central nervous system (CNS) infiltration by his malignancy, while receiving chemotherapy with vincristine, cyclophosphamide and prednisone. Despite intrathecal chemotherapy with methotrexate, the CSF pleocytosis persisted. Neurosyphilis was suspected because of prior history of gonorrhea and was confirmed with serologic studies of blood and CSF and from the decline of the anti-treponemal antibody titers with appropriate antibiotic therapy. Syphilis should be considered in the differential diagnosis of patients with generalized lymphadenopathy and neurologic signs or symptoms. PMID- 11876518 TI - Destructive osteoarthritis after delayed diagnosis of tuberculosis. AB - Osteoarticular tuberculosis rarely occurs in developed countries. Initial symptoms are often overlooked and the diagnosis is frequently delayed for several months. Thus, despite available diagnostic tools and accessible treatment, destruction of affected joints remains a complication of non-vertebral osteoarticular tuberculosis even in industrialized countries. We report a patient from Cleveland, Ohio, USA, in whom the delayed diagnosis of tuberculous osteoarthritis led to severe destruction of the left knee and finally, after superinfection with Staphylococcus aureus, to an above-the-knee amputation. The epidemiology, presentation, diagnosis and treatment of nonvertebral tuberculous osteoarthritis are discussed. PMID- 11876519 TI - Pyomyositis associated with bacillary angiomatosis in a patient with HIV infection. AB - Bacillary angiomatosis is an opportunistic infection occurring predominantly in patients with HIV infection. The manifestations of this vasculoproliferative disorder vary markedly. Virtually any organ site may be involved, singly or multiply. However cutaneous involvement can be a valuable clue to its diagnosis. We report a case of bacillary angiomatosis in an HIV-infected patient presenting as isolated pyomyositis of the right leg. The rarity of such a presentation and paucity of cutaneous lesions, as in our case, may render timely diagnosis elusive and difficult. Its recognition however, is important since bacillary angiomatosis, if untreated, is potentially fatal. PMID- 11876520 TI - Interaction between levothyroxine and indinavir in a patient with HIV infection. AB - Drug interactions are an important and emerging problem in the treatment of HIV infected patients. Protease inhibitors, like nonnucleoside reverse transcriptase inhibitors, are metabolized by the cytochrome P-450 enzyme system and each of these antiretroviral agents may interact with other drugs metabolized by this system. Some protease inhibitors may also interact with glucuronosyl transferase activity affecting plasma concentrations of drugs metabolized through this pathway. We describe a case of an HIV-infected patient, taking levothyroxine for hypothyroidism and clinically stable, who, after the introduction of an antiretroviral regimen containing indinavir, developed a pharmacological hyperthyroidism. PMID- 11876521 TI - Seroprevalence, viremia and genotype distribution of hepatitis C virus: a community-based population study in northern Italy. AB - BACKGROUND: Our aim was to define the burden of HCV infection and the related risk factors in a sample of the general population of northern Italy. MATERIALS AND METHODS: This is a survey on the prevalence of hepatitis C virus (HCV) infection carried out in a cohort (861 males and 1,293 females aged 17-67 years) from the general population of northern Italy. RESULTS: The crude HCV seroprevalence rate was 3.3%, higher in females than in males (p < 0.01) and with a clear age-related effect (test for linear trend: p < 0.01). When adjusted for sex and age in relation to the reference population, the prevalence was 3.7% (95% CI: 2.9-4.5%). In the univariate analysis, HCV infection was related to intravenous drug use (IVDU) (p < 0.001); transfusion (p < 0.001); tattooing (p = 0.02); chiropodist/manicure (p < 0.001); number of children (p < 0.01); educational level (p < 0.001). In the multivariate analysis sex, age over 50 years, transfusion, IVDU and tattooing were confirmed as risk factors, while the educational level seems to be a protective factor. Separate multivariate analysis for males and females points out that the age shift, the protective effect of education and the number of children as risk factor were present only in females. HCV viremia was detected in the 71.8% of the seropositive, resulting in an infection rate of 2.4% (95% CI: 1.7-3.0%). Genotype 1b accounted for 64.7% of isolates. CONCLUSION: In the general population of our area, HCV infection is mesoendemic and current infections are mostly sustained by genotype 1b. PMID- 11876522 TI - Loss of p14ARF expression in melanoma. AB - Lack of p14ARF expression or its functional inactivation has been observed in human and murine carcinomas. Although very few mutations of p14ARF have been detected in some cancer types, changes in expression seem to play an important role in the development of other human cancers such as mesotheliomas. To examine the p14ARF gene and expression of p14ARF protein in melanomas, we screened eight human melanoma cell lines and primary human melanocytes by RT-PCR, sequencing and immunoblotting. All melanoma cell lines analyzed expressed wild-type p14ARF mRNA as well as protein. P14ARF expression was investigated by immunohistochemical staining of 32 tissue samples of benign melanocytic nevi (n=14), melanomas (n=12) and melanoma metastases (n=6). In contrast to the results obtained from cell lines in vitro the immunohistochemical stainings revealed a correlation between the progression of melanoma and the lack of the p14ARF protein expression. Positive p14ARF protein staining was observed in 11 of 14 benign nevi, in 3 of 12 melanomas and in 0 of 6 melanoma metastases. In summary, we demonstrated a significant inverse correlation between p14ARF protein expression and progression of melanocytic tumors since the amount of p14ARF protein staining decreased from benign melanocytic nevi to metastatic melanoma in situ. These results suggest that p14ARF inactivation is important in the development of melanomas. PMID- 11876523 TI - The role of chemokines in allergic contact dermatitis. AB - Chemokines are important mediators of immune-mediated skin diseases. Allergic contact dermatitis (ACD) is the most thoroughly investigated T cell-mediated disorder because of the ability to easily reproduce the lesions in humans and the availability of an excellent mouse model. Migration of dendritic cells from the skin to lymph nodes is absolutely required for induction of hapten sensitization, and depends upon expression of CCR7 by mature dendritic cells and SLC in the lymph nodes. During expression of ACD, recruitment of T lymphocytes is driven by chemokines exposed on the surface of endothelial cells or released by activated resident skin cells such as mast cells, fibroblasts and keratinocytes. Chemokines are produced in a coordinated and sequential manner, with IL-8 and RANTES induced by TNF-alpha during early stages, and MCP-1, IP-10, Mig, I-TAC, I-309 and MDC induced by IFN-gamma during later stages. Infiltrating monocytes, dendritic cells and T cells are additional sources of chemokines for further leukocyte accumulation. Distinct T cell subsets express different chemokine receptors, with type 2 cells mostly attracted by eotaxin, MDC, TARC and I-309, and type 1 cells sensitive to IP-10, Mig, I-TAC, RANTES and MIP-1beta. MCP-1 is effective on both subsets. T regulatory cells, which inhibit dendritic cell function and are probably involved in the termination of ACD, are sensitive to MCP-1, MIPs and TARC, but express high levels of CCR8 and are more specifically attracted by I 309. Targeting chemokines and chemokine receptors may offer new opportunities for therapeutic interventions in ACD and other chronic inflammatory skin diseases. PMID- 11876524 TI - Identification of genes induced by rapid intraoperative tissue expansion in mouse skin. AB - A method of rapid skin stretching, i.e. hemispherical load cycling with an inflated subcutaneous silicone balloon (Rapid Intraoperative Tissue Expansion or RITE), permits the surgeon to rapidly elongate skin and create a flap of greater length for reconstructive plastic surgery. We have previously developed an experimental mouse model to evaluate RITE, and have shown that rapid stretching prevents ischemia and significantly reduces necrosis. Although the advantages of RITE have been demonstrated both clinically and experimentally, the cellular and molecular mechanisms underlying these benefits were unknown. In the study reported here, we used differential display reverse transcription polymerase chain reaction to identify genes that are specifically induced by RITE. Among four differential gene fragments, the expression of one was confirmed by Northern blot hybridization. The cDNA fragment was extended and the resultant sequence analyzed to reveal induction of truncated long interspersed nucleotide element 1 (LINE-1 or L1). Truncated L1 elements are located inside introns of many genes and among these genes myotubularin and insulin I are known to regulate cell growth. Northern hybridization using specific cDNA probes for myotubularin and insulin I demonstrated that it also was induced by RITE. This is the first reported study to show that L1, myotubularin and insulin I are responsive to rapid hemispherical and not rapid linear stretch. PMID- 11876525 TI - Liposome-encapsulated ursolic acid increases ceramides and collagen in human skin cells. AB - Skin wrinkling and xerosis associated with aging result from decreases in dermal collagen and stratum corneum ceramide content. This study demonstrated that ursolic acid incorporated into liposomes (URA liposomes) increases both the ceramide content of cultured normal human epidermal keratinocytes (NHEK), and the collagen content of cultured normal human dermal fibroblasts. In addition, URA liposomes increased the ceramide content of the skin of human subjects, with increases in hydroxy ceramides occurring after only 3 days of treatment. Both URA liposomes and retinoic acid decreased markers of keratinocyte differentiation (keratin 1, keratin 10 and involucrin) in cultured NHEK. Thus, URA liposomes have effects on keratinocyte differentiation and dermal fibroblast collagen synthesis similar to those of retinoids. However, this study showed that URA liposomes increase ceramides in NHEK, in contrast to the decreases previously shown to be caused by retinoids. URA liposomes have the potential to be used alone or in combination with other agents to restore or maintain skin ceramide and collagen content. PMID- 11876526 TI - Direct role of human dermal fibroblasts and indirect participation of epidermal keratinocytes in MMP-1 production after UV-B irradiation. AB - In vivo, matrix metalloproteinases are produced in response to ultraviolet B (UV B) irradiation and are considered to be involved in connective tissue alterations observed in photoaging. The respective roles of keratinocytes and fibroblasts in UV-B-induced MMP-1 production were investigated in monolayer cultures of keratinocytes and fibroblasts as well as in an epidermis model reconstructed in vitro. In contrast to fibroblasts, which secreted MMP-1 in response to UV-B irradiation, no accumulation of MMP-1 was observed after UV-B irradiation of keratinocytes. However, culture medium from UV-B-irradiated keratinocytes, which showed an increase in IL-1alpha and IL-6, induced MMP-1 production by human fibroblasts, suggesting that UV-B irradiation modulates MMP-1 production via both direct and indirect mechanisms. PMID- 11876527 TI - Protease nexin-1 messenger RNA levels are not affected by serum or interferon beta in cultured systemic sclerosis fibroblasts. AB - The aim of the present study was to analyze the effect of serum and human recombinant beta interferon (rIFNbeta) treatment on PN-1 mRNA levels in cultured dermal fibroblasts obtained from the skin of healthy donors and from lesional skin of systemic sclerosis (SSc) patients with the limited (CREST syndrome) or the diffuse form of SSc. Total RNA was isolated from fibroblasts derived from the skin of healthy individuals and from lesional skin of patients with CREST syndrome and the diffuse form of SSc cultured under different conditions (1% or 10% serum-supplemented medium) and treated with 500 IU/ml of rIFNbeta. PN-1 gene expression was assessed by Northern blot analysis. We detected variable PN-1 mRNA levels in normal control fibroblasts as well as in SSc fibroblasts under the different culture conditions (1% or 10% serum-supplemented medium). Accumulated PN-1 mRNA levels found in normal cultured fibroblasts were similar to or even higher than in SSc fibroblasts. PN-1 messenger levels were not significantly altered by IFNbeta treatment in normal or SSc cultured fibroblasts despite the presence of an IFN-stimulated responsive element (ISRE) in the promoter of the PN 1 gene. Our findings suggest that PN-1 expression in SSc fibroblasts at the mRNA level requires further investigation in a large number of SSc patients to better characterize the role of this serpin in the pathogenesis of SSc. We conclude that the transcriptional regulation of PN-1 is not associated with IFNbeta, an antifibrotic cytokine naturally produced by fibroblasts. PMID- 11876528 TI - The ornithine decarboxylase inhibitor, difluoromethylornithine, inhibits casein kinase II activity, c-Myc expression and normal human keratinocyte proliferation. PMID- 11876529 TI - Assessment of potential dermal and ocular toxicity and allergic properties of an extract of red propolis. PMID- 11876530 TI - The role of pioneer neurons in the development of mouse visual cortex and corpus callosum. AB - The primordial plexiform neuropil is very critical to neocortical development. The pioneer neurons, mainly Cajal-Retzius cells in the marginal zone, and subplate neurons in the subplate, differentiate from the primordial plexiform neuropil. In this study, the development of corpus callosum, visual cortex, and subcortical pathways has been observed in C57BL/6 mice with various methods, such as DiI labeling in vitro and in vivo, Dil and DiA in vitro double labeling, immunocytochemistry, and in vivo BrdU and Fast Blue labeling. As early as E14, the primordial plexiform neuropil can be found in the telencephalic wall, and it contains many pioneer neurons. On E15 the primordial plexiform neuropil differentiates into the marginal zone and the subplate. Cajal-Retzius cells exist in the marginal zone, and subplate neurons are in the subplate. Either Cajal Retzius cells or subplate neurons have long projections toward the ganglionic eminence, suggesting that they migrate tangentially from the ganglionic eminence. Cajal-Retzius cells are involved in radial migration, and subplate neurons participate to guide pathfinding of subcortical pathways. This study reveals how the pioneer neurons, through radial and tangential migration, play an important role in neocortical formation and in the pathfinding of the corpus callosum and subcortical pathways. Furthermore, DiI labeling in vivo has demonstrated the presence of pioneer neurons all along the corpus callosum pathway, especially in the midline. This suggests that pioneer neurons may also play a role in guiding the pathfinding of the corpus callosum. PMID- 11876531 TI - Modification of the rat aortic wall during ageing; possible relation with decrease of peptidergic innervation. AB - Structural changes of the male rat aorta were followed from birth to old age in male and female rats. In males, the vessel media width and area progressively increase concomitantly with a decrease of nuclei density during ageing, suggesting an hypertrophy of the smooth muscle cells. These correlations were however not evidenced in females. TUNEL-positive cells were found in media of 4 and 6 months in both sexes, mainly on the luminal side and in the adventitia. When biochemical markers were investigated with immunohistochemistry, media was uniformly stained by the anti-vimentin and anti-alpha-smooth actin at all stages investigated. On the contrary, the surface of media stained with anti-desmin decreased during ageing, especially on the luminal side. As observed with electron microscopy, with ageing the endothelium is replaced by small cells with pseudopodia adhering to the vestigial elastic lamina and infiltrating into the extracellular matrix left after the disappearance of smooth muscle cells. In addition, in the older rats (25-29 months) the elastic laminae are completely disorganised. Hypertrophy of the smooth muscle cells was confirmed by this approach. In parallel to this study, perivascular peptidergic innervation was stained with antibodies against calcitonin gene-related peptide (CGRP), substance P (SP), neuropeptide Y (NPY), and vasoactive intestinal polypeptide (VIP) at different ages during the whole life of rats. These peptides are present in stages younger than 6 months, then gradually disappear. In one year animals and older, the peptidergic innervation has totally disappeared. We discuss the possible role of peptidergic innervation in the control of the vessel wall cellular stability during ageing. PMID- 11876532 TI - Proliferation and differentiation of pituitary somatotrophs and mammotrophs during late fetal and postnatal periods. AB - Proliferation of somatotrophs and mammotrophs in the rat pituitary during late fetal and postnatal periods up to 4 weeks after birth was quantitatively studied with the double immunostaining of bromodeoxyuridine and the hormones produced by them. Somatotrophs were first detected in 18.5-day fetuses and rapidly increased in number throughout the periods studied. The cells labeled with both anti-BrdU and anti-GH were few in number until shortly before birth and then increased conspicuously during the first 10 days after birth. Mammotrophs were detected at gestational day 19.5 but they were few until the second week after birth, when their number began to increase rapidly. The percentage of the number of the cells double-labeled with both anti-BrdU and anti-GH to all somatotrophs was 8.3% at the most. This was about the same as that of corticotrophs during the late fetal period and that of thyrotrophs in the early postnatal period. In contrast, the percentage of double-labeled cells to all mammotrophs was 3.8% as a maximum, which is lower than the values for somatotrophs, corticotrophs, or thyrotrophs, indicating a smaller contribution of mitosis to mammotroph proliferation. It is possible that this smaller contribution is compensated for by transdifferentiation of cells committed to become the somatotroph lineage. However, coexistence of GH and PRL was not observed in the present material. PMID- 11876533 TI - Urokinase-type plasminogen activator (uPA) and matrix metalloproteinase-9 (MMP-9) expression and activity during early embryo development in the cow. AB - Several extracellular matrix (ECM)-degrading proteinases are hypothesised to play important roles during early mammalian development. In particular, urokinase-type plasminogen activator (uPA) and matrix metalloproteinase 9 (MMP-9) are expressed in peri-implantation mouse, sheep, and pig embryos and are implicated in the implantation process. These proteinases are not expressed in early (pre blastocyst) mouse, sheep or pig embryos. The aim of this study was to establish the gene expression and proteolytic activity of uPA and MMP-9 in in vitro produced (IVP) cow embryos. Using RT-PCR, mRNA transcripts for uPA and MMP-9 were detected during the first 7 days of development. To investigate the activity of these proteinases, conditioned media from various stages of development (days 2, 3, 4, 5 and 7) were assayed for uPA activity by chromogenic assay and MMP-9 activity by gelatin zymography. Both uPA and MMP-9 activities were detected in the media samples indicating the production and secretion of these proteinases. This pattern of proteinase expression is novel in comparison to the mouse where uPA and MMP-9 are only expressed from the blastocyst stage onwards. The results of this study suggest that these ECM proteinases have a role prior to implantation in the cow, in contrast to that exhibited by mouse, sheep and pig embryos. PMID- 11876534 TI - Expression of R-cadherin in the regenerating chick sciatic nerve. AB - The ultrastructural localization of R-cadherin in normal and regenerating chick sciatic nerves was investigated immunocytochemically, and was compared with that of N-cadherin. R-cadherin was found on the plasmalemmae of axons and Schwann cells where cell-cell contacts were made in the normal unmyelinated fibers. It was also noted that R-cadherin was expressed on the axolemmae where regenerating axons contacted with each other, and formed fasciculations. The normal myelinated fibers displayed no immunoreactivity except at the mesaxon. These findings of R cadherin were almost the same as those of N-cadherin of our previous study. N- and R-cadherin seemed to be co-expressed at the cell-cell contact points as mentioned above in the double labeling study. It is probable that cadherins contribute to the pathfinding of regenerating axons by causing them to form fasciculation. However, it seemed that there was no selective sorting of axons by N- and R-cadherin during regeneration as far as the present study was concerned. PMID- 11876535 TI - TUNEL staining and electron microscopy studies of apoptotic changes in the guinea pig vallate taste cells after unilateral glossopharyngeal denervation. AB - On the basis of our previous report that unilateral glossopharyngeal neurectomy in the guinea pig resulted in degeneration and disappearance of taste buds in ipsilateral vallate papillae (Huang and Lu 1996), it is reasonable to speculate that gustatory denervation may enhance apoptosis of taste bud cells, with taste buds decreasing in number and ultimately disappearing after neurectomy. We were therefore determined to investigate apoptosis of taste bud cells in guinea pig vallate papillae after unilateral glossopharyngeal neurectomy using both terminal deoxynuleotidyl transferase (TdT)-mediated dUTP nick-end labelling (TUNEL) at the light microscopic level and by conventional electron microscopy. A total of 34 adult guinea pigs were unilaterally glossopharyngeal-neurectomized and sacrificed at 3, 6, 12 h and 1, 3 and 7 days after surgery. The results revealed that only a very few TUNEL-positive nuclei indicating apoptosis were present in normal taste buds, but in surgically denervated papillae, they increased in number from 6 h-12 h after surgery, reached at peak on day 1 and then gradually decreased. In apoptotic cells from normal taste buds, electron microscopy revealed condensation of the chromatin against the nuclear envelope, changes in the nuclear envelope, and fragmentation of the nucleus, but the integrity of the plasma membrane and organelles was maintained. Neurectomized taste cells were also characterized by condensed and fragmentary nuclei, compactness of the cytoplasmic organelles, and the appearance of pedunculated protuberances on the cell surface. From these observations, we conclude that: (1) glossopharyngeal neurectomy enhanced apoptosis of vallate taste bud cells in guinea pig; (2) appropriate gustatory nerve innervation is an essential component for the maintenance of the taste bud, and may play a role in apoptosis of taste cells. PMID- 11876536 TI - Growth hormone (GH) action in early embryogenesis: expression of a GH-response gene in sites of GH production and action. AB - Growth hormone (GH) may act as a local growth factor in early embryonic development, since GH- and GH-receptor (GHR) immunoreactivity is present in all tissues and most cells of embryonic chicks during organogenesis. However, as GHR immunoreactivity could, alternatively, reflect the presence of GH-binding proteins (GHBPs) rather than authentic receptors linked to signal transduction mechanisms, GHR immunoreactivity may not be indicative of GH target sites. The possibility that GH may act as an autocrine or paracrine factor during embryogenesis was therefore assessed in the present study by determining the presence and cellular localization of mRNA for a GH-responsive gene. The mechanism of GH action involves the induction of a number of specific GH-response genes. In chickens a novel GH-responsive gene (GHRG-1) has been identified as a marker of GH action. In situ hybridization, using a 860 bp probe for GHRG-1 mRNA, demonstrated widespread expression of the GHRG-1 gene in embryonic tissues known to contain GH- and GHR-immunoreactivity (e.g. in the spinal cord, skin, heart, liver, muscle, bone and lung). GHRG-1 mRNA was not, however, present in all cells of each tissue. It was, furthermore, not present in subepithelial cells of the esophagus and bronchus and was lacking in many spinal cord ependyma, which are also known to lack GH immunoreactivity. These results therefore support the possibility that GH acts as an autocrine/paracrine factor during early chick embryogenesis, which was hitherto thought to be a "growth-without-GH" syndrome. PMID- 11876537 TI - Tumor angiogenesis as a prognostic factor in oral cavity carcinomas. AB - Currently, lymph node metastasis and thickness of the tumor are the gold standard as a predictor of survival in patients with oral cavity squamous cell carcinoma (OSCC). However, there is a significant correlation between microvessel density and the development of cervical metastases or recurrence. Previous studies have demonstrated that head and neck cancers are able to induce an angiogenic response in experimental models. This factor shows a strong correlation with regional recurrence. In this study we propose to use angiogenesis as an independent prognostic indicator of recurrence. We evaluated the expression of tumor angiogenesis in OSCC and determinated its possible usefulness as a prognostic factor. Thirty-three cases with diagnosis of OSCC were identified from January 1985 to January 1997 in the Head and Neck Department of the Instituto Nacional de Cancerologia in Mexico City. These cases were analyzed retrospectively for a minimum period of six months. All of them received a conventional complete treatment to the primary tumor and lymph node metastasis. Paraffin-embedded tumor specimens were available in all patients. The tumors were scanned and the areas of highest microvessel density (MVD) were immunostained for CD-34 using QBEnd/10 antibody. Statistical analysis included descriptive statistics, Wilcoxon test curves, and Cox's proportional hazards model for multivariate analysis. We identified 33 patients with OSCC, 16 were men and 17 women. The mean age among all patients was 58.9 years old. Based on tumor size 33.3% were T1, 27.3% T2, 12.1% T3, and 27.3% T4. The median microvessel count was 32.5. The mean percentage of MVD was 37 in patients with regional recurrence and in those patients without regional metastasis was 29 (p<0.05). 57.9% of the patients who presented recurrence had vessel counts over the median (p<0.01). In fact, 6 patients (46%) who showed more than 20% of angiogenesis expression and higher MVD presented with recurrence. Only 3 patients (23%) who had less than 20% of angiogenesis expression and lower MVD developed recurrence (p<0.01). Higher MVD was seen with increasing T and N stages; however, it did not show correlation with survival. In this study, angiogenesis expression demonstrated to be an independent factor of recurrence in patients with OSCC. It is suggested that it should be used as an independent prognostic indicator. In concordance with previous reports, we observed a significant correlation between MVD determination and recurrence of the tumor, followed by lymph node metastases and tumor size. PMID- 11876538 TI - Initial experience with sentinel lymph node mapping and biopsy in breast cancer. Preliminary results on 80 consecutive patients. AB - Sentinel node biopsy, in breast cancer, is a promising surgical technique for predicting histological findings in the remaining axillary lymph nodes, especially in patients with clinically node-negative breast cancer. 80 patients with breast cancer were evaluated for enrollment in this study. For mapping procedure 32 patients underwent lymphoscintigraphy in combination with an injection of vital blue dye, while 48 utilized only vital dye. In all patients, after sentinel node (SLN) biopsy, a complete axillary dissection was performed. The mapping procedure was possible in 79 cases (98.7%). The SLNs were positive in 27 (34.1%), in 52 patients SLNs were negative and in 50 of these all axillary nodes were negative. There was concordance in 77/79 cases (97.4%). The false negative rate was 3.8% (2/52). The overall sensitivity of the SNL biopsy was 93.1% (27/29), with a negative predictive value of 96.1% (50/52). This study demonstrates that accurate SLN identification was obtained combining lymphoscintigraphy and blue dye. Moreover, each method requires a suitable learning curve. Further studies are needed to define an accurate patient selection and the most speedy and precise method for intraoperative histological examination of SLNs. PMID- 11876539 TI - Detection of prostatic carcinoma: the role of TRUS, TRUS guided biopsy, digital rectal examination, PSA and PSA density. AB - The purpose of this study was to evaluate the efficacy of various diagnostic tests including transrectal ultrasound (TRUS), TRUS guided biopsy, digital rectal examination (DRE), prostate specific antigen (PSA), and prostate specific antigen density (PSAD) in detecting prostatic carcinomas. One hundred and thirty-four men underwent TRUS guided random, or directed and random sonographic biopsies of the prostate. The mean age was 64.67 (range, 31- 88) years. Indications for biopsy were abnormal findings suggesting prostatic carcinoma on DRE or increased levels of PSA, defined as 4.0 ng/ml or greater in a monoclonal antibody assay. PSAD was calculated by dividing the serum PSA in ng/ml to the volume of the entire prostate in cm3. The biopsy results were grouped as benign, malign and, prostatitis. The patients were also divided into three groups according to their PSA values. Of the 134 patients evaluated, 31 (23.1%) had prostate adenocarcinoma, 89 (66.4%) had benign prostatic tissue, hyperplasia or prostatic intraepithelial neoplasia, and 14 (10.4%) had prostatitis. The mean PSA and PSAD of the carcinoma group were significantly higher than those of the noncancer group. In the group of patients with PSA levels between 4 and 10 ng/ml, abnormal TRUS or DRE increased cancer detection rate, where neither PSA nor PSAD was capable of discriminating the patients with and without cancer. PSAD did not prove to be superior to the other diagnostic tests in this study. We recommend biopsy when either TRUS or DRE is abnormal in patients with PSA levels between 4 and 10 ng/ml. In the patients with PSA levels greater than 10 ng/ml, biopsy is indicated whatever the findings on TRUS or DRE are, since cancer detection rate is high. PMID- 11876541 TI - Surgical treatment of recurrences of pleomorphic adenoma of the parotid gland. AB - From 1989 to 1999 a retrospective study was conducted on 7 patients with recurrent pleomorphic adenomas of the parotid gland who were referred to the MaxilloFacial Department of the University of Rome "La Sapienza" after having undergone surgery elsewhere. The mean time interval between the first operation and recurrences ranged from 15 months to 13 years, and the average time interval was 7.7 years. Implantability of the lesion and inadequate surgery that produced rupture of tumour capsule and tumour cells bleeding into surrounding glandular parenchyma, were the reasons for tumour recurrence. The instrumental examinations used for planning the surgical treatment to be applied and for studying the relations of recurrence with glandular parenchyma were CT (with contrast medium) or MR of head and neck. These patients underwent total parotidectomy with facial nerve preservation and no recurrence occurred in any patient. The results of this study underscore the importance of adequate surgical excision of initial recurrences as well as primary tumours to prevent tumour recidivism. Finally, tumour control rates and facial nerve preservation are enhanced with formal parotidectomy for recurrent tumour when feasible. PMID- 11876540 TI - Correlations between nuclear/cytoplasmic area ratio and classification of cervical smears. AB - The aim of this study was to assess the nuclear/cytoplasmic (N/C) ratio using computerized image analysis of cervical smears with intraepithelial neoplasia (CIN) grade I to III associated or not with cellular changes of human papillomavirus (HPV) in an attempt to determine if this method is more sensitive for the estimation of the grade of CIN. One hundred and ten cervical smears from women with a mean age 35.03 years were studied. The cytological diagnosis was as follows: CIN I + HPV (11), CIN II + HPV (11), CIN II + HPV (8), CIN I (7), CIN II (6), CIN III (8), Ca (22), HPV (32), CIN I-II + HPV (2) and CIN II-III + HPV (3). All cases were histologically examined: 93 cases were in agreement and 17 were under- or overestimated cytologically. The morphometric study of cervical smears was carried out by image analysis. Data were analysed by one way analysis of variance followed by Bonferroni test of multiple comparisons. Statistically significant differences were detected between the three grades of CIN or CIN HPV or only HPV (p<0.0001). The results demonstrated that the N/C ratio measured by image analysis on precancerous lesions of cervical smears could be considered as an additional tool for the classification of cervical smears, especially in determining the discrepancies between cytological and histological diagnoses. PMID- 11876542 TI - Non-small cell lung cancer. The role of chemoimmunoradiotherapy after surgery. AB - The aim of this study was to determine expediency of adjuvant chemoimmunoradiotherapy for radically operated non-small cell lung cancer patients (LCP) with pathologic stage II-III (T1-4N0-2M0G1-3). In retrospective trial (1985-1998) a 5-year survival of 54 consecutive radically operated LCP after adjuvant chemoimmunoradiotherapy (group A) was compared with 5-year survival of 264 LCP, after radical procedures (group C) and with 5-year survival of 86 radically operated LCP after postoperative radiotherapy (group B) (45-50 Gy). I cycle of chemoimmunotherapy was given on day 10-14 after complete resections. Radiotherapy (45-50 Gy) was administered since day 7 after 1 cycle. After irradiation 3-4 courses of CAVT were repeated every 21-28 day. Variables selected for 5-year survival and life span study were sex, age, TNMG, cell type, tumor size. Survival curves were estimated by the Kaplan-Meier method. Differences in curves between groups of LCP were evaluated using a log-rank test. Multivariate proportional hazard Cox regression, multi-factor clustering, structural equation modeling and Monte Carlo simulation were used to determine any significant overall regularity. 5-year survival was superior in group A (64.8%: 35 out of 54 LCP with N0-2; life span=1998.2+/-156.9 days) compared with group B (45.3%: 39 out of 86 LCP with N0-2; life span=1296.4+/-109.5 days) (P<0.001). 5-year survival of group C was 63.6% (168 out of 264 LCP with N0-2; life span=1738.3+/-63.4 days) (P>0.05 for group A and P<0.001 for group B). For LCP with N1-2 5-year survival was significantly superior for group A (63.6%: 21 from 33; life span=1934.0+/-180.9 days) compared with group C (28.1%: 25 out of 89; life span=1056.9+/-91.1 days) (P<0.001) and with group B (35.6%: 21 out of 59; life span=1051.7+/-119.6 days) (P<0.001). Structural equation modeling and Monte Carlo simulation confirmed significant overall differences between 5-year survival (P<0.05) and life span (P<0.001) of LCP with N1-2 in group A with respect to group C or B; however, 5-year survival of LCP for N0 in groups A, B and C were not significantly different. PMID- 11876543 TI - Bone marrow toxicity in mice treated with indium-114m-labelled blood cells. AB - Clinical trials with autologous indium-114m-labelled lymphocytes have revealed significant anti-tumour effects in chronic lymphocytic leukaemia patients with highly resistant disease. Substitution of the lymphocyte vector with heat-damaged red blood cells (HDRBC) may make this treatment more universally applicable and reduce the dose-limiting myelosuppression encountered with labelled lymphocytes. Therefore, the bone marrow localization and toxicities of indium-labelled lymphocytes or HDRBC have been investigated in BDFI mice. At 24 hours approximately 4% and 1.2% of 114In(m) administered as labelled lymphocytes or HDRBC respectively was localized within the bone marrow and remained constant for 57 days thereafter. Toxicity towards bone marrow stem cells, measured as CFU-S, was equivalent for both cellular vectors. However, at clinically relevant activities, 114In(m) HDRBC were less toxic than labelled lymphocytes towards committed progenitors, assayed as in vitro-CFC and CFU-Meg. These data suggest that substitution of HDRBC for lymphocytes as the 114In(m) vector may be beneficial in reducing the myelosuppression associated with this technique. PMID- 11876544 TI - Boron compounds against human leukemic cells. AB - Two new boron compoumds, dihydroxy(oxybiguanido) boron (iii) hydrochloride monohydrate (HB) and guanidine biboric acid adduct (GB) were used in this study to observe the antitumor effect. Leukemic blast cells isolated from chronic myeloid leukemia (CML) patients showed significant cell growth inhibition within twentyfour hours. IC50 of GB and HB was 2mg/ml. The metabolically active cells were found to be inhibited by drug treatment as assessed by MTT test. Inhibition of 3H Thymidine incorporation also supported the above result. In this study we investigated the molecular mechanisms by which HB and GB induce apoptosis in immature blast cells. PMID- 11876545 TI - The different level of immunological recovery after chemotherapy in leukemia and lymphoma patients. AB - Analysis of T lymphocyte subsets made a great progress in attempting their important role in host defence against malignant disease. As the major regulatory elements in the immune system, helper (CD4+) and suppressor (CD8+) T cells are required for recognizing a large scale of surface antigen on tumor cells and subsequently, for mediating the regression of cancers. Many data suggest that T cell immunity system is not intact in different malignant diseases. The malignant disorders of hemopoietic tissue are frequently associated with T cell functional impairment, the mechanism(s) of which is still unresolved. In recent years, defects in immunoregulatory T cell network have been repeatedly proposed as the cause of this alteration and might be considered as a parameter of immunodeficiency in those patients. The additional disturbance of T cell immunoregulatory system may be induced by different extrinsic factors. In this context the chemotherapeutic agents with their non-selective action, including an immunomodulatory activity, should be involved in the evaluation of lymphocyte subset changes during treatment. In this brief review we analyzed the data concerning the T cell subset changes, both numerical and functional, before and in course of the treatment of patients with different hematologic malignancies. According to this consideration, it might be suggested that the immune system of these patients have some capacity to recover from the suppressive effect of chemotherapy, although not sufficiently to be normalized. The long-term monitoring of a larger scale of immunological parameters may probably help us to resolve whether T lymphocyte subset changes are related only to the therapy or, in some instance, might be connected with compromised natural host defence mechanisms in oncohematologic patients. PMID- 11876546 TI - Soluble immunological parameters and early prognosis of renal cell cancer patients. AB - In local or metastatic cancer, a prognostic tumour marker could be a valuable tool in the selection of different treatments. In renal cell cancer (RCC) no such markers have been available. We therefore evaluated the association between several pretreatment serum markers, tumour classification and short term survival in RCC patients. Serum samples were collected before surgery and three months thereafter from 24 RCC patients. Interleukin-6 (IL-6), IL- 12, soluble IL-2 receptor (sIL-2R) and intercellular adhesion molecule-1 (sICAM-1) were measured in serum samples using specific commercial enzyme immunoassay kits. Serum IL-6, sIL-2R and sICAM-1 levels before nephrectomy were significantly higher in non local tumours than in local ones (mean IL-6 53 pg/ml versus 6.3 pg/ml, and sICAM 1 443 ng/ml versus 290 ng/ml, sIL-2R 3779 pg/ml versus 1796 pg/ml). In contrast, IL-12 levels were higher in local tumours (148 versus 102 pg/ml) and the levels increased significantly (P < 0.005) after removal of the primary tumour in patients with local disease. All patients with local tumours had normal IL-6 values, while only one with a non-local tumour had IL-6 levels below 10 pg/ml. In addition, IL-6 and sICAM-1 levels before operation were significantly higher in patients with short (less than one year) survival (p=0.007 to IL-6 and p=0.006 to sICAM-1). In contrast, patients with shorter survival had significantly lower IL 12 (p=0.03) levels. Our findings suggest that RCC induces changes in several immunological parameters. These soluble immunological factors, IL-6, IL-12, sIL 2R and sICAM-1, might have a role as prognostic factors in RCC. PMID- 11876547 TI - Detection of tyrosinase mRNA by an optimised nested RT-PCR in the peripheral blood of patients with advanced malignant melanoma. AB - Malignant melanoma (MM) has a high metastatic potential even in small primary lesions, and an early distinction between localized and regionally/distally advanced disease is of major importance for the patients' treatment and, consequently, for their survival. Exploiting the fact that tyrosinase is a tissue specific enzyme which is only expressed in normal skin melanocytes and MM cells that invade the blood during metastasizing, the objective of our study was to optimise the nested RT-PCR assay for the detection of tyrosinase mRNA and, hence, to detect circulating melanoma cells (CMC) in whole venous blood of MM patients. Eighteen MM patients (stage III and IV, according to AJCC) and 8 healthy subjects were included in our study. Following optimisation of the procedure, the lowest detection limit of 10 MM cells per 1 ml of the blood was achieved. Tyrosinase mRNA was detected in 27.8% (5/18) of blood samples from MM patients and in none of the healthy volunteers. Preliminary results of this study suggest that the method is sensitive and specific to the CMC detection in the peripheral blood and may thus be helpful in determining the disease stage and, consequently, in planning treatment. PMID- 11876548 TI - Nuclear/Nucleolar morphometry and DNA image cytometry as a combined diagnostic tool in pathology of prostatic carcinoma. AB - Paraffin tissue sections from 50 patients with prostate adenocarcinoma were used to study nuclear and nucleolar morphometric features by image analysis. The results were compared to DNA ploidy and Gleason grade. In the examined histological samples nuclear and nucleolar areas were positively interrelated. It was also noticed that the higher the percentage of nucleolated nuclei, the bigger the nuclear and nucleolar areas. The morphometric characteristics did not differ significantly among the four grades of the examined specimens. In well differentiated carcinomas the DNA index was lower than in the rest at a statistically significant level. Hypodiploid carcinomas were found to possess significantly bigger nuclear areas than any other DNA index group. Morphonuclear evidence of anaplasia and DNA aneuploidy may be used as diagnostic tools in prostate cancer in addition to Gleason grade. PMID- 11876549 TI - Prospective study of Ki-67 labeling index in the mucosa adjacent to cancer as a marker for colorectal cancer metastasis. AB - Multivariate regression analysis has shown that Ki-67 labeling index in the mucosa adjacent to cancer was the most significant marker for colorectal cancer metastasis among several metastasis-related parameters according to our previous retrospective data base (7). We have performed a prospective study to ascertain whether Ki-67 labeling index in the mucosa adjacent to cancer is a useful preoperative diagnostic marker for colorectal cancer metastasis. In 182 registered cases colonoscopically biopsied, we performed surgical resection of the cancer in 37 adenocarcinoma cases, which were registered in the study. In 31 out of the 37 cases except for 6 cases with an insufficient amount of non neoplastic mucosa, preoperative diagnosis for metastasis was performed using the Ki-67 cutoff line. The cutoff line was set at 15% according to our previous retrospective database. The preoperative diagnosis for metastasis was compared to the pathological findings of the resected specimen. The incidences of correct diagnosis for metastasis was 81% (25/31). There were 3 false positive cases and 3 false negative cases in Dukes' A-B and Dukes' C, respectively. The mean Ki-67 labeling index in the mucosa adjacent to cancer of Dukes' A-B and Dukes' C-D cases, except for the 6 misdiagnosed cases, was 7.4+/-5.0% and 29.9+9.8%, being significant at p<0.0001 by unpaired Mann-Whitney U test. These results suggest that Ki-67 labeling index in the mucosa adjacent to cancer might be a good marker for metastasis in colorectal cancer. PMID- 11876550 TI - Cdk1/P34Cdc2 and P21waf expression in colorectal adenomas and carcinomas. AB - p21WAF1,CIP1,SDI 1 is a specific inhibitor of cyclin-dependent kinase and a tumor suppressor involved in the pathogenesis of a variety of malignancies. Cdk1/p34cdc2 plays a crucial role during mitosis. The immunoreactivity of p21 WAF1,CIP1,SDI 1 and cdk1/p34cdc2 was evaluated in adenomas and adenocarcinomas of the human large bowel. The expression of p21WAF1,CW1,SDI 1 was detected in 5 out of 15 adenomas (33.3 %) and in 13 out of 36 adenocarcinomas (36 %). Cdk1/p34cdc2 was expressed in 11 out of 15 adenomas (73.3%) and in 18 out of 36 adenocarcinomas (50%). In conclusion, our results indicate that there is an expression of cdk1/p34cdc2 in colorectal adenomas and absence of p21 WAF1,CIP1,SDI 1 in both colorectal adenomas and carcinomas. These results suggest that the proliferative activity of colonic cancer might not be solely dependent on control of the cell cycle by p21 WAF1,CIP1,SDI Cdk1/p34cdc2 expression, in a high percentage of both adenomas and adenocarcinomas suggest that this phenomenon may be an early event in colorectal carcinogenesis. PMID- 11876551 TI - Frequent alterations of the beta-catenin and TCF-4 genes, but not of the APC gene, in colon cancers with high-frequency microsatellite instability. AB - High-frequency microsatellite instability (MSI-H) due to defective DNA mismatch repair (MMR) is a characteristic of the majority of tumors from kindreds with hereditary nonpolyposis colorectal cancer (HNPCC) and a subset of sporadic cancers. To better understand the molecular characteristics of colon cancers with MSI-H, we analyzed these cancers for alterations of genes, such as APC, beta catenin, and TCF-4 genes, involved in the Wnt signaling pathway. Following the National Cancer Institute (NCI) criteria, 385 unselected colon cancers were classified as follows: 50 (13%) MSI-H tumors, 36 (9%) low-frequency MSI (MSI-L) tumors, and 299 (78%) microsatellite stable (MSS) tumors. The frequency of APC mutations was significantly lower in MSI-H tumors (9 out of 50) than in MSI-L (12 out of 20) and MSS (66 out of 100) tumors (P = 0.0005 and P < 0.0001, respectively). In contrast, the frequency of exon 3 mutations in the beta-catenin gene was higher in MSI-H tumors (10 out of 50) than in MSI-L tumors (0 out of 30; P = 0.0110) and MSS tumors (3 out of 100; P = 0.0010). Frameshift mutations in a (A)9 tract of the TCF-4 gene were detected in 44% (22 out of 50) of MSI-H tumors, but not in any of the 20 MSI-L tumors or 40 MSS tumors. In total, 78% of MSI-H tumors and 84% of the remaining tumors had at least one alteration in APC, beta catenin, or the TCF-4 genes. Although further analysis is needed to functionally characterize the consequences of each of these alterations on beta-catenin/TCF target gene expression, our results suggest that the activation of the Wnt signaling pathway plays a pivotal role in colon tumorigenesis, irrespective of MSI status. PMID- 11876553 TI - A C/T polymorphism in the urokinase-type plasminogen activator gene in colorectal cancer. AB - Urokinase plasminogen activation system can play an important role in the appearance and progression of many cancers. Urokinase-type plasminogen activator (uPA) is implicated in the control of cell adhesion and invasion, and is regarded as a strong prognostic marker in colorectal cancer. A C-->T substitution (the C/T polymorphism) in the nucleotide sequence encoding the kringle structure of uPA results in an alteration from proline to leucine at position 121. This substitution may be directly or indirectly involved in the decreased affinity for uPA substrates. In the present work the distribution of genotypes and frequencies of alleles of the C/T polymorphism were investigated. Tumour tissues and distal mucosa samples were obtained from 40 patients with colorectal cancer. Blood samples from sex and age matched healthy individuals served as control. The C/T polymorphism was determined by PCR amplification using the allele specific primers. No differences between genotypes of the C/T polymorphism in cancer tissue and distant mucosa of each patient were found. The distributions of the genotypes in both patients and control differed significantly (p < 0.05) from that predicted by the Hardy-Weinberg distribution. A distinct preference of heterozygotes (70% - patients, 65% - controls) was observed in both patients and controls. Additionally, there were no differences in the frequencies of the C and T alleles in both groups. The C/T polymorphism of the uPA gene may not be linked with colorectal cancer. PMID- 11876552 TI - Establishment and characterization of a human rectal neuroendocrine cell carcinoma in vitro. AB - Human colorectal neuroendocrine cell carcinoma ( NECC ) is uncommon. Treatment of the disease has not yet been established, and NECC of the colon and rectum behave clinically more aggressively than their exocrine counterparts, so the prognosis is generally worse. One reason for the lack of established treatment is that there are no model systems of this disease. There have been a few reports on cell lines from neuroendocrine tumors, because these tumors are difficult to culture, and there are even fewer reports on colorectal carcinoma cell lines with neuroendocrine features. We therefore attempted to establish a permanent cell line in order to investigate the biological behavior and treatment of NECC. The cell line we succeeded in culturing is called N-TAK1. Gastrin promotes the growth of gastrointestinal epithelial cells and also stimulates the growth of gastrointestinal cancers. Hormone-receptor antagonists restrict the growth of hormone-dependent tumors. The growth of colon cancer was promoted by the application of gastrin, whereas it was restricted by proglumide, which is known to be a gastrin receptor antagonist. We demonstrated that gastrin has a stimulatory effect on the growth of N-TAK1 cells and that it could be detected by immunohistochemistry in the cells. We also showed that proglumide inhibited the growth effect of gastrin. PMID- 11876554 TI - Hypothalamic digoxin mediated model for oncogenesis. AB - This study assessed the changes in the isoprenoid pathway and its metabolites digoxin, dolichol and ubiquinone in neoplasms (CNS astrocytomas - glioblastoma multiforme and high grade non - Hodgkin's lymphoma). The following parameters were assessed-isoprenoid pathway metabolites, tyrosine and tryptophan catabolites, glycoconjugate metabolism, RBC membrane composition and free radical metabolism. There was an elevation in plasma HMG CoA reductase activity, serum digoxin and dolichol and a reduction in RBC membrane Na+-K+ ATPase activity, serum ubiquinone and magnesium levels. Serum tryptophan, serotonin, nicotine and quinolinic acid were elevated while tyrosine, dopamine, noradrenaline and morphine were decreased. The total serum glycosaminoglycans and glycosaminoglycan fractions (except dermatan sulphate in the case of CNS astrocytomas), the activity of GAG degrading enzymes and glycohydrolases, carbohydrate residues of glycoproteins and serum glycolipids were elevated. HDL cholesterol showed a significant decrease and free fatty acids & triglycerides were increased. The RBC membrane glycosaminoglycans, hexose and fucose residues of glycoproteins and phospholipids were reduced. The activity of all free radical scavenging enzymes, concentration of glutathione, iron binding capacity and ceruloplasmin decreased significantly while the concentration of malondialdehyde (MDA), hydroperoxides, conjugated dienes and NO increased. The concentration of alpha tocopherol was unaltered. Membrane Na+-K+ ATPase inhibition due to elevated digoxin, altered membrane structure and digoxin related tyrosine / tryptophan transport defect leading to increased levels of depolarising tryptophan catabolites and decreased levels of hyperpolarising tyrosine catabolites can lead to alteration in intracellular calcium/magnesium ratios and oncogene activation. Intracellular magnesium deficiency can produce defective microtubule related spindle fibre dysfunction and chromosomal non-dysjunction contributing to neoplastic cellular polyploidy and aneuploidy. Digoxin induced tryptophan/tyrosine transport defect can alter neurotransmitter patterns with increased serotonin, quinolinic acid, nicotine & glutamatergic transmission and reduced dopamine, morphine and noradrenaline levels leading to oncogenesis. Glycoconjugate metabolism is altered by elevated dolichol levels and magnesium depletion consequent to Na+-K+ ATPase inhibition. There is a qualitative alteration in proteoglycans and glycoproteins, defective membrane formation and structure and reduced lysosomal stability leading to disordered contact inhibition and tumour antigen presentation contributing to oncogenesis. Digoxin induced alteration in intracellular calcium/magnesium ratios and low ubiquinone levels can lead to a mitochondrial dysfunction resulting in increased free radical generation and reduced scavenging & caspase-3 activation producing a P21 defect contributing to oncogenesis. PMID- 11876555 TI - Role of alphavbeta3 integrin receptors in breast tumor. AB - The intriguing problem of tumor metastasis involves adhesion of the metastatic cells via the integrin group of cell surface receptors to the ECM proteins of the basement membrane. Modulation of integrin family of cell surface receptors is believed to be associated with stages of tumor metastasis. In this present study, results obtained indicate that alphavbeta3 vitronectin receptors may play an important role in the malignant behaviour of human breast tumor. Alphavbeta3 integrin receptors can modulate the activity of collagenase enzyme and thereby the invasive property of breast tumor cells. PMID- 11876556 TI - Nitric oxide-mediated antitumor activity induced by the extract from Grifola frondosa (Maitake mushroom) in a macrophage cell line, RAW264.7. AB - We have investigated D-fraction (MDF) extracted from Grifola frondosa (Maitake mushroom) on the inducible nitric oxide synthase (iNOS)-mediated nitric oxide (NO) production in RAW264.7 (RAW) cells, a murine monocyte/macrophage cell line, with special reference to antitumor activity of MDF against human hepatoma derived huH-1 cells. MDF could induce iNOS mRNA expression in RAW cells in a dose range of more than 30 microg/ml, but the effect of 10 microg/ml of MDF was negligible. The iNOS mRNA expression induced by 100 microg/ml of MDF was 6 hrs later, but lasted for a longer time than that of lipopolysaccharide (LPS), a representative iNOS inducer. Although iNOS mRNA levels in MDF-stimulated cells were almost equal to LPS-stimulated cells at the peak time, the cumulative amount of nitrite was only about 50% compared with that of LPS-treated cells. When huH-I cells were cultured in MDF containing media in a 24-well plate with inserted porous bottom in the presence or absence of RAW cells, the viability of huH-1 cells decreased significantly only in the presence of RAW cells in MDF dose dependent manner. This antitumor activity of RAW cells in the presence of MDF was abolished or attenuated by the addition of L-NAME, a NOS inhibitor, confirming that this phenomenon is due to iNOS-mediated NO production by RAW cells, but not direct cytotoxic activity of MDF against huH-1 cells. These data suggest that MDF is a novel inducer for iNOS which contributes at least in part to antitumor activity of MDF. PMID- 11876558 TI - A surgical approach on an emergency basis for removal of Wilms' tumor with intracardiac neoplastic extension. AB - Extension of Wilms' tumor through inferior vena cava into the heart two years after the completion of therapy, is a very rare clinical aspect. We successfully operated on a 7 year old girl who had a huge right atrial mass which filled the right ventricle and right ventricular out flow tract, on an emergency basis using cardiopulmonary bypass. PMID- 11876557 TI - Effect of purified glutaminase from human ascites fluid on experimental tumor bearing mice. AB - Glutamine is the major respiratory fuel and energy source of the rapidly proliferating tumor cells and that is why glutamine clearance by glutaminase therapy provides an opportunity to fight against the neoplasm. Glutaminase from bacterial source was tried on experimental models but had to be excluded because of its limited efficacy. Search for a better glutaminase continued exploiting the mammalian sources. In the present study, glutaminase purified from human ovarian cancer ascites fluid was used in experimental solid and ascites mice model alone and in combination with Cu-Sulphate and heparin. Cumulative findings indicate that the enzyme alone is quite effective in lowering tumor burden and reducing not only the tumor induced angiogenesis, but also an angiogenic inducer, heparin mediated angiogenesis. However, the presence of Cu with the enzyme, amplified the antineoplastic response by improving anti-angiogenic potential and hematological status of the tumor bearing host. Therefore, Cu-glutaminase combination strengthened the hypothesis that together they may provide a better therapeutic regimen in experimental mice tumor model. PMID- 11876559 TI - Hemangioma of the breast: an unusual lesion without univocal diagnostic findings. AB - Hemangiomas of the breast, either capillary or cavernous, are thin-walled, blood filled vascular spaces, separated by fibrous septa, with extensive fibrosis and sometimes phleboliths. Clinical diagnosis is rather difficult. Generally they are coincidental microscopic findings. We report herein a case of breast hemangioma misdiagnosed at ultrasound and mammography. A 63-year-old woman described intermittent sharp pain in the right breast. Physical examination, mammography and ultrasonography were not sufficient for the diagnosis. Surgical excision of the lesion was performed. At histology it was found to be a cavernous hemangioma without cellular atypia. The patient is now 9 months post surgery and is well. The role of the single diagnostic examination is limited. The Authors believe the complementary role of all available techniques in the evaluation of a breast lesion. PMID- 11876560 TI - ALK-negative, T-cell anaplastic large cell lymphoma with hypocellular and neutrophil-rich features. PMID- 11876561 TI - Vital statistics as a data source. AB - A focus group convened at a National Institutes of Health (NIH) Stillbirth Conference discussed issues related to vital statistics as a data source on fetal mortality. Fetal death is a tragic occurence and part of the continuum of pregnancy outcomes. A primary source of data is the National Vital Statistics System fetal death component, which is routinely used to track mortality trends and to provide the context for nonrepresentative investigations. The data are also used to examine differentials in mortality by characteristics and to evaluate data quality. Issues concerning how to improve the vital statistics data include culture and context; data instrument; persons providing information; what information is available to the information provider; processing issues; and use of data. Perhaps most important is to enhance recognition of fetal death both for the individual confronting a loss and to promote research. Other studies are needed to augment the information obtainable through vital records. PMID- 11876562 TI - Contribution of late fetal deaths to US perinatal mortality rates, 1995-1998. AB - We analyzed US fetal death and linked infant birth-death certificate data for 1995-1998 to evaluate perinatal deaths (late fetal deaths [> or = 28 weeks' gestation] and early neonatal deaths [< or = 7 days of life]) by race, Hispanic ethnicity, state of residence, and selected demographic characteristics. We also compared components of perinatal mortality, late fetal deaths, and early neonatal deaths, by birthweight, gestational age, and selected maternal medical conditions during pregnancy. From 1995 through 1998, there were 221,767 fetal deaths at > or = 20 weeks' gestation and infant deaths at less than 1 year. Of these, 113,421 (51%) were perinatal deaths; late fetal deaths accounted for 47% of perinatal deaths. The total perinatal mortality rate declined 5.3%, from 7.5 to 7.1 per 1,000 live births plus late fetal deaths. Blacks experienced higher perinatal mortality rates than whites (rate ratio = 2.1). Among perinatal deaths > or = 28 weeks' gestation, the ratio of fetal to neonatal deaths was 3.4 among blacks and 2.4 among whites. State-specific rates ranged from 5.2 to 13.1 per 1,000 live births plus late fetal deaths. Although late fetal deaths are not included in routine statistics of pregnancy outcomes, these deaths represent a large proportion of adverse pregnancy outcomes. Surveillance of perinatal mortality provides a more complete picture of the health of women, fetuses, and newborns. Improving the quality of surveillance data regarding fetal deaths is essential for more effective use of these data. This information can be used to prevent excess perinatal deaths and reduce disparities in pregnancy outcomes among high risk subgroups identified by individual and population characteristics. PMID- 11876563 TI - The epidemiology of stillbirth. AB - Stillbirths account for an increasing proportion of feto-infant mortality. Yet, causes of stillbirth are rarely reported, and the causes of most stillbirths remain unknown. Few studies focus specifically on the epidemiology of stillbirth. Major risk factors include high maternal age, smoking, and overweight. The prevalence of delayed childbearing and, especially overweight, are increasing in most developed countries. The proportion of stillbirth attributable to overweight is likely to increase. PMID- 11876564 TI - The national fetal death file. AB - The most comprehensive source of US data on fetal deaths of 20 gestational weeks or greater is available through the National Vital Statistics System (NVSS). The NVSS is a collaborative effort between the independent reporting areas (the individual States and the territories), and the federal government or its agent, the Centers for Disease Control, and Prevention's National Center for Health Statistics (NCHS). The federal government has no authority to register vital events. The registration of births, deaths, fetal deaths, marriages, and divorces is solely a state responsibility. However, NCHS is mandated by law to produce national data based on vital events. To promote the uniformity necessary to create a national file from this decentralized system, NCHS attempts to influence state systems via the development of certain standards, primarily, The Model State Vital Statistics Act and Regulations (The Model Law), and the Standard Certificates and Reports. The Model Law definitions for live birth, fetal death and induced termination of pregnancy are based on international standards set by The World Health Organization. All states have definitions of fetal death consistent with the Model Law. The Model Law also recommends reporting requirements for fetal death, but state requirements vary. This variation results in differences in reporting of fetal deaths among areas. Other limitations to the national fetal death file include: the under-reporting of fetal deaths incidence, higher than acceptable levels of missing data for some items, and the accuracy of the data reported. Also of concern is the potential misclassification of fetal deaths and short-lived live births. These limitations are amenable to improvement. The upcoming revision of the US Standard Report of Fetal Death addresses these issues and offers an opportunity to strengthen the quality of fetal death data. The development of worksheets, detailed specifications and instruction manuals, and a reformatted cause of death section should importantly enhance the quality of national fetal death file and ultimately reduce the incidence of these tragic events. PMID- 11876565 TI - Methodologic considerations for population-based research on fetal deaths: overcoming data gaps. AB - Fetal deaths comprise a large component of perinatal mortality and remain an understudied pregnancy outcome, especially from a population perspective. Interpretation of the findings from clinical or community-based studies can be difficult without a clear understanding of fetal death at the population level. This article addresses the critical data gaps underlying population-based research on fetal deaths, including the magnitude and scope of the problem, the probability of occurrence, the populations at risk, and the importance of accounting for prior reproductive history. Suggestions are given for new avenues of population-based research such as prospective inquiry of couples attempting pregnancy and newer analytic and modeling strategies for assessing risk and for addressing the lack of independence in pregnancy outcomes. PMID- 11876567 TI - Maternal medical disease: risk of antepartum fetal death. AB - Although certain maternal medical conditions increase the risk of antepartum fetal death, improvements in medical and obstetric care have decreased the likelihood of stillbirth. This article examines the current stillbirth rates reported in pregnancies complicated by common medical diseases. The reported stillbirth rates are expressed as the number of stillbirths occurring at > or = 20 weeks of gestation per 1,000 births in patients with the condition. Overall, about 10% of all fetal deaths are related to maternal medical illnesses such as hypertension, diabetes, obesity, systemic lupus erythematosus, chronic renal disease, thyroid disorders, and cholestasis of pregnancy. The early recognition of maternal medical diseases provides an opportunity for increased surveillance and interventions that may lead to more favorable pregnancy outcomes. PMID- 11876566 TI - Stillbirth: tissue findings with environmental and genetic links. AB - This article discusses stillbirth from a pathologist's perspective. Stillbirth may be caused by many different mechanisms. There are specific maternal and fetal disorders leading to stillbirth, and some of these have genetic and environmental associations. Frequently, an autopsy examination may determine the specific cause of the fetal death. We illustrate tissue findings in stillbirth to put individual faces on this condition. PMID- 11876568 TI - Inherited thrombophilia and stillbirth. AB - Thrombophilias are inherited or acquired conditions that predispose individuals to thromboembolism. New inherited thrombophilias are recognized each year. Some, but not all, studies have found an association between inherited thrombophilias and adverse pregnancy outcomes, including fetal loss. The controversy regarding the clinical implications of thrombophilias in pregnancy is clouded by differences in study populations, the number of thrombophilias tested, interactions between thrombophilias, and the retrospective nature of most studies, just to name a few factors. The lack of adequately designed studies also extends to clinical management. Clear evidence to determine when to test, whom to test, which thrombophilias to test for, when to treat, and what to treat with is not available. Further studies to investigate these questions are urgently needed. PMID- 11876569 TI - Genetics and metabolic causes of stillbirth. AB - Approximately 25% of stillbirths have been attributed to cytogenetic, mendelian, or biochemical causes with 75% still unknown. The most common autosomal trisomies are 21, 18, and 13, and the most common karyotypic abnormality is 45x. The remaining are sporadic multiple malformation syndromes and single organ malformations. Little is known about the genetic and metabolic causes of stillbirth; however, with new cytogenetic techniques such as fluorescent in situ hybridization, comparative genomic hybridization, and telomeric probes, cytogenetic errors will be identified more accurately. Advances in diagnosis will provide additional information for appropriate genetic counseling. PMID- 11876570 TI - The origins of stillbirth: infectious diseases. AB - This article reviews the literature on infectious diseases as a cause of stillbirth. Identifying a specific infection as a cause of stillbirth is limited by many obstacles. Nevertheless, "good faith" efforts estimate that approximately 9% to 15% of stillbirths are caused by infections. Infection may be especially important as a cause of stillbirth occurring early in pregnancy. Recognized causes include syphilis, toxoplasmosis, parvovirus B-19, chorioamnionitis, and Listeria monocytogenes. Other organisms that are "purported to cause" stillbirth include the genital mycoplasmas, Chlamydia trachomatis, HIV, group B streptococci, and others. No single strategy has been developed for effective prevention of stillbirth caused by infection. Six potential strategies and their rationale are presented, but the low rate of stillbirth in most situations would require large intervention trials for hypothesis testing. Infection is an important cause of stillbirth. Within the framework if determining epidemiological etiology, research should be initiated in the role of infectious causes. PMID- 11876571 TI - Umbilical cord accidents: human studies. AB - Research is needed to determine the cause of unexplained stillbirth. Sudden antenatal death syndrome is an important national issue that requires more scrutiny. Umbilical cord accidents as a causative factor of stillbirth need intensive investigation. Evidence supports a role of the umbilical cord in a portion of stillbirth cases, and theory suggests additional causes. This article summarizes the known information relating umbilical cord accidents and stillbirth and highlights the research needs. PMID- 11876572 TI - Neuropathology associated with stillbirth. AB - Neuropathologic findings in stillbirths oftentimes provide insight into the specific mechanisms leading to death. Examination of the brains of stillborn infants may also identify pathophysiologic processes that result in prenatal brain injury in liveborn as well as stillborn infants and that lead to neurologic disorders in liveborn infants, such as cerebral palsy or the sudden infant death syndrome (SIDS). A variety of abnormalities are found in the brains of stillborns, the most common including cerebral white matter necrosis (periventricular leukomalacia) or gliosis, germinal matrix or intraventricular hemorrhage, cerebral infarcts, pontosubicular necrosis, and spinal cord or brainstem necrosis. The 2 major hypotheses that have been proposed for the pathophysiology of cerebral white matter injury in the perinatal period are hypoxia/ischemia and infection/cytokines as the basis for injury. The fetal brain may be selectively vulnerable to various insults at specific stages of development. PMID- 11876574 TI - Contribution of efflux pump activity to the delivery of pulmonary therapeutics. AB - To date, there are few in vitro models of the human lung that have been used to characterize multidrug resistant (MDR) efflux pump activity. It is expected that the presence of these protein transporter molecules, such as P-glycoprotein (Pgp) and the multidrug resistance protein associated protein-1 (MRP1), might play a role in limiting drug absorption through the pulmonary epithelium, as has been reported for other epithelial drug delivery barriers such as the intestine and brain. To date, the exact role of the lung resistance related protein (LRP) in MDR is unclear. In this article, we have summarized the biochemistry, function and in vitro/in vivo modulation of Pgp and MRP1. These topics are discussed in light of pulmonary delivery of therapeutic agents, with particular emphasis being placed on the bronchial region of human airways. PMID- 11876573 TI - Sudden fetal and infant deaths: shared characteristics and distinctive features. AB - Recent evidence suggests sudden infant death syndrome (SIDS) infants have a diminished capacity to respond to autonomic challenges during a vulnerable developmental period. We speculate that a dysfunction or altered trajectory in the development of the autonomic nervous system may be detected in utero and also may play a role in the pathogenesis of unexplained late stillbirth. Some fetuses, as well as infants, may be more vulnerable than others to autonomic challenges during periods of autonomic instability. Investigation of potential shared underlying mechanisms in both SIDS and unexplained stillbirth will require expanded epidemiological investigation of genetic and environmental correlates along with a systematic study of developmental physiology and neuropathology. As with SIDS, there are likely important interactions between genetic susceptibility and environmental exposures occurring during gestation, which lead to infants who have altered trajectories or deficits in autonomic function and who need to be identified before they enter the periods of greatest risk. PMID- 11876575 TI - Selective serotonin reuptake inhibitors and cytochrome P-450 mediated drug-drug interactions: an update. AB - The selective serotonin reuptake inhibitors (SSRIs) have become the most prescribed antidepressants in many countries. Although the SSRIs share a common mechanism of action, they differ substantially in their chemical structure, metabolism, and pharmacokinetics. Perhaps the most important difference between the SSRIs is their potential to cause drug-drug interactions through inhibition of cytochrome-P450 (CYP) isoforms. This paper provides an update on both the in vitro and in vivo evidence with respect to CYP-mediated drug-drug interactions with this class of antidepressants. The available evidence clearly indicates that the individual SSRIs display a distinct profile of cytochrome P450 inhibition. Fluvoxamine is a potent CYP1A2 and CYP2C19 inhibitor, and a moderate CYP2C9, CYP2D6, and CYP3A4 inhibitor. Fluoxetine and paroxetine are potent CYP2D6 inhibitors, whereas fluoxetine's main metabolite, norfluoxetine, has a moderate inhibitory effect on CYP3A4. Sertraline is a moderate CYP2D6 inhibitor; citalopram appears to have little effect on the major CYP isoforms. Fluoxetine deserves special attention as inhibitory effects on CYP-activity can persist for several weeks after fluoxetine discontinuation because of the long half-life of fluoxetine and its metabolite norfluoxetine. Drug combinations with SSRIs should be assessed on an individual basis. Knowledge regarding the CYP-isoforms involved in the metabolism of the co-administered drug may help clinicians to anticipate and avoid potentially dangerous drug-drug interactions. Anticipated interactions can usually be managed by appropriate dose adjustment and titration of the object drug. In some cases, therapeutic drug monitoring can be useful. Equally well, an SSRI with limited interaction potential may be selected to treat depression in patients that receive other medications. PMID- 11876576 TI - Interaction of cyclosporine A and the renin-angiotensin system; new perspectives. AB - Despite extensive research, the exact mechanisms of cyclosporine A (CsA)-induced hypertension and nephrotoxicity remain obscure. Several lines of evidence suggest an involvement of the renin-angiotensin system (RAS) in CsA toxicity, but the issue is still controversial in more ways than one. Some interesting data of the interaction of CsA and RAS have been presented by us and others during the last years. In rats, activation of RAS by CsA is a consistent finding while the results from clinical studies show controversial results. The mechanisms of activation of RAS may be multifactorial. CsA increases renin release directly from juxtaglomerular cells. However, RAS activation may at least partly account for glomerular ischemia by vasoconstriction. A totally different view about the interaction of CsA and RAS has recently been presented. CsA antagonised the harmful effects of RAS over-expression on renal damage in double transgenic rats harbouring human renin and angiotensinogen genes. The protection was due to anti inflammatory properties of CsA by inhibition of interleukin-6 and inducible nitric oxide synthase (iNOS) expression. Other studies have confirmed the inhibitory effect of CsA on iNOS. Calcium antagonists have been proposed to be the antihypertensive drugs of choice in treatment of CsA-induced hypertension because of their favourable haemodynamic effects on the kidneys. However, because angiotensin II plays a major role in the development of CsA-induced structural renal damage, pharmacological inhibition of RAS in CsA-treatment may have some beneficial effects beyond blood pressure control. PMID- 11876577 TI - Combinations against combinations: associations of anti-HIV 1 reverse transcriptase drugs challenged by constellations of drug resistance mutations. AB - The reverse transcriptase inhibitors still represent the majority of the clinically used anti-HIV drugs and constitute the main backbone of currently employed combinatorial regimens. A major obstacle to successfull chemotherapic eradication of HIV is the emergence of viral strains resistant to the drugs in use. Counteracting the emergence of resistance necessitates alternating the panel of agents employed. In order to rationally design alternative drug combinations, physicians not only must know the genotype of the emerging viral strains, but should also be able to correlate it with its resistant phenotype. However, resistant viral strains usually carry multiple mutations, whose reciprocal influences on the overall level of resistance are largely unknown. Moreover, the choice of agents to be combined must take in account drug-drug interactions and adverse metabolic effects. This review will outline the main pharmacological and clinical features of the currently utilised anti-reverse transcriptase drugs, as well as the correspondent resistance profiles selected during therapy. A major focus will be on the reciprocal influence of drug associations on their own metabolism as well as on the interacting effects of the selected combinations of drug resistance mutations. PMID- 11876578 TI - The multidrug resistance mechanisms and their interactions with the radiopharmaceutical probes used for an in vivo detection. AB - The resistance of human malignancy to multiple chemotherapeutic agents ts remains a major obstacle in cancer therapy. This resistance phenomenon is called "multiple" because when cells are resistant they fail to respond to any of a wide range of anticancer agents. This leads to a complete ineffectiveness of any treatment and has dramatic consequences for the patients. This chemoresistance can be intrinsic--when tumour cells do not respond initially to the treatment--or acquired--when resistance appear during the therapy. Our understanding of the mechanisms responsible of the drug resistance has increased over the past few years. The tumour resistance is able to develop several strategies to inactivate the chemotherapeutic agents such as activation of the detoxification process, and overexpression of efflux pump proteins. The phenotype resistance of the cell is mainly characterised by an increased expression of membrane transport proteins such as the P-glycoprotein and the Multidrug Resistance Protein--MRPI--that act as real efflux pump to anticancer agent and contribute to physiological alterations i.e. intracellular pH and plasma membrane potentials. The detoxification procedure is also implicated with the Glutathione S transferase enzymes and the major anti oxidant of the cells the glutathione (GSH). More recently a newly reported transporter called "Breast Resistance Cancer Protein" has appeared. The role of all these transporters and the link with the detoxification systems in the clinical outcome of cancer chemotherapy is the subject of intense research. Particularly, one way of interest concerned in vivo investigations with radiolabelled compounds used in nuclear medicine. The understanding of how the radiolabelled compounds could interact with the phenotype resistance of the cells had a key role for further exploration of molecular imaging of the MDR phenotype. PMID- 11876579 TI - Similarities in the clinical characteristics related to alcohol dependence in two populations. AB - This report evaluates whether the characteristics associated with alcohol abuse and dependence are similar in two groups of men despite their enrollment in different research projects and the resulting differences in education and related background variables. Data regarding demographic and substance use characteristics and problems were gathered using similar research instruments from the 15-ear follow-up of 108 highly educated DSM-III-R alcohol-dependent men from the San Diego Prospective Study (Group 1) and 306 similarly diagnosed men with lower education from the Collaborative Study on the Genetics of Alcoholism (COGA ) (Group 2). Both groups of alcoholics reported high rates of alcohol relatedproblems, a similar maximum number of drinks per day, and similar proportions of drug use/non-use, although Group 1 subjects were less likely to report alcohol withdrawal and items associated with loss of control. Thus, teachers and clinicians may be able to generalize their knowledge and experience across different groups of alcoholics, using common sense modifications of their expectations based on the general characteristics of the groups involved despite differences in background variables, including education, income, marital status, and employment. PMID- 11876580 TI - Who starts treatment: engagement in the NIDA collaborative cocaine treatment study. AB - This study addressed the role of demographic variables, severity of drug dependence, and drug-related problems infacilitating or impeding engagement into a research treatment. Patients were tracked through various stages of entry for the research treatment: phone screening, intake, and randomization to treatment. Results suggested that certain demographic factors put patients more at risk for dropping out at intake and randomization. African-American and unemployed patients were less likely to stay in treatment at both stages, with largest differences at intake. Younger patients were less likely to complete both phases, with bigger differences noted at randomiZation. Patients with more days of cocaine use andpatients referred from advertisements were less likely to keep their intake appointments. PMID- 11876581 TI - Retention in psychosocial treatment of cocaine dependence: predictors and impact on outcome. AB - This report describes retention in treatment in the National Institute on Drug Abuse Collaborative Cocaine Treatment Study (CCTS), a multi-site trial of four psychosocial treatments for 487 cocaine dependent patients. Younger, African American, and unemployed patients were retained in treatment for fewer days than their counterparts. African-American patients who lived with a partner were retained in treatment for less time than if they lived alone. Higher psychiatric severity kept men in treatment longer but put women at risk for dropping out sooner. Patients who completed the full treatment used drugs less often than patients who dropped out, but outcome did not differ at each month. Patients in the drug counseling condition stayed in treatment for fewer days than patients in psychotherapy, but they were more likely to be abstinent after dropout. Patients with higher psychiatric severity were more at risk for continuing to use drugs after dropout. PMID- 11876582 TI - A family study of the high-risk children of opioid- and alcohol-dependent parents. AB - In this article, the author sought to use a high-risk study design focused on the children of parents with opioid or alcohol use disorders and children of parents with no substance use disorder (SUD) to evaluate the specificity of the risk conferred by the type of parental SUD. Using structured psychiatric interviews, cognitive assessments, and measures of social, academic, and family functioning, the authors studied 96 families (187 parents and 183 children, mean age of 11.6 years). Sixteen families had parental opioid dependence (22 children, 64% male), 14 families had parental alcohol dependence (22 children; 59% male), and 66 families had no SUD (controls; 139 children; 58% male). Fifty-nine percent of children of opioid-dependent parents had at least one major psychopathological condition, compared to 41% of the alcohol group and 28% of the control group (p < 0.01). The children of opioid- and alcohol-dependent parents were of lower socioeconomic status and had significantly more diffculties in academic, social, and family functioning than did controls. Children of opioid- and alcohol dependent parents have significantly higher rates of psychopathology as well as more difficulties in academic, social, and family functioning compared to the children of non-SUD parents. In addition, notable trends emerged for the opioid group to have more psychopathology and functional impairment than the alcohol group. With a growing consensus that certain risk factors for later SUD start in childhood with potentially treatable childhood-onset disorders, new preventive approaches for individuals at risk may be developed targeting childhood precursors of PMID- 11876583 TI - Outcomes of naltrexone maintenance following ultra rapid opiate detoxification versus intensive inpatient detoxification. AB - Relapse rates of 30 opiate-addicted social service clients who were given a 9 month course of naltrexone after being rapidly detoxified using naltrexone + clonidine under anesthesia were compared to 33 similar clients detoxified in a 30 day intensive inpatient detoxification but not given naltrexone. Both groups had the same counseling aftercare. Telephone follow-up of 26 of the intensive inpatient detoxification clients and 24 of the rapidly detoxified clients found no significant differences (p = .62) in relapse rates, with 34% of respondents returning to regular opiate use 13.4 (+/- 3.8) months after detoxification. Naltrexone maintenance and counseling following rapid detoxification may be as effective as intensive inpatient detoxification and counseling. PMID- 11876584 TI - The role of social support following short-term inpatient treatment. AB - The intensive, time-limited, short-term inpatient modality treatment for substance abuse appears to have positive outcomes despite its brevity. This study examined patient characteristics and posttreatment experiences to understand who is likely to benefit from this treatment and under what circumstances. Our sample included 748 patients in 12 short-term inpatient programs. Twenty-two percent of patients used cocaine at least weekly in the 1-year follow-up period, and an additional 9% drank frequently (compared with pretreatment rates of 69% and 15% respectively). Overall, patients' social support networks following treatment were more important factors than the pre- or during-treatment variables examined. PMID- 11876585 TI - The pharmacokinetics of methadone following co-administration with a lamivudine/zidovudine combination tablet in opiate-dependent subjects. AB - Methadone pharmacokinetics were determined in an open-label, within subject study in 16 methadone-maintained, non-HIV-infected subjects prior to and following administration of one lamivudine 150-mg/zidovudine 300-mg combination tablet to determine whether this antiretroviral therapy alters methadone serum concentrations. No significant differences in the mean area under the serum concentration-time curve (AUC(0-24h); 8,753 +/- 4,280 vs. 8,641 +/- 4,431 microg h/L),oralclearance(CL/F;9.9 +/- 4.9vs. 10.3 +/- 5.5 L/h),oral volume of distribution (Vd/F; 647 +/- 465 vs. 481 +/- 305 L), maximum serum concentration (Cmax; 514 +/- 223 vs. 5,510 +/- 237 microg/L), or terminal elimination half-life (t 1/2; 55.3 +/- 61.0 vs. 35.0 +/- 17.5 h) were detected. These results suggest that methadone dose change is not likely to be necessary for patients treated with lamivudine/zidovudine combination pharmacotherapy. PMID- 11876586 TI - Mental status changes in an alcohol abuser taking valerian and gingko biloba. PMID- 11876587 TI - Brain-derived neurotrophic factor and nitric oxide synthase inhibitor protect the vestibular organ against gentamicin ototoxicity. AB - In order to find a way to develop a new treatment for inner ear disorders, the effects of a nitric oxide synthase (NOS) inhibitor [N-nitro-L-arginine methylester (L-NAME)] and a neurotrophin [brain-derived neurotrophic factor (BDNF)] were investigated. The effect of L-NAME and BDNF on gentamicin-induced vestibular hair cell damage was investigated by using the in vitro LIVE/DEAD system. Both L-NAME and BDNF individually reduced the vestibular hair cell damage caused by gentamicin but the combination of L-NAME and BDNF was more successful in preventing damage. It is therefore suggested that treatment with a combination of an NOS inhibitor and a neurotrophin will help us to treat inner ear disorders. PMID- 11876588 TI - Enhanced production of matrix metalloproteinase-2 in human head and neck carcinomas is correlated with lymph node metastasis. AB - Matrix metalloproteinase (MMP)-2 and -9 degrade type IV collagen, which is one of the major components of the basement membrane in normal tissue and expressed in the surroundings of the cancer nest in squamous cell carcinoma. The degeneration of type IV collagen is an essential step in the metastasis to lymph nodes and distant organs. In this study, we examined MMP-2 and -9 levels of cancer tissue and serum obtained from patients with head and neck squamous cell carcinoma (HNSCC) in order to evaluate the relationship between the clinicopathologic features and MMPs. We examined the production of MMP-2 and -9 in cancer tissue homogenates of 73 patients who had HNSCC and the serum MMP levels of 16 patients with HNSCC and 8 healthy volunteers. We also studied the localization of MMP-2 in the carcinoma using an immunohistochemical approach. The concentrations of MMP-2 and -9 in the tissue homogenates and serum were measured by means of a sandwich enzyme immunoassay using a monoclonal antibody. Immunohistochemical analyses were performed with monoclonal antibody to MMP-2. The concentration of MMP-2 in the tumor tissue homogenates was unrelated to tumor size, but that in patients with lymph node metastases was significantly higher than in those without lymph node metastases. The concentration of MMP-9 was unrelated to lymph node metastasis and tumor size. The levels of both MMP-2 and -9 in serum were unrelated to lymph node metastasis. Immunohistochemistry indicated that MMP-2 was mainly expressed in cancer cells. Because MMP-2 degrades type IV collagen, the level of MMP-2 in carcinomas may be a useful indicator of the degree of invasion and metastasis. PMID- 11876589 TI - Occult cervical metastasis in head and neck cancer and its impact on therapy. PMID- 11876590 TI - Contemporary important considerations in diagnosis and treatment of head and neck cancer. PMID- 11876591 TI - Surgical management of recurrent tumor in the neck. PMID- 11876592 TI - Auditory behaviors and auditory brainstem responses of infants with hypogenesis of cerebral hemispheres. AB - Three infants with an almost complete absence of the cerebral hemispheres as a result of brain anomalies were studied both audiologically and neurologically. The brain anomalies were diagnosed by means of MRI and CT scans. Behavioral audiometry revealed reactions only to loud sound stimulations but auditory brainstem responses showed wave configurations and thresholds compatible with the ages of the infants. There were significant differences in the thresholds obtained by behavioral audiometry and auditory brainstem responses. It can be considered that these auditorily stimulated behavioral responses are evoked by auditory motor reflexes originating in the brainstem, but not by auditory perception. PMID- 11876593 TI - Sound-evoked postural responses in normal subjects. AB - A pattern of sound-induced paroxysms of the eye and head and other spinal motor neuron synkinesis (Tullio's phenomenon) in human subjects always implies either a pathological contiguity of the tympano-ossicular chain and membranous labyrinth or a dehiscence of the bone overlying the superior semicircular canal. However, it has become clear in the last decade that sound-evoked vestibular stimulation is not only a sign of disease but also a physiological phenomenon, The examination of such physiologically sound-induced vestibular (saccular) responses contributes today to the clinical testing of the vestibular organ, mainly in the form of vestibular-evoked myogenic potentials. In this study it was observed that, in a group of 20 normal subjects, a 500 Hz tonal stimulus of high intensity (105 dB HL = 118.5 dB SPL), applied monoaurally, elicited postural responses. Each subject was studied under 4 different conditions: (i) head facing forwards, eyes open; (ii) head facing forwards, eyes closed; (iii) head rotated approximately 90 degrees to the right, eyes closed: and (iv) head rotated approximately 90 degrees to the left, eyes closed. Body sway, measured using a force platform, was recorded in all subjects, with eyes either open or closed. Postural responses, which were also elicited with a 250 Hz tonal stimulus, were not observed with a tone of 2000 Hz, with legs slightly flexed or with binaural stimulation. The postural sway (head facing forwards, eyes open or closed) was in a lateral direction towards the stimulated ear: with the stimulus applied to the right ear the subject had postural sway towards the right, with the stimulus applied to the left ear towards the left. When the head was rotated approximately 90 degrees sideways and the stimulus was given facing forwards (i.e. head rotated contralaterally to stimulated ear) the postural sway was in a forward direction; when the head was rotated approximately 90 degrees sideways and the stimulus was given facing backwards (i.e. head rotated ipsilaterally to stimulated ear) the postural sway was in a backward direction. The mean values (mm) of body sway obtained with the head facing forwards and the eyes closed were higher than those with the eyes open (21.7 and 22.8 vs 15.7 and 14.7 for the right and left ears, respectively); higher mean values were obtained with the head turned to the side contralateral to the ear stimulated and the eyes closed (29.3 and 24.8 for the right and left ears, respectively). Under this condition the body sway was mainly in a forward direction. The sound-evoked vestibulopostural reflex seems to be a useful test for exploring the saccular function and, as a click-evoked vestibulocollic reflex, can be considered a physiological Tullio phenomenon. PMID- 11876594 TI - Treatment of anterior benign paroxysmal positional vertigo by canal plugging: a case report. AB - A 75-year-old man with incapacitating anterior canal benign paroxysmal positional vertigo (BPPV) was relieved of symptoms following anterior semicircular canal occlusion using a transmastoid approach. The preoperative symptoms were similar to those of posterior canal BPPV. The preoperative findings on Dix-Hallpike's maneuver were a paroxysmal torsional nystagmus with a down-beating component that increased when the patient's gaze was directed towards the affected ear. The most provoking head movement for the vertigo/nystagmus was Dix-Hallpike's maneuver with the affected ear lowermost. PMID- 11876595 TI - Validity and limitation of detection of peripheral vestibular imbalance from analysis of manually rotated vestibulo-ocular reflex recorded in the routine vestibular clinic. AB - We compared the results of analysis of vestibulo-ocular reflex (VOR) obtained by manual rotation in routine vestibular clinical practice with that of caloric testing, and examined the validity and limitations of VOR analysis as a test for the estimation of peripheral vestibular function and imbalance. VOR response was recorded in daily vestibular clinical examinations by manually rotating the standard clinical chair for approximately 30 s. VOR gain was slightly, but significantly, correlated with the peak slow phase velocity of caloric response (r = 0.50, p < 0.001). However, 8 out of 12 patients with no caloric response failed to exceed the range of two standard deviations of the mean value of age matched normal subjects, indicating that it is clinically difficult to use VOR gain alone as a estimate of unilateral vestibular function. VOR directional preponderance (VOR DP%) correlated well with caloric canal paresis (CP) (CP%; r = 0.89, p < 0.001). VOR DP% was within the normal range in patients with caloric CP% < 40 and exceeded the normal range in most cases with caloric CP% > 80. VOR DP% varied widely when caloric CP% ranged between 40 and 80. The effect of vestibular compensation on VOR DP% was examined by plotting VOR DP% divided by caloric CP% (DP/CP) against the number of days since the onset of vertigo in patients with vestibular neuritis or sudden deafness with vertigo. DP/CP was large within 50 days of the onset of vestibular damage, especially when caloric CP% was < 80, and gradually decreased with time. These results indicate that determination of VOR DP% should contribute to the early diagnosis of fresh vestibular imbalance, especially in daily clinical practice, because this type of VOR recording can be performed in < 1 min in routine vestibular clinics. The decay time constant of DP/CP was larger when caloric CP% exceeded 80, indicating that vestibular compensation proceeds more slowly when the vestibular damage is severe. PMID- 11876596 TI - Reverse optokinetic after-nystagmus generated by gaze fixation during optokinetic stimulation. AB - Gaze fixation during optokinetic stimulation generates an after-nystagmus with a slow component towards the reverse direction of the optokinetic stimulation. The duration and maximum slow component velocity (SCV) of this "reverse OKAN" were observed by changing the duration, velocity and direction of the optokinetic stimulation in nine normal volunteers. The duration of reverse OKAN increased with increasing stimulation time but was unaffected by changes in the stimulation velocity. The maximum SCV of reverse OKAN decreased with an increase in the stimulation velocity but was not significantly affected by changes in the optokinetic stimulation time. There was no directional difference among the horizontal, upwards and downwards reverse OKANs. The reverse OKAN was thought to be generated by a mechanism different from the velocity storage mechanism which produced optokinetic nystagmus and the first phase of OKAN. Retinal slip during the optokinetic stimulation was considered to be an input to the mechanism which generated the reverse OKAN. We hypothesize that the mechanism causing the reverse OKAN may be a generator of the second phase of OKAN, which was also intimately connected with self-motion sensation during the optokinetic stimulation. PMID- 11876597 TI - Dimensions of the cochlear nerve canal: a radioanatomic investigation. AB - The purpose of this study was to determine the normal variations of cochlear nerve canal dimensions, which is useful information to have when assessing congenital malformations, i.e. during the preoperative evaluation of cochlear implant candidates. The length and diameter of the cochlear nerve canal were measured in 117 casts from randomly selected temporal bone specimens obtained from the Uppsala Temporal Bone Laboratory. In 16 of the casts the dimensions were correlated with those obtained from CT scans of the same temporal bone. Measurements were also made from CT examinations of the temporal bone of 50 patients referred for evaluation of cholesteatoma or chronic otitis media. The mean length and diameter in the axiopetrosal plane measured in casts were 1.17 and 2.58 mm, respectively. The mean diameter in the axial plane was 2.59 mm. The mean length and diameter determined from CT scans of the specimens were 1.19 and 1.98 mm, respectively. The mean length and diameter determined from CT examinations of patients were 1.08 and 1.91 mm, respectively. In conclusion, the cochlear nerve canal is short, with a circular cross-section. If the diameter of the canal is < 1.4 mm then the possibility of cochlear nerve abnormality should be considered; if it is > 3.0 mm then other anomalies may coexist. PMID- 11876599 TI - Relationship between three inner ear antigens with different molecular weights and autoimmune inner ear disease. AB - Crude inner ear antigen (CIEAg) can induce autoimmune inner ear disease (AIED) although it is not known which subcomponent of CIEAg is involved. In this study, we investigated the relationship between 3 purified inner ear antigens (31, 42-45 and 60 kD proteins) and AIED, and determined their distribution in normal guinea pig cochlea. Three groups of guinea pigs were immunized with the three inner ear antigens and one group served as a control. The hearing thresholds, serum IgG level and morphological changes in the inner ear were observed. The expression of the three antigens in the cochlea was detected using immunohistochemical techniques. No obvious changes in hearing thresholds or inner ear morphology were observed between the control and 42-45 kD groups. Animals immunized with the 31 or 60 kD proteins showed a significant increase in hearing thresholds (p < 0.05 vs control), accompanied by morphological changes in the inner ear. The serum IgG level was increased significantly (p < 0.05) in all immunized animals. The 31 kD protein was distributed in the cochlear nerve and spiral ganglion, while the 42 45 and 60 kD proteins were distributed widely, being found in the spiral ganglion, organ of Corti, stria vascularis and spiral ligament. These results suggest that two subcomponents of CIEAg (the 31 and 60 kD proteins) may induce AIED independently, that several inner ear antigens may contribute to the pathogenesis of AIED and that the 31 kD protein is of high tissue specificity and may be used as a marker protein for the clinical diagnosis of AIED. PMID- 11876598 TI - Sensitivity of a new grading system for studying nasal polyps with the potential to detect early changes in polyp size after treatment with a topical corticosteroid (budesonide). AB - We have previously compared different scoring systems for endoscopic staging of nasal polyps. Of the five methods evaluated, we found that two were better than the others with regard to reproducibility and agreement between physicians. One method was lateral imaging, developed by the authors, and the other was a scoring system developed by Lildholdt et al. The main objective of the present study was to compare the sensitivity of these two methods. Another aim was to study the effect on nasal polyposis of topical nasal corticosteroids over a 2-week period. Patients with bilateral nasal polyposis (n = 100) were randomized to a 2-week treatment with a topical corticosteroid (budesonide aqueous nasal spray: 128 microg b.i.d.) or placebo in a double-blind manner. Nasal symptoms were scored before treatment and after 3, 7 and 14 days of treatment, and the patients underwent nasal endoscopy at clinical visits. Patients treated with active substance had an improvement in their symptoms, an effect already detectable after 3 days of treatment, compared with those who received placebo. In addition, a statistically significant decrease in polyp size could be registered after 14 days using lateral imaging but not with the other scoring system. In conclusion, lateral imaging was more sensitive and could detect effects earlier than the other scoring system and can be recommended for the endoscopic staging of nasal polyps in clinical studies. PMID- 11876600 TI - The effect of Japanese cedar-specific immunotherapy on cytokine production in peripheral blood mononuclear cells. AB - The most common cause of seasonal allergic rhinitis in Japan is the Japanese cedar (JC). Recently, a pullulan-conjugated antigen (CS-560) has been developed to reduce adverse effects and to enhance the effect of JC-specific immunotherapy (IT). If the mechanism of IT can be fully elucidated and the treatment can be used safely and with specificity, IT should be reconsidered as a superior treatment for JC pollinosis. Thirteen patients with JC pollinosis who received IT were compared to 10 patients who did not receive IT. All patients were followed through two pollen seasons by means of allergy diaries. Peripheral blood mononuclear cells (PBMC) were collected before IT and just before each pollen season, and these were stimulated with pollen extract. The concentrations of IL 4, IL-5, IL-13 and IFN-gamma in the culture supernatants were determined using an ELISA. Furthermore, messenger (m)RNA expressions of IL-4 and IL-5 from cultured PBMC were also studied. As a result of the allergy diaries, we confirmed the clinical efficacy of CS-560. The symptom-medication scores were significantly decreased by IT. The levels of IL-4, IL-5 and IL-13 declined only in the IT group. However, the level of IFN-gamma did not change in either group. IL-4 and IL-5 mRNA expressions were inhibited in the IT group compared to that in the non IT group. In conclusion, specific IT for JC pollinosis using CS-560 clearly modified cytokine expression by PBMC. PMID- 11876601 TI - Nasal and oral nitric oxide levels during experimental respiratory syncytial virus infection of adults. AB - Nitric oxide (NO) is a recognized mediator of inflammation in diseases affecting the lower respiratory tract, and has been implicated in the expression of viral upper respiratory tract infections. Here, exhaled nasal and oral NO concentrations and nitrite concentrations in nasal lavage fluids were measured, symptoms were scored and pulmonary function was evaluated before (Day 0) and after (Days 1-8) experimental exposure of 17 adult subjects to respiratory syncytial virus (RSV) type B. After exposure, RSV was recovered from 12 (70%) subjects by culture and/or specific antigen detection. Both subjects with and those without RSV recovery developed increased nasal and throat symptoms after RSV exposure, but none evidenced changes in the three measures of NO concentration. These results do not support the hypothesis of increased NO production during RSV infection and complement earlier studies that reported a lack of change in nasal NO concentration during experimental influenza and rhinovirus infections. PMID- 11876602 TI - Nasopharyngeal carriage of drug-resistant Streptococcus pneumoniae in children with acute otitis media evaluated by polymerase chain reaction-based genotyping of penicillin-binding proteins. AB - A polymerase chain reaction (PCR)-based genotyping of the penicillin-binding protein (PBP) genes pbp1a, pbp2x and pbp2b was used to characterize Streptococcus pneumoniae isolated from the nasopharynx of children with acute otitis media (AOM). Mutations were observed in pbp1a, pbp2x and pbp2b genes in 36.5% of the strains. Decreased susceptibility to beta-lactam antibiotics was closely associated with the frequency of mutations in the three PBP genes. Of penicillin intermediately-resistant S. pneumoniae strains, 54.5% appeared to be genetically similar to penicillin-resistant S. pneumoniae strains. Of penicillin-susceptible S. pneumoniae strains, 33.3% had mutations in the pbp2x gene and showed relatively high MICs to cephalosporins. Strains with mutations in the three PBP genes were often isolated from children < or = 2 years old. Evaluation of mutations in PBP genes using PCR will prove useful for studying the epidemiology of antibiotic resistance. PMID- 11876603 TI - Bacterial interference between pathogens in otitis media and alpha-haemolytic Streptococci analysed in an in vitro model. AB - Bacterial interference studied by means of agar methods has shown a decreased number of inhibitory alpha-haemolytic Streptococci among otitis-prone children. Additional information was gained regarding the interplay between alpha haemolytic Streptococci (AHS) and otitis media (OM) pathogens by comparing the bacterial interference in broth with the interference activity studied using agar overlay methods. We found, that non-typeable Haemophilus influenzae (NTHI) and Moraxella catarrhalis are readily inhibited by AHS in broth. Streptococcus pneumoniae was more bacteriostatically inhibited. If two OM pathogens were inoculated simultaneously, an isolate of AHS with poor inhibitory activity was not able to inhibit the growth, in contrast to an isolate of AHS with good inhibitory activity. The initial amount of AHS inoculated with M. catarrhalis seemed to play a decisive role with respect to the inhibitory activity. M. catarrhalis developed reduced susceptibility against AHS both in vivo and in vitro. In vivo studies showed that children with secretory otitis media had fewer isolates of AHS in their nasopharynx with the ability to inhibit all the test pathogens than healthy children (p < 0.001). Although the factor(s) responsible for the inhibitory activity have thus far not been defined, we could exclude low pH and nutrition depletion as the inhibitory mechanism of AHS with good inhibitory activity. PMID- 11876604 TI - Improvement in neuropsychological performance following surgical treatment for obstructive sleep apnea syndrome. AB - Obstructive sleep apnea syndrome (OSAS) is characterized by snoring and apnea during sleep leading to decreased oxygen saturation and disturbed sleep, excessive daytime sleepiness and neuropsychological disturbances. This study investigates cognitive neuropsychological abilities in a group of 53 OSAS patients before and after treatment with uvulopalatopharyngoplasty. General intellectual ability, verbal learning and memory as well as executive functioning were measured at baseline and 6 months postoperatively. After surgery there were significant improvements in verbal learning and memory (mean change -39, SD 57.3, p < 0.001), recall (mean change -24.3, SD 39.3, p < 0.001) and executive functioning (as assessed by percentage of errors on the Wisconsin Card Sorting Test; mean change -9.1, SD 15.7, p < 0.001). These improvements were in accordance with improvements in the degree of sleep apnea, the oxygen desaturation index (mean change -9.7, SD 15.9, p < 0.001) and arterial minimum oxygen saturation (mean change 4.5%, SD 10.2%, p < 0.01). Surgical treatment seems to improve verbal learning, memory and recall and executive functions in parallel with better oxygenation in OSAS. PMID- 11876605 TI - Intralaryngeal application of a miniaturized ultrasonic probe. AB - We developed a method for performing intralaryngeal ultrasonography. Normal larynges were obtained from 10 cadavers and examined using an intraluminal ultrasonic tomography apparatus connected to a radial scanning 20 MHz miniaturized probe. The larynx was placed in a bath filled with physiologic saline. and the probe was inserted through the forceps channel of the fiberscope (6 mm diameter). A horizontal ultrasonic image of the vocal fold was obtained. Histologic sections of the larynx were compared with the ultrasonic images. The mucosa in the membranous region of the vocal fold was comprised of three layers ultrasonographically. The epithelium and superficial layer of the lamina propria were visualized as a high echo (hyperechoic) region, the intermediate layer of the lamina propria was visualized as a low echo (hypoechoic) region and the deep layer of the lamina propria was seen as a hyperechoic region. The vocal fold structure can be visualized by intralaryngeal ultrasonography using the filling method. This method may be clinically useful for the detection of tumors involving the vocal folds. PMID- 11876606 TI - Juvenile nasopharyngeal angiofibroma: long-term results in preoperative embolized and non-embolized patients. AB - A treatment and follow-up study of 32 patients with juvenile nasopharyngeal angiofibroma (JNA) was performed at our clinic between 1974 and 1998. The majority had undergone surgery either via an antral approach or with a lateral rhinotomy. In the 1970s, surgery was combined with ligature of the external carotid artery and, since 1981, it has been combined with preoperative embolization. Two patients received radiotherapy (45 Gy) as primary treatment and the 3 cases of multiple recurrence received radiotherapy (30-45 Gy) as secondary treatment. No recurrence was found in patients treated with radiotherapy. The overall recurrence rate was 25%; the recurrence rate in non-embolized patients was 8% and among embolized patients it was 41%. We found no statistically verified differences in recurrence rate between embolized and non-embolized patients. No statistically significant difference was found in either recurrence or peroperative bleeding when comparing preoperatively embolized patients with non-embolized patients. Regression analyses showed that the only factor affecting recurrence was age, i.e. the younger the patient was at diagnosis the greater the risk of developing recurrence. The development of imaging and embolization techniques will hopefully contribute in the future towards reducing the recurrence rate. PMID- 11876607 TI - Use of Island Deltopectoral flap in reconstructive head neck oncology. AB - Deltopectoral flap is commonly used to reconstruct defects resulting from abalative head neck cancer surgery. Standard Deltopectoral flap has its own inherent technical drawbacks. Here with we describe a technique by which these are obviated. PMID- 11876608 TI - Primary papillary carcinoma in a thyroglossal duct cyst. AB - A rare case of primary thyroid papillary carcinoma arising in a thyroglossal duct cyst occuring in a 46 year old man is reported. The diagnosis was not suspected preoperatively. On gross examination of the excised specimen the presence of a papillary tumour in the cystic mass suggested the diagnosis which was proved histopathologically. The thyroid gland was normal on physical examination and scintigram thus ruling out the possibility of metastasis from a primary tumour in the thyroid gland. PMID- 11876609 TI - Identification of risk factors for specific subsites within the oral and oropharyngeal region--a study of 647 cancer patients. AB - Studies on site specific risks for oral cancers are few. Present investigation explores the possible role of human sociodemographic factors in causing oral cancer. Majority of patients had poor oral hygiene (85.5%) and belonged to 51-60 years age group (35.7%). Most of the subjects were agriculture workers (30.3%). Tongue and floor of mouth included majority of the affected sites (77.2%). Male to female ratio was highest for tonsil (32.3%) but differed marginally for other subsites. Majority of females used tobacco (81%) while males users of tobacco, alcohol and smoking reported in nearly equal proportions. Tobacco and smoking were found as primary risk factors for several intraoral subsites. However, for tongue, palate and lip no risk factor could be identified from given patients' characteristics. In general, tobacco posed high risk for buccal mucosa and alveolus in comparison to other subsites. Smoking affected tonsil and floor of mouth more than other sites. Alcohol posed more risk for buccal mucosa and floor of mouth than tongue. PMID- 11876610 TI - Central mucoepidermoid carcinoma--a case report. AB - The mucoepidermoid carcinoma (MEC) account for approximately 6-8 Percent of all salivary gland tumors. Central mucoepidermoid carcinoma of the jaws is rare. Following is a case report of central mucoepidermoid carcinoma which involves the ramus of the mandible. Origin of the central mucoepidermoid carcinoma is discussed. PMID- 11876611 TI - Prevention of carcinoma of cervix with human papillomavirus vaccine. AB - BACKGROUND: Carcinoma of cervix is the most common cancer found among the women of India. Though cervical cytology screening was effective in preventing carcinoma of cervix in developed nations, it is considered unsuitable in developing countries. Recent research has established an etiological link between human papillomavirus infection and carcinoma of cervix. In this review, an attempt is made to answer the question, 'whether carcinoma of cervix can be prevented with human papillomavirus vaccine?' METHODS: Literature search using Pubmed and Medline was carried out and relevant articles were reviewed. RESULTS: There is ample experimental evidence to show that DNA of human papillomavirus integrates with cervical cell genome. Viral genes E6 and E7 of HPV type 16 and 18 inactivate p53 function and Rb gene, thus immortalize the cervical epithelial cells. Recombinant vaccines blocked the function of E6 and E7 genes preventing development of papillomas in animals. Vaccination with HPV-VLPs encoding for genes of E6 and E7 neutralizes HPV integrated genome of malignant cells of uterine cervix. CONCLUSIONS: Based on experimental evidence, it is possible to prevent carcinoma of cervix with human papillomavirus vaccine, IMPLICATIONS: Further research is necessary to identify a effective and safe HPV vaccine, routes of administration and characteristics of potential beneficiaries. PMID- 11876612 TI - Supracricoid laryngectomy with Cricohyoidopexy--a clinico oncological & functional experience. AB - Supracricoid laryngectomy with Cricohyoidopexy (CHP) is a procedure that is commonly practiced in France & Canada. Eight such procedures were carried out at Kidwai Memorial Institute of Oncology, Bangalore during the period from 1991 through 1996. Four Glottic, 3 transglottic & one supraglottic cancers were subjected to this procedure. The study comprised of 7 males & 1 female. The average age was 52 years. Two procedures were done as salvage procedures for radiotherapy (RT) failures. The patients have a follow-up ranging from one year to six years, except for one who died soon after discharge from hospital secondary to myocardial infarction. Median follow up was four years. The three year acturial disease free survival was 83%. Six out of 8 (75%) were decannulated, and physiologic deglutition without aspiration was established in all patients. Hospital stay ranged from 11 to 62 days averaging 29 days. The speech was analyzed together with other partial laryngectomies and was found to be qualitatively worse than speech after other partial laryngectomy procedures. In addition speech intensity levels after CHP were lower than in other partial laryngectomy procedures. The speech however allowed normal social interaction. This procedure certainly has distinct oncological advantage in encompassing circumferential horse-shoe lesions with minimal subglottic extension which in the past would have received total laryngectomy and needs to be included in the repertoire of speech restorative surgery in laryngeal cancers. PMID- 11876613 TI - Evaluation of risk factors in CA CX at Government Medical College and Hospital, Aurangabad (Marathwada). AB - The Department of Radiotherapy & Oncology at Government Medical College, Aurangabad is a major cancer treatment centre of Marathwada region. The case records of 737 patients of carcinoma cervix registered from January 1996 to December 1997, have been reviewed and the various contributing etiological factors for the causation of cervical cancer relevant to this part of Maharashtra state, are analysed and an attempt has been made to identify high risk group. PMID- 11876614 TI - Histologic spectrum of struma ovarii. AB - Six cases of struma ovarii, including two with associated carcinoid, with unusual microscopic features are described. The patients' age ranged from 22 to 50 years. The patients were clinically euthyroid. The largest tumour was 10 cms. in great dimension. All tumours were unilateral. Microscopically various changes observed in a non-neoplastic thyroid gland were observed in the struma including Hashimotos thyroiditis and goitrous change. Adenomatous foci with microfollicles were observed in one case. A trabecular carcinoid admixed with mucinous glands lined bygoblet cells which were argyrophilic was seen in two of the neoplasms. This finding is rare and has been reported only once before. In conclusion it is important for a pathologist to be aware of the changes that may be associated with struma ovarii, so as not to confuse it with other neoplasms. A careful search for typical thyroid follicles, or other components of a dermoid help to arrive at a diagnosis. PMID- 11876615 TI - Acute Lympoblastic Leukaemia in seventy Iraqi adults: clinical and haematological findings and outcome of therapy. AB - Studies on acute Leukaemia from developing and Asian countries are scarce, and generally reflect poorer outcomes of therapy compared to their Western counterparts. This study was undertaken to address the latter issue in Iraqi adults with Acute Lymphoblastic Leukaemia (ALL). It included seventy unselected Iraqi adults (aged 14-60years), diagnosed as ALL in Baghdad Teaching Hospital, Baghdad, during the period between May 1991 and June 1994. The clinical and haematological findings in the included patients were generally comparable with those reported from the West, except for the lower median age. The patients were scheduled to receive a modified intensive chemotherapy protocol, and had an overall complete remission rate of 84.3%, and all overall median survival of 24 months. Nineteen patients were still alive in complete remission after a median follow-up of 67 months, and the estimated five year disease free survival was 27.2%. The above finding compare favourably with Western studies and are among the more favourable reports from Asian countries. The study also includes a discussion of the problems facing haematologists in the management of ALL in this part of the world. PMID- 11876616 TI - Malignant peritoneal mesothelioma in an inguinal hernial sac: an unusual presentation. AB - Malignant peritoneal mesothelioma, which is a rare neoplasm, usually presents with abdominal complaints. Though such tumours have been reported from tunica vaginalis testis presenting as para-testicular mass, there is only one documented case of the tumour arising from the inguinal hernial sac. In this paper, we are reporting a rare presentation of this tumour. PMID- 11876617 TI - Occurrence and severity of alopecia in patients on combination chemotherapy. AB - The aim of the study was to evaluate the occurrence and severity of alopecia resulting from combination chemotherapy on cancer patients. The study was conducted during the period 1994-1996 on 58 confirmed cases of malignancies attending the Kasturba Medical College Hospital, Mangalore, South India. The treatment regimens followed were standard protocols recommended for those malignancies and which are widely adopted. Specific drug combinations, their dosage and routes and schedules of administration were studied. The influence of 20 different treatment regimens, most of them in combination chemotherapy, were studied. The patients studied were not receiving any other medication which could have caused alopecia as observed in the present study. The pathophysiology of the hair, as influenced by the treatment regimens, were studied by examination of samples of the affected hairs under a Leica compound microscope. Alopecia was the most dominant side effect influencing 35 of the 58 patients undergoing the treatment (60%). The severity of alopecia was assessed by grouping them in four distinct grades. Specific drugs and their combinations causing varying degrees of severity were identified. The initiation of hair loss in different treatment regimens were analysed. It is seen that alopecia is an early manifestation of cutaneous side effects of cancer chemotherapy. In a majority of patients, the manifestation initiated after the first or the second cycle of administration of the rapeutic regimen, indicating a time interval of 1 to 8 weeks after the start of chemotherapy. Single agent drugs, when used alone or in combination with immunomodulator drugs seem to cause much less side effects, including alopecia, when compared to multiple drug regimens. Microscopic examination of the affected hair showed trichorrhexis, fragmentation, decrease in diameter and depigmentation of the hair shaft. PMID- 11876618 TI - Towards a more rational and scientific approach to vaccination. PMID- 11876619 TI - 2000 Report of the American Association of Feline Practitioners and Academy of Feline Medicine Advisory Panel on Feline Vaccines. PMID- 11876620 TI - Chylothorax in cats: is there a role for surgery? AB - Chylothorax is a complex disease with many identified underlying causes including cardiac disease, mediastinal masses, heartworm disease and trauma. Management of this disease should be directed at identifying the cause, if possible, and treating the underlying disorder. In cats with idiopathic chylothorax, medical management is recommended initially because the condition may resolve spontaneously. Owners should be made aware of the potential development of fibrosing pleuritis in affected cats. When medical management is impractical or unsuccessful, surgical intervention should be considered. Surgical options include mesenteric lymphangiography and thoracic duct ligation, pericardiectomy, omentalisation, passive pleuroperitoneal shunting, active pleuroperitoneal or pleurovenous shunting, and pleurodesis. Of these, only thoracic duct ligation and pericardiectomy are preferred by the author because, if successful, the result is complete resolution of the chylothorax, thereby reducing the risk of developing fibrosing pleuritis. Omentalisation may be beneficial in some animals as adjuvant therapy, but this procedure may still allow fibrosing pleuritis to occur. Until the aetiology of the effusion in cats with idiopathic chylothorax is understood, the treatment success rate will be less than ideal. Future research needs to be directed at determining the pathophysiologic mechanisms underlying this disease in cats. PMID- 11876621 TI - An investigation into the effects of storage on the diagnosis of crystalluria in cats. AB - Urinalysis was performed on 41 cats with no history of urinary tract disease. Samples were divided into aliquots, stored under differing condition and then examined for the presence of crystalluria. Crystalluria was detected in at least one stored sample in 92% of cats fed a mixed wet/dry food diet compared to 24% in the fresh sample. Crystalluria was not detected in any sample or aliquot from cats fed all wet food diets. PMID- 11876622 TI - Iatrogenic hyperadrenocorticism in a cat following a short therapeutic course of methylprednisolone acetate. AB - Iatrogenic hyperadrenocorticism (or iatrogenic Cushing's syndrome) is an adrenal disorder that may result from long-term administration of glucocorticoids for therapeutic purposes, most often given to treat allergic or immune-mediated disorders. Prolonged treatment with synthetic glucocorticoids can suppress hypothalamic corticotrophin releasing hormone and plasma adrenocorticotrophic hormone (ACTH), thus causing a functional inactivity of the adrenal cortex. The result is a clinical syndrome of hyperadrenocorticism but with basal and ACTH stimulated plasma cortisol concentrations that are consistent with spontaneous hypoadrenocorticism (Addison's disease). Whilst iatrogenic hyperadrenocorticism is relatively frequent in dogs, the diagnosis of iatrogenic hyperadrenocorticism in cats is very uncommon because this species has been found to be remarkably resistant to prolonged administration of glucocorticoids. To the author's knowledge, there are only two published clinical cases of feline iatrogenic Cushing's syndrome. This report describes a case of iatrogenic hyperadrenocorticism in a cat, and shows how normalisation of the adrenal function was achieved with supportive treatment and withdrawal of glucocorticoid administration. PMID- 11876623 TI - Feline acute pancreatitis--important species differences. PMID- 11876625 TI - Feline neuromuscular diseases. PMID- 11876624 TI - What is so special about feline diabetes mellitus? PMID- 11876626 TI - Genetic diseases of cats. PMID- 11876628 TI - Review of feline pancreatitis part one: the normal feline pancreas, the pathophysiology, classification, prevalence and aetiologies of pancreatitis. AB - The cellular mechanisms involved once pancreatitis has been initiated are reasonably well understood. The events leading up to this process are less well established. Much of our current understanding of pancreatitis in cats has been determined from experiments in cats or extrapolated from other species. The normal anatomy and function of the pancreas and a review of the current state of knowledge about the pathophysiology of pancreatitis is discussed. The current prevalence of feline pancreatitis is unknown, but the disease is being reported with increasing frequency. The aetiology of pancreatitis and the types of pancreatic inflammation present in cats is different from other species, such as the dog, a species where the disease is considered more common. Concurrent diseases are often present that may be more serious than the pancreatic inflammation and the treatment of these diseases is often complicated by pancreatitis. PMID- 11876629 TI - Review of feline pancreatitis part two: clinical signs, diagnosis and treatment. AB - In the past decade pancreatitis has become recognised as a significant disease in the cat. Chronic, mild pancreatitis is often associated with more commonly diagnosed diseases such as inflammatory bowel disease or cholangitis/cholangiohepatitis. Furthermore, acute pancreatitis with similar complications to those seen in dogs is now diagnosed more frequently in cats. Unfortunately, the clinical signs and clinicopathological findings in cats with pancreatitis are often non-specific and vague. The lack of specific signs often results in a diagnosis being made only when the veterinary surgeon has a strong index of suspicion for pancreatitis and vigorously pursues that diagnosis. Pancreatitis is an important disease in cats, has been implicated as a potential cause of diabetes mellitus, and when present complicates the treatment of diabetes and other intra-abdominal diseases in cats. PMID- 11876630 TI - Detection of feline leukaemia virus in blood and bone marrow of cats with varying suspicion of latent infection. AB - The purpose of this study was to determine if polymerase chain reaction (PCR) could be used to detect FeLV proviral DNA in bone marrow samples of cats with varying suspicion of latent infection. Blood and bone marrow samples from 50 cats and bone marrow from one fetus were collected, including 16 cats with diseases suspected to be FeLV-associated. Serum enzyme-linked immunosorbent assay (ELISA), blood and bone marrow immunofluorescent antibody test (IFA), and blood and bone marrow PCR were performed on each cat, and IFA and PCR on bone marrow of the fetus. Forty-one cats were FeLV negative. Five cats and one fetus were persistently infected with FeLV. Four cats had discordant test results. No cats were positive on bone marrow PCR only. It appears persistent or latent FeLV infection is not always present in conditions classically associated with FeLV. PMID- 11876631 TI - Clinical application of plasma clearance of iohexol on feline patients. AB - Glomerular filtration rate (GFR) was estimated by plasma clearance of iohexol (PCio) in 52 conscious cats presented for a variety of reasons to Angel Animal Hospital over a 2-year period. Cats were divided into four groups according to their clinical conditions and reasons for measuring PCio. The median PCio (ml/min/kg) was 3.68 in normal cats (NM), 2.39 in cats with suspected renal disease (SP), 1.35 in cats referred to confirm renal dysfunction (RD), and 0.84 in cats with apparent clinical signs of renal failure (RF). There was a significant difference between the results for each group. The respective medians of blood urea nitrogen (BUN) and plasma creatinine concentration (Pcr) (mg/dl) were 15 and 1.40 in NM cats, 21 and 1.71 in SP cats, 30 and 2.20 in RD cats, and 48 and 3.30 in RF cats. The reference values of BUN and Pcr were 21 +/- 7 mg/dl and 1.5 +/- 0.4 mg/dl (mean +/- SD). Diminished renal function could not be detected in SP cats by either BUN or Pcr, while a marked decrease of GFR was demonstrated before BUN and Pcr increased, indicating the insensitivity of BUN and Pcr in detecting renal dysfunction in cats. PCio can be performed non invasively in conscious cats, which improves the veterinarian's ability to detect early stages of chronic renal disease. PMID- 11876632 TI - Laboratory profiles in cats with different pathological and immunohistochemical findings due to feline infectious peritonitis (FIP). AB - Blood was collected from 55 cats with feline infectious peritonitis (FIP) and from 50 control cats in order to define whether differences in pathological findings and in distribution of feline coronaviruses (FCoV) can be associated with changes in haemograms, serum protein electrophoresis, and antibody titres. Compared to controls, the whole group of FIP-affected cats had blood changes consistent with FIP. Based on the pathological findings or on the immunohistochemical distribution of viral antigen, FIP-affected cats were divided in the following groups: subacute against acute lesions; low against strong intensity of positivity; intracellular against extracellular positivities; positive against negative lymph nodes. Lymphopenia was more evident in cats with acute forms, strong intensity of positivity, extracellular antigen and negative lymph nodes. Cats with positive lymph nodes had the most evident changes in the protein estimations. These results suggest that differences in pathological findings might depend on different reactive patterns to the FCoVs. PMID- 11876633 TI - Intervertebral disc extrusion in six cats. AB - Existing reports concerning intervertebral disc disease (IVDD) have focused almost exclusively on dogs, although a small number of individual case reports of IVDD of cats has been published. The medical records of six cats with IVDD were reviewed. Radiographic studies confirmed narrowed intervertebral disc spaces, mineralised intervertebral discs, and one or more extradural compressive lesions of the spinal cord in each cat. All disc extrusions were located in the thoracolumbar region. Surgical decompression of the spinal cord was achieved in all cats by means of hemilaminectomy and removal of compressive extradural material confirmed to be degenerative disc material. Good to excellent neurological recovery was noted in five of the six cats included in this report. Based on this review, it appears that IVDD of cats has many similarities to IVDD of dogs, and that healthy cats with acute intervertebral disc extrusion(s) respond favourably to surgical decompression of the spinal cord. PMID- 11876635 TI - Role of electrostatics in the interaction between cytochrome f and plastocyanin of the cyanobacterium Phormidium laminosum. AB - The role of charged residues on the surface of plastocyanin from the cyanobacterium Phormidium laminosum in the reaction with soluble cytochrome f in vitro was studied using site-directed mutagenesis. The charge on each of five residues on the eastern face of plastocyanin was neutralized and/or inverted, and the effect of the mutation on midpoint potentials was determined. The dependence of the overall rate constant of reaction, k(2), on ionic strength was investigated using stopped-flow spectrophotometry. Removing negative charges (D44A or D45A) accelerated the reaction and increased the dependence on ionic strength, whereas removing positive charges slowed it down. Two mutations (K46A, K53A) each almost completely abolished any influence of ionic strength on k(2), and three mutations (R93A, R93Q, R93E) each converted electrostatic attraction into repulsion. At low ionic strength, wild type and all mutants showed an inhibition which might be due to changes in the interaction radius as a consequence of ionic strength dependence of the Debye length or to effects on the rate constant of electron transfer, k(et). The study shows that the electrostatics of the interaction between plastocyanin and cytochrome f of P. laminosum in vitro are not optimized for k(2). Whereas electrostatics are the major contributor to k(2) in plants [Kannt, A., et al. (1996) Biochim. Biophys. Acta 1277, 115-126], this role is taken by nonpolar interactions in the cyanobacterium, leading to a remarkably high rate at infinite ionic strength (3.2 x 10(7) M(-1) s(-1)). PMID- 11876636 TI - Structure comparison of two conserved HNF-3/fkh proteins HFH-1 and genesis indicates the existence of folding differences in their complexes with a DNA binding sequence. AB - The hepatocyte nuclear factor 3 (HNF-3)/fork head (fkh) family contains a large number of transcription factors that recognize divergent DNA sequences via a winged helix binding motif. HNF-3/fkh proteins show a broad profile of DNA sequence-specificity in which one DNA sequence can be recognized by more than one HNF-3/fkh protein and each individual HNF-3/fkh protein has several DNA binding sequences. In this study, heteronuclear NMR methods were used to study the structures of the DNA binding domain of a conserved winged helix protein HFH-1 and its DNA complexes. The structural comparison of winged helix proteins HFH-1 and Genesis and their DNA complexes indicates that even two highly conserved HNF 3 family members can adopt different local structures when they contact an identical DNA binding sequence, while one of these two HNF-3 proteins seems to adopt only slightly different structures on different DNA binding sites. PMID- 11876637 TI - Solution structures of the cis- and trans-Pro30 isomers of a novel 38-residue toxin from the venom of Hadronyche Infensa sp. that contains a cystine-knot motif within its four disulfide bonds. AB - The primary sequence and three-dimensional structure of a novel peptide toxin isolated from the Australian funnel-web spider Hadronyche infensa sp. is reported. ACTX-Hi:OB4219 contains 38 amino acids, including eight-cysteine residues that form four disulfide bonds. The connectivities of these disulfide bonds were previously unknown but have been unambiguously determined in this study. Three of these disulfide bonds are arranged in an inhibitor cystine-knot (ICK) motif, which is observed in a range of other disulfide-rich peptide toxins. The motif incorporates an embedded ring in the structure formed by two of the disulfides and their connecting backbone segments penetrated by a third disulfide bond. Using NMR spectroscopy, we determined that despite the isolation of a single native homologous product by RP-HPLC, ACTX-Hi:OB4219 possesses two equally populated conformers in solution. These two conformers were determined to arise from cis/trans isomerization of the bond preceding Pro30. Full assignment of the NMR spectra for both conformers allowed for the calculation of their structures, revealing the presence of a triple-stranded antiparallel beta sheet consistent with the inhibitor cystine-knot (ICK) motif. PMID- 11876638 TI - The two-domain NK1 fragment of plasminogen: folding, ligand binding, and thermal stability profile. AB - The two-domain fragment N+K1 (rNK1) [Glu(1)-Glu(163)] of human plasminogen was expressed in E. coli as a hexahistidine-tagged fusion protein and chromatographically purified. The (1)H NMR spectrum supports proper folding of the K1 component within the refolded rNK1 construct (rNK1/K1). The functional properties of rNK1/K1 were investigated via intrinsic fluorescence titration with kringle-specific omega-aminocarboxylic acid ligands. The affinities closely match those previously measured for the isolated K1, which indicates that the N-domain does not significantly affect the interaction of ligands with the lysine binding site of K1. Far-UV CD spectra recorded for the N-domain suggest conformational plasticity and flexibility for the module. Two classes of spectra, referred to as types A and B, were identified with the type A spectrum reflecting a higher secondary structure content than that estimated for the type B spectrum. Subtracting the CD spectrum of rK1 from that of rNK1 yields a spectrum (Delta) which reflects the conformation of the N-domain within the rNK1 construct (rNK1/N). Delta resembles the type A spectrum, suggesting that rNK1/N adopts a relatively more ordered conformation, stabilized by the adjacent rNK1/K1 domain. In contrast, thermal unfolding curves determined via CD indicate that the rNK1/N slightly lowers the melting temperature (T(m)) of rNK1/K1. Independence of the two domains within rNK1 was tested by monitoring the thermal unfolding of rNK1/K1 when in the presence of the kringle-specific ligand AMCHA, which left the rNK1/N T(m) essentially unaffected, while increasing that of the rNK1/K1 by approximately 10 degrees C. PMID- 11876639 TI - Molecular basis and functional consequences of the dominant effects of the mutant band 3 on the structure of normal band 3 in Southeast Asian ovalocytosis. AB - Southeast Asian ovalocytosis (SAO) human red cell membranes contain similar proportions of normal band 3 and a mutant band 3 with a nine amino acid deletion (band 3 SAO). We employed specific chemical modification and proteolytic cleavage to probe the structures of band 3 in normal and SAO membranes. When the membranes were modified specifically at lysine residues with N-hydroxysulfosuccinimide-SS biotin, band 3 Lys-851 was not modified in normal membranes but quantitatively modified in SAO membranes. Normal and SAO membranes showed different patterns of band 3 proteolytic cleavage. Notably, many sites cleaved in normal membranes were not cleaved in SAO membranes, despite the presence of normal band 3 in these membranes. The mutant band 3 changes the structure of essentially all the normal band 3 present in the SAO membranes, and these changes extend throughout the normal band 3 molecules. The results also imply that band 3 in SAO membranes is present as hetero-tetramers or higher hetero-oligomers. The dominant structural effects of band 3 SAO on the other band 3 allele have important consequences on the functional and hematological properties of human red cells heterozygous for band 3 SAO. Analysis of the altered profile of biotinylation and protease cleavage sites suggests the location of exposed surfaces in the band 3 membrane domain and identifies likely interacting regions within the molecule. Our approach provides a sensitive method for studying structural changes in polytopic membrane proteins. PMID- 11876640 TI - Scaffolding functions of arrestin-2 revealed by crystal structure and mutagenesis. AB - Arrestin binding to activated, phosphorylated G protein-coupled receptors (GPCRs) represents a critical step in regulation of light- and hormone-dependent signaling. Nonvisual arrestins, such as arrestin-2, interact with multiple proteins for the purpose of propagating and terminating signaling events. Using a combination of X-ray crystallography, molecular modeling, mutagenesis, and binding analysis, we reveal structural features of arrestin-2 that may enable simultaneous binding to phosphorylated receptor, SH3 domains, phosphoinositides, and beta-adaptin. The structure of full-length arrestin-2 thus provides a uniquely oriented scaffold for assembly of multiple, diverse molecules involved in GPCR signal transduction. PMID- 11876641 TI - Diverse stability and catalytic properties of human tryptase alpha and beta isoforms are mediated by residue differences at the S1 pocket. AB - Recombinant human tryptases (rHTs) corresponding to alpha and beta isoforms were characterized. rHTbeta was similar to tryptase isolated from skin (HST); it was a tetramer, hydrolyzed model substrates efficiently, and was functionally unstable when incubated under physiological conditions. Activity was lost rapidly (t(1/2) approximately 1 min) by a reversible process similar to that observed for the spontaneous inactivation of HST. Circular dichroism (CD) and intrinsic fluorescence emission (IFE) spectra of active rHTbeta corresponded to those of active HST and upon spontaneous inactivation IFE decreased in parallel to activity loss. rHTalpha differed from HST in catalytic ability and stability. rHTalpha did not react with model substrates, an active site titrant, or a competitive inhibitor of HST/rHTbeta. IFE and CD spectra were similar to those of the active and not the spontaneously inactivated form of HST. Under physiological conditions, rHTalpha IFE decreased at a rate 900-fold slower than that observed for HST, and rHTalpha remained tetrameric when examined by size exclusion chromatography at physiological salt concentration. Thus, rHTalpha is a stable "inactive" form of HT. Three active site variants of rHTalpha, K192Q, D216G, and K192Q-D216G were characterized. Residues 192 and 216 (chymotrypsinogen numbers for residues 191 and 215 of rHTalpha) lie at the entrance to the primary specificity (S1) pocket, and the mutations converted them to the residues of HTbeta. While K192Q displayed the same properties as rHTalpha, the catalytic and stability characteristics of D216G and K192Q-D216G progressively approached those of HST. Thus, the contrasting stability/activity properties of rHTalpha and rHTbeta are largely related to differences at the S1 pocket. On the basis of the properties of the variants, we suggest that the side chain of Asp216 is blocking and stabilizing the S1 pocket and that this stabilization is sufficient to prevent spontaneous inactivation. PMID- 11876642 TI - Atomic resolution (0.98 A) structure of eosinophil-derived neurotoxin. AB - Human eosinophil-derived neurotoxin (EDN) is a small, basic protein that belongs to the ribonuclease A superfamily. EDN displays antiviral activity and causes the neurotoxic Gordon phenomenon when injected into rabbits. Although EDN and ribonuclease A have appreciable structural similarity and a conserved catalytic triad, their peripheral substrate-binding sites are not conserved. The crystal structure of recombinant EDN (rEDN) has been determined at 0.98 A resolution from data collected at a low temperature (100 K). We have refined the crystallographic model of the structure using anisotropic displacement parameters to a conventional R-factor of 0.116. This represents the highest resolution structure of rEDN determined to date and is only the second ribonuclease structure to be determined at a resolution greater than 1.0 A. The structure provides a detailed picture of the conformational freedom at the various subsites of rEDN, and the water structure accounts for more than 50% of the total solvent content of the unit cell. This information will be crucial for the design of tight-binding inhibitors to restrain the ribonucleolytic activity of rEDN. PMID- 11876643 TI - Toward an understanding of the role of dynamics on enzymatic catalysis in lactate dehydrogenase. AB - The motions of key residues at the substrate binding site of lactate dehydrogenase (LDH) were probed on the 10 ns to 10 ms time scale using laser induced temperature-jump relaxation spectroscopy employing both UV fluorescence and isotope-edited IR absorption spectroscopy as structural probes. The dynamics of the mobile loop, which closes over the active site and is important for catalysis and binding, were characterized by studies of the inhibitor oxamate binding to the LDH/NADH binary complex monitoring the changes in emission of bound NADH. The bound NAD-pyruvate adduct, whose pyruvate moiety likely interacts with the same residues that interact with pyruvate in its ternary complex with LDH, served as a probe for any relative motions of active site residues against the substrate. The frequencies of its C=O stretch and -COO(-) antisymmetric stretch shift substantially should any relative motion of the polar moieties at the active site (His-195, Asp-168, Arg-109, and Arg-171) occur. The dynamics associated with loop closure are observed to involve several steps with motions from 1 to 300 microms. Apart from the "melting" of a few residues on the protein's surface, no kinetics were observed on any time scale in experiments of the bound NAD-pyr adduct although the measurements were made with a high degree of accuracy, even for final temperatures close to the unfolding transition of the protein. This is contrary to simple physical considerations and models. These results show that, once a productive protein/substrate complex is formed, the binding pocket is very rigid with very little, if any, motion apart from the mobile loop. The results also show that loop opening involves concomitant movement of the substrate out of the binding pocket. PMID- 11876644 TI - Tissue factor alters the pK(a) values of catalytically important factor VIIa residues. AB - Blood coagulation is triggered when the serine protease factor VIIa (fVIIa) binds to cell surface tissue factor (TF) to form the active enzyme-cofactor complex. TF binding to fVIIa allosterically augments the enzymatic activity of fVIIa toward macromolecular substrates and small peptidyl substrates. The mechanism of this enhancement remains unclear. Our previous studies have indicated that soluble TF (sTF; residues 1-219) alters the pH dependence of fVIIa amidolytic activity (Neuenschwander et al. (1993) Thromb. Haemostasis 70, 970), indicating an effect of TF on critical ionizations within the fVIIa active center. The pKa values and identities of these ionizable groups are unknown. To gain additional insight into this effect, we have performed a detailed study of the pH dependence of fVIIa amidolytic activity. Kinetic constants of Chromozym t-PA (MeSO(2)-D-Phe-Gly-Arg pNA) hydrolysis at various pH values were determined for fVIIa alone and in complex with sTF. The pH dependence of both enzymes was adequately represented using a diprotic model. For fVIIa alone, two ionizations were observed in the free enzyme (pK(E1) = 7.46 and pK(E2) = 8.67), with at least a single ionization apparent in the Michaelis complex (pK(ES1) similar 7.62). For the fVIIa-TF complex, the pK(a) of one of the two important ionizations in the free enzyme was shifted to a more basic value (pK(E1) = 7.57 and pK(E2) = 9.27), and the ionization in the Michaelis complex was possibly shifted to a more acidic pH (pK(ES1) = 6.93). When these results are compared to those obtained for other well-studied serine proteases, K(E1) and K(ES1) are presumed to represent the ionization of the overall catalytic triad in the absence and presence of substrate, respectively, while K(E2) is presumed to represent ionization of the alpha-amino group of Ile(153). Taken together, these results would suggest that sTF binding to fVIIa alters the chemical environment of the fVIIa active site by protecting Ile(153) from deprotonation in the free enzyme while deprotecting the catalytic triad as a whole when in the Michaelis complex. PMID- 11876645 TI - Endoproteolysis of presenilin in vitro: inhibition by gamma-secretase inhibitors. AB - The final proteolytic step to generate the amyloid beta-protein (Abeta) of Alzheimer's disease (AD) from beta-amyloid precursor protein (APP) is achieved by presenilin (PS)-dependent gamma-secretase cleavage. AD-causing mutations in PS1 and PS2 result in a selective and significant increase in production of the more amyloidogenic Abeta42 peptide. PS1 and PS2 undergo endoproteolysis by an unknown enzyme termed presenilinase to generate the functional complex of N- and C terminal fragments (NTF/CTF). To investigate the endoproteolytic activity that generates active PS, we used a mammalian cell-free system that allows de novo human PS NTF and CTF generation. PS NTF and CTF generation in vitro was observed in endoplasmic reticulum (ER)-enriched fractions of membrane vesicles and to a lesser extent in Golgi/trans-Golgi-network (TGN)-enriched fractions. AD-causing mutations in PS1 and PS2 did not alter de novo generation of PS fragments. Removal of peripheral membrane-associated and cytosolic proteins did not prevent de novo generation of fragments, indicating that presenilinase activity corresponds to an integral membrane protein. Among several general inhibitors of different protease classes that blocked the presenilinase activity, pepstatin A was the most potent inhibitor. Screening available transition state analogue gamma-secretase inhibitors led to the identification of two compounds that were able to prevent the de novo generation of PS fragments, with an expected inhibition of Abeta generation. Our studies provide a biochemical approach to characterize and identify this elusive presenilinase. PMID- 11876646 TI - Topology of the anion exchange protein AE1: the controversial sidedness of lysine 743. AB - The topology of the band 3 (AE1) polypeptide of the erythrocyte membrane is not fully established despite extensive study. Residues near lysine 743 (K743) have been reported to be extracellular in some studies and cytoplasmic in others. In the work presented here, we have attempted to establish the sidedness of K743 using in situ proteolysis. Trypsin, papain, and proteinase K do not cleave band 3 at or near K743 in intact red cells, even under conditions that cause cleavage on the C-terminal side of the glycosylation site (N642) in extracellular loop 4. In contrast, trypsin sealed inside red cell ghosts cleaves at K743, as does trypsin treatment of inside-out vesicles (IOVs). The transport inhibitor 4,4' diisothiocyanatodihydrostilbene-2,2'-disulfonate (H(2)DIDS), acting from the extracellular side, blocks trypsin cleavage at K743 in unsealed membranes by inducing a protease-resistant conformation. H(2)DIDS added to IOVs does not prevent cleavage at K743; therefore, trypsin cleavage at K743 in IOVs is not a consequence of cleavage of right-side-out or leaky vesicles. Finally, microsomes were prepared from HEK293 cells expressing the membrane domain of AE1 lacking the normal glycosylation site. This polypeptide does not traffic to the surface membrane; trypsin treatment of microsomes containing this polypeptide produces the 20 kDa fragment, providing further evidence that K743 is exposed at the cytoplasmic surface. Therefore, the actions of trypsin on intact cells, resealed ghosts, unsealed ghosts, inside-out vesicles, and microsomes from HEK293 cells all indicate that K743 is cytoplasmic and not extracellular. PMID- 11876647 TI - Amyloid fibril formation of the mouse V(L) domain at acidic pH. AB - The recombinant V(L) domain that represents the variable part of the light chain (type kappa) of mouse monoclonal antibody F11 directed against human spleen ferritin was found to form amyloid fibrils at acidic pH as evidenced by electron microscopy, thioflavin T binding, and apple-green birefringence after Congo red staining. This is the first demonstration of amyloid fibril formation of the mouse V(L) domain. To understand the mechanism of acidic pH-induced amyloid fibril formation, conformational changes of the V(L) domain were studied by one dimensional NMR, differential scanning calorimetry, analytical ultracentrifugation, hydrophobic dye binding, far-UV circular dichroism, and tryptophan fluorescence. The results indicated accumulation of two intermediate states during acid unfolding, which might be responsible for amyloid fibril formation. The more structured intermediate that exhibited maximal accumulation at pH 3 retained the nativelike secondary structure and a hydrophobic core, but exposed hydrophobic surfaces that bind 8-anilino-1-naphthalenesulfonate. Below pH 2, a more disordered intermediate with dequenched tryptophan fluorescence but still retaining the beta-sheet structure accumulated. The optimal pH of amyloid fibril formation (i.e., pH 4) was close to the optimal pH of the accumulation of the nativelike intermediate, suggesting that the amyloid fibrils might be formed through this intermediate. PMID- 11876648 TI - Nitric oxide activation of soluble guanylyl cyclase reveals high and low affinity sites that mediate allosteric inhibition by calcium. AB - Cyclic GMP (cGMP) and Ca(2+) regulate opposing mechanisms in (patho)physiological processes reflected in the reciprocal regulation of their intracellular concentrations. Although mechanisms by which cGMP regulates [Ca(2+)](i) have been described, those by which Ca(2+) regulates [cGMP](i) are less well understood. In the present study, Ca(2+) inhibited purified sGC activated by sodium nitroprusside (SNP), a precursor of nitric oxide (NO), employing Mg-GTP as substrate in a concentration-dependent fashion, but was without effect on basal enzyme activity. Ca(2+) inhibited sGC stimulated by protoporphyrin IX or YC-1 suggesting that inhibition was not NO-dependent. In contrast, Ca(2+) was without effect on sGC activated by SNP employing Mn-GTP as substrate, demonstrating that inhibition did not reflect displacement of heme from sGC. Ligand activation of sGC unmasked negative allosteric sites of high (K(i) similar 10(-7) M) and low (K(i) approximately 10(-5) M) affinity for Ca(2+) that mediated noncompetitive and uncompetitive inhibition, respectively. Free Mg(2+) in excess of substrate did not alter the concentration-response relationship of Ca(2+) inhibition at high affinity sites, but produced a rightward shift in that relationship at low affinity sites. Similarly, Ca(2+) inhibition at high affinity sites was noncompetitive, whereas inhibition at low affinity sites was competitive, with respect to free Mg(2+). Purified sGC specifically bound (45)Ca(2+) in the presence of a 1000-fold excess of Mg(2+) and in the absence of activating ligands. These data suggest that sGC is a constitutive Ca(2+) binding protein whose allosteric function is conditionally dependent upon ligand activation. PMID- 11876649 TI - The low lysine content of ricin A chain reduces the risk of proteolytic degradation after translocation from the endoplasmic reticulum to the cytosol. AB - Several protein toxins, including the A chain of ricin (RTA), enter mammalian cells by endocytosis and subsequently reach their cytosolic substrates by translocation across the endoplasmic reticulum (ER) membrane. To achieve this export, such toxins exploit the ER-associated protein degradation (ERAD) pathway but must escape, at least in part, the normal degradative fate of ERAD substrates. Toxins that translocate from the ER have an unusually low lysine content. Since lysyl residues are potential ubiquitination sites, it has been proposed that this paucity of lysines reduces the chance of ubiquitination and subsequent ubiquitin-mediated proteasomal degradation [Hazes, B., and Read, R. J. (1997) Biochemistry 36, 11051-11054]. Here we provide experimental support for this hypothesis. The two lysyl residues within RTA were changed to arginyl residues. Their replacement in RTA did not have a significant stabilizing effect, suggesting that the endogenous lysyl residues are not the usual sites for ubiquitin attachment. However, when four additional lysines were introduced into RTA in a way that did not compromise the activity, structure, or stability of the toxin, degradation was significantly enhanced. Enhanced degradation resulted from ubiquitination that predisposed the toxin to proteasomal degradation. Treatment with the proteasome inhibitor clasto-lactacystin beta-lactone increased the cytotoxicity of the lysine-rich RTA to a level approaching that of wild-type ricin. The introduction of four additional lysyl residues into a second ribosome inactivating protein, abrin A chain, also dramatically decreased the cytotoxicity of the holotoxin compared to wild-type abrin. This effect could also be reversed by proteasomal inhibition. Our data support the hypothesis that the evolution of a low lysine content is a degradation-avoidance strategy for toxins that retrotranslocate from the ER. PMID- 11876650 TI - Phospholipase D2 directly interacts with aldolase via Its PH domain. AB - Mammalian phospholipase D (PLD) has been implicated in the cellular signal transduction pathways leading to diverse physiological events and known to be regulated by many cellular factors. To identify the proteins that interact with PLD, we performed a protein overlay assay with fractions obtained from the sequential column chromatographic separation of rat brain cytosol using purified PLD2 as a probe. A protein of molecular mass 40 kDa, which was detected by anti PLD antibody with overlaying of the purified PLD2, is shown to be aldolase C by peptide-mass fingerprinting with matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF-MS). Aldolase A also showed similar binding properties as aldolase C and was co-immunoprecipitated with PLD2 in COS-7 cells overexpressing PLD2 and aldolase A. The PH domain corresponding to amino acids 201-310 of PLD2 was necessary for the interaction observed in vitro, and aldolase A was found to interact with the PH domain of PLD2 specifically, but not with other PH domains. PLD2 activity was inhibited by the presence of purified aldolase A in a dose-dependent manner, and the inhibition by 50% was observed by the addition of less than micromolar aldolase A. Moreover, the inclusion of the aldolase metabolites fructose 1,6-bisphosphate (F-1,6-P) or glyceraldehyde 3 phosphate (G-3-P) resulted in an enhanced interaction between PLD2 and aldolase A with a concomitant increase in the potential ability of aldolase A to inhibit PLD2, which suggests the existence of a possible regulation of the interaction by the change of intracellular concentrations of glycolytic metabolites. PMID- 11876651 TI - Electron transport routes in whole cells of Synechocystis sp. strain PCC 6803: the role of the cytochrome bd-type oxidase. AB - The plastoquinone pool is the central switching point of both respiratory and photosynthetic electron transport in cyanobacteria. Its redox state can be monitored noninvasively in whole cells using chlorophyll fluorescence induction, avoiding possible artifacts associated with thylakoid membrane preparations. This method was applied to cells of Synechocystis sp. PCC 6803 to study respiratory reactions involving the plastoquinone pool. The role of the respiratory oxidases known from the genomic sequence of Synechocystis sp. PCC 6803 was investigated by a combined strategy using inhibitors and deletion strains that lack one or more of these oxidases. The putative quinol oxidase of the cytochrome bd-type was shown to participate in electron transport in thylakoid membranes. The activity of this enzyme in thylakoids was strongly dependent on culture conditions; it was increased under conditions where the activity of the cytochrome b(6)f complex alone may be insufficient for preventing over-reduction of the PQ pool. In contrast, no indication of quinol oxidase activity in thylakoids was found for a second alternative oxidase encoded by the ctaII genes. PMID- 11876652 TI - Alcaligenes xylosoxidans dissimilatory nitrite reductase: alanine substitution of the surface-exposed histidine 139l ligand of the type 1 copper center prevents electron transfer to the catalytic center. AB - Nitrite reductase of Alcaligenes xylosoxidans contains three blue type 1 copper centers with a function in electron transfer and three catalytic type 2 copper centers. The mutation H139A, in which the solvent-exposed histidine ligand of the type 1 copper ion was changed to alanine, resulted in the formation of a colorless protein containing 4.4 Cu atoms per trimer. The enzyme was inactive with reduced azurin as the electron donor, and in contrast to the wild-type enzyme, no EPR features assignable to type 1 copper centers were observed. Instead, the EPR spectrum of the H139A enzyme, with parameters of g(1) = 2.347 and A(1) = 10 mT, was typical of type 2 copper centers. On the addition of nitrite, the EPR features developed spectral features with increased rhombicity, with g(1) = 2.29 and A(1) = 11 mT, arising from the type 2 catalytic site. As assessed by visible spectroscopy, ferricyanide (E degree = +430 mV) was unable to oxidize the H139A enzyme, and this required a 30-fold excess of K(2)IrCl(6) (E degree = +867 mV). Oxidation resulted in the EPR spectrum developing additional axial features with g(1) = 2.20 and A(1) = 9.5 mT, typical of type 1 copper centers. The oxidized enzyme after separation from the excess of K(2)IrCl(6) by gel filtration was a blue-green color with absorbance maxima at 618 and 420 nm. The instability of the protein prevented the precise determination of the midpoint potential, but these properties indicate that it is in the range 700-800 mV, an increase of at least approximately 470 mV compared with the native enzyme. This high potential, which is consistent with a trigonal planar geometry of the Cu ion, effectively prevents azurin-mediated electron transfer from the type 1 center to the catalytic type 2 Cu site. However, with dithionite as reductant, 20% of the activity of the wild-type enzyme was observed, indicating that the direct reduction of the catalytic site by dithionite can occur. When CuSO(4) was added to the crude extract before isolation of the enzyme, the Cu content of the purified H139A enzyme increased to 5.7 Cu atoms per trimer. The enzyme remained colorless, and the activity with dithionite as a donor was not significantly increased. The additional copper in such preparations was associated with an axial type 2 Cu EPR signal with g(1) = 2.226 and A(1) = 18 mT, and which were not changed by the addition of nitrite, consistent with the activity data. PMID- 11876653 TI - Reactions catalyzed by the heme domain of inducible nitric oxide synthase: evidence for the involvement of tetrahydrobiopterin in electron transfer. AB - The heme domain (iNOS(heme)) of inducible nitric oxide synthase (iNOS) was expressed in Escherichia coli and purified to homogeneity. Characterization of the expressed iNOS(heme) shows it to behave in all respects like full-length iNOS. iNOS(heme) is isolated without bound pterin but can be readily reconstituted with (6R)-5,6,7,8-tetrahydro-L-biopterin (H(4)B) or other pterins. The reactivity of pterin-bound and pterin-free iNOS(heme) was examined, using sodium dithionite as the reductant. H(4)B-bound iNOS(heme) catalyzes both steps of the NOS reaction, hydroxylating arginine to N(G)-hydroxy-L-arginine (NHA) and oxidizing NHA to citrulline and *NO. Maximal product formation (0.93 plus minus 0.12 equiv of NHA from arginine and 0.83 plus minus 0.08 equiv of citrulline from NHA) requires the addition of 2 to 2.5 electron equiv. Full reduction of H(4)B bound iNOS(heme) with dithionite also requires 2 to 2.5 electron equiv. These data together demonstrate that fully reduced H(4)B-bound iNOS(heme) is able to catalyze the formation of 1 equiv of product in the absence of electrons from dithionite. Arginine hydroxylation requires the presence of a bound, redox-active tetrahydropterin; pterin-free iNOS(heme) or iNOS(heme) reconstituted with a redox inactive analogue, 6(R,S)-methyl-5-deaza-5,6,7,8-tetrahydropterin, did not form NHA under these conditions. H(4)B has an integral role in NHA oxidation as well. Pterin-free iNOS(heme) oxidizes NHA to citrulline, N(delta)-cyanoornithine, an unidentified amino acid, and NO(-). Maximal product formation (0.75 plus minus 0.01 equiv of amino acid products) requires the addition of 2 to 2.5 electron equiv, but reduction of pterin-free iNOS(heme) requires only 1 to 1.5 electron equiv, indicating that both electrons for the oxidation of NHA by pterin-free iNOS(heme) are derived from dithionite. These data provide strong evidence that H(4)B is involved in electron transfer in NOS catalysis. PMID- 11876654 TI - Potent gene-specific inhibitory properties of mixed-backbone antisense oligonucleotides comprised of 2'-deoxy-2'-fluoro-D-arabinose and 2'-deoxyribose nucleotides. AB - Phosphorothioate deoxyribonucleotides (PS-DNA) are among the most widely used antisense inhibitors. PS-DNA exhibits desirable properties such as enhanced nuclease resistance, improved bioavailability, and the ability to induce RNase H mediated degradation of target RNA. Unfortunately, PS-DNA possesses a relatively low binding affinity for target RNA that impacts on its potency in antisense applications. We recently showed that phosphodiester-linked oligonucleotides comprised of 2'-deoxy-2'-fluoro-D-arabinonucleic acid (FANA) exhibit both high binding affinity for target RNA and the ability to elicit RNase H degradation of target RNA [Damha et al. (1998) J. Am. Chem. Soc. 120, 12976]. In the present study, we evaluated the antisense activity of phosphorothioate-linked FANA oligonucleotides (PS-FANA). Oligonucleotides comprised entirely of PS-FANA were somewhat less efficient in directing RNase H cleavage of target RNA as compared to their phosphorothioate-linked DNA counterparts, and showed only weak antisense inhibition of cellular target expression. However, mixed-backbone oligomers comprised of PS-FANA flanking a central core of PS-DNA were found to possess potent antisense activity, inhibiting specific cellular gene expression with EC(50) values of less than 5 nM. This inhibition was a true antisense effect, as indicated by the dose-dependent decrease in both target protein and target mRNA. Furthermore, the appearance of mRNA fragments was consistent with RNase H mediated cleavage of the mRNA target. We also compared a series of PS-[FANA-DNA FANA] mixed-backbone oligomers of varying PS-DNA core sizes with the corresponding 2'-O-methyl oligonucleotide chimeras, i.e., PS-[2'meRNA-DNA 2'meRNA]. Both types of oligomers showed very similar binding affinities toward target RNA. However, the antisense potency of the 2'-O-methyl chimeric compounds was dramatically attenuated with decreasing DNA core size, whereas that of the 2' fluoroarabino compounds was essentially unaffected. Indeed, a PS-FANA oligomer containing a single deoxyribonucleotide residue core retained significant antisense activity. These findings correlated exactly with the ability of the various chimeric antisense molecules to elicit RNase H degradation of the target RNA in vitro, and suggest that this mode of inhibition is likely the most important determinant for potent antisense activity. PMID- 11876655 TI - Fluorescence studies of pyrene maleimide-labeled translin: excimer fluorescence indicates subunits associate in a tail-to-tail configuration to form octamer. AB - Translin is an octameric single-stranded DNA binding protein consisting of 228 amino acid residues per monomer. This protein contains two cysteine residues per monomer. Studies of reactions with DTNB show that both cysteines are reactive and exhibit biphasic reaction kinetics. Further studies with two site-directed mutants, C58S and C225S, confirm that Cys-58 reacts slowly while Cys-225 reacts quickly. Pyrene excimer emission was observed for pyrene maleimide-labeled C58S mutant. This was not observed, however, with the pyrene maleimide-labeled C225S mutant. DAS (decay associated spectra) revealed that all excited pyrene labels on C225 residues can form excimers with pyrenes of adjacent subunits within a few nanoseconds. Time-resolved emission anisotropy detects a rotational correlation time appropriate for octameric but not dimeric species. These results indicate proximity for the Cys-225 residues on adjacent monomers and that the subunits must interact in a tail-to-tail orientation. Moreover, disulfide bonds are not required for the formation of an octamer. PMID- 11876656 TI - cis-Retinol/androgen dehydrogenase, isozyme 3 (CRAD3): a short-chain dehydrogenase active in a reconstituted path of 9-cis-retinoic acid biosynthesis in intact cells. AB - 9-cis-Retinoic acid activates retinoid X receptors, which serve as heterodimeric partners with other nuclear hormone receptors, yet the enzymology of its physiological generation remains unclear. Here, we report the identification and molecular/enzymatic characterization of a previously unknown member of the short chain dehydrogenase/reductase family, CRAD3 (cis-retinoid/androgen dehydrogenase, type 3), which catalyzes the first step in 9-cis-retinoic acid biosynthesis, the conversion of 9-cis-retinol into 9-cis-retinal. CRAD3 shares amino acid similarity with other retinoid/steroid short-chain dehydrogenases/reductases: CRAD1, CRAD2, and RDH4. Relative to CRAD1, CRAD3 has greater 9-cis-retinol/all trans-retinol discrimination and lower efficiency as an androgen dehydrogenase. CRAD3 has apparent efficiency (V/K(m)) for 9-cis-retinol about equivalent to that for CRAD1 and 3 orders of magnitude greater than that for RDH4. (CRAD2 does not recognize 9-cis-retinol as a substrate). CRAD3 contributes to 9-cis-retinoic acid production in intact cells, in conjunction with each of three retinal dehydrogenases that recognize 9-cis-retinal (RALDH1/AHD2, RALDH2, and ALDH12). Liver and kidney, two tissues reportedly with the highest concentrations of 9-cis retinoids, show the most intense mRNA expression of CRAD3, but expression also occurs in testis, lung, small intestine, heart, and brain. These data are consistent with the participation of CRAD3 in the biogeneration of 9-cis-retinoic acid. PMID- 11876657 TI - Interaction of Hoechst 33258 and echinomycin with nucleosomal DNA fragments containing isolated ligand binding sites. AB - We have examined the interaction of Hoechst 33258 and echinomycin with nucleosomal DNA fragments which contain isolated ligand binding sites. A 145 base pair fragment was prepared on the basis of the sequence of tyrT DNA, which contained no CpG or (A/T)(4) binding sites for these ligands. Isolated binding sites were introduced into this fragment at discrete locations where the minor groove is known to face toward or away from the protein core when reconstituted onto nucleosome core particles. The interaction of ligands with target sites on these nucleosomal DNA fragments was assessed by DNase I footprinting. We find that Hoechst 33258 can bind to single nucleosomal sites which face both toward and away from the protein core, without affecting the nucleosome structure. Hoechst binding is also observed on nucleosomal fragments which contain two or more drug binding sites, though in these cases the footprints are accompanied by the presence of new cleavage products in positions which suggest that the ligand has caused a proportion of the DNA molecules to adopt a new rotational positioning on the protein surface. Hoechst 33258 does not affect nucleosome reconstitution with any of these fragments. In contrast, the bifunctional intercalating antibiotic echinomycin is not able to bind to single nucleosomal CpG sites. Echinomycin footprints are observed on nucleosomal fragments containing two or more CpG sites, but there are no changes in the cleavage patterns in the remainder of the fragment. Echinomycin abolishes nucleosome reconstitution when included in the reconstitution mixture. PMID- 11876658 TI - Lignin peroxidase oxidation of veratryl alcohol: effects of the mutants H82A, Q222A, W171A, and F267L. AB - The site-directed mutations H82A and Q222A (residues near the heme access channel), and W171A and F267L (residues near the surface of the protein) were introduced into the gene encoding lignin peroxidase (LiP) isozyme H8 from Phanerochaete chrysosporium. The variant enzymes were produced by homologous expression in P. chrysosporium, purified to homogeneity, and characterized by kinetic and spectroscopic methods. The molecular masses, the pIs, and the UV-vis absorption spectra of the ferric and oxidized states of these LiP variant enzymes were similar to those of wild-type LiP (wtLiP), suggesting the overall protein and heme environments were not significantly affected by these mutations. The steady-state and transient-state parameters for the oxidation of veratryl alcohol (VA) by the H82A and Q222A variants were very similar to those of wtLiP, demonstrating that these residues are not involved in VA oxidation and that the heme access channel is an unlikely site for VA oxidation. In contrast, the W171A variant was unable to oxidize VA, confirming the apparent essentiality of Trp171 in VA oxidation by LiP. The kinetic rates of spontaneous LiP compound I reduction in the absence of VA were similar for W171A and wild-type LiP, suggesting that there may not be a radical formed on the Trp171 residue of LiP in the absence of VA. For the F267L variant, both the K(m app) value in the steady state and the apparent dissociation constant (K(D)) for compound II reduction were greater than those for wtLiP. These results indicate that the site including W171 and F267, rather than the heme access channel, is the site of VA binding and oxidation in LiP. Whereas Trp171 appears to be essential for VA oxidation, it apparently is not independently responsible for the spontaneous decomposition of oxidized intermediates. The nearby Phe267 apparently is also involved in VA binding. PMID- 11876659 TI - Initial disulfide formation steps in the folding of an omega-conotoxin. AB - To determine whether the native disulfides of omega-conotoxins are preferentially stabilized early in the folding of these small proteins, the rates and equilibria for disulfide formation were measured for three analogues of omega-conotoxin MVIIA. In each analogue, one of the three pairs of disulfide-bonded Cys residues was replaced with Ala residues, leaving four Cys residues that can form six intermediates with one disulfide and three species with two disulfides. For each analogue, all of the disulfide-bonded species were identified, and the equilibrium constants for forming the individual species via exchange with oxidized and reduced glutathione were measured. These equilibrium constants represent effective concentrations of the Cys thiols and ranged from 0.01 to 0.4 M in the fully reduced protein. There was little or no preference for forming the native disulfides, and the equilibria for forming the first and second disulfides decreased only slightly upon the addition of 8 M urea. The data for the four-Cys analogues, together with equilibrium data for the six-Cys form, were also used to estimate effective concentrations for forming a third disulfide once two native disulfides are present. These effective concentrations were approximately 100 and 10 M in the presence of 0 and 8 M urea, respectively. The results indicate that there is little or no preferential formation of native interactions in the folding of these molecules until two disulfides have formed, after which there is a high degree of cooperativity among the native interactions. PMID- 11876660 TI - Quinolinate phosphoribosyltransferase: kinetic mechanism for a type II PRTase. AB - Quinolinate phosphoribosyltransferase (QAPRTase, EC 2.4.2.19) catalyzes the formation of nicotinate mononucleotide, carbon dioxide, and pyrophosphate from 5 phosphoribosyl 1-pyrophosphate (PRPP) and quinolinic acid (QA, pyridine 2,3 dicarboxylic acid). The enzyme is the only type II PRTase whose X-ray structure is known. Here we determined the kinetic mechanism of the enzyme from Salmonella typhimurium. Equilibrium binding studies show that PRPP and QA each form binary complexes with the enzyme, with K(D) values (53 and 21 microM, respectively) similar to their K(M) values (30 and 25 microM, respectively). Although neither PP(i) nor NAMN products bound well to the enzyme, 130-fold tighter binding of PP(i) (K(D) = 75 microM) and NAMN (K(D) = 6 microM) in a ternary complex was observed. Phthalic acid (K(D) = 21 microM) and PRPP each caused a 2.5-fold tightening of the other's binding. Isotope trapping experiments indicated that the E.QA complex is catalytically competent, whereas the E.PRPP complex could not be trapped. Pre-steady-state kinetics gave a linear rate of NAMN formation, indicating that on-enzyme phosphoribosyl transfer chemistry is rate-determining. Isotope trapping from the steady state revealed that nearly all QA and about one third of PRPP in ternary enzyme.QA.PRPP complexes could be trapped as the product. Substrate inhibition by PRPP was observed. These data demonstrate a predominantly ordered kinetic mechanism in which productive binding of quinolinic acid precedes that of PRPP. An E.PRPP complex exists as a nonproductive side branch. PMID- 11876661 TI - Treating children exposed to disasters. PMID- 11876662 TI - Attention-deficit/hyperactivity disorder: is it common? Is it overtreated? PMID- 11876663 TI - Psychosocial intervention for postdisaster trauma symptoms in elementary school children: a controlled community field study. AB - CONTEXT: Natural disasters negatively affect children's emotional and behavioral adjustment. Although treatments to reduce psychological morbidity following disasters are needed, it has been difficult to conduct treatment research in postdisaster environments because of the sensitivity of victims to perceived intrusiveness and exploitation. OBJECTIVE: To evaluate the efficacy of a public health--inspired intervention combining school-based screening and psychosocial treatment to identify and treat children with persistent disaster-related trauma symptoms. DESIGN: To identify children with continued high levels of trauma related symptoms 2 years after a major disaster, we conducted a community-wide school-based screening of disaster-exposed public elementary school children. Children with the highest levels of trauma-related symptoms were randomly assigned to 1 of 3 consecutively treated cohorts. Children in the cohorts awaiting treatment served as wait-list controls. Within each cohort, children were randomly assigned to either individual or group treatment to allow comparison of the efficacy of the 2 treatment modalities. SETTING: All 10 public elementary schools on the island of Kauai (one of the Hawaiian Islands) 2 years after Hurricane Iniki. PARTICIPANTS: All 4258 children in second through sixth grade were screened. The 248 children with the highest levels of psychological trauma symptoms were selected for treatment. INTERVENTION: Children were randomly assigned to either individual or group treatment provided by specially trained school-based counselors. Treatment comprised 4 sessions. MAIN OUTCOME MEASURES: The Kauai Reaction Inventory, a self-report measure of trauma symptoms, and the Child Reaction Inventory, a semistructured clinical interview for posttraumatic stress disorder symptoms. RESULTS: After treatment, children reported significant reductions in self-reported trauma-related symptoms. This symptom reduction was maintained at the 1-year follow-up. Clinical interviews also indicated that treated children had fewer trauma symptoms compared with untreated children. CONCLUSIONS: School-based community-wide screening followed by psychosocial intervention seems to effectively identify and reduce children's disaster-related trauma symptoms and may facilitate psychological recovery. While group and individual treatments did not differ in efficacy, fewer children dropped out of the group treatment. This approach may be applicable to screening and treating children exposed to a variety of large-scale disasters. PMID- 11876664 TI - How common is attention-deficit/hyperactivity disorder? Incidence in a population based birth cohort in Rochester, Minn. AB - CONTEXT: The frequency of occurrence of attention-deficit/hyperactivity disorder (AD/HD) is in dispute. This uncertainty has contributed to the concern that too many children in the United States are being treated with stimulant medication. OBJECTIVES: To determine the cumulative incidence of AD/HD in a population-based birth cohort and to estimate the prevalence of pharmacologic treatment for children who fulfill research criteria for AD/HD. DESIGN: Population-based birth cohort study. SETTING AND SUBJECTS: All children born between 1976 and 1982 in Rochester, Minn, who remained in the community after age 5 years (N = 5718). MAIN OUTCOME MEASURES: Medical and school records were reviewed for clinical diagnoses of AD/HD and supporting documentation (symptoms consistent with Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition criteria and positive results for AD/HD-related questionnaires). Research-identified cases were defined as: (1) "definite" AD/HD (clinical diagnosis and at least one type of supporting documentation); (2) "probable" AD/HD (clinical diagnosis but no supporting documentation or no clinical diagnosis but both types of supporting documentation); (3) "questionable" AD/HD (no clinical diagnosis, but at least one type of supporting documentation); and (4) "not AD/HD" (all other subjects). Information about pharmacologic treatment for AD/HD was abstracted for all subjects. RESULTS: The highest estimate of the cumulative incidence at age 19 years (with 95% confidence interval) of AD/HD (definite plus probable plus questionable AD/HD) was 16.0% (14.7-17.3). The lowest estimate (definite AD/HD only) was 7.4% (6.5-8.4). Prevalence of treatment with stimulant medication was 86.5% for definite AD/HD, 40.0% for probable AD/HD, 6.6% for questionable AD/HD, and 0.2% for not AD/HD. CONCLUSIONS: These results provide insight into the apparent discrepancies in estimates of the occurrence of AD/HD, with less stringent criteria resulting in higher cumulative incidence. Children who met the most stringent criteria for AD/HD were most likely to receive pharmacologic treatment. PMID- 11876665 TI - Two educational interventions to improve pediatricians' knowledge and skills in performing ankle and knee physical examinations. AB - BACKGROUND: Methods are needed to improve pediatricians' skills for physical examination of the ankle and knee. OBJECTIVE: To compare the effect of 2 methods of teaching the physical examination of the ankle and knee on the knowledge and skills of pediatricians. DESIGN: Prospective intervention trial with preintervention and postintervention tests. SETTING: Pediatricians' offices. PARTICIPANTS: Twenty-three practice groups of community pediatricians, including 75 pediatricians (74% of the eligible sample). INTERVENTIONS: Practice groups were assigned by clustered randomization to 1 of 2 teaching interventions: (1) a videotape showing correct performance of the ankle and knee physical examinations (videotape only; 15 groups) and (2) the same videotape plus a skills building session (18 groups). The randomization was stratified by practice size. MAIN OUTCOME MEASURES: Knowledge of ankle and knee examinations and ankle and knee physical examination skills, assessed by means of a Clinical Skills Assessment Examination (CSAE), at 18 weeks. RESULTS: The pediatricians' baseline mean ankle CSAE score was 26% correct in both groups. These scores improved to 44% and 59% correct in the videotape and videotape plus skills instruction groups, respectively, at 18 weeks (P<.001 for both). The baseline mean knee CSAE score was 25% in both groups and improved to 30% (P =.02, videotape group) and 41% (P<.001, videotape plus skills instruction group) at 18 weeks. The change in CSAE scores was greater in the videotape plus skills instruction group. Written test scores improved significantly in both groups. CONCLUSIONS: Pediatricians' skills were lacking at baseline. Both teaching interventions were associated with improved skills and knowledge. PMID- 11876666 TI - Effect of growth hormone therapy on height in children with idiopathic short stature: a meta-analysis. AB - CONTEXT: Use of growth hormone (GH) therapy to promote growth in children with idiopathic short stature is controversial. A fundamental issue underlying the controversy is uncertainty about the magnitude of effectiveness of GH for this condition. OBJECTIVE: To determine the effect of GH on short- and long-term growth in idiopathic short stature. STUDY DESIGN: Systematic review of controlled and uncontrolled studies. DATA SOURCES: MEDLINE (1985-2000), key journals, cross referencing of bibliographies, abstract booklets, and experts. STUDY SELECTION AND DATA EXTRACTION: We performed a meta-analysis of all studies satisfying the inclusion criteria for idiopathic short stature: initial height below the 10th percentile, normal stimulated GH levels (>10 microg/L), absence of comorbid conditions, no previous GH therapy, treatment with biosynthetic GH, and inclusion of major outcome measures. PRIMARY OUTCOME MEASURES: Growth velocity and height SD score (number of SDs from mean height for age and sex) at baseline and after 1 year to evaluate the short-term effect of GH. Adult height was analyzed to evaluate the long-term effect of GH. DATA SYNTHESIS: Ten controlled trials (434 patients) and 28 uncontrolled trials (655 patients) met the inclusion criteria. While baseline growth velocities were equivalent at baseline, 1-year growth velocity of the GH-treated group significantly exceeded that of controls by 2.86 cm/y. Similarly, in uncontrolled trials, growth velocity increased after 1 year, and height SD score increased from -2.72 at baseline to -2.19. In controlled studies, the adult height of the GH-treated group significantly exceeded controls by 0.84 SD, and in uncontrolled trials the adult height attained after GH treatment (-1.62 SDs) exceeded that predicted at baseline (-2.18 SDs). These results suggest an average gain in adult height of approximately 4 to 6 cm (range, 2.3-8.7 cm) with GH therapy. Given current treatment costs, this corresponds to more than $35 000 per inch (2.54 cm) gained in adult height in idiopathic short stature. CONCLUSIONS: Treatment with GH results in short-term increases in growth for children with idiopathic short stature, and long-term GH can increase adult height. These results are fundamental to decisions about GH use and raise questions about the goals of treatment. Use of GH for idiopathic short stature in clinical practice will depend on its efficacy in promoting growth and the value of this effect to families, physicians, and third-party payers. PMID- 11876667 TI - Day care attendance, respiratory tract illnesses, wheezing, asthma, and total serum IgE level in early childhood. AB - BACKGROUND: It has been hypothesized that day care--related infections may explain the inverse relation between day care attendance in early life and asthma in childhood. OBJECTIVE: To examine the relation between day care attendance or respiratory tract illnesses in the first year of life and wheezing and asthma in the first 4 years of life among children with a parental history of atopy who were followed up from birth. RESULTS: Day care attendance in the first year of life was inversely associated with geometric mean total serum IgE level (12.9 [+/ 1 SD = 3.3, 51.4] IU/mL for day care vs 18.5 [[+/-1 SD = 5.3, 64.7] IU/mL for no day care; P =.03) at 2 years of age but not significantly associated with wheezing at or after 2 years of age. Having at least 1 physician-diagnosed lower respiratory tract illness in the first year of life was significantly associated with recurrent wheezing (odds ratio [OR], 2.0; 95% confidence interval [CI], 1.0 4.1) and asthma (OR, 2.5; 95% CI, 1.1-5.5) at 4 years of age, but not with any wheezing (infrequent and frequent) at 3 years or older. Illnesses of the upper respiratory tract (> or = 1 physician-diagnosed upper respiratory tract illness or > or = 3 episodes of nasal catarrh) in the first year of life were associated with any wheezing (frequent and infrequent) between the ages of 1 and 4 years, but not with recurrent wheezing or asthma at 4 years of age. CONCLUSIONS: Our results suggest that among children with a parental history of atopy the protective effect of day care attendance in early life against the development of atopy has begun by 2 years of age, and that a protective effect of day care attendance in early life against wheezing may not be observed until after 4 years of age. PMID- 11876668 TI - Health care use by children diagnosed as having developmental delay. AB - BACKGROUND: Although children with developmental delay are known to have increased health care use, it is unclear what proportion of that health care use is related to associated chronic health conditions. OBJECTIVES: To assess the prevalence of isolated developmental delay and to determine the role of developmental delay in health care use controlling for chronic health conditions. DESIGN: Retrospective cohort study using Washington State Medicaid claims records from November 1, 1990, to December 31, 1997, an administrative data set that contains both International Classification of Diseases, Ninth Revision, Clinical Modification codes and billed services. PATIENTS AND SETTING: Children born between November 1, 1990, and December 31, 1992, diagnosed as having developmental delay before the age of 5 years, enrolled in Medicaid within 1 month of birth, and continuously enrolled for at least 12 months. Four control subjects per case were matched on date of birth and duration of continuous enrollment in Medicaid. MAIN OUTCOME MEASURES: Visits to physicians, emergency departments, other practitioners, or hospitals by year of life. RESULTS: One thousand two hundred forty-two children having developmental delay and 5370 children without developmental delay were included. One percent of those who met study criteria had developmental delay without chronic health conditions and 30% of the children with developmental delay had no associated chronic health conditions. Boys were 1.6 times as likely to have a diagnosis of developmental delay. Developmental delay was independently associated with increased health care use by all 4 measures used. CONCLUSION: Developmental delay increases health care use apart from associated chronic health conditions. PMID- 11876669 TI - Intracranial hemorrhage in children younger than 3 years: prediction of intent. AB - OBJECTIVE: To determine whether certain computed tomographic imaging patterns in infants and young children with intracranial hemorrhage help predict intentional compared with unintentional injuries. DESIGN: Retrospective consecutive case series over a 10-year period. PATIENTS: Two hundred ninety-three children younger than 3 years with intracranial hemorrhage. MAIN OUTCOME MEASURES: The sensitivity and specificity of computed tomographic imaging patterns for intentional head injury. SETTING: Regional pediatric medical center. RESULTS: Four variables used in the multiple logistic regression analysis for predicting intentional head injury were statistically significant (P<.05): subdural hematoma located over the cerebral convexities, hematoma within the interhemispheric subdural space, hygroma (nonhemic subdural fluid) with intracranial hemorrhage, and absence of a skull fracture with intracranial hemorrhage. The prediction model for the diagnosis of intentional head trauma using combinations of these 4 variables and a.45 probability cutoff point indicated a sensitivity of 84% (95% confidence interval, 78%-90%) and a specificity of 83% (95% confidence interval, 74%-89%). CONCLUSION: Computed tomographic imaging patterns of intracranial hemorrhage in children younger than 3 years help predict whether the injury was intentional. PMID- 11876670 TI - Parent-reported environmental exposures and environmental control measures for children with asthma. AB - BACKGROUND: Mounting evidence suggests that indoor allergens and irritants contribute to childhood asthma. National asthma guidelines highlight the importance of their reduction as part of comprehensive asthma treatment. OBJECTIVES: To assess the prevalence of potential environmental triggers, to identify risk factors for such exposures, and to determine whether prior parental education about trigger avoidance is associated with fewer such exposures. SETTING AND PATIENTS: Children with asthma in practices affiliated with 3 managed care organizations. INTERVENTIONS: Parents of 638 children, aged 3 to 15 years, were interviewed on enrollment in a randomized trial of asthma care improvement strategies. Parents reported recent asthma symptoms and exposures to potential environmental triggers. Multivariate models were used to identify specific demographic risk factors for environmental exposures and to determine if prior education was associated with fewer such exposures. RESULTS: Exposures to environmental triggers were frequent: 30% of households had a smoker, 18% had household pests, and 59% had furry pets. Other exposures included bedroom carpeting (78%) and forced-air heat (58%). Most children did not have mattress (65%) or pillow (84%) covers. Of the parents, 45% reported ever receiving written instructions regarding trigger avoidance and 11% reported them given in the past year. However, 42% reported discussing triggers in the home environment with a clinician in the past 6 months. In multivariate models, predictors of smoking at home included low annual family income and lower parental educational attainment. Dog ownership was associated with low educational attainment, and dog and cat ownership were less likely with black race. Reports of pests were increased for black children compared with white children. Black race was associated with lower rates of other exposures, including bedroom carpeting. After controlling for potential confounders, there was no association of reduced exposures with prior receipt of environmental control instructions. CONCLUSIONS: Exposure to potential environmental triggers is common, and recommended trigger avoidance measures are infrequently adopted. While specific exposures may vary with demographic and socioeconomic variables, all children are at risk. New methods for educating parents to reduce such exposures should be tested. PMID- 11876671 TI - Experience with wood lamp illumination and digital photography in the documentation of bruises on human skin. AB - Bruising is very common in children. Examination of bruising can guide the clinician in ordering radiographic imaging studies of children who have suffered trauma. Additionally, bruising in infants and patterns of bruising that do not match the injury scenario offered by caretakers can raise the suspicion of abuse. This article reports preliminary experience with Wood lamp enhancement of faint bruises and visualization of bruises that are not visible. It describes the method for digital photography of bruises visualized in this way. Finally, it suggests future applications and areas of further study. PMID- 11876672 TI - Identification of population subgroups of children and adolescents with high asthma prevalence: findings from the Third National Health and Nutrition Examination Survey. AB - OBJECTIVES: To provide national estimates of asthma prevalence in African American, Mexican American and white (non-Latino) children and adolescents using several common definitions; to evaluate familial, sociodemographic, and environmental risk factors that are independently associated with current asthma in children; and to identify subgroups at particular risk for current asthma using 2 complementary data analytic approaches. DESIGN: Cross-sectional study, using the Third National Health and Nutrition Examination Survey, 1988-1994. SETTING: Eighty-nine mobile examination centers in the United States. PARTICIPANTS: Twelve thousand three hundred eighty-eight African American, Mexican American, and white (non-Latino) children and adolescents, aged 2 months through 16 years, selected from a systematic random, population-based, nationally representative sample. MAIN OUTCOME MEASURE: Current asthma, defined by caregivers who reported that their child currently had doctor-diagnosed asthma. RESULTS: The overall prevalence of current asthma was 6.7% (95% confidence interval [CI], 5.6-7.8). Odds ratios for current asthma from the multiple regression analysis were 4.00 (95% CI, 2.90-5.52) for children with a parental history of asthma or hay fever, 1.94 (95% CI, 1.09-3.46) for children with body mass index (calculated as weight in kilograms divided by the square of height in meters) greater than or equal to the 85th percentile, and 1.64 (95% CI, 1.20 2.26) for children of African American ethnicity. African American and Mexican American children showed a consistent prevalence of current asthma across age while white children showed an increase in prevalence with age. The 2 highest risk subgroups identified by the signal detection analysis were composed of children with a parental history of asthma or hay fever who were 10 years or older with a body mass index greater than or equal to the 85th percentile (31.0% current asthma), and children with a parental history who were 10 years or younger and of African American ethnicity (15.6% current asthma). CONCLUSIONS: The findings from this analysis show a strong independent association between obesity and current asthma in children and adolescents, and confirm previous reports of a parental history of asthma or hay fever and African American ethnicity as additional important risk factors. PMID- 11876673 TI - Neonatal jaundice in Asian, white, and mixed-race infants. AB - BACKGROUND: East Asians have inherently higher bilirubin levels at birth than whites. The potential for unnecessary treatment makes jaundice a problem of public health and clinical significance. OBJECTIVES: To report the occurrence of jaundice diagnoses in East Asian and mixed East Asian/white infants in Washington State in recent years, and to compare the risk of diagnosis with neonatal jaundice among these infants, relative to white infants. DESIGN: Population-based cohort study in Washington state. Participants were infants of full East Asian parentage (n = 3000), maternal Asian parentage (n = 2997), paternal Asian parentage (n = 2048), and white parentage (n = 3000). Diagnoses of jaundice and "severe jaundice" were identified using International Classification of Diseases, Ninth Revision (ICD-9) diagnosis and procedure codes from hospital discharge records. RESULTS: Infants of full East Asian parentage were more likely to be diagnosed with jaundice than were white infants (relative risk [RR], 1.37; 95% confidence interval [CI], 1.16-1.62). For infants with Asian mothers and white fathers, the RR was 1.09 (95% CI, 0.91-1.30). Infants with Asian fathers and white mothers had an RR of 1.26 (95% CI, 1.05-1.52). The risk of severe jaundice requiring phototherapy, blood transfusion, or rehospitalization, however, was significantly elevated only for infants of full East Asian parentage (RR, 1.7; 95% CI, 1.12-2.58). CONCLUSIONS: Diagnoses of neonatal jaundice occurred more often among East Asian and mixed Asian/white infants than among white infants. However, the risk of jaundice requiring extended hospital stay, rehospitalization, phototherapy, or blood transfusion was elevated only for infants of full East Asian parentage. PMID- 11876674 TI - Violence exposure, trauma, and IQ and/or reading deficits among urban children. AB - BACKGROUND: Exposure to violence in childhood has been associated with lower school grades. However, the association between violence exposure and performance on standardized tests (such as IQ or academic achievement) in children is unknown. It is also not known whether violence exposure itself or subsequent symptoms of trauma are primarily responsible for negative outcomes. OBJECTIVE: To examine the relationship between violence exposure and trauma-related distress and standardized test performance among early school-aged urban children, controlling for important potential confounders. DESIGN: A total of 299 urban first-grade children and their caregivers were evaluated using self-report, interview, and standardized tests. MAIN OUTCOME MEASURES: The child's IQ (Wechsler Preschool and Primary Scale of Intelligence--Revised) and reading ability (Test of Early Reading Ability, second edition) were the outcomes of interest. RESULTS: After controlling for confounders (child's gender, caregiver's IQ, home environment, socioeconomic status, and prenatal exposure to substance abuse) violence exposure was related to the child's IQ (P =.01) and reading ability (P =.045). Trauma-related distress accounted for additional variance in reading ability (P =.01). Using the derived regression equation to estimate effect sizes, a child experiencing both violence exposure and trauma-related distress at or above the 90th percentile would be expected to have a 7.5-point (SD, 0.5) decrement in IQ and a 9.8-point (SD, 0.66) decrement in reading achievement. CONCLUSION: In this study, exposure to violence and trauma-related distress in young children were associated with substantial decrements in IQ and reading achievement. PMID- 11876675 TI - Maternal ratings of infant intensity and distractibility: relationship with crying duration in the second month of life. AB - OBJECTIVE: To investigate the relationship between infant temperament characteristics assessed at 4 weeks of age and the duration of infant crying and fussing during the second month of life. DESIGN: Families were enrolled in this prospective study during prenatal classes, and 60 infants completed the study. Temperament was assessed when the infant was 4 weeks of age using the Early Infancy Temperament Questionnaire, and crying and fussing was assessed on 16 days during the second month of life using a parent-completed infant behavior diary. RESULTS: Ratings of the total duration of infant crying and fussing correlated significantly with the sum score on the temperament questionnaire (r = 0.36; P =.005). A longer duration of crying and fussing was associated with infants with high intensity (r = 0.43; P =.001) and low distractibility (r = 0.37; P =.003). CONCLUSIONS: The finding that mothers rating their infants as having high intensity and low distractibility is associated with increased crying duration supports a growing body of literature suggesting that infants with high levels of crying are more reactive to sensory stimuli and harder to soothe than those who cry less. Physicians counseling parents of infants with persistent crying should recognize the infant characteristics associated with increased crying. PMID- 11876677 TI - Radiological case of the month. Intracranial hypertension and reduced cerebral blood flow in meningococcal meningitis. PMID- 11876676 TI - Pathological case of the month. Celiac disease with acanthocytosis. PMID- 11876678 TI - The pediatric forum: ecstasy use among club rave attendees. PMID- 11876679 TI - Some pitfalls of computer modeling. PMID- 11876681 TI - Medical neglect: a child-focused view. PMID- 11876685 TI - Review article: the non-inherited gastrointestinal polyposis syndromes. AB - The non-inherited gastrointestinal polyposis syndromes represent a group of rare disorders characterized by the presence of multiple, non-adenomatous polyps on the gastrointestinal mucosa occurring in unrelated patients. We present here a review of the clinical and histo- pathological aspects of the syndromes to include the Cronkhite-Canada syndrome, hyperplastic polyposis and lipomatous polyposis. While infrequently encountered, these diseases can have devastating clinical effects that may be aggravated by delays in diagnosis and treatment. Prompt accurate diagnosis and treatment of these uncommon disorders depend on a sound working knowledge of the distinct clinical and pathological features described herein. PMID- 11876686 TI - Review article: the complexity of drug development for irritable bowel syndrome. AB - Drug development for functional gastrointestinal disorders is complex. These conditions involve central and peripheral physiological changes, together with psychological factors. Methodological problems have included a poor appreciation of the physiological and psychological correlates of patients' symptoms, a lack of animal models of proven relevance, and safety issues. Government, patient pressure groups and the Internet can also influence a drug's success. Most recent interest has focused on the serotonin (5-HT) modifying drugs. Cisapride has been withdrawn in some countries because of concerns related to QT prolongation and cardiac arrhythmias. The 5-HT3 antagonists, developed to modify visceral sensation, have caused constipation; alosetron, also withdrawn, caused ischaemic colitis. The 5-HT4 agonists induce peristalsis; tegaserod and prucalopride, both delayed in their development due to issues of safety and efficacy, benefit patients with 'constipation-predominant' irritable bowel syndrome or idiopathic constipation. 5-HT1 agonists improve impaired gastric accommodation and symptoms in patients with functional dyspepsia. Antidepressants also affect serotonin metabolism. Previous examples of success in this area involved drugs targeted at peripheral receptors mediating motor function or secretion. Modification of sensory function is a much more challenging objective. The experience with serotonin modifying drugs has been mixed, and some important lessons are there to be learnt. PMID- 11876687 TI - Chemotherapy for colon cancer in a well-defined French population: is it under- or over-prescribed? AB - BACKGROUND: It has been demonstrated that adjuvant chemotherapy in TNM stage III and palliative chemotherapy are effective treatments for colon cancer. AIM: To determine changes over a 10-year period in the practice of adjuvant and palliative chemotherapy for colon cancer in a well-defined French population. METHODS: Some 4093 patients with colon adenocarcinoma diagnosed between 1989 and 1998 were studied. To estimate the independent effect of the studied variables, a non-conditional logistical regression was performed. RESULTS: The proportion of patients with stage II disease treated with adjuvant chemotherapy increased from 2.3% (1989-90) to 20.5% (1997-98). The corresponding figures for stage III patients were 7.1% and 54.9%. This increase was particularly marked in younger patients, with 47.3% of stage II and 86.1% of stage III patients under the age of 65 years being treated in the 1997-98 period, compared with 4.9% and 24.4% of those over 75 years for the same period (P < 0.0001). The use of palliative chemotherapy increased over time from 13.6% (1989-90) to 38.9% (1997-98). Again, this increase was more dramatic in the younger age group. CONCLUSIONS: The use of adjuvant chemotherapy has increased for stage II disease despite the absence of proven effectiveness. Both adjuvant and palliative chemotherapy are still under prescribed in patients over the age of 75 years. PMID- 11876688 TI - Flumazenil vs. placebo in hepatic encephalopathy in patients with cirrhosis: a meta-analysis. AB - BACKGROUND: Randomized controlled trials testing flumazenil in hepatic encephalopathy have shown conflicting results. AIM: To compare flumazenil and placebo in hepatic encephalopathy in patients with cirrhosis. METHODS: An overview of randomized controlled trials comparing flumazenil and placebo in hepatic encephalopathy in patients with cirrhosis was performed. For each end point, heterogeneity and treatment efficacy were assessed by Peto and Der Simonian methods. As most trials were crossover in nature, a sensitivity analysis was performed including the two treatment periods. RESULTS: Six double-blind randomized controlled trials, including 641 patients (326 treated with flumazenil and 315 with placebo), were identified. The treatment duration ranged from 5 min to 3 days. Heterogeneity tests between control groups were not significant. The mean percentages of patients with clinical improvement (five trials) were 27% in treated groups and 3% in placebo groups. This difference was significant by both methods (Peto: odds ratio=6.15; 95% confidence interval, 4.0-9.5; P < 0.001; Der Simonian: mean rate difference, 29%; 95% confidence interval, 17-41; P < 0.001). The mean percentages of patients with electroencephalographic improvement were 19% in treated groups and 2% in placebo groups. This difference was significant only with the Peto method (odds ratio=5.8; 95% confidence interval, 3.4-9.7; P < 0.001). The sensitivity analysis showed similar results. CONCLUSIONS: This meta analysis shows that flumazenil induces clinical and electroencephalographic improvement of hepatic encephalopathy in patients with cirrhosis. PMID- 11876689 TI - Haemodynamic effects of acute and chronic administration of low-dose carvedilol, a vasodilating beta-blocker, in patients with cirrhosis and portal hypertension. AB - BACKGROUND: Carvedilol is a non-selective vasodilating beta-blocker with weak alpha1 receptor antagonism. Recent studies have demonstrated its potential as a portal hypotensive agent. AIM: To assess the haemodynamic effects and patient tolerability of the acute and chronic administration of low-dose carvedilol. METHODS: Haemodynamic measurements were performed in ten cirrhotic patients before and 1 h after the administration of 12.5 mg oral carvedilol. The study was repeated 4 weeks after daily administration of 12.5 mg carvedilol. RESULTS: After acute administration of carvedilol, there was a 23% reduction in the hepatic venous pressure gradient from 16.37 +/- 2.14 to 12.56 +/- 3.91 mmHg (P < 0.05), with significant falls in the heart rate, mean arterial pressure and cardiac output. Chronic administration resulted in a further fall in the hepatic venous pressure gradient from a baseline of 16.37 +/- 0.71 to 9.27 +/- 1.40 mmHg (P < 0.001) with the mean arterial pressure being unaffected. The drug was well tolerated with only one patient experiencing asymptomatic hypotension. CONCLUSIONS: The results show that low-dose carvedilol is an extremely potent portal hypotensive pharmacological agent, and is worthy of further investigation in large randomized trials to assess its effect in preventing variceal haemorrhage. PMID- 11876690 TI - A randomized trial of high-dose interferon alpha-2b, with or without ribavirin, in chronic hepatitis C patients who have not responded to standard dose interferon. AB - BACKGROUND: Conventional interferon monotherapy fails to achieve virological clearance in most hepatitis C-infected patients. The use of high-dose induction regimens may improve the initial clearance of virus, while the addition of ribavirin appears to improve the rates of sustained response once clearance is achieved. AIM: To compare the efficacy and safety of re-treatment with an induction regimen of high-dose interferon alpha-2b, with or without ribavirin, in chronic hepatitis C patients who have not responded to standard dose interferon monotherapy. METHODS: Previous virological non-responders to standard dose interferon (3-5 MU three times weekly for > or = 12 weeks) were randomized to receive, unblind, either 10 MU interferon alpha-2b daily for 10 days, then 5 MU daily for 74 days, then 5 MU three times weekly for 24 weeks (total 36 weeks) (group A), or the above regimen with the addition of ribavirin, 1000-1200 mg/day, at day 11 (group B). All patients were followed up for 24 weeks after completion of therapy. RESULTS: End of treatment virological response was noted in one of 10 (10%) patients in group A and in eight of 15 (54%) patients in group B (P=0.04). The sole end treatment responder in group A and three in group B relapsed on follow-up. The apparent improvement in response in group B compared to group A nearly reached statistical significance (group B 5/15 vs. group A 0/10; P=0.06). CONCLUSIONS: In this small pilot study, a 36-week high-dose induction interferon monotherapy protocol did not yield sustained responses in previous non-responders to standard dose interferon. However, the same regimen with ribavirin yielded a 33% sustained response rate, nearly reaching statistical significance. The therapy was well tolerated, despite the higher doses of interferon used and the addition of ribavirin. High-dose interferon with ribavirin appears to be a therapeutic option for non-responders to conventional interferon monotherapy. PMID- 11876691 TI - Relevance of thiopurine methyltransferase activity in inflammatory bowel disease patients maintained on low-dose azathioprine. AB - BACKGROUND: It is well-recognized that patients with low thiopurine methyltransferase activity are more susceptible to the development of bone marrow suppression side-effects. AIM: To study the impact of thiopurine methyltransferase activity on the clinical course of inflammatory bowel disease patients treated with low-dose azathioprine (< 2 mg/kg). METHODS: We measured the thiopurine methyltransferase activity of blood samples from 113 inflammatory bowel disease patients who were taking azathiopurine, had discontinued azathioprine because of side-effects, or had never taken azathioprine. The thiopurine methyltransferase activity was compared with that of 17 healthy controls. Relapse rates and time to first relapse were compared in inflammatory bowel disease patients and stratified according to their thiopurine methyltransferase activity. RESULTS: Patients who became neutropenic had a significantly lower mean thiopurine methyltransferase activity than that of patients who developed other side-effects (analysis of variance, P < 0.05). Survival curves were constructed (time to first relapse) for patients treated with low-dose azathioprine for thiopurine methyltransferase activities of < 20 and > 20 nmol/mL red blood cells/h. There was a significantly lower number of relapses in inflammatory bowel disease patients with lower thiopurine methyltransferase levels (P < 0.05). CONCLUSIONS: The mean thiopurine methyltransferase activity was significantly lower in patients on a low dose of azathioprine in remission compared with those who relapsed. The thiopurine methyltransferase activity was significantly lower in patients who discontinued azathioprine due to neutropenia than in those who discontinued due to other side effects. PMID- 11876692 TI - Randomized, controlled trial of recombinant human interleukin-11 in patients with active Crohn's disease. AB - BACKGROUND: Interleukin-11 is a mesenchymally derived cytokine with pleiotropic activities. A pilot study suggested therapeutic benefit of recombinant human interleukin-11 (rhIL-11) in patients with Crohn's disease. AIM: To determine the safety and preliminary estimate of efficacy of rhIL-11 in treating active Crohn's disease. METHODS: Patients with mild to moderately active Crohn's disease, defined as a Crohn's disease activity index (CDAI) > or = 220 and < or = 450, were enrolled in a multicentre trial. Stable doses of 5-aminosalicylates, antibiotics, 6-mercaptopurine or azathioprine were permitted with appropriate wash-in periods. Oral, intravenous or rectally administered corticosteroids were not allowed. Patients were randomized to 6 weeks of subcutaneous injection with rhIL-11 15 microg/kg or placebo weekly, or rhIL-11 7.5 microg/kg or placebo twice weekly. The primary end-point was per cent change in CDAI at week 6; the major secondary end-point was the proportion of patients in remission, defined as a 100 point decrease in CDAI and absolute CDAI < or = 150. RESULTS: Baseline characteristics were similar among the 148 evaluated patients (49 placebo, 49 rhIL-11 15 microg/kg once weekly, 50 rhIL-11 7.5 microg/kg twice weekly). Treatment was well-tolerated, with mild injection site reactions occurring more frequently among patients treated with rhIL-11. Headache, oedema, and increased platelet count occurred significantly more often in the rhIL-11 7.5 microg/kg twice weekly group, but not the 15 microg/kg once weekly group. There was a trend toward decreased mean per cent change in CDAI in the rhIL-11 15 micro/kg once weekly group vs. placebo (-31.5% vs. -18.5%, 95% confidence interval for the difference -27.9-1.6%). A significantly greater proportion of patients receiving rhIL-11 15 microg/kg once weekly achieved remission compared to placebo (36.7% vs. 16.3%, 95% confidence interval for the difference 3.4-37.4%; 16.4% for rhIL 11 7.5 microg/kg, N.S.). CONCLUSIONS: Weekly subcutaneous injection with rhIL-11 15 microg/kg is safe and effective in inducing remission in a subset of patients with active Crohn's disease. PMID- 11876693 TI - Treatment of mildly to moderately active ulcerative colitis with a tryptase inhibitor (APC 2059): an open-label pilot study. AB - BACKGROUND: Mast cells isolated from the colonic mucosa in active ulcerative colitis appear to be partially degranulated, suggesting the release of tryptase. AIM: To investigate the safety and activity of APC 2059, a highly specific tryptase inhibitor, in the treatment of ulcerative colitis. METHODS: This was an open-label, Phase 2, multicentre pilot study in patients with mildly to moderately active ulcerative colitis, with a disease activity index of 6-9 on a 12-point scale. Fifty-six adults received 20 mg APC 2059 subcutaneously twice daily and 53 completed 28 days of treatment. The primary end-point was response, defined as a final disease activity index of < or = 3. Supplementary analyses were also performed. RESULTS: Sixteen (29%) of 56 patients responded. Five (9%) showed complete remission (disease activity index=0). Twenty-seven (49%) improved, with a final disease activity index of < or = 3 or a four-point reduction. Improvement or normalization in each category of the disease activity index was as follows: stool frequency, 64%; bleeding, 64%; endoscopy, 50%; physicians' rating, 63%. There were no significant relationships between outcome and pharmacokinetics. The most common adverse events were related to the injection site (32.1%). CONCLUSIONS: In this pilot study, the tryptase inhibitor APC 2059 was safe and there was evidence of activity in the treatment of ulcerative colitis. PMID- 11876694 TI - A new mesalazine foam enema (Claversal Foam) compared with a standard liquid enema in patients with active distal ulcerative colitis. AB - BACKGROUND: Rectally administered mesalazine (5-aminosalicylic acid) is a recognized therapy for distal ulcerative colitis. It is frequently applied as a liquid enema. However, there are reasons (acceptability to the patient, more uniform topical dispersion and effective adhesion) to prefer a foam-based enema. AIM: This study compared a foam enema (2 g mesalazine per day, Claversal Foam) with a standard liquid enema (4 g mesalazine per day, Salofalk enema). METHODS: Patients with active distal ulcerative colitis, diagnosed according to standardized criteria, were treated for 4 weeks. The primary goal was clinical remission; endoscopic remission, histological changes, global assessment and standard safety measures were also analysed. A major subset of the patients also provided quality-of-life data. RESULTS: Both foam and liquid enema gave good rates of clinical and endoscopic remission. The foam enema was shown to be as efficacious as the reference, even though the daily dose in the foam treatment contained only half as much active drug as in the reference treatment. Minor regional differences in efficacy were seen. The tolerabilities of the two formulations were comparable. CONCLUSIONS: The foam enema offers a safe, efficacious and acceptable treatment for distal ulcerative colitis. PMID- 11876695 TI - Acid suppression in healthy subjects following lansoprazole or pantoprazole. AB - AIM: To compare the effect of lansoprazole, 30 mg once daily, with that of pantoprazole, 40 mg once daily, for the inhibition of gastric acid secretion. METHODS: Two randomized, single-blind, two-way, crossover studies were conducted in 74 healthy male volunteers. Lansoprazole, 30 mg, or pantoprazole, 40 mg, was administered once daily for five consecutive days with at least a 2-week washout period between regimens. Ambulatory 24-h intragastric pH was recorded at baseline and on days 1 and 5 of each crossover treatment period. RESULTS: On day 1 in both studies, lansoprazole, 30 mg, produced significantly higher mean 24-h intragastric pH values when compared to pantoprazole, 40 mg (3.78 vs. 3.08, P < 0.001, and 3.97 vs. 3.20, P < 0.001, in the first and second studies, respectively). In both studies, lansoprazole, 30 mg, produced significantly greater proportions of time that the intragastric pH was above 3, 4 and 5 when compared with pantoprazole, 40 mg (P < 0.005 in all comparisons). By treatment day 5 in the first study, lansoprazole, 30 mg, continued to produce a higher mean 24-h intragastric pH (4.15 vs. 3.91, P=0.014) and a significantly greater percentage of time that the intragastric pH was above 4 (63% vs. 56%, P=0.017) and 5 (41% vs. 30%, P < 0.001) when compared with pantoprazole, 40 mg. In the second study, the effects on intragastric pH were comparable between the two treatment groups. Headache was the most commonly reported adverse experience (nine lansoprazole-treated subjects, seven in the first study and two in the second study; six pantoprazole-treated subjects, five in the first study and one in the second study). CONCLUSIONS: Lansoprazole, 30 mg once daily, produces a faster onset and greater degree of acid inhibition than pantoprazole, 40 mg once daily. The implications for these differences on symptom relief and healing of erosive oesophagitis should be explored. PMID- 11876696 TI - Effects of antacid formulation on postprandial oesophageal acidity in patients with a history of episodic heartburn. AB - BACKGROUND: Heartburn self-treatment with antacids is extremely common. If the oesophagus is the primary site of antacid action, chewable antacids might raise the oesophageal pH more effectively than swallowable tablets. AIM: To establish a model to assess postprandial acid reflux and to compare the onset and duration of action on oesophageal pH of different antacid formulations. METHODS: Twenty subjects with a history of episodic heartburn underwent eight pH monitoring sessions each for 5.5 h postprandially. One hour after consuming a meal consisting of chili, cheese, raw onions and cola, subjects received 750 mg, 1500 mg and 3000 mg of either chewable or swallowable CaCO3 tablets, an effervescent bicarbonate solution or placebo. Oesophageal and gastric pH data were collected. RESULTS: Mean intra-oesophageal pH remained lower than baseline for more than 1 h (pH range 5-5.5) postprandially, indicating reflux of somewhat acidic intragastric contents into the oesophagus. The onset of action on oesophageal pH was similar for all antacids (30-35 min). The duration of action on pH varied: chewable tablets and effervescent bicarbonate had relatively long durations of action (oesophagus, 40-45 min; stomach, 100-180 min); swallowable tablets had little effect. CONCLUSIONS: The meal model used in this study dependably produced acidic gastro-oesophageal reflux. Antacids increased oesophageal pH independent of gastric pH, demonstrating that chewing antacids controls oesophageal acidity more effectively than swallowing antacid tablets. PMID- 11876697 TI - Onset of symptom relief with rabeprazole: a community-based, open-label assessment of patients with erosive oesophagitis. AB - BACKGROUND: In numerous clinical trials, proton pump inhibitors have demonstrated potent acid suppression and healing of erosive oesophagitis, as well as successful symptom relief for the entire spectrum of gastro-oesophageal reflux disease. AIM: The 'Future of Acid Suppression Therapy' (FAST) trial evaluated, in actual clinical practice, the timing of symptom relief, changes in symptom severity, health-related quality of life and safety in endoscopically confirmed erosive gastro-oesophageal reflux disease treated with rabeprazole. METHODS: This open-label, multicentre study enrolled 2579 patients to receive rabeprazole treatment using 20 mg once daily for 8 weeks. Between two clinical visits (at enrollment and week 8), patients used an interactive voice response system to rate gastro-oesophageal reflux disease symptoms. Subgroup analyses of efficacy were conducted for gender, age, Hetzel-Dent grade, presence of Barrett's oesophagus and for patients reporting previously ineffective symptom relief with omeprazole or lansoprazole. RESULTS: On day 1, rabeprazole significantly decreased daytime and night-time heartburn severity, regurgitation and belching. Complete relief of daytime and night-time heartburn was achieved in 64.0% and 69.2% of symptomatic patients, respectively, on day 1, and in 81.1% and 85.7% of patients, respectively, on day 7. Patients with moderate or severe heartburn symptoms at baseline achieved an even greater degree of satisfactory symptom relief (none or mild) from day 1 onwards. The median time to satisfactory heartburn relief was 2 days. Subgroup analyses showed no consistent differences in efficacy compared to the overall population treated. Health-related quality of life in patients was significantly lower than that of the US general population and improved significantly after 8 weeks of rabeprazole therapy. Rabeprazole was well tolerated, with headache as the most common adverse event, reported by less than 2% of the study population. CONCLUSIONS: In this large, open-label trial, rabeprazole rapidly and effectively relieved gastro-oesophageal reflux disease symptoms in most patients with erosive oesophagitis. Substantial symptom relief was noted on day 1; improvement continued over the first week and at week 4. By week 8, the health-related quality of life had also improved vs. baseline. PMID- 11876698 TI - Integrated acidity and rabeprazole pharmacology. AB - BACKGROUND: Integrated gastric and oesophageal acidity can be calculated from measurements of gastric and oesophageal pH and used to quantify gastric and oesophageal acidity over time. Rabeprazole is a new proton pump inhibitor that is effective in treating gastro-oesophageal reflux disease (GERD). AIM: To use measurement of integrated gastric and oesophageal acidity to determine the onset, duration and overall effect of rabeprazole in subjects with GERD. METHODS: Subjects with GERD were required to have oesophageal pH less-than-or-equal 4 for at least 10% of a 24-h recording. Effects of 20 mg rabeprazole on 24-h gastric and oesophageal pH were measured on days 1 and 7 of dosing. Integrated gastric and oesophageal acidity were calculated from time-weighted average hydrogen ion concentrations at each second of the 24-h record. RESULTS: At steady-state, 20 mg rabeprazole inhibited gastric acidity by 89% and oesophageal acidity by 95%. The first dose of rabeprazole inhibited gastric and oesophageal acidity by at least 70% of the steady-state effect. Oesophageal acidity could be divided into monophasic and biphasic patterns, and rabeprazole had different effects on oesophageal and gastric acidity in these two GERD subpopulations. The onset of action of the first dose of rabeprazole on gastric acidity was 4 h and on oesophageal acidity was 4 h in monophasic subjects and 7 h in biphasic subjects. Integrated acidity was more sensitive than time pH less-than-or-equal 4 in measuring the inhibitory actions of rabeprazole. CONCLUSIONS: Integrated gastric and oesophageal acidity are quantitative measurements that provide useful and novel information regarding the pathophysiology of GERD as well as the impact of antisecretory agents such as rabeprazole. PMID- 11876699 TI - Upper gastrointestinal evaluation of Chinese patients with non-cardiac chest pain. AB - AIMS: To test the usefulness of upper gastrointestinal investigations and quality of life assessment in Chinese patients with non-cardiac chest pain. METHODS: Seventy-eight consecutive patients with non-cardiac chest pain underwent upper endoscopy. Eight patients had upper gastrointestinal pathology (10%). The remaining 70 patients received acid perfusion test, oesophageal manometry and 24 h ambulatory oesophageal pH (n=65)/manometry (n=61), and the results were compared with those of healthy controls (n=20). Symptoms and quality of life (SF 36) were assessed by standard validated questionnaire. RESULTS: Significant acid reflux symptoms were present in five (5/70, 7%) patients. Abnormal 24-h oesophageal pH, indicating gastro-oesophageal reflux, was found in 19 (19/65, 29%) patients. The percentage of simultaneous contractions was higher and the percentage peristalsis was lower in patients with non-cardiac chest pain when compared with normal subjects by 24-h ambulatory manometry. Patients with non cardiac chest pain had a lower SF-36 score when compared to controls. CONCLUSIONS: Typical acid reflux symptoms are uncommon in Chinese patients with non-cardiac chest pain, but abnormal 24-h pH results, indicating gastro oesophageal reflux, were found in 29% of patients. Ineffective contractions were more frequently found in patients with non-cardiac chest pain by 24-h ambulatory manometry, which may have a bearing on the impaired quality of life in such patients. Upper gastrointestinal investigations are useful for the evaluation of Chinese patients with non-cardiac chest pain. PMID- 11876700 TI - Over-the-counter H(2)-receptor antagonists do not compromise intragastric pH control with proton pump inhibitors. AB - BACKGROUND: Proton pump inhibitors effectively suppress intragastric acid. Nocturnal acid breakthrough occurs on any dosing regimen of oral proton pump inhibitors. Histamine(2)-receptor antagonists (H(2)RA) suppress intragastric acidity independently of meals and help to control nocturnal acid breakthrough. Because proton pump inhibitors require an acid intragastric milieu for activation, nocturnal dosing of H(2)RA might decrease the effect of proton pump inhibitors taken in the morning by decreasing their gastric-acid-driven activation. AIM: Assess intragastric acid control on omeprazole, 20 mg, taken every morning, after variable dosing of over-the-counter famotidine, 10 mg. METHODS: Twelve Helicobacter pylori-negative, healthy volunteers received omeprazole, 20 mg, every morning before breakfast for 15 days. Baseline studies on omeprazole, 20 mg, in the morning, were done on day 7. On nights between days 8-9, 11-12 and 14-15, famotidine, 10 mg at bedtime, and 10 mg at bedtime and/or at 05.30 h, was given in a three-way, crossover, double-blind randomized design. Intragastric pH monitoring was performed on days 9, 12 and 15, starting at 08.00 h. RESULTS: Percentage times intragastric pH < 4 on omeprazole, 20 mg, in the morning of the day after receiving famotidine, 10 mg, at bedtime (58.6 +/- 4.8); at 05.30 h (54.1 +/- 5.1); or at bedtime and at 05.30 h (54.3 +/- 5.0) did not differ significantly (P=0.65) from percentage times intragastric pH on day 7 of omeprazole, 20 mg, in the morning (49.5 +/- 5.1). CONCLUSION: Concerns over inhibition of next-day daytime proton-pump inhibitor effect should not preclude use of nocturnal H(2)RAs in patients with gastro-oesophageal reflux disease. PMID- 11876701 TI - A randomized, double-blind, comparative study of standard-dose rabeprazole and high-dose omeprazole in gastro-oesophageal reflux disease. AB - BACKGROUND: Rabeprazole has a faster onset of antisecretory action than omeprazole, and it is of interest to determine whether this translates into faster symptom relief in patients with gastro-oesophageal reflux disease. AIMS: To assess the relief from heartburn after 3 days of treatment with standard-dose rabeprazole or high-dose omeprazole (primary end-point). Secondary end-points included the decrease in score for other symptoms of gastro-oesophageal reflux disease, healing rates and quantification of antacid use. METHODS: Patients with endoscopically confirmed erosive oesophagitis were randomized to receive 4 weeks of double-blind treatment with rabeprazole (20 mg) or omeprazole (40 mg). Patients who were not healed after 4 weeks received a further 4 weeks of treatment. RESULTS: Two hundred and seventy-four patients were screened, 251 patients were randomized and 230 patients completed the trial. The numbers of patients with relief from heartburn on day 4 were similar in the two groups (84% for rabeprazole; 95% confidence interval, 76-90%; 83% for omeprazole; 95% confidence interval, 75-89%). There were no significant differences between the treatments in the relief from other gastro-oesophageal reflux disease symptoms or in healing rates. The number of reports of severe heartburn during the first 3 days was higher in the omeprazole group (daytime heartburn: 4.7% for rabeprazole vs. 10.3% for omeprazole, P=0.005; night-time heartburn: 4.7% for rabeprazole vs. 9.8% for omeprazole, P=0.01; statistical comparisons defined post hoc). CONCLUSIONS: Standard-dose rabeprazole was as effective as high-dose omeprazole in relieving symptoms by day 4 of treatment and in healing oesophageal lesions, but had a faster onset of action in patients with severe heartburn. This suggests that the improved pharmacological properties of rabeprazole translate into a clinically relevant advantage. PMID- 11876702 TI - Helicobacter pylori and dyspepsia: physicians' attitudes, clinical practice, and prescribing habits. AB - BACKGROUND: Consensus guidelines have been published on the management of Helicobacter pylori infection and it is assumed that these guidelines are adhered to in clinical practice. AIM: To assess the changing attitudes of medical practitioners to H. pylori, and the impact of H. pylori infection on everyday clinical practice and prescribing patterns. METHODS: Data for this review were gathered up to December 2000 from detailed review of medical journals, the biomedical database MEDLINE, and relevant abstracts. RESULTS: Physician surveys show widespread acceptance of H. pylori as a causal agent in peptic ulcer disease. Gastroenterologists adopted H. pylori therapy for peptic ulcer earlier and more comprehensively than primary care physicians. Despite a low level of belief in H. pylori as a causal agent in non-ulcer dyspepsia and gastro oesophageal reflux disease (GERD), H. pylori therapy is widely prescribed for these conditions. Proton pump inhibitor-based triple therapy is the eradication regimen of choice by all physician groups. In routine clinical practice, there appears to be significant under-treatment of peptic ulcer disease with H. pylori therapy, but extensive use for non-ulcer indications. Prescription of H. pylori treatment regimens of doubtful efficacy appears commonplace, and are more likely in primary care. Despite the advent of H. pylori therapy, the prescription of antisecretory therapy, particularly of proton pump inhibitors, continues to rise. CONCLUSIONS: Publication of consensus guidelines per se is not enough to ensure optimal management of H. pylori infection. Innovative and ongoing educational measures are needed to encourage best practice in relation to H. pylori infection. These measures might be best directed at primary care, where the majority of dyspepsia is managed. PMID- 11876703 TI - Interactions between Helicobacter pylori and other risk factors for peptic ulcer bleeding. AB - AIM: To investigate the role of Helicobacter pylori, expressing the virulence marker CAGA (cytotoxin associated gene product A) in ulcer complications and its interaction with nonsteroidal anti-inflammatory drugs (NSAIDs) and other risk factors. DESIGN: Case control study using conditional logistic regression analysis. SETTING: University and City Hospitals, Nottingham. SUBJECTS: 203 consecutive patients with ulcer bleeding and 203 age- and sex-matched controls. RESULTS: Ulcer bleeding was more likely with positive H. pylori serology (odds ratio = 3.3, 95% CI: 1.7--6.6 for CagA positive, but only OR = 1.6, 95% CI: 0.7 3.7 for CagA negative serology), current smoking (OR 2.2, 95% CI: 1.04-4.7), aspirin < or = 300 mg daily (OR 7.7, 95% CI: 2.8-20.6), all other nonsteroidal anti-inflammatory drugs (NSAIDs: OR 10.6, 95% CI: 3.1-35.7 for < or = 1 defined daily dose lower and OR 22.6, 95% CI: 6.2-82.0 for higher doses) and past ulcer history (OR 5.6, 95% CI: 2.3-14.1). Aspirin < or = 300 mg daily was used by 25.1% of patients vs. 7.4% of controls. Smoking only enhanced risk in the presence of H. pylori, with a synergistic interaction (interaction odds ratio = 4.9, 2.4-9.9, P=0.002). Conversely, risks with non-aspirin NSAIDs were reduced in the presence of H. pylori, particularly if CagA-positive (interaction odds ratio=0.21, 0.05 0.9, P=0.03). CONCLUSIONS: CagA positive H. pylori infection is associated with an increased risk of ulcer bleeding. The risk from non-aspirin NSAIDs is even higher, but is less in H. pylori infected people. Low-dose aspirin is now commonly associated with ulcer bleeding. PMID- 11876704 TI - Role of Helicobacter pylori serology in atrophic body gastritis after eradication treatment. AB - BACKGROUND: It has been reported that 50% of patients with atrophic body gastritis have positive Helicobacter pylori antibody titres only. In atrophic body gastritis, a decrease in H. pylori antibodies after eradication treatment has been reported, suggesting that serology may indicate an active H. pylori infection. AIM: To investigate the time course of H. pylori antibodies and gastric inflammation after eradication treatment in patients with atrophic body gastritis, and to determine whether serology alone can be considered as a valid tool to assess the efficacy of eradication treatment in patients with atrophic body gastritis. METHODS: Twenty-seven patients with atrophic body gastritis (12 serologically H. pylori-positive only, ABG-S+; 15 H. pylori-positive at histology and serology, ABG-H+) were included in the treatment group, and 17 patients (all ABG-S+) in the no treatment group. All patients had gastroscopy plus biopsies evaluated according to the updated Sydney system and H. pylori immunoglobulin G determination: in the treatment group, at baseline and 6 and 24 months after eradication (bismuth-based triple regimens); in the no treatment group, at baseline and after 3 years. RESULTS: In the treatment group, in ABG-S+ patients, H. pylori antibodies decreased significantly 6 months after treatment [37.5 U/mL (16-100 U/mL) vs. 15 U/mL (0--100 U/mL), P < 0.01], but 2 years after treatment no further decrease occurred. In addition, in ABG-H+ patients, a significant decrease in H. pylori antibodies occurred 6 months after treatment [45 U/mL (12.5 100 U/mL) vs. 31 U/mL (0-65 U/mL), P < 0.01], but a further decrease was also observed 2 years after treatment [20 U/mL (0-56 U/mL), P < 0.01]. In ABG-S+ patients, no correlation was observed between the H. pylori antibodies and gastric inflammation score, whereas, in the ABG-H+ group, this correlation was extremely significant (r=0.5991, P < 0.0001). In the no treatment group, at follow-up, a significant decrease in H. pylori antibodies was observed [26 U/mL (15-100 U/mL) vs. 22 U/mL (0-53 U/mL), P < 0.05], but the gastric body inflammation remained unchanged. CONCLUSIONS: This study shows that, in ABG-S+ patients after eradication treatment, serology does not keep in step with gastric inflammation. This suggests that, in patients with atrophic body gastritis, serology alone may not be valid for the assessment of the efficacy of eradication treatment. PMID- 11876705 TI - The effect of Helicobacter pylori eradication on the natural course of atrophic gastritis with dysplasia. AB - BACKGROUND: There are few data on the natural course of Helicobacter pylori related atrophic gastritis. AIM: To investigate the effect of H. pylori eradication on advanced atrophic gastritis in the corpus. METHODS: Twenty-two elderly men with H. pylori infection and moderate or severe atrophic corpus gastritis formed the study population. These men were under endoscopic surveillance because of the presence of indefinite or definite dysplastic gastric lesions in addition to atrophic corpus gastritis. The men were gastroscopically and bioptically examined four times before they received H. pylori eradication therapy (mean follow-up time, 7.5 years), and once again 2.5 years after eradication therapy. Serum levels of pepsinogen I and H. pylori antibodies were analysed at baseline, immediately before and 2.5 years after eradication therapy. RESULTS: During the 7.5-year period prior to eradication therapy, no significant changes were observed in the mean atrophy and intestinal metaplasia scores or in the mean serum level of pepsinogen I. However, a significant improvement occurred in the mean histological scores of inflammation (from 2.2 to 0.5), atrophy (from 2.2 to 1.2) and intestinal metaplasia (from 1.6 to 1.1) in the corpus mucosa after H. pylori eradication. In addition, the mean serum level of pepsinogen I increased from 16.3 to 25.7 microg/L (P=0.0071, Wilcoxon signed rank test) after eradication therapy. CONCLUSIONS: The results suggest that advanced atrophic corpus gastritis (and intestinal metaplasia) improves and may even heal after the eradication of H. pylori. PMID- 11876706 TI - Optimal serological tests for the detection of Helicobacter pylori infection in the Chinese population. AB - BACKGROUND: Numerous serological tests for the detection of Helicobacter pylori infection have been developed. However, many perform poorly when evaluated in the Chinese population. AIM: To search for optimal serological tests for the detection of H. pylori infection in Chinese patients. METHODS: Consecutive dyspeptic patients referred for upper endoscopy were recruited. During endoscopy, gastric biopsies were taken for the CLOtest and histological examination. Patients were then given a 13C-urea breath test. Sera were used to test for H. pylori infection, employing three commercial enzyme-linked immunosorbent assay kits (pylori DTect, HP IgG and GAP IgG). Results were compared with the gold standard defined by the CLOtest, histology and 13C-urea breath test. RESULTS: Among the 142 patients (47 male, 95 female; mean age, 49 years) recruited, 81 (57%) were H. pylori-positive, 57 (40%) were H. pylori-negative and four (3%) were defined to be indeterminate. Using a self-defined cut-off value after calculation, the best accuracies for the pylori DTect, HP IgG and GAP IgG tests were 97%, 91% and 80%, respectively. CONCLUSIONS: The pylori DTect test is an optimal serological test for the detection of H. pylori infection in Hong Kong Chinese patients. The HP IgG test may be used as an alternative. PMID- 11876707 TI - Mono, dual and triple moxifloxacin-based therapies for Helicobacter pylori eradication. AB - BACKGROUND: Moxifloxacin is a broad spectrum fluoroquinolone with single daily administration, currently used, above all, for respiratory tract infections. AIM: To compare the efficacy of different 1-week moxifloxacin-based Helicobacter pylori eradication regimens. METHODS: One hundred and twenty H. pylori-positive subjects were randomized to receive moxifloxacin (400 mg/day), moxifloxacin (400 mg/day) and lansoprazole (30 mg/day) or moxifloxacin (400 mg/day), lansoprazole (30 mg/day) and clarithromycin (500 mg b.d.). H. pylori status was reassessed 6 weeks after the end of therapy, and both intention-to-treat and per protocol analyses were performed. RESULTS: One hundred and nineteen of the 120 patients completed the study. H. pylori eradication was achieved in 22.5% of patients treated with moxifloxacin, in 33.3% of subjects treated with moxifloxacin and lansoprazole and in 90% of patients treated with moxifloxacin, clarithromycin and lansoprazole. CONCLUSIONS: Mono and dual moxifloxacin-based therapies are not acceptable for H. pylori eradication; conversely, moxifloxacin-based triple therapy may be considered as a new, effective, first-line therapy option. PMID- 11876709 TI - Effect of Helicobacter pylori eradication on oesophageal acid exposure in patients with reflux oesophagitis. AB - BACKGROUND: The effect of Helicobacter pylori eradication on reflux oesophagitis is unclear. AIM: To study the effect of H. pylori eradication on oesophageal acid exposure and disease severity in patients with reflux oesophagitis. METHODS: Patients with reflux oesophagitis and H. pylori infection were recruited for 24-h oesophageal pH-metry. They were then randomly assigned to receive either treatment for H. pylori eradication (1-week omeprazole-based triple therapy, followed by 7-week omeprazole) or omeprazole alone (8-week omeprazole). Uninfected patients were recruited as controls. Endoscopy, pH monitoring and symptom assessment were repeated at 26 weeks. RESULTS: Forty patients (25 H. pylori-positive and 15 uninfected) with erosive oesophagitis were studied. Fourteen were randomized to receive treatment for H. pylori eradication and 11 to receive omeprazole alone. There was no difference in the percentage of time the oesophageal pH < 4 before and 26 weeks after treatment among the three groups. However, the percentage of time the oesophageal pH < 2 (P=0.01) and pH < 3 (P=0.02) was significantly increased in patients receiving treatment for H. pylori eradication. Three (21%) patients in the group receiving treatment for H. pylori eradication had worsening of reflux oesophagitis. CONCLUSIONS: H. pylori eradication increases oesophageal acid exposure and may adversely affect the clinical course of reflux disease in a subset of patients. PMID- 11876708 TI - Specific identification of three low molecular weight membrane-associated antigens of Helicobacter pylori. AB - BACKGROUND: A large number of Helicobacter pylori proteins are antigenic, but antibodies to these proteins persist in spite of the eradication of the infection. METHODS AND RESULTS: The analysis of sera from H. pylori-infected and non-infected patients, before and 3 and 5 months after eradication, showed that the antibody response against unknown H. pylori antigens at 32, 30, 22 and 14 kDa in sodium dodecylsulphate polyacrylamide gel electrophoresis decreased by > or = 60% at 3 months and > or = 70% at 5 months after treatment. Two-dimensional gel electrophoresis and mass spectrometry allowed the identification of eight proteins at these positions: neuraminyl-lactose-binding haemagglutinin precursor, 3-oxoadipate CoA-transferase subunit A, elongation factor P, peptidoglycan associated lipoprotein precursor, hypothetical protein HP0596, adhesin-thiol peroxidase, 50S ribosomal protein L7/L12 and subunit b' of the F(0) ATP synthase. Three of these eight, expressed as recombinant proteins (32 kDa neuraminyl lactose-binding haemagglutinin precursor, 30 kDa peptidoglycan-associated lipoprotein precursor and 22 kDa hypothetical protein HP0596), reacted specifically with sera from infected patients, while the 14 kDa 50S ribosomal protein L7/L12 cross-reacted with one out of five sera from H. pylori-negative patients. The other recombinant proteins did not show significant immunoreactivity. CONCLUSIONS: Four low molecular weight antigens were identified by these methods, three of which were specific. Immunoreaction with these three proteins (neuraminyl-lactose-binding haemagglutinin precursor, peptidoglycan associated lipoprotein precursor and hypothetical protein HP0596) could provide a serological assessment not only of H. pylori infection, but also of eradication. PMID- 11876710 TI - Different effects of short-term omeprazole, lansoprazole or pantoprazole on the accuracy of the (13)C-urea breath test. AB - BACKGROUND: Proton pump inhibitors may interfere with the accuracy of the (13)C urea breath test, but little information is available on the effect of standard doses of various proton pump inhibitors on this test. AIM: To evaluate the effect of short-term standard doses of omeprazole, lansoprazole and pantoprazole on the accuracy of the standardized (13)C-urea breath test. METHODS: A total of 124 patients with Helicobacter pylori infection, diagnosed on the basis of gastric histology, rapid urease test and (13)C-urea breath test, were studied. These patients received omeprazole, 20 mg/day, lansoprazole, 30 mg/day, or pantoprazole, 40 mg/day, for 2 weeks according to a randomized protocol. (13)C Urea breath test was repeated on days 4, 7 and 14 while on therapy and 7 days after proton pump inhibitor withdrawal. RESULTS: Of the patients receiving omeprazole and lansoprazole, 30% (12/40) and 20% (8/41), respectively, became (13)C-urea breath test negative during therapy, compared with none of the 42 patients treated with pantoprazole (P < 0.05). All the false negative (13)C-urea breath test results returned to positive within 1 week of drug withdrawal, with a mean recovery to 84.7 +/- 15.6% of baseline delta(13)CO(2). CONCLUSIONS: Short term omeprazole and lansoprazole interfere with the 13C-urea breath test, although a return to positive test results invariably occurs within 1 week of proton pump inhibitor withdrawal. In contrast, the accuracy of the (13)C-urea breath test does not appear to be significantly impaired by short-term pantoprazole, which therefore may not necessarily be withdrawn before this test. PMID- 11876711 TI - The cost-effectiveness of population Helicobacter pylori screening and treatment: a Markov model using economic data from a randomized controlled trial. AB - BACKGROUND: Economic models have suggested that population Helicobacter pylori screening and treatment may be a cost-effective method of reducing mortality from gastric cancer. These models are conservative as they do not consider that the programme may reduce health service peptic ulcer and other dyspepsia costs. We have evaluated the economic impact of population H. pylori screening and treatment over 2 years in a randomized controlled trial and have incorporated the results into an economic model exploring the impact of H. pylori eradication on peptic ulcer disease and gastric cancer. METHODS: Subjects between the ages of 40 and 49 years were randomly invited to attend their local primary care centre. H. pylori status was evaluated by (13)C-urea breath test and infected individuals were randomized to receive omeprazole, 20 mg b.d., clarithromycin, 250 mg b.d., and tinidazole, 500 mg b.d., for 7 days or identical placebos. Economic data on health service costs for dyspepsia were obtained from a primary care note review for the 2 years following randomization. These data were incorporated into a Markov model comparing population H. pylori screening and treatment with no intervention. RESULTS: A total of 2329 of 8407 subjects were H. pylori positive: 1161 were randomized to receive eradication therapy and 1163 to receive placebo. The cost difference favoured the intervention group 2 years after randomization, but this did not reach statistical significance (11.42 ponds sterling per subject cost saving; 95% confidence interval, 30.04 ponds sterling to -7.19 pounds sterling; P=0.23). Analysis by gender suggested a statistically significant dyspepsia cost saving in men (27.17 ponds sterling per subject; 95% confidence interval, 50.01 pounds sterling to 4.32 pounds sterling; P=0.02), with no benefit in women (-4.46 per subject; 95% confidence interval, -33.85 pounds sterling to 24.93 pounds sterling). Modelling of these data suggested that population H. pylori screening and treatment for 1,000,000 45-year-olds would save over 6,000,000 pounds sterling and 1300 years of life. The programme would cost 14, 200 pounds sterling per life year saved if the health service dyspepsia cost savings were the lower limit of the 95% confidence intervals and H. pylori eradication had only a 10% efficacy in reducing mortality from distal gastric cancer and peptic ulcer disease. CONCLUSIONS: Modelling suggests that population H. pylori screening and treatment are likely to be cost-effective and could be the first cost-neutral screening programme. This provides a further mandate for clinical trials to evaluate the efficacy of population H. pylori screening and treatment in preventing mortality from gastric cancer and peptic ulcer disease. PMID- 11876712 TI - Effectiveness of two quadruple, tetracycline- or clarithromycin-containing, second-line, Helicobacter pylori eradication therapies. AB - BACKGROUND: There are no guidelines on second-line therapies for Helicobacter pylori eradication failures of omeprazole-clarithromycin-amoxicillin triple therapy. AIM: To compare the efficacy of two second-line therapies for persistent H. pylori infection. METHODS: Over a 6-year period, patients with persistent H. pylori infection following omeprazole-clarithromycin-amoxicillin eradication therapy were randomized to receive omeprazole, 20 mg twice daily, bismuth, 120 mg four times daily, metronidazole, 500 mg twice daily, and either tetracycline, 500 mg four times daily, or clarithromycin, 500 mg twice daily, given for 7 days. Before therapy, patients underwent endoscopy with biopsies for histology, culture and antibiotic susceptibility tests. H. pylori infection was confirmed by histology. RESULTS: Of the 95 randomized patients, 88 (93%) completed the study. Age, sex, smoking, ulcer/non-ulcer dyspepsia ratio and antibiotic resistance were not significantly different between the treatment groups. On intention-to-treat analysis, eradication was achieved in 41 of the 49 patients (84%; 95% confidence interval, 70.4-92.7%) and 27 of the 46 patients (59%; 95% confidence interval, 43.3-73.0%) of the tetracycline- and clarithromycin-containing groups, respectively (P=0.007). On multivariate regression analysis, the sensitivity of H. pylori to metronidazole had a likelihood ratio of 5.2 (P=0.022), followed by the type of quadruple therapy (likelihood ratio, 4.4; P=0.036). CONCLUSIONS: Tetracycline-containing quadruple rescue therapy is highly effective in treating H. pylori eradication failures of the omeprazole-amoxicillin-clarithromycin regimen. PMID- 11876713 TI - Influence of Helicobacter pylori infection and eradication on blood lipids and fibrinogen. AB - BACKGROUND: An association between Helicobacter pylori infection and heart disease has been suggested. A potential mechanism may be inflammation-induced atherogenic changes of lipoproteins, but epidemiological studies have provided conflicting results. METHODS: In a prospective multicentre study, 830 patients submitted for endoscopy and H. pylori testing were evaluated. Of the 686 H. pylori-positive patients, 487 received and 199 did not receive eradication treatment. Serum lipids and plasma fibrinogen were measured at baseline in all patients and 3 months later in those initially positive for H. pylori. RESULTS: H. pylori had no influence on baseline lipid or fibrinogen levels. Increases in high-density lipoprotein cholesterol were observed in 368 patients who received eradication treatment and in 193 untreated patients: 0.06 mmol/L (P=0.000) and 0.07 mmol/L (P=0.009), respectively. Similar minor increases in total cholesterol and triglycerides occurred in both groups. Lipid changes were related to symptom relief and a reduction in smoking. Eradication therapy was associated with a minor decrease in plasma fibrinogen irrespective of the resolution of infection. CONCLUSIONS: H. pylori has no influence on blood lipids or fibrinogen. Both the eradication of infection and symptomatic treatment without eradication are associated with minor lipid changes related to symptom relief and lifestyle modifications. Thus, the inflammatory changes associated with H. pylori are unlikely to affect lipoprotein or fibrinogen metabolism. PMID- 11876714 TI - Bifidobacterium animalis strain DN-173 010 shortens the colonic transit time in healthy women: a double-blind, randomized, controlled study. AB - BACKGROUND: A previous study has suggested that Bifidobacterium animalis DN-173 010 shortens the colonic transit time in women. AIM: To confirm this effect and to determine whether modifications of the faecal bacterial mass and/or faecal secondary bile salts may be the explanation. METHODS: A double-blind, cross-over study was performed. Thirty-six healthy women were studied in four consecutive 10 day periods. During periods 2 and 4, they ingested three 125 g cups per day of a fermented milk which was either a product containing B. animalis DN-173 010 or a control without bifidobacteria. Periods 1 and 3 were run-in and washout periods, respectively. The total and segmental colonic transit times were assessed using a pellet method. In 12 subjects, all stools were collected and analysed for pH, faecal weight, bacterial mass and bile acids. RESULTS: The total and sigmoid transit times were significantly shorter during dosing with B. animalis compared to the control period. The other transit times, faecal weight, pH, bacterial mass and bile acids were not significantly affected. CONCLUSIONS: B. animalis DN-173 010 shortens the colonic transit time in healthy women. This effect is not explained by modifications of the faecal bacterial mass or secondary bile acids. PMID- 11876715 TI - Albendazole chemotherapy for AIDS-related diarrhoea in Zambia--clinical, parasitological and mucosal responses. AB - BACKGROUND: Albendazole reduces diarrhoea in African AIDS patients, but it is unclear if the clinical response to treatment reflects pathogen eradication and/or mucosal recovery. METHODS: Adults with HIV-related persistent diarrhoea were treated with albendazole 800 mg twice daily for 14 days. Clearance of parasites was evaluated at 3 and 6 weeks by stool microscopy. At baseline and at 6 weeks duodenal biopsies were taken for electron microscopy (EM) and morphometry. RESULTS: Ten (7%) of 153 patients had cryptosporidiosis, 54 (37%) had isosporiasis and 23 (16%) had microsporidiosis. By 3 weeks, these protozoa were cleared in 27 (46%) of 59 patients initially positive. By 6 weeks, 34 (39%) of 87 patients experienced complete clinical response, 18 (21%) partial response and 35 (40%) no response. Crypt depth increased by 15% over 6 weeks (P < 0.001), but villous height increased only in patients with complete response (median + 50 microm, interquartile range (IQR) 2-90, compared to patients with partial (+ 4 microm, IQR -15,41) or no response (-13 microm, IQR -2,12; P=0.008)). Fifteen patients died: body mass index < 17.5 kg/m(2) and crypt depth < 180 microm independently predicted death. CONCLUSIONS: Albendazole therapy reduced the burden of protozoal infection and promoted mucosal recovery in patients with a complete clinical response. PMID- 11876716 TI - The effects of the specific 5HT(4) receptor agonist, prucalopride, on colonic motility in healthy volunteers. AB - BACKGROUND: Prucalopride is a selective and specific 5-hydroxytryptamine(4) receptor agonist that is known to increase stool frequency and to accelerate colonic transit. AIM: To investigate the effect of prucalopride on high-amplitude propagated contractions and segmental pressure waves in healthy volunteers. METHODS: After 1 week of dosing (prucalopride or placebo in a double-blind, randomized, crossover fashion), colonic pressures were recorded in 10 healthy subjects using a solid-state pressure catheter with six sensors spaced 10 cm apart. Subjects kept diary records of their bowel habits (frequency, consistency and straining). High-amplitude propagated contractions were analysed visually, comparing their total numbers and using 10-min time windows. Segmental pressure waves were analysed using computer algorithms, quantifying the incidence, amplitude, duration and area under the curve of all detected peaks. RESULTS: When taking prucalopride, stool frequency increased, consistency decreased and subjects strained less. Prucalopride just failed to increase the total number of high-amplitude propagated contractions (P=0.055). The number of 10-min time windows containing high-amplitude propagated contractions was increased by prucalopride (P=0.019). Prucalopride increased the area under the curve per 24 h (P=0.026). CONCLUSIONS: The 5-hydroxytryptamine(4) receptor agonist prucalopride stimulates high-amplitude propagated contractions and increases segmental contractions, which is likely to be the underlying mechanism of its effect on bowel habits in healthy volunteers. PMID- 11876717 TI - Effect of acute and chronic levosulpiride administration on gastric tone and perception in functional dyspepsia. AB - BACKGROUND: Altered visceral perception is common in functional dyspepsia (FD). Dopaminergic pathways control gastrointestinal motility, but whether they modulate visceral sensitivity is unknown. AIM: To investigate whether levosulpiride, a D2 antagonist, modulates gastric sensitivity and compliance in dyspeptic patients. METHODS: Eight healthy subjects and 16 dyspeptic patients underwent graded gastric distensions using a tensostat. In dyspeptic patients the same isotonic distensions were repeated during either levosulpiride or saline administration. Eight FD patients were evaluated after 4-week treatment with oral levosulpiride. Gastrointestinal symptoms were evaluated using a 100 mm visual analogue score. Perception was scored on a scale of 0 to 6. RESULTS: Although healthy subjects and FD patients had similar gastric compliance, FD patients tolerated lower tension levels. At the same distending tension levels, levosulpiride decreased gastric compliance and perception score (14 +/- 6% and 38 +/- 10% change, respectively; P < 0.05 vs. saline) only in FD patients. Isotonic distensions exhibited very reproducible perception. Chronic levosulpiride administration significantly reduced dyspeptic symptoms and increased discomfort threshold. CONCLUSIONS: Compared with healthy subjects, FD patients show marked gastric hypersensitivity. In FD patients levosulpiride decreased the perception of gastric distension with an action unrelated to change of gastric tone. Chronic levosulpiride administration significantly ameliorates gastrointestinal symptoms and increases the discomfort threshold. PMID- 11876718 TI - Effect of granisetron on radiation-induced alterations of colonic motility and fluid absorption in rats. AB - BACKGROUND: Radiation-induced diarrhoea is attributed to both mucosal injury and alterations of intestinal motility. Previous reports have indicated that 5 hydroxytryptamine is released following irradiation, which may contribute to these changes. AIMS: To investigate the effects of granisetron (5 hydroxytryptamine type-3 receptor antagonist) on colonic motility, fluid absorption and 5-hydroxytryptamine colonic content following abdominal irradiation (10 Gy) in rats. METHODS: In vivo measurements of motility and fluid absorption in the proximal and distal colon, respectively, diarrhoea score and 5 hydroxytryptamine tissue levels were performed 3 and 7 days after abdominal irradiation. The effects of post-irradiation granisetron (0.3 mg/kg subcutaneously) were also evaluated. RESULTS: Colonic motility and fluid/Na(+) absorption were reduced 3 and 7 days after irradiation. All rats developed diarrhoea (3 days) concomitant with increased colonic mucosal 5-hydroxytryptamine levels. Granisetron prevented diarrhoea, attenuated decreased colonic motility and reduced 5-hydroxytryptamine levels on day 3, although fluid absorption was only slightly improved. On day 7, colonic motility and fluid/Na(+) absorption were restored in granisetron-dosed animals. CONCLUSIONS: These results indicate that radiation-induced diarrhoea involves alterations of both colonic motility and fluid/Na(+) absorption. 5-Hydroxytryptamine could be one of the mediators implicated in these alterations, as post-irradiation dosage with a 5 hydroxytryptamine type-3 receptor antagonist improved colonic motility and prevented diarrhoea on day 3. PMID- 11876719 TI - Lectins can reverse the distal intestinal atrophy associated with elemental diets in mice. AB - BACKGROUND: Elemental diets cause intestinal atrophy and reduced intestinal integrity, which can lead to significant increases in intestinal permeability and bacterial translocation. Recently, several lectins have been shown to have trophic effects on the intestine. AIMS: We examined the effects of concanavalin-A and phytohaemagglutinin on cell proliferation and crypt fission throughout the intestine of mice fed on elemental diets. METHODS: Mice were randomized to chow fed, elemental diet, elemental diet plus concanavalin-A and elemental diet plus phytohaemagglutinin groups. Cell proliferation and crypt fission were estimated in microdissected crypts. Plasma gastrin and enteroglucagon levels were measured by radioimmunoassay. RESULTS: Elemental diet feeding significantly decreased cell proliferation and crypt fission of the middle and distal small intestine and throughout the colon. Phytohaemagglutinin significantly increased the weight of the intestine, but concanavalin-A had little effect. Cell proliferation in the small intestine was significantly increased by both lectins. However, in the stomach and colon, only phytohaemagglutinin increased proliferation. Crypt fission in the colon was dramatically increased by phytohaemagglutinin. Phytohaemagglutinin increased the plasma gastrin level, but not the enteroglucagon level. CONCLUSIONS: Lectins have significant trophic effects on the small intestine and colon of mice fed elemental diets, and these actions vary between different sites in the gastrointestinal tract. PMID- 11876720 TI - The cathartic colon? PMID- 11876721 TI - H. pylori antimicrobial resistance. PMID- 11876723 TI - Drugs for bleeding peptic ulcer. PMID- 11876724 TI - UROLINK: a model for working together in a changing world. PMID- 11876725 TI - UROLINK in sub-Saharan Africa. PMID- 11876726 TI - UROLINK--benefits for trainees from both sides. PMID- 11876727 TI - Twinning: the future for sustainable collaboration. PMID- 11876728 TI - Urology in Eastern Europe in the 1990s. PMID- 11876729 TI - Bilharziasis of the genitourinary tract. PMID- 11876730 TI - Reconstructive urology in the tropical and developing world: a personal perspective. PMID- 11876732 TI - Clinical nephrological problems important to the urologist. PMID- 11876731 TI - The role of the urologist in the treatment and elimination of lymphatic filariasis worldwide. PMID- 11876733 TI - Providing an obstetric fistula service. PMID- 11876734 TI - Free bladder mucosal autograft in the treatment of complicated vesicovaginal fistula. AB - OBJECTIVE: To describe an effective and simple method for repairing complicated vesicovaginal fistula using a free bladder mucosal autograft. PATIENTS AND METHODS: The study included 14 patients with an iatrogenic vesicovaginal fistula after hysterectomy or prolonged obstructed labour. An initial fistula repair had been attempted in 12 patients. All patients were then repaired using a free bladder mucosal graft technique 3-24 months (mean 8) after the fistula was diagnosed. RESULTS: Of the 14 patients 12 had an immediately successful result, with no evidence of leakage, but two required prolonged catheter drainage. Thirteen patients remained dry at the follow-up 3-15 months later. Two patients had stress urinary incontinence and three patients had urinary tract infection. CONCLUSIONS: The free bladder mucosal autograft technique is a simple, effective and fast method for vesicovaginal fistula repair. The technique produces an excellent repair of complicated vesicovaginal fistula, and appears to be more physiological and easier to perform than other transabdominal techniques. PMID- 11876735 TI - Retrospective review and long-term follow-up of radical cystectomy in a developing country. AB - OBJECTIVE: To retrospectively review the clinical data from patients undergoing radical cystectomy for bladder cancer, and to analyse the complications and survival rates associated with this operation in a developing country. PATIENTS AND METHODS: The study comprised 105 patients who underwent radical cystectomy from 1986 to 1993. Data were collected from retrospective reviews of hospital and physician's office records, and by contact with the patients. Metastatic status was evaluated before surgery and tumours staged using the Tumour-Nodes-Metastasis classification. The indication for surgery was histologically confirmed muscle invasion after transurethral resection biopsy, or endoscopically uncontrollable superficial disease. The data were analysed to assess the perioperative complications and long-term survival, with 5-year survival rates determined using Kaplan-Meier survival curves. RESULTS: The complication rate was 27.6%; most of the complications were managed conservatively with good results and re-operation was required in only two patients. There were two deaths (1.9%) at 15-45 days after surgery, but none during surgery. Patients were divided into node-negative and node-positive groups for analysis and 5-year survival rates determined; for node-negative organ-confined disease (< or =pT3A) the survival was 68% and for nonorgan-confined disease (> or =pT3B) 25%. The 5-year survival rate in the presence of nodal metastases was 13% for N1 and none for N2 disease. Six patients developed urethral recurrence, detected on follow-up urethral-wash cytology. Five of these patients underwent urethrectomy, and four of the six survived for 5 years. Pelvic recurrence occurred in five patients (4.7%), none of whom survived for 5 years. CONCLUSION: Radical cystectomy and pelvic lymph node dissection remains the mainstay of treatment in muscle-invasive bladder cancer. This is a relatively safe procedure with minimal morbidity and mortality; 68% of the present patients with organ-confined disease survived 5 years and 12 patients were alive at 10 years, indicating the effectiveness of this operation in selected cases. However, < 29% of patients with nonorgan-confined and nodal metastatic disease survived 5 years, thereby implying the need for more effective adjuvant therapy in these patients. Radical cystectomy is a viable option in developing countries, with 5-year survival rates comparable with most large published series. PMID- 11876736 TI - The management of stone disease. PMID- 11876738 TI - Helping to understand studies examining genetic susceptibility to sepsis. PMID- 11876740 TI - Immune response modulation by recombinant peptides expressed in virus-like particles. AB - Aspergillus fumigatus, a ubiquitous fungus, is implicated in the pathogenesis of a number of clinically different allergic diseases in man, including allergic bronchopulmonary aspergillosis. Peptide-based immunotherapy may offer an alternative treatment strategy for the management of allergic disease. The objective of this study was to alter the allergen-specific immune response using dominant T cell epitopes of a major A. fumigatus allergen, Asp f2, expressed in yeast as virus-like particles (VLP). The T cell epitopes of Asp f2, recognized in mice with an H-2d background, were determined by producing T-cell hybridomas. Two dominant T cell epitopes, aa60--71 and aa235--249, were identified and expressed in a yeast VLP system. To induce tolerance VLP-peptides were injected subcutaneously into mice previously immunized with recombinant Asp f2. The T cell immune response was abrogated totally in 3 weeks following a single injection of VLP but was restored 2 months later following intranasal antigen exposure. T-cell depletion resulted in the reduction of 20-30% of all antigen-specific immunoglobulin classes. Thus, recombinant peptides expressed in the VLP system can be used successfully in the modulation of Asp f2-induced immune response in mice, although a single administration is not sufficient to maintain a state of tolerance for a long period of time. PMID- 11876739 TI - Unlocking the immunological mechanisms of orbital inflammation in thyroid eye disease. PMID- 11876741 TI - Improved outcome of chronic Pseudomonas aeruginosa lung infection is associated with induction of a Th1-dominated cytokine response. AB - Repeated challenge with antigen is involved in the pathogenesis of a variety of pulmonary diseases. Patients with cystic fibrosis (CF) experience recurrent pulmonary colonization with Pseudomonas aeruginosa before establishment of chronic lung infection. To mimic recurrent lung infections in CF patients, the lungs of susceptible BALB/c mice were re-infected with P. aeruginosa 14 days after the initial infection. Singly-infected BALB/c mice, as well as non-infected mice, were used as controls. Decreased mortality and milder lung inflammation in re-infected BALB/c mice, as well as a tendency for improved clearance of bacteria, was observed when compared with singly-infected mice. The improved outcome in re-infected mice correlated with changes in CD4 cell numbers. Surface expression of LFA-1 on pulmonary CD4 cells was increased in re-infected compared with singly-infected mice. Moreover, resistance to re-infection was paralleled by a shift towards a Th1-dominated response and increased IL-12 production. No significant increase in serum IgG was observed in the re-infected mice. In conclusion, these results indicate a protective role for a Th1-dominated response, independent of antibody production, in chronic P. aeruginosa lung infection in CF. PMID- 11876742 TI - Dendritic cells exposed in vitro to TGF-beta1 ameliorate experimental autoimmune myasthenia gravis. AB - Experimental autoimmune myasthenia gravis (EAMG) is an animal model for human myasthenia gravis (MG), characterized by an autoaggressive T-cell-dependent antibody-mediated immune response directed against the acetylcholine receptor (AChR) of the neuromuscular junction. Dendritic cells (DC) are unique antigen presenting cells which control T- and B-cell functions and induce immunity or tolerance. Here, we demonstrate that DC exposed to TGF-beta1 in vitro mediate protection against EAMG. Freshly prepared DC from spleen of healthy rats were exposed to TGF-beta1 in vitro for 48 h, and administered subcutaneously to Lewis rats (2 x 10(6)DC/rat) on day 5 post immunization with AChR in Freund's complete adjuvant. Control EAMG rats were injected in parallel with untreated DC (naive DC) or PBS. Lewis rats receiving TGF-beta1-exposed DC developed very mild symptoms of EAMG without loss of body weight compared with control EAMG rats receiving naive DC or PBS. This effect of TGF-beta1-exposed DC was associated with augmented spontaneous and AChR-induced proliferation, IFN-gamma and NO production, and decreased levels of anti-AChR antibody-secreting cells. Autologous DC exposed in vitro to TGF-beta1 could represent a new opportunity for DC-based immunotherapy of antibody-mediated autoimmune diseases. PMID- 11876743 TI - Antigen presentation by macrophages is enhanced by the uptake of necrotic, but not apoptotic, cells. AB - The aim of this study was to determine whether phagocytosis of necrotic or apoptotic cells affects antigen presentation by murine bone marrow-derived macrophages. After uptake of necrotic neutrophils, macrophages were able to stimulate significantly higher T cell proliferation in vitro against both the recall antigen albumin and the mitogen concanavalin A. No such effect was seen following phagocytosis of apoptotic neutrophils. Flow cytometry revealed that, within 4h of ingestion, macrophages that had taken up the necrotic cells expressed higher levels of CD40 than those that had phagocytosed apoptotic cells. Macrophage cultures pulsed with apoptotic, but not necrotic, neutrophils contained higher levels of transforming growth factor beta1, but lower concentrations of tumour necrosis factor alpha, compared to untreated controls. Our interpretation of these results is that macrophages that have taken up necrotic neutrophils co-stimulate T cells with greater efficiency due to rapid CD40 up-regulation, whereas those that have ingested apoptotic cells are not only ineffective in co-stimulation, but also secrete inhibitory cytokine. PMID- 11876744 TI - Lidocaine inhibits secretion of IL-8 and IL-1beta and stimulates secretion of IL 1 receptor antagonist by epithelial cells. AB - Lidocaine and related local anaesthetics have been shown to be effective in the treatment of ulcerative colitis (UC). However, the mechanisms underlying their therapeutic effect are poorly defined. Intestinal epithelial cells play an important role in the mucosal inflammatory response that leads to tissue damage in UC via the secretion of pro-inflammatory cytokines and chemokines. The aim of this study was to evaluate the direct immunoregulatory effect of lidocaine on pro inflammatory cytokine and chemokine secretion from intestinal epithelial cells. HT-29 and Caco-2 cell lines were used as a model system and treated with lidocaine and related drugs. The expression of IL-8, IL-1beta and the IL-1 receptor antagonist (RA) were assessed by ELISA and quantification of mRNA. In further experiments, the effect of lidocaine on the secretion of IL-8 from freshly isolated epithelial cells stimulated with TNFalpha was tested. Lidocaine, in therapeutic concentrations, inhibited the spontaneous and TNFalpha-stimulated secretion of IL-8 and IL-1beta from HT-29 and Caco-2 cell lines in a dose dependent manner. Similarly, suppression of IL-8 secretion was noted in the freshly isolated epithelial cells. Other local anaesthetics, bupivacaine and amethocaine, had comparable effects. Lidocaine stimulated the secretion of the anti-inflammatory molecule IL-1 RA. Both the inhibitory and the stimulatory effects of lidocaine involved regulation of transcription. The results imply that the therapeutic effect of lidocaine may be mediated, at least in part, by its direct effects on epithelial cells to inhibit the secretion of proinflammatory molecules on one hand while triggering the secretion of anti-inflammatory mediators on the other. PMID- 11876745 TI - Transfection of single-stranded hepatitis A virus RNA activates MHC class I pathway. AB - Although infection of single-stranded RNA viruses can enhance expression of major histocompatibility complex (MHC) class I genes, the mechanism underlying this process remains unclear. Recent studies have indicated that exposure of non immune cells to double-stranded deoxyribonucleic acid (DNA) or ribonucleic acid (RNA) of viral origin can directly increase the expression of MHC class I and related molecules without immune cell interaction. In this report, we show that transfection of single-stranded hepatitis A virus RNA into cultured hepatocytes results in the induction of genes for MHC class I, LMP2 and transporter for antigen processing (TAP1), in addition to the generation of viral proteins. We suggest that this stimulatory effect is due to the double-stranded RNA formed during replication of single-stranded viral RNA, and involves both double stranded, RNA-dependent protein kinase PKR and the secretion of IFNbeta. PMID- 11876746 TI - CD14-dependent and -independent cytokine and chemokine production by human THP-1 monocytes stimulated by Streptococcus suis capsular type 2. AB - Streptococcus suis capsular type 2 is an important aetiologic agent of swine meningitis, and it has been highlighted as a cause of occupational disease leading to meningitis and fulminant sepsis in humans. The objective of the present work was to study the ability of S. suis type 2 to induce the release of tumour necrosis factor alpha (TNF-alpha), interleukin-1 (IL-1), IL-6, IL-8 and monocyte chemotactic protein one (MCP-1) by human monocytic THP-1 cells. The induction of these five cytokines was dose- and incubation time-dependent, and it was significantly enhanced by pre-treatment of cells with interferon gamma. IL-8 levels were markedly higher compared with those obtained with the other cytokines. However, elevated levels of MCP-1 and IL-6 were also observed. Levels of cytokine induced by heat-killed or live bacteria were similar. Pre-treatment of cells with anti-CD14 monoclonal antibodies suggested that this important host receptor is partially implicated in TNF, IL-1, IL-6 and MCP-1 production, while CD14-independent pathways seem to be responsible for IL-8 production after S. suis stimulation. In addition, blocking studies with anti-TNF and anti-IL-1 antibodies revealed that these cytokines are involved in amplification of the S. suis-induced cytokine cascade. When several different S. suis strains of human or porcine origin were compared, a very heterogeneous pattern of cytokine production was observed. Human strains did not exhibit a clear tendency to induce higher cytokine release by human THP-1 monocytes. The synergistic effect of the up regulation of cytokines during S. suis meningitis may mediate many of the inflammatory reactions, including the sequestration of leucocytes at the site of infection. PMID- 11876747 TI - Myelin protein P0-specific IgM producing monoclonal B cell lines were established from polyneuropathy patients with monoclonal gammopathy of undetermined significance (MGUS). AB - Monoclonal expansion of B cells and plasma cells, producing antibodies against 'self' molecules, can be found not only in different autoimmune diseases, such as peripheral neuropathy (PN), but also in malignancies, such as Waldenstrom's macroglobulinaemia and B-type of chronic lymphocytic leukaemia (B-CLL), as well as in precancerous conditions including monoclonal gammopathy of undetermined significance (MGUS). About 50% of patients with PN-MGUS have serum antibodies against peripheral nerve myelin, but the specific role of these antibodies remains uncertain. The aims of the study were to establish, and characterize, myelin-specific B cell clones from peripheral blood of patients with PN-MGUS, by selection of cells bearing specific membrane Ig-receptors for myelin protein P0, using beads coated with P0. P0-coated magnetic beads were used for selection of cells, which subsequently were transformed by Epstein--Barr virus. The specificity of secreted antibodies was tested by ELISA. Two of the clones producing anti-P0 antibodies were selected and expanded. The magnetic selection procedure was repeated and new clones established. The cells were CD5+ positive, although the expression declined in vitro over time. The anti-P0 antibodies were of IgM-lambda type. The antibodies belonged to the VH3 gene family with presence of somatic mutations. The IgM reacted with P0 and myelin-associated glycoprotein (MAG), and showed no evidence for polyreactivity, in contrast to other IgM CD5+ clones included in the study as controls. The expanded clones expressed CD80 and HLA-DR, which is compatible with properties of antigen-presenting cells. The immunomagnetic selection technique was successfully used for isolation of antimyelin protein P0-specific clones. The cell lines may provide useful tools in studies of monoclonal gammopathies, leukaemia, and autoimmune diseases, including aspects of antigen-presentation by these cells followed by T cell activation. PMID- 11876748 TI - Placentally derived prostaglandin E2 acts via the EP4 receptor to inhibit IL-2 dependent proliferation of CTLL-2 T cells. AB - A number of immunomodulatory molecules are present in the placenta, including cytokines, prostaglandins, progesterone and indoleamine 2,3-dioxygenase. An undefined factor capable of down-regulating T-cell activity has recently been reported [1] as being produced by short-term cultures of placental fragments. By careful repetition of these studies we have confirmed that chorionic villi isolated from term placenta produce a low molecular weight, heat stable factor capable of inhibiting the IL-2-dependent proliferation of mouse CTLL-2 cells. This activity was not due, however, to a previously unknown immunosuppressive molecule, but rather to prostaglandin E2 (PGE2). Expression of cyclooxygenase (COX)-2 was detected in the syncytiotrophoblast of chorionic villi explants using immunohistochemistry. Culture of the explants in the presence of the COX-1/COX--2 inhibitors indomethacin and diclofenac, or with the COX-2-selective inhibitor DFP, blocked the production of the immunosuppressive factor. The immunosuppressive activity was restored by adding PGE2 to the supernatants obtained from diclofenac-inhibited explants. A number of different receptors are involved in mediating the biological effects of prostaglandins. By utilizing selective antagonists of individual receptors, we have established that the immunosuppressive effect of PGE2 on CTLL-2 cells is exerted via the EP4 receptor. Thus, addition of an EP4-selective antagonist, but not of EP1 or EP3 antagonists, abolished the immunosuppressive effect of PGE2 on CTLL-2 cells. This may have implications for attempts to selectively manipulate T-cell responses. PMID- 11876750 TI - IgE is expressed on, but not produced by, fetal cells in the human placenta irrespective of maternal atopy. AB - The prevalence of atopic diseases in children has increased during the last decades. Atopic symptoms usually appear early in life. This implies an early priming for atopic disease, possibly even at the fetal level. We therefore compared the presence and production of IgE in the local in utero environment during pregnancy in atopic and non-atopic women. Eighty-six women were included in the study. Fifty women were demonstrated to be atopics, based on clinical symptoms of atopic disease together with a positive Phadiatop and/or skin prick test. Placentas from these term pregnancies were obtained. Slices covering the full thickness of the placenta were cut clockwise around the umbilical cord and were analysed with immunohistochemistry. Surprisingly, numerous IgE+ cells, located primarily in the fetal villous stroma, were detected in a majority of the investigated placentas irrespective of the atopy of the mother or maternal or fetal total serum IgE levels. The placental IgE could not be demonstrated to be bound to IgE receptors, but was shown to be bound to fetal macrophages, possibly via FcgammaRI. No evidence was found for local fetal IgE production, although cells producing epsilon transcripts were occasionally detected in the decidua. We describe here the novel finding of numerous IgE+ cells in the human placenta, suggesting an hitherto unknown role for IgE in a successful pregnancy outcome, irrespective of whether or not the mother is atopic. PMID- 11876749 TI - Serum macrophage-derived chemokine (MDC) levels are closely related with the disease activity of atopic dermatitis. AB - Atopic dermatitis (AD) is a chronic and relapsing inflammatory skin disease characterized by the predominant infiltration of T cells, eosinophils and macrophages in lesional skin. Recently, macrophage-derived chemokine (MDC)/CCL22, a CC chemokine, was identified as a selective chemoattractant for CC chemokine receptor 4 (CCR4)-expressing cells, in addition to thymus and activation regulated chemokine (TARC). We have previously reported that serum TARC levels correlate with the severity of AD. In this report, we investigated the participation of MDC in AD. First, we measured serum MDC levels in 45 patients with AD, 25 patients with psoriasis vulgaris and 25 healthy controls. Serum MDC levels in AD patients were significantly higher than those in healthy controls and psoriasis patients. Furthermore, the increases in serum MDC levels in AD patients were greater in the severely affected group than in the moderate or mild groups. We compared serum MDC levels in 11 AD patients, before and after treatment, and observed a significant decrease after treatment. Moreover, the serum MDC levels significantly correlated with the Scoring AD (SCORAD) index, serum soluble (s) E-selectin levels, serum soluble interleukin-2 receptor (sIL 2R) levels, serum TARC levels and eosinophil numbers in peripheral blood. Our study strongly suggests that serum MDC levels have a notable correlation with disease activity and that MDC, as well as the CC chemokine TARC, may be involved in the pathogenesis of AD. PMID- 11876751 TI - Increased levels of serum tissue inhibitor of metalloproteinase-1 but not metalloproteinase-3 in atopic dermatitis. AB - Matrix metalloproteinases and their specific inhibitors, tissue inhibitors of metalloproteinases (TIMPs), contribute to inflammation-induced tissue destruction and subsequent remodeling for maintenance of tissue homeostasis. Since the production of these enzymes and their inhibitors is regulated by mediators such as proinflammatory cytokines and growth factors, elevated levels of serum TIMPs and/or MMPs have been documented in patients with several inflammatory disorders. In this study, we examined the role of TIMPs and MMPs in the pathogenesis of atopic dermatitis (AD) by evaluating the serum levels of TIMP-1 and MMP-3 in 40 patients with AD and 20 control subjects by ELISA. The serum TIMP-1 levels were significantly higher in AD patients in exacerbation status than in nonatopic subjects, whereas serum MMP-3 levels were not significantly different between them. As a result, AD patients revealed significantly elevated TIMP-1/MMP-3 ratios. The levels of serum TIMP-1 were significantly reduced in AD patients following conventional treatments. Significantly higher values of peripheral eosinophil counts, serum levels of IgE and lactate dehydrogenase, eruption score, and eruption area were noted in the AD patients with elevated TIMP-1 levels when compared with those with normal values. Moreover, the points of chronic eruptions such as lichenification and prurigo were significantly higher in the patients with elevated TIMP-1 levels than those with normal TIMP-1, while those of acute lesions such as oozy/microvesicles and oedema were not different between these groups. Serum TIMP-1 level may be a useful marker to estimate the long-term disease activity of AD. PMID- 11876752 TI - Anti-platelet antibodies associated with the Canale-Smith syndrome bind to the same platelet glycoprotein complexes as those of idiopathic thrombocytopenic purpura patients. AB - The Canale-Smith syndrome (CSS) is an inherited disease characterized by massive lymphadenopathy, hepatosplenomegaly and systemic autoimmunity to erythrocytes and platelets. Idiopathic thrombocytopenic purpura (ITP) is an autoimmune disease in which approximately 60-80% of patients have anti-platelet antibodies directed against specific platelet glycoprotein complexes (GPCs) located on their membrane: GP IIb/IIIa, GPIb/IX, and GPIa/IIa. Almost all (95-100%) of the antibody-positive patients have antibodies directed against GPIIb/IIIa alone, or in combination with other glycoprotein targets. Our objective was to determine the specificities of the anti-platelet antibodies in CSS patients. The detection of anti-platelet antibodies was performed using a commercially available ELISA, the Pak-AUTO (GTI, Brookfield, WI), in which highly purified GPIIb/IIIa, GPIb/IX, and GPIa/IIa are immobilized on microtitre plates, incubated with serum or plasma, and subsequently developed with an antihuman polyclonal immunoglobulin. Of 14 CSS patients tested, 11 (79%) had anti-platelet antibodies in their serum directed toward at least one of the three major GPC, nine (82%) of which were against GPIIb/IIIa alone or in combination. Antibodies detected in the sera of ITP patients had similar specificities. No such antibodies were detected in samples from 25 consecutive normal controls. These results demonstrate that a genetically defined defect in lymphocyte apoptosis results in a humoral autoimmune response with anti-platelet specificities very similar to the common idiopathic form of autoimmune thrombocytopenia. PMID- 11876753 TI - Autoantibodies against cytochrome P450s in sera of children treated with immunosuppressive drugs. AB - Treatment with the immunosuppressive drugs cyclosporin and tacrolimus, the mainstays of anti-graft rejection and autoimmune disease therapy, is limited by their hepato- and nephrotoxicity. The metabolic conversion of these compounds to more easily excretable products is catalysed mainly by hepatic cytochrome P4503A4 (CYP3A4) but also involves extrahepatic CYP3A5 and other P450 forms. We set out to study whether or not exposure to cyclosporin and FK506 in children undergoing organ transplantation leads to formation of autoantibodies against P450s. Immunoblotting analysis revealed anti-CYP reactivity in 16% of children on CyA for anti-graft rejection or treatment of nephrosis (n = 67), 31% of kidney transplant patients switched from CyA to FK506 (n = 16), and 21% of kidney and or liver transplant patients on FK506 (n = 14). In contrast, the frequency of reactive immunoblots was only 8.5% among the normal paediatric controls (n = 25) and 7% among adult kidney transplant patients on CyA or FK506 (n = 30). The CYP2C9+ sera were able to immunoprecipitate in vitro translated CYP2C9 and the immunoblot reactivity showed striking correlation to peaks in the age at onset of drug exposure. Sera were isoform selective as evidenced from Western blotting using human liver microsomes and heterologously expressed human P450s. These findings suggest that anti-cytochrome P450 autoantibodies, identified on the basis of their specific binding in immunoblots, are significantly increased among children on immunosuppressive drugs and in some cases are associated with drug toxicity and organ rejection. PMID- 11876754 TI - Local immunostimulation induced by intravesical administration of autologous interferon-gamma-activated macrophages in patients with superficial bladder cancer. AB - We conducted a phase I/II clinical trial of the safety and efficacy of intravesical administration of autologous IFN-gamma-activated macrophages (MAK) in patients with superficial bladder cancer. Monocyte-derived MAK cells were prepared in vitro and patients received six instillations of 1.4 x 10(8) to 2.5 x 10(8) cells, once a week, for five consecutive weeks. Treatment was well tolerated, with seven grade 1 and five Grade 2 protocol-related adverse effects. Nine out of 17 included patients had no recurrences during the year following the first instillation of MAK. The aim of the present study was to search for immune parameters related to local immunostimulation induced by MAK. Monitoring of the patients showed that urinary IL-8, GM-CSF and, to a lesser extent, IL-18 were increased following MAK instillations, with inter-individual differences. The urinary IL-8 level was about 10-fold higher than that observed for other cytokines, and its biological activity was reflected by a concomitant increase of urinary elastase, indicating neutrophil activation and degranulation. We also showed that nine out of 12 patients investigated presented an increase of urinary neopterin, a marker of IFN-gamma-activated macrophages, 7 days after MAK instillation, while serum neopterin levels were almost stable. These results are in line with persistence of activated macrophages in the bladder wall after infusions. Moreover, there was evidence of macrophages in urine smears 2 months after the sixth MAK instillation, and the score of macrophages correlated with the quantity of neutrophils in the urine. Overall, this study provides evidence of a local immunostimulation induced by this novel and safe immunotherapeutic approach of MAK instillations in patients with superficial bladder cancer. PMID- 11876755 TI - Complement activation in infective endocarditis: correlation with extracardiac manifestations and prognosis. AB - In an infectious process complement activation is necessary for a proper immune and inflammatory response, but when exacerbated may cause tissue injuries. In infective endocarditis (IE) patients tend to develop high titres of circulating immune complexes (CIC) that activate complement. The aim of this study was to evaluate for the first time complement activation in IE for possible correlation with extracardiac manifestations and clinical prognosis. Twenty patients with IE, 14 healthy controls and 15 patients presenting mitral and aortic valve lesions (with no signs of either infection or other associated diseases), were studied. Plasma levels of C3adesArg, SC5b-9, C1rs-C1Inh and C3b(Bb)P were determined by ELISA and C3d by double decker immunoelectrophoresis. C3 and C4 levels were assayed by turbidimetry and CIC by ELISA. Elevation of plasma levels of all complement activation products, with the exception of C3b(Bb)P, indicated a significant classical pathway activation in IE patients when compared to controls (C3d: P < 0.00004; C3adesArg: P < 0.03, SC5b-9: P < 0.01, C1rs-C1Inh: P < 0.00007). CIC levels were significantly increased (P < 0.005) and C3 reduced in IE patients (P < 0.05). Elevated C3d (P < 0.02) and C3adesArg (P < 0.03) levels were associated with pulmonary manifestations. In addition, C3d was significantly elevated in the patients who died when compared to those who had a good recovery (P < 0.02). Our data demonstrate the activation of the complement classical pathway, most probably mediated by CIC, in IE and suggests C3d and C3adesArg as possible markers for extracardiac lesion and severity of the disease. PMID- 11876756 TI - Smoking and disease severity are independent determinants of serum adhesion molecule levels in Graves' ophthalmopathy. AB - Adhesion molecules play a key role in autoimmune disorders, and serum concentrations of soluble adhesion molecules are increased in Graves' ophthalmopathy (GO). Whether this is due to the strong association with smoking is unknown. It is also not known if the severity or activity of GO determine the serum levels of adhesion molecules. We measured serum concentrations of sICAM-1, sVCAM-1 and sELAM-1 in 62 euthyroid Graves' patients with untreated GO, in 62 healthy controls matched for sex, age and smoking habits, and in 26 euthyroid Graves' patients without GO. GO severity was assessed by the Total Eye Score and the activity by the Clinical Activity Score. Adhesion molecules were measured by highly sensitive ELISAs. GO patients had higher levels than controls (median values in ng/ml with range): sICAM-1 300 [171--575] versus 244 [119--674], P < 0.001; sVCAM-1 457 [317--1060] versus 410 [238--562], P < 0.001; and sELAM-1 61 [19--174] versus 53 [23--118], P = 0.021. Euthyroid Graves' disease patients without GO had levels similar to controls: sICAM-1 273 138--453), sVCAM-1 386 [260--1041] and sELAM-1 46 [22--118]. Smoking had an independent effect and was associated with higher levels of sICAM-1 and lower levels of sVCAM-1 in both GO patients and controls; sELAM-1 levels were comparable. In the 62 GO patients, sICAM-1 correlated significantly with severity of eye disease (r = 0.40, P = 0.002). No correlation was found with the duration of GO, the Clinical Activity Score or TBII levels. Multivariate analysis of all 150 subjects showed that the presence of GO and smoking are independent determinants of sICAM-1 and sVCAM-1 concentrations. In GO patients, the Total Eye Score was a stronger determinant than smoking. It is concluded that (i) smoking is associated with increased sICAM 1 and decreased sVCAM-1 levels; (ii) independent from smoking, euthyroid GO patients have higher levels of sICAM-1, sVCAM-1 and sELAM-1 than patients with euthyroid Graves' disease or healthy controls; (iii) the major determinant of sICAM-1 in GO patients is the severity of their eye disease. PMID- 11876757 TI - Expression and functional activity of CXCR-4 and CCR-5 chemokine receptors in human thymocytes. AB - In this paper we addressed the expression of the HIV co-receptors CXCR-4 and CCR 5 in human thymocytes by phenotypic, molecular and functional approaches. Cytofluorimetric analysis disclosed that CXCR-4 was constitutively expressed by freshly isolated thymocytes (~10 000 molecules/cell in about 30% of thymocytes); the receptor was endowed with functional activity, as it mediated polarization, migration and intracellular Ca2+ increase in response to its ligand, SDF-1. On the contrary, CCR-5 expression in freshly isolated thymocytes was significantly lower (<4000 molecules/cell in less than 5% of the cells), and no functional response to CCR-5 agonists could be documented. Northern blot analysis of freshly isolated thymocytes showed high CXCR-4 mRNA levels, whereas the message for CCR-5 was barely detectable. On the other hand, a modest increase in the expression of CCR-5 was associated with in vitro thymocyte stimulation, and CCR-5 density at the cell surface attained CXCR-4 figures in most cases. None the less, no functional response to CCR-5 agonists could be documented in in vitro stimulated thymocytes. In vitro infection of thymocytes by CAT-expressing recombinant HIV bearing the envelope glycoproteins from different isolates showed that T-tropic strains, which use CXCR-4 as a co-receptor, were more efficient in infecting thymocytes than M-tropic strains, which preferentially use CCR-5. Altogether, these data indicate that expression of the major co-receptors involved in infection by M-tropic HIV strains is very poor in human thymocytes, and would suggest that thymocyte infection by M-tropic HIV strains may be a rare event in vivo. PMID- 11876758 TI - Interleukin-1 receptor antagonist gene polymorphism and mortality in patients with severe sepsis. AB - This study aims to determine the influence of the polymorphism within the intron 2 of the interleukin-1 receptor antagonist gene (IL-1RN*) on the outcome of severe sepsis, and to assess its functional significance by correlating this polymorphism with the total production of interleukin-1 receptor antagonist (IL 1Ra) protein determined in stimulated peripheral blood mononuclear cells (PBMC). A group of 78 patients with severe sepsis (51 survivors and 27 nonsurvivors) was compared with a healthy control group of 130 blood donors, and 56 patients with uncomplicated pneumonia. We found a significant association between IL-1RN* polymorphism and survival. Thus, after adjusting for age and APACHE II score, multiple logistic regression analysis showed that patients homozygotes for the allele *2 had a 6.47-fold increased risk of death (95% CI 1.01--41.47, P = 0.04). Besides, compared with patients homozygous or heterozygous for the allele *1, IL 1RN*2 homozygotes produced significantly lower levels of IL-1Ra from their PBMC. Our results suggest that insufficient production of this cytokine might contribute, among other factors, to the higher mortality rate found in severe sepsis patients with the IL-1RN*2 homozygous genotype. PMID- 11876759 TI - Monocyte intracellular cytokine production during human endotoxaemia with or without a second in vitro LPS challenge: effect of RWJ-67657, a p38 MAP-kinase inhibitor, on LPS-hyporesponsiveness. AB - In the present study, we investigated the effect of RWJ-67657, a p38 MAP kinase inhibitor, upon in vivo LPS-induced monocyte cytokine production and upon monocyte LPS-hyporesponsiveness. Thirty minutes before a single injection of LPS (4 ng/kg BW), healthy male volunteers received a single oral dose of RWJ-67657 at increasing dosages (0-1400 mg). Blood samples (pre-medication, 3, 6 and 24 h after LPS) were immediately incubated with LPS (reflecting LPS hyporesponsiveness) or without LPS (reflecting in vivo monocyte stimulation) for 4 h at 37 degrees C. Following red blood cells lysis and white blood cell permeabilization, cells were labelled with alpha-CD14-FITC and alpha-IL-1beta, alpha-IL-12 or alpha-TNFalpha (PE-labelled), fixed, and analysed using flow cytometry. In vivo LPS injection resulted in an increased percentage of circulating monocytes producing IL-1beta, TNFalpha and IL-12 only at 3 h after the LPS injection. This was dose-dependently inhibited by RWJ-67657 treatment. LPS-hyporesponsiveness to in vitro LPS treatment was most prominent at 3 and 6 h after the in vivo LPS injection; compared with pre-medication monocytes, at these intervals a reduced percentage of monocytes produced IL-1beta, TNFalpha or IL-12 after the in vitro LPS stimulus. At t = 6 h, this LPS-hyporesponsiveness could dose-dependently be inhibited by RWJ-67657 treatment of the volunteers. We therefore conclude that p38 MAP kinase inhibition with RWJ-67657 inhibited monocyte production of cytokines following in vivo LPS injection. Treatment with RWJ-67657 also reversed the LPS-hyporesponsiveness. Whether this result can be extended to the clinical situation remains to be elucidated. Patients with sepsis or an otherwise high risk for multi-organ failure are potential study groups. PMID- 11876760 TI - Age-dependent preference in human antibody responses to Streptococcus pneumoniae polypeptide antigens. AB - Vulnerability to Streptococcus pneumoniae is most pronounced in children. The microbial virulence factors and the features of the host immune response contributing to this phenomenon are not completely understood. In the current study, the humoral immune response to separated Strep. pneumoniae surface proteins and the ability to interfere with Strep. pneumoniae adhesion to cultured epithelial cells were analysed in adults and in children. Sera collected from healthy adults recognized Strep. pneumoniae separated lectin and nonlectin surface proteins in Western blot analysis and inhibited on average 80% of Strep. pneumoniae adhesion to epithelial cells in a concentration-dependent manner. However, sera longitudinally collected from healthy children attending day care centres from 18 months of age and over the course of the following 2 years revealed: (a) development of antibodies to previously unrecognized Strep. pneumoniae surface proteins with age; (b) a quantitative increase in antibody responses, measured by densitometry, towards separated Strep. pneumoniae surface proteins with age; and (c) inhibition of Strep. pneumoniae adhesion to epithelial cells, which was 50% on average at 18 months of age, increased significantly to an average level of 80% inhibition at 42 months of age equalling adult sera inhibitory values. The results obtained in the current study, from the longitudinally collected sera from healthy children with documented repeated Strep. pneumoniae colonization, show that repeated exposures are insufficient to elicit an immune response to Strep. pneumoniae proteins at 18 months of age. This inability to recognize Strep. pneumoniae surface proteins may stem from the inefficiency of T-cell-dependent B-cell responses at this age and/or from the low immunogenicity of the proteins. PMID- 11876762 TI - IL-15 in human visceral leishmaniasis caused by Leishmania infantum. AB - Interleukin (IL)-15 is a recently discovered cytokine with the ability to stimulate the proliferation activity of Th1 and/or Th2 lymphocytes. Here, we investigated the involvement of IL-15 in the immune response to Leishmania infantum infection by studying patients with visceral leishmaniasis (VL). We found that IL-15 is produced by leishmanial antigen (LAg)-stimulated peripheral blood mononuclear cells (PBMC) from active VL patients at a significantly higher level than those produced by cells from healed VL subjects or healthy controls. A significant increase in IL-15 serum blood levels was also observed in acute VL patients compared with healed ones. Furthermore, recombinant IL-15 had an appreciable effect in vitro in reducing IL-4 and increasing the production of IL 12 in response to LAg, but it was ineffective in altering the production of interferon-gamma (IFN-gamma). The production of endogenous IL-15 in acute VL patients appeared to be insufficient to activate both IFN-gamma and IL-12, as attested by the absence of modification of these two cytokines by neutralization experiments in the presence of anti-IL-15 monoclonal antibodies (MoAB). On the contrary, the neutralization of IL-15 increased IL-4 production. Together, these results indicate that endogenous IL-15 plays a role in the suppression of Th2 type cytokines, even though it does not enhance the production of Th1 cytokines in acute VL patients. Since IL-15, in the presence of anti-IL-4 MoAb, caused a further increase in IL-12 production and led to a significant production of IFN gamma, one of its indirect effects on Th1 cell activation could be due to the latter's effect on Th2 cytokines such as IL-4. Therefore, our observations indicate that there is a potential for IL-15 to augment the T-cell response to human intracellular pathogens. PMID- 11876761 TI - Reduced efficacy of treatment of strongyloidiasis in HTLV-I carriers related to enhanced expression of IFN-gamma and TGF-beta1. AB - Strongyloidiasis, a human intestinal infection caused by Strongyloides stercoralis (S. stercoralis), is difficult to cure with drugs. In particular, a decrease of the efficacy of treatment has been reported in patients dually infected with S. stercoralis and human T-cell leukaemia virus type I (HTLV-I), both of which are endemic in Okinawa, Japan. However, the factors influencing this resistance remain unclear. In the present study, patients infected with S. stercoralis, with or without HTLV-I infection, were treated with albendazole, followed up for one year and separated into two groups, cured and non-cured. The cure rate of S. stercoralis was lower in HTLV-I carriers (P < 0.05). Serum levels of S. stercoralis-specific IgA, IgE, IgG, IgG1 and IgG4 antibodies were estimated, and a decrease of IgE (P < 0.05) and an increase of IgG4 (P < 0.05) were observed in the non-cured group, especially in HTLV-I carriers. RT-PCR of cytokines using peripheral blood mononuclear cells revealed that S. stercoralis patients with HTLV-I showed a high frequency of expression of IFN-gamma and TGF beta1, whereas those without HTLV-I showed no expression of these cytokines. IFN gamma- and TGF-beta1-positive HTLV-I carriers showed a decrease of IgE (P < 0.05), an increase of IgG4 (P < 0.01) and a lower cure rate (P < 0.01) compared with those who were negative for both cytokines. These results suggest that persistent infection with HTLV-I affected S. stercoralis-specific immunity and reduced therapeutic efficacy. PMID- 11876763 TI - Characterization of Herpesvirus saimiri-transformed T lymphocytes from common variable immunodeficiency patients. AB - Common variable immunodeficiency (CVID) is a very frequent but heterogeneous syndrome of antibody formation. The primary defect remains unknown, but many reports describe peripheral blood T lymphocyte dysfunctions in a substantial proportion of CVID patients, which may impair T--B cell collaboration. In order to investigate whether such putative defects were intrinsic to T cells or, rather, secondary to quantitative differences in T cell subset distribution, or to other described disorders, we have used Herpesvirus saimiri (HVS) for the targeted transformation of CVID CD4+ and CD8+ T cells and subsequent functional evaluation by flow cytometry of their capacity to generate cell surface (CD154, CD69) or soluble (IL-2, TNF-alpha, IFN-gamma) help after CD3 engagement. Unexpectedly, the results showed that 40 different CVID blood samples exposed to HVS gave rise with a significantly increased frequency to transformed CD4+ T cell lines, compared to 40 age-matched controls (27% versus 3%, P < or = 0.00002) suggesting the existence of a CVID-specific signalling difference which affects CD4+ cell transformation efficiency. The functional analysis of 10 CD4+ and 15 CD8+ pure transformed T cell lines from CVID patients did not reveal any statistically significant difference as compared to controls. However, half of the CD4+ transformed cell lines showed CD154 (but not CD69) induction (mean value of 46.8%) under the lower limit of the normal controls (mean value of 82.4%, P < or = 0.0001). Exactly the same five cell lines showed, in addition, a significantly low induction of IL-2 (P < or = 0.04), but not of TNF-alpha or IFN gamma. None of these differences were observed in the remaining CD4+ cell lines or in any of the transformed CD8+ cell lines. We conclude that certain CVID patients show selective and intrinsic impairments for the generation of cell surface and soluble help by CD4+ T cells, which may be relevant for B lymphocyte function. The transformed T cell lines will be useful to establish the biochemical mechanisms responsible for the described impairments. PMID- 11876764 TI - A type 2 response in lipopolysaccharide (LPS)-stimulated whole blood cell cultures from periodontitis patients. AB - It is acknowledged that periodontitis results from the interaction of the host immune response with bacteria accumulating on the tooth surfaces. Although bacteria are essential, they are insufficient to cause the disease. Despite this knowledge it remains unclear why certain individuals are more susceptible to periodontitis than others. Therefore the present study investigated whether differences exist in the actual immune response between periodontitis patients and controls after stimulation of peripheral blood cells. Whole blood cell cultures (WBCC) were stimulated with LPS from Escherichia coli during 18 h and the release of prostaglandin E2 (PGE2), IL-1beta, IL-6, IL-8, IL-10, IL-12p40, IL 12p70 and tumour necrosis factor-alpha (TNF-alpha) was measured. The levels of PGE2 were two-fold higher in the WBCC from periodontitis patients than from controls. In contrast, the levels of IL-12p70 in WBCC from patients were two-fold lower. Furthermore, WBCC from patients secreted lower levels of IL-1beta and higher levels of IL-8 when compared with WBCC from controls. No differences were observed with respect to IL-6, IL-10, IL-12p40 and TNF-alpha production. It is known from the literature that LPS-stimulated WBCC reflect specifically the behaviour of the monocytes and that monocytes are peripheral precursors of antigen-presenting cells (APC). Therefore it is concluded that the monocytes in the present WBCC from periodontitis patients are responsible for the higher levels of PGE2 and lower levels of IL-12p70. Since it is has been shown that APC derived IL-12p70 induces type (Th1) cells that promote cellular immunity, while APC-derived PGE2 induces type 2-helper (Th2) cells that promote humoral immunity, it is postulated that APC from periodontitis patients may have a bias in directing Th2 responses and thereby promoting the humoral immunity in periodontitis. PMID- 11876765 TI - Analysis of the B cell repertoire against autoantigens in patients with giant cell arteritis and polymyalgia rheumatica. AB - The analysis of the antibody repertoire of patients with giant cell arteritis (GCA) and polymyalgia rheumatica (PMR) might identify target antigens of the autoimmune response with potential relevance to our understanding of the pathogenesis of the disease and to the development of serodiagnostic tests. To detect such antigens, we screened a cDNA library derived from normal human testis for antigens reacting with IgG antibodies in the 1 : 250 diluted sera of three patients with untreated GCA using SEREX, the serological identification of antigens by recombinant cDNA expression cloning. Of 100 000 clones screened with each serum, six, 28 and six clones, respectively, were positive, representing a total of 33 different antigens. Most of the antigens reacted only with the serum used for identification and/or at a similar frequency with normal control sera. However, lamin C and the nuclear antigen of 14 kD reacted specifically with 32% of GCA/PMR, but with none of the control sera, while human cytokeratin 15, mitochondrial cytochrome oxidase subunit II, and a new gene product were detected preferentially, but not exclusively by sera from GCA/PMR patients. We conclude that patients with GCA/PMR develop antibodies against a broad spectrum of human autoantigens. Antibodies against human lamin C, the nuclear autoantigen of 14 kD as well as human cytokeratin 15, mitochondrial cytochrome oxidase subunit II and the product of a new gene should be investigated further to determine their value as tools for the diagnosis and/or the definition of clinical subgroups of patients with GCA/PMR. PMID- 11876766 TI - CD40 on salivary gland epithelial cells: high constitutive expression by cultured cells from Sjogren's syndrome patients indicating their intrinsic activation. AB - CD40 has been identified in an expanding list of haematopoietic and non haematopoietic cells and has received an increased interest based on its role in a variety of cell-mediated responses and its potential to participate in the pathogenesis of chronic inflammatory disorders. Sjogren's syndrome (SS) is an autoimmune exocrinopathy, which is characterized by chronic lymphocytic infiltration of exocrine glands and aberrant activation of epithelial tissues. We studied the expression of CD40 protein in cultured non-neoplastic salivary gland epithelial cell (SGEC) lines as well as in minor SG biopsies obtained from 17 SS patients and 12 controls. Immunocytochemical and flow cytometric analyses had revealed the occurrence of constitutively expressed CD40 molecules on the surface of long-term cultured SGEC lines, which could be further induced by interferon gamma (IFN-gamma) and IL-1beta cytokines, but not tumour necrosis factor-alpha (TNF-alpha), IL-4, IL-6, granulocyte-macrophage colony-stimulating factor (GM CSF) or IFN-alpha. Triggering of SGEC through CD40 enhanced the surface expression of the adhesion molecule intercellular adhesion molecule-1 (ICAM 1)/CD54, but not MHC class I and class II (HLA-DR) molecules. Spontaneous CD40 expression was significantly higher in SGEC lines derived from SS patients, compared with controls (P < 0.001), which is suggestive of their intrinsically activated status. In SG biopsies, CD40 was constitutively expressed by lymphocytes, ductal epithelial cells and endothelial cells but not by other glandular cell types, such as acinar cells, myoepithelial cells and fibroblasts. In addition, CD40L staining was also detected in 30--50% of the infiltrating lymphocytes in the biopsies of SS patients. Our findings indicate the immunoregulatory potential of SGEC and lend further support to a model of intrinsic activation in salivary epithelia in SS, whereby these cells actively participate in the induction and maintenance of lymphocytic infiltrates of patients. PMID- 11876768 TI - A single olfactory receptor specifically binds a set of odorant molecules. AB - The sense of smell is mediated by the initiation of action potential in olfactory sensory neurons during odor stimulation. However, little is known about odorant olfactory receptor (OR) recognition mechanisms. In the present work, we identified the structural motifs of odorant molecules required to activate mouse OR912-93 by detection of the odorant response using calcium measurement in cells transfected with OR and G(alpha)q and G(alpha)15 proteins. The use of sets of odorants led to the identification of ketones with an aliphatic carbon chain length >or= four carbon atoms and a carbonyl group preferentially located in position C2 or C3. The threshold of detection of these odorants is as low as 10( 6)-10(-8)m. No other odorant ligand, out of 70 representatives of the odorant world, was active. The human ortholog of OR912-93 is not functional, suggesting that apart from a stop-mutation located at the 5'-end that was corrected in the construct, it incurred other deleterious mutations during evolution. PMID- 11876769 TI - NMDA antagonists exacerbate neuronal death caused by proteasome inhibition in cultured cortical and striatal neurons. AB - The proteasome is involved in multiple cellular processes including control of the cell cycle, apoptosis and intracellular signalling; loss of proteasome function has been postulated to participate in the pathogenesis of triplet repeat diseases. We examined the vulnerability of central neurons to proteasome inhibition and tested the ability of anti-excitotoxic and anti-apoptotic treatments to attenuate proteasome inhibition-induced neuronal death. Exposure of murine neocortical cultures to proteasome inhibitors (0.1-10 microm clasto lactacystin beta-lactone or MG-132) for 48 h resulted in widespread neuronal death associated with a reduction in intracellular free calcium; higher inhibitor concentrations killed astrocytes. Cultured striatal neurons were more vulnerable than cortical neurons. Within each population, the NADPH diaphorase-positive neuronal subpopulation was more vulnerable than the general neuronal population. Enhancing calcium entry with S(-)BayK8644 or kainate, or blocking apoptosis with cycloheximide, actinomycin D or Z-VAD.FMK attenuated neuronal death, whereas, reducing calcium entry with NMDA antagonists or R(+)BayK8644 potentiated neuronal death. These findings suggest that proteasome inhibition can induce selective neuronal apoptosis associated with intracellular calcium starvation, and point to manipulation of intracellular calcium as a specific therapeutic strategy. In particular, concern is raised that glutamate receptor antagonists might exacerbate, rather than attenuate, proteasome inhibition-induced neuronal death. PMID- 11876767 TI - Activity of matrix metalloproteinase-9 is elevated in sera of patients with systemic lupus erythematosus. AB - Systemic lupus erythematosus (SLE) is an autoimmune disease characterized by the increased production of autoantibodies and by systemic clinical manifestations and damage to multiple organs. The aim of the present study was to analyse matrix metalloproteinase (MMP)-9 activity in sera of patients with active and inactive SLE in order to evaluate its role in the pathogenesis and course of the disease, as well as its diagnostic value. We measured activity levels of MMP-9 and MMP-2, using both gel zymography and activity assay kits, in sera of 40 SLE patients and of 25 healthy controls. We found that MMP-9 activity, but not MMP-2 activity, is significantly elevated in the sera of SLE patients compared with sera samples of healthy controls. High activity levels of MMP-9 were determined in sera of 68% of the SLE patients. Elevated levels of MMP-9 were correlated with the presence of discoid rash, Raynaud phenomenon, pneumonitis, mucosal ulcers and anti phospholipid antibodies. Changes in activity levels of MMP-9, but not of MMP-2, were observed in sera of the same patient at different periods of the disease course. High levels of MMP-9 did not correlate with disease activity index (SLEDAI, BILAG) in female patients, but correlated with SLE activity in the group of male patients. The results of the present study suggest that MMP-9 plays a role in the pathogenesis of SLE. PMID- 11876770 TI - Modulation by 5-hydroxytryptamine of nicotinic acetylcholine responses recorded from an identified cockroach (Periplaneta americana) motoneuron. AB - Recordings from the soma of the cockroach (Periplaneta americana) fast coxal depressor motoneuron (Df) were made while acetylcholine (ACh) was regularly pressure-applied locally from a micropipette. The modulatory effects upon these nicotinic ACh responses of bath-applied 5-hydroxytryptamine (5-HT, serotonin), dopamine and octopamine were investigated under either current-clamp or voltage clamp conditions. The biogenic amines reversibly suppressed, but never totally abolished, ACh responses, 5-HT being the most potent, with a threshold near 10( 6) m (EC50 = 5 x 10(-5) m). Occlusion experiments indicate that the amines share a common mechanism at the level of either receptors or second messenger pathways. The amines also modulated responses to nicotine or carbachol (each of which resists hydrolysis by acetylcholinesterases), indicating that the amines did not act by accelerating ACh degradation. Pharmacological antagonists were used in an attempt to characterize the receptor responsible for amine-mediated modulation. Although a number of antagonists mimicked the action of amines rather than producing blockade, the antagonistic actions of LSD and RS23597 pointed strongly to a receptor-mediated mechanism, but did not allow receptor identification. The magnitude of the modulatory effect of 5-HT was significantly reduced by intracellular guanosine-5'-O-(2-thiodiphosphate) (GDP-beta-S), indicating involvement of a G-protein. Intracellular injection of the calcium chelator BAPTA did not block the modulatory effect of 5-HT, showing that the amines do not operate through the calcium-dependent pathway by which muscarinic receptors act on nicotinic currents. The adenylate cyclase inhibitor dideoxyadenosine (DDA), on the other hand, did attenuate the action of 5-HT, suggesting involvement of cyclic AMP. PMID- 11876771 TI - Regulation of acetylcholine release by muscarinic receptors at the mouse neuromuscular junction depends on the activity of acetylcholinesterase. AB - Muscarinic acetylcholine receptors (mAChRs) play an important role in regulating the release of acetylcholine (ACh) in various tissues. We used subtype-specific antibodies and a fluorescent-labelled muscarinic toxin to demonstrate that mammalian neuromuscular junction expresses mAChR subtypes M1 to M4, and that localization of all subtypes is highly restricted to the innervated part of the muscle. To elucidate the roles of the mAChR subtypes regulating ACh release, we measured the mean quantal content of endplate potentials in isolated mouse phrenic--hemidiaphragm preparations in which release was reduced by a low Ca2+/high Mg2+ medium. Muscarine decreased evoked ACh release in normal junctions but, depending on the concentration, reduced or increased transmitter release in collagen Q-deficient junctions completely lacking acetylcholinesterase (AChE). Both effects were also seen in normal junctions when AChE was inhibited by various doses of fasciculin-2. Block of mAChRs by atropine had no effect on evoked release at normal junctions, but decreased release at junctions lacking AChE. The muscarine-elicited depression of ACh release in normal junctions was completely abolished by pertussis toxin or methoctramine pretreatment, but was not affected by muscarinic toxin MT-3, thus indicating the involvement of the M2 mAChR. The muscarine-induced increase of ACh release in AChE-deficient junctions was not affected by pertussis toxin, but was completely blocked by MT-7, a specific M1 mAChR antagonist. Our results show that the M1 and M2 mAChRs have opposite presynaptic functions in modulating quantal ACh release, and that regulation of release by the two receptor subtypes depends on the functional state of AChE at the neuromuscular junction. PMID- 11876772 TI - Differential subcellular localization of the 5-HT3-As receptor subunit in the rat central nervous system. AB - Following the cloning and sequencing of the A subunit of the 5-HT3 receptor, two alternatively spliced isoforms, 5-HT3-AS and 5-HT3-AL, have been identified. In order to analyse the distribution of the receptor, a polyclonal antibody has been produced against the short form which is the most abundant in the central nervous system [Doucet et al. (2000) Neuroscience 95, 881-892]. As expected from the recognition of functional 5-HT3 receptors, immunostaining by this anti-5-HT3-R-AS antibody matched the distribution of the high-affinity 5-HT3 binding sites in the rat brain and spinal cord. 5-HT3-AS-like immunoreactivity was detected at low levels in the limbic system, particularly in the amygdala and the hippocampus, and in the frontal, piriform and entorhinal cortices. High levels of immunoreactivity were found in the brainstem, mainly in the nucleus tractus solitarius and the nucleus of the spinal tract of the trigeminal nerve, and in the dorsal horn of the spinal cord. At the ultrastructural level, immunostaining was generally found associated with axons and nerve terminals (70-80%) except in the hippocampus, where labelled dendrites were more abundant (56%). This preferential localization on nerve endings is consistent with the well-documented physiological role of 5-HT3 receptors in the control of neurotransmitter release. However, the different distribution in the hippocampus raises the question of whether differential addressing mechanisms exist for preferentially targeting 5 HT3 receptors to postsynaptic dendritic sites as compared to presynaptic nerve endings, depending on the nature of the neurons bearing these receptors. PMID- 11876773 TI - Nitric oxide-dependent damage to neuronal mitochondria involves the NMDA receptor. AB - Cytokine-stimulated astrocytes produce nitric oxide, which can inhibit components of the mitochondrial respiratory chain. We have previously demonstrated that prolonged exposure (48 h) to rat astrocytic nitric oxide damages complexes II- III and IV of neighbouring rat neurons in coculture, resulting in neuronal death. Expanding on these observations, we have now shown that the NMDA receptor antagonist, MK-801, prevents this damage, suggesting involvement of glutamate. We postulate that astrocyte-derived nitric oxide stimulates release of neuronal glutamate. Indeed we demonstrate that neurons incubated with nitric oxide generating astrocytes display enhanced glutamate release. Furthermore, direct exposure to the nitric oxide donor, DETA-NONOate resulted in a loss of activity of all the neuronal mitochondrial complexes, which was again prevented by MK-801. Thus, nitric oxide, generated by both cytokine-stimulated astrocytes and by a nitric oxide donor, causes activation of the NMDA receptor leading to damage to the neuronal mitochondrial respiratory chain. Glutamate exposure is known to damage the neuronal mitochondrial respiratory chain via neuronal nitric oxide synthase. Therefore, we propose that astrocyte-derived nitric oxide is capable of eliciting neuronal glutamate release, which in turn activates the neuronal NMDA receptor and stimulates further formation of reactive nitrogen species via neuronal nitric oxide synthases, leading to mitochondrial damage and neuronal death. Our findings support the hypothesis that glutamate, reactive nitrogen species and mitochondrial dysfunction may have a role in the neurodegenerative process. PMID- 11876775 TI - Learning the invariance properties of complex cells from their responses to natural stimuli. AB - Neurons in primary visual cortex are typically classified as either simple or complex. Whereas simple cells respond strongly to grating and bar stimuli displayed at a certain phase and visual field location, complex cell responses are insensitive to small translations of the stimulus within the receptive field [Hubel & Wiesel (1962) J. Physiol. (Lond.), 160, 106-154; Kjaer et al. (1997) J. Neurophysiol., 78, 3187-3197]. This constancy in the response to variations of the stimuli is commonly called invariance. Hubel and Wiesel's classical model of the primary visual cortex proposes a connectivity scheme which successfully describes simple and complex cell response properties. However, the question as to how this connectivity arises during normal development is left open. Based on their work and inspired by recent physiological findings we suggest a network model capable of learning from natural stimuli and developing receptive field properties which match those of cortical simple and complex cells. Stimuli are drawn from videos obtained by a camera mounted to a cat's head, so they should approximate the natural input to the cat's visual system. The network uses a competitive scheme to learn simple and complex cell response properties. Employing delayed signals to learn connections between simple and complex cells enables the model to utilize temporal properties of the input. We show that the temporal structure of the input gives rise to the emergence and refinement of complex cell receptive fields, whereas removing temporal continuity prevents this processes. This model lends a physiologically based explanation of the development of complex cell invariance response properties. PMID- 11876774 TI - Phosphorylation of GluR4 AMPA-type glutamate receptor subunit by protein kinase C in cultured retina amacrine neurons. AB - We have previously reported that the activity of alpha-amino-3-hydroxy-5-methyl-4 isoxazole propionate (AMPA) receptors is potentiated by protein kinase C (PKC) in cultured chick retina amacrine neurons, and that constitutive PKC activity is necessary for basal AMPA receptor activity (Carvalho et al., 1998). In this study, we evaluated the phosphorylation of the GluR4 subunit, which is very abundant in cultured amacrine neurons, to correlate it with the effects of PKC on AMPA receptor activity in these cells. 32P-labelling of GluR4 increased upon AMPA receptor stimulation or cell treatment with phorbol 12-myristate 13-acetate (PMA) before stimulating with kainate. By contrast, phosphorylation of GluR4 was not changed when PKC was inhibited by treating the cells with the selective PKC inhibitor GF 109203X before stimulation with kainate. We conclude that GluR4 is phosphorylated upon PKC activation and/or stimulation of AMPA receptors in cultured amacrine cells. Additionally, AMPA receptor activation with kainate in cultured chick amacrine cells leads to translocation of conventional and novel PKC isoforms to the cell membrane, suggesting that PKC could be activated upon AMPA receptor stimulation in these cells. PMID- 11876776 TI - Ultrastructural evaluation of calcitonin gene-related peptide immunoreactivity in the human cochlea and vestibular endorgans. AB - Calcitonin gene-related peptide (CGRP) is a neuropeptide widely distributed in the peripheral and central nervous system. Demonstrated in the efferent systems of the mammalian cochlea and vestibule, immunoreactive patterns of CGRP may vary by species. There is, however, no information in the literature investigating CGRP localization in the human cochlea. In the present study, the ultrastructural localization of CGRP immunoreactivity was evaluated in the human inner ear with immunoelectron microscopy. It was found that, in human cochlea, CGRP immunoreactivity was located in unmyelinated nerve fibres of the spiral lamina, inner spiral fibres beneath inner hair cells, tunnel spiral fibres, tunnel crossing fibres and outer radial fibres. In endorgans of human vestibule, CGRP immunoreactivity was located in vesiculated nerve fibres and bouton-type nerve terminals which were seen to contact afferent nerve chalices surrounding type I sensory cells and afferent nerve fibres, or to form an en passant contact with afferent dendrites. CGRP immunoreactivity appeared to be confined to efferent systems in all cases. This study presents evidence that CGRP could serve a role in neurotransmission or neuroregulation in both cochlear and vestibular efferent systems of human. PMID- 11876777 TI - Cortico-hippocampal APP and NGF levels are dynamically altered by cholinergic muscarinic antagonist or M1 agonist treatment in normal mice. AB - To determine whether altered cholinergic neurotransmission can modify the long term secretion of amyloid precursor protein (APP), endogenous levels of APP and nerve growth factor (NGF), we administered a selective M1 muscarinic receptor agonist (RS86) or the muscarinic antagonist, atropine, for 7 days in vivo into young adult mice (C57BL/6j). The levels of NGF and total APP in the hippocampus, frontal cortex, striatum, parietal cortex and cerebrospinal fluid (CSF) were examined by ELISA and Western blot. We found that this repeated i.m. administration of M1 receptor agonist resulted in decreased total APP levels in the hippocampus, frontal cortex and parietal cortex, and increased secreted alpha APPs levels in the CSF. M1 agonist treatment also resulted in decreased NGF levels in the hippocampus and CSF. These effects of the M1 muscarinic agonist could be blocked by atropine, which by itself elevated tissue levels of total APP. Interestingly, we found that the decrease of total APP in the hippocampus and striatum after M1 agonist treatment inversely correlated with the change in NGF levels. These data suggest that a sustained increased cholinergic, M1 mediated neurotransmission will enhance secretion of alpha-APPs in CSF and adaptively reduce the levels of total APP and NGF in the corticohippocampal regions of normal mice. The dynamic and adaptive regulation linking total APP and NGF levels in normal adult mice is relevant for understanding the pathophysiology of conditions with cholinergic and APP related pathologies, like Alzheimer's disease and Down's syndrome. PMID- 11876778 TI - Crossed unilateral lesions of temporal lobe structures and cholinergic cell bodies impair visual conditional and object discrimination learning in monkeys. AB - Monkeys with excitotoxic lesions of the CA1/subiculum region in the right hemisphere and with immunotoxic lesions of the cholinergic cells of the diagonal band in the left hemisphere were impaired on a visual conditional task. In this task, correct choice of one of two objects depends on which of two background fields both objects are presented against, irrespective of the spatial positions of the objects. They were not impaired on simple object or shape discrimination tasks. The pattern of impairments is the same as that seen after bilateral excitotoxic lesions of CA1/subiculum, implying that the diagonal band lesion disables the ipsilateral CA1/subiculum. It also argues that CA1/subiculum, sustained by its cholinergic input, is necessary for some forms of nonspatial conditional learning. Addition of an inferotemporal (IT) cortical ablation to the left hemisphere did not affect simple visual discrimination learning, although all the monkeys then failed to learn a new visual conditional task. This demonstrates that intact IT cortex in only one hemisphere is sufficient to sustain simple visual discrimination learning but implies that the cholinergic input and the inferotemporal cortical input to the hippocampus both contribute to visual conditional learning. The subsequent addition of an immunotoxic lesion of the basal nucleus of Meynert in the right hemisphere resulted in an additional impairment on a difficult shape discrimination. This argues that it is the cholinergic projection to the inferotemporal cortex, rather than to the rest of the cortex, which contributes to visual discrimination learning and memory. PMID- 11876779 TI - Novel expression of AMPA-receptor subunit GluR1 on mossy cells and CA3 pyramidal neurons in the human epileptogenic hippocampus. AB - Previous immunocytochemical investigations performed in our laboratory on the human hippocampus surgically resected for the treatment of mesial temporal lobe epilepsy (MTLE) have demonstrated an increased expression of the AMPA-receptor subunit GluR1 on neurons in the hilus and area CA3. Light microscopically, many of these neurons exhibited peculiar filamentous extensions and grape-like excrescences that protruded from their somata and proximal dendrites, suggesting that these neurons may be mossy cells and CA3 pyramidal neurons, respectively. The present electron microscopic study was carried out to further characterize these cells. The filamentous extensions were identified as dendrites from which spines often protruded, and the grape-like excrescences represented clusters of closely associated dendrites and spines. A variety of synapses were formed by the GluR1-positive profiles. These arrangements ranged from simple contacts between a single unlabelled axon terminal and a single labelled postsynaptic element, to complex contacts involving multiple unlabelled axon terminals and labelled postsynaptic elements. Many of the axon terminals involved in these arrangements were mossy fibre boutons. Thus, a large proportion of the GluR1-positive neurons were identified as hilar mossy cells and CA3 pyramidal neurons, cells hitherto thought to be absent or greatly reduced in the MTLE hippocampus. Taken together, these data suggest the presence of a highly efficient excitatory circuit involving AMPA receptors, mossy cells and CA3 pyramidal neurons in the sclerotic hippocampus. Such a circuit could be critically involved in the genesis and maintenance of temporal lobe epilepsy. PMID- 11876780 TI - Effects of water on cortical excitability in humans. AB - The effects of water on cortical excitability, measured using magnetoencephalographic recordings, were investigated in a sample of 19 healthy volunteers in a double-blind, placebo experiment comparing water with saline solution. Spontaneous magnetoencephalogram as well as auditory-evoked magnetic fields were recorded before and after the drinking of 750 mL water (9 subjects) or saline solution (10 subjects) and during and after hyperventilation following the drinking conditions. Hyperventilation was used to enhance the hypothesized synchronizing effect of water on spontaneous magnetoencephalographic activity. In addition, the magnetic fields were measured during a dichotic listening task under attended and unattended conditions. The prediction, that intake of water, because of induced cell swelling, will increase neuronal excitability and lead to an increased synchronization of the spontaneous magnetoencephalogram during hyperventilation was confirmed. Hyperventilation induced an increase of spectral power in all frequency bands particularly theta and delta power after water drinking. Furthermore, there was an increase of magnetic mismatch negativity (MMNm) amplitude in attended conditions and a simultaneous decrease in unattended conditions after water drinking. N1m (magnetic N1 wave) revealed significant changes during experimental conditions: increase after drinking and decrease after hyperventilation in both groups. MMNm for attended conditions showed a high positive correlation with osmolality changes (difference in the mol solute per kg water before and after drinking); N1m and PNm (magnetic processing negativity) as well as MMNm for unattended conditions showed significant correlations with subjective ratings of thirst and mood state. PMID- 11876781 TI - The anabolic androgenic steroid, nandrolone decanoate, increases the density of Fos-like immunoreactive neurons in limbic regions of guinea-pig brain. AB - The increased abuse of anabolic androgenic steroids is a major concern because of physiological and psychological side-effects. In some individuals they induce dramatic behavioural changes such as increased aggression, anxiety and depression. The mechanisms behind these behavioural changes are still poorly understood. In order to obtain information on the brain regions affected by anabolic androgenic steroids, the distribution of neurons containing c-Fos, the protein product of the immediate early gene c-fos, and Fos-related antigens was studied following chronic treatment of guinea-pigs with a high dose of nandrolone decanoate (15 mg/kg i.m. daily for 14 days). The behaviour of the guinea-pigs was monitored for 1 h each day. Animals treated with nandrolone exhibited a significantly greater incidence of biting behaviour during the 14 day treatment period than vehicle-treated animals. A significantly greater density of c-Fos and Fos-related antigen-positive neurons was found in the central nucleus of the amygdala, and of Fos-related antigen-positive neurons in the frontal cortex, the shell of the nucleus accumbens and the supraoptic nucleus in nandrolone-treated animals than in vehicle controls. Therefore, nandrolone induced Fos in brain regions involved in stress, behavioural responses and reward. The increased Fos expression in these limbic brain regions is of particular interest in relation to the behavioural changes reported in humans who abuse anabolic androgenic steroids and the abuse potential of these drugs. PMID- 11876782 TI - Physiological properties of central amygdala neurons: species differences. AB - The connections of limbic structures are remarkably constant across species. However, different species express fear in distinct ways. Because the central amygdaloid nucleus is considered an important mediator of fear responses, we compared the physiological properties of central amygdala neurons in guinea pigs, rats and cats using whole-cell recordings in coronal slices in vitro. In guinea pigs, most central medial and lateral neurons displayed an outward rectification that delayed firing onset in response to depolarizing current pulses (hence the designation late-firing neurons). Late-firing neurons were also prevalent in the lateral sector of the cat central nucleus. In contrast, late-firing neurons were rare in the rat central nucleus and the cat central medial nucleus. There, most neurons generated Ni2+-sensitive low-threshold bursts of Na+ spikes and/or displayed pronounced inward rectification in the hyperpolarizing direction. Species differences in passive properties were also observed, but they were mainly related to the relative prevalence of late-firing cells. Indeed, late firing cells had a significantly lower input resistance and more negative membrane potential than other types of central neurons. Thus, there are marked species differences in the physiological properties of central neurons. Because the synaptic responsiveness of neurons is constrained by their physiological properties, our findings raise the possibility that the contrasting behavioural responsiveness observed across species is partly dependent on the species specific physiological properties of central neurons. PMID- 11876783 TI - Glucocorticoids interact with the basolateral amygdala beta-adrenoceptor- cAMP/cAMP/PKA system in influencing memory consolidation. AB - Infusion of a beta-adrenoceptor antagonist into the basolateral nucleus of the amygdala (BLA) blocks memory enhancement induced by systemic or intra-BLA administration of a glucocorticoid receptor (GR) agonist. As there is evidence that glucocorticoids interact with the noradrenergic signalling pathway in activating adenosine 3prime prime or minute,5prime prime or minute-cyclic monophosphate (cAMP), the present experiments examined whether glucocorticoids influence the beta-adrenoceptor--cAMP system in the BLA in modulating memory consolidation. Male, Sprague--Dawley rats received bilateral infusions of atenolol (a beta-adrenoceptor antagonist), prazosin (an alpha1-adrenoceptor antagonist) or Rp-cAMPS (a protein kinase A inhibitor) into the BLA 10 min before inhibitory avoidance training and immediate post-training intra-BLA infusions of the GR agonist, RU 28362. Atenolol and Rp-cAMPS, but not prazosin, blocked 48-h retention enhancement induced by RU 28362. A second series of experiments investigated whether a GR antagonist alters the effect of noradrenergic activation in the BLA on memory consolidation. Bilateral intra-BLA infusions of the GR antagonist, RU 38486, administered 10 min before inhibitory avoidance training completely blocked retention enhancement induced by alpha1-adrenoceptor activation and attenuated the dose--response effects of post-training intra-BLA infusions of clenbuterol (a beta-adrenoceptor agonist). However, the GR antagonist did not alter retention enhancement induced by post-training intra-BLA infusions of 8-Br-cAMP (a synthetic cAMP analogue). These findings suggest that glucocorticoids influence the efficacy of noradrenergic stimulation in the BLA on memory consolidation via an interaction with the beta-adrenoceptor--cAMP cascade, at a locus between the membrane-bound beta-adrenoceptor and the intracellular cAMP formation site. PMID- 11876784 TI - Neural background of glucocorticoid dysfunction-induced abnormal aggression in rats: involvement of fear- and stress-related structures. AB - Glucocorticoid hypofunction is associated with persistent aggression in some psychologically disordered human subjects and, as reported recently, induces abnormal forms of aggression in rats. Here we report on the effects of glucocorticoid hypofunction on aggression-induced neural activation. Rats were adrenalectomized, and implanted with low-release glucocorticoid pellets. After one week recovery, they were challenged by an unfamiliar intruder in their home cage. Neural activation was studied by c-Fos protein immunocytochemistry. Aggressive encounters in controls induced c-Fos activation in all brain areas relevant for the control of aggression (cortex, amygdala, septum, hypothalamus, periaqueductal grey and the locus coeruleus). Very intense c-Fos activation was observed in the medial amygdala, the hypothalamic attack area and the periaqueductal grey matter which constitute a downward stimulatory stream that activates attack behaviour. The experimentally induced glucocorticoid hypofunction dramatically increased attacks targeted towards vulnerable parts of the opponent's body (mainly the head). This abnormal behaviour was not associated with changes in the activation of brain centres involved in the control of aggression. However, the activation of brain centres involved in both the stress response (the parvocellular part of the hypothalamic paraventricular nucleus) and fear reactions (central amygdala) were markedly increased. An acute glucocorticoid treatment abolished both behavioural and neural consequences of glucocorticoid hypofunction. Our data suggest that glucocorticoid hypofunction induced abnormal forms of aggressiveness are related to increased sensitivity to stressors and fear-eliciting stimuli. This assumption is supported by the finding that fearful situations induce attack patterns in intact rats that are similar to those induced by glucocorticoid hypofunction. PMID- 11876785 TI - Vasoactive intestinal polypeptide induces per1 and per2 gene expression in the rat suprachiasmatic nucleus late at night. AB - Circadian rhythms in behaviour and physiology generated by the suprachiasmatic nucleus (SCN) are entrained to the environmental light/dark cycle via the retinohypothalamic tract. How light is able to adjust the endogenous rhythm is not fully understood, but induction of the two clock genes per1 and per2 in the SCN is believed to be important for the adjustment. Recently, it was shown that vasoactive intestinal polypeptide (VIP), a neurotransmitter found in light responsive cells of the SCN, is able to phase shift the circadian rhythm similar to light. In the present study we show by means of an in vitro brain slice model and quantitative in situ hybridization histochemistry that VIP induces both per1 and per2 gene expression in the SCN during late subjective night (CT19). The signalling pathways responsible for the VIP signalling to the clock were investigated using inhibitors of protein kinase A and phospholipase C mediated signalling. Our results demonstrate that both pathways are involved in VIP induced per gene expression and suggest that VIP is important for light-induced phase shift late at night. PMID- 11876786 TI - Haematopoietic progenitor cells from adult bone marrow differentiate into cells that express oligodendroglial antigens in the neonatal mouse brain. AB - Stem cells are self-renewable, pluripotent cells that, in adult life, proliferate by a characteristic asymmetric division in which one daughter cell is committed to differentiation whereas the other remains a stem cell. These cells are also characterized by their ability to differentiate into various cell types under heterotopic environmental influences. In the present study, we have explored the potential of adult haematopoietic bone marrow cells to differentiate into cells of oligodendroglial lineage under physiological, active myelinating conditions. We present evidence of generation of cells expressing oligodendroglial specific markers from a bone marrow subpopulation enriched on adult haematopoietic progenitor cells (CD117+) in vivo after intracerebral transplantation into the neonatal mouse brain. Our results suggest that adult bone marrow cells have the capacity to undergo differentiation from haematopoietic to oligodendroglial cells and add support the validity of bone marrow transplants as an alternative treatment for demyelinating diseases of the CNS including Multiple Sclerosis. PMID- 11876789 TI - Management of chronic hepatitis C--time for a change? PMID- 11876787 TI - NMDA receptors and PSD-95 are found in attachment plaques in cerebellar granular layer glomeruli. AB - N-methyl-D-aspartate (NMDA) receptors mediate long-term changes in excitatory synapses in response to glutamate release. In the cerebellar granular layer, most glutamatergic synapses are formed between mossy terminals and granule cell dendrites, which together with some other components, make up complex glomerular structures. Glomeruli contain numerous attachment plaques (or puncta adherentia), which are sites of adhesion between cells. These structures are found mainly between granule cell dendrites, and probably help maintain the integrity of glomeruli. Attachment plaques contain adhesive proteins such as cadherins. In this study, we show that NMDA receptors are common at these attachment plaques, in addition to being found at synapses. We used four different antibodies to the NMDA receptor subunit, NR1, and another to NR2A/B. In contrast, labelling for an alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA) glutamate receptor antibody was seen only in a few attachment plaques, although AMPA receptors were seen frequently at glomerular synapses. We also show that substantial levels of the NMDA receptor-associated protein, PSD-95, are found in both synapses and attachment plaques. One way that NMDA receptors mediate changes in synapses is through effects on synaptic cadherins, which change their adhesive properties in response to NMDA receptor activation and consequently may alter synaptic function. The presence of NMDA receptors in attachment plaques suggests that these receptors mediate changes in the adhesive properties of these plaques, similar to this function in synapses. PMID- 11876790 TI - Hepatitis C: an epidemiological review. AB - The aim of the study was to analyse the current literature regarding the mode of transmission of HCV and its global prevalence in different groups of people. A systematic review of the literature on the epidemiology of hepatitis C from 1991 to 2000 using computerized bibliographic databases which include Medline, Current Content and Embase. The prevalence of hepatitis C virus (HCV) varies tremendously in different parts of the world, with the highest incidence in the Eastern parts of the globe compared with the Western parts. Furthermore, certain groups of individuals such as intravenous drug users are at increased risk of acquiring this disease irrespective of the geographical location. Although the main route of transmission is via contaminated blood, curiously enough in up to 50% of the cases no recognizable transmission factor/route could be identified. Therefore, a number of other routes of transmission such as sexual or household exposure to infected contacts have been investigated with conflicting results. Hepatitis C infection is an important public health issue globally. Better understanding of routes of transmission will help to combat the spread of disease. In order to prevent a world wide epidemic of this disease, urgent measures are required to (i) develop a strategy to inform and educate the public regarding this disease and (ii) expedite the efforts to develop a vaccine. PMID- 11876791 TI - Quantification and duration of viraemia during hepatitis A infection as determined by real-time RT-PCR. AB - Human transmission studies and molecular techniques have provided evidence that transient viraemia occurs during infection with hepatitis A virus (HAV). However, the duration of its presence and levels during the phases of clinical disease and convalescence has not yet been well studied in human patients. Real-time RT-PCR techniques are increasingly used to quantify RNA viruses for diagnosis and/or research purposes. We have optimized a one-step RT-PCR that contains a dual labelled fluorogenic probe to quantify the 5' noncoding region (5' NCR) of HAV. This method has a dynamic range (5-5 x 10(6) copies). The coefficient of regression of the standard curve was, on average 0.978. Intra-assay CVs% varied from 6.1% to 0.98%, and interassay CVs% from 6.46% to 2.1%. In the currently reported study 41 HAV IgM positive serum samples and 200 serum samples from healthy blood donors were tested by the quantitative RT-PCR method. The mean values on the first day of diagnosis found was 6.38 x 10(5) copies/mL. In a longitudinal study, viraemia persisted for an average of 60 days after clinical onset. These results show that viraemia in HAV infection lasts for many weeks. PMID- 11876792 TI - Apoptosis mediated by the Fas system in the fulminant hepatitis by hepatitis B virus. AB - The pathogenesis of the fulminant hepatitis B is poorly understood and both viral factors and the hosts immune response play a role. Previous studies in liver tissues of patients with chronic hepatitis B showed overexpression of Fas antigen and this was correlated with the activity of the hepatitis. The present study was done to determine the role of Fas in fulminant hepatitis B and the virological characteristics of hepatitis B infection. We studied three patients with fulminant hepatitis B. HBV-DNA was detected by dot-blot hybridization and polymerase chain reaction. The S and C gene were sequenced. Levels of serum soluble Fas antigen were detected by enzymoimmunoassays procedure. Apoptosis was determined by the TUNEL technique. Fas antigen expression was evaluated by a immunoperoxidase method. Ten healthy subjects acted as controls. The three patients showed a high expression of Fas antigen particularly among infiltrating lymphocytes; in these areas we also found many cells with in situ DNA nick labelling signals in the nuclei of most viable hepatocytes. Serum levels of soluble Fas antigen were higher in patients with fulminant hepatitis B than in controls. No specific genome mutations of hepatitis B virus were found. These data suggest that the Fas system involved in the liver injury of patients with fulminant hepatitis B. PMID- 11876793 TI - Quantification of fibrosis progression in patients with chronic hepatitis C using a Markov model. AB - The knowledge of fibrosis progression in chronic hepatitis C and the impact of new treatments on progression is limited by the number of available liver biopsies per patient. Moreover, liver biopsies identify a patient's stage of fibrosis at a given point in time, but cannot quantify the time spent in that stage nor the date of transition to that stage. This paper assesses the potential of Markov modelling to overcome these difficulties. The data from interferon treated (n=185) and untreated patients (n=102) are analysed to illustrate the power of this technique. The model accurately reproduced the distributions of patients in the different fibrosis stages at two subsequent biopsies. A quantification of the role of cofactors in the progression of the disease, and the impact of interferon treatment are given. In subjects who are 40 years old and have been infected for 10 years, the model predicted that 274 of 1000 untreated patients, but only 42 of 1000 treated patients, would progress from F0 or F1 to F3 or F4 fibrosis over the next 5 years. The model also confirms that as age and duration of infection increase, the risk of fibrosis progression increases, while the impact of treatment with interferon decreases. Hence Markov modelling is an accurate tool in the analysis of fibrosis progression in chronic hepatitis C. It will be valuable for the quantification of the effect of new treatments on fibrosis progression in hepatitis C. PMID- 11876794 TI - In situ polymerase chain reaction detection of transfusion-transmitted virus in liver biopsy. AB - The potential role of transfusion-transmitted virus (TTV) infection in determining liver damage is poorly understood and no information exists about TTV replication within hepatocytes. In this study, we assess TTV in situ PCR in liver tissue. Twenty-one patients with different degrees of liver damage were studied by both serum TTV-DNA detection and in situ TTV PCR analysis and extractive PCR in liver biopsy paraffin sections (FFPE). Extractive PCR and in situ PCR detected TTV-DNA both in serum and liver tissue of five patients. The presence of TTV in serum matched with that found in the liver and TTV sequences were never found independently in liver or serum. Four out of five TTV-DNA-positive patients have not other known cause of liver damage while in one a coinfection from HCV was observed. Our data indicate that in situ PCR appears to be a reliable tool for the detection of TTV-DNA in FFPE, and may help detecting unknown origin of liver damage. PMID- 11876795 TI - Biochemical markers of liver fibrosis in patients infected by hepatitis C virus: longitudinal validation in a randomized trial. AB - A liver fibrosis index was recently prospectively validated in a cross-sectional study where patients infected by hepatitis C virus (HCV) had only one biopsy and no longitudinal follow-up. The aim of this study was to retrospectively assess the diagnostic value of this index in patients included in a randomized trial of interferon (IFN) using repeated measurements, two biopsies and hyaluronic acid as a comparative reference. One-hundred and sixty-five patients who had had two interpretable liver biopsies and at least one stored serum sample before IFN treatment were selected. Seventy-eight patients received 3 MU of IFN-alpha thrice weekly for 24 weeks and 87 followed a reinforced regimen for 48 weeks. A fibrosis index combining five biochemical markers (alpha2-macroglobulin, haptoglobin, apolipoprotein A1, gamma-glutamyl transpeptidase (GGT) and total bilirubin adjusted for gender and age) as well as hyaluronic acid was assessed on 461 samples available at baseline, at the end of treatment and at the end of follow up (72 weeks). There was a significant decrease of the fibrosis index score among the 17 sustained virologic responders, from 0.33 +/- 0.06 (mean +/- SE) at baseline to 0.18 +/- 0.06 at 72 weeks in comparison with 92 nonresponders (from 0.41 +/- 0.03 at baseline to 0.44 +/- 0.03 at 72 weeks; P < 0.001) and in comparison with 56 relapsers (from 0.36 +/- 0.03 at baseline to 0.32 +/- 0.03 at 72 weeks; P=0.05). No significant differences were observed for hyaluronic acid.Hence, this fibrosis index could be used as a surrogate marker of the antifibrotic effect of treatments in patients with chronic hepatitis C. PMID- 11876796 TI - Kinetics of serum cytokines reflect changes in the severity of chronic hepatitis C presenting minimal fibrosis. AB - Our aims were to measure the kinetics of serum tumour necrosis alpha (TNF-alpha) and transforming growth factor beta (TGF-beta) levels as markers of progression of disease in nontreated chronic hepatitis C virus (HCV)-infected patients with minimal or no fibrosis and minimal histology activity index (HAI) scores. Our study group consisted of 56 patients diagnosed with minimal (1) or no fibrosis (0) and minimal HAI (0-1) on their first biopsy as defined by Knodell and METAVIR scores. We compared their initial (entry of study) cytokine levels with a group of 103 HCV controls with minimal (0-1) to mild fibrosis (0-3) and mild HAI (5.5). Serum TNF-alpha and TGF-beta levels were measured by enzyme-linked-immunosorbent assay. A significant difference was seen in TNF-alpha levels at baseline in the study group vs. controls. Regardless of their HAI, there was a correlation between TGF-beta and degree of fibrosis. As shown by their biopsies, during the 3 years (from entry to follow up), many of the patients that initially had minimal fibrosis progressed to higher degree of fibrosis. This progression is paralleled by an increase in TGF-beta levels when comparing initial and follow-up levels. In conclusion, serum TNF-alpha reflects the progression of inflammation as seen in liver biopsies and TGF-beta reflects the degree of fibrosis in HCV patients. PMID- 11876797 TI - Hepatitis C virus infection in renal failure patients in the absence of anti hepatitis C virus antibodies. AB - The magnitude and clinical significance of Hepatitis C virus (HCV) infection in dialysis patients is controversial and underestimated. This study was conducted in order to evaluate the correlation between HCV replication and antibody response to HCV in dialysis patients. HCV infection in dialysis patients was evaluated over a period of 3 years and compared to HCV infection in Liver Clinic patients. Sera were collected from 310 dialysis patients and tested for anti-HCV and HCV-RNA. In addition, HCV genotype and HCV viral load were determined in HCV RNA-positive sera. Anti-HCV was detected in 43 (14%) of the dialysis patients. Of these, 37 (86%) were HCV-RNA-positive. Among the 267 HCV-seronegative dialysis patients, 25 (9%) were found to be HCV-RNA-positive in more than one sample during the study. These patients were characterized by low viral load; at least two orders of magnitude lower than in the group of HCV-seropositives. In contrast, in the Liver Clinic patients, HCV-RNA was found exclusively in HCV seropositive patients. Comparison of the genotype pattern in the two groups did not reveal a difference. Our results suggest that HCV infection in dialysis units may be underestimated due to cases of low viral load, depending on the method of RNA extraction and sensitivity of the test used. Low viral load might contribute to the lack of humoral immune response seen in some dialysis patients. PMID- 11876798 TI - Concurrence of hepatitis B surface antibodies and surface antigen: implications for postvaccination control of health care workers. AB - Among 1081 persons testing positive for hepatitis B surface antigen, 106 (10%) tested positive for antibodies to surface antigen (anti-HBs) in the same blood sample. Thirty of these persons were studied in detail: seven tested positive for hepatitis B e-antigen, nine were apparently healthy blood donors, and in 14 chronic infection could be demonstrated in follow-up samples. Frozen samples of 14 persons were available for additional quantitative anti-HBs testing using another anti-HBs assay: three showed no anti-HBs reactivity, seven showed borderline anti-HBs levels (1-5 IU/L), and anti-HBs titres ranged from 23 to 66 IU/L in four HBsAg-positive persons, including an apparently healthy blood donor. Thus, after hepatitis B vaccination of medical personnel, presence of anti-HBs may erroneously suggest immunity, while in fact chronic infection with hepatitis B virus is present. PMID- 11876799 TI - Full-length sequence and functional analysis of hepatitis B virus genome in a virus carrier: a case report suggesting the impact of pre-S and core promoter mutations on the progression of the disease. AB - In chronic hepatitis B virus (HBV) infection, the quiescent immunotolerant phase evolves into the immunoactive phase. The aim of the present study was to clarify the virological alterations relevant to progression. Serial serum samples obtained from a patient with HBV during long-term follow-up were analysed by sequencing of the full-length HBV-DNA using polymerase chain reaction (PCR). In addition, PCR products of HBV genome from each serum sample were transfected into HuH-7 human hepatoma cells for the functional analysis of the transfected viral genomes. Based on the HBV-DNA sequence analysis, the patient had the genotype C virus, and the mutant HBV with common core promoter mutations (T(1762)A(1764)) and deletion of the pre-S region responsible for large surface protein transcription emerged before the onset of hepatitis. When the vigorous host immune response developed (indicated by the flare-up of hepatitis), the mutant HBV containing common core promoter mutations and another pre-S deletion causing lack of the surface protein promoter became predominant. The HBV-DNA sequences, other than pre-S and core promoter regions were identical to the wild-type sequence throughout the study. Transfection of PCR products containing the mutant HBV sequences resulted in increased amounts of intracellular replicative intermediates but the decreased secretion of HBsAg and HBeAg into culture media, suggesting accumulation of nonenveloped viral core particles within the cells. These results indicate that pre-S deletion and core promoter mutations may participate cooperatively in progression of the disease. PMID- 11876800 TI - Three-dimensional ultrasound in gynecology: a critical evaluation. PMID- 11876801 TI - Timing the delivery of the preterm severely growth-restricted fetus: venous Doppler, cardiotocography or the biophysical profile? PMID- 11876803 TI - Fetal ear length measurement: a useful predictor of aneuploidy? AB - OBJECTIVE: To determine the usefulness of short ear length (EL) measurement in the prenatal detection of fetuses with chromosomal abnormalities. DESIGN: Fetal EL measurements, routine biometry and complete anatomic survey for fetal abnormalities were prospectively performed by antenatal sonography. SUBJECTS: One thousand eight hundred and forty-eight patients with singleton pregnancies undergoing genetic amniocentesis in the second or third trimester. METHODS: Complete data for EL, biometry and anatomic survey for major structural abnormalities and minor sonographic markers of chromosomal abnormality were available in 1311 fetuses. Of these, 48 (3.7%) had an abnormal karyotype and 1263 (96.3%) had a normal karyotype. Using an EL measurement of < or = 10th percentile for corresponding gestational age in normal fetuses as abnormal cut-off values, detection rates for chromosomal abnormalities by short EL were determined. RESULTS: Among the 48 abnormal karyotypes, 34 were considered significant, and 11 of these 34 (32.4%) fetuses had short EL. In 14 cases, the karyotypic abnormality was considered non-significant and fetal EL was normal in all cases. Of the 34 fetuses with significant chromosomal abnormalities, six (17.6%) on antenatal sonography had no detectable abnormal findings, other than short EL. An increased biparietal diameter (BPD)/EL ratio of > or = 4.0 was also noted in fetuses with an abnormal karyotype, but the sensitivity and predictive value of increased BPD/EL ratio alone or increased BPD/EL ratio in combination with short EL was no better than the sensitivity and predictive value of short EL alone. A combination of short EL and abnormal ultrasound, however, gave a much higher positive predictive value (46%) for significant chromosomal abnormalities. CONCLUSIONS: Our findings suggest that in women at high risk for fetal chromosomal abnormality, a short fetal EL measurement on prenatal ultrasound, either alone or in combination with other sonographically detectable structural abnormalities, may be a useful parameter in predicting fetal aneuploidy. PMID- 11876804 TI - The development of the fetal vermis: an in-utero sonographic evaluation. AB - OBJECTIVE: To establish a nomogram for fetal vermis measurements during gestation. METHODS: A prospective cross-sectional study of normal singleton pregnancies. Measurements of the fetal vermis width (in the axial plane) and height (in the sagittal plane) were performed by high-resolution transabdominal ultrasonography between 18 and 38 weeks of gestation in 256 fetuses. RESULTS: Adequate vermis measurements were obtained in 256 fetuses. Vermian width and height as a function of gestational age and biparietal diameter were expressed by regression equations and the correlation coefficients were found to be highly statistically significant (P < 0.0001). The normal mean (+/- SD) for each gestational week was defined. CONCLUSIONS: The present data offer the normal range of the vermian measurements throughout gestation. These values may allow intrauterine assessment of the development of the cerebellar vermis, as well as the posterior fossa. PMID- 11876802 TI - The fetal mandible: a 2D and 3D sonographic approach to the diagnosis of retrognathia and micrognathia. AB - OBJECTIVE: To define parameters that enable the objective diagnosis of anomalies of the position and/or size of the fetal mandible in utero. DESIGN: Fetuses at 18 28 gestational weeks were examined by two- and three-dimensional ultrasound. The study included normal fetuses and fetuses with syndromes associated with known mandible pathology: Pierre Robin sequence or complex (n = 8); hemifacial microsomia (Treacher-Collins syndrome, n = 3); postaxial acrofacial dysostosis (n = 1). Fetuses with Down syndrome (n = 8) and cleft lip and palate without Pierre Robin sequence or complex (n = 18) were also studied. Retrognathia was assessed through the measurement of the inferior facial angle, defined on a mid-sagittal view, by the crossing of: 1) the line orthogonal to the vertical part of the forehead at the level of the synostosis of the nasal bones (reference line); 2) the line joining the tip of the mentum and the anterior border of the more protruding lip (profile line). Micrognathia was assessed through the calculation of the mandible width/maxilla width ratio on axial views obtained at the alveolar level. Mandible and maxilla widths were measured 10 mm posteriorly to the anterior osteous border. RESULTS: In normal fetuses, the inferior facial angle was constant over the time span studied. The mean (standard deviation) value of the inferior facial angle was 65.5 (8.13) degree. Consequently, an inferior facial angle value below 49.2 degree (mean - 2 standard deviations) defined retrognathism. All the fetuses with syndromes associated with mandible pathology had inferior facial angle values below the cut-off value. Using 49.2 degree or the rounded-up value of 50 degree as a cut-off point, the inferior facial angle had a sensitivity of 1.0, a specificity of 0.989, a positive predictive value of 0.750 and a negative predictive value of 1.0 to predict retrognathia. In normal fetuses, the mandible width/maxilla width ratio was constant over the time interval studied. The mean (standard deviation) value was 1.017 (0.116). Consequently, a mandible width/maxilla width ratio < 0.785 defined micrognathism. Mandible width/maxilla width ratio values were below this cut-off point in eight and in the normal range in four fetuses with syndromes associated with mandible pathology. CONCLUSIONS: Retrognathia and micrognathia are conditions that can be separately assessed. The use of inferior facial angle and mandible width/maxilla width ratio should help sonographic recognition and characterization of fetal retrognathic and micrognathic mandibles in utero. PMID- 11876805 TI - Temporal sequence of abnormal Doppler changes in the peripheral and central circulatory systems of the severely growth-restricted fetus. AB - OBJECTIVE: To identify the temporal sequence of abnormal Doppler changes in the fetal circulation in a subset of early and severely growth-restricted fetuses. METHODS: This was a prospective observational study in a tertiary care/teaching hospital. Twenty-six women who were diagnosed with growth-restricted fetuses by local standards before 32 weeks' gestation and who had abnormal uterine and umbilical artery Doppler velocimetry were enrolled onto the study. To compare Doppler changes as a function of time, pulsed-wave Doppler ultrasound was performed on five vessels in the fetal peripheral and central circulations. Doppler examinations were performed twice-weekly and on the day of delivery if the fetal heart rate tracing became abnormal. Doppler indices were scored as abnormal when their values were outside the local reference limits on two or more consecutive measurements. Biometry for assessment of fetal growth was performed every 2 weeks. Computerized fetal heart rates were obtained daily. Delivery was based on a non-reactive fetal heart rate tracing and not on Doppler information. Patients with a severely growth-restricted fetus who were delivered for maternal indications such as pre-eclampsia were excluded. Perinatal outcome endpoints included: intrauterine death, gestational age at delivery, newborn weight, central nervous system damage of grade 2 or greater, intraventricular hemorrhage and neonatal mortality. RESULTS: Mean gestational age and newborn weight at delivery were 29 (standard deviation (SD), 2) weeks and 818 (SD, 150) g, respectively. The sequence of Doppler velocimetric changes was described by onset time cumulative curves that showed two time-related events. First, for each vessel there was a progressive increase in the percent of fetuses developing a Doppler abnormality. Second, severely growth-restricted fetuses followed a progressive sequence of acquiring Doppler abnormalities which were categorized into 'early' and 'late' Doppler changes. Early changes occurred in peripheral vessels (umbilical and middle cerebral arteries; 50% of patients affected 15-16 days prior to delivery). Late changes included umbilical artery reverse flow, and abnormal changes in the ductus venosus, aortic and pulmonary outflow tracts (50% of patients affected 4-5 days prior to delivery). The time interval between the occurrence of early and late changes was significantly different (P < 0.0001) and late changes were significantly associated with perinatal death (P < 0.01). CONCLUSIONS: Doppler velocimetry abnormalities develop in different vessels of the severely growth-restricted fetus in a sequential fashion. Late changes in vascular adaptation by the severely growth-restricted fetus are the best predictor of perinatal death. PMID- 11876806 TI - Retrograde net blood flow in the aortic isthmus in relation to human fetal arterial and venous circulations. AB - OBJECTIVES: To characterize changes in the human fetal arterial and venous circulations associated with retrograde aortic isthmus net blood flow. METHODS: Study groups consisted of fetuses with placental insufficiency and/or fetal growth restriction and either antegrade (Group 1; n = 18) or retrograde (Group 2; n = 11) net blood flow in the aortic isthmus. The control group comprised 31 fetuses in uncomplicated pregnancies. Pulsatility indices of the umbilical, middle cerebral and proximal pulmonary arteries and the descending aorta, and pulsatility indices for veins of the ductus venosus and inferior vena cava were calculated. Right and left ventricular fractional shortenings were ascertained. The coronary artery blood flow was visualized and the presence of tricuspid regurgitation was noted. RESULTS: In the study groups, the umbilical artery and descending aorta pulsatility indices were significantly higher (P < 0.05), and those of the middle cerebral artery lower (P < 0.001), than in the control group, with no difference between the two study groups. The proximal pulmonary artery pulsatility index was significantly higher in Group 2 (P < 0.001) than in Group 1 and the control group. In Group 2, the right ventricular fractional shortening was significantly lower (P < 0.01) than in Group 1. Coronary artery blood flow was visualized significantly more often (P < 0.03) and tricuspid regurgitation was present more frequently (P < 0.003) in Group 2 than in Group 1. In Group 2, the ductus venosus pulsatility index for veins was significantly higher than in Group 1 (P < 0.01) and the control group (P < 0.01), with no difference in the inferior vena cava pulsatility index for veins. CONCLUSIONS: Fetuses with retrograde aortic isthmus net blood flow demonstrate a rise in right ventricular afterload and increased pulsatility in ductus venosus blood velocity waveforms. PMID- 11876807 TI - Fetal aortic isthmus growth and morphology in late gestation. AB - OBJECTIVE: To establish normal values for fetal aortic isthmus diameter in late gestation and to identify any changes in aortic isthmus dimensions and morphology in pathological conditions. METHODS: In this prospective study, the fetal aortic isthmus was evaluated in 110 low-risk pregnant women at between 30 and 40 weeks of gestation and 42 pregnant women who were at high risk for congenital heart defects. From coronal echocardiographic images of the connection between the aorta and ductus arteriosus, the internal diameter of the aorta was measured at the middle of the isthmus, at the point of the isthmus just proximal to the entry of the ductus arteriosus and at the descending aorta below the entry of the ductus arteriosus. RESULTS: Correlation coefficients for the diameter of each aortic segment when related to gestational age varied from r = 0.60 to r = 0.80 (P < 0.001 for each), and growth curves were derived from the third and 97th percentiles about each linear regression analysis. The mean and the third percentile for the ratio of the isthmus just proximal to the entry of the ductus arteriosus to the middle of the isthmus were 1 and 0.81. In one fetus of the high risk patients, a contraductal shelf and the accompanying area of tubular isthmic hypoplasia were suspected and a diagnosis of coarctation of the aorta was subsequently confirmed after birth. In two fetuses with growth restriction and one fetus with intestinal atresia, the isthmus diameters were below the third percentile but the ratios of the isthmus end to the middle of the aortic isthmus were all normal and no cardiac anomalies were detected after birth. CONCLUSIONS: We could establish normal values for aortic isthmus diameters in late gestation from a coronal view and identify even minimal changes in aortic isthmus dimensions and morphology in pathological conditions. PMID- 11876808 TI - Flecainide in the intrauterine treatment of fetal supraventricular tachycardia. AB - OBJECTIVES: To assess the efficacy of flecainide in the intrauterine treatment of fetal supraventricular tachycardia (SVT) with 1 : 1 atrioventricular conduction. DESIGN: Twenty fetuses (21-35 weeks of gestation) with SVT ranging between 215 and 280 bpm were analyzed retrospectively. Fetuses received flecainide and digoxin as either first, second or third line therapy. Intracardiac blood flow, venous Doppler waveforms and cardiotocograms were evaluated before and after drug induced conversion to sinus rhythm. RESULTS: After initiation of combined flecainide and digoxin therapy, the median time interval until final conversion to sinus rhythm was 5 days (range, 0-14 days). The majority of fetuses (n = 15; 75%) converted to sinus rhythm within 7 days of treatment, whereas the remaining five (25%) showed initial reduction of the heart rate to 160-215 bpm over several days, with restoration of a triphasic venous blood flow pattern before late conversion within 7-14 days after initiation of flecainide treatment. One of these fetuses showed a decrease in fetal heart rate to 160-190 bpm without conversion to sinus rhythm but with resolution of hydrops. All fetuses survived. CONCLUSIONS: Flecainide is safe and highly effective in the intrauterine treatment of hydropic fetuses with supraventricular tachycardia. Conversion into sinus rhythm can be expected 72 h after initiation of therapy but may take up to 14 days. Therefore therapy should be continued beyond 72 h, especially when an initial decrease of fetal heart rate is observed which may represent an early therapeutic response. PMID- 11876810 TI - Routine ultrasound screening in diabetic pregnancies. AB - OBJECTIVES: To assess the detection rate of congenital fetal malformations and specific problems related to routine ultrasound screening in women with pre existing diabetes. METHODS: A retrospective study was carried out to assess the performance of routine ultrasound screening in women with pre-existing diabetes (Types 1 and 2) within a tertiary institution. The incidence, type and risk factors for congenital fetal malformations were determined. The detection rate of fetal anomalies for diabetic women was compared with that for the low-risk population. Factors affecting these detection rates were evaluated. RESULTS: During the study period, 12 169 low-risk pregnant women and 130 women with pre existing diabetes had routine ultrasound screening performed within the institution. A total of 10 major anomalies (7.7%) and three minor anomalies (2.3%) were present in the fetuses of the diabetic women. Central nervous system and cardiovascular system anomalies accounted for 60% of the major anomalies. Periconceptional hemoglobin A1c of more than 9% was associated with a high prevalence of major anomalies (143/1000). Women who had fetuses with major anomalies had a significantly higher incidence of obesity (78% vs. 37%; P < 0.05). Ultrasound examination of these diabetic pregnancies showed high incidences of suboptimal image quality (37%), incomplete examinations, and repeat examinations (17%). Compared to the 'low-risk' non-diabetic population from the same institution, the relative risk for a major congenital anomaly among the diabetic women was 5.9-fold higher (95% confidence interval, 2.9-11.9). The detection rate for major fetal anomalies was significantly lower for diabetic women (30% vs. 73%; P < 0.01), and the mean body mass index for the diabetic group was significantly higher (29 vs. 23 kg/m2; P < 0.001). CONCLUSION: The incidence of congenital anomalies is higher in diabetic pregnancies. Unfortunately, the detection rate for fetal anomalies by antenatal ultrasound scan was significantly worse than that for the low-risk population. This is likely to be related to the maternal body habitus and unsatisfactory examinations. Methods to overcome these difficulties are discussed. PMID- 11876809 TI - Impaired uteroplacental blood flow in pregnancies complicated by falciparum malaria. AB - OBJECTIVE: In endemic areas, maternal malaria infection is usually asymptomatic. However, it is known that infected maternal erythrocytes sequester in the intervillous space of the placenta. There is a strong association between placental malaria infection and both low birth weight (LBW) and severe maternal anemia. We aimed to determine whether impaired uteroplacental blood flow might account for the low infant birth weight associated with maternal falciparum malaria infection. METHODS: This observational study was carried out during a large double-blind, randomized, controlled trial of an antimalarial drug intervention for primigravidae. Nine hundred and ninety-five women were recruited from the antenatal clinic at a district hospital on the Kenya coast and had at least one Doppler ultrasound scan. Uterine artery resistance index and the presence or absence of a diastolic notch were recorded. In the third trimester, blood was taken for hemoglobin and malaria film. RESULTS: Malaria infection at 32 35 weeks of gestation was associated with abnormal uterine artery flow velocity waveforms on the day of blood testing (relative risk (RR) 2.11, 95% confidence interval (CI) 1.24-3.59, P = 0.006). This association persisted after controlling for pre-eclampsia. Impaired uteroplacental blood flow in the women studied was also predictive of poor perinatal outcome, including low birth weight, preterm delivery and perinatal death. The risk of preterm delivery in women with histological evidence of past placental malaria infection was more than twice that of women without infection (RR 2.33, 95% CI 1.31-4.13, P = 0.004). CONCLUSIONS: Uteroplacental hemodynamics are altered in the presence of maternal falciparum malaria infection. This may account for some of the excess of LBW babies observed in malaria endemic areas. Strategies that prevent or clear placental malaria may confer perinatal benefit through preservation of placental function. PMID- 11876811 TI - Uterine artery resistance and anxiety in the second trimester of pregnancy. AB - OBJECTIVE: To investigate the association between maternal anxiety and uterine artery resistance index (RI) at 20 weeks of gestation. METHODS: Uterine artery blood flow was assessed using color Doppler ultrasound and maternal anxiety was measured using the Hospital Anxiety and Depression (HAD) scale in 96 healthy primigravid women attending consecutively for their routine 20-week anomaly scan. RESULTS: The mean uterine artery RI was 0.54 (95% confidence interval, 0.52-0.56) and the median HAD anxiety score was 6 (range, 0-20). There was no association between RI and anxiety scores (r = 0.09, P = 0.36). Women scoring as definite cases of anxiety did not have a significantly elevated uterine artery RI or increased frequency of waveform notching compared to women with doubtful or no anxiety. CONCLUSIONS: The data of this study do not suggest a significant association between maternal anxiety and uterine artery RI at 20 weeks of gestation in healthy primigravid women with normally developing pregnancies. A prospective cohort study would be useful to determine the nature of the relationship between maternal anxiety, alteration in uterine artery blood flow and abnormal pregnancy outcome. PMID- 11876812 TI - Unilateral multicystic dysplastic kidney: a combined pre- and postnatal assessment. AB - OBJECTIVE: To review the prenatal assessment of associated renal pathology, non renal pathology and renal biometry, fetal outcome and postnatal urological management in the presence of unilateral fetal multicystic dysplastic kidney. METHODS: A total of 38 singleton pregnancies with fetal unilateral multicystic dysplastic kidney was studied over a 13-year period. Prenatally, fetal biometry, including head and abdominal circumferences and largest longitudinal diameter of the affected and contralateral kidneys, was performed. The amount of amniotic fluid was assessed. Fetal karyotyping was offered in cases of contralateral renal or non-renal pathology. A MAG 3 scan and voiding cystogram was performed approximately 4 weeks after delivery to establish renal function and to exclude urinary reflux. RESULTS: Unilateral fetal multicystic dysplastic kidney was left sided in 53% and right-sided in 47% of cases. The fetus was male in 63% and female in 37% of cases. Associated renal and non-renal pathology existed in 21% and 5% of cases, respectively. The fetal karyotype in these subsets was always normal. The longitudinal diameter of the multicystic dysplastic kidney was above the 95th centile in 87%. There was polyhydramnios in three cases and oligohydramnios in two cases. The prematurity rate was 16%. Postnatal examination revealed a non-functional multicystic kidney in 87% (33/38) of cases. Following surgical removal of the affected kidney, these infants progressed normally. Of the remaining five infants, four died because of associated anomalies and one infant developed normally without surgery. CONCLUSIONS: Fetal outcome is determined by associated renal and/or non-renal structural pathology and not by the size/location of the unilateral multicystic dysplastic kidney or amniotic fluid volume. PMID- 11876813 TI - A new mathematical formula for predicting long bone length in early pregnancy. AB - OBJECTIVE: To propose new mathematical formulae to estimate fetal long bone biometry in early pregnancy and to establish their efficacy in comparison to previously constructed mathematical formulae. METHODS: A study population of 1960 singleton euploid fetuses was referred for transvaginal ultrasound examinations between 71 and 112 days of gestation prior to genetic amniocentesis. To determine the relationship between the biparietal diameter and long bone length, a sample group of 400 randomly chosen normal fetuses was evaluated. Regression equations were derived, then tested in the remaining 1560 control fetuses and compared with previously reported mathematical formulae by other authors. Mean absolute error, mean absolute percentage error and mean systematic error with their standard deviations were calculated. RESULTS: The relationships between femur or humerus length vs. biparietal diameter (BPD) and gestational age (GA) were, respectively: expected femur length = -16.92108 + 0.4569402 x BPD + 0.171617 x GA (P < 0.001) and expected humerus length = -16.28531 + 0.4283019 x BPD + 0.1696017 x GA (P < 0.001). The confidence intervals of the predicted values for different values of biparietal diameter and gestational age and confidence intervals for the regression coefficients, such as the distribution of the residuals, are given. All previous formulae obtained by transabdominal ultrasound demonstrated an overestimation of expected long bones measurements; this was reduced using different formulae obtained in early pregnancy. Using our mathematical formulae, the mean absolute percentage error and the mean systematic error in estimating femur and humerus length were very low (11.15% and -2.02%; 10.59% and -1.74%, respectively). CONCLUSIONS: The new ultrasonographic morphometric models derived from transvaginal measurements in early pregnancy show a good reliability in estimating fetal long bone length. PMID- 11876814 TI - The importance of quality management in fetal measurement. AB - OBJECTIVES: The aims of this study were to evaluate factors contributing to inaccuracy in fetal measurements and to assess the clinical importance of measurement quality. METHODS: One hundred images of biparietal diameter (BPD), head circumference (HC) and abdominal circumference (AC) measurements were collected from six centers (1800 measurements); the proportion meeting quality criteria was assessed. Four hundred images of AC were collected from one center, each image measured by ellipse fitting and tracing methods; clinical agreement between the methods was assessed. Fetal weight estimation (EFW) errors were compared between quality controlled and non-quality controlled studies. Images of three ACs on each of 400 fetuses were collected; where one measurement failed to meet quality criteria, it was compared with an optimal measurement on the same fetus. RESULTS: Eighty-nine percent, 87% and 60% of BPD, HC and AC, respectively, met all quality criteria. Limits of agreement between ellipse and traced AC were 4.7 mm to 12.5 mm; 22% of sections were non-elliptical. EFW errors were significantly different but were confounded by differences in time to delivery. Limits of agreement between optimal and suboptimal AC measurements were -15.1 mm to 7.7 mm. CONCLUSIONS: AC quality criteria are less easily recognized and obtained than those for head measurements; training, adherence to protocols and audit are important. Differences between ellipse and traced AC may not justify the use of separate charts; the number of non-elliptical sections suggests that ellipse fitting is not appropriate. Comparison between EFW errors is not a suitable tool for audit. Failure to meet quality criteria results in clinically significant errors. PMID- 11876816 TI - Prenatal diagnosis of sacrococcygeal teratomas by two- and three-dimensional ultrasound. AB - We suspected two fetuses of having sacrococcygeal teratomas following initial two dimensional ultrasound scanning at 18 and 22 weeks' gestation. The fetuses were then scanned with three-dimensional multiplanar surface and three-dimensional orthogonal planar ultrasound to establish a definitive diagnosis. Although we made a presumptive diagnosis in both cases after initial two-dimensional scanning, with three-dimensional ultrasound we were better able to define the degree of involvement of the sacrum and other pelvic structures of prognostic importance. PMID- 11876815 TI - Prenatal detection of mesoblastic nephroma by sonography and magnetic resonance imaging. AB - Congenital mesoblastic nephroma is the most common neonatal kidney tumor and surgical excision is almost always curative. We report the prenatal detection of congenital mesoblastic nephroma by sonography and magnetic resonance imaging. After birth, a right nephrectomy was performed and the baby recovered well. PMID- 11876817 TI - Prenatal diagnosis of isolated unilateral pulmonary agenesis in the second trimester. AB - Unilateral pulmonary agenesis is a very rare developmental malformation that is often associated with other anomalies including non-immune hydrops. We describe a case of isolated unilateral pulmonary agenesis diagnosed in the second trimester by gray-scale and color Doppler ultrasound. PMID- 11876818 TI - Transvaginal sonographic appearance of the cerebellar vermis at 14-16 weeks' gestation. PMID- 11876819 TI - Prenatal diagnosis of congenital mesoblastic nephroma. PMID- 11876820 TI - Thrombosis of umbilical vein varix. PMID- 11876822 TI - Correlating overrepresented upstream motifs to gene expression: a computational approach to regulatory element discovery in eukaryotes. AB - BACKGROUND: Gene regulation in eukaryotes is mainly effected through transcription factors binding to rather short recognition motifs generally located upstream of the coding region. We present a novel computational method to identify regulatory elements in the upstream region of eukaryotic genes. The genes are grouped in sets sharing an overrepresented short motif in their upstream sequence. For each set, the average expression level from a microarray experiment is determined: If this level is significantly higher or lower than the average taken over the whole genome, then the overerpresented motif shared by the genes in the set is likely to play a role in their regulation. RESULTS: The method was tested by applying it to the genome of Saccharomyces cerevisiae, using the publicly available results of a DNA microarray experiment, in which expression levels for virtually all the genes were measured during the diauxic shift from fermentation to respiration. Several known motifs were correctly identified, and a new candidate regulatory sequence was determined. CONCLUSIONS: We have described and successfully tested a simple computational method to identify upstream motifs relevant to gene regulation in eukaryotes by studying the statistical correlation between overepresented upstream motifs and gene expression levels. PMID- 11876824 TI - The cloning, genomic organization and tissue expression profile of the human DLG5 gene. AB - BACKGROUND: Familial atrial fibrillation, an autosomal dominant disease, was previously mapped to chromosome 10q22. One of the genes mapped to the 10q22 region is DLG5, a member of the MAGUKs (Membrane Associated Gyanylate Kinase) family which mediates intracellular signaling. Only a partial cDNA was available for DLG5. To exclude potential disease inducing mutations, it was necessary to obtain a complete cDNA and genomic sequence of the gene. METHODS: The Northern Blot analysis performed using 3' UTR of this gene indicated the transcript size to be about 7.2 KB. Using race technique and library screening the entire cDNA was cloned. This gene was evaluated by sequencing the coding region and splice functions in normal and affected family members with familial atrial fibrillation. Furthermore, haploid cell lines from affected patients were generated and analyzed for deletions that may have been missed by PCR. RESULTS: We identified two distinct alternately spliced transcripts of this gene. The genomic sequence of the DLG5 gene spanned 79 KB with 32 exons and was shown to have ubiquitous human tissue expression including placenta, heart, skeletal muscle, liver and pancreas. CONCLUSIONS: The entire cDNA of DLG5 was identified, sequenced and its genomic organization determined. PMID- 11876823 TI - Molecular diversity of phospholipase D in angiosperms. AB - BACKGROUND: The phospholipase D (PLD) family has been identified in plants by recent molecular studies, fostered by the emerging importance of plant PLDs in stress physiology and signal transduction. However, the presence of multiple isoforms limits the power of conventional biochemical and pharmacological approaches, and calls for a wider application of genetic methodology. RESULTS: Taking advantage of sequence data available in public databases, we attempted to provide a prerequisite for such an approach. We made a complete inventory of the Arabidopsis thaliana PLD family, which was found to comprise 12 distinct genes. The current nomenclature of Arabidopsis PLDs was refined and expanded to include five newly described genes. To assess the degree of plant PLD diversity beyond Arabidopsis we explored data from rice (including the genome draft by Monsanto) as well as cDNA and EST sequences from several other plants. Our analysis revealed two major PLD subfamilies in plants. The first, designated C2-PLD, is characterised by presence of the C2 domain and comprises previously known plant PLDs as well as new isoforms with possibly unusual features catalytically inactive or independent on Ca2+. The second subfamily (denoted PXPH-PLD) is novel in plants but is related to animal and fungal enzymes possessing the PX and PH domains. CONCLUSIONS: The evolutionary dynamics, and inter-specific diversity, of plant PLDs inferred from our phylogenetic analysis, call for more plant species to be employed in PLD research. This will enable us to obtain generally valid conclusions. PMID- 11876825 TI - Ligamentous rupture of the ACL associated with dislocated fracture of the proximal tibial physis in a 12-year-old boy. AB - BACKGROUND: Dislocated fracture of the proximal physeal plate of the tibia with or without metaphyseal fragment is rare in children. This unusual fracture classically excludes rupture of the anterior cruciate ligament due to the ligament's stability. A combination of both injuries has not been previously published in the literature. CASE PRESENTATION: The authors report the case of a 12-year-old boy who presented with a dislocated fracture (Salter-Harris II) of the proximal tibia combined with ligamentous rupture of the anterior cruciate ligament after a sporting accident. PMID- 11876826 TI - Plasma proteins in a standardised skin mini-erosion (I): permeability changes as a function of time. AB - BACKGROUND: A standardised technique using a suction-induced mini-erosion that allows serial sampling of dermal interstitial fluid (IF) for 5 to 6 days has been described. In the present study, we studied permeability changes as a function of time. METHODS: We examined IF concentrations of total protein concentration and the concentration of insulin (6.6 kDa), prealbumin (55 kDa), albumin (66 kDa), transferrin (80 kDa), IgG (150 kDa) and alpha-2-macroglobulin (720 kDa) as a function of time, using an extraction pressure of 200 mmHg below atmospheric. RESULTS: At 0 h after forming the erosion, mean total IF protein content (relative to plasma) was 26 +/- 13% (SD). For the individual proteins, the relative mean concentrations were 65 +/- 36% for insulin, 48 +/- 12% for albumin, 30 +/- 19% for transferrin, 31 +/- 15%for IgG and 19.5 +/- 10% for alpha-2 macroglobulin. At 24 h, the total IF protein content was higher than at 0 h (56 +/- 26% vs 26 +/- 13%; p < 0.05, diff: 115%), as were some of the individual protein concentrations: prealbumin (50 +/- 24 vs 25 +/- 13%; p < 0.05), albumin (68 +/- 21 vs 48 +/- 12%; p < 0.05) and IgG (55 +/- 30 vs 31 +/- 15%; p = 0.05). ln the interval 24 h to 96 h the concentrations were relatively unchanged. CONCLUSIONS: The results indicate that fluid sampled at 0 h after forming the erosion represents dermal IF before the full onset of inflammation. From 24 h onward, the sampled fluid reflects a steady state of increased permeability induced by inflammation. This technique is promising as a tool for clinically sampling substances that are freely distributed in the body and as a model for studying inflammation and vascular permeability. PMID- 11876828 TI - Effects of herbivory on the reproductive effort of 4 prairie perennials. AB - BACKGROUND: Herbivory can affect every aspect of a plant's life. Damaged individuals may show decreased survivorship and reproductive output. Additionally, specific plant species (legumes) and tissues (flowers) are often selectively targeted by herbivores, like deer. These types of herbivory influence a plant's growth and abundance. The objective of this study was to identify the effects of leaf and meristem removal (simulated herbivory within an exclosure) on fruit and flower production in four species (Rhus glabra, Rosa arkansana, Lathyrus venosus, and Phlox pilosa) which are known targets of deer herbivory. RESULTS: Lathyrus never flowered or went to seed, so we were unable to detect any treatment effects. Leaf removal did not affect flower number in the other three species. However, Phlox, Rosa, and Rhus all showed significant negative correlations between seed mass and leaf removal. Meristem removal had a more negative effect than leaf removal on flower number in Phlox and on both flower number and seed mass in Rosa. CONCLUSIONS: Meristem removal caused a greater response than defoliation alone in both Phlox and Rosa, which suggests that meristem loss has a greater effect on reproduction. The combination of leaf and meristem removal as well as recruitment limitation by deer, which selectively browse for these species, is likely to be one factor contributing to their low abundance in prairies. PMID- 11876827 TI - Probing substrate binding to metallo-beta-lactamase L1 from Stenotrophomonas maltophilia by using site-directed mutagenesis. AB - BACKGROUND: The metallo-beta-lactamases are Zn(II)-containing enzymes that hydrolyze the beta-lactam bond in penicillins, cephalosporins, and carbapenems and are involved in bacterial antibiotic resistance. There are at least 20 distinct organisms that produce a metallo-beta-lactamase, and these enzymes have been extensively studied using X-ray crystallographic, computational, kinetic, and inhibition studies; however, much is still unknown about how substrates bind and the catalytic mechanism. In an effort to probe substrate binding to metallo beta-lactamase L1 from Stenotrophomonas maltophilia, nine site-directed mutants of L1 were prepared and characterized using metal analyses, CD spectroscopy, and pre-steady state and steady state kinetics. RESULTS: Site-directed mutations were generated of amino acids previously predicted to be important in substrate binding. Steady-state kinetic studies using the mutant enzymes and 9 different substrates demonstrated varying Km and kcat values for the different enzymes and substrates and that no direct correlation between Km and the effect of the mutation on substrate binding could be drawn. Stopped-flow fluorescence studies using nitrocefin as the substrate showed that only the S224D and Y228A mutants exhibited weaker nitrocefin binding. CONCLUSIONS: The data presented herein indicate that Ser224, Ile164, Phe158, Tyr228, and Asn233 are not essential for tight binding of substrate to metallo-beta-lactamase L1. The results in this work also show that Km values are not reliable for showing substrate binding, and there is no correlation between substrate binding and the amount of reaction intermediate formed during the reaction. This work represents the first experimental testing of one of the computational models of the metallo-beta lactamases. PMID- 11876829 TI - Accuracy of prognosis estimates by four palliative care teams: a prospective cohort study. AB - BACKGROUND: Prognosis estimates are used to access services, but are often inaccurate. This study aimed to determine the accuracy of giving a prognosis range. METHODS AND MEASUREMENTS: A prospective cohort study in four multi professional palliative care teams in England collected data on 275 consecutive cancer referrals who died. Prognosis estimates (minimum - maximum) at referral, patient characteristics, were recorded by staff, and later compared with actual survival. RESULTS: Minimum survival estimates ranged <1 to 364 days, maximum 7 - 686 days. Mean patient survival was 71 days (range 1 - 734). In 42% the estimate was accurate, in 36% it was over optimistic and in 22% over pessimistic. When the minimum estimate was less than 14 days accuracy increased to 70%. Accuracy was related, in multivariate analysis, to palliative care team and (of borderline significance) patient age. CONCLUSIONS: Offering a prognosis range has higher levels of accuracy (about double) than traditional estimates, but is still very often inaccurate, except very close to death. Where possible clinicians should discuss scenarios with patients, rather than giving a prognosis range. PMID- 11876830 TI - Evaluation of a risk factor survey with three assessment methods. AB - This paper describes the evaluation of questions on a cancer risk factor survey using three different methods: dataset response patterns, qualitative feedback, and questionnaire appraisal. These methods addressed the survey data, procedures and questions. The three methods identified similar issues but also made unique contributions. Dataset response patterns showed missing and out-of-range data, an order effect, and mixed coding. Qualitative feedback revealed lack of clarity, sensitive topics, technical or undefined terms, failure to hear all response options, overlapping response options (as perceived by respondents), coding problems and recall difficulties. Questionnaire appraisal showed technical or undefined terms, complex syntax, hidden definitions, and ambiguous wording. The survey assessment methods described here can improve data quality, especially when limited time and resources preclude in-depth questionnaire development. PMID- 11876831 TI - Environmental tobacco smoke and deaths from coronary heart disease in Canada. AB - A series of recent meta-analyses have concluded that non-smokers who live with smokers face an elevated risk of coronary heart disease (CHD). In this study, we estimated the number of CHD deaths among non-smokers attributable to environmental tobacco smoke (ETS) exposure in their homes. Population attributable risk estimates suggest that in 1997 over 800 Canadians died of CHD caused by passive exposure to ETS. This figure is likely an underestimation of the total number of CHD deaths attributable to ETS, since our study did not estimate the number of deaths among non-smokers caused by ETS exposure in the workplace. However, this partial picture can still help to highlight the burden of disease resulting from this pervasive involuntary environmental exposure. PMID- 11876832 TI - Estimating the population at risk for Ontario Workplace Safety and Insurance Board-covered injuries or diseases. AB - Difficulty in quantifying the population at risk for a work-related injury or disease limits the usefulness of workers' compensation data for surveillance. This article presents a method of obtaining estimates of the Ontario Workplace Safety and Insurance Board (OWSIB)- covered workforce using the Canadian Labour Force Survey (LFS). The method involves extracting that class of worker most likely to be insured by the OWSIB and using actual hours worked to estimate full time equivalents at risk. Compared to population at risk estimates readily available from published tables, the refined crude estimate was 26% lower and ranged from 15 to 79% lower depending on the age group. The percentage decrease from published estimates was generally greater for women compared to men, particularly in the 25 to 39 year age categories. Consequently, the method of deriving population at risk estimates should be considered when comparing rates across sexes, ages, industries or occupations. PMID- 11876833 TI - Under-reporting of maternal mortality in Canada: a question of definition. AB - In Canada, maternal mortality reporting is based on information contained on death certificates. To examine the extent to which maternal deaths are under reported in Canada and whether this is likely to change under the 10th revision of the International Classification of Diseases (ICD), we linked live birth and stillbirth registrations to In Canada death registrations of women aged 10 to 50 for 1988 through 1992. We reviewed the death certificates of women found to have died while pregnant or within a year of the termination of pregnancy. The officially reported maternal mortality ratio for the study years was 3.7 deaths per 100,000 live births. Depending on whether we included deaths not certified as maternal deaths at the time the deaths occurred, revised ratios under ICD-9 ranged from 4.9 to 5.1 per 100,000 live births for deaths from direct obstetric causes and from 0.5 to 1.2 per 100,000 live births for deaths from indirect obstetric causes. Reflecting changes in classification criteria, revised ratios under ICD-10 were lower than those under ICD-9 for deaths from direct obstetric causes - ranging from 3.9 to 4.1 per 100,000 live births - and higher for deaths from indirect obstetric causes ranging from 2.0 to 3.0 per 100,000 live births. Of deaths from direct obstetric causes, those from cerebrovascular disease were the most numerous and also the most likely to be underreported. Deaths from pulmonary embolism and indirect obstetric causes were the next most likely to be underreported. In a companion article we report an investigation of whether deaths from causes not directly related to pregnancy -such as injury, infectious disease and epilepsy - are more or less likely to occur among pregnant and recently pregnant women. PMID- 11876834 TI - Cause-specific mortality during and after pregnancy and the definition of maternal death. AB - As part of a study to determine whether maternal mortality in Canada is under- reported, we explored the validity of including deaths not directly related to pregnancy. We linked live birth and stillbirth registrations to death registrations of women of reproductive age from 1988 through 1992. We calculated standardized mortality ratios, by cause, from deaths in women known to have been pregnant and deaths in same-aged women not known to have been pregnant within the same time period. Women known to have been pregnant were approximately half as likely to die as would be expected in each of two six-month time periods: from 20 weeks gestation to 42 days postpartum (SMR 0.4, 95% CI 0.3-0.5), and from 42 days to 225 days postpartum (SMR 0.5, 95% CI 0.5-0.6). Furthermore, pregnant and recently pregnant women were not more likely to die from specific causes, with the exception of diseases of the arteries, arterioles, and capillaries (SMR 3.5, 95% CI 1.3-7.7) during pregnancy or within 42 days of pregnancy termination. The only other SMR that was > 1 was for death from cerebrovascular disorders during pregnancy and up to 42 days postpartum, although not significantly so (SMR 1.4, 95% CI 0.8-2.2). No other cause-specific SMRs were > 1. Moreover, recently pregnant women were found to be much less likely to commit suicide or to be the victims of homicide. We found no empirical justification for including deaths not directly related to pregnancy in reported counts of maternal deaths for most of the causal categories we considered. PMID- 11876836 TI - [Repairing severe deep burn wound with transplantation of the latissimus dorsi myocutaneous flap]. AB - OBJECTIVE: To repair the deep wound caused by high voltage electric injury and hot mangle injury with the latissimus dorsi myocutaneous flaps. METHODS: Latissimus dorsi myocutaneous flaps were applied to repair deep burn wounds in 26 cases in which 25 were island flap transposition and 1 free flap transplantation. RESULTS: The biggest flap in this group was 40 cm x 20 cm. All flaps survived but one exception with tip necrosis of the flap. Reconstruction of the Achilles tenden was satisfactory. CONCLUSION: Latissimus dorsi myocutaneous flaps are the ideal choice for repairing local severe deep burn wounds. PMID- 11876837 TI - [Mechanism of recombinant human growth hormone accelerating burn wound healing in burn patients]. AB - OBJECTIVE: To investigate the mechanism of recombinant human growth hormone (rhGH) in acceleration of burn wound healing. METHODS: Thirty three severe burn patients were enrolled in this study and randomly divided into rhGH group (n = 18) and placebo control group (n = 15). The rhGH was administered subcutaneously in the dose of 0.1 mg . kg(-1) at 10 pm every evening in rhGH group for 14 days, while normal saline was used as a placebo in the control group. The serum concentration of GH and IGF-1 were assayed with ELISA method, and the proliferation index (PI) and S-phase fraction (SPF) of wound tissue was analyzed by flow cytometry (FCM). The expression of PCNA, EGFR and CK was assessed by immunohistochemical stains. RESULTS: Serum concentrations of GH and IFG-1 increased significantly after rhGH administration. The PI and SPF of wound tissue were obviously higher than those before rhGH administration and those in the control group. The expressions of PCNA, EGFR and CK was enhanced. In addition, the epidermis of healed wound was thicker in rhGH group than that in the control group. CONCLUSION: The application of rhGH in burn patients could improve the serum concentration of GH and IFG-1, promote DNA synthesis and PCNA and EFGR expression of epithelium, so that re-epithelialization of the burn wound could be accelerated. PMID- 11876838 TI - [Effects of thrombin peptides on wound healing and proliferation and migration of normal human epidermal keratinocyte (NHEK)]. AB - OBJECTIVE: To define the effects of thrombin peptides on wound healing and NHEK proliferation and migration. METHODS: A wound model was made with four 1.5 cm circular full thickness dermal excisions on the back of each Sprague-Dawley rat. 0.1 microgram (40 microliter) TP508 was applied to each circular excisional wound in 9 rats, the other 9 received saline only. Wound area was calculated with JAVA Jandel and IMAGE PRO software. NHEK945 proliferation was assessed by MTT assay and direct cell count with a Coulter Counter. Cell migration was determined by 48 well Boyden Chamber. Cells migrated onto the lower surface of the filter were assessed by a Chemi Imager 4000 Image Analyzer and expressed as spot density. RESULTS: Wound area in rats treated with TP508 was 73.7% and 45.4% of saline control on day 7 and 14, respectively. NHEK945 proliferation was accelerated after adding thrombin and TP508. The spot density of migrated cells was 76.7 plus minus 13.8 in medium alone. After adding 1 microgram/ml of thrombin and 10 microgram/ml of TP508, the spot density was 104.4 plus minus 12.2 and 109.4 plus minus 14.6, respectively. CONCLUSION: Results of this study suggest that both thrombin and TP508 have significant actions on wound healing and NHEK proliferation and migration, which is important in wound repair. PMID- 11876839 TI - [The effect of escharectomy during burn shock stage on the expression of ICAM-1 and TNF-alpha mRNA of rat pulmonary tissue]. AB - OBJECTIVE: To study the rules of postburn expression of IGAM-1 and TNF-alpha and the rule of the change in the MPO activity in pulmonary tissue; to explore the influence of escharectomy on the changes in the above indices and to clarify the importance of escharectomy during shock stage. METHODS: One hundred and seventy six Wistar rats with 30% TBSA III degree back scald were used. RT-PCR was used in the examination of the expression of pulmonary tissue ICAM-1 and TNF-alpha mRNA and of the change in pulmonary MPO activity after escharectomy. RESULTS: The expressions of pulmonary ICAM-1 and TNF-alpha mRNA began to increase at 4 hour postburn and reached peak level at 12 and 24 hours postburn, respectively. Their expressions returned to near control level 96 hours postburn in rats undergone escharectomy during shock stage. On the contrary, they remained at a relative high level even on 7th postburn day in both non-operated rats and the rats receiving escharectomy 96 hours postburn. In addition, pulmonary tissue MPO activity fell to near control level in rats undergone escharectomy during shock stage, but it maintained a high level in rats in which escharectomy was not done during shock stage. CONCLUSION: These findings suggest that eschar could induce the production of endothelial adhesion molecules. Therefore escharectomy as early as possible is very important to prevent the expression and release of adhesion molecules and the development of SIRS. PMID- 11876840 TI - [Effect of escharectomy and skin grafting of minipigs during burn shock stage on postburn energy expenditure]. AB - OBJECTIVE: To determine the changes in resting energy expenditure (REE) of minipigs in normal and postburn stage, and the effect of escharectomy during burn shock stage on energy expenditure. METHODS: Swine model with 35% TBSA III degree flame burn was used. Eighteen male minipigs were randomly divided into three groups: control group, escharectomy during burn shock stage group and escharectomy after burn shock stage group. REEs were monitored by means of Cardiorespiratory Diagnostic Systems (Medical Graphics Corporation, USA). RESULTS: The levels of REE in escharectomy during burn shock stage group decreased early and were obviously lower than those in other two groups. CONCLUSION: These findings suggest that escharectomy and skin grafting during burn shock stage could ameliorate hypermetabolism in major burn patients. PMID- 11876841 TI - [Prolonging the survival of transplanted alloskin by transferring the CTLA(4)Ig locally with recombinant adenovirus vector in mice]. AB - OBJECTIVE: To find a simple and practical way to prolong the survival of transplanted alloskin to cover the burn and other wounds. METHODS: A recombinant CTLA(4)Ig-Adenovirus expression vector was constructed by means of homo recombination, and then transferred either into cultured murine skin grafts ex vivo or local wound surface. The expression of CTLA(4)Ig in cultured skin was immunohistochemically determined. Moreover, the survival time of transplanted alloskin from Kunming mice to Balb/c mice was evaluated. RESULTS: The CTLA(4)Ig Adenovirus vector could be successfully transferred and expressed in the cultured murine skin, and the locally used recombinant CTLA(4)Ig-Adenovirus vector could significantly prolong the survival of the transplanted alloskin from 8 d to 21 d. CONCLUSION: Local transfection of the CTLA(4)Ig-Adenovirus vector is a simple and effective way to prolong the survival of alloskin grafts. PMID- 11876842 TI - [Myocardial apoptosis induced by delayed fluid resuscitation in a burned rat model]. AB - OBJECTIVE: To explore the possibility and the mechanism of myocardial apoptosis induced by delayed fluid resuscitation in a burned rat model and its relationship with nitric oxide (NO) and oxygen-derived free radicals. METHODS: In a rat model with 30% III degree burn, the genomic DNA of the myocardial tissue was detected with ApoAlert(TMLM)-PCR Ladder Assay Kit and visualized with agarose gel electrophoresis. Meanwhile, the NO and the content of unsaturated fatty acids were measured. RESULTS: In rats receiving delayed fluid resuscitation following burn, the myocardial genomic DNA exhibited DNA ladder-index of apoptosis, and the contents of myocardial NO and unsaturated fatty acid were much lower than those in rats receiving immediate resuscitation (P < 0.05). CONCLUSION: The myocardial tissue undergoes apoptosis in burned rats receiving delayed fluid resuscitation, and the decreased NO and the production of oxygen-derived free radicals are also observed in this process. PMID- 11876844 TI - [The role of PMNs in early progressive injury of deep partial-thickness burn wound]. AB - OBJECTIVE: To examine the role of polymorphonuclear neutrophils (PMNs) in early progressive injury of deep partial-thickness burn wound. METHODS: After deep partial-thickness burn, the count of PMNs in beripheral circulation, activity of myeloperoxidase (MPO) and content of MDA in wound tissue of Spraque Dawley rats, were measured. Sections of wound biopsies were stained with Masson's thrichromatic staining and anti-vimentin immunohistopathologic staining for identifying the injured tissue. RESULTS: PMNs counts increased immediately and reached a peak at 4 h postburn, and MPO activity peaked at 24 h postburn. MDA content was lowe at 0.5 h and higher at 48 h postburn than that of normal control. Histologic analysis showed that there was more necrotic tissue at 48 hr postburn than that at 24 h. CONCLUSION: It was after 24 h postburn that PMNs induced tissue damage. The results suggest that intervention of PMNs adhesion should be under taken in about 4 h after thermal injury and anti-oxidizing agent should be given before 24 h postburn in order to prevent early progressive injury to the wound. PMID- 11876845 TI - [Effects of cytokine gene therapy on prolonging survival time of allografted skin after scalding in a murine model]. AB - OBJECTIVE: To explore the role of cytokine gene therapy on prolonging survival time of allografted skin after scalding in a murine model. METHODS: Interleukin 10 (IL-10) gene was employed as therapeutic objective gene and fibroblast was as a carrier cell. Gene transcription technique was adopted to establish an experimental murine model in which fibroblast-mediated gene therapy was used to prolong allografted skin survival time after scalding. IL-10 was transferred into fibroblastocyte (NIH3T3) by reverse transcriptive virus vector. The mice were grafted with alloskin after scalding. In addition, collagen capsulized NIH3T-3-IL 10 cells were implanted intraperitoneally in the mice so as to observe its influence on allografted skin survival time and on the changes of their main internal organs. RESULTS: Cytokine gene therapy could obviously prolong the survival time of allografted skin (P < 0.01) without any evident detrimental effect on the internal organs. CONCLUSION: These results demonstrated that skin allograft rejection could be inhibited and the survival time be prolonged with the implantation of the fibroblastocyte transferred IL-10 gene. PMID- 11876843 TI - [The change in hyaluronic acid (HA) in auto-and allo-skin intermingled grafting in extensive full thickness burn in rat model]. AB - OBJECTIVE: To elucidate the changes in hyaluronic acid (HA) contents in auto-and alloskin intermingled grafting. METHODS: HA was determined by radio-immunoassay in auto-and alloskin intermingled grafting (a large sheet of allograft inlaid with small stamps of autoskin) in experimental extensive full-thickness burn rats on 3rd, 5th, 7th, 14th, 21st, 28th postoperation days. RESULTS: In both areas of intermingled skin grafting, the contents of HA was reduced first and increased significantly thereafter, but the contents in auto-skin areas rose earlier and higher than those in allo-ones. CONCLUSION: The results suggest that adequate application of HA may be beneficial to the healing and remodelling of grafted area. PMID- 11876846 TI - [Effects of Asiaticoside on hypertrophic scars in a nude mice model]. AB - OBJECTIVE: To study the effect of Asiaticoside on the fibroblast collagen biosynthesis in vitro culture and on the hypertrophic scar in a nude mice model, and its mechanism. METHODS: Light microscope (LM) and electron microscope (EM) were employed to study the morphological changes in fibroblast before and after the application of Asiaticoside. The fibroblast collagen synthesis was assayed by (3)H-proline incorporation. A nude mice model with hypertrophic scar was established for observing the effect of Asiaticoside given by local injection. RESULTS: It was found that Asiaticoside could significantly affect the ultrastructure of fibroblast and inhibit its collagen synthesis in vitro culture (P < 0.01) in a manner of dose-effect relationship. Local injection of Asiaticoside into the nude mouse body could inhibit the proliferation of scar without any toxic effect. CONCLUSION: The possible mechanism of the effect of Asiaticoside on hypertrophic scars is related to its inhibitory action on the fibroblast proliferation and collagen synthesis. PMID- 11876848 TI - [The change of intestinal microcirculation blood flow and its relationship with bacterial translocation in burned rats receiving delayed fluid resuscitation]. AB - OBJECTIVE: To investigate the change of intestinal microcirculation blood flow and its relationship with bacterial translocation in burned rats receiving delayed fluid resuscitation. METHODS: Twenty-four Wistar rats inflicted by 30% TBSA III degree followed by delayed fluid resuscitation were taken as the model. The rats were randomly divided into four groups, i.e. burn shock without resuscitation (BSNR), delayed resuscitation (fluid given from 8 hours postburn, DR), immediate resuscitation (IR), and sham operation (SO). The incidence of bacterial translocation, intestinal microcirculation blood flow volume of intestinal wall at terminal ileum as well as the change of systemic mean arterial pressure were monitored. RESULTS: The incidence of bacterial translocation in DR group (54.2%) was significantly higher than that in IR group (P < 0.01), but no remarkable difference with that in BS group (P < 0.05). In addition, microcirculation blood flow of intestinal wall recovered to some degree at 4 hours after delayed fluid resuscitation but far below that in IR group at the same time point. Nevertheless, the systemic mean arterial blood pressure had restored to the normal range at that time. CONCLUSION: The incidence of intestinal bacterial translocation sustained at high level after burn shock with delayed fluid resuscitation, which might be the result of delayed improvement of microcirculation blood flow of the intestinal wall. PMID- 11876847 TI - [The experience of the management of burn sepsis with different strategies in our department during the past 29 years]. AB - OBJECTIVE: To retrospectively sum up the experiences in the prevention and treatment of sepsis after thermal injury. METHODS: From January 1970 through October 1998, altogether 5 330 burn patients were admitted to our burn center, and among them 451 patients developed sepsis. To analyze the efficacy of different treatment strategies developed during these 29 years, three periods were divided, namely 1970 to 1979, 1980 to 1992, and 1993 to 1998. The incidence and mortality of sepsis were compared, thereupon the efficacy of different treatment strategies were analyzed. RESULTS: The overall incidence and mortality of sepsis in all patients and those in patients with burn extent exceeding 30% TBSA were significantly lower in the latter period compared with the former two periods (P < 0.05 similar 0.01). In addition, in the last period, blood levels of LPS, TNFalpha, IL -- 6, and IL -- 8 were obviously higher throughout the course of MODS, especially before death, in the patients who finally died of sepsis, complicated by MODS, compared with those in the survivors (P < 0.05 similar 0.01). CONCLUSION: Summing up our experiences, it is our belief that the treatment strategies for prevention of postburn sepsis in extensively burned patients should include rapid, adequate fluid resuscitation for burn shock, effective control of infection, early excision of deep burnwounds followed by good skin coverage, and reinforcement of organ support. Once burn wound sepsis occurs, prompt removal of infected necrotic tissue is the key procedure to ensure a successful result. PMID- 11876849 TI - [The sifnificance and characteristics of the internal distribution of Staphylococcal enterotoxin B in burned rats with sepsis]. AB - OBJECTIVE: To investigate the role of staphylococcal enterotoxin B (SEB) in the development of postburn staphylococcus aureus sepsis. METHODS: Rats inflicted with 20% TBSA III degree scald followed by Staphylococcus aureus sepsis were used as the experimental subjects. The distribution of SEB in the vital organs such as the heart, liver, lungs, kidneys and plasma was observed with concomitant serial measurement of the relative organ functional indices. RESULTS: The plasma SEB content increased transiently in the early stage of sepsis (P < 0.01) and then fell to near that of control group. The SEB contents in the heart, liver, lungs and kidneys increased evidently 2 hours after staphylococcus aureus challenging (P < 0.01) and continued to increase afterwards. The functions of multiple organs were impaired obviously at 24 postburn hours, and the function impairment was aggravated by S. aureus challenge. The degree of organ injury of liver and kidneys correlated positively with tissue content of SEB (P < 0.05 similar 0.01). CONCLUSIONS: Staphylococcal enterotoxin B might play a role in the development of MODS induced by postburn sepsis. PMID- 11876850 TI - [The significance and the role of TNFalpha and NO in the early renal damage in burned rats complicated with endotoxemia]. AB - OBJECTIVE: To explore the role and the significance of TNFalpha and NO in the early renal damage in burned rats complicated with endotoxemia. METHODS: The rats inflicted with 20% TBSA III degree burn and injected intraperitoneally with endotoxin (1 mg/kg) were employed as the model of MOD. The renal pathomorphology was observed at 0.5, 1, 3, 6, 12, 24, 48, 72 postburn hours. Concomitantly, the changes of renal and serum TNFalpha and NO were investigated and in situ hybridization for TNFalpha mRNA and immunohistochemical staining for iNOS were performed. RESULT: The expressions of renal iNOS and TNFalpha mRNA, the NO synthesis and the serum levels of NO and TNFalpha in BCEG (burns complicated with endotoxin group) were obviously higher and appeared earlier than those in SBG (simple burns group) and SEG(simple endotoxin group). CONCLUSION: The renal TNFalpha and NO increased significantly in rats subjected to burn complicated with endotoxemia, which might be important factors causing renal damage and the alterations in renal hemodynamics. PMID- 11876851 TI - [An experimental study of antibiotic -- induced endotoxin release in Pseudomonas aeruginosa bacteremia inburned rats]. AB - OBJECTIVE: To assess the intensity of endotoxin release in Pseudomonas aeruginosa challenged burned rats after the administration of various antibiotics. METHODS: Rats were inflicted with 30% TBSA burn, and they were challenged with Pseudomonas aeruginosa (PA103). After the administration of different beta lactam antibiotics, serum endotoxin level and blood bacterial count were determined. RESULTS: All the antibiotics used were effective in eradicating the bacteria with concomitant release of endotoxin in varying amounts. It was found that Imipenem released the least amount of endotoxin, followed in order by Cefoperazone, Ceftazidime, and Cefotxime. CONCLUSIONS: Different antibiotics possessed varying capacity of inducing endotoxin release from PA, and the amount released did not bear any relation with the bactericidal capability of the antibiotics. PMID- 11876852 TI - [Increased expression of peritoneal macrophage CD14 in severely burned mice]. AB - OBJECTIVE: To observe the dynamic changes in peritoneal macrophage (Mphi) CD14 expression in severely burned mice. METHODS: The mice, being inflicted with 20% TBSA III degree burn, were randomly divided into two groups: Group A: The mice in this group received fluid resuscitation instantaneously after burn. Group B: The mice in this group received delayed fluid resuscitation at 6hr postburn. The Mphis at different postburn time points were obtained in two groups for the detection of their expression of CD14 and CD14 mRNA with immunohistochemical and RT -- PCR analysis, and also for the determination with ELISA of the changes in TNF and IL -- 6 levels in the supernatant of their culture. RESULTS: The expression of CD14 and the levels of TNF and IL -- 6 increased obviously in both groups from 1 to 24 hrs postburn. CONCLUSION: Burn injury would induce the increase of the expression of Mphi CD14 mRNA in mice. As a result, the Mphi became more readily to be activated by low levels of LPS. The activated Mphi might play an important role in the development of postburn SIRS. PMID- 11876853 TI - [Effect of serum and wound exudates on PMN apoptosis in burned patients]. AB - OBJECTIVE: To study in vitro the effect of burn serum and wound exudates on PMN apoptosis in burned patients. METHODS: The burn serum was obtained from patients within 3 days postburn. The blister fluid was collected from intact blisters in burn wound within 24 hours postburn, and subeschar tissue fluid underneath III degree burn wound during escharectomy. PMNs were isolated from healthy human donors and cultured for 24 hours at 37 degreeC and 5% CO(2) with normal serum (NS), burn serum (BS), burn blister fluid (BBF) and subeschar tissue fluid (STF), respectively. Morphological assessment and quantification of PMN apoptosis were performed with AO + EB stain and flow cytometric analysis. RESULTS: The number of apoptotic PMN and DNA fragmentation of PMN significantly decreased (P < 0.05) when PMN were cultured with BS, BBF, and STF when compared with that with NS. In addition, the specific DNA "ladder" pattern for apoptotic cells couldn't be found on the agarose gel electrophoresis in this case. However, BBF exhibited weaker inhibiting effect than BS and STF on PMN apoptosis. CONCLUSION: PMN apoptosis could be inhibited in vitro by BS, STF and BBF from burned patients. PMID- 11876854 TI - [Study on the expression of integrin alpha(5), beta(1) during wound healing]. AB - OBJECTIVE: To explore the potential role of integrin family in mediating the interaction between cells and extracellular matrix (ECM) during wound healing. METHODS: In situ hybridization technique was employed in the determination of the expression of the mRNA of the subunits of integrin alpha(5), beta(1) on the surface of fibroblasts in the donor sites of partial thickness skin of 12 burned patients. RESULTS: Evidently more integrin alpha(5), beta(1) positive staining fibroblasts were found in the donor site than in normal skin (P < 0.05). CONCLUSION: The results suggested that increased expression of integrin alpha(5), beta(1) in the wound might be involved in the activation of collagen-- overproductive subpopulation of fibroblasts. PMID- 11876855 TI - [The clinical significance of the change of blood testosterone in burned patients]. AB - OBJECTIVE: To investigate the patterns and significance of the change in blood testosterone in burned patients. METHODS: The changes of blood testosterone and luteinizing hormone (LH) were dynamically monitored in 21 male patients with moderate burn injury. RESULTS: The blood testosterone levels in this group decreased persistently after burn, and the decrement degree was related to the burn severity. But LH exhibited no regular change and its change exerted no effects on the blood testosterone levels. CONCLUSION: Decrease in blood testosterone in burned patients was related to gonadal injury, which caused the shortage and insufficiency of anabolic hormone and also of protein synthesis. The correction of the shortage of testosterone might be beneficial to protein synthesis and to the restoration of positive nitrogen equilibrium in burned patients. PMID- 11876857 TI - [The effect of burn sera on the calcium homeostasis in isolated rat myocardial cells]. AB - OBJECTIVE: To study the effect of burn sera on the calcium homeostasis in isolated rat myocardial cells. METHODS: The burn sera were obtained from the Sprague -- Dawley (SD) rats inflicted by 30% TBSA III degree burn at 2, 4 and 6 postburn hours with the harvesting of burn sera at different postburn times, named as 2 hBS, 4hBS and 6 hBS, respectively. Myocardial cells were isolated by enzyme digestion with retrograde perfusion apparatus. [Ca(2+)]i was detected with the fluorescent dye Fura -- 2 and L -- type calcium (ICa -- L) currents were recorded in the whole -- cell patch -- clamp model. RESULTS: Myocardial cellular [Ca(2+)]i became much higher in concent after being in cubated with sera, especially with 6hBS, than that in normal myocardial cells (P < 0.01). But the effect of 6hBS could be significantly inhibited by verapamil (30 nmol/L), the calcium channel antagonist and procaine (2 mmol/L), the inhibitor of rynodine receptor (P < 0.01) by 47.7% and 67.6%, respectively. Ca -- L was evidently increased by the stimulation of 2 hBS and 6 hBS, by 50 -- 80% and 1.5 -- 2.5 folds, respectively. It was indicated by I -- V curves that every ICa caused by depolarizaton and clamp voltage was increased by burn sera, which further made the maximal activating voltage ahead of time. All the burn sera effects on calcium current could be removed by extracellular lavage. CONCLUSION: The myocardial [Ca(2+)]i and ICa -- L could increase significantly when affected by burn sera. The alteration in calcium homeostasis might be a cellular mechanism of myocardial injury. PMID- 11876856 TI - [The relationship between postburn gene expression of modulators in gut associated lymph tissue and the change in IgA plasma cells]. AB - OBJECTIVE: To investigate the relationship between the postburn change in IgA plasma cells and those of IL- 4 and IL-6 in gut associated lymph tissue (GALT). METHODS: One hundred and twenty -- five SPF mice were enrolled in the study and randomly divided into three groups -- normal control (A), microbial inoculation (B) and microbial inoculation followed by burn injury. (C) In groups Band C, the mice were gavaged with candida albicans, and the mice in group C, were inflicted with 20% TBSA III degree burn at the 14th day after inoculation. The mice were sacrificed thereafter on 1, 2 and 3 postburn days and the samples were obtained aseptically. The number of candida albicans adhering to intestinal mucosal membrane was counted. The number of IgA plasma cells in lamina propria was determined by immunohistochemical staining. The gene expression of IL-4 in Peyer patch was detected by dothybridization and that of IL-6 in lamina propria was determined by insitu hybridization. RESULTS: (1) The postburn gene expression of IL-4 in Peyer patch decreased, but the IL-6 level in intestinal lamina propria increased. (2) The number of IgA plasma cells in lamina propria decreased significantly, but the adherence of candida albicans to intestinal mucous membrane increased obviously. CONCLUSION: The decrement of postburn gene expression of IL-4 in Peyer patch might lead to IgA plasma cell decrease, resulting in the increase of candida albicans adhering to intestinal mucosal membrane. PMID- 11876858 TI - [Treatment of severe inhalation injury with endotracheal infusion of exogenous pulmonary surfactant]. AB - OBJECTIVE: To explore the possibility and the effects of clinical application of exogenous pulmonary surfactant (PS). METHODS: Two burned patients with severe inhalation injury were treated with 5% emulsified porcine PS by endotracheal infusion of 10 ml every 2 hours with the assistance of mechanical ventilation. The indices of patients' oxygenation and related pulmonary dynamics were observed. RESULTS: Twenty-four hours after the application of exogenous PS, PaO(2) and PaO(2)/FiO(2) increased evidently, but (A -- a) DO(2) and RI decreased obviously and PaCO(2) sustained for the observing period. The index related pulmonary dynamics, DeltaV/DeltaP increased gradually. CONCLUSION: With the assistance of mechanical ventilation, endotracheal infusion of exogenous PS could improve oxygenation and compliance of patients with severe inhalation injury. And the patients exhibited very good tolerance to exogenous PS. PMID- 11876859 TI - [The clinical significance of the dynamic change in serum cardiac troponin T in burned patients]. AB - OBJECTIVE: To evaluate the diagnostic value of serum cardiac troponin T (cTnT) in myocardial injury in burned patients. METHODS: Sixty-two burned patients were cardiac divided into 3 groups, i.e. mild and moderate (M, n = 25), severe (S, n = 22) and extra (E, n = 15) groups on the basis of patients' severity. Radio immunoassay was adopted to detect serum cTnT and CK-MB in patients of all groups on 1, 3, 7 and 14 postburn day. Ultrasound cardiogram and ECG (electrocardiogram) were employed to monitor cardiac function. Twenty healthy volunteers were enrolled in the control group (C). RESULTS: (1). The serum cTnT levels at all time points in patients in S and E groups were evidently higher than those in C group (P < 0.01) and paralleled very well with the severity of the patients' injuries. (2). CK-MB exhibited similar change as that of cTnT. (3). There exhibited abnormal findings with ultrasound cardiogram and ECG in five cases in E group. CONCLUSION: The dynamic changes in serum cTnT and CK -- MB could well reflect the degree of the myocardial injury in burned patients, and cTnT was much more reliable as an index of myocardial injury than the serum enzymes due to its higher specificity and sensitivity. PMID- 11876860 TI - [The inhibitory effects of chloroquine and dexamethasone on the formation of pulmonary edema in the early stage of smoke inhalation injury in rats]. AB - OBJECTIVE: To explore the inhibitory effects of chloroquine and dexamethasone on the formation of pulmonary edema in the early stage of smoke inhalation injury in rats. METHODS: Wistar rats were used and randomly divided into 10 groups, i.e. normal control, 1 PBH (postburn hour), 3 PBH, 6 PBH, 12 PBH and 24 PBH groups of smoke inhalation injury, 6 PBH and 12 PBH groups with dexamethasone treatment, and 6 PBH and 12 PBH groups with chloroquine treatment. The lung water content (LWC), pulmonary vascular permeability (PVP) and hemorrhagic amount (PHA) were measured. RESULTS: The LWC and PVP (measured by the pulmonary exudation amount of (131)I -- labelled albumin) increased obviously, peaked at 6 PBH and lasted to 24 PBH (P < 0.01) in rats injured by smoke inhalation. In groups treated with chloroquine or dexamethasone, the LWC and PVP were significantly lower at 6 PBH (P < 0.05). Similarly, the PHA in injured group exhibited remarkable, especially at 1 PBH and up to 24 PBH when the value was still higher than that in normal control group (P < 0.01). In groups treated with chloroquine or dexamethasone, PHA abated evidently at 6 PBH (P < 0.01). But there were no significant difference of the indices among groups at 12 PBH. CONCLUSION: Both chloroquine and dexamethasone be effective in preventing and treating pulmonary edema in rats in the early stage of smoke inhalation injury. PMID- 11876861 TI - [A canine model of brain edema established in the early postburn stage]. AB - OBJECTIVE: To establish a canine model of brain edema in the early stage of severe burn. METHODS: Mongrel dogs were inflicted by napalm with 50% TBSA III degree burn. The dogs were randomized into normal control (C), burn (B), burn with balanced salt solution (S) and burn with glucose (G) groups. The macropathological, histopathological and ultrastructural changes of brain tissue and brain water (%) were examined at 6, 12, 18 and 24 postburn hours (PBH), respectively, with simultaneous evaluation of blood brain barrier by (99) TC -- ECD. RESULTS: Pathological evidence of brain edema exhibited at as early as 6 PBH in B, S and G groups, which included cellular swelling of partial endothelia and pericapillary astrocytes, and ischemic necrosis of endothelia, neurons and axons to varying degrees. All the changes exhibited more and more obvious with the elapse of postburn time. The brain water content in each burn group, especially in G group at 24 PBH, was more than that in C group. It was indicated by nuclide imaging that there appeared gradual increasing of the concentration of (99) TC -- ECD in brain tissue in B and C groups since 6 PBH. CONCLUSION: This model was established for the convenience of burn scholars to study the pathogenesis and management of postburn brain edema. PMID- 11876862 TI - [The significance of the expressions of lipopolysaccharide binding protein mRNA and lipopolysaccharide receptor CD14 mRNA in the liver of burned rat]. AB - OBJECTIVE: To explore the significance and the correlation between enteric endotoxin translocation and hepatic endotoxin -- sensitivity -- enhancing system- lipopolysaccharide -- binding protein (LBP)/lipopolysaccharide receptor CD14 (LBP/CD14) after burn. METHODS: Wistar rats subjected to 35% III degree burn were employed as the model. The rats were randomly divided into three groups: i.e. normal control (C, n = 8) group, thermal injury (T, n = 10) group and recombinant bactericidal/permeability -- increasing protein (rBPI(21)) treatment (R, n = 6) group. The rats in T and R groups were sacrificed at 12 postburn hour (PBH) and the liver tissue was collected for the detection of the mRNA expressions of LBP, CD14 and tumor necrosis factor (TNFalpha) and blood samples collected for hepatic functional indices. RESULTS: The hepatic endotoxin content increased significantly postburn (P < 0.01), and the expressions of LBP/CD14 and TNFalpha mRNA in liver tissue increased obviously. However the use of rBPI(21) could evidently lower hepatic endotoxin content and inhibit the expression of tissue LBP/CD14 and TNFalpha. In addition, rBPI(21) could also significantly decrease serum level of glutamic -- pyruvic transaminase (ALT) (P < 0.01). CONCLUSION: Aggregation of endotoxin in liver due to postburn translocation might obviously stimulate the expression of LBP/CD14 mRNA locally. The up -- regulation of the expression of LBP/CD14 might be the principle molecular basis enhandced activity of translocated endotoxin on inflammatory cells. PMID- 11876863 TI - [A report of 115 cases of amputation after electric injury]. AB - OBJECTIVE: To analyze the main features of amputation subsequent to electric injury. METHODS: One hundred and fifteen cases of amputation were analyzed with the purpose to define the indication, the optimal level, operational methods and the prognosis. RESULTS: The incidences of amputation in this group were 45.4% for upper and 14.9% for lower extremities, respectively. The indications were total necrosis of the extremities, secondary necrosis of the extremities due to the thrombosis, bleeding and rupture of the major supplying arteries and failure to restore severely injured tissue. The commonest amputation levels were the mid third and the upper third of the forearms. The primary healing rate was 63.5%. CONCLUSION: It was very important to identify the optimal indications, the proper levels and operational methods for the amputation after electric injury, so as to make stumps more suitable for the installation of artificial limbs to restore better function. PMID- 11876864 TI - [The role of ozone solution on debridement and sterilization of burn wound]. AB - OBJECTIVE: To observe the role of ozone solution on debridement and sterilization of burn wound. METHODS: In vitro sterilizing effect on common isolated bacteria from burn wound and debridement and sterilization effects on burn wound of ozone disinfectant (ozone solution) were studied. RESULTS: All the bacteria tested were killed in vitro by ozone solution. In addition, when ozone solution was applied on burn wound, its clearance rate of bacteria was 94.5% and the clinical effective rate was 97.1%. CONCLUSION: Ozone is low in cost and high in effect which might be used as an agent for burn wound disinfection. PMID- 11876865 TI - [The effects of massive escharectomy on the resuscitation of burn shock in the early stage after burn]. AB - OBJECTIVE: To evaluate the effects of massive escharectomy on the resuscitation of burn shock in the early stage after severe burn. METHODS: Mongrel dog inflicted with 40% TBSA III degree scalding were employed as the model. The dogs were randomly divided into two groups ------ escharectomy (E) and non -- escharectomy (C) groups. All the dogs received balanced salt solution according to Parkland formula from 1 postburn hour (PBH). The dogs in E group underwent escharectomy en masse at 3 PBH with immediate grafting of fresh alloskin. The dynamic indices of hemodynamics, blood rheology, oxygen supply, oxygen consumption and blood lactic acid content were monitored. RESULTS: Severe shock occurred in all the dogs 30 min after burn. The were evident decreased cardiac output (CO), cardiac index (CI) and left ventricular work index (LVWI) in all the dogs. But the viscosity of whole blood and plasma, the erythrocytic agglutination index and plasma lactic acid content increased significantly. The dogs in E group were operated with massive escharectomy at 3 PBH. In this group, CO, CI, blood rheologic indices and plasma lactic acid content were obviously improved when compared with those in the preoperational period and to those in the C group. CONCLUSION: Simple fluid resuscitation couldn't correct burn shock quickly. Massive escharectomy during shock stage was safe, feasible and effective in the correction of burn shock. PMID- 11876866 TI - [Clinical analysis of 106 cases of inhalation injury induced by gas explosion]. AB - OBJECTIVE: To review the management of inhalation injury induced by gas explosion in our department, so as to draw some experience and lessons for future work. METHODS: One hundred and six cases of inhalation injury induced by gas explosion were analyzed in terms of the characteristics of and therapeutic methods for the disease. RESULTS: The injury induced by gas explosion was very complicated with high incidences of compound injuries, severe inhalation injury and systemic infection. In addition, respiratory dysfunction occurred very quickly and severely at early postburn stage, which could easily develop to respiratory failure. The latter could ultimately lead patients to death due to improper management. CONCLUSION: In order to raise cure rate of the inhalation injury induced by gas explosion, airway obstruction should be cleared immediately and fiberbronchoscopy and respirator should be employed as soon as possible to help remove endotracheal obstruction and to maintain effective gas exchange. In addition, correct pulmonary resuscitation and continuous blood gas analysis were advocated. PMID- 11876867 TI - [The effects of NAC on the expression and activity of SPA in rats inflicted by smoke inhalation injury]. AB - OBJECTIVE: To investigate the effects of NAC (N-acetyl-L-cysteine) on the expression and activity of SPA (surfactant-associated protein A) in rats inflicted by smoke inhalation injury. METHODS: Wistar rats inflicted with smoke inhalation injury were employed as the model. The expression of SPA mRNA, the static pulmonary compliance, the surface tension of the alveolar lavage and the morphology of lamellae bodies (LB) of type II alveolar cells were examined. RESULTS: After the application of NAC, there exhibited an increase in SPA mRNA expression and static pulmonary compliance, and the restoration of the BAL surface tension to normal. CONCLUSION: The application of NAC could promote the expression of SPA mRNA after smoke inhalation injury and improve the SPA-related function. PMID- 11876868 TI - [The effects of hypertonic saline solution perfusion on the isolated hearts of burned rats]. AB - OBJECTIVE: To observe the effect of hypertonic saline solution perfusion on the function of isolated hearts of burned rats. METHODS: The isolated rat heart was taken as the model. The hearts from normal control and burned rats were perfused by improved isotonic (Na(+) 118 mmol/L) and hypertonic Krebs-Henseleit (Na(+) 215 mmol/L) solutions, respectively. The dynamic changes of the cardiac function were monitored during heart perfusion. RESULTS: The cardiac contractive and diastolic function were transiently inhibited and followed by quick recovery during the perfusion with hypertonic solution in both control and burned rat hearts. But when the hypertonic solution was replaced by isotonic, there appeared significant enhancement of the cardiac contractive and diastolic function. These findings indicated that hypertonic solution exerted more powerful influence on the contractive and diastolic function of burned rat heart. CONCLUSION: Hypertonic saline solution could influence the cardiac contractile and diastolic function significantly. And the magnitude of the effects was correlated with the basic cardiac condition. PMID- 11876869 TI - [The expressions of transcription factor c-fos mRNA and its protein in the burn wound of rats]. AB - OBJECTIVE: To investigate the rule of the expression in different time points of wound tissue transcription factor c-fos mRNA and its protein after burn, so as to explore the role of c-fos in the initiation of burn wound injury and repairing. METHODS: Male Wistar rats were inflicted with partial thickness scalding of 30% TBSA on the back. In situ hybridization and immunohistochemical methods were used to evaluate the characteristics and pattern of the changes in the expressions of the c-fos mRNA and its protein in normal skin and in the wounded skin at 3 h, 6 h, 1 d, 3 d, 7 d and 14 d postburn, respectively. RESULTS: It was found by light microscopy (LM) that c-fos mRNA and its protein could be found expressed in normal skin and obviously increasingly expressed in wounded skin. The expression of fos protein reached peak level at 3 postburn hour (PBH) and that of its mRNA at 6 PBH. CONCLUSION: The expression of c-fos mRNA and its protein could be induced during burn injury. But the postburn increase in the expression of its gene appeared later than that of its protein, suggesting that the early-appeared fos protein acted through its post-translational modification of preexisting fos molecule. PMID- 11876870 TI - [The effects of zinc supplementation on postburn nutritional status of zinc deficient scalded rats]. AB - OBJECTIVE: To investigate the effects of zinc supplementation on postburn nutritional status of zinc-deficient scalded rats. METHODS: Weanling Wistar rats were fed with zinc-deficient diets for one week to induce zinc-deficient status. The rats were inflicted with 20% TBSA deep partial thickness burn and thereafter fed with diets of low, normal and high zinc contents. The scalded rats fed with normal zinc-containing diet before and after injury were taken as normal control group. The changes of body weight, cumulative nitrogen balance and the contents of DNA and protein in jejunum mucous, liver, kidneys, spleen and testis were employed to represent the nutritional status of the rats. RESULTS: The rats fed with diets with normal and high contents of zinc were superior to those with low content of zinc in terms of body weight, food conversion rate, cumulative nitrogen balance and organ protein contents. The serum zinc level, alkaline phosphatase (ALP) activity and the DNA contents in jejunum mucous, kidneys, spleen and testis increased significantly after zinc supplementation. CONCLUSION: Zinc supplementation was helpful to improve postburn nutritional status in scalded rats. And relative higher amount of zinc supplementation might be beneficial to raise the DNA content in testis and kidneys when compared to that with normal zinc supplement. PMID- 11876872 TI - [Alterations of anabolism in hepatic cells of burned rats measured by (13)C NMR spectroscopy]. AB - OBJECTIVE: To introduce a safe and specific approach of (13)C NMR spectroscopy and investigate the alterations in hepatic anabolism functions including pyruvate recycling, gluconeogenesis and anaplerosis in severely burned rats. METHODS: Adult Spreque -- Dawley (SD) rats were catheterized arteriovenously via left carotid and right external jugular vein under inhalation anesthesia and were randomly divided into burn group (B) and sham burn group (S). Fluid resuscitation started 20 min postburn. Sodium [1, 2, 3 -- (13)C] propionate (SP) was intravenously administered as a tracer at 72 postburn hour (PBH). (13)C NMR spectroscopy of extracted plasma and liver, arterial blood pressure, heart rate and plasma glucose and sodium levels were measured. Metabolic parameters were calculated using a mathematical model of input -- output matrix to feed the tricarboxylic acid cycle. RESULTS: When compared to those in C group, burned rats exhibited a lower arterial blood pressure, but normal blood glucose at 72 PBH. Intravenous SP infusion did not alter MAP, heart rate and plasma sodium. The form of blood sugar labeled by (13)C isotopomer was in excellent agreement with that from liver. After continuous infusion of SP for 45 mins, SP could reach metabolic steady status. The hepatic cellular gluconeogenesis, anaplerosis and the carbon flow rate in pyruvate recycling significantly increased in B group when comparing to those in C group. But the carbon flow rate in Kreb's cycle decreased relatively, and hepatic pools of alanine, glutamic acid and glutamine were enlarged obviously in burned rats relative to sham burn group. CONCLUSION: These data suggested that intravenous infusion of SP would not alter MAP, heart rate and blood sodium. Forty -- five mins after the intravenous infusion, the tracer could reach metabolic steady status. The form of blood sugar labeled by (13)C could represent that of hepatic sugar. Seventy -- two hours after major burns, there were increased hepatic anaplerosis, gluconeogenesis and pyruvate recycling and a correspondent increased utilization ratio of blood sugar by peripheral tissue. PMID- 11876871 TI - [Effects of early enteral feeding and L-arginine supplementation on liver albumin synthesis in burned rats]. AB - OBJECTIVE: To evaluate the effect of early enteral feeding and L-arginine supplementation on mRNA expression of albumin, and some cytokines. METHODS: Twenty Wistar rats prepared with gastrostomy 7 days before the experiment were inflicted with 30% TBSA flame and were divided into early enteral feeding group (EF) and L-arginine enriched early enteral feeding group (AEF). All rats in two group starts enteral feeding two hours after burn. Capillary electrophoresis and reverse transcription-polymerase chain reaction (RT-PCR) were used to measure the mRNA of albumin, TNFa, IL-1a, IL-6R and iNOS respectively on 1st, 3rd, 6th and 9th day post burn (PBD). Serum albumin was observed at the same time. RESULTS: The mRNA expression of albumin on PBD1 was reduced in both group and improved on PBD3, 6, 9. The gene expression of albumin was significantly higher in AEF group than in EF group onPBD3, 9 [(1812 +/- 41) vs (1417 plus minus 43), P < 0.01]. Similarly, serum albumin were very low in two groups on PBD1 and improved on PBD3, 6, 9. Serum albumin in AEF group was significantly higher than in EF group on PBD9 [(31.9 plus minus 2.1) g/L vs (28.1 plus minus 1.3) g/L, P < 0.01]. The mRNA of TNFalpha, IL-1alpha, IL-6R and iNOS were increased on PBD1 and reduced on PBD3, 6, 9. The gene expression of TNFalpha, IL-1alpha in AEF group were significantly lower than EF group on PBD6, while IL-6R differed significantly between AEF group and EF group [(972 +/- 87) vs (2570 +/- 90), P < 0.01] on PBD9. There were no significant difference in iNOS mRNA production after burn in two group. CONCLUSION: Early enteral feeding could improve albumin biosynthesis and reduce the gene expression of TNFalpha, IL-1alpha, IL-6R and iNOS. L-arginine supplemented early enteral feeding could further increase the gene expression of albumin possibly by reducing the expression of TNFalpha, IL-1alpha and IL-6R and did not increase the expression of iNOS. PMID- 11876873 TI - [Effects of different nutritional support routes on the intestinal mucosa mucosainjury and renovation in burned rats]. AB - OBJECTIVE: To compare the effects of different nutritional support routes on the intestinal mucosa in burned rats. METHODS: Wistar rats inflicted with 30% TBSA full thickness burns were employed as the model and randomly divided into three groups: burn with enteral nutrition (EN), burn with parenteral nutrition (PN), and normal control (C). Solutions for intravenous and oral application with same volume, calorie, nitrogen and other nutrients were administered to rats in PN and EN groups, respectively. The plasma diamine oxidase (DAO) activity, the intestinal mucosa change of proliferating cell nuclear antigen (PCNA), of transmembrane electric potential difference (PD), of intestinal trefoil factor and their correlation analysis (ITF) were observed. RESULTS: Plasma DAO activity increased obviously and PD, PCNA values, ITF content were much lower in EN, PN groups than those in C group. The ITF content, the values of PD and PCNA in EN were much higher than those in PN group, while DAO activity in EN group was obviously lower than that in PN group. In addition, ITF content was significantly negatively correlated to plasma DAO activity, and obviously positively correlated to PCNA and PD values. It is suggested that intestinal mucosa injury was much more severe in PN group than that in EN group. CONCLUSION: Postburn intestine mucosa mucosal structural injury was related to the evident decrease of synthesis and secretion of ITF. Enteral nutrition might slow down the decrement of ITF, which could explain why EN was superior to PN in terms of decreasing intestinal mucosal injury and enhancing intestinal renovation. PMID- 11876874 TI - [Analysis of the factors related to the change of energy expenditure in burned patients]. AB - OBJECTIVE: To analyze the factors related to the change of energy expenditure in burned patients. METHODS: Indirect calorimetry was employed to detect energy expenditure (EE) dynamically in 45 cases of burned patients and to analyze their change. RESULTS: EE increased immediately postburn and reached top level in about one week (11, 443.78 plus minus 3, 295.32) kJ/d. EE value was positively correlated to the size of burn area. EE was also correlated to the development of the disease. The EE variation happened in the early postburn stage. The degree of variation was positively correlated to the size of burn area. The EE value in patients with medium size burn and inhalation injury was higher than that without inhalation injury. Postoperative EE decreased obviously in patients with major burn who had undergone escharectomy. EE in male patients was higher than that in female. CONCLUSION: Postburn EE varied with some regularity, but was affected by burn depth and size, postburn time, inhalation injury, escharectomy, sex and individual difference. PMID- 11876875 TI - [Analysis of riboflavin status in severely burned patients]. AB - OBJECTIVE: To investigate riboflavin status in severely burned patients. METHODS: Thirty-two severely burned patients were enrolled in the study with TBSA of 30.2 plus minus 10.9% and average burn index of 19.7. The everyday intake amounts of riboflavin, protein and calorie via different routes by each patient were recorded. Fluorescence method was employed to detect 4 hour urine load test in the 32 patients after oral taking of riboflavin on 3 similar 7 and 8 similar 14 postburn day (PBD). RESULTS: The averaged everyday intake amounts of protein and calorie by each patient in this study on 3 similar 7 and 8 similar 14 PBDs reached the requirement of Chinese burn patients' nutritional supplying formula. And protein administration occupied 17% similar 18% of total calorie. The averaged everyday intake of riboflavin by each patient was 3 similar 5 mg. But 4 hour urine load test exhibited that average urine discharging amount of riboflavin was all less than 400 microgram. CONCLUSION: There existed obvious systemic deficiency of riboflavin on 3 similar 14 PBD in severely burned patients. PMID- 11876876 TI - [The protective effects on postburn renal function by early enteral feeding in burned rats]. AB - OBJECTIVE: To study the effects of enteral feeding on postburn renal function in rats and its possible mechanism. METHODS: Wistar rats inflicted with 30% TBSA skin burn were adopted as the model. The effects of early enteral feeding (EEF) were dynamically observed on the changes of intra-gastric mucosal pH (pHi), portal endotoxin level, renal water content, plasma level of tumor necrosis factoralpha (TNFalpha), serum BUN and Cr contents and endogenous creatinine clearance rate. RESULTS: Compared with that in simple burn group, the pHi decrease could be inhibited by EEF, the portal endotoxin level, plasma TNFalpha level and renal water content were decreased by the action of EEF. Serum BUN, Cr contents were not affected by EEF. But the CCr increased by 4.67 times [from (16.43 +/- 2.9) to (3.52 +/- 0.79), P < 0.01] with EEF. CONCLUSION: EEF could obviously improve postburn renal function in severely burned rats, which might be related to the decrease of enteral endotoxin translocation and the postburn release of inflammatory mediators. PMID- 11876878 TI - [The dynamic changes of postburn complement activation and its effects on the secretory function of macrophages in severely burned mice]. AB - OBJECTIVE: To study the dynamic changes of postburn complement activation and its effects on the secretory function of macrophages in severely burned mice, so as to help explore the mechanism of postburn immunological disturbance. METHODS: The mice inflicted with 18% TBSA III degree scalding were employed as the model and randomly divided into normal control, decomplementizing control, scalding and decomplementizing scalding groups. The chemotactic activity of plasma, the production of nitride oxide (NO) and tumor necrosis factor alpha (TNFalpha) by peritoneal macrophages (MPhi) cultured by plasma from every groups were examined. RESULTS: The chemotactic activity of plasma complements increased quickly within 30 min. postburn, reached the summit level at 6 postburn hour (PBH) and declined since 12 PBH. Plasma from injured mice could induce peritoneal MPhis to produce more NO and TNFalpha. CONCLUSION: Complements were immediately activated postburn. The MPhis were activated by active complements. As a result, immunological active products increased significantly. PMID- 11876877 TI - [Analysis of the mental state of burn patients with postburn phobic neurosis]. AB - OBJECTIVE: To analyze the mental state of burn patients with postburn neurosis, so as to pave the way for solving the problem. METHODS: The clinical symptoms and the mental states of forty-three burn patients with phobic neurosis were analyzed in terms of age, sex, severity of burn injury and methods of burn management in accordance with phobic objects. RESULTS: (1) Simple phobia and circumstance phobia were mainly found in children, whereas sick phobia and social phobia were in adults. (2) Phobic neurosis happened more often in female, especially those above 12 years old, than in male patients. (3) The burn injury severity was positively correlated with the incidence of phobic neurosis. (4) There was higher incidence of phobic neurosis in patients treated with bandaging than those with exposure method. CONCLUSION: It was very important to close burn wound as soon as possible, to correct deformity, to improve dysfunction, to perform cosmesis and plastic surgery and to rebuild patients self-esteem in order to prevent or decrease the incidence of postburn phobic neurosis. PMID- 11876879 TI - [The effects of different patterns of mechanical ventilation on the respiratory mechanics and gas exchange in dogs severely injured by steam inhalation]. AB - OBJECTIVE: To study the effects of conventional mechanical ventilation (CMV), high-frequency jet ventilation (HFJV) and high-frequency bidirectional jet ventilation (HFTJV) on the respiratory mechanics and gas exchange in dogs inflicted severely by steam inhalation injury. METHODS: Nine mongrel dogs inflicted by steam inhalation to severe inhalation injury were served as the model. Indices of respiratory mechanics, hemodynamics and blood gas analysis were monitored respectively with the above three patterns of mechanical ventilation. RESULTS: (1) Total respiratory resistance (Rrs) and resistance of lungs (R(L)) were evidently higher with the ventilations than those in control (P < 0.01). And total respiratory compliance (Crs) and compliance of lungs (C(L)) were obviously lower with the ventilations than those in control (P < 0.05) but with no significant differences among the changes of Rrs, R(L), Crs and C(L). (2) Carbon dioxide (CO(2)) retention could be induced by CMV and HFJV. HFTJV was much more effective than HFJV in enhancing the expiration of CO(2) (P < 0.01). (3) There was no remarkable difference in functional residual capacity (FRC) and PaO2 between dogs with three patterns of ventilations. CONCLUSION: All these three patterns of ventilations could overcome the respiratory resistance and help respiration, but they couldn't restore the high airway resistance and low compliance of lungs already established postburn. The efficiency of CMV was similar to that by HFJV, but that of HFTJV was much better than that of HFJV. PMID- 11876880 TI - [An experimental study of myocardial injury by hydrofluoric acid in burned rabbits]. AB - OBJECTIVE: To investigate the myocardial injury by hydrofluoric acid in burned rabbits METHODS: New Zealand white rabbits were injured with 2% TBSA III degree skin burn on the back by hydrofluoric acid. The blood and tissue samples were taken from the rabbit after injury for the detection of myocardial and related enzymes and the pathomorphological examination. RESULTS: (1) Postburn myocardial enzymes increased significantly compared to those before burn (P < 0.01). (2) All the myocardial enzymes reached their peak levels within 48 postburn hours and were higher than those before burn (P < 0.01). (3) Among all the enzymes, CPK increased by 97.5% and those of LDH, AST and CK-MB by 81.9%, 76.1% and 60.3%, respectively. (4) Pathomorphological findings with LM were the granular degeneration and focal necrosis of myocardial fibers, interstitial hemorrhage of myocardium accompanied with neutrophil and lymphocyte infiltration. Ultrastructural changes included myofibril decrease or even disappearing, severe cellular and mitochondrial swelling, disappearing of mitochondrial crista, endothelial swelling, decrease of electronic density with the exhibiting of bulla and deformation of myocardial cellular nucleus. CONCLUSION: Myocardial function could be damaged after hydrofluoric acid burn. Therefore, special attention should be paid to the protection of cardiac function after hydrofluoric acid burn by strengthening the monitoring of myocardial enzymes and ECG, etc. PMID- 11876881 TI - [The effects of batroxobin on the healing of and microcirculatory blood flow volume in deep partial thickness burn wounds in rats]. AB - OBJECTIVE: To investigate the effects of batroxobin on the healing of and microcirculatory blood flow in deep partial thickness burn wound in rats. METHODS: Wistar rat inflicted by 4 cmx 4 cm deep partial thickness scalding on the back was taken as the model. Twenty male rats were randomly divided into scalding burn group and batroxobin treatment group. Cutaneous blood flow volume was measured before and at 0.5, 2, 4, 6, 12, 24 and 72 postburn hours (PBH). Scalding and residual burn wound areas were measured immediately and on the 14th and 18th postburn day (PBD). The rats were sacrificed on 30th PBD and the cutaneous samples were harvested for hair follicle counting and histological examination with LM. RESULTS: The cutaneous blood flow volume of rats in single burn group decreased progressively during 2-72 PBHs. But in batroxobin treating group, cutaneous blood flow volume improved evidently and the wound healing accelerated obviously with increased cutaneous appendages formation. CONCLUSION: Batroxobin might restore the blood circulation of stasis band of burn wound, so as to accelerate wound healing and to improve the quality of healed skin. PMID- 11876882 TI - [The effects of rapid fluid replacement on the hemodynamics of scalded shock dogs with delayed resuscitation]. AB - OBJECTIVE: To investigate the effects of rapid fluid replacement on the hemodynamics of scalded shock dogs with delayed resuscitation and to explore the protocol for the correction of postburn shock in case of delayed resuscitation. METHODS: Mongrel dogs inflicted with 40% TBSA III degree scalding were employed in the study. Twenty -- four dogs were randomly divided into 3 groups, i.e. thermal injury control group (C), delayed even fluid infusion group (E) and delayed rapid fluid infusion group (R). The femoral arterial pressure (FAP), pulmonary arterial pressure (PAP), pulmonary vascular resistance (PVP) and systemic vascular resistance (SVR) were monitored before and 2, 6, 8, 12, 24, 36 and 48 hours after injury. RESULTS: (1)All The dogs in C group died within 36 postburn hours (PBHs), while those in E and R groups survived more than 48 PBHs. (2) FAP and CO in all the dogs decreased obviously postburn while SVR increased evidently. After fluid infusion, FAP and CO bounced back while SVR decreased, especially in R group. (3) PAP, PAWP, CVP and PVR recovered rapidly but within normal range in R group. CONCLUSION: Rapid fluid resuscitation was beneficial and feasible for the correction of burn shock due to delayed resuscitation. PMID- 11876883 TI - [The effect of hyperoxic solution on acute myocardial hypoxia in scalded rabbits]. AB - OBJECTIVE: To investigate the effects of hyperoxic solution on acute myocardial hypoxia in scalded rabbits. METHODS: Rabbits inflicted with 30% TBSA III degree scald injury and/or acute myocardial ischemia were employed as the model. Animals were randomly divided into 4 groups, i.e. scalded rabbits treated with hyperoxic solution (A), scalded rabbits treated with normal saline (B), scalded rabbits without any treatment (C(b)) and normal healthy rabbits (C(h)). The cardiac function, blood rheology, blood gas analysis and the ratio of infarction to ischemic areas were dynamically monitored. RESULTS: (1) The cardiac function of the rabbits in B group was evidently worse than that in C(h) group (P < 0.01) and A group (P < 0.05). (2) The whole blood viscosity in A, C(b) and especially B groups was obviously increased compared with that in C(h) group (P < 0.05). While the viscosity in A group was much lower than that in B group (P < 0.01), it was similar to that in C(b) group (P > 0.05). (3) The PO(2) in A group was significantly higher than that in B and C (C(b), C(h)) groups (P < 0.01). (4) The ratio of infarction to ischemic areas in A group was remarkably lower than that in B group (P < 0.01). CONCLUSION: The hyperoxic solution may exert beneficial effects on acute myocardial ischemia in scalded rabbits. The mechanism might be related to the immediate correction of hypoxia in vital organs with simultaneous lowering of blood viscosity and the establishment of myocardial collateral circulation after the infusion of hyperoxic solution. PMID- 11876884 TI - [The postburn "shock heart"]. AB - OBJECTIVE: To investigate the pathogenesis of postburn "shock heart". METHODS: A combined serial studies in both animal experiments and prospective clinical investigation were carried out with the determination of indices reflecting myocardial injury, such as cardiac function, myocardial injury parameters, cytoplasmic and cytoskeletal structure injury, myocardial biomechanics and pathomorphological changes. The pathogenesis was also explored by the evaluation of myocardial nutritional blood flow (NBF), cardiac myocytic oxygen consumption, energy metabolism and cytokines released from cardiac myocytes. RESULTS: Cardiac function, myocardial biomechanics, NBF, the oxygen consumption, energy metabolism and the viscoelastic properties of cardiac myocytes decreased significantly postburn, and myocyte cytoplasm and cytoskeleton were damaged after burn. Furthermore, focal dissolution and fragmentation of myocardiac fibers were observed. In the meantime the plasma levels of CM -- LC, TnT and myocardial tissue cytokines increased markedly. CONCLUSION: These results suggest that organic damage of cardiac myocytes occurred during postburn shock stage. The principle pathogenesis might be related to ischemic -- reperfusion injury, inability of oxygen utility and energy metabolism in cardiac myocytes, as well as myocardial edema and uncontrolled inflammatory response. PMID- 11876885 TI - [The significance of the expression of the HSP70 and HSP90 in the intestinal mucosa in scalded rats during early postburn stage]. AB - OBJECTIVE: To investigate the changes in the expression, the tissue contents and the distribution of heat shock protein (HSP) 70 and HSP90 in the intestinal mucosa in scalded rats during early postburn stage and to elucidate the effects of heat shock response of intestinal mucosal cell on burn victim's systemic pathophysiology. METHODS: The scalded rats were taken as the model. ELISA, immunoblotting analysis and immunohistochemical methods were employed to analyze and study the expression, the tissue contents and the distribution and their functional status of HSP70 and HSP90 in the intestinal mucosa. RESULTS: (1) The free HSP70 content in the intestinal mucosa exhibited significantly transient decrease in the early postburn stage. (2) The total contents of HSP70 and HSP90 in the intestinal mucosa increased obviously postburn. (3) There exhibited remarkable heterogeneity of the molecular structure of HSP70 in the intestinal mucosa in the early postburn stage. CONCLUSION: The expression and the changes in the contents and the tissue distribution of the two HSPs in the intestinal mucosal cell might play important roles in the cellular stress reaction of the intestinal mucosa and even in the intestinal barrier functional mechanism. PMID- 11876886 TI - [The effects of escharectomy en masse during shock stage on the secretion of cytokines from macrophage in scalded rats]. AB - OBJECTIVE: To explore the effects of the plasma from rats undergoing escharectomy en masse during shock stage on the secretion of TNFalpha, IL-6 and IL-8 by macrophage. METHODS: Rats inflicted by 30% III degree scald were employed as the model. Sixty-three rats were randomly divided into escharectomy (E) and scald (C) groups. The plasma contents of TNFalpha, IL-6 and IL-8 in rats in two groups were determined. The plasma from rats in two groups was respectively co-cultured with rat peritoneal macrophages for 6 hours and the TNFalpha, IL-6 and IL-8 in the supernatants of the culture were assayed. RESULTS: Plasma contents of TNFalpha, IL-6 and IL-8 were evidently increased in both E and C groups. But the contents of these cytokines in E group decrease and were much lower than those in C group at 12 postburn hour. The sera from two groups of rats could all stimulate the secretion of TNFalpha, IL-6 and IL -- 8 from peritoneal macrophages, but the serum from C group possessed stronger stimulating ability than that from E group. CONCLUSION: Escharectomy en masse during shock stage could weaken the ability of burn sera to induce the secretion of TNFalpha, IL-6 and IL-8 by macrophages. It was therefore beneficial to the prevention of postburn development of systemic inflammatory response syndrome. PMID- 11876887 TI - [The clinical and experimental study on postburn metabolic characteristics of zinc and its influence on copper, iron and calcium]. AB - OBJECTIVE: To observe postburn metabolic characteristics of zinc and its influence on copper, iron, calcium. METHODS: (1) To analyze the levels of Zn, Cu, Fe, Ca in the serum and urine of burn patient on different postburn days. (2) To replicate a rat model of zinc deficiency and to observe the symptoms and the effects of zinc supplementation in different dosage after scalding in rats with zinc deficiency. (3) To assay the levels of Zn, Cu, Fe, Ca in rat wound tissues following supplementation of zinc in diet and to wounds in rats after scald. RESULTS: (1) The levels of serum Zn and Cu decreased markedly on 1 similar 3 days in major burn patients, whereas the urine Zn and Cu increased obviously, reaching the peak value on the 14th day. The serum and urine Fe increased rapidly on PBD2 and then descended steeply. The serum and urine Ca kept lower levels all along. (2) In zinc deficient rats growth retardation appeared first and other symptoms emerged subsequently. Serum Zn decreased and serum Cu, Fe, Ca increased. (3) The levels of Zn, Cu, Fe, Ca in wound tissue of scalded rats showed a continuous postburn increase. The Ca contents in wound tissue increased about 50 -- fold. Zn content in wound was more obviously increased than Cu, Fe and Ca when Zn was supplemented through wound compared with that with diet supplementation. CONCLUSION: Serum Zn level was lowered after scald and it could be elevated after supplementing zinc by diet or through wound, Changes in serum Cu, Fe, Ca occurred corresponding to the change in serum Zn. PMID- 11876888 TI - [The effect of Shenmai injection on the expression of TNFalpha mRNA of the macrophages in scalded mice]. AB - OBJECTIVE: To explore the effect of Shenmai injection on the expression of TNFalpha mRNA of peritoneal macrophages in scalded mice. METHODS: BALB/c mice inflicted with 11% TBSA III degree scalding injury were employed as the model. Shenmai injection was composed of 100 mg of red ginseng and 100 mg of lily turf root in 1 ml injection solution and was administered to mice via peritoneal injection in a dose of 2.5 ml per kilogram of mice body weight per day for 5 days. The expression of TNFalpha mRNA was determined by semi-quantitative RT-PCR. RESULTS: The expression of macrophage TNFalpha mRNA was significantly increased in the scalded mice, while the expression was obviously lowered after the administration of Shenmai injection (P < 0.01) to scalded mice and the survival rate of the scalded mice was thus markedly increased (P < 0.05). CONCLUSION: The beneficial effect of Shenmai injection on the scalded mice might be related to its inhibitory effect on the hyperexpression of macrophage TNFalpha mRNA. Shenmai injection could lower down the expression of TNFalpha in scalded mice and raise the survival rate of scalded mice. PMID- 11876889 TI - [Local expression of interleukin-12 mRNA in grafted skin in rats]. AB - OBJECTIVE: To investigate the immunological modulating effects of IL-12 in local tissue of grafted skin. METHODS: The rats grafted with intermingled (allo-auto), allo and auto skins were employed as the model. The expression of IL-12 (P35, P40) mRNA in local tissue cells of grafted skin was determined at different time points. RESULTS: There was evident difference (P < 0.05) of the expression of IL 12 (P35) mRNA between the rats in allo grafting and auto grafting groups. There existed obvious differences of the expression between all the values in every time points in rats of intermingled grafting group except for that in 14 PBD (P < 0.05). There were significant differences of the expressions between the rats in intermingled and auto grafting groups at 4, 7, 14 and 21PBDs (P < 0.05). There were marked differences of the expression of IL-12 (P40) mRNA between the rats in allo and auto grafting groups before 21 PBD (P < 0.05). When compared with that in the rats of intermingled grafting group, the expression in allo grafting group was only different at 4 PBD (P < 0.05). There were obvious differences of the expression (P < 0.05) between the rats in intermingled and auto grafting groups on 7, 14 and 21 PBDs. During the rejection process of allo and intermingled grafting, the rule of the expression of IL-12 mRNA was in accordance with clinical observation and pathomorphological dynamic detecting results. CONCLUSION: IL-12 actively played a part in inflammatory reaction and played an important role in the conversion of T helper cell to type I T helper cell (TH1) and in mediating cellular immune process. There might be immunosuppressive effects mediated by type II T helper cell (TH2) cytokines in the intermingled skin grafts. PMID- 11876890 TI - [The effects of the receptor antagonist of glutamic acid on the brain tissue of severely scalded rats]. AB - OBJECTIVE: To explore the influence of glutamic acid on burned rat brain tissue and the effects of the receptor antagonist of glutamic acid. METHODS: Rats inflicted with 30% TBSA III degree scalding injury were taken as the model. The water content, K(+), Na(+), Ca(2+), Mg(2+), nitric oxide metabolites (NO(2)( )/NO(3)(minus sign))concentrations of scalded rat brain were determined at 2, 6, 12 and 24 postburn hours (PBH). Morphological and histochemical studies of the brain tissue were carried out with LM and EM. RESULTS: The water content, Na(+), Ca(2+)and NO(2)(minus sign)/NO(3)(minus sign) concentrations of the brain tissue after scalding injury were higher than those in control group. But intracerebral ATP enzyme decreased postburn. It was found by EM examination that there existed swelling of capillary endothelium, nerve cells and some intracytoplasmic mitochondria. There appeared pinosome on capillary wall. After the administration of receptor antagonist of glutamic acid, D-2-amino-group-7-phosphoenanthate (D AP7), the brain water content, Na(+), Ca(2+)and NO(2)(-)/NO(3)(-) concentrations in brain tissue, and the capillary wall pinosome decreased, and the swelling of nerve cells ameliorated. CONCLUSION: There might be postburn increased cerebral microvascular permeability, ischemic injury of cerebral tissue cells and the development of brain edema. The postburn alterations in the morphology and metabolism of brain tissue were correlated with the over secretion of glutamic acid. And the excitatory toxic effects of glutamic acid were mediated by its receptor. The receptor antagonist of glutamic acid D-AP7 might ameliorate postburn cerebral injury. PMID- 11876891 TI - [The significance and characteristics of the gene expressions of c-fos and c-jun in hypertrophic scar and chronic ulcer tissues]. AB - OBJECTIVE: To explore the characteristics and regularity of the expression of c fos and c-jun genes in hypertrophic scar and chronic ulcer tissues and their relationship to different tissue restoration. METHODS: Tissue samples of hypertrophic scars were harvested from 16 cases of patients after burns and chronic cutaneous ulcers during operation together with the normal skin (n = 5) of the same patients as control. ABC method was employed to detect the gene expression of both c- fos and c-jun in above samples. RESULTS: The positive expression of c-fos and c-jun gene was found in epithelial basal cells and some subcutaneous fibroblasts in normal skin. But the expression of c-jun was weaker than that of c-fos in above tissues. A strong positive expression of c-fos and c jun genes was found mainly in fibroblasts in the hypertrophic scar. But the combined expression of both tumor genes was most often found in the capillary endothelium, some inflammatory cells and the cytoplasm of fibroblasts in ulcer tissue. CONCLUSION: The results indicated that the amount and location of the expression of c-fos and c-jun genes were different in the three kinds of tissues, suggesting that these two tumor genes may play important roles in wound healing modulation. PMID- 11876892 TI - [Analysis of the long-term results of late hand function reconstruction in patients with severe electrical injury of the wrists]. AB - OBJECTIVE: To evaluate and analyze the long-term results of all the operation mordalities practised at present of late hand function reconstruction in patients with severe electrical injury of the wrists. METHODS: Forty-seven cases with late electrical injury of the wrists were followed up and comprehensively analyzed in terms of the long-term results of morphological reconstruction and functional restoration. RESULTS: In addition to cutaneous tissue repair and functional training of the joints of the wrists, the flexor digitorum reconstruction with auto free tendon grafting gave a poor result (only 33.4% of the cases with function above medium level). In one case, the wrist function was rebuilt by compound plantaris tendon free flap with leg deep facia and posterior tibial artery, resulting in the restoration of thumb flexor function, with shorter treating time and less postoperative adhesion. Median and ulnar nerves were repaired with free auto nerve grafting with no obvious effects. Two cases were repaired by compound tissue grafting of sural nerve and deep facia carried by small saphenous veins with uncertain result. In another 3 cases, with the aid of retrograde evoked potential (REP), a proximal anastomosis of nerves was done with fairly good result. CONCLUSION: Insufficient blood supply and scar formation were the major causes of poor results of the tendon and nerve grafting. It was recommended that the graft should carry sufficient blood supply to improve the blood supply of the recipient bed in order to improve the therapeutic effects. The selection of proximal nerve segment should include the functional examination. PMID- 11876893 TI - [Clinical application of irradiated drug-containing porcine-cornea to patients with ocular burns]. AB - OBJECTIVE: To explore a new method for the management of patients with ocular burns. METHODS: Fifty-five cases of patients with ocular burns (in 88 eyes) were randomly divided into treatment and control groups. Thirty cases in treatment group with 49 eyes were transplanted with irradiated drug-containing (ofloxacin, acetyl cysteine and reduced glutathione) porcine-cornea. 25 cases in control group with 39 eyes were treated with routine program. RESULTS: Thirty-two eyes were rescued in treatment group with the cure rate of 65.3%. But only 17 eyes were saved in control group with the cure rate of 43.59%, indicating significant difference of the cure rate between the two groups (P < 0.05). CONCLUSION: Irradiated drug-containing porcine-cornea might well be an ideal therapeutic material for the management of patients with ocular burns. PMID- 11876894 TI - [Morphological changes in canine heart after electrical injury]. AB - OBJECTIVE: To observe the histomorphological changes in canine heart after electrical injury. METHODS: Light and electronic microscopes (LM, EM) were employed to observe the histomorphological changes in canine heart after electrical injury. RESULTS: There were swelling and deformation of pacemaker cells (p cells) and mitochondrial swelling and vacuolization in cardiac conduct system. The cardiac muscle cells in the atrial and the ventricular walls exhibited obvious swelling and deformation, blurred cross striation, intracytoplasmic structural destruction of myofibrils, messy microfilaments, mitochondrial swelling, vacuolization and even dissolution. Furthermore, there was also breaking and expanding of the intercalated disk between adjacent cardiac muscle cells. The gap junctions at sites of intercalated disk were disrupted. CONCLUSION: The pathomorphological characteristics of cardiac histological changes after electrical injury were similar to those of acute myocardial infarction. PMID- 11876895 TI - [Study on the abnormal gene expression of inducible nitric oxide synthase (iNOS) in rabbit myocardium after high voltage electrical injury]. AB - OBJECTIVE: To elucidate the effects of high voltage electrical injury on the mRNA expression of iNOS And endothelin-1 (ET-1) in rabbit myocardium. METHODS: Rabbits inflicted by high voltage electrical injury were employed as the model. Reverse transcription PCR (RT-PCR) and serum enzyme measurement were used in the detection of the expression levels of iNOS mRNA and ET-1 mRNA of rabbit myocardial tissue and plasma creatine kinase (CK) content. RESULTS: The expression of iNOS mRNA in rabbit myocardial tissue after high voltage electrical injury increased at 6 to 8 postburn hours (PBH) and decreased thereafter. The plasma contents of CK and ET-1 mRNA reached their peak values at 2 PBH. CONCLUSION: The increase of iNOS mRNA might protect myocardial tissue from electrical injury. PMID- 11876896 TI - [Establishment of a rabbit model of non-thermal high voltage electrical injury]. AB - OBJECTIVE: To establish a rabbit model of non-thermal high voltage electrical injury, which was accompanied by progressive tissue necrosis for the further study of electrical injury. METHODS: Seventy-five New Zealand rabbits were employed, in which 45 were used for the selection of the size of electrode plate, the inflicting time, the intervals and the injury degree. Five groups of rabbits were used for the study of the model. The study was carried out by means of clinical anatomical exploration, categorization by the Index of Deep Burn Injury (IDBI) and (99)Tc(m)-MDP isotope scanning and gamma photography. RESULTS: The optimal injury indices selected were as follows: the electric field strength was 17 000 volts/m, the mean current intensity was 554 mA, and the average current density was 137 mA/cm(2) for small electrode and 21 mA/cm(2) for big one, and average increase of tissue temperature was 1.73 degrees C during injury process. This excluded the possibility of thermal injury. Five models were created, i.e. mild, moderate, severe, extra severe and destructive ones. There was no obvious cutaneous necrosis. Nevertheless, there was loss of injured extremities on 5th, 7th and 12th post-injury days in the severe, extra severe and destructive groups. CONCLUSION: Non-thermal factor was the major cause of electric injury in the model with typical clinical features. PMID- 11876897 TI - [Histochemical study on the nitric oxide synthase in the intestinal myenteric plexus of burned rats]. AB - OBJECTIVE: To elucidate the postburn change in the activity and the distribution of nitric oxide synthase (NOS) in rat intestinal myenteric plexus. METHODS: The activity and distribution of NOS were quantitatively and locationally studied by histochemical staining with NADPH-diaphorase (NDP), a marker for NOS, and whole mount preparation technique. RESULTS: NOS was widely distributed in whole intestine wall, especially in myenteric plexus. The majority of NOS positive neurons displayed either round or oval shape. There were some varicosities string of bead-like structure in the nerve fibers and which were accompanied by blood vessels and muscular fibers. There was no significant postburn change in the density of NOS positive neurons in myenteric plexus (P < 0.05). Nevertheless the NOS activity of neurons decreased obviously (P < 0.05 similar 0.01). Meanwhile, it was found after injury that there were indistinction of the structure of NOS positive neuron, more breaking of nerve fibers, leading to the lack of integral web of nerve fibers and obvious decrease of NOS positive varicosities in nerve fibers. CONCLUSION: After burn injury, the NOS positive neural structure in myenteric plexus of rat small intestine was damaged with the decrease in NOS activity. Simultaneously, there was impairment of the releasing route of nitric oxide (NO). All these changes might be closely related to intestinal structural damage and dysfunction after burn injury. PMID- 11876898 TI - [The influence of hTGFbeta(1) gene transfection on the proliferation of NIH 3T3 fibroblast]. AB - OBJECTIVE: To investigate the influence of hTGFbeta(1) ene transfection on the proliferation of in vitro cultured NIH 3T3 fibroblast. METHODS: Lipofictin mediated co-transfected method was employed to transfect hTGFbeta(1) plasmid DNA into NIH 3T3 fibroblasts of the rats. Cell growth counting method, MTT, flow cytometry (FCM) and soft agar clone forming method were adopted to examine the biology of the transfected cells. RESULTS: (1) Fibroblast growth slowed down after hTGFbeta(1) transfection, especially on 4th to 6th days after the transfection, which was accompanied by decreased synthesis of DNA as indicated by the increase of G(1) phase percentage (from 39.9% to 66.2% P < 0.05) and the decrease of S phase percentage (from 40.2% to 26.8%, P < 0.05); (2) There was no evident change of all the phases in a cell cycle in the blank load transfection group and in control group. (3) There was a good correlation between MTT and cell counting method (co-efficient 0.992) (4) The formation rate of the soft agar clone of fibroblast decreased from 1.18% to 0.55% (P < 0.05) after hTGFbeta(1) transfection. CONCLUSION: hTGFbeta(1) gene transfection could inhibit fibroblast proliferation, which might be related to the interference of TGFbeta(1) with the DNA synthesis of fibroblasts. PMID- 11876899 TI - [Study on the pharmacokinetics of Imipenem cilastatin in burn patients during the acute phase]. AB - OBJECTIVE: To investigate the pharmacokinetic characteristics of Imipenem/cilastatin in burn patients during the acute phase. METHODS: Imipenem concentrations in the plasma, blister and, interstitial fluids and urine of 6 burn patients (P group) were determined during the acute phase by high performance liquid chromatography after the first initial dose. Pharmacokinetic parameters were thus produced and statistically analyzed by program package STAT 5. Ten healthy volunteers served as control group (C group). RESULTS: Compared to those in control group, pharmacokinetic parameters of Imipenem exhibited evident difference, such as prolonged half-life [(1.56 plus minus 0.55) h vs (1.06 plus minus 0.24) h] and enlarged distributing volume (16.16 plus minus 4.26) vs (13.96 plus minus 7.10). Imipenem could be detected in the blister and interstitial fluids of burn patients 1 hr after the initial dose. Renal clearance of Imipenem was positively correlated to creatinine clearance (r = 0.5834). CONCLUSION: The intervals between doses should better be prolonged when loaded-dosage of Imipenem was administered in burn patients during the acute phase. Imipenem could penetrate II and III degree burn wound when given in the acute phase. Attention should be paid to renal function when Imipenem was given. PMID- 11876900 TI - [The postburn change in splenic T lymphocyte transmembrane signal transduction and its relationship with the secretion of IL-2 and IL-10 in severely scalded mice]. AB - OBJECTIVE: To explore the postburn functional change in splenic T lymphocytes and the secretory rules of IL-2 and IL-10 in severely scalded mice, and to look for the explanation of the change by means of T lymphocyte transmembrane signal transdunction study. METHODS: The changes in the antigen receptors on T cells (TCRalpha/beta), assisting stimulating molecule (CD28) and the activities of GTPase, protein tyrosine kinase (PTK) and protein kinase C (PKC) that participated in the transmembrane signal transduction were detected, so as to observe the proliferation and transformation function of T cells at different post-burn periods and the secretion of IL-2 and IL-10. Furthermore, the changes in every signal transduction molecules were analyzed with the consideration of their relationship with the change in T cell function activities. RESULTS: The postburn positive rates of the expressions of TCRalpha/beta and CD28 on T cell membrane decreased. The postburn GTPase activity and membrane PTK activity were all suppressed but recovered at 168 postburn hours. Membrane PKC activity exhibited a dual-phase change (increase following decrease), which was closely related to IL-10 level. CONCLUSION: The postburn changes in TCRalpha/beta, CD28 on T cell membrane molecules and transmembrane signal transduction enzymes were important factors contributing to the decrease in IL-2 secretion, suppressed T cell function and the dual-directional changes in IL-10 secretion. PMID- 11876901 TI - [A comparative study on wound healing treated by different doses of bovine basic fibroblast growth factor (bFGF)]. AB - OBJECTIVE: To explore the optimal dosage and method of bFGF application on surgical wound for accelerating its healing. METHODS: Laceration wounds on the extremities of 73 patients were served as the target. The wounds were applied with three different doses of bFGF (75 Au/cm(2), 150 Au/cm(2), 300 Au/cm(2)) and with three application methods, i.e. once a day, three times a day and once a day with moistening. Normal saline with 0.5% metronidazol was employed as control agents. The wound healing time in every groups was recorded in days and compared with each other. RESULTS: When compared to each other, the wound healing time (day) was much shorter (P < 0.05) with bFGF application in dose of 75 similar 300 Au/cm(2). There were also obvious differences in wound healing time among groups treated with the same dose but with different methods (P < 0.01). In addition, three times a day and once a day with moistening of bFGF application could further shorten the wound healing time. CONCLUSION: Compared to the dosage of the agent, the frequency and method of bFGF application seemed more important for its optimal effects. In view of the convenience, feasibility and economy, once a day with moistening was recommended for clinical application. PMID- 11876902 TI - [The effects of DiBaiRen decoction on the proliferation of human keratinocytes and its in vitro anti--oxidation role]. AB - OBJECTIVE: To investigate the effects of DiBaiRen decoction on the proliferation of human keratinocytes and its in vitro anti -- oxidation role. METHODS: Human keratinocytes were cultured in vitro in the media containing gradient diluted DiBaiRen decoction. The cell proliferation rates at different culture times were recorded. Cellular oxidation injury inflicted by superoxide anion and hydroxyl anion was produced by adding hypoxanthine -- xanthine oxidase and hydrogen peroxide to the culture media. MTT colorimetry was employed to determine cellular activity. The antagonistic effects of DiBaiRen decoction on cellular oxidation injury were analyzed. RESULTS: In the case that DiBaiRen decoction was added to the culture in concentrations of 30 ml/L to 100 ml/L, there exhibited obvious accelerated proliferation of the keratinocytes, with evident shortening of the cell fusion time. In addition, the oxidation injury of cultured keratinocytes caused by hypoxanthine -- xanthine oxidase and hydrogen peroxide was ameliorated or eliminated by the addition of DiBaiRen decoction, and the effects were dose -- dependent. CONCLUSION: DiBaiRen decoction might possess anti -- oxidation effects and could promote the in vitro proliferation of cultured human keratinocytes, and it might be beneficial in wound protection and epithelization. PMID- 11876903 TI - [The management of bony burn with skin flap transplant with preservation of sequestrum]. AB - OBJECTIVE: To study the management of bony burn, so as to improve the quality of bony burn treatment, to decrease the complications and to shorten the course. METHODS: Retrospective review of 36 cases with bony burn cured since 1992 was done in terms of treatment strategy before skin flap transplant to cover the burn wound, i e sequestrum preserved or partial removal. RESULTS: There exhibited no complications, such as infection, osteomyelitis and pathologic bone fracture in all these cases. CONCLUSION: The treatment strategy described here could be advocated in view of its reliable coverage of the wound, shortening of the bony burn course and reduction of complication. PMID- 11876904 TI - [Retrospective clinical analysis of the emergent comprehensive repair of electrical injury]. AB - OBJECTIVE: To summarize the experience with clinical emergent comprehensive repair of electrical injury. METHODS: One hundred fifty five cases of electrical injury during past 14 years were analyzed in terms of the emergent repair methods on the basis of medical records and photographs. RESULTS: In this group of patients the average interval between injury and hospitalization was 14.9 +/- 21.5 hours, And 108 cases (70%) were admitted to hospital within 2 hours post injury. The average time between admission and first operation was 3.74 +/- 9.89 hours, and 70% of them was within 1 hour after admission. Four hundred and fifty nine wounds were debrided and 296 strips of tendons, 19 strips of blood vessels, 196 pieces of bone or joints and 16 strips of nerves, which were exposed, devitalized or necrotic, were saved. The grafting repair of tendons, bones, nerves and blood vessels was done in 24 case times. Three hundred and ninty eight tissue flaps were employed in 459 wounds. CLS (NS + Lidocaine + amphicol) solution was employed to irrigate underneath the flaps for 24 similar 96 hours or longer. Composite dan -- shen root (scatellaira indica) solution was given to all the patients by injection. As a result, the excellent rate of function and contour was 84.8%. The unsatisfactory and amputation rate was 9.6%. The rate of extremity and/or finer amputation was 1.8%. The rate of total limb loss including those lost instantly after injury was 9%. CONCLUSION: Emergent comprehensive management of electrical injury was a feasible and effective method for decreasing extremity loss. PMID- 11876905 TI - [The hEGF expression of gene--transferred human epithelium after its grafting on porcine surgical wound]. AB - OBJECTIVE: To investigate the hEGF expression of gene -- transferred human epithelium after its grafting on porcine surgical wound. METHODS: Gene -- transferred and normal epithelia were implanted on the biological silicone membrane and then grafted on porcine surgical wound. The tissue samples were harvested from the wound and detected for the hEGF expression with immunohistochemical method on 1, 3, 5, 8 and 12 post -- grafting days. RESULTS: There existed positive hEGF expression in the wound grafted with gene -- transferred epithelia and negative hEGF expression in the wound grafted with normal epithelium. CONCLUSION: Gene -- transferred epithelium could express hEGF on the grafted wound. PMID- 11876906 TI - [A study on the detection of apoptosis of hypertrophic scar and its related modulating factors]. AB - OBJECTIVE: To investigate the relationships between the development of hypertrophic scar (HS) and apoptosis of fibroblast and endothelial cell, and to elucidate the influence of related modulating factors. METHODS: TUNEL and immunohistochemistry staining techniques were employed to examine apoptosis of fibroblast and endothelial cell. The expression of ICE and Bcl-2 in hypertrophic scar of 61 cases of postburn patients undergoing plastic surgery with HS and in 20 other operative patients with non-hypertrophic scars (NHS) was also determined. RESULTS: The positive apoptotic cell indices of fibroblast and vascular endothelial cells during the proliferative stage of HS were 6.60 plus minus 4.43 and 8.90 plus minus 6.01 respectively, while those during the mature stage were 25.60 plus minus 5.70 and and 26.6 plus minus 6.02, respectively. There existed evident differences in positive apoptotic cell indices between the same kind of cells (fibroblasts or vascular endothelial cells) during the two stage (P < 0.01). The ICE positive expression rate in proliferative stage was much lower than that in mature stage (P < 0.01), but the Bcl-2 positive expression rate in proliferative stage was obviously higher than that in mature stage (P < 0.01). CONCLUSION: HS development might be related to the decrement of apoptotic fibroblasts and endothelial cells. ICE and Bcl-2 were possibly involved in the modulation of fibroblast and endothelial cell apoptosis in HS. PMID- 11876907 TI - [An experimental study on the effects of NGF on the wound healing of deep partial thickness burn in pigs]. AB - OBJECTIVE: To investigate the effects of nerve growth factor (NGF) on wound healing of deep partial thickness burn in pigs. METHODS: Six pigs of about 20 kg each were employed as the model. Twenty -- four deep partial thickness burn wounds (2.5 cm in diameter per wound) were made on the back of each pig by temperature and pressure controller. Six wounds were set to be one group. The wounds were randomly divided into 4 groups, i.e. control group (applied with normal saline, C), three treatment groups with NGF topically applied in doses of 1, 2.5 and 5 microgram/ml, respectively. Histological examination, determination of hydroxyproline, analysis of cellular DNA cycle and wound healing time in every groups were carried out on 3, 5 and 9 postburn days (PBD). RESULTS: The proliferation rate of epithelia was much more active in treatment groups than in control group. Epithelization of the wound in treatment groups was earlier than of that in control group. The hydroxyproline content in treatment groups exhibited an increment after a prior decrement, especially on 5 PBD when the content was much lower in the treatment groups. The analysis of cellular DNA cycle indicated that cell number of S phase in treatment groups was obviously more than that in the control group. Furthermore, the wound healing time in treatment groups was much shorter than that in control group. CONCLUSION: Local application of NGF might promote the wound healing of deep partial thickness burn. PMID- 11876908 TI - [The role of p38 MAPK in LPS induced ICAM-1 expression on endothelial cell]. AB - OBJECTIVE: To study the role of p38 mitogen-activated protein kinase (MAPK) in lipopolysaccharide (LPS) induced expression of intercellular adhesion molecule-1 (ICAM-1) on human umbilical vein endothelial cell (HUVEC). METHODS: HUVEC was harvested and cultured before being divided into two groups, i,e. stimulating group (S) and priming group (P). In S group, the cultured endothelial cell was stimulated by LPS. In P group, endothelial cell was pre-treated with SB 203580 2 hours before LPS stimulation. The expressions of ICAM-1 mRNA and protein in HUVEC of two groups and their dose-effect relationship with stimulator and inhibitor were observed. Furthermore the p38 mRNA activity of HUVEC was detected. RESULTS: After LPS stimulation, ICAM-1 molecule on the surface of endothelium increased significantly at 8 to 36 hours. Cytoplasmic mRNA increased obviously at 2 hours. The activity of p38 MAPK increased at 15 min and reached peak value at 30 to 60 min after that HUVEC was stimulated by LPS. CONCLUSION: Inhibitor SB 203580 of p38 might significantly inhibit the inducing effect of LPS. LPS could regulate the expression of ICAM-1 gene and protein in HUVEC by activating p38 MAPK signal transduction passage. PMID- 11876910 TI - [The effects of ecdysterone on the apoptosis and necrosis of cultured HUVEC induced by endotoxin]. AB - OBJECTIVE: To study the protective effects of ecdysterone (EDS) on the apoptosis and necrosis of cultured HUVEC induced by endotoxin. METHODS: HUVEC was cultured in vitro. Apoptosis and necrosis of HUVEC were induced by LPS, and EDS was added in test specimen. Apoptosis was determined by means of flow cytometry and fluorescent microscopy. RESULTS: There was relatively less number of apoptosis and necrosis of in vitro cultured normal HUVEC than that after LPS stimulation, whereas apoptosis dominated over necrosis. After EDS was added to the culture of HUVEC, cell apoptosis and necrosis decreased simultaneously. CONCLUSION: EDS might exert protective effects on cultured HUVEC against apoptosis and necrosis induced by LPS. PMID- 11876909 TI - [A study on the mechanism of the biological roles of danshensu on fibroblast]. AB - OBJECTIVE: To explore the effects of danshensu on fibroblast apoptosis and the expression of procollagen gene. METHODS: Danshensu was added to in vitro culture of human cutaneous fibroblasts, and the nuclear proteins and total RNA were harvested from the cells. Electrophoretic mobility shift assay (EMSA) was adopted to determine the combining activity of nuclear transcription factors NF-kB and NF 1. Cell apoptosis was analyzed by DNA ladder fragmentation. The procollagen gene expression was analyzed by RT-PCR. RESULTS: Eight hours after the action of danshensu on the culture fibroblasts, the NF-kB combining activity was almost inhibited completely and the NF-1 combining activity decreased by about 50%. In addition, ladder-like DNA fragments were revealed clearly by sepharose electrophoresis. The mRNA levels of type I procollagen alpha1 and alpha2 decreased by 56% and 59%, respectively. CONCLUSION: Danshensu could inhibit the nuclear transcription factor NF-kB activity of fibroblast and induce the occurrence of its apoptosis. Furthermore, danshensu could also inhibit the nuclear transcription factor NF-1 activity of fibroblast and modulate the synthesis and secretion of collagen. PMID- 11876911 TI - [The significance of the postburn expression of proto -- oncogenes c -- fos and c -- myc mRNA and proteins in rat myocardial cells]. AB - OBJECTIVE: To investigate the postburn expression of proto -- oncogenes c -- fos and c -- myc mRNA and proteins in rat myocardial cells following severe burn. METHODS: Wistar rats inflicted with 30% TBSA III degree burn on the back were employed as the model and randomly divided into four groups: burn (B), resuscitation (R), verapamil (V) and control (C) groups. The expressions of c -- fos and c -- myc mRNA were examined by in situ hybridization and that of the proteins by immunohistochemistry. RESULTS: The expressions of the proteins and mRNA of c -- fos and c -- myc in every experimental groups increased to peak values and then decreased along with time lapsing. And the expressions of c -- fos and c -- myc were weakest in B group, while that in V group was stronger than that in R group. CONCLUSION: The expression of postburn immediate -- early genes c -- fos and c -- myc could be induced in the myocardial cells in all the groups by severe burns. And the expression features were different from the gene expression in patients with myocardial infarction and with cardiac over -- load. This implied that there were different signal transduction and modulation mechanism among these pathological conditions. PMID- 11876912 TI - [The protective effects of Zn-metallothionein on severely scalded rats inflicted by oxygen free radicals]. AB - OBJECTIVE: To investigate the protective effects of metallothionein (MT) on severely scalded rats inflicted by oxygen free radicals after delayed resuscitation. METHODS: Wistar rats inflicted by 30% TBSA III degree scalding injury were taken as the model. Fifty-Six rats were randomly divided into four groups, i.e. normal control, delayed resuscitation, MT treated and VitC treated groups. The changes in superoxide dismutase (SOD) and malondialdehyde (MDA) contents in plasma and subeschar edematous fluid were determined at 24 and 48 postburn hours with traditional indirect detecting method and electron spin resonance (ESR). Pathomorphological examination was simultaneously carried out for cardiac, hepatic, renal and intestinal samples. Plasma biochemical indices were determined at the same time. RESULTS: In delayed resuscitation group, SOD content decreased and the MDA contents increased in plasma and subeschar fluid obviously, with remarkable changes in the pathomorphology of all the internal organs and the blood biochemical indices. But in MT treating group, SOD content increased (P < 0.05) with decreased content of MDA (P < 0.05) and the pathomorphology and blood biochemical indices improved significantly when compared with those in delayed resuscitation group and VitC treatment group. CONCLUSION: MT treatment might be beneficial in the management of severely scalding rats infliced by oxygen free radicals. PMID- 11876914 TI - [An experimental study on the release of endotoxin from gram negative bacteria induced by antibiotics]. AB - OBJECTIVE: To explore the characteristics and possible mechanism of LPS released from Gram negative bacteria induced by antibiotics, so as to improve clinical management of endotoxemia and sepsis. METHODS: Cultures containing PA103 subtype of Pseudomonas aeruginosa (PA) and E coli 25922 subtype of E coli were treated with four kinds of antibiotics as Imipenam (IMP), ceftazidime (CTZ), amikacin (AMN) and pefloxacine (PFX) in four concentrations of 0.5, 1, 5 and 10 MIC for 8 hours. The changes in the bacterial quantity and morphology and the supernatant levels of free LPS of the culture media were observed at different time points. RESULTS: All the four kinds of antibiotics could kill the tested bacteria in similar degree, but lead to the different types of morphological changes of the bacteria. In detail, IMP could convert the bacteria into spherical shape, while CTZ and PFX made the bacteria to filamentous shape. But AMN could induce lysis of bacterial thallus. Under same condition, the ability of different kinds and concentrations of antibiotics to induce LPS release ranked as CTZ > PFX > IMP > AMN, 0.5MIC > 1MIC > 5MIC > 10MIC. Along with the prolongation of the action time, the LPS release increased. Furthermore, PA103 released less endotoxin than E. coli after the action of antibiotics. CONCLUSION: All of the four antibiotics, i,e, IMP, CTZ, AMN and PFX could induce PA103 and E coli 25922 to release different levels of LPS, which was related to bacterial morphological changes. The LPS release from the bacteria was correlated to the antibiotics applied, concentrations, action time and the bacterial features. Antibiotics with less ability of inducing LPS release were recommended for clinical management of the sepsis and/or septic shock caused by Gram negative bacteria. PMID- 11876913 TI - [Modulation of the activity of human alpha1 (I) procollagen gene promoter by basic fibroblast growth factor (bFGF)]. AB - OBJECTIVE: To investigate the modulating effect of bFGF on the proliferation of human cutaneous fibroblasts and on the activity of human alpha1 (I) procollagen gene promoter. METHODS: Human cutaneous fibroblasts were cultured and subcultured by tissue block culture technique. The influence of different concentrations of bFGF on the proliferation of fibroblasts was determined by ELISA method with BrdU incorporated into fibroblast DNA. Three plasmids containing various lengths of 5prime prime or minute flank sequences of human alpha1 (I) procollagen gene and CAT as reporter gene were constructed and transfected into the fibroblasts by FuGENE transfection reagent. The quantitative expression of the fibroblast CAT was determined by ELISA after treatment with bFGF. RESULTS: After 24 hours of treatment of the fibroblasts by serial concentrations (0.25 ng/ml similar 64.00 ng/ml) of bFGF in DMEM containing 2% (v/v) or 10% (v/v) FCS, the BrdU incorporation into DNA was determined. The proliferating rate of the fibroblasts differed significantly from each other in all the groups (P < 0.05). After the fibroblasts were transfected with the three plasmids and treated thereafter by 4 ng/ml and 16 ng/ml of bFGF for 24 hours, the relative CAT expression values were determined. It indicated that the expression value was evidently different between bFGF processing and control groups (P < 0.05). CONCLUSION: bFGF might inhibit the proliferation of human cutaneous fibroblasts and exert negative regulating effect on the human alpha1 (I) procollagen gene promoter sequence in dose -- dependent pattern. PMID- 11876915 TI - [Micro -- ecological investigation of burn wound infection]. AB - OBJECTIVE: To analyze the epidemic tendency of the bacteria causing burn wound infection, and to explore the influencing factors. METHODS: Bacterial ecology on burn wound was investigated in 210 cases of burn patients admitted to our hospital from January 1999 to December 2000, and the results were compared to those done in the 1970s, 1980s and 1990s. RESULTS: Since 1999, G(+)cocci increased obviously (52.17%) when compared with those of the past three decades (P < 0.01), while G(minus sign) bacilli decreased significantly (46.48%, P < 0.01). The ratio of Pseudomonas aeruginosa in recent two years was evidently lower than those in the early 1970s and 1990s (P < 0.01) while the ratio of staphylococcus aureus was much higher than those in the middle of 1980s and 1990s (P < 0.01), in which methacillin -- resistant Staphylococcus aureus (MRSA) accounted for 85% or so, which increased significantly than those in the middle of 1980s and the beginning of 1990s (P < 0.01) with an increasing tendency. CONCLUSION: The increasing tendency of Staphylococcus aureus in our unit for the recent two years might closely be related to the preferential application of Imipenem in burn infection control. Furthermore, the ratio of MRSA in Staphylococcus aureus could not be lowered by vancomycin. PMID- 11876916 TI - [Analysis of bacterial resistance to antibiotics in a burn ward]. AB - OBJECTIVE: To analyze the bacterial resistance to antibiotics in a burn ward in the past 4 years. METHODS: Six hundred and fifty strains of bacteria were isolated from the wounds and blood of burn patients admitted to our hospital in recent 4 years. Slip agar diffusion method was employed to carry out antibiotic sensitivity test, and the results were interpreted according to the NCCLS criteria. Extended spectrum beta -- lactamases (ESBLs) were detected by double slips synergic test in 45 strains of Klebsiella Pneumoniae. RESULTS: Only vancomycin was effective for staphylococcus aureus, and Imipenem was effective for E. coli, E. cloacae and K. pneumoniae. But P. aeruginosa was only susceptible to polymyxin -- B. In addition, Thirty -- eight out of 45 strains of K. pneumoniae were ESBLs positive with the incidence of 84.4%. CONCLUSION: The high resistance of the bacteria to most antibiotics in our burn ward might be related to the high production of beta -- lactamases and especially ESBLs. Imipenem remained stable to ESBLs and remained highly effective for most bacteria. But antibiotics with beta -- lactamases inhibitor became less effective in recent years. The resistance rate of P. aeruginosa to Imipenem increased gradually in recent years. This might be due to the abuse of Tienam in burn patients, which stimulated the bacteria to produce a new metallic beta -- lactamases transfected by plasmid, decreasing the production of bacterial outer membrane proteins and inducing a change in membranous permeability. PMID- 11876917 TI - [A study of methicillin -- resistant staphylococcus aureus (MRSA) in a burn unit with repetitive -- DNA -- sequence -- based PCR fingerprinting]. AB - OBJECTIVE: To investigate the distribution and spread of MRSA in a burn ward, so as to explore the measures of the prevention, surveillance and control of hospital infection in a burn ward. METHODS: Five hundred and four specimens were isolated from the wounds and nasal vestibules of burn patients, the hands and nasal vestibules of medical staffs and lay attendants and the surfaces of various equipments. From these specimens, 58 strains of MRSA and 43 methicillin -- sensitive staphylococcus aureus (MSSA) were isolated. The genome DNA of isolated MRSA strains was analyzed by repetitive DNA -- sequence -- based PCR analysis. RESULTS: MRSA strains were isolated from the burn wounds in 22 of 41 (53.7%) patients, and 5 from the nasal vestibules. Moreover, among 19 medical staffs, MRSA strains were isolated from the hands of 9 persons, but not from the nasal vestibules. From the hands in 9 of 43 lay attendants and the nasal vestibules in 2 MRSA strains were found. Thirteen MRSA strains were isolated from 193 specimens from the surrounding items. It was indicated by repetitive DNA -- sequence -- based PCR analysis of the genome DNA of isolated MRSA strains that there existed homologous strains around the patients' wounds, in the burn wounds, on the skin of medical staffs and lay attendants and also surrounding equipments. CONCLUSION: (1) There was wide spread presence of MRSA homologous strains in the burn ward. (2) It was indicated by repetitive DNA -- sequence -- based PCR analysis of the genome DNA of isolated MRSA strains that there was cross infection among burn patients. The source of the infection of MRSA in burn ward was burn patients, and the route of the infection was hands of medical staffs and lay attendants. (3) MRSA is wide -- spread. The contamination of the hands and the environment was potential risk factor of MRSA outbreak in the burn unit. PMID- 11876918 TI - [The change in resistance of Pseudomonas aeruginosa to gentamicin during in vitro subculturing]. AB - OBJECTIVE: To observe the change in resistance of Pseudomonas aeruginosa (PA) to gentamicin (GM) during in vitro subculturing. METHODS: Twenty -- six strains of PA with resistance to GM isolated from burn patients were subcultured in vitro for 30 generations. The strains were also divided into high level (20 strains) and medium level (6 strains) of resistant groups according to their MIC (minimum inhibition concentration) values. The MIC and MBC (minimal bactericidal concentration) levels were examined in every strains of the 1st, 10th, 20th, and 30th filial generations of PA strain. RESULTS: There was no significant general difference of MIC and MBC values among the filial generations in each group (P < 0.05) when multiple comparisons of MIC and MBC mean values were made among every filial generations in high and medium levels of GM resistant groups. CONCLUSION: There was no evident difference in GM resistance among all the filial generations of PA. The result indicated that, the PA with different levels of GM resistance clinically found might come from different PA strains. PMID- 11876919 TI - [42 years' experience on the prevention and treatment of systemic infection after severe burn]. AB - OBJECTIVE: To summarize the experience of the management of 329 patients with major burn accompanied by severe systemic infection during the past 42 years, so as to help improve the treatment standard. METHODS: One thousand and one hundred twenty seven cases with major burns (TBSA > 50%) were admitted to our hospital during the past 42 years. The incidence of postburn systemic infection and change in the mortality were analyzed in terms of 3 periods, i.e. 1958 to 1980, 1981 to 1990 and 1991 to 2000. RESULTS: Postburn systemic infection developed in 329 out of 1127 cases with the morbidity of 29.4%. The incidences of the infection were 48.5%, 29.3% and 13.1% in the 3 periods, consecutively, with significant differences between them (P < 0.01 similar 0.05). During the 3 periods of past 42 years, the overall cure rate and cure rates in patients with different TBSA increased gradually, especially in those with TBSA Larger than 70%. CONCLUSION: The management of pastburn systemic infection was satisfactorily achieved by early comprehensive clinical measures such as immediate fluid resuscitation, early feeding, appropriate application of antibiotics and early escharectomy en masse, etc. which were established on the basis of experimental results. PMID- 11876920 TI - [The effects of extensive excision of massive invasive infected burn wound on the REE of burn patients with sepsis]. AB - OBJECTIVE: To explore the effects of extensive excision of massive invasive infected burn wound on the REE of burn patients with sepsis. METHODS: REEs and plasma levels of IL -- 6, IL -- 8, TNFalpha and LPS were determined before and after surgical interventions and when patients', condition improved in 8 burned cases with sepsis. RESULTS: All the 8 patients survived after treatment. The REE level in patients after operation was significantly lower than those before the operation (P < 0.01). REE in patients whose condition improved decreased obviously when compared to that after operation (P < 0.01). The plasma levels of IL -- 6, IL -- 8, TNFalpha and LPS decreased markedly after the operation when compared to those before operation (P < 0.05). and there were lower levels of these factors when the patients', condition improved (P < 0.001). Furthermore, there were closely positive correlations between REE and plasma IL -- 6, IL -- 8, TNFalpha and LPS (P < 0.01). CONCLUSION: Over -- releasing of some inflammatory mediators could be corrected by means of extensive excision of massive invasively infected burn wound. This might be beneficial to the control or the amelioration of the hypermetabolism in burn patients with sepsis. PMID- 11876921 TI - [Clinical study on the early and short -- term use of antibiotics with broad spectrum in severely burned patients]. AB - OBJECTIVE: To investigate the prevention and treatment effects of early and short -- term use of antibiotics with broad spectrum on postburn severe infection in severely burned patients. METHODS: Thirty -- five burn patients with TBSA from 50% to 95% were enrolled in the study. The patients were divided into early prevention [20 cases, antibiotics were used started from 6 postburn hours (PBH)] and delayed prevention (15 cases, antibiotics were applied after 48 PBH) groups. Plasma levels of LPS, TNFa and IL -- 8 were dynamically monitored with the concomitant observation of clinical signs of postburn sepsis. RESULTS: After major burns, the plasma levels of LPS, TNFalpha and IL -- 8 increased evidently and reached the peak values on 3 similar 5 postburn days (PBD). But the levels of all above factors in the early group were obviously lower than those in the delayed group (P < 0.05 similar 0.01). The incidencies of sepsis and internal organ complications within 2 postburn weeks were much lower in the early group than those in the delayed group (P < 0.05). The subeschar bacterial quantification on 4 similar 7 PBD was evidently lower in the early group than that in the delayed group (P < 0.01). CONCLUSION: Early and short -- term use of antibiotics with broad spectrum in severely burned patients could effectively prevent postburn severe infection and lower down the incidence of internal organ complications. PMID- 11876923 TI - [Clinical investigation of burn wound infection with type I herpes simplex virus]. AB - OBJECTIVE: To elucidate the clinical features of burn wound infection with type I herpes simplex virus (HSV -- 1), so as to propose the possible therapeutic measures. METHODS: Burn wound infection with HSV -- 1 was diagnosed in 7 cases by means of bacterial culture, PCR detection of virus, immunohistochemical staining and pathomorphological section with the exclusion of other causes. In addition, the clinical signs and the features of the development of the infection in 62 cases were recorded and analyzed. RESULTS: In the burn patients with the infection, the burn wounds were covered by tumor-like masses with the cauliflower appearance. In the 7 cases who underwent pathomorphological examination, 5 with positive HSV -- I. Epithelial cells exhibited balloon -- like appearance, and reticular degeneration, and inclusion bodies were found in balloon-like cells under microscope examination. In the late stage, there revealed obvious proliferation of epithelial prickle cell layer in shape of pseudopapillomatous mass with the obvious liquefaction of basal cells. The sufferers exhibited evident local signs with mild systemic reaction. The management of the wound included wet compress with the addition of neomycin and pentacyclic guanosine. CONCLUSION: With the employment of PCR virus detection and immunohistochemical staining, burn wound infection with HSV -- I could be identified. Non -- surgical treatment could be effective. PMID- 11876922 TI - [A study on bactericidal/permeability increasing protein (BPI) as a natural inhibitor of endotoxin]. AB - OBJECTIVE: To investigate the existential status of BPI as a natural defensive factor in human polymorphonuclear leukocytes (PMNL). METHODS: The abilities of porcine BPI to combine endotoxin (lipopolysaccharide, LPS) and inhibit the release of TNFalpha from hepatic Kupffer's cells were examined. And the expression of BPI in PMNL after stimulation by LPS was determined by flow cytometry. RESULTS: The ability of porcine BPI to combine LPS was increased along with the increment of BPI concentration. The effects of the BPI on the inhibition of the release of TNFalpha from hepatic Kupffer's cells were dose-dependant. The expression of BPI in PMNL was enhanced obviously within 30 mins of LPS stimulation. But there was no immediate release of BPI into blood. CONCLUSION: BPI possessed potential power of neutralizing LPS. It might be beneficial to supplement exogenous BPI in case of Gram negative bacterial infection. PMID- 11876924 TI - [An experimental study on the construction of composite graft with epithelia and fibroblasts on collagen sponge]. AB - OBJECTIVE: Primarily cultured human epithelia and fibroblasts were inoculated on collagen sponge to construct composite skin substitute. METHODS: Epithelia and fibroblast asts from human foreskin were primarily cultured. Fibroblasts (1 x 10(5)/cm(2)) were inoculated on collagen sponge after digestion and cultured for 3 days prior to inoculation of epithelial cells (2 x 10(5)/cm(2)) to construct composite skin grafting substitute, and it was then grafted onto full skin loss (FSL) wounds of nude mice. Collagen sponge membrane lacking cell inoculation was used as control. The wound healing was evaluated by measuring the residual wound area. Tissue samples were harvested and examined by means of histology, immunohistochemistry and electron microscopy. RESULTS: Epithelia and fibroblasts could be cultured in vitro on the carrier collagen sponge. After being grafted onto the wound, the epithelia and fibroblasts wound further proliferated and differentiated. The composite skin could stick to the carrier fairly well. There exhibited less epidermal scar proliferation and satisfactory contour of the neoskin. Integral basal membrane of the skin could be identified by immunohistochemical staining. The quality and speed of wound healing with the composite skin substitute were superior to that with collagen sponge only. CONCLUSION: The composite grafts constructed by epithelia and fibroblasts on collagen sponge could be a potential neotype composite skin substitute for the repair of FSL. PMID- 11876925 TI - [An Experimental Study on the diagnosis of high voltage electrical injury by means of biological oxygen consumption measurement]. AB - OBJECTIVE: To investigate the degrees of the degeneration or the necrosis of muscle tissue injured by high voltage electricity as determined by biological oxygen consumption measurement. METHODS: An animal model with high voltage electricity designed in our laboratory was employed in the study. Muscle tissue sized 12 mm x 10 mm x 1 mm at electric burn produced 5 spots of the rabbit injured legs was harvested for the measurement of tissue oxygen consumption for 100s at 2, 8, 14, 24, 48 postburn hours and 5 postburn day. At the same time histological examination was made. RESULTS: One hundred and eighty two pieces of tissue samples were examined. Among them 20 samples with oxygen consumption below 8.7 kpa/100s (43.5%) showed necrosis under LM, while 157 samples with oxygen consumption of 16.9 to 8.78 kpa/100s (84% similar 44%) showed histological necrosis and different degrees of mixed degeneration and normal tissue, and 5 samples with oxygen consumption above 17 kpa/100s (85%) showed normal histological structure. CONCLUSION: The lower the tissue oxygen consumption was, the more severe the tissue degeneration and necrosis were. Oxygen consumption measurement could be valuable in the determination of the degree of tissue injury inflicted by electricity in terms of its convenience, rapidity and accuracy. PMID- 11876926 TI - [An experimental study on the relationship between early myocardial injury and the in situ expression of TNFalpha mRNA in burned rats with endotoxemia]. AB - OBJECTIVE: To observe the in situ expression of TNF mRNA and protein in myocardial cell, so as to explore the possible mechanism of early myocardial injury in scalded rats with endotoxemia. METHODS: One hundred and seventy eight Wistar rats inflicted with 20% TBSA III degree napalm burn compound with endotoxemia and multiple organ injury were employed as the model and were randomly divided into four groups as follow. (1) burn and infusion group (BI): the rats were burned and infused with endotoxin. (2) simple burn group (S): the rats were burned only. (3) simple endotoxin infusion group (EI): the rats were only infused with endotoxin. (4) normal control (C). The observation time points were designed to be 0.5, 1, 3, 6, 12, 24 and 48 postburn hours (PBH). The morphological and functional changes in rat myocardium, the change in serum TNFalpha, the cellular location and distribution of TNFalpha mRNA and proteins were observed by light microscope (LM), electronic microscope (EM), ELISA, immunohistochemistry and in situ hybridization staining. RESULTS: There exhibited a series of pathomorphological changes in the myocardium at early postburn stage in rats of BI group, such as wavy degeneration of myocardial fibers, the formation of contractile bands, fragmentation of myocardial fibers and focal cytoplasmic lysis, etc. The left ventricular systolic pressure (LVSP) and the maximum change rate of intraventricular pressure (plus minus dp/dtmax) decreased significantly (P < 0.01). Serum TNFalpha level increased obviously at 1 PBH (P < 0.01) and peaked at 3 similar 6 PBH. It was found that TNFalpha mRNA was principally located in myocardial cells and in a part of inflammatory cells. But these milder pathomorphological changes in myocardial injury were found in rats of S and EI groups. In addition, there appeared elapsed TNFalpha peak values and relative weaker expression of myocardial TNFalpha mRNA. CONCLUSION: TNFalpha mRNA and protein could be expressed in myocardium in burned rats with endotoxemia which might be one of the origins of TNF in participating in the development of early postburn myocardial structural and functional injury. PMID- 11876927 TI - [Enteral feeding of glucose increases intestinal mucosal blood flow during intestinal ischemia/reperfusion injury]. AB - OBJECTIVE: To explore the relationship between enteral nutrition and the change in intestinal mucosal blood flow during intestinal ischemia/reperfusion injury. METHODS: Thirty-six Sprague-Dawley rats were randomly divided into alanine (12 rats), glucose (14 rats) and mannitol control (10 rats) groups. Jejunal sac was prepared with the filling of either 10 mM alanine, glucose or mannitol in the three groups. The laser doppler probe and intestinal mucosal tonometry were placed at the both ends of the sac. The superior mesenteric artery was occluded by arterial clamp for 60 mins and released thereafter for another 60 mins. Intestinal mucosal blood flow and regional pressure of CO(2) (PrCO(2)) were determined every 30 mins. RESULTS: During the process of ischemia/reperfusion, the intestinal mucosal blood flow in glucose group increased evidently and the PrCO(2) in glucose group decreased obviously (P < 0.01) when comparcd with those in mannitol group. CONCLUSION: During the process of ischemia/reperfusion, enteral feeding of glucose could increase intestinal mucosal blood flow, which provided guarding effects on the intestine suffering from ischemic/reperfusion injury. PMID- 11876928 TI - [The predictive value of serum neopterin for multiple organ dysfunction syndrome in extensively burned patients]. AB - OBJECTIVE: To evaluate the predictive value of serum neopterin for multiple organ dysfunction syndrome (MODS) in extensively burned patients. METHODS: Twenty-nine burn patients with TBSA of more than 70% were included in this study, and these patients were divided into two groups according to the occurrence of MODS. Blood samples were obtained on 1, 3, 7, 14, 21 and 28 postburn days (PBDs). And serum neopterin levels were determined by radioimmunossay. To evaluate the predictive power of neopterin, sensitivity, specificity, predictive value of a positive/negative test were calculated for the diagnosis of MODS. RESULTS: Serum neopterin levels in MODS group were significantly higher than those in non-MODS group on 3, 7, 14 and 21 PBDs (P < 0.05 similar 0.01). And a positive correlation was found between neopterin levels and MODS scores in patients who developed MODS on 3, 7 and 21 PBDs (r = 0.470 similar 0.811). In a period of up to 28 PBDs, neopterin levels exceeding 35.0 nmol/L were associated with MODS by a clinical sensitivity of 81.1%, a specificity of 89.9%, a positive predictive value of 81.1%, and a negative predictive value of 89.9%. CONCLUSION: These data indicated that the presence of constant high neopterin serum levels appear to be associated with the development of postburn MODS. Serum levels of neopterin might be a useful marker for the early identification of extensive burned patients at risk for MODS. PMID- 11876929 TI - [The dynamic changes of serum TNFalpha level in scalded rats after the administration of rhGH during the early postburn stage]. AB - OBJECTIVE: To examine the dynamic change of serum TNFalpha level in scalded rats after the administration of rhGH during the early postburn stage. METHODS: Forty two male sanitary Sprague-Dawley rats were randomly divided into three groups, i.e. normal control (N), burn with early administration of rhGH (BA) and burn without rhGH (BN) groups. The rats were inflicted with 25% similar 30% TBSA deep partial thickness scalding on the back. rhGH in dose of 6 U/kg was administered subcutaneously via abdominal skin starting from the 1st postburn day (PBD). Rat serum TNFalpha levels were determined by ELISA in BA and BN groups at 3, 6 and 10 PBDs. RESULTS: The rat serum TNFalpha level in BA was lower than that in BN groups on 3 PBD. Whilst the TNFalpha levels in BA were much higher than those in BN groups on 6 and 10 PBDs. CONCLUSION: Administration of rhGH at early postburn stage might abate the early systemic postburn reaction. PMID- 11876931 TI - [The effects of rhGH on macrophages in scalded mice]. AB - OBJECTIVE: To investigate the effects of rhGH and IGF-1 on the CD14 mRNA expression and cytokine secretion of peritoneal macrophages (Mphi) in scalded rats. METHODS: Reverse-transcription-polymerase chain reaction (RT-PCR) and enzyme-linked immunosordent assay (ELISA) were employed to observe the postburn changes of CD14 expression and cytokines secretion of the Mphi treated in vivo and in vitro by rhGH and IGF-1. RESULTS: It was demonstrated that in vivo postburn application of rhGH and IGF-1 could enhance the transcription of CD14 mRNA and the secretion of cytokines from the peritoneal Mphi in mice. But it seemed that there was no additive effect between the two factors. Additionally, rhGH (at 40 ng/ml) and IGF-1 (at all tested concentrations) could activate the secretion of TNF and IL-6 from cultured mice Mphi in vitro. CONCLUSION: Postburn application of rhGH might activate Mphi by IGF-1 induction, enhancing Mphi CD14 expression, which promoted more secretion of cytokines, and promote the enhancement of immune function. PMID- 11876930 TI - [An experimental study on systemic inflammatory response syndrome induced by subeschar tissue fluid]. AB - OBJECTIVE: To investigate the effects of subeschar tissue fluid (STF) on normal rats and their peritoneal macrophages in vitro, so as to explore its relationship with inflammatory mediators. METHODS: Eighteen Wistar rats were evenly and randomly divided into three groups, i.e. STF, normal human serum control and blank control groups. The changes of the heart and respiratory rates of rats in every group were recorded by two channel physiological recorder. Blood samples were collected for the determination of hepatic and renal function. The hepatic, pulmonary, renal and cardiac tissues were harvested for routine morphological examination. The changes of the secretion of TNFalpha, IL-1 were also determined. RESULTS: The heart and respiratory rates increased after the infusion of STF, and leukocyte count increased along with different degrees of injury to lungs, liver and kidneys. It was indicated in in vitro experiment that STF could activate macrophage and stimulate the secretion of TNFalpha and IL-1 from the macrophage. CONCLUSION: STF might induce the development of SIRS in rats, which further lead to the development of MODS. PMID- 11876932 TI - [The effect of Lipopolysacharide (LPS) on morphology and function of human umbilical endothelial cells (HUVECs)]. AB - OBJECTIVE: To study the effects of LPS on the morphology and function of HUVECs, so as to explore the roles of activated vascular endothelial cells (VEC) by LPS in systemic inflammatory reaction, sepsis and MOF. METHODS: HUVECs primarily cultured in vitro were employed as the model. Inverted microscope was used to observe the effects of LPS on the morphology of HUVECs. ELISA was used to assess the IL-6 content. The expression of ICAM-1 was determined by immunofluorescent staining method with confocal laser-scanning fluorescence microscope. RESULTS: The morphology of HUVECs could be altered by LPS to become long cord-like in shape. The smallest dose of LPS for stimulating the secretion of IL-6 from HUVEC was 1 ng/ml, and the effect was enhanced parallel to the increase of the concentration of LPS and reached top level at 8 postburn hours. The expresson of ICAM-1 on the nucleic and cytoplasmic membrane of HUVECs increased obviously after HUVECs was cultured with LPS (10 ng/ml) for 24 hours when observed by laser confocal scanning light microscope. CONCLUSION: The morphology and function of HUVECs could be evidently affected by LPS. As a result, LPS might play important roles in the increase of vascular permeability, the promotion of leukocytic adherence, and the initiation of inflammatory cascade reaction. PMID- 11876933 TI - [A study on the effects of CD3AK cells on the improvement of cellular immune function in burned patients]. AB - OBJECTIVE: To explore new ways of correcting postburn immune suppression in severely burned patients. METHODS: CD3AK cells were cultured in serum-free medium and were employed to treat patients with major burns. Similar patients treated by routine treatment were set to be control group. The changes of cellular immunological indices were dynamically observed in the patients of two groups at different time points. RESULTS: Compared to those in control group, the ratio of CD4+ cell of T cell subgroup increased in patients treated by CD3AK, in concomitant with the decrease of the ratio of CD8+ cells and the increase of the ratio of CD4/CD8. Serum soluble IL-2R level decreased. T lymphocyte transforming activity increased and NK cell activity increased. CONCLUSION: CD3AK cells cultured in serum-free medium could improve and restore the decreased cellular immune function in burn patients. PMID- 11876934 TI - [The effects of astragalus and shenmai injections on macrophage function in burned mice]. AB - OBJECTIVE: To explore the dynamic postburn change in macrophage function in burned mice within 120 hrs after injury, and to investigate the effects of astragalus and shenmai injections on the macrophage function and surrvival rate of burned mice. METHODS: The mice were divided into 13 groups according to postburn time and handling methods, i,e, normal control (A), burn control (B), normal mice with astragalus (NA), normal mice with shenmai (NS), burned mice with astragal (BA), burned mice with shenmai (BS) 2 postburn hour (2 PBH), 6 PBH, 12 PBH, 24 PBH, 48 PBH, 72 PBH, 120 PBH groups. The changes in the various macrophage functions at different postburn time points and after the use of astragalus and shenmai injections were determined by means of phagocytic and RT PCR methods. RESULTS: (1) Within 120 PBHs, the phagocytic function of murine macrophages decreased evidently. The ACP activity decreased obviously. The expression of IL-15 mRNA fluctuated and that of TNF mRNA enhanced significantly. (2) Five days after the application of astragalus in dose of 2 500 mg . kg(-1) .d(-1), the phagocytic function of macrophages and ACP activity increased markedly (P < 0.01). The expressions of IL-15 and TNF mRNAs were not influenced. The survival rate of mice was not increased. (3) Five days after the application of shenmai injection in dose of 2.5 ml . kg(-1) . d(-1), the phagocytic function of macrophages and ACP activity increased significantly (P < 0.01), while the expression of IL-15 mRNA exhibited no change. But the expression of TNFalpha mRNA decreased obviously (P < 0.01). Moreover, the survival rate of burned mice was evidently raised (P < 0.05). CONCLUSION: Peritoneal administration of shenmai injection at early postburn stage could significantly improve the macrophage function of burned mice, and it increase the survival rate of mice. PMID- 11876935 TI - [A study on the direct binding of LPS to endothelium and the characteristics of endothelial injury]. AB - OBJECTIVE: To explore the characteristics of the combination of LPS to the endothelium and endothelial injury. METHODS: Myocardial microvascular endothelial cells (MMECs) were isolated and cultured to adhering state. Flow cytometry, laser confocal scanning microscope (LCSM) and image analysis were employed to evaluate the effects of different concentrations of LPS and different culturing time with the same concentration of LPS on the adhered MMECs RESULTS: (1) The OD values of MMECs increased progressively along with the prolongation of culturing time of MMECs with LPS and reached top level at 30 min. In addition, the increase of OD values was dependent on time. (2) The number of positive MMECs and the OD values increased after MMECs was cultured for 2 hours with LPS at concentrations ranging from 0.03125 similar 2.0000 g/L. The OD value reached peak level when LPS concentration was 0.2500 g/L, which implied that this concentration of LPS (0.25 g/L) was the optimal one for the combination of LPS with MMECs, and there was concentration saturation. (3) LPS could enter into the cytoplasm and nucleus of MMECs. (4) LPS could induce nucleic translocation and denucleation. CONCLUSION: (1) LPS could combine MMECs and enter into the nucleus without the presence of serum. (2) LPS could directly damage MMECs. PMID- 11876936 TI - [An experimental study on the apoptosis of PMNs and macrophages during the early postburn stage in severely scalded rats]. AB - OBJECTIVE: To investigate the changes of postburn apoptosis of PMN and the apoptosis of PMN and macrophages after being stimulated by burn sera and the change of the phagocytosis of apoptotic PMNs by macrophages. METHODS: Rats inflicted by 30% TBSA III degree scalding were employed as the model. PMNs were harvested on the 0, 6, 12 and 24 postburn hour (PBH). Burn sera were harvested at 24 PBHs. PMN apoptosis was detected by flow cytometry. Simultaneously, peritoneal macrophages were collected for the study. The apoptotic change of the macrophages (Mphis) after being stimulated by burn sera and the ability of Mphis phagocytizing apoptotic PMNs at 24 PBHs were examined. RESULTS: The PMN apoptosis was obviously inhibited at 6, 12 and 24 PBHs in the scalded rats with no difference between the time spots. Twenty-four hours after being stimulated by burn sera, the apoptotic rate of PMNs on scalded rats was lower than that in normal rats, while the apoptotic rate of Mphis in scalded rats was higher than that in normal rats. The ability of Mphis phagocytizing apoptotic PMNs decreased after severe burn (P < 0.01). CONCLUSION: There exhibited delayed rat PMN apoptosis and decreased ability of the phagocytosis of apoptotic PMNs by the Mphis after severe burn or being stimulated by burn sera. This might lead to the incomplete phagocytosis of apoptotic PMNs by Mphis. As a result, the PMNs would finally come to necrosis and release toxic contents which were important factors in the inducing of SIRS. PMID- 11876937 TI - [The effect of heparin on the deep second-degree burn in scalded rats]. AB - OBJECTIVE: To investigate the effects of heparin on deep second-degree burn in scalded rats. METHODS: Thirty Sprague-Dawley rats inflicted with 20% TBSA deep second-degree scalding on the back were employed as the model and were randomly divided into two groups. i,e heparin (H) and normal saline groups (N). Heparin (100 U . kg(-1) . day(-1)) was injected subcutaneously to the rats in H group while normal saline to those in N group once a day. Wound healing rate and immunological indices were determined simultaneously. RESULTS: (1) The wound healing time was much shorter in H group than that in N group (P < 0.005). The granulation tissue growth condition examined by LM and EM was much better in H group that than in N group. (2) The lymphocytic transforming ratio in H group was evidently increased (P < 0.005). And the phagocytic function of the macrophages increased significantly (P < 0.05). But there was no difference between the two groups in terms of hemoltytic plaque test (P > 0.05). CONCLUSION: Injection of heparin subcutaneously could accelerate burn wound healing and strengthen the systemic cellular immune potential and the macrophage phagocytic capacity of the host, but exerted no effect on the humoral immunity. PMID- 11876938 TI - [The Influence of stress inhibition on the plasma levels of LPS, pro-inflammatory and Th1/Th2 cytokines in severely scalded rats]. AB - OBJECTIVE: To explore the influence of stress inhibition on the plasma levels of LPS pro-inflammatory and Th1/Th2 cytokines in severely scalded rats. METHODS: Sprague-Dawley (SD) rats inflicted by 30% TBSA of deep partial thickness burn were employed as the model and randomly divided into burn with immediate resuscitation (A) and burn with immediate resuscitation and soluble cocktail (B) groups. Plasma was harvested from peripheral blood at different postburn time points for the determination of the levels of LPS, IL-1alpha, IL-6, TNFalpha, IL 8, IL-2, IFN-gamma, IL-4 and IL-10 in the rats of the two groups. And the rats inflicted by sham scalding were taken as control group (C). RESULTS: The postburn plasma levels of LPS, IL-1alpha, IL-4 and IL-10 increased gradually, while the plasma levels of IL-6, TNF, IL-8, IL-2 and IFN increased initially and decreased thereafter. The increasing ranges of LPS and these inflammatory cytokines were higher in A group, in which the increases of IL-1alpha, IL-6 and IL-4 appeared earlier in A (6 PBH) than those in B (12 PBH) groups. CONCLUSION: Prompt fluid resuscitation and stress inhibition could delay and ameliorate the postburn inflammatory reaction, decrease the production of Th2 cytokine and partially restore the production of Th1 cytokine after 48 PBH. PMID- 11876939 TI - [The influence of recombinant human growth hormone on the metabolism of glucose and protein in severely burned patients]. AB - OBJECTIVE: To investigate the influence of recombinant human growth hormone (rhGH) on the metabolism of glucose and protein in severely burned patients. METHODS: The serum levels of total protein and albumin and the incidence of hyperglycemia in 24 cases of severely burned patients treated by rhGH were determined and compared with those in patients without rhGH treatment (control group). RESULTS: The serum levels of total protein (TP) and albumin in burned patients after being treated by rhGH were much higher than those before treatment and those in control group. The blood level of glucose increased within 2 weeks, especially 1 week postburn. The incidences of hyperglycemia were 52.38%, 50.24% and 20% on 0 similar 3, 4 similar 7 and 8 similar 14 postburn days (PBDs), respectively. And the incidences of hyperglycemia were 100%, 50% and 0% when the administration of rhGH started within 1, 2 and after 2 weeks respectively. CONCLUSION: The systemic protein synthesis in severely burned patients could be accelerated by the use of rhGH. But early administration of rhGH in burned patients could affect blood sugar level obviously. And the optimal time to give rhGH is 2 weeks after burn injury. PMID- 11876940 TI - [The relationship between postburn enterogenic hypermetabolism and decontamination of intestine]. AB - OBJECTIVE: To explore whether postburn enterogenic hypermetabolism exists. METHODS: Eighty -- eight Wistar rats inflicted by 30% TBSA III degree burns were randomly divided into two groups, i,e. burn (B) and burn with gut decontamination (D) groups. The change in resting energy expenditure (REE) was observed during 0 to 10 postburn day (PBDs) in rats. The plasma contents of lipopolysaccharide (LPS), tumor necrosis factor (TNF) and interleukin-1 (IL-1) were determined on 1, 3, 5, 7 and 10 PBDs and their correlations were analyzed. RESULTS: The postburn REE and plasma contents of TNF, LPS and IL-1 were much higher in both groups than those before injury. When compared between the two groups, the above data in D group were lower in varying degrees than those in B group. It was shown by correlation analysis that REE was postively correlated to LPS, TNF and IL-1, respectively (gamma = 0.77 similar 0.9, P < 0.05 similar 0.01). CONCLUSION: Postburn hypermetabolism in rats could be ameliorated by gut decontamination, and the postburn plasma inflammatory mediators in rats could also be lowered by gut decontmination. This suggests that enterogenic hypermetabolism does exist after burn injury. PMID- 11876941 TI - [Clinical observation of the protective effect of oral feeding of glutamine granules on intestinal mucous membrane]. AB - OBJECTIVE: To observe the protective effect of oral feeding of glutamine (Gln) granules on intestinal mucous membrane during early postburn stage. METHODS: Twenty burn patients were randomly divided into test (T) and control (G) groups with 10 patients in each group. Gln granules and placebo were administered in a dose of 10 grams three times a day for 10 days to patients in the two groups respectively. Blood levels of Gln, diamine oxidase (DAO) activity, endotoxin (LPS) and intestinal permeability were monitored before and after the administration of the agents. at the same time, the changes in blood and urine routine and hepatic and renal functional indices were also observed. RESULTS: There was no difference of the general data and the hepatic and renal functional indices between the two groups. The plasma level of Gln before the administration was much lower than that thereafter (P < 0.01) in T group, but all of the values were higher than that in C group (P < 0.05). The DAO activity, intestinal permeability and LPS levels before the administration were much higher than those after it (P < 0.01) in T group. CONCLUSION: Gln might be good to burn patients due to its protective effect on intestinal mucous membrane when given during early postburn stage. PMID- 11876942 TI - [An experimental study on the effects of early postburn nutrition on calcitonin gene--related peptide]. AB - OBJECTIVE: To explore the resuscitation effects of different nutritional routes during early postburn stage on intestinal tract, and the relationship between calcitonin gene -- related peptide (CGRP) and resuscitation of intestinal tract. METHODS: Sixty Wistar rats inflicted by 30% TBSA III degree flame burn were randomly divided into burn with enteral feeding group (EF, 30 rats) and burn with early parenteral nutrition group (EPN group, 30 rats). Six normal rats without burn injury served as control group (C). The changes in general condition, intestinal mucosal blood flow, plasma concentration of CGRP immunologic competence substance and intestinal CGRP positive staining substance in the rats were observed at 6, 12, 24, 48 and 72 postburn hours (PBHs). RESULTS: The rat intestinal mucosal blood flow decreased postburn, especially in EPN group (P < 0.05). The postburn plasma CGRP increased and the plasma CGRP level in EF group was obviously lower than that in EPN group at 72 PBH (P < 0.01). The CGRP positive staining cells of intestinal tract decreased markedly postburn (P < 0.05) in both burn groups, and those in EPN were significantly lower abundant than those in EF group at 24 and 48 PBHs (P < 0.05). Furthermore, the decrease in intestinal mucosal blood flow was positively related to the decrease in intestinal intramuscular nerve plexus CGRP (r = 0.72, P < 0.05). CONCLUSION: Enteral nutrition exerted better effects on early postburn intestinal resuscitation than parenteral nutrition. The decrease in intestinal blood supply might be mainly due to the excessive increase in plasma CGRP immune active substance and the decrease of postburn intestinal tract CGRP immunologic competence substance. Early enteral feeding might play resuscitative role by modulating CGRP. PMID- 11876943 TI - [The change in plasma concentration of free amino acids during early postburn stage in severely scalded rats]. AB - OBJECTIVE: To investigate the rule of the change in plasma concentration of free amino acids during early postburn stage in severely scalded rats, so as to provide assistance for the study of postburn abnormal metabolism of skeletal muscle. METHODS: Male Wistar rats inflicted by 30% TBSA III degree scalding were employed as the model and were randomly divided into normal (C), 2 postburn hour (PBH), 6PBH, 12PBH and 24PBH groups, with 8 rats in each group. Plasma concentrations of free amino acids was determined by automatic amino acid analyzer. At the same time, the plasma contents of alanine aminotransferase (ALT), aspartate aminotransferase (AST), cortisol, TNF and IL-6 were also assayed. RESULTS: There was adecreasing tendency in varying degrees in plasma concentration of all amino acids at each time points within 24 PBHs although there was no obvious difference in total amino acids content at each time point. The plasma level of BCAA decreased evidently at 2 PBH and increased obviously at 12 PBH, while the plasma levels of aromatic amino acids (AAA) increased evidently at 12 and 24 PBHs. In addition, the BCAA/AAA ratio showed no change. The ratio of phenylalanine to tyrosine increased significantly at all postburn time points except that of 2PBH. The plasma levels of ALT, AST increased obviously at all postburn time points (P < 0.01). Plasma cortisol levels in all scalding groups were markedly higher than that in C group (P < 0.01), and reached peak level at 12 PBH. Plasma contents of TNFalpha and IL-6 increased significantly (P < 0.01). Whereas the changes in IL-6 and cortisol were obviously and positively correlated to the changes in 3 -- mH and Phe/Tyr. CONCLUSION: There were significant changes in plasma contents of free amino acids during early postburn stage in scalded rats, which might be related to the early excessive release of inflammatory mediators and enhanced degradation of skeletal muscle and impairment of hepatic function. The mechanism of the change needs to be further studied. PMID- 11876944 TI - [An experimental study of the effects of early enteral feeding of L--arginine enriched food on postburn intestinal mucosal proliferation in rats]. AB - OBJECTIVE: To investigate the effects of early enteral feeding of L -- arginine enriched food on postburn intestinal mucosal proliferation in rats. METHODS: Twenty Wistar rats prepared with gastrostomy 7 days before the experiment were inflicted by 30% TBSA flame burn, and were divided into early enteral feeding (EF), L -- arginine enriched early enteral feeding group (AEF) receiving feeding from 2 PBHs and delayed feeding (DF) group, receiving feeding from 48 PBHs. The changes in intestinal mucosal DNA content and intestinal mucosal proliferating cell nuclear antigen labeling index (PCNALI) were determined on the 1st, 3rd, 6th and 9th postburn days (PBDs) respectively in rats. RESULTS: There were rare intestinal mucosal PCNA positive cells on the 1st PBD in all groups. Compared with that in DF group, the intesinal mucosal PCNALI in AEF group increased obviously on 3 PBD, while that in EF group increased evidently on 6 PBD. (P < 0.05). CONCLUSION: There existed intestinal mucosal injury postburn as shown by the inhibition of intestinal epithelial proliferation. Early enteral feeding with L -- arginine enriched food could not only ameliorate intestinal mucosal injury, but also enhance the proliferation of intestinal mucosal epithelium. PMID- 11876945 TI - [Effects of tetrandrine on the synthesis of collagen and scar-derived fibroblast DNA]. AB - OBJECTIVE: To investigate the effects of tetrandrine on the synthesis of collagen and scar derived fibroblast DNA. METHODS: Scar-derived human fibroblasts were cultured in vitro. The changes in the levels of the synthesis of collagen and DNA of the fibroblasts were represented by the incorporation values of (3)H-TdR and (3)H-proline into the cells. Tetrandrine was added to the culture media of the cells, and its effects were studied. RESULTS: When the concentration of added tetrandrine increased from 5 mg/L to 80 mg/L, The (3)H-TdR values in scar-derived fibroblasts were 1162 plus minus 226 and 412 plus minus 82, respectively while that in control group was 1740 plus minus 165, showing an inhibition rate of 76.32%, and the difference (P < 0.01). In addition, the (3)H-proline incorporation values in the cells were 535 plus minus 141 and 341 plus minus 89, respectively, while that in control group was 1126 plus minus 193, with the inhibition rate of 69.71%, showing significant difference (P < 0.01). CONCLUSION: The synthesis of DNA and collagen in cultured scar-derived fibroblasts could be inhibited by tetrandrine in dose-dependent pattern. Tetrandrine might be a potential agent for the prevention and treatment of proliferative scars. PMID- 11876946 TI - [The adhesive and migrating function of human epithelial cells opsonized by fibronectin]. AB - OBJECTIVE: To investigate the adhesive and migrating function of human epithelial keratiocytes during in vitro culture. METHODS: The adhesive and migrating function was determined in freshly isolated EKCs and those cultured for 7 similar 10 days after being opsonized by fibronectin (FN). The expressions of alpha(5)beta(1) receptors in EKC were examined with indirect immunofluorescence staining methods before and after culture. RESULTS: (1) The adhesive and migrating indices after FN opsonization of EKCs freshly isolated EKCs were obviously lower than those cultured for 7 similar 10 days (P < 0.01). (2) There exhibited positive staining of the expression of alpha(5)beta(1) receptors in the EKC after 1 day culture and in the proliferative epithelia after in vitro culturing of a tissue mass. While there exhibited strongly positive staining of the peripheral proliferative epithelia of the EKCs and tissue mass after 7 days of culture, negative or weakly positive stainings were found in freshly isolated EKCs. CONCLUSION: (1) There existed significant difference of biological function between the freshly isolated EKCs and those cultured for 7 similar 10 days. (2) The strongest expression of alpha(5)beta(1) receptors was observed at the active proliferative site (peripheral proliferative epithelia of a tissue mass) and in the biological active period (cultured for 7 days) of EKCs. (3) The adhesive and migrating function of EKCs could be effectively induced and activated by in vitro culture, which enabled the EKC to alter from a relatively biologically functional static state to an actively functional state. PMID- 11876947 TI - [The effects of gut -- origin lipopolysaccharide translocation on the apoptosis of lymphocytes in scalded rats]. AB - OBJECTIVE: To investigate the relationship between lymphocyte apoptosis in spleen and mesenteric lymph nodes (MLNs) and gut -- origin lipopolysaccharide (LPS) translocation after severe scalding in rats. METHODS: Wistar rats inflicted 30% TBSA III degree scalding were employed as the model. Serum LPS levels in portal vein and vena cava were determined by limulus amebocyte lysate (LAL) test. Apoptotic lymphocytes in paraffin -- embedded spleen and MLNs sections were detected with TdT -- mediated and dUTP nick end labeling (TUNEL) and peroxidase (POD) staining. The images were taken by cooldccd camera system, and the count and optical density (OD) value of apoptotic lymphocytes were analyzed with software Spot and Imagine proplus 4.10a (IPP4.10a). RESULTS: Serum LPS levels in portal vein and vena cava increased sharply in rats postburn (P < 0.01) and reached a peak value at 6 postburn hour (PBH). LPS level in portal vein was higher than that in vena cava (P < 0.01) in the period of 3 to 48 PBHs. The results of TUNEL -- POD staining and the count of apoptotic lymphocytes showed that the apoptotic cells were few in normal MLNs, but increased obviously postburn (P < 0.01), especially at 3 PBH. On the contrary, apoptotic lymphocytes were concentrated in spleen cortex before injury and decreased obviously postburn (P < 0.01), though a slight recovery was found after 24 PBH. Optical density of apoptotic lymphocytes was significantly reduced in spleen at 24 PBH, But the OD value of apoptotic lymphocytes in MLNs increased significantly within 48 PBHs when compared with that before injury (P < 0.01). CONCLUSION: Gut -- origin endotoxemia in rat could occur in early postburn period. The LPS translocation might induce lymphocyte apoptosis in MLNs. The decreased apoptotic lymphocytes in spleen implied abnormal immune function in spleen. PMID- 11876948 TI - [Bioactivity and test grafting of acellular dermal matrix containing fibroblasts]. AB - OBJECTIVE: To investigate the bioactivity of acellular dermal matrix with fibroblasts and its role as dermal skeleton. METHODS: Human fibroblasts (HFs) were planted onto the surface of acellular dermal matrix (ADM) to form living dermal substitute. The IL -- 6, IL -- 8 and TGF contents in the supernatant of the culture of HF -- ADM were determined with ELISA method, and the secretion of hyaluronic acid and laminin from extracellular matrix was measured with RIA method. The speed of vascularization and the wound contracture rate were observed after the dermal substitute was grafted on the full skin loss wound of Balb/c-nu mice (nude mice). RESULTS: HFs grew very well after being planted onto ADM so as to form a single layer of cellular membrane. Many kinds of cytokines and extra cellular matrix components were secreted. Compared with simple acellular dermal grafting, the vascularization was accelerated, and the wound contracture rate decreased, after the living dermal substitute being grafted on the wound. CONCLUSION: The ADM seeded with HFs exhibited excellent bioactivity and might be an optimal dermal substitute. PMID- 11876949 TI - [The adherence of leukocytes mediated by L-selectin to endothelia activated by burn sera]. AB - OBJECTIVE: To study the mediating effects of L-selectin on the adherence of rat PMNs to the endothelia activated by burn sera. METHODS: Pulmonary microvascular endothelial cells (PMEC) of rat were cultured by tissue block method. Rat PMNs were harvested by continuous perfusion. The samples were collected for the detection of the adhering rate of PMEC to the PMNs after being cultured with L selectin monoclonal antibody (mAb), L-selectin ligand and sulfatide, respectively. RESULTS: The adhering rate of PMNs to PMEC was obviously lowered by the adding of L-selectin mAb and sulfatide. CONCLUSION: The adherence of PMNs to activated PMEC could be mediated by L-selectin. PMID- 11876950 TI - [An experimental study of the influence of burn on the expressions of IFN-gamma and IL-4 in T lymphocytes and on the expression of IL-12 in macrophages in mice]. AB - OBJECTIVE: To investigate the influence of burn on the expressions of IFN-gamma and IL-4 in T lymphocytes and on the expression of IL-12 in macrophages in mice. METHODS: The mice inflicted by 12% TBSA burn on the back were employed as the model. The peritoneal macrophages (PMphis) were isolated and counted under light microscope (LM). The gene expression level of IL-12 P35 and P40 subunits was determined with RT-PCR. The ratio of spleen to body and splenic histology were also determined. Splenic T lymphocytes were isolated and the gene expression levels of IFN-gamma and IL-4 in the T lymphocytes were examined by RT-PCR. RESULTS: (1) PMphis decreased significantly on 3 postburn day (PBD) and recovered quickly thereafter. (2) The gene expressions of IL-12 P35 and P40 subunits in PMphis were enhanced obviously. (3) Splenic index increased evidently and the karyokinesis phase increased in splenic lymphocytes. (4) The expressions of IFN gamma and IL-4 in splenic T lymphocytes increased postburn. CONCLUSION: The expressions of IFN-gamma and IL-4 in splenic T lymphocytes and the expressions of IL-12 in PMphis could be enhanced by burn injury. PMID- 11876951 TI - [Establishment and biological characteristics of a novel erythroleukemia cell line (HIE1)]. AB - OBJECTIVE: To establish a novel leukemia cell line and characterize its biological characteristics. METHOD: The cell line was established by liquid cell culture. The genetic marker was analyzed by R-banding and reverse transcriptase polymerase chain reaction (RT-PCR), cell morphology by microscopy, electron microscopy and histochemical staining, cell surface antigen by monoclonal antibody, hemoglobin by hyperomethemoglobin measurement and electrophoresis, erythroid differentiation by benzidine-staining, and monocyte-macrophage differentiation by cell morphology and phagocytosis. RESULTS: A novel erythroleukemia cell line (HIE1), with original cell genetic marker (Ph chromosome, bcr/abl fusion gene rearrangement), was established from a CML patient in blast crisis, and has been passaged for over 60 generations. Myelomonocyte marker and hemoglycoprotein A were found on the cell surface. HIE1 cells contained hemoglobin, the same HbA and HbA(2) bands as in normal individuals were displayed by Hb electrophoresis. The benzidine positive HIE1 cells were induced after exposure to 3.6 x 10(-4) mmol/L Ara-C. When HIE1 cells were treated with 100 ng/ml PMA for 3 days, one third of the cells became spindle in shape, and 6.5% of the cells exert phagocytosis. The cells were classified into two types with Wright-staining: one showing light blue cytoplasm and a few of cells with basophilic granules, the other showing dark blue cytoplasm with vacuoles and pseudopods without granules. In addition, POX, SB, CE stains were negative, and AE, PAS, ACP stains positive. Colony formation of the cells was 37%, the cell doubling time was 22 - 24 hrs, and EB virus detection was positive. CONCLUSION: A novel erythroleukemia cell line with bcr/abl fusion gene and characteristics of myelomonocytic and erythroid cells was established. PMID- 11876952 TI - [The clinical significance of lung resistance protein (LRP) gene expression in patients with acute leukemia]. AB - OBJECTIVE: To investigate the relationship between the expression of lung resistance protein (LRP) gene and drug resistance in patients with acute leukemias (AL). METHODS: Semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR)was used to examine the expression of LRP gene in AL patients and 15 normal subjects. Beta(2) microglobulin (beta(2)MG) was used as internal reference. LRP/beta(2)MG ratio >or= 0.3 was defined as LRP positive. RESULTS: The positivity percentage of LRP gene expression in newly diagnosed group was 32.4%. The first complete remission rate was 84.0% and 33.0% in LRP negative and LRP positive patients, respectively. The difference was significant (P < 0.005). The expression level of LRP mRNA and the positivity percentage of LRP in relapsed/refractory group were significantly higher than that in newly diagnosed group (P < 0.01). The expression level of LRP gene in normal subjects and long term survival groups was very low and correlated with FAB subtypes. The mdr-1 gene was examined simultaneously in 61 AL patients. No significant correlation was found between the expression of LRP and mdr-1 gene (P > 0.5). Coexpression of LRP and mdr-1 genes in the same AL patient might result in the worst prognosis. CONCLUSION: High expression of LRP gene leads to clinical drug resistance and is an unfavorable factor to AL patients of prognosis. PMID- 11876954 TI - [Current status of complete remission rate of acute leukemia in Shanghai]. AB - OBJECTIVE: To investigate the treatment outcome in newly diagnosed acute leukemia patients. METHODS: Two thousand five hundred and eighteen patients with acute leukemia registered in Shanghai were retrospectively analyzed. RESULTS: and CONCLUSIONS: The overall complete remission (CR) rate of acute leukemia was 60.4%: 51% for acute myeloid leukemia (AML) and 76.7% for acute lymphoid leukemia (ALL). CR rate of childhood ALL was 87.1%, while of elderly AML only 26%. Between 1984 and 1994, CR rates of AML and of ALL were not increased except for that of AML-M(3). Induction regimen was associated with CR rate. All-transretinoic acid was effective for AML-M(3). The VDLP and VDP regimens were more effective than that of VP regimen for ALL patients. There was no significant difference in CR rate between DA and HA regimens for treatment of ALL patients. PMID- 11876953 TI - [Comparative study of clinical features of childhood and adult acute lymphoblastic leukemia]. AB - OBJECTIVE: To analyze the outcome, the immunological and cytogenetic characteristics and the frequency of primary multi-drug resistance of childhood acute lymphoblastic leukemia (ALL) in our hospital during the past two years. METHOD: The complete remission (CR) rate of 154 newly diagnosed ALL patients were analysed and the biological features of the leukemic cells were explored by immunochemistry and cytogenetics. RESULTS: The CR rate in the ALL children is 94.1% which is significantly higher than that of adult ALL patients (67.8%). Immunophenotype analysis showed that 18.5% of the children expressed myeloid antigens besides lymphoid antigens and 4.8% of them were Ph chromosome positive. Both the figures were lower than that of adult patients. CONCLUSION: Childhood ALL has distinct morphological, immunological and cytogenetic (MIC) features, which might contribute to the good prognosis of the patients. PMID- 11876955 TI - [B7 and DC vaccines induced anti-tumor immunity against murine T-lymphocyte leukemia L615]. AB - OBJECTIVE: To study the effect of B7 and DC vaccines in inducing anti-tumor immunity in murine T-lymphocyte leukemia L615. METHODS: In vivo, murine mortality and survival were observed to compare the difference between B7 and DC vaccine in inducing immunoprotection against the subsequent challenge of live L615 cells;in vitro, specific cytotoxic assay and MLR were performed to test the specific cytotoxicity and proliferative activity of effective T cells. RESULTS: Both vaccines could efficiently improve T cell mediated anti-leukemic immunity in syngenic hosts, and DC vaccine was revealed to be more efficient. In vitro observations showed that both vaccines could induce tumor specific cytotoxicity and proliferative activity of effective T cells. CONCLUSION: DC vaccine is promising in tumor immunotherapy owing to its safety, efficiency and convenience. PMID- 11876956 TI - [Acute promyelocytic leukemia cell differentiation induced by tanshinone II A and its molecular mechanism]. AB - OBJECTIVE: To investigate APL cell differentiation induced by tanshinone II (Tan II A) and its molecular mechanism. METHODS: In vitro incubation of NB4 cells with Tan II A at the concentration of 0.5 microg/ml for 5 days, the cell differentiation was observed by cytomorphology, and nitroblue tetrazolium (NBT) test. Cell cycle, membrane CD(33), CD(11b) antigens and gene expressions (c-myc, c-fos, p53 and bcl-2) were analysed by flow cytometry. RESULTS: (91.3 +/- 2.1)% of NB4 cells were induced into morphologically and functionally more differentiated cells including 0.26 of myelocytes and metamyelocytes, and 0.68 of band form and neutrophils. Cell growth curve showed that growth of NB4 cells were inhibited. NBT reduction was significantly increased. Expression of CD(33) decreased and CD(11b) increased. The degrees of cell differentiation and growth inhibition induced by Tan II A or ATRA were no difference. Flow cytometry analysis showed that Tan II A arrested NB4 cell in G(0)/G(1) phase, inhibited cellular DNA synthesis, down-regulated c-myc and bcl-2 genes expression, and up regulated c-fos and p53 genes expression. CONCLUSION: Tan II A can induce differentiation and growth inhibition of NB4 cells. Its possible molecular mechanism might relate to modulation of gene expressions associated proliferation and differentiation, and to inhibition of DNA synthesis. PMID- 11876957 TI - [Detection of minimal residual disease in childhood hematological malignancies and its clinical significance]. AB - OBJECTIVE: Exploring the detection of minimal residual malignant cells in bone marrow from children with hematological malignancies to predict the prognosis. METHODS: Seventy-five patients with acute lymphoblastic leukemia (ALL), stage IV non-Hodgkin's lymphoma or stage IV neuroblastoma were studied. Complete remission was maintained for over 3 months before the detection. Minimal residual disease was detected by polymerase chain reaction (PCR) for IgH and TcRgamma rearrangements in lymphoid tumors by reverse transcriptase-polymerase chain reaction (RT-PCR) for neuroblastoma patients. RESULTS: Thirty five patients were positive for minimal residual disease, and 21 of them (60%) relapsed 3 - 40 months later, while only 7 (17%) negative patients relapsed (chi(2) = 12.59, P < 0.01). CONCLUSION: Minimal residual disease detection in bone marrow by PCR might predict prognosis in some childhood hematological malignancies. PMID- 11876958 TI - [Study on the antitumor activity of CBMC activated by IL-12 alone or in combination with IL-2]. AB - OBJECTIVE: To investigate the efficacy of cord blood mononuclear cells (CBMC) activated by interleukin-12 (IL-12) alone or in combination with interleukin-2 (IL-2) in tumor cells. METHODS: The cytotoxicity of CBMC and peripheral blood mononuclear cells (PBMC) activated by IL-12 and/or IL-2 was measured with (3)H TdR release assay, and the morphological change of K562 cells incubated with activated CBMC was observed. RESULTS: (1) CBMC activated by 10 IU/ml IL-12 produced significant cytotoxic activities which were (27.23 +/- 4.92)% and (29.12 +/- 3.46)% against K562 and Raji cells, respectively; moreover, 10 IU/ml IL-12 could be synergized with the same dose of IL-2 in enhancing the cytotoxicities which were (47.60 +/- 4.60)% and (38.69 +/- 4.86)% against the two target cells, respectively. (2) Short-term incubation with IL-12 activated CBMC increased the cytotoxicity to K562 cells, and long-term incubation increased the cytotoxicity to Raji cells. (3) No NK lytic activity of CBMC could be detected before cytokines stimulation. After stimulated by 10 IU/ml IL-12, NK lytic activity of CBMC was similar to that of PBMC. (4) K562 cells demonstrated obvious apoptosis after cultured with activated CBMC. CONCLUSION: CBMC activated by IL-12 had evident antitumor activity, which was further increased when IL-12 in combination with IL-2. These results showed the potential value to improve the graft-versus leukemia (GVL) response after cord blood progenitor cell transplantation. PMID- 11876959 TI - [Distribution of minimal residual leukemia in Brown Norway rats express LacZ gene]. AB - OBJECTIVE: To investigate the distribution of minimal residual leukemia (MRL) in vivo. METHODS: The MRL model in BN rat was established by using LT12nl leukemia cell line carrying LacZ gene. The marker gene (LacZ) was detected by using reverse transcriptase polymerase chain reaction (RT-PCR) and nested PCR, X-gal staining, and cytomorphological and pathological examination. RESULTS: PCR assays showed positive result in humerus at day 4, femur at day 6, and spleen at day 9 after CTX treatment. The peak positive signal in humerus was stronger at day 9. X gal staining showed negative in spleen at day 9. PCR showed negative results in peripheral blood and liver. CONCLUSION: PCR can be used as an effective tool to investigate the distribution pattern in experimental MRL. PMID- 11876960 TI - [Seven years' summary report on the treatment of acute promyelocytic leukemia with arsenic trioxide--an analysis of 242 cases]. AB - OBJECTIVE: To observe the effectiveness of arsenic trioxide (As(2)O(3)) in the treatment of acute promyelocytic leukemia (APL). METHODS: Data of clinical observation, seven year follow-up survey were analysed with American SAS system software, concentration of arsenic in cerebrospinal fluid (CSF) was determined by atomic absorbing spectrum. RESULTS: The complete remission (CR) and effectiveness rates were 87.9% and 94.4% for the previously untreated group; 60.0% and 65.0% for the relapsed in As(2)O(3) maintenance treatment group; 69.5% and 84.8% for the relapsed in other than As(2)O(3) maintenance treatment group; and 48.7% and 94.1% for the refractory group, respectively. In the previously untreated group, the relapse rate at 3 years after CR was 26.7%. The probability of survival in 136 follow-up survey patients was 92.0% and 76.7% at 5 years and 7 years, respectively. There were mild side effects. Central nervous system leukemia (CNSL) occurred in 138 out of 412 cases in 6 years, including some refractory leukemia, extramedullary residual leukemia and extramedullary relapse. Prophylactic intrathecal injection could reduce the incidence rate of CNSL. Determination of arsenic concentration indicated that it was difficult for arsenic to penetrate the blood-brain barrier. CONCLUSION: The CR rate and long term survival are high and the relapse rate is low in APL patients treated with As(2)O(3). There is no cross-resistance between ATRA and As(2)O(3) and other chemotherapeutic agents. As(2)O(3)-resistance occurs scarcely. It can also be used in maintenance treatment. PMID- 11876961 TI - [Chromosome t (4; 11) acute lymphoblastic leukemia: an analysis of 10 cases]. AB - OBJECTIVE: To characterize morphologically, immunophenotypically, cytogenetically and clinically the acute lymphoblastic leukemia (ALL) with t (4; 11) (q21; q23). METHODS: Ten cases of t (4; 11) ALL were reported. Of them, eight patients were de novo and two relapsed. The patients were treated with combination chemotherapy of DOPL or VP regimen. Immunophenotypic analysis was performed by flow cytometry in seven cases. Cytogenetic analysis was performed using bone marrow cells prepared directly and/or after 24h culture (case 7 using peripheral blood). RHG banding was used for karyotypic analysis. RESULTS: Ten (3.5%) of 285 ALL patients were found to be t (4; 11) ALL. In these 10 patients, increased WBC (> 100 x 10(9)/L) was found in 6 cases. Liver, spleen and/or lymph nodes were involved in all. t (4; 11) was detected as a single abnormality in 5 karyotypes whereas the other 5 showed additional aberration besides t (4; 11). Six patients were CD(19) (+), five were CD(22) (+) and one was HLA-DR (+), CD(5) (+), supporting that most cases of t (4; 11) ALL were B cell origin. Median survival time was 5.5 months in the present series. CONCLUSION: t (4; 11) ALL was a subtype with unique clinical and cytogenetic features, and had a poor prognosis. PMID- 11876962 TI - [Treatment of acute lymphoblastic leukemia by autologous stem cell transplantation: an analysis of 30 cases]. AB - OBJECTIVE: To evaluate the clinical outcome of autologous stem cell transplantation (ASCT) in acute lymphoblastic leukemia (ALL) and the affected factors. METHODS: Data of 30 ALL patients received ASCT in our hospital between July 1987 and December 1997 were retrospectively analyzed. Twenty-four of them were in the first complete remission (CR(1)) and six in the second complete remission (CR(2)) or early relapse (ER). Conditioning regimens were CTX 120 mg/kg + single total body irradiation 9 - 10 Gy (sTBI) or Bu 16 mg/kg of Mel 140 - 180 mg/m(2) + Ara-c 2 - 4 g/m(2). RESULTS: All patients reconstituted hematopoiesis. The median follow-up duration was 504 (18-3043) days. Transplant-related mortality was 10%. The probabilities of 3 year disease-free survival (DFS) for ALL in CR(1) and CR(2) were 67.7% +/- 10.3% and 16.7% +/- 15.2%, respectively (P = 0.00547); the 3 year DFS was significantly better with posttransplant treatment than without it (92.3% +/- 7.4% vs 50.0% +/- 17.7%, P = 0.0130). CONCLUSION: Acute lymphoblastic leukemia patients without HLA-matched related donor are recommended for ASCT in CR(1). To reduce relapse and improve the outcome, adoptive immunotherapy or maintenance chemotherapy should be given after ASCT. PMID- 11876963 TI - [Study on the differentiation and apoptosis of promyelocytic leukemia cells induced by tributyrin]. AB - OBJECTIVE: To investigate the differentiation of NB4 and MR2 cells induced by tributyrin (TB) in combination with different doses of all-trans retinoic acid (ATRA) and to see if TB can induce apoptosis of NB4 and MR2 cells. METHODS: Cell differentiation was analyzed by NBT reduction and CD(11b), CD(14) and CD(33), apoptosis by morphology, DNA electrophoresis, flow cytometry (FCM) and TUNEL in situ hybridization, bcl-2 expression by reverse transcriptase polymerase chain reaction. RESULTS: 0.2 mmol/L TB in combination with different doses of ATRA could potentiate the differentiation of NB4 cells induced by ATRA alone. No MR2 cell could be markedly induced by ATRA alone or in combination with TB. Treated with 1 mmol/L TB for 24 hours NB4 and MR2 cells exhibited a morphological characteristic of apoptosis and typical DNA ladder on gel electrophoresis. FCM analysis showed that TB could interfere with cell cycle in NB4 and MR2 cells, with a similar ratio of sub-G(1) in both cell lines. TUNEL in situ hybridization confirmed these results. Further study showed that TB could gradually down regulated bcl-2 expression. CONCLUSION: TB exerts synergetic effect on ATRA induced differentiation and induces apoptosis in promyelocytic leukemic cells. PMID- 11876964 TI - [Study of graft-versus -leukemia activity of gamma-ray irradiated haplomismatched donor lymphocytes infusion]. AB - OBJECTIVE: To explore graft-versus-leukemia activity of 7.5 Gy gamma-ray irradiated haplomismatched donor lymphocytes infusion (DLI). METHODS: L615 cells loaded 615 (H2(k))/BALB/c (H-2(d)) F(1) mice were infused with irradiated haplo mismatched donor lymphocytes at 3, 5, 7 days after CTX (200 mg/kg) treatment which was given at the 3rd day after L615 cells inoculation. RESULTS: T-cells irradiated with 7.5 Gy gamma-ray lost their proliferative capacity but preserved their cytotoxic activity against allogeneic targets. Survival of these recipient mice was longer than that of control (t = 10.03, P < 0.001) with no severe graft versus-host disease (GVHD). CONCLUSION: 7.5 Gy gamma irradiation attenuated the GVHD inducing potential of allogeneic lymphocytes while preserving their graft versus-leukemia activity. It provides a simple modality to postpone and prevent leukemia relapse. PMID- 11876965 TI - [Acute myeloid leukemia M2and t (8; 19) (q22; q13)]. AB - OBJECTIVE: Report for the first time of two cases of acute myeloid leukemia (AML) M(2) with t (8; 19) (q22; q13). METHODS: Chromosome specimens were prepared by short-term culture of bone marrow cells and karyotype analyses were carried out using R-and G-banding techniques. Immunophenotyping of the blast cells was analyzed by flow cytometry with a panel of monoclonal antibodies. AML1/ETO fusion gene was tested by "nested" reverse transcriptase-polymerase chain reaction (RT PCR). RESULTS: Karyotype analyses showed 46, XX, t (8; 19) (q22; q13) [28]/46, XX [2] in case 1 and t (8; 19) (q22; q13), del (9) (q12q22) [23]/46, XY [2] in case 2. In case 2, the blast cells expressed CD(13) (38.8%), CD(33) (31.8%), CD(34) (80.9%), and CD(19) (63.9%) and RT-PCR assay revealed no AML1/ETO fusion gene transcript. CONCLUSION: t (8; 19) (q22; q13) is a variant form of t (8; 21) (q22; q22). Its molecular entity remains to be elucidate. PMID- 11876966 TI - [Current status of hematopoietic stem cell transplantation in China: an analysis of results registered at China Bone Marrow Transplantation Registry in 1998]. AB - OBJECTIVE: To explore the application of stem cell transplantation (SCT) and the strategy for improving the SCT outcome. METHODS: Summarizing the data registered at China Bone Marrow Transplantation Registry (CBMTR) in 1998 to estimate the probabilities of disease free survival (DFS). RESULTS: There were 68 SCT teams registered. Thirty of them performed both allogeneic -SCT (allo-SCT) and autologous-SCT (ASCT). Up to Aug 31, 1998, a total of 1,840 SCT cases were registered, 38.6% of them were allo-SCT, 59.0% were ASCT, and 9 cases were syngeneic SCT. The probabilities of 3 year DFS for acute myeloid leukemia (AML) and acute lymphoid leukemia (ALL) in first complete remission (CR(1)) were 70.0% and 48.2%, respectively. For chronic myeloid leukemia (CML) in first chronic phase (CP(1)), 3 year DFS was 64.2%. The probability of 3 year DFS of ASCT for the AML-CR(1) patients was similar to that of the AML > CR(1) patients, but after ASCT, the ALL > CR(1) patients had much less probability of 3 year DFS than that of the CR(1) patients. There were 5 cases of volunteer marrow transplantation (VMT) registered. Two of them are alive, two died of relapse, and one died of infection. CONCLUSION: SCT including VMT should be performed while patients were in an ideal state of remission. PMID- 11876967 TI - [Factor XIIa-inhibited diluted thromboplastin time can reflect coagulation process]. AB - OBJECTIVE: To establish a screening test that can reflect two stages of coagulation process. METHODS: With FXII a being blocked, the effects of various dilution of thromboplastin on clotting time were observed. FX a activity was determined by chromogenic assay. RESULTS: (1) At high concentration of thromboplastin, FXII a-inhibited diluted thromboplastin time (FXII ai DTT) of pooled normal plasma and FXI deficient plasma was very similar, but at low concentration, FXII ai DTT was in order of FVIII and FIX deficient plasma > FXI deficient plasma > pooled normal plasma. (2) FXI consumption by immunologic method induced FXII ai DTT of pooled normal plasma prolonged, and addition of FXI to FXI deficient plasma shortened FXII ai DTT. (3) Alpha-thrombin blocked by hirudin at different time (10 - 80 seconds) resulted in decreased FXa generation, and the earlier the block, the more the decrease. Under the same condition, the amount of FXa generation was in order of platelet-rich plasma > platelet-poor plasma > FXI deficient plasma. CONCLUSION: (1) Our data support the two-stage hypothesis and confirm the important role of FXI in the amplification stage. (2) FXII ai DTT as a screening test for coagulation process may be practicable. PMID- 11876968 TI - [Antiphospholipid thrombosis syndrome and acquired activated protein C resistance]. AB - OBJECTIVE: To explore the correlation between antiphospholipid antibodies (APA) activated protein C resistance (APCR) and antiphospholipid thrombosis (APL-T) syndrome and investigate the mechanism of thrombosis in APL-T syndrome. METHODS: ELISA, APTT-LA and APTT+/-APC methods were used to detect anticardiolipin antibodies (ACA), lupus anticoagulants (LA) and APCR in 20 APL-T syndrome patients respectively. RESULTS: Twenty patients were diagnosed as APL-T. The patients were classified in three ways. According to etiology, 14 of 100 systemic lupus erythematosus (SLE) patients were diagnosed as "secondary" APL-T, and 6 of 16 "unexplained" thrombosis and spontaneous abortion as "Primary" APL-T syndrome. According to antibodies, 12 of 20 patients with positive LA were diagnosed as lupus anticoagulant thrombosis syndrome (LA-T), 12 of 20 patients with positive ACA as anticardiolipin antibodies thrombosis syndrome (ACA-T). According to clinical presentation: 7 of 20 patients with deep vein thrombosis were classified as type I APL-T, 3 of 20 patients with cerebrovascular thrombosis as type II, 1 of 20 patients with vein cerebrovascular thrombosis and spontaneous abortion as type IV (mixing), and 9 of 20 patients with spontaneous abortion as type V (fetal wastage type). The ACAs and LA are strongly associated with APL-T. There is close relationship between APCR and APL-T. CONCLUSION: The acquired APCR might be an important reason for thrombosis in APL-T caused by APA and LA. PMID- 11876969 TI - [The difference of thrombin-induced TF/uPAR mRNAs expressions in cultured endothelial cells and U937 cell line]. AB - OBJECTIVE: To compare the effects of thrombin on the expression of the tissue factor (TF) and urokinase-type plasminogen activator receptor (uPAR) mRNA in the cultured vascular endothelial cells (VEC) and U937 cell line. METHODS: Reverse transcriptase-polymerase chain reaction (RT-PCR) was used to detect the mRNA levels of TF and uPAR genes in the cultured cells under stimulation of thrombin, inactivated thrombin (DFP-T), or at the presence of the inducers and anti thrombin receptor antibody (ATR-Ab). RESULTS: In U937 cells, the TF and uPAR mRNA expression induced by thrombin could be fully quenched with the ATR-Ab, but in VEC, even at the presence of ATR-Ab, there still was a little stimulative effect of thrombin revealed. DFP-T had no effect on the expression of TF and uPAR mRNA in U937 cells, but could elicit a faint reaction in VEC. CONCLUSION: The induction effects on TF and uPAR mRNA expression in both of the cultured cells by thrombin were mainly dependent on the TR activation, which may not be the unique pathway for thrombin to induce TF and uPAR mRNA expression in VEC. PMID- 11876970 TI - [Study of platelet membrane glycoproteins and its fibrinogen binding reaction in patients with cerebral infarction and diabetes mellitus]. AB - OBJECTIVE: To investigate the change of platelet membrane glycoproteins and its fibrinogen binding reaction in patients with cerebral infarction and type II diabetes mellitus (DM), and assess their value in the study of thrombosis and pro thrombosis state. METHODS: By flow cytometry, the platelet glycoprotein (GP) I b, GP II b, GP III a, GP II b- III a complex and P-selectin expression as well as fibrinogen binding reaction were analyzed in 85 patients with type II DM and 28 with cerebral infarction, and compared with that in 30 normal subjects. RESULTS: The platelet membrane GP II b- III a complex, P-selectin expression and fibrinogen binding reaction were higher, while the platelet surface GP I b was lower in the patients with cerebral infarction and type II DM with vascular disease than those in normal group. The GP II b and GP III a were not significantly changed. There was no difference of these parameters between the type II DM without vascular disease patients and normal group. CONCLUSION: The platelet fibrinogen binding reaction, P-selectin and GP II b- III a complex expression were increased while the GP I b decreased in patients with cerebral infarction and type II DM with vascular disease. PMID- 11876971 TI - [The inhibition of prourokinase gene transfer on deposition of platelets on rabbit carotid artery intima]. AB - OBJECTIVE: To observe the expression of pro-urokinase (proUK) gene in rabbit carotid artery transfected with replication-deficient adenovirus vector containing proUK gene (Ad/prouk) and the deposition of platelet on the same injured vessel. METHODS: Ad/proUK (Ad/proUK, 3 x 10(10) pfu/ml) was injected into the right carotid artery locally. Wild type adenovirus (Ad) was locally injected into the left carotid artery as self-control. The expression of proUK gene was investigated by immunohistochemistry assay. After injury of the gene-transfected vessel by electric stimulation, the deposition of (111) In-labeled platelet was quantitatively observed. The thrombosis was observed with HE-stained vessel section. RESULTS: There were lots of proUK granules in the endothelium of Ad/proUK gene-transfected vessel. The differences in (111) In-platelets deposition per gram dry weight vessel segment were significant between Ad/proUK and Ad transfected control vessels [(4.60 +/- 0.93) x 10(7)/g vs control (27.95 +/- 4.93) x 10(7)/g, P < 0.01)]. The HE-stained vessel section showed that there were only small thrombi in proUK gene-transfected vessel but massive thrombi almost blocked up the whole vessel in control vessel. CONCLUSION: The proUK gene transfected vessel can obviously inhibit (111) In-labeled platelet deposition on injured vessel and thus thrombosis. PMID- 11876972 TI - [Relationship between human herpesvirus 6 infection and idiopathic thrombocytopenic purpura]. AB - OBJECTIVE: To investigate the pathogenic role of human herpesvirus 6 (HHV-6) in idiopathic thrombocytopenic purpura (ITP). METHODS: HHV-6 DNA was examined by polymerase chain reaction (PCR) in bone marrow mononuclear cells (BMMNC) of 105 ITP patients. Human cytomegalovirus (HCMV) and parvovirus B(19) DNA were also examined in some cases. Platelet-associated antibodies (PAIg) were measured by competitive ELISA in 66 ITP patients. Serum IgG titer to HHV-6 was observed by indirect immunofluorescence assay in 19 ITP patients. RESULTS: (1) HHV-6 DNA positivity was 41.0% for ITP patients, significantly higher than that for control group (P < 0.05). HHV-6 DNA positivity for adult ITP patients, especially adult chronic cases was significantly higher than that for childhood patients (P < 0.025). Positivities for parvovirus B(19) and HCMV DNA were 24.1% and 9.7%, respectively. (2) HHV-6 DNA positivity for patients with abnormal level of PAIgG was significantly higher than that for patients with normal level of PAIgG (P < 0.05). (3) Patients coinfected with HHV-6 and B(19) or HCMV had more severe symptoms or poorer prognosis. CONCLUSION: HHV-6 infection might be associated with excessive PAIgG. Coinfection with HHV-6, B(19) or HCMV may cause more severe symptoms in ITP patients. PMID- 11876974 TI - [Establishment and identification of B cell lines with PNH phenotype]. AB - OBJECTIVE: To establish B cell lines from patients with paroxysmal nocturnal haemoglobinuria (PNH) and normal controls. METHODS: B lymphoid cells prepared from PNH patients and normal controls were infected with EB virus. Phenotype of the immortal cells was analyzed by flow cytometry. The immunomagnetic beads were used to select CD(55)(-) and CD(59)(-) B lymphoid cells, which were cloned by limiting dilution. The clonality of the cloned cells was confirmed by testing the pattern of Ig rearrangements using polymerase chain reaction (PCR). RESULTS: From 2 PNH patients and 2 normal controls, CD(55)(-) and CD(59)(-) B lymphoblastoid cell lines were established which had PNH phenotype and normal phenotype, respectively. CONCLUSION: B lymphoid cell lines from PNH patients can be obtained by EB virus infection. PMID- 11876973 TI - [The significance of the adhesion of CD(34)(+) stem/progenitor cells to fibronectin in the pathogenesis of chronic myelogenous leukemia]. AB - OBJECTIVE: To investigate the significance of the adhesion of CD(34)(+) stem/progenitor cells to fibronectin in the pathogenesis of chronic myelogenous leukemia (CML). METHODS: (1) Integrin beta(1) chain (CD(29)) and alpha(4) chain (CD(49d)) expressions on CD(34)(+) cells were measured by flow cytometry in 30 untreated CML patients in chronic phase (CML-CP) and 10 healthy donors. (2) The adhesion function of immunomagnetic beads selected CD(34)(+) cells to fibronectin was detected by crystal violet staining in 5 untreated CML-CP patients and 5 healthy donors. (3) The effect of fibronectin on CFU-GM colony formation of bone marrow cells in 3 CML patients and 3 healthy donors was observed by limited dilution liquid microculture. RESULTS: (1) There was no significant difference in CD(29) and CD(49d) expressions on CD(34)(+) bone marrow cells between CML patients and healthy donors. (2) The adhesion of CD(34)(+) bone marrow cells to fibronectin decreased significantly in CML patients as compared with that in healthy donors (P < 0.01). (3) The CFU-GM colony formation was significantly inhibited by fibronectin in healthy donors (P < 0.01), but not in CML patients. CONCLUSION: The adhesion function abnormality of CML CD(34)(+) bone marrow cells to fibronectin may underlie the abnormal proliferation of CML progenitors. PMID- 11876975 TI - [Study of the expression and function of c-kit receptor on the bone marrow mononuclear cells of the patients with myelodysplastic syndromes]. AB - OBJECTIVE: To determine the expression and function of c-kit receptor on bone marrow mononuclear cells (BMMNC) of the patients with myelodysplastic syndromes. METHODS: c-kit protein (CD(117)) was detected by direct immunofluorescence assay, c-kit mRNA by reverse transcriptive polymerase chain reaction (RT-PCR) and the function of c-kit receptor by cell culture. RESULTS: c-kit protein expression in MDS was significantly higher than that in control [(8.58 +/- 5.28)% vs (3.04 +/- 1.49)%, P < 0.05]. c-kit protein expression in RA patients was significantly lower than that in RAEB/RAEB-t patients [(5.12 +/- 2.13)% vs (10.01 +/- 5.07)%, P < 0.05]. The rate of c-kit protein expression was (32.43 +/- 18.16)% in AML cases transformed from MDS (t-AML). c-kit mRNA expression in MDS was correlated with c kit protein expression. Addition of interleukin-3 (IL-3), erythropoietin (Epo) and stem cell factor (SCF) to the semi-solid culture of BMMNC showed significant stimulating effects on CFU-GM and BFU-E formation in normal controls, but did not in MDS patients. CONCLUSION: The protein and mRNA expressions of c-kit receptor on BMMNC in MDS are higher than those in normal control, and the function of this receptor in MDS is abnormal. PMID- 11876976 TI - [Detection of trisomy 8 with interphase fluorescence in situ hybridization in myelodysplastic syndromes]. AB - OBJECTIVE: To explore the value of interphase fluorescence in situ hybridization (FISH) in the detection of trisomy 8 in myelodysplastic syndromes (MDS). METHODS: Conventional cytogenetics (CC) and interphase FISH using SpectrumGreen labelled chromosome 8 centromere specific probe were simultaneously carried out to detect trisomy 8 in 69 MDS and 6 normal individuals. RESULTS: Two hundred interphase cells were counted and cells with three green hybridization spots > 3% was assigned. Eleven cases displayed trisomy 8 by CC and were confirmed in 10 by FISH. In 7 cases, the percentage of trisomy 8 cells was significantly lower by FISH than by CC. Seven cases displayed trisomy 8 by FISH in 58 cases who did not show trisomy 8 by CC. Of the 7 cases, two had 2 and 3 marker chromosomes respectively, 4 had normal karyotypes. CONCLUSIONS: Interphase FISH was a useful method for the detection of trisomy 8 in MDS, especially in patients with normal karyotype or marker chromosome. It was a important complement to CC. PMID- 11876977 TI - [Study on clinical and laboratory features of preleukemia patients]. AB - OBJECTIVE: To explore prospective diagnostic criteria for preleukemia. METHODS: A case control study of the discrepancies of clinical and laboratory features between patients with preleukemia and those with chronic aplastic anemia (CAA) or atypical paroxysmal nocturnal hemoglubinuria (a-PNH). RESULTS: There were eight variables of significance: (1) lymphocytoid micromegakaryocytes in marrow; (2) immature granulocytes in peripheral blood; (3) >or= 2% myeloblasts in marrow; (4) positive periodic acid schiff (PAS) staining of nucleated erythrocytes; (5) myeloid differentiation index >or= 1.8; (6) clonal karyotypic abnormalities; (7) negative sister chromatid differentiation; (8) > 4.0 cluster/colony ratio of granulocyte-macrophage colony-forming units (CFU-GM). The following criteria was assigned: A: To meet (1) and at least two of the other seven variables; B: To meet at least four of the eight variables. All of the patients with preleukemia met A or B and none of the patients with CAA or a-PNH did. CONCLUSION: Preleukemia is different from CAA or a-PNH. It has its own clinical and laboratory features, which may be useful for prospective diagnosis. PMID- 11876978 TI - [Linkage analysis of the G6PD gene mutations and its Nla III polymorphic site]. AB - OBJECTIVE: To investigate the molecular and anthropologic features of glucose-6 phosphate dehydrogenase (G6PD) deficiency and the linkage between the Nla III polymorphic site within G6PD gene and three common Chinese G6PD mutations. METHODS: By using denatured gradient gel electrophoresis (DGGE) and allele specific oligonucleotide (ASO) probe dot blot hybridization, Nla III polymorphic site at minus sign13 bp upstream exon 12 within G6PD gene were screened in 54 males with normal G6PD activity and 66 G6PD deficient males. RESULTS: Thirty-two cases with cDNA1376 (G-->T), 21 with cDNA1388 (G-->A) and 13 with cDNA95 (A-->G) were detected in the 66 G6PD deficient males. Among 54 normal controls, T at -13 bp upstream exon 12 within G6PD gene were detected in 11 cases (20.4%, without Nla III polymorphic sites) while C in 43 cases (79.6%, with Nla III polymorphic sites). All 66 G6PD deficient males were linked with C at -13 bp upstream exon 12 (with Nla III polymorphic sites). CONCLUSION: The G6PD mutations cDNA1376 (G- >T), cDNA1388 (G-->A) and cDNA95 (A-->G) were linked with Nla III polymorphism. This feature will play a role in studying the complicated manifestations and anthropology of G6PD deficiency. PMID- 11876979 TI - [Comparative study of three common G6PD gene mutations in Yao and Han People in Guangxi]. AB - OBJECTIVE: To study G6PD gene mutation diversity in Yao and Han people in Guangxi and compare the genotypic frequencies. METHODS: The three common G6PD gene mutations in Chinese were investigated in 34 G6PD deficient individuals from Yao and 37 from Han people by using an amplification refractory mutation system (ARMS). RESULTS: Among 34 cases of G6PD deficiency of Yao people, the genotypic frequencies of G1376T, G1388A and A95G were 41.2%, 26.5% and 14.7%, respectively. A case of C1311T was detected. As compared with that in Han people, the respective frequencies were 16.2%, 40.6% and 5.4%. CONCLUSION: The main G6PD mutations in Yao people were the same with Han people in Guangxi, but G1376T mutation was higher than that in Han people. The C1311T mutation in Yao people in Guangxi was reported for the first time. PMID- 11876980 TI - [Rapid detection of alpha-thalassemia of Southeast Asian deletion by polymerase chain reaction and its application to prenatal diagnosis]. AB - OBJECTIVE: To establish a rapid and simple polymerase chain reaction (PCR) method for detecting alpha-thalassemia of Southeast Asia deletion, and apply it to the prenatal diagnosis for high risk fetuses. METHODS: Two pairs of primers were designed: one pair bridging the breakpoints to identify the specific deletion, the other located in the common deletion region of --(SEA), -alpha(3.7) and alpha(4.2) gene to detect the normal chromosomes. In this system, the two amplifications ran in the same PCR tube under identical condition. RESULTS: A 740 bp fragment was amplified in chromosomes with --(SEA) determinant and a 1,052 bp fragment in normal chromosomes. For prenatal diagnosis, 3 of 8 at-risk cases were diagnosed as normal, 3 as heterozygotes, and 2 as homozygotes of --(SEA) deletion. CONCLUSION: This detection method is rapid and accurate and can be used as a routine method for carrier detection and prenatal diagnosis. PMID- 11876981 TI - [Molecular diagnosis in a Korean family with thalassemia intermedia due to co inheritance of triplicated alpha-globin genes (alphaalpha/alphaalphaalpha(anti 3.7)) and beta-thalassemia trait (IVS-II-1 G-->A)]. AB - OBJECTIVE: To analyze the molecular abnormalities of beta-thalassemia intermedia in a Korean family with thalassemia intermedia. METHODS: Polymerase chain reaction (PCR), Southern blot hybridization and double strand DNA cycle sequencing were used to analyse alpha, beta and gamma globin gene organization. RESULTS: In the Korean family the interaction between a triplicated alpha-globin locus and a heterozygous beta-thalassemia gave rise to a clinical phenotype of thalassemia. The molecular defect was a heterozygosity for a single beta thalassemia mutation (beta IVS-II-1 G-->A) and a triplicated alpha-globin gene (alphaalpha/alphaalphaalpha(anti 3.7)). CONCLUSION: Beta-thalassemia heterozygotes conjuncted with alpha-globin gene triplication was the major cause of the beta-thalassemia intermedia in this Korean family. PMID- 11876982 TI - [The expression of Fas antigen and plasma levels of sFas and sFasL in patients with aplastic anemia]. AB - OBJECTIVE: To investigate the role of Fas/FasL system in the pathogenesis of aplastic anemia (AA) and its correlation with clinical status. METHODS: Plasma levels of sFas and sFasL and the expression of Fas antigen on mononuclear cell (MNC) membrane in 32 AA patients and 24 normal controls were assayed by ELISA and FACS, respectively. RESULTS: The percentage of CD(34)(+)Fas(+) cells was significantly higher (P < 0.05) in AA patients than that in normal controls, and the percentage of CD(34)(+)Fas(+) cells in severe AA patients was much higher than that in chronic AA patients (P < 0.05). The percentage of CD(34)(+)Fas(+) cells was negatively correlated with the course of disease. The levels of sFas were significantly lower (P < 0.05) in AA patients than that in normal controls, and the sFas levels in severe AA were much lower than those in chronic AA (P < 0.05). The percentage of CD(34)(+)Fas(+) cells was negatively correlated with sFas level. CONCLUSION: Fas/FasL system aberration is involved in the pathogenesis of AA. PMID- 11876983 TI - [Effects of holothuria glycosaminoglycan on the expression of tissue factor and thrombomodulin in stimulated endothelial cells]. AB - OBJECTIVE: To study the mechanism of antithrombotic effects of holothuria glycosaminoglycan (GAG). METHODS: Endothelial cells from human umbilical vein which were pre-induced with 1 mg/L lipopolysaccharide, were treated by GAG (1 mg/L, 5 mg/L, and 10 mg/L, respectively) and 5 mg/L heparin as a control for 6 hours. Procoagulant activity (PCA), the expressions of tissue factor (TF) antigen and thrombomodulin (TM) antigen and their mRNA transcriptions were investigated. RESULTS: GAG could down-regulate the expression of TF antigen and mRNA, up regulate the expression of TM antigen and mRNA, down-regulate the PCA of stimulated endothelial cells. CONCLUSION: Down-regulating TF expression and up regulating TM expression of stimulated endothelial cells may be one of the mechanisms of GAG antithrombosis. PMID- 11876984 TI - [Exploration of gene expression profiles of CD(34)(+) hematopoietic stem/progenitor cells based on large scale sequencing]. AB - OBJECTIVE: To set up a large scale sequencing system and explore the gene expression profiles of CD(34)(+) hematopoietic stem/progenitor cells (HSPCs). METHODS: CD(34)(+) cells were isolated from umbilical cord blood and subjected to cDNA library construction. A primary profile of gene expression in HSPCs was emerged by EST sequencing and bioinformatics analyzing. RESULTS: Among 9,866 ESTs thus obtained, 7 476 meaningful ESTs were clustered into 2,060 unique sequence species (USSs), representing 1,054 known gene species and 1,006 unknown gene fragments. The 1054 known genes were divided into 8 categories: (1) hematopoiesis associated: 73, (2) chromatin structure and cell division/apoptosis: 91, (3) signal transduction and receptors: 111, (4) cell structure/mobility: 48, (5) cell/organism defense/homeostasis: 41; (6) Gene expression (transcription, translation and modification): 265, (7) metabolism: 192; and (8) unclassified: 233. CONCLUSION: A gene expression profile including 1,054 known genes and 1006 new gene fragments of HSPCs was primarily obtained, which may lay a basis for the further study on the molecular mechanism of hematopoiesis regulation and provide candidates for new gene cloning. PMID- 11876986 TI - [Anti-CD40 McAb induces CD40+ malignant B cell proliferation arrest and apoptosis]. AB - OBJECTIVE: To study the expression of CD(40) molecule and the biological effects mediated by CD(40) molecules on malignant B cells. METHODS: Agonistic anti-human CD(40) monoclonal antibody (5C11) was added to the cell culture system. Cell counting, PI staining, Annexin-V staining and flow cytometric analysis were used to study the biological effects of 5C11 on malignant B cell lines. RESULTS: 5C11 induced homotypic aggregation, proliferation arrest and apoptosis in the multiple myeloma cell line XG2 highly expressed CD(40). 5C11 induced B lymphoma cell line Daudi homotypic aggregation and proliferation arrest but no apoptosis. CONCLUSION: 5C11 can inhibit proliferation of malignant B cells by inducing apoptosis or arresting the cells at G(2)/M interphase. PMID- 11876985 TI - [Exploration of factors increasing transfer efficiency of retroviral mediated multidrug resistance gene (mdr1) into human CD(34)(+) cells]. AB - OBJECTIVE: To explore factors influencing transfer of retroviral mediated multidrug resistance gene (mdr1) into human CD(34)(+) cells. METHODS: Transduction efficiency in the presence of different combinations of cytokines and human bone marrow stromal cells plus cytokines were determined by flow cytometry (FCM). Drug resistance was evaluated by plating yields of cultured hematopoietic progenitor cells. The effect of Taxol at different concentrations on mdr1 gene transfer cells was determined by FCM. RESULT: Transduction efficiency in the presence of SCF + FL + IL-3 was higher than that in other combinations of cytokines (SCF + IL-6 + IL-3, SCF + IL-6 + IL-3 + Tpo, SCF + IL 3). Transduction efficiency (20.5%) in the presence of bone marrow stromal cells plus cytokines (SCF + FL + IL-3) was higher than that (15.2%) without stromal cells, and the number of drug-resistant colony-forming cells was more in the former than in the latter. The percentage of CD(34)(+) cell with the gene transduction at 10 ng/ml Taxol reached 38.5%. CONCLUSION: Human bone marrow stromal cells plus cytokines (SCF + FL + IL-3) is more effective in enhancing mdr1 gene transduction. Taxol at a certain concentration can enrich mdr1 gene transferred cells. PMID- 11876987 TI - [Mechanism of tissue factor expression on NB4 cells down-regulated by all-trans retinoic acid and arsenic trioxide]. AB - OBJECTIVE: To investigate molecular mechanism of tissue factor (TF) expression on acute promyelocytic leukemia cell line NB4 cells down-regulated by all-trans retinoic acid (ATRA) and arsenic trioxide (As(2)O(3)). METHODS: Cyclohexamide (CHX) inhibition test for de novo protein synthesis and actinomycin D (Act D) inhibition test for RNA synthesis were used to check the effect of ATRA on the TF expression. TF antigen of U937 cells transfected with pMSCV-PML-RARalpha treated with or without ATRA and As(2)O(3) was detected. RESULTS: CHX treatment completely suppressed the down-regulation effect of ATRA on the TF mRNA expression, Act D inhibition test showed that half-life of TF mRNA in treated NB4 cells was shortened to about 30 min from that of around 60 min in untreated NB4 cells. The TF antigen contents in U937 cells transfected with pMSCV-PML-RARalpha were significantly higher than that in transfected U937 cells with retrovirus vector. Both ATRA and As(2)O(3) could down-regulate the TF antigen level in U937 cells transfected with or without PML-RARalpha. CONCLUSION: The modulation of the TF mRNA expression in NB4 cells by ATRA might be indirect. TF mRNA destabilization was involved in the TF regulation process mediated by ATRA. Elevated TF antigen level in U937 cells transfected with pMSCV-PML-RARalpha may be related to the fusion protein PML-RARalpha. The down-regulation effect of ATRA and As(2)O(3) on the TF expression of U937 cells might not involve the fusion protein. PMID- 11876988 TI - [Proliferation inhibition and apoptosis induction of K562 cells by fatty acid synthase inhibitor--cerulenin]. AB - OBJECTIVE: To investigate the effect of fatty acid synthase (FAS) inhibitor- cerulenin on K562 leukemia cells and its mechanism. METHODS: Inhibition rate of cerulenin on K562 leukemia cells was assayed by MTT method, cell apoptosis by flow cytometry (FCM) and agarose gel electrophoresis. RESULTS: When treated with 10(-9) - 10(-5) mol/L of cerulenin for 24 h, the proliferation of K562 cells was obviously inhibited with dose related effect. At the same concentrations, the inhibition rates of human skin fibroblasts were all lower than 30%. When K562 cells were treated for 12 h with 50 microg/ml and 60 microg/ml of cerulenin, the apoptosis rate revealed by FCM was 42.30% and 38.8%, respectively, and DNA agarose gel electrophoresis showed the typical DNA ladder of apoptosis. CONCLUSION: Fatty acid synthase inhibitor--cerulenin inhibits proliferation of K562 cells but not of human fibroblasts. Cerulenin mediated cytotoxity is due to apoptosis induction. Fatty acid synthase might be a potential target for anti leukemia. PMID- 11876989 TI - [Mobilization of autologous peripheral blood stem cells by chemotherapy and recombinant granulocyte colony-stimulating factor (G-CSF) and granulocyte macrophage colony-stimulating factor (GM-CSF)]. AB - OBJECTIVE: To observe the efficacy of chemotherapy and recombinant granulocyte colony-stimulating factor (G-CSF, Glycosylated) and granulocyte-macrophage colony stimulating factor (GM-CSF) in autologous peripheral blood stem cells (APBSC) mobilization. METHODS: The mobilization regimen: CBP 350 mg/m(2) intravenously injected at day 1, Vp16 350 mg/m(2) intravenously injected from day 1 to day 3. G CSF and GM-CSF 5 microg x kg(-1) x d(-1) each, subcutaneously injected and DXM 5 mg/d intramuscularly injected, from the day of white blood cell (WBC) recovery to (2.4 - 6.4) x 10(9)/L from nadir to the day before the end of APBSC harvesting. APBSC harvesting started when WBC > 20.0 x 10(9)/L and ended when accumulated mononuclear cells (MNC) > 5 x 10(8)/kg. CFU-GM assay and CD(34)(+) cells counting of the APBSC were performed. RESULTS: Twenty cases underwent APBSC mobilization. APBSC harvest began at day 22.15 +/- 3.66 for two successive days. Accumulated MNC was (5.93 +/- 1.62) x 10(8)/kg, CD(34)(+) cells (23.10 +/- 11.53) x 10(6)/kg and CFU-GM (3.44 +/- 2.85) x 10(5)/kg. No severe toxicity was observed. Hematopoiesis was well reconstituted in 8 patients received single and in 1 patient received double APBSC transplantations. CONCLUSION: Chemotherapy combined with G-CSF + GM-CSF was a safe and highly effective method for APBSC mobilization. PMID- 11876990 TI - [Detection of hypermethylation of p15 gene in patients with acute leukemia by polymerase chain reaction]. AB - OBJECTIVE: To explore the role of hypermethylation of p15 gene in the pathogenesis of acute leukemias. METHODS: DNA from 36 cases of acute leukemia were investigated using restriction enzyme digestion combined with polymerase chain reaction (PCR) technique. RESULTS: Hypermethylations of p15 gene were found in 22 (61.1%) patients, including 17 cases of acute non-lymphoblastic leukemia (ANLL) and 5 of acute lymphoblastic leukemia (ALL). ANLL patients exhibited p15 gene hypermethylations had higher WBC in peripheral blood (P < 0.05) and higher percentages of leukemia cells in bone marrow (P < 0.01). CONCLUSION: Inactivation of p15 gene due to hypermethylation may be involved in the pathogenesis of some acute leukemias. PMID- 11876991 TI - [The rearrangements of immunoglobulin heavy chain and T cell receptor delta genes in patients with acute nonlymphocytic leukemia]. AB - OBJECTIVE: To explore the clinical significance of rearrangements of immunoglobulin heavy chain (IgH) and T cell receptor (TCR) genes in patients with acute nonlymphocytic leukemias. METHODS: Polymerase chain reaction (PCR) was used to study IgH and TCRdelta genes rearrangements in 30 untreated acute nonlymphocytic leukemia patients. RESULTS: IgH gene rearrangements were found in 9 patients, and TCRdelta gene rearrangements in 5 patients. CONCLUSION: The lineage infidelity does occur in acute nonlymphocytic leukemia, showing not only a high frequency of IgH gene rearrangement, but also of TCRdelta gene rearrangement. PMID- 11876992 TI - [Construction and transient expression of an Ala 737-->Glu mutant of vWF]. AB - OBJECTIVE: To study the functional change of an Ala737-->Glu substitution mutant of vWF and the molecular pathological mechanism of this mutant in the type 2A vWD. METHODS: The expression plasmid pSVvWF containing full-length cDNA of vWF was used to site direct mutagenesis by polymerase chain reaction (PCR) and transitorily expressed in the COS-7 cells, vWF: Ag in the supernatant and the cell lysate between wild type and pSVAla737Glu vWF were measured. vWF polymers of the platelet of the patient with the mutation and response to DDAVP treatment of the patient were observed. RESULTS: vWF: Ag level of pSVAla 737Glu vWF was 76.4% and 98.8% of the wild types in the supernatant and cell lysate, respectively. The polymer pattern of extracellular pSVAla737Glu vWF was indistinguishable from that of the wild type, containing all kinds of molecular weight. Increased vWF antigen levels was observed in the patients after DDAVP treatment. CONCLUSION: This mutant did not change the assembly and secretion of vWF. The mutant of vWF Ala737 ->Glu resulted in Group II type 2A vWD. PMID- 11876993 TI - [In vitro induction of autologous T cell killing by heat treated human chronic myelogenous leukemia cells]. AB - OBJECTIVE: To investigate the potential of autologous T cell killing of heat treated chronic myelogenous leukemia (CML) cells (autologous tumor killing, ATK). METHODS: (51)Cr release assay was used to measure the ATK activity of autologous T cells against CML cells treated with 42 degrees C for 30 minutes (heat) or 37 degrees C for 30 minutes (non-heat). The phenotypes of T cells and heat shock protein 70 (HSP70) expression of CML cells were measured by flow cytometry (FCM). T cells from the CML patients were stimulated and expanded by autologous mixed lymphocyte/tumor cell cultures (MLTC). RESULTS: ATK activity of autologous T cells to the non-heated and heated CML cells were found in 4 (19.05%) and 10 (47.62%) of the 21 cases, respectively. The ATK activity of interleukin-2 (IL-2) stimulated autologous T cells against heated CML cells was markedly higher than that of unstimulated autologous T cells against non-heated CML cells (P < 0.001). FCM analysis showed that no HSP70 was expressed on the CML cell membranes whether heated or non-heated, but intracellular HSP70 expressions were (83.42 +/- 5.65)% and (78.34 +/- 6.32)% pre- and post-heated, respectively. The phenotypes of T cells stimulated and expanded in MLTC were TCRgammadelta - CD(3)(+), mostly CD(8)(+), with some activation markers (CD(25) and HLA-DR) expression. The ATK activities of these T cells against the heated and non-heated CML cells and K562 cells were (51.25 +/- 4.26)%, (36.52 +/- 3.83)% and (2.92 +/- 1.19)%, respectively. CONCLUSIONS: ATK activity of autologous T cells against CML cells could be induced or enhanced by heat treatment of the CML cells particularly of T cells stimulated with IL-2. This ATK activity was not associated with gammadelta T cells or HSP70 expression of CML cells. PMID- 11876994 TI - [A preliminary study on placenta hematopoiesis]. AB - OBJECTIVE: To study the role of placenta on fetal hematopoiesis during embryo ontogeny, so as to find a new source of hematopoietic stem/progenitor cells (HSC/HPC) for clinical transplantation. METHODS: Cord blood was collected separately from cord vein and artery. CD(34)(+) cells were detected by FACS, CFU GM semisolid agar culture, and hematopoietic growth factors (HGFs) by ELISA. The results of HE stain and CD(34) McAb immunohistochemistry stain of placenta chorionic villi were investigated. RESULTS: The CFU-GM yields and the percentage of CD(34)(+) and CD(34)(+)/CD(38)(minus sign) cells in cord vein were higher than that in artery. The cord vein contained larger amount of hematopoietic stimulating factors and similar level of hematopoietic inhibitory factors as compared with that in artery. Blood island, reticular cells and scattered CD(34)(+) cells were found in the mesenchyme of placenta chorionic villi. CONCLUSION: The placenta might be an another hematopoietic organ in ontogeny. It could be applied to clinical hematopoietic cell transplantation. PMID- 11876995 TI - [Proliferative signal transmission mediated by EpoR in leukemic cell lines]. AB - OBJECTIVE: To investigate the expression of erythropoietin receptor (EpoR) in leukemic cell lines and to clarify the mechanism of proliferative signal transmission in the leukemic cell line KOCL-33 mediated by EpoR. METHODS: Biotinylated Epo and flow cytometry were used for EpoR expression, (3)H-TdR incorporation for cell proliferation, immunoprecipitation and Western blot analysis for the tyrosine phosphorylation of signal transmission proteins. RESULTS: (1) All the cell lines tested expressed EpoR except for T-lymphoid cell lines. The positivity rates ranged from 18% to 99% with an average of 52%, and was no difference among T, B and non-lymphoid cells. (2) The cell proliferation was significantly enhanced in response to Epo in 7 of 9 cell lines tested. The stimulating effects of Epo did not correlate with the densities of EpoR expressed on the cells. (3) The proliferation of KOCL-33 cells was significantly enhanced in response to Epo. Dimerization and tyrosine phosphorylation of EpoR and tyrosine phosphorylation of JAK2 occurred in 1 min and tyrosine phosphorylation of STAT5 in 5 minutes after Epo stimulation. CONCLUSION: (1) There was EpoR expression in leukemic cell lines. (2) The cell proliferation could be stimulated by Epo in some leukemic cell lines. The stimulating effect of Epo did not correlate with the densities of EpoR expressed on the cells. (3) Dimerization and tyrosine phosphorylation of EpoR occurred, and JAK2, STAT5 involved in the proliferative signal transmission in KOCL-33 cells mediated by EpoR. PMID- 11876997 TI - [Dynamic analysis of junctional sequences of T cell receptor (TCR) delta and gamma gene rearrangement in childhood with acute lymphoblastic leukemia]. AB - OBJECTIVE: To explore the junctional sequence difference of T cell receptor delta and gamma gene rearrangement in childhood acute lymphoblastic leukemia (ALL) at diagnosis, complete remission (CR) and relapse. METHODS: By using the T-vector molecular cloning and sequencing and polymerase chain reaction, the junctional sequences of TCRdeltaV-D and TCRgammaV-J were dynamically analyzed in 34 bone marrow samples of ALL. RESULTS: The junctional sequence of TCR delta, gamma gene showed significant difference and regularity before and after remission and during relapsed periods. The sequence of TCRdeltaV-D were analyzed in 24 samples from ALL. Among them, the intact Vdelta2 and 5'Ddelta3 sequences were observed in 10 samples at diagnosis, of which 7 samples had T-->C mutation in Ddelta3 nonamer sequence. The deletions of rearranged Vdelta2, 5'Ddelta3 and Ddelta3 haptamer sequences were found in 11 complete remission (CR) samples with ALL, but none had T-->C mutation in Ddelta3 nonamer sequence. The deletion rate of Vdelta2 or Ddelta3 sequences and the T-->C mutation in Ddelta3 nonamer sequence were extremely differed between samples at diagnosis and in remission (calculate exact probabilities P = 0.001). The junctional rearrangement sequence of 5'Ddelta3 sequences tended to remain intact in 3 relapsed samples. The findings of TCRgammaV-J sequences were similar to that of TCRdeltaV-D in 10 ALL patients. CONCLUSION: The difference of TCRdeltaV-D and TCRgammaV-J junctional sequences were related to the development, therapeutic effectiveness and outcome in ALL. PMID- 11876996 TI - [Taxol-induced apoptosis in Jurkat T cell lymphoma cell line and its molecular mechanisms]. AB - OBJECTIVE: To observe whether antimicrotubular drug taxol can induce apoptosis in Jurkat T cell lymphoma cell line and the role of bcl-2 gene family in this process. METHODS: Different concentrations of taxol were used to treat Jurkat cells. Cell morphology was observed under light and electron microscope. Flow cytometry and electrophoresis were used to analyze DNA contents and DNA fragments. bcl-2 gene family proteins and mRNAs were studied by immunohistochemistry and semi-quantitative RT-PCR technique. RESULTS: Taxol could inhibit Jurkat cell growth. Within a certain range of treating time and dose, cells were induced apoptosis with a time and dose related manner. The expressions of bax protein and mRNA were increased and bcl-x(s) mRNA became detectable after taxol treatment. CONCLUSION: Taxol can specifically induce Jurkat cell apoptosis. It might provide a theoretical basis for clinical treatment and a good model for studying apoptotic gene modulation. Bax and bcl-x(s) participate in the taxol induced apoptosis of Jurkat cells. PMID- 11876998 TI - [Caspase 3 in the cytokines induced acute leukemia cell apoptosis]. AB - OBJECTIVE: To explore the variation and significance of caspase 3 activity in the process of G-CSF and GM-CSF regulated NB4 cell apoptosis. METHODS: Cell morphology, flow cytometry and DNA fragmentation assay were used to demonstrate the apoptosis induced by G-CSF and GM-CSF. Caspase 3 activity was detected by spectrofluorometry, and its inhibition test by AC-DEVD-CHO. RESULTS: G-CSF could induce NB4 cell apoptosis, while GM-CSF could not. Caspase 3 activity was significantly higher in the apoptotic cells than in control cells. The G-CSF induced NB4 cell apoptosis was efficiently inhibited by AC-DEVD-CHO. CONCLUSIONS: G-CSF and GM-CSF exert different effects on the apoptosis of leukemic cells. G CSF induced apoptosis of leukemic cells through activation of caspase 3. PMID- 11876999 TI - [Altered subcellular distribution of daunorubicin in the non-P-glycoprotein mediated multidrug-resistant cell line HL-60/ADR]. AB - OBJECTIVE: To investigate DNR subcellular distribution in the non-P-glycoprotein mediated multidrug-resistant cell line HL-60/ADR and its relation to multidrug resistance. METHODS: DNR subcellular disposition was studied by confocal scanning laser microscopy, fluorescent methods, MTT and RT-PCR. The effects of verapamil, brefeldin A, chloroquine were also examined. RESULTS: In the drug-sensitive cell line HL-60 DNR fluorescence distributed evenly in the nucleus and cytoplasm, while in the resistant cell line DNR distributed in a punctate pattern in the cytoplasm and was reduced in the nucleus. Verapamil, brefeldin A, but not chloroquine could recover the intracellular distribution of DNR from punctate to even in the resistant cell line. CONCLUSION: Altered subcellular disposition of DNR in resistant cell line was involved in the mechanism of multidrug resistance. PMID- 11877000 TI - [Effects of IFN-alpha combined with IL-6 on cell growth and related genes expression and apoptosis of bone marrow cells from CGL patients]. AB - OBJECTIVE: To investigate the effects of interferon-alpha (IFN-alpha) and IFN alpha combined with interleukin-6 (IL-6) on cell growth and bcr/abl, bcl-2 and c myc genes expression in the bone marrow mononuclear cells (MNC) from chronic granulocytic leukemia (CGL) patients. METHODS: MNCs were cultured in liquid medium at the presence of IFN-alpha (200 U/ml) or IFN-alpha (200 U/ml) plus IL-6 (100 ng/ml). The viable cells were counted and the expression levels of beta actin, bcr/abl, bcl-2 and c-myc genes were quantitatively detected by reverse transcriptase-polymerase chain reaction (RT-PCR). RESULTS: The cell growth was markedly inhibited by IFN-alpha, but the extent of the inhibition was slightly decreased when IFN-alpha combined with IL-6. The expression levels of bcr/abl and bcl-2 gene were reduced by IFN-alpha or IFN-alpha plus IL-6. The expression of c myc gene was inhibited by IFN-alpha but promoted by IL-6. CONCLUSIONS: Both IFN alpha and IFN-alpha plus IL-6 can inhibit the expression of anti-apoptosis genes, and modulate the expression of c-myc. It is the possible mechanism of IFN-alpha therapy for CGL in chronic phase. PMID- 11877001 TI - [Antigen loading on dendritic cells affects the cell function in stimulating T cells]. AB - OBJECTIVE: To study the effect of antigen loading on dendritic cells (DC). METHODS: DCs collected from peripheral blood monocytes were loaded with a tumor antigen from XG-7 cell line. These DCs were then co-cultured with allogeneic T cells and were compared with those DCs without antigen exposure. RESULTS: Although DCs showed no change in their phenotypes after cultured with the antigen, they secreted more IL-12, and became more powerful in allogenic mixed lymphocyte reaction (MLR). Antigen-loaded DC stimulated more CD(4)(+) cells to proliferate than nonantigen-loaded DCs did. These CD(4)(+) cells did not kill XG 7 cells, but promoted CD(8)(+) cells' ability to inhibit the XG-7 proliferation. CONCLUSION: The proliferation of CD(4)(+) cells after cultured with DC may become an indicator for the function of antigen loaded DC and for the efficiency of DC immunotherapy. PMID- 11877002 TI - [Study on clinical and molecular biological characteristics of infant acute leukemia]. AB - OBJECTIVE: To study the clinical and molecular biological characteristics of infant acute leukemia (IAL). METHODS: R and/or G banding technique was used for analysis of karyotype. DNA blotting for HRX gene rearrangement, and polymerase chain reaction (PCR) and reverse transcriptase PCR (RT-PCR) for fusion gene detection. RESULTS: Twenty cases of IAL were detected. HRX gene rearrangement was found in 10 cases, including HRX/AF-4 fusion gene in 5, HRX/AF-9 fusion in 2, and HRX/ENL fusion in 1, HRX self-fusion mediated by alu-repeat homologous recombination and HRX/EEN fusion each in one (HRX/EEN is a novel fusion gene reported for the first time). CONCLUSION: High frequency of HRX gene rearrangement occurred in IAL, which is characterised by a massive leukemia cell burden and 11q23 translocation, forming fusion genes, especially HRX/AF-4 (about 50%). The results are of important significance in guiding clinical treatment and approaching the etiology of IAL. PMID- 11877003 TI - [Study on the clinical characteristics of biphenotypic acute leukemia]. AB - OBJECTIVE: To analyze the biological characteristics and the treatment outcome of adult biphenotypic acute leukemia. METHODS: Immunophenotypes were examined using indirect immunofluorescence method. Biphenotypic acute leukemia (BAL) was diagnosed according to EGIL scoring system. RESULTS: (1) The incidence of BAL in acute leukemia was 3.4%. Percentage for coexpression of B lymphoid and myeloid antigens was 68.4%, for T lymphoid and myeloid antigens 21.1%, for B, T lymphoid and myeloid antigens 10.5%. (2) CD(34) was expressed in 43.75% of the BAL cases. (3) Cytogenetic analysis revealed normal and abnormal karyotypes in 41.7% and 58.3% of the BAL cases, respectively. (4) Six of 19 patients achieved completed remission (CR), but the disease free survivals were all less than 6 months. Treatment outcomes were negatively related to the expression of CD(34) antigen and cytogenetic findings. The BAL patients were poorly responded to therapeutic regimens directed to AML. CONCLUSION: Coexpression of B/M antigens is the commonest subtype in BAL. BAL had a poor prognosis, especially treated with induction regimen directed to AML. PMID- 11877004 TI - [Protection of hematopoietic stem cells by MIP-1alpha and PF4 against the cytotoxicity of chemotherapeutic agents]. AB - OBJECTIVE: To study the protective effects of macrophage inflammatory protein 1alpha (MIP-1alpha) and platelet factor 4 (PF4), alone and in combination, on hematopoietic stem/progenitor cells against the cytotoxicity of chemotherapeutic drugs. METHODS: Bone marrow and cord blood mononuclear cells (BMMNC and CBMNC) and HL-60 cells were pretreated with MIP-1alpha, PF4, MIP-1alpha + PF4, and PBS respectively for 48 hours, and then incubated with DNR for an additional 24 hours. Cell viability, cell cycle, CD(34)(+) CD(38)(-) cells, colony forming units (CFU) and protein expression of p16, p27 gene were measured. RESULTS: (1) Cell viability, the number of CD(34)(+) CD(38)(-) cells and CFU yields of BMMNC and CBMNC in MIP-1alpha and PF4 groups were significantly higher than that in control groups (P < 0.05). (2) Cells in S + G(2) phase in MIP-1alpha and PF4 groups were significantly fewer than that in control groups (P < 0.05). (3) MIP 1alpha upregulated the expression of p16 gene of stem/progenitor cells. PF4 showed no effects on expression of both p16 and p27 genes. (4) Hematopoietic protection of MIP-1alpha was stronger than that of PF4. No cooperative effect could be seen in combination of the two agents. (5) Cell viability, cell cycle and expression of p16 and p27 gene of HL-60 cells were not affected by either MIP 1alpha or PF4. CONCLUSION: MIP-1alpha and PF4 can reversibly and selectively protect hematopoietic stem/progenitor cells against cytotoxicity of chemotherapeutic agents. MIP-1alpha can suppress cell proliferation by upregulating the expression of p16 gene and block the cell cycle at G(0) phase, resulting in the elevation of cell resistance to chemotherapeutic drugs. PMID- 11877005 TI - [The mechanisms of inhibitory effect of bufalin on human leukemia cells]. AB - OBJECTIVE: To study the mechanism of bufalin on human leukemia cell inhibition. METHODS: HL-60 cells were treated with bufalin at different concentrations. The growth inhibition was analysed by MTT assay, cell apoptosis by light microscopy, transmission electron microscopy, flow cytometry, TUNEL labeling method and agarose gel electrophoresis. RESULTS: (1) Treatment of HL-60 cells with bufalin remarkably inhibited the cell growth, the IC(50) value of bufalin for HL-60 cells was 0.025 micromol/L. (2) Apoptosis of HL-60 cells could be efficiently induced by bufalin at concentration of 0.010 micromol/L or higher. (3) Bufalin induced apoptosis of HL-60 cells in a dose- and time-dependent manner. (4) With G(1) phase cells decreasing, S phase cells increased, and then apoptotic cells increased with a diminution of S phase cells. (5) Bufalin-induced apoptosis of HL 60 cells was inhibited by ZnCl(2), an inhibitor of endonuclease, but not by cycloheximide, an inhibitor of protein synthesis. CONCLUSION: Bufalin could efficiently induce apoptosis of HL-60 cells, especially the S phase cells. PMID- 11877007 TI - [Regulation of telomerase activity in HL-60 and NB4 cells by arsenic trioxide]. AB - OBJECTIVE: To investigate the regulation of telomerase activity in HL-60 and NB4 cells exposed to arsenic trioxide (As(2)O(3)). METHODS: Cell morphology, intracellular DNA distribution, bcl-2 expression and telomerase activity were evaluated in HL-60 and NB4 cells exposed to As(2)O(3) or ATRA. RESULTS: The differentiation induction of HL-60 and NB4 cells by low concentration of As(2)O(3) was weaker than that by ATRA, but downregulation effect of As(2)O(3) on telomerase activity was more strong and quick. Higher concentration of As(2)O(3) induced apoptosis in HL-60 or NB4 cells accompanied by more rapid down regulation of telomerase activity and bcl-2 expression. Neither As(2)O(3) nor ATRA showed direct inhibition effect on telomerase activity. CONCLUSION: The downregulation of telomerase activity by low concentration of As(2)O(3) in HL-60 and NB4 cells is the result of overlap of differentiation and apoptosis. There are similar mechanisms in the regulation of telomerase activity and apoptosis in different leukemia cells. Bcl-2 may play an important role in these mechanisms. PMID- 11877006 TI - [The St14 (DXS 52) VNTR in a Guangdong Han population and detection of hemophilia A carriers]. AB - OBJECTIVE: To investigate the genetic polymorphism of the St14 (DXS 52) variable number tandem repeat (VNTR) in normal individuals in Guangdong, and to evaluate the efficacy of this marker for the gene diagnosis of hemophilia A. METHODS: 125 unrelated healthy individuals (male 21, female 104) and 4 hemophilia A families were detected using amplified-fragment-length polymorphism (Amp-FLP). RESULTS: 11 allelic fragments ranging from 700 to 1,810 bp in size and 7 different genotypes in males, 17 different genotypes in females were observed, respectively. The allele frequencies were 0.0044 to 0.4803. The polymorphism information contents (PIC) was 0.7335, and the heterozygosity was 0.432. Four families with hemophilia A were analyzed and 2 of them were informative for linkage analysis. In one family, 2 females were determined to be normal individuals, not carriers, one female carrier was detected in the other family. CONCLUSION: St14 (DXS 52) was a useful polymorphism marker for carrier detection of hemophilia A in southern Chinese population, and it was different from those in Caucasian. PMID- 11877008 TI - [Comparison of serological and DNA typings for HLA-AB of cord blood samples]. AB - OBJECTIVE: To evaluate the practical value of serological method for HLA-AB typing of cord blood samples. METHODS: Serological typing results of 1800 cord blood samples were analyzed and 36 of them were compared with that of PCR-SSP (polymerase chain reaction-sequence specific primers). Lymphocytes in cord blood samples were isolated by T-immunomegnetic beads or Ficoll. RESULTS: T lymphocytes isolated by fluorobeads have better activity than that in mononuclear cells isolated by Ficoll. In order to get the more active cord blood cells, samples stored in vitro over 24 hours must be isolated with fluorobeads. Its serological reaction is lower as compared with that of peripheral mononuclear cells. Except 28 samples which had too many dead cells or few cells, there were 310 (17.5%) unconfirmed antigens among 1772 cases by serological typing. The main causes were low specificity of typing reagents, cross reaction, weak serological reaction, high background of dead cells and extra reaction. Confirmed by PCR-SSP, the serological typing had a wrong result rate of 13.88%. CONCLUSION: Although serological typing is rapid and easy, it is necessary to use genetic typing as a complement to it. PMID- 11877009 TI - [Autologous bone marrow transplantation for patients with chronic myelogenous leukemia after in vitro purging of the graft with bcr/abl antisense oligodeoxynucleotides]. AB - OBJECTIVE: To investigate the efficacy of autologous bone marrow transplantation (ABMT) for patients with chronic myelogenous leukemia (CML) after in vitro purging of the graft with bcr/abl antisense oligodeoxynucleotides (AS-ODN). METHODS: Five CML patients, 2 in chronic phase (CP), 1 in accelerated phase (AP) and 2 in blast crisis (BC), all confirmed the presence of b3a2 bcr/abl mRNA by RT PCR (reverse transcriptase-polymerase chain reaction). Patients had received 2 courses of intensive chemotherapy for "in vivo purging" before bone marrow harvesting. The autologous bone marrow cells were concentrated by CS3000plus and then incubated with 18-mer phosphorothioate bcr/abl AS-ODN (40 - 60 microg/ml, for 48 - 60 h). Conditioning regimens were TBI + Cy (total body irradiation + cyclophosphamide) or MAC + CCNU. RESULTS: After intensive chemotherapy, ph(+) cells in bone marrow reduced to 34% (24% - 46%) and the numbers of bcr/abl mRNA (+) CFU-GM to 45.6% (33% - 58%). After bcr/abl AS-ODN in vitro purging, two patients were bcr/abl mRNA negative and three positive with the number of bcr/abl mRNA (+) CFU-GM markedly reduced. Delayed hematopoietic reconstitutions were observed in all patients after ABMT. During a more than 2 years follow-up period, three patients obtained major cytogenetic remission (MCR) and lasted for 9 - 12 months and the durations of CP after ABMT were longer than that before ABMT. One of this three in BC before ABMT being survived disease-freely for 37 months with bcr/abl mRNA (-) after ABMT. Another patient in BC had a short duration of MCR and relapsed at +7 months. One patient died from severe infection and bleeding at day +74 because of delayed reconstitution. CONCLUSION: ABMT with bcr/abl AS-ODN purged graft can result in quite long duration of MCR and prolonged CP in some patients with CML. PMID- 11877010 TI - [The toxicity of busulphan and cyclophosphamide as the preparative regimen for allogeneic peripheral blood stem cell transplantation]. AB - OBJECTIVE: To analyse the treatment-related complications of busulphan and cyclophosphamide (BU-CY) as the conditioning regimen for allogeneic peripheral blood stem cell transplantation (allo-PBSCT). METHODS: The clinical data of 40 leukemia patients undergoing allo-PBSCT between June 1997 and May 1999 in our BMT center were retrospectively analysed. RESULTS: Recovery of neutrophil and platelet was achieved at a median of day +13 (9 - 28) and day +12 (7 - 60) respectively. Acute GVHD occurred in 17 of 40 patients (42.5%) with grade II-IV in 10 patients (25%). Chronic GVHD developed in 21 out of 30 evaluable patients (70%). Mild to severe mucositis occurred in 30 patients (75%), and 4 of them had severe esophagitis with bleeding. Haemorrhagic cystitis developed in 8/40 (20%) patients, the median time of its onset was day +100 (+7 to +165). Six of 40 patients (15%) developed interstitial pneumonia (IP), 5 of them were due to cytomegalovirus infection, and the remaining one due to pneumocystis carinii infection. No hepatic veno-occlusive disease was observed and no seizure occurred. During the median follow-up of 480 (300 - 1000) days, 4 (10%) patients relapsed and 8 (20%) patients died of the transplant-related complications. The 3 year leukemia-free survival rate was 70%. CONCLUSION: BU (domestic busulfan)-CY regimen is relatively easy to administer and well tolerated, with low extramedullary toxicities. PMID- 11877011 TI - [Evidence for TCR Vbeta clonal expansion T cells in a patient with cGVHD after unrelated donor BMT]. AB - OBJECTIVE: To investigate the distribution and clonal expansion of TCR Vbeta subfamily T cells in patients with cGVHD after unrelated donor bone marrow transplantation (BMT). METHODS: The CDR3 of 24 TCR Vbeta subfamily genes were amplified from peripheral mononuclear cells of a patient with cGVHD after unrelated BMT. RT-PCR was used for detection of the distribution of TCR Vbeta repertoire, and the PCR products were further analyzed by genescan technique for the CDR3 size, to evaluating clonality of the detectable TCR VbetaT cells. RESULTS: Only 8 Vbeta subfamily T cells could be identified in the patient, clonal expansion T cells could be found of Vbeta2, 3, 8 and 13 subfamilies. CONCLUSIONS: The skew distribution and clonal expansion of TCR Vbeta subfamily T cells could be found in the patient. It may relate to the initiation of cGVHD. PMID- 11877012 TI - [Analysis of surface markers of expanded human umbilical cord blood cells in vitro]. AB - OBJECTIVE: To explore the optimal timing for in vitro expansion and transplantation of umbilical cord blood hematopoietic cells. METHODS: Hematopoietic cells from human umbilical cord blood were cultured with G-CSF, GM CSF, rIL-3, rIL-6, SCF and Epo in a long term culture system, the cell cycle and cellular markers were analysed dynamically. RESULTS: CD(3)(+), CD(4)(+)/CD(45RO)(+) and CD(8)(+)/CD(45RO)(+) cells increased at day 3, and reached the highest level at day 7, decreased at day 14, and remained lower level at day 21. CD(4)(+) and CD(8)(+) cells showed no change at day 7, decreased at day 14, and decreased further at day 21. CD(4)(+)/CD(45RA)(+) and CD(8)(+)/CD(45RA)(+) cells decreased after day 3, and decreased further at day 14. HLA-DR(+) cells increased from day 3 to day 7. CD(34)(+) cells increased at day , reached the highest level at day 7, and decreased at day 14. CD(34)(+), CD(34)(+) CD(38)(-), CD(34)(+) CD(38)(+) cells increased 8.8, 7.4 and 8.7 fold at day 7, respectively, as compared with that precultured. The cells in the S phase increased at day 21. CONCLUSIONS: This culture system is able to expand hematopoietic cells of human umbilical cord blood and keeps them intact. The number of hematopoietic cells reached the highest level in the culture from day 7 to day 14, which is the optimal time for transplantation. PMID- 11877013 TI - [Effects of different cryoprotectants on biological properties of hematopoietic cells derived from cord blood]. AB - OBJECTIVE: To search for effective cryoprotectants that can be used in cord blood bank to cryopreserve hematopoietic cells. METHODS: Four kinds of combined reagents were used to cryopreserve cord blood cells. The effects of these cryoprotectants on biological properties of hematopoietic cells were monitored and compared after the cells were refrigerated for 1 to 4 months. The detected biological properties included the number of total nucleated cells (TNC), CD(34)(+) cells and CFCs. RESULTS: The cryoprotectants of Dextran-40 + 10%DMSO resulted in the highest recovery rates, TNC, CD(34)(+) cells and CFCs were 77.8%, 78.5% and 87.2%, respectively. The combination of 10%DMSO + 6% AB serum + 5%HES led to the lowest recovery rates. CONCLUSION: Dextran-40 + 10%DMSO is an ideal cryoprotectant combination for cryopreserving hematopoietic cells in the cord blood bank. PMID- 11877014 TI - [The impact of hepatitis B virus infected patients or donors on the outcome of hematopoietic stem cell transplantation]. AB - OBJECTIVE: To investigate the impact of recipients and/or donors infected with hepatitis B on the outcome of hematopoietic stem cells transplantation (HSCT). METHODS: We analyzed retrospectively the transplantation outcome in 26 of 164 hematological diseases patients who and whose donors were infected with hepatitis prior to transplantation. RESULTS: (1) Three of the 26 patients developed VOD after HSCT, the incidence (11.5%) is significant higher than that in patients and donors who did not infected with HBV (P < 0.05). (2) Two of 5 patients transfused hematopoietic stem cells from HBsAg + donors developed hepatitis B. (3) Four patients with allogeneic stem cell transplantation developed hepatic failure (HF) which occurred during cyclosporin (CsA) being tapered off or withdrawal. The incidence of HF in patients and/or donors infected with HBV (15.4%) is obviously higher than that in patients and donors who did not (0.08%, P < 0.01). (4) In 4 patients with HbeAg +, 2 patients died of HF after allogeneic HSCT, the other 2 patients survived after autologous HSCT. CONCLUSION: Donors and recipients infected with hepatitis B are not contraindication of HSCT, but patients with allogeneic HSCT are in danger to develop LF during CsA being tapered off or withdrawal. PMID- 11877015 TI - [Experimental study on ex vivo expansion of CD(34)(+) umbilical cord blood cells]. AB - OBJECTIVE: To investigate the potential possibility that to expanse ex vivo umbilical cord blood (UCB) cells to an amount sufficient adults transplantation. METHOD: Purified CD(34)(+) UCB cells from 10 fresh UCB samples were cultured for 7 days in IMDM mediums containing 20% FBS and one of the following three combinations: Group A (IL-1beta + IL-3 + IL-6 + G-CSF + Epo + FL), Group B (SCF + IL-1beta + IL-3 + IL-6 + G-CSF + Epo) and Group C (FL + SCF + IL-1beta + IL-3 + IL-6 + G-CSF + Epo). RESULT: (1) There was no significant difference between Groups A and B in expansion of UCB cells. But in group C, the expansion was greater than that in group A (P < 0.01) or group B (P < 0.05). The effects of FL and SCF were synergistic. (2) Over 50 x 10(6) CD(34)(+) cells which are sufficient for adult transplantation were obtained in 30% of UCB samples in group C. (3) The expanded CD(34)(+) UCB cells retained original clonogenic efficiency, the primitive CD(34)(+)CD(38)(-) proportion and the expansion potential were the same as fresh UCB cells. CONCLUSION: The ex vivo expansion of CD(34)(+) UCB cells might provide sufficient hematopoietic stem cells for adult transplantation. PMID- 11877016 TI - [A mutation of platelet glycoprotein I balpha results in defects in its interaction with immobilized vWF]. AB - OBJECTIVE: To study the importance of glycoprotein (GP) I balpha mutation (A156V) in interaction between mutant expressing cell and immobilized vWF under fluid shear stress. METHODS: Mutant GP I balpha cDNA was cloned into the EcoR I site of the mammalian expression vector pDX, mutant cDNA was then transfected into CHO betaIX cells. Human vWF was purified from blood cryoprecipitate by glycine and NaCl precipitation and subsequent separation on sepharose 4B column. The purified vWF was immobilized onto a coverslip. Cell rolling was induced in a parallel plate flow chamber and observed by phase-contrast video microscope. RESULTS: CHO cells expressing GP I b-IX-V complex could adhere to and roll on immobilized vWF. The A156V mutant cells retained the ability to adhere and roll on vWF matrix, but the rolling speed was significantly faster than that of wild type cells, indicating that the off-rate of the ligand-receptor bond between the mutant and vWF was impaired. Binding of monoclonal antibody AN51 to mutant GP I balpha decreased significantly, indicating that the A156V mutation altered the conformation of the N-terminal ligand-binding region of GP I balpha. CONCLUSION: The mutant GP I balpha has a faster off-rate for its interaction with immobilized vWF. The mutant polypeptide adopts an altered conformation in N-terminal ligand binding region of GP I balpha. The parallel-plate flow chamber is an extremely useful system in evaluating interaction between GP I balpha and vWF. PMID- 11877017 TI - [Retroviral-mediated high efficient in vitro expression of human coagulation factor VIII]. AB - OBJECTIVE: To develop a retroviral-mediated high efficient expression system of human coagulation factor VIII. METHODS: The retroviral vector LNC-VIIIBD was generated by cloning a B-domain-deleted FVIII cDNA (760aa - 1639aa) into retroviral vector pLNCX. Several cell lines including NIH3T3, CHO, COS-7 and human hepatic cell line L-02 were infected with viral supernatant from the highest productive PA317 clones. The antigen and procoagulant activity of human FVIII in the cell culture medium were measured by ELISA assay and one-stage method, respectively. RT-PCR was performed for the detection of F VIII BD mRNA. RESULTS: Human FVIII was expressed in all four target cells. The highest expression was observed in NIH3T3, the procoagulant activity of secreted FVIII was up to 1.6 U, and the FVIII antigen was 500 ng by 10(6) cells/ml in 24 hours, respectively. CONCLUSION: The constructed retroviral vector was able to generate high level expression of human FVIII in some cell lines, and it might have potential utility in the gene therapy for Hemophilia A. PMID- 11877019 TI - [The study of beta-fibrinogen gene - 455 G/A, - 148 C/T, 448 G/A polymorphisms and their association with plasma fibrinogen levels]. AB - OBJECTIVE: To analyze the frequency of beta-fibrinogen (beta-Fg) gene - 455 G/A, 148 C/T, 448 G/A polymorphisms and their association with plasma fibrinogen levels in Han nationality in Guangdong Chinese. METHODS: The beta-Fg gene - 455 G/A, - 148 C/T, 448 G/A polymorphisms of 156 individuals were analyzed by restriction fragment length polymorphism (RFLP). Plasma fibrinogen levels were determined by turbidimetry. RESULTS: The frequencies of A(- 455), T(- 148) and A(448) allele were 0.276, 0.285 and 0.272, respectively. There were strong linkages of G(- 455), C(- 148) and G(448), and of A(- 455), T(- 148) and A(448), the correspondence was more than 97%. The plasma fibrinogen levels in the presence and absence of A(- 455) allele were (3.13 +/- 0.74) g/L and (2.89 +/- 0.57) g/L (P < 0.05); of T(- 148) allele were (3.12 +/- 0.73) g/L and (2.89 +/- 0.58) g/L (P < 0.05); and of A(448) were (3.13 +/- 0.74) g/L and (2.89 +/- 0.57) g/L (P < 0.05), respectively. The plasma fibrinogen levels of the three polymorphisms with the mutant gene are significantly higher than that in the wild type. CONCLUSION: The three polymorphisms loci are strong linkage disequilibrium. It suggests that beta-Fg gene - 455 G/A, - 148 C/T, 448 G/A polymorphisms are associated with plasma fibrinogen levels. PMID- 11877018 TI - [Influence of pravastatin on expression of platelet CD(62P) and CD(41) in patients with hypercholesterolemia in vitro and in vivo]. AB - OBJECTIVE: To study in vitro and in vivo effect of hypercholesterolemia (HC) on expression of platelet CD(62P) and CD(41) and their changes after treatment with pravastatin. METHODS: Twenty patients with HC were given 10 - 20 mg/d pravastatin orally for 4 - 8 weeks. Changes of blood cholesterol and expression of platelet CD(62P) and CD(41) were detected by flow cytometry. RESULTS: After 4 and 8 weeks' treatment, blood cholesterol level and platelet aggregation function decreased. The expression of CD(62P) average fluorescence intensity decreased from 31.8 +/- 7.8 to 27.2 +/- 6.9 and 26.8 +/- 4.9; CD(62P) (%) from (31.3 +/- 9.3)% to (26.4 +/- 7.4)% and (25.3 +/- 9.1)% (P < 0.05); CD(41) from 483.2 +/- 263.9 to 348.1 +/ 192.4 and 306.8 +/- 128.0 (P < 0.05), respectively. Similar results were obtained in vitro study. CONCLUSION: Improvement of platelet function and expressions of CD(62P) and CD(41) in HC patients after prevastatin treatment may be resulted from enhancing arteriosclerosis regression. It suggested from the in vitro result that pravastatin might play a direct effect on platelet function. PMID- 11877020 TI - [Preliminary study on the effects of coagulation factor XII on fibrinolysis]. AB - OBJECTIVE: To study the effects of FXII on fibrinolysis in patients with cerebral thrombosis. METHODS: Plasma level of FXII:C, FXII:Ag, FXIIa and beta FXIIa and fibrinolysis activities were examined by ELISA. Screening of FXII gene mutation by MOEA. RESULTS: FXII:C in 22 of 107 patients with cerebral thrombosis decreased, which was similar to the feature of FXII cross-reacting material positive (FXII CRM(+)). There were significant increase in plasma levels of PLG:A and alpha(2)AP:A and decrease in D-dimer, moreover, plasma levels of FXIIa and betaFXIIa were lower in patients than in controls. FXII gene mutation was not found in 22 of 107 patients. CONCLUSION: Decrease of FXII:C may play an important role in cerebral thrombosis by reducing activation of plasminogen. The gene mutation of FXII CRM(+)-like abnormal FXII was different from the known FXII gene mutation. Mutations in the regions of FXII Arg (334) and Arg (353) may be more important for the reducing of FXIIa, beta FXIIa levels and fibrinolysis activities. FXII assay should be included in thrombotic disorder screening. PMID- 11877021 TI - [Hemostatic variations associated with acute leukemia: clinical and prognostic significance]. AB - OBJECTIVE: To study the hemostatic abnormalities and their association with bleeding manifestation and prognosis in acute leukemia (AL) patients. METHODS: A series of hemostatic parameters were determined in 93 cases of AL using ELISA or chromogenic assay. RESULTS: At diagnosis, plasma levels of P-selectin, soluble fibrinomonomer complex (SFMC), thrombomodulin (TM), tissue plasminogen activator, D-dimer (D-D) were significantly elevated, but protein C antigen (PC:Ag), plasminogen (PLG), alpha(2)-antiplasmin (alpha(2)-PI) and plasminogen activator inhibitor (PAI) were decreased, and fibrinogen (Fg), PC activity (PC:A) and protein S (PS) were in normal range. All the above abnormalities returned to normal after complete remission (CR) except for PC:A and PC:Ag. Platelet counts, Fg, SFMC, PLG, alpha(2)-PI and D-D were significantly associated with hemorrhagic manifestations. TM, PS and PAI were related to prognosis. CONCLUSION: There existed the consumption of platelets as well as the activation of coagulant, anticoagulant and fibrinolytic systems in AL, which was greatly improved after CR. PMID- 11877022 TI - [Detection of soluble endothelial protein C receptor (sEPCR) in patients with CHD, DM and SLE]. AB - OBJECTIVE: To evaluate the clinical significance of plasma soluble endothelial protein C receptor (sEPCR) level in patients with coronary heart disease (CHD), diabetes mellitus (DM) and systemic lupus erythematosus (SLE). METHODS: ELISA of antibody-sandwiched principle was used to detect sEPCR in 34 patients with CHD, 42 with DM-II, 63 with SLE and 20 normal controls (NC). RESULTS: The plasma level of sEPCR in patients with DM was higher than that in NC group (P < 0.05). The sEPCR level in SLE patients with thrombosis and non-thrombosis are higher than that in NC group (P < 0.005 and P < 0.001, respectively). There is no significant difference between the two groups of SLE (P > 0.05), and between the CHD and NC groups. CONCLUSION: sEPCR level increased in DM and SLE, it seems related to the process of inflammatory reaction and damage of endothelial cell. PMID- 11877023 TI - [Changes of vWF in vascular endothelial cells with acute hypoxia]. AB - OBJECTIVE: To investigate changes of vWF in vascular endothelial cells with acute hypoxia and assess its significance in the study of vascular injury and hypercoagulability. METHODS: vWF in vascular endothelial cells and in blood were measured by immunohistochemical staining image analysis and ELISA technique. Fibrin was displayed by phosphotungstic hematoxylin staining. RESULTS: The amount of vWF in vascular endothelial cells decreased progressively, while in blood increased progressively with hypoxia. Fibrin prominently increased with hypoxia. CONCLUSION: Acute hypoxia caused severe injury of vascular endothelial cells which resulted in release of vWF into blood from endothelial cells. The vWF amounts in blood may reflect the hypercoagulability, and may serve as a useful indicator of vascular injury and hypercoagulability. PMID- 11877024 TI - [A novel mutation of the ALAS2 gene in a family with X-linked sideroblastic anemia]. AB - OBJECTIVE: To confirm the mutation of ALAS2 gene is the cause of sideroblastic anemia in a family. METHODS: Polymerase chain reaction (PCR) was used to amplify the microsatellite DXS 991, DXS 1199 in the chromosome Xp11.22 linked gene ALAS2 and haplotype analysis was performed in a kindred with 2 patients and 7 normal members. All cDNA encoded regions in the ALAS2 gene of the patients and their normal siblings were cloned, sequenced and compared. RESULT: Both brother patients had the same allele of ALAS2 and their normal siblings did not. The mutation in the patients' ALAS2 gene was exon 5 A523G, causing threonine to alanine; and exon 3 T372C, leucine to proline. The latter located in the splicing region, its significance is not clear. CONCLUSION: The pathogenesis of this kindred of X-linked sideroblastic anemia (XLSA) involved a novel mutation in ALAS2 exon 5. PMID- 11877025 TI - [A report of two Chinese familial Budd-Chiari syndrome]. AB - OBJECTIVE: To investigate the etiology of two Chinese familial Budd-Chiari syndrome (BCS). METHOD: Four patients with familial BCS (from A and B families), and the other 41 family members were detected by angiography, ultrasound Dopler, etiology analysis and Factor V Leiden (FvL) mutation analysis. RESULT: Four BCS patients were proved by angiography, 2 by ultra sound Dopler in family A. Ten members in family A were varicosis in low extremeties. FvL mutation was detected in 4 of 6 patients and 2 normal family members. A II(2), A III(7, 11, 15,) B II(10) and B III(5) had FvL mutation. The FvL mutations were compatible with Mendel hereditary law. CONCLUSION: FvL mutation may be one of main risk factors and varicosis in low extremeties may be another risk factors for familial BCS. PMID- 11877026 TI - [The preliminary study on the gene mutations of sixty patients with G6PD deficiency in Yunnan province]. AB - OBJECTIVE: To analyze the types and distribution of glucose-6-phosphate dehydrogenase (G6PD) gene mutations in Yunnan province. METHODS: By using the natural primers or mis-matched primers mediated polymerase chain reaction (PCR) followed by endonuclease digestion methods, six types of G6PD gene mutations, G1388A, G1376T, C1024T, G392T, C592T and A95G were in sixty patients with G6PD deficiency. RESULT: Six types of mutation were revealed: 28.3% (17 of 60) were G1388A, 11.7% (7 of 60) G1376T, 6.7% (4 of 60) C1024T, 1.7% (1 of 60) G392T, 1.7% (1 of 60) C592T, 1.7% (1 of 60) A95G and 48.3% (29 of 60) unidentified. C592T mutation is the first report in Yunnan people. CONCLUSIONS: The G6PD gene in Yunnan people is heterogeneous. Research of G6PD gene mutation types may provide some useful data for clinical diagnosis and prevention of G6PD deficiency, and for human genetic study. PMID- 11877027 TI - [Separation and enrichment of fetal nucleated red blood cells from maternal blood for non-invasive prenatal gene diagnosis]. AB - OBJECTIVE: In order to develop a non-invasive technique for prenatal gene diagnosis. METHODS: Peripheral blood mononuclear cells (MNCs) were separated by single density gradient Histopaque 1.077 from 25 pregnant women with gestations between 8 - 36 weeks. The fetal nucleated red blood cells (NRBCs) were enriched from the MNCs by positive selection using Dynabeads M-450 CD(71) or negative selection using Dynabeads M-450 CD(45). The enriched NRBCs were identified by anti-gamma-biotin or anti-zeta-biotin antibodies. Globin gene of NRBCs from fetuses with risk of beta-thalassemia major were amplified with nested PCR followed by reverse dot blot hybridization for gene diagnosis. RESULTS: NRBCs stained by anti-gamma-biotin or anti-zeta-biotin antibodies could be found in the peripheral blood samples of the 25 pregnant women. Three out of 5 fetuses with risk of beta-thalassemia major were successfully diagnosed using these NRBCs. CONCLUSION: Fetal NRBCs in maternal circulation can be isolated and enriched by single gradient density Histopaque 1.077 followed by magnetic activated cell sorting. Nested PCR can amplify DNA for gene diagnosis from no less than 20 NRBCs. PMID- 11877028 TI - [Diagnosis of PNH by detecting the resistance of RBCs to the hemolytic effect of hec toxin secreted by Aeromonus hydrophila]. AB - OBJECTIVE: To study the feasibility of diagnosing paroxysmal nocturnal hemoglobinuria (PNH) with hec toxin (secreted by Aeromonus hydrophila) test. METHODS: The crude hec toxin was extracted from the culture medium of Aeromonus hydrophila by precipitating with saturated (NH(4))(2)SO(4) and then purified through DEAE52. This crude hec toxin was used to act on red blood cells (RBCs) from patients with PNH, non-PNH anemias, and normal persons. Absorbance at 630nm was measured to quantitate the extent of hemolysis. Hec treated and untreated RBCs were both stained with anti CD(59) monoclonal antibody and FITC labelled goat-anti-mouse IgG. The percentage of CD(59)(+) cells was detected by flow cytometry. RESULTS: After hec toxin treatment, RBCs from PNH patients showed resistance to the toxin hemolysis, which was negatively related to the CD(59)(+) cells percentage, while RBCs from non-PNH were lysed totally. CONCLUSION: RBCs from PNH have obvious resistance to the hemolytic effect of hec toxin, and the percentage of remained unhemolytic cells reflect the severity of PNH. Resistance detection of RBCs to hec toxin can be used for the diagnosis of PNH. PMID- 11877029 TI - [Lymphocyte subsets analysis and glycosylphosphatidylinositol determination in patients with PNH]. AB - OBJECTIVE: Absolute number of lymphocyte subsets and the proportion of glycosylphosphatidylinositol (GPI)-deficient clones among T, B and NK cells were determined. METHODS: By using multicolor flow cytometry with two and three color labelled monoclonal antibodies, absolute number of lymphocyte subsets and the proportion of CD(48), CD(55), CD(59)-deficient T, B and NK cells were analyzed. RESULTS: Lymphocyte subsets were found abnormal in all patients, low absolute number of natural killer cells and B cells were obvious. In most patients, GPI deficient T, B and NK cells were detectable. The proportion of GPI-deficient clones in T, B and NK cells were 78.7%, 53.4% and 30.5%, respectively. There was significant difference among T, B and NK cells (P < 0.01). In two continuous remission cases, no GPI-deficient clones were found in erythrocytes, but there were GPI-deficient T and B cells, and in a 3 year remission there were still GPI deficient NK cells. CONCLUSIONS: In PNH, similar to other blood cells, lymphocytes also had GPI deficiency, and GPI-deficient T and B cells could exist for a long time. NK cells of PNH patients are the predominant, GPI-deficiency cells with a high frequency (93.3%) and high proportion (78.7%). PMID- 11877030 TI - [Quantitative determination of EAIgG by ABC-ELISA and its clinical significance]. AB - OBJECTIVE: To explore a method for diagnosis of autoimmune hemolytic anemia (AIHA) and evaluate its clinical significance. METHODS: Erythrocyte associated IgG (EAIgG) was quantified in 60 healthy people, 44 patients with warm antibody AIHA (WAIHA), 108 non-immune anemia and 44 other immune diseases (total 256 cases) by using ABC-ELISA were estimated. RESULTS: EAIgG in WAIHA patients were significantly higher than those in other groups (P < 0.01). In the 44 WAIHA patients, 38 were Coombs positive and 6 Coombs negative. EAIgG in Coombs negative was significantly high. The dynamic changes of EAIgG in 12 WAIHA patients were observed during treatment. When patient's condition got better, Hb and RBC increased and EAIgG decreased. CONCLUSION: Quantitative detection of EAIgG is an index in the diagnosis of WAIHA, especially in AIHA with negative Coombs' test. It is also a useful factor for evaluating therapeutic effect and prognosis in WAIHA. PMID- 11877031 TI - [The development of immune-mediated aplastic anemia in mice can be blocked by cimetidine]. AB - OBJECTIVE: To investigate the effect of H(2) receptor antagonist cimetidine on the development of immune-mediated aplastic anemia (AA) in mice. METHODS: The immune-mediated aplastic anemia mice model was induced by irradiation and half matched allogeneic lymphocytes transfusion (ALT). The mice were divided into 3 groups: the irradiation alone control group, the irradiation + ALT group (AA group) and the irradiation + ALT + cimetidine group. In cimetidine group, each mouse was intraperitoneally injected cimetidine at different doses once a day for 8 days. On day 14, the bone marrow histology was examined. RESULTS: In 0.5 mg, 1.0 mg and 2.0 mg x (10g)(-1) x d(-1) cimetidine groups, the percentages of hematopoietic tissue volume in bone marrow were (46.58 +/- 20.41)%, (51.88 +/- 24.94)% and (59.12 +/- 32.48)%, respectively, and all much higher than that in AA group [(19.99 +/- 10.98)%, P < 0.01], but were not different from that in irradiated control group [(53.81 +/- 31.72)%, P > 0.05]. CONCLUSION: The development of immune-mediated aplastic anemia in irradiated mice by half-matched allogeneic lymphocytes transfusion was blocked by H(2) receptor antagonist cimetidine. PMID- 11877032 TI - [A comparative study on the expressions of IL-4, IFN-gamma and TNF-alpha in BMMNC of acute and chronic aplastic anemia patients]. AB - OBJECTIVE: To detect the induced levels of IL-4, IFN-gamma and TNF-alpha in the supernatant of bone marrow mononuclear cells (BMMNC) and compare the difference of immune status between acute (SAA) and chronic (CAA) aplastic anemia patients. METHODS: Concentrations of IL-4, IFN-gamma and TNF-alpha in PHA-P-induced BMMNC supernatants were determined by ELISA assay in 11 SAA and 13 CAA patients as well as 16 controls. Concentration differences between the two groups were compared. RESULTS: (1) The IFN-gamma and TNF-alpha levels in AA patients studied were much higher than that in controls, and IL-4 levels were normal in SAA group but elevated in CAA group. (2) TNF-alpha levels were comparable between the two AA groups, but both IL-4 and IFN-gamma levels were significantly different between them. CONCLUSION: Enhanced cellular immunity seems to play an important role in the pathogenesis of SAA, and enhancement of both cellular and humoral immunity might contribute to the pathogenesis of CAA. PMID- 11877034 TI - [Long-term expression of MHC-I gene in recipient mice]. AB - OBJECTIVE: To explore the possibility of long-term expression of transferred MHC I gene in recipient mice. METHODS: MHC-I gene of the donor mouse (C(57)BL/6) was transferred into the recipient (BALB/c) hematopoietic cells by retrovirus mediated gene transfer technique, and the gene transferred cells were reinfused into the recipient mice. PCR, RT-PCR and FACS were used to assay the long-term expression of MHC-I gene in the recipient mice. RESULTS: The gene transferred mice hematopoietic cells and their G418-resistant CFU-GM were demonstrated to be able to express the MHC-I gene. The cells proliferation and differentiation had no change. The MHC-I gene could be expressed for a longer period in the recipient mice. CONCLUSION: This experiment makes ready for the further research on inducing transplantation tolerance through MHC-I gene transfer. PMID- 11877033 TI - [Umbilical cord blood stem cells transplantation from an unrelated donor into an adult with severe aplastic anemia]. AB - OBJECTIVE: To evaluate the availability of the treatment of adult severe aplastic anemia with unrelated allo-cord blood stem cell transplantation (CBSCT). METHODS: HLA-matched unrelated cord blood transplantation has been successfully performed for an adult severe aplastic anemia patient. A unit of cord blood provided by Guangzhou Cord Blood Bank containing 1.89 x 10(7)/kg mononucleated cells, 1.8 x 10(4)/kg CFU-GM and of CD(34) positive cells was 0.009. The patient was conditioned with CTX (60 mg/kg) and anti-lymphocyte globulin (ALG, 120 mg/kg). GVHD prophylaxis consisted of both MTX and CsA. The CsA had been given for 8 months. RESULTS: The lowest ANC was 0.6 x 10(9)/L post-transplantation. The patient achieved an ANC of greater than 0.5 x 10(9)/L at 10 days, and the platelet of greater than 50.0 x 10(9)/L at day 20 after transplantation. He developed Grade 1 GVHD in the tenth month after grafting. Microsatellite DNA fingerprinting indicated a stable donor-recipient mixed chimerism, whilst the circulating red cells remained host origin. CONCLUSION: It is the first report in China on successful treatment of adult severe aplastic anemia by unrelated allo CBSCT. PMID- 11877035 TI - [Expression of lipofect AMINE mediated human GM-CSF eukaryotic expressing vector in HFCL cells]. AB - OBJECTIVE: To construct an eukaryotic expressing vector-pIRES1neo/hGM-CSF and express it in human bone marrow stromal cell line HFCL. METHODS: Human granulocyte/macrophage colony-stimulating factor cDNA (hGM-CSF cDNA, 751 bp) was inserted into an effective eukaryotic expressing vector-pIRES1neo which contains the human cytomegalovirus (CMV) major immediate early promoter/enhancer and the internal ribosome entry site (IRES) of the encephalomyocarditis virus (ECMV). HFCL cells were transfected with the recombinant vector pIRES1neo/hGM-CSF by liposome-mediated gene transfer method. Integration of hGM-CSF in the genome, transcription of its mRNA and expression of its protein in the transfected HFCL cells were assayed by Southern blot, Northern blot, ELISA and hGM-CSF dependent cell line TF-1. RESULTS: hGM-CSF cDNA was integrated into HFCL genome successfully, hGM-CSF mRNA was transcripted and hGM-CSF protein was expressed of (56.9 +/- 0.7) ng/10(6) cells by ELISA and (6.56 +/- 0.16) x 10(3) U/10(6) cells per day by TF-1 cell assay in the supernatant. CONCLUSION: The recombinant vector is proved to be stably expressed in HFCL cells and the biological activity of hGM CSF was detectable in the supernatant of the transfected cells. PMID- 11877037 TI - [Construction of membrane-anchored interleukin-6 receptor beta subunit mutant and its effect on IL-6 signal transduction]. AB - OBJECTIVE: To investigate whether the membrane-anchored gp130 mutant containing the second and the third fibronectin type III modules of the extracellular region could still transduce IL-6 signal. METHOD: The membrane-anchored gp130 mutant cDNA was amplified by the overlap extension PCR and inserted into the mammalian expression vector pRc/RSV. The mutant was proved to be effectively expressed in SKO-007 cells and Jurkat cells by dot blot hybridization. The mutant's effects on IL-6 signaling in these two kinds of cells were detected by electrophoresis mobility shift assay (EMSA). RESULTS: In SKO-007 cells, the mutant could enhance the activated APRF, but the effect was not as strong as that of wide-type gp130. IL-6 could not activate nuclear factors in Jurkat cells. After the expression vectors pRc/RSVgp130 and pRc/RSV were transfected into Jurkat cells, IL-6 could activate APRF, but could not do NF-IL6. Similar to the results in SKO-007 cells, the effects of the mutant was also weaker than that of wild-type gp130. CONCLUSION: Although the mutant could still transduce IL-6 signal, it is not as efficiently as the wild-type gp130. It suggested that the Ig-like domain of gp130 also played an important role in IL-6 signal transduction. PMID- 11877036 TI - [Experimental studies on the characteristics of suicide gene system HSV-tk/GCV and its effect on dendritic cells]. AB - OBJECTIVE: To study the characteristics of suicide gene system of the herpes simplex virus thymidine kinase (HSV-tk) gene in tumor cells and explore the apoptosis phenomena in this system and its effect on dendritic cells (DC). METHOD: HSV-tk was transduced into the breast cancer cell line MCF-7 cells with a retrovirus vector. Apoptosis was examined by electro-microscopy and FACS. DCs were induced from cord blood CD(34)(+) cells with a combination of cytokines (GM CSF, TNF-alpha, SCF and FL) and (3)H-TdR was adopted to determine the proliferation of T cells. RESULTS: In vitro experiment showed a dose-dependent cell killing of the HSV-tk gene transducted cells on ganciclovir (GCV) treatment. The apoptosis rate increased to 31.3% from the control group of 19.3%. Apoptosis is the important mechanism in bystander effect of HSV-tk/GCV system. The CD(1a)(+) and HLA-DR(+) cells in the induced DC were of (27.18 +/-1.56)% and (93.7 +/- 1.0)%, respectively. Co-incubation of DC with apoptotic cells further stimulated the proliferation of T cells. CONCLUSION: It suggested that HSV-tk/GCV system, alone or combined with radiotherapy, be a promising suicide gene therapy for cancer patient and an approach to enhancing DC function. PMID- 11877038 TI - [Construction, eukaryotic expression and biological activities of a recombinant human single chain interleukin-12 fusion gene]. AB - OBJECTIVE: To explore the feasibility of recombinant human interleukin 12 (IL-12) with biological activities by molecular biological techniques. METHOD: Both p40 and p35 subunits cDNA of human IL-12 were cloned from mRNA extracted from NC-37 cell line by using RT-PCR, and the fusion gene (p40-linker-p35) of recombinant human single chain IL-12 (rhscIL-12) was constructed by using a polypeptide linker (Gly(4)Ser)(3). rhscIL-12 eukaryotic expressing vector pcDNA3.1 (+)-hscIL 12 was constructed by inserting the rhscIL-12 fusion gene into pcDNA3.1 (+) eukaryotic expressing plasmid. COS-7 cells were transfected with pcDNA3.1 (+) hscIL-12 plasmid. RESULT: A stable rhscIL-12 expressing cell line COS-rhscIL-12 was obtained by G418 selection. Western blot showed the presence of a 70 x 10(3) band of the fusion protein, which specifically bond to mouse-anti-human IL-12 monoclonal antibody. The assays of biological functions showed that the fusion protein had strong bioactivities in stimulating the lymphocyte proliferation, enhancing the NK cell cytotoxicity and increasing the IFN-gamma production. CONCLUSION: The constructed rhscIL-12 fusion gene could express biologically functional rhscIL-2 fusion protein. PMID- 11877039 TI - [The sensitivity to apoptosis is enhanced in U937-ASPI-3K cells which stably express antisense ATM/PI-3K]. AB - OBJECTIVE: To analyse the characteristics of apoptosis sensitivity in U937-ASPI 3K cells which is stably expressing anti-sense ATM/PI-3K. METHOD: Annexin V-PI apoptosis detection kid and flow cytometry were used to determine apoptosis rate, Western blot to the expression level of cytochrome C protein and Bcl-2 protein. RESULTS: U937-ASPI-3K cells had a lowered threshold for triggering apoptosis in response to low dose irradiation. Cell apoptosis rate was 28% and (53.0 +/- 5.4)%, respectively, at 8 and 48 hours after 1.5 Gy (137)Cs irradiation, while in U937-pZEOSV2 (+) control group it was 4.2% and (11.0 +/- 3.3)%, respectively. Cytosolic cytochrome C protein band was gradually intensified in U937-ASPI-3K in 4 - 8 h after irradiation, but was not detected in control group. Bcl-2 protein gradually decreased in U937-ASPI-3K in 4 - 8 h after irradiation, but didn't change obviously in control group. CONCLUSION: Cell line U937-ASPI-3K acquired a phenotype with a lowered threshold for triggering apoptosis. It demonstrated that selective inactivation of ATM gene might be of considerable value in tumor treatment. PMID- 11877040 TI - [P15(INK4B) gene methylation in malignant hematopoietic diseases]. AB - OBJECTIVE: To study the effect of operative region hypermethylation gene in human malignant hematopoietic tumors. METHODS: The abnormal methylation rate of P(15)(INK4B) gene 5'CpG island in 68 cases of malignant hematopoietic tumor samples were determined by methylation specific PCR using bisulfite modified DNA. RESULTS: The methylation rates of P(15)(INK4B) were 84%, 0, 50% and 75%, respectively, for 25 cases of acute myeloid leukemia (AML), 15 chronic myeloid leukemia (CML), 16 myelodysplastic syndrome (MDS) and 12 multiple myeloma (MM). P(15)(INK4B) gene was frequently methylated in patients with high risk MDS and early stage of MM. CONCLUSION: Hypermethylation of P(15)(INK4B) gene is one of the main causes of its inactivation. Hypermethylation of CpG island was closely related to the development of malignant hematopoietic diseases. PMID- 11877041 TI - [Multivariate analysis of prognostic factors in Philadelphia chromosome-positive chronic granulocytic leukemia]. AB - OBJECTIVE: To investigate the prognostic factors of the patients with Philadelphia chromosome-positive chronic granulocytic leukemia (CGL). METHODS: Clinical and laboratory parameters of 209 CGL patients at diagnosis were analyzed in relation to survival. COX regression analysis were used to identify the statistically significant prognostic factors. RESULTS: The median survival time was 57 (3 - 179) months, and 5 year survival rate was 48%. From the COX regression model, the four main poor prognostic factors were as follows: percentage of bone marrow blasts (>or= 0.04), percentage of peripheral blood blasts and promyelocytes and presence of erythroid precursors in peripheral blood. According to the prognostic scores, three groups of CGL were identified. The low-risk group was with a median survival time of 76 months, the intermediate risk group with 56 months, and the high-risk group with 31.5 months (P < 0.01). The 5 year survival rate was 70.42%, 46.30% and 18.18% (P < 0.01), respectively. CONCLUSIONS: A prognostic scoring system suitable for Chinese CGL patients was proposed. PMID- 11877043 TI - [Correlation of cell apoptosis induction with expression of human beta5 integrin on hematopoietic cells]. AB - OBJECTIVE: To investigate the function of alpha(v)beta5 integrin in hematopoietic cells. METHODS: Beta5 integrin cDNA was expressed in K562 cells through a retroviral vector system. The changes of the alpha(v)beta5 and alpha(v)beta3 integrins expression in apoptosis and differentiation induced by serum depletion were observed. RESULT: The beta5 integrin cDNA failed to express in K562 cells after the introduction of the beta5 integrin retrovirus vector pG beta5CHT. Imbalanced expression of alpha(v)beta3 and alpha(v)beta5 integrins occurred during apoptosis and differentiation induced by serum depletion. Treatment of hematopoietic cells with anti-alpha(v)beta5 monoclonal antibody inhibited the cells apoptosis. CONCLUSION: Overexpression of the alpha(v)beta5 integrin cDNA in hematopoietic cells was associated with the inhibition of cell proliferation and apoptosis. PMID- 11877042 TI - [Regulation of c-myc expression in leukemic cell line Meg-01 by the death domain of Fas gene]. AB - OBJECTIVE: To investigate the mechanism of disordered proliferation of leukemic cells and the proliferation inhibition by the Fas death domain (FASDD) in human leukemia cell line Meg-01. METHODS: The chimerical receptors (Fas/190, Fas/130) were constructed by Fas cytoplasmic domain with the leukemic inhibitory factor (LIF) receptor subunits gp190 and gp130, another chimerical receptor was constructed by replacing the gp130 in Fas/130 with Fas death domain (FAS/130f). The chimerical receptors were separately expressed on the membrane of Meg-01, and activated with anti-Fas antibodies to induce the oligomerics of the cytoplasmic regions (190cyt-190cyt-190cyt, 130cyt-130cyt-130cyt or FASDD-FASDD-FASDD) for initiating the intracellular signal transduction. c-myc expression level was assayed by means of immunoblotting and immunobiochemistry. RESULTS: Increased c myc expression and cell proliferation were observed in the group of Fas/130 as compared with the other two groups. CONCLUSION: The cytoplasmic domain of gp130 might involve the induction of c-myc expression and the cell proliferation of Meg 01 cell. The Fas death domain may be a medium in the apoptosis induction against the effect of gp130. PMID- 11877044 TI - [Preliminary study on CTL enhancement induced by IL-4 gene modified HL-60 cells]. AB - OBJECTIVE: To explore the mechanism of enhancing killing activity of tumor cell specific cytotoxic T lymphocyte (CTL) by IL-4 gene modified tumor cells (IL 4mTC). METHODS: IL-4 gene was introduced into HL-60 cells through retroviral vector PL-IL-4-SN. Wild and IL-4 mTC were used to induce CTL responses, and cell surface molecules were assayed by flow cytometry. RESULTS: The killing activity of tumor cell-specific CTL in IL-4 mTC group was increased from 11.8% to 77.2%, about sevenfold higher than that induced by wild HL-60 cells (P < 0.01). High level expression of MHC class II antigens as well as B7-1 and ICAM-1 molecules was observed in IL-4 mTC. The expression of MHC class I antigen was not affected by IL-4 gene modification. The expression of cell surface molecules induced by IL 4 mTC was completely abrogated by anti-IL-4 McAb. A significant increase of IL-2 secretions was detected during IL-4 mTC inducing CTL responses. IL-2 secretion and CTL response were inhibited by anti-IL-4 or anti-surface molecule McAbs. CONCLUSION: IL-4 gene modification might enhance the tumor cell-specific CTL killing activities by inducing cell surface molecules expression and IL-2 secretion. PMID- 11877045 TI - [Application of triple-color flow cytometry for minimal residual disease detection in acute B-lymphoblastic leukemia]. AB - OBJECTIVE: To set up a sensitive triple-color flow cytometric assay (FCM) for detection of residual leukemia cells. METHODS: Application of triple-color flow cytometry to the patients for detecting CD(10)(+)CD(22)(+)CD(45)(- or dim) leukemic cells in acute B lymphoblastic leukemia (B-ALL). RESULTS: Leukemic cells in B-ALL patients' bone marrow (BM) consistently showed the CD(10)(+)CD(22)(+)CD(45)(- or dim) phenotype. FCM analysis could detect leukemic cells at the level of 1 per 10(4) cells. The CD(10)(+)CD(22)(+)CD(45)(- or dim) cell population was absent in normal BM, regenerating BM, and BM from leukemia patients in continuous complete remission. This assay could identify residual leukemic cells in 50 days after starting induction chemotherapy. Furthermore, a gradual increase of leukemic cell population could be monitored by this assay before morphological confirmation of BM relapse. CONCLUSION: Triple-color FCM was a sensitive method for detecting BM CD(10)(+)CD(22)(+)CD(45)(minus sign or dim) leukemic cells and could be an additional useful tool to monitor MRD in B-ALL patients. PMID- 11877046 TI - [p15 gene expression in acute lymphoblastic leukemia cell line Molt4 induced by arsenic trioxide]. AB - OBJECTIVE: To explore the correlation between arsenic trioxide and gene methylation in an acute lymphoblastic cell line. METHODS: Methylation of p15 gene in Molt4 cell line was detected by polymerase chain reaction (PCR) using methylation specific primer (MSP) and the expression of this gene after arsenic trioxide treatment was detected by reverse transcriptase PCR (RT-PCR). The cell cycle and cell growth curve were also observed by flow cytometry. RESULTS: p15 gene failed to express in molt4 after methylation. The expression was recovered, cell growth was inhibited, and G(1) cell cycle arrest was observed, when the cells exposed to arsenic. CONCLUSION: Arsenic trioxide could activate the expression of p15 gene and reverse cell cycle negative regulation. PMID- 11877047 TI - [GATA-2 gene expression in leukemia patients and its significance]. AB - OBJECTIVE: To investigate GATA-2 gene expression in leukemia patients and its clinical significance. METHODS: GATA-2 gene transcripts were detected by reverse transcriptase polymerase chain reaction (RT-PCR), and bcr/abl or PML/RAR alpha transcripts were detected in patients with chronic myeloid leukemia (CML) and acute promyelocytic leukemia (APL), respectively. RESULTS: The GATA-2 transcripts were revealed in 93% of acute myeloid leukemia (AML), in 70% of acute lymphoblastic leukemia (ALL), and 83% of CML patients, but not in normal bone marrow and peripheral blood cells. GATA-2 expression in remission patients was similar to newly diagnosed or relapsed patients. In post-transplantation patients, GATA-2 expression was significantly decreased. In these cases GATA-2 transcripts could be detected with bcr/abl fusion gene disappeared in 2 of 12 CML, whereas neither GATA-2 nor PML/RARalpha transcripts could be detected in APL. CONCLUSION: GATA-2 transcripts was highly expressed in leukemia patients and had no change in remission but significantly decreased in post-transplantation patients. Detection of GATA-2 expression would give information about the residual leukemia stem cells. PMID- 11877048 TI - [The induction of high-level expression of a novel VL-30 like gene during the chemical-inducing erythroid differentiation of murine erythroleukemia cells (MEL)]. AB - OBJECTIVE: To study the gene expression during the chemical inducing erythroid differentiation of murine erythroleukemia cells (MEL). METHODS: Differential display assay was used to analyze the gene expression before and after the induction differentiation of MEL cells by DMSO and hemin; and then, the flanking sequences of one of the cDNA fragment was amplified by modified rapid amplification of cDNA end (RACE) method, Northern blot analysis was adopted to characterize the expression of this gene. RESULT: The expression of a new transcript similar to VL-30 retrotransposon family increased significantly during the chemical inducing erythroid differentiation in MEL cells. The cloned cDNA is 1883bp, and it is highly homologous to the internal region of murine BVL-1 (1,955 4,000 nt). At least three transcripts in MEL cells were detected by Northern hybridization and all of them increased after the induction. CONCLUSION: A new VL 30-like gene is identified for the first time in MEL cells during chemical inducing erythroid differentiation. PMID- 11877049 TI - [Observation of long-term therapeutic outcome in chronic granulocytic leukemia]. AB - OBJECTIVE: To analyse the long-term outcome of various therapeutic protocol for the treatment of chronic granulocytic leukemia (CGL). METHODS: Median duration of chronic phase, survival and transformation of 331 CGL patients treated with different project were retrospectively analyzed. RESULT: Longer chronic phase and survival and lower percentage of transformation were found in patients using mesoindigo as maintaining treatment as compared with those using busulfan as maintaining treatment. The effect of meisoindigo and hydroxyurea showed no significant difference. These two drugs were both superior to bulsufan. Combination chemotherapy was not superior to meisoindigo, hydroxyurea or busulfan alone. Interferon had the best therapeutic effectiveness for the treatment of CGL. CONCLUSION: Busulfan was not suitable for maintaining treatment. Meisoindigo or meisoindigo combined with hydroxyurea was good choices for maintaining treatment. Meisoindigo and hydroxyurea had better effect in combination than alone. Interferon could significantly prolong the chronic period and survival period of CGL patients. PMID- 11877050 TI - [Autologous dendritic cells eliciting cytotoxicity of bone marrow cells against chronic granulocytic leukemia]. AB - OBJECTIVE: To investigate the activity of bone marrow cells activated by autologous dendritic cells (DC) to mediate cytotoxicity against chronic granulocytic leukemia (CGL) cells. METHODS: DC were separated from bone marrow mononuclear cells (BMMNC) of two CGL patients in hematological remission and harvested after 3 days of culture in IMDM containing autologous plasma, rhGM-CSF and rhTNFalpha at 37 degrees C, 5% CO(2) humidified atmosphere. BMMNC obtained from the patients were divided into 3 groups to set up Dexter systems: the control group, rhIL-2 containing, and the third group having DC added at day 4. After 10 days of culture, non-adherent cells were harvested and the changes of immunological phenotype and the percentage of P210 positive cells were analyzed. The cytotoxicity were assayed with two-colour flow cytometry. The non-adherent cells from all the 3 systems served as effector cells, those from control system as target cells. RESULTS: The cytotoxic activity against target cells was greater in the DC-activated effector cells than that in rhIL-2-activated ones. The percentages of death cells in target cells were 63.12% versus 42.59% (case 1) and 61.60% versus 21.46% (case 2), respectively. In addition, there was a marked increase in the death cell percentage in the DC-activated effector cells themselves after incubation with target cells. This phenomenon was not found in the rhIL-2-activated effector cells. The percentage of P210 positive cells was significantly lower in non-adherent cells after 10 days of culture in Dexter system, comparing with that in non-cultured BMMNC. The least P210 positive cells were found in those cultured with DC and the less in those with rhIL-2. CONCLUSION: Autologous DC were able to activate bone marrow cells to generate cytotoxicity against CGL cells. Their effect was greater than that of rhIL-2. These activated bone marrow cells might mediate graft versus leukemia effect in vivo. PMID- 11877051 TI - [Dynamic monitoring of Ph positive cells during early post-transplant period in chronic granulocytic leukemia by interphase FISH]. AB - OBJECTIVE: To assess the effectiveness of transplantation and observe the kinetics of residual leukemic cells during early post-transplantation period in chronic granulocytic leukemia (CGL). METHODS: Ten normal controls, seven untreated CGL patients and eleven CGL patients after allogeneic bone marrow transplantation (allo-BMT) or peripheral blood stem cell transplantation (PBSCT) were studied by interphase fluorescence in situ hybridization (I-FISH) with the FITC fluorescein labeled BCR probe. Conventional cytogenetic analysis (CCA) and reverse transcriptase-polymerase chain reaction (RT-PCR) were also employed. RESULTS: The normal cutoff was defined as less than 6.68% (x +/- 3s) from the analysis of normal controls. The reproduciblity of FISH method was confirmed by serial dilutions by mixing Ph positive cells and normal cells. Forty specimens from 11 CGL patients at 1 week to 6 years post-BMT were investigated after PBSCT and the results showed that (1) FISH and RT-PCR could accurately evaluate Ph (+) cells during early transplantation period (within one month), but RT-PCR was more sensitive in six months after BMT. (2) Residual Ph (+) cells tended to be linear decreasing with the time after transplantation and fell in normal range in most samples three months later. The mean time of normaligation was 57 days (20 to 170 days). (3) One case relapsed and received a second transplantation. FISH presented more precise data for predicting relapse as compared to CCA and RT-PCR. CONCLUSIONS: I-FISH as a useful tool can not only offer more precise assessment of residual leukemic cells compared with CCA and RT-PCR, but also provide dynamic monitoring during early post-BMT period to evaluate the transplantation outcome and predict relapse. PMID- 11877052 TI - [Inhibition of proliferation and induction of apoptosis by simvastatin in K562 leukemic cell line]. AB - OBJECTIVE: To investigate the anti-apoptotic mechanism and explore approach to inhibiting proliferation and inducing apoptosis of chronic myclogenous leukemia (CML) cells. METHODS: K562 cell line was used to evaluate the effects of simvastatin, an inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase, and the combination of simvastatin with chemotherapeutic agents on the proliferation and apoptosis of CML cells. RESULTS: Simvastatin could significantly inhibit proliferation and induce apoptosis of K562 cells, and could increase the sensitivity of K562 cells to chemotherapeutic agents. Addition of mevalonate, the immediate product of HMG-CoA, could completely reverse this effect. CONCLUSION: Simvastatin inhibited proliferation and induced apoptosis of K562 cells through inhibiting the metabolic pathway of mevalonate. It is promising that HMG-CoA reductase inhibitors may be an effective chemotherapeutic approach to the treatment of CML. PMID- 11877053 TI - [Combination of mycophenolate mofetil with cyclosporine A and methotrexate as acute GVHD prophylaxis after unrelated donor allogeneic bone marrow transplantation]. AB - OBJECTIVE: To evaluate the efficacy and safety of mycophenolate mofetil (MMF) in combination with cyclosporine A (CsA) and methotrexate (MTX) for prevention of acute graft versus host disease (GVHD) after unrelated donor allogeneic bone marrow transplantation (allo-BMT). METHOD: Twelve cases of unrelated donor allo BMT were evaluated in a single center trial. The acute GVHD was prevented with 1 g MMF daily in addition to CsA 3 mg x kg(-1) x (-1) and MTX 10 - 15 mg at post BMT day1, day3, day6 and day11. RESULTS: Acute GVHD was found in one case (Grade IV) at the seventh day and two cases (Grade II) at the tenth day and seventeenth day after BMT. These patients were treated with a combination of MMF, methyprednisolone and CsA. The common adverse hematologic events of MMF was leukopenia. CONCLUSION: The preliminary study showed that MMF could be used effectively and safely for prevention of acute GVHD in unrelated donor allo-BMT. PMID- 11877054 TI - [Immunorelated pancytopenia]. AB - OBJECTIVE: A report of a group of patients with pancytopenia which might be related to abnormal immunity. METHODS: The clinical and laboratory features of 29 patients with positive bone marrow mononuclear cell (BMMNC) Coombs tests were analyzed. RESULTS: Most of these cases were pancytopenia with normal or decreased bone marrow cellularities and increased normoblasts. They were all found to have negative results of conventional hemolysis tests and hematopoietic nutrient determination and had no evidence of malignant clonal hematopoiesis. However, all of them showed positive BMMNC-Coombs test and good response to corticosteroids. CONCLUSION: Immunorelated pancytopenia might be caused by abnormal immunity (especially autoantibody) mediated destruction or abnormal function of blood cells. It is beneficial to differentiate immunorelated pancytopenia from aplastic anemia or myelodysplastic syndromes. PMID- 11877055 TI - [Relationship between soluble Fas ligand levels and complications after allogeneic bone marrow transplantation]. AB - OBJECTIVE: To evaluate the implications of soluble Fas ligand (sFasL) in acute graft-versus-host disease (aGVHD) and discriminating symptoms of aGVHD from those of infection. METHODS: Plasma levels of sFasL were assessed in 84 plasma samples from 13 patients after allogeneic BMT using a sandwich enzyme-linked immunological assay (ELISA). Plasma sFasL levels of the patients before BMT and at different time points in the post-BMT period were measured. The results were analysed for correlation with aGVHD and infections. RESULTS: Plasma sFasL levels were significantly higher in patients with grade II - IV aGVHD than that in those with grade 0 - I aGVHD (P = 0.02). There was no statistic difference in plasma sFasL levels between the infectious and non-infectious patients. In the seven grade II - IV aGVHD patients, the plasma sFasL levels pre-BMT were much lower than that in the six grade 0 - I aGVHD patients. CONCLUSIONS: sFasL may be useful for the diagnosis of aGVHD and for differentiating aGVHD from other BMT related complications. The high level of plasma sFasL pre-BMT may be of importance in decreasing the occurrence of aGVHD after BMT. PMID- 11877056 TI - [Study of telomerase activity and its related genes expression in leukemia cells]. AB - OBJECTIVE: To investigate the changes of telomerase activity and its related genes expression in leukemia cells and their roles in leukemogenesis. METHODS: The telomerase activity in bone marrow cells of 47 leukemia patients were detected by a semi-quantitative telomeric repeat amplification protocol (TRAP) silver staining, and the expression of telomerase related genes hEST2, TP1 and hTR by semi-quantitative reverse transcriptase polymerase chain reaction (RT PCR). RESULTS: The telomerase activity and expression of telomerase related genes were significantly higher in 42 leukemia patients in progress stage than that in complete remission/chronic stage and in normal/anemia cases (P < 0.01). The telomerase activity and expression of TP1, hTR gene in 14 cases of acute leukemia complete remission (AL-CR)/chronic myelogenous (CML)-chronic stage were higher than that in normal/anemia cases (P < 0.05). Low levels of telomerase activity and telomerase related genes expression could be detected in part of normal bone marrow cells. The levels of telomerase activity and telomerase related genes expression in 3 cases of anemia increased after short-term culture. CONCLUSIONS: High level of telomerase activity and its related genes expression were specific markers of malignant hematopoietic cells and their changes might be useful for observation of treatment effectiveness in leukemia. Low level of telomerase activity was associated with hematopoietic stem cells with the ability of proliferation. hEST2 gene might be a up-regulation gene of telomerase; hTR might be related to feedback regulation of telomerase. PMID- 11877057 TI - [Expressions of P-gp, mdr1, MRP and Topo II in acute leukemia patients and their correlation with prognosis]. AB - OBJECTIVE: To investigate whether P-gp, mdr1, MRP and Topo II are the prognostic factors of clinical resistance in acute leukemia (AL) patients. METHODS: The expression of P-gp was analyzed by flow cytometry using monoclonal antibody UIC2, the expressions of mdr1, MRP and Topo II by RT-PCR in 45 AL samples. RESULTS: The expressions of P-gp, mdr1 and MRP were significantly higher in resistant group than that in sensitive group (P < 0.01). The expression of Topo II was lower in resistant group, but the difference was not statistically significant. Logistic regression of univariate analysis showed that the overexpression of P-gp, mdr1 and MRP, the lower expression of Topo II and age over 55 were the prognostic factors of clinical resistance in AL. Sex, initial white blood cell count (WBC), bone marrow (BM) blast percentage and FAB subtypes were not significant for the clinical resistance. Multivariate analysis adjusted by above factors showed that the overexpression of P-gp, mdr1 and MRP, the low expression of Topo II and age over 55 were still highly significant for clinical resistance. It was also found that there were significant correlations between P-gp, mdr1, MRP and complete remission in 36 newly diagnosed AL. Significant correlations were observed between P-gp and mdr1, mdr1 and MRP, and P-gp and MRP expression in all AL cases and in clinical resistance group (P < 0.001). CONCLUSIONS: The overexpression of P-gp, mdr1 and MRP, and the lower expression of Topo II are the unfavorable prognostic factors of clinical resistance in AL. There is correlation between mdr1 and MRP expressions. PMID- 11877059 TI - [Carrier detection and prenatal diagnosis for hemophilia A]. AB - OBJECTIVE: To establish a simple, rapid carrier detection and prenatal diagnosis system for hemophilia A. METHODS: Intron 22 inversion in FVIII gene was directly examined by long distance polymerase chain reaction. Polymorphism of factor VIII intragenic RFLP of Bcl I, STR within intron 13 and 22, and extragenic DXS 52 (St 14) VNTR loci by hereditary linkage analysis were assayed. RESULTS: The diagnostic rates of these loci were 47.6% (intron 22 inversion), 27.8% (Bcl I), 28.6% and 29.4% (STR within intron 13 and 22), and 81.3% (DXS52), respectively. The overall diagnostic rate in 21 families was 94.7%. CONCLUSIONS: The diagnosis in hemophilia A patients or carriers can be made if intron 22 inversion is present. The intragenic and extragenic loci hereditary linkage analysis could be used to establish the diagnosis in intron 22 inversion negative patients. PMID- 11877058 TI - [Chromosome t (8; 21) and t (15; 17) in a patient with acute myeloid leukemia]. AB - OBJECTIVE: To report an acute myeloid leukemia (AML) patient with both chromosome t (8; 21) and t (15; 17). METHODS: Chromosome specimen was prepared by short-term culture of bone marrow cells, karyotype analyses by R-banding technique, and fusion genes by reverse transcriptase-polymerase chain reaction (RT-PCR). RESULT: Karyotype analyses showed the appearance of 45, X, -X, t (8; 21), t (15; 17), and RT-PCR assay revealed AML1/ETO and PML-RARalpha fusion gene transcripts. CONCLUSION: AML with both t (8; 21) and t (15; 17) might be recognized as a a novel subtype of acute leukemias. PMID- 11877060 TI - [Exploration of high expressions of retroviral vectors containing hFIX minigene in murine myoblast cells]. AB - OBJECTIVE: To explore the effect of different vector structure on hF IX protein expression in murine myoblasts in order to construct more efficient expression vector for my oblast-mediated gene therapy for hemophilia B. METHODS: Three types of retroviral vectors pLMe2 IX m(2)SN (forward Me2: Me2 inserted into enhancer region of 3'LTR in forward orientation), pLMe2 IX m(2)SN (reversed Me2) and pL IX m(2)SN which contained intron m(2) and two copies of MCK enhancer (Me2) were transferred into packaging cell line PA317. The hFIX expression level in selected single and mixed clones of stably transfected myoblasts were determined by ELISA and PCR. RESULTS: There was statistically significant difference in hFIX expression levels among the four vectors tested. The expression order was LMe2 IX m(2)SN (forward) > LMe2 IX m(2)SN (reversed) > L IXm(2)SN > L IX SN. The hF IX expression level of forward Me2 vector was higher than that of reversed one, because of one or two copies of reversed Me in the 5'LTR being deleted in the genome of myoblasts, but the orientation of Me or Me2 remained unchanged. CONCLUSIONS: To select a tissue-specific enhancer and optimize FIX minigene construction are effective methods for increasing the expression level. Forward Me2 vector can express hFIX more stably than the reversed one. PMID- 11877061 TI - [Study of C807T polymorphism of the platelet collagen receptor alpha2 gene coding sequence in Suzhou Han population]. AB - OBJECTIVE: To study the C807T polymorphism of the platelet collagen receptor alpha(2) gene coding sequence in Suzhou Han population. This polymorphism was associated with the expression density of integrin alpha(2)beta(1) on the platelet surface. METHODS: An amplified 1332 bp fragment of human alpha(2) gene from 110 unrelated individuals was analyzed by Bgl II and Ase I restriction assay. The resulting products were electrophoresed in 8% polyacrylamide gels and stained with ethidium bromide. RESULTS: The allelic frequencies were 0.291:0.709 for T807 allele: C807 allele in Suzhou Han population. The total theoretical heterozygous rate was 41%. The genotypic frequencies were 0.018:0.546:0.436 for homozygous 807T allele: heterozygous 807C/T allele: homozygous 807C allele. CONCLUSION: The C807T polymorphism of the platelet collagen receptor alpha2 gene in Suzhou Han population was different from that observed in other populations. PMID- 11877062 TI - [Thrombopoietic effect of recombinant human thrombopoietin gene transferred to mice mediated by electric pulse on normal and experimental thrombocytopenia mice]. AB - OBJECTIVE: To investigate the thrombopoietic effect of recombinant human thrombopoietin (rhTpo) gene transferred by electric pulse to normal and experimental thrombocytopenia mice. METHODS: Eukaryotic high expressing plasmid pcDI/Tpo was constructed by gene recombinant technology. 200 microg of the recombinant plasmid was injected into quadriceps femoris muscle of normal and experimental thrombocytopenia mice. Six times of electric pulse at 100v, 1Hz, 40ms were given immediately after the injection.The expression of rhTpo gene and its protein were assayed by RT-PCR and Western Blotting, respectively. Serum Tpo concentration was assayed by ELISA method. RESULTS: The recombinant plasmid pcDI/Tpo was successfully constructed. The expression of mRNA and protein of rhTpo gene was detected in the skeletal muscle of mice after transfection. Serum Tpo level increased from 328 +/- 89 ng/L to 1185 +/- 264 ng/L, and the platelet level of transfected mice increased from (259 +/- 27) x 10(9)/L to (640 +/- 31) x 10(9)/L. After injection with carboplatin, the platelet level decreased, but the nadir point was higher in pcDI/Tpo group than that in control group, and the recovery time of platelet count in pcDI/Tpo group shortened. CONCLUSION: The rhTpo gene could be effectively transfected to mice by electric pulse and played thrombopoietic role in vivo. PMID- 11877063 TI - [Changes in the activity of platelet L-arginine/nitric oxide pathway in hypercholesterolemia patients]. AB - OBJECTIVE: To investigate the changes of the platelet L-arginine (L-Arg)/nitric oxide (NO) pathway in hypercholesterolemia patients. METHODS: The platelet NO production in 21 hypercholesterolemia (HC) patients and 26 normal individuals was assayed by spectrum, the NOS activity and the transportation of L-Arg were determined by (3)H-labelled L-Arg. RESULTS: The NO production in hypercholesterolemic platelets [(24.06 +/- 3.70) nmol/10(8) platelets] was decreased significantly as compared with normal controls [(28.39 +/- 4.45) nmol/10(8) platelets] (P < 0.01), and so did the NOS activity [(1.46 +/- 0.47) pmol/10(8) platelets] vs (1.81 +/- 0.50) pmol/10(8) platelets]. The transportation of L-Arg by hypercholesterolemia platelets also decreased significantly than that by normal controls. The Vmax in the former [(46.84 +/- 3.39) pmol x (10(7) platelets)(-1) x min(-1)] was significantly lower than that in the latter [(53.89 +/- 3.45) pmol x (10(7) platelets)(-1) x min(-1)]. There was no difference between the two groups in K(d) (P > 0.05). CONCLUSIONS: The activity of the L-arg/NO pathway in hypercholesterolemia platelets is decreased significantly as compared with normal controls, implying that it is probably one of the reasons for the hyperactivity of hypercholesterolemia platelets. PMID- 11877064 TI - [Human cytomegalovirus inhibits the proliferation of CFU-MK in vitro]. AB - OBJECTIVE: To investigate the effect of human cytomegalovirus (HCMV) on the proliferation of colony forming unit-megakaryocyte (CFU-MK). METHODS: Semi-solid CFU-MK culture system was used to observe the effect of HCMV AD169 strain on CFU MK growth of 20 cord blood samples. HCMV DNA and immediate early antigen (IEA) mRNA in CFU-MK were detected by in situ-polymerase chain reaction (IS-PCR) and reverse transcriptase-polymerase chain reaction (RT-PCR). RESULTS: HCMV AD169 suppressed the differentiation and proliferation of CFU-MK in vitro significantly. The suppression was in a dose-dependent fashion. HCMV DNA was successfully detected in colony cells from viral infection group, and did the expression of HCMV IEA mRNA. CONCLUSION: HCMV AD169 can directly infect megakaryocyte progenitor and suppress their proliferation and differentiation. PMID- 11877065 TI - [High efficient expression of recombinant human platelet factor IV in Pichia pastoris]. AB - OBJECTIVE: To express recombinant human platelet factor 4 (PF4) in methanol yeast. METHODS: PF4 cDNA amplified by PCR was cloned into the mating factor alpha (MFalpha) signal sequence, downstream the alcohol oxidase 1 (AOX 1) promoter of the pPIC9 vector. The reconstructed vector was subsequently integrated into the methanol yeast P. pastoris strain GS115. The highly expressing strain was selected and the protein sequences and their biological activities were assayed. RESULTS: The amino acid sequence of rhPF4 was the same as that of natural one. The rhPF4 can neutralize the anticoagulation effect of heparin in a dose dependent manner. CONCLUSION: The amount of rhPF4 expressed in methanol yeast meets well the scale for manufacturing. The biological activities of the rhPF4 were the same as that of natural PF4. PMID- 11877066 TI - [Clinical significance of hemostatic molecular markers in acute leukemia]. AB - OBJECTIVE: To study the changes of hemostatic molecular markers in acute leukemia (AL) for elucidating their clinical significance. METHODS: A series of hemostatic molecular markers, including TF, TFPI, TAT, PAP, u-PA and u-PAR were measured in 82 acute leukemia. RESULTS: The plasma levels of TF, TAT, u-PA and u-PAR were elevated significantly at diagnosis in all AL patients tested, while TFPI, PAP did only in AML. After treatment, TF and TAT remained high in AML, u-PA and u-PAR were still high in un-remission patients. PAP and u-PA remarkably elevated in patients with severe hemorrhage. CONCLUSIONS: There existed hemostatic abnormality as well as hyperfibrinolysis, which varied with leukemia types and was ameliorated with clinical improvement. The measurement of TF, TAT and PAP may provide useful information for the diagnosis of DIC. u-PA and u-PAR may be considered as a useful indicator for prognosis. Patient with severe hemorrhage should be treated with antifibrinolysis drugs. The prevention of hypercoagulability after treatment should be kept in mind. PMID- 11877067 TI - [Establishment of EBV-immortalized lymphoblast cell lines from three Chinese Fanconi anemia patient and their subtyping]. AB - OBJECTIVE: To establish EBV-immortalized lymphoblast cell lines from Chinese Fanconi anemia (FA) patients. METHODS: The establishment of lymphoblast cell lines was by EBV-immortalization and subtyping of the cell lines by cell fusion, mytomycin C (MMC) inhibition analysis and/or immunoprecipitation and Western blot. RESULT: (1) Three lymphoblast cell lines were established, two of them were sensitive to MMC with a IC (50) < 10 nmol/L. (2) The subtypes of all the three patients belong to FA-A group. CONCLUSION: Complementation analysis was suitable for all FA patients with MMC-sensitive lymphoblastic cell lines, but this method is time consuming. Immunoprecipitation and Western blot are much faster for subtyping but could only be used for FA patients whose related genes had been cloned and specific antibodies been developed. PMID- 11877068 TI - [Sequential intensified immunosuppressive therapy combining with hematopoietic growth factors in the treatment of severe aplastic anemia]. AB - OBJECTIVE: To explore more effective regimen for reducing early mortality of severe aplastic anemia (SAA) and improving therapeutic effectiveness. METHODS: Antilymphocyte globulin/antithymocyte globulin (ALG/ATG) and cyclosporine A (CsA) (sequential intensified immunosuppressive therapy, SIIST), with or without hematopoietic growth factors (HGFs) were administered to 73 SAA patients in a prospective randomized clinical trial to test the effectiveness of the addition of HGFs for the patients. RESULTS: The response rate of SIIST with HGFs group was significantly higher than that of SIIST alone group (89.2% vs 63.9%), with lower rates of early infection (24.3% vs 55.3%) and mortality (4.0% vs 16.7%), shorter duration of cytopenia and blood transfusion dependence and faster recovery of bone marrow hematopoiesis. The addition of HGFs to SIIST was tolerated well in all patients. There was no difference in the treatment outcome of the two groups with GM-CSF plus Epo or G-CSF plus Epo. CONCLUSION: The use of HGFs in combination with SIIST could reduce early infection and mortality rates and, therefore, improve the response rates in SAA patients. PMID- 11877069 TI - [Umbilical cord blood transplantation (UCBT) in thalassemia children]. AB - OBJECTIVE: To evaluate the efficacy of allo-UCBT in thalassemia children. METHODS: Five patients with beta-thalassemia major (genotypes were homozygote of beta41-42, beta654 and double heterozygosities of beta41-42/beta654, beta41-42/ 28, beta654/-28, respectively) were treated by allo-UCBT. The median age was 3 year and ten month old (31/2 - 7 2/12). Four donor-recipient pairs were HLA identical and one 1 locus mismatch. The patients received a median UCB nucleated cells (NC) of 5.5 x 10(7)/kg body weight [range (4.7 - 7.7) x 10(7)/kg] and CD(34)(+)CD(38)(-) cells 2.8 x 10(5)/kg [range (0.6 - 4.5) x 10(5)/kg] and CFU-GM 1.09 x 10(5)/kg [range (0.24 - 230.00) x 10(5)/kg]. The conditioning regimen consisted ofbusalphan 16 - 20 mg/kg, cyclophosphamide 180 - 200 mg/kg, horse antithymocyte globulin (ATG) 90 mg/kg or rabbit anti-lymphocyte globulin (ALG) 25 mg/kg and melphalan of 90 mg/m(2) was added to four cases. Cyclosporine A (CsA) alone was administered in 3 cases, and a combination of CsA and methotrexate in the other 2 cases. RESULTS: Four cases were engrafted, but one rejected and recurred thalassemia state at day 60 past UCBT. One case had autologous reconstitution. The reconstitutions were found in all patients with neutrophils 0.5 x 10(9)/L at day 19 (14 - 22), platelets > 20 x 10(9)/L at day 37 (21 - 63). Two cases developed grade I and II acute GVHD respectively. None developed chronic GVHD. Three patients survived disease-freely (survival with ex thalassemia state). The median follow-up was 18 months, the probability of survival was 100%. CONCLUSION: Allo-UCBT could reconstitute hematopoiesis with low frequency and mild grade of GVHD. It is an effective treatment for thalassemia. PMID- 11877070 TI - [The combination of cyclosporin A and androgen in the treatment of chronic aplastic anemia]. AB - OBJECTIVE: To explore the therapeutic effectiveness of combination of cyclosporin A (CsA) and androgen in the treatment of chronic aplastic anemia (CAA). METHOD: Androgen alone or combined with CsA for the treatment of CAA was compared by a randomized controlled clinical trial. RESULT: The efficacy of androgen combined with CsA (87.9%) was higher than that of androgen alone (57.1%). Therapeutic effectiveness of the combination treatment between the patients with positive and negative peripheral blood mononuclear cells (PBMNCs) inhibiting normal colony formation unit-granulocyte and macrophage (CFU-GM) test showed a significant difference (P < 0.005). CONCLUSION: Androgen combined with CsA had a much better efficacy than that of androgen alone in the treatment of CAA. The patients' PBMNCs inhibiting normal CFU-GM growth test can be used as an index of the treatment outcome. The side effects of the combination therapy are low and tolerable. PMID- 11877072 TI - [Study on new techniques for genetic diagnosis of deletional alpha-thalassemia]. AB - OBJECTIVE: To develop polymerase chain reaction (PCR) techniques for diagnosis of three commonest deletional alpha-thalassemia determinants in China: - -(SA), - alpha(3.7) and - alpha(4.2). METHOD: Three groups of primers were designed and used to amplify the respective type of the deletional alpha-thalassemia under an optimized PCR condition, followed by agarose gel electrophoresis and EB stainning. RESULT: Homozygous and heterozygotes, as well as double heterozygous of the 3 commonest deletions were successfully detected and diagnosed by the developed techniques. The results were identical to that from Southern Blot analysis. Forty two cases of alpha-thalassemia were diagnosed by these techniques. CONCLUSION: The 3 PCR-based techniques established by our lab were accurate, simple, and well reproducible for gene diagnosis of the deletional alpha-thalassemia determinants. PMID- 11877071 TI - [Apoptosis of bone marrow cells aplastic anemia patients treated with cyclosporin A]. AB - OBJECTIVE: To evaluate the effects of cyclosporin A (CsA) on the apoptosis of bone marrow cells of aplastic anemia (AA) patients. METHODS: The apoptosis of and Fas antigen expression on the bone marrow mononuclear cells (MNC) in 25 AA patients and 10 normal controls were assayed by TUNEL and FACS, respectively. RESULT: The percentage of CD(34)(+) cells was decreased significantly in AA patients than in normal controls (P < 0.05), and so did the percentage of CD(34)(+)Fas(+) cells (P < 0.05). The percentage of apoptotic cells in AA patients was higher than that of controls (P < 0.01). After the patients were treated with CsA for 2 months, the percentage of apoptotic cells was decreased (P < 0.01). The percentage of CD(34)(+)Fas(+) cells was positively correlated with that of apoptotic cells in AA patients. CONCLUSION: Fas is involved in the apoptosis of CD(34)(+) cells of AA patients. CsA could reduce the percentage of apoptosis of bone marrow cells in AA patients. PMID- 11877074 TI - [A novel rat aplastic anemia model induced by 5-fluorouracil combined with busulfan]. AB - OBJECTIVE: To develop an aplastic anemia model with hematopoietic stem cell failure in rats. METHODS: Normal Wistar rats were injected i.p. with 5 fluorouracil and 5 days later p.o. by busulfan once a week for 3 times. A 5-FU alone group and a busulfan alone group were as control. The parameters tested indices were as following: incidence and mortality of aplastic anemia rats, blood picture, bone marrow nucleated cell count, CFU-GM of bone marrow cells and histology of hematopoietic organs. RESULTS: In the 5-FU plus busulfan group, the incidence of acute aplastic anemia was 82.0% (40/49 rats), the average survival time was (30.0 +/- 6.5) days. The remained 9 rats developed chronic aplastic anemia (18.0%). The total incidence of aplastic anemia was 100%, and the mortality was 98%. The blood cells count of the aplastic anemia rats was gradually reduced to a level of half the premedications and no CFU-GM was detected, and the patho-morphological examination showed that the bone marrow was replaced by fat cells, and the lymphoid tissue of spleen, thymus and lymph-nodes were remarkably atrophied without extra-medullary hematopoiesis. The control group (5-FU treated) developed a transient hematopoiesis suppression, and recovered rapidly to normal. Three of 12 rats in the busulfan treated group developed aplastic anemia. CONCLUSION: 5-FU combined with busulfan could impair the hematopoietic stem cells and resulted in an irreversible aplastic anemia in rats, making a rat aplastic anemia model with hematopoietic stem cell failure. PMID- 11877073 TI - [Improvement of combination chemotherapy tolerance of human umbilical cord blood CD(34)(+) cells transducted with double drug resistance genes by a bicistronic retroviral vector]. AB - OBJECTIVE: To explore whether human umbilical cord blood hematopoietic progenitor cells transduced with human aldehyde dehydrogenase class 3 (ALDH3) and multidrug resistance gene (MDR1) could increase resistance to 4-hydroxycyclophosphamide (4 HC) and P-glycoprotein effluxed drugs. METHODS: A bicistronic retroviral vector G1Na-ALDH3-IRES-MDR1 cDNA was constructed and transfected the packaging cell lines GP + E86 and PA317 by LipofectAMINE method, using the medium containing VCR and 4-HC for cloning selection and ping-ponging supernatant infection between ecotropic producer clone and amphotropic producer clone, cord blood CD(34)(+) cells were enriched with a high-gradient magnetic cell sorting system (MACS), and then repeatedly transfected with supernatant of retrovirus containing human ALDH3 and MDR1 cDNA under stimulation of hematopoietic growth factors. PCR, RT-PCR, Southern blot, Northern blot, FACS and MTT assay were used to evaluate the transfection and expression of the double genes. RESULTS: The purity of cord blood CD(34)(+) cells was approximately 91% and the recovery rate was 72%. The highest titer of recombinant amphotropic retrovirus in the supernatant was up to 6.5 x 10(5) CFU/ml. The efficiency of gene transduction was 18%, 20% and 16.7% tested by colony formation, PCR and FACS, respectively. Rhodamine 123 efflux showed 16% transduced cells with P-gp function. No helper virus was found by both nested PCR and rescue assay. The MTT analysis showed a 3.5 to 6.8-fold increase of resistance of transducted cells to cyclophosphamide and P-glycoprotein effluxes drug as compared with the nontransduced cells. CONCLUSION: The efficiency and co-expression of this dual genes transfer system provided a foundation for ameliorating combination chemotherapy toxicity in clinical trial. PMID- 11877075 TI - [The experimental study of transfected sFlt-Ig gene on K562 leukemia cell growth in nude mice]. AB - OBJECTIVE: To observe the influence of transfected soluble vascular endothelial growth factor (VEGF) receptor (sFlt-1) gene on K562 leukemia cell growth in vivo. METHODS: (1) The binding region of VEGF receptor (Flt-1) ligand was combined with fragment of IgH stable region to construct Flt-Ig fusion gene and insert into pcDNA3 vector. (2) By using electroporation, the pcDNA3/Flt-Ig was transfected into K562 leukemia cells, and selected by G418. Flt-Ig mRNA expression was detected by RT-PCR. (3) The transfected pcDNA3/Flt-Ig and pcDNA3-Ig K562 cells were respectively transplanted into nude mice and the tumor volume was dynamically measured. RESULTS: Five subclones of K562 cells with high expression of Flt-Ig gene have been established, one of them was transplanted into 6 nude mice. The tumor volume of experimental mice was obviously smaller than that of control mice, about one half of the control group (P < 0.05). CONCLUSION: The growth of transfected pcDNA3/Flt-Ig K562 cells was significantly inhibited. It is possible that soluble Flt-Ig protein secreted from K562/Flt-Ig cells neutralized VEGF produced from tumor cells, therefore inhibited the tumor angiogenesis. PMID- 11877076 TI - [The study of low dose radiation inducing hormesis effect on hematopoietic system]. AB - OBJECTIVE: To study the hormesis effect on hematopoietic system induced by low dose radiation. METHOD: CFU-GM and BFU-E were cultured in methylcellulose semi solid culture system, levels of GM-CSF and IL-3 were assayed by ELISA and mRNA levels of GM-CSF, G-CSF, IL-3 by in situ hybridization, narrow line hybridization and Northern blot. RESULTS: (1) The in vitro yields of CFU-GM and BFU-E from radiated mice was higher than that from the control. (2) The serum protein level of GM-CSF increased obviously than that of controls; (3) mRNA levels of GM-CSF and G-CSF were also increased. CONCLUSION: There is hormesis effect on hematopoietic system induced by low dose radiation, which may be related to the increasing of cytokines. PMID- 11877077 TI - [In vitro study of the killing activities on H-2(d) murine normal and tumor cells by Ly49A gene transfected lymphocytes]. AB - OBJECTIVE: To observe the killing activities of Ly49A gene transfected lymphocytes of C57BL/6 mice to normal and tumor cells of BALB/c mice. METHODS: pLXSN-Ly49A retrovirus vector was constructed and packaged with PA317 cell line. The lymphocytes of C57BL/6 mice were transfected by culture with virus producing PA317 cells. The Ly49A expression rate on the transfected lymphocytes was detected by flow cytometry and the killing activities of the transfected lymphocytes to normal and tumor cells of BALB/c mice were assayed by MTT method. RESULTS: The Ly49A expression rates of C57BL/6 mice lymphocytes transfected with pLXSN-Ly49A for 24 hours, of those transfected with pLXSN and nontransfected control were (46.67 +/- 0.35)%, (18.73 +/- 0.85)%, and (19.60 +/- 0.27)%, respectively. The killing activity of the transfected lymphocytes to 4T(1) tumor cells remained almost the same as that of the control (P > 0.05), but to normal fibroblasts decreased sharply (inhibiting rate 22% - 25%). CONCLUSION: The Ly49A transfected C57BL/6 mice lymphocytes could kill BALB/c mice tumor cells as effectively as the control did, but the activity decreased sharply to normal BALB/c mice cells, which would be instructive for resolving graft versus host disease after allogeneic bone marrow transplantation. PMID- 11877078 TI - [Involvement of mitochondrial membrane potential in the homoharringtonine induced apoptosis of leukemic T-cells]. AB - OBJECTIVE: Investigation of the role of mitochondrial membrane potential (MMP) in the homoharringtonine (HHT)-induced apoptosis. METHODS: Annexin V staining, flow cytometry and confocal laser scan microscopy were used to observe the relationship between Bax, cytochrome C and MMP in the HHT-induced apoptosis of leukemic T lymphocytic line Molt-3. RESULTS: The induction of apoptosis by HHT resulted in the translocation of Bax from cytosol to mitochondrial membrane and the decrease of cellular MMP, followed by the release of cytochrome C from mitochondria to cytosol. CONCLUSIONS: Changes of mitochondrial membrane potential might play a critical role in the HHT-induced apoptosis of leukemic T-cells. PMID- 11877079 TI - [Indomethacin induces apoptosis of K562 cells through activation of caspases and elevation of intracellular free calcium]. AB - OBJECTIVE: To investigate the indomethacin (IN) induced apoptosis of K562 cells: (1) the expression, activation and subcellular localization of caspase-3 and -8; (2) the change of intracellular free calcium concentration ([fCa(2+)]i) and its mechanism; and (3) whether the cell apoptosis is cyclooxygenase (cox) dependent or not. METHODS: Changes and subcellular localization of caspase-3 and -8 were observed by laser scanning confocal microscopy (LSCM); expression and activation of caspase-3 and -8 proteins by Western blotting; changes of intracellular [fCa(2+)]i by LSCM coupling with a calcium-fluorescence probe and calcium chelator EGTA blocking test; and cox inhibitor effect by MTT assay. RESULTS: (1) LSCM assay and Western blotting showed that caspase-3 and -8 localized in cytoplasm and nucleus dispersedly and spottedly and were upregulated with the increasing of IN doses. Western blotting also showed the cleavage and activation of caspase-3 and -8 during IN-induced apoptosis. (2) The increase of [fCa(2+)]i in K562 cells was parallel to the increase of IN concentration regardless of the presence or absence of EGTA. But with EGTA treatment, [fCa(2+)]i was much less than that without EGTA treatment. (3) Low dose of IN or other cox inhibitors could not exert cytotoxic effects on K562 cells whereas high dose of IN could. CONCLUSION: (1) The upregulation and activation of caspase-3 and -8 play a fundamental role in apoptosis induced by IN in K562 cells. Both cytoplasm and nucleus are locations where caspase-3 and -8 localized. (2) The increase of [fCa(2+)]i may be critical in the modulation of apoptosis; the extracellular calcium influx is the main source of the elevation of [fCa(2+)]i in K562 cells and can be blocked by the calcium chelator EGTA;the release of calcium from intracellular calcium store is also an important source of the intracellular calcium which can trigger apoptosis without the extracellular calcium influx. (3) Apoptosis of K562 cells induced by IN is cox-independent. PMID- 11877080 TI - [Study on induction of leukemic cell apoptosis by antisense oligodeoxynucleotide of human telomerase RNA]. AB - OBJECTIVE: To explore the antitumor activity of human telomerase RNA antisense oligodeoxynucleotide (As-ODN) to leukemic cells and its mechanisms. METHODS: As ODN was transfected into HL-60 cells by liposomal transfection. Telomerase activity of HL-60 cells was examined by telomeric repeat amplification protocol (TRAP)-enzyme-linked immunosorbent assay (ELISA). Apoptosis was analyzed by morphology, DNA agarose gel electrophoresis and flow cytometry. RESULTS: The telomerase activity in HL-60 cells, being transfected with 0.1 - 2.0 micromol/L of As-ODN for 1 - 6 days, varied from 1.043 +/- 0.045 to 0.063 +/- 0.011 in a dose- and time-dependent manner. The proliferation of As-ODN-transfected HL-60 cells was lower than that of control, and the cells underwent apoptosis. The Ms ODN (missense ODN)-transfected HL-60 cells didn't show these changes. CONCLUSION: As-ODN can inhibit the telomerase activity of HL-60 cells specifically and induce apoptosis. PMID- 11877081 TI - [Study on the relationship between apoptosis and reactive oxygen species of cancer cell lines induced by anticarcinogens]. AB - OBJECTIVE: To investigate the relationship between apoptosis (AP) and fluorescent intensity of reactive oxygen species (ROS) in cancer cell lines induced by anticancer agents, and the mechanisms of drug sensitivity and resistance. METHOD: K562 cells were treated respectively with each of the Vp16, ADR and DDP, the fluorescent intensity of ROS and AP percentage of K562 cells at different concentration of the drugs treated were assayed by flow cytometry (FCM). RESULT: (1) After treatment with these drugs for 24 hours, the fluorescent intensity of ROS and AP percentage of K562 cells increased obviously as compared with that of control groups. (2) The optimum concentrations of the drugs for ROS production and AP of K562 cells were determined. CONCLUSION: Anticancer agents induced AP of K562 cells, which might be through stimulating ROS production. PMID- 11877082 TI - [The mutation of anti-CD3 antibody (HIT3a) gene and its expression]. AB - OBJECTIVE: To improve the expression of anti-CD(3) single chain Fv (scFv) by site mutation and identify its biological activity. METHODS: Anti-CD(3) scFv gene was mutated by PCR, the target clones were screened by both the fingerprints of DNA restriction endonuclease digestion and Western blot, the antigen-binding activity of scFv was examined by FACS, competitive inhibition was performed with (125)I labeled HIT3a and the cytotoxic effect mediated by the anti-CD(3) scFv-activated T lymphocytes was analyzed by (51)Cr-released assays. RESULTS: The DNA sequencing showed that the 6th amino acid of the anti-CD(3) antibody (HIT3a) heavy chain gene was mutated from E (GAG) to Q (CAG). The expression of mutated anti-CD(3) scFv (m2) was increased by 100 times higher than that of the parent scFv, and there was no difference in the Jurkat cell (CD(3)(+))-binding activity between the (m2) and parent scFv. The preliminary results of competitive assays showed that m2 could partially block the sites of CD(3)(+) Jurkat cells where the parent antibody bound to. Cytotoxicity assays demonstrated that CD(3)AK cells induced by IL-2 and m2 showed stronger cytotoxic effect than that of LAK cells induced by IL 2 alone in vitro. CONCLUSION: By site mutation, a high expression fragment m2 of anti-CD(3) scFv antibody was obtained. The results of some experiments indicated that m2 could bind to CD(3)(+) Jurkat cells, furthermore, by co-stimulated with IL-2, it could activate peripheral T lymphocytes and induce CD(3)AK cytotoxic effect. PMID- 11877083 TI - [Effects of selective RAR or/and RXR retinoids on the proliferation and differentiation of NB4 cells and their mechanisms]. AB - OBJECTIVE: To investigate the effects of RARbeta selective agonist and RARalpha antagonist (RARbeta +/RARalpha -) BMS453 in combination with RXR selective agonist (RXR+) BMS649 on the proliferation and differentiation of NB4 cells, and illustrate the mechanism. METHODS: The proliferation and differentiation of NB4 cells were detected by cell count, morphological observation, NBT reduction assay, immunofluorescence analysis, flow cytometry and RT-PCR. RESULTS: BMS453 in combination with BMS649 could significantly inhibited the growth of NB4 cell in the manner of dose and time dependence. NB4 cells treated with BMS453 and BMS649 were irreversibly committed to morphologically and functionally more differentiated granulocytic cells. When NB4 cells were treated with BMS453 and BMS649 for 0, 1, 3, 12, 24 and 48 h, RARalpha, RARbeta and RXRalpha expressions were up regulated at 1 h and 3h, respectively. As compared to ATRA, the situations had no significant difference. In contrast, BMS453 or BMS649 alone was ineffective on NB4 cells. CONCLUSION: BMS453 (RARbeta+/RARalpha-) in combination with BMS649 (RXR+) significantly and synergistically inhibit proliferation of NB4 cells and induce them into granulocytic differentiation, the mechanism of which may be mediated by the AF-2 activity of RXR. PMID- 11877084 TI - [Expression of adhesion molecules on CD34(+) hematopoietic precursor cells from normal human bone marrow, cord blood and mobilized blood]. AB - OBJECTIVE: To investigate the expression of adhesion molecules on CD(34)(+) hematopoietic precursor cells from normal human bone marrow, cord blood and mobilized peripheral blood, and the mechanism of peripheral blood precursor cells mobilization. METHODS: CD(34)(+) hematopoietic cells were separated from bone marrow, cord blood and mobilized peripheral blood by CD(34) MultiSort Kit immunomagnetic bead system. The purity was examined by FACSort. The CD(34)(+) cells and post-short-term cultured CD(34)(+) cells were labeled in an indirect immuno-fluorescence procedure with adhesion molecules CD(11a), CD(18), CD(44), CD(49d), CD(54), CD(58) and CD(62L) monoclonal antibodies and assayed by FACSort. RESULTS: The expression of CD(11a), CD(18), CD(49d), CD(54), CD(58) and CD(62L) of mobilized peripheral blood CD(34)(+) cells was lower than that of bone marrow ones, especially for CD(49d) and CD(62L). Similar to mobilized peripheral blood CD(34)(+) cells, cord blood CD(34)(+) cells also showed a lower expression of CD(11a), CD(18), CD(44), CD(49d), CD(62L) than that of bone marrow ones, especially for CD(62L), but expression of CD(54) was higher than that of bone marrow and mobilized peripheral blood CD(34)(+) cells. CONCLUSION: The expressions of cell adhesion molecules on CD(34)(+) cells in normal bone marrow, cord blood and mobilized peripheral blood were quite different, the mechanism of peripheral blood precursors mobilization might be related to downregulation of cell adhesion molecule expression. PMID- 11877085 TI - [The difference of intracellular cytokines produced by different lymphocyte subsets of cord blood and adult peripheral blood]. AB - OBJECTIVE: To explore the difference of intracellular cytokines between cord blood and adult peripheral blood and elucidate its significance for GVHD. METHOD: Different lymphocyte subsets were categorized in cord and adult peripheral blood by monoclonal antibodies with 3 color fluorescence using flow cytometry. The IL 2, TNF-alpha and IFN-gamma produced by various lymphocyte subsets were compared between the two groups. RESULT: (1) In cord blood, dominant T cells were CD(4)(+)CD(45RA)(+), while the major T cells were both CD(4)(+)CD(45RO)(+) and CD(8)(+)CD(45RO)(+) in adult peripheral blood. (2) The quantities of IL-2, TNF alpha and IFN-gamma produced by cord blood lymphocytes were lower than that of adult peripheral blood lymphocytes. Moreover, the majority of cells that produced cytokines in the cord blood were CD(4)(+)CD(45RA)(+), but in adult peripheral blood were mainly CD(4)(+)CD(45RO)(+) and CD(8)(+)CD(45RO)(+). CONCLUSION: As compared with adult peripheral blood, the cord blood not only possessed different proportion of lymphocyte subsets, but also significant low level of cytokines. The differences of various lymphocyte subset proportions and lower level of cytokines in cord blood probably related to the lower incidence of GVHD in cord blood stem cell transplantation. PMID- 11877086 TI - [In vitro cotransfer human multidrug resistance gene (mdr-1) and dihydrofolate reductase gene (DHFR) into human CD(34)(+) progenitor cells to broaden the spectrum of drug resistance]. AB - OBJECTIVE: To explore the feasibility of cotransferring human mdr-1 gene and DHFR gene into human CD(34)(+) progenitor cells to broaden the spectrum of drug resistance and improve the tolerance of myelosuppression following combination chemotherapy. METHODS: The recombinant retroviral vector pSF-DIM containing mdr-1 and DHFR (L22Y) gene was constructed by introducing IRES sequence into vector FMCF which enable highly efficient gene expression in early hematopoietic cells. The retrovirus titers were raised by repeated supernatant cross infection between the amphotropic and ectropic retroviral packaging cells. Human CD(34)(+) progenitor cells were transduced by supernatant infection. Expression of P-gp was detected by flow cytometry. Integration of the foreign drug resistance gene in CD(34)(+) cells was determined by PCR. Drug resistance was evaluated by CFU-GM assay. RESULT: Integration of the two foreign drug resistance genes was detected in the CD(34)(+) cells after pSF-DIM transduction. Compared with the untransduced group, the expression of P-gp elevated by 10.98% after gene transduction and the CFU-GM yields were significantly increased at 48 nmol/L of MTX and 10 ng/ml or 12 ng/ml of taxol (P < 0.05). CONCLUSION: The retroviral vector pSF-DIM can mediate mdr-1 and DHFR gene integration and co-expression in human hematopoietic progenitor cells so as to broaden the spectrum of drug resistance. PMID- 11877087 TI - [Interferon-alpha directly inhibits thrombopoietin-induced megakaryocyte proliferation and differentiation]. AB - OBJECTIVE: To characterize the inhibitory effect of interferon-alpha (IFN-alpha) on megakaryocyte proliferation and differentiation stimulated by thrombopoietin (Tpo). METHODS: Normal murine megakaryocytes were prepared by three different cell purification procedures. Megakaryocyte development was studied by semi-solid and suspension cultures and assessed with megakaryocyte colony forming unit (CFU MK), acetylcholinesterase activity (AChE) as well as megakaryocyte count, diameter and DNA content. RESULTS: IFN-alpha prohibited CFU-MK formation and decreased AChE of megakaryocytes. The inhibitory effects were positively related to doses of IFN-alpha and could not be overcome by escalating exogenous Tpo or depriving accessory cells in bone marrow culture. IFN-alpha affected all aspects of megakaryocyte development, including decreases in megakaryocyte count, diameter and geometric mean polyploidy. CONCLUSION: IFN-alpha directly inhibits Tpo-induced megakaryocyte proliferation and differentiation in a dose-dependent manner. PMID- 11877088 TI - [In vitro autonomous growth of megakaryocytic progenitors and the role of thrombopoietin (Tpo) in essential thrombocythemia]. AB - OBJECTIVE: To investigate in vitro autonomous growth of megakaryocytic progenitors and the role of Tpo in essential thrombocythemia (ET). METHODS: Purified bone marrow and peripheral blood CD(34)(+) cells and nonadherent mononuclear cells (NMNC) from patients with ET were cultured to assay the spontaneous megakaryocyte colony formation. The autocrine and paracrine of Tpo in the spontaneous growth cells in liquid culture were examined by RT-PCR. The effect of Tpo on megakaryocytic progenitors was observed by addition of Tpo into CD(34)(+) cell culture. RESULTS: There was no spontaneous CFU-MK growth in purified CD(34)(+) cell culture and spontaneous megakaryocyte colonies disappeared at NMNC cell concentrations of 10(4)/ml and 10(3)/ml. There was no Tpo expression in the spontaneous growth cells in liquid culture. Addition of Tpo to the culture of purified CD(34)(+) cells resulted in the formation of megakaryocyte colonies. CONCLUSION: In vitro spontaneous CFU-MK formation in ET patients is not from autonomous growth of megakaryocyte progenitors. There is no autocrine or paracrine Tpo secretion. Megakaryocyte progenitors may be hypersensitive to Tpo or other MK-CSF. PMID- 11877089 TI - [Relative study of soluble syndecan-1 and prognosis of patients with multiple myeloma]. AB - OBJECTIVE: To investigate the role of soluble syndecan-1 in the pathogenesis of human multiple myeloma (MM). METHODS: Serum level of soluble syndecan-1 of patients with MM and plasma cell leukemia was determined by ELISA. RESULTS: (1) The median serum soluble syndecan-1 concentrations of MM patients and controls were 111 ng/ml (41 - 6,300 mg/L) and 63 mg/L (10 - 163 mg/L), respectively (P < 0.005). (2) For 47 myeloma patients, increased serum syndecan-1 concentrations at diagnosis were associated with a poor prognosis. Patients with serum syndecan-1 levels below or above 166 mg/L had significantly different survival times (median > 48 months or < 18 months, respectively, P < 0.0001). (3) Serum soluble syndecan 1 levels were correlated with beta-2 microglobulin concentrations and the percentage of plasma cell. CONCLUSION: Serum levels of circulating soluble syndecan-1 correlated with tumor mass and may exert pleiotropic effects on myeloma cell behavior. PMID- 11877090 TI - [Expression of recombinant human Flt3 ligand in Escherichia coli and its purification and characterization]. AB - OBJECTIVE: To establish a highly efficient expression system of recombinant human Flt3 ligand (rhFL) in E. coli and a suitable purification method of the expressed products. METHOD: Human FL encoding cDNA was introduced into pProEXHT plasmid to express a 6 x His-FL fusion protein in E. coli. The fusion protein expressed in inclusion body was isolated, solubilized and refolded, and then purified by chromatography on a metal-chelating affinity column (MCAC). Its activity was detected by stimulating the proliferation of CD(34)(+) cells. RESULT: The amount of rhFL expressed was about 15% of total bacterial proteins and the purity of rhFL was 90% after MCAC. The combination of rhFL, granulocyte colony-stimulating factor and erythropoietin could stimulate CD(34)(+) cells to a 400 fold expansion. CONCLUSION: The purified rhFL had a potent activity to stimulate hematopoietic stem cells to expanse in vitro. PMID- 11877091 TI - [Clinical study of P-gp, and bcl-2 protein expression in non-Hodgkin's lymphomas patients]. AB - OBJECTIVE: To investigate the relationship between the expression of P-gp, P26 bcl-2 and the prognosis in intermediate and high grade non-Hodgkin's lymphomas (NHL). METHODS: Sixty cases of intermediate and high grade NHL were retrospectively reviewed using immunohistochemical method. All patients were received CHOP chemotherapy over 4 courses. RESULTS: P-gp was identified in 15 and P26-bcl-2 in 25 cases. The 3-year survival rates for patients with P26-bcl-2 (+) and P26-bcl-2 (-) were 37.64% and 76.80%, respectively (P < 0.005), and for patients with both positive P-gp and P26-bcl-2 and both negative were 15.38% and 48.48%, respectively (P = 0.038). CONCLUSIONS: There is direct relationship between the P-gp, P26-bcl-2 protein expression and the prognosis in intermediate and high grade non-Hodgkin's lymphoma. PMID- 11877092 TI - [The Immunophenotypical features of t (8; 21) (q22; q22) acute myeloid leukemia]. AB - OBJECTIVE: To study the predictive value of immunophenotypical features in t (8; 21) (q22; q22) acute myeloid leukemia (AML). METHODS: Morphological/cytochemical, flow cytometric immunophenotyping, cytogenetic analyses (MIC) and RT-PCR were performed in 294 previously untreated AML. RESULTS: (1) In 294 AML patients, t (8; 21) AML were 21.8% (64); in AML-M(2), t (8; 21) AML were 54.7%; and in t (8; 21) AML, AML-M(2) were 81.3%. (2) Compared with control group, CD(19) and CD(34) expressions were higher, and CD(33) expression was lower (P < 0.001) in t (8; 21) AML. (3) If the cut-off value of CD(19) positive was >or= 20%, CD(19) positive rate was 13.6% (40/294) in AML, and 50% (32/64) and 3.5% (8/230) (P < 0.001) in t (8; 21) AML and control group. (4) CD(19)(+) and/or CD(34)(+) t (8; 21) AML accounted for 90.6% (58/64) of t (8; 21) AML and CD(19)(-)/CD(34)(-) for 9.6% (6/64). CONCLUSION: In t (8; 21) AML, especially M(2)/t (8; 21), CD(19) and CD(34) expressions were high. CD(19) was one of predictive markers of t (8; 21) AML. PMID- 11877093 TI - [Influence of chemotherapy on hematopoietic microenvironment and effect of autologous bone marrow stromal cell infusion on the recovery of hematopoiesis after chemotherapy]. AB - OBJECTIVE: To observe the influence of chemotherapy on hematopoietic progenitor cells (HPC) and hematopoietic microenvironment (HME). To observe whether intravenous infusion of autologous bone marrow stromal cells (ABMSC) expanded in vitro can improve the hematopoietic function. METHODS: Cultures of CFU-GM, BFU-E, CFU-E and stromal progenitor cells (CFU-F) from normal control and chemotherapeutic patients were performed. The stromal function was analyzed by the assessment of the area of flask surface covered by stromal cells (ASSC) and the time when stromal cells reach confluence (TC). The recovery of hematopoietic function in short term chemotherapy group and long term chemotherapy, with or without ABMSC infusion (1.1 - 8.7) x 10(8) post chemotherapy groups was observed. RESULTS: The yields of CFU-GM, BFU-E, CFU-E and CFU-F in long term chemotherapy group were significantly lower than that in normal group or in short term chemotherapy group. There was no significant difference among three groups in the ASSC and TC. In long term chemotherapy group, the yields of CFU-GM, CFU-E, BFU-E and CFU-F after chemotherapy with BMSC infusion were significantly higher than that without BMSC infusion. In long term chemotherapy group, the lowest value of white blood cell (WBC) and platelet after chemotherapy with BMSC infusion was significantly higher than that without BMSC infusion. The times for WBC and platelets recovered to normal were significantly shorter in BMSC group than in without BMSC. No adverse reaction was observed with ABMSC infusion. CONCLUSION: Long term chemotherapy results in severe impairment in HPC and mesenchymal progenitor cell (MPC), but has no obvious influence on the in vitro BMSC confluent layer formation. Intravenous infusion of expanded ABMSC can accelerate the recovery of hematopoiesis after chemotherapy. PMID- 11877094 TI - [Risk of leukemia among medical X-ray workers in China between 1950 and 1995]. AB - OBJECTIVE: To explore the relationship between low dose radiation and leukemia and assess resultant risk of leukemia. METHODS: The leukemia incidences (1950 - 1995) in 27,011 medical diagnostic X-ray workers and in 25 782 non-radiation workers in hospital employed between 1950 and 1980 in China were compared by O/E system. RESULTS: The relative risk (RR) of leukemia in the medical X-ray workers was 2.17 (95% CI = 1.58 - 2.91, P < 0.01). The highest RR of leukemia was found in those who engaged in X-ray work before 1960 or at age under 20 years. CONCLUSION: There was a relationship between incidence of leukemia and X-ray radiation in X-ray workers and accumulation of radiation dosage and employment at younger age increased the risk of leukemia. PMID- 11877095 TI - [Aminopeptidase inhibitor Bestatin induces HL-60 cell apoptosis through activating caspase 3]. AB - OBJECTIVE: To study the variation and significance of caspase 3 activity in the process of amino-peptidase inhibitor--bestatin (BS) inducing human leukemic cell apoptosis. METHODS: Cell apoptosis was evaluated by light microscopy, TUNEL labeling and flow cytometry (FCM). Caspase 3 activity was detected by colorimetry. The mitochondrial transmembrane potentials (DeltaPsi(m)) were detected by Rhodamine123 staining. RESULTS: The apoptotic morphology, apoptotic peak on FCM and positive Annexin V(FITC) on cell membrane showed that BS could induce HL-60 cell apoptosis in a dose- and time-dependent manner. Caspase 3 activity was significantly higher in the apoptotic cells than in control cells. The apoptosis induced by BS was inhibited by AC-DEVD-CHO. The DeltaPsi(m) of cells treated with BS declined. CONCLUSION: BS induces apoptosis of human acute leukemic cells through activation of caspase 3. PMID- 11877096 TI - [Study on the transformation from myelodysplastic syndromes into acute leukemias]. AB - OBJECTIVE: To study the patterns of transformation from myelodysplastic syndromes (MDS) into acute leukemias (AL). METHODS: Leukemic transformation of MDS patients was dynamically followed up and the clinical manifestations, peripheral blood and bone marrow pictures, karyotypes, immunophenotypes, response to treatment and prognosis of post MDS acute leukemia (postMDS-AL) were observed. RESULTS: During the past eight year and seven months, 21 (13.91%) of 151 MDS patients progressed to overt leukemia with a median interval of 5 (1 - 21) months. There were no significant differences among the rates of leukemia from RA, RAEB and RAEB-t groups. The transformation was developed either gradually or rapidly. There were five parameters related to the leukemic transformation: under 40 years of age, pancytopenia, more than 0.15 blasts in bone marrow, at least two types of abnormal karyotype and combined chemotherapy. All of the 21 post MDS-AL were acute myeloid leukemia (AML); and most of them were M(2), M(4) and M(5). Two (9.52%) post MDS-AML developed extramedullary infiltration. Leukopenia was found in 47.62% of patients. Two third of the patients, whose bone marrows were generally hypercellular, showed neutropenias. After evolving into AML, 8 (47.06%) patients developed abnormal karyotypes. High expression of immature myeloid antigens, including CD(33) (49.83 +/- 24.50)%, CD(13) (36.38 +/- 33.84)%, monocytic antigen CD(14) (38.50 +/- 24.60)%, and stem cell marker CD(34) (34.67 +/- 30.59)% were found on bone marrow mononuclear cells of post MDS-AML cases. In some cases, lymphoid antigens, such as CD(5), CD(7), CD(9) and CD(19) were coexisted with myeloid antigens. A low complete remission rate (31.25%) and short survival duration with median survival of 6 (1 - 28) months were found in patients with post MDS-AML treated by induction therapy. CONCLUSION: MDS was at high risk of evolving into AML, either gradually or rapidly. Patients with post MDS-AML had specific biologic features and worse prognoses. PMID- 11877097 TI - [Effect of inhibition of telomerase activity on cisplatin-induced apoptosis in K562 cells]. AB - OBJECTIVE: To explore the effect of human telomerase reverse transcriptase (hTERT) gene antisense oligodeoxynucleotide (ASODN) on telomerase activity and cisplatin-induced apoptosis in K562 cells. METHODS: Telomerase activity was analyzed by using polymerase chain reaction enzyme-linked immunoassay (PCR ELISA), expression levels of hTERT protein by immunofluorescence and apoptosis by DNA gel electrophoresis and flow cytometry. RESULTS: There was a marked decrease in both the percentage of hTERT protein positive cells and the telomerase activity in ASODN treated cell as compared with that in control and sense oligodeoxynucleotide (SODN)-treated cells; but no difference between the latter two groups. Agarose gel electrophoresis of genomic DNA from K562 cells treated with ASODN and cisplatin combination for 72 h showed typical DNA ladder; neither did DNA from K562 cells treated with SODN plus cisplatin nor cisplatin alone. Apoptosis rates of K562 cells treated with ASODN for 24 h and then with cisplatin for 72 h were significantly increased. There was statistically significant difference in the percentage of apoptotic cells between hTERT ASODN plus cisplatin group and SODN plus cisplatin or cisplatin alone group. CONCLUSION: The hTERT gene ASODN could inhibit telomerase activity of K562 cells whereby it enhanced cisplatin-induced apoptosis of the cells. PMID- 11877098 TI - [Study of specific targeting cytotoxicity mediated by anti-CD(3)/anti-CD(20) Diabody]. AB - OBJECTIVE: To study the specific targeting cytotoxicity mediated by an anti CD(3)/anti-CD(20) diabody. METHODS: The diabody was purified by affinity chromatography and identified by Western blot assay, size exclusion chromatography, FACS and rosetting assay; the effect of the anti-CD(3)/anti CD(20) diabody mediated lysis of CD(20)-expressing tumor cells was assayed by (51)Cr release assay in vitro and by human B cell tumor nude mice xenograft model in vivo. RESULTS: The anti-CD(3)/anti-CD(20) diabody could bind both Jurkat cells (CD(3)(+)) and Daudi cells (CD(20)(+)) and appeared to be potent in targeting activated peripheral blood mononuclear cell (PBMC) to lyse Daudi cells in vitro. Furthermore, it inhibited tumor growth and prolonged the survival of mice bearing xenografted Raji cells. CONCLUSION: The diabody proved to be a potent agent for targeting peripheral blood lymphocytes to lyse CD(20) antigen expressing tumor cells in vitro and in vivo. PMID- 11877099 TI - [Affect of As(2)O(3) on the sister chromatid exchanges in human blood lymphocytes]. AB - OBJECTIVE: To assess the potential of As(2)O(3) in inducing malignant transformation at its therapeutic concentrations. METHODS: The sister chromatid exchange (SCE) in human peripheral blood lymphocytes (PBL) treated with different concentrations of As(2)O(3) was detected by the staining technique for sister chromatid differentiation. RESULTS: At 2 x 10(-6)mol/L and 1 x 10(-6) mol/L of As(2)O(3), the SCE frequency in PBLs was significantly increased as compared with the control (P < 0.01). Most of the lymphocytes were arrested at the first cell cycle (M1) and the mitotic index was significantly decreased in a dose dependent manner. At 1 x 10(-7) mol/L or less of As(2)O(3), the SCE frequency, the cell proliferation kinetics and the mitotic index in PBLs were similar to the control (P > 0.05). CONCLUSION: It seemed that As(2)O(3) might induce transformation and affect the function of lymphocytes when used within the therapeutical concentration range. PMID- 11877100 TI - [Invasive rhinocerebral aspergillosis occurred during myelosuppressive phase after chemotherapy: a case report and literature review]. AB - OBJECTIVE: To report a case of invasive rhinocerebral aspergillosis (IRA) during myelosuppressive phase after chemotherapy and the treatment outcome. METHODS: A patient with acute non-lymphoblastic leukemia (M(2a)) occurred IRA during myelosuppressive phase after chemotherapy and was treated with amphotericin B (AmB, including AmB-L), garlicin, 5-flucytosine (5-FC), fluconazole, itraconazole, ketoconazole, econazole and griseofulvin with twice of cerebrotomy lesion resections and cerebral decompressions. RESULTS: Though the survival time of this patient is a little longer than the others, she finally died of uncontrolled infection. CONCLUSION: For most of the IRA, there remains no effective treatment. The best treatment is AmB in combination with surgical excision. PMID- 11877101 TI - [Autologous peripheral blood selected CD34+ cell transplantation in the treatment of multiple myeloma]. AB - OBJECTIVE: To investigate the clinical impact of CD(34)(+) cell selected autologous peripheral blood stem cell transplantation (APBSCT) in multiple myeloma (MM). METHODS: The hematopoietic recovery time, treatment efficacy, survival rate, transplantation related morbidity and the cost were compared between 21 MM patients receiving CD(34)(+) APBSCT and 21 MM patients receiving unselected APBSCT. The age and beta(2) microglobulin level at diagnosis and disease status at the time of transplantation were similar in the two groups. The induction treatment and conditioning regimen were the same. RESULTS: Significantly lower CD(34)(+) cells were infused in the selected group as compared with the control group: 2.2 (0.5 - 14.3) x 10(6)/kg vs 9.4 (1.1 - 15.0) x 10(6)/kg, (P < 0.001). The median times to neutrophil >or= 0.5 x 10(9)/L and platelet >or= 20 x 10(9)/L were 10 and 9 days for the selected group, and 9.5 (P = 0.357) and 4.5 days (P = 0.005) for the control group, respectively. Response rates were similar in both groups (85.7% for the selected group vs 90.4% for the control group). Three year disease-free survival (32% for the selected group vs 39% for the control group) and overall survival rates (85% for the selected group vs 79% for the control group) were not significantly different. Furthermore, unselected APBSCT could reduce the cost for the transplantation. CONCLUSION: It shows that CD(34)(+) cell selected autologous transplantation is more expensive and does not improve the clinical outcome of patients with MM. PMID- 11877102 TI - [The diagnostic value of platelet glycoprotein-specific autoantibody detection in autoimmune thrombocytopenic purpura]. AB - OBJECTIVE: To evaluate the clinical significance of platelet glycoprotein specific autoantibody detection and platelet-associated IgG (PAIgG) measurement in the differential diagnosis between autoimmune thrombocytopenia and nonimmune thrombocytopenias. METHODS: Anti- GP II b/III a and anti-GP I b/IX autoantibodies were detected by a modified monoclonal antibody immobilization of platelet antigens assay (modified MAIPA), and PAIgG by competitive ELISA. RESULTS: Comparing with the measurement of PAIgG, the GP-specific autoantibody assay was less sensitive but much more specific for the diagnosis of autoimmune thrombocytopenia. CONCLUSION: The platelet GP-specific autoantibody detection is considerably better than PAIgG measurement in discriminating immune from nonimmune thrombocytopenia. PMID- 11877103 TI - [Introduction of ancient Bencao works stored overseas yet not included in Quanguo Zhongyi Tushu Lianhe Mulu (National Coalition Catalogue of TCM Works)]. AB - This paper briefly introduces 14 ancient Bencao works stored overseas yet not included in Quanguo Zhongyi Tushu Lianhe Mulu, with the information, including the authors, accomplish date, number of volume, categories, quantity of drugs, characteristics of content, store sites, and the condition of returned duplicated edition in China, and etc. Among the 14 works, 2 were accomplished in the Yuan dynasty, 9 in the Ming dynasty and 3 in the Qing dynasty. PMID- 11877104 TI - [Investigation on relation between Yang Shangshan and the classified compilation of Tai su (Comprehensive Notes)]. AB - Taisu, a book complied by Yang Shangshan by imperial decree is the earliest complete commentary and classified complation of Neijing (The Yellow Emperor's Canon of Internal Medicine) now extant. Recently, some scholars argued about the classified compilation of Taisu performed by Yang Shangshan. However, by analyzing and researching into the text and commentary of Taisu, conclusion can be drawn that the classified compilation of Taisu was indeed complied by Yang Shangshan himself. First, "youben" (certain edition) and "yiben" (an edition) in the commentary are not referring to the other editions of Taisu. Second, quotations cited from Su wen (Plain Questions) and Jiu juan (Nine Volumes) in the commentary are not used to collate Taisu. Thire, analysis made on the compilation and commentary by Yang Shangshan. Fourth, analyzing "taisu jing lun (Classical Discourses of Taisu)", the commentary of Taisu Shui lun (On water in Comprehensive Notes). PMID- 11877105 TI - [Yi lin cuo yao (Essentials of medical works) and its historical materials of medical exchanges between China and Korea]. AB - Yi lin cuo yao, including 13 chapters and 128 categories, was written by Zheng Zhengxian and collated by Yang Lishou, both Korean imperial physicians in the 16 - 17th centuries. With notation of a number of the Chinese and Korean medical books and materials in the first decade of the Li dynasty, this book is a comprehensive prescription work, very useful to clinical application. Especially with several points of the formulas from Zhong chao zhi wen fang (Sino -- Korean Prescriptions of Questioning) and zhong chao chuan xi fang (Sino -- Korean Prescriptions of Questioning) recorded in this book connected with Ren zhai zhi zhi fang lun (Renzhai's Comprehensive Prescriptions) in the Song dynasty. This book is an important record of the exchanges on medical ideas between China and Korea. However, it has nothing to do with Yi xue yi wen (Questions on Medicine) by Fu Maoguang, which records the questions and answers between Chinese and Korean officials. PMID- 11877106 TI - [Analytical study on characteristics of medical thinking in the Sui and Tang dynasties]. AB - The cultural environment of the Sui and Tang dynasties is rather loose, and medical thinking is especially active. The medical thinking of this period has the following characteristics: the thinking of compiling medical books influenced by the idea of Universe Harmony; taking the thinking of stressing human life and health; epistemology of paying attention to practice and perception; methodology with the tendency of analytical and integrative methods; ambivalent viewpoint in ghosts and gods, and destiny. PMID- 11877107 TI - [Preliminary study on the methods of systematizing the data of ancient epidemic situations]. AB - Taking the data of ancient epidemic situations in the south of the Five Ridges, as an example, the systematization materials of ancient epidemic situation guided by epidemiological methods are explored by defining the material of epidemic situation, selected compilation of relevant factors, decision of the severity, location, and its occasion etc. It is attempted to solve some problems in modernization and datalization of the collation of ancient literatures of Traditional Chinese Medicine. PMID- 11877108 TI - [Dialectic research on "shu pu" (referring a symptom) and "shu pu" (referring a place)]. AB - The name of "shu pu" come from Shen shi fang (Prescriptions of Master Shen), which was quoted by Wai tai mi yao (Medical Secrets of an Official), compiled by Wang Tao of the Tang dynasty. In the successive processes of Zheng lei ben cao (Classified Materia Medica) and Ben cao gang mu (Compendium of Materia Medica), it was mixed up with "shu pu", a symptom, in Su wen ci jin lun (On Banning of Acupuncture in Plain Questions), and "shu pu", a place, in Zhen jiu jia yi jing (A -- B Classin of Acupuncture and Moxibustion). In fact, "shu pu" was the meat of rats with no processing, which can be used externally or administered orally to treat injury due to metallic tools and abscess. PMID- 11877109 TI - [Comparison the medical prescriptions between of Zhongjing and Xiao pin fang (Jotting Prescriptions)]. AB - Based on the comparison between the prescriptions of Zhongjing's Prescriptions and Jotting Prescriptions, it can be seen that Xiao pin fang (Jotting Prescriptions) in many aspects, reflecting partly the tendency of changes in prescriptions from the Eastern Han dynasty to Liu Song period of the Northern and Southern dynasties. PMID- 11877110 TI - [History of nephritis]. AB - The nephritis is one of the common and detrimental diseases. This paper reviews the history of knowledge of nephritis, ancient and modern, and western and Chinese medicine, demonstrating that the continuous development of treatment of TCM and western medicine are all effective measures to combat nephritis. PMID- 11877111 TI - [Modern western medical education in Fujian]. AB - The modem western medical education in Fujian mainly stemmed from three aspects: medical schools established by Missionary, western schools managed by government and western medical education in the Soviet District of Western Fujian. These three educational modalities promoted the development of modern western medical education in Fujian. PMID- 11877112 TI - [A brief history of medical relief law for natural calamity in modern China]. AB - The history of medical relief law for natural calamity in China, this article shows the developmental tendency of medical relief for natural calamity, and the countermeasures are also elucidated. PMID- 11877113 TI - [The germination and preliminary formation of science of TCM formula in the pre Qin dynasty]. AB - Shennong tasted hundreds of grasses and found herbs during the legend period. The appearance of prescription should be so early if single drug was regarded as single -- item recipe. However, the explicit records in literature appeared at the end of the Spring and Autumn period and the beginning of the Warring States at the latest, evidenced by the custom of "presenting a recipe" of the Qi states. The medical books of Mawangdui, mainly Wu shi er bing fang (Prescriptions for 52 Kinds of Diseases), recorded the classification of prescriptions based on the sort of diseases, composing, preparation, method of decocting medicinal herbs, contraindication of prescriptions, and the therapeutic methods, compatibility of complex prescriptions and determination of treatment based on the differentiation of syndromes (Zheng), etc. reflected in the recipes, which could represent the primary level of science of TCM formula of the Warring States in China. The classic medical book Nei jing (Internal Canon of Medicine), including denominations, composition, principles of compatibility and rules of clinical application of concrete prescriptions, especially therapies and theories of composed prescription, have great actions on the development of science of TCM formula. PMID- 11877114 TI - [The characteristics of official and personal formulary in the Song dynasty]. AB - The development of prescription books reached its peak in the Song dynasty. There were two kinds of the book: official and personal. The former was characterized by its abundant contents, authentication and rich set -- prescriptions, while the latter was characterized by paying attention to practices, conscience and careful classification in different special areas. The two types of formulary embody the sense of splendor stage in the Song dynasty from various viewpoints. So it is useful to the development of modern pharmacology of TCM by studying the two types of formularies. PMID- 11877116 TI - [Preliminary study on the time of first appearance of "lanhousha" (scarlet fever)]. AB - There are controversies about the problem of the source of "lanhousha" (scarlet fever) in the field of modern medical history, whether it is an epidemic appeared from the ancient time in our country or imported from other countries. Now, almost all researches were carried out around the classic medical books, especially "Medical Records of Ye Tianshi", and hardly dealing it in a historical context at that time. Through contrastive analysis between the records in medical books and historical facts, it is concluded that Ye Tianshi's records of scarlet fever have nothing to do with Ye Tianshi himself, and is by author of allonym. It perhaps was exogenous cold diseases spread when a severe epidemic diseases happened at Southern Jiangsu in the 11th year of Yongzheng reign. Even though there was some scarlet fever, it was not the first spread of scarlet fever in fact. Actually, in late Kangxi reign, scarlet fever was regarded as an epidemic disease seldom seen at Southern Jiangsu. PMID- 11877115 TI - [New evidences for adapting Qian jin yao fang (Thousand golden essential prescriptions) by Office of Revising Medical Books of the Song dynasty]. AB - Qian jin yao fang (Thousand Golden Essential Prescriptions) written by Sun Simiao in the Tang dynasty has two kinds of extant editions in circulation: one is the revised edition, Bei ji qian jin yao fang (Essential Prescriptions Worth a Thousand Gold for Emergencies) revised by the Office of Revising Medical Books of the Song dynasty, the other is the Song edition, Sun zhenren qian jin fang (Master Sun's Thousand Golden Prescription) (20 volumes extant) not revised by Song people. There are great discrepancies between these two editions not only in the order of chapter contents, including the order of items in the same chapter, but also in its texts. Concerning the causes of this difference, we already made preliminary analysis in our former paper. However, there are new evidences to further explain it with our recent research. PMID- 11877117 TI - [Introduction of health administrations at the end of the Qing dynasty]. AB - During the new policy and reform of constitutionality from 1901 to 1911 at the end of the Qing dynasty, there were the contents of health administrations. A special independent organization was affiliated to the Ministries of Police and Civil Administration. The central health administration refers to the Health Section of Police, which was promoted to a Department after the Police Ministry changed to Civil Administration, including three sections: health, quarantine and arts of necromancy, astrology, and medicine. The duties of these three sections included checking the establishment of medical schools, testing doctors, administrating cleaning streets and epidemic prevention, examining and approving health rules. A Health Department was set up by the police headquarters of inner and external city in the capital, and health section was set up by police of each province. There was a police head in government of county to charge the health works. It was the first health administration in the bureaucracy system in China, and the beginning of learning from the west and modernization on health administration. PMID- 11877118 TI - [General situations of medical rescue at Shanghai public concession in the 30s of 20th century]. AB - In the 30s of 20th century, Shanghai public concession is the largest concession of old China. Health Department of Work Bureau and Fire Department administrated the rescue works in concession. The affiliated first -- aid sections and hospitals were responsible for concrete rescue works of patients. The health workers were hardworking for the health, body and mind, of all kinds of people, inside and outside, and achieved clear results. PMID- 11877119 TI - [The study of the foundation of the health organization of the Chinese People's Liberation Army (PLA)]. AB - The PLA's health work was gradually founded and developed from 1927 when the army was established. General Medical Department of the Central Military Commission, which was the leading department of health work of the PLA, was established in 1931. Then, all kinds of health organizations were established and strengthened, and uniform leader system was formed. At the same time, health workers were trained and the laws were set up. The initiatory stage was accomplished in 1933. The primary founder of the PLA's health work was He Cheng. PMID- 11877120 TI - [History of development in the transplantation of hematopoietic stem cell]. AB - Some malignant and nonmalignant hematological diseases can be cured by the transplantation of allogeneic stem cell. It is thought to be one of the landmarks in the development of modern medicine. Hematopoietic stem cell transplantation has been performed for about one hundred years in the phases of germination, early investigation and development. The great challenge now is how to perfect the technique and to extend the applied areas. PMID- 11877121 TI - [Primary study of Gu Dingfang, an imperial doctor of the Ming dynasty and his funerary objects]. AB - Through researching textually the name, alias, life story, family tree, and medical thought, medical activities and unearthed culture relics etc. of an imperial doctor, Gu Dingfang, his remarkable contribution to the reservation of ancient classic medical editions and the reality of a new creative theory on psychological therapy put forward by him are shown. His outstanding achievements not only complement the blank of medical history of the Ming dynasty, but also occupy a position in the world's medical history. PMID- 11877123 TI - [Liang Longzhang, the famous neoteric physician of the south of the Five Ridges]. AB - Liang Longzhang, a famous neoteric physician in Guangdong province, wrote Bian zheng qiu zhen (Seeking Truth from Differentiation of Symptoms and Signs) and studied medicine carefully and thoroughly. He once was employed by a British company and often went to Southeast Asia to treat the illness of royal members, so he was famous both in China and abroad. He was active in medical charitable work in Guangdong and Hongkong, and tried his best to help the poor. In the overflowed plague of Guangdong, he created "Yi shu yun qi fang lun" (Theory of Recipes Based on Mathematics of Books of Changes and the Circuit Phases and Six Climatic Factors) to save the patients and achieved remarkable effects. In addition, he appealed the government to reform after comparing the medical system of Chinese and western, and was a physician with advanced idea. PMID- 11877122 TI - [Investigation on the life of Wu Jutong, the famous physician of seasonal warm diseases, and his books]. AB - The name of Wu Jutong, a physician of seasonal warm diseases in the Qing dynasty was Tang, another name being Peiheng, and Jutong was his alias. He was born in 1758 and died in 1836 and his native place was now Huai'an city, Jiangsu province. Wen bing tiao bian (Treatise on Differentiation and Treatment of Seasonal Warm Diseases) was not finished in 1798, but 1813, after a hard efforts of 15 years. He compiled a Yi yi bing shu (A book Checking the Defects of Medical Professionals) in 1931 and arranged his medical records as books in 1833 to help latercomers. He not only researched medicine carefully and thoroughly and was brave in innovation, but also was an honest man with high morality. He opposed factions when studying, and had an open mind, collected widely from various schools, and had right attitude towards his own achievements. He was a model, and worthy to study and yearn. PMID- 11877124 TI - [Contribution of Soranus of Ephesus on Apgar score]. AB - Soranus of Ephesus was a famous gynecological physician in Roman times and one of the pioneers of gynecology, pediatrics and so on. The contribution of Soranus on Apgar score was researched by comparing Soranus critical and Apgar score of evaluation system of newborn infant. PMID- 11877125 TI - [Literature researches of Herba Lagopsis]. AB - The usage of Herba Lagopsis as a drug has a long history, and it was arranged under the item of Semen Leonuri in the herb books of past dynasties and used as a kind of Semen Leonuri. The alias of Semen Leonuri is Spica Prunellae in records, and the withered Semen Leonuri after the summer solstice is also Herba Lagopsis. Now, there are many Flora and local Flora and records of materia medica embodying it, and its function is similar with motherwort. PMID- 11877126 TI - [A review on nasopharyngeal carcinoma in ancient Chinese literature]. AB - Based on the clinical characteristics of nasopharyngeal carcinoma (NPC), a receptive standard was set up to screen related information with NPC from ancient literature of traditional Chinese medicine, while a rejection standard was also set up to prevent associated data in these literature with diseases such as lymphatic tuberculosis and lymphoma in the neck etc. Furthermore, a review was made on the etiology, pathogenesis and procedures used in the prevention and treatment of NPC recorded in ancient literature, according to the evaluation as above. The data showed that several hypotheses were present in ancient literature about etiology and pathogenesis of NPC, including healthy qi deficiency, cold in channels, or liver qi stagnation, pyrogenous toxin, phlegm coagulation and blood stasis etc. Therefore, prevention was emphasized and the main therapeutic and preventive methods for NPC were regulating emotions, nourishing qi and harmonizing ying (nutrient). PMID- 11877127 TI - [Jivaka's recipes in Indian ancient medical texts]. AB - Jivaka was very important in the history of Indian Buddhist medicine, and Indian ayurvedic classics yet ascribed some recipes to him. Jivaka's four recipes from Navanitaka, Carkadatta, and Bodhisattva - garbhastha - sutra etc. were handed down, reflecting his medical achievements. PMID- 11877128 TI - [The annotated research on "Xun du", "Xun li" as seen in excavation of the remains of Western Han dynasty]. AB - The "Collected Notes of Armed Vehicles of 4th year of Wuku Yong shi" in Han bamboo slips excavated from the remains of Han tomb in Lianyungang, Jiangsu, carries a "Xun du", while a "Xun li" is carried in the Han record paper excavated in the remains of Xuanquan of Han, Dunhuang, Gansu. Both should be "frankincense", being the earliest record of this material, demonstrating that this medicine had been imported at least in early Western Han dynasty. Thus, the record of frankincense should be 300 years ahead of the existing record. PMID- 11877129 TI - [A new interpretation for "Knockout Drops" (menghan)]. AB - Knockout drops ("menghanyao" in Chinese) is named as one of riddles in the cultural history of China. Why its is named as "menghan" has been interpreted from the viewpoint of phonology by many former solons. A new opinion is put forward from the viewpoint of pharmacology and physiology in this paper, in which a historical fact is concerned that mandala is the essential component in knockout drug. The function of sweat gland is inhibited with sweat retained inside the human body after knockout drops are taken. "Menghan" just describes the above physiological phenomenon. PMID- 11877130 TI - [Discovery and brief developmental history of electro -- convulsive therapy in mental diseases]. AB - With its introduction in 1938, electro -- convulsive therapy (ECT) rapidly became accepted as a mainstream treatment. Its usage opened a new era treatment of mental diseases. ECT is a very effective treatment for major psychosis with high incidence such as endogenous depression, catatonia in schizophrenia and so on. Until recently it still has a place in psychiatric disorders even when effective antipsychotic chemicals are applied today. PMID- 11877131 TI - [A century of developmental history of laparoscopic surgery]. AB - Till the year 2001, the development of laparoscopic surgery will have gone through a total of one hundred years. Laparoscopic surgery has made a great progress during the last more than a decade. In retrospect its history, the development of laparoscopic surgery underwent three main eras, viz. diagnostic laparoscopy, operative laparoscopy and the modern laparoscopy era. Although the laparoscopic technology started rather late in China, it develops very quickly. Along with the development of high -- tech, laparoscopic surgery will have a promising future. PMID- 11877132 TI - [History of mediastinoscopy]. AB - As an important measure for the diagnosis and treatment of mediastinal diseases, mediastinoscopy has a history of over half a century, undergoing 3 stages in its course of development, namely, exploration, mature and improvement. In China, due to historical restrain, though this technique started quite early, yet it lags far behind the advanced level of the world now. PMID- 11877133 TI - [Research on achievement on pulsology in Yi bian (Medical bank stone) by He Mengyao]. AB - Yi bian (Medical Bank Stone) was the representative works of He Mengyao, a doctor of the Qing dynasty. There were detailed introduction about pulse in the fifth volume, pulse feeling, including its sites, figures, movements, paused conditions, corresponding with four seasons and five organs, stomach - qi of pulse and pulse conditions referring to various diseases with examples and reasons. Not only was it written in an easy and fluent style reasonably and colorfully, but also the words were arranged clearly and beautifully. PMID- 11877134 TI - [Sphygmological characteristics in San zhi chan (Buddhist Zen of Three Fingers)]. AB - Zhou Xueting knew very well about the origin of sphygmology. In his works San zhi chan (Buddhist Zen of Three Fingers), he used antitheses to point out the mild pulse, which is the pulse condition of a normal people. By comparing 26 types of abnormal pulse, the outline is compact and catchy. He advocated the sense by insight, diagnosing the illness by pulse taking of three fingers. For common diseases, he adopted the method that closely combines the pulse, the symptom and the treatment together. This book occupied an important position in the studying of sphygmology. PMID- 11877135 TI - [Zhu Xi's influence on development of ancient traditional Chinese medicine]. AB - As an ancient famous philosopher and a figure of encyclopedic pattern, Zhi Xi was conversant with traditional Chinese medicine (TCM) and the art of healthcare in addition to the knowledge of philosophical knowledge and its implication, with special understanding on medical classics and its recipes, pulse taking and drug application. He exerted significant influence on the development of ancient medicine, especially in methodology which can be summarized into 3 points. 1. Advocating seeking truth by suspicious attitude; 2. Emphasizing the combined use of simplicity and erudition and from erudition to simplicity; 3. Nature and natural law, manifesting rather high rational dialectical level. These ideas improved the medical development of later generation. PMID- 11877136 TI - [Probing and analyzing Zhang Jingyue's "concept of tonifying kidney's yin and yang"]. AB - Zhang Jingyue, a doctor of the Ming dynasty, is good at warming and tonifying, paying attention to tonifying the kidney. Based on the requirement of theory and practice, he held that: the method of tonifying kidney may be based on kidney -- yin; the using of bringing up yin, may be measured by containing yang, while the wonderful matter of supporting yang, may be producing yang with fostering yin. These form a unique scientific thought of "concept of tonifying kidney's yin and yang". PMID- 11877137 TI - [Studies on academic thoughts of Zhang Qi's Su wen shi yi (Interpretation of plain questions)]. AB - Summing up four academic thoughts of Zhang Qi's Su wen shi yi (Interpretation of Plain Questions), it is pointed out that his academic thought included 4 points, viz. Yang is more important than yin; the relations of yin and yang, such as interdependence, opposition, restriction were elaborated; the hepatic stagnation and splenic deficiency was the main pathogenesis of diseases; and the theory of xianghuo (ministerial fire) was developed. PMID- 11877138 TI - [Xu Shou and the provenance of "confluence of Chinese and western medicine"]. AB - Formerly, it is generally recognized that the idea of "confluence of Chinese and western medicine" was originated from Li Hongzhang's preface to the book, Wan guo yao fang (International Prescriptions) in 1890. Based on my own materials, in 1876, Xu Shou's Yi xue lun (On Medicine) already mentioned this term which may be reckoned as its earliest record. PMID- 11877139 TI - [Introduction to health education institutions in China in the first half of 20th century]. AB - The "School Hygienic Committee" established in 1929 by coordinated work of ministry of Health and Education was the earliest health education institution, promoting school hygiene education. In the same year, the "Project of School hygiene" and "Project of Health Education" were issued which were the important regulations for hygienic education in which the requirements for hygienic education unit pushed the setting up of provincial or county hygienic education committee. Based on materials from over 20 provinces and cities, about a half of the units were named health education committee, the others were called hygienic education committee, mostly attached to the Bureau of Education with its tasks aiming at students. In a few provinces and cities, health education were listed in their working program. PMID- 11877140 TI - [Eccentric therapeutic penetrating corneal transplantation]. AB - OBJECTIVE: To study the effective ways to restore sight and control the postoperative corneal astigmatism in eccentric penetrating keratoplasty. METHODS: Marks were made on the meridian where the center of recipient bed was located, and at that meridian the distance between the external edge of recipient bed and the limbus (two point marking technique) was also marked. Afterward, the donor graft was cut down from a donor eye at the corresponding position and sutured on the recipient bed. This technique was performed on 23 cases with eccentric corneal ulcer and perforation, and they were followed up for 3 months to 2 years. RESULTS: The grafts of 18 eyes remained transparent. The visual acuities were obtained > 0.1 in 16 cases, > 0.4 in 6 cases. Among them, visual acuity of 1.0 was obtained in 2 cases. CONCLUSIONS: Two point marking technique can reduce the postoperative corneal astigmatism and obtain satisfactory visual acuity in eccentric penetrating keratoplasty. This technique is simple and easy to perform. PMID- 11877141 TI - [A long-term evaluation for opticfull-thickness lamellar keratoplasty]. AB - OBJECTIVE: To investigate the long-term optic effects of optic full thickness lamellar keratoplasty (OFLK) and penetrating keratoplasty (PKP) and compare their effects. METHODS: 44 cases (48 eyes) with stromal disorder with normal endothelial function were treated by OFLK (25 eyes) or PKP (23 eyes). The optic effect of OFLK group was compared with that of the PKP group, and the endothelial density in partial cases was observed. RESULTS: The rate of transparent graft of OFLK group was 84.0%, and that of PKP group was 69.6% at postoperative 2 - 3 years, the rate of escaping blindness being 80% and 78.3% respectively. The rate of obtaining more than 0.3 vision in OFLK group was 40.0%, and in PKP group was 21.7%. Postoperatively, the endothelial cell density tended to decrease with time in PKP group, but was relatively stable in OFLK group. CONCLUSION: The results show that the optic effect of OFLK group is similar to that of PKP group, and the safety, less complications of OFLK group are the same as that of lamellar keratoplasty. It is considered that OFLK might have a bright future and the operative technique should be further studied. PMID- 11877142 TI - [Effect of keratinocyte growth factor on corneal epithelial wound healing]. AB - OBJECTIVE: To select effective methods for accelerating corneal epithelial wound healing and treating persistent corneal epithelial defect. METHODS: Exogenous keratinocyte growth factor (KGF, 0.1 ng/ml - 100 ng/ml) was added to cultures of human corneal epithelial cells and DNA synthesis in corneal epithelial cells was measured with [(3)H]-thymidine incorporation and liquid scintillation techniques. The effective concentration of topical application of KGF was calculated and 1 microg/ml KGF was then applied topically to rabbit eyes. Corneal epithelial defect was measured by a digital computer at different times. RESULTS: KGF at a dosage ranging from 1 ng/ml to 100 ng/ml enhanced cellular proliferation effectively (the rate of cell proliferation 27.66% - 76.73%) and in a dose dependent manner (r = 0.9233, P < 0.001). Topical application of KGF significantly accelerated corneal wound healing in rabbits (Epithelial healing rate in the KGF-treated group was 1.77 +/- 0.23 mm(2)/h, while in the PBS-treated group it was 1.49 +/- 0.24 mm(2)/h; P < 0.05). CONCLUSION: The results suggest that exogenous KGF stimulate proliferation of human corneal epithelial cells and accelerate corneal epithelial wound healing in vivo. PMID- 11877143 TI - [A preliminary observation on nuclear changes of human corneal endothelial cells in different kinds of corneal diseases]. AB - OBJECTIVE: To investigate the influence of different kinds of corneal diseases on corneal endothelial cells. METHODS: The endothelial cells of 91 corneal buttons obtained at the time of penetrating keratoplasty (PKP) were stained by 1% alizarin red and 0.25% trypan blue and observed under biomicroscope. RESULTS: Binuclear and polynuclear endothelial cells were found in every buttons. CONCLUSION: Under special conditions, the human corneal endothelial cells possibly have stronger ability of mitosis and compensation. PMID- 11877144 TI - [Immunopathologic studies on the effects of immunosuppressant FK 506 on penetrating keratoplasty rejection model in rat]. AB - OBJECTIVES: To study the immunopathologic process of rejection, the effect of certain immune cells and molecules on rejection and elucidate the immunosuppressive mechanism of FK 506. METHODS: By using immunohistochemical techniques, the expression of CD(4)(+) cells, CD(8)(+) cells, macrophages, IL-2 receptor, MHC-II antigen, intercellular adhesion molecule-1 (ICAM-1) and lymphocyte function-associated antigen-1 (LFA-1) in rejected corneal grafts were studied. RESULTS: Immunohistochemical studies showed diffuse infiltration of CD(4)(+) cells, CD(8)(+) cells and macrophages, and strong expression of IL-2 receptor, MHC-II antigen, ICAM-1 and LFA-1 in rejected corneal grafts, especially in the acute rejection stage. FK 506 can inhibit the expression of these cells and molecules in the grafts. CONCLUSION: Keratoplasty rejection is significantly related to delayed type of hypersensitivity and cytotoxic T cells, and immune adhesion molecules and MHC-II antigen also affect the rejection process. PMID- 11877145 TI - [Azone-induced changes on corneal endothelium in rabbits]. AB - OBJECTIVE: To investigate ocular toxic effect of azone at low concentrations in rabbits to determine whether this penetration enhancer has any value for ocular use. METHODS: 0.3%, 0.6%, 0.9% concentrations of azone in an ocular emulsion vehicle were studied. Rabbits in three experimental groups received topical 30 microl of 0.3%, 0.6%, 0.9% azone respectively three times daily for 4 weeks. Rabbits in control group 1 received 30 microl of vehicle without azone as negative control and rabbits in control group 2 received nothing as normal control. Clinical and histopathological examinations of rabbit cornea endothelial cells were made by slit-lamp biomicroscopy, light microscopy and scanning electron microscopy. RESULTS: No clinical and histopathological evidence of ocular toxicity occurred in all treated and control eyes. The concentration of azone used in this study was 3-9 times higher than that (at 0.1%) at which corneal penetration of hydrophilic compounds can be enhanced enormously. CONCLUSION: Azone has value for incorporation into ocular formulations to increase therapeutic action of many hydrophilic drugs. PMID- 11877146 TI - [Experimental study on regenerative capacity and form of corneal endothelial cells in the primate]. AB - OBJECTIVE: To investigate the regenerative capacity and form of the corneal endothelial cells of the primate. METHODS: Chromosome, HE stainings and autoradiography were made for the cornea of human and monkey. RESULTS: After trauma, corneal endothelial cells of the adult monkey in vivo showed regenerative power. The regenerative form of corneal endothelial cells of the adult, infant and fetal primate was amitosis under both physiologic and pathologic conditions. CONCLUSION: The corneal endothelial cells of the primate have regenerative power, and the form is amitosis. PMID- 11877147 TI - [Peripheral corneal edema after cataract extraction]. AB - OBJECTIVE: To report a rare type of corneal edema, i.e. peripheral corneal edema after cataract extraction---Brown-Mclean syndrome. METHODS: Clinical charts of 8 affected eyes in 5 patients were reviewed. Ultrasound pachymetry was used to determine central corneal thickness. Cell densities were measured by specular microscopy. The clinical characteristics and pathogenesis were discussed. RESULTS: The average time from operation to initial diagnosis was 10.5 years. The typical edema started inferiorly and progressed circumferentially, but central portion of the cornea was clear. The corneal thickness at edematous area was increased. The cell counts of endothelium was decreased in our patients. Additionally, the edema was associated with a punctate orange-brown pigmentation on the endothelium. CONCLUSIONS: So far this new syndrome of corneal edema has not been reported in china. Its clinical characteristics are not similar to that of conventional edema after intracapsular cataract extraction. The pathogenesis of this curious condition is still unknown. It may be associated with an aphakic eye, chronic uveitis and hereditary factors. PMID- 11877148 TI - [Determination and significance of catecholamines in aqueous humor, plasma and 24 hour urine of patients with acute angle-closure glaucoma]. AB - OBJECTIVE: To study the functional state of ocular and systemic sympathetic nervous system and the effects of its related neurohumoral factor, catecholamines (CA), on intraocular pressure (IOP) in patients with acute angle-closure glaucoma (AACG). METHODS: The levels of CA in the aqueous humor, plasma and the total amount of CA in 24 hour urine of cases with AACG during attack stage were determined with fluorometry, and the results were compared to that of patients with senile cataract and normal adults. RESULTS: The CA levels within aqueous humor and plasma and in 24 hour urine of the patients with AACG during attack stage were elevated obviously as compared with that of the control subjects (P < 0.001), and positively correlated with the level of IOP. CONCLUSION: During attack stage of AACG, the ocular sympathetic nervous system is highly excited and a large amount of CA is released into the aqueous humor, in the meantime the systemic sympathetic nervous system is also excited and CA is released into the blood circulation, which may play a certain role in the regulation of IOP. Possibly, CA is an important supplemental factor within the attack stage of AACG. PMID- 11877149 TI - [Analysis of blood flow in ophthalmic arteries in patients with middle and late stages of glaucoma with Doppler sonography]. AB - OBJECTIVE: To understand the actual blood flow in cases with middle and late stages of glaucoma and the role of vascular factor in the deterioration of visual function in such patients. METHODS: The blood flow in ophthalmic arteries was determined with color Doppler sonography (CDS) in 156 cases of patients with middle and late stages of glaucoma as well as in 75 cases of normal individuals as controls. RESULTS: The average peak systolic velocity (PSV) and end diastolic velocity (EDV) of ophthalmic artery in normal control are 35.98 +/- 9.13 cm/s and 11.98 +/- 3.90 cm/s; the PSV and EDV in primary open-angle glaucoma (POAG) group are 24.13 +/- 4.65 cm/s and 7.10 +/- 1.85 cm/s; and in primary close-angle glaucoma (PCAG) group are 22.89 +/- 5.43 cm/s and 7.06 +/- 1.92 cm/s, respectively. No significant difference was found between POAG group and PCAG group (P > 0.05). However, significant differences were shown between normal group and patients with glaucoma for both PSV and EDV (P < 0.01). CONCLUSION: The blood flow in ophthalmic arteries in patients with middle and late stages of glaucoma decreases significantly and the disorder in local blood circulation is evident, that is highly related to the deterioration of visual function and supports the vascular factor theory in the pathogenesis of glaucoma. PMID- 11877150 TI - [A study of motion perception in primary open-angle glaucoma]. AB - OBJECTIVE: To study the characteristics of the motion perception (MP) in primary open-angle glaucoma (POAG). METHOD: Forty-one patients with POAG (67 eyes) and 56 (112 eyes) normal subjects were examined by the motion perception (MP) software in PC compatible computer. RESULTS: The total abnormal rate of MP was 89.5% in POAG, and 81.6% in the early stage of POAG. By using the grade correlation analysis, there is positive correlation of the abnormal degree of MP with C/D, ocular tension, and corrected loss variance (CLV) of visualfield, negative correlation of the abnormal degree of MP with visual acuity and visualfield mean sensitivity (MS), and no correlation of the abnormal degree of MP with age and short-term fluctuation (SF) of visualfield in the POAG patients. CONCLUSION: The examination of MP is a newly reliable and simple test for the diagnosis of the early POAG. PMID- 11877151 TI - [The testing of local electroretinogram in macular diseases]. AB - OBJECTIVES: To record the local electroretinogram (LERG) in macular diseases, and define the using value of LERG in macular diseases. METHODS: The LERGs of five kinds of maculopathies were recorded with a spot light stimulator at 2.5, 5.0, 7.5 degrees of macular region. RESULTS: (1) The amplitudes of LERG at the three regions in Stargardt's disease, wet form age-related macular degeneration (AMD), central serous chorioretinopathy, and idiopathic macular hole were significantly lowered in comparison with the controls (P < 0.01). In the meanwhile, these diseases also showed high abnormal rates. While in dry form AMD and epiretinal membrane, the amplitudes of LERG had no significant changes (P > 0.05). (2) The abnormal rates of LERG were related to the stimulated area. In the three tested areas, the highest abnormal rates occurred at 2.5 degrees. CONCLUSIONS: There are mean differences in macular electrical activities in these maculopathies. LERG is an effective method of electrophysiology for testing macular function. PMID- 11877152 TI - [Application of diode laser in the operation of retinal diseases]. AB - OBJECTIVES: To evaluate the effects of diode laser photocoagulation applied on 43 cases of various kinds of fundus diseases with media opacity and on 40 cases during vitreo-retinal surgery. METHODS: Indirect ophthalmoscope laser photocoagulator was used for fundus diseases, and endolaser used during surgery. RESULTS: Vision was improved in 24.7% of the eyes, and was unchanged in 62.9% of the eyes. The neovascularization in proliferative diabetic retinopathy regressed in 78.3% of the eyes. All the retinal holes were closed. CONCLUSIONS: Diode laser can be used for treatment of some retinal diseases and in the operation. Good results can be obtained in the eyes with some extent of media opacity. No obvious side-effect was observed. PMID- 11877153 TI - [An analysis on therapeutic effects of excimer laser photorefractive keratectomy for high myopia]. AB - OBJECTIVE: To evaluate the therapeutic effects of 193-nm excimer laser photorefractive keratectomy for the correction of high myopia. METHODS: A Keracor 116 excimer laser and a tapered transition zone technique were used to treat high myopia. It was divided into two groups: the first group -6.00 - 9.75 D, 233 eyes; the second group -10.00 - 14.00 D, 55 eyes. The effect of the tapered transition zone on refraction stability was investigated in a 12-month follow-up study. RESULTS: At one year, of these eyes, 79.4% eyes attained visual acuity of 0.5 or better, 31.6% improved to 1.0 or better, 93.8% of the eyes improved or their best corrected visual acuity did not change, whereas 6.2% lost at least one line. Corneal haze was observed to be severe in one month, and diminished considerably at 6 and 12 months. At one year, no haze could be seen in 87.4%. 52.0% were within +/- 1.00 D and 80.9% in +/- 2.00 D. During one year, 14.9% were complicated with glaucoma because of using steroid drops. CONCLUSION: These data show that excimer PRK for treatment of high myopia is an effective and safe refractive surgery. For severe myopia, it is an effective surgery, but predictability is not satisfactory, and long term follow-up is necessary. PMID- 11877154 TI - [Changes in corneal sensitivity after excimer laser corneal refractive surgeries]. AB - OBJECTIVE: To observe and compare the changes in corneal sensitivity after photorefractive keratectomy (PRK) and laser in-situ keratomileusis (LASIK) for myopia. METHODS: PRK was performed on 387 eyes of 231 patients and LASIK was performed on 796 eyes of 482 patients with myopia. Corneal sensitivity was examined preoperatively and 1 week, 1, 3 and 6 months postoperatively. RESULTS: Corneal sensitivity was reduced at the early postoperative stage of both PRK and LASIK. The eyes after PRK showed greater reduction. Corneal sensitivity returned to the baseline levels at 6 months after PRK and 1 month after LASIK. CONCLUSION: The recovery of corneal sensitivity after PRK is slower than that following LASIK. PMID- 11877155 TI - [Corneal topographic analysis after excimer laser photorefractive keratectomy]. AB - OBJECTIVE: To analyze the changes in corneal topography after excimer laser photorefractive keratectomy (PRK). METHODS: The topographic changes of 122 eyes (62 patients) and 96 eyes (49 patients) on which PRK was performed were reviewed after 3 and 6 months of follow-up respectively. RESULTS: The central island pattern had a relationship with the loss of best corrected visual acuity of more than one line in early postoperative period. On subsequent follow-up, the central island pattern tended to become to other patterns (semicircular, keyhole and uniform) and the central corneal power decreased, allowing the visual acuity to improve gradually. The simulated K (Sim K) regression index was a useful indication of the changes in the corneal stroma. Sim K was related to the degree of refractive error. The greater the refractive error, the lower the Sim K. CONCLUSION: Corneal topography is a useful tool in the analysis of corneal changes in PRK. PMID- 11877157 TI - [Apoptosis in age-related macular degeneration]. AB - OBJECTIVE: To examine the role of apoptosis in photoreceptor cell death in age related macular degeneration. METHODS: 16 human eyes with age-related macular degeneration (ARMD) were studied by histopathology and TDT-mediated biotin-dUTP nick-end labeling (TUNEL) technique. RESULT: The characteristic DNA fragmentation of apoptosis was observed in scattered photoreceptor cells in 6 of 16 eyes. CONCLUSION: It is suggested that apoptosis be one of the important pathways of photoreceptor cell death in age-related macular degeneration. PMID- 11877158 TI - [Effects of freeze on Ca2+ and cyclic 3', 5'-adenosine monophosphate level in cultured human retinal pigment epithelial cells]. AB - OBJECTIVE: To investigate the mechanism of retinal pigment epithelial (RPE) cell migration and proliferation at molecular level by measuring the concentrations of intracellular cyclic 3', 5'-adenosine monophosphate (cAMP) and calcium ion (Ca(2+)) level of frozen cultured RPE cells. METHODS: The culture of human RPE cells was frozen for 0s, 15s, 30s, 45s, 60s respectively at -70 degrees C. The concentrations of cAMP and Ca(2+) in cultured human cells were measured by Fura 2/AM fluorescent technique and endogenous protein combination method. RESULTS: After freeze for 15 s, the contents of cAMP and Ca(2+) in RPE cells were not changed significantly compared with the control (no freeze). After freeze for 30s, 45s, 60s respectively, the level of intracellular cAMP was significantly decreased, while the concentration of cytoplasmic free Ca(2+) was markedly increased, showing that the concentration of cAMP and Ca(2+) being antagonistic to each other. With the re-vitalization of RPE cells for 60 minutes, the intracellular free Ca(2+) gradually reduced. CONCLUSIONS: The decreased concentration of cAMP and increased concentration of free Ca(2+) in RPE cells may be the effective mechanism of RPE cell migration and proliferation after cryotherapy, and it possibly plays an important role in the formation of proliferative vitreoretinopathy. PMID- 11877156 TI - [Corneal topography analysis after excimer laser photorefractive keratectomy]. AB - OBJECTIVE: To study the pattern of ablation, evaluate centration and stability following excimer laser photorefractive keratectomy (PRK) for myopia. METHOD: Corneal topography of 312 patients (366 eyes) with a month, 3 months and 6 months of follow-up after PRK was examined. RESULTS: The mean decentration from the pupillary center to ablating center was 0.266 mm. In both eyes, the mean decentration was located supernasally. At one month postoperative examination, uniform ablation was 49.5%, semicircular, keyhole, kidney and dumbbell shaped ablations were 42.9% and central island one was 6.0%. Central island pattern affected the best corrected visual acuity greatly. High myopia began regression at 1-3 months after PRK. CONCLUSION: Corneal topography is essential for evaluating surface changes after PRK. Long-time stability issues are answered with continued topographic follow-up. PMID- 11877159 TI - [The influence of pressure on human lamina cribrosa cell and its collagen synthesis]. AB - OBJECTIVE: To study the influence of pressure on human lamina cribrosa cell (HLC) and its synthesis of collagen. METHOD: HLCs were cultured in vitro and submitted to elevated hydrostatic pressure, their changes in morphology were observed by light and electron microscopy and their synthesis of collagen was measured with fluorescence spectrophotometer. RESULTS: Under a hydrostatic pressure of 6.67 kPa (1 kPa = 7.5 mmHg), the flat and polygonal HLCs became elongated. Vacuoles appeared in the cytoplasm and increased as the pressure persisted. The mitochondria were swollen, some of them turned into vacuoles, and myelin figures were found in the cytoplasm. When exposed to elevated pressure for 3 days, the amounts of synthesized collagen type I, III, IV were 43.95 +/- 6.37, 61.35 +/- 10.35 and 82.90 +/- 11.36 arbitrary unit (AU) by fluorometry respectively, and compared to controls, they were statistically significantly (P < 0.05) increased; after 5 days of pressure exposure, the amounts were 33.50 +/- 6.94, 40.85 +/- 12.30 and 80.45 +/- 14.65 AU, e.g., still increased. CONCLUSION: It is indicated that under pressure, HLCs undergo obvious morphological changes and increase their synthesis of collagen types I, III and IV. PMID- 11877160 TI - [An experimental study on effects of biomembrane on prevention of filtering bleb adhesion in trabeculectomy]. AB - OBJECTIVE: The study was designed to investigate the effects of biomembrane on experimental trabeculectomy. METHODS: A biomembrane was implanted under the scleral flap and the conjunctival flap in trabeculectomy in one eye of rabbits, while the fellow eye of the same rabbit without biomembrane implantation was the control. RESULTS: (1) The wound healing of the procedure reached its peak at 2-3 weeks after surgery, while the absorption of the biomembrane began at 6 weeks after surgery, and there was no tight adhesion between the biomembrane and the scar tissue, thus the filtering fistula was prevented from obstruction. (2) From the 2nd to the 7th week after surgery, the percentage of existing functional bleb in the experimental group was significantly higher than that in the control group (P < 0.05). (3) From the 1st to the 5th postoperative week, the decreasing of intraocular pressure in the experimental group was significantly greater than that in the control group (P < 0.05). CONCLUSION: The use of biomembrane in trabeculectomy can prevent the adhesion of filtering bleb and decrease intraocular pressure more effectively, and this study can provide experimental reference for the clinical use of biomembrane. PMID- 11877162 TI - [Hard nucleus cataract extraction by phacoemulsification with intraocular lens implantation]. AB - OBJECTIVE: To investigate the technique of phacoemulsification for hard nucleus cataract extraction and evaluate its therapeutic effects. METHODS: Three methods of phacoemulsification, the improved divide and conquer, the reserve lens cortex spin and no lens cortex spin, were performed on 62 eyes according to the type of nucleus hardness. RESULTS: With foldable or 5.5 mm PMMA intraocular lens implantation, the improved divide and conquer phacoemulsification was performed on 26 eyes, the reserve lens cortex spin phacoemulsification on 23 eyes and no cortex spin phacoemulsification on 13 eyes. At postoperative 3 days, 5 days and 30 days, the visual acuities without correction 0.5 or better were obtained respectively in 46 eyes (74.2%), 60 eyes (96.8%) and 61 eyes (98.4%). The major complications were corneal edema and posterior capsular rupture. CONCLUSION: The technique of phacoemulsification for hard nucleus cataract is worthy to be spread due to safety, easiness and minimum complication. PMID- 11877161 TI - [Phacoemulsification and foldable intraocular lens implantation through a clear corneal incision]. AB - OBJECTIVE: To evaluate the therapeutic effects of phacoemulsification with foldable intraocular lens implantation through a clear corneal incision. METHODS: The surgery was performed through a 3.2 mm clear corneal incision on 100 eyes of 90 patients with senile, complicated, congenital and traumatic cataracts. RESULTS: Postoperatively, the visual acuities with spherical correction or without correction were 0.5 or better in 93.0% of eyes at one day, and 1.0 or better in 56.0%, 65.1% and 67.5% at one week, one month and three months respectively. The mean postoperative astigmatism was 0.93 +/- 0.58 D at one week and 0.20 D more than the mean preoperative one (P < 0.001). There was no statistical difference between the mean preoperative astigmatism and postoperative astigmatism at one and three months which were respectively 0.78 +/ 0.60 D and 0.73 +/- 0.59 D. CONCLUSION: The phacoemulsification with foldable intraocular lens implantation through a clear corneal incision can reduce early postoperative astigmatism, shorten the period of visual rehabilitation and simplify operative procedures. PMID- 11877163 TI - [Primary posterior chamber intraocular lens implantation of posterior capsular ruptur with cataract extracted by phacoemulsification]. AB - OBJECTIVE: To evaluate the effect of primary posterior chamber intraocular lens (IOL) implantation of posterior capsular rupture (PCR) with cataract extracted by phacoemulsification. METHOD: Primary posterior chamber IOL implantation was performed on 29 cases (29 eyes) with PCR occurring in 184 cases (200 eyes) during cataract extraction by phacoemulsification. RESULTS: In-the-bag implantation was performed on 10 cases, and sulcus-fixed lens implantation on 19 cases. The post operative uncorrected visual acuities in 17 cases (58.62%) were 1.0 or better and 0.5 - 0.9 in 10 cases (34.48%). The corrected visual acuities in 22 cases (75.86%) were 1.0 or better and 0.5 - 0.9 in 5 cases (17.24%) after 3 months of operation. The complications included corneal edema and damage of pupillary margin in the operation, and anterior vitreous inflammatory reaction after the operation. CONCLUSION: The posterior chamber IOL can be primarily implanted in cases with PCR and functional anterior or posterior capsular support after the anterior segment vitreous was cleaned. PMID- 11877164 TI - [Rotated-chipping phacoemulsification in front of capsule]. AB - OBJECTIVE: The study was designed to investigate the clinical effects of rotated chipping phacoemulsification in front of capsule. METHODS: Phacoemulsification using rotated-chipping technique in front of capsule with intraocular lens implantation was performed with medium energy and high vacuum on 50 eyes. RESULTS: Rotated-chipping technique was successfully completed in 46 of 50 eyes (92%), unsuccessful cases were related to shallow anterior chamber and incomplete hydrodissection. CONCLUSION: These results suggest that rotated-chipping phacoemulsification be used for cataracts with various sclerotic nuclei and it is valuable to be applied extensively. PMID- 11877165 TI - [Congenital cataract extraction with intraocular lens implantation in children]. AB - OBJECTIVE: To evaluate the results of cataract extraction with intraocular lens (IOL) implantation in children. METHODS: Cataract extracapsular extraction with anterior vitrectomy and IOL implantation was performed on 104 eyes of 64 children with congenital cataract aged 1.5 - 14 years old. The visual acuity and possible complications were followed up for a mean period of 22 months. RESULTS: After operation 86.4% patients were relieved from blindness, 62.7% obtained corrected vision >or= 0.3, binocular vision and stereopsis were restored in 41.8% and 30.6% respectively. In 3 eyes secondary cataract developed and in 5 eyes slight iris capture occurred. CONCLUSIONS: Cataract extraction with anterior vitrectomy and IOL implantation is effective in pediatric cataract treatment. Most patients get remarkable improvement in visual acuity and visual function with minimal complications after surgery. The influence factors on the postoperative visual function need further study. PMID- 11877166 TI - [The experimental study of Nd: YAG laser injuring effects on intraocular lenses made by different materials]. AB - OBJECTIVE: To inquire the injuring effects of the Q-switched neodymium: yttrium aluminum garnet (Nd : YAG) laser on silicone, hydrogel (HEMA), acrylic, lathe cutting and cast molding polymethylmethacrylate (PMMA) intraocular lenses (IOLs). METHODS: The Nd : YAG of different energy was focused on the posterior surface of the IOL optic portion and shot in air and BSS separately. The percentages of IOL injuries were calculated, and the morphology of the IOL injuries was observed with a scanning electron microscope. RESULTS: Percentages of IOL injury: When the same energy of Nd : YAG laser was used, the injuring effects on the same kind of IOLs in air and in BSS were similar, while the laser energies of 0.5 mJ and 1.0 mJ were used, the injuring effects on different kinds of IOLs were significantly different, and when the energy of the laser was more than 1.5 mJ, the injuring effects were almost the same. Under different conditions, the morphology of the injury on different kinds of IOLs was different. CONCLUSIONS: (1) The sequence of resisting ability against Nd : YAG laser injury of different kinds of IOLs from strong to weak is: The lathe cutting PMMA IOL > acrylic IOL > cast molding PMMA IOL > HEMA IOL and silicone IOL. (2) The injuring effect of high energy Nd : YAG laser on rigid IOL is more severe than that on the soft IOL. (3) The size of IOL injuries is increased with the elevation of energy level of Nd : YAG laser. PMID- 11877167 TI - [Effects of IL-1 and TNF-alpha on the proliferation and cytosolic Ca2+ of lens epithelial cell in vitro]. AB - OBJECTIVE: To study the effects of interleukin 1 (IL-1) and tumor necrosis factor alpha (TNF-alpha) on the proliferation and cytosolic Ca(2+) of bovine lens epithelial cells (BLEC) in vitro. The effects of IL-1 and TNF-alpha on posterior capsular opacification (PCO) and the mechanism of effect on the proliferation were investigated. METHODS: The effects of IL-1 and TNF-alpha on the proliferation of BLEC were detected with MTT staining colorimetry, and cytosolic Ca(2+) was measured by fluorescence determination with Fura-2. RESULTS: IL-1 10(2) - 10(5) ng/ml, TNF-alpha 10(2) - 10(4) U/ml obviously could promote BLEC proliferation (P < 0.05 - 0.01), and increase cytosolic Ca(2+) (P < 0.05 - 0.01) at the same doses. CONCLUSIONS: IL-1, TNF-alpha contribute to the PCO. One of the mechanisms of IL-1 and TNF-alpha on proliferation is by cytosolic Ca(2+). PMID- 11877168 TI - [Assessment of six drug sensitivities in established cell lines of SO-Rb50 and SO Rb70]. AB - OBJECTIVE: With retinoblastoma (Rb) cell lines cultured in vitro to screen chemotherapeutic drugs that are effective on Rb. METHOD: Sensitivity experiments were performed with the 3, -4, 5 Dimethyliazol-2, 5 diphenyl tetrazolium bromide (MTT) assay for the drugs: dactinomycin (DACT), vincristine (VCR), VM-26, daunorubicin (DNR), DDP, bleomycin (BLMA5) against the tumor cell lines of SO Rb50 and SO-Rb70. RESULTS: The IC(50) values (microg/ml) in order at 48 hours of the above pharmaceuticals against SO-Rb50 were respectively 0.0004, 0.0016, 0.0389, 0.047, 0.29, 0.44, and those at 72 hours were 0.00025, 0.00081, 0.0151, 0.0192, 0.097, 0.11. And the IC(50) values (microg/ml) in order at 48 hours of the above pharmaceuticals against SO-Rb70 were respectively 0.00065, 0.00149, 0.0282, 0.043, 0.37, 0.215, and those at 72 hours were 0.00042, 0.00082, 0.0146, 0.0176, 0.035, 0.084. CONCLUSION: The tumor cell lines of SO-Rb50 and SO-Rb70 were sensitive to the above six pharmaceuticals. By their IC(50) values, the sensitivity order to the drugs, from higher to lower, was DACT, VCR, VM-26, DNR, DDP, BLMA5. PMID- 11877169 TI - [A study on diagnosis of ocular adnexal lymphoid hyperplasia by polymerase chain reaction amplified IgH CDR-III sequence]. AB - OBJECTIVE: To study the diagnosis and classification of ocular adnexal lymphoid hyperplasia and investigate its cell origin by using some molecular biological techniques. METHODS: 29 cases (31 eyes) of ocular adnexal lymphoid hyperplasia were examined for immunoglobulin heavy-chain (IgH) gene rearrangements by means of polymerase chain reaction (PCR) to amplify the complementarity determining region III (CDR-III) with formalin-fixed and paraffin-embedded tissue combining with histopathologic examination and immunophenotypic analysis. RESULTS: 17 eyes of malignant lymphoma and 5 eyes of reactive lymphoid hyperplasia demonstrated clear single band of the rearranged gene for the IgH chain between 100 bp. to 120 bp. The bilateral ocular adnexal lymphoma occurring simultaneously in 4 eyes (two individuals) exhibited identical band. 1 eye of malignant lymphoma, 8 eyes of atypical lymphoid hyperplasias and 6 eyes of reactive lymphoid hyperplasia showed no band or a smear band. CONCLUSIONS: The results show that this method can determine the clonality of the lymphoid hyperplasias. It may be as well in the examination of small monoclonal B cell populations for which immunophenotypical analysis is difficult. The method is an objective, accurate and useful approach for the diagnosis of ocular adnexal lymphoid hyperplasias. PMID- 11877170 TI - [The determination of neuron-specific enolase of serum in the diagnosis and supervision of retinoblastoma]. AB - OBJECTIVE: To investigate the value of determining quantity of neuron-specific enolase (NSE) in the diagnosis and supervision in patients with retinoblastoma (Rb). METHOD: We used rapid electrophoretic and the fluorometer method in the 35 serum specimens from 35 patients with Rb during surgery, after enucleation, respectively, at 3, 6, 12, >or= 18 months and 31 serum specimens from 31 control cases to determine the quantity of NSE and enolase isoenzymes. RESULTS: There were 207 +/- 65.9 u/L of all isoenzymes and 98.4 +/- 40.3 u/L of NSE from 35 patients with Rb. In comparison with the levels of control cases (respectively, 140.7 +/- 46.0 U//L of all isoenzymes, 41 +/- 10.9 u/L of NSE), both serum levels in patients with Rb increased obviously, the difference being significant. At 6, 12, and >or= 18 months after surgery, the determinations of all isoenzymes and NSE from 35 patients with Rb decreased step by step, the difference being significant in comparison with the values before the surgery. After surgery >or= 18 months, the levels of NSE and all enolase decreased to that of NSE and all enolase from control cases. CONCLUSION: The determination of enolase level, especially NSE of serum, is valuable for the diagnosis of patients with Rb. PMID- 11877171 TI - [Human trabecular cell culture in vitro]. AB - OBJECTIVE: To search the method of human trabecular cell (HTC) cultured in vitro, which could provide the necessary condition of experimental study on primary open angle glaucoma. METHOD: Organ and tissue culture were taken to get the different passages of human trabecular cells. In the meantime, human scleral fibrous cells were also cultured to contrast with HTC in morphology. RESULTS: (1) It took rather shorter time to get HTC by using organ culture method than simple tissue culture. (2) The 3rd to 5th passage of HTC was the most stable growth period, at that time they could be used in experiment. CONCLUSION: Stable growth shape and characteristic HTC could be obtained by HTC culture in vitro. PMID- 11877172 TI - [The effects of epinephrine and adrenergic antagonists on adenosine 3', 5' monophosphate level of bovine trabecular cells in vitro]. AB - OBJECTIVE: To determine the effects of epinephrine (EPI) and adrenergic antagonists on adenosine 3', 5'-monophosphate (cAMP) level of bovine trabecular cells (BTC) in vitro. METHOD: (3)H-cAMP was used in protein binding assay for measuring the intracellular level of cAMP. RESULTS: (1) 10(-5) mol/L EPI induced a fold increase of cAMP in cultured BTC in vitro; (2) Timilol and ICI 118, 551 blocked efficiently the effect of EPI at a lower concentration (10(-6) mol/L). (3) Bisoprolol did not efficiently block the effect of EPI unless at high concentrations (>or= 10(-5) mol/L). CONCLUSION: The effects of EPI increasing outflow facility may be associated with its increase of cAMP in trabecular cells; BTC contains beta-adrenergic receptors, and beta(2)-adrenergic receptors are dominant. PMID- 11877173 TI - [Clinical features of 31 cases with bilateral Duane's retraction syndrome]. AB - OBJECTIVES: To summarize the clinical features of 31 cases with bilateral Duane's retraction syndrome (DRS) and discuss its differential diagnosis. METHODS: We retrospectively summarized 31 cases with bilateral DRS from 1979 to 1996. Its clinical features including chief complaints, age and sex distribution, types of presentation, defects in abduction and adduction, retraction of the globe, upshots and downshots in abduction, etc. were analyzed. RESULTS: There were 14 males and 17 females with a female-to-male ratio 1 : 0.8. The chief complaints comprised 14 cases (45%) with ocular motility disorders and 10 cases (32%) with ocular deviations. The most common form of the syndrome was type 1 (29 cases, 94%), the remaining 2 cases (6%) with type III. Its clinical features consisted of retraction of the globe with narrowing of the palpebral fissure in attempted adduction, limitation in abduction with variable extent also in adduction, and upshot and/or downshot of the affected eye during adduction. CONCLUSIONS: In atypical cases, the retraction of the globe in adduction was not obvious and the diagnosis of DRS must be differentiated from the following ocular motility disorders, namely, abducens nerve palsy, Moebius syndrome, congenital oculomotor apraxia and congenital or infantile esotropia. PMID- 11877174 TI - [A study on the mechanism of apomorphine for treatment of myopia]. AB - OBJECTIVES: To investigate the functions of dopamine D(1) and D(2) receptor in form deprivation myopia (FDM) and the influence of apomorphine (Apo) on inhibition of FDM in chick with radioligand binding assay (RBA). METHODS: All chicks after hatching were divided into three groups: unilateral lid suture, lid suture and treated with Apo subconjunctival injections and normal control. After two weeks, they were refracted, then were killed, their equator dimensions of the eyeballs were measured quickly, the retinas were dissected out and pooled into three above separate groups. [(3)H]SCH 23390 and [(3)H] Spiperone were used for the saturation tests for D(1) and D(2) receptor protein respectively in the three groups. RESULTS: The numbers of both dopamine receptors in FDM increased (up regulation). With the treatment of Apo, the numbers of both dopamine receptors decreased (down-regulation). The affinities of both receptors were of no difference. CONCLUSIONS: The dopamine receptors in chick retina are under hypersensitive state in FDM, that is related to the biological level lowering of dopamine. Apo can inhibit the development of myopia, and the D(1) and D(2) receptor numbers are decreased, showing that Apo can affect dopamine receptors in various degrees. The method of this study can provide a route to screen drugs for treatment of myopia. PMID- 11877175 TI - [Effect of ischemia-reperfusion on membrane structure and function in rat retina]. AB - OBJECTIVE: To study the effect of ischemia-reperfusion on membrane structure and function in rat retina. METHODS: Thirty-two SD rats were randomly grouped into 4 groups. Group 1 served as the control group, group 2, 3 and 4 served as reperfusion 3 hours, 30 hours and 5 days respectively. The intraocular pressure (IOP) was raised to occlude retinal and choroidal blood vessels to form retinal ischemia; afterwards the IOP was lowered to normal to allow reperfusion. The animal was sacrificed after reperfusion. The colorimetric determination of phosphorus was used to investigate Na(+)-K(+)-ATPase activity of rat retina; the cellular membrane was labelled by fluorescence probe DPH to detect its fluidity which was measured by means of fluorescence spectrophotometer. RESULTS: Na(+) K(+)-ATPase and membrane influidity were damaged at early stage. The longer the ischemia, the more serious was the reperfusion injury. CONCLUSIONS: The mechanism of the decrease of Na(+)-K(+)-ATPase activity is due to the production of substances involved in peroxidation and excitatory amino acid (EAA) which are produced during reperfusion. They can damage the Na(+)-K(+)-ATPase directly. Lipid peroxidation disrupts membrane integrity and causes decrease of membrane influidity. For these reasons, membrane stability is reduced and Na(+)-K(+) ATPase protein function is changed. PMID- 11877176 TI - [An ultrasound biomicroscopic study on changes of the structure of ocular anterior segment after topical application of cycloplegia]. AB - OBJECTIVE: To observe the changes of ocular anterior segment and the relation between the intraocular pressure (IOP) and these changes after the topical application of cycloplegia using ultrasound biomicroscope (UBM), especially to observe the changes of ciliary body thickness and ciliary body-lens distance. METHODS: The quantitative measurement of UBM and the measurement of IOP had been done in 48 normal eyes before and after the topical use of 2% homatropine solution. The results were statistically analyzed. RESULTS: In the parameter reflecting the change of the anterior chamber angle after using 2% homatropine, the trabecular iris angle decreased, the angle open distance 250 decreased, the iris thickness 1 increased, the anterior chamber became deeper and the iris-lens contact distance became shorter, and all these differences were statistically significant (P < 0.05). In the parameter reflecting the changes of ciliary body, the ciliary body thickness and the ciliary process thickness decreased, the scleral ciliary body angle increased, the iris-zonule distance decreased and the ciliary body-lens distance increased, and all these differences were significant statistically (P < 0.05). The IOP increased from 2.35 +/- 0.46 kPa to 2.43 +/- 0.47 kPa after the topical use of 2% homatropine, but the differences were not statistically significant (P > 0.05). CONCLUSIONS: After the topical application of cycloplegia, the anterior chamber becomes deeper, the anterior chamber angle becomes narrower, the ciliary body becomes thinner and moves backward and the ciliary body-lens distance increases. By using UBM the structure of ocular anterior segment in its moving state can be observed and these structures can be measured quantitatively. The ultrasound biomicroscopic imaging has its advantages in the morphological study of ocular anterior segment. PMID- 11877177 TI - [Laser in situ keratomileusis for correction of myopia]. AB - OBJECTIVE: To evaluate the efficacy and safety of laser in situ keratomileusis (LASIK) for the correction of myopia. METHODS: A total of 429 eyes with preoperative refraction of -2.00 to -28.00 diopters were treated with LASIK and followed up for more than 12 months. Based on the refraction, the patients were divided into 3 groups: Group I, -2.00, to -8.00 diopters, 179 eyes; Groups II, 8.25 to -15.00 diopters, 137 eyes; Group III, -15.25 to -28.00 diopters, 113 eyes. The examinations included cycloplegic refractions, Goldman tonometry, corneal sensitivity, contrast sensitivity, specular microscopy, pachymetry and corneal topography. The parameters of LASIK were: the thickness of corneal cap was 130 or 160 microns, and the maximum diameter of ablation zone was 6.8 mm. After surgery, fluorometholone was given to the patients topically for 1 or 2 months. RESULTS: The vision and refractions tended to be normal 3 months after operation. The contrast sensitivity returned to normal 3 to 6 months after surgery. The endothelium did not show any changes. At 12 months, respectively 88.8%, 79.6% and 70.8% in the above groups were within +/- 1D, 73.7%, 65.0% and 46.0% were within +/- 0.5D; the uncorrected visual acuity was 0.5 or better in 96.6%, 81.8% and 54.9% of the patients, 84.4%, 58.4% and 15.0% had vision of 1.0 or better. CONCLUSIONS: LASIK is effective and safe for the correction of mild, moderate and high myopia. However, it is more complicated than PRK and more dependent on the surgical skills. PMID- 11877178 TI - [Magnetic resonant imaging diagnosis and differential diagnosis in choroidal melanoma]. AB - OBJECTIVE: To evaluate the diagnostic value of magnetic resonant imaging (MRI) for choroidal melanoma. METHODS: 12 cases of choroidal melanoma were diagnosed by means of B-ultra-sonography, computed tomography (CT), MRI and pathology. RESULTS: The MRI features of melanoma were: T(1)WI shows high signal, and T(2)WI shows low signal. CONCLUSION: Besides B-ultra-sonography and CT, MRI is also valuable in the diagnosis and differential diagnosis of choroidal melanoma. PMID- 11877179 TI - [The role of oxygen free radical in experimental keratitis]. AB - OBJECTIVE: To test a possible role of active oxygen species in corneal inflammation. METHODS: A keratitis model was induced by an intrastromal injection of lipopolysaccharide (LPS) in one eye of New Zealand albino rabbits. During the inflammation process, the activity of corneal superoxide dismutase (SOD), an enzymatic scavenger of the superoxide free radical, was measured by chemoluminescent test, the level of corneal malonaldehyde (MDA), the final product of lipid peroxidation, was determined by thiobarbituric acid test, and the clinical and histopathological changes in cornea were observed. All the results of inflammatory cornea were compared with that of saline injected cornea. RESULTS: It was found that: (1) Significant decrease in SOD activity (P < 0.001) and increase in MDA level (P = 0.001) were observed in inflammatory cornea when compared with the controls. In the period of corneal inflammation, the increase in MDA level occurred after the decrease in SOD activity. (2) The severity of corneal inflammation was negatively correlated with the corneal SOD activity (r = -0.954, P < 0.001) and positively with corneal MDA level (r = 0.734, P < 0.001). Also a statistically significant negative correlation existed between SOD activity and MDA level in the inflammatory cornea (r = -0.623, P < 0.01). CONCLUSION: These findings suggest that active oxygen species play a role in LPS induced keratitis and cause corneal injury by lipid peroxidation reactions on the polyunsaturated fatty acid of biological membranes. PMID- 11877180 TI - [Six year follow-up of suspects of primary angle-closure glaucoma]. AB - OBJECTIVE: To understand the natural history and the risk factors of primary angleclosure glaucoma and obtain experiences that might help screening for the disease. METHODS: Four hundred and eighty-five suspects were screened from 6 548 population aged over 40 years. The suspects were then followed up within six years. The following examinations were performed: visual acuity, axial anterior chamber depth, peripheral anterior chamber depth, intraocular pressure and cup/disc ratio of optic nerve papilla. The suspects with high risk were further examined. RESULTS: During the follow-up period, twenty patients (4.1%) in the suspects developed angle-closure glaucoma, 14 cases had natural attack (Acute onset developed in 6 cases and 8 cases were in chronic stage) and 6 cases were found at early stage. The anterior chamber depth shallowed progressively in 28% of suspects. CONCLUSION: Anterior chamber depth is an important index for screening of primary angle-closure glaucoma, and the regular follow-up is of significance for the suspects. PMID- 11877181 TI - [Apraclonidine prevents the acute increase of intraocular pressure in brown eyes with Nd:YAG laser iridectomy]. AB - OBJECTIVE: To investigate if apraclonidine can effectively prevent the acute increase of intraocular pressure (IOP) in brown eyes with Nd:YAG laser iridectomy. METHODS: 48 eyes with primary angle closure glaucoma were divided into treatment and control groups, which were matched by age and sex. One drop of apraclonidine was applied into the conjunctival sac at one hour before laser iridectomy and immediately after laser iridectomy in treatment group, whereas one drop of antibiotic as placebo was applied in control group at the same time. IOP, pupil diameter were measured at 0.5, 1, 1.5, 2, 3.5 hours after laser iridectomy. RESULTS: The maximum increase of IOP after laser iridectomy was 0.62 +/- 0.67 kPa and 1.13 +/- 0.87 kPa in treatment and control group respectively, there was a significant difference (P = 0.03). The obvious decrease of IOP occurred at 0.5, 1 and 1.5 hours after laser iridectomy in treatment group. No ocular and systemic side effects were found in treatment group, except pupil diameter was obviously larger in treatment group than that in control group. CONCLUSION: 1% apraclonidine effectively prevents the acute increase of IOP in brown eyes with Nd:YAG laser iridectomy. PMID- 11877183 TI - [Studies of mechanism of malignant glaucoma using ultrasound biomicroscope]. AB - OBJECTIVE: To study the mechanism of malignant glaucoma using ultrasound biomicroscope (UBM) combined with general A scan and to observe the conditions of cilio-lens block in living eyes at the realtime when malignant glaucoma is occurring. METHODS: 31 eyes from 27 patients with malignant glaucoma were evaluated and 54 normal eyes and 72 eyes of primary angle-closure glaucoma were studied as the controls. RESULTS: Of the 31 eyes, close attachments between the ciliary body and the lens were found in 18 eyes (58.1%) and slits between them were found in 13 eyes (41.9%) during the onset of malignant glaucoma. Ciliary body thickness increased significantly (P < 0.05). Fluid in the supraciliary space was found in 11 eyes. CONCLUSIONS: The mechanism of malignant glaucoma is associated with the abnormal relationship among anterior vitreous, ciliary process and lens periphery. Fluid in the supraciliary space makes the ciliary process closer to the lens periphery. It is one of the factors causing cilio-lens block. The ultrasound biomicroscopic image is a new better practicable method to diagnose malignant glaucoma during its onset compared with other methods at present used in the clinical work. It is much more valuable to differentiate the pupillary block glaucoma from malignant glaucoma by using ultrasound biomicroscopy. PMID- 11877182 TI - [A clinical observation of the therapeutic effects of pilocarpine gel for treatment of glaucoma]. AB - OBJECTIVE: To evaluate whether pilocarpine gel (PG) is better than pilocarpine solution (PS) for treatment of glaucoma. METHODS: 20 cases with glaucoma treated with 4% PG once every night were compared with 20 cases with glaucoma using 1% PS 4 times a day. The intraocular pressure (IOP), pupil diameter and refraction were examined 4 times a day in both groups. RESULTS: The PG could lower IOP by 25% - 37% and the PS by 20% - 25%, the hypotensive effect of PG could last 24 hours. The PG had the strongest hypotensive effect 5 - 13 hours after its application, and a stronger action than PS at 9 a.m. and 11 a.m, this is particularly beneficial for the control of the peak IOP before awaking and in the morning. The strongest action of miosis and accommodation enhancement of PG took place 5 - 8 hours after its application when the patients were sleeping, whereas the PS induced a fluctuating miosis and myopia in the day time. In addition, the PG was more convenient to be used than PS due to its single application for 24 hours. CONCLUSION: The PG has some advantages in the respects of compliance, lowering IOP and side-effects, thus it is worthwhile to promote its manufacture. PMID- 11877184 TI - [Ultrasound biomicroscopic dark room provocative test]. AB - OBJECTIVE: To compare the clinical application of ultrasound biomicroscopic (UBM) dark room provocative test and traditional one in screening primary angle-closure glaucoma (PACG). METHOD: 22 eyes (22 cases) with PACG in prodromal stage and 30 eyes (15 cases) with deep anterior chamber and wide angle of 15 normal persons were observed in this trial. Traditional dark room provocative test and UBM dark room provocative test were performed on all the 52 eyes at the same day time on separate days. With different positive diagnostic criteria, the sensitivity of the two methods was compared by using chi-square analysis. RESULTS: After staying in dark room for 2 hours, in PACG group, the intraocular pressure (IOP) in 10 of 22 eyes raised more than 1.07 kPa (1 kPa = 7.5 mmHg), and in 12 eyes less than 1.07 kPa or had no changes; appositional angle closure was found by Goldmann gonioscopy in 8 eyes; 3 eyes in two quadrants, 5 eyes in more than two quadrants; the appositional angle closure was found by ultrasound biomicroscopy in 15 eyes; 3 eyes in one quadrant, 5 eyes in two quadrants, 7 eyes in more than two quadrants. In the control group, IOP raised more than 1.07 kPa in 3 eyes. There was no appositional angle closure found in the control group. With the appositional angle closure as a positive diagnostic criterion, the sensitivity of the traditional test and UBM dark room test was 31.8% and 68.2%, respectively. With statistic chi-square test, there is a statistically significant difference between the two methods (P < 0.05). CONCLUSIONS: The sensitivity of UBM dark room provocative test is higher than that of the traditional one. The specificity of both methods is 100%. UBM dark room provocative test elevates the sensitivity and specificity of traditional dark room test, and reduces the false negative or positive rate in screening PACG, that is helpful to its accurate diagnosis. PMID- 11877185 TI - [An experimental study on mechanism of intraocular pressure decline after allogeneic transplantation of trabecular meshwork]. AB - OBJECTIVE: To study the mechanism of intraocular pressure (IOP) decline after allogeneic transplantation of trabecular meshwork (TM) in primate experimental glaucoma. METHODS: The experimental open-angle glaucoma models were created in 8 rhesus monkey eyes by repeated pangoniophotocoagulation on functional TM using argon laser. After 2, 3, 5 months respectively, avidin-gold or self-erythrocytes, which acted as tracers of the aqueous humor outflow, were perfused into the anterior chamber under normal pressure and volume conditions. 30 minutes later, 2.5% glutaraldehyde was perfused into the anterior chamber under the same conditions. Allograft TMs and photocoagulated area (serving as the control) were observed by light, transmission, scanning electron microscopy and ultrahistochemistry study. RESULTS: The IOP of 5 eyes of 6 experimental eyes became normal. A blood reflux into Schlemm's canal was seen by gonioscopy in 4 eyes. The surface of TMs from graft showed the normal uveal TM cords and intertrabecular spaces. Erythrocytes were found in the intertrabecular spaces and Schlemm's canal. Avidin-gold and vacuolar containing gold were observed in the cells of TM and Schlemm's canal. The TMs of photocoagulated area appeared to be fewer and narrower intertrabecular spaces. The uveal cords of this zone were confused. The content of acid mucopolysaccharides in the grafts was closer to normal than that in the photocoagulated areas. CONCLUSION: TM of allogeneic transplantation not only can survive but also function as an aqueous humor outflow pathway. We consider that this operative procedure can be used for treatment of some cases with glaucoma cautiously. PMID- 11877186 TI - [Cytotoxic effect of mitomycin C on the nonpigmented epithelium of ciliary body in rabbit eyes]. AB - OBJECTIVE: To observe pathomorphological changes of nonpigmented epithelial cells (NECs) of ciliary body after local single application of mitomycin C (MMC) during sclerectomy in rabbit eyes, and to explore cytotoxic effect of MMC on ciliary body. METHODS: A single five minutes intraoperative exposure to the agent MMC (0.2 mg/ml, 0.1 ml) was used during sclerectomy in rabbit eyes. The treated area was rinsed with 20 ml normal saline. The NECs of ciliary body at the operated area were observed under light microscope and transmission electron microscope on the 7th and 28th day postoperatively. RESULTS: On the 7th day postoperatively, the NECs were swollen, and the intracellular abundant mitochondria were swollen and vacuolized. String pearls-like membrane-encapsulated vacuoles were present in the intervals of beta-cell membrane of the NECs. On the 28th day postoperatively, these damages were gradually recovered. CONCLUSIONS: These findings suggest a toxic effect of MMC on the NECs of ciliary body, possibly that can decrease the aqueous production and intraocular pressure, even causing persistent hypotony. PMID- 11877187 TI - [Corneal epithelium implantation under corneal flap after excimer laser in situ keratomileusis: a clinical pathological and cytological study and management]. AB - OBJECTIVE: To study the clinical pathological and cytological characteristics of corneal epithelial cells implanted under the cap of laser in situ keratomileusis and the method of management. METHODS: For 4 eyes with epithelial cell implantation, the corneal cap was torn from temporal side to nasal, and the implanted material was removed from the flap and the stromal base. After washing between the interface, the cap was repositioned. RESULT: No epithelial implantation occurred again in these 4 cases after follow-up for 1 similar 12 months. The clinical pathological and cytological studies showed the material removed was degenerated and necrotic epithelial cells mixed with some unknown materials. CONCLUSION: Clinically, the material formed under the cap white and yellow in color should be removed as soon as it happens. The material consists mainly of non-structural material with necrotic and degenerated epithelial cells. PMID- 11877188 TI - [Hemorrheologic analyses of anterior ischemic optic neuropathy]. AB - OBJECTIVE: To find the pathogenic factor of non-temporal arteritis anterior ischemic optic neuropathy (AION). METHODS: Several items of hemorheology in 39 cases (19 males and 20 females) of acute non-temporal arteritis AION at papilloedema stage were examined and compared with that of the control group including normal comparable 20 males and 20 females. RESULTS: The results showed that hematocrit (HCT), fibrinogen, low shearing viscosity, aggregation index, erythrocyte electrophoresis time were higher in the AION group than that of the control group (P < 0.01 or P < 0.05). Besides, high shearing viscosity and plasma viscosity in the female patient group are also higher than that of the control group (P < 0.01). CONCLUSION: The high blood viscosity is closely related to the pathogenesis of non-temporal arteritis AION. PMID- 11877189 TI - [Evaluation of the effect of aging on the retinal nerve fiber layer thickness using scanning laser polarimeter]. AB - OBJECTIVE: To study age-related effects on the retinal nerve fiber layer thickness (RNFLT). METHODS: 75 eyes of 75 normal volunteers were recruited. Their ages ranged from 20 to 66 years. They were divided into five age groups of 15 subjects each. We measured each subject's RNFLT using scanning laser polarimeter. The mean RNFLT within the 1.5 disc diameter ring around the optic nerve papilla and its four quadrants (superior, temporal, inferior and nasal) was calculated. RESULTS: The mean +/- SD of RNFLT was 76.4 +/- 7.50 microm, 71.5 +/- 7.3 microm, 72.1 +/- 3.9 microm, 66.7 +/- 7.2 microm and 67.4 +/- 8.5 microm, respectively. CONCLUSIONS: The RNFLT decreases with aging in the normal healthy eye. PMID- 11877191 TI - [Expression of bFGF in primary human lens epithelial cells]. AB - OBJECTIVE: To study the mechanism of after cataract formation and investigate the expression of basic fibroblast growth factor (bFGF) in primary human lens epithelial cells (PHLECs). METHODS: Human lens epithelial cells were cultured in vitro, and the expression of bFGF was detected with immunohistochemistry and in situ hybridization. RESULTS: PHLECs express bFGF in protein and mRNA levels. CONCLUSION: Based on bFGF promoting PHLEC growth and differentiation, the results suggest that bFGF from PHLECs take part in promoting such cell growth and differentiation. PMID- 11877190 TI - [Relationship between changes of phospholipid and lipid peroxide of erythrocyte membrane and diabetic retinopathy]. AB - OBJECTIVE: To study the relation between the changes of phospholipid and lipid peroxide (LPD) in erythrocyte membrane in diabetes mellitus with the occurrence of diabetic retinopathy (DR). METHODS: The levels of triglyceride (TG), cholesterol (TC), glycosylated hemoglobin (HbA(1)c), phosphatidylethanolamine (PE), phosphatidylinostiol (PI), phosphatidylserine (PS), phosphatidylcholine (PC), sphingomyelin (SM), lysophosphatidylcholine (LPC), lipid peroxide (LPO) and the activity of superoxide dismutase (SOD) and glutathione peroxidase (GSH-PX) were investigated in 44 diabetic patients with DR, 20 diabetic patients with no DR (NDR) and 30 sex and age matched normal control subjects. RESULTS: A significant decrease in PE, PI, PS, PC, SM, SOD and GSH-PX and a significant increase in LPC, LPO, TG, TC and HbA(1)c were detected in DR and NDR group when compared with the normal controls. These changes were more obvious in DR group. CONCLUSION: The lipid peroxidation and the alteration of the erythrocyte membrane phospholipid compositions may contribute to the occurrence and development of DR. PMID- 11877192 TI - [A study on the prevention of selenite cataract with taurine]. AB - OBJECTIVE: To evaluate the prevention of selenite cataract with taurine. METHODS: A prior injection of taurine was given intraperitoneally to Wistar rats 2 days before the selenite challenge and a following daily injection of taurine was needed until the experiment was finished. All of the rats were examined under a split-lamp microscope on selected days. RESULTS: 0.5% taurine showed no effect on lenses, 1%, 2% and 4% taurine could delay the development of nuclear opacity, and 2% and 4% taurine could also delay the development of cortical opacity. CONCLUSIONS: Taurine can prevent the development of selenite cataract. Its effect is dose-dependent. PMID- 11877193 TI - [Intraocular penetration of topical lomefloxacin in rabbit's eyes]. AB - OBJECTIVE: To measure the rabbit ocular tissue concentration of lomefloxacin (LMLX) after its topical application. METHODS: Fifteen rabbits (30 eyes) for the experiment (12 white rabbits and 3 pigmented rabbits) were divided into 5 groups. In the 1st group, 0.3% LMLX eyedrops were instilled once (1 drop each) in the 3 white rabbit's eyes. In the 2nd group, 0.3% LMLX eyedrops were instilled for 6 times (1 drop once) with an interval of 5 minutes in the 3 white rabbit's eyes. In the 3rd group, 0.3% LMLX eyedrops were instilled once (1drop each) in the 3 white rabbit's eyes after removal of corneal epithelium. In the 4th group, 0.3% LMLX eyedrops were instilled once (1 drop each) in the 3 pigmented rabbit's eyes. In the 5th group, LMLX 0.5 mg was subconjunctivally injected in the 3 white rabbit's eyes. After the administration for 30 minutes, aqueous humor was collected and the eyeball was enucleated. All measurements were performed by high pressure liquid chromatography (HPLC). RESULTS: In the 1st group, the mean levels of drug were 8.75 +/- 3.09 microg/g in cornea, 0.21 +/- 0.1 microg/ml in aqueous humor, and 1.49 +/- 0.39 microg/g in iris, respectively. The ocular tissue concentrations of the other groups were significantly higher than that of the 1st group. The pigmented tissue of pigmented rabbit's eyes had relatively high concentration of LMLX. CONCLUSION: All concentrations of LMLX in rabbit's eyes are above the minimum inhibitory concentration for 90% (MIC(90)) of most bacterial pathogens. LMLX has greater affinity in the pigmented tissue. These results show that LMLX may be an effective antimicrobial agent for prophylactic use in ophthalmic surgery and also for the treatment of intraocular infections due to susceptible organisms. PMID- 11877194 TI - [An experimental research of recombinant human epidermal growth factor on corneal wound healing]. AB - OBJECTIVE: To investigate the effects of recombinant human epidermal growth factor (rhEGF) eye drops on corneal wound healing. METHODS: Twenty-four white rabbits were randomly divided into 4 groups, 6 rabbits 12 eyes each. Anterior keratectomy of 8 mm in diameter and 1/3 cornea in thickness was performed on each eye. Each of the following concentrations of rhEGF: 1, 10, 100 microg/ml eye drops or normal saline (control) was applied four times daily for a week respectively for one group. The wound area was determined by computer imaging analysis. RESULTS: The mean epithelial healing rate of rhEGF 1, 10, 100 microg/ml groups was 9.31, 9.96, 9.31 mm(2)/day respectively, significantly greater than 8.11 mm(2)/day of the control group. The action of rhEGF of 10 microg/ml was somewhat better than that of 1 or 100 microg/ml, and no significant difference was noticed among the three rhEGF groups. Moderate inflammation and corneal neovascularization were induced in the rhEGF 100 microg/ml treated group. CONCLUSION: rhEGF 1 - 10 microg/ml can accelerate corneal wound healing in the rabbit with no adverse side-effects. It may be used to treat serious corneal trauma and ulcer clinically. PMID- 11877195 TI - [An experimental study on formation of cellular membrane on the surface of implanted posterior chamber intraocular lens]. AB - OBJECTIVE: To observe the cellular reaction and membrane formation on the surface of intraocular lens (IOL) implanted in rabbit eyes and discuss the mechanism of early postoperative membrane formation and biological action. METHODS: 20 adult pigmented rabbit eyes were given posterior chamber IOL implantation. IOLs were extracted on the postoperative 1st, 3rd, 7th, 14th and 28th days respectively. Samples were observed under light microscope and scanning electron microscope (SEM). RESULTS: Macrophages on the surface of IOL had transformed into fibroblast like cells and the formation of eosinophilic membrane had occurred on the 3rd day after operation with tiny thread-like things interwoven among cells forming net like supporting membrane under SEM. CONCLUSION: It infers that the membrane is a kind of fibrinous film secreted by fibroblast-like cells and the cells involved in reaction are the biological foundation of membrane formation; this membrane will prohibit the serious cellular reaction at the end and keep IOL transparent. PMID- 11877196 TI - [A study of lipid peroxide-induced damage and of ultrastructure of lens epithelial cells in lens organ culture in vitro]. AB - OBJECTIVE: To understand the mechanism of the rabbit lens damage from lipid peroxidation (LPO) involved in cataract development. METHODS: Ultraviolet (UV) radiation was used to induce the damage of peroxidation on cultured rabbit lens. Cultured rabbit lenses were exposed to UV radiation in MEM cultural medium without or with superoxide dismutase (SOD) added at 10 U/ml. The thiobarbituric acid (TBA) colorimetric method was used to detect the lens malondialdehyde (MDA) content produced by LPO, and electron microscopy was used to observe the ultrastructure of rabbit lens epithelium. RESULTS: In the lens with UV radiation, MDA was increased as compared with that in non-irradiated one (P < 0.01) and in UV + SOD group (P < 0.01). The membranes of lens epithelial cells were damaged and mitochondria, etc. membranous structures disappeared with clumping pattern of chromatin, while the cells in UV + SOD and control groups appeared normal in structure. CONCLUSION: UV radiation can induce lipid peroxide damage in lens culture of rabbit. PMID- 11877197 TI - [The survey of anterior chamber angle in a population of middle and old age]. AB - OBJECTIVE: To determine the condition of anterior chamber angle in a population of middle and old age, and afford data for screening of angle closure glaucoma. METHODS: Two hundred people aged 50 years or older were selected by random sampling in four districts of Doumen county. The anterior chamber angle of all people was evaluated by Goldmann one-mirror gonioscope, and the angle was graded according to the Spaeth and Shaffer's grading system. RESULTS: Three hundred ninety eyes were examined, 12% (inferior angle) to 30% (superior angle) were Shaffer's grade 0.05). CONCLUSION: 8-Cl-A can inhibit the three growth factor-induced proliferation of RPE cells. PMID- 11877207 TI - [An experimental study of human trabecular cell growth on a filter and hydraulic conductivity]. AB - OBJECTIVE: To search a method of culturing human trabecular meshwork cells (HTC) on a filter support and provide a model to study the hydraulic conductivity of HTC in vitro. METHODS: The 3rd passage of HTC was cultured on a nylon filter, after that we measured the rate of different irrigation through the filter with HTC [Lp, microl/(min x mmHg x cm(2))]. RESULTS: HTC could continuously grow on the filter. The normal Lp was 10.45 microl/(min x mmHg x cm(2)). When the model was irrigated respectively by the solution containing epinephrine (EPI) and dexamethasone (DEX), the Lp value of HTC cultured in the same period was significantly higher than that of the normal group. However, after HTC exposed to DEX for a few days, Lp was significantly decreased. CONCLUSIONS: (1) Better information of hydraulic conductivity and the effects of medicine on HTC can be obtained from the dynamic filter model; (2) EPI can improve the conductivity of HTC, and DEX can do the same in early period. PMID- 11877206 TI - [Immunohistochemical studies on wholemounts of the cornea and iris-ciliary body after corneal transplantation]. AB - OBJECTIVE: To study the phenotype of the cells involved in the pathogenesis of allograft rejection after corneal transplantation. METHODS: Wholemounts of the cornea and iris-ciliary body were isolated and prepared from normal rats and those after penetrating corneal transplantation. Immunohistochemical stainings were carried out on these wholemounts using monoclonal antibodies to CD3 (T cell), CD4 [T-helper (Th) cell], CD8 [T-suppressor (Ts) cell], macrophages, dendritic cells, B lymphocytes, major histocompatibility complex (MHC) class II antigen and transforming growth factor-beta (TGF-beta). RESULTS: All the cells mentioned above except B lymphocytes were positive and present in a small number in the peripheral cornea and limbus of normal rats. A massive influx of CD3, CD4, CD8 positive cells, macrophages, dendritic cells, MHC class II and TGF-beta positive cells not only into cornea but also into the iris was observed 7 days after allograft corneal transplantation. Whereas no B cell was found in the allograft. Such a cellular response was not found in the rats in which corneal transplantation with autologous cornea was performed. CONCLUSION: T cells, Th cells, Ts cells, macrophages, dendritic cells, MHC class II positive cells and TGF-beta positive cells are involved in the pathogenesis of corneal allograft transplantation. The involvement of the iris and ciliary body may promote or aggravate the allograft rejection. PMID- 11877209 TI - [The manifestations of 496 cases with thyroid-related immune orbitopathy and normal thyroid function]. AB - OBJECTIVE: To observe the clinical manifestations of thyroid-related immune orbitopathy (TRIO) with normal thyroid function. METHODS: 496 cases with TRIO were collected from 1973 to 1994 and their clinical manifestations were analyzed. RESULTS: The clinical manifestations showed that males are more than females. The disease occurred in middle aged persons with unilateral exophthalmos as the primary early manifestation. Most of the cases had the signs of eyelids and extraocular muscle involvement. CONCLUSIONS: The thyroid-related immune orbitopathy with normal thyroid function can be named thyroid-related immune orbitopathy-II. The clinical characteristics of this disease are unilateral, of signs of eyelids, exophthalmos and enlargement of eye muscles. The pseudo-tumors of the orbit are mainly considered for differential diagnosis. PMID- 11877208 TI - [Culture of human trabecular meshwork cells and the cell characteristics of immunohistochemical studies]. AB - OBJECTIVE: To look for better cultural methods in order to obtain numerous human trabecular cells in vitro for glaucoma experimental studies, and describe the immunohistochemical characteristics of the cells. METHOD: Human trabecular meshwork cells were cultured, then 4 monoclonal antibodies were used for immunohistochemical stains of the cultured cells. RESULTS: At the primary period, the growth of human trabecular cells was obviously slower than that of cows and pigs. The immunohistochemical stains showed that the cells presented positive reactions to neuronal specific enolase (NSE) and vimentin and negative reactions to factor VIII related antigen (VIIIR:Ag) and desmin. CONCLUSIONS: The culture of human trabecular meshwork cells in vitro needs more careful and better cultural conditions. The cells originally are derived from embryonic neural crest, not from mesodermal endothelium of blood vessels. There is middle filament vimentin and no desmin in the cells. PMID- 11877210 TI - [Excimer laser in situ keratomileusis for treatment of myopia]. AB - OBJECTIVE: To evaluate the effectiveness of excimer laser in situ keratomileusis (LASIK) for treatment of myopia. METHODS: 928 eyes of 678 myopic patients who received LASIK were studied. The patients were divided into 3 groups according to preoperative diopters (-2.00 - -6.00D, -6.10 - -10.00D & -10.10 - -20.00D). The mean follow-up was 12 months (6 - 18 months) and the results were statistically analyzed. RESULTS: 12 months after surgery, in the first group the mean residual refraction was -0.24 +/- 0.50D in 86% and 92% of eyes were in the ranges of refraction +/- 0.50D and +/- 1.00D, the uncorrected visual acuities of 0.5 and 1.0 were in 99% and 86% of the eyes respectively. In the second group, the mean residual refraction was -0.47 +/- 0.64D, the refraction of 83% and 90% of eyes was in the ranges of +/- 0.50D and +/- 1.00D, and the uncorrected visual acuities in 95% and 79% of eyes were 0.5 and 1.0 respectively. In the third group, the mean residual refraction was -0.89 +/- 1.02D, the refraction of 36% and 62% of the eyes were in the ranges of +/- 0.50D and +/- 1.00D, and the uncorrected visual acuities in 68% and 45% of the eyes were 0.5 and 1.0 respectively. 47 eyes had been retreated with LASIK. 5 eyes lost two or more lines of the best corrected visual acuity due to irregular astigmatism, decentration and progressive myopic maculopathy. CONCLUSION: The results indicate that LASIK can treat myopia in the range of -2.00 - -20.00 D effectively without vision threatening complications. Some modification of the surgical techniques and laser nomogram will help to improve predictability and stability of outcome. PMID- 11877211 TI - [Treatment of intraocular fibrinous membrane by anterior chamber injection of tissue plasminogen activator after IOL implantation]. AB - OBJECTIVE: To investigate the method of treatment of intraocular fibrinous membrane by tissue plasminogen activator (t-PA) after intraocular lens (IOL) implantation and evaluate its clinical results. METHODS: 58 eyes of 43 patients after cataract surgery were divided into three groups, and received t-PA anterior chamber (AC) injection or combined with YAG laser therapy at different times after the operation. Group 1 included 26 eyes which received 4 microg t-PA AC injection at the end of operation. In group 2, 22 eyes whose fibrinous membrane was formed 5 - 7 days after the operation were injected with 4 microg t-PA at that time. In group 3, 10 eyes with thicker membrane were given YAG laser treatment at postoperative 5 - 9 days and then 4 microg t-PA was injected into the AC. RESULTS: In group 1, hyphema occurred in 4 eyes, in 8 eyes at the pupillary region the fibrinous membrane was formed which was completely lysed after t-PA AC injection at postoperative 3 - 5 days. In group 2, the membrane was lysed within 24 hours in 20 of 22 eyes and no hyphema was found. In group 3, the membrane was lysed within 4 - 24 hours after t-PA injection in all cases, and no hyphema occurred. CONCLUSION: It is very effective to treat fibrinous membrane formation after IOL implantation by t-PA AC injection at postoperative 1 week when the AC inflammation is well controlled. The combined use of YAG laser with t PA AC injection is much more effective for the treatment of such a membrane. PMID- 11877212 TI - [Phacoemulsification in myopia and negative or low powered posterior chamber intraocular lens implantation]. AB - OBJECTIVE: The study was designed to investigate the technique and clinical experience of phacoemulsification in myopia and negative or low powered posterior chamber intraocular lens implantation (PC-IOL). METHODS: 120 eyes of 102 cases with myopia and cataract were included (axial length >or=28 mm) and hardness of nucleus was of grade 2 in 5 eyes, grade 3 in 84 eyes, grade 4 in 16 eyes and grade 5 in 15 eyes. Tunnel incision was made and in situ nuclear fracture technique by phacoemulsification was used. Energy and time of emulsification: 50% - 60% of energy was set up and emulsification spent 40s to 2 min. 10 s (mean, 1 min. 11.3s) for nuclear sclerosis of grade 2 or 3.65% - 70% of energy was set up and emulsification spent 48s to 1 min. 22s (mean, 68.7s) for cutting the central nucleus and removing the divided nucleus (grade 4 or 5). RESULTS: There were 120 eyes in this group. Eleven to 22 months after operation (mean 14.5 months) the visual acuities of 0.5 or better with and without correction were obtained in 62.6% and 43.4% respectively. The major complications were corneal edema and rupture of posterior capsule. No retinal detachment and cystoid macular edema were found. CONCLUSION: This operation is suitable for myopia with cataract. It is worth to spread due to good visual result, proper correction of ametropia, prevention of retinal detachment, and decrease of operative complications. PMID- 11877213 TI - [Determination of posterior lens capsule status in traumatic cataract with B ultrasonography]. AB - OBJECTIVE: To determine the status of posterior lens capsule in traumatic cataract with B-ultrasonography. METHODS: 44 eyes of traumatic cataract were studied. The integrity of posterior lens capsule was evaluated with B ultrasonography before surgery. Cataract extraction was performed under microscope and the status of posterior lens capsule was recorded. RESULTS: 21 eyes showed complete posterior lens capsule in B-ultrasonography. Of them, 19 eyes (90.5%) were confirmed by surgery. Other 23 eyes with posterior lens rupture determined before operation were all confirmed by surgery (100.0%). CONCLUSION: B ultrasonography is helpful in evaluating the integrity of posterior lens capsule and choosing the strategy of surgery. PMID- 11877214 TI - [Light sensitivity in central visual field]. AB - OBJECTIVE: To examine the difference in the light sensitivity in the central visual field between the normal subjects and glaucoma patients with or without the early visual field damages and propose the criteria to identify the possible early disturbance in the visual field. METHODS: The light sensitivity at the 76 locations in the central field was measured in 108 normal eyes of 75 normal volunteers aged from 20 to 78 years and 82 eyes of 53 patients with early open angle or angle-closure glaucoma aged from 30 to 70 years on the Humphrey Field Analyzer (Model 610) by using the Central-30-2 program. RESULTS: No sex and eye differences in the light sensitivity were found between the two groups. The significant difference in the light sensitivity existed between the age of 20 - 45 years and the age of 46 - 78 years in the normal subjects. The lower limit of 80% normal range for each location in the central field of the two age groups was calculated. The light sensitivity decreased by 2 - 3 dB at every 10 degree from the eccentricity. The light sensitivity in the inferior field was 1 - 2 dB higher than that in the superior. There was no statistical difference between the nasal and the temporal field. For the patients with the field defects the light sensitivity of many locations decreased by 4 - 8 dB, and by 1 or 2 dB in the patients without the field defects as compared with the normal control. CONCLUSIONS: Based on the comparison with the normal control and the lower limit of 80% normal range, we propose the following criteria for classification of damages in the glaucoma visual field. Grade I: light sensitivity decreases by 1 - 3 dB at a cluster of locations in the field; Grade II: light sensitivity reduces by 4 - 6 dB at a cluster of locations in the field; Grade III: light sensitivity reduces by more than 6 dB at a cluster of locations in the filed. PMID- 11877215 TI - [Effects of homoharringtonine liposomes and homoharringtonine solution on glaucoma filtration surgery in rabbits]. AB - OBJECTIVE: Homoharringtonine (HHT), a Chinese herbal drug, was used in the form of liposome or solution as an adjunct to glaucoma filtration surgery in rabbits to determine its effect in preventing closure of sclerostomy. METHODS: Forty rabbits that underwent a standard posterior-lip sclerectomy in both eyes were divided into 5 post-operative groups. Group 1 received subconjunctival HHT liposomes (0.025%). Group 2 received subconjunctival blank liposomes and served as control. Group 3 and 5 received subconjunctival HHT solution at low (0.025%) and high (0.1%) doses respectively. Group 4 received subconjunctival normal saline and served as control. Intraocular pressure, interval between operation and bleb failure, rate of scleral fistula occlusion and ocular toxicity were determined in each group. RESULTS: Subconjunctival treatment with HHT solution at a high dose transiently prolongs the survival of filtration surgery, but the subconjunctival use with HHT liposomes or HHT solution at a low dose does not significantly prolong the survival of filtration surgery in rabbits. Significant corneal haze and corneal neovascularization resulted from using HHT at a high dose. CONCLUSIONS: The success rate of rabbit filtration surgery can not be markedly increased by the postoperative use of HHT liposomes or its low dose solution. When the high dosage of HHT solution is used, transient elevation of the success rate can be obtained, however the side-effect is also significantly increased. PMID- 11877216 TI - [The choice of methods of intraocular lens implantation in traumatic cataract in the absence of capsular and zonular support]. AB - OBJECTIVE: To study the choice between the methods of anterior chamber intraocular lens (AC-IOL) implantation and posterior chamber IOL suture fixation in traumatic cataract without posterior capsular support. METHODS: The study included 86 cases (86 eyes) with traumatic cataract. A new flexible open-loop one piece PMMA IOL was implanted in the AC-IOL group of 56 cases (56 eyes); fixation of a C-loop one piece PMMA IOL at ciliary sulcus with two sutures was performed on 30 cases (30 eyes) in the posterior chamber IOL (PC-IOL) group. RESULTS: The follow-up period was 1 - 42 months. The visual acuity 0.5 or better was in 71.4% (40/56) in AC-IOL group, in 90.0% (27/30) in PC-IOL group; 1.0 or better was in 39.3% (22/56) in AC-IOL group, in 46.7% (14/30) in PC-IOL group. CONCLUSION: Both AC-IOL implantation and PC-IOL suture fixation are the remedy methods when the PC IOL intracapsular implantation is unsuccessful. The AC-IOL is suitable for old persons with good anterior segment (cornea, iris and the angle of anterior chamber) and poor posterior segment (vitreous, retina) situations, while the PC IOL suture fixation is suitable for the young with relatively poor anterior segment and good posterior segment conditions. PMID- 11877218 TI - [Effects of tripterygium wilfordii polyglyco - sidium on intraocular inflammatory reaction after intraocular lens implantation]. AB - OBJECTIVE: The effects of tripterygium wilfordii polyglyco - sidium (TWP) on intraocular inflammatory reaction after extracapsular cataract extraction and intraocular lens (IOL) implantation were studied. METHODS: 120 cases with senile cataract were divided into two groups: (1) Patients received TWP after IOL implantation; (2) patients received prednisone after IOL implantation. The slit - lamp was used to observe inflammatory cells and fibrinous exudate in the aqueous humor and cellular deposits on IOL on the 1st, 3rd, 7th and 14th postoperative days. RESULTS: The study indicated that on the 1st, 3rd, 7th and 14th postoperative days the inflammatory cells and fibrinous exudate in the aqueous humor and cellular deposits on IOL were significantly lower in TWP - treated group than that in steroid - treated group (P < 0.05). CONCLUSIONS: TWP is effective in reducing postoperative anterior ocular inflammation after extracapsular cataract extraction and IOL implantation. The TWP is more effective than the steroid in suppressing postoperative intraocular inflammatory reaction. PMID- 11877217 TI - [Observation of anterior chamber angle with ultrasonic biomicroscopy after posterior chamber implantation of intraocular lens]. AB - OBJECTIVE: To observe the changes of anterior chamber angle after extracapsular cataract extraction (ECCE) with posterior chamber implantation of intraocular lens (IOL) and discuss the associated factors. METHODS: The changes of anterior chamber angle in 31 cases (41 eyes) with senile cataract were observed with ultrasonic biomicroscopy after ECCE and posterior chamber implantation of IOL. RESULTS: (1) There were 6 eyes with peripheral anterior synechia of the iris, 12 eyes with abnormal pupillary configuration, which were related to the position of IOL haptics. (2) The IOL haptics fixed out of the capsular bag were not all fixed at the ciliary grooves. (3) After postoperative 3 months, the residual cortex of the lens still existed. Too much residual cortex in the eye caused postoperative peripheral iris bombe and narrowing of chamber angle. CONCLUSION: The changes of anterior chamber angle after posterior chamber implantation of IOL are related to the position of IOL haptics and much residual cortex must be avoided. PMID- 11877219 TI - [Phacoemulsification for mature or hypermature cataracts]. AB - OBJECTIVE: To discuss how to choose the methods of capsularhexis and phacoemulsification in cases with mature or hypermature cataract. METHODS: On fifty-nine mature or hypermature cataract patients, continuous circular capsularhexis or can-opener capsulotomy was performed. The nucleus was emulsified by the method of divide and conquer or phaco chop or rotatory carousel in the bag or at the plane of iris. RESULTS: Continuous circular capsularhexis was successful in 51 cases, failed or shifted to can-opener in 8 cases. There were posterior capsular rupture in 1 cases, partial dehiscence of the zonule in 1 cases and iris bitten by the phaco tip in 1 cases. Postoperatively, there were mild corneal edema in 6 cases and iris posterior synechia in 1 cases. CONCLUSION: Continuous circular capsularhexis is the key point to obtain a successful phacoemulsification for mature or hypermature cataract. Hydrodissection is not important before the phacoemulsification. Skillful phaco chop can reduce the surgical manipulation and the postoperative reaction. PMID- 11877220 TI - [Intraocular pressure rise early after phacoemulsification with posterior chamber lens implantation]. AB - OBJECTIVE: To investigate intraocular pressure (IOP) changes early after phacoemulsification with posterior chamber lens implantation. METHODS: The IOP was measured in 49 patients 53 eyes before and 10 - 12, 22 - 24, 46 - 48 hours after the operation. RESULTS: The IOP increased obviously in all eyes 10 - 12 hours after the operation, and rose more significantly from the baseline to the maximum 10 - 12 hours after surgery in 20 sutured eyes than that in 33 unsutured eyes (t = 4.41 and 3.32, P = 0.0001 and 0.0020, respectively). It was higher than 3 kPa in 16 eyes, 10 (10/20, 50.0%) in the sutured group, 6 (6/33, 18.2%) in the unsutured group, respectively, and the difference was significant between the two groups (chi2=5.9820, P=0.014). The IOP decreased markedly 22 - 24 hours, and returned to the baseline level 46 - 48 hours after the operation. CONCLUSION: These results indicate that IOP increases early after phacoemulsification with posterior chamber lens implantation, especially in sutured eyes. Therefore, to apply corresponding management properly in time and obtain better therapeutic effects of the operation, IOP should be measured regularly. PMID- 11877221 TI - [The choice of intraocular lens before cataract extraction for cases with high myopia]. AB - OBJECTIVE: To study the choice of diopter and size of optical portion of intraocular lens (IOL) before cataract extraction for cases with high myopia. METHODS: Extracapsular cataract extraction (ECCE) or phacoemulsification with IOL implantation was performed on 60 cases (94 eyes) with high myopia (axial length > 26mm). The posterior chamber IOL selected and inserted was the one with the diopter and size of the optical portion suitable for the recipient to have a postoperative low myopia and easy to be examined with an ophthalmoscope. RESULTS: The postoperative eyes were of low myopia, -2.07 +/- 1.95 D. The larger size of optical portion was more suitable for ocular fundus examination. Great care was taken for phacoemulsification for cataractous nucleus below III grade. CONCLUSIONS: For patients with axial myopia, the IOL implanted should create low myopia postoperatively. Larger optic size is better for fundus examination. Phacoemulsification can produce less astigmatism and damage, but the indication must be controlled strictly. PMID- 11877222 TI - [Residue of sodium hyaluronate in aqueous humor following intraocular lens implantation]. AB - OBJECTIVE: To determine the residue of sodium hyaluronate in aqueous humor following intraocular lens (IOL) implantation. METHODS: The percentage concentration of sodium hyaluronate in aqueous humor in 23 patients (25 eyes) having undergone IOL implantation was measured by Shimadzu UV-3101 ultraviolet spectrophotometer before and after irrigation with BSS 2 ml, 5 ml and 8 ml. RESULTS: The different percentage concentrations of sodium hyaluronate were 65.50% +/- 18.12%, 10.20% +/- 4.80%, 2.19% +/- 1.83% and 0.70% +/- 0.91%. CONCLUSION: Replacement of aqueous humor by BSS 2 ml can eliminate the most part of sodium hyaluronate. Replacement of aqueous humor by BSS 8 ml can decrease the residue of sodium hyaluronate to trace amount and the postoperative intraocular pressure can maintain in normal range. PMID- 11877223 TI - [Experimental observation of foreign gene transfer with lipofectin to the primary human lens epithelial cells]. AB - OBJECTIVE: To determine whether an exogenous gene carried by lipofectin can be introduced into the primary human lens epithelial cells. METHODS: Plasmid DNA with beta-galactosidase gene carried by lipofectin was applied to primary cultured human lens epithelial cells. Gene expression was detected by enzymatic color reaction using X-gal as a substrate in the 2 and 6 days of expression after 12, 24, 36 hours of transfection respectively. The gene transfer positive rate of the lens epithelial cells was counted. RESULTS: The foreign gene could be transferred into primary human lens epithelial cells by lipofectin. In the expression of 2 days, transfer positive rate was up to 48% after 24 hours of transfection. CONCLUSIONS: Efficient and stable transfer of the functional gene can be achieved by lipofectin into the lens epithelial cells. Lipofectin is an available and a promising vehicle for delivering aim gene and studying on the mechanism of physiology and pathology of lens epithelial cells. PMID- 11877224 TI - [The experimental research of free calcium of the lens epithelial cells in galactose cataract]. AB - OBJECTIVE: To study the changes of free calcium in the lens epithelial cells (LECs) of galactose cataract. METHODS: Wistar rats, 28 days old, were fed with 50% galactose forage to form cataract models. Then the concentration of free calcium of LECs was measured at the 1st, 3rd, 7th, 10th, 14th, 19th days after the galactose diet was given. RESULT: The concentrations of free calcium in LECs of galactose cataract increased apparently and reached the maximum at the 10th day after galactose diet. The maximum was 4 - 5 times higher than that of the normal group. CONCLUSION: The calcium concentration of LECs in galactose cataract increases but the degrees at different phases are not the same. The maximal concentration is at the phase when the cortex is opacified. PMID- 11877225 TI - [Cell density and expression of proliferating cell nuclear antigen (PCNA) in the lens epithelium of children and aging patients with cataract]. AB - OBJECTIVE: To determine the cell density and expression of proliferating cell nuclear antigen (PCNA) in the lens epithelium of children and aging patients with cataract. METHODS: The lens epithelial cells (LECs) of the central area were examined in two groups, children (less than 12 years old) and senile persons (51 80 years old). The densities of LECs in the two groups were calculated with samples stained with hematoxylin and eosin (HE) staining, and the PCNA was tested immunohistochemically. The integral optical density (IOD) of PCNA expression was measured through a medical color image analysis system. RESULTS: The densities of LECs in children and senile persons were 5,020.25 +/- 246.01 cells/mm(2) and 4,340.00 +/- 240.95 cells/mm(2) (P < 0.05), the IODs were 2.89 +/- 0.57 and 2.13 +/- 0.63 (P < 0.05) respectively. CONCLUSION: The LEC counting and IOD values in children eyes are significantly higher than that of senile persons, suggesting that they be one of the important factors in the formation of posterior capsular opacification (PCO) or after cataract in children eyes. PMID- 11877226 TI - [A study on lens epithelial cell ultrastructure in rhesus monkey cataract]. AB - OBJECTIVE: To investigate the ultrastructure of lens epithelial cells of normal eyes and cataractous eyes in rhesus monkeys. METHODS: Anterior capsules were taken from 4 normal lenses and 4 cataractous lenses of rhesus monkey eyes, and the samples were observed under transmission electron microscope. RESULTS: The ultrastructure of the cataractous lens epithelial cells changed obviously. Mitochondria with disappearance of crista were swollen, and vacuolar. Hollow cell with a single large vacuole in the cytoplasm could be seen. The cytoplasm presented dissolution. Cells were swollen and damaged, even cell membrane disruption could be seen. Karyopyknosis, nuclear deformity, disappearance of perinuclear cistern and nuclear pores could be found. The heterochromatin was concentrated and aggregated at the nuclear periphery. CONCLUSIONS: The ultrastructural changes of the lens epithelial cells in monkey cataractous lens are similar to that in human cataractous lens. Rhesus monkey cataract may be a suitable experimental model to investigate the formation of human cataract. PMID- 11877227 TI - [A randomized control clinical trial of glaucoma filtering surgery with homoharringtonine]. AB - OBJECTIVE: To study the antiproliferative effects of homoharringtonine (Hh) on glaucoma filtering surgery. METHODS: In a randomized control clinical trial, 78 patients (88 eyes) with refractory glaucoma underwent trabeculectomy with and without Hh. In the Hh-treated eyes (n = 46), the therapeutic dose of Hh was: intraoperative application of Hh 0.4 mg and postoperative subconjunctival injections of Hh 0.62 +/- 0.20 mg (ranged 0.53 - 0.75 mg). In the control eyes (n = 42), Hh was not used. The follow-up period was 18 to 48 months, and the data were analyzed by using the life-table method of Kaplan-Meier. RESULTS: The cumulative success probability in Hh group was 84.5% and that in control group was 50.9%, the difference being significant (P < 0.05). The cumulative percentage of functioning bleb in Hh group was 84.2% and that in control group was 52.9% (P < 0.05). The rates of corneal erosion were 23.9% and 7.1%, and the rates of conjunctival wound leak were 6.5% and 2.4% in Hh and control group respectively. There was no significant change in corneal endothelial density following the use of Hh (P > 0.05). CONCLUSION: The study indicates that Hh is a safe and effective antiproliferative agent for the use in glaucoma filtering surgery, it not only can increase the success probability considerably, but also maintain at least the postoperative IOP at relatively low normal level for 3 years. PMID- 11877228 TI - [Studies of mechanism of primary angle closure glaucoma using ultrasound biomicroscope]. AB - OBJECTIVE: To investigate the mechanism of primary angle closure glaucoma (PACG) especially the role of the changes of ciliary body using ultrasound biomicroscope (UBM) combined with general A scan. METHOD: 99 eyes with PACG, including 50 eyes with acute and 49 eyes with chronic PACG, were evaluated and 58 normal eyes were in the control group. RESULTS: Compared with the normal eyes the PACG eyes had their anatomic features. They had a smaller corneal diameter, shorter axial length, shallower anterior chamber, thicker lens, more relative anterior location of lens, swelling of ciliary process and anterior rotation of ciliary body. All these differences were significant statistically (P < 0.05). Compared with the chronic PACG eyes the acute PACG eyes had a shallower anterior chamber and a more anteriorly situated lens position. The differences were significant statistically (P < 0.01). Compared with the normal eyes the acute PACG eyes had a shorter ciliary body - lens distance and the difference was significant (P < 0.05); the chronic PACG eyes also had a shorter ciliary body-lens distance, but the difference was not significant (P > 0.05). CONCLUSIONS: The PACG eyes have their anatomic features. Among these factors the pupillary block caused by the anterior displacement of lens and the narrow angle induced consequently are the common mechanism. The swelling ciliary process, the positional changes of the ciliary body and age-related thicker lens are possibly the initial causes causing the forward movement of lens, and the pupillary block plays a more important role in the pathogenesis of acute PACG. PMID- 11877229 TI - [Quantitative real time measurement of iris configuration in living human eyes]. AB - OBJECTIVE: To evaluate the intraobserver and interobserver reproducibility of real time measurement of iris morphology in living human eyes. METHODS: Based on the platform of software (Autocad, version 12), we developed an ultrasound biomicroscopy (UBM) image assistant measuring system. Using the system, we can perform the iris configuration quantitative measurement in living eyes. The measuring parameters include: iris rest length, radius of iris curvature and the thickness of different part of iris. Ten anterior segment images of one normal individual were obtained by a single operator to evaluate the intraobserver reproducibility of image capture, and ten times of measurement of one image were performed by a single operator to assess the reproducibility of image measurement. The measurements of three independent observers were compared to investigate interobserver reproducibility in quantitative measurement. Intraobserver and interobserver reproducibility of measurement were assessed by calculating the coefficient of variation for each individual observer and by using F test to detect a difference among the observers. The iris configurations of 96 subjects (192 eyes) were measured. RESULTS: Intraobserver reproducibility ranged from 0.9 - 4.9% in all measured parameters. Interobserver reproducibility for some of the measured parameters varied considerably and was affect by subjective interpretation of visualized anatomic landmarks. The preliminary measured parameters in Chinese show that iris rest length is 3.699 +/- 0.397 mm, radius of iris curvature is 9.101 plus minus 1.408 mm, the average thickness of iris is 0.406 +/- 0.042 mm. CONCLUSIONS: The intraobserver reproducibility and the measured accuracy can fit the requisition of the ocular biometry, physiology and pharmacology studies. The method supplies a new assistance for UBM image measurement. PMID- 11877230 TI - [The effect of elevated hydrostatic pressure on expression level of type III collagen mRNA by lamina cribrosa cells in vitro]. AB - OBJECTIVE: To investigate the effect of elevated hydrostatic pressure on the expression level of type III collagen mRNA by lamina cribrosa cells (LCCs) and approach the action of changes of extracellular stroma on glaucomatous optic nerve damage. METHODS: LCCs were cultured and submitted to elevated hydrostatic pressure (4.67 - 5.33 kPa). We observed collagen type III mRNA level by dot-blot hybridization and computer imaging analysis. RESULTS: Under the pressure for 3 days, the amounts of collagen III mRNA were 0.5828 +/- 0.0517 in optical density (OD), compared with the same period controls (0.2832 +/- 0.0613), the difference was statistically significant (P < 0.05). When exposed to pressure up to 6 days, the collagen III mRNA level remained somewhat higher than that of the control, but there was no significant difference (P > 0.05). After 9 to 12 days of exposure to pressure, the mRNA amounts were decreased respectively to 0.3246 +/- 0.0957 and 0.2164 +/- 0.0171 in OD that were lower than that of the control in the respective same periods, and the differences between the experimental and the control had no significance (P > 0.05). CONCLUSION: Elevated hydrostatic pressure may increase expression level of collagen III mRNA by LCCs in a certain period. PMID- 11877231 TI - [Biochemical analysis of proteoglycan in bovine trabecular meshwork and its significance]. AB - OBJECTIVE: To determine the kinds of proteoglycan (PG) monochain and their distribution in the cell layer and cultural medium of bovine trabecular meshwork (BTM) culture. METHODS: (3)H-glycosamine was incorporated in BTM cells in the third passage. PGs were extracted by 4 mol/L guanidine hydrochloride. The aliquots were chromatographed on Sephadex G-50 DEAE-Sephacel, Sepharose 4B and Sepharose CL-6B gel respectively. Glycosaminoglycan was obtained by basic hydrolysis and enzyme degradation, and scintillation counting was used for the assay of radioactivity. RESULTS: Four different PGs were analyzed. The composition of PGs in cell layer was hyaluronic acid PG 30.2%, heparin sulfate PG 20.9%, chondroitin sulfate PG 21.6%, dermatan sulfate PG 18.1% and others 9.2%, while in the medium was 24.2%, 20.1%, 25.7%, 23% and 7%, respectively. CONCLUSIONS: PGs are confirmed to be present in the extracellular matrix of BTM cells in vitro. BTM cells also have the ability to secrete these macromolecules into the medium. Abnormal synthesis, secretion, accumulation and changes of PG pattern in physiologic conditions and under any drugs may contribute to the pathogenesis and treatment of primary open-angle glaucoma. PMID- 11877233 TI - [Accuracy and predictability of photorefractive keratectomy for astigmatism correction]. AB - OBJECTIVE: To study the accuracy and predictability of photorefractive keratectomy (PKR) for astigmatism correction. METHODS: 53 eyes of 30 cases with compound myopia astigmatism and 33 eyes of 23 cases with simple myopia were treated with PRK and followed up for 6 months. The differences of corneal refractive power provided by corneal topography between the values at postoperative 6 months and that of the preoperative one were calculated with Holladay Ways to get real corrective degree and axis of astigmatism. The results were analyzed and compared with the preoperative predicted values. RESULTS: Six months after PRK, among the compound myopia astigmatism group, the ratio of over correction or under correction of real corrected astigmatic power within +/- 0.50D was 60.4%, the ratio of axis deviation between real and predicted correction within +/- 15 degrees was 58.5%. Among simple myopia group, 48.5% appeared more than -0.50 D myopia astigmatism in real correction, and the cylinder axis position was within 180 degrees +/- 44 degrees. The ratios of uncorrected visual acuity after PRK in the two groups equal to or better than 1.0 were 77.4% and 78.8%, respectively. There was no difference between these two groups (chi(2) = 0.024, P > 0.75). CONCLUSIONS: The method of myopia astigmatism correction using this PRK machine needs to be improved. After PRK treatment, the visual acuity of myopia astigmatism is similar to that of simple myopia group. It shows good efficacy. It is indicated that for myopia astigmatism lower than 2.00D, especially for the astigmatism with the rule, PRK could be performed according to equivalent spherical value. PMID- 11877232 TI - [The regulating effects of dopamine receptors on the level of adenosine 3, 5 cyclic monophosphate in bovine trabecular cells in vitro]. AB - OBJECTIVE: To assay the regulating effects of dopaminergic agonists and antagonists on adenosine 3, 5-cyclic monophosphate (cAMP) level in bovine trabecular cells (BTC) in vitro. METHOD: H(3)-cAMP was used in protein binding assay to determine the level of cAMP in bovine BTC. RESULTS: Dopamine, a dopamine receptor agonist, and fenoldopam, a selective DA(1) receptor agonist, caused a fold increase of cAMP in cultured BTC in vitro. These effects were blocked by halporide, a non-selective dopamine antagonist, and SCH23309, a selective DA(1) antagonist, but not blocked by phenoxybenzamin, an alpha-adrenergic antagonist, and timolol, a beta-adrenergic antagonist. CONCLUSION: Bovine trabecular cells contain not only beta-receptors but also DA(1) receptors. PMID- 11877234 TI - [Correlations between factors and intraocular pressure following photorefractive keratectomy]. AB - OBJECTIVE: To evaluate the factors related to intraocular pressure (IOP) after excimer laser photorefractive keratectomy (PRK). METHODS: 86 cases 150 eyes of myopia followed over six months after PRK were studied with noncontact tonometer (NCT). RESULT: After PRK, a decrease in the IOP was observed. Post-operatively, the IOP reading at 1st month was the highest. The reduction of IOP may be related to the decrease in corneal thickness and corneal curvature (r = 0.361, P < 0.01; r = 0.188, P < 0.05). Regression equation for IOP drop (kPa) = - 0.059 - 0.038X(1) + 0.009X(2)[X(1) = ablation depth (microm), X(2) = decrease in corneal curvature (D)]. CONCLUSION: IOP readings after PRK are influenced by corneal thickness, corneal curvature and eye drug times. PMID- 11877235 TI - [Reconstructive lacrimal gland with free submandibular gland transfer for management of xerophthalmia]. AB - OBJECTIVE: In order to seek more efficient surgical method to treat total xerophthalmia. METHOD: Microvascular submandibular gland transfer was used to reconstruct lacrimal gland for management of the dry eye. 5 cases (6 eyes) with total xerophthalmia were operated with this method. RESULTS: All the patients were followed up for 1 - 30 months. The results were satisfactory, the submandibular salivary gland relieved the dryness and improved visual acuities with negligible side effect. CONCLUSION: Microvascular submandibular gland transfer is potentially the best surgical solution to total xerophthalmia. PMID- 11877236 TI - [Limbal epithelial autograft transplantation for treatment of pterygium]. AB - OBJECTIVE: To observe the therapeutic effects of limbal epithelial autograft transplantation for treatment of pterygium. METHODS: Limbal epithelial autograft transplantation was performed on 68 cases (76 eyes) with pterygium or its recurrent lesion. The post-operative follow-up periods ranged from 6 to 18 months (mean, 9.8 months). RESULTS: Of 68 cases (76 eyes), there were 56 cases (62 eyes) with stable epithelial healing, recovery of corneal transparency and no abnormal proliferation of pterygium-like tissue, and 12 cases (14 eyes) loss of follow-up. CONCLUSION: To provide new stem cell source for injured limbus with limbal epithelial autograft transplantation is a reasonable therapeutic method for treatment of pterygium. PMID- 11877237 TI - [Clinical analysis of primary orbital varix]. AB - OBJECTIVE: To analyze and summarize the clinical and imaging signs, diagnosis and treatment of primary orbital varix. METHOD: 218 cases of primary orbital varix in 20 years were analyzed. RESULTS: Positional proptosis was the primary sign. Ultrasound examination showed that a characteristic well-out-lined lesion with low or without reflection of sound appeared in the orbital fat when the internal carotid vein was compressed. CONCLUSION: To recognize the characteristic imaging signs of orbital varix in ultrasound examination is valuable for the choice of treatment. PMID- 11877238 TI - [A study of cytokines in pathogenesis of thyroid-related immune orbitopathy]. AB - OBJECTIVE: To explore the pathogenesis of thyroid-related immune orbitopathy (TRIO). METHODS: Human orbital fibroblasts were cultured in vitro and immunohistochemical techniques were used to study cytokines that induce effect on HLA-II antigen expression of orbital fibroblasts. RESULTS: It is shown that interferon-gamma (IFN-gamma), interleukin-1 (IL-1) and tumor necrosis factor (TNF) could induce the expression of orbital fibroblast HLA-II antigens. The induced effect of IFN-gamma is of good linear correlations with time and concentration. CONCLUSION: HLA-II antigen expression can be induced by IFN-gamma, IL-1 and TNF. In the course of immune reaction and pathogenesis of TRIO, cytokines play important roles. PMID- 11877239 TI - [Secondary surgical management of massive suprachoroidal hemorrhage]. AB - OBJECTIVE: To evaluate the surgical techniques and the efficacy for massive suprachoroidal hemorrhage (MSH). METHODS: Secondary surgery performed on 11 cases of MSH occurring during or after intraocular surgery was delayed for 11 to 28 days (mean, 15.4 days). All eyes underwent posterior drainage sclerotomies under constantly maintained limbal fluid line pressure, followed by pars plana infusion and vitreoretinal surgery. The perfluorocarbon liquid was used intraoperatively in 6 cases. RESULTS: The drainage of the choroidal hemorrhage was successful in all cases. The blood drained from suprachoroidal space was completely liquified and chocolate in color. Tractional retinal detachment occurred in 2 eyes; 9 eyes had retinas normal in position. The mean follow-up was 7.8 months. Visual acuities were improved, >or=0.1 in 6 eyes. CONCLUSION: Immediate management of MSH includes watertight wound closure and medical treatment for elevated intraocular pressure, and secondary surgery was performed timely, including external drainage by creating sclerotomies and vitreoretinal surgery. The above methods of treatment have certain advantages and are promising for the management of MSH. PMID- 11877240 TI - [Relaxing peripheral retinotomy with endolaser in treatment of retinal detachment with severe anterior proliferative vitreoretinopathy]. AB - OBJECTIVE: To investigate the effects of relaxing peripheral retinotomy with endolaser for treatment of complicated retinal detachment with severe anterior proliferative vitreoretinopathy. METHODS: 44 eyes of complicated retinal detachment with severe anterior proliferative vitreoretinopathy underwent relaxing peripheral retinotomy with endolaser treatment and silicone oil injection. Outcome measures included retinal reattachment, visual acuity, intraocular pressure as well as these after silicone oil removal. RESULTS: After minimum follow-up of 3 months, the retina was completely attached in 44 eyes (8 of them with folds), 38 eyes achieved improvement of visual acuity. The silicone oil was removed in 22 eyes, the retina kept attached in 21 eyes of them. Hypotony (intraocular pressure < 10 mmHg) was seen in 8 of the 22 eyes with reattached retina. CONCLUSION: Relaxing peripheral retinopathy with endolaser in the surgery for complicated retinal detachment with severe anterior proliferative vitreoretinopathy is an effective procedure. Subretinal membrane peeling benefits a complete retinal reattachment. Hypotony occurred in some cases, therefore indications should be limited. PMID- 11877241 TI - [Retinal detachment after intraocular lens implantation]. AB - OBJECTIVE: To study the specificity of retinal detachment after intraocular lens implantation. METHODS: The specialty of this type of retinal detachment through the clinical observation of 44 eyes of 42 cases from 1991 to 1996 was analyzed. RESULTS: The retinal detachment after intraocular lens implantation is more severe than others. It is more difficult to find a hole or holes because posterior capsular opacification and the implanted intraocular lens interfere with the careful search. The retinal detachment may develop into a complicated one. Its cure rate was 80.0% in the immediate postoperative period. CONCLUSION: The retinal injury during intraocular lens implantation should be avoided. As the retinal detachment occurs after the implantation, a more careful search and a earlier scleral buckling or/and vitrectomy should be done. PMID- 11877242 TI - [Vitrectomy for treatment of retinal detachment caused by macular hole]. AB - OBJECTIVE: To evaluate the effect of vitrectomy and autoserum for treatment of retinal detachment caused by macular hole. METHODS: 19 eyes with retinal detachment caused by macular hole were treated with gas tamponade (C(3)F(8)). The follow-up periods ranged from 6 to 24 months. In the case whose macular hole failed to heal, removal of the remains of prefoveal membrane and/or scratching at the retinal pigment epithelium (RPE) and placing autoserum for 7 minutes in the macular hole were performed. RESULTS: Closure of macular hole was obtained in 15 eyes (80%). The postoperative visual acuity was 0.05 - 0.4. Macular hole failed to heal in 4 eyes. In one eye the remains of prefoveal membrane were removed; in another 3 eyes, in the macular hole scratching at the retinal pigment epithelium (RPE) and placing autoserum were performed and C(3)F(8) tamponade was used. Anatomical cure was obtained in all the 4 eyes. The final visual acuity was 0.1 or better. CONCLUSION: The findings show that removal of prefoveal membrane is imperative. Scratch at RPE with placement of autoserum in the macular hole is effective for the macular hole which has failed to respond to conventional vitrectomy with air tamponade. PMID- 11877243 TI - [A study of Na(+)-K(+)-ATP ase activity in erythrocyte membrane from diabetic retinopathy patients]. AB - OBJECTIVE: To study the changes of Na(+)-K(+)-ATPase activity in erythrocyte membrane and its significance. METHODS: Na(+)-K(+)-ATPase activity in erythrocyte membrane was investigated with malachite green colorimetric analysis in 40 patients with diabetic retinopathy (DR), 20 diabetic patients with non DR (NDR), and 20 sex and age metched normal control subjects. The relations of Na(+)-K(+) ATPase activity with content of Na(+) and K(+) erythrocyte deformability index (EDI), levels of fasting blood glucose (FBG), glycosylated hemoglobin (HbA(1)c), and the course of disease were also studied. RESULTS: Na(+)-K(+)-ATPase activity in erythrocyte membrane was significantly decreased in diabetic patients (P < 0.01). The decrease was most significantly in patients with DR (P < 0.01). The decreased Na(+)-K(+)-ATPase activity was related to the content of Na(+) and K(+) in erythrocyte, the changes of EDI, FBG, HbA(1)c, and the course of disease (P < 0.01). CONCLUSIONS: The decreased Na(+)-K(+)-ATPase activity in erythrocyte membrane affects the deformability of erythrocyte. It may contribute to the occurrence and development of DR. PMID- 11877244 TI - [A preliminary investigation on causes of failure of vitreoretinal surgery for complicated rhegmatogenic retinal detachments]. AB - OBJECTIVE: To evaluate the failure causes of vitreoretinal surgery for complicated rhegmatogenic retinal detachment. METHODS: Vitreoretinal (VR) surgery was performed on 477 cases (479 eyes) with complicated rhegmatogenic retinal detachment, the operative procedures including vitrectomy, peeling of pre-retinal membrane, fluid-air exchange, inert gas (SF(6) or C(3)F(8)) or silicone tamponade. RESULTS: On discharge, the operation was effective in 349 eyes (72.87%) and failed in 130 (27.13%) eyes. CONCLUSION: The multiple stepwise multivariate regression shows that the factors significantly influencing VR surgery are silicone tamponade, giant retinal tears, SF(6) tamponade, iatrogenic tear, anterior proliferative vitreoretinopathy (PVR), subretinal proliferation, PVR, C(3)F(8) tamponade, operative frequency and choroidal detachments. PMID- 11877245 TI - [Phacoemulsification and foldable lens implantation through a temporal clear corneal tunnel incision]. AB - OBJECTIVE: To investigate the therapeutic effects, advantages and disadvantages of phacoemulsification and foldable lens implantation through a temporal clear corneal incision. METHODS: Eight hundreds and twelve patients (845 eyes) underwent phacoemulsification and foldable lens implantation through a temporal clear corneal tunnel incision which has a groove of 0.30 mm deep, a tunnel length of 1.75 mm, an internal opening of 3.20 mm and an external opening of 3.50 mm. The postoperative uncorrected visual acuity, the changes of corneal topography, astigmatism and endothelia were observed. RESULTS: The proportions of uncorrected visual acuity of 0.5 or better were 61.30% and 79.05% respectively at one day and a month after the operation. Postoperatively, topographical changes displayed temporal flattening near the incision in the early stage. The change of central corneal astigmatism was mild. Endothelial cell loss was 8.23%. The complications included burn of the incision (0.40%), tear of the posterior capsule (1.07%), subluxation of the lens (0.12%), endophthalmitis (0.12%) and cystoid macular edema (0.24%). CONCLUSION: The phacoemulsification and foldable lens implantation through a temporal clear corneal incision is convenient and effective. PMID- 11877246 TI - [An analysis of treatment of intravitreal lens fragments following phacoemulsification]. AB - OBJECTIVE: To observe and evaluate the outcome of vitrectomy in the treatment of dropped lens fragments. METHOD: Retrospectively the authors reviewed and analyzed 58 cases of dropped lens fragments that were treated with vitrectomy from October 1992 to October 1995 in Royal Victoria Eye and Ear Hospital, Melbourne, Australia and followed for at least 3 months. RESULTS: At the last follow-up, the remained complications were cystoid macular edema in three cases (5.2%), retinal detachment in two cases (3.4%) and hypotony in one case (1.7%). Other complications were all resolved. The final vision was significantly better than that before vitrectomy. There were 38 eyes (65.5%) with final visual acuities of 0.5 or better, and only 6 eyes (6.9%) with < 0.05. CONCLUSION: Once the lens fragments are dropped into the vitreous during phacoemulsification, the ocular anterior segment should be properly managed, if possible intraocular lens is inserted, and then vitrectomy should be performed as soon as possible. Generally, with the above method good therapeutic effects can be obtained. PMID- 11877247 TI - [Temporary keratoprosthesis for surgery of severe injuries of ocular anterior and posterior segments]. AB - OBJECTIVE: To evaluate the treatment of severe ocular injuries of anterior and posterior segments with temporary keratoprosthesis used in combined operations. METHODS: With temporary keratoprosthesis a combined operation including pars plana vitrectomy, air-fluid exchange, silicone oil fill in, use of perfluorocarbon liquid, foreign body removal and partial penetrating keratoplasty was performed on 18 eyes of 18 patients with severe ocular injuries, and the postoperative changes and recovery were observed. RESULTS: All eyes were rescued, the visual acuity was increased in 12 eyes, above 0.1 being in 5 eyes, retinal detachment recurred in 3 of 10 eyes, vitreous opacity was present in 1 eye, secondary glaucoma in 1 eye, and atrophy in 1 eye as its preoperative status. CONCLUSION: Temporary keratoprosthesis used in the combined surgeries of anterior and posterior ocular segments is an effective method for treatment of severe ocular injuries. PMID- 11877248 TI - [Application of ultrasound microscope for the reattachment surgery of detachment of ciliary body]. AB - OBJECTIVE: To evaluate the advantages of ultrasound biomicroscope (UBM) used for the reattachment surgery of traumatic ciliary body detachment. METHODS: 33 cases diagnosed by UBM as ciliary body detachment were studied. The surgical results of these cases were analyzed and compared with that of 83 cases which were simply diagnosed by a gonioscope. RESULTS: 31 of the 33 cases regained the normal intraocular pressure (IOP) after once of operation (94%), and all cases achieved successful results after twice of operation (100%). Of the 83 cases examined only by gonioscopy, the rates of reaching normal IOP after the 1(st), 2(nd) and 3(rd) operations were 82%, 94% and 96% respectively. CONCLUSIONS: These results suggest that the UBM be useful for the tiny and narrow detachment of ciliary body which can not be seen by a gonioscope. UBM has important clinical value in diagnosis, guide to the range of suture and follow-up observation. With the use of UBM, the frequency of operation necessary for surgical success and blindness in action in the operation will be reduced, and the successful rate of once of operation will be increased significantly. PMID- 11877249 TI - [The diagnostic value of high field MRI for choroidal melanoma]. AB - OBJECTIVE: To clarify the characteristic magnetic resonance imaging (MRI) features of choroidal melanomas by the high field MRI to improve the accuracy of diagnosis and localization and to differentiate choroidal melanomas from other choroidal lesions. METHODS: 12 patients with choroidal melanomas were examined by GE 1.5T magnet and heat coil, fat suppression technique and GD-DPTA as contrast agent were also used. Ten patients underwent operations and the histopathologic diagnosis was confirmed. RESULTS: The melanomas of the 12 patients had relatively high signals on T(1) weighted image and relatively low signals on T(2) weighted image. This is because of the paramagnetic properties of melanin. The melanomas with melanin can produce stable free radicals, both T(1) and T(2) relaxation times are shortened, with this we can differentiate melanomas from other choroidal lesions. CONCLUSION: Sagittal, coronal, axial and oblique scanning with small feild of view for the orbit by MRI can be made to show the exact position of melanomas and differentiate them from other choroidal lesions. PMID- 11877250 TI - [A clinical study on reducing immunologic rejection in combining corneal lamellar autografting with allografting]. AB - OBJECTIVE: To reduce the rate of immunologic rejection, corneal lamellar autografting with allografting were combined, making immune cells less attack on allograft. METHODS: The pterygium having encroached upon the pupillary area was removed surgically, and the combination of autograft with allograft was transplanted on the peripheral and central cornea in 23 cases (24 eyes). The control group of 13 cases (13 eyes) was established in which the pterygium and dermoids were removed, and lamellar allografts were transplanted. The rate of rejection and their effects were compared. RESULTS: The rate of transparent autograft and allograft was 100%, no rejection and recurrence of pterygium occurred in any case in the combined group, while in the control group, the rejection rate was 30.8% and recurrence rate, 20.0%. CONCLUSION: The combination of lamellar autografting with allografting can reduce the rate of rejection and recurrence of pterygium. PMID- 11877251 TI - [A research on clinical values of corneal topography and keratometer in measuring astigmatism]. AB - OBJECTIVE: To compare the accuracy of corneal topography, keratometer and cycloplegic retinoscopy in measuring astigmatism for refractive surgery. METHODS: Three hundred and sixty eyes with ametropia were randomly selected. The results of corneal topography, that of keratometer and cycloplegic retinoscopy in measuring astigmatism before refractive surgery were compared, and the measuring results of corneal topography and that of keratometer for 72 eyes after photorefractive keratectomy (PRK) and 95 eyes after laser in situ keratomileusis (LASIK) were also compared. RESULTS: No significant difference was found in axes of astigmatism among the three groups. As for diopters of astigmatism, there was no significant difference between the group of corneal topography (DeltaSim K) and the group of keratometer (DeltaK), but we found significant difference between the group of cycloplegic retinoscopy and the other two groups before and after refractive surgery. Significant difference was also found between DeltaSimK and DeltaK after PRK and LASIK. CONCLUSIONS: Our investigation demonstrates that before refractive surgery the keratometer has a similar clinical value as that of corneal topography, and the DeltaSim K value is related to the graphic pattern of corneal topography. However, keratometer is limited in measuring mild changes in corneal curvature after refractive surgery, while the astigmatism expressed by DeltaSim K in corneal topography is more accurate. PMID- 11877252 TI - [Myopic regression after photorefractive keratectomy]. AB - OBJECTIVE: To investigate myopic regression after excimer laser photorefractive keratectomy (PRK) in patients with myopia. METHODS: 103 eyes with regression (>or= -1.00 D) found in the follow-up for more than 1 year were included in this study. The preoperative spherical equivalent was -4.00 - -14.50 D (-8.72 +/- 2.65 D). Patients were divided into two groups: (1) moderate myopia (-4.00 - -7.90 D) and (2) high myopia (>or= -8.00 D). RESULTS: A significant positive correlation was found between regression and the extent of attempted correction, the rate of regression in high myopia group was 6.93% and that in moderate group, 1.17%. The rate of regression in older age group was higher than that in younger cases. (P < 0.01). Relatively severe haze under the corneal epithelium often accompanies the regression, and decrease of the dosage of topical corticosteroid application or its withdrawal because of the development of corticosteroid ocular hypertention could lead to the regression. The stability of the refraction is not related to the difference in sex. The regression is often accompanied by the increase in K value. CONCLUSION: The regression is significantly related to the extent of attempted correction, patients' age, corneal haze and topical application of corticosteroid. PMID- 11877253 TI - [Effects of mitomycin C on haze after photorefractive keratectomy for myopia in rabbits]. AB - OBJECTIVE: To investigate the effects of mitomycin C on haze after photorefractive keratectomy. METHODS: Forty-five rabbits underwent bilateral 193 nm excimer laser photorefractive keratectomy to correct 8 diopters of myopia. All eyes were allocated randomly to be treated with 0.008% mitomycin C during operation, or 0.1% dexamethasone after operation, or no medical treatment as control. Clinical and histopathologic examinations were made with slit-lamp microscope, specular microscope, light microscope and transmission electron microscope. RESULTS: At 4, 8 weeks after operation, the corneal haze was significantly less in mitomycin C group than that in the control group or in dexamethasone group, and the corneal haze was significantly less in dexamethasone group than that in the control group. At 1, 4, 8 weeks, the number of keratocyte in the anterior stroma of ablation area was significantly less in mitomycin C group than in the control group or in dexamethasone group. The difference between dexamethasone group and the control group was insignificant. The changes in time of reepithelialization, thickness of epithelium and keratocyte number of anterior stroma in ablation area were statistically insignificant among all groups. CONCLUSION: Mitomycin C can reduce corneal haze by inhibiting the proliferation of keratocyte and has no toxicity on cornea. Mitomycin C is more effective than dexamethasone on haze. PMID- 11877256 TI - Myeloid cell factor-1 is a critical survival factor for multiple myeloma. AB - Multiple myeloma (MM) is characterized by the accumulation of malignant plasma cells in the bone marrow caused primarily by failure of normal homeostatic mechanisms to prevent the expansion of postgerminal center plasma cells. We have examined the molecular mechanisms that promote the survival of MM cells and have identified a key role for myeloid cell factor-1 (Mcl-1), an antiapoptotic member of the Bcl-2 family. These experiments were initiated by the observation that MM cells were exquisitely sensitive to culture in the presence of actinomycin D: caspase activation occurred within 3 hours of treatment and cells were not protected by interleukin-6, the main MM cell growth and survival factor. Actinomycin D-induced apoptosis was blocked by proteasome inhibitors, suggesting that a labile protein was required for MM cell survival. Further analysis demonstrated that Mcl-1 was likely to be the labile factor governing MM cell survival. Mcl-1 protein levels decreased rapidly after culture in the presence of actinomycin D in concordance with effector caspase activation, but addition of proteasome inhibitors reversed the loss of Mcl-1 and maintained cell viability. The levels of other antiapoptotic proteins, including Bcl-2 and members of the inhibitors-of-apoptosis family, were unaffected by these interventions. Furthermore, Mcl-1 antisense oligonucleotides caused a rapid down-regulation of Mcl-1 protein levels and the coincident induction of apoptosis, whereas overexpression of Mcl-1 delayed actinomycin D-induced apoptosis with kinetics that correlated with expression levels of Mcl-1. These data indicate that Mcl-1 expression is required for the survival of MM cells and may represent an important target for future therapeutics. PMID- 11877257 TI - Processing of the lipocalin alpha(1)-microglobulin by hemoglobin induces heme binding and heme-degradation properties. AB - Alpha(1)-microglobulin is a 26-kd protein, widespread in plasma and tissues and well-conserved among vertebrates. Alpha(1)-microglobulin belongs to the lipocalins, a protein superfamily with highly conserved 3-dimensional structures, forming an internal ligand binding pocket. The protein, isolated from urine, has a heterogeneous yellow-brown chromophore bound covalently to amino acid side groups around the entrance of the lipocalin pocket. Alpha(1)-microglobulin is found in blood both in free form and complex-bound to immunoglobulin A (IgA) via a half-cystine residue at position 34. It is shown here that an alpha(1) microglobulin species, which we name t-alpha(1)-microglobulin (t = truncated), with a free Cys34 thiol group, lacking its C-terminal tetrapeptide, LIPR, and with a more polar environment around the entrance of the lipocalin pocket, is released from IgA-alpha(1)-microglobulin as well as from free alpha(1) microglobulin when exposed to the cytosolic side of erythrocyte membranes or to purified oxyhemoglobin. The processed t-alpha(1)-microglobulin binds heme and the alpha(1)-microglobulin-heme complex shows a time-dependent spectral rearrangement, suggestive of degradation of heme concomitantly with formation of a heterogeneous chromophore associated with the protein. The processed t-alpha(1) microglobulin is found in normal and pathologic human urine, indicating that the cleavage process occurs in vivo. The results suggest that alpha(1)-microglobulin is involved in extracellular heme catabolism. PMID- 11877258 TI - Successful treatment of murine beta-thalassemia intermedia by transfer of the human beta-globin gene. AB - The beta-thalassemias are caused by more than 200 mutations that reduce or abolish beta-globin production. The severity of the resulting anemia can lead to lifelong transfusion dependency. A genetic treatment based on globin gene transfer would require that transgene expression be erythroid specific, elevated, and sustained over time. We report here that long-term synthesis of chimeric hemoglobin (mualpha(2):hubeta(A)(2)) could be achieved in mice with beta thalassemia intermedia following engraftment with bone marrow cells transduced with a lentiviral vector encoding the human beta-globin gene. In the absence of any posttransduction selection, the treated chimeras exhibit durably increased hemoglobin levels without diminution over 40 weeks. Ineffective erythropoiesis and extramedullary hematopoiesis (EMH) regress, as reflected by normalization of spleen size, architecture, hematopoietic colony formation, and disappearance of liver EMH. These findings establish that a sustained increase of 3 to 4 g/dL hemoglobin is sufficient to correct ineffective erythropoiesis. Hepatic iron accumulation is markedly decreased in 1-year-old chimeras, indicating persistent protection from secondary organ damage. These results demonstrate for the first time that viral-mediated globin gene transfer in hematopoietic stem cells effectively treats a severe hemoglobin disorder. PMID- 11877259 TI - Genetic pathways in therapy-related myelodysplasia and acute myeloid leukemia. AB - Therapy-related acute myeloid leukemia (t-AML) in most cases develops after chemotherapy of other malignancies and shows characteristic chromosome aberrations. Two general types of t-AML have previously been identified. One type is observed after therapy with alkylating agents and characteristically presents as therapy-related myelodysplasia with deletions or loss of the long arms of chromosomes 5 and 7 or loss of the whole chromosomes. The other type is observed after therapy with topoisomerase II inhibitors and characteristically presents as overt t-AML with recurrent balanced chromosome aberrations. Recent research suggests that these 2 general types of t-AML can now be subdivided into at least 8 genetic pathways with a different etiology and different biologic characteristics. PMID- 11877260 TI - BCR ligation reprograms B cells for migration to the T zone and B-cell follicle sequentially. AB - We have studied the impact of B-cell receptor (BCR) or CD40 ligation on the in vitro chemotactic response of tonsillar B cells to 4 chemokines: stromal cell derived factor (SDF)-1alpha, macrophage inflammatory protein (MIP)-3alpha, MIP 3beta, and B-cell-attracting chemokine (BCA)-1. In the tonsil, SDF-1 and MIP 3alpha are both expressed in the crypt epithelium, while MIP-3beta is found in the T zone and BCA-1 in the follicles. Resting virgin and memory B cells display a similar chemotaxis pattern, and they both have the potential to migrate in vitro to all 4 chemokines studied. This pattern of responsiveness is strongly modified by a surrogate antigen (Ag) but is not altered by CD40 ligand. We report here that surrogate Ag induces a profound and sustained suppression of the response to the crypt chemokines SDF-1alpha and MIP-3alpha, while it exacerbates the migratory response to MIP-3beta. The effect of surrogate Ag on the response to BCA-1 is biphasic: After an initial phase of suppression, chemotaxis toward BCA-1 is strongly up-regulated. Our results suggest that Ag is primarily responsible for reprogramming the B-cell chemotaxis responsiveness during the humoral response. We propose that it initiates an ordered change of the chemotaxis machinery allowing Ag-activated B cells to relocate in the T zone and B-cell follicles sequentially. PMID- 11877261 TI - Does treatment with intermittent infusions of intravenous anti-D allow a proportion of adults with recently diagnosed immune thrombocytopenic purpura to avoid splenectomy? AB - This study explored whether repeated infusions of intravenous anti-D could allow adults with recently diagnosed immune thrombocytopenic purpura (ITP) who had failed an initial steroid course to postpone and ultimately avoid splenectomy. Twenty-eight Rh(+), nonsplenectomized adults with ITP diagnosed within 1 to 11 months and platelet counts 30 x 10(9)/L (30 000/microL) or below were enrolled. Anti-D was infused whenever the platelet count decreased to 30 x 10(9)/L (30 000/microL) or below. "Response" was defined as a platelet increase of more than 20 x 10(9)/L (20 000/microL) to more than 30 x 10(9)/L (30 000/microL) within 7 days of treatment. Patients were a median 3.5 months from ITP diagnosis at enrollment and had received a median of 2 previous therapies, including prednisone in 26 of 28 cases. They were followed for a median 26 months. A total of 93% responded to their initial infusion of anti-D, and 68% repeatedly responded with counts maintained above 30 x 10(9)/L (30 000/microL) using anti-D alone. Currently, 12 (43%) of 28 patients have been off all treatment for more than 6 months without undergoing splenectomy, 6 maintaining counts above 100 x 10(9)/L (100 000/microL). Seven continue on treatment, 8 underwent splenectomy, and 1 was lost to follow-up at 10 months. One patient discontinued anti-D because of toxicity. Patients with platelet counts at least 14 x 10(9)/L (14 000/microL) at enrollment were more likely to discontinue treatment (P <.05). Anti-D was an effective maintenance treatment for two thirds of Rh(+), nonsplenectomized adults with ITP who had failed an initial steroid course. Intermittent infusions of intravenous anti-D allowed more than 40% of these adults to avoid splenectomy and to achieve stable platelet counts off all therapy, even after many months of treatment. Platelet count at study entry was the primary predictor of outcome. PMID- 11877262 TI - Imatinib induces durable hematologic and cytogenetic responses in patients with accelerated phase chronic myeloid leukemia: results of a phase 2 study. AB - Chronic myelogenous leukemia (CML) is caused by expression of the BCR-ABL tyrosine kinase oncogene, the product of the t(9;22) Philadelphia translocation. Patients with CML in accelerated phase have rapidly progressive disease and are characteristically unresponsive to existing therapies. Imatinib (formerly STI571) is a rationally developed, orally administered inhibitor of the Bcr-Abl kinase. A total of 235 CML patients were enrolled in this study, of whom 181 had a confirmed diagnosis of accelerated phase. Patients were treated with imatinib at 400 or 600 mg/d and were evaluated for hematologic and cytogenetic response, time to progression, survival, and toxicity. Imatinib induced hematologic response in 82% of patients and sustained hematologic responses lasting at least 4 weeks in 69% (complete in 34%). The rate of major cytogenetic response was 24% (complete in 17%). Estimated 12-month progression-free and overall survival rates were 59% and 74%, respectively. Nonhematologic toxicity was usually mild or moderate, and hematologic toxicity was manageable. In comparison to 400 mg, imatinib doses of 600 mg/d led to more cytogenetic responses (28% compared to 16%), longer duration of response (79% compared to 57% at 12 months), time to disease progression (67% compared to 44% at 12 months), and overall survival (78% compared to 65% at 12 months), with no clinically relevant increase in toxicity. Orally administered imatinib is an effective and well-tolerated treatment for patients with CML in accelerated phase. A daily dose of 600 mg is more effective than 400 mg, with similar toxicity. PMID- 11877263 TI - Incidence of venous thromboembolism in asymptomatic family members who are carriers of factor V Leiden: a prospective cohort study. AB - In a prospective cohort study, we assessed the incidence of spontaneous and risk period-related venous thromboembolism (VTE) in asymptomatic family members of patients who experienced VTE and had the factor V Leiden mutation. In all, 561 family members of 131 probands were included, 313 of whom were carriers (299 heterozygous and 14 homozygous) and 248 of whom were noncarriers of the factor V Leiden mutation. Average follow-up was 4 years (range, 4 months-6 years). There were 1255 and 984 observation-years of follow-up in carriers and noncarriers, respectively. Eight episodes of VTE occurred in heterozygous carriers, resulting in an annual incidence of 0.67% (95% confidence interval [CI], 0.29-1.33). Two events occurred in the absence of associated risk factors, determining an annual incidence of spontaneous VTE of 0.17% (95% CI, 0.02-0.6). Only one VTE (risk period-related) occurred in noncarriers, with an annual incidence of 0.1% (95% CI, 0.003-0.56). Relative risk for VTE in heterozygous carriers compared with noncarriers of the factor V Leiden mutation was 6.6 (95% CI, 1.1-39.8). Risk period-related VTE occurred with an incidence of 18% and 5% per risk period in heterozygous carriers and in noncarriers, respectively. Thus, the low rate of VTE in asymptomatic family members carrying the mutation did not justify continuous anticoagulant prophylaxis. Screening families of symptomatic probands with the factor V Leiden mutation has the potential to identify those asymptomatic carriers who might benefit from thromboprophylaxis during risk periods. PMID- 11877264 TI - Unrelated donor marrow transplantation for myelodysplastic syndromes: outcome analysis in 510 transplants facilitated by the National Marrow Donor Program. AB - Between April 1988 and July 1998, 510 patients with myelodysplastic syndromes (MDS) underwent unrelated donor bone marrow transplantation (BMT) facilitated by the National Marrow Donor Program. Median age was 38 years (range, <1-62 years). Several conditioning regimens and graft-versus-host disease (GVHD) prophylaxis methods were used, and T-cell depletion was used in 121 patients. Donors were serologically matched for HLA-A, -B, and -DRB1 antigens for 74% of patients. Of 437 patients evaluable for engraftment, 24 (5% cumulative incidence, with 95% confidence interval [CI] of 3%-7%) failed to engraft, and an additional 33 (8% cumulative incidence; 95% CI, 6%-10%) had late graft failure. Grades II to IV GVHD developed in 47% of patients (95% CI, 43%-49%), and limited and extensive chronic GVHD developed at 2 years in 27% (95% CI, 24%-30%). The incidence of relapse at 2 years was 14% (95% CI, 11%-17%). Greater relapse was independently associated with advanced MDS subtype and no acute GVHD. The estimated probability of disease-free survival (DFS) at 2 years was 29% (95% CI, 25%-33%). Improved DFS was independently associated with less advanced MDS subtype, higher cell dose, recipient cytomegalovirus (CMV) seronegativity, shorter interval from diagnosis to transplantation, and transplantation in recent years. Common causes of death were treatment-related complications accounting for 82% of fatalities. The 2-year cumulative incidence of treatment-related mortality (TRM) was 54% (95% CI, 53% 61%). Sixty-nine percent of TRM occurred within the first 100 days, and 93% occurred within the first year of transplantation. Higher TRM was independently associated with older recipient and donor age, HLA mismatch, and recipient CMV seropositivity. This study demonstrates that unrelated donor BMT cures a significant proportion of patients with MDS. TRM is the major problem limiting the success of unrelated donor BMT in MDS. The observations made in this study should facilitate the design of prospective trials aimed at improving the results of unrelated donor stem cell transplantation for MDS. PMID- 11877265 TI - Prognostic significance and modalities of flow cytometric minimal residual disease detection in childhood acute lymphoblastic leukemia. AB - Detection of minimal residual disease (MRD) in acute lymphoblastic leukemia (ALL) predicts outcome. Previous studies were invariably based on relative quantification and did not investigate sample-inherent parameters that influence test accuracy, which makes comparisons and clinical conclusions cumbersome. Hence, we conducted a prospective, population-based MRD study in 108 sequentially recruited children with ALL uniformly treated with the ALL-Berlin-Frankfurt Munster (ALL-BFM) 95 protocol in Austria (median follow-up of 40 months). Using sensitive, limited antibody panel flow cytometry applicable to 97% of patients, we investigated 329 bone marrow samples from 4 treatment time points. MRD was quantified by blast percentages among nucleated cells (NCs) and by absolute counts (per microliter). Covariables such as NC count, normal B cells, and an estimate of the test sensitivity were also recorded. Presence and distinct levels of MRD correlated with a high probability of early relapse at each of the time points studied. Sequential monitoring at day 33 and week 12 was most useful for predicting outcome independently from clinical risk groups: patients with persistent disease (> or =1 blast/microL) had a 100% probability of relapse, compared to 6% in all others. Absolute MRD quantification was more appropriate than relative, due to considerable variations in total NC counts between samples. Regeneration of normal immature B cells after periods of rest from treatment limited the test sensitivity. In conclusion, MRD detection by flow cytometry is a strong and independent outcome indicator in childhood ALL. Standardization regarding absolute quantification on the basis of NCs and assessment during periods of continuous treatment promise to increase the accuracy, simplicity, and cost efficiency of the approach. PMID- 11877266 TI - Clinicopathologic analysis of follicular lymphoma occurring in children. AB - Follicular lymphoma is a rare lymphoid malignancy in pediatric patients and consequently remains poorly characterized, particularly with respect to its immunophenotype and molecular pathogenesis. A total of 23 pediatric patients with follicular lymphoma were identified, with a median age of 11 years and a male-to female ratio of 2.3:1. Of the 19 patients for whom presenting clinical features were available, 15 patients had stage I, 1 had stage II, and 3 had stage III or IV disease. All tumors had a follicular architecture, and 74% of cases had grade 2 or 3 histologic features. All patients expressed CD20 and bcl-6, and 80% were positive for CD10. Bcl-2 expression was detected in only 5 of 16 cases. Consistent with this finding, bcl-2 gene rearrangements were detected in only 2 of 16 cases by polymerase chain reaction. These patients were treated primarily with cyclophosphamide, doxorubicin, vincristine, and prednisone-based chemotherapy; 4 patients also received involved-field irradiation. Of the 13 patients with available clinical follow-up, all but 2 achieved durable clinical remission. Importantly, all 4 patients with tumors diffusely positive for bcl-2 either presented with stage III/IV disease or had disease refractory to therapy, whereas patients with bcl-2-negative tumors uniformly had stage I disease, achieved complete remission, and experienced no relapses. These findings indicate that, in contrast to adult follicular lymphomas, dysregulated bcl-2 expression does not play a significant pathogenetic role in most pediatric follicular lymphomas. However, bcl-2 expression in pediatric follicular lymphoma identifies a subset of patients in whom disease is often disseminated at clinical presentation and is more refractory to combination chemotherapy. PMID- 11877267 TI - Randomized trial of different regimens of heparins and in vivo thrombin generation in acute deep vein thrombosis. AB - Low-molecular-weight and unfractionated heparins are frequently used to treat venous thromboembolism, but it is not known whether they are equally effective in inhibiting in vivo generation of thrombin. In this multicenter trial, 1048 patients were randomized to intravenous unfractionated heparin (group A), twice daily low-molecular-weight heparin (reviparin) for 1 week (group B), or once daily reviparin for 4 weeks (group C). All patients received vitamin K antagonists. Blood samples withdrawn at the baseline and at weeks 1 and 3 were analyzed using markers of in vivo thrombin generation and other coagulation parameters. During the first 3 weeks symptomatic recurrent deep vein thrombosis pulmonary embolism (DVT/PE) occurred in 17 (4.5%) of 375 patients in group A compared with 4 (1.0%) of 388 patients in group B, and 9 (2.4%) of 374 patients in group C. Forty percent of patients in group A, 53.4% in group B, and 53.5% in group C showed 30% or greater reduction in thrombus size assessed by venography. Patients in group B had significantly greater reduction in D-dimer, prothrombin fragments 1 and 2 (F1 + 2), endogenous thrombin potential (ETP), and thrombin antithrombin (TAT) complexes compared to groups A and C. Greater release of tissue factor pathway inhibitor (TFPI) and reduction in levels of thrombin activatable fibrinolysis inhibitor (TAFI) and fibrinogen were significantly more pronounced in group C patients. Reviparin administered twice daily plus vitamin K antagonist is more effective in inhibiting in vivo thrombin generation compared to intravenous unfractionated heparin plus vitamin K antagonist, and reviparin once daily produced significantly higher TFPI release and greater reduction in TAFI and fibrinogen levels. PMID- 11877268 TI - Allogeneic bone marrow transplantation for chronic myelogenous leukemia: comparative analysis of unrelated versus matched sibling donor transplantation. AB - Allogeneic bone marrow transplantation (BMT) offers the only curative therapy for chronic myelogenous leukemia. We compared prospectively collected results of 2464 unrelated donor (URD) transplantations with 450 HLA-identical, matched sibling donor (MSD) transplantations performed at collaborating National Marrow Donor Program institutions. A total of 63% of URDs were matched at HLA-A, -B, and at DRB1 alleles; all MSDs were genotypically identical at major histocompatibility loci. URD recipients were younger (median 36 vs 39, P =.001) than MSDs and underwent BMT later after diagnosis (median 17 [0-325 months] vs 7 [1-118 months], P =.001) and less often in chronic phase (CP) (67% vs 82%, P =.001). Multivariate analysis demonstrated a significantly increased risk of graft failure and acute graft versus host disease after URD BMT. The risk of hematologic relapse was low after either matched URD or MSD transplantations. We observed significantly though modestly poorer survival and disease-free survival (DFS) after URD transplantations. However, for those undergoing transplantation during CP within 1 year from diagnosis, 5-year DFS was similar or only slightly inferior after matched URD versus MSD transplantation (age < 30: URD 61% +/- 8% vs MSD 68% +/- 15%, P =.18; 30-40: URD 57% +/- 9% vs MSD 67% +/- 10%, P =.05; > 40: URD 46% +/- 9% vs MSD 57% +/- 9%, P =.02). Delay from diagnosis to BMT in CP patients led to substantially poorer 5-year DFS after matched URD than MSD BMT (CP 1-2 years: URD 39% +/- 6% vs MSD 63% +/- 12%; beyond 2 years: URD 33% +/- 7% vs MSD 50% +/- 20%). Outcome of matched URD BMT for early CP chronic myelogenous leukemia yields survival and DFS approaching that of MSD transplantation. However, delay may compromise URD outcomes to a greater extent. Improvements in URD and MSD transplantation are still needed, and results of newer, nontransplantation therapies should be evaluated against the established curative potential of URD and MSD marrow transplantation. PMID- 11877269 TI - Incidence and outcome of cytomegalovirus infections following nonmyeloablative compared with myeloablative allogeneic stem cell transplantation, a matched control study. AB - Nonmyeloablative allogeneic hematopoietic stem cell transplantation (HSCT) is increasingly being explored as therapy in patients who are not eligible for conventional myeloablative HSCT. Whether these transplants are associated with reduced risk of transplantation-related infections is unknown. We analyzed the incidence of posttransplantation cytomegalovirus (CMV) infections in 56 consecutive mycophenolate mofetil (MMF) patients with hematologic malignancies who underwent nonmyeloablative HSCT (TBI, 2Gy, day 0; MMF/cyclosporine after transplantation). In addition, 18 of 56 patients received 30 mg/m(2)/d fludarabine on days -4 to -2. Most donors were HLA matched and related (93%). Each case patient was matched to 2 controls who were treated by conventional HSCT during the same time period (January 1997 through April 2000). Matching criteria included CMV risk group, HSC source, donor type, age, and underlying diseases. No CMV disease occurred in the low (donor and recipient serologically negative) and intermediate (donor serologically positive and recipient negative) CMV risk groups during the first 100 days. Among cases at high risk for CMV (seropositive recipients), trends to less CMV antigenemia (P =.11), viremia (P =.16), and disease (P =.08) compared with controls were observed; all severe manifestations combined (CMV viremia and disease) were significantly reduced among cases (P =.01). However, by day 365, the overall incidence of CMV disease became similar in both groups. The onset of CMV disease was significantly delayed among case patients compared with controls (median, 130 days versus 52 days; P =.02). It was concluded that CMV disease was significantly delayed in nonmyeloablative cases, but that the overall 1-year incidence was similar to myeloablative HSCT patients. Therefore, nonmyeloablative HSCT patients should receive CMV surveillance beyond day 100 and pre-emptive ganciclovir treatment similar to that of myeloablative HSCT patients. PMID- 11877271 TI - Health-related quality of life of unrelated bone marrow donors in Japan. AB - To promote bone marrow donation, both the safety and well-being of healthy unrelated volunteer donors must be protected. This prospective cohort study evaluated donors' health-related quality of life (HRQOL) and identified factors associated with it. Using the Medical Outcomes Study Short Form 36 Health Survey (SF-36) before bone marrow harvesting (BMH), and again 1 week and 3 months after the donors' discharge, we evaluated HRQOL of 565 donors (329 men, 236 women) registered with the Japan Marrow Donor Program (JMDP). We also examined the data routinely collected by the JMDP, such as BMH-related problems and other demographic and medical variables, to determine whether such data could be used to predict donors' HRQOL after discharge. Mean scores of all pre-BMH SF-36 subscales showed better functioning than the national norm. One week after discharge, mean scores on physical functioning (PF) and role-physical (RP) subscales, indicative of physical states, and bodily pain (BP) were approximately 1 SD lower than the national norm; however, mental health (MH) and general health perception (GH) remained above normal; the most frequent BMH-related problems were pain at the donation site and lower back pain, which were associated with lower PF, RP, and BP scores. Female gender and duration of procedure predicted lower PF, RP, and BP. Three months after discharge, mean scores of all SF-36 subscales had returned to baseline levels. These data show that the adverse effects of BMH on donors' HRQOL are transient and can be minimized by better management of pain. PMID- 11877270 TI - A randomized comparison of native Escherichia coli asparaginase and polyethylene glycol conjugated asparaginase for treatment of children with newly diagnosed standard-risk acute lymphoblastic leukemia: a Children's Cancer Group study. AB - For this study, 118 children with standard-risk acute lymphoblastic leukemia (ALL) were given randomized assignments to receive native or pegylated Escherichia coli asparaginase as part of induction and 2 delayed intensification phases. Patients treated with pegaspargase had more rapid clearance of lymphoblasts from day 7 and day 14 bone marrow aspirates and more prolonged asparaginase activity than those treated with native asparaginase. In the first delayed intensification phase, 26% of native asparaginase patients had high-titer antibodies, whereas 2% of pegaspargase patients had those levels. High-titer antibodies were associated with low asparaginase activity in the native arm, but not in the pegaspargase arm. Adverse events, infections, and hospitalization were similar between arms. Event-free survival at 3 years was 82%. A population pharmacodynamic model using the nonlinear mixed effects model (NONMEM) program was developed that closely fit the measured enzyme activity and asparagine concentrations. Half-lives of asparaginase were 5.5 days and 26 hours for pegaspargase and native asparaginase, respectively. There was correlation between asparaginase enzymatic activity and depletion of asparagine or glutamine in serum. In cerebrospinal fluid asparagine, depletion was similar with both enzyme preparations. Intensive pegaspargase for newly diagnosed ALL should be tested further in a larger population. PMID- 11877273 TI - Epstein-Barr virus-specific human T lymphocytes expressing antitumor chimeric T cell receptors: potential for improved immunotherapy. AB - Primary T cells expressing chimeric receptors specific for tumor or viral antigens have considerable therapeutic potential. Unfortunately, their clinical value is limited by their rapid loss of function and failure to expand in vivo, presumably due to the lack of costimulator molecules on tumor cells and the inherent limitations of signaling exclusively through the chimeric receptor. Epstein-Barr virus (EBV) infection of B lymphocytes is near universal in humans and stimulates high levels of EBV-specific helper and cytotoxic T cells, which persist indefinitely. Our clinical studies have shown that EBV-specific T cells generated in vitro will expand, persist, and function for more than 6 years in vivo. We now report that EBV-specific (but not primary) T cells transduced with tumor-specific chimeric receptor genes can be expanded and maintained long-term in the presence of EBV-infected B cells. They recognize EBV-infected targets through their conventional T-cell receptor and tumor targets through their chimeric receptors. They efficiently lyse both. EBV-specific T cells expressing chimeric antitumor receptors may represent a new source of effector cells that would persist and function long-term after their transfer to cancer patients. PMID- 11877272 TI - Factors associated with outcome after unrelated marrow transplantation for treatment of acute lymphoblastic leukemia in children. AB - Acute lymphoblastic leukemia (ALL) is the most common indication for transplantation of marrow from unrelated donors in children. We analyzed results of this procedure in children with ALL treated according to a standard protocol to determine risk factors for outcome. From January 1987 to 1999, 88 consecutively seen patients with ALL who were younger than 18 years received a marrow transplant from an HLA-matched (n = 56) or partly matched (n = 32) unrelated donor during first complete remission (CR1; n = 10), second remission (CR2; n = 34), third remission (CR3; n = 10), or relapse (n = 34). Patients received cyclophosphamide and fractionated total-body irradiation as conditioning treatment and were given methotrexate and cyclosporine for graft-versus-host disease (GVHD) prophylaxis. Three-year rates of leukemia-free survival (LFS) according to phase of disease were 70% for CR1, 46% for CR2, 20% for CR3, and 9% for relapse (P <.0001). Three-year cumulative relapse rates were 10%, 33%, 20%, and 50%, respectively, and 3-year cumulative rates of death not due to relapse were 20%, 22%, 60%, and 41%, respectively, for patients with CR1, CR2, CR3, and relapse. Grades III to IV acute GVHD occurred in 43% of patients given HLA matched transplants and in 59% given partly matched transplants (P =.10); clinical extensive chronic GVHD occurred in 32% and 38%, respectively (P =.23). LFS rates were lower in patients with advanced disease (P <.0001), age 10 years or older (P =.002), or short duration of CR1 (P =.007). Thus, in addition to phase of disease, age and duration of CR1 were predictors of outcome after unrelated-donor transplantation for treatment of ALL in children. Outcome was particularly favorable in younger patients with early phases of the disease. PMID- 11877274 TI - Infant hypervitaminosis A causes severe anemia and thrombocytopenia: evidence of a retinol-dependent bone marrow cell growth inhibition. AB - Vitamin A is a pivotal biochemical factor required for normal proliferation and differentiation as well as for specialized functions, such as vision. The dietary intake of 1500 IU/day is recommended in the first year of life. Here, we report the case of an infant who had been given 62 000 IU/day for 80 days. The infant showed several clinical signs of retinol intoxication, including severe anemia and thrombocytopenia. Bone marrow showed a remarkably reduced number of erythroid and megakaryocytic cells. The interruption of vitamin A treatment was immediately followed by clinical and biochemical recovery. To clarify whether the effects of retinol are due to a direct action on bone marrow cell proliferation, we investigated the activity of retinol (both the drug and the pure molecule) on the growth of K-562, a multipotent hematopoietic cell line, and on bone marrow mesenchymal stem cells. We observed that vitamin A strongly inhibited the proliferation of the cells at concentrations similar to those reached in vivo. Subsequent biochemical analyses of the cell cycle suggested that the effect was mediated by the up-regulation of cyclin-dependent kinase inhibitors, p21(Cip1) and p27(Kip1). These are the first findings to demonstrate that infant hypervitaminosis A causes a severe anemia and thrombocytopenia and that this is probably due to the direct effect of the molecule on the growth of all bone marrow cellular components. Our data also suggest potential bone marrow functional alterations after excessive vitamin A intake because of emerging social habits. PMID- 11877275 TI - Increased binding and defective migration across fibronectin of cycling hematopoietic progenitor cells. AB - Engraftment of hematopoietic progenitor cells has been shown to decrease during cell cycle transit. We studied cell cycle-associated changes in adhesion and migration of mitotically activated cord blood CD34+ cells. Migration toward medium conditioned by the stromal-derived factor-1-producing cell line MS-5 was studied in bovine serum albumin- and fibronectin (Fn)-coated transwells. Migration was reduced in cycling CD34+ cells and long-term culture-initiating cells (LTC-ICs) compared with their noncycling counterparts across Fn but not across bovine serum albumin. Conversely, Fn binding was higher in cycling CD34+ cells and LTC-ICs compared with noncycling progenitor cells, while adhesion of both subsets to bovine serum albumin was undetectable. The contribution of alpha4 and alpha5 integrins in mediating adhesion and migration of activated CD34+ cells onto Fn was analyzed by neutralization experiments. While alpha4-mediated Fn binding decreased during G(2)/M, alpha5 integrin-mediated adhesion increased during transit from G(0)/G(1) to S and G(2)/M phases. As for migration, the contribution of alpha4 integrin was similar in all phases, whereas alpha5 directed migration was lower in G(2)/M compared with G(0)/G(1) and S phases. Defective migration of cycling CD34+ cells was not due to differences in alpha5 integrin expression. In conclusion, chemotaxis across Fn is less efficient in cycling progenitor cells in correlation with an increased Fn binding capacity. In addition, alpha4 and alpha5 integrin functions are independently modulated during cell cycle transit. PMID- 11877276 TI - Expression of C/EBPbeta from the C/ebpalpha gene locus is sufficient for normal hematopoiesis in vivo. AB - CCAAT/enhancer-binding proteins (C/EBPs) are critical transcriptional regulators of differentiation of hematopoietic cells. Previous studies have shown that targeted disruption of the C/ebpalpha gene results in a lack of granulocytic differentiation with an arrest at the stage of immature myeloblasts. By using a gene replacement strategy in which C/EBPbeta was expressed from the C/ebpalpha gene locus of C/EBPalpha-null mice, we have evaluated the ability of C/EBPbeta to function for C/EBPalpha in directing differentiation along the granulocytic pathway. We show that the morphology and the differential cell counts of the bone marrow and peripheral blood cells from C/EBPbeta knockin mice are indistinguishable from those of their wild-type littermates, indicating that hematopoiesis occurs normally in these animals. Additionally, we analyzed expression of 21 myeloid-specific genes, including markers for distinct stages of granulocytic differentiation, and found no significant differences in their levels of expression in the bone marrow of C/EBPbeta knockin and wild-type mice. These results imply that C/EBPbeta can substitute for C/EBPalpha during hematopoiesis when expressed from the C/ebpalpha gene locus. PMID- 11877277 TI - Gene expression profiling identifies significant differences between the molecular phenotypes of bone marrow-derived and circulating human CD34+ hematopoietic stem cells. AB - CD34+ hematopoietic stem cells are used clinically to support cytotoxic therapy, and recent studies raised hope that they could even serve as a cellular source for nonhematopoietic tissue engineering. Here, we examined in 18 volunteers the gene expressions of 1185 genes in highly enriched bone marrow CD34+ (BM-CD34+) or granulocyte-colony-stimulating factor-mobilized peripheral blood CD34+ (PB-CD34+) cells by means of cDNA array technology to identify molecular causes underlying the functional differences between circulating and sedentary hematopoietic stem and progenitor cells. In total, 65 genes were significantly differentially expressed. Greater cell cycle and DNA synthesis activity of BM-CD34+ than PB CD34+ cells were reflected by the 2- to 5-fold higher expression of 9 genes involved in cell cycle progression, 11 genes regulating DNA synthesis, and cell cycle-initiating transcription factor E2F-1. Conversely, 9 other transcription factors, including the differentiation blocking GATA2 and N-myc, were expressed 2 to 3 times higher in PB-CD34+ cells than in BM-CD34+ cells. Expression of 5 apoptosis driving genes was also 2 to 3 times greater in PB-CD34+ cells, reflecting a higher apoptotic activity. In summary, our study provides a gene expression profile of primary human CD34+ hematopoietic cells of the blood and marrow. Our data molecularly confirm and explain the finding that CD34+ cells residing in the bone marrow cycle more rapidly, whereas circulating CD34+ cells consist of a higher number of quiescent stem and progenitor cells. Moreover, our data provide novel molecular insight into stem cell physiology. PMID- 11877278 TI - Platelet-derived growth factor inhibits basic fibroblast growth factor angiogenic properties in vitro and in vivo through its alpha receptor. AB - Basic fibroblast growth factor (bFGF) and platelet-derived growth factor-BB (PDGF BB) modulate vascular wall cell function in vitro and angiogenesis in vivo. The aim of the current study was to determine how bovine aorta endothelial cells (BAECs) respond to the simultaneous exposure to PDGF-BB and bFGF. It was found that bFGF-dependent BAEC migration, proliferation, and differentiation into tubelike structures on reconstituted extracellular matrix (Matrigel) were inhibited by PDGF-BB. The role played by PDGF receptor alpha (PDGF-Ralpha) was investigated by selective stimulation with PDGF-AA, by blocking PDGF-BB-binding to PDGF-Ralpha with neomycin, or by transfecting cells with dominant-negative forms of the receptors to selectively impair either PDGF-Ralpha or PDGF-Rbeta function. In all cases, PDGF-Ralpha impairment abolished the inhibitory effect of PDGF-BB on bFGF-directed BAEC migration. In addition, PDGF-Ralpha phosphorylation was increased in the presence of bFGF and PDGF, as compared to PDGF alone, whereas mitogen-activated protein kinase phosphorylation was decreased in the presence of PDGF-BB and bFGF compared with bFGF alone. In vivo experiments showed that PDGF-BB and PDGF-AA inhibited bFGF-induced angiogenesis in vivo in the chick embryo chorioallantoic membrane assay and that PDGF-BB inhibited bFGF-induced angiogenesis in Matrigel plugs injected subcutaneously in CD1 mice. Taken together these results show that PDGF inhibits the angiogenic properties of bFGF in vitro and in vivo, likely through PDGF-Ralpha stimulation. PMID- 11877279 TI - Thrombocytopenia after second exposure to abciximab is caused by antibodies that recognize abciximab-coated platelets. AB - Thrombocytopenia, often severe, occurs in 1% to 2% of patients given the fibrinogen receptor antagonist abciximab, a chimeric Fab fragment containing murine specificity-determining and human framework sequences. The cause of this complication has not yet been defined. Studies of 9 patients who developed profound thrombocytopenia (platelets <10 x 10(9)/L [10 000/microL]) within a few hours of being given abciximab a second time showed that each had a strong immunoglobulin G (IgG) antibody that recognized platelets sensitized with abciximab. Five patients also had IgM antibodies. IgG antibodies reactive with abciximab-coated platelets were also found in 77 (74%) of 104 healthy subjects. However, the patient antibodies could be distinguished from "normal" ones in 2 ways: (1) only the patient antibodies reacted preferentially with platelets sensitized with the intact monoclonal antibody 7E3 from which the murine sequences in abciximab are derived; and (2) the "normal" antibodies could be inhibited by Fab fragments derived from normal human IgG, whereas the patient antibodies were relatively resistant to this treatment. The findings suggest that antibodies from the patients are specific for murine sequences in abciximab and are capable of causing life-threatening thrombocytopenia upon injection of this drug. The antibodies commonly found in healthy subjects are specific for the papain cleavage site of any Fab fragments and, although they react with abciximab coated platelets, appear not to cause significant thrombocytopenia. It may be possible to identify patients at risk for developing thrombocytopenia if given abciximab by screening for antibodies that recognize 7E3-coated platelets. PMID- 11877281 TI - Shear-dependent morphology of von Willebrand factor bound to immobilized collagen. AB - We have developed an immunogold von Willebrand factor (VWF) detection method that permits almost complete coverage of individual VWF molecules, and by this unequivocal localization and morphologic analysis of collagen-bound VWF by atomic force microscopy (AFM). Perfusion of gel filtration-purified VWF in parallel plate perfusion chambers over glass coverslips coated with calf skin collagen, followed by AFM imaging in air, enabled us to assess possible morphologic differences between VWF bound at low (0.07 N/m(2) = 0.7 dynes/cm(2)) and high (4.55 N/m(2) = 45.5 dynes/cm(2)) shear stresses. No significant differences in VWF morphology were found, the molecules were oriented almost randomly, and there were no clear signs of VWF "uncoiling" either at a high or at a low shear regime. After perfusing 1 microg/mL VWF for 5 minutes, surface coverage at high shear was almost twice the one seen at low shear, and some larger and more irregularly shaped VWF molecules could be seen at high shear. This difference disappeared, however, at 15 minutes of perfusion and was probably caused by diffusion kinetics. Moreover, the presence of 68 x 10(9)/L washed fixed platelets in the perfusate did not have any visible effect on VWF morphology at high versus low shear stress. These findings suggest that shear stress does not influence significantly the overall molecular morphology of VWF during its binding to collagen-coated surface and are consistent with a constitutively expressed affinity of collagen-bound VWF for glycoprotein Ib. PMID- 11877280 TI - The tumor vascular targeting agent combretastatin A-4-phosphate induces reorganization of the actin cytoskeleton and early membrane blebbing in human endothelial cells. AB - Combretastatin A-4-phosphate (CA-4-P) is a tubulin-binding compound currently in clinical trial as a tumor vascular-targeting agent. In endothelial cells, CA-4-P is known to cause microtubule depolymerization, but little is known about its subsequent effects on cell morphology and function. Here, we demonstrate that within minutes of endothelial cell exposure to CA-4-P, myosin light chain (MLC) was phosphorylated, leading to actinomyosin contractility, assembly of actin stress fibers, and formation of focal adhesions. These cytoskeletal alterations appeared to be a consequence of Rho activation, as they were abolished by either the Rho inhibitor C3 exoenzyme or Rho-kinase inhibitor Y-27632. In response to CA 4-P, some cells rapidly assumed a blebbing morphology in which F-actin accumulated around surface blebs, stress fibers misassembled into a spherical network surrounding the cytoplasm, and focal adhesions appeared malformed. Blebbing was associated with decreased cell viability and could be inhibited by Rho/Rho-kinase inhibitors or by blocking the CA-4-P-mediated activation of stress activated protein kinase-2/p38. The extracellular-regulated kinases 1 and 2 (ERK 1/2) were shown to protect against blebbing since blebbing was attenuated on ERK 1/2 stimulation and was up-regulated by specific inhibition of ERK-1/2 activation. The use of MLC kinase (MLCK) and myosin adenosine triphosphatase inhibitors led us to propose a role for MLCK and myosin activity independent of MLC phosphorylation in regulating the blebbing process. CA-4-P-mediated contractility and blebbing were associated with a Rho-dependent increase in monolayer permeability to dextrans, suggesting that such functional changes may be important in the rapid response of the tumor endothelium to CA-4-P in vivo. PMID- 11877282 TI - Identification of a tightly regulated hypoxia-response element in the promoter of human plasminogen activator inhibitor-1. AB - Plasminogen activator inhibitor-1 (PAI-1) plays a key role in control of coagulation and tissue remodeling and has been shown to be regulated by a number of cell stimuli, among those hypoxia. In this study we characterize the hypoxia mediated induction of PAI-1 in human hepatoma cell line HepG2. We found that PAI 1 is tightly regulated in a narrow oxygen gradient. After incubation at oxygen concentrations of 1% to 2%, a 60-fold increase in PAI-1 messenger RNA levels was observed, whereas mild hypoxic conditions of more than 3.5% did not appear to induce transcription. Moreover, increased levels of PAI-1 protein were observed after incubation at low oxygen tensions. Through sequence analysis, several putative hypoxia-response elements (HREs 1-5) were identified in the human PAI-I promoter. Reporter gene assays showed that the HRE-2 (-194 to -187) was necessary and sufficient for the hypoxia-mediated response. By electrophoretic mobility assay we observed hypoxia-dependent binding of a protein complex to the HRE-2 motif. Further analysis demonstrated that HRE-2 was specifically recognized by the hypoxia-inducible transcription factor 1alpha-arylhydrocarbon nuclear translocator complex. Taken together, our data demonstrate that hypoxia-induced transcription is mediated through HIF-1 interaction with the HRE-2 site of the human PAI-1 promoter. PMID- 11877283 TI - Anatomic location and T-cell stimulatory functions of mouse dendritic cell subsets defined by CD4 and CD8 expression. AB - Mouse spleen contains CD4+, CD8alpha+, and CD4-/CD8alpha- dendritic cells (DCs) in a 2:1:1 ratio. An analysis of 70 surface and cytoplasmic antigens revealed several differences in antigen expression between the 3 subsets. Notably, the Birbeck granule-associated Langerin antigen, as well as CD103 (the mouse homologue of the rat DC marker OX62), were specifically expressed by the CD8alpha+ DC subset. All DC types were apparent in the T-cell areas as well as in the splenic marginal zones and showed similar migratory capacity in collagen lattices. The 3 DC subtypes stimulated allogeneic CD4+ T cells comparably. However, CD8alpha+ DCs were very weak stimulators of resting or activated allogeneic CD8+ T cells, even at high stimulator-to-responder ratios, although this defect could be overcome under optimal DC/T cell ratios and peptide concentrations using CD8+ F5 T-cell receptor (TCR)-transgenic T cells. CD8alpha- or CD8alpha+ DCs presented alloantigens with the same efficiency for lysis by cytotoxic T lymphocytes (CTLs), and their turnover rate of class I-peptide complexes was similar, thus neither an inability to present, nor rapid loss of antigenic complexes from CD8alpha DCs was responsible for the low allostimulatory capacity of CD8alpha+ DCs in vitro. Surprisingly, both CD8alpha+ DCs and CD4-/CD8 DCs efficiently primed minor histocompatibility (H-Y male antigen) cytotoxicity following intravenous injection, whereas CD4+ DCs were weak inducers of CTLs. Thus, the inability of CD8alpha+ DCs to stimulate CD8+ T cells is limited to certain in vitro assays that must lack certain enhancing signals present during in vivo interaction between CD8alpha+ DCs and CD8+ T cells. PMID- 11877284 TI - Partial impairment of interleukin-12 (IL-12) and IL-18 signaling in Tyk2 deficient mice. AB - Tyk2 is activated in response to interleukin-12 (IL-12) and is essential for IL 12-induced T-cell function, including interferon-gamma (IFN-gamma) production and Th1 cell differentiation. Because IL-12 is a stimulatory factor for natural killer (NK) cell-mediated cytotoxicity, we examined whether tyk2 is required for IL-12-induced NK cell activity. IL-12-induced NK cell activity in cells from tyk2 deficient mice was drastically reduced compared to that in cells from wild-type mice. IL-18 shares its biologic functions with IL-12. However, the molecular mechanism of IL-18 signaling, which activates an IL-1 receptor-associated kinase and nuclear translocation of nuclear factor-kappaB, is different from that of IL 12. We next examined whether biologic functions induced by IL-18 are affected by the absence of tyk2. NK cell activity and IFN-gamma production induced by IL-18 were reduced by the absence of tyk2. Moreover, the synergistic effect of IL-12 and IL-18 for the production of IFN-gamma was also abrogated by the absence of tyk2. This was partially due to the absence of any up-regulation of the IL-18 receptor treated with IL-12, and it might suggest the presence of the cross-talk between Jak-Stat and mitogen-activated protein kinase pathways in cytokine signaling. PMID- 11877285 TI - ALK as a novel lymphoma-associated tumor antigen: identification of 2 HLA-A2.1 restricted CD8+ T-cell epitopes. AB - Oncogenic anaplastic lymphoma kinase (ALK) fusion proteins (NPM/ALK and associated variants) are expressed in about 60% of anaplastic large cell lymphomas (ALCLs) but are absent in normal tissues. In this study, we investigated whether ALK, which is expressed at high levels in lymphoma cells, could be a target for antigen-specific cell-mediated immunotherapy. A panel of ALK-derived peptides was tested for their binding affinity to HLA-A*0201 molecules. Binding peptides were assessed for their capacity to elicit a specific immune response mediated by cytotoxic T lymphocytes (CTLs) both in vivo, in HLA A*0201 transgenic mice, and in vitro in the peripheral blood lymphocytes (PBLs) from healthy donors. Two HLA-A*0201-restricted CTL epitopes, p280-89 (SLAMLDLLHV) and p375-86 (GVLLWEIFSL), both located in the ALK kinase domain were identified. The p280-89- and p375-86-induced peptide-specific CTL lines were able to specifically release interferon-gamma (IFN-gamma) on stimulation with ALK peptide pulsed autologous Epstein-Barr virus-transformed B cells (LCLs) or T2 cells. Anti ALK CTLs lysed HLA-matched ALCL and neuroblastoma cell lines endogenously expressing ALK proteins. CTL activity was inhibited by anti-HLA-A2 monoclonal antibody CR11.351, consistent with a class I-restricted mechanism of cytotoxicity. These results show the existence of functional anti-ALK CTL precursors within the peripheral T-cell repertoire of healthy donors, clearly indicating ALK as a tumor antigen and ALK-derived peptides, p280-89 and p375-86, as suitable epitopes for the development of vaccination strategies. PMID- 11877286 TI - Leukemic target susceptibility to natural killer cytotoxicity: relationship with BCR-ABL expression. AB - Chronic myeloid leukemia is a clonal myeloproliferative expansion of transformed primitive hematopoietic progenitor cells characterized by high-level expression of BCR-ABL chimeric gene, which induces growth factor independence. However, the influence of BCR-ABL expression on cell-mediated cytotoxicity is poorly understood. In the present study, we asked whether BCR-ABL expression interferes with leukemic target sensitivity to natural killer (NK) cell cytolysis. Our approach was based on the use of 2 BCR-ABL transfectants of the pluripotent hematopoietic cell line UT-7 expressing low (UT-7/E8, UT-7/G6) and high (UT-7/9) levels of BCR-ABL. As effector cells, we used CD56(bright), CD16-, CD2- NK cells differentiated in vitro from CD34 cord blood progenitors. We demonstrated that BCR-ABL transfectants UT-7/9 were lysed by NK cells with a higher efficiency than parental and low UT-7/E8.1 and UT-7/G6 transfectants. This enhanced susceptibility to lysis correlated with an increase in expression of intercellular adhesion molecule 1 (ICAM-1) by target cells. Treatment of UT-7/9 cells by STI571 (a specific inhibitor of the abl kinase) resulted in a decrease in NK susceptibility to lysis and ICAM-1 down-regulation in target cells. Furthermore, the constitutive activation of nuclear factor-kappaB (NF-kappaB) detected in BCR-ABL transfectant UT-7/9, was significantly attenuated when cells were treated by STI571. Interestingly, inhibition of NF-kappaB activation by BAY11-67082 (a specific NF-kappaB inhibitor) resulted in down-regulation of ICAM 1 expression and a subsequent decrease in NK-induced killing of UT-7/9 transfectants. Our results show that oncogenic transformation by BCR-ABL may increase susceptibility of leukemic progenitors to NK cell cytotoxicity by a mechanism involving overexpression of ICAM-1 as a consequence of NF-kappaB activation. PMID- 11877287 TI - Interleukin-17 inhibits tumor cell growth by means of a T-cell-dependent mechanism. AB - Interleukin 17 (IL-17) is a proinflammatory cytokine produced by activated CD4(+) memory T cells. We previously showed that IL-17 increased the growth rate of human cervical tumors transplanted into athymic nude mice. To address the possible role of T cells in the biologic activity of IL-17 for tumor control, we grafted 2 murine hematopoietic immunogenic tumors (P815 and J558L) transfected with a complementary DNA encoding murine IL-17 into syngeneic immunocompetent mice. We found that growth of the 2 IL-17-producing tumors was significantly inhibited compared with that of mock-transfected tumors. In contrast to the antitumor activity of IL-17 observed in immunocompetent mice, we observed no difference in the in vivo growth of IL-17-transfected or mock-transfected P815 cells (P815-IL-17 and P815-Neo, respectively) transplanted into nude mice. We then showed that IL-17 increased generation of specific cytolytic T lymphocytes (CTLs) directed against the immunodominant antigens from P815 called A, B, C, D, and E, since all mice injected with P815-IL-17 developed a P815-specific CTL response, whereas only 6 of 16 mice immunized with P815-Neo had a specific CTL response against the antigens. The induction of CTLs was associated with establishment of a tumor-protective immunity. These experiments suggest that T lymphocytes are involved in the antitumor activity of IL-17. Therefore, IL-17, like other cytokines, appears to be a pleiotropic cytokine with possible protumor or antitumor effects on tumor development, which often depends on the immunogenicity of tumor models. PMID- 11877288 TI - Effects of administration of progenipoietin 1, Flt-3 ligand, granulocyte colony stimulating factor, and pegylated granulocyte-macrophage colony-stimulating factor on dendritic cell subsets in mice. AB - We studied the effects of administration of several cytokines, including progenipoietin-1 (ProGP-1), Flt-3 ligand (FL), granulocyte colony-stimulating factor (G-CSF), and granulocyte-macrophage colony-stimulating factor in a pegylated form (pGM-CSF), on dendritic cell (DC) populations in mouse spleen. ProGP-1 produced the most striking increase in overall DC numbers, apparently more than its constituent FL and G-CSF components. However, the expansion in DC numbers was strongly subpopulation selective, with ProGP-1 and FL producing selective expansion of CD8+ DCs, whereas pGM-CSF produced selective expansion of CD8- DCs. Surprising differences were observed between the effects of murine and human recombinant FL preparations on murine DCs. Many of the biologic functions of the DC subpopulations expanded by cytokines remained intact, including the capacity of the ProGP-1- and FL-expanded CD8+ DCs to produce the T-helper-1 biasing cytokine interleukin 12 (IL-12). However, the expanded DCs from all but G CSF-treated mice were deficient in the ability to make interferon gamma, and the CD8+ DCs produced with pGM-CSF treatment had an abrogated capacity to form bioactive IL-12. Such selective expansion of DC populations and alterations in their cytokine-secretion capacity have implications for clinical use of the studied cytokines in immune modulation. PMID- 11877289 TI - Long-lasting memory-resting and memory-effector CD4+ T cells in human X-linked agammaglobulinemia. AB - Conflicting results obtained from animal studies suggest that B cells play a role in maintaining long-term T-cell memory and in skewing T-cell response toward a T helper 2 (T(H)2) phenotype. X-linked agammaglobulinemia (XLA) is a genetic human disease characterized by the lack of circulating B cells due to the mutation of Bruton tyrosine kinase. This disease thus represents a unique model for studying the role of B lymphocytes in regulating T-cell functions in humans. To this aim, we analyzed hepatitis B envelope antigen (HBenvAg)-specific T-cell memory in a series of XLA patients vaccinated against hepatitis B virus (HBV). We found HBenvAg-specific T lymphocytes producing interferon-gamma, interleukin-4, or both in the peripheral blood of XLA patients up to at least 24 months after completing the standard anti-HBV immunization protocol. The HBenvAg-specific T-cell frequencies and the percentage of patients with these responses were not significantly different from healthy vaccinated controls. By combining cell purification and enzyme-linked immunospot assay, we found that effector CD27- T cells, which promptly produced cytokines in response to antigen (Ag), and memory resting CD27+ T cells, which required Ag restimulation to perform their functions, were maintained in both XLA patients and controls for up to 24 months after the last vaccination boost. These data strongly suggest that B cells are not an absolute requirement for the generation of effective T-cell memory in humans, nor do they seem to influence T(H)1/T(H)2 balance. PMID- 11877290 TI - A novel costimulatory signaling in human T lymphocytes by a splice variant of CD28. AB - We have characterized a splice variant (isoform) of the human CD28 T cell costimulatory receptor. The nucleotide sequence of this CD28 isoform was identical to that of CD28 in the signal peptide, the transmembrane domain, and the cytoplasmic tail, but it was missing a large segment of the extracellular ligand-binding domain, which is encoded by the second exon. This isoform (CD28i), whose message level exceeded 25% of CD28, was a transmembrane homodimer. CD28i was found noncovalently associated with CD28 and was tyrosine-phosphorylated/PI3 kinase-complexed following the crosslinking of CD28, and the CD28 costimulatory signal was enhanced in T cells expressing CD28i. These data demonstrate that CD28i, via noncovalent association with CD28, plays a role as a costimulatory signal amplifier in human T cells. PMID- 11877291 TI - Negative effect of CTLA-4 on induction of T-cell immunity in vivo to B7-1+, but not B7-2+, murine myelogenous leukemia. AB - B7 molecules provide important costimulatory signals to T cells, and B7 genes have been introduced into B7-negative tumor cells to enhance their immunogenicity. However, the role of B7 molecules in inducing tumor immunity is controversial because of conflicting results and reports of differential signaling through the B7 molecules and their ligands CD28 and CTLA-4. In this study, we compared the effect of B7-1 (CD80) and B7-2 (CD86) on the induction of T-cell immunity to C1498, a murine myelogenous leukemia. When cultured with exogenous cytokines in vitro, C1498/B7-1 and C1498/B7-2 induced syngeneic CD8+ T cells to kill parental C1498. In vivo, C1498/B7-1 grew progressively after subcutaneous injection, whereas C1498/B7-2 completely regressed after transient growth in naive mice. Spontaneous rejection of C1498/B7-2 resulted in immunity to challenge doses of C1498 and C1498/B7-1. Antibody-depletion studies in vivo showed that CD8+ T cells rejected C1498/B7-2, whereas only natural killer cells affected the growth of C1498/B7-1. Two approaches were used to determine whether preferential interaction of B7-1 with CTLA-4 contributed to the failure of C1498/B7-1 to activate CD8+ T cells in vivo. First, CTLA-4 specific monoclonal antibody was used to block B7-1-CTLA-4 interaction. Second, CTLA-4 deletional mutant (-/-) bone marrow chimeras were used as tumor hosts. In both systems, there was a significant increase in the rate of rejection of C1498/B7-1 tumors. Resistance to C1498/B7-1 in CTLA-4(minus sign/minus sign) hosts was mediated by CD8+ T cells. Blocking or deletion of CTLA-4 did not affect the growth of parental C1498, indicating that B7-1 was important for the induction of CD8+ T cell immunity in the absence of CTLA-4. PMID- 11877292 TI - The heterogeneity shown by human plasma cells from tonsil, blood, and bone marrow reveals graded stages of increasing maturity, but local profiles of adhesion molecule expression. AB - Plasma cells (PCs) are the final B-cell differentiation stage. Recent evidence reveals relevant functional differences within the PC compartment. In rodents, early PCs formed in secondary lymphoid tissues show enhanced apoptosis and short life span, whereas PCs present in a final destination organ, such as the bone marrow (BM), have reached a stable prolonged survival state. BM PCs arrive at this organ as a circulating precursor whose cellular nature remains uncertain. An initial aim of this study was to characterize this circulating cell. We hypothesized that antibody-secreting cells detectable in the human blood after immunization might be a candidate precursor. These cells were obtained from the blood of volunteers immunized 6 days earlier with tetanus toxoid (tet), and they were unambiguously identified as PCs, as demonstrated by their expression of the CD38(h) phenotype, by morphology, by immunoglobulin (Ig) intracytoplasmic staining, and by IgG-tet-secreting capacity in vitro. In addition, by using the common CD38(h) feature, human PCs from tonsil (as a possible source of early PCs), from blood from tet-immunized donors (as the putative precursors of BM PCs), and from BM (as a deposit organ) have been purified and their phenotypes compared. The results show that a variety of differentiation molecules, proteins involved in the control of apoptosis, the B-cell transcription factors, positive regulatory domain I-binding factor 1/B lymphocyte-induced maturation protein 1 and B cell-specific activating protein and, at least partially, the chemokine receptor CXCR4 were expressed by human PCs following a gradient of increasing maturity in the direction: tonsil-->blood-->BM. However, PCs from these different organs showed a local pattern of adhesion molecule expression. These observations are discussed in light of the complex physiology of the human PC compartment. PMID- 11877293 TI - Intracellular regulation of tumor necrosis factor-related apoptosis-inducing ligand-induced apoptosis in human multiple myeloma cells. AB - Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL, Apo2 ligand) effectively kills multiple myeloma (MM) cells in vitro irrespective of refractoriness to dexamethasone and chemotherapy. Because clinical trials with this anticancer agent are expected shortly, we investigated the signaling pathway of TRAIL-induced apoptosis in MM. We detected rapid cleavage of caspases-8, -9, 3, and -6, as well as the caspase substrates poly(ADP-ribose) polymerase (PARP) and DNA fragmentation factor-45 (DFF45), but not caspase-10, upon TRAIL treatment in sensitive MM cells, pointing to caspase-8 as the apical caspase of TRAIL signaling in MM cells. These phenomena were not observed or were significantly delayed in TRAIL-resistant MM cells, suggesting that resistance may arise from inhibition at the level of caspase-8 activation. Higher levels of expression for various apoptosis inhibitors, including FLICE-inhibitory protein (FLIP), and lower procaspase-8 levels were present in TRAIL-resistant cells and sensitivity was restored by the protein synthesis inhibitor cycloheximide (CHX) and the protein kinase C (PKC) inhibitor bisindolylmaleimide (BIM), which both lowered FLIP and cellular inhibitor of apoptosis protein-2 (cIAP-2) protein levels. Forced expression of procaspase-8 or FLIP antisense oligonucleotides also sensitized TRAIL-resistant cells to TRAIL. Moreover, the cell permeable nuclear factor (NF)-kappaB inhibitor SN50, which sensitizes TRAIL-resistant cells to TRAIL, also inhibited cIAP2 protein expression. Finally, CHX, BIM, and SN50 facilitated the cleavage and activation of procaspase-8 in TRAIL-resistant cells, confirming that inhibition of TRAIL-induced apoptosis occurs at this level and that these agents sensitize MM cells by relieving this block. Our data set a framework for the clinical use of approaches that sensitize MM cells to TRAIL by agents that inhibit FLIP and cIAP-2 expression or augment caspase-8 activity. PMID- 11877294 TI - Requirements of src family kinase activity associated with CD45 for myeloma cell proliferation by interleukin-6. AB - Specific intracellular signals mediated by interleukin-6 (IL-6) receptor complexes, such as signal transducer and activator of transcription 3 (STAT 3) and extracellular signal-regulated kinase (ERK) 1/2, are considered to be responsible for inducing a variety of cellular responses. In multiple myeloma, IL 6 only enhanced the proliferation of CD45+ tumor cells that harbored the IL-6 independent activation of src family kinases even though STAT3 and ERK1/2 could be activated in response to IL-6 in both CD45+ and CD45(minus sign) cells. Furthermore, the IL-6-induced proliferation of CD45+ U266 myeloma cells was significantly suppressed by Lyn-specific antisense oligodeoxynucleotides or a selective src kinase inhibitor. These results indicate that the activation of both STAT3 and ERK1/2 is not enough for IL-6-induced proliferation of myeloma cell lines that require src family kinase activation independent of IL-6 stimulation. Thus, the activation of the src family kinases associated with CD45 expression is a prerequisite for the proliferation of myeloma cell lines by IL-6. We propose a mechanism for IL-6-induced cell proliferation that is strictly dependent upon the cellular context in myelomas. PMID- 11877295 TI - Vascular endothelial growth factor (VEGF)-C signaling through FLT-4 (VEGFR-3) mediates leukemic cell proliferation, survival, and resistance to chemotherapy. AB - Similar to solid tumors, growth of leukemias may also be angiogenesis dependent. Furthermore, tyrosine kinase receptors specific to endothelial cells are expressed on certain subsets of leukemias. We have previously demonstrated the existence of a VEGF/VEGFR-2 autocrine loop on leukemic cells that supports their growth and migration. Here, we demonstrate that in response to leukemia-derived proangiogenic and proinflammatory cytokines such as basic fibroblast growth factor and IL-1, endothelial cells release increasing amounts of another vascular endothelial growth factor (VEGF) family member, VEGF-C. In turn, interaction of VEGF-C with its receptor VEGFR-3 (FLT-4) promotes leukemia survival and proliferation. We demonstrate in 2 cell lines and 5 FLT-4(+) leukemias that VEGF C and a mutant form of the molecule that lacks the KDR-binding motif induce receptor phosphorylation, leukemia proliferation, and increased survival, as determined by increased Bcl-2/Bax ratios. Moreover, VEGF-C protected leukemic cells from the apoptotic effects of 3 chemotherapeutic agents. Because most leukemic cells release proangiogenic as well as proinflammatory cytokines, our data suggest that the generation of a novel paracrine angiogenic loop involving VEGF-C and FLT-4 may promote the survival of a subset of leukemias and protect them from chemotherapy-induced apoptosis. These results identify the VEGF-C/FLT-4 pathway as a novel therapeutic target for the treatment of subsets of acute leukemia. PMID- 11877297 TI - Tumor cell dissemination in follicular lymphoma. AB - The derivation of follicular lymphomas (FLs) from germinal centers is not only supported by their morphologic appearance with a nodular growth pattern and a germinal center-like cellular composition, but also by the presence of ongoing somatic hypermutation (a germinal center B cell-specific process) during their clonal expansion. The intraclonal sequence diversity of the tumor cells and their follicular growth pattern allows one to analyze lymphoma cell dissemination and the way the tumor "metastasizes" to distinct follicles. In the present study, we analyzed individual follicles of 3 FLs by micromanipulation of single cells from individual lymphoma follicles and amplification of immunoglobulin V region genes. Genealogical trees for the V(H) and the V(L) gene rearrangements were constructed to analyze the clonal relationship among individual cells of 3 distinct follicles of each case. In all 3 cases there is evidence that distinct tumor follicles are founded by many tumor cells, suggesting that there is extensive migration of the tumor cells among follicles. The observation that the tumor cells of FLs retain their follicular growth patterns despite this cellular migration supports the idea that they depend on the follicular microenvironment for their clonal expansion. PMID- 11877296 TI - Oncogenesis of multiple myeloma: 14q32 and 13q chromosomal abnormalities are not randomly distributed, but correlate with natural history, immunological features, and clinical presentation. AB - Multiple myeloma (MM) is a plasma-cell malignancy characterized by marked epidemiological, biological, and clinical heterogeneity. The goal of this study was to find a genetic basis for this heterogeneity. Using fluorescence in situ hybridization, we analyzed a prospective cohort of 901 patients with various plasma-cell disorders--monoclonal gammopathies of undetermined significance, smoldering MM, MM, and primary plasma-cell leukemia--for genetic abnormalities involving the 13q14 and 14q32 chromosomal regions; the patients were consecutively enrolled in the Intergroupe Francophone du Myelome clinical trials, We performed statistical analyses comparing these chromosomal abnormalities in terms of immunological (ie, immunoglobulin types and light-chain subtypes) and clinical status and, to some extent, prognostic features. It was found that 14q32 translocations and del(13) are the most frequent chromosomal abnormalities, observed in 75% and 45% of the patients, respectively, and are not randomly distributed, but interconnected. Second, correlations between them allowed us to define 4 major genetic categories of patients: (1) patients lacking any 14q32 abnormality (25%) and generally also lacking del(13); (2) patients presenting either t(4;14) or t(14;16), almost always associated with a del(13) (15% of patients); (3) patients with other 14q32 abnormalities and presenting del(13) (25%); and (4) patients with other 14q32 abnormalities but not presenting del(13) (35%). Third, we show that this genetic stratification is highly correlated with immunological status and clinical presentation and with some major prognostic factors. For the first time, this study gives genetic support to the heterogeneity observed in patients with MM and demonstrates that the 14q32 and 13q chromosomal abnormalities are not randomly distributed. The strong correlations we found might be the basis for a novel genetic classification of MM, as has been previously demonstrated for leukemias and lymphomas. Furthermore, our study supports different models for MM oncogenesis. PMID- 11877298 TI - Retinoic acid-induced cell cycle arrest of human myeloid cell lines is associated with sequential down-regulation of c-Myc and cyclin E and posttranscriptional up regulation of p27(Kip1). AB - All-trans retinoic acid (ATRA) is a potential therapeutic agent for the treatment of hematopoietic malignancies, because of its function as an inducer of terminal differentiation of leukemic blasts. Although the efficacy of ATRA as an anticancer drug has been demonstrated by the successful treatment of acute promyelocytic leukemia (APL), the molecular mechanisms of ATRA-induced cell cycle arrest of myeloid cells have not been fully investigated. In this study, we show that the onset of ATRA-induced G(0)/G(1) arrest of human monoblastic U-937 cells is linked to a sharp down-regulation of c-Myc and cyclin E levels and an increase in p21(WAF1/CIP1) expression. This is followed by an increase in p27(Kip1) protein expression due to enhanced protein stability. The importance of an early decrease in Myc expression for these events was demonstrated by the failure of a U-937 subline with constitutive exogenous expression of v-Myc to cell cycle arrest and regulate cyclin E and p27(Kip1) in response to ATRA. Preceding the initiation of G(1) arrest, a transient rise in retinoblastoma protein (pRb), p107, and cyclin A levels was detected. Later, a rapid fall in the levels of cyclins A and B and a coordinate dephosphorylation of pRb at Ser780, Ser795, and Ser807/811 coincided with the accumulation of cells in G(1). These results thus identify a decrease in c-Myc and cyclin E levels and a posttranscriptional up regulation of p27(Kip1) as important early changes, and position them in the complex chain of events regulating ATRA-induced cell cycle arrest of myeloid cells. PMID- 11877299 TI - Mechanical properties of rat bone marrow and circulating neutrophils and their responses to inflammatory mediators. AB - Neutrophils are continuously released from the bone marrow (BM), and this release is accelerated during inflammation. This study compared the mechanical properties of mature neutrophils within the BM and the circulating blood, as well as the role of microtubule rearrangement in the release of neutrophils from the BM in rats. Neutrophils isolated from the BM were stiffer than neutrophils in the circulating blood, using magnetic twisting cytometry. BM neutrophils also contained more F-actin within the submembrane region than circulating neutrophils when examined using confocal microscopy, suggesting that mature quiescent neutrophils within the BM are stiffer than circulating neutrophils because of increased formation of F-actin beneath the plasma membrane. Complement protein 5 fragments or formylmethionyl-leucylphenylalanine (fMLP) induced a stiffening response within 2 minutes that was greater in circulating than in BM neutrophils. This stiffening required F-actin formation within the submembrane region but not microtubule rearrangement in both circulating and BM neutrophils. fMLP-induced shape changes were more pronounced in circulating than in BM neutrophils, which showed fewer and smaller pseudopods and fewer membrane irregularities. In vivo, fMLP induced neutropenia, sequestration of neutrophils within the pulmonary capillaries, and release of neutrophils from the BM. Studies using colchicine demonstrated that rearrangement of microtubules was not required for any of these processes but was required for normal trafficking of neutrophils through the pulmonary capillaries. PMID- 11877300 TI - Human eosinophils produce neurotrophins and secrete nerve growth factor on immunologic stimuli. AB - Neurotrophins, such as nerve growth factor (NGF) and neurotrophin-3 (NT-3), are essential for development, function, and survival of peripheral sympathetic and sensory neurons. Most eosinophilic leukocytes in the human body are localized in mucosal tissues; however, the roles of eosinophils in human diseases are not fully understood. We found that human eosinophils constitutively express messenger RNA for NGF and NT-3, synthesize and store these proteins intracellularly, and continuously replenish them. Incubation of eosinophils with a transcription inhibitor, actinomycin D, for 8 hours completely depletes intracellular NGF and NT-3. New synthesis of NGF is enhanced by Fc-receptor mediated stimuli, such as immunoglobulin (Ig)A and IgG immune complexes; in contrast, production of NT-3 is not affected by these stimuli. Notably, supernatants of eosinophils stimulated with IgA immune complex and interleukin 5 promote neurite extension of the PC-12 pheochromocytoma cell line; this effect is abolished by pretreatment of the supernatants with anti-NGF-neutralizing antibody. By enzyme-linked immunosorbent assay, substantial amounts of NGF protein are also detected in the supernatants of stimulated eosinophils. Furthermore, in patients with seasonal allergic rhinitis, the concentrations of NGF in nasal secretions correlate with the magnitudes of eosinophilic inflammation in the airway, suggesting a potential clinical implication of eosinophil NGF. Our observations propose a new pathologic mechanism by which eosinophils may contribute to enhanced neurologic responses in patients with allergic diseases and other eosinophilic disorders. Alternatively, eosinophils may play important roles in maintenance and restoration of homeostatic functions of mucosal tissues through the pleitropic activities of NGF. PMID- 11877301 TI - High-affinity binding sites for heparin generated on leukocytes during apoptosis arise from nuclear structures segregated during cell death. AB - During cell death of human cultured leukocytes (Jurkat, HL-60, THP-1, U937) and freshly prepared leukocytes, we observed a greater than 100-fold increase in the affinity of apoptotic and necrotic cells for fluorescein isothiocyanate (FITC) heparin in comparison with live cells. Binding of FITC-heparin was reversed in the presence of high ionic strength, unlabeled heparan sulfate, and heparin and pentosan polysulfate, but not in the presence of chondroitin and dermatan sulfates. During the course of cell death, the increase in the percentage of cells positive for annexin V binding correlated with the increase in the population positive for binding FITC-heparin. Confocal microscopy demonstrated that heparin binding to dead cells was restricted to 1 or 2 small domains on the surfaces of apoptotic cells and to larger, but still discrete, areas that did not localize with chromatin on ruptured necrotic cells. The heparin-binding domains originated from the nucleus and may correspond to the ribonucleoprotein containing structures that have recently been shown to segregate within the nucleus of cells and to move onto the cell membrane. We observed that phagocytosis of dead Jurkat cells by monocyte-derived macrophages was blocked when the heparin-binding capacity of the dead cells was saturated by the addition of pentosan polysulfate. From this we concluded that the ability of dead cells to bind to heparan sulfate proteoglycans on the surfaces of macrophages may assist in phagocytic clearance. PMID- 11877302 TI - Differential effect of CD28 versus B7 blockade on direct pathway of allorecognition and self-restricted responses. AB - Immunosuppression with B7 antagonists might have 2 opposite effects: reducing T cell costimulation through CD28 but also preventing CTLA-4 from transmitting its negative regulatory signal. We therefore hypothesized that a selective blockade of CD28 might be qualitatively different from blocking B7. It was previously reported that CD28 modulation prolongs allograft survival in the rat and reverses induction of experimental autoimmune encephalomyelitis in mice. However, whether CD28 or B7 blockade results in similar immunosuppression on alloimmune and self restricted responses to soluble antigens has not yet been investigated. Here, we addressed this issue in vitro with antagonist anti-CD28 Fab fragments and in vivo using the modulating anti-rat JJ319 monoclonal antibody. As in the inhibition of B7 with CTLA4 immunoglobulin, anti-CD28 Fab fragments inhibited allogenic T-cell proliferation in mixed cultures. In vivo modulation of CD28 blocked the expansion of alloreactive T cells and promoted their apoptosis. In contrast, selective blockade of CD28 did not modify T-cell proliferative responses and antibody production to soluble antigens, whereas blocking B7 with CTLA4 immunoglobulin did. Our data show that blocking CD28, while leaving CTLA4-B7 interactions undisturbed, inhibits alloreactive CD4+ T-cell expansion but does not modify the response to nominal antigens presented in the context of a self-major histocompatibility complex. That B7 engagement is needed for self-restricted responses whereas engagement of CD28 is not essential adds to the suggestion that another unidentified ligand of B7 might deliver a costimulatory signal in the absence of CD28. PMID- 11877303 TI - Addition of a second, different allogeneic graft accelerates white cell and platelet engraftment after T-cell-depleted bone marrow transplantation. AB - Significant delays in engraftment and lymphoid recovery are the 2 major challenges in cord blood transplantation. The cause for this delay is presumed to be the low numbers of hematopoietic precursors found in one unit of cord blood. One approach to increase the stem cell doses could be to combine cord blood units from different donors differing at the major histocompatibility complex (MHC). As a first step toward this goal, the kinetics of hematologic engraftment and immune reconstitution were compared between 1 unit (2.5 x 10(6) cells) of T-cell depleted bone marrow cells from a single donor (C57BL/6 [H2(b)] or SJL/J [H2(s)]) and 2 units from different donors (C57BL/6 + SJL/J) after transplantation into lethally irradiated (8.5 Gy) BALB/c recipients (H2(d)). Addition of T-cell depleted bone marrow from an MHC-mismatched allogeneic donor doubled the white blood counts compared with recipients of one single unit on days +10 and +14. Similar effects were also observed on platelets. The beneficial effect of additional cells on peripheral T-cell counts were first observed on day +14. Cells both from donors (C57BL/6 and/or SJL/J) and recipients (BALB/c) contributed to myeloid and lymphoid reconstitution. The chimeras containing cells from 3 strains of mice were able to mount a recall immune response. Our data suggest that combining stem cells from MHC-mismatched allogeneic donors is feasible, that it has beneficial effects on myeloid engraftment and T-cell phenotypic recovery, and that the long-term stable mixed chimeras are immunologically normal following T-cell-depleted bone marrow transplantation. PMID- 11877304 TI - Combined treatment with temozolomide and poly(ADP-ribose) polymerase inhibitor enhances survival of mice bearing hematologic malignancy at the central nervous system site. AB - Temozolomide (TZM) is a DNA-methylating agent that has recently been introduced into various clinical trials for treatment of solid or hematologic neoplasias, including brain lymphomas. In the current study, we have investigated whether the antitumor activity of TZM could be selectively enhanced at the central nervous system (CNS) site by intracerebral injection of a poly(ADP-ribose) polymerase (PARP) inhibitor. Mice were injected intracranially with lymphoma cells. The PARP inhibitor NU1025 (1 mg/animal) was delivered intracerebrally, whereas TZM was given as a single or a fractionated dose of 200 mg/kg by intraperitoneal administration. Results indicated that this drug combination significantly enhanced the survival of tumor-bearing mice and that this fractionated modality of treatment was the most effective schedule. Increased survival time was related to a marked reduction of tumor growth, as evidenced by histologic studies. Treatment with TZM alone was ineffective. This is the first report exploring in vivo the combination of TZM with PARP inhibitor for intracerebral neoplasias. PMID- 11877305 TI - Campath-1H and fludarabine in combination are highly active in refractory chronic lymphocytic leukemia. AB - Campath-1H (alemtuzumab) is the most effective monoclonal antibody in single agent use in B-cell chronic lymphocytic leukemia (CLL) with reported response rates of 33% to 70%. Combination therapy is now the conventional treatment for most hematologic malignancies. Monoclonal antibody treatments may sensitize tumor cells to subsequent chemotherapy. We report the combination of Campath-1H with fludarabine in patients with CLL refractory to each agent used singly. Six patients who had received a median of 8 courses of fludarabine (range, 4-10 courses) and 16 weeks of Campath-1H (range, 8-32 weeks) were treated. Five patients responded, including one who had a complete response by National Cancer Institute criteria. The responses observed were better in each patient than responses after each agent used singly. Complete morphologic bone marrow responses were seen in 3 patients, including eradication of disease measured by sensitive flow cytometry in 2. Campath-1H combined with fludarabine is a highly promising novel therapy for refractory CLL. PMID- 11877306 TI - Histone deacetylase inhibitor FR901228 enhances adenovirus infection of hematopoietic cells. AB - Adenovirus infection of hematopoietic cells frequently requires high virus concentrations and long incubation times to obtain moderate infection levels because these cells have low levels of Coxsackie and adenovirus receptor (CAR) and alpha(v) integrin. The effect of treatment with FR901228 (depsipeptide), a histone deacetylase inhibitor in phase 2 clinical trials, was studied in K562 cells, granulocyte-colony-stimulating factor-mobilized peripheral blood mononuclear cells (PBMCs), and CD34+ peripheral blood stem cells (PBSCs). FR901228 increased CAR and alpha(v) integrin RNA levels and histone H3 acetylation. FR901228 treatment before adenovirus infection was associated with at least a 10-fold increase in transgene expression from a beta-galactosidase expressing adenoviral vector. More than 80% of the PBMCs or CD34+ PBSCs from 7 different donors were beta-galactosidase-positive after adenovirus infection with a multiplicity of infection of 10 for 60 minutes. Increased CAR, alpha(v) integrin, and acetylated histone H3 levels were observed in PBMCs from a patient treated with FR901228. These studies suggest that FR901228 can increase the efficiency of adenoviral infection in hematopoietic cells. PMID- 11877307 TI - Etanercept, a soluble tumor necrosis factor receptor, palliates constitutional symptoms in patients with myelofibrosis with myeloid metaplasia: results of a pilot study. AB - Patients with myelofibrosis with myeloid metaplasia (MMM) often experience debilitating constitutional symptoms such as drenching night sweats, profound fatigue, unexplained fevers, and unintentional weight loss. Tumor necrosis factor (TNF) contributes to organ fibrosis and hypercatabolic symptoms in a variety of disease states. We conducted an open-label pilot study of etanercept, a soluble TNF receptor, administered at a dose of 25 mg subcutaneously twice weekly for up to 24 weeks in 22 patients with MMM. Of 20 evaluable patients, 12 (60%) experienced an improvement in constitutional symptoms, and 4 (20%) had an objective response (improvement in peripheral cytopenias or spleen size). The degree of marrow fibrosis was unchanged, and only minor changes in overall marrow cellularity were observed. Toxicity was mild, with injection site reactions (20%) and minor infections (10%) as the most common side effects. One patient developed reversible pancytopenia. Etanercept may be useful for palliation of constitutional symptoms in MMM. PMID- 11877308 TI - Allogeneic blood cell transplantation following reduced-intensity conditioning is effective therapy for older patients with myelofibrosis with myeloid metaplasia. AB - Standard myeloablative conditioning prior to allogeneic hematopoietic stem cell (HSC) transplantation has been associated with significant toxicity in patients older than 45 years of age with myelofibrosis with myeloid metaplasia (MMM). We sought to evaluate the efficacy of a reduced-intensity conditioning regimen for allogeneic HSC transplantation in this setting. A regimen consisting of fludarabine (30 mg/m(2) intravenously daily for 5 days) and melphalan (70 mg/m(2) intravenously daily for 2 days) followed by transplantation of filgrastim mobilized peripheral blood cells from HLA-identical siblings was administered to 4 older patients (median age, 56 years; range, 48-58 years) with advanced MMM. All patients achieved prompt neutrophil and platelet engraftment and have experienced a significant regression of splenomegaly and bone marrow fibrosis. All now have normal bone marrow cellularity. With a median follow-up of 13 months (range, 11-19 months), all 4 patients are alive with stable full-donor hematopoietic chimerism. These results support the feasibility and effectiveness of reduced-intensity conditioning prior to allogeneic HSC transplantation for older patients with advanced MMM. PMID- 11877309 TI - Regression of lymphoproliferative disorder after treatment for hepatitis C virus infection in a patient with partial trisomy 3, Bcl-2 overexpression, and type II cryoglobulinemia. AB - A patient with type II cryoglobulinemic vasculitis and hepatitis C virus (HCV) infection presented with a leukemiclike proliferation of B cells bearing marginal zone B-cell phenotypic markers. A partial trisomy 3 (bands 3q11-29) and overexpression of Bcl-2 without t(14;18) translocation was detected in the monoclonal B cells that were classic rheumatoid factor-producing B cells bearing the WA cross-idiotype. Treatment with interferon-alpha produced a complete clinical remission and synchronous marked decreases in viremia and monoclonal B cell prevalence. This is the first report of partial trisomy 3 and Bcl-2 overexpression in type II cryoglobulinemic vasculitis associated with HCV infection. Further studies of HCV-infected patients with and without type II cryoglobulinemia are required to determine the prevalence and possible physiologic and/or pathophysiologic significance of these findings. PMID- 11877310 TI - Somatically mutated Ig V(H)3-21 genes characterize a new subset of chronic lymphocytic leukemia. AB - Recent studies on the immunoglobulin variable heavy chain (IgV(H)) genes have revealed that B-cell chronic lymphocytic leukemia (B-CLL) consists of at least 2 clinical entities with either somatically mutated or unmutated V(H) genes. We have analyzed the V(H) gene mutation status and V(H) gene usage in 119 B-CLL cases and correlated them to overall survival. A novel finding was the preferential use of the V(H)3-21 gene in mutated cases, whereas biased V(H)1-69 gene usage was found in unmutated cases as previously reported. Interestingly, the subset of mutated cases using the V(H)3-21 gene displayed distinctive genotypic/phenotypic characteristics with shorter average length of the complementarity determining region 3 and clonal expression of lambda light chains. In addition, this mutated subset showed significantly shorter survival than other mutated cases and a similar clinical course to unmutated cases. We therefore suggest that B-CLL cases with mutated V(H)3-21 genes may constitute an additional entity of B-CLL. PMID- 11877311 TI - Prognostic significance of cellular vascular endothelial growth factor expression in chronic phase chronic myeloid leukemia. AB - The impact of elevated vascular endothelial growth factor (VEGF) expression on the course of chronic myeloid leukemia (CML) is unknown. By radioimmunoassay, we measured pretreatment cellular VEGF protein in bone marrow samples from 184 (148 chronic and 36 accelerated/blastic phases) CML patients and found the levels to be 1.6-fold higher than in 31 normal control bone marrow samples (P =.000 01). No significant differences were found in VEGF levels by different phases of CML (P =.1). VEGF levels correlated with older age (P =.01) and higher platelet count (P =.0003), but also with smaller spleen size (P =.004), lower white blood cell count (P =.0006), and lower percentage of peripheral blasts (P =.04). With the use of Cox proportional hazard model and VEGF levels as a continuous variable, high VEGF levels correlated with shorter survival of patients in chronic CML (P =.008). Multivariate analysis showed that VEGF was not independent of the synthesis stage (P =.09). These data suggest that VEGF plays a role in the biology of CML and that VEGF inhibitors should be investigated in CML. PMID- 11877312 TI - Missense mutations of the WASP gene cause intermittent X-linked thrombocytopenia. AB - Mutations of the WASP gene have been previously shown to be responsible for classical Wiskott-Aldrich syndrome, isolated X-linked thrombocytopenia, and severe, congenital X-linked neutropenia. We report herewith 2 families in which affected males had a history of intermittent thrombocytopenia with consistently reduced platelet volume, in the absence of other major clinical features, and carried missense mutations of the WASP gene that allowed substantial protein expression. This observation broadens the spectrum of clinical phenotypes associated with WASP gene defects, and it indicates the need for molecular analysis in males with reduced platelet volume, regardless of the platelet number. PMID- 11877314 TI - Effect of SEA0400, a novel inhibitor of sodium-calcium exchanger, on myocardial ionic currents. AB - The effects of 2-[4-[(2,5-difluorophenyl) methoxy]phenoxy]-5-ethoxyaniline (SEA0400), a newly synthesized Na(+)-Ca(2+) exchanger (NCX) inhibitor, on the NCX current and other membrane currents were examined in isolated guinea-pig ventricular myocytes and compared with those of 2-[2-[4-(4-nitrobenzyloxy) phenyl]ethyl]isothiourea (KB-R7943). SEA0400 concentration-dependently inhibited the NCX current with a 10 fold higher potency than that of KB-R7943; 1 microM SEA0400 and 10 microM KB-R7943 inhibited the NCX current by more than 80%. KB R7943, at 10 microM, inhibited the sodium current, L-type calcium current, delayed rectifier potassium current and inwardly rectifying potassium current by more than 50%, but SEA0400 (1 microM) had no significant effect on these currents. These results indicate that SEA0400 is a potent and highly selective inhibitor of NCX, and would be a powerful tool for further studies on the role of NCX in the heart and the therapeutic potential of its inhibition. PMID- 11877315 TI - Proteinase-activated receptor-1 agonists attenuate nociception in response to noxious stimuli. AB - Proteinase-activated receptor-1 (PAR-1) is activated by thrombin and can be selectively activated by synthetic peptides (PAR-1-activating peptide: PAR-1-AP) corresponding to the receptor's tethered ligand. PAR-1 being expressed by afferent neurons, we investigated the effects of PAR-1 agonists on nociceptive responses to mechanical and thermal noxious stimuli. Intraplantar injection of selective PAR-1-AP increased nociceptive threshold and withdrawal latency, leading to mechanical and thermal analgesia, while control peptide had no effect. Intraplantar injection of thrombin also showed analgesic properties in response to mechanical, but not to thermal stimulus. Co-injection of PAR-1-AP with carrageenan significantly reduced carrageenan-induced mechanical and thermal hyperalgesia, while thrombin reduced carrageenan-induced mechanical but not thermal hyperalgesia. The fact that thrombin is not a selective agonist for PAR-1 may explain the different effects of thrombin and PAR-1-AP. These results identified analgesic properties for selective PAR-1 agonists that can modulate nociceptive response to noxious stimuli in normal and inflammatory conditions. PMID- 11877316 TI - The role of migrating leukocytes in IL-1 beta-induced up-regulation of kinin B(1) receptors in rats. AB - 1. The present study examines the role of migrating leukocytes in the ability of IL-1 beta to induce the functional up-regulation of B(1) receptors, as assessed by kinin B(1) agonist-induced oedema in the rat paw. 2. Pre-treatment with the PAF receptor antagonist WEB 2086 inhibited des-Arg(9)-BK-induced oedema in IL-1 beta-treated paws, while the LTB(4) receptor antagonist CP105696 had no effect. Des-Arg(9)-BK-induced paw oedema was also inhibited by pre-treatment with the selectin blocker fucoidin or by an anti-CD-18 monoclonal antibody. 3. I.d. injection of IL-1 beta produced a 5 - 10-fold increase of myeloperoxidase (MPO) activity in the rat paw. The increase in MPO activity was significantly inhibited by WEB 2086 (46 +/- 9%), fucoidin (68 +/- 5%) or the CD-18 antibody (84 +/- 3%). In contrast, i.d. injection of TNF alpha a dose known to upregulate the B(1) receptor functionally did not induce any significant increase in MPO activity. 4. Des-Arg(9)-BK alone had no effect in MPO activity but enhanced (by about 40%) the response induced by IL-1 beta, an effect prevented by the B(1) receptor antagonist des-Arg(9)-[Leu(8)]-BK. 5. The concentration of TNF-alpha was increased in the paws after i.d. injection of IL-1 beta. Pre-treatment with fucoidin, WEB 2086, anti-CD-18 or CP 105695, significantly reversed the local increases in TNF-alpha concentrations (80 +/- 2; 75 +/- 4, 73 +/- 3 and 40 +/- 2%), respectively. 6. Finally, IL-1 beta induced an increase of B(1) receptor mRNA levels in the rat paw, an effect which was prevented by fucoidin treatment. 7. Taken together, these results indicate that up-regulation of B(1) receptors in the rat paw following IL-1 beta seems to involve the local recruitment of neutrophils and subsequent local TNF-alpha production. The cross-talk between kinins, cytokines and leukocytes implicate B(1) receptors in chronic inflammatory diseases. PMID- 11877313 TI - NO as a signalling molecule in the nervous system. PMID- 11877317 TI - Chronic fluoxetine treatment selectively uncouples raphe 5-HT(1A) receptors as measured by [(35)S]-GTP gamma S autoradiography. AB - 1. Selective Serotonin Reuptake Inhibitors (SSRIs) are thought to have a delay in therapeutic efficacy because of the need to overcome the inhibitory influence of raphe 5-HT(1A) autoreceptors. Prolonged SSRI administration has been reported to desensitize these autoreceptors. We have used [(35)S]-GTP gamma S autoradiography to determine whether this desensitization occurs at the level of receptor/G protein coupling. 2. Male mice were injected intraperitoneally once a day with saline or 20 mg kg(-1) fluoxetine for either 2 days or 14 days. 5-HT(1A) receptor binding and coupling to G proteins were assessed using [(3)H]-8-OH-DPAT and [(35)S]-GTP gamma S autoradiography, respectively. 3. The 5-HT receptor agonist 5 carboxamidotryptamine (5-CT) stimulated [(35)S]-GTP gamma S binding in the substantia nigra, as well as in hippocampus and dorsal raphe nucleus. The 5 HT(1A) receptor antagonist p-MPPF (4-fluoro-N-(2-[4-(2-methoxyphenyl)1 piperazinyl]ethyl)-N-(2-pyridinyl)benzamide) blocked this effect in the latter regions, whereas the 5-HT(1B/D) antagonist GR-127,935 (2'-methyl-4'-(5-methyl [1,2,4]oxadiazol-3-yl)-biphenyl-4-carboxylic acid [4-methoxy-3-(4-methyl piperazin-l-yl)-phenyl]-amide) only decreased labelling in substantia nigra. 4. Fourteen-day fluoxetine treatment decreased 5-CT-stimulated [(35)S]-GTP gamma S binding in dorsal raphe (saline: 112 +/- 12% stimulation; fluoxetine: 66 +/- 13%), but not in substantia nigra (99 +/- 14% vs 103 +/- 7%) or hippocampus (157 +/- 3% vs 148 +/- 18%). Two-day fluoxetine treatment did not alter 5-CT stimulated [(35)S]-GTP gamma S binding in any of the brain areas investigated. 5. Decreased [(35)S]-GTP gamma S binding was not due to receptor down-regulation, since the density of raphe [(3)H]-8-OH-DPAT binding sites was unaffected by fluoxetine treatment. 6. These results suggest that the desensitization of presynaptic 5-HT(1A) receptor function occurs at the level of receptor-G protein interaction on dorsal raphe neurons, and may underlie the therapeutic efficacy of long-term SSRI treatment. PMID- 11877318 TI - Mechanisms of action of proteinase-activated receptor agonists on human platelets. AB - 1. We studied the activation of human platelets by thrombin and proteinase activated receptor (PAR)-activating peptides (PAR-APs) [SFLLRNPNDKYEPF-amide (TRAP), TFLLR-amide (PAR1AP) and AYPGKF-amide (PAR4AP)]. 2. PAR agonist-induced platelet aggregation, glycoprotein (GP) Ib and GPIIb/IIIa surface expression and ADP release were measured by light aggregometry, flow cytometry and chemiluminescence. 3. Aggregation inhibitors, including prostacyclin (PGI(2)), nitric oxide-releasing agent (S-nitroso-glutathione, GSNO), aspirin, apyrase, and phenanthroline were used to study the susceptibility of PAR agonist-induced aggregation to pharmacological inhibition. 4. Thrombin was the most potent platelet agonist, followed by PAR1AP, TRAP and PAR4AP. 5. The aggregatory potencies of PAR-APs were not modified by the aminopeptidase inhibitor, amastatin. 6. Subthreshold concentrations of PAR1AP potentiated the effects of PAR4AP to stimulate maximal aggregation. 7. Both PGI(2) and GSNO reduced PAR agonist-induced aggregation and diminished GPIIb/IIIa up-regulation. 8. PAR agonist-induced aggregation was aspirin-insensitive indicating a minor role for TXA(2). 9. In contrast, phenanthroline and apyrase significantly enhanced the anti-aggregatory effects of aspirin against thrombin-, PAR1AP- and TRAP-induced aggregation suggesting the involvement of ADP- and MMP-2-dependent pathways. 10. PAR4AP-induced aggregation (but not PAR1AP-induced aggregation) was entirely ADP dependent (abolished by apyrase) and resistant to phenanthroline (MMP-2 independent). 11. Thus, the mechanisms of PAR1 and 4-induced platelet aggregation are distinct and depend differentially on their ability to interact with pathways of aggregation, along with the subsequent activation of GPIIb/IIIa receptors. PMID- 11877319 TI - Characterization of an apamin-sensitive small-conductance Ca(2+)-activated K(+) channel in porcine coronary artery endothelium: relevance to EDHF. AB - 1. The apamin-sensitive small-conductance Ca(2+)-activated K(+) channel (SK(Ca)) was characterized in porcine coronary arteries. 2. In intact arteries, 100 nM substance P and 600 microM 1-ethyl-2-benzimidazolinone (1-EBIO) produced endothelial cell hyperpolarizations (27.8 +/- 0.8 mV and 24.1 +/- 1.0 mV, respectively). Charybdotoxin (100 nM) abolished the 1-EBIO response but substance P continued to induce a hyperpolarization (25.8 +/- 0.3 mV). 3. In freshly isolated endothelial cells, outside-out patch recordings revealed a unitary K(+) conductance of 6.8 +/- 0.04 pS. The open-probability was increased by Ca(2+) and reduced by apamin (100 nM). Substance P activated an outward current under whole cell perforated-patch conditions and a component of this current (38%) was inhibited by apamin. A second conductance of 2.7 +/- 0.03 pS inhibited by d tubocurarine was observed infrequently. 4. Messenger RNA encoding the SK2 and SK3, but not the SK1, subunits of SK(Ca) was detected by RT - PCR in samples of endothelium. Western blotting indicated that SK3 protein was abundant in samples of endothelium compared to whole arteries. SK2 protein was present in whole artery nuclear fractions. 5. Immunofluorescent labelling confirmed that SK3 was highly expressed at the plasmalemma of endothelial cells and was not expressed in smooth muscle. SK2 was restricted to the peri-nuclear regions of both endothelial and smooth muscle cells. 6. In conclusion, the porcine coronary artery endothelium expresses an apamin-sensitive SK(Ca) containing the SK3 subunit. These channels are likely to confer all or part of the apamin-sensitive component of the endothelium-derived hyperpolarizing factor (EDHF) response. PMID- 11877320 TI - 5-HT(2B) receptors play a key role in mediating the excitatory effects of 5-HT in human colon in vitro. AB - 1. 5-Hydroxytryptamine (5-HT) is known to produce a number of different effects in the gastrointestinal tract of various species, and has been proposed to play a key role in a number of intestinal disorders in man, including irritable bowel syndrome (IBS), although the receptors involved have yet to be established. The aim of the present study was to investigate the distribution and function of 5 HT(2B) receptors in human colon, and to establish their possible role in the aetiology of IBS. 2. The distribution of 5-HT(2B) receptor mRNA and protein were investigated by quantitative RT - PCR, Western analysis and immunocytochemistry. High levels of both mRNA and protein for 5-HT(2B) receptors were found throughout the human gastrointestinal tract, and in particular in colon, where 5-HT(2B) receptors were found predominantly in the longitudinal and circular smooth muscle layers within the muscularis externa, and in the myenteric nerve plexus lying between these two layers. 3. Electrical field stimulation of longitudinal muscle preparations of human colon mounted in organ baths resulted in neuronally mediated contractile responses, that were significantly potentiated by application of 5-HT (up to 10(-7) M), with a pEC(50) of 8.2 +/- 0.1 (n=49 donors). The response to 5-HT was inhibited by a number of selective 5-HT(2B) receptor antagonists. 4. This study has shown for the first time that, in contrast to animal studies, the excitatory effects of 5-HT in human colon are mediated by 5-HT(2B) receptors. It is proposed that these receptors contribute to the putative 5-HT-induced colonic smooth muscle hypersensitivity associated with IBS. PMID- 11877321 TI - Effect of agmatine on locus coeruleus neuron activity: possible involvement of nitric oxide. AB - 1. To investigate whether agmatine (the proposed endogenous ligand for imidazoline receptors) controls locus coeruleus neuron activity and to elucidate its mechanism of action, we used single-unit extracellular recording techniques in anaesthetized rats. 2. Agmatine (10, 20 and 40 microg, i.c.v.) increased in a dose-related manner the firing rate of locus coeruleus neurons (maximal increase: 95 +/- 13% at 40 microg). 3. I(1)-imidazoline receptor ligands stimulate locus coeruleus neuron activity through an indirect mechanism originated in the paragigantocellularis nucleus via excitatory amino acids. However, neither electrolytic lesions of the paragigantocellularis nucleus nor pretreatment with the excitatory amino acid antagonist kynurenic acid (1 micromol, i.c.v.) modified agmatine effect (10 microg, i.c.v.). 4. After agmatine administration (20 microg, i.c.v.), dose-response curves for the effect of clonidine (0.625 - 10 microg kg( 1) i.v.) or morphine (0.3 - 4.8 mg kg(-1) i.v.) on locus coeruleus neurons were not different from those obtained in the control groups. 5. Pretreatment with the nitric oxide synthase inhibitors N(omega)-nitro-L-arginine (10 microg, i.c.v.) or N(omega)-nitro-L-arginine methyl ester (100 microg, i.c.v.) but not with the less active stereoisomer N(omega)-nitro-D-arginine methyl ester (100 microg, i.c.v.) completely blocked agmatine effect (10 and 40 microg, i.c.v.). 6. Similarly, when agmatine (20 pmoles) was applied into the locus coeruleus there was an increase that was blocked by N(omega)-nitro-L-arginine methyl ester (100 microg, i.c.v.) in the firing rate of the locus coeruleus neurons (maximal increase 53 +/- 11% and 14 +/- 10% before and after nitric oxide synthase inhibition, respectively). 7. This study demonstrates that agmatine stimulates the firing rate of locus coeruleus neurons via a nitric oxide synthase-dependent mechanism located in this nucleus. PMID- 11877322 TI - KMUP-1 relaxes rabbit corpus cavernosum smooth muscle in vitro and in vivo: involvement of cyclic GMP and K(+) channels. AB - 1. In isolated endothelium-intact or denuded rabbit corpus cavernosum preconstricted with phenylephrine, KMUP-1 (0.001 - 10 microM) caused a concentration-dependent relaxation. 2. This relaxation of KMUP-1 was attenuated by endothelium removed, high K(+) and pretreatments with a soluble guanylyl cyclase (sGC) inhibitor ODQ (1 microM), a NOS inhibitor L-NAME (100 microM), a K(+) channel blocker TEA (10 mM), a K(ATP) channel blocker glibenclamide (1 microM), a voltage-dependent K(+) channel blocker 4-AP (100 microM) and Ca(2+) dependent K(+) channel blockers apamin (1 microM) and charybdotoxin (ChTX, 0.1 microM). 3. The relaxant responses of KMUP-1 (0.01, 0.05, 0.1 microM) together with a PDE inhibitor IBMX (0.5 microM) had additive actions on rabbit corpus cavernosum smooth muscle (CCSM). 4. KMUP-1 (0.01 - 10 microM) induced increase of intracellular cyclic GMP level in the primary cell culture of rabbit CCSM. This increase in cyclic GMP content was abolished in the presence of ODQ (10 microM). 5. Both KMUP-1 and sildenafil at 0.2, 0.4, 0.6 mg kg(-1) caused increases of intracavernous pressure (ICP) and duration of tumescene (DT) in a dose-dependent manner. These in vivo activities of ICP for sildenafil and KMUP-1 are consistent with those of in vitro effects of cyclic GMP. 6. KMUP-1 has the following merits: (1) inhibition of PDE or cyclic GMP breakdown, (2) stimulation of NO/sGC/cyclic GMP pathway, and (3) subsequent stimulation of K(+) channels, in rabbit CCSM. We suggest that these merits play prominent roles in KMUP-1-induced CCSM relaxation associated increases of ICP and penile erection. PMID- 11877323 TI - Extracellular signal-regulated kinase plays an essential role in endothelin-1 induced homotypic adhesion of human neutrophil granulocytes. AB - 1. Endothelin-1 (ET-1) stimulates integrin-dependent adhesion of neutrophil granulocytes to endothelial cells, one of the early key events in acute inflammation. However, the signalling pathway(s) of ET-1-stimulated neutrophil adhesive responses has not been elucidated. Previous studies indicated that extracellular signal-regulated kinase (ERK) activation could mediate rapid responses of neutrophil granulocytes to various stimuli. In this study, we investigated the role of ERK signalling in human neutrophil granulocytes challenged with ET-1. 2. ET-1 rapidly down-regulated the expression of L-selectin and up-regulated the expression of CD11b/CD18 on the neutrophil surface. Concomitantly, ET-1 induced homotypic adhesion (aggregation) of neutrophils, that was blocked by a monoclonal antibody to CD18. 3. ET-1, through ET(A) receptors, evoked activation of Ras and subsequent phosphorylation of Raf-1, mitogen activated protein kinase kinase (MAPK/ERK kinase) and ERK 1/2. ERK activation by ET-1 was rapid, concordant with the kinetics of ET-1-stimulated neutrophil aggregation. 4. Neutrophil responses to ET-1 were markedly attenuated by the MAPK/ERK kinase inhibitor PD98059, whereas inhibitors of p38 MAPK, tyrosine kinases and phosphatidylinositol 3-kinase had no detectable effects. We have observed a tight correlation between neutrophil ERK activation and homotypic adhesion. 5. These data indicate an essential role for ERK in mediating ET-1 stimulated adhesive responses of human neutrophil granulocytes. PMID- 11877324 TI - Probucol preserves endothelial function by reduction of the endogenous nitric oxide synthase inhibitor level. AB - 1. Oxide low-density lipoprotein (ox-LDL) is believed to play an important role in early events of atherogenesis, and asymmetric dimethylarginine (ADMA) is associated with the development of endothelial dysfunction. The present study examined the effect of a single injection of native low-density lipoprotein (LDL) on endothelium function and the serum level of ADMA and the effect of probucol on endothelium function and ADMA level in rats. 2. Endothelial injury was induced by intravenous injection of LDL at the dose of 2, 4, or 6 mg kg(-1) for 24, 48, or 72 h, and vasodilator responses to acetylcholine in the aortic rings and serum levels of ADMA, nitrite/nitrate (NO) and malondialdehyde (MDA) were determined. 3. Pretreatment with LDL markedly reduced endothelium-dependent relaxation in a concentration-dependent manner. Inhibition of vasodilator responses to acetylcholine by LDL was abolished in the presence of L-arginine (3 x 10(-4) M). Serum levels of ADMA and MDA were significantly elevated in the rats pretreated with LDL, while serum level of nitrite/nitrate was markedly decreased. 4. Pretreatment with probucol significantly improved endothelium-dependent relaxation, decreased concentrations of ADMA and MDA and increased nitrite/nitrate level in the rats treated with LDL. A similar effect was seen in the rats pretreated with an antioxidant vitamin E. 5. These results suggest that a single injection of native LDL causes endothelial dysfunction by elevation of ADMA levels and that the protective effect of probucol on endothelial cells is related to reduction of ADMA concentration. PMID- 11877325 TI - The flavonol quercetin activates basolateral K(+) channels in rat distal colon epithelium. AB - 1. The flavonol quercetin has been shown to activate a Cl(-) secretion in rat colon. Unlike the secretory activity of the related isoflavone genistein, quercetin's secretory activity does not depend on cyclic AMP; instead, it depends on Ca(2+). We investigated the possible involvement of Ca(2+) dependent basolateral K(+) channels using apically permeabilized rat distal colon epithelium mounted in Ussing chambers. 2. In intact epithelium, quercetin induced an increase in short-circuit current (I(sc)), which was diminished by the Cl(-) channel blockers NPPB and DPC, but not by glibenclamide, DIDS or anthracene-9 carboxylic acid. The effect of the flavonol was also inhibited by several serosally applied K(+) channel blockers (Ba(2+), quinine, clotrimazole, tetrapentylammonium, 293B), whereas other K(+) channel blockers failed to influence the quercetin-induced increase in I(sc) (tetraethylammonium, charybdotoxin). 3. The apical membrane was permeabilized by mucosal addition of nystatin and a serosally directed K(+) gradient was applied. The successful permeabilization was confirmed by experiments demonstrating the failure of bumetanide to inhibit the carbachol-induced current. 4. In apically permeabilized epithelium, quercetin induced a K(+) current (I(K)), which was neither influenced by ouabain nor by bumetanide. Whereas DPC, NPPB, charybdotoxin and 293B failed to inhibit this I(K), quinine, Ba(2+), clotrimazole and tetrapentylammonium were effective blockers of this current. 5. We conclude from these results that at least part of the quercetin-induced Cl(-) secretion can be explained by an activation of basolateral K(+) channels. PMID- 11877326 TI - Evidence of a novel site mediating anandamide-induced negative inotropic and coronary vasodilatator responses in rat isolated hearts. AB - 1. Cannabinoids are known to cause coronary vasodilatation and reduce left ventricular developed pressure (LVDP) in isolated hearts although the identity of the receptor(s) mediating these responses is unknown. Our objective was to pharmacologically characterize cannabinoid receptors mediating cardiac responses to the endocannabinoid, anandamide. 2. Dose-response curves for coronary perfusion pressure (CPP) and LVDP were constructed to anandamide, R-(+) methanandamide, palmitoylethanolamide (PEA) and JWH015 in isolated Langendorff perfused rat hearts. Anandamide dose-response curves were also constructed in the presence of antagonists selective for CB(1), CB(2) or VR(1) receptors. 3. Anandamide and methanadamide significantly reduced CPP and LVDP but the selective CB(2) receptor agonists, PEA and JWH015 had no significant effect, compared with equivalent vehicle doses. 4. Single bolus additions of the selective CB(1) receptor agonist, ACEA (5 nmol), decreased LVDP and CPP. When combined with JWH015 (5 nmol) these responses were not augmented. 5. Anandamide-mediated reductions in CPP were significantly blocked by the selective CB(1) receptor antagonists SR 141716A (1 microM) and AM251 (1 microM) and the selective CB(2) receptor antagonist SR 144528 (1 microM) but not by another selective CB(2) receptor antagonist AM630 (10 microM) nor the vanilloid VR(1) receptor antagonist capsazepine (10 microM). 6. SR 141716A, AM281 and SR 144528 significantly blocked negative inotropic responses to anandamide that were not significantly affected by AM251, AM630 and capsazepine. 7. One or more novel sites mediate negative inotropic and coronary vasodilatatory responses to anandamide. These sites can be distinguished from classical CB(1) and CB(2) receptors, as responses are sensitive to both SR 141716A and SR 144528. PMID- 11877327 TI - Ca(2+) signalling by recombinant human CXCR2 chemokine receptors is potentiated by P2Y nucleotide receptors in HEK cells. AB - 1. Human embryonic kidney (HEK)-293 cells expressing recombinant G alpha(i) coupled, human CXC chemokine receptor 2 (CXCR2) were used to study the elevation of the intracellular [Ca(2+)] ([Ca(2+)](i)) in response to interleukin-8 (IL-8) following pre-stimulation of endogenously expressed P2Y1 or P2Y2 nucleotide receptors. 2. Pre-stimulation of cells with adenosine 5'-triphosphate (ATP) revealed a substantial Ca(2+) signalling component mediated by IL-8 (E(max)=83 +/ 8% of maximal ATP response, pEC(50) of IL-8 response=9.7 +/- 0.1). 3. 1 microM 2 methylthioadenosine 5'-diphosphate (2MeSADP; P2Y1 selective) and 100 microM uridine 5'-triphosphate (UTP; P2Y2 selective) stimulated equivalent maximal increases in [Ca(2+)](i) elevation. However, UTP caused a sustained elevation, whilst following 2MeSADP [Ca(2+)](i) rapidly returned to basal levels. 4. Both UTP and 2MeSADP increased the potency and magnitude of IL-8-mediated [Ca(2+)](i) elevation but the effects of UTP (E(max) of IL-8 response increased to 50 +/- 1% of the maximal response to ATP, pEC(50) increased to 9.8 +/- 0.1) were greater than those of 2MeSADP (E(max) increased to 36 +/- 2%, pEC(50) increased to 8.7 +/ 0.2). 5. 5. The potentiation of IL-8-mediated Ca(2+) signalling by UTP was not dependent upon the time of IL-8 addition following UTP but was dependent on the continued presence of UTP. Potentiated IL-8 Ca(2+) signalling was apparent in the absence of extracellular Ca(2+), demonstrating the release of Ca(2+) from intracellular stores. 6. Activation of P2Y1 and P2Y2 receptors also revealed Ca(2+) signalling by an endogenously expressed, G alpha(s)-coupled beta adrenoceptor. 7. In conclusion, pre-stimulation of P2Y nucleotide receptors, particularly P2Y2, facilitates Ca(2+) signalling by either recombinant CXCR2 or endogenous beta-adrenoceptors. PMID- 11877328 TI - Modulation of contraction by alpha(2A/D)-adrenoceptors in mouse aorta: evidence employing knockout technology. AB - 1. We have investigated noradrenaline-evoked contractions in endothelium-denuded aorta from wild-type and alpha(2A/D)-adrenoceptor knockout mice. The maximum contraction to noradrenaline was significantly larger (1.36 +/- 0.24 mN, n=5) in aorta from knockout than from wild-type animals (0.78 +/- 0.14 mN, n=12, P<0.05), but there was no difference in potency of noradrenaline. There was no difference between groups in the contraction to KCl (80 mM) or PGF(2 alpha) (10 microM). 2. The contraction to noradrenaline (10 microM) was significantly larger in aorta from knockout animals, but yohimbine (1 microM) significantly increased this contraction (to 136 +/- 10% of control, n=6) in aorta from wild-type but not from knockout (97 +/- 10%, n=6, P<0.05). 3. In tissues precontracted with PGF(2 alpha) (10 microM), xylazine (1 microM) produced relaxations only in tissues from wild type mice. 4. The K(+) channel blocker glibenclamide (1 microM) had no significant effects on contractions to noradrenaline in either group. 5. It is concluded that an alpha(2A/D)-adrenoceptor exerts an inhibitory modulation of contraction in mouse aorta. PMID- 11877329 TI - Platelet-activating factor drives eotaxin production in an allergic pleurisy in mice. AB - 1. The activation of eosinophils via G-protein-coupled seven transmembrane receptors play a necessary role in the recruitment of these cells into tissue. The present study investigates a role for PAF in driving eotaxin production and eosinophil recruitment in an allergic pleurisy model in mice. 2. The intrapleural injection of increasing doses of PAF (10(-11) to 10(-9) moles per cavity) induced a dose- and PAF receptor-dependent recruitment of eosinophils 48 h after stimulation. 3. Intrapleural injection of PAF induced the rapid (within 1 h) release of eotaxin into the pleural cavity of mice and an anti-eotaxin antibody effectively inhibited PAF-induced recruitment of eosinophils. 4. Eosinophil recruitment in the allergic pleurisy was markedly inhibited by the PAF receptor antagonist UK-74,505 (modipafant, 1 mg kg(-1)). Moreover, recruitment of eosinophils in sensitized and challenged PAF receptor-deficient animals was lower than that observed in wild-type animals. 5. Blockade of PAF receptors with UK 74,505 suppressed by 85% the release of eotaxin in the allergic pleurisy. 6. Finally, the injection of a sub-threshold dose of PAF and eotaxin cooperated to induce eosinophil recruitment in vivo. 7. In conclusion, the production of PAF in an allergic reaction could function in multiple ways to facilitate the recruitment of eosinophils -- by facilitating eotaxin release and by cooperating with eotaxin to induce greater recruitment of eosinophils. PMID- 11877330 TI - Naloxone prevents cell-mediated immune alterations in adult mice following repeated mild stress in the neonatal period. AB - 1. Mild stress plus mild pain (solvent injection) applied daily to neonatal mice induces hormonal, behavioural and metabolic changes perduring in the adult life. 2. We investigated whether daily mild stress to neonatal mice induces also long term defined changes of immune response, and whether immune changes are prevented through repeated administration of the opioid antagonist naloxone. 3. Mild stress plus solvent injection administered from birth to the 21st postnatal day causes not only behavioural and metabolic changes, but also long-term (up to 110 days of life) splenocytes modifications, consisting in: increased release of the Th-1 type cytokines interleukin-2 (IL-2) (from an average of 346 to 788 pg ml(-1)), interferon-gamma (from 1770 to 3942) and tumour necrosis factor-alpha (from 760 to 1241); decreased release of the Th-2 type cytokines IL-4 (from 49.1 to 28.4) and IL-10 (from 1508 to 877). Moreover, enhanced natural killer-cell activity; enhanced proliferative splenocytes properties in resting conditions and following phytohemoagglutinin and concanavalin-A stimulation are observed. Immunological, behavioural and metabolic changes are prevented by the opioid antagonist ( )naloxone (1 mg kg(-1) per day s.c., administered instead of solvent) but not by the biologically inactive enantiomorph (+)naloxone. 4. In conclusion, endogenous opioid systems sensitive to naloxone are involved in long-lasting enhancement of the Th-1 type cytokines and cell-mediated immunological response caused by repeated mild stress administered postnatally. PMID- 11877331 TI - Characterization of the discriminable stimulus produced by 2-BFI: effects of imidazoline I(2)-site ligands, MAOIs, beta-carbolines, agmatine and ibogaine. AB - 1. The molecular nature and functions of the I(2) subtype of imidazoline binding sites are unknown but evidence suggests an association with monoamine oxidase (MAO). Rats can distinguish the selective imidazoline I(2)-site ligand 2-BFI from vehicle in drug discrimination, indicating functional consequences of occupation of these sites. We have used drug discrimination to investigate the nature of the discriminable stimulus, especially in relation to MAO inhibition. 2. Following training to distinguish 2-BFI 7 mg kg(-1) i.p. from saline vehicle in two-lever operant-chambers, male Hooded Lister rats underwent sessions where test substances were given instead and the proportion of lever presses on the 2-BFI associated lever (substitution) recorded. 3. 2-BFI; its cogeners BU216, BU224, BU226 and LSL60101; the reversible MAO-A inhibitors moclobemide and RO41-1049; the beta-carbolines harmane, norharmane and harmaline which also reversibly inhibit MAO-A, and the anti-addictive substance ibogaine exhibited potent, dose dependent substitution for 2-BFI. 4. Agmatine, and LSL60125 substituted at one dose only. The reversible MAO-B inhibitors lazabemide and RO16-1649; the sigma(2) site ligand SKF10,047 and the I(2A)-site ligand, amiloride, failed to substitute. The irreversible inhibitor of MAO, deprenyl, substituted for 2-BFI while clorgyline did not. 5. These results suggest imidazoline I(2) site ligands produce a common discriminable stimulus that appears associated with reversible inhibition of MAO-A rather than MAO-B, possibly through increases in extracellular concentration of one or more monoamines. Ibogaine exhibits a commonality in its subjective effects with those of I(2)-site ligands. PMID- 11877332 TI - Enhancement of 1 alpha,25-dihydroxyvitamin D(3)-induced differentiation of human leukaemia HL-60 cells into monocytes by parthenolide via inhibition of NF-kappa B activity. AB - 1. Transcription factors such as NF-kappa B provide powerful targets for drugs to use in the treatment of cancer. In this report parthenolide (PT), a sesquiterpene lactone of herbal remedies such as feverfew (Tanacetum parthenium) with NF-kappa B inhibitory activity, markedly increased the degree of human leukaemia HL-60 cell differentiation when simultaneously combined with 5 nM 1 alpha,25 dihydroxyvitamin D(3) (1,25-(OH)(2)D(3)). PT by itself did not induce HL-60 cell differentiation. 2. Cytofluorometric analysis indicated that PT stimulated 1,25 (OH)(2)D(3)-induced differentiation of HL-60 cells predominantly into monocytes. 3. Pretreatment of HL-60 cells with PT before the 1,25-(OH)(2)D(3) addition also potentiated the 1,25-(OH)(2)D(3)-induced HL-60 cell differentiation in both a dose- and a time-dependent manner, in which the enhanced levels of cell differentiation closely correlated with the inhibitory levels of NF-kappa B binding activity by PT. 4. In contrast, santonin, a sesquiterpene lactone without an inhibitory activity of NF-kappa B binding to the kappa B sites, did not enhance the 1,25-(OH)(2)D(3)-induced HL-60 cell differentiation. 5. In transfection experiments, PT enhanced 1,25-(OH)(2)D(3)-induced VDRE-dependent promoter activity. Furthermore, PT restored 1,25-(OH)(2)D(3)-induced VDRE dependent promoter activity inhibited by TNF-alpha, an activator of NF-kappa B signalling pathway. 6. These results indicate that PT strongly potentiates the 1,25-(OH)(2)D(3)-induced HL-60 cell differentiation into monocytes via the inhibition of NF-kappa B activity and provide evidence that inhibition of NF kappa B activation can be a pre-requisite to the efficient entry of promyelocytic leukaemia cells into a differentiation pathway. PMID- 11877333 TI - Characterization of pre- and postsynaptic muscarinic receptors in circular muscle of pig gastric fundus. AB - 1. This study investigated the subtype of muscarinic receptors on the cholinergic neurones and smooth muscle in the circular muscle of the pig gastric fundus. 2. Muscarinic antagonists, except MT-3, concentration-dependently inhibited the contractions induced by a given concentration of acetylcholine. Concentration response curves by acetylcholine were shifted rightwards in a parallel manner without depression of the maximum by the muscarinic antagonists, except by MT-3 that induced a leftward shift. Correlation of the pIC(50) and pA(2) values with published pK(i) values for the five muscarinic receptor subtypes suggests that the muscarinic receptors on pig gastric fundus circular muscle belong to the M(3) subtype. 3. Electrically-evoked contractions (40 V, 4 Hz, 0.25 ms, 2 min) were concentration-dependently inhibited by the muscarinic antagonists except for methoctramine and AF-DX 116, that increased the amplitude of the electrically induced contractions in lower concentrations. MT-3 tended to increase the electrically-induced contractions. 4. The antagonists, except MT-3, concentration dependently increased the electrically-induced tritium outflow (40 V, 4 Hz, 0.25 ms, 2 min) after incubation of the tissues with [(3)H]-choline. MT-3 (3 x 10(-8) and 10(-7) M) decreased the electrically-induced tritium release. Correlation of the pIC(50) values with published pK(i) values for the different muscarinic receptor subtypes yielded a significant and comparable correlation for M(1), M(3), M(4) and M(5) receptors. 5. These results suggest that the postsynaptic receptors in circular muscle of the pig gastric fundus belong to the M(3) subtype. However, the presynaptic receptor could not be clearly defined, although it does certainly not belong to the M(2) subtype. PMID- 11877334 TI - Systemic pre-treatment with a group II mGlu agonist, LY379268, reduces hyperalgesia in vivo. AB - 1. Previous studies investigating the role of metabotropic glutamate (mGlu) receptors in nociceptive processing have been hampered by the lack of systemically active, selective, ligands. This study investigates the possible analgesic and/or anti-hyperalgesic properties of the most potent compound to date that has systemic agonist activity at group II mGlu receptors, LY379268. 2. In testing the drug in rats as an analgesic to acute noxious stimuli, LY379268 (in doses up to 3 mg kg(-1) i.p.) did not affect withdrawal latencies to either mechanical or thermal stimulation. 3. However, when a 3 mg kg(-1) dose was given prior to an intraplantar injection of carrageenan, the inflammatory hyperalgesia that developed was significantly delayed compared to saline pre-treated controls, without affecting the inflammation of the paw. A similar dose of the mGlu inactive enantiomer, LY379267, was not anti-hyperalgesic. 4. In a model of mouse tail withdrawal to warm water, LY379268 (12 mg kg(-1) i.p.), given before a subcutaneous tail injection of capsaicin, reduced the subsequent neurogenic hyperalgesia. 5. Rota-rod testing showed that the drug did not produce a motor impairment in rats at antihyperalgesic doses. 6. The results indicate that systemic activation of this group of mGlu receptors reduces both inflammatory and neurogenic thermal hyperalgesia. PMID- 11877335 TI - Multiple pathways for fluoroquinolone secretion by human intestinal epithelial (Caco-2) cells. AB - 1. Human intestinal epithelial Caco-2 cells, T84 cells, and MDCKII cells transfected with human MDR1, were used to investigate the mechanistic basis of transintestinal fluoroquinolone secretion. 2. The fluoroquinolone grepafloxacin was secreted across Caco-2 monolayers by a saturable process (V(max)=16.9 +/- 3.4 nmol.cm(-2).h(-1)). Net secretion was reduced by 2-deoxyglucose/azide treatment to reduce intracellular ATP. 3. Grepafloxacin inhibited [(14)C]-ciprofloxacin (100 microM) secretion across Caco-2 monolayers (K(0.5)=0.8 mM), and concurrently increased the cellular accumulation of ciprofloxacin from the basal medium, indicating inhibition of export across the apical membrane. 4. The unconjugated bile acid, cholic acid, was secreted across Caco-2 monolayers, and this secretion was sensitive to inhibition by the MRP-selective inhibitor MK-571, suggesting MRP2 involvement. Secretion of cholic acid (10 microM) across the apical membrane was also inhibited by grepafloxacin (K(0.5)=0.3 mM), but not by ciprofloxacin. 5. In MDCKII-MDR1 monolayers, net secretion of grepafloxacin was increased by 3.5 fold compared with untransfected controls. Neither ciprofloxacin nor cholic acid showed net secretion in either MDCKII or MDCKII-MDR1 monolayers, showing that in contrast to grepafloxacin, neither are substrates for MDR1. 6. In T84 monolayers, which express MDR1 but not MRP2, neither ciprofloxacin nor cholic acid was secreted, whilst the V(max) for grepafloxacin secretion was lower than in Caco-2 cells, which express both MDR1 and MRP2. 7. In conclusion, the transepithelial secretion of grepafloxacin is mediated by both MRP2 and MDR1, whereas ciprofloxacin is a substrate for neither. Grepafloxacin also competes for the ciprofloxacin-sensitive pathway, which remains to be elucidated. PMID- 11877336 TI - The 5-series F(2)-isoprostanes possess no vasomotor effects in the rat thoracic aorta, the human internal mammary artery and the human saphenous vein. AB - 1. Among the F(2)-isoprostanes, the 15- and the 5-series are currently used as markers of lipid peroxidation in vascular diseases. 15-F(2t)-IsoP (also named iPF(2 alpha)-III) exerts a vasoconstriction in most vessels, whereas no data is available concerning 5-F(2t)-IsoP (also named iPF(2 alpha)-VI), which is more abundant in plasma. 2. The aim of this study was to determine whether 5-F(2t) IsoP possess any vascular effects on various vessels including the isolated rat thoracic aorta, the human internal mammary artery and the saphenous vein. 3. In organ baths, 5-F(2t)-IsoP and its 5-epimer did not affect the basal tone of any vessel, unlike 15-F(2t)-IsoP. These compounds possessed no antagonist effects on 15-F(2t)-IsoP-induced contractions, No dilator effect was observed in comparison with sodium nitroprusside and acetylcholine on the rat aorta. 4. In conclusion, we show that unlike 15-F(2t)-IsoP, 5-F(2t)-IsoP and its 5-epimer possess no vasomotor effects and as such are unlikely to be involved in the pathogenesis of vascular diseases. Further studies are required to test whether these mediators may have effects on systems not being measured in the current study. PMID- 11877338 TI - Protease-activated receptor-2 (PAR-2) in the rat gastric mucosa: immunolocalization and facilitation of pepsin/pepsinogen secretion. AB - 1. Agonists of protease-activated receptor-2 (PAR-2) trigger neurally mediated mucus secretion accompanied by mucosal cytoprotection in the stomach. The present study immunolocalized PAR-2 in the rat gastric mucosa and examined if PAR-2 could modulate pepsin/pepsinogen secretion in rats. 2. PAR-2-like immunoreactivity was abundant in the deep regions of gastric mucosa, especially in chief cells. 3. The PAR-2 agonist SLIGRL-NH(2), but not the control peptide LSIGRL-NH(2), administered i.v. repeatedly at 0.3 - 1 micromol kg(-1), four times in total, significantly facilitated gastric pepsin secretion, although a single dose produced no significant effect. 4. The PAR-2-mediated gastric pepsin secretion was resistant to omeprazole, N(G)-nitro-L-arginine methyl ester (L-NAME) or atropine, and also to ablation of sensory neurons by capsaicin. 5. Our study thus provides novel evidence that PAR-2 is localized in mucosal chief cells and facilitates gastric pepsin secretion in the rats, most probably by a direct mechanism. PMID- 11877337 TI - E-p-methoxycinnamic acid protects cultured neuronal cells against neurotoxicity induced by glutamate. AB - 1. We previously reported that four new phenylpropanoid glycosides and six known cinnamate derivatives isolated from roots of Scrophularia buergeriana Miquel (Scrophulariaceae) protected cultured cortical neurons from neurotoxicity induced by glutamate. Here, we have investigated the structure-activity relationships in the phenylpropanoids using our primary culture system. 2. The alpha,beta unsaturated ester moiety and the para-methoxy group in the phenylpropanoids appeared to play a vital role in neuroprotective activity. This suggested that E p-methoxycinnamic acid (E-p-MCA) might be a crucial component for their neuroprotective activity within the phenylpropanoid compounds. E-p-MCA significantly attenuated glutamate-induced neurotoxicity when added prior to an excitotoxic glutamate challenge. 3. The neuroprotective activity of E-p-MCA appeared to be more effective in protecting neurons against neurotoxicity induced by NMDA than from that induced by kainic acid. E-p-MCA inhibited the binding of [propyl-2,3-(3)H]-CGP39653 and [2-(3)H]-glycine to their respective binding sites on rat cortical membranes. However, even high concentrations of E-p-MCA failed to inhibit completely [propyl-2,3-(3)H]-CGP39653 and [2-(3)H]-glycine binding. 4. Indeed, E-p-MCA diminished the calcium influx that routinely accompanies glutamate-induced neurotoxicity, and inhibited the subsequent overproduction of nitric oxide and cellular peroxide in glutamate-injured neurons. 5. Thus, our results suggest that E-p-MCA exerts significant protective effects against neurodegeneration induced by glutamate in primary cultures of cortical neurons by an action suggestive of partial glutamatergic antagonism. PMID- 11877339 TI - Kainic acid-induced neuronal cell death in cerebellar granule cells is not prevented by caspase inhibitors. AB - 1. We examined the role of non-NMDA receptors in kainic acid (KA)-induced apoptosis in cultures of rat cerebellar granule cells (CGCs). KA (1 - 500 microM) induced cell death in a concentration-dependent manner, which was prevented by NBQX and GYKI 52466, non-NMDA receptor antagonists. Moreover, AMPA blocked KA induced excitotoxicity, through desensitization of AMPA receptors. 2. Similarly, KA raised the intracellular calcium concentration of CGCs, which was inhibited by NBQX and GYKI 52466. Again, AMPA (100 microM) abolished the KA (100 microM) induced increase in intracellular calcium concentration. 3. KA-induced cell death in CGCs had apoptotic features, which were determined morphologically, by DNA fragmentation, and by expression of the prostate apoptosis response-4 protein (Par-4). 5. KA (500 microM) slightly (18%) increased caspase-3 activity, which was strongly enhanced by colchicine (1 microM), an apoptotic stimulus. However, neither Z-VAD.fmk, a pan-caspase inhibitor, nor the more specific caspase-3 inhibitor, Ac-DEVD-CHO, prevented KA-induced cell death or apoptosis. In contrast, both drugs inhibited colchicine-induced apoptosis. 5. The calpain inhibitor ALLN had no effect on KA or colchicine-induced neurotoxicity. 6. Our findings indicate that colchicine-induced apoptosis in CGCs is mediated by caspase-3 activation, unlike KA-induced apoptosis. PMID- 11877340 TI - Permeability of porcine blood brain barrier to somatostatin analogues. AB - 1. Transport of a fluorescent somatostatin analogue (NBD-octreotide) across freshly isolated functionally intact capillaries from porcine brain was visualized by confocal microscopy and quantitated by image analysis. 2. Luminal accumulation of NBD-octreotide showed all characteristics of specific and energy dependent transport. Steady-state luminal fluorescence averaged 2 - 3 times cellular fluorescence and was reduced to cellular levels when metabolism was inhibited by NaCN. 3. The accumulation of NBD-octreotide in capillary lumens was inhibited in a concentration-dependent manner by unlabelled octreotide, by verapamil, PSC-833 and cyclosporin A, potent inhibitors of p-glycoprotein, and by leucotriene C(4), a strong modulator of Mrp2. Conversely, unlabelled octreotide reduced luminal accumulation of fluorescent BODIPY-verapamil on p-glycoprotein and of fluorescein-methotrexate, on Mrp2. None of the inhibitors used significantly reduced cellular accumulation of the fluorescent substrates. 4. Together, the data are consistent with octreotide being transported across the luminal membrane of porcine brain capillaries by both P-gp and Mrp2, providing further evidence that both transporters contribute substantially to the active barrier function of this endothelium. PMID- 11877341 TI - Allergenic sensitization prevents upregulation of haemopoiesis by cyclo-oxygenase inhibitors in mice. AB - 1. We evaluated whether immunization affects bone-marrow responses to indomethacin, because allergenic sensitization and challenge upregulate responses to haemopoietic cytokines (including IL-5-driven eosinopoiesis) in murine bone marrow, while indomethacin upregulates haemopoiesis and protects bone-marrow from radiation damage. 2. Progenitor (semi-solid) and/or precursor (liquid) cultures were established from bone-marrow of: (a) normal mice; (b) ovalbumin-sensitized mice, with or without intranasal challenge. Cultures were established with GM-CSF (2 ng ml(-1)) or IL-5 (1 ng ml(-1)), respectively, alone or associated with indomethacin (10(-7) - 10(-11) M) or aspirin (10(-7) - 10(-8) M). Total myeloid colony numbers and numbers of eosinophil-peroxidase-positive cells were determined at day 7. 3. In naive BALB/c mice, indomethacin (10(-7) - 10(-9) M) increased GM-CSF-stimulated myeloid colony formation (P=0.003 and P=0.009, respectively). In contrast, it had no effect on bone-marrow of ovalbumin sensitized and challenged mice. Indomethacin (10(-7) - 10(-9) M) also increased eosinophil precursor responses to IL-5 in bone-marrow of naive (P<0.001 and P=0.002 respectively), but not sensitized-challenged mice. Aspirin (10(-7) M) had similar effects, equally abolished by sensitization. Enhancement of haemopoiesis by indomethacin required adherent cells from naive bone-marrow. Nonadherent cells responded to IL-5 but not to indomethacin. Indomethacin was effective on bone marrow from sham-sensitized, ovalbumin-challenged, but not from sensitized, saline-challenged mice. Plasma transfer from immune mice abolished eosinophil precursor responses to indomethacin in bone-marrow of naive recipients. This was not prevented by previous removal of antibody from immune plasma. 4. COX inhibitors enhance haemopoiesis in naive but not allergic mice. Responsiveness to indomethacin can be abolished either by active sensitization or by immune plasma transfer. Specific antibody is not involved. PMID- 11877342 TI - Adenosine A(2A) receptors in portal hypertension: their role in the abnormal response to adenosine of the cranial mesenteric artery in rabbits. AB - 1. Adenosine is a regulator of mesenteric vasodilation involved in auto regulation and post-prandial hyperemia, but the adenosine receptor subtype involved in this relaxant effect is poorly characterized. We have now pharmacologically characterized this receptor in rabbit mesenteric arteries and investigated how this adenosine receptor response changes in portal hypertensive animals since the adenosine response is decreased. 2. The closest non metabolisable adenosine analogue, 2-chloroadenosine (CADO), the mixed A(1)/A(2) receptor agonist, 5'-ethylcarboxamidoadenosine (NECA), and the selective A(2A) receptor agonist, 2-[4-(2-p-carbonyethyl)phenylamino]-5'-N ethylcarboxamidoadenosine (CGS 21680) (1 pM -- 1 mM) relaxed noradrenaline pre contracted arteries with a rank order of potency of CGS 21680 (EC(50)=20 nM) > or = NECA (60 nM)>>CADO (640 nM). 3. The selective A(2A) receptor antagonist, 4-(2 [7-amino-2-(2-furyl)-[1,2,4]-triazolo[2,3-a][1,3,5]-triazin-5 ylamino]ethyl)phenol (ZM 241385, 100 nM), shifted to the right the CADO concentration-response curve. 4. In portal hypertensive animals, there was mainly a decreased potency but also a decreased efficacy of all tested adenosine agonists compared to normal animals. Concomitantly, there was a decreased adenosine plasma level and a decreased binding density of [(3)H]-CGS 21680 and [(3)H]-ZM 241385 to mesenteric artery membranes from portal hypertensive compared to normal rabbits. 5. These results indicate that A(2A) receptor activation is required for the adenosine-induced mesenteric relaxation and that the decreased density of A(2A) receptors may contribute to the decreased relaxation induced by adenosine of mesenteric arteries in portal hypertensive animals. PMID- 11877343 TI - Tachykinins via Tachykinin NK(2) receptor activation mediate ozone-induced increase in the permeability of the tracheal mucosa in guinea-pigs. AB - 1. Acute exposure to ozone is known to cause airway hyperresponsiveness, which, at least in part, seems to result from an increase in the permeability of the airway mucosa. Recently, we demonstrated that depletion of sensory neuropeptides inhibits the ozone-induced increase in the permeability of the tracheal mucosa in guinea-pigs. The aim of this study was to determine whether tachykinins mediate ozone-induced increase in the permeability of the tracheal mucosa in guinea-pigs. 2. Anaesthetized guinea-pigs were exposed to either 3 p.p.m. ozone or filtered air for 30 min. Immediately after exposure, a tracheal segment was isolated in vivo and administered with horseradish peroxidase (HRP). The permeability was assessed by monitoring the appearance of HRP in the blood. 3. A low dose of NKA increased the permeability of the tracheal mucosa, whereas a low dose of SP was without effect. Low and high doses of the selective NK(3) receptor agonist, senktide, were also without effect. The effect of a low dose of NKA was abolished by the NK(2) receptor antagonist, SR-48,968. A high dose of SP increased the permeability in a manner reversible by the NK(1) receptor antagonist, CP-96,345. 4. Pretreatment with SR-48,968 completely inhibited the ozone-induced increase in the permeability, whereas CP-96,345 had no effect. 5. It is thus concluded that endogenous tachykinins mediate the ozone-induced increase in the permeability of the tracheal mucosa in guinea-pigs mainly via NK(2) receptor activation. PMID- 11877344 TI - Algogen-specific pain processing in mouse spinal cord: differential involvement of voltage-dependent Ca(2+) channels in synaptic transmission. AB - 1. The effects of intrathecal (i.t.) administration of N-, P/Q- or L-type voltage dependent Ca(2+)-channel blockers were tested in two pain models involving bradykinin (BK)- and alpha,beta-methylene ATP (alpha,beta meATP)-induced activation of primary afferent neurons in mice. 2. The nociceptive response (amount of time spent licking and biting the hindpaw) induced by intraplantar injection of BK (500 pmol mouse(-1)) was significantly attenuated by both omega conotoxin GVIA (N-type blocker) and calciseptine (L-type) but not by omega agatoxin IVA (P/Q-type). 3. The nociceptive response induced in a similar way by alpha,beta meATP (100 nmol) was significantly inhibited by both the above N- and P/Q-type Ca(2+)-channel blockers but not by the L-type blocker. 4. The nociceptive responses elicited by BK and alpha,beta meATP were dose-dependently inhibited by a tachykinin-NK1-receptor antagonist (L-703,606) and an N-methyl-D aspartate (NMDA)-receptor antagonist (D-AP5), respectively. 5. Intrathecal administration of substance P (SP) (1.8 nmol) or NMDA (350 pmol) elicited algesic responses, such as licking, biting and scratching of the hindquarters. The SP induced algesic behaviour was significantly inhibited by the L-type blocker but not by the N-type. The NMDA-induced response was not affected by either the N- or the P/Q-type blocker. 6. These findings suggest that BK and ATP most likely excite different types of sensory neurons in the periphery and that within the spinal cord the former stimulates peptidergic transmission regulated by presynaptic N- and postsynaptic L-type Ca(2+) channels, while the latter stimulates glutamatergic transmission regulated by presynaptic N- and P/Q-type channels. PMID- 11877346 TI - Cardiology patient page. The implantable cardioverter-defibrillator: patient perspective. PMID- 11877347 TI - Peroxisome proliferator-activated receptor alpha as a genetic determinant of cardiac hypertrophic growth: culprit or innocent bystander? PMID- 11877348 TI - Increased prevalence of carotid atherosclerosis in hepatitis B virus carriers. AB - BACKGROUND: Recent experimental and epidemiological findings suggest that some infectious agents play a role in the development and promotion of atherosclerosis. We have investigated the possible association between hepatitis B virus surface antigen (HBsAg) positivity and carotid arteriosclerosis. METHODS AND RESULTS: In this cross-sectional cohort study, we analyzed data from subjects undergoing general health-screening tests, including both high-resolution B-mode carotid ultrasound and assessment of HBsAg status, between 1994 and 2001 at our institute. Of the 4686 study subjects (3137 men and 1549 women; age 22 to 88 years), 1294 (28%) had carotid artery plaque and 40 (0.9%) were positive for HBsAg, indicating they were hepatitis B virus carriers. No HBsAg-positive subjects were positive either for antibodies against the hepatitis C virus (HCV) or for HCV core proteins. Univariate analysis revealed HBsAg positivity was associated with carotid plaque with an odds ratio of 1.58 (95% CI, 1.14 to 2.19, P<0.05). When other confounding risk factors for atherosclerosis were included as covariates in the statistical analysis, HBsAg positivity was still positively associated with carotid plaque with an odds ratio of 1.57 (95% CI, 1.10 to 2.24, P<0.05). CONCLUSIONS: These findings suggest a possible role of chronic hepatitis B infection in the pathogenesis of carotid arteriosclerosis. PMID- 11877349 TI - Norepinephrine turnover is increased in suprabulbar subcortical brain regions and is related to whole-body sympathetic activity in human heart failure. AB - BACKGROUND: Although it is established that heightened sympathetic drive exists in congestive heart failure (CHF), the reflex processes by which this may occur and the sites in the central nervous system that may be responsible for mediating this process are not yet fully elucidated. METHODS AND RESULTS: Eight patients with moderate to severe CHF and 8 healthy control subjects underwent simultaneous arterial and bilateral internal jugular venous blood sampling and cerebral venous blood pool scanning for anatomical determination of the origin of internal jugular venous blood flow. We estimated sympathetic nervous activity by measuring total body norepinephrine (NE) spillover using radiotracer methodology and determined brain NE turnover by measuring the internal jugular overflow of NE and its lipophilic metabolites, 3-methoxy-4-hydroxyphenylglycol and 3,4 dihydroxyphenylglycol. Suprabulbar subcortical turnover of NE was significantly greater in CHF patients than in the healthy group (2.77 +/- 0.75 versus 0.66 +/- 0.40 nmol/min, P<0.05). There was a significant positive correlation between suprabulbar subcortical turnover of NE and total body NE spillover (r=0.62, P=0.01). CONCLUSIONS: This study, for the first time, demonstrates elevated suprabulbar subcortical noradrenergic activity in human CHF and identifies a positive correlation between this and the level of whole-body NE spillover. The findings suggest that the activation of noradrenergic neurons projecting rostrally from the brain stem mediates sympathetic nervous stimulation in CHF. PMID- 11877350 TI - Selective upregulation of endothelin B receptor gene expression in severe pulmonary hypertension. AB - BACKGROUND: The pulmonary circulation is an important site for the production and clearance of endothelin (ET)-1, a potent vasoactive and mitogenic peptide. Increased plasma ET-1 levels are observed in pulmonary arterial hypertension (PHT) and may contribute to the regulation of pulmonary vascular resistance, as well as to proliferative changes in the pulmonary vascular bed. METHODS AND RESULTS: We prospectively assessed changes in plasma big ET-1 levels and changes in ET(A) and ET(B) receptor gene expression in 14 consecutive patients undergoing pulmonary thromboendarterectomy for thromboembolic PHT. Plasma big ET-1 levels were higher in patients with PHT (median, 2.2 pg/mL; 25th to 75th percentile, 1.5 to 3.0 pg/mL) compared with age-matched controls (median, 1.2 pg/mL; 25th to 75th percentile, 1.0 to 1.4 pg/mL; P=0.002). In addition to increased plasma big ET-1 levels, selective upregulation of ET(B) receptor mRNA transcripts and immunoreactive protein in the pulmonary artery was observed in the patients; however, ET(A) receptor gene expression was unaffected. CONCLUSIONS: These data suggest that changes in the ET signaling system in PHT caused by thromboembolic disease are not limited to an increased production of ET-1: they also affect ET receptor gene expression. PMID- 11877351 TI - Homocysteine induces 3-hydroxy-3-methylglutaryl coenzyme a reductase in vascular endothelial cells: a mechanism for development of atherosclerosis? AB - BACKGROUND: It has been established that hyperhomocyst(e)inemia (HHCy) is an independent and graded risk factor for atherosclerosis, although the molecular link to the atherosclerotic process remains obscure. METHODS AND RESULTS: Screening human umbilical vein endothelial cells (HUVECs) with complementary DNA microarray for the gene expression modified by homocysteine (Hcy) revealed that 3 hydroxy-3-methylglutaryl coenzyme A reductase (HMGCR) was upregulated. This effect was confirmed using quantitative reverse transcriptase-polymerase chain reaction. Actinomycin D studies revealed that Hcy stabilized HMGCR mRNA (tau(1/2), 9.5 +/- 1.0 versus 5.0 +/- 0.2 hours). Expression of immunodetectable HMGCR in both HUVECs and renal microvascular endothelial cells was increased in Hcy-treated cells in association with the increased abundance of caveolin. Application of a cell-permeable superoxide dismutase mimetic, Mn-TBAP, reversed the Hcy-induced expression of HMGCR. Additional biochemical analysis of the abundance of total cellular cholesterol showed that 0, 20, 50, and 100 micromol/L Hcy resulted in 22.2 +/- 7.3%, 39.5 +/- 1.2%, and 50.4 +/- 6.8% increase, respectively. Gas chromatography mass spectrometry analysis of extracted cholesterol from Hcy-treated HUVECs and from the culture medium showed 17.8 +/- 5.2% and 24.0 +/- 14.5% increases, respectively. Application of simvastatin to Hcy-treated cells reduced cellular cholesterol and prevented Hcy-induced suppression of NO production by HUVECs in a dose-dependent manner. CONCLUSIONS: Using a cDNA microarray, the data disclosed an unexpected link between Hcy and cholesterol dysregulation based on the finding of increased abundance of HMGCR mRNA and protein in endothelial cells, demonstrated the possible role of Hcy induced oxidative stress in this response, and revealed the improvement of endothelial NO production in Hcy-treated HUVECs by statins. Collectively, these findings may provide a solid explanation for the observed proatherogenic effect of HHcy. PMID- 11877352 TI - Patients with Glanzmann thrombasthenia lacking platelet glycoprotein alpha(IIb)beta(3) (GPIIb/IIIa) and alpha(v)beta(3) receptors are not protected from atherosclerosis. AB - BACKGROUND: Platelets have been suggested to play a role in the early development of atherosclerosis. As one test of this hypothesis, we assessed whether patients with Glanzmann thrombasthenia who lack platelet glycoprotein alpha(IIb)beta(3) (GPIIb/IIIa) complexes or both alpha(IIb)beta(3) and the more ubiquitous alpha(v)beta(3) cell membrane complexes are protected from development of atherosclerosis. METHODS AND RESULTS: Seven patients with Glanzmann thrombasthenia, 45 to 66 years of age, underwent bilateral carotid artery ultrasonography and screening for risk factors of atherosclerosis. Findings consistent with early atherosclerosis evaluated by measurement of intima-media thickness and presence of atherosclerotic plaques were observed in 6 of the 7 patients. Intima-media thickness values higher than the 75th and 90th percentiles of age- and sex-matched white control subjects of the Atherosclerosis Risk in Communities (ARIC) study were observed in 30 and 8 of 56 carotid artery measurements, respectively. Five of the 6 patients with signs consistent with early atherosclerosis lacked both alpha(IIb)beta(3) and alpha(v)beta(3) complexes and 1 only lacked alpha(IIb)beta(3). CONCLUSIONS: Glanzmann thrombasthenia does not protect affected individuals from development of atherosclerosis. PMID- 11877353 TI - Five-year risk of cardiac mortality in relation to initial severity and one-year changes in depression symptoms after myocardial infarction. AB - BACKGROUND: Although previous research demonstrated an independent link between depression symptoms and cardiac mortality after myocardial infarction (MI), depression was assessed only once, and a dose-response relationship was not evaluated. METHODS AND RESULTS: We administered the Beck Depression Inventory to 896 post-MI patients during admission and at 1 year. Five-year survival was ascertained using Medicare data. We observed a significant long-term dose response relationship between depression symptoms during hospitalization and cardiac mortality. Results remained significant after control for multiple measures of cardiac disease severity. Although 1-year scores were also linked to cardiac mortality, most of that impact was explained by baseline scores. Improvement in depression symptoms was associated with less cardiac mortality only for patients with mild depression. Patients with higher initial scores had worse long-term prognosis regardless of symptom changes. CONCLUSIONS: The level of depression symptoms during admission for MI is more closely linked to long term survival than the level at 1 year, particularly in patients with moderate to severe levels of depression, suggesting that the presumed cardiovascular mechanisms linking depression to cardiac mortality may be more or less permanent for them. PMID- 11877354 TI - Mechanism of the blood pressure--raising effect of cocaine in humans. AB - BACKGROUND: Although the sympathomimetic actions and cardiovascular complications of cocaine are ascribed to inhibition of norepinephrine (NE) reuptake, this hypothesis has not been tested in humans. We asked (a) whether cocaine can inhibit NE reuptake in the human peripheral circulation and (b) whether the NE mediated peripheral vasoconstriction is the main mechanism mediating blood pressure-raising effect of cocaine. METHODS AND RESULTS: In 15 healthy cocaine naive subjects, we measured blood pressure, forearm blood flow, and forearm venous NE concentration during administration of (a) intrabrachial cocaine (0.15 and 15 mg), which produced no systemic neurohormonal effects, and (b) intranasal cocaine (2 mg/kg). Intrabrachial cocaine (0.15 mg) increased venous forearm NE concentration by 82% and vascular resistance by 71% (P<0.01). Increasing the intrabrachial cocaine dose by 100-fold to match the venous cocaine level of massive cocaine overdose caused a small additional increase in venous forearm NE concentration without causing significant additional vasoconstriction. Although intranasal cocaine (2 mg/kg) matched the venous cocaine concentrations caused by 0.15 mg of intrabrachial cocaine, venous NE concentration was unchanged as sympathetic nerve activity (SNA) decreased reflexively as the result of an increase in blood pressure. When SNA was restored to baseline by blunting the cocaine-induced rise in blood pressure (baroreflex activation) with nitroprusside, venous NE concentration increased to the same level caused by intrabrachial cocaine. CONCLUSIONS: In healthy cocaine-naive individuals, cocaine can inhibit NE reuptake in the human peripheral circulation. However, this mechanism does not contribute importantly to the blood pressure-raising effect of cocaine because activation of baroreceptor reflexes decreases SNA, the neural stimulus for NE release. PMID- 11877356 TI - Analysis of T-wave morphology from the 12-lead electrocardiogram for prediction of long-term prognosis in male US veterans. AB - BACKGROUND: The aim of the present study was to assess the prognostic value of novel repolarization descriptors from the 12-lead ECG in a large cohort of US veterans. METHODS AND RESULTS: Male US veterans (n=813) with cardiovascular disease had digital 12-lead ECGs recorded at the VA Medical Center, Washington, DC, between 1984 and 1991. The patient series was retrospectively compiled in 1991; follow-up was prospectively assessed until 2000. Novel ECG variables characterizing repolarization and the T-wave loop were automatically analyzed. Of 772 patients with technically analyzable data, 252 patients (32.6%) died after a mean follow-up of 10.4 +/- 3.8 years. Direct comparison between dead and alive patients showed that the so-called T-wave residua (the absolute and relative amount of nondipolar contents within the T wave) predicted mortality (111 900 +/- 164 700 versus 85 600 +/- 144 800 between dead and alive patients, P<0.0002; and 0.43 +/- 0.62% versus 0.33 +/- 0.56%, P<0.0005 for the absolute and relative T wave residuum, respectively). On Cox regression analysis entering age, left ventricular ejection fraction, echocardiographic left ventricular hypertrophy, and either of the T-wave residua, risk prediction was independent for the absolute (P=0.022) and for the relative (P=0.006) T-wave residuum, respectively, with age (P<0.0001), presence of left ventricular hypertrophy (P=0.002), and left ventricular ejection fraction (P=0.004) also being predictors of survival. CONCLUSIONS: The heterogeneity of myocardial repolarization, measured by the so called T-wave residuum in the ECG, confers long-term independent prognostic information in US veterans with cardiovascular disease. PMID- 11877355 TI - Validation of collateral fractional flow reserve by myocardial perfusion imaging. AB - BACKGROUND: Collateral fractional flow reserve (FFR(coll)) is an index to quantify collateral blood flow, derived from coronary pressure measurements. Although well defined theoretically, its direct validation by myocardial perfusion imaging has not been established so far. Validating this index by myocardial perfusion imaging is the main aim of this study. METHODS AND RESULTS: Twenty-four consecutive patients with stable angina and single left anterior descending artery stenosis underwent simultaneous measurement of aortic pressure (P(a)), coronary wedge pressure (P(w)), and central venous pressure (P(v)) during balloon inflation. FFR(coll) was calculated and compared with the extent and severity of the defect during coronary occlusion using (99m)Tc-sestamibi imaging at balloon inflation of the respective coronary artery. Although the pressure derived collateral indexes (P(w), P(w)/P(a), and FFR(coll)) ranged widely, they were closely correlated with extent and severity scores of the nuclear occlusion images and superior to the ECG for that purpose. Of all parameters, FFR(coll) correlated best with the severity score at imaging (r=-0.88), followed by the P(w)/P(a) ratio (r=-0.74) or P(w) alone (r=-0.69). CONCLUSIONS: FFR(coll), calculated from coronary pressure during balloon occlusion, is highly correlated with the extent and severity of the defect at myocardial perfusion of the territory of the occluded artery and can be used for quantitative assessment of collateral blood flow in conscious humans. PMID- 11877357 TI - Change in diastolic left ventricular filling after one year of antihypertensive treatment: The Losartan Intervention For Endpoint Reduction in Hypertension (LIFE) Study. AB - BACKGROUND: It is well established that hypertensive patients with left ventricular (LV) hypertrophy have impaired diastolic filling. However, the impact of antihypertensive treatment and LV mass reduction on LV diastolic filling remains unclear. METHODS AND RESULTS: Echocardiograms were recorded in 728 hypertensive patients with ECG-verified LV hypertrophy (Cornell voltage-duration or Sokolow-Lyon) at baseline and after 1 year of blinded treatment with either losartan or atenolol-based regimen. Systolic and diastolic blood pressures (BP) were reduced on average 23/11 mm Hg; isovolumic relaxation time and E/A ratio became more normal, and LV inflow deceleration time prolonged (all P<0.001). Directionally opposite changes in isovolumic relaxation time (IVRT) and deceleration time indicate improvement in active LV relaxation and passive chamber stiffness during early diastole. Prevalences of normal LV filling increased, abnormal relaxation and pseudonormalization decreased, and restrictive filling pattern remained unchanged (P<0.05). Patients with reduction in LV mass had smaller left atrial diameter, shortened IVRT, increased E/A ratio, and prolonged LV inflow deceleration time (all P<0.001). Patients without LV mass reduction had no change in diastolic filling parameters (P=NS). IVRT shortening was independently associated with reduction in LV mass. Increase in E/A ratio was independently associated with reduction in diastolic BP, and increase in the deceleration time was independently associated with reduced end-systolic relative wall thickness. CONCLUSIONS: Antihypertensive therapy resulting in LV mass or relative wall thickness regression is associated with significant improvement of diastolic filling parameters related to active relaxation and passive chamber stiffness compared with patients without regression, independent of BP reduction; however, abnormalities of diastolic LV filling remain common. PMID- 11877358 TI - Pulmonary vein isolation for paroxysmal and persistent atrial fibrillation. AB - BACKGROUND: The pulmonary veins (PVs) have been demonstrated to often play an important role in generating atrial fibrillation (AF). The purpose of this study was to determine the safety and efficacy of segmental PV isolation in patients with paroxysmal or persistent AF. METHODS AND RESULTS: In 70 consecutive patients (mean age, 53 +/- 11 years) with paroxysmal (58) or persistent (12) AF, segmental PV isolation guided by ostial PV potentials was performed. The left superior, left inferior, and right superior PVs were targeted for isolation in all patients, and the right inferior PV was isolated in 20 patients. Among the 230 targeted PVs, 217 (94%) were completely isolated, with a mean of 6.5 +/- 4.2 minutes of radiofrequency energy applied at a maximum power setting of 35 W. A second PV isolation procedure was performed in 6 patients (9%). At 5 months of follow-up, 70% of patients with paroxysmal and 22% of patients with persistent AF were free from recurrent AF (P<0.001), and 83% of patients with paroxysmal AF were either free of symptomatic AF or had significant improvement. Among various clinical characteristics, only paroxysmal AF was an independent predictor of freedom from recurrence of AF (P<0.05). One patient developed unilateral quadrantopsia after the procedure. There were no other complications. CONCLUSIONS: With a segmental isolation approach that targets at least 3 PVs, a clinically satisfactory result can be achieved in >80% of patients with paroxysmal AF. The clinical efficacy of pulmonary vein isolation is much lower when AF is persistent than when it is paroxysmal. PMID- 11877359 TI - Risk and predictors of stroke after myocardial infarction among the elderly: results from the Cooperative Cardiovascular Project. AB - BACKGROUND: Stroke is an important outcome after acute myocardial infarction. Studies that have examined this relationship have largely excluded older patients, even though half of stroke admissions occur among patients 75 years of age and older. METHODS AND RESULTS: Among 111 023 medicare patients discharged with a principal diagnosis of acute myocardial infarction during an 8-month period in 1994 to 1995, we identified hospital admissions for ischemic stroke within 6 months of discharge. The rate of admission was 2.5% within 6 months. Independent predictors of ischemic stroke were age greater-than-or-equal 75 years, black race, no aspirin at discharge, frailty, prior stroke, atrial fibrillation, diabetes, hypertension, and history of peripheral vascular disease. To identify individuals at increased risk for stroke, a risk stratification score was constructed from identified factors. The 6-month stroke admission rate for patients with a score of 4 or higher (approximately 20% of the total sample) was approximately 4%. CONCLUSIONS: The risk of stroke after myocardial infarction is substantial, with about 1 in 40 patients suffering an ischemic stroke within 6 months of discharge. Simple clinical factors can predict the risk of stroke and, based on these factors, we identified 20% of older patients who have a 1 in 25 chance of being hospitalized for a stroke within 6 months of discharge. PMID- 11877360 TI - Fetal, infant, and childhood growth and adult blood pressure: a longitudinal study from birth to 22 years of age. AB - BACKGROUND: People who are small at birth tend to have higher blood pressure in later life. However, it is not clear whether it is fetal growth restriction or the accelerated postnatal growth that often follows it that leads to higher blood pressure. METHODS AND RESULTS: We studied blood pressure in 346 British men and women aged 22 years whose size had been measured at birth and for the first 10 years of life. Their childhood growth was characterized using a conditional method that, free from the effect of regression to the mean, estimated catch-up growth. People who had been small at birth but who gained weight rapidly during early childhood (1 to 5 years) had the highest adult blood pressures. Systolic pressure increased by 1.3 mm Hg (95% CI, 0.3 to 2.3) for every standard deviation score decrease in birth weight and, independently, increased by 1.6 mm Hg (95% CI, 0.6 to 2.7) for every standard deviation score increase in early childhood weight gain. Adjustment for adult body mass index attenuated the effect of early childhood weight gain but not of birth weight. Relationships were smaller for diastolic pressure. Weight gain in the first year of life did not influence adult blood pressure. CONCLUSIONS: Part of the risk of adult hypertension is set in fetal life. Accelerated weight gain in early childhood adds to this risk, which is partly mediated through the prediction of adult fatness. The primary prevention of hypertension may depend on strategies that promote fetal growth and reduce childhood obesity. PMID- 11877361 TI - Trunk fat and blood pressure in children through puberty. AB - BACKGROUND: Fat distribution is well recognized as a cardiovascular risk factor in adults. The association between android fat distribution and cardiovascular risk factors, such as blood pressure (BP), was previously reported in an African American and Caucasian pediatric population. The aim of the present study was to investigate the relationship between BP and body fat distribution in a large cross-sectional pediatric sample. The effects of race, sex, and puberty on this relationship were assessed. METHODS AND RESULTS: BP was measured in 920 healthy children and adolescents (African-American, Asian, and Caucasian, ages 5 to 18 years). Fat distribution was determined by skinfold thickness and dual-energy X ray absorptiometry (DXA). Pubertal status was assessed by the criteria of Tanner. Regression analysis was used to explore the association between BP and fat distribution. Significant positive relationships between systolic and diastolic BP and trunk fat adjusted for total fat were seen in boys at all pubertal stages in all 3 races by both DXA and skinfold measurements. In girls, trunk fat was not a significant predictor of BP. CONCLUSIONS: Our results demonstrate a sex difference in the relationship between BP and trunk fat in that a significant positive relationship was present in boys only. These findings, based on 2 independent measures of fat distribution, may help identify the specific features of individuals at risk, allow earlier intervention, and suggest sex-specific determinants for BP. PMID- 11877362 TI - Stenting of the arterial duct and banding of the pulmonary arteries: basis for combined Norwood stage I and II repair in hypoplastic left heart. AB - BACKGROUND: Outcome of patients with hypoplastic left heart (HLH) is mainly influenced by the successful first-step palliation according to the Norwood procedure. An alternative approach is heart transplantation (HTX). The feasibility of ductal stenting in newborns with duct-dependent systemic blood flow and bilateral pulmonary artery banding has been reported. But it remains to be elucidated whether this approach allows a new strategy for patients with HLH. METHODS AND RESULTS: In patients with various forms of HLH (n=11) and prostaglandin E-1 administration, ductal stenting was performed with balloon expandable Jo stents or Saxx stents. Bilateral pulmonary artery banding was surgically accomplished 1 to 3 days after the transcatheter procedure. Unrestricted blood flow through the interatrial septum was secured by balloon dilatation atrial septotomy, as required. Interventional procedures were performed with no mortality. Stent and ductal patency were achieved for up to 331 days. Two patients underwent HTX, and 8 patients had a palliative 1-stage procedure with reconstruction of the aortic arch and bidirectional cavopulmonary connection at the age of 3.5 to 6 months. There were 2 deaths. One patient with preoperative right heart failure died after the reconstructive surgery, and 1 patient died 4 months after ductal stenting and bilateral banding awaiting HTX. CONCLUSIONS: The present study is the first clinical trial showing that stenting the duct followed by bilateral pulmonary artery banding in newborns with HLH allows the combination of neoaortic reconstruction, which is part of first-stage palliation of HLH, with the establishment of a bidirectional cavopulmonary connection. Additionally, it allows the chance for HTX after extended waiting periods. PMID- 11877363 TI - Vascular gene transfer of phosphomimetic endothelial nitric oxide synthase (S1177D) using ultrasound-enhanced destruction of plasmid-loaded microbubbles improves vasoreactivity. AB - BACKGROUND: Local gene therapy has enormous potential for the treatment of vascular disease. We determined whether diagnostic ultrasound-mediated destruction of plasmid-loaded albumin microbubbles is a feasible and efficient technique for local vascular gene delivery. For gene transfer, we used a phosphomimetic, active endothelial nitric oxide synthase (eNOS) construct in which Ser1177 was replaced by aspartic acid (S1177D) and exhibits a 2-fold higher basal activity than the wild-type enzyme. METHODS AND RESULTS: Gas-filled microbubbles (3.0 +/- 1.2 microm) were created by sonication of 5% human albumin in the presence of plasmid DNA encoding for LacZ or eNOS S1177D. Porcine coronary arteries were perfused with DNA-loaded albumin microbubbles in vitro, exposed to diagnostic ultrasound (5 seconds), and incubated for a further 24 hours. Detection of the beta-galactosidase in LacZ-transfected vessels revealed a predominant staining of endothelial cells without any functional impairment of vasoreactivity. Western blotting demonstrated the expression of the eNOS S1177D construct in extracts from the transfected segments. Vascular responsiveness was tested with prostaglandin F(2alpha) and the NOS inhibitor N(omega)nitro-L arginine. Compared with segments treated with the expression plasmid alone, the contractile response to prostaglandin F(2alpha) was impaired in segments transfected with eNOS S1177D, whereas the contractile response to the administration of N(omega)nitro-L-arginine was markedly enhanced. CONCLUSIONS: Ultrasound-mediated destruction of eNOS S1177D DNA-loaded albumin microbubbles is a feasible and efficient method for vascular gene transfection. Transfection resulted in significant protein expression and enhanced NO-mediated relaxation of bradykinin-stimulated porcine coronary arteries. PMID- 11877364 TI - Vascular endothelial growth factor is necessary in the development of arteriosclerosis by recruiting/activating monocytes in a rat model of long-term inhibition of nitric oxide synthesis. AB - BACKGROUND: It remains unclear whether vascular endothelial growth factor (VEGF) is a proarteriosclerotic or an antiarteriosclerotic factor. We recently reported that long-term inhibition of nitric oxide by administering Nomega-nitro-L arginine methyl ester (L-NAME) induces coronary vascular inflammation and arteriosclerosis. METHODS AND RESULTS: We used this animal model to investigate the role of VEGF in arteriosclerosis. We blocked VEGF activity in vivo by transfecting with plasmid DNA encoding the murine soluble FLT-1 (sFLT-1) gene into thigh muscle. Soluble FLT-1 can suppress VEGF activity both by sequestering VEGF and by functioning as a dominant-negative inhibitor of VEGF receptors. We observed vascular inflammation associated with increased VEGF expression within 3 days of L-NAME administration, which was prevented by pretreatment with ACE inhibitor, angiotensin II receptor antagonist, or neutralizing monocyte chemoattractant protein-1 antibody. The sFLT-1 gene transfer attenuated the early vascular inflammation and prevented late arteriosclerosis. The sFLT-1 gene transfer also inhibited increased expression of monocyte chemoattractant protein 1 and transforming growth factor-beta, indicating creation of a positive feedback loop to cause arteriosclerosis. CONCLUSIONS: VEGF is necessary in the development of arteriosclerosis by mediating monocyte recruitment and activation in this model. PMID- 11877365 TI - Evidence for synergy between alpha(2)-adrenergic and nonadrenergic mechanisms in central blood pressure regulation. AB - BACKGROUND: Both alpha(2)-adrenergic and non--alpha(2)-adrenergic mechanisms seem to be involved in the hypotensive effect of imidazoline-like drugs. This study aimed at investigating how these 2 mechanisms work together to modify blood pressure (BP). METHODS AND RESULTS: LNP 509, which appeared in this study to be devoid of alpha(2A)-adrenergic activity, was administered to anesthetized rabbits and wild-type (WT) mice into the cisterna magna and into the fourth ventricle, respectively. Mean arterial pressure decreased by a maximum of 46 +/- 4% and 16 +/- 2%, respectively. In D79N mice, which lack functional alpha(2A)-adrenergic receptors, LNP 509 also reduced mean arterial pressure by 17 +/- 2%. The hypotension induced by LNP 509 (100 microg/kg intracisternally) was prevented by S23757 (1 mg/kg intracisternally), an antagonist highly selective for I(1) imidazoline binding sites (I(1)BS). A synergy between LNP 509 and the alpha(2) adrenergic agonist alpha-methylnoradrenaline (alpha-MNA) was observed in rabbits (cisterna magna injection) and in WT mice (fourth ventricle injection) but not, as expected, in D79N mice. Similar to LNP 509 alone, rilmenidine (fourth ventricle injection), which binds both to alpha(2)-adrenergic receptors and to I(1)BS, decreased BP in D79N mice. In WT animals, rilmenidine had a significantly greater effect. Microinjections performed in rabbits showed that the synergism occurred at least in part in the nucleus reticularis lateralis of the brain stem. CONCLUSIONS: These results demonstrate that a central imidazoline-sensitive, but non--alpha(2)-adrenergic, mechanism can modify BP by itself. This mechanism, which may involve I(1)BS, interacts synergistically with an alpha(2)-adrenergic mechanism to decrease BP. PMID- 11877366 TI - Endogenous ligand of alpha(1) sodium pump, marinobufagenin, is a novel mediator of sodium chloride--dependent hypertension. AB - BACKGROUND: Digitalis-like sodium pump ligands (SPLs) effect natriuresis via inhibition of renal tubular Na(+),K(+)-ATPase but may induce vasoconstriction. The present study investigated the potential roles of 2 putative endogenous SPLs, an ouabain-like compound (OLC) and an alpha(1) Na(+),K(+)-ATPase inhibitor, marinobufagenin (MBG), in regulating natriuresis and blood pressure (BP) responses to sustained and acute NaCl loading in Dahl salt-sensitive rats (DS). METHODS AND RESULTS: During 4 weeks of an 8% NaCl diet, DS exhibited a progressive increase in MBG renal excretion (66 +/-13 pmol/24 hours at week 4 versus 11 +/- 1 pmol/24 hours at baseline, n=48), which paralleled an increase in systolic BP (174 +/- 10 mm Hg at week 4 versus 110 +/- 2 mm Hg at baseline). By contrast, OLC excretion peaked at week 1 and returned to baseline levels. Administration of an anti-MBG, but not anti-ouabain antibody, to DS after 3 weeks of a high NaCl diet lowered BP (139 +/- 7 versus 175 +/- 5 mm Hg, P<0.001, n=5). Acute NaCl loading (2 hours) of DS (n=5) increased MBG and OLC excretion and natriuresis. Pretreatment of acutely NaCl-loaded DS with an anti-MBG antibody (n=5) reduced the excretion of sodium and MBG but not that of OLC. An anti ouabain antibody (n=5) reduced sodium excretion and both OLC and MBG. CONCLUSIONS: An initial transient stimulation of OLC induced by NaCl loading of DS precedes an MBG response. A sustained increase in MBG production in DS contributes to the chronic BP elevation induced by a sustained high NaCl intake. PMID- 11877367 TI - Electrophysiological mechanism of enhanced susceptibility of hypertrophied heart to acquired torsade de pointes arrhythmias: tridimensional mapping of activation and recovery patterns. AB - BACKGROUND: Cardiac hypertrophy is associated with increased incidence of sudden death and susceptibility to proarrhythmic effects of antiarrhythmic agents. However, the in vivo electrophysiologic mechanism of the arrhythmias has not been investigated in detail. METHODS AND RESULTS: Dose-dependent susceptibility to Torsade de Pointes (TdP) by class III drug dofetilide, 3, 10, and 30 microg/kg, was compared in 6 control dogs (C) and in 5 dogs 6 to 8 weeks after induction of complete atrioventricular block (AVB) that resulted in ventricular hypertrophy (H). Tridimensional ventricular activation and repolarization (R) patterns were simultaneously analyzed from unipolar extracellular electrograms, and local R was measured from activation recovery intervals. Both R and transmural dispersion of R (TDR) were significantly greater in dogs with H compared with C. Dofetilide resulted in cycle length--dependent and dose-dependent prolongation of R, which was more marked in left ventricular endocardium/midmyocardium compared with epicardium, resulting in significant increase of TDR. These changes were more accentuated in dogs with H compared with C. All 5 dogs with H developed TdP at a dose of 3 to 10 microg/kg, whereas only 1 of 6 C dogs developed TdP at 30 microg/kg. TdP was initiated by subendocardial focal activity that infringed on TDR, resulting in functional conduction block and reentrant excitation. CONCLUSIONS: Enhanced susceptibility of hypertrophied heart to class III drugs is attributable to baseline increased TDR and greater dose-related accentuation of TDR compared with nonhypertrophied heart. This provides the electrophysiologic substrate for drug-induced TdP. PMID- 11877368 TI - Inflammation and atherosclerosis. AB - Atherosclerosis, formerly considered a bland lipid storage disease, actually involves an ongoing inflammatory response. Recent advances in basic science have established a fundamental role for inflammation in mediating all stages of this disease from initiation through progression and, ultimately, the thrombotic complications of atherosclerosis. These new findings provide important links between risk factors and the mechanisms of atherogenesis. Clinical studies have shown that this emerging biology of inflammation in atherosclerosis applies directly to human patients. Elevation in markers of inflammation predicts outcomes of patients with acute coronary syndromes, independently of myocardial damage. In addition, low-grade chronic inflammation, as indicated by levels of the inflammatory marker C-reactive protein, prospectively defines risk of atherosclerotic complications, thus adding to prognostic information provided by traditional risk factors. Moreover, certain treatments that reduce coronary risk also limit inflammation. In the case of lipid lowering with statins, this anti inflammatory effect does not appear to correlate with reduction in low-density lipoprotein levels. These new insights into inflammation in atherosclerosis not only increase our understanding of this disease, but also have practical clinical applications in risk stratification and targeting of therapy for this scourge of growing worldwide importance. PMID- 11877369 TI - Positron emission tomography predicted recovery of complete A-V nodal dysfunction in a patient with cardiac sarcoidosis. PMID- 11877370 TI - Automated external defibrillators in health/fitness facilities: supplement to the AHA/ACSM Recommendations for Cardiovascular Screening, Staffing, and Emergency Policies at Health/Fitness Facilities. PMID- 11877371 TI - Images in cardiovascular medicine. Noncompaction of the ventricular myocardium. PMID- 11877372 TI - Predicting stroke after myocardial infarction in the elderly. PMID- 11877373 TI - Nuclear lamins: building blocks of nuclear architecture. PMID- 11877374 TI - Wnt signaling plays an essential role in neuronal specification of the dorsal spinal cord. AB - In the developing spinal cord, signals from the roof plate are required for the development of three classes of dorsal interneuron: D1, D2, and D3, listed from dorsal to ventral. Here, we demonstrate that absence of Wnt1 and Wnt3a, normally expressed in the roof plate, leads to diminished development of D1 and D2 neurons and a compensatory increase in D3 neuron populations. This occurs without significantly altered expression of BMP and related genes in the roof plate. Moreover, Wnt3a protein induces expression of D1 and D2 markers in the isolated medial region of the chick neural plate, and Noggin does not interfere with this induction. Thus, Wnt signaling plays a critical role in the specification of cell types for dorsal interneurons. PMID- 11877375 TI - Arabidopsis COP10 is a ubiquitin-conjugating enzyme variant that acts together with COP1 and the COP9 signalosome in repressing photomorphogenesis. AB - A group of evolutionarily conserved pleiotropic COP/DET/FUS proteins was initially defined by their ability to repress photomorphogenesis in Arabidopsis. It was proposed that this regulation be mediated by targeting degradation of key cellular regulators that promote photomorphogenesis. Among them, COP1 and the COP9 signalosome have been hypothesized to fulfill the roles as an ubiquitin ligase (E3) and an essential E3 modulator. Here we report that COP10 encodes a protein similar to ubiquitin-conjugating enzyme (E2) variant proteins (UEV). COP10 is part of a nuclear protein complex and capable of directly interacting with both COP1 and the COP9 signalosome. Our data indicates that COP10 defines a possible E2 activity, thus validating the working hypothesis that the pleiotropic COP/DET/FUS group of proteins defined a protein ubiquitination pathway. PMID- 11877376 TI - Involvement of the cohesin protein, Smc1, in Atm-dependent and independent responses to DNA damage. AB - Structural maintenance of chromosomes (SMC) proteins play important roles in sister chromatid cohesion, chromosome condensation, sex-chromosome dosage compensation, and DNA recombination and repair. Protein complexes containing heterodimers of the Smc1 and Smc3 proteins have been implicated specifically in both sister chromatid cohesion and DNA recombination. Here, we show that the protein kinase, Atm, which belongs to a family of phosphatidylinositol 3-kinases that regulate cell cycle checkpoints and DNA recombination and repair, phosphorylates Smc1 protein after ionizing irradiation. Atm phosphorylates Smc1 on serines 957 and 966 in vitro and in vivo, and expression of an Smc1 protein mutated at these phosphorylation sites abrogates the ionizing irradiation-induced S phase cell cycle checkpoint. Optimal phosphorylation of these sites in Smc1 after ionizing irradiation also requires the presence of the Atm substrates Nbs1 and Brca1. These same sites in Smc1 are phosphorylated after treatment with UV irradiation or hydroxyurea in an Atm-independent manner, thus demonstrating that another kinase must be involved in responses to these cellular stresses. Yeast containing hypomorphic mutations in SMC1 and human cells overexpressing Smc1 mutated at both of these phosphorylation sites exhibit decreased survival following ionizing irradiation. These results demonstrate that Smc1 participates in cellular responses to DNA damage and link Smc1 to the Atm signal transduction pathway. PMID- 11877377 TI - SMC1 is a downstream effector in the ATM/NBS1 branch of the human S-phase checkpoint. AB - Structural maintenance of chromosomes (SMC) proteins (SMC1, SMC3) are evolutionarily conserved chromosomal proteins that are components of the cohesin complex, necessary for sister chromatid cohesion. These proteins may also function in DNA repair. Here we report that SMC1 is a component of the DNA damage response network that functions as an effector in the ATM/NBS1-dependent S-phase checkpoint pathway. SMC1 associates with BRCA1 and is phosphorylated in response to IR in an ATM- and NBS1-dependent manner. Using mass spectrometry, we established that ATM phosphorylates S957 and S966 of SMC1 in vivo. Phosphorylation of S957 and/or S966 of SMC1 is required for activation of the S phase checkpoint in response to IR. We also discovered that the phosphorylation of NBS1 by ATM is required for the phosphorylation of SMC1, establishing the role of NBS1 as an adaptor in the ATM/NBS1/SMC1 pathway. The ATM/CHK2/CDC25A pathway is also involved in the S-phase checkpoint activation, but this pathway is intact in NBS cells. Our results indicate that the ATM/NBS1/SMC1 pathway is a separate branch of the S-phase checkpoint pathway, distinct from the ATM/CHK2/CDC25A branch. Therefore, this work establishes the ATM/NBS1/SMC1 branch, and provides a molecular basis for the S-phase checkpoint defect in NBS cells. PMID- 11877378 TI - Structure of the 53BP1 BRCT region bound to p53 and its comparison to the Brca1 BRCT structure. AB - Brca1 C-terminal (BRCT) domains are a common protein-protein interaction motif in proteins involved in the DNA damage response and DNA repair. The DNA-damage response protein 53BP1 has two BRCT domains that bind to the DNA-binding domain of p53. The 53BP1 tandem-BRCT region is homologous to the tandem-BRCT region of Brca1, which is involved in double-strand break repair and homologous recombination and which binds BACH1, a member of the DEAH helicase family. Here we report the structures of a human 53BP1-p53 complex and of the rat Brca1 BRCT repeats. The 53BP1-p53 structure shows that the two BRCT repeats are arranged tandemly and pack extensively through an interface that also involves the inter repeat linker. The first BRCT repeat and the linker together bind p53 on a region that overlaps with the DNA-binding surface of p53 and involves p53 residues that are mutated in cancer and are important for DNA binding. Comparison with the structure of the tandem-BRCT region of Brca1 shows a remarkable conservation of the repeat arrangement and of the inter-BRCT repeat interface. Analysis of human BRCA1 tumor-derived mutations and conservation identifies a potential protein binding site that we show through mutagenesis is involved in BACH1 binding. The BACH1-binding region of Brca1 consists of a unique insertion in the first BRCT repeat and the inter-repeat linker and is analogous to the region of 53BP1 that binds p53. PMID- 11877379 TI - Stability of a PKCI-1-related mRNA is controlled by the splicing factor ASF/SF2: a novel function for SR proteins. AB - Pre-mRNA splicing is a widely used regulatory mechanism for controlling gene expression, and a family of conserved proteins, SR proteins, participate in both constitutive and alternative splicing. Here we describe a novel function for the SR protein ASF/SF2. We used an embryonic chicken cDNA library to screen for differential mRNA expression in the chicken B-cell line DT40-ASF, expressing or not expressing ASF/SF2. Remarkably, out of 3 x 10(6) clones screened, only one, isolated several times independently, showed ASF/SF2-related differential expression. The isolated cDNA, referred to here as PKCI-r (for PKCI-related), is closely related to the protein kinase C interacting protein (PKCI-1) gene. Transcript levels were increased approximately sixfold in ASF/SF2-depleted cells compared with cells expressing ASF/SF2, indicating a negative role for the SR protein. Strikingly, inhibition of ASF/SF2 expression had no significant effect on PKCI-r splicing, or transcription, but markedly increased the half-life of PKCI-r mRNA (6.6-fold). Similarly, increased mRNA stability was also observed upon expression of exogenous PKCI-r mRNA in cells depleted of ASF/SF2. ASF/SF2 bound to a discrete region containing a purine-rich sequence in the 3' UTR of the PKCI-r transcript, and deletion of this region eliminated ASF/SF2-mediated regulation of transcript stability. Together these data indicate a novel, direct effect of ASF/SF2 on PKCI-r mRNA stability. Therefore, ASF/SF2, and perhaps other SR proteins, affects gene expression in vertebrate cells through regulation of mRNA stability as well as splicing. PMID- 11877380 TI - Calcineurin-dependent regulation of Crz1p nuclear export requires Msn5p and a conserved calcineurin docking site. AB - Calcineurin, a conserved Ca(2+)/calmodulin-regulated protein phosphatase, plays a crucial role in Ca(2+) signaling in a wide variety of cell types. In Saccharomyces cerevisiae, calcineurin positively regulates transcription in response to stress by dephosphorylating the transcription factor Crz1p/Tcn1p. Dephosphorylation promotes Crz1p nuclear localization in part by increasing the efficiency of its nuclear import. In this work, we show that calcineurin dependent dephosphorylation of Crz1p also down-regulates its nuclear export. Using a genetic approach, we identify Msn5p as the exportin for Crz1p. In addition, we define the Crz1p nuclear export signal (NES) and show that it interacts with Msn5p in a phosphorylation-dependent manner. This indicates that calcineurin regulates Crz1p nuclear export by dephosphorylating and inactivating its NES. Finally, we define a motif in Crz1p, PIISIQ, similar to the PxIxIT docking site for calcineurin on the mammalian transcription factor NFAT, that mediates the in vivo interaction between calcineurin and Crz1p and is required for calcineurin-dependent regulation of Crz1p nuclear export and activity. Therefore, in yeast as in mammals, a docking site is required to target calcineurin to its substrate such that it can dephosphorylate it efficiently. PMID- 11877381 TI - The GEX-2 and GEX-3 proteins are required for tissue morphogenesis and cell migrations in C. elegans. AB - During body morphogenesis precisely coordinated cell movements and cell shape changes organize the newly differentiated cells of an embryo into functional tissues. Here we describe two genes, gex-2 and gex-3, whose activities are necessary for initial steps of body morphogenesis in Caenorhabditis elegans. In the absence of gex-2 and gex-3 activities, cells differentiate properly but fail to become organized. The external hypodermal cells fail to spread over and enclose the embryo and instead cluster on the dorsal side. Postembryonically gex 3 activity is required for egg laying and for proper morphogenesis of the gonad. GEX-2 and GEX-3 proteins colocalize to cell boundaries and appear to directly interact. GEX-2 and GEX-3 are highly conserved, with vertebrate homologs implicated in binding the small GTPase Rac and a GEX-3 Drosophila homolog, HEM2/NAP1/KETTE, that interacts genetically with Rac pathway mutants. Our findings suggest that GEX-2 and GEX-3 may function at cell boundaries to regulate cell migrations and cell shape changes required for proper morphogenesis and development. PMID- 11877382 TI - Distinct requirements for TrkB and TrkC signaling in target innervation by sensory neurons. AB - Signaling by brain-derived neurotrophic factor (BDNF) via the TrkB receptor, or by neurotrophin-3 (NT3) through the TrkC receptor support distinct populations of sensory neurons. The intracellular signaling pathways activated by Trk (tyrosine kinase) receptors, which in vivo promote neuronal survival and target innervation, are not well understood. Using mice with TrkB or TrkC receptors lacking the docking site for Shc adaptors (trkB(shc/shc) and trkC(shc/shc) mice), we show that TrkB and TrkC promote survival of sensory neurons mainly through Shc site-independent pathways, suggesting that these receptors use similar pathways to prevent apoptosis. In contrast, the regulation of target innervation appears different: in trkB(shc/shc) mice neurons lose target innervation, whereas in trkC(shc/shc) mice the surviving TrkC-dependent neurons maintain target innervation and function. Biochemical analysis indicates that phosphorylation at the Shc site positively regulates autophosphorylation of TrkB, but not of TrkC. Our findings show that although TrkB and TrkC signals mediating survival are largely similar, TrkB and TrkC signals required for maintenance of target innervation in vivo are regulated by distinct mechanisms. PMID- 11877384 TI - The antidiabetic agent LG100754 sensitizes cells to low concentrations of peroxisome proliferator-activated receptor gamma ligands. AB - Insulin resistance and non-insulin-dependent diabetes mellitus are major causes of morbidity and mortality in industrialized nations. Despite the alarming rise in the prevalence of this disorder, the initial molecular events that promote insulin resistance remain unclear. The data presented here demonstrate that LG100754, an antidiabetic RXR ligand, defines a novel type of nuclear receptor agonist. Surprisingly, LG100754 has minimal intrinsic transcriptional activity, instead it enhances the potency of proliferator-activated receptor (PPAR) gamma retinoid X receptor heterodimers for PPARgamma ligands. The ability of LG100754 to both increase PPARgamma sensitivity and relieve insulin resistance implies that a deficiency in endogenous PPARgamma ligands may represent an early step in the development of insulin resistance. PMID- 11877383 TI - Gibberellin regulates Arabidopsis seed germination via RGL2, a GAI/RGA-like gene whose expression is up-regulated following imbibition. AB - The germination of Arabidopsis seeds is promoted by gibberellin (GA). Arabidopsis GAI, and RGA are genes encoding key GA signal-transduction components (GAI and RGA) that mediate GA regulation of stem elongation. The Arabidopsis genome contains two further genes, RGL1 and RGL2, that encode proteins (RGL1 and RGL2) that are closely related to GAI and RGA. Here, we show that RGL2 regulates seed germination in response to GA, and that RGL1, GAI, and RGA do not. In addition, we show that RGL2 transcript levels rise rapidly following seed imbibition, and then decline rapidly as germination proceeds. In situ GUS staining revealed that RGL2 expression in imbibed seeds is restricted to elongating regions of pre emergent and recently emerged radicles. These observations indicate that RGL2 is a negative regulator of GA responses that acts specifically to control seed germination rather than stem elongation. Furthermore, as RGL2 expression is imbibition inducible, RGL2 may function as an integrator of environmental and endogenous cues to control seed germination. PMID- 11877385 TI - FRS2 PTB domain conformation regulates interactions with divergent neurotrophic receptors. AB - Membrane-anchored adaptor proteins FRS2alpha/beta (also known as SNT-1/2) mediate signaling of fibroblast growth factor receptors (FGFRs) and neurotrophin receptors (TRKs) through their N-terminal phosphotyrosine binding (PTB) domains. The FRS2 PTB domain recognizes tyrosine-phosphorylated TRKs at an NPXpY (where pY is phosphotyrosine) motif, whereas its constitutive association with FGFR involves a receptor juxtamembrane region lacking Tyr and Asn residues. Here we show by isothermal titration calorimetry that the FRS2alpha PTB domain binding to peptides derived from TRKs or FGFR is thermodynamically different. TRK binding is largely enthalpy-driven, whereas the FGFR interaction is governed by a favorable entropic contribution to the free energy of binding. Furthermore, our NMR spectral analysis suggests that disruption of an unstructured region C-terminal to the PTB domain alters local conformation and dynamics of the residues at the ligand-binding site, and that structural disruption of the beta8-strand directly weakens the PTB domain association with the FGFR ligand. Together, our new findings support a molecular mechanism by which conformational dynamics of the FRS2alpha PTB domain dictates its association with either fibroblast growth factor or neurotrophin receptors in neuronal development. PMID- 11877386 TI - Structural determinants of BRCA1 translational regulation. AB - The BRCA1 gene is involved in sporadic breast and ovarian cancer mainly through reduced expression. BRCA1 mRNAs containing different leader sequences show different patterns of expression. In a normal mammary gland mRNA with a shorter leader sequence, 5'-UTRa is expressed only, whereas in breast cancer tissue mRNA with a longer leader, 5'-UTRb is expressed also. We show that the translation efficiency of transcripts containing 5'-UTRb is 10 times lower than those containing 5'-UTRa. The structures of 5'-UTRa and 5'-UTRb were determined by chemical and enzymatic probing aided by a new method developed for monitoring the number of co-existing stable conformers. Specific factors responsible for reduced translation of mRNA containing 5'-UTRb were determined using a variety of transcripts with mutations in the leader sequence. These factors include a stable secondary structure formed by truncated Alu element and upstream AUG codons. The novel mechanism by which BRCA1 may be involved in sporadic breast and ovarian cancer is proposed. It is based on the expression patterns of BRCA1 mRNAs and differences in their translatability. According to this mechanism the deregulation of the BRCA1 transcription in cancer, resulting in a higher proportion of translationally inhibited transcripts containing 5'-UTRb, contributes to the decrease in the BRCA1 protein observed in sporadic breast and ovarian cancers. PMID- 11877387 TI - UDP-glucuronate decarboxylase, a key enzyme in proteoglycan synthesis: cloning, characterization, and localization. AB - UDP-glucuronate decarboxylase (UGD) catalyzes the formation of UDP-xylose from UDP-glucuronate. UDP-xylose is then used to initiate glycosaminoglycan biosynthesis on the core protein of proteoglycans. In a yeast two-hybrid screen with the protein kinase Akt (protein kinase B), we detected interactions with a novel sequence, which we cloned and expressed. The expressed protein displayed UGD activity but did not display the activities of homologous nucleotide sugar epimerases or dehydratases. We did not detect phosphorylation of UGD by Akt nor did we detect any influence of Akt on UGD activity. Effects of UGD on Akt kinase activity were also absent. Northern blot and Western blot analyses revealed the presence of UGD in multiple tissues and brain regions. Subcellular studies and histochemistry localized UGD protein to the perinuclear Golgi where xylosylation of proteoglycan core proteins is known to occur. PMID- 11877388 TI - Hyperoxia-induced apoptosis does not require mitochondrial reactive oxygen species and is regulated by Bcl-2 proteins. AB - Exposure of animals to hyperoxia results in lung injury that is characterized by apoptosis and necrosis of the alveolar epithelium and endothelium. The mechanism by which hyperoxia results in cell death, however, remains unclear. We sought to test the hypothesis that exposure to hyperoxia causes mitochondria-dependent apoptosis that requires the generation of reactive oxygen species from mitochondrial electron transport. Rat1a cells exposed to hyperoxia underwent apoptosis characterized by the release of cytochrome c, activation of caspase-9, and nuclear fragmentation that was prevented by the overexpression of Bcl-X(L.) Murine embryonic fibroblasts from bax(-/-) bak(-/-) mice were resistant to hyperoxia-induced cell death. The administration of the antioxidants manganese (III) tetrakis (4-benzoic acid) porphyrin, ebselen, and N-acetylcysteine failed to prevent cell death following exposure to hyperoxia. Human fibrosarcoma cells (HT1080) lacking mitochondrial DNA (rho(0) cells) that failed to generate reactive oxygen species during exposure to hyperoxia were not protected against cell death following exposure to hyperoxia. We conclude that exposure to hyperoxia results in apoptosis that requires Bax or Bak and can be prevented by the overexpression of Bcl-X(L). The mitochondrial generation of reactive oxygen species is not required for cell death following exposure to hyperoxia. PMID- 11877389 TI - Glucocorticoids inhibit cell cycle progression in differentiating osteoblasts via glycogen synthase kinase-3beta. AB - Differentiating osteoblasts in culture undergo a commitment stage, during which cobblestone-like cells grow to high density past confluency. In contrast to earlier proliferative stages, the cell cycle during this commitment stage is inhibited by glucocorticoids (GC). Chronic GC treatment also impedes mineral deposition if steroid administration commences early enough during commitment. This study defines a role for glycogen synthase kinase-3beta (GSK3beta) and its target, c-Myc, in the GC-sensitive osteoblast persistent cell cycle. c-Myc levels decreased as cells reached confluence, but then increased during growth to high density. GC administration at this stage resulted in down-regulation of c-Myc. This was accompanied by GC-mediated attenuation of GSK3beta Ser(9) inhibitory phosphorylation and increased GSK3beta kinase activity. Down-regulation of c-Myc was attributable to enhanced Thr(58) phosphorylation, leading to accelerated degradation. In contrast, GC did not inhibit the c-Myc synthesis rate or the level of beta-catenin, a transcriptional coactivator of c-myc. The attenuated cell cycle and the reduced c-Myc level were returned to control levels by specific inhibition of GSK3beta using lithium chloride. These results suggest that tonal GSK3beta repression at the cobblestone stage of osteoblast differentiation permits osteoblast growth to high density. GC interfere with this growth-permissive axis by GSK3beta activation, resulting in c-Myc down-regulation and impediment of the G(1)/S cell cycle transition. PMID- 11877390 TI - ANGPTL3 stimulates endothelial cell adhesion and migration via integrin alpha vbeta 3 and induces blood vessel formation in vivo. AB - The angiopoietin family of secreted factors is functionally defined by the C terminal fibrinogen (FBN)-like domain, which mediates binding to the Tie2 receptor and thereby facilitates a cascade of events ultimately regulating blood vessel formation. By screening expressed sequence tag data bases for homologies to a consensus FBN-like motive, we have identified ANGPTL3, a liver-specific, secreted factor consisting of an N-terminal coiled-coil domain and the C-terminal FBN-like domain. Co-immunoprecipitation experiments, however, failed to detect binding of ANGPTL3 to the Tie2 receptor. A molecular model of the FBN-like domain of ANGPTL3 was generated and predicted potential binding to integrins. This hypothesis was experimentally confirmed by the finding that recombinant ANGPTL3 bound to alpha(v)beta(3) and induced integrin alpha(v)beta(3)-dependent haptotactic endothelial cell adhesion and migration and stimulated signal transduction pathways characteristic for integrin activation, including phosphorylation of Akt, mitogen-activated protein kinase, and focal adhesion kinase. When tested in the rat corneal assay, ANGPTL3 strongly induced angiogenesis with comparable magnitude as observed for vascular endothelial growth factor-A. Moreover, the C-terminal FBN-like domain alone was sufficient to induce endothelial cell adhesion and in vivo angiogenesis. Taken together, our data demonstrate that ANGPTL3 is the first member of the angiopoietin-like family of secreted factors binding to integrin alpha(v)beta(3) and suggest a possible role in the regulation of angiogenesis. PMID- 11877392 TI - Protein kinase C and calcium/calmodulin-activated protein kinase II (CaMK II) suppress nicotinic acetylcholine receptor gene expression in mammalian muscle. A specific role for CaMK II in activity-dependent gene expression. AB - Nicotinic acetylcholine receptor (nAChR) gene expression is regulated by both muscle activity and increased intracellular calcium. This regulation is an important developmental event that rids receptors from the extrajunctional region of the developing muscle fiber. In avian muscle, it has been proposed that muscle activity suppresses nAChR gene expression via calcium-activated protein kinase C (PKC)-dependent phosphorylation of the myogenic transcription factor, myogenin. Here, we examined the role that PKC and other kinases play in mediating calcium- and activity-dependent suppression of nAChR genes in rat primary myotubes. We found that although activated PKC could regulate nAChR promoter activity and transiently suppressed both nAChR and myogenin gene expression, it did not appear to be required for calcium- or activity-dependent control of nAChR gene expression in mammalian muscle. Neither depletion of PKC from myotubes nor specific pharmacological inhibition of PKC blocked the suppression of nAChR gene expression produced by calcium or muscle depolarization. In contrast, we provide evidence that calcium/calmodulin-activated protein kinase II participates in mediating the effects of muscle depolarization on nAChR and myogenin gene expression. PMID- 11877391 TI - Phosphatidylinositol 4-phosphate 5-kinase is essential for ROCK-mediated neurite remodeling. AB - Phosphatidylinositol 4-phosphate 5-kinase (PIP-5kin) regulates actin cytoskeletal reorganization through its product phosphatidylinositol 4,5-bisphosphate. In the present study we demonstrate that PIP-5kin is essential for neurite remodeling, which is regulated by actin cytoskeletal reorganization in neuroblastoma N1E-115 cells. Overexpression of wild-type mouse PIP-5kin-alpha inhibits the neurite formation that is normally stimulated by serum depletion, whereas a lipid kinase defective mutant of PIP-5kin-alpha, D266A, triggers neurite extension even in the presence of serum and blocks lysophosphatidic acid-induced neurite retraction. These results phenocopy those previously reported for the small GTPase RhoA and its effector p160 Rho-associated coiled coil-forming protein kinase (ROCK). However, the ROCK-specific inhibitor Y-27632 failed to block the inhibition by PIP-5kin-alpha of neurite extension, whereas D266A did block the neurite retraction induced by overexpression of ROCK. These results, taken together, suggest that PIP-5kin-alpha functions as a downstream effector for RhoA/ROCK to couple lysophosphatidic acid signaling to neurite retraction presumably through its product phosphatidylinositol 4,5-bisphosphate. PMID- 11877393 TI - Altered amelogenin self-assembly based on mutations observed in human X-linked amelogenesis imperfecta (AIH1). AB - A hallmark of biological systems is a reliance on protein assemblies to perform complex functions. We have focused attention on mammalian enamel formation because it relies on a self-assembling protein complex to direct mineral habit. The principle protein of enamel is amelogenin, a 180-amino acid hydrophobic protein that self-assembles to form nanospheres. We have used independent technical methods, consisting of the yeast two-hybrid (Y2H) assay and surface plasmon resonance (SPR), to demonstrate the importance of amelogenin self assembly domains. In addition, we have analyzed mutations in amelogenin observed in patients with amelogenesis imperfecta who demonstrate defects in enamel formation. Assessments of self-assembly of these mutant amelogenins by either SPR or Y2H assay yield concordant data. These data support the conclusion that the amelogenin amino-terminal self-assembly domain is essential to the creation of an enamel extracellular organic matrix capable of directing mineral formation. It also suggests that a pathway through which point mutations in the amelogenin protein can adversely impact on the formation of the enamel organ is by disturbing self-assembly of the organic matrix. These data support the utilization of the Y2H assay to search for protein interactions among extracellular matrix proteins that contribute to biomineralization and provide functional information on protein-protein and protein-mineral interactions. PMID- 11877394 TI - Insulin receptor substrate 1 translocation to the nucleus by the human JC virus T antigen. AB - Insulin receptor substrate 1 (IRS-1) is the major signaling molecule for the insulin and insulin-like growth factor I receptors, which transduces both metabolic and growth-promoting signals, and has transforming properties when overexpressed in the cells. Here we show that IRS-1 is translocated to the nucleus in the presence of the early viral protein-T-antigen of the human polyomavirus JC. Nuclear IRS-1 was detected in T-antigen-positive cell lines and in T-antigen-positive biopsies from patients diagnosed with medulloblastoma. The IRS-1 domain responsible for a direct JC virus T-antigen binding was localized within the N-terminal portion of IRS-1 molecule, and the binding was independent from IRS-1 tyrosine phosphorylation and was strongly inhibited by IRS-1 serine phosphorylation. In addition, competition for the IRS-1-T-antigen binding by a dominant negative mutant of IRS-1 inhibited growth and survival of JC virus T antigen-transformed cells in anchorage-independent culture conditions. Based on these findings, we propose a novel role for the IRS-1-T-antigen complex in controlling cellular equilibrium during viral infection. It may involve uncoupling of IRS-1 from its surface receptor and translocation of its function to the nucleus. PMID- 11877395 TI - Insulin-like growth factor I induces MDM2-dependent degradation of p53 via the p38 MAPK pathway in response to DNA damage. AB - In many tissues, the insulin-like growth factor I (IGF-I) receptor (IGF-IR) is known to functionally oppose apoptosis. Recently, we demonstrated a direct role for the IGF-IR in the rescue of DNA-damaged fibroblasts by activating a DNA repair pathway (Heron-Milhavet, L., Karas, M., Goldsmith, C. M., Baum, B. J., and LeRoith, D. (2001) J. Biol. Chem. 276, 18185-18192). p53 is a nuclear transcription factor that can block progression of the cell cycle, modulate DNA repair, and trigger apoptosis. In this work, we tested the effect of IGF-I on the regulation of the p53 signaling cascade. The DNA-damaging agent 4-nitroquinoline 1-oxide was applied to NIH-3T3 cells overexpressing normal IGF-IRs (NWTb3 cells). We showed that after 4-nitroquinoline 1-oxide-induced DNA damage, IGF-I induced exclusion of the p53 protein from the nucleus and led to its degradation in the cytoplasm, whereas p53 mRNA was unaffected. Degradation of the p53 protein was associated with an increase in MDM2, an upstream modulator of the half-life and activity of the p53 protein. p53 degradation was also associated with down regulation of p21. We further showed that the effects of IGF-I on mdm2 transcription and on MDM2/p19 ARF association were mediated by the p38 MAPK pathway. In conclusion, we describe a novel role for IGF-I in the regulation of the MDM2/p53/p21 signaling pathway during DNA damage. PMID- 11877396 TI - Bacteriophage phi 6 RNA-dependent RNA polymerase: molecular details of initiating nucleic acid synthesis without primer. AB - Like most RNA polymerases, the polymerase of double-strand RNA bacteriophage phi6 (phi6pol) is capable of primer-independent initiation. Based on the recently solved phi6pol initiation complex structure, a four-amino acid-long loop (amino acids 630-633) has been suggested to stabilize the first two incoming NTPs through stacking interactions with tyrosine, Tyr(630). A similar loop is also present in the hepatitis C virus polymerase, another enzyme capable of de novo initiation. Here, we use a series of phi6pol mutants to address the role of this element. As predicted, mutants at the Tyr(630) position are inefficient in initiation de novo. Unexpectedly, when the loop is disordered by changing Tyr(630)-Lys(631)-Trp(632) to GSG, phi6pol becomes a primer-dependent enzyme, either extending complementary oligonucleotide or, when the template 3' terminus can adopt a hairpin-like conformation, utilizing a "copy-back" initiation mechanism. In contrast to the wild-type phi6pol, the GSG mutant does not require high GTP concentration for its optimal activity. These findings suggest a general model for the initiation of de novo RNA synthesis. PMID- 11877397 TI - Stimulation of IRF-7 gene expression by tumor necrosis factor alpha: requirement for NFkappa B transcription factor and gene accessibility. AB - Interferon regulatory factor 7 (IRF-7) plays an important role in innate immunity, where, together with IRF-3, it controls the expression of interferon A/B genes as well as chemokine RANTES (regulated on activation normal T cell expressed and secreted). Previously, we characterized human IRF-7 promoter and showed that an interferon-stimulated response element site in the first intron binds interferon-stimulated gene factor 3 (ISGF3) and confers the response to interferon. Here we report the stimulation of IRF-7 expression by 12-O tetradecanoylphorbol-13-acetate (TPA) and tumor necrosis factor alpha (TNFalpha) in human peripheral blood monocytes. Using promoter analysis in combination with electrophoretic mobility shift assays, we have demonstrated that an NFkappaB site located next to the TATA box, binds p50 and p65 heterodimer and is required for the induction of the IRF-7 gene by TPA and TNFalpha. In addition, we report stimulation of IRF-7 gene expression by topoisomerase II (TOPII) inhibitors. We show by chromatin immunoprecipitation assay that treatment with the TOPII inhibitor etoposide induces association of acetylated histone 3 with the promoter of IRF-7 gene, indicating that TOPII-mediated changes in chromatin structure could be responsible for the induction. This suggests that the IRF-7 gene is localized in the condensed area of the chromosome where it is inaccessible to transcription factors that would promote its constitutive expression. PMID- 11877398 TI - Protein kinase C inhibits type VI adenylyl cyclase by phosphorylating the regulatory N domain and two catalytic C1 and C2 domains. AB - We previously showed that phosphorylation of Ser(10) of the N terminus domain of the type VI adenylyl cyclase (ACVI) partly mediated protein kinase C (PKC) induced inhibition of ACVI. We now report that phosphorylation of the other two cytosolic domains (C1 and C2), which form the catalytic core complex of ACVI, also contributes to PKC-mediated inhibition. In vitro phosphorylation by PKC of the recombinant C1a and C2 domains, and of the synthetic peptides representing potential PKC phosphorylation sites, suggests that Ser(568) and Ser(674) of the C1 domain and Thr(931) of the C2 domain might act as substrates for PKC. We next created several full-length ACVI mutants in which one or more of the four likely PKC phosphorylation sites (Ser(10), Ser(568), Ser(674), and Thr(931)) were mutated to alanine. Simultaneous mutation of at least two of the three likely residues located in the N and C1 domains (Ser(10), Ser(568), and Ser(674)) was required to render ACVI variants completely insensitive to PKC treatment. In contrast, a single mutation of Thr(931) was sufficient to create a functional ACVI mutant that exhibited no detectable PKC-mediated inhibition, demonstrating the essentiality of Thr(931) to PKC-mediated regulation. Based on these results, we propose that the three cytosolic domains of ACVI might form a regulatory complex. Phosphorylation of this regulatory complex at different sites might induce a fine-tuning of the catalytic core complex and subsequently lead to alternation in the catalytic activity of ACVI. PMID- 11877399 TI - Crystal structure of binary and ternary complexes of serine hydroxymethyltransferase from Bacillus stearothermophilus: insights into the catalytic mechanism. AB - Serine hydroxymethyltransferase (SHMT), a member of the alpha-class of pyridoxal phosphate-dependent enzymes, catalyzes the reversible conversion of serine to glycine and tetrahydrofolate to 5,10-methylene tetrahydrofolate. We present here the crystal structures of the native enzyme and its complexes with serine, glycine, glycine, and 5-formyl tetrahydrofolate (FTHF) from Bacillus stearothermophilus. The first structure of the serine-bound form of SHMT allows identification of residues involved in serine binding and catalysis. The SHMT serine complex does not show any significant conformational change compared with the native enzyme, contrary to that expected for a conversion from an "open" to "closed" form of the enzyme. However, the ternary complex with FTHF and glycine shows the reported conformational changes. In contrast to the Escherichia coli enzyme, this complex shows asymmetric binding of the FTHF to the two monomers within the dimer in a way similar to the murine SHMT. Comparison of the ternary complex with the native enzyme reveals the structural basis for the conformational change and asymmetric binding of FTHF. The four structures presented here correspond to the various reaction intermediates of the catalytic pathway and provide evidence for a direct displacement mechanism for the hydroxymethyl transfer rather than a retroaldol cleavage. PMID- 11877400 TI - Cleavage of the xylosyl serine linkage between a core peptide and a glycosaminoglycan chain by cellulases. AB - We previously found that endo-beta-xylosidase from Patinopecten is an endo-type glycosidase that cleaves the xylosyl serine linkage between a glycosaminoglycan chain and its core protein (Takagaki, K., Kon, A., Kawasaki, H., Nakamura, T., Tamura, S., and Endo, M. (1990) J. Biol. Chem. 265, 854-860). Screening for endo beta-xylosidase activity in several cellulases detected this activity in the enzymes from Aspergillus niger, Penicillium funiculosum, Trichoderma reesei, Trichoderma viride, and Irpex lacteus. The cellulase derived from A. niger was purified, and its molecular weight was determined to be 26,000 by SDS-PAGE. Examination of the specificity of the cellulase revealed that 1) the enzyme acts on the linkage region (xylosyl serine) between a core peptide and a glycosaminoglycan chain; 2) enzymatic activity is greater with shorter glycosaminoglycan chains; 3) the enzyme readily hydrolyzes the linkage in glycosaminoglycan peptides, but intact proteoglycan is cleaved only slowly; and 4) the activity is unaffected by the glycosaminoglycan component (chondroitin sulfate, dermatan sulfate, and heparan sulfate). Judging from these enzymatic characteristics, this cellulase is different from the endo-beta-xylosidase of Patinopecten. We believe that this cellulase will become a useful tool in the further development of glycotechnology, because, like the endo-beta-xylosidase of Patinopecten, it enables the release of intact glycosaminoglycans from glycosaminoglycan peptides. PMID- 11877401 TI - Rapid phosphorylation of heterogeneous nuclear ribonucleoprotein C1/C2 in response to physiologic levels of hydrogen peroxide in human endothelial cells. AB - Hydrogen peroxide (H(2)O(2)) has been implicated as a signaling agent in numerous signal transduction pathways in mammalian cells. However, to date, no sensor for low concentrations (<10 microm) of H(2)O(2) has been identified. Using a functional proteomic approach, nuclear extracts from human umbilical vein endothelial cells were analyzed by two-dimensional PAGE with or without prior treatment with a low concentration of H(2)O(2). A protein doublet with a molecular mass of 39-41 kDa and a pI of approximately 5.0 was observed to be consistently altered by the treatment. Using proteolytic peptide mass fingerprinting, the protein was identified as heterogeneous nuclear ribonucleoprotein C1/C2, a nuclear restricted, pre-mRNA-binding protein. Upon two dimensional PAGE, each heterogeneous nuclear ribonucleoprotein-C splice form was present as multiple spots because of differing levels of phosphorylation. Upon treatment with H(2)O(2), there was an increase in phosphorylation at 10-20 min, which partially reversed by 30 min. Subsequently, at 60 min after treatment, a population of unphosphorylated protein was transiently present. The effects were observed with as little as 1 microm H(2)O(2) and were maximal with 5-8 microm H(2)O(2). The H(2)O(2)-stimulated phosphorylation was inhibited by catalase, but not by the transcriptional inhibitor actinomycin D. PMID- 11877402 TI - Cytotoxic necrotizing factor-1 contributes to Escherichia coli K1 invasion of the central nervous system. AB - Escherichia coli K1 invasion of brain microvascular endothelial cells (BMECs) is a prerequisite for penetration into the central nervous system and requires actin cytoskeletal rearrangements. Here, we demonstrate that E. coli K1 invasion of BMECs requires RhoA activation. In addition, we show that cytotoxic necrotizing factor-1 (CNF1) contributes to E. coli K1 invasion of brain endothelial cells in vitro and traversal of the blood-brain barrier in the experimental hematogenous meningitis animal model. These in vitro and in vivo effects of CNF1 were dependent upon RhoA activation as shown by (a) decreased invasion and RhoA activation with the Delta cnf1 mutant of E. coli K1 and (b) restoration of invasion frequency of the Delta cnf1 mutant to the level of the parent E. coli K1 strain in BMECs with constitutively active RhoA. In addition, CNF1-enhanced E. coli invasion of brain endothelial cells and stress fiber formation were independent of focal adhesion kinase and phosphatidylinositol 3-kinase activation. This is the first demonstration that CNF1 contributes to E. coli K1 invasion of BMECs. PMID- 11877403 TI - Quality control in the yeast secretory pathway: a misfolded PMA1 H+-ATPase reveals two checkpoints. AB - The yeast plasma-membrane H(+)-ATPase, encoded by PMA1, is delivered to the cell surface via the secretory pathway and has recently emerged as an excellent system for identifying quality control mechanisms along the pathway. In the present study, we have tracked the biogenesis of Pma1-G381A, a misfolded mutant form of the H(+)-ATPase. Although this mutant ATPase is arrested transiently in the peripheral endoplasmic reticulum, it does not become a substrate for endoplasmic reticulum-associated degradation nor does it appear to stimulate an unfolded protein response. Instead, Pma1-G381A accumulates in Kar2p-containing vesicular tubular clusters that resemble those previously described in mammalian cells. Like their mammalian counterparts, the yeast vesicular-tubular clusters may correspond to specific exit ports from the endoplasmic reticulum, since Pma1 G381A eventually escapes from them (still in a misfolded, trypsin-sensitive form) to reach the plasma membrane. By comparison with wild-type ATPase, Pma1-G381A spends a short half-life at the plasma membrane before being removed and sent to the vacuole for degradation in a process that requires both End4p and Pep4p. Finally, in a separate set of experiments, Pma1-G381A was found to impose its phenotype on co-expressed wild-type ATPase, transiently retarding the wild-type protein in the ER and later stimulating its degradation in the vacuole. Both effects serve to lower the steady-state amount of wild-type ATPase in the plasma membrane and, thus, can explain the co-dominant genetic behavior of the G381A mutation. Taken together, the results of this study establish Pma1-G381A as a useful new probe for the yeast secretory system. PMID- 11877404 TI - Diffusional mobility of the cystic fibrosis transmembrane conductance regulator mutant, delta F508-CFTR, in the endoplasmic reticulum measured by photobleaching of GFP-CFTR chimeras. AB - Mutations in the cystic fibrosis transmembrane conductance regulator protein (CFTR) cause cystic fibrosis. The most common disease-causing mutation, DeltaF508, is retained in the endoplasmic reticulum (ER) and is unable to function as a plasma membrane chloride channel. To investigate whether the ER retention of DeltaF508-CFTR is caused by immobilization and/or aggregation, we have measured the diffusional mobility of green fluorescent protein (GFP) chimeras of wild type (wt)-CFTR and DeltaF508-CFTR by fluorescence recovery after photobleaching. GFP-labeled DeltaF508-CFTR was localized in the ER and wt-CFTR in the plasma membrane and intracellular membranes in transfected COS7 and Chinese hamster ovary K1 cells. Both chimeras localized to the ER after brefeldin A treatment. Spot photobleaching showed that CFTR diffusion (diffusion coefficient approximately 10(-9) cm(2)/s) was not significantly slowed by the DeltaF508 mutation and that nearly all wt-CFTR and DeltaF508-CFTR diffused throughout the ER without restriction. Stabilization of molecular chaperone interactions by ATP depletion produced remarkable DeltaF508-CFTR immobilization ( approximately 50%) and slowed diffusion (6.5 x 10(-10) cm(2)/s) but had little effect on wt-CFTR. Fluorescence depletion experiments revealed that the immobilized DeltaF508-CFTR in ATP-depleted cells remained in an ER pattern. The mobility of wt-CFTR and DeltaF508-CFTR was reduced by maneuvers that alter CFTR processing or interactions with molecular chaperones, including tunicamycin, geldanamycin, and lactacystin. Photobleaching of the fluorescent ER lipid diOC(4)(3) showed that neither ER restructuring nor fragmentation during these maneuvers was responsible for the slowing and immobilization of CFTR. These results suggest that (a) the ER retention of DeltaF508-CFTR is not due to restricted ER mobility, (b) the majority of DeltaF508-CFTR is not aggregated or bound to slowly moving membrane proteins, and (c) DeltaF508-CFTR may interact to a greater extent with molecular chaperones than does wt-CFTR. PMID- 11877406 TI - Substitution of the autophosphorylation site Thr516 with a negatively charged residue confers constitutive activity to mouse 3-phosphoinositide-dependent protein kinase-1 in cells. AB - 3-Phosphoinositide-dependent protein kinase-1 (PDK-1)is a serine/threonine kinase that has been found to phosphorylate and activate several members of the AGC protein kinase family including protein kinase B (Akt), p70 S6 kinase, and protein kinase Czeta. However, the mechanism(s) by which PDK-1 is regulated remains unclear. Here we show that mouse PDK-1 (mPDK-1) undergoes autophosphorylation in vitro on both serine and threonine residues. In addition, we have identified Ser(399) and Thr(516) as the major mPDK-1 autophosphorylation sites in vitro. Furthermore, we have found that these two residues, as well as Ser(244) in the activation loop, are phosphorylated in cells and demonstrated that Ser(244) is a major in vivo phosphorylation site. Abolishment of phosphorylation at Ser(244), but not at Ser(399) or Thr(516), led to a significant decrease of mPDK-1 autophosphorylation and kinase activity in vitro, indicating that autophosphorylation at Ser(399) or Thr(516) is not essential for mPDK-1 autokinase activity. However, overexpression of mPDK-1(T516E), but not of mPDK-1(S244E) or mPDK-1(S399D), in Chinese hamster ovary and HEK293 cells was sufficient to induce Akt phosphorylation at Thr(308) to a level similar to that of insulin stimulation. Furthermore, this increase in phosphorylation was independent of the Pleckstrin homology domain of Akt. Taken together, our results suggest that mPDK-1 undergoes autophosphorylation at multiple sites and that this phosphorylation may be essential for PDK-1 to interact with and phosphorylate its downstream substrates in vivo. PMID- 11877405 TI - A tale of two controversies: defining both the role of peroxidases in nitrotyrosine formation in vivo using eosinophil peroxidase and myeloperoxidase deficient mice, and the nature of peroxidase-generated reactive nitrogen species. AB - Nitrotyrosine is widely used as a marker of post-translational modification by the nitric oxide ((.)NO, nitrogen monoxide)-derived oxidant peroxynitrite (ONOO( )). However, since the discovery that myeloperoxidase (MPO) and eosinophil peroxidase (EPO) can generate nitrotyrosine via oxidation of nitrite (NO(2)(-)), several questions have arisen. First, the relative contribution of peroxidases to nitrotyrosine formation in vivo is unknown. Further, although evidence suggests that the one-electron oxidation product, nitrogen dioxide ((*)NO(2)), is the primary species formed, neither a direct demonstration that peroxidases form this gas nor studies designed to test for the possible concomitant formation of the two-electron oxidation product, ONOO(-), have been reported. Using multiple distinct models of acute inflammation with EPO- and MPO-knockout mice, we now demonstrate that leukocyte peroxidases participate in nitrotyrosine formation in vivo. In some models, MPO and EPO played a dominant role, accounting for the majority of nitrotyrosine formed. However, in other leukocyte-rich acute inflammatory models, no contribution for either MPO or EPO to nitrotyrosine formation could be demonstrated. Head-space gas analysis of helium-swept reaction mixtures provides direct evidence that leukocyte peroxidases catalytically generate (*)NO(2) formation using H(2)O(2) and NO(2)(-) as substrates. However, formation of an additional oxidant was suggested since both enzymes promote NO(2)(-)-dependent hydroxylation of targets under acidic conditions, a chemical reactivity shared with ONOO(-) but not (*)NO(2). Collectively, our results demonstrate that: 1) MPO and EPO contribute to tyrosine nitration in vivo; 2) the major reactive nitrogen species formed by leukocyte peroxidase-catalyzed oxidation of NO(2)(-) is the one-electron oxidation product, (*)NO(2); 3) as a minor reaction, peroxidases may also catalyze the two-electron oxidation of NO(2)(-), producing a ONOO(-)-like product. We speculate that the latter reaction generates a labile Fe-ONOO complex, which may be released following protonation under acidic conditions such as might exist at sites of inflammation. PMID- 11877407 TI - Similar structural basis for membrane localization and protein priming by an RNA dependent RNA polymerase. AB - Protein primers are used to initiate genomic synthesis of several RNA and DNA viruses, although the structural details of the primer-polymerase interactions are not yet known. Poliovirus polymerase binds with high affinity to the membrane bound viral protein 3AB but uridylylates only the smaller peptide 3B in vitro. Mutational analysis of the polymerase identified four surface residues on the three-dimensional structure of poliovirus polymerase whose wild-type identity is required for 3AB binding. These mutants also decreased 3B uridylylation, arguing that the binding sites for the membrane tether and the protein primer overlap. Mutation of flanking residues between the 3AB binding site and the polymerase active site specifically decreased 3B uridylylation, likely affecting steps subsequent to binding. The physical overlap of sites for protein priming and membrane association should facilitate replication initiation in the membrane associated complex. PMID- 11877408 TI - The promoter of the human proliferating cell nuclear antigen gene is not sufficient for cell cycle-dependent regulation in organotypic cultures of keratinocytes. AB - The proliferating cell nuclear antigen (PCNA) is essential for DNA replication of mammalian cells and their small DNA tumor viruses. The mechanism of the cell cycle-dependent regulation of the human PCNA promoter is not clear despite extensive investigations. In this report, we employed organotypic cultures of primary human keratinocytes, which closely resemble native skin comprising both proliferating and postmitotic, differentiated cells, to examine the cell cycle dependent regulation of the human PCNA gene (hPCNA) in the absence or presence of the human papillomavirus type 18 (HPV-18) E7 protein. HPV-18 E7 promotes S phase re-entry in post-mitotic differentiated keratinocytes by abrogating the transcription repression of E2F transcription factors by the retinoblastoma susceptibility protein, pRb. We demonstrated that E7 reactivated the transcription of the endogenous hPCNA in differentiated keratinocytes. In contrast, with or without E7, the expression of a transduced hPCNA promoter driven reporter did not correlate with that of the endogenous hPCNA gene in either proliferating or differentiated cells. Moreover, in Chinese hamster ovary and L-cells, HPV E7 and the adenovirus E1A protein repressed the transduced hPCNA promoter, but both activated an extended promoter construct spanning the first intron. Mutations of two E2F sites in the intron reduced the basal activity and abolished the response to E7 or E1A. Promoter repression or activation required the CR2 domain of E7 and, to a lesser extent, CR1 as well. However, in organotypic cultures, this extended promoter construct failed to recapitulate the cell cycle-dependent regulation of the endogenous hPCNA gene. Only when a full length Myc-tagged hPCNA spanning the 5' promoter and all exons and introns was used was the native pattern of expression largely restored, indicative of the complexity of its regulation. PMID- 11877410 TI - An iron-sulfur cluster in the family 4 uracil-DNA glycosylases. AB - The 25-kDa Family 4 uracil-DNA glycosylase (UDG) from Pyrobaculum aerophilum has been expressed and purified in large quantities for structural analysis. In the process we observed it to be colored and subsequently found that it contained iron. Here we demonstrate that P. aerophilum UDG has an iron-sulfur center with the EPR characteristics typical of a 4Fe4S high potential iron protein. Interestingly, it does not share any sequence similarity with the classic iron sulfur proteins, although four cysteines (which are strongly conserved in the thermophilic members of Family 4 UDGs) may represent the metal coordinating residues. The conservation of these residues in other members of the family suggest that 4Fe4S clusters are a common feature. Although 4Fe4S clusters have been observed previously in Nth/MutY DNA repair enzymes, this is the first observation of such a feature in the UDG structural superfamily. Similar to the Nth/MutY enzymes, the Family 4 UDG centers probably play a structural rather than a catalytic role. PMID- 11877409 TI - Mutations in the occQ operator that decrease OccR-induced DNA bending do not cause constitutive promoter activity. AB - OccR is a LysR-type transcriptional regulator of Agrobacterium tumefaciens that positively regulates the octopine catabolism operon of the Ti plasmid. Positive control of the occ genes occurs in response to octopine, a metabolite released from plant tumors. Octopine causes DNA-bound OccR to undergo a conformational change from an inactive to an active state; this change is marked by a decrease in footprint length from 55 to 45 nucleotides as well as a relaxation of a high angle DNA bend. In this study, we first used gel filtration chromatography to show that OccR is dimeric in solution, and we used gel shift assays to show that OccR is tetrameric when bound to DNA. We then created a series of site-directed mutations in the OccR-binding site. Some mutations were designed to lock OccR-DNA complexes into a conformation resembling the inactive conformation, whereas other mutations were designed to lock complexes into the active conformation. These mutations altered the conformation of OccR-DNA complexes and their responses to octopine in ways that we had predicted. As expected, operator mutations that locked complexes into a conformation having a long footprint and a high angle DNA bend blocked activation by octopine in vivo. Surprisingly, however, mutations that lock OccR into a short footprint and low angle DNA bend failed to cause the protein to function constitutively. Furthermore, some of the latter mutations interfered with activation by octopine. We conclude that locking OccR into a conformation having a short footprint is not sufficient to cause constitutive activation, and octopine must cause at least one additional conformational change in the protein. PMID- 11877411 TI - The structure of a binary complex between a mammalian mevalonate kinase and ATP: insights into the reaction mechanism and human inherited disease. AB - Mevalonate kinase catalyzes the ATP-dependent phosphorylation of mevalonic acid to form mevalonate 5-phosphate, a key intermediate in the pathways of isoprenoids and sterols. Deficiency in mevalonate kinase activity has been linked to mevalonic aciduria and hyperimmunoglobulinemia D/periodic fever syndrome (HIDS). The crystal structure of rat mevalonate kinase in complex with MgATP has been determined at 2.4-A resolution. Each monomer of this dimeric protein is composed of two domains with its active site located at the domain interface. The enzyme bound ATP adopts an anti conformation, in contrast to the syn conformation reported for Methanococcus jannaschii homoserine kinase. The Mg(2+) ion is coordinated to both beta- and gamma-phosphates of ATP and side chains of Glu(193) and Ser(146). Asp(204) is making a salt bridge with Lys(13), which in turn interacts with the gamma-phosphate. A model of mevalonic acid can be placed near the gamma-phosphoryl group of ATP; thus, the C5 hydroxyl is located within 4 A from Asp(204), Lys(13), and the gamma-phosphoryl of ATP. This arrangement of residues strongly suggests: 1) Asp(204) abstracts the proton from C5 hydroxyl of mevalonate; 2) the penta-coordinated gamma-phosphoryl group may be stabilized by Mg(2+), Lys(13), and Glu(193); and 3) Lys(13) is likely to influence the pK(a) of the C5 hydroxyl of the substrate. V377I and I268T are the most common mutations found in patients with HIDS. Val(377) is located over 18 A away from the active site and a conservative replacement with Ile is unlikely to yield an inactive or unstable protein. Ile-268 is located at the dimer interface, and its Thr substitution may disrupt dimer formation. PMID- 11877412 TI - Mapping of a ligand-binding site for the human thromboxane A2 receptor protein. AB - The human thromboxane A(2) (TP) receptor, a member of the G protein-coupled receptor superfamily, consists of seven transmembrane segments. Attempts to elucidate the specific segment(s) that define the receptor ligand-binding pocket have produced less than definitive and sometimes conflicting results. On this basis, the present work identified an amino acid sequence of the TP receptor that is directly involved in ligand binding. Mapping of this domain was confirmed by two separate approaches: photoaffinity labeling and site-specific antibodies. The newly synthesized, biotinylated photoaffinity probe, SQBAzide, was first shown to specifically label TP receptor protein. Sequential digestion of this protein with CNBr/trypsin revealed photolabeling of a 2.9-kDa peptide. Using anti-peptide antibodies directed against different regions of the receptor protein, it was established that this peptide represents the predicted cleavage product for CNBr/trypsin and corresponds to amino acids Arg(174)-Met(202) of the receptor protein. Furthermore, antibody screening revealed that inhibition of the amino acid region Cys(183)-Asp(193) was critical for radioligand binding and platelet aggregation, whereas inhibition of Gly(172)-Cys(183) was not. Collectively these findings provide evidence that ligands interact with amino acids contained within the C-terminal portion of the third extracellular domain (ED3) of the receptor protein. This information should be of significant value in the study of TP receptor structure and signaling. PMID- 11877413 TI - Nuclear factor 1-C2 contributes to the tissue-specific activation of a milk protein gene in the differentiating mammary gland. AB - Members of the nuclear factor 1 (NF1) transcription factor family have been postulated to be involved in the regulation of milk genes. In this work we have been able to identify the splice variant NF1-C2 as an important member of a tissue-specific activating complex that regulates the milk gene encoding carboxyl ester lipase (CEL). Mutation of the NF1-binding site in the CEL gene promoter results in a drastic reduction of the gene expression to about 15% in mammary epithelial cells. Furthermore, we demonstrate that the NF1-C2 protein interacts with a higher affinity to the NF1-binding site in the CEL gene promoter than other NF1 family members do and that NF1-C2 in the mouse mammary gland is a phosphorylated protein. During development of the mouse mammary gland, binding of NF1-C2 to the CEL gene promoter is induced at midpregnancy, in correlation with the induction of CEL gene expression. The fact that the NF1-C2 involving complex remains throughout the lactation period and decreases during the weaning period, when the CEL gene is down-regulated, supports its importance in the regulation of CEL gene expression. To our knowledge, this is the first report identifying a specific, endogenously expressed NF1 isoform to be involved in the tissue specific activation of a gene. PMID- 11877414 TI - Amyloid precursor protein family-induced neuronal death is mediated by impairment of the neuroprotective calcium/calmodulin protein kinase IV-dependent signaling pathway. AB - The aberrant metabolism of beta-amyloid precursor protein (APP) and the progressive deposition of its derived fragment beta-amyloid peptide are early and constant pathological hallmarks of Alzheimer's disease. Because APP is able to function as a cell surface receptor, we investigated here whether a disruption of the normal function of APP may contribute to the pathogenic mechanisms in Alzheimer's disease. To this aim, we generated a specific chicken polyclonal antibody directed against the extracellular domain of APP, which is common with the beta-amyloid precursor-like protein type 2. Exposure of cultured cortical neurons to this antibody (APP-Ab) induced cell death preceded by neurite degeneration, oxidative stress, and nuclear condensation. Interestingly, caspase 3-like protease was not activated in this neurotoxic action suggesting a different mode of cell death than classical apoptosis. Further analysis of the molecular mechanisms revealed a calpain- and calcineurin-dependent proteolysis of the neuroprotective calcium/calmodulin-dependent protein kinase IV and its nuclear target protein cAMP responsive element binding protein. These effects were abolished by the G protein inhibitor pertussis toxin, strongly suggesting that APP binding operates via a GTPase-dependent pathway to cause neuronal death. PMID- 11877415 TI - Early endosomal regulation of Smad-dependent signaling in endothelial cells. AB - Transforming growth factor beta (TGFbeta) receptors require SARA for phosphorylation of the downstream transducing Smad proteins. SARA, a FYVE finger protein, binds to membrane lipids suggesting that activated receptors may interact with downstream signaling molecules at discrete endocytic locations. In the present study, we reveal a critical role for the early endocytic compartment in regulating Smad-dependent signaling. Not only is SARA localized on early endosomes, but also its minimal FYVE finger sequence is sufficient for early endosomal targeting. Expression of a SARA mutant protein lacking the FYVE finger inhibits downstream activin A signaling in endothelial cells. Moreover, a dominant-negative mutant of Rab5, a crucial protein for early endosome dynamics, causes phosphorylation and nuclear translocation of Smads leading to constitutive (i.e. ligand independent) transcriptional activation of a Smad-dependent promoter in endothelial cells. As inhibition of endocytosis using the K44A negative mutant of dynamin and RN-tre did not lead to activation of Smad-dependent transcription, the effects of the dominant-negative Rab5 are likely to be a consequence of altered membrane trafficking of constitutively formed TGFbeta/activin type I/II receptor complexes at the level of early endosomes. The results suggest an important interconnection between early endosomal dynamics and TGFbeta/activin signal transduction pathways. PMID- 11877416 TI - Identification of a substrate recognition site on Ubc9. AB - Human Ubc9 is homologous to ubiquitin-conjugating enzymes. However, instead of conjugating ubiquitin, it conjugates a ubiquitin homologue, small ubiquitin-like modifier 1 (SUMO-1), also known as UBL1, GMP1, SMTP3, PIC1, and sentrin. The SUMO 1 conjugation pathway is very similar to that of ubiquitin with regard to the primary sequences of the ubiquitin-activating enzymes (E1), the three-dimensional structures of the ubiquitin-conjugating enzymes (E2), and the chemistry of the overall conjugation pathway. The interaction of substrates with Ubc9 has been studied using NMR spectroscopy. Peptides with sequences that correspond to those of the SUMO-1 conjugation sites from p53 and c-Jun both bind to a surface adjacent to the active site Cys93 of human Ubc9, which has been previously shown to include residues that demonstrate the most significant dynamics on the microsecond to millisecond time scale. Mutations in this region, Q126A, Q130A, A131D, E132A, Y134A, and T135A, were constructed to evaluate the role of these residues in SUMO-1 conjugation. These alterations have significant effects on the conjugation of SUMO-1 with the target proteins p53, E1B, and promyelocytic leukemia protein and define a substrate binding site on Ubc9. Furthermore, the SUMO-1 conjugation site of p53 does not form any defined secondary structure when either free or bound to Ubc9. This suggests that a defined secondary structure at SUMO-1 conjugation sites in target proteins is not necessary for recognition and conjugation by the SUMO-1 pathway. PMID- 11877417 TI - Ligand discrimination by TPR domains. Relevance and selectivity of EEVD recognition in Hsp70 x Hop x Hsp90 complexes. AB - Protein-protein interaction modules containing so-called tetratricopeptide repeats (TPRs) mediate the assembly of Hsp70/Hsp90 multi-chaperone complexes. The TPR1 and TPR2A domains of the Hsp70/Hsp90 adapter protein p60/Hop specifically bind to short peptides corresponding to the C-terminal tails of Hsp70 and Hsp90, respectively, both of which contain the highly conserved sequence motif EEVD COOH. Here, we quantitatively assessed the contribution of TPR-mediated peptide recognition to Hsp70.Hop.Hsp90 complex formation. The interaction of TPR2A with the C-terminal pentapeptide of Hsp90 (MEEVD) is identified as the core contact for Hop binding to Hsp90. (In peptide sequences, italics are used to highlight residues specific for Hsp70 or Hsp90.) In contrast, formation of the Hsp70.Hop complex depends not only on recognition of the C-terminal Hsp70 heptapeptide (PTIEEVD) by TPR1 but also on additional contacts between Hsp70 and Hop. The sequence motifs for TPR1 and TPR2A binding were defined by alanine scanning of the C-terminal octapeptides of Hsp70 and Hsp90 and by screening of combinatorial peptide libraries. Asp0 and Val-1 of the EEVD motif are identified as general anchor residues, but the highly conserved glutamates of the EEVD sequence, which are critical in Hsp90 binding by TPR2A, do not contribute appreciably to the interaction of Hsp70 with TPR1. Rather, TPR1 prefers hydrophobic amino acids in these positions. Moreover, the TPR domains display a pronounced tendency to interact preferentially with hydrophobic aliphatic and aromatic side chains in positions -4 and -6 of their respective peptide ligands. Ile-4 in Hsp70 and Met-4 in Hsp90 are most important in determining the specific binding of TPR1 and TPR2A, respectively. PMID- 11877418 TI - Protein inhibitors of activated STAT resemble scaffold attachment factors and function as interacting nuclear receptor coregulators. AB - Protein inhibitor of activated STAT1 (PIAS1) functions as a nuclear receptor coregulator and is expressed in several cell types of human testis. However, the mechanism of PIAS1 coregulation is unknown. We report here that PIAS1 has characteristics of a scaffold attachment protein. PIAS1 localized in nuclei in a speckled pattern and bound A-T-rich double-stranded DNA, a function of scaffold attachment proteins in chromatin regions of active transcription. DNA binding was dependent on a 35-amino acid sequence conserved among members of the PIAS family and in scaffold attachment proteins. The PIAS family also bound the androgen receptor DNA binding domain, and binding required the second zinc finger of this domain. PIAS1 contained an intrinsic activation domain but had bi-directional effects on androgen receptor transactivation; lower expression levels inhibited and higher levels increased transactivation in CV1 cells. Other PIAS family members also had dose-dependent effects on transactivation, but they were in a direction opposite to those of PIAS1. When coexpressed with PIAS1, other PIAS family members counteracted PIAS1 coregulation of androgen receptor transactivation. The interaction of PIAS1 with other members of the PIAS family suggests a transcription coregulatory mechanism involving a multicomponent PIAS nuclear scaffold. PMID- 11877419 TI - Ultraviolet light inhibits translation through activation of the unfolded protein response kinase PERK in the lumen of the endoplasmic reticulum. AB - Exposure to ultraviolet light can cause inflammation, premature skin aging, and cancer. UV irradiation alters the expression of multiple genes that encode functions to repair DNA damage, arrest cell growth, and induce apoptosis. In addition, UV irradiation inhibits protein synthesis, although the mechanism is not known. In this report, we show that UV irradiation induces phosphorylation of eukaryotic translation initiation factor 2 on the alpha-subunit (eIF2alpha) and inhibits protein synthesis in a dosage- and time-dependent manner. The UV-induced phosphorylation of eIF2alpha was prevented by the overexpression of a non phosphorylatable mutant of eIF2alpha (S51A). PERK is an eIF2alpha protein kinase localized to the endoplasmic reticulum that is activated by the accumulation of unfolded proteins in the endoplasmic reticulum. Expression of trans-dominant negative mutants of PERK also prevented eIF2alpha phosphorylation upon UV treatment and protected from the associated translation attenuation. The luminal domain of dominant-negative mutant PERK formed heterodimers with endogenous PERK to inhibit the PERK signaling pathway. In contrast, eIF2alpha phosphorylation was not inhibited by overexpression of a trans-dominant-negative mutant kinase, PKR, supporting the theory that UV-induced eIF2alpha phosphorylation is specifically mediated by PERK. These results support a novel mechanism by which UV irradiation regulates translation via an endoplasmic reticulum-stress signaling pathway. PMID- 11877420 TI - Tyrosine phosphorylation of the beta-amyloid precursor protein cytoplasmic tail promotes interaction with Shc. AB - beta-Amyloid precursor protein (APP) is a widely expressed transmembrane protein of unknown function that is involved in the pathogenesis of Alzheimer's disease. The cytoplasmic tail of APP interacts with phosphotyrosine binding (PTB) domain containing proteins (Fe65, X11, mDab-1, and JIP-1) and may modulate gene expression and apoptosis. We now identify Shc A and Shc C, PTB-containing adapter proteins that signal to cellular differentiation and survival pathways, as novel APP-interacting proteins. The APP cytoplasmic tail contains a PTB-binding motif (Y(682)ENPTY(687)) that, when phosphorylated on Tyr(682), precipitated the PTB domain of Shc A and Shc C, as well as endogenous full-length Shc A. APP and Shc C were physically associated in adult mouse brain homogenates. Increase in phosphorylation of APP by overexpression of the nerve growth factor receptor Trk A in 293T cells promoted the interaction of transfected APP and endogenous Shc A. Pervanadate treatment of N2a neuroblastoma cells resulted in tyrosine phosphorylation and association of endogenous APP and Shc A. Thus, APP and Shc proteins interact in vitro, in cells, and in the mouse brain. Tyrosine phosphorylation of APP may promote the interaction with Shc proteins. PMID- 11877421 TI - A kinetic model of intermediate formation during assembly of cholera toxin B subunit pentamers. AB - Cholera toxin is the most important virulence factor produced by Vibrio cholerae. The pentameric B-subunit of the toxin can bind to GM1-ganglioside receptors, leading to toxin entry into mammalian cells. Here, the in vitro disassembly and reassembly of CtxB(5) (the B subunit pentamer of cholera toxin) is investigated. When CtxB(5) was acidified at pH 1.0 and then neutralized, the B-subunits disassembled and could no longer migrate as SDS-stable pentamers on polyacrylamide gels or be captured by GM1. However, continued incubation at neutral pH resulted in the B-subunits regaining the capacity to be detected by GM1 enzyme-linked immunosorbent assay (t(12) approximately 8 min) and to migrate as SDS-stable pentamers (t(12) approximately 15 min). Time-dependent changes in Trp fluorescence intensity during B-subunit reassembly occurred with a half-time of approximately 8 min, similar to that detected by GM1 enzyme-linked immunosorbent assay, suggesting that both methods monitor earlier events than B pentamer formation alone. Based on the Trp fluorescence intensity measurements, a kinetic model of the pathway of CtxB(5) reassembly was generated that depended on trans to cis isomerization of Pro-93 to give an interface capable of subunit subunit interaction. The model suggests formation of intermediates in the reaction, and these were successfully detected by glutaraldehyde cross-linking. PMID- 11877422 TI - Amyloid-like fibril formation in an all beta-barrel protein involves the formation of partially structured intermediate(s). AB - In the present study, we demonstrate the thermal induced amyloid formation in a beta-barrel protein, such as the acidic fibroblast growth factor from Notopthalmus viridescens (nFGF-1). Fibril formation in nFGF-1 is observed to occur maximally at 65 degrees C. Electron microscope analysis of the thermal induced fibrils of nFGF-1 shows that they are filamentous with an average diameter of about 20 nm. X-ray diffraction analysis reveals that the thermal induced fibrils of nFGF-1 have a typical "cross-beta" structure with the beta strands perpendicular to the fibril axis. By using a variety of biophysical techniques including multidimensional NMR, we demonstrate that fibril formation involves the formation of a partially structured intermediate(s) in the thermal unfolding pathway of the protein (nFGF-1). Results of the anilino-8-napthalene sulfonate binding experiments indicate that fibril formation occurs due to the coalescence of the protein (in the intermediate state(s)) through the solvent exposed non-polar surface(s). In this study, we also demonstrate that organic osmolytes, such as proline, can efficiently prevent the thermal induced amyloid formation in nFGF-1. Proline is found to stabilize the native conformation of the protein. The addition, proline is observed to increase the cooperativity of the unfolding (native <--> denatured) reaction and consequently decrease the population of the "sticky" thermal equilibrium intermediate(s) responsible for the fibril formation. PMID- 11877423 TI - Composition and function of AP-1 transcription complexes during muscle cell differentiation. AB - The role of activating protein-1 (AP-1) in muscle cells is currently equivocal. While some studies propose that AP-1 is inhibitory for myogenesis, others implicate a positive role in this process. We tested whether this variation may be due to different properties of the AP-1 subunit composition in differentiating cells. Using Western analysis we show that c-Jun, Fra-2, and JunD are expressed throughout the time course of differentiation. Phosphatase assays indicate that JunD and Fra-2 are phosphorylated in muscle cells and that at least two isoforms of each are expressed in muscle cells. Electrophoretic mobility shift assays combined with antibody supershifts indicate the appearance of Fra-2 as a major component of the AP-1 DNA binding complex in differentiating cells. In this context it appears that Fra-2 heterodimerizes with c-Jun and JunD. Studying the c jun enhancer in reporter gene assays we observed that the muscle transcription factors MEF2A and MyoD can contribute to robust transcriptional activation of the c-jun enhancer. In differentiating muscle cells mutation of the MEF2 site reduces transactivation of the c-jun enhancer and MEF2A is the predominant MEF2 isoform binding to this cis element. Transcriptional activation of an AP-1 site containing reporter gene (TRE-Luc) is enhanced under differentiation conditions compared with growth conditions in C2C12 muscle cells. Further studies indicate that Fra-2 containing AP-1 complexes can transactivate the MyoD enhancer/promoter. Thus, an AP-1 complex containing Fra-2 and c-Jun or JunD is consistent with muscle differentiation, indicating that AP-1 function during myogenesis is dependent on its subunit composition. PMID- 11877424 TI - Dynamin is a minibrain kinase/dual specificity Yak1-related kinase 1A substrate. AB - The minibrain kinase (Mnbk)/dual specificity Yak 1-related kinase 1A (Dyrk1A) gene is implicated in the mental retardation associated with Down's syndrome. It encodes a proline-directed serine/threonine kinase whose function has yet to be defined. We have used a solid-phase Mnbk/Dyrk1A kinase assay to aid in the search for the cellular Mnbk/Dyrk1A substrates. The assay revealed that rat brain contains two cytosolic proteins, one with a molecular mass of 100 kDa and one with a molecular mass of 140 kDa, that were prominently phosphorylated by Mnbk/Dyrk1A. The 100-kDa protein was purified and identified as dynamin 1. The conclusion was further supported by evidence that a recombinant glutathione S transferase fusion protein containing dynamin isoform 1aa was phosphorylated by Mnbk/Dyrk1A. In addition to isoform 1aa, Mnbk/Dyrk1A also phosphorylated isoforms 1ab and 2aa but not human MxA protein when analyzed by the solid-phase kinase assay. Upon Mnbk/Dyrk1A phosphorylation, the interaction of dynamin 1 with the Src homology 3 domain of amphiphysin 1 was reduced. However, when Mnbk/Dyrk1A phosphorylation was allowed to proceed more extensively, the phosphorylation enhanced rather than reduced the binding of dynamin 1 to amphiphysin 1. The result suggests that Mnbk/Dyrk1A can play a dual role in regulating the interaction of dynamin 1 with amphiphysin 1. Mnbk/Dyrk1A phosphorylation also reduced the interaction of dynamin with endophilin 1, whereas the same phosphorylation enhanced the binding of dynamin 1 to Grb2. Nevertheless, the dual function of Mnbk/Dyrk1A phosphorylation was not observed for the interaction of dynamin 1 with endophilin 1 or Grb2. The interactions of dynamin with amphiphysin and endophilin are essential for the formation of endocytic complexes; our results suggest that Mnbk/Dyrk1A may function as a regulator controlling the assembly of endocytic apparatus. PMID- 11877425 TI - Point mutation in the first transmembrane region of the beta 2 subunit of the gamma--aminobutyric acid type A receptor alters desensitization kinetics of gamma -aminobutyric acid- and anesthetic-induced channel gating. AB - A conserved glycine residue in the first transmembrane (TM1) domain of the beta2 subunit has been identified to be involved with desensitization induced by gamma aminobutyric acid (GABA) and anesthetics. Recombinant GABA(A) receptors expressed in Sf9 cells were recorded using semi-fast agonist application. Upon direct activation by GABA or anesthetics, the main effect of the TM1 point mutation on the beta2 subunit (G219F) was to slow the time constant (tau) of desensitization. At GABA concentrations eliciting maximum currents, the corresponding median tau values were 0.87 s (25-75% interval (0.76; 1.04 s)), 0.93 s (0.76; 1.23 s), and 1.36 s (1.17; 1.57 s) for alpha1beta2gamma2, alpha1(G223F)beta2gamma2, and alpha1beta2(G219F)gamma2, respectively. The tau value for the beta2-mutant receptor was significantly longer than alpha1beta2gamma2 (p < 0.01) and alpha1(G223F)beta2gamma2 (p < 0.05). For pentobarbital-induced currents (500 microm), the corresponding median tau values were 1.36 s (0.81; 1.41 s), 1.47 s (1.31; 2.38 s), and 2.82 s (2.21; 5.56 s) for alpha1beta2gamma2, alpha1(G223F)beta2gamma2, and alpha1beta2(G219F)gamma2, respectively. The tau value for the beta2-mutant receptor was significantly longer than that for alpha1beta2gamma2 (p < 0.01). The present findings suggest that this TM1 glycine residue is critical for the rate at which desensitization occurs and that both GABA and intravenous anesthetics implement an analogous pathway for generating desensitization. PMID- 11877426 TI - Nerve growth factor-dependent activation of the small GTPase Rin. AB - The Rit and Rin proteins comprise a distinct and evolutionarily conserved subfamily of Ras-related small GTPases. Although we have defined a role for Rit mediated signal transduction in the regulation of cell proliferation and transformation, the function of Rin remains largely unknown. Because we demonstrate that Rin is developmentally regulated and expressed in adult neurons, we examined its role in neuronal signaling. In this study, we show that stimulation of PC6 cells with either epidermal growth factor or nerve growth factor (NGF) results in rapid activation of Rin. This activation correlates with the onset of Ras activation, and dominant-negative Ras completely inhibits Rin activation induced by NGF. Further examination of Ras-mediated Rin activation suggests that this process is dependent upon neuronally expressed regulatory factors. Expression of mutationally activated H-Ras fails to activate Rin in non neuronal cells, but results in potent stimulation of Rin-GTP levels in a variety of neuronal cell lines. Furthermore, although constitutively activated Rin does not induce neurite outgrowth on its own, both NGF-induced and oncogenic Ras induced neurite outgrowth were inhibited by the expression of dominant-negative Rin. Together, these studies indicate that Rin activation is a direct downstream effect of growth factor-dependent signaling in neuronal cells and suggest that Rin may function to transduce signals within the mature nervous system. PMID- 11877427 TI - Identification of target tissue glycosphingolipid receptors for uropathogenic, F1C-fimbriated Escherichia coli and its role in mucosal inflammation. AB - Bacterial adherence to mucosal cells is a key virulence trait of pathogenic bacteria. The type 1 fimbriae and the P-fimbriae of Escherichia coli have both been described to be important for the establishment of urinary tract infections. While P-fimbriae recognize kidney glycosphingolipids carrying the Galalpha4Gal determinant, type 1 fimbriae bind to the urothelial mannosylated glycoproteins uroplakin Ia and Ib. The F1C fimbriae are one additional type of fimbria correlated with uropathogenicity. Although it was identified 20 years ago its receptor has remained unidentified. Here we report that F1C-fimbriated bacteria selectively interact with two minor glycosphingolipids isolated from rat, canine, and human urinary tract. Binding-active compounds were isolated and characterized as galactosylceramide, and globotriaosylceramide, both with phytosphingosine and hydroxy fatty acids. Comparison with reference glycosphingolipids revealed that the receptor specificity is dependent on the ceramide composition. Galactosylceramide was present in the bladder, urethers, and kidney while globotriaosylceramide was present only in the kidney. Using a functional assay, we demonstrate that binding of F1C-fimbriated Escherichia coli to renal cells induces interleukin-8 production, thus suggesting a role for F1C-mediated attachment in mucosal defense against bacterial infections. PMID- 11877428 TI - Importance of C1B domain for lipid messenger-induced targeting of protein kinase C. AB - The molecular mechanisms by which arachidonic acid (AA) and ceramide elicit translocation of protein kinase C (PKC) were investigated. Ceramide translocated epsilonPKC from the cytoplasm to the Golgi complex, but with a mechanism distinct from that utilized by AA. Using fluorescence recovery after photobleaching, we showed that, upon treatment with AA, epsilonPKC was tightly associated with the Golgi complex; ceramide elicited an accumulation of epsilonPKC which was exchangeable with the cytoplasm. Stimulation with ceramide after AA converted the AA-induced Golgi complex staining to one elicited by ceramide alone; AA had no effect on the ceramide-stimulated localization. Using point mutants and deletions of epsilonPKC, we determined that the epsilonC1B domain was responsible for the ceramide- and AA-induced translocation. Switch chimeras, containing the C1B from epsilonPKC in the context of deltaPKC (delta(epsilonC1B)) and vice versa (epsilon(deltaC1B)), were generated and tested for their translocation in response to ceramide and AA. delta(epsilonC1B) translocated upon treatment with both ceramide and AA; epsilon(deltaC1B) responded only to ceramide. Thus, through the C1B domain, AA and ceramide induce different patterns of epsilonPKC translocation and the C1B domain defines the subtype specific sensitivity of PKCs to lipid second messengers. PMID- 11877429 TI - Toll-like receptor 2 and 4 (TLR2 and TLR4) agonists differentially regulate secretory interleukin-1 receptor antagonist gene expression in macrophages. AB - Treatment of macrophages with lipopolysaccharide (LPS) from Gram-negative bacteria or peptidoglycan (PGN) from Gram-positive bacteria activates multiple intracellular signaling pathways and a large, diverse group of nuclear transcription factors. The signaling receptors for PGN and LPS are now known to be the Toll-like receptors 2 and 4 (TLR2 and -4, respectively). While a large body of literature indicates that the members of the TLR family activate nearly identical cytoplasmic signaling programs, several recent reports have suggested that the functional outcomes of signaling via TLR2 or TLR4 are not equivalent. In the current studies, we compared the responses of the secretory IL-1 receptor antagonist (sIL-1Ra) gene to both LPS and PGN. Both LPS and PGN induced IL-1Ra gene expression; however, the combination of both stimuli synergistically increased sIL-1Ra mRNA expression and promoter activity, suggesting that the signals induced by PGN and LPS are not equivalent. While both LPS and PGN utilized the PU.1-binding sites in the proximal sIL-1Ra promoter region to generate a full response, additional distinct promoter elements were utilized by LPS or PGN. Activation of p38 stress-activated protein kinase was required for LPS- or PGN-induced IL-1Ra gene expression, but the p38-responsive promoter elements localized to distinct regions of the sIL-1Ra gene. Additionally, while the LPS-induced, p38-dependent response was dependent upon PU.1 binding, the PGN induced, p38 response was not. Collectively, these data indicated that while some of the intracellular signaling events by TLR2 and TLR4 agonists are similar, there are clearly distinct differences in the responses elicited by these two bacterial products. PMID- 11877430 TI - Interaction between tyrosine kinase Etk and a RUN domain- and FYVE domain containing protein RUFY1. A possible role of ETK in regulation of vesicle trafficking. AB - Etk/BMX tyrosine kinase is involved in regulation of various cellular processes including proliferation, differentiation, motility, and apoptosis. Through a yeast two-hybrid screening for the effectors of Etk, a new gene family designated as RUFY was identified. The RUFY gene family (RUFY1 and RUFY2) contains an N terminal RUN domain and a C-terminal FYVE domain with two coiled-coil domains in between. They appear to be homologues of a recently identified mouse Rabip4 (Cormant, M., Mari, M., Galmiche, A., Hofman, P., and Le Marchand-Brustel, Y. (2001) Proc. Natl. Acad. Sci. U. S. A. 98, 1637-1642). RUFY proteins are localized predominantly to endosomes as evidenced by their co-localization with early endosome antigen marker (EEA1). Etk interacts with RUFY1 through its SH3 and SH2 domains. RUFY1 is tyrosine-phosphorylated and appears to be a substrate of Etk. The RUFY1 mutant lacking the phosphorylation sites failed to go to the endosomes. Furthermore, overexpression of Etk in COS-1 and B82L cells resulted in increased plasma membrane localization of the epidermal growth factor receptor and delayed its induced endocytosis in COS-1 cells. The effects of Etk were blocked by the FYVE domain of RUFY1. Interestingly, the FYVE domain of RUFY1 is targeted to the plasma membrane through an interaction between its proline-rich motif and the SH3 domain of Etk or possibly some other membrane-associated SH3 domain-containing protein(s), whereas the lipid binding activity of the FYVE domain is not required. Our data suggest that Etk may be involved in regulation of endocytosis through its interaction with an endosomal protein RUFY1. PMID- 11877431 TI - Stimulation of phospholipase C-epsilon by the M3 muscarinic acetylcholine receptor mediated by cyclic AMP and the GTPase Rap2B. AB - Stimulation of phospholipase C (PLC) by G(q)-coupled receptors such as the M(3) muscarinic acetylcholine receptor (mAChR) is caused by direct activation of PLC beta enzymes by Galpha(q) proteins. We have recently shown that G(s)-coupled receptors can stimulate PLC-epsilon, apparently via formation of cyclic AMP and activation of the Ras-related GTPase Rap2B. Here we report that PLC stimulation by the M(3) mAChR expressed in HEK-293 cells also involves, in part, similar mechanisms. M(3) mAChR-mediated PLC stimulation and [Ca(2+)](i) increase were reduced by 2',5'-dideoxyadenosine (dd-Ado), a direct adenylyl cyclase inhibitor. On the other hand, overexpression of Galpha(s) or Epac1, a cyclic AMP-regulated guanine nucleotide exchange factor for Rap GTPases, enhanced M(3) mAChR-mediated PLC stimulation. Inactivation of Ras-related GTPases with clostridial toxins suppressed the M(3) mAChR responses. The inhibitory toxin effects were mimicked by expression of inactive Rap2B, but not of other inactive GTPases (Rac1, Ras, RalA, Rap1A, and Rap2A). Activation of the M(3) mAChR induced GTP loading of Rap2B, an effect strongly enhanced by overexpression of Galpha(s) and inhibited by dd-Ado. Overexpression of PLC-epsilon and PLC-beta1, but not PLC-gamma1 or PLC delta1, enhanced M(3) mAChR-mediated PLC stimulation and [Ca(2+)](i) increase. In contrast, expression of a catalytically inactive PLC-epsilon mutant reduced PLC stimulation by the M(3) mAChR and abrogated the potentiating effect of Galpha(s). In conclusion, our findings suggest that PLC stimulation by the M(3) mAChR is a composite action of PLC-beta1 stimulation by Galpha(q) and stimulation of PLC epsilon apparently mediated by G(s)-dependent cyclic AMP formation and subsequent activation of Rap2B. PMID- 11877432 TI - Crystal structure of Thermotoga maritima 0065, a member of the IclR transcriptional factor family. AB - Members of the IclR family of transcription regulators modulate signal-dependent expression of genes involved in carbon metabolism in bacteria and archaea. The Thermotoga maritima TM0065 gene codes for a protein (TM-IclR) that is homologous to the IclR family. We have determined the crystal structure of TM-IclR at 2.2 A resolution using MAD phasing and synchrotron radiation. The protein is composed of two domains: the N-terminal DNA-binding domain contains the winged helix-turn helix motif, and the C-terminal presumed regulatory domain is involved in binding signal molecule. In a proposed signal-binding site, a bound Zn(2+) ion was found. In the crystal, TM-IclR forms a dimer through interactions between DNA-binding domains. In the dimer, the DNA-binding domains are 2-fold related, but the dimer is asymmetric with respect to the orientation of signal-binding domains. Crystal packing analysis showed that TM-IclR dimers form a tetramer through interactions exclusively by signal-binding domains. A model is proposed for binding of IclR like factors to DNA, and it suggests that signal-dependent transcription regulation is accomplished by affecting an oligomerization state of IclR and therefore its affinity for DNA target. PMID- 11877433 TI - Direct and novel regulation of cAMP-dependent protein kinase by Mck1p, a yeast glycogen synthase kinase-3. AB - The MCK1 gene of Saccharomyces cerevisiae encodes a protein kinase homologous to metazoan glycogen synthase kinase-3. Previous studies implicated Mck1p in negative regulation of pyruvate kinase. In this study we find that purified Mck1p does not phosphorylate pyruvate kinase, suggesting that the link is indirect. We find that purified Tpk1p, a cAMP-dependent protein kinase catalytic subunit, phosphorylates purified pyruvate kinase in vitro, and that loss of the cAMP dependent protein kinase regulatory subunit, Bcy1p, increases pyruvate kinase activity in vivo. We find that purified Mck1p inhibits purified Tpk1p in vitro, in the presence or absence of Bcy1p. Mck1p must be catalytically active to inhibit Tpk1p, but Mck1p does not phosphorylate this target. We find that abolition of Mck1p autophosphorylation on tyrosine prevents the kinase from efficiently phosphorylating exogenous substrates, but does not block its ability to inhibit Tpk1p in vitro. We find that this mutant form of Mck1p appears to retain the ability to negatively regulate cAMP-dependent protein kinase in vivo. We propose that Mck1p, in addition to phosphorylating some target proteins, also acts by a separate, novel mechanism: autophosphorylated Mck1p binds to and directly inhibits, but does not phosphorylate, the catalytic subunits of cAMP dependent protein kinase. PMID- 11877434 TI - Equilibrium binding assays reveal the elevated stoichiometry and salt dependence of the interaction between full-length human sex-determining region on the Y chromosome (SRY) and DNA. AB - In an effort to better define the molecular mechanism of the functional specificity of human sex-determining region on the Y chromosome (SRY), we have carried out equilibrium binding assays to study the interaction of the full length bacterial-expressed protein with a DNA response element derived from the CD3epsilon gene enhancer. These assays are based on the observation of the fluorescence anisotropy of a fluorescein moiety covalently bound to the target oligonucleotide. The low anisotropy value due to the fast tumbling of the free oligonucleotide in solution increases substantially upon binding the protein to the labeled target DNA. Our results indicate that the full-length human wild-type SRY (SRY(WT)) forms a complex of high stoichiometry with its target DNA. Moreover, we have demonstrated a strong salt dependence of both the affinity and specificity of the interaction. We have also addressed the DNA bending properties of full-length human SRY(WT) in solution by fluorescence resonance energy transfer and revealed that maximal bending is achieved with a protein to DNA ratio significantly higher than the classical 1:1. Oligomerization thus appears, at least in vitro, to be tightly coupled to SRY-DNA interactions. Alteration of protein-protein interactions observed for the mutant protein SRY(Y129N), identified in a patient presenting with 46,XY sex reversal, suggests that oligomerization may play an important role in vivo as well. PMID- 11877435 TI - Cellular arachidonate-releasing function and inflammation-associated expression of group IIF secretory phospholipase A2. AB - Here we report the cellular arachidonate (AA)-releasing function of group IIF secretory phospholipase A(2) (sPLA(2)-IIF), a sPLA(2) enzyme uniquely containing a longer C-terminal extension. sPLA(2)-IIF increased spontaneous and stimulus dependent release of AA, which was supplied to downstream cyclooxygenases and 5 lipoxygenase for eicosanoid production. sPLA(2)-IIF also enhanced interleukin 1 stimulated expression of cyclooxygenase-2 and microsomal prostaglandin E synthase. AA release by sPLA(2)-IIF was facilitated by oxidative modification of cellular membranes. Cellular actions of sPLA(2)-IIF occurred independently of the heparan sulfate proteoglycan glypican, which acts as a functional adaptor for other group II subfamily sPLA(2)s. Confocal microscopy revealed the location of sPLA(2)-IIF on the plasma membrane. The unique C-terminal extension was crucial for its plasma membrane localization and optimal cellular functions. sPLA(2)-IIF expression was increased in various tissues from lipopolysaccharide-treated mice and in ears of mice with experimental atopic dermatitis. In human rheumatoid arthritic joints, sPLA(2)-IIF was detected in synovial lining cells, capillary endothelial cells, and plasma cells. These results suggest that sPLA(2)-IIF is a potent regulator of AA metabolism and participates in the inflammatory process under certain conditions. PMID- 11877436 TI - Role of p21 in apoptosis and senescence of human colon cancer cells treated with camptothecin. AB - Treatment of cells with the anti-cancer drug camptothecin (CPT) induces topoisomerase I (Top1)-mediated DNA damage, which in turn affects cell proliferation and survival. In this report, we demonstrate that treatment of the wild-type HCT116 (wt HCT116) human colon cancer cell line and the isogenic p53(-/ ) HCT116 and p21(-/-) HCT116 cell lines with a high concentration (250 nm) of CPT resulted in apoptosis, indicating that apoptosis occurred by a p53- and p21 independent mechanism. In contrast, treatment with a low concentration (20 nm) of CPT induced cell cycle arrest and senescence of the wt HCT116 cells, but apoptosis of the p53(-/-) HCT116 and p21(-/-) HCT116 cells. Further investigations indicated that p53-dependent expression of p21 blocked apoptosis of wt HCT116 cells treated with 20 nm, but not 250 nm CPT. Interestingly, blocking of the apoptotic pathway, by Z-VAD-FMK, in p21(-/-) HCT116 cells following treatment with 20 nm CPT did not permit the cells to develop properties of senescence. These observations demonstrated that p21 was required for senescence development of HCT116 cells following treatment with low concentrations of CPT. PMID- 11877438 TI - A differentially methylated imprinting control region within the Kcnq1 locus harbors a methylation-sensitive chromatin insulator. AB - The mechanisms underlying the phenomenon of genomic imprinting remain poorly understood. In one instance, a differentially methylated imprinting control region (ICR) at the H19 locus has been shown to involve a methylation-sensitive chromatin insulator function that apparently partitions the neighboring Igf2 and H19 genes in different expression domains in a parent of origin-dependent manner. It is not known, however, if this mechanism is unique to the Igf2/H19 locus or if insulator function is a common feature in the regulation of imprinted genes. To address this question, we have studied an ICR in the Kcnq1 locus that regulates long range repression on the paternally derived p57Kip2 and Kcnq1 alleles in an imprinting domain that includes Igf2 and H19. We show that this ICR appears to possess a unidirectional chromatin insulator function in somatic cells of both mesodermal and endodermal origins. Moreover, we document that CpG methylation regulates this insulator function suggesting that a methylation-sensitive chromatin insulator is a common theme in the phenomenon of genomic imprinting. PMID- 11877437 TI - A role for the Psi-U mismatch in the recognition of the 5' splice site of yeast introns by the U1 small nuclear ribonucleoprotein particle. AB - The U1 small nuclear ribonucleoprotein particle (snRNP)/5' splice site (5'SS) interaction in yeast is essential for the splicing process and depends on the formation of a short RNA duplex between the 5' arm of U1 snRNA and the 1st intronic nucleotides. This RNA/RNA interaction is characterized by the presence of a mismatch that occurs with almost all yeast introns and concerns nucleotides 4 on the pre-mRNA (a U) and 5 on U1 snRNA (a Psi). The latter nucleotide is well conserved from yeast to vertebrates, but its role in yeast and the significance of the associated mismatch in the U1 snRNA/5'SS interaction have never been fully explained. We report here that the presence of this mismatch is a determinant of stability that mainly affects the off rate of the interaction. To our knowledge this is the first report assigning a function to this noncanonical interaction. We also performed SELEX (systematic evolution of ligands by exponential enrichment) experiments by immunoprecipitating U1 snRNP and the associated RNA. The artificial phylogeny derived from these experiments allows the isolation of the selective pressure due to U1 snRNP binding on the 5'SS of yeast introns. PMID- 11877439 TI - Identification of dynein heavy chain 7 as an inner arm component of human cilia that is synthesized but not assembled in a case of primary ciliary dyskinesia. AB - Although the basic structure of the axoneme has been highly conserved throughout evolution, the varied functions of specialized axonemes require differences in structure and regulation. Cilia lining the respiratory tract propel mucus along airway surfaces, providing a critical function to the defense mechanisms of the pulmonary system, yet little is known of their molecular structure. We have identified and cloned a dynein heavy chain that is a component of the inner dynein arm. Bronchial epithelial cells were obtained from normal donors and from a patient with primary ciliary dyskinesia (PCD) whose cilia demonstrated an absence of inner dynein arms by electron microscopy. Cilia from normal and PCD cells were compared by gel electrophoresis, and mass spectrometry was used to identify DNAH7 as a protein absent in PCD cilia. The full-length DNAH7 cDNA was cloned and shares 68% similarity with an inner arm dynein heavy chain from Drosophila. DNAH7 was induced during ciliated cell differentiation, and immunohistochemistry demonstrated the presence of DNAH7 in normal cilia. In cilia from PCD cells, DNAH7 was undetectable, whereas intracellular DNAH7 was clearly present. These studies identify DNAH7 as an inner arm component of human cilia that is synthesized but not assembled in a case of PCD. PMID- 11877440 TI - Protein kinase C delta regulates function of the DF3/MUC1 carcinoma antigen in beta-catenin signaling. AB - The DF3/MUC1 mucin-like glycoprotein is aberrantly overexpressed in most human carcinomas. The MUC1 cytoplasmic domain interacts directly with beta-catenin, a component of the adherens junction of mammalian epithelial cells. The present results demonstrate that MUC1 associates with protein kinase Cdelta (PKCdelta). A TDR sequence adjacent to the beta-catenin binding motif in the MUC1 cytoplasmic domain functions as a site for PKCdelta phosphorylation. We show that phosphorylation of MUC1 by PKCdelta increases binding of MUC1 and beta-catenin in vitro and in vivo. The functional significance of the MUC1-PKCdelta interaction is further supported by the demonstration that mutation of the PKCdelta phosphorylation site abrogates MUC1-mediated decreases in binding of beta-catenin to E-cadherin. We also show that the stimulatory effects of MUC1 on anchorage independent growth are abrogated by mutation of the PKCdelta phosphorylation site. These findings support a novel role for PKCdelta in regulating the interaction between MUC1 and the beta-catenin signaling pathway. PMID- 11877441 TI - Transcriptional regulation of cyclooxygenase-1 by histone deacetylase inhibitors in normal human astrocyte cells. AB - While cyclooxygenase (COX)-2 is a highly inducible gene, COX-1 is widely known as a noninducible gene and is constitutively expressed in a variety of cell lines and human tissues. Recently, several reports have indicated that COX-1 is also regulated at the transcriptional level by various stimuli. We present evidence that histone deacetylase (HDAC) inhibitors induce COX-1 transcription and translation in normal human astrocyte (NHA) cells and glioma cell lines. HDAC inhibitors increased acetylated histone H4 protein expression in NHA cells. The levels of COX-1 mRNA and protein were maximal at 24 and 48 h, respectively, after treatment with the specific HDAC inhibitor, trichostatin A (TSA). In addition, TSA-treated NHA cells produced prostaglandin E(2) as determined by enzyme-linked immunosorbent assay after incubation with 10 microm exogenous arachidonic acid, indicating that the induced COX-1 is functionally active. In addition to NHA cells, this up-regulation of COX-1 after treatment with HDAC inhibitors was observed in 5 different glioma cell lines. The nucleotide sequence of the inducible COX-1 cDNA was confirmed identical to human COX-1 that was previously reported. HDAC inhibitors stimulated COX-1 promoter activity as measured by luciferase reporter assays, suggesting that the induction of COX-1 is regulated at the transcriptional level. Furthermore, mutation analysis of the COX-1 promoter suggests that TSA-responsive element exists in the proximal Sp1-binding site at +25 to +31. In conclusion, COX-1 is an inducible gene in glial-derived cells including immortalized cells, and appears to be transcriptionally regulated by a unique mechanism associated with histone acetylation. PMID- 11877442 TI - Thioredoxin-2 but not thioredoxin-1 is a substrate of thioredoxin peroxidase-1 from Drosophila melanogaster: isolation and characterization of a second thioredoxin in D. Melanogaster and evidence for distinct biological functions of Trx-1 and Trx-2. AB - As Drosophila melanogaster does not contain glutathione reductase, the thioredoxin system has a key function for glutathione disulfide reduction in insects (Kanzok, S. M., Fechner, A., Bauer, H., Ulschmid, J. K., Muller, H. M., Botella-Munoz, J., Schneuwly, S., Schirmer, R. H., and Becker, K. (2001) Science 291, 643-646). In view of these unique conditions, the protein systems participating in peroxide metabolism and in redox signaling are of special interest. The genes for a second thioredoxin (DmTrx-2) and a thioredoxin peroxidase (DmTPx-1) were cloned and expressed, and the proteins were characterized. In its disulfide form, the 13-kDa protein thioredoxin-2 is a substrate of thioredoxin reductase-1 (K(m) = 5.2 microm, k(cat) = 14.5 s(-1)) and in its dithiol form, an electron donor for TPx-1 (K(m) = 9 microm, k(cat) = 5.4 s(-1)). DmTrx-2 is capable of reducing glutathione disulfide with a second order rate constant of 170 m(-1) s(-1) at pH 7.4 and 25 degrees C. Western blot analysis indicated that this thioredoxin represents up to 1% of the extractable protein of D. melanogaster Schneider cells or whole fruit flies. Recombinant thioredoxin peroxidase-1 (subunit molecular mass = 23 kDa) was found to be a decameric protein that can efficiently use Trx-2 but not Trx-1 as a reducing substrate. The new electron pathway found in D. melanogaster is also representative for insects that serve as vectors of disease. As a first step we have cloned and functionally expressed the gene that is the orthologue of DmTrx-2 in the malaria mosquito Anopheles gambiae. PMID- 11877443 TI - Tn5 transposase active site mutants. AB - Tn5 transposase (Tnp) is a 53.3-kDa protein that is encoded by and facilitates movement of transposon Tn5. Tnp monomers contain a single active site that is responsible for catalyzing a series of four DNA breaking/joining reactions at one transposon end. Based on primary sequence homology and protein structural information, we designed and constructed a series of plasmids that encode for Tnps containing active site mutations. Following Tnp expression and purification, the active site mutants were tested for their ability to form protein-DNA complexes and perform each of the four catalytic steps in the transposition pathway in vitro. The results demonstrate that Asp-97, Asp-188, and Glu-326, visible in the active site of Tn5 crystal structures, are absolutely required for all catalytic steps. Mutations within a series of amino acid residues that are conserved in the IS4 family of transposases and retroviral integrases also impair Tnp catalytic activity. Mutations at either Tyr-319 or Arg-322 reduce both hairpin resolution and strand transfer activity within protein-DNA complexes. Mutations at Lys-333 reduce the ability of Tnps to form protein-DNA complexes, whereas mutations at the less strongly conserved Lys-330 have less of an effect on both synaptic complex formation and catalytic activity. PMID- 11877444 TI - T0070907, a selective ligand for peroxisome proliferator-activated receptor gamma, functions as an antagonist of biochemical and cellular activities. AB - The nuclear hormone receptor peroxisome proliferator-activated receptor gamma (PPARgamma (NR1C3)) plays a central role in adipogenesis and is the molecular target for the thiazolidinedione (TZD) class of antidiabetic drugs. In a search for novel non-TZD ligands for PPARgamma, T0070907 was identified as a potent and selective PPARgamma antagonist. With an apparent binding affinity (concentration at 50% inhibition of [(3)H]rosiglitazone binding or IC(50)) of 1 nm, T0070907 covalently modifies PPARgamma on cysteine 313 in helix 3 of human PPARgamma2. T0070907 blocked PPARgamma function in both cell-based reporter gene and adipocyte differentiation assays. Consistent with its role as an antagonist of PPARgamma, T0070907 blocked agonist-induced recruitment of coactivator-derived peptides to PPARgamma in a homogeneous time-resolved fluorescence-based assay and promoted recruitment of the transcriptional corepressor NCoR to PPARgamma in both glutathione S-transferase pull-down assays and a PPARgamma/retinoid X receptor (RXR) alpha-dependent gel shift assay. Studies with mutant receptors suggest that T0070907 modulates the interaction of PPARgamma with these cofactor proteins by affecting the conformation of helix 12 of the PPARgamma ligand-binding domain. Interestingly, whereas the T0070907-induced NCoR recruitment to PPARgamma/RXRalpha heterodimer can be almost completely reversed by the simultaneous treatment with RXRalpha agonist LGD1069, T0070907 treatment has only modest effects on LGD1069-induced coactivator recruitment to the PPARgamma/RXRalpha heterodimer. These results suggest that the activity of PPARgamma antagonists can be modulated by the availability and concentration of RXR agonists. T0070907 is a novel tool for the study of PPARgamma/RXRalpha heterodimer function. PMID- 11877445 TI - CXCR4/CCR5 down-modulation and chemotaxis are regulated by the proteasome pathway. AB - Chemokines and their receptors play a critical role in host immune surveillance and are important mediators of human immunodeficiency virus (HIV) pathogenesis and inflammatory response. The chemokine receptors CCR5 and CXCR4, which act as co-receptors along with CD4 for HIV docking and entry, are down-modulated by their respective ligands, MIP-1beta/SDF-1alpha or by the HIV envelope protein, gp120. We have studied the role of the proteasome pathway in the down-regulation of these receptors. Using the yeast and mammalian two-hybrid systems, we observed that the CCR5 receptor is constitutively associated with the zeta subunit of proteasome. Immunoprecipitation studies in CCR5 L1.2 cells revealed that this association was increased with MIP-1beta stimulation. The proteasome inhibitors, lactacystin and epoxomicin, attenuated MIP-1beta induced CCR5 down-modulation as detected by fluorescence-activated cell sorter analysis and confocal microscopy. The proteasome inhibitors also inhibited the SDF-1alpha and gp120 protein-induced down-modulation of the CXCR4 receptor in Jurkat cells. However, the inhibitors had no significant effect on the gp120-induced internalization of the CD4 receptor. These inhibitors also blocked cognate ligand-mediated chemotaxis but had no effect on SDF-1alpha-induced p44/42 MAP kinase or MIP-1beta-induced p38 kinase activities, thus indicating differential effects of the inhibitors on signaling mediated by these receptors. These results indicate that the CCR5 and CXCR4 receptor down-modulation mechanism and chemotaxis mediated by these receptors are dependent upon proteasome activity. PMID- 11877446 TI - Trypanosoma brucei FLA1 is required for flagellum attachment and cytokinesis. AB - The single flagellum of the protozoan parasite Trypanosoma brucei is attached along the length of the cell body by a complex structure that requires the FLA1 protein. We show here that inhibition of FLA1 expression by RNA interference in procyclic trypanosomes causes flagellar detachment and prevents cytokinesis. Despite being unable to divide, these cells undergo mitosis and develop a multinucleated phenotype. The Trypanosoma cruzi FLA1 homolog, GP72, is unable to complement either the flagellar detachment or cytokinesis defects in procyclic T. brucei that have been depleted of FLA1 by RNA interference. Instead, GP72 itself caused flagellar detachment when expressed in T. brucei. In contrast to T. brucei cells depleted of FLA1, procyclic T. brucei expressing GP72 continued to divide despite having detached flagella, demonstrating that flagellar attachment is not absolutely necessary for cytokinesis. We have also identified a FLA1-related gene (FLA2) whose sequence is similar but not identical to FLA1. Inhibition of FLA1 and FLA2 expression in bloodstream T. brucei caused flagellar detachment and blocked cytokinesis but did not inhibit mitosis. These experiments demonstrate that the FLA proteins are essential and suggest that in procyclic T. brucei, the FLA1 protein has separable functions in flagellar attachment and cytokinesis. PMID- 11877447 TI - Subcellular localization of Aft1 transcription factor responds to iron status in Saccharomyces cerevisiae. AB - The Aft1 transcription factor regulates the iron regulon in response to iron availability in Saccharomyces cerevisiae. Aft1 activates a battery of genes required for iron uptake under iron-starved conditions, whereas Aft1 function is inactivated under iron-replete conditions. Previously, we have shown that iron regulated DNA binding by Aft1 is responsible for the controlled expression of target genes. Here we show that this iron-regulated DNA binding by Aft1 is not due to the change in the total expression level of Aft1 or alteration of DNA binding activity. Rather, nuclear localization of Aft1 responds to iron status, leading to iron-regulated expression of the target genes. We identified the nuclear export signal (NES)-like sequence in the AFT1 open reading frame. Mutation of the NES-like sequence causes nuclear retention of Aft1 and the constitutive activation of Aft1 function independent of the iron status of the cells. These results suggest that the nuclear export of Aft1 is critical for ensuring iron-responsive transcriptional activation of the Aft1 regulon and that the nuclear import/export systems are involved in iron sensing by Aft1 in S. cerevisiae. PMID- 11877448 TI - Regulation of internal ribosomal entry site-mediated translation by phosphorylation of the translation initiation factor eIF2alpha. AB - Initiation of translation from most cellular mRNAs occurs via scanning; the 40 S ribosomal subunit binds to the m(7)G-cap and then moves along the mRNA until an initiation codon is encountered. Some cellular mRNAs contain internal ribosome entry sequences (IRESs) within their 5'-untranslated regions, which allow initiation independently of the 5'-cap. This study investigated the ability of cellular stress to regulate the activity of IRESs in cellular mRNAs. Three stresses were studied that cause the phosphorylation of the translation initiation factor, eIF2alpha, by activating specific kinases: (i) amino acid starvation, which activates GCN2; (ii) endoplasmic reticulum (ER) stress, which activates PKR-like ER kinase, PERK kinase; and (iii) double-stranded RNA, which activates double-stranded RNA-dependent protein kinase (PKR) by mimicking viral infection. Amino acid starvation and ER stress caused transient phosphorylation of eIF2alpha during the first hour of treatment, whereas double-stranded RNA caused a sustained phosphorylation of eIF2alpha after 2 h. The effects of these treatments on IRES-mediated initiation were investigated using bicistronic mRNA expression vectors. No effect was seen for the IRESs from the mRNAs for the chaperone BiP and the protein kinase Pim-1. In contrast, translation mediated by the IRESs from the cationic amino acid transporter, cat-1, and of the cricket paralysis virus intergenic region, were stimulated 3- to 10-fold by all three treatments. eIF2alpha phosphorylation was required for the response because inactivation of phosphorylation prevented the stimulation. It is concluded that cellular stress can stimulate translation from some cellular IRESs via a mechanism that requires the phosphorylation of eIF2alpha. Moreover, there are distinct regulatory patterns for different cellular mRNAs that contain IRESs within their 5'-untranslated regions. PMID- 11877449 TI - A novel type I cytokine receptor is expressed on monocytes, signals proliferation, and activates STAT-3 and STAT-5. AB - Here we report the cloning of a novel type I cytokine receptor, gp130-like monocyte receptor (GLM-R), with homology to the interleukin-6 receptor signal transducing chain, gp130, and granulocyte colony-stimulating factor receptor. Human and murine GLM-R cDNAs encode open reading frames of 732 and 716 amino acids, respectively, and the corresponding genes are located in close proximity to gp130 genes on human chromosome 5 and mouse chromosome 13. GLM-R is specifically expressed on CD14-positive cells and is up-regulated more than 50 fold upon activation of those cells. To address the question of whether GLM-R is a signaling receptor, we constructed a chimeric molecule, consisting of the extracellular domain of human growth hormone (hGH) receptor, and the intracellular domain of GLM-R. When transfected into factor-dependent 32D cells, this chimeric molecule could signal for proliferation and activate signal transducer and activator of transcription (STAT)-3 and STAT-5 upon stimulation with hGH. Thus, GLM-R is a novel signaling receptor chain potentially involved in the development and function of monocytes and macrophages. PMID- 11877450 TI - Kaurane diterpene, kamebakaurin, inhibits NF-kappa B by directly targeting the DNA-binding activity of p50 and blocks the expression of antiapoptotic NF-kappa B target genes. AB - Kaurane diterpenes have been identified from numerous medicinal plants, which have been used for treatment of inflammation and cancer, however, their molecular mechanism of action remains unclear. We have previously shown that kamebakaurin and other three kaurane diterpenes selectively inhibit activation of transcription factor NF-kappaB, a central mediator of apoptosis and immune responses. We here demonstrate that kamebakaurin is a potent inhibitor of NF kappaB activation by directly targeting DNA-binding activity of p50. Kamebakaurin prevented the activation of NF-kappaB by different stimuli in various cell types. Kamebakaurin did not prevent either stimuli-induced degradation of IkappaB-alpha or nuclear translocation of NF-kappaB, however, it significantly interfered DNA binding activity of activated NF-kappaB in cell and in vitro and preferentially prevented p50-mediated DNA-binding activity of NF-kappaB rather than that of RelA as measured using in vitro translated p50 and RelA proteins. Moreover, a p50 mutant with a Cys-62 --> Ser mutation was not inhibited with kamebakaurin, indicating that the effect of kamebakaurin was probably due to its interaction with cysteine 62 in p50. The covalent modification of p50 by kamebakaurin was further demonstrated by mass spectrometry analysis that showed an increase in the molecular mass of kamebakaurin-treated p50, and this modification was not reverted by addition of dithiothreitol. These results suggested that kamebakaurin exhibited its inhibitory activity by a direct covalent modification of cysteine 62 in the p50. Also, treatment of cells with kamebakaurin prevented the tumor necrosis factor-alpha (TNF-alpha)-induced expression of antiapoptotic NF-kappaB target genes encoding c-IAP1 (hiap-2) and c-IAP2 (hiap-1), members of the inhibitor of apoptosis family, and Bfl-1/A1, a prosurvival Bcl-2 homologue, and augmented the TNF-alpha-induced caspase 8 activity, thereby resulting in sensitizing MCF-7 cells to TNF-alpha-induced apoptosis. Taken together, kamebakaurin is a valuable candidate for the intervention of NF-kappaB-dependent pathological conditions such as inflammation and cancer. PMID- 11877451 TI - Regulation of arrestin-3 phosphorylation by casein kinase II. AB - Arrestins play an important role in regulating the function of G protein-coupled receptors including receptor desensitization, internalization, down-regulation, and signaling via nonreceptor tyrosine kinases and mitogen-activated protein kinases. Previous studies have revealed that arrestins themselves are also subject to regulation. In the present study, we focused on identifying potential mechanisms involved in regulating the function of arrestin-3. Using metabolic labeling, phosphoamino acid analysis, and mutagenesis studies, we found that arrestin-3 is constitutively phosphorylated at Thr-382 and becomes dephosphorylated upon beta(2)-adrenergic receptor activation in COS-1 cells. Casein kinase II (CKII) appears to be the major kinase mediating arrestin-3 phosphorylation, since 1) Thr-382 is contained within a canonical consensus sequence for CKII phosphorylation and 2) wild type arrestin-3 but not a T382A mutant is phosphorylated by CKII in vitro. Functional analysis reveals that mutants mimicking the phosphorylated (T382E) and dephosphorylated (T382A or T382V) states of arrestin-3 promote beta(2)-adrenergic receptor internalization and bind clathrin, beta-adaptin, and Src to comparable levels as wild type arrestin-3. This suggests that the phosphorylation of arrestin-3 does not directly regulate interaction with endocytic (clathrin, beta-adaptin) or signaling (Src) components and is in contrast to arrestin-2, where phosphorylation appears to regulate interaction with clathrin and Src. However, additional analysis reveals that arrestin-3 phosphorylation may regulate formation of a large arrestin-3-containing protein complex. Differences between the regulatory roles of arrestin-2 and -3 phosphorylation may contribute to the different cellular functions of these proteins in G protein-coupled receptor signaling and regulation. PMID- 11877452 TI - Increased K+ efflux and apoptosis induced by the potassium channel modulatory protein KChAP/PIAS3beta in prostate cancer cells. AB - K(+) channel-associated protein/protein inhibitor of activated STAT (KChAP/PIAS3beta) is a potassium (K(+)) channel modulatory protein that boosts protein expression of a subset of K(+) channels and increases currents without affecting gating. Since increased K(+) efflux is an early event in apoptosis, we speculated that KChAP might induce apoptosis through its up-regulation of K(+) channel expression. KChAP belongs to the protein inhibitor of activated STAT family, members of which also interact with a variety of transcription factors including the proapoptotic protein, p53. Here we report that KChAP induces apoptosis in the prostate cancer cell line, LNCaP, which expresses both K(+) currents and wild-type p53. Infection with a recombinant adenovirus encoding KChAP (Ad/KChAP) increases K(+) efflux and reduces cell size as expected for an apoptotic volume decrease. The apoptosis inducer, staurosporine, increases endogenous KChAP levels, and LNCaP cells, 2 days after Ad/KChAP infection, show increased sensitivity to staurosporine. KChAP increases p53 levels and stimulates phosphorylation of p53 residue serine 15. Consistent with activation of p53 as a transcription factor, p21 levels are increased in infected cells. Wild-type p53 is not essential for induction of apoptosis by KChAP, however, since KChAP also induces apoptosis in DU145 cells, a prostate cancer cell line with mutant p53. Consistent with its proapoptotic properties, KChAP prevents growth of DU145 and LNCaP tumor xenografts in nude mice, indicating that infection with Ad/KChAP might represent a novel method of cancer treatment. PMID- 11877453 TI - NEMO trimerizes through its coiled-coil C-terminal domain. AB - NEMO/IkappaB kinase (IKK) gamma is the regulatory component of the IKK complex comprising the two protein kinases, IKKalpha and IKKbeta. To investigate the self assembly properties of NEMO and to understand further the mechanism of activation of the IKK complex, we purified wild-type and mutant NEMO expressed in Escherichia coli. In the absence of its IKK partners, recombinant NEMO (rNEMO) is a metastable functional monomer correctly folded, according to its fluorescence and far-UV CD spectra, which is binding specifically to the IKK complex. A minor fraction of rNEMO was found tightly associated with DnaK (E. coli Hsp70). We also examined the interaction of NEMO with prokaryotic and eukaryotic Hsp70, and we showed that the Hsp70-NEMO complex forms a supramolecular structure probably corresponding to an assembly intermediate. In vivo cross-linking experiments indicate that native NEMO in association with IKK is in equilibrium between a dimeric and a trimeric form. Similarly to native NEMO, a NEMO mutant deleted from its IKK binding N-terminal domain (residues 242-388) forms a stable trimeric coiled-coil, suggesting that the association of NEMO with IKK or with Hsp70 prevents incorrect interdomain pairing reactions that could lead to aggregation or to an non-native oligomeric state of rNEMO. We propose a model in which the activation of the IKK complex occurs through the trimerization of NEMO upon binding to a not yet identified upstream activator. PMID- 11877454 TI - NFAT: ubiquitous regulator of cell differentiation and adaptation. AB - The nuclear factor of activated T cells (NFAT) proteins are a family of transcription factors whose activation is controlled by calcineurin, a Ca(2+) dependent phosphatase. Originally identified in T cells as inducers of cytokine gene expression, NFAT proteins play varied roles in cells outside of the immune system. This review addresses the recent data implicating NFAT in the control of gene expression influencing the development and adaptation of numerous mammalian cell types. PMID- 11877455 TI - In vivo binding of active heat shock transcription factor 1 to human chromosome 9 heterochromatin during stress. AB - Activation of the mammalian heat shock transcription factor (HSF)1 by stress is a multistep process resulting in the transcription of heat shock genes. Coincident with these events is the rapid and reversible redistribution of HSF1 to discrete nuclear structures termed HSF1 granules, whose function is still unknown. Key features are that the number of granules correlates with cell ploidy, suggesting the existence of a chromosomal target. Here we show that in humans, HSF1 granules localize to the 9q11-q12 heterochromatic region. Within this locus, HSF1 binds through direct DNA-protein interaction with a nucleosome-containing subclass of satellite III repeats. HSF1 granule formation only requires the DNA binding competence and the trimerization of the factor. This is the first example of a transcriptional activator that accumulates transiently and reversibly on a chromosome-specific heterochromatic locus. PMID- 11877457 TI - Phosphorylation of the AP2 mu subunit by AAK1 mediates high affinity binding to membrane protein sorting signals. AB - During receptor-mediated endocytosis, AP2 complexes act as a bridge between the cargo membrane proteins and the clathrin coat by binding to sorting signals via the mu 2 subunit and to clathrin via the beta subunit. Here we show that binding of AP2 to sorting signals in vitro is regulated by phosphorylation of the mu 2 subunit of AP2. Phosphorylation of mu 2 enhances the binding affinity of AP2 for sorting motifs as much as 25-fold compared with dephosphorylated AP2. The recognition of sorting signals was not affected by the phosphorylation status of the alpha or beta 2 subunit, suggesting that phosphorylation of mu 2 is critical for regulation of AP2 binding to sorting signals. Phosphorylation of mu 2 occurs at a single threonine residue (Thr-156) and is mediated by the newly discovered adaptor-associated kinase, AAK1, which copurifies with AP2. We propose that phosphorylation of the AP2 mu 2 subunit by AAK1 ensures high affinity binding of AP2 to sorting signals of cargo membrane proteins during the initial steps of receptor-mediated endocytosis. PMID- 11877456 TI - CHO1, a mammalian kinesin-like protein, interacts with F-actin and is involved in the terminal phase of cytokinesis. AB - CHO1 is a kinesin-like protein of the mitotic kinesin-like protein (MKLP)1 subfamily present in central spindles and midbodies in mammalian cells. It is different from other subfamily members in that it contains an extra approximately 300 bp in the COOH-terminal tail. Analysis of the chicken genomic sequence showed that heterogeneity is derived from alternative splicing, and exon 18 is expressed in only the CHO1 isoform. CHO1 and its truncated isoform MKLP1 are coexpressed in a single cell. Surprisingly, the sequence encoded by exon 18 possesses a capability to interact with F-actin, suggesting that CHO1 can associate with both microtubule and actin cytoskeletons. Microinjection of exon 18-specific antibodies did not result in any inhibitory effects on karyokinesis and early stages of cytokinesis. However, almost completely separated daughter cells became reunited to form a binulceate cell, suggesting that the exon 18 protein may not have a role in the formation and ingression of the contractile ring in the cortex. Rather, it might be involved directly or indirectly in the membrane events necessary for completion of the terminal phase of cytokinesis. PMID- 11877458 TI - Transferrin receptor recycling in the absence of perinuclear recycling endosomes. AB - In mammalian cells, internalized receptors such as transferrin (Tfn) receptor are presumed to pass sequentially through early endosomes (EEs) and perinuclear recycling endosomes (REs) before returning to the plasma membrane. Whether passage through RE is obligatory, however, remains unclear. Kinetic analysis of endocytosis in CHO cells suggested that the majority of internalized Tfn bypassed REs returning to the surface from EEs. To determine directly if REs are dispensable for recycling, we studied Tfn recycling in cytoplasts microsurgically created to contain peripheral EEs but to exclude perinuclear REs. The cytoplasts actively internalized and recycled Tfn. Surprisingly, they also exhibited spatially and temporally distinct endosome populations. The first appeared to correspond to EEs, labeling initially with Tfn, being positive for early endosomal antigen 1 (EEA-1) and containing only small amounts of Rab11, an RE marker. The second was EEA-1 negative and with time recruited Rab11, suggesting that cytoplasts assembled functional REs. These results suggest that although perinuclear REs are not essential components of the Tfn recycling pathway, they are dynamic structures which preexist in the peripheral cytoplasm or can be regenerated from EE- and cytosol-derived components such as Rab11. PMID- 11877459 TI - The role of cell cycle-regulated expression in the localization of spatial landmark proteins in yeast. AB - In Saccharomyces cerevisiae, Bud8p and Bud9p are homologous plasma membrane glycoproteins that appear to mark the distal and proximal cell poles, respectively, as potential sites for budding in the bipolar pattern. Here we provide evidence that Bud8p is delivered to the presumptive bud site (and thence to the distal pole of the bud) just before bud emergence, and that Bud9p is delivered to the bud side of the mother-bud neck (and thence to the proximal pole of the daughter cell) after activation of the mitotic exit network, just before cytokinesis. Like the delivery of Bud8p, that of Bud9p is actin dependent; unlike the delivery of Bud8p, that of Bud9p is also septin dependent. Interestingly, although the transcription of BUD8 and BUD9 appears to be cell cycle regulated, the abundance of BUD8 mRNA peaks in G2/M and that of BUD9 mRNA peaks in late G1, suggesting that the translation and/or delivery to the cell surface of each protein is delayed and presumably also cell cycle regulated. The importance of time of transcription in localization is supported by promoter-swap experiments: expression of Bud8p from the BUD9 promoter leads to its localization predominantly to the sites typical for Bud9p, and vice versa. Moreover, expression of Bud8p from the BUD9 promoter fails to rescue the budding-pattern defect of a bud8 mutant but fully rescues that of a bud9 mutant. However, although expression of Bud9p from the BUD8 promoter fails to rescue a bud9 mutant, it also rescues only partially the budding-pattern defect of a bud8 mutant, suggesting that some feature(s) of the Bud8p protein is also important for Bud8p function. Experiments with chimeric proteins suggest that the critical element(s) is somewhere in the extracytoplasmic domain of Bud8p. PMID- 11877461 TI - Identification of an adaptor-associated kinase, AAK1, as a regulator of clathrin mediated endocytosis. AB - The mu 2 subunit of the AP2 complex is known to be phosphorylated in vitro by a copurifying kinase, and it has been demonstrated recently that mu 2 phosphorylation is required for transferrin endocytosis (Olusanya, O., P.D. Andrews, J.R. Swedlow, and E. Smythe. 2001. Curr. Biol. 11:896-900). However, the identity of the endogenous kinase responsible for this phosphorylation is unknown. Here we identify and characterize a novel member of the Prk/Ark family of serine/threonine kinases, adaptor-associated kinase (AAK)1. We find that AAK1 copurifies with adaptor protein (AP)2 and that it directly binds the ear domain of alpha-adaptin in vivo and in vitro. In neuronal cells, AAK1 is enriched at presynaptic terminals, whereas in nonneuronal cells it colocalizes with clathrin and AP2 in clathrin-coated pits and at the leading edge of migrating cells. AAK1 specifically phosphorylates the mu subunit in vitro, and stage-specific assays for endocytosis show that mu phosphorylation by AAK1 results in a decrease in AP2 stimulated transferrin internalization. Together, these results provide strong evidence that AAK1 is the endogenous mu 2 kinase and plays a regulatory role in clathrin-mediated endocytosis. These results also lend support to the idea that clathrin-mediated endocytosis is controlled by cycles of phosphorylation/desphosphorylation. PMID- 11877460 TI - High RhoA activity maintains the undifferentiated mesenchymal cell phenotype, whereas RhoA down-regulation by laminin-2 induces smooth muscle myogenesis. AB - Round embryonic mesenchymal cells have the potential to differentiate into smooth muscle (SM) cells upon spreading/elongation (Yang, Y., K.C. Palmer, N. Relan, C. Diglio, and L. Schuger. 1998. Development. 125:2621-2629; Yang, Y., N.K. Relan, D.A. Przywara, and L. Schuger. 1999. Development. 126:3027-3033; Yang, Y., S. Beqaj, P. Kemp, I. Ariel, and L. Schuger. 2000. J. Clin. Invest. 106:1321-1330). In the developing lung, this process is stimulated by peribronchial accumulation of laminin (LN)-2 (Relan, N.K., Y. Yang, S. Beqaj, J.H. Miner, and L. Schuger. 1999. J. Cell Biol. 147:1341-1350). Here we show that LN-2 stimulates bronchial myogenesis by down-regulating RhoA activity. Immunohistochemistry, immunoblotting, and reverse transcriptase-PCR indicated that RhoA, a small GTPase signaling protein, is abundant in undifferentiated embryonic mesenchymal cells and that its levels decrease along with SM myogenesis. Functional studies using agonists and antagonists of RhoA activation and dominant positive and negative plasmid constructs demonstrated that high RhoA activity was required to maintain the round undifferentiated mesenchymal cell phenotype. This was in part achieved by restricting the localization of the myogenic transcription factor serum response factor (SRF) mostly to the mesenchymal cell cytoplasm. Upon spreading on LN-2 but not on other main components of the extracellular matrix, the activity and level of RhoA decreased rapidly, resulting in translocation of SRF to the nucleus. Both cell elongation and SRF translocation were prevented by overexpression of dominant positive RhoA. Once the cells underwent SM differentiation, up-regulation of RhoA activity induced rather than inhibited SM gene expression. Therefore, our studies suggest a novel mechanism whereby LN-2 and RhoA modulate SM myogenesis. PMID- 11877462 TI - Intrinsic and innate defenses in the lung: intersection of pathways regulating lung morphogenesis, host defense, and repair. PMID- 11877464 TI - Apoptosis and NF-kappa B: the FADD connection. PMID- 11877463 TI - Mucus clearance as a primary innate defense mechanism for mammalian airways. PMID- 11877465 TI - Degeneracy, as opposed to specificity, in immunotherapy. PMID- 11877466 TI - Overexpression of the serpin megsin induces progressive mesangial cell proliferation and expansion. AB - Mesangial cells maintain normal glomerular function by mediating ECM remodeling and immune complex disposal. We have recently identified megsin, a novel member of the serine protease inhibitor (serpin) superfamily predominantly expressed in the mesangium. While our previous studies suggested a role for megsin in the pathogenesis of human glomerular diseases, its exact biological significance remained unknown. Here we produced two lines of megsin transgenic mice. Overexpression of megsin led to progressive mesangial matrix expansion and an increase in the number of mesangial cells. These glomerular lesions were accompanied by an augmented immune complex deposition, together with Ig's and complement. Binding and functional assays in vitro identified plasmin as one biological substrate of megsin and confirmed its activity as a proteinase inhibitor. Transgenic animals exhibiting nephritis as a result of treatment with anti--glomerular basement membrane antiserum showed significantly more persistent expansion of the mesangial ECM than was seen in parental mice. Megsin therefore exerts a biologically relevant influence on mesangial function, and on the mesangial microenvironment, such that simple overexpression of this endogenous serpin engenders elementary mesangial lesions. PMID- 11877467 TI - Suppression of body fat accumulation in myostatin-deficient mice. AB - Myostatin is a TGF-beta family member that acts as a negative regulator of muscle growth. Mice lacking the myostatin gene (Mstn) have a widespread increase in skeletal muscle mass resulting from a combination of muscle fiber hypertrophy and hyperplasia. Here we show that Mstn-null mice have a significant reduction in fat accumulation with increasing age compared with wild-type littermates, even in the setting of normal food intake (relative to body weight), normal body temperature, and a slightly decreased resting metabolic rate. To investigate whether myostatin might be an effective target for suppressing the development of obesity in settings of abnormal fat accumulation, we analyzed the effect of the Mstn mutation in two genetic models of obesity, agouti lethal yellow (A(y)) and obese (Lep(ob/ob)). In each case, loss of Mstn led to a partial suppression of fat accumulation and of abnormal glucose metabolism. Our findings raise the possibility that pharmacological agents that block myostatin function may be useful not only for enhancing muscle growth, but also for slowing or preventing the development of obesity and type 2 diabetes. PMID- 11877468 TI - Evidence for the importance of angiotensin II type 1 receptor in ischemia-induced angiogenesis. AB - The role of the renin-angiotensin system (RAS) in angiogenesis is little known. Here, we show that the angiotensin II (ATII) type 1 (AT1) receptor plays an important role in ischemia-induced angiogenesis. Well-developed collateral vessels and angiogenesis were observed in wild-type (WT) mice in response to hindlimb ischemia, whereas these responses were reduced in ATII type 1a receptor knockout (AT1a(-/-)) mice. Ischemia-induced angiogenesis was also impaired in WT mice treated with the AT1 receptor blocker TCV-116. These effects were not due to reduced systemic blood pressure (SBP), because hydralazine treatment preserved angiogenesis in WT mice although it reduced SBP to a level similar to that of AT1a(-/-) mice. Infiltration of inflammatory mononuclear cells (MNCs), including macrophages and T lymphocytes, was suppressed in the ischemic tissues of AT1a(-/ ) mice compared with WT mice. Double immunofluorescence staining revealed that infiltrated macrophages and T lymphocytes expressed VEGF, and the expression of VEGF and monocyte chemoattractant protein-1 was also decreased in AT1a(-/-). Finally, the impaired angiogenesis in AT1a(-/-) mice was rescued by intramuscular transplantation of MNCs obtained from WT mice, further indicating the importance of MNC infiltration in ischemia-induced angiogenesis. Thus, the ATII--AT1 receptor pathway promotes early angiogenesis by supporting inflammatory cell infiltration and angiogenic cytokine expression. PMID- 11877469 TI - Short-circuiting long-lived humoral immunity by the heightened engagement of CD40. AB - Agonistic alpha CD40 Ab's have been shown to be potent immune adjuvants for both cell- and humoral-mediated immunity. While enhancing short-lived humoral immunity, the administration of a CD40 agonist during thymus-dependent immune responses ablates germinal center formation, prematurely terminates the humoral immune response, blocks the generation of B cell memory, and prevents the generation of long-lived bone marrow plasma cells. Interestingly, some of these effects of heightened CD40 engagement could be mimicked by enhancing the magnitude of antigen-specific T cell help. Taken together, these studies demonstrate that as the magnitude of CD40 signaling intensifies, the fate of antigen-reactive B cells can be dramatically altered. These are the first studies to describe the multifaceted function of CD40 in determining the fate of antigen reactive B cells and provide novel insights into how CD40 agonists can short circuit humoral immunity. PMID- 11877470 TI - CCR3 is essential for skin eosinophilia and airway hyperresponsiveness in a murine model of allergic skin inflammation. AB - The CC chemokine receptor 3 (CCR3) is expressed by eosinophils, mast cells, and Th2 cells. We used CCR3(-/-) mice to assess the role of CCR3 in a murine model of allergic skin inflammation induced by repeated epicutaneous sensitization with ovalbumin (OVA), and characterized by eosinophil skin infiltration, local expression of Th2 cytokines, and airway hyperresponsiveness (AHR) to inhaled antigen. Eosinophils and the eosinophil product major basic protein were absent from the skin of sham and OVA-sensitized CCR3(-/-) mice. Mast cell numbers and expression of IL-4 mRNA were normal in skin of CCR3(-/-) mice, suggesting that CCR3 is not important for infiltration of the skin by mast cells and Th2 cells. CCR3(-/-) mice produced normal levels of OVA-specific IgE, and their splenocytes secreted normal amounts of IL-4 and IL-5 following in vitro stimulation with OVA, indicating effective generation of systemic Th2 helper responses. Recruitment of eosinophils to lung parenchyma and bronchoalveolar lavage (BAL) fluid was severely impaired in CCR3(-/-) mice, which failed to develop AHR to methacholine following antigen inhalation. These results suggest that CCR3 plays an essential role in eosinophil recruitment to the skin and the lung and in the development of AHR. PMID- 11877471 TI - Insulin signaling coordinately regulates cardiac size, metabolism, and contractile protein isoform expression. AB - To investigate the role of insulin signaling on postnatal cardiac development, physiology, and cardiac metabolism, we generated mice with a cardiomyocyte selective insulin receptor knockout (CIRKO) using cre/loxP recombination. Hearts of CIRKO mice were reduced in size by 20-30% due to reduced cardiomyocyte size and had persistent expression of the fetal beta-myosin heavy chain isoform. In CIRKO hearts, glucose transporter 1 (GLUT1) expression was reduced by about 50%, but there was a twofold increase in GLUT4 expression as well as increased rates of cardiac glucose uptake in vivo and increased glycolysis in isolated working hearts. Fatty acid oxidation rates were diminished as a result of reduced expression of enzymes that catalyze mitochondrial beta-oxidation. Although basal rates of glucose oxidation were reduced, insulin unexpectedly stimulated glucose oxidation and glycogenolysis in CIRKO hearts. Cardiac performance in vivo and in isolated hearts was mildly impaired. Thus, insulin signaling plays an important developmental role in regulating postnatal cardiac size, myosin isoform expression, and the switching of cardiac substrate utilization from glucose to fatty acids. Insulin may also modulate cardiac myocyte metabolism through paracrine mechanisms by activating insulin receptors in other cell types within the heart. PMID- 11877472 TI - Glatiramer acetate (Copaxone) therapy induces CD8(+) T cell responses in patients with multiple sclerosis. AB - Glatiramer acetate (GA; Copaxone) is a random copolymer of glutamic acid, lysine, alanine, and tyrosine that is used therapeutically in patients with multiple sclerosis (MS). To investigate the mechanism of the drug's immunomodulatory effect, we used immunophenotypic approaches to characterize the precise nature of GA-induced T cell responses. We demonstrate here that healthy individuals and untreated MS patients exhibit prominent T cell proliferative responses to GA. However, these responses are different in distinct subsets of T cells. Whereas GA induced CD4(+) T cell responses are comparable in healthy individuals and MS patients, CD8(+) T cell responses are significantly lower in untreated MS patients. Treatment with GA results in upregulation of these CD8(+) responses with restoration to levels observed in healthy individuals. Both CD4(+) and CD8(+) GA-specific responses are HLA-restricted. GA therapy also induces a change in the cytokine profile of GA-specific CD4(+) and CD8(+) T cells. This study provides the first direct immunophenotypic evidence, to our knowledge, of GA specific CD8(+) T cell responses and their upregulation during the course of therapy, which may suggest a role for these responses in the immunomodulatory effects of the drug. PMID- 11877473 TI - Provision of antigen and CD137 signaling breaks immunological ignorance, promoting regression of poorly immunogenic tumors. AB - Treatment of advanced, poorly immunogenic tumors in animal models, considered the closest simulation available thus far for conditions observed in cancer patients, remains a major challenge for cancer immunotherapy. We reported previously that established tumors in mice receiving an agonistic mAb to the T cell costimulatory molecule 4-1BB (CD137) regress due to enhanced tumor antigen-specific cytotoxic T lymphocyte responses. In this study, we demonstrate that several poorly immunogenic tumors, including C3 tumor, TC-1 lung carcinoma, and B16-F10 melanoma, once established as solid tumors or metastases, are refractory to treatment by anti-4-1BB mAb. We provide evidence that immunological ignorance, rather than anergy or deletion, of tumor antigen--specific CTLs during the progressive growth of tumors prevents costimulation by anti-4-1BB mAb. Breaking CTL ignorance by immunization with a tumor antigen-derived peptide, although insufficient to stimulate a curative CTL response, is necessary for anti--4-1BB mAb to induce a CTL response leading to the regression of established tumors. Our results suggest a new approach for immunotherapy of human cancers. PMID- 11877474 TI - Elastase-mediated phosphatidylserine receptor cleavage impairs apoptotic cell clearance in cystic fibrosis and bronchiectasis. AB - Cystic fibrosis is characterized by an early and sustained influx of inflammatory cells into the airways and by release of proteases. Resolution of inflammation is normally associated with the orderly removal of dying apoptotic inflammatory cells through cell recognition receptors, such as the phosphatidylserine receptor, CD36, and alpha v integrins. Accordingly, removal of apoptotic inflammatory cells may be impaired in persistent inflammatory responses such as that seen in cystic fibrosis airways. Examination of sputa from cystic fibrosis and non-cystic fibrosis bronchiectasis patients demonstrated an abundance of apoptotic cells, in excess of that seen in patients with chronic bronchitis. In vitro, cystic fibrosis and bronchiectasis airway fluid directly inhibited apoptotic cell removal by alveolar macrophages in a neutrophil elastase-dependent manner, suggesting that elastase may impair apoptotic cell clearance in vivo. Flow cytometry demonstrated that neutrophil elastase cleaved the phosphatidylserine receptor, but not CD36 or CD32 (Fc gamma RII). Cleavage of the phosphatidylserine receptor by neutrophil elastase specifically disrupted phagocytosis of apoptotic cells, implying a potential mechanism for delayed apoptotic cell clearance in vivo. Therefore, defective airway clearance of apoptotic cells in cystic fibrosis and bronchiectasis may be due to elastase mediated cleavage of phosphatidylserine receptor on phagocytes and may contribute to ongoing airway inflammation. PMID- 11877475 TI - Proteasome inhibition reduces superantigen-mediated T cell activation and the severity of psoriasis in a SCID-hu model. AB - There is increasing evidence that bacterial superantigens contribute to inflammation and T cell responses in psoriasis. Psoriatic inflammation entails a complex series of inductive and effector processes that require the regulated expression of various proinflammatory genes, many of which require NF-kappa B for maximal trans-activation. PS-519 is a potent and selective proteasome inhibitor based upon the naturally occurring compound lactacystin, which inhibits NF-kappa B activation by blocking the degradation of its inhibitory protein I kappa B. We report that proteasome inhibition by PS-519 reduces superantigen-mediated T cell activation in vitro and in vivo. Proliferation was inhibited along with the expression of very early (CD69), early (CD25), and late T cell (HLA-DR) activation molecules. Moreover, expression of E-selectin ligands relevant to dermal T cell homing was reduced, as was E-selectin binding in vitro. Finally, PS 519 proved to be therapeutically effective in a SCID-hu xenogeneic psoriasis transplantation model. We conclude that inhibition of the proteasome, e.g., by PS 519, is a promising means to treat T cell-mediated disorders such as psoriasis. PMID- 11877477 TI - Essential immunoregulatory role for BCAP in B cell development and function. AB - BCAP was recently cloned as a binding molecule to phosphoinositide 3-kinase (PI3K). To investigate the role of BCAP, mutant mice deficient in BCAP were generated. While BCAP-deficient mice are viable, they have decreased numbers of mature B cells and B1 B cell deficiency. The mice produce lower titers of serum immunoglobulin (Ig)M and IgG3, and mount attenuated responses to T cell- independent type II antigen. Upon B cell receptor cross-linking, BCAP-deficient B cells exhibit reduced Ca(2+) mobilization and poor proliferative responses. These findings demonstrate that BCAP plays a pivotal immunoregulatory role in B cell development and humoral immune responses. PMID- 11877476 TI - Treatment of primary HIV-1 infection with cyclosporin A coupled with highly active antiretroviral therapy. AB - Primary HIV-1 infection causes extensive immune activation, during which CD4(+) T cell activation supports massive HIV-1 production. We tested the safety and the immune-modulating effects of combining cyclosporin A (CsA) treatment with highly active antiretroviral therapy (HAART) during primary HIV-1 infection. Nine adults with primary HIV-1 infection were treated with CsA along with HAART. At week 8, all patients discontinued CsA but maintained HAART. Viral replication was suppressed to a comparable extent in the CsA + HAART cohort and in 29 control patients whose primary infection was treated with HAART alone. CsA restored normal CD4(+) T cell levels, both in terms of percentage and absolute numbers. The increase in CD4(+) T cells was apparent within a week and persisted throughout the study period. CsA was not detrimental to virus-specific CD8(+) or CD4(+) T cell responses. At week 48, the proportion of IFN-gamma-secreting CD4(+) and CD4(+)CCR7(-) T cells was significantly higher in the CsA + HAART cohort than in the HAART-alone cohort. In conclusion, rapid shutdown of T cell activation in the early phases of primary HIV-1 infection can have long-term beneficial effects and establish a more favorable immunologic set-point. Appropriate, immune-based therapeutic interventions may represent a valuable complement to HAART for treating HIV infection. PMID- 11877478 TI - Reciprocal roles for CCAAT/enhancer binding protein (C/EBP) and PU.1 transcription factors in Langerhans cell commitment. AB - Myeloid progenitor cells give rise to a variety of progenies including dendritic cells. However, the mechanism controlling the diversification of myeloid progenitors into each progeny is largely unknown. PU.1 and CCAAT/enhancing binding protein (C/EBP) family transcription factors have been characterized as key regulators for the development and function of the myeloid system. However, the roles of C/EBP transcription factors have not been fully identified because of functional redundancy among family members. Using high titer--retroviral infection, we demonstrate that a dominant-negative C/EBP completely blocked the granulocyte--macrophage commitment of human myeloid progenitors. Alternatively, Langerhans cell (LC) commitment was markedly facilitated in the absence of tumor necrosis factor (TNF)alpha, a strong inducer of LC development, whereas expression of wild-type C/EBP in myeloid progenitors promoted granulocytic differentiation, and completely inhibited TNFalpha-dependent LC development. On the other hand, expression of wild-type PU.1 in myeloid progenitors triggered LC development in the absence of TNFalpha, and its instructive effect was canceled by coexpressed C/EBP. Our findings establish reciprocal roles for C/EBP and PU.1 in LC development, and provide new insight into the molecular mechanism of LC development, which has not yet been well characterized. PMID- 11877479 TI - Toll-like receptor 4 resides in the Golgi apparatus and colocalizes with internalized lipopolysaccharide in intestinal epithelial cells. AB - Toll-like receptor (TLR) 4 is mainly found on cells of the myelopoietic lineage. It recognizes lipopolysaccharide (LPS) and mediates cellular activation and production of proinflammatory cytokines. Less is known about the distribution and role of TLR4 in epithelial cells that are continuously exposed to microbes and microbial products. Here we show that the murine small intestinal epithelial cell line m-IC(cl2) is highly responsive to LPS and expresses both CD14 and TLR4. Transcription and surface membrane staining for CD14 were up-regulated upon LPS exposure. Surprisingly, TLR4 immunostaining revealed a strictly cytoplasmic paranuclear distribution. This paranuclear compartment could be identified as the Golgi apparatus. LPS added to the supernatant was internalized by m-IC(cl2) cells and colocalized with TLR4. Continuous exposure to LPS led to a tolerant phenotype but did not alter TLR4 expression nor cellular distribution. Thus, intestinal epithelial cells might be able to provide the initial proinflammatory signal to attract professional immune cells to the side of infection. The cytoplasmic location of TLR4, which is identical to the final location of internalized LPS, further indicates an important role of cellular internalization and cytoplasmic traffic in the process of innate immune recognition. PMID- 11877480 TI - Structure of a complex of the human alpha/beta T cell receptor (TCR) HA1.7, influenza hemagglutinin peptide, and major histocompatibility complex class II molecule, HLA-DR4 (DRA*0101 and DRB1*0401): insight into TCR cross-restriction and alloreactivity. AB - The alpha/beta T cell receptor (TCR) HA1.7 specific for the hemagglutinin (HA) antigen peptide from influenza A virus is HLA-DR1 restricted but cross-reactive for the HA peptide presented by the allo-major histocompatibility complex (MHC) class II molecule HLA-DR4. We report here the structure of the HA1.7/DR4/HA complex, determined by X-ray crystallography at a resolution of 2.4 A. The overall structure of this complex is very similar to the previously reported structure of the HA1.7/DR1/HA complex. Amino acid sequence differences between DR1 and DR4, which are located deep in the peptide binding groove and out of reach for direct contact by the TCR, are able to indirectly influence the antigenicity of the pMHC surface by changing the conformation of HA peptide residues at position P5 and P6. Although TCR HA1.7 is cross-reactive for HA presented by DR1 and DR4 and tolerates these conformational differences, other HA specific TCRs are sensitive to these changes. We also find a dependence of the width of the MHC class II peptide-binding groove on the sequence of the bound peptide by comparing the HA1.7/DR4/HA complex with the structure of DR4 presenting a collagen peptide. This structural study of TCR cross-reactivity emphasizes how MHC sequence differences can affect TCR binding indirectly by moving peptide atoms. PMID- 11877481 TI - Cysteinyl leukotrienes and uridine diphosphate induce cytokine generation by human mast cells through an interleukin 4-regulated pathway that is inhibited by leukotriene receptor antagonists. AB - We previously reported that interleukin (IL)-4 upregulates the expression of leukotriene C(4) synthase (LTC(4)S) by human cord blood--derived mast cells (hMCs), augments their high-affinity Fc receptor for IgE (Fc(epsilon)RI) dependent generation of eicosanoids and cytokines, and induces a calcium flux in response to cysteinyl leukotrienes (cys-LTs) and uridine diphosphate (UDP) that is blocked by cys-LT receptor antagonists. We speculated that this IL-4 dependent, receptor-mediated response to the cys-LTs and UDP might induce cytokine generation by hMCs without concomitant exocytosis. Unlike hMCs maintained in cytoprotective stem cell factor (SCF) alone, hMCs primed for 5 d with IL-4 responded to UDP (1microM), LTC(4) (100 nM), and LTD(4) (100 nM) by producing IL-5, tumor necrosis factor (TNF)-alpha, and especially large quantities of macrophage inflammatory protein (MIP)-1beta de novo at 6 h, preceded by the induced expression of the corresponding mRNAs. Cys-LT- and UDP mediated cytokine production by the primed hMCs occurred without histamine release or PGD(2) generation and was inhibited by the CysLT1 receptor antagonist MK571. Additionally, pretreatment of hMCs with MK571 or with the cys-LT biosynthetic inhibitor MK886 decreased IL-5 and TNF-alpha production in response to IgE receptor cross-linkage, implying a positive feedback by endogenously produced cys-LTs. Cys-LTs and UDP thus orchestrate a novel, IL-4-regulated, non IgE-dependent hMC activation for cytokine gene induction that could be initiated by microbes, cellular injury, or neurogenic or inflammatory signals; and this pathobiologic event would not be recognized in tissue studies where hMC activation is classically defined by exocytosis. PMID- 11877482 TI - Lipid raft microdomains: a gateway for compartmentalized trafficking of Ebola and Marburg viruses. AB - Spatiotemporal aspects of filovirus entry and release are poorly understood. Lipid rafts act as functional platforms for multiple cellular signaling and trafficking processes. Here, we report the compartmentalization of Ebola and Marburg viral proteins within lipid rafts during viral assembly and budding. Filoviruses released from infected cells incorporated raft-associated molecules, suggesting that viral exit occurs at the rafts. Ectopic expression of Ebola matrix protein and glycoprotein supported raft-dependent release of filamentous, virus-like particles (VLPs), strikingly similar to live virus as revealed by electron microscopy. Our findings also revealed that the entry of filoviruses requires functional rafts, identifying rafts as the site of virus attack. The identification of rafts as the gateway for the entry and exit of filoviruses and raft-dependent generation of VLPs have important implications for development of therapeutics and vaccination strategies against infections with Ebola and Marburg viruses. PMID- 11877483 TI - In vitro generation of interleukin 10-producing regulatory CD4(+) T cells is induced by immunosuppressive drugs and inhibited by T helper type 1 (Th1)- and Th2-inducing cytokines. AB - We show that a combination of the immunosuppressive drugs, vitamin D3 and Dexamethasone, induced human and mouse naive CD4(+) T cells to differentiate in vitro into regulatory T cells. In contrast to the previously described in vitro derived CD4(+) T cells, these cells produced only interleukin (IL)-10, but no IL 5 and interferon (IFN)-gamma, and furthermore retained strong proliferative capacity. The development of these IL-10-producing cells was enhanced by neutralization of the T helper type 1 (Th1)- and Th2-inducing cytokines IL-4, IL 12, and IFN-gamma. These immunosuppressive drugs also induced the development of IL-10-producing T cells in the absence of antigen-presenting cells, with IL-10 acting as a positive autocrine factor for these T cells. Furthermore, nuclear factor (NF)-kappaB and activator protein (AP)-1 activities were inhibited in the IL-10-producing cells described here as well as key transcription factors involved in Th1 and Th2 subset differentiation. The regulatory function of these in vitro generated IL-10-producing T cells was demonstrated by their ability to prevent central nervous system inflammation, when targeted to the site of inflammation, and this function was shown to be IL-10 dependent. Generating homogeneous populations of IL-10-producing T cells in vitro will thus facilitate the use of regulatory T cells in immunotherapy. PMID- 11877484 TI - Natural killer T cell ligand alpha-galactosylceramide enhances protective immunity induced by malaria vaccines. AB - The important role played by CD8(+) T lymphocytes in the control of parasitic and viral infections, as well as tumor development, has raised the need for the development of adjuvants capable of enhancing cell-mediated immunity. It is well established that protective immunity against liver stages of malaria parasites is primarily mediated by CD8(+) T cells in mice. Activation of natural killer T (NKT) cells by the glycolipid ligand, alpha-galactosylceramide (alpha-GalCer), causes bystander activation of NK, B, CD4(+), and CD8(+) T cells. Our study shows that coadministration of alpha-GalCer with suboptimal doses of irradiated sporozoites or recombinant viruses expressing a malaria antigen greatly enhances the level of protective anti-malaria immunity in mice. We also show that coadministration of alpha-GalCer with various different immunogens strongly enhances antigen-specific CD8(+) T cell responses, and to a lesser degree, Th1 type responses. The adjuvant effects of alpha-GalCer require CD1d molecules, Valpha14 NKT cells, and interferon gamma. As alpha-GalCer stimulates both human and murine NKT cells, these findings should contribute to the design of more effective vaccines against malaria and other intracellular pathogens, as well as tumors. PMID- 11877485 TI - Functionally distinct subsets of CD1d-restricted natural killer T cells revealed by CD1d tetramer staining. AB - CD1d-restricted natural killer (NK)T cells are known to potently secrete T helper (Th)1 and Th2 cytokines and to mediate cytolysis, but it is unclear how these contrasting functional activities are regulated. Using lipid antigen-loaded CD1d tetramers, we have distinguished two subsets of CD1d-restricted T cells in fresh peripheral blood that differ in cytokine production and cytotoxic activation. One subset, which was CD4(-), selectively produced the Th1 cytokines interferon gamma and tumor necrosis factor alpha, and expressed NKG2d, a marker associated with cytolysis of microbially infected and neoplastic cells. This subset up-regulated perforin after exposure to interleukin (IL)-2 or IL-12. In contrast, CD4(+) CD1d restricted NKT cells potently produced both Th1 and Th2 cytokines, up-regulated perforin in response to stimulation by phorbol myristate acetate and ionomycin but not IL-2 or IL-12, and could be induced to express CD95L. Further, for both CD1d-restricted NKT cell subsets, we found that antigenic stimulation induced cytokine production but not perforin expression, whereas exposure to inflammatory factors enhanced perforin expression but did not stimulate cytokine production. These results show that the various activities of CD1d-restricted T cells in tumor rejection, autoimmune disease, and microbial infections could result from activation of functionally distinct subsets, and that inflammatory and antigenic stimuli may influence different effector functions. PMID- 11877486 TI - Distinct functional lineages of human V(alpha)24 natural killer T cells. AB - CD1d-restricted autoreactive natural killer (NK)T cells have been reported to regulate a range of disease conditions, including type I diabetes and immune rejection of cancer, through the secretion of either T helper (Th)2 or Th1 cytokines. However, mechanisms underlying Th2 versus Th1 cytokine secretion by these cells are not well understood. Since most healthy subjects express <1 NKT cell per 1,000 peripheral blood lymphocytes (PBLs), we devised a new method based on the combined used of T cell receptor (TCR)-specific reagents alpha galactosylceramide (alphaGalCer) loaded CD1d-tetramers and anti-V(alpha)24 monoclonal antibody, to specifically identify and characterize these rare cells in fresh PBLs. We report here that CD4(+) and CD4(-)CD8(-) (double negative [DN]) NKT cell subsets represent functionally distinct lineages with marked differences in their profile of cytokine secretion and pattern of expression of chemokine receptors, integrins, and NK receptors. CD4(+) NKT cells were the exclusive producers of interleukin (IL)-4 and IL-13 upon primary stimulation, whereas DN NKT cells had a strict Th1 profile and prominently expressed several NK lineage receptors. These findings may explain how NKT cells could promote Th2 responses in some conditions and Th1 in others, and should be taken into consideration for intervention in relevant diseases. PMID- 11877487 TI - Specificity of tissue transglutaminase explains cereal toxicity in celiac disease. AB - Celiac disease is caused by a selective lack of T cell tolerance for gluten. It is known that the enzyme tissue transglutaminase (tTG) is involved in the generation of T cell stimulatory gluten peptides through deamidation of glutamine, the most abundant amino acid in gluten. Only particular glutamine residues, however, are modified by tTG. Here we provide evidence that the spacing between glutamine and proline, the second most abundant amino acid in gluten, plays an essential role in the specificity of deamidation. On the basis of this, algorithms were designed and used to successfully predict novel T cell stimulatory peptides in gluten. Strikingly, these algorithms identified many similar peptides in the gluten-like hordeins from barley and secalins from rye but not in the avenins from oats. The avenins contain significantly lower percentages of proline residues, which offers a likely explanation for the lack of toxicity of oats. Thus, the unique amino acid composition of gluten and related proteins in barley and rye favors the generation of toxic T cell stimulatory gluten peptides by tTG. This provides a rationale for the observation that celiac disease patients are intolerant to these cereal proteins but not to other common food proteins. PMID- 11877488 TI - Rapid cytotoxic T lymphocyte activation occurs in the draining lymph nodes after cutaneous herpes simplex virus infection as a result of early antigen presentation and not the presence of virus. AB - Localized cutaneous herpes simplex virus type 1 (HSV-1) infection leads to arming and initial expansion of cytotoxic T lymphocytes (CTLs) in the draining popliteal lymph nodes (PLNs) followed by migration and further proliferation in the spleen. To accurately characterize the sequence of events involved in the activation and generation of anti-HSV CTLs, we used T cell receptor (TCR) transgenic mice specific for the immunodominant epitope from HSV glycoprotein B (gB(498-505)). We describe the detection of the initiation of antigen presentation in the draining lymph nodes by 4-6 h after infection with HSV-1. Analysis of CD69 up-regulation revealed activation of gB-specific CD8(+) T cells by 6-8 h after infection. Furthermore, we show that T cell proliferation begins no sooner than 24 h after activation and is marked by the concurrent appearance of CTL activity in the PLNs. These events are not dependent on the presence of virus in the draining lymph nodes, and suggest a requirement for recruitment of professional antigen presenting cells to the site of T cell activation. Consequently, we have defined the initiation of the CD8(+) T cell-mediated response to cutaneous HSV-1 infection, demonstrating that the immune response to localized viral infection depends only on the appearance of cells presenting virus-derived antigen and commences with remarkable swiftness. PMID- 11877489 TI - Estimating the precursor frequency of naive antigen-specific CD8 T cells. AB - The constraint of fitting a diverse repertoire of antigen specificities in a limited total population of lymphocytes results in the frequency of naive cells specific for any given antigen (defined as the precursor frequency) being below the limit of detection by direct measurement. We have estimated this precursor frequency by titrating a known quantity of antigen-specific cells into naive recipients. Adoptive transfer of naive antigen-specific T cell receptor transgenic cells into syngeneic nontransgenic recipients, followed by stimulation with specific antigen, results in activation and expansion of both donor and endogenous antigen-specific cells in a dose-dependent manner. The precursor frequency is equal to the number of transferred cells when the transgenic and endogenous responses are of equal magnitude. Using this method we have estimated the precursor frequency of naive CD8 T cells specific for the H-2D(b)-restricted GP33-41 epitope of LCMV to be 1 in 2 x 10(5). Thus, in an uninfected mouse containing approximately 2-4 x 10(7) naive CD8 T cells we estimate there to be 100-200 epitope-specific cells. After LCMV infection these 100-200 GP33-specific naive CD8 T cells divide >14 times in 1 wk to reach a total of approximately 10(7) cells. Approximately 5% of these activated GP33-specific effector CD8 T cells survive to generate a memory pool consisting of approximately 5 x 10(5) cells. Thus, an acute LCMV infection results in a >1,000-fold increase in precursor frequency of D(b)GP33-specific CD8 T cells from 2 x 10(2) naive cells in uninfected mice to 5 x 10(5) memory cells in immunized mice. PMID- 11877491 TI - Synaptic mechanisms regulating the activation of a Ca(2+)-mediated plateau potential in developing relay cells of the LGN. AB - Using intracellular recordings in an isolated (in vitro) rat brain stem preparation, we examined the synaptic responses of developing relay neurons in the dorsal lateral geniculate nucleus (LGN). In newborn rats, strong stimulation of the optic tract (OT) evoked excitatory postsynaptic potentials (EPSPs) that gave rise to a sustained (300-1,300 ms), slow-decaying (<0.01 mV/s), depolarization (25-40 mV). Riding atop this response was a train of spikes of variable amplitude. We refer to this synaptically evoked event as a plateau potential. Pharmacology experiments indicate the plateau potential was mediated by the activation of high-threshold L-type Ca(2+) channels. Synaptic activation of the plateau potential relied on N-methyl-D-aspartate (NMDA) receptor-mediated activity and the spatial and/or temporal summation of retinally evoked EPSPs. Inhibitory postsynaptic responses (IPSPs) did not prevent the expression of the plateau potential. However, GABA(A) receptor activity modulated the intensity of optic tract stimulation needed to evoke the plateau potential, while GABA(B) receptor activity affected its duration. Expression of the plateau potential was developmentally regulated, showing a much higher incidence at P1-2 (90%) than at P19-20 (1%). This was in part due to the fact that developing relay cells show a greater degree of spatial summation than their mature counterparts, receiving input from as many as 7-12 retinal ganglion cells. Early spontaneous retinal activity is also likely to trigger the plateau potential. Repetitive stimulation of optic tract in a manner that approximated the high-frequency discharge of retinal ganglion cells led to a massive temporal summation of EPSPs and the activation of a sustained depolarization (>1 min) that was blocked by L-type Ca(2+) channel antagonists. These age-related changes in Ca(2+) signaling may contribute to the activity-dependent refinement of retinogeniculate connections. PMID- 11877490 TI - Adjuvants of immunity: harnessing innate immunity to promote adaptive immunity. PMID- 11877492 TI - Position sensitivity of human muscle spindles: single afferent and population representations. AB - The representation of joint position at rest and during movement was investigated in 44 muscle spindle primary afferents originating from the extensor carpi radialis brevis (ECRb) and extensor digitorum (ED) of normal human subjects. Position sensitivity was estimated for each afferent, and 43 of 44 were position sensitive. In each trial, six sequential ramp-and-hold movements (2-6 degrees, 2 degrees/s, total 24 degrees) flexed the relaxed wrist, beginning from the angle at which the afferent was just recruited. Joint position was represented by three specific features of afferent firing patterns: the steady-state firing rate during the 4-s hold period between ramps, the initial burst at the beginning of each ramp, and the ramp increase in firing rate later in the movement. The position sensitivity of the initial burst (1.27 +/- 0.90 pps/degree, mean +/- SD) was several times higher than that of the hold period (0.40 +/- 0.30 pps/degree) and not different from that of the ramp increase in firing rate (1.36 +/- 0.68 pps/degree). The wrist position sensitivities of ECRb and ED afferents were equivalent, as were their recruitment angles and angular ranges of position sensitivity. Muscle spindle afferents, both individually and as a population, were shown to represent static joint position via the hold rate and the initial burst. Afferents were recruited over the entire 110 degree range of wrist positions investigated; however, the angular range over which each feature represented joint position was extremely limited (approximately 15 degrees). The population response, based on the summed activity of the 43 afferents, was monotonically related to joint position, and it was strongly influenced by the afferent recruitment pattern, but less so by the position sensitivities of the individual afferents. PMID- 11877493 TI - GABA(B) presynaptic inhibition has an in vivo time constant sufficiently rapid to allow modulation at theta frequency. AB - Cyclical activity of GABAergic interneurons during theta rhythm could mediate phasic changes in strength of glutamatergic synaptic transmission in the hippocampal formation if presynaptic inhibition from activation of GABA(B) receptors is sufficiently rapid to change within a theta cycle. The experiments described here analyzed the time course of GABA(B) modulation using a heterosynaptic depression paradigm in anesthetized rats at physiological temperatures. Heterosynaptic depression of the slope of evoked potentials decayed with a time constant that would allow significant changes in transmission across different phases of the theta cycle. This heterosynaptic depression was significantly reduced by local infusion of the GABA(B) receptor antagonist CGP55845A. PMID- 11877494 TI - Effects of methylphenidate on the membrane potential and current in neurons of the rat locus coeruleus. AB - Effects of methylphenidate (MPH), a therapeutic agent used in children presenting the attention deficit hyperactivity disorder (ADHD), on the membrane potential and current in neurons of the rat locus coeruleus (LC) were examined using intracellular and whole cell patch-clamp recording techniques. Application of MPH (30 microM) to artificial cerebrospinal fluid (ACSF) produced a hyperpolarizing response with amplitude of 12 +/- 1 mV (n = 29). Spontaneous firing of LC neurons was blocked during the MPH-induced hyperpolarization. Superfusion of LC neurons with ACSF containing 0 mM Ca(2+) and 11 mM Mg(2+) (Ca(2+)-free ACSF) produced a depolarizing response associated with an increase in spontaneous firing of the action potential. The MPH-induced hyperpolarization was blocked in Ca(2+)-free ACSF. Yohimbine (1 microM) and prazosin (10 microM), antagonists for alpha(2) and alpha(2B/2C) receptors, respectively, blocked the MPH-induced hyperpolarization in LC neurons. Tetrodotoxin (TTX, 1 microM) produced a partial depression of the MPH-induced hyperpolarization in LC neurons. Under the whole cell patch-clamp condition, MPH (30-300 microM) produced an outward current (I(MPH)) with amplitude of 110 +/- 6 pA (n = 17) in LC neurons. The I(MPH) was blocked by Co(2+) (1 mM). During prolonged application of MPH (300 microM for 45 min), the hyperpolarization gradually decreased in the amplitude and eventually disappeared, possibly because of depression of norepinephrine (NE) release from noradrenergic nerve terminals. At a low concentration (1 microM), MPH produced no outward current but consistently enhanced the outward current induced by NE. These results suggest that the MPH-induced response is mediated by NE via alpha(2B/2C)-adrenoceptors in LC neurons. I(MPH) was associated with an increase in the membrane conductance of LC neurons. The I(MPH) reversed its polarity at 102 +/- 6 mV (n = 8) in the ACSF. The reversal potential of I(MPH) was changed by 54 mV per decade change in the external K(+) concentration. Current-voltage relationship showed that the I(MPH) exhibited inward rectification. Ba(2+) (100 microM) suppressed the amplitude and the inward rectification of the I(MPH.) These results suggest that the I(MPH) is produced by activation of inward rectifier K(+) channels in LC neurons. PMID- 11877495 TI - Uniform range of conduction times from the lateral amygdala to distributed perirhinal sites. AB - Much data indicate that the perirhinal (PRH) cortex plays a critical role in declarative memory and that the amygdala facilitates this process under emotionally arousing conditions. However, assuming that the amygdala does so by promoting Hebbian interactions in the PRH cortex is hard to reconcile with the fact that variable distances separate amygdala neurons from their PRH projection sites. Indeed, to achieve a synchronized activation of distributed PRH sites, amygdala axons should display a uniform range of conduction times, irrespective of distance to target. To determine if amygdala axons meet this condition, we measured the antidromic response latencies of lateral amygdala (LA) neurons to electrical stimuli delivered at various rostrocaudal levels of the PRH cortex in cats anesthetized with isoflurane. Although large variations in antidromic response latencies were observed, they were unrelated to the distance between the PRH stimulation sites and LA neurons. To determine whether this result was an artifact due to current spread, two control experiments were performed. First, we examined the antidromic response latency of intrinsic PRH neurons. Although we used the same methods as in the first experiment, the antidromic response latency of PRH neurons to electrical stimuli applied in the PRH cortex increased linearly with the distance between the stimulating and recording sites. Second, we measured the antidromic response latency of PRH neurons projecting to the LA. In this pathway, we also found a statistically significant correlation between conduction times and distance to target. Thus these results support the intriguing possibility that the conduction velocity and/or trajectory of LA axons are adjusted to compensate for variations in distance between the LA and distinct rostrocaudal PRH sites. We hypothesize that because of their uniform range of conduction times to the PRH cortex, LA neurons can generate short time windows of depolarization facilitating Hebbian associations between coincident, but spatially distributed, activity patterns in the PRH cortex. In this context, the temporal scatter of conduction times in the LA to PRH pathway is conceived as a mechanism used to lengthen the period of depolarization to compensate for conduction delays within intrinsic PRH pathways. In part, this mechanism might explain how the amygdala promotes memory storage in emotionally arousing conditions. PMID- 11877497 TI - Direct and indirect actions of dopamine on the membrane potential in medium spiny neurons of the mouse neostriatum. AB - Many studies have shown dopamine (DA) to have a modulatory effect on neuronal excitability, which cannot be simply classified as excitatory or inhibitory in the neostriatum. To clarify whether the responses to DA (10-30 microM) are excitatory or inhibitory in the mouse medium spiny neurons, we examined the effects of DA agonists on the synchronous potential trajectory from the resting potential to the subthreshold potential. The DA-induced potential changes, which were estimated at the subthreshold potential (approximately -60 mV), were summarized as the combination of three kinds of responses: an initial hyperpolarization lasting approximately 1 min and a slow depolarization and/or hyperpolarization lasting more than 20 min. A D(1)-like receptor agonist, R(+)-6 chloro-7,8-dihydroxy-1-phenyl-2,3,4,5-tetrahydro-1H-3-benzazepine hydrobromide (SKF81297, 1 microM) mainly induced the initial hyperpolarization and slow depolarization. A D(2)-like receptor agonist, trans-(-)-4aR-4,4a,5,6,7,8,8a,9 octahydro-5-propyl-1H-pyrazolo[3,4-g]quinoline hydrochloride (quinpirole, 1 microM), mainly induced the initial hyperpolarization and slow hyperpolarization. D(1)-like receptor antagonist R(+)-7-chloro-8-hydroxy-3-methyl1-phenyl-2,3,4,5 tetrahydro-1H-3-benzazepine hydrochloride (SCH23390, 1 microM) depressed both the initial hyperpolarization and slow depolarization. D(2)-like receptor antagonist sulpiride (1 microM) depressed all the DA-induced responses except for the slow depolarization. TTX (0.5 microM) abolished all the DA-induced responses. Bicuculline (20 microM) and atropine (1 microM) abolished the DA-induced initial hyperpolarization and slow depolarization, respectively. Either DL-2-amino-5 phosphonopentanoic acid (AP5; 100 microM) or 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX, 20 microM) blocked both the initial hyperpolarization and slow depolarization. The application of exogenous glutamate (Glu) mimicked the initial hyperpolarization and slow depolarization. These results suggest that the initial hyperpolarization is mainly due to GABA release via the cooperative action of D(1)- and D(2)-like receptors and Glu receptors in GABAergic interneurons, whereas the slow depolarization is mediated by acetylcholine (ACh) release via the cooperative action of mainly D(1)-like receptors and Glu receptors in cholinergic interneurons. The potential oscillation was generated at the subthreshold level in a Ba(2+)-, AP5-, CNQX-, bicuculline-, and atropine containing medium. The oscillation depressed after the addition of TTX, Co(2+), or DA. In DA agonists, quinpirole rather than SKF81297 had a more depressive effect on the potential oscillation. These results indicate that the slow hyperpolarization is due to the suppression of noninactivating Na(+)-Ca(2+) conductances via mainly D(2)-like receptors in the medium spiny neurons. In conclusion, the DA actions on the medium spiny neurons show a transient inhibition by the activation of D(1)- and D(2)-like receptors in mainly GABAergic interneurons and a tonic excitation and/or inhibition by the activation of mainly D(1)-like receptors in cholinergic interneurons and by the activation of mainly D(2)-like receptors in the medium spiny neurons, respectively. PMID- 11877496 TI - Synchronous and asynchronous exocytosis induced by subthreshold high K+ at Cs(+) loaded terminals of rat hippocampal neurons. AB - Transmitter release at Cs(+)-loaded autaptic terminals was selectively activated by the subthreshold concentration of external K+, and Ca(2+) channel types and transmitter pools involved in synchronous and asynchronous exocytosis were studied. When a neuron was depolarized to +30 mV by applying a current through a pipette containing Cs(+) for >30 s, a rapid external K+ jump to 3.75-10 mM, otherwise ineffective, produced an outward current (K10 response). K10 responses were initially graded (type-1) and then became a spike and plateau-shape with (type-2) or without a latency (type-3). On repolarization to -60 mV, a high K+ jump induced inward currents (called also K10 response) similar to those at +30 mV, whose shape changed from that of type-3, then type-2 and finally type-1 over 30 min. During a period favorable for inducing a type-3 response, a current similar to this response was generated by a voltage pulse (+ 80 or 90 mV, 20 or 30 ms) to the cell soma. Currents similar to K10 responses were rarely induced by a high K+ jump without a conditioning depolarization except for some cells, but consistently produced when 3 mM Cs(+) and 50 microM 4-aminopyridine were externally applied for tens of minutes. Picrotoxin, 6-cyano-7-nitroquinoxaline 2,3-dione with 3-[(RS)-2-carboxypiperazin-4-yl]-propyl-1-phosphonic acid or Cd(2+) in, or Ca(2+) removal from, a high-K+ solution blocked all the K10 responses, while a plateau remaining after a high K+ jump was not blocked by Ca(2+) removal immediately after the K+ jump. Thus Cs(+) loading and decreased K+ concentration in autaptic terminals by a conditioning depolarizing current selectively sensitize the terminals to a subthreshold high K+ jump for depolarization to activate synchronous or asynchronous transmitter release. Nicardipine (5-10 microM) blocked type-1 and -2 responses but not type-3 responses, while omega-conotoxin (10 microM) blocked all the types of K10 response in the presence of nicardipine. Increasing the interval of high K+ jumps biphasically increased the magnitude of K10 response, preferentially in the postjump fraction reflecting purely the asynchronous activation of exocytotic machinery, and decreased the reduction of miniature postsynaptic current frequency after a K10 response. These results suggest the roles of N(P/Q)-type Ca(2+) channels in synchronous exocytosis at the terminals, L-type Ca(2+) channels in initiating a Ca(2+) action potential at the parent axon and both types in asynchronous exocytosis and also suggest the different releasable pools of transmitter for two modes of exocytosis in cultured hippocampal neurons. PMID- 11877498 TI - Activation of embryonic red and white muscle fibers during fictive swimming in the developing zebrafish. AB - Sub-threshold, motoneuron-evoked synaptic activity was observed in zebrafish embryonic red (ER) and white (EW) muscle fibers paralyzed with a dose of D tubocurarine insufficient to abolish synaptic activity to determine whether muscle activation was coordinated to produce the undulating body movements required for locomotion. Paired whole-cell recordings revealed a synaptic drive that alternated between ipsilateral and contralateral myotomes and exhibited a rostral-caudal delay in timing appropriate for swimming. Both ER and EW muscle were activated during fictive swimming. However, at the fastest fictive swimming rates, ER fibers were de-recruited, whereas they could be active in isolation of EW fibers at the slowest fictive swimming rates. Prior to hatching, fictive swimming was preceded by a lower frequency, more robust and rhythmic synaptic drive resembling the "coiling" behavior of fish embryos. The motor activity observed in paralyzed zebrafish closely resembled the swimming and coiling behaviors observed in these developing fishes. At the early developmental stages examined in this study, myotomal muscle recruitment and coordination were similar to that observed in adult fishes during swimming. Our results indicate that the patterned activation of myotomal muscle is set from the onset of development. PMID- 11877499 TI - Selective suppression of late laryngeal adductor responses by N-methyl-D aspartate receptor blockade in the cat. AB - Laryngeal adductor responses to afferent stimulation play a key role in airway protection. Although vital for protection during cough and swallow, these responses also must be centrally controlled to prevent airway obstruction by laryngospasm during prolonged stimulation. Our purpose was to determine the role of N-methyl-D-aspartate (NMDA) receptors in modulating early R1 responses (at 9 ms) and/or later more prolonged R2 responses (at 36 ms) during electrical stimulation of the laryngeal afferent fibers contained in the internal branch of the superior laryngeal nerve in the cat. The percent occurrence, amplitude, and conditioning of muscle responses to single superior laryngeal nerve (SLN) stimuli presented in pairs at interstimulus intervals of 250 ms were measured in three experiments: 1) animals that had ketamine as anesthetic premedication were compared with those who did not, when both were maintained under alpha-chloralose anesthesia. 2) The effects of administering ketamine in one group of animals were compared with increasing the depth of alpha-chloralose anesthesia without NMDA receptor blockade in another group of animals. 3) The effects of dextromethorphan (without anesthetic effects) were examined in another group of animals. In the first experiment, the occurrence of R2 responses were reduced from 95% in animals without ketamine premedication to 25% in animals with ketamine premedication (P = 0.015). No differences occurred in the occurrence, amplitude, latency, or conditioning effects on R1 responses between these groups. In the second experiment, the occurrence of R2 responses was reduced from 96 to 79% after an increase in the depth of anesthesia with alpha-chloralose in contrast with reductions in R2 occurrence from 98 to 19% following the administration of ketamine to induce NMDA receptor blockade along with increased anesthesia (P = 0.025). In the third experiment, R2 occurrence was reduced from 89 to 27% (P = 0.017) with administration of dextromethorphan while R1 response occurrence and amplitude did not change. In each of these experiments, NMDA receptor blockade did not have significant effects on cardiac or respiratory rates in any of the animals. The results demonstrate that NMDA receptors play an essential role in long latency R2 laryngeal responses to laryngeal afferent stimulation. On the other hand, early R1 laryngeal adductor responses are likely to involve non-NMDA receptor activation. PMID- 11877500 TI - Silent synapses in developing cerebellar granule neurons. AB - Silent synapses are excitatory synapses endowed exclusively with N-methyl-D aspartate (NMDA) responses that have been proposed to acquire alpha-amino-3 hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) responses during development and after long-term potentiation (LTP). These synapses are functionally silent because of the Mg(2+) block of NMDA receptors at resting potentials. Here we provide evidence for the presence of silent synapses in developing cerebellar granule cells. Using the patch-clamp technique in the whole-cell configuration, we recorded the spontaneous excitatory postsynaptic currents (sEPSCs) from rat cerebellar granule cells in culture and in slices at physiological concentration of Mg(2+) (1 mM). A holding potential of +60 mV removes Mg(2+) block of NMDA channels, allowing us to record NMDA-sEPSCs. We thus compared the frequency of AMPA-sEPSCs, recorded at -60 mV, with that of NMDA-sEPSCs, recorded at +60 mV. NMDA-sEPSCs occurred at higher frequency than the AMPA-sEPSCs in most cells recorded in slices from rats at postnatal day (P) <13 and in culture at 6-8 days after plating (DIV6-8). In a few cells from young rats (P6-9) and in most neurons in culture at DIV6 we recorded exclusively NMDA-sEPSCs, supporting the hypothesis of existence of functional synapses with NMDA and without AMPA receptors. Increasing glutamate release in the slice with cyclothiazide and temperature increased AMPA and NMDA-sEPSCs frequencies but failed to alter the relative ratio of frequency of occurrence. Frequency ratio of NMDA versus AMPA-sEPSCs in slices was correlated with the weighted time constant of decay (tau(w)) of NMDA-sEPSCs and decreased with development along the reported decrease of tau(w). We suggest that the prevalence of synaptic NR2A subunits that confer faster kinetics is paralleled by the disappearance of silent synapses early in cerebellar development. PMID- 11877501 TI - Sodium channel blocking actions of the kappa-opioid receptor agonist U50,488 contribute to its visceral antinociceptive effects. AB - The goal of the present study was to determine whether the kappa-opioid receptor agonist (ORA) U50,488 attenuates behavioral and primary afferent nerve responses to noxious colorectal distension (CRD) by sodium channel blockade. We tested the analgesic kappa-ORA (+/-)-trans U50,488, its enantiomers (-)-trans (1S,2S) U50,488 and non kappa-ORA (+)-trans (1R,2R)-U50,488, and/or its diastereomer (-) cis (1S,2R)-U50,488 for their ability to attenuate visceromotor and pelvic nerve afferent fiber responses to noxious CRD in vivo and voltage-activated sodium current in colon sensory neurons in vitro. In unanesthetized rats, subcutaneous administration of U50,488, (1S,2S)-U50,488, and (1R,2R)-U50,488 attenuated the behavioral visceromotor response to noxious CRD; the rank order of potency was: (1S,2S)-U50,488 > U50,488 angle quotation mark, right (1R,2R)-U50,488. U50,488 and its stereoisomers also inhibited responses of decentralized pelvic nerve afferent fibers to noxious CRD in a dose-dependent manner. Cumulative doses of 16 mg/kg of (1S,2S)-U50,488, (1S,2R)-U50,488, and (1R,2R)-U50,488 reduced responses to a mean 29, 30, and 47% of control, respectively. The mean inhibitory doses of these drugs were not different (range: 6.6-10.8 mg/kg). Sodium channel blockers mexiletine and carbamazepine mimicked the effect of U50,488. In contrast, the kappa-ORAs dynorphin (1-13) and ICI 204,488 were ineffective in attenuating pelvic nerve activity. Perfusion of (1S,2S)-U50,488, (1S,2R)-U50,488, or (1R,2R) U50,488 on colon sensory neurons in vitro decreased voltage-activated sodium currents. This inhibition by U50,488 and its stereoisomers was not opioid receptor-mediated because it could not be reversed by the opioid receptor antagonist naloxone and was also not a G protein-mediated effect. The results reported here suggest that the visceral antinociceptive effects of U50,488 and its stereoisomers are contributed to by their peripheral sodium channel blocking actions. PMID- 11877502 TI - Responses of superficial dorsal horn neurons to intradermal serotonin and other irritants: comparison with scratching behavior. AB - Scratching behavior is used to assess itch sensation in animals, but few studies have addressed the relative scratch-inducing capacity of different algesic and pruritic chemicals. Furthermore, central neural mechanisms underlying itch are not well understood. We used electrophysiological and behavioral methods to investigate the ability of several irritant chemicals to excite neurons in the superficial dorsal horn, as well as to elicit scratching, in rats. In anesthetized rats, single neurons in the superficial lumbar dorsal horn, identified by their responsiveness to intracutaneous (ic) histamine, were classified as wide dynamic range (WDR) or nociceptive-specific (NS). Serotonin (5 HT) given ic to the paw excited most (88%) WDR and NS neurons over a prolonged time course (often up to 40 min). 5-HT-evoked responses exhibited significant tachyphylaxis. Most neurons also gave shorter-duration responses to ic capsaicin (92%) and mustard oil (71%). In separate behavioral experiments, significant dose related hind limb scratching directed at the ic injection site in the back of the neck was elicited by 5-HT over a time course similar to that of evoked neuronal firing. A second 5-HT injection made 40 min later at the same site elicited significantly less scratching. Formalin also elicited scratching that was not dose-related and less than that evoked by 5-HT. 5-HT and Formalin also evoked head or whole-body shakes that were significantly correlated with scratching. Neither histamine, capsaicin, nor vehicle controls elicited significant scratching or shaking. In rats, 5-HT appears to be more pruritogenic than histamine as assessed by scratching and shaking behavior, and excites superficial dorsal horn neurons over a behaviorally relevant time course. However, because most neurons additionally responded to pain-producing stimuli, they are not itch specific. They might nonetheless contribute to neural pathways that distinguish between pain and itch based on some neural mechanism such as frequency coding. PMID- 11877503 TI - Spinal inhibitory effects of cardiopulmonary afferent inputs in monkeys: neuronal processing in high cervical segments. AB - Noxious stimulation of spinal afferents inhibits primate spinothalamic tract (STT) neurons in segments distant from the region of afferent entry. Inhibitory effects of cardiopulmonary sympathetic afferent (CPSA) stimulation remain after C(1) transection but disappear with spinal transection between C(3) and C(7). We hypothesized that spinal inhibitory effects produced by CPSA stimulation are processed by neurons in C(1)-C(3) segments. One purpose of this study in anesthetized monkeys was to determine whether chemical activation of high cervical neurons reduced sacral STT cell responses to colorectal distension (CRD) and urinary bladder distension (UBD). First, effects and interactions of pelvic and cardiopulmonary visceral afferent inputs were determined in 10 monkeys on extracellular activity of sacral STT neurons recorded in deep dorsal horn. CRD and UBD increased activity in 95 and 91% of sacral STT neurons, respectively. CPSA and cardiopulmonary vagal stimulation decreased activity in 84 and 56% of STT neurons, respectively. CPSA stimulation decreased CRD-evoked activity in six of eight sacral STT neurons and decreased UBD-evoked activity in five of eight STT neurons tested. Excitatory amino acid application at C2 segment decreased CRD evoked responses in 7 of 10 sacral STT neurons and decreased UBD-evoked responses in 9 of 12 STT neurons. The second purpose of this study was to examine responses of C(1)-C(3) descending propriospinal neurons to stimulation of cardiopulmonary afferent fibers. If C(1)-C(3) neurons process CPSA input to suppress STT transmission, then CPSA stimulation should excite C(1)-C(3) neurons with descending projections. Effects of thoracic vagus nerve stimulation also were examined. Vagal stimulation inhibits STT neurons in segments below C(3) but excites C(1)-C(3) STT neurons; we theorized that vagal inhibition of sensory transmission might relay in high cervical segments and, therefore, excite C(1) C(3) descending propriospinal neurons. Extracellular discharge rate was recorded for C(1)-C(3) neurons antidromically activated from thoracic or lumbar spinal cord in 24 monkeys. CPSA stimulation increased activity of 16 of 45 neurons and inhibited one cell. Thoracic vagus stimulation increased activity of 20 of 43 neurons and inhibited one cell; stimulation of abdominal vagus fibers did not affect activity of six of six cells that were excited by thoracic vagal input. Mechanical stimulation of somatic fields excited 30 of 41 neurons tested. All neurons activated by visceral input received convergent somatic input from noxious pinch of somatic receptive fields that generally included the neck and upper body; 11 C(1)-C(3) propriospinal neurons did not respond to any afferent input examined. Results of these studies were consistent with the idea that modulation of spinal nociceptive transmission might involve neuronal connections in high cervical segments. PMID- 11877504 TI - Resilient RTN fast spiking in Kv3.1 null mice suggests redundancy in the action potential repolarization mechanism. AB - Fast spiking (FS), GABAergic neurons of the reticular thalamic nucleus (RTN) are capable of firing high-frequency trains of brief action potentials, with little adaptation. Studies in recombinant systems have shown that high-voltage-activated K(+) channels containing the Kv3.1 and/or Kv3.2 subunits display biophysical properties that may contribute to the FS phenotype. Given that RTN expresses high levels of Kv3.1, with little or no Kv3.2, we tested whether this subunit was required for the fast action potential repolarization mechanism essential to the FS phenotype. Single- and multiple-action potentials were recorded using whole cell current clamp in RTN neurons from brain slices of wild-type and Kv3.1 deficient mice. At 23 degrees C, action potentials recorded from homozygous Kv3.1 deficient mice (Kv3.1(-/-)) compared with their wild-type (Kv3.1(+/+)) counterparts had reduced amplitudes (-6%) and fast after-hyperpolarizations ( 16%). At 34 degrees C, action potentials in Kv3.1(-/-) mice had increased duration (21%) due to a reduced rate of repolarization (-30%) when compared with wild-type controls. Action potential trains in Kv3.1(-/-) were associated with a significantly greater spike decrement and broadening and a diminished firing frequency versus injected current relationship (F/I) at 34 degrees C. There was no change in either spike count or maximum instantaneous frequency during low threshold Ca(2+) bursts in Kv3.1(-/-) RTN neurons at either temperature tested. Our findings show that Kv3.1 is not solely responsible for fast spikes or high frequency firing in RTN neurons. This suggests genetic redundancy in the system, possibly in the form of other Kv3 members, which may suffice to maintain the FS phenotype in RTN neurons in the absence of Kv3.1. PMID- 11877505 TI - Age-dependent effects of peripheral inflammation on the electrophysiological properties of neonatal rat dorsal horn neurons. AB - The aim of this study was to investigate the postnatal development of spinal cord neurophysiological mechanisms of inflammatory pain. The effect of hindpaw inflammation on the properties of neonatal spinal dorsal horn cells was investigated in urethane-anesthetized newborn rats using in vivo single-unit extracellular recordings. Responses to cutaneous mechanical and electrical A and C fiber stimulation were recorded at postnatal day (P) 3, 10, and 21 in pups that had received a unilateral intraplantar carageenan injection (1%, 1 microl/g body wt) 2-5 h earlier and compared with age-matched controls. At all three ages, carageenan inflammation increased A fiber evoked sensitization, spontaneous activity, and the suprathreshold response magnitude of dorsal horn cells. Receptive field size, which normally decreases with postnatal age, was unaffected by inflammation in P3 and P10 pups but significantly increased at P21 so that the size distribution closely resembled that in control P3 pups. Mechanical thresholds of individual dorsal horn neurons were not altered by carageenan inflammation at any age. The results show that some dorsal horn cell properties that are likely to underlie inflammatory hypersensitivity such as increased spontaneous activity and response magnitude are observed from the earliest postnatal age examined (P3). However inflammation induced expansion of mechanical receptive field size is not observed until at least the second postnatal week. These results have implications for the postnatal processing of inflammatory pain. PMID- 11877506 TI - Development and serotonergic modulation of NMDA bursting in rat trigeminal motoneurons. AB - The development of N-methyl-D-aspartate (NMDA)-induced burst discharge in rat trigeminal motoneurons (TMNs) between postnatal days P1 and P10 was examined using whole cell patch-clamp recording methods in brain slices. Bath application of NMDA (50 microM) induced a Mg(2+)-dependent rhythmical bursting activity starting around P8. Prior to the onset of bursting, the membrane potential depolarized and the input resistance increased. Hyperpolarization of the membrane potential with extrinsic current demonstrated a narrow window of membrane potential where maintained rhythmical burst discharge was evident. In P1-P4 neurons, NMDA application produced membrane depolarization and a minimal change in input resistance, but no burst activity at any membrane potential. Voltage clamp analysis indicated that the bursting activity was related to the presence or absence of a voltage-dependent Mg(2+) block and induction of a negative slope conductance (NSC) region in the I(NMDA)-V relationship. Regardless of age, reduction of extracellular Mg(2+) from 1 mM to 30 microM enhanced I(NMDA) at voltages negative to -60 mV. However, in 1 mM Mg(2+), P1-P4 neurons were devoid of a prominent NSC region compared with P8-P10 neurons, suggesting that the efficacy of depolarization in unblocking the NMDA receptors increased with age. NMDA bursting was not dependent on calcium influx through voltage-gated calcium channels (VGCC) but did require a minimal concentration of Ca(2+) in the bath. Intracellular bis-(o-aminophenoxy)-N,N,N',N'-tetraacetic acid application suppressed burst discharge completely, suggesting that intracellular Ca(2+) directly, or via second-messenger systems, regulates NMDA receptor activity and bursting. Interestingly, NMDA bursting could be induced in P1-P4 neurons by simultaneous bath application of serotonin (5-HT, 10 microM), which by itself did not produce bursting, suggesting an "enabling" role for 5-HT. Voltage-clamp analysis demonstrated that the NMDA/5-HT bursting resulted from induction of an NSC in the I-V relationship of total membrane current. 5-HT by itself produced no such effect. The mechanisms for this effect were due to an enhancement of the NSC region of the I(NMDA)-V relationship and reduction of a presumed leak current by 5-HT. These data indicate that NMDA bursting in trigeminal motoneurons is developmentally regulated and subject to neuromessenger modulation. Control of the Mg(2+) sensitivity of the NMDA receptor and voltage-dependent block by neuromessengers could be an effective means to control the efficacy of glutamatergic synaptic drive to motoneurons during rhythmical oral-motor activity at early postnatal ages. PMID- 11877507 TI - Motor facilitation while observing hand actions: specificity of the effect and role of observer's orientation. AB - Action observation enhances cortico-spinal excitability. Here we tested the specificity of this effect and the role played by the orientation of the observer. Ten normal subjects observed video clips of right hand performing three different finger movements (thumb ab-/adduction, index ab-/adduction, index extens-/flexion) in two different orientations (Away, i.e., natural hand orientation facing out from the observer; or Toward, i.e., unnatural hand orientation facing toward the observer). Motor-evoked potentials (MEPs) induced by transcranial magnetic stimulation (TMS) were recorded from the abductor pollicis brevis (APB) and the first dorsal interosseus (FDI) muscles. Movement direction of the index finger was recorded using force transducers. Facilitation of MEP size was significantly greater for APB during observation of thumb movements and for FDI during observation of index finger movements. Facilitation of MEP size was significantly greater when the hand presented on screen was facing out from and corresponding to that of the observer (Away orientation). The direction of the index finger movement evoked by TMS shifted toward extension/flexion versus ab-/adduction matching the observed movement. Our results give further evidence that observation of a movement enhances motor output to the muscles involved in the movement and facilitates the observed action. In addition, we provide novel evidence about the high degree of specificity of this observation-induced motor cortical modulation. The degree of modulation depends on hand orientation. The modulation is maximal when the observed action corresponds to the orientation of the observer. PMID- 11877508 TI - Cerebellar damage impairs automaticity of a recently practiced movement. AB - It has been suggested that the cerebellum plays a critical role in learning to make movements more "automatic" (i.e., requiring less attention to the details of a movement). We hypothesized that cerebellar damage compromises learning of movement automaticity, resulting in increased attentional demands for movement control. The purpose of our study was to determine whether cerebellar damage disrupts the ability to make a practiced movement more automatic. We developed a dual task paradigm using two tasks that did not have overlapping sensory or motor requirements for execution. Our motor task required subjects to maintain an upright posture while performing a figure-8 movement using their arm. This motor task was chosen to simulate requirements of everyday movements (e.g., standing while reaching for objects), but it was novel enough to require practice for improvement. Our secondary task was an auditory vigilance task where subjects listened to letter sequences and were asked to identify the number of times a target letter was heard. We tested controls and people with cerebellar damage as they practiced the movement task alone and then performed it with the auditory task. We recorded 3D position data from the arm, trunk, and leg during the movement task. Errors were recorded for both the movement and the letter tasks. Our results show that cerebellar subjects can improve the movement to a very limited extent with practice. Unlike controls, the motor performance of cerebellar subjects deteriorates to prepractice levels when attention is focused away from the movement during dual task trials. Control subjects' insensitivity to dual task interference after practice was due to learned movement automaticity and was not a reflection of better dual task performance generally. Overall, our findings suggest that the cerebellum may be important for shifting movement performance from an attentionally demanding (unpracticed) state to a more automatic (practiced) state. PMID- 11877510 TI - Simulation of different firing patterns in paired spider mechanoreceptor neurons: the role of Na(+) channel inactivation. AB - The spider VS-3 slit-sense organ contains two types of primary mechanoreceptor neurons that are morphologically similar but have different electrical behavior. Type A neurons fire only one or two action potentials in response to a mechanical or electrical step of any amplitude above the threshold, whereas type B neurons fire prolonged bursts of action potentials in response to similar stimuli. Voltage-clamp studies have shown that two voltage-activated ion currents, a noninactivating potassium current and an inactivating sodium current, dominate the firing behavior. We simulated the electrical behavior of the two neuron types, using a simplified form of Hodgkin-Huxley model based on published voltage clamp and current-clamp recordings. Changing only two parameters of sodium inactivation, the slope of the h(infinity) curve and the time constant of recovery from inactivation, allowed a complete switch between the two firing patterns. Our simulations support previous evidence that sodium inactivation controls the firing properties of these neurons and indicate that two parameter changes are needed to achieve complete transformation between the two neuron types. PMID- 11877511 TI - Increase in adenosine sensitivity in the nucleus accumbens following chronic morphine treatment. AB - There is a growing body of evidence suggesting that the neuromodulator adenosine is involved in drug addiction and withdrawal and that adenosine signaling pathways may offer new targets for therapeutic treatments of addiction. Recent studies have suggested that chronic exposure to drugs of abuse may alter adenosine metabolism in the nucleus accumbens, a brain region critically involved in drug addiction and withdrawal. The present study examined the effects of chronic morphine treatment on the ability of adenosine to inhibit excitatory postsynaptic currents in nucleus accumbens medium spiny neurons. It was found that chronic morphine treatment via subcutaneous implantation of morphine pellets in rats for 1 wk did not alter the level of adenosine-mediated tonic inhibition of nucleus accumbens excitatory synapses. However, chronic morphine treatment did induce a leftward shift in the adenosine dose-response curve, indicating an increase in the sensitivity of synaptic currents to exogenously applied adenosine. This shift was not due to a change in adenosine receptors or their effectors, because chronic morphine treatment had no effect on the dose-response relationship of a nonmetabolized adenosine receptor agonist. When adenosine transport was blocked, the ability of chronic morphine to shift the adenosine dose-response curve was eliminated. These experiments suggest that the increase in the sensitivity of nucleus accumbens synapses to the inhibitory effects of adenosine may be due to a decrease in adenosine transport. The identification of these changes in the adenosine system after chronic drug exposure may help identify new therapeutic strategies aimed at easing withdrawal from opioids. PMID- 11877509 TI - Regulation of the timing and pattern of action potential generation in rat subthalamic neurons in vitro by GABA-A IPSPs. AB - The regulation of activity in the subthalamic nucleus (STN) by GABAergic inhibition from the reciprocally connected globus pallidus (GP) plays an important role in normal movement and disorders of movement. To determine the precise manner in which GABAergic synaptic input, acting at A-type receptors, influences the firing of STN neurons, we recorded the response of STN neurons to GABA-A inhibitory postsynaptic potentials (IPSPs) that were evoked by supramaximal electrical stimulation of the internal capsule using the perforated patch technique in slices at 37 degrees C. The mean equilibrium potential of the GABA-A IPSP (EGABA-A IPSP) was -79.4 +/- 7.0 mV. Single IPSPs disrupted the spontaneous oscillation that underlies rhythmic single-spike firing in STN neurons. As the magnitude of IPSPs increased, the effectiveness of prolonging the interspike interval was related more strongly to the phase of the oscillation at which the IPSP was evoked. Thus the largest IPSPs tended to reset the oscillatory cycle, whereas the smallest IPSPs tended to produce relatively phase-independent delays in firing. Multiple IPSPs were evoked at various frequencies and over different periods and their impact was studied on STN neurons held at different levels of polarization. Multiple IPSPs reduced and/or prevented action potential generation and/or produced sufficient hyperpolarization to activate a rebound depolarization, which generated a single spike or restored rhythmic spiking and/or generated a burst of activity. The pattern of IPSPs and the level of polarization of STN neurons were critical in determining the nature of the response. The duration of bursts varied from 20 ms to several hundred milliseconds, depending on the intrinsic rebound properties of the postsynaptic neuron. These data demonstrate that inhibitory input from the GP can produce a range of firing patterns in STN neurons, depending on the number and frequencies of IPSPs and the membrane properties and voltage of the postsynaptic neuron. PMID- 11877512 TI - Effects of K+ channel blockers on developing rat myelinated CNS axons: identification of four types of K+ channels. AB - Four blockers of voltage-gated potassium channels (Kv channels) were tested on the compound action potentials (CAPs) of rat optic nerves in an attempt to determine the regulation of Kv channel expression during the process of myelination. Before myelination occurred, 4-aminopyridine (4-AP) increased the amplitude, duration, and refractory period of the CAPs. On the basis of their pharmacological sensitivity, 4-AP-sensitive channels were divided in two groups, the one sensitive to kaliotoxin (KTX), dendrotoxin-I (DTX-I), and 4-AP, and the other sensitive only to 4-AP. In addition, tetraethylammonium chloride (TEA) applied alone broadened the CAPs. At the onset of myelination, DTX-I induced a more pronounced effect than KTX; this indicates that a fourth group of channels sensitive to 4-AP and DTX-I but insensitive to KTX had developed. The effects of KTX and DTX-I gradually disappeared during the period of myelination. Electron microscope findings showed that the disappearance of these effects was correlated with the ongoing process of myelination. This was confirmed by the fact that DTX I and KTX enlarged the CAPs of demyelinated adult optic nerves. These results show that KTX- and DTX-sensitive channels are sequestrated in paranodal regions. During the process of myelination, KTX had less pronounced effects than DTX-I on demyelinated nerves, which suggests that the density of the KTX-sensitive channels decreased during this process. By contrast, 4-AP increased the amplitude, duration, and refractory period of the CAPs at all the ages tested and to a greater extent than KTX and DTX-I. The effects of TEA alone also gradually disappeared during this period. However, effects of TEA on CAPs were observed when this substance was applied after 4-AP to the adult optic nerve; this shows that TEA-sensitive channels are not masked by the myelin sheath. In conclusion, the process of myelination seems to play an important part in the regulation and setting of Kv channels in optic nerve axons. PMID- 11877513 TI - Nociceptive quality of the laser-evoked blink reflex in humans. AB - Laser radiant-heat pulses selectively excite the free nerve endings in the superficial layers of the skin and activate mechano-thermal nociceptive afferents; when directed to the perioral or supraorbital skin, high-intensity laser pulses evoke a blink-like response in the orbicularis oculi muscle (the laser blink reflex, LBR). We investigated the functional properties (startle or nociceptive origin) of the LBR and sought to characterize its central pathways. Using high-intensity CO(2)-laser stimulation of the perioral or supraorbital regions and electromyographic (EMG) recordings from the orbicularis oculi muscles, we did five experiments in 20 healthy volunteers. First, to investigate whether the LBR is a startle response, we studied its habituation to expected rhythmic stimuli and to unexpected arrhythmic stimuli. To assess its possible nociceptive quality, we studied changes in the LBR and the R2 component of the electrical blink reflex after a lidocaine-induced supraorbital nerve block and after intramuscular injection of the opiate fentanyl and the opiate-antagonist naloxone. To characterize the central pathways for the LBR, we investigated the interaction between the LBR and the three components of the blink reflex (R1, R2, and R3) by delivering laser pulses to the perioral or supraorbital regions before or after electrical stimulation of the supraorbital nerve at various interstimulus intervals. Finally, to gain further information on the central LBR pathways, using two identical CO(2)-laser stimulators, we studied the LBR recovery curves with paired laser pulses delivered to adjacent forehead points at interstimulus intervals from 250 ms to 1.5 s. The LBR withstood relatively high frequency rhythmic stimulations, and unexpected laser pulses failed to evoke larger responses. When lidocaine began to induce hypoalgesia (about 5 min after the injection), the LBR was abolished, whereas R2 was only partly suppressed 10 min after the injection. Fentanyl injection induced strong, naloxone-reversible, LBR suppression (the response decreased to 25.3% of predrug values at 10 min and to 4% at 20 min), whereas R2 remained appreciably unchanged. Whether directed to the perioral or supraorbital regions, preceding laser pulses strongly suppressed R2 and R3 though not R1. Conversely, preceding electrical stimuli to the supraorbital nerve suppressed the LBR. In response to paired stimuli, the LBR recovered significantly faster than R2. These findings indicate that the LBR is a nociceptive reflex, which shares part of the interneuron chain mediating the nonnociceptive R2 blink reflex, probably in the medullary reticular formation. The LBR may prove useful for studying the pathophysiology of orofacial pain syndromes. PMID- 11877515 TI - GABA application to hippocampal CA3 or CA1 stratum lacunosum-moleculare excites an interneuron network. AB - Whole cell voltage-clamp recording and focal application of the neurotransmitter gamma-aminobutyric acid (GABA) were used to investigate the ability of exogenous GABA applied to different locations within the guinea pig hippocampal slice to trigger a giant GABA-mediated postsynaptic current (GPSC) in pyramidal cells. A GPSC reflects the synchronous release of GABA from a group of interneurons. Recordings were done in the presence of 4-aminopyridine (4-AP) and blockers of ionotropic glutamatergic synaptic transmission. Spontaneous GPSCs occurred rhythmically in pyramidal cells under these conditions. Brief focal pressure application of GABA (500 microM; 30-200 ms) to CA3 stratum lacunosum-moleculare (SLM) or to the border between CA3 s. radiatum (SR) and SLM triggered an "all-or none" GPSC in CA3 and CA1 pyramidal cells that looked like the spontaneous GPSCs. During the refractory period following a spontaneous GPSC, application of GABA could not trigger a GPSC. Both spontaneous GPSCs and GPSCs triggered by exogenous GABA were blocked by suppressing synaptic transmission with high Mg(2+)/low Ca(2+) bath solution. On the other hand, focal application of GABA to CA3 s. oriens (SO) or to proximal SR did not trigger a GPSC in the CA3 pyramidal cell; instead it produced a graded response. Focal application of GABA to regions other than CA3 was also tested. Focal application of GABA to CA1 SLM always triggered a GPSC in the CA3 pyramidal cell. Focal application of GABA within the outer two thirds of the dentate molecular layer often elicited a GPSC in the CA3 pyramidal cell. In contrast, focal application of GABA to CA1 SO, to CA1 SR, or to the hilus elicited no current response in the CA3 pyramidal cell. These data indicate that the GPSC recorded in pyramidal cells that was triggered by focal GABA application resulted from the synchronous synaptic release of GABA from activated interneurons rather than from the binding of exogenous GABA to receptors on the pyramidal cell. Furthermore, the "all-or-none" nature of the response to SLM GABA applications of different durations indicates that the exogenous GABA was exciting (directly or indirectly) some members of a network of interneurons, which in turn recruited the rest of the network, rather than individually activating each interneuron that contributed to the GPSC. Interestingly, the effective sites of GABA application--CA3 SLM, CA1 SLM, and the outer two-thirds of the dentate molecular layer--are also the sites which receive direct innervation from the entorhinal cortex in an intact animal. PMID- 11877514 TI - Postsynaptic induction and presynaptic expression of group 1 mGluR-dependent LTD in the hippocampal CA1 region. AB - Activation of metabotropic glutamate receptors (mGluRs) with the group I mGluR selective agonist (R,S)-3,5-dihydroxyphenylglycine (DHPG) induces a long-term depression (LTD) of excitatory synaptic transmission in the CA1 region of the hippocampus. Here we investigated the potential roles of pre- and postsynaptic processes in the DHPG-induced LTD at excitatory synapses onto hippocampal pyramidal cells in the mouse hippocampus. Activation of mGluRs with DHPG, but not ACPD, induced LTD at both Schaffer collateral/commissural fiber synapses onto CA1 pyramidal cells and at associational/commissural fiber synapses onto CA3 pyramidal cells. DHPG-induced LTD was blocked when the G-protein inhibitor guanosine-5'-O-(2-thiodiphosphate) was selectively delivered into postsynaptic CA1 pyramidal cells via an intracellular recording electrode, suggesting that DHPG depresses synaptic transmission through a postsynaptic, GTP-dependent signaling pathway. The effects of DHPG were also strongly modulated, however, by experimental manipulations that altered presynaptic calcium influx. In these experiments, we found that elevating extracellular Ca(2+) concentrations ([Ca(2+)](o)) to 6 mM almost completely blocked the effects of DHPG, whereas lowering [Ca(2+)](o) to 1 mM significantly enhanced the ability of DHPG to depress synaptic transmission. Enhancing Ca(2+) influx by prolonging action potential duration with bath applications of the K(+) channel blocker 4 aminopyridine (4-AP) also strongly reduced the effects of DHPG in the presence of normal [Ca(2+)](o) (2 mM). Although these findings indicate that alterations in Ca(2+)-dependent signaling processes strongly regulate the effects of DHPG on synaptic transmission, they do not distinguish between potential pre- versus postsynaptic sites of action. We found, however, that while inhibiting both pre- and postsynaptic K(+) channels with bath-applied 4-AP blocked the effects of DHPG; inhibition of postsynaptic K(+) channels alone with intracellular Cs(+) and TEA had no effect on the ability of DHPG to inhibit synaptic transmission. This suggests that presynaptic changes in transmitter release contribute to the depression of synaptic transmission by DHPG. Consistent with this, DHPG induced a persistent depression of both AMPA and N-methyl-D-aspartate receptor-mediated components of excitatory postsynaptic currents in voltage-clamped pyramidal cells. Together our results suggest that activation of postsynaptic mGluRs suppresses transmission at excitatory synapses onto CA1 pyramidal cells through presynaptic effects on transmitter release. PMID- 11877516 TI - Changes in calcium signaling during postembryonic dendritic growth in Manduca sexta. AB - Activity-dependent Ca(2+) influx plays crucial roles in adult and developing nervous systems through its influence on signal processing, synaptic plasticity, and neuronal differentiation. The responses to internal Ca(2+) elevations vary depending on the spatial distribution of Ca(2+) accumulation in different cell compartments. In this study, the mechanisms and the distribution of Ca(2+) accumulation are addressed by in situ Ca(2+) imaging of an identified insect motoneuron, MN5, at critical stages of postembryonic life. During metamorphosis of Manduca sexta, MN5 undergoes extensive dendritic regression followed by regrowth. The time course, amplitude, and distribution of Ca(2+) accumulation within MN5 change during development. During the initial stage of rapid dendritic growth and branching, dendritic growth cones are present, and voltage-dependent Ca(2+) currents are small. At this stage, activity-induced elevations of internal Ca(2+) are largest in the distal dendrites, suggesting that the density of voltage-gated Ca(2+) channels is highest in these regions. Later phases of dendritic growth are accompanied by the transient occurrence of prominent Ca(2+) spikes. Single Ca(2+) spikes cause robust Ca(2+) influx of similar amplitudes and time courses in all central compartments of MN5. The resting Ca(2+) levels also increase during development. Ca(2+)-induced Ca(2+) release from intracellular stores did not contribute to the elevations measured at either stage, although Ca(2+) stores are present in the dendrites. These developmental changes of the internal Ca(2+) signaling are consistent with a regulatory role for activity dependent Ca(2+) influx in postembryonic dendritic growth. PMID- 11877517 TI - Mitochondrial Ca(2+) buffering regulates synaptic transmission between retinal amacrine cells. AB - The diverse functions of retinal amacrine cells are reliant on the physiological properties of their synapses. Here we examine the role of mitochondria as Ca(2+) buffering organelles in synaptic transmission between GABAergic amacrine cells. We used the protonophore p-trifluoromethoxy-phenylhydrazone (FCCP) to dissipate the membrane potential across the inner mitochondrial membrane that normally sustains the activity of the mitochondrial Ca(2+) uniporter. Measurements of cytosolic Ca(2+) levels reveal that prolonged depolarization-induced Ca(2+) elevations measured at the cell body are altered by inhibition of mitochondrial Ca(2+) uptake. Furthermore, an analysis of the ratio of Ca(2+) efflux on the plasma membrane Na-Ca exchanger to influx through Ca(2+) channels during voltage steps indicates that mitochondria can also buffer Ca(2+) loads induced by relatively brief stimuli. Importantly, we also demonstrate that mitochondrial Ca(2+) uptake operates at rest to help maintain low cytosolic Ca(2+) levels. This aspect of mitochondrial Ca(2+) buffering suggests that in amacrine cells, the normal function of Ca(2+)-dependent mechanisms would be contingent upon ongoing mitochondrial Ca(2+) uptake. To test the role of mitochondrial Ca(2+) buffering at amacrine cell synapses, we record from amacrine cells receiving GABAergic synaptic input. The Ca(2+) elevations produced by inhibition of mitochondrial Ca(2+) uptake are localized and sufficient in magnitude to stimulate exocytosis, indicating that mitochondria help to maintain low levels of exocytosis at rest. However, we found that inhibition of mitochondrial Ca(2+) uptake during evoked synaptic transmission results in a reduction in the charge transferred at the synapse. Recordings from isolated amacrine cells reveal that this is most likely due to the increase in the inactivation of presynaptic Ca(2+) channels observed in the absence of mitochondrial Ca(2+) buffering. These results demonstrate that mitochondrial Ca(2+) buffering plays a critical role in the function of amacrine cell synapses. PMID- 11877518 TI - Divergent movement of adjacent whiskers. AB - The current view of whisker movement is that approximately 25 whiskers on each side of the face move in synchrony. To determine whether whiskers are constrained to move together, we trained rats to use two whiskers on the same side of the face in simple behavioral tasks and videotaped the whiskers during the task. Here we report that the movement of adjacent whiskers is usually synchronous but can diverge: 1) the distance between whiskers can vary dramatically during movement; 2) one whisker can move while the second one remains stationary; 3) two whiskers can simultaneously move in opposite directions; and 4) one whisker can be maintained in contact with an object while the other is retracted and protracted. The frequency of whisker movement during the task falls within the previously reported range for rats whisking freely into air or performing roughness discrimination with their whiskers. Our data also suggest that whisker movement can be divided into three distinct phases: protraction, retraction, and a measurable delay between these movements. We conclude that, although whiskers often move in concert, adjacent caudal whiskers can be moved independently of each other. PMID- 11877519 TI - Calcium transients in the garter snake vomeronasal organ. AB - The signaling cascade involved in chemosensory transduction in the VN organ is incompletely understood. In snakes, the response to nonvolatile prey chemicals is mediated by the vomeronasal (VN) system. Using optical techniques and fluorescent Ca(2+) indicators, we found that prey-derived chemoattractants produce initially a transient cytosolic accumulation of [Ca(2+)](i) in the dendritic regions of VN neurons via two pathways: Ca(2+) release from IP(3)-sensitive intracellular stores and, to a lesser extent, Ca(2+) influx through the plasma membrane. Both components seem to be dependent on IP(3) production. Chemoattractants evoke a short-latency Ca(2+) elevation even in the absence of extracellular Ca(2+), suggesting that in snake VN neurons, Ca(2+) release from intracellular stores is independent of a preceding Ca(2+) influx, and both components are activated in parallel during early stages of chemosensory transduction. Once the response develops in apical dendritic segments, other mechanisms can also contribute to the amplification and modulation of these chemoattractant-mediated cytosolic Ca(2+) transients. In regions close to the cell bodies of the VN neurons, the activation of voltage-sensitive Ca(2+) channels and a Ca(2+)-induced Ca(2+) release from intracellular ryanodine-sensitive stores secondarily boost initial cytosolic Ca(2+) elevations increasing their magnitude and durations. Return of intracellular Ca(2+) to prestimulation levels appears to involve a Ca(2+) extrusion mediated by a Na(+)/Ca(2+) exchanger mechanism that probably plays an important role in limiting the magnitude and duration of the stimulation-induced Ca(2+) transients. PMID- 11877520 TI - Contribution of primate magnocellular red nucleus to timing of hand preshaping during reaching to grasp. AB - Magnocellular red nucleus (RNm) is involved in controlling goal-directed limb movements such as reaching to grasp. We tested two hypotheses related to RNm's role in controlling reach-to-grasp movements. One hypothesis is that forelimb RNm neurons are grasp specific, and the other is that they specify the timing of metacarpi-phalangeal (MCP) extension to preshape the hand during the appropriate phase of the reach. We recorded single-unit discharge while monkeys performed two behavioral tasks that elicited similar reaches but differed in grasp. One task consisted of a reach with a precision grasp that elicited independent use of thumb and forefinger; the other included a whole-hand grasp that elicited concerted use of the four fingers. Most RNm neurons tested were engaged strongly during both the whole-hand and precision tasks, and the magnitude of discharge modulation did not differ between tasks. Thus most RNm neurons are not grasp specific but, instead, may contribute to behavioral features common to the two tasks. Two methods were used to investigate relations between single-unit discharge and kinematic data from the same individual trials of the whole-hand and precision tasks for a subset of forelimb RNm neurons. One method focused on correlations between parameters of RNm discharge and the duration, amplitude, and velocity of rotation of forelimb joints for each of the tasks. The second method compared between-task differences in times of peak neuronal discharge to between task differences in times of rotations of forelimb joints. Parameters of reach related RNm discharge were more frequently correlated with parameters of MCP extension than with parameters of rotation of wrist, elbow, and shoulder joints. Analyses of temporal relations between discharge and kinematic data during both the whole-hand and precision tasks indicate that discharge was time locked most frequently to MCP extension and, to a lesser extent, elbow extension during both tasks. We conclude that RNm may command muscle synergies that provide a basic preshape of the hand at the appropriate phase of limb transport. In addition, the timing of RNm's contribution to hand preshaping varies with the behavioral requirements of the task. PMID- 11877521 TI - Influence of reward expectation on visuospatial processing in macaque lateral prefrontal cortex. AB - The lateral prefrontal cortex (LPFC) has been implicated in visuospatial processing, especially when it is required to hold spatial information during a delay period. It has also been reported that the LPFC receives information about expected reward outcome. However, the interaction between visuospatial processing and reward processing is still unclear because the two types of processing could not be dissociated in conventional delayed response tasks. To examine this, we used a memory-guided saccade task with an asymmetric reward schedule and recorded 228 LPFC neurons. The position of the target cue indicated the spatial location for the following saccade and the color of the target cue indicated the reward outcome for a correct saccade. Activity of LPFC was classified into three main types: S-type activity carried only spatial signals, R-type activity carried only reward signals, and SR-type activity carried both. Therefore only SR-type cells were potentially involved in both visuospatial processing and reward processing. SR-type activity was enhanced (SR+) or depressed (SR-) by the reward expectation. The spatial discriminability as expressed by the transmitted information was improved by reward expectation in SR+ type. In contrast, when reward information was coded by an increase of activity in the reward-absent condition (SR- type), it did not improve the spatial representation. This activity appeared to be involved in gaze fixation. These results extend previous findings suggesting that the LPFC exerts dual influences based on predicted reward outcome: improvement of memory-guided saccades (when reward is expected) and suppression of inappropriate behavior (when reward is not expected). PMID- 11877522 TI - High-speed VSD imaging of visually evoked cortical waves: decomposition into intra- and intercortical wave motions. AB - In the pond turtle, Pseudemys scripta elegans, visually evoked cortical waves propagate at different velocities within the primary visual area compared with waves that pass into the secondary visual area. In an effort to separate intra- and intercortical wave motions, movies of visually evoked cortical waves recorded by high-speed voltage-sensitive dye (VSD) imaging were subjected to Karhunen Loeve (KL) decomposition. This procedure decomposes the VSD movies into a series of basis images that capture different spatial patterns of coherent activity. Most of the energy of the compound wave motion (>95%) was captured by the three largest basis images, M(1,1), M(1,2), and M(2,1). Based on visual comparison with maps of wave front latency, KL basis image M(1,2) appears to capture the spread of depolarization within the primary visual area, whereas KL basis image M(2,1) appears to capture the spread of depolarization from the primary into the secondary visual area. The contribution of different basis images to the intra- and intercortical wave motions was tested by reconstructing the response using different combinations of KL basis images. Only KL basis images M(1,1) and M(1,2) were needed to reconstruct intracortical wave motion, while basis images M(1,1) and M(2,1) were needed to reconstruct intercortical wave motion. It was also found that the direction and speed of wave propagation could be deduced by visual inspection of the basis image projections on to the original data set. The relative advantage of KL decomposition for the analysis of complex wave motions captured by VSD imaging is discussed. PMID- 11877523 TI - Pharmacological characterization of glycine-gated chloride currents recorded in rat hippocampal slices. AB - An inhibitory role for strychnine-sensitive glycine-gated chloride channels (GlyRs) in mature hippocampus has been overlooked, largely due to the misconception that GlyR expression ceases early during development and to few functional studies demonstrating their presence. As a result, little is known regarding the physiological and pharmacological properties of native GlyRs expressed by hippocampal neurons. In this study, we used pharmacological tools and whole cell patch-clamp recordings of CA1 pyramidal cells and interneurons in acutely prepared hippocampal slices from 3- to 4-wk old rats to characterize these understudied receptors. We show that glycine application to recorded pyramidal cells and interneurons elicited strychnine-sensitive chloride-mediated currents (I(gly)) that did not completely desensitize in the continued presence of agonist but reached a steady state at 45-60% of the peak amplitude. Additionally, the inhibitory amino acid, taurine, which has been shown to activate GlyRs in other systems, activated GlyRs expressed by both pyramidal cells and interneurons, although with much less potency than glycine, having an EC(50) 10-fold higher. To examine the potential subunit composition of hippocampal GlyRs, we tested the effect of the GABA(A) receptor antagonist, picrotoxin, on I(gly) recorded from both cell types. At low micromolar concentrations of picrotoxin (< or =100 microM), which selectively block alpha homomeric GlyRs, I(gly) was partially attenuated in both cell types, indicating that alpha homomeric receptors are expressed by pyramidal cells and interneurons. At picrotoxin concentrations < or =1 mM, approximately 10-20% of the whole cell current remained, suggesting that alphabeta heteromeric GlyRs are also expressed because this subtype of GlyR is relatively resistant to picrotoxin antagonism. Finally, we examined whether hippocampal GlyRs are modulated by zinc. Consistent with previous reports in other preparations, zinc elicited a bidirectional modulation of GlyRs, with physiological zinc concentrations (1-100 microM) increasing whole cell currents and concentrations >100 microM depressing them. Furthermore, the same concentration of zinc that potentiates I(gly) suppressed currents mediated by the N-methyl-D-aspartate subtype of the glutamate receptor. Thus we provide a pharmacological characterization of native GlyRs expressed by both major neuron types in hippocampus and show that these receptors can be activated by taurine, an amino acid that is highly concentrated in hippocampus. Furthermore, our data suggest that at least two GlyR subtypes are present in hippocampus and that GlyR-mediated currents can be potentiated by zinc at concentrations that suppress glutamate-mediated excitability. PMID- 11877524 TI - Electrical coupling between model midbrain dopamine neurons: effects on firing pattern and synchrony. AB - The role of gap junctions between midbrain dopamine (DA) neurons in mechanisms of firing pattern generation and synchronization has not been well characterized experimentally. We modified a multi-compartment model of DA neuron by adding a spike-generating mechanism and electrically coupling the dendrites of two such neurons through gap junctions. The burst-generating mechanism in the model neuron results from the interaction of a N-methyl-D-aspartate (NMDA)-induced current and the sodium pump. The firing patterns exhibited by the two model neurons included low frequency (2-7 Hz) spiking, high-frequency (13-20 Hz) spiking, irregular spiking, regular bursting, irregular bursting, and leader/follower bursting, depending on the parameter values used for the permeability for NMDA-induced current and the conductance for electrical coupling. All of these firing patterns have been observed in physiological neurons, but a systematic dependence of the firing pattern on the covariation of these two parameters has not been established experimentally. Our simulations indicate that electrical coupling facilitates NMDA-induced burst firing via two mechanisms. The first can be observed in a pair of identical cells. At low frequencies (low NMDA), as coupling strength was increased, only a transition from asynchronous to synchronous single spike firing was observed. At high frequencies (high NMDA), increasing the strength of the electrical coupling in an identical pair resulted in a transition from high-frequency single-spike firing to burst firing, and further increases led to synchronous high-frequency spiking. Weak electrical coupling destabilizes the synchronous solution of the fast spiking subsystems, and in the presence of a slowly varying sodium concentration, the desynchronized spiking solution leads to bursts that are approximately in phase with spikes that are not in phase. Thus this transitional mechanism depends critically on action potential dynamics. The second mechanism for the induction of burst firing requires a heterogeneous pair that is, respectively, too depolarized and too hyperpolarized to burst. The net effect of the coupling is to bias at least one cell into an endogenously burst firing regime. In this case, action potential dynamics are not critical to the transitional mechanism. If electrical coupling is indeed more prominent in vivo due to basal level of modulation of gap junctions in vivo, these results may indicate why NMDA-induced burst firing is easier to observe in vivo as compared in vitro. PMID- 11877525 TI - Spatiotemporal activation of lumbosacral motoneurons in the locomotor step cycle. AB - The aim of this study was to produce a dynamic model of the spatiotemporal activation of ensembles of alpha motoneurons (MNs) in the cat lumbosacral spinal cord during the locomotor step cycle. The coordinates of MNs of 27 hindlimb muscles of the cat were digitized from transverse sections of spinal cord spanning the entire lumbosacral enlargement from the caudal part of L(4) to the rostral part of S(1) segments. Outlines of the spinal cord gray matter were also digitized. Models of the spinal cord were generated from these digitized data and displayed on a computer screen as three-dimensional (3-D) images. We compiled a chart of electromyographic (EMG) profiles of the same 27 muscles during the cat step cycle from previous studies and used these to modulate the number of active MNs in the 3-D images. The step cycle was divided into 100 equal intervals corresponding to about 7 ms each for gait of moderate speed. For each of these 100 intervals, the level of EMG of each muscle was used to scale the number of dots displayed randomly within the volume of the corresponding MN pool in the digital model. One hundred images of the spinal cord were thereby generated, and these could be played in sequence as a continuous-loop movie representing rhythmical stepping. A rostrocaudal oscillation of activity in hindlimb MN pools emerged. This was confirmed by computing the locus of the center of activation of the MNs in the 100 consecutive frames of the movie. The caudal third of the lumbosacral enlargement showed intense MN activity during the stance phase of locomotion. During the swing phase, the focus of activation shifted abruptly to the rostral part of the enlargement. At the stance-swing transition, a transient focus of activity formed in the most caudal part of the lumbosacral enlargement. This was associated with activation of gracilis, posterior biceps, posterior semimembranosus, and semitendinosus muscles. These muscles move the foot back and up to clear the ground during locomotion, a role that could be described as retraction. The spatiotemporal distribution of neuronal activity in the spinal cord during normal locomotion with descending control and sensory inputs intact has not been visualized before. The model can be used in the future to characterize spatiotemporal activity of spinal MNs in the absence of descending and sensory inputs and to compare these to spatiotemporal patterns in spinal MNs in normal locomotion. PMID- 11877526 TI - Computational studies on acquisition and adaptation of ocular following responses based on cerebellar synaptic plasticity. AB - To investigate how cerebellar synaptic plasticity guides the acquisition and adaptation of ocular following response (OFR), a large-scale network model was developed. The model includes the cerebral medial superior temporal area (MST), Purkinje cells (P cells) of the ventral paraflocculus, the accessory optic and climbing fiber systems, the brain stem oculomotor network, and the oculomotor plant. The model reconstructed temporal profiles of both firing patterns of MST neurons and P cells and eye movements. Model MST neurons (n = 1,080) were set to be driven by retinal error and exhibited 12 preferred directions, 30 preferred velocities, and 3 firing waveforms. Correspondingly, each model P cell contained 1,080 excitatory synapses from granule cell axons (GCA) and 1,080 inhibitory synapses. P cells (n = 40) were classified into four groups by their laterality (hemisphere) and by preferred directions of their climbing fiber inputs (CF) (contralateral or upward). The brain stem neural circuit and the oculomotor plant were modeled on the work of Yamamoto et al. The initial synaptic weights on the P cells were set randomly. At the beginning, P cell simple spikes were not well modulated by visual motion, and the eye was moved only slightly by the accessory optic system. The synaptic weights were updated according to integral differential equation models of physiologically demonstrated synaptic plasticity: long-term depression and long-term potentiation for GCA synapses and rebound potentiation for inhibitory synapses. We assumed that maximum plasticity was induced when GCA inputs preceded CF inputs by 200 ms. After more than 10,000 presentations of ramp-step visual motion, the strengths of both the excitatory and inhibitory synapses were modified. Subsequently, the simple spike responses became well developed, and ordinary OFRs were acquired. The preferred directions of simple spikes became the opposite of those of CFs. Although the model MST neurons were set to possess a wide variety of firing characteristics, the model P cells acquired only downward or ipsilateral preferred directions, high preferred velocities and stereotypical firing waveforms. Therefore the drastic transition of the neural representation from the population codes in the MST to the firing rate codes of simple spikes were learned at the GCA-P cell synapses and inhibitory cells-P cell synapses. Furthermore, the model successfully reproduced the gain- and directional-adaptation of OFR, which was demonstrated by manipulating the velocity and direction of visual motion, respectively. When we assumed that synaptic plasticity could only occur if CF inputs preceded GCA inputs, the ordinary OFR were acquired but neither the gain-adaptation nor the directional adaptation could be reproduced. PMID- 11877527 TI - Phase relationships between segmentally organized oscillators in the leech heartbeat pattern generating network. AB - Motor pattern generating networks that produce segmentally distributed motor outflow are often portrayed as a series of coupled segmental oscillators that produce a regular progression (constant phase differences) in their rhythmic activity. The leech heartbeat central pattern generator is paced by a core timing network, which consists of two coupled segmental oscillators in segmental ganglia 3 and 4. The segmental oscillators comprise paired mutually inhibitory oscillator interneurons and the processes of intersegmental coordinating interneurons. As a first step in understanding the coordination of segmental motor outflow by this pattern generator, we describe the functional synaptic interactions, and activity and phase relationships of the heart interneurons of the timing network, in isolated nerve cord preparations. In the timing network, most (approximately 75%) of the coordinating interneuron action potentials were generated at a primary spike initiation site located in ganglion 4 (G4). A secondary spike initiation site in ganglion 3 (G3) became active in the absence of activity at the primary site. Generally, the secondary site was characterized by a reluctance to burst and a lower spike frequency, when compared with the primary site. Oscillator interneurons in G3 inhibited spike activity at both initiation sites, whereas oscillator interneurons in G4 inhibited spike activity only at the primary initiation site. This asymmetry in the control of spike activity in the coordinating interneurons may account for the observation that the phase of the coordinating interneurons is more tightly linked to the G3 than G4 oscillator interneurons. The cycle period of the timing network and the phase difference between the ipsilateral G3 and G4 oscillator interneurons were regular within individual preparations, but varied among preparations. This variation in phase differences observed across preparations implies that modulated intrinsic membrane and synaptic properties, rather than the pattern of synaptic connections, are instrumental in determining phase within the timing network. PMID- 11877528 TI - Model of intersegmental coordination in the leech heartbeat neuronal network. AB - We have created a computational model of the timing network that paces the heartbeat of the medicinal leech, Hirudo medicinalis. The rhythmic activity of this network originates from two segmental oscillators located in the third and fourth midbody ganglia. In the intact nerve cord, these segmental oscillators are mutually entrained to the same cycle period. Although experiments have shown that the segmental oscillators are coupled by inhibitory coordinating interneurons, the underlying mechanisms of intersegmental coordination have not yet been elucidated. To help understand this coordination, we have created a simple computational model with two variants: symmetric and asymmetric. In the symmetric model, neurons within each segmental oscillator called oscillator interneurons, inhibit the coordinating interneurons. In contrast, in the asymmetric model only the oscillator interneurons of one segmental oscillator inhibit the coordinating interneurons. In the symmetric model, when two segmental oscillators with different inherent periods are coupled, the faster one leads in phase, and the period of the coupled system is equal to the period of the faster oscillator. This behavior arises because, during each oscillation cycle, the oscillator interneurons of the faster segmental oscillator begin to burst before those of the slower oscillator, thereby terminating spike activity in the coordinating interneurons. Thus there is a brief period of time in each cycle when the oscillator interneurons of the slower segmental oscillator are relieved of inhibition from the coordinating interneurons. This "removal of synaptic inhibition" allows, within certain limits, the slower segmental oscillator to be sped to the period of the faster one. Thus the symmetric model demonstrates a plausible biophysical mechanism by which one segmental oscillator can entrain the other. In general the asymmetric model, in which only one segmental oscillator has the ability to inhibit the coordinating interneurons, behaves similarly, except only one segmental oscillator can control the period of the system. In addition, we simulated physiological experiments in which a "driving" stimulus, consisting of alternating positive and negative current steps, was used to control a single oscillator interneuron and thereby entrain the activity of the entire timing network. PMID- 11877529 TI - Period differences between segmental oscillators produce intersegmental phase differences in the leech heartbeat timing network. AB - Considerable experimental and theoretical effort has been exerted to understand how constant intersegmental phase relationships are produced between oscillators in segmentally organized pattern generating networks. The phase relationship between the segmental oscillators in the isolated timing network of the leech heartbeat central pattern generator is quite regular within individual preparations. However, it varies considerably among different preparations. Our goal is to determine how the phase relationships in this network are established. Here we assess whether inherent period differences, as suggested by the excitability-gradient hypothesis, play a role in establishing the phase relationships between the two coupled segmental oscillators of the leech heartbeat timing network. To do this we developed methods for reversibly uncoupling the segmental oscillators (sucrose knife) and pharmacological manipulation of the individual oscillators (split bath). Differences in inherent cycle periods between the third and fourth segmental oscillators (G3 and G4) were present in most (20 of 26) preparations. These period differences correlated with the phase differences observed between the segmental oscillators in the recoupled timing network, such that the oscillator with the faster cycle period, regardless of the segment in which it was located, led in phase in proportion to its period difference with the other oscillator. The phase differences between the original (coupled) and recoupled states of individual preparations were similar. Thus application and removal of the sucrose knife did not alter the period difference between the segmental oscillators in the timing network. Pharmacological manipulation of the uncoupled oscillators to alter the period difference between the oscillators led to similar correlated phase differences in the recoupled timing network. Across all experiments the uncoupled segmental oscillator with the faster cycle period established the cycle period of the timing network when recoupled. In conclusion, our findings indicate that an excitability-gradient plays a role in establishing the phase relationship between the segmental oscillators of the leech heartbeat central pattern generator since inherent period differences present between the oscillators are correlated to the phase relationships of the coupled/recoupled timing network. PMID- 11877530 TI - Different mechanisms of calcium entry within different dendritic compartments. AB - From our experiments combining in vivo calcium imaging and electrophysiology on fly vertical motion-sensitive cells (VS-cells) during visual stimulation, we infer different mechanisms of calcium entry within different dendritic compartments; while in the main dendritic branches calcium influx from extracellular space takes place only via voltage-activated calcium channels (VACCs), calcium enters the dendritic tips through VACCs as well as nicotinic acetylcholine receptors (nAChRs). Consequently, neuronal nACHRs of insects have to be assumed to be permeable to some extent for calcium under in vivo conditions. PMID- 11877531 TI - Activity-dependent plasticity of calcium clearance from crayfish motor axons. AB - Previous studies of a crayfish explant culture demonstrated that regenerating motor axons with high impulse activity develop more rapid clearance of cytoplasmic free Ca(2+) than those with low impulse activity. We examined whether Ca(2+) clearance in mature axons also showed activity-dependent plasticity. We studied the phasic and tonic axons of the motor bundle innervating the crayfish closer muscle that display large differences in impulse activity. To compare their Ca(2+) regulation, we applied the Ca(2+) ionophore Br-23187 (1 microM) and measured the increase in intracellular free Ca(2+) concentration ([Ca(2+)](i)) with fura-2. After 55 min of ionophore application, the increase in [Ca(2+)](i) in the phasic axons (1,326 +/- 192 nM) was significantly greater than in the tonic axons (359 +/- 148 nM). This resulted from stronger Ca(2+) clearance in the tonic axon rather than less Ca(2+) influx because blocking Ca(2+) clearance by Na/Ca exchange and mitochondria eliminated these differences in [Ca(2+)](i). Next we determined whether Ca(2+) clearance from the phasic axon could be strengthened by a prolonged increase in impulse activity. The phasic axon was stimulated in vivo at 5 Hz for 1 h/day for 5 days, and 1-3 days after stimulation, Ca(2+) clearance was again examined. After 55 min of Br-23187 (1 microM) exposure, the increase in [Ca(2+)](i) in the stimulated phasic axon was only 232 plus minus 123 nM, which was much less than in the control phasic axons and similar to that in the tonic axons. Thus Ca(2+)-clearance mechanisms adapt to changes in impulse activity both in growing and mature axons. PMID- 11877532 TI - A non-excitatory paradigm of glutamate toxicity. AB - Retinal ganglion cells are driven by glutamatergic synapses, but they are also very susceptible to glutamate toxicity. Whereas the conventional excitotoxicity model of glutamate-induced cell death requires membrane depolarization, we have found that glutamate toxicity need not be linked with excitation. A large subset of ganglion cells possesses high-affinity kainate receptors that are calcium permeable. At 1-5 microM, kainate produced elevation of internal calcium but did not significantly depolarize ganglion cells. This low concentration of kainate caused ganglion cell death, which could be inhibited by specific kainate receptor antagonists. The toxic effect of kainate may be associated with calcium influx, because toxicity was reduced by polyamines that suppress calcium influx and by an inhibitor of calcium phosphatase. Thus activation of ionotropic glutamate receptors can produce neurotoxicity uncoupled from neuroexcitation. PMID- 11877534 TI - Activation of spinobulbar lamina I neurons by static muscle contraction. AB - Spinal lamina I neurons are selectively activated by small-diameter somatic afferents, and they project to brain stem sites that are critical for homeostatic control. Because small-diameter afferent activity evoked by contraction of skeletal muscle reflexly elicits exercise-related cardiorespiratory activation, we tested whether spinobulbar lamina I cells respond to muscle contraction. Spinobulbar lamina I neurons were identified in chloralose-anesthetized cats by antidromic activation from the ipsilateral caudal ventrolateral medulla. Static contractions of the ipsilateral triceps surae muscle were evoked by tibial nerve stimulation using parameters that avoid afferent activation, and arterial blood pressure responses were recorded. Recordings were maintained from 13 of 17 L(7) lamina I spinobulbar neurons during static muscle contraction, and 5 of these neurons were excited. Three were selectively activated only by muscle afferents and did not have a cutaneous receptive field. Spinobulbar lamina I neurons activated by muscle contraction provide an ascending link for the reflex cardiorespiratory adjustments that accompany muscular work. This study provides an important first step in elucidating an ascending afferent pathway for somato autonomic reflexes. PMID- 11877533 TI - Inactivation of semicircular canals causes adaptive increases in otolith-driven tilt responses. AB - Growing experimental and theoretical evidence suggests a functional synergy in the processing of otolith and semicircular canal signals for the generation of the vestibulo-ocular reflexes (VORs). In this study we have further tested this functional interaction by quantifying the adaptive changes in the otolith-ocular system during both rotational and translational movements after surgical inactivation of the semicircular canals. For 0.1-0.5 Hz (stimuli for which there is no recovery of responses from the plugged canals), pitch and roll VOR gains recovered during earth-horizontal (but not earth-vertical) axis rotations. Corresponding changes were also observed in eye movements elicited by translational motion (0.1-5 Hz). Specifically, torsional eye movements increased during lateral motion, whereas vertical eye movements increased during fore-aft motion. The findings indicate that otolith signals can be adapted according to a compromised strategy that leads to improved gaze stabilization during motion. Because canal-plugged animals permanently lose the ability to discriminate gravitoinertial accelerations, adapted animals can use the presence of gravity through otolith-driven tilt responses to assist gaze stabilization during earth horizontal axis rotations. PMID- 11877535 TI - What triggers catch-up saccades during visual tracking? AB - When tracking moving visual stimuli, primates orient their visual axis by combining two kinds of eye movements, smooth pursuit and saccades, that have very different dynamics. Yet, the mechanisms that govern the decision to switch from one type of eye movement to the other are still poorly understood, even though they could bring a significant contribution to the understanding of how the CNS combines different kinds of control strategies to achieve a common motor and sensory goal. In this study, we investigated the oculomotor responses to a large range of different combinations of position error and velocity error during visual tracking of moving stimuli in humans. We found that the oculomotor system uses a prediction of the time at which the eye trajectory will cross the target, defined as the "eye crossing time" (T(XE)). The eye crossing time, which depends on both position error and velocity error, is the criterion used to switch between smooth and saccadic pursuit, i.e., to trigger catch-up saccades. On average, for T(XE) between 40 and 180 ms, no saccade is triggered and target tracking remains purely smooth. Conversely, when T(XE) becomes smaller than 40 ms or larger than 180 ms, a saccade is triggered after a short latency (around 125 ms). PMID- 11877536 TI - A non-alpha7 nicotinic acetylcholine receptor modulates excitatory input to hippocampal CA1 interneurons. AB - The nicotinic acetylcholine receptor (nAChR), particularly the alpha7 subtype, has received profound attention for its role in modifying excitatory postsynaptic currents (EPSCs) in hippocampal pyramidal neurons as well as in neurons from other brain regions. Here, we tested the possibility that an nAChR could affect EPSCs in the interneurons of rat hippocampal slices. Using whole-cell patch-clamp technique on CA1 stratum radiatum interneurons and U-tube application of agents, we show that nicotinic agonists enhance EPSC frequency in interneurons. Among the agents tested, cytisine and mecamylamine were the most effective agonist and antagonist, respectively, suggesting a role for alpha3beta4-containing nAChRs in the modulation of interneuron EPSCs. Ligands selective for the alpha7 nAChR had very little or no effect on interneuron EPSCs. Low concentrations of nicotine also enhanced EPSC frequency, implicating the involvement of non-alpha7 nAChRs in controlling interneuron excitability in smokers. We conclude that nAChR-dependent EPSC modulation in the hippocampus is both subtype- and neuron-specific and that a non-alpha7 nAChR, presumably alpha3beta4, controls glutamate transmission to CA1 interneurons. PMID- 11877537 TI - Network interactions mediated by new excitatory connections between CA1 pyramidal cells in rats with kainate-induced epilepsy. AB - Axon sprouting and synaptic reorganization in the hippocampus are associated with the development of seizures in temporal lobe epilepsy. Synaptic interactions among CA1 pyramidal cells were examined in fragments of hippocampal slices containing only the CA1 area from saline- and kainate-treated rats. Glutamate microapplication to the pyramidal cell layer increased excitatory postsynaptic current (EPSC) frequency, but only in rats with kainate-induced epilepsy. In bicuculline, action potentials evoked in single pyramidal cells increased the frequency of network bursts only in slices from rats with kainate-induced epilepsy. These data further support the hypothesis that excitatory connections between CA1 pyramidal cells increase after kainate-induced status epilepticus. PMID- 11877538 TI - Decoding neural spike trains: calculating the probability that a spike train and an external signal are related. AB - Experimental and clinical applications of extracellular recordings of spiking cell activity frequently are used to relate the activity of a cell to externally measurable signals such as surface potentials, sensory stimuli, or movement measurements. When the external signal is time-varying, correlation methods have traditionally been used to quantify the degree of relation with the neural firing. However, in some circumstances correlation methods can give misleading results. A new algorithm is described that estimates the extent to which a spike train is related to a continuous time-varying signal. The technique calculates the probability of generating a spike train with Poisson statistics if the time varying signal determines the Poisson rate. This is accomplished by successive division of the signal and the spike train into halves and recursive calculation of the probability of each half-signal. The performance of the new algorithm is compared with the performance of correlation methods on simulated data. PMID- 11877539 TI - Modulation of gasp frequency by activation of pre-Botzinger complex in vivo. AB - Under hyperoxic conditions, both chemical stimulation of neurons and focal hypoxia in the pre-Botzinger complex (pre-BotC) in vivo modify the eupneic pattern of inspiratory motor output by eliciting changes in the patterning and timing of phrenic bursts, which includes both phasic and tonic excitation. The influence of this region on the gasping pattern of phrenic motor output produced during severe brain hypoxia is unknown. We therefore examined the effects of chemical stimulation of neurons (DL-homocysteic acid; DLH; 10 mM; < or =20 nl) and focal hypoxia (sodium cyanide; NaCN; 1 mM; < or =20 nl) in the pre-BotC on hypoxia-induced gasping in chloralose-anesthetized, vagotomized, mechanically ventilated cats. Unilateral microinjection of DLH into the pre-BotC during hypoxia-induced gasping increased phrenic burst frequency by approximately 630% (P < 0.01) over baseline frequency due predominantly to a reduction in T(E) (from 28.9 +/- 6.2 to 5.2 +/- 1.8 s; mean +/- SE; P < 0.01). No significant changes in T(I) or rate of rise between hypoxia-induced gasps and the DLH-induced bursts were observed; the effects on peak amplitude of integrated phrenic nerve discharge were variable. Similar responses were evoked by unilateral microinjection of NaCN into the pre-BotC. These findings demonstrate that both activation of pre-BotC neurons and focal hypoxia in the pre-BotC not only influence the eupneic pattern of phrenic motor output but also modify the expression of hypoxia-induced gasping in vivo. These findings also provide additional support to the concept of intrinsic hypoxic chemosensitivity of the pre-BotC. PMID- 11877540 TI - Electronic publishing in medicine: where are we? PMID- 11877541 TI - Pituitary adenylate cyclase activating polypeptide enhances glucose-evoked insulin secretion in the canine pancreas in vivo. AB - OBJECTIVE: To study a local effect of pituitary adenylate cyclase activating polypeptide (PACAP(1-27)) on glucose-evoked insulin release under in vivo conditions. INTERVENTION: Glucose and PACAP(1-27) were locally infused to the pancreas via the superior pancreaticoduodenal artery without interrupting the blood supply. MAIN OUTCOME MEASURES: Plasma insulin and glucose concentrations were determined in samples obtained from the superior pancreaticoduodenal vein and the aorta. Superior pancreaticoduodenal venous blood flow was measured to compute the net output of insulin. RESULTS: PACAP(1-27) (0.005-5 microg) increased the basal insulin secretion by about 15 folds in a dose-dependent manner. Local infusion of either glucose (5%) or PACAP(1-27) (0.05 microg) resulted in a significant increase in the basal insulin output to about 300 microU x min(-1)g(-1), which was highly reproducible upon the second administration of the same dose with an interval of 30 min. When PACAP(1-27) was simultaneously given during glucose infusion, the increased insulin output due to glucose was further enhanced to about 600 microU x min(-1)g(-1). The net increase in PACAP(1-27)-induced insulin output in the presence of glucose was significantly greater than that obtained with PACAP(1-27) alone. There exists a strong and highly significant correlation between changes in glucose level and those in insulin output when both glucose and PACAP(1-27) were administered simultaneously. CONCLUSION: The results indicate that PACAP(1-27) directly enhances the glucose-evoked insulin secretion in the endocrine pancreas in anesthetized dogs. The study suggests that PACAP may play a local facilitating role in insulin secretion in response to glucose loading. PMID- 11877542 TI - Management of gallstone pancreatitis: effects of deviation from clinical guidelines. AB - CONTEXT: Recently published management guidelines for acute pancreatitis provide a standard against which practice can be measured. Specifically it is recommended all patients with gallstone induced pancreatitis have definitive clearance of gallstones within four weeks. OBJECTIVE: To determine if practice in our institution followed these guidelines and to analyse the effects of delayed clearance of gallstones. METHODS: Seventy-six consecutive patients with gallstone pancreatitis presenting within a 15 month period were prospectively studied to compare management with national guidelines and to determine rates of recurrent biliary-pancreatic disease due to delay in clearance of gallstones. RESULTS: Only 5 of 76 patients (6.6%) had operative removal of gallstones within four weeks of their episode of acute pancreatitis. Only 34 of 76 patients (44.7%) had their gallstones removed during the follow up period (minimum 8 months). Fourteen of 76 patients (18.4%) had unplanned readmissions to hospital with biliary-pancreatic disease, necessitating a total of 135 days in hospital. CONCLUSIONS: It is clear from this study that guidelines for the management of gallstone acute pancreatitis are not being met, resulting in high rates of readmission with related disease. PMID- 11877544 TI - Proceedings of the 25th National Congress of the Italian Association of Pancreatology Study (AISP). Cernobbio, Como (Italy). September 20-22, 2001. PMID- 11877543 TI - Estrogen replacement in a protein S deficient patient leads to diarrhea, hyperglucagonemia, and osteonecrosis. AB - CONTEXT: Protein S deficiency and mesenteric venous thrombosis have been described in association with ischemic and/or necrotic bowel. Thrombophilic familial protein S deficiency is known to be amplified by estrogen replacement therapy. Pancreatic ischemia studies have revealed elevated amylase and lipase levels but not hyperglucagonemia. We postulate that estrogen replacement therapy leading to mesenteric and pancreatic ischemia not only caused symptoms of ischemic bowel, but also pancreatic oversecretion of glucagon in a patient with protein S deficiency. Our specific aim was to assess thrombophilic interactions of estrogen replacement therapy and familial protein S deficiency leading to osteonecrosis, hyperglucagonemia, and diarrhea. CASE REPORT: Premarin (2.5 mg/day) was begun following bilateral oophrectomy at age 37. At age 56, hip replacement was done for osteonecrosis of the femoral head. Subsequently, severe epigastric pain and diarrhea developed, which persisted despite conservative measures. Diagnostic evaluation revealed hyperglucagonemia (1420 pg/mL). Although abdominal sonograms, CT scans, and endoscopy failed to document a glucagon secreting tumor, octreotide (50 microg/day) was begun. Normalization of glucagon levels and improvement of abdominal pain was achieved; diarrhea (5-6 episodes/day) persisted. Serologic and genetic testing revealed thrombophilic familial protein S. After stopping estrogen replacement therapy and octreotide, diarrhea and abdominal pain disappeared, glucagon remained normal (normal after 30 months follow-up), and free and functional protein S remained low. CONCLUSIONS: Estrogen induced reduction of protein S, superimposed on familial protein S deficiency, led to osteonecrosis and then, speculatively, to thrombotic mesenteric and pancreatic ischemia with resultant diarrhea, abdominal pain, and hyperglucagonemia. Diarrhea, abdominal pain, and hyperglucagonemia normalized when estrogen was discontinued, and have remained normal over 30 months follow up. PMID- 11877546 TI - The relations among body consciousness, somatic symptom report, and information processing speed in chronic fatigue syndrome. AB - OBJECTIVE: The aim of this study was to assess the potential influence of body consciousness and levels of somatic symptom report upon information processing speed in patients with chronic fatigue syndrome (CFS). BACKGROUND: According to a model of a fixed information processing capacity, it was predicted that in a group of patients with CFS, high body consciousness in combination with a high report of somatic symptoms would affect information-processing speed negatively. METHODS: Information- and motor-processing speed were simultaneously measured with a simple- and a choice-reaction time task, whereas cognitive complaints were rated with two questionnaires. The hypothesized influence of private body consciousness and somatic symptom report upon information-processing speed was tested in a model. A symptom-validity test was used to screen for possible illness behavior. RESULTS: Private body consciousness was directly related to information-processing speed and somatic symptom report. Somatic symptom report was related to both test performance and memory and concentration complaints. CONCLUSIONS: Levels of private body consciousness directly affected somatic symptom report and information-processing speed. This finding supports the role of attentive processes in CFS, and offers, besides possible cerebral dysfunction, an alternative explanation for slowing of information processing in CFS. PMID- 11877547 TI - Plasma hormones and catecholamine metabolites in monozygotic twins discordant for psychosis. AB - OBJECTIVE: To examine plasma cortisol, adrencorticotropin hormone, and indicators of catecholamine activity in monozygotic twins discordant for schizophrenia or major affective disorders. BACKGROUND: Previous research has suggested that catecholamines play a role in the etiology of major mental illness. Several findings have also shown an inverse relation between hippocampal volume and cortisol levels in psychiatric populations including patients who are depressed and patients with Cushing disease. METHOD: In this study, plasma obtained from monozygotic twin pairs discordant for schizophrenia (n = 10) or major affective disorder (n = 3) was assayed for epinephrine, norepinephrine, dihydroxyphenylacetic acid, homovanillic acid, adrencorticotropin hormone, and cortisol. RESULTS: There was significant concordance for levels of dihydroxyphenylacetic acid and homovanillic acid, consistent with the high concordance for indicators of dopamine activity observed in healthy monozygotic pairs. There was also concordance for adrencorticotropin hormone. However, in contrast to findings on healthy monozygotic pairs, there was no relation for epinephrine, norepinephrine, or cortisol. Among patients, there was an inverse correlation between cortisol and the magnitude of the reduction in hippocampal volume, relative to that of the healthy co-twin. CONCLUSION: These findings suggest the potential role of adrenal steroids and hippocampal function in the expression of psychosis. PMID- 11877548 TI - Prediction of clinical response to transcranial magnetic stimulation for depression by baseline lateral visual-field stimulation. AB - OBJECTIVE: We examined whether baseline-affective responses to lateral visual field stimulation could predict clinical responses to left, prefrontal, transcranial, magnetic stimulation (TMS) in patients who are depressed. BACKGROUND: Schiffer et al have reported that left and right lateral visual-field stimulation can often evoke different (positive versus negative) psychologic responses in a given patient. Some had improvements while looking to the left and others while looking to the right. METHODS: We asked 37 patients who were severely depressed and resistant to treatment (26 women, 11 men) to report changes in affective state in response to two pairs of goggles, each allowing vision out of either the left or right visual field. We then evaluated whether these responses predicted clinical responses to TMS as measured by the percent decrease in the Hamilton Depression Rating Scale (HDRS.4%) scores between baseline and 4 weeks (2 weeks after a 2-week course of daily TMS). RESULTS: The 20 patients who felt more improvement with right than left lateral visual-field stimulation with 45-second baseline goggle trials had a 42% (+/- SD 22.2) reduction in HSDR.4%. The 15 patients who felt more improvement looking to the left than to the right had an 11% (+/- SD 28.4) decrease in HSDR.4%. Seventy-five percent of the 20 patients in the first group had a decrease in HSDR.4% more than 20%, and 80% of the 15 in the second group had a decrease in this score of less than 20%. A two-tailed Fisher exact test showed a significant difference between the two groups (p = 0.002). CONCLUSION: Baseline affective responses to lateral visual-field stimulation predicted clinical responses to left, frontal TMS in depressed patients. PMID- 11877549 TI - An adaptation of the modified mini-mental state examination: analysis of demographic influences and normative data: the cache county study. AB - OBJECTIVES: To present a new version of the Modified Mini-Mental State Examination (3MS-R), provide normative information extending to individuals in the 10th decade, and examine the effects of demographic variables on test performance. BACKGROUND: The Modified Mini-Mental State Examination, based originally on the Mini-Mental State Examination, has been used to screen populations for dementia. Providing normative information and an analysis of demographic variables on test performance for this version would support broader use in clinical and other settings. METHODS: Two thousand, nine hundred thirteen elderly individuals determined to be free of dementia and other neurologic and psychiatric conditions served as subjects. An analysis of variance was conducted to examine the effects of age, gender, and education on test performance. Descriptive statistics (means, standard deviations, and percentile ranks) were calculated to summarize the range of normal performance. To examine the sensitivity/specificity of the suggested cut-off points at the 7th and 10th percentiles, two subsamples of elderly individuals, on whom clinical dementia assessments were available, were used to classify individuals with regard to dementia status. RESULTS: Lower age, higher education, and female gender were associated with higher 3MS-R scores. Gender effects were among the weakest, but most important at lower levels of education. Education effects were most prominent in the youngest age groups. Selection of a cut-off point at the 7th percentile revealed 69%-70% sensitivity for detecting dementia, and higher sensitivity for individuals in the youngest age groups. Specificity at this cut off point was 89%. Raising the cut-off point to the 10th percentile improved sensitivity to 73%-76%, but reduced specificity to 85%-86%. CONCLUSION: We present a version of the Modified Mini-Mental State Examination that has demonstrated utility in screening a population for dementia. An analysis of normative information and the effects of demographic influences suggest that the 7th percentile cut-off point performs very well in detecting dementia in 65-79 year-old individuals but less well for individuals in their 80s and 90s. To increase the sensitivity of the 3MS-R to detect dementia or other forms of cognitive impairment, particularly among the "old-old," the test user may wish to raise the cut-off point for impairment in some demographic groups or to supplement the test with additional cognitive measures. PMID- 11877550 TI - Demographic characteristics and normative observations for derived-trail making test indices. AB - OBJECTIVE: The Trail Making Test (TMT) is a psychomotor task that is used routinely in clinical evaluations and consists of two parts: TMT-A and TMT-B. The purpose of the present study was to examine the relations of two derived-TMT indices with age, education, and gender and to establish normative data. BACKGROUND: Prior research has suggested that these derived indices provide purer measures of the executive skills required to complete TMT-B. It has also been suggested that these scores can be effectively used to detect cognitive impairment, and that they are relatively free from the impact of age. METHOD: Difference scores (B minus sign A) and ratio scores (B:A) were computed from the TMT performance of 285 adults who have no history of substance abuse or psychiatric or neurologic disorder and who are not currently being treated with any type of psychotropic medication. RESULTS: Subjects ranged in age from 18 to 90 years and in education level from 6 to 20 years. For the entire sample, (B minus sign A) scores ranged from 6 to 248 seconds (M, 62.15; SD, 49.44), and (B:A) scores ranged from 1.23 to 9.54 (M, 2.81; SD, 1.13). Significant correlations were noted with age and (B minus sign A) and (B:A) scores, accounting for 28% and 6% of the variance, respectively. Subjects were assigned to one of eight age groups, which were formed by decade. Univariate analyses of variance were significant for (B minus sign A) (p < 0.0001) and (B:A) (p < 0.0004) scores. Post hoc analyses indicated that the two oldest age groups had significantly larger (B minus sign A) scores than the younger age groups. However, post hoc analyses with (B:A) scores indicated that only the 70-79-year old age group significantly differed from the three youngest age groups. No other age groups significantly differed from another on this measure. CONCLUSIONS: Normative data are presented for clinical use, and recommendations are made for future research. In addition, the current results suggest that normal aging has a significant impact on cognitive flexibility and indicate that previous suggestions for impairment cut-off scores based on the (B:A) ratio may result in erroneous classification of healthy older adults. PMID- 11877551 TI - Neurocognitive sequelae of exposure to organic solvents and (meth)acrylates among nail-studio technicians. AB - OBJECTIVE: To evaluate neuropsychologic performance among women occupationally exposed to products commonly used in nail studios. BACKGROUND: Organic solvents and (meth)acrylates commonly used in nail studios have known neurotoxic properties. Few studies have examined the potential for cognitive and neurosensory effects of occupational exposure to these substances, and none has addressed exposure occurring in the cosmetics industry. METHODS: Participants in this study included nail-salon technicians (n = 33) and demographically similar controls who had no known history of exposure to toxic chemicals (n = 35). The groups were administered psychologic, neuropsychologic, and neurosensory tests. Aspects of the workplace environment (e.g., square footage of the salon, adequacy of ventilation, and hours worked) also were assessed. RESULTS: Multivariate analysis of variance (MANOVA) revealed that the nail technicians performed more poorly than did controls on tests of attention and processing speed (p = 0.015; eta(2) = 0.20). Olfaction among the nail technicians was below expected performance based on normative data (p < 0.001). A trend toward poorer performance by the nail technicians was observed on the MANOVA investigating executive functioning; individual tests within that domain may be worthy of future investigation (ps = 0.03-0.10). No significant group differences were observed in the domains of learning and memory, visuospatial ability, or fine motor coordination, or on measures of depression and anxiety. Multiple regression indicated that level of occupational exposure as measured by time worked in the industry, adequacy of ventilation, and workplace size predicted 29% of the variance of performance on attentional tasks (p = 0.04). CONCLUSION: Exposure to low-level neurotoxicants common to nail studios may result in mild cognitive and neurosensory changes similar to those observed among solvent-exposed workers in other settings. PMID- 11877552 TI - Cognitive impairments, psychological dysfunction, and coping styles in patients with chronic whiplash syndrome. AB - OBJECTIVE: To investigate underlying mechanisms of cognitive dysfunction in whiplash syndrome, focusing on psychologic factors and coping mechanisms. BACKGROUND: Patients with whiplash often experience cognitive-functioning impairments, although neurologic and neuropsychologic test results do not always support this. Psychologic factors are also hypothesized to influence the existence of whiplash-related cognitive impairments. This has led to a controversy about the etiology of these experiences. METHODS: In this retrospective study, 31 patients with whiplash were compared with 30 neurologic patients and a non-neurologic control group (i.e., subjects with "nonorganic" psychologic symptoms) on memory, attention, and cognitive flexibility. In addition, psychologic questionnaires (Minnesota Multiphasic Personality Inventory, Symptom Checklist, and Utrecht Coping List) were used to investigate psychologic functioning and coping in these patient groups. RESULTS: Patients with whiplash performed similarly to neurologic patients on the cognitive tasks and performed worse on memory and attention tasks compared with the control group. Furthermore, patients with whiplash had high scores on subscales measuring somatization and displayed a predominantly active and palliative coping style. CONCLUSIONS: Somatization, in combination with inadequate coping, might play a role in the development, persistence, or aggravation of whiplash-related symptoms, such as pain or cognitive dysfunction. PMID- 11877553 TI - Musical hallucinations and deafness: a case report and review of the literature. AB - OBJECTIVES: To present the case of a patient with acquired deafness who experienced musical hallucinations (MH) and to conduct a review of the relevant literature. BACKGROUND: Although MH have been known to occur in some people with deafness, literature on this phenomenon, especially from the perspective of etiology and treatment, is limited. METHODS: The case report was prepared using a detailed history, general psychiatric and neurologic examinations, and neurobehavioral assessments, and the pertinent literature from 1965 to 2000 was reviewed. RESULTS: The patient whose case is reported here had acquired deafness and was assessed to have MH and obsessive-compulsive symptoms that responded to the use of nonantipsychotic medication. CONCLUSION: The phenomenon of MH with hearing problems that are either prelingual or acquired can exist separately or be a part of constellation of psychiatric symptoms. No precise etiologic basis is yet clearly identifiable; however, this report offers support for a central mechanism explained by the "Release Theory." Additionally, it appears that such hallucinations might be managed safely with psychotropic medications other than antipsychotics. PMID- 11877555 TI - The modern concept of homeostasis. PMID- 11877557 TI - ["Small volume resuscitation" in hypovolemic rats. Effects on microcirculation]. AB - BACKGROUND: Numerous publications have analysed the hemodynamic effects of "small volume resuscitation" during the initial phases of hemorrhagic shock. Nevertheless nowadays the information about microcirculatory effects are poor. The aim of this study was to estimate the change of tissue perfusion in hypovolemic rats, before and after infusion of Ringer's lactate (RL), hypertonic saline solution (HS) or blood. METHODS: Mesocecal microcirculation was visualized by intravital microscopy during 30 minutes of hemorrhagic hypovolemia (MAP, mean arterial pressure of 40 mmHg) and subsequent reinfusion period. Rats were resuscitated with RL (shed volume), HS (one-seventh of the shed volume), or blood (shed volume). The perfusion was estimated through speed of red blood cells. Moreover MAP, pH and B.E. was measured. RESULTS: Thirty minutes after hemorrhage a very important decrease of capillary flow was noticed and in lesser quantity, of the flow in arterioles and venules. The RL infusion did not cause measureable changes of microcirculatory blood flows. The HS infusion caused an improvement in the flow of arterioles and venules but not in capillaries. The blood infusion caused a progressive improvement in the flow of arterioles, venules and capillaries, however at slightly lower values than previous hemorrhage. CONCLUSIONS: Neither RL nor HS seem as efficient as blood to restore the microcirculatory blood flow in the mesocecum of the rats submitted to hemorrhagic hypovolemia. PMID- 11877556 TI - Antioxidant property of Propofol in the ischemic and reperfused human skeletal muscle. PMID- 11877558 TI - [Impact of an integrated treatment approach of the severely injured patients (ISS =/> 16) on hospital mortality and quality of care]. AB - BACKGROUND: Outcome of severely injured patients is sharply influenced by the level of prehospital and hospital organization. OBJECTIVE: To evaluate the impact of the re-organization of the trauma care process on the quality of care and final outcome of major trauma (ISS =/< 16) victims. SETTING: the Emergency Department (ED) of a 1600 bedded tertiary care hospital. INTERVENTION: a standardized approach to major trauma patients (MT) was implemented: Written protocols were established and trauma teams were organized. All anesthesiologists and trauma surgeons involved in trauma care were enrolled in an educational program including ATLS Courses and the Italian Resuscitation Council Prehospital Trauma Care Course. One of the targets was to assure the early orthopedic stabilization of limb and pelvis fractures. METHODS: Data of all major trauma victims admitted to the ED during 3 comparable periods of time: before (Jan-May 1998), during (Jan-May 1999) and after (Jan-May 2000) the implementation of the process, were retrospectively and prospectively collected and analyzed. RESULTS: MT patients admitted to the hospital increased from 39 in 1998 to 106 in 2000. For similar ISS (30.2 +/- 11.3 in 1998, 29.6 +/- 13.7 in 1999 and 30.5 +/- 12.9 in 2000) hospital mortality dropped from 42% in 1998 to 20.8%. The mean time from hospital admission to surgical orthopedic stabilization was 12 days in 1998, 4.6 in 1999 and 1.3 in 2000. In 2000, 86% of the patients with limbs fractures who required surgical stabilization, were treated within 36 hours from admission vs 11% in 1998. CONCLUSIONS: The implementation of written protocols for trauma care, the organization of trauma teams, educational programs including ATLS and PTC-IRC Courses and a strategy of early stabilization of limb fractures are associated with a dramatic decrease in hospital mortality for major trauma. PMID- 11877559 TI - BIS and Ramsay score in critically ill patient: what future? AB - BACKGROUND: The recent introduction of BIS has improved the concept of utilizing it as a method for the assessment of the sedation level, thus opening new frontiers in critically ill patients care. Its validity in measuring the hypnotic drug effects has been repeatedly proved, but it has rarely been used in ICU. The aim of this paper is to evaluate the correlation between BIS and Ramsay score and its fluctuations with the sedative dosage variations, as a possible utilization in ICU. METHODS: In this study, 20 patients with severe lung disease requiring ventilator support, were recruited. All patients were sedated with propofol and/or midazolam to maintain a Ramsay score of 4. Continuous infusion techniques were commonly used. At time 0 and every half an hour, the patients were evaluated by 2 investigators, using the Ramsay score and the BIS simultaneously. The BIS was calculated by a 1-min recording every time that the sedation score was recorded, for a total of 960 observations. In addition BIS was continuously recorded to establish a baseline value, when the patient was not stimulated. Ventilator settings, medications and vital signs were also recorded. RESULTS: With the increase of the Ramsay score there was a progressive decrease in the BIS score (Ramsay score=2, BIS=88 +/- 15.1; Ramsay score=6, BIS=52.2 +/- 10.7); between BIS and Propofol dosages and between Ramsay score and systolic pressure. CONCLUSIONS: The data obtained show a good correlation between Ramsay score and BIS. PMID- 11877560 TI - Spectrum of acute lung injury, ARDS, sepsis. PMID- 11877561 TI - [Continual epidural therapy in lumbosciatic syndrome. Personal experience]. AB - BACKGROUND: The purpose of this retrospective study was to evaluate the effects of associating several currently used drugs: local anesthetics, corticosteroids, morphine hydrochloride and clonidine administered using continual epidural in lumbosciatic back pain. The association of an opiate and a solution of local anesthetic through infusion increases the analgesic effect and allows smaller quantities of both drugs to be used. The addition of another drug, like clonidine, may allow the dose of the single drugs to be reduced even further, thus resulting in fewer dose-dependent side effects. We know that at least two of the opiate receptor systems and at least three non-opiates modulate the painful sensory afferences. The cortisones can act as both antiphlogistics and antiedemigens, as well as inhibition prostaglandin. METHODS: All the patients in this study were hospitalised after varying periods of home therapy or in other wards. The case histories of 462 patients were studied on admission using routine X-rays and laboratory tests, and also based on a chart divided into parts: the first showed the marks for EO of a patient with lumbosciatica, and the second contained pain-related data. This was measured both directly, using the Scott Huskisson visual parallel and a language card, and indirectly (possible activities, drug intake, etc.). The scale of the analog used defines slight pain with values of less than 44 mm, moderate pain with values between 45-69 mm, strong pain with values between 70-88 mm, and very strong pain above 88 mm. RESULTS: The following results were obtained from this study: successes 87.9%, partial successes 1.7%, failures 10.3%. Of the 48 failures, 31 were operated within a few days, whereas the remaining 17 cases were not operated because the patients declared that they were satisfied with the improvement obtained or gave other reasons. The analgesic effect was frequently observed. The improvement usually increased after the second block so that some patients did not require a third administration. CONCLUSIONS: The results confirm that continual epidural therapy is the most successful and also the most rapid treatment available for pain of lumbar origin. At present we share Finneson's opinion that the same general indications are still valid: 1) discopathy with negative NMR; 2) discopathy with medical contraindications to surgery; 3) discopathy of lumbar and/or radicular origin with alterations to multiple NMR and no reliable signs of a specific root; 4) for symptomatic purposes while waiting for diagnosis and surgery. PMID- 11877563 TI - Microalbuminuria in the US population: third National Health and Nutrition Examination Survey. AB - Microalbuminuria (MA) is associated with adverse health outcomes in diabetic and hypertensive adults. The prevalence and clinical significance of MA in nondiabetic populations is less clear. The purpose of this study was to generate national estimates of the prevalence of MA in the US population. Untimed urinary albumin concentrations (UACs) and creatinine concentrations were evaluated in a nationally representative sample of 22,244 participants aged 6 years and older. Persons with hematuria and menstruating or pregnant women were excluded from analysis. The percent prevalence of clinical proteinuria (UAC > or = 300 mg/L) was similar for males and females. However, the prevalence of MA (urinary albumin creatinine ratio [ACR], 30 to 299 mg/g) was significantly lower in males (6.1%) compared with females (9.7%). MA prevalence was greater in children than young adults and increased continuously starting at 40 years of age. MA prevalence was greater in non-Hispanic blacks and Mexican Americans aged 40 to 79 years compared with similar-aged non-Hispanic whites. MA prevalence was 28.8% in persons with previously diagnosed diabetes, 16.0% in those with hypertension, and 5.1% in those without diabetes, hypertension, cardiovascular disease, or elevated serum creatinine levels. In adults aged 40+ years, after excluding persons with clinical proteinuria, albuminuria (defined as ACR > or = 30 mg/g) was independently associated with older age, non-Hispanic black and Mexican American ethnicity, diabetes, hypertension, and elevated serum creatinine concentration. MA is common, even among persons without diabetes or hypertension. Age, sex, race/ethnicity, and concomitant disease contribute to the variability of MA prevalence estimates. PMID- 11877564 TI - Relationship of serum antioxidant vitamins to serum creatinine in the US population. AB - Several small clinical studies have reported that serum vitamin A levels were higher but serum vitamin C levels were lower among patients with end-stage renal disease. However, the relationship of antioxidant vitamins to renal function has not been studied in the general population. We examined the relationship of serum antioxidant vitamin levels to serum creatinine levels and risk for hypercreatininemia in a representative sample of 6,629 non-Hispanic whites, 4,411 non-Hispanic blacks, and 4,480 Mexican Americans aged 18 years or older who participated in the Third National Health and Nutrition Examination Survey. Serum antioxidant vitamins were measured by isocratic high-performance liquid chromatography, and serum creatinine levels, by the modified kinetic Jaffe method. Serum vitamin A level was positively and significantly associated with serum creatinine level, whereas serum vitamin C level was inversely and significantly associated with serum creatinine level. A one-SD higher level of serum vitamin A (16.9 microg/dL) was associated with a 2.53-fold (95% confidence interval, 1.96 to 3.27; P < 0.001), 2.07-fold (95% confidence interval, 1.84 to 2.33; P < 0.001), and 2.76-fold (95% confidence interval, 1.74 to 4.37; P < 0.001) greater risk for hypercreatininemia among non-Hispanic whites, non Hispanic blacks, and Mexican Americans, respectively. A one-SD higher serum vitamin C level (0.45 mg/dL) was associated with a 22% (95% confidence interval, 0.06 to 0.35; P = 0.01) and 42% (95% confidence interval, 0.08 to 0.62; P = 0.02) lower risk for hypercreatininemia in non-Hispanic whites and Mexican Americans. Our study provides useful information to support the hypothesis that antioxidant vitamins may have an important role in the pathogenesis of chronic renal failure. PMID- 11877565 TI - Evaluation of E-cadherin/catenin complex in primary and secondary glomerulonephritis. AB - Catenins (alpha-, beta-, gamma-catenin, p120(ctn)) are cytoplasmic proteins initially identified in a complex with E-cadherin (ECD). The latter belongs to a superfamily of transmembrane glycoproteins important for cell adhesion in normal and disease states. Catenins and p120(ctn), in particular, are substrates for growth factor receptor tyrosine kinases. Cell adhesive mechanisms have an impact on cell migration and proliferation and thus are potentially involved in the pathogenesis of glomerulonephritides (GNs). Using appropriate monoclonal antibodies, we investigated the immunohistochemical expression of ECD, alpha catenin, beta-catenin, gamma-catenin, and p120(ctn) in renal biopsy specimens from 95 patients with primary GN (n = 51) and secondary lupus-associated GN (n = 44). Examined cases were divided into two groups (proliferative [n = 35] and nonproliferative [n = 60] GNs). Among examined molecules, p120(ctn), beta catenin, and gamma-catenin were expressed more frequently in glomerular epithelial cells, mainly in parietal epithelium (76%, 48%, and 40%, respectively). p120(ctn) and gamma-catenin epithelial expression appeared to be linked closely with proliferative lupus-associated GNs (P = 0.050 and P = 0.029, respectively). Mainly in lupus GNs, with regard to cellular crescents and epithelial cells around microadhesions to Bowman's capsule, p120(ctn) (63% and 73%, respectively), beta-catenin (72% and 75%), and gamma-catenin (75% and 64%) showed the greatest frequencies of positive detection. Mesangial cells were positive only occasionally for the examined molecules. In proliferative lupus GNs, expression of beta-catenin in mesangial cells tended to be prominent (P = 0.066). ECD and alpha-catenin were not expressed in cellular crescents or microadhesions, whereas only ECD was barely detectable in glomerular epithelial cells. In conclusion, expression of beta-catenin, gamma-catenin, and p120(ctn) is focused on glomerular epithelium, as well as on such lesions deriving from it as cellular crescents. This expression probably is linked with epithelial cells' responses to various mitogens, such as growth factors. PMID- 11877566 TI - Significance of early phenotypic change of glomerular podocytes detected by Pax2 in primary focal segmental glomerulosclerosis. AB - In primary focal segmental glomerulosclerosis (FSGS), phenotypic alteration of podocytes is important for the development of cellular lesions (CLs), which precede glomerular scar formation. WT1 and Pax2 are transcription factors involved in kidney development and phenotypic regulation of glomerular epithelial cells. However, the role of WT1 and Pax2 in the development of CLs in primary FSGS is unclear. Using immunohistochemistry, the expression of WT1, Pax2, and cytokeratin (CK), an epithelial marker never found in normal podocytes, was examined in 35 biopsy samples of primary FSGS. Segmental lesions were categorized as: (1) classic segmental scar (CS), (2) CL, and (3) monolayer epithelial (ME) lesion. In normal glomeruli, WT1 was strongly positive in podocytes and weakly positive in parietal epithelium of Bowman's capsule. Pax2 was strongly positive in parietal epithelium of Bowman's capsule, but never expressed in podocytes. Expression of WT1, Pax2, and CK was scantly positive in CSs. WT1 expression was decreased in CLs compared with unaffected podocytes, but Pax2 and CK were strongly expressed in CLs and podocytes of morphologically unaffected tufts in cases with CLs. WT1 expression was strong, as well as Pax2 and CK, in ME lesions. Clinically, urinary protein levels were significantly greater, and the interval from clinical onset to biopsy was significantly shorter in patients with CLs. These results suggest that re-expression of Pax2 in podocytes resulting in phenotypic change to a different epithelial form is one of the important changes for the development of CLs and ME lesions. Alteration from WT1 to Pax2 in podocytes may have an important role in the initiation of glomerular injury in primary FSGS. PMID- 11877567 TI - Add-on angiotensin II receptor blockade lowers urinary transforming growth factor beta levels. AB - Progression of renal failure, despite renoprotection with angiotensin-converting enzyme (ACE) inhibitors in patients with proteinuric nephropathies, may be caused by persistent renal production of transforming growth factor-beta1 (TGF-beta1) through the angiotensin II subtype 1 (AT1) receptors. We tested the hypothesis that AT1-receptor blocker therapy added to a background of chronic maximal ACE inhibitor therapy will result in a reduction in urinary TGF-beta1 levels in such patients. Sixteen patients completed a two-period, crossover, randomized, controlled trial, details of which have been previously reported. All patients were administered lisinopril, 40 mg/d, with either losartan, 50 mg/d, or placebo. Blood pressure (BP) was measured using a 24-hour ambulatory BP monitor. Overnight specimens of urine were analyzed for urine TGF-beta1, protein, and creatinine concentrations. Mean age of the study population was 53 +/- 9 (SD) years; body mass index, 38 +/- 5.7 kg/m2; seated BP, 156 +/- 18/88 +/- 12 mm Hg; and urine protein excretion, 3.6 +/- 0.71 g/g of creatinine. Twelve patients had diabetic nephropathy, and the remainder had chronic glomerulonephritis. At baseline, urinary TGF-beta1 levels were significantly increased in the study population compared with healthy controls (13.2 +/- 1.2 versus 1.7 +/- 1.1 ng/g creatinine; P < 0.001). There was a strong correlation between baseline urine protein excretion and urinary TGF-beta1 level (r2 = 0.53; P = 0.001), as well as systolic BP and urinary TGF-beta1 level (r2 = 0.57; P < 0.001). After 4 weeks of add-on losartan therapy, there was a 38% (95% confidence interval [CI], 16% to 55%) decline in urinary TGF-beta1 levels (13.3 [95% CI, 11.4 to 15.5] to 8.2 pg/mg creatinine [95% CI, 6.2 to 10.7]). The reduction in urinary TGF-beta1 levels occurred independent of changes in mean urinary protein excretion or BP. Thus, proteinuric patients with renal failure, despite maximal ACE inhibition, had increased urinary levels of TGF-beta1 that improved over 1 month of add-on therapy with losartan. We speculate that dual blockade with losartan and an ACE inhibitor may provide additional renoprotection by decreasing renal production of TGF-beta1. PMID- 11877568 TI - Regression of IgA nephropathy: a repeat biopsy study. AB - Histological cure of immunoglobulin A (IgA) nephropathy has been reported only rarely in adults. To elucidate the reversibility of established IgA nephropathy, we performed a repeat biopsy study. A second biopsy was performed in 35 patients with IgA nephropathy in whom hematuria, an essential finding of IgA nephropathy, had disappeared (proteinuria also had disappeared in 23 patients) after a treatment protocol involving high doses of methylprednisolone and tonsillectomy. The interval between the first and second biopsy was 18 to 138 months (mean, 77.1 months). Mean serum creatinine level was 1.11 +/- 0.35 (SD) mg/dL (range, 0.6 to 1.9 mg/dL) at the time of the first biopsy and 0.96 +/- 0.24 mg/dL at the time of the second biopsy. Mesangial proliferation was significantly reduced in second biopsy specimens (mesangial proliferation score: first-biopsy specimens, 2.49 +/- 0.74; second-biopsy specimens, 0.91 +/- 0.89; P < 0.001). Acute inflammatory glomerular lesions, such as endocapillary proliferations, glomerular tuft necrosis, and cellular crescents, were present in 32 patients in first-biopsy specimens, whereas these were no longer present in any of the second-biopsy specimens. Although no significant difference in percentage of globally sclerotic glomeruli was observed between the first and second biopsy specimens, the percentage of segmentally sclerotic glomeruli was significantly lower in second biopsy specimens (P < 0.001). Interstitial mononuclear cell infiltration was markedly reduced in second-biopsy specimens (P < 0.001). The area of renal cortex affected by interstitial fibrosis and/or edema was significantly reduced in second-biopsy specimens (first-biopsy specimens, 21.4% +/- 20.3%; second-biopsy specimens, 9.6% +/- 11.7%; P < 0.01). The distribution of IgA mesangial deposits had diminished in most patients, and no IgA deposits were seen in second-biopsy specimens from 8 patients. These findings indicate that mesangial proliferation and interstitial changes in IgA nephropathy are reversible to a considerable extent. A histological cure may be obtainable in a considerable proportion of patients, especially if treatment is initiated at a relatively early stage. PMID- 11877569 TI - Steroid responsiveness and frequency of relapse in adult-onset minimal change nephrotic syndrome. AB - To clarify factors influencing the response to corticosteroids and subsequent relapses, 62 Japanese adult patients with minimal change nephrotic syndrome were analyzed retrospectively. Five patients experienced remission spontaneously. Fifty-three patients entered complete remission, 3 patients entered partial remission, and 1 patient showed no response to corticosteroids. Fifty-three patients with complete remission were divided into two groups: 38 early responders who experienced remission completely within 8 weeks after starting treatment and 15 late responders who experienced remission after 8 weeks. Blood urea nitrogen and serum creatinine levels and proteinuria selectivity index at presentation were significantly worse in late than early responders. Relative interstitial volume determined by the point-counting method was significantly greater in late than early responders. Relative interstitial volume showed significant correlations with blood urea nitrogen, serum creatinine, and proteinuria selectivity index values. Thirty-three patients experienced a relapse; 13 patients experienced multiple relapses. Fifty-three patients with remission were divided into three groups: 16 patients who experienced relapse within 6 months after the initial response (early relapsers), 17 patients who experienced relapse after 6 months (late relapsers), and 20 patients who did not experience relapse (nonrelapsers). Mean age at onset was younger in early relapsers than late or nonrelapsers. Age at onset correlated inversely with relapse rate in 53 patients with remission and correlated positively with timing of the first relapse in 33 relapsers. It thus was suggested that impaired renal function and poor selectivity of proteinuria, which might be related to interstitial edema, were factors influencing a slower response to corticosteroids. Younger patients had a greater incidence of relapse and were prone to experience relapse earlier. PMID- 11877570 TI - Cross-sectional study of health-related quality of life in African Americans with chronic renal insufficiency: the African American Study of Kidney Disease and Hypertension Trial. AB - We measured health-related quality of life (HRQL) by using the Medical Outcomes Study 36-Item Short-Form (SF-36) in a cross-sectional study of 1,094 African American men and women with mild to moderate chronic renal insufficiency (mean glomerular filtration rate, 45.7 mL/min/1.73 m2) caused by hypertension before randomization onto the African American Study of Kidney Disease and Hypertension (AASK) Trial. Scales contributing to physical health and a summary measure, the Physical Component Summary (PCS) score (mean, 43.4 +/- 10.9 [SD]), were significantly lower than scales relating to mental health and the Mental Component Summary (MCS) score (51.3 +/- 10.3). All scales (except Role-Physical) and the PCS and MCS were significantly higher in men (44.3 +/- 10.9 and 51.8 +/- 10.0, respectively) than women (41.9 +/- 10.8 and 50.5 +/- 10.6, respectively). In multivariate analysis, employment status, education level, household income, body mass index, comorbid medical conditions, years of hypertension, number of antihypertensive drugs prescribed, exercise status, and male sex were significant independent predictors of PCS. Fewer factors predicted MCS and included employment status, marital status, current smoking, age, comorbid medical conditions, and male sex. In the entire AASK cohort, mean scores for individual scales, except Mental Health, and the PCS were lower, but the mean MCS score was slightly higher than values for the US general population. Values for individual scales of the SF-36 and the PCS were substantially higher among AASK participants compared with African-American hemodialysis patients. Six of the eight scales were lower in the AASK cohort compared with groups of racially mixed and exclusively African-American hypertensive subjects. We conclude that physical aspects of quality of life are substantially reduced compared with mental components among AASK participants, and a number of demographic and clinical characteristics significantly impact on HRQL. PMID- 11877571 TI - Increased circulating levels of osteoclastogenesis inhibitory factor (osteoprotegerin) in patients with chronic renal failure. AB - Skeletal resistance to parathyroid hormone (PTH) is one of the major abnormalities underlying bone diseases in uremia, the mechanism of which has not yet been fully elucidated. Osteoclastogenesis inhibitory factor (OCIF), or osteoprotegerin, is a natural decoy receptor for osteoclast differentiation factor (ODF), produced by osteoblasts in response to PTH. To elucidate the kinetics and roles of OCIF in chronic renal failure, serum OCIF levels were measured in 46 predialysis patients and 21 dialysis patients by means of enzyme linked immunosorbent assay (ELISA). Serum OCIF levels in predialysis patients increased as renal function declined (OCIF = 1.178 + 0.233 x creatinine; r2 = 0.413; P < 0.0001). Twenty-four-hour creatinine clearance and 1/OCIF in predialysis patients showed a clear positive correlation and a straight line regression (1/OCIF = 0.443 + 0.004 x creatinine clearance; r2 = 0.425; P < 0.0001). In dialysis patients, serum OCIF levels were significantly elevated (5.18 +/- 1.48 ng/mL) to a level that would inhibit 50% osteoclast formation in vitro. These findings suggest that OCIF accumulates in serum of patients with renal dysfunction. Because serum levels of OCIF with the ability to bind ODF in vitro (active OCIF) correlated well with those of OCIF detected by standard ELISA (active OCIF = 0.251 + 0.877 x OCIF; r2 = 0.829; P < 0.0001), OCIF accumulated in serum may be a candidate uremic toxin responsible for the skeletal resistance to PTH seen in chronic renal failure. Further studies with serum parameters and bone histological evaluation are needed to assess this possibility. PMID- 11877572 TI - Impact of years of dialysis therapy on mortality risk and the characteristics of longer term dialysis survivors. AB - With improving survival and a decreasing probability of receiving a transplant, patients with end-stage renal disease (ESRD) are more likely to remain on hemodialysis therapy for more years than in the past. This study evaluates the effect of years on dialysis (vintage) on relative risk (RR) for death with and without adjustment for comorbidities and treatment factors. It also compares characteristics of patients on hemodialysis therapy for 7 years or longer with those on hemodialysis therapy for 1 to 7 years. Data were combined from two special US Renal Data System studies, the Case Mix Adequacy Study and Waves 1, 3, and 4 of the Dialysis Mortality and Morbidity Study. Excluding the first year of dialysis, 12,687 patients were studied during a 2-year follow-up, censoring at transplantation or loss to follow-up. Unadjusted analysis (vintage 1 to < 2 years as referent) showed that the risk for death remained nearly the same until the end of year 7 of dialysis therapy, after which the risk decreased significantly. However, with adjustment for demographics, comorbidities, and treatment factors, vintage was significantly associated with increased mortality risk during years 2 to less than 8 (RR = 1.12 to 1.30; P < 0.05). Vintage was independently associated with increased adjusted mortality among patients with and without diabetes until approximately 6 to less than 8 years of dialysis therapy. Patients on dialysis therapy for 7 years or longer were significantly (P < 0.05) more likely to be women, younger, and have lower phosphorus levels, higher hematocrits, and higher delivered dialysis doses. We conclude that adjusted mortality risk does not decrease with years on dialysis therapy, and modifiable factors deserve greater attention to improve survival among patients with ESRD with and without diabetes treated by hemodialysis. PMID- 11877573 TI - Increased QTc dispersion and mortality in uremic patients with acute myocardial infarction. AB - QT dispersion (the difference between maximum and minimum QT across the 12-lead electrocardiogram [ECG]), which reflects regional variations in ventricular repolarization, is a predictor of arrhythmia and cardiovascular mortality. The present study was undertaken to assess the difference in QT dispersion between uremic and nonuremic patients with acute myocardial infarction (AMI) and its relationship to post-AMI clinical outcome. Twelve-lead ECG recordings were obtained the first and third days after the onset of AMI in 21 uremic and 21 nonuremic patients. QT intervals were measured on 12-lead ECGs and corrected by heart rate (QTc). Our findings show that uremic patients with AMI had greater QTc dispersion (84 +/- 35 versus 55 +/- 15 milliseconds; P < 0.001), a greater 1-year mortality rate (48% versus 18%; P = 0.003), and underwent fewer reperfusion therapies (5 of 21 versus 17 of 21 patients; P = 0.002) compared with nonuremic patients with AMI. Patients with AMI who died had greater QTc dispersion than those who survived (102 +/- 40 versus 67 +/- 40 milliseconds; P = 0.015). An optimal QTc dispersion cutoff value of 60 milliseconds had a sensitivity of 100% and specificity of 55% in predicting 1-year mortality in uremic patients with AMI. Uremic patients with AMI administered thrombolytic therapies (n = 5) had reduced 1-year mortality rates (0% versus 63%; P = 0.003) and shortened QTc dispersion from days 1 to 3 (changes in QTc dispersion between days 1 and 3, 29% +/- 9% decrease versus 13% +/- 5% increase; P = 0.001) compared with those without therapies (n = 16). Our findings suggest that greater QT dispersion is associated with greater total mortality, and thrombolytic therapies could reduce QTc dispersion and mortality in uremic patients with AMI. It is prudent to refine our current management regimen for uremic patients with AMI to improve the poor clinical outcome. PMID- 11877574 TI - Epidemiology of hemodialysis vascular access infections from longitudinal infection surveillance data: predicting the impact of NKF-DOQI clinical practice guidelines for vascular access. AB - National Kidney Foundation-Dialysis Outcomes Quality Initiative (NKF-DOQI) Vascular Access Guidelines 29 (40% of prevalent patients should have a native fistula) and 30 (<10% use of catheters for chronic hemodialysis) are currently based on opinion, rather than published evidence. The impact of these guidelines on reducing vascular access infection rates is unknown and was tested using data from an outpatient prospective cohort analysis. Patients undergoing hemodialysis from January 1998 through December 2000 at six outpatient facilities in Idaho and Oregon were evaluated prospectively for vascular access infections. There were 111,383 dialysis sessions (DSs) with 471 infections identified (4.2 infections/1,000 DSs). The risk for infection relative to arteriovenous (AV) fistulae was highly dependent on type of access used: 2.2 (P = 0.002) for AV grafts, 13.6 (P < 0.0001) for tunneled catheters, and 32.6 (P < 0.0001) for temporary catheters. Based on incidence infection rates, the number of infections predicted to occur with implementation of guidelines 29 and 30 in this population was calculated, and the percentage of reduction in infection was determined. Following either guideline 29 or 30 alone would have predictably prevented 103 or 97 total infections (22% and 21% reduction) and 40 or 51 bloodstream infections (24% and 30% reduction), respectively. Following both guidelines simultaneously would have prevented 151 total infections (32% reduction) and 64 bloodstream infections (38% reduction). These epidemiological data firmly establish that a major risk for vascular access infections is the type of access used (temporary catheters > tunneled catheters > AV grafts > AV fistulae). Furthermore, they strongly support the role of these NKF-DOQI guidelines in preventing infectious complications attributed to vascular access. PMID- 11877575 TI - Urea kinetics during sustained low-efficiency dialysis in critically ill patients requiring renal replacement therapy. AB - Continuous renal replacement therapies have practical and theoretical advantages compared with conventional intermittent hemodialysis in hemodynamically unstable or severely catabolic patients with acute renal failure (ARF). Sustained low efficiency dialysis (SLED) is a hybrid modality introduced July 1998 at the University of Arkansas for Medical Sciences that involves the application of a conventional hemodialysis machine with reduced dialysate and blood flow rates for 12-hour nocturnal treatments. Nine critically ill patients with ARF were studied during a single SLED treatment to determine delivered dialysis dose and the most appropriate model for the description of urea kinetics during treatment. Five patients were men, mean Acute Physiology and Chronic Health Evaluation (APACHE) II score was 28.9 and mean weight was 92.5 kg. Kt/V was determined by the reference method of direct dialysate quantification (DDQ) combined with an equilibrated postdialysis plasma water urea nitrogen (PUN) concentration and four other methods that were either blood or dialysate based, single or double pool, or model independent (whole-body kinetic method). Solute removal indices (SRIs) were determined from net urea removal and urea distribution volume supplied from DDQ (reference method) and by mass balance using variables supplied from blood based formal variable-volume single-pool (VVSP) urea kinetic modeling. Equivalent renal urea clearances (EKRs) were calculated from urea generation rates and time averaged concentrations for PUN based on weekly mass balance with kinetic variables supplied by either DDQ (reference method) or formal blood-based VVSP modeling. Mean Kt/V determined by the reference method was 1.40 and not significantly different when determined by formal VVSP modeling, DDQ using an immediate postdialysis PUN, or the whole-body kinetic method. Correction of single-pool Kt/V by a Daugirdas rate equation did not yield plausible results. Mean SRI and EKR by the reference methods were 0.61 and 24.8 mL/min, respectively, and not significantly different when determined by blood-based methods. A single-pool urea kinetic model adequately described intradialytic PUN profiles, indicating that SLED was associated with minimal urea disequilibrium. This was supported by the parity between hemodialyzer and whole-body urea clearances, and the mean postdialytic urea rebound of 4.1% (P = 0.13 versus zero). Additional prospective studies are needed in this setting to define the optimal method for dialysis quantification, targets for azotemic control, and optimal modality of renal replacement therapy. In conclusion, SLED delivers a high dose of dialysis with minimal associated urea disequilibrium and can be quantified by Kt/V, SRI, and EKR from blood-based methods using single-pool urea kinetic models. PMID- 11877576 TI - Renal contraction therapy for enlarged polycystic kidneys by transcatheter arterial embolization in hemodialysis patients. AB - Kidneys of patients with autosomal dominant polycystic kidney disease (ADPKD) usually continue to increase in size, even after patients begin dialysis therapy, and the mass effects may lead to severe complications. Such external conventional therapies as surgical and laparoscopic procedures have not yielded satisfactory results. Because kidneys in patients with ADPKD usually are supplied by well developed arteries, we attempted renal contraction therapy in patients with ADPKD by renal transcatheter arterial embolization (TAE) using intravascular coils. After obtaining informed consent, we selected anuric patients on dialysis therapy with markedly distended abdomens or macroscopic hematuria. Between October 1996 and December 2000, a total of 64 patients were treated. Renal size, abdominal circumference, dry weight, hematocrit, and insulin-like growth factor-I were measured before TAE and 3, 6, and 12 months after TAE. Renal sizes decreased to 73.8% +/- 12.0%, 61.7% +/- 14.7%, and 53.4% +/- 11.6% of preinterventional values at 3, 6, and 12 months after therapy, respectively (P < 0.0001). Abdominal circumference and dry weight were significantly decreased at 3, 6, and 12 months (P < 0.0001) compared with baseline values before therapy. Hematocrits increased sequentially after 3, 6, and 12 months (P < 0.0001). Levels of insulin-like growth factor-I an index of nutritional status, significantly increased at 3, 6, and 12 months compared with the baseline value (P < 0.001). This therapy was effective for all patients. Serious complications were not seen after this treatment, although such minor complications as fever and flank pain were observed within the first week after the procedure. Our internal treatment with TAE is a safe and effective procedure that has resulted in improvement in the quality of life and nutritional status of patients with ADPKD. PMID- 11877577 TI - Osteoprotegerin and parathyroid hormone as markers of high-turnover osteodystrophy and decreased bone mineralization in hemodialysis patients. AB - Osteoprotegerin (OPG) has a profound inhibitory effect on osteoclast differentiation and bone resorption. Because high-turnover renal osteodystrophy (ROD) is characterized by increased osteoclast activity, serum OPG concentrations might be used to distinguish between forms of ROD. Twenty-six patients on maintenance hemodialysis therapy underwent a transiliac crest biopsy for evaluation of histopathologic characteristics and histomorphometric studies. ROD was diagnosed as type II (normal or low turnover) or type III (high turnover plus osteoidosis) disease. Bone mineralization density distribution (BMDD) was characterized by measuring the mean trabecular calcium concentration in the biopsy specimen with quantitative backscattered electron imaging. Patients underwent additional dual-energy x-ray absorptiometry (DEXA) of the spine and hip and measurement of such biochemical markers of bone turnover as OPG, intact parathyroid hormone (iPTH), osteocalcin, calcitonin, bone alkaline phosphatase, and cross-laps. OPG levels were significantly reduced in patients with ROD III compared with ROD II (118 +/- 38 versus 204 +/- 130 pg/mL; P < 0.05) and correlated with BMDD (r = 0.43; P < 0.05). Patients with ROD III showed significantly lower BMDD compared with healthy controls (21.42% +/- 0.12% versus 22.17% +/- 0.81% weight; P < 0.01). Besides iPTH, which showed significantly greater levels in patients with ROD III than ROD II (382 +/- 322 versus 136 +/- 156 pg/mL; P < 0.05), none of the serological markers or DEXA was useful in separation of the groups. Discriminant function analysis showed that a combination of OPG and iPTH correctly classifies ROD II in 72% and ROD III in 88% of patients. We conclude that OPG in combination with iPTH can be used as a marker for noninvasive diagnosis of ROD in hemodialysis patients. Furthermore, OPG serum levels might be used to estimate trabecular bone mineralization in these subjects. PMID- 11877578 TI - Use of a bovine model to study the role of adhesion molecule CD11/CD18 in hemodialysis-induced neutropenia. AB - The early neutropenia that occurs with cellulose-based dialysis membranes is believed to result from a cascade of immune events: complement activation, engagement of leukocyte adhesion molecules, cytokine release, and leukocyte sequestration. The beta2 integrin CR3 (CD11b/CD18) is upregulated during hemodialysis, binds complement factor iC3b, and mediates leukocyte adhesion to endothelium and leukoaggregation. Despite being invoked in dialysis-induced neutropenia, there is no direct evidence of a role for CD11b/CD18 in the neutropenia. A unique animal model of beta2-integrin deficiency was discovered in calves experiencing recurrent infections and a paucity of leukocytes in infected tissue. We hypothesized that beta2 integrins mediate the neutropenia of dialysis and directly tested this hypothesis using beta2-integrin-deficient calves. Two 3 month old beta2-integrin-deficient and two age-matched Holstein calves were dialyzed using cuprophane dialyzers. Beta2-integrin-deficient calves had less than 2% of normal neutrophil CD18 expression by flow cytometry. Normal calves had a marked decrease in circulating neutrophils (P < 0.05) to 15% of normal 15 minutes into dialysis (total, four treatments), as well as a decrease in monocytes to 39% (P < 0.05) and lymphocytes to 58% (P < 0.05). CD18-deficient calves had an attenuated decrease in neutrophils (65%; P = not significant), monocytes (78%; P = not significant), and lymphocytes (105%; P = not significant) at 15 minutes. These data, although obtained in a small sample, show that a bovine model can be used to study the early neutropenia of dialysis. These data also suggest that a direct role of beta2 integrins may be occurring in this process. PMID- 11877579 TI - Evaluation of intestinal iron absorption by indirect methods in patients on hemodialysis receiving oral iron and recombinant human erythropoietin. AB - Intestinal iron absorption was evaluated in 40 patients on hemodialysis therapy treated over 4 months with 105 mg/d of oral iron and recombinant human erythropoietin (rHuEPO). The effect of iron stores, erythropoietic activity (EA), and route of rHuEPO administration on absorption was evaluated. Iron was administered after basal determinations had been made and was stopped 15 days before obtaining the final determinations. Intestinal iron absorption was calculated by summing the increase in hemoglobin (Hb) iron (iron used for the synthesis of new Hb) and variations in estimated tissue iron reserves (reserves at the end of the study minus basal reserves). Markers of EA included soluble transferrin receptors (sTfRs) and erythron transferrin uptake (ETU). Iron losses caused by dialysis or normal obligatory iron losses were not measured. Hb levels increased from 8.38 +/- 1.4 to 10.75 +/- 1.5 g/dL (P < 0.05). sTfR levels reached their maximum value at 45 days (3.22 +/- 0.84 mg/mL; P < 0.05), and ETU increased from 40 +/- 26 to 61 +/- 39 micromol/L whole blood/d (P = 0.007). Intestinal iron absorption was 238 mg (interquartile range [Q75 to Q25], 255) at 2 months and 348 mg (Q75 to Q25, 475) at 4 months (P =0.02) and correlated positively with hematocrit at the end of the study (r = 0.826; P = 0.0001). No relationship between iron absorption and basal serum ferritin level or EA markers was observed. Intestinal iron absorption was similar regardless of the route of rHuEPO administration. In conclusion, intestinal iron absorption from medicinal iron covers erythropoietic requirements and allows Hb levels to increase significantly. It is proportional to the degree of efficient erythropoiesis reached and independent of tissue iron stores present before treatment, markers of EA, and rHuEPO administration route. PMID- 11877580 TI - Effects of losartan or enalapril on hemoglobin, circulating erythropoietin, and insulin-like growth factor-1 in patients with and without posttransplant erythrocytosis. AB - Both angiotensin-converting enzyme inhibitors and angiotensin II receptor antagonists reduce hemoglobin (Hb) levels in patients with posttransplantation erythrocytosis (PTE). However, their effects in transplant recipients without PTE are not certain, and the mechanism by which they reduce Hb levels in patients with PTE remains unclear. This study evaluated the effects of losartan and enalapril on Hb levels in relation to serum erythropoietin (EPO) and insulin-like growth factor-1 (IGF-1) levels in 8 patients with PTE and 10 patients without PTE. All 18 patients were treated sequentially with 24 weeks of losartan therapy, followed by 24 weeks of enalapril therapy; the two treatment phases were separated by a washout period. Patients with PTE showed significantly greater baseline Hb and IGF-1 concentrations compared with patients without PTE before both losartan and enalapril treatments. Baseline serum EPO levels were similar for patients with and without PTE. Baseline Hb level correlated significantly with IGF-1 level (r = 0.517; P = 0.002), but not with EPO level. Treatment with enalapril, 5 mg, reduced Hb levels more markedly than treatment with losartan, 50 mg, in patients with PTE. In patients without PTE, enalapril, 5 mg, mildly reduced Hb levels, whereas losartan, 50 mg, had no significant Hb-lowering effect. The reduction in Hb levels with enalapril therapy in patients with PTE was associated with a significant reduction in circulating IGF-1 levels, but not EPO levels, whereas losartan reduced Hb levels with no significant change in circulating IGF-1 and EPO levels. In patients without PTE, no significant change was noted in serum EPO and IGF-1 levels with either treatment. The differential Hb-lowering effect with losartan and enalapril treatment in patients with and without PTE suggests that the pathogenesis for PTE is complex and heterogeneous. Different erythropoietic mechanisms may be involved in patients with and without PTE. Further large-scale study is needed to determine the exact interaction between the renin-angiotensin system and regulation of IGF-1 and EPO synthesis and define the exact mechanism by which losartan and enalapril reduce Hb levels. PMID- 11877581 TI - Flawed proposal for universal conscription of cadaveric organs neglects moral, long-term, and societal implications. PMID- 11877582 TI - Conscription of cadaveric organs for transplantation: let's at least talk about it. AB - Renal transplantation is the optimal treatment for many patients with end-stage renal disease, and for people with other end-stage organ diseases, transplantation may offer the only hope for survival. Unfortunately, the ability to deliver this medical miracle is limited by a severe shortage of human organs. As a result, many people with irreversible organ failure die while waiting for an organ to become available. Compounding this tragic situation is the fact that many usable organs are being buried instead of being transplanted because of the relatively low efficiency of cadaveric organ procurement. One of the major barriers to procurement is family refusal. Several proposals designed to increase consent rates have been suggested, but it is highly unlikely that any of them would approach the 100% efficiency of organ procurement that patients with end stage organ failure so desperately need. However, there is a rarely discussed alternative that would likely achieve this important goal---conscription of cadaveric organs. Under this plan, all usable organs would be removed from recently deceased people and made available for transplantation; consent would be neither required nor requested and, with the possible exception of people with religious objections, opting-out would not be possible. In this article, we review the advantages and disadvantages of this approach. We conclude that consent for cadaveric organ removal is not ethically required and that, from an ethical point of view, conscription is actually preferable. PMID- 11877583 TI - Mechanical ventilation and renal function: an area for concern? AB - Mechanical ventilation is a standard component of intensive care unit management of critically ill patients and widely used for respiratory support. Patients requiring ventilation often have renal dysfunction that can occur as a consequence of the underlying disease or be related to the therapy. Although the physiological consequences of mechanical ventilation on pulmonary and cardiovascular function have been extensively studied, its effects on renal function are not as well defined. Previous experimental studies and few clinical reports have shown a significant effect of mechanical ventilation on renal function. This review compiles the information in this area and provides a framework for future investigation in this field. PMID- 11877584 TI - Towards a biological characterization of focal segmental glomerulosclerosis. AB - The primary form of focal segmental glomerulosclerosis (FSGS) has become one of the most common causes of end-stage renal disease in children and adolescents. FSGS was initially considered to be the histological expression of a single disease entity. However, evidence accumulated during the past four decades indicates that FSGS is heterogeneous in nature. It therefore is not surprising that the many therapeutic combinations and permutations that have been tried have yielded variable results in different hands. This has generated substantial confusion and frustration among physicians and patients alike. Recent progress in genetics and molecular biology has opened promising new vistas of investigation. Identification of genes that control components of the glomerular capillary, proteins that form the structural basis of podocytes, and genetic mutations that affect the integrity of these structures has revolutionized our understanding of the glomerular filtration barrier. Substantial progress also has been made in understanding the mechanisms that lead to progression of renal disease and, ultimately, sclerosis. Studies of these factors are likely to yield a mechanistic based classification of FSGS that will allow us to design therapeutic regimens suited to specific subtypes of this disease. PMID- 11877585 TI - Genetics in kidney disease: how much do we want to know? PMID- 11877586 TI - A 57-year-old woman with gross hematuria and bilateral renal masses: an unusual presentation of acute interstitial nephritis. PMID- 11877589 TI - Survival after end-stage renal disease in autosomal dominant polycystic kidney disease. PMID- 11877591 TI - Assessing the diagnostic accuracy of cystatin C as a marker of impaired glomerular filtration rate. PMID- 11877592 TI - Angiocentric immunoproliferative lesions of the lung associated with diffuse renal involvement. AB - A 62-year-old Japanese man presented with high fever, cough, and sputa. Computed tomography (CT) scan of the chest revealed lung infiltrates with air bronchogram of the right middle lobe and mediastinal lymphadenopathy. Bronchoscopic examination was performed, and Mycobacterium avium complex was detected from bronchoalveolar lavage fluid. Although the administration of clarithromycin and levofloxacin improved the patient's symptoms, the lung infiltrates on chest CT scan gradually worsened. Lung biopsy of segments 4 and 8 by video-assisted thorachoscopy revealed angiocentric and angiodestructive massive lymphoplasmocytic infiltrations with multinucleated giant cells, which were compatible with grade II angiocentric immunoproliferative lesions. The patient was found to have deterioration of renal function, and glomerular filtration rate was 32.6 mL/min. His kidneys were enlarged and showed prominent and diffuse uptake of gallium-67 citrate. Percutaneous renal biopsy revealed massive infiltration of CD4+ mononuclear cells, plasma cells, and eosinophils in the interstitium and destruction of normal structure of tubules. Blood vessels were destroyed and replaced by inflammatory cells. The combination chemotherapy achieved a remission, and the patient has remained free of disease at 2 years after onset of the illness. We recommend the imaging of kidneys for diagnosis and following renal biopsy to evaluate the renal involvement of angiocentric immunoproliferative lesions. PMID- 11877593 TI - Herbal remedy-associated acute renal failure secondary to Cape aloes. AB - Use of traditional herbal remedies is common in Africa, and many patients who visit traditional healers do not need to resort to Western medicine. Acute renal failure is one of the most serious complications resulting from the use of traditional remedies, however, which accounts for 35% of all cases of acute renal failure in Africa. Traditional remedies rarely have been analyzed, and little is known about their nephrotoxicity. We report a case of a 47-year-old man from Soweto, South Africa, who developed acute oliguric renal failure and liver dysfunction after ingestion of an herbal remedy. The patient's renal function recovered slowly, and dialysis was discontinued after several weeks, although serum creatinine did not return to the normal range. Mass spectrometric and chromatographic analysis of the herbal remedy used by the patient revealed the presence of Cape aloes, a previously described nephrotoxin. PMID- 11877594 TI - Aristolochic acid-induced Fanconi's syndrome and nephropathy presenting as hypokalemic paralysis. AB - Hypokalemic paralysis rarely is seen as the presenting feature in patients with Fanconi's syndrome. We describe a 60-year-old man who presented with the inability to ambulate on awakening in the morning. The pertinent history revealed he had consumed Chinese herbs for leg edema for 5 months. Physical examination was unremarkable except for extracellular fluid volume depletion and total paralysis of both lower extremities. Laboratory investigation showed hypokalemia (1.8 mEq/L), hyperchloremic metabolic acidosis (Cl-, 111 mEq/L, and HCO3-, 14.0 mEq/L), hypophosphatemia (0.9 mg/dL) with hyperphosphaturia, hypouricemia (1.3 mg/dL) with hyperuricosuria, and glycosuria, consistent with Fanconi's syndrome. Mild renal insufficiency (serum creatinine, 1.7 mg/dL) also was noticed. Blood and urine screens for heavy metals, autoantibodies, and monoclonal gammopathy were negative. A renal biopsy specimen revealed typical findings of aristolochic acid-associated nephropathy. Aristolochic acids were detected in the consumed Chinese herbs. This case highlights that consumption of Chinese herbs containing aristolochic acids may cause Fanconi's syndrome and should be considered as a cause of hypokalemic paralysis. PMID- 11877595 TI - A patient with apolipoprotein E2 variant (Q187E) without lipoprotein glomerulopathy. AB - Four apolipoprotein (apo) E variants have been reported to be associated with lipoprotein glomerulopathy (LPG), which is characterized by type III hyperlipoproteinemia (type III HLP) and proteinuria and frequently leads to nephrotic syndrome. We report the histologic findings in the kidneys of a type III HLP patient with an apo E variant, apo E Toranomon, in which the glutamine at residue 187 in apo E is substituted by glutamic acid (Q187E). The patient also had type 2 diabetes mellitus. No evidence of lipoprotein thrombi suggestive of LPG was detected, however, and the histologic diagnosis was diabetic nephrosclerosis. This unique case illustrates that not all apo E variants result in LPG, and the location of mutations in the apo E protein is one of the important determinants for the development of LPG. PMID- 11877596 TI - Conversion to tacrolimus for the treatment of cyclosporine-associated nephrotoxicity in heart transplant recipients. AB - Many heart transplant recipients experience nephrotoxicity caused by cyclosporine. Tacrolimus has been associated with similar efficacy and safety in heart transplant recipients compared with cyclosporine. It is unknown whether there is any benefit to switching calcineurin inhibition from cyclosporine to tacrolimus in heart transplant recipients with presumed cyclosporine nephrotoxicity. We report five such cases in which this approach was used successfully. In these cases, the heart transplant recipients had bland urine sediments, low urinary sodium concentrations, adequate cardiovascular and systemic hemodynamics, and cyclosporine levels within or below the therapeutic range as defined by heart transplant criteria. The mechanism of renal failure in these patients was believed to be consistent with renal hypoperfusion secondary to cyclosporine-induced renal vasoconstriction. Conversion to tacrolimus resulted in a prompt and significant improvement in serum creatinine concentrations in these patients (P = 0.002). This report shows that conversion to tacrolimus may represent a useful therapeutic strategy to reduce cyclosporine-associated renal failure in recipients of orthotopic heart transplants. PMID- 11877597 TI - Simultaneous malignant hypertension and cardiac tamponade. AB - Malignant hypertension and cardiac tamponade are uncommon but potentially life threatening medical emergencies. Both conditions may be associated with collagen vascular diseases, such as systemic lupus erythematosus. We report a case of acute cardiac tamponade associated with malignant hypertension secondary to lupus nephritis. Immediately after pericardiocentesis, blood pressure declined substantially. Although malignant hypertension is seen with modest frequency in patients with systemic lupus erythematosus, true cardiac tamponade is a less common complication of lupus serositis. Acute, simultaneous presentation of both life-threatening entities and the hemodynamic course have not been described. PMID- 11877599 TI - Lupus membranous glomerulonephritis mimicking hepatitis C-associated nephropathy. AB - About 20% to 40% of hepatitis C virus (HCV)-infected patients have antinuclear antibody (ANA) seropositivity. Despite the high prevalence of ANA seropositivity in HCV infection, only a few cases of systemic lupus erythematosus (SLE) coinciding with HCV infection have been reported. This discrepancy may be the result of underdiagnosing SLE in patients with chronic HCV infection. The possible underdiagnosis of SLE in HCV-infected patients may be of great concern because interferon-based therapy should be avoided in patients with autoimmune diseases. We report a case of a patient with chronic HCV infection who presented with nephrotic syndrome. She had no previous history of SLE. A kidney biopsy specimen revealed characteristic lesions of lupus membranous nephritis. Further serologic studies confirmed the diagnosis of SLE in this patient. Lupus nephropathy should be ruled out in patients with chronic HCV infection who present with kidney disease and moderate-to-high titers of ANA seropositivity because (1) the association between lupus nephritis and HCV infection may not be coincidental and (2) the diagnosis of lupus nephritis has important therapeutic implications in HCV-infected patients. PMID- 11877598 TI - Fanconi's syndrome, acute renal failure, and tonsil ulcerations after colloidal bismuth subcitrate intoxication. AB - A 22-year-old woman ingested 5.4 g of colloidal bismuth subcitrate (CBS) in a suicide attempt. After ingestion, she presented with Fanconi's syndrome and acute renal failure to our unit. On the third day after ingestion, she was anuric. Ulcerations of both tonsils were observed 8 days after intoxication. Sodium-2,3 dimercapto-1-propanesulfonate (DMPS) is shown to be an effective chelating agent of heavy metal intoxications, but it has only a small effect on elimination of bismuth salts in patients with renal insufficiency without hemodialysis. In our case, we initiated hemodialysis and intravenous treatment with DMPS 60 hours after intoxication. By repeated measurements of bismuth concentrations in serum and dialyzed fluid, we showed its successful elimination. Serum bismuth level decreased from 640 microg/L to 15 microg/L within 6 days. With elimination of bismuth, renal function improved, and tonsil ulcerations healed. Hemodialysis was discontinued on day 14. Follow-up examination 6 weeks later showed normal renal function. Clinicians should be aware that acute renal failure and tonsil ulcerations can occur after CBS intoxication. Generally, acute renal failure caused by CBS intoxication is reversible. Treatment with the chelating agent DMPS in combination with hemodialysis is highly effective in reducing the serum bismuth level in patients with acute renal failure. PMID- 11877601 TI - Essential educational content for advanced practice in psychiatric consultation liaison nursing. AB - Psychiatric Consultation Liaison Nursing (PCLN), an advanced practice subspecialty of psychiatric mental health nursing, has evolved over the last 4 decades in response to increased recognition of the importance of psychophysiological interrelationships and their impact on wellness, physical illness, and recovery. PCLN specialists practice in many countries although there are few PCLN programs in schools of nursing. This article defines PCLN, reviews its history, and recommends essential educational content for inclusion in a master's curriculum to prepare graduates for competent and effective PCLN practice. The educational content is based on the work of the PCLN Core Curriculum Task Force of the International Society of Psychiatric Consultation Liaison Nurses and is integrated with the 1990 Standards of Psychiatric Consultation Liaison Nursing Practice and the 2000 Standards of Psychiatric Mental Health Nursing Practice. The relationship of the PCLN educational content to other national curriculum guidelines is discussed in the proposed PCLN curriculum content. PMID- 11877603 TI - Care meanings, expressions, and experiences of those with chronic mental illness. AB - The care meanings, expressions, and experiences of those with a chronic mental illness living in the community were explored with use of Leininger's Theory of Culture Care Diversity and Universality and the Sunrise Model. Results indicate that people with chronic mental illness have identifiable values, norms, and lifeways that set them apart from the dominant culture. Cultural and social structure factors, ethnohistory, and environmental context influence their desired care. Nurses can use this knowledge to provide culturally congruent care in new ways to enhance the quality of life, productivity, and well-being of this subculture. PMID- 11877602 TI - Understanding those who seek frequent psychiatric hospitalizations. AB - In the period after deinstitutionalization, there has been a rise in hospital readmission rates. It is estimated that the readmission rate for individuals who are frequent users of psychiatric inpatient services is approximately 40% to 50% within 1 year of hospital discharge. Attempts to determine predictors of recidivism have identified multiple variables, some of which are mutually contradictory. Furthermore, comparison among studies is difficult given methodological and theoretical limitations. Despite such issues, however, one consistent predictor of frequent rehospitalization is a person's history of past psychiatric hospital admissions. It seems that those who have shown a pattern of seeking inpatient services in the past tend to repeat this treatment-seeking behavior. The aim of this report is to critically examine some of the predictors of rehospitalization. A better understanding of those who engage in the persistent pattern of seeking inpatient services may assist nurses in planning care that is more suited for their needs. PMID- 11877606 TI - Extension block secondary to partial anterior cruciate ligament tear on the femoral attachment of the posterolateral bundle. AB - PURPOSE: The purpose of this investigation was to prove that a partial tear of the anterior cruciate ligament (ACL) at the femoral attachment of the posterolateral bundle can result in mechanical knee locking and trigger the injury mechanism of an isolated ACL injury. TYPE OF STUDY: Case series. METHODS: From February 1993 through June 1999, 19 cases of knee locking with observation of a torn ACL, confirmed under arthroscopy, were investigated. Patients' ages ranged from 22 to 54 years. The causes of injury were sporting activities in 14 patients (8 skiing, 4 soccer, and 2 badminton); slipping and falling in 2 cases; a pedestrian car accident in 2 cases; and an unknown low-velocity injury in 1 case. RESULTS: On physical examination, each patient indicated that the chief complaint was knee pain; all had a locked knee at 5 degree to 20 degree of full extension and 4 cases showed a locked knee at full flexion. Of the 19 cases, Lachman testing was positive in 3. With respect to intra-articular injuries, 3 patients had a tear in the medial meniscus and 1 had a tear in the lateral meniscus. The average time span between arthroscopy examinations was 3 months with a range of 1 to 10 months. Arthroscopic diagnosis and treatment were performed and meniscal tear was ruled out as the cause of locking in all patients. CONCLUSIONS: The mechanisms of injury for isolated ACL were knee hyperextension and internal rotation of lower extremity brought on especially by a low-velocity injury. The partial ACL tears were found on the femoral attachment site of the posterolateral bundle. The torn segment of the ACL was interposed between the lateral femoral condyle and the lateral tibial condyle and acted as a mechanical obstruction, giving rise to the locking symptom. In all 19 cases, the torn portion of the ACL was excised under arthroscopy and the locked knees were treated properly. PMID- 11877604 TI - Contending with "problem behaviors" in the nursing home. AB - Despite the pervasiveness of challenging resident behaviors in U.S. nursing homes, the staff who work in these facilities typically have little training in mental health or behavior management. This article, based on ethnographic research in a U.S. nursing home that served a predominantly psychiatrically disabled clientele, explores how staff conceptualized and dealt with difficult resident behaviors. It investigates formal and informal strategies used by staff to contend with problematic resident behavior, as well as the obstacles--both internal and external--to effective behavior management in this context. PMID- 11877605 TI - Family caregivers and the mental health care system: reality and dreams. AB - Services directed toward supporting the family caregivers of persons with serious and persistent mental illnesses (SPMIs) are virtually nonexistent despite the fact that such interventions may have the potential to improve outcomes for both the caregivers and the clients. This qualitative study expands our understanding of the caregivers' perspective of nonsupportive and supportive aspects of the current mental health care system as well as desired services. The written narrative responses of 76 family caregivers from National Alliance for the Mentally Ill (NAMI) chapters across the United States were analyzed using content analysis. The caregivers focused on relationship issues, service issues, and broader social system issues. PMID- 11877607 TI - Anterior cruciate ligament reconstruction using four-strand semitendinosus and gracilis tendon grafts and metal interference screw fixation. AB - PURPOSE: The purpose of this study was to determine the outcome of 200 anterior cruciate ligament (ACL) reconstructions using hamstring tendons. TYPE OF STUDY: This is a case series reporting on 200 endoscopic procedures for reconstruction of the ACL. METHODS: This study included patients over 18 years old with a healthy controlateral knee, intact posterior cruciate ligament, and without any peripheral surgical procedure or cartilage injury. Patients having undergone prior ligament reconstruction were excluded from the study. The minimum follow-up was 1 year. Clinical review allowed for documentation of International Knee Documentation Committee (IKDC), KT-1000 arthrometer laxity measurement, and isokinetic dynamometric analysis. RESULTS: Quadriceps and hamstring muscle strength loss was less than 17%. At review, overall IKDC evaluation found that 50% of patients graded A, 44% graded B, and 6% were C or D. The differential anterior laxity was graded A for 157 patients with a median of 1 mm. Of the 113 high-performance athletes, 98 (86%) had resumed a preinjury level of sporting activity. CONCLUSIONS: Endoscopic reconstruction using 4-strand hamstring autograft may be considered safe, reliable, and reproducible. Preliminary outcome is fulfilling and this technique corresponds completely to therapeutic fields regarding ACL reconstructions. PMID- 11877608 TI - Juxta-articular bone marrow signal changes on magnetic resonance imaging following arthroscopic meniscectomy. AB - PURPOSE: Postmeniscectomy osteonecrosis of the knee has been reported in the past decade but the etiology remains unclear. Some investigators have indicated that bone marrow signal changes evident on magnetic resonance imaging (MRI) could be early warning signs of osteonecrosis. The purpose of this study was to determine the incidence rate, location, and magnitude of such changes in bone marrow of the knee after arthroscopic meniscectomy, using MRI. TYPE OF STUDY: Cohort analytic study. METHODS: Ninety-three patients with no bone marrow signal abnormalities on preoperative MRI were examined after isolated arthroscopic meniscectomy. There were 51 men and 42 women with an age range of 11 to 62 years (mean, 36.6 years). Of the total, 57 patients underwent partial meniscectomy (34 medial and 23 lateral) and the others total meniscectomy (10 medial and 26 lateral). MRI examinations were performed independently of postoperative knee symptoms, 1 to 24 months after surgery. Bone marrow changes of the treated knees were evaluated by T1- and T2*-weighted MRI. RESULTS: Thirty-two of 93 patients (34%) had bone marrow signal changes in femoral or tibial condyles shown on postoperative MRI. No patients had these changes in the femoral or tibial condyles opposite from the meniscectomy side and, in the majority of cases, the size was less than half that of the condyle. Fifteen of the 44 patients who underwent medial meniscectomy and 17 of the 49 patients who underwent lateral meniscectomy had such changes. The meniscectomy side did not affect the incidence rate, and frequently both femoral and tibial condyles were involved. Age, gender and articular cartilage condition at the surgery were not risk factors. In contrast, the extent of meniscectomy affected the incidence rate. CONCLUSIONS: This study suggests a positive correlation between arthroscopic meniscectomy and postoperative bone marrow signal changes of the knee. PMID- 11877609 TI - Arthroscopic meniscus repair with bioabsorbable arrows. AB - PURPOSE: The clinical results of a single surgeon's experience with Meniscus Arrows (Bionx, Blue Bell, PA) for meniscal repair are reviewed and reported to determine the safety and efficacy of this device. TYPE OF STUDY: Consecutive sample. METHODS: Over a 3-year period, the senior author has used only Meniscus Arrows for all meniscal repairs. All patients who underwent meniscal repair with at least 12 months of follow-up were evaluated for this study. Thirty patients had a meniscal repair, and 29 were available for follow-up. The average age at surgery was 29 years (range, 15 to 45 years) and there were 24 male and 5 female patients; 25 patients had an anterior cruciate ligament (ACL) reconstruction with the meniscal repair, 2 repairs were performed in ACL-deficient knees, and 2 repairs were performed in ACL stable knees. The average follow-up was 24 months (range, 12 to 42 months). RESULTS: The average Lysholm knee scores for ACL reconstruction, ACL-intact, and ACL-deficient knees improved from 47, 6, and 35, respectively, to 91, 96, and 81 postoperatively. Tegner activity scores improved from 2.7, 0, and 1.5, respectively, to 7.4, 6.5, and 4.5 after surgery. There were no surgical complications, no infections, and no neurovascular injuries. Five patients had mild subcutaneous irritation caused by the Arrow tips, but in each case this resolved within 3 to 7 months. There were 2 failures (7%) that required later arthroscopy and partial meniscectomy. One failure was in an ACL deficient knee, and the other was in an ACL-reconstructed knee. CONCLUSIONS: Although the data presented in this report are based on short-term clinical follow-up, the preliminary results of the Meniscal Arrow repair are encouraging. PMID- 11877610 TI - Radial tears associated with cleavage tears of the medial meniscus in athletes. AB - PURPOSE: To evaluate the significance of a small radial tear in the root of the posterior horn of the medial meniscus in an otherwise normal-looking meniscus in individuals who play vigorous sports. TYPE OF STUDY: Retrospective review. METHODS: Arthroscopy was performed in 1,270 patients; 11 patients (0.86%) had a small radial tear in the root of the medial meniscus. Trimming of the tear revealed a large horizontal cleavage tear of the posterior horn and body of the meniscus. The average age of the affected patients was 29.6 years (range, 21 to 45 years), and all were active in sports. Magnetic resonance imaging was of dubious diagnostic value. Three patients had undergone previous arthroscopy at which time the small radial root tear had been noted but was not thought to warrant treatment. RESULTS: All 11 patients returned to their former levels of activity after adequate surgery. CONCLUSIONS: When a radial root tear in the medial meniscus is found in an athletic patient, the edges of the tear should be trimmed, the root of the medial meniscus examined, and any additional torn cartilage resected. PMID- 11877611 TI - Prevention of deep-vein thrombosis in ambulatory arthroscopic knee surgery: A randomized trial of prophylaxis with low--molecular weight heparin. AB - PURPOSE: The risk of deep vein thrombosis (DVT) in patients undergoing arthroscopic knee surgery is not well known. The purpose of this study was to determine the incidence of DVT, to demonstrate the efficacy of a perioperative and postoperative prophylaxis against thromboembolism with use of low--molecular weight heparin (LMWH), and to show the safety and feasibility of LMWH administration. TYPE OF STUDY: Prospective, single-blind, randomized clinical trial. METHODS: There were 218 consecutive outpatients scheduled for ambulatory arthroscopic knee surgery eligible. Of these, 130 patients were randomized to a treatment group with LMWH (dalteparin: 2,500 IU less-than-or-equal70 kg and 5,000 IU >70 kg, started perioperatively and given once daily for 4 weeks; n = 66) and a control group (n = 64) with no prophylaxis. To detect DVT, all patients underwent bilateral compression ultrasonography before and 12 and 31 days after surgery. RESULTS: Among the 130 patients studied, thromboembolism was significantly lower in the treatment than in the control group: 1 of 66 (1.5%) versus 10 of 64 (15.6%); 95% confidence interval, 7.8% to 26.8%; P =.004. Eighty percent of DVT occurred within the first 14 postoperative days. No severe side effects of LMWH were observed. Only 5% of patients refused continued subcutaneous LMWH injections. CONCLUSIONS: In patients undergoing ambulatory arthroscopic knee surgery without antithrombotic prophylaxis, the risk of DVT is high. Perioperative and postoperative prophylaxis with dalteparin is an effective and safe means of reducing this risk. PMID- 11877612 TI - Incomplete bicruciate knee injuries: Results of treatment with isolated anterior cruciate ligament reconstruction. AB - PURPOSE: To determine how the presence of a partial injury to the posterior cruciate ligament (PCL) in conjunction with an anterior cruciate ligament (ACL) tear affects the outcome of ACL reconstruction. TYPE OF STUDY: Case series. METHODS: Between 1990 and 1998, 18 patients with surgically documented complete ACL ruptures and partial PCL tears, referred to as "incomplete bicruciate knee injuries," were treated with arthroscopic ACL reconstruction and PCL observation. Fifteen patients (83%) were available for follow-up evaluation at a mean of 60 months postoperatively. Follow-up evaluation consisted of physical examination, standard radiographs, isokinetic muscle testing, KT instrumented laxity testing, stress radiographs for posterior and total anterior-posterior translation, and Lysholm, Tegner, and IKDC outcome scores. RESULTS: At follow-up, 11 patients had a negative Lachman and 4 had a 1+ Lachman (3 to 5 mm translation). Instrumented arthrometer tests performed at follow-up showed less than 3 mm of increased anterior translation versus the opposite knee on manual maximum testing at 30 degree in 11 patients, and 3 to 5 mm translation in the remaining 4 patients, with an average of 1.3 mm increased translation. Symptomatic posterior instability was present in only 1 patient who had sustained a repeat injury resulting in a complete PCL tear postoperatively. Final IKDC outcome scores were normal or near normal in 13 patients (87%) and abnormal in 2 (13%). The average Lysholm score was 93 at follow-up, and only 1 patient had a score lower than 80. CONCLUSIONS: Overall, the results of treatment of incomplete bicruciate knee injuries in this study compare favorably with the results of isolated ACL reconstruction reported in other series. We conclude that the clinical significance of an arthroscopically defined partial PCL tear in an ACL-deficient knee is minor, and that isolated ACL reconstruction in this setting appears to be effective treatment. PMID- 11877613 TI - A comparison of the healing potential of fibrillated cartilage versus eburnated bone in osteoarthritic knees after high tibial osteotomy: An arthroscopic study with 1-year follow-up. AB - PURPOSE: To determine the healing potential of fibrillated cartilage in osteoarthritic (OA) knees when the mechanical condition is corrected. TYPE OF STUDY: Prospective study. METHODS: Subjects were patients treated with high tibial osteotomy alone for medial-compartment OA knees. They were divided into 2 groups on the basis of the Outerbridge classification; group A consisted of 37 grade IV knees (eburnation group), and group B consisted of 36 grade III knees (fibrillation group). Twelve months after surgery, the repair of joint cartilage was evaluated for arthroscopic finding and histology. RESULTS: Arthroscopic examination of the femoral joint surface showed that, in group A, 9 knees (24%) were grade II, 14 (38%) were grade III, and 14 (38%) were grade IV; in group B, 3 knees (9%) were grade II, 30 (91%) were grade III, and none were grade IV. On the other hand, examination of the tibial joint surface showed that, in group A, 5 knees (16%) were grade II, 15 (47%) were grade III, and 12 (37%) were grade IV; in group B, 3 knees (9%) were grade II, 28 (80%) were grade III, and 5 (11%) were grade IV. Thus, the incidence of cartilage repair was significantly higher in group A than in group B on both femoral (P <.01) and tibial (P <.05) joint surface. Histologic findings showed most of the nonrepaired joint surface of fibrillated cartilage appeared to be hyaline cartilage, but repair tissues of eburnated bone were mainly fibrocartilage. CONCLUSIONS: The repair of fibrillated cartilage was not promoted, but its degeneration was prevented by correction of the mechanical status. Our results show that fibrillated cartilage in OA knees has little potential for repair even though the mechanical status is corrected. PMID- 11877614 TI - LactoSorb plates for rotator cuff repair. AB - PURPOSE: We present our preliminary studies using LactoSorb plates (Lorenz/Biomet, Warsaw, IN) for rotator cuff repair in humans. The use and application of this plate are described as an adjunct to prevent hole migration in trough-tunnel repairs of rotator cuff tears. METHODS: Fifty patients with rotator cuff tears documented by magnetic resonance imaging underwent arthroscopic subacromial decompression followed by a mini-open repair of the rotator cuff. Tears were repaired using a trough and tunnel technique, horizontal mattress sutures, and a standard LactoSorb plate. Follow-up was obtained on 45 patients, 26 male and 19 female, with an average age of 61.6 years. The patients were followed-up and evaluated at 5 days, 1 month, 3 months, 6 months, and a minimum of 1 year postoperatively. All surgeries were performed by the same surgeon and followed a similar rehabilitation protocol. RESULTS: There were 89% excellent results, 9% good results, and 2% fair results. There were no poor results. No significant swelling, tenderness, or erythema was noted at the operative site, and there were no infections. Postoperative radiographs at 3, 6, and 12 months did not show any bone erosion. No repeat surgical procedures were necessary. CONCLUSION: The results of our study support the use of LactoSorb bioabsorbable plates in the repair of rotator cuff tears to prevent hole migration and potentially poor results or failures of repairs. The LactoSorb plate appears to maintain its strength during the healing period of the repair and does not elicit any clinically noticeable inflammatory process. PMID- 11877615 TI - The derotation sign for perioperative diagnosis of significant partial-thickness rotator cuff tears. AB - PURPOSE: To describe and evaluate a simple perioperative test (the derotation sign) that differentiates significant (grade 3) partial-thickness and small full thickness rotator cuff tears from insignificant (grades 1 and 2) partial thickness rotator cuff tears and intact rotator cuffs. TYPE OF STUDY: Sensitivity and specificity study. METHODS: A study was conducted of 123 patients who underwent shoulder arthroscopy for chronic symptomatic rotator cuff disease, each of whom was subjected to a derotation test under general anesthesia. Specifically, the glenohumeral joint was rapidly distended with fluid just before arthroscopy. One of 3 possible observations was made: (1) the arm rotated internally only, (2) the arm initially rotated internally and then externally rotated (positive derotation sign), or (3) the arm did not rotate at all. Arthroscopic findings were then correlated with the derotation test. RESULTS: Forty-one shoulders had no arm rotation on derotation testing; all had rotator cuff tears greater than 2 cm in size. Forty-two shoulders with impingement and no rotator cuff tears, as well as 23 shoulders with grade 1 or grade 2 partial thickness rotator cuff tears showed internal rotation only. Seventeen shoulders had a positive derotation sign; all of these had either a grade 3 partial thickness or a small full-thickness (<1 cm) rotator cuff tear. CONCLUSIONS: The derotation test is useful in the perioperative differentiation of functionally intact rotator cuffs from those with significant tears. The derotation sign is specific for the diagnosis of grade 3 partial-thickness and/or small (<1 cm) full thickness rotator cuff tears. PMID- 11877616 TI - A study of diagnostic reliability in inferior shoulder instability: Measurement of inferior humeral displacement using arthroscopy. AB - PURPOSE: The purpose of this study was to define arthroscopic measurement of inferior humeral displacement, which allows an objectively estimated reduction of capsular volume using arthroscopic techniques. TYPE OF STUDY: Prospective controlled study. METHODS: Based on a 3-grade classification, inferior displacement caused by axial traction was measured arthroscopically at 15 degree of forward flexion and 70 degree of abduction and neutral rotation over the humeral head in 143 patients (146 shoulders) who underwent surgery consecutively; 99 shoulders because of recurrent dislocations and 47 because of pain in the shoulder. RESULTS: Grades II and III were considered pathologic; grade I (from 0 to 1 cm) was only considered doubtfully positive or within physiologic values. Testing showed a specificity of 96.20%. PMID- 11877617 TI - Arthroscopy of the proximal interphalangeal and metacarpophalangeal joints in rheumatoid hands. AB - PURPOSE: To describe our experience with arthroscopy of the proximal interphalangeal (PIP) and metacarpophalangeal (MP) joints and to assess the value of this technique. TYPE OF STUDY: Case series. METHODS: Arthroscopy was performed on 27 PIP joints and 16 MP joints of 21 patients with rheumatoid arthritis (mean age, 47.2 years; range, 26 to 62 years). After arthroscopic examination, 24 joints were treated with joint irrigation only and 19 were treated with arthroscopic synovectomy. All procedures were performed on an outpatient basis. The diameter of the arthroscope was 1.5 mm, and mini-forceps and a mini-shaver system with a 2.5-mm cutter were used for biopsy and synovectomy. We developed new portals for the PIP joints on the dorsal-lateral aspect, more lateral than previously reported portals. RESULTS: The articular cartilage and synovial membrane of the PIP and MP joints were well visualized, and arthroscopy revealed cartilage changes and synovial proliferation. Because the PIP joint space was not wide enough to insert the arthroscope into the palmar cavity, the palmar part of the articular surfaces and the volar synovium could not be inspected. Synovial biopsy of the dorsal joint capsule was easily performed under arthroscopic visualization. Synovectomy of the dorsal joint capsule and both the radial and ulnar recesses was also possible using the 2-portal technique with a mini-shaver system. No intraoperative or postoperative complications were encountered. CONCLUSIONS: Arthroscopy of the PIP and MP joints is useful in assessing articular cartilage and synovium and for synovial biopsy. Arthroscopic synovectomy can be performed safely with the 2-portal technique. PMID- 11877618 TI - Endoscopic surgery in chronic achilles tendinopathies: A preliminary report. AB - PURPOSE: The objective of this prospective study was to evaluate preliminary results of an endoscopic-assisted surgical technique for patients suffering from chronic Achilles tendinopathies. TYPE OF STUDY: Case series. METHODS: Endoscopic operations were performed on 7 consecutive patients involved in recreational sports suffering from chronic Achilles tendon (AT) lesions in whom conservative treatment had failed. Diagnoses included 2 patients with pure peritendinitis, 4 with peritendinitis and degenerative tendinosis, and 1 with a chronic partial tear. Patients were preoperatively and postoperatively evaluated at a mean follow up period of 16 months (range, 6 to 27 months) with a 0-100 points rating system. All patients were studied preoperatively with magnetic resonance imaging (MRI) and 6 were re-evaluated with the same procedure after surgery. All surgical interventions were performed on an ambulatory basis and 5 under local anesthesia. The surgical endoscopic technique consisted of peritenon release and debridement in cases with pure peritendinitis. In addition, 2 longitudinal tenotomies were performed in cases with degenerative tendinosis or partial tears. RESULTS: According to the scoring system used, all 7 patients had improved final outcome after surgery from a mean of 39 points preoperatively to 88 points postoperatively. The patient with an AT partial tear achieved the lowest score. The only complications were a minor hematoma and edema that resolved spontaneously. Postoperative MRI in patients with tendinosis failed to show evidence of degenerative areas. CONCLUSIONS: Endoscopic surgery may be a valid alternative to treat Achilles tendinopathies unresponsive to conservative treatment because of potential lower morbidity. PMID- 11877619 TI - Biomechanical comparison of hamstring and patellar tendon graft anterior cruciate ligament reconstruction techniques: The impact of fixation level and fixation method under cyclic loading. AB - PURPOSE: To mechanically test different reconstruction techniques of the anterior cruciate ligament (ACL) under incremental cyclic loading and to evaluate the impact of the level and method of graft fixation on tensile properties of each technique. TYPE OF STUDY: In vitro biomechanical study. METHODS: Four hamstring and 1 patellar tendon reconstruction techniques were performed on 40 young to middle-aged human cadaveric knees (average age, 39 years). An anterior drawer with increasing loads of 20 N increments was applied at 30 degree of knee flexion. Anatomic, direct interference screw fixation was tested in 2 hamstring and in the patellar tendon groups. Nonanatomic (extracortical) graft anchorage was tested in the remaining 2 hamstring groups with indirect graft fixations on both sides and the combination of indirect tibial and direct femoral fixation. Structural properties were determined throughout the cyclic loading test. RESULTS: The more anatomic reconstruction techniques provided significantly higher structural properties and smaller loss of fixation compared with nonanatomic, extracortical fixation, with indirect repair on both fixation sites resulting in the lowest structural properties. The tibial fixation site was the weakest link in all of the anatomic reconstructions. Patellar tendon fixation with attached bone blocks in both bone tunnels significantly improved construct stiffness and decreased graft slippage. CONCLUSIONS: The results of this study suggest that anatomic fixation should be preferred for anchorage of hamstring tendons and linkage materials should be avoided. Direct soft-tissue fixation with interference screws still allows considerable graft slippage, which can be limited by using a bone block or application of a backup or hybrid fixation, especially on the tibial fixation site. PMID- 11877620 TI - Bilateral endoscopic carpal tunnel releases: Simultaneous versus staged operative intervention. AB - PURPOSE: Open and endoscopic carpal tunnel release techniques have achieved excellent results for treatment of carpal tunnel syndrome. Symptoms frequently occur bilaterally but there are no reports of simultaneous operative intervention. The purpose of this study was to evaluate results in patients who underwent staged bilateral endoscopic carpal tunnel releases and in those who underwent simultaneous bilateral releases. TYPE OF STUDY: Retrospective review. METHODS: The efficacy of simultaneous and staged bilateral endoscopic carpal tunnel releases was evaluated using a retrospective chart review. This included patients who underwent these procedures during a 48-month period. Group A (48 patients) underwent staged procedures; group group B (48 patients) underwent simultaneous procedures. Inclusion criteria were a positive history and physical examination, positive electrical studies, and failure of conservative measures. Single-incision endoscopic releases were performed on an outpatient basis. Early rehabilitation with intermittent splinting was utilized. The analysis included complications, satisfaction, return to work, physician visits, physical therapy sessions, days to maximum medical improvement for all, and average percentage permanent partial impairment for Workers' Compensation patients. RESULTS: A decrease in return to work at regular duty was noted in the simultaneous group compared with the staged group for patients who had not filed Workers' Compensation claims (P =.0158). The simultaneous group had fewer physician visits than the staged group (P =.0002). Overall patient satisfaction was equal. CONCLUSIONS: Simultaneous bilateral endoscopic carpal tunnel releases are well tolerated with mild restrictions and a decrease in cost. PMID- 11877621 TI - EndoButton drill bit failure. AB - A case of an EndoButton drill bit failure associated with anterior cruciate ligament reconstruction using semitendinosus gracilis autograft is reported. The distal 10 mm flutes of the drill bit sheared off prior to graft passage. This event was likely related to repeated use of this bit, which is intended for single use only. This case highlights another potential pitfall associated with anterior cruciate ligament reconstruction. PMID- 11877622 TI - An unusual metallic foreign body in the lateral tibiofemoral compartment. AB - A child presented with pain and swelling of the knee caused by a fall on the snow. Medical imaging showed an unusual metallic foreign body in the lateral tibiofemoral compartment. Arthroscopy revealed the nature of the foreign body; a metallic piece of stone had penetrated skin and cartilage and was fastened in the cartilage of the lateral femoral condyle where it caused damage to the cartilage and the anterior horn of the lateral meniscus. A partial meniscectomy was performed. PMID- 11877623 TI - Newly devised anterior portal technique for arthroscopic bankart repair using suture anchors. AB - Many techniques and instrument systems have been developed for performing arthroscopic Bankart repair using a suture anchor. The suture anchor procedure relies on a secure knot, which does not become entangled in the anterior portal. A newly devised anterior portal technique can solve this problem. We have used this technique for arthroscopic Bankart repair using suture anchor. This technique may be useful for any type of suture materials such as SLAP lesion. PMID- 11877624 TI - A cannula for non-slip knots. AB - In this article, we introduce a modified cannula that has 1 process and 2 notches. We believe that this will allow a surgeon to complete tying of a non slip knot without any slippage. PMID- 11877628 TI - Arthroscopic repair of a type II SLAP lesion using a single corkscrew anchor. AB - The use of a double-looped 5-mm Corkscrew anchor (Arthrex, Naples, FL) enables the surgeon to use a single anchor to perform a secure fixation of both the anterior labrum as well as the biceps insertion in a type II SLAP lesion. The technique involves tying 1 knot through the anterior portal and a second knot through the posterior portal. PMID- 11877629 TI - Stress computed tomography for evaluating posterolateral knee laxity. AB - A new method is presented to quantify the extent of posterolateral knee laxity. Stress computed tomography (CT) is performed to determine external rotation of the tibia on both knees. Manual maximum external torque is applied to the tibia from both feet by an examiner while both femurs are stabilized by a tape at approximately 30 degree of knee flexion. CT images are acquired at this position and side-to-side differences of external rotation are determined from the images by standard computer software. The technique is simple and helps to quantify posterolateral knee laxity before and after surgical procedures of posterolateral knee reconstruction. PMID- 11877630 TI - How to overcome severed sutures of the tibial bone peg in anterior cruciate ligament reconstruction. AB - We report a case of severed sutures of the tibial bone peg during anterior cruciate ligament reconstruction. The graft retracted proximally in the bone tunnel. We describe a simple and aesthetically acceptable method to salvage this rare complication by use of a small arthrotomy through the defect in the remaining patellar tendon. PMID- 11877631 TI - Arthroscopic technique for the knee in morbidly obese patients. AB - Patients with morbid obesity present an additional surgical difficulty for routine knee arthroscopy. Accurate portal placement in these patients is of utmost importance in order to achieve adequate access to the surgical pathology. To facilitate portal placement, we changed our standard surgical technique, establishing operative portals under direct arthroscopic visualization. We present our technique in this article as an option in the approach to treating these patients. PMID- 11877632 TI - Chondrocalcinosis in an isolated suprapatellar pouch with recurrent effusion. AB - We report the case of a 45-year-old man with a long history of chondrocalcinosis and recurrent painless suprapatellar swelling. Over the 3 months before his admission, the swelling had become persistent and extended 10 cm proximal to the patella on the anterior aspect of the thigh. Magnetic resonance imaging revealed an enlarged suprapatellar pouch that was completely separated from the knee joint cavity by a suprapatellar membrane. Arthroscopic treatment consisted of complete removal of the membrane, creating a passage to the articular cavity, and debridement of the synovial calcifications. The patient was followed-up postoperatively for 2 years and no recurrence of the swelling was reported during this period of time. Two potential mechanisms might have been responsible for the persistent swelling: (1) chondrocalcinosis led to effusion of a suprapatellar pouch that was originally isolated from the knee joint cavity, or (2) the suprapatellar pouch was separated from the knee joint cavity as a result of an incomplete involuted suprapatellar plica that became adherent during recurrent inflammation due to chondrocalcinosis. PMID- 11877633 TI - Coincidence of recurrent arthritis and Behcet's disease following anterior cruciate ligament reconstruction. AB - Chronic recurrent inflammation with joint effusion following anterior cruciate ligament (ACL) reconstruction is common. We report an unusual case of a patient also suffering from undiagnosed Behcet's disease. The patient had chronic recurrent joint effusions of the knee 7 years after traumatic rupture of the ACL and autologous tendon reconstruction using a hamstring tendon autograft. Arthroscopic treatment of the notch impingement and therapy of the accompanying rheumatic disease relieved the patient of his ailment. On the one hand, this case report shows the necessity of including chronic rheumatoid joint disease in the differential diagnosis of a persistent impairment of joint homeostasis following ACL reconstruction, and on the other hand it shows the coincidence of Behcet's disease and recurrent joint inflammation in a patient following ACL surgery. PMID- 11877635 TI - The history and current status of tissue engineering: The future of pediatric surgery. AB - Progress in the pediatric surgical treatment of numerous tissue deficits has been achieved through the relatively new field of tissue engineering. Tissue engineering has distinct advantages over native tissue or prosthetic substitution including self-repair and growth with the patient, avoiding multiple surgeries. The application of microfabrication has allowed more precision in the control of cell interactions and resulting tissue architecture. PMID- 11877637 TI - Risk factors for splenectomy in children with blunt splenic trauma. AB - BACKGROUND/PURPOSE: Nonoperative management and splenic preservation have become standards of care for management of pediatric blunt splenic trauma. However, review of the Pennsylvania Trauma Outcome Study (PTOS) registry found that 15% of children with blunt splenic injury still underwent splenectomy. The authors sought to determine the factors that predisposed to splenectomy in this population. METHODS: Between 1993 and 1997, 754 children, ages 0 to 16 years, who sustained blunt splenic trauma were entered in the PTOS database. These patients were stratified into groups according to the mode of management: nonoperative, splenorrhaphy, or splenectomy. Logistic regression was performed to determine factors associated with splenectomy. RESULTS: Overall, 15.1% of patients underwent splenectomy, 7.4% underwent splenorrhaphy, and 77.5% were treated nonoperatively. Spleen injury grade, nonspleen abdominal injuries, Glasgow Coma Scale 3 to 8, and age 15 to 16 years were significant determinants of splenectomy by multivariate analysis. Children treated at pediatric trauma centers (PTC) underwent significantly fewer splenectomies. CONCLUSIONS: Injury grade, but not hemodynamic instability, was a significant independent determinant of splenectomy in children with blunt splenic trauma. Children treated at PTC are less likely to undergo splenectomy. Ongoing analysis of the management of blunt pediatric splenic injury and reduction of unnecessary splenectomies are needed to optimize care for injured children. PMID- 11877636 TI - Energy expenditure in ill premature neonates. AB - BACKGROUND/PURPOSE: The energy needs of critically ill premature neonates undergoing surgery remain to be defined. Results of studies in adults would suggest that these neonates should have markedly increased energy expenditures. To test this hypothesis, a recently validated stable isotopic technique was used to measure accurately the resting energy expenditure (REE) of critically ill premature neonates before and after patent ductus arteriosus (PDA) ligation. METHODS: Six ventilated, fully total parenteral nutrition (TPN)-fed, premature neonates (24.5 plus minus 0.5 weeks' gestational age) were studied at day of life 7.5 plus minus 0.7, immediately before and 16 plus minus 3.7 hours after standard PDA ligation. REE was measured with a primed continuous infusion of NaH(13)CO(3), and breath samples were analyzed by isotope ratio mass spectroscopy. Serum CRP and cortisol concentrations also were obtained. Statistical analyses were made by paired sample t tests and linear regression. RESULTS: The resting energy expenditures pre- and post-PDA ligation were 37.2 plus minus 9.6 and 34.8 plus minus 10.1 kcal/kg/d (not significant, P =.61). Only preoperative energy expenditure significantly (P <.01) predicted postoperative energy expenditure (R(2) = 88.0%). Pre- and postoperative determinations of CRP were 2.1 plus minus 1.5 and 7.1 plus minus 4.2 mg/dL (not significant, P =.34), and cortisol levels were 14.1 plus minus 2.3 and 14.9 plus minus 2.1 microgram/dL (not significant, P =.52). CONCLUSIONS: Thus, critically ill premature neonates do not have elevated REE, and, further, there is no increase in REE evident the first day after surgery. This suggests that routine allotments of excess calories are not necessary either pre-or postoperatively in critically ill premature neonates. Given the high interindividual variability in REE, actual measurement is prudent if protracted nutritional support is required. PMID- 11877638 TI - Ultrasound-guided fetal tracheal occlusion. AB - BACKGROUND/PURPOSE: This study was aimed at examining the feasibility of fetal tracheal occlusion guided exclusively by ultrasonography and at establishing the technical principles of this procedure based on current instrumentation and ultrasound technology. METHODS: Time-dated pregnant ewes underwent a small laparotomy and partial uterine exposure. Under real-time ultrasound guidance, a steerable guide wire was fed into the fetal tracheal lumen. An 8F catheter then was fed around the guide wire into the trachea. This was followed by intratracheal placement of a 2F coaxial angiographic catheter, connected to a detachable silicone balloon at its extremity, which then was inflated with saline and delivered locally. Euthanasia was performed at term. RESULTS: Complete tracheal occlusion was achieved in all fetuses (n = 7) intraoperatively. Operating time needed for tracheal occlusion, once access to the amniotic cavity was established, varied widely, but could be as short 1 minute. Tracheal lumen dimensions outgrew balloon diameter in 57.1% of fetuses (4 of 7), resulting in balloon dislodgment in those animals. No balloon rupture was observed. CONCLUSIONS: (1) Accurate access to the fetal trachea and local delivery of an occlusive detachable silicone balloon are consistently feasible exclusively under real-time ultrasound guidance with commercially available instrumentation. (2) Balloon dimensions are determining factors of displacement risk and must be selected carefully before tracheal delivery. Ultrasound-guided fetal tracheal occlusion may be a preferable alternative to surgical intervention for treatment of fetal pulmonary hypoplasia. PMID- 11877639 TI - The natural history of prenatally diagnosed conjoined twins. AB - BACKGROUND/PURPOSE: Accurate prenatal diagnosis of complex anatomic connections and associated anomalies has only been possible recently with the use of ultrasonography, echocardiography, and fetal magnetic resonance imaging (MRI). To assess the impact of improved antenatal diagnosis in the management and outcome of conjoined twins, the authors reviewed their experience with 14 cases. METHODS: A retrospective review of prenatally diagnosed conjoined twins referred to our institution from 1996 to present was conducted. RESULTS: In 14 sets of conjoined twins, there were 10 thoracoomphalopagus, 2 dicephalus tribrachius dipus, 1 ischiopagus, and 1 ischioomphalopagus. The earliest age at diagnosis was 9 weeks' gestation (range, 9 to 29; mean, 20). Prenatal imaging with ultrasonography, echocardiography, and ultrafast fetal MRI accurately defined the shared anatomy in all cases. Associated anomalies included cardiac malformations (11 of 14), congenital diaphragmatic hernia (4 of 14), abdominal wall defects (2 of 14), and imperforate anus (2 of 14). Three sets of twins underwent therapeutic abortion, 1 set of twins died in utero, and 10 were delivered via cesarean section at a mean gestational age of 34 weeks. There were 5 individual survivors in the series after separation (18%). In one case, in which a twin with a normal heart perfused the cotwin with a rudimentary heart, the ex utero intrapartum treatment procedure (EXIT) was utilized because of concern that the normal twin would suffer immediate cardiac decompensation at birth. This EXIT-to-separation strategy allowed prompt control of the airway and circulation before clamping the umbilical cord and optimized control over a potentially emergent situation, leading to survival of the normal cotwin. In 2 sets of twins in which each twin had a normal heart, tissue expanders were inserted before separation. CONCLUSIONS: Advances in prenatal diagnosis allow detailed, accurate evaluations of conjoined twins. Careful prenatal studies may uncover cases in which emergent separation at birth is lifesaving. PMID- 11877640 TI - Laparoscopic appendectomy: An unnecessary and expensive procedure in children? AB - BACKGROUND/PURPOSE: Laparoscopic Appendectomy (LA) is a safe procedure in adults resulting in shorter hospitalization and sooner return to activity. The relative merits of LA and open appendectomy (OA) are evaluated in this first prospective and randomized study in children. METHODS: A total of 129 children with appendicitis were included. Forty-three boys and 45 girls, age 1 to 16 years, were enrolled. Randomization was determined by sealed assignment card. OA utilized a 3- to 4-cm right lower quadrant, muscle-splitting incision. Wounds were closed without drains. Antibiotics, when used, consisted of gentamycin, clindamycin, and ampicillin. LA was performed by experienced surgeons utilizing a 3-trocar technique with reusable instruments. Twenty-one children (24%) were perforated. Patients were discharged as soon as they were taking a diet and afebrile. Statistical comparisons were by Fisher's Exact and Wilcoxon rank-sum tests. RESULTS: There were no differences in postoperative analgesia, resumption of oral intake, length of hospitalization, return to normal activities, or morbidity. Laparoscopic appendectomy was associated with longer operating times and increased cost. CONCLUSIONS: Laparoscopic appendectomy in children is not associated with the same advantages reported in adults. LA is a more expensive alternative and offers no advantages related to pain relief, length of stay, return to normal activities, or morbidity. PMID- 11877641 TI - Prenatal diagnosis of vascular anomalies. AB - BACKGROUND/PURPOSE: Vascular anomalies are diagnosed prenatally with increasing frequency. The authors reviewed a group of children treated at their center who had an abnormal prenatal diagnosis to determine (1) fetal age at which the vascular anomaly was detected, (2) general diagnostic accuracy, and (3) impact on ante- and postnatal care. Their findings are compared with reported cases and series. The authors clarify appropriate terminology and underscore the need for interdisciplinary participation of specialists in the field of vascular anomalies. METHODS: Patients referred during prenatal life and children with a history of abnormal antenatal findings seen at our vascular anomalies center during a 1-year period (September 1999 through August 2000) were included in this study. The fetal age at diagnosis, pre- and postnatal diagnoses, antenatal course, and neonatal outcome were obtained from the parents, through chart reviews, and through telephone interviews with the treating obstetricians. RESULTS: Twenty-nine patients with vascular anomalies were identified: 17 had a correct prenatal diagnosis, and 12 had an incorrect diagnosis, an overall diagnostic accuracy of 59%. Capillary-lymphatic-venous malformations (CLVM) most often were correctly diagnosed (67%), followed by lymphatic malformation (LM, 62%) and hemangioma (59%). In the infants who received correct diagnoses in utero, there were no fetal deaths and there was no neonatal morbidity. Maternal steroids were administered for a fetus with an intrahepatic hemangioma and deteriorating cardiac function, with subsequent stabilization and successful delivery of a healthy neonate. Among infants with incorrect diagnoses, there was 1 postnatal death, 1 case of erroneous gender assignment, 1 case of unnecessary fetal surgical intervention, 1 unnecessary neonatal laparotomy, and 1 delay in diagnosis of a malignancy. Cesarean section was done for 65% of correctly diagnosed cases, (including 2 ex utero intrapartum [Exit] procedures) and for 33% of incorrectly diagnosed cases. Most diagnoses were made during the mid- to late second trimester and third trimester; only 4 cases (14%) were detected before 20 weeks. CONCLUSIONS: In this series, accurate diagnosis optimized antenatal care by providing an opportunity for planning deliveries, for pharmacologic fetal intervention in 1 case, and for appropriate parental counselling. Inaccurate diagnosis was associated with significantly increased morbidity and mortality. Finally, the intrauterine diagnosis of LM should be distinguished from posterior nuchal translucency, an obstetric term applied to fetal lymphatic abnormalities detected in the first and second trimesters that do not manifest as postnatal LM. PMID- 11877642 TI - Leptin: a new growth factor for the small intestine. AB - PURPOSE: This study was designed to evaluate the potential growth factor effects of systemic administration of leptin on mucosal mass and absorptive function in normal rat intestine. METHODS: Twenty male Sprague-Dawley rats underwent placement of a jugular venous catheter connected to a subcutaneous osmotic pump designed to deliver its contents at a constant rate. The rats were divided into 4 groups (n = 5 per group) based on the contents of the osmotic pump: group 1, 0.1% bovine serum albumin; group 2, leptin, 6.25 microgram/kg/d; Group 3, leptin, 18.75 microgram/kg/d; Group 4, leptin, 43.75 microgram/kg/d. After a 14-day infusion, [(14)C] galactose and [(14)C] glycine absorption were determined using a closed, recirculation technique. DNA content was determined from mucosal biopsies. Total RNA was extracted from mucosal samples, reverse transcribed, and amplified via polymerase chain reaction for the following primer pairs: sodium/glucose cotransporter (SGLT-1), fructose transporter (GLUT-5), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH, internal standard). Statistical analysis was performed by analysis of variance and expressed as mean plus minus SEM. RESULTS: Systemic administration of increasing doses of leptin enhanced DNA content when compared with the appropriate control (group 2, 1.06 plus minus 0.04 [P <.05]; group 3, 1.1 plus minus 0.05 [P <.01]; and group 4, 1.07 plus minus 0.06 [P <.05]. Leptin enhanced mucosal absorptive function (galactose: group 2, 2.31 plus minus 0.15 [P <.01]; group 3, 2.71 plus minus 0.06 [P <.01]; group 4, 2.19 plus minus 0.28 [P <.05]; glycine: group 2, 2.34 plus minus 0.31 [P <.05]; group 3, 3.32 plus minus 0.14 [P <.01]; group 4, 3.1 plus minus 0.27 [P <.01]) in the normal intestine when compared with the appropriate control animals. Also, leptin enhanced the gene expression of the carbohydrate transporters when compared with the appropriate control rats. CONCLUSIONS: These data show that systemic leptin administration enhances mucosal mass and absorptive function in normal rat intestine. Thus, leptin appears to be a growth factor for normal small intestine and may play a role in patients who acquire intestinal dysfunction. PMID- 11877643 TI - Cystic adenomatoid malformation volume ratio predicts outcome in prenatally diagnosed cystic adenomatoid malformation of the lung. AB - BACKGROUND/PURPOSE: Cystic adenomatoid malformation of the lung (CAM) diagnosed in utero has a variable natural history that may result in hydrops in up to 40% or regress in up to 15%. No criteria have been available to determine which lesions would grow and develop hydrops versus those whose growth would stabilize or regress. To better understand the natural history of CAM the authors developed a measure of tumor volume normalized for gestation age, the CAM volume ratio, or CVR. The results of an initial retrospective review of CVR at presentation suggested its usefulness as a predictor of outcome in CAM. The authors now report the results of prospective use of the CVR both to track tumor growth and regression during gestation and confirm its predictive value in fetuses with CAM. METHODS: In the retrospective review performed between November 1998 and August 1999, 32 fetuses with CAM were reviewed and divided into those with hydrops and those in whom hydrops never developed. The CVR was determined by measuring 3 dimensions of the CAM using the formula for the volume of an ellipse and dividing by the head circumference to correct for differences in gestational age. Of the 32 fetuses in the retrospective study, the 8 that had hydrops had a significantly higher CVR (3.1 plus minus 1.1) compared with hydropic fetuses (0.74 plus minus 0.48; P <.001). The mean of the nonhydropic fetus's CVR plus 2 standard deviations (0.74 + 0.96 = 1.7) was used as a cutoff in the subsequent prospective study. From September 1, 1999 through March 1, 2001, the authors evaluated prospectively 58 patients with CAM by CVR measurement. These patients were followed up with serial ultrasound scans, and CVR at presentation correlated with the development of hydrops, survival, need for fetal intervention, and the need for ventilatory support or extracorporeal membrane oxygenation (ECMO), and length of hospital stay postnatally. The indication for fetal intervention was the development of hydops. RESULTS: The fetuses with CVR less-than-or-equal1.6 (n = 42) were considered to be at low risk for the development of hydrops, and those with CVR greater than 1.6 (n = 16) were considered at increased risk for developing hydrops. Of the 42 fetuses in the low-risk group, 7 (16.7%) developed hydrops, and all but 1 had a dominant cyst. If CAMs with a dominant cyst are excluded, only 1 of 36 (2.8%) of CAMs with CVR less-than-or-equal 1.6 developed hydrops (P <.001). In fetuses with CVR at presentation more than 1.6, 12 of 16 (75%; P <.005) developed hydrops. Seventeen fetuses underwent fetal treatment (8 CVR less-than-or-equal 1.6; 9 CVR > 1.6): 7 patients required open fetal surgery (survival rate, 2 of 7), 6 patients thoracoamniotic shunting (survival rate, 6 of 6); and 4 patients cyst aspiration (survival rate, 4 of 4). All survivors of fetal intervention required at a least brief period of ventilatory support; none required ECMO. CONCLUSIONS: A CVR of greater than 1.6 at presentation accurately predicts increased risk of hydrops developing in CAM. A CVR of less-than-or equal1.6 at presentation suggests that the risk of hydrops developing in the absence of a dominant cyst is less than 3%. The CVR is a useful sonographic indicator of fetuses at risk for hydrops who require close ultrasound observation and possible fetal intervention. PMID- 11877644 TI - Urologic injuries associated with repair of anorectal malformations in male patients. AB - BACKGROUND/PURPOSE: Serious injuries to the urinary tract may occur during the repair of an anorectal malformation, especially in boys. This review of a large series of patients characterizes factors that may either lead to, or prevent, those injuries. METHODS: A retrospective review of 1,003 boys with anorectal malformations was performed. RESULTS: A total of 129 injuries in 1,003 patients were identified. Five hundred seventy-two of the 1,003 patients (group A) underwent definitive repair at the authors' institution. In this group, there were 19 urologic injuries; 1 bladder perforation, 1 divided ureter, 2 divided vas defera, 1 prostatic injury, 7 seminal vesicles were opened and closed, and in 7 cases, the urethra was opened and closed during the repair. Follow-up ranges from 15 years to 1 month and no late sequelae have been observed. The second group (B) consisted of 431 patients who underwent various operations at other institutions. In this group, 110 urologic injuries in 97 patients were noted. These included neurogenic bladder (n = 27), persistent, recurrent or acquired recto-urethral fistulae (n = 30), posterior urethral diverticulae that required reoperation (n = 23), urethral injuries leading to stenosis or acquired atresia (n = 19), pull through of major urinary structures (n = 2), injured ureter (n = 1), opened seminal vesicle (n = 1), divided vas defera (n = 4), impotence (n = 1), and loss of ejaculation (n = 2). Several significant associations were noted. The most significant was that all 27 patients with neurogenic bladder and all 19 of those in group B with urethral injuries did not undergo a distal colostogram to define the level of the fistula before repair. Posterior urethral diverticulae were seen only in cases of recto-bulbar urethral fistulae repaired via an abdominal perineal approach. CONCLUSIONS: Significant urologic injuries are associated with the repair of anorectal malformations. The risk of injury is increased in those patients who undergo repair without a distal colostogram. PMID- 11877645 TI - Therapeutic cecostomy tubes in infants with imperforate anus and caudal agenesis. AB - BACKGROUND/PURPOSE: Caudal agenesis is indicative of a poor prognosis for achieving normal bowel function in children with high imperforate anus (HIA). Complications with impaction, soiling, megarectosigmoid, and adverse responses to rectal enemas are inevitable and associated with long-term psychosocial dysfunction. In an attempt to avoid these outcomes, the authors began a prospective evaluation in these high-risk patients of skin level cecostomy tubes placed in infancy. METHODS: Between October 1997 and March 2001, 8 infants with variants of caudal agenesis underwent presumptive therapeutic placement of a cecostomy tube in conjunction with colostomy closure (n = 7) and anal transposition (n = 1). Mean age at time of cecostomy placement was 16 months. Efficacy of daily irrigations with individually modified electrolyte solutions has been evaluated over the subsequent 1 to 41 months with a mean follow-up of 22 months. RESULTS: Effective daily colonic evacuation was achieved in 7 infants. Transient episodes of incomplete emptying often associated with soiling were resolved with adjustments in solution contents and volume. No patient has required admission for impaction, and only 1 patient has required rectal enemas. There have been no major complications, and minor problems with cramping, granulation tissue, and site discomfort quickly resolved. One patient with anal anastamotic stricture consistently has not responded to antegrade irrigation and continues to require intermittent rectal enemas and anal dilation. Attempts to wean the irrigations occur as the infants grow older and are able to participate in toilet training; however, in this preschool population, tapering of the frequency has been tolerated only transiently. CONCLUSIONS: Antegrade enemas via a cecostomy device are highly effective for bowel management in infants with HIA and caudal agenesis. They can be placed with minimal morbidity at the time of initial reconstruction or colostomy closure. Consideration of this procedure in all infants with HIA may be indicated. PMID- 11877646 TI - Sigmoid irrigation tube for the management of chronic evacuation disorders. AB - BACKGROUND/PURPOSE: Antegrade colonic irrigation, in which the right colon is accessed via appendicostomy or cecostomy, now is an important adjunct in the management of children with chronic evacuation disorders. However, in most children, the major area of dysfunction is the left rather than the right colon. The authors developed a simple, percutaneous endoscopic, laparoscopically controlled sigmoid irrigation tube placement and evaluated the results in 4 children. METHODS: A rigid sigmoidoscope is advanced into the upper sigmoid and the loop brought in contact with the abdominal wall under laparoscopic control. A small skin incision is made and a needle pushed across the abdominal and sigmoid walls into the lumen of the sigmoidoscope. A guide wire is advanced through the needle into the scope and retrieved. After the scope is removed, a PEG-type catheter is attached to the guide wire and pulled back, securing the sigmoid loop to the abdominal wall. The tube is subsequently converted to a skin-level device by simply adding an external port valve. RESULTS: All 4 patients achieved prompt evacuation in the sitting position. CONCLUSIONS: Sigmoid tube for antegrade irrigation is an appealing alternative to conventional cecal access. The procedure is simple and may offer physiologic advantages. PMID- 11877647 TI - Differences in outcome with subspecialty care: pyloromyotomy in North Carolina. AB - BACKGROUND/PURPOSE: Proponents of subspecialization in surgery claim that fellowship training improves the quality of care. Others claim that general training is adequate for most routine surgical procedures. The authors questioned whether there were differences in outcomes when general surgeons (GEN) operate on children and infants with common surgical conditions compared with the care of their pediatric surgical (PED) colleagues. METHODS: The authors retrospectively reviewed the Healthcare Investment Analysts North Carolina Information Network database to identify patients who underwent pyloromyotomy for congenital hypertrophic pyloric stenosis in North Carolina during the period from October 1995 through September 1998 (n = 780). Information obtained included demographics, insurance type, hospital, length of stay, total hospital charges, occurrence of mucosal perforation, and type of surgeon (general v pediatric). RESULTS: Of the 780 pyloromyotomies performed, 363 (48%) were performed by pediatric surgeons. Pediatric surgeons cared for more Medicaid patients than general surgeons (PED, 52% v GEN, 40%; P =.001). Infants treated by pediatric surgeons had a lower incidence of mucosal perforation (PED, 0.5% v GEN, 2.9%; P =.0015), which was associated with decreased overall total hospital charges (no perforation, $4,806 plus minus 79 v perforation, $6,592 plus minus 492; P =.002). When patients with uncomplicated pyloric stenosis were evaluated (96% of cases), those cared for by pediatric surgeons had lower total hospital charges (PED, $4,496 plus minus 95 v GEN, $5,121 plus minus 121; P =.0001) and shorter length of stay (PED, 2.7 plus minus 0.1 days v GEN, 3.1 plus minus 0.1 days; P =.01). CONCLUSIONS: In North Carolina, general surgeons treat more than half the patients who have pyloric stenosis, though fewer with Medicaid. The cost and incidence of mucosal perforation were increased in infants with pyloric stenosis when care was provided by general rather than pediatric surgeons. PMID- 11877648 TI - Congenital diaphragmatic hernia in 120 infants treated consecutively with permissive hypercapnea/spontaneous respiration/elective repair. AB - BACKGROUND/PURPOSE: Poor prognosis (approximately 50% survival rate and significant morbidity) traditionally has been associated with congenital diaphragmatic hernia (CDH). The authors reviewed a single institution experience and challenged conventional wisdom in the context of a care strategy based on permissive hypercapnea/spontaneous respiration/elective repair. METHODS: From August 1992 through February 2000, all infants with CDH and (1) respiratory distress requiring mechanical ventilation, (2) in-born or (3) transferred preoperatively within hours of birth are reported. All respiratory care strategy used permissive hypercapnea/spontaneous respiration and combined with elective repair. Arterial blood gas values and concomitant ventilator support were recorded. Outcome markers were (1) need for extracorporeal membrane oxygenation ECMO, (2) discharge to home, (3) supplemental oxygen need at discharge, and (4) influence of non-ECMO ancillary therapies (surfactant, nitric oxide, high frequency oscillatory ventilation). RESULTS: One hundred twenty consecutive infants were reviewed. Overall survival rate was 75.8%, but, excluding 18 of 120 not treated (6 lethal anomalies, 10 overwhelming pulmonary hypoplasia, 3 prerepair ECMO-related neurocomplications), 84.4% survived to discharge. A total of 67/120 were inborn. Non-ECMO ancillary treatments had no impact on survival rate. ECMO was used in 13.3%. Surgery was transabdominal; prosthetics were used in 7%. Tube thoracostomy was rare. Every inborn patient (n = 11) requiring a chest tube for pneumothorax died. Respiratory support before surgery was peak inspiratory pressure (PIP), 22, FIO(2),.43 with PaO(2), 66 torr; PaCO(2), 41 torr; and pH, 7.32. The survivors discharged on oxygen (n = 2) died at 4 and 7 months. CONCLUSIONS: The majority of infants with life-threatening CDH treated with permissive hypercapnea/spontaneous respiration/elective surgery survive to discharge with minimal pulmonary morbidity. PMID- 11877649 TI - Staged reconstruction of large congenital diaphragmatic defects with synthetic patch followed by reverse latissimus dorsi muscle. AB - BACKGROUND/PURPOSE: Synthetic repair of large congenital diaphragmatic defects (>90%) invariably will lead to recurrence, progressive chest wall deformity, and restrictive pulmonary disease. Staged reconstruction with living, growing tissue can help avoid these complications. METHODS: Between November 1995 and December 1999, 5 patients (median age, 25 months) with diaphragmatic agenesis underwent staged replacement with a reverse latissimus dorsi (RLD) flap. All required extracorporeal membrane oxygenation (ECMO) support at birth followed by synthetic patch (polytetrafluoroethylene or PTFE) diaphragm closure. Clinical evidence of patch disproportion, including recurrence (n = 3), chest wall deformity (n = 3), radiographic findings (n = 2), and restrictive respiratory patterns (n = 1), provided indication for replacement. The procedure involves removal of the original patch via thoracotomy followed by transposition of a RLD flap (based on the paraspinous and intercostal perforating vessels) into the defect through the bed of the 10th rib. Two patients underwent concomitant fundoplication. RESULTS: RLD flap was completed successfully in all 5 patients. Median length of stay after the procedure was 8 days. With a mean follow-up of 28 months (15 to 64 months), there have been no recurrences and no complications related to the procedure. Respiratory status and chest wall deformity have improved. There was neither evidence of paradoxical chest wall movement nor obvious limitation of the ipsilateral upper extremity on physical examination. The RLD flaps have grown proportionately to the children and remained at a stable level on chest x-ray. CONCLUSIONS: Staged reconstruction with patch closure followed by definitive reverse latissimus dorsi flap repair is a safe and highly effective treatment option in patients with diaphragmatic agenesis. A planned replacement should be considered in all patients with severe diaphragmatic hernias and patch closure before the development of thoracic complications. PMID- 11877650 TI - Management of late-stage parapneumonic empyema. AB - PURPOSE: Despite the reported value of early video-assisted thoracoscopic surgery (VATS) for empyema, many children are still referred to the surgeon late in the disease process. The authors wished to determine the optimal management strategy for this group of children. METHODS: Medical records of all children (n = 70) from 1990 to 2000 with late-presenting empyema (stage II or III) were reviewed. Patients were grouped as (G1) successful management with chest tube (CT), (G2) surgery after initial CT, (G3) thoracentesis followed by surgery, and (G4) surgery alone. RESULTS: There were no significant differences with respect to age, gender, pleural cultures or fluid analysis. Fifty-one (73%) patients required surgical intervention. Treatment using CT (G1, G2) or thoracentesis (G3) was associated with prolonged length of stay (LOS) when compared with surgery alone (G4; 12 v 8 days). For G2, G3, and G4, rapid clinical improvement and early discharge (6 days) was seen after surgery. For all surgery groups (G2, G3, G4), video-assisted thoracoscopic surgery (n = 19) was associated with a longer postoperative fever (4 v 2 days; P <.05), but a shorter total LOS (12 v 15 days; P <.05) when compared with open decortication (n = 32). CONCLUSIONS: Over 70% of children with late presenting empyema required surgery, including more than half of the children who received initial chest tube drainage. Delay in surgery was associated with more procedures, more radiographs, and an increased LOS. Despite later intervention, patients undergoing surgery as an initial approach had the shortest length of stay. Early surgical intervention is indicated for most children referred with established empyema. PMID- 11877651 TI - All-terrain vehicle and bicycle crashes in children: epidemiology and comparison of injury severity. AB - BACKGROUND/PURPOSE: Despite statements by the American Academy of Pediatrics (AAP) and the US Consumer Product Safety Commission (CPSC) against the use of all terrain vehicles (ATVs) by children under the age of 16 years, nearly half of ATV related injuries and over 35% of all ATV-related deaths continue to occur in this age group. Because ATV and bicycle crashes have been associated with serious injury in children, the authors compared the demographics, mechanism of injury, injury severity, and outcome of children with ATV- and bicycle-related injuries. Further, the authors sought to identify whether ATV-related injuries elicited changes in risk-taking behavior. METHODS: A retrospective, comparative analysis of 109 children admitted for ATV-related injuries and 994 children admitted for bicycle-related injuries to a level 1 pediatric trauma center between January 1991 and June 2000 was performed. A phone survey was conducted to determine self reported changes in safety behaviors or use patterns after ATV injury. RESULTS: Mean age was 11.1 plus minus 3.5 years (range, 2 to 18 years) for ATV crashes versus 9.4 plus minus 3.3 years (range, 1 to 17 years) for bicycle crashes (P <.05). Ninety-three percent of ATV crashes occurred in children less than 16 years of age; 31% in children less-than-or-equal10 years of age; and 7% in children less-than-or-equal5 years of age. Male-to-female ratio was about 3:1 for both groups. White race accounted for 97% of ATV injuries compared with 79% of bicycle injuries (P <.05). Falls from ATVs or bicycles were the most common mechanism of injury (41% v 59%, respectively). Collisions with motor vehicles were more common for bicyclists (32% v 10%), whereas collisions with stationary objects were more common among ATV riders (27% v 9%). Sixteen percent of ATV crashes were caused by a roll-over mechanism. Mean injury severity score (ISS) were significantly higher for victims of ATV crashes (8.3 ATV v 6.7 bicycle; P <.05). ATV-related trauma was associated with multiple injuries, more operative interventions, and longer hospital stays. Location and distribution of injuries were similar for both groups. Helmet use was low in both groups but higher for ATV riders (23% v 8%; P <.5). Mortality rate was similar for both groups (0.9% for ATV riders v 0.7% for bicyclists). There was a 39% response for the phone survey post-ATV injury. Twenty-three of 43 (53%) respondents owned the ATV, and 70% of these received safety information at the time of purchase. However, only 14% of injured riders received any formal training before riding ATVs. Postinjury, 60% of children continued to ride, although 42% reported decreased riding time. Fifty-four percent of children reportedly wore helmets preinjury, and there were no changes in helmet usage postinjury. There were no differences in pre- and postinjury parental supervision (61% v 65%). CONCLUSIONS: Both ATV and bicycle-related injuries occur predominantly in boys, but ATV victims are older and almost all are white. Almost all ATV injuries occurred in children under the age of 16 years. Although both ATV and bicycle crashes cause severe injuries in children, injury severity is higher for ATV crashes in terms of multiple injuries, need for operative intervention, and longer length of stay. Despite severe injuries, the majority of children injured by ATVs continue to ride, albeit fewer hours per day, and safety behaviors are unaltered. These data reinforce the current AAP stance that legislation prohibiting the use of ATVs in children under the age of 16 years without a valid driver's license should be pursued and enforced aggressively. PMID- 11877652 TI - Reimaging in pediatric neurotrauma: factors associated with progression of intracranial injury. AB - PURPOSE: The purpose of this study was to characterize the radiologic changes that are seen in the first 24 to 48 hours after head injury and to correlate those changes with clinical findings, to determine which children are at greatest risk for progression of their neurologic injury. METHODS: The authors identified 104 children (less-than-or-equal17 years of age) who had a second computed tomography (CT) scan of the head within 24 to 48 hours of admission. CT scans were evaluated systematically in a blinded fashion. Mechanism of injury, findings on physical examination, therapeutic measures, and changes in management were recorded from hospital medical records. The 50 children whose second CT scan showed progression of injury were compared with the 54 patients whose intracranial injuries were unchanged or improved on their second CT. RESULTS: Twenty-six percent of patients (13 of 50) with radiographic progression of injury had an admission Glasgow coma score of 15. Progression of injury was more common, however, in patients with lower Glasgow coma scores, averaging 9 on admission and 10 at the time of the second CT. Progression of injury also was more common if the initial head CT showed 3 or more intracranial injuries, mass effect, intraventricular hemorrhage, or an epidural hematoma. CONCLUSIONS: Children with an intracranial injury identified on their initial head CT scan should undergo a second scan 24 hours after injury, especially if the initial CT shows 3 or more intracranial injuries, mass effect, intraventricular hemorrhage, or an epidural hematoma. . PMID- 11877653 TI - Utility of radiographic hepatic injury grade in predicting outcome for children after blunt abdominal trauma. AB - BACKGROUND/PURPOSE: The American Association for the Surgery of Trauma (AAST) liver injury grading system has been adopted in the management of pediatric hepatic injuries. However, the usefulness of this grading system in children remains undefined. The authors, therefore, examined the validity of AAST grading in the management of pediatric blunt liver injury. METHODS: The authors identified 152 patients, ages 0 to 18 years, with blunt hepatic injury treated at our pediatric trauma center between 1995 and 2000. Radiographic AAST grade was assigned retrospectively by 2 radiologists. Variables analyzed included age, gender, Glasgow Coma Score (GCS), Injury Severity Score (ISS), and associated injuries. Outcome measures were mortality rate and length of stay (LOS). Statistical analysis was performed using analysis of variance or linear regression. RESULTS: Computed tomography (CT) scans were available for 95 patients. Radiographic injury grade did not correlate with mortality rate, hospital LOS or intensive care unit (ICU) LOS. ISS and associated injury, but not age, gender, or GCS, were predictive of LOS. Grade did not correlate with mortality rate, ICU LOS, or hospital LOS. CONCLUSIONS: Radiographic liver grading does not predict outcome reliably in children and should not be the main parameter utilized to guide clinical decision making. A role for scoring systems that utilize factors such as associated injuries and ISS is indicated. PMID- 11877654 TI - Intestinal permeability and bacterial translocation are uncoupled after small bowel resection. AB - BACKGROUND/PURPOSE: Gut barrier failure and bacterial translocation have been proposed to cause infection and sepsis in patients with the short bowel syndrome. This study tested the hypothesis that permeability is increased in the adapting remnant ileum after massive small bowel resection (SBR). METHODS: Male ICR mice underwent a 50% proximal SBR or sham operation. At 3, 7, and 14 days, the ileum was mounted in an Ussing chamber. Mucosal-to-serosal flux of low (dextran) and high (horseradish peroxidase; HRP) molecular weight markers was determined. Additionally, bacterial translocation was measured by culturing blood, mesenteric lymph nodes, liver, and spleen at 3 and 14 days after SBR or sham operation. RESULTS: Permeability to dextran was reduced immediately after SBR but was no different at later time-points. HRP permeability was no different at any time point. Translocation of Gram-negative bacteria to the mesenteric lymph nodes and liver was more frequent in the SBR animals 3 and 14 days postoperatively. CONCLUSIONS: Intestinal permeability to macromolecules is not increased after massive SBR, but the rate of translocation to the mesenteric lymph nodes and liver is elevated. This suggests that the mechanism for bacterial translocation after SBR does not involve alterations in gut permeability. PMID- 11877655 TI - Laparoscopic inguinal herniorrhaphy in children: a three-center experience with 933 repairs. AB - BACKGROUND/PURPOSE: Laparoscopic inguinal herniorrhaphy has been introduced recently as an alternative to conventional open repair in children. This study was undertaken to evaluate the safety, efficacy, and reproducibility of this minimally invasive approach. METHODS: A total of 933 laparoscopic inguinal herniorrhaphies were performed on 666 children (597 boys and 69 girls), ranging in age from 3 weeks to 14 years (median, 3.2 years). A 5-mm laparoscope was placed through an umbilical incision, and two 2-mm or 3-mm needle drivers were inserted through the lateral abdominal wall. The neck of the sac was closed with a 4-0 monofilament suture. The needle was inserted directly through the abdominal wall, and removed together with the trocar. Only the umbilical fascia was closed with an absorbable suture. No skin sutures were applied. RESULTS: A total of 911 indirect inguinal hernia sacs were closed (337 right, 172 left, 402 bilateral) and 22 direct inguinal hernias were repaired (14 boys, 3 girls; 11 right, 3 left, 4 bilateral). The median operating time was 22 minutes (range, unilateral, 7 to 45 min; bilateral, 9 to 51 min). With experience, this time gradually decreased. There were no intraoperative complications. The contralateral asymptomatic processus was unexpectedly open on the left side in 137 of the boys (23%) and 10 of the girls (15%), and on the right side in 131 of the boys (22%) and 21 of the girls (32%). In 16% of the children, the final procedure was modified on the basis of the anatomic findings. No hernia was found in 13 children (1.9%). The recurrence rate was 3.4% (follow-up time ranged from 2 months to 7 years). Hydroceles were observed in 4 children, and a subtle change in testicular position and size was noted in one boy. CONCLUSIONS: Laparoscopic inguinal repair in children proved safe and reproducible, although the recurrence rate was slightly higher than with the open approach. However, laparoscopy allows easy and precise identification of the type of defect and its correction. In this series, the incidence of direct inguinal hernias was higher, and the incidence of a patent contralateral processus vaginalis was lower than previously reported. PMID- 11877656 TI - Balloon catheter dilatation in children with congenital and acquired esophageal anomalies. AB - BACKGROUND/PURPOSE: Esophageal stricture requiring dilatation often is the sequelae in children with a history of congenital or acquired esophageal anomalies. Balloon catheter dilatation (BCD) for esophageal stricture is well established in adults, yet its role in children is less well described. The authors report their initial experience using BCD in children with esophageal stricture, assessing outcome. METHODS: Between January 1994, and December 1998, 20 children (age range, 17 days to 12 years) underwent BCD for esophageal strictures and were divided into 2 etiology groups (A and B). (A, n = 10) included congenital anomalies (esophageal atresia, 10), and (B, n = 10) included acquired anomalies (caustic ingestion, 7; tight fundoplication, 2; peptic stricture, 1). BCD was performed in all cases under conscious sedation with a balloon catheter (6 to 25 mm) with fluoroscopy. Outcome parameters included number of dilatations; complications, ie, perforation; and success or failure of procedure. Success was defined as increasing intervals of age-appropriate food tolerance between dilatations, and failure was defined as abandonment of dilatation in favor of surgical intervention. Follow-up for A and B ranged from 6 to 42 months. RESULTS: Seventeen of 20 (85.0%) underwent successful BCD. In A, 10 of 10 (100%) had complete resolution of symptoms with follow-up ranging from 6 to 42 months versus 7 of 10 (70.0%) in B with follow-up of 6.5 to 36 months. In A, number of dilatations ranged from 1 to 4 over an average period of 2 months. In B, number of dilatations ranged from 3 to 40 over periods ranging from 2 to 30 months. All 3 failures occurred in children with caustic ingestion, with 1 child suffering perforation requiring urgent surgical intervention. CONCLUSIONS: Balloon catheter dilatation can be applied safely and effectively to children in whom esophageal strictures develop after repair of esophageal atresia. However, its use in children with acquired esophageal anomalies, notably caustic injury, is associated with higher complication and failure rates, necessitating greater caution and lower expectations. PMID- 11877657 TI - Application of the principles of myotomy and strictureplasty for treatment of esophageal strictures. AB - BACKGROUND/PURPOSE: Strictures of the esophagus in the pediatric population may be congenital or acquired. Regardless of the etiology, the treatment goal is to relieve the symptoms and allow patients to eat normally. The cornerstone of nonoperative management is repeated esophageal dilatations. However, when nonoperative management fails, operative intervention becomes necessary. In this report the authors present 4 cases of severe upper esophageal strictures managed by applying the principles of myotomy and strictureplasty. METHODS: Retrospective review of 4 patients undergoing esophageal stricturotomy from January 1, 1993 to January 1, 2000 was conducted at Childrens Hospital Los Angeles, with a mean follow-up period of 5 years. RESULTS: Three of the 4 patients reported in this report are doing well and have not required any further surgical intervention. One patient in whom additional strictures developed has had a microvascularized free jejunal graft and also is doing well. CONCLUSIONS: Currently accepted surgical management of strictures includes resection of the affected segment with end-to-end anastomosis or esophageal replacement. The authors propose that before embarking on a long and technically hazardous operation, consideration should be given to stricturotomy. In cases of isolated strictures, this may be the preferred approach. PMID- 11877658 TI - Fundoplication and gastrostomy versus image-guided gastrojejunal tube for enteral feeding in neurologically impaired children with gastroesophageal reflux. AB - BACKGROUND: Neurologically impaired children with gastroesophageal reflux (GER) usually are treated with a fundoplication and gastrostomy (FG); however, this approach is associated with a high rate of complications and morbidity. The authors evaluated the image-guided gastrojejunal tube (GJ) as an alternative approach for this group of patients. METHODS: A retrospective review of 111 neurologically impaired patients with gastroesophageal reflux was performed. Patients underwent either FG (n = 63) or GJ (n = 48). All FGs were performed using an open technique by a pediatric surgeon, and all GJ tubes were placed by an interventional radiologist. RESULTS: The 2 groups were similar with respect to diagnosis, age, sex and indication for feeding tube. Patients in the GJ group were followed up for an average of 3.11 years, and those in the FG group for 5.71 years. The groups did not differ statistically with respect to most complications (bleeding, peritonitis, aspiration pneumonia, recurrent gastroesophageal reflux [GER], wound infection, failure to thrive, and death), subsequent GER related admissions, or cost. Children in the GJ group were more likely to continue taking antireflux medication after the procedure (P <.05). Also, there was a trend for GJ patients to have an increased incidence of bowel obstruction or intussusception (20.8% v 7.9%). Of the FG patients 36.5% experienced retching, and 12.7% experienced dysphagia. Eighty-five percent of patients in the GJ group experienced GJ tube-specific complications (breakage, blockage, dislodgment), and GJ tube manipulations were required an average of 1.68 times per year follow-up. Nine patients (14.3%) in the FG group had wrap failure, with 7 (11.1%) of these children requiring repeat fundoplication. In the GJ group, 8.3% of patients went on to require a fundoplication for persistent problems. A total of 14.5% of GJ patients had their tube removed by the end of the follow-up period because they no longer needed the tube for feeding. CONCLUSIONS: Image-guided gastrojejunal tubes are a reasonable alternative to fundoplication and gastrostomy for neurologically impaired children with GER. The majority can be inserted without general anesthesia. This technique failed in only 8.3% patients, and they subsequently required fundoplication. A total of 14.5% of GJ patients showed some spontaneous improvement and had their feeding tube removed. Each approach, however, still is associated with a significant complication rate. A randomized prospective study comparing these 2 approaches is needed. PMID- 11877659 TI - Comparison of minimally invasive and modified Ravitch pectus excavatum repair. AB - BACKGROUND/PURPOSE: Minimally invasive repair of pectus excavatum (MIRPE) has gained wide acceptance during the last 4 years. This study compares, retrospectively, the experience at 2 large hospitals, 1 using MIRPE and the other a modified Ravitch repair (MRR). METHODS: From 1996 to 2000, 68 PE patients underwent MIRPE at one hospital, and 139 underwent MRR at another hospital. Ages ranged from 5 to 19 years (mean, 12) for MIRPE, and 3 to 51 years (mean, 17.3) for MRR. The mean pectus severity index was 4.2 for MIRPE and 4.9 for MRR (normal, 2.5). RESULTS: There were no deaths after MIRPE or MRR. Complications included 6 reoperations for MIRPE and none for MRR. There were 8 rehospitalizations for MIRPE and none for MRR. Ninety percent of MIRPE complications occurred in the first 25 cases. The mean blood loss was under 90 mL for both MIRPE and MRR. Mean operating time was 75 minutes for MIRPE and 212 minutes for MRR. Ninety-six percent of MIRPE patients and no MRR patients had epidurals. Intravenous analgesics averaged 5 days for MIRPE and 1.7 days for MRR. Mean hospitalization was 6.5 days for MIRPE and 2.9 days for MRR. Mean time before return to work or school was 18 days for MIRPE and 12 days for MRR. The sternal bar was removed from 107 of 139 MRR patients (mean time, 19 minutes) and 18 of 68 MIRPE patients (mean time, 25 minutes). CONCLUSIONS: Both MIRPE and MRR provide excellent clinical results. MRR has a longer operating time but decreased hospital stay, complication rate, and use of pain medications. Attention to technical operative details and surgeon's experience are essential for optimal results using both techniques. PMID- 11877660 TI - The EXIT procedure: experience and outcome in 31 cases. AB - BACKGROUND: The EXIT (ex utero intrapartum treatment) procedure, although initially designed for reversal of tracheal occlusion in fetuses with congenital diaphragmatic hernias (CDH), has been adapted to treat a variety of fetal conditions. METHODS: A retrospective chart review of all consecutive EXIT procedures since 1996 was conducted. RESULTS: Thirty-one women underwent the EXIT procedure, with an average maternal age of 29 years (range, 20 to 38), and average gestational age of 34 weeks (range, 29 to 40). The indication was airway obstruction from fetal neck mass in 13, and reversal of tracheal occlusion from in utero clipping in 13. Singular indications included an EXIT-to-ECMO (extracorporeal membrane oxygenation) procedure for a fetus with CDH and a cardiac defect (n = 1), congenital high airway obstruction syndrome (CHAOS, n = 1), resection of a very large congenital cystic adenomatoid malformation of the lung (CCAM) on uteroplacental bypass (n = 1), unilateral pulmonary agenesis (n = 1), and thoracoomphalopagus conjoined twins. The mean duration on uteroplacental bypass (from uterine incision to umbilical cord clamping) was 30.3 plus minus 14.7 minutes (range, 8 to 66). No fetus experienced hemodynamic instability during uteroplacental bypass as recorded by fetal heart rate (FHR), pulse oximeter, and fetal echocardiography, except for one instance of reversible bradycardia from umbilical cord compression. The mean FHR and fetal saturation were 153.0 plus minus 38.5 beats per minute and 71.2% plus minus 19.9%, respectively. Five fetuses required a tracheostomy. Only 1 death occurred during an EXIT procedure because of inability to secure the airway secondary to extensive involvement by a lymphangioma. The average cord pH and pCO(2) were, respectively, 7.20 plus minus 0.11 and 63.2 plus minus 14.6. Two maternal complications occurred: bleeding from a hysterotomy site and dehiscence of an old hysterotomy scar noticed at a subsequent cesarean section. The average maternal blood loss was 848.3 plus minus 574.1 mL. CONCLUSION: The EXIT procedure was used successfully to ensure uteroplacental gas exchange and fetal hemodynamic stability during a variety of surgical procedures performed to secure the fetal airway or ensure successful transition to postnatal environment. PMID- 11877661 TI - Economic implications of current surgical management of gastroesophageal reflux disease. AB - BACKGROUND/PURPOSE: Surgical management of gastroesophageal reflux disease in children has evolved with the development of laparoscopy. Because concerns persist regarding increased costs associated with this technique, the authors studied the economic parameters of antireflux surgery at their institution. METHODS: Seventy-eight patients undergoing either laparoscopic or open fundoplication were studied retrospectively between June 1998 and June 2000 comparing average operating room costs, total inpatient costs, and length of stay. Univariate comparisons were performed using Student's t test, and multivariate analysis was performed using multiple linear regression. RESULTS: Univariate analysis showed that patients receiving the laparoscopic procedure had significantly shorter inpatient stays (2.4 v. 3.96 days; P =.004) than those receiving open procedures. Average operating room costs were similar (laparoscopic, $2,611; open, $2,162; P =.237), but total costs for the laparoscopic procedure were lower ($4,484 v $5,129; P =.006). Multivariate analysis results suggested that in addition to procedure type, patients who required an intensive care unit admission incurred $6,595 in additional total costs (P <.0001) and 4.8 additional hospital days (P <.0001). After controlling for other variables, the laparoscopic procedure did not significantly reduce total hospital costs ($447; P =.192) but was associated with a significant decrease in length of stay of 1.3 days (P <.0001). CONCLUSION: These results suggest that laparoscopic procedures are comparable with open operations in terms of operative costs and that other factors are important determinants of the costs associated with antireflux surgery in children. PMID- 11877662 TI - Lessons learned from a failed multi-institutional randomized controlled study. AB - BACKGROUND/PURPOSE: In 1996, the Surgical Sections of the Children's Cancer Group (CCG) and the Pediatric Oncology Group (POG) received National Cancer Institute funding to conduct a prospective, randomized, controlled, surgeon-directed study to evaluate the role of minimally invasive surgery (MIS) in children with cancer. Because of lack of patient accrual, the study was closed in 1998. The purpose of this study is to evaluate and describe those factors that impacted on study failure to ensure future successful clinical trials. METHODS: One hundred forty surgeons representing the surgical membership of CCG and POG as well as 111 institutions within CCG and POG were asked to complete a questionnaire about the failed clinical trial. The questionnaire focused on study objectives, organization, and institutional review board (IRB) submission. It also examined the surgeon's ability to perform the minimal access operation, the influence of the pediatric oncologist, and the existence of preconceived biases by surgeons, oncologists, and families. Statistical analysis was performed as appropriate. RESULTS: Eighty-six of 140 (62%) surgeons responded to the questionnaire. Only 23% of the potential protocols were submitted for IRB approval. Of responding surgeons, 39% were not actively performing MIS when the study opened. A surgeon's support of the study was directly related to when the surgeon received the protocols (P <.001) and whether the participating surgeon was actively participating in MIS (P <.016). The oncologist's knowledge and support of the study affected IRB submission and approval (P <.02) and was influenced by whether MIS was practiced at the institution (P <.05). The majority of responding surgeons believed the experimental question was relevant (P <.05). However, responding surgeons believed that a preconceived bias existed within both their local surgical and oncology communities favoring a particular surgical approach (P <.001), but this bias did not extend to the families (P >.05). CONCLUSION: The study failed because of lack of accrual for a variety of reasons: failure to submit to the institution's IRB, lack of surgical expertise with MIS procedures, and preconceived surgeon bias toward either an endoscopic or traditional open approach. PMID- 11877663 TI - Experience and modification update for the minimally invasive Nuss technique for pectus excavatum repair in 303 patients. AB - PURPOSE: The aim of this study is to review the new technical modifications and results of 303 patients who have had pectus excavatum repair utilizing the minimally invasive technique. METHODS: A retrospective chart review was conducted of 303 patients undergoing minimally invasive pectus repair from 1987 through August 2000. Since 1997, a standardized treatment pathway was implemented, and 261 of the 303 patients have been treated on this pathway. Preoperative evaluation included computed tomography (CT) scan, pulmonary function tests (PFT), and cardiac evaluations with electrocardiogram (EKG) and echocardiogram. Indications for operation included at least 2 of the following: progression of the deformity, exercise intolerance or restrictive disease on PFT, Haller CT index greater than 3.2, mitral valve prolapse (MVP), or cardiac compression. Technical and design modifications since 1998 have included routine thoracoscopy, the use of an introducer/dissector for creating the substernal tunnel and elevating the sternum, and routine use of a wired lateral stabilizer to prevent bar displacement. The bar is removed as an outpatient procedure in 2 to 4 years. RESULTS: In 303 patients undergoing minimally invasive pectus repairs, single bars were used in 87% and double in 13%. Lateral stabilizers were applied in 70% of patients and were wired for further stability in 65%. Bar shifts before the use of stabilizers were 15%, which decreased to 6% after stabilizers were placed and 5% with a wired stabilizer. Excellent results were noted in 85% with failure in only 1 patient. Complications included pneumothorax with spontaneous resolution in half of the patients and pericarditis in 7. CONCLUSIONS: The minimally invasive technique has evolved into an effective method of pectus excavatum repair. Modifications of the technique have reduced complications. Long term results continue to be excellent. PMID- 11877664 TI - Open lung biopsy in pediatric bone marrow transplant patients. AB - PURPOSE: The aim of this study was to evaluate the clinical benefits of open lung biopsy in the diagnosis and treatment of pulmonary infiltrates in children who have undergone bone marrow transplantation. METHODS: The authors retrospectively reviewed the medical records of all patients in whom pulmonary infiltrates developed within 6 months after bone marrow transplantation. Of 528 patients who received bone marrow transplants (313 allogeneic, 215 autologous) at St Jude Children's Research Hospital between June 1991 and December 1998, 83 (16%) had radiographic evidence of pulmonary infiltrates after the procedure. Of these, 43 (52%) underwent bronchoalveolar lavage (BAL), 19 (23%) underwent open lung biopsy (OLB), 6 (7%) underwent needle biopsy, and 5 (6%) underwent transbronchial biopsy; 21 received medical therapy alone. The authors evaluated the outcome, culture results, histopathologic findings, radiographic findings, and clinical features of those who underwent OLB. RESULTS: The 19 patients ranged in age from 0.9 to 19.8 years (median, 11.4 years). Histopathologic studies indicated an infectious process in 6 patients (30%), bronchiolitis obliterans organizing pneumonia (BOOP) in 5 (26%), interstitial pneumonitis (IP) in 4 (21%), gangliosidosis in 1, and lymphocytic infiltrate in 1. Although the clinical plan was changed on the basis of the histopathologic diagnosis for 17 of the 19 patients (90%), improvement in outcome was seen in only 8 (47.5%) of these patients. Postoperative morbidity (30 days) was 47% and included prolonged intubation (7 patients), pneumothorax (2 patients), and pleural effusion (1 patient). The 30-day survival rate was 63.2% plus minus 10.6%. No patient with multisystem organ failure (MSOF), ventilator dependence, or a postoperative complication survived after OLB. CONCLUSIONS: Histopathologic analysis of OLB specimens is very accurate in determining the cause of pulmonary infiltrates in pediatric patients who have undergone BMT, but OLB may not improve patient outcome. Because the postoperative morbidity and mortality rates associated with OLB are high, careful patient selection is necessary. The mortality rates of patients with MSOF or ventilator dependence are particularly high; therefore, less-invasive alternatives for diagnosis of pulmonary lesions should be considered before OLB is performed. PMID- 11877665 TI - Compliance with evidence-based guidelines in children with isolated spleen or liver injury: a prospective study. AB - PURPOSE: This prospective study is designed to disseminate and apply 5 previously defined, evidence-based guidelines in children with isolated spleen or liver injuries. The desired outcomes are to standardize treatment, promote consensus, and utilize resources in a safe and efficient manner. METHODS: Guidelines on intensive care unit (ICU) stay, hospital stay, imaging, and physical activity restriction for children with isolated spleen or liver injuries were defined by retrospective analysis of 832 children treated nonoperatively at 32 centers from 1995 to 1997. The guidelines were based on severity of injury by computed tomography (CT) grade. These guidelines were applied prospectively in 312 children treated nonoperatively at 16 centers from 1998 to 2000. Compliance was analyzed for age, gender, organ injured, and CT grade. All patients underwent follow-up for 4 months. In addition, the authors compared ICU stay, hospital stay, follow-up imaging, and interval of activity restriction in the retrospective historical group (1995 through 1997) with the current prospective group (1998 through 2000). RESULTS: Specific guideline compliance was 81% for ICU stay, 82% for hospital stay, 87% for follow-up imaging, and 78% for interval of activity restriction. There was a significant improvement in compliance from year 1 to year 2 for ICU stay (77% v 88%; P <.02) and interval of activity restriction (73% v 87%; P <.01). Compliance with proposed intervals for activity restriction was greater for those with less severe injuries (87% in grades I and II v 72% in grades III and IV; P <.01). There were no differences in compliance by age, gender, or organ injured. Deviation from guidelines was surgeon's choice in 90% and patient related in 10%. Six (1.9%) patients were readmitted, although none required operation. Compared with the previously studied 832 patients, the 312 patients who had prospective application of the proposed guidelines had a significant reduction in ICU stay (P <.0001), hospital stay (P <.0006), follow-up imaging (P <.0001), and interval of physical activity restriction (P <.05) within each grade of injury. CONCLUSIONS: Prospective application of specific treatment guidelines based on injury severity has resulted in conformity in patient management, improved utilization of resources, and validation of guideline safety. Significant reduction of ICU stay, hospital stay, follow-up imaging, and length of activity restriction has been achieved without adverse sequelae when compared with our retrospective database. PMID- 11877666 TI - Interleukin-11 enhances intestinal absorptive function after ischemia-reperfusion injury. AB - BACKGROUND/PURPOSE: Interleukin-11 (IL-11) is a multifunctional cytokine that has been shown to improve small bowel adaptation and enhance cellular recovery after bowel ischemia. This study was designed to examine the effects of systemic IL-11 on small bowel absorptive function in a rat model of intestinal ischemia and reperfusion (IR) injury. METHODS: Sprague-Dawley rats underwent placement of a venous catheter connected to an osmotic pump, which delivered its contents over 3 days. Rats were divided into 3 groups: sham operation/systemic saline; 30-minute superior mesenteric artery occlusion/systemic saline; superior mesenteric artery occlusion/systemic IL-11, 750 microgram/kg/d. After the infusion, (14)C-galactose or (14)C-glycine absorption was measured using an in vivo, recirculation technique. Statistical significance was determined using analysis of variance. RESULTS: In control rats, 30 minutes of IR decreased absorption of galactose from 2.62 to 2.02 micromoles/cm(2) (P <.01), and glycine from 2.79 to 1.72 micromoles/cm(2) (P <.01). Rats treated with systemic IL-11 showed improved absorption of galactose of 2.39 micromoles/cm(2) (P <.05), and glycine at 2.21 micromoles/cm(2) (P <.05). Mucosal DNA content was reduced significantly from 7.37 to 5.61 microgram DNA/mg by IR (P <.01). IL-11 treatment did not significantly alter DNA content during this period. CONCLUSIONS: These data show that 30 minutes of intestinal IR significantly decreases intestinal absorptive function in this animal model. When compared with untreated control animals, administration of systemic IL-11 significantly increased the absorption of carbohydrate and amino acid in rats recovering from mesenteric IR. PMID- 11877667 TI - Long-term nutritional outcome after pediatric intestinal transplantation. AB - BACKGROUND/PURPOSE: The aim of this study was to describe the long-term nutritional status of a large population of children after intestinal transplantation and to identify factors associated with nutritional outcomes. METHODS: Longitudinal anthropometric data are maintained in a database registry for all patients referred to our Intestinal Care Center (ICC). Z-scores for weight and height were calculated biannually over a maximum of 2 years, and associations between baseline and follow-up laboratory measures and growth were evaluated for patients greater than 6 months post intestinal transplant. RESULTS: Since the inception of the ICC in December 1996, 24 pediatric patients (18 boys, 18 white) received an isolated small bowel or small bowel/liver transplant (median age, 3.2 years). The majority of cases (75%) had been diagnosed with surgical short bowel syndrome and were dependent on total parenteral nutrition (TPN) at the time of transplant. Of the 23 patients who survived the initial postoperative period, 87% were weaned from TPN to an amino-acid or peptide-based enteral formula or solid food within 3 months. A positive trend in z-scores for weight and height/length was observed in only 30% and 26% of patients, respectively, during the follow-up period. Although mean albumin levels increased significantly from 2.8 to 3.1 mg/dl by 6 months posttransplant (P <.01) no difference in alkaline phosphatase was found over time. Steroid doses were weaned within 3 to 4 months after transplantation but not discontinued. The cumulative survival rate was 91% at 1 year and 86% at 2 years posttransplant, whereas those weaned from TPN achieved 100% and 94% survival, respectively. CONCLUSIONS: Attainment of positive linear growth remains a challenge in the pediatric transplant population despite successful liberation from TPN, protein anabolism, and high survival rates. Further investigation into alternative methods of nutritional evaluation and manipulation as well as the use of growth factors to enhance the growth process need to be investigated. PMID- 11877668 TI - Nutritional outcome and growth of children after intestinal transplantation. AB - PURPOSE: The aim of this study was to determine nutritional outcome and growth in children after successful intestinal transplantation. METHODS: Case-record review was conducted of all children who underwent intestinal transplantation at a single center and retained their grafts for at least 1 year. Supplementary data were obtained from outpatient charts and computerized database. RESULTS: Forty seven intestinal transplants were carried out in 46 children. There were 19 isolated small bowel and 29 combined liver--small bowel transplant procedures. Median age at transplantation was 3.7 years (range, 0.4 to 16.6 years), and median graft survival time was 1,084 days (range, 368 to 3308 days). Nine patients died, and there were 11 graft losses, including those of the nonsurvivors. All survivors with functioning grafts receive all of their calories via the enteral route. There was significant inhibition of linear growth at the time of transplant in the majority of recipients. After successful transplantation, pretransplant growth velocity appeared to be maintained, but there was no evidence of catch-up growth. CONCLUSIONS: Intestinal transplantation allows an opportunity for full enteral feedings to be established. There is evidence of severe inhibition of linear growth at the time of transplantation with no evidence of catch-up after transplantation. PMID- 11877669 TI - VEGF upregulates Bcl-2 expression and is associated with decreased apoptosis in neuroblastoma cells. AB - BACKGROUND/PURPOSE: Both the expression of Bcl-2 and the amount of vascular endothelial growth factor (VEGF) are increased in neuroblastoma cells cocultured with hepatocytes. The authors hypothesize that VEGF upregulates Bcl-2 expression by the neuroblastoma cells and protects them from apoptotic stimuli. METHODS: To determine whether VEGF will induce Bcl-2 expression in neuroblastoma cells, the cells are plated with standard media (control) or media supplemented with VEGF. After 24 hours, Bcl-2 expression is measured. To determine whether VEGF protects neuroblastoma cells from apoptosis, the cells are subjected to tumor necrosis factor alpha (TNF-alpha) or serum starvation to induce apoptosis either with or without VEGF added to the culture media. The cells are collected and apoptosis measured using the deoxynucleotidyltransferase-mediated dUTP neck end labeling (TUNEL) method. RESULTS: VEGF increases Bcl-2 expression by 33% over cells cultured in standard media. Serum starving the tumor cells or adding TNF-alpha significantly increases the percentage of apoptotic cells. The addition of VEGF significantly protects the neuroblastoma cells from the apoptotic effects of both serum starvation and TNF-alpha. CONCLUSIONS: VEGF increases the expression of Bcl 2 and also abrogates TNF-alpha and serum starvation-induced apoptosis in neuroblastoma cells in vitro. VEGF may promote neuroblastoma survival not only through angiogenesis, but also by altering apoptosis and its regulating proteins. PMID- 11877670 TI - Insulin-like growth factor-I protects neuroblastoma against starvation-induced apoptosis and is associated with increased Bcl-2 expression. AB - BACKGROUND/PURPOSE: Aggressive neuroblastomas avoid apoptosis and have increased expression of the antiapoptotic protein, Bcl-2. Insulin-like growth factor-I (IGF I) is mitogenic and may promote tumor survival by inhibiting apoptosis. The authors hypothesize that IGF-I may protect neuroblastoma cells from apoptosis by upregulating their Bcl-2 expression. METHODS: Human neuroblastoma cells (IMR-32) are cultured, and 3 experimental groups are established: 1 group with cells cultured in standard growth media (control), 1 with cells grown in serum-depleted media (starvation), and 1 with neuroblastoma cells cultured in starvation media plus IGF-I. The cells are harvested at 14 and 24 hours, and cytospin slides are made. Bcl-2 expression is measured by immunohistochemistry. Apoptosis is detected with the TUNEL method. RESULTS: Bcl-2 expression is decreased 90% in the serum starved neuroblastoma cells. In addition, apoptosis is 150 times higher in the starved neuroblastoma cells. These changes are abrogated by the addition of IGF I, where apoptosis is decreased 50% and Bcl-2 is 14-fold higher in the IGF-I treated group. These changes are most apparent at 24 hours. CONCLUSIONS: IGF-I protects neuroblastoma cells from apoptosis and increases Bcl-2 expression. Growth factors may have a direct role in promoting tumorigenesis by inducing the expression of antiapoptotic proteins by the tumor. PMID- 11877671 TI - Desktop and mobile software development for surgical practice. AB - PURPOSE: The aim of this study was to develop a mobile software solution that will merge patient data with a centralized patient database, aid surgical decision making, and document clinical services. METHODS: iIncise is a computerized, menu-driven database that was developed for real-time entry of historical, clinical, and laboratory data. Within the process, examination data, testing rationale, and test results are captured. AAST staging criteria are presented during data entry to describe and classify organ injuries accurately. A legible, time-stamped, conclusive report can be generated for printed or electronic entry into the medical record. The handheld version of the database runs on a Compaq iPAQ Pocket PC in either stand-alone mode or via LAN or WAN through the Internet. Microsoft Information Internet Server 5.0 provides data transaction services to Microsoft SQL Server 2000 to merge multiuser replicate data between the back ends stored on the desktop and handheld devices. RESULTS: The printed report provides superior documentation for comprehensive evaluation and management services, including history and physical examination, documentation of medical decision making, appropriateness of diagnostic/or therapeutic services, and coordination of care in accordance with Health Care Financing Administration (HCFA) regulations. CONCLUSIONS: Rapid data entry, ease of use, and appropriate documentation of detailed and legible patient visits and procedural notes are early benefits. Wireless data transmission between handheld devices and the desktop database provides the required speed, flexibility, and multitasking environment necessary for the mobile surgeon. PMID- 11877672 TI - Effects of tezosentan, a dual endothelin receptor antagonist, on the cardiovascular and renal systems of neonatal piglets during endotoxic shock. AB - BACKGROUND/PURPOSE: Endothelin is a potent mediator of the cardiovascular and renal systems. Studies have found that endothelin has an important role in regulating cardiac function and renal perfusion in neonates who are suffering from endotoxic shock. The authors believe that blockade of the endothelin response during endotoxemia will have a beneficial effect on neonatal cardiac and renal functions. In this study the authors have examined the effects of tezosentan, a dual endothelin-receptor antagonist, on the cardiovascular and renal systems of neonatal piglets during endotoxemia. METHODS: Thirteen piglets were subjected to endotoxic shock and divided into a fluid-therapy group that received 0.9% normal saline and a group that received tezosentan (1 mg/kg/h). Mean arterial pressure (MAP), heart rate (HR), and glomerular filtration rate (GFR) were plotted at baseline, 1, 2, and 3 hours. Cardiac index (CI), renal blood flow (RBF), systemic vascular resistance (SVR), and renal vascular resistance (RVR) were obtained at baseline, 1, and 3 hours after baseline. RESULTS: (P <.05 for 3 hours versus baseline and tezosentan versus fluid). Although fluid therapy in endotoxemia had no significant effect on MAP and RVR, it significantly increased HR (139 plus minus 17 to 246 plus minus 17 beats/min) and SVR (0.08 plus minus 0.05 to 0.33 plus minus 0.09 mm Hg/mL/min) and decreased CI (407 plus minus 208 to 98 plus minus 13 mL/min/kg), RBF (1.84 plus minus 0.38 to 0.97 plus minus 0.34 mL/min/kg kidney), and GFR (0.20 plus minus 0.05 to 0.11 plus minus 0.04 mL/min/kg) at 3 hours. The use of tezosentan also significantly increased HR (130 plus minus 14 to 220 plus minus 31 beats/min), but unlike in the fluid therapy group, there was a significant fall in MAP (77 plus minus 10 to 54 plus minus 9 mm Hg) and RVR (1.92 plus minus 0.44 to 1.77 plus minus 0.64 mm Hg/mL/min) and a less severe decrease in CI (482 plus minus 188 to 176 plus minus 67 mL/min/kg) at 3 hours. SVR, RBF, and GFR were maintained. CONCLUSIONS: Endotoxic shock affected cardiac and renal functions in both treatment groups. Fluid therapy alone could not prevent a statistically significant fall in CI, RBF, and GFR or prevent the increase in HR and SVR. Endothelin antagonism with tezosentan resulted in a statistically significant fall in MAP and RVR from baseline, not seen in the fluid-therapy group. CI and RBF were significantly higher, and MAP, SVR, and RVR were significantly lower when compared with the fluid-therapy group at 3 hours. GFR also was maintained at baseline with tezosentan. During endotoxemia, endothelin antagonism maintained renal and cardiac functions better than with fluid therapy alone. PMID- 11877673 TI - In utero meconium exposure increases spinal cord necrosis in a rat model of myelomeningocele. AB - BACKGROUND/PURPOSE: The rationale for in utero repair of myelomeningocele has been supported experimentally by the observation of preserved neural function after prenatal closure of surgically created defects compared with nonrepaired controls. The mechanism of injury to the exposed neural elements is unknown. Postulated mechanisms include trauma to the herniated neural elements or progressive injury from amniotic fluid exposure as gestation proceeds. A component of amniotic fluid that may contribute to neural injury is meconium. In the current study the effect of human meconium on the exposed spinal cord in a fetal rat model of myelomeningocele was examined. METHODS: Twenty time-dated pregnant rats underwent laparotomy at 181/2 days of gestation. The exposed uterus was bathed in ritrodrine for tocolysis. The amniotic cavity was opened over the dorsal midline of the fetal rat, and, under a dissecting microscope (x25), a 2- to 3-level laminectomy was performed. Under magnification (x40), the translucent dura was opened using a 25-gauge needle as a knife. Two fetuses per dam were operated on. In the control group, the amniotic fluid was restored with saline solution, whereas in the experimental group a solution of Human meconium diluted (10%) in saline was used to restore the amniotic fluid. Fetuses were harvested by cesarean section at 211/2 days' gestational age. The liveborn pups were then killed and fixed in 10% formaline. Sections 10 micrometer thick were stained with H&E and studied by light microscopy for evidence of spinal cord injury. RESULTS: Seven of 20 (35%) experimental rat pups and 6 of 20 (30%) control rat pups were liveborn. All liveborn pups had severe paralysis of the hindlimbs and tail, so that functional differences between the 2 groups could not be detected. Histologic examination of 13 spinal cords at the site of surgical exposure showed that necrosis of neural tissue in 5 of 7 meconium-exposed rat pups was increased when compared with that observed in the 6 fetuses exposed to amniotic fluid without meconium. In general, inflammation was greater and repair processes appeared delayed in meconium-exposed rat pups. CONCLUSIONS: Exposure of the spinal cord of fetal rats to amniotic fluid by surgically created myelomeningocele leads to severe functional impairment. Histologically recognizable necrosis of neural elements was increased in those animals that were exposed to diluted human meconium in the amniotic fluid. The results support the hypothesis that meconium may contribute to the pathophysiology of spinal cord injury observed in myelomeningocele. PMID- 11877674 TI - Ad libitum feeding decreases hospital stay for neonates after pyloromyotomy. AB - PURPOSE: The optimal feeding regimen for neonates after pyloromyotomy for hypertrophic pyloric stenosis (HPS) remains controversial. This study sought to compare ad libitum feeding to a Conventional feeding regimen with regard to time to full diet, length of hospital stay, and readmission rates. METHODS: A 6-month review of 36 consecutive patients who underwent pyloromyotomy for HPS was undertaken. Patients were fed in 1 of 2 ways according to specific surgeon preference. Conventional Regimen patients (n = 19) were kept nothing by mouth (NPO) for 6 hours after surgery and incrementally advanced to full feedings. ad libitum (n = 17) patients were kept NPO until fully reversed from anesthesia and then given full strength formula or breast milk. Discharge was considered when 2 feedings of 60 mL were tolerated. RESULTS: Twenty-eight males and 8 females with a mean age of 5.0 plus minus 1.7 (SD) weeks, gestational age of 39 plus minus 2.1 weeks, weight of 4.0 plus minus 0.9 kg, and operating time of 56 plus minus 12 minutes were studied. The interval from operating room to full diet was significantly less with ad libitum feeding than on the conventional regimen (20.3 plus minus 5.0 v. 25.4 plus minus 8.3 hours, P <.05). The Ad Libitum group also had a significantly decreased length of hospital stay (28.5 plus minus 8.9 hours v. 35.8 plus minus 11 hours; P <.05). There were no readmissions in either group. CONCLUSIONS: Ad libitum feedings decrease time to full diet and discharge without an increase in readmission rates. The estimated potential savings per patient using ad libitum feedings were $392.00. Thus, the use of ad libitum feedings after pyloromyotomy for HPS appears indicated. PMID- 11877675 TI - The changing face of surgical indications for necrotizing enterocolitis. AB - PURPOSE: The aim of this study was to compare the proportion of operations for acute necrotizing enterocolitis (NEC) and post NEC strictures. METHODS: The authors reviewed 195 charts of children referred to our institution for NEC or post-NEC strictures between 1990 and 1999. Seventy-one children were classified as Bell stage I and were excluded. The remaining 124 patients were classified as either Bell stage II or III and formed the basis of our study. These patients were subdivided into 2 groups: (1) group I (n = 69) comprised patients treated from 1990 until 1994 and (2) group II (n = 55) from 1995 until 1999. Statistical analysis consisted of X(2) and Student's t tests. Significance occurred when P less-than-or-equal 0.05. RESULTS: Both groups were similar with regard to sex, obstetrical history, indomethacin use, umbilical artery catheter use, and enteral feeding. The total operative rate for all patients with either acute NEC or post NEC strictures increased over time from 46% (32 of 69) in group I to 69% (38 of 55) in group II (P <.01). Specifically, post-NEC stricture was the initial operation in 16% (5 of 32) of group I patients versus 37% (14 of 38) of group II patients (P <.05). Subdividing each group by method of treatment of their NEC showed that medically treated patients had an increased incidence of stricture over time (group I, 15% v. group II, 48%; P <.01). Surgically treated children maintained a similar rate of stricture (group I, 36% v. group II, 33%). The mortality rate was comparable in both groups. CONCLUSIONS: At our institution, the total operative rate for necrotizing enterocolitis has increased over the last 10 years. This is because of 2 factors: (1) an increase in the percentage of stage III patients and (2) an increase in referrals for post--necrotizing enterocolitis strictures. No specific criteria could be identified to predict which patients were at risk for post--necrotizing enterocolitis strictures after medical treatment. PMID- 11877676 TI - The role of protective antireflux procedures in neurologically impaired children: a decision analysis. AB - BACKGROUND/PURPOSE: Despite normal clinical history and preoperative radiologic and pH studies, gastroesophageal reflux (GER) can become apparent in neurologically impaired (NI) children after gastrostomy tube placement. An antireflux procedure performed at the time of gastrostomy tube placement may prevent postoperative GER and help avoid the need for a subsequent surgical procedure but is associated with a high morbidity and mortality rate in NI children. The purpose of this study was to determine the role of protective antireflux procedures in NI children undergoing gastrostomy tube placement. METHODS: Decision analysis was used to evaluate the effect of a protective antireflux procedure on morbidity and mortality in NI children. The rate of postoperative GER, need for secondary antireflux procedures, and morbidity and mortality rates after gastrostomy tube placement with or without an antireflux procedure in NI children were estimated from the literature and expert opinion and used to construct decision trees. RESULTS: At baseline values, gastrostomy tube placement resulted in a lower morbidity (11% v 13%) than gastrostomy tube placement with a protective antireflux procedure. One-way sensitivity analysis showed that gastrostomy tube placement was the favored approach when the morbidity of gastrostomy tube placement was less than 11% or the morbidity of antireflux surgery was greater than 10%. At baseline values, gastrostomy tube placement resulted in a lower mortality rate (0.3% v 0.8%) than gastrostomy tube placement with a protective antireflux procedure. Using 1-way sensitivity analysis, no threshold value of any variable was found that favored the use of a protective antireflux procedure with respect to mortality. CONCLUSIONS: Although a protective antireflux procedure may reduce the need for additional surgery, inclusion of this procedure is associated with a higher morbidity and mortality rate. Initial placement of a gastrostomy tube without a protective antireflux procedure is the favored approach for NI children without preoperative evidence of GER. PMID- 11877677 TI - The survivin:Fas ratio is predictive of recurrent disease in neuroblastoma. AB - BACKGROUND/PURPOSE: Several clinical and biologic features of neuroblastoma (NB) are used to predict the risk of recurrent disease. The balance between antiapoptotic and proapoptotic factors within a tumor may affect its ability to survive. Survivin is an antiapoptotic factor expressed in highly proliferative NB, whereas Fas is a proapoptotic factor that portends a favorable prognosis. The authors determined whether the ratio of survivin to Fas (S:F ratio) is predictive of recurrent disease in patients with NB. The authors previously have shown the S:F ratio is predictive of recurrent disease in pediatric renal tumors. METHODS: The authors quantified the levels of 9 different apoptotic mRNA species using Rnase Protection assay (RPA, Riboquant, PharMingen, San Diego, CA). Twenty-eight primary tumor specimens were evaluated from patients with ganglioneuroma (n = 3), ganglioneuroblastoma (n = 2), and neuroblastoma (n = 23) from tumors of all clinical stages obtained at the time of diagnosis. mRNA levels were calculated as a percentage of L32 for each specimen assayed, and positive expression was assumed to be greater than 10% of L32. RESULTS: Survivin was expressed in 90% of tumors that went on to recur and only in 27.7% of those that were cured. The S:F ratio was significantly greater in tumors that went on to recur (n = 10) compared with those from patients that were cured (n = 18) (median S:F ratio, 3.3 v 0.75; P =.0002, Wilcoxon rank-sum test). A cutoff ratio of 2.3 was highly predictive of tumor recurrence irrespective of clinical stage of disease (area under ROC curve = 0.906). Sensitivity was 80% (CI, 44.4% to 97.5%), specificity was 94.4% (CI, 72.7% to 99.9%), positive predictive value was 88.9% (CI, 51.8% to 99.7%), and negative predictive value was 89.5% (66.9% to 98.7%). Twenty-five of 28 (89.3%) tumor ratios were correct in predicting outcome. CONCLUSIONS: The survivin:Fas ratio in primary tumors may be used to predict the risk for recurrent disease in patients with NB. The S:F ratio appears to be a more sensitive predictor of recurrent disease than survivin expression alone. Determining this ratio may not only be helpful in guiding follow-up of patients with NB, but also may aid in stratifying patients for more aggressive therapeutic strategies. PMID- 11877679 TI - Cardiac relocation and chest wall reconstruction after separation of thoracopagus conjoined twins with a single heart. AB - Separation of thoracopagus conjoined twins with a single heart and the twin reversed arterial perfusion sequence yielded a single surviving infant with a protuberant heart covered by ribs and soft tissue from the nonsurviving twin. At 13 months of age, the heart was relocated in the chest after caudal mobilization of the diaphragms. The protective tissue cage was removed and a normal chest contour established. This technique also may be useful in the treatment of thoracic ectopia cordis. PMID- 11877678 TI - Pediatric vacuum packing wound closure for damage-control laparotomy. AB - BACKGROUND/PURPOSE: Recent studies advocate the use of the open-abdomen technique for managing the abdominal compartment syndrome and uncontrolled intraperitoneal sepsis. The authors have used vacuum packing (Vac-Pac) in pediatric patients with excellent results and have developed a method for closing widely distracted fascial edges to avoid the need for skin grafting. METHODS: Patients who had an intraabdominal catastrophe best managed by a temporary open-abdomen technique were included. After damage control laparotomy, the Vac-Pac temporary closure was used. In 2 patients a corsetlike lacing was used to bring the widely separated fascial edges together gradually. RESULTS: Five patients with intraabdominal sepsis and one with the abdominal compartment syndrome were included. The length of time the Vac-Pac was used ranged from 3 to 21 days. In 2 patients, the corset closure allowed wound approximation within 5 to 7 days. One patient died of overwhelming sepsis, the remainder of the patients survived. CONCLUSIONS: The Vac Pac technique for abdominal closure is a simple and inexpensive means by which to manage the open abdomen in the pediatric patient effectively. Use of a corset type closure for wounds with widely distracted edges should eliminate the need for skin grafting over an open abdomen. PMID- 11877680 TI - Distinct response of experimental neuroblastoma to combination antiangiogenic strategies. AB - BACKGROUND: The authors have shown previously that experimental neuroblastoma is partially inhibited (48%) by antivascular endothelial growth factor (anti-VEGF) antibody. The topoisomerase-I inhibitor, topotecan, has been shown to have antiangiogenic activity when administered in a low-dose, high-frequency regimen. We hypothesized that combining topotecan with anti-VEGF would suppress neuroblastoma more effectively than either agent alone. METHODS: A total of 10(6) neuroblastoma cells were implanted intrarenally in athymic mice. Animals received vehicle, topotecan, anti-VEGF, or topotecan plus anti-VEGF (n = 9, 20, 20, 20, respectively). All control and half the treated mice were killed at 6 weeks. Remaining (rebound) mice were maintained without treatment for 3 more weeks. Patterns of vasculature and apoptosis were determined immunohistochemically. RESULTS: Tumor weights at 6 weeks were reduced significantly in topotecan-only (0.07g) and combination-treated animals (0.08 g), compared with controls or anti VEGF--treated mice (1.18 g, 0.53 g; P <.0007, all). At 9 weeks, rebound tumor weights were greatest in anti-VEGF (2.82 g), intermediate in topotecan (1.82 g), and least in combination-treated animals (1.47 g); however, the only significant difference was between anti-VEGF and combination therapy (P = 0.04). All treated tumors were vascularized sparsely in comparison with controls at 6 weeks, but exhibited brisk neoangiogenesis at 9 weeks. CONCLUSIONS: Topotecan either with or without anti-VEGF antibody significantly suppresses neuroblastoma xenograft growth in comparison with controls or anti-VEGF antibody alone. Combining topotecan with anti-VEGF antibody significantly inhibited rebound tumor growth in comparison with anti-VEGF antibody alone. Combination therapy may improve durability of antiangiogenic inhibition of neuroblastoma. PMID- 11877681 TI - p53 accumulation in favorable-histology Wilms tumor is associated with angiogenesis and clinically aggressive disease. AB - BACKGROUND/PURPOSE: Unfavorable histology (UH) in Wilms tumor has been linked to malfunction of the p53 tumor suppressor gene, which regulates (1) the endogenous angiogenesis suppressor thrombospondin-1 (TSP-1) and (2) vascular endothelial growth factor (VEGF). The authors hypothesized that clinically aggressive favorable histology Wilms tumor (FH), like UH, but distinct from standard-risk FH disease, would display altered p53/TSP-1 function and upregulated angiogenesis. METHODS: Three Wilms tumor specimens manifesting different histology and clinical behavior were obtained: clinically aggressive UH, clinically aggressive FH, and standard-risk FH disease. Xenografts were induced intrarenally in athymic mice. P53, TSP-1, and VEGF status and neovascularity were assessed in tumor tissues. Lungs were evaluated for metastasis. RESULTS: Clinically aggressive FH Wilms tumor displayed progressive alteration in p53/TSP-1 status and upregulation of VEGF. Such alteration was observed in the UH tumor, but was absent from the standard-risk FH tumor. Xenografts from clinically aggressive tumors displayed brisk neoangiogenesis and yielded lung metastases. CONCLUSIONS: This is the first report of altered p53/TSP-1 function in association with clinically aggressive behavior in FH Wilms tumor. These characteristics were not observed in parallel studies of a nonaggressive FH tumor. Loss of wild-type p53 function may contribute to disease progression in FH Wilms tumor, in part by upregulation of VEGF. PMID- 11877682 TI - Resistance of a VEGF-producing tumor to anti-VEGF antibody: unimpeded growth of human rhabdoid tumor xenografts. AB - BACKGROUND/PURPOSE: Rhabdoid tumor of the kidney (RTK) is a lethal malignancy of childhood for which there currently are no effective therapies. Because vascular endothelial growth factor (VEGF) is nearly ubiquitous in human tumors, the authors hypothesized that a xenograft model of RTK would (1) express VEGF and (2) respond to anti-VEGF intervention. METHODS: A total of 2 x 10(6) cultured RTK cells were implanted intrarenally (G-401) in athymic mice. Control/treated animals received either vehicle (phosphate-buffered saline, PBS) or anti-VEGF antibody (anti-VEGF) for 5 weeks (n = 20, 17, respectively). Vasculature was mapped by angiography and immunostaining. Apoptosis was assessed by TdT-mediated dUTP nick end labeling (TUNEL) assay, VEGF expression examined by reverse transcription polymerase chain reaction, and tumor weights compared by Kruskal Wallis analysis. RESULTS: Mean tumor weights were not altered significantly by anti-VEGF (0.78-g, controls v 0.56-g treated tumors; P value, not significant). Grossly, xenografts grew in a novel manner, encasing rather than invading the kidney, and did not metastasize. PECAM-1 immunostaining and fluorescein angiography showed similar vascularity in control and treated xenografts. Both apoptosis and VEGF expression were unchanged in treated specimens. CONCLUSIONS: Unexpectedly, growth of RTK xenografts was not inhibited by specific anti-VEGF antibody, although these tumors express significant amounts of VEGF. In addition, RTK vasculature, apoptosis, and VEGF expression were not substantially altered by anti-VEGF antibody. These results suggest that tumor-derived VEGF is of highly variable importance in different malignancies. PMID- 11877683 TI - Pyriform sinus malformations: a cadaveric representation. AB - BACKGROUND/PURPOSE: The most important aspects in management of pyriform sinus malformations are awareness of the diagnosis, familiarity with the clinical manifestations, and complete surgical excision of the entire tract. Pyriform sinus anomalies are the least common branchial apparatus malformations and present anatomically as sinus tracts with or without cystic dilatation. The clinical presentations can include lateral neck mass, thyroid abscess, suppurative thyroiditis, retropharyngeal abscess, neonatal airway obstruction, and even carcinoma. Recurrent symptoms after surgery suggest incomplete identification and excision of the tract. METHODS: Cadaveric dissections were performed to show both the proposed embryologic course and clinical manifestations of third and fourth branchial apparatus pyriform sinus anomalies. RESULTS: Illustrations and digital camera images of the cadaveric models are presented to explain the course of pyriform sinus fistula tracts. CONCLUSIONS: The authors discuss 3 case presentations of pyriform sinus anomalies with emphasis on their proposed embryologic origin and anatomic basis for surgical management. Surgical excision is the mainstay of therapy. Understanding the embryologic basis for pyriform sinus malformations aids in recognition of the diagnosis despite the myriad of clinical presentations. Laryngoscopy with sinus cannulation facilitates removal of the entire sinus tract with preservation of the recurrent and superior laryngeal nerves. PMID- 11877685 TI - Laparoscopic approach to surgical management of congenital diaphragmatic hernia in the newborn. PMID- 11877684 TI - Effective targeted cytotoxicity of neuroblastoma cells. AB - BACKGROUND/PURPOSE: Despite aggressive treatment with surgery, chemotherapy, and radiotherapy, the prognosis for many children with neuroblastoma remains poor. Targeted toxins represent novel cancer therapeutics designed to selectively target and kill cancer cells. The authors have developed a novel fusion toxin, DT5F11, consisting of truncated diphtheria toxin (DT(A)) linked to a single chain antibody (sc5F11) targeting the GD(2) antigen found on most neuroblastoma cells. This report describes the construction, expression, and in vitro function of DT5F11. METHODS: Utilizing restriction enzyme digestion, polymerase chain reaction amplification, and gel electrophoresis, the prkDTL5F11 plasmid was created by the fusion of distinct coding sequences for a single-chain GD(2) targeting antibody (sc5F11) and truncated diphtheria toxin (DT(A)). DH5alpha Escherichi coli-competent cells were transformed with prkDTL5F11; DNA was amplified, isolated, and sequenced. The fusion protein was expressed and assayed by Western blot. Targeted cytotoxicity was analyzed on GD(2)-positive (SK-N-AS, IMR-32, SK-N-MC, LAN-1) and GD(2)-negative (HeLa) cells. RESULTS: Fluorescent dye labeled cycle sequencing identified the constructed fusion toxin gene. Western blot analysis using a mouse antihuman DT(A) antibody showed a 69-kD band identifying the fusion toxin, DT5F11. Targeted cell killing with DT5F11 was seen only in GD(2) positive cells. CONCLUSIONS: This study demonstrates creation of a novel fusion toxin with effective GD(2)-targeted cellular toxicity. Further investigation of this fusion toxin as a therapeutic agent in the management of neuroblastoma is warranted. PMID- 11877686 TI - Benefits, morbidity, and mortality associated with long-term administration of oral anticoagulant therapy to patients with peripheral arterial bypass procedures: a prospective randomized study. AB - BACKGROUND: The benefits of the long-term administration of oral anticoagulant therapy remain unclear in patients with lower extremity arterial bypass surgery. We studied the effect of warfarin plus aspirin therapy (WASA) versus aspirin therapy alone (ASA) on patient mortality, morbidity and bypass patency rates in a randomized clinical trial. METHODS: In a multicenter, prospective, nonmasked clinical trial, 831 patients who underwent peripheral arterial bypass surgery were compared in a long-term treatment program of WASA (target international normalized ratio of 1.4 to 2.8; 325 mg/day) with ASA (325 mg/day). The primary end point was bypass patency, and mortality and morbidity were the secondary endpoints. RESULTS: There were 133 deaths in the WASA group (31.8%) and 95 deaths in the ASA group (23.0%; risk ratio, 1.41; 95% confidence interval, 1.09 to 1.84; P =.0001). Major hemorrhagic events occurred more frequently in the WASA group (WASA, n = 35; ASA, n = 15; P =.02). In the prosthetic bypass group, there was no significant difference in patency rate in the 8-mm bypass subgroup, but there was a significant difference in patency rate in the 6-mm bypass subgroup (femoral popliteal; 71.4% in the WASA group versus 57.9% in the ASA group; P =.02). In the vein bypass group, patency rate was unaffected (75.3% in the WASA group versus 74.9% in the ASA group). CONCLUSION: The long-term administration of warfarin therapy when combined with aspirin therapy has only a few selected indications for improvement of bypass patency and is associated with an increased risk of morbidity and mortality. PMID- 11877687 TI - Subclavian carotid transposition and bypass grafting: consecutive cohort study and systematic review. AB - PURPOSE: We describe outcomes in a cohort of patients undergoing subclavian carotid transposition (SCT) for occlusive disease of the first segment of the subclavian artery and perform a systematic review of the literature on SCT and carotid subclavian bypass grafting (CSB). METHODS: Relevance, validity and extraction of review results were done in duplicate. Data were collected prospectively in our consecutive cohort of patients. RESULTS: From September 1990 to February 2001, we performed 27 SCTs, four for aneurysmal disease and 23 for occlusive disease. SCTs done for aneurysms were excluded from the current analysis. In patients with occlusive disease, the primary indications for surgery were vertebrobasilar and carotid symptoms (10, 44%), vertebrobasilar insufficiency (7, 30%), vertebrobasilar and arm symptoms (4, 17%), carotid symptoms (1, 4%), and vertebrobasilar, carotid, and arm symptoms (1, 4%). An SCT was performed in conjunction with an endarterectomy of the carotid artery in 12 patients (52%), with an endarterectomy of the subclavian artery in seven patients (30%), and with an endarterectomy of the vertebral artery in six patients (26%). A lymph leak complicated two surgeries (9%). In our series, patients improved clinically after surgery, and reconstructions were all found to be patent by means of Doppler ultrasound scanning at a mean follow-up of 25 +/- 21 months. Three patients (13%) died during follow-up of complications of coronary artery disease. From 1966 to 2000, 516 patients who underwent CSB and 511 patients who underwent a SCT were reported in the literature. Patency rates were 84% and 98%, respectively (P <.0001; absolute risk reduction, 15%; number-needed-to-treat differently, 7), and the rates of freedom from symptoms were 88% and 99%, respectively, at a mean follow-up of 59 +/- 17 months (range, 1-228 months). CONCLUSION: Our cohort study showed that SCT is safe and effective for reconstruction of the first segment of the subclavian artery. The systematic review suggested that rates of patency and freedom from clinical symptoms are higher with SCT than with CSB. PMID- 11877689 TI - Carotid artery closure for endarterectomy does not influence results of angioplasty-stenting for restenosis. AB - OBJECTIVE: Carotid angioplasty and stenting (CAS) has been recommended by some authors for the management of postendarterectomy restenosis. However, some authors have expressed concern about the influence of primary closure and patch angioplasty performed during carotid endarterectomy (CEA) on the incidence rate of complications after CAS. METHODS: We analyzed our consecutive series of 54 CAS procedures performed for restenosis after prior CEA. These procedures accounted for 75% of the 72 CAS procedures performed at our institution for all indications during the last 4 years. Of these 54 patients, 28 (52%) were men and 26 (48%) were women, with a mean age of 69 years. The mean clinical follow-up period was 18 months (range, 1 to 48 months). The mean interval between prior CEA and CAS was 16 months (range, 6 to 62 months). Nineteen patients were symptomatic (35%), and 35 were asymptomatic (65%). The mean severity of restenosis was 84% +/- 7% (standard deviation). The mean residual stenosis after CAS was 8% +/- 3% (standard deviation). RESULTS: Among the 54 prior CEAs, eight cases were performed with primary closure (15%), five procedures used patch closure with autologous vein (9%), and 41 operations used Dacron patch closures (76%). All patients were managed successfully with CAS with predeployment angioplasty with low profile balloons, self-expanding stents, and poststent angioplasty to approximate the transverse diameter of the carotid artery. No instances of contrast extravasation, arterial disruption, or subintimal dissection were observed. One stroke (1.8%), a retinal infarction with partial field of vision loss, occurred in a patient with prior CEA and Dacron patch closure, and no deaths were observed in the series. CONCLUSION: Performance of CAS for restenosis after CEA with autologous or synthetic patch angioplasty was technically successful in all 54 procedures. The method of closure of the arteriotomy during CEA, primary closure or patch angioplasty, did not influence the incidence of complications. PMID- 11877688 TI - Carotid artery stenting in a vascular surgery practice. AB - PURPOSE: We tested the clinical applicability, technical results, and morbidity of carotid angioplasty-stenting (CAS) in the treatment of severe stenosis of the internal carotid artery (ICA) in patients deemed to be high-risk candidates for carotid endarterectomy (CEA). METHOD: After an initial series (1994-1997) of 52 interventions, we adopted the use of a transfemoral access technique and self expanding stents in late 1997. From Dec 1, 1997, to Mar 31, 2001, 135 CAS procedures were performed on 132 patients with more than 70% (symptomatic) or more than 80% (asymptomatic) stenoses of the ICA. Sixty percent of the patients had no symptoms, and 40% of patients had symptoms. The interventional technique was standardized with the use of a 7F long interventional sheath, balloon pre dilatation of the stenotic lesion, placement of a self-expanding stent (Wallstent in 12 patients and a SMART stent in 120 patients), and post-balloon dilatation when necessary. Brain protection devices were not used. Patients were given clopidogrel and aspirin before and after the procedure and heparin during the intervention. RESULTS: All procedures except two were completed as planned, with access failure in three patients (2.2%). Residual in-stent stenosis of less than 20% was detected in 14 of 132 stented vessels (11%) and accepted as a satisfactory angiographic outcome. Neurologic complications included one patient with a single-episode transient ischemic attack (TIA; motor-sensory deficit of the hand) occurring 2 hours after CAS. One patient sustained a major stroke after thrombosis of the stented ICA, which occurred 3 days after the CAS procedure and 24 hours after open-heart surgery. A third patient sustained a minor stroke that began intraprocedurally after post-balloon dilatation of the stent, and a fourth patient had another minor stroke with transient aphasia (beginning during the procedure and resolving after 4 hours) and monoparesis of the hand, which resolved after 1 week. All stented vessels remained patent during the follow-up period (range, 2-41 months; mean, 16 plus minus 9 months), with four instances of hemodynamically significant in-stent restenosis. Re-intervention with balloon angioplasty was undertaken successfully at 4 months in one patient with restenosis. The periprocedural mortality rate was 0. CONCLUSION: Carotid stenting can be performed with acceptable safety on carefully selected patients by using meticulous, standardized interventional techniques. It may offer a possibly superior therapeutic alternative for non-CEA candidates. Evolving technological improvements and brain protection devices are likely to enhance its role in the treatment of carotid artery disease in the future. Surgical endarterectomy remains the standard of care for most patients at the present time. PMID- 11877690 TI - Internal carotid artery flow volume measurement and other intraoperative duplex scanning parameters as predictors of stroke after carotid endarterectomy. AB - PURPOSE: Intraoperative duplex scanning (IDS) after carotid endarterectomy (CEA) has been shown to reliably identify major defects either by significant changes in peak systolic velocities or by B-mode imaging. To evaluate whether IDS could also predict postoperative strokes in technically flawless CEAs, we analyzed several hemodynamic parameters and correlated them with patient outcome. METHODS: From March 2000 to February 2001, 226 consecutive primary CEAs were performed in 208 patients (120 men). Of these, 153 lesions were asymptomatic. General anesthesia and synthetic carotid artery patches were used routinely. Intraluminal shunts were used when internal carotid artery (ICA) back-pressures were <50 mm Hg (35% of cases). IDS consisted of B-mode and color-flow imaging and spectral analyses of the common, external, and internal carotid arteries. Volume flows were measured three times, and the mean flow rate was used for this study. RESULTS: The first set of data was analyzed when the twenty-ninth patient had the second immediate postoperative stroke. It was noted that the two patients who had postoperative strokes had mean ICA volume flows (MICAVF) of 48 mL/min and 85 mL/min. Only two additional patients had MICAVF <100 mL/min. The remaining 25 cases had MICAVF ranging from 102 to 299 mL/min, with a mean of 165 +/- 57 mL/min (+/-SD) (P <.02). Although there was a significant correlation between MICAVF and ICA peak systolic velocity (P <.01), the latter was not found to be a significant predictor of postoperative stroke. Moreover, end-diastolic velocities, resistive index, ICA diameter, and ICA back-pressure also did not correlate with neurologic events. These findings led us to change our protocol for patients with MICAVF <100 mL/min. This included a repeat set of volume flow measurements after 15 to 20 minutes, withholding the reversal of heparin, and the liberal use of completion arteriography. Of the following 197 CEAs, 26 (13%) were found to have MICAVF <100 mL/min (range 55 to 99 mL/min; mean 79 +/- 18 mL/min). Of these, five had arteriography that documented spasm of the intracranial portion of the ICA in four and a small-diameter ICA (<2 mm) in one. Except for the five cases, the remaining 21 cases had MICAVF >100 mL/min (range 105 to 158 mL/min, mean 127 +/- 20 mL/min [+/-SD]) on repeat study. Four patients with persistent ICA low flow (70 to 99 mL/min) were treated with postoperative anticoagulation. One of the last 197 patients had a stroke caused by hyperperfusion syndrome 2 weeks after operation. Overall, six of 226 cases (2.7%) required revision on the basis of abnormal B-mode imaging results or peak systolic velocities >150 cm/s. There were two common carotid artery flaps, two ICA stenoses, one ICA flap, and one localized thrombus. All six were successfully revised and had repeat normal IDS study results, and none of these patients had a postoperative stroke. CONCLUSIONS: IDS is helpful in identifying residual lesions or defects that may contribute to postoperative neurologic deficits. MICAVF <100 mL/min are suggestive of spasm that could lead to thrombus formation and stroke, particularly in the presence of synthetic patches. We suggest that heparin reversal should not be used unless ICA flow rates are >100 mL/min. ICA spasm is short lived in most patients undergoing CEA. PMID- 11877692 TI - Long-term outcome after mesenteric artery reconstruction: a 37-year experience. AB - PURPOSE: The purpose of this study was the definition of the late results and determining factors after mesenteric artery reconstruction (MAR) for atherosclerotic mesenteric ischemia. METHODS: A retrospective review identified 48 consecutive patients (66 arteries) who underwent MAR for acute mesenteric ischemia (AMI) of nonembolic origin (n = 23; 12 with and 11 without prior symptoms) and chronic mesenteric ischemia (CMI; n = 25) from 1963 to 2000 in a tertiary care referral center. The 29 women (60%) and the 19 men (40%) had a mean age of 64 years (range, 40 to 87 years). The operative procedures consisted of bypass grafting in 36 arteries (AMI, 12; CMI, 24), local endarterectomy (LEA) in 16 arteries (AMI, 9; CMI 7), and transaortic endarterectomy (TAE) in 14 arteries (AMI, 4; CMI, 10). The follow-up of the 34 survivors was complete in all but four patients and averaged 5.3 years (range, 30 days to 36 years). Radiographic documentation of vessel/graft patency was obtained in 33 of 34 survivors. RESULTS: Single-vessel revascularization was performed more frequently in the AMI group than in the CMI group (91% versus 48%; P =.001). The perioperative (<30 days) mortality rate in the AMI group was 52% (12 of 23 cases) as compared with 0 of 25 cases in the CMI group (P <.001). Bowel infarction was the cause of nine deaths. Major complications occurred in 60% of the cases. Fifteen late graft failures occurred, for a cumulative patency rate of 57% at 5 years and 46% at 10 years. TAE was associated with improved patency rates as compared with LEA (TAE versus LEA; P =.002). Symptomatic recurrences developed in eight patients, all involving superior mesenteric artery thrombosis (P <.001). The freedom-from recurrence rates in the survivors were 79% at 5 years and 59% at 10 years. The late survival rates were 54% and 20% at 5 and 10 years, respectively. With the exclusion of perioperative deaths, the probability of long-term survival was 77% at 5 years and 29% at 10 years and did not differ between AMI and CMI. CONCLUSION: Although MAR for CMI carries a low mortality rate, AMI remains a lethal and frequently unheralded problem. Long-term patency and symptom-free survival can be expected after successful MAR for AMI and is comparable with those rates achieved after MAR for CMI. The patency of the SMA is important in the prevention of symptomatic recurrences. Elective MAR is indicated in patients with CMI and warrants long-term surveillance. PMID- 11877691 TI - Contemporary management of acute mesenteric ischemia: Factors associated with survival. AB - PURPOSE: Acute mesenteric ischemia (AMI) is a morbid condition with a difficult diagnosis and a high rate of complications, which is associated with a high mortality rate. For the evaluation of the results of current management and the examination of factors associated with survival, we reviewed our experience. METHODS: The clinical data of all the patients who underwent operation for AMI between January 1, 1990, and December 31, 1999, were retrospectively reviewed, clinical outcome was recorded, and factors associated with survival rate were analyzed. RESULTS: Fifty-eight patients (22 men and 36 women; mean age, 67 years; age range, 35 to 96 years) underwent study. The cause of AMI was embolism in 16 patients (28%), thrombosis in 37 patients (64%), and nonocclusive mesenteric ischemia (NMI) in five patients (8.6%). Abdominal pain was the most frequent presenting symptom (95%). Twenty-five patients (43%) had previous symptoms of chronic mesenteric ischemia. All the patients underwent abdominal exploration, preceded with arteriography in 47 (81%) and with endovascular treatment in eight. Open mesenteric revascularization was performed in 43 patients (bypass grafting, n = 22; thromboembolectomy, n = 19; patch angioplasty, n = 11; endarterectomy, n = 5; reimplantation, n = 2). Thirty-one patients (53%) needed bowel resection at the first operation. Twenty-three patients underwent second-look procedures, 11 patients underwent bowel resections (repeat resection, n = 9), and three patients underwent exploration only. The 30-day mortality rate was 32%. The rate was 31% in patients with embolism, 32% in patients with thrombosis, and 80% in patients with NMI. Multiorgan failure (n = 18 patients) was the most frequent cause of death. The cumulative survival rates at 90 days, at 1 year, and at 3 years were 59%, 43%, and 32%, respectively, which was lower than the rate of a Midwestern white control population (P <.001). Six of the 16 late deaths (38%) occurred because of complications of mesenteric ischemia. Age less than 60 years (P <.003) and bowel resection (P =.03) were associated with improved survival rates. CONCLUSION: The contemporary management of AMI with revascularization with open surgical techniques, resection of nonviable bowel, and liberal use of second-look procedures results in the early survival of two thirds of the patients with embolism and thrombosis. Older patients, those who did not undergo bowel resection, and those with NMI have the highest mortality rates. The long-term survival rate remains dismal. Timely revascularization in patients who are symptomatic with chronic mesenteric ischemia should be considered to decrease the high mortality rate of AMI. PMID- 11877693 TI - Significance of endoleaks after endovascular repair of abdominal aortic aneurysms: The EUROSTAR experience. AB - OBJECTIVE: The purpose of this study was to assess the incidence, risk factors, and consequences of endoleaks after endovascular repair of abdominal aortic aneurysm. METHODS: Data on 2463 patients were collected from 87 European centers and recorded in a central database. Preoperative data were compared for patients with collateral retrograde perfusion (type II) endoleak (group A), patients with device-related (type I and III) endoleaks (group B), and patients in whom no endoleak was detected (group C). Only endoleaks observed after the first postoperative month of follow-up were taken into consideration. Regression analysis was performed to investigate statistical relationships between the occurrence and type of endoleak and preoperative patient and morphologic characteristics, operative details, type of device, and experience of the operating team. In addition, postoperative changes in aneurysmal morphology, the need for secondary interventions, conversions to open repair, aneurysmal rupture, and mortality during follow-up were compared between these study groups. RESULTS: Patients in group A had a higher prevalence of a patent inferior mesenteric artery compared with patients without endoleak. Patients in group B were treated more frequently than patients in group C by an operating team with experience of less than 30 procedures. The mean follow-up period was 15.4 months. Secondary interventions were needed in 13% of the patients. Rupture of the aneurysm during follow-up occurred in 0.52% (1/191) in group A, 3.37% (10/297) in group B, and 0.25% (5/1975) in group C. Life table analysis comparing the three study groups demonstrated a significantly higher rate of rupture in group B than in group C (P =.002). The incidence of conversion to open repair during follow-up was higher in group B than in the other two study groups (P <.01). Death was related to the aneurysm or to endovascular repair of the aneurysm in 7% of patients. Secondary outcome success, defined as absence of rupture and conversion, was significantly higher in group A and C compared with that in group B (P =.006 and P =.0001, respectively). CONCLUSIONS: The presence of device-related endoleaks correlated with a higher risk of aneurysmal rupture and conversion compared with patients without type I or III endoleaks. Type II endoleak was not associated more often with these events. Consequently, intervention in type II endoleak should only be performed in case of increase of aneurysm size. PMID- 11877694 TI - The incidence and natural history of type I and II endoleak: a 5-year follow-up assessment with color duplex ultrasound scan. AB - PURPOSE: The purpose of this study was the demonstration of the value of color duplex ultrasound (CDU) scanning in the detection of type I endoleak (T1EL) and type II endoleak (T2EL), the correlation of Doppler scan waveform pattern to endoleak persistence or seal, and the description of the natural history of endoleak. METHODS: The study was a retrospective review of 83 patients who underwent periodic CDU scan and computed tomographic (CT) scan surveillance of the endograft and aneurysm sac after insertion of an aortic endograft for abdominal aortic aneurysm (AAA). Forty-one patients (49%) with an endoleak at anytime in the follow-up period form the basis of this report. RESULTS: T1EL was detected in all five patients with CDU and CT scans. T2EL was detected in 36 patients with CDU scan as compared with 18 patients with CT scan. With CT scan, endoleak was not detected when CDU scan showed no endoleak. Conversely, all CT scan--detected endoleaks were found with CDU scanning. The T2EL source artery was identified with CT scan in seven patients, whereas the source was identified in all 36 patients with CDU scan. Endoleak source did not correlate with outcome (seal or persistence). However, a to/fro Doppler scan waveform pattern was associated with spontaneous T2EL seal in seven of 12 patients, and a monophasic or biphasic waveform was associated with endoleak persistence in 14 of 17 patients (P =.023, with chi(2) test). Thirteen of 36 T2ELs underwent spontaneous seal by 6.2 +/- 2.8 months. T2ELs without increasing AAA diameter were observed. Eight patients with persistent T2EL present for more than 12 months did not undergo treatment. However, two patients underwent T2EL obliteration with coils because of AAA sac enlargement. T1EL of the distal attachment site was the initial endoleak identified in five patients, but seven patients harboring T2ELs had subsequent T1ELs develop. For the entire 83 patients, the combined T1EL and T2EL prevalence rate was 20% of patients at a 6-month follow-up period, but this rate increased to 50% after 24 months. The incidence rate of newly detected endoleaks and of spontaneous sealing was 24.4% at 12 months and 12.5% in longer term follow-up period. CONCLUSION: CDU scan is effective in the identification of the type of endoleak, the delineation of the vessel involved, and the hemodynamic information not available with any other testing method. Endoleaks have a dynamic natural history characterized by a variable onset with changing branch vessel involvement and spectral flow patterns. Periodic long-term endograft surveillance with CDU scanning is necessary for following existing endoleaks and for detecting new ones. Corroboration of these findings in larger multicenter prospective trials will be needed to determine whether CDU scan analysis of endoleaks would be predictive of long-term success in endovascular AAA repair. PMID- 11877695 TI - Aortic neck angulation predicts adverse outcome with endovascular abdominal aortic aneurysm repair. AB - BACKGROUND: Significant aortic neck angulation may predispose to suboptimal outcome after endovascular abdominal aortic aneurysm (EAAA) repair. However, the definition of "significant" neck angulation and its correlation with adverse outcome are poorly characterized. METHODS: Prospectively collected data on 148 consecutive EAAA repairs performed between December 1995 and January 2001 were supplemented with retrospective review of charts and radiographs. Aortic neck angulation was measured from arteriograms or three-dimensional computed tomography scanning reconstructions. Patients were excluded (n = 24) if radiographs were unavailable for review. Because of a paucity of severe aortic neck angulation in other endograft groups, only patients treated with a modular bifurcated device (Medtronic) (n = 81) were included in the final analysis. Mean time from implantation was 26.6 +/- 9.2 months. RESULTS: The risk of a patient experiencing one or more adverse events was 70%, 54.5%, and 16.6% in those with severe (>or=60 degrees, n = 10), moderate (40 to 59 degrees, n = 11), and mild (<40 degrees, n = 60) aortic neck angulation, respectively (P =.0003). Adverse events included death within 30 days (20% vs 0%, P =.0007), acute conversion to open repair (20% vs 0%, P =.0007), aneurysm expansion (9.1% to 20% vs 1.7%, P =.034), device migration (20% to 30% vs 3.3%, P =.013), and type I endoleak (23.8% vs 8.3%, P =.033), all occurring with significantly greater incidence in patients with moderate or severe aortic neck angulation when compared with those with mild angulation, respectively. Aortic neck length and diameter, age, and medical comorbidities were not significantly different between groups. CONCLUSION: Aortic neck angulation appears to be an important determinant of outcome after EAAA repair. Although patients with mild angulation (<40 degrees) had favorable outcomes in this series, those with moderate (40 to 59 degrees) or severe angulation (>or=60 degrees) had a 54% to 70% risk of one or more adverse events. Importantly, these outcomes occurred in spite of an adequate length (>2 cm) of proximal aortic neck. On the basis of these data, great caution should be exercised in recommending EAAA repair for patients with aortic neck angulation >or=40 degrees. PMID- 11877696 TI - Success of thrombolysis as a predictor of outcome in acute thrombosis of popliteal aneurysms. AB - PURPOSE: Acute limb ischemia after thrombosis of a popliteal aneurysm is a distinct and limb-threatening entity. Preoperative intra-arterial thrombolysis may improve the outcome in this challenging situation. This study retrospectively analyzed a consecutive series of patients treated with preoperative thrombolysis and subsequent revascularization. METHODS: Thirteen patients with acute limb ischemia caused by thrombosis of a popliteal aneurysm underwent catheter-directed intra-arterial thrombolysis with urokinase and subsequent vascular reconstruction. The angiographic and clinical outcome was analyzed and compared with that in the literature. RESULTS: Complete aneurysm thrombosis with absence of runoff was documented in 12 cases. Thrombolysis restored perfusion with patency of the popliteal artery and a one- or two-vessel runoff in 77% of cases (10/13). Early cumulative graft patency and limb salvage rates were 68% and 83%, respectively, with an ankle/brachial index of 0.8 +/- 0.2. Lytic failure followed by attempts at bypass grafting was present in three patients (23%) and resulted in above-knee amputation. Severe rhabdomyolysis and fatal pulmonary embolism were responsible for a 15% early mortality rate. CONCLUSION: Preoperative thrombolysis followed by bypass grafting is a valid treatment option for patients who can withstand an additional period of ischemia that does not require immediate revascularization and intraoperative lysis. Lytic failure identifies patients with a highly compromised runoff who are probably best treated by means of subsequent amputation, without any attempts at bypass grafting. PMID- 11877697 TI - Angiographic scoring of vascular occlusive disease in the diabetic foot: relevance to bypass graft patency and limb salvage. AB - OBJECTIVE: We graded the severity of occlusive disease in foot vessels of patients with diabetes and correlated the scoring obtained with graft patency and limb salvage. METHODS: In this retrospective review of 199 limbs studied by means of angiography in 117 patients with diabetes mellitus, 124 limbs underwent bypass grafting. Each dorsalis pedis (DP), lateral plantar (LP), and medial plantar (MP) artery was assigned a score according to the reporting standards of the Joint Vascular Societies Council (0, no stenosis > 20%; 1, 21%-49% stenosis; 2, 50%-99% stenosis; 2.5, < half the vessel length occluded; 3, > half the vessel length occluded.) A foot score (DP + MP + LP + 1) was calculated for each foot (1 to 10). The mean follow-up period was 14 months. RESULTS: For all 199 limbs and for the 124 limbs that underwent bypass grafting, the mean scores were similar for the DP, MP, and LP (1.8 +/- 1.0, 1.9 +/- 1.0, 1.9 +/- 1.0, respectively; P >.4). Only the MP and LP correlated with each other (r = 0.57; P <.0001). There were no scoring differences between limbs with symptoms and limbs that did not undergo bypass grafting. Bypass graft patency correlated with both the foot score and the MP score for tibial and inframalleolar grafts (P <.04). Patency correlated with the LP score only for inframalleolar bypass grafting procedures. The DP score alone did not differ between bypass grafts that remained patent and bypass grafts that failed. Bypass grafts in limbs with a foot score less than 7 and an MP score less than 2 had only a 2% failure rate. A foot score greater or equal to 7 was associated with a 30% failure rate for all bypass grafts (41% for inframalleolar grafts). Bypass grafts with low foot scores that failed did so much later than bypass grafts with high foot scores that failed (17 +/- 11 months vs 6 +/- 8 months; P <.02), possibly reflecting different etiologies for the failure. The limb salvage rate correlated with foot score (P <.05). The limbs that were saved had an average foot score of 6.4 +/- 2.0, versus 7.2 +/- 1.4 for limbs that required amputation. CONCLUSION: In patients with diabetes mellitus, the foot score is a useful tool for predicting the likelihood of graft patency and limb salvage for infrapopliteal revascularization. However, the relatively high bypass success rate (70%) in the presence of a high foot score (>or= 7) does not allow its use in identifying the subgroup of patients who are unlikely to benefit from bypass grafting surgery. It cannot be used as a means of selecting patients for primary amputation. PMID- 11877698 TI - Cutaneous microcirculation in the neuropathic diabetic foot improves significantly but not completely after successful lower extremity revascularization. AB - OBJECTIVE: The purpose of this study was the examination of the effect of successful large vessel revascularization on the microcirculation of the neuroischemic diabetic foot. RESEARCH DESIGN AND METHODS: We measured the cutaneous microvascular reactivity in the foot in 13 patients with diabetes with peripheral arterial disease and neuropathy (group DI) before and 4 to 6 weeks after successful lower extremity arterial revascularization. We also compared them with age-matched and sex-matched groups of 15 patients with diabetes and neuropathy, seven patients without neuropathy, and 12 healthy patients for control. We used single-point and laser Doppler scan imaging for the measurement of the foot skin vasodilatation in response to heating to 44 degrees C and to iontophoresis of 1% acetylcholine (endothelial-dependent response) and 1% sodium nitroprusside (endothelial-independent response). RESULTS: The group DI response to heat increased from 289% +/- 90% before surgery (percent increase over baseline measured in volts) to 427% +/- 61% (P <.05) after surgery but was still comparable with the response of the patients with diabetes and neuropathy (318% +/- 51%) and lower than the responses of the patients without neuropathy (766% +/ 220%) and the healthy patients for control (891% +/- 121%; P <.0001). The group DI acetylcholine response also improved from 6% +/- 4% before surgery to 26% +/- 8% after surgery (P <.05) and was similar to the responses of patients with diabetes and neuropathy (18% +/- 3%) and patients without neuropathy (38% +/- 8%) but still lower when compared with the response of the patients for control (48% +/- 9%; P <.001). The sodium nitroprusside response for group DI improved from 10% +/- 4% to 29% +/- 9% (P <.05) and was similar to the responses of the neuropathic (25% +/- 9%), nonneuropathic (32% plus minus 6%), and control (40% +/ 5%) groups. The group DI neurovascular response, which depends on the healthy function of the C-fiber nociceptors, was similar at baseline (5% +/- 9%) and after surgery (14% +/- %10) and in the neuropathic group (33% +/- 21%), but it was dramatically reduced when compared with the nonneuropathic (110% +/- 40%) and control (198% +/- 54%) groups (P <.001). CONCLUSION: Impaired vasodilation in the diabetic neuropathic lower extremity leads to functional ischemia, which improves considerably but is not completely corrected with successful bypass grafting surgery. Therefore, patients with diabetes and neuropathy may still be at high risk for the development of foot ulceration or the failure to have an existing ulcer heal despite adequate correction of large vessel blood flow. PMID- 11877699 TI - Age at onset of smoking is an independent risk factor in peripheral artery disease development. AB - PURPOSE: The potential effects of age at onset of smoking on cardiovascular diseases have been studied little, in contrast to the well-established evidence supporting a causal role of cigarette smoking in these diseases. We sought to analyze the relationship between age at smoking onset and development of symptomatic peripheral arterial occlusive disease (PAOD). METHODS: A population based sample of 573 active or former male smokers aged 55 to 74 years were studied. Present or previous symptomatic PAOD was confirmed by noninvasive testing. RESULTS: Sixty-one subjects (10.6%) had symptomatic PAOD. Prevalence of disease increased with earlier starting age (15.6% if 16 years) of smoking. After controlling for risk factors that meet confounding factor criteria (ie, subject age and number of pack-years), men who started smoking at age 16 or earlier had a substantially higher risk for development of PAOD (odds ratio, 2.19; 95% CI, 1.15-4.15; P =.016) than men who began to smoke at a later age. CONCLUSIONS: A starting age for smoking of 16 years or earlier more than doubles the risk of future symptomatic PAOD regardless of the amount of exposure to cigarette smoking. PMID- 11877700 TI - Sex-related differences in outcome after vascular interventions for lower limb ischemia. AB - OBJECTIVE: The purpose of this study was to investigate sex-related differences in outcome, defined as amputation rate and survival in patients treated for lower limb ischemia. MATERIAL AND METHOD: Age, sex, and type of procedure for all patients undergoing vascular interventions for lower limb ischemia in Stockholm, Sweden, from 1970 to 1994, were obtained from the National Board of Health and Welfare. All patients who underwent amputation after treatment were analyzed separately. Data on deceased patients were extracted from the cause-of-death register. RESULTS: Women had lower survival rates than men at 30 days and 1 and 5 years after operation (89% vs 92%, 75% vs 80%, and 50% vs 55%; P <.001). In the multivariate analysis of risk factors for death during the whole study period, increasing age, calendar year (1980 to 1989), and male sex were significant risk factors (P <.001). The percentage of women who underwent amputation after treatment was comparable with the percentage of men who underwent amputation (11.4% vs 10.2%, P =.075). The multivariate analysis identified increased age and calendar year (1985 to 1994) as important risk factors for amputation (P <.001). Female sex was not found to increase the risk for amputation. Patients who underwent amputation were older than patients with intact limbs (73 vs 68 years, P <.001). The highest percentage of patients who underwent amputation was found among those treated with femorodistal bypass (20%, P =.001). CONCLUSIONS: Female sex is not an important risk factor for poor outcome after treatment for lower limb ischemia, when their increased age has been accounted for. These results imply that we should continue to consider age and type of procedure when estimating outcome, and women should not be refused an intervention on a sex related basis. We should possibly be more selective toward men considering their higher mortality rates. PMID- 11877701 TI - Duplex scanning-guided thrombin injection for the treatment of iatrogenic pseudoaneurysms. AB - PURPOSE: This study presents our current results with duplex scanning-guided thrombin injection (DGTI) for the treatment of lower-extremity iatrogenic pseudoaneurysms (PAs). These results were compared with the results from our patient population that was treated with duplex scanning-guided compression (DGC). METHODS: This was a prospective evaluation of an institutional review board-approved protocol for ultrasound scanning-guided thrombin injection for the treatment of iatrogenic lower-extremity PAs. The maneuver was performed with continuous real-time color ultrasound scanning imaging to guide a needle into a PA sac. Then 0.5 to 1.0 mL of a thrombin solution (1000 U/mL) was injected, and thrombosis of the sac was monitored. All patients underwent an arterial evaluation of the involved extremity before and after thrombin injection. In addition, the size of the PA and its parent artery were documented by means of pre-injection imaging. After thrombosis of the PA, the patient was kept on bed rest for 4 hours, and activity was limited that day (bathroom privileges for inpatients). Follow-up imaging was performed after 24 hours, and attempts were made to obtain imaging 1 week and 1 month after injection. RESULTS: In the 31 months of the study, 131 iatrogenic PAs of the lower extremity were initially treated with DGTI, and thrombosis was achieved in 126 of these cases (96%). Thrombosis of the PA sac was accomplished within seconds of thrombin injection. Five cases failed, three of which resulted from complications of the procedures, with two intra-arterial thrombin injections and one PA rupture after thrombosis. CONCLUSION: Our experience indicates that DGTI is more effective than DGC (96% vs 75%) in the treatment of iatrogenic lower-extremity PAs. The DGTI procedure is completed in minutes, compared with a mean compression time of 44 minutes with DGC, which leads to increased patient and operator acceptance. Intra-arterial thrombin injection was seen in 4% of PAs that were 2.6 cm or smaller and resulted in limb-threatening ischemia requiring surgical intervention. Finally, the use of a biopsy guide attached to the ultrasound scanning transducer head simplifies the visualization of the needle, reducing the number of needle punctures and needle manipulation. PMID- 11877702 TI - Changes in venous lumen size and shape do not affect the accuracy of volume flow measurements in healthy volunteers and patients with primary chronic venous insufficiency. AB - PURPOSE: The purpose of this study was the analysis of the rapid changes in the size and shape of the peripheral vein and the associated changes in blood flow velocities and the estimation of their effect on the reliability of the ultrasound scan volume flow (VF) measurements. METHODS: Ten patients with primary chronic venous insufficiency and 10 healthy volunteers were studied. Two duplex scanners were used simultaneously: one for the velocity measurements in longitudinal plane and another for the cross-sectional area (CSA) measurements in transverse plane during quiet respiration, Valsalva's maneuver (VM), pneumatic cuff compression-decompression, and active dorsiflexion. The patients underwent examination in standing and 15-degrees reverse Trendelenburg's (RT) positions. VF was calculated on the basis of real-time CSA and velocity values. RESULTS: Rapid changes in the CSA as much as 130% for 0.2 seconds were observed. In most cases, the changes in CSA and the flow velocity were inversely related, which resulted in near constant VF. With the exception of VM in the RT position, the difference between real-time VF and mean VF was not significant (P >.05). In the RT position, significant changes in CSA were observed during and immediately after VM. These changes resulted in 23% +/- 15% changes in outflow (both groups) and in 24% +/- 13% changes in reflux (chronic venous insufficiency group). CONCLUSION: The CSA of the peripheral vein and the flow velocities undergo rapid changes during time intervals of a fraction of a second. The vein can have a noncircular cross-section. To minimize the potential error, VF measurements should be performed during quiet respiration or with cuff compression-decompression. With these conditions, the rapid changes in velocities and CSA do not significantly affect the accuracy of VF measurements because of their inverse relation. CSA should be measured planimetrically, or the site of the measurements should be where the vein is close to a circular shape. PMID- 11877703 TI - Reproducibility of ultrasound scan in the assessment of volume flow in the veins of the lower extremities. AB - PURPOSE: The purpose of this study was the identification of the optimal settings of ultrasound scan flow measurement in the veins and the determination of whether the standardization of these settings can provide acceptable reproducibility of the venous flow measurements in individual segments of the lower extremity veins. METHODS: The venous cross-sectional area, the time average mean velocity, and the venous volume flow of 25 healthy volunteers were examined with duplex ultrasound scanning. Reproducibility was examined with different measurement settings. Doppler scan sample volume size, ultrasound scan beam incident angle, and time interval of measurement were varied across a spectrum for arrival at the setting for highest reproducibility of the flow volume measurements. Test-retest reproducibility of venous flow volume measurements then was investigated with optimized settings. RESULTS: The highest repeatability of volume flow measurements was achieved when the full lumen of the vein was insonated (coefficient of repeatability [CR] = 1.88 cm/s), the ultrasound scan beam incident angle was equal to 60 degrees (CR = 1.56 cm/s), and the measurement time was more than 40 seconds (CR = 1.64 cm/s). The mean values of volume flow were 360 mL/min in the common femoral vein, 147 mL/min in the superficial femoral vein, 86 mL/min in the profunda femoral vein, and 38 mL/min in the greater saphenous vein. Test-retest repeatability coefficients were 96.9 mL/min for the common femoral vein, 70.2 mL/min for the superficial femoral vein, 40.8 mL/min for the profunda femoral vein, and 16.8 mL/min for the greater saphenous vein. CONCLUSION: The reproducibility of ultrasound scan measurements of volume flow in veins is optimized with the use of sampling volumes that cover the entire venous lumen, with an incident angle of 60 degrees and measuring for 40-second intervals or longer. With these defined variables, volumetric measurements are sufficiently repeatable. the values of flow volume measured with duplex ultrasound scanning were comparable to those with thermodilution techniques that were reported previously. PMID- 11877704 TI - Differential effects of vascular growth factors on arterial and venous angiogenesis. AB - OBJECTIVE: Angiogenesis, the development of new blood vessels, has become an area of increased interest for both scientific and clinical application purposes. Proangiogenic agents, such as vascular endothelial growth factor (VEGF) and naltrexone, have been shown to effectively induce new blood vessel growth. Other growth factors, such as the endogenous opioid growth factor (OGF; [Met(5)] enkephalin) and retinoic acid, are inhibitors of angiogenesis. The differential effects on veins and arteries, however, by any vascular growth factor, have not previously been investigated. METHODS: The chick chorioallantoic membrane (CAM) assay was used for the in vivo quantitation of angiogenesis. After 3 days of incubation, fertilized chick embryos were explanted, and a 3.2-mm methylcellulose disk containing either the known angiogenic stimulators VEGF (0.2 microg, 1.0 microg) or naltrexone (0.1 microg, 5.0 microg), or the angiogenic inhibitors OGF (1.0 microg, 5.0 microg) or retinoic acid (1.0 microg) was placed onto the CAM surface. An equal volume of distilled water served as a control. After 2 days of growth, the CAM arteries and veins were identified, and images were obtained with a digital camera. Quantitative analysis of angiogenesis was performed on a 100 mm(2) area surrounding the applied disk, and the number and length of the veins and arteries were measured. RESULTS: The angiogenic stimulators VEGF and naltrexone markedly increased both the total number and length of all blood vessels as compared with control values. The mean length of blood vessels decreased, suggesting the induction of new vessel growth. VEGF and naltrexone proportionately increased vein and arterial angiogenesis, maintaining artery/vein ratios for vessel number and length that were unchanged compared with controls. The angiogenic inhibitors, OGF and retinoic acid, notably decreased the total number and length of blood vessels in the CAM preparations. However, these compounds had a disproportionately greater inhibitory effect on arterial angiogenesis as reflected in decreased artery/vein ratios for vessel number and length. CONCLUSIONS: The angiogenic stimulators VEGF and naltrexone induce development of veins and arteries in a proportional manner. In contrast, the angiogenic inhibitors OGF and retinoic acid demonstrated a greater inhibitory effect on arterial as compared with venous angiogenesis. Such differential effects on angiogenesis may be important in both defining mechanisms of action and designing therapeutic interventions. PMID- 11877705 TI - Differential regulation of matrix metalloproteinase activities in abdominal aortic aneurysms. AB - BACKGROUND: The increased synthesis of matrix metalloproteinases (MMPs) by aortic smooth muscle cells (SMCs) is thought to be involved in the etiopathogenesis of abdominal aortic aneurysms (AAAs), but the functional regulation and the activation states of these MMPs remain unclear. In this study, we assessed the expression levels and the functional regulation of several MMPs in the pathogenesis of AAAs. METHODS: Human healthy aorta and AAA specimens were homogenized, and the proteolytic activities of MMP-2 and MMP-9 and of the macrophage metalloelastase (MMP-12) were assessed with zymography. Protein expression of MMP-1, MMP-12, membrane-type 1 MMP (MT1-MMP), tissue inhibitor of MMP 1 (TIMP-1), TIMP-2, TIMP-3, alpha-actin, and beta-actin was analyzed with electrophoresis on sodium dodecyl sulfate gels and immunoblotting. RESULTS: MMP 1, MMP-9, and MMP-12 zymogen levels and proteolytic activities were increased in AAAs when compared with healthy aorta. A severe reduction in alpha-actin- positive vascular SMCs was observed in all the AAA specimens and was correlated with an increase in TIMP-3 but not TIMP-1 or TIMP-2 potential activities. Although pro--MMP-2 activity was decreased, the extent of activated MMP-2 remained unaffected in the AAAs. In accordance with this result, a highly activated MT1-MMP form was also observed in AAAs. CONCLUSION: These data suggest that chronic aortic wall inflammation is mediated by macrophage infiltration, which may account for the destruction of medial elastin, as reflected by SMC down regulation, through increased levels of active MMP-1 and MMP-12. Moreover, altered MT1-MMP proteolytic turnover and differential regulation of TIMP expression in AAAs suggest that tight regulatory mechanisms are involved in the molecular regulation of MMP activation processes in the pathogenesis of AAAs. PMID- 11877706 TI - Pathologic and histologic results of electrical impulses in a rabbit model of atherosclerosis: 24-hour versus 8-hour regimen. AB - OBJECTIVE: Low frequency electrical impulses (EIs) reduce new atherosclerotic plaque formation in previously diseased arteries and may reverse the extent of previous pathologic damage in these structures. METHODS: A pacemaker was implanted on the left side of rabbit abdominal aortas, and an electrode was placed close to the other side of the aorta in the psoas major muscle. For the induction of atherosclerosis, the rabbits were placed on a high cholesterol diet (HCD) for 11 weeks. No EIs were applied to the control series I. In the experimental series, the rabbits were fed an HCD for 3 weeks, after which EIs were applied simultaneously with an HCD for 8 additional weeks (3V, 30 contractions per minute). Experimental series II had 24-hour/day EIs, and series III had 8-hour/day EIs. RESULTS: The closer to the area where the EIs were applied, the more local severity increased (atherosclerosis level and surface area). In the control series, the severity of atherosclerosis in the lower aorta assessed with an arbitrary grading system was 1.75 +/- 0.5 (versus 1.5 +/- 0.57 with 8-hour/day EIs and 0.5 +/- 0.3 with 24-hour/day EIs). The involved surface area was 32.5% +/- 9.5% (versus 1.0% +/- 0.8% with 8-hour/day EIs and 0.75% +/- 0.95% with 24-hour/day EIs). CONCLUSION: Both 24-hour/day and 8-hour/day EIs applied close to the abdominal aorta decreased the severity of atherosclerosis in rabbits placed on a HCD, but 24-hour/day EIs decreased the severity more extensively. PMID- 11877707 TI - Molecular biology: a brief overview. PMID- 11877708 TI - Melioidosis presenting as an infected intrathoracic subclavian artery pseudoaneurysm treated with femoral vein interposition graft. AB - We present the first case of in situ replacement of an infected subclavian artery using superficial femoral vein and the fourth reported case of an infected arterial pseudoaneurysm caused by pseudomonas pseudomallei. Sepsis and hoarseness developed in a 58-year-old man after recent travel to Borneo, Indonesia. Indirect laryngoscopy revealed a paralyzed right vocal cord. Computed tomography and arteriography revealed a 6.5-cm pseudoaneurysm of the proximal right subclavian artery. Blood cultures grew pseudomonas pseudomallei. An abnormal cardiac stress test prompted a coronary angiography, which revealed severe coronary artery disease.The patient underwent coronary artery bypass and in situ replacement of the infected subclavian artery pseudoaneurysm with a superficial femoral vein, along with placement of a pectoralis major muscle flap to cover the vein graft. Operative cultures of the pseudoaneurysm grew pseudomonas pseudomallei. The patient was treated with a 6-week course of intravenous ceftazidime and oral doxycycline and then continued on oral amoxicillin-clavulanate. One week after discontinuing intravenous antibiotics, the patient presented to the emergency department with a rapidly expanding, pulsatile mass in the right supraclavicular space. He was taken emergently to the operating room. After hypothermic circulatory arrest was accomplished, the disrupted vein graft and aneurysm cavity were resected and the subclavian artery was oversewn proximally and distally. Parenteral ceftazidime was continued for 3 months and oral amoxicillin clavulanate (augmentin) was continued indefinitely. There was no evidence of infection clinically or by computed tomographic scan 2 years later. Although autogenous vein replacement of infected arteries and grafts may be successful in the majority of cases, this strategy should probably be avoided when particularly virulent bacteria such as the organism in this case are present. PMID- 11877709 TI - Congenital absence of the external carotid artery: atherosclerosis without a bifurcation. AB - We report the case of a patient with congenital absence of the external carotid artery in whom we performed a carotid endarterectomy. The radiographic features and operative findings are presented. Four similar cases previously reported in the literature are reviewed. A comment on the pathophysiology of atherosclerosis at the carotid bulb in the absence of a bifurcation and a brief discussion on the possible embryologic explanation of this anomaly are discussed. PMID- 11877711 TI - Late endoleak after endovascular repair of an abdominal aortic aneurysm with multiple proximal extender cuffs. AB - Late onset graft or attachment site-related endoleaks may be hazardous, and early identification of patients at risk is important. We describe a patient who underwent implantation of a bifurcated stent graft 5.5 cm below the renal arteries because of a technical error with three extender cuffs implanted proximally to bridge the gap. During the 1st year, aneurysm diameter decreased from 68 to 52 mm. After 1 year, the patient had an acute endoleak develop, which originated between two of the extender cuffs and which was accompanied by severe abdominal pain and reexpansion of the aneurysm. This endoleak was treated with insertion of an additional bifurcated stent graft within the extender cuff segment. The patient has been subsequently followed for 6 months and has had no endoleak or symptoms, and aortic diameter has decreased once again to 55 mm. PMID- 11877710 TI - Arterial infection and staphylococcus aureus bacteremia after transfemoral cannulation for percutaneous carotid angioplasty and stenting. AB - In this report, we present a patient who developed an infected femoral artery after repuncture cannulation for carotid angioplasty and intraluminal stenting. The case was complicated by persistent bacteremia and a delay in diagnosis before it was managed successfully with an autogenous replacement graft and appropriate antibiotics. Overt stent infection is exceedingly rare, but according to the literature describing transfemoral coronary artery intervention, the spectrum of clinical syndromes related to infection of the arterial puncture site includes local invasion, pseudoaneurysm formation, septic embolization to the distal limb, and bacteremia. The diagnosis requires a high degree of clinical suspicion and is often delayed. Although the incidence of infectious complications reported for percutaneous intra-arterial interventions historically has been low, the absolute number of these complications almost certainly will increase in the future because of the expanding array of interventional procedures that is becoming available. PMID- 11877712 TI - Endovascular management of iliac limb occlusion of bifurcated aortic endografts. AB - Although aortic endograft iliac limb occlusion is an uncommon event, its treatment is problematic because standard surgical thrombectomy risks graft dislodgment or component separation. Although femorofemoral bypass grafting can restore perfusion to the affected limb, its longevity may be inferior to reestablishing patency of the endograft itself and represents a failure of the endograft procedure. With aortic endografts now commercially available, implanting surgeons must be aware of this important complication and well versed in all of the endovascular treatment options. We report three cases of endoluminal management of unilateral iliac limb occlusion of bifurcated aortic endografts. PMID- 11877713 TI - Common femoral artery injury secondary to bicycle handlebar trauma. AB - Blunt trauma from bicycle handlebars is associated with well-described injuries of the abdominal viscera. These injuries result from the forceful compression of the relatively immobile abdominal organs between the handlebar end and the vertebral bodies. The common femoral artery is also immobile as it passes anterior to the superior pubic ramus, rendering this vessel susceptible to a similar mechanism of injury. We have treated two children who sustained thrombosis of the common femoral artery caused by bicycle handlebar trauma. The lack of familiarity with this uncommon mode of injury may contribute to delayed diagnosis and increased morbidity. We therefore wish to draw attention to this mechanism of injury. PMID- 11877714 TI - Percutaneous bedside femorofemoral bypass grafting for acute limb ischemia caused by intra-aortic balloon pump. AB - Acute limb ischemia is a common, recognized complication of intra-aortic balloon pump (IABP) placement in patients with failing myocardium, and an operative femorofemoral bypass graft is often necessary in IABP-dependent patients as a means of maintaining the lower-limb perfusion. In this report, we present a minimally invasive endovascular technique for creating a percutaneous temporary femorofemoral bypass graft at bedside in patients with IABP-induced limb ischemia. This temporary bypass grafting technique may obviate a potential graft infection or wound complications associated with a formal femorofemoral bypass graft. PMID- 11877715 TI - Regarding: "The incidence and natural history of type I and II endoleak: a 5-year follow-up assessment with color duplex ultrasound scan". PMID- 11877716 TI - Percutaneous bioprosthetic venous valve: a long-term study in sheep. AB - A long-term evaluation of a new percutaneously placed bioprosthetic, bicuspid venous valve (BVV) consisting of a square stent and small intestinal submucosa (SIS) covering was performed in 12 sheep. Of 26 BVVs placed into the jugular veins, 25 exhibited good valve function on immediate venography and 22 on venograms obtained before the sheep were killed. Gross and histologic examination results demonstrated incorporation of remodeled and endothelialized SIS BVVs into the vein wall. Slight to moderate leaflet thickening was found mostly at their bases. Percutaneously placed SIS BVV is a promising one-way, competent valve that resists venous back-pressure while allowing forward flow. PMID- 11877717 TI - Recommended standards for reports dealing with arteriovenous hemodialysis accesses. AB - The incidence rate of treated end-stage renal disease in the united states is 180 per million and continues to rise at a rate of 7.8% per year. Arteriovenous hemodialysis access (AV access) creation and maintenance are two of the most difficult issues associated with the management of patients on hemodialysis. The 1-year complication rate of a primary prosthetic AV access for hemodialysis ranges from 33% to 99%. Various investigators report on patency and complications of AV access. However, it is rather difficult to compare outcomes because of the wide variety of access materials, configurations, locations, risk factors, and quality of inflow and outflow vessels. Although there have been reporting standards for dialysis access endovascular interventions and for central venous access placement, standards regarding surgical access placement and its revision are lacking. The "Dialysis Outcome Quality Initiative," published by the National Kidney Foundation, provides recommendations for optimal clinical practices aimed at improving dialysis outcome and patient survival. This reporting standards document is not meant to be a "practice guidelines" or "best practices" document. Rather, the purpose of this document is to provide standardized definitions related to AV access procedures and to recommend reporting standards for patency and complications, to be used by surgeons, nephrologists, and interventional radiologists, that will permit meaningful comparisons among AV access procedures. The terms, definitions, and categories featured in this article have been approved by the Committee on Reporting Standards of the Society for Vascular Surgery and the American Association for Vascular Surgery and should be observed in preparing manuscripts on AV accesses for submission to the Journal Of Vascular Surgery. PMID- 11877718 TI - Transluminal repair of abdominal aortic aneurysm: a call for selective use, careful surveillance, new device design, and systematic study of transrenal fixation. PMID- 11877719 TI - Lifeline Registry of Endovascular Aneurysm Repair: Registry data report. AB - PURPOSE: The goal of the Lifeline Endovascular Registry is to provide a minimal, yet comprehensive, data set of patient follow-up that can evaluate the long-term safety of endovascular grafts used in abdominal aortic aneurysm repair. METHODS: Follow-up data have been collected on 1757 patients for this first report, including 1646 endovascular graft recipients and 111 surgical patients receiving treatment for abdominal aortic aneurysm. RESULTS: Logistic regression of 1-year survival indicates that the factors most likely to decrease 1-year survival for the surgical group are renal failure and larger aneurysm size. For the endovascular graft recipients, the presence of renal failure, chronic obstructive pulmonary disease, congestive heart failure, larger aneurysm size, and increased age demonstrates a decrease in 1-year survival. Approximately 80% (1309) of the endovascular recipients have been followed for 1 year. Of these 1309 patients, 17% (222) have incurred an endoleak, and enlargement of the aneurysm has occurred in 4.6% (60). There was a total of 80 (4.9%) endovascular graft recipients converted to surgery. The most prevalent factors causing conversion to surgery were an increase in aneurysm diameter, aneurysm rupture, and the presence of a proximal endoleak. CONCLUSIONS: Although the data collected by the Registry thus far are limited, we anticipate rapid expansion of the Registry to include data from other manufacturers and investigators. Nevertheless, early data analysis does demonstrate the importance of surveillance of endovascular graft recipients. Through the collection and analysis of Registry data, adverse events continue to be captured, enabling the monitoring of long-term safety of endovascular grafts and the evaluation of graft performance. Data on comorbidities and postoperative factors, collected and evaluated for their effect on survival of the graft recipients, also provide information on optimal patient selection and management. We expect that the next report will support and expand on these findings, providing continuing evidence of the value of a national endovascular registry. PMID- 11877720 TI - Abdominal aortic aneurysm in death row inmate. PMID- 11877721 TI - Regarding "Heparin-bonded Dacron or polytetrafluoroethylene for femoro-popliteal bypass grafting: a multicenter trial". PMID- 11877723 TI - Regarding "The 50th anniversary of abdominal aortic reconstruction". PMID- 11877724 TI - Regarding "The influence of surgical specialty training on the outcomes of elective abdominal aortic aneurysm surgery". PMID- 11877727 TI - Regarding "Aneurysm sac pressure measurements after endovascular repair of abdominal aortic aneurysms". PMID- 11877728 TI - Regarding "A comparison of para-anastomotic compliance profiles after vascular anastomosis: nonpenetrating clips versus standard sutures". PMID- 11877730 TI - The use of a nerve stimulator in difficult carotid surgery. PMID- 11877731 TI - [Idiopathic pulmonary fibrosis: adverse prognosis in artificial respiration patients]. PMID- 11877732 TI - [A novelty: Joint Congress of DGP and DGAI]. PMID- 11877733 TI - [On the practice of patient training in obstructive diseases of the respiratory tract (asthma and COPD) in Germany]. PMID- 11877734 TI - [In-patient asthma education and self-management training in clinical practice--a national survey in Germany]. AB - BACKGROUND: According to international guidelines patient education is a key component of modern asthma-management. Especially in rehabilitation, patient training is considered essential for long-term treatment success. Based on growing empirical evidence for clinical efficacy of asthma self-management training, standardized education programs as well as guidelines and recommendations for quality management in health care practice have been developed. AIM: Due to the lack of evidence [empirical data] on compliance to these recommendations we conducted a nation-wide survey including in-patient pneumological clinics in Germany. METHODS: Exploiting address registers of national asthma organizations (i. e. Deutsche Atemwegsliga) we identified 75 clinics that offer education programs on a regular basis, 65 of which participated in this study (drop-out-rate: 12 %). These institutions were asked to complete a questionnaire in order to assess various aspects of their education practice. RESULTS: Although 91 % of clinics reported to adhere to guidelines and recommendations for patient education, merely 50 % supplied standardized and evaluated education and self-management training programs. A rather small proportion (19 %) was found to fully adhere to protocol (maximum treatment integrity, no mixing of different education schemes). Furthermore, substantial variance between clinics was observed in actual performance of asthma education, the major indicators being: indication criteria and eligibility of patients; education contents; group size, duration and frequency of sessions; extent and intensity; didactical methods; number and qualification of trainers; measures of quality management, and settings. Accordingly, a consistent standard of patient education in rehabilitation practice has not yet been achieved. CONCLUSION: While substantial advances in the proposition of patient education and self-management trainings in in-patient health care remain unquestioned, there are some deficits in actually performing such programs in clinical practice. Results also show that another question needs to be addressed: qualifying trainers. Less than half of trainers participating in this study reported to have absolved a train-the trainer-course. PMID- 11877735 TI - [Investigation of growth prognosis in asthmatic children]. AB - The objective of the present study was the ascertainment of correct data related to growth prognosis in asthmatic children. For this purpose, 210 young asthmatics including 153 males and 57 females ranging in age from 1.7 to 18.9 years were evaluated. Data were specified for the parameters: chronological age (CA); bone age (KA); height age (LA); height (KH); severity of illness; target adult height (GZG) and predicted adult height (PEG). Among this group, 5.9 % of the boys and 3.5 % of the girls, were found to be of small stature (KH < - 2 s). This corresponds to about 2.5 and 1.5 times the average in a normal distribution. Children with the highest severity of illness showed the strongest negative deviation for the KH-SDS values (boys - 0.59 plus minus 1.1, girls - 0.97 plus minus 0.8). This proved to be statistically significant for females. Development of CA, KA and LA continued unremarkably until the age of 4 years. From age 5, partially a significant growth retardation of KA and LA could be observed in both sexes. To age 16, KA values and, after that LA values, were more strongly affected. Comparison of the GZG and PEG average values within the degree of severity groups made it clear that the PEG values for both sexes, independent of the severity of illness, were without exception, smaller than the GZG. The differences of both parameters proved to be statistically significant for the group with the most severe symptoms of the boys (180.8 plus minus 6.5 cm to 175.8 plus minus 6.2 cm, p < 0.01) and for the entire groups of both sexes (boys 180.2 plus minus 5.3 cm to 178.6 plus minus 7.3 cm, p < 0.05, girls 167.7 plus minus 4.4 cm to 165.8 plus minus 6.8 cm, p < 0.05). However, a separate comparison of GZG and PEG average values, respectively, among the severity groups showed no significant differences. The results argue for a growth-inhibiting influence from bronchial asthma. The cause has to be severity of illness but direct effects of the atopy on skeletal development is also taken in consideration. Confirmation of these findings requires investigation of a larger group of asthmatics and clarification of the pathophysiological processes in the growing skeletons of atopic children. PMID- 11877736 TI - [Occupational airway sensitization due to subtilisin]. AB - Since the 1960s an increasing number of occupational allergies against natural, and against modified recombinant enzymes were observed. Subtilisin, a frequently used enzyme often has been shown to cause airway sensitization. The present evaluation of clinical literature data confirms the sensitization potential of subtilisin. Several cases of specific airway hyperresponsiveness caused by subtilisin are verified by a number of studies. As symptoms, results of skin prick tests, detection of specific IgE-antibodies and results of specific bronchoprovocation tests are consistent, an immunologic mechanism is confirmed. PMID- 11877738 TI - [Evolutionary quality control of diagnostics and therapy in hospitals]. PMID- 11877737 TI - [Guidelines for conducting provocation tests with allergens. German Society of Allergology and Clinical Immunology and German Society for Immunology]. AB - Guidelines for bronchial allergen provocation tests, first published in 1984, are updated by this paper. This version has been compiled in repeated meetings of a panel of experts after evaluation of the publications available. Precise statements regarding indications, contraindications, after safety measures are given. Three evidence-based protocols are submitted in detail. PMID- 11877739 TI - Influence of compound structure on affinity, sequence selectivity, and mode of binding to DNA for unfused aromatic dications related to furamidine. AB - In the course of a program aimed at developing sequence-specific gene-regulatory small organic molecules, we have investigated the DNA interactions of a new series of nine diphenylfuran dications related to the antiparasitic drug furamidine (DB75). Two types of structural modifications were tested: the terminal amidine groups of DB75 were shifted from the para to the meta position, and the amidines were replaced with imidazoline or dimethyl-imidazoline groups, to test the importance of both the position and nature of positively charged groups on DNA interactions. The interactions of these compounds with DNA and oligonucleotides were studied by a combination of biochemical and biophysical techniques. Absorption and CD measurements suggested that the drugs bind differently to AT and GC sequences in DNA. The para-para dications, like DB75, bind into the minor groove of poly(dAT)(2) and intercalate between the base pairs of poly(dGC)(2), as revealed by electric linear dichroism experiments. In contrast, the meta-meta compounds exhibit a high tendency to intercalate into DNA whatever the target sequence. The lack of sequence selectivity of the meta-meta compounds containing amidines or dimethyl-imidazoline groups was also evident from DNase I footprinting and surface plasmon resonance (SPR) experiments. Accurate binding measurements using the BIAcore SPR method revealed that all nine compounds bind with similar affinity to an immobilized GC sequence DNA hairpin but exhibit very distinct affinities for the corresponding AT hairpin oligonucleotide. The minor groove-binding para-para compounds have a high specificity for AT sequences. The biophysical data clearly indicate that shifting the cationic substituents from the para to the meta position results in a loss of specificity and change in binding mode. The strong AT selectivity of the para para compounds was independently confirmed by DNase I footprinting experiments performed with a range of DNA restrictions fragments. In terms of AT selectivity, the compounds rank in the order para-para > para-meta > meta-meta. The para dications bind preferentially to sequences containing four contiguous AT base pairs. Additional footprinting experiments with substrates containing the 16 possible [A.T](4) blocks indicated that the presence of a TpA step within an [A.T] (4) block generally reduces the extent of binding. The diverse methods, from footprinting to SPR to dichroism, provide a consistent model for the interactions of the diphenylfuran dications with DNA of different sequences. Altogether, the results attest unequivocally that the binding mode for unfused aromatic cations can change completely depending on substituent position and DNA sequence. These data provide a rationale to explain the relationships between sequence selectivity and mode of binding to DNA for unfused aromatic dications related to furamidine. PMID- 11877740 TI - Preferred conformations of RGDX tetrapeptides to inhibit the binding of fibrinogen to platelets. AB - The conformational study on Arg-Gly-Asp (RGD)-containing tetrapeptides in the unhydrated and hydrated states has been carried out using the force field ECEPP/3 and the hydration shell model. The tetrapeptides studied here are H-RGDX-OH (X = Trp, Tyr, Phe, Leu, Val, Cys, Gln, and Ser), which show the inhibitory activity for binding of fibrinogen to platelets in the order of RGDW approximately equal to RGDY approximately equal to RGDF approximately equal to RGDL > RGDV > or = RGDC > or = RGDQ > or = RGDS. The backbone conformations with two C(7) backbone to-backbone hydrogen bonds between Asp and Arg residues and between Xaa and Gly residues are in common most probable for the RGD sequence of RGDX tetrapeptides in the hydrated state. The dominant beta-turns for RGDX are found to be the types V' and IV at Gly-Asp and Asp-Xaa sequences, respectively, which are quite similar to the types II' and I (or II), respectively. However, it cannot be ruled out that the extended conformations are also remarkably feasible for RGDX tetrapeptides in water by peering the distributions of backbone conformations. These calculated results are consistent with the experimental results on RGD containing proteins and conformationally constrained RGD-containing peptides. The reason why the RGDX becomes more potent as the side chain of the X residue is more hydrophobic may be ascribed to that the more hydrophobic is the residue X, the more populated are beta-turn structures for the Gly-Asp sequence. The hydrophobic side chain of X residue exposed to water is likely to interact with the hydrophobic region of receptor easily. PMID- 11877741 TI - Synthesis, conformational analysis, and spectroscopic characterization of peptides based on Daf, the first rigid transition-metal receptor, cyclic C(alpha,alpha)-disubstituted glycine. AB - Three series of terminally protected model oligopeptides to the nonamer level, based on 9-amino-4,5-diazafluorene-9-carboxylic acid, the first rigid bipyridine type C(alpha,alpha)-disubstituted glycine, and either Gly, L-Ala, or Aib residues were synthesized by solution methods and fully characterized. The molecular structures of two derivatives and one tripeptide were determined in the crystal state by x-ray diffraction. Moreover, the solution preferred conformations of these peptides were assessed by Fourier transform infrared absorption and (1)H NMR techniques. A comparison with the known structural tendencies of the strictly related C(alpha,alpha)-disubstituted glycyl residues 1-aminocyclopentane-1 carboxylic acid and 9-aminofluorene-9-carboxylic acid is made, and the implications for the use of the 9-amino-4,5-diazafluorene-9-carboxylic acid residue in conformationally constrained analogs of bioactive peptides are briefly examined. A spectroscopic (uv absorption, fluorescence, CD) characterization of this novel heteroaromatic C(alpha,alpha)-disubstituted glycine is also reported. Finally, preliminary conformational data and membrane activity measurements are discussed for an analog of the lipopeptaibol antibiotic [L-Leu(11)-OMe] trichogin GA IV in which a 9-amino-4,5-diazafluorene-9-carboxylic acid residue was synthetically incorporated in position 1 (replacing the original Aib residue). PMID- 11877742 TI - Interaction between Btk TH and SH3 domain. AB - Several mechanisms are involved in the regulation of cellular signaling. Bruton tyrosine kinase (Btk) of the Tec family contains in the Tec homology (TH) domain a proline-rich region (PRR) capable of interacting with several SH3 domains. The Btk has the SH3 domain adjacent to the TH domain. CD and fluorescence spectroscopy were used to study the binding of two peptides corresponding to segments in the PRR to the Btk SH3 domain. The peptide for the N-terminal half of the PRR binds specifically, whereas the other peptide had hardly any affinity. The TH domain has about four times lower affinity to the SH3 domain than the peptide, 17.0 vs 3.9 microM. The interaction was further tested with an SH3 domain construct that contained the PRR. The two peptides cannot compete for the binding to the extended protein and the TH domain has two times lower affinity to the extended SH3 domain. The intra- or intermolecular interaction between the TH and SH3 domain might have regulatory function also in the other Tec family members. PMID- 11877743 TI - Analysis of peptide rotational diffusion by homonuclear NMR. AB - The analysis of the rotational diffusion of a molecule using homonuclear NMR is investigated. The homonuclear longitudinal and transverse cross-relaxation rates, which can be quantitatively measured using off-Resonance Rotating frame nuclear Overhauser Effect Spectroscopy (ROESY), are used to build a distribution, which exhibits a solid-state-like pattern characteristic of the diffusion tensor. The distributions of the antimicrobial peptide ranalexin in water and in 30% of trifluoracetic acid (TFE) are compared, and the peptide rotational diffusion is shown to be more isotropic in water than in 30% TFE. This difference is further supported by the analysis of NMR ranalexin conformers in 30% TFE, and by the analysis of a molecular dynamics simulation of peptide in water. PMID- 11877744 TI - Eighth Conference on Radioimmunodetection and Radioimmunotherapy of Cancer. Introduction. PMID- 11877745 TI - Alpha-melanocyte-stimulating hormone peptide analogs labeled with technetium-99m and indium-111 for malignant melanoma targeting. AB - BACKGROUND: Previous studies have shown that the compact structure of a rhenium cyclized alpha--melanocyte-stimulating hormone peptide analog, [Cys3410,D Phe7]alpha-MSH(3--13), or Re-CCMSH, significantly enhanced its in vivo tumor uptake and retention. In this study, the metal chelate 1,4,7,10 tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA) was coupled to the N terminus of Re-CCMSH in order to develop a melanoma-targeting peptide that could be labeled with a wider variety of imaging and therapeutic radionuclides. METHODS: Biodistribution properties of indium-111 ((111)In)--labeled DOTA-Re CCMSH were compared with the non-DOTA-containing technetium-99m ((99m)Tc)--CCMSH in murine melanoma--bearing C57 mice to determine the effects of DOTA on tumor uptake and whole-body clearance. The tumor targeting capacity and clearance kinetics of (111)In-DOTA-Re-CCMSH were also compared with other related cyclic and linear (111)In-labeled DOTA-alpha-MSH complexes. RESULTS: The in vivo distribution data showed that the conjugation of DOTA to Re-CCMSH did not reduce its initial tumor uptake kinetics but did enhance its tumor retention and renal clearance properties. The tumor uptake of (111)In-DOTA-Re-CCMSH was significantly higher than the other (111)In-DOTA--coupled cyclic or linear alpha-MSH analogs used in this study. Moreover, (111)In-DOTA-Re-CCMSH displayed lower radioactivity accumulation in normal tissues of interest than its non-Re-cyclized counterpart, (111)In-DOTA-CCMSH; the disulfide bond--cyclized (111)In-DOTA-CMSH; or the linear (111)In-DOTA-NDP. CONCLUSIONS: Peptide cyclization via rhenium coordination significantly enhanced the tumor targeting and renal clearance properties of DOTA Re-CCMSH, making it an excellent candidate for melanoma radiodetection and radiotherapy. PMID- 11877747 TI - Apoptosis induced by low-dose and low-dose-rate radiation. AB - BACKGROUND: It has been claimed that external radiation, as a treatment modality for malignant diseases, partly induces apoptosis. It is not known, however, whether therapeutic low-dose and low-dose-rate radiation are able to induce apoptosis. METHODS: The effect of low-dose radiation on apoptosis induction in HeLa Hep2 cells was studied, and quantitation of the apoptotic cells was performed by immunocytochemistry using TdT-mediated dUtp-x Nick End Labeling (TUNEL) technology and the M30 CytoDEATH antibody method. RESULTS: When TUNEL staining was used to quantify apoptosis in untreated HeLa Hep2 cells kept in culture, approximately 5 plus minus 3% of the cells showed positive staining without any treatment. In the first experiment, the HeLa Hep2 cells were exposed to gamma radiation (i.e., 0.5, 1, 2, 5, 10, and 15 grays [Gy]) from a cobalt-60 radiation source delivering a dose rate of 0.80 Gy/min. The radiated cells were cultivated for 5, 10, 24, 48, 72 and 168 hours after irradiation. Radiation doses below 2 Gy did not cause any significant apoptosis, but between 5 and 15 Gy significant apoptosis was observed, with peak values at 5 Gy (P < 0.001). Up to 60% of the investigated cells were shown to display apoptosis. Time to this peak value was 168 hours after irradiation. The HeLa Hep2 cells were exposed to doses of 2, 5, and 10 Gy at a 10-fold lower dose rate (0.072 Gy/min). The cells that achieved a dose below 2 Gy did not present increased apoptosis. At doses above 2 Gy, however, the cells again demonstrated significant apoptosis. Up to 24 hours following irradiation, no apoptosis could be documented, whereas beyond 24 and up to 168 hours a highly significant apoptosis induction was observed. Significant cytotoxicity was confirmed by chromium-51 release from the cells at 5 Gy. CONCLUSIONS: Low-dose and low-dose-rate radiation are able to induce significant apoptosis, and apoptosis may be one of the mechanisms by which low-dose radiation causes growth inhibition. PMID- 11877746 TI - Comparison of the biologic effects of MA5 and B-B4 monoclonal antibody labeled with iodine-131 and bismuth-213 on multiple myeloma. AB - BACKGROUND: Using a specific monoclonal antibody (MAb), B-B4, coupled to bismuth 213 ((213)Bi) by a chelating agent (CITC-DTPA), the feasibility of alpha radioimmunotherapy (RIT) for multiple myeloma (MM) has been demonstrated previously. METHODS: In this study, the two MAbs tested, MA5 and B-B4, target the epithelial antigens Muc-1 and syndecan-1, respectively, which are both expressed by MM cell lines. Antibody characterization was evaluated by flow cytometric analysis of normal and tumoral hematopoeitic cells of MM patients as well as immunohistochemical tests of normal, nonhematopoetic tissues. Radiobiologic effects were evaluated for (213)Bi- and iodine-131 ((131)I)--labeled antibodies. We assessed in vitro mortality (thymidine incorporation, MTT, and clonogenic assays) and cell cycle modifications with propidium iodide staining. These tests were performed on MM cell lines until 120 hours postirradiation at several time points, using radiolabeled antibody concentrations ranging from 0.5 to 20 nM and specific activities ranging from 240 to 1200 MBq/mg of MAb. RESULTS: MA5 stained all MM cells in only 50% of patients, whereas B-B4 recognized all MM cells in all patients. B-B4 principally showed hepatic, pulmonary, and duodenal staining, whereas MA5 marked renal and pulmonary tissues. RIT with (213)Bi-B-B4 induced specific mortality and G(2)/M phase cell cycle arrest, which depended on the concentrations and specific activity. For (213)Bi-MA5, this arrest appeared at concentrations above 10 nM, an amount fivefold higher than that required with B B4. This difference was also found in thymidine incorporation assays. Furthermore, with (213)Bi-B-B4, the arrest at the G(2)/M phase appeared quickly, within 24 hours after irradiation, and affected up to 60% of the cells (for 20 nM of (213)Bi-B-B4 at 1,200 MBq/mg). Conversly, (131)I-B-B4 had a very limited effect on cell mortality and did not induce any cell cycle arrest. CONCLUSIONS: The results of this study show that B-B4 might be the more effective therapeutic antibody and suggest that alpha-RIT might be more suitable than beta-RIT for treating single-cell tumor models. Thus, these findings set the stage for the beginning of clinical trials using alpha-emitter--radiolabeled B-B4, with special attention paid to hepatic, pulmonary, and intestinal side effects. PMID- 11877748 TI - Radionuclide-antibody conjugates for single-cell cytotoxicity. AB - BACKGROUND: Radioimmunotherapy has primarily utilized high-energy beta-particles, which are intended to kill macroscopic tumor masses. Such conjugates do not kill single cells or micrometastases efficiently. For killing single cells, it may be preferable to use radiation with a much shorter path length, such as alpha particles or Auger or conversion electrons. METHODS: This selective review focuses on the use of radiolabeled antibody (Ab) conjugates to achieve single cell kill. The advantages and disadvantages of particular types of radionuclides, the significance of intracellular localization of the Ab, and the potential clinical application of this approach are discussed. Potentially useful radionuclides are listed. RESULTS: Auger and conversion electrons can kill cells effectively, with at least 6 logs of cell kill. Abs on the cell surface are only slightly less potent than Abs internalized into the cytoplasm, and this is consistent with theoretical considerations. alpha-Particles kill single cells very effectively, but the short half-lives of the available alpha-particle emitters are probably a disadvantage. High-energy beta-particles can also kill single cells if they bind in sufficient amounts, but their disadvantage appears to be greater nonspecific toxicity. CONCLUSIONS: Single-cell kill can be obtained with radionuclide-Ab conjugates. The selection of the optimal radionuclide may depend on the details of the clinical situation, such as the size and accessibility of the tumor burden, and the particular Ab to be used. Direct comparisons of various radionuclides are required in order to identify the optimal approach. However, for single-cell kill, as required for therapy of micrometastases, the use of Auger and conversion electron emitters appears to have substantial advantages. While current methods limit the applicability of this approach to Abs having a high level of binding, it may be applicable to lower-density antigens if higher specific activities or more potent radionuclides can be used. Tumor cure may require a mixture of radionuclides intended to kill both single cells and large tumor masses. PMID- 11877749 TI - Comparison of four scatter correction methods for patient whole-body imaging during therapeutic trials with iodine-131. AB - BACKGROUND: Activity in regions of interest can be measured using serial whole body scintigraphic images to estimate the dose received by a patient after therapeutic injections. As scatter and attenuation introduce biases in quantitative measurements, these phenomena need to be corrected to allow accurate determination of tracer concentration. METHODS: The feasibility of iodine-131 whole-body imaging in list mode was studied over an extended spectrum (0-750 keV) in order to compare four scatter correction methods by the geometric mean approach (20%, Dual Energy Window, Triple Energy Window, and Spectral Factor Analysis methods). All data were corrected for attenuation using a Transmission Attenuation Correction prototype from Sopha Medical Vision international. The half-life of an iodine-131 standard source was calculated from scatter-corrected anterior views. Whole-body activities, using the Day 0, Hour 1, image as a reference (calibration from an administered dose) and an external calibration source (calibration from an imaged known-activity source), were calculated for three patients undergoing a radioimmunotherapy trial in order to assess the reliability of quantification by the geometric mean approach. RESULTS: Patient studies confirmed the clinical feasibility of this type of acquisition. As expected, all methods allowed determination of an accurate half-life for the calibration source. A slight impact of scatter correction was observed in quantification with calibration from an administered dose. For quantification with calibration from an imaged known-activity source, whole-body activity was overestimated by +100% to +200% with the 20% window, depending on the size of the patient, whereas errors were about +50% with scatter correction. However, the influence of patient morphology was less marked when a scatter correction method was used. CONCLUSIONS: When the geometric mean approach is used together with a sophisticated transmission acquisition device for quantification with calibration from an administered dose, the 20% energy window appears to be adequate. However, for quantification with calibration from an imaged known-activity source, accurate activity estimates cannot be obtained even when scatter correction is used to compensate for the influence of patient morphology. PMID- 11877750 TI - The two types of correction of absorbed dose estimates for internal emitters. AB - BACKGROUND: Two types of correction for absorbed dose (D) estimates are described for clinical applications of internal emitters. The first is appropriate for legal and scientific reasons involving phantom-based estimates; the second is patient-specific and primarily intended for radioimmunotherapy (RIT). METHODS: The Medical Internal Radiation Dose (MIRD) relationship (D) = S A is used, where S is a geometric matrix factor and A is the integral of source organ activities. Internal consistency of the data and the size of organ systems in the humanoid phantom must be assured in both types of estimation. RESULTS: The first dose estimate correction (I) is one whereby computations refer to one or another standard (e.g., MIRD-type) phantom. In this case the S value remains as given, but the measured patient A data must be standardized. The correction factor is the phantom's ratio of organ mass to whole-body mass divided by the same ratio for the volunteer or patient. The second dose estimate correction (II) is patient specific. While the A value is unchanged for this application, a correction term is provided for the phantom-derived S matrix. The dominant (nonpenetrating radiation) component of this correction factor can be obtained via the ratio of the patient to phantom organ masses. In both corrections, we recommend that true organ sizes, necessary in each method of estimation, be determined in a separate sequence of anatomic images. CONCLUSIONS: In both dose estimation corrections, true sizes of the patient's or volunteer's internal organs must be obtained. Correction due to organ mass size can be severalfold and is probably the dominant uncertainty in the internal emitter absorbed dose calculation process. PMID- 11877751 TI - Time dependence of the activity concentration ratio of red marrow to blood and implications for red marrow dosimetry. AB - BACKGROUND: The method for red marrow dosimetry in radioimmunotherapy, in the absence of specific activity uptake in red marrow, is based on the activity measured in the blood or plasma. The activity concentration ratio of red marrow to blood is then assumed to be constant. The aim of the current study was to determine whether this ratio varies with time after injection. METHODS: Measurements were carried out with both animals and patients.Tumor-bearing rats were intravenously injected with iodine-131-, iodine-125-, indium-111-, or rhenium-188-labeled BR96, a chimeric immunoglobulin G1 monoclonal antibody. (All were chelate-labeled, except for iodine-131, which was iodogen-labeled.) Measurements were made of the activity concentration in blood and bone marrow at different points in time after injection, and the ratio of activity concentration in red marrow and blood as a function of time postinjection (RMBLR[t)]) was calculated. For patients treated with iodine-131-labeled monoclonal antibody (LL2, Immunomedics Inc., Morris Plains, NJ; anti-CD22; immunoglobulin G2 isotype of mouse origin), blood samples were drawn and scintillation camera images taken at different times after injection. The red marrow activity concentration in the sacrum was determined by activity quantification from regions of interest. The activity concentration in blood was also measured. The RMBLR(t) was calculated based on these data. RESULTS: For both patients and rats, the RMBLR(t) was increased 72 hours after injection. Furthermore, it was found that the use of a constant RMBLR can lead to an over- or underestimation of the absorbed dose in bone marrow. CONCLUSIONS: These data demonstrate the difficulty in using fixed values of the activity concentration ratio of red marrow to blood for dosimetry. PMID- 11877752 TI - Biokinetics of yttrium-90--labeled huBrE-3 monoclonal antibody. AB - BACKGROUND: This study reports summary biokinetics for 17 patients treated with huBrE-3 antibody labeled with indium-111 ((111)In) and yttrium-90 ((90)Y) in a Phase I dose escalation trial. METHODS: Patients were infused with huBrE-3 antibody conjugated to 1-p-isothiocyanatobenzyl 3-methyl- and 1-p isothiocyanatobenzyl 4-methyl-diethylenetriamine pentaacetic acid (MX-DTPA). The huBrE-3 was labeled with increasing amounts of (90)Y radioactivity according to the following activity regimen: 10 mCi/m(2), 20 mCi/m(2), 33 mCi/m(2), 50 mCi/m(2), and 70 mCi/m(2). In addition to the (90)Y activity, 3--5 mCi of (111)In was labeled to huBrE-3 to serve as an imaging agent. In characterizing the biokinetics of huBrE-3, serial urine and blood samples were acquired. Additionally, whole-body imaging using a scintillation camera was performed at four time points postinfusion. RESULTS: Cumulative urine data yielded a plot of total-body biologic excretion that was relatively flat. Total body regions of interest derived from nuclear medicine scintigrams decreased according to a monoexponential function with a slope slightly greater than the rate of physical decay. When physical decay was combined with the urine biologic excretion rate, the calculated rate of activity decrease was indistinguishable from the scintigraphic rate of decrease in total-body activity. CONCLUSIONS: The authors concluded from these observations that the radioactivity remains essentially inside the patient, that biologic excretion of activity from the total body is not appreciable, and that the path for biologic excretion of activity that does occur is via the urine. The half-time associated with the beta (slow) phase for extraction from the blood averages 40.5 hours. Since large amounts of radioactivity do not appear in the urine, and total-body activity is decreased approximately according to physical decay (64.1 hours), activity must pool elsewhere after leaving the blood. The logical place is the skeleton, with possible selective binding to the bone marrow. Bone marrow biopsies from 4 of 7 patients who consented to serial biopsies were supportive of this conclusion. PMID- 11877753 TI - A model-based approach for the optimization of radioimmunotherapy through antibody design and radionuclide selection. AB - BACKGROUND: The effectiveness of radioimmunotherapy (RIT) is known to depend on at least six factors: total absorbed dose and pattern of delivery, radiosensitivity, rate of repair of sublethal damage, ongoing proliferation during treatment, tumor heterogeneity, and tumor size. The purpose of this study was to develop a mathematic model that would relate the absorbed dose and its pattern of delivery to tumor response by incorporating information on each factor. This model was used to optimize therapeutic efficacy in mice by matching the antibody and radionuclide characteristics while ensuring recoverable marrow toxicity. METHODS: Pharmacokinetic data were acquired in mice for a range of antibodies that varied in molecular weight, specificity, affinity, and avidity, and for a range of tumor sizes. This information was combined with the properties of iodine-131, rhenium-86, and yttrium-90 to determine the pattern of dose delivery. Tumor response was characterized in terms of radiosensitivity, rate of repair, and proliferation. Values for these parameters were obtained from in vitro assays and were incorporated into a response model based on the linear quadratic model. Storage phosphor plate technology was used to acquire images of antibody distribution in tumor sections. These were registered with corresponding images showing tumor morphology, which were subsequently used to delineate regions that were distinct in terms of their response to radiation: oxygenated, radiosensitive areas that contained viable cells and hypoxic areas containing resistant viable cells and necrotic cells. Beta point dose kernels were then used to estimate the absorbed dose distribution in these regions. RESULTS: Therapy in normoxic areas was more effective than in hypoxic areas. The multivalent, tumor specific antibodies, with intermediate clearance rates, delivered the highest absorbed dose to viable tumor cells. Antibody affinity and avidity facilitated the prolonged retention in radiosensitive areas of tumor, where most of the dose was deposited. The effectiveness of therapy could be enhanced further by matching the radionuclide with the antibody and tumor size. CONCLUSIONS: The model presented in this article allows the interaction between important radiobiologic parameters to be assessed and provides a tool for optimizing therapy in animal models and in patients. PMID- 11877754 TI - Volume reduction versus radiation dose for tumors in previously untreated lymphoma patients who received iodine-131 tositumomab therapy. Conjugate views compared with a hybrid method. AB - BACKGROUND: A Phase II study of previously untreated patients with malignant low grade follicular lymphoma given a combination of unlabeled tositumomab and tositumomab labeled with iodine-131 has recently been completed. The responses of these patients have been characterized, and for some of them tumor dosimetry during therapy has been estimated not only by pretherapy tracer conjugate views but also by a hybrid method. METHODS: Available patients were studied if they had had a pelvic or abdominal tumor evaluation by single photon emission computed tomography (SPECT) and achieved a partial response. A tumor outlined on the iodine-131 conjugate-view images was called a composite tumor. Its volume estimate came from multiple, not necessarily contiguous, regions of interest (ROI) on the pretherapy computed tomography (CT) scan. Its radiation dose was estimated from the weeklong series of pretherapy images and standard Medical Internal Radiation Dose methods. Computed tomography ROI were also grouped into smaller, contiguous volumes that defined individual tumors. Their radiation doses were estimated by the hybrid method. This method employed the activity measured for each individual tumor by a single intratherapy SPECT scan, as well as the tumor's volume, to individually normalize the composite time-activity curve as appropriate. The individual normalization factors then converted the composite radiation dose to radiation doses for individual tumors. Reduction in tumor volume was calculated for both composite and individual tumors at 12 weeks posttherapy. RESULTS: For 14 composite tumors in 10 patients, the median pretherapy volume was 170 cm(3). Application of a sigmoidal curve function to the plot of volume reduction versus radiation absorbed dose resulted in degeneration of the curve into a straight line with a negative slope. There was no statistical significance in the relationship (P = 0.73). For 43 individual tumors, the median pretherapy tumor volume was 26 cm(3). The plot of volume reduction versus dose was fairly well fit by a sigmoidal curve, and the relationship approached statistical significance (P = 0.06). The representation assigned 56% of the shrinkage to the effects of unlabeled tositumomab. For the subset of individual tumors with a pretherapy volume less than 10 cm(3) from 6 patients (n = 15), the relationship was significant (P = 0.03). The sigmoidal representation assigned only 12% of the shrinkage to unlabeled tositumomab, as contrasted with 72% for tumors with pretherapy volume greater than 10 cm(3). CONCLUSIONS: For patients who attained a partial response, analysis of individual tumors by a hybrid dosimetric method led to a dependence between volume reduction at 12 weeks and radiation dose that tended to be significant. The same was not true with dosimetry of composite tumors based on pretherapy conjugate views alone. It appeared that volume reductions from both unlabeled antibody and radiation dose were important in tositumomab therapy of lymphoma patients, with unlabeled antibody relatively more important for larger tumors. PMID- 11877755 TI - Model prediction of treatment planning for dose-fractionated radioimmunotherapy. AB - BACKGROUND: Clinical trials of radioimmunotherapy (RIT) often use dose fractionation to reduce marrow toxicity. The dosing scheme can be optimized if marrow and tumor cell kinetics following radiation exposure are known. METHODS: A mathematic model of tumor clonogenic cell kinetics was combined with a previously reported marrow cell kinetics model that included marrow stromal cells, progenitor cells, megakaryocytes, and platelets. Reported values for murine tumor and marrow cellular turnover rates and radiosensitivity were used in the model calculation. RESULTS: Given a tolerated level of thrombocytopenia, there is a fractionation scheme in which total radioactive dose administration can be maximized. Isoeffect doses that had different numbers of fractions and total radioactivity, but induced identical platelet nadirs of 20%, were determined. Assuming identical tumor uptake for all dose fractions, six tumor types were examined: early-responding tumors, late-responding tumors, and tumors that lacked a late-responding effect, with either constant or accelerated doubling time. For most tumor types, better tumor control (tumor growth delay and nadir of survival fraction) was predicted for a dosing scheme in which total radioactive dose was maximized. For late-responding tumors with accelerated doubling time, tumor growth delay increased, but the nadir of survival fraction became shallower as the number of fractions increased. CONCLUSIONS: A mathematic model has been developed that allows prediction of the nadir and duration of thrombocytopenia as well as tumor clonogenic cell response to various RIT doses and fractionation schemes. Given a maximum tolerated level of thrombocytopenia, the model can be used to determine a dosing scheme for optimal tumor response. PMID- 11877756 TI - Single tumor cell uptake and dosimetry of technetium-99m Fab' or minute anti-CD22 in low-grade B-cell lymphoma. AB - BACKGROUND: A patient with follicular lymphoma was investigated with 0.5 mg Fab' or minute anti-CD22 labeled with 1100 MBq technetium-99m ((99m)Tc). A computed tomography scan performed a week later revealed regression. This unexpected response prompted an investigation of single cell dosimetry of low-energy electron emitters. METHODS: Another patient with low-grade, unclassifiable B-cell lymphoma with a low expression of CD22 was injected with (99m)Tc anti-CD22. Blood samples were drawn 30 minutes, 4 hours, and 24 hours after injection. Lymphoma cells (CD19+) and T cells (CD3+), which served as control cells, were separated using a flow cytometer. The radioactivity of the two cell populations was measured in an NaI(Tl) well-type detector. The mean uptake per cell and absorbed dose were calculated. The CD22 expression of the patient's cells and of a B-cell lymphoma cell line (Raji) were assessed by flow cytometry for the extrapolation of the absorbed dose from the patient's cells to a cell line with higher CD22 expression. RESULTS: The average number of (99m)Tc atoms per CD19+ and CD3+ cell 4 hours postinjection were 5.4 and 0.054, respectively. Depending on the assumed ratio between antibody and CD22 molecules (1:2 or 1:1), the CD22 expression on the patient's cells and Raji cells varied from 2800 to 5700 and from 37,000 to 74,000 per cell, respectively. The average absorbed dose per cell ranged from 4 x 10(-7) to 0.1 grays (Gy). CONCLUSIONS: It seems feasible to assess the mean single tumor cell uptake of (99m)Tc targeted by Fab' or minute anti-CD22 in a patient's lymphoma using sorted cell populations, thereby allowing single cell dosimetry. Extrapolation of the absorbed dose from (99m)Tc to cells with higher CD22 expression was made and under certain conditions absorbed doses of 0.1 Gy were obtained, indicating the potential relevance of low-energy electron emitters to therapy. PMID- 11877757 TI - Choosing an optimal radioimmunotherapy dose for clinical response. AB - Clinical trials have documented the single-agent efficacy of radioimmunotherapy (RIT) in lymphoma, and several combination therapy studies are now in progress. RIT agents are currently becoming generally available for clinical use in lymphoma therapy. Solid tumors, which are notoriously less responsive to any single agent, have demonstrated clinically useful responses, albeit temporary, and multimodality studies have been instituted. However, a sincere debate continues regarding the basic parameters to be used to define appropriate therapeutic dosing when using this modality in clinical cancer care. It is a good time to reevaluate relevant dose response information from preclinical and clinical RIT. Preclinical studies have demonstrated abundant evidence of dose response in tumor and normal tissue in homogenous model systems; however, substantive variation occurs between the dose responses of tumors with low and variable (or shed) antigen expression, as well as between histologically different tumor models. Clinical studies of various heavily pretreated patient populations given several very different RIT pharmaceuticals have led to disparate conclusions regarding patient dosing methods and dosimetric predictions of toxicity and efficacy. Single-study data on previously untreated lymphoma patients with similar histology has demonstrated a correlation of imaging dosimetry with toxicity and tumor response. High-dose therapy with bone marrow support has also demonstrated a high tumor response rate and nonmarrow normal organ toxicities that correlate with the calculated dose to those organs from imaging. In iodine-131 ((131)I)--anti-CD20 studies, (131)I was demonstrated to have variable excretion, and estimated total-body radiation dose from tracer study proved a predictive surrogate for marrow toxicity. Yttrium-90 ((90)Y)--anti CD20, which has little (90)Y excretion from the body, demonstrated the injected dose per body weight to be more predictive of marrow toxicity than indium-111 ((111)In) tracer dosimetry methods in heavily pretreated patients, and showed maximal safety with standard mCi/kg therapy dosing. Variations in clinical RIT choices, dosing methods, and dosimetry methods emphasize the need to review the relevant information to date. Future clinical trial designs, the sophistication of dosimetry, treatment planning, and clinical treatment decisions should all be focused on achieving the best benefit-risk relationship for each patient. PMID- 11877758 TI - Application of extracorporeal immunoadsorption to reduce circulating blood radioactivity after intraperitoneal administration of indium-111-HMFG1-biotin. AB - BACKGROUND: Extracorporeal immunoadsorption (ECAT) is a method of reducing activity in radiosensitive organs by removing excess monoclonal antibodies (MAbs) from the blood. Previously, the authors experimentally evaluated ECAT based on the avidin-biotin concept after intravenous administration of radioimmunoconjugates. The aim of the current study was to determine whether ECAT could be used to reduce activity after intraperitoneal (i.p.) administration of indium-111((111)In)-HMFG1-biotin in rats, and to compare the pharmacokinetics of (111)In-HMFG1 with or without attached biotin after i.p. injection. METHODS: HMFG1, a murine immunoglobulin G(1) MAb that recognizes an epitope on the polymorphic epithelial mucin (PEM) antigen, was labeled with (111)In and then biotinylated. ECAT was explored from unseparated blood using an avidin-agarose adsorption column. Thirty rats were used as controls and 13 underwent ECAT. The whole-body (WB), blood, and organ activity were monitored. RESULTS: The binding capacity of (111)In-HMFG1-biotin to avidin was high. Biotinylation did not enhance the excretion of HMFG1. When ECAT was employed, the WB and blood radioactivity were reduced by 35-40% (P < 0.05) and 75--86% (P < 0.01), respectively. After the completion of ECAT, the activity uptake in organs was significantly decreased. CONCLUSIONS: ECAT was successfully applied after i.p. injection of the (111)In-HMFG-biotin MAb to reduce the radioactivity in the WB, blood, and radiosensitive organs. Due to redistribution of the radiolabeled MAbs during and after the completion of ECAT, the adsorption may have been prolonged or repeated. Biotinylation did not significantly change the biodistribution of the (111)In-HMFG1 in rats after intraperitoneal injection. PMID- 11877759 TI - A novel somatostatin conjugate with a high affinity to all five somatostatin receptor subtypes. AB - BACKGROUND: Somatostatin receptors (SRS, five subtypes) are expressed in a variety of human tumors, including most tumors of neuroendocrine origin, breast tumors, certain brain tumors, renal cell tumors, lymphomas, and prostate cancer. Somatostatin (SMS) triggers cytostatic and cytotoxic effects and has a general inhibitory effect on secretion mediated through its interaction with SRS. That is the basis for its use in the treatment of SRS-positive tumors. Radiolabeled SMS analogs can also be used for systemic radiotherapy and for diagnostic investigations. METHODS: Sms-14 was conjugated to a periodate-activated dextran70 (mean molecular weight, 70 kD) by reductive amination. The human tumor cell line LCC-18, from a neuroendocrine colonic tumor, was used for stable transfection with each SRS gene separately; transfection was achieved with the expression system TETon (Clontech, Palo Alto, CA). Clones were selected by culturing with G418 and hygromycin B, and positive clones were identified by reverse transcriptase-polymerase chain reaction and binding of iodine-125-labeled SMS-14. The binding affinity for each SRS subtype was then determined for the SMS-dextran conjugate (with SMS-14 used as a positive control). RESULTS: Sms-dextran70 showed high affinity binding to all five receptor subtypes. The IC50 values were between 3 and 80 nM. CONCLUSIONS: This conjugate has a long circulation half-life (i.e., approximately 27 hours after subcutaneous administration in mice) and, with high SRS pan-affinity demonstrated in this study, it has potential in the therapy of SRS-positive tumors. Currently, the clinical significance of SMS-dextran70 is being explored in a clinical Phase I-II study of patients with hormone-refractory prostate cancer. The outcome of this study will be reported when it is available. PMID- 11877760 TI - Targeted radiotherapy with [(90)Y]-SMT 487 in mice bearing human nonsmall cell lung tumor xenografts induced to express human somatostatin receptor subtype 2 with an adenoviral vector. AB - BACKGROUND: Novel approaches to increasing the therapeutic efficacy of targeted radiotherapy of cancer are required. One strategy to achieve this goal is to induce high-level expression of a receptor on the surface of tumor cells that can be targeted with a radiolabeled peptide. The objectives of this study were to 1) induce somatostatin receptor (SSTr2) expression in tumor cells using an adenovirus encoding the SSTr2 gene (AdSSTr2), 2) demonstrate tumor localization of [(111)In]-DTPA-D-Phe(1)-octreotide in AdSSTr2-injected tumors, and 3) show therapeutic efficacy with [(90)Y]-DOTA-D-Phe(1)-Tyr(3)-octreotide ([(90)Y]-SMT 487). METHODS: SSTr2 expression was validated in vitro by the binding and subsequent internalization of [(111)In]-DTPA-D-Phe(1)-octreotide (21.3% per mg of total protein) in A-427 cells infected with AdSSTr2. In vivo imaging confirmed 5- to 10-fold greater uptake 5.5 hours after intravenous administration of [(111)In] DTPA-D-Phe(1)-octreotide in AdSSTr2-injected tumors relative to control tumors. For therapy studies, mice bearing established subcutaneous A-427 tumors were given two intratumoral injections of AdSSTr2 1 week apart, followed by an intravenous injection of 400 microCi or 500 microCi [(90)Y]-SMT 487 at 2 and 4 days after each adenoviral administration. Control animals either were not treated or were administered 500 microCi [(90)Y]-SMT 487 with no AdSSTr2 injection. RESULTS: These studies showed that untreated animals and animals treated with no virus and 500 microCi [(90)Y]-SMT 487 had median tumor quadrupling times of 16 and 25 days, respectively. Mice administered AdSSTr2 and either 400 microCi or 500 microCi of [(90)Y]-SMT 487 demonstrated significantly longer median tumor quadrupling times of 40 and 44 days, respectively (P < 0.02). CONCLUSIONS: These studies are the first to demonstrate in vivo therapeutic efficacy using a radiolabeled peptide targeted to a receptor expressed on the surface of tumor cells following gene transfer. Future studies will focus on the optimization of this approach. PMID- 11877761 TI - Idiotypic-anti-idiotypic complexes and their in vivo metabolism. AB - BACKGROUND: Different strategies can be used to improve the tumor:non-tumor ratio of radiolabeled antibodies in immunotargeting. One approach is to use secondary antibodies to clear out redundant, circulating primary antibodies. In the current study, the in vitro complex formation and in vivo clearing capabilities and metabolism of the monoclonal antibody TS1 and its monoclonal anti-idiotype, alphaTS1, were studied. METHODS: Complex formation studies were performed using polyacrylamide gel electrophoresis (PAGE), gel permeation chromatography, and electron microscopy. The clearance and metabolism of the complexes were studied in nude mice. RESULTS: PAGE and gel permeation chromatography showed that more than 70% of the antibodies formed complexes. The electron microscopy studies revealed that the complexes formed between TS1 and alphaTS1 are mainly ring shaped (66.6-73.4%), comprising 4 to > 8 antibodies. These rings consist of equal numbers of idiotype and anti-idiotype. The most commonly observed complexes were tetrameric rings (26.8-40.5%), hexameric rings (10.7-11.9%), and rings containing more than eight monoclonal antibodies (6.6-14-4%). The in vivo study illustrated that within 24 hours 80% of the total nuclide content had been degraded and excreted via the urine, compared with 25% for similarly treated mice that did not receive any anti-idiotype. CONCLUSIONS: Interestingly, the electron microscopy study demonstrated that dimers were rare (0.4-1.2%), probably reflecting a location of epitopes incompatible with tight, sterically constrained dimeric interactions; insufficient flexibility of the immunoglobulin G1 subtype hinge regions; or both. The anti-idiotypic clearing mechanisms proved efficient in nude mice. In vivo metabolic studies indicate that the accumulation and degradation of TS1/alphaTS1 immune complexes, to a large extent, take place in the liver, where a substantial amount was detected as soon as 1 hour after anti-idiotype injection. PMID- 11877762 TI - The combination of external beam radiotherapy and experimental radioimmunotargeting with a monoclonal anticytokeratin antibody. AB - BACKGROUND: Doses to tumors of up to 80 grays (Gy) have been postulated to eradicate solid experimental tumors with radiommunotargeting, but this value has proved difficult to reach. Combining two treatment modalities, external beam radiotherapy and radioimmunotargeting, could potentially give rise to a number of advantages. METHODS: The purpose of this study was to detect potential benefits with different treatment timing strategies when combining external beam radiotherapy and radioimmunotargeting, with the anticytokeratin monoclonal antibody (MAb) TS1 injected into a nude mouse model carrying subcutaneous human HeLa Hep 2-cell tumors. Cytokeratins are present in necrotic regions within tumors, thereby providing a potential increase in binding sites for TS1 if combined with external beam radiotherapy. External beam radiotherapy was given before, after, and simultaneously with injection of radiolabeled MAb. RESULTS: The highest yields in terms of total accumulated dose (Gy), percentage of injected activity per gram of tumor tissue, and accumulated dose per injected activity (Gy/MBq) were seen in the group receiving external beam radiotherapy prior to MAb-injection. CONCLUSIONS: Enhanced effects may be achievable by combining external beam radiotherapy with experimental radioimmunotargeting using the monoclonal anticytokeratin antibody TS1, if the radiotherapy is given prior to MAb injection. PMID- 11877764 TI - Rationales, evidence, and design considerations for fractionated radioimmunotherapy. AB - Although fractionation can be used in a discrete radiobiologic sense, herein it is generally used in the broader context of administration of multiple, rather than single, doses of radionuclide for radioimmunotherapy (RIT) or other targeted radionuclide therapies. Fractionation is a strategy for overcoming heterogeneity of monoclonal antibody (MAb) distribution in the tumor and the consequent nonuniformity of tumor radiation doses. Additional advantages of fractionated RIT are the ability to 1) provide patient-specific radionuclide and radiation dosing, 2) control toxicity by titration of the individual patient, 3) reduce toxicity, 4) increase the maximum tolerated dose (MTD) for many patients, 5) increase tumor radiation dose and efficacy, and 6) prolong tumor response by permitting treatment over time. However, fractionated RIT has logistic and economic implications. Preclinical and clinical data substantiate the advantages of fractionated RIT, although the radiobiology for conventional external beam radiotherapy does not provide a straightforward rationale for RIT unless fractionation leads to more uniform distribution of radiation dose throughout the tumor. Preclinical data have shown that toxicity and mortality can be reduced while efficacy is increased, thereby providing inferential evidence of greater uniformity of radiation dose. Direct evidence of superior dosimetry and tumor activity distribution has also been found. Clinical data have shown that toxicity can be better controlled and reduced and the MTD extended for many patients. It is clear that fractionated RIT can only fulfill its potential if the effects of critical issues, such as the number and amount of radionuclide doses, the radionuclide physical and effective half-life, and the dose interval, are better characterized. PMID- 11877763 TI - Combined modality radioimmunotherapy. Promise and peril. AB - BACKGROUND: Single-agent radioimmunotherapy (RIT), although potentially useful for slowing solid tumor growth, has not been effective in curing aggressive tumors, such as breast cancer. These cancers typically have p53 mutations and are less susceptible to apoptosis, the apparent mechanism of cell death from low dose rate radiation. Thus, synergistic or combined modality radioimmunotherapy (CMRIT) agents are needed to increase radiosensitivity for therapeutic enhancement without additive toxicity. METHODS: To assess synergy in CMRIT in a breast cancer xenograft model, we evaluated RGD peptide EMD 121974, an inhibitor of alpha(v)beta(3) integrin; paclitaxel, an antimicrotubule; IMC-C225, a monoclonal antibody to epidermal growth factor receptor (EGFR); and bcl-2 antisense oligonucleotide G3139. Groups of mice received (90)Y-DOTA-ChL6 in combination with each agent. Tumor size, survival, and blood counts were monitored for efficacy and toxicity. Immunopathologic evaluation of apoptosis was performed at selected time points after RIT and RIT + RGD CMRIT. RESULTS: CMRIT with RGD peptide increased apoptosis and resulted in 57% cures, compared with 0% cures with RIT alone. CMRIT with paclitaxel after RIT increased cures to 88%, compared with 25% cures with RIT before paclitaxel administration. CMRIT with IMC-C225 resulted in up to 20% cures if given before RIT. A time-dependent increase in toxicity was observed with IMC-C225 after RIT. CMRIT with bcl-2 antisense G3139 resulted in no cures and an increased rate of regrowth compared with RIT alone. CONCLUSIONS: Some combined modality therapies resulted in higher numbers of cures, while others decreased cures and responses and increased toxicity compared with RIT alone. These results support the potential for CMRIT but illustrate the complexity of predicting the efficacy and toxicity and the importance of the relationship between dose and sequence of administration. PMID- 11877765 TI - Radiation dosimetry results for Zevalin radioimmunotherapy of rituximab refractory non-Hodgkin lymphoma. AB - BACKGROUND: Zevalin consists of a murine anti-CD20 monoclonal antibody (ibritumomab) conjugated to the linker-chelator tiuxetan, which securely chelates indium-111 ((111)In) for imaging and dosimetry and yttrium-90 ((90)Y) for radioimmunotherapy (RIT). Previous trials involving rituximab-naive patients have demonstrated excellent targeting of Zevalin to CD20+ B-cell non-Hodgkin lymphoma with minimal uptake in normal organs. The purpose of this trial was to perform (111)In-Zevalin imaging in patients with rituximab-refractory tumors to determine normal organ dosimetry. METHODS: Twenty-seven patients were given an imaging dose of 5 mCi (185 MBq) (111)In-Zevalin on Day 0, evaluated with dosimetry, and then given a therapeutic dose of 0.4 mCi/kg (15 MBq/kg) (90)Y-Zevalin on Day 7. Both Zevalin doses were preceded by an infusion of 250 mg/m(2) rituximab to clear peripheral B cells and improve Zevalin biodistribution. Residence times for (90)Y in blood and major organs were estimated from (111)In biodistribution, and the MIRDOSE3 computer software program was used to calculate absorbed radiation doses to organs and red marrow. RESULTS: Median estimated absorbed radiation doses from (90)Y-Zevalin were 8.1 Gray (Gy) (range, 4.2-23.0 Gy) to the spleen, 5.1 Gy (range, 2.6-12.0 Gy) to the liver, 2.0 Gy (range, 1.4-5.3 Gy) to the lungs, 0.22 Gy (range, < 0.01-0.66 Gy) to the kidneys, and 0.74 Gy (range, 0.29-1.2 Gy) to the red marrow. These results are consistent with those from earlier Zevalin trials in rituximab-naive patients. Hematologic toxicity was manageable and did not correlate with estimates of red marrow or total-body absorbed radiation dose. CONCLUSIONS: Zevalin treatment of rituximab-refractory follicular NHL patients at 0.4 mCi/kg resulted in acceptable estimates of absorbed radiation dose to organs, similar to those observed in other Zevalin-treated populations. PMID- 11877766 TI - Outpatient radioimmunotherapy with Bexxar. Closed, clean air reservoir minimizes personnel radiation exposure. AB - BACKGROUND: Radioimmunotherapy (RIT) with Bexxar (tositumomab and iodine-131 tositumomab; Coulter Pharmaceutical, South San Francisco, CA) has been shown to be effective in the treatment of low-grade and transformed low-grade non-Hodgkin lymphoma (NHL). METHODS: Patient-specific dosimetry with 5 mCi of iodine-131 tositumomab preceded by 450 mg of tositumomab was utilized to calculate the radionuclide dose needed to deliver 75 cGy whole-body radiation (65 cGy for platelet counts of 100,000-149,000/mm(3)). To safely infuse the approximately 95 mCi (range, 52-211mCi) of iodine-131 needed for this treatment, a shielded, closed system was developed to minimize radiation exposure for personnel administering the treatment infusions and to eliminate possible release of aerosolized iodine-131. RESULTS: Twenty-five patients who could be evaluated were infused with a single course of iodine-131 tositumomab therapy and achieved a 76% total response rate at 3 months (32% complete response [CR], 44% partial response [PR]); 59% total response at 6 months (40% CR, 18% PR); and 38% total response at 12 months (31% CR, 6% PR). Administration of RIT using our unique, totally closed system significantly reduced personnel exposure and potential for radioactive spills. The sum of all individuals who administered and monitored the infusions was < 120 mRem whole body exposure over 22 months, well within the ALARA (as low as reasonably achievable) Level I guideline limits. No radioiodide was detectable in the thyroid of any staff member. CONCLUSIONS: In NHL patients who had experienced failure with conventional therapy, RIT with iodine-131 tositumomab therapy was safe and effective. Response rates obtained were equivalent to those obtained at the university medical centers where the Phase I-III clinical trials were performed. PMID- 11877767 TI - High-dose myeloablative radioimmunotherapy of mantle cell non-Hodgkin lymphoma with the iodine-131-labeled chimeric anti-CD20 antibody C2B8 and autologous stem cell support. Results of a pilot study. AB - BACKGROUND: CD20 has been used successfully as a target molecule for conventional low-dose, as well as high-dose, myeloablative radioimmunotherapy (RIT) of B-cell non-Hodgkin lymphoma (NHL). Mantle cell lymphoma (MCL) is an especially aggressive, prognostically unfavorable subtype of B-cell NHL, associated with an overall 5-year survival rate of less than 20%. Recent evidence has failed to show convincing therapeutic efficacy of conventional, nonmyeloablative RIT in patients with MCL. The aim of this pilot study was to investigate whether high-dose, myeloablative RIT with the iodine-131 ((131)I)-labeled chimeric anti-CD20 antibody C2B8 (rituximab, obtained as Mabthera from Roche Pharma, Reinach, Switzerland) is therapeutically effective in MCL patients. METHODS: A total of seven patients with chemorefractory or relapsed MCL were studied in this pilot trial. All had relapsed after high-dose chemotherapy with autologous stem cell transplantation (four of them combined with 12 grays (Gy) total-body irradiation). A diagnostic-dosimetric study was performed with approximately 10 mCi of (131)I-labeled C2B8 at a protein dose of 2.5 mg per kg body weight, in order to assess its biodistribution and dosimetry. If splenic pooling was observed, as is typically the case in patients with splenomegaly, the protein dose was doubled in additional studies until a "favorable" biodistribution was obtained. Therapy was performed with myeloablative doses of 261-495 mCi of (131)I labeled C2B8 at the previously optimized protein dose, aiming at lung doses less than-or-equal 27 Gy. Homologous stem cell support was provided. Clinical follow up was obtained at 3-month intervals for up to 38 months (median observation time, 25 months). Overall, in six patients the 2.5 mg/kg protein dose was used, whereas in one patient with splenomegaly, 10 mg/kg was necessary to overcome the splenic antigenic sink. RESULTS: Blood cell nadirs were reached at 2-3 weeks after therapy infusion, but all patients reengrafted at 7-10 days after stem cell reinfusion. Nonhematologic toxicity was restricted to mild-to-moderate nausea, fever, transient bilirubin, or liver enzyme elevations. One patient with preexisting alcoholic cirrhosis experienced a deterioration of liver function. Despite thyroid blocking, 5 of 7 patients developed hypothyroidism, requiring thyroxine substitution at 6-18 months after RIT. Six patients experienced a complete and one a good partial remission. Five patients were still in CR at the time this article was written, and six are still alive for more than 3 years; one patient relapsed locally at 3 months and one systemically at 26 months after RIT. CONCLUSIONS: High-dose myeloablative radioimmunotherapy with (131)I-labeled anti CD20 antibodies seems to be associated with a high response rate and moderate toxicity in patients with MCL. Further follow-up to monitor the long-term outcome as well as systematic prospective clinical studies are indicated. PMID- 11877769 TI - Hyperglycemia and congenital malformations in insulin-dependent diabetes mellitus: a brief summary and evaluation. PMID- 11877768 TI - Radioimmunotherapy of small-volume disease of metastatic colorectal cancer. AB - BACKGROUND: Whereas radioimmunotherapy (RIT) has shown disappointing results in bulky, solid tumors, preclinical results in small-volume disease and in an adjuvant setting are promising. In a previous Phase I study, the authors had encouraging results with the iodine-131 ((131)I)-labeled humanized anti carcinoembryonic antigen (anti-CEA) antibody (MAb) hMN-14 in small-volume disease of colorectal cancer. The aim of this study was to evaluate, in a subsequent Phase II trial, the therapeutic efficacy of this (131)I-labeled humanized anti CEA antibody in colorectal cancer patients with small-volume disease or in an adjuvant setting. METHODS: Thirty colorectal cancer patients, with small-volume metastatic disease (n = 21; all lesions less-than-or-equal 3.0 cm, and chemorefractory to 5-fluorouracil and folinic acid) or in an adjuvant setting (n = 9), 4-6 weeks after surgical resection of liver metastases with curative intention, were studied. The patients were given a single injection of (131)I-hMN 14 immunoglobulin G at a 60 mCi/m(2) dose level, which was shown to be the maximum tolerated dose in the previous Phase I study. Follow-up was obtained at 3 month intervals for as long as 36 months. RESULTS: At a mean blood-based red marrow dose of 1.8 +/- 0.8 Gy, myelotoxicity was the only toxicity observed, but only 1 of 28 assessable patients developed transient Grade 4 thrombocytopenia. Of the 21 patients with radiologically documented lesions, 19 were assessable. Three experienced partial remission and eight showed minor responses up to 15 months in duration (corresponding to an objective response rate of 16% and an overall response rate of 58%; the mean duration of response was 9 months). At the time this article was written, seven of nine patients in the adjuvant setting had remained free of disease for up to 36 months (one patient relapsed after 6 months and another after 30 months), whereas the relapse rate in a historical control group receiving chemotherapy was 67% over the same time period. Five patients with radiologically documented lesions, having experienced at least disease stabilization as a consequence of RIT, were retreated at the same 60-mCi/m(2) dose level at 8-16 months after the first therapy. No evidence of increased toxicity was observed (no hematologic toxicity was higher than Grade 3). Two of four assessable retreated patients experienced partial remissions; one of these four again experienced disease stabilization as a consequence of the second radioantibody therapy injection. CONCLUSIONS: These data suggest that RIT is a safe and effective form of therapy for small-volume colorectal cancer and has potential as treatment for colorectal cancer in an adjuvant setting. Toxicity is restricted to mild and transient leuko- and thrombocytopenia. Retreatment seems to be a feasible option. A prospective randomized comparison with standard chemotherapy is indicated. PMID- 11877771 TI - Do platelet-released phospholipids play a role in cardiovascular malformations and heritable coangulopathies? PMID- 11877773 TI - Infants with single ventricle: a population-based epidemiological study. AB - BACKGROUND: Single ventricle, a rare congenital cardiac defect, often occurs as part of a complex group of cardiovascular abnormalities. Little is known of its epidemiologic associations. METHODS: Using data from the Baltimore-Washington Infant Study [BWIS], (1981-89), a population based case-control study of cardiovascular malformations, infants with single ventricle were evaluated with respect to infant and family characteristics and maternal and paternal exposures. The cases were analyzed according to presence/absence of abnormal cardio-visceral situs. Controls were 3,572 infants without heart defects randomly selected from the regional cohort of live births. Odds ratios and 95% confidence intervals were used as measures of association. RESULTS: Single ventricle occurred in 1.25% of infants with congenital cardiovascular defects in the BWIS. Fifty-five infants had single ventricle. In 48 families (87.3%) the parents were interviewed. Thirty three infants had normal situs and 15 had abnormal situs. Paternal alcohol consumption (OR = 2.0, 95% CI 1.1-3.9) and paternal cigarette smoking (OR = 2.4, 95% CI 1.1-5.1) were associated with all cases of single ventricle. These associations were even stronger in the subset of infants with abnormal situs. Maternal history of a previous induced abortion was also associated with infants born with abnormal situs (OR = 3.2, 95% CI 1.1-11.5). Paternal marijuana use was associated with cases of single ventricle in normal situs (OR = 2.2, 95% CI 1.0 5.2). CONCLUSIONS: Potential risk factors included paternal smoking and alcohol consumption, highlighting the need for future studies to consider environmental factors in the pathogenesis of this cardiac defect. PMID- 11877774 TI - Altered mandibular development precedes the time of palate closure in mice homozygous for disproportionate micromelia: an oral clefting model supporting the Pierre-Robin sequence. AB - BACKGROUND: Development of the human craniofacial anatomy involves a number of interrelated, genetically controlled components. The complexity of the interactions between these components suggests that interference with the spaciotemporal interaction of the expanding tongue and elongating Meckel's cartilage correlates with the appearance of cleft palate. Mice homozygous for the semi-dominant Col2a1 mutation Disproportionate micromelia (Dmm), presenting at birth with both cleft palate and micrognathia, provide the opportunity to test the hypothesis that mandibular growth retardation coincides with formation of the secondary palate as predicted from our understanding of the Pierre Robin sequence. The present study was conducted in embryonic day 14 (E14) mice, 1 day before palate closure, to describe the relationship between growth of the lower jaw/tongue complex versus genotype of the embryo. METHODS: Whole heads, isolated from E14.25, E14.5 and E14.75 wild-type and homozygous mutant embryos, were fixed in Bouin's solution, embedded in paraffin, and serially sectioned. Mid-sagittal sections, stained with toluidine blue, were used to estimate growth of both tongue and lower jaw (Meckel's cartilage length) during a 12-hr period preceding palate closure. RESULTS: In control embryos, the largest increase in Meckel's cartilage length occurred between E14.5 and E14.75. Compared to control, the mean Meckel's cartilage length in the mutant was similar at E14.25, but was significantly less at E14.5 and E14.75. Absolute tongue size in control embryos increased linearly during this period of E14.25 to E14.75. Relative to the rapidly growing Meckel's cartilage, however, relative tongue size in control embryos actually decreased over time. Absolute tongue size in the mutant was not significantly different from that of control at any of the embryonic stages examined, however, relative tongue size in the mutant was significantly greater at E14.75 compared to control. CONCLUSION: Mandibular growth retardation, coupled with relative macroglossia in E14 Dmm/Dmm mice, suggests that the concerted development of the palate and lower jaw complex in the mutant is aberrant. Detection of micrognathia and pseudomacroglossia in homozygotes, before the time of palate closure, supports the hypothesis that a relationship exists between growth retardation of Meckel's cartilage and malformation of the secondary palate, as predicted by the Pierre-Robin sequence. PMID- 11877775 TI - Congenital malformations in offspring of Vietnamese women in California, 1985-97. AB - BACKGROUND: Little is known about reproductive outcome risks for Vietnamese women delivering infants and fetuses in the U.S. METHODS: Using data from a large population-based registry, we explored risks of selected congenital malformation phenotypes in offspring of Vietnamese women in California. Data were derived from the California Birth Defects Monitoring Program, a population-based active surveillance system for collecting information on infants and fetuses with congenital malformations using multiple source ascertainment. Approximately 3.4 million births (liveborn and stillborn) occurred during the ascertainment period, 1985-97. Information on maternal race/ethnic background was obtained from California birth certificate and fetal death files. Vietnamese women delivered 45,453 births and 1,257,853 births were delivered to non-Hispanic white women. RESULTS: The overall prevalence of structural congenital malformations was 1.92 among Vietnamese and 2.63 among non-Hispanic whites per 100 births and fetal deaths. Grouping by 20 3-digit malformation codes of the International Classification of Diseases-Ninth Revision revealed relative risks of 0.8 or less for spina bifida, eye, upper alimentary, genital, urinary, musculoskeletal, "other" limb, and "other" musculoskeletal anomalies, and relative risks of 1.3 or more for anencephaly and chromosomal anomalies. Grouping by the more specific 4 digit malformation codes revealed 50, among 178, malformation groupings with associated relative risks of >or=1.3 or or=3 minor anomalies (11.7% vs. 7.6%, RR 1.53; 95% CI 0.61, 3.82). Furthermore, no pattern of malformation was identified. There were no significant differences between the groups in spontaneous pregnancy loss (6.1% vs. 8.2%, P = 0.65) or premature delivery (8.6% vs. 7.7%, P = 0.95). Birth weight and head circumference were significantly increased in the exposed group; however, these differences were not associated with anorexiant use itself. The rate of gestational diabetes was significantly increased in pregnant women who took phentermine/fenfluramine during the first trimester of pregnancy. CONCLUSIONS: Although it is not possible from this study to rule out weak to moderate associations, the lack of an increased risk of spontaneous pregnancy loss, and major or minor anomalies in the offspring of women who took phentermine/fenfluramine at the recommended daily dose during the first trimester of pregnancy is reassuring. PMID- 11877777 TI - Mitochondrial transduction of ocular teratogenesis during methylmercury exposure. AB - BACKGROUND: The purpose of the present study was to investigate the correlation between MeHg developmental toxicity and mitochondrial 16S ribosomal RNA (16S rRNA) expression in the embryonic forebrain and pharmacological intervention with PK11195, a ligand for the mitochondrial peripheral-type benzodiazepine receptor (Bzrp). METHODS: Pregnant CD-1 mice were dosed with methylmercury (II) chloride (MeHg) with or without 4 mg/kg PK11195 on Day 9 of gestation. Fetuses were examined on Day 9 (RT-PCR), Day 15 (histology), and Day 17 (teratology). RESULTS: MeHg (10 mg/kg) induced microcephaly, microphthalmia and cleft palate. The mean incidences of malformed fetuses were 47.7% with MeHg (P < 0.001) and 19.2% with PK11195 co-treatment (P < 0.01 for rescue). Cleft palates were 12.8% and 1.5%, respectively. An estimate of neurocranial circumference revealed a small (5%) but highly significant (P < 0.001) reduction that was rescued in a subset of co treated fetuses (P < 0.05). RT-PCR analysis of the Day 9 forebrain revealed inhibition of 16S rRNA expression 3.0 hr after 5 mg/kg MeHg exposure (P < 0.001). This effect was rescued with PK11195 (P < 0.001). Preliminary findings revealed a similar response-rescue in cultured embryos exposed to 1 microM Hg(II) when exogenous 5-aminolevulinic acid (ALA) was added. Protoporphyrin-IX (PP9), the penultimate precursor to heme and an endogenous ligand of the Bzrp, increased in a manner that was ALA-dependent and PK11195-sensitive. CONCLUSION: At least some teratological effects of Hg appear linked with late steps in the heme biosynthesis pathway through the Bzrp. PK11195, a ligand for these mitochondrial receptors, significantly lessens the risk of microphthalmia, microcephaly, and cleft palate in Hg-poisoned embryos. PMID- 11877783 TI - Danger of damaging the medial branches of the posterior rami of spinal nerves during a dorsomedian approach to the spine. AB - Postoperative atrophy of the deep back muscles may be caused by denervation during a dorsomedian approach to the thoracolumbar spine; ensuing instability of the spine with poor clinical results, perhaps due to such muscle loss, has been observed in 11.7% of cases (Sihvonen et al., 1993, Spine 18:575--581). More specifically, this complication may be caused by damaging the medial branches of the posterior rami of the spinal nerves during lateral retraction of the muscles. To investigate the anatomic topography of the medial branches of the posterior rami of the spinal nerves, 18 carbol-formol-fixed specimens were dissected using an operation microscope; also, 3 fresh cadavers were cut in horizontal and vertical planes with a rotary cryotome to confirm the anatomic topography observed in the fixed specimens. In the thoracolumbar spine the medial branch of the posterior ramus of the spinal nerve is subject to ligamentous fixation by the strong fibers of the mammillo-accessory ligament, which extends between the mammillary process and accessory process infero lateral to the superior articular process. When the dorsomedian approach to the thoracolumbar spine is enlarged laterally to the articular processes by retracting the paraspinous muscles, the medial branches of the posterior rami of the spinal nerves are endangered. This may cause postoperative pain as well as dynamic instability beyond the corresponding segments. The results of our anatomic study suggest that the posterior surgical midline approach to the thoracolumbar spine should not be enlarged laterally to the articular processes to prevent injury to the medial branches of the posterior rami of the spinal nerves. PMID- 11877784 TI - Lumbar intrathecal ligaments. AB - A meticulous examination was performed on 56 vertebral columns from cadavers between 64 and 89 years of age. Identification of all contents within the dural sac was completed; however, the main focus was the cauda equina and lumbar region. In addition to scope dissection, radiographs and histological preparations were used to identify structures, tissue types, and any possible pathology. Discrete intrathecal ligamentous bands were observed in all cadavers examined. They were found randomly binding the dorsal nerve roots of the cauda equina to the dura. Occasional binding of the ventral nerve roots to the dorsal roots was observed. Histological examination demonstrated a dense collagen ligament varying between 0.13 and 0.35 microm in thickness and from 3 mm to 3.5 cm in length. The average number of ligaments found per cadaver was 18. These ligaments displayed a broad base attachment to the nerve root or dura of approximately 3 mm. Looping of the nerve roots associated with these ligaments was seen in one cadaver with a burst fracture. Electron microscopic studies of these ligaments demonstrated similarities to denticulate ligaments. It is suggested that the intrathecal ligaments represent remnants from fetal development of the denticulate ligaments. PMID- 11877785 TI - Measuring movement at the low back. AB - The clinical importance of movements of the human spine is increasingly being realized but their measurement is difficult to carry out. Traditionally, measurement of spinal motion focuses on the lumbar spine as the mobile region and movement at the lower thoracic spine is largely ignored. The aim of this study was to test the hypothesis that sagittal movement at the low back is not confined to the lumbar spine. The range of sagittal movement and the amount of skin distraction during trunk flexion was studied in 10 normal subjects. Angular movement in the sagittal plane between the vertebra prominens and S2 ranged between 40 degree and 83 degree among the subjects. Concomitant skin distraction showed that a mean of 32% (range 20-41%) of the movement between the upper thorax and sacropelvis occurred cranial to T12. We therefore suggest that the term "low back" should be applied to a series of mobile vertebrae in the lower thoracic and lumbar spines and that motion of this region in the sagittal plane should be considered as the movement of an immobile upper thoracic spine relative to the sacropelvis. PMID- 11877787 TI - Morphometric analyses of the mandible in prepubertal craniofacial microsomia patients treated with an inverted-L osteotomy. AB - To analyze changes in mandibular form associated with an inverted-L osteotomy and autogenous bone graft, preoperative, early postoperative (EPO), and late postoperative (LPO) lateral cephalographs of 14 children (mean age approximately 9 years) with unilateral craniofacial microsomia (CFM) were scanned and nine mandibular landmarks digitized. Average mandibular geometries, scaled to an equivalent size, were generated using Procrustes superimposition. Cephalometry, Euclidean distance matrix analysis (EDMA), and thin-plate spline (TPS) analyses were carried out on mean mandibular configurations. Cephalometric results showed increases in oblique mandibular length (approximately 9% on average, P < 0.05) and increased ramus height (P < 0.05). Similarly, using EDMA there were also significant differences (P < 0.05) between the mean preoperative, EPO, and LPO configurations. The most demonstrable EPO change in the mandibular configuration using EDMA was increased oblique length (approximately 11%). This improvement depended on ramus oblique lengths increasing by approximately 26%, ramus height increasing by approximately 25%, and mandibular body length increasing by approximately 5%. For TPS analysis, affine and nonaffine changes contributed to the total spline. In all three comparisons the affine transformation showed an antero-inferior rotation of the mandibular configuration. For nonaffine changes the EPO configuration indicated a supero-inferior stretch of the mandibular configuration. The nonaffine LPO changes maintained the supero-inferior stretch of the mandibular configuration. It is concluded that improvements in the lateral facial profile of CFM patients can be achieved using an inverted-L osteotomy, with little relapse approximately 2 years postoperatively. PMID- 11877786 TI - Microsurgical anatomy of the facial nerve trunk. AB - Dissection and manipulation of the facial nerve (FN) trunk between its exit from the cranial base through the stylomastoid foramen (SMF) and its bifurcation is a critical step in various otologic, plastic and neurosurgical procedures. This study demonstrates the anatomical relationships and variability of the FN trunk with emphasis on some important morphometric data, particularly with relevance to hypoglossal-facial nerve anastomosis (HFA). Bilateral microsurgical dissection was performed on twenty-three human cadavers fixed with formalin. The whole trunk of the FN was exposed, its diameter at the SMF and its length were measured, its branches were observed and the site of its bifurcation was determined. Anastomotic connections with other nerves and blood supply of the trunk were studied. The FN invariably emerged from the cranial base through the SMF. Its diameter upon its emergence from the foramen was 2.66 +/- 0.55 mm. Two branches consistently originated from the trunk: the posterior auricular nerve and the nerve to the digastric muscle. Less consistent were the communicating branch with the glossopharyngeal nerve and the nerve to the stylohyoid muscle. The bifurcation of the FN occurred before its penetration into the parotid gland in 15% of cases and within the gland in 85%. The length of the FN trunk was 16.44 +/ 3.2 mm. Anastomoses between the FN and other nerves were observed in one-third of the dissections. The blood supply to the FN trunk was provided by the stylomastoid artery that was identified in 91% of cases. Understanding the microsurgical anatomy of the FN trunk is essential for performing any surgical procedure in the relevant region. Surgical implications of this study are presented with emphasis on HFA surgery. PMID- 11877788 TI - Posterior dislocation of the sternoclavicular joint: a case report and review of the clinical anatomy of the region. AB - Posterior dislocation of the sternoclavicular joint is an uncommon injury. Since 1824, when Sir Astley Cooper described the injury, little more than 100 cases have been described, and the majority of these have been in the last decade, identified by computed tomography. The significant morbidity and mortality associated with this injury is based upon serious damage to important anatomical structures found in the vicinity of the joint. We present a case report, describe the relevant clinical anatomy of this region, and review associated complications. PMID- 11877789 TI - Relative positions of the arteries and veins on the dorsolateral surface of the human cerebrum. AB - We investigated whether arteries pass superficial to veins or whether veins pass superficial to arteries at artery-vein crossings on the anterior, dorsolateral, and posterior surfaces of the human cerebrum. We examined a total of 2,266 artery vein crossings on 40 sides of 20 cadavers. At 2,059 crossings (91%), the vein passed superficial to the artery. Thus, vein (V), artery (A), and nerve (N), if we regard the cerebrum as nerve, were generally arranged in the order VAN from the superficial to the deep layers. This concept is important for a positional understanding of blood vessels on the cerebrum and it is useful for the understanding of fluid-drainage pathways from the cerebral cortex in various pathological conditions. PMID- 11877790 TI - Variability in human quadriceps muscles: quantitative study and review of clinical literature. AB - Knowledge of variations in normal human thigh musculature is important for the interpretation of various clinical and biomedical investigations. In this study, cross-sections from whole thighs of 34 cadavers were analyzed qualitatively and morphometrically. Sections were cut from right and left limbs at three levels in the region often used for muscle biopsy. Measurements were made of limb circumferences and cross-sectional areas of the three vasti and of the femur, and of the extent of overlap and fusion between the vasti on the lateral aspect of the thigh. Limb circumference proved to be a good predictor of total cross sectional area of the quadriceps muscle. Large individual variations were found for most measurements but especially for the amount of fusion between the muscle bellies of vastus lateralis and vastus intermedius. Significant differences were found for most variables between the three levels, but there were few differences between sides except for the cross-sectional area of vastus lateralis which was greater on the right at the most distal level. These variations in size and form of thigh musculature should be considered when interpreting data obtained from muscle biopsy, imaging, biomechanical, electromyographic, and functional studies of the human thigh. Review of the literature on variability of quadriceps muscle and its clinical relevance showed a diversity of approaches but insufficient definitive data suitable for interpreting functional outcomes in response to exercise or following surgery. Further work is suggested combining current anatomical datasets with advanced imaging techniques capable of determining muscle fiber orientation and fiber type composition, and including 3D reconstruction. PMID- 11877791 TI - Martin-Gruber anastomosis revisited. AB - Based on a study of 70 human cadavers (31 male, 39 female) and on cases described previously, we propose a new classification of the Martin-Gruber anastomosis, a neural connection between the median and ulnar nerves in the forearm. The anastomosis was found in 16 (22.9%) cadavers, being bilateral in three (18.7%) and unilateral in 13 (81.3%), five right and eight left. It occurred in eight (25.8%) of the 31 male cadavers and in eight (20.5%) of the 39 females. Therefore, the anastomosis was found in 19 (13.6%) of the 140 forearms. In Pattern I (89.5%) the anastomosis was made by only one branch, whereas in Pattern II (10.5%) it was made by two. The individual branches were classified as Types a, b, and c based on the nature of their origin from the median nerve. Type a (47.3%) arose from the branch to the superficial forearm flexor muscles, Type b (10.6%) from the common trunk, and Type c (31.6%) from the anterior interosseous nerve. Pattern II was a duplication of Type c (10.5%). The anastomotic branch took an oblique or arched course before joining the ulnar nerve, undivided in 15 cases, but divided into two branches in four cases. The anastomosis passed in front of the ulnar artery in four cases, behind it in six, and in nine cases it was related to the anterior ulnar recurrent artery. PMID- 11877792 TI - Intramuscular Martin-Gruber anastomosis. AB - The incidence and morphology of the intramuscular Martin-Gruber anastomosis are presented based on the study of 118 human cadavers (55 male, 63 female). The Martin-Gruber anastomosis was found in 25 (21.2%) of the 118 cadavers. It occurred in 11 (20%) of the 55 male cadavers (4 bilateral, 7 unilateral; 5 left and 2 right) and in 14 (22.2%) of the 63 female cadavers (2 bilateral, 12 unilateral; 8 left and 4 right). Therefore, the Martin-Gruber anastomosis was found in 31 (13.1%) of the 236 upper limbs. According to a recent classification (Rodriguez-Niedenfuhr et al., 2000), pattern I was found in 29 cases (93.5%), corresponding to Type A in 13 (41.9%), Type B in 3 (9.7%) and Type C in 13 (41.9%), whereas pattern II was found in 2 cases (6.5%), both being a duplication of Type IC. Intramuscular Martin-Gruber anastomosis was a single anastomosis that originated in all cases from the anterior interosseous nerve (pattern IC) and then passed through a muscle bundle of the flexor digitorum profundus and behind the ulnar artery to join the ulnar nerve as a single connecting branch. It did not send branches to the flexor digitorum profundus. This intramuscular course was observed in 3 of the 13 cases of Type C anastomosis (23.1%) or 3 cases out of 31 Martin-Gruber anastomoses (10%). PMID- 11877793 TI - Congenital undescended scapula (Sprengel deformity): a case study. AB - This study describes a congenital deformity of the shoulder joint in a 5-year-old female, associated with an undescended scapula and the presence of an omovertebral bone. Dissections on 16 cadavers were done to identify structures in danger of injure before operating on the patient. Surgeons need to take specific note of the dorsal scapular nerve. PMID- 11877794 TI - Variation in the origin of the inferior alveolar nerve. AB - The inferior alveolar nerve and maxillary artery were studied in 40 human heads through infratemporal dissection; in one specimen, a unusual variation in the origin of the inferior alveolar nerve and its relationship with the surrounding structures was observed. The inferior alveolar nerve originated from the mandibular nerve by two roots and the second part of the maxillary artery was incorporated between them. An embryologic origin of this variation and its clinical implications is discussed. Because the maxillary artery ran between the two roots of the inferior alveolar nerve, and the nerve was fixed between the foramen ovale and mandibular foramen, tension and compression of the nerve from arterial pulsation could cause mandibular neuralgia. PMID- 11877795 TI - Median (third) occipital condyle. AB - Abnormalities of the craniovertebral junction, most of which are associated with the occipital bone, are of interest not only to anatomists but also to clinicians because many of these deformities produce clinical symptoms. The occipital bone in humans is formed by the fusion of the sclerotomes corresponding to the roots of the hypoglossal nerve. In lower vertebrates, the cranial half of the first cervical sclerotome forms a separate bone between the atlas and occipital bone, the proatlas; in man it may contribute to anomalous structures around the foramen magnum. One such structure, a median occipital condyle, which exhibited an articular facet for the odontoid process, was observed in an adult male skull at the anterior margin of the foramen magnum. This anomaly is discussed in light of available literature. PMID- 11877796 TI - Traditions and peculiarities of clinical anatomy education in Russia. AB - The Russian experience in clinical anatomy education is described in this article. Such training is provided by the Department of Operative Surgery and Topographical Anatomy both during the pregraduate (undergraduate) period for medical students and in the postgraduate period for interns, residents, physicians, and surgeons of different specialties. The teaching of clinical anatomy in the pregraduate period occurs in combination with the study of operative surgery and follows the study of gross anatomy in the Department of Human Anatomy and microscopic anatomy in the Department of Histology, Cytology and Embryology. PMID- 11877797 TI - Teaching gross anatomy using living tissue. AB - Embalmed cadaver tissues and organs that are dissected in gross anatomy laboratories lack many characteristics of fresh or living tissues. The purpose of this study was to assess the educational value of allowing first-year medical students to experience first-hand the color, texture, delicacy and other qualities of living porcine tissues and organs that are similar to those of human tissues. Guided by a laboratory protocol, medical students palpated and inspected organs of the opened thorax, abdomen and pelvis of anesthetized pigs on pulmonary ventilators. The learning experience was rated highly by the students as well as by the participating faculty. A further review of the medical students' later experiences at autopsies and in surgical clerkships showed that the living-tissue experience in their gross anatomy course represented a large part of their medical school exposure to unembalmed tissues. PMID- 11877798 TI - The relationship between premedical coursework in gross anatomy and histology and medical school performance in gross anatomy and histology. AB - Many premedical students enroll in courses whose content will be encountered again during their medical education. Presumably, students believe this practice will lead to improved academic performance in corresponding medical school courses. Therefore, this study was undertaken to determine whether a premedical gross anatomy and/or histology course resulted in increased performance in corresponding medical school courses. A second aim of the study was to examine whether the type of premedical gross anatomy and/or histology course differentially affected medical school performance. A survey that assessed premedical gross anatomy and histology coursework was administered to 440 first year medical students. The results from this survey showed that students with premedical gross anatomy (n = 236) and/or histology (n = 109) earned significantly more points in the corresponding medical school course than students without the premedical coursework (P < 0.05). Analysis of premedical course types revealed that students who took a gross anatomy course with prosected specimens (n = 35) earned significantly more points that those students without premedical gross anatomy coursework (P < 0.05). The results from this study suggest: 1) premedical gross anatomy and/or histology coursework improves academic performance in corresponding medical school courses, and 2) a premedical gross anatomy course with prosected specimens, a specific type of undergraduate course, significantly improves academic performance in medical gross anatomy. PMID- 11877799 TI - Approach to the educational opportunities provided by variant anatomy, illustrated by discussion of a duplicated inferior vena cava. AB - Variant anatomy recognized during routine cadaveric dissection in the first year of medical school offers great learning potential by allowing students to gain enhanced understanding of an array of important subjects. It provides a framework for reviewing common morphology and embryogenesis of the structure in question, and through the help of appropriate faculty, yields insight into the potential medical, radiologic, and surgical implications. The frequency of clinically important anatomic variation is high enough to allow the gross anatomy laboratory to serve as an excellent teaching platform in this regard. Through anatomy, the student is introduced to the concept of patient individuality, and to the individualization of medical and surgical therapies. Recently, one of the variations encountered in our lab was a duplicated inferior vena cava. We describe our approach to such findings through a systematic discussion of the anatomy and embryology, as well as the radiologic and clinical correlates. PMID- 11877800 TI - Pharmacological properties of nicotinic acetylcholine receptors in isolated Locusta migratoria neurones. AB - Mechanically dissociated neuronal cell bodies from the thoracic ganglia of Locusta migratoria were viable in culture conditions for up to 2 days and were voltage-clamped to record the effects of GABAergic drugs and physostigmine on the membrane conductance and ACh responses of the dissociated cells. Bicuculline, hydrastine, and gabazine inhibited the EC50 ACh responses of the cells. Both bicuculline and hydrastine were full inhibitors of the ACh responses but gabazine behaved as a partial inhibitor. Bicuculline, hydrastine, and gabazine inhibited the ACh responses in a non-competitive and voltage-independent fashion, suggesting that they are allosteric inhibitors of locust nicotinic ACh receptors. Physostigmine activated currents when applied onto isolated locust neurones. The responses activated by physostigmine were inhibited competitively by tubocurarine, which indicates that physostigmine interacts with the ACh site of locust nicotinic ACh receptors. However, maximal concentrations of physostigmine elicited currents of smaller amplitudes to those evoked by maximal ACh concentrations. Single-channel recordings suggest that the partial efficacy of physostigmine may reflect the low frequency of opening of physostigmine-induced single currents relative to that of ACh-single currents. PMID- 11877801 TI - Motion detectors in the locust visual system: From biology to robot sensors. AB - Motion detectors in the locust optic lobe and brain fall into two categories: neurones that respond selectively to approaching vs. receding objects and neurones that respond selectively to a particular pattern of image motion over a substantial part of the eye, generated by the locust's own movements through its environment. Neurones from the two categories can be differentiated on the basis of their response to motion at a constant velocity at a fixed distance from the locust: neurones of the first category respond equally well to motion in any direction whereas neurones in the second category respond selectively to one preferred direction of motion. Several of the motion detectors of the first category, responding to approaching objects, share the same input organisation, suggesting that it is important in generating a tuning for approaching objects. Anatomical, physiological, and modelling studies have revealed how the selectivity of the response is generated. The selectivity arises as a result of a critical race between excitation, generated when image edges move out over the eye and delayed inhibition, generated by the same edge movements. For excitation to build up, the velocity and extent of edge motion over the eye must increase rapidly. The ultrastructure of the afferent inputs onto the dendrites of collision sensitive neurones reveals a possible substrate for the interaction between excitation and inhibition. This interpretation is supported by both physiological and immunocytochemical evidence. The input organisation of these neurones has been incorporated into the control structure of a small mobile robot, which successfully avoids collisions with looming objects. The ecological role of motion detectors of the second category that respond to image motion over a substantial part of the visual field, is discussed as is the input organisation that generates this selective response. The broad tuning of these neurones, particularly at low velocities (<0.02 degree/s), suggests they may have a role in navigation during migratory flights at altitude. By contrast, their optimum tuning to high-image velocities suggests these motion detectors are adapted for use in a fast flying insect, which does not spend significant time hovering. PMID- 11877802 TI - Signal processing in a simple visual system: the locust ocellar system and its synapses. AB - The neurons with the widest axons that carry information into a locust brain belong to L-neurons, the large, second-order neurons of the ocelli. L-neurons play roles in flight control and boosting visual sensitivity. Their morphology is simple, and their axons convey graded potentials from the ocellus with little decrement to the brain, which makes them good subjects in which to study transmission of graded potentials. L-neurons are very sensitive to changes in light, due to an abnormally high gain in the sign inverting synapses they receive from photoreceptors. Adaptation ensures that L-neurons signal contrast in a light signal when average light intensity changes, and that their responses depend on the speed of change in light. Neurons L1-3 make excitatory output synapses with third-order neurons and with L4-5. These synapses transmit tonically, but are unable to convey hyperpolarising signals about large increases in light. Graded rebound spikes enhance depolarising responses. L1-3 also make reciprocal inhibitory synapses with each other and transmission at these decrements so rapidly that it normally requires a presynaptic spike. The resolution with which graded potentials can be transferred has been studied at the inhibitory synapses, and is limited by intrinsic variability in the mechanism that determines neurotransmitter release. Electron microscopy has shown that each excitatory connection made from an L-neuron to a postsynaptic partner consists of thousands of discrete synaptic contacts, in which individual dense-staining bars in the presynaptic neuron are associated with clouds of vesicles. Acetylcholine is likely to be a neurotransmitter released by L-neurons. PMID- 11877804 TI - Impact of descending brain neurons on the control of stridulation, walking, and flight in orthoptera. AB - Orthopteran insects (crickets, bushcrickets, and acridid grasshoppers) are preferred preparations for the study of the central nervous mechanisms that underlie behavior. Many of their behaviors are based on central rhythm-generating circuits located in the ganglia of the ventral nerve cord. Activities of these circuits must be coordinated and adapted to the behavioral context by sensory information, which can derive from proprioceptive or exteroceptive inputs. Information from various sensory modalities converges in yet unidentified "higher brain centers" that integrate and transform it into activity patterns across populations of descending brain neurons (DBNs). Transmission of "decisions" to the thoracic motor centers leads to adjustment of their functions in order to fit the sensory situation encountered. A number of unique DBNs has been identified by morphological and physiological criteria and their role in controlling aspects of specific behaviors has been the subject of various studies. Their functions range from "switch-like" transmitters of brain output to complex integration units for sensory inputs of various modalities and their appropriate insertion into the ongoing activities of the thoracic rhythm generators. This paper highlights some of the characteristics of DBNs by focussing on three motor behaviors: stridulation, a stereotyped behavior that seems to be mainly controlled by command-like DBNs; walking, a plastic behavior whose various parameters must continuously be adjusted to a changing sensory environment; and flight, in which the information for course corrections encoded for by different types of DBNs is transformed to match the rhythmic activity of the flight oscillators before it affects the respective motoneurons. PMID- 11877803 TI - Developmental changes in the structure and function of the central olfactory system in gregarious and solitary desert locusts. AB - Desert locusts are guided by olfactory cues in different behavioural contexts. In order to understand the basis for the variable olfactory guided behaviour displayed by different developmental stages and by solitary and gregarious locusts, we investigated their central olfactory system with neuroanatomical and neurophysiological methods. The primary olfactory centre of the brain, the antennal lobe (AL), increases in size during development due to an increased number and size of glomeruli. These glomeruli are innervated by a constant number of projection neurons that display increased dendritic arborizations during the development of the locust. The anatomical parameters do not differ between gregarious and solitary locusts. In parallel with the observed neuroanatomical changes, neurophysiological changes in response spectra and response specificity of AL neurons were found. During development, the percentage of neurons responding specifically to aggregation pheromone components decreases, whereas an increase in both pheromone-generalists and plant-pheromone generalist neurons is observed. The percentage of neurons responding to green leaf volatiles, however, remains constant. A decrease in the number of nymph blend-specific neurons was also observed. Our data show that anatomical and physiological properties of the AL and its neurons to a large extent reflect the changes in olfactory guided behaviour during development and between phases. The majority of our results are also in accordance with findings that the number of olfactory receptor neurons increases during development, resulting in increasing convergence on AL neurons. PMID- 11877805 TI - Very low-magnification transmission electron microscopy. PMID- 11877806 TI - Cortical imaging through the intact mouse skull using two-photon excitation laser scanning microscopy. PMID- 11877807 TI - Inexpensive holder for hand-trimming resin-embedded tissues. AB - A simple and inexpensive mount for securely holding resin-embedded tissues for trimming prior to sectioning for light or electron microscopy is described. The unit is composed of a base plate from which a 12.7-mm drill chuck protrudes. A tissue block is clamped into a microtome chuck and that is mounted in the drill chuck. A standard dissecting microscope is placed over the unit and used to magnify the block for trimming. Total cost of materials is estimated to be $20-40 USD. PMID- 11877808 TI - Novel method for the plan-view TEM preparation of thin samples on brittle substrates by mechanical and ion beam thinning. AB - A new preparation method has been developed in order to avoid the breaking of brittle samples for plan-view TEM investigation during and after mechanical and ion beam thinning. The thinning procedure is carried out on a reduced size piece of the sample (about 1.6 x 0.8 mm(2) or about 1-1.6 mm diameter) that is embedded into a 3-mm-diameter Ti disk, which fits the sample holder of the TEM. The small sample size and the supporting metal disk assure the mechanical stability and minimize the possibility of breaking during and after the preparation: The Ti disk is placed on adhesive kapton tape, a cut piece of the sample is put into the slot of the disk, pressed onto the tape and embedded with glue. The tape keeps the parts in place and in the same plane, keeps the sample surface safe from the embedding glue and can be removed easily after the glue solidifies. Subsequently, the embedded sample is thinned from the rear by well-known mechanical and ion beam techniques until electron transparency. This simple solution lowers the risk of failed sample preparation remarkably and makes it possible to reduce the thickness of the sample to about 50 microm by mechanical thinning. As a result, dimpling becomes unnecessary and low angle ion milling gives a large transparent area for TEM. Its efficiency has been proved by successful preparation of numerous thin film samples on Si, sapphire, and glass substrates. The method is compatible with the widespread cross-sectional thinning procedures, and can be easily adopted by TEM laboratories. PMID- 11877810 TI - Cytochemical techniques for zinc and heavy metals localization in nerve cells. AB - Zinc is one of the most abundant oligoelements in the living cell. It appears tightly bound to metallothioneins, loosely bound to some metalloproteins and nucleic acids, or even as free ion. Small amounts of zinc ions (in the nanomolar range) regulate a plentitude of enzymatic proteins, receptors, and transcription factors; thus, cells need accurate homeostasis of zinc ions. Some neurons have developed mechanisms to accumulate zinc in specific membrane compartments ("vesicular zinc") which can be revealed using histochemical techniques. This article is a short report on the different direct-indirect experimental approaches for zinc and heavy metal detection in neurons. Substances giving a bright color or emitting fluorescence when in contact with divalent metal ions are currently used to detect them inside cells; their use leads to the so called "direct" methods. The fixation and precipitation of metal ions as insoluble salt precipitates, their maintenance along the histological process, and their demonstration after autometallographic development are essential steps for other methods, the so-called "indirect methods" (Timm and Danscher Neo-Timm methods). PMID- 11877811 TI - Recent advances in the histochemical staining of heavy metals by means of chelating agents, with special reference to cadmium. AB - This review discusses the way of improving the sensitivity and specificity of chelating agents used for the histochemical demonstration of trace amounts of heavy metals in cells and tissues. In the search for a more sensitive and specific agent, various chelating agents have been prepared and their staining properties for various metals examined. Of those developed 1-(2 benzothiazolylazo)-2-naphthol (BTAN), 2-(8-quinolylazo)-4,5-(diphenyl)imidazole (QAI), and 2-(5-bromo-2-pridylazo)-5-(diethylamino)phenol (BrPADAP) were far superior to conventional staining agents in staining sensitivity. This was particularly so for staining cadmium. When modifying chelating agents to increase staining sensitivity, hydrophobicity of the agent molecules, selection of substituent groups, and the position of the groups added to the agent molecules must be taken into consideration. The most frequently mentioned factors in a staining mechanism are complex-tissue attractive forces, and recent studies have made it clear that weak interactions, such as hydrogen bonds and hydrophobic interactions, contribute significantly. The Hansch pi values were a useful indication of the hydrophobicity of metal staining agents. The published values for stability constants of metal complexes were found to be unreliable guides for choosing a masking agent to block interfering metals. The polyphosphates, aminopolycarboxylic acids, and alkylamines were among the most useful masking agents for metal staining from a practical standpoint. PMID- 11877812 TI - Occurrence, distribution, and localisation of metals in cnidarians. AB - The Cnidaria are simple organisms that have remarkable physiological features susceptible to microscopic investigation. As a group they produce cnidae, the most complex intracellular organelles known, form symbioses with a range of unicellular algae, contain mucocytes that account for a very substantial fraction of their body mass, and form complex skeletal structures of calcium carbonate. This review summarises contributions dealing with the distribution and localisation of metals of physiological and pathological importance within soft tissues and skeletons. Whilst there have been detailed studies of microscale metal distribution, using X-ray microanalysis, in the stinging organelles or cnidocysts and in mucocytes, other cells such as symbiotic algae and the epithelial cells have received little attention. In the skeleton-producing scleractinian corals X-ray microanalysis has provided tenuous, but persistent, evidence of Ca associated with intracellular vesicles or granules in the skeletogenic epithelium, even though the investigations were technically limited. These observations may be germane to the intriguing and intransigent problem of the mechanism of coral calcification. Metal localisation in coral skeleton at the resolution of annual growth rings has been concerned with the validity of Sr/Ca and Mg/Ca ratios as thermometers for paleoclimatic studies. It is not clear whether these ratios are influenced primarily by environmental or biological parameters. Microscale analyses by X-ray microanalysis and ion microprobe indicate a much greater variability of metal ratios which suggests biological control of metal deposition. New data are provided on the elemental composition, measured by X-ray microanalysis, of cells and cell compartments in the coral Galaxea fascicularis and zooxanthellae in the anemone Aiptasia sp. New information is also presented on changing Ca/Sr ratios at the skeletal interface in Galaxea fascicularis. PMID- 11877814 TI - Localization of metals in cells of saprophagous soil arthropods (Isopoda, Diplopoda, Collembola). AB - This review summarizes results on the intracellular distribution of metals in cells of woodlice (Isopoda), millipedes (Diplopoda), and springtails (Collembola), which are three major groups of saprophagous arthropods contributing to the turnover of soil organic matter. Although the impact of metals and also metal pollution has inevitably been shown at levels of higher biological organization than subcellular mechanisms in these animal groups, the aim of this review is to focus exclusively on storage sites and aspects of intracellular metal metabolism. Thus, methodologically, results obtained by microscopical techniques such as histochemistry, X-ray microanalysis, energy filter transmission electron microscopy, or laser microprobe mass spectrometry were given preference. Results from atomic absorption spectrophotometry of cellular fractions were kept to a minimum. In all three taxa, the main intracellular metal storage sites are various types of "granules" which are widely distributed throughout cell types associated with the digestive system. PMID- 11877813 TI - Cellular and subcellular distribution of metals in molluscs. AB - The cellular processes involved in metal metabolism in molluscs are reviewed, with emphasis on the contribution of microscopy (AMG, ARG, EPMA, and SIMS) to both basic research of metal cell biology and applied environmental research. In molluscs, metal uptake may occur by facilitated diffusion, active transport, or endocytosis, and can be enhanced by MT synthesis or formation of mineralized granules. In aquatic molluscs, gills constitute a key interface for dissolved metal uptake, where metals are bound to MT, incorporated into lysosomes, and released basally towards the blood plasma and circulating hemocytes. However, particulate metal uptake is mainly achieved via the digestive tract by endocytosis; further metals are transferred first to lysosomes and then to residual bodies, especially in the digestive cells of the digestive gland. Additionally, metals can be accumulated selectively in specific cell types. As ligands pools differ from cell to cell, different metals may be retained in different cell types. Class "a" metals are localized in cells with granules composed of carbonate, oxalate, phosphate, and sulfate (oxygen donors), whereas "b" metals are associated with those cell types rich in sulfur and nitrogen ligands (sulfur donors). In molluscs, oxygen donors occur in connective tissue calcium cells and basophilic cells, whereas sulfur donors are present in digestive cells, podocytes, nephrocytes, and rhogocytes. Hemocytes, which constitute the most relevant system for metal transport between tissues, move around the body and may penetrate tissues and remove metals from the inner medium to be accumulated in lysosomes as nondigested products. Rhogocytes also participate in metal mobilization, accumulation, and release. The assessment of metal levels in target cells of sentinel molluscs by microscopic techniques provides an early-warning measure, with promising applications as an exposure biomarker for environmental monitoring programs. PMID- 11877815 TI - [Interventional magnetic resonance imaging]. PMID- 11877816 TI - [Use of intraoperative magnetic resonance imaging in neurosurgery]. PMID- 11877817 TI - [Gene amplification methods in viral diagnostics]. PMID- 11877818 TI - [The cause of occupational urticaria in hands]. PMID- 11877819 TI - [Giving birth to a dead fetus]. PMID- 11877820 TI - [On labor analgesia]. PMID- 11877821 TI - [Fast speed?]. PMID- 11877822 TI - [Colonoscopies]. PMID- 11877823 TI - [Patch testing should be controlled by authorities]. PMID- 11877824 TI - [Visual impairment is increasing]. PMID- 11877825 TI - [Sarcomere gene mutations--a cause of hypertrophic cardiomyopathy]. PMID- 11877826 TI - [Do vaccines cause allergy?]. PMID- 11877827 TI - [Statins for prevention of stroke?]. PMID- 11877828 TI - [Classification of juvenile rheumatoid arthritis becoming more precise]. PMID- 11877830 TI - [First Finnish phytosterolemia families have been identified]. PMID- 11877831 TI - [Serious complications following sinusitis in children]. PMID- 11877832 TI - [The upheaval of endoscopic surgery]. PMID- 11877829 TI - [Acquired color vision defects]. PMID- 11877833 TI - [Arthroscopic surgery of lower extremity joints]. PMID- 11877834 TI - [Gynecological endoscopic surgery]. PMID- 11877835 TI - [Laparoscopy--an increasingly popular method for abdominal surgical procedures]. PMID- 11877836 TI - [Endoscopy in urology]. PMID- 11877837 TI - [Light for the people]. PMID- 11877838 TI - [Selective surgery for varicose veins--let's preserve the healthy vessels]. PMID- 11877839 TI - [Diagnostics and therapy of amyloidosis]. PMID- 11877841 TI - [Amifostine as a supporting treatment during cancer therapy]. PMID- 11877840 TI - [Treatment of renovascular hypertension with balloon dilatation and stenting]. PMID- 11877842 TI - [Tropical fishes suitable pets for an allergic person?]. PMID- 11877843 TI - [Serositis]. PMID- 11877844 TI - [Hormonal treatment in women and thrombosis]. PMID- 11877845 TI - [Endoscopic retrograde cholangiopancreatography]. PMID- 11877846 TI - [Prevention of the first-episode psychosis: a rational basis for the second large progress step in psychiatry]. PMID- 11877847 TI - [A long-acting adrenergic beta-sympathomimetics has its place in the treatment of many asthmatic patients]. PMID- 11877848 TI - [Risk factors, prevention and prognosis of febrile seizures]. PMID- 11877849 TI - [Epoxy resin allergy caused by immersion fluid]. PMID- 11877850 TI - [What should be the target of therapy in the adult-onset diabetes mellitus?]. PMID- 11877851 TI - [Drug therapy of hyperglycemia and the prognosis of adult-onset diabetes mellitus]. PMID- 11877852 TI - [Assessing the individual risk of atherosclerosis]. PMID- 11877853 TI - [Treatment of dyslipidemias and its benefits]. PMID- 11877854 TI - [Treatment of hypertension in patients with adult-onset diabetes]. PMID- 11877855 TI - [Treatment of hyperglycemia in adult-onset diabetes mellitus]. PMID- 11877856 TI - [Treatment of coronary artery disease in a patient with adult-onset diabetes mellitus]. PMID- 11877857 TI - [Do all the patients with adult-onset diabetes mellitus need acetylsalicylic acid?]. PMID- 11877858 TI - [Bactericidal activity of biapenem against various bacteria]. AB - The bactericidal activity of biapenem (BIPM), a new carbapenem agent, against Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa and Serratia marcescens was compared with those of imipenem (IPM), panipenem (PAPM), meropenem (MEPM) and ceftazidime (CAZ). The bactericidal activity of BIPM against S. aureus was equal to those of IPM, PAPM and MEPM. Against E. coli and K. pneumoniae, BIPM showed higher bactericidal activity than IPM and PAPM. Against P. aeruginosa, BIPM showed excellent bactericidal activity campared with IPM. The killing speeds of BIPM and IPM were obviously the most rapid among four carbapenems. BIPM showed a strong bactericidal activity against 5 species of bacteria including P. aeruginosa. PMID- 11877859 TI - [The problem of post-marketing surveillance planning on drugs for infectious disease]. AB - In principle, a new drug is approved via the assessments of safety and efficacy by the Ministry of Health, Labor and Welfare (MHLW). After approval, conduct of post-marketing surveillance is requested by law in order to assess the safety and efficacy of the drug in a large number of patients. Before initiating a surveillance, submission to MHLW of a document on the plan of survey is necessary in the purpose of inspection if it includes any ethical or regulatory problem or not. Through the inspections of the plans submitted during the period of April 1998 and March 2000, many problems have been pointed out. In this report, the author introduces some of the concrete problems noted in the plans on drugs for infectious diseases and shows some of the guidance made by the authority how to improve them. It is expected that such practical analyses of cases may be useful for future planning of post-marketing surveillance on drugs for infectious diseases. PMID- 11877861 TI - Trip to HIPAA compliance. AB - The trip to HIPAA compliance can be unwelcome and burdensome. But, it must be made, however reluctantly. This article attempts to give some meaning to that trip and practical advice on how to arrive safely at your destination. The first part of the article will look at the new legislation providing an extension to compliance with the transactions and code set rules. The second part will address specific compliance strategies. PMID- 11877862 TI - Founding college of osteopathic medicine continues tradition. PMID- 11877860 TI - "Opportunity at 'sunset': Missouri relaxes its certificate of need requirements for ambulatory surgery centers". PMID- 11877864 TI - On field evaluation and management of head and neck injured athletes. AB - Cervical spine injuries, although relatively uncommon, can be devastating. Proper and prompt management is absolutely essential, even though the complexity of the brain and the lack of objective signs and symptoms make injury assessment difficult. Being prepared for head and neck injury is key to its on-field management. In the event of an injury, the equipment needed to evaluate, and if necessary, to transport the injured athlete must be available. The most important objective of on-field management of head and neck injuries is to prevent further damage and recurrent injury. PMID- 11877863 TI - Chest pain evaluation: a common clinical problem. AB - The clinical approach described here initially utilizing a careful focused history and physical examination, 12 lead ECG, as well as serum biomarkers, allows for an informed initial impression of the chest pain patient. Based upon the findings the practitioner can then initiate appropriate therapy or further testing based on this solid database. The therapy can vary from simple medical therapy involving aspirin and beta blockers to an invasive evaluation potentially involving percutaneous coronary interventions. If further non-invasive evaluation is chosen, then the choices for testing are as outlined here. The guidelines listed will help in selecting the appropriate test for the individual patient. We believe the approach used in this paper will help insure high quality cost effective care of the chest pain patient. PMID- 11877865 TI - Management of the athlete with low back pain. AB - The evaluation of an athlete with LBP using the classification system proposed by Delitto et al has been outlined. For outpatient orthopaedic practice, evidence in the literature is available documenting the reliability and the effectiveness of treatment guided by TBC. This classification system provides framework for the clinician to evaluate athletes with LBP because it investigates the presence of serious pathology, considers the severity of the disease process, and provides matched treatment based on the athlete's clinical presentation. When treating athletes with episodes of acute LBP, pain modulation and return to daily function are the primary treatment goals. When treating athletes with episodes of chronic LBP, return to sport and prevention of recurrence are the primary treatment goals. PMID- 11877866 TI - Back pain in the young athlete. AB - Back pain in the pediatric athlete is rare compared to the adult population. However, the diagnosis should be accurate as most have specific etiologies and treatment. Nearly 50% of the time, the cause of the pain is an injury to the pars interarticularis. A thorough history and careful physical examination should guide the physician to a working differential diagnosis. Based on this, appropriate laboratory and imaging work-up is pursued to arrive at a timely diagnosis and subsequent treatment regimen. PMID- 11877868 TI - Lumbar disc injury in the athlete. AB - The lumbar spine is a highly vulnerable area for injury in a number of different sports. And while the incidence is significant and the time lost may be significant, probably the most important problems are related to the fear of spinal injuries and the need for a therapeutic plan. Lumbar pain is significant in many sports, but an organized diagnostic and therapeutic plan can prevent permanent injury, allowing full function and maximum performance. PMID- 11877867 TI - Spondylolisthesis in the athlete. AB - The diagnosis and appropriate management of spondylolysis and spondylolisthesis in the athlete can be challenging for the treating orthopaedic surgeon. With an increase in the number of skeletally immature athletes competing at more demanding levels of competition, an associated increase in the number of such cases may be seen. Management strategy is directed by the symptoms, the age of the patient, and the severity of the spondylolisthesis. PMID- 11877869 TI - Transient quadriparesis in the athlete. AB - A considerable amount of controversy persists regarding return-to-play criteria and the risk for more severe injury after an athlete experiences an episode of transient quadriparesis. Similarly, the implication of the presence of congenital stenosis in an athlete participating in contact sports elicits great debate in the literature in terms of the athlete's risk for neurologic injury. The relatively infrequent occurrence rate of both transient quadriparesis and permanent cervical cord injury make it difficult to predict with certainty whether or not an episode of transient quadriparesis is a risk factor for permanent neurologic injury. The decision-making process in determining player eligibility in the face of congenital stenosis or after a documented spinal injury is difficult and at times confusing. Every injury and athlete should be evaluated on an individual basis in terms of cause, symptoms, radiographic findings, and previous history. It is hoped that the guidelines for return-to play delineated in this article will help the physician and the athlete make an informed and rational decision regarding the criteria for and relative risks of returning to participation in a contact sport. PMID- 11877870 TI - Intradiscal electrothermal therapy for the treatment of chronic discogenic low back pain. AB - The early studies on IDET are very promising. IDET offers patients with chronic discogenic low back pain an option other than chronic pain [figure: see text] management or spinal fusion. Studies currently under way will help answer questions relative to mechanism of action, placebo effect, and biomechanical changes after treatment. The cost, morbidity, and currently observed degree of effectiveness make IDET an attractive alternative to spinal fusion for discogenic pain. This may be especially true for the young patient with preserved disc height, and patients with inoperable multilevel disease. The technology was designed to be used for a specific diagnostic subset of disc disorders by specialists skilled in performing intradiscal techniques who possess the ability to accurately diagnose and effectively manage patients with complex spinal disorders. Abuse of this ground-breaking technology can be avoided if patient selection criteria are carefully observed and only skilled, technically proficient physicians perform the procedure. PMID- 11877872 TI - Cervical disc disease in the athlete. AB - The purpose of this article is to review the basic principles of cervical disc disease, and to outline strategies for the evaluation and treatment of this disorder. It is also intended to outline some guidelines regarding the difficult return-to-play decision for athletes with cervical disc disease. PMID- 11877871 TI - Cervical burners in the athlete. AB - Back pain in the pediatric athlete is rare compared to the adult population. However, the diagnosis should be accurate as most have specific etiologies and treatment. Nearly 50% of the time, the cause of the pain is an injury to the pars interarticularis. A thorough history and careful physician examination should guide the physician to a working differential diagnosis. Based on this, appropriate laboratory and imaging work-up is pursued to arrive at a timely diagnosis and subsequent treatment regimen. PMID- 11877873 TI - MR imaging in cervical spine trauma. AB - Continual improvements in MR imaging, technology and MR imaging-compatible monitoring and fixation devices have allowed the incorporation of this relatively new imaging modality into standard algorithms for cervical spine trauma assessment. The ability of MR imaging to define the type of spinal cord injury, the cause and severity of spinal cord compression, and the stability of the spinal column is unmatched. The heavy reliance of the spinal surgeon on MR imaging for decisions regarding the type of therapy, the timing, the approach of surgical intervention, and for predicting patient outcome attests to the usefulness of this modality. PMID- 11877874 TI - Spinal deformity in the adolescent athlete. AB - Today more and more children with diagnoses of adolescent spinal deformity are participating in athletic competitions. Certain sports appear to have an increased association with spinal deformity, especially in the adolescent female population. Modern treatment techniques including bracing will allow participation in most athletic endeavors. Those treated with surgery need to have their postoperative care individualized. However, modern surgical techniques still allow continued participation in most activities. PMID- 11877875 TI - Epidemiology of back pain in the athlete. AB - Back pain and injuries are common complaints of athletes. The nature of the sport involved with the specific stresses that it places on the athlete's spine may play a role in the injuries incurred. These injuries may limit an athlete's function and hinder performance; however, the majority of these insults are self limiting and respond well to conservative measures. Based on the epidemiologic data, it appears that possibly the single most important step in reducing the back pain experienced by athletes would involve the performance of a structured back-strengthening program. Such a plan might reduce the most common diagnostic entities encountered by the athlete with back pain. PMID- 11877877 TI - What cognitive science tells us about the design of reports for consumers. AB - The health services literature many articles related to expanding and refining quality measures. But the rich body of empirical research on how people process information has rarely been applied to the challenge of presenting complex information about health care in ways that facilitate its comprehension and use. In this article, the authors review key findings from this research. Based on their review, the authors develop some general principles for presenting information and demonstrate their utility by assessing three Web sites that report performance data. PMID- 11877876 TI - The impact of a CAHPS report on employee knowledge, beliefs, and decisions. AB - As comparative health plan performance data become more available, large employers are increasingly interested in knowing about the value in providing this information to employees to help them choose a plan. The purpose of this study is to empirically examine some of the key assumptions about how disseminating Consumer Assessment of Health Plans Study (CAHPS) report cards may influence employee knowledge, attitudes, and choice. The study used a quasi experimental design, with preintervention and postintervention interviews with 958 employees. The findings indicate that exposure to the reports is related to having more information on how well the different plans perform on the CAHPS reporting categories. Furthermore, those who saw the report perceive the reporting categories to be more important in health plan choice than those who did not. Finally, the findings show that those who saw the report are more influenced by information sent by their employer than those who did not see the report. PMID- 11877879 TI - Charity care: do not-for-profits influence for-profits? AB - This study further examines whether not-for-profit hospitals exert pressure on for-profit hospitals to provide charity care and whether for-profit hospitals react differently than not-for-profit hospitals to managed care pressures and hospital competition in providing charity care. A two equation model is estimated using 1996 data from California hospitals. The results indicate that in mixed ownership markets, for-profit hospitals provide significantly less charity care as not-for-profit hospitals in the market provide more. Unexpectedly, study for profit hospitals were not more influenced by price competition than other hospitals with respect to charity care. Having a unique role in providing charity care may justify continuing tax exemption for not-for-profit hospitals and enhance interest in payment and other policies with regard to conversions to ensure that not-for-profit hospitals continue to be represented in market areas. PMID- 11877878 TI - Response to symptoms among a U.S. national probability sample of adults infected with human immunodeficiency virus. AB - Previous studies concerning disparities in Human Immunodeficiency Virus (HIV) services use among vulnerable groups did not control for specific clinical need for care such as symptom events. Using the Andersen Behavioral Model of Health Services Use, the authors determined whether minorities, women, and the less educated (vulnerable groups) were less likely to receive care for HIV symptoms. Persons enrolled in the HIV Cost and Services Utilization Study were asked whether they received care for their most bothersome symptom. Surprisingly, minorities and women were no more likely to go without care than other groups. Those with Medicaid, Medicare, private health maintenance organization (HMO) insurance, or no insurance were less likely to receive care for symptoms than those with private-non-HMO insurance. Vulnerable groups were no less likely to use services for HIV-related symptoms when need for care was considered. However, disparities may exist for symptom-specific care among HIV infected persons covered by public or HMO insurance. PMID- 11877880 TI - Declining employer-sponsored coverage: the role of public programs and implications for access to care. AB - Using data from the 1996/1997 Community Tracking Study household survey, this study examines the effects of public programs on the decision to take up employer coverage when offered versus enrolling in public coverage or being uninsured. The results show that among those with access to employer-sponsored coverage, low income persons living in states with more expansive eligibility for Medicaid were more likely to decline employer coverage in favor of public coverage, while low income persons in areas with public hospitals were more likely to decline coverage in favor of being uninsured. While persons who decline employer coverage in favor of public coverage maintain the same level of access to medical care, those who decline coverage in favor of being uninsured give up a considerable degree of access. Implications concerning policies to improve access to care for the uninsured are discussed. PMID- 11877881 TI - [Lipid disturbance of cell membranes in the etiopathogenesis of schizophrenia and affective disorders]. AB - The authors present information on the key role of lipid disturbances (mainly phospholipids) of neural membranes in the etiopathogenesis of schizophrenia and affective disorders. Current theories on membrane phospholipid disturbances in the etiopathogenesis of schizophrenia and affective disorders are connected to the idea that the primary metabolic disorder in both these disorders is due to the altered activity of phospholipase AZ, which in turn brings about disturbances in other lipid elements. Especially the functional insufficiency of some necessary fatty acids, as well as disturbed prostaglandin secretion has been shown in these disorders. In the recent years evidence has been collected on the action of pharmacologic substances used in these disorders on the lipid enzyme metabolism. PMID- 11877882 TI - [The psychopathological symptoms of the preclinical stages of Alzheimer's disease]. AB - The aim of the undertaken studies based on many years' observation of the group of elderly people was to describe psychopoathological pictures typical of preclinical phase of Alzheimer disease (AD) as well as the changes in social functioning. The obtained data were to serve to work out the criteria making initial recognition of preclinical phase of AD possible. Estimation of direct degree--in the perspective of next few years--of the danger of dementia of Alzheimer type should simplify making a decision about including therapeutic activities. 204 persons have completed the five-year-observation (70% of the qualified). During the observation AD has developed at 19 persons, five of the examined at the beginning of the observation were described as "no disturbances of cognitive functions"--I according to the GDS scale and 14 were qualified as "slight impairment of cognitive functions"--GDS II. The basis for further conclusions was the comparison of the results obtained in the groups of people at whom, during the five-year-observation, in the examination with the GDS scale, no changes regarding cognitive functions have been observed, with those examined in whom dementia has been recognised. Statistic analysis has been done mostly for the results obtained before the appearance of the dementia. The conducted studies made it possible to verify the stated hypotheses and draw the following conclusions: preclinical period of AD is connected with intensification of psychopathological disturbances, especially depression; in the period preceding the development of AD usually slight disturbances of cognitive functions appear which are most often revealed during the particular examination of cognitive functions; clear decrease in social activity is typical of the preclinical phase of AD; people in premorbid period showed various disturbances but no symptom has been observed that would be pathognomic for the preclinical phase of AD; it is possible, on the basis of the examination by means of proper clinical scales, to predict the danger of dementia but it is necessary to take into account various elements of psychic state and social functioning. PMID- 11877883 TI - [Risperidone in the ambulatory treatment of behavior disorders in demented patients of Alzheimer's type: a retrospective analysis]. AB - Behavioural disturbances are common in the course of dementia in Alzheimer's disease (AD) and their treatment is usually difficult. Different pharmacological and non-pharmacological options are employed basing mainly on clinical experience, still the number of well-designed, controlled studies in the field is very small. Novel, atypical neuroleptics, including risperidone might potentially be one of these options, taking into account their good safety profile and clinical efficacy in closely related syndromes. We present the results of a retrospective analysis of 57 outpatients with behavioural symptoms complicating AD treated with risperidone, either alone or in combination with one of the acetylcholinesterase inhibitor (AchEI; donepezil or rivastigmine). Seventy five percent of patients treated responded to risperidone with the usual effective dose of 0.5-1 mg/day. The influence of risperidone treatment on behavioural symptomatology was irrespective to the use of AchEI and equally well safe in both groups. The clinical response to the treatment was seen usually within first 2-3 weeks, those who did not respond early tended not to respond later on as well. Additionally, if not responding to low doses of risperidone (0.5-1 mg/day), patients usually did not respond to higher doses or could not tolerate them, mainly due to emerging extrapyramidal symptoms (EPS). Low doses of risperidone were well tolerated, with the fraction of patients experiencing EPS not achieving 10%. EPS observed, were dose dependent and tended to appear if the dose acceleration was fast. We then recommend low doses of risperidone and its slow titration if needed. PMID- 11877884 TI - [Emotional states of army recruits on the day of their recruitment to the army]. AB - Adaptive stress present in army recruits is the result of a permanent and still enhanced by the media stereotype which brings about the experience of deep emotional states. The study, based on empirical representative material of a group of 552 soldiers, presents the type of stress and emotions accompanying it experienced as well as presents the postulates of the soldiers examined on the possibility of stress reduction. In the conclusion, the author implies on the necessity of the introduction of a programme which would reduce adaptive stress amongst soldiers and would also improve the image of army service in the social viewpoint. PMID- 11877885 TI - [The evaluation of the quality of life among professional soldiers with diagnosed alcohol addiction syndrome]. AB - Presented paper is the first estimation probe in Poland of the "quality of life" of alcohol addicted people. The group of 70 professional soldiers that had been undergo treatment in Military Hospital the Section of Psychiatric Clinic in Cracow in years 1996-1998 has been investigated as alcohol addicted syndrome. For the research it has been used: the questionnaire with sociodemographis and epidemiological data, Polish version questionnaire SF-36 for "quality of life" research and Michigan Alcoholism Screening Test (MAST)--the questionnaire for estimation alcohol problems. The main aim of investigation was to recognise the influence of alcohol addicted syndrome that impact on "quality of life" estimation, range changes and their dynamics that depend on alcohol addicted syndrome degree, in age, military degree. The exploration aim was some sociodemographis and epidemiological data analysis. Results confirm the lower "quality of life" within estimated group. All ingredients Q1 have decreased, and the highest differences related such ingredients as: "physical limits in role perform", "social function", "emotion limits in role perform" so that "social activity sphere" creation ingredients. The estimation hypothesis related to QL decrease together with addiction degree (numbers of MAST) and decrease of "quality of life" together with age within estimated group have been not confirmed. The analysis of chosen sociodemographis variables has shown the significant variation, relative to QL and: "sexual live", "alone drinking" and "conflict". The differences between peoples who are alcohol addicted and non habitual drinking alcohol peoples has been confirmed within range; "frequency of alcohol drinking", "quantity of single drinking alcohol", "frequency to be drunk", "reasons of drinking", "behaviour after drinking", "suffering after drinking", "continuos drinking" or "alcohol tolerance". PMID- 11877886 TI - [Biological basis of psychiatry]. AB - Psychiatry had retained the status of art rather than science for longer than other branches of medicine. However, the views that psychiatric disorders have material basis, existing since antiquity, had eventually been accepted by modern medicine, in spite of troubles connected with the rise of analytical psychiatry. Presently, there are no doubts that structural and functional integration of the brain is a prerequisite of psychical health. The psychical processes are presently regarded as neurobiological phenomena, and the technical progress, particularly brain imaging techniques, allow to investigate, with objective methods, even phenomena such as consciousness, that for long remained in the domain of philosophy. The applied success of biological psychiatry is the introduction of effective psychotropic drugs. It cannot be forgotten, however, that psychotherapeutic methods (which also have neurobiological roots), are as indispensable as psychopharmacotherapy in a modern psychiatric clinic. PMID- 11877887 TI - [A feeling of coherence and delayed sequelae of post-traumatic stress in persons persecuted for political reasons in Poland in the years 1944-56]. AB - The paper presents the results of research of 100 persons persecuted for political reasons in Poland in the years 1944-56. The research aims to study the relations between the level of Sense of Coherence and delayed effects of post traumatic stress disorder in personal functioning (which is manifested by anxiety, depression, lack of meaning of life, and low well-being), and in interpersonal functioning. Introduction of this article presents the problems connected with delayed effects of post-traumatic stress disorder, and the general principle of Antonovsky's salutogenic concept also characterizes the sense of coherence category (SOC). SOC is a generalized, long-lasting feeling of confidence that the world is comprehensible (internal and external environments are structured, predictable, and explicable). People with a high level of SOC see their life as having some purpose and that their life is worthy of personal investments, and see themselves as having the personal resources to cope with the difficulties. The data was collected with using: The Sense of Coherence Questionnaire, PTSD-Interview, Beck Depression Inventory, State-Trait Anxiety Inventory, Purpose in Life Test, The Psychological General Well-Being (PGWB) Index, and The Scale of Interpersonal Relationships. The results indicate that the strength of the sense of coherence is negatively correlated with level of anxiety, depression, enmity syndrome, and positive with meaning of life, psychological well-being, and pro-social relationships. The research confirmed the important role of a sense of coherence (SOC) in modifying experience of delayed consequences of post-traumatic stress in victims of political persecution. PMID- 11877888 TI - [Symptoms of paranoid schizophrenia and anxiety: a dynamic analysis]. AB - The paper analyses the connection between anxiety and different symptoms of paranoid schizophrenia. In the study 66 patients admitted to hospital with an episode of paranoid schizophrenia were examined by a set of tests. Exacerbation of anxiety was measured by State-Trait Anxiety Inventory (STAI), whilst symptoms of schizophrenia--by the Brief Psychiatric Rating Scale (BPRS) and Scale for Assessment of Positive and negative Symptoms (SANS < SAPS). Statistically important correlations between anxiety and the majority of positive symptoms were observed. On the other hand, correlations between negative symptoms and anxiety experienced by the sick appeared variously. PMID- 11877889 TI - [Gender related prognostic factors in the first admission of DSM-III schizophrenic patients]. AB - In the group of the 80--DSM-III--first schizophrenic out patients we evaluated differences between men and women in predictive factors. First symptoms of illness were manifested later in women than in man. What's more women achieved better results in social functioning as well in global assessment, as in partial criteria. Work was the only domain in which there wasn't any differences in favour of women. Women had a lower rate of personality disorders. There were no differences in psychiatric symptoms severity. PMID- 11877890 TI - [Disorders of emotional control in schizophrenia and unilateral brain damage]. AB - Although, emotions play a crucial role in schizophrenia, the changes in emotional dimension still remain controversial. The aim of our work was: 1) to compare the disorders of emotional control between the examined groups: S--non-chronic schizophrenic patients (n = 50), CS--chronic schizophrenic patients (n = 50), N- healthy controls (n = 50), R--right brain-damaged patients (n = 30), and L--left brain-damaged patients (n = 30), 2) to assess a level of impairment of emotional control, its relation to lateralised hemisphere damage and chronicity of schizophrenic process. All psychiatric subjects were diagnosed as paranoid schizophrenics according to DSM-IV criteria and were scored on the PANSS scale after four weeks of neuroleptic treatment. Brain-damaged patients were included if they experienced single-episode cerebrovascular accidents causing right or left hemisphere damage (confirmed in CT scan reports). The neurological patients were examined at least 3 weeks after the onset of cerebrovascular episode. Emotional control was assessed using Brzezinski Questionnaire of Emotional Control aimed at the evaluation of: 1) control in perception and interpretation of emotive situation, 2) emotional arousal, 3) emotional-rational motivation, and 4) acting caused by emotions. Our results revealed significantly greater impairment of emotional control in schizophrenics (chronic schizophrenics, in particular) compared to healthy volunteers. Chronicity of the schizophrenic process seemed to intensify emotional control impairment. Interestingly, no significant qualitative and quantitative differences in emotional control mechanism between unilateral brain-damaged patients and the control group were found. PMID- 11877891 TI - [Perception of chimeric faces in schizophrenia and right hemispheric dysfunction]. AB - The aim of this study was the assessment of cerebral specialisation in perception of emotional chimeric drawings in 50 non-chronic schizophrenics (S), 50 chronic schizophrenics (CS), 30 right brain-damaged inpatients (P), and 50 normal controls. All were marked right handers. The assessment was performed after a four-week treatment. Structure and intensity of schizophrenia symptomatology were scored on the PANSS scale. Happy-sad chimeric face drawings (David 1989) were viewed twice in free vision. A perceiver bias towards left hemiface of chimeric drawings (LHF bias) and sad bias were scored in all subjects. Subjects rated their mood at the time of testing on a visual analogue scale. The schizophrenics and right brain-damaged inpatients showed significantly weaker LHF bias compared with healthy subjects, which may suggest right cerebral hemisphere dysfunction in perception of emotional chimeric drawings. Moreover, chronic schizophrenic patients revealed significantly weaker LHF bias and sad bias compared to non chronic subjects. There was no correlation of left perceptual bias with clinical ratings: PANSS scale, MMSE, number of hospitalisations, neuroleptic dose, and current mood, which suggests stable properties of the perceptual deficit. PMID- 11877892 TI - [The ability of modifying cognitive deficits in the area of social functioning in chronic schizophrenic patients]. AB - The paper concerns the effectiveness of rehabilitation of chronically ill schizophrenic patients who participate in social skills training. The assumption was that the participation in a four-month training programme (as authored by R. P. Liberman) will improve emotional problem solving, ability to keep in norms and conflict solving as well as the ability of cause-effect thinking. 100 schizophrenic patients and their families participated in the study. Half of those studied participated in the social skills training programme and the half who did not, were the control group. The subjects were examined twice in the four months. The following tools were used: "Means Ends Problem Solving" by Platt and Spivack, "PANSS" by Kay and Fiszbein, "Social-demographic questionnaire". Before the study period the groups did not vary much in the aspect of interpersonal social skills solving. After the training, the participants benefited significantly in the effectiveness of their problem solving and other criteria improved as well. It was concluded that the study of a group of 100 chronic schizophrenic patients who participated in a four month social skills training programme has a significant effect on social skills problem solving. The improvement of those skills consisted of: interpersonal and emotional problem solving, prediction of the consequences of one's actions, cause-effect thinking and alternative thinking. PMID- 11877893 TI - Developing prosthetics to treat cognitive disabilities resulting from acquired brain injuries. AB - Persistent cognitive disabilities represent the most troublesome consequences of acquired brain injury. Although these problems are widely recognized, few neuroprosthetic efforts have focused on developing therapeutic strategies aimed at improving general cognitive functions such as sustained attention, intention, working memory or awareness. If possible, effective modulation of these neuropsychologic components might improve recovery of interactive behaviors. The emerging field of neuromodulation holds promise that technologies developed to treat other neurological disorders may be adapted to address the cognitive problems of patients suffering from acquired brain injuries. We here discuss initial efforts at neuromodulation in patients in the persistent vegetative state and aspects of recent studies of the underlying neurobiology of PVS and other severe brain injuries. Innovative strategies for open-loop and closed-loop neuromodulation of impaired cognitive function are outlined. We discuss the possibilities of linking neuromodulation techniques to underlying neuronal mechanisms underpinning cognitive rehabilitation maneuvers. Ethical considerations surrounding the development of these strategies are reviewed. PMID- 11877894 TI - Significance of computed tomography mixed density in traumatic extra-axial hemorrhage. AB - Computed tomography (CT) mixed density of traumatic extra-axial hemorrhages (TEH) or the 'swirl sign' has been reported to correlate with active bleeding found at craniotomy and poor outcome. This study was done to test the hypothesis that mixed density of TEH detected by third or fourth generation CT correlated with the type of bleeding or clinical outcome. All cases of TEH operated at Detroit Receiving Hospital from 1991-1997 were reviewed for type of bleeding (active vs. not active; arterial vs. venous); Glasgow Coma Scale (severe 1-8, not severe 9 15); and Glasgow Outcome Score (1-3 poor; 4,5 good). CT density (CTD) of 51 cases with specific written documentation of bleeding type were then independently reviewed (SKS and MHR) and classified into TEH with mixed or high density. Data was analyzed using corrected Chi Square analysis, Fischer's Exact Test and Pearson's Correlation (SPSS 6.0). The Pearson Chi Square probability for correlation follows: CTD vs. active bleeding, 0.21; CTD vs. arterial, 0.41; CTD vs. severity, 0.57; and CTD vs. outcome, 0.81. No other statistical analysis identified a significant correlation, thus the null hypothesis could not be rejected. CT mixed density was not found to be correlated by more than chance with bleeding type, injury severity or outcome. Surgeon inaccuracy in documentation of bleeding type and use of later generation CT may account for the discrepancy between this and previous studies. Nevertheless, we conclude clinical exam and other published CT criteria are better indicators of injury severity and outcome. PMID- 11877895 TI - Vertebrobasilar dissection: a possible role of whiplash injury in its pathogenesis. AB - We reviewed 29 patients with vertebrobasilar dissections (VBD) to investigate the correlation between minor trauma and VBD and the clinical features of this trauma related condition. Mean age was 43 years, with a male predominance (male/female ratio was 25/4). Seventeen patients presented with subarachnoid hemorrhage (SAH), and 12 with ischemic symptoms. Two patients presenting with ischemia had extracranial VBD (V3 segment). Angiographically, aneurysmal dilatation was observed in most SAH patients (13 patients) in contrast to narrowing or occlusion in most ischemic patients (10 patients). Among the 12 SAH patients treated with coil embolization or conservatively, five died, whereas all ischemic patients recovered well with anticoagulation and/or antiplatelet therapy. Seven patients had received minor or trivial head/cervical trauma, due to whiplash injury, minor fall, or during exercise, which were identified to precede with the lapse of some time (a few minutes or days) the onset of symptoms. All of these patients presented with ischemic symptoms, and they were younger than the other ischemic or SAH patients. The site of vertebral artery dissection was intracranial in four cases, extracranial in one case, and combined in two cases at the level of the V3 segment and the origin of the posterior inferior cerebellar artery. However, no SAH occurred. These clinicopathological findings, i.e. ischemia and angiographic narrowing/occlusion, suggest that dissections were subintimal. Therefore, it is believed that this minor or trivial trauma may primarily cause subintimal dissection with luminal compromise, leading to ischemic symptoms, rather than subadventitial or transmural dissection with aneurysmal dilatation, leading to SAH. This lesion may also occur in younger patients with a favorable outcome. Careful note should be made of patient for the early recognition of this disorder. PMID- 11877896 TI - Limbus lumbar and sacral vertebral fractures. AB - We evaluated the fractures of the lumbar and sacral vertebral limbus by disc impingement at the peripheral ring apophysis in 23 adults associated with trauma in 16 of them. Lumbalgia, radicular pain and narrow canal symptoms are the presenting forms of this underdiagnosed pathology. CT is the best method of examination, while plain roentgenograms and MR are usually negative. Accurate diagnosis and surgical technique with larger exposure are needed to resect the fractured fragments and protruded disc material for decompressing the roots and the dural sac. Our results were very good on the majority of cases. PMID- 11877898 TI - Skull base trauma: diagnosis and management. AB - The singular anatomical relationship of the base of the skull is responsible for the particular problems that may arise after injury. Extensive dural laceration and severe neurovascular damage may accompany skull base injuries. Trauma to the anterior skull base is frequently related to the paranasal sinuses, and trauma to the middle and the posterior skull base usually affects the petrous bone. Injury to the anterior fossa including the paranasal sinuses may produce CSF leakage, damage the olfactory nerves, optic nerves, and orbita contents. Fractures may affect the carotid canal, injure the internal carotid artery and result in carotid-cavernous fistula. Trauma to the petrous bone may cause facial palsy and deafness, and CSF leakage with otorrhoea or paradoxal rhinoliquorrhoea. Trauma to the posterior fossa may lacerate the major venous sinuses, and affect the cranio cervical stability. Each one of these injuries will need a particular strategy. Decision making for management as a whole must consider all aspects, including the fact that these injuries frequently involve polytraumatized patients. Decisions regarding the timing of surgery and the sequence of the surgical procedures must be made with great care. Modern surgical techniques and recent technologies including functional preservation of the olfactory nerves in frontobasal trauma, visual evoked potentials, assisted optic nerve decompression, facial nerve reconstruction, interventional technique for intravascular repair of vascular injuries, and recent developments in cochlea implants and brain stem implants, all contributed significantly to improve outcome and enhance the quality of life of patients. This article reviews basic principles of management of skull base trauma stressing the role of these advanced techniques. PMID- 11877897 TI - Brain stem lesions after head injury. AB - There is little knowledge on the morphology of the brain stem in survivors of head injury, as CT fails to shown brain stem lesions, and neuropathological data is only available from autopsies. As magnetic resonance imaging (MRI) sheds new light on morphological lesions of the brain, the authors investigated 100 patients with a severe head injury. MRI was performed in a prospective study within the first seven days after head injury while the patients were still in coma and on ventilation. Relating the location of the lesions as depicted by MRI with the initial CT scan and outcome, death appeared to be closely linked to the phenomenon of bilateral pontine lesions. The extent of supratentorial lesions had no bearing on survival at all in the absence of brain stem lesions. Altogether the brain stem was affected in 52%. Obviously the occurrence of bilateral upper pontine lesions is of highest predictive value for a fatal outcome. Severe destruction of supratentorial white matter as demonstrated by MRI is not related to increased mortality, as long as the brain stem is spared. PMID- 11877899 TI - Vascular tunnel construction in the treatment of severe brain swelling caused by trauma and SAH. (evidence based on intra-operative blood flow measure). AB - Decompressive craniectomy with durotomy, is possible as a last resort therapy for severe traumatic brain swelling. Although the method successfully diminishes the ICP, partial or total vascular insufficiency occurs in the herniated part of the brain. The actual cause of the insufficiency is most likely due to the compression of the cortical veins and arteries supplying the herniated brain, caused by shearing and pressure forces between the dural edge and brain tissue. Furthermore venous congestion may induce edema in the protruding parts of the brain, thus further compromising neurone viability. The new surgical technique consists of a stellate type durotomy and the creation of a vascular tunnel around the main cortical veins and arteries, with the aim that the vessels do not become compressed by the dural or bone edge. The effect of the novel vascular tunnel technique was proven by measuring the blood flow of the protected and nonprotected veins with Doppler UH, intra-operatively. In the last two years 28 patients were operated on with this method. One case of edema was caused by SAH. All were in severe GCS 3 or GCS 4 status, with more than 30 mmHg ICP. In comparison with the traditional surgical and nonsurgical treatment, where the reported mortality rates are 80%-90% in these severe cases the mortality rate was reduced to 40%, and recovery (GOS 4, 5) rate also increased significantly. With this technique the ICP was significantly reduced and further edema and vascular insufficiency was prevented. This was due to protection of the arterial circulation and venous drainage of the herniated part of the brain, by the formation of a vascular tunnel at the durotomy edges. PMID- 11877900 TI - Relationship between brain temperature, brain chemistry and oxygen delivery after severe human head injury: the effect of mild hypothermia. AB - We studied brain temperature and the effect of mild hypothermia in 58 patients after severe head injury (SHI). Brain tissue oxygen tension (ptiO2), carbon dioxide tension (ptiCO2), tissuie pH (pHti) and temperature (T.br) were measured using a multiparameter probe. Microdialysis was performed to measure glucose, lactate, glutamate, and aspartate in the extracellular fluid. Mild hypothermia (34 degrees-36 degrees C) was employed in 33 selected patients who had persistent increased intracranial pressure (ICP > 20 mmHg). Mild induced hypothermia decreased brain oxygen significantly from 33 +/- 24 mmHg to 30 +/- 22 mmHg (p < 0.05). The ptiCO2 (46 +/- 8 mmHg) was also significantly lower during mild hypothermia (40.4 +/- 4.0 mmHg), p < 0.0001). The pHti increased from 7.13 +/- 0.15 to 7.24 +/- 0.10 (p < 0.0001) under hypothermic conditions. Induced hypothermia may protect patients from secondary ischemic events by lowering the critical ptiO2 threshold, reducing anaerobic metabolism, and decreasing the release of excitatory aminoacids. However, patients with spontaneous brain hypothermia on admission (Tbr < 36.0 degrees C) showed significantly higher levels of glutamate as well as lactate, compared to all other patients, and had a worse outcome. Spontaneous brain hypothermia carries a poor prognosis, and was characterized by markedly abnormal brain metabolic indices. PMID- 11877901 TI - Inhibition of human LDL oxidation by the neuroprotective drug l-deprenyl. AB - L-deprenyl (Selegiline) used in the treatment of Parkinson's and Alzheimer's disease also enhances longevity. Oxidized low density lipoprotein promotes atherosclerosis and is toxic to both vascular and neural tissue. The reported association between vascular dysfunction and neurodegenerative diseases prompted us to investigate the effect of l-deprenyl, a MAO-B inhibitor, on low density lipoprotein (LDL) oxidation. LDL was isolated from freshly collected blood and the kinetics of copper induced oxidation of LDL was monitored continuously by spectrophotometry. Oral administration (10 mg) or in vitro (2.8 to 84 microM) addition of l-deprenyl inhibited oxidation of LDL isolated from healthy men and post-menopausal women. This is the first report demonstrating that the antioxidant action of l-deprenyl may be antiatherogenic and cardioprotective. Such an action could contribute to reported extension of life span associated with long-term administration of the drug. In conjunction with inhibition of LDL oxidation, l-deprenyl is unique in that it demonstrates protective effects on both vascular and neuronal tissue. Prophylactic use of low doses of l-deprenyl may accord protection against vascular and neurodegenerative diseases associated with aging. PMID- 11877902 TI - Sympathetic mouth movements accompanying fine motor movements in chimpanzees (Pan troglodytes) with implications toward the evolution of language. AB - Some theories concerning the evolution of language include a gestural stage prior to glottogenesis. These theories propose that connections observed between fine motor movements of the hands and mouth may be responsible for the transfer of human language from one that was primarily gestural to one that is spoken. The fine motor manipulation of objects by five captive chimpanzees was examined to determine whether sympathetic mouth movements accompanied fine and gross motor movements. Sympathetic mouth movements were observed significantly more often during fine motor manipulation for all five participants. A significant increase in the presence of sympathetic mouth movements was observed in behaviors along a continuum of precision of manipulation from precision grip to gross motor manipulation without prehension. The results are discussed in regard to current theories of language evolution and neurological processes. PMID- 11877903 TI - Focal cerebral ischemia in rats produced by intracarotid embolization with viscous silicone. AB - Many factors contribute to the severity of neuronal cell death and the functional outcome in stroke. We describe an embolic model of focal cerebral ischemia in the rat that does not require craniotomy and is compatible with continuous measurement of regional CBF using multichannel laser Doppler flow (LDF) technique. Either a 22 microliters (large lesion) or 11 microliters (small lesion) bolus of viscous silicone was injected cephalad into the internal carotid artery. Upon injection, LDF decreased abruptly, most severely in the parietal cortex (-74% +/- 5%) in the large lesion and in the occipital cortex (-69% +/- 10%) in the small lesion model. Over the first hour, post-embolization LDF improved in most areas (e.g. -48% +/- 9% parietal, large lesion) but declined in the small lesion group in the occipital region (-81% +/- 8%). CBF measured by [C]14-IAP autoradiography 1 h post-embolization in the large lesion model demonstrated near-hemispheric ischemia (70% of hemisphere) with sparing of cingulate cortex. Autoradiography demonstrated that ischemia in the small lesion was largely cortical. Light microscopy of brains embolized with 11 microliters of dyed silicone showed filling of pial vessels with no silicone in the Circle of Willis or parenchyma. No animals in the large lesion group survived 24 h. Thirteen of 15 animals in the small lesion group survived for two weeks with resolution of initial hemiplegia, ocular asymmetry and weight loss. Hematoxylin eosin staining two weeks post-embolization showed signs of severe hypoxia and infarction. In conclusion, the intracarotid silicone embolization technique produces a titrable, reproducible permanent ischemic injury by blocking perfusion in the pial circulation, and is amenable to multisite monitoring with laser Doppler flowmetry. The smaller embolus produces cortical infarction with high rate of survival and neurological recovery. PMID- 11877904 TI - Experimental stereotactic gamma knife radiosurgery. Vascular contractility studies of the rat middle cerebral artery after chronic survival. AB - In vitro isometric small vessel myograph experiments and pathological investigations were performed on rat middle cerebral arteries. Thirty-four animals provided 68 normal vessels, six further rats had the endothelial layer mechanically removed from their 12 arteries. Eighteen animals received gamma knife irradiation to the middle cerebral arteries. Fifteen of these received 50 Gray, and three 25 Gray dose to the 50% isodose and the contralateral vessels offered 20 Gray and 15 Gray irradiated specimens. Survival times varied from 12 weeks to 18 months. In the acute stage, abolition of potassium-induced relaxation occurred as early as 24 h after irradiation whilst in one year this reaction seemed to recover and remained active to 18 months. The contraction response to prostaglandin F2 alpha was diminished at six weeks in the 50 Gray-irradiated vessels. However, from one year further reduction was seen and by 18 months this response was totally abolished. We demonstrated reduction of contractile capability of the irradiated normal vessels while the vessels remained patent. When using low irradiation dose there were no pathological changes even at 18 months, but marked physiological changes could be demonstrated. Different vessel wall functions appear to have different radiosensitivity, time course and capability for regeneration. PMID- 11877906 TI - Impairment of autoregulation following cortical venous occlusion in the rat. AB - Recent experiments showed an upward shift of the lower limit of autoregulation (AR) following photochemical occlusion of cortical veins in the rat. The goal of the present study was to prove the hypothesis that occlusion of cortical veins will be associated with impairment of the upper limit of autoregulation as well. In n = 28 Wistar rats unilateral frontoparietal cranial windows were drilled for transdural assessment of regional cerebral blood flow (rCBF) by laser Doppler scanning. The animals were allotted to two groups: (1) Group A (n = 5), control group for determination of the upper limit of autoregulation with stepwise induced arterial hypertension by intravenous administration of the alpha adrenergic drug methoxamine under continuous monitoring of mean arterial blood pressure (MABP); (2) Group B (n = 23), in which two cortical veins were photochemically occluded with rose bengal dye and fiberoptic illumination upon baseline CBF measurement. This was followed by repeated rCBF measurements under AR testing. Loss of AR in control Group A with passive increase of rCBF occurred at MABP of 147.5 +/- 2.9 mmHg. In Group B venous occlusion was followed by an initial phase of reduced rCBF, and then by pressure passive increases, thereby indicating loss of AR. Statistically significant changes of rCBF when compared to baseline MABP occurred at MABPbaseline + 10% (112.7 +/- 6.6 mmHg). We conclude that AR is impaired upon cortical venous occlusion with the propensity for hyperperfusion injury at a lower level of MABP when compared with a control group. In the context with earlier findings this may lead to narrowing of the corridor for MABP management following intra-operative occlusion of large cortical veins. PMID- 11877905 TI - Spontaneous recurrent seizures and neuropathology in the chronic phase of the pilocarpine and picrotoxin model epilepsy. AB - In a recent publication, we have shown a potent interaction between the cholinergic and GABAergic systems in regard to seizure generation and developed the pilocarpine(pilo)/picrotoxin(PTX) model, in which combined injections of these agents have induced status epilepticus (SE) in rats. Here we report on the chronic features of this new animal model of epilepsy. Adult male Wistar rats were systemically injected with solutions containing 150/0.5 mg kg-1, 75/1.5 mg kg-1 and 50/2.0 mg kg-1 (pilo dose/PTX dose). Six epileptic and six control animals were observed for 120-131 days for the occurrence of spontaneous recurrent seizures (SRS). Electroencephalographic, neuropathologic and behavioral analyses were subsequently performed. Following SE, the animals went through a latent period and, subsequently, towards a state of 'chronic' epilepsy, characterized by the emergence of SRS. Animals that received 150/0.5 mg kg-1 presented a relatively short latent period, partial events as their most common initial seizure manifestations and a considerable subsequent progression towards generalization. The group injected with 75/1.5 mg kg-1 presented an extensive period during which the majority of the animals exclusively developed partial seizures (50 days). Animals injected with 50/2.0 mg kg-1 presented an average latent period of over 100 days. Only few animals within this group developed SRS. Our EEG, neuropathological and ictal behavioral findings, in conjunction with the fact that SE was required for the posterior development of SRS, suggest that our model parallels a human TLE condition. Even though diverse TLE models have been described, the pilo/PTX model has as a major feature the intriguing occurrence of disparities among these three groups in the chronic period, although no differences could be observed during SE induction. Future experiments conducted in this sense, might lead to important results in regard to the elucidation of mechanisms of epileptogenesis. PMID- 11877908 TI - Contamination, or what is the price of the HIPAA? PMID- 11877907 TI - High-dose methylprednisolone prevents vasospasm after subarachnoid hemorrhage through inhibition of protein kinase C activation. AB - We have previously shown that the inflammatory process after subarachnoid hemorrhage causes vasospasm. The efficacy of methylprednisolone by suppression of the inflammatory process has been reported, although pharmacological mechanisms have not been clarified. The purpose of this study was to investigate the pharmacological mechanism of methylprednisolone on vasospasm. Using the 'two hemorrhage' canine model, progression of angiographic vasospasm was assessed in nontreated and treated groups with methylprednisolone. Methylprednisolone 10 mg kg-1 was injected i.v. after the first injection of blood, and the same dose was injected every 12 h until day 7. Protein kinase C (PKC) activity of canine basilar arteries in both groups was measured during the course of vasospasm. In the isometric tension study, the effect of methylprednisolone on tensions induced by phorbol 12-myristate 13-acetate (PMA), or high-K+ solution, was also evaluated. Methylprednisolone significantly reduced severity of vasospasm. In the treated group, PKC activity was not enhanced compared with the nontreated group at any point. Methylprednisolone inhibited tonic tension induced by PMA, but not that induced by high-K+ solution. We conclude that methylprednisolone prevents severity of vasospasm through inhibition of PKC activation, but does not work as a Ca2+ channel blocker. PMID- 11877909 TI - The first mortality follow-up study: the 1841 Report of William Farr (physician) on the mortality of lunatics. AB - BACKGROUND: In the 1830s in England, there was a great cultural interest in the collection and publishing of all kinds of statistics. The Council of the Statistical Society of London (founded in 1834) commissioned one of its Fellows, Dr William Farr, to investigate and prepare a report on the mortality of patients in the county asylums, with the mortality in a large number of proprietary houses that were licensed to care for patients with mental illness (then called lunatics) ordered for confinement because of their mental condition. Committees of Parliament had investigated the condition of the mentally ill confined to the asylums and taken measures in an attempt to improve their treatment and to correct abuses. RESULTS: Farr collected data from Hanwell, the Middlesex County asylum opened in 1831, and other asylums on annual admissions, resident patients, deaths, discharges, derived years of residence (exposure to risk), and annual mortality rates by duration and as an aggregate. He used similar data from a recent report on a large number of licensed houses. For the best estimate of comparative mortality, an assumed age distribution by sex and rates from the English Life Table No. 1 (constructed by Farr for 1841). CONCLUSION: Farr demonstrated that annual mortality rates were higher at durations 0-1.5 years than at durations 1.5-7.5 years, higher in men than in women, higher in paupers than in other patients, higher in licensed houses than in the Hanwell Asylum, and higher in the 4 large licensed houses than in a collection of smaller ones. COMMENT: A brief sketch of Farr's life is given as a memorial tribute to his pioneer work in vital statistics, life table methodology, public health, and life insurance medicine. PMID- 11877910 TI - Life table methodology applied to NHANES II database for analysis of mortality associated with cholesterol/HDL ratios. AB - This article describes the association between total cholesterol to high-density lipoprotein (HDL)-cholesterol ratios and all-cause mortality in a large cohort of Americans with nearly 17 years of follow-up. Detailed actuarial life table methodology was used. It concludes that the relationship is best described as a J shaped curve. PMID- 11877911 TI - Mortality outcomes after osteoporotic fractures in men and women. AB - BACKGROUND: Underwriting the elderly will challenge the skills of the medical director. Factors not typically reflecting an increased mortality risk in younger applicants assume major importance in the elderly. This article demonstrates osteoporotic fractures in the elderly can be predictive of adverse mortality. MATERIALS AND METHODS: In a 5-year prospective community study, all residents age 60 and over were screened for low-impact fractures, defined as those from a standing height or less. Two fracture groups were analyzed: proximal femur (hip) and combined vertebral and other major fractures. Those with predisposing underlying disease were excluded. Follow-up was nearly 100%. Age- and sex specific mortality for expected and those with each fracture group were calculated. Through an abridged life table analysis technique, the authors were able to create a 25-year cumulative survival analysis. RESULTS: There were more deaths among fracture patients in both groups than in the expected general population. Females with vertebral and other major fractures had a mortality ratio of 188% and excess death rate of 7. For hip fractures, values were 500% and 32. Males exhibited more adverse mortality, with a mortality ratio of 330% and excess death rate of 30 for vertebral and other major fractures and 540% and excess death rate of 57 for hip fractures. CONCLUSION: Osteoporotic fractures are risk factors for increased mortality in both males and females age 60 and older. The fractures contribute directly to mortality but more importantly appear to be a marker for comorbid conditions. PMID- 11877912 TI - Body mass index and mortality in an insured population. AB - OBJECTIVE: This study was conducted to explore the relationship between body mass index (BMI) and mortality in an insured population issued policies at standard rates or rated only for build using contemporary analytic techniques. BACKGROUND: Many factors influence the relationship between build and mortality. Recent clinical literature on this subject often employs multivariate statistical techniques to better define this relationship and reduce the influence of confounders. BMI, a common surrogate variable for build in clinical literature, is our variable of choice in studying the relationship between build and mortality. METHODS: We studied internal data on direct and reinsurance business issued between 1975 and 1998 at standard rates or rated only for build. The policies were followed till termination (death or lapse) or to the end of 1999. The average policy duration was 4.7 years. Cox proportional hazards model runs were used to study the multivariate relationship between mortality and BMI in moderately over- and underweight insured individuals. RESULTS: During follow-up, 4105 deaths were observed. Mortality was noted to very with BMI, most significantly in middle-aged male nonsmokers. Consistent with reports from the clinical literature, significant factors influencing the BMI-mortality relationship in this insured population included issue age and smoking status. CONCLUSIONS: BMI is a predictor of statistically significant mortality differentials in insured populations. The strength of the BMI-mortality relationship was found to vary by age, gender, and smoking status. In our study population, the male nonsmoker subgroups tended to exhibit the strongest graded relationship between hazard of death and increasing or decreasing BMI. PMID- 11877913 TI - Non-Hodgkin's lymphoma: in a class all its own. AB - Non-Hodgkin's lymphoma (NHL) incidence rates have increased more than 50% over the past 15 years, making it the sixth leading cause of death from cancer in the United States and the fourth most significant malignancy in terms of economic impact. Comparable trends are noted worldwide. Improved diagnostic techniques have resulted in reclassifying some tumors that in the past would have been classified as Hodgkin's disease. In this article, the latest diagnostic, prognostic, and treatment options for NHL are reviewed and a cross-reference chart for these evolving class systems provided. PMID- 11877914 TI - A quicker method for calculating mortality ratios based on survival rates in clinical trials and other follow-up studies. AB - OBJECTIVES: To provide reference expected cumulative survival tables, reducing steps in the calculation of mortality ratios and excess death rates from the medical literature, when source data are in terms of survival curves or cumulative survival rates. METHOD: Actuarial methodology. RESULTS: Tables provide cumulative expected survival rates calculated by entry age and for different periods of follow-up, as used in clinical trials and registries in the medical literature. CONCLUSION: The observed survival in scientific papers can be appraised promptly using cumulative survival tables calculated from either insurance or population life tables. PMID- 11877915 TI - Superior vena cava syndrome. AB - Approximately 15,000 cases of superior vena cava (SVC) obstruction are diagnosed in the United States annually. Malignancies (primarily lung cancer) are the underlying cause of 80-85% of cases, leaving 15-20% caused by various benign conditions, including sclerosing mediastinitis (the diagnosis in our case). Thrombolytic therapy and major advances in vascular techniques in recent years have improved the outcome and lessened the morbidity of SVC obstruction. However, even though a benign condition, sclerosing mediastitinis is a dynamic, ongoing fibrotic condition that seldom can be totally removed surgically. It frequently causes recurrent episodes of SVC obstruction, requiring further repetitive vascular procedures that can result in major morbidity and even mortality. PMID- 11877916 TI - Pulmonary flow loop. AB - Pulmonary flow loops display characteristic patterns and give clues to the underlying pathology of airway or parenchymal lung disease. PMID- 11877917 TI - Doppler echocardiographic changes found in diastolic dysfunction. AB - Diastolic dysfunction may be associated with increased morbidity and mortality. In this article, an overview of the pathophysiology of diastolic dysfunction is discussed and the findings of Doppler echocardiography used to aid in its diagnosis are reviewed. PMID- 11877918 TI - Underwriting chronic renal failure. AB - The progression of chronic renal failure is quite variable. Equations to estimate the level of function are provided. PMID- 11877919 TI - Staging and terminology of superficial urinary bladder tumors. AB - Ta and T1 transitional cell urinary bladder tumors are prone to recurrence in a large percentage of cases, but they uncommonly progress to invasive tumors. On the other hand, Tis bladder tumors progress to invasive tumors in a majority of cases. Even though all of these tumors are found in the superficial layers of the bladder wall, their natural histories are significantly different. In addition, new terminology has been developed for these lesions. PMID- 11877920 TI - The future of the pacemaker. PMID- 11877921 TI - Alternating narrow and wide complex tachycardia. PMID- 11877922 TI - Radiofrequency catheter ablation of an incessant ventricular tachycardia following valve surgery. AB - Sustained monomorphic ventricular tachycardia (VT) after valve surgery represents a clinical entity with different tachycardia mechanisms. This case report describes an incessant VT after tricuspid and aortic valve replacement that did not respond to antiarrhythmic drug treatment. The tachycardia exhibited VA block and a right bundle branch block pattern with left-axis deviation, suggesting ventricular excitation via the left posterior fascicle. The electrophysiological study was limited by the prosthetic tricuspid and aortic valve replacement, therefore a transseptal approach was necessary to obtain access to the ventricular myocardium. Radiofrequency catheter ablation was performed in the proximal left bundle or distal His region with termination of the incessant VT followed by complete AV block. After pacemaker implantation using a transvenous right atrial and an epicardial ventricular lead, no VT reoccurrence could be documented. PMID- 11877923 TI - Change in morphology of reentrant atrial arrhythmias without termination following radiofrequency catheter ablation. AB - A 60-year-old woman who had previously undergone an atrial septal defect repair and had type I atrial flutter underwent electrophysiological study. After radiofrequency (RF) ablation to the isthmus between the inferior vena cava and the tricuspid annulus, type I atrial flutter was changed to atrial tachycardia following atriotomy without termination. This atrial tachycardia was eliminated by single-site RF ablation of a small lesion below the caudal end of the atriotomy scar, where continuous and fragmented potentials were recorded during tachycardia. We experienced a rare case in which RF energy changed tachycardia circuits. PMID- 11877924 TI - Rate dependent far-field R wave sensing in an atrial tachyarrhythmia therapy device. AB - A case is reported of far-field R wave (FFRW) oversensing resulting in inappropriate atrial tachycardia (AT) detection by a dual chamber pacemaker incorporating atrial autoadjusting sensitivity (AAS). FFRW oversensing occurred during periods of functional atrial undersensing (FAU) with PR interval prolongation. Limitations of the pacemaker's ability to reject FFRWs and programming considerations for addressing this unique behavior are discussed. PMID- 11877925 TI - Inappropriate mode switching in a dual chamber pacemaker due to oversensing of a high frequency signal from a conductor/ring discontinuity (loose set screw). AB - Two unusual cases are reported of pacemaker oversensing of the impedance pulse by the atrial channel attributed to a loose set screw. PMID- 11877926 TI - Radiofrequency catheter ablation of a right free-wall accessory pathway in dextrocardia with complete situs inversus. AB - An uncommon case of orthodromic atrioventricular reentrant tachycardia in a patient with dextrocardia and complete situs inversus is reported. There was no associated cardiac abnormality apart from the mirror image dextrocardia. An electrophysiological study and successful catheter ablation of a right free-wall concealed accessory pathway was performed. A simplified three-catheter technique from a femoral approach and monoplane fluoroscopy were used in a noncomplicated procedure of normal duration. PMID- 11877927 TI - Neurocardiogenic syncope due to recurrent tonsillar carcinoma: successful treatment by dual chamber cardiac pacing with rate hysteresis. AB - A patient with recurrent tonsillar carcinoma in his neck presented with cardioinhibitory neurocardiogenic syncope. Ventricular demand pacing was ineffective in preventing recurrent symptoms. The pacemaker was upgraded to a dual chamber device with hysteresis that eliminated further syncopal episodes. PMID- 11877928 TI - Inappropriate tachycardia detection by a biventricular implantable cardioverter defibrillator. AB - This case describes "double counting" of intrinsic conduction due to the combination of intraventricular conduction delay and the merging of the electrograms from the right and left ventricle (coronary sinus) leads in a biventricular cardioverter defibrillator. This situation prompted asymptomatic antitachycardia pacing. Lengthening of AV conduction by betablocker solved the problem. PMID- 11877929 TI - Advisory information from Vitatron. PMID- 11877931 TI - Computerized cardiac mapping. PMID- 11877930 TI - Effect of ventricular fibrillation duration on the defibrillation threshold in humans. AB - Early during ventricular fibrillation, the defibrillation threshold may be low, as ventricular fibrillation most probably arises from a localized area with only a few wavefronts and the effects of global ischemia, ventricular dilatation, and sympathetic discharge have not yet fully developed. The purpose of this study was to explore the effect of the timing of shock delivery in humans. During implantation of an ICD in 26 patients (24 men, 60 +/- 11 years, 19 coronary artery disease, NYHA 2.2 +/- 0.4, left ventricular ejection fraction 0.42 +/- 0.16), the defibrillation threshold was determined after approximately 10 and 2 seconds of ventricular fibrillation. Ventricular fibrillation was induced by T wave shocks. Mean defibrillation threshold was 9.9 +/- 3.6 J after 10.3 +/- 1.0 seconds. Within 2 seconds, 20 of 26 patients could be successfully defibrillated with < or = 8 J. In these patients, the mean defibrillation threshold was 4.0 +/- 2.1 J after 1.4 +/- 0.3 seconds compared to 9.5 +/- 3.1 J after 10.2 +/- 1.1 seconds (P < 0.001). There were no clinical differences between patients who could be successfully defibrillated within 2 seconds and those patients without successful defibrillation within 2 seconds. In the majority of patients, the defibrillation threshold was significantly lower within the first few cycles of ventricular fibrillation than after 10 seconds of ventricular fibrillation. These results should lead to exploration of earlier shock delivery in implantable devices. This could possibly reduce the incidence of syncope in patients with rapid ventricular tachyarrhythmias and ICDs. PMID- 11877932 TI - QT dispersion in 120 electrocardiographic leads in patients with structural heart disease. AB - The clinical significance of QT dispersion (QTd) measured in 12-lead ECGs is controversial. The aim of this study was to clarify factors that determine the QTd and its measurement errors in different lead arrays in patients with structural heart disease. Two blinded observers measured QT intervals on a computer screen from 120-channel ECG recordings in a retrospective set of 257 patients, comprising a group of 121 myocardial infarction (MI) survivors without ventricular tachyarrhythmia during a 6-month follow-up and a group of 136 survivors of ventricular tachyarrhythmia/fibrillation. QTd did not differ in patients with and without ventricular tachyarrhythmia/fibrillation. Eleven ventricular tachyarrhythmia/fibrillation survivors without structural heart disease had the lowest QTd (P < or = 0.02). The strongest factor determining QTd and the magnitude of its measurement error was the lead array (P = 0.0001). Measurement errors had two components. The smallest relative errors were in the total body surface mapping array with one component related to interobserver reproducibility (9.1 +/- 7.6%), and the other component related to accuracy of measurement of the QT interval (36 +/- 16%). The authors estimated that a difference of QTd of at least 50 ms between study groups is required in a 12-lead ECG to draw any conclusions from the studies. In patients with structural heart disease, QTd from limited arrays of ECG leads was not a reliable measure. It correlated with the presence of structural heart disease, but not with arrhythmogenicity. An array consisting of ECG leads covering the entire chest allowed better reproducibility and measurement accuracy of QTd. PMID- 11877933 TI - Fiberoptic balloon catheter ablation of pulmonary vein ostia in pigs using photonic energy delivery with Diode laser. AB - Circumferential lesions to the pulmonary vein (PV) ostia to cause conduction block at the junction of the PV and left atrium could offer a new approach during catheter ablation of patients with paroxysmal (focal) atrial fibrillation. Diode laser can deliver energy through diffusing or ring fiber tips. In three pigs weighing between 60 and 65 kg, transseptal puncture was performed and a fiberoptic balloon catheter with a collapsed profile of 10 Fr was advanced through a sheath under fluoroscopic guidance to the ostium of the right and left PVs. The balloon was inflated with a 3-cc mixture of D2O (deuterium oxide) and contrast to deliver circumferential lesions with a 15-mm diameter x 3-mm ring width of light. Applications consisted of 3.2 to 3.8 W/cm for 120 seconds; the animals were sacrificed 3 hours after ablation for pathological examination. Photonic energy was delivered successfully to the ostium of five of the five targeted PVs, and was well tolerated hemodynamically in each animal without ectopy. Gross inspection revealed endocardial lesions at the ostium of four of five PVs, confined to the atrium in each and circumferential in three of five PVs. Microscopically, transmural coagulation necrosis of the atrium was present at the ostium of three of five PVs, and extended into the myocardial sleeves of two PVs. Photonic energy delivery using a fiberoptic balloon catheter can create circumferential lesions to the PV ostia, suggesting that this new form of energy delivery may be therapeutically advantageous for pulmonary vein ablation with need to pursue chronic studies. PMID- 11877934 TI - Predicting the outcome of patients with unexplained syncope undergoing prolonged monitoring. AB - Patients with unexplained syncope are often considered candidates for prolonged monitoring or empiric pacing when noninvasive and invasive investigations fail to provide a diagnosis. Identifying the outcome of patients undergoing prolonged monitoring that would ultimately benefit from empiric pacing may permit a cost effective approach to resolution of syncope. Two hundred and six patients (age 57 +/- 18 years, 57% male) underwent prolonged monitoring with an implanted loop recorder for syncope of unknown origin. The median number of previous syncopal episodes was four (mean 29 +/- 133). Prior tilt testing was performed in 63% of patients, and electrophysiological testing in 46%. Symptoms recurred during follow-up in 142 patients (69%). Recurrence was associated with bradycardia leading to pacemaker implantation in 35 patients (17.0%), tachycardia in 12 (5.8%), sinus rhythm in 63 (30.6%), neurally mediated syncope based on rhythm and clinical assessment in 22 (11%), and failed activation in 10 (5%). Logistic regression analysis of baseline variables found that age was the only independent variable that predicted the need for pacing, associated with a 3% increase in risk per advancing year of age (odds ratio 1.027, P = 0.026). Despite this finding, no age group could be identified in which the likelihood of requiring pacing exceeded 30%. Logistic regression also found that patients with structural heart disease were less likely to experience recurrent symptoms during monitoring (49% vs 78%, P = 0.001) and that advancing age was associated with earlier recurrence of symptoms (P = 0.01). The etiology of recurrent syncope is diverse and cannot be predicted by baseline clinical variables. Empiric pacing appears to have little role in the management of this patient population. PMID- 11877935 TI - The effect of ablation electrode length and catheter tip to endocardial orientation on radiofrequency lesion size in the canine right atrium. AB - Although the determinants of radiofrequency lesion size have been characterized in vitro and in ventricular tissue in situ, the effects of catheter tip length and endocardial surface orientation on lesion generation in atrial tissue have not been studied. Therefore, the dimensions of radiofrequency lesions produced with 4-, 6-, 8-, 10-, and 12-mm distal electrode lengths were characterized in 26 closed-chested dogs. The impact of parallel versus perpendicular catheter tip/endocardial surface orientation, established by biplane fluoroscopy and/or intracardiac echocardiography, on lesion dimensions was also assessed. Radiofrequency voltage was titrated to maintain a steady catheter tip temperature of 75 degrees C for 60 seconds. With a perpendicular catheter tip/tissue orientation, the lesion area increased from 29 +/- 7 mm2 with a 4-mm tip to 42 +/ 12 mm2 with the 10-mm tip, but decreased to 29 +/- 8 mm2 with ablation via a 12 mm tip. With a parallel distal tip/endocardial surface orientation, lesion areas were significantly greater: 54 +/- 22 mm2 with a 4-mm tip, 96 +/- 28 mm2 with a 10-mm tip and 68 +/- 24 mm2 with a 12-mm tip (all P < 0.001 vs perpendicular orientation). Lesion lengths and apparent volumes were larger with parallel, compared to perpendicular tip/tissue orientations, although lesion depth was independent of catheter tip length with both catheter tip/tissue orientations. Electrode edge effects were not observed with any tip length. Direct visualization using intracardiac ultrasound guidance was subjectively helpful in insuring an appropriate catheter tip/tissue interface needed to maximize lesion size. Although atrial lesion size is critically dependent on catheter tip length, it is more influenced by the catheter orientation to the endocardial surface. This information may also be helpful in designing electrode arrays for the creation of continuous linear lesions for the elimination of complex atrial tachyarrhythmias. PMID- 11877936 TI - Effect of electrode location in great cardiac vein on the ventricular defibrillation threshold. AB - This study tested the hypothesis that the DFT could be lowered by delivering a weak auxiliary shock in conjunction with a stronger primary shock with the auxiliary shock electrode near the cardiac region where the primary shock electric field is weakest. This hypothesis was tested by determining the DFTs with the auxiliary shock delivered from different locations within the great cardiac vein (GCV). In 15 dogs, catheters with defibrillation electrodes were placed transvenously in the RV apex, the SVC, and the GCV. An active can electrode and the SVC electrodes were electrically coupled to serve as a return electrode for the RV and GCV electrodes. DFTs were determined for a primary shock through the RV electrode with and without a subsequent auxiliary shock of lower amplitude through the GCV electrode. The leading edge voltage and current at DFT were significantly lowered by addition of the auxiliary shock (17% and 19% decreased, respectively), but energy was not changed. The animals were divided into three groups according to the location of the GCV electrode. The leading edge voltage, current, and total delivered energy at the DFT were significantly lower in animals with the GCV electrode near the apex (22%, 24%, and 13% reduction, respectively) compared with those where the GCV electrode was positioned away from apex (8%, 10% reduction and 18% increase, respectively, P < 0.001). Application of an auxiliary shock to the apical region, near the region where previous studies have indicated that the RV primary shock has its weakest effects, caused the greatest decrease in DFT. PMID- 11877937 TI - Clinical experience with electroanatomic mapping of ectopic atrial tachycardia. AB - The aim of this study was to evaluate the clinical use of a new three-dimensional mapping system as a guide for catheter ablation of ectopic atrial tachycardia. A series of 42 consecutive patients with drug refractory ectopic atrial tachycardia was studied in a prospective observational trial with the electroanatomic mapping system CARTO. The arrhythmogenic focus was found in the right atrium in 30 patients and in the left atrium in 12 patients. The construction of a complete electroanatomic map of the right or left atrium was possible in 37 of 42 consecutive patients with ectopic atrial tachycardia. Mean activation time of the right atrium, including the proximal coronary sinus, was 94 +/- 25 ms for right atrial tachycardias; left atrial activation time during left atrial tachycardias was 86 +/- 17 ms. Average mapping time was 30 minutes for right atrial tachycardias and 22 minutes for left atrial tachycardias, allowing the collection of 86 +/- 50 and 65 +/- 28 catheter positions, respectively. The size of the area of earliest atrial activation calculated from the electroanatomic map amounted to 0.6 +/- 0.4 cm2 in right atrial tachycardias and 1.0 +/- 0.9 cm2 in left atrial tachycardias. In the right atrium the most common locations of the 33 arrhythmogenic foci in 30 patients were the high or mid-lateral right atrium (n = 10) and the inferoparaseptal region near the coronary sinus ostium (n = 7). Ectopic left atrial foci were most commonly located in an inferior position near the mitral annulus (n = 5) and in proximity to the ostium of the pulmonary veins (n = 4). Biatrial electroanatomic mapping allowed visualization of earliest right atrial activation during left atrial tachycardia at the high interatrial septum or near the coronary sinus ostium. Catheter ablation was successful in 85% of right atrial tachycardias and 82% of left atrial tachycardias. In patients with ectopic atrial tachycardia electroanatomic mapping is a safe and feasible technique that allows three-dimensional visualization of the automatic focus in a precise anatomic reconstruction of the atria. This novel mapping technology facilitates catheter ablation of complex ectopic atrial tachycardia. PMID- 11877938 TI - Nonfluoroscopic magnetic electroanatomic mapping to facilitate focal pulmonary veins ablation for paroxysmal atrial fibrillation. AB - RF ablation of ectopic foci in the pulmonary veins (PVs) is a promising treatment for patients with paroxysmal AF. The aim of this study was to evaluate the feasibility of using nonfluoroscopic magnetic electroanatomic mapping of PV during spontaneous or induced ectopy to facilitate focal ablation procedure. The study included 35 patients with drug refractory paroxysmal AF who underwent focal RF ablation of the PV. In 10 (29%) patients, mapping and RF ablation procedures were performed using the nonfluoroscopic magnetic electroanatomic mapping system to enable automatic capture of the location and the timing of the ectopy. As a control, 25 patients underwent conventional endocardial activation mapping technique. There were no significant differences in the clinical characteristics between the two groups. Overall procedural duration was similar between them (199 +/- 52 vs 221 +/- 82 minutes, P > 0.05). However, the mean fluoroscopy time (25 +/- 6 vs 52 +/- 12 minutes, P = 0.01) and the mean number of RF applications (5 +/- 3 vs 12 +/- 9, P = 0.02) were significantly less in patients who underwent electroanatomic mapping. There were no significant differences between the two groups in the acute (90 vs 84%) and long-term success rate (60 vs 56%) after a mean follow-up of 12 +/- 9 months. In conclusion, RF ablation of ectopic foci using nonfluoroscopic magnetic electroanatomic mapping of PVs during spontaneous or induced ectopy is useful even in patients with a limited number of ectopy, and is associated with a similar success rate, but less fluoroscopy time and RF application compared to the conventional approach. PMID- 11877939 TI - Timing cycles for biventricular pacing. PMID- 11877940 TI - Fluoroscopic cardiac anatomy for catheter ablation of tachycardia. AB - The understanding of cardiac anatomy is crucial for the interventional arrhythmologist. In spite of the introduction of several nonfluroscopic navigational tools, some of them capable of reconstructing a computer-based surrogate of the endocardial surface of the heart cavities, simple fluoroscopy with or without the aid of angiographic techniques is still the most widely used method to guide mapping and ablation procedures. In some instances, fluoroscopic and angiographic methods have no possible replacement to unravel certain arrhythmologically useful anatomic landmarks. New interpretations of cardiac architecture show the need to challenge some traditional anatomic views, like the concept of septums within the heart. The fluoroscopic anatomy also needs to be reconsidered in the light of the new attitudinally oriented nomenclature. This article presents an overview of the fluoroscopic anatomy of the heart. When pertinent, some anatomical concepts are discussed in more detail like the triangle of Koch, the pyramidal space, and the interatrial groove. In the sections on the atria and on the ventricles, the authors focus on the anatomic information that is relevant for mapping and ablation from a fluoroscopic viewpoint, providing some hints on how best to depict the morphological features from the stance of the interventional arrhythmologist. The Visible Human Slice and Surface Server using data sets from the Visible Human Male and Female Project, has been used to facilitate the understanding of the fluoroscopic anatomy. PMID- 11877941 TI - A comparison of left atrial size by two-dimensional transthoracic echocardiography and magnetic endocardial catheter mapping. AB - This study sought to validate the accuracy of magnetic electroanatomic mapping (MEAM) for determining cardiac chamber size in a clinically relevant situation. The authors chose to compare LA size measured by MEAM to that assessed by two dimensional guided M-mode echocardiography. The study included 37 patients with drug refractory paroxysmal atrial fibrillation who underwent two-dimensional echocardiography and a detailed MEAM of the LA. The entire LA was mapped with a mean of 132 +/- 50 points with attention to identifying the mitral annulus and posterior wall of the LA. The MEAM measurement of LA size was taken as the distance from the anterior wall of the LA to the posterior wall in a plane parallel to the mitral valve annulus at atrial end-diastole. LA dimension determined by M-mode echocardiography was assessed in a plane parallel to the mitral valve annulus in the parasternal long-axis view during atrial end diastole. LA size assessed by M-mode echocardiography was 41.2 +/- 5.0 versus 40.9 +/- 4.5 mm as assessed by MEAM, with good correlation (r = 0.87, P < 0.001). Only three patients had a difference in LA size that was > 0.3 cm between the two measurement techniques. Thus, it appears that LA anteroposterior dimension as determined by electroanatomic mapping is similar to that determined by two dimensional guided M-mode echocardiography. MEAM appears to be an accurate method by which LA size can be assessed in patients with drug refractory atrial fibrillation undergoing left atrial ablation procedures. PMID- 11877942 TI - Adenosine triphosphate in cardiac arrhythmias: from therapeutic to diagnostic use. PMID- 11877944 TI - North Share-Long Island Jewish invests in lifelong learning program. PMID- 11877943 TI - Ambulatory care: what's profitable, what's not. PMID- 11877945 TI - VHA develops safety program to prevent surgical site errors. PMID- 11877946 TI - Leapfrog's report is incomplete, misleading. PMID- 11877947 TI - Harris Poll shows most people like their health plans, if not managed care in general. PMID- 11877948 TI - [Treatment of chronic hepatitis B]. PMID- 11877949 TI - [Antiviral therapy for liver cirrhosis type B]. PMID- 11877950 TI - [Clinical study of dissection of the superior mesenteric artery]. AB - Six patients with dissection of the superior mesenteric artery (SMA) who were treated at our hospital between 1993 and 1999 were studied. Hypertension was considered significant as a risk factor. The clinical features were characterized by severe abdominal pain which radiated posteriorly, back pain, weak bowel sounds and exacerbation by eating. An ultrasonographic examination and a computed tomographic scan of the abdomen were useful for diagnosis. Four cases showed full recovery under conservative management with anticoagulant or antiplatelet drugs. Two cases without recovery should be considered intervention if they have progressive courses. This disease should be kept in mind on differential diagnosis of abdominal pain, because it may be overlooked without a correct diagnosis. PMID- 11877951 TI - [A case of Crohn's disease complicated by liver abscess]. PMID- 11877952 TI - [Six cases of incarcerated obturator hernia preoperatively diagnosed by pelvic CT]. PMID- 11877953 TI - [A case of epilepsy after therapy with interferon-alpha-treated chronic hepatitis C]. PMID- 11877954 TI - [A case of toxic shock syndrome and acute cholecystitis caused by MRSA]. PMID- 11877956 TI - [A case of intraductal adenocarcinoma of the pancreas with Peutz-Jeghers syndrome]. PMID- 11877955 TI - [A case of primary sclerosing cholangitis presenting transient hypoperfusion and treated with bezafibrate beneficially]. PMID- 11877957 TI - [Management of patients with bronchial asthma received general anesthesia and surgical intervention]. AB - Stimulation to bronchial mucosa is one of the major risk factor of asthma attack. When patients receive surgical intervention and general anesthesia, they are always exposed to stimulation to bronchial mucosa. Prevention method of bronchial asthma attack during surgical intervention is not established yet. We investigated that clinical course of patients with bronchial asthma who received general anesthesia and surgical intervention. Seventy-six patients with bronchial asthma were received general anesthesia and surgical intervention from 1993 to 1998. Twenty-four patients were mild asthmatic patients, 39 were moderate asthmatic patients and 13 were severe asthmatic patients. Preoperative treatment for preventing asthma attack was as follows; Eight patients were given intravenous infusion of aminophylline before operation. Fifty-two patients were given intravenous infusion of aminophylline and hydrocortisone before operation. Three patients were given intravenous infusion of hydrocortisone for consecutive 3 days before operation. Thirteen patients were given no treatment for preventing asthma attack. One patient was suffered from asthma attack during operation. She was given no preventing treatment for asthma attack before operation. Three patients were suffered from asthma attack after operation. No wound dehiscence was observed in all patients. To prevent asthma attack during operation, intravenous infusion of steroid before operation is recommended, when patients with asthma receive general anesthesia and surgical intervention. PMID- 11877959 TI - [Evaluation of the enzyme-linked immunosorbent assay system for sero-diagnosis of summer-type hypersensitivity pneumonitis]. AB - Summer-type hypersensitivity pneumonitis (SHP) is the most prevalent type of hypersensitivity pneumonitis in Japan. Its main causative antigenic organisms are Trichosporon species. Although this disease had been thought to be unique to Japan, more than ten patients were found in Korea, recently. It is expected that patients with SHP will be found elsewhere in the Temperate Zones, the tropics or in the subtropics. It is important in diagnosing SHP to prove specific antibody against causative antigen in patient's serum, for the existence of specific antibody indicates the patient had been exposed to, and sensitized to the antigen. In the present study, we devised Enzyme-linked Immunosorbent Assay system using monoclonal antibody specific to T. asahii, to measure antibodies against T. asahii. We evaluated the usefulness of the system by using sera from patients with SHP, patients with other pulmonary diseases, or healthy volunteers. We found specificity and sensitivity of the system were 92.8% and 92.3%, respectively. This system was proved to be useful in diagnosing SHP. PMID- 11877958 TI - [Cedar pollinosis in Japanese schoolchildren: results from a large questionnaire based survey]. AB - PURPOSE: To evaluate the prevalence and clinical characteristics of Japanese cedar (Cryptomeria japonica) pollinosis (CP) in Japanese schoolchildren. METHODS: Questionnaire made by the Study Group of Epidemiology of Allergic Diseases founded by the Japanese Ministry of Public Health and Welfare was distributed to 56,108 schoolchildren living in the suburban areas of Kyoto, Japan. The data recovered from 50,086 children (recovery rate 89.3%) was analyzed. RESULTS AND DISCUSSION: Overall prevalence of CP was 5.2%. The prevalence was higher in older children, and in those born in autumn and winter. Prevalence of CP in southern urban area was significantly higher (p < 0.001) than that in northern rural area, suggesting that urban environment including air pollution might increase the prevalence of CP. Among children with AD, there was statistically significant correlation between the severity of AD and the presence of CP (p = 0.016), i.e. those with CP tended to have more severe AD symptoms. On the other hand, the severity of BA was not affected by the presence of CP (p = 0.323). These data suggest a contribution of cedar pollen to AD, but not to BA, symptoms. Overall, our present data shown that CP has become one of the important allergic diseases in childhood, and that further evaluation of CP among children would be necessary. PMID- 11877961 TI - [An investigation of the use of diskhaleres in asthmatic patients]. PMID- 11877960 TI - [Real-time measurement of airborne pollen allergen applied the surface plasmon resonance (SPR)]. AB - To examine the possibility of real-time measurement system for the concentration of airborne pollen allergens, Cry j 1 was chosen as an example to establish the system. Airborne pollen allergens were collected using Cyclone sampler from Burkard Co. Ltd. England. Cry j 1 was extracted from airborne samples with 10 mM HEPES buffer containing 0.125 M NH4HCO3 and measured with Biacore 3000 and Biacore Q. The sensitivity of the system was 5 ng/ml, and 0.1 ml sample volume and at least 500 pg of Cry j 1 was required for each measurement. Amounts of Cry j 1 can be obtained ca. 40 min after collection, that is, 15 min for extraction, 10 min for separation and 10 min for measurement. PMID- 11877962 TI - [Gramineae pollen dispersal and pollinosis in the city of Hisai in Mie Prefecture. A 14-year study of gramineae pollen dispersal and cases of sensitization to gramineae experienced at an allergy clinic over a 15-year period]. AB - (1) A study of Gramineae pollen dispersal was conducted in Hisai, Mie Prefecture from 1987 to 2000; Gramineae pollen counts did not increase, with an average of 133 during the 14 years. There was two or three peaks in the annual dispersal period, suggesting that symptoms in late May, late July to early August, and late September are important for diagnosis of Gramineae pollinosis in this region. (2) Changes in the frequencies of sensitization to Gramineae pollen and Gramineae pollinosis were studied in 756 allergy clinic patients with nasal allergies examined over a 15-year period from 1986 to 2000. Results for the frequency of sensitization every 5 years came to 7.9, 22.4, and 23.3% and that of Gramineae pollinosis were 4.8, 10.3, and 12.1%. Both frequencies tended to increase after 1990. Out of 135 patients with sensitization to Gramineae, 88.1% had multiple sensitization to Japanese cedar and 54.8% had sensitization to Japanese cedar as well as house dust or mites. Patients sensitized only to Gramineae accounted for 5.9% of total patients. About half of the patients with sensitization to Gramineae developed pollinosis. As the CAP RAST score increased, so did the increase of pollinosis. PMID- 11877963 TI - [Lemofloxacin: antimicrobial ability and clinico-pharmacokinetic basis for use in urogenital infections]. AB - Lomefloxacin, as other fluoroquinolones, is a drug of choice in the treatment of uncomplicated urinary infections (acute cystitis, pyclonephritis) in outpatient practice. The advantage of lomefloxacin consists in a single-dose regimen (400 mg with 24-h interval). A course of acute cystitis lasts 3 days, acute pyelonephritis--10-14 days. Lomefloxacin is also the first-line drug in exacerbation of mild or moderate chronic pyclonephritis in hospitals and outpatient clinics. Fluoroquinolones are now most effective in management of acute or exacerbation of chronic bacterial prostatitis. Optimal lomefloxacin regimen in prostatitis is 400 mg with 24-h interval for one month. For prevention of postoperative suppuration in prostatic resection lomefloxacin is given in a single dose 400 mg 3-6 hours prior to operation. If operation is to be performed in the presence of urinary infection or prostatitis, lomefloxacin should be taken for 5-7 days before surgery. PMID- 11877964 TI - [Comparative analysis of long-term results of treating chronic prostatis with the use of the Andro-Gin device]. AB - Follow-up examinations covered 91 patients aged from 20 to 60 years with chronic prostatitis (CP) history 1-18 years. The primary examination has found that 79.1% examinees has CP complicated with sexual dysfunction, disturbed spermatogenesis and psychoneurological disorders. In one group of CP patients etiopathogenetic treatment of CP was combined with magneto-laser-electrostimulation of the prostate provided by the unit Andro-Gin. This combined treatment proved more effective as it induced long-term remission (two years and longer) in 60.5% patients. Unsatisfactory results were minimal. Without use of the unit unsatisfactory results were recorded in 35% patients. PMID- 11877965 TI - [Immunologic assessment of the risk of developing infectious-inflammatory complications after percutaneous operations for nephrolithiasis]. AB - A retrospective comparison of the evidence obtained at preoperative examination of 68 patients with urolithiasis operated with the use of percutaneous technologies has demonstrated that the risk of postoperative infectious inflammatory complications depends much on the immune status of the patient. Patients with initially different states of phagocytic immunity and antibody production had different courses of the postoperative period. An algorithm of immunological prediction of an acute pyelonephritis attack after percutaneous operations for nephrolithiasis is proposed. PMID- 11877966 TI - [Comparative evaluation of the effectiveness and safety of combined drug therapy of patients with benign prostatic hyperplasia with finasteride and alfuzozine]. AB - A finasteride + alfuzozine combination was used in 3-year treatment of 138 patients with initial benign prostatic hyperplasia (BPH). The principle of the combination is in parallel administration of 5 alpha-reductase inhibitors which inhibit cell proliferation at the hormonal level and alpha-adrenoblockers affecting the smooth muscle component of prostatic stroma and detruzor blood supply. Patients participating in the study had enlanged prostate (at least 60 cm3) and pronounced symptoms (IPSS over 13 scores). Patients of two control groups received alfuzozine and finasteride monotherapy, respectively. Better micturition and relief of BPH symptoms were seen in 96% patients of the study group, 84 and 74% controls, respectively. Thus, compared to monotherapy, finasteride and alfuzozine were more effective in combination which is pathogenetically valid and perspective in BPH chemotherapy. PMID- 11877968 TI - [Fluorescent cystoscopy in diagnosing and treating superficial cancer of the urinary bladder]. PMID- 11877967 TI - [Five-year experience in treating patients with prostatic hyperplasia patients with permixone (Serenoa repens "Pierre Fabre Medicament)]. AB - Specialists of the urologic clinic of the I.M. Sechenov Moscow Medical Academy studied effectiveness of lipidosterol extract Serenoa repens (permixon) in 26 patients with prostatic hyperplasia (total prostate-specific antigen was under 4 ng/ml). The trial has been performed from November 1995 up to now. The drug was taken before meal with a small quantity of water in a total daily dose 320 mg twice a day. Initial IPSS values ranged from 8 to 18 scores (mean 11.65 +/- 0.59). Life quality index was 1 to 4 scores (mean 2.46 +/- 0.15). Initial size of the prostate varied from 26 to 63 cm3 (mean 36.23 +/- 1.57 cm3). Maximal urinary flow rate (Qmax) made up 8.7 to 14.6 ml/s (mean 11.83 +/- 0.31 ml/s). Residual urine was initially 0-60 ml (mean 10.58 +/- 2.91 ml). Permixon significantly reduced the disease symptoms and improved quality of life. 5 years of treatment decreased mean IPSS by 8.8 +/- 0.18 (75.5%). QOL--by 1.31 +/- 0.08 (53.3%), size of the prostate--by 10.81 +/- 0.55 cm3 (29.8%). Neither the symptoms nor quality of life became worse for these five years. The size of the prostate reduced in 16, unchanged in 9 and increased only in 1 patient. Qmax was initially under 15 ml/s and rose after the treatment by 4.13 +/- 0.51 ml/s (35%), on the average. Qmax rose above 15 ml/s in 16 patients. Residual urine increased during the treatment in one patient only. Permixon intolerance was not observed. Thus, continuous 5-year therapy with lipidosterol extract Serenoa repens (permixon) proved highly effective and safe in 26 patients with initial or moderate symptoms of prostatic hyperplasia. PMID- 11877969 TI - [Long-term results of organ-preserving surgeries for renal cell carcinoma]. AB - The paper presents long-term results of conservative surgical treatment of 148 patients with renal cell carcinoma (RCC). Cancer-specific survival, local recurrence and distant metastases rates were estimated. Overall cancer-specific survival reached 91.8%. For patients with imperative indications this survival was 79.6%, for elective patients--98%. Recurrent RCC was observed in 10(6.8%) patients: imperative group--8(5.4%), elective group--2(1.3%). Distant metastases were detected in 7(4.7%) patients. The elective group had no metastases. General level of multifocality of sporadic renal cancer was 6.7%, for tumors < 4 cm- 1.2%. The basic criterion of the decision on organ-saving operation is tumor size under 4 cm. The size of the primary tumor determines the level of local recurrences and distant metastases. To select the optimal group of patients, genetic markers of tumor multifocality should be identified. PMID- 11877971 TI - [Endoluminal ultrasonography in diagnosing and treating upper urinary tract diseases]. AB - Endoluminal ultrasound scanning (EUS) of the upper urinary tracts (UUT) with intracavitary catheter sensor was performed in 15 patients (7 men and 8 women) aged 21 to 61 years. In 8 patients EUS was diagnostic (in one patient it was made twice, in 2 cases--in combination with ureteroscopy and biopsy), in 7 patients it was a diagnostic stage of the following surgical intervention, in 6--in combination with roentgenoendoscopic correction of strictures of the ureteropelvic segment (UPS) and in 1--in combination with transurethral pyelolithotripsy. A total of 16 procedures were made: 15 transurethral and 1- percutaneous. Endoluminal ultrasonography was conducted not only of the whole ureter but also of renal calycopelvic system (RAPS) which presented endosonographic semiotics of UUT. EUS has diagnosed urate concrement in 4 cases, "cross" vessels which narrowed UPS in 3 cases with late hydronephrotic transformation. In one patient EUS was carried out in combination with ureteroscopy and biopsy as a control examination 6 and 18 months after transurethral endoresection of ureteral tumor. In 6 patients with UPS strictures EUS was made for diagnosis of suspected cross vessels before endopyelotomy. No complications during EUS were encountered. Pilot experience with intracavitary ultrasonography of the UUT shows its high diagnostic potential and clinical value. This technique details inner structures of the ureter and RAPS with adjacent tissues due to minimal distance between the sensor and studied object. PMID- 11877972 TI - [Methods of diagnosing adenoma and hyperplasia of the parathyroid gland]. PMID- 11877973 TI - [Ways of improving preparing research and teaching personnel and clinicians in urology in the Russian Federation]. AB - The article covers basic principles of urologists' and researchers' training for state healthcare system. The role of a student research group in proper students selection for further special education in urology is pointed out. The article contains a plan of training in clinical residency including theoretical basis of urology and allied fields, diagnostic tools and practical skills, especially in urgent cases. The principles of postgraduate research personnel training in the field of urology are highlighted also. Special attention is paid to regional specialists training. The residents in surgery are preferred for further training in urology. PMID- 11877974 TI - [Temporary ureteral stents in patients with neuorgenic dysfunction of the urinary bladder]. AB - Biodegradable spiral stents were used in the treatment of detrusor external sphincter dyssynergia (4 patients) and detrusor areflexia with external sphincter spasticity (3 patients). All these patients had spinal injuries. After the stent placement all the patients could urinate almost without residual urine. In patients with detrusor-external sphincter dyssynergia mean voiding pressure was 79 cm water before treatment and 37 cm 10 weeks after the stent introduction. Biodegradable spiral stents are a promising treatment for patients with neurogenic bladder. PMID- 11877975 TI - [Operative treatment of complications caused by intracavernous administration of "Interfal" gel]. AB - The authors have operated 5 men for complications after introduction of gel "interfal" into cavernous bodies of the penis. The removal of the gel was accompanied with incomplete intracavernous prosthesis of the penis by M. N. Zilberman. A satisfactory functional result was achieved in 3 patients. Postoperative purulent cavernitis required removal of the prostheses in 2 men. Individual approaches to the treatment are recommended. The use of gel "interfal" must not be permitted for treatment of erectile dysfunction. PMID- 11877976 TI - [Pathogenetic bases for choosing treatment for vesicoureteral reflux in children]. AB - 600 children treated surgically or conservatively for vesico-ureteral reflux (VUR) at the age 3 months to 15 years were followed up for maximum 28 years. The course of compensatory processes in the kidney with considering the cause of VUR development at different stages of life (childhood, adolescence, adult life) and the degree of renal function loss were specified. Morphologic studies of renal parenchyma help choosing treatment of VUR in children depending on the cause of its development: malformation of ostium ureteris and intramural ureter, inflammation in the urinary bladder. PMID- 11877977 TI - [Primary transitional cell carcinoma of the prostate gland]. PMID- 11877978 TI - [Magnetic-resonance urography: experience os using small doses of Gd-DTPA and amplification of contrast techniques]. PMID- 11877980 TI - [Immunological aspects of acute pyelonephritis]. PMID- 11877981 TI - The age pattern of mortality in the 1918-19 influenza pandemic: an attempted explanation based on data for England and Wales. PMID- 11877982 TI - Edwin Chadwick and the poverty of statistics. PMID- 11877983 TI - Diphtheria and Australian public health: bacteriology and its complex applications, c. 1890-1930. PMID- 11877984 TI - The Derbyshire General Infirmary and the Derby philosophers: the application of industrial architecture and technology to medical institutions in early nineteenth-century England. PMID- 11877985 TI - Smith v. Clark and Clark v. Smith: eighteenth-century Scottish doctors in dispute. PMID- 11877986 TI - Introductory lecture. Advanced laser spectroscopy in combustion chemistry: from elementary steps to practical devices. AB - In recent years a large number of linear and nonlinear laser-based diagnostic techniques for nonintrusive measurements of species concentrations, temperatures, and gas velocities in a wide pressure and temperature range with high temporal and spatial resolution have been developed and have become extremely valuable tools to study many aspects of combustion. Beside the nonintrusive diagnostics of technical combustion devices the kinetics and microscopic dynamics of elementary chemical combustion reactions can be investigated in great detail by laser spectroscopy. These investigations show, that a small number of relatively simple elementary steps like H + O2-->OH + O, H2O2-->2OH, O + N2-->NO + N, NH2 + NO- >H2O + N2, OH + N2H control a large variety of combustion phenomena and pollutant formation processes. Laminar flames are ideal objects to develop the application of laser spectroscopic methods for practical combustion systems and to test and improve the gas-phase reaction mechanism in combustion models. Nonintrusive laser point and field measurements are of basic importance in the validation and further development of turbulent combustion models. Nonlinear laser spectroscopic techniques using infrared-visible sum-frequency generation can now bridge the pressure and materials gap to provide kinetic data for catalytic combustion. Finally, the potential of laser techniques for active combustion control in municipal waste incinerators is illustrated. PMID- 11877987 TI - Infrared frequency-modulation probing of product formation in alkyl + O2 reactions. Part IV. Reactions of propyl and butyl radicals with O2. AB - The time-resolved production of HO2 in the Cl-initiated oxidation of iso- and n butane is measured using continuous-wave (CW) infrared frequency modulation spectroscopy between 298 and 693 K. The yield of HO2 is determined relative to the Cl2/CH3OH/O2 system. As in studies of smaller alkanes, the branching fraction to HO2 + alkene in butyl + O2 displays a dramatic rise with increasing temperature between about 550 and 700 K (the "transition region") which is accompanied by a qualitative change in the time behavior of the HO2 production. At low temperatures the HO2 is formed promptly; a second, slower production of HO2 is responsible for the bulk of the increased yield in the transition temperature region. In contrast to reactions of smaller alkyl radicals with O2, the total HO2 yield in the butyl radical reactions appears to remain significantly below 1 up to 700 K, implying a significant role for OH-producing channels. The slower HO2 production in butane oxidation displays an apparent activation energy similar to that measured for smaller alkyl + O2 reactions, suggesting that the energetics of the HO2 elimination transition state are similar for a broad range of R + O2 systems. A combination of QCISD(T) based characterizations of the propyl and butyl + O2 potential energy surfaces and master equation based characterization of the propyl + O2 kinetics provide the framework for explanation of the experimentally observed HO2 production in Cl initiated propane and butane oxidation. These calculations suggest that the HO2 elimination channel is similar in all reaction systems, and that hydroperoxyalkyl (QOOH) species produced by internal H-atom abstraction in RO2 can provide a path to OH formation. However, the QOOH formed by the energetically favorable 1,5 isomerization (via a six-membered ring transition state) generally experiences significant barriers (relative to the radical + O2 reactants) to the production of an oxetane + OH. In contrast, the barriers to forming OH + an oxirane or an oxolane, via 1,4 or 1,6 isomerizations, respectively, are generally below reactants. PMID- 11877988 TI - Rotational effects in broadening factors of fall-off curves of unimolecular dissociation reactions. AB - Strong collision fall-off curves of unimolecular dissociation and the reverse recombination reactions are calculated by using the statistical adiabatic channel/classical trajectory model (SACM/CT). This formalism properly accounts for angular momentum coupling of transitional modes with overall rotation. Calculations are made for linear molecules dissociating into linear fragments and atoms with randomly chosen properties of the transitional modes and for isotropic as well as anisotropic potentials. Analytical representations of center broadening factors as a function of molecular parameters are given. A comparison between fall-off curves from rigid activated complex RRKM theory, from the present loose activated complex SACM/CT model, and from CT calculations on an ab initio potential is made for the HO2-->H + O2 system. It is shown that, besides rotational effects, energy-dependent anharmonicities of the density of states also influence the shape of the fall-off curves in this system. PMID- 11877989 TI - Time-dependent master equation simulation of complex elementary reactions in combustion: application to the reaction of 1CH2 with C2H2 from 300-2000 K. AB - Computational simulations of the title reaction are presented, covering a temperature range from 300 to 2000 K. At lower temperatures we find that initial formation of the cyclopropene complex by addition of methylene to acetylene is irreversible, as is the stabilisation process via collisional energy transfer. Product branching between propargyl and the stable isomers is predicted at 300 K as a function of pressure for the first time. At intermediate temperatures (1200 K), complex temporal evolution involving multiple steady states begins to emerge. At high temperatures (2000 K) the timescale for subsequent unimolecular decay of thermalized intermediates begins to impinge on the timescale for reaction of methylene, such that the rate of formation of propargyl product does not admit a simple analysis in terms of a single time-independent rate constant until the methylene supply becomes depleted. Likewise, at the elevated temperatures the thermalized intermediates cannot be regarded as irreversible product channels. Our solution algorithm involves spectral propagation of a symmetrized version of the discretized master equation matrix, and is implemented in a high precision environment which makes hitherto unachievable low-temperature modelling a reality. PMID- 11877990 TI - Use of quantum methods for a consistent approach to combustion modelling: hydrocarbon bond dissociation energies. AB - An attempt has been made to use modern quantum methods to codify the data base concerning bond dissociation energies in hydrocarbons. Calculations have been performed using two hybrid DFT methods, the well-known B3LYP formalism and a newly developed alternative named KMLYP. CBS-Q has also been employed where possible. The combination of experimental uncertainty and theoretical limitations is less than completely satisfactory. However, within uncertainties that translate to a factor of two at 1500 K, many transferable quantities are elucidated. A hybrid method has been developed for the correction of DFT calculations using group additivity. Given that the philosophy behind this work is the understanding that all data bases must be optimised for specific applications, so that avoidance of large errors is more important than absolute precision, the results appear to be quite useful. We are particularly encouraged by the performance of the KMLYP method, given its ease of application to molecules of practical interest. PMID- 11877992 TI - A direct transition state theory based analysis of the branching in NH2 + NO. AB - A combination of high-level quantum-chemical simulations and sophisticated transition state theory analyses is employed in a study of the temperature dependence of the N2H + OH-->HNNOH recombination reaction. The implications for the branching between N2H + OH and N2 + H2O in the NH2 + NO reaction are also explored. The transition state partition function for the N2H + OH recombination reaction is evaluated with a direct implementation of variable reaction coordinate (VRC) transition state theory (TST). The orientation dependent interaction energies are directly determined at the CAS + 1 + 2/cc-pvdz level. Corrections for basis set limitations are obtained via calculations along the cis and trans minimum energy paths employing an approximately aug-pvtz basis set. The calculated rate constant for the N2H + OH-->HNNOH recombination is found to decrease significantly with increasing temperature, in agreement with the predictions of our earlier theoretical study. Conventional transition state theory analyses, employing new coupled cluster estimates for the vibrational frequencies and energies at the saddlepoints along the NH2 + NO reaction pathway, are coupled with the VRC-TST analyses for the N2H + OH channels to provide estimates for the branching in the NH2 + NO reaction. Modest variations in the exothermicity of the reaction (1-2 kcal mol-1), and in a few of the saddlepoint energies (2-4 kcal mol-1), yield TST based predictions for the branching fraction that are in satisfactory agreement with related experimental results. The unmodified results are in reasonable agreement for higher temperatures, but predict too low a branching ratio near room temperature, as well as too steep an initial rise. PMID- 11877991 TI - Low-energy paths for the unimolecular decomposition of CH3OH: a G2M/statistical theory study. AB - The potential energy surface (PES) of the CH3OH system has been characterized by ab initio molecular orbital theory calculations at the G2M level of theory. The mechanisms for the decomposition of CH3OH and the related bimolecular reactions, CH3 + OH and 1CH2 + H2O, have been elucidated. The rate constants for these processes have been calculated using variational RRKM theory and compared with available experimental data. The total decomposition rate constants of CH3OH at the high- and low-pressure limits can be represented by k infinity = 1.56 x 10(16) exp(-44,310/T) s-1 and kAr0 = 1.60 x 10(36) T-12.2 exp(-48,140/T) cm3 molecule-1 s-1, respectively, covering the temperature range 1000-3000 K, in reasonable agreement with the experimental values. Our results indicate that the product branching ratios are strongly pressure dependent, with the production of CH3 + OH and 1CH2 + H2O dominant under high (P > 10(3) Torr) and low (P < 1 atm) pressures, respectively. For the bimolecular reaction of CH3 and OH, the total rate constant and the yields of 1CH2 + H2O and H2 + HCOH at lower pressures (P < 5 Torr) could be reasonably accounted for by the theory. For the reaction of 1CH2 with H2O, both the yield of CH3 + OH and the total rate constant could also be satisfactorily predicted theoretically. The production of 3CH2 + H2O by the singlet to triplet surface crossing, predicted to occur at 4.3 kcal mol-1 above the H2C...OH2 van der Waals complex (which lies 82.7 kcal mol-1 above CH3OH), was neglected in our calculations. PMID- 11877993 TI - An experimental and theoretical study of the product distribution of the reaction CH2 (X 3B1) + NO. AB - Measurements of the product branching ratios of the reaction CH2 (X 3B1) + NO (1) are presented together with calculations of the thermal rate constant and branching ratios using unimolecular rate theory. The reaction was investigated experimentally at room temperature using FTIR spectroscopy. The yields of the main products HCNO and HCN were found to be gamma HCNO = 0.89 +/- 0.06, gamma HCN = 0.11 +/- 0.06. Other minor products could be rationalized by numerical simulations of the reaction system taking into account possible consecutive reactions. The potential energy surface for the reaction was characterized by quantum chemical calculations using ab initio and density functional methods. The proposed reaction pathways connecting reactants to products were explored by multi-channel unimolecular rate theory calculations to determine the CH2 (X) + NO capture rate constant and the rate constants for the different product channels as a function of temperature. The calculated capture rate constant of k = 2.3 x 10(13) cm3 mol-1 s-1 is in good agreement with experimental values at room temperature. Collisional stabilization of the initial H2CNO recombination complex was predicted to be negligible up to pressures of > 1 bar. For ambient pressures and temperatures up to 2000 K, HCNO + H were calculated as the dominating products, with gamma HCNO approximately 0.94 in agreement with the experiments. The channel to HCN + OH was calculated with 0.015 < or = gamma HCN < or = 0.05, only slightly below the experimental value. PMID- 11877994 TI - Reactions of methyl radicals with propene at temperatures between 750 and 1000 K. AB - The pyrolysis of propene, initiated by methyl radicals, has been studied in the temperature range 750-1000 K and at a pressure of 0.13 bar in a quasi-wall-free reactor using laser heating by fast vibrational-translational (V-T) energy transfer. This is a convenient method to study homogeneous high-temperature kinetics since the reactor walls remain cold. The radial temperature distribution in the reactor has been investigated by four different methods: a stationary heat balance, optical absorption, pressure rise, and the temperature dependence of the rate of an isomerization reaction. Methyl radicals were produced via the fast thermal dissociation of di-tert-butyl-peroxide and the products were analysed using GC-MS. The main products of the overall reaction of the model system propene and methyl (C3H6 + CH3) were isopentane (iso-C5H12) and but-1-ene (1 C4H8), whereas allene (C3H4), trans-but-2-ene (trans-2-C4H8) and cis-but-2-ene (cis-2-C4H8) were minor components, all showing a strong dependence on temperature. The product distribution and the temperature dependence were analysed by a kinetic model of 61 species and 166 reactions developed for the high-temperature oxidation of butane and the low-temperature oxidation of n pentane and isopentane. It was necessary to include a few missing reactions and to adjust some rate constants to make the modeling agree with the experimental investigations. This extended mechanism has to be evaluated further in forthcoming experiments. PMID- 11877995 TI - Crossed beam studies of elementary reactions of N and C atoms and CN radicals of importance in combustion. AB - The dynamics of some elementary reactions of N(2D), C(3P,1D) and CN(X2 sigma +) of importance in combustion have been investigated by using the crossed molecular beam scattering method with mass spectrometric detection. The novel capability of producing intense, continuous beams of the radical reagents by a radio-frequency discharge beam source was exploited. From angular and velocity distribution measurements obtained in the laboratory frame, primary reaction products have been identified and their angular and translational energy distributions in the center-of-mass system, as well as branching ratios, have been derived. The dominant N/H exchange channel has been examined in the reaction N(2D) + CH4, which is found to lead to H + CH2NH (methylenimine) and H + CH3N (methylnitrene); no H2 elimination is observed. In the reaction N(2D) + H2O the N/H exchange channel has been found to occur via two competing pathways leading to HNO + H and HON + H, while formation of NO + H2 is negligible. Formation of H + H2CCCH (propargyl) is the dominant pathway, at low collision energy (Ec), of the C(3P) + C2H4 reaction, while at high Ec formation of the less stable C3H3 isomers (cyclopropenyl and/or propyn-1-yl) also occurs; the H2 elimination channel is negligible. The H elimination channel has also been found to be the dominant pathway in the C(3P,1D) + CH3CCH reaction leading to C4H3 isomers and, again, no H2 elimination has been observed to occur. In contrast, both H and H2 elimination, leading in comparable ratio to C3H + H and C3(X1 sigma g+) + H2(X1 sigma g+), respectively, have been observed in the reaction C(3P) + C2H2(X1 sigma g+). The occurrence of the spin-forbidden molecular pathway in this reaction, never detected before, has been rationalized by invoking the occurrence of intersystem crossing between triplet and singlet manifolds of the C3H2 potential energy surfaces. The reaction CN(X2 sigma +) + C2H2 has been found to lead to internally excited HCCCN (cyanoacetylene) + H. For all the reactions the dynamics have been discussed in the light of recent theoretical calculations on the relevant potential energy surfaces. Previous, lower resolution studies on C and CN reactions carried out using pulsed beams are noted. Finally, throughout the paper the relevance of these results to combustion chemistry is considered. PMID- 11877996 TI - Energy transfer in combustion diagnostics: experiment and modeling. AB - Laser induced fluorescence (LIF) of OH (A 2 sigma +) is measured in several atmospheric-pressure flames using a short-pulse laser system (80 ps duration) in conjunction with an intensified streak camera. The two-dimensional signal detection technique allows one to simultaneously monitor rotational and vibrational relaxation as well as electronic quenching. Rotationally-resolved LIF spectra affected by energy transfer are compared with the results of a rate equation model and are found to be in reasonably good agreement. It is shown that a significant contribution of fluorescence detected by broad-band techniques is due to levels populated by vibrational energy transfer (VET). Implications for picosecond LIF techniques for the time-resolved, quench-free detection of OH are discussed. A detailed analysis is presented for fluorescence spectra originating from levels populated by VET after excitation of states in the OH (A 2 sigma +, v' = 2) level. PMID- 11877997 TI - Temperature fields during the development of autoignition in a rapid compression machine. AB - Temperature and concentration fields have been investigated in the cylindrical combustion chamber of a rapid compression machine (RCM) by schlieren photography, chemiluminescent imaging and planar laser induced fluorescence of acetone and of formaldehyde in a 2-dimensional sheet across the diameter. The timescale of particular interest was up to 10 ms after the piston has stopped. Experiments were performed in non-reactive and reactive conditions. Acetone was seeded in non reactive mixtures. Combustion was studied first in a system containing di-tert butyl peroxide vapour in the presence of oxygen. The decomposition of di-tert butyl peroxide generates methyl radicals, which are then oxidised if oxygen is present. The overall reaction is exothermic and is characteristic of a conventional thermal ignition. In addition, chemiluminescence, resulting from CH2O*, accompanies the oxidation process. The combustion of n-pentane was then investigated at compressed gas temperatures that spanned the range in which there is a negative temperature dependence of the overall reaction rate, typically 750 850 K. The response to thermal feedback in this more complex thermokinetic system can be the opposite of the "thermal runaway" that accompanies di-tert-butyl peroxide combustion. The purpose of making comparisons between these two types of systems was to show how the temperature field generated in the RCM is modified in different ways by the interaction with the chemistry and to discuss the implications of this for the spatial development of spontaneous ignition. As the piston of the RCM moves it shears gas off the walls of the chamber. This probably creates a roll-up vortex, but more importantly it also collects gas from the walls and moves it across the cylinder head pushing it forward into a plug at the centre. Thus, soon after the end of compression there is an adiabatically heated gas which extends virtually to the wall, but this incorporates a plug of colder gas at its core. Diffusive transport will occur, but the timescale is relatively slow, and the effect hardly shows until at least 10 ms post-compression. The consequence of "thermal runaway" on a timescale that is compatible with the development of this temperature field is that the reaction rate in the adiabatically compressed toroidal region accelerates faster than in the core, and goes to completion first. A somewhat similar pattern emerges during n-pentane combustion when the initial condition is set at the lower end of the negative temperature dependent range. By contrast, at adiabatically compressed gas temperatures close to the upper end of the negative temperature dependent region, the reaction rate in the cooler core develops faster than that in the surrounding zone, and the temperature difference is rapidly smoothed out. This does not lead to spatial homogeneity in all respects, however, because different rates and extents of reaction generate different concentrations of intermediates. This stratification has implications for the eventual spatial evolution of spontaneous ignition. PMID- 11877998 TI - NO reburning study based on species quantification obtained by coupling LIF and cavity ring-down spectroscopy. AB - NO reburning is studied in a low pressure (15 hPa) premixed flame of CH4-O2 seeded with 1.8% of NO. Measurements were carried out by using cavity ring-down spectroscopy (CRDS) and laser induced fluorescence (LIF) techniques. The temperature profile was obtained by OH-LIF thermometry in the A-X (0-0) band. The OH profile was determined by LIF and calibrated by single pass absorption. The NO concentration profile was obtained by LIF in the A-X (0-0) band and corrected for Boltzmann fraction and quantum yield variations. The absolute concentration profile was determined in the burned gases by CRDS allowing a direct experimental determination of the NO reburning amount. Finally CH and CN mole fraction profiles were obtained by CRDS by exciting rotational transitions in the B-X (0 0) bands of CH and CN around 387 nm. We found a peak mole fraction of 29 ppm for CH and 3.3 ppm for CN. This last result is in contrast with a previous study of W. Juchmann, H. Latzel, D. L. Shin, G. Peiter, T. Dreier, H. R. Volpp, J. Wolfrum, R. P. Lindstedt and K. M. Leung, XXVIIth Symposium (International) on Combustion, The Combustion Institute, Pittsburgh, 1998, p. 469, performed in a similar flame, which reported much lower levels of CN. In that study the absolute concentration of CN was indirectly obtained by LIF calibrated by Rayleigh scattering. In a second part, experimental species profiles are compared with predictions of the GRI 3.0 mechanism. Comparison between experimental and predicted profiles shows a good agreement particularly for CN and NO species. A qualitative analysis of NO reburning is then performed. PMID- 11877999 TI - Laser absorption spectroscopy diagnostics of nitrogen-containing radicals in low pressure hydrocarbon flames doped with nitrogen oxides. AB - Absolute concentration profiles of NH2 and HNO have been measured in low-pressure methane/air flat flames doped with small amounts of NO and N2O. Addition of a small amount of nitrogen oxides does not alter significantly the flame speeds, temperature profiles and other parameters of the relatively well-understood methane/air flames. Intracavity laser absorption spectroscopy (ICLAS) and cavity ring-down spectroscopy (CRDS) are high-sensitivity techniques used to measure absolute concentrations of minor species in flames. In this work ICLAS is used to monitor NH2 and HNO, whereas CRDS is used for temperature measurements using OH spectra in the UV range. The (090)-(000) and (080)-(000) bands of the A2A1-X2B1 electronic transition of NH2 and (100)-(000) and (011)-(000) bands of the A1A" X1A' transition of HNO are used. Methane flames of different equivalence ratios are used. NH2 and HNO are observed in the flame as well as in the zone surrounding the flame, closer to the walls of the low-pressure chamber where the burner is located. An absorption originating from the species in this zone can affect substantially the results of line-of-sight experiments. A slow flow of nitrogen through the optical window holders was added in order to separate the spectra of HNO originating from the central flame zone. Calculations based on the commonly used GRI-Mech chemical mechanism predict two maxima in the HNO concentration profile in the NO doped flames. The first is located in the vicinity of the burner, and the second is closer to the luminescence flame zone. We were able to observe the first maximum, and its measured location agrees well with prediction. On the other hand, GRI-Mech strongly underpredicts the observed absolute concentration of HNO in this maximum. The measured absolute concentrations of NH2 are in reasonable agreement with the GRI-Mech predictions. PMID- 11878000 TI - Experimental and modelling study of sulfur and nitrogen doped premixed methane flames at low pressure. AB - Laser-induced fluorescence (LIF) has been used to observe NS and NO in methane/oxygen/argon laminar flames at low pressure doped with ammonia and sulfur dioxide. NS profiles as a function of height above the burner have been measured for rich flames. The effect of adding various amounts of sulfur dioxide on the observed NO in the burnt gas region has been investigated for a variety of stoichiometries. The experimental measurements have been compared with PREMIX simulations using a detailed elementary reaction mechanism for nitrogen- and sulfur-containing species in a methane flame. Sensitivity analysis has been employed to highlight the important reactions for NS, NO and SO2. The results demonstrate significant uncertainties in currently best available rate data for important reactions involving sulfur-containing species. PMID- 11878001 TI - Detailed surface reaction mechanism in a three-way catalyst. AB - Monolithic three-way catalysts are applied to reduce the emission of combustion engines. The design of such a catalytic converter is a complex process involving the optimization of different physical and chemical parameters (in the simplest case, e.g., length, cell densities or metal coverage of the catalyst). Numerical simulation can be used as an effective tool for the investigation of the catalytic properties of a catalytic converter and for the prediction of the performance of the catalyst. To attain this goal, a two-dimensional flow-field description is coupled with a detailed surface reaction model (gas-phase reactions can be neglected in three-way catalysts). This surface reaction mechanism (with C3H6 taken as representative of unburnt hydrocarbons) was developed using sub-mechanisms recently developed for hydrogen, carbon monoxide and methane oxidation, literature values for C3H6 oxidation, and estimates for the remaining unknown reactions. Results of the simulation of a monolithic single channel are used to validate the surface reaction mechanism. The performance of the catalyst was simulated under lean, nearly stoichiometric and rich conditions. For these characteristic conditions, the oxidation of propene and carbon monoxide and the reduction of NO on a typical Pt/Rh coated three-way catalyst were simulated as a function of temperature. The numerically predicted conversion data are compared with experimentally measured data. The simulation further reveals the coupling between chemical reactions and transport processes within the monolithic channel. PMID- 11878002 TI - A study of the reaction of oxygen with graphite: model chemistry. AB - A considerable amount of research has been directed towards the mechanism of oxidation of graphite as a model reaction system and because of its industrial importance. A number of recent studies have been concerned with ab initio molecular orbital calculations on graphite including model chemistry and the reactions with molecular oxygen. This study is concerned with oxidation steps involving the attachment of molecular oxygen to the graphene, the formation of carbon monoxide and, in particular, the subsequent oxidation reactions. PMID- 11878003 TI - Small-angle X-ray studies of soot inception and growth. AB - The high spectral intensity of X-rays produced by the undulator at the Basic Energy Sciences Synchrotron Radiation Center of Argonne's Advanced Photon Source has allowed us to perform small-angle X-ray scattering (SAXS) studies of the initial distribution of soot particles formed by various fuels. SAXS provides an in situ probe of the morphology of soot in the region between 1 and 100 nm and complements the ex situ technique of electron microscopy. The basic aspects of SAXS and its potential are illustrated with measurement on a laminar flame of acetylene in air. The more complex fuel toluene has been studied in a flat-flame burner that supports a CH4/H2/air or CO/H2/air diffusion flame stabilized by N2 co-flow. This burner produces a nearly constant temperature region above the flame where the pyrolysis and combustion of the heavier fuels occurs. Kinetic information is obtained by performing measurements of the scattered intensity profile as a function of the height above the burner. These profiles have been reduced to give the mean radius and dispersion of a distribution of spherical particles. Mean radii between 0.8 and 18 nm have been observed. The smallest of these is a factor of ten smaller than previously detected with Lorentz-Mie scattering. Near 1550 K, the soot distribution found in toluene shows a distinct step behavior that is consistent with model calculations. PMID- 11878004 TI - Thermodynamic and kinetic issues in the formation and oxidation of aromatic species. AB - The chemistry of aromatic species is discussed in the context of detailed kinetic modelling of benzene and butadiene flames and stirred reactors featuring ethylene and mixed aromatic/ethylene/hydrogen fuels. The development of reliable detailed mechanisms depends on the accuracy of the underlying hydrocarbon chemistry and the present paper highlights some current issues in the formation and oxidation of aromatics. In particular, uncertainties pertaining to the rates and product distributions of a range of possible naphthalene and indene formation sequences are discussed from the basis of improved predictions of key intermediates. The naphthalene formation paths considered include initiation via C5H5 + C5H5, C6H5 + C4H4 and C7H7 + C3H3 reactions and results are assessed in the context of a number of tentative detailed and simplified sequences. It is shown that a number of possible formation channels are plausible and that their relative importance is strongly dependent upon oxidation conditions. Particular emphasis is placed on the investigation of formation paths leading to isomeric C9H8 structures. The latter are typically ignored despite measured concentrations similar to those of naphthalene. The rates of formation of C9H8 compounds are consistent with sequences initiated by C6H5 + C3H3 and C6H5 + C3H4 leading to indene through repeated isomerisation reactions. The current work also shows that reactions of the type C9H7 + CH3 and C9H7 + 3CH2 provide a mass growth source that link five and six member ring structures. PMID- 11878005 TI - The influence of fuel additives on the behaviour of gaseous alkali-metal compounds during pulverised coal combustion. AB - The alkali-metal vapour release during pulverised hard (bituminous) coal combustion was investigated in a semi-technical drop flow reactor in the temperature range 1100-1400 degrees C. Absolute concentrations of total gas-phase sodium and potassium species were determined using the in situ/on-line excimer laser induced fragmentation fluorescence technique (ELIF). Alkali-metal concentrations measured for the untreated coals were found to be in the range 0.1 to 4.7 ppm, depending on the temperature. As well as observing the temperature dependence, the effect of co-feeding defined amounts of silica and clay minerals was studied. In addition, to assist interpretation of ELIF measurements, ash samples were taken and analysed by scanning electron microscopy (SEM) and energy dispersive X-ray analysis (EDX). The additives lead to a pronounced binding of the alkali-metal species and suppression of the sharp temperature dependence observed without co-feeding. Therefore, the use of such getter materials can be confirmed as an effective way to remove corrosive alkali-metal species from the flue gas in pulverised coal combustion. PMID- 11878006 TI - A combined crossed molecular beam and ab initio investigation of C2 and C3 elementary reactions with unsaturated hydrocarbons--pathways to hydrogen deficient hydrocarbon radicals in combustion flames. AB - Crossed molecular beam experiments on dicarbon and tricarbon reactions with unsaturated hydrocarbons acetylene, methylacetylene, and ethylene were performed to investigate the dynamics of channels leading to hydrogen-deficient hydrocarbon radicals. In the light of the results of new ab initio calculations, the experimental data suggest that these reactions are governed by an initial addition of C2/C3 to the pi molecular orbitals forming highly unsaturated cyclic structures. These intermediates are connected via various transition states and are suggested to ring open to chain isomers which decompose predominantly by displacement of atomic hydrogen, forming C4H, C5H, HCCCCCH2, HCCCCCCH3, H2CCCCH and H2CCCCCH. The C2(1 sigma g+) + C2H4 reaction has no entrance barrier and the channel leading to the H2CCCCH product is strongly exothermic. This is in strong contrast with the C3(1 sigma g+) + C2H4 reaction as this is characterized by a 26.4 kJ mol-1 threshold to form a HCCCCCH2 isomer. Analogous to the behavior with ethylene, preliminary results on the reactions of C2 and C3 with C2H2 and CH3CCH showed the H-displacement channels of these systems to share many similarities such as the absence/presence of an entrance barrier and the reaction mechanism. The explicit identification of the C2/C3 vs. hydrogen displacement demonstrates that hydrogen-deficient hydrocarbon radicals can be formed easily in environments like those of combustion processes. Our work is a first step towards a systematic database of the intermediates and the reaction products which are involved in this important class of reactions. These findings should be included in future models of PAH and soot formation in combustion flames. PMID- 11878007 TI - Determination of the CH + O2 product channels. AB - The multichannel CH + O2 reaction was studied at room temperature, in a low pressure fast-flow reactor. CH radical was obtained from the reaction of CHBr3 with potassium atoms. The overall rate constant was determined from the decay of CH with distance, O2 being introduced in excess. The result, after corrections for axial and radial diffusion, is k = (3.6 +/- 0.5) x 10(-11) cm3 molecule-1 s 1. The OH(A2 sigma +) chemiluminescence was observed, confirming the existence of the OH + CO channel. The vibrational population distribution of OH(A2 sigma +) is 32% in the v' = 1 level and 68% in the v' = 0 level (+/- 5%). The relative atomic concentrations were determined by resonance fluorescence in the vacuum ultraviolet. A ratio of 1.4 +/- 0.2 was found between the H atom density (H atoms being produced from the H + CO2 channel and from the HCO dissociation) and the O atom density (O + HCO). Ab initio calculations of the transition structures have been performed, associated with statistical estimations. The estimated branching ratios are: O + HCO, 20%; O + H + CO, 30%; H + CO2, 30%; and CO + OH, 20%. PMID- 11878008 TI - A theoretical analysis of the reaction between propargyl and molecular oxygen. AB - The temperature- and pressure-dependent kinetics of the reaction between propargyl and molecular oxygen have been studied with a combination of electronic structure theory, transition state theory, and the time-dependent master equation. The stationary points on the potential energy surface were located with B3LYP density functional theory. Approximate QCISD(T,Full)/6-311++G(3df,2pd) energies were obtained at these stationary points. At low temperatures the reaction is dominated by addition to the CH2 side of the propargyl radical followed by stabilization. However, addition to the CH side, which is followed by one of various possible internal rearrangements, becomes the dominant process at higher temperatures. These internal rearrangements involve a splitting of the O2 bond via the formation of 3-, 4- or 5-membered rings, with the apparent products being CH2CO + HCO. Rearrangement via the 3-membered ring is found to dominate the kinetics. Rearrangement from the CH2 addition product, via a 4-membered ring, would yield H2CO + HCCO, but the barrier to this rearrangement is too high to be kinetically significant. Other possible products require H transfers and, as a result, appear to be kinetically irrelevant. Modest variations in the energetics of a few key stationary points (most notably the entrance barrier heights) yield kinetic results that are in good agreement with the experimental results of Slagle and Gutman (I. R. Slagle and D. Gutman, Proc. Combust. Inst., 1986, 21, 875) and of Atkinson and Hudgens (D. B. Atkinson and J. W. Hudgens, J. Phys. Chem. A, 1999, 103, 4242). PMID- 11878009 TI - [The fat and fatty acids content in selected sea fish]. AB - The fat content as well as fatty acids in baltic herring, mackerel and salmon from Norway has been determined. The fat content was 7%, 11.6% and 13.2% respectively. Both fat extracted from mackerel and salmon contained much more of polyunsaturated fatty acids in comparison to that extracted from herring. PUFA content in herring fat was 15% whereas mackerel and salmon 29% and 25% respectively. PUFA represented mostly of omega-3 family. PMID- 11878010 TI - [Sodium chloride in food rations and dinners in mass catering institutions]. AB - The sodium chloride content in meals given by mass catering institution in all over country in 1988-1998 years was estimated. This study included daily food rations from 183 mass catering institution as hospitals, sanatoriums for both children and adults, boarding schools, infant schools and social welfare homes. We assessed also school dinners from 422 randomized selected schools and dinners from 55 internal and 56 surgical departments of provincial and regional hospitals in Poland. The mass of each meal was evaluated and sodium chloride content by Mohr's method was assessed. In most cases the salt content by 100 g of meal of 1000 kcal was calculated. The dinners and daily food rations analyze showed that sodium chloride content in meals was much higher than value recommended by World Health Organization (WHO). Salt amount in daily food rations of both children and adults was above 16 g. This value didn't include salt added to meals by boarders. School dinners provided about 7-10 g of salt. The average sodium chloride content in hospital dinners was about 16-20 g. In each studied group the NaCl content per 100 g of meal was similarly high and was 0.7-0.9 g. The results of this study show that meals given by mass catering institutions can increase risk of hypertension, strokes and gastric cancers because of high sodium chloride content. PMID- 11878011 TI - Accumulation rate of polychlorinated biphenyls (PCBs) in dandelion (Taraxacum officinale) in the conditions of soil contamination with oil derivatives. AB - The present study concerns PCBs accumulation in soil at two depths as well as in Taraxacum officinale plants. An analysis of the distribution of different groups of PCBs congeners in plant and soil material revealed the highest content of hexa and hepta-CB fractions. Statistical test of pair comparison has revealed significant differences in accumulation rates for the following pairs: penta chlorinated congeners are accumulated at a higher rate than tetra-chlorinated ones and hexa-chlorinated congeners are accumulated at a higher rate than penta chlorinated ones. This phenomenon occurs in the area of railway junctions (areas heavily polluted with oil derivatives). PMID- 11878012 TI - [Usefulness of an immunoassay test TRAIT for detection of genetically modified Roundup ready soybean in food products]. AB - The test based on immunoassay TRAIT Test for the specific detection of Roundup Ready Soybean was used for reference material in the form of dried powdered soy beans contained 0, 0.3, 1.25, 2.5% of genetically modified material, for soy beans declared as Roundup Ready and for soy products from Warsaw market. The detection limit was approximately 0.1% GMO on dry weight basis. Experiment was also carried out on heated soybeans. The positive results was obtained since temperature was under 65 degrees C during 15 minutes of heating grounded beans; above this temperature specific protein was not recognisable by the antibody. The TRAIT Test should be regarded as a qualitative method and could be recommended for screening purposes. Investigation demonstrated that above mentioned test was useful for detection of protein of genetically modified soybean in unprocessed products. PMID- 11878013 TI - [The frequency of breakfast and snack intake by pupils of primary schools in Biala Podlaska]. AB - The aim of this work was an attempt to assess the frequencies of breakfast and snack intake by schoolchildren and to identity its determinants. The study was carried out on spring in 2000, and included 2758 boys and girls aged 10-12 years, schoolchildren from nine primary schools in Biala Podlaska. It was found that more than 11% of schoolchildren had no breakfast at home, moreover 43% of boys and 53% of girls had no snack at school. The correlation between snack consumption at school and subjectively perceived living standard of their parents was significant in girls only (r = 0.26; p < or = 0.05). PMID- 11878014 TI - [Investigation of the pharmacodynamic properties of the solution of boric acid]. AB - By virtue of experimental investigations that had been carried out on animals an essential biological action of the aqueous solution of boric acid on the organism could be stated. The solution containing 274.46 mg of H3BO3 in a litre, administered to rats for 30 days orally in an "ad libitum" dose, caused changes in the lipid metabolism and in the water-mineral balance of the organism and in the acid-base equilibrium of the blood, it did not affect the morphology and the parameters of carbohydrate and protein metabolism. No influence has been stated of the investigated boric acid, as used in guinea pigs, on the excretion of bile. Suitably diluted with a nutritive fluid the solution acted spastically on the peristalsis of the small intestine of the rabbit. PMID- 11878015 TI - [Effect of some ADH inhibitors on microsomal alcohol oxidizing system activity in vitro]. AB - The treatment of methanol intoxication usually focuses on prevention of methanol conversion to its toxic metabolites due to administration of ethanol or 4 methylpyrasole (4-MP). Nevertheless there is a need for new measures treatment of methanol intoxication. For this reason the influence of some alcohol dehydrogenase inhibitors on the activity of microsomal alcohol oxidising system (MAOS) with methanol as a substrate was assayed. In the present study MAOS activity was measured spectrophotometrically in vitro at physiological pH 7.4 and 37 degrees C, assaying the degree of methanol oxidation. The quantity of arising formaldehyde was measured according with the method of Nash. The source of enzyme was hepatic slices. Our results have shown that MAOS activity was inhibited to different extents by: 4-MP, isovaleric amide, cimetidine, DMSO, EDTA, o phenantroline, pyrasole and theophylline at concentrations of 0.01 mmol/l, 0.10 mmol/l 1.00 mmol/l. And methylene blue, acetyl-L-carnitine and penicillamine increase MAOS activity in the process of methanol oxidation. In summary, 4-MP which plays an important role as an antidote in methanol intoxication was not an effective MAOS inhibitor. EDTA was found to be a highly effective inhibitor at all investigated concentrations. PMID- 11878016 TI - Determination of carbon monoxide in blood by means of microdiffusion. AB - There was developed a little-work-consuming and non-expensive microdiffusion method of carboxyhaemoglobin determination in blood, in low range concentration. It can be used in the assessment of passive smokers' exposure to carbon monoxide. The amount of blood necessary for the determination was 0.15 ml. An average precision of the developed method of the carbon monoxide determination (converting into HbCO) for the concentration ranged from 1 to 5%, from 3 to 12% and from 5 to 20% was respectively 5.96, 4.09 and 2.14. PMID- 11878017 TI - [Spectrophotometric determination of iodide ions in reaction with chloramine T and bromate and bromide ions using amplification procedure]. AB - A method for the determination of iodide ions in reaction with chloramine T is presented. There has been developed a method for the quantitative determination of small content of iodide ions making use of amplification reaction with bromine reduced from bromate (V)--bromide system. PMID- 11878018 TI - How psychological science informs the teaching of reading. AB - This monograph discusses research, theory, and practice relevant to how children learn to read English. After an initial overview of writing systems, the discussion summarizes research from developmental psychology on children's language competency when they enter school and on the nature of early reading development. Subsequent sections review theories of learning to read, the characteristics of children who do not learn to read (i.e., who have developmental dyslexia), research from cognitive psychology and cognitive neuroscience on skilled reading, and connectionist models of learning to read. The implications of the research findings for learning to read and teaching reading are discussed. Next, the primary methods used to teach reading (phonics and whole language) are summarized. The final section reviews laboratory and classroom studies on teaching reading. From these different sources of evidence, two inescapable conclusions emerge: (a) Mastering the alphabetic principle (that written symbols are associated with phonemes) is essential to becoming proficient in the skill of reading, and (b) methods that teach this principle directly are more effective than those that do not (especially for children who are at risk in some way for having difficulty learning to read). Using whole-language activities to supplement phonics instruction does help make reading fun and meaningful for children, but ultimately, phonics instruction is critically important because it helps beginning readers understand the alphabetic principle and learn new words. Thus, elementary-school teachers who make the alphabetic principle explicit are most effective in helping their students become skilled, independent readers. PMID- 11878019 TI - Four decades of research on the Swedish large lakes Malaren, Hjalmaren, Vattern and Vanern: the significance of monitoring and remedial measures for a sustainable society. AB - The large lakes of Sweden, Malaren, Hjalmaren, Vattern and Vanern, have been subjected to water-quality monitoring for almost four decades. Physicochemical variables, plankton and benthic invertebrates have been regularly assessed. Hydrological and sediment conditions, macrophytes, fish, primary production, bacteria and attached algae have been periodically investigated. The human impact, including industrial activities, was reflected in excessive amounts of organic matter, nutrients, metals and persistent organic compounds. From the late 1960s all municipal sewage works in the catchments of the lakes were upgraded to the highest technical standard, including chemical precipitation of phosphorus, and phosphorus discharge from the sewage works was thereby reduced by 90-95%. In addition, industries were obliged to restrict discharge of harmful substances. The reactions of the lakes to the remedial measures are discussed as well as the value of various indicators. The studies were instrumental in designing a national lake monitoring program. Additionally, results from large lake monitoring have contributed to the establishment of national water-quality criteria including, physical, chemical, and biological indicators. PMID- 11878020 TI - Morphometry and hydrology of the four large lakes of Sweden. AB - The four largest Swedish lakes Vanern, Vattern, Malaren and Hjalmaren all have a tectonic origin with multidirectional fault fissures, which influence the topography of the bottoms and the creation of subbasins. All the four lakes have been subject to several glacials, the latest of which started to recede from central Sweden c. 10,000 years ago. The many differences between the four lakes can be explained by their shapes, sizes and the relation between the sizes of the drainage basins and the size of the lakes. This paper focuses on processes considered relevant to aquatic organisms of the lakes: water balance, water residence time, water temperature, water exchange between subbasins within the lakes, and other internal water movements. Links between pollution, water exchange and retention of nutrients are discussed with an example from Lake Vanern. PMID- 11878021 TI - Recovery from eutrophication: experiences of reduced phosphorus input to the four largest lakes of Sweden. AB - In-lake concentration changes of phosphorus (P) and nitrogen (N) in lakes Vattern, Vanern, Malaren, and Hjalmaren in response to diminished input has been examined from the mid-1960s onwards. In the former two deep and oligotrophic lakes with slow water renewal, drastic reductions in P-input from the middle of 1970s caused just minor reductions in P-concentration over a very long time. At the same time accumulation occurred in the water mass of inorganic N and possible reasons are discussed. In the latter two mesotrophic to hypertrophic lakes, two shallow basins in L. Hjalmaren showed slow recovery due to release of P from sediments. The same basins and two basins in L. Malaren have suffered from N deficiency, particularly during the pre-phosphorus reduction years, and nitrogen fixation was indicated. In two L. Malaren basins recovery of in-lake P concentrations was better than expected in comparison to the so-called IMSA-model for lake recovery from nutrient pollution. In the other five lakes/basins chlorophyll concentrations after 20 years were similar compared to those modeled. PMID- 11878022 TI - Phosphorus in tributaries to Lake Malaren, Sweden: analytical fractions, anthropogenic contribution and bioavailability. AB - Riverine phosphorus (P) concentration and P-transport to Lake Malaren, the third largest lake in Sweden, has been monitored for 35 years in 12 major tributaries. During a period of 15 months, complementary assessments of particulate P, suspended matter and dissolved reactive P were made. Particulate P comprised 64%, dissolved unreactive P 23% and dissolved reactive P 13% as flow-weighted means, with high seasonal variability. "Background" or "reference" P-transports in the streams were estimated by different methods and the anthropogenic contribution to P-transport was shown to be reduced over time. Potential algal availability of particulate P showed a mean availability of c. 45% for water draining arable and forested land, while the availability was higher for sewage discharge particulate P and algal P. Calculated total bioavailable P in tributaries was shown to equal reactive P measured on coarsely filtered water and it was indicated that the proportion of bioavailable P was higher during the period with high anthropogenic P-contribution than with a lower contribution. PMID- 11878023 TI - Sediment evidence of early eutrophication and heavy metal pollution of Lake Malaren, central Sweden. AB - Lake Malaren is the water supply and recreation area for more than 1 million people in central Sweden and subject to considerable environmental concern. To establish background data for assessments of contemporary levels of trophy and heavy metal pollution, sediment cores from the lake were analyzed. Diatom inferred lake-water phosphorus concentrations suggest that pre-20th century nutrient levels in Sodra Bjorkfjarden, a basin in the eastern part of Malaren, were higher (c. 10-20 micrograms TP L-1) than previously assumed (c. 6 micrograms TP L-1). Stable lead isotope and lead concentration analyses from 3 basins (S. Bjorkfjarden, Gisselfjarden and Askofjarden) show that the lake was polluted in the 19th century and earlier from extensive metal production and processing in the catchment, particularly in the Bergslagen region. The lake has experienced a substantial improvement of the lead pollution situation in the 20th century following closure of the mining and metal industry. The lead pollution from the old mining industry was large compared to late-20th century pollution from car emissions, burning of fossil fuels and modern industries. PMID- 11878024 TI - Macrophyte development and habitat characteristics in Sweden's large lakes. AB - The four largest Swedish lakes, Vanern, Vattern, Hjalmaren, and Malaren, were surveyed by means of aerial IR-color photography in the 1970s. Along small slope gradients in all lakes the emergent communities extended to a width of hundreds of meters, while the extent on steeper slopes was a few meters. Because of the fractionate shape, L. Malaren has a very long shoreline, which promotes vegetation growth in the transition zone between land and water. Sheltered conditions are numerous and water lilies and other floating leaved and freely floating plants are abundant. Emergent as well as submerged macrophytes grow vigorously in the northernmost part of the oligotrophic lake Vattern. In L. Vanern exposed shores pre-dominate, but sheltered bays and archipelagos are overgrown with reeds. In the shallow lake Hjalmaren monospecific stands of Phragmites australis are dominant. Recurrent surveys of L. Malaren and L. Vanern in the 1990s showed changes that could be connected to the watertable fluctuations, which have been stabilized since the first investigation. Reeds and water lilies expanded in sheltered parts of the lakes, while the distribution of reed stands growing at their deep limit decreased. PMID- 11878025 TI - The environmental consequences of alien species in the Swedish lakes Malaren, Hjalmaren, Vanern and Vattern. AB - Twenty alien species have become established in the lakes Malaren, Vanern, Vattern and Hjalmaren. Intentional introductions include fish and the signal crayfish from North America, ornamental plants, and the Canada goose. Unintentional introductions include the crayfish plague introduced with infected crayfish, the zebra mussel, and Chinese mitten crab introduced with ballast water. The introduction of pathogens and parasites, in particular the crayfish plague, to the lakes has had the greatest environmental and socioeconomic effects and has contributed to the decimation of the indigenous noble crayfish. The stocking of brown trout and salmon with origins from different biogeographical regions has contributed to the extinction of relict indigenous fish species in L. Vanern. Although major ecosystem damage caused by the introduction of alien species, with the exception of the crayfish plague, has not occurred in the four large Swedish lakes, local problems of considerable dignity occur occasionally. PMID- 11878026 TI - Human impact on the fish diversity in the four largest lakes of Sweden. AB - The four largest Swedish lakes, Vanern, Vattern, Malaren, Hjalmaren, host important commercial fisheries for char, salmon, trout, whitefish, vendace (cisco), perch, pike-perch, pike and eel, i.e. highly diverse biological resources. Case studies illustrate physical, chemical and biological impacts on some of these commercial species caused by constructions of dams and ship canals, eutrophication, and overexploitation. Although some original species have been lost and a few new species have been added, the recent human interference has basically caused major shifts in dominance of the fish community structures because of eutrophication, alterations in the abundance of eel or crayfish, and due to overfishing. The latter is in some cases caused by the Great Lake Fishery Paradox--in an environment with several predators and competitors, but with ample food resources, especially salmonid fish but also species like pike-perch may adapt a life history favoring growth over sexual maturation. If harvested at a conventional size these populations will decline rapidly due to too small spawning stocks. PMID- 11878027 TI - Phytoplankton and water quality characterization: experiences from the Swedish large lakes Malaren, Hjalmaren, Vattern and Vanern. AB - Phytoplankton and environmental variables have been monitored in the large Swedish lakes Malaren, Hjalmaren, Vattern and Vanern since the 1960s. Measures to reduce phosphorus input and industrial waste products were taken during the 1970s. The phosphorus loading was then reduced by 90-95% resulting in a halving of the phosphorus concentrations in the most affected basins. The phytoplankton community reacted rapidly with decreased biomasses of cyanobacteria in summer as well as decreased biomasses of spring diatoms and cryptophycean flagellates. Other reactions were a contracted period of water-bloom, an increased taxon richness, an increased evenness in the biomass over the growth season, and a change in the species size structure within the phytoplankton community. Furthermore, the species richness in the large lakes is compared in relation to lake characteristics. A presentation of the occurrence of toxic cyanobacteria in the lakes is also given. Maximum-minimum values of 13-0.1 micrograms microcystin L-1 are established in connection with water-blooms in Hjalmaren and Malaren. The use of phytoplankton as a monitoring variable to detect water-quality changes is outlined and assessment criteria are presented. PMID- 11878028 TI - Mercury in sediment and fish communities of Lake Vanern, Sweden: recovery from contamination. AB - The past effluents of mercury (Hg) into Lake Vanern were considerable. The consequences of, and recovery from these have been monitored through continuous measurements of mercury in sediment and fish. Mercury levels in lake sediments in the vicinity of the main source of mercury, a chloralkali plant on the northern shore, have only decreased by slightly more than a half since the mid-1970s, despite a radical decrease in effluents from the source, already during the 1960s. The mercury levels in pike (Esox lucius) have decreased to a similar extent during this time period. They are now about 30% higher in the worst affected parts of the lake compared to the least affected parts. Lower levels have been measured in perch (Perca fluviatilis) and salmonoid fish in the lake. Despite the increased presence of mercury in the sediment of Lake Vanern, the mercury levels in the fish of the lake are relatively low compared to fish in lakes situated in the same region, but not affected by any local mercury effluents. As calculated, the total fish biomass of L. Vanern holds less than 1000th of the amount of mercury contained in the upper, biologically active layers of the bottom sediment of the lake. This demonstrates the potential influence of various environmental factors and motivates continued monitoring of mercury levels in the lake in the future. PMID- 11878029 TI - Monitoring of persistent organic pollutants (POPs): examples from Lake Vattern, Sweden. AB - Lake Vattern in southern Sweden is a large oligotrophic lake with high surface to catchment area ratio (ca. 0.4) and a water residence time of 60 years. The lake combines sensitivity to atmospheric POP-pollution with general susceptibility for effects of POPs and slow concentration decline. Time series, from the 1960s until 1996, of PCBs and DDT in fish and data on TCDD in sediment of L. Vattern are presented and compared to other large lakes. The long time dataset of POPs in Arctic char (Salvelinus salvelinus) shows a significant annual decline of about 5% for PCBs and 13% for DDT. These rates are in agreement with other studies. Nutrients as well as biological effects and factors affecting the fate of POPs in oligotrophic lakes with long residence times are discussed. We propose that oligotrophic clear lakes are important ecosystems for monitoring POPs in biota. PMID- 11878030 TI - Factors affecting population fluctuations of the glacial relict amphipod Monoporeia affinis (Lindstrom) in Sweden's largest lakes. AB - Factors affecting long-term (1982-2000) population densities of the glacial relict amphipod Monoporeia affinis were studied in Sweden's three largest lakes. Monoporeia showed large population fluctuations in all three lakes, with conspicuous peaks in density occurring in Lakes Vanern and Malaren. In Lake Vanern, amphipod densities showed highly significant relationships with spring maximum diatom biovolume at a 1-yr lag. The lack of relationship between diatom biovolumes and Monoporeia densities in L. Vattern is likely due to the larger depth and the lower nutrient content of this lake. In eutrophic L. Malaren, summer hypoxia (< 4 mg O2 L-1) is likely an important regulating factor. Hypolimnetic temperature showed a clear periodicity with relatively warm deep water occurring between 1989 and 1994. Hypolimnetic temperatures in Vanern and Vattern were correlated with total solar irradiance. However, neither hypolimnetic water temperature nor diatom biovolumes correlated with the North Atlantic Oscillation winter index. We speculate that variations in temperature and near-bottom oxygen concentrations negatively affect population densities by acting on recruitment success (reproduction) and juvenile (young-of-the-year) survival. PMID- 11878031 TI - Recruitment of pelagic fish in an unstable climate: studies in Sweden's four largest lakes. AB - Pelagic fish population biology was studied in the large Swedish lakes Vanern, Vattern, Malaren and Hjalmaren. It is crucial for fish fry in temperate regions to hatch early in the growth season to survive, and achieve large size before winter, and it is suggested that the key factors are to match the spring development of phyto- and zooplankton, but to avoid predation. This is more easily accomplished by the studied spring spawners smelt (Osmerus eperlanus) and pike-perch (Stizostedion lucioperca) than autumn spawners, such as vendace (Coregonus albula). It is shown that hatching of vendace fry shortly after ice break-up is beneficial for year-class strength. In oligotrophic large lakes with few predatory species a rapid increase in water temperature after ice-break is also promoting recruitment, whereas this is not the case in eutrophic lakes where predation pressure from other species may become too high. The results indicate that autumn spawners will have difficulties in adapting to global warming and it is also suggested that the life history can explain the large variations observed in year-class strength between years. PMID- 11878032 TI - Warmer winters: are planktonic algal populations in Sweden's largest lakes affected? AB - Winters in Sweden have become warmer in the 1990s, and as a consequence the timing of ice break-up and the growth and decline of spring phytoplankton has shifted, starting earlier. Even spring temperatures have become warmer, leading to an earlier beginning of the summer phytoplankton growth. The spring-ward shift in phytoplankton population growth has resulted in an extension of the growing season by at least one month. Although mean total phytoplankton biomass from May to October has not increased, the spring and early summer biomass of temperature sensitive phytoplankton groups, such as cyanobacteria and chlorophytes, has increased in the 1990s. No increase was noted for other phytoplankton groups. Considering that some species of cyanobacteria that commonly occur during a summer bloom, such as Anabaena, Aphanizomenon, and Microcystis, can be toxic, the effect of warmer winters on aquatic ecosystems is potentially far-reaching. PMID- 11878033 TI - Evidence-based practice in cancer nursing. Introduction. PMID- 11878034 TI - Strategies to translate research into practice. AB - OBJECTIVES: To describe potential strategies that may be useful in translating research into practice and to review the findings of recent reviews of the research conducted within medicine. DATA SOURCES: Review articles, textbooks, research studies, and guidelines. CONCLUSIONS: The most effective way to change physician behavior is through multifaceted strategies tailored to the particular setting. Strategies with consistent effect are education outreach visits and reminders. IMPLICATIONS FOR NURSING PRACTICE: The generalizability of these findings to nursing is not known. However, appraising this body of knowledge in medicine should help nursing's future efforts to investigate how to best translate research into practice. PMID- 11878035 TI - Increasing the visibility of patient fatigue. AB - OBJECTIVES: To describe the process used by our cancer center to assess staff knowledge and attitudes about cancer-related fatigue, and to change clinical practice of fatigue assessment and management. DATA SOURCES: Review articles, research studies, guidelines, and questionnaires. CONCLUSIONS: Change in clinical practice occurs slowly. One major change that occurred was the inclusion of a fatigue question in the Daily Patient Care Record. This inclusion signified a commitment by the institution to address this under-reported and under-treated symptom. IMPLICATIONS FOR NURSING PRACTICE: Professionals challenged to pursue practice change must be passionate, persistent, and patient. A multidisciplinary effort is needed to support change. PMID- 11878036 TI - Management of oral complications. AB - OBJECTIVES: To explore what nurses were doing to prevent or treat oral complications related to cancer and cancer therapies. DATA SOURCES: Review articles, research studies, consensus statements, and data collection. CONCLUSIONS: Different practice patterns exist between physicians and nurses in providing oral care interventions. Nurses initiated less toxic and less costly interventions than those prescribed by physicians. Standard assessment parameters are used infrequently. IMPLICATIONS FOR NURSING PRACTICE: Oral mucositis can be distressing by interfering with comfort, appetite, communication, and general well-being. Nurses need to initiate appropriate therapy for patient care. PMID- 11878037 TI - Processes and outcomes of evidence-based practice. AB - OBJECTIVES: To describe knowledge utilization, the models created to apply knowledge and promote evidence-based practice, and outcomes of integrating evidence into practice. DATA SOURCES: Textbooks, research and review articles, and professional experience. CONCLUSIONS: Evidence-based practice is both a process, which requires activities to evaluate evidence, and a product, which is the translation of evidence into a practice change. IMPLICATIONS FOR NURSING PRACTICE: The integration of evidence into nursing practice will strengthen nursing's theoretical base, decrease the variation in processes of care, improve patient outcomes, empower nurses, and help identify areas for research. PMID- 11878038 TI - Quality improvement of painful peripheral neuropathy. AB - OBJECTIVES: To use quality improvement methodology to improve the assessment and treatment of neuropathic pain caused by chemotherapy-related nerve injury. DATA SOURCES: Review and research articles, assessment scales, and textbooks. CONCLUSIONS: Assessment and treatment algorithms improved peripheral neuropathy related pain scores. Patient satisfaction with medication effectiveness decreased over time. Medication costs increased, but specialist-referral costs decreased. IMPLICATIONS FOR NURSING PRACTICE: Nurse-led quality improvement work can lead to positive changes in clinical practice. PMID- 11878039 TI - Dietary recommendations for neutropenic patients. AB - OBJECTIVES: To describe the development of an evidence-based practice project that will evaluate less restrictive dietary practices and focus on hand washing in adult patients with chemotherapy-induced neutropenia. DATA SOURCES: Guidelines, protocols, and published articles. CONCLUSIONS: There is a lack of scientific basis for food restrictions, a wide variation in policies related to low-microbial diets, and inconsistent compliance with restricted diets. Furthermore, lack of consistent practice has not been related to incidence of infection. IMPLICATIONS FOR NURSING PRACTICE: Evaluation of an evidence-based project such as this can lead to change in practice and institutional policy. PMID- 11878040 TI - Guidelines for the recurrence of pediatric cancer. AB - OBJECTIVES: To provide practice guidelines to assist patients and parents in coping with a return of disease. DATA SOURCES: Delphi studies, review, and research articles. CONCLUSIONS: When a child's cancer recurs, the patient and parents are at risk of physical and psychologic difficulties. Guidelines to assist patients and parents to cope have shown positive response. IMPLICATIONS FOR NURSING PRACTICE: Practice guidelines to assist patients and parents cope with a return of cancer can help health care professionals develop interventions for particular families. PMID- 11878041 TI - Cancer education: using the evidence. AB - OBJECTIVES: To discuss how a process improvement team analyzed the critical issues of education and support of patients and implemented change within a three hospital system. DATA SOURCES: Research and review articles, textbooks, guidelines, and standards. CONCLUSIONS: Education of cancer patients within three acute care hospitals was found to be highly variable and often inadequate. Our experience to improve the patient education process and coordinate print materials led to coordinated and effective evidence-based practice. IMPLICATIONS FOR NURSING PRACTICE: Education about disease, treatment, and maintaining lifestyle are achievable outcomes with adequate education and support. Educational goals can be met through the use of evidence-based practice. PMID- 11878042 TI - Using evidence in central catheter care. AB - OBJECTIVES: To discuss a project that identified the causes of central venous catheter infections, implemented corrective interventions, and evaluated the effectiveness of interventions. DATA SOURCES: Research studies, review articles, data collection, and personal experience. CONCLUSIONS: An educational intervention and individual demonstration improved sterile technique during catheter care, decreased infection rates, and decreased length of hospital stay. IMPLICATIONS FOR NURSING PRACTICE: Education has a significant impact on patient care and outcomes. Nurses can provide optimal catheter care for lowering infection risks. PMID- 11878043 TI - Support for evidence-based practice. AB - OBJECTIVES: To discuss examples of the process for evidence-based practice (EBP) and suggest helpful mechanisms for its support. DATA SOURCES: Published review and research articles, textbooks, guidelines, and web sites. CONCLUSIONS: Evidence-based practice requires a step by step process. Obtaining resources for EBP is essential to identifying and implementing and/or sustaining clinical care based on the best evidence available. IMPLICATIONS FOR NURSING PRACTICE: Finding resources to conduct EBP activities is one of the more challenging aspects of this approach to maintaining and improving quality of clinical care. However, nurses have found creative ways to obtain needed resources to implement EBP. PMID- 11878044 TI - Future perspectives of evidence-based practice. PMID- 11878045 TI - New approaches to the management of Non-Hodgkin's lymphoma. PMID- 11878046 TI - Immunotherapy: a novel treatment for Non-Hodgkin's lymphoma. AB - OBJECTIVES: To provide an overview of the principles of immunology, including immunotherapy, and information about monoclonal antibody therapy in the treatment of non-Hodgkin's lymphoma. DATA SOURCES: Published literature. CONCLUSIONS: Immunotherapy is a form of disease treatment that enhances the immune system with the use of biologic agents. One such agent is the monoclonal antibody, which can be combined with a radioisotope (e.g., iodine-131 or yttrium-90) or chemotherapeutic drug to deliver treatment directly to tumor cells with less toxicity to normal cells. IMPLICATIONS FOR NURSING PRACTICE: As the field of monoclonal antibody therapy continues to grow, nurses will play an important role in treatment and patient education. Nurses need to become knowledgeable in this area of cancer treatment and gain a better understanding of basic principles involved in immunotherapy. PMID- 11878047 TI - Monoclonal antibodies conjugated with radioisotopes for the treatment of Non Hodgkin's lymphoma. AB - OBJECTIVES: To describe the use of monoclonal antibodies labeled with iodine-131 tositumomab and yttrium-90 ibritumomab-tiuxetan as treatment for non-Hodgkin's lymphoma. DATA SOURCES: Literature review of trials of radioimmunotherapy. CONCLUSIONS: Radiolabeled monoclonal antibodies recognize and react with specific antigens to provide targeted therapy to tumor cells, particularly hematopoietic tumor cells. This therapy delivers greater amounts of radiation to malignant cells than normal cells and spares critical organs that do not express the antigen. Strict safety and patient precautions must be in place with the use of radiotherapy. IMPLICATIONS FOR NURSING PRACTICE: As the use of radiommunotherapy in the treatment of non-Hodgkin's lymphoma grows, the role of nursing in the care of these patients will also grow. Nurses will play an important part in patient and family education, as well as staff preparation. PMID- 11878048 TI - Establishing a radioimmunotherapy outpatient care clinic for Non-Hodgkin's lymphoma. AB - OBJECTIVES: To describe an outpatient treatment model for treating non-Hodgkin's lymphoma with radioimmunotherapy. DATA SOURCES: Experiences of The Hoag Cancer Canter (Newport Beach, CA) in developing an outpatient treatment model. CONCLUSIONS: The Hoag Cancer Center has put an outpatient radioimmunotherapy treatment model into place. Goals include providing coordinated, collaborative care, safe delivery of radioimmunotherapy in an outpatient setting, and establishing an education program for the medical team, patient, and patient's family. IMPLICATIONS FOR NURSING PRACTICE: In providing outpatient treatment for non-Hodgkin's lymphoma, nurses will have many functions, including educating patients and family, coordinating treatment schedules, adhering to radiation safety guidelines, and reviewing post-therapy and long-term side effects that might occur at home. PMID- 11878050 TI - [Characteristics of patients with arterial hypertension in Moscow (as exemplified by a municipal cardiological center)]. AB - AIM: To characterize hypertensive patients, residents of Moscow, treated at Moscow Municipal Cardiological Center (MMCC) in 1999. MATERIAL AND METHODS: A total of 1056 case histories of patients with the diagnosis of arterial hypertension or essential hypertension, directed to MMCC in 1999, were analyzed. The files were processed using specially developed standard methods. RESULTS: Patients with stage III essential hypertension aged over 60 years are most often treated at MMCC. Patients with more severe left-ventricular hypertrophy more often have cardiovascular complications. CONCLUSION: High incidence of cardiovascular complications indicates that the treatment of patients in an outpatient setting is poor. PMID- 11878049 TI - Current therapeutic approaches in the treatment of Non-Hodgkin's lymphoma. AB - OBJECTIVES: To review the causes, classification, treatment, and new treatment modalities of non-Hodgkin's lymphoma. DATA SOURCES: Published literature and experimental therapies. CONCLUSIONS: The increasing incidence of non-Hodgkin's lymphoma can be attributed to a growth in the number of immunodeficiency and autoimmune disorders, infectious agents, and human T-cell leukemia-lymphoma virus. Treatment options include watch-and-wait, radiation, chemotherapy, biologic therapy, and stem cell/bone marrow transplant. New therapies include the use of monoclonal antibodies and radioimmunotherapy. Experimental therapies include high-dose radioimmunotherapy and stem cell transplantation, vaccines, and antisense antiangiogensis agents. IMPLICATIONS FOR NURSING PRACTICE: Nurses will need to become versed in this modality, and will play an important part not only in therapy but also in patient education. PMID- 11878051 TI - [Advantages of capoten chronotherapy of patients with hypertension in an outpatient setting]. AB - AIM: To compare the efficiency of a short course of preventive chronotherapy and traditional therapy with capoten in an outpatient setting. MATERIAL AND METHODS: Forty-two patients with stage II essential hypertension were divided into 2 groups: 20 controls were treated with capoten in a dose of 12.5-50 mg 3 times a day and 22 received capoten once a day in the same single dose 1.5-2 h before arterial pressure (AP) acrophase. Central hemodynamics was studied by echocardiography, regional hemodynamics by reheoencephalography before and at the end of therapy. RESULTS: In the control group AP normalized after capoten therapy in 3 patients, decreased by at least 10% but did not normalize in 9, and did not change in 8 patients. The results in the chronotherapy group were as follows: AP normalized in 5, decreased by at least 15% in 13, and did not change in 4 patients. Normalization of daily profiles of hemodynamic values and more favorable shifts in the cerebral bloodflow were more often seen in patients who received chronotherapy. CONCLUSION: Capoten chronotherapy of patients with essential hypertension, carried out in an outpatient setting, is obviously preferable to traditional treatment: no side effects were observed and a better hypotensive effect was attained with lower total dose. PMID- 11878052 TI - [Effectiveness of controlled basic therapy of patients with bronchial asthma]. PMID- 11878054 TI - [Articular syndrome in practice of a local therapist]. PMID- 11878053 TI - [Diagnosis and treatment of renal diseases in outpatient settings]. PMID- 11878055 TI - [Osteoporosis: diagnosis, current approaches to treatment, prophylaxis]. PMID- 11878056 TI - [Interrelations of hyperplasia of gastric mucosa and helicobacter infection (based on data from a polyclinic center]. AB - AIM: To examine a relationship of hyperplastic processes in the gastric mucosa (GM) to Helicobacter infections. MATERIAL AND METHODS: A hundred patients aged 44 to 86 years who had gastric polyps [Group 1, foveolar hyperplasia (n = 21); Group 2, hyperplasiogenic polyps (n = 65), Group 3, adenomas (n = 14)] underwent esophagogastroduodenoscopy with biopsy of the mucosa from the athrum, body, and fundus of the stomach to reveal Helicobacter pylori (Hp) and GM morphological changes. RESULTS: Hp-associated chronic gastritis was detected in 56% of the patients (66.7, 52.3, and 57.1% in Groups 1, 2, and 3, respectively). The rate of Hp seeding correlated with the magnitude of inflammatory and dystrophic changes. As the regulation of epithelial growth and differentiation impairs, the magnitude of GM structural alterations increase. CONCLUSION: The findings indicate that there may be a relationship between the frequency, degree of Hp infection and the pattern of GM morphological changes, which makes it expedient to use eradication therapy in this group of patients. PMID- 11878057 TI - [Hyperinsulinemia and insulin resistance in pathogenesis of atherosclerosis and ischemic heart disease]. AB - AIM: To confirm the pathogenetic relationship of hyperinsulinemia, insulin resistance, and coronary disease. MATERIAL AND METHODS: Thirty-nine coronary patients (male) were examined using lipid loading test, bicycle ergometry, coronarography, and measurements of insulin, cholesterol (CS), triglycerides (TG), very low density lipoproteins (VLDL), low density lipoproteins (LDL), high density lipoprotein (HDL) CS, apoA-1, and apoB. RESULTS: Blood levels of CS, TG, VLDL, LDL, HDL CS, apoA-1, apoB, and insulin were measured before and 3, 6, and 9 h after lipid loading in 39 coronary patients and 20 normal subjects. Coronarography showed initially high levels of insulin in coronary patients with pronounced changes. Insulin level drastically increased after insulin loading; increases in TG and apoB levels were the most pronounced, while the concentrations of HDL CS and apoA-1 decreased and did not normalize 6 h after lipid loading. CONCLUSION: The results confirmed the relationship between hyperinsulinemia, insulin resistance, and coronary disease. PMID- 11878058 TI - [Effect of phenofibrate treatment on endothelial dysfunction in patients with history of myocardial infarction in young age]. AB - AIM: To evaluate the severity of endothelial dysfunction in patients with a history of myocardial infarction in young age and its changes during therapy with lipantil 200 M. MATERIAL AND METHODS: The percentage of dilatation of the brachial artery during reactive hyperemia test, number of circulating desquamated endotheliocytes, and lipid metabolism parameters were evaluated in 40 men with a history of myocardial infarction before the age of 45 years and 40 healthy men. RESULTS: Endothelial dysfunction was detected in coronary patients: the percentage of dilatation of the brachial artery was decreased, the number of circulating endotheliocytes increased, and lipid metabolism disordered. After 3 month lipantil 200 M therapy the severity of endothelial dysfunction significantly decreased and lipid metabolism parameters improved. CONCLUSION: Patients with a history of myocardial infarction in young age develop endothelial dysfunction, whose severity decreases after hypolipidemic therapy. PMID- 11878059 TI - [Six-month xenical (orlistat) therapy of patients with stable angina pectoris concomitant with obesity and hyperlipidemia]. AB - AIM: To evaluate the efficiency of 6-month therapy with xenical (gastrointestinal lipase inhibitor) in combination with diet in patients with stable angina pectoris associated with obesity and hyperlipemia. MATERIAL AND METHODS: An open comparative randomized study of the efficiency of xenical in combination with diet was carried out in patients with stable angina pectoris concomitant with obesity and hyperlipemia. Thirty coronary patients aged 45-65 years with stable angina of effort (functional class I-II) with body weight index 28.1-45.6 kg/m2 (mean 33.5 kg/m2) were examined. All patients presented with dyslipemia (low density lipoprotein (LDL) cholesterol more than 4.14 mmol/liter, triglycerides (TG) more than 2.2 mmol/liter). Controls (n = 15) were treated with diets alone for 6 months. In the main group diets were supplemented by xenical in a dose of 360 mg/day. RESULTS: Body weight index decreased in both groups (by 9.9% in the main group and by 4.2% in the control). Body weight stabilization during 6 months of treatment and the fact that it was slow and gradual were essential. In patients treated with xenical total cholesterol level decreased by 10.9% and of LDL cholesterol by 12.2% after 6 months (p < 0.05). Changes in the levels of high density lipoprotein cholesterol and TG were insignificant. The drug did not affect the incidence of angina attacks and improved exercise tolerance after 6 month therapy. Blood biochemistry (transaminases, alkaline phosphatase, glucose, and creatinine) changed negligibly. No side effects were observed; all patients received a complete 6-month course. CONCLUSION: The results confirm that xenical (orlistat) can be used for long therapy of patients with stable angina of effort concomitant with obesity and hyperlipemia. PMID- 11878060 TI - [Outpatient therapy. Current tasks]. PMID- 11878061 TI - [Effect of long-term treatment with enalapril, losartan and their combination on the quality of life of patients with congestive heart failure]. AB - AIM: To study the effect a combination of enalapril and losartan on life quality in patients with congestive heart failure (HF). MATERIAL AND METHODS: One hundred and eighty six patients with NYHA functional classes II to IV HF were examined. The study inclusion criteria were as follows: a left ventricular (LV) end diastolic volume of > 160 ml, a LV ejection fraction of < 35%, sinus rhythm, a cardiothoracic index of > 0.55, no history data on prior treatment with an angiotensin-converting enzyme inhibitor (ACEI) and/or an AT1-antagonist, a patient's written free-will consent to participate in the study. The exclusion criteria were as follows: pacemaker migration, an artificial pacemaker, high degree block, atrial fibrillation, cerebral circulatory disorders. All the patients were divided into 4 groups and received basic therapy with cardicet, 60 120 mg/day, aspirin, 250 mg/day, furosemide, 80-440 mg/week, and digoxin, 0.25 0.5 mg/day. Group 1 comprised 60 patients who refused therapy with ACEI and/or AT1-antagonist despite that they had indications for their use and they had been convinced many times. In Group 2 (n = 82) enalapril, an ACEI, was added to the basic therapy. Its initial dose of 2.5 mg/day was given once and slowly incremented to the therapeutical one (10-20 mg/day). Group 3 patients (n = 56) on the basic therapy were additionally treated with the AT1-antagonist losartan in a daily dose of 25-50 mg. They were started on 12.5 mg a day. In group 4 the basic therapy was added by a combination of enalapril and losartan in the same doses. The follow-up was 48 weeks. The efficiency of the treatments was controlled by the personal questionnaires SF-36, Life with Heart Failure, by evaluating the magnitude of clinical HF manifestations and by estimating the total life quality inxed. The data were analyzed by assuming that all the patients received the treatments. RESULTS: As compared with the conventional therapy and the use of each drug alone, a combination of the ACEI enalapril and the AT1-antagonist losartan promotes a more significant increase in the satisfaction of the patients with their vital activity, in the critical rate of their self-assessment of the "internal picture" of disease, and leads to a greater improvement of the quality of their life as a whole. CONCLUSION: The ICAE-AT1-antagonist combination exerts a positive impact on life quality in patients with heart failure. PMID- 11878062 TI - [Efficacy of 12-week monotherapy with arifon-retard of patients with isolated systolic hypertension]. AB - AIM: To evaluate the effect of arifon-retard (Servie, France) in patients with isolated systolic arterial hypertension (ISAH). MATERIAL AND METHODS: The efficiency of 12-week monotherapy with indapamid (arifon-retard) in a daily dose of 1.5 mg on systolic, diastolic, and pulse arterial pressure (SAP, DAP, and PAP, respectively) was studied in 20 patients with ISAH. RESULTS: SAP decreased by 25.1 +/- 1.8, (p < 0.00009), DAP by 4.2 +/- 0.9 (p < 0.0008), and PAP by 20.9 +/- 2.1 mm Hg (p < 0.00009). Antihypertensive effect was observed in all patients, AP normalized in 30% after 4 weeks of therapy and in 70% after 12 weeks, DAP not decreasing below 70 mm Hg in any of the patients. Antihypertensive effect of indapamid on SAP and PAP additionally increased from week 4 to week 12. With sufficient antihypertensive effect on SAP, the drug did not induce an excessive decrease of DAP and did not affect heart rate and blood biochemistry. The drug was well tolerated, no side effects were observed. PMID- 11878063 TI - [Spectral analysis of cardiac rhythm as an indicator of the autonomic cardiovascular regulation]. AB - AIM: To study cardiovascular function at rest and during exercise by spectral analysis of heart rhythm in patients with spinal osteochondrosis and cardialgia, in patients with excessive body weight, and with cardial type neurocirculatory dystonia (NCD). MATERIAL AND METHODS: A total of 223 patients were examined. Spinal osteochondrosis with cardialgia was diagnosed in 80, excessive body weight in 78, and cardial type NCD in 65. Heart rhythm wave structure was evaluated using VKK-12 Ergometer complex before and after bicycle ergometry. RESULTS: Submaximal exercise did not induce changes in the heart rhythm wave structure of normal subjects. In cardial type NCD the intensity of first order slow waves increased during the recovery period, while in patients with spinal osteochondrosis and cardialgia and in those with excessive body weight, heart rhythm centralization index increased. PMID- 11878064 TI - [Prospects of use of the combined preparation logimax in patients with essential hypertension]. AB - AIM: To study the effectiveness and tolerance of a combined drug logimax in patients with essential hypertension (EH). MATERIAL AND METHODS: The antihypertensive activity and safety of logimax was evaluated in 15 EH patients at week 10 of therapy. RESULTS: Logimax has a good antihypertensive effect for 24 hours in 86.7% patients. This was evidenced by the lowered mean systolic and diastolic pressures shown during 24-hour monitoring. Logimax was well tolerated. There were no side effects in 73.3% of the patients. CONCLUSION: The combined drug logimax is indicated for patients with EH as an antihypertensive agent to treat and prevent cardiovascular events. PMID- 11878066 TI - [beta-Adrenoblockers: current approaches to use]. PMID- 11878065 TI - [Parameters of bone tissue density in patients with bronchial asthma]. AB - AIM: To study bone density in patients with bronchial asthma (BA). MATERIALS AND METHODS: Bone densitometric indices were evaluated in 123 patients with BA in relation to the basic therapy, dosage, and the duration and severity of the disease. RESULTS: The bulk (59.3%) of BA patients is found to have the osteopenic syndrome whose detection rate increases with longer duration and greater severity. Decreased bone density is more evident with oral glucocorticoids (GCs) than with inhaled ones. The effect of topical GCs on bone density is determined by their bioavailability. The decrease in bone density is more noticeable in patients taking beclomethasone and minimal in those on basic fluticasone therapy. CONCLUSION: Considerable decreases in bone density in BA patients may be prevented by a rational basic therapy with first-line inhaled GCs with low bioavailability. PMID- 11878067 TI - [Theoretical problems of use of an immunomodulator likopid in clinical practice]. PMID- 11878068 TI - [Use of magnetic resonance tomography in angiology]. PMID- 11878069 TI - [Characteristics of anemia in rheumatoid arthritis]. PMID- 11878070 TI - [Training in a polyclinic setting as a factor promoting effective formation of physician thinking and personality of a physician]. PMID- 11878071 TI - [Quality of live in patients with hypertension]. PMID- 11878072 TI - [Contribution of Professor A N Kriukov to the development of therapeutic services and science in Uzbekistan]. PMID- 11878073 TI - Correlates of depression in older Latinos. AB - OBJECTIVE: To report the rates of depressive disorders--i.e., major and subthreshold depression--as well as the correlates of depression in a sample of older Latino primary care consumers. The study addresses the gap in the literature concerning depression and older Latinos residing in the U.S. DATA SOURCES AND STUDY SETTING: Data were collected from 150 Latino primary care consumers (50+ years-old) in Los Angeles County. Depression was measured using the depression module of the PRIME-MD Patient Health Questionnaire. Demographic, stress-related, health, and social integration data were also collected. STUDY DESIGN: A cross sectional design was employed vis-a-vis face-to-face interviews of respondents at the clinic sites or in their homes. Descriptive analyses and logistic regression modeling were used to describe the sample and to examine the correlates of depression. PRINCIPLE FINDINGS: Rates of depression indicate that 24.1% of the sample reported symptoms sufficient to meet the criteria for a PHQ depression diagnosis. Only social functioning and income were associated with the presence of a depressive disorder. Interference with social activities with family and friends as a result of physical and emotional problems was associated with a 1.86-fold increase risk of being depressed. CONCLUSIONS: Although most of the cases were classified as subthreshold, prior work has shown that subthreshold depression can be clinically significant and debilitating. Using brief screening instruments such as the PHQ, practitioners can identify cases needing further assessment and treatment. PMID- 11878074 TI - The parent they knew and the "new" parent: daughters' perceptions of dementia of the Alzheimer's Type. AB - Psychosocial death is a significant dimension of the Dementia of the Alzheimer's Type disease process but poorly studied. This article explores three phases of psychosocial death that emerged from in-depth interviews with three daughters caring for a parent with this chronic illness. The phases discussed include: (1) daughters creating a new relationship with their parent; (2) daughters grieving chronically throughout the illness; (3) daughters negotiating coherence between the parent that once was and the parent that exists now. Daughters' narratives reveal that witnessing the deterioration of a mind was a burdensome grave learning process that encompassed many losses. These daughters lacked appropriate education and support. Future research in this area is required to develop strategies for informal caregivers that respond to this disease process. PMID- 11878075 TI - Community-based case management: a model for outcome-based research for non institutionalized elderly. AB - Medical treatment and research has changed subsequent to a paradigm shift in fiscal management strategies for health care. The demand for greater fiscal and clinical accountability in health care has resulted in more bureaucratic control of physicians, health care providers and health care delivery. Institutional based care has been deferred to community-based care, and outcomes-based measurement of treatment interventions are becoming the benchmark of effective care. The increase in our elderly population's numbers and longevity of life, combined with fiscal and clinical constraints, invite a potential health care delivery crisis for our noninstitutionalized elderly. Interdisciplinary programs, such as community-based case management, that promote the health and well-being of our noninstitutionalized elderly can be an effective response to this crisis. However, the need for empirical evidence of their effectiveness is essential. PMID- 11878076 TI - Integrating behavior change theory into geriatric case management practice. AB - Case management practices have continued to grow despite a lack of clear evidence of their efficacy. With the expanding segment of the elderly population, there is a critical need to develop and identify programs that will address the many needs of the aging. Geriatric Case Management has been the avenue selected by many health care providers to address these issues, focusing on maintaining health status and improving linkages with medical and community resources. Studies testing the effectiveness of these models have failed to demonstrate their effectiveness in reducing depression, reducing acute care service use, and improving or maintaining health status. The Geriatric Case Management models presented in these lack an evidence-based, theoretical framework that provides definition and direction for case management practice. This article introduces behavior change theories as a method of structuring and delineating the case management intervention. The Transtheoretical Model and the Theory of Planned Behavior are discussed and methods of integrating these theories into practice are discussed. PMID- 11878077 TI - Physiatry, physical medicine, and rehabilitation: historical development and military roles. AB - The military has a rich tradition in relation to physical medicine and rehabilitation. The development of physical medicine and rehabilitation, to a large extent, can be traced to the wars of the 20th century. Military physiatrists were visionary leaders of our field, who responded to the overwhelming needs of war casualties, and with their insight, propelled psychiatry physical medicine and rehabilitation forward as a medical specialty. PMID- 11878078 TI - Rehabilitation of peripheral nerve injuries. AB - Traumatic injuries to peripheral nerves pose complex challenges to both military and civilian physicians. Treatment of nerve injuries must consider all aspects of the inherent disability. Pain control is of paramount importance. Little will be accomplished until pain is brought down to tolerable levels. Rehabilitation needs to be instituted as first-line treatment. Focus must be first placed on protection of the affected area from complications stemming from disuse and immobility and then on enhancement of strength, flexibility, sensory discrimination, and dexterity. Early intervention sets the stage for optimal physiologic and functional recovery. PMID- 11878079 TI - The rehabilitation of causalgia (complex regional pain syndrome-type II). AB - CRPS-type I or causalgia is a challenging pain syndrome and its pathogenesis remains controversial. Although its incidence is relatively low, the pain and suffering it causes can be severe and functionally debilitating. Early, accurate diagnosis permits initiation of appropriate therapeutic interventions and enhances the potential for successful treatment. PMID- 11878080 TI - Orthotics for the fighting force. AB - During peacetime, the military physiatrist uses orthotics in much the same way as the civilian physiatrist. In wartime, the focus shifts to a return-to-duty scope of care, and orthotic use is limited. This article discussed orthotic use in the wartime and military training/deployment setting with special emphasis on those orthoses that help soldiers successfully return to duty. Efficacy data are lacking on many commonly used orthotic devices, especially in the area of prevention and pain control. A good knowledge of the patient's functional and biomechanical limitations, use of a team-based approach to orthosis prescription, and consideration of efficacy studies and future trends will provide the best opportunity for a successful orthotic outcome. PMID- 11878081 TI - The military upper extremity amputee. AB - Throughout the course of military history, soldiers have continued to sustain amputation injuries during war times and during peacetime and training missions. What has changed over time is the etiology of, indication for, and management of the amputations. Technology has advanced significantly, often with some military connection. More work still needs to be done, especially in the areas of greater prosthetic limb function and usage as well as phantom pain and sensation management. Collaborative efforts among physiatrists, surgeons, prosthetists, and therapists can only benefit the patient. PMID- 11878082 TI - Landmine injuries and rehabilitation for landmine survivors. AB - There is a tremendous international need for the development of rehabilitation educational programs, especially in low-income countries. These programs are most urgently needed in those countries where there is active conflict, where there has been recent civil war, and where significant casualties have been created because of landmines and weapons-fire. Most of these countries are developing nations where rehabilitation is a lower priority than the provision of basic health needs. These countries generally have few financial, personnel, and system resources to provide much rehabilitation for their disabled citizens. Also, they generally have few, if any, physicians trained in the principles and practice of rehabilitation medicine. These countries can benefit from the use of psychiatric consultants to help assess their rehabilitation needs in designing relevant educational curricula for health professionals and in developing rehabilitation teams. These teams can then develop systems of care for the most common disabling conditions in that area of the world. Western technology, however, cannot be imposed on these countries. Any rehabilitation systems of care must be established within the context of the particular culture and governmental health system organization, if it is to be useful and succeed in a particular nation. PMID- 11878083 TI - Military lower extremity amputee rehabilitation. AB - The role of the military has changed over the past eight decades. Although the threat of global warfare still exists, the military of today faces new challenges as a peacekeeper in countries under turmoil. Despite these changes, the fear of bodily injury, such as limb loss and even the possibility of death, are real concerns for our active duty personnel. The military physician must be aware of the appropriate surgical, rehabilitative, and psychosocial needs of the lower extremity traumatic amputee. The physical medicine and rehabilitation specialist is a physician particularly suited to provide direction for and to oversee the overall care of the amputee's condition. An amputee center providing a multidisciplinary team approach has worked well for the military during war and peacetimes. The physical medicine and rehabilitation inpatient service at WRAMC continues the tradition and philosophy of the military amputee centers dating back to WWI. By growing with the fast-paced technologic advances in prosthetic devices, yet keeping true to the fundamentals of prosthetic prescription and design, physical medicine and rehabilitation in the military has remained at the forefront of amputee rehabilitation. Patient satisfaction with the rehabilitation process and his or her new artificial limb remain priorities whether the goal is to return to active duty or to resume an active lifestyle in the civilian world. PMID- 11878084 TI - Inpatient multitrauma rehabilitation in a U.S. military hospital. AB - Rehabilitation serves a vital role in the care of multiply injured patients in the military, from diagnosis of occult injuries to gaining functional independence. Optimal rehabilitative care of the war casualties is predicated on diligent and rigorous training and evaluation of similar care during peacetime. PMID- 11878085 TI - Rehabilitation of burn injuries. AB - Burn rehabilitation is a complex and difficult process for the burn survivor, his or her family, and the entire treatment team. This article describes the best approaches in burn care, including the use of a multidisciplinary team that emphasizes functional restoration through aggressive wound care, pain management, mobilization, and psychologic support. PMID- 11878087 TI - [Septic shock and selenium administration]. AB - Selenium is an essential trace element. In the form of selenocysteine, an amino acid, selenium is necessary for the activity of important enzymes (i.e. glutathione peroxidases, thioredoxin reductase). In the periodic table of the elements, selenium belongs to the same column as oxygen. In fact, seleno-enzymes have an important role in the detoxification of reactive oxygen species, especially peroxides and hydroperoxides. In septic and septic-like shock patients, reactive oxygen species, particularly peroxides, play an important role through their destructive actions, which are favourable as critical components of microbial destruction and also deleterious in excessive generation. This excessive generation results in tissue damage. Moreover, reactive oxygen species modulate the activation of important intracellular mediators (NF kappa B activation, arachidonic acid cascade). Simultaneously in patients with severe infection, there is a marked and early plasma selenium decrease. Redistribution due to selective selenium uptake for metabolic use could be one of the main mechanisms for this decrease. This review was carried out by questioning on the one hand the Medline database, by consulting the reviews and works available in the services of biology, biochemistry and pharmacy, by a prospective follow-up on the subject in Current Contents, but also thanks to library searches carried out by Aguettant laboratories. Several supplementary studies at various doses (from 140 to 1000 micrograms/day sodium selenite) have been conducted, though only on small groups of patients and with a questionable design. Selenium treatment seem to be promising in severely septic patients. However, in the absence of pertinent clinical data, only the administration of doses below adverse effect levels, staying within physiological limits, can presently be recommended (i.e. 200 to 500 micrograms/day of sodium selenite). PMID- 11878086 TI - [Clinical impact of atypical antipsychotics: prospective 6-month study of inpatients treated with risperidone or olanzapine]. AB - A survey of clinical and therapeutic evolution in patients treated with an atypical antipsychotic, risperidone or olanzapine, was carried out at La Colombiere Psychiatric Hospital, Montpellier. This prospective observational study was conducted over 6 months. Forty-nine patients were included in the olanzapine treatment group and thirty-five in the risperidone treatment group. The Clinical Global Impression (CGI) scale was used for evaluation at day 0, 7, 30, 90 and 180 of treatment. The majority of patients were improved after 30 days of treatment, and clinical improvement was observed in about 60 per cent of patients among those treated at day 180. The main therapeutic benefit occurred in respect of positive psychotic symptoms. Patients were discharged after 30 days of treatment. The atypical antipsychotic was not used with an additional neuroleptic for all patients. Tolerance was good, particularly for olanzapine. On the other hand, according to our results, others studies have to be performed to confirm the impact of the treatment on weight gain, assessment of subjective elements such as sedation, and the direct effect of the atypical antipsychotics on primary negative symptoms. PMID- 11878088 TI - [Pharmacokinetic studies in neonatology: regulatory and methodologic problems]. AB - Pharmacokinetic studies in neonates are essential because of maturational changes in physiological functions during this period. International harmonization conferences have established guidelines for such studies, including recommendations on age ranges to be studied and description of methods and study design. Multiple blood sampling to obtain a full pharmacokinetic profile cannot be proposed in neonates. Population pharmacokinetic analysis, with minimal sampling schemes, is more appropriate but involves more subjects than the traditional approach. Population pharmacokinetic analysis also focuses on the influence of individual factors on pharmacokinetic parameters. Most studies performed with neonates have involved drugs with a narrow safety margin and cleared by renal or metabolic routes to study the impact of immaturity on drug concentrations and on clinical effects. PMID- 11878090 TI - [Methodological reflections on 20 randomized clinical hydrotherapy trials in rheumatology]. AB - The aim of this study was to estimate the level of scientific evidence contributed by randomized clinical trials in rheumatologic indication of spa therapy. A literature search was made of computer data banks, with analysis of 20 randomized hydrotherapy trials. On the 20 identified randomized hydrotherapy trials, four were carried out double-blind; nine included a comparison of inter groups evolution. The indications assessed are chronic low back pain, osteoarthritis of the knee and hip joints, osteoarthritis of fingers, fibromyalgia, rheumatoid arthritis and psoriasis arthritis. The results suggest durable, persisting improvement several months after balneological care, in accordance with the following evaluation criteria: pain, handicap, quality of life, consumption of analgesics and of nonsteroidal anti-inflammatory drugs (NSAIDs). It was concluded that randomized evaluations, demonstrating a beneficial and prolonged clinical effect of balneologic treatments, exist in respect of the main indications for rheumatologic hydrotherapy. These results acknowledge the medical help given by hydrotherapy within the aforesaid parameters, to which may be added a reduction of the adverse gastrointestinal events of NSAIDs. Evaluation needs to be continued to clarify the medical benefit offered by rheumatologic hydrotherapy. PMID- 11878089 TI - [Pharmacokinetics after oral and intravenous administration of d,l-monolysine acetylsalicylate and an oral dose of acetylsalicylic acid in healthy volunteers]. AB - We studied the ASA pharmacokinetics of single doses of 500 mg and 1000 mg of D,L lysine-monoacetylsalicylate (Lys-ASA) administered both orally (Delgesic) and 500 mg parenterally (Aspisol) as well as 500 mg acetylsalicylate (ASA, Aspirin) in 13 healthy volunteers. Blood samples were taken before and at defined times up to 48 h after application of Lys-ASA and ASA. Analysis for ASA and its metabolite salicylic acid were performed by HPLC. All concentration versus time data were presented descriptively. As far as ASA was concerned, differences were assessed by means of ANOVA according to Friedman including post hoc Wilcoxon tests for each time point. Pharmacokinetic parameters were calculated based on a one compartment model. The concentration vs. time curves after oral intake of 500 mg of ASA and Lys-ASA differed significantly (p < 0.001). Peak serum ASA concentrations (Cmax) were 6.8 mg/l for oral Lys-ASA and 2.7 mg/l for ASA per os. The corresponding tmax-values were 14.2 and 38.0 min. Absolute bioavailabilities for 500 mg doses were 75.4 and 63.4 pour cent, respectively. After intake of 100 mg and 1000 mg oral doses of Lys-ASA Cmax was 2.7 mg/l and 15.9 mg/l, tmax being 14.2 min for the 1000 mg dose. The shortest half-life was found after i.v. injection with 7.5 min. Metabolism was fast with maximum rise of salicylic acid concentration after injection of Lys-ASS. We conclude that concerning time dimension oral administration of Lys-ASA is almost equivalent to i.v. Lys-ASA and may be an alternative for i.v. administration in cases of acute heart attacks. PMID- 11878091 TI - [Pain management: a complex therapeutic approach]. PMID- 11878092 TI - [The pharmacologic basis of pain treatment]. AB - The treatment of chronic pain uses drugs from different pharmacological classes. Analgesics are the common basis of these treatments. Peripheral analgesics (or minor analgesics such as paracetamol) and non-steroidal anti-inflammatory drugs are used for moderate pain (grade I of WHO). Major analgesics, opioids, are used for more severe pain (grades II and III). When pain can be related to a precise cause or location, more specific drugs may be used. This is done in migraine, facial pain, muscular spasms, dental pain, local inflammation. Chronic pain of grades II and III is treated with opioids. According to the severity, agents of different powers are used: partial agonists, full agonists of receptors OP3 (mu) and OP2 (kappa). According to other pathological signs linked to pain, coanalgesic drugs may be used in association: psychotropic drugs, either psycholeptic drugs which act synergistically with analgesics and bring their own effects, anxiolytic and/or neuroleptic, or anti-depressants which inhibit the depression state that may be associated with pain. Corticosteroids are also very useful for the numerous effects they induce: inhibition of the inflammation process, CNS stimulation, analgesics in medullary, or plexus compressions and in elevations of intracranial hypertension. Moreover their metabolic effects may be useful in cachectic states. The pharmacological treatment of chronic pain of grades II and III poses the problem of chronic administration of increasing doses of opioids and of their coprescription, of acquired tolerance, of dependence and of toxicity induced by drug accumulation. PMID- 11878093 TI - [Management strategy for the patient in chronic pain]. AB - Pain is the first consultation motive everywhere in the world. Modern medicine consider the fight against pain as a true challenge, and analgia has today become a real ethical imperative. The concept of chronic pain as a true disease, first described by Leriche, has given rise to Pain Centers. Pain Centers allow a multidisciplinary approach, meticulous evaluation with scales of pain and finally global management of the patient with chronic pain. PMID- 11878094 TI - [Pain in children: a pediatrician's point of view]. AB - The interest in the topic of pain in children is increasing within the paediatric community after a long period of misrecognition, often under-valuation, sometimes negligent or even denial. A lack of knowledge concerning pharmacology and toxicity of antalgic drugs and poorly adapted galenic presentations induced delay and complications in their use in children. The recent progress is supported by a better semiology, development of adequate pain scales, availability of adapted drugs, an effort at education of caregivers and public sensitization. In primary care the objective is to lead the practitioner to identify and consider pain in his/her preventive and curative strategies. In hospitals efforts are needed to improve the use of protocols and evaluations, the systematic practice of preventive analgesia and the quality of interprofessional cooperation within care teams. Managed care organizations are today an effective system to promote these practices. PMID- 11878095 TI - [Dispensation of opioids by pharmacists in Tunisia]. AB - This national survey aims to evaluate opioid availability and prescription by pharmacists in Tunisia and to examine pharmacists' attitude regarding Tunisian law. We surveyed a sample of 300 pharmacists randomly selected from the National Council of Pharmacists list and using the random table. This study started in September 1999 by sending to pharmacists a confidential questionnaire asking about the importance, the rate of sale and the availability of analgesics in their pharmacy. It also tried to determine pharmacists' opinions regarding Tunisian law. A total of 157 pharmacists out of 300 responded to the survey (52 per cent), 95 per cent were working in pharmacies and 15 per cent in hospitals. Analgesics were estimated to be important to very important in their work in 84 per cent and less important in 16 per cent of cases. They were given under advice with great importance in 85 per cent of cases and with less importance in 15 per cent of cases. Analgesic self-medication was frequent in 95 per cent of cases and rare in 4 per cent of cases. Analgesics of levels 1 and 2 were often to always available in 97 per cent of cases and rarely available in 1.5 per cent of cases. Some 84.7 per cent of pharmacists had opioid supplies and 8.9 per cent had no opioids in stock and 6.4 per cent hadn't given a response. 30 per cent of pharmacists think that the 7 days law for opioid prescription should be modified and 66 per cent think it should not, fearing illicit use, fraud and dependance. Pharmacists think that the minimal list of opioids to be stocked in a pharmacy is sufficient because of low demand. PMID- 11878096 TI - [Medical networks to better take care of inpatients and outpatients in pain. Experience of the French "pain network" of Basse Normandie]. AB - We describe here our own experience in creating a regional multidisciplinary network made up of health professionals involved in the treatment of pain. The rationale, aims, difficulty already experienced and still expected, and the results are discussed. PMID- 11878097 TI - [The saga of aspirin: centuries-old ancestors of an old lady who doesn't deserve to die]. AB - Where do analgics come from? If their ancestors are many centuries old, we observe that the four main drugs of modern analgesia, morphine (1816), codeine (1832), paracetamol (1893) and aspirin (1897) were discovered during the 19th century. And through what 'sagas'! The first known prescriptions, written on earthenware shelves in Mesopotamia 3 centuries BC, already mentioned medications derived from willow to cure headaches. The Greeks dedicated to Asclepios, god of therapeutics, a statue carved in a willow trunk as a symbol! Thus, before becoming a drug, aspirin was born from the willow, which grows with its feet in water 'without suffering', as the ancestors put it. But before it walked on the moon with Neil Armstrong in 1969, the discovery of aspirin as a drug was the consequence of the filial love of a young researcher, Felix Hoffmann, who wanted to decrease the resistant pain of his rheumatic old father. PMID- 11878098 TI - The management of adverse drug reactions: from diagnosis to signal. AB - Severe adverse reactions (ADR) are uncommon and benefits of drug treatment usually outweigh the disadvantages. Definitions and guidelines for managing adverse drug reactions (ADRs) are proposed mainly with the aim of helping to keep the possibility of ADR at the front of our minds and making the right diagnosis. As spontaneous reporting is a cornerstone in post-marketing surveillance, the reasons for under-reporting are thoroughly analysed. Finally, a reminder is given that communication of information should be encouraged within the medical community and that everyone who is involved in the treatment of patients has an active part to play in the network of pharmacovigilance. PMID- 11878099 TI - [Sudden death during anesthesia: human error, drug related or cardiac death?]. AB - We carried out a retrospective analysis of 1500 forensic autopsies following sudden unexpected cardiac death. This analysis showed a group of 43 cases that could have been related to surgery and/or anaesthesia. Pathological examination revealed the existence of cardiac lesions in 40 cases: arrhythmogenic right ventricular cardiomyopathy (14 cases), coronary artery disease (9 cases), cardiomyopathy (8 cases), structural abnormalities of the His bundle (7 cases), mitral valve prolapse (1 case) and acute myocarditis (1 case). These disorders are compatible with a paroxysmal (rhythmic) phenomenon at the origin of the cardiac arrest. Identification of the cause of death in a patient who was presumed to be at low risk may provide major relief to the patient's family and the medical staff. PMID- 11878101 TI - [Urinary lithiasis of medical origin]. AB - Analysis of 22,510 urinary calculi between January 1991 to July 2000 performed by infrared spectroscopy allows for separation of drug-induced urolithiasis into two categories: first, the drugs physically embedded in the stone (n = 238; 1.0 per cent), notably indinavir monohydrate (n = 126; 52.9 per cent), followed by triamterene (n = 43; 18.1 per cent), sulphonamides (n = 29; 12.2 per cent) and amorphous silica (n = 24; 10.1 per cent); second, the category of metabolic nephrolithiasis induced by drugs (n = 140; 0.6 per cent), involving mainly calcium and vitamin D supplementation (n = 56; 40.0 per cent) and carbonic anhydrase inhibitors (n = 33; 23.6 per cent). Composition of the stone depended not only on the inducer drug but also on the metabolic state of the patient. Today, drug-induced stones comprise about 1.6 per cent of all calculi in France. Physical analysis and therapeutic history recall of such patients are the keys to diagnosis. Medical care is based on drug avoidance or dose adjustment with increased diuresis and, if necessary, change in urinary pH. PMID- 11878100 TI - [Misuse and or overdose of BCG vaccine: evaluation over a 4-year period]. AB - The inappropriate use of BCG vaccine could be followed by adverse events (AE). In order to know their frequency and type, we evaluated the spontaneous reports of misuses (MS) and overdosages (OV) over a 4-year period. One hundred and twenty six notifications were collected; 109 concerned MS, 13 OV, and 4 concerned both MS and OV. We recorded a high number (97) of MS related to the injection of a BCG vaccine instead of a tuberculin test (TT). Among the 126 initial notifications, 122 cases of AE were reported, 107 after an MS, 11 after an OV and 4 after both MS and OV. The most common AE were related to injection-site reactions which were in fact expected reactions when BCG is used under correct conditions. In half of the cases systemic signs were associated. The most common were asthenia, fever and lymphadenopathy. Seven cases related to MS were considered as serious according to the ICH definition. Six of them recovered, for one the outcome was unknown. It appeared that perhaps the presentation of the products BCG vaccine and TT could play a role in contributing to this inappropriate use. Both presentations contain 10 doses but there are significant differences in colours used and in specific instructions. Despite the fact that the AE reported after an MS and/or an OV represented the one-third of the total notifications recorded with the BCG vaccine over the 4-year period under review, they have to be considered in light of the 170 million doses distributed in 150 countries. We have to consider the possibility of under reporting because of the delicacy of incorrect administration and differences in pharmacovigilance structures worldwide. Even if the number of AE is low, MS and OV should be reported and documented for good pharmacovigilance surveillance. PMID- 11878102 TI - [Therapeutic results of 5-fluorouracil in multiple and unresectable facial carcinoma secondary to xeroderma pigmentosum]. AB - A prospective study was done to test the efficacy of 5-fluorouracil (topical and systemic) in multiple and unresectable histologically proven facial squamous cell carcinomas (SCC) secondary to XP. Twelve patients (7M/5F, mean age 19.8 years) with multiple facial SCC were treated between 1994 and 1997. 5-FU was used as a twice-a-day local application in the documented areas, by continuous infusion associated with cisplatin (2 patients) and short infusion combined with folic acid (3 patients). Evaluation was done by clinical examination every two months for topical therapy and after every cycle for systemic treatment. Median treatment duration was 12 months (2 to 36 months). Treatment was well tolerated excluding episodes of pruritus in the treated areas. We observed mainly superficial tumour regression followed by dryness and crusting. In 5 cases, we performed biopsies after treatment showing in one case an extensive fibrosis with absence of tumour. However in the remaining 4 cases, despite a superficial reduction of tumour and a reconstitution of the epidermis, viable and unmodified squamous cell carcinoma remained in the deeper dermis. In the 5 patients treated by systemic 5-FU, we observed 1 complete response and 3 partial responses. Despite a dissociation between a good cosmetic result and a relatively superficial effect, topical 5-FU represents a useful therapeutic option in multiple unresectable facial SCC in patients with XP. Systemic chemotherapy is recommended in the event of more extended or profound lesions. PMID- 11878103 TI - [Breast cancer in the elderly woman: epidemiological data and series from the Centre Rene Huguenin]. PMID- 11878104 TI - [Hormone treatment for brest cancer in the elderly woman]. PMID- 11878105 TI - [The limitations of radiotherapy for breast cancer in the elderly patient]. PMID- 11878106 TI - [Surgery in the elderly person: specific risk--decision making criteria]. PMID- 11878107 TI - [Breast cancer imaging in the elderly female]. PMID- 11878108 TI - [Are there anatomo-pathological specificities in the elderly woman?]. PMID- 11878109 TI - [Relevance and indications of the sentinel lymph node in breast cancer in the elderly woman]. PMID- 11878111 TI - [Retrospective study of adverse drug reactions in the elderly: experience of regional center for pharmacovigilance]. AB - Cases of adverse drug reactions which occurred in the elderly, and were notified to the Regional Centre of Pharmacovigilance of Angers between 1995 and 1998, were analysed in two subgroups: from 75- to 84-years-old and older than 84 years. Among the 263 cases recorded in the elderly, there was no significant difference between the 75- to 84-year-old patients (180 cases) and the patients older than 84 years (83 cases) concerning past medical history, severity of effects or medication. Side-effects in patients over 74 were mainly cutaneous, haematological and neuropsychiatric, involving in decreasing order of frequency cardiovascular, neuropsychiatric, antibiotic, analgesic and anti-inflammatory drugs. In comparison with patients under 75, adverse drug reactions are more frequent and more serious, notably overdose and drug interactions, in elderly people. The occurrence of some avoidable side-effects justifies the strengthening of information about therapeutic safety in the elderly. PMID- 11878110 TI - [A new tool for evaluation of medication errors applied to pediatric hematology]. AB - This study was conducted in a haematological paediatric department and was aimed at evaluating drug medication errors. Their frequency was studied, but also and mainly their degree of severity and preventability. Only adverse drug events that were identified as possibly due to pharmacological properties of drugs or medication errors were collected. An original method was used, based on a multidimensional mathematical tool, called Factorial Analysis of Multiple Correspondences (FAMC), in order to assess the grade of severity and preventability for each adverse drug event. A total of 155 adverse drug events were detected for 34 out of 52 patients hospitalized during the study period. The prevalence rate was 65 per cent and among these adverse drug events, 16 per cent were serious and 53 per cent were avoidable. Apart from the fact that the FAMC helped to determine the grade of preventability, FAMC allowed one to demonstrate that allergy and medication errors were the most avoidable adverse drug events. In this way the method used was validated. This study permitted the assessment of drug medication errors in this department and helped to choose the priorities for the management of preventive actions. PMID- 11878112 TI - [Hepatotoxicity of a synthetic cortisol antagonist: OP'DDD (mitotane)]. AB - The adrenolytic agent, Op'DDD (Mitotane) has been employed for almost 50 years for treatment of Cushing's syndrome. Despite clinical observations of elevation of hepatic enzymes encountered in patients taking the drug, there are few published data regarding the frequency, time course and factors that might influence hepatic toxicity of Mitotane. We analysed 10 patients consecutively treated with Mitotane for Cushing's syndrome. We measured hepatic transaminase and gamma glutamyl transferase before, during and after treatment. The study population included 6 women and 4 males, with a mean age of 41 years. Seven patients presented Cushing's disease while two had adrenal tumours and one had an undetermined origin of Cushing's syndrome. After a progressive increase, patients were being treated with a mean dosage of 9 g per day. All patients had elevation of either GGT or ALAT and all but one had elevation of transaminase (the maximum increase was sixfold the basal value). The only variable correlated with hepatic increase was the body mass index. In contrast, the severity of the disease, alcohol intake, and other biological characteristics were not correlated with transaminase elevations. We conclude that transaminase increase is encountered in the vast majority of patients treated with Mitotane. Levels at which the drug should be withdrawn remain to be established. PMID- 11878113 TI - [Amenorrhea and galactorrhea after acute overdose with milnacipran]. PMID- 11878114 TI - [Acute intoxication with disopyramide: case report with literature review]. PMID- 11878115 TI - [Scientific rigor and clinical implications: two objectives reconciled]. PMID- 11878116 TI - [Monitoring expired oxygen fraction in preoxygenation of patients with chronic obstructive pulmonary disease]. AB - OBJECTIVES: To compare the rate of preoxygenation before induction of anesthesia in patients with no lung disease and in patients with chronic obstructive pulmonary disease (COPD). PATIENTS AND METHODS: End-tidal fractional oxygen concentration (FEO2) was monitored using a paramagnetic oxygen analyzer, during a 5 minute-period of preoxygenation (tidal breathing of 100% oxygen) in 16 control patients (control group) and in 15 patients with COPD. COPD was defined and its severity was characterized by clinical criteria and by respiratory functional tests. FEO2 increase was compared between groups using Anova. RESULTS: The increase in FEO2 was slower in the COPD group than in control group (p < 0.05). After 2 and 3 minutes of preoxygenation, FEO2 was significantly lower in COPD group as compared to control group, but was not different at 5 minutes. Mean time to reach a FEO2 equal to 0.90 was significantly longer in COPD than in control group (COPD: 261 +/- 130 s; control: 165 +/- 90 s, p < 0.05). SpO2 measured during room air breathing was moderately lower in COPD group, but this difference was no more significant after 30 s of preoxygenation (SpO2 after 30 s: control: 98.8 +/- 1.0%; COPD: 98.2 +/- 1.9%, NS). CONCLUSION: These results suggest that preoxygenation monitoring may be useful in patients with COPD, to ensure adequate preoxygenation is achieved. PMID- 11878117 TI - [Survey of anesthesia practice in Morocco]. AB - OBJECTIVE: To estimate qualitatively and quantitatively the anaesthetic activity in Morocco to identify of the priorities and the norm to recommend. STUDY DESIGN: The investigation was realised from July 1999 to December 1999. It lasted seven days from Tuesday 8 PM to the following Tuesday 8 PM. A questionnaire was addressed to each hospital. One responsible has been chosen for each city. Data has been processed using the programming languages Microsoft Visual Basic. SETTING: All Moroccan's hospitals practicing anaesthesia: university hospitals (UH), public hospitals (PH), and private sector (PS). PATIENTS: All anaesthetic procedures realised by an anaesthetists; anaesthesia in medical office or dental surgery was excluded. RESULTS: An inventory of 2,630 anaesthesia was made. Anaesthesia distribution according to the type of hospitals was the same; PH = 35.7%, UH = 34.3% and PS = 29.96%. The preanaesthetic consultation was achieved in 47% of cases. Premedication was done in 28.5% of cases. General anaesthesia represented 3/4 of total anaesthesia. Thiopental was the drug the more frequently used. Gallamine and pethidine were still used in PH. Locoregional anaesthesia was realised in only 15% of cases. Electrocardioscopic surveillance not constant (65%), capnography almost absent in PH, pulse oximetry used in UH and PS. Ambulatory anaesthesia was used in 20% and emergency surgery in 30% of cases. CONCLUSION: Questioning about the Moroccan practice of anaesthesia is raised by this survey mostly because of drugs and monitoring use. PMID- 11878118 TI - [Antipublic antibodies and pregnancy: use of iron sucrose in autologous blood donation with cryopreservation]. AB - An autologous blood donation with cryopreservation in a pregnant woman with natural antibody against a high frequency alloantigen is reported. A natural anti Gerbich antibody and a rare erythrocyte phenotype at high risk of polyimmunization was discovered during the third month of pregnancy. This leads to recommend the constitution of an autologous blood reserve. Before first sampling a moderate iron deficiency anaemia (10.3 g.dL-1) was treated with 600 mg of intravenous iron sucrose. Four blood packs of 350 mL were taken; after every sampling 200 mg of iron sucrose were injected intravenously. No maternal or foetal adverse effects occurred. Five weeks before delivery, an autologous blood reserve consisting in 4 cryopreserved red cells packs and 4 fresh frozen plasma was constituted. Epidural analgesia was used for delivery. No haemorrhage occurred. The reserve was not used and remained available for future use (one year for fresh frozen plasma and without limit for red cells). PMID- 11878119 TI - [Narcolepsy and epidural obstetrical analgesia ]. AB - We report the case of a parturient with a history of narcolepsy that delivered under epidural anaesthesia without any complication. Patients with narcolepsy might be at risk of delayed recovery or of apneic episodes after general anaesthesia while narcolepsy-cataplexy fits might be more frequent during regional anaesthesia. Avoiding sedative premedication and continuing medical therapy on the day of surgery are recommended to avoid these complications. PMID- 11878120 TI - [Pneumomediastinum as a complication of recreational ecstasy use]. AB - Ecstasy recreational use dramatically increased during the last decade in France. Multiples complications associated with ecstasy consumption have been reported. We describe an exceptional case of pneumomediastinum related to ecstasy recreational use. The patient was a 19-year old patient whose medical history was significant for asthma and was a moderate tobacco smoker. He reported regular ecstasy recreational use. He presented at the emergency unit for chest pain and cough, 48 hours after the ingestion of three tablets of ecstasy and the inhalation of three other tablets. Blood pressure was 90/60 mmHg, pulse rate was 120 per minute, respiratory rate was 32 per minute and pulse oximetry was 93%. Neck emphysema was noted. Chest radiograph and scanner diagnosed pneumomediastinum. Outcome was favorable with symptomatic treatment and the patient was discharged home eight days later. Pneumomediastinum is an exceptional complication after ecstasy recreational use. Physician should be aware of the risk of such event. PMID- 11878121 TI - [Difficult intubation in patients with endotracheal prosthesis]. AB - We report a case of two consecutive episodes of difficult intubation in a patient with an endotracheal Dumon's prosthesis inserted two years before. Despite several preanaesthetic visits and ENT examination, the absence of recollection by the patient and the lack of information in her previous medical records led to the impossibility to introduce a normal tube into her trachea during two consecutive anaesthetic procedures. This case points out the limits of preanaesthetic visits. It allows to remind special measures that must be taken in patients having tracheal prosthesis and scheduled for anaesthesia and surgery. PMID- 11878122 TI - [Factors associated with blood transfusion during anesthesia for scheduled hip or knee arthroplasty in France]. AB - GOAL OF THE STUDY: To determine over a whole country what are the factors associated with an intraoperative homologous blood transfusion and with the use of autologous techniques (preoperative autologous blood donation: PABD; acute normovolemic hemodilution: ANVH; intraoperative red cell salvage: IRCS). STUDY DESIGN: National enquiry using a large representative sample (3 days of anaesthesia in France). METHODS: Univariate followed by multivariate analyses of data gathered in 1996 during the survey leaded by the French society of anaesthesia and intensive care (Sfar) and corresponding to 884 scheduled hip and knee prosthesis surgical procedures. RESULTS: Factors associated with a decreased use of PABD programme were: 1--old age and high ASA physical status; 2- procedures of short duration. By contrast, an increased use of PABD was associated with anaesthetics in which a closed circuit had been used. Except for a significant association with increasing age and with absence of PABD used, no additional factor was found to be linked with ANVH. No factor among those studied was found related to the use of IRCS. Homologous blood transfusion was more frequently used in ASA > or = 3 patients, in long duration surgeries while its use was decreased in patients with PABD (odds ratio--for reduction by PABD: 4.4 [95% confidence interval: 2.2-8.8]). Homologous blood transfusion was not related to the use of ANVH or IRCS. CONCLUSION: These data obtained from a large national survey confirm previously published studies and meta-analyses and are in agreement with current recommendations. An unexpected relation between PABD and closed circuit anaesthesia has been found. PMID- 11878123 TI - [Cardiac arrest in a traumatized child from an unusual cause: atlanto-occipital luxation]. AB - We report the case of a trauma child who developed a cardiac arrest due to atlanto-occipital luxation of the cervical spine. The occurrence of a rapidly reversible cardiac arrest in a trauma patient should alert physicians about cervical trauma. Adequate resuscitation of these patients require endotracheal intubation with concomitant full immobilisation of the cervical spine, plasma expansion to prevent arterial hypotension and immobilisation of the cervical spine during transport. Thorough application of these resuscitation techniques should increase the survival rate on admission to trauma centres of paediatric patients presenting with such a severe condition. Nevertheless, atlanto-occipital luxation is a major cause of paediatric cervical trauma mortality and our patient did not survive this condition. PMID- 11878124 TI - [Gas embolism and flexible Ecoflac type bag]. AB - Gas embolism at the end of infusion is a well known hazard, that should have disappeared with the use of flexible bags. However, some cases have been reported after pressure infusion. This experimental study evaluates the risk for gas embolism with Ecoflac type flexible bags. These bags are safe under normal pressure infusion conditions with a pneumatic sleeve, because of their texture and pliability; indeed, only minimal air volumes could be expelled, without any risk even in children. However, to be on the safe side, the manufacturer recommends to expel any residual air before pressure administration. This recommendation applies to any bag containing residual air, and since many people are not aware of this, it is rarely put into practice. PMID- 11878125 TI - [Strategies diagnosis of polytraumatized adult patients with coma]. AB - OBJECTIVE: To review the diagnostic strategy of management of multiple trauma patient during the first hours. DATA SOURCES: Extraction from Pubmed database of French and English articles on the management of multiple trauma patient published for ten years. DATA SELECTION: The collected articles were reviewed and selected according to their quality and originality. The more recent data were selected. DATA SYNTHESIS: The first hours of management of multiple trauma patients are a particular challenge. The first dilemma is to drive the patient toward an adequate structure. In case of poor haemodynamic tolerance, the patient will be drive in the nearest hospital. When haemodynamic parameters are restored, multiple trauma patient has to be receive in a high level hospital by a trained medical team with an anesthesiologist, intensivist, neurosurgeon, general surgeon and radiologist. The initial assessment may have two priorities: quality and speed. The total body CT scan is actually the answer to these priorities. PMID- 11878126 TI - [Risk associated with thrombophilia and pregnancy]. PMID- 11878127 TI - [Acute pancreatitis secondary to administration or norfloxacin]. PMID- 11878128 TI - [Anesthesia apparatus: habits are no gauge of safety]. PMID- 11878129 TI - Atopy and helminths. AB - In this article, the authors review current and latest evidence linking helminth infections and the development of atopy. Although there is intense ongoing investigation and debate on this issue, the review of experimental, clinical and epidemiological data apparently shows that helminth infections can have beneficial aspects in regard to the pathogenesis of atopy and allergic diseases. Despite the fact that simplistic views are not recommended, it seems that polyclonal IgE production and mainly the stimulation of host immunoregulatory networks leading to synthesis of anti-inflammatory cytokines (IL-10, TGF-beta and others) can provide new insights into how mechanisms that helminths have developed throughout their evolution and that are useful for parasite evasion and persistence, could also be used in humans in order to provide new approaches in atopy prevention. PMID- 11878130 TI - [Evolution of eosinophilia during the pollen season among the general population: a way of determining new sensitiziations]. AB - The purpose of this paper was to verify the effect of pollen peaks on blood eosinophilia in an all and sundry population, including allergic as well as non allergic subjects, so that we can open up new horizons in the understanding and prevention of pollinosis. Daily eosinophilia counts of hospital patients were measured at the time of a blood checkup (1996-1998), and divided into six classes. Those data were compared to daily pollen counts of twelve taxa, coming from the Hirst trap of Dijon (France). An eosinophilia increase occurred when hazel, hornebeam, birch, oak, grasses, ragweed and plantain were present in high concentration. In other cases, only simultaneous presence of several taxa seemed to play a part, because of cross-reactivity or polysensitization. Lastly, Cupressaceae-Taxaceae and ragweed were seen as increasing eosinophilia in seemingly non allergic people. The analysis of eosinophilia in the general population was able to reveal potential allergic patients and potential allergic diseases. PMID- 11878131 TI - [The oral mucosa and delayed hypersensitivity]. AB - Buccal mucosal allergies are rare because the conditions for sensitization are less favourable in the buccal environment. Clinical presentation is dominated by subjective manifestations (hardening, hypersialorrhea, "burning mouth" syndrome). Extradental etiologies are linked with cosmetic and medicinal topics, as well as dental etiologies caused by metals, alloys of mercury and resins. PMID- 11878132 TI - Respiratory infections in paediatrics: recent findings and prospects. PMID- 11878133 TI - Laboratory diagnosis of hymenoptera venom allergy: comparative study between specific IgE, western blot and allergen leukocyte stimulation (CAST). AB - Allergy to hymenoptera venom is a classical IgE mediated disease with a potentially fatal course. Specific venom immunotherapy (SIT) is the most effective mean of treating this serious condition, after the diagnosis has been clearly established by a clinical history, in-vivo and in vitro tests. We have evaluated the usefulness of a cellular test (CAST) which is a recently developed ELISA method based on the evaluation of sulfidoleukotriene secretion by leukocytes stimulated with specific antigen. We also evaluated the correlation of CAST with skin tests, specific IgE (sIgE) and western blot for sIgE for hymenoptera venom sIgE. We have included in this study 14 patients, with a clinical history suggestive of hymenoptera venom allergy. None of them had previously been subjected to immunotherapy. A good correlation was obtained between skin tests, sIgE and western blot. However, there was no correlation between these methods and CAST. We conclude that the positivity of CAST method raises some questions about other mechanisms, which maybe non-IgE dependent. Although the number of patients in this study is quite small, the immunoblot analysis may be a valuable additional method in insect venom allergy. PMID- 11878134 TI - [Exercise intolerance and muscular dystrophies]. PMID- 11878136 TI - [Prevalence of lower urinary tract disorders and incontinence in Auxerre. French segment of the UrEpik study]. AB - OBJECTIVES: To estimate the prevalence of low urinary tract symptoms (LUTS) and urinary incontinence among men and women aged 40 to 79 years. METHODS: A postal survey was carried out in a community-dwelling random sample, in Auxerre, France. LUTS were assessed using the International Prostate Symptom Score (IPSS). Urinary incontinence (UI) was assessed using a score based upon four questions. RESULTS: 1216 men and 591 women participated in the survey. 19.2% of men and 13.7% of women reported moderate to severe LUTS (IPSS > 7). 73% of men and 29% of women suffered from UI. Among men and women reporting severe symptoms of UI (0.4 and 5.4% respectively), one third had been prescribed medications for urinary disorders within the previous six months. CONCLUSION: This survey confirms the high prevalence of LUTS and urinary incontinence among a community-dwelling sample of men and women, and highlights the need for management of these disorders. PMID- 11878135 TI - [Muscular exercise intolerance syndrome in Becker muscular dystrophy]. AB - OBJECTIVE: To determine whether exertional muscle pain syndrome (EMP) is a benign phenotype or an early stage of Becker-type muscular dystrophy (BMD). METHODS: Muscle dystrophin analysis led to the diagnosis of BMD in 6 patients complaining of EMP. RESULTS: Three patients had a history of X-linked inheritance and age at clinical onset was 4 to 11 years in five, and one patient had a later onset aged 23. Pseudohypertrophy of the calf muscles was absent in one patient, but all had experienced mild (5/6) pelvic weakness and (or) atrophy one to 17 years after the onset. High serum CK level was present (X 14). Normal anti-dystrophin immunostaining in two cases did not rule out the diagnosis that was only made made by Western blot analysis or genetic studies. All exhibited in-frame deletions (exons 45-48) within the dystrophin gene. CONCLUSION: The 36 patients with BMD-EMP analysed in the literature, exhibited different deletions and no worsening in 66.7% of cases. Western blot was more precise than immunolabelling with 96.8% positivity versus 70.5%. Dystrophin analysis by Western blot and (or) DNA analysis should be included in the evaluation of patients with EMP syndrome without deficient muscle energy metabolism, particularly those with pseudohypertrophy of the calf muscles or high serum CK levels. PMID- 11878137 TI - [Anterior pituitary insufficiency caused by bilateral carotid artery aneurysms]. AB - INTRODUCTION: The frequent etiology of panhypopituitarism is a hypopituitary adenoma. An exceptional cause, with less than 40 cases in the literature, is intra-sellar carotid aneurism. OBSERVATION: A 55 year-old woman was treated for panhypopituitarism related to bilateral aneurism of the carotid arteries. Hormone exploration revealed anterior pituitary deficiency involving all the axes except the corticotrophin and prolactin functions. Pituitary MRI showed an atrophic pituitary, compressed by two calcified aneurismal formations located on the terminals of two internal, partially thrombotic, carotid arteries. Surgical treatment was not envisaged 12 years later. Replacement therapy for life was initiated. COMMENTS: The mechanisms explaining the occurrence of panhypopituitarism secondary to intra-cranial aneurism are hypothetical: compressive phenomena, ischemia and apoplexy of a pituitary adenoma related to the aneurism. PMID- 11878138 TI - [Malignant histiocytofibroma of the small intestine in a young immune deficient patient]. AB - INTRODUCTION: The visceral localization of malignant fibrous hystiocytoma is rare. It provokes polymorphous, rarely specific symptoms. Although muscular or cutaneous forms of the disease are frequent in the elderly, the visceral form predominates in young immunodepressed patients. OBSERVATION: An 18 year-old, HIV seropositive man was hospitalised for chronic abdominal pain and transit disorders. Biological examination revealed eosinophilia and imaging showed a thickening of the intestinal wall of the right iliac fossa. Surgery revealed an intrinsic tumour on the intestinal wall. The anatomopathological exploration evoked a malignant storiform histiocytoma. Despite exeresis, the tumour relapsed a few months after surgery, leading to the death of the patient. COMMENTS: This case report can be added to those of atypical localization of rare tumors in immunodepressed and young patients. The localization in the digestive tube is rare and the cases reported referred to tumors of the colon or stomach. Our case report confirms the observations of the concomitance of malignant fibrous histiocytoma and eosinophilia. The association with bacillary angiomatosic lesions is surprising and has yet to be explained. PMID- 11878139 TI - [Mirizzi syndrome]. PMID- 11878140 TI - [Silent myocardial ischemia in type 2 diabetes. Predictive value of intima-media thickness]. AB - CONTEXT: The carotid intima-media thickness (CIT) is correlated with the prevalence of cardiovascular diseases. We studied the relationship between the thickness of the carotid intima-media and the existence of myocardial ischemia, detected by scintigraphy in a population of type 2 diabetic patients without coronary antecedents. METHODS: We conducted repeated Doppler measurements of the CIT of primitive carotid arteries in 52 diabetic patients aged 49 to 75. All these asymptomatic patients, without coronary antecedents, diabetic for more than 10 years and exhibiting at least one cardiovascular risk factor, had undergone myocardial scintigraphy. RESULTS: The CIT was greater in the group with positive scintigraphies and is the best predictive factor of the presence of ischemia. For a CIT value < 0.55 mm, the negative predictive value of the CIT was of 77% with 80% sensitivity; in non-smokers the predictive negative value increased to 92% with 95% sensitivity. CONCLUSIONS: Reliable and reproducible, measurement of CIT correlates well with myocardial scintigraphy and could be an interesting alternative screening policy in asymptomatic, type 2 diabetic patients. PMID- 11878141 TI - [Indications for medical lasers in dermatology]. AB - FOUR CATEGORIES OF LASERS ARE USED IN DERMATOLOGY: These are vascular, depigmentation, depilatory or resurfacing or vaporization lasers. Today, there are more potential or suggested indications than good methodological studies that confirm these indications. Nevertheless, there are indisputable indications for these lasers. VASCULAR LASERS: They are indicated in the treatment of capillary nevus in adults and children and, with pulsed dye lasers, infants can be treated within the first weeks of life. Another indication for vascular lasers is treatment of Stage II rosacea, i.e., at the stage of telangiectasic erythrosis or couperose. Other indications include radiodermatitis, ulcerated hemangioma and erythrosis of the neck. DEPILATORY LASERS: Treatment of patients with pale phototype and dark hairs appears possible. However, around 4 to 6 sessions are required to obtain significant lasting hair removal. DEPIGMENTATION LASERS: The best indications are the removal of tattoos, Ota's nevus and, to a lesser degree, liver spots and Becker's nevus. Melasma and chloasma are not indications or exeresis of nevo-cellular nevi using this technique, since no histological control is possible. PULSED VAPORIZATION LASERS (CO2 OR ERBIUM LASER): They permit dermabrasion in the treatment of verrucous harmatoma, extensive benign superficial dermo-epidermal lesions and the esthetic treatment of non-muscular wrinkles, i.e., excepting wrinkles of the forehead and nasal sulcus. Continuous CO2 lasers destroy small dermo-epidermal lesions. They are particularly indicated for profuse lesions, in which there is a risk of hemorrhage or when direct contact should be avoided because of potential HIV infection. CONCLUSION: There are many potential indications, but a consensus has only been reached on those mentioned. The others remain to be confirmed. PMID- 11878143 TI - [The MISA study: what about benzene?]. PMID- 11878142 TI - [Frequency of persistent or transitory serologic negative values in infants with congenital toxoplasmosis. Experience of the Reims Toxoplasmosis Group (1980 1997)]. PMID- 11878144 TI - [Does the Vatican kill with the waves? The difficult question of data interpretation]. PMID- 11878145 TI - [Is mammography screening efficient? Let's open the debate]. PMID- 11878146 TI - [Improving the early diagnosis: that is the challenge]. PMID- 11878147 TI - [The Ministry's Working Group on the Vatican Radio Station and the case of the "midwife toad"]. PMID- 11878148 TI - [The Vatican Radio Station and leukemia: an alarm without response]. PMID- 11878149 TI - [Critical considerations about the Ministry's Report on the Vatican Radio Station]. PMID- 11878150 TI - [Leukemia mortality and incidence of infantile leukemia near the Vatican Radio Station of Rome]. AB - Some recent epidemiological studies suggest an association between lymphatic and haematopoietic cancers and residential exposure to high frequency electromagnetic fields (100 kHz-300 GHz) generated by radio and television transmitters. Vatican Radio, a very powerful radio station transmitting all over the world (up to 600 kW) is located in Santa Maria di Galeria, in the northern suburbs of the city of Rome. Electric field measurements in the proximity of the radio station ranged between 1.5 and 25 V/m. In the 10 km area around the station, with 60.182 residents (1999), leukaemia mortality among adults (> 14 years, 40 cases) in the period 1987-98 and childhood leukaemia incidence in the period 1987-99 (8 cases) were evaluated. The analysis (Stone's conditional test) was performed computing observed and expected cases (reference: population of Rome) in 5 bands of increasing radius (2 km width). The risk of childhood leukaemia was higher than expected within 6 km from the station (Standardized Incidence Ratio = 217; 95% Confidence Interval 99-405). Stone's test showed a significant decrease in risk with increasing distance both for male adult mortality (p-value = 0.03) and for childhood incidence (p-value = 0.04). A Score test, showed a significant decrease in risk of childhood incidence as function of the distance. The main limitations of this study are the small number of observed cases and the use of distance as a proxy for RF exposure. Further research will require a systematic campaign of electromagnetic field measurements to allow better assessment of the population exposure. PMID- 11878152 TI - [Recovery of the Archives of the Italian Society of Asbestos of Grugliasco, Turin]. PMID- 11878151 TI - [Interpretation problems in the analysis of temporal series of air pollution and health. Part II: significance of outcome variables, relations forms, effect modifiers, and mechanisms of action]. AB - This paper is a commentary to the MISA study (Meta-analysis of Italian studies on short-term effects of air pollution). The interpretation of the relationships observed in the time-series analyses is not straightforward. I will review a number of open questions: the exposure estimates; the meaning of the daily mortality and hospitalisation rates, in terms of impact on life expectancy, incidence of specific diseases, exacerbation of preexisting chronic diseases; the pattern of the estimated dose-response curves; the role of possible effect modifiers; the hypotheses about the underlying mechanisms. These interpretative problems often present overlapping aspects. Some of the pending answers are relevant for policy making. For editorial reasons this review is divided in two parts. A first paper, focusing on the exposure estimates, has been published on the previous issue of this journal. This second part deals with the meaning of the outcome variable, the shape of the dose-response curves, possible effect modifiers and underlying mechanisms. PMID- 11878153 TI - [Analysis of cohort studies subtracting a lag period]. PMID- 11878154 TI - [Predictive value of reactive strips in urinary infection]. AB - The microbiologic method to confirm or not the diagnosis of urinary tract infection is usually the culture and Gram-stain of the clean-catch midstream urine. Nevertheless, this is a costly and time-consuming method. We tried to evaluate the positive and negative predictive value of the nitrite and leucocyte esterase reactive strips as an alternative method. When both the two parameters are negative it is fairly safe to exclude the urinary tract infection. PMID- 11878155 TI - [Antimicrobial susceptibility of Streptococcus pyogenes, Haemophilus influenzae, Streptococcus pneumoniae and Moraxella catarrhalis from community acquired respiratory infections in 2000]. AB - The Viriato Study is a nationwide, multicenter prospective study of the antimicrobial susceptibility of bacterial pathogens commonly associated with community-acquired respiratory tract infections in Portugal. In 2000, 28 laboratories participated in the study with a total of 1071 strains, with testing undertaken in a central laboratory. Of the 213 Streptococcus pyogenes strains isolated from patients with acute tonsillitis, all were susceptible to penicillin, amoxicillin/clavulanate and cefuroxime, but 21.1% were resistant to erythromycin, clarithromycin, azithromycin and 16.4% to tetracycline. From patients with lower respiratory tract infection, 403 strains of Haemophilus influenzae, 366 of Streptococcus pneumoniae and 89 of Moraxella catarrhalis were studied. 13.1% of H. influenzae and 94.4% of M. catarrhalis produced beta lactamase. Among S. pneumoniae isolates, 25.1% were resistant to penicillin (8.8% showing high-level resistance), 14.5% to tetracycline, 12.8% to erythromycin, clarithromycin and azithromycin, and 10.1% to cefuroxime. Overall, penicillin was the most active antimicrobial against S. pyogenes and amoxycillin/clavulanate the most active in vitro simultaneously against H. influenzae, S. pneumoniae and M. catarrhalis isolated from patients with community-acquired LRTI in Portugal. PMID- 11878156 TI - [Primary neoplasm of unknown origin]. AB - Cancers of unknown primary site constitute a heterogeneous group having in common their biological behaviour. We have analysed patients with cancer of unknown primary site admitted to University Hospital of Coimbra/Internal Medicine Service III from 1st January of 1992 to 30th June of 1998. The study included 26 patients corresponding to a prevalence of 2.5% malignant neoplasias; the median age of affected patients was 64 years old with a male-to-female ratio of 4.2:1. The most frequent histological type was well differentiated and moderately well differentiated adenocarcinoma, 46.1% of all cases. Three years global survival rate was 3.9%. The herein-reported prevalence is comparable to that found in recent literature, showing a good diagnostic skill. As expected, the prognosis was poor, revealing the disease's aggressiveness and the lack of efficient therapy in the majority of case. PMID- 11878157 TI - [Anticipation and quality of life in depression: 2 cognitive concepts?]. AB - This study purports to investigate the relation between negative predictions and quality of life occurring in dysthymia and non-melancholic depression according DSM-IV, in the absence of any personality disorders. The relation between anticipation and quality of life (QoL) is also examined. The results suggest that both anticipatory cognitions and the QoL discriminate the two depressive entities and may have some nosological prediction potential. It is curious to observe the similar behaviour of the two concepts, anticipation and QoL, when implemented with the chosen scales. Moreover, their strong mutual correlation suggests a conceptual proximity or even a concurrent criteria validity. In conclusion, the possibility that both concepts belong to a cognitive sphere should not be ruled out. PMID- 11878158 TI - [Prevalence of antibodies against hepatitis A virus in a population from northern Portugal]. AB - AIM: To find the prevalence of antibody to hepatitis A virus in the population of the North of Portugal. MATERIAL AND METHODS: Ten General Practitioners were asked to provide blood samples from patients who would need blood tests for any reason other than acute hepatitis, during January and February 1996. In this way, 381 samples were obtained for assessment of anti hepatitis A virus antibodies using a commercial radioimmunassay ELISA. All subjects gave their informed consent and answered to a protocol regarding age, sex, geographic area, number of people per household and sewage systems. The statistics were performed using SPSS. RESULTS: The 381 subjects were distributed into eight age groups: I (1-4 years)--57; II (5 9 years)--57; III (10-14 years)--26; IV (15-19 years)--41; V (20-29 years)--55; VI (30-39 years)--51; VII (40-49 years)--41; VIII--(equal or more than 50 years)- 53. The prevalence of anti HAV antibodies per group-percentage (number), (confidence intervals), were: I--7.0% (4) (3-17%); II--15.8% (9), (9-27%); III- 26.9% (7) (14-46%); IV--51.2% (21) (37-66%); V--85.5% (47) (74-92%); VI--72.5% (37) (59-83%); VII--87.8% (36) (75-95%); VIII--88.7% (47) (80-93%). CONCLUSION: The comparison with previous data (Lecour et al.) shows improvement in sanitary conditions of population, with associated lower prevalence of anti hepatitis A virus antibody. PMID- 11878160 TI - [Neuroimaging features associated with AIDS]. AB - Due to the nonspecific clinical presentation of Central Nervous System disorders that may come out during the course of Human Immunodeficiency Virus infection, we emphasize the importance of neuroimaging on the differential diagnosis of these situations. Review and illustration of the most typical imaging patterns of the main Central Nervous System disorders in Acquired Immunodeficiency Syndrome. PMID- 11878159 TI - [Psoriatic arthritis]. AB - The Psoriasis Arthritis witch was considered for many years as a late complication of the Psoriasis, is presently known as a chronical systemic disease, resulting w from the coexistence of a chronical evolutive poliarthropaty and a skin disease, the Psoriasis. The authors make a brief revision of the etiopathogeny, clinics and treatment of the Psoriasis arthritis and show a retrospective analysis of the clinical cases of 19 patients with Psoriatic Arthritis followed at the Medicine Consultation Reumathologie. PMID- 11878161 TI - [The discovery of the body]. AB - This paper initially recognises that the human body tends to implicate itself in the current triad in which it gains evidence (body, soul and spirit). Its discovery is, therefore, a difficult cognitive process. In the conveyance of the message that one has from the body to the desideratum of its knowledge, the possibility of discovery fulfills itself in the adventure of its study where religious, artistic and scientific paths distinctively appear as modes of evidence. As discovery is usually entrusted on medicine it is nowadays important that the latter should pronounce itself on acquired knowledge. Health and its re establishment after illness are correlated with the knowledge of the body! In order for the allegations regarding the body's rights over the body to be more than mere phantasmagoric exorcisms, it is necessary to consider the knowledge of the body as an experience, particularly, of existing, of having and of being. PMID- 11878162 TI - [Aggressive angiomyxoma of the vulva]. AB - An unusual myxoid and vascular appearing neoplasm of the vulva, termed an Aggressive Angiomyxoma, is described in two cases report. The clinical presentation and recurrence pattern were typical for previously described neoplasms of the same cellular pattern. The histopathology, difficulties in determining the surgical margins, and the treatment of this tumor are discussed. PMID- 11878163 TI - [Dysgenetic male pseudohermaphroditism]. AB - Dysgenetic male pseudohermaphroditism is the result of a defect of testis development that encompasses a large clinical heterogeneity. It is characterized by bilateral dysgenetic testis, absence of mullerian regression, ambiguous genitalia and/or stigmata of Turner's syndrome in the majority of the cases. Typically, these individuals have either a 46,XY or 45,X/46,XY karyotype. The authors present four cases of dysgenetic male psudohermaphroditism, with ages of diagnosis between 1 month and 17 years old. The first had a male phenotype with stigmata of Turner's syndrome and the others ambiguous genitalia. Two patients were 45,X/46,XY and 45X/47,XYY mosaics and the other two were 46,XY. Gonadal karyotyping showed mosaicism (45,X/46,XY) in all four cases. In the first case was programed orquidectomy; all the others assigned a male gender, with regular follow-up until the puberty. PMID- 11878164 TI - [Intracavitary heart metastasis of testicular embryonic tumor]. AB - Cardiac intracavitary metastasis are very uncommon. A 19 years old patient with an embryonic tumour of the testes extending into the right ventricle, which manifested as heart failure, is presented. To our knowledge, this is the first case report of a teratoma with carcinoma of the testis involving the right ventricle. PMID- 11878165 TI - [Crohn disease with unusual cutaneous expression]. AB - Perianal fistula is a frequent complication of Crohn's disease. The diagnosis of the later may be difficult if cutaneous disease is dominant, and sometimes an important clinical challenge. The authors report a case of a 34 year old male complaining of painful perianal nodules for the three last years. Cutaneous histopathological examination was inespecific and colonoscopy normal. Pelvic CT scan and fistulography showed an intercommunicated fistular system invading both isquiorectal fossae, without intestinal communication. Repetition of colonoscopy showed normal mucosa, but in two of the biopsy samples noncaseous granulomas were found at the submucosa. The authors explain their main difficulties to reach the diagnosis of Crohn's disease and discuss the investigation and the evolution of the patient. PMID- 11878166 TI - [Asymptomatic hepatitis induced by propylthiouracil]. AB - Propylthiouracil (PTU) is widely used to treat patients with hyperthyroidism. In rare cases this drug has been found to have toxic effects on the liver. We describe the second assymptomatic case, reported in the literature, of PTU induced hepatotoxicity. Sixteen cases have been published so far, all with clinical manifestations. A 20-year-old female patient developed an elevation of AST, ALT and GGT levels after 5 months of PTU (50 mg tid) therapy for Graves disease with hyperthyroidism. The patient showed normal levels of liver aminotransferases before starting PTU. She had no risk factors for or clinical evidence of pre-existing liver disease. She didn't take any regular medication. After stopping PTU, the plasma levels of hepatic enzymes normalised within 6 weeks. Thirteen days after the reinstitution of PTU (150 mg tid), because of the recurrence of hyperthyroidism, another episode of assymptomatic hepatotoxicity was documented. AST, ALT, GGT and AP levels rose again, but bilirrubin levels remained normal, which represents a very unusual laboratory constellation in this kind of patient. The serology for hepatitis A, B, C and for EBV and CMV was negative. After stopping PTU, methimazole (MMI) was started and the liver enzymes normalised within two weeks. After 12 months of MMI therapy, the patient is assymptomatic, with normal liver enzyme levels and normal thyroid function tests. This case is the second to be described where the diagnosis was confirmed by the rechallenge of PTU, being this a major diagnostic criteria. Despite its rarity the disease should be suspected in any patient receiving PTU in whom clinical or laboratorial evidence of hepatocellular injury develops. The drug should be discontinued immediately when the hepatic injury is detected. Recovery is usually complete after the withdrawal of the drug, but there were al least, three fatalities among the 17 cases described. PMID- 11878167 TI - [The Portuguese version of the 20-item Toronto Alexithymia Scale -- I. Linguistic adaptation, semantic validation, and reliability study]. AB - BACKGROUND: The Toronto Alexithymia Scale, namely in its revised version (TAS 20), is nowadays widely used and certainly the most well validated measure of alexithymia; allowing to recognize three distinct aspects of the construct. OBJECTIVE: The aim of the present study is to assess reliability and validity of a Portuguese translation; proceeding in this first part, to the semantic validation and confirmatory structural analysis of the questionnaire, as well as to the analysis of the internal coherence of this structure, and of its reliability in what concerns time stability. Just to later examine validity related aspects as described in a second paper. METHODS: Fist of all translation and substantiation were properly made as described according with commonly used procedures. Thus resulting in a questionnaire subsequently applied to three distinct populations: university students (N = 468), outpatients attending a routine general practice consultation (N = 50), and patients with inflammatory bowel disease (N = 74). RESULTS: Despite some traceable instability while replicating the original model, confirmatory factorial analysis somehow allowed disclosing the three originally described factors. Nevertheless also resulted some differences susceptible to later being discerned among the factors in what concerns their internal coherence: the questionnaire as a whole is highly reliable, and the same goes for factor 1, in any of the studied samples; less steady results among different groups were found regarding factors 2 and 3. However the test-retest stability brought to evidence, in any of the considered groups, an excellent reliability in all aspects: global score and integrating factors. CONCLUSION: With these sound psychometric properties of the instrument under scrutiny, the observed cross-cultural variations result as minor, while also considering the aimed comparability in the context of a vast amount of alexithymia studies carried out all over the world. PMID- 11878168 TI - [Allocation at central and university in Portugal -- severe penalization of research]. PMID- 11878169 TI - Use of chemically extracted muscle grafts to repair extended nerve defects in rats. AB - Nerve regeneration, measured as axonal outgrowth, Schwann cell migration, macrophage invasion, and neovascularisation, was compared after repair of a 15 mm gap in rats' sciatic nerves using autologous muscle grafts made acellular either by freezing and thawing or by chemical extraction. Both extracted and freeze thawed acellular muscle grafts could be used to bridge the defect. However, axons and Schwann cells, as shown by immunohistochemical staining for neurofilaments and S-100 protein, respectively, grew faster into the extracted muscle grafts than into the freeze-thawed acellular muscle grafts and somewhat more axons were observed in the former graft. There were no significant differences between the two graft types with respect to neovascularisation as showed by staining for endothelial alkaline phosphatase, and limited differences concerning invasion of macrophages (ED1 and ED2) as detected by immunocytochemistry. The results showed that chemically extracted muscle grafts could be used to bridge an extended nerve defect and that such grafts in some aspects were superior to freeze-thawed muscle grafts for extended gaps. PMID- 11878170 TI - Different effect on axonal outgrowth of application of non-absorbable or absorbable tubes around a nerve repair. AB - We studied regeneration distance of rat sciatic nerve, with the sensory pinch reflex test and immunocytochemical staining for neurofilaments, four to 21 days after transsection, repair, and enclosure of the repair site in either a non absorbable silicone tube or an absorbable polyglycolic acid (PGA) tube. The size of both tube-types was carefully selected so that they did not compress the repaired nerve. The opposite nerve was repaired and not inserted in a tube (control). The regeneration distances in repaired nerves enclosed in silicone tube were significantly longer than the control side at all time points, a result not seen when PGA tube was used. The number of proliferating non-neuronal cells (incorporation of 5-bromodeoxyuridine (BrdU)) was studied just proximal to the site of nerve repair after six days. Numerous stained cells were seen, but there where no significant differences between the groups. We conclude that outgrowth of sensory axons after transsection and repair of rat sciatic nerve with sutures can be increased by enclosing the site of repair in a silicone tube but not in a PGA tube. The effect is probably not related to the number of proliferative non neuronal cells. PMID- 11878171 TI - Plexus avulsion and spinal cord injury increase the serum concentration of S-100 protein: an experimental study in rats. AB - The possibility of using the presence of the glial-cell-derived protein S-100 in serum as a marker for neuronal damage caused by spinal cord injury and plexus avulsion injury was investigated in 144 adult rats. After a spinal cord injury had been induced at the thoracic level or a plexus avulsion injury at the lumbar level, blood samples were taken and analysed for S-100 protein by a monoclonal two-site immunoluminometric assay. The two types of neurotrauma changed the kinetics of serum S-100 in different ways. After spinal cord injury it rapidly increased and within 72 hours had reached a concentration about 5 times that of the control animals. Three peak concentrations occurred at 3, 12, and 72 hours, respectively, and differed significantly from those of the control group (p < 0.05). After six days the values had returned to normal. After lumbar plexus injury alone there was no significant increase in the concentration of S-100. These results suggest that the concentration of S-100 protein in serum may be used as an early diagnostic tool for detecting neuronal damage caused by spinal cord injury or plexus avulsion associated with damage to the root entry zone. PMID- 11878172 TI - Effects of gradual bone lengthening on the rabbit tibial nerve. AB - Little is known about the effect of gradual bone lengthening on peripheral nerves. In the present study, an external fixation device was applied to the rabbit tibia, which was then divided. After seven days, the tibia was subjected to 0.7 mm/day callus distraction for periods of up to one month. The tibial nerve was fixed in glutaraldehyde and plastic sections were cut in longitudinal and transverse planes for light and electron microscopy. Light microscopy showed a 64% increase in the gap length at the node of Ranvier in myelinated axons from the experimental side compared with the control side. The cross-sectional area of the non-myelinated axons was not altered significantly. We conclude that gradual stretching of the nerve elongates the nerve fibres at least at the region of the nodes, perhaps a point of least resistance. Diameters of fibres seem to be held more constant during the lengthening procedure. PMID- 11878173 TI - A biodegradable bovine collagen membrane as a dermal template for human in vivo wound healing. AB - A bovine collagen membrane was used as a template for dermal regeneration in human full thickness wounds. Healing was allowed for 7, 21, or 42 days. The formation of neodermis, basement membrane, and terminal differentiation were assessed histologically and immunohistochemically. The collagen template was neovascularised within 7 days, and from day 21 small vessels were detected throughout the transplanted area. The procollagen content decreased whereas the number of fibroblasts increased with time. Collagen type IV was not detected after 7 days but was deposited with time from the wound edges and inwards over the transplanted area. Re-epithelialisation was complete at day 7 and terminal differentiation was similar to normal human skin from day 21. We have shown the time course of dermal and epidermal healing with the aid of a ready-to-use biodegradable collagen membrane. This material may be used as a true dermal template-because of the evidence of dermal regeneration and, in addition, its availability and ease of handling. PMID- 11878174 TI - Maxillary development and dental occlusion in patients with unilateral cleft lip and palate after combined velar closure and lip-nose repair at different ages. AB - Maxillary morphology and occlusal development were studied after simultaneous velar closure and lip/nose reconstruction in patients with unilateral cleft lip and palate. Fifty-two Brazilian patients were divided into three groups according to the age at which they had had the one-stage operation (mean ages: 8, 18, and 77.5 months). They were compared with 30 similar white patients who had been operated on with a corresponding method, but where surgery had been carried out in three different stages. In general, differences in outcome between the groups were attributed to racial differences in facial morphology. The combined operation did not affect the transverse development or the overall occlusion and only slightly influenced the morphology of the maxillary incisor region. The palatal cleft width reduced significantly (p < 0.001) after the combined procedure. However, the potential for this reduction seemed to be less when patients were operated on after their first year of life. PMID- 11878175 TI - Adult skeletal profile in isolated cleft palate: a comparison of the von Langenbeck and Wardill procedures for primary repair of the palate. AB - Sixty-four adult patients operated on for isolated cleft palate were evaluated with regard to facial skeletal morphology using conventional radio-cephalometry. Dental occlusion was assessed clinically. Forty-two had had a von Langenbeck repair at the age of 7 months and 22 a Wardill repair at 18 months. The mean error of the method was 0.7 degree for angular, and 0.9 mm for linear, measurements. The group with clefts had less maxillary prognathism (s-n-ss), more maxillary inclination (NSL/NL), more retroclined lower incisors (ILI/ML), and shorter total and upper facial heights (n-gn, n-sp) compared with the reference group. Multiple regression analysis was used to evaluate differences between the two treatment regimens. Explanatory variables in addition to surgical technique were sex, severity of cleft, and presence of a velopharyngeal flap. Only one variable, lower incisor inclination (ILI/ML), was different for the two regimens. Ten (24%) in the von Langenbeck group had a lateral cross-bite compared with one (5%) in the Wardill group. Other variables in a multivariate regression analysis were affected by sex and severity of cleft to various degrees. This study showed no obvious differences in facial skeletal morphology that could be attributed to surgical technique. Factors other than technique, including sex, age, and severity of cleft merit attention. PMID- 11878176 TI - Patients' satisfaction with breast reconstruction and reduction mammaplasty. AB - Patients' satisfaction is widely recognised as an important measure of the quality of care. This study measured patients' satisfaction with the outcome of breast reconstruction surgery and reduction mammaplasty and with the treatment received in hospital. A questionnaire about patients' reasons for having the operation and postoperative satisfaction was therefore sent to 137 patients after breast reconstruction and 142 who had had reduction mammaplasty. The response rate in the former group was 84% and in the latter 65%. In both groups of patients the main reasons for having the operation were difficulties with physical activities and in finding clothes to fit. Other factors were poor self esteem and problems with body image. A total of 69% of the patients who had had reconstructions referred to problems with wearing an external prosthesis. In the reduction mammaplasty group one of the most important reasons for having the operation was pain, in particular neck, shoulder, and back pain. In the reconstruction group 94%, and in the reduction group 88%, thought the outcome of the operation was good or very good. They had no regrets about their decision to seek surgical treatment, even when the aesthetic outcome fell short of their expectations. The patients were generally satisfied with the treatment they had received. In the reconstruction group 97%, and in the reduction group 96%, of patients felt satisfied with their overall decision to have the operation. Our results confirm that these surgical procedures help to improve patients' quality of life. Among the patients who had had reconstructions, the resolved problems with body image were considered to be the main benefit of the operation. In the reduction group the main benefits were reduced neck, shoulder, and back pain. In both groups 91% of patients would have recommended the operation to a friend under similar circumstances. In the light of this evidence doctors should more readily recommend reconstruction to patients after mastectomy and reduction to women with heavy breasts. PMID- 11878177 TI - Long-term results of arthrodesis of the carpometacarpal joint of the thumb. AB - We reviewed 48 arthrodeses of the carpometacarpal joint of the thumb in 39 patients being treated for osteoarthritis. The mean follow-up period was 90 months (range 14 months-17 years). In 28 (58%) hands pain relief was excellent and in 6 (12%) good. Morbility was significantly reduced (p < 0.002). Mean key pinch (52.9 N) and grip strength (219.5 N) were comparable with the unoperated hand (51.0 and 239.1 N, respectively). Hand function was near normal. In 29 hands an additional operation was needed to remove the osteosynthesis material; 14 hands failed to unite. Osteoarthritis of the scaphotrapezial joint increased significantly faster in the operated hands than in the unoperated ones (p < 0.0001). A soft tissue procedure (such as flexor carpi radialis interposition arthroplasty) is advised instead of arthrodesis, because of the large number of reoperations, based on non-union and removal of the osteosynthesis material. From a survey of published papers we conclude that arthrodesis of the carpometacarpal joint of the thumb can best be done by stapling and bone grafting. PMID- 11878178 TI - Reduction in the need for operation after conservative treatment of osteoarthritis of the first carpometacarpal joint: a seven year prospective study. AB - The effect of occupational therapy for patients awaiting surgery for isolated osteoarthritis of the carpometacarpal joint of the thumb was assessed. Thirty three patients awaiting joint replacement because of pain were randomised into three groups. One group was treated with technical accessories, two other groups had in addition one of two types of splints, and all patients received extensive advice on how to accommodate activities of daily living. They all had an initial seven months' trial on this regimen at which time 23/33 (70%) no longer required an operation. During the following seven years four patients died, but only two of the remaining 19 patients wanted an operation. We therefore recommend that patients with osteoarthritis of the carpometacarpal joint of the thumb are offered a similar programme in addition to access to accessories and splints preoperatively. PMID- 11878179 TI - Cleft palate in spondyloepiphyseal dysplasia congenita: case reports. AB - Cleft palate is one of the common features of spondyloepiphyseal dysplasia congenita (SEDC). However, there are few clinical data about cleft palate in SEDC. We report four patients with cleft palate and SEDC including two with overt cleft palate and two with submucous cleft palate. Our results suggested that SEDC associated with cleft palate should be treated in the same way as solitary cleft palate, and submucous cleft palate may be more common in patients with SEDC than previously appreciated. PMID- 11878180 TI - Reconstruction of a subtotally amputated auricle: a case report. AB - A 42-year-old woman presented with a subtotal amputation of the left auricle except for the helical skin. The treatment was by primary suture with debridement of some margins, and the auricle healed well. The fact that the treatment was successful without microvascular anastomoses is important, considering the anatomical features of the auricular vascular networks. PMID- 11878181 TI - Magnetic resonance imaging of a tiny glomus tumour of the fingertip: a case report. AB - Magnetic resonance imaging (MRI) was used for a patient with severe pain in the fourth finger of her left hand when other imaging modalities had failed. Magnetic resonance scan with short T1 inversion recovery (STIR) sequences and gadolonium enhancement successfully aided in the diagnosis of a tiny glomus tumour. PMID- 11878182 TI - Tumoral calcinosis in both index fingers: a case report. AB - Tumoral calcinosis is an uncommon disease of unknown aetiology that usually involves the periarticular tissues of large joints. We report a rare case in which it presented in both index fingers in a 5-year-old boy. The tumours were resected and he made an uncomplicated recovery except for mild tapering deformity on the left. There has been no sign of recurrence a year later. PMID- 11878183 TI - Malignant chondroid syringoma of the hand: a case report. AB - We present a rare case of malignant chondroid syringoma of the hand in a 37-year old man. Ray amputation with coverage by an island reverse flow forearm flap gave a good final result. There have been no signs of recurrence during a follow up of five years. PMID- 11878185 TI - Soft cannabinoid analogues as potential anti-glaucoma agents. AB - Cannabinoids are able to reduce elevated intraocular pressure; however, their use in glaucoma treatment is not approved due to severe systemic side effects. New cannabinoid derivatives have been designed based on a retrometabolic/soft drug approach; they were expected to have local effect, but not systemic side effects. Lead compounds and soft analogues were prepared using Pechmann condensation. In agreement with the SAR hypothesis used for the present soft drug design, all the compounds that were successfully synthesized had IOP lowering effect, but the common metabolite of soft analogues that was found to be inactive. Accordingly, when the soft analogue 8 was administered i.v., its biological effect lasted just for 15 minutes; nevertheless, when administered topically, its effect lasted significantly longer. Its metabolite, though, was inactive when applied either i.v. or topically. Thus, the designed soft analogues proved to be good candidates for topical control of glaucoma without producing systemic side effects. The preliminary i.v. experimental data could be successfully described by an indirect response PK/PD model. PMID- 11878184 TI - Factors affecting the expression and function of P-glycoprotein in rats: drug treatments and diseased states. AB - The expression and function of P-glycoprotein (P-gp), an ATP-dependent efflux pump, were examined in rats pretreated with dexamethasone (DEX), an inducer of P gp, and in rats with glycerol-induced acute renal failure (ARF) and with CCl4 induced acute hepatic failure (AHF). DEX pretreatment increased the P-gp level and its functional activity in the intestine. In contrast, in ARF and AHF rats, the in vivo P-gp function was systemically suppressed, even though the level of P gp remained unchanged or rather increased. In Caco-2 cells, the plasma collected from diseased rats exhibited a greater inhibitory effect on P-gp function than did plasma from control rats. A higher-plasma level of corticosterone, an endogenous P-gp substrate/inhibitor, was observed in the disease rats. These findings indicate that the actual in vivo function of P-gp cannot be predicted merely from the expression level of P-gp, and suggest that some endogenous P-gp related compounds such as corticosterone participate in the regulation of in vivo P-gp function in diseased states. PMID- 11878186 TI - Design, pharmacokinetic, and pharmacodynamic evaluation of soft anticholinergics based on tropyl alpha-phenylcyclopentylacetate. AB - Four new soft anticholinergic agents based on tropyl alpha phenylcyclopentylacetate, 15a, 15b, 18a, and 18b, were designed and synthesized. Receptor binding studies on the cloned human muscarinic receptors indicated that the new soft anticholinergic agents possessed moderate potency as pKi ranged from 6.7 to 7.6. Mydriatic studies in rabbit eyes revealed that the duration of the action of the new soft anticholinergics (8.5-11.0 h) were shorter than that of atropine (about 24 h) under pharmacodynamic equivalent dose, and one of them, 18a, showed even shorter than that of tropicamide. In addition, after unilateral administration, significant dilation of pupil in the control eyes was observed with tropicamide and atropine but not with soft drugs, suggesting the systemic activity of soft drugs was minimal. With their soft nature, the new soft anticholinergics displayed much shorter protective effect against carbachol induced bradycardia (about 30 min) than atropine (at least 60 min) in rats. In vitro and in vivo pharmacokinetic studies demonstrated that the soft anticholinergics were rapidly hydrolyzed into the corresponding inactive metabolites once they were introduced into the systemic circulation. PMID- 11878187 TI - Interdependent chemical-electrochemical steps in retrometabolism-based drug and safer chemical design. AB - An extension of the retrometabolic based drug (chemical) design concept, specifically the soft drug approach, to the family of nitrone compounds is presented. Nitrones oppose oxidative challenges by virtue of their ability to very rapidly trap free radical species that are more stable and biochemically less harmful than the original molecular fragments. Moreover, the spin adducts may undergo further transformations including reaction with a second radical and decomposition (hydrolysis) to hydroxylamines and carbonyl compounds. Nitrones and their spin adducts may generate nitric oxide in vivo, which, like nitrones themselves, exerts a number of diverse activities in phylogenetically distant species as well as opposing effects in related biological systems. It was described as a major messenger in the cardiovascular, immune, and nervous systems, in which it plays regulatory, signaling, cytoprotective, and cytotoxic effects. Nitrones play an important role in the synthesis of drugs belonging to chemically and pharmacologically very different classes. A combined chemical electrochemical synthesis of nitrones has been elaborated. These compounds may be obtained from aldehydes or ketones and N-substituted hydroxylamines. These reactions were performed directly, in situ in the electrochemical cell, where phenylhydroxylamine obtained by electroreduction of nitrobenzene derivatives reacts with the carbonyl compound introduced in the cell. The kinetic and thermodynamic parameters of the processes were determined by analyzing the adequate polarographic curves. Differences between purely chemical and mixed chemical-electrochemical methods are discussed. Analysis of the experimental data permits optimization of the investigated process from a preparative point of view. Effects of structural factors were systematically evaluated. The proposed method may be useful for combinatorial chemistry as well. PMID- 11878188 TI - Quantitative structure/property relationship analysis on aqueous solubility using genetic algorithm-combined partial least squares method. AB - The present study was initiated to generate a model of predicting aqueous solubility of substances from their molecular structure. For 211 drugs or drug like compounds, their topological indices were calculated by Molconn-Z software. The optimal subset of the descriptors for the prediction of aqueous solubility was determined by genetic algorithm in combination with partial least squares (PLS) method. Thirty-four descriptors were selected by this method. Using 29 of the descriptors selected, of which the scaled PLS coefficient was significant, the cross-validated predictive q2 was 0.785 with 19 principal components that was the optimal and the standard error of prediction was 0.676. Thus, it is suggested that the model obtained would exhibit a good performance in predicting the aqueous solubility of compounds. PMID- 11878189 TI - Effect of 2-hydroxypropyl-beta-cyclodextrin on the solubility, stability, and pharmacological activity of the chemical delivery system of TRH analogs. AB - To improve the aqueous solubility and stability of the chemical delivery system (CDS) of the thyrotropin-releasing hormone (TRH) analogs, 2-hydroxypropyl-beta cyclodextrin (HPBCD) has been attempted. TRH analogs were [Leu2]-TRH, [Nva2]-TRH and [Nva2, Pip3]-TRH. Excess amount of CDS was added in various HPBCD in water solutions (0%-50%, pH 6.5). The mixture was saturated by ultra-sonication for 1 h at 15 degrees C and filtered. The concentration of CDS in the filtrate (solubility) was determined with UV detector, and subsequently the stability was investigated. By HPBCD complexation, the aqueous solubility and stability (half life) of CDS were significantly improved from undetectable levels to about 15 mg/ml and 30 h, respectively. In pH 6.5 and 7.4 HPBCD solution, the degradation of CDS was mainly via acid catalyzed water addition reaction, thus, e.g. [Leu2] TRH-CDS was more stable in pH 7.4 than in pH 6.5 aqueous solutions. After lyophilizing the saturated CDSs in 50% HPBCD complex solutions, the amount of CDS in the complex was determined as 26.22, 26.79, and 30.34 mg/g for [Leu2]-TRH, [Nva2]-TRH and [Nva2, Pip3]-TRH, respectively. The half-life of [Leu2]-TRH CDS/HPBCD solid complex at 25 degrees C, 4 degrees C and -15 degrees C was about 100 days, 440 days and no detectable change in three months, respectively. Argon protected condition did not improve the stability of lyophilized [Leu2]-TRH CDS/HPBCD complex. Dimethyl sulfoxide although increased the solubility of [Leu2] TRH-CDS in the 50% HPBCD solution by 1.3 times, significantly decreased its stability by 6.6 times. After intravenous administration of CDS (in 30% HPBCD) at a dose of 10 mumole/kg in mice, compared to the vehicle control or the same dose of [Leu2]-TRH (in 30% HPBCD), a significant increase in pharmacological effect (decrease in barbiturate-induced sleeping time) was observed. These results demonstrate the usefulness of cyclodextrin in the formulation of the CDSs of TRH analogs. PMID- 11878190 TI - Targeted drug delivery to the central nervous system via phosphonate derivatives (anionic delivery system for testosterone). AB - An anionic chemical delivery system (aCDS) has been developed and applied to deliver testosterone (T) to the central nervous system (CNS). The delivery of a target compound is achieved through the use of a specific targetor moiety which is an (acyloxy)alkyl-phosphonate-type functional group. The T-aCDS readily penetrates biological membranes by passive transport due to its increased lipophilicity and enters the target organ. Hydrolytic cleavage by esterases provides a negatively charged, hydrophilic intermediate phosphonate compound (TP ), which is "locked in" the CNS and should provide sustained, site-specific release of the drug. In vitro and in vivo investigations in rats showed that methyl-pivaloyloxymethyl-17-testosterylphosphonate (T-aCDS) might function as an anionic chemical delivery system of testosterone. The concentration of T-aCDS decreased fairly rapidly in vitro. The half-lives (t1/2) in different organs are as follows: blood 4.48 min (r = 0.9388), lung 5.53 min (r = 0.9661), liver 2.82 min (r = 0.9498), and brain 7.37 min (r = 0.9972). Simultaneously with the disappearance of T-aCDS, testosterone-phosphonate (TP-) appeared as a main metabolite in increasing concentration. In vivo evaluations (tail vein 11.3 mg/kg in DMSO) found maximum T-aCDS brain levels 5-10 min after administration; they fell under the borderline of detectability (< 0.1 microgram/g) after 60 min. Maximum concentration of the decomposition product (TP-) was obtained at 30 min after administration; it did not decrease significantly during the study. Even if the phosphonate derivative of the secondary, hindered hydroxyl group in this product was fairly resistant to phosphorolytic attack, the design principle can work for other compounds. PMID- 11878191 TI - Studies on a soft glycopyrrolate analog, SG-1. AB - A short-acting soft drug analog (SG-1) of glycopyrrolate (G) was developed by retrometabolic design in order to minimize systemic side effects and optimize the therapeutic index. SG-1,3-(1'-hydroxycyclopentyl)phenylacetoxy-1-methyl-1- methoxycarbonylpyrrolidinium bromide, was synthesized by: (a) esterification of phenylacetic acid with N-methyl-3-pyrrolidinol by DCC to obtain N-methyl-3 pyrrolidinyl phenylacetate; (b) reaction of lithium salt of above phenylacetates with cyclopentanone to obtain N-methyl-3-pyrrolidinyl 3-(1' hydroxycyclopentyl)phenylacetate; and (c) quarternization with methyl bromoacetate in acetonitrile to give the designed product. To evaluate the pharmacological effect of SG-1, its mydriatic activity in rabbit eyes was compared to that of glycopyrrolate. At the pharmacodynamically equivalent doses (the lowest dose that induces the maximum response) of SG-1 (1%) and glycopyrrolate (0.1%), the mydriatic activities lasted for 5 and 100 h, respectively. Compared to glycopyrrolate, the intrinsic pupil dilation potency of SG-1 was lower (approximately 1/10th) but the duration of action was much shorter (< 1/20th) as SG-1 is susceptible to facile enzymatic hydrolysis/deactivation in the rabbit eyes. In vitro metabolism and stability investigations further supported this finding. In vitro half lives of SG-1 in rat plasma, blood, and 20% liver and lung tissue homogenates were 15.62, 53.86, 263.43, and 318.35 min, respectively. In human plasma and blood, half-lives were 19.93 and 88.32 min, respectively. SG-1 was relatively stable under acidic conditions (pH 5 and lower). SG-1 is a promising, clinically useful short acting anticholinergic. PMID- 11878192 TI - 28-day oral toxicity study with soft corticosteroid BNP-166 in rats and dogs, followed by a 14-day recovery period. AB - The aim of the present study was to evaluate the soft corticosteroid BNP-166 in rats and dogs treated orally with 0.2, 2.0, and 20.0 mg/kg for 28 days and the reversibility of any abnormalities during a 14-day post-dosing period. The test substance, BNP-166, was well tolerated during the 28-day treatment period. The observed changes were all characteristic for the pharmacological actions of a glucocorticoid. Treatment related changes occurred in the adrenals and thymus, and, to a lesser extent, in the lymph nodes, spleen and liver. There were no statistically significant reductions in the cortisol levels of all groups in the 0.2 and 2 mg/kg treatments. Significant reductions were observed in the high-dose group (20 mg/kg), but levels returned to normal by the end of the 14-day recovery period. Based on the results, the No Observable Adverse Effect Level (NOAEL) of BNP-166 soft corticosteroid in rat and dog after 28-day oral administration is 2 mg/kg. This value is approximately 40 times higher than that of budesonide. Pharmacodynamic and receptor binding studies have shown BNP-166 to have a similar potency to budesonide; therefore, BNP-166 can be considered safer when administered orally than other corticosteroids such as prednisolone or budesonide. PMID- 11878193 TI - Novel transdermal targeting in steroid therapy: evaluation of PK/PD profiles using an animal model of rheumatoid arthritis. PMID- 11878195 TI - Physicochemical aspects of the enzymatic hydrolysis of carboxylic esters. AB - Considering the important role played by enzymatic hydrolysis in the metabolism of therapeutic agents designed by retrometabolic approaches (soft drugs and chemical delivery systems), the present article offers a review of a number of issues related to the enzymatic hydrolysis of carboxylic esters. Current knowledge regarding interorgan- and interspecies variability, stereospecificity, activation energy, proposed mechanism, and quantitative structure-metabolism relationship is summarized. The effects of chain-length and branching in the alcohol or acyl substituent on the rate of hydrolysis in congener series are also summarized. Available in vitro human blood data suggest that shortest half-lives are achieved with sterically non-hindered chains that are neither too short nor too long and are of around four carbon-atom long. PMID- 11878194 TI - Drug therapy in asthma bronchiale in the new millennium. AB - Asthma bronchiale represents a major health issue in industrialized countries and will likely remain so for decades. The drug treatment of asthma demonstrates certain peculiarities: revolutionary new drug introductions happen almost each quarter century. With improved understanding of asthma pathogenesis and drug metabolism, the potential for specific targeted and constructed therapies has become evident. Monoclonal antibodies to IgE and certain cytokines such IL-4 and IL-5 are being investigated as possible treatments for asthma. Similarly, preliminary studies of selective phosphodiesterase inhibitors in asthmatic patients have been encouraging. Other potential therapies include for example inhibitors of cytokine synthesis, promoters of Th2-Th1 switch, adenosine receptor agonists or antagonists, etc.. A new way is represented by a modified retrometabolic drug design resulting in so-called soft drugs. The first representative of this new drug class is loteprednol etabote (LE), a non fluorinated glucocorticoid approved for the allergic ophthalmological indications and now in clinical trial for the treatment of allergic airway diseases. Today's intensive search for new treatments should ensure a greater diversity of therapeutic possibilities for the management of asthma in the new millennium. PMID- 11878196 TI - The use of chemically modified cyclodextrins in the development of formulations for chemical delivery systems. AB - Retrometabolic drug design provides a highly useful and directed approach for identifying new drug candidates with improved therapeutic indices based on predictable/controlled metabolism and/or site-targeted delivery. In the process, formulation becomes an important and integral concern especially for brain targeting chemical delivery systems (CDS) based on the need for appropriate dosage form stability, solubility and dissolution characteristics. Adjuncts that have been useful in this regard are chemically modified, water soluble cyclodextrin derivatives such a 2-hydroxypropyl-beta-cyclodextrin (HP beta CD). These starch-derived excipients can interact with drugs via dynamic complex formation resulting in a number of beneficial pharmaceutical effects including increased apparent water solubility and stability as well as improved aesthetic and excipient compatibility properties. This cyclodextrin is approved in a number of product in the US and world-wide. HP beta CD has contributed to the development and preclinical/clinical testing of a number of CDS including E2 (estradiol)-CDS, AZT (zidovudine)-CDS, DEX (dexamethasone)-CDS and a neuropeptide CDS based on an enkephalin derivative. In these contexts, HP beta CD provided for stable and water-soluble dosage forms intended for parenteral administration. PMID- 11878197 TI - Synthesis and antitumour evaluation of 4-bromophenyl semicarbazones. AB - 4-Bromophenyl semicarbazone derivatives have been synthesized and their chemical structures have been confirmed by means of their IR, 1H-NMR data and by elemental analyses. The in vitro evaluation in the 3-cell line, one dose primary anticancer assay is described. The 4-bromo substituted p-nitrobenzylidene phenyl semicarbazone (5) showed significant activity against breast MCF7 cell line and was further evaluated for potential anticancer activity in an in vitro human disease-oriented tumour cell line screening panel that consisted of 59 human tumour cell lines arranged in nine subpanels, representing diverse histologies. Melanoma UACC-62 cell line was relatively more sensitive to compounds 5 (growth inhibitions: GI50 = 15.3 mumol/l). PMID- 11878198 TI - Effects of sub-minimum inhibitory concentrations of gatifloxacin on the inhibition of Staphylococcus aureus and Escherichia coli adherence. AB - The aim of this study was to investigate the capacity of subinhibitory concentrations of the newly developed fluoroquinolone antibiotic gatifloxacin (CAS 160738-57-8) to interfere with the mechanism of bacterial adhesion. Human buccal epithelial cells were incubated with different strains of both Staphylococcus aureus and Escherichia coli, grown in the presence of subinhibitory gatifloxacin concentrations varying from 1/2 MIC (minimum inhibitory concentration) to 1/128 MIC. A significant decrease was observed in the adherence of S. aureus and E. coli to buccal cells at drug MICs of 1/2 to 1/32 and 1/2 to 1/64, respectively. A large number of filamentous forms of different lengths and shapes were observed in the case of E. coli, whereas there was an abnormal increase in the diameter of the cells in the case of S. aureus. The interpolation of these pharmacodynamic findings with the pharmacokinetic curve indicate that the effect of subinhibitory concentrations of gatifloxacin can prolong its antimicrobial effects against S. aureus and E. coli for as long as 30 h and 37 h, respectively, after the MIC values have been reached. PMID- 11878199 TI - Anti-Mycobacterium tuberculosis agents derived from quinoxaline-2-carbonitrile and quinoxaline-2-carbonitrile 1,4-di-N-oxide. AB - In this paper new quinoxaline derivatives with different substituents in positions 3, 6, 7 and 8 are reported. Their biological activities against Mycobacterium tuberculosis have been assessed and most of the 1,4-di-N-oxide derivatives have been shown to strongly inhibit the bacteria growth in the first in vitro screening. One of these N-oxides (4) is a promising candidate due to its good Selectivity Index (7.95). On the other hand, those compounds without N-oxide moieties showed no or very low activity (growth inhibition: 17% and 39%). PMID- 11878200 TI - Synthetic derivatives of curcumin and their activity against Leishmania amazonensis. AB - In a previous work, the in vitro and in vivo activity of a series of diarylheptanoid derivatives against Leishmania amazonensis has been described. Based on the promising results, ten new compounds belonging to the same chemical class were synthesized and have been investigated in relation to their leishmanicidal activity. The compounds were obtained through several chemical modifications on the basic structure of curcumin (1,7-bis-(4-hydroxy-3 methoxyphenyl)-1,6-heptadiene-3,5-dione) in an attempt to increase its effectiveness and decrease the potential toxic effects. The drugs were assayed in vitro against L. amazonensis promastigotes and using pentamidine isethionate as reference drug. The results showed that the most effective compound is 1,7-bis-(4 propargyl-3-methoxyphenyl)-1,6-heptadiene-3,5-dione, which is about ten times more efficient than the original curcumin. Nevertheless, these results did not allow us to make any correlation between the leishmanicidal activity and the chemical structure of the compounds. PMID- 11878201 TI - From the manual method of Topliss to a modified quantitative method. AB - The optimization of the properties of a lead compound is the first goal of most pre-clinical research projects. Optimization strategies may be applied to the synthesis of analogous compounds in order to minimize cost and time. One strategy of synthesis is the change of the substituents in the molecule. The manual method of Topliss was introduced for the prediction of the substituted compounds that will have the most potent activity in a series of aromatic substituted analogues. A modified Topliss method is proposed that consist of the quantitative correlation by a single regression equation of the activity of a series of 4 or 5 substituted aromatic compounds with the descriptor parameters: hydrophobic (pi), electronic (sigma) and sterics (Es and MR) and some combinations of them in order to predict future synthesis, or to obtain a training set of compounds to be used in the application of more advanced experimental design methods. These results when compared with those of multiple regression analysis applying the Hansch equation are very satisfactory. PMID- 11878202 TI - [Comparison beta-blockers and their relevance to pharmacologic therapy of cardiovascular diseases]. AB - beta-Blockers have regained interest in the treatment of cardiovascular diseases after a period of decreasing importance due to the introduction of other cardiovascular treatment principles. beta-blockers expose the longest list of indications in cardiovascular therapy available and present positive mortality results for the treatment of arterial hypertension, the postinfarction situation and heart failure. There are three indications for the treatment of atrial fibrillation alone: slowing of AV-conductance, sinus rhythm restoration and atrial stabilization. Among the most recent developments there is the clear establishment of evidence for their life prolonging activity in heart failure treatment which has been proven for carvedilol, bisoprolol and metoprolol in NYHA (New York Heart Association) stages 2 and 3 and so far for carvedilol only in NYHA stage 4. Differences between beta-blocking agents are clinically most relevant for the presence of an intrinsic sympathomimetic activity (ISA) which should no longer be present in clinically used compounds. The ISA of bucindolol is the most likely reason for its failure in an outcome study for heart failure treatment. Other pharmacological distinctions relate to the selectivity in the beta-adrenergic system, activity in the alpha-adrenergic system and differences in metabolism and elimination. PMID- 11878203 TI - General pharmacological profile of the novel muscarinic receptor agonist SNI 2011, a drug for xerostomia in Sjogren's syndrome. 2nd communication: effects on somatic nervous system and on autonomic nervous system and smooth muscle. AB - A novel muscarinic receptor agonist SNI-2011 ((+/-)-cis-2-methylspirol[1,3 oxathiolane-5,3'-quinuclidine] monohydrochloride hemihydrate, cevimeline, CAS 153504-70-2), is a candidate therapeutic drug for xerostomia in Sjogren's syndrome. The general pharmacological properties of this drug on the somatic nervous system and on the autonomic nervous system and smooth muscle were investigated in mice, rats, guinea pigs, rabbits and cats. 1. Somatic nervous system: SNI-2011 had no effect on the neuromuscular junction in rats and no muscle relaxant effect in mice. No surface anesthetic effect was observed in guinea pigs, but infiltration anesthetic effect was found after intracutaneous injection of solution (1% or higher). 2. Autonomic nervous system and smooth muscle: SNI-2011 tended to cause mydriasis at 3 mg/kg i.v. or higher in rabbits and dose-dependently caused mydriasis at 10 mg/kg p.o. or higher in rats. Mydriasis in rats was also observed by ophthalmic instillation, caused via the peripheral muscarinic acetylcholine receptors. SNI-2011 elevated the base line tension of nictitating membrane in cats when it was injected intravenously at 3 mg/kg or higher. In the smooth muscle, SNI-2011 increased the spontaneous movement of isolated rabbit ileum (1 x 10(-6) mol/l or higher), contractions of isolated guinea pig ileum (1 x 10(-6) mol/l or higher) and isolated guinea pig trachea (3 x 10(-6) mol/l or higher). SNI-2011 relaxed the histamine- and noradrenaline-induced contractions of isolated guinea pig aorta and augmented noradrenaline- and phenylephrine-induced contractions of isolated rat vas deferens. These effects were induced by relatively higher concentrations only i.e. 1 x 10(-5) mol/l or higher. From these results, SNI-2011 has muscarinic side effects on the somatic nervous system and on the autonomic nervous system and smooth muscle, however, in the case of oral administration, that is clinical administration route, SNI-2011 caused no muscarinic side effect at the effective doses needed for saliva secretion. PMID- 11878204 TI - Quantification of procyanidins in oral herbal medicinal products containing extracts of Crataegus species. AB - According to the European Pharmacopeia a photometric assay is used for the estimation of procyanidins in Crataegi fructus. This assay is also most commonly used for procyanidin analysis in herbal medicinal products (HMPs) containing extracts of hawthorn (Crataegus species). In order to find an appropriate method for the determination of oligomeric and polymeric procyanidins by analysing various preparations containing extracts of Crataegus, the Ph. Eur.-method was compared to an HPLC-method with chemical reaction detection (HPLC-CRD-method) and another conventional photometric assay using 4-dimethylamino-cinnamic-aldehyde (DMACA). Total procyanidins estimates obtained with the pharmacopeial method were, depending on the reference standard used, at least more than 50% higher than those obtained with the DMACA-assay. The determination of individual procyanidins could only be achieved by HPLC-CRD. Monomeric, dimeric, and trimeric procyanidins could be separated and detected individually, whereas no HPLC separation was possible for higher polymeric compounds. However, these compounds could be analysed as co-eluting groups. Using the DMACA method for the estimation of total oligomeric procyanidins and the HPLC-CRD method for quantification of the mono- up to trimeric procyanidins, some market leading herbal medicinal products from Germany containing extracts Crataegus species (C. monogyna Jacq., C. laevigata D.C., C. pentagyna Waldst. et Kit., C. nigra Waldst. et Kit, C. azarolus L.) were analysed. Procyanidin B2 (epicatechin-(4 beta-->8)-epicatechin) was isolated from Aesculus hippocastanum fruit shells as reference standard for calibration purposes. The structure elucidation was carried out by by means of MS and 1H-NMR. Quantitative 1H-NMR spectroscopy (qNMR) was applied for purity assessment. PMID- 11878206 TI - Cancer incidence in Alaska Natives thirty-year report 1969-1998. AB - The Alaska Native Tumor Registry includes data from 1969 to the present. This report provides incidence rates over the thirty year period, 1969 through 1998, and compares trends over time for Alaska Natives (AN) with those of US Whites and Blacks. To examine current rates, average annual age-adjusted incidence rates for AN for 1984-98 are compared with US Whites. Data from the registry document numerous differences in rates of occurrence of specific cancers compared to US Whites and Blacks. Studies of these differences may provide clues to the causes and risk factors for the cancers. Most importantly, these data show that although cancer was considered a rare disease in the Alaska Native population as recently as the mid-twentieth century, the incidence rate for all cancers combined among Alaska Natives is now as high as that of US Whites, and even higher in women. On the other hand, despite relative differences in rates, the most frequently diagnosed cancers among Alaska Natives are the same as US Whites. Cancers of the lung, colon/rectum, breast, and prostate are most frequently diagnosed among Alaska Natives and in the U.S. These four cancers comprise over 50% of all diagnosed invasive cancers. Cancer of the lung is almost entirely preventable by eradication of tobacco use. Screening and early detection have been proven to reduce mortality for cancers of the colon/rectum and breast. Primary and secondary prevention of these cancers could markedly improve morbidity and mortality. PMID- 11878205 TI - Effects of antihistamines on leukotriene and cytokine release from dispersed nasal polyp cells. AB - In this study the effects of antihistamines on the release of eicosanoids and the pro-inflammatory cytokine tumor necrosis factor alpha (TNF alpha) were compared. Enzymatically dispersed cells from human nasal polyps served as an in vitro model of chronic respiratory mucosal inflammation. Nasal polyp cells (2 x 10(5)/ml) were sensitized with human IgE pre-incubated azelastine (CAS 58581-89-8), terfenadine (CAS 50679-08-8), levocabastine (CAS 79516-68-0) or cetirizine (CAS 83881-51-0), and stimulated with anti-human immunoglobulin E (IgE). Thromboxane B2 (TBX2) and leukotriene C4 (LTC4) were measured by radioimmunoassay (RIA), TNF alpha by enzyme-linked immunosorbent assay (ELISA). Data represent mean values of % inhibition estimated from the untreated positive control or mean IC50 (n = 5). Azelastine and terfenadine inhibited TNF alpha release with IC50 values of 6.2 mumol/l and 4.3 mumol/l, respectively. Terfenadine reduced TXB2 release by 37 +/- 15%, and LTC4 release was decreased by azelastine and terfenadine very potently by 86% and 100%, respectively. Azelastine shows anti-inflammatory properties in therapeutically relevant concentrations as assessed by its ability to reduce TNF alpha release as well as its ability to inhibit LTC4 production in allergically stimulated human nasal polyp cells. PMID- 11878207 TI - Health care's new consumers: boomers, bobos, health seekers, and the wired retired. PMID- 11878208 TI - "Bobos": health care's new trendsetters. PMID- 11878209 TI - The new consumer: marketing in the millennium. PMID- 11878210 TI - [Experience in surgical treatment of patients with focal forms of temporal epilepsy]. AB - Fifteen patients with focal lesions of temporal epilepsy were examined before and 6 months to 4 years after surgery. There were low-grade gliomas in 13 cases, muscle tissue heterotopy in 1 case, and medial temporal sclerosis in 1. Epileptogenic lesions were localized in the medial structures of the temporal lobe in 10 cases and in the lateral temporal regions in 5 cases. Comprehensive neurophysiological study, including intraoperative electrocorticography, was used to identify an epileptogenic zone (EZ). EZs were found in the medial structures on the affected side in all patients. Lesionectomies were performed in all cases. It was combined with medial resection in 12 cases (it was done in 10 patients until epileptic activity disappeared). Nine patients stopped having seizures after surgery. One patient had only auras. Improvements were slight in 5 patients. A relationship was found between the surgical outcomes and the site of a lesion and EZ. They authors conclude that complete recovery may occur in cases with medial locations of a lesion and EZ. The outcomes of surgery are much poorer in cases with the lateral sites of an lesion and in those with the medial site of EZ. PMID- 11878211 TI - [Hypophyseal adenoma in elderly persons. Experience with surgical treatment of 102 patients]. AB - In 1972 to 2000, a total of 102 patients aged 60 years or more underwent surgery for pituitary adenomas. They were 54 males and 48 females (mean age 63.5 years). Before surgery, all the patients had impaired vision (with asymmetry of visual loss in 61% of patients). 17% had ophthalmoplegia. Most adenomas (92%) were clinically and biochemically hormonally inactive. In all cases, pituitary adenomas had extrasellar extension: 63% of the adenomas were enclosed and 37% were invasive. A total of 111 interventions were made in 102 patients. Analyzing the outcomes indicates that transsphenoidal microsurgery is an effective and safe treatment in elderly patients with pituitary adenomas. PMID- 11878212 TI - [Motor responses in transcranial magnetic stimulation and somatosensory evoked potentials in patients with pain syndromes]. AB - The main goal was to evaluate the motor and somatosensory systems by recording evoked motor responses (EMRs) during transcranial magnetic stimulation (TMS) and somatosensory evoked potentials (SEPs) in patients with neurogenic pain syndromes before and after implantation of the systems for chronic antipain epidural stimulation. Fifteen patients with neurogenic pain syndromes and a control group of 15 apparently healthy examinees were examined. The patients were found to have a significant reduction in the motor thresholds of EMRs during TMS and an increase in the amplitude of EMRs after implantations of the systems. There were no significant changes in the amplitude-time characteristics of short SEPs as compared to the healthy examinees and after implantation of the systems. Analysis of the amplitude-time characteristics of long SEPs in these patients revealed a significant increase in the amplitude of the component P250 as compared to the normal values and its decrease after implantation of the systems. PMID- 11878213 TI - [Problems in brain protection]. AB - The main aim of intensive care in acute head injury and in the postoperative period is to prevent, detect, and revert a secondary neuronal injury. To maintain the optimum systemic and cerebrovascular functions can substantially promote this aim achievement. There are some new neuroprotective interventions that are currently under investigation. Although the major focus of recent cerebral protection researches has been on the development of receptor-specific drugs, this effort has resulted in better outcomes. At present, patients are well served by using more traditional techniques to prevent and treat cerebral ischemic events. Initial therapy should include interventions to improve cerebral perfusion and the oxygen-carrying capacity of blood. Once this is accomplished, measures should be made to monitor blood glucose concentrations and to treat fever. General anesthetic choice may be of great importance in monitoring intracranial pressure and seizure activity. However, if direct cerebral protection is desired, a barbiturate should be the anesthetic of choice. All these measures should increase the chance of patients to neurologically recover following hypoxia and ischemia. PMID- 11878214 TI - [Immunomorphological aspects of formation of an intracranial aneurysm]. AB - An immunohistochemical study of the arteries of the circle of Willis and the walls of ampullar aneurysms indicated the impaired expression of the matrix proteins collagens I, III, and V in the aneurysms. Examination of the proliferative activity of cell elements in different parts of the aneurysmal sac showed a significant activation of proliferative processes along the cervical parts of aneurysms and the presence of proliferating cells in the body and fundus of aneurysms different in sizes. The findings suggest that the magnitude of proliferative and synthetic processes plays a definite role in the formation of an aneurysmal sac and in the so-called growth of aneurysms. PMID- 11878215 TI - [Microsurgical treatment of compression of the rear wall of the spinal artery in intervertebral arthrosis]. AB - Vertebral artery stenosis caused by cervical spondylosis is generally caused by compression of the artery by osteophytes arising from an uncinate process. Compression caused by facet joint osteophytes is rarely reported. The paper shows that compression may also occur posteriorly by osteophytes from the facet complex. Careful evaluation of preoperative angiograms and computed tomographic or MRI scans is required to determine whether an anterolateral, posteriorlateral or posterior approach might be more beneficial for decompression. PMID- 11878216 TI - [Disseminated craniofacial chondrosarcoma]. AB - Chondrosarcomas are malignant mesenchymal tumors of chondroid nature. Less than 5% of all chondrosarcomas are localized in the head and neck. The paper analyzes a case of extensive chondrosarcoma of the skill base, which involves the anterior and median cranial fossa, ethmoidal, sphenoidal, right maxillary sinuses, and intratemporal fossa. A basal approach that is a combination of two-flapped subfrontal and orbitozygomatous accesses was used to remove the tumor. In doing so, the tumor could be excised and extensive defects of the skull base could be effectively closed with the flaps of the periostium and musculus temporalis, yielding a good functional and cosmetic effect. PMID- 11878217 TI - [Periocular skin depigmentation, developing after removal of a cranioorbital meningioma]. AB - The paper gives a case of periocular skin depigmentation developing after neurosurgical treatment for hyperostotic cranioorbital meningioma. It also shows a natural history of the tumor, as seen in a series of sequential CT scans. The patient was operated on 13 years after the onset of the disease. Five months following surgery, a well-defined periocular depigmentation appeared on the operated side. This remained unchanged on a follow-up visit at months 12. As follows from the presented review of known cases of periocular depigmentation, the described postneurosurgical depigmentation has not yet been reported. PMID- 11878218 TI - [External liquorrhea as a result of a spinal gunshot wound]. PMID- 11878219 TI - [Neurocytoma of the brain]. PMID- 11878220 TI - [Discoveries, inventions, and patents of the Professor A. L. Polenov Russian Institute of Neurology (on the 75th anniversary of its foundation)]. PMID- 11878221 TI - [Plastic surgical repair of the base of the skull after removing a craniofacial tumor]. PMID- 11878222 TI - Validation of a heteroscedastic hazards regression model. AB - A Cox-type regression model accommodating heteroscedasticity, with a power factor of the baseline cumulative hazard, is investigated for analyzing data with crossing hazards behavior. Since the approach of partial likelihood cannot eliminate the baseline hazard, an overidentified estimating equation (OEE) approach is introduced in the estimation procedure. It by-product, a model checking statistic, is presented to test for the overall adequacy of the heteroscedastic model. Further, under the heteroscedastic model setting, we propose two statistics to test the proportional hazards assumption. Implementation of this model is illustrated in a data analysis of a cancer clinical trial. PMID- 11878223 TI - Covariate order tests for covariate effect. AB - A new approach for constructing tests for association between a random right censored life time variable and a covariate is proposed. The basic idea is to first arrange the observations in increasing order of the covariate and then base the test on a certain point process defined by the observation times. Tests constructed by this approach are robust against outliers in the covariate values or misspecification of the covariate scale since they only use the ordering of the covariate. Of particular interest is a test based on the Anderson-Darling statistic. This test has good power properties both against monotonic and nonmonotonic dependencies between the covariate and the life time variable. PMID- 11878224 TI - Optimal weight functions for marginal proportional hazards analysis of clustered failure time data. AB - The choice of weights in estimating equations for multivariate survival data is considered. Specifically, we consider families of weight functions which are constant on fixed time intervals, including the special case of time-constant weights. For a fixed set of time intervals, the optimal weights are identified as the solution to a system of linear equations. The optimal weights are computed for several scenarios. It is found that for the scenarios examined, the gains in efficiency using the optimal weights are quite small relative to simpler approaches except under extreme dependence, and that a simple estimator of an exchangeable approximation to the weights also performs well. PMID- 11878225 TI - Estimating distributions with increasing failure rate in an imperfect repair model. AB - A failed system is repaired minimally if after failure, it is restored to the working condition of an identical system of the same age. We extend the nonparametric maximum likelihood estimator (MLE) of a system's lifetime distribution function to test units that are known to have an increasing failure rate. Such items comprise a significant portion of working components in industry. The order-restricted MLE is shown to be consistent. Similar results hold for the Brown-Proschan imperfect repair model, which dictates that a failed component is repaired perfectly with some unknown probability, and is otherwise repaired minimally. The estimators derived are motivated and illustrated by failure data in the nuclear industry. Failure times for groups of emergency diesel generators and motor-driven pumps are analyzed using the order-restricted methods. The order-restricted estimators are consistent and show distinct differences from the ordinary MLEs. Simulation results suggest significant improvement in reliability estimation is available in many cases when component failure data exhibit the IFR property. PMID- 11878226 TI - Inferences concerning exponential distributions in the presence of randomly right censored data with missing censored values. AB - Inferences concerning exponential distributions are considered from a sampling theory viewpoint when the data are randomly right censored and the censored values are missing. Both one-sample and m-sample (m > or = 2) problems are considered. Likelihood functions are obtained for situations in which the censoring mechanism is informative which leads to natural and intuitively appealing estimators of the unknown proportions of censored observations. For testing hypotheses about the unknown parameters, three well-known test statistics, namely, likelihood ratio test, score test, and Wald-type test are considered. PMID- 11878227 TI - [Portrait of N.I. Pirogov, done by a contemporary]. PMID- 11878229 TI - [Changes in the hemomicrocirculatory system in various forms of intestinal blockage]. PMID- 11878228 TI - [Theory of spatial organization of epithelial layers (using neuroepithelia as an example)]. AB - New principles of spatial organization of epithelial layers and highly productive approach for studying their tridimensional histoarchitecture have been proposed for the first time. This approach was based on conception of module structure of tissues and represents a family of topological and geometrical models of tissue structure and their experimental aprobation. This approach allows to create theoretical histology of epithelial layers, allowing to predict and define experimentally new variants of multi-row and multi-serie epithelia and to forecast their changes in development and pathology. According to this conception a family of new tridimensional tissue models was created. It was demonstrated that the structures studied are characteristics for real epithelial. Possibility of existence of a new type epithelia was predicted and a complex of new informative signs for their spatial organization was presented. PMID- 11878230 TI - [Changes in the ultrastructure of neuroglia in some regions of the rat brain in manganese chloride poisoning]. AB - Ultrastructure of astrocytes, olygodendrocytes and microgliocytes of caudate nucleus, substantia nigra and frontoparietal area of the cortex were studied under the influence of the everyday peroral treatment with MnCl2.4H2O during 30 days. Experimental animals were divided into two groups. Animals from the first group received 20 mg/kg of MnCl2.4H2O while for animals from the second one the dose made 50 mg/kg. Animals were killed on days 30 and 120 of the experiment. It was demonstrated that chloride manganese intoxication influences the ultrastructure of all types neuroglial cells, the majority of both macro- and microgliocytes remaining in normal state. Cytotoxic effect of manganese ions on astrocytes manifests in mitochondria destruction, widening of canaliculi and endoplasmic net cisterns, glycogenic granules accumulation, which leads to reduction of oxidative processes level in the nervous tissue. Increased phagocytic activity of the astrocytes was displayed in conditions of relatively high doses of MnCl2.4H2O. Microgliocyte reaction was distinctly dose-dependent and was manifested through their phagocytary activity increase. Oligodendrocytes were most stable to chloride manganese influence. PMID- 11878231 TI - [Neuronal organization of the cortical amygdalic nucleus in the rat brain]. AB - Cytoarchitectonic peculiarities and neuronal organization of anterior (COa) and posterior (COp) cortical nuclei of the rat brain were studied on five levels:rostral and caudal levels of anterior and posterior regions appropriately and transitional hippocampal level. Neuronal composition characteristic for every level and interrelations between neurons of certain layers were demonstrated. The authors opinion is that COa and COp should be classified as interstitial structures, occupying the position between nuclear and screen centres of the nervous system. All the structures of the cortical nucleus portion described are characterized by original neuronal organization. PMID- 11878232 TI - [Morphofunctional characteristics of gastrin-releasing peptide synthesizing system of the hypothalamus in normal conditions and in experimental diabetes in rats]. AB - 5 weeks development of streptozoticin-induced diabetes mellitus in the rats is accompanied with the increase of gastrin-releasing peptide (GRP) immunopositive neurons amount in parvocellular neurosecretory subdivisions of immunopositive fibers increased in these structures:the content of GRP increased in median eminence as well. In supraoptic nucleus and posterior magnocellular subdivision of paraventricular nucleus the amount of GRP-immunopositive neurons increased by the second week of diabetes development with its decrease by the fifth week. Thus, the increase of GRP synthesis in hypothalamic neurosecretory structures in diabetes mellitus may be considered as a compensatory reaction directed on the activation of the central mechanisms of feeding restriction and stimulation of insulin synthesis in the pancreas. PMID- 11878233 TI - [Subpial nerve plexus in the dog spinal cord]. AB - Spinal cord of 10 mature dogs and 5 pups were studied using silver nitrate impregnation and staining after Nissl, Van-Gieson and hematoxylin-eosin. Subpial neuronal layer representing a plexus of non-myelinated and single myelinated nerve fibres and also single neurons and their accumulations was distinguished throughout the whole spinal cord length. Relatively even segmentary distribution of neurons in the subpial plexus was observed in pups. In mature animals neurons of subpial neuronal layer were accumulated predominantly at the level of cervical thickening (3-6 neurons) and less in lumbar thickening (2-4 neurons). In newborns and adult animals maximal neuronal number was found in the subpial layer located on dorsolateral surface of the spinal cord substantiaa alba. PMID- 11878234 TI - [Structural organization of the acellular region of the sinoauricular nerve plexus in the dog heart at maximal motor activity]. AB - The reactions of the dog sinoatrial node artery under hypo- and hyperkinesia were revealed by the serial semi-thin section morphometric analysis. Single maximal physical load may be a cause of significant changes in the main source of blood supply of the dog heart sinoauricular region. During 4 weeks period hypokinezia leads to decrease of structural and functional reserve of the artery. The following physical load causes acute reduction of the blood volume in the region of the right atrium of the heart. The individual peculiarities of the artery adaptive reactions are determined by the level of the heart functional activity and by the cardiac muscle metabolism, which should be taken into account when using maximal physical loads in clinic and in experimental studies. PMID- 11878235 TI - [Anatomic features of the coeliac trunk]. AB - The study was performed in 120 specimens using preparation. According to the experimental data, from two to six branches originated from coeliac trunk. Division into three branches was the most often variant (in 60.8%). Division into two arteries was revealed in 5.0% cases. Only in one of 120 specimen six branches branching from coeliac trunk were found. In one of the specimen studied combined branching of coeliac trunk with superior mesenterial artery from coeliac aorta was found. PMID- 11878236 TI - [Accidental involution of the thymus after splenectomy (an experimental study)]. AB - Using immunological, general clinical and luminescent histochemical methods experimental study of immunological status and bioamine-containing thymus structures was performed on third, seventh and fifteenth postsplenectomy days in 90 outbred albino male rats. Splenectomy leads to significant changes in blood immunological parameters and morphofunctional state of thymus bioamine-containing structures. Significant increase of biogenic amines content in all thymus structures, leukocyte number, complement activity neutrophilic granulocytes phagocytic activity, were registered in 3 days. Further growth of bioamine content, decrease of cortical and medullary substances diameter, significant number growth of juvenile forms of mast cells were observed in 7 days. Essential decline in erythrocyte number, hemoglobin and IgG and IgA content, leukocytosis with leukemoid reaction development were defined in blood formula. On 15th postsplenectomy day maximal biogenic amine concentration, especially histamine was reached and further reduction in cortex and medulla as well as decline of all the immunoglobulins and decrease of neutrophilic granulocytes phagocytosis were determined. Thus, splenectomy and subsequent development of accidental involution leads to secondary immunodeficiency and severe anemia. PMID- 11878237 TI - [Vasopressin and inflammation]. AB - At least three vasopressinergic brain systems are involved in inflammatory reaction. The first one is VP neurons of the bed nucleus of stria terminalis, projecting to antipyretic center in the preoptic forebrain region. The second system is neurons of the parvocellular subnucleus of the hypothalmic paraventricular nucleus. VP from these neurons reaches portal blood through the external zone of the median eminence. VP potentiates the effect of corticotropin releasing hormone on ACTH cells in the anterior pituitary. Synergetic modulation of pituitary-adrenal axis by these neurohormones maintains synthesis and release of glucocorticosteroids. The latter plays crucial role as endogenous immunosuppresants. The third VP system is magnocellular hypothalamic neurons that release vasopressin into general circulation. Systemic VP is capable of compensating the water lost caused by symptoms of inflammation. The review is devoted to analysis of current information concerning physiology and interactions between these systems during acute and chronic inflammation. The special reference has been made on the magnocellular neurons and the role of circulating vasopressin in the renal function regulation. PMID- 11878238 TI - [Morphofunctional characteristics of cell death in the epidermis in gunshot damage to rat skin under experimental conditions]. AB - Different types epidermocyte death were considered in a model of skin gunshot injury in albino rats, Using light microscopy, cytophotometry and immunocytochemistry. Epidermocyte death was characterized by various morphological changes of nuclei, spatial localization of dying cells and different intensity of cellular death, that was registered in all the experimental terms. Cell alterations of necrotic, oncotic and apoptotic types were observed. Oncotic and necrotic type keratinocyte death prevailed in early and middle stages of regenerative histogenesis due to structural and metabolic organic disorders in the skin: in later stages distinctly manifested apoptotic cell death is more possible. However it is more probable that the majority of such cells completes their cycle with necrosis. This is connected to the fact that apoptosis is an energy-dependent process as caspase-activating adapter proteins contact with mitochondrial membranes. Gunshot injury had likely caused definite changes in molecular mechanisms of proliferative processes regulation, cell differentiation and death, leading to accumulation of pathologically altered cells in the epidermis. PMID- 11878239 TI - [Morphofunctional changes in the small intestinal mucosa in dynamics of physiological pregnancy and lactation]. AB - Histostructural changes of small intestine mucosa of intact, pregnant and lactating rats were studied by optic and electron microscopy and morphometry. Increase of mucosa thickness, vili height, crypt depth, quantity of enterocytes and crypts were studied in dynamics of pregnancy. Period of lactation that followed labour was characterized by opposite processes, i.e. decrease of above mentioned parameters, which became identical to that in control animals. PMID- 11878240 TI - [Ultrastructural changes of epitheliocytes and vessels of the small intestinal microcirculatory system in rabbit pups infected with cholera vibrions]. AB - 10-12 days old suckling rabbits-pups were infected by cholera vibrions of 01329 serogroup (16063 and M045 strains). Ultrastructural changes in small intestinal epitheliocytes and microcirculatory vessels were studied. Both strains caused typical cholerogenic syndrome, dystrophic and necrotic changes in epitheliocytes, epitheliocytes and lymphocytes, increase of vessel permeability, rheological disorders. Focal cytoplasmic were formed, widening of intercellular spaces were observed, although the zone of dense contact was not disturbed. M045 strains caused destruction of dense contacts, resulting in loose arrangement of epitheliocytes and their division from the basement membrane. PMID- 11878241 TI - [Post-traumatic repair of the tibia in rats]. AB - Posttraumatic bone repair was studied histologically in 88 adult male Wistar rats. Thick, semithin and thin sections were stained with standard methods and investigated (microanatomy, histology, EM). The experimental animals were divided into five main groups: 1) control; 2) trained (swimming); 3) immobilized; 4) formalized (0.2 mg/kg i.m. every day); 5) alcoholised (3% alcohol instead of drinking water). The first group was divided into animals wit periost, contacted with endost and bone marrow and animals without periost contacts (with isolated periosteum). Bone perforation causes local hemorrhage and tissue destruction. Thr first reparative changes on the first-seventh days (proliferating fibroblasts and capillary sprouts grow into the blood clot and injured area, degranulation of neutrophils and tissue basophils, appearance of activated macrophages and osteoclasts) occur in soft tissue. Intensive collagen synthesis in fibroblasts began. On the fourteenth day collagen synthesis in osteoblasts was increased (packed collagen fibrils in vacuolated cytoplasm). Posttraumatic osteohistogenesis on d 4-21 was generally completed on d 28-42 bone formation (morphogenesis) and permanent remodeling were continued. Periosteal osteogenesis and bone repair require an existence of the periosteum contact with endosteum and bone marrow (growth and differentiation of endosteal bone callus was inhibited and those of periosteal callus was arrested in groups with isolated periosteum). Similar results were achieved in i.p. heterotopic autografts of repaired tissue in diffusion chamber (with isolated periosteal osteogenetic cells appearing only by 21st day after bone injury). In trained rats bone repair was stimulated, while in immobilised and in injured animals it was inhibited and resulted in chronic inflammation of bone marrow and abscesses. PMID- 11878242 TI - [Comparative bilateral morphometric characteristics of human finger bones]. AB - The morphometric parameters of fingers of the hand in teenage girls were studied using the measurement complex developed by the author. The quantitative characteristics of distal phalangers, individual finger segments and the whole fingers as local anatomical structures belonging to hand, are presented. The analysis of these parameters was performed from the point of view of principles of bilateral and radial symmetry, structural and functional lateralization and evolutionary development of hand of the prehensile type. The results obtained may find application in designing the peripheral devices for telecommunication and computer techniques. PMID- 11878243 TI - [Forms and methods of optimizing the teaching process in the Department of Normal Anatomy of the Russian Military-Medical Academy]. PMID- 11878244 TI - Proceedings of the IX International Symposium on Motor Control. Varna, Bulgaria, October 8-12, 2000. PMID- 11878245 TI - Cancer chemoprevention of INSAM (ginseng). Proceedings of a symposium. April 20, 2001. PMID- 11878246 TI - Creation of a neonatal end-of-life palliative-care protocol. PMID- 11878248 TI - Conflict of interest revisited. PMID- 11878247 TI - Detection of patient-ventilator asynchrony during low tidal volume ventilation, using ventilator waveform graphics. PMID- 11878249 TI - Festschrift for Keizo Sugimachi. PMID- 11878251 TI - [Abstracts of the 6th French Language Congress of Pneumology. Nice, France, 26-29 January 2002]. PMID- 11878250 TI - Spare the plow, save the soil. PMID- 11878252 TI - Profiles of editorial board members: North and South America. PMID- 11878253 TI - Making important health information available: Internet sites for women. PMID- 11878254 TI - Profiles of editorial board members: North American and African connections. PMID- 11878255 TI - Calculating medication dosages. PMID- 11878257 TI - Intravenous epoprostenol: a new therapy for primary pulmonary hypertension. PMID- 11878256 TI - A comparison of hydrostatic leveling methods in invasive pressure monitoring. PMID- 11878258 TI - Ethanol, isopropanol, methanol, and ethylene glycol poisoning. AB - Alcohol intoxication, commonly encountered in emergency department and clinic settings, is by no means a benign condition. Ethanol ingested alone or in combination with other CNS depressants (eg, isopropanol, methanol, ethylene glycol, sedatives, opioids) can be fatal. Obtaining the patient's history and careful observation for clinical signs and symptoms, along with appropriate analysis of results of laboratory tests, are the key to determining and differentiating the agent ingested. It is critical that poisoning due to ethanol and/or other related alcohols should be recognized early in order to initiate appropriate treatments and prevent fatalities. Emergency department nurses may be the first persons to collect the essential data, and it is incumbent upon them to plan and initiate appropriate care. In continuing management for these patients, critical care nurses must understand the factors contributing to the observed signs and symptoms in order to initiate and monitor ongoing care and prevent serious complications. PMID- 11878259 TI - Managing trauma patients with abdominal compartment syndrome. AB - ACS is due to a rapid increase in intra-abdominal pressure. Although ACS may occur in both surgical and nonsurgical patients, patients who have abdominal or pelvic trauma and/or require massive fluid replacement are at increased risk. Critical care nurses are in a unique position to recognize early signs and symptoms of increased intra-abdominal pressure to ensure timely intervention. Aggressive hemodynamic, pulmonary, and operative management is essential for the optimal outcome of patients with ACS. Without definitive treatment, multisystem organ dysfunction and death ultimately ensue. PMID- 11878260 TI - Monitoring pulmonary artery pressures: just the facts. AB - Numerous research-based guidelines for PA pressure monitoring are available. Despite the availability of this large body of literature related to PA pressure monitoring, critical care nurses continue to demonstrate insufficient knowledge and ability to apply information related to the collection and interpretation of data obtained with a PA catheter. This article focuses on these problematic areas, and reinforces the need to continue to include the basic principles of PA pressure monitoring in education and training programs. PMID- 11878261 TI - Another nursing shortage wake-up call??? H e l - l - l - l - o - o - o - o!! PMID- 11878262 TI - Setting up a clinic for congestive heart failure patients. PMID- 11878263 TI - Visual tools for families: a picture is worth a thousand words. PMID- 11878264 TI - Mucin-like domain of enteropeptidase directs apical targeting in Madin-Darby canine kidney cells. AB - Enteropeptidase, a type II transmembrane protein of the enterocyte brush border, is sorted directly to the apical membrane of Madin-Darby canine kidney II cells. Apical targeting appears to be mediated by an N-terminal segment that contains a 27-amino acid residue O-glycosylated mucin-like domain consisting of two short mucin-like repeats, A and B. Targeting signals within these repeats were characterized by using green fluorescent protein (GFP) as a reporter. Constructs with a cleavable signal peptide and both repeats A and B were secreted apically. Similar constructs lacking mucin repeats were secreted randomly. Either repeat A or B was sufficient to direct apical targeting of GFP. O-linked oligosaccharides alone were not sufficient for targeting because fusion to a different O glycosylated motif did not alter the random secretion of GFP, and several constructs with mutations in either repeat A or B were O-glycosylated and secreted randomly. In addition, repeat B appears to contain an apical targeting signal that functions in the absence of glycosylation. Density gradient centrifugation indicated that, unlike several other apically targeted membrane and soluble proteins, apical sorting of mucin-GFP chimeric proteins does not appear to utilize lipid rafts. PMID- 11878265 TI - Rescue of hematopoietic stem cells following high-dose radiation injury using ex vivo culture on endothelial monolayers. AB - Military personnel are at significant risk for potentially lethal myeloablative injury secondary to nuclear accident, nuclear attack, or chemical weapons attack. In an attempt to develop culture conditions in which hematopoietic stem cells might be rescued from the effects of radiation, we irradiated (1,000 cGy split dose) 6-week-old C57BL/6 (Ly 5.1) and syngeneic C57BL/6 (Ly 5.2) mice and tested whether bone marrow mononuclear cells (MNCs) harvested postirradiation could be rescued via coculture in porcine microvascular endothelial cell (PMVEC) monolayers. We found that a subpopulation of bone marrow MNCs exposed to 1,000 cGy could be maintained and expanded over 10 days in a PMVEC culture (3.8-fold expansion), whereas liquid suspension culture did not maintain a significant number of hematopoietic cells postirradiation. Colony-forming assays demonstrated that murine MNCs exposed to 1,000 cGy did not give rise to granulocyte/macrophage colony-forming units (CFU-GM), erythroid burst-forming units (BFU-e), or CFU-Mix in 14-day cultures, whereas irradiated MNCs that were subsequently cultured in PMVEC-generated CFU-GM, BFU-e, and CFU-Mix at cloning efficiencies of 1.3%, 0.2%, and 0.2%, respectively. In survival studies, we found that 78% of mice that were irradiated and then transplanted with irradiated/PMVEC-expanded MNCs were alive at day 50, compared with 18% of irradiated control mice (p < 0.05). We also observed that mice transplanted with irradiated/PMVEC-expanded MNCs showed complete hematologic recovery. At 8 weeks post-transplant, we found evidence of Ly 5.1 donor cells in both the bone marrow and the spleen of the transplanted animals, but the levels of engraftment were low (range, 0-5.1%; mean, 1.9%). These results demonstrate that a subpopulation of bone marrow stem cells are capable of surviving the effects of high-dose radiation if these cells are placed in coculture with endothelial cell monolayers. PMID- 11878266 TI - Upper facial asymmetries resulting from unilateral coronal synostosis. Diagnosis and surgical reconstruction. PMID- 11878267 TI - Two cases of severe iron-deficiency anaemia due to inflammatory bowel disease in the dog. AB - Two dogs were referred for investigation of lethargy and anaemia. Preliminary examinations by the referring veterinary surgeons had revealed severe anaemia, which was poorly regenerative. In one case, the anaemia was microcytic and, in the other, it was hypochromic. These findings were suggestive of chronic blood loss anaemia. Neither dog had external signs of gastrointestinal disease, although one had vomited on a single occasion during the two weeks preceding referral. Although both dogs were producing grossly normal faeces, further investigations detected faecal occult blood. Multiple full thickness gastrointestinal biopsies were taken at laparotomy. These showed segmental eosinophilic enteritis of the duodenum and proximal jejunum in one case, and marked chronic lymphocytic gastritis in the other. Both cases illustrate the fact that severe systemic disease can result from gastrointestinal pathology in the absence of clinical signs of gastrointestinal disease. PMID- 11878268 TI - Surgical treatment of congenital lobar emphysema in a puppy. AB - Congenital lobar emphysema was found in a three-month-old male springer spaniel. Clinical signs included coughing, progressive dyspnoea and abnormal lung auscultation. Radiographic examination revealed overinflation and hyperlucency of the right middle lung lobe. Following lung lobectomy, clinical signs resolved and the animal had normal exercise tolerance. Previous case reports are reviewed and discussed in relation to human congenital lobar emphysema. PMID- 11878270 TI - Calorimetry as a tool for predicting bulk viscosity drift of polyamic acid ionic salt solutions. PMID- 11878271 TI - Crystal structure of zinc(II) complex of a pyridine containing self-assembling system. PMID- 11878269 TI - Epoprostenol-induced pulmonary vasodilatation in patients with pulmonary hypertension measured by electrical impedance tomography. AB - Electrical impedance tomography (EIT) has been proposed as a method to monitor dynamic changes in the pulmonary vascular bed. In this study we examined the validity of EIT in the measurement of pulmonary vasodilatation in eight patients with primary and secondary pulmonary hypertension when given the vasodilating agent epoprostenol (Flolan). Therefore, catheterization of the pulmonary artery was performed in the ICU and the cardiac output was measured by means of the Fick method. The pulmonary vascular resistance (PVR) and mean pulmonary arterial pressure (mPAP) were determined. Epoprostenol was given in increasing doses to test reversibility of pulmonary hypertension. The maximum test dose was 12 ng kg( 1) min(-1). During each step simultaneous EIT (DAS-01 P Portable Data Acquisition System, Sheffield, England) measurements were performed with the 16 electrodes equidistantly positioned in the third intercostal space. The maximal systolic impedance change, relative to end-diastole, deltaZperf, was chosen as a measure of pulmonary perfusion. The impedance change between baseline and highest tolerable epoprostenol concentration was compared with the change in PVR. The mean PVR (dyn s/cm5) decreased from 636 (+/-399) to 366 (+/-242); p < 0.01. DeltaZperf (in arbitrary units) for the whole patient group increased from 901 (+/-295) x 10(-3) to 1082 (+/-472) x 10(-3) (p<0.05). Only one patient showed a reduction in pulmonary artery pressure >20%, which is defined as significant vasodilatation. A strong relationship was found between the impedance changes and the change in PVR and mPAP in the patient with a significant vasodilatation on epoprostenol. From these results we conclude that EIT is a reliable method to measure blood volume changes due to pharmacologically induced vasodilatation in the pulmonary bed. PMID- 11878272 TI - Bone mineral density and body composition and influencing factors in children with rheumatic diseases treated with corticosteroids. AB - In rheumatic diseases the use of corticosteroids (CS), immobility, or the disease itself, may cause osteoporosis and growth retardation. We evaluated bone mineral density (BMD) by dual energy X-ray absorptiometry (DXA), growth and physical activity in 27 children with rheumatic disease, all treated with high dose CS for at least one year, in a cross-sectional design. BMD SDS was significantly lower than zero: -1.02 for total body and -1.49 for lumbar spine measurement. SDS for fat mass was higher than zero. In multiple regression analysis the Child Health Assessment Questionnaire score significantly correlated with BMD of the lumbar spine. There was no significant correlation with cumulative dose or duration of CS treatment. Height SDS decreased during treatment to -1.57 (p <0.001 compared to 0). In conclusion, BMD and body composition in children with rheumatic disease treated with CS are influenced by physical activity, as well as corticosteroid treatment and type of rheumatic disease. PMID- 11878273 TI - Trichloroacetic acid (TCA) and trifluoroacetic acid (TFA) mixture toxicity to the macrophytes Myriophyllum spicatum and Myriophyllum sibiricum in aquatic microcosms. AB - Trichloroacetic acid (TCA) and trifluoroacetic acid (TFA) have been detected together in environmental water samples throughout the world. TCA may enter into aquatic systems via rainout as the degradation product of chlorinated solvents, herbicide use, as a by-product of water disinfection and from emissions of spent bleach liquor of kraft pulp mills. Sources of TFA include degradation of hydrofluorocarbons (HFCs) refrigerants and pesticides. These substances are phytotoxic and widely distributed in aquatic environments. A study to assess the risk of a binary mixture of TCA and TFA to macrophytes in aquatic microcosms was conducted as part of a larger study on haloacetic acids. M. spicatum and M. sibiricum were exposed to 0.1, 1, 3 and 10 mg/l of both TCA and TFA (neutralized with sodium hydroxide) in replicate (n = 3) 12000 l aquatic microcosms for 49 days in an one-way analysis of variance design. Each microcosm was stocked with 14 individual apical shoots per species. The plants were sampled at regular intervals and assessed for the somatic endpoints of plant length, root growth, number of nodes and wet and dry mass and the biochemical endpoints of chlorophyll a, chlorophyll-b, carotenoid content and citric acid levels. Results indicate that there were statistically significant effects of the TCA/TFA mixture on certain pigment concentrations immediately after the start of exposure (2-7 days), but the plants showed no signs of stress thereafter. These data suggest that TCA/TFA mixtures at environmentally relevant concentrations do not pose a significant risk to these aquatic macrophytes. PMID- 11878274 TI - Rapid assessment of macro algal cover on intertidal sediments in a nutrified estuary. AB - Macroalgal blooms have been considered to be an indicator of eutrophication. A new and rapid method is described for the assessment of macroalgal cover in the intertidal zone of estuaries. Macroalgal cover in the intertidal of the nutrient enriched River Deben estuary was found to reach a maximum of 50% coverage, but this varied seasonally with the highest percentage cover during June and July. Macro-algae mats were particularly associated with areas of hard substrata providing suitable attachment points, rather than with the nutrient concentrations along the estuary. The occurrence of macroalgae may be more related to the substrate than to the nutrient status of the estuary. PMID- 11878276 TI - Bioprosthetic heart valve leaflet deformation monitored by double-pulse stereo photogrammetry. AB - A double-pulse stereo photogrammetry technique has been developed for the dynamic assessment of the leaflet deformation of bioprosthetic heart valves under simulated physiological conditions. By using a specially designed triggering technique, which takes the advantage of the field transfer mechanisms of the charge coupled device camera, two consecutive images separated by a time interval as short as 5 ms were captured. This made it possible to investigate the realistic leaflet deformation during the valve opening and closing processes which typically last 25-45 ms. This technique was applied to assess a newly developed pericardial valve leaflet in a physiological pulse flow loop. Quantitative leaflet deformations of the valve opening and closing were generated from sequences of digital images. The results can later be applied to finite element analysis of bioprosthetic heart valve leaflet stress and strain during a complete cardiac cycle. PMID- 11878275 TI - Expressing creatine kinase in transgenic tobacco--a first step towards introducing an energy buffering system in plants. AB - Creatine kinase a key enzyme in cellular energy homeostasis of vertebrates offers the promise of engineering plants with enhanced stress tolerance. In order to provide plants with such an energy buffering system, tobacco was transformed with a cDNA, encoding the cytosolic brain-type isoform of chicken creatine kinase (BB CK), the expression of which was under the control of the cauliflower mosaic virus 35S (CaMV 35S) promoter. Transgenic tobacco plants were selected and suspension cultures generated. Both transgenic plants and suspension cultures were shown to stably express enzymatically active BB-CK in vitro and in vivo, and in most cases for three successive generations (T0-T2). Exogenously supplied creatine was shown to enter the plant cells and resulted in only a slight reduction in root growth at concentrations up to 10 mM. Furthermore, the BB-CK expressing tobacco plants and cell suspension cultures were able to convert creatine into phosphocreatine. PMID- 11878277 TI - Dental management of severe dentinogenesis imperfecta in a mild form of osteogenesis imperfecta. AB - Dentinogenesis Imperfecta (DI), in which the teeth are discolored, translucent and brittle, can occur in isolation as a familial trait and as a component of the skeletal dysplasia Osteogenesis Imperfecta (OI). In a Cape Town family, 20 persons in 3 generations had mild OI, with the additional manifestation of severe DI. The family was assessed at the Dental Genetic Unit of the University of the Western Cape and appropriate dental treatment was provided. In this setting, a detailed treatment plan was devised for a severely affected woman. This plan proved to be efficient and cost effective, and the final outcome was pleasing to the patient. Dentinogenesis Imperfecta is not uncommon and may well be encountered in conventional dental practice. The necessary clinical expertise is within the scope of the skills of the general dentist. PMID- 11878279 TI - Oral-facial-digital syndrome, Type I: a case report. AB - Oral-facial-digital syndrome is a group of congenital anomalies, which affects the face, oral structures and digits. There are nine subtypes. OFDS type I, is x linked dominant trait mostly affecting females. Reports of OFDS type 1 in Asians are extremely rare. This paper shows a case of OFDS type 1, in a southern Chinese girl, who in addition to most of the classic features, had fusion of the mandibular canine and lateral incisor teeth. PMID- 11878278 TI - Bacterial, behavioral and environmental factors associated with early childhood caries. AB - The goals of this cross-sectional study were to characterize and compare demographic, behavioral, and environmental factors potentially associated with early childhood caries (ECC) and to assess salivary levels of mutans streptococci (MS) and lactobacilli (LB) in underserved, predominantly Hispanic children. One hundred forty-six children aged 3 to 55 months with a range of caries experience were identified and examined. ECC was primarily associated with the presence of MS and lack of access to dental care. Salivary MS levels among young children with ECC were higher than would be expected in a dentally healthy population, but lower than levels reported among older children at high risk for caries. After adjustment for age, children with log10 MS > or = 3.0 or log10 LB > or = 1.5 were about five times as likely (OR=4.9, 95% CI=2.0, 12.0) to have ECC than those with lower bacterial levels. This study demonstrated a significant association between relatively low cariogenic bacterial levels and dental caries in infants and toddlers. Antibiotic use, exposure to lead, and anemia were not significantly associated with the number of decayed and filled surfaces or decayed and filled teeth. ECC correlated significantly with child's age and lack of dental insurance of the children, as well as inversely with both family income and the educational level of the mother of the child. PMID- 11878280 TI - Capillary gas chromatography with mass spectrometric and atomic emission detection for characterization and monitoring chlordimeform degradation in honey. AB - Capillary gas chromatography-mass spectrometry and capillary gas chromatography atomic emission detection have been successfully used to identify and monitor the main degradation products of chlordimeform when this compound is initially present in honey. The analysis of laboratory-spiked honey samples over 28 weeks revealed the occurrence of two degradation products: 4-chloro-o-toluidine (I) and N-formyl-4-chloro-o-toluidine (III). During this period the concentration of chlordimeform decreased to 7.5% of its initial value; the concentration of compound I increased gradually whereas compound III was present in a larger proportion and reached a maximum around the 14th week. PMID- 11878281 TI - Migration of (hydroxy)carboxylic acids in coelectroosmotic capillary electrophoresis. Influence of the electrolyte composition. AB - This work focused on the way several electrolyte components could affect the electroosmotic flow and the capillary electrophoretic migration of aliphatic or aromatic (hydroxy)carboxylic acids. The effects exerted by the electroosmotic flow modifier, hexadecyltrimethylammonium bromide, the addition of metal salt to the electrolyte and the absorbance provider (chromophore) used for indirect detection were investigated. A retention of the organic acids was demonstrated. Its magnitude was shown to depend on the amount of cationic surfactant adsorbed onto the capillary walls. The addition of sodium nitrate led to a remobilization of all the acids except glycolic acid. Moreover, the presence of the chromophore was shown to influence mainly the migration of the glycolic acid. PMID- 11878282 TI - Olfactory function in HIV-positive subjects. AB - The aim of this study was to reinvestigate previous reports of chemosensory dysfunction in HIV-positive subjects. Odor thresholds, odor discrimination and odor identification were assessed using the Sniffin' Sticks test battery. Seventy four HIV-positive patients were tested. According to CDC criteria, 38 subjects were classified as stage A, 10 as stage B and 26 as stage C. None of the subjects exhibited severe cognitive impairment. Compared to normative data all subjects had normal odor identification and discrimination. However, odor thresholds were well below the median of a normal population. There were no significant differences between stage A, B or C subjects. This may be interpreted as indicating that olfactory dysfunction is among the primary deficits of HIV infection and occurs independently of disease stage. These results confirm previous work suggesting that odor thresholds are elevated early in HIV infection whereas a decline in identification and discrimination abilities is correlated with reduced cognitive abilities. PMID- 11878283 TI - Complementary and alternative medicine use among health plan members. A cross sectional survey. AB - CONTEXT. Many health plans have started to cover the cost of complementary and alternative medicine (CAM). National survey data indicate that CAM use is highly prevalent among adults. However, little is known about CAM use among health plan members. OBJECTIVE: To describe CAM users, the prevalence of CAM use, and how CAM use relates to utilization of conventional preventive services and health care satisfaction among health plan members. DESIGN: Cross-sectional mail survey in 1997. SETTING: Managed care organization in Minnesota. SAMPLE: Random sample of health plan members aged 40 and older stratified by number of chronic diseases; 4404 (86%) of the 5107 returned completed questionnaires. MEASURES: Use of CAM, patient characteristics (e.g., chronic diseases, health status), health behaviors (e.g., smoking, diet, exercise), and interaction with conventional health care (e.g., use of preventive services, having a primary care doctor, health care satisfaction). RESULTS: Overall, 42% reported the use of at least one CAM therapy; the most common were relaxation techniques (18%), massage (12%), herbal medicine (10%), and megavitamin therapy (9%). Perceived efficacy of CAM ranged from 76% (hypnosis) to 98% (energy healing). CAM users tended to be female, younger, better educated, and employed. Users of CAM reported more physical and emotional limitations, more pain, and more dysthymia but were not more likely to have a chronic condition. CAM users were slightly more likely to have a primary care provider (86% vs. 82% had chosen a primary care provider; P =0.014) and had more favorable health-related behaviors. CAM users and nonusers were equally likely to use conventional preventive services and were equally satisfied with their health plan. CONCLUSION: CAM use is highly prevalent among health plan members. CAM users report more physical and emotional limitations than do nonusers. CAM does not seem to be a substitute for conventional preventive health care. PMID- 11878284 TI - Information transfer in entrained cortical neurons. AB - Cortical interneurons connected by gap junctions can provide a synchronized inhibitory drive that can entrain pyramidal cells. This was studied in a single compartment Hodgkin-Huxley-type model neuron that was entrained by periodic inhibitory inputs with low jitter in the input spike times (i.e. high precision), and a variable but large number of presynaptic spikes on each cycle. During entrainment the Shannon entropy of the output spike times was reduced sharply compared with its value outside entrainment. Surprisingly, however, the information transfer as measured by the mutual information between the number of inhibitory inputs in a cycle and the phase lag of the subsequent output spike was significantly increased during entrainment. This increase was due to the reduced contribution of the internal correlations to the output variability. These theoretical predictions were supported by experimental recordings from the rat neocortex and hippocampus in vitro. PMID- 11878285 TI - A spatially continuous mean field theory of electrocortical activity. AB - A set of nonlinear continuum field equations is presented which describes the dynamics of neural activity in cortex. These take into account the most pertinent anatomical and physiological features found in cortex with all parameter values obtainable from independent experiment. Derivation of a white noise fluctuation spectrum from a linearized set of equations shows the presence of strong resonances that correspond to electroencephalographically observed 0.3-4 Hz (mammalian delta), 4-8 Hz (mammalian theta), 8-13 Hz (mammalian alpha) and >13 Hz (mammalian beta) activity. Numerical solutions of a full set of one-dimensional nonlinear equations include properties analogous to cortical evoked potentials, travelling waves at experimentally observed velocities, threshold type spike activity and limit cycle, chaotic and noise driven oscillations at the frequency of the mammalian alpha rhythm. All these types of behaviour are generated with parameters that are within ranges reported experimentally. The strong dependence of the phenomena observed on inhibitory-inhibitory interactions is demonstrated. These results suggest that the classically described alpha may be instantiated in a number of qualitatively distinct dynamical regimes, all of which depend on the integrity of inhibitory-inhibitory population interactions. PMID- 11878286 TI - A method using sustainability indicators to compare conventional and animal friendly egg production systems. AB - The objective of this paper was to describe a method that enables a comparison of egg production systems for their contribution to sustainable development of egg production, using data from the Netherlands for demonstration purposes. One conventional system, the battery cage system, and two animal-friendly systems, the deep-litter and the aviary systems, illustrate this method. The method is based on a three-phase framework that identifies relevant issues regarding sustainable development (Phase 1), translates issues into sustainability indicators (Phase 2), and assesses the contribution of sustainability indicators to sustainable development (Phase 3). Phases 1 and 2 are based on analysis of literature and consultation with experts. Phase 3 is based on graphical and numerical comparisons. A graphical comparison presents the relative deviation between actual and target values for each sustainability indicator. A numerical comparison combines the relative deviation for each sustainability indicator into an overall contribution of an egg production system to sustainable development. Sustainability indicators selected include economic performance, ammonia emission, energy use, hen welfare, farmer welfare, and egg quality. Based on equal importance of these indicators, the battery cage system shows the least negative contribution to sustainable development. The aviary system is considered a better animal-friendly alternative for a battery cage system when compared with a deep-litter system. The aviary system especially needs to improve economic performance and farmer welfare. The conclusion can only be tentative, however, because methodological aspects such as selection and weighting of sustainability indicators will need careful attention and further research. PMID- 11878287 TI - Extramammary Paget's disease found by abnormal vulvar brush sampling. AB - INTRODUCTION: Doctors are usually reluctant to perform a vulvar biopsy on a patient with non-specific chronic vulvitis--especially because of the rarity of vulvar malignancy in young women--until the lesion is suspected of being malignant. Therefore, most cases of extramammary Paget's disease (EMPD) were originally misdiagnosed as chronic and recurrent vulvar lesions. Late diagnosis of invasive lesions occurring in elderly females have resulted in cases of death. CASE: A 37-year-old patient showed an extended lesion on the vulva and perineum. In addition, abnormal cells were found from a vulvar scrape smear, and a following punch biopsy was used to diagnose and determine the extension of the disease. CONCLUSION: Diagnosis and demarcation of EMPD remain difficult due to the multifocal lesions and subtle nature of the disease. Brush sampling taken from suspicious areas can be a guide for multiple biopsies to demarcate the lesion before major surgery. A brush biopsy is presented as a first-step method to detect vulvar malignancy. PMID- 11878288 TI - Drug resistance in epithelial ovarian cancer: P-glycoprotein and glutation S transferase. Can they play an important role in detecting response to platinum based chemotherapy as a first-line therapy. AB - OBJECTIVE: Drug resistance is important for the treatment of ovarian cancer. P glycoprotein and glutation S-transferase as resistance markers play an important role in the effectivity of chemotherapeutical agents. The role of P-glycoprotein and glutation S-transferase in the treatment of epithelial ovarian cancer is not well understood. We investigated the relation between P-glycoprotein and glutation S-transferase level for response to platinum-based chemotherapy in epithelial ovarian cancer. MATERIAL AND METHODS: We reviewed 30 cases diagnosed as epithelial ovarian cancer and treated with platinum-based chemotherapy in the Department of Obstetrics and Gynecology, Akdeniz University School of Medicine. The material was attained from initial parafin-embeded blocks stained for P glycoprotein and glutation S-transferase. The cases that were diagnosed and treated before attending our clinic were not enrolled in the study. RESULTS: Mean age was 58.2 (25-70) and mean gravida 4.1 (0-10). Twenty-four patients (80%) were glutation S-transferase positive. Three cases (10%) out of 30 had positive reaction for P-glycoprotein. No difference was revealed regarding chemotherapy response rate among the cases showing glutation S-transferase positivity and P glycoprotein negativity. CONCLUSION: Detection of glutation S-transferase and P glycoprotein levels in epithelial ovarian cancer tissue is not important for response to platinum-based chemotherapy as a first line. PMID- 11878289 TI - Low frequency of enteric infections by Salmonella, Shigella, Yersinia and Campylobacter in patients with acute leukemia. AB - BACKGROUND: Several authors found that isolation of Salmonella, Shigella, Yersinia and Campylobacter spp. (SSYC) from stool cultures after the 3rd day of hospitalization is a rare event. The significance of enteric infections caused by these pathogens has not been systematically investigated in severely immunosuppressed patients with acute leukemia. PATIENTS AND METHODS: We screened all patients treated on the leukemia ward of a university medical center. A total of 1,185 stool cultures from 371 episodes of diarrhea, mostly following myelosuppressive chemotherapy, were examined for the complete range of classic bacterial enteric pathogens (i.e. SSYC). RESULTS: Only three (0.25%) cultures from one patient were positive for Salmonella enteritidis. This patient suffered from cholangitis. S. enteritidis could also be detected by liver biopsy. Other infections by classic enteric pathogens were not observed. CONCLUSION: Symptomatic infections by classic bacterial enteric pathogens in hospitalized patients with acute leukemia are very rare. Stool cultures for these pathogens cannot be recommended as a routine test in uncomplicated diarrhea occurring after the 3rd hospital day. PMID- 11878290 TI - Rapid diagnosis of tuberculous meningitis by ligase chain reaction amplification. AB - A total of 87 cerebrospinal fluid (CSF) samples obtained from 85 patients with suspected meningitis were examined for the presence of Mycobacterium tuberculosis by means of ligase chain reaction amplification (LCx; Abbott Laboratories). The results were compared with direct smear and culture results. Of 61 patients with pathological CSF, 9 (14.8%) were scheduled to receive treatment for tuberculous meningitis. The sensitivity of the smear and culture tests was 11.1 and 33.3%, respectively. The sensitivity, specificity, positive predictive value and negative predictive value of the LCx assay were 55.5, 100, 100 and 92.9%, respectively. The results reveal that amplification by ligase chain reaction is valuable for the diagnosis of tuberculous meningitis. PMID- 11878291 TI - Molecular epidemiological study of Salmonella enterica serovar paratyphi B infections imported from Turkey to Western Norway. AB - During the summer of 1999, several Norwegian tourists returning from Turkey became ill as a result of Salmonella enterica serovar paratyphi B (S. paratyphi B) infection. We examined the S. paratyphi B isolates from 14 of these patients (10 from blood cultures, 4 from stool specimens) who were admitted to 2 hospitals in Bergen, Norway during August and September 1999. Moreover, during the same period, a laboratory technician working at 1 of these hospitals was admitted with S. paratyphi B septicemia and was included in the study. Using repetitive sequence-based PCR (rep-PCR) with 2 primer pairs (ERIC and REP), pulsed-field gel electrophoresis and phage typing we found that the laboratory technician was infected with the same S. paratyphi B clone as the 14 tourists. The discriminatory capacity of the rep-PCR method and pulsed-field gel electrophoresis was examined using S. paratyphi B strains from the outbreak and from other geographical locations. We conclude that a combination of rep-PCR with the ERIC primer pair and phage typing was useful in discriminating between the epidemic isolates and epidemiologically unrelated isolates from S. paratyphi B infections and that the laboratory technician was most likely infected while handling patient samples or bacterial cultures from the Turkish tourists. PMID- 11878292 TI - Cutaneous nerve to the subacromial region originating from the lateral pectoral nerve. AB - During dissection practice, a cutaneous branch to the deltoid region, which originated from the lateral pectoral nerve, was found bilaterally in one Japanese male (two of 125 sides, 1.6%). The branch originated from the superior surface of the lateral pectoral nerve, ran on the superior surfaces of the coracoid process and the coraco-acromial ligament, and pierced the deltoid muscle close to the tip of the acromion. The distribution area of this cutaneous branch was similar to the cutaneous branch of the suprascapular nerve. Although the branch from the suprascapular nerve has been reported in man and primates, a minute description of such a branch from the lateral pectoral nerve is not currently available in the literature. According to the detailed analyses of the roots of the lateral pectoral nerve and the suprascapular nerve, the roots of both nerves are close to each other in the upper part of the superior trunk of the brachial plexus. Therefore, these cutaneous branches have different courses, but are considered to be a single nerve to complement the supraclavicular nerves. PMID- 11878293 TI - Impaired dental cytodifferentiation in glial cell-line derived growth factor (GDNF) deficient mice. AB - Glial cell line-derived neurotrophic factor promotes the survival of multiple neuron types in the central and peripheral nervous system. Moreover, it plays a key role in the development of the enteric nervous system and in the kidney organogenesis. Glial cell line-derived neurotrophic factor and their receptors are expressed in the developing tooth as well as in the trigeminal ganglion. However, the precise role of this growth factor in tooth morphogenesis and cell differentiation, or in the development of trigeminal ganglion cells, is still elusive. Using structural and ultrastructural techniques we analyzed in detail the first molar tooth germ of glial cell line-derived neurotrophic factor deficient mice as well as the neuronal density in trigeminal ganglion. The length and width of first molar tooth germ in knockout deficient animals showed no differences in the knockout animals in comparison with age-matched heterozygous or wild-type littermates. Nevertheless, in mice lacking glial cell line-derived neurotrophic factor, both ameloblasts and odontoblasts failed to fully develop and differentiate, and the enamel matrix and predentin layers were absent. On the other hand, the number of trigeminal sensory neurons and the structure of the nerves supplying first molar tooth germ were largely normal. Present results suggest a new non-neuronal role for glial cell line-derived neurotrophic factor in tooth development. Glial cell line-derived neurotrophic factor seems not to be involved in tooth initiation and morphogenesis, whereas it seems essential for cytodifferentiation. Conversely, neither development of trigeminal neuron nor nerve fibers supplying teeth are directly dependent on glial cell line-derived neutrophic factor. PMID- 11878294 TI - Image of the month. Leiomysarcoma. PMID- 11878295 TI - Omeprazole and ultrastructural modifications occurring in reflux esophagitis. PMID- 11878296 TI - Azathioprine intolerance in patients with IBD may be imidazole-related and is independent of TPMT activity. PMID- 11878297 TI - On finding flat adenomas: is the search worth the gain? PMID- 11878298 TI - A wider view on diagnostic criteria of nonalcoholic steatohepatitis. PMID- 11878299 TI - Prevalence and distinctive biologic features of flat colorectal adenomas in a North American population. PMID- 11878300 TI - Preliminary results of using a commercial fibrin sealant in the treatment of fistula-in-ano. AB - A prospective non-randomised study fibrin sealant injection to manage patients with fistula-in-ano, with magnetic resonance imaging (MRI) monitoring, was performed during the period 5/6/1999 to 28/2/2000. The aim was to determine whether a fibrin sealant could be used as a treatment modality for anorectal fistula and the usefulness of MRI perineum to monitor the disease activity. Ten patients were included in the study. Mean age was 47 years (range 7 months to 70 years). Male: female ratio was 9:1. Mean follow-up duration was 26.4 weeks. The overall success rate was 60%. The success rate of different fistula types were different (60%, 0%, 100% for intersphincteric, transphincteric, subcutaneous, respectively). Variable decrease in signal on STIR images and contrast enhancement was noted in the patients with successful and failure of fibrin sealant injection. In conclusion, fibrin sealant injection is a useful alternative treatment in the management of fistula-in-ano. MRI is helpful in delineating the anatomy of fistula-in-ano but not a useful tool to follow-up disease activity. PMID- 11878301 TI - Biliary surgery in sickle cell disease: the Jamaican experience. AB - Of 62 patients with homozygous sickle cell disease subjected to open cholecystectomy for symptomatic gallstones over a 12-year period at the University Hospital of the West Indies, 25 were males and 37 were females. Of these, 27 were paediatric patients aged 18 years or less, and 35 were adults. Preoperative transfusion was selectively administered. All cases presented with right upper quadrant pain and 15 of 62 with obstructive jaundice. Mucocoele of the gallbladder, empyema of the gallbladder and common bile duct stones were detected in 2, 2, and 23 patients, respectively. Exploration of the common bile duct was necessary in 31 cases and a T-tube sited in 15 cases. Twelve of the 62 patients developed acute chest syndrome post operatively (20%). There were 2 deaths, both occurring in patients who had developed acute chest syndrome; in a 34 year old and a 10 year old patient. Common bile duct related morbidity was proportionately more common in paediatric patients than adults, represented by ductal dilation (48% vs 37%), ductal calculi (44% vs 31%) and retained stones (7% vs 3%). PMID- 11878302 TI - Painless limp as a presentation of pelvic eosinophilic granuloma in a child. AB - Painless limp in children continues to pose a diagnostic challenge. In the majority of the cases transient synovitis of the hip is the cause. However, bone tumours should be considered as a possibility and specifically looked for when a child continues to have ongoing symptoms. Failure to diagnose tumours at an early stage can lead to a poorer outcome. This case report describes a child with eosinophilic granuloma who presented with a progressive painless limp. It emphasises the need to look carefully at the pelvic bones when evaluating a hip radiograph. PMID- 11878303 TI - A novel method of shortening an aortic graft. AB - We describe a simple method of shortening the abdominal aortic graft during an emergency procedure which is easy to perform and does not unduly delay the completion of the operation. PMID- 11878304 TI - Exposure of KS483 cells to estrogen enhances osteogenesis and inhibits adipogenesis. AB - Osteoblasts and adipocytes arise from a common progenitor cell in bone marrow. Whether estrogen directly regulates the progenitor cells differentiating into osteoblasts or adipocytes remains unknown. Using a mouse clonal cell line KS483 cultured in charcoal-stripped fetal bovine serum (FBS), we showed that 17beta estradiol (E2) stimulates the differentiation of progenitor cells into osteoblasts and concurrently inhibits adipocyte formation in an estrogen receptor (ER)-dependent way. E2 increased alkaline phosphate (ALP) activity and nodule formation and stimulated messenger RNA (mRNA) expression of core-binding factor alpha-1 (Cbfa1), parathyroid hormone/parathyroid hormone-related protein receptors (PTH/PTHrP-Rs), and osteocalcin. In contrast, E2 decreased adipocyte numbers and down-regulated mRNA expression of peroxisome proliferator-activated receptor-gamma (PPARgamma)2, adipocyte protein 2 (aP2), and lipoprotein lipase (LPL). Furthermore, the reciprocal control of osteoblast and adipocyte differentiation by E2 was observed also in the presence of the adipogenic mixture of isobutylmethylxanthine, dexamethasone, and insulin. Immunohistochemical staining showed that ERalpha and ERbeta were present in osteoblasts and adipocytes. A new mouse splice variant ERbeta2 was identified, which differed in two amino acid residues from the rat isoform. E2 down-regulated mRNA expression of ERalpha, ERbeta1, and ERbeta2. The effects of E2 are not restricted to the KS483 cell line because similar results were obtained in mouse bone marrow cell cultures. Our results indicate that estrogen, in addition to stimulation of osteogenesis, inhibits adipogenesis, which might explain the clinical observations that estrogen-deficiency leads to an increase in adipocytes. PMID- 11878305 TI - Incidence and natural history of Paget's disease of bone in England and Wales. AB - This study used a large, primary care, record-linkage resource (the General Practice Research Database [GPRD]) to evaluate the incidence, clinical presentation, and natural history of Paget's disease of bone in England and Wales. Between 1988 and 1999, we identified 2465 patients with the recorded diagnosis of Paget's disease of bone, within the five million subjects > or = 18 years old who were registered in the GPRD. The validity of diagnostic recording was assessed by questionnaire to individual general practitioners (GPs) in 150 patients; the diagnosis was confirmed in 93.8% of responders. The mean age of patients with Paget's disease was 75 years and 51% were men. The prevalence of the disorder was 0.3% among men and women aged > or = 55 years; incidence rates for clinically diagnosed Paget's disease rose steeply with age (men, 5 per 10,000 person-years; women, 3 per 10,000 person-years at the age of 75 years). Over the 11-year period of the study, the age- and sex-adjusted incidence rate of clinically diagnosed Paget's disease declined from 1.1 per 10,000 person-years to 0.7 per 10,000 person-years. Each patient with Paget's disease was matched to three controls matched by age, gender, and general practice. Cases had a greater risk of back pain (relative risk [RR], 2.1; 95% CI, 1.9-2.3), osteoarthritis (OA; RR, 1.7; 95% CI, 1.5-1.9), hip arthroplasty (RR, 3.1; 95% CI, 2.4-4.1), knee arthroplasty (RR, 1.6; 95% CI, 1.0-2.6), fracture (RR, 1.2; 95% CI, 1.0-1.5), and hearing loss (RR, 1.6; 95% CI, 1.3-1.9). Seven patients with Paget's disease developed a malignant bone neoplasm (0.3%). Using life table methodology, the estimated number of people who died within 5 years of follow-up was 32.7% among the patients with Paget's disease and 28.0% among the control patients. PMID- 11878306 TI - Midwifery education roundtable discussion: transitions to the workplace. AB - A snapshot of four graduates' views on their educational experience at a research institution as well as their perspective on how it prepared them for the challenges they faced in entering the workplace is presented. Discussants stated that research knowledge is a critical aspect to clinical practice. Their educational experience provided them with the ability to understand information technology applications, identify and access relevant scientific research, evaluate the integrity and comparability of research findings, and apply research findings to clinical practice. Areas within the curriculum that were identified as needing more content and/or greater emphasis included primary health care, how to work competently and effectively with persons from diverse cultural, socioeconomic, and racial and ethnic backgrounds, experience with public health providers, and content such as intimate partner violence and adolescent behaviors. PMID- 11878307 TI - Non-conventional yeasts. AB - In the beginning there was yeast, and it raised bread, brewed beer, and made wine. After many not days but centuries and even millenia later, it was named Saccharomyces cerevisiae. After more years and centuries there was another yeast, and it was named Schizosaccharomyces pombe; now there were two stars in the yeast heaven. In only a few more years there were other yeasts, and then more, and more, and more. The era of the non-conventional yeasts had begun. PMID- 11878308 TI - Evaluation of pKD1-based plasmid systems for heterologous protein production in Kluyveromyces lactis. AB - The stability of pKD1-based vectors was evaluated during the synthesis of intracellular and extracellular gene products in the yeast Kluyveromyces lactis. The Escherichia coli lacZ and MFalpha1 leader-BPTI (bovine pancreatic trypsin inhibitor) cassettes were placed under the control of the inducible K. lactis LAC4 promoter and inserted into the pKD1-based plasmids. To induce gene expression while maintaining inducer level, a gratuitous gal1-209 K. lactis strain was employed. Selective medium containing 5 g glucose/l and 0.5 g galactose (inducer)/l allowed optimum expression and secretion of heterologous products without a significant effect on the growth of the recombinant cells. During long-term sequential batch cultures (60 generations), plasmid instability was mainly the result of structural instability. The expression and secretion of BPTI resulted in greater structural instability relative to the intracellular beta-galactosidase. For both products, vectors carrying the pKD1 replication origin and the cis-acting stability locus (partial-pKD1 vectors) were more stable than vectors carrying the full pKD1 sequence (full-pKD1 vectors). However, after 55 generations, the beta-galactosidase and BPTI activities were still higher with the full-pKD1 vectors. This was due to the significantly higher initial beta galactosidase and BPTI activities for the full-pKD1 vectors (approximately 85% and 47% higher, respectively) relative to the partial-pKDI vectors. Southern blots confirmed that these increases were due to the higher copy number of the vectors carrying the full pKD1 sequence. In contrast to our previously reported results for the secretion of invertase, full-pKD1 vectors were preferred for the expression/secretion of beta-galactosidase and BPTI for at least 55 generations. Due to their structural stability, partial-pKD1 vectors will be advantageous for very long cultivation times. PMID- 11878309 TI - Cloning, characterization and comparison of the Pseudomonas mendocina polyhydroxyalkanoate synthases Phac1 and PhaC2. AB - This study describes a comparison of the polyhydroxyalkanoate (PHA) synthases PhaC1 and PhaC2 of Pseudomonas mendocina. The P mendocina pha gene locus, encoding two PHA synthase genes [phaC1Pm and phaC2pm flanking a PHA depolymerase gene (phaZ)], was cloned, and the nucleotide sequences of phaC1Pm (1,677 bp), phaZ (1,034 bp), and phaC2pm (1,680 bp) were determined. The amino acid sequences deduced from phaC1Pm and phaC2pm showed highest similarities to the corresponding PHA synthases from other pseudomonads sensu stricto. The two PHA synthase genes conferred PHA synthesis to the PHA-negative mutants P. putida GPp104 and Ralstonia eutropha PHB-4. In P. putida GPp 104, phaC1Pm and phaC2Pm mediated PHA synthesis of medium-chain-length hydroxyalkanoates (C6-C12) as often reported for other pseudomonads. In contrast, in R. eutropha PHB-4, either PHA synthase gene also led to the incorporation of 3-hydroxybutyrate (3HB) into PHA. Recombinant strains of R. eutropha PHB-4 harboring either P. mendocina phaC gene even accumulated a homopolyester of 3HB during cultivation with gluconate, with poly(3HB) amounting to more than 80% of the cell dry matter if phaC2 was expressed. Interestingly, recombinant cells harboring the phaC1 synthase gene accumulated higher amounts of PHA when cultivated with fatty acids as sole carbon source, whereas recombinant cells harboring PhaC2 synthase accumulated higher amounts when gluconate was used as carbon source in storage experiments in either host. Furthermore, isogenic phaC1 and phaC2 knock-out mutants of P. mendocina provided evidence that PhaC1 is the major enzyme for PHA synthesis in P. mendocina, whereas PhaC2 contributes to the accumulation of PHA in this bacterium to only a minor extent, and then only when cultivated on gluconate. PMID- 11878310 TI - Precision-cut organ slices as a tool to study toxicity and metabolism of xenobiotics with special reference to non-hepatic tissues. AB - Metabolism of xenobiotics is often seen as an exclusive function of the liver, but some current findings support the notion that the lungs, kidneys and intestine may contribute considerably. After the establishment of the use of liver slices as a useful in vitro model to study metabolism and toxicity of xenobiotics, the same concept is currently being used for slices from lung, kidney and intestine. It is the aim of this review to discuss the use of organ slices in biotransformation research. The basic idea behind the use of tissue slices in biomedical research is the assumption that the cells under study will function optimally in vitro if they are cultivated in an environment that is most alike to their natural in vivo embedding, which is the case in tissue slices. Advantages in the use of organ slices are the relatively easy preparation as well as the potential standardization of both the preparation and use. Moreover, a direct interspecies comparison can be made between liver, lungs, kidneys and intestines, for example with respect to their metabolic capacity and their sensitivity for toxicants. Of major importance is that organ slices can be made with a similar procedure from organs/tissues originating from different species, including man. This latter aspect is useful in drug development in general but also for a better insight in the metabolic fate of compounds in man. Importantly the use of slices may largely contribute to a reduction in the use of experimental animals. PMID- 11878311 TI - Rate-responsive pacing based on the atrio-ventricular conduction time. AB - Chronotropic incompetent patients are unable to adapt their heart rate adequately to the level of exertion. One treatment for this disease is a rate-responsive pacemaker that stimulates the heart at a rate proportional to an exertion-related variable derived from an implemented sensor (e.g., ventilation). The atrio ventricular conduction time (AVCT), which can act as such a sensor, corresponds to a well-defined interval in the intracardial electrogram. The AVCT is coupled to the sympathetic/parasympathetic activity of the autonomous nervous system (dromotropic effect), leading to a shortening of AVCT during exercise. We found that AVCT is also sensitive to the pacing frequency (pf). Hence, an AVCT-based pacemaker always constitutes a closed-loop system. General design rules for an AVCT pacemaker have been developed from our experimental results and a system theoretical treatment with reasonable assumptions. Problems addressed were uncertainties concerning the AVCT dynamics and disturbances in the signal. The following rules for the controller design have been derived: 1) The controller gain strongly depends on the therapeutic range of the pf, which is set by the medical doctor; 2) the closed-loop bandwidth of the system should be smaller than the respiratory frequency; and 3) a robust control strategy, capable of guaranteeing stability for a certain set of plant models, should be applied. PMID- 11878312 TI - Assessment of closed-loop ventilatory stability in obstructive sleep apnea. AB - Previous studies on ventilatory control in obstructive sleep apnea (OSA) have generally indicated depressed chemosensitivity, implying greater stability of the chemical control of breathing in these subjects. However, these results were based on tests involving steady-state or quasi-steady measurements obtained in wakefulness. We have developed a method for assessing the dynamic stability characteristics of chemoreflex control in OSA patients during sleep. While continuous positive airway pressure was applied to stabilize the upper airways, acoustically stimulated arousals were used to perturb the respiratory system during sleep. The fluctuations in esophageal pressure that ensued were analyzed, using a closed-loop minimal model, to estimate the chemoreflex loop impulse response (CLIR). Tests using simulated data confirmed the validity of our estimation algorithm. Application of the method to arousal responses measured in six OSA and five normal subjects revealed no statistically significant differences in gain margins and loop gain magnitudes between the two groups. However, the CLIR in the OSA subjects exhibited faster and more oscillatory dynamics. This result implies that, in addition to unstable upper airway mechanics, an underdamped chemoreflex control system may be another important factor that promotes the occurrence of periodic obstructive apneas during sleep. PMID- 11878314 TI - Abstracts of the 2002 Annual Meeting of the American Society for Clinical Pharmacology and Therapeutics. Atlanta, Georgia, USA. March 24-27, 2002. PMID- 11878313 TI - Adaptive filtering of evoked potentials with radial-basis-function neural network prefilter. AB - Evoked potentials (EPs) are time-varying signals typically buried in relatively large background noise. To extract the EP more effectively from noise, we had previously developed an approach using an adaptive signal enhancer (ASE) (Chen et al., 1995). ASE requires a proper reference input signal for its optimal performance. Ensemble- and moving window-averages were formerly used with good results. In this paper, we present a new method to provide even more effective reference inputs for the ASE. Specifically, a Gaussian radial basis function neural network (RBFNN) was used to preprocess raw EP signals before serving as the reference input. Since the RBFNN has built-in nonlinear activation functions that enable it to closely fit any function mapping, the output of RBFNN can effectively track the signal variations of EP. Results confirmed the superior performance of ASE with RBFNN over the previous method. PMID- 11878315 TI - A 16-year-old boy with back pain. AB - This case is presented to illustrate the imaging and clinical findings of a condition of interest to orthopaedic surgeons. The initial findings are noted on this page. The clinical and imaging diagnoses are presented on the following pages. PMID- 11878316 TI - A new cytotoxic fatty acid (5Z,9Z)-22-methyl-5,9-tetracosadienoic acid and the sterols from the far eastern sponge Geodinella robusta. AB - A new fatty acid, (5Z,9Z)-22-methyl-5,9-tetracosadienoic acid (1a), and a rare fatty acid, (5Z,9Z)-23-methyl-5,9-tetracosadienoic acid (2a), the predominant constituents of the free fatty acid fraction from the lipids of the sponge Geodinella robusta, were isolated and partly separated by reversed phase high performance liquid chromatography, followed by multifold crystallization from MeOH to give 1a and 2a in 70% and 60% purity, respectively. These fatty acids were identified as (5Z,9Z)-22- and (5Z,9Z)-23-methyl-5,9-tetracosadienoic acids by nuclear magnetic resonance techniques, including distortionless enhancement by polarization transfer, heteronuclear multiple quantum connectivity, and correlation spectroscopy experiments, as well as from mass-spectrometric data for their methyl esters, the methyl esters of their perhydro derivatives, and their pyrrolidides. Mixtures of 1a and 2a showed cytotoxic activity against mouse Ehrlich carcinoma cells and a hemolytic effect on mouse erythrocytes. The sterol fraction from the same sponge was analyzed by gas-liquid chromatography-mass spectrometry, and 24-methylenecholesterol was identified as a main constituent of this fraction. The implications of the co-occurrence of membranolytic long-chain fatty acids and 24-methylenecholesterol as a main membrane sterol are discussed in terms of the phenomenon of biochemical coordination. PMID- 11878317 TI - Fast and reproducible method for the direct quantitation of adipose tissue in newborn infants. AB - The role of body fat content and distribution in infants is becoming an area of increasing interest, especially as perception of its function appears to be rapidly evolving. Although a number of methods are available to estimate body fat content in adults, many are of limited use in infants, especially in the context of regional distribution and internal depots. In this study we developed and implemented a whole-body magnetic resonance imaging (MRI)-based protocol that allows fast and reproducible measurements of adipose tissue content in newborn infants, with an intra-observer variability of <2.4% and an inter-observed variability of <7%. The percentage total body fat for this cohort of infants ranged from 13.3-22.6% (mean and standard deviation: 16.6 +/- 2.9%), which agrees closely with published data. Subcutaneous fat accounted for just over 89% of the total body fat, whereas internal fat corresponded to almost 11%, most of which was nonabdominal fat. There were no gender differences in total or regional body fat content. These results show that whole-body MRI can be readily applied to the study of adipose tissue content and distribution in newborn infants. Furthermore, its noninvasive nature makes it an ideal method for longitudinal and interventional studies in newborn infants. PMID- 11878318 TI - Viral genome-linked protein (VPg) controls accumulation and phloem-loading of a potyvirus in inoculated potato leaves. AB - The viral protein covalently linked to the 5' end of the plus-sense, single stranded RNA genome of potyviruses (genus Potyvirus) can be an avirulence determinant in incompatible potyvirus-host combinations in which the resistance prevents systemic virus infection. The mechanism is not well known. This study shows that virus strain-specific resistance to systemic infection with Potato virus A (PVA) in Solanum commersonii is overcome by a single amino acid (aa) substitution, His118Tyr, in the viral genome-linked protein (VPg). Virus localization and other experiments revealed that Tyr118, controls phloem loading of PVA. The critical boundary may be constituted in phloem parenchyma, companion cells, or both. Tyr118 also controls the cellular level of virus accumulation in infected leaves, including phloem cells. Amino acid substitutions at three additional positions of the central part (aa 116) and C terminus (aa 185) of the VPg and of the N terminus of the 6K2 protein (aa 5) affect virus accumulation and rate of systemic infection but are not sufficient for phloem loading of PVA. These data, together with previous studies, indicate that the PVA VPg aa residues crucial for systemic infection are host specific. Also, our data and previous studies on other potyvirus-host species combinations indicate that the central part of the VPg is a domain with universal importance to virus-host interactions required for systemic invasion of plants with potyviruses. PMID- 11878319 TI - Sinorhizobium fredii HH103 has a truncated nolO gene due to a -1 frameshift mutation that is conserved among other geographically distant S. fredii strains. AB - Strain SVQ121 is a mutant derivative of Sinorhizobium fredii HH103 carrying a transposon Tn5-lacZ insertion into the nolO-coding region. Sequence analysis of the wild-type gene revealed that it is homologous to that of Rhizobium sp. NGR234, which is involved in the 3 (or 4)-O-carbamoylation of the nonreducing terminus of Nod factors. Downstream of nolO, as in Rhizobium sp. NGR234, the noeI gene responsible for methylation of the fucose moiety of Nod factors was found. SVQ121 Nod factors showed lower levels of methylation into the fucosyl residue than those of HH103-suggesting a polar effect of the transposon insertion into nolO over the noel gene. A noeI HH103 mutant was constructed. This mutant, SVQ503, produced Nod factors devoid of methyl groups, confirming that the S. fredii noeI gene is functional. Neither the nolO nor the noeI mutation affected the ability of HH103 to nodulate several host plants, but both mutations reduced competitiveness to nodulate soybean. The Nod factors produced by strain HH103, like those of other S. fredii isolates, lack carbamoyl residues. By using specific polymerase chain reaction primers, we sequenced the nolO gene of S. fredii strains USDA192, USDA193, USDA257, and 042B(s). All the analyzed strains showed the same -1 frameshift mutation that is present in the HH103 nolO-coding region. From these results, it is concluded that, regardless of their geographical origin, S. fredii strains carry the nolO-coding region but that it is truncated by the same base-pair deletion. PMID- 11878321 TI - Physiological Genomics of Cardiovascular Disease: From Technology to Physiology. Abstracts of the 2002 APS Conference. San Francisco, California, USA. February 20 23, 2002. PMID- 11878320 TI - Novel aspects of tomato root colonization and infection by Fusarium oxysporum f. sp. radicis-lycopersici revealed by confocal laser scanning microscopic analysis using the green fluorescent protein as a marker. AB - The fungus Fusarium oxysporum f. sp. radicis-lycopersici is the causal agent of tomato foot and root rot disease. The green fluorescent protein (GFP) was used to mark this fungus in order to visualize and analyze the colonization and infection processes in vivo. Transformation of F oxysporum f. sp. radicis-lycopersici was very efficient and gfp expression was stable for at least nine subcultures. Microscopic analysis of the transformants revealed homogeneity of the fluorescent signal, which was clearly visible in the hyphae as well as in the chlamydospores and conidia. To our knowledge, this is the first report in which this is shown. The transformation did not affect the pathogenicity. Using confocal laser scanning microscopy, colonization, infection, and disease development on tomato roots were visualized in detail and several new aspects of these processes were observed, such as (i) the complete colonization pattern of the tomato root system; (ii) the very first steps of contact between the fungus and the host, which takes place at the root hair zone by mingling and by the attachment of hyphae to the root hairs; (iii) the preferential colonization sites on the root surface, which are the grooves along the junctions of the epidermal cells; and (iv) the absence of specific infection sites, such as sites of emergence of secondary roots, root tips, or wounded tissue, and the absence of specific infection structures, such as appressoria. The results of this work prove that the use of GFP as a marker for F. oxysporum f. sp. radicis-lycopersici is a convenient, fast, and effective approach for studying plant-fungus interactions. PMID- 11878323 TI - Whom should the patient trust? A friendly report to Kevin Wildes. PMID- 11878322 TI - Regulation of rRNA gene number in Drosophila melanogaster: new aspects resulting from the use of free duplications. AB - We have isolated a bobbed (bb) mutant on the free duplication Dp(1;f)122bb(+) and we have measured the rDNA content of the bb(+) and the bb loci in genetic combinations in which none of the phenomena involved in the change of the rDNA redundancy occurs. We have also measured the rDNA content of the two bb loci carried by the free duplications in two different genetic combinations: (1) XXNO( )/Dp122bb(+) and XXNO(-)/Dp122bb females in which there are two attached X chromosomes completely deleted for the nucleolus organizer (NO) regions and therefore the only rDNA is contributed by the free duplication; (2) X/Dp122bb(+) and X/Dp122bb males, in which there are two bb loci, one on the X chromosome and the other on the X free duplication. The bb(+) and the bb duplications produced an overall increase of the rDNA content in the two genetic conditions tested. These results are not in favour of both a cis and trans effect of the regulator locus (cr(+) locus) hypothesised as being involved in the disproportionate replication of rRNA genes. PMID- 11878324 TI - Will charity laws close Catholic hospitals? PMID- 11878325 TI - Siamese twins--the surgery: an agonizing choice--parents, doctors, rabbis in dilemma. PMID- 11878326 TI - Siamese twins--the surgery: so one may live. PMID- 11878327 TI - Siamese twins: Rav Feinstein's ruling and the subsequent controversy. PMID- 11878328 TI - Conjoined twins: 2000 version. British Supreme Court's decision. PMID- 11878329 TI - Healthcare reform, rationing, and equity: a societal challenge. PMID- 11878330 TI - Halakhic guidelines for physicians in intensive care units. PMID- 11878332 TI - U.S. government views on the Declaration of Helsinki. PMID- 11878331 TI - Post-mortem sperm retrieval. PMID- 11878333 TI - Another meeting on the Helsinki Declaration. PMID- 11878334 TI - Difficulties in negotiating research access. PMID- 11878335 TI - Medical research: the role and responsibilities of doctors. AB - The General Medical Council in the UK has decided to join the already very large number of organisations, nationally and internationally, that issue guidelines on the conduct of research. This might be related to the fact that the UK, via the GMC, takes research misconduct more seriously than most countries, except that the GMC is developing separate guidelines in that area, in association with the Royal College of Physicians. So the purpose of yet another set of guidelines, albeit ones that will be professionally binding on doctors, is unclear. The draft guidelines are available for comment until 21 June and are reprinted below. PMID- 11878336 TI - Genetic research in Tonga. PMID- 11878337 TI - European Union clinical trials directive. PMID- 11878338 TI - Democracy and bioethical controversies. PMID- 11878339 TI - More research guidelines for comment. PMID- 11878340 TI - Ethics, home births and NHS trusts. AB - A long-time campaigner, for women to have a better and safer deal from the maternity services when giving birth, reviews the continuing difficulties many face when wanting to give birth at home. PMID- 11878341 TI - Genetic testing: policy issues for the new millennium. PMID- 11878342 TI - The challenge of validating genetic testing. AB - Validation of genetic testing is a multidisciplinary task that involves medical/clinical geneticists, other medical specialists, legislative authorities and the public. It is an area in which many issues are unresolved. Validation should not focus on the laboratory process alone, but should also include the prerequisites for service delivery, such as education and staffing, data collection and banking and information transfer, as well as pre- and post-test genetic counselling in a cost-effective perspective. It must thus be seen as an ongoing process at laboratory, familial and societal levels which involves many different actors. The challenges for the near future are to continue to develop technical standards for the laboratory and counselling procedures, and, in collaboration among professionals, lay persons and state authorities, to define a legal framework for the actions of the various players. This chapter discusses some problems related to the validation process for genetic testing, broadly defined. PMID- 11878343 TI - Genetic counselling: evolution or involution? AB - The need for genetic counselling derives from the peculiarities of genetic information, as compared to other biomedical tests, with particular reference to (a) its predictive character; (b) the existing gap between the ability to diagnose and to treat an inherited disorder, and (c) the psychological, social and ethical problems that genetic testing can raise. Counselling is traditionally performed by healthcare professionals, specifically trained to help individuals to develop ways of dealing with genetic information and gain a better understanding of the problems related with it. The growing number of genetic tests (for rare Mendelian as well as for common disorders), the development of easier and cheaper molecular techniques, the increasing tendency of physicians to have recourse to genetic tests, by-passing alternative diagnostic procedures, are all factors that contribute to the vast increase in the demand for genetic tests, a demand which is significantly out of step with the available numbers of trained counsellors. This paper discusses possible solutions, including the institution of committees with regulatory powers on genetic testing, the promotion of studies on models of genetic services, on programmes to monitor the services currently offered by test providers, and the expansion of training programmes and of employment opportunities for genetic counsellors. PMID- 11878345 TI - Population genetics and benefit sharing. AB - The majority of international or national guidelines, specific to human genetics concentrate on actual or potential clinical applications. In contrast, the Ethics Committee of the Human Genome Organisation (HUGO) attempts to provide guidance to the bench scientists engaged in fundamental research in genomics prior to any clinical applications. Often confused as constituting the Human Genome Project (HGP) itself, HUGO's (Human Genome Organization) ultimate goal is to assist in the worldwide collaboration underpinning the HGP. It is an international organisation with 1,229 members in approximately 60 countries. The Ethics Committee is one of HUGO's six international advisory committees. Composed of experts from a number of countries and disciplines, the HUGO Ethics Committee promotes discussion and understanding of social, legal, and ethical issues as they relate to the conduct of, and knowledge derived from, the Genome Initiative. Currently, it has 13 members from 11 difference countries. It has produced statements on the conduct of genetic research, on cloning, and, has most recently presented a 'Statement on Benefit-Sharing', April 11, 2000. The Intellectual Property Committee of HUGO has been active in the controversial area of patenting. The issue of benefit-sharing is one that has its source in the mandate of both committees. How to avoid both commodification of the person through payment for access to DNA and biopiracy with no return to benefits to the families or community? While patents are a legitimate form of recognition for innovation, there seems to be no therapeutic exception to some of its stringent rules and the 'morality' exclusion has lain dormant. The HUGO 'Statement on Benefit-Sharing' examines the issues of defining community, common heritage, distributive justice and solidarity before arriving at its conclusions in benefit sharing. This communication reviews some of these issues. PMID- 11878344 TI - Banking biological collections: data warehousing, data mining, and data dilemmas in genomics and global health policy. AB - While DNA databases may offer the opportunity to (1) assess population-based prevalence of specific genes and variants, (2) simplify the search for molecular markers, (3) improve targeted drug discovery and development for disease management, (4) refine strategies for disease prevention, and (5) provide the data necessary for evidence-based decision-making, serious scientific and social questions remain. Whether samples are identified, coded, or anonymous, biological banking raises profound ethical and legal issues pertaining to access, informed consent, privacy and confidentiality of genomic information, civil liberties, patenting, and proprietary rights. This paper provides an overview of key policy issues and questions pertaining to biological banking, with a focus on developments in specimen collection, transnational distribution, and public health and academic-industry research alliances. It highlights the challenges posed by the commercialization of genomics, and proposes the need for harmonization of biological banking policies. PMID- 11878347 TI - Setting up a hotline. AB - Consumer assistance hotlines represent one important model for delivering health care information to consumers. However, developing and managing such programs can be complicated. This issue brief presents some of the issues involved in setting up and maintaining a hotline. It also highlights the experiences of the Health Rights Hotline, an independent consumer assistance program in Sacramento, California. PMID- 11878346 TI - Chemical differentiation in regions of massive star formation. AB - We have reexamined the origin of the apparent differentiation between nitrogen bearing molecules and complex oxygen-bearing molecules that is observed in hot molecular cores associated with massive protostars. Observations show that methanol is an ubiquitous and abundant component of protostellar ices. Recent observations suggest that ammonia may constitute an appreciable fraction of the ices toward some sources. In contrast to previous theories that suggested that N/O differentiation was caused by an anticorrelation between methanol and ammonia in the precursor grain mantles, we show that the presence of ammonia in mantles and the core temperature are key quantities in determining N/O differentiation. Calculations are presented which show that when large amounts of ammonia are evaporated alkyl cation transfer reactions are suppressed and the abundances of complex O-bearing organic molecules greatly reduced. Cooler cores (100 K) eventually evolve to an oxygen-rich chemical state similar to that attained when no ammonia was injected, but on a timescale that is an order of magnitude longer (~10(5) yr). Hotter cores (300 K) never evolve an O-rich chemistry unless ammonia is almost absent from the mantles. In this latter case, a complex O-rich chemistry develops on a timescale of ~10(4) yr, as in previous models, but disappears in about 2 x 10(5) yr, after which time the core is rich in NH3, HCN, and other N-bearing molecules. There are thus two ways in which N-rich cores can occur. We briefly discuss the implications for the determination of hot-core ages and for explaining N/O differentiation in several well-studied sources. PMID- 11878348 TI - Social and health policy concerns raised by the introduction of the contraceptive Norplant. PMID- 11878349 TI - Peripherally hydrogenated neutral polycyclic aromatic hydrocarbons as carriers of the 3 micron interstellar infrared emission complex: results from single-photon infrared emission spectroscopy. AB - Infrared emission spectra of five gas-phase UV laser-excited polycyclic aromatic hydrocarbons (PAHs) containing aliphatic hydrogens are compared with the main 3.3 microns and associated interstellar unidentified infrared emission bands (UIRs). We show that neutral PAHs can account for the majority of the 3 microns emission complex while making little contribution to the other UIR bands; peripherally hydrogenated PAHs produce a better match to astrophysical data than do those containing methyl side groups; 3.4 microns plateau emission is shown to be a general spectral feature of vibrationally excited PAHs containing aliphatic hydrogens, especially those containing methyl groups; and finally, hot-band and overtone emissions arising from aromatic C-H vibrations are not observed in laboratory emission spectra, and therefore, in contrast to current assignments, are not expected to be observed in the UIRs. PMID- 11878351 TI - Medium modified two-body scattering amplitude from proton-nucleus total cross sections. AB - Recently (R.K. Tripathi, J.W. Wilson, F.A. Cucinotta, Nucl. Instr. and Meth. B 145 (1998) 277; R.K. Tripathi, F.A. Cucinotta, J.W. Wilson, NASA-TP-1998-208438), we have extracted nucleon-nucleon (N-N) cross-sections in the medium directly from experiment. The in-medium N-N cross-sections form the basic ingredients of several heavy-ion scattering approaches including the coupled-channel approach developed at the NASA Langley Research Center. Here, we investigate the ratio of real to imaginary part of the two-body scattering amplitude in the medium. These ratios are used in combination with the in-medium N-N cross-sections to calculate total proton-nucleus cross-sections. The agreement is excellent with the available experimental data. These cross-sections are needed for the radiation risk assessment of space missions. PMID- 11878350 TI - Chemistry of the organic-rich hot core G327.3-0.6. AB - We present gas-phase abundances of species found in the organic-rich hot core G327.3-0.6. The data were taken with the Swedish-ESO Submillimetre Telescope (SEST). The 1-3 mm spectrum of this source is dominated by emission features of nitrile species and saturated organics, with abundances greater than those found in many other hot cores, including Sgr B2 and OMC-1. Population diagram analysis indicates that many species (CH3CN, C2H3CN, C2H5CN, CH3OH, etc.) have hot components that originate in a compact (~2") region. Gas-phase chemical models cannot reproduce the high abundances of these molecules found in hot cores, and we suggest that they originate from processing and evaporation of icy grain mantle material. In addition, we report the first detection of vibrationally excited ethyl cyanide and the first detection of methyl mercaptan (CH3SH) outside the Galactic center. PMID- 11878352 TI - Medicare and TRICARE. AB - In 2001, two new health insurance programs called TRICARE for Life and the TRICARE Senior Pharmacy Program were established for armed services retirees and their dependents. In this brief, we discuss how these new programs work, who is eligible and how much they cost. We will also explore how the programs interact with your clients' Medicare benefits. PMID- 11878353 TI - Cratering rates on the Galilean satellites. AB - We exploit recent theoretical advances toward the origin and orbital evolution of comets and asteroids to obtain revised estimates for cratering rates in the jovian system. We find that most, probably more than 90%, of the craters on the Galilean satellites are caused by the impact of Jupiter-family comets (JFCs). These are comets with short periods, in generally low-inclination orbits, whose dynamics are dominated by Jupiter. Nearly isotropic comets (long period and Halley-type) contribute at the 1-10% level. Trojan asteroids might also be important at the 1-10% level; if they are important, they would be especially important for smaller craters. Main belt asteroids are currently unimportant, as each 20-km crater made on Ganymede implies the disruption of a 200-km diameter parental asteroid, a destruction rate far beyond the resources of today's asteroid belt. Twenty-kilometer diameter craters are made by kilometer-size impactors; such events occur on a Galilean satellite about once in a million years. The paucity of 20-km craters on Europa indicates that its surface is of order 10 Ma. Lightly cratered surfaces on Ganymede are nominally of order 0.5-1.0 Ga. The uncertainty in these estimates is about a factor of five. Callisto is old, probably more than 4 Ga. It is too heavily cratered to be accounted for by the current flux of JFCs. The lack of pronounced apex-antapex asymmetries on Ganymede may be compatible with crater equilibrium, but it is more easily understood as evidence for nonsynchronous rotation of an icy carapace. PMID- 11878354 TI - Observations of CH4, C2H6, and C2H2 in the stratosphere of Jupiter. AB - We have performed high-resolution spectral observations at mid-infrared wavelengths of CH4 (8.14 micrometers), C2H6 (12.16 micrometers), and C2H2 (13.45 micrometers) on Jupiter. These emission features probe the stratosphere of the planet and provide information on the carbon-based photochemical processes taking place in that region of the atmosphere. The observations were performed using our cryogenic echelle spectrometer CELESTE, in conjunction with the McMath-Pierce 1.5 m solar telescope between November 1994 and February 1995. We used the methane observations to derive the temperature profile of the jovian atmosphere in the 1 10 mbar region of the stratosphere. This profile was then used in conjunction with height-dependent mixing ratios of each hydrocarbon to determine global abundances for ethane and acetylene. The resulting mixing ratios are 3.9(+1.9)( 1.3) x 10(-6) for C2H6 (5 mbar pressure level), and 2.3 +/- 0.5 x 10(-8) for C2H2 (8 mbar pressure level), where the quoted uncertainties are derived from model variations in the temperature profile which match the methane observation uncertainties. PMID- 11878355 TI - The German Research Society (DFG) research project: new perspectives in transplantation medicine: the biological, medical, and ethical aspects of xenotransplantation and the derivation of organs from embryonic stem cells. PMID- 11878356 TI - Promoting marriage as welfare policy: looking at a public role in private lives. AB - This paper discusses the interest in and politics surrounding government's using welfare reform legislation to carve out a stronger role in promoting marriage. It examines trends in family formation, their impact on society, and the effect of single parenting, divorce, and step-parenting on child well-being. The paper also looks at the treatment of marriage in current government programs, new state activities to promote marriage, proposals for a stronger government role, and marriage experts' expectations for the success of marriage education programs. PMID- 11878357 TI - Red blood cell transfusion in critically ill patients. Emerging risks and alternatives. PMID- 11878360 TI - Australian triage nurses' decision-making and scope of practice. AB - A survey of 172 Australian triage nurses was undertaken to describe their scope of practice, educational background and to explore the self-reported influences perceived to impact on their decision-making. The survey results reveal variability in the educational requirements for nurses to triage. Indeed, over half of the nurses who participated in the study worked in emergency departments that provided no specified unit-based triage education. Additionally, substantial inter-respondent variations in nurses' self-reported participation in a range of decisions to expedite emergency care were identified. Analysis revealed significant associations between demographic characteristics of the triage service, levels of nurse' autonomy and the nurses' self-reported participation in a number of triage decisions. The findings of this study have implications for emergency nurse education and the development and evaluation of triage practice guidelines. PMID- 11878358 TI - A description of the adoption of the 'Fresh start' smoking cessation program by antenatal clinic managers. AB - This paper reports the dissemination by the Cancer Education Research Program (CERP) of a previously tested smoking cessation program called 'Fresh start' to 23 antenatal clinics. The program was specifically designed for use by staff in antenatal clinics. The aim of the study was to investigate the factors that influenced midwifery managers' adoption of the program. Clinics were randomly assigned to groups that received the program by simple dissemination (mail-out), or intensive dissemination (a mail-out, plus personal contact with midwifery facilitators). A case history approach was used to investigate the variables which influenced a midwifery manager's decision to adopt the program. The results indicated that intensive dissemination improved program adoption and that program components were selected to fit the physical and social context within antenatal clinics. Managers believed the main barriers to the implementation of the program were: the negative reactions of clients; insufficient time available for smoking cessation interventions; lack of support from professional colleagues; inability to provide follow-up to clients; staff turnover; and poor access and storage of materials. PMID- 11878359 TI - On-line alchemy--preventing gold being turned into lead! PMID- 11878362 TI - Relaxation: a nursing therapy to help relieve cardiac chest pain. AB - The experience of cardiac chest pain is always traumatic and stressful for patients. The available literature suggests that although nurses place a lot of importance on cardiac patients being pain free, it is apparent this is often not achieved. Research and documented case studies suggest that relaxation can play an important role in the treatment and prevention of this distressing symptom (Tiernan 1994; Altice and Jamison 1989; Guzzetta 1989). Nurses caring for patients with chest pain need to look beyond medical management and begin to challenge nursing practice to help patients deal effectively with chest pain in a way that meets each individual's needs. This article discusses ways in which relaxation, when used as an adjunct to medical therapies, can be a useful nursing management tool for effectively relieving cardiac pain. PMID- 11878361 TI - The role of the psychiatric consultation-liaison nurse in the general hospital. AB - The mainstreaming of psychiatric services has increased the amount of contact nurses have with clients experiencing mental health problems within the general hospital environment. A review of the literature suggests that general nurses find themselves lacking in the skills, confidence and knowledge to care adequately for these patients. The aim of this paper is to discuss the potential contribution of the psychiatric consultation-liaison nurse in addressing such problems in order to improve health outcomes for patients experiencing mental health problems. While a number of positions for Psychiatric Consultation-Liaison Nurses are being created throughout Australia, there is a paucity of literature relating to the development of this important role. This paper is intended to contribute to the advancement of a body of knowledge in this area. PMID- 11878363 TI - 'They' say it's the best health system in the world..... PMID- 11878364 TI - Birth centre or labour ward? A comparison of the clinical outcomes of low-risk women in a NSW hospital. AB - A number of birth centres were established in New South Wales as a result of the Shearman Report (NSW Health Department 1989). The objective of this study was to compare the obstetric outcomes, primarily caesarean section rates, of low-risk women presenting in spontaneous labour to the birth centre with those attending the hospital's conventional labour ward. The study showed that there was no significant difference in the caesarean section rate between the groups (3.5% in the birth centre and 4.3% in the labour ward). We suggest that the site of birthing does not affect clinical outcomes for low-risk women at this hospital. These results are relevant to contemporary clinical practice as they question the basis upon which birth centres have been popularised, that is, the medicalisation of birth in conventional labour wards increases intervention rates. PMID- 11878365 TI - Getting serious about sustainability. AB - Sustainability and sustainable development are catchwords that dominate today's environmental science and policy discourse. It is easy to demonstrate that most of the activities of today's industrial society are unsustainable. Unfortunately, much of the talk about sustainability lacks a basic understanding of what truly sustainable activity would be. To set sustainability as a target or goal for our industrial society, we must be able to quantify that target or goal. We propose four basic steps to begin this process for one aspect of sustainability, the rate of use of resources: (i) establish the available supply of the chosen resource; (ii) allocate the annual permissible supply according to a reasonable formula or market process; (iii) establish the "recaptureable" resource base; and (iv) derive the sustainable limiting rate of use and compare to the current rate of use. We apply these sustainability measurement methods to three common materials in industrial society: zinc, germanium, and greenhouse gases. These examples demonstrate that with some basic (although potentially controversial) assumptions, quantitative sustainable use goals can be set and current performance relative to those goals can be evaluated. The assumptions and approximates we have used are meant to stimulate thought and debate, beginning a long conversation on the measurement of sustainability. PMID- 11878366 TI - Current directions in the practice of environmental risk assessment in the United Kingdom. AB - The manner in which regulators apply environmental risk assessment to their decisions on managing risk is changing. Expectations of risk assessment work are becoming clearer, the social issues agenda is having an impact on risk assessment practice, and there is a trend toward harmonizing approaches to the treatment of environmental risk. For risk analysts, the multiplicity of environmental problems is providing opportunities for the transfer of expertise between the different contexts of applying environmental risk assessment. With the latter as a focus, we summarize recent policy developments in the United Kingdom and illustrate how Government guidance on environmental risk assessment and management is being implemented. We emphasize the need for proportionality in risk analysis, the targeting of regulatory effort to risk, and the explicit treatment of uncertainty. These developments are contributing toward better "risk-informed" environmental decisions in which risk analysis plays an important part alongside other considerations. The forward agenda is likely to see further practical integration between technical risk issues and economic and social concerns, and the positioning of environmental risk assessment within a broader landscape of decision-making tools. PMID- 11878367 TI - Emission inventories of NOx from commercial energy consumption in China, 1995 1998. AB - Emissions of nitrogen oxides (NOx) in China are of great concern because of their impact on local air pollution as well as on regional environmental risks such as acid rain, eutrophication, tropospheric ozone, fine particulate matter, and loss of biodiversity. In this paper, total anthropogenic emissions of NOx in China during the past two decades are estimated on the basis of commercial energy consumption and NOx emission factors of different sectors and fuel types. Specifically, emission inventories of NOx from 31 provinces, 7 economic sectors, and 11 fuel types from 1995 to 1998 are evaluated and analyzed in detail, and the distribution of provincial-based average NOx emission intensity in 1998 is presented. It can be seen that the calculated national total emissions of NOx have rapidly increased from 4.76 Mt in 1980 to a peak value of about 12.03 Mt in 1996, with an average annual growth rate of 6%. However, instead of increasing, NOx emissions in 1997 and 1998 decreased by 0.37 Mt and 0.85 Mt, respectively, as compared to the peak in 1996. In short, NOx emissions caused by energy use are mainly concentrated in the more populated and industrialized areas of China, i.e., the Eastern Central and Southeastern areas. These results imply that China's NOx emissions in the future may not become as high as previously expected which will be helpful for policy making to control local NOx pollution as well as to reduce acid rain and tropospheric ozone in East Asia. PMID- 11878368 TI - R-134a emissions from vehicles. AB - We report the first study of R-134a (also known as HFC-134a and CF3CFH2) refrigerant leakage from air conditioning (AC) systems of modern vehicles. Twenty eight light duty vehicles from five manufacturers (Ford, Toyota, Daimler Chrysler, General Motors, and Honda) were tested according to the USEPA (Federal) extended diurnal test procedure using the Sealed Housing for Evaporative Determination (SHED) apparatus. All tests were conducted using stationary vehicles with the motor and air conditioning system turned off. R-134a was measured using gas chromatography (GC) with a flame ionization detector (FID). All vehicles exhibited measurable R-134a leakage over the 2-day diurnal test. Leak rates of R-134a ranged from 0.01 to 0.36 g/day with an average of 0.07+/ 0.07 g/day. When combined with leakage associated with vehicle operation, servicing, and disposal we estimate that the lifetime average R-134a emission rate from an AC equipped vehicle is 0.41+/-0.27 g/day (the majority of emissions are associated with vehicle servicing and disposal). Assuming that the average vehicle travels 10 000 miles per year we estimate that the global warming impact of R-134a leakage from an AC equipped vehicle is approximately 4-5% of that of the CO2 emitted by the vehicle. The results are discussed with respect to the contribution of vehicle emissions to global climate change. PMID- 11878369 TI - Salt marsh recovery and oil spill remediation after in-situ burning: effects of water depth and burn duration. AB - Effects of water depth, burn duration, and diesel fuel concentration on the relationship between recovery of marsh vegetation, soil temperature, and oil remediation during in-situ burning of oiled mesocosms were investigated. The water depth over the soil surface during in-situ burning was a major factor controlling recovery of the salt marsh grass, Spartina alterniflora. Ten centimeters of water overlying the soil surface was sufficient to protect the marsh soil from burn impacts with soil temperatures <37 degrees C and high plant survival rate. In contrast, a water table 10 cm below the soil surface resulted in mean soil temperatures > 100 degrees C at the 2-cm soil depth, which completely inhibited the post-burn recovery of S. alterniflora. Although poor plant recovery was also apparent in the treatments with 0 and 2 cm of water over the soil surface, this result was likely due to the chemical stress of the diesel fuel used to create the fire rather than the heat, per se, which never reached the estimated lethal temperature of 60 degrees C. In-situ burning effectively removed more than 95% of floating oil from the water surface. Thus, in-situ burning prevented the oil from potentially contaminating adjacent habitats. However, in-situ burning did not effectively remediate the oil that had penetrated the soil. PMID- 11878370 TI - Modeling silica sorption to iron hydroxide. AB - Experiments were conducted to investigate the fundamentals of silica sorption onto preformed ferric hydroxide at pH 5.0-9.5 and silica concentrations of 0-200 mg/L as SiO2. At all pHs studied, sorption densities exceeding monolayer sorption were observed at silica levels typical of natural waters. Under some circumstances, sorption equaled or exceeded a monolayer while the particle zeta potential remained positive, a phenomenon that is completely inconsistent with available surface complexation models. To address this deficiency, an extended surface complexation model was formulated in which soluble dimeric silica (i.e., Si2O2(OH)5-) sorbs directly to iron surface sites. This new model fit sorption density data up to 0.40 mol SiO2/mol Fe and accurately predicted trends in zeta potential and observed H+ release during silica sorption to ferric hydroxide. PMID- 11878371 TI - Formation of N-nitrosodimethylamine (NDMA) from dimethylamine during chlorination. AB - Chlorine disinfection of secondary wastewater effluent and drinking water can result in the production of the potent carcinogen N-nitrosodimethylamine (NDMA) at concentrations of approximately 100 and 10 parts per trillion (ng/L), respectively. Laboratory experiments with potential NDMA precursors indicate that NDMA formation can form during the chlorination of dimethylamine and other secondary amines. The formation of NDMA during chlorination may involve the slow formation of 1,1-dimethylhydrazine by the reaction of monochloramine and dimethylamine followed by its rapid oxidation to NDMA and other products including dimethylcyanamide and dimethylformamide. Other pathways also lead to NDMA formation during chlorination such as the reaction of sodium hypochlorite with dimethylamine. However, the rate of NDMA formation is approximately an order of magnitude slower than that observed when monochloramine reacts with dimethylamine. The reaction exhibits a strong pH dependence due to competing reactions. It may be possible to reduce NDMA formation during chlorination by removing ammonia prior to chlorination, by breakpoint chlorination, or by avoidance of the use of monochloramine for drinking water disinfection. PMID- 11878372 TI - Peroxidase-catalyzed oxidative coupling of phenols in the presence of geosorbents: rates of non-extractable product formation. AB - Oxidative coupling processes in subsurface systems comprise a form of natural contaminant attenuation in which hydroxylated aromatic compounds (HACs) are incorporated into soil/sediment organic matter matrices. Here we describe the oxidative coupling of phenol catalyzed by horseradish peroxidase (HRP) in systems containing two geosorbents having organic matter of different composition; specifically Chelsea soil, a near-surface geologically young soil having a predominantly humic-type soil/sediment organic matter (SOM) matrix, and Lachine shale, a diagenetically older natural material having a predominantly kerogen type SOM matrix. It was found that each of these two different types of natural geosorbents increased the formation of non-extractable coupling products (NEPs) over that which occurred in solids-free systems. The extent of coupling was higher in the systems containing humic-type Chelsea SOM than in those containing kerogen-type Lachine SOM. It was observed that HRP inactivation by free radical attack was significantly reduced in the presence of each geosorbent. A rate model was developed to facilitate quantitative evaluation and mechanistic interpretation of such coupling processes. Experimental rate measurements revealed thatthe greater extent of reaction observed in the presence of Chelsea soil than in the presence of Lachine shale can be attributed to two factors: (i) more effective protection of HRP from inactivation by the Chelsea SOM and (ii) the greater reactivity of Chelsea SOM with respect to cross-coupling. Interrelationships among enzyme protection, cross-coupling reactivity, and SOM chemistry are discussed. PMID- 11878373 TI - Interaction of methyl bromide with soil. AB - Because methyl bromide (CH3Br) is a widely used agricultural fumigant for soil disinfection, it is important to know the chemical behavior and fate of CH3Br as a result of its use for soil treatment. A solid-state 13C NMR study of 13CH3Br treated soil and soil-component samples shows that methylation of soil organic matter may be the major pathway for degradation of CH3Br in soils. Adsorption of CH3Br on a dried clay like Ca-montmorillonite or kaolinite does not contribute directly to the degradation of CH3Br. The results are interpreted in terms of the chemical structures of separated soil fractions and the nature of the separation procedure. PMID- 11878374 TI - Specific UV absorbance of Aldrich humic acid: changes during transport in aquifer sediment. AB - This study examined the transport behavior of Aldrich humic acid (AHA) in low natural organic carbon content sediment contaminated with tetrachloroethene (PCE), for comparison to a nonionic surfactant mixture previously examined in the same system. Tracking of individual molecular weight (MW) fractions of AHA was attempted by UV absorbance, followed by conversion to mass of carbon using specific ultraviolet absorbance (SUVA) (UV absorbance per mass of carbon) measurements. The analysis required determination of variations of SUVA with MW, which showed a maximum at 10 000 Daltons. Furthermore, SUVAs of AHA MW fractions greater than about 10 000 Daltons increased following AHA interaction with sediment in batch experiments, and this was associated with AHA-driven leaching of cations from the sediment. AHA transport was examined in a series of three columns representing the up-gradient, residual-zone, and down-gradient portions of a DNAPL contaminated site. SUVAs of larger MW AHA fractions cycled through decreased, increased, and eventual return to influent values during the early, intermediate, and final stages of breakthrough, respectively. These variations were attributable to a combination of preferential adsorption of low MW fractions of the AHA during early breakthrough and AHA-driven leaching of sediment cations during intermediate breakthrough, with eventual exhaustion of sediment cation complexation during the final stage of breakthrough. The complex variations in SUVA precluded accurate conversion of measured UV absorbance to carbon mass. However, the effect of AHA loss to sediment on the solubilizing capacity of the AHA solution was indicated by the breakthrough behavior of AHA-solubilized PCE, which showed that AHA loss from the aqueous phase during transport in this system did not decrease the solubilizing capacity of the AHA mixture. PMID- 11878375 TI - Electrochemical properties of natural organic matter (NOM), fractions of NOM, and model biogeochemical electron shuttles. AB - In this study, cyclic voltammetry was used to characterize the redox properties of natural organic matter (NOM). Using a stationary platinum working electrode, minimal concentrations of electrolyte, and dimethyl sulfoxide (DMSO) as the solvent, we were able to resolve two pairs of oxidation and reduction peaks for a fraction of Georgetown NOM that is enriched in polyphenolic moieties (NOM-PP). Applying our method to other fractions of Georgetown NOM, and to samples of NOM from a wide range of other sources, gave cyclic voltammograms (CVs) that generally contained fewer distinguishing features than those obtained with NOM PP. For comparison, CVs were also obtained using our method on six quinone model compounds: anthraquinone-2,6-disulfonate (AQDS), lawsone, juglone, menadione, menaquinone-4, and ubiquinone-5. The CVs of these quinones were similar in shape to the CV of NOM-PP, consistent with the notion that quinones are the dominant redox-active moieties associated with NOM. Quantitative analysis of the peaks in these CVs showed that the peak potentials (Ep) were separated by more than 0.059 V and that the peak currents (i(p)) were linearly related to the square root of the scan rate (v0.5) and concentration (C) for both NOM-PP and the model quinones. Equivalent results were obtained with a rotating Pt disk electrode. From this we conclude that NOM-PP and the model quinones undergo similar sequences of two one-electron, quasi-reversible, diffusion controlled, electron transfers at the Pt electrode surface in DMSO. Although it is difficult to relate these results to Nernstian standard potentials vs the standard hydrogen electrode (SHE) under aqueous conditions, it is clear that the apparent formal potential for NOM-PP lies between the corresponding potentials for menadione and juglone and well above that of AQDS. Attempts to derive correlations between Ep and i(p) for the NOMs with quantifiable electrode response and other measurable properties of NOM (including trace metal content and UV-vis absorbance) did not yield any strong relationships. PMID- 11878376 TI - Products and mechanism of the reaction of OH radicals with 2,2,4-trimethylpentane in the presence of NO. AB - Alkanes are important constituents of gasoline fuel and vehicle exhaust, with branched alkanes comprising a significant fraction of the total alkanes observed in urban areas. Products of the gas-phase reactions of OH radicals with 2,2,4 trimethylpentane and 2,2,4-trimethylpentane-d18 in the presence of NO at 298+/-2 K and atmospheric pressure of air have been investigated using gas chromatography with flame ionization detection (GC-FID), combined gas chromatography-mass spectrometry (GC-MS), and in situ atmospheric pressure ionization tandem mass spectrometry (API-MS). Acetone, 2-methylpropanal, and 4-hydroxy-4-methyl-2 pentanone were identified and quantified by GC-FID from 2,2,4-trimethylpentane with molar formation yields of 54+/-7%, 26+/-3%, and 5.1+/-0.6%, respectively; upper limits to the formation yields of acetaldehyde, 2,2-dimethylpropanal, and 4,4-dimethyl-2-pentanone were obtained. Additional products observed from 2,2,4 trimethylpentane by API-MS and API-MS/MS analyses using positive and negative ion modes were hydroxy products of molecular weight 130 and 144, a product of molecular weight 128 (attributed to a C8-carbonyl), and hydroxynitrates of molecular weight 135, 177, and 191 (attributed to HOC4H8ONO2, HOC7H14ONO2, and HOC8H16-ONO2, respectively). Formation of HOC8H16ONO2 and HOC7H14-ONO2 is consistent with the observation of products of molecular weight 207 (HOC8D16ONO2) and 191 (HOC7D14-ONO2), respectively, in the API-MS analyses of the 2,2,4 trimethylpentane-d18 reaction (-OD groups rapidly exchange to -OH groups under our experimental conditions). These product data allow the reaction pathways to be delineated to a reasonable extent, and the reaction mechanism is discussed. PMID- 11878377 TI - Insights into the formation and degradation mechanisms of methylenedinitramine during the incubation of RDX with anaerobic sludge. AB - In an earlier study, we reported that hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) biodegraded with domestic anaerobic sludge to produce a key RDX ring cleavage intermediate that was tentatively identified as methylenedinitramine (O2NNHCH2NHNO2) using LC/MS with negative electrospray ionization (ES-). Recently, we obtained a standard material of methylenedinitramine and thus were able to confirm its formation as the key initial RDX intermediate. In water alone or in the presence of sludge, methylenedinitramine decomposed to N20 and HCHO. Only in the presence of sludge HCHO converted further to carbon dioxide. To test our hypothesis that water was involved in the formation of methylenedinitramine during incubation of RDX with sludge, we allowed the energetic compound to biodegrade in several D2O/H2O solutions (90, 50, and 0% v/v). We observed three distinctive deprotonated or dedeuterated mass ions at 135, 136, and 137 Da that were attributed to the formation of nondeuterated (H-methylenedinitramine), monodeuterated (D1-methylenedinitramine), and dideuterated methylenedinitramine (D2-methylenedinitramine), respectively. Two controls were prepared in D2O both in the absence of sludge; the first contained methylenedinitramine, and the second contained RDX. Neither control produced any deuterated methylenedinitramine, thus excluding the occurrence of any abiotic D/H exchange between D2O and either methylenedinitramine or RDX. The results supported the occurrence of an initial enzymatic reaction on RDX, yet they did not provide compelling evidence on whether methylenedinitramine was an initial RDX enzymatic hydrolysis product or simply formed via the spontaneous hydrolysis of an anonymous initial RDX enzymatic product. PMID- 11878378 TI - Mechanisms of hydrogen peroxide decomposition in soils. AB - The rates and mechanisms of hydrogen peroxide (H2O2) decomposition were examined in a series of soil suspensions at H2O2 concentrations comparable to those found in rainwaters. The formation of hydroxyl radical (OH) as a possible decomposition intermediate was investigated using a new, highly sensitive method. In surface soils with higher organic matter or manganese content, H2O2 usually decayed rapidly, with disproportionation to water and dioxygen dominating the decomposition, whereas the formation of the hydroxyl radical (OH) represented <10% of the total H2O2 decomposed. In contrast, for soils with lower organic matter content, H2O2 usually decayed much more slowly, but OH was a major product of the H2O2 decomposed. The decomposition was principally associated with soil particles, not the soil supernatant. Different sterilization techniques indicated that decomposition of H2O2 was at least partly due to biological activity. Because the loss of H2O2 can largely be accommodated by the production of O2 and OH within these soils, our results suggest that disproportionation through a catalase-type mechanism and the production of OH through a Haber-Weiss mechanism represent the principal routes through which H2O2 is lost. PMID- 11878379 TI - Reduction of the pesticides oxamyl and methomyl by FeII: effect of pH and inorganic ligands. AB - This work examines the effect that pH and selected inorganic ligands have on the kinetics of reactions between FeII and two structurally related oxime carbamate pesticides, oxamyl and methomyl. In anoxic solutions containing FeII, these compounds degrade by parallel elimination and reduction pathways. Rates of FeII independent carbamate elimination (EIcb mechanism) are proportional to [OH-], increasing 10-fold for each unit increase in pH. In homogeneous solution, rates of carbamate reduction by 0.5 mM FeII are relatively constant at pH <7, but increase dramatically between pH 7 and pH 8.3. At pH >8.3, Fe(OH)2(s) precipitation occurs, and carbamates react with both solution-phase and solid phase FeII. Carbamate reduction by FeII is not significantly affected by the presence of chloride, bromide, nitrate, perchlorate, and sulfate. In contrast, increased rates of carbamate reduction are observed in solutions containing fluoride, carbonate, and phosphate. Kinetic measurements are interpreted in terms of changing FeII speciation according to the expression kred = [FeII]sigmaikialphai, where k(red) is the pseudo-first-order rate constant for carbamate reduction, [FeII] is the total FeII concentration, and ki and alphai are the second-order rate constant and fractional concentration of each FeII species, respectively. It follows that the overall kinetics of carbamate reduction is a function of the identity and concentration of individual FeII species present in solution as well as the inherent reactivity of each species with carbamates. The magnitude of ki is related to the standard one-electron reduction potential (E(H) degrees) of the corresponding FeIII/FeII redox couple. PMID- 11878380 TI - Modeling the kinetics of ferrous iron oxidation by monochloramine. AB - The maintenance of disinfectants in distribution systems is necessary to ensure drinking water safety. Reactions with oxidizable species can however lead to undesirable disinfectant losses. Previous work has shown that the presence of Fe(II) can cause monochloramine loss in distribution system waters. This paper further examines these reactions and presents a reaction mechanism and kinetic model. The mechanism includes both aqueous-phase reactions and surface-catalyzed reactions involving the iron oxide product. In addition, it considers competitive reactions involving the amidogen radical that lead to a nonelementary stoichiometry. Using the method of initial rates, the aqueous-phase reactions were found to have first-order dependencies on Fe(II), NH2Cl, and OH- and a rate coefficient (kNH2Cl,soln) of 3.10 (+/-0.560) x 10(9) M(-2) min(-1). The surface mediated reactions were modeled by assuming the formation of two surface species: >FeOFe+ and >FeOFeOH. Using numerical techniques, combined rate coefficients for the surface-mediated processes were determined to be 0.56 M(-3) min(-1) and 3.5 x 10(-18) M(-4) min(-1), respectively. The model was then used to examine monochloramine and Fe(II) stability under conditions similar to those observed in distribution systems. Our findings suggest the potential utility of monochloramine as an oxidant for Fe(III) removal in drinking water treatment. PMID- 11878381 TI - New processes in the environmental chemistry of nitrite: nitration of phenol upon nitrite photoinduced oxidation. AB - The role of nitrite as an environmental factor has been widely recognized. Nitrite is a relevant source of *OH in the atmosphere, both in the gas phase via photolysis of gaseous HNO2 and in atmospheric hydrometeors by photolysis of NO2-. In aqueous systems, *OH production through nitrite photolysis can be negligible due to the competition for light absorption by dissolved Fe(III), colloidal iron oxides, and nitrate. These photoexcited oxidants interact with NO2- and HNO2 to form *NO2, either directly or via formation of *OH. As a consequence, nitrite and nitrous acid may act as *NO2 rather than *OH sources. The radical *NO2 is involved in the nitration of many aromatic compounds, of which phenol is a model in this work. Kinetic measurements using 2-propanol as *OH scavenger show that the direct production of *OH by aqueous Fe(III) species decreases as pH increases. At slightly acidic and neutral pH values, oxidation of nitrite occurs by direct electron transfer to photoexcited Fe(III)aq species or colloidal iron oxides, in addition to the *OH-mediated oxidation of NO2-. The reported findings suggest a completely new role of nitrite in aquatic environments. PMID- 11878382 TI - Significance of semivolatile diesel exhaust organics for secondary HONO formation. AB - The atmospheric origin of nitrous acid (HONO) is largely unknown despite its estimated importance as an OH source during daytime due to its rapid photolysis. Recently, primary HONO contained in automobile exhaust as well as secondary HONO formation on soot particles have been invoked as possible HONO sources, but none of them is able to account for the observed HONO to NOx ratios of up to 0.04 in the atmosphere. In this paper, we show that semivolatile and/or water-soluble species contained in diesel exhaust are significantly involved in secondary HONO formation. These species are not associated with soot when the exhaust exits the tailpipe. To quantify these species and to assess the reaction kinetics leading to HONO, experiments were performed in which filtered but hot diesel exhaust gas interacted with a glass surface as well as a water film mimicking dry and wet surfaces to which exhaust might be exposed. A fraction of 0.023 of the NOx emitted was heterogeneously converted to HONO, which is at least three times more than the primary HONO emissions by diesel engines and a fraction of 50 larger than HONO formed on diesel soot particles that do not contain the semivolatile organics. PMID- 11878383 TI - Characterization of the atrazine's bound (nonextractable) residues using fractionation techniques for soil organic matter. AB - Atrazine's bound residues (BR), in the range of 10-40% of the applied atrazine,were obtained by laboratory incubation (56 d) of four soils having different capacities to degrade atrazine in relation to the presence or absence of a microflora able to mineralize the triazinic ring. Soil size fractionation followed by alkaline extraction, before and after HF treatment, and then acid hydrolysis with 2 M HCl in reflux conditions was applied to the soils containing BR. Most of the BR were in the finest fraction (<20 microm) that contained the humified organic matter (from 61 to 77% of the total BR), and between 78 and 89% was made soluble during the different steps of the chemical fractionation procedure. From 20 to 50% of the BR of the fraction <20 microm was identified as the intact atrazine and its main derivatives, indicating that this proportion of the BR was probably formed by entrapment in voids of the soil organic matter. Between 13 and 30% of the BR was associated to humic acids (HA); they were not dialyzable and were released by acid hydrolysis with HCl, indicating that these BR were chemically bound to HA by an heteroatomic bond after the substitution of the chlorine atom of atrazine. Comparison of the results obtained for the four soils indicated that (i) an important activity of microorganisms able to mineralize the triazinic ring favors the formation of highly degraded products that can form BR; (ii) a soil pH <6 favors the formation and stabilization of hydroxylated derivatives of atrazine, and (iii) a high content of humic acids favors the formation of chemically bound residues. PMID- 11878384 TI - Association of methylmercury with dissolved humic acids. AB - Sorption of methylmercury (MeHg) to three different humic acids was investigated as a function of pH and humic concentration. The extent of sorption did not show a strong pH dependence within the pH range of 5-9. Below pH 5, a decrease in adsorption for all humic samples was observed. The experimental data for equilibrium sorption of MeHg were modeled using a discrete log K spectrum approach with three weakly acidic functional groups. The modeling parameters, which were the equilibrium binding constants and the total binding capacities, represented the data well at all MeHg and humic concentrations and pH values for a given humic sample. The estimated binding constants for complexes of MeHg with humic acids were similar in magnitude to those of MeHg with thiol-containing compounds, suggesting that binding of MeHg involves the thiol groups of humic acids. The results show that only a small fraction of the reduced sulfur species in humic substances may take part in binding MeHg, but in most natural systems, this subfraction is considerably higher in concentration than ambient MeHg. The model developed here can be incorporated into speciation models to assess the bioavailability of MeHg in the presence of dissolved organic matter and competing ligands such as chloride and sulfide. PMID- 11878385 TI - Anaerobic biotransformation of tetrabromobisphenol A, tetrachlorobisphenol A, and bisphenol A in estuarine sediments. AB - Biotransformation of the flame retardants tetrabromobisphenol A and tetrachlorobisphenol A, and their ultimate biodehalogenation product, bisphenol A, was examined in anoxic estuarine sediments. Dehalogenation of tetrabromobisphenol A and tetrachlorobisphenol A was examined under conditions promoting either methanogenesis or sulfate reduction as the primary terminal electron-accepting process. Complete dehalogenation of tetrabromobisphenol A to bisphenol A with no further degradation of bisphenol A, was observed under both methanogenic and sulfate-reducing conditions. Dehalogenation of tetrachlorobisphenol A under both methanogenic and sulfate-reducing conditions resulted in the accumulation of a persistent dichlorinated bisphenol A isomer, while no bisphenol A was formed. Co-amendment of sediment enrichments with either 2,6-dibromo- or 2,6-dichlorophenol did not affect the extent of dehalogenation as compared to sediments that were amended only with the flame retardants. Sediment cultures pre-acclimated on 2-bromophenol dehalogenated the flame retardants in a manner similar to that of fresh sediments. No loss of bisphenol A was observed in separate incubations within 162 days under conditions promoting either methanogenesis, sulfate-reduction, iron(III)-reduction, or nitrate-reduction. Furthermore, identical enrichments that readily degraded 4-hydroxybenzoate, a structural analogue of bisphenol A, did not exhibit bisphenol A degradation. The dehalogenation of tetrabromo- and tetrachlorobisphenol A and the potential for accumulation of bisphenol A in anoxic sediments is significant given the widespread use of these chemicals. PMID- 11878387 TI - Predicting the emission rate of volatile organic compounds from vinyl flooring. AB - A model for predicting the rate at which a volatile organic compound (VOC) is emitted from a diffusion-controlled material is validated for three contaminants (n-pentadecane, n-tetradecane, and phenol) found in vinyl flooring (VF). Model parameters are the initial VOC concentration in the material phase (C0), the material/air partition coefficient (K), and the material-phase diffusion coefficient (D). The model was verified by comparing predicted gas-phase concentrations to data obtained during small-scale chamber tests and by comparing predicted material-phase concentrations to those measured at the conclusion of the chamber tests. Chamber tests were conducted with the VF placed top-side-up and bottom-side-up. With the exception of phenol and within the limits of experimental precision, the mass of VOCs recovered in the gas-phase balances the mass emitted from the material phase. The model parameters (C0, K, and D) were measured using procedures completely independent of the chamber test. Gas- and material-phase predictions compare well to the bottom-side-up chamber data. The lower emission rates for the top-side-up orientation may be explained by the presence of a low-permeability surface layer. The sink effect of the stainless steel chamber surface was shown to be negligible. PMID- 11878386 TI - Benzothiazolamines as tire-derived molecular markers: sorptive behavior in street runoff and application to source apportioning. AB - Wash-off and sorptive behaviors of two benzothiazolamines (BTs) [i.e., 2-(4 morpholinyl)benzothiazole (24MoBT) and N-cyclohexyl-2-benzothiazolamine (NCBA)] have been investigated as possible molecular markersfortire debris and/or road dust transported in highway runoff water. Sum of dissolved and particulate 24MoBT and NCBA concentrations in runoff water ranged from 15 to 417ng/L and from 22to 508ng/L, respectively. Proportions of NCBA in particulate (>0.7microm) phase (<9 79%) were larger than that of 24MoBT (<1-14%), which was consistent with their experimentally determined octanol/water partition coefficients (Kow; 10(4.23+/ 0.14) for NCBA; 10(2.42+/-0.03) for 24MoBT). The organic carbon-normalized in situ partition coefficient (Koc') observed in runoff events (10(4.69+/-0.28) for NCBA; 10(3.42+/-0.23) for 24MoBT) were 1 order of magnitude higher than those expected from their Kow, indicating strong affinity of BTs to suspended particulate matter (SPM) in runoff water. Furthermore, in desorption experiments lasting 24 h, we observed almost the same levels of Koc' as those in runoff events, implying that significant fractions of BTs are strongly associated with runoff particles and not easily available to equilibrium partitioning. NCBA was ubiquitous in sediments from the Nogawa River receiving runoff from the Chuo Highway, whereas many of those samples had undetectable levels of 24MoBT. All of above results indicate that NCBA would be more suitable than 24MoBT as a molecular marker for runoff particles loading the aquatic environment. By using SPM-weighted mean concentration of particulate NCBA, at least 3.3+/-1.6% of the mass in the Nogawa sediments is estimated to be from runoff SPM. PMID- 11878388 TI - Automatic buffer capacity model building for advanced interpretation of titration curves. AB - An automatic buffer capacity model building algorithm was developed. The objective of this algorithm is to characterize multiple buffer systems from experimental titration curves. Starting from a simple mathematical model that incorporates the available prior knowledge on the buffer system, the model is sequentially extended by incorporating additional monoprotic buffer equations. To select the most appropriate model from the resulting set of automatically built models, a model structure selection technique was constructed from existing methods. PMID- 11878389 TI - Hydrogen isotopic composition of individual n-alkanes as an intrinsic tracer for bioremediation and source identification of petroleum contamination. AB - The isotopic signatures of crude oil hydrocarbons are potentially powerful intrinsic tracers to their origins and the processes by which the oils are modified in the environment. Stable carbon isotopic data are of limited use for studying petroleum contaminants because of the relatively small amount of isotopic fractionation that occurs during natural processes. Hydrogen isotopes, in contrast, are commonly fractionated to a much greater extent and as a result display larger variations in delta values. We studied the effect of in vitro aerobic biodegradation on the hydrogen isotopic composition of individual n alkanes from crude oil. The isotopic analysis was conducted using gas chromatography-thermal conversion-isotope ratio mass spectrometry. In general, biodegradation rates decreased with increasing hydrocarbon chain length, consistent with previous studies. More importantly the n-alkanes that were degraded at the fastest rates (n-C15 to n-C18) also showed the largest overall isotopic fractionation (approximately 12-25 per thousand deuterium enrichment), suggesting that the lower molecular weight n-alkanes can be used to monitor in situ bioremediation of crude oil contamination. The hydrogen isotopic compositions of the longer chain alkanes (n-C19 to n-C27) were relatively stable during biodegradation (<5%o overall deuterium enrichment), indicating that these compounds are effective tracers for oil-source identification studies. PMID- 11878390 TI - Corn leaf nitrate reductase--a nontoxic alternative to cadmium for photometric nitrate determinations in water samples by air-segmented continuous-flow analysis. AB - Development, characterization, and operational details of an enzymatic, air segmented continuous-flow analytical method for colorimetric determination of nitrate + nitrite in natural-water samples is described. This method is similar to U.S. Environmental Protection Agency method 353.2 and U.S. Geological Survey method 1-2545-90 except that nitrate is reduced to nitrite by soluble nitrate reductase (NaR, EC 1.6.6.1) purified from corn leaves rather than a packed-bed cadmium reactor. A three-channel, air-segmented continuous-flow analyzer configured for simultaneous determination of nitrite (0.020-1.000 mg-N/L) and nitrate + nitrite (0.05-5.00 mg-N/L) by the nitrate reductase and cadmium reduction methods-was used to characterize analytical performance of the enzymatic reduction method. At a sampling rate of 90 h(-1), sample interaction was less than 1% for all three methods. Method detection limits were 0.001 mg of NO2- -N/L for nitrite, 0.003 mg of NO3-+ NO2- -N/L for nitrate + nitrite by the cadmium-reduction method, and 0.006 mg of NO3- + NO2- -N/L for nitrate + nitrite bythe enzymatic-reduction method. Reduction of nitrate to nitrite by both methods was greater than 95% complete overthe entire calibration range. The difference between the means of nitrate + nitrite concentrations in 124 natural-water samples determined simultaneously bythe two methods was not significantly different from zero at the p = 0.05 level. PMID- 11878391 TI - Spatial applicability of emission factors for modeling mobile emissions. AB - Driving cycles are used to create mobile emission factors. Emission factors are, in turn, used to estimate mobile source inventories, provide standards for new vehicle emissions testing, and facilitate comparisons of laboratory experiments. This study examines the spatial representativeness of the driving cycles underlying California's CO, THC, and NOx emission rates that are applied when estimating regional mobile emissions inventories. Sixteen randomly selected vehicles were tested on a laboratory dynamometer using driving cycles representative of driving in different cities. A total of 214 tests, with repetitions, representing six driving cycles, were conducted on the 16 vehicles. We used a random effects analysis of variance to statistically examine the differences in the resulting emission rates. The study results suggest that California mobile source pollutant inventories prepared using emission rates based on the standard drive cycle may be off by as much as 30% for regions where traffic congestion and roadway networks differ significantly from those of Los Angeles. PMID- 11878392 TI - Fluorescence inner-filtering correction for determining the humification index of dissolved organic matter. AB - The use of fluorescence spectrometry has been suggested as a simple method to determine the extent of natural organic matter humification by quantifying the red-shifting of fluorescence emission that occurs with increasing humification. Humification indices are calculated by dividing fluorescence intensity at longer wavelengths by intensity at shorter wavelengths. These indices calculated without any specific efforts to standardize dissolved organic matter (DOM) concentration will result in index values thatvary with DOM concentration due to fluorescence innerfiltering effects. This study critically evaluated the effect of DOM concentration on humification index determination using organic matter isolated from field corn extract, soil: water extract, and soil fulvic acid. The results show that humification index values are sensitive to DOM concentration of the solution and are linear with respect to transmittance of the solution at the 254 nm used as the excitation wavelength. An approximate correction for DOM is to exploit the linear nature of the regression fit and to determine index values at the extrapolated 100% transmittance value. An exact correction using explicit correction factors for both primary and secondary innerfiltration effects was shown to give humification index values that are concentration invariant when absorbance of the solution at 254 nm was less than approximately 0.3 unit. Defining the humification index as the fluorescence intensity in the 300-345 nm region divided by the sum of intensity in the 300-345 nm and 435-480 nm regions was statistically advantageous. This study suggests that for quantitative results which can be used to compare humification of natural organic matter across different studies, correction of the fluorescence emission spectra for innerfiltration effects is needed. PMID- 11878393 TI - Improved method for quantifying levoglucosan and related monosaccharide anhydrides in atmospheric aerosols and application to samples from urban and tropical locations. AB - An improved analytical method was developed and validated for the determination of the monosaccharide anhydrides levoglucosan, mannosan, and galactosan in atmospheric aerosol samples. The method uses an external recovery standard, extraction in dichloromethane, trimethylsilylation, addition of an internal standard (1-phenyl dodecane), and analysis by gas chromatography with flame ionization detection (GC-FID) and gas chromatography/mass spectrometry (GC/MS). As external recovery standard, we selected 1,2,3-trihydroxyhexane, which has a similar polarity as the monosaccharide anhydrides; furthermore, it was ensured that the trimethylsilylation step leads to complete derivatization into trimethylsilyl ethers. The reproducibility of the combined trimethylsilylation and analysis of levoglucosan was about 2% for standard solutions, whereas the precision of the entire method for the sum of all three monosaccharide anhydrides (MAs) in real aerosol filter samples was about 5%. The method was applied to aerosol samples from urban and tropical locations. The atmospheric concentration of the MAs in fine (<2.5 microm) aerosols at a primary forest site in Rondjnia, Brazil, was on average 2.15 microg m(-3) during the dry season when intensive biomass burning occurs, which was almost 400 times higher than during the wet (nonburning) season. Urban total aerosols collected in Gent, Belgium, showed an average atmospheric concentration of MAs of 0.56 microg m(-3) for the winter season, which was a factor of 20 higher than for the summer season. The carbon in the MAs accounted on average for about 5.1% and 1.8% of the organic carbon in the Brazilian dry season and Gent winter aerosols, respectively. Levoglucosan was the major MA, with a relative abundance in the range of 76-93%. PMID- 11878394 TI - Charring characteristics of atmospheric organic particulate matter in thermal analysis. AB - The charring of organic materials during carbon analysis bythermal methods makes it difficult to differentiate elemental carbon (EC) from organic carbon (OC). Failure to correct for charring results in the overestimation of EC and the underestimation of OC. The charring characteristics andthermal behaviors of aerosol OC are studied by subjecting hexane and water extracts of ambient aerosols to various analysis conditions. The complete evolution of water-soluble organic carbon (WSOC) aerosol materials is found to require a temperature as high as 850 degrees C and the presence of oxygen. EC would be oxidized under these thermal conditions as well. As a result, thermal methods relying only on temperature for the differentiation of EC and OC would give unreliable OC and EC concentrations. Our investigation also reveals that WSOC accounts for a large fraction (13-66%) of charring, while hexane extractable organic compounds produce little charring. The extent of charring from WSOC, defined as the ratio between pyrolytically generated EC to the total WSOC, is found to increase with the WSOC loading in each analysis when the loadings are below a certain value. This ratio remains constant when the loadings are above this value. This may account for the high variability in the extent of charring among aerosol samples from different locations as well as among samples from a single location collected at different times. Charring is reduced if the residence time at each temperature step in a helium atmosphere is sufficiently long to allow for maximum C evolution at each step. Charring is also influenced by the presence of inorganic constituents such as ammonium bisulfate. For the few tested organic materials, it is observed that ammonium bisulfate enhances the charring of starch and cellulose but reduces the charring of levoglucosan. PMID- 11878395 TI - Catalyzed destructive adsorption of environmental toxins with nanocrystalline metal oxides. Fluoro-, chloro-, bromocarbons, sulfur, and organophosophorus compounds. AB - In the temperature range of 300-500 degrees C, solid nanocrystalline oxides react nearly stoichiometrically with numerous halocarbons, sulfur, and organophosphorus compounds. In some cases, the reaction efficiencies can be improved by the presence of a small amount of transition-metal oxide as catalyst; for example, Fe2O3 on CaO and mobile intermediate species such as FeCl3 or Fe(SO3)x are important in the catalytic process. Herein, a series of environmentally problematic compounds are discussed, including CCl4, COS, CS2, C2Cl4, CHCl3, CH2Cl2, CH3Cl, and (CH3O)2P(O)CH3. Nanocrystals of CaO coated with a thin layer of Fe2O3 (or other transition metals) =[Fe2O3]CaO, or intimately mixed =Fe2O3/CaO were compared with pure CaO. It was found that (a) the presence of a small amount of surface [Fe2O3] or other transition-metal oxide can have a marked effect on the destructive adsorption activity, (b) for some reagents, such as CCl4, C2Cl4, SO2 and others, the nanocrystalline CaO can react in stoichiometric amounts, especially if a transition-metal oxide catalyst is present, (c) although the reaction with dimethylmethylphosphonate is surface-limited, the nanocrystalline calcium oxide performed well and in high capacity, (d) nanocrystalline calcium oxide exhibits near stoichiometric activitywith several interesting sulfur containing compounds, such as COS and CS2, (e) unfortunately, most fluorocarbons were not destructively adsorbed at 500 degrees C under the conditions employed; however, some of these can be effectively mineralized over the calcium oxide at higher temperatures. These compounds include C2F6, C3F6, C2ClF3, and CHF3, and (f) upon reaction, surface areas decreased considerably, from about 100 to about 10 m2/g. The results of these experiments further demonstrate that, with the proper choice of catalytic material, some solid-gas reactions can be engineered to be rapid and essentially stoichiometric. PMID- 11878396 TI - Transport of silica colloids through unsaturated porous media: experimental results and model comparisons. AB - We present results on the migration of silica colloids through laboratory columns packed with partially saturated quartz sand. The transport of the silica colloids responds to changes in the steady-state volumetric moisture content (theta) and for low theta depends on the wetting history of the sand pack prior to colloid injection. A mathematical model that incorporates a first-order rate law to simulate film straining and a second-order rate law to simulate partitioning at air-water interfaces closely describes colloid transport and mass transfer over the range of experimental conditions tested. The mass-transfer parameters of the model are sensitive to changes in both the level of water saturation and the flow rate. A semiempirical expression, based on a modification of film-straining theory, accounts for the observed variation in the first-order rate coefficient with changes in theta and average porewater velocity. Our work indicates that the presence of the air phase substantially influences porewater concentrations of mineral colloids in water-unsaturated media and that the kinetics of particle removal attributed to air-water boundaries reflects the contribution of multiple mass-transfer mechanisms. PMID- 11878397 TI - Novel electrode system for electroflotation of wastewater. AB - Electroflotation (EF) is an attractive method in wastewater treatment. The heart of EF is the dimensionally stable oxygen evolution anode that is usually expensive. In this paper, we present a stable anode by coating IrOx-Sb2O5-SnO2 onto titanium. Accelerated life test showed that the electrochemical stability of the Ti/IrOx-Sb2O5-SnO2 anode containing only 2.5 mol % of IrOx nominally in the activated coating was even higher than that of the conventional Ti/IrOx anode. Its service life for electroflotation application is predicted to be about 20 yr. Voltammetric investigation demonstrated that the Ti/IrOx-Sb2O5-SnO2 anode could provide fast electron transfer. Moreover, the present anode was designed to be fork-like and arranged interlockingly at the same level as the cathode with a similar shape. Such an innovation in electrode configuration and arrangement allows bubbles produced at both electrodes to be dispersed into wastewater flow quickly and, therefore, enhances the effective contact between bubbles and particles, favorable for high flotation efficiency. In addition, the novel electrode system reduces the interelectrode gap to 2 mm, a spacing that is technically difficult for a conventional electrode system. This small gap results in a significant energy saving. Easy maintenance is found to be another advantage of this novel electrode system. PMID- 11878399 TI - Supercritical fluid extraction: an innovative tool for a fly ash-like model support. AB - The performance of the supercritical fluid extraction (SFE) technique to obtain a new and more appropriate model support for PCDD/F formation studies was investigated. To characterize fly ash and model supports and relate their chemical-physical properties, surface area and pore size were determined. To evaluate the influence on reactivity of the different model supports with respect to raw fly ash, a kinetic study of the thermal behavior of dibenzofuran (DF) was performed. Rate constants as well as the activation and thermodynamic parameters for the different systems were also compared. The model support obtained from SFE was very similar to raw fly ash from the structural, physical-chemical, and kinetic points of view. PMID- 11878398 TI - Removal and sequestration of iodide using silver-impregnated activated carbon. AB - Two silver-impregnated activated carbons (SIACs) (0.05 and 1.05 wt % silver) and their virgin (i.e., unimpregnated) granular activated carbon (GAC) precursors were investigated for their ability to remove and sequester iodide from aqueous solutions in a series of batch sorption and leaching experiments. Silver content, total iodide concentration, and pH were the factors controlling the removal mechanisms of iodide. Iodide uptake increased with decreasing pH for both SIACs and their virgin GACs. The 0.05% SIAC behaved similarly to its virgin GAC in all experimental conditions because of its low silver content. At pH values of 7 and 8 there was a marked increased in iodide removal for the 1.05% SIAC over that of its virgin GAC, while their performances were similar at a pH of 5. Scanning electron microscopy (SEM) and energy-dispersive X-ray (EDX) analyses prior to reaction with iodide showed the presence of metallic silver agglomerates on the 1.05% SIAC surface. After the reaction, elemental mapping with EDX showed the formation of silver iodide agglomerates. Oxidation of metallic silver was observed in the presence of oxygen, and the carbon surface appears to catalyze this reaction. When the molar ratio of silver to iodide was greater than 1 (i.e., M(Ag,SIAC) > M(I,TOTAL)), precipitation of silver iodide was the dominant removal mechanism. However, unreacted silver leached into solution with decreasing pH while iodide leaching did not occur. When M(Ag,SIAC) < M(I,TOTAL), silver iodide precipitation occurred until all available silver had reacted, and additional iodide was removed from solution by pH-dependent adsorption to the GAC. Under this condition, silver leaching did not occur while iodide leaching increased with increasing pH. PMID- 11878400 TI - Polynuclear aromatic hydrocarbon and particulate emissions from two-stage combustion of polystyrene: the effects of the secondary furnace (afterburner) temperature and soot filtration. AB - Laboratory experiments were conducted in a two-stage horizontal muffle furnace in order to monitor emissions from batch combustion of polystyrene (PS) and identify conditions that minimize them. PS is a dominant component of municipal and hospital waste streams. Bench-scale combustion of small samples (0.5 g) of shredded styrofoam cups was conducted in air, using an electrically heated horizontal muffle furnace, kept at Tgas = 1000 degrees C. Upon devolatilization, combustion of the polymer took place in a diffusion flame over the sample. The gaseous combustion products were mixed with additional air in a venturi and were channeled to a secondary muffle furnace (afterburner) kept at Tgas = 900-1100 degrees C; residence time therein varied between 0.6 and 0.8 s. At the exits of the primary and the secondary furnace the emissions of CO, CO2, O2, NOx, particulates as well as volatile and semivolatile hydrocarbons, such as polycyclic aromatic hydrocarbons (PAH), were monitored. Online analyzers, gravimetric techniques, and gas chromatography coupled to mass spectrometry (GC MS) were used. Experiments were also conducted with a high-temperature barrier filter, placed just before the exit of the primary furnace to prevent the particulates from entering into the secondary furnace. Results demonstrated the beneficial effect of the afterburner in reducing PAH concentrations, including those of mutagenic species such as benzo[a]pyrene. Concentrations of individual PAH exhibited a pronounced after burner temperature dependence, typically ranging from a small decrease at 900 degrees C to a larger degree of consumption at 1100 degrees C. Consumption of PAH was observed to be the dominant feature at 900 degrees C, while significant quantities of benzene and some of its derivatives, captured by means of carbosieve/Carbotrap adsorbents, were formed in the afterburner at a temperature of 1000 degrees C. In the primary furnace, about 30% of the mass of the initial polystyrene was converted into soot, while the total mass of PAH represented about 3% of the initial mass of combustible. The afterburner reduced the particulate (soot) emissions by only 20-30%, which indicates that once soot is formed its destruction is rather difficult because its oxidation kinetics are slow undertypical furnace conditions. Moreover, increasing the afterburnertemperature resulted in an increasing trend of soot emissions therefrom, which might indicate competition between soot oxidation and formation, with some additional formation occurring at the higher temperatures. Contrary to the limited effect of the afterburner, high-temperature filtration of the combustion effluent prior to the exit of the primary furnace allowed for effective soot oxidation inside of the ceramic filter. Filtration drastically reduced soot emissions, by more than 90%. Limited soot formation in the afterburner was again observed with increasing temperatures. The yields of both CO and CO2 were largely unaffected by the temperature of the afterburner but increased at the presence of the filter indicating oxidation therein. A previously developed kinetic model was used to identify major chemical reaction pathways involving PAH in the afterburner. The experimental data at the exit of the primary furnace was used as input to these model computations. A first evaluation of the predictive capability of the model was conducted for the case with ceramic filter and a temperature of 900 degrees C. The afterburner was approximated as a plug-flow reactor, and model predictions at a residence time of 0.8 s were compared to experimental data collected at its exit. In agreement with the experimental PAH concentration, only a minor impact of the afterburner treatment was observed for most species at 900 degrees C. OH was deduced to be the major reactant with a mole fraction about 4 orders of magnitudes higher than that of hydrogen radicals. Evidence for the need of further work on the quantitative assessment of oxidation of PAH and their radicals is given. PMID- 11878401 TI - A new, entropy based method to support waste and resource management decisions. AB - A new method is presented to quantify the potential of a system to concentrate or dilute substances. The approach is based on a comprehensive material flow analysis and Shannon's statistical entropy function that is transformed by a three-step procedure. The result is a new function that can be applied to any defined system with known mass-flows and substance concentrations. In combination with materials balances, the method yields quantitatively the Relative Statistical Entropy (RSE) and the Substance Concentrating Efficiency (SCE) of a given system. When applied to compare five solid waste incinerators of increasing technology levels, the method (1) gave evidence of the potential of modern incinerators to control material flows and (2) pointed to future directions for further improvement of incineration technology. Case studies demonstrate that the new tool is well suited to support and complement existing methods for decision making in waste and resource management such as LCA. PMID- 11878402 TI - Comment on "A kinetic study of the oxidation of S(IV) in seawater". PMID- 11878403 TI - Combination of protease inhibitors for the treatment of HIV-1-infected patients: a review of pharmacokinetics and clinical experience. AB - The use of highly active antiretroviral therapy, the combination of at least three different antiretroviral drugs for the treatment of HIV-1 infection, has greatly improved the prognosis for HIV-1-infected patients. The efficacy of a combination of a protease inhibitor (PI) plus two nucleoside analogue reverse transcriptase inhibitors has been well established over a period of up to 3 years. However, virological treatment failure has been reported in 40-60% of unselected patients within 1 year after initiation of a PI-containing regimen. This observation may, at least in part, be attributed to the poor pharmacokinetic characteristics of the PIs. Given as a single agent the PIs have several pharmacokinetic limitations; relatively short plasma-elimination half-lives and a modest and variable oral bioavailability, which is, for some of the PIs, influenced by food. To overcome these suboptimal pharmacokinetics, high doses (requiring large numbers of pills) must be ingested, often with food restrictions, which complicates patient adherence to the prescribed regimen. Positive drug-drug interactions increase the exposure to the PIs, allowing administration of lower doses at reduced dosing frequencies with less dietary restrictions. In addition to increasing the potency of an antiretroviral regimen, combinations of PIs may enhance patient adherence, both of which will contribute to a more durable suppression of viral replication. The favourable pharmacokinetics of PIs in combination are a result of interactions through cytochrome P450 3A4 (CYP3A4) isoenzymes and, possibly, the multi-drug transporting P-glycoprotein (P-gp). Antiretroviral synergy between PIs and non overlapping primary resistance patterns in the HIV-1 protease genome may further enhance the antiretroviral potency and durability of combinations of PIs. Many combinations contain ritonavir because this PI has the most pronounced inhibiting effects on CYP3A4. The combination of saquinavir and ritonavir, both in a dose of 400 mg twice-a-day, is the most studied double PI combination, with clinical experience extending over 3 years. Combination of a PI with a low dose of ritonavir (< or = 400 mg/day), only to boost its pharmacokinetic properties, seems an attractive option for patients who cannot tolerate higher doses of ritonavir. A recently introduced PI, lopinavir, has been co-formulated with low dose ritonavir, which allows for a convenient three-capsules, twice-a-day dosing regimen. In an attempt to prolong suppression of viral replication combinations of PIs are becoming increasingly popular. However, further clinical studies are needed to identify the optimal combinations for treatment of antiretroviral naive and experienced HIV-1-infected patients. This review covers combinations of saquinavir, indinavir, nelfinavir, amprenavir and lopinavir with different doses of ritonavir, as well as the combinations of saquinavir and indinavir with nelfinavir. PMID- 11878404 TI - HIV-1 reverse transcriptase mutations found in a drug-experienced patient confer reduced susceptibility to multiple nucleoside reverse transcriptase inhibitors. AB - HIV-1 reverse transcriptase (RT) genotypes were obtained from 13 patients treated with stavudine. No previously-reported mutations indicative of stavudine resistance were found in these patients and no novel mutations occurred in more than two patients. One patient, treated with stavudine for 1 month and treated previously with zidovudine, zalcitabine and lamivudine, carried a mutation at codon 75 of the RT (V75M). A chimeric virus, including the patient's RT sequence from codon 25 to codon 220, which carried the resistance mutations M41 L, D67N, T69D, K70R, L210W and T215Y in addition to V75M, displayed reduced susceptibility to multiple nucleoside RT inhibitors (NRTIs). Removal of V75M from this RT background resulted in a return of susceptibility to didanosine and lamivudine. Our data are in agreement with previous studies demonstrating the rarity of stavudine resistance mutations in stavudine-treated patients. However, we describe a new set of mutations, found in the RT of a heavily-treated patient, that can confer reduced susceptibility to multiple NRTIs. These results underscore the importance of increased vigilance for possible multiple-drug resistance in patients who have been heavily treated with NRTIs. PMID- 11878405 TI - Genotypic correlates of resistance to HIV-1 protease inhibitors on longitudinal data: the role of secondary mutations. AB - Direct sequencing of the pol gene was assessed retrospectively with protease inhibitor susceptibility in a longitudinal study. A total of 134 samples from 26 patients were analysed at regular intervals up to 2 years. Patients were included in virological failure despite indinavir, ritonavir or saquinavir based triple drug therapy. Both the type and number of certain secondary protease mutations modulated the effect of primary mutations on phenotypic resistance. This was notably applicable to L101/V, and to lesser extents to A711V/T. However, combinations of primary mutations, including 154V could predict resistance to the drug used and nelfinavir in more than 80%. In contrast, in vitro cross-resistance to amprenavir was rarely encountered. In addition, there was a relationship between a higher number of key mutations and poorer virological and clinical outcomes, respectively, from 6 and 3 months on. The key mutations were the protease mutations independently conferring phenotypic resistance and/or the reverse transcriptase mutations predicting treatment outcome. This relationship was independent from drug history, viral load and CD4 cell count measurements. In summary, even on a small sample size, sequence-based genotyping seems to be a good prognostic marker when performed longitudinally. In the context of primary resistance mutations, including additional secondary mutations, it may be useful in the prediction of phenotypic and clinical resistance. This should be assessed to optimize treatment monitoring before emergence of broadly cross-resistant virus. PMID- 11878406 TI - Virological and immunological responses to a once-a-day antiretroviral regimen with didanosine, lamivudine and efavirenz. AB - Current antiretroviral drugs cannot eradicate HIV infections, and persons living with HIV are often faced with very demanding daily therapeutic schedules that can induce poor adherence. More conveniently dosed and patient-friendly regimens are needed. We investigated, in this 48-week pilot study, a once-a-day highly active antiretroviral therapy regimen of didanosine, lamivudine and efavirenz. Seventy five consecutive antiretroviral-naive subjects were enrolled. Over the 48-week period, plasma HIV-RNA levels declined sharply, with a median decrease at the end of the observation time >3.4 log copies/ml. The proportion of patients achieving a plasma HIV-RNA level below the limit of detection (50 copies/ml) was 77% (intention to treat analysis) at the end of the study period. The mean CD4 cell count increased steadily over time from 251 to 459 cells/microl. Antiviral efficacy was similar in patients with a baseline HIV-RNA level above or below 100,000 copies/ml. However, patients with a baseline CD4 cell count <200 cells/microl showed a significantly worse virological response than that observed in patients with higher baseline CD4 counts. Overall 15 patients interrupted therapy. In four cases treatment interruption was due to lack of treatment response; three additional patients were lost to follow-up or withdrew informed consent. Eight patients stopped therapy because of adverse events. The once-daily combination of didanosine, lamivudine and efavirenz resulted in sustained viral suppression and was well-accepted by patients. This regimen may offer advantages in selected difficult-to-treat populations, allows directly observed therapy and can be a safe and effective alternative in antiretroviral-naive patients. These encouraging pilot results need to be confirmed in a comparative clinical trial. PMID- 11878407 TI - Ritonavir/saquinavir plus one nucleoside reverse transcriptase inhibitor (NRTI) versus indinavir plus two NRTIs in protease inhibitor-naive HIV-1-infected adults (IRIS study). AB - OBJECTIVES: To compare the efficacy, tolerability and safety of a ritonavir 400 mg/saquinavir hard gel fomulation 400 mg twice daily versus an indinavir 800 mg once every 8 h containing first-line protease inhibitor (PI) treatment regimen. METHODS: Open, randomized, multicentre clinical trial. PI-naive patients received either ritonavir/saquinavir and one nucleoside reverse transcriptase inhibitor (NRTI) or indinavir and two NRTIs. Intention-to-treat (ITT) and on-treatment (OT) analyses were performed. RESULTS: The baseline characteristics of the study participants were similar in both arms, 67 patients (37%) were naive to antiretroviral treatment. The proportion of patients who achieved a plasma viral load below the level of detection of 400 copies/ml at week 48 was 43% (39/90) in the ritonavir/saquinavir arm and 63% (57/90) in the indinavir arm (P=0.005, I PMID- 11878408 TI - Antiretroviral treatment failure and HIV-1 genotypic resistance in Sao Paulo, Brazil. PMID- 11878409 TI - The quality assurance programme for the national radioactivity surveillance network in Italy. AB - The quality assurance programme (QAP) carried out by the ENEA and ANPA for the laboratories belonging to the radioactivity surveillance network in Italy is outlined. The organisation of the QAP and the measurement campaigns performed from 1983 to 2000 are reviewed under different aspects such as network evolution, measurement types, standard sources needed for the calibration and intercomparison campaigns. network homogeneity and accuracy level, and main sources of systematic errors. The results show the effectiveness of the programme in obtaining a uniform accuracy level among the participating laboratories. Nevertheless a few sources of systematic errors are still present. PMID- 11878410 TI - Levels of depleted uranium in Kosovo soils. AB - The United Nations Environment Programme (UNEP) has performed a field survey at 11 sites located in Kosovo, where depleted uranium (DU) ammunitions were used by the North Atlantic Treaty Organization (NATO) during the last Balkans conflict (1999). Soil sampling was performed to assess the spread of DU ground contamination around and within the NATO target sites and the migration of DU along the soil profile. The 234U/238U and 235U/238U activity concentration ratios have been used as an indicator of natural against anthropogenic sources of uranium. The results show that levels of 238U activity concentrations in soils above 100 Bq x kg(-1) can be considered a 'tracer' of the presence of DU in soils. The results also indicate that detectable ground surface contamination by DU is limited to areas within a few metres from localised points of concentrated contamination caused by penetrator impacts. Vertical distribution of DU along the soil profile is measurable up to a depth of 10-20 cm. This latter aspect is of particular relevance for the potential risk of future contamination of groundwater. PMID- 11878411 TI - Validation of a geographic information system for the evaluation of the soil radon exhalation potential in South-Tyrol and Veneto, Italy. AB - The PERS (soil radon exhalation potential) project was promoted by ANPA (Italian Environmental Protection Agency) together with the Universita Cattolica del Sacro Cuore of Rome: the aim was to produce a geographic information system allowing the discovery of regions with different radon exhalation potential starting from some territorial knowledge. Some environmental measurements were carried out within this project in selected areas in South-Tyrol and Veneto. The measurement of radon in springwater and groundwater as well as in soil gas plays a decisive role for the validation of the algorithm for computing the PERS. Along with technical aspects, a possible use of the PERS method by the Regional Environmental Protection Agencies and by other agencies is discussed with the scope of identifying radon prone areas, as stated in the Italian 'Decreto Legislativo' 26 May 2000, n. 241. Moreover the forecasting power of PERS regarding indoor radon concentration is analysed. PMID- 11878412 TI - Determination of uranium isotopes in food and environmental samples by different techniques: a comparison. AB - The uranium concentration in 59 samples of bottled and tap water, mainly from northern Italy, was measured by different techniques. Results obtained by inductively coupled plasma mass spectrometry (ICP-MS), semiconductor alpha spectrometry and low level liquid scintillation counting with alpha/beta discrimination (LSC) have been compared. High resolution gamma spectrometry and semiconductor alpha spectrometry have been used to analyse uranium in a variety of organic and inorganic samples. Isotopic secular equilibrium in the 238U series may be lacking or hidden by auto-absorption phenomena, so caution should be used in evaluating gamma spectrometry data. Alpha spectrometry has also been used to ascertain the possible pollution from depleted uranium in the environment. PMID- 11878413 TI - Accuracy of field spectrometry in estimating 137Cs contamination in high altitude alpine soils. AB - In situ gamma ray spectrometry is an attractive method for providing information on the concentrations of radionuclides in the soil. This method requires separate knowledge of the soil density and of the radioactivity distribution relative to soil depth. In-field gamma spectrometry measurements were performed together with sampling of the soil at the same site for subsequent gamma spectrometry analysis in the laboratory. Results of 137Cs concentration were compared in order to: (i) verify the effectiveness of in situ gamma spectrometry in averaging local inhomogeneties in the soil; and (ii) quantify the sensitivity of the gamma ray spectrometry soil radioactivity evaluation with respect to the variability of the depth profiles obtained from a single site sampling. The site of study is a high altitude pasture, in the Alpine environment. PMID- 11878414 TI - Comparison of different methods for the assessment of the environmental gamma dose. AB - Four different techniques for the assessment of environmental gamma dose are discussed and compared: high pressure ionisation chamber (Reuter-Stokes type), TLD dosemeters (GR-200), in situ gamma ray spectrometry and dose evaluation from the activity concentration of radionuclides in the soil. Soil samples gathered from a field near Turin (North-West Italy) have been analysed with a p-type HPGe, and their radionuclide concentrations have been used to evaluate the dose at I m above the soil surface. At the same location the dose rate was evaluated with the in situ gamma ray spectrometry (with a portable n-type HPGe) and with the ionisation chamber and the TLD dosemeters. The contribution of cosmic rays was added to the dose rate calculated from the soil samples and the in situ gamma spectrometry. The results obtained with these four techniques agree well within 20%. PMID- 11878415 TI - Variations of the isotopic ratios of uranium in environmental samples containing traces of depleted uranium: theoretical and experimental aspects. AB - The possibility of using conventional analysis, such as gamma spectrometry and alpha spectrometry, for the detection of traces of depleted uranium (DU) in environmental samples has been investigated. The expected values have been compared with the experimental results obtained by using mollusc samples gathered in the Adriatic Sea. The analysis has shown that it is possible to detect DU. if the percentage composition is about 20% depleted uranium and 80% natural uranium, for a sample containing 10 Bq x kg(-1) of 238U. The possibility of extending this approach to samples with any given uranium concentration is investigated. PMID- 11878416 TI - Radon survey in schools in north-east Italy. AB - During the year 2000 the Regional Agency for Environmental Protection (ARPA) of Friuli Venezia Giulia together with the Pordenone Province carried out a survey to determine the radon concentration in the schools. About 900 measurements have been carried out in 300 schools located both in the mountainous and in the flat territory. Moreover, geological information was gathered. both on a regional scale and, where possible, on a small detailed scale. The purpose of this study is to consider the possibility of using all the data collected to discover some radon prone areas. The first results of this study seem to locate some radon prone areas where the cover consists of very permeable gravelly deposits. PMID- 11878417 TI - Experimental and measurement issues in natural radioactivity. AB - The paper deals with some open experimental problems in the field of natural radioactivity. Its aim is to stimulate discussion on the type of research efforts needed in the field and to be planned for the future. The review, which makes no claims to completeness. analyses some issues in gamma spectrometry of natural radionuclides, and the measurement of natural radioactivity in non-nuclear industries and of 222Rn and 220Rn concentration in air. PMID- 11878418 TI - In progress identification of radon prone areas: Toscana and Veneto. AB - Two studies aimed at defining radon prone areas in Toscana and Veneto, based on indoor measurements, are merged in this paper to extract the key points for a possible general methodology. In territorial units, the geometric mean (GM) and standard deviation (GSD) are evaluated from empirical data and the fraction of dwellings above the reference level (RL) is derived according to a log-normal model. In Toscana the sampling base is made of nearly 1,000 measurements (dwellings and kindergartens), apportioned according to the lithological classes (22) of the region. In Veneto 1,230 measurements in dwellings of the upper part of the region (estimates are given for the south), have been assigned to 6 x 5 km2 sequential units of the territory: the detection floor has been considered and results are referred to the regional mean dwellings condition with respect to the floor. Empirical GM and GSD values are corrected to increase statistical power based on two different methods. Maps of the percentages of dwellings with more than 200 Bq x m(-3) are presented. Areas with magmatic rocks in southern Toscana and northern parts of Vicenza and Belluno provinces in Veneto clearly emerge with more than 10% of dwellings above 200 Bq x m(-3). Efforts are required to establish proper confidence limits for estimates and to trace radon maps upon an administrative basis. PMID- 11878420 TI - Electric and magnetic fields generated by ac power lines: an application of advanced modelling tools in order to predict exposure levels. AB - In view of expected legislation changes imposing more severe limits on magnetic flux density, proposals are made based on theory and measurement for adjustment of Fower placement with regard to efficiency, cost and cost-benefit. PMID- 11878419 TI - Population exposure to electromagnetic fields generated by radio base stations: evaluation of the urban background by using provisional model and instrumental measurements. AB - Electromagnetic radiation, which is used by broadcasting and mobile telephone systems to transmit information, permeates the city environment. In order to properly evaluate population exposure to electromagnetic fields, knowledge of their intensity and spectral components is necessary. In this study the results of radiofrequency field monitoring carried out in Torino, a large town located in the north-west of Italy are shown: the variation of the electromagnetic field strength is evaluated as a function of the height from the ground, the location in the urban area and the frequency. separating the contributions of the different sources (broadcasting antennas and radio base stations for mobile phones). Furthermore, the contribution of the radio base stations is theoretically evaluated, adding the emissions off all installations situated in Torino and examining the field strength maps calculated, considering the orography, for different heights. The theoretical values are also compared with those measured in the frequency range of mobile telephony emissions. PMID- 11878421 TI - Calibration and evaluation of uncertainty in the measurement of environmental electromagnetic fields. AB - After a brief description of the reference systems for the calibration of the electromagnetic field meters set up at IEN and the Ivrea Department of ARPA, the procedures for the evaluation of the measurement uncertainty are discussed, with reference to the problem of identifying and quantifying the different uncertainty components. which are due both to the field meter characteristics and to the measurement procedure and conditions. The problem of how to compare a measurement result, which consists of the measurement value and its associated uncertainty, with a limit fixed by the standards is also faced. PMID- 11878422 TI - Dosimetric techniques for the evaluation of the em power absorption induced by cellular phones. AB - International guidelines. ICNIRP, and proposed standards. IEEE-ANSI, define basic restrictions in terms of rate of power absorbed for mass unit (SAR) for localised exposure of electromagnetic fields such as in the near field region of cellular phones. Compliance tests of cellular phones consist in checking that emitted power from mobile telecommunication equipment (MTE) is below the reference levels and they are performed by assessing the absorbed power in specific head phantoms. The outcome of this work is the description of experimental procedures for evaluating SAR induced into the head by localised exposure. The set up instituted for compliance testing of MTE is described with the different components: dielectric simulators, electric field sensors. scanning system, remote control and data recording. Calibration procedures of E sensors and uncertainty evaluation of measures are presented. Finally the results of a comparison among a few European laboratories and of a test of some hand-set devices are shown. PMID- 11878423 TI - Definition and development of an automatic procedure for narrowband measurements. AB - Measurements in the real environment, i.e. with several different electromagnetic field (EMF) sources and scattering problems, require an accurate analysis of problems concerning narrowband measurements. The aim of the present work was the development of an automatic procedure for narrowband electric field measurements in open sites with multiple sources, in order to perform accurate and reproducible measurements. Results regarding measurements carried out in a suburban site are reported. PMID- 11878424 TI - Measurement techniques for UMTS signals radiated by radio base stations. AB - In the most European countries radio coverage for the third radio mobile generation. i.e, the UMTS (Universal Mobile Telecommunications System), will soon be started. In the past few years, national laws specifying limits on exposure to electromagnetic fields have drawn much attention on electromagnetic test bed and measurement procedures for radio mobile equipment/systems. An overview is given of the UMTS system, showing the main characteristics of the radio access network UTRAN (UMTS Terrestrial Radio Access Network). An analysis is also provided as to the measurement techniques and related instrumentation for the electric field intensity radiated by a UMTS radio station. PMID- 11878425 TI - Assessment of the temporal trend of the exposure of people to electromagnetic fields produced by base stations for mobile telephones. AB - Monitoring of electric field levels produced by base stations (BSs) for mobile telephones of different typologies (TACS. GSM, DCS) has been carried out. Results show that BSs can be classified as 'business' or 'residential'. The mean value of six minutes averaged E-field value measured between 10 a.m. and 1 p.m. corresponds to the 84% of the maximum daily six minutes averaged value. Comparison between electromagnetic field levels produced by BSs and their phone traffic data, supplied by companies, is shown. Finally, on an hourly average basis, a daily curve has been constructed of the exposure trend produced by such installations. PMID- 11878426 TI - Analysis of the environmental factors for microcellular systems in urban areas. AB - In this study some general guidelines to be used for the installation of microcellular base stations in order to guarantee exposure levels have been identified. A detailed analysis of field strength levels around realistic urban microcellular antennae has been performed by means of experimental and theoretical evaluations as a function of antenna type and the characteristics of the surrounding environment. PMID- 11878427 TI - Electromagnetic field monitoring and control systems: state-of-the-art and work in-progress. AB - The development of EMF monitoring and control systems unquestionably represents one of the major innovations in the range of methodologies for evaluating, through measurement, the so-called environmental electromagnetic pollution. The interest recently shown by ARPA and municipal and provincial councils for setting up electromagnetic field monitoring and control systems has created significant developments in the field under examination. This paper attempts to provide a complete overview of the technical possibilities provided by electromagnetic field monitoring and control systems in the environment and their possible applications. Equipment currently available and under development is described, as well as the most significant work-in-progress in this sector. In the near future, several monitoring systems will be proposed and set up by municipalities and provinces, or directly by telecommunications companies. For this reason, it is more than ever necessary that agreement be reached at a national level to define a unified methodological and operating approach to enable monitoring data from different sources to be compared. PMID- 11878428 TI - Evaluation of long-term exposure to the magnetic field produced from power lines. AB - For some years, the Italian Regional Agencies for Environmental Protection (ARPA) have been engaged in the evaluation of long-term exposure to 50 Hz magnetic fields for receptors close to power lines. In this study, experiences and procedures are suggested in order to estimate long-term exposure levels in the case of magnetic induction produced contemporaneously by different power lines. In the simplest cases, the influence of a single line is evaluated by verifying the correlation factor between the variation of the current in each line and the measured magnetic induction. When the prevalence of a single line is not clear and systematic, much additional data (voltage, current, phases and geometrical configuration) have to be considered. The case of a school in Livomo near two lines. placed on the same supports, is described and a range of the most probable exposure values is given, corresponding to the phase change of the two current fluxes. PMID- 11878429 TI - Numerical and experimental analysis of e-field scattering from the ground for base station antennae. AB - Based on the numerical determination of the complete irradiation volume of a commercial RBS antenna--performed using the FDTD method and the Kirchhoff integral formula for near to far field transformation--open site estimations of the electric field are made and compared with experimentally measured values. To describe the actual behaviour of the radiation field, the inherently complex phasic nature of plane waves is taken into account, together with their two independent states of polarisation. This information is contained in the radiation pattern previously deduced. Moreover, a reflected contribution from flat ground is introduced, along with the line-of-sight ray. Amplitude and phase of the reflected wave are calculated using Fresnel formulae for stratified ground and two polarisation states, i.e. normal and parallel to the plane of incidence. Good agreement with measured values is achieved only by using such assumptions. PMID- 11878430 TI - Parallel FD-TD simulation of radiobase antennae. AB - The rigorous characterisation of the behaviour of a radiobase antenna for wireless communication systems is a hot topic both for antenna or communication system design and for radioprotection-hazard reasons. Such a characterisation deserves a numerical solution, and the use of a finite-difference time-domain (FD TD) approach is an attractive candidate. Unfortunately, it has strong memory and CPU-time requirements. Numerical complexity can be successfully afforded by using parallel computing. The parallel implementation of the FD-TD code, individuating the theoretical lower bound for its parallel execution time are discussed and the findings achieved on the APE/Quadrics massively parallel systems are presented. Results obtained from the simulation of actual radiobase antennae, clearly demonstrate that massively parallel processing is a viable approach to solving electromagnetic problems, allowing the simulation of radiating devices which could not be modelled through conventional computing systems. The tests showed a sustained computational speed equal to 17% of the theoretical maximum. PMID- 11878431 TI - Characterisation of spectrophotometers used for spectral solar ultraviolet radiation measurements. AB - Spectrophotometers used for spectral measurements of the solar ultraviolet radiation need to be well characterised to provide accurate and reliable data. Since the characterisation and calibration are usually performed in the laboratory under conditions very different from those encountered during solar measurements, it is essential to address all issues concerned with the representativity of the laboratory characterisation with respect to the solar measurements. These include among others the instrument stability, the instrument linearity, the instrument responsivity, the wavelength accuracy, the spectral resolution, stray light rejection and the instrument dependence on ambient temperature fluctuations. These instrument parameters need to be determined often enough so that the instrument changes only marginally in the period between successive characterisations and therefore provides reliable data for the intervening period. PMID- 11878432 TI - The calibration of personal dosemeters used for evaluating exposure to solar UV in the workplace. AB - In the framework of an epidemiological study regarding the correlation between solar UV radiation exposure and skin pathologies in a population of outdoor workers, the possibility of using polysulphone film personal dosemeters to quantify the Subjects UVB exposure has been evaluated. An original experimental set-up is presented. in a preliminary version, which ill be used both for solar irradiance spectroradiometric measurements and for the reading of personal dosemeters. The polysulphone absorption is similar to the CIE erythemal response curve. Due to UVB radiation exposure, the polysulphone film dosemeters photodegrade with a measrable absorbance change. The absorbance variation after the dosemeter exposure to UV radiation has been correlated to the UVB effective dose. The calibration curve obtained by this method may be particularly useful for the evaluation of small closes. The method will be used to quantify the personal exposure of workers whose exposure conditions are characterised by high variability. PMID- 11878433 TI - Effect of environmental factors on solar UV measurements. AB - For a correct understanding and interpretation of solar UV data sampled at ground level, several practical and theoretical problems of measurement are shown. In particular, the comparison between either different sites or instruments is thoroughly analysed, and the effects due to O3 level, albedo and cloudiness are discerned. The impact of clouds and their position with respect to the sun on the observation carried out both by spectral and broad-band instruments are also examined. PMID- 11878434 TI - Outdoor noise pollution: measurement of the main factors. AB - In industrial countries and particularly in Europe, a large part of the population considers noise pollution as one of the main environmental disturbances. After the publication of several national laws and regulations, noise predicting methods have been largely unproved. In some countries, the predicted acoustic levels have first to comply with the new limitations imposed by these regulations, and secondly to be experimentally verified in situ. In order to give some tools to the engineers, new measurement methods have been elaborated, and some of them standardised. These methods includes both the determination of outdoor sound levels as a function of the environmental conditions such as ground composition, presence of noise barrier and atmospheric effects, and the characterisation of the main parameters which can influence sound propagation, such as absorption coefficients of ground and materials covering noise barriers, and diffraction by various obstacles. PMID- 11878436 TI - EU enlargement and its potential impact on animal health. PMID- 11878435 TI - Measurement techniques for transport noise. AB - The noise front transport systems (roads, railways and aircraft) are increasing more and more both in space and in time and, therefore, they are still the major factor responsible for environmental noise pollution. The population exposed to transport noise is also increasing. and the corresponding health effects on people (i.e. annoyance and sleep disturbance) become more severe. Due to this current situation international and national legislation has been issued and implemented to reduce the harmful effects of such noise. This paper describes the techniques prescribed by recent Italian legislation to measure road, railway and aircraft noise. PMID- 11878437 TI - Retrospective analysis of serum and nasal mucus from cattle in Northern Ireland for evidence of infection with influenza A virus. AB - Eighty-four pairs of acute and convalescent serum samples collected in 1998 and 1999 from 17 outbreaks of respiratory disease, milk drop syndrome or diarrhoea in cattle were tested by haemagglutination inhibition against human influenza viruses A/Eng/333/80 (HIN1) and A/Eng/427/88 (H3N2). Antibodies to these viruses were present in the convalescent sera of 56.5 per cent and 58.8 per cent cattle tested, respectively, with 56 per cent of the animals seroconverting to one or both viruses. Titres were typically higher to A/Eng/427/88 (H3N2). Further testing of a subset of 21 of these serum pairs against the predominant H1N1 and H3N2 human and porcine strains circulating when the samples were collected revealed that the highest reactivity, in terms of both the magnitude of the recorded titres and the number of positive sera, was to human H3N2 strains. The titres to human H1N1 strains and to both porcine subtypes were low or absent. Attempts to isolate influenza A virus from nasal mucus or swab samples from 142 cattle from 46 cases of respiratory disease and/or milk drop syndrome by passage in embryonated specific pathogen-free eggs were unsuccessful. PMID- 11878438 TI - Diagnosis by ultrasonography of congestion of the caudal vena cava secondary to thrombosis in 12 cows. AB - This paper describes the clinical, ultrasonographic, radiographic and postmortem findings in 12 cows with thrombosis of the caudal vena cava. The principal clinical signs were chronic bronchopneumonia and fever in 11 cows; one cow had epistaxis and one cow bled from the mouth; eight cows had anaemia and leucocytosis, and the clotting time for the glutaraldehyde test was markedly decreased in all the cows; in nine of the cows the activity of gamma glutamyltransferase was high, suggesting chronic hepatic congestion. The most important ultrasonographic finding was congestion of the caudal vena cava attributable to thrombosis of the vein. In all the cows the caudal vena cava was round to oval on cross-section, rather than the normal triangular shape. The hepatic, splenic and portal veins were dilated in five, three and one cow, respectively. The results of radiography and endoscopy supported a diagnosis of bronchopneumonia, but there were radiographic changes in the diaphragmatic lung lobes that supported a diagnosis of vena caval disease in only four cows. Postmortem there was a thrombosis of the caudal vena cava in all the cows, and the thrombi were located in the thoracic, subphrenic and abdominal part of the caudal vena cava at the level of the liver in four, one and seven cows, respectively. In three cows, the thrombus was situated where a hepatic abscess had broken into the caudal vena cava, and in one cow it was at the site of a diaphragmatic abscess. In another cow, there was a fistula between the major bronchus of the right diaphragmatic lung lobe and the caudal vena cava where the thrombus was situated. Three cows had liver abscesses that had not broken into the caudal vena cava. There was severe bronchopneumonia in 11 of the cows, some of which also had multiple pulmonary abscesses. PMID- 11878439 TI - Incidence of Mycobacterium avium subspecies paratuberculosis in bulk-tank milk samples from different regions in Switzerland. PMID- 11878440 TI - First case of cat rabies in southern Brazil for 11 years. PMID- 11878441 TI - Prevalence of Brachyspira pilosicoli in Korean pigs, determined using a nested PCR. PMID- 11878442 TI - Tickborne encephalitis in a mouflon (Ovis ammon musimon). PMID- 11878443 TI - Duty of care regarding night visits. PMID- 11878444 TI - Access to the guide to professional conduct [letter]. PMID- 11878445 TI - 'Serum therapy' in pigs as a treatment for PMWS/PDNS. PMID- 11878446 TI - 'Serum therapy' in pigs as a treatment for PMWS/PDNS. PMID- 11878447 TI - Review of suspected adverse reactions surveillance scheme. PMID- 11878448 TI - Strategies for the control of avian influenza in Italy. PMID- 11878449 TI - Importation of the brown dog or kennel tick (Rhipicephalus sanguineus) into the UK. PMID- 11878451 TI - Changes in hip fracture epidemiology: redistribution between ages, genders and fracture types. AB - After several reports of increasing hip fracture incidence some studies have suggested a trend-break. In a previous study of hip fractures we forecast a 70% increase in the total number of fractures from 1985 up to year 2000. We therefore studied the incidence trend for the last 15 years and supply a new prognosis up to year 2010. We recorded all incident hip fractures treated in the county of Ostergotland, Sweden (approximately 400,000 inhabitants) 1982-96. A total of 11,517 hip fractures in men and women aged 50 years and above were included in the study after cross-validation between a computerized register of radiologic investigations and the hospital records. The projected number of fractures up to year 2010 was estimated by a Poisson regression model, considering both age and year of fracture in every single year 1982-96 for the respective fracture type and gender, and applied to the projected population. The annual number of hip fractures increased by 39% in men and 25% in women during the study period. Amongst men, the age-adjusted incidence of cervical fractures increased from 188 to 220/100,000 and of trochanteric fractures from 138 to 170/100,000. In women the incidence of cervical fractures decreased from 462/100,000 to 418/100,000 and of trochanteric fractures from 407/100,000 to 361/100,000. Cervical/trochanteric fracture incidence rate ratio leveled off, and also the female/male fracture rate ratio declined. A prognosis assuming that the incidence development will continue as during 1982-96, and a population in agreement with the forecast, predicts that the total age- and sex-adjusted number of hip fractures will decrease by 11% up to year 2010 compared with 1996. In women and men, however, a decrease of 19% and an increase of 7% respectively were projected. If the age- and sex-specific incidence remains at the same level as at the end of the study period, no significant change in the total numbers will occur. A trend-break was thus found in hip fracture incidence for women but not for men. Whether this is due to therapeutic and/or preventive measures in women is unknown. According to the most probable scenario a substantial increase in male trochanteric fractures (36%) is expected up to 2010, while all other hip fractures in both genders will decrease by 4-32% resulting in a total reduction of 11%. PMID- 11878450 TI - The use of placebo-controlled and non-inferiority trials for the evaluation of new drugs in the treatment of postmenopausal osteoporosis. AB - Registration of new agents for the treatment of postmenopausal osteoporosis has been based over the past few years on placebo-controlled phase III trials with the incidence of patients with new vertebral/nonvertebral fractures as the most usual primary endpoint. The use of a placebo in diseases where an active treatment is available has been a matter of debate following the update of the Declaration of Helsinki by the World Medical Association which questioned this trial design. Current regulatory recommendations within the European Union suggest that placebo-controlled trials are still the best option when assessing the efficacy and safety of new drugs intended for the treatment of postmenopausal osteoporosis. This suggestion seems to be in apparent contradiction with the current content of the Declaration of Helsinki. This paper addresses the ethics and feasibility of placebo-controlled trials in the treatment of postmenopausal osteoporosis, in the light of available therapeutic options, and discusses possible alternative approaches in those patients where placebo treatment could be deemed to be unethical. It is concluded that placebo-controlled trials remain the most efficient design to establish the efficacy and safety of a new agent for the treatment of postmenopausal osteoporosis. Such trials are feasible and ethically acceptable in patients with osteoporosis but without prevalent vertebral fractures. Conversely, in patients with prevalent vertebral fractures, placebo-controlled trials are ethically questionable and non-inferiority trials are more appropriate. A relative margin of non-inferiority of 20-30% is suggested, to be discussed on a case by case basis. PMID- 11878452 TI - Effect of collagen and mineral content on the high-frequency ultrasonic properties of human cancellous bone. AB - The technology surrounding ultrasonic bone assessment is evolving rapidly as investigators explore the utility of new ultrasonic parameters and different ultrasonic frequencies. This study had three main goals. The first was to perform in vitro measurements of the speed of sound (SOS) and normalized broadband ultrasonic attenuation (nBUA) in specimens of normal human cancellous bone using a 2.25 MHz broadband measurement system. The second was to explore the utility of a backscatter-based parameter called apparent integrated backscatter (AIB). The third goal was to investigate the roles that collagen and mineral content play in affecting each of these three ultrasonic parameters. This was accomplished by chemically treating the specimens to remove one or the other of these two important constituents of bone. Our results showed that in most cases SOS and nBUA correlated well (p < 0.05) with bone mineral density (BMD) as measured by quantitative computed tomography (QCT). In contrast, AIB did not correlate strongly with BMD. When the specimens were demineralized, decreases were produced in SOS (19-39%) and nBUA (44-58%). Changes produced in AIB were not significant except along the superoinferior direction, in which a 12% decrease was measured. When the specimens were decollagenized, decreases were produced in SOS (10-12%). In contrast, increases were produced in both nBUA (35-77%) and AIB (14-15%). From this study we conclude that high-frequency ultrasonic measurements may yield useful information about the content and organization of both collagen and mineral in cancellous bone. PMID- 11878453 TI - Computed tomography image analysis of the calcaneus in male osteoporosis. AB - The present study aimed to characterize bone microarchitecture assessed by computed tomography (CT) at the calcaneus in male subjects suffering from osteoporosis. Seventy-nine subjects were assessed (45 with osteoporosis and 34 control subjects matched for age). Osteoporosis was defined according to the World Health Organization classification either at the lumbar spine or at the femoral neck. Thirty-three subjects (73%) had a past history of low-energy fracture mainly represented by vertebral fractures (24/33). Nine axial sections (1 mm in width and 2 mm apart) were selected for each subject. Bone microarchitecture analysis was performed using structural (binary and skeletonized images but also skeletonization from gray levels) and fractal analyses. Bone densitometry by dual-energy X-ray absorptiometry (DXA) at the calcaneus was also performed in 73 cases. Bone mineral density (BMD) was decreased in osteoporotic patients compared with controls both at the lumbar spine and hip and also at the calcaneus (p<0.01). Also 14 microarchitectural features among 25 measured were significantly different between the two groups (p<0.01). The odds ratio for fracture per 1 control group standard deviation decrease were also significant for 13 structural features but also for BMD at the calcaneus. The odds ratios after adjustment for BMD at the calcaneus were significant for the following features (p<0.05): number of valleys, 2.8 (1.2 6.9); trabecular partition, 3.3 (1.3-7.9); apparent trabecular spacing, 1.8 (1.0 3.1); trabecular bone pattern factor, 2.2 (1.1-4.3); Euler number, 3.0 (1.1-8.7); node-to-terminus strut count, 3.3 (1.4-7.8); terminus-to-terminus strut count, 2.9 (1.2-6.9); and fractal dimension, 3.7 (1.5-9.7). Few and weak correlations were found between BMD at the calcaneus measured with DXA and features obtained from CT, suggesting that these two methods give different information about bone status. In conclusion, male osteoporosis is a disease characterized by decreased bone mass but also by microarchitectural deterioration of bone tissue which is partly independent of BMD. PMID- 11878454 TI - Recent decline in heavy outdoor work activity predicts occurrence of fractures among the home-dwelling elderly. AB - The contribution of reduced physical activity of a defined duration to the risk of fall-related fractures and serious soft tissue injuries is not known. We conducted a prospective population-based study among the home-dwelling elderly to examine the association between a recent decline in physical activity and the occurrence of fall-related fractures and soft tissue injuries. The study population consisted of representative sample of home-dwelling older adults who conducted heavy outdoor work activity at least once a week at baseline (n = 284; 136 men, 148 women) and among whom in 93 persons (33%) heavy outdoor work activity was found to have declined during the 2 1/2 years follow-up. Fall related fractures (n = 24) and serious soft tissue injuries (n = 49) were recorded from the time of the follow-up examination until the end of a further follow-up period lasting 3 1/2 years on average. A decline in heavy outdoor work activity did not predict the occurrence of soft tissue injuries (Mantel-Cox 0.795, p = 0.373), but a greater proportion of those with a decline (n = 14, 15%) than of others (n = 10, 5%) suffered fractures (Mantel-Cox 10.231, p = 0.001). Other risk factors for fractures were female sex (p = 0.03), slow choice reaction time (p = 0.02) and dependency as regards at least one basic activity of daily living (p = 0.01). According to the Cox proportional hazard model, the adjusted hazard ratio of fracture as regards a decline in heavy outdoor work activity was 2.7 (95% CI 1.14-6.62). A recent decline in heavy outdoor work activity predicts the occurrence of fractures, but not the occurrence of serious soft tissue injuries. Early recognition of a decline in physical activity may help in prevention of fractures among the elderly. PMID- 11878455 TI - German pediatric reference data for quantitative transverse transmission ultrasound of finger phalanges. AB - Quantitative ultrasound (QUS) of the finger phalanges is a useful tool in the assessment of disease- or age-related deterioration of bone. For studying the impact of juvenile diseases or growth disorders affecting the skeleton, a reference database for QUS parameters is needed. The aim of this study was to establish a calibrated reference database of parameters of transverse ultrasound transmission through juvenile finger phalanges. A total of 1328 children (650 females, 678 males; ages 3-17 years) were measured in Heidelberg and Kiel in order to establish a German reference database. Highly significant gender specific correlations (p<0.0001) were found between the QUS parameters amplitude dependent speed of sound (AD-SoS) and bone transmission time (BTT) versus age, body height and body mass index (BMI). For AD-SoS the correlation coefficients were R2 = 0.64 against age in males and R2 = 0.73 in females, R2 = 0.60 against body height in males and R2 = 0.68 in females, and R2 = 0.19 against BMI in males and R2 = 0.23 in females. For BTT the correlation coefficients were R2 = 0.74 against age in males and R2 = 0.79 in females, R2 = 0.75 against body height in males and R2 = 0.77 in females, and R2 = 0.32 against BMI in males and R2 = 0.35 in females. Age and height were the strongest determinants of QUS results. Gender specific differences were observed in AD-SoS (significant for ages 11-14 years and for 150-170 cm body height) and in BTT (significant for ages 7 and 11-17 years and for 160-170 cm body height). Tables of QUS parameters versus age and height can serve as a basis for the evaluation of the impact of skeletal diseases or growth disorders on phalangeal QUS. Depending on the type of disease or growth disorder, measurement results can be compared with age- or height- specific reference data. In this way a simple and radiation-free assessment of juvenile skeletal disorders using quantitative ultrasound might be possible in the future. PMID- 11878456 TI - In vivo assessment of architecture and micro-finite element analysis derived indices of mechanical properties of trabecular bone in the radius. AB - Measurement of microstructural parameters of trabecular bone noninvasively in vivo is possible with high-resolution magnetic resonance (MR) imaging. These measurements may prove useful in the determination of bone strength and fracture risk, but must be related to other measures of bone properties. In this study in vivo MR imaging was used to derive trabecular bone structure measures and combined with micro-finite element analysis (microFE) to determine the effects of trabecular bone microarchitecture on bone mechanical properties in the distal radius. The subjects were studied in two groups: (I) postmenopausal women with normal bone mineral density (BMD) (n = 22, mean age 58 +/- 7 years) and (II) postmenopausal women with spine or femur BMD -1 SD to -2.5 SD below young normal (n = 37, mean age 62 +/- 11 years). MR images of the distal radius were obtained at 1.5 T, and measures such as apparent trabecular bone volume fraction (App BV/TV), spacing, number and thickness (App TbSp, TbN, TbTh) were derived in regions of interest extending from the joint line to the radial shaft. The high resolution images were also used in a micro-finite element model to derive the directional Young's moduli (E1, E2 and E3), shear moduli (G12, G23 and G13) and anisotropy ratios such as E1/E3. BMD at the distal radius, lumbar spine and hip were assessed using dual-energy X-ray absorptiometry (DXA). Bone formation was assessed by serum osteocalcin and bone resorption by serum type I collagen C terminal telopeptide breakdown products (serum CTX) and urinary CTX biochemical markers. The trabecular architecture displayed considerable anisotropy. Measures of BMD such as the ultradistal radial BMD were lower in the osteopenic group (p<0.01). Biochemical markers between the two groups were comparable in value and showed no significant difference between the two groups. App BV/TV, TbTh and TbN were higher, and App TbSp lower, in the normal group than the osteopenic group. All three directional measures of elastic and shear moduli were lower in the osteopenic group compared with the normal group. Anisotropy of trabecular bone microarchitecture, as measured by the ratios of the mean intercept length (MIL) values (MIL1/MIL3, etc.), and the anisotropy in elastic modulus (E1/E3, etc.), were greater in the osteopenic group compared with the normal group. The correlations between the measures of architecture and moduli are higher than those between elastic moduli and BMD. Stepwise multiple regression analysis showed that while App BV/TV is highly correlated with the mechanical properties, additional structural measures do contribute to the improved prediction of the mechanical measures. This study demonstrates the feasibility and potential of using MR imaging with microFE modeling in vivo in the study of osteoporosis. PMID- 11878457 TI - The role of cytokines in the changes in bone turnover following bone marrow transplantation. AB - Osteoporosis is a common disease among patients undergoing transplantation and a loss of bone mass is usually detected after bone marrow transplantation (BMT), particularly during the immediate post-BMT period. Post-BMT bone loss is primarily related to gonadal dysfunction and immunosuppression. Cytokines, especially interleukin 6, play an important role in the pathogenesis of postmenopausal osteoporosis. However, the pathogenetic role of cytokines in post BMT bone loss is unknown and data on the changes of cytokines in accordance with bone turnover markers are scarce. The aim of this study was to assess the relationship between bone turnover markers and cytokines, which are regularly sampled at peripheral blood and bone marrow before and after allogeneic BMT. This prospective study included two analyses. The first was a study of 46 BMT recipients (M/F 28/18), examining the relationship between bone turnover markers and serum cytokines that were measured before and at 1 week, 2 weeks, 3 weeks, 4 weeks and 3 months after BMT. Serum intact parathyroid hormone was measured before BMT and at 3 weeks after BMT and its relation to other cytokines and bone turnover markers was evaluated. The second analysis was a study of 14 (M/F 9/5) of 46 patients in whom bone marrow plasma cytokines [interleukin 6 (IL-6) and tumor necrosis factor alpha (TNF-alpha)] were measured at 3 weeks after BMT. The relationship between bone marrow plasma cytokines and bone turnover markers was studied because bone marrow is the microenvironment where the real changes in bone turnover occur. Serum type I collagen carboxyterminal telopeptide (ICTP), a bone resorption marker, increased progressively until 4 weeks (peak) after BMT and then decreased thereafter. Serum osteocalcin, a bone formation marker, decreased progressively until 3 weeks after BMT and then increased thereafter. Serum IL-6 increased until 2 weeks after BMT and declined thereafter. Serum TNF alpha increased until 3 weeks after BMT and declined thereafter. There was a significant positive correlation between serum ICTP and bone marrow IL-6 levels at 3 weeks after BMT, when a marked change in bone metabolism occurs following BMT. However, a correlation between bone turnover markers and bone marrow TNF alpha or peripheral blood cytokines was not found. At 3 months after BMT, there was a significant negative correlation between the mean daily steroid dose and the serum osteocalcin level (r = -0.43, p < 0.05). The correlation between the Mean daily steroid dose and serum ICTP was also significant (r = 0.41, p < 0.05). Our data suggest that the progressive increase in bone resorption during the immediate post-BMT period is related to both steroid dose and the increase in bone marrow IL-6, which is a potent stimulator of bone resorption in vivo. PMID- 11878458 TI - Proximal femur geometry to detect and distinguish femoral neck fractures from trochanteric fractures in postmenopausal women. AB - Some proximal femur geometry (PFG) parameters, measured by dual-energy X-ray absorptiometry (DXA), have been reported to discriminate subjects with hip fracture. Relatively few studies have tested their ability to discriminate femoral neck fractures from those of the trochanter. To this end we performed a cross-sectional study in a population of 547 menopausal women over 69 years of age with femoral neck fractures (n = 88), trochanteric fractures (n = 93) or controls (n = 366). Hip axis length (HAL), neck-shaft angle (NSA), femoral neck diameter (FND) and femoral shaft diameter (FSD) were measured by DXA, as well as the bone mineral density (BMD) of the nonfractured hip at the femoral neck, trochanter and Ward's triangle. In fractured subjects, BMD was lower at each measurement site. HAL was longer and NSA wider in those with femoral neck fractures. With logistic regression the age-adjusted odds ratio (OR) for a 1 standard deviation (SD) decrease in BMD was significantly associated at each measurement site with femoral neck fracture (femoral neck BMD: OR 1.9, 95% confidence interval (95% CI): 1.4-2.5; trochanter BMD: OR 1.6, 95% CI 1.2-2.0; Ward's triangle BMD: OR 1.7, 95% CI 1.3-2.2) and trochanteric fracture (femoral neck BMD: OR 2.6, 95% CI 1.9-3.6; trochanter BMD: OR 3.0, 95% CI 2.2-4.1; Ward's triangle BMD: OR 1.8, 95% CI 1.4-2.3). Age-adjusted OR for 1 SD increases in NSA (OR 2.2, 95% CI 1.7-2.8) and HAL (OR 1.3, 95% CI 1.1-1.6) was significantly associated with the fracture risk only for femoral neck fracture. In the best predictive model the strongest predictors were site-matched BMD for both fracture types and NSA for neck fracture. Trochanteric BMD had the greatest area (0.78, standard error (SE) 0.02) under the receiver operating characteristic curve in trochanteric fractures, whereas for NSA (0.72, SE 0.03) this area was greatest in femoral neck fractures. These results confirm the association of BMD with proximal femur fracture and support the evidence that PFG plays a significant role only in neck fracture prediction, since NSA is the best predictive parameter among those tested. PMID- 11878459 TI - Biological measurement of estrogenic activity in urine and bile conjugates with the in vitro ER-CALUX reporter gene assay. AB - Although estrogens are excreted as biologically inactive conjugates, they can be reconverted to an active form, possibly by bacteria. A simple method was developed to deconjugate estrogen metabolites present in human urine and fish bile back to active estrogens by enzymatic hydrolysis with beta-glucuronidase or live Escherichia coli cells. Deconjugated extracts were tested for estrogenic activity in the in vitro stable estrogen receptor-mediated chemical-activated luciferase gene expression (ER-CALUX) assay. Estrogen glucuronides in urine obtained from human males and females were effectively converted to active forms after incubation with beta-glucuronidase or E. coli. The highest estrogenic activity was found in deconjugated metabolites from urine of a pregnant woman, in which levels up to 3,000 nmol estradiol equivalents per liter of urine were found after overnight incubation of urine with E. coli. Bile sampled from male bream and flounder from various freshwater and marine locations was also deconjugated and a good correlation was found between high biliary estrogenic activity and elevated levels of xenoestrogenic activity in surface water as well as in plasma vitellogenin. Therefore, the measurement of deconjugated bile could form a useful (indirect) biomarker for internal dose of xenoestrogens in male fish. PMID- 11878460 TI - Studies on the effect of soil aging on the toxicity of pyrene and phenanthrene to a soil-dwelling springtail. AB - Soil samples spiked with five concentrations of pyrene and phenanthrene were aged for 0, 10, 40, and 120 d before toxicity was investigated using a standardized bioassay with the soil-dwelling collembolan Folsomia fimetaria L. Toxic effects were measured as reductions in survival and reproductive output after 3 weeks of exposure. Both pyrene and phenanthrene were degraded in the test system during storage, phenanthrene to a higher degree than pyrene. However, when toxic effects of the tested substances were calculated on the basis of measured concentrations of the parent compounds, toxicity was unaffected by storage for up to 120 d. Many studies have shown a negative correlation between aging and the biodegradation of polycyclic aromatic hydrocarbons (PAHs), indicating a reduction in the bioavailability of PAHs to micro-organisms with time. Our results indicate that a reduction in toxicity, as related to the measured concentrations of the parent compounds in the soil, may not always be expected for aged soils. Provided that metabolites of pyrene and phenanthrene did not significantly contribute to the toxicity in aged soil samples, a possible explanation for the absence of aging effects is the high test concentrations used in combination with the low content of organic carbon (1.6%) in the test soil. PMID- 11878461 TI - The influence of redox potential on the degradation of halogenated methanes. AB - To determine the influence of redox potential on the reaction mechanism and to quantify kinetics of the dechlorination by digester sludge, the test compounds trichlorofluoromethane (CFCl3), carbon tetrachloride (CCl4), and chloroform (CHCl3) were incubated in the presence of sludge and variable concentrations of reducing agent. Different sources of dehalogenation were examined, including live sludge and heat-killed sludge, and abiotic mechanisms were quantified in the absence of sludge. Batch incubations were done under redox conditions ranging from +/-534 to -348 mV. The highest rates for the dehalogenation of the three compounds were observed at -348 mV. The dechlorination rate of all the compounds by the heat-resistant catalysts was approximately twofold higher than the live treatments. It was proposed that the higher degradation rates by heat-killed sludge were due to the absence of physical barriers such as cell wall and cell membranes. There was no abiotic dechlorination of CFCl3, whereas CCl4 and CHCl3 were both reduced in the absence of sludge catalyst by Ti (III) citrate at > or =2.5 mM. The degradation pathways of CFCl3 and CHCl3 appeared to be only partially reductive since the production of reduced metabolites was low in comparison with the total amount of original halogenated compounds degraded. For CFCl3, the partial reductive degradation implied that different intra- and extracellular pathways were concurrent. The Gibbs free energy and the redox potential for the dehalogenation reactions utilizing Ti (III) citrate and acetate as electron donors are reported here for the first time. PMID- 11878462 TI - Photodechlorination of 3,3'-dichlorobenzidine in water. AB - Laboratory experiments have been conducted to elucidate the photochemical behavior of 3,3'-dichlorobenzidine (DCB) and its congeners in aquatic systems. Photodechlorination of DCB was observed in aqueous samples that were irradiated with monochromatic radiation from a variable-wavelength laser at several wavelengths in the range 300 nm < or = lambda < or = 360 nm. Analytical measurements performed by high-performance liquid chromatography (HPLC) and Fourier-transform ion cyclotron resonance (FT-ICR) mass spectrometry revealed that 3-chlorobenzidine (MCB) was produced as a transient intermediate in the photodechlorination process, with subsequent photodechlorination to yield benzidine as a stable photoproduct. Data obtained from these measurements also suggest the existence of other (unidentified) reaction pathways and products. The time-course measurements of chlorobenzidine congener concentrations and irradiance history were combined with a simple reaction model to develop estimates of reaction kinetics. Model predictions were consistent with results obtained for all wavelengths investigated. PMID- 11878463 TI - Estrogenic activity measured in a sewage treatment works treating industrial inputs containing high concentrations of alkylphenolic compounds--a case study. AB - Chemical analyses were combined with a biological assay to investigate the main estrogenic chemicals as they passed through a sewage treatment works (STW) and entered a river. The STW studied was unusual in that it received wastewater from the textile trade. This wastewater was shown to contain high concentrations of alkylphenol polyethoxylates and their degradation products, such as nonylphenol. High-performance liquid chromatography fractionation, combined with biological assay, showed that the majority of the estrogenic activity was contributed by the alkylphenolic chemicals and the natural estrogens 17beta-estradiol and estrone. Despite removal of a high proportion of the alkylphenolic chemicals by the various treatment processes within the STW, concentrations in the final effluent were still high compared to most other STW effluents in the United Kingdom. The effluent was very estrogenic to caged fish, as was the river water 2 and 5 km downstream of the STW, even though less so. Using various approaches, attempts were made to determine which group of chemicals contributed most to the estrogenic activity of the effluent. The analysis suggested that, in this unusual situation, the alkylphenolic chemicals may contribute the majority of the estrogenic activity of the effluent. However, this conclusion was based on a number of uncertainties that are presently unresolved and hence can be considered only tentative. PMID- 11878464 TI - Reduction in the estrogenic activity of a treated sewage effluent discharge to an English river as a result of a decrease in the concentration of industrially derived surfactants. AB - As a result of the introduction of tighter discharge limits and effluent treatment processes at source, the concentration of alkylphenol ethoxylates and nonylphenol present in the final effluent discharge from a sewage treatment works that treats trade effluent from the textiles industry was reduced. The estrogenic effects of the final effluent discharge to the Aire River were compared over a four-year period during which various treatment measures were introduced. Male rainbow trout exposed to the effluent on four occasions in consecutive years (1994-1997) showed a reduction in the level of induced vitellogenesis between 1994 and 1997. A marked decrease in gonadosomatic index (GSI) and increase in heptaosomatic index (HSI) was measured in fish exposed to the effluent in 1994. In successive years, these differences diminished, and in the case of the GSI no measurable difference was observed between fish exposed to the final effluent or those in the control group in 1997. However, an increase in HSI was still measurable in 1997 in fish exposed to the final effluent and at sites farther downstream. The reduction in the effects of the effluent paralleled the reduction in the concentration of nonylphenol as well as its mono- and diethoxylates, which have been demonstrated to produce estrogenic effects in trout exposed to these compounds in the laboratory. This study demonstrates that the setting of more restricted discharge limits for known estrogenic chemicals of industrial origin can lead to significant reductions in the estrogenic activity of the watercourses into which the effluents are discharged. PMID- 11878465 TI - Impact of local agricultural and industrial practices on organic contamination of little egret (Egretta garzetta) eggs in the Rhone Delta, southern France. AB - The purpose of this study was to determine the importance of the effects of local agricultural and industrial activities on the contamination of little egret eggs laid in the Camargue area, a protected wetland area in the Mediterranean region. Despite the fact that human activity is widespread in the Camargue, concentrations of lindane (0.01-0.7 microg/g wet wt) and of polycyclic aromatic hydrocarbons (<0.5 microg/g wet wt) do not seem to be deleterious-at least for herons. Residues of dichlorodiethyltrichlorethylene (DDE) were usually below 1 microg/g wet weight. The principal threat comes from high concentrations of polychlorinated biphenyls (PCBs; 0.1-12 microg/g wet wt), which are known to have teratogenic effects in other bird species at similar concentrations and are thought to originate locally (sediments of the delta and industrial complex of Fos/Mer). For this reason, particular interest should be focused on the water/sediment quality of the Rhjne River (France), on industrial sewage, and on wildlife contamination. A significant but weak link between colony site and lindane concentration was detected in eggs and may be attributed to the fact that our egg collection from each sampled colony was not synchronized with pesticide spread. A stronger site effect was observed for PCBs. Eggs collected near the industrial complex of Fos/Mer (petroleum, power plants) displayed the highest median concentration and the most contaminated eggs. Such results warrant thorough monitoring of this colony. PMID- 11878466 TI - Response of the amphibian tadpole (Xenopus laevis) to atrazine during sexual differentiation of the testis. AB - Xenopus laevis tadpoles were exposed for 48 h during sexual differentiation to atrazine at 21 microg/L under static laboratory conditions at 21+/-0.5 degrees C. After this exposure period, tadpoles were fixed and the kidney-gonad complex was microdissected. Quantitative histological analysis of the gonad revealed a 57% reduction in testicular volume among atrazine-exposed tadpoles. In addition, primary spermatogonial cell nests that represent germ cells for the life of the organism were reduced by 70%. Nursing cells, which provide nutritive support for the developing germ cells, had declined by 74%. Testicular resorption was observed among 70% and aplasia or failure of full development of the testis was recorded in 10% of the atrazine-exposed tadpoles. Because cell nests represent the pool of primordial germ cells for the reproductive life of the organism, the combined reduction in spermatogonial cell nests and nursing cells suggest that a pulse exposure to 21 microg/L of atrazine during sexual differentiation could significantly reduce reproduction during the reproductive life of these animals. PMID- 11878467 TI - Biochemical biomarkers of pollution in the clam Chamaelea gallina from south Spanish littoral. AB - Fourteen biochemical pollution biomarkers were analyzed in the clam Chamaelea gallina sampled at seven South-Spanish littoral sites at different times in 1999. They included enzymes that regenerate reduced nicotinamide adenine dinucleotide phosphate (NADPH) or maintain the cytosol-reduced (glucose-6-P- and isocitrate dehydrogenases, glutathione reductase), that decompose reactive oxygen species (catalase, superoxide dismutase, glutathione peroxidase), or detoxify glutathione reactive electrophiles (soluble and microsomal glutathione transferases, glyoxalases II and I). The levels of reduced glutathione, malondialdehyde, and metallothionein and the glutathione redox status were also determined. Clams from Huelva sites with higher contaminant loads and metallothionein levels showed higher antioxidant and glutathione-related activities and a decreased glutathione level, lower malondialdehyde content, and a less oxidized glutathione status. This suggests that clams chronically exposed to contaminants released by Huelva and Guadalquivir Estuaries are better protected from oxidative stress than reference animals. Most biomarkers showed six-month cyclicity with up to threefold amplitudes, further supporting the inverse relationship between antioxidant defenses and oxidative damages. Thus the lower antioxidant defenses in June fit to maximum oxidative damages, whereas the high antioxidant defenses found in March and September through October agree with lower lipid damages and less oxidized glutathione status. The effect of environmental or endogenous factors on this cyclic response is discussed. PMID- 11878468 TI - Citrate-mediated increase in the uptake of weathered 2,2-bis(p-chlorophenyl) 1,1 dichloroethylene residues by plants. AB - Experiments were conducted to determine the ability of citrate to enhance the plant uptake of weathered 2,2-bis(p-chlorophenyl)1,1-dichloroethylene (p,p'-DDE) from soil. Plots containing three rows of clover, mustard, hairy vetch, or rye grass were constructed in soils containing p,p'-DDE. On 11 occasions, the rows of each crop received water or sodium citrate (0.005 or 0.05 M). For each crop, there were significant reductions in p,p'-DDE concentration in the soil fractions (near root and rhizosphere) closely associated with the plant versus bulk soil. The roots of each crop accumulated 2 to 5 times more of the weathered contaminant (dry wt) than present in the bulk soil. Citrate (0.05 M) increased the concentration of p,p'-DDE in the roots of clover, mustard, and hairy vetch by 39% compared with vegetation that received water. In batch desorption studies, the release of weathered p,p'-DDE was significantly greater in the presence of 0.05 M citrate than in water. Citrate increased the extracted aqueous concentrations of five metal ions (Al, Fe, Ca, K, Mn) from soil by five- to 23-fold over distilled water. We hypothesize that citrate physically disrupts the soil through chelation of structural metal ions and release of bound humic material, facilitating p,p' DDE availability and uptake by plants. PMID- 11878469 TI - Effects of handling on heat shock protein expression in rainbow trout (Oncorhynchus mykiss). AB - As part of an effort to validate the use of heat shock proteins (HSPs) as biomarkers of exposure to and effects of contaminants, we evaluated the effect of two handling regimens on the induction of HSP 60 and 70 in rainbow trout (Oncorhynchus mykiss). Fish were acclimated to laboratory conditions for several weeks before the beginning of the experiment. Fish were then captured by net, placed in a cooler for 1 h while being transported in a truck, returned to their original tanks, then sacrificed 6 to 8 h later. Tricaine methane sulfonate (MS 222) was used during different phases of handling to reduce handling stress. Heat stressed fish were included in the experiment as a positive control. Muscle, liver, gills, and heart were analyzed for HSP 60 and 70 by immunoblotting. We found no effect of any handling regimen on the induction of HSPs. These findings suggest that the capture and transport of fish for environmental monitoring purposes should not interfere with the use of stress proteins as biomarkers. PMID- 11878470 TI - Sublethal effects in avocet and stilt hatchlings from selenium-contaminated sites. AB - Excess selenium (Se) in the aquatic food chain is embryotoxic and teratogenic to avocets, stilts, and other waterbirds. American avocet (Recurvirostra americana) and black-necked stilt (Himantopus mexicanus) eggs were collected from three sites in the Tulare Lake Basin of California, USA, and hatched in the laboratory. These sites included the Tulare Lake Drainage District-north (TLDD-N, water 2.5 ppb Se), TLDD-south (TLDD-S, water 8.6 ppb Se), and Westfarmers (WF, water 190 ppb Se). Highest egg Se concentrations occurred at WF (geometric mean 31.4 ppm dry wt for avocets and 20.5 ppm dry wt for stilts). Mean egg Se concentrations were 6.7 ppm for avocets and 8.4 ppm for stilts at TLDD-S, and 3.3 ppm for avocets and 2.3 ppm for stilts at TLDD-N. Hatching success and incidence of malformations did not differ among sites, but yolk sac-free hatching weights and bone lengths were less for avocets at the WF site, whereas liver weights and liver to body weight ratios were greater at that site. With increasing Se concentration, oxidative stress was most apparent in avocet hatchlings from WF: hepatic glutathione (GSH) peroxidase activity increased, glucose-6-phosphate dehydrogenase activity decreased, and oxidized glutathione (GSSG) concentration as well as the ratio of GSSG to reduced GSH concentration increased. In stilts, hepatic GSH concentration was lower in WF hatchlings. In conclusion, our findings of Se-impaired embryo growth and hepatotoxicity in avocet hatchlings suggest that oxidative stress observed in hatchlings may be related to these biological effects and may serve as a potential bioindicator of subsequent impaired functions. PMID- 11878471 TI - Genetic effects of mercury contamination on aquatic snail populations: allozyme genotypes and DNA strand breakage. AB - Allozyme data and DNA strand break frequencies were compared among populations of Pleurocera canaliculatum from five sites with varying mercury contamination on the North Fork Holston River (NFHR) in southwestern Virginia, USA. Allozyme genotype frequencies for four loci were significantly different between populations from the three most highly contaminated sites and those from two lesser contaminated sites. In addition, heterozygosity at three of these loci was significantly lower in the populations from the most highly contaminated sites. The DNA strand break frequency was significantly correlated to whole-body total mercury concentration in snails from three sites. These data add to the evidence supporting the use of DNA strand breakage as an indicator of chemical contamination and the use of allozyme analysis as a marker of contamination and possible selection for pollution resistance. However, the relationship between contaminant-induced changes in the genetic variation of enzymes of central metabolism and the functionalities upon which selection for resistance may act remain unclear, and mechanisms other than selection for resistance must be considered. Use of enzymes from other biochemical pathways may be appropriate for other species or for those under other chemical pollution pressures. PMID- 11878472 TI - Environmentally relevant concentrations of ammonium perchlorate inhibit thyroid function and alter sex ratios in developing Xenopus laevis. AB - Embryos and larvae of the South African frog Xenopus laevis were exposed to ammonium perchlorate (AP) or control medium for 70 d. The dosage levels (59 ppb, 14,140 ppb) bracketed a range of perchlorate concentrations measured in surface waters at the Longhorn Army Ammunition Plant (LHAAP) in Karnack, Texas, USA. The experiment also included a 28-d nontreatment recovery period to assess the reversibility of AP effects. There were no significant effects of AP on mortality or hatching success. There were no effects of AP on developmental abnormalities such as bent/asymmetric tails or edema. Ammonium perchlorate inhibited forelimb emergence, the percentage of animals completing tail resorption, and hindlimb development during the 70-d exposure period. Only the upper AP concentration reduced whole-body thyroxine content, whereas both concentrations caused significant hypertrophy of the thyroid follicular epithelium. Both concentrations of AP caused a skewed sex ratio, significantly reducing the percentage of males at metamorphosis. The effects of AP on metamorphosis and thyroid function were reversed during the 28-d nontreatment recovery period. We conclude that AP inhibits thyroid activity and alters gonadal differentiation in developing X. laevis. These effects were observed at concentrations at or below concentrations reported in surface waters contaminated with ammonium perchlorate, suggesting that this contaminant may pose a threat to normal development and growth in natural amphibian populations. PMID- 11878473 TI - Effects of atrazine and cyanazine on chlorpyrifos toxicity in Chironomus tentans (Diptera: Chironomidae). AB - Toxicities of two triazine herbicides (atrazine and cyanazine) and an organophosphate insecticide (chlorpyrifos) were evaluated individually and with each herbicide in binary combination with chlorpyrifos using fourth-instar larvae of the aquatic midge, Chironomus tentans. Chlorpyrifos at 0.25 microg/L resulted in an effect in less than 10% of midges in 48-h acute toxicity bioassays. Neither atrazine nor cyanazine alone at relatively high concentrations (up to 1,000 microg/L) caused significant acute toxicity to C. tentans. However, atrazine and cyanazine caused significant synergistic effects on the toxicity of chlorpyrifos when midges were exposed to mixtures of atrazine or cyanazine (10, 100, 1,000 microg/L) with chlorpyrifos (0.25 microg/L). At fixed concentrations (200 microg/L) of the herbicides, toxicity of chlorpyrifos was enhanced by 1.8- and 2.2-fold by atrazine and cyanazine, respectively, at the 50% effective concentration levels. Although atrazine and cyanazine are not effective inhibitors of acetylcholinesterase (AChE) in vitro, the synergism of the two triazine herbicides with chlorpyrifos was associated with increased in vivo inhibition of AChE in midges. We observed a positive correlation between the degree of inhibition of AChE and the concentration of atrazine or cyanazine in the presence of a fixed concentration of chlorpyrifos. It is possible that these herbicides may affect cytochrome P450 enzymes to confer synergistic effects on the toxicity of chlorpyrifos. PMID- 11878474 TI - Isopod gut microflora parameters as endpoints in toxicity studies. AB - Terrestrial isopods Porcellio scaber (Crustacea) were fed for five weeks on food contaminated by 250, 500, or 1,000 microg of Cd/g or for 10 d on diets with 50 or 250 microg Cd/g food. In both experiments, fecal production rate and colony forming units (CFUs) in the guts were determined. In addition, at the end of 10 d, each distinct colony morphotype obtained in gut samples was purified and characterized. Isolates were separated into 25 groups based on morphological and biochemical characteristics. These bacterial groups were used as units for calculating Shannon equitability indices (J) for each gut. The relative frequencies of the 25 bacterial units were determined in both cadmium groups (50 or 250 microg Cd/g food) and in the control. Cadmium-induced perturbations observed in the gut microbial communities were (1) increased number of morphologically distinct bacterial isolates in the group fed low-cadmium-dosed food (50 microg Cd/g) and reduced number of morphologically distinct bacterial isolates in the group fed high-cadmium-dosed food (250 microg Cd/g) compared with the control, (2) increased or decreased relative frequencies of almost all 25 bacterial units provoked by cadmium-contaminated food, (3) time-dependent increased numbers of gut CFUs in cadmium-fed animals (dose dependence was not observed), and (4) significant changes in community structure described by Shannon equitability indices at lower levels of food contamination (50 microg Cd/g) only. Gut microflora parameters are proposed as additional endpoints in the standardized single-species toxicity test with the terrestrial isopod P. scaber as a means of increasing the ecological relevance of the results. PMID- 11878475 TI - Morphological plasticity in metal-sequestering earthworm chloragocytes: morphometric electron microscopy provides a biomarker. of exposure in field populations. AB - Morphometric measurements of the volume fractions (Vv) of two prominent metal sequestering compartments, chloragosome granules and debris vesicles, were made on transmission electron micrographs of chloragocytes in Dendrodrilus rubidus (Annelida: Oligochaeta: Lumbricidae) from one uncontaminated site and three qualitatively and quantitatively different metalliferous soils associated with disused Pb/Zn mines. We also examined the relative volume fractions of the alimentary wall and attached chloragogenous tissue by light microscopy. Several conclusions relevant to environmental diagnostics were reached: Metal identity and degree of soil pollution exert strong effects on chloragocyte ultrastructure; morphometry by light microscopy indicated that the metal-induced structural effects are characterized by intracellular alterations not accompanied by a significant expansion of the tissue volume; elevated metal burdens can reduce chloragosome Vv and, concomitantly, increase the Vv of their autophagic derivatives, the debris vesicles; the measured cellular alterations were more closely correlated with integrated tissue metal burdens than they were with integrated soil metal burdens; and estimates of tissue metal distribution, derived by combining morphometric data and published whole-worm and chloragosome metal concentrations, suggested that the alimentary wall of earthworms may sequester Pb, Zn, and Cd. Our study shows that cellular changes, directly or indirectly, may yield quantifiable expressions of metal loads bioaccumulated by earthworms. However, the practical use of cellular changes as biomarkers of soil quality probably will require automated light-microscopic morphometry in combination with organelle-specific molecular probes to be cost effective. PMID- 11878476 TI - Influence of metal (Cd and Zn) waterborne exposure on radionuclide (134Cs, 110Ag, and 57Co) bioaccumulation by rainbow trout (Oncorhynchus mykiss): a field and laboratory study. AB - Field and laboratory experiments were carried out to assess the influence of Cd and Zn on the contamination levels of 110Ag, 57Co, and 134Cs in rainbow trout. During a four-week prior exposure phase, two fish groups were held in tanks in the Lot River (France) at a reference (<0.05 microg Cd/L and 68 microg Zn/L) and at a polluted site (1.5 microg Cd/L and 152 microg Zn/L). During a subsequent phase, organisms were brought back to the laboratory, where the radionuclide accumulation and depuration were studied for 14 and 7 d, respectively. During this second phase, the water used in the experiments was brought back from the two sites on the Lot River in order to work under the same chemical conditions. The potential effect of chronic exposure to stable metals on several biomarkers has been explored: Plasma analysis indicated the disruption of certain variables linked to the energetic metabolism and to the maintenance of the ionic balance. In contrast, no significant disruption of the measured enzyme activities was observed. With regard to the bioaccumulation of radionuclides, concentrations in fish exposed to metals are much lower than those in fish from the control group. Various hypotheses are proposed to link fish metabolic profiles due to metal exposure to the radiocontamination of organisms. PMID- 11878477 TI - Relative sensitivity of bull trout (Salvelinus confluentus) and rainbow trout (Oncorhynchus mykiss) to acute copper toxicity. AB - Bull trout (Salvelinus confluentus) were recently listed as threatened in the United States under the federal Endangered Species Act. Past and present habitat for this species includes waterways contaminated with heavy metals released from mining activities. Because the sensitivity of this species to copper was previously unknown, we conducted acute copper toxicity tests with bull and rainbow trout (Oncorhynchus mykiss) in side-by-side comparison tests. Bioassays were conducted using water at two temperatures (8 degrees C and 16 degrees C) and two hardness levels (100 and 220 mg/L as CaCO3). At a water hardness of 100 mg/L, both species were less sensitive to copper when tested at 16 degrees C compared to 8 degrees C. The two species had similar sensitivity to copper in 100-mg/ L hardness water, but bull trout were 2.5 to 4 times less sensitive than rainbow trout in 220-mg/L hardness water. However, when our results were viewed in the context of the broader literature on rainbow trout sensitivity to copper, the sensitivities of the two species appeared similar. This suggests that adoption of toxicity thresholds that are protective of rainbow trout would be protective of bull trout; however, an additional safety factor may be warranted because of the additional level of protection necessary for this federally threatened species. PMID- 11878478 TI - Investigation of effects of trifluoroacetate on vernal pool ecosystems. AB - Trifluoroacetate (CH3COO-, TFA) is a breakdown product of hydrochlorofluorocarbons and hydrofluorocarbons and is released by the heating of Teflon products. Because of its chemical properties, concentrations in evaporative wetlands are predicted to increase with time. This study focused on assessing the impact of this haloacetic acid on vernal pool soil microbial communities as well as vernal pool and wetland plant species. Microbial respiration for three vernal pool soils and an agricultural soil was not affected by TFA exposures (0, 10, 100, 1,000, and 10,000 microg/L), and degradation of TFA by microbial communities was not observed in soils incubated for three months. Trifluoroacetate accumulated in foliar tissue of wetland plant species as a function of root exposure concentration (100 and 1,000 microg/L TFA), and accumulation was found to stabilize or decrease after the second or third month of exposure. Seeds accumulated TFA as a function of root exposure concentration; however, germination success was not affected. No adverse physiological responses, including general plant health and photosynthetic and conductance rates, were observed for root exposures at the TFA concentrations used in this study. Based on the soils and plant species used in this study, predicted TFA concentrations will not adversely affect the development of soil microbial communities and vernal pool plant species. PMID- 11878479 TI - Development of the spray drift task force database for aerial applications. AB - This article is part of a series describing the development of the Spray Drift Task Force (SDTF) database and its application to agricultural chemical exposure risk assessment modeling. The series describes the development of a large generic database (assuming that active ingredient rate is not a factor affecting physical drift) and its use in estimating spray movement immediately following application by aerial methods. The components of the database are described. In agreement with field trials in the open literature, the database shows that the major variables affecting off-target spray deposition are droplet size, spray release position (boom height and length), and wind speed and direction. In addition, secondary parameters that can affect these variables and drift are also discussed. PMID- 11878481 TI - Evaluation of the AgDISP aerial spray algorithms in the AgDRIFT model. AB - A systematic evaluation of the AgDISP algorithms, which simulate off-site drift and deposition of aerially applied pesticides, contained in the AgDRIFT model was performed by comparing model simulations to field-trial data collected by the Spray Drift Task Force. Field-trial data used for model evaluation included 161 separate trials of typical agriculture aerial applications under a wide range of application and meteorological conditions. Input for model simulations included information on the aircraft and spray equipment, spray material, meteorology, and site geometry. The model input datasets were generated independently of the field deposition results, i.e., model inputs were in no way altered or selected to improve the fit of model output to field results. AgDRIFT shows a response similar to that of the field observations for many application variables (e.g., droplet size, application height, wind speed). However, AgDRIFT is sensitive to evaporative effects, and modeled deposition in the far-field responds to wet bulb depression whereas the field observations did not. The model tended to overpredict deposition rates relative to the field data for far-field distances, particularly under evaporative conditions. AgDRIFT was in good agreement with field results for estimating near-field buffer zones needed to manage human, crop, livestock, and ecological exposure. PMID- 11878480 TI - AgDRIFT: a model for estimating near-field spray drift from aerial applications. AB - The aerial spray prediction model AgDRIFT embodies the computational engine found in the near-wake Lagrangian model AGricultural DISPersal (AGDISP) but with several important features added that improve the speed and accuracy of its predictions. This article summarizes those changes, describes the overall analytical approach to the model, and details model implementation, application, limits, and computational utilities. PMID- 11878482 TI - Reduced noise levels in ICU promote rest and healing. PMID- 11878483 TI - Staffing is a concern in telemetry. PMID- 11878484 TI - Duration of action of GpIIb-IIIa antagonists. PMID- 11878485 TI - Prevention of secondary brain injury. PMID- 11878486 TI - Meningococcemia: a critical care emergency. PMID- 11878487 TI - Sedation and pain management in critically ill adults. PMID- 11878488 TI - Managing patients with lumbar drainage devices. AB - All nurses who care for patients with LDDs must have demonstrated initial competency with LDDs and should participate in periodic in-service training to maintain that competency. Clearly an educated critical care nurse is the most essential partner that a patient with an LDD can have for preventing complications and ensuring that the best outcomes for the patient are achieved. Nurses caring for patients with an LDD must have a clear institution-specific policy and procedure available to guide the care of these patients. PMID- 11878489 TI - Overdoses of tricyclic antidepressants: grandchildren and grandparents. AB - TCAs are an extremely toxic source of poisoning in young children. Overdoses of TCAs can cause coma, seizures, hypotension, cardiac arrhythmias, and cardiac arrest. Treatment is directed at rapid assessment, monitoring, support of vital functions, halting drug absorption, and treating CNS and cardiac toxic effects. All children should be monitored for a minimum of 6 hours, and many require admission to a critical care unit. The mainstay of therapy is alkalinization. Intravenous administration of sodium bicarbonate is the preferred treatment for hypotension, shock, and arrhythmias. Blood pH should be monitored and should be maintained between 7.45 and 7.55. More specific drug therapy, cardioversion, or artificial pacing may be required for refractory arrhythmias. Before the child is discharged from the hospital, strategies to reduce the risk of future poisonings should be discussed with the child's family. PMID- 11878490 TI - Vibrio vulnificus sepsis. PMID- 11878491 TI - Messages of professional empowerment from the land of Oz. AB - In the movie adaptation of Baum's book, Dorothy's whole adventure was a dream. But the movie told only a part of the original story. In the print version, Oz was a real place to which Dorothy eventually relocated after 3 more visits; Dorothy moved to Oz permanently in the sixth book (of 13 books) and became a princess. In many ways, critical care professionals are all living in a modern healthcare version of the story of Oz-facing tornadoes, wandering fields of intense expectations, traveling indistinct paths, manipulating wizardly technology, and confronting new enemies. To survive and thrive, we must have brains, heart, and courage. Despite everything that is going on around them, critical care professionals continue to have a sense of mission, a vision of continual improvement and professionalism, and a code of ethics that transcends the complex environment swirling around us. We can begin by creating a personal vision that we wish to achieve. This vision is not an academic exercise but rather a compelling statement that anticipates tomorrow without neglecting today. This vision will require commitment, teamwork, and involvement to make it a reality. Like the end of the original story of Oz, the end of this story is just the beginning. . . . Click your heels together 3 times and say, "It's a new century for critical care nurses!" PMID- 11878492 TI - Sedation and neuromuscular blockade in patients with acute respiratory failure. PMID- 11878493 TI - The Synergy Model in practice. Strengthening nurses' moral agency. PMID- 11878494 TI - The most effective antiseptic method for preparing the skin before insertion of a peripheral intravenous cannula. PMID- 11878495 TI - A peer reviewed system for ICT-based resources: is it possible? PMID- 11878496 TI - The relationship between nurses' personality and stress levels reported when caring for interpersonally difficult patients. AB - The aim of this study was to investigate the self-reported stress in a group of nurses caring for patients with interpersonally difficult behaviours at four Melbourne hospitals. An examination was undertaken of the role played by the personality construct known as Lifestyle, in nurses' cognitive appraisal of interpersonal stressors. One hundred and ten nurses completed the Basic Adlerian Scales for Interpersonal Success, Adult Version (BASIS-A) personality inventory and the Difficult Patient Questionnaire (DPQ) and a subgroup of 40 nurses were subsequently interviewed. The interviews were analysed with the Alexis lexical thematic content analysis computer program using the Harvard III Psychosociological Dictionary. The results revealed no significant correlations between stress and the nurses' demographic or professional backgrounds. However, very specific and significant correlations between certain personality profiles and stress levels were detected. Lexical thematic content analysis indicated that nurses who reported high stress differed significantly from those reporting low stress in the psychological processes they used when caring for interpersonally difficult patients. These findings suggest that the personality construct of Lifestyle plays an important role in regard to the stress response of these nurses. The implications of the findings include the possibility of structuring interventions based on Individual Psychology that may assist nurses to better deal with interpersonal conflict in patient care, and hence to reduce their own stress levels. PMID- 11878497 TI - Violence: part of the job for Australian nurses? AB - According to a recent Australian Institute of Criminology report (1999) the health industry is the most violent industry in Australia. In this paper the authors aim to highlight violence as an important professional issue for Australian nurses that is currently concealed as 'part of the job'. National and international studies bring attention to the severity of the problem for nurses with a particular focus on emergency nurses. Some of the issues identified and discussed include increased waiting times and frustration; increasing use of weapons; inadequate systems of security; culture of silence; inadequate support for emergent mental health needs; lack of reporting; lack of institutional concern and systems of support, and; demands of triage nursing. The nature of workplace violence in emergency departments in New South Wales and South Australia will be explored based on the authors' research. A research pathway to explore national impact and implications of violence for nurses and nursing practice in general will be outlined. PMID- 11878498 TI - Literature reviews: evolution of a research methodology. AB - Literature reviews summarise all past research efforts, and because of this, are increasingly being used as a way of addressing the rapid accumulation of health care information. Because reviews, rather than primary research, are now being used as the basis for many health care decisions, it is important that they are conducted with the same rigour of the primary research. To ensure this rigour, the past two decades have seen a progressive evolution in review methodology, to the point where reviews are now considered research in their own right. Despite this development, the standard of many published reviews remains poor. This paper summarises the development of the literature review methodology and discusses some of the implications for nursing. PMID- 11878499 TI - The use of single-case experimental designs to evaluate nursing interventions for individual clients. AB - Nurses are increasingly called upon to devise interventions for individual clients. These interventions must be effective for the purpose for which they are designed if they are to be considered appropriate for the client for whom they were devised. Single-case research methodology offers the practising nurse an easy to use strategy for assessing the efficacy of individualised interventions, with the controls of an experimental design and the flexibility required for use in applied settings. This paper describes the basic principles underlying the use of these designs and illustrates their application in assessing the efficacy of interventions with individual clients. PMID- 11878500 TI - Danger! nurses at work. PMID- 11878501 TI - Violence in New South Wales emergency departments. AB - In 1999 the International Council of Nurses recognised workplace violence as a significant issue in nursing. During the same year the Australian Institute of Criminology reported that health was the most violent industry. This study examined the nature and extent of violence in NSW hospital emergency departments. Emergency nurses experienced violent incidents in their department, in the wards and outside the hospital setting. Every respondent (n=266) experienced some form of violence at least weekly. Ninety-two incidents involved lethal weapons. Ninety two percent of perpetrators were patients or their relatives, however other staff members were also implicated. Non-reporting of violence is an issue as over 70% of incidents were not referred to authorities. Drugs, alcohol and emergency department waiting times are the most significant predisposing factors. Most emergency nurses are not satisfied with the response of administration to violent incidents within hospitals. PMID- 11878502 TI - A case for chaos theory in nursing. AB - This paper addresses the question of why nurses should understand chaos theory. A critique of the literature is used to demonstrate how chaos theory has been utilised in a number of disciplines, including nursing. Possible applications of chaos theory in nursing are proposed in order to demonstrate where it might assist nurses, in particular researchers, educators and policy makers. The appropriateness of the application of chaos theory as a framework for knowledge generation is also discussed. PMID- 11878503 TI - Nurse practitioners: an evaluation of the extended role of nurses at the Kirketon Road Centre in Sydney, Australia. AB - The aim of the present study was to formally evaluate the effectiveness, professional appropriateness and acceptability of the extended role of the nurse practitioner at the Kirketon Road Centre (KRC) in Sydney, Australia. Data collection consisted of client and staff surveys and case file review by two assessors (one medical and one nursing). This paper will report on one section of this research, namely the case file review section of the study. Total study subjects were 1046 'at risk' youth, sex workers and injecting drug users attending KRC for their primary health care needs between September 1994 and April 1995. Nurse practitioners (NP) saw 613 of the clients who presented over this period. The majority of these clients were women (77.3%). The majority of NP consults were related to STD (51%), gynaecological (17%) and hepatitis (16%) issues. The results demonstrated that nurse practitioners were professionally appropriate in all aspects of expected 'best practice' in over 95% of consultations. PMID- 11878504 TI - A CD-ROM tutorial for physical examination. AB - In 1998 the Committee for University Teaching and Staff Development (CUTSD) awarded a grant of $47,975 to develop an interactive CD-ROM tutorial program to facilitate teaching the process of physical examination of the abdomen, lungs and thorax, to students of nursing. This program was developed to complement current teaching methods and make it possible for tutors to use the available class time to further address students' individual learning needs. It was developed to enable flexible delivery of content, to provide a front seat view of the demonstration of the procedures, and simulated practice opportunities in the safety and privacy of computer simulation. The program was not intended to replace hands-on practice as this learning medium does not address the kinaesthetic component of performing physical examination but it is expected to hasten the development of confidence in practice by strengthening the user's knowledge of the techniques and the sequence of physical examination. Through providing the opportunity to elicit inspection, palpation, percussion and auscultation examination findings in the context of 10 case studies of patients with health problems, it is also expected to facilitate recognition of abnormalities and their significance for health care students who have little clinical experience. The program has been evaluated by senior nursing students for technical problems, effectiveness as a learning aid and user friendliness. Over all, 92% (n=38) of the students considered that the program assisted them to learn physical examination of the abdomen, thorax and lungs and 95% were satisfied with the quality of the product and found that the sounds and images helped their understanding. The content of the program was considered to be logically sequenced, to have assisted understanding, and the case studies were a valuable learning aid. The evaluation data from this trial also indicates that students would like to learn about physical examination of other body systems using this medium. PMID- 11878505 TI - Forensic nursing: a review of the literature. AB - A review of the nursing literature reveals that forensic nursing is an emergent specialty area of practice that has undergone substantive role development in recent years. Forensic nurses have not only begun to write about the challenging and distinctive nature of their practice and their unique practice arrangements, but have commenced a concerted call to action for greater recognition within the nursing profession and correction and criminal justice system. The literature reveals an increasing demand for forensic nursing skills in a range of community and hospital based clinical settings. The problematic nature of caring for forensic clients in both correctional and less restrictive contexts of care remains a salient feature of forensic nurses' accounts of their practice. PMID- 11878507 TI - Leadership for change. PMID- 11878506 TI - Latex sensitivity and allergy: raising awareness in Australia. AB - Natural rubber latex allergy is a public health issue that is yet to be addressed in many Australian hospitals. It is suggested that there is a widespread lack of awareness amongst hospital staff of the implications of this relatively new health problem. Natural rubber latex allergy is a cumulative, serious and incurable occupational health problem and a disabling disease. It can result in chronic illness, disability, loss of career and even death, for nurses and other health professionals. The serious risk that natural rubber latex allergy presents to staff health and patient safety, requires that hospital-wide strategies be developed to address the prevention and management of this condition as a matter of urgency. An overview of the problem of natural rubber latex allergy is presented, and strategies for addressing it suggested. The need for further research into the scope of the problem in the Australian context is emphasised. PMID- 11878508 TI - Enactment of virtue ethics: collaboration between nurse academics and international students in questionnaire design. AB - Increased enrolment of international nursing students in undergraduate and postgraduate programs is predicted as Australian universities compete for students in the globalised university market. This study explored issues impacting on the ability of international nursing students to achieve their study goals at an Australian university (n=29). However, this paper principally describes the design of the instrument by two nursing lecturers and international nursing students (n=10) in a collaborative process, characterised by a pluralistic world view and the enactment of virtue ethics. The myth of the term 'ESL' is critiqued, and challenged. In designing the instrument, we argued it was essential to be cognisant of the conceptual attributes and embedded meanings of language for the unique study population, to ensure face and content validity. PMID- 11878509 TI - Human natural killer cell receptors and signal transduction. AB - Natural killer (NK) cells express numerous receptors, which continually engage with ligands on cell surfaces. Until 1995, only a handful of these receptors were characterized and the molecular basis of NK cell activation was obscure. Recently, considerable advances have been made in characterizing the receptor repertoire on human NK cells. Both activating and inhibitory receptors can transduce positive or negative signals to regulate NK cell cytotoxicity and cytokine release responses. The inhibitory receptors normally predominate in this balance of signals. Certain tumor cells and virally infected cells that lack major histocompatibility complex (MHC) class I molecules, however, can rapidly trigger NK cell activation. The basis of this activation is the loss of negative signals that are normally transmitted by MHC class I-binding inhibitory receptors, and the corresponding domination of activating receptor signals. While ligand specificity for a number of the recently described receptors is still a mystery, their signal transduction properties have begun to be defined. The dynamic crosstalk between these receptors ultimately governs the NK cell activation state. Although the complexities of NK cell signalling are only marginally understood, several overall themes have been defined by characterizing the roles of distinct pathways during NK cell responses. PMID- 11878510 TI - Ig-binding receptors on human NK cells as effector and regulatory surface molecules. AB - The receptors on human natural killer 9NK cells which can specifically bind the Fc portion of immunoglobulin molecules (Fc receptors) have been extensively studied. The best known and studied Fc receptor on human NK cells is FcgammaRIIIa. Interactions of NK cells with IgG antibodies via this receptor are well known to induce a signal transduction cascade and lead to antibody-dependent cell-mediated cytotoxicity (ADCC) as well as release of various cytokines. In addition, interactions with monomeric IgG and FcgammaRIIIa have been demonstrated, which result in negative regulation of NK activity and other immunomodulatory effects. Over the past several years, it has also become increasingly appreciated that human NK cells express a variety of other Fc receptors, including FcmuR, which also can mediate effector and immunoregulatory functions. Also, a novel form of FcgammaR has been demonstrated on human NK cells, termed FcgammaRIIc. Recent molecular studies have shown considerable polymorphism in the genes for FcgammaIIc and the functional consequences are being dissected. This appears to include cross-talk between FcgammaRIIIa and at least some forms of FcgammaRIIc, which may have important functional consequences. PMID- 11878511 TI - Role of TNF family ligands in antitumor activity of natural killer cells. AB - NK cells have the ability to destroy tumor cells by two main cytotoxic pathways, the well-known perforin/granzyme-mediated secretory/necrotic killing and the newly defined TNF family ligand-mediated apoptotic killing. The former mechanism is operative mainly against a few cultured leukemia cell targets, while the latter mediates substantial activity against most tumor cell targets. It also appears from emerging data that the apoptotic mechanism is the main antitumor pathway in vito. This review is focused on the apoptotic mechanism of killing, the molecules and cell signaling pathways involved in this process, and its potential biologic significance along with its relation to the secretory/necrotic cytolytic pathway. PMID- 11878512 TI - Therapeutic activity of NK cells against tumors. AB - While it is generally accepted that natural killer (NK) cells, by killing tumor cells in the circulation, represent a first line of defense against metastases, their therapeutic activity against established tumors has been limited. In this review, we describe studies to improve the therapeutic effectiveness of activated NK cells in both animal models and clinical trials to better understand the biological problems that limit their effectiveness. PMID- 11878513 TI - Ontogeny and expansion of human natural killer cells: clinical implications. AB - Our knowledge of NK cells and their critical role in the innate immune system has increased enormously since their discovery several decades ago. However, it is only within the last 10 years that rational cytokine therapies, such as those utilizing low doses of IL-2, have been successful in expanding NK cells in patients with cancer and/or immunodeficiency. Such experiences in vivo have highlighted the importance of basing immunotherapeutic strategies on the known cellular and molecular properties of the targeted cell population. Recent advances in our understanding of the physiologic factors and events that orchestrate NK cell ontogeny, including IL-15 and receptor tyrosine kinase ligands to c-kit and flt3, provide novel therapeutic possibilities for cytokine therapy. This review summarizes our current understanding of human NK cell ontogeny, and links this knowledge to ongoing and future clinical strategies for the endogenous expansion of NK cells in patients with cancer and/or immunodeficiency. PMID- 11878514 TI - Dexmedetomidine: a real innovation or more of the same? PMID- 11878515 TI - Monitoring medical devices: the need for new evaluation methodology. PMID- 11878516 TI - Pain relief after thoracotomy. PMID- 11878517 TI - Comparison between dexmedetomidine and propofol for sedation in the intensive care unit: patient and clinician perceptions. AB - The alpha2 agonist dexmedetomidine is a new sedative and analgesic agent which is licensed in the USA for post-operative intensive care sedation. We compared dexmedetomidine with propofol in patients requiring sedation in intensive care. Twenty adult patients expected to require a minimum of 8 h artificial ventilation after surgery were randomized to receive sedation with either dexmedetomidine or propofol infusions. Additional analgesia, if required, was provided by an alfentanil infusion. Depth of sedation was monitored using both the Ramsay sedation score (RSS) and the bispectral index (BIS). Cardiovascular, respiratory, biochemical and haematological data were obtained. Patients' perceptions of their intensive care stay were assessed using the Hewitt questionnaire. Sedation was equivalent in the two groups [median (interquartile range): RSS, propofol group 5 (4-5), dexmedetomidine group 5 (4-6) (P=0.68); BIS, propofol group 53 (41-64), dexmedetomidine group 46 (36-58); P=0.32], but the propofol group received three times more alfentanil compared with patients sedated with dexmedetomidine [2.5 (2.2-2.9) mg h(-1) versus 0.8 (0.65-1.2) mg h(-1) (P=0.004)]. No differences were found in arterial pressures between the groups, but heart rate was significantly lower in the dexmedetomidine group [mean (SD) 75 (6) vs 90 (4) beats min(-1)]. Extubation times were similar and rapid with the use of both sedative agents [median (range) 28 (20-50) and 29 (15-50) min (P=0.63) respectively for the propofol and dexmedetomidine groups]. No adverse events related to the sedative infusions occurred in either group. Despite ventilation and intubation, patients sedated with dexmedetomidine could be easily roused to cooperate with procedures (e.g. physiotherapy, radiology) without showing irritation. From the clinician's and patient's perspectives, dexmedetomidine is a safe and acceptable sedative agent for those requiring intensive care. The rate pressure product is reduced in patients receiving dexmedetomidine, which may protect against myocardial ischaemia. Dexmedetomidine reduces the requirement for opioid analgesia. PMID- 11878518 TI - Influence of equianaesthetic concentrations of nitrous oxide and isoflurane on regional cerebral blood flow, regional cerebral blood volume, and regional mean transit time in human volunteers. AB - Nitrous oxide and isoflurane have cerebral vasodilatory effects. The use of isoflurane in neuroanaesthesia is widely accepted, whereas the use of nitrous oxide in neuroanaesthesia is still the subject of debate. In the present study, contrast-enhanced magnetic resonance (MR) perfusion measurement was used to compare the effects of 0.4 MAC nitrous oxide (n=9) and 0.4 MAC isoflurane (n=9) on regional cerebral blood flow (rCBF), regional cerebral blood volume (rCBV) and regional mean transit time (rMTT) in spontaneously breathing human volunteers. Nitrous oxide increased rCBF and rCBV in supratentorial regions more than did isoflurane. Isoflurane, by contrast, increased rCBF and rCBV in basal ganglia more than did nitrous oxide. An increased rMTT was caused by a relatively greater increase in rCBV than in rCBF supratentorially by isoflurane and infratentorially by nitrous oxide. In conclusion, nitrous oxide increases rCBF and rCBV predominantly in supratentorial grey matter, whereas isoflurane increases rCBF and rCBV predominantly in infratentorial grey matter. PMID- 11878519 TI - Cerebral effects and blood sparing efficiency of sodium nitroprusside-induced hypotension alone and in combination with acute normovolaemic haemodilution. AB - The combined reduction of oxygen-carrying capacity and perfusion pressure during the combination of acute normovolaemic haemodilution (ANH) and controlled hypotension (CH) raises concerns of hypoperfusion and ischaemic injury to the brain. Forty-two patients undergoing radical prostatectomy were prospectively allocated to receive CH induced by sodium nitroprusside (mean arterial pressure (MAP) 50 mm Hg), a combination of CH+ANH (post-ANH haematocrit 29%; intraoperative MAP 50 mm Hg), or standard anaesthesia (control). Serum levels of the brain-originated proteins neuron-specific enolase (NSE) and protein S-100, blood loss, transfusion requirements, adverse effects, and postoperative recovery profile were compared among the three groups. Intraoperative blood loss in the CH group (mean (SD)) (788 (193) ml) and CH+ANH group (861 (184) ml) was significantly less than in the control group (1335 (460) ml). Significantly fewer total units of allogeneic packed red blood cells (PRBC) were transfused in the patients receiving hypotensive anaesthesia (CH, 3 units; CH+ANH, 2 units; control, 17 units). There was no difference in immediate postoperative recovery profile among the three groups as determined by the emergence from anaesthesia and time to discharge from the postanaesthesia care unit. Serum S-100 protein concentrations increased significantly in all groups from baseline to peak concentrations 2 h postoperatively (CH 0.25 (0.11) microg litre(-1); CH+ANH 0.31 (0.12) microg litre(-1); control 0.31 (0.10) microg litre(-1)). A return to baseline values was seen within 24 h postoperatively in all patients. No changes in NSE concentrations were seen. Our observations suggest that CH and CH+ANH were effective in reducing blood loss and transfusion requirements in patients undergoing radical prostatectomy. Increased serum S-100 protein concentrations imply a disturbance in astroglial cell membrane integrity and an increased endothelial permeability of the blood-brain barrier. There were no associations between serum S-100 protein or NSE and adverse cognitive effects. Further work needs to be done to determine the prognostic importance of S-100 protein and NSE as surrogate variables of postoperative cerebral complications. PMID- 11878520 TI - Effects of minor surgery and endotracheal intubation on postoperative breathing patterns in patients anaesthetized with isoflurane or sevoflurane. AB - We studied the effects of minor surgery and endotracheal intubation on postoperative breathing patterns. We measured breathing patterns and laryngeal resistance during the periods immediately before intubation (preoperative) and immediately after extubation following minor surgery (postoperative) in eight patients anaesthetized with sevoflurane and eight patients anaesthetized with isoflurane, breathing spontaneously through a laryngeal mask airway at a constant end-tidal anaesthetic concentration (1.0 MAC). In both sevoflurane-anaesthetized and isoflurane-anaesthetized patients, expiratory time was reduced and inspiratory and expiratory laryngeal resistance increased after surgery. In sevoflurane-anaesthetized patients, occlusion pressure (P0.1) increased without changes in inspiratory time (T(I)). Occlusion pressure did not change and T(I) was greater in isoflurane-anaesthetized patients after surgery. Minor surgery may have a small but significant influence on breathing and increased laryngeal resistance following endotracheal intubation may modulate these changes. The difference in breathing pattern between sevoflurane and isoflurane may be a result of different responses of the central nervous system to different anaesthetics in the presence of increased laryngeal resistance. PMID- 11878521 TI - Use of multi-plane transoesophageal echocardiography in visualization of the main hepatic veins and acquisition of Doppler sonography curves. Comparison with the transabdominal approach. AB - The role of multi-plane transoesophageal echocardiography (TOE) in the visualization of the three main hepatic veins and acquisition of Doppler sonography curves has not been established. We have studied this diagnostic option of TOE in 34 patients during general anaesthesia. The findings were compared with the results of conventional transabdominal sonography (TAS). Using TOE, each of the three main hepatic veins could be visualized in all patients. In contrast, TAS allowed adequate two-dimensional visualization of the right, middle, and left hepatic vein in only 97%, 85%, and 61% of the patients, respectively. Adequate Doppler tracings of the right and middle hepatic vein could be obtained in 100% and 97% of the patients by TOE and in 91% and 50% of the patients by TAS. Doppler tracings of the left hepatic vein could only be acquired in 18% of the patients by TOE, but in 47% of the patients by TAS. As blood flow may be calculated from the diameter of the vessel, velocity time integral of the Doppler curve and heart rate, TOE may provide an interesting non invasive tool to monitor blood flow in the right and middle hepatic vein. PMID- 11878522 TI - Comparative efficacy and safety of remifentanil and fentanyl in 'fast track' coronary artery bypass graft surgery: a randomized, double-blind study. AB - This multi-centre, parallel group, randomized, double-blind study compared the efficacy and safety of high-dose remifentanil administered by continuous infusion with an intermittent bolus fentanyl regimen, when given in combination with propofol for general anaesthesia in 321 patients undergoing elective coronary artery bypass graft surgery. A significantly lower proportion of the patients who received remifentanil had responses to maximal sternal spread (the primary efficacy endpoint) compared with those who received fentanyl (11% vs 52%; P<0.001). More patients who received remifentanil responded to tracheal intubation compared with those who received fentanyl (24% vs 9%; P<0.001). However, fewer patients who received remifentanil responded to sternal skin incision (11% vs 36%; P<0.001) and sternotomy (14% vs 60%; P <0.001). Median time to extubation was longer in the subjects who received remifentanil than for those who received fentanyl (5.1 vs 4.2 h; P=0.006). There were no statistically significant differences between the two groups in the times for transfer from intensive care unit or hospital discharge but time to extubation was significantly longer in the remifentanil group. Overall, the incidence of adverse events was similar but greater in the remifentanil group with respect to shivering (P<0.049) and hypertension (P<0.001). Significantly more drug-related adverse events were reported in the remifentanil group (P=0.016). There were no drug-related adverse cardiac outcomes and no deaths from cardiac causes before hospital discharge in either treatment group. PMID- 11878523 TI - Effect of epidural bupivacaine vs combined epidural bupivacaine and morphine on gastrointestinal function and pain after major gynaecological surgery. AB - In a double-blind study, we investigated the effects of postoperative epidural local anaesthetic, with or without addition of epidural morphine, on postoperative pain and gastrointestinal function in patients scheduled for radical hysterectomy and pelvic lymphadenectomy. Forty patients were randomized into two study groups: 48-h postoperative epidural 0.2% bupivacaine 8 ml h(-1) (bupi group) or 48-h postoperative epidural 0.2% bupivacaine/morphine 50 microg at 4 ml h(-1) (bupi/morph group). Patients were observed for at least 96 h after surgery. No differences in pain at rest, during cough or mobilization were observed. Patients in the bupi group requested a significant greater amount of supplementary analgesics, but times to first flatus and defaecation were reduced compared with patients in the bupi/morph group. Itching was a significant problem in patients in the bupi/morph group. No differences in postoperative nausea and vomiting, mobilization or time to discharge from hospital were observed between groups. The addition of morphine to postoperative epidural bupivacaine has only limited effect on pain relief and increases time to normalization of gastrointestinal function. PMID- 11878524 TI - Intrathecal ropivacaine or bupivacaine with fentanyl for labour. AB - Combined spinal-epidural (CSE) is widely used to provide pain relief in labour while minimizing motor blockade. Aiming to further reduce associated motor weakness, we compared ropivacaine 2.5 mg in the intrathecal injection with a standard bupivacaine CSE in a double-blind study. Forty women were randomized to receive either bupivacaine 2.5 mg or ropivacaine 2.5 mg intrathecally, both with fentanyl 0.025 mg. There were no significant differences between the groups regarding the onset, duration or quality of analgesia or the level of sensory block attained. Forty per cent of the women (8/20) receiving bupivacaine developed detectable motor block compared with only 5% (1/20) in the ropivacaine group (P<0.05). Vibration sense was impaired in one woman in each group. Adverse effects did not differ between groups. We conclude that intrathecal ropivacaine 2.5 mg in combination with fentanyl 0.025 mg as part of a CSE technique provides rapid and safe analgesia for labour as effective as that achieved with bupivacaine 2.5 mg and with significantly less motor block. PMID- 11878525 TI - Spinal anaesthesia with 0.5% hyperbaric bupivacaine in elderly patients: effects of duration spent in the sitting position. AB - Sixty patients, aged 65-84 yr, undergoing minor urological surgery under spinal anaesthesia remained sitting for 2 (group 1, n = 15), 5 (group 2, n = 15), 10 (group 3, n = 15), or 20 (group 4, n = 15) min after completion of the subarachnoid administration of 3 ml of a 0.5% hyperbaric bupivacaine solution. They were then placed in the supine position. Analgesia levels were assessed bilaterally using pinprick. Motor block was scored using a 12-point scale. Systolic and diastolic arterial pressures and heart rate were also recorded. Twenty minutes after the injection the upper analgesia levels were lower (P<0.05) in group 4 (median T9.0) than in the groups 1-3 (medians T6.6-T8.5). The highest obtained levels (medians T5.7-T8.0) did not differ between the groups, but occurred later (P<0.05) in group 4 (median 35 min) than in groups 1-3 (medians 19 24 min). There were no significant differences in the maximum degree of motor block or haemodynamic changes between the four study groups. PMID- 11878526 TI - Intrathecal ropivacaine for total hip arthroplasty: double-blind comparative study with isobaric 7.5 mg ml(-1) and 10 mg ml(-1) solutions. AB - This study was designed to evaluate the efficacy and safety of two concentrations of intrathecal ropivacaine, 7.5 and 10 mg ml(-1), in patients undergoing total hip arthroplasty. One hundred and four patients, ASA I-III, were randomized to receive an intrathecal injection of one of two concentrations of isobaric ropivacaine. Group 1 (n=51) received 2.5 ml of 7.5 mg ml(-1) ropivacaine (18.75 mg). Group 2 (n=53) received 2.5 ml of 10 mg ml(-1) ropivacaine (25 mg). The onset and offset of sensory block at dermatome level T10, maximum upper and lower spread of sensory block and the onset, intensity and duration of motor block were recorded, as were safety data. Onset of motor and sensory block was rapid with no significant differences between the two groups. The median time of onset of sensory block at the T10 dermatome was 2 min (range 1-25 min) in Group 1 and 2 min (range 1-21 min) in Group 2. The median duration of sensory block at the T10 dermatome was 3.0 h (range 0.5-4.2 h) in Group 1 and 3.4 h (1.1-5.9 h) in Group 2 (P=0.002). The median duration of complete motor block was significantly prolonged (P<0.05) in Group 2 compared with Group 1 (1.9 vs 1.2 h, respectively). Anaesthetic conditions were excellent in all but one patient. Intrathecal ropivacaine, in doses of 18.75 and 25 mg, was well tolerated and provided effective anaesthesia for total hip arthroplasty. PMID- 11878527 TI - Inhalation anaesthetics increase heart rate by decreasing cardiac vagal activity in dogs. AB - Inhalation anaesthetics decrease heart rate in isolated hearts but mostly increase heart rate in the intact organism, although most inhibit sympathetic drive. Differences in the degree of increase in heart rate between agents may be related to differences in their vagolytic action. To test this hypothesis, we studied the effects of halothane (H), isoflurane (I), enflurane (E), sevoflurane (S) and desflurane (D) [1-3 MAC (minimum alveolar concentration)] on heart rate and heart rate variability (HRV) as a measure of cardiac vagal activity in seven dogs. HRV was analysed in the time domain as the standard deviation of the RR interval (SDNN) and in the frequency domain as power in the high-frequency (HF, 0.15-0.5 Hz) and low-frequency (LF, 0.04-0.15 Hz) ranges. Heart rate increased with anaesthetic concentration and there were corresponding decreases in SDNN, HF power and LF power. Heart rate increased most with D (+40 beats min(-1)), least with H (+8 beats min(-1)) and to an intermediate extent with S, I and E. SDNN and HF power, as measures of vagal activity, changed in the opposite direction and decreased in the same order as heart rate increased. However, SDNN and HF power correlated significantly with heart rate [r=-0.81 (0.04) and -0.81 (0.03) respectively] and were independent of the anaesthetic and its concentration (P<0.05). Consistent with our hypothesis, these results suggest that differences between agents in the degree of increase in heart rate are explained by differences in their vagolytic action. PMID- 11878528 TI - Structure, conformation, and action of neuromuscular blocking drugs. PMID- 11878529 TI - ADCON-L and hypotension during lumbar microdiscectomy. AB - ADCON-L anti-adhesion gel is used as an antifibrotic agent in surgery for herniated lumbar disc. We report seven patients undergoing lumbar microdiscectomy, in whom ADCON-L gel was applied to the nerve root before closure of the surgical site. After the administration of ADCON-L, tachycardia and hypotension were noted, and were quickly and effectively reversed by i.v. fluids and ephedrine. Since its routine introduction into our practice, we have given ADCON-L on 212 occasions, with an incidence of adverse haemodynamic reactions of 3.3%. Adverse cardiovascular reactions do not appear to have been reported before. PMID- 11878530 TI - Randomized controlled study of colloid preload before spinal anaesthesia for caesarean section. AB - We randomized women having elective Caesarean section to receive either no preload (control group, n=33) or 4% gelatin solution (Gelofusine) 15 ml kg(-1) (colloid group, n=35) i.v. before spinal anaesthesia. Intravenous metaraminol was titrated at 0.25-0.75 mg min(-1) to maintain systolic arterial pressure (SAP) in the target range 90-100% of baseline after the spinal injection. The control group required more vasopressor in the first 10 min [median 1.7 (range 0-2.9) mg vs 1.4 (0-2.8), P=0.02] at a greater maximum infusion rate [0.5 (0-0.75) vs 0.25 (0-0.5) mg min(-1), P=0.0005] and had a lower minimum SAP [90 (51-109) vs 101 (75 127) mm Hg, P=0.006] than the colloid group. Nausea was less frequent in the colloid group (6 vs 24%) but neonatal outcome was similar in the two groups. Colloid preload improved haemodynamic stability but did not affect neonatal outcome when arterial pressure was maintained with an infusion of metaraminol during spinal anaesthesia for Caesarean section. PMID- 11878531 TI - Subclinical hepatic dysfunction in laparoscopic cholecystectomy and laparoscopic colectomy. AB - Laparoscopic surgery causes a reduction in hepatic blood flow due to a number of factors, including raised intra-abdominal pressure, the neurohumoral response to surgical stress and the effect of patient position. The clinical significance of the phenomenon is not fully understood. Plasma concentrations of alcohol dehydrogenase (AD) and glutathione S-transferase (GST), which are concentrated in the centrilobular acinus of the liver, sensitively reflect hepatic hypoperfusion, and can be used to monitor reductions in hepatic blood flow. We compared perioperative AD, GST, aspartate aminotransferase (AST, normal range 14-32 IU litre(-1)) and alanine aminotransferase (ALT, normal range 8-41 U litre(-1)) concentrations in patients undergoing laparoscopic cholecystectomy or laparoscopic colectomy to study how patient position and surgical manipulation of the liver affect hepatocellular integrity during laparoscopy. There were significant postoperative increases in AD and GST in the cholecystectomy group [mean (SD) peak concentration 10.8 (4.7) U litre(-1) and 113 (55) microg litre( 1) respectively]. Although the duration of pneumoperitoneum was longer in the colectomy group, there were no comparable perioperative increases in AD and GST in this group [peak concentration 4.0 (4.0) U litre(-1) and 33 (35) microg litre( 1) respectively]. AST and ALT on the first postoperative day were significantly higher in the laparoscopic cholecystectomy group (41 and 34 U litre(-1) respectively) than in the laparoscopic colectomy group (24 and 18 U litre(-1); P<0.05 for each). These results indicate that patient position and the effects of surgical manipulation of the liver affect perioperative hepatic perfusion significantly. PMID- 11878532 TI - Effects of the auditory stimuli of an auditory evoked potential system on levels of consciousness, and on the bispectral index. AB - Investigators in the field of depth of anaesthesia monitoring sometimes measure the auditory evoked potential (AEP) and the Bispectral Index (BIS) concurrently. However, the auditory stimuli required to generate an AEP may increase the level of consciousness, and cause an increase in the BIS. They may also alter the BIS by producing phase-locked harmonics in the surface electroencephalogram. The aim of this study was to determine if AEP stimuli have clinically significant effects on levels of consciousness and BIS values during sedation and general anaesthesia. Ten healthy adult patients were studied by measuring and recording the BIS for 6 epochs of 5 min each. The first 3 epochs took place during steady state sedation, during which time the Observer's Assessment of Awareness/Sedation (OAA/S) score was also measured. The second 3 epochs took place during steady state anaesthesia. During alternate epochs, patients were subjected to the auditory stimuli generated by an AEP system. The auditory stimuli were not associated with a change in BIS values (during sedation and anaesthesia) or OAA/S scores (sedation). PMID- 11878533 TI - Long-term consequences of repeated pentobarbital anaesthesia on choice reaction time performance in ageing rats. AB - Recent studies have suggested that anaesthesia may be a factor in cognitive decline with age. We examined the effect of repeated (eight times) anaesthesia with pentobarbital on reaction time performance in rats in a longitudinal study. Treated rats had faster response times and made more premature responses than the control rats when they were older than 21 months. The results suggest that repeated anaesthesia during the lifespan can lead to an increase in impulsivity, as assessed by a choice reaction time test, during the later stages of life in the rat. These findings support the theory that repeated anaesthesia is a biological factor that affects cognitive ageing. PMID- 11878534 TI - Grand mal convulsion and plasma concentrations after intravascular injection of ropivacaine for axillary brachial plexus blockade. AB - We report a patient to whom ropivacaine 1.1 mg kg(-1) was administered for brachial plexus blockade and who developed grand mal convulsions because of inadvertent i.v. injection. No symptoms of cardiovascular toxicity occurred. Venous blood samples were taken 15, 45, 75 and 155 min after the injection. The measured total plasma concentrations of ropivacaine were 3.3, 1.6, 1.2 and 1.0 mg litre(-1) respectively. Initial plasma concentration after the end of the injection period was estimated at 5.75 mg litre(-1) using a two-compartment pharmacokinetic model. PMID- 11878535 TI - The Montgomery T-tube: anaesthetic problems and solutions. AB - The Montgomery T-tube is a device used as a combined tracheal stent and an airway after laryngotracheal surgery. The device is used mostly in specialist centres for head and neck surgery, and therefore, many anaesthetists may be unfamiliar with its use. The Montgomery T-tube presents the anaesthetist with challenges both during its surgical insertion when acute loss of the airway might occur and also during induction of anaesthesia in patients who have such a tube in situ. Anaesthetists who are unfamiliar with the tube may have to resort to ingenious ways of coping with the problems of a shared airway with a T-tube, which does not have a suitable adaptor for a standard catheter mount, as well as controlling and maintaining ventilation through the device. Safe management of such patients requires careful planning. We describe the anaesthetic management of two cases to illustrate the problems associated with Montgomery tubes. PMID- 11878536 TI - Unnecessary emergency caesarean section due to silent CTG during anaesthesia? AB - We present a case of a probably unnecessary Caesarean section due to misinterpretation of the cardiotocography (CTG) trace during general anaesthesia. A 27-yr-old patient in her 30th week of an uneventful, normal first pregnancy presented with a deep venous thrombosis in the pelvic region. She was to undergo an emergency thrombectomy under general anaesthesia. During the operation, the CTG showed a lack of beat-to-beat heart rate variation (silent pattern CTG) with normal fetal heart rate. This silent CTG pattern was probably a result of the effect of general anaesthesia on the fetus. The CTG pattern was interpreted as indicating fetal distress, and an emergency Caesarean section was performed after the thrombectomy. The infant was apnoeic and had to be resuscitated and admitted to the neonatal intensive care unit. The pH at delivery was 7.23 and the baby was extubated 2 days later. Mother and child recovered without short-term sequelae. In the absence of alternative explanations, reduced fetal beat-to-beat variability with a normal baseline heart rate during general anaesthesia is probably normal. PMID- 11878537 TI - Pathophysiological role of the serotonin system in malignant hyperthermia. PMID- 11878538 TI - How can we improve the outcome of oesophagectomy? PMID- 11878539 TI - Perioperative risk factors for acute lung injury following elective oesophagectomy. PMID- 11878540 TI - Diagnostic confusion caused by dapsone-induced methaemoglobinaemia. PMID- 11878541 TI - Cancellation of hypertensive patients. PMID- 11878542 TI - Is it safe to artificially ventilate a paralysed patient through a laryngeal mask? PMID- 11878543 TI - An unusual case of airway obstruction. PMID- 11878544 TI - Rocuronium: high risk for anaphylaxis? PMID- 11878545 TI - Perioperative bradycardia. PMID- 11878546 TI - Using action research within a school of nursing: exposing tensions in ideologies. AB - This paper examines and critically reflects on a recent curriculum evaluation that took place in 1999 within a school of nursing. Critical theory, and in particular action research, was chosen as an approach for the research. The method aimed to foster participation and reveal and problematise aspects of nursing education which had become taken for granted. Through the process of action research a number of tensions and challenges were revealed. The exposed tensions and challenges are discussed and reframed so that they offer potential for renewed commitment to nursing education, rather than continued constraint and conformity. PMID- 11878547 TI - New mother groups as a social network intervention: consumer and maternal and child health nurse perspectives. AB - Maternal and child health nurses in two outer urban local government areas in Melbourne, Australia were interviewed about how they facilitated first-time parent groups. Groups were offered to all first time mothers and almost two thirds of mothers joined a group. The groups ran for approximately eight sessions and provided infant-focussed parent education and social contact. Women who joined the groups were followed up 18 months to two years later to determine the degree to which these groups continued to meet on their own accord and the extent to which they had become self-sustaining social networks. The study found a very high level of continuation, suggesting that providing such programs may be an important vehicle for enhancing social support during the transition to parenthood and thus be a useful primary prevention strategy. PMID- 11878548 TI - Engendering change: empowering nurse academics to take part in a university research culture. AB - Any school of nursing, which is building upon a college-based teaching culture to create and maintain a viable research culture within a university, must build from within its own resources. This paper outlines a strategic approach to create a research culture in one such school. We describe the empowerment philosophy based on critical and feminist approaches that underpinned our strategy in transforming what was a teaching based college of advanced education culture to that of a university in which both research and teaching are required of its staff. A climate to facilitate change was created and a research support structure was put in place. The success of the strategies can be assessed by the increased participation in research activities, enhanced productivity and evidence of increasing confidence of staff. PMID- 11878549 TI - Educating nurses to protect the past or to advance health care?: A polemic. AB - Internationally, nursing is challenged by recruitment and retention issues. On the one hand nursing is expanding and extending practice whilst on the other, lesser skilled workers are undertaking many roles which were previously fulfilled by nurses. This opinion paper contextualises the current debate and outlines many of the reasons that have been proffered for the apparent shortage. It then progresses to suggest strategies that may ensure nursing remains relevant to contemporary health care. Essentially a presentation of the authors' views, the intent of the paper is to generate debate. PMID- 11878550 TI - Disease management: coming to a hospital near you. PMID- 11878551 TI - The development, validity and reliability of the Hospital in the Home Dependency Scale (HDS). AB - The aim of this study was to develop and investigate the validity and reliability of the Hospital-in-the-Home (HITH) Dependency Scale (HDS). The HDS is a new instrument designed to measure the dependency of HITH patients. It calculates an overall dependency level by rating four dimensions of the provision of HITH nursing care. Specifically, these dimensions are the complexity of assessment, complexity of treatment, time taken to provide the treatment, and the frequency of treatment. The results of testing the HDS suggest that it is valid in measuring adult medical and surgical HITH patient dependency. The scale demonstrated strong stability over time in test retest procedures over a one month period (r = 0.80, p <0.01) and internal consistency (Cronbach's alpha = 0.72). We conclude that the HDS is a valid, reliable instrument that is quick and easy to use in the HITH setting. PMID- 11878553 TI - Planned adolescent pregnancy: what they needed. AB - Adolescent pregnancy represents a significant portion of all pregnancies that occur in the United States. However, little is known about adolescents who plan to become pregnant. This phenomenologic investigation attempts to fill the gap in the current literature by addressing this question. Eight adolescent girls between the ages of 14 and 17 were interviewed regarding their experiences planning pregnancy. Participants were recruited from a prenatal clinic in a large tertiary care center in New York City. Themes that emerged from the interviews were related to planning the pregnancy and to the pregnancy. Themes related to planning the pregnancy were further classified as either needs or wants of the adolescent participants. The themes related to adolescent needs are presented in this article and include: that certain important criteria were met for pregnancy including financial, relationship, and age-related goals; planning involved the boyfriend at least to some extent; environmental issues; and the need for stability. Each theme is discussed in depth and recommendations for practice are addressed. PMID- 11878552 TI - Sexual health practices of homeless youth: a model for intervention. AB - This article presents a conceptual model of sexual health practices and outcomes among homeless adolescents. The model provides a framework for understanding what is known about homeless youth and how community-based interventions might be developed and tested to curb the rising number of sexually transmitted diseases among this vulnerable population. Research measures are presented and discussed to promote further development and testing of the model for interventions. PMID- 11878554 TI - Child health education for the foreign-born parent. AB - Providing child health education for the foreign-born parent presents unique concerns related to language and culture. An innovative approach in a transcultural format used a presentation of basic child health information in English, with translators as facilitators. Foreign-born parents who need partial or complete language interpretation have ready access to translation support. The parents offer questions, comments, suggestions and evaluate the presentation through the translator. Each presentation can accommodate more than one language, since participants are grouped with the appropriate translator. The presentation is done in English and paced to allow for translation to be completed as material is offered. This type of presentation allows discussion of child health in a forum apart from the pediatric care setting. Because the presentation is the only focus, parents do not have the additional concern of immediate care of the child added to communication issues. Vocabulary relative to health care is developed from English into the parents' primary language, with the support of the translator. The pediatric nurse presenter has an opportunity to review health care practices that parents prefer, as well as interpret safety and efficacy. PMID- 11878556 TI - Archaeal DNA replication: spotlight on a rapidly moving field. AB - The replication of DNA is a fundamental step in the cell cycle, which must be coordinated with cell division to ensure that the daughter cells inherit the same genomic material as the parental cell. The recently published complete genome sequences of some archaeal species together with preliminary biochemical studies suggest that the Archaea quite likely duplicate their chromosome by using replication machinery that seems to be a simplified version of the eukaryotic machinery, although their metabolic facets and their cellular morphology are prokaryotic-like. This review is focused on recent progress on the structural and functional analysis of proteins and enzymes involved in the initiation and elongation steps of DNA replication in Archaea. Differences between the genome replication apparatus of the Euryarchaea and the Crenarchaea (the two main phylogenetic divisions of the Archaea domain) are highlighted. PMID- 11878555 TI - Self-medication with common household medicines by young adolescents. AB - Our study examines the self-medication practices of children ages 10-14. We developed a questionnaire that addressed several aspects of such behavior. A convenience sample of subjects (n = 86) completed the questionnaires in school settings in a northeastern metropolitan area. The data showed that a vast majority of the adolescents (89%) have access to the medicine in their households: 36% of them medicated themselves the last time they had any medicine. Most of their medication choices were appropriate, except in the area of gastrointestinal problems. There is a need for further investigation of this issue and incorporating the findings into the practice of pediatric nurses. PMID- 11878557 TI - Investigation into a stress-inducible promoter region from Marinococcus halophilus using green fluorescent protein. AB - A 480-bp Marinococcus halophilus DNA-fragment upstream of the ectoine genes ectABC was linked to the reporter gene gfp(uv) in the stress probe plasmid pBRGFP(uv). On the basis of green fluorescence emitted after application of different types of stress to recombinant Escherichia coli, the DNA sequence was characterized as a stress-responding promoter region. In E. coli, osmotic stress resulted in an increasing fluorescence intensity, whereas the effects of heat or cold were insignificant, suggesting that transcription and/or translation is controlled mainly by an osmotic stress/low-water stress response. Additionally, the stress-counteracting effects of the compatible solutes ectoine and betaine were demonstrated. PMID- 11878558 TI - The thermostability of DNA-binding protein HU from mesophilic, thermophilic, and extreme thermophilic bacteria. AB - Based on primary structure comparison between four highly homologous DNA-binding proteins (HUs) displaying differential thermostability, we have employed in vitro site-directed mutagenesis to decipher their thermostability mechanism at the molecular level. The contribution of the 11 amino acids that differ between the thermophilic HUBst from Bacillus stearothermophilus (Tm = 61.6 degrees C) and the mesophilic HUBsu from Bacillus subtilis (Tm = 39.7 degrees C) was evaluated by replacing these amino acids in HUBst with their mesophilic counterparts. Among 11 amino acids, three residues, Gly-15, Glu-34, and Val-42, which are highly conserved in the thermophilic HUs, have been found to be responsible for the thermostability of HUBst. These amino acids in combination (HUBst-G15E/E34D/V42I) reduce the thermostability of the protein (Tm = 45.1 degrees C) at the level of its mesophilic homologue HUBsu. By replacing these amino acids in HUBsu with their thermophilic counterparts, the HUBsu-E15G/D34E/142V mutant was generated with thermostability (Tm = 57.8 degrees C) at the level of thermophilic HUBst. Employing the same strategy, we generated several mutants in the extremely thermophilic HUTmar from Thermotoga maritima (Tm = 80.5 degrees C), and obtained data consistent with the previous results. The triplet mutant HUTmar G15E/E34D/V421 (Tm = 35.9 degrees C) converted the extremely thermophilic protein HUTmar to mesophilic. The various forms of HU proteins were overproduced in Escherichia coli, highly purified, and the thermostability of the mutants confirmed by circular dichroism spectroscopy. The results presented here were elucidated on the basis of the X-ray structure of HUBst and HUTmar (our unpublished results), and their mechanism was proposed at the molecular level. The results clearly show that three individual local interactions located at the helix-turn-helix part of the protein are responsible for the stability of HU proteins by acting cooperatively in a common mechanism for thermostability. PMID- 11878559 TI - Marinospirillum alkaliphilum sp. nov., a new alkaliphilic helical bacterium from Haoji soda lake in Inner Mongolia Autonomous Region of China. AB - A new helical, alkaliphilic, gram-negative, chemoorganotrophic bacterium designated strain Z4T was isolated from Haoji soda lake in Inner Mongolia Autonomous Region, China. The isolate grows at salinities between 0.2% and 5.0% (w/v) NaCl and pH range 7.0-11.0, with an optimum at 2.0% (w/v) NaCl and pH 9.5. Its growth temperature ranges from 8 degrees to 49 degrees C with an optimum at 37 degrees C. The G+C content of the DNA is 46.8 mol%. The major isoprenoid quinone is ubiquinone 8 (Q-8). Phylogenetic analyses based on 16S rDNA sequence comparison indicates that strain Z4T is a member of the genus Marinospirillum. Phenotypic features and DNA-DNA homology of less than 20% with the described species of Marinospirillum support the view that strain Z4T represents a new species of the genus Marinospirillum. Strain Z4T (= AS 1.2746) is proposed as the type strain of a new species, named Marinospirillum alkaliphilum sp. nov. PMID- 11878560 TI - Sulfolobus tokodaii sp. nov. (f. Sulfolobus sp. strain 7), a new member of the genus Sulfolobus isolated from Beppu Hot Springs, Japan. AB - The taxonomic position of a thermoacidophilic crenarchaeote Sulfolobus sp. strain 7, previously isolated from the Beppu Hot Springs in the geothermal area of Kyushu Island, Japan, was investigated by cloning and sequencing, by phylogenetic analysis of the 16S rRNA gene sequence, by DNA-DNA homology with similar species, and by biochemical characterization of the isolate. This isolate is an obligate aerobe and grows optimally at 80 degrees C and pH2.5-3 under aerobic and chemoheterotrophic growth conditions by aerobic respiration rather than simple fermentation. In conjunction with the phenotypic properties, the present phylogenetic analysis based on the 16S rRNA gene sequence and DNA-DNA hybridization experiments indicate that this isolate is related to the described Sulfolobus taxon and should be considered a novel species of the genus. We propose that this isolate is a novel species of the genus Sulfolobus that we name Sulfolobus tokodaii sp. nov. The type strain is strain 7 (JCM 10545). PMID- 11878561 TI - Thermostable Pyrococcus furiosus DNA ligase catalyzes the synthesis of (di)nucleoside polyphosphates. AB - DNA ligase from the hyperthermophilic marine archaeon Pyrococcus furiosus (Pfu DNA ligase) synthesizes adenosine 5'-tetraphosphate (p4A) and dinucleoside polyphosphates by displacement of the adenosine 5'-monophosphate (AMP) from the Pfu DNA ligase-AMP (E-AMP) complex with tripolyphosphate (P3), nucleoside triphosphates (NTP), or nucleoside diphosphates (NDP). The experiments were performed in the presence of 1-2 microM [alpha-32P]ATP and millimolar concentrations of NTP or NDP. Relative rates of synthesis (%) of the following adenosine(5')tetraphospho(5')nucleosides (Ap4N) were observed: Ap4guanosine (Ap4G) (from GTP, 100); Ap4deoxythymidine (Ap4dT) (from dTTP, 95); Ap4xanthosine (Ap4X) (from XTP, 94); Ap4deoxycytidine (Ap4dC) (from dCTP, 64); Ap4cytidine (Ap4C) (from CTP, 60); Ap4deoxyguanosine (Ap4dG) (from dGTP, 58); Ap4uridine (Ap4U) (from UTP, <3). The relative rate of synthesis (%) of adenosine(5')triphospho(5')nucleosides (Ap3N) were: Ap3guanosine (Ap3G) (from GDP, 100); Ap3xanthosine (Ap3X) (from XDP, 110); Ap3cytidine (Ap3C) (from CDP, 42); Ap3adenosine (Ap3A) (from ADP, <1). In general, the rate of synthesis of Ap4N was double that of the corresponding Ap3N. The enzyme presented optimum activity at a pH value of 7.2-7.5, in the presence of 4 mM Mg2+, and at 70 degrees C. The apparent Km values for ATP and GTP in the synthesis of Ap4G were about 0.001 and 0.4mM, respectively, lower values than those described for other DNA or RNA ligases. Pfu DNA ligase is used in the ligase chain reaction (LCR) and some of the reactions here reported [in particular the synthesis of Ap4adenosine (Ap4A)] could take place during the course of that reaction. PMID- 11878562 TI - Rhodothermus marinus: a thermophilic bacterium producing dimeric and hexameric citrate synthase isoenzymes. AB - Two separate citrate synthases from the extremely thermophilic bacterium Rhodothermus marinus have been identified and purified. One of the enzymes is a hexameric protein and is the first thermostable, hexameric citrate synthase to be isolated. The other is a dimeric enzyme, which is also thermostable but possesses both citrate synthase and 2-methyl citrate synthase activities. 2-Methyl citrate synthase uses propionyl-coenzyme A as one of its substrates and in Escherichia coli, for example, it has been implicated in the metabolism of propionate. However, no growth of R. marinus was observed using minimal medium with propionate as the sole carbon source, and both hexameric and dimeric enzymes were produced irrespective of whether propionate was included in the growth medium. The data are discussed with respect to the evolutionary relationships between the known hexameric and dimeric citrate synthases and 2-methyl citrate synthase. PMID- 11878563 TI - Characterization of Symbiobacterium toebii, an obligate commensal thermophile isolated from compost. AB - A symbiotic thermophilic bacterium, strain SC-1, was isolated from hay compost (toebi) in Korea. The new isolate exhibited an obligate commensal interaction with a thermophilic Geobacillus strain and required crude extracts and/or culture supernatant from Geobacillus sp. SK-1 for axenic growth. The growth factors from Geobacillus sp. SK-1 were irreversibly inactivated by phenol or protease treatment, suggesting that they might be proteins. The cells of strain SC-1 were non-spore forming, nonmotile rods that were stained Gram-negatively. The isolate was a microaerophilic heterotroph. Growth was observed between 45 degrees and 70 degrees C (optimum: 60 degrees C; 2.4-h doubling time) and pH 6.0 and 9.0 (optimum: pH 7.5). The G+C content of the genomic DNA was 65 mol%, and the major quinones were MK-6 and MK-7. A phylogenetic analysis of its 16S rDNA sequence indicated that strain SC-1 is closely related to Symbiobacterium thermophilum and so was named Symbiobacterium toebii on the basis of its physiological and molecular properties. PMID- 11878564 TI - A novel species of alkaliphilic Bacillus that produces an oxidatively stable alkaline serine protease. AB - A novel gram-positive, strictly aerobic, motile, sporulating, and facultatively alkaliphilic bacterium designated KSM-KP43 was isolated from a sample of soil. The results of 16S rRNA sequence analysis placed this bacterium in a cluster with Bacillus halmapalus. However, the level of the DNA-DNA hybridization of KSM-KP43 with B. halmapalus was less than 25%. Moreover, the G + C contents of the genomic DNA were 41.6 mol% for KSM-KP43 and 38.6 mol% for B. halmapalus. Because there were also differences in physiological properties and cellular fatty acid composition between the two organisms, we propose KSM-KP43 as a novel species of alkaliphilic Bacillus. This novel strain produces a new class of protease, an oxidatively stable serine protease that is suitable for use in bleach-based detergents. The enzyme contained 640 amino acid residues, including a possible approximately 200-amino-acid prepropeptide in the N-terminal and a unique stretch of approximately 160 amino acids in the C-terminal regions (434-amino-acid mature enzyme with a calculated molecular mass of 45,301 Da). The C-terminal half after the putative catalytic Ser255 and the contiguous C-terminal extension shared local similarity to internal segments of a membrane-associated serine protease of a marine microbial assemblage and the serine protease/ABC transporter precursors of the slime mold Dictyostelium discoideum, and to the C-terminal half of a cold active alkaline serine protease of a psychrotrophic Shewanella strain. PMID- 11878566 TI - Global communication is central to health. PMID- 11878565 TI - Identification, cloning, and expression of uracil-DNA glycosylase from Atlantic cod (Gadus morhua): characterization and homology modeling of the cold-active catalytic domain. AB - Two distinct forms of the highly conserved uracil-DNA glycosylase (UNG) have been isolated from Atlantic cod (Gadus morhua) liver cDNA by rapid amplification of cDNA ends (RACE). From the cDNA sequences, both forms were deduced to encode an open reading frame of 301 amino acids, with an identical 267-amino-acid C terminal region and different N-terminal regions of 34 amino acids. By comparison with the human UNG sequences, the two forms were identified as possible mitochondrial (cUNG1) and nuclear (cUNG2) forms. Several constructs of recombinant cUNG (rcUNG) were expressed in Escherichia coli in order to optimize the yield. The recombinant enzyme was purified to apparent homogeneity as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Activity and stability experiments showed that rcUNG was similar to cUNG previously purified from Atlantic cod liver, and was more pH- and temperature labile than a recombinant human UNG (rhUNG). Under optimal assay conditions for both rcUNG and rhUNG, the turnover number (k(cat)) was three times higher for rcUNG compared with rhUNG, with an identical K(M), resulting in a threefold higher catalytic efficiency (k(cat)/K(M)) for rcUNG. These activity and stability experiments reveal cold-adapted features in rcUNG. Homology models of the catalytic domains of Atlantic cod (cUNG) and mouse uracil-DNA glycosylase (mUNG) were built using the human UNG (hUNG) crystal structure as a template. The unique amino acid substitutions observed in cod UNG were mainly located in the N- and C-terminal parts of the sequence. The analysis indicated a more stable N-terminal, a more flexible C-terminal, and a less stabilized core in cUNG as compared with the mammalian UNGs. Substitution of several amino acids in or near the DNA-binding site in cUNG could give rise to a more positively charged surface and a higher electrostatic potential near the active site compared with the mammalian UNGs. The higher potential may increase the electrostatic interactions between the enzyme and DNA, and may explain the increased substrate affinity and, in combination with the higher flexibility, the higher catalytic efficiency observed for rcUNG. PMID- 11878567 TI - Dietary supplement safety information in magazines popular among older readers. AB - Dietary supplements are extensively used in the United States, especially by people age 50 and over. Surveys have shown that magazines and other news media are an important source of information about nutrition and dietary supplements for the American public. It is uncertain, however, whether magazines provide their readers with adequate information about the safety aspects of supplement use. This report presents an analysis of supplement safety information in articles published during 1994-1998 in 10 major magazines popular among older readers. This time period was chosen to allow the impact of the 1994 Dietary Supplement Health and Education Act (DSHEA) to be assessed. The evaluation included 254 magazine articles. More than two-thirds of the articles did not include comprehensive information about the safety aspects of the dietary supplements that were discussed. Information about safety issues such as maximum safe doses and drug-supplement interactions was often lacking even in otherwise informative and well-researched articles. A total of 2,983 advertisements for more than 130 different types of supplements were published in the magazines surveyed. The number of advertisements per year increased between 1995 and 1998. Supplements of particular interest to older adults (such as antioxidants, calcium, garlic, ginkgo biloba, joint health products, liquid oral supplements, and multivitamins) were among the most frequently advertised products. Although magazines popular among older readers contain extensive information about dietary supplements, these publications cannot be relied upon to provide readers with all of the information that they need in order to use supplements safely. PMID- 11878568 TI - Development of targeted message concepts for recent Asian immigrants about secondhand smoke. AB - Residents of Boston's Chinatown and the Vietnamese community in Boston's Dorchester section are recent immigrants from China and Vietnam, countries whose smoking prevalence rates for men are among the highest in the world and whose rates for women are very low. We conducted exploratory focus groups in these communities to examine issues related to secondhand smoke and to generate message concepts for health education materials that would convince recent Asian immigrants to respond to the public health threat that secondhand smoke poses. The message concepts, which were tailored specifically for Chinese and Vietnamese immigrants, used themes that were consistent with the cultural values of each group as expressed in the focus groups, yet also reflected the fact that, in many ways, these immigrants are seeking to adapt to American norms. We suggest that it is possible to construct culturally appropriate health education materials for recent immigrant populations rather than rely on simple translations of English language materials. An intervention study using these message concepts is needed. PMID- 11878569 TI - Youth performing arts entertainment-education for HIV/AIDS prevention and health promotion: practice and research. AB - Entertainment-education approaches to health promotion and disease prevention are a popular method for many interventions that target adolescents and young adults. This article documents how this approach is used to educate and influence young people about HIV/AIDS, other sexually transmitted diseases (STDs), and other health issues in the United States. A review of the literature is followed by a two-phase descriptive study of American youth performing arts entertainment education programs. First, a quantitative survey was conducted among youth performing arts participants who were attending a national conference on the subject. This was followed by a qualitative survey among adult and youth conference attendees from established HIV/AIDS prevention youth performing arts programs. These two approaches provided detailed insight into the characteristics, approaches, and frameworks used to create, implement, and evaluate these entertainment-education efforts. Nine domains that define the effects and effectiveness of youth HIV prevention entertainment-education interventions are identified and described, including those related to performances, intervention management, and audiences. Given the importance of evaluation for the success and effectiveness of intervention programs, these domains are used to construct a framework for entertainment-education research and evaluation efforts. PMID- 11878570 TI - The influence of organizational characteristics and campaign design elements on communication campaign quality: evidence from 91 Ugandan AIDS campaigns. AB - This research proposes and tests a model of the relationship between organizational factors, campaign design elements, and campaign quality of communication campaigns. It is the first quantitative study to test these relationships across many organizations. The context for the study was AIDS education and outreach campaigns in Uganda, during a time of successful decrease in the spread of HIV infection. Ninety-one organizations were surveyed. Since only 14% of the organizations collected exposure or outcome data, the study focused on the factors affecting campaign quality. Quality was examined by measuring goal specificity, execution quality, and message quality. The results show that financial resources, professional training, participation of outreach workers in planning the campaign, and audience participation in planning and executing the campaign were key organizational variables affecting the quality of the campaigns. The important campaign design elements affecting campaign quality were conducting research, using multiple channels, targeting only a few groups, and pretesting messages. The results have import for campaign planners, managers of organizations conducting campaigns, and funders. In addition, it is vital that organizations collect exposure and outcome data in the future to provide feedback on each campaign. PMID- 11878571 TI - Risk assessment in acute lymphoblastic leukemia: beyond leukemia cell characteristics. PMID- 11878572 TI - Blood money: ethical and legal implications of treating cord blood as property. PMID- 11878573 TI - Childhood acute lymphoblastic leukemia: prognostic value of initial peripheral blast count in good responders to prednisone. AB - PURPOSE: To assess the value of initial peripheral blast count in patients with acute lymphoblastic leukemia (ALL) and prednisone good response (PGR). PATIENTS AND METHODS: From January 1990 to December 1995, 403 consecutive patients with newly diagnosed ALL were enrolled in the authors' protocol 1-ALL90-BFM/HPG. Prednisone good response was defined as a blast count of less than 1,000/microL and a prednisone poor response (PPR) as a blast count of at least 1,000/microL, both in peripheral smears, after 7 days of oral prednisone (60 mg/m2 per day) and one intrathecal dose of methotrexate. In the PGR group, patients were divided into two subgroups: patients who had less than 1,000 blasts/microL at diagnosis and those with at least 1,000 blasts/microL at diagnosis. RESULTS: Three-hundred thirty-seven patients (90%) had PGR and 37 had (10%) PPR. At 5-year follow-up, event-free survival estimates were 67 +/- 3.8% and 38 +/- 8% for PGR and PPR, respectively (P = 0.0001). In the PGR group, 114 patients (34%) had an initial blast count of less than 1,000/microL and 223 (66%) had an initial blast count of at least 1,000/microL. The authors compared the clinical and laboratory characteristics of these subgroups at diagnosis and outcome and detected significant differences in white cell count, incidence of T immunophenotype, and presence of mediastinal or spleen enlargement. However, there were no differences in response to induction treatment, death in complete remission, relapses, or event-free survival probability. CONCLUSIONS: In the PGR group, regardless of the initial blast count, both subgroups had the same outcome. The PGR group with an initial blast count of at least 1,000/microL had significantly higher white cell counts. T markers, and mediastinal or spleen enlargement at diagnosis. Response to prednisone is a practical, inexpensive, and good prognostic factor in childhood ALL. PMID- 11878574 TI - Analysis of TP53 mutations in relapsed childhood acute lymphoblastic leukemia. AB - TP53 is the most commonly mutated gene in human cancer, but TP53 mutations are present in less than 5% of children with acute lymphoblastic leukemia (ALL) at initial presentation. Mutations are detected more frequently in children with relapsed T-cell ALL, but the potential role of TP53 mutations in relapsed B lineage childhood ALL is not understood as well. The authors determined the nucleotide sequence of amplified DNA from exons 5 to 8 of the TP53 gene in leukemic cells obtained from 17 children with ALL at the time of first bone marrow relapse. All 17 contained only germline TP53 sequences. Review of the published literature disclosed that TP53 mutations have been found in 22% of cases of relapsed ALL. To understand the role of p53 abnormalities in this clinical setting, it will be important for future studies to analyze cases of relapsed ALL with assays capable of interrogating the functional integrity of the p53 pathway. PMID- 11878575 TI - Evaluation of left ventricular function in asymptomatic children about to undergo anthracycline-based chemotherapy for acute leukemia: an outcome study. AB - BACKGROUND: Cardiac toxicity is a well-recognized potential complication of anthracycline use. Children treated with anthracyclines undergo several cardiac screening procedures before therapy, but the usefulness of these pretherapy cardiac studies has never been evaluated. The authors examined whether induction chemotherapy in patients with high-risk acute lymphoblastic leukemia (ALL) was altered based on a pretherapy left ventricular shortening fraction (SF). PATIENTS AND METHODS: Medical records of 134 children registered on treatment protocols of the Pediatric Oncology Group for high-risk B-precursor and T-cell ALL between 1987 and 1998 were reviewed. Demographic information consisting of age at diagnosis, sex, and past cardiac history was collected, as were the results of all echocardiographic evaluations for SF and actions taken based on these evaluations. The outcome measured was whether any changes were made in induction therapy based on initial SF. In addition, secondary SF results obtained at the cumulative anthracycline dose range of 90 to 150 mg/m2 were studied to determine whether modifications of future chemotherapy were made after this limited exposure. RESULTS: Three of 128 children (2.3%) without a previous cardiac history had an initial SF on their pretherapy echocardiogram that prompted additional evaluation but no change in therapy. A secondary analysis of SF in 85 children who completed anthracycline doses of 90 to 150 mg/m2 was performed. There were three (3.5%) with abnormal study results who were evaluated further. Again, no changes were made in the anthracycline doses based on these findings. No cardiac dysfunction occurred among these six patients during later follow-up. CONCLUSIONS: In the absence of a previous cardiac history or signs and symptoms or cardiac disease, pretherapy evaluation of left ventricular function may not be indicated in children about to undergo anthracycline-based treatment of acute leukemia. The timing of initiation of cardiac evaluation remains unclear, but these results suggest that even at a cumulative dose of 90 to 150 mg/m2, studies to determine left ventricular function do not yield data sufficient to warrant a change in the clinical management of these patients. PMID- 11878576 TI - Cardiovascular risk factors in young adult survivors of childhood acute lymphoblastic leukemia. AB - PURPOSE: To assess cardiovascular risk factors (CVRF) in young adult survivors of childhood acute lymphoblastic leukemia (ALL). PATIENTS AND METHODS: Twenty-six subjects (median age, 20.9 years; median interval since completion of therapy, 13.3 years) were evaluated. Ten participants had received cranial irradiation (CRT), whereas 16 had received only chemotherapy. Primary outcome measures included body mass index (BMI), blood pressure, fasting lipoprotein, glucose, and insulin levels. Secondary measures included insulin-like growth factor-1 (IGF-1) and IGF binding protein-3 levels, physical activity index, a 7-day dietary recall, tobacco product use, and measurement of the intima-media thickness (IMT) of the common carotid artery. RESULTS: Sixty-two percent (16/26) of participants had at least one CVRF potentially related to their cancer treatment (obesity, dyslipidemia, increased blood pressure, or insulin resistance), with 30% (7/26) having more than two CVRF. Thirty-one percent (8/26) of subjects were obese (BMI > or = 30). Subjects who were treated with CRT (BMI, 30.4 +/- 6.7) had an increased BMI (P = 0.039) in comparison with those who received only chemotherapy (BMI, 25.4 +/- 5.1). Triglyceride and very low-density lipoprotein C levels were significantly higher in those treated with CRT (P = 0.027 and 0.022, respectively). The IGF-1 was inversely correlated with IMT (total group, -0.514, P = 0.009; females only, -0.729, P = 0.003). CONCLUSIONS: Young adult survivors of childhood ALL, especially those treated with CRT, are at risk for obesity and dyslipidemia, insulin resistance, hypertension, and cardiovascular disease. Further investigation of these risks is warranted. PMID- 11878577 TI - Prognostic factors and secondary malignancies in childhood medulloblastoma. AB - PURPOSE: Little is known of the outcome of long-term survivors of childhood medulloblastoma, one of the most common pediatric malignancies. To determine the potential for secondary malignancies, a retrospective outcome evaluation in 88 consecutive cases of childhood medulloblastoma was performed. PATIENTS AND METHODS: The records of all patients with childhood medulloblastoma diagnosed at Children's National Medical Center in Washington, DC from 1969 through 1997 were reviewed. RESULTS: The median follow-up time was 92 months (range 6-257 months). Overall survival was 59% at 5 years and 52% at 10 years. Univariate analysis showed that age at diagnosis, extent of surgical resection, presence of metastatic disease (M stage), ventriculoperitoneal shunt placement within 30 days from diagnosis, posterior fossa radiation therapy dose, and adjuvant chemotherapy significantly affected survival. Although based on small numbers, the risk of second neoplasms was significantly increased in this cohort. Multiple basal cell carcinomas developed in the areas of radiation therapy in two patients; these patients also had nevoid basal cell carcinoma syndrome (NBCCS) diagnosed. One other patient died of glioblastoma multiforme 8 years after treatment of medulloblastoma. A meningioma developed in another patient 10 years after radiation therapy. CONCLUSION: As survival of medulloblastoma patients improves, increased surveillance regarding secondary malignancies is required, especially because radiation-induced tumors may occur many years after treatment. These two cases of NBCCS also illustrate the importance of considering the concomitant diagnosis of NBCCS in young patients with medulloblastoma. In those patients, alternative therapy should be considered to minimize radiation therapy-related sequelae. PMID- 11878578 TI - Follow-up in neuroblastoma: comparison of metaiodobenzylguanidine and a chimeric anti-GD2 antibody for detection of tumor relapse and therapy response. AB - Early and correct diagnosis of local tumor recurrence, occurrence of metastases, and therapy response are essential in patients with neuroblastoma stage IV. The aim of the study was to evaluate the diagnostic value of metaiodobenzylguanidine (mIBG) and a chimeric GD2 antibody in the follow-up of patients with neuroblastoma. In a prospective study, mIBG (N = 31 scans) and immunoscintigraphy were compared with a chimeric antiganglioside antibody, ch14.18 (MAb) (N = 31 scans), labeled with technetium Tc 99m in the follow-up of 18 patients with stage IV neuroblastoma. The findings were compared with histologic findings, other imaging examinations, and clinical changes over the course of 4 to 6 years. For the diagnosis of local tumor recurrences, sensitivity was 80% for MAb and 70% for mIBG. Specificity was 93% for MAb and 72% for mIBG. The MAb was superior for the detection of skeletal metastases, with a sensitivity of 82% compared with 72% for mIBG. Specificity was 100% for both techniques. Also, for soft tissue/lymph node metastases, sensitivity for MAb was higher (50%) than for mIBG (31%). Specificity was 100% for each technique. In sequential studies, metastases were detected earlier with MAb (mean: 2.3 m for skeletal metastases, 3.6 m for soft tissue metastases) than with mIBG. After therapy, tumor uptake was visualized longer with mIBG (mean 6.3 m) than with MAb. The chimeric antibody ch14.18 is likely to be valuable for follow-up examinations and for assessment of therapy response because of earlier detection of new metastases. PMID- 11878579 TI - Deep soft tissue infections in the neutropenic pediatric oncology patient. AB - PURPOSE: Necrotizing fasciitis and myonecrosis can be rapidly fatal without prompt and aggressive medical and surgical therapy. We reviewed our experience with necrotizing fasciitis and myonecrosis in neutropenic pediatric oncology patients to describe associated clinical characteristics and outline therapeutic interventions. PATIENTS AND METHODS: A retrospective chart review was performed for all cases of deep soft tissue infection found in neutropenic pediatric oncology patients during an 11-year period. RESULTS: Seven cases of necrotizing fasciitis and/or myonecrosis associated with chemotherapy-induced neutropenia were diagnosed during the study period. Deep soft tissue infection was diagnosed a median of 14 days after the initiation of chemotherapy. All of the patients presented with fever and pain, generally out of proportion to associated physical findings. Most patients (86%) also had tachycardia and subtle induration at the site of soft tissue infection. The pathogenic organism in four of seven patients originated in the gastrointestinal tract. Patients were treated with antibiotics, surgical debridements, granulocyte colony-stimulating factor, and hyperbaric oxygen. Granulocyte transfusions were administered if there were no signs of neutrophil recovery. Five patients survived their deep soft tissue infection. CONCLUSIONS: The diagnosis of necrotizing fasciitis and/or myonecrosis should be considered in any neutropenic patient with fever, tachycardia, and localized pain out of proportion to the physical findings. Appropriate therapy includes broad spectrum intravenous antibiotics and urgent surgical intervention. Granulocyte colony-stimulating factor should be administered to all patients to enhance neutrophil recovery. Granulocyte transfusions should be considered if a prolonged period of neutropenia is anticipated. PMID- 11878580 TI - Influence of bilirubin uridine diphosphate-glucuronosyltransferase 1A promoter polymorphisms on serum bilirubin levels and cholelithiasis in children with sickle cell anemia. AB - PURPOSE: Genetic mutations in the uridine diphosphate (UDP) glucuronosyltransferase 1A (UGT1A) enzyme promoter have been associated with unconjugated hyperbilirubinemia and Gilbert syndrome. The effects of UGT1A promoter polymorphisms on serum bilirubin levels and symptomatic gallstone formation were studied in a cohort of children with sickle cell anemia (SCA). METHODS: The UGT1A promoter genotype was deterrmined for 115 consecutive children with SCA. Steady-state laboratory parameters and previous cholecystectomy for symptomatic gallstones were recorded retrospectively, then analyzed according to UGT1A genotype. RESULTS: Children with SCA had a lower frequency of the normal (TA)6 UGT1A promoter allele (0.413) than the abnormal (TA)7 allele (0.461). A previously described shorter (TA)5 allele (frequency 0.074) and longer (TA)8 allele (frequency 0.052) were also observed. Children with the 7/7 UGT1A genotype had a significantly higher mean bilirubin level (5.8 +/- 3.1 mg/dL) than those with the 6/6 (2.4 +/- 0.8 mg/dL) or 6/7 genotype (3.0 +/- 1.1 mg/dL; P < 0.001 by analysis of variance). Patients with the 7/7 genotype were more likely to have previous cholecystectomy (87.5%) than those with the 6/6 (35.7%) or the 6/7 genotype (36.1%; P = 0.002 by chi2). CONCLUSIONS: Genetic variation in the UGT1A promoter significantly influences serum bilirubin levels and the development of symptomatic cholelithiasis in children with SCA. The UGT1A promoter polymorphisms represent an important nonglobin genetic modifier of clinical disease expression in SCA. PMID- 11878581 TI - Modified cyclophosphamide, hydroxydaunorubicin, vincristine, and prednisone therapy for posttransplantation lymphoproliferative disease in pediatric patients undergoing solid organ transplantation. AB - PURPOSE: The authors report the use of a cyclophosphamide, hydroxydaunorubicin, vincristine, and prednisone (CHOP)-based chemotherapy regimen in treating six children with posttransplantation lymphoproliferative disorder (PTLD) that developed after solid organ transplantation. MATERIALS AND METHODS: The chemotherapy regimen consisted of a 29-day induction with CHOP and then as many as 15 cycles of maintenance therapy using methotrexate and cytarabine alternating with vincristine, adriamycin, mercaptopurine, and prednisone. RESULTS: All patients attained remission. One patient died of sepsis while in remission. Four of the five remaining patients have been followed-up in remission for as long as 8 years without losing the graft. One of the patients experienced relapse after completing therapy and subsequently died with disease. CONCLUSIONS: The authors conclude that pediatric patients with PTLD after solid organ transplantation that fails conservative management can be treated successfully with CHOP-based chemotherapy. PMID- 11878582 TI - Fatal myocardial aspergillosis in an immunosuppressed child. AB - A girl with resistant acute myeloid leukemia (AML) had a stem cell transplantation. Preceding transplantation, she had recurrent pneumonitis. No causative agent was identified. Despite several antibiotics including high-dose liposomal amphotericin-B, pulmonary infection progressed. Aspergillosis, always considered, could not be documented. She died from cardiac arrest on the second day after transplantation, with no forewarning of previous heart disease. Pericardial and myocardial aspergillosis was an autopsy finding. Pericardial and myocardial aspergillosis, rare manifestations of systemic aspergillosis, should be considered in any immunocompromised patient with long-lasting pulmonary infection, even in the absence of specific cardiac findings. PMID- 11878583 TI - Intravenous anti-D immune globulin-induced intravascular hemolysis in Epstein Barr virus-related thrombocytopenia. AB - RhoD immune globulin intravenous (anti-D IGIV) increases platelet counts in patients who have not undergone splenectomy and are positive for RhoD with idiopathic thrombocytopenic purpura. After treatment, in most patients, anemia develops as a result of immune-mediated red cell destruction in the spleen. Although intravascular hemolysis (IVH) is not expected, life-threatening IVH has been recently reported by the Food and Drug Administration, and physicians are encouraged to report their experience with patients with idiopathic thrombocytopenic purpura in whom IVH develops after anti-D administration. Severe IVH was observed after treatment with anti-D IGIV in two adolescent girls with acute thrombocytopenia related to Epstein-Barr virus. They did not have hemolytic anemia before treatment. The authors believe that anti-D IGIV triggered an unusual virus-induced immune response causing hemolysis; therefore, anti-D IGIV should not be used in patients with Epstein-Barr virus-related thrombocytopenia, particularly during the acute phase of infection. PMID- 11878584 TI - Griscelli syndrome: rare neonatal syndrome of recurrent hemophagocytosis. AB - Griscelli syndrome (GS) is a rare inherited disease characterized by immunodeficiency and partial albinism. The microscopic findings of the skin and hair are highly suggestive of the disease. The GS locus colocalizes on chromosome 15q21 with the myosin-Va gene (MYO5a), and mutations have been identified in few patients. We describe a 2-month-old Hispanic girl with severe pancytopenia secondary to hemophagocytosis. Even though a mutation at the Griscelli locus had not been identified, her clinical features and outcome were typical of GS. The purpose of this article is to alert physicians to the association between GS and hemophagocytosis. We suggest that GS should be considered in infants with hemophagocytosis because the features of partial albinism can be subtle. The relevant literature is summarized. PMID- 11878585 TI - Thrombocytopenia absent corpus callosum syndrome: third case of a distinct clinical entity. AB - Thrombocytopenia absent corpus callosum, characterized by refractory thrombocytopenia, agenesis of the corpus callosum, hypoplastic cerebellum, abnormal facies, and developmental delay, represents a relatively newly described clinical entity. An 18-month-old girl with agenesis of the corpus callosum, hypoplasia of the cerebellar vermis, hypotonia, and severe developmental delay presented with thrombocytopenia. She had a distinctive facies with microcephaly, broad nasal root with upturned nose, small upper lip, and micrognathia. A bone marrow aspirate and biopsy showed normal cellularity with dysplastic megakaryocytes. Thrombocytopenia absent corpus callosum is compared with other conditions characterized by congenital non-immune thrombocytopenia. PMID- 11878586 TI - Unusual delayed complication of central venous access. PMID- 11878587 TI - Quantitation of nicotine in tobacco products by capillary electrophoresis. AB - A simple and rapid capillary electrophoresis (CE) method was developed for the quantitation of nicotine in commercial tobacco products. The method involves a 6 min run at 30 kV, using a 50 mM phosphate buffer (pH 2.5), paraquat as internal standard, and UV detection at 260 nm. Nicotine was extracted from tobacco products in <15 min. Recoveries from spiked extracts were >95%, and the extraction efficiencies of water, 1 M HCI, 1 M acetic acid, 5 mM phosphate buffer (pH 2.5), and 1% triethanol amine were similar. Nicotine concentrations in 67 samples of cigarettes, cigars, and bidis varied between 0.37 and 2.96% (w/w). An established gas chromatography/mass spectrometry method using toluene extraction consistently yielded lower nicotine values than the CE method. Experimental evidence suggests that this is due to insufficient extraction of nicotine by toluene. PMID- 11878588 TI - Evaluation of the use of liquid dishwashing compounds to control bacteria in kitchen sponges. AB - A test procedure for evaluating the effect of adding commercial liquid hand dishwashing detergents to kitchen sponges to control microbial growth is described. Claims for this type of application are being made on dishwashing detergents throughout the world. In this evaluation, commercially available kitchen sponges were stripped of antimicrobial compounds. Sponges were then inoculated with a pool of 7 microorganisms which consisted of gram positives, gram negatives, and yeast. Inoculated sponges were treated with the detergent as recommended by the manufacturer and allowed to incubate for 16 h at ambient temperature. Surviving microorganisms were then quantitated using either the spiral or pour plate method. Tests were run using both clean sponges and sponges soiled with 0.5% nonfat dry milk (NFDM). Untreated sponges showed stasis or slightly increased bacterial populations after the incubation period in the absence of NFDM. Significant increases of up to 3 log cfu/mL were observed for untreated sponges when soiled with NFDM. Statistically significant reductions were observed for clean sponges (99.8-99.9998%) and sponges soiled with NFDM (87.6-99.9%) when detergents making "antibacterial sponge" claims were added to the inoculated sponges. Statistically significant differences between detergents making "antibacterial sponge" claims were also observed. PMID- 11878589 TI - Multielemental analysis of food and agricultural matrixes by inductively coupled plasma-mass spectrometry. AB - The suitability of quadrupole inductively coupled plasma-mass spectrometry (ICP MS) for multielemental analysis of food following microwave closed vessel digestion of samples was evaluated in relation to analytical challenges presented by some major food and agricultural matrixes. Fifteen key analytes (Al, As, Cd, Co, Cr, Cu, Fe, Mn, Mo, Ni, Pb, Se, Sn, V, Zn) were determined in 9 reference materials representative of 3 major groups of staple foods (cereals, seafood, and meat). For all measurements, the method of external calibration was used and Rh was selected as internal standard. Matrix-induced interferences were evaluated for each material, and suitable methods to overcome them were applied. Spectral interferences caused by C, Cl, and Ca content of digestates were quantitated and corrected by entering a simple system of mathematical correction equations into the instrument software before each analytical run. Full mastering of interferences together with adoption of a series of measures to control the reliability of analytical measurements produced accurate results for all the analytes. A standard analytical protocol is outlined for the benefit of both research and routine high-throughput laboratories that perform ICP-MS analyses of food. PMID- 11878590 TI - Optimization and validation of arsenic determination in foods by hydride generation flame atomic absorption spectrometry. AB - A hydride generation flame atomic absorption spectrometric method was developed and optimized to quantitate arsenic (As) in foods. A wet digestion of the samples with HNO3 + H2O2 was performed and excess oxidants were eliminated by addition of hydrochloric acid and urea. As5+ in As3+ was then reduced by potassium iodide. The As3+ solution was analyzed by generation of arsine with sodium tetrahydroborate. As determination ranged from 2.5 to 20 microg/L, with a determination coefficient of 0.997. The limits of detection (LOD) and quantitation (LOQ) were 0.6 and 2.1 microg/L, respectively. The method was validated and good results were obtained for recovery, precision, accuracy, LOD, and LOQ. This method is now used to analyze foods from Rwanda. PMID- 11878591 TI - Determination of insecticides in honey by matrix solid-phase dispersion and gas chromatography with nitrogen-phosphorus detection and mass spectrometric confirmation. AB - A multiresidue method was developed for the determination of 12 organophosphorus insecticides (diazinon, parathion methyl, fenitrothion, pirimiphosmethyl, malathion, fenthion, chlorpyrifos, quinalphos, methidathion, ethion, azinphosmethyl, coumaphos), one carbamate (pirimicarb), and one amidine (amitraz) in unifloral and multifloral honeys. The analytical procedure was based on the matrix solid-phase dispersion of honey on a mixture of Florisil and anhydrous sodium sulfate in small glass columns and subsequent extraction with a low volume of hexane-ethyl acetate (90 + 10, v/v), assisted by sonication. The insecticide residues were determined by capillary chromatography with nitrogen-phosphorus detection and confirmed by mass spectrometry. Average recoveries at the 0.05-0.5 microg/g levels were >80% for organophosphorus insecticides and about 60% for the other insecticides, pirimicarb and amitraz, with relative standard deviations <10%. The detection limit for the different insecticides ranged between 6 and 15 microg/kg. The main advantages of the proposed method are that extraction and cleanup are performed in a single step with a low volume of organic solvent. The method is simple, rapid, and less laborious than conventional methods. Several Spanish honeys were analyzed with the proposed method and no residues of the studied insecticides were found. PMID- 11878592 TI - Dietary exposure of Thais to pesticides during 1989-1996. AB - To monitor the exposure of the Thai to pesticide residues in foods, the intake of pesticides based on the total diet approach has been studied in Thailand continually since 1989. Food items were chosen and their proportions of daily consumption were calculated from the national food consumption survey conducted by the Department of Health in 1986. Seventy-seven items of food including drinking water were classified into 12 groups according to their sources of contamination and the analytical methods used. Shopping lists and standard procedures for collecting samples and cooking were developed. Twelve food composites from each of the 4 regions were analyzed for nearly 100 pesticides. All analyses used multiresidue analytical methods, and the total daily dietary intakes of pesticide residues were estimated from the average amount of food consumed by Thais. Among 24 pesticides found in the 8 year study, DDT, dimethoate, methamidophos, and parathion methyl were found every year. However, dietary intakes of all pesticides were far below the established acceptable daily intake. In 1999, the study methodology was redesigned to accommodate the new national food consumption survey data. PMID- 11878593 TI - Determination of polycyclic aromatic hydrocarbons from buzzards (Buteo buteo) and tawny owl (Strix aluco) by liquid chromatography with fluorescence detection. AB - Supercritical fluid extraction was applied to the determination of naturally contaminated polycyclic aromatic hydrocarbons (PAHs) in bird tissue by liquid chromatography with fluorescence detection (LC-FL). Recoveries (> 90%) and relative standard deviations (< or = 7.7%) were satisfactory. The levels of 10 PAHs were analyzed in 6 classes of tissues (heart, liver, intestine, muscle, lung, and kidney) of 10 buzzards and 2 tawny owls, predatory birds from the Galicia (northwest Spain). The PAHs found most abundantly were pyrene, fluoranthene, benzo[a]anthracene, and anthracene. Chrysene, benzo[b]fluoranthene, benzo[k]fluoranthene, benzo[ghi]perylene, and indeno[1,2,3-cd]pyrene were not detected. Intestine, kidney, and lung were more polluted than other tissues. PMID- 11878594 TI - Rapid food decomposition by H2O2-H2SO4 for determination of total mercury by flow injection cold vapor atomic absorption spectrometry. AB - A mixture of 50% H2O2-H2SO4 (3 + 1, v/v) was used for decomposition of food in open vessels at 80 degrees C. The treatment allowed rapid total mercury determination by flow injection cold vapor atomic absorption spectrometry. Cabbage, potatoes, peanuts paste, hazelnuts paste, oats, tomatoes and their derivatives, oysters, shrimps, prawns, shellfish, marine algae, and many kinds of fish were analyzed by the proposed methodology with a limit of quantitation of 0.86 +/- 0.08 microg/L mercury in the final solution. Reference materials tested also gave excellent recovery. PMID- 11878595 TI - Liquid chromatographic determination of amphotericin B in different pharmaceuticals. AB - Amphotericin B (AmB) is one of the most potent antifungal agents and the drug of choice in the treatment of serious fungal infections. A liquid chromatographic (LC) method was developed to determine AmB in pharmaceutical formulations for injection, tissue culture, cream, and lotion. pBondapak C18 reversed-phase column and a simple mobile phase consisting of acetonitrile-water-acetic acid (40 + 54 + 6, v/v) was used. The flow rate was 1.8 mL/min and the effluent was monitored at 405 nm. The developed LC method uses piroxicam as an internal standard and has a limit of detection of 10 ng/mL, a limit of quantitation of 30 ng/mL, and the assay is linear from 0.01 to 100 microg/mL. AmB and piroxicam elute with retention times of 12.4 and 4.0 min, respectively, and the resolution between AmB and piroxicam was 10.6. In comparison with the official United States Pharmacopeia microbial assay for AmB, this LC method is more rapid, selective, sensitive, and offers positive identification. PMID- 11878596 TI - A review of analytical methods for the determination of sulfolane and alkanolamines in environmental studies. AB - Sulfolane and alkanolamines are used extensively in the processing of sour natural gases. Over many years of operation, there have been inadvertent leaks of these chemicals to groundwater and wetlands surrounding gas processing facilities, leading to uptake by vegetation. Because sulfolane and alkanolamines are extremely water-soluble, their analysis has presented challenges, particularly requirements for suitable extraction from biological matrixes and soil, along with sensitive detection using commonly available instrumentation. Analytical methods usually use gas chromatography or liquid chromatography with a variety of detector systems. Sample preparation techniques may include extraction with organic solvents, water, or a combination of these. In some cases, direct aqueous injections have been used. Derivatization of alkanolamines has been used to improve the chromatographic separations and detection. More recent procedures, using positive-ion electrospray ionization mass spectrometry (MS), have been useful for the confirmation of uptake of the alkanolamines and transformation products by wetland vegetation. Future developments will likely center on further MS analyses for identification of metabolites and transformation products in aquatic environments. PMID- 11878597 TI - Development of the infrared hollow waveguide sampler for the detection of chlorophenols in aqueous solutions. AB - A method based on the infrared hollow waveguide sampler was developed for sensing chlorophenols in aqueous solutions. This sampler was constructed by coating a suitable hydrophobic film onto the inner surface of an infrared hollow waveguide. By passing the aqueous solution through the hollow waveguide sampler, analytes can be absorbed into the hydrophobic layer. The adsorbed analytes can be sensed later by using Fourier transform infrared spectrometry. Six hydrophobic polymers were investigated for their performance in conjunction with the infrared hollow waveguide sampler for the detection of chlorophenols. Results indicated that poly(acrylonitrile-co-butadiene) was a most suitable hydrophobic material for absorption of chlorophenols in aqueous solutions. To further increase the detection sensitivity, factors such as sampling flow rate, sampling time, and thickness of the hydrophobic film were also investigated. Results indicated that the infrared signals were similar in the examined flow rates (2-30 mL/min), but that a higher flow rate tended to produce a higher analytical signal. Fast detection speed was an advantage of this method for the detection of chlorophenols, and the sampling/detection time can be <10 min. In addition, analytical signals were nearly proportional to the thickness of the hydrophobic film coating the inside of the hollow waveguide. With the optimal conditions found in this work, detection limits based on 3 times the peak-to-peak noise level were around 300 ppb for the chlorophenols examined. A high degree of linearity in the standard curves was also observed for this method in the concentration range of 10-100 ppm. The typical regression coefficients were >0.994 for the chlorophenols examined. PMID- 11878598 TI - Evaluation of the mixed aggregate method as an alternative to the bismuth active substances and cobalt thiocyanate active substances procedures for the determination of nonionic surfactants in raw and treated sewage. AB - The mixed aggregate (MA) method was evaluated for the routine determination of total ethoxylated nonionic surfactants in municipal raw and treated sewage. Nonionics in wastewater samples were enriched with a C8 solid-phase extraction cartridge, eluted with ethyl acetate-methanol, and separated from interferences with stacked strong anionic (quaternary ammonium form) and cationic (benzenesulfonic acid form) exchange columns. Sample extracts were evaporated to dryness and redissolved in distilled water. The total amount of ethoxylates, on a weight basis, was determined by measuring the critical micelle concentration of a mixed micelle made up of ethoxylates and the anionic surfactant sodium dodecyl sulfate, used as the reagent. The dye Coomassie Brilliant Blue G was used as a photometric probe. The minimum amount of ethoxylate required for a single determination was 1 microg. For raw sewage samples, mean recoveries and standard deviations were 98 +/- 6 and 98 +/- 4% at fortification levels of 0.3 and 0.6 microg/mL, respectively. For treated sewage samples, mean recoveries and standard deviations were 95 +/- 2 and 100 +/- 2% at fortification levels of 0.03 and 0.06 microg/mL, respectively. The MA method and the classical bismuth active substances and cobalt thiocyanate active substances procedures were compared. The former offers some significant advantages over the classical procedures in terms of sensitivity, selectivity, precision, simplicity, and rapidity; the type of response is independent of both the molecular weight and the chain length of the surfactant. PMID- 11878599 TI - Determination of dissolved naphthenic acids in natural waters by using negative ion electrospray mass spectrometry. AB - Naphthenic acids (NAs) have been implicated as some of the most toxic substances in oil sands leachates and identified as priority substances impacting on aquatic environments. As a group of compounds, NAs are not well characterized and comprise a large group of saturated aliphatic and alicyclic carboxylic acids found in hydrocarbon deposits (petroleum, oil sands bitumen, and crude oils). Described is an analytical method using negative-ion electrospray ionization mass spectrometry (ES/MS) of extracts. Preconcentration was achieved by using a solid phase extraction procedure utilizing a crosslinked polystyrene-based polymer with acetonitrile elution. Recovery of the Fluka Chemicals NA mixture was highly pH dependent, with 100% recovery at pH 3.0, but only 66 and 51% recoveries at pHs 7 and 9, respectively. The dissolved phase of the NA was very dependent on sample pH. It is thus critical to measure the pH and determine the appropriate mass profiles to identify NAs in natural waters. The ES/MS analytical procedure proved to be a fast and sensitive method for the recovery and detection of NAs in natural waters, with a detection limit of 0.01 mg/L. PMID- 11878600 TI - Simultaneous liquid chromatographic determination of 39 polycyclic aromatic hydrocarbons in indoor and outdoor air and application to a survey on indoor air pollution in Fuji, Japan. AB - An analytical method was established for the simultaneous determination of 39 polycyclic aromatic hydrocarbons (PAHs) in air. The method was applied to a survey of gaseous and particulate PAHs in household indoor air. The survey was performed in 21 houses in the summer of 1999 and in 20 houses in the winter of 1999-2000 in Fuji, Japan. Thirty-eight PAHs were determined in indoor and outdoor air in the summer, and 39 PAHs were determined in indoor and outdoor air in the winter. The concentrations of gaseous PAHs in indoor air tended to be higher than those in outdoor air in the summer and winter. The concentrations of particulate PAHs in indoor air were the same as or lower than those in outdoor air in the summer and winter. PAH profiles, correlations between PAH concentrations, and multiple regression analysis were used to determine the factors affecting the indoor PAH concentrations. These results showed that gaseous PAHs in indoor air were primarily from indoor emission sources, especially during the summer, and that indoor particulate PAH concentrations were significantly influenced by outdoor air pollution. PMID- 11878601 TI - Liquid chromatographic determination of multiple sulfonamides, nitrofurans, and chloramphenicol residues in pasteurized milk. AB - A rapid and selective liquid chromatographic method was developed to detect 6 sulfonamides, 3 nitrofurans, and chloramphenicol residues in pasteurized milk. The 10 drugs were extracted with chloroform-acetone and the organic phase was evaporated; the residues were dissolved in an aqueous sodium acetate buffer solution 0.02M (pH = 4.8), and the fat was removed by washing with hexane. The aqueous layer was collected, filtered, and injected. The 6 sulfonamides and chloramphenicol were detected at 275 nm ultraviolet (UV) using a gradient system starting with sodium acetate buffer solution-acetonitrile (95 + 5) and finishing with sodium acetate buffer solution-acetonitrile (80 + 20). Nitrofurans were detected at 375 nm (UV) isocratically with sodium acetate buffer solution acetonitrile (80 + 20). For 50 ppb fortified milk, the average recoveries were (sulfathiazole) 65.52%; (sulfamerazine) 75.36%; (sulfamethazine) 93.94%; (sulfachlorpyridazine) 75.94%; (sulfamethoxazole) 85.18%; (sulfamonomethoxine) 83.45%; (chloramphenicol) 104.17%; (nitrofurazone) 91.81%; (furazolidone) 100.76%; and (furaltadone) 72.38%. Method detection limits ranged from 4 ppb (nitrofurazone) to 16 ppb (sulfamethazine). Some matrix interferences (3-7 ppb) were observed only with sulfonamides. PMID- 11878602 TI - Gas chromatographic/mass spectrometric determination of benzene and its alkyl derivatives in indoor and outdoor air in Fuji, Japan. AB - An analytical method was established for the determination of benzene and 13 of its alkyl derivatives. The method was applied to a survey of indoor pollution that investigated the usefulness of the method, concentration levels, seasonal variations, profiles, correlations between compounds, and factors that affected indoor pollution by these compounds. The survey was performed in 21 houses in the summer of 1999 and 20 houses in the winter of 1999-2000 in Fuji, Japan. All the target compounds were detected in the indoor and outdoor air of all houses. Outdoor concentrations of benzene ranged from 0.779 to 3.17 microg/m3 in summer and from 1.35 to 6.04 microg/m3 in winter, whereas indoor concentrations of benzene ranged from 0.694 to 3.11 microg/m3 in summer and from 1.65 to 6.89 microg/m3 in winter. Indoor concentrations of the target compounds, except for benzene, were elevated, compared with outdoor concentrations. Because indoor and outdoor concentrations of benzene and its derivatives in summer were lower than in winter, the emission of these compounds may be increased by use of a heater and other variables present in winter. Profiles of the compounds, correlations between the compounds, and factors that affected indoor pollution (determined by multiple regression analysis) were investigated. These results suggested that indoor benzene predominantly penetrated from outdoors and that other benzene derivatives were emitted from indoor sources, such as paint solvents and kerosene heaters. PMID- 11878603 TI - Evaluation of Platanus occidentalis and Pinus sylvestris as bioindicators for lead and cadmium by slurry sampling-electrothermal atomic absorption spectrometry. AB - Two higher plants (Platanus occidentalis and Pinus sylvestris) were used as bioindicators for assessment of contamination by Pb and Cd in an urban and industrialized area. Both toxic metals in plant tissue were determined by transverse-heated electrothermal atomic absorption spectrometry-longitudinal Zeeman background correction combined with automated ultrasonic slurry sampling. Twenty sampling points were established to observe a relationship between metal concentration in plant and traffic density. Screening optimization of variables influencing slurry preparations (sonication time, sonication power, and surfactant concentration) followed a fractional factorial design at 2 levels. No significant effects were observed in the interval of variables studied. Limits of detection of Pb and Cd in the plants were 0.28 and 0.025 microg/g (dry weight), respectively. Metal concentrations in the plants were in the range of 5-51 and 0.05-0.7 microg/g for Pb and Cd, respectively. Factor analysis to data of metal concentration in both plants, extractable metal content in soil, distance to traffic, and traffic density, showed a significant association between features such as concentration of Pb and Cd in leaves of P. occidentalis and traffic density, which accounted for the largest variance. Other features such as extractable metal contents in soil accounted for lesser variance, meaning that soils in the area investigated were less suitable than plants for monitoring metal pollution caused by vehicular traffic. PMID- 11878604 TI - Study of the versatility of a graphite furnace atomic absorption spectrometric method for the determination of cadmium in the environmental field. AB - Cadmium is a representative example of trace elements that are insidious and widespread health hazards. In contemporary environmental analysis, there is a clear trend toward its determination over a wide range of concentrations in complex matrixes. This paper describes a versatile method for the determination of Cd at various levels (0.1-500 microg/g) in several sample types, such as soils, sediments, coals, ashes, sewage sludges, animal tissues, and plants, by graphite furnace atomic absorption spectrometry with Zeeman background correction. The effect of the individual presence of about 50 elements, with an interference/analyte concentration ratio of up to 10(5), was tested; recoveries of Cd ranged from 93 to 106%. The influence of different media, such as HNO3, HCI, HF, H2SO4, HClO4, acetic acid, hydroxylammonium chloride, and ammonium acetate, in several concentrations, was also tested. From these studies it can be concluded that the analytical procedure is scarcely matrix dependent, and the results obtained for a wide diversity of reference materials are in good agreement with the certified values. PMID- 11878606 TI - Employment of peak area as instrumental datum for improving the voltammetric quantitative analysis of heavy metals in fresh and sea water in the presence of surfactants. AB - Peak area instead of peak height is used for the simultaneous determination of Cu(II), Pb(II), Cd(II), and Zn(II) in fresh and sea water in the presence of anionic and cationic surfactants by differential pulse anodic stripping voltammetry. These species, if present, tend to make more irreversible the electrodic process of the metals. Because the employment of peak area for species having irreversible electrodic processes permits limits of detection about one or 2 orders of magnitude lower, this work shows the possibility of determining heavy metals at ultratrace level concentrations in the presence of surfactants, compounds which are always present in natural waters. The precision and accuracy of the analytical method were checked by the analysis of the standard reference materials (SRM) Fresh Water NIST-SRM 1643d, Sea Water BCR-CRM 403, and Estuarine Water BCR-CRM 505. The former, expressed as relative standard deviation (s(r)), and the latter, expressed as relative error (e), were satisfactory, being in all cases lower than 5%. The analytical procedure has been applied to fresh and sea water sampled in the Po river mouth area (Italy). PMID- 11878605 TI - Atomic absorption spectrometric determination of chromium, copper, lead, mercury, and zinc in sediments collected in Bayou d'Inde, southwestern Louisiana. AB - The concentrations of the metals chromium (Cr), copper (Cu), lead (Pb), mercury (Hg), and zinc (Zn) were determined in sediment cores taken from Bayou d'Inde in southwestern Louisiana. Previous studies had reported elevated concentrations of these trace metals in sediments along this waterway. The current study, conducted more than a decade later, was designed to determine if these metals had migrated from the areas of highest concentrations. Concentrations as high as 400 mg/kg for Cr, 1100 mg/kg for Cu, 10 mg/kg for Hg, and 600 mg/kg for Pb and Zn were found. Areas of high concentrations of metals were isolated within a distance of about 1 km from the most highly contaminated areas in the bayou. Low concentrations of metals were found at the mouth of the bayou. Estimated sedimentation rates of 0.67-1.2 cm/yr were based on the burial of the sediments originally studied. The concentrations of the metals studied remained high and were also highly localized, both spatially and temporally. PMID- 11878607 TI - Wind tunnel simulation of air pollution dispersion in a street canyon. AB - Physical simulation was used to study pollution dispersion in a street canyon. The street canyon model was designed to study the effect of measuring flow and concentration fields. A method of C02-laser photoacoustic spectrometry was applied for detection of trace concentration of gas pollution. The advantage of this method is its high sensitivity and broad dynamic range, permitting monitoring of concentrations from trace to saturation values. Application of this method enabled us to propose a simple model based on line permeation pollutant source, developed on the principle of concentration standards, to ensure high precision and homogeneity of the concentration flow. Spatial measurement of the concentration distribution inside the street canyon was performed on the model with reference velocity of 1.5 m/s. PMID- 11878608 TI - Committee on Pesticides and Disinfectant Formulations. Pesticide formulations: CIPAC studies. Collaborative International Pesticide Analytical Council. PMID- 11878609 TI - Determination of mercury in sewage sludges by slurry sampling electrothermal atomic absorption spectrometry. AB - An electrothermal atomic absorption spectrometric procedure was developed for the determination of mercury in sewage sludge, in which the samples were suspended in a solution containing hydrofluoric and nitric acids. Silver nitrate (4%, m/v) and potassium permanganate (3%, m/v) were incorporated as matrix modifiers, and aliquots were directly introduced into the graphite furnace. A fast-heating program with no conventional pyrolysis step was used. The detection limit for mercury in a 50 mg/mL suspension was 0.1 microg/g. Calibration was performed by using aqueous standards. An analysis of certified reference materials confirmed the reliability of the procedure. PMID- 11878610 TI - Committee on Drugs and Related Topics. Drug residues in animal tissues. PMID- 11878611 TI - Committee on Residues and Related Topics. Metals and other elements. PMID- 11878612 TI - Committee on Residues and Related Topics. Multiclass multiresidue methods for organic compounds. PMID- 11878613 TI - Committee on Residues and Related Topics. Single class multiresidue methods for organic compounds. PMID- 11878614 TI - Committee on Microbiology and Extraneous Materials. Food microbiology--non-dairy. PMID- 11878615 TI - Committee on Feeds, Fertilizers, and Related Agricultural Topics. Feeds. PMID- 11878616 TI - Determination of arsenic in homeopathic drugs by bromide volatilization inductively coupled plasma-atomic emission spectrometry. AB - Arsenic in homeopathic drugs was determined by coupling a volatile generation with inductively coupled plasma-atomic emission spectrometry. The method is based on the chemical vaporization of arsenic(III) with bromide ions in sulfuric acid media using a batch procedure and subsequent introduction of the gaseous analyte into the plasma torch. The main and interactive effects of the experimental variables affecting this method were evaluated by a 2-level full factorial design. In optimized conditions by Simplex, the method shows an absolute detection limit (3 s) of 0.28 microg for the injection of 230 microL sample. The precision (% relative standard deviation) of the determination was 4.2% at a level of 50 microg/mL As(III) (n = 5). The interference effect of various ions on the arsenic signal was evaluated. PMID- 11878618 TI - Determination of organochlorinated compounds in marine organisms by microwave assisted extraction with molecular organized systems and liquid chromatography with fluorescence detection. AB - Microwave-assisted extraction methodology is used to extract different compounds from various kinds of marine solid samples, such as soils, sediments, and organisms. A new analytical method was developed to extract polychlorinated biphenyls and polychlorinated dibenzofurans by using a conventional microwave system and the nonionic surfactant, polyoxyethylene 10 lauryl ether as the extractant as a prior step to liquid chromatography analysis coupled with fluorescence detection. The method was applied to the extraction and determination of these analytes in samples of blue mussels (Mytilus galloprovincialis), cockles (Cerastoderma edule), and clams (Dosinia exoleta). Compared with the traditional Soxhlet extraction, results of the proposed method showed acceptable recovery percentages for the organochlorinated compounds under study and standard deviation values <10%. PMID- 11878617 TI - Size exclusion chromatographic cleanup for GC/MS determination of organophosphorus pesticide residues in household and vehicle dust. AB - Size exclusion chromatography (SEC) was used as a cleanup method for the analysis of organophosphorus pesticides in household and vehicle dusts. The pesticides investigated were diazinon, methyl parathion, chlorpyrifos, malathion, phosmet, and azinphosmethyl. These compounds are of interest due to their use in agricultural tree fruit production and/or urban pest control. Pesticides were determined via gas chromatography/mass spectrometry with selected-ion monitoring and cool on-column injection. The lower limit of method validation was 0.20 microg/g. Method limits of detection in dust ranged from 0.012-0.055 microg/g. Dust samples were collected with vacuums from the homes and vehicles of people living and working in a rural agricultural region in the central part of Washington State. The analytes were extracted from the dust by sonication in acetone. The extracts were solvent-exchanged to cyclohexane, frozen, thawed, and centrifuged prior to SEC injection. Following SEC, the eluent was split into 2 fractions, concentrated, and injected on-column into the gas chromatograph. This method represents the first complete publication describing the SEC cleanup of organophosphorus pesticides in dusts. Recoveries of pesticides in dusts ranged from 63.5-110.8 +/- 4.9-19.6% over a fortification range of 0.20-10.00 microg/g. This optimized, automated, and reproducible SEC method does not require further treatment or cleanup for trace determination of these organophosphorus pesticides. PMID- 11878619 TI - Gas chromatographic and mass spectrometric analysis of nitrilotriacetic acid in environmental aqueous samples. AB - This study describes a fast and accurate method for the sample preparation, identification, and quantitation of nitrilotriacetic (NTA) acid in environmental aqueous samples at a concentration of ppb level. The method is sensitive, specific, and free from the interferences of fatty and amino acids. The tri-n propyl- and tri-n-butyl-NTA acid esters were prepared by the reaction of n-propyl HCl and n-butyl-HCl solutions and NTA acid, respectively. The derivatives were analyzed by a gas chromatograph equipped with a mass spectrometric detector. The method detection limit, 0.006 mg/L of each NTA ester, was determined and validated by an analysis of a fortified water sample. The overall recoveries were 103-115%, n = 8. The method was applied to a real sample and a 0.90 mg/L concentration of NTA acid was found. Mass fragmentation patterns of the derivatives are also reported. PMID- 11878620 TI - Dry rehydratable film method for rapid enumeration of coliforms in foods (3M Petrifilm Rapid Coliform Count plate): collaborative study. AB - A rehydratable dry-film plating method for coliforms in foods, the 3M Petrifilm Rapid Coliform Count plate method, was compared with the U.S. Food and Drug Administration's Bacteriological Analytical Manual method for nondairy foods and the American Public Health Association's Standard Methods for the Examination of Dairy Products (SMEDP) method for dairy foods. Six food types, vanilla ice cream, cheddar cheese, fresh refrigerated uncooked pasta, wheat flour, prepared frozen macaroni and cheese, and frozen hash browns, were analyzed for coliforms by 11 collaborating laboratories. For each food product tested, the collaborators received 8 blind samples consisting of a control sample and 3 levels of inoculated sample, each in duplicate. The mean log counts for the methods were comparable. The repeatability and reproducibility variances of the Petrifilm Rapid Coliform Count method at 14 and 24 h were not significantly different from those of the standard methods. PMID- 11878621 TI - McCleary wins 2002 Wiley Award. PMID- 11878622 TI - Residues of oxytetracycline and its 4'-epimer in edible tissues from turkeys. AB - Residues of oxytetracycline (OTC) in edible tissues (muscle, liver, and kidney) of 18 turkeys were determined after continuous administration of the drug for 3 days in drinking water at the maximum recommended concentration of 400 mg/L. The European Union (EU) maximum residue limits (MRLs) set for OTC are 100 microg/kg in muscle tissues, 300 microg/kg in liver, and 600 microg/kg in kidney, as the sum of the parent compound and its derivative 4'-epi-oxytetracycline (4-epi-OTC). Cleanup of tissue samples was performed by metal chelate affinity chromatography (MCAC), but the original technique was miniaturized by the adoption of a mini solid-phase extraction column, allowing reduction of solvents, time, and hazardous waste. OTC and its 4'-epimer were quantitated by an isocratic liquid chromatography elution with UV detection. After 1 day of withdrawal, OTC plus 4 epi-OTC residues were greater than MRL values in muscle and liver; 3 days after the end of treatment, all tissue residues were far lower than the MRL values. At the first day after the end of treatment, 4-epi-OTC was detected at very low concentrations only in muscle, in liver after 1 and 3 days of withdrawal, and in kidney at all sampling times. The withdrawal time was calculated according to EU recommendations and was set at 5 days. PMID- 11878623 TI - Determination of capsaicinoids in salsa by liquid chromatography and enzyme immunoassay. AB - Two simple and rapid methods were developed to monitor pungency of salsa in production. Capsaicin (C) and dihydrocapsaicin (DHC) were quantitated in 17 commercially available tomato-based salsas by enzyme immunoassay (EIA) and liquid chromatography (LC) with fluorescent detection. Samples were extracted with methanol and the extracts were subjected to solid-phase extraction (SPE) using polystyrene-divinylbenzene columns. Analysis of SPE eluates showed good correlation (r2 = 0.953) between LC and EIA, with a slightly high bias for EIA. Salsa fortified with C and DHC from 0.118 to 103.2 microg/g resulted in recoveries of 90-112% (C) and 76-97% (DHC). Limits of detection by LC were 0.1 microg/g for each capsaicinoid and 0.1 microg/g by EIA for total capsaicinoids. The LC on-column response was linear from 0.2 to 100 ng for both C and DHC, whereas the working range for EIA was 0.1-2.0 ppm. Pungency varied between different salsa brands labeled mild, medium, and hot. PMID- 11878624 TI - Gas chromatographic analysis of infant formulas for total fatty acids, including trans fatty acids. AB - Twelve powdered and 13 liquid infant formulas were analyzed by using an extension of AOAC Official Method 996.01 for fat analysis in cereal products. Samples were hydrolyzed with 8 N HCl and extracted with ethyl and petroleum ethers. Fatty acid methyl esters were prepared by refluxing the mixed ether extracts with methanolic sodium hydroxide in the presence of 14% boron trifluoride in methanol. The extracts were analyzed by gas chromatography. In powdered formulas, saturated fatty acid (SFA) content (mean +/- SD; n = 12) was 41.05 +/- 3.94%, monounsaturated fatty acid (MUFA) content was 36.97 +/- 3.38%, polyunsaturated fatty acid (PUFA) content was 20.07 +/- 3.08%, and total trans fatty acid content was 1.30 +/- 1.27%. In liquid formulas, SFA content (mean +/- SD; n = 13) was 42.29 +/- 2.98%, MUFA content was 36.05 +/- 2.47%, PUFA content was 20.65 +/- 2.40%, and total trans fatty acid content was 0.88 +/- 0.54%. Total fat content in powdered formulas ranged from 4.4 to 5.5 g/100 kcal and linoleic acid content ranged from 868 to 1166 mg/100 kcal. In liquid formulas, total fat content ranged from 4.1 to 5.1 g/100 kcal and linoleic acid content ranged from 820 to 1100 mg/100 kcal. There were no significant differences between powdered and liquid infant formulas in concentrations of total fat, SFA, MUFA, PUFA, or trans fatty acids. PMID- 11878625 TI - Determination of low-level glucose and fructose in raw and refined crystalline sugar by high-performance anion-exchange chromatography: collaborative study. AB - A method was developed and a collaborative study was performed under the auspices of the International Commission of Uniform Methods for Sugar Analysis. The collaborators used high-performance anion-exchange chromatography (HPAEC) to determine trace amounts of glucose and fructose in 3 raw and 3 refined sugar samples provided as blind duplicates. Fourteen laboratories participated in the study. Although difficulties were experienced by a few analysts, 10 laboratories reported positive results. Average repeatabilities and reproducibilities for glucose and fructose in raw sugar were slightly above 5 and 10%, respectively, and the average Horwitz ratios were well under 2. Average repeatabilities and reproducibilities for glucose and fructose in refined sugar were 10 and 22%, respectively; although the Horwitz ratios were >2, they were marginal (2.8). The HPAEC results agreed with results obtained by gas chromatography in an independent laboratory. Because the method was collaboratively studied according to the protocol of the International Union of Pure and Applied Chemistry for collaborative studies, and the results meet its criteria, it is recommended that the method be adopted Official First Action by AOAC INTERNATIONAL. PMID- 11878626 TI - Atmospheric dust loads and their elemental composition at a background site in India. AB - Air particulate samples collected during 1995-96 at a background site situated on the east coast of Thar Desert in Rajsthan State of India were analysed for atmospheric dust loads (Suspended Particulate Matter) and elemental composition. The values of SPM ranged from 9 microg M(-3) to 97 microg M(-3) with an average of 43 microg M(-3) except a few episodic values, which were 3 to 5 times higher than the average during summer months. The results for elemental composition of the particulate samples showed that the concentrations of anthropogenic toxic trace elements viz. Br, Cr, Pb, Sb, Se and Zn are lower by a factor of 2 to 10 as compared to urban areas. The high enrichment factors for anthropogenic elements viz. Br, Pb, Sb and Zn show an input from coal/wood fuel burning and vehicular pollution at the sampling site. The depletion of Si in SPM samples shows long distance transport of dust to the sampling site. PMID- 11878627 TI - Forest area in Costa Rica: a comparative study of tropical forest cover estimates over time. AB - Forest area figures. at a given point in time and for a given region of interest, differ considerably, affecting the calculation of deforestation rates and thus confuse the political and scientific discussion on the state and change of the resource forest. This article discusses the variation of published forest cover figures, using Costa Rica as an example. A list of published figures on the forest cover of Costa Rica from 1940 onwards is analyzed. Reasons for the differences are hypothesized and discussed. These differences are mainly in the definition of forest and forest classes included, in the type of the studies conducted (mapping and/or sampling), in the precision of the estimates, and in the information sources used. It is concluded that part of the problem is inherent in the nature of the resource 'forest'. Quality and completeness of the presentation of the forest cover estimates are a clue to their correct understanding and interpretation. The latter point being especially relevant, as forest cover data have both a technical-scientific and a political meaning and are used as relevant arguments in many discussions. In the example of Costa Rica, a general downward trend is observed up to about 1985/1990, whereas after that forest area figures are on the average at a markedly higher level. Some hypotheses for this change in the trend are discussed. PMID- 11878628 TI - Necessity of toxicity assessment in Turkish industrial discharges (examples from metal and textile industry effluents). AB - Toxicity of some organic and inorganic chemicals to microorganisms is an important consideration in assessing their environmental impact against their economic benefits. Microorganisms play an important role in several environmental processes, both natural and engineered. Some organic and inorganics at toxic levels have been detected in industrial discharges resulting in plant upsets and discharge permit violations. In addition to this, even though in some cases the effluent wastewater does not exceed the discharge limits, the results of toxicity tests show potential toxicity. Toxicity knowledge of effluents can benefit treatment plant operators in optimising plant operation, setting pre-treatment standards, and protecting receiving water quality and in establishing sewer discharge permits to safeguard the plant. In the Turkish regulations only toxicity dilution factor (TDF) with fish is part of the toxicity monitoring program of permissible wastewater discharge. In various countries, laboratory studies involving the use of different organisms and protocol for toxicity assessment was conducted involving a number of discharges. In this study, it was aimed to investigate the acute toxicity of textile and metal industry wastewaters by traditional and enrichment toxicity tests and emphasize the importance of toxicity tests in wastewater discharge regulations. The enrichment toxicity tests are novel applications and give an idea whether there is potential toxicity or growth limiting and stimulation conditions. Different organisms were used such as bacteria (Floc and Coliform bacteria) algae (Chlorella sp.). fish (Lepistes sp.) and protozoan (Vorticella sp.) to represent four tropic levels. The textile industry results showed acute toxicity for at least one organism in 8 out of 23 effluent samples. Acute toxicity for at least two organisms in 7 out of 23 effluent sampling was observed for the metal industry. The toxicity test results were assessed with chemical analyses such as COD, BOD, color and heavy metals. It was observed that the toxicity of the effluents could not be explained by using physicochemical analyses in 5 cases for metal and 4 cases for the textile industries. The results clearly showed that the use of bioassay tests produce additional information about the toxicity potential of industrial discharges and effluents. PMID- 11878629 TI - Relationships of human disturbance, bird communities, and plant communities along the land-water interface of a large reservoir. AB - We examined the relationships of human activity, bird communities, and plant communities along the land-water interface of Lake Texoma, a large human-made reservoir on the Texas-Oklahoma border. Measurements of human activity, plant surveys, and bird surveys were performed at 40 paired transects, one with human disturbance, the other undisturbed. Both principal components and correspondence analyses of bird-survey data separated disturbed sites from paired undisturbed sites, and typical disturbance-tolerant species from those less tolerant of human activity. Compared to undisturbed sites, disturbed sites tended to have more individual birds per survey, pavement, and mowed lawns, and less canopy, vegetation volume, and vegetation vertical diversity. A principal components analysis of quantitative disturbance measurements revealed that most bird and plant measures were highly correlated with the first disturbance component. However, the correlation between birds and human activity was much stronger than that between birds and plants, or between plants and disturbance. Our data suggest that bird-species composition is regulated more by human activity than by plant-community composition. Also, in this system, bird communities are a better choice than plant communities to index the effect of human disturbance. To maintain regional diversity of both birds and plants, undisturbed areas should be maintained around reservoirs. PMID- 11878630 TI - Prediction of leachate level in Kimpo metropolitan landfill site by total water balance. AB - Kimpo metropolitan landfill has received various kinds of wastes since January 1992. The leachate level was measured to be 10.3 m in May 1995 and the level increased to 12.2 m in August 1996. Therefore, to prove the reason for the increasing leachate level, we calibrated hydraulic conductivity of each waste and intermediate layer using the HELP (Hydrologic Evaluation of Landfill Performance) model. The leachate generation data measured from February 1993 to October 1995 was used in the model calibration. As a result of a model calibration, we obtained an average infiltration ratio and used this in analysis of the total water balance to predict elevation of leachate level. Main causes of the elevation of the leachate level were the high water content of the waste and the degradation of the leachate-drainage system caused by the subsidence of a natural barrier layer. PMID- 11878631 TI - Heavy metal accumulation in lichens from the Hetauda industrial area Narayani zone Makwanpur District, Nepal. AB - Abstract. Lichen samples collected in and around Hetauda Industrial area, (HIA) Narayani zone, Makwanpur district, Nepal, were analyzed for Cr, Ni, Pb, Zn, Ca, Mn, Fe, Si, and Al. The samples from the location inside the industrial area have higher levels of metal than the outside areas. Pyxine meissnerina growing inside the industrial area accumulated higher levels of all the metals analyzed. PMID- 11878632 TI - Cadmium contamination in wild birds as an indicator of environmental pollution. AB - Cadmium (Cd) is an environmental pollutant that has serious toxicity in humans and animals and causes Itai-Itai disease. However, there is little available information on its contamination in wildlife as an indicator of environmental pollution. The Cd contents in the kidney and liver of 85 wild birds from 9 different prefectures in Japan were investigated. The ranges of the Cd contents in the kidney and liver in all birds were ND-174.4 and ND-21.2 microg g(-1) dry wt., respectively. The mean Cd contents were higher in the oil-contaminated birds than those in the non-contaminated ones. Furthermore, a strong correlation was obtained only between the Cd contents in the kidney and those in the liver of the oil-contaminated seabirds and not in the other non-contaminated ones. These results suggest that wild birds reflect the level of environmental contamination which should be monitored. PMID- 11878633 TI - Spatial patterns of soil moisture as affected by shrubs, in different climatic conditions. AB - Abstract. At three study sites, representing Mediterranean, semi-arid and mildly arid climatic conditions, the effect of shrubs on the spatial patterns of soil moisture was studied. At each site soil moisture was measured, on hillslopes, at the vicinity of 8 shrubs. For each shrub the measurements have been taken at 3 microenvironments, i.e. under the shrub (US), at the margins of shrub (MS) and between shrubs (BS). At the microenvironments US and MS the measurements were taken at 3 directions: upslope, downslope and sideslope of the shrubs. At all sampling points soil samples were taken from 3 depths: 0-2, 2-5 and 5-10 cm. In addition, rock fragments cover percentage near the shrubs was determined. A soil moisture pattern was found, around each shrub, which is composed of a radial gradient and a downslope gradient. The radial gradient is expressed by soil moisture decreasing from the US microenvironment, in all directions, through the MS towards the BS microenvironment. The US microenvironment has a 'spatial advantage' of higher soil moisture content due to (1) relatively higher infiltration rate, (2) capture overland flow from the BS area upslope that shrub and (3) low evaporation rate because of the shading effect. The downslope gradient is expressed by decreasing soil moisture from the upslope direction of each shrub (MS and US microenvironments) towards the downslope direction of that shrub (MS and US microenvironments, respectively). This gradient is controlled by the relatively high content of rock fragments near the shrubs at their upslope direction. Such rock fragments spatial distribution is attributed to (1) the detachment and transport of rock fragments by sheep and goats trampling and (2) the effect of shrub on the continuity of overland flow and sediment transport. The effect of rock fragments is similar to that of shrubs regarding increasing infiltration and decreasing evaporation rate. The relatively high soil moisture at the upslope direction of each shrub enhances annuals growth producing a positive feedback loop: soil moisture--annuals growth--trampling. This sequence maintains the typical rock fragments spatial organization and contributes to the sustainability of the grazing system. At all the study sites at the US microenvironment there is a trend of decreasing soil moisture with increasing soil depth. At microenvironments MS and BS soil moisture increases with soil depth. The results are of great relevance for rehabilitation strategies as they suggest that in order to combat desertification in degraded semi-arid and mildly arid areas, where the main land use is grazing, both shrubs and rock fragment should be kept at their present spatial distribution. PMID- 11878634 TI - Trihalomethane formation potential and concentration changes during water treatment at Mumbai (India). AB - The treated water at the outlet of treatment plants and representative service reservoirs of Mumbai city have been evaluated for trihalomethane formation potential in 1995-1996. Chloroform, dichlorobromomethane, chlorodibromomethane and bromoform have been monitored during monsoon, winter and summer. The levels of chloroform are found above the regulated WHO guideline value of 200 microg L( 1) in final water during postmonsoon at Ghatkopar (226 microg L(-1)), Malbar (210.3 microg L(-1)) and Tulsi (231.26 microg L(-1)). PMID- 11878635 TI - Seasonal variation in bacterial flora of the wastewater and soil in the vicinity of industrial area. AB - Environmental pollution is one of the major problems being faced today due to the advent of extensive industrialization. Heavy metals are the most important contaminants in wastewater that are present in abundance and are toxic. Heavy metal contents of sewage in the industrial estate of Aligarh (U.P.) have been determined by atomic absorption spectrophotometry. The analysis of samples collected from six different locations revealed significantly high levels of Fe, Zn, Cu, Cr and Ni. Certain bacteriological (Total bacterial count, Total coliform. Fecal coliform, and Fecal Streptococci) parameters of domestic and industrial sewage as well as soils were monitored from March 1990 to January 1993. Total bacterial count, total coliform, fecal coliform and fecal streptococci were found to be lowest in all the samples of industrial wastewater compared to those in domestic sewage and soil systems. The soil however, contained highest total culturable bacterial population. In view of the common practice of the application of sewage to agricultural land in the neighbouring area, the discharge of industrial wastewater without proper treatment into public sewers should be strictly prohibited. PMID- 11878636 TI - Soil-water partitioning and desorption hysteresis of volatile organic compounds from a Louisiana Superfund site soil. AB - The adsorption and desorption of three volatile organic compounds (1,2 dichloroethane, 1,1,2- trichloroethane and 1,1,2,2-tetrachloroethane) from a previously uncontaminated clayey soil sample from a Superfund site in North Baton Rouge, Louisiana was studied. In the linear range of the adsorption isotherm, the partition constants were not affected by the presence of the co-solutes. The adsorption isotherms over a wide concentration range on the soil followed the nonlinear Freundlich isotherm. The desorption of the compounds showed significant hysteresis at all concentrations studied. Approximately 20 to 70% of the adsorbed mass of organic compounds resisted the desorption even after five months of successive desorption steps. The desorption of four compounds (1,2 dichloroethane, 1,1,2-trichloroethane, 1,4-dichlorobenzene and hexachlorobutadiene) from a contaminated soil sample from the same site was also studied. The aqueous concentration declined as the successive desorption steps progressed. For hexachlorobutediene the desorption can be visualized as occurring in two stages. The first stage involved a 'loosely bound' or 'reversible' fraction and the second stage involved a 'tightly bound' or 'resistant' fraction. PMID- 11878637 TI - External contribution to urban air pollution. AB - Elevated particulate matter concentrations in urban locations have normally been associated with local traffic emissions. Recently it has been suggested that such episodes are influenced to a high degree by PM10 sources external to urban areas. To further corroborate this hypothesis, linear regression was sought between PM10 concentrations measured at eight urban sites in the U.K., with particulate sulphate concentration measured at two rural sites, for the years 1993-1997. Analysis of the slopes, intercepts and correlation coefficients indicate a possible relationship between urban PM10 and rural sulphate concentrations. The influences of wind direction and of the distance of the urban from the rural sites on the values of the three statistical parameters are also explored. The value of linear regression as an analysis tool in such cases is discussed and it is shown that an analysis of the sign of the rate of change of the urban PM10 and rural sulphate concentrations provides a more realistic method of correlation. The results indicate a major influence on urban PM10 concentrations from the eastern side of the United Kingdom. Linear correlation was also sought using PM10 data from nine urban sites in London and nearby rural Rochester. Analysis of the magnitude of the gradients and intercepts together with episode correlation analysis between the two sites showed the effect of transported PM10 on the local London concentrations. This article also presents methods to estimate the influence of rural and urban PM10 sources on urban PM10 concentrations and to obtain a rough estimate of the transboundary contribution to urban air pollution from the PM10 concentration data of the urban site. PMID- 11878638 TI - Aerosol scavenging: model application and sensitivity analysis in the Indian context. AB - Sulfate aerosols have been found to be the major contributors to precipitation acidity. Thus, in view of the long-term ecological repercussions they have on aquatic ecosystems and their acidity-potential, the present analysis focuses on a case study application of the layer-averaged aerosol-scavenging model (Okita et al., 1996) for predicting values of the wet scavenging coefficient and sulfate concentrations in precipitation samples on the basis of the information available for some selected Indian cities. Through sensitivity analysis (Pandey et al., 1997) the scavenging coefficient has been found to be very strongly dependent on precipitation intensity. Comparison of model predictions has been done with the measured values for Delhi, Mumbai, Calcutta and Chennai in India. PMID- 11878639 TI - Assessment of climate change effects on Canada's National Park system. AB - To estimate the magnitude of climate change anticipated for Canada's 38 National Parks (NPs) and Park Reserves, seasonal temperature and precipitation scenarios were constructed for 2050 and 2090 using the Canadian Centre for Climate Modelling and Analysis (CCCma) coupled model (CGCM1). For each park, we assessed impacts on physical systems, species, ecosystems and people. Important, widespread changes relate to marine and freshwater hydrology, glacial balance, waning permafrost, increased natural disturbance, shorter ice season, northern and upward altitudinal species and biome shifts, and changed visitation patterns. Other changes are regional (e.g., combined East coast subsidence and sea level rise increase coastal erosion and deposition, whereas, on the Pacific coast, tectonic uplift negates sea level rise). Further predictions concern individual parks (e.g., Unique fens of Bruce Peninsular NP will migrate lakewards with lowered water levels, but structural regulation of Lake Huron for navigation and power generation would destroy the fens). Knowledge gaps are the most important findings. For example: we could not form conclusions about glacial mass balance, or its effects on rivers and fjords. Likewise, for the East Coast Labrador Current we could neither estimate temperature and salinity effects of extra iceberg formation, nor the further effects on marine food chains, and breeding park seabirds. We recommend 1) Research on specific large knowledge gaps; 2) Climate change information exchange with protected area agencies in other northern countries; and 3) incorporating climate uncertainty into park plans and management. We discuss options for a new park management philosophy in the face of massive change and uncertainty. PMID- 11878640 TI - Effects of scale and logging on landscape structure in a forest mosaic. AB - Landscape structure in a forest mosaic changes with spatial scale (i.e. spatial extent) and thresholds may occur where structure changes markedly. Forest management alters landscape structure and may affect the intensity and location of thresholds. Our purpose was to examine landscape structure at different scales to determine thresholds where landscape structure changes markedly in managed forest mosaics of the Appalachian Mountains in the eastern United States. We also investigated how logging influences landscape structure and whether these management activities change threshold values. Using threshold and autocorrelation analyses, we found that thresholds in landscape indices exist at 400, 500, and 800 m intervals from the outer edge of management units in our study region. For landscape indices that consider all landcover categories, such as dominance and contagion, landscape structure and thresholds did not change after logging occurred. Measurements for these overall landscape indices were strongly influenced by midsuccessional deciduous forest, the most common landcover category in the landscape. When restricting analyses for mean patch size and percent cover to individual forest types, thresholds for early successional forests changed after logging. However, logging changed the landscape structure at small spatial scale, but did not alter the structure of the entire forest mosaic. Previous forest management may already have increased the heterogeneity of the landscape beyond the point where additional small cuts alter the overall structure of the forest. Because measurements for landscape indices yield very different results at different spatial scales, it is important first to identify thresholds in order to determine the appropriate scales for landscape ecological studies. We found that threshold and autocorrelation analyses were simple but powerful tools for the detection of appropriate scales in the managed forest mosaic under study. PMID- 11878641 TI - Using mussel isotope ratios to assess anthropogenic nitrogen inputs to freshwater ecosystems. AB - Stable nitrogen isotope ratios (delta15N) of freshwater mussels from a series of lakes and ponds were related to watershed land use characteristics to assess their utility in determining the source of nitrogen inputs to inland water bodies. Nitrogen isotope ratios measured in freshwater mussels from 19 lakes and ponds in Rhode Island, U.S.A., ranged from 4.9-12.6 per thousand and were found to significantly correlate with the fraction of residential development in 100 and 200 m buffer zones around the ponds. Mussel delta15N values in 12 of the 19 ponds also showed significant correlation with average dissolved nitrate concentrations, which ranged from 23-327 microg L(-1). These observations, in light of previous studies which link elevated delta15N values of nitrogen derived from septic wastewater with those seen in biota, suggest that mussel isotope ratios may reflect nitrogen source in freshwater ecosystems. We followed an iterative approach using multiple regression analysis to assess the relationship between mussel delta15N and the land use categories fraction residential development, fraction feedlot agriculture, fraction row-crop agriculture, and fraction natural vegetation in 100 and 200 m buffer zones and pond watersheds. From this we developed a simple regression model to predict mussel delta15N from the fraction of residential development in the 200 m buffer zone around the pond. Subsequent testing with data from 16 additional sites in the same ecoregion led us to refine the model by incorporating the fraction of natural vegetation. The overall average absolute difference between measured and predicted delta15N values using the two-parameter model was 1.6 per thousand. Potential sources of error in the model include differences in the scale and categorization of land use data used to generate and test the model, differences in physical characteristics, such as retention time and range of residential development, and exclusion of sources of enriched nitrogen such as runoff from feedlot operations or increased nitrogen loading from inefficient or failed septic systems. PMID- 11878642 TI - Trace element contamination in benthic macroinvertebrates from a small stream near a uranium mill tailings site. AB - Direct measurement of the accumulation of non-radioactive trace elements in aquatic biota near uranium mining or processing sites has been relatively rare, with greater focus on the radiological activity in the adjacent soils and groundwater. To evaluate the potential ecological concern associated with trace elements at a former uranium mill site in southeastern Utah, benthic macroinvertebrates were collected and analyzed for 17 trace elements from multiple locations within a small on-site stream, Montezuma Creek, and a nearby reference stream. Key questions of this study relate to the spatial and temporal extent of contamination in aquatic biota, the potential ecological risks associated with that contamination, and the usefulness of benthic macroinvertebrates as a monitoring tool at this site. Composite samples of similar macroinvertebrate taxa and functional feeding groups were collected from each site over a two year period that was representative of normal and dry-year conditions. In both years, mean concentrations of arsenic, molybdenum, selenium, and vanadium were significantly higher (a factor of 2-4 times: P < 0.05) in macroinvertebrates collected from one or both of the two Montezuma Creek sites immediately downstream of the mill tailing site in comparison to concentrations from reference locations. Mean uranium concentrations in invertebrates immediately downstream of the mill site were more than 10 times higher than at reference sites. The site-to-site pattern of contamination in Montezuma Creek invertebrates was similar in 1995 and 1996, with mill-related trace elements showing a downstream decreasing trend. However, nine of seventeen contaminant concentrations were higher in the second year of the study, possibly due to a higher influx of deep groundwater during the drier second year of the study. A preliminary assessment of ecological risks, based on the benthic macroinvertebrate bioaccumulation data, suggests that aquatic and terrestrial population risks are low. Benthic macroinvertebrates appeared to be sensitive integrators of trace element inputs to the aquatic environment from a former uranium mill tailing site, and provided useful spatial and temporal patterns of contamination not easily obtained using conventional surface water or groundwater measures. PMID- 11878643 TI - Uranium, thorium, and potassium in soils along the shore of Lake Issyk-Kyol in the Kyrghyz Republic. AB - The Kyrghyz Republic, located in the southeastern region of the former Soviet Union, maintains a population of more than one-half-million persons and is heavily dependent on Lake Issyk-Kyol, both to draw tourists to the area and for its utilization by some as a food and recreation source. Historical surveys, conducted primarily for geological exploration, have indicated that localized areas of shoreline on Lake Issyk-Kyol have relative radiation levels in excess of ambient background by as much as a factor often. Uranium mining operations in the mountains bordering the lake to the south may have resulted in the contamination of a number of areas on the lake's southern shore. Concentrations of naturally occurring uranium, thorium, and potassium are present in these soils in elevated quantities. This paper presents the results of an investigation of soil concentrations along the shoreline of Lake Issyk-Kyol relative to previously discovered areas of high exposure rate. PMID- 11878644 TI - Determination of fish community composition in the untempered regions of a thermal effluent canal--the efficacy of a fixed underwater videography system. AB - High flows and deep waters associated with thermal discharge canals invoke safety concerns and preclude the use of conventional fish sampling methodologies. Despite these challenges, it is critical that assessments of thermal effects from power generation do not rely solely on data collected in the plume, but also focus on fish living in the canal, particularly in regions above the zone of tempering influence. We deployed a fixed underwater videography apparatus to monitor the community composition and abundance patterns of fish in the Nanticoke thermal generating discharge canal on the north shore of Lake Erie from February 1, 1999 until July 31, 1999. We also compared the number of species observed using video to angling surveys, visual observations through the air/water interface and two modified netting procedures. Our results indicate that videography permitted the detection of the most species and the highest number of individuals. Variable visibility was the largest limitation of this approach. The addition of infrared lighting for low light conditions and a series of cameras positioned at several depths would be a cost effective, safe, and efficient method of assessing community structure and behaviour of fish in thermal discharge canals. PMID- 11878645 TI - Monitoring of aquatic macrophytes for detection of long-term change in river systems. AB - This paper presents details of the methodology developed by the United Kingdom's Environmental Change Network for the long-term monitoring of macrophytes in rivers and streams. The methodology is based on techniques first proposed by the Standing Committee of Analysts (1987) and later adapted by the National Rivers Authority (NRA) and Environment Agency, but differs in splitting the surveyed 100 m stretch of water into sections to provide an objective measure of the frequency of occurrence of individual species in place of the more subjective estimation of cover. A pilot study of the ECN methodology took place at five sites in 1997. The results of this study, including a few practical difficulties in the application of the methodology, are presented and discussed. For all but one of the sites strong associations were found between the number of species observed and the physical characteristics of the watercourse. The most important characteristics were degree of shading, substrate type, depth and clarity. The frequency of occurrence of individual species within sections of the watercourse was found to be strongly related to the log of the overall estimates of cover. Because the use of sections, rather than a single overall cover estimate, enables variation in the pattern of vegetation over surveyed stretches to be detected and related to watercourse characteristics, the precision with which change can be detected is increased, and the possibility of determining the causes of change is thereby enhanced. Moreover the use of sections allows within-site variation to be calculated and hence the accuracy of estimated changes to be quantified. In general implementation of the ECN methodology was not found to be particularly onerous or difficult. As a result of the pilot study some changes in the ECN methodology have been made, primarily to reduce the workload so that sites can be surveyed comfortably in a single day. PMID- 11878646 TI - Potential effects of metals in reacidified limed water bodies in Norway and Sweden. AB - The goal of this work was to assess risk of chemical and biological effects of metals in reacidified, limed water bodies in Norway and Sweden. The risk assessment is based on a literature review and evaluations of water chemical data from the 1995 Nordic Lake Survey. Compared to the pre-liming period, it us unlikely that enhanced remobilization of inorganic aluminium (Al) or other toxic metals (metal bomb hypothesis) from the catchment, the lake sediment and/or the streambed will occur when limed waters reacidify. Rather, the concentrations in surface waters are expected to be lower than before liming started, because of reduced atmospheric inputs of both strong acids and metals as Cd, Hg, Pb, and Zn during the last 10-20 yr. The concentrations in lakes relative to the biological effect levels, as well as the chemical properties of the different metals suggest that the potential biological risks associated to reacidification of limed lakes decrease in the order Al >> Cd > Pb. The risks associated with Cr, Cu, Fe, Mn, Ni and Zn are very low and do not have to be considered except in waters with known concentrations larger than the lowest biological risk level. Such waters are very rare (<2%). Aluminium is the metal that should be used to set the limit for judging the risk of biological damage due to reacidification of limed surface waters. PMID- 11878647 TI - A simplified liquid-solid extraction technique for the analyses of pesticide residues in soil samples. AB - A simplified liquid-solid extraction technique was studied for the analyses of pesticide residues in soil samples. It is a simple, one step sample preparation method based on a relative quantification concept. Linear response curves were obtained for all of the target compounds regardless of their surface adsorption. This observation was explained and modeled to obey Langmuir adsorption Equation. The adsorption of analytes onto the sample surfaces will sacrifice the detection sensitivity. The strategies to reduce the surface adsorption, such as, molecular replacement and selection of solvents, were discussed. The relative quantification and the wide varieties of available solvents would enable the technique to be a useful method for the monitoring and analyses of pesticide residues in soils. PMID- 11878648 TI - Inorganic ion levels of soils and streams in some areas of Ogoniland, Nigeria as affected by crude oil spillage. AB - The inorganic ion concentrations of soils and streams in the three locations (Yorla, Zaakpon and Goi) in Ogoniland, Nigeria affected by crude oil spillage were investigated. In general, inorganic ion concentrations of polluted soils and streams varied significantly (p = 0.05) from those of unpolluted soils and streams. Anions such as nitrate (NO(-)3), sulphate (SO(2-)4) and chloride (Cl-) showed significantly (p = 0.05) higher values in the polluted soils and steams than in the unpolluted controls. The phosphate (PO(3-)4) values were lower in the polluted soils and streams than in the unpolluted controls. PO(3-)4, NO(-)3, SO(2 )4 and Cl- concentrations were generally significantly (p = 0.05) higher in soils and streams from Yorla, Zaakpon and their control than in Goi North, Goi South and Goi control. Exchangeable cations and trace metal concentrations were significantly (p = 0.05) higher in the polluted soils and streams than in the unpolluted controls. Yorla and Zaakpon polluted soils and streams had higher concentrations of the exchangeable cations and trace metals compared to values from Goi, except for Na. Results also showed significantly higher values of heavy metals (Pb, Cd, Cr and Ni) in the polluted soils compared to their controls, and values were also significantly (p = 0.05) higher in Yorla, Zaakpon and their control compared to Goi North, Goi South and Goi control. Values obtained in polluted streams were higher than in the unpolluted though not significant at the 5% level (p = 0.05). The anionic and cationic concentrations of Yorla and Zaakpon soils and streams were generally higher than those of Goi, indicating that Yorla and Zaakpon areas were more polluted than Goi. Although the concentrations of most of the anions and cations analysed in the polluted soils and streams were within the World Health Organisation's permissible values for the protection of humans and environment, the streams of Yorla, Zaakpon and Goi are in general being polluted with inorganic ions. This may make these soils and streams unacceptable for domestic and industrial uses if not treated, and soils (farmlands) may also become unsuitable for agricultural purposes. PMID- 11878649 TI - Is there a bleeding problem with platelet-active drugs? PMID- 11878650 TI - Rapacuronium: why did it fail as a replacement for succinylcholine? PMID- 11878651 TI - Can molecular similarity-activity models for intravenous general anaesthetics help explain their mechanism of action? AB - BACKGROUND: The importance of molecular shape and electrostatic potential in determining the activities of 11 structurally-diverse i.v. general anaesthetics was investigated using computational chemistry techniques. METHODS: The free plasma anaesthetic concentrations that abolished the response to noxious stimulation were obtained from the literature. The similarities in the molecular shapes and electrostatic potentials of the agents to eltanolone (the most potent anaesthetic agent in the group) were calculated using Carbo indices, and correlated with in vivo potency. RESULTS: The best model obtained was based on the similarities of the anaesthetics to two eltanolone conformers (r2=0.820). This model correctly predicted the potencies of the R- and S-enantiomers of ketamine, but identified alphaxalone as an outlier. Exclusion of alphaxalone substantially improved the activity correlation (r2=0.972). A bench mark model based on octanol/water partition coefficients (r2=0.647) failed to predict the potency order of the ketamine enantiomers. CONCLUSIONS: The results demonstrate that a single activity model can be formulated for chiral and non-chiral i.v. anaesthetic agents using molecular similarity indices. PMID- 11878652 TI - Bispectral index monitoring during electroconvulsive therapy under propofol anaesthesia. AB - BACKGROUND: The accuracy of the bispectral index (BIS) as a monitor of consciousness has not been well studied in patients who have abnormal electroencephalograms (EEG). METHODS: We studied the changes in BIS, its subparameters, and spectral entropy of the EEG during 18 electroconvulsive treatments under propofol and succinylcholine anaesthesia. A single bifrontal EEG, and second subocular channel (for eye movement estimation) was recorded. RESULTS: The median (interquartile range) BIS value at re-awakening was only 57 (47-78)--thus more than a quarter of the patients woke at BIS values of less than 50. The changes in spectral entropy values were similar: 0.84 (0.68-0.99) at the start, 0.65 (0.42-0.88) at the point of loss-of-consciousness, 0.63 (0.47-0.79) during the seizures, and 0.58 (0.31-0.85) at awakening. CONCLUSIONS: Post-ictal slow-wave activity in the EEG (acting via the SynchFastSlow subparameter) may cause low BIS values that do not correspond to the patient's clinical level of consciousness. This may be important in the interpretation of the BIS in other groups of patients who have increased delta-band power in their EEG. PMID- 11878654 TI - Comparative effect of paracetamol, NSAIDs or their combination in postoperative pain management: a qualitative review. AB - BACKGROUND: Quantitative reviews of postoperative pain management have demonstrated that the number of patients needed to treat for one patient to achieve at least 50% pain relief (NNT) is 2.7 for ibuprofen (400 mg) and 4.6 for paracetamol (1000 mg), both compared with placebo. However, direct comparisons between paracetamol and non-steroidal anti-inflammatory drugs (NSAIDs) have not been extensively reviewed. The aims of this review are (i) to compare the analgesic and adverse effects of paracetamol with those of other NSAIDs in postoperative pain, (ii) to compare the effects of combined paracetamol and NSAID with those of either drug alone, and (iii) to discuss whether the adverse effects of NSAIDs in short-term use are justified by their analgesic effects, compared with paracetamol. METHODS: Medline (1966 to January 2001) and the Cochrane Library (January 2001) were used to perform a systematic, qualitative review of postoperative pain studies comparing paracetamol (minimum 1000 mg) with NSAID in a double-blind, randomized manner. A quantitative review was not performed as too many studies of high scientific standard (27 out of 41 valid studies, including all major surgery studies) would have been excluded. RESULTS: NSAIDs were clearly more effective in dental surgery, whereas the efficacy of NSAIDs and paracetamol seemed without substantial differences in major and orthopaedic surgery, although firm conclusions could not be made because the number of studies was limited. The addition of an NSAID to paracetamol may confer additional analgesic efficacy compared with paracetamol alone, and the limited data available also suggest that paracetamol may enhance analgesia when added to an NSAID, compared with NSAIDs alone. CONCLUSION: Paracetamol is a viable alternative to the NSAIDs, especially because of the low incidence of adverse effects, and should be the preferred choice in high-risk patients. It may be appropriate to combine paracetamol with NSAIDs, but future studies are required, especially after major surgery, with specific focus on a potential increase in side-effects from their combined use. PMID- 11878653 TI - Effect of hypothermia on brain tissue oxygenation in patients with severe head injury. AB - BACKGROUND: There is renewed interest in the use of induced hypothermia as a method of neuroprotection both intraoperatively and in the intensive care management of severe brain injury. In this study we have investigated the effects of hypothermia on brain tissue oxygenation in patients with severe head injury. METHODS: Thirty patients with severe head injury (Glasgow coma score <8) were monitored with a multimodal sensor inserted into the brain which measures tissue PO2, PCO2, pH and temperature in addition to routine monitoring. Patients were cooled to a minimum of 33 degrees C when clinically indicated. RESULTS: For all 30 patients brain and systemic temperature correlated well (r=0.96). Brain temperature was consistently higher than systemic temperature by 0.41 +/- 0.26 degrees C (confidence limits). Brain tissue PO2 decreased with hypothermia, with a significant reduction below 35 degrees C (P<0.05). CONCLUSIONS: These results emphasize the advantage of measuring brain temperature directly, and suggest that decreasing brain temperature below 35 degrees C may impair brain tissue oxygenation. PMID- 11878655 TI - Rectal and parenteral paracetamol, and paracetamol in combination with NSAIDs, for postoperative analgesia. AB - BACKGROUND: We have reviewed the analgesic efficacies of rectal and parenteral paracetamol and tested the evidence for a possible additive analgesic effect of the combination of paracetamol with a non-steroidal anti-inflammatory drug (NSAID) in postoperative pain. METHODS: Randomized controlled trials were evaluated. Outcome measures were pain scores and demand for supplementary analgesia. RESULTS: Eight studies compared rectal paracetamol with placebo. One study of single-dose administration of rectal paracetamol 40-60 mg kg(-1) and three studies of repeat dosing with 14-20 mg kg(-1) showed significant analgesic efficacy, while studies of a single dose of 10-20 mg kg(-1) were negative. Ten studies compared parenteral paracetamol with placebo and eight studies showed improved pain relief with paracetamol. Of the nine studies comparing paracetamol with a combination of paracetamol and an NSAID, six studies showed improved pain relief for the combination while only two of the six studies comparing an NSAID with a combination of an NSAID and paracetamol showed improved pain relief for the combination. CONCLUSIONS: Considering the few studies available, evidence was found of a clinically relevant analgesic effect of rectal and parenteral paracetamol. Concurrent use of paracetamol and an NSAID was superior to paracetamol alone but no evidence was found of superior analgesic effect of the combination compared with the NSAID alone. PMID- 11878656 TI - Depth of sedation in children undergoing computed tomography: validity and reliability of the University of Michigan Sedation Scale (UMSS). AB - BACKGROUND: Safe care of sedated children requires ongoing assessment of the depth of sedation to permit early recognition of progression to over-sedation. This study evaluated the validity and reliability of the University of Michigan Sedation Scale (UMSS) as a measure of sedation during procedures. The UMSS is a simple observational tool that assesses the level of alertness on a five-point scale ranging from 1 (wide awake) to 5 (unarousable with deep stimulation). METHODS: Thirty-two children aged 4 months to 5 yr (mean 1.5 yr), sedated for computed tomography (CT), were studied prospectively. The CT nurse assessed sedation using the UMSS before sedative administration and every 10 min thereafter. The child was videotaped during each assessment, and segments were edited and their order was randomized. Four nurses blinded to sedative administration viewed the segments and scored sedation using the UMSS. One of these nurses also scored sedation using a visual analogue scale (VAS) and another using the Observer's Assessment of Alertness/Sedation Scale (OAAS). To examine the test-retest reliability, 75 randomly selected video segments were viewed and scored on a second occasion. RESULTS: Changes in scores from baseline to discharge supported construct validity (P<0.0001). Criterion validity was demonstrated by significant correlations between the UMSS and the VAS and OAAS. There was good interobserver agreement between blinded observers' scores for each level of sedation and at discharge, and between blinded observers and the CT nurse for scores of 0 and 1 (lighter levels of sedation), but less agreement for scores 2 and 3 (deeper sedation) and discharge scores. Test-retest reliability was supported by agreement in the observers' UMSS scores. CONCLUSION: The UMSS is a simple, valid and reliable tool that facilitates rapid and frequent assessment and documentation of depth of sedation in children. PMID- 11878657 TI - Positive pressure ventilation during fibreoptic intubation: comparison of the laryngeal mask airway, intubating laryngeal mask and endoscopy mask techniques. AB - BACKGROUND: The efficacy of delivery of mechanical ventilation through different airway devices during fibreoptic intubation is not known. METHODS: We compared the laryngeal mask airway (LMA), intubating laryngeal mask (ILM) and endoscopy mask for positive pressure ventilation (PPV) during fibreoptic intubation. In 80 adult paralysed patients, fibreoptic intubation was performed during PPV using a combination of a size 3 or 4 LMA with a 6.0 mm nasal RAE tracheal tube (LMA3/4 group; n=22), a size 5 LMA with a 7.0 mm nasal RAE tube (LMA5 group; n=18), an ILM with an 8.0 mm special reinforced tracheal tube (ILM group; n=20) or an endoscopy mask (Patil mask) with a 7.5 mm standard tracheal tube (Patil group; n=20). The inspiratory and expiratory tidal volumes (VI and VE) with a ventilation pressure of 20 cm H2O were measured using a pneumotachograph. RESULTS: Mean VE values during fibreoptic intubation in the LMA5 [5.3 (SD 1.5) ml kg(-1)] and ILM [7.1 (2.3) ml kg(-1)] groups were greater than in the LMA3/4 group [2.6 (1.0) ml kg(-1), P<0.0001]. The mean VE was greater in the Patil group [20.6 (4.9) ml kg(-1)] than in the other three groups (P<0.0001). Gastric insufflation during intubation was more frequent in the Patil group (30%) than in the other three groups (4.5-5.6%) (P<0.05). CONCLUSION: PPV is possible with the LMA, ILM or endoscopy mask during fibreoptic intubation. With an airway pressure of 20 cm H2O, ventilation during intubation using a size 3 or 4 LMA was almost insufficient, while ventilation using a size 5 LMA or an ILM was almost acceptable. Ventilation during intubation with the endoscopy mask was greater than that with the LMA or ILM, but gastric insufflation was more frequent. PMID- 11878658 TI - Xenon produces minimal haemodynamic effects in rabbits with chronically compromised left ventricular function. AB - BACKGROUND: Xenon has only minimal haemodynamic side-effects on normal myocardium and might be a preferable anaesthetic agent for patients with heart failure. We studied the haemodynamic changes caused by 70% xenon in rabbits with chronically compromised left ventricular (LV) function. METHODS: Anaesthetized rabbits were thoracotomized and a major coronary artery was ligated to induce ischaemic heart disease. Nine weeks later, rabbits were again anaesthetized (ketamine/propofol), and haemodynamics were measured during inhalation of 70% xenon using echocardiography [LV end-diastolic dimension (LVedD), fractional shortening (FS), velocity of circumferential fibre shortening (VcF), ejection fraction (EF)] in closed-chest animals. Subsequently, rabbits were thoracotomized and instrumented for measurement of LV pressure (tip manometer), LV dP/dtmax and cardiac output (ultrasonic flow probe). Haemodynamics were recorded again during inhalation of 70% xenon. RESULTS: All rabbits had compromised LV function 9 weeks after coronary artery ligation. Mean LVedD increased from 12.9 (SD 0.9) mm to 17.1 (0.4) mm; EF decreased from 73 (9) to 64 (8)%; FS decreased from 36 (7) to 29 (5)%; VcF decreased from 28.9 (6.8) to 17.6 (4.7) mm s(-1); all P<0.05. Inhalation of 70% xenon had no effect on haemodynamics in closed-chest rabbits, as measured by echocardiography. After invasive instrumentation, small decreases in LV pressure from 78 (20) to 72 (19) mm Hg, LV dP/dtmax from 3081 (592) to 2633 (503) mm Hg s(-1) and cardiac output from 239 (69) to 225 (71) ml min(-1) were observed during xenon inhalation (all P<0.05). CONCLUSION: These data show that xenon has only minimal negative inotropic effects in rabbits with LV dysfunction after coronary artery ligation. PMID- 11878659 TI - Analgesia for pelvic brachytherapy. AB - Pelvic brachytherapy presents the anaesthetist with numerous challenges. Patients vary from highly distressed young adults, to the elderly with coincidental disease severe enough to preclude surgery. The painful radioactive implants remain in place for a number of days. Treatment in isolated rooms reduces radiation exposure to staff, but makes close postoperative monitoring difficult, so the analgesic technique should involve minimum risk to the patient. Although there is very little published evidence of specific analgesic techniques in this area, knowledge of these problems allows the anaesthetist to select appropriate systemic analgesics and regional blocks to provide safe and effective pain relief. PMID- 11878660 TI - Effects of temperature and haematocrit on the relationships between blood flow velocity and blood flow in a vessel of fixed diameter. AB - BACKGROUND: To determine whether temperature and haematocrit (Hct) alter the relationship between blood flow (BF) and blood flow velocity (BFV). METHODS: Using a transcranial Doppler apparatus, we measured the peak velocity of whole blood cells pumped by a cardiopulmonary bypass (CPB) circuit, through a 0.15-cm internal diameter segment of rigid tubing. BF and BFV relationships were obtained at temperatures of 19, 28, and 37 degrees C and at Hct of 0.05, 0.22, 0.39, and 0.54, by altering CPB flow over a range from 10 to 100 cc/min. Linear regression analysis was performed. RESULTS: The relationship between velocity and flow for the pooled Hct data was y=(0.43)x+0.86, r2=0.998 and 95% CI (0.999-1) whereas the association for the temperature data was y=(0.42)x+0.02, r2=0.9998 and 95% CI (0.999-0.9997). Changes of blood viscosity had no effect on velocity at a given flow rate. The combined effect of Hct and temperature on velocity for the relationship with flow is expressed by: y=1.3+2.4x. CONCLUSION: In fixed diameter vessels with laminar flow, the linear relationship between flow and velocity is not affected by changes in temperature and Hct in clinical ranges. These results are explained by the Fahraeus-Lindquist effect. They support the use of transcranial Doppler sonography to estimate cerebral blood flow in infants who may have large variations of Hct and/or temperature during bypass. PMID- 11878661 TI - Improvement of information gained from the pre-anaesthetic visit through a quality-assurance programme. AB - BACKGROUND: Pre-anaesthetic evaluation is a fundamental component of anaesthetic practice. The aims of the present study were to assess the quality of the preoperative anaesthetic information gathered and to observe the quality profile after the introduction of a standardized form. This occurred through a four-step quality assurance programme over a 4-yr period. METHODS: The proportion of cases in which a complete recording of data was collected at the preoperative assessment was evaluated by searching in each patient's medical record for what was considered to be the minimum information required to provide satisfactory perioperative care. Fifteen criteria were selected. The recovery profile for each indicator and a global quality index (GQI expressed in %) for each patient's record were collected. In phase 1, the existing situation was assessed. Next, a standardized pre-anaesthetic form (PAF 1) was designed and its implementation evaluated (Phase 2). Phase 3 was performed 16 months after implementation of PAF 1 to assess the long-term value. The form was revised (PAF 2) and its use evaluated again 6 months later (Phase 4). For each evaluation, the records of a 1 month period were examined. Overall 1129 medical records were audited. RESULTS: The GQI increased significantly from 62 to 88% with similar changes for both elective and emergency cases. The recovery profile was improved for most indicators. CONCLUSIONS: We conclude that the quality of information recorded from the pre-anaesthetic visit is improved by using a standardized form. This will hopefully help to improve patient outcome and facilitate computerization of the anaesthetic record. PMID- 11878662 TI - How reliable is length-based determination of body weight and tracheal tube size in the paediatric age group? The Broselow tape reconsidered. AB - BACKGROUND: The Broselow tape was designed to estimate body weight and tracheal tube size on the basis of the body length of emergency paediatric patients. The tape was validated previously in US populations. We assessed its accuracy in a sample of European children by reviewing paediatric anaesthetic charts at the Triemli City Hospital for 1999. METHODS: Age, body length and body weight measured before surgery as well as the size of the tracheal tube used were recorded. The body weight was estimated on the basis of body length using the Broselow tape and was compared with the measured weight. Tracheal tube size selections using the Broselow tape and an age-based formula were compared with the size of the tube used. RESULTS: A good correlation was found between the Broselow weight and the measured weight (r2=0.88). Bland-Altman analysis revealed a mean bias of -0.52 kg for the entire study population. For children < or = 20 kg the mean bias was -0.05 kg, and for children > 20 kg was -1.05 kg. The Broselow weight was found to be within a 10% error of the measured weight in 65% of children. Tracheal tube selection by the Broselow tape method was adequate in 55% but underestimated the actual tube size in 39%. The age-based formula matched the actual tracheal tube size in 41% of children but overestimated it in 57%. CONCLUSIONS: The Broselow tape is an accurate means to assess body weight from length in smaller children; in older children it underestimated body weight. Endotracheal tube size selection by the Broselow tape appears to match the size of the tube used better than the age-based formula. The results in a European sample of children are comparable to the US data. PMID- 11878663 TI - Effect of spinal anaesthesia on plasma concentrations of glutathione S transferase. AB - BACKGROUND: Plasma glutathione S-transferase (GST) concentration measurement is a sensitive and specific index of hepatocellular injury. GST concentration increases after anaesthesia with most volatile anaesthetic agents, but not after propofol. Such increases are thought to result from reduced liver blood flow. The effect on GST concentration of spinal (subarachnoid) anaesthesia, which might also reduce liver blood flow, is not known. METHODS: We studied the effects of spinal anaesthesia on GST concentrations measured by specific radioimmunoassay in 33 patients undergoing intermediate orthopaedic, general or gynaecological surgery. GST concentrations were measured before anaesthesia and 3, 6 and 24 h after induction of anaesthesia. Hypotension (systolic blood pressure <70% of pre induction value) was rapidly corrected with i.v. ephedrine. RESULTS: Mean duration of surgery was 41 min (range 11-80). No increase in GST concentration was observed at any time, but at 24 h GST concentration was significantly reduced (P<0.05). One patient in whom hypotension was not treated developed a greatly increased GST concentration at 3 h. CONCLUSION: We found no association between spinal anaesthesia and disturbance of hepatocellular integrity when hypotension does not occur or is rapidly corrected. PMID- 11878664 TI - Management of life-threatening haemoptysis. AB - Massive haemoptysis represents a major medical emergency that is associated with a high mortality. Here we present two cases of life-threatening haemoptysis, the first caused by rupture of an aortic aneurysm into the lung in a 37-yr-old woman with polyarteritis nodosa and the second caused by massive bleeding from an angiectatic vascular malformation in the right main bronchus in a 21-yr-old woman. Fibreoptic bronchoscopy played an essential role in the diagnostic process and management of the respiratory tract. Diagnosis in the first case was obtained by CT scan and the aneurysm was treated surgically. In the second case, bronchial arteriography contributed to both definitive diagnosis and treatment. Initial cardiorespiratory management, diagnostic procedures and definitive therapy are described and reviewed. Adequate early management of the cardiorespiratory system is essential to the outcome. Aggressive measures to elucidate the cause of haemoptysis and prompt therapy are warranted because of the high risk of recurrence. PMID- 11878665 TI - Mechanical closure of the vocal cords with the laryngeal mask airway ProSeal. AB - We report a case of mechanical closure of the vocal cords with the laryngeal mask airway ProSeal (PLMA) in an anaesthetized, paralysed 59-yr-old, 88 kg female undergoing lower-limb vascular surgery. Near-complete airway obstruction developed after a size-5 PLMA was inserted and the cuff inflated with 20 ml of air. Fibreoptic inspection revealed that the PLMA was correctly positioned, but the vocal cords were closed. Withdrawal of air from the cuff and/or moving the head and neck into the sniffing position resolved this problem. PMID- 11878666 TI - Infected mediastinitis secondary to perforation of superior vena cava by a central venous catheter. AB - We describe the first case of infected mediastinitis associated with central venous catheter insertion. The rare occurrence of this complication may be explained by the fact that it results from central venous catheter-related bloodstream infection and catheter perforation of superior vena cava. The symptoms of this complication (chest pain, dyspnoea) are not specific. Diagnosis should be confirmed by chest x-ray and computerized tomography which show hydromediastinum and pleural effusion. Removal and subsequent culture of the catheter tip will confirm infection. Appropriate antibiotic therapy, guided by sensitivities of the cultured organisms, should be commenced. Any pleural effusion should be drained by thoracocentesis, and the pleural fluid cultured. In case of fever, bacteraemia or shock, a thoracotomy to drain mediastinal and pleural effusions may be considered. PMID- 11878667 TI - Do ex-premature infants need mechanical ventilation for inguinal hernia repair? PMID- 11878668 TI - Downward movement of syringe pumps reduces syringe output. PMID- 11878669 TI - Sedative and analgesic practice in the intensive care unit. PMID- 11878670 TI - Allergy to chlorhexidine-coated central venous catheters revisited. PMID- 11878671 TI - Outcome measures. PMID- 11878672 TI - The alveolar surface and the geodesic planes hypothesis. PMID- 11878673 TI - Informed consent for regional anaesthesia. PMID- 11878674 TI - Is it safe to artificially ventilate a paralysed patient through a laryngeal mask? PMID- 11878675 TI - Thoughts on the channel size of the motor endplate acetylcholine receptor. PMID- 11878676 TI - Pulse oximetry, carbon monoxide and protective ventilatory strategies in severe ARDS. PMID- 11878678 TI - Central noradrenergic neurones and the mechanism of general anaesthesia. PMID- 11878677 TI - Failed intubation due to posterior fossa haematoma requiring emergency percutaneous tracheostomy. PMID- 11878679 TI - Comparative mortality in anaesthesia. PMID- 11878680 TI - PCA: patient-controlled analgesia or politically correct analgesia? PMID- 11878681 TI - Cardioventilatory coupling in heart rate variability: the value of standard analytical techniques. AB - In a group of spontaneously breathing anaesthetized subjects, we examined the ability of simple spectral and non-linear methods to detect the presence of cardioventilatory coupling in heart rate time series. Using the proportional Shannon entropy (H(RI-1)) of the RI(-1) interval (interval between inspiration and the preceding ECG R wave) as a measure of coupling, we found no correlation between H(RI-1) and either the fractal dimension or approximate entropy of the heart rate time series. We also observed no difference in the distribution of heart rate variability (HRV) spectral power in three frequency ranges (high, 0.15 0.45 Hz; low, 0.08-0.15 Hz; very low, 0.02-0.08 Hz) between uncoupled epochs and coupling patterns I, III and IV. Because of its association with low breathing frequencies, pattern II coupling epochs showed exaggerated low-frequency power as the high-frequency 'respiratory' peak fell into the low-frequency range. We conclude that coupling pattern is largely independent of autonomic tone and that these standard methods of HRV analysis are limited in their ability to detect the presence of cardioventilatory coupling in heart rate time series. PMID- 11878682 TI - Cardioventilatory coupling in heart rate variability: methods for qualitative and quantitative determination. AB - In this study we sought to develop quantitative methods for determining the presence of cardioventilatory coupling in raw heart rate time series. The beat-to beat RR interval time series of 98 anaesthetized, spontaneously breathing subjects were represented graphically as (1) raw RR interval time series, (2) RR consecutive difference time series and (3) a phase portrait of the RR consecutive difference time series. We then examined the relationships between the presence of cardioventilatory coupling in these epochs and the plot appearance and entropy measures derived from these plots. We observed that coupling was significantly associated with the presence of banding in the raw heart rate and RR consecutive difference time series, and with discrete clustering within the RR consecutive difference phase portrait. A significant correlation was found between coupling and the entropy of the RR consecutive difference time series and its phase portrait. We conclude that, with some provisos, these simple graphical and derived quantitative measures provide a basis for the determination of cardioventilatory coupling from heart rate time series. PMID- 11878683 TI - Volume kinetics of glucose solutions given by intravenous infusion. AB - Glucose solutions given by intravenous (i.v.) infusion exert volume effects that are governed by the amount of fluid administered and also by the metabolism of the glucose. To understand better how the body handles glucose solutions, two volume kinetic models were developed in which consideration was given to the osmotic fluid shifts that accompany the metabolism of glucose. These models were fitted to data obtained when 21 volunteers who were given approximately 1 litre of glucose 2.5 or 5% or Ringer's solution (control) over 45 min. The maximum haemodilution was similar for all three fluids, but it decreased more rapidly when glucose had been infused. The volume of distribution for the infused glucose molecules was larger (approximately 12 litres) than for the infused fluid, which amounted to (mean (SEM)) 3.7 (0.3) (glucose 2.5%), 2.8 (0.2) (glucose 5%), and 2.5 (0.2) litres (Ringer). Fluid accumulated in a remote (cellular) body fluid space when glucose had been administered (approximately 0.2 and 0.4 litres, respectively), while expansion of an intermediate fluid space (7.1 (1.3) litres) could be demonstrated in 33% of the Ringer experiments. In conclusion, kinetic models were developed which consider the relationship between the glucose metabolism and the disposition of intravenous fluid. One of them, in which infused fluid expands two instead of three body fluid spaces, was successfully fitted to data on blood glucose and blood haemoglobin obtained during infusions of 2.5 and 5% glucose. PMID- 11878684 TI - Heparin pretreatment does not alter heparin requirements during cardiopulmonary bypass. AB - Heparin infusion may cause heparin resistance and may affect monitoring by measurement of the activated coagulation time (ACT), making the assessment of anticoagulation difficult, with the risk of over- or undertreatment, especially during cardiac surgery. We studied two groups of patients undergoing cardiopulmonary bypass (CPB): patients on heparin infusions (group H) and heparin naive controls (group C). All patients received heparin 300 IU kg(-1) before CPB and a further dose of 5000 IU if the ACT 5 min after commencing bypass was less than 400 s. Measurements of ACT, heparin concentration, antithrombin-3, thrombin antithrombin complex, prothrombin fragment F(1+2) and D-dimers were made before and 5 and 20 min after start of CPB. A second dose of heparin was given to eight out of 18 patients in group C and 10 out of 24 in group H. Antithrombin-3 in group H was significantly less than in group C at 5 min [59 (14) vs 52 (9)%, P<0.05]. ACT was significantly lower in group H than group C at 20 min [387 (64) vs 431 (67) s, P<0.05]. Despite ACTs of less than 400 s in both groups, no coagulation was seen, suggesting that 300 IU kg(-1) heparin is a safe dose for anticoagulation in CPB even after heparin therapy. PMID- 11878685 TI - Safety of oral nicorandil before coronary artery bypass graft surgery. AB - Nicorandil is a K(ATP) channel opener used to treat angina. It is cardioprotective and a vasodilator. We conducted a prospective, randomized, double-blind, placebo-controlled study to assess oral nicorandil in patients undergoing coronary artery bypass grafting (CABG) with cardiopulmonary bypass (CPB). Twenty-two patients received nicorandil (10 mg twice a day) and 23 patients received placebo. Haemodynamic data were recorded before induction of anaesthesia (T0), 5 and 20 min after starting mechanical ventilation (T1, T2), before aortic cannulation (T3), after 30 min of CPB (T4), 10 min after CPB (T5) and after 3, 8 and 18 h in the intensive care unit (T6, T7, T8). Serum proteins (creatine kinase metabolite and cardiac troponin I) were measured before and 8 and 18 h after surgery. Haemodynamic values did not differ between the two groups. There was no tachycardia during the study, no significant difference in hypotensive episodes, ST segment changes and no changes in cardiac enzymes. Myocardial infarction after surgery was similar in the two groups. Vasoactive therapy was similar in the two groups. Nicorandil can be continued safely up to premedication without deleterious haemodynamic consequences, but a myocardial protective effect of nicorandil in CABG surgery was not found. PMID- 11878686 TI - Postoperative changes in visual evoked potentials and cognitive function tests following sevoflurane anaesthesia. AB - We tested the hypothesis that minor disturbance of the visual pathway persists following general anaesthesia even when clinical discharge criteria are met. To test this, we measured visual evoked potentials (VEPs) in 13 ASA I or II patients who did not receive any pre-anaesthetic medication and underwent sevoflurane anaesthesia. VEPs were recorded on four occasions, before anaesthesia and at 30, 60, and 90 min after emergence from anaesthesia. Patients completed visual analogue scales (VAS) for sedation and anxiety, a Trieger Dot Test (TDT) and a Digit Symbol Substitution Test (DSST) immediately before each VEP recording. These results were compared using Student's t-test. P<0.05 was considered significant. VEP latency was prolonged (P<0.001) and amplitude diminished (P<0.05) at 30, 60, and 90 min after emergence from anaesthesia, when VAS scores for sedation and anxiety, TDT, and DSST had returned to pre-anaesthetic levels. PMID- 11878687 TI - Influence of oral tramadol on the dynamic ventilatory response to carbon dioxide in healthy volunteers. AB - We tested the effect of tramadol on ventilatory control by quantifying its effect on the steady-state ventilatory carbon dioxide response and by locating its site of respiratory action within the ventilatory control system. We imposed square wave changes in end-tidal carbon dioxide (approximately 1 kPa; end-tidal oxygen concentration kept constant at resting levels) in 10 healthy volunteers (six men, four women) before and after oral ingestion of 100 mg tramadol, and measured the ventilatory responses. Each hypercapnic response was separated into a fast, peripheral and a slow, central component. Two control and two tramadol carbon dioxide studies were performed in each subject. Tramadol reduced the total ventilatory carbon dioxide sensitivity by approximately 30% from 12.8 (6.1) [lower (25%) and upper (75%) quartiles 7.4 and 16.6 litre min(-1) kPa(-1)] to 9.1 (5.3) (5.3-14.1) litre min(-1) kPa(-1) (P<0.001). The fast and slow response gains were reduced by 23 (46) (3-54)% (P<0.05) and 30 (22) (15-54)% (P<0.01) respectively. The ratio of these carbon dioxide sensitivities and the apnoeic threshold were not significantly changed by tramadol. We suggest that tramadol affects the ventilatory control system by acting at the mu-opioid receptors in the respiratory integrating centres within the brainstem. PMID- 11878688 TI - No clinical evidence of acute opioid tolerance after remifentanil-based anaesthesia. AB - We have prospectively assessed whether remifentanil-based anaesthesia is associated with clinically relevant acute opioid tolerance, expressed as greater postoperative pain scores or morphine consumption. Sixty patients undergoing elective gynaecological, non-laparoscopic, surgery were randomly assigned to receive remifentanil (group R, n=30) or sevoflurane (group S, n=30) based anaesthesia. Postoperative analgesia was provided with morphine through a patient controlled infusion device. Mean (SD) remifentanil infusion rate in group R was 0.23 (0.10) microg kg(-1) min(-1) and mean inspired fraction of sevoflurane in group S was 1.75 (0.70)%. Mean (SD) cumulative morphine consumption during the first 24 postoperative hours was similar between groups: 28.0 (14.2) mg (group R) vs 28.6 (12.4) mg (group S). Pain scores, were also similar between groups during this period. These data do not support the development of acute opioid tolerance after remifentanil-based anaesthesia in this type of surgery. PMID- 11878689 TI - Transtracheal high frequency jet ventilation for endoscopic airway surgery: a multicentre study. AB - Serious complications during high frequency jet ventilation (HFJV) are rare and have been documented in animals and in case reports or short series of patients with a difficult airway. We report complications of transtracheal HFFJV in a prospective multicentre study of 643 patients having laryngoscopy or laryngeal laser surgery. A transtracheal catheter could not be inserted in two patients (0.3%). Subcutaneous emphysema (8.4%) was more frequent after multiple tracheal punctures. There were seven pneumothoraces (1%), two after laser damage to the injector, one after difficult laryngoscopy, four with no clear cause. Arterial desaturation of oxygen was more frequent during laser surgery and in overweight patients. Transtracheal ventilation from a ventilator with an automatic cut-off device is a reliable method for experienced users. Control of airway pressure does not prevent a low frequency of pneumothorax. PMID- 11878690 TI - Tracheal tube pressure change during magnetic stimulation of the phrenic nerves as an indicator of diaphragm strength on the intensive care unit. AB - Diaphragm strength can be assessed from twitch gastric (TwPgas), twitch oesophageal (TwPoes), and twitch transdiaphragmatic pressure (TwPdi) in response to phrenic nerve stimulation. This requires the passage of balloon catheters, which may be difficult. Changes in pressure measured at the mouth during phrenic nerve stimulation avoid the need for balloon catheters. We hypothesized that pressures measured at the tracheal tube during phrenic stimulation, could also reflect oesophageal pressure change as a result of isolated diaphragmatic contraction and, therefore, reflect diaphragm strength. We aimed to establish the relationship between twitch tracheal tube pressure (TwPet), TwPoes, and TwPdi in patients in the supine and sitting positions. The phrenic nerves were stimulated magnetically bilaterally, in 14 ICU patients while supine and on another occasion while sitting up at 45 degrees. In the sitting position mean TwPoes was 9.1 cm H2O and TwPet 11.3 cm H2O (mean(SD) difference -2.2 (SD 1.5)). In the supine position mean TwPoes was 8.1 cm H2O and TwPet 9.9 cm H2O (mean difference -1.8 (2.2)). The difference between TwPoes and TwPet was less at low twitch amplitude; less than +/- 1 cm H2O below a mean twitch height of 8 cm H2O supine and 10 cm H2O sitting. Sitting TwPet was related to TwPoes r2=0.93 and TwPdi r2=0.65 (P<0.01). Supine TwPet was related to TwPoes r2=0.84 and TwPdi r2=0.83 (P<0.01). The mean within occasion coefficient of variation while sitting was TwPet=13.3%, TwPoes=13.9%, TwPdi=11.2%, and supine TwPet=11.6%, TwPoes=14.6%, TwPdi=11.8%. We conclude that TwPet reflects TwPoes during diaphragmatic stimulation and is worthy of further study to establish its place as a guide to the presence of respiratory muscle strength and fatigue. PMID- 11878691 TI - Difference in sensitivity to vecuronium between patients with ocular and generalized myasthenia gravis. AB - Patients with myasthenia gravis show sensitivity to non-depolarizing neuromuscular blocking drugs, but little is known about differences in this sensitivity between types of myasthenia. In 10 patients with ocular myasthenia gravis and 10 with generalized myasthenia gravis, twitch tension was monitored in the adductor pollicis muscle by supramaximal train-of-four stimulation of the ulnar nerve during anaesthesia with sevoflurane 2.5% and nitrous oxide 60%. After baseline measurement, an initial dose of vecuronium 10 microg kg(-1) was given. When the twitch height stabilized (maximum block after the first 10 microg kg( 1)), the next incremental dose of 10 microg kg(-1) was given and repeated until block, defined as [1-(first twitch/baseline first twitch)] x 100 reached 90%. Maximum block after the first dose of vecuronium in ocular patients was significantly less than that in generalized patients (median 51 vs 91%; P<0.05). Onset of block after the first dose of vecuronium was significantly slower in ocular than in generalized myasthenic patients (mean 300 vs 200 s; P<0.05). Doses required to attain a block of 90% or more were significantly higher in ocular than in generalized patients (median 20 vs 10 microg kg(-1); P<0.05). Clinicians should consider the type of disease according to the Osserman classification when using non-depolarizing neuromuscular. blocking drugs in patients with myasthenia gravis. PMID- 11878692 TI - Platelet function during pregnancy: an evaluation using the PFA-100 analyser. AB - In clinical practice, the only tests of platelet function are bleeding time and platelet number. Bleeding time lacks sensitivity and specificity but the PFA-100, an in vitro analyser of platelet function may be of value. This study aimed to evaluate any correlation between platelet number and function using the PFA-100 in pregnant women. During a 21-month period, platelet function was evaluated in whole blood as part of the pre-anaesthetic coagulation testing screen with the PFA-100 using collagen and epinephrine (PFA-EPI) or ADP (PFA-ADP) as platelet agonists. Thrombocytopenia was defined as a platelet number less than 150 G litre(-1). The patients were divided into four groups: Group I (n=110) normal pregnancy; Group II (n=38) thrombocytopenia of pregnancy; Group III (n=13) women with pre-eclampsia without thrombocytopenia; Group IV (n=19) women with pre eclampsia and thrombocytopenia. Results are expressed as mean (SD). Platelet count was not statistically different between Groups II and IV (111.1 (23.1) vs 99.5 (28.0) G litre(-1)). PFA-EPI was statistically increased in Group II (124.0 (26.3) s), Group III (128.3 (17.9) s), and Group IV (143.6 (47.7) s) compared with normal pregnant patients (114.6 (27.3) s, P<0.05, Mann-Whitney U-test). PFA ADP was statistically increased only in Group II compared with normal pregnant patients (90.5 (18.9) vs 80.2 (11.2) s, P<0.05). PFA values were increased above normal laboratory values in (four of 38) Group II patients and (six of 19) Group IV patients but in no patients in Group III. PFA-ADP results were correlated with platelet count only in Group IV (r=-0.74, P=0.0003). The increased PFA values and the correlation between PFA-ADP and platelet number in hypertensive thrombocytopenic women confirms that platelet function may be decreased in such patients. In patients with pregnancy-induced thrombocytopenia, platelet function may be preserved when the platelet count is as low as 60 G litre(-1). PMID- 11878693 TI - Diffusion of nitrous oxide into the pleural cavity. AB - We postulated that nitrous oxide transfer into the pleural cavity can occur by diffusion from the alveoli, independent of vascular transport. Under general anaesthesia, six sheep were studied in two phases, a control and an experimental phase. The sheep were anaesthetized, intubated, and received positive pressure mechanical ventilation. A catheter was placed in the right pleural cavity and 150 ml air injected. The animals were ventilated with 100% oxygen. The inspired gas was changed to a mixture of 50% nitrous oxide and 50% oxygen, and the rate of increase of nitrous oxide concentration in the pleural space was measured. The animals were then ventilated with 100% oxygen and then killed by exsanguination while ventilation was continued. The inspired mixture was changed to 50% nitrous oxide and 50% oxygen and the rate of increase in nitrous oxide concentration was measured in the pleural space again. During venitilation with nitrous oxide in the living animals, the concentration of nitrous oxide in the pleural cavity increased rapidly and decreased to zero during ventilation with 100% oxygen. During ventilation without circulation, the rate of increase in the concentration of nitrous oxide in the pleural cavity was the same as in the control phase. This suggests that nitrous oxide enters the pleural space by diffusion, rather than by vascular delivery. This mechanism may explain the rapid increase in the volume of pneumothorax if nitrous oxide is given in the inspired gas. PMID- 11878694 TI - Inhibition of active sodium absorption leads to a net liquid secretion into in vivo rabbit lung at two levels of alveolar hypoxia. AB - Active sodium transport across alveolar epithelium is known to contribute to the resolution of pulmonary oedema. We have attempted to assess whether sodium transport is essential to prevent liquid accumulation in healthy pulmonary alveoli exposed to mild hypoxia, and whether its contribution to liquid absorption differs between mild and moderate levels of hypoxia. In twenty-four anaesthetized adult rabbits we used direct bronchial cannulation to measure liquid movement from the liquid-filled left lung over 3.5 h. Half of the rabbits were studied at a level of mixed venous (and alveolar) oxygen partial pressure, PVO2, of 6.5 kPa and half at 4.5 kPa. PVO2 was altered by changing the inspired oxygen fraction in the ventilated right lung. Alveolar hydrostatic pressure was 0.3 kPa. In each group of 12, six animals with inhibitors of sodium transport in the isosmotic instillate were compared with six controls. We have shown an alveolar liquid secretion (approximately 0.6 microl min(-1) (kg body weight)(-1)) in the presence of inhibitors of active transport and an absorption (approximately 4 microl min(-1) (kg body weight)(-1)) in controls. Changing PVO2 had no influence on these movements. We conclude that, in this model of pulmonary oedema, active sodium transport appears to be essential for prevention of alveolar liquid accumulation via secretion. Furthermore, the contribution of active sodium transport to liquid absorption remains constant at oxygen tensions between 4.5 and 6.5 kPa. PMID- 11878696 TI - The new neuromuscular blocking agents: do they offer any advantages? AB - The pharmacodynamics and pharmacokinetics of the two most recent aminosteroid neuromuscular blocking drugs to become available, rapacuronium bromide (Org 9487) and rocuronium bromide are reviewed. Two new classes of drug with neuromuscular blocking properties, the bis-tetrahydroisoquinolinium chlorofumarates and the tropinyl diester derivatives are introduced. Comparisons between these drugs and mivacurium and cisatracurium are made. Rapacuronium 1.5 mg kg(-1) (ED95 1 mg kg( 1)), produces maximal neuromuscular block in 54 s. Time to recovery of the train of-four ratio to 0.7 is achieved within 20 min after neostigmiine 0.05 mg kg(-1) given at 2 min. The plasma clearance of rapacuronium is 7-8 ml kg(-1) min(-1). Rapacuronium undergoes hepatic metabolism: no prolongation of effect has been reported after a single bolus or a short infusion in patients with hepatic or renal failure. Org 9488 is the 3-desacetyl metabolite of rapacuronium, which has neuromuscular blocking properties. Its much lower clearance (1.28 ml kg(-1) min( 1)) and plasma equilibration constant (0.105 min(-1)) may limit the prolonged use of rapacuronium. Rocuronium given at 2xED95 produces maximal neuromuscular block in 1 min. Spontaneous recovery of the train-of-four ratio to 0.7 takes over 40 min. Rocuronium has a plasma clearance of 4 ml kg(-1) min(-1). Its pharmacodynamics are altered in hepatic and renal disease. A number of anaphylactoid reactions to rocuronium have been reported recently. The bis tetrahydroisoquinolinium chlorofumarate GW280430A has an ED95 of 0.19 mg kg(-1). Given at three times this dose, onset of neuromuscular block occurs within 100 s; the duration of block is 8-9 min. Following a 2 h infusion, the recovery index does not seem to be increased. Early studies suggest that this drug has no adverse cardiovascular or respiratory side-effects. The tropinyl diester derivative G-1-64 will produce 80-90% neuromuscular block in less than 2 min using 3xED80. Ninety per cent recovery of the first twitch of the train-of-four occurs after 5-7 min using one ED80. A recovery index of less than 2 min has been reported in rats. All the tropinyl diesters appear to produce vagal block. PMID- 11878695 TI - One MAC of sevoflurane provides protection against reperfusion injury in the rat heart in vivo. AB - Volatile anaesthetics protect the heart against reperfusion injury. We investigated whether the cardioprotection induced by sevoflurane against myocardial reperfusion injury was concentration-dependent. Fifty-eight alpha chloralose anaesthetized rats were subjected to 25 min of coronary artery occlusion followed by 90 min of reperfusion. Sevoflurane was administered for the first 15 min of reperfusion at concentrations corresponding to 0.75 (n=11), 1.0 (n=11), 1.5 (n=13), or 2.0 MAC (n=12). Eleven rats served as untreated controls. Left ventricular peak systolic pressure (LVPSP, tipmanometer) and cardiac output (CO, flowprobe) was measured. Infarct size (IS, triphenyltetrazolium staining) was determined as percentage of the area at risk. Baseline LVPSP was 131 (126 135) mm Hg (mean (95% confidence interval)) and CO 33 (31-36) ml min(-1), similar in all groups. During early reperfusion, sevoflurane reduced LVPSP in a concentration-dependent manner to 78 (67-89)% of baseline at 0.75 MAC (not significant vs controls 99 (86-112)%), 71 (62-80)% at 1 MAC (P<0.05), 66 (49-83)% at 1.5 MAC (P<0.05) and 56 (47-65)% at 2 MAC (P<0.05). CO remained constant. While 0.75 MAC of sevoflurane had no effect on IS (34 (27-41)% of the area at risk) compared with controls (38 (31-45)%, P=0.83), 1.0 MAC reduced IS markedly to 23 (17-30)% (P<0.05). Increasing the concentration to 1.5 MAC (23 (17-30)%) and 2 MAC (23 (13-32)%, both P<0.05 vs controls) had no additional protective effect. One MAC sevoflurane protected against myocardial reperfusion injury. Increasing the sevoflurane concentration above 1 MAC resulted in no further protection. PMID- 11878697 TI - Thrombocytosis in intensive care. AB - We conducted a retrospective study of platelet count in 226 patients admitted for critical care over a 5-month period, to explore the incidence of thrombocytosis and its relation to admission category, duration of ICU stay and outcome. Our findings indicate that thrombocytosis is not rare in ICU patients. At least one platelet count greater than 450x10(9) litre(-1) was found in 21.7% of patients and was associated with lower ICU mortality (P=0.003), lower hospital mortality (P=0.006), but longer duration of ICU stay (P<0.0001). Thrombocytosis may serve as an independent predictor of favourable outcome in ICU patients. PMID- 11878698 TI - Preoxygenation: a comparison of three different breathing systems. AB - An end-tidal expiratory oxygen concentration (FE'O2) greater than 0.90 is considered to be adequate for preoxygenation. This is generally achieved using a face mask, but this can be unsatisfactory in some patients. We compared preoxygenation in 30 healthy volunteers using a face mask, the NasOral system, which is a novel preoxygenation device, and a mouthpiece with a nose-clip. We measured the maximal FE'O2, the FE'O2 after 2 min and the time to reach maximal FE'O2 and recorded the subjective judgement of the volunteers. The maximal FE'O2 with face mask and mouthpiece was significantly greater than with the modified NasOral system (P<0.05 and P<0.01). With the former devices, a FE'O2 of 0.90 was achieved in 73% of the volunteers vs 46% with the modified NasOral system. Using the mouthpiece, the FE'O2 after 2 min was significantly higher than using the face mask (P<0.01) or the modified NasOral system (P<0.01). The time to maximal FE'O2 was significantly shorter using the modified NasOral system than with the face mask or mouthpiece (P<0.001 and P=0.0001). The volunteers gave more positive ratings to the face mask and mouthpiece than to the modified NasOral system (P<0.001 and P<0.01). We conclude that the use of a mouthpiece can improve preoxygenation in some patients. The results obtained with the modified NasOral system do not justify its introduction into clinical practice. PMID- 11878699 TI - Epidural haematoma after dural puncture in a parturient with neurofibromatosis. AB - A case of epidural analgesia in a parturient with neurofibromatosis (von Recklinghausen's disease) complicated by dural puncture and epidural haematoma is described and the management of the case is discussed. The case emphasizes the need for antenatal assessment of parturients with neurofibromatosis in order that the necessary investigations can be arranged and informed consent for analgesia and anaesthesia can be obtained. PMID- 11878700 TI - Epidural analgesia in a child with sickle cell disease complicated by acute abdominal pain and priapism. AB - We describe a case of a 9-yr-old child with sickle cell disease complicated by abdominal vaso-occlusive crisis and priapism. Both complications were successfully treated with a combination of epidural local anesthetics and morphine. PMID- 11878701 TI - Successful defibrillation in the prone position. AB - Early defibrillation provides the greatest chance of survival after ventricular fibrillation. Conventional cardiopulmonary resuscitation and defibrillation requires the patient to be in the supine position. Electrical treatment of arrhythmias such as atrial fibrillation by means of a defibrillator back paddle in patients receiving prone ventilation in intensive care has been described. We report a case in which electrical defibrillation was successfully performed in the prone position in a patient undergoing complex spinal surgery. We suggest that, if defibrillation were required in ventilated patients positioned prone, defibrillation should be attempted in the prone position, as turning the patient supine would consume valuable minutes and reduce the chances of successful defibrillation. PMID- 11878702 TI - Airway obstruction and microsleep after surgery. PMID- 11878703 TI - Anaphylactic reactions due to chlorhexidine allergy. PMID- 11878704 TI - Autologous transfusion in total knee replacement surgery. PMID- 11878705 TI - Numbness a greater problem than pain? PMID- 11878706 TI - Ondansetron after Caesarian section. PMID- 11878707 TI - Asystole during anaesthetic induction with remifentanil and sevoflurane. PMID- 11878708 TI - Grass allergen-specific T-cells of atopic patients. AB - The prevalence of atopic diseases has increased dramatically in the Western world over the last 20 years. Further insight into the mechanisms of the allergic reaction may help in the development of new more efficient protocols for the specific treatment of allergic diseases. Until a decade ago the major focus in allergy research was on the IgE reactivity of atopic patients and the description of major allergens responsible for the allergic reactions. More recently, the important role of CD4+ T-cells in orchestrating the production of IgE and the maturation of eosinophils through the production of IL-4 (or IL-13) and IL-5 has been clearly established. This has led to the characterization of T-cell responsiveness to numerous major allergens, demonstrating the importance of T cells for the allergic response of atopic patients and for the specific treatment of allergic diseases. One of the major objectives of allergy research is the development of improved treatment protocols for specific therapy. In this review our results on the T-cell reactivity of grass allergic patients to the group 5 major allergen of the grass Phleum pratense (Phl p 5) are discussed in relation to the literature on allergen-specific T-cell reactivity and the specific treatment of allergic disease. Our investigations focused on three subjects: 1) Grass allergen-specific T-cell reactivity of atopic patients in freshly isolated PBMC cultures, 2) Phl p 5 isoallergen recognition of T-cell lines (TCL) and clones (TCC) established from PBMC of grass allergic patients, and 3) Identification of T-cell epitopes on Phl p 5. PMID- 11878709 TI - An association between chronic infection with Chlamydia pneumoniae and lung cancer. A prospective 2-year study. AB - This study assesses a possible relationship between chronic Chlamydia pneumoniae (Cpn) infection and lung cancer (LC). A total of 210 consecutive patients (136 M, 74 F) were diagnosed with LC during a 2-year period. Blood was obtained from 128 M and 70 F patients for Cpn serology. Repeat blood specimens were taken after 3 months. Throat specimens for Cpn DNA analysis by PCR were taken from 110/136 M and 63/74 F Seventy-four cytobrush specimens were taken and also analyzed by polymerase chain reaction (PCR). Fifty (29 M, 21 F) bronchial biopsies and 8 (6 M, 2 F) tumors resected at surgery were analyzed for Cpn by immunohistochemistry (IHC). Males had significantly more often squamous-cell carcinoma (SCC) than females. Other types of LC were more equally distributed between males and females. The difference between males and females regarding smoking history was significant, and male LC patients had significantly higher levels of IgG and/or IgA antibodies than female LC patients. Male and female LC patients had significantly higher prevalences of high antibody titers than controls. A high prevalence of unusually high titers of specific Cpn antibodies was found in male LC patients. This could indicate that LC may be induced by chronic Cpn infection, since stable high titers of Cpn antibodies, especially IgA, are a hallmark of chronic infections. PMID- 11878711 TI - Validity of wet-mount bacterial morphotype identification of vaginal fluid by phase-contrast microscopy for diagnosis of bacterial vaginosis in family practice. AB - The aim of this study was to determine the accuracy of a wet-mount bacterial morphology scoring (BMS) system and Nugent's Gram stain analysis for the diagnosis of bacterial vaginosis, using Amsel's criteria as the gold standard. The three diagnostic criteria were assessed independently The BMS diagnosis was based on a scoring system which weighed the number of small bacterial morphotypes regarded as typical of bacterial vaginosis against lactobacillary morphotypes in phase-contrast microscopy of wet-mount preparations. Three groups of non-pregnant women attending either because of vaginal discharge, other genitourinary symptoms, or for a routine check-up, and a group of pregnant women attending for antenatal care were studied. The diagnostic accuracy was measured by sensitivity, specificity, positive and negative predictive values, and likelihood ratio. The accuracy of the BMS diagnosis was substantially high in all of the examined groups (LR 15.4-20.3). The accuracy of the Gram stain diagnosis was lower (LR 7.6 10.9). In the total material, the accuracy of the BMS diagnosis was higher than that of the Nugent's Gram staining. Intra- and inter-observer reproducibility of all three criteria applied was high. We propose greater routine use of the new BMS diagnosis for point-of-care testing in family practice as well as in research and in microbiology laboratories. PMID- 11878710 TI - Early dissemination of Borrelia burgdorferi without generalized symptoms in patients with erythema migrans. AB - The diagnosis of erythema migrans (EM) is not always easy, and reports of culture or PCR-confirmed diagnosis as well as reports of EM with simultaneous disseminated disease are few. Characteristics and incidence of EM in addition to frequency of early dissemination of B. burgdorferi were studied in the archipelago of South-Western Finland prospectively using questionnaires, skin biopsies and blood samples. Clinical EM was recognized in 82 patients (incidence 148/100,000 inhabitants/year). Of skin biopsy samples, 35.5% were positive by PCR (the majority B. garinii), and 21.5% by cultivation (all B. garinii). Of blood samples, 3.8% were positive by PCR, and 7.7% by cultivation. Of the patients, 30.9% were seropositive at the first visit, and 52.9% 3 weeks later. Of the patients with laboratory confirmed diagnosis, the EM lesion was ring-like in 31.8% and homogeneous in 65.9%. Dissemination of B. burgdorferi, based on culture or PCR positivity of blood samples, was detected in 11.0% of the patients. The frequency of generalized symptoms was nearly the same in patients with as in those without dissemination (22.2% vs 27.4%). Only 21.4% of the patients with culture-positive EM recalled a previous tick bite at the site of the EM lesion. We conclude that EM lesions are more often homogeneous than ring-like. B. burgdorferi may disseminate early without generalized symptoms. PMID- 11878712 TI - Urethritis associated with Chlamydia trachomatis: comparison of leukocyte esterase dipstick test of first-voided urine and methylene blue-stained urethral smear as predictors of chlamydial infection. AB - The use of nucleic acid amplification tests for the diagnosis of C. trachomatis has made it possible to send urine samples instead of urethral swab specimens to the laboratory. The sensitivity is very high, but not 100%, and we continue to perform a test for urethritis at our STD clinic. The aim of this study was to compare the performance of two alternative tests in the diagnosis of urethritis as predictors of C. trachomatis infection: the leukocyte esterase (LE) dipstick test of first-voided urine and polymorphonuclear leukocyte counts in a methylene blue-stained (MBS) urethral smear. Urine samples from 480 male patients attending an STD clinic were analysed using the LE test and LCR assay for C. trachomatis; urethral samples were analysed with MBS urethral smear and LCR. The majority (75.8%) of the 480 patients examined were asymptomatic. Chlamydial infection was detected in 50 patients. The sensitivity, specificity and positive predictive value of the LE test for predicting C. trachomatis infection were 46.0, 91.6 and 39.0%, respectively, among all patients examined and 25.9, 95.8 and 33.3%, respectively, among the asymptomatic patients. The corresponding values for the MBS urethral smear were 76.0, 82.1 and 33.0% among all patients and 63.0, 89.6 and 32.7% among the asymptomatic patients. At our STD clinic we chose to perform the examination of MBS urethral smears in the diagnosis of urethritis because of its higher sensitivity relative to the LE test for predicting C. trachomatis. PMID- 11878713 TI - Benign solitary fibrous pleural tumour. Evidence of primitive features and complex genomic imbalances, including loss of 20q. AB - AIMS: Cytogenetic data on solitary fibrous tumours (SFT) are very limited. We studied a benign pleural SFT for its ultrastructural and immunohistochemical details, and made cytogenetic analyses for comparison with other genetic and ultrastructural studies of SFT. RESULTS: Immunohistochemistry showed strong positivities for CD34 and vimentin, but no reactions with anti-cytokeratins and epithelial membrane antigens. Electron microscopy revealed primitive desmosomes in our SFT. The results thus evinced fibroblast-like cells with intermediate epithelial-mesenchymal character. Comparative genomic hybridization of the tumour revealed losses of 1p33-->pter, 17pter q21, entire copies of chromosomes 19 and 22, and gains of 1p21-p22, 2q23-q32.3, 3pl2-q13.2, 4p14-q28, 6p12-q21, 9p21- >pter and 13q21-q31. Furthermore, there was loss of 20q, as was previously reported elsewhere in a case of benign and a case of malignant SFT. CONCLUSIONS: The results furnish further evidence of the involvement of -20q in SFT. In addition, they show that SFT may have complex genomic imbalances and primitive features, despite having a benign appearance. PMID- 11878714 TI - Telomerase activity, MIB-1, PCNA, HPV 16 and p53 as diagnostic markers for cervical intraepithelial neoplasia. AB - The present investigation evaluated the relationship between dysplasia of the uterine cervix and telomerase activity, expression of p53, MIB-1 and PCNA. Telomerase activity was measured on cervical cytobrush material from 126 women suspected of having dysplasia and 61 controls using the telomeric repeat amplification protocol. Immunohistochemistry was used to detect the tumor suppressor protein p53 and cell proliferation, the latter by MIB-1 and PCNA expression. Infection with human papillomavirus 16 was detected by PCR amplification and Southern blot hybridization of DNA extracted from the same brush material. Positive telomerase activity was found in 5 of 43 (11.6%) normal samples, 12 of 57 (21.1%) samples with inflammation or koilocytosis, 7 of 17 (41.2%) CIN 1 (cervical intraepithelial neoplasia, grade 1), 8 of 20 (40.0%) CIN 2, and 25 of 42 (59.5%) CIN 3/ CIS. Telomerase activity was significantly related to the level of dysplasia (p<0.001) and proliferation measured by MIB-1 (p=0.019), but not to the level of PCNA (p=0.445), HPV 16 status (p=0.098) or staining for p53 (p=0.271). Dysplasia was also related to PCNA, MIB1, p53, and presence of HPV 16. A sequential increase in the examined parameters, paralleling the progression of abnormality, was observed. PCNA and telomerase showed an increase in CIN 1, MIB-1 and HPV16 in CIN 2, and finally p53 in CIN 3/CIS. PMID- 11878715 TI - Neutrophil free oxygen radical production and blood total antioxidant capacity in patients with coronary heart disease using various medications. AB - Despite convincing results of studies in vitro, less is known about the effects of antioxidants on in vivo redox balance in humans. We developed a novel parameter of in vivo redox balance, and studied it and its relation to dental infections in 51 patients on medication for coronary heart disease (CHD) and 39 random controls matched for age group, sex, social class and locality. In vivo redox balance was the ratio of plasma antioxidant capacity, as measured with radical-trapping assay, to neutrophil respiratory burst capacity, as measured with whole blood chemiluminescence assay. Dental infections were quantitated with four rating scales. CHD patients had higher values than controls. Patients on acetosalicylic acid (ASA), diuretics or beta blockers, but not the ones on calcium channel blocker, had significantly higher redox balance than non-users. Combination of calcium channel blockers and ASA was associated with redox balance similar to taking beta blockers or diuretics. Diuretics and ASA were independent determinants of redox balance in multivariate analyses. Redox balance did not correlate with severity of dental infections (Spearman's r 0.06 to 0.11). The results contrast experimental data indicating that calcium channel blockers are as antioxidants superior to other cardiovascular drugs. Total antioxidant capacity in parallel with oxygen species production capacity should be considered in attempts to solve the antioxidant paradox. PMID- 11878716 TI - Abnormal venous structures in salivary gland tumors: vasculature characteristics of pleomorphic adenoma. AB - To clarify the diagnostic significance of abnormal venous structures present in salivary gland tumors, we examined 21 pleomorphic adenomas, 14 Warthin tumors, 1 oncocytic adenoma, 3 myoepitheliomas, 7 basal cell adenomas, 5 mucoepidermoid carcinomas, and 6 adenoid cystic carcinomas. Verhoeffvan Gieson staining was carried out and the morphology of the veins within the tumors was observed microscopically. Branching veins, thickened intima of the veins, discontinuous elastic membrane and multilayered elastic membrane were seen in 71.4%, 76.2%, 47.6% and 85.7% of pleomorphic adenomas, respectively, and were abundant and easily found in most cases. The abnormal venous structures were also found in other salivary gland tumors examined, but they were few in number and lacked variety. Elastic fibers extending radially into the surrounding stroma were seen in 66.7% of pleomorphic adenomas, and were not seen in other salivary gland tumors. Our results showed that a variety of abnormal venous structures are more abundant and more easily found in pleomorphic adenoma compared with other salivary gland tumors, and, in particular, that perivascular radiating elastic fibers are characteristic of pleomorphic adenoma. We emphasize that the presence of perivascular radiating elastic fibers may be helpful in diagnosing pleomorphic adenoma in small biopsy specimens. PMID- 11878717 TI - Atypical decubital fibroplasia with unusual histology. AB - A case of atypical decubital fibroplasia with unusual histology arising in the buttock of a 68-year-old bed-ridden male in presented. The lesion measuring 5.4 cm in greatest dimension was histologically characterized by a proliferation of fibroblasts with oval to spindle nuclei and dense fibrous stroma with focal hyalinization and calcification. Ganglion-like fibroblastic cells and multinucleated giant cells of osteoclast type were also observed. There were numerous elastic fibers within and adjacent to the proliferating stromal cells. The proliferating stromal cells were positive for vimentin and collagen type IV but negative for CAM 5.2, epithelial membrane antigen, desmin, alpha-smooth muscle actin, muscle actin, HHF35, S-100 protein and CD34. Ultrastructurally, they were of a fibroblastic nature. The hypercellularity, lack of zones of fibrinoid necrosis, lack of lobulation and the presence of multinucleated giant cells were different from the originally described lesion. This condition represents a variant of atypical decubital fibroplasia. Pathogenic factors of this lesion are considered to be chronically repeated pressure and associated intermittent ischemia. The recognition of the lesion and its distinction from a sarcoma is essential. PMID- 11878718 TI - Chlamydia trachomatis infection in mothers with preterm delivery and in their newborn infants. AB - We studied Chlamydia trachomatis infection in mothers with preterm delivery and intrauterine transmission of the infection to their offspring. Forty-one mothers with preterm labour and their newborn infants (n=50) were studied for the presence of C. trachomatis infection using microimmunofluorescence test for detection of serum antibodies against C trachomatis and polymerase chain reaction for detection of C. trachomatis-specific DNA in mucosal swabs. Antibodies to C trachomatis were found in serum of 12 mothers (29%). Five of fourteen mothers had C. trachomatis DNA in cervical specimens. Eighteen neonates were born to the 14 mothers with positive serology and/or C. trachomatis DNA. C. trachomatis DNA was detected in specimens from 10 of the 18 neonates (55.5%). Three of the available cord blood samples contained C trachomatis IgM antibodies. Our results strongly suggest that mothers and their preterm babies may benefit from screening for active C. trachomatis infection. PMID- 11878719 TI - Measuring the outcome of surgical procedures: what are the challenges? PMID- 11878720 TI - Time to light the grey touchpaper! The challenge of anaesthesia in the elderly. PMID- 11878721 TI - Relationship of the functional recovery after hip arthroplasty to the neuroendocrine and inflammatory responses. AB - We studied the relationship between the neuroendocrine and inflammatory responses to hip arthroplasty and functional recovery in 102 patients undergoing elective arthroplasty for osteoarthritis. Blood samples were collected for up to 7 days after surgery and analysed for concentrations of norepinephrine, epinephrine, cortisol, interleukin-6 and C-reactive protein. The primary outcome measures were milestones in hospital, times to walk 10 and 25 m, pain on discharge from hospital, and function 1 and 6 months after surgery. Walking distances in hospital were significantly delayed in patients with greater interleukin 6 and C reactive protein concentrations, but few neuroendocrine measures had significant correlations with functional recovery in hospital. Multivariate analysis showed that the interleukin 6 concentration on day 1 was the unique predictor of time to walk 10 and 25 m, and that the day 2 concentration of C-reactive protein was the unique predictor of pain on discharge from hospital. No significant correlations were found between the inflammatory and neuroendocrine variables and recovery at 1 and 6 months. We conclude that the inflammatory response affects immediate functional recovery after hip arthroplasty. PMID- 11878722 TI - Development and testing of a hierarchical method to code the reason for admission to intensive care units: the ICNARC Coding Method. Intensive Care National Audit & Research Centre. AB - A computer-based hierarchical method was developed to code conditions leading to admission to intensive care in the UK. The hierarchy had five tiers: surgical status, body system, anatomical site, physiological or pathological process and medical condition. The hierarchy was populated initially using the free-text descriptions of the reason for admission from 10,806 admissions recorded as part of the Intensive Care Society's UK APACHE II study. After refinement and error checking, a prospective evaluation was undertaken on 22,059 admissions to 62 UK intensive care units. Individual units coded between 60 and 1610 (mean 356) admissions. All but 50 (0.2%) of the admissions could be coded and 38 units coded every admission. Fifty admissions (0.2%) could not be coded within 24 h of admission but were coded subsequently when more information became available. Of the admissions, 96.1% were coded at all levels of the hierarchy in the coding method. Six hundred and thirty-seven of the 741 unique conditions (85.9%) were used in one of the five reasons for admission and 564 (76.1%) in the primary reason for admission. Five conditions account for 19.4% of all primary reasons for admission. This is the first method to be developed empirically for coding the reason for intensive care admission. PMID- 11878723 TI - Anaphylaxis during anaesthesia. Results of a two-year survey in France. AB - Between January 1, 1997 and December 31, 1998, 467 patients were referred to one of the allergo-anaesthesia centres of the French GERAP (Groupe d'Etudes des Reactions Anaphylactoides Peranesthesiques) network and were diagnosed as having anaphylaxis during anaesthesia. Diagnosis was established on the basis of clinical history, skin tests and/or a specific IgE assay. The most frequent cause of anaphylaxis was a neuromuscular blocking agent (69.2%). Latex was less frequently incriminated (12.1%) than in previous reports. A significant difference was observed between the incidence of anaphylactic reactions observed with each neuromuscular blocking agent and the number of patients who received each drug during anaesthesia in France throughout the study period (P<0.0001). Succinylcholine and rocuronium were most frequently incriminated. Clinical reactions to neuromuscular blocking drugs were more severe than to latex. The diagnostic value of specific IgE assays was confirmed. These results are consistent with changes in the epidemiology of anaphylaxis related to anaesthesia and are an incentive for the further development of allergo-anaesthesia clinical networks. PMID- 11878724 TI - Measuring the costs of inhaled anaesthetics. AB - The cost of inhalation anaesthesia has received considerable study and is undoubtedly reduced by the use of low fresh gas flows. However, comparison between anaesthetics of the economies achievable has only been made by computer modelling. We have computed anaesthetic usage for MAC-equivalent anaesthesia with isoflurane, desflurane, and sevoflurane in closed and open breathing systems. We have compared these data with those derived during clinical anaesthesia administered using a computer-controlled closed system that measures anaesthetic usage and inspired concentrations. The inspired concentrations allow the usage that would have occurred in an open system to be calculated. Our computed predictions lie within the 95% confidence intervals of the measured data. Using prices current in our institution, sevoflurane and desflurane would cost approximately twice as much as isoflurane in open systems but only about 50% more than isoflurane in closed systems. Thus computer predictions have been validated by patient measurements and the cost saving achieved by reducing the fresh gas flow is greater with less soluble anaesthetics. PMID- 11878725 TI - Improvement of 'dynamic analgesia' does not decrease atelectasis after thoracotomy. AB - There is still controversy concerning the beneficial aspects of 'dynamic analgesia' (i.e. pain while coughing or moving) on the reduction of postoperative atelectasis. In this study, we tested the hypothesis that thoracic epidural analgesia (TEA) prevents these abnormalities as opposed to multimodal analgesia with i.v. patient controlled analgesia (i.v. PCA) after thoracotomy. Fifty-four patients undergoing thoracotomy (lung cancer) were randomly assigned to one of the two groups. Clinical respiratory characteristics, arterial blood gas, and pulmonary function tests (forced vital capacity and forced expiratory volume in 1 s) were obtained before surgery and on the next 3 postoperative days. Atelectasis was compared between the two groups by performing computed tomography (CT) scan of the chest at day 3. Postoperative respiratory function and arterial blood gas values were reduced compared with preoperative values (mean (SD) FEV1 day 0: 1.1 (0.3) litre; 1.3 (0.4) litre) but there was no significant difference between groups at any time. PCA and TEA provided a good level of analgesia at rest (VAS day 0: 21 (15/100); 8 (9/100)), but TEA was more effective for analgesia during mobilization (VAS day 0: 52 (3/100); 25 (17/100)). CT scans revealed comparable amounts of atelectasis (expressed as a percentage of total lung volume) in the TEA (7.1 (2.8)%) and in the i.v. PCA group (6.71 (3.2)%). There was no statistical difference in the number of patients presenting with at least one atelectasis of various types (lamellar, plate, segmental, lobar). PMID- 11878726 TI - Efficacy and uptake of ropivacaine and bupivacaine after single intra-articular injection in the knee joint. AB - The efficacy of ropivacaine 100 mg (5 mg ml(-1)), 150 mg (7.5 mg ml(-1)) and 200 mg (10 mg ml(-1)) and bupivacaine 100 mg (5 mg ml(-1)) given by intra-articular injection into the knee after the end of surgery was studied in 72 ASA I-II patients scheduled for elective knee arthroscopy under general anaesthesia in a randomized, double-blind study. Kapake (paracetamol 1 g and codeine 60 mg) was given as a supplementary analgesic. Pain scores were assessed 1-4 h after surgery and a verbal rating scale of overall pain severity was assessed on second postoperative day. Ropivacaine or bupivacaine concentrations were determined in peripheral venous plasma up to 3 h after injection in eight patients in each group. Verbal rating pain scores were lower with ropivacaine 150 mg compared with bupivacaine 100 mg (P<0.05). There was a tendency for lower analgesic consumption and pain scores with all doses of ropivacaine (not significant). The mean (SD) maximum total plasma concentrations of ropivacaine were 0.64 (0.25), 0.78 (0.43), and 1.29 (0.46) mg litre(-1) after 100, 150 and 200 mg. The corresponding unbound concentrations were 0.018 (0.009), 0.024 (0.020) and 0.047 (0.022) mg litre(-1). Both were proportional to the dose. The maximum total concentration after bupivacaine 100 mg was 0.57 (0.36) mg litre(-1). The time to reach maximum plasma concentration was similar for all doses and varied between 20 and 180 min. All concentrations were well below the threshold for systemic toxicity. PMID- 11878727 TI - Effect of peri- and postoperative epidural anaesthesia on pain and gastrointestinal function after abdominal hysterectomy. AB - In a double blind study we have investigated the effects of epidural local anaesthesia (LA), when added to general anaesthesia (GA) and postoperative paracetamol and NSAID, on postoperative pain and gastrointestinal function in patients undergoing open hysterectomy. Sixty patients were randomized into three study groups: GA, and postoperative paracetamol and NSAID (GA, n=20); GA, paracetamol, NSAID, intraoperative epidural lidocaine and 24-h postoperative epidural saline (Saline, n=20); or GA, paracetamol, NSAID, intraoperative epidural lidocaine and 24-h postoperative epidural bupivacaine (Bupi, n=20). Patients were observed for 72 h postoperatively. Pain at rest, during cough, and mobilization, request for supplementary morphine, and time to first postoperative flatus, was reduced in patients receiving 24-h postoperative epidural anaesthesia, compared with the two other groups. However, these effects of epidural LA, were not sustained beyond the period of infusion, and no differences in PONV, time to first postoperative defecation, mobilization or time to discharge from hospital were observed between groups. A 24 h postoperative epidural infusion with bupivacaine, when added to postoperative paracetamol and NSAID, reduces pain and opioid requirements, but has only limited effects on gastrointestinal function and patient recovery. PMID- 11878728 TI - Comparison of articaine and bupivacaine/lidocaine for single medial canthus peribulbar anaesthesia. AB - In a single-centre, randomized, double-blind study, we compared the efficacy of 2% articaine with that of a mixture of 0.5% bupivacaine and 2% lidocaine for peribulbar anaesthesia in cataract surgery, using a single medial canthus injection technique. Eighty-two patients were allocated randomly to receive 7-9 ml of a mixture of 0.5% bupivacaine and 2% lidocaine or an equal volume of 2% articaine with 1:200,000 epinephrine. Hyaluronidase 30 iu ml(-1) was added to both solutions. The degree of akinesia was scored 1, 5 and 10 min after the block, at the end of surgery and at discharge from the day case unit. Primary outcome measures were the difference in ocular movement scores 5 min after block and the need for supplementary inferolateral injections. There was greater akinesia in the articaine group at 5 min (P=0.01). Ten patients (24%) in the articaine group and 21 patients (51%) in the bupivacaine/lidocaine group required a supplementary injection (P=0.02). The mean (SD) volume of local anaesthetic required to achieve adequate block for surgery was 9.7 (2.1) ml in the articaine group and 11.0 (2.2) ml in the bupivacaine/lidocaine group (P=0.01). There was a faster offset of akinesia after surgery in the articaine group (P=0.01). There were no differences between groups in the incidence of reported pain or of minor complications. In our study, 2% articaine with 1:200,000 epinephrine was safe and efficacious for single medial canthus peribulbar anaesthesia. PMID- 11878729 TI - Prospective randomized, double-blind comparative study of dexamethasone, ondansetron, and ondansetron plus dexamethasone as prophylactic antiemetic therapy in patients undergoing day-case gynaecological surgery. AB - Dexamethasone alone and in combination with selective 5-hydroxytryptamine receptor antagonists is of benefit in the prophylaxis of post-operative nausea and vomiting. In this study, the effectiveness of such a combination in comparison to either drug alone is investigated in day case gynaecological surgery. A total of 177 patients were randomized to three treatment groups: dexamethasone 8 mg, ondansetron 4 mg, and dexamethasone 8 mg plus ondansetron 4 mg. The only significant difference between groups was seen in the first 3 h when failure of prophylaxis was more frequent in patients who had received dexamethasone alone (P=0.0085; Fisher's exact probability test). Confidence interval analysis indicates a modest treatment effect for the combination and the decision whether to perform a larger study depends upon whether such an effect is clinically relevant. PMID- 11878730 TI - Small dose of exogenous surfactant combined with partial liquid ventilation in experimental acute lung injury: effects on gas exchange, haemodynamics, lung mechanics, and lung pathology. AB - A combination of exogenous surfactant and partial liquid ventilation (PLV) with perfluorocarbons should enhance gas exchange, improve respiratory mechanics and reduce tissue damage of the lung in acute lung injury (ALI). We used a small dose of exogenous surfactant with and without PLV in an experimental model of ALI and studied the effects on gas exchange, haemodynamics, lung mechanics, and lung pathology. ALI was induced by repeated lavages (PaO2/FIO2 less than 13 kPa) in 24 anaesthesized, tracheotomized and mechanically ventilated (FIO2 1.0) juvenile pigs. They were treated randomly with either a single intratracheal dose of surfactant (50 mg kg(-1), Curosurf, Serono AG, Munchen, Germany) (SURF-group, n=8), a single intratracheal dose of surfactant (50 mg kg(-1), Curosurf) followed by PLV with 30 ml kg(-1) of perfluorocarbon (PF 5080, 3M, Germany) (SURF-PLV group, n=8) or no further intervention (controls, n=8). Pulmonary gas exchange, respiratory mechanics, and haemodynamics were measured hourly for a 6 h period. In the SURF-group, the intrapulmonary right-to-left shunt (QS/QT) decreased significantly from mean 51 (SEM 5)% after lavage to 12 (2)%, and PaO2 increased significantly from 8.1 (0.7) to 61.2 (4.7) kPa compared with controls and compared with the SURF-PLV-group (P<0.05). In the SURF-PLV-group, QS/QT decreased significantly from 54 (3)% after induction of ALI to 26 (3)% and PaO2 increased significantly from 7.2 (0.5) to 30.8 (5.0) kPa compared with controls (P<0.05). Static compliance of the respiratory system (C(RS)), significantly improved in the SURF-PLV-group compared with controls (P<0.05). Upon histological examination, the SURF-group revealed the lowest total injury score compared with controls and the SURF-PLV-group (P<0.05). We conclude that in this experimental model of ALI, treatment with a small dose of exogenous surfactant improves pulmonary gas exchange and reduces the lung injury more effectively than the combined treatment of a small dose of exogenous surfactant and PLV. PMID- 11878731 TI - Viscosity and density of common anaesthetic gases: implications for flow measurements. AB - Although viscosity (mu) is a crucial factor in measurements of flow with a pneumotachograph, and density (rho) also plays a role in the presence of turbulent flow, these material constants are not available for the volatile anaesthetic agents commonly administered in clinical practice. Thus, we determined experimentally mu and rho of pure volatile anaesthetic agents. Input impedance of a rigid-wall polyethylene tube (Zt) was measured when the tube was filled with various mixtures of carrier gases (air, 100% oxygen, 50% oxygen+50% nitrogen) to which different concentrations of volatile anaesthetic inhalation agents (halothane, isoflurane, sevoflurane, and desflurane) had been added. Mu and rho were calculated from real and imaginary portions of Zt, respectively, using the appropriate physical equations. Multiple linear regression was applied to estimate mu and rho of pure volatile agents. Viscosity values of pure volatile agents were markedly lower than those for oxygen or nitrogen. Clinically applied concentrations, however, did not markedly affect the viscosity of the gas mixture (maximum of 3.5% decrease in mu for 2 MAC desflurane). In contrast, all of the volatile agents significantly affected rho even at routinely used concentrations. Our results suggest that the composition of the carrier gas has a greater impact on viscosity than the amount and nature of the volatile anaesthetic agent whereas density is more influenced by volatile agent concentrations. Thus, the need for a correction factor in flow measurements with a pneumotachograph depends far more on the carrier gas than the concentration of volatile agent administered, although the latter may play a role in particular experimental or clinical settings. PMID- 11878732 TI - Minimizing perioperative adverse events in the elderly. AB - Elderly patients still have the highest postoperative mortality and morbidity rate in the adult surgical population. Preoperative clinical assessment to detect patients at high risk of postoperative events, and specific intraoperative and postoperative anaesthesia management are important to minimize postoperative adverse events in the elderly. PMID- 11878733 TI - Neuromuscular monitoring in intensive care patients: milliamperage requirements for supramaximal stimulation. AB - We investigated the effects of peripheral oedema on the supramaximal current required for neuromuscular monitoring of critically ill patients. We studied 32 sedated patients who had not needed a neuromuscular blocking drug. The presence of oedema over the volar aspect of both wrists was assessed by a blinded observer and graded (grade 0, no oedema; grade 1, mild oedema; grade 2, gross oedema). The supramaximal current was derived by applying an incrementally increasing current over the ulnar nerve and measuring the amplitude of the electromyographic (EMG) response of the first dorsal interosseous muscle. The supramaximal current was that current above which there was no significant increase in EMG amplitude. It was 40 mA in the absence of oedema. This current was significantly increased in the presence of grade 1 oedema (60 mA, Mann-Whitney test, P<0.01) and grade 2 oedema (82.5 mA, Mann-Whitney test, P<0.01). In the presence of oedema, the required supramaximal current decreased significantly after the application of pressure over the stimulating electrodes (Wilcoxon signed rank test, P<0.05). Supramaximal current in critically ill patients is increased in the presence of peripheral oedema. We recommend that nerve stimulators used for neuromuscular monitoring in the ICU are capable of delivering a stimulus current of at least 100 mA. PMID- 11878734 TI - Contamination of coagulation tests with heparin from blood gas samples. AB - We investigated the possibility that samples of blood could be contaminated by heparinized blood gas syringes, giving spurious results in coagulation tests. We collected coagulation test samples before (n=5) and after (n=13) blood gas sampling from a venous cannula in a volunteer. The results of activated partial thromboplastin time (APTT) tests were compared between control samples and samples taken before and after blood gas sampling. The median APTT after blood gas sampling was 71 s, significantly higher than before (median 34 s, P<0.05). We conclude that heparin from blood gas sampling syringes can contaminate coagulation tests. Heparinized samples for blood gas analysis should be obtained after coagulation test samples have been obtained. PMID- 11878735 TI - Randomized prospective double-blind placebo-controlled trial of effect of intravenous ondansetron on intraocular pressure during ophthalmic surgery. AB - The effect of i.v. ondansetron, before induction of anaesthesia, on intraocular pressure (IOP) in patients undergoing cataract surgery was investigated. Forty patients (two groups of 20) received either ondansetron 4 mg (treatment group) or 0.9% saline (placebo group) in a double-blind controlled manner. There were no significant differences in IOP between the groups. Ondansetron had no significant effect on IOP during the study period. PMID- 11878736 TI - Evaluation of the Greenbaum sub-Tenon's block. AB - A prospective, randomized blind study was conducted in 40 patients undergoing phacoemulsification and posterior chamber intraocular lens implantation. They received anaesthetic infiltration of 2% lidocaine with 1:200,000 epinephrine and hyaluronidase 150 U ml(-1) in a volume of 2, 3, 4 or 5 ml into the sub-Tenon's fascial space through a Greenbaum cannula after a conjunctival incision. Reduction of ocular movements, anaesthesia, pain on injection and any incidental complications were recorded. Akinesia and anaesthesia occurred within 5 min with 4 and 5 ml of local anaesthetic, and no supplementary injections were required. There were marked reductions in the frequency of forced eyelid movements with these volumes. Chemosis and conjunctival haemorrhage were noted in the majority of patients but caused no intraoperative problems. Approximately 10-15% of patients reported slight discomfort at the time of injection. Four to 5 ml of 2% lidocaine with 1:200,000 epinephrine and 150 U ml(-1) of hyaluronidase is the optimum volume to achieve adequate akinesia, anaesthesia and reduction of lid movements during the Greenbaum sub-Tenon's block. PMID- 11878737 TI - Comparison of patient-controlled epidural analgesia with and without night-time infusion following gastrectomy. AB - To assess the analgesic efficacy and side effects of a supplemental night-time infusion in patient-controlled epidural analgesia (PCEA) after gastrectomy, we carried out a randomized, double-blind study. The number of requests were lower (P<0.005) in the PCEA plus night-time infusion group than in the PCEA alone group during the postoperative nights. Patients who had a PCEA plus night-time continuous infusion, slept with fewer interruptions than those who had only the PCEA. VAS pain scores on coughing were significantly lower (P<0.05) in the PCEA plus infusion group than in the PCEA alone group during the night following postoperative day 1. In conclusion, a night-time infusion in PCEA following gastrectomy decreases the incidence of postoperative pain, provides a better sleep pattern, and reduces the degree of the pain associated with coughing during the night. PMID- 11878738 TI - Postoperative analgesia by epidural methylprednisolone after posterolateral thoracotomy. AB - The aim of this study was to evaluate the potential analgesic effect of epidural methylprednisolone (MP) after posterolateral thoracotomy (PLT). Adult male patients undergoing PLT for lung surgery were included in a prospective, randomized, double blind study. Peroperative analgesia (bupivacaine plus sufentanil) was given by a thoracic epidural catheter associated with general anaesthesia. After surgery, patients received either MP 1 mg kg(-1) followed by a continuous epidural infusion of MP 1.5 mg kg(-1) during 48 h (MP group) or 0.9% saline as a bolus injection and continuous epidural infusion (P group). Additional morphine analgesia was administered by i.v. patient-controlled analgesia. Pain was assessed at rest and with mobilization every 4 h after operation during 48 h with a visual analogue scale (VAS). The primary end-point was the total morphine requirements during the 48 first postoperative hour. Twenty-four patients were allocated to MP (n=12) and P (n=12) groups. Characteristics of the two groups were similar. There were no differences between groups for morphine requirements (median and interquartile range) during the 48 h: 59 mg (40-78) in MP group vs 65 mg (59-93) in P group. There were no differences between groups for morphine requirements every 4 h during the 48 h and VAS for pain at rest and evoked pain. No side effects were reported. It was concluded in this small study that these results did not support the use of epidural steroids for postoperative analgesia after PLT. PMID- 11878739 TI - Serum paracetamol concentrations in adult volunteers following rectal administration. AB - Paracetamol is usually given in adults at a dose of 10-20 mg kg(-1) orally or rectally. Work in children suggests that doses of 40 mg kg(-1) are needed to provide therapeutic concentrations when this drug is used by the rectal route. We have investigated the dose of rectal paracetamol needed to achieve serum concentrations within the accepted therapeutic range of 10-20 microg ml(-1) in adults. Ten healthy adult volunteers received increasing doses of rectal paracetamol (15, 25, 35, and 45 mg kg(-1)). Following suppository administration, serum paracetamol concentrations were measured half hourly to 4 h then hourly to 8 h. Sustained concentrations within our therapeutic range were achieved with 35 and 45 mg kg(-1). Maximum measured concentrations were 12.5 (10-16), 16.5 (14 20), and 20 (17.5-23) microg ml(-1), median (inter-quartile range) after 25, 35, and 45 mg kg(-1), respectively. We conclude that doses of 35-45 mg kg(-1) of rectal paracetamol are needed to achieve sustained therapeutic plasma concentrations in healthy adult volunteers. PMID- 11878740 TI - Blood pressure manipulation during awake carotid surgery to reverse neurological deficit after carotid cross-clamping. AB - We describe the management of three patients undergoing awake carotid surgery who developed signs of cerebral ischaemia after carotid cross-clamping. Drug treatment to increase arterial blood pressure above baseline reversed the neurological deficit and an internal carotid artery shunt was not needed. Shunt insertion is less frequent with regional rather than general anaesthesia, and blood pressure control can reduce this even more. Coincidentally, one of the patients, who gave a history of angina of effort after walking 100 m, complained of chest pain after cross-clamp release. This was treated successfully with sublingual nitroglycerin before ST segment changes became apparent on the ECG. These reports suggest that regional anaesthesia for carotid surgery allows potential complications to be identified earlier than under general anaesthesia using reports from the patient, so that treatment may be modified to prevent morbidity and even mortality. PMID- 11878741 TI - Cardiac arrest associated with use of an argon beam coagulator during laparoscopic cholecystectomy. AB - We describe a cardiac arrest during use of an argon beam coagulation (ABC) system in an 82-yr-old woman having laparoscopic cholecystectomy under general and epidural anaesthesia. Intra-abdominal pressure (IAP) was controlled to less than 12 mm Hg during a carbon dioxide gas pneumoperitoneum and at first the operation was uneventful. When the ABC system (gas flow 6 litre min(-1)) was used to control local bleeding in the liver bed abdominal pressure increased rapidly to over 20 mm Hg and, 1 min later, the end-tidal carbon dioxide decreased to zero, followed by bradycardia and cardiac arrest. At once, an emergency laparotomy was performed and resuscitation begun. A mill-wheel murmur was heard on auscultation, leading to suspicion of argon gas embolism. Fortunately, recovery was completed with no neurological deficit. Anaesthesiologists should consider showed that argon gas embolism can occur with the ABC system during laparoscopic surgery. PMID- 11878742 TI - Assessment instruments used during anaesthetic simulation. PMID- 11878743 TI - Renal dysfunction following anaesthesia and surgery. PMID- 11878744 TI - Perioperative bradycardia. PMID- 11878745 TI - Management of purpura fulminans. PMID- 11878746 TI - Undiagnosed myasthenia gravis unmasked by neuromuscular blockade. PMID- 11878747 TI - Selenium status and its correlates in a British national diet and nutrition survey: people aged 65 years and over. AB - Plasma selenium (Se) concentrations were measured in 1134 British people aged 65 years and over, living in mainland Britain during 1994-1995. Eight hundred and eighty-three lived in the community ("free-living"), while the remainder lived in institutions (residential and nursing homes). The overall mean plasma Se concentrations was 0.90 micromol/l (inner 95% range 0.50-1.36 micromol/l). Free living people had significantly higher values than counterparts living in institutions, and there was an overall inverse correlation with age. Plasma Se varied with season (lower from October to December than at other times of the year), while values were higher in southern Britain than in the North. Socio economic associations also existed (lower plasma Se in people receiving state benefits or with poorer educational attainment), while smokers had lower values than non-smokers. Several indices of poor health status or of medicine use were correlated with lower plasma Se, which was also predicted by several biochemical or haematological indices of infection or inflammation. Plasma Se was strongly and directly correlated with plasma albumin, zinc, cholesterol, vitamin C, several carotenoids, alpha-tocopherol, pyridoxal phosphate, and with blood haemoglobin and several anthropometric indices. All relationships were independent of age and gender, and are consistent with the view that plasma Se levels are higher in fit and well-nourished elderly people and lower in those who are frail, poorly-nourished and unwell. Whole-blood glutathione peroxidase generally did not share these relationships, was only very weakly correlated with plasma Se, and appeared to be less useful as a status indicator in this population group. PMID- 11878748 TI - Multi-element analysis of trace element levels in human autopsy tissues by using inductively coupled atomic emission spectrometry technique (ICP-AES). AB - Autopsy tissue samples from the brain front lobe, cerebellum, heart, kidney (cortex and medulla), liver, pancreas, spleen and ovary were analysed for AL, B, Ba, Cd, Co, Cr, Cu, Fe, Mn, Ni, Pb, Se, Sr and Zn in 30 (17 women and 13 men) subjects ranging in age from 17 to 96 years at Haukeland University Hospital in Norway. The tissues were selected from macroscopically normal organs and samples were handled according to guidelines recommended to avoid contamination in the pre-analytical phase. Concentration of the trace elements were determined by the inductively coupled plasma atomic emission spectrometry technique (ICP-AES). In most tissues the concentrations of the essential trace elements followed the order Fe> Zn> Cu> Mn> Se> Cr> Co except in the ovary where Se was higher than Mn. The liver was the major site of deposition for Co, Cu and Mn as well as the spleen for Co, brain front lobe for Cu and pancreas for Mn. Ba, Sr and Ni built up in the ovary foLLowed by the kidney. Older subjects accumulated Ba and Sr in most tissues, whereas Al accumulated in the kidney cortex and Cd in the brain cerebellum. Generally males had higher concentrations of trace elements in the different tissue sampLes than females with the exception of Mn in the brain front lobe and heart and Sr in the liver. ICP-AES is a useful method to assess the concentration and the profiLe of trace elements in human autopsy tissues. PMID- 11878749 TI - Assessment of hair mineral analysis commercially offered in Germany. AB - To evaluate intra- and inter-laboratory agreement concerning hair mineral analysis and interpretation of results, hair samples from 2 volunteers were sent to seven laboratories, which commercially offer hair mineral analysis in Germany. 6 weeks later, another identical part from the hair sample of volunteer 1 was sent to all seven labs. Altogether, 50 elements were analyzed, 23 by all seven labs. For comparability, only the results for these 23 elements were assessed. The intra-laboratory reproducibility was evaluated by the 2 identical hair samples from volunteer 1. On the average, the reproducibility seems to be sufficient (median +/- 9.48% to +/- 20.59%), but for individual elements there were unacceptable out-rulers up to 100%. Only one lab classified all elements of the first and the second analysis of the identical hair sample in the same category (below, within, or above normal range). The others grouped 4 to 7 elements different. This is not tolerable. The inter-laboratory comparability was assessed by the results of the hair samples of both volunteers. For the sample of volunteer 1 at least the results of 6 (out of 23) elements were within an acceptable range of +/- 30% from the consensus value (= mean of all seven labs). For volunteer 2 this was only the case for 2 (!) elements. Differences of more than 100% were found for most other elements. Moreover, in the vast majority of the tested elements there was no comparability of the cLassification to the respective reference ranges of the different laboratories. For example, for volunteer 1 only 3 elements (our of 23!) were identically classified by all seven labs. As neither the analytical results nor the classification to the individual reference ranges by the laboratories correspond in tolerable borders, conclusions, drawn from these results, cannot be valid. Hair mineral analysis from these laboratories is unreliable. Therefore we must recommend to refrain from using such analysis to assess individual nutritional status or suspected environmental exposure. PMID- 11878750 TI - Biomonitoring of a worker population exposed to low antimony trioxide levels. AB - Antimony trioxide (Sb2O3) is used as a flame retardant in the textile industry. We carried out a study in a factory for the evaluation of antimony (Sb) occupational exposure and urinary levels in workers exposed to Sb2O3. Urinary levels and airborne Sb2O3 personal exposure values were very low when compared to international occupational standards (500 microg/m3, as Sb). The range of forty two personal exposures was 0.01-0.55 microg Sb/m3, while twenty-four area samplings ranged from < 0.01 microg Sb/m3 to 1.45 microg Sb/m3. The mean urinary Sb levels at the beginning (n = 39) and end of the shift (n = 39) were 0.31 +/- 0.25 microg/L and 0.35 +/- 0.29 microg/L respectively, without any significant statistical difference. When the workers were divided into two subgroups according to "higher" and 'lower" exposure levels, a statistical difference (P < 0.001) was observed between the mean Sb urinary levels of the two subgroups during the workweek, both at the beginning and end of the shift. A statistical difference was also observed between the above mentioned subgroups and the controls (n = 15). No correlation was found between personal Sb2O3 exposure and the difference in urinary Sb levels at the beginning and end of the workshift on the day the flame retardant was utilized. This lack of correlation could be due to low airborne Sb2O3 levels and Sb dietary intake, estimated as 3 microg/day in UK, but not yet fully investigated in Italy. Any accidental occupational Sb per os exposure however low, could further enhance the lack of correlation. PMID- 11878751 TI - Selenite inhibition of Coxsackie virus B5 replication: implications on the etiology of Keshan disease. AB - Keshan disease is a cardiomyopathy of unknown origin reported in some areas of China. Because of epidemiologic features, this disease was ascribed to an infectious agent, likely a Coxsackie virus, but it has also been thought to depend on selenium deficiency, mainly because selenite is effective in its prophylaxis. We examined the hypothesis that pharmacological activity of selenite on Coxsackie virus growth was associated with prevention of Keshan disease. We studied the antiviral effects of three selenium compounds on Coxsackie virus B5 replication: five microM selenite reduced viral replication, whilst 10 microM selenate and selenomethionine did not exhibit any antiviral activity. The inhibitory activity of selenite on viral replication was due to its toxicity following its interaction with thiols, as that activity could be blocked by dithiothreitol, a sulfhydryl-protecting agent known to reverse several toxic effect of selenite. Zinc, another inhibitor of selenite toxicity, also counteracted the antiviral effect of selenite. The selenium compounds showed only limited activity against herpes simplex 1 virus and IHD strain of vaccinia virus. A direct inhibitory effect of selenite on Coxsackie virus replication might explain the efficacy demonstrated by this compound in the prophylaxis of Keshan disease. PMID- 11878752 TI - Parameters of dietary selenium and vitamin E deficiency in growing rabbits. AB - 4 x 5 growing female rabbits (New Zealand White) with an initial live weight of 610 +/- 62 g were fed a torula yeast based semisynthetic diet low in selenium (<0.03 mg/kg diet) and containing <2 mg alpha-tocopherol per kg (group I). Group II received a vitamin E supplementation of 150 mg alpha-tocopherylacetate per kg diet, whereas for group III 0.40 mg Se as Na-selenite and for group IV both supplements were added. Selenium status and parameters of tissue damage were analyzed after 10 weeks on experiment (live weight 2,355 +/- 145 g). Selenium depletion of the Se deficient rabbits (groups I and II) was indicated by a significantly lower plasma Se content (group I: 38.3 +/- 6.23 microg Se/mL plasma, group II: 42.6 +/- 9.77, group III: 149 +/- 33.4, group IV: 126 +/- 6.45) and a significantly lower liver Se content (group I: 89.4 +/- 18.2 microg/kg fresh matter, group II: 111 +/- 26.2) as compared to the Se supplemented groups III (983 +/- 204) and IV (926 +/- 73.9). After 5 weeks on the experimental diets differences in the development of plasma glutathione peroxidase were observed. As compared to the initial status group (45.2 +/- 4.50) pGPx activity in mU/mg protein was decreased in group I (19.1 +/- 7.08), remained almost stable in the vitamin E supplemented group II (46.3 +/- 11.2) whereas an elevated enzyme activity was measured in the Se supplemented groups III (62.4 +/- 23.9) and IV (106 +/- 19.9). In the rabbit organs investigated 10 weeks of Se deficiency caused a significant loss of Se dependent cellular glutathione peroxidase activity (GPx1) of 94% (liver), 80% (kidney), 50% (heart muscle) and 60% (musculus longissimus dorsi) in comparison to Se supplemented control animals. Damage of cellular lipids and proteins in the liver was due to either Se or vitamin E deficiency. However damage was most severe under conditions of a combined Se and vitamin E deficiency. It can be concluded that the activity of plasma glutathione peroxidase is a sensitive indicator of Se deficiency in rabbits. The loss of GPx1 activity indicates the selenium depletion in various rabbit organs. Both selenium and vitamin E are essential and highly efficient antioxidants which protect rabbits against lipid and protein oxidation. PMID- 11878754 TI - Trace mineral status in post menopausal women: impact of hormonal replacement therapy. AB - The risks of disturbances in trace mineral nutrition and metabolism are high following menopause. The aim of the study was to investigate the trace mineral status in postmenopausal women and the influence of hormonal replacement therapy on this status. Forty-four healthy postmenopausal women, aged 50-60 years old participated in the study. Eighteen were treated by combined hormonal replacement therapy (HRT) per os for at least two years, and 26 were untreated. Plasma trace mineral levels (Zn, Se, Cr, Mn, Cu), red blood cell antioxidant enzymes (Cu-Zn SOD, Se-GPX, Cu), urinary Zn, Cr, Mg, and Ca excretion were measured. Zinc, selenium and manganese plasma levels, activities of Cu-Zn-SOD and GSH-Px in erythrocytes were not statistically different between the two groups. The percentage of zinc plasma levels below the cut off of 10.7 micromol/L was higher in HRT treated group than in untreated one, whereas zinc excretion was reduced. Plasma copper concentrations were higher in women treated by HRT, whereas erythrocyte copper levels were not modified. Plasma chromium concentrations were significantly higher in women receiving HRT and urinary Cr excretion was decreased. The HRT group also exhibited lower losses of urinary zinc and magnesium than untreated women. These data suggest that hormonal replacement therapy provides beneficial effects on trace mineral status related to menopause. PMID- 11878753 TI - Distribution of an 18 kDa-selenoprotein in several tissues of the rat. AB - By combining methods for trace element analysis, tracer techniques and various biochemical and electrophoretical procedures, information on the characteristics of an 18 kDa-selenoprotein was obtained. By labeling of rats in vivo with [75Se] selenite and gel electrophoretic separation of the proteins in tissues and subcellular fractions, a larger number of selenium-containing proteins could be distinguished. In most of the tissues investigated a labeled 18 kDa-band was present. After co-electrophoresis of the 18 kDa-bands from kidney, liver and brain we found that they all migrated in the same way. Using ultracentrifugational fractionation the 18 kDa-band was localized in the mitochondrial and microsomal membranes. Two-dimensional electrophoresis showed that it consists of a single selenium-containing protein with an isoelectric point of about 4.9-5.0. By means of proteolytic cleavage of the 18 kDa-protein and separation of its peptides by tricine-SDS-PAGE six selenium-containing peptides with molecular masses of 17, 16, 14, 12, 10, and 8 kDa were detected. After electrophoretic separation of the mitochondrial and/or microsomal proteins and acid hydrolysis of the electroeluted protein its amino acid composition was analyzed by RP-HPLC. In this way it was shown that selenium is present in the 18 kDa-protein in form of selenocysteine which is a characteristic of a genetically encoded selenoprotein. PMID- 11878755 TI - Differences in motility pattern between human buccal fibroblasts and periodontal and skin fibroblasts. AB - Migration of fibroblasts from surrounding normal tissue into the wound bed is an important requirement for successful wound healing. This study investigated the motility pattern of buccal, periodontal and skin fibroblasts to determine whether differences in the wound healing efficiency at these sites can be explained by differences in the motile behavior of their respective fibroblast populations. The migratory characteristics were studied in a two-dimensional culture system. The migration of single cells was time-lapse video recorded at intervals of 15 min for a period of 6 h using a computer-assisted microscope work-station. For evaluation of cell morphology, cell contours were recognized semiautomatically and used for determination of cell area, cell spreading and number and length of processes. We found that the cellular displacement of the buccal fibroblasts was only approximately 50% of the cellular displacement of periodontal and skin fibroblasts. The decreased cellular displacement of the buccal fibroblasts was found to be due to both lower cellular speed and less persistence in direction. The buccal fibroblasts also displayed smaller areas and longer processes. The differences in cellular morphology and motility pattern amongst the three fibroblast types could not be explained by differences in secretion of extracellular matrix components and are therefore believed to reflect phenotypic differences amongst fibroblast subpopulations. PMID- 11878756 TI - Establishment of a convenient system for the long-term culture and study of non neoplastic human salivary gland epithelial cells. AB - Epithelial cells appear to play an important role in the initiation and maintenance of autoimmune lesions in the salivary glands of patients with Sjogren's syndrome. Therefore, the detailed study of immunological function of salivary gland epithelial cells (SGEC) may provide useful information for the understanding of Sjogren's syndrome pathogenesis. In this report we aimed to formulate a protocol for the establishment of human non-neoplastic SGEC lines as a tool for the study of the physiology and pathophysiology of these cells. Pointing towards a practical approach, we sought to establish SGEC lines from quite a limited amount of biopsy tissue obtained during the diagnostic evaluation of patients. Herein, the favorable conditions for the long-term maintenance of human non-neoplastic SGEC lines are presented and involve the successive application of a serum-containing and a serum-free culture medium, supplemented with essential epithelial growth factors. This protocol has been found reliable and convenient, as attested by the reproducible establishment of non-neoplastic SGEC lines. The analysis of SGEC phenotypic features, as well as a coculture system for the study of interactions between epithelial cells and lymphocytes, are also described. Such techniques may provide valuable means for the functional and molecular investigation of human SGEC and particularly for the study of Sjogren's syndrome and other disorders of glandular epithelia. PMID- 11878757 TI - The oral anti-volatile sulphur compound effects of zinc salts and their stability constants. AB - Volatile sulphur compounds (VSC) produced in the oral cavity, are a major cause of oral malodour. Zinc (Zn) ions inhibit VSC formation. The objective of this study was to examine whether Zn salts with low stability constants were more suitable as sources of Zn in lozenges than salts with high stability constants. The former provide free Zn ions upon dissolution in water, whereas the latter provide almost no free Zn. Identical lozenges containing Zn-acetate and gluconate, which have low stability constants, and Zn citrate and amino acid chelated Zn, which have extremely high stability constants, were tested. All the lozenges contained 0.9% w/w Zn. Ten volunteers sucked the lozenges until dissolved, and oral VSC were measured by gas chromatography. Zn-acetate, gluconate and -chelate had an impressive anti-VSC effect even 3 h after the lozenges were taken. Zn citrate had significantly less effect than the other lozenges except Zn acetate after 2 and 3 h. It was concluded that the anti-VSC effect was not related to the stability constants of the Zn compounds tested. Alternative ligands. with stronger affinity for Zn than the ligands in the lozenges, must be present in the oral cavity to explain these results. It is suggested that the sulphide ion may serve this function. PMID- 11878758 TI - Asymmetrical morphogenesis and medio-lateral positioning of molars during mouse development. AB - The functionality of the dentition depends on occlusal relationships between opposing crown surfaces. To investigate the relative changes in positioning of upper and lower molar germs during mouse development, we used serial histological sections of late day 13 (embryonic day (ED)13.5) to early day 18 (ED18) foetus heads and performed computer-aided 3D reconstructions. From ED13.5 to ED15.5. the first lower molar (M1) got a less medial position relative to its upper counterpart (M1); superimposition progressed postero-anteriorly. From ED14.5, the apparent medial displacement of M(1) vs. M1 was partly due to the asymmetrical growth of the M(1) to give rise to the lingual row of cusps, conspicuous at ED17. The superimposition of M(2)/M2 along the medio-lateral axis was observed from their bud stage (ED14.5), and the one of M(1)/M1 was almost complete at ED15.5. However, this was not the final position. as at ED 18, M1 and M2 had a more lateral location than their upper counterparts. Immunostaining showed that differential expression of antigens associated to desmosomes but not to adherens junctions might be involved in the asymmetrical development of M(1) thus contributing to the relative medio-lateral positioning of the first molars at early stages. PMID- 11878759 TI - Effects of resin based dental composites on fertility of male mice. AB - The purpose of this study was to investigate the effect of leached substances from dental composites on the fertility of male mice. Twenty adult male Swiss mice were divided into two groups of 10. Leached substances from composite specimens were administered, intragastrically, daily to the mice in the test group and distilled water to the control group for 28 d. Each mouse from both groups was mated with two untreated females. After mating, fertility of male mice in both groups was assessed. There was a significant reduction in the number of pregnancies in females mated with test males in comparison to those mated with control males (12/20 vs. 18/20, respectively). Females impregnated by test male mice showed a significant increase in the number of resorptions out of the total number of implantations (11% test vs. 1.5% control). The testicular sperm count and daily sperm production of the males in the test group was significantly reduced. Moreover, the relative weights of the testes and seminal vesicles were also significantly reduced in the test group. These results suggest that leached components from resin based dental composite materials have an adverse effect on the fertility and reproductive system of male mice. PMID- 11878760 TI - The effect of dental restorative materials on dental biofilm. AB - To investigate the arrangement of biofilms formed in vivo, volunteers wore splints with slabs of six different dental materials inserted to collect smooth surface plaque. After 5 d of undisturbed plaque accumulation, the specimens were vital stained and analyzed by the confocal laser scanning microscopy (CLSM) to evaluate the percentage of vital biofilm microflora (VF percentage). Further parameters were the area of the specimens covered by plaque (surface coating; SC, %) and the height of the biofilms (BH, pm). The metals amalgam and gold, the compomer, as well as the glass-ionomer cement harboured an almost entirely dead biofilm (VF <8%). Resin composite led to vitality values between 4 and 21%, while a very thin biofilm on ceramic revealed the highest vitality values (34-86%). SC varied from 6% on glass-ionomer cement to 100% on amalgam. BH reached its highest value on amalgam and gold of 17 and 11 microm, respectively, while heights of between 1 and 6 microm were found on the ceramic, resin composite, compomer and the glass-ionomer cement. Within their limits, the present findings indicate that amalgam, gold, compomer and glass-ionomer cement exert an influence against the adhering biofilm. No general relationship could be established between the different parameters VF percentage, SC percentage and BH (microm). PMID- 11878761 TI - Characteristics of tooth wear in relation to different nutritional patterns including contemporary and medieval subjects. AB - The present study sought to evaluate the characteristics of tooth wear in subjects with an acidic diet in relation to subjects with substantially different nutritional patterns. The evaluation included medieval skulls (group 1, n = 102, abrasive nutrition), study models of individuals living on an acidic diet (group 2, n = 100) and randomly selected subjects (group 3, n = 100, average Western diet). Wear was visually recorded on oral, vestibular and occlusal/incisal surfaces using quantitative and morphological criteria. The mean age was 42.3 +/- 15.3 yr in group 1, 40.9 +/- 11.2 yr in group 2 and 36.7 +/- 11.7 yr in group 3. Group 1 exhibited the most pronounced substance loss, followed by groups 2 and 3. On occlusal surfaces, cupping was common in group 1 followed by group 2, but was rare in group 3. An inverse relation was found for facets. On buccal surfaces, no substance loss was observed in group 1, whereas in group 2, 63% had at least one tooth with a buccal lesion compared to 8% in group 3. The occlusal substance loss observed in subjects exposed to acids may be interpreted as increased abrasion/demastication of acid-softened dental hard tissues. The occurrence of concavities on smooth surfaces appears to be significant for the diagnosis of dental erosion. PMID- 11878762 TI - Task-related electromyographic spectral changes in the human masseter and temporalis muscles. AB - The masticatory muscles differ in their fiber type composition. It can therefore be expected that their electromyographic (EMG) power spectra will differ during the performance of different bite force tasks. In the present study, surface EMG activity was picked up from the masseter and from the anterior and posterior temporalis muscles of nine adult subjects. At a bite force level as low as 25 N, the mean power frequency (MPF) values of the posterior temporalis were significantly lower than those of the masseter and anterior temporalis. The MPF values of the masseter muscles decreased with an increase of bite force magnitude, whereas the MPF values of the anterior and posterior temporalis did not change significantly. The MPF values were significantly influenced by the direction of bite force. The observed changes of MPF are possibly related to the recruitment of different fiber types, and support the concept that the masticatory muscles behave heterogeneously. PMID- 11878763 TI - Exploiting molecular mimicry: defining rules of the game. AB - Molecular mimicry has been touted as a mean to develop new generation of vaccines to target carbohydrate antigens on pathogens and on tumor cells. Structural and immunological rules governing molecular mimicry require definition for its successful exploitation. Of interest are the kinds of structures that peptides adopt as carbohydrate mimics, the extent to which topological or sequence similarities among peptide mimeotopes define serum cross-reactivity to carbohydrate antigens and the extent to which peptide mimeotopes affect T-cell responses. Rational design concepts can be applied to define how a peptide may mimic carbohydrate antigens, similarities in binding affinities of antibodies for carbohydrate and for peptides, how peptides can mimic core structures on otherwise dissimilar carbohydrate antigens, and how peptide mimeotopes can be used to manipulate cellular responses not achievable by carbohydrate antigens. PMID- 11878764 TI - Selection of phage-displayed peptides recognised by monoclonal antibodies directed against the lipopolysaccharide of Brucella. AB - Panning and screening of various phage display libraries with monoclonal antibodies (mAbs) directed against the O-chain of the lipopolysaccharide (LPS) of Brucella sp. allowed the identification of peptidic mimotopes of some O-chain epitopes. Four mAbs were tested. The A76-12G12 mAb, which is specific for LPS of all strains of Brucella, either A- or M-dominant, did not yield any peptidic mimotope, despite a specific yield enrichment during the rounds of panning. The B66-4F9 mAb, that recognises an epitope common to both Brucella sp. and Yersinia enterocilitca O:9 strains, allowed the selection of only one phage clone that was shown to be an antigenic but not immunogenic mimotope. The B66-2C8 and A15-6B3 mAbs, respectively, specific for the LPS of A-dominant and M-dominant Brucella sp., yielded several sequences, which allowed the determination of consensus sequences. These consensus will be of high interest for the construction of second generation libraries. For the best binding peptides, competition with LPS for the binding to the mAb is detected, which suggests that the peptides bind to the paratope of the mAb. The phages selected from the libraries were used to immunise mice, and a weak antibody response directed against LPS has been observed for some peptides. These data suggest that a subset of the selected peptides are immunogenic mimotopes of the LPS epitopes. PMID- 11878765 TI - Molecular mimetics of Neisseria meningitidis serogroup B polysaccharide. AB - Strains of Neisseria meningitidis serogroup B (NmB) are an important cause of meningitis and sepsis. Efforts to develop a NmB vaccine have been hampered by poor immunogenicity of the polysaccharide capsule, which cross-reacts with host polysialic acid, and the danger of eliciting autoantibodies. To investigate the potential of molecular mimetics to circumvent these problems, we prepared murine monoclonal antibodies (mAbs) against the N-propionyl derivative (N-Pr) of NmB polysaccharide. Several mAbs were found that reacted with capsular polysaccharide epitopes, which were distinct from host polysialic acid. These mAbs also passively conferred protection against experimental bacteremia. We used these mAbs to screen novel independently folding peptide phage display libraries, and pools of combinatorial small molecules, each consisting of approximately 30 to approximately 700 small molecules of diverse composition. To date, several mimetic candidates have been identified. One is a peptide selected from a library of independently folding alphabeta peptides, and others are peptoid dimers or trimers selected from the small molecule pools. The peptoids contain an indan type of ring system, and some of them also contain a large hydrophobic group such as oleyl amine or dehydroabietyl amine, and a positively charged group at the amino-terminus. Both the alphabeta peptide from the phage library, and the peptoids from the small molecule pools, inhibit binding of the mAbs to N-Pr NmB polysaccharide. Future studies will focus on the structure/activity relationship of these mimetics, and the development of immunogens that may be capable of eliciting anticapsular antibody without autoantibody activity. PMID- 11878767 TI - Strategies for mimicking Neisserial saccharide epitopes as vaccines. AB - Monoclonal antibody (mAb) 2C7 recognizes a conserved and widely expressed oligosaccharide (OS) epitope on Neisseria gonorrhoeae. This OS epitope evokes a significant bactericidal and opsonic immune response after natural infection and vaccination. The OS epitope structure represents an excellent target for a potential protective gonococcal vaccine. Because carbohydrate antigens are T-cell independent, inducing weak antibody responses, OS molecules are not useful immunogens. We developed and examined two different strategies to mimic the 2C7 OS epitope: (i) an anti-idiotope (mAb CA1); and (ii) a peptide (PEP-1). These surrogate immunogens elicited antibody responses in mice (CA1 and PEP-1) and rabbits (CA1) that were bactericidal in vitro against gonococci. Both CA1 and PEP 1 are true immunologic mimics of OS and may form a basis for the development of vaccine candidates for human immunization against N. gonorrhoeae. PMID- 11878768 TI - Development and characterization of anti-idiotype based peptide and DNA vaccines which mimic the capsular polysaccharide of Neisseria meningitidis serogroup C. AB - Anti-idiotypic antibodies and peptides which mimic antigens may offer an alternate strategy for converting a thymus-independent (TI) antigen to a thymus dependent (TD) antigen. We have developed an anti-idiotype based peptide mimic of the capsular polysaccharide of N. meningitidis serogroup C (MCPS) which induces a T-dependent protective immune response in mice. Subsequent studies have demonstrated that immunization of severe combined immunodeficient mice reconstituted with human B cells respond to immunization with the MCPS peptide mimic with bactericidal anti-polysaccharide directed antibody response. We hypothesized that administration of a DNA vaccine resulting in endogenous expression of this carbohydrate peptide mimic would induce anti-MCPS antibodies. PMID- 11878766 TI - Initial studies of the molecular basis of peptide mimicry of group B streptococcal type III capsular polysaccharide. AB - We have previously shown that the peptide FDTGAFDPDWPA is a mimetic of the group B streptococcal type III capsular polysaccharide (CPS(III)). It binds to anti CPS(III) antibodies and can be used to immunize mice and produce anti-CPS(III). In this paper we investigate the molecular mechanisms underlying this peptide carbohydrate mimicry in two ways. First, we have examined the conformation of the peptide by NMR spectroscopy in water. Next, we have produced monoclonal anti peptide and anti-CPS(III) antibodies, compared their fine specificity, and have sequenced them. The results indicate that the peptide assumes a conformation that may be similar to that of the CPS(III) in solution and that the peptide and CPS(III) elicit an overlapping repertoire of antibodies. PMID- 11878769 TI - Anti-idiotypic vaccination against group B streptococci. AB - We describe the antigenic properties of an anti-idiotypic single chain fragment variable (scFv) recombinant antibody mimicking the type III capsular polysaccharide of group B streptococci (GBS), an important cause of neonatal sepsis. This scFv could compete with the nominal antigen for binding to specific mouse or rabbit antibodies. Moreover, the scFv elicited, in mice, the production of antibodies which reacted against the type IlI polysaccharide and passively protected neonatal pups from GBS disease. Maternal immunization with the scFv also protected neonatal mice against GBS infection. Next, the scFv was expressed on the surface of the commensal bacterium Streptococcus gordonii. Intravaginal inoculation of mice with these recombinant bacteria induced significant elevations in serum titers of anti-GBS type III antibodies. Therefore, the expression scFv in commensal bacteria may be a convenient and effective way of delivering anti-idiotypic vaccines. PMID- 11878771 TI - Induction of cross-reactive humoral immune response by immunization with mimotopes of the hypervariable region 1 of the hepatitis C virus. AB - Hepatitis C Virus (HCV) is a major cause of chronic hepatitis, liver cirrhosis and hepatocellular carcinoma, worldwide, and the development of an effective vaccine represents a high priority goal. The Hyper Variable Region 1 (HVR1) of the second Envelope protein (E2) of HCV contains a principal neutralizing determinant, but it is highly variable among different isolates and it is involved in the escape from host immune response. Thus, to be effective, a vaccine should elicit a cross-reacting humoral response against the majority of viral variants. We show that it is possible to achieve a broadly cross-reactive immune response in rabbits by immunization with mimotopes of the HVR1. selected from a specialized phage library using HCV patients' sera. At least some of the cross-reacting anti-mimotope antibodies, elicited in rabbits, recognize discontinuous epitopes in a manner similar to those induced by the virus in infected patients. PMID- 11878770 TI - Recombinant Streptococcus gordonii for mucosal delivery of a scFv microbicidal antibody. AB - The gram-positive bacterium Streptococcus gordonii was engineered to express the microbicidal molecule H6, which is an antiidiotypic single chain antibody mimicking a yeast killer toxin. S. gordonii is a human commensal which we developed as a model system for mucosal delivery of heterologous proteins. The in vivo candidacidal activity of both H6-secreting and H6-surface-displaying streptococcal strains were assayed in a well-established rat model of vaginal candidiasis. At day 21 full clearance of Candida albicans infection was observed in 75% of animals treated with the H6-secreting strain, and in 37.5% of animals treated with the strain expressing H6 on the surface, while all animals treated with the control strain were still infected. The observed candidacidal effect was comparable with that observed with the antimycotic drug fluconazole. These data confirm the potential of H6 as a candidacidal agent and show how promising is the approach of using recombinant bacteria for mucosal delivery of biologically active molecules. PMID- 11878772 TI - T-cell tolerance and autoimmune diabetes. AB - Herein we describe the major signaling events that occur in T-cells upon T-cell receptor (TCR) engagement, and the mechanisms responsible for the induction of T cell anergy that may ultimately lead to the development of immunospecific therapies in T-cell mediated autoimmune diseases. A new type of antigen presenting molecule (dimeric MHC class-II/peptide, DEF) endowed with antigen specific immunomodulatory effects such as induction of Th2 polarization and T cell anergy is also described as a potential antidiabetogenic agent. According to our preliminary results, the MHC II/peptide-based approach may provide rational grounds for further development of antigen-specific immunotherapeutic agents such as human-like MHC lI/peptide chimeras endowed with efficient down-regulatory effects in CD4 T-cell-mediated autoimmune diseases such as Type 1 diabetes, multiple sclerosis, primary biliary cirrhosis, and rheumatoid arthritis. PMID- 11878773 TI - Exploring end-of-life care for children with cancer. PMID- 11878774 TI - Variables influencing end-of-life care in children and adolescents with cancer. AB - BACKGROUND: The purpose of this study was to describe the variables influencing end-of-life care in children and adolescents dying of cancer. MATERIALS AND METHODS: Records of 146 children with cancer who died at Children's Hospital were reviewed for demographics, diagnosis, location of death, withdrawal of life support, use of "do not resuscitate" (DNR) orders, and the length of time that those orders were in effect. RESULTS: Ninety-five patients were evaluated. Fifty nine died of progressive disease and 36 deaths were therapy-related. Sixty-four percent of disease-related deaths occurred at home with support from home care or hospice. Only 10% of all patients died while receiving maximal aggressive support in the intensive care unit. Age, diagnosis (solid tumor vs. leukemia), cause of death, length of last hospital admission, and the duration of DNR orders had a significant correlation with the place of death and referral to and use of hospice. Thirty-five percent of all patients had hospice support. CONCLUSIONS: Most children who die of cancer die because of progressive disease at home with hospice support. Do not resuscitate orders were written for most patients who died. End-of-life decisions are influenced by patient diagnosis, cause of death, and age. PMID- 11878775 TI - Leukemic transformation in patients with severe congenital neutropenia. PMID- 11878776 TI - Glypican-3 expression in Wilms tumor and hepatoblastoma. AB - BACKGROUND: Glypican-3 (GPC3) is a heparan sulfate proteoglycan. When it is disrupted, it causes the X-linked gigantism-overgrowth Simpson-Golabi-Behmel syndrome. Its involvement in growth control is consistent with recent reports that it can bind to growth factors, possibly including insulin-like growth factor 2. Further, it has been hypothesized that it may function as a tumor suppressor gene in breast and ovarian carcinomas and mesotheliomas. PATIENTS AND METHODS: RNA and protein were extracted from Wilms tumor and hepatoblastoma tissue samples and GPC3 levels were measured in these extracts by Northern blotting, reverse transcription polymerase chain reaction, and immunoblotting. RESULTS: In contrast to published results with carcinomas, high levels of GPC3 expression were found in Wilms tumor and hepatoblastoma. Low or undetectable expressions of this gene were found in normal tissue surrounding the tumor. CONCLUSIONS: Increased expression of GPC3 in Wilms tumor and hepatoblastoma suggests a growth-promoting or neutral activity for this gene product rather than a growth-suppressive effect. PMID- 11878777 TI - Buthionine sulfoximine and myeloablative concentrations of melphalan overcome resistance in a melphalan-resistant neuroblastoma cell line. AB - BACKGROUND: Alkylator resistance contributes to treatment failure in high-risk neuroblastoma. Buthionine sulfoximine (BSO) can deplete glutathione and synergistically enhance in vitro sensitivity to the alkylating agent melphalan (L PAM) for many neuroblastoma cell lines, but optimal use of this combination needs to be defined because clinical responses have been less frequent and not durable. PATIENTS AND METHODS: The authors established and characterized a neuroblastoma cell line (CHLA-171) from a patient who died of progressive disease after treatment with BSO and low-dose L-PAM. RESULTS: CHLA-171 lacks MYCN amplification, expresses PGP (P-glycoprotein) 9.5 RNA, and shows cell surface antigen expression (human leukocyte antigen class I weakly positive, but HSAN 1.2 (hybridoma, SAN 1.2) and anti-GD2 (anti-ganglioside GD2 antibody) strongly positive) characteristic of neuroblastoma cell lines. Twenty-four hours of BSO treatment (0-1,000 micromol/L) maximally depleted CHLA-171 glutathione to 36% of baseline. The cytotoxic response of CHLA-171 to BSO and L-PAM, alone and in combination, was measured by digital image microscopy (DIMSCAN) over a range of drug concentrations and compared with drug levels obtained in the patient during BSO/L-PAM therapy. As single agents, CHLA-171 was highly resistant to L-PAM (LD90 = 42 micromol/L; peak plasma concentration in the patient equals 3.9 micromol/L) and moderately resistant to BSO (LD90 = 509 micromol/L; steady-state concentration in the patient equals 397 micromol/L). Treatment with a 10:1 (BSO:L PAM) fixed ratio combination synergistically overcame resistance (3-4 logs of cell kill, combination index <1) at clinically achievable levels of BSO (100-400 micromol/L) and levels of L-PAM (10-40 micromol/L) clinically achievable only with hematopoietic stem cell support. CONCLUSIONS: The in vitro results obtained for CHLA-171 suggest that BSO/L-PAM therapy may be optimally effective for drug resistant neuroblastoma using myeloablative doses of L-PAM. PMID- 11878779 TI - Initial bone marrow aspiration in childhood idiopathic thrombocytopenia: decision analysis. AB - PURPOSE: Bone marrow aspiration (BMA) is routinely performed before starting steroid therapy in children with idiopathic thrombocytopenia, primarily to rule out leukemia. METHODS: A decision tree for the initial management of a child older than age 6 months, presenting with idiopathic thrombocytopenia, without blasts on the peripheral smear was constructed. The three strategies are: 1) initial BMA in all patients; 2) initial BMA only in patients at high risk; and 3) empiric therapy for all patients without initial BMA. High-risk criteria include any of: platelet count >50 x 10(9)/L; hemoglobin <100 g/L (age younger than 12 months) or <110 g/L (age older than 12 months): white blood cell count <5 x 10(9)/L (younger than 6 years) or <4 x 10(9)/L (older than 6 years); or absolute neutrophil count <1.5 x 10(9)/L (younger than 6 years) or <2 x 10(9)/L (older than 6 years). The results are expressed as quality-adjusted life years (QALYs), a measure that estimates the overall life expectancy in years for patients receiving a particular treatment strategy, corrected for the patient's quality of life. RESULTS: The base case results are: 1) BMA all = 69.649 QALYs; 2) high-risk BMA = 69.652 QALYs; and 3) empiric therapy = 69.644 QALYs. These results indicate a three-way toss-up because there is less than a 4-day quality-adjusted difference (0.01) between strategies. CONCLUSION: This study indicates that the initial BMA does not significantly change the overall QALYs of a child presenting with thrombocytopenia and, consequently, is not mandatory in every patient before starting steroids. PMID- 11878778 TI - Effects of granulocyte colony-stimulating factor on neutrophil adhesive molecules in neonates. AB - BACKGROUND: Term and preterm neonates experience quantitative and qualitative neutrophil deficiencies resulting in part from decreased production of granulocyte colony-stimulating factor (G-CSF). In adults, G-CSF improves neutrophil function by up-regulating adhesion molecules. PATIENTS AND METHODS: To evaluate the effects of G-CSF on neonatal neutrophil adhesive phenotypes, cord blood samples were incubated with G-CSF or phosphate-buffered saline and stimulated with N-formyl-methionyl-leucyl-phenylalanine (FMLP), and adhesion molecules were evaluated by flow cytometry. RESULTS: In term and preterm neutrophils, G-CSF incubation increased beta2-integrin expression significantly compared with baseline and to a greater extent than observed in adult neutrophils. With FMLP stimulation, beta2-integrin expression increased even more in the G-CSF group. L-selectin expression decreased after G-CSF incubation and decreased even more with FMLP stimulation in the G-CSF group compared with the phosphate-buffered saline group in term and preterm samples, but not in adult samples. CONCLUSIONS: The data show that G-CSF increases expression of beta2 integrin and decreases expression of L-selectin on unstimulated and stimulated term and preterm neonatal neutrophils in vitro. Further study is required to determine whether G-CSF improves neonatal neutrophil function. PMID- 11878780 TI - Typical and atypical Carney's triad presenting with malignant hypertension and papilledema. AB - This report encourages clinicians to consider a diagnosis of Carney's triad in patients with multifocal gastric stromal sarcoma, extraadrenal paraganglioma (predominantly mediastinal), or pulmonary chondroma. The authors conducted a retrospective 20-year survey at the Hospital for Sick Children and identified two children with Carney's triad. One child, presenting atypically with papilledema and fundal hemorrhages from malignant hypertension and benign intracranial hypertension from chronic iron-deficiency anemia, is the second patient ever to date be described with the complete Carney's triad of neoplasms at diagnosis. Another child presented more typically with gastric stromal sarcoma and pulmonary chondroma without paraganglioma. Carney's triad is a rare differential diagnosis for "idiopathic" hypertension or iron-deficiency anemia from chronic gastrointestinal bleeding. If missed, patients with Carney's triad may have the debilitating physical and mental consequences of chronic iron deficiency and may die of untreated prolonged hypertension and metastatic leiomyosarcoma. PMID- 11878781 TI - Upper airway obstruction-related sleep apnea in a child with thalassemia intermedia. AB - Obstructive sleep apnea can be caused by hypertrophy of tonsils and adenoids or neuromuscular diseases. The authors describe a child with thalassemia intermedia in whom severe obstructive sleep apnea syndrome developed. Computed tomography scanning revealed an obstruction of the nasopharynx resulting from extramedullary hematopoiesis. The child was treated with hydroxyurea and blood transfusions. Relief of symptoms was noted 1.5 months after initial treatment. Extramedullary hematopoiesis causes sleep apnea syndrome in thalassemic patients, and the treatment of hydroxyurea and blood transfusion for extramedullary hematopoiesis should be further studied. PMID- 11878783 TI - Ewing sarcoma of the small intestine. AB - This report describes a rare case of Ewing sarcoma (ES) of the small intestine. The patient was a 9-year-old girl with progressive abdominal distension. Computed tomography showed a large mass in the small bowel. Histopathologic examination of the resected tumor showed ES with typical histologic, immunohistochemical, and ultrastructural features. The tumor recurred in the pelvic cavity 18 months after the original surgery. Molecular study of the recurrent tumor confirmed a diagnostic EWS-FLI1 gene fusion. This patient illustrates the unique occurrence of ES in the small intestine. PMID- 11878782 TI - Hepatitis C infection and hepatocellular carcinoma after treatment of childhood cancer. AB - The results of preliminary reports of childhood cancer survivors with hepatitis C infection (HCV) show that in none of these patients did the disease progress to liver failure or hepatocellular carcinoma (HCC). The authors describe two patients who were diagnosed with HCC more than 20 years after the treatment of childhood acute lymphocytic leukemia. Serologic testing, done at the time HCC was diagnosed, found HCV-directed antibodies, suggesting that chronic HCV infection contributed to the development of the subsequent neoplasm. Identification of infected patients will permit intervention to reduce the risk of progressive liver disease and will also assist in defining the risk of and variables contributing to progressive liver disease. PMID- 11878784 TI - Meeting report of the 2000 International Congress of Pediatric Hematology/Oncology: Shanghai Children's Medical Center, Shanghai, People's Republic of China, October 18-20, 2000. PMID- 11878785 TI - Folate status assessment and folic acid supplements in sickle cell disease. PMID- 11878786 TI - On Dr. Louis K. Diamond's 1932 article and subsequent contributions to erythroblastosis fetalis. PMID- 11878787 TI - Overview of the state of the art of Rh disease: history, current clinical management, and recent progress. PMID- 11878788 TI - Gastrointestinal fistulae. PMID- 11878789 TI - Role of somatostatin-14 and its analogues in the management of gastrointestinal fistulae: clinical data. AB - Somatostatin-14 and its analogue octreotide both exert inhibitory effects on gastrointestinal secretions and may therefore be beneficial in the treatment of gastrointestinal fistulae. There are no studies that have compared these two drugs directly and hence this paper aims to review studies that are available for each drug. There are only six controlled studies that have examined the effects of somatostatin-14 and octreotide on fistula output reduction, three for each drug. All studies compared conservative therapy and the drug in combination with conservative therapy. Of the somatostatin-14 studies, two showed a significant effect on output (p<0.05) and the other demonstrated an output reduction on day 1 that was twice that in the control group (NS). Of the octreotide studies, one showed a significant effect (p<0.01) and the other two showed no effect of the drug on output. No study with either drug has demonstrated an increase in the number of patients that have achieved closure. However, a positive effect on the time to achieve closure has been found. Of the five controlled studies with somatostatin-14, all showed a significant reduction in time to closure. Of the two controlled studies with octreotide, one showed a significant reduction (p=0.002) and the other showed no difference. Due to the limited number of trials, a definitive evaluation of the efficacies of somatostatin-14 and octreotide in the treatment of gastrointestinal fistulae is not possible. However, currently available information seems to suggest a considerable benefit of somatostatin-14 when administered in association with standard conservative treatment, but this needs to be confirmed in a large prospective controlled study. PMID- 11878790 TI - The relevance of gastrointestinal fistulae in clinical practice: a review. AB - Gastrointestinal fistulae most frequently occur as complications after abdominal surgery (75-85%) although they can also occur spontaneously--for example, in patients with inflammatory bowel disease (IBD) such as diverticulitis or following radiation therapy. Abdominal trauma can also lead to fistula formation although this is rare. Postoperative gastrointestinal fistulae can occur after any abdominal procedure in which the gastrointestinal tract is manipulated. Regardless of the cause, leakage of intestinal juices initiates a cascade of events: localised infection, abscess formation and, as a result of a septic focus, fistulae formation. The nature of the underlying disease may also be important, with some studies showing that fistula formation is more frequent following surgery for cancer than for benign disease. Fistula formation can result in a number of serious or debilitating complications, ranging from disturbance of fluid and electrolyte balance to sepsis and even death. The patient will almost always suffer from severe discomfort and pain. They may also have psychological problems, including anxiety over the course of their disease, and a poor body image due to the malodorous drainage fluid. Postoperative fistula formation often results in prolonged hospitalisation, patient disability, and enormous cost. Therapy has improved over time with the introduction of parental nutrition, intensive postoperative care, and advanced surgical techniques, which has reduced mortality rates. However, the number of patients suffering from gastrointestinal fistulae has not declined substantially. This can partially be explained by the fact that with improved care, more complex surgery is being performed on patients with more advanced or complicated disease who are generally at higher risk. Therefore, gastrointestinal fistulae remain an important complication following gastrointestinal surgery. PMID- 11878792 TI - Pancreatic surgical complications--the case for prophylaxis. AB - Pancreaticoduodenectomy (Whipple's procedure) represents a considerable surgical challenge. Postoperative complications are common and typically related to leakage of pancreatic exocrine secretions following anastomosis failure. Pancreatic proteases and lipase leaking from the organ remnant attack the surrounding tissue, potentially leading to severe inflammation, tissue necrosis, and fistula formation. In addition, the soft consistency of the normal pancreas can lead to difficulties in manipulating the organ and reduce the integrity of sutures. Pancreatic fistula is the most serious postoperative complication and especially common following resectional surgery for malignant disease. Through prophylactic inhibition of digestive secretions, it should be possible to reduce postoperative morbidity after pancreatic surgery. One such inhibitor is somatostatin-14, an endogenous peptide hormone with pronounced effects on secretion of pancreatic enzymes and hormones, gastrointestinal secretions, and pancreatic blood flow, all of which may decrease the risk of postoperative complications. A limited number of randomised controlled trials have investigated prophylactic administration of somatostatin-14 and the synthetic somatostatin analogue octreotide in reducing complications following pancreatic surgery. While the majority of studies with octreotide demonstrated a significant reduction in the overall complication rate, the benefits appeared less marked in relation to events specifically related to pancreatic secretion. However, preliminary results from a limited number of trials with somatostatin-14, administered as a continuous intravenous infusion, suggest that prophylactic pharmacotherapy produces a significant decrease in fistula formation and secretion related events after pancreaticoduodenectomy. Due to these promising data, further investigation of the role of somatostatin-14 prophylaxis in pancreatic surgery is warranted in large well controlled trials. PMID- 11878791 TI - Optimising the treatment of upper gastrointestinal fistulae. AB - A three stage strategy is generally employed in the management of gastrointestinal fistulae which can form due to surgery, disease, or trauma. The condition is investigated leading to diagnosis, conservative treatment is initiated to stabilise the patient, followed by specific surgical treatment measures in complicated cases, or in the absence of spontaneous closure. Conservative management of fistulae is based on parenteral nutrition and bowel rest, as well as on control of infection, electrolytic disturbances, and local care of the fistula tract. Surgical treatment may be required although generally only in particularly serious cases. Somatostatin-14 has been used in addition to parenteral nutrition to further reduce the volume and enzymatic activity of the fluid output through the fistula tract, generally with good results. The majority of reports have shown a beneficial effect, and randomised studies have demonstrated a reduction in closure time and morbidity. However, due to a combination of the seriousness and rarity of the condition and the difficulties inherent in trial design, data from large scale, double blind, randomised, controlled studies investigating the use of pharmacotherapy in the treatment of established gastrointestinal fistulae are lacking. Nevertheless, preliminary data from initial trials suggest that somatostatin-14 and its analogue octreotide considerably improve the conservative treatment of gastrointestinal fistulae in the absence of distal obstruction. In addition, reduction of the concentration of caustic enzymes in the discharge will benefit both wound healing and nutritional losses. With reduced closure time, the period of hospitalisation will be shortened with potentially considerable economic reductions and improvements in quality of life for the patient. PMID- 11878793 TI - A method of reversible biomolecular immobilization for the surface plasmon resonance quantitative analysis of interacting biological macromolecules. AB - This article presents a new procedure for the immobilization of macromolecules on gold surfaces, with the purpose of studying macromolecular interactions by simple optical configurations rendering surface plasmon resonance. Gold surfaces were covered by a three-layer structure composed of poly-L-lysine irreversibly bound to gold, followed by a second layer of heparin and a third layer of polylysine. The three-layer structure of polylysine-heparin-polylysine remains irreversibly bound to gold, it prevents biomolecules from coming into direct contact with the metal surface, and it allows the irreversible binding of different proteins and polynucleotides. After binding of a macromolecule to the three-layer structure, the interaction with a second macromolecule can be studied, and then the complex formed by the two interacting macromolecules, together with the second heparin layer and the third polylysine layer, can be broken down just by treatment with an alkaline solution having a pH value above the pK value of the amino groups of polylysine. The first polylysine layer remains irreversibly bound to gold, ready to form a new three-layer structure and, therefore, to support a new macromolecular interaction on the same regenerated surface. Polynucleotide interactions, the proteolytic action of chymotrypsin, and the interaction between the component subunits of a heterotetrameric enzyme are described as examples of macromolecular interactions studied by using this system. The method may be especially suitable for developing of low-cost systems aimed to look for surface resonance signals, and it offers the advantage of allowing calculation of parameters related to the size and stoichiometry of the interacting macromolecules, in addition to the kinetic and equilibrium properties of the interaction. PMID- 11878795 TI - A microplate reader-based nonisotopic histone deacetylase activity assay. AB - Recent years have brought an enormous increase in knowledge concerning the involvement of histone deacetylase (HDAC) in gene regulation and the potential use of its inhibitors in transcription therapy. This also stimulates research toward new methods for the determination of HDAC activity and thus the potency of potential inhibitors. We have previously succeeded in developing a nonisotopic assay for HDAC using a fluorescent coumarin derivative of epsilon-acetyllysine. Here we present plate reader-based quantitation as an alternative means for the determination of substrate conversion. A new validated assay procedure with a boradiazaindacene (BODIPY 530/550) rather than a coumarin internal standard was established to allow for fluorescence measurement without chromatographic separation. The method is equal in its sensitivity, accuracy, and precision to the previously published HPLC method. A comparison with a new commercially available homogeneous plate reader assay leads to similar inhibition constants for the HDAC inhibitor trichostatin A. The commercial assay has a higher throughput but its procedure for the detection of HDAC activity could not be applied to our enzyme preparation, while our substrate is also converted by HeLa HDAC. This indicates a broader range of potential applications for our system. PMID- 11878794 TI - Pretreatment procedure for the microdetermination of chondroitin sulfate in plasma and urine. AB - A new, simple, and rapid pretreatment method for the determination of chondroitin sulfate, dermatan sulfate, and hyaluronan from urine and blood plasma samples has been developed. Plasma proteins were first converted into small peptides by digestion using a nonspecific protease, actinase E, and the resulting small peptides were removed by centrifugal filtration. The retained, residual crude glycosaminoglycans, including chondroitin/dermatan sulfates and hyaluronan, were converted into unsaturated disaccharides through the action of chondroitin sulfate lyses. Next, these disaccharides were recovered and purified using centrifugal filtration together with DeltaDi-UA2S, added as an internal standard. The filtered disaccharide mixture was analyzed by HPLC with fluorometric postcolumn derivatization using 2-cyanoacetamide as a fluorogenic reagent. This method was applied to a pharmacokinetic study of chondroitin sulfate administered intravenously to mice. The half-life of the administered chondroitin sulfates, having molecular masses from 6 to 50 kDa, varied depending on their molecular sizes. This new method should be useful for studies on the metabolic fate of exogenously administered glycosaminoglycans in small experimental animals. PMID- 11878796 TI - Determination of RNase A/2'-cytidine monophosphate binding affinity and enthalpy by a global fit of thermal unfolding curves. AB - A spectropolarimeter was used to measure the thermal response curves of RNase A in the presence and absence of the ligand cytidine 2'-monophosphate. A coupled equilibrium model was used to describe the dissociation of the protein-ligand complex (NL<==>N + L) and the thermal unfolding of the free protein (N<==>U). The unfolding curves of the protein in the presence of several different concentrations of ligand were fit to this coupled equilibrium model using global linkage analysis. The best-fitted values for the thermal unfolding of the apo protein were 60.9 +/- 0.2 degrees C (T(m)) and 105.5 +/- 1.4 kcal/mol (DeltaH), while the fitted values for the dissociation of the protein-ligand complex were 1.6 +/- 0.4 microM (K(D)) and 18.7 +/- 1.0 kcal mol(-1) (DeltaH(L)). These values were in excellent agreement with values obtained by other methods. The sensitivity of the data fitting was compared using linear or quadratic temperature dependence for the response curves of the free ligand (L), native apo protein (N), native ligand-bound protein (NL), and unfolded apo-protein (U). There was no significant improvement in the precision of the fitted data when the temperature-dependent response for each species (N, L, NL, and U) was expressed as quadratic functions of temperature. PMID- 11878797 TI - Phospholipid composition of cell-derived microparticles determined by one dimensional high-performance thin-layer chromatography. AB - Microparticles in the circulation activate the coagulation system and may activate the complement system via C-reactive protein upon conversion of membrane phospholipids by phospholipases. We developed a sensitive and reproducible method to determine the phospholipid composition of microparticles. Samples were applied to horizontal, one-dimensional high-performance thin-layer chromatography (HPTLC). Phospholipids were separated on HPTLC by chloroform:ethyl acetate:acetone:isopropanol:ethanol:methanol:water:acetic acid (30:6:6:6:16:28:6:2); visualized by charring with 7.5% Cu-acetate (w/v), 2.5% CuSO(4) (w/v), and 8% H(3)PO(4) (v/v) in water; and quantified by photodensitometric scanning. Erythrocyte membranes were used to validate the HPTLC system. Microparticles were isolated from plasma of healthy individuals (n = 10). On HPTLC, mixtures of (purified) phospholipids, i.e., lysophosphatidylcholine, phosphatidylcholine (PC), sphingomyelin (SM), lysophosphatidylserine, phosphatidylserine, lysophosphatidylethanolamine, phosphatidylethanolamine (PE), and phosphatidylinositol, could be separated and quantified. All phospholipids were detectable in erythrocyte ghosts, and their quantities fell within ranges reported earlier. Quantitation of phospholipids, including extraction, was highly reproducible (CV < 10%). Microparticles contained PC (59%), SM (20.6%), and PE (9.4%), with relatively minor (<5%) quantities of other phospholipids. HPTLC can be used to study the phospholipid composition of cell-derived microparticles and may also be a useful technique for the analysis of other samples that are available only in minor quantities. PMID- 11878798 TI - Characterization and direct quantitation of ceramide molecular species from lipid extracts of biological samples by electrospray ionization tandem mass spectrometry. AB - A rapid, simple, and reliable method has been developed for the characterization and quantitation of ceramide molecular species directly from chloroform extracts of biological samples by electrospray ionization tandem mass spectrometry (ESI/MS/MS). By exploiting the differential fragmentation patterns of deprotonated ceramide ions, individual 2-hydroxy and nonhydroxy ceramide molecular species were readily identified by ESI/MS/MS with the neutral loss of fragments of mass 256.2 and 327.3 which correspond to sphingosine derivatives. The ions generated from the neutral loss of 256.2 (i.e., [M - H - 256.2](-)) are unique for ceramides with N-acyl sphingosine with the 18-carbon homolog. However, the sensitivity for nonhydroxy ceramides in ESI/MS/MS with the neutral loss of 256.2 is approximately threefold higher than that for 2-hydroxy ceramides. The ions resulting from the neutral loss of 327.3 (i.e., [M - H - 327.3](-)) are specific for 2-hydroxy ceramides. Additionally, all ceramides including both 2 hydroxy and nonhydroxy forms can be confirmed and accurately quantitated by ESI/MS/MS with the neutral loss of 240.2 after correction for (13)C isotope factors. This methodology demonstrated a 1000-fold linear dynamic range and a detection limit at the subfemtomole range and was applied to directly quantitate ceramide molecular species in chloroform extracts of biological samples including brain tissues and cell cultures. PMID- 11878799 TI - Cell hybridization by electrofusion on filters. AB - Electric field pulses induce permeabilization and associated fusogenicity in cell membranes. Electrofusion of cells is usually performed in two steps: the first is the creation of close intercellular contacts; the second is an application of electric pulses that induces membrane fusion. Very large cell contacts can be obtained by a filter aspiration method. A cell monolayer is created by controlled suction on biocompatible filter. No spontaneous fusion results. Just after filtration, electrofusion is obtained by field pulses applied parallel to the filter. Cell viability is not strongly affected and cells recover their spherical shape in the minute time range after filtration. The electrical parameters, the cell density, and the flow rate control fusion. Fusion is obtained with cells of different origins with very different adhesion properties. Hybrid cells are easily formed. This approach appears to be a very efficient method for cell hybridization with an easy-to-use protocol. PMID- 11878800 TI - Comparison of the potentiometric, (31)P NMR, and zero-point titration methods of determining ionized magnesium in erythrocytes. AB - A simple and rapid method of determining ionized magnesium in erythrocytes using a potentiometric clinical analyzer, Microlyte 6 (Kone, Finland), was investigated. The erythrocyte cell membranes were destroyed using ultrasound. The results were compared with those obtained with the (31)P nuclear magnetic resonance spectroscopy method and the zero-point titration method using atomic absorption spectrometry. The results obtained from potentiometry and from the other methods did not differ significantly (Student t test, alpha = 0.01). Total magnesium concentration was determined using atomic absorption spectrometry. PMID- 11878801 TI - Immunoprecipitation of DNA-protein complexes cross-linked by cis diamminedichloroplatinum. AB - For the study of in vitro and in vivo DNA-protein interactions, cross-linking reactions driven by UV or formaldehyde have been frequently used, followed by standard protocols of immunoprecipitation and analysis of the DNA isolated from the complexes. Here we present a basically modified method to analyze the DNA protein cross-linked complexes obtained by an alternative cross-linking reagent. The innovations presented here include cross-linking by cis diamminedichloroplatinum II, a fast method to isolate DNA-protein complexes using gel-filtration chromatography, and a modified procedure to obtain specific immunocomplexes that can be analyzed either for DNA or for protein content. The application of this method to two nuclear proteins from chicken liver nuclei is described. PMID- 11878802 TI - Immunodetection of small o-phenylenebismaleimide-labeled peptides through carrier protein display on polyvinylidene fluoride. AB - Protein modification and peptide analysis are important techniques for the elucidation of the structure and function of enzymes. We describe a new technique for the identification of peptides covalently modified with the maleimide cross linker o-phenylenebismaleimide (OPBM). The method can identify labeled peptides without the use of sophisticated instrumentation or radioactive markers and takes advantage of the separating power of RPLC and of the sensitivity of immunoblotting. Chloroplast ATPase F1 was labeled at a single cysteine residue by OPBM and trypsinized. Fractions collected by RPLC were bound to polyvinylidene fluoride (PVDF). Despite the small size of the OPBM-labeled peptide (1.84 kDa) it was possible to immobilize it on PVDF by using glutaraldehyde to conjugate the peptide to a larger, unlabeled protein. Polyclonal antibodies raised against the cross-linker N,N',1,5-naphthalenebismaleimide (NBM) cross-react with OPBM. These antibodies detected the presence of OPBM displayed on the PVDF and correctly identified the RPLC fraction containing the OPBM-labeled peptide as verified by both mass spectroscopy and radiolabeling of OPBM. This method could be adapted to detect the presence of linear epitopes recognized by an antibody and is a broadly applicable technique for the immunodetection of peptides. PMID- 11878803 TI - Attention to relative response across sequential electrodes improves quantitation of coulometric array. AB - HPLC separations coupled with coulometric electrode-based detectors offer potential to evaluate complex biological samples, but analytical approaches to such samples are still being defined. Coulometric electrode array systems use response ratio (as ratio conformity), for quantitation and qualitative characterization. We investigated the influence of changing the ratio conformity requirement (ratio window, RW) on quantitative consistency of the system. Sixty one serum metabolites were analyzed in three cohorts of ad libitum/dietary restricted rats with two RW settings. RW 30 uses channels with <30% ratio variation (vs standard) to quantitate, whereas RW 100 generally uses all channels. Quantitation between the two approaches differed in 29/671, 61/976, and 264/976 metabolites in cohorts A, B, and C, respectively. The changes were 23.0 +/- 16.6, 31.9 +/- 28.6, and 23.6 +/- 17.1% (+/- SD). Correlation analysis showed that 11, 7, and 21% of the 61 metabolites had r(2) < 0.9 in cohorts A, B, and C, respectively. Inspection of HPLC chromatograms revealed that most of the metabolites with changed levels possessed common characteristics (e.g., shoulder peaks, low signal/noise ratios, coeluting peaks). Analysis demonstrated that RW 30 provided precise quantitation, whereas RW 100 did not. These data suggest that narrow ratio window increases quantitative accuracy in coulometric electrode array analysis of the metabolome(s) of complex biological samples (e.g., sera, mitochondria). PMID- 11878804 TI - Tandem mass spectrometric assay for the determination of carnitine palmitoyltransferase II activity in muscle tissue. AB - Carnitine palmitoyltransferase II (CPT-II) mediates the import of long-chain fatty acids into the mitochondrial matrix for subsequent beta-oxidation. Defects of CPT-II manifest as a severe neonatal hepatocardiomuscular form or as a mild muscular phenotype in early infancy or adolescence. CPT-II deficiency is diagnosed by the determination of enzyme activity in tissues involving the time dependent conversion of radiolabeled CPT-II substrates (isotope-exchange assays) or the formation of chromogenic reaction products. We have established a mass spectrometric assay (MS/MS) for the determination of CPT-II activity based on the stoichiometric formation of acetylcarnitine in a coupled reaction system. In this single-tube reaction system palmitoylcarnitine is converted by CPT-II to free carnitine, which is subsequently esterified to acetylcarnitine by carnitine acetyltransferase. The formation of acetylcarnitine directly correlates with the CPT-II activity. Comparison of the MS/MS method (y) with our routine spectrophotometric assay (x) revealed a linear regression of y = 0.58x + 0.12 (r = 0.8369). Both assays allow one to unambiguously detect patients with the muscular form of CPT-II deficiency. However, the higher specificity and sensitivity as well as the avoidance of the drawbacks inherent in the use of radiolabeled substrates make this mass spectrometric method most suitable for the determination of CPT-II activity. PMID- 11878805 TI - Development and applications of surface plasmon resonance-based von Willebrand factor-collagen binding assay. AB - Von Willebrand factor (vWf) functions both as a carrier of factor VIII (fVIII) in plasma and as an adhesive protein providing the primary link between collagen of the extracellular matrix and platelets sequestered from blood flow. The functional activity of vWf correlates with the level of its binding to collagen, which is commonly measured in the enzyme-linked immunosorbent assay (ELISA). We developed an automated collagen-binding assay employing the surface plasmon resonance (SPR) phenomenon, which allows one to quantitatively measure the binding of purified vWf and vWf-containing therapeutic fVIII concentrates to collagen type III immobilized on a biosensor chip. The results of the SPR-based assay highly correlated (r = 0.987) with collagen-binding ELISA. The advantages of the SPR-based assay are its higher accuracy and reproducibility in comparison with ELISA. We applied the developed assay for monitoring structural changes in the vWf component of plasma-derived fVIII/vWf concentrates during a virus inactivation procedure performed by heat treatment. We determined the critical residual moisture content of 2% that can be present in lyophilized concentrates during heat-treatment procedures without causing deteriorative changes in vWf properties. Our data suggest that the SPR-based assay is a useful tool in the development of industrial virus-inactivation procedures, allowing one to preserve vWf activity and achieve the maximal therapeutic efficacy of fVIII/vWf concentrates. PMID- 11878806 TI - Characterization of the H- and L-subunit ratios of ferritins by sodium dodecyl sulfate-capillary gel electrophoresis. AB - Sodium dodecyl sulfate-capillary gel electrophoresis (SDS-CGE) was used to characterize the H- and L-subunit ratios of several mammalian ferritins and one bacterioferritin. Traditionally, SDS-PAGE has been used to characterize the H- and L-subunit ratios in ferritin; however, this technique is relatively slow and requires staining, destaining, and scanning before the data can be processed. In addition, the H- and L-subunits of ferritin are fairly close in molecular weight (approximately 21,000 and approximately 20,000, respectively) and are often difficult to resolve in SDS-PAGE slab gels. In contrast, SDS-CGE requires no staining or destaining procedures and the peak quantitation is superior to SDS PAGE. SDS-CGE is effective in quickly resolving the H- and L-subunits of ferritins from horse spleen, human liver, recombinant human H and L homopolymers, and mixtures of the two- and the single-subunit of a bacterioferritin from Escherichia coli. The technique has also proven useful in assaying the quality of the protein sample from both commercial and recombinant sources. Significant amounts of low-molecular-weight degradation products were detected in all commercial sources of horse spleen ferritin. Most commercial horse spleen ferritins lacked intact H-subunits under denaturing conditions. PMID- 11878807 TI - A continuous fluorimetric assay for tumor necrosis factor-alpha converting enzyme. AB - Fluorogenic peptide substrates with fluorophore/quencher-capped ends have found extensive use in monitoring protease activity in the screening of small-molecule libraries for protease inhibitors. We report here the identification and characterization of a fluorogenic substrate for tumor necrosis factor-alpha converting enzyme (TACE). This substrate is a 10-amino-acid peptide (LAQAVRSSSR) capped with an o-aminobenzoyl group on the N-terminal end and with a 3-(2,4 dinitrophenyl)-L-2,3-diaminopropionic amide group on the C-terminal end. Exhaustive enzymatic conversion of the substrate to products resulted in a fluorescence enhancement of -11-fold. A single cleavage occurred at the A-V scissile bond of the peptide. The validity of this fluorimetric assay for TACE was corroborated by an independent HPLC method. Interestingly, the hydrolysis of the substrate displayed positive cooperativity with a Hill coefficient of 1.5, while the hydrolysis of the corresponding uncapped peptide displayed Michaelis Menten kinetics. A k(cat) value of 21.6 s(-1) and an S(0.5) value of 342 microM were obtained for the fluorogenic substrate. The addition of the two capping groups on the two ends of the peptide enhanced the k(cat) value by 64-fold. Nine additional decapeptides that contained the same capping groups on the two ends and substitutions at the P1 and P1' sites were also tested. TACE appears to slightly prefer the A-V scissile bond. The enzyme also cleaves scissile bonds such as F-V, A-I, and A-L efficiently. PMID- 11878808 TI - Immunoglobulin G-class mouse monoclonal antibodies to major brain gangliosides. AB - Mice genetically engineered to lack complex gangliosides are improved hosts for raising antibodies against those gangliosides. We report the generation and characterization of nine immunoglobulin G (IgG)-class monoclonal antibodies (mAbs) raised against the four major brain gangliosides in mammals. These include (designated as ganglioside specificity-IgG subclass) two anti-GM1 mAbs (GM1-1, GM1-2b), three anti-GD1a mAbs (GD1a-1, GD1a-2a, GD1a-2b), one anti-GD1b mAb (GD1b 1), and three anti-GT1b mAbs (GT1b-1, GT1b-2a, GT1b-2b). Each mAb demonstrated high specificity, with little or no cross-reactivity with other major brain gangliosides. Enzyme-linked immunosorbent assay (ELISA) screening against 14 closely related synthetic and purified gangliosides confirmed the high specificity, with no significant cross-reactivity except that of the anti-GD1a mAbs for the closely related minor ganglioside GT1a alpha. All of the mAbs were useful for ELISA, TLC immunooverlay, and immunocytochemistry. Neural cells from wild-type rats and mice were immunostained to differing levels with the anti ganglioside antibodies, whereas neural cells from mice engineered to lack complex gangliosides (lacking the ganglioside-specific biosynthetic enzyme UDP GalNAc:GM3/GD3 N-acetylgalactosaminyltransferase) remained unstained, demonstrating that most of the mAbs react only with gangliosides and not with related structures on glycoproteins. These mAbs may provide useful tools for delineation of the expression and function of the major brain gangliosides and for probing the pathology of anti-ganglioside autoimmune diseases. PMID- 11878809 TI - Importance of hyaluronan length in a hyaladherin-based assay for hyaluronan. AB - Specific hyaladherin-based assays have been set up to measure the concentration of hyaluronan in biological fluids. Hyaluronectin (HN; a hyaladherin extracted from ovine brain) binds to hyaluronan (HA) that must be 10 units (HA10) or more long. It was therefore of interest to determine whether HN would continue to bind to HA10 in full-length HA since conformational changes might mask potential binding sites. We used the enzyme-linked sorbent assay (ELSA) to assay HA and hyaluronan-derived oligosaccharides, with different standard HAs, and the results were compared to results obtained with the carbazole technique. Oligosaccharide length was calculated from the ratio glucuronic acid/reducing N-acetylglucosamine in fractions of hyaluronidase-digested macromolecular hyaluronan prepared by chromatography; the size of the HA12 oligosaccharide was confirmed by matrix assisted laser desorption ionization mass spectrometry. During the digestion of macromolecular HA with hyaluronidase, the binding of HN to HA first increased and then decreased as shown using the ELSA. The concentration of HA fragments of HA60 and below was overestimated when intact macromolecular HA was used as the reference for the ELSA, while the concentration of HA100 and above was underestimated when HA10 was used as the reference. The binding of HN to HA20, HA40, and HA60 saccharides was consistent with binding to multiples of HA10 sites. In conclusion, the level of HN binding is determined by the conformation of HA, which may mask binding sites. Hence, calibration HA used in the ELSA must be adapted to the size of HA to assay. PMID- 11878810 TI - A direct continuous spectrophotometric assay for glycosidases with 3-nitro-2 pyridyl glycosides by tautomerization of 2-hydroxy-3-nitropyridine. AB - Two kinds of 3-nitro-2-pyridyl glycosides were synthesized and evaluated as substrates for continuous spectrophotometric assay for glycosidases. The liberated aglycon, 2-hydroxy-3-nitropyridine, immediately tautomerized to 3-nitro 2(1H)-pyridone, causing an absorption shift of ca. 60 nm even under acidic conditions (pH 3-6). Consequently, the enzymatic hydrolysis of these glycosides was monitored continuously in the acidic to neutral pH range (pH 4-7), the optimum pH for most glycosidases. The absorbance of liberated aglycon increased linearly at 390 nm until 10% consumption of the substrate to enable the initial rate to be determined at once without terminating the reaction. The kinetic parameters for the hydrolysis of 3-nitro-2-pyridyl glycosides were obtained from the slopes of the progress curves and were compared with those obtained from the conventional discontinuous assay using p- and o-nitrophenyl glycosides as substrates. The kinetic parameters indicated that 3-nitro-2-pyridyl glycosides were more activated and specific substrates, but with less affinity to the enzymes than the corresponding nitrophenyl glycosides. Moreover, the absorbance shift by tautomerization should promise further applications to continuous spectrophotometric assays for other enzymes acting under acidic conditions, such as acid proteases and acid phosphatases. PMID- 11878812 TI - Enzymatic synthesis and purification of aromatic coenzyme a esters. AB - Two recombinant His-tagged proteins, a plant 4-coumarate:coenzyme A ligase (EC 6.2.1.12) and a bacterial benzoate:coenzyme A ligase (EC 6.2.1.25), were expressed in Escherichia coli and purified in a single step using Ni-chelating chromatography. Purified enzymes were used to synthesize cinnamoyl-coenzyme A (CoA), p-coumaroyl-CoA, feruloyl-CoA, caffeoyl-CoA, and benzoyl-CoA. Conversions up to 95% were achieved. Using a rapid solid-phase extraction procedure, the target CoA esters were isolated with yields of up to 80%. Structures were confirmed by analytical comparison with chemically synthesized reference compounds and electrospray ionization-mass spectrometry. The recombinant enzymes were stable for several months at -80 degrees C, thus providing a reliable and facile method to produce these delicate biological intermediates. PMID- 11878811 TI - High-efficiency solid-phase capture using glass beads bonded to microcentrifuge tubes: immunoprecipitation of proteins from cell extracts and assessment of ras activation. AB - We have bonded glass microbeads (425-600 microm diameter) to the inner walls of polypropylene microcentrifuge tubes. In addition to increasing the surface area of the tubes manyfold, the beads provide surface Si groups which can be reacted with a silane compound such as aminopropyltriethoxysilane, yielding a free amino group. The amino group is reacted with another cross-linking reagent, for example, the homobifunctional compound dimethyl suberimidate, which can form a covalent bond with amine groups of proteins. After binding protein A or G to the dimethyl suberimidate, the beads were used to immunoprecipitate proteins from cell extracts; we show that the protein A/G-coated glass beads yield similar amounts of immunoprecipitated proteins as a standard method using protein A- or G agarose beads, but with fewer contaminating proteins. In addition, we show that when immunoprecipitating Ras from cell extracts and measuring the amounts of Ras bound GTP and GDP, the new method yielded higher guanine nucleotide levels than protein G-agarose beads, suggesting that it caused less denaturation of Ras. Because the glass beads are bonded to the walls of the tubes, the immunoprecipitates can be washed rapidly and efficiently, and we show that 20-30 tubes can be washed in 1/10 the time required to wash immunoprecipitates on protein A- or G-agarose beads. PMID- 11878813 TI - Lens organelle degradation. PMID- 11878814 TI - Heterotrimeric GTP-binding proteins in the lacrimal acinar cell endomembrane system. AB - Secretagogues accelerate traffic in the lysosomal and basal-lateral pathways, as well as in the regulated apical secretory pathway, of lacrimal acinar cells. It has been proposed that alterations of protein segregation in compartments where these traffic pathways intersect may influence autoimmune responses. Heterotrimeric GTP-binding proteins couple secretagogue receptor ligand binding to activation of intracellular signaling cascades, but they are also suggested to participate in endomembrane traffic phenomena. Distributions of G(o), G(i3), G(q), G(11), and two G(s)isoforms were mapped in reconstituted lacrimal acini by confocal immunofluorescence microscopy and in lysates of the reconstituted acini by analytical subcellular fractionation. All G proteins examined were detected at low levels in isolated compartments (blm(i,j)) believed to represent the basal lateral plasma membrane. G(i3), G(11), and the G(s)isoforms were concentrated in a series of isolated compartments believed to be related to domains of a basal lateral endosome with sorting and recycling functions (ble-s/r(i,j,k)), a distinct endosomal compartment with basal-lateral membrane-like composition (e blml), and domains of the trans-Golgi network believed to be involved in traffic to and from the basal-lateral membrane (tgn-blmr). G(o)and G(q)were concentrated in compartments believed to represent a mixture of immature and mature secretory vesicle membranes (isvm and svm) and domains of the trans-Golgi network compartment believed to mediate traffic to secretory vesicles (tgn-svr) and to pre-lysosomes (tgn-lr). Confocal fluorescence microscopy confirmed the presence of both basal-lateral membrane and intracellular pools of the G proteins. Stimulation with 10 microM carbachol for 20min caused a component of the G(o)to redistribute away from the isvm+svm; components of the G(i3), G(q), and G(s)to redistribute away from the tgn-svr+tgn-lr; and a component of the G(i3)to redistribute away from the ble-blml+tgn-blmr. Thus, these proteins may participate in endomembrane traffic steps activated by cholinergic stimulation in addition to playing their classical roles in plasma membrane signal transduction. PMID- 11878816 TI - Stimulation of L-type Ca(2+) channels by increase of intracellular InsP3 in rat retinal pigment epithelial cells. AB - The purpose of this study was to investigate the role of voltage-dependent L-type Ca(2+)channels in intracellular Ca(2+)signaling of the retinal pigment epithelium (RPE). Patch-clamp techniques in conjunction with measurements of the intracellular free Ca(2+)using the Ca(2+)-sensitive fluorescence dye fura-2 were performed using cultured rat RPE cells. Intracellular application of inositol 1,4,5-trisphosphate (InsP3; 10 microM) via the patch-pipette during the whole cell configuration led to an increase in the intracellular free Ca(2+)([Ca(2+)](i)). This effect could be reduced by the L-type Ca(2+)channel blocker nifedipine (2 microM). At the moment of the maximal rise in [Ca(2+)](i)L type currents displayed an increase in the current density and shifts in the activation curve and of the steady-state inactivation. Comparable changes of L type channel activity could be observed by induction of capacitative Ca(2+)entry, a maneuver to release Ca(2+)from intracellular Ca(2+)stores independently from InsP3. The increase in L-type Ca(2+)channel activity and [Ca(2+)](i)by intracellular application of InsP3 or induction of capacitative Ca(2+)entry could be inhibited by blocking tyrosine kinase activity using genistein (5 microM) or tyrphostin 51 (10 microM). It is concluded that L-type Ca(2+)channels are involved in the Ca(2+)/InsP3 second messenger system by generating an influx of extracellular Ca(2+)into the cell. This is enabled by depletion of cytosolic Ca(2+)stores and tyrosine kinase-dependent activation of L-type channels. PMID- 11878815 TI - Oral flavonoids, chromocarb diethylamine salt and cyaninosides chloride, to eliminate lipoperoxidation postvitrectomy. AB - This study was undertaken to determine the concentration of malondialdehyde, an end product of lipoperoxidation, in lens and retinal tissue postvitrectomy associated with oral administration of antioxidant flavonoids cyaninosides chloride and chromocarb diethylamine salt or N -acetylcysteine. Fifty adult pigmented rabbits were divided into five groups: (1) controls (normal eyes, malondialdehyde concentration in lens and retina); (2) vitrectomy with BSS Plus (malondialdehyde level measured 2hr after vitrectomy); (3) vitrectomy with BSS Plus and pretreatment with oral cyaninosides chloride 100mg kg day(-1)for 3 weeks (malondialdehyde level measured 2hr after surgery); (4) vitrectomy with BSS Plus and pretreatment with oral chromocarb diethylamine salt 100 mg kg day(-1)for 3 weeks (malondialdehyde level measured 2hr after surgery); and (5) vitrectomy with BSS Plus and pretreatment with oral N -acetylcysteine 200 mg kg day(-1)for 3 weeks (malondialdehyde level measured 2hr after surgery). Lens and retina samples were used to determine malondialdehyde levels using ion-pairing high performance liquid chromatography. Statistical analysis was done using analysis of variance (P<0.05). The content of malondialdehyde in the normal lens was 0.036 +/- 0.017 microg g(-1); in the vitrectomized groups the malondialdehyde concentrations were as follows: (2) 0.027 +/- 0.013 microg g(-1); (3) under detection limit (detection limit=1.75x e-3 microg g(-1)); (4) under detection limit; and (5) 0.020 +/- 0.006 microg g(-1). The results showed that the malondialdehyde concentration in the normal retina was 1.160 +/- 0.361 microg g(-1), while in the vitrectomized groups with or without pretreatment (cyaninosides chloride, chromocarb diethylamine salt, and N -acetylcysteine) the malondialdehyde levels were 2.091 +/- 0.982 microg g(-1), 0.069 +/- 0.024 microg g(-1), 0.082 +/- 0.027 microg g(-1), and 0.215 +/- 0.134 microg/g(-1), respectively, all significantly different from the normal eyes (P<0.05). Vitrectomy induced increased malondialdehyde levels in the retina. Oral flavonoids are an effective protective therapy for surgically induced lipoperoxidation, especially in the retina. PMID- 11878817 TI - Comparison between isopropyl unoprostone and latanoprost by prostaglandin E(2)induction, affinity to prostaglandin transporter, and intraocular metabolism. AB - The pharmacological differences between isopropyl unoprostone (referred to as unoprostone) and latanoprost, concerning their induction of endogenous prostaglandin E(2)(PGE(2)) and affinity to a human prostaglandin transporter (PGT), were investigated. Freshly dissected bovine iris tissues were incubated with major intraocular metabolites of unoprostone, M1 and M2, acid of latanoprost, or PGF(2 alpha), and PGE(2)induction was measured. Affinities of M1, M2, latanoprost, acid of latanoprost, and PGF(2 alpha)to PGT molecule were measured using PGT-cDNA transfected HeLa cells by an isotopic influx assay.(3)H unoprostone was incubated with freshly prepared serum, aqueous humor, or frozen stored fetal bovine serum (FBS), and the radioactivity of supernatants was measured to investigate their metabolism of(3)H-unoprostone.M2, acid of latanoprost, and PGF(2 alpha)significantly increased a release of PGE(2)compared with the control. 10 microM indomethacin completely inhibited PGE(2)induction by acid of latanoprost and PGF(2 alpha), while 100 microM indomethacin was required to inhibit PGE(2)induction completely by M1 and M2. Unoprostone, M1, M2, and latanoprost showed little affinity to PGT, while acid of latanoprost had an affinity to PGT. Freshly prepared serum and aqueous humor metabolized unoprostone, but frozen stored FBS did not. The release of endogenous PGE(2)may play an important role of action by means of PG analogs, and differences in indomethacin-related inhibition of PGE(2)release and in affinities to PGT may in part cause their different actions. PMID- 11878818 TI - Transscleral Coulomb-controlled iontophoresis of methylprednisolone into the rabbit eye: influence of duration of treatment, current intensity and drug concentration on ocular tissue and fluid levels. AB - The major problems associated with the use of corticosteroids for the treatment of ocular diseases are their poor intraocular penetration to the posterior segment when administered locally and their secondary side effects when given systemically. To circumvent these problems more efficient methods and techniques of local delivery are being developed. The purposes of this study were: (1) to investigate the pharmacokinetics of intraocular penetration of hemisuccinate methyl prednisolone (HMP) after its delivery using the transscleral Coulomb controlled iontophoresis (CCI) system applied to the eye or after intravenous (i.v.) injection in the rabbit, (2) to test the safety of the CCI system for the treated eyes and (3) to compare the pharmacokinetic profiles of HMP intraocular distribution after CCI delivery to i.v. injection. For each parameter evaluated, six rabbit eyes were used. For the CCI system, two concentrations of HMP (62.5 and 150mg ml(-1)), various intensities of current and duration of treatment were analyzed. In rabbits serving as controls the HMP was infused in the CCI device but without applied electric current. For the i.v. delivery, HMP at 10mg kg(-1)as a 62.5mg ml(-1)solution was used. The rabbits were observed clinically for evidence of ocular toxicity. At various time points after the administration of drug, rabbits were killed and intraocular fluids and tissues were sampled for methylprednisolone (MP) concentrations by high pressure liquid chromatography (HPLC). Histology examinations were performed on six eyes of each group. Among groups that received CCI, the concentrations of MP increased in all ocular tissues and fluids in relation to the intensities of current used (0.4, 1.0 and 2.0mA/0.5cm(2)) and its duration (4 and 10min). Sustained and highest levels of MP were achieved in the choroid and the retina of rabbit eyes treated with the highest current and 10min duration of CCI. No clinical toxicity or histological lesions were observed following CCI. Negligible amounts of MP were found in ocular tissues in the CCI control group without application of current. Compared to i.v. administration, CCI achieved higher and more sustained tissue concentrations with negligible systemic absorption. These data demonstrate that high levels of MP can be safely achieved in intraocular tissues and fluids of the rabbit eye, using CCI. With this system, intraocular tissues levels of MP are higher than those achieved after i.v. injection. Furthermore, if needed, the drug levels achieved with CCI can be modulated as a function of current intensity and duration of treatment. CCI could therefore be used as an alternative method for the delivery of high levels of MP to the intraocular tissues of both the anterior and posterior segments. PMID- 11878819 TI - Initiation and characterization of keratinocyte cultures from biopsies of normal human conjunctiva. AB - The purpose of the present study was to establish and characterize serum-free epithelial cultures of normal human conjunctiva using fresh biopsy tissue. To this end, small pieces of normal conjunctiva were biopsied from patients undergoing routine cataract surgery. Fragments of the tissue were placed in explant culture in medium containing fetal bovine serum for approximately 1 week to promote epithelial cell outgrowth. Cultures were then passaged multiple times into serum-free medium. Cultures generated in this way were at least 95% keratinocytes and exhibited a typical epithelial morphology, which was dependent on the extracellular Ca(2+)concentration. Immunocytochemical staining revealed that E-cadherin, P-cadherin, and involucrin were present in the cultures, with distributions consistent with their in vivo localization patterns. Distribution of keratins 19, 3, and 4 in conjunctival epithelial cultures were also consistent with in vivo patterns and distinctly different from patterns observed in epithelial cultures similarly generated from cornea and foreskin. Hence, conjunctival keratinocyte cultures retain some tissue-specific markers and do not revert to a generic, culture phenotype. PMID- 11878820 TI - Differentiation-dependent increases in lactate dehydrogenase activity and isoenzyme expression in rabbit corneal epithelial cells. AB - Lactate dehydrogenase (LDH) and glucose-6-phosphate dehydrogenase (G-6-PDH) activities were studied during corneal epithelial growth and differentiation in cell culture. LDH and G-6-PDH activities increased up to 60 and 150-fold, respectively, when corneal epithelial cells constituted a differentiated four to five layered epithelium; these increases showed a similar time-course to the expression of K3 keratin. Immunostaining experiments showed that in growing colonies, LDH staining is stronger in those cells that are K3 positive; in contrast, in confluent four to five layered epithelia LDH and K3 were located in all cell layers, similar to the pattern found in frozen sections from rabbit central cornea. During growth and differentiation, the LDH isoenzyme set from corneal epithelial cells did not change; and it was different from those observed in cultured conjunctival, esophageal and epidermal cells. The augment in LDH activity was due to a 25-fold increase in the LDH-H mRNA and a 12-fold augment in LDH-M mRNA. A computer-assisted search led to identify AP2 and Sp1 binding sites in the LDH and G-6-PDH promoters, suggesting that their expression might share common regulatory mechanisms with the regulation of the differentiation-linked keratins. It is proposed that LDH may be an early marker of corneal epithelial differentiation, and its isozyme pattern could be distinctive from other epithelial cell lineages. PMID- 11878821 TI - Metallothionein protects retinal pigment epithelial cells against apoptosis and oxidative stress. AB - The retina expresses metallothionein (MT) which has been reported to protect cells against oxidative stress and apoptosis. The types of MT expressed by human retinal cells were identified by laser capture microdissection and RT--PCR and it was found that MT-2a is expressed by retinal pigment epithelial (RPE) cells, photoreceptor cells, inner nuclear layer cells and ganglion cells while MT-1a is expressed by RPE cells and MT-3 by cells of the neural retina. MT is induced in cultured human RPE cells under stress conditions such as the presence of glucocorticoids, interleukin-1/TNF alpha, oxygen and TGF beta 1. Cultured human D407 RPE cells were transfected with plasmids that allowed the expression of MT to be controlled via the tet operator protein by the level of tetracycline in the medium. These experiments showed that elevation of MT levels by transfection of RPE cells protects them against toxic levels of cadmium, heme- and iron-induced oxidation and UV light-induced apoptosis. PMID- 11878822 TI - A clinical study of the human lens with a dynamic light scattering device. AB - A study was conducted to determine the potential usefulness and repeatability of a new dynamic light scattering (DLS) device for clinical studies of the human lens and early cataract. Studies using the cold cataract model showed this new device to be more sensitive than the Scheimpflug cataract imaging system in detecting the earliest cataractous changes. A miniaturized clinical DLS device developed by NASA using fiber optic probes was mounted on a Keratoscope (Optikon 2000), which has a 3-dimensional aiming system for accurate repeated sampling of the same area of the lens. A test/retest study was then conducted on the nuclear region of the lenses of 12 normal eyes. After a full, dilated eye examination, DLS data were obtained using the new device on the same eyes twice, 30-60 min apart. Particle size distributions and mean log particle size data were obtained. The mean percent differences between the larger and smaller of the test--retest pairs was 6.4% (range 0.05--10.8%); the between-test S.D. was 0.116. Actual numerical margin of error was +/- 0.023. In addition, the mean coefficient of variation was 4.2% (range 0.3--7.3%). A useful clinical end point obtained from data produced by the device was the mean log particle size. These results suggest that the DLS will be useful in the detection and study of the beginning and earliest stages of cataract formation in humans. PMID- 11878823 TI - Pirenzepine affects scleral metabolic changes in myopia through a non-toxic mechanism. AB - Whilst the precise mechanism regulating ocular growth is unknown, it has been shown that various pharmacological agents, including the muscarinic receptor antagonists, atropine and pirenzepine, are effective at preventing the development of myopia. A recent study, which demonstrated that muscarinic antagonists reduce the synthesis of glycosaminoglycans and DNA in chick sclera in vitro, led to the suggestion that such drugs may act directly on the sclera, possibly through a toxic mechanism. Accepted markers of scleral metabolism and cell viability were used in conjunction with a non-invasive, physiological method of ocular growth regulation to determine whether the selective muscarinic antagonist pirenzepine inhibits the development of myopia via toxicity to the sclera. Chicks were monocularly deprived (MD) of pattern vision and given daily intravitreal injections of either pirenzepine (700 microg) or saline vehicle into the deprived eye over 5 days. Unoccluded animals also received intravitreal injections of either pirenzepine or saline into one eye (n=6, all groups). The contralateral eye of all animals was left untreated for comparison. Optical and ocular biometric measures were collected on the final experimental day. Following in vivo delivery of [(35)S] labelled sulphate, levels of sulphate incorporation into scleral glycosaminoglycans were measured in proteinase K digests following selective precipitation with alcian blue dye. The DNA content was also assessed through luminescence spectrometry after binding to Hoechst 33258 dye. To allow comparison with an accepted non-invasive, physiological method of ocular growth regulation, myopia was prevented in additional groups of MD animals by allowing 3hr of unoccluded vision each day, over 5 days, before levels of sulphate incorporation were measured. Scleral DNA content, a marker of cell viability, was not significantly altered between treated and control eyes in any injected group. Relative levels of sulphate incorporation (% difference between treated and contralateral control eyes) were significantly lower in the cartilaginous sclera of pirenzepine-MD animals, compared to saline-MD controls (+35.9 +/- 10.1% vs +121.2 +/- 28.6%, P<0.05), after 2hr of incorporation. However, after 6hr incorporation, there was no significant difference in sulphate incorporation in the cartilaginous sclera between the two groups (+87.2 +/- 33.1% vs +111.0 +/- 14.4%, P=0.53). No significant change was found in the levels of glycosaminoglycan synthesis in the fibrous sclera of any pirenzepine treated group, when compared to the appropriate saline control. Relative patterns of sulphate incorporation, between treatment and control groups, were essentially identical at both time points examined, regardless of whether myopia was prevented through pirenzepine injection or periods of unoccluded vision. The present study shows that, at a dose of pirenzepine sufficient to prevent experimentally-induced axial myopia, glycosaminoglycan synthesis in the cartilaginous sclera was significantly reduced for a transient period following the injection. These pirenzepine-induced reductions in glycosaminoglycan synthesis were not caused by direct drug toxicity to scleral cells as these changes were reversible and no significant reduction in DNA content was observed in pirenzepine treated eyes. Similar patterns of scleral glycosaminoglycan synthesis changes were found following the provision of brief periods of unoccluded vision further demonstrating that pirenzepine is effective in myopia prevention via a non-toxic mechanism. Consequently, the prevention of myopia development in chicks, with either pirenzepine or brief periods of unoccluded vision, is associated with the transient modulation of scleral glycosaminoglycan synthesis in the cartilaginous sclera. PMID- 11878824 TI - Effect of H(2)O(2)on human lens epithelial cells and the possible mechanism for oxidative damage repair by thioltransferase. AB - Human lens epithelial (HLE) B3 cells were used to study the oxidative damage and cellular repair with respect to the redox homeostasis, the oxidative defense enzymes and the glucose metabolic pathway. The effect of oxidative stress on cell growth was initially analyzed by culturing the cells with a bolus amount (0.02- 0.1m M) of hydrogen peroxide (H(2)O(2)) in minimal essential medium (MEM) containing 20% fetal bovine serum (FBS) for 1 week. Concentration of H(2)O(2)greater than 0.03m M showed progressive inhibition of cell growth. However, the cells were also shown to tolerate H(2)O(2)concentrations up to 0.5m M by detoxifying the exogenous oxidant within 3hr without any detectable DNA damage. Therefore, this short-term H(2)O(2)exposure model was chosen to study the effect of oxidative stress on the cellular redox homeostasis. HLE B3 cells were first grown to confluence in MEM with 20% FBS. Approximately 1.6 million cells were gradually weaned off serum by subculturing in 2% FBS overnight, followed by serum-free medium for 30 min before subjecting to a bolus of 0.1m M H(2)O(2)for up to 180 min. These cells were used for biochemical analysis, which included H(2)O(2)detoxification (H(2)O(2)in the medium), glutathione (GSH) level and lactate production. Activity measurements were conducted on the oxidation defense enzymes: glutathione-S-transferase (GST), glutathione reductase (GR) and glutathione peroxidase (GPx); the dethiolating enzyme, thioltransferase (TTase); and a key glycolytic enzyme, glyceraldehyde-3-phosphate dehydrogenase (G-3PD). While the B3 cells were shown to tolerate and detoxify 0.1m M H(2)O(2)within 60 min, the GSH pool was transiently depleted in the first 60 min before fully recovered. GPx suffered more than 80% loss in activity and was unable to recover fully. GST showed slight inactivation but neither GR nor TTase was affected. G 3PD was inactivated to < 50% within 15 min of oxidative stress and was reactivated gradually to 80% of normal at the end of 180 min, concurrent with the transient loss of lactate production in the same cells. The reactivation of G-3PD was both temperature- and GSH-dependent, occurring only at physiological temperature and failing to reactivate when the intracellular GSH pool was depleted by BCNU (GR inhibitor) pretreatment. The inactivated cellular G-3PD in the cell extract could be partially reactivated by DTT (6m M) or by recombinant human lens thioltransferase (RHLT) but not by GSH (1m M), GR or GST. HLE cells cultured in the presence of L-(35)S-cystine and cycloheximide displayed an extra radiolabelled protein band on the autoradiograph in the H(2)O(2)treated cells. The labelled band was positively reacted with G-3PD antibody and could be removed by RHLT, indicating that S-thiolation of G-3PD occurred. The H(2)O(2)pre-exposed cells also transiently accumulated proteins modified by thiolation, including protein-S-S-glutathione (PSSG) and protein-S-S-cysteine (PSSC). It can be concluded that HLE could endure up to 0.1m M of H(2)O(2)oxidative stress since the cell could be protected by its effective repair systems, including dethiolating the inactivated key SH-sensitive enzymes. TTase may play a role in this. One of the mechanisms may be through preserving glucose metabolism and supplying ATP needed for maintaining cell viability. PMID- 11878825 TI - Co-localization of junction-associated proteins of the human blood--aqueous barrier: occludin, ZO-1 and F-actin. AB - Recent studies in mouse and rabbit eyes have begun to identify the molecular constituents of tight and adherens junctions that represent the structural equivalent of the blood--aqueous barrier (BAB). These species are commonly used as experimental models to examine the pathobiology of anterior uveitis, an inflammatory condition in which the junctions of the BAB are compromised. Because it was unclear whether major molecular elements of the junctions in these species were the same as those in humans, the goal of this study was to determine if the junction related proteins ZO-1 and occludin are present in normal human ciliary epithelium and iridial vascular endothelium. To determine their presence, sections of human anterior uvea were probed in 14 normal, human, eyebank eyes immunolabelled with antibodies to ZO-1, and occludin, and confocal microscopy was used to examine them. Phalloidin staining for F-actin was also assessed. ZO-1 and occludin were both localized along the apico-lateral surfaces of the non pigmented ciliary epithelium and the interendothelial clefts of iris blood vessels. In both locations, the distribution of occludin was more focal than seen for ZO-1. ZO-1 was also found along the apical surfaces between the pigmented and non-pigmented ciliary epithelial cell layers. The distribution of these proteins supports the notion that occludin is more specifically associated with tight junctions than is ZO-1 in the normal human BAB. No change in this distribution was found with increasing age. These data are consistent with findings reported previously in rabbit ciliary epithelium and iridial vascular endothelium, indicating the relevance of experimental induced uveitis studies in rabbit, as a model of BAB breakdown in human uveitis. PMID- 11878826 TI - Inhibition of murine corneal allograft rejection by treatment with antibodies to CD80 and CD86. AB - The purpose of this study was to investigate the role of CD80 and CD86 costimulatory molecules in corneal allograft rejection. Anti-CD80 and anti-CD86 monoclonal antibodies (mAbs) were administered after orthotopic corneal allograft transplantation. Graft rejection was observed by biomicroscopy. Population and localization of CD80(+)and CD86(+)cells in the cornea, cervical lymph nodes, and spleen were examined by flow cytometry and immunohistochemistry. The combined use of anti-CD80 and anti-CD86 mAbs was effective in prolonging corneal allograft survival. In the untreated mice bearing rejected graft, many CD86(+)and CD80(+)cells were found around the host-graft junctional area in the cornea, and CD86(high)cells were found in the cervical lymph node and spleen. In contrast, few CD86(+)or CD80(+)cells were observed in the cornea, cervical lymph node, and spleen from the mice treated with anti-CD80/CD86 mAbs. These results demonstrated a significant role of CD80 and CD86 costimulatory molecules in corneal allograft rejection. PMID- 11878827 TI - Drusen are Cold Spots for Proteolysis: Expression of Matrix Metalloproteinases and Their Tissue Inhibitor Proteins in Age-related Macular Degeneration. AB - Drusen are abnormal extracellular matrix deposits characteristic of age-related macular degeneration (AMD), a leading cause of blindness in the aging human population. The mechanisms underlying drusen formation are not well characterized. The purpose of this study was to examine the expression of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) in drusen, and in the surrounding cells and tissue. To assess the extent of MMP and TIMP expression by retinal pigment epithelial (RPE) cells, cDNA arrays were screened with probes generated from cultured human RPE cells. The distribution of MMP-1, -2 and -3 and TIMP-1, -2, -3 and -4 was determined using immunohistochemistry in human RPE choroid from donor eyes with and without a clinical history of AMD. Gelatinase activity was assessed in unfixed frozen sections using in situ zymography. In cultured RPE cells, expression of 10 MMP and all four known TIMP mRNAs was detected. MMP immunoreactivity was widespread in the RPE choroid, but was absent from the interior of drusen. TIMP-3, but not other TIMPs, was detected in the drusen interior. Likewise, metal ion dependent gelatinase activity could be detected in RPE choroid, but not in drusen. These results show that, while metalloproteinase activity is widespread throughout the RPE choroid, drusen are cold spots for proteolysis. The data lead to the speculation that high TIMP-3 concentrations within drusen could inhibit MMPs and as a result slow the proteolytic degradation of these deposits. PMID- 11878828 TI - Microablation of choroidal tissue from Bruch's membrane using a 193nm excimer laser. PMID- 11878829 TI - A mixed ectoparasite--microparasite model for bat-transmitted rabies. AB - This paper considers the transmission of rabies to domestic livestock by vampire bats. Vampire bats act as ectoparasites on cattle both by ingesting a small amount of blood every night and by prolonging bleeding by the action of anticoagulant substances in their saliva. In addition to this parasitic action bats may also transmit rabies, inflicting important losses on affected herds by the inevitable mortality due to the infection. We modeled this complex interaction and we also demonstrate that bat control measures are more effective in reducing rabies prevalence and mortality by rabies than cattle vaccination. PMID- 11878830 TI - Spatially structured metapopulation models: global and local assessment of metapopulation capacity. AB - We model metapopulation dynamics in finite networks of discrete habitat patches with given areas and spatial locations. We define and analyze two simple and ecologically intuitive measures of the capacity of the habitat patch network to support a viable metapopulation. Metapopulation persistence capacity lambda(M) defines the threshold condition for long-term metapopulation persistence as lambda(M)>delta, where delta is defined by the extinction and colonization rate parameters of the focal species. Metapopulation invasion capacity lambda(I) sets the condition for successful invasion of an empty network from one small local population as lambda(I)>delta. The metapopulation capacities lambda(M) and lambda(I) are defined as the leading eigenvalue or a comparable quantity of an appropriate "landscape" matrix. Based on these definitions, we present a classification of a very general class of deterministic, continuous-time and discrete-time metapopulation models. Two specific models are analyzed in greater detail: a spatially realistic version of the continuous-time Levins model and the discrete-time incidence function model with propagule size-dependent colonization rate and a rescue effect. In both models we assume that the extinction rate increases with decreasing patch area and that the colonization rate increases with patch connectivity. In the spatially realistic Levins model, the two types of metapopulation capacities coincide, whereas the incidence function model possesses a strong Allee effect characterized by lambda(I)=0. For these two models, we show that the metapopulation capacities can be considered as simple sums of contributions from individual habitat patches, given by the elements of the leading eigenvector or comparable quantities. We may therefore assess the significance of particular habitat patches, including new patches that might be added to the network, for the metapopulation capacities of the network as a whole. We derive useful approximations for both the threshold conditions and the equilibrium states in the two models. The metapopulation capacities and the measures of the dynamic significance of particular patches can be calculated for real patch networks for applications in metapopulation ecology, landscape ecology, and conservation biology. PMID- 11878831 TI - The use of Markovian metapopulation models: a comparison of three methods reducing the dimensionality of transition matrices. AB - The use of Markovian models is an established way for deriving the complete distribution of the size of a population and the probability of extinction. However, computationally impractical transition matrices frequently result if this mathematical approach is applied to natural populations. Binning, or aggregating population sizes, has been used to permit a reduction in the dimensionality of matrices. Here, we present three deterministic binning methods and study the errors due to binning for a metapopulation model. Our results indicate that estimation errors of the investigated methods are not consistent and one cannot make generalizations about the quality of a method. For some compared output variables of populations studied, binning methods that caused a strong reduction in dimensionality of matrices resulted in better estimations than methods that produced a weaker reduction. The main problem with deterministic binning methods is that they do not properly take into account the stochastic population process itself. Straightforward usage of binning methods may lead to substantial errors in population-dynamical predictions. PMID- 11878832 TI - Evolution of altruism in stepping-stone populations with overlapping generations. AB - We study the evolution of altruism in one- and two-dimensional stepping-stone populations with discrete overlapping generations. We find that increasing survival probability facilitates the evolution of altruism, in agreement with recent results for a patch-structured population. We allow the altruistic behaviour to affect either fecundity or survival probability. In the first case, altruism is favoured compared to a randomly interacting population, but in the second case, altruism is less likely to evolve. We consider the iterated prisoner's dilemma as a description of an altruistic interaction and compare our results with recent simulations of lattice populations. PMID- 11878833 TI - Selection and segregation distortion in a sex-differentiated population. AB - We extend the classical model for selection at an autosomal locus in a sex differentiated population to include segregation distortion. The equations remain the same, but the fitness parameters are interpreted differently and refer to alleles instead of genotypes. We derive conditions for internal and external stability of the equilibria, i.e., stability with respect to perturbations of alleles that are already present at equilibrium and stability with respect to invasion attempts by newly arising alleles. We show that, in a sex-differentiated population, external stability of an equilibrium can be judged on the basis of Shaw--Mohler criteria. Throughout, we compare the situation in populations with and without sex differentiation. Interestingly, internal stability is more difficult to achieve in a population without sex differentiation than in a population in which selection and segregation distortion are restricted to one sex. In a companion paper we show how the general results of the present paper can lead to new insights into specific systems such as the t complex of the house mouse. PMID- 11878834 TI - Competition at the mouse t complex: rare alleles are inherently favored. AB - We investigate the competition between alleles at a segregation distorter locus. The focus is on the invasion prospects of rare mutant distorter alleles in a population in which a wildtype and a resident distorter allele are present. The parameters are chosen to reflect the situation at the t complex of the house mouse, one of the best-studied examples of segregation distortion. By analyzing the invasion chances of rare alleles, we provide an analytical justification of earlier simulation results. We show that a new distorter allele can successfully invade even if it is inferior both at the gamete and at the individual level. In fact, newly arising distorter alleles have an inherent rareness advantage if their negative fitness consequences are restricted to homozygous condition. Likewise, rare mutant wildtype alleles may often invade even if their viability or fertility is reduced. As a consequence, the competition between alleles at a segregation distorter locus should lead to a high degree of polymorphism. We discuss the implications of this conclusion for the t complex of the house mouse and for the evolutionary stability of "honest" Mendelian segregation. PMID- 11878872 TI - Coreceptor usage of sequential isolates from cynomolgus monkeys experimentally Infected with simian immunodeficiency virus (SIVsm). AB - Sequential isolates from eight cynomolgus monkeys experimentally infected with simian immunodeficiency virus (SIVsm, of sooty mangabey origin) were tested for coreceptor use in the human osteosarcoma indicator cell line, GHOST(3), expressing CD4 and one or another of the chemokine receptors CCR3, CCR5, CXCR4, BOB, or the orphan receptor Bonzo. The indicator cell line carries the human immunodeficiency virus type 2 long terminal repeat-driven green fluorescence protein gene that becomes activated upon infection with HIV or SIV and fluorescence can be quantitated by flow cytometric analysis. The methodological details are described in the accompanying paper (Vodros et al., 2001, Virology 290, in press). All SIVsm inoculum viruses and reisolates used CCR5 with a high level of efficiency. CCR5 use was stable over time. BOB and Bonzo use was less efficient than CCR5 use and, in particular, late isolates obtained at the time of immunodeficiency varied greatly in their coreceptor use and often could not establish a productive infection in BOB- or Bonzo-expressing cells. Unexpectedly, early reisolates obtained 12 days postinfection could infect the entire GHOST(3) panel including the parental cells. In one case this was due to use of CXCR4, either transfected or endogenously expressed on the GHOST(3) cells. Our results demonstrate the complex coreceptor use of SIVsm isolates. Moreover, they focus attention on the initial phase of virus replication when the availability of target cells may govern the replication pattern of the virus. PMID- 11878873 TI - Characterization of a baculovirus-encoded protein that is associated with infected-cell membranes and budded virions. AB - Two types of envelope fusion proteins have been identified in lepidopteran baculoviruses. GP64 is found in Autographa californica multinucleocapsid nucleopolyhedrovirus, Orgyia pseudotsugata multinucleocapsid nucleopolyhedrovirus (OpMNPV), and other relatively closely related viruses, while Lymantria dispar multinucleocapsid nucleopolyhedrovirus (LdMNPV), which lacks GP64, utilizes LD130 as its envelope fusion protein. Homologs of ld130 have since been found not only in all the sequenced gp64-minus virus genomes, but also in the genomes of gp64 containing viruses. In addition, they are evolutionarily related to the envelope proteins of certain insect retroviruses. In this report, the characterization of a LD130 homolog (OP21) from OpMNPV, which also contains gp64, is described. Western blot analysis of extracts of OpMNPV-infected Lymantria dispar cells, using antibodies generated against OP21, identified an infected cell-specific doublet of 85 and 89 kDa. These bands were first observed at about 6 h p.i. and were present at all later time points. Such analyses also demonstrated that OP21 was associated with budded virions. Tunicamycin treatment of OpMNPV-infected cells indicated that OP21 is N-glycosylated. Studies employing NP-40 to remove the envelope from budded virions indicated that the majority of OP21 remained associated with the nucleocapsid fraction, whereas all GP64 was removed. Confocal immunofluorescence microscopy showed that OP21 and GP64 have a similar pattern of distribution on the membrane of cells infected with OpMNPV. Immunoelectron microscopy of budded virions also showed similar patterns of localization for both OP21 and GP64. PMID- 11878871 TI - Quantitative evaluation of HIV-1 coreceptor use in the GHOST3 cell assay. AB - The utility of the GHOST(3) cell assay has been evaluated for testing coreceptor use of primary human immunodeficiency virus type 1 (HIV-1) isolates. GHOST(3) cells were derived from the human osteosarcoma cell line, HOS, and have been engineered to stably express CD4 and one or another of the chemokine receptors CCR3, CCR5, CXCR4, Bonzo, or the orphan receptor BOB. The indicator cell line carries the HIV-2 long terminal repeat-driven green fluorescence protein (GFP) gene, which becomes activated upon infection with HIV or simian immunodeficiency virus. Viral entry is followed by Tat activation of transcription and GFP becomes expressed. Infected cells can be detected 2 or 3 days after infection by simple fluorescence microscopic observation. This simplicity is the main advantage of the GHOST(3) cell system and makes it particularly suitable for screening of a large number of isolates. In addition, the efficiency of coreceptor use can be accurately quantitated with flow cytometric analysis. Here, we evaluated the coreceptor use of 59 primary HIV-1 isolates of different subtypes. PMID- 11878874 TI - Mutational analysis of the N-terminus of Moloney murine leukemia virus integrase. AB - The retroviral integrase (IN) carries out the integration of viral DNA into the host genome. The IN protein consists of three domains: the N-terminal HHCC motif, the catalytic core region, and the C-terminus. The Moloney murine leukemia virus (M-MuLV) IN encodes a unique 45-amino-acid domain N-terminal to the HHCC motif. The function of the N-terminus of M-MuLV IN was studied through deletional and mutational analyses. The IN 1-105 domain was dissected into two halves expressing either the unique N-terminus or the HHCC domain. Although the parental IN 1-105 could functionally complement the core-C-terminus for integration reactions, neither half of the N-terminus was sufficient. Partial complementation of strand transfer, but not 3prime prime or minute processing, could be obtained through mixing the two halves. The dimerization of the M-MuLV N-terminus was dependent on the expression of the intact 1-105. Critical basic amino acids within the HHCC domain which are required for 3' processing and strand transfer reactions were identified through alanine mutagenesis. Loss of in vitro strand transfer activity correlated with loss of viral titer in vivo for this cluster of basic amino acids within the HHCC domain. PMID- 11878875 TI - LMP2A survival and developmental signals are transmitted through Btk-dependent and Btk-independent pathways. AB - The latent membrane protein 2A (LMP2A) of Epstein--Barr virus (EBV) has been implicated in controlling viral latency due to the ability of LMP2A to block B cell antigen receptor (BCR) signaling in vitro and to alter B cell development and enhance B cell survival in vivo. These LMP2A functions require interactions with the protein tyrosine kinases Syk and Lyn. However, a role for the Bruton's tyrosine kinase (Btk) has not been investigated for these LMP2A functions. To investigate whether Btk is important for LMP2A developmental and survival signals in vivo, LMP2A transgenic animals were mated to Btk deficient (Btk(-/-)) mice. Unlike LMP2A(+), Btk(+/+) transgenic littermate controls, LMP2A(+), Btk(-/-) animals do not generate immunoglobulin (Ig) receptorless B cells in the periphery and instead produce Ig(+) B cells similar to those in the Btk(-/-) mice. Interestingly, however, LMP2A(+), Btk(-/-) animals produce B cells at a vastly reduced level compared to Btk(-/-) littermates, indicating that LMP2A affects B cell development in the absence of Btk. In the RAG-1(-/-), Btk(-/-) double knockout background, LMP2A is still capable of enhancing the survival of Ig receptorless B cells. Use of Btk phosphopeptide-specific antibodies reveals that Btk is constitutively phosphorylated in LMP2A-expressing cell lines. These data indicate that LMP2A initiates both Btk-dependent and Btk-independent pathways, resulting in altered B cell development and enhanced B cell survival. PMID- 11878876 TI - Persistent infection of B lymphocytes by bovine respiratory syncytial virus. AB - Bovine respiratory syncytial virus (BRSV) is a major cause of respiratory disease in young cattle. Here we demonstrate BRSV persistence at low levels in tracheobronchial and mediastinal lymph nodes up to 71 days after the experimental infection of calves. Positive results were obtained on viral genomic RNA and messenger RNA coding for the nucleoprotein, glycoprotein (G), and fusion protein (F). G and F proteins were also detected in the pulmonary lymph nodes by immunohistochemistry. Double-staining experiments revealed that viral antigen was present in B-lymphocytes. Coculture experiments with the lymph node cells showed that the virus was still able to infect permissive target cells, even though no cytopathic effect was recorded. In vitro studies indicate that BRSV was still able to replicate in bovine B-lymphocyte cell lines 6 months after infection. These results may also be relevant to the understanding not only of the epidemiology and the peculiarities of the immune response of BRSV infections but also of human respiratory syncytial virus infections. PMID- 11878877 TI - Analysis of early region 1 of porcine adenovirus type 3. AB - To identify the proteins encoded by the porcine adenovirus 3 (PAV-3) E1 region, rabbit antisera were prepared using a bacterial fusion protein encoding E1A, E1B(small), or E1B(large) protein. Sera against E1A, E1B(small), and E1B(large) immunoprecipitated a protein of 35, 23, and 53 kDa, respectively, in in vitro translated and transcribed mRNA and PAV-3 infected cells. To determine the role of E1 proteins in PAV-3 replication, we constructed vectors with a deletion(s) in the E1 region. Mutant PAV211, containing deletions in E1A and E3, grew to titers similar to wild-type in VIDO R1 cells (E1A complementing) but not in swine testicular (ST) cells. No early protein (E1B(small), DNA binding protein) expression could be detected in PAV211 infected ST cells by Western blots. Mutant PAV212, containing deletions in E1B(small) and E3, grew to wild-type titers in VIDO R1 or ST cells. These deletions were successfully rescued, resulting in recombinant PAV214, containing deletions in E1A, E1B(small), and E3. However, mutant PAV-3, containing a triple stop codon inserted in the E1B(large) coding sequence, could not be isolated. Next, we constructed a recombinant PAV216 by inserting the green fluorescent protein gene flanked by a promoter and a poly(A) in the E1A region of the PAV214 genome. Both PAV214 and PAV216 replicate as efficiently as wild-type in VIDO R1 cells. These results suggested that (a) E1A is essential for virus replication and is required for the activation of other PAV-3 early genes, (b) E1B(small) is not essential for replication of PAV-3, and (c) E1B(large) is essential for virus replication. Moreover, the PAV216 vector can be used for the expression of a transgene. PMID- 11878878 TI - A family of closely related bovine viral diarrhea virus recombinants identified in an animal suffering from mucosal disease: new insights into the development of a lethal disease in cattle. AB - Induction of lethal mucosal disease (MD) in cattle is linked to the generation of cytopathogenic (cp) bovine viral diarrhea virus (BVDV) in animals persistently infected with a noncytopathogenic BVDV. In most cases the cp variants are generated by recombination with cellular or viral sequences. BVDV was obtained from the serum of an MD animal and propagated in tissue culture without plaque purification. Analysis of cDNA clones established from RNA of these cells showed that apparently a variety of different viral RNAs were present. Seven of the cDNA clones contained a cellular sequence coding for light chain 3 (LC3) of microtubule-associated proteins 1A and 1B. This insertion had already been found in the cp virus JaCP obtained from the same animal and isolated by plaque purification. Analysis of further plaque-purified cp viruses showed that the diseased animal contained a family of closely related cp BVDV recombinants. A set of viruses with different duplications of viral sequences in their genomes and a variety of defective viral RNAs with deletions were found that all contained the LC3* insertion. For all the recombinants the 3' recombination sites and, in all but one case, also the 5' recombination sites between cellular and viral sequence were identical. Variation between the individual deduced genome structures resulted from different duplications or deletions of viral sequences located upstream of the cellular insertion. These results suggest that within the animal a primary recombinant with a genome containing the LC3* insertion was generated. In a trimming process a set of secondary virus recombinants was generated from this hypothetical primary recombined RNA. These secondary recombinants display genome structures that represent variations of the basic scheme already present in the primary recombinant. Apparently this trimming process that finally led to an outbreak of MD lasted a long time since recombined RNA with the basic genome structure of the cp viruses could be demonstrated in samples already taken a long time before outbreak of the disease. PMID- 11878879 TI - Amphotropic murine leukemia virus replication in human mammary epithelial cells and the formation of cytomegalovirus-promoter recombinants. AB - Amphotropic murine leukemia virus (MLV) can replicate in human cells and is a potential contaminant in vector preparations for human gene transfer studies. We have recently shown that replication of amphotropic MLV in specific human sarcoma and lymphoma lines is possible in the absence of the viral 75-bp transcription enhancer elements. Here, we have tested the replication of an amphotropic MLV, MLV-(MOA), and an enhancer-deficient mutant of this virus in human breast carcinoma-derived cell lines. The proviral expression plasmids use a cytomegalovirus (CMV) promoter for the initial transcription of virus RNA. We found that all cells analyzed are permissive for replication of MLV-(MOA). Enhancer-deficient virus is unable to replicate. However, in two lines the replication defect can be rescued by the spontaneous insertion of a CMV promoter and enhancer into the U3 region. This recombinant virus MLV-(RCMV) replicates with kinetics similar to that of MLV-(MOA) but is restricted to specific cell lines. The potential formation of RCMV recombinants during MLV vector preparation must be considered. PMID- 11878880 TI - Promoter strength in adenovirus transducing vectors: down-regulation of the adenovirus E1A promoter in 293 cells facilitates vector construction. AB - Most adenovirus transducing vectors have the cytomegalovirus major immediate early (CMV) or the Rous sarcoma virus long terminal repeat (RSV) promoter driving expression of the transgene. Both of these promoters are highly active in transfection and transduction assays in 293 cells, in which transducing vectors are constructed and grown, and in HeLa cells. The CMV promoter exhibits rapid activation while the RSV promoter exhibits a lag prior to the onset of viral DNA replication in transduction assays. While the use of very strong promoters facilitates expression of the transgene, high-level expression of certain gene products hinders virus construction and growth. For such genes, the use of the adenovirus type 5 E1A promoter offers advantages. The E1A promoter exhibits modest activity in HeLa cells after transfection or transduction, but very little activity in 293 cells, suggesting that the E1A promoter would permit construction and growth of vectors encoding deleterious gene products that could not be constructed with the CMV and RSV promoters. This idea was tested through attempts to construct viruses encoding the immunoglobulin loop 6 and transmembrane regions of the prostaglandin F2alpha receptor regulatory protein (FPRP), a product that inhibits adenovirus vector construction for reasons that are not clear. Only the E1A promoter permitted construction and growth of the transducing vector encoding the fragment of FPRP. PMID- 11878881 TI - Functional analysis of proteins involved in movement of the monopartite begomovirus, Tomato yellow leaf curl virus. AB - The functional properties of proteins [capsid protein (CP), V1, and C4] potentially involved with movement of the monopartite begomovirus, Tomato yellow leaf curl virus (TYLCV), were investigated using microinjection of Escherichia coli expressed proteins and transient expression of GFP fusion proteins. The TYLCV CP localized to the nucleus and nucleolus and acted as a nuclear shuttle, facilitating import and export of DNA. Thus, the CP serves as the functional homolog of the bipartite begomovirus BV1. The TYLCV V1 localized around the nucleus and at the cell periphery and colocalized with the endoplasmic reticulum, whereas C4 was localized to the cell periphery. Together, these patterns of localization were similar to that of the bipartite begomovirus BC1, known to mediate cell-to-cell movement. However, in contrast to BC1, V1 and C4, alone or in combination, had a limited capacity to move and mediate macromolecular trafficking through mesophyll or epidermal plasmodesmata. Immunolocalization and in situ PCR experiments, conducted with tomato plants at three stages of development, established that TYLCV infection was limited to phloem cells of shoot apical, leaf, stem, and floral tissues. Thus, the V1 and/or C4 may be analogs of the bipartite begomovirus BC1 that have evolved to mediate TYLCV movement within phloem tissue. PMID- 11878882 TI - Complete genome analysis of the mandarin fish infectious spleen and kidney necrosis iridovirus. AB - The nucleotide sequence of the infectious spleen and kidney necrosis virus (ISKNV) genome was determined and found to comprise 111,362 bp with a G+C content of 54.78%. It contained 124 potential open reading frames (ORFs) with coding capacities ranging from 40 to 1208 amino acids. The analysis of the amino acid sequences deduced from the individual ORFs revealed that 35 of the 124 potential gene products of ISKNV show significant homology to functionally characterized proteins of other species. Some of the putative gene products of ISKNV showed significant homologies to proteins in the GenBank/EMBL/DDBJ databases including enzymes and structural proteins involved in virus replication, transcription, protein modification, and virus-host interaction. In addition, one major repeated sequence showing significant homology to the Red Sea bream iridovirus (RSIV) genome was identified. Based on the information obtained from biological properties (including histopathology, tissue tropisms, natural host range, and geographic distribution), physiochemical and physical properties, and genome analysis, we suggest that ISKNV, RSIV, sea bass iridovirus, grouper iridovirus, and African lampeye iridovirus may belong to a new genus of the Iridoviridae family and are tentatively referred to as cell hypertrophy iridoviruses. PMID- 11878883 TI - Inhibition of adenovirus cytotoxicity, replication, and E2a gene expression by adeno-associated virus. AB - Adeno-associated virus (AAV) and the other parvoviruses have long been known to inhibit proliferation of nonpermissive cells. The mechanism of this inhibition is not thoroughly understood. To learn how AAV interacts with host cells, we have begun an investigation into AAV's relationship with adenovirus (Ad), AAV's most efficient helper virus. AAV, but not UV-inactivated AAV, delayed Ad-induced cytotoxicity and inhibited Ad E2a gene expression. AAV, but not UV-inactivated AAV or a recombinant AAV vector, inhibited Ad DNA replication. To determine whether AAV or its replication (Rep) proteins alter Ad early gene expression, we measured steady state E2a mRNA levels in AAV and Ad coinfected cultures and in a cell line (Neo6) that inducibly expresses the Rep proteins. AAV, but not UV-AAV, and Rep expression resulted in diminution of E2a protein and mRNA levels. To determine whether the AAV Rep proteins directly affect the individual Ad early promoters, we constructed luciferase reporter plasmids containing each of the five early promoters. Cotransfection of Ad-luciferase and an AAV rep gene expressing plasmid in HeLa cells demonstrated that Rep78 repressed the E1a, E2a, and E4 promoters but trans-activated the E1b and E3 promoters. In the presence of a cotransfected E1a-expressing plasmid, Rep78 repressed expression from all five promoters. These results indicate that Rep may have different effects on the Ad early promoters dependent upon the presence of the E1a trans-activating protein. PMID- 11878884 TI - Rep and Rep' protein of porcine circovirus type 1 bind to the origin of replication in vitro. AB - Genome replication of Porcine circovirus type 1 (PCV1) relies upon expression of the full-length protein Rep and a spliced isoform (Rep'), and the presence of a 111-bp genomic fragment comprising the origin of replication. Using an electrophoretic mobility shift assay (EMSA), the capability of both Rep proteins to bind to partial fragments of the origin of replication of PCV1 was investigated in vitro. Both proteins formed complexes with double-stranded DNA origin fragments containing a stem-loop structure with a conserved nonamer and four hexamer repeats (5'-CGGCAG; H1 to H4). Use of truncated EMSA substrates identified minimal binding sites (MBS) for Rep and Rep' protein: The Rep binding site was mapped to the right leg of the stem-loop and the two inner hexamer repeats H1/H2, while binding of Rep' required only the presence of two hexamer repeats. Two differentially retarded complexes were observed with Rep protein, which presumably result from alternative binding to the MBS or to H3/4. PMID- 11878885 TI - Involvement of RNA2-encoded proteins in the specific transmission of Grapevine fanleaf virus by its nematode vector Xiphinema index. AB - The nepovirus Grapevine fanleaf virus (GFLV) is specifically transmitted by the nematode Xiphinema index. To identify the RNA2-encoded proteins involved in X. index-mediated spread of GFLV, chimeric RNA2 constructs were engineered by replacing the 2A, 2B(MP), and/or 2C(CP) sequences of GFLV with their counterparts in Arabis mosaic virus (ArMV), a closely related nepovirus which is transmitted by Xiphinema diversicaudatum but not by X. index. Among the recombinant viruses obtained from transcripts of GFLV RNA1 and chimeric RNA2, only those which contained the 2C(CP) gene (504 aa) and 2B(MP) contiguous 9 C-terminal residues of GFLV were transmitted by X. index as efficiently as natural and synthetic wild type GFLV, regardless of the origin of the 2A and 2B(MP) genes. As expected, ArMV was not transmitted probably because it is not retained by X. index. These results indicate that the determinants responsible for the specific spread of GFLV by X. index are located within the 513 C-terminal residues of the polyprotein encoded by RNA2. PMID- 11878886 TI - Rotavirus cross-species pathogenicity: molecular characterization of a bovine rotavirus pathogenic for pigs. AB - Rotaviruses which cause disease in heterologous animal species have been reported but the molecular basis of cross-species infectivity and disease is not established. We report the molecular characterization of a cloned rotavirus, PP 1, which was originally obtained from cattle and which had been biologically characterized in vivo in two target animal species, gnotobiotic pigs and calves. In pigs, PP-1 caused severe clinical disease but in experimental calves it replicated subclinically. PP-1 was characterized as a G3 reassortant with a porcine VP4 and NSP4 but a bovine NSP1. The PP-1 VP4 had 96 to 97% deduced amino acid identity to P[7] porcine rotaviruses and P[7] specificity was confirmed with VP4-specific monoclonal antibodies. Sequence analysis of the PP-1 NSP1 showed 94 to 99.6% deduced amino acid identity to bovine rotaviruses but the NSP4 protein had 94 to 98% identity to the NSP4 genotype B porcine rotaviruses. G-typing PCR initially classified PP-1 as a G10 rotavirus but sequence analysis revealed 92 to 96% identity of the PP-1 VP7 with porcine, simian, and human G3 rotaviruses. These results, combined with the in vivo properties of PP-1 in the two target species, supported the concept that species-specific VP4 and NSP4, but not NSP1, are required to induce rotavirus disease, at least in calves and pigs. The results illustrate experimentally that rotaviruses circulating in one animal species can pose a risk to another by the emergence of a pathogenic reassortant rotavirus under appropriate conditions. PMID- 11878887 TI - A reassortant bunyavirus isolated from acute hemorrhagic fever cases in Kenya and Somalia. AB - In late 1997 and early 1998, a large outbreak of hemorrhagic fever occurred in East Africa. Clinical samples were collected in Kenya and southern Somalia, and 27 of 115 (23%) hemorrhagic fever patients tested showed evidence of acute infection with Rift Valley fever (RVF) virus as determined by IgM detection, virus isolation, detection of virus RNA by reverse transcription-polymerase chain reaction (RT-PCR), or immunohistochemistry. However, two patients (one from Kenya and the other from Somalia) whose illness met the hemorrhagic fever case definition yielded virus isolates that were not RVF. Electron microscopy suggested these two virus isolates were members of the family Bunyaviridae. RT PCR primers were designed to detect bunyavirus RNA in these samples. Regions of the S and L segments of the two isolates were successfully amplified, and their nucleotide sequences exhibited nearly complete identity with Bunyamwera virus, a mosquito-borne virus not previously associated with severe human disease. Unexpectedly, the virus M segment appeared to be reassorted, as the sequences detected exhibited 32-33% nucleotide and 28% amino acid differences relative to the corresponding M segment sequence of Bunyamwera virus. The association of this reassortant bunyavirus, proposed name Garissa virus, with severe disease is supported by the detection of the virus RNA in acute-phase sera taken from 12 additional hemorrhagic fever cases in the region. PMID- 11878888 TI - A novel lentivirus vector derived from apathogenic simian immunodeficiency virus. AB - The improvement of gene transfer efficiency in growth-arrested cells using human immunodeficiency virus type 1 (HIV-1)-derived vectors led to the development of vectors derived from other members of the lentivirus family. Here we report the generation of a lentiviral vector derived from the apathogenic molecular virus clone SIVagm3mc of the simian immunodeficiency virus from African green monkeys (Cercocebus pygerythrus). Upon pseudotyping with the G-protein of vesicular stomatitis virus (VSV-G), the SIVagm-derived vector was shown to transduce proliferating and growth-arrested mammalian cell lines, including human cells. After in vivo inoculation into the striatum of the adult rat brain, the vector was shown to transduce terminally differentiated neurons and oligodendrocytes as well as quiescent and reactive astrocytes. Moreover, SIVagm transfer vector mRNA was efficiently packaged by HIV-1 vector particles. Homologous [SIV(SIV)] vectors generated by using the SIVagm-derived envelope glycoproteins allowed selective gene transfer into human CD4(+)/CCR5(+) cells. Thus, the SIVagm3mc-derived vector is a useful alternative to HIV-1-derived lentiviral vectors in somatic gene therapy. PMID- 11878889 TI - Association of Vpu-binding protein with microtubules and Vpu-dependent redistribution of HIV-1 Gag protein. AB - The efficient exit of HIV-1 particles from cells requires the action of the viral encoded protein Vpu. Vpu-binding protein (Ubp) is a cellular protein that interacts with both Vpu and the major structural component of the viral capsid (Gag) and appears to affect the efficiency of particle exit. Elucidation of the function of Ubp and characterization of the spatial distribution of Ubp may provide information pertinent to understanding the role of Ubp in virus replication. To investigate the subcellular location of Ubp, and to see whether Vpu affects the intracellular distribution of Gag, we carried out immunofluorescence localization in conjunction with confocal microscopy. Based on this analysis Ubp is present in both the nucleus and the cytoplasm. In the cytoplasm, Ubp appeared to be associated with microtubules as evidenced by cofluorescence with tubulin in the absence and in the presence of colchicine. However, cytoskeletal isolation and detergent extraction of cells resulted in association of Ubp with the soluble fractions, indicating that Ubp is not in tight association with microtubules. Moreover, flotation gradient analysis demonstrated that Ubp is cytoplasmic and not stably associated with the plasma membrane. Interestingly, expression of Vpu in cells resulted in redistribution of both Ubp and Gag to a location near the periphery of the cell. The effect of Vpu on both Ubp and Gag protein has implications for Vpu-mediated particle exit from cells. PMID- 11878890 TI - Analysis of the phosphorylation status of Epstein-Barr virus LMP2A in epithelial cells. AB - LMP2A deletion and point mutants, with mutations in phosphotyrosine-containing protein-protein interaction motifs, were transiently expressed in 293 cells and their phosphorylation was examined in immune complex kinase assays as well as in vivo. In vitro LMP2A phosphorylation depended on tyrosine 112. In vivo, mutations of single tyrosines did not eliminate LMP2 phosphorylation, although mutation of the LMP2A ITAM decreased LMP2A phosphorylation. The relationship between LMP2A in vitro phosphorylation and that induced by cell-extracellular matrix (ECM) interactions was also investigated. While LMP2A was phosphorylated to higher levels in whole-cell extracts of stimulated cells, a difference in in vitro kinase assays with extracts from stimulated and unstimulated cells was not detected, indicating that the ECM-mediated regulation of LMP2A phosphorylation is lost in vitro. In the presence of LMP2A, several cellular proteins with molecular weights between 70 and 80 kDa were phosphorylated on tyrosine. This increase in cellular protein phosphorylation depended on the LMP2A ITAM motif and suggests that the ITAM may participate in signal-transduction events in epithelial cells. PMID- 11878891 TI - Evidence that the hypermutated M protein of a subacute sclerosing panencephalitis measles virus actively contributes to the chronic progressive CNS disease. AB - Subacute sclerosing panencephalitis (SSPE) is a progressive degenerative disease of the brain uniformly leading to death. Although caused by measles virus (MV), the virus recovered from patients with SSPE differs from wild-type MV; biologically SSPE virus is defective and its genome displays a variety of mutations among which biased replacements of many uridine by cytidine resides primarily in the matrix (M) gene. To address the question of whether the SSPE MVs with M mutations are passive in that they are not infectious, cannot spread within the CNS, and basically represent an end-stage result of a progressive infection or alternatively SSPE viruses are infectious, and their mutations enable them to persist and thereby cause a prolonged neurodegenerative disease, we utilized reverse genetics to generate an infectious virus in which the M gene of MV was replaced with the M gene of Biken strain SSPE MV and inoculated the recombinant virus into transgenic mice bearing the MV receptor. Our results indicate that despite biased hypermutations in the M gene, the virus is infectious in vivo and produces a protracted progressive infection with death occurring as long as 30 to 50 days after that caused by MV. In primary neuron cultures, the mutated M protein is not essential for MV replication, prevents colocalization of the viral N with membrane glycoproteins, and is associated with accumulation of nucleocapsids in cells' cytoplasm and nucleus. PMID- 11878892 TI - Sequences and replication of genomes of the archaeal rudiviruses SIRV1 and SIRV2: relationships to the archaeal lipothrixvirus SIFV and some eukaryal viruses. AB - The double-stranded DNA genomes of the viruses SIRV1 and SIRV2, which infect the extremely thermophilic archaeon Sulfolobus and belong to the family Rudiviridae, were sequenced. They are linear, covalently closed at the ends, and 32,312 and 35,502 bp long, respectively, with an A+T content of 75%. The genomes of SIRV1 and SIRV2 carry inverted terminal repeats of 2029 and 1628 bp, respectively, which contain multiple direct repeats. SIRV1 and SIRV2 genomes contain 45 and 54 ORFs, respectively, of which 44 are homologous to one another. Their predicted functions include a DNA polymerase, a Holliday junction resolvase, and a dUTPase. The genomes consist of blocks with well-conserved sequences separated by nonconserved sequences. Recombination, gene duplication, horizontal gene transfer, and substitution of viral genes by homologous host genes have contributed to their evolution. The finding of head-to-head and tail-to-tail linked replicative intermediates suggests that the linear genomes replicate by the same mechanism as the similarly organized linear genomes of the eukaryal poxviruses, African swine fever virus and Chlorella viruses. SIRV1 and SIRV2 both contain motifs that resemble the binding sites for Holliday junction resolvases of eukaryal viruses and may use common mechanisms for resolution of replicative intermediates. The results suggest a common origin of the replication machineries of the archaeal rudiviruses and the above-mentioned eukaryal viruses. About 1/3 of the ORFs of each rudivirus have homologs in the Sulfolobus virus SIFV of the family Lipothrixviridae, indicating that the two viral families form a superfamily. The finding of inverted repeats of at least 0.8 kb at the termini of the linear genome of SIFV supports this inference. PMID- 11878893 TI - Induction of intestinal rotavirus-specific antibodies in respiratory, but not gut, lymphoid tissues following mucosal immunization of mice with inactivated rotavirus. AB - Intranasal (i.n.), but not oral, immunization of mice with inactivated rotavirus induces protection against challenge. To understand the mechanisms by which i.n. immunization with inactivated rotavirus evokes protective immunity, we examined the site of rotavirus-specific B cell activation and the origins of intestinal IgA-secreting B cells following i.n. inoculation of mice with inactivated rhesus rotavirus. We found that (1) i.n., but not oral, inoculation induced partial protection after challenge; (2) i.n., but not oral, inoculation induced production of rotavirus-specific IgM, IgA, and IgG by intestinal lymphoid tissues; and (3) after i.n. inoculation, nasal-associated lymphoid tissues (NALT) and bronchial lymph nodes (BLN) were the sites of initial production of rotavirus specific antibodies. These studies indicate that after inoculation with inactivated rotavirus, virus-specific effector B cells may be more easily activated in respiratory, compared to intestinal, lymphoid tissues. Additional studies are needed to determine if these observations are due to fundamental differences in the microenvironment of NALT and BLN compared to Peyer's patches or are a function of the anatomic differences between the respiratory and the gastrointestinal tracts. PMID- 11878894 TI - The Kaposi's sarcoma-associated herpesvirus latency-associated nuclear antigen binds to specific sequences at the left end of the viral genome through its carboxy-terminus. AB - Latent infection by members of the gammaherpesvirus family is typically characterized by stable episomal maintenance of genomic viral DNA. In the case of Epstein--Barr virus (EBV), this is dependent upon binding of the Epstein-Barr nuclear antigen 1 (EBNA1) to sites which lie within the origin of plasmid replication (OriP). The recently discovered Kaposi's sarcoma-associated herpesvirus (KSHV) encodes the latency-associated nuclear antigen (LANA), which appears to be important for supporting the latent infection of human cells by KSHV. The present work describes site-specific binding of the LANA protein to multiple different elements at the left end of the genome, a region which appears to be critical for maintenance of KSHV episomes. Of the three sites, terminal LANA-binding region 4 (TLBR4) binds LANA with the highest affinity when compared to the other sites. Further characterization of this cis-acting element by mutagenesis studies indicates that the minimal TLBR4-binding sequence is represented by a 13-bp sequence 5prime prime or minute CGCCCGGGCATGG 3prime prime or minute. Furthermore, this specific binding to TLBR4 was mediated by the distal 200 amino acid C-terminus of the LANA protein. PMID- 11878895 TI - Poliovirus 3C protease-mediated degradation of transcriptional activator p53 requires a cellular activity. AB - Infection of HeLa cells with poliovirus leads to rapid shut-off of host cell transcription by RNA polymerase II. Previous results have suggested that both the basal transcription factor TBP (TATA-binding protein) and transcription activator proteins such as CREB (cyclic AMP-responsive element-binding protein) and Oct-1 (the octamer-binding factor) are cleaved by the viral-encoded protease, 3C(Pro). Here we demonstrate that the transcriptional activator (and tumor suppressor) p53 is degraded by the viral protease 3C both in vivo and in vitro. Unlike other transcription factors that are directly cleaved by 3C(pro), degradation of p53 requires a HeLa cell activity in addition to 3C(Pro). The degradation of p53 by 3C(Pro) does not appear to involve the ubiquitin pathway of protein degradation. Vaccinia virus infection of HeLa cells leads to inactivation of the cellular activity required for 3C(Pro)-mediated degradation of p53. The vaccinia-encoded protein (CrmA) is known to inhibit caspase I (ICE protease) that converts inactive IL-1beta to an active secreted form. Incubation of HeLa cells with caspase I inhibitor Z-VAD-fmk does not interfere with 3C(Pro)-mediated degradation of p53. The cellular activity present in extracts of HeLa cells can be fractionated through phosphocellulose. A partially purified fraction that elutes at 0.6 M KCl from phosphocellulose contains the activity that degrades p53 in a 3C(Pro)-dependent manner. These results suggest that both poliovirus-encoded protease 3C(Pro) and a cellular activity are required for the degradation of p53 observed in cells infected with poliovirus. PMID- 11878896 TI - Expression of vaccinia E3L and K3L genes by a novel recombinant canarypox HIV vaccine vector enhances HIV-1 pseudovirion production and inhibits apoptosis in human cells. AB - Poxviruses that are attenuated for growth in human cells provide a safe means of HIV antigen expression and are capable of eliciting HIV-specific immune responses, including CD8+ cytotoxic T-lymphocyte (CTL) responses. HIV-1 antigen expression in human cells by attenuated poxvirus vectors may be limited by interferon-mediated host defense mechanisms. To enhance HIV antigen expression in human cells, the vaccinia virus E3L and K3L genes were inserted into a canarypox vector that expresses HIV-1 Gag, Env, and a Nef/Pol polyepitope string. E3L and K3L markedly reduced the activation of the double-stranded RNA-dependent protein kinase, PKR, and led to a significant reduction in apoptosis in HeLa cells. Production and release of HIV-1 antigen in the form of pseudovirions was enhanced in both duration and magnitude by this vector modification. The addition of immunomodulatory genes to attenuated poxviruses represents a novel strategy for enhancing antigen production by live vector HIV vaccine candidates. PMID- 11878897 TI - The small 11-kDa protein from B19 parvovirus binds growth factor receptor-binding protein 2 in vitro in a Src homology 3 domain/ligand-dependent manner. AB - The small 11-kDa proteins of B19 parvovirus contain three proline-rich regions which conform to consensus Src homology 3 (SH3) ligand sequences present in signaling molecules within the cell. We have shown that the B19 11-kDa proteins specifically interact with the growth factor receptor-binding protein 2 (Grb2) in vitro. Mutation of prolines within one of the three SH3 ligand-like sequences decreases the binding of B19 11-kDa proteins to Grb2, suggesting that the proline rich region is involved in the B19 11-kDa/Grb2 interaction. Therefore, the B19 11 kDa proteins may function to alter Grb2-mediated signaling by disrupting SH3 domain/ligand interactions. These results implicate the 11-kDa proteins in B19 pathogenesis through perturbation of normal cellular signaling pathways. PMID- 11878898 TI - p53-independent induction of apoptosis by the HTLV-I tax protein following UV irradiation. AB - Human T cell leukemia virus type 1 (HTLV-1) encodes a transforming protein, Tax. Tax is a promiscuous viral transactivator involved in both cell growth and death control. We have previously shown that Tax sensitizes cells to apoptosis induced by DNA-damaging agents and this report further characterizes the Tax-mediated apoptosis pathway. We found that Tax-mediated apoptosis in response to UV irradiation was inhibited by Bcl-2 and Bcl-X(L) overexpression and by treatment with the caspase inhibitor z-VAd-FMK. Since Tax has been shown to functionally inactivate the apoptosis regulator p53, the effect of Tax on apoptosis in the absence of p53 was examined. In these studies, Tax sensitized p53-negative cells to apoptose, suggesting that Tax can mediate a p53-independent form of apoptosis. In addition, cells expressing both Tax and p53 displayed higher levels of apoptosis than cells expressing either protein alone, suggesting that the apoptosis-inducing activities of Tax and p53 are not completely overlapping. These observations demonstrate that Tax can utilize a p53-independent mechanism to induce apoptotic cell death following UV irradiation. PMID- 11878899 TI - Membrane activity of the southern cowpea mosaic virus coat protein: the role of basic amino acids, helix-forming potential, and lipid composition. AB - Southern cowpea mosaic virus (SCPMV) is a spherical RNA virus with T = 3 icosahedral symmetry. The particle is composed of 180 subunits of the coat protein (CP) and one copy of the positive-sense viral RNA. The CP has two domains, the random (R) domain formed by the N-terminal 64 aa and the shell (S) domain (aa 65--260). The R domain is highly charged, with 11 of the N-terminal 30 residues being basic. It is localized to the interior of the native particle where it may interact with the viral RNA, but under certain pH and salt conditions the topology of the particle changes to externalize the R domain. Since the CPs of several spherical RNA viruses have been shown to interact with host membranes during infection, we have begun investigating the membrane interactions of the SCPMV CP using the artificial liposome membranes. Both the native CP and the R domain overexpressed in Escherichia coli were observed to interact with liposomes. The interaction between the R domain and liposomes required either anionic phospholipids or non-bilayer-forming lipids and involved electrostatic interactions since it was shown to be both pH and ionic strength dependent. The analysis of four different deletion and six different site directed substitution mutations partially mapped the region responsible for this interaction to residues 1--30. Analysis of this region of the R domain by circular dichroism indicated that it assumes an alpha-helical structure when exposed to liposomes composed of anionic lipids. Mutations, which extend the helical nature of this region, promoted an increased interaction. The possible role of the CP/lipid interaction in the SCPMV infection is discussed. PMID- 11878900 TI - Genetic analysis of bipartite geminivirus tissue tropism. AB - The bipartite geminiviruses bean golden mosaic virus (BGMV), cabbage leaf curl virus (CabLCV), and tomato golden mosaic virus (TGMV) exhibit differential tissue tropism in Nicotiana benthamiana. In systemically infected leaves, BGMV remains largely confined to vascular-associated cells (phloem-limited), whereas CabLCV and TGMV can escape into the surrounding mesophyll. Previous work established that TGMV BRi, the noncoding region upstream from the BR1 open reading frame (ORF), is required for mesophyll invasion, but the virus must also contain the TGMV AL23 or BL1/BR1 ORFs. Here we show that, in a BGMV-based hybrid virus, CabLCV AL23 also directed efficient mesophyll invasion in conjunction with TGMV BRi, which suggests that host-adaptation of AL23 is important for the phenotype. Cis-acting elements required for mesophyll invasion were delineated by analyzing BGMV-based hybrid viruses in which various parts of BRi were exchanged with those of TGMV. Interestingly, mesophyll invasion efficiency of hybrid viruses was not correlated with the extent of viral DNA accumulation. In conjunction with TGMV AL23, a 52-bp region of TGMV BRi with sequence homology to DNA A was sufficient for mesophyll invasion. This 52-bp sequence also directed mesophyll invasion in combination with the TGMV BL1/BR1 ORFs. Overall, these results are consistent with a model for mesophyll invasion in which AL2 protein, in association with host factors, acts through the 52-bp region in TGMV BRi to affect expression of the BR1 gene. PMID- 11878901 TI - Hybrid papillomavirus L1 molecules assemble into virus-like particles that reconstitute conformational epitopes and induce neutralizing antibodies to distinct HPV types. AB - Human papillomavirus (HPV) hybrid virus-like particles (VLPs) were prepared using complementary regions of the major capsid L1 proteins of HPV-11 and -16. These hybrid L1 proteins were tested for assembly into VLPs, for presentation and mapping of conformational neutralizing epitopes, and as immunogens in rabbits and mice. Two small noncontiguous hypervariable regions of HPV-16 L1, when replaced into the HPV-11 L1 backbone, produced an assembly-positive hybrid L1 which was recognized by the type-specific, conformationally dependent HPV-16 neutralizing monoclonal antibody (N-MAb) H16.V5. Several new N-MAbs that were generated following immunization of mice with wild-type HPV-16 L1 VLPs also recognized this reconstructed VLP, demonstrating that these two hypervariable regions collectively constituted an immunodominant epitope. When a set of hybrid VLPs was tested as immunogens in rabbits, antibodies to both HPV-11 and -16 wild-type L1 VLPs were obtained. One of the hybrid VLPs containing hypervariable FG and HI loops of HPV-16 L1 replaced into an HPV-11 L1 background provoked neutralizing activity against both HPV-11 and HPV-16. In addition, conformationally dependent and type-specific MAbs to both HPV-11 and HPV-16 L1 VLP were obtained from mice immunized with hybrid L1 VLPs. These data indicated that hybrid L1 proteins can be constructed that retain VLP-assembly properties, retain type-specific conformational neutralizing epitopes, can map noncontiguous regions of L1 which constitute type-specific conformational neutralizing epitopes recognized by N MAbs, and trigger polyclonal antibodies which can neutralize antigenically unrelated HPV types. PMID- 11878902 TI - Circular viral DNA and anomalous junction sequence in PBMC of HIV-infected individuals with no detectable plasma HIV RNA. AB - Closed circular (cc) forms of extrachromosomal HIV DNA are detected in patients with high viral loads; however, it is unclear whether these forms remain if virus replication is suppressed to undetectable levels by combination antiretroviral therapy. A nested primer polymerase chain reaction amplification assay was used to detect the presence of ccDNA containing two long terminal repeat sequences (2 LTR) in PBMC of patients with low or undetectable plasma HIV RNA. Fifty percent of patients with plasma RNA levels <50 copies/ml of blood had detectable 2-LTR DNA. Sequencing of the products identified normal LTR--LTR junctions in the minority of cases with the majority containing anomalies including deletions and insertions. The persistence of HIV ccDNA in patients with no detectable plasma RNA could be consistent with ongoing de novo infection of dividing cells or with stability of this form of DNA in nondividing cells. PMID- 11878903 TI - The effect of X4 and R5 HIV-1 on C, C-C, and C-X-C chemokines during the early stages of infection in human PBMCs. AB - To better define a mechanism underlying the increase in expression of certain proinflammatory chemokines during HIV-1 infection, we analyzed the effect of X4 HIV-1 infection on C, C-C, and C-X-C chemokine mRNA levels. We demonstrate that X4 HIV-1 infection augments the expression of RANTES, IP-10, MCP-1, and Ltn in peripheral blood mononuclear cells (PBMCs). R5 HIV-1 also induces an increase in both IP-10 and MCP-1 production. Binding of UV-inactivated HIV-1 elevates MCP-1, RANTES, MIP-1alpha, MIP-1beta, and IL-8 expression, but fails to alter the production of IP-10, suggesting that the induction of IP-10 is dependent on downstream events following viral internalization. Indeed, recombinant gp120 alone was able to stimulate an eightfold increase in MCP-1 expression, but was unable to induce any detectable increase in IP-10 protein. HIV-induced modulation of chemokine expression suggests a mechanism by which HIV-infected monocytes and T cells might recruit target cells to sites of active viral replication, thus potentially aiding in the spread of the virus. PMID- 11878904 TI - H5N1 influenza viruses isolated from geese in Southeastern China: evidence for genetic reassortment and interspecies transmission to ducks. AB - The H5N1 viruses (H5N1/97) associated with the "bird-flu" incident in the Hong Kong SAR have not been isolated since the slaughter of poultry in December 1997 brought that outbreak to an end. Recent evidence points to this virus as having arisen through a reassortment of a number of precursor avian viruses and a virus related to Goose/Guangdong/1/96 (H5N1) (Gs/Gd/96) was the likely donor of the H5 hemagglutinin. We characterize the Goose/Guangdong/1/96-like viruses isolated from geese and ducks imported into Hong Kong in the year 2000. Antigenically and genetically, these recent H5N1 viruses fall into two groups, one mainly associated with geese, and the other, recently transmitted to ducks. Further, viruses isolated from a goose and a duck in December 2000 have acquired NS, PA, M, and PB2 genes from the aquatic avian influenza gene pool through reassortment. For pandemic preparedness, it is important to monitor whether these reassortant viruses have the capacity for interspecies transmission to terrestrial poultry or mammals. PMID- 11878905 TI - Live and killed rhabdovirus-based vectors as potential hepatitis C vaccines. AB - A highly attenuated, recombinant rabies virus (RV) vaccine strain-based vector was utilized as a new immunization strategy to induce humoral and cellular responses against hepatitis C (HCV) glycoprotein E2. We showed previously that RV based vectors are able to induce strong immune responses against human immunodeficiency virus type I (HIV-1) antigens. Here we constructed and characterized three replication-competent RV-based vectors expressing either both HCV envelope proteins E1 and E2 or a modified version of E2 which lacks 85 amino acids of its carboxy terminus and contains the human CD4 transmembrane domain and the CD4 or RV glycoprotein cytoplasmic domain. All three constructs stably expressed the respective protein(s) as indicated by Western blotting and immunostaining. Moreover, surface expression of HCV E2 resulted in efficient incorporation of the HCV envelope protein regardless of the presence of the RV G cytoplasmic domain, which was described previously as a requirement for incorporation of foreign glycoproteins into RV particles. Killed and purified RV virions containing HCV E2 were highly immunogenic in mice and also proved useful as a diagnostic tool, as indicated by a specific reaction with sera from HCV infected patients. In addition, RV vaccine vehicles were able to induce cellular responses against HCV E2. These results further suggest that recombinant RVs are potentially useful vaccine vectors against important human viral diseases. PMID- 11878906 TI - Out-of-frame versus in-frame core internal deletion variants of human and woodchuck hepatitis B viruses. AB - Human hepatitis B virus (HBV) variants containing in-frame core internal deletion (CID) have been demonstrated to contain all the functional features of defective interfering (DI) particles (Yuan, T. T.-T., M.-H. Lin, D. S. Chen, and C. Shih, 1998, J. Virol. 72, 578-584). Here, we report that out-of-frame HBV CID variants exhibit defective interfering property similar to in-frame CID variants characterized previously. This result raises the possibility that it may be the deleted pregenomic RNA product, rather than the deleted core protein product, that is responsible for interference. Furthermore, a genomic deletion elsewhere does not cause interference since preS2 deletion variants exhibit no influence on wild-type HBV replication. Consistent with the natural occurrence of HBV CID variants, we recently identified CID variants of woodchuck hepatitis virus (WHV) in natural infection. However, unlike HBV CID variants, functional characterization of WHV CID variants using a human hepatoma cell line has not revealed any interference in tissue culture. In summary, defective interference is a general phenomenon for both in-frame and out-of-frame HBV CID variants. PMID- 11878907 TI - Hypermodification and immune escape of an internally deleted middle-envelope (M) protein of frequent and predominant hepatitis B virus variants. AB - Naturally occurring deletions within the human hepatitis B virus (HBV) preS2 region have frequently been identified in patients with hepatocellular carcinoma (HCC), while chronic carriers without cirrhosis and HCC contain no detectable preS2 deletion variants. We have characterized two different preS2 internal deletion variants from two patients. In addition to several weak phenotypes, our study revealed three unexpected strong phenotypes: (1) a paradoxical "hypermodification" phenomenon was observed with significantly increased size heterogeneity and molecular weights of the secreted middle (M) envelope proteins containing a preS2 internal deletion. This phenomenon was observed in transient transfection with a human hepatoma Huh7 cell line as well as in stable transfection with a rodent hepatoma cell line 7777. (2) A significantly increased intracellular accumulation of all three envelope proteins (large, middle, and small) was detected by both Western blot analysis and immunofluorescence microscopy. (3) The middle envelope proteins with a preS2 internal deletion were not recognized in vitro by a putative neutralizing antiserum, suggesting that these variants can evade immune recognition in vivo. To our knowledge, this is the first identification and characterization of the M deletion variant protein in HBV natural infection. PMID- 11878908 TI - The region of the HSV-1 latency-associated transcript required for epinephrine induced reactivation in the rabbit does not include the 2.0-kb intron. AB - Previous studies have localized the region of the latency-associated transcript (LAT) of HSV-1 responsible for epinephrine-induced reactivation in the rabbit eye model to the first 1.5 kb of the primary transcript. This region extends from the 5prime prime or minute exon of the primary LAT transcript through the 5prime prime or minute half of the LAT 2.0-kb intron. To determine whether the 5prime prime or minute end of the LAT intron contributes to the induced reactivation phenotype, three recombinant viruses containing deletions within this portion of the LAT intron were constructed. The three recombinants, containing deletions spanning a combined region of 969 bp at the 5prime prime or minute end of the LAT intron, reactivated with the wild-type frequency of 17syn+. These results indicate that the elements governing induced reactivation reside within the first 699 bp of the primary LAT transcript encoding the 5prime prime or minute LAT exon. PMID- 11878909 TI - Targeted therapy of respiratory syncytial virus in African green monkeys by intranasally administered 2-5A antisense. AB - Respiratory syncytial virus (RSV) is a leading cause of respiratory disease in infants, young children, immunocompromised patients, and the institutionalized elderly. Previous work had shown that RNase L, an antiviral enzyme of the interferon system, could be recruited to cleave RSV genomic RNA by attaching tetrameric 2prime prime or minute-5prime prime or minute-linked oligoadenylates (2-5A) to an oligonucleotide complementary to repetitive gene-start sequences within the RSV genome (2-5A antisense). A 2prime prime or minute-O-methyl RNA modified analog of the lead 2-5A anti-RSV chimera is shown here to have enhanced antiviral activity in cell culture studies while also cleaving RSV genomic RNA in an RNase L- and sequence-specific manner. When administered intranasally to RSV infected African green monkeys, this chimera reduced nasal RSV replication by up to four log(10) units in a dose- and time-dependent manner. PMID- 11878910 TI - Restriction of a Sindbis virus mutant in BHK cells and relief of the restriction by the addition of adenosine. AB - SV(PZF) is a mutant of Sindbis virus (SV) which we selected on the basis of its ability to replicate in mosquito cells treated with pyrazofurin (PZF), a drug which inhibits pyrimidine nucleotide biosynthesis (Lin et al., 2000, Virology 272, 61-71). Three mutations, A6627U, A7543U, and C7593A, were identified in the nsP4 (the viral RNA polymerase) coding region, which were required for the PZF resistant phenotype. We report here that SV(PZF) has a second phenotype. Its replication in BHK cells is severely restricted; yields of SV(PZF) from BHK cells are 100- to 1000-fold lower than the yields of standard SV (SV(STD)). However, addition of adenosine to the SV(PZF)-infected cultures completely relieves this restriction and results in yields comparable to those observed with SV(STD). Adenosine has no effect on the yield of SV(STD) from BHK cells. Synthesis of the viral structural proteins is markedly depressed in SV(PZF)-infected BHK cells, as is synthesis of the viral subgenomic (SG) RNA from which these proteins are translated. In contrast, normal amounts of genomic RNA are made. Experiments with mutagenized viruses indicated that the SV(PZF) mutation, C7593A, by itself, was sufficient to produce the restriction phenotype. However, this mutation not only changes Pro 609 of nsP4 to Thr, it also changes the nucleotide at the minus sign5 position of the SG promoter. To evaluate the relative contributions of the change in nsP4 and the change in the SG promoter to the restriction phenotype, we made use of double SG viruses, in which nsP4 and the promoter for the SG RNA which encodes the structural proteins can be changed independent of each other. Our results indicated that both the change in nsP4 and the change in the SG promoter were required to produce the full restriction phenotype. We suggest that the changes in nsP4 and the SG promoter destabilize the RNA initiation complex assembled at the SG promoter and that since ATP is the initiating nucleotide in the SG RNA transcript, the increased level of ATP resulting from the addition of adenosine is able to compensate for this destabilization and restore the synthesis of SG RNA to normal levels. PMID- 11878911 TI - HNF-3beta is a critical factor for the expression of the Jaagsiekte sheep retrovirus long terminal repeat in type II pneumocytes but not in Clara cells. AB - Jaagsiekte sheep retrovirus (JSRV) is the causative agent of ovine pulmonary adenocarcinoma (OPA), a sheep lung cancer that resembles human lung adenocarcinoma or bronchioloaveolar carcinoma (BAC). JSRV is the only retrovirus that shows lung tropism and induces pulmonary carcinoma. Several lines of evidence suggest that the lung tropism for JSRV is mainly determined by the viral long terminal repeats (LTR). In a previous study, we showed that HNF-3alpha and 3beta were able to transactivate the JSRV LTR when cotransfected into 3T3 cells. The JSRV LTR contains two putative HNF-3 binding sites; to investigate the contribution of each HNF-3 binding site to transcription, we generated reporter constructs with deletions or nucleotide substitutions in one or both of the putative HNF-3 binding sites. In murine MLE-15 cells (derived from type II pneumocytes), mutations within the upstream site (minus sign147 to minus sign128 bp) resulted in a 72% reduction of the LTR activity, while mutation of the downstream site had little effect. In contrast, transactivation of the JSRV LTR was greatly reduced in 3T3 cells cotransfected with an HNF-3alpha or -3beta expression plasmid when the downstream site was eliminated. Electrophoretic mobility shift assays (EMSA) revealed that nuclear extracts from MLE-15 cells, but not 3T3 cells, were able to form a retarded complex with oligonucleotides encompassing either the upstream or the downstream sites. Anti-HNF-3beta antiserum, but not anti-HNF-3alpha antiserum, supershifted both protein-DNA complexes. These results indicate that the JSRV LTR is activated by the lung specific transcription factor HNF-3beta and that the upstream HNF-3 binding site is essential for expression in MLE-15 cells. In contrast, transactivation by HNF 3beta in 3T3 cells is mediated through the downstream HNF-3 site. On the other hand, JSRV LTR expression in a mouse lung Clara cell-derived line (mtCC1-2) did not appear to be strongly dependent on either HNF-3 binding site. These results support the notion that JSRV lung tropism is determined by the transcriptional specificity of the JSRV LTR, which is governed by interactions with lung-specific transcription factors. PMID- 11878912 TI - The role of the CD4 receptor versus HIV coreceptors in envelope-mediated apoptosis in peripheral blood mononuclear cells. AB - We examined the role of CD4, CXCR4, and CCR5 in HIV envelope-mediated apoptosis by measuring the response of activated PBMCs to recombinant envelope proteins derived from CXCR4- and CCR5-utilizing viruses. Apoptosis of T cells was assessed by annexin-V staining and TdT-mediated dUTP-biotin nick-end labeling. Treatment of CCR5Delta32 homozygote PBMCs with a CCR5-specific envelope induced apoptosis in T cells, demonstrating that envelope--CD4 interactions are sufficient to induce apoptosis. However, a CXCR4-specific envelope induced higher levels of apoptosis than a CCR5-specific envelope, suggesting that envelope-mediated apoptosis can be enhanced by envelope--CXCR4 interactions. We conclude that envelope can induce apoptosis in T cells independently of the coreceptor specificity of a given envelope, or the expression profile of CXCR4 or CCR5 on a target cell. However, envelope--coreceptor interactions, and in particular, envelope--CXCR4 interactions, can contribute to this process. PMID- 11878913 TI - Insertion of HIV-1 genes into Ad4DeltaE3 vector abrogates increased pathogenesis in cotton rats due to E3 deletion. AB - Adenovirus type 5 (Ad5) E3 region proteins abrogate Ad pathogenicity in the lungs of cotton rats. Our use of Ad4-HIV E3-deleted (DeltaE3) recombinants as vaccines necessitates further examination of these viruses for enhanced pathogenesis. Equivalent infectious doses of Ad4 wild-type (Ad4WT), Ad4DeltaE3, and two recombinants: Ad4DeltaE3HIVenv and Ad4DeltaE3HIVgag, were inoculated intranasally into cotton rats. Ad4 viruses did not replicate in the lungs, but caused mild pathologic effects, including peribronchiolitis, bronchitis, alveolitis, and interstitial pneumonia. As found previously for Ad5, deletion of Ad4 E3 genes resulted in increased lung pathology. Surprisingly, insertion of HIV genes into this region significantly restored protection attributed to E3 gene products, diminishing overall pathologic effects to Ad4WT levels (P < 0.0001). Similarly, following administration of equivalent particle numbers of the four viruses, only Ad4DeltaE3 caused increased overall pathology, while the two HIV recombinant viruses showed effects comparable to Ad4WT (P < 0.01). Our observation that Ad4DeltaE3HIV recombinants are as safe in cotton rats as Ad4WT encourages their continued development as AIDS vaccines. PMID- 11878914 TI - The 3prime prime or minute-terminal structure required for replication of Barley yellow dwarf virus RNA contains an embedded 3prime prime or minute end. AB - We determined the 3prime prime or minute-terminal primary and secondary structures required for replication of Barley yellow dwarf virus (BYDV) RNA in oat protoplasts. Computer predictions, nuclease probing, phylogenetic comparisons, and replication assays of specific mutants and chimeras revealed that the 3prime prime or minute-terminal 109 nucleotides (nt) form a structure with three to four stem-loops followed by a coaxially stacked helix incorporating the last four nt [(A/U)CCC]. Sequences upstream of the 109-nt region also contributed to RNA accumulation. The base-pairing but not the sequences or bulges in the stems were essential for replication, but any changes to the 3prime prime or minute-terminal helix destroyed replication. The two 3prime prime or minute proximal tetraloops tolerated all changes, but the two 3prime prime or minute distal tetraloops gave most efficient replication if they fit the GNRA consensus. A mutant lacking the 3prime prime or minute-proximal stem-loop produced elevated levels of less-than-full-length minus strands, and no (+) strand. We propose that a "pocket" structure is the origin of (minus sign)-strand synthesis, which is negatively regulated by the inaccessible conformation of the 3prime prime or minute terminus, thus favoring a high (+)/(minus sign) ratio. This 3prime prime or minute structure and the polymerase homologies suggest that genus Luteovirus is more closely related to the Tombusviridae family than to other Luteoviridae genera. PMID- 11878915 TI - Four chimpanzee monoclonal antibodies isolated by phage display neutralize hepatitis a virus. AB - Chimpanzee immunoglobulins are virtually identical to human immunoglobulins and may have clinically useful applications. Four chimpanzee monoclonal antibodies (MAbs) to the hepatitis A virus (HAV) capsid were isolated from a combinatorial cDNA library of gamma1/kappa antibody genes using phage display. Competition assays indicated that three of the MAbs recognized the same or overlapping epitopes, whereas the fourth recognized a different, nonoverlapping epitope on the HAV capsid. All four MAbs neutralized the homologous HAV strain, HM-175, in a radioimmunofocus assay and two of the four MAbs neutralized a heterologous simian HAV strain, AGM-27. From these data, we conclude that the MAbs must recognize at least three epitopes on the HAV capsid. Furthermore, competition assays performed with neutralizing murine MAbs suggested that three of the chimpanzee MAbs recognized epitopes on the HAV capsid which have not been defined previously. PMID- 11878916 TI - Naturally occurring amino acid polymorphisms in human immunodeficiency virus type 1 (HIV-1) Gag p7(NC) and the C-cleavage site impact Gag-Pol processing by HIV-1 protease. AB - Human immunodeficiency virus type 1 (HIV-1) protease activity is targeted at nine cleavage sites comprising different amino acid sequences in the viral Gag-Pol polyprotein. Amino acid polymorphisms in protease and in regions of Gag, particularly p7(NC) and the C-cleavage site between p2 and p7(NC), occur in natural variants of HIV-1 within infected patients. Studies were designed to examine the role of natural polymorphisms in protease and to identify determinants in Gag that modulate protease processing activity. Closely related Gag-Pol regions from an HIV-1-infected mother and two children were evaluated for processing in an inducible expression system, for protease activity on cleavage site analogues, and for impact on replication by recombinant viruses. Gag-Pol regions displayed one of three processing phenotypes based on the appearance of Gag intermediates and accumulation of mature p24(CA). Gag-Pol regions that were processed rapidly to produce p24(CA) resulted in high-level replication by recombinant viruses, while slow-processing Gag-Pol variants resulted in recombinant viruses that replicated with reduced kinetics in both T cell lines and peripheral blood mononuclear cells. Direct impact by Gag sequences on processing by protease was assessed by construction of chimeric Gag-Pol regions and by site-directed mutagenesis. Optimal protease activity occurred when Gag and Pol regions were derived from the same gag-pol allele. Heterologous Gag regions generally diminished rates and extent of protease processing. Natural polymorphisms in novel positions in p7(NC) and the C-cleavage site have a dominant effect on protease processing activity. Accumulation of Gag products after processing at the C site appears to delay subsequent cleavage and production of mature p24(CA). PMID- 11878917 TI - Targeting retroviral vector infection to cells that express heregulin receptors using a TVA-heregulin bridge protein. AB - Previously we have shown that it is possible to target retroviral vectors to cells using avian sarcoma and leukosis virus (ASLV) receptor-ligand and receptor single-chain antibody bridge proteins (now designated as GATEs for guided adaptors for targeted entry). In this report we were interested in determining whether this approach can be used to deliver retroviral vectors specifically to cells that express heregulin receptors. Heregulin receptors are attractive targets for retroviral vector-based gene delivery protocols since they are often overexpressed on the surfaces of cancer cells. To explore this possibility, the TVA-herbeta1 protein was generated, consisting of the extracellular domain of the TVA receptor for ASLV-A fused to the EGF-like region of heregulin beta1. TVA herbeta1 bound specifically to cells that express heregulin receptors, rendering them susceptible to efficient and specific infection by subgroup A ASLV vectors. In addition, these activities of TVA-herbeta1 were abrogated specifically in the presence of another bridge protein that contained the same ligand domain. These data confirm that the GATE protein TVA-herbeta1 mediates targeted retroviral infection via cell surface heregulin receptors. PMID- 11878918 TI - Viral and cellular specificities of caprine arthritis encephalitis virus Vif protein. AB - The caprine arthritis encephalitis virus (CAEV) Vif protein is necessary for a productive infection of susceptible goat cells. The vif gene is conserved among all primate and most nonprimate lentiviruses. However, previous reports demonstrated that, in their respective host cells, primate Vif deleted lentiviruses could not be complemented by nonprimate Vif proteins, suggesting that species-specific restrictions between Vif and the virus-producing cells may modulate the Vif function on viral infectivity. Here we bring further support to this hypothesis since we show that CAEV Vif, when expressed in goat cells, is able to increase the infectivity of Vif deleted human immunodeficiency type-1 virus and of murine leukemia virus. Moreover, we demonstrate in vitro interactions between different Vif proteins and NC domains of heterologous Gag precursors, supporting the notion that species specificity of lentiviral infection is not due to molecular interactions between Vif and viral components. PMID- 11878919 TI - Heparan sulfate-mediated binding of infectious dengue virus type 2 and yellow fever virus. AB - Dengue virus type 2 and Yellow fever virus are arthropod-borne flaviviruses causing hemorrhagic fever in humans. Identification of virus receptors is important in understanding flavivirus pathogenesis. The aim of this work was to study the role of cellular heparan sulfate in the adsorption of infectious Yellow fever and Dengue type 2 viruses. Virus attachment was assessed by adsorbing virus to cells, washing unbound virus away, releasing cell-bound virus by freezing/thawing, and then titrating the released infectious virus. Treatment of cells by heparin-lyase, desulfation of cellular heparan sulfate, or treatment of the virus with heparin inhibited cell binding of both viruses. Heparin also inhibited Yellow fever virus infection by 97%. Using infectious virus, the present work shows the importance of heparan sulfate in binding and infection of these two flaviviruses. PMID- 11878920 TI - The most frequent naturally occurring length polymorphism in the HIV-1 LTR has little effect on proviral transcription and viral replication. AB - About 38% of primary HIV-1 long terminal repeats (LTRs) contain an insertion (consensus: 5prime prime or minute-ACYGCTGA-3prime prime or minute), termed the most frequent naturally occurring length polymorphism (MFNLP). The MFNLP binds several transcription factors and might affect HIV-1 replication and disease progression in infected individuals. However, its relevance for proviral transcription and for HIV-1 replication in primary cells is unclear. We utilized HIV-1 NL4-3 LTR variants to investigate the effect of the MFNLP on 5prime prime or minuteLTR transcriptional activity in various cell types. Notably, viral promoter activity was studied in primary cells in the context of the integrated provirus, using both single cycle assays with pseudotyped Luciferase reporter viruses and replication-competent HIV-1 mutants. Our results demonstrate that the presence, absence, or duplication of the 5prime prime or minute-ACYGCTGA-3prime prime or minute motif has little effect on viral promoter activity in T cell lines, peripheral blood mononuclear cells (PBMC), and monocyte-derived macrophages (MDM). Furthermore, all HIV-1 LTR variants showed efficient induction upon stimulation with TPA and/or ionomycin and replicated with comparable efficiency in a human T cell line and in PBMC. Thus, the MFNLP does not significantly affect HIV-1 5prime prime or minuteLTR transcriptional activity and viral replication in primary cells, suggesting that this common sequence variation has little impact on the clinical course of HIV-1 infection. PMID- 11878921 TI - Immunolocalization studies of an antisense protein in HIV-1-infected cells and viral particles. AB - The minus DNA strand of HIV-1 presents an open reading frame that is complementary to the HIV-1 envelope messenger, is highly conserved among HIV-1 isolates, and may encode a hydrophobic protein. In previous studies, the antisense transcript has been identified both in various HIV-infected cell lines and in leukocytes of HIV-1(+) patients. The expression of the corresponding antisense protein (ASP) during natural HIV-1 infection has been indirectly evidenced by the identification of anti-ASP antibodies in the sera of HIV(+) patients. We have used immunoelectron microscopy procedures (ultra-small gold particles coupled to silver enhancement) to establish direct evidence of ASP production. ASP has then been detected in chronically and acutely HIV-1-infected cell lines. The protein, found mostly associated with various cellular membranes as well as with the virions released from the cells, indicated that ASP is a bona fide component for the virions and may participate in the particle formation. PMID- 11878923 TI - Hepatitis C virus NS5A colocalizes with the core protein on lipid droplets and interacts with apolipoproteins. AB - The nonstructural protein 5A (NS5A) of the hepatitis C virus (HCV) has been shown to interact with a variety of cellular proteins and implicated in the regulation of cell growth, interferon resistance, and other cellular signaling pathways, but the role of NS5A in HCV pathogenesis has not been firmly established. To further characterize this multifunctional protein, we instigated the studies of the subcellular localization of NS5A in a hepatoma cell line. NS5A was localized to the perinuclear membrane structures, including the endoplasmic reticulum (ER) and the Golgi apparatus, by immunofluorescence staining and confocal microscopy. In addition, it was also associated with the surface of cytoplasmic globular structures when expressed alone or as a part of the NS3-5B polyprotein. Oil red O staining revealed that these globular structures were lipid droplets, where the HCV core protein was also localized. The association of NS5A with intracellular membrane was further confirmed by membrane flotation analysis. To determine whether NS5A interacts with any cellular lipid-binding protein, we performed yeast two-hybrid screening in both HepG2 and human liver cDNA libraries. Apolipoprotein A1 (apoA1), one of the protein components of high-density lipoprotein (HDL) particles, was identified by two independent screening processes. The interaction between NS5A and apoA1 was confirmed by both in vitro pull-down and in vivo coimmunoprecipitation experiments. Immunofluorescence staining revealed a significant colocalization of NS5A and apoA1 in the Golgi apparatus. Our results established an association of NS5A with lipid droplets and apoA1, suggesting that NS5A, together with the core protein, may play a role in the pathogenesis of the derangement of lipid metabolism, contributing to liver steatosis commonly observed in hepatitis C. PMID- 11878924 TI - A molecular switch in the capsid protein controls the particle polymorphism in an icosahedral virus. AB - The recombinant coat protein (CP) of Sesbania mosaic virus (SeMV; genus Sobemovirus) was found to self-assemble into capsids encapsidating 23S rRNA and CP mRNA in Escherichia coli. The CP lacking 22 amino acids from the N-terminus assembled into stable T = 3 capsids that appeared similar to SeMV, indicating that the N-terminal 22 amino acid residues are dispensable for T = 3 assembly. Two distinct capsids, T = 1 and pseudo T = 2, were observed when the N-terminal 36 amino acids encompassing the arginine-rich motif (N-ARM) were removed. Only T = 1 particles were observed upon deletion of 65 amino acids from the N-terminus, which also included the sequence element for the beta-annulus. These results reveal that N-ARM acts as a molecular switch in regulating T = 3 assembly. Formation of stable pseudo T = 2 particles shows that pentamers of AB dimers could nucleate assembly at icosahedral-5-folds. Capsids assembled from the N terminally truncated proteins also encapsidated 23S rRNA and CP mRNA, suggesting the presence of sites outside the N-terminal 65 residues that may be involved in RNA--protein interactions. PMID- 11878922 TI - Sequence analysis of the complete genome of an iridovirus isolated from the tiger frog. AB - We have isolated a tiger frog virus (TFV) from diseased tiger frogs, Rana tigrina rugulosa. The genome was a linear double-stranded DNA of 105,057 basepairs in length with a base composition of 55.01% G+C. About 105 open reading frames were identified with coding capacities for polypeptides ranging from 40 to 1294 amino acids. Computer-assisted analyses of the deduced amino acid sequences revealed that 39 of 105 putative gene products showed significant homology to functionally characterized proteins of other species in the GenBank/EMBL/DDBJ databases. These proteins included enzymes and structural proteins involved in virus replication, transcription, modification, and virus--host interaction. The deduced amino acid sequences of TFV gene products showed more than 90% identity to FV3, but a low degree of similarity among TFV, ISKNV, and LCDV-1. The results from this study indicated that TFV may belong to the genus Ranavirus of the family Iridoviridae. PMID- 11878925 TI - Caspase activation is required for permissive replication of Aleutian mink disease parvovirus in vitro. AB - Aleutian mink disease parvovirus (ADV) is distinct among the parvoviruses as infection in vivo is persistent, restricted, and noncytopathic. In contrast, infections with other more prototypic parvoviruses, like mink enteritis virus (MEV), are acute, cytopathic, and characterized by permissive replication in vivo. Although apoptosis results in the death of cells acutely infected by parvoviruses, the role of apoptosis in ADV infections is unknown. Permissive infection of ADV resulted in apoptosis of Crandell feline kidney (CrFK) cells as indicated by TUNEL staining, Annexin-V staining, and characteristic changes in cell morphology. Pretreatment of infected cells with caspase 3 or broad-spectrum caspase inhibitors prevented apoptosis. In addition, treatment of infected cells with these inhibitors caused a 2 log(10) reduction in the yield of infectious virus compared to untreated cultures. This block in replication preceded substantial viral DNA amplification and gene expression. However, inhibitors of caspases 1, 6, and 8 did not have this effect. MEV also induced caspase-dependent apoptosis following infection of CrFK cells, although production of infectious progeny was not affected by inhibition of apoptosis. Thus, permissive replication of ADV in vitro depended upon activation of specific caspases. If ADV infection of cells in vivo fails to initiate caspase activation, the requirement of caspase activity for replication may not be met, thus providing a possible mechanism for persistent, restricted infection. PMID- 11878926 TI - Marek's disease virus VP22: subcellular localization and characterization of carboxyl terminal deletion Mutations. AB - Marek's disease virus (MDV) is an alphaherpesvirus that causes T cell lymphoma and severe immunosuppression in chickens. The MDV UL49 gene, which encodes the tegument viral protein 22 (VP22), has been expressed as a green fluorescent protein (GFP) fusion protein in chicken embryonic fibroblasts to examine its subcellular localization. As with both human herpesvirus 1 and bovine herpesvirus 1VP22-GFP fusion proteins, the MDV VP22-GFP product binds to microtubules and heterochromatin. In addition, the MDV protein also binds to the centrosomes. During mitosis, VP22-GFP binds to sister chromatids, but dissociates from the centrosomes and the microtubules of the mitotic spindle. A series of VP22 carboxy terminal truncation mutants were constructed to define regions responsible for these binding properties. These mutants identified separable domains or motifs responsible for binding microtubules and heterochromatin. PMID- 11878927 TI - Site-directed mutagenesis of Avibirnavirus VP4 gene. AB - Virus protein VP4 of infectious bursal disease virus (IBDV) is a protease which separates VPX and VP3 from the polyprotein. We studied the importance of serine and aspartic acid on cleavage at the VPX/VP4 junction and analysed the role of the proposed H547, D590, and S653 catalytic site using five different mutations on VP4. Our results suggest that the replacement of serine by lysine in AXAAS motifs in serotype II IBDV influences polyprotein (PP) processing by VP4 and also indicate the presence of an alternative cleavage site. Mutation on D ((510)TLAADK(515)) prevented the cleavage at the VPX/VP4 junction, but we have found that independently of the importance of those alanines in LAA, D has an important role as part of the cleavage site. Replacement of histidine by proline H547P completely abolished PP processing. Mutation on D590 induced a partial PP processing when it was replaced by proline and the replacement of serine by proline at S653P induced a prominent change in PP processing. These results permit us to conclude that IBDV VP4 has the ability to act according to structural and topographical changes during translational and posttranslational processes and allow multiple hit sites, which serve to increase effectiveness. PMID- 11878928 TI - Genetic constraints and the adaptive evolution of rabies virus in nature. AB - We used a molecular evolutionary approach to investigate the species adaptation of rabies virus in nature. A maximum likelihood analysis of selection pressures revealed that the nucleoprotein (N) and glycoprotein (G) genes of natural viral isolates were highly constrained, especially at nonsynonymous sites, in contrast to the higher rates of nonsynonymous evolution observed in viruses subject to laboratory passage. Positive selection was only found at a single amino acid site -position 183 in the ectodomain of the G gene. The low rate of nonsynonymous evolution in natural isolates of rabies virus may be due to constraints imposed by the need to replicate in multiple cell types within the host, which in turn facilitates cross-species transmission, or because viral proteins are not subject to immune selection. Using known dates in the epidemiologic history of European viral isolates, we estimated that overall rates of nucleotide substitution in rabies virus were similar to those observed in other RNA viruses. Assuming that the average rate of synonymous change does not vary among species, we estimated that the current genetic diversity in lyssavirus genotype 1 may have arisen only during the last 500 years. PMID- 11878929 TI - Induction of Bad-mediated apoptosis by Sindbis virus infection: involvement of pro-survival members of the Bcl-2 family. AB - It is known that infection with Sindbis virus (SNV) induces apoptosis, which is inhibited by two pro-survival members of the Bcl-2 family, Bcl-2 and Bcl-xL. However, the mechanism of involvement of the other members of the Bcl-2 family in SNV-induced apoptosis remains unclear. In this study we report that Bad protein, one of the pro-apoptotic Bcl-2 family members, mediates apoptosis in the mammalian cells infected with SNV. Expression of Bad was shown to promote SNV induced apoptosis in human embryonic kidney 293T and baby hamster kidney cells. SNV infection also induced translocation of endogenous Bad into mitochondria and heterodimerization of Bad with Bcl-xL. On the other hand, the structurally most similar pro-survival members, Bcl-2, Bcl-xL, and Bcl-w, suppressed SNV-induced apoptosis in the absence of Bad, whereas Mcl-1 and A1 did not. Bcl-w could inhibit SNV-induced apoptosis in the presence of Bad, but Bcl-xL could not. Bad could be coimmunoprecipitated with Bcl-xL or Bcl-2, but not with Bcl-w. Two viral Bcl-2 homologs, E1B19K and BHRF1, also suppressed SNV-induced apoptosis irrespective of the presence of Bad and no physical association with Bad was observed. These results suggest that direct interaction of Bad with pro-survival members of the Bcl-2 family contributes to the progress of SNV-induced apoptosis and that nonbinding members restrain SNV-induced apoptosis irrespective of Bad expression. PMID- 11878930 TI - Hepatitis C virus core protein induces cell proliferation and activates ERK, JNK, and p38 MAP kinases together with the MAP kinase phosphatase MKP-1 in a HepG2 Tet Off cell line. AB - Hepatitis C virus (HCV) core protein is a multifunctional protein interacting with cellular and viral proteins and promoters. A tetracycline-regulated system was used to generate a HepG2 Tet-Off cell line allowing regulated expression of a full-length (191 aa) and an N(c)-truncated core protein (160 aa). In this system HCV core protein expression activates extracellular signal-regulated kinase (ERK), c-jun N-terminal kinase (JNK), and p38 mitogen-activated protein (MAP) kinase, induces MAP kinase phosphatase MKP-1 expression, and increases cell proliferation. This was accompanied by an activation of c-Jun and ATF-2, but not Elk-1 and c-Fos. Furthermore, AP-1 activation was independent of c-Fos. Full length and N(c)-truncated HCV core proteins exerted similar effects. PMID- 11878931 TI - Vaccinia virus CrmE encodes a soluble and cell surface tumor necrosis factor receptor that contributes to virus virulence. AB - Poxviruses encode soluble cytokine receptors to interfere with host immune functions. Cells infected with vaccinia virus (VV) strains USSR, Lister, and Evans express soluble and cell surface tumor necrosis factor receptors (vTNFRs). We have characterized vTNFR activity in VV USSR and identified an open reading frame that encodes both soluble and cell surface activity, hereafter referred to as VV cytokine response modifier E (VV CrmE). Expression and characterization from recombinant VV and baculovirus showed VV CrmE to be an 18-kDa protein that bound human, mouse, and rat TNF-alpha, but not human LT alpha. VV CrmE inhibited the cytotoxic and apoptotic activities of human, but not mouse or rat, TNF in vitro. Nonetheless, in a murine intranasal model, USSR recombinants lacking CrmE were attenuated, demonstrating a role in vivo. Furthermore, expression of VV or cowpox virus vTNFRs from VV strain WR (which itself does not express a vTNFR) was shown to enhance virulence in the murine model. PMID- 11878932 TI - Genome organization of Casphalia extranea densovirus, a new iteravirus. AB - The viral genome of Casphalia extranea densovirus (CeDNV) has been cloned and sequenced. It was 5002 nucleotides long and contained inverted terminal repeats of 230 nucleotides. Their distal 159 nucleotides formed imperfect palindromes in two orientations. Three large open reading frames (ORFs) were identified on the same strand, two in the left-hand half and one in the right-hand half. Each of the five structural proteins, expressed from the right-hand ORF in the baculovirus system, autoassembled into capsids. The two left-hand ORFs overlapped and code for nonstructural (NS) proteins. NS1 protein was shown to contain replicator protein and helicase/ATPase motifs. The PGY region in VP1 capsid protein is conserved among most parvoviruses and contained a phospholipase A(2) motif, a novel viral enzyme. This domain was expressed and its enzyme activity was demonstrated. The approximate 75% sequence identity between the DNAs from CeDNV and BmDNV-1 and identical genome organization indicated that CeDNV should be classified in the Iteravirus genus. PMID- 11878934 TI - Hypophosphorylation of poly(A) polymerase and increased polyadenylation activity are associated with human immunodeficiency virus type 1 Vpr expression. AB - The HIV-1 encoded accessory protein, viral protein R (Vpr) is responsible for several biological effects in HIV-1-infected cells including nuclear transport of the preintegration complex, activation of long terminal repeat (LTR)-mediated transcription, and the induction of cell-cycle arrest and apoptosis. Vpr's ability to arrest cells at the G2 phase of the cell cycle is due to the inactivation of p34(cdc2) cyclin B complex, resulting in hypophosphorylation of substrates involved in cell-cycle progression from G2 to mitosis (M). Poly(A) polymerase (PAP), the enzyme responsible for poly(A) addition to primary transcripts, contains multiple consensus phosphorylation sites for p34(cdc2) cyclin B kinase that regulates its catalytic activity. We investigated the effects of Vpr on the activity of PAP in Jurkat cells using a superinfection system. Superinfection of cells using Vpr+ vesicular stomatitis virus G protein (VSV-G)-pseudotyped virus caused a complete dephosphorylation of PAP. Cotransfection studies in 293T cells and Xenopus oocyte RNA injection experiments mirrored these effects. Vpr's dramatic effect on PAP dephosphorylation was reflected in enhanced polyadenylation activity in PAP activity assays. HIV-1 Vpr appears to enhance processes that are coupled to transcription such as polyadenylation and could ultimately prove to optimize HIV-1 replication and contribute to HIV-1 pathogenesis. (C)2002 Elsevier Science. PMID- 11878935 TI - Abrupt interfacial transitions of hydrophobic polysilanes as probed via liquid crystal-assisted stepwise deposition. AB - The Langmuir-Blodgett deposition was applied for the first time for fully hydrophobic polysilanes, poly(di-n-hexylsilane) (PDHS) and poly(methyl-n octylsilane) (PMOS). The monolayer formation of such hydrophobic polysilanes was achieved by cospreading with a liquid crystal molecule, 4'-pentyl-4-cyanobiphenyl (5CB). The UV-visible absorption spectra showed that the backbone conformations of the polysilanes are completely different from that in the bulk. The layer-by layer deposition allowed for the nanometer level evaluation of the thickness profile on the conformational changes. It is found that the conformational change of these hydrophobic polysilanes abruptly occurs at a monolayer level thickness boundary. PMID- 11878933 TI - Inhibition of serum- and calcium-induced terminal differentiation of human keratinocytes by HPV 16 E6: study of the association with p53 degradation, inhibition of p53 transactivation, and binding to E6BP. AB - Transfection of the E6 gene of human papillovirus (HPV) 16 into primary human keratinocytes (PHKs) generates proliferating cell colonies which are resistant to serum- and calcium-induced terminal differentiation. The extreme C-terminus of E6 was shown to be dispensable for this activity. To map further the amino acid sequences required for inducing resistance to serum and calcium, and to address the functional significance of E6 interactions with p53 and E6BP (ERC-55) in this function, we evaluated the activities of a series of E6 mutants. Small deletions within the central portion of the second putative zinc-finger abolished, or markedly reduced, E6 biological activity, while mutations affecting the cysteine residues in the base of the finger were less effective in this respect. When these mutants were assayed for their ability to degrade p53 in vitro and in vivo and to inhibit p53 transcriptional activation (TA), we found that there was a dissociation of these activities in some mutants. We mapped one mutant which was highly efficient in p53 degradation and inhibition of p53 TA, yet displayed severely reduced activity in the biological assay, and conversely, a subset of mutants that showed moderate activities in the colony assay while being severely impaired in p53 degradation and inhibition of p53 TA. These data argue that p53 inactivation or even elimination are not sufficient, and may not be essential, for altering the response of PHKs to serum and calcium. When these E6 mutants were evaluated for E6BP binding in vitro, there was a similar dissociation between the biological and biochemical activities of several mutants. We mapped mutants with moderate activity in the biological assay that lacked the ability to bind to E6BP and a mutant that showed high biological activity with only marginal binding to E6BP. Thus, there is no absolute correlation between the ability of E6 mutant proteins to induce alterations in keratinocyte differentiation responses to calcium and serum and to induce p53 degradation, inhibit p53 mediated transactivation, or bind E6BP. Evidently there are additional cellular targets for E6 which mediate this alteration in cellular differentiation. PMID- 11878936 TI - Highly selective excitation in biomolecular NMR by frequency-switched single transition cross-polarization. AB - A new method for selective excitation in biomolecular NMR uses two-fold single transition cross-polarization between protons and nitrogen-15 or carbon-13 nuclei. Switching the frequencies between the forward and backward transfer steps allows one to select a multiplet pattern that is associated with a single pair of spins in a medium-size protein. The efficiency of the transfer benefits from so called TROSY line-narrowing effects which arise from interference between relaxation mechanisms. PMID- 11878937 TI - A polar effects controlled enantioselective 1,2-chlorine atom migration via a chlorine-bridged radical intermediate. AB - An enantioselective 1,2-chlorine atom migration was observed in the tributyltin hydride reduction of various dihalogenated dihydrocinnamic acid derivatives. It is proposed that the reduction involves the formation of a chlorine-bridged radical intermediate, followed by hydrogen atom transfer to either the beta- or the alpha-carbon. The product distribution is affected by electron-withdrawing groups in that hydrogen atom transfer to the proximate carbon is favored. Presumably, this is due to the transition state of the hydrogen delivery step being electron rich, due to the relatively electropositive tin radical. These results demonstrate a 1,2-chlorine atom migration reaction governed by polar effects. PMID- 11878938 TI - The total synthesis of (+/-)-merrilactone A. AB - The total synthesis of the title compound has been accomplished in 20 steps. The key step is a free radical cyclization of vinyl bromide 29 to afford 30. The synthesis also features an efficient Dielsminus signAlder reaction of 2,3 dimethylmaleic anhydride with 1-(tert-butyldimethylsiloxy)-butadiene. The oxetane moiety of merrilactone A is fashioned via a Payne-like rearrangement of a hydroxyepoxide (see 2 right arrow 1). PMID- 11878939 TI - Salt effects on solute exchange in sodium dodecyl sulfate micelles. AB - We describe the influence of sodium chloride on the rate of solute exchange in aqueous SDS micelles for a water-insoluble solute, a pyrene-containing triglyceride 1. The initially prepared solutions contained a small fraction of micelles containing two molecules of 1 and a large excess of empty micelles. These solutions showed a measurable excimer emission (of intensity I(E)) that was stable for days to weeks in the absence of added salt. Following additions of salt, I(E) decayed exponentially (rate constant, k(obs)) accompanied by an increase in pyrene monomer emission. Values of k(obs) increased strongly with ionic strength (k(obs) similar [Na(+)](4)). There was no contribution of the empty micelle concentration beyond its contribution to the sodium ion concentration. We conclude that the solute exchange involves spontaneous fragmentation of the SDS micelles into two submicelles, each bearing a molecule of 1, which then grow back to normal micelles through condensation of SDS monomers. We propose a model for the fragmentation process in which large amplitude surface fluctuations "pinch off" a subunit that becomes a submicelle. These fluctuations bring sulfate headgroups into close proximity. Fluctuations leading to fission become important only in the presence of sufficient counterion concentration to reduce the electrostatic repulsion between neighboring headgroups. PMID- 11878940 TI - An engineered azurin variant containing a selenocysteine copper ligand. AB - Modulating the properties of proteins through de novo design or redesign of existing proteins has been a longstanding goal in protein chemistry. Over the past two decades, site-directed mutagenesis has been a powerful tool to probe the role of certain residues and to fine-tune the activity of proteins. A limitation of this approach has been the accessibility of only a restricted number of functional groups through the 20 amino acids in the genetic code. The more recent technique of expressed protein ligation (EPL) provides an alternative route that allows efficient incorporation of nonnatural residues into proteins. We report here the preparation and spectroscopic characterization of an azurin variant in which a cysteine ligand to the blue copper center has been replaced by EPL with selenocysteine (Sec). This reports marks the first time that selenocysteine is artificially incorporated into the active site of a metalloprotein. The variant displays a significantly increased A(parallel) (from 56 to 104 G) and red-shifted CT band (from 625 to 677 nm), while maintaining the general type 1 copper characteristics, including similarity in reduction potentials. This study illustrates that iso-structural substitution using EPL can fine-tune the structural and functional properties of a metal-binding site without loss of most of its characteristics. Further spectroscopic and X-ray crystallographic studies of this and other EPL variants will provide new insights into the fine-control of the structure and function of metalloproteins. PMID- 11878941 TI - Photoreactivity of 5-iodouracil-containing DNA-Sso7d complex in solution: the protein-induced DNA kink causes intrastrand hydrogen abstraction from the 5 methyl of thymine at the 5' side. AB - Photoirradiation of 5-iodouracil-containing DNA, d(GTAAT(I)UAC)(2) with Sso7d protein, possessing significant kink in DNA in the crystal structure induces an unprecedented intrastrand H abstraction at the methyl group of T(5), together with selective photooxidations at Met29 of Sso7d. The reactivity of the deoxyuridin-5-yl radical can be explained by the crystal structure of the d(GTAATTAC)(2)-Sso7d complex, suggesting that the interaction of DNA-Sso7d in solution is substantially similar to its crystal structure. PMID- 11878942 TI - A practical enantioselective fluorescent sensor for mandelic acid. AB - A novel optically active bisbinaphthyl fluorescent sensor, (S,S)- or (R,R)-1, is designed for the recognition of chiral alpha-hydroxycarboxylic acids. A convenient method has been developed to synthesize this compound. It is observed that (S)-mandelic acid enhances the fluorescence intensity of the (S,S)-sensor significantly more than (R)-mandelic acid does. The enantioselective fluorescent response is confirmed with the observation of a mirror image relationship for the interaction of (S,S)- and (R,R)-sensors with mandelic acid. The enantioselectivity in fluorescence response [(I(S) - I(0))/(I(R) - I(0)) = 2.49] is quite high, which makes the sensor useful for practical application. The fluorescence intensity change of the sensor is found to be linearly related to the enantiomeric composition of mandelic acid. This sensor is potentially useful for the combinatorial search of chiral catalysts for the asymmetric synthesis of alpha-hydroxycarboxylic acids. PMID- 11878943 TI - Site and oxidation-state specificity yielding dimensional control in perovskite ruthenates. AB - Sr(3)CaRu(2)O(9), a new 2:1 B-site ordered perovskite ruthenate, was synthesized and its structure determined based on powder X-ray, neutron and electron diffraction data. It is composed of one layer of CaO(6) alternating with two layers of RuO(6) perpendicular to the [111] axis of the cubic perovskite structure. The ordering leads to a [-Ru-Ru-Ca-] repeat unit along each of the pseudocubic directions. Sr(3)CaRu(2)O(9) is the first example of this structure type to include a majority metal with d electrons (Ru(V), d(3)). Three dimensional Sr(3)CaRu(2)O(9) can be transformed to the layered Ruddlesden-Popper phase Sr(1.5)Ca(0.5)RuO(4) (i.e., Sr(3)CaRu(2)O(8)) by reduction at 1200 degrees C in flowing argon. The original structure can be restored by oxidation of Sr(1.5)Ca(0.5)RuO(4) at 1000 degrees C in flowing oxygen. This remarkable transformation highlights the structural versatility afforded by the combination of ruthenium and calcium. PMID- 11878944 TI - Iridium-catalyzed H/D exchange into organic compounds in water. AB - The air-stable complex Cp(PMe(3))IrCl(2) efficiently catalyzes the exchange of deuterium from D(2)O into both activated and unactivated C-H bonds of organic molecules without added acid or stabilizers. Selectivity is observed in many cases, with activation of primary C-H bonds occurring preferentially. A number of new stoichiometric transformations involving the iridiym catalyst precursor are also presented, including an oxidation-decarbonylation reaction with primary alcohols. PMID- 11878945 TI - Unanticipated guest motion during a phase transition in a ferroelastic inclusion compound. AB - Urea inclusion compounds (UICs) have been used as tools to understand ferroelastic domain switching and molecular recognition during crystal growth. Although the vast majority of UICs contain helical arrangements of host H-bonds, those containing guests with the formula X(CH(2))(6)Y (X, Y = Br, Cl, CN, NC) adopt an alternative P2(1)/n packing mode in which the host molecules exist as stacked loops of urea hexamers. Such structures may be further separated into two classes, ones distorted away from hexagonal symmetry along [100] (Br(CH(2))(6)Br, Br(CH(2))(6)Cl, and Cl(CH(2))(6)Cl) and those distorted along [001] (e.g. NC(CH(2))(6)CN). In each of these systems, guests exist as equilibrium mixtures of gauche conformers whose populations control the direction and magnitude of the observed distortion. Such UICs are potentially ferroelastic, but the n-glide requires that domains are not related by a simple rotation-translation mechanism as in the helical systems. Ferroelastic (degenerate) domain reorientation would necessitate a large-scale reorganization of the urea framework and rupture of numerous H-bonds. Coupled with distortions of 2 to 10%, this mechanism-based barrier to domain switching has precluded observation of this phenomenon. To prepare ferroelastic UICs with minimal distortions from hexagonal symmetry, attempts were made to form solid solutions of UICs containing guests from the two classes. This failed, however: solid solution formation of the stacked loop form is usually possible within a series (e.g. with Cl(CH(2))(6)Cl and Br(CH(2))(6)Br), but not between series (e.g. Cl(CH(2))(6)Cl and NC(CH(2))(6)CN). Crystals of Cl(CH(2))(6)CN/urea, in which a single guest contains substituents from each class, are distorted along [001] by only 0.5% from hexagonal symmetry at 298 K and exhibit ferroelastic domain reorientation at high forces. At -66 degrees C, Cl(CH(2))(6)CN/urea undergoes a topotactic phase transition that is unexpectedly nontopochemical. The structure of the low-temperature phase, including the orientation of the methylene chain, closely matches the structures of UICs distorted by 10% along [100] (e.g. Cl(CH(2))(6)Cl/urea). In this transition, small conformation changes of guests give rise to large-scale guest translations of approximately 5.5 A down the channel axis, even though an analogous gauche-to-gauche jump is well established in closely related materials that adopt either high- or low-temperature forms (e.g. NC(CH(2))(6)CN/urea, Cl(CH(2))(6)Cl/urea). The large guest displacement during this transition explains the difficulty in preparing solid solutions of the P2(1)/n form with guests of formula X(CH(2))(6)Y from two different series (e.g. Cl(CH(2))(6)Cl and NC(CH(2))(6)CN). This failure arises not from the different orientations of guest induced strain, but from preferential occupation of different sites along the channel by the two types of guests. The subtlety of this process and of the interactions involved highlights the difficulty in using simple considerations of isomorphism to design new materials. PMID- 11878946 TI - Quencher as leaving group: efficient detection of DNA-joining reactions. AB - We describe a new fluorescence reporting strategy in which dabsyl, a well-known quencher, activates a hydroxyl group in a probe to convert it to a leaving group. When a nucleophilic phosphorothioate probe binds adjacent to a dabsyl quenched probe, autoligation occurs, releasing the quencher, and lighting up the probes, This signal change can be used to detect single nucleotide differences in DNA without enzymes or reagents. PMID- 11878947 TI - Telomestatin, a potent telomerase inhibitor that interacts quite specifically with the human telomeric intramolecular g-quadruplex. AB - Telomestatin is a natural product isolated from Streptomyces anulatus 3533-SV4 and has been shown to be a very potent telomerase inhibitor. The structural similarity between telomestatin and a G-tetrad suggested to us that the telomerase inhibition might be due to its ability either to facilitate the formation of or trap out preformed G-quadruplex structures, and thereby sequester single-stranded d[T(2)AG(3)](n) primer molecules required for telomerase activity. Significantly, telomestatin appears to be a more potent inhibitor of telomerase (5 nM) than any of the previously described G-quadruplex-interactive molecules. In this communication we provide the first experimental evidence that telomestatin selectively facilitates the formation of or stabilizes intramolecular G-quadruplexes, in particular, that produced from the human telomeric sequence d[T(2)AG(3)](4). A simulated annealing (SA) docking approach was used to study the binding interactions of telomestatin with the intramolecular antiparallel G-quadruplex structure. Each intramolecular G quadruplex molecule was found to bind two telomestatin molecules (unpublished results). A 2:1 model for the telomestatin bound in the external stacking mode in an energy minimized complex with the human telomeric basket-type G-quadruplex was constructed. Our observation that a G-quadruplex-interactive molecule without significant groove interactions is able to reorient in a G-quadruplex structure proints to the importance of core interaction with an asymmetric G-quadruplex structure in producing selective binding. Furthermore, the G-quadruplex interactions of telomestatin are more selective for the intramolecular structure in contrast to other G-quadruplex-interactive agents, such as TMPyP4. PMID- 11878948 TI - Micropatterning of polymer thin films with pH-sensitive and cross-linkable hydrogen-bonded polyelectrolyte multilayers. AB - Polyelectrolyte multilayers of poly(acrylic acid) (PAA) and polyacrylamide (PAAm) were prepared via hydrogen-bonding interactions. These multilayers as assembled were stable at low pH but dissolved quickly in neutral pH water. We developed methods for stabilizing these multilayers to high pH through cross-linking by heating or UV-irradiation. Thermal treatment of the multilayers, which resulted in a partial imidization between carboxylic acid and amide groups, gave the multilayer good stability at high pH. In addition, we introduced photoreactive groups in the multilayer, which rendered the film insoluble after UV irradiation. Using these selective stabilization approaches, we have succeeded in micropatterning these films by ink-jet printing and photolithography to create subtractive patterns. PMID- 11878949 TI - Intramolecular silicon-assisted cross-coupling reactions: general synthesis of medium-sized rings containing a 1,3-cis-cis diene unit. AB - The combination of ring-closing metathesis and Pd-catalyzed, silicon-assisted intramolecular cross-coupling has been developed to provide an effective and powerful method for construction of medium-sized rings with an internal 1,3-cis cis diene unit. Allylic alcohols bearing a Z-iodoalkenyl tether can be silylated with chlorodimethylvinylsilane and subjected to Mo-catalyzed ring-closing metathesis to form unsaturated siloxanes. Activation of the siloxane with tetrabutylammonium fluoride in the presence of [allylPdCl](2) leads to high yielding ring-closing reactions to form 9-, 10-, 11- and 12-membered rings. Extension to the synthesis of 9-membered ring unsaturated ethers has also been accomplished. Noteworthy features of this process include: (1) a highly stereospecific intramolecular coupling process, (2) flexible positioning of the hydroxy group, and (3) potential extension to other medium-sized carbocycles and heterocycles. PMID- 11878950 TI - Structures of protein-protein complexes are docked using only NMR restraints from residual dipolar coupling and chemical shift perturbations. AB - NMR structures of protein-protein and protein-ligand complexes rely heavily on intermolecular NOEs. Recent work has shown that if no significant conformational changes occur upon complex formation residual dipolar coupling can replace most of the NOE restraints in protein-protein complexes, while restraints derived from chemical shift perturbations can largely replace intermolecular NOEs in protein ligand structures. By combining restraints from chemical shift perturbations with orientation restraints derived from measurements of residual dipolar couplings, we show that the structure of the EIN-HPr complex can be calculated without NOE restraints. The final structure, built from the crystal structures of EIN and HPr in their uncomplexed form and docked only with NMR restraints, places HPr within 2.5 A of the position determined from the mean NMR structure of the complex. PMID- 11878951 TI - A new two-step four-component synthesis of highly functionalized cyclohexenols by sequential nickel-catalyzed couplings. AB - A new two-step procedure for the synthesis of cyclohexenols has been developed. A nickel-catalyzed three-component addition of an enal, alkyne, and acetylenic tin affords substituted hept-4-en-6-ynals. The products of this first step then undergo a second nickel-catalyzed reaction with organozincs or organoboranes to afford densely functionalized cyclohexenols. Variation in each of the four components is tolerated to provide access to a wide range of versatile building blocks. PMID- 11878952 TI - beta-diketiminate ligand backbone structural effects on Cu(I)/O(2) reactivity: unique copper-superoxo and bis(mu-oxo) complexes. AB - Copper(I) and -(II) complexes of beta-diketiminate ligands with identical flanking 2,6-diisopropylphenyl groups but divergent backbone substitution patterns were prepared and structurally characterized, and reactions of the Cu(I) species with O(2) at low temperature were explored. Despite being far removed from the coordinated metal ion, the different backbone patterns significantly influence the steric encumbrance exerted by the ligands, as revealed by differences in (a) the structural features of the Cu(I) and Cu(II) complexes and (b) the course of the oxygenation reactions of the Cu(I) compounds. With the less hindered ligand, a rare example of a neutral bis(mu-oxo)dicopper complex was identified on the basis of its diagnostic spectral features (UV-vis, resonance Raman, EPR) and the stoichiometry of O(2) uptake (Cu:O(2) = 2:1). In contrast, oxygenation of the Cu(I) complexes supported by the more hindered ligands yielded novel (superoxo)copper complexes, identified by a Cu:O(2) ratio of 1:1, a lack of an EPR signal, and O-isotope sensitive resonance Raman spectral features (nu(O)( )(O) = 968 cm(-1), Delta(18)O(2) = 51 cm(-1)). Symmetric coordination of the superoxo ligand is proposed on the basis of Raman data acquired using (16)O(18)O (single peak at 943 cm(-1)). PMID- 11878953 TI - Cyanide-limited complexation of molybdenum(III): synthesis of octahedral [Mo(CN)(6)](3-) and cyano-bridged [Mo(2)(CN)(11)](5-). AB - Octahedral coordination of molybdenum(III) is achieved by limiting the amount of cyanide available upon complex formation. Reaction of Mo(CF(3)SO(3))(3) with LiCN in DMF affords Li(3)[Mo(CN)(6)] x 6DMF (1), featuring the previously unknown octahedral complex [Mo(CN)(6)](3-). The complex exhibits a room-temperature moment of mu(eff) = 3.80 mu(B), and assignment of its absorption bands leads to the ligand field parameters Delta(o) = 24800 cm(-1) and B = 247 cm(-1). Further restricting the available cyanide in a reaction between Mo(CF(3)SO(3))(3) and (Et(4)N)CN in DMF, followed by recrystallization from DMF/MeOH, yields (Et(4)N)(5)[Mo(2)(CN)(11)] x 2DMF x 2MeOH (2). The dinuclear [Mo(2)(CN)(11)](5-) complex featured therein contains two octahedrally coordinated Mo(III) centers spanned by a bridging cyanide ligand. A fit to the magnetic susceptibility data for 2, gives J = -113 cm(-1) and g = 2.33, representing the strongest antiferromagnetic coupling yet observed through a cyanide bridge. Efforts to incorporate these new complexes in magnetic Prussian blue-type solids are ongoing. PMID- 11878954 TI - Yellow-green and red firefly bioluminescence from 5,5-dimethyloxyluciferin. AB - Beetle luciferases (including those of the firefly) use the same luciferin substrate to naturally display light ranging in color from green (lambda(max) similar 530 nm) to red (lambda(max) similar 635 nm). The original mechanism of bioluminescence color determination advanced by White and co-workers was based on the concept that the keto and enol tautomers of the emitter oxyluciferin produce red and green light, respectively. Alternatively, McCapra proposed that color variation is associated with conformations of the keto form of excited-state oxyluciferin. We have prepared the adenylate of D-5,5-dimethylluciferin and shown that it is transformed into the putative emitter 5,5-dimethyloxyluciferin in bioluminescence reactions catalyzed by luciferases from Photinus pyralis and the green-emitting click beetle. 5,5-Dimethyloxyluciferin is constrained to exist in the keto form and fluoresces in the red. However, bioluminescence spectra revealed that green light emission was produced by the firefly enzyme and red light was observed with the click beetle protein. These results, augmented with steady-state kinetic studies, may be taken as the first experimental support for McCapra's mechanism of firefly bioluminescence color or any other proposal that requires only a single keto form of oxyluciferin. PMID- 11878955 TI - Remarkable remote chiral recognition in a reaction mediated by a catalytic antibody. AB - The crystal structures of catalytic antibody D2.3 Fab with the two enantiomers, 7D and 7L, which represent transition state analogues for the hydrolysis of the corresponding esters, 6D and 6L, were determined to better understand remarkable reactivity differences: the L-ester displayed significantly tighter binding (K(M)) and increased catalytic activity (k(cat)) with D2.3, even though the chiral center is 7 bonds distant from the reaction center. Surprisingly, the electron densities of the liganded phosphonates, 7D and 7L, within the D2.3 binding/reaction site were essentially identical, highlighting the subtle influences of protein interactions on chemical behavior. PMID- 11878956 TI - Metastable one-dimensional AgCl(1)-(x)I(x) solid-solution wurzite "tunnel" crystals formed within single-walled carbon nanotubes. AB - High-resolution transmission electron microscopy and spatially resolved electron loss spectroscopy have revealed that a eutectic mixture of AgCl and AgI crystallizes within single walled carbon nanotubes (SWNTs) as metastable AgCl(1 )(x)I(x) 1D solid solution crystals. The incorporated halide crystals form wurzite "tunnel" structures with locally varying Cl:I ratios and reduced Ag coordination. PMID- 11878957 TI - Photochemistry of benzophenone immobilized in a major groove of DNA: formation of thermally reversible interstrand cross-link. AB - We here report a highly site and sequence selective formation of an interstrand cross-link of (BP)U-containing oligomer duplexes. The cross-link was found spontaneously reverted to original oligomers upon heating, providing a new method for the temporary connection of two DNA strands. PMID- 11878958 TI - Electrical contacting of glucose oxidase by surface-reconstitution of the apo protein on a relay-boronic acid-FAD cofactor monolayer. AB - A new methodology for the surface-reconstitution of apo-flavoenzymes on a relay unit (pyrroloquinoline quinone, PQQ) functionalized with a boronic acid ligand as a linker to native FAD was developed. The reconstitution of apo-glucose oxidase (apo-GOx) on the PQQ-FAD monolayer yields an electrically contacted enzyme electrode. PMID- 11878959 TI - Supramolecular stabilization of N(2)H(7)(+). AB - The cation N(2)H(7)(+) has been stabilized in a largely hydrophobic supramolecular environment and characterized in the solid state. The cation is situated in the bowl-shaped cavity of calix[4]arene. All of the hydrogen atoms are clearly discernible owing to high-quality X-ray data as well as lack of disorder and symmetry-imposed ambiguity. It appears that electrostatic interactions play a critical role in stabilizing the structure. PMID- 11878960 TI - Photoswitchable multivalent sugar ligands: synthesis, isomerization, and lectin binding studies of azobenzene-glycopyranoside derivatives. AB - Coating of azobenzene chromophore with multivalent sugar ligands has been accomplished. Such sugar coating allows the study of the isomerization properties of this chromophore in aqueous solutions. The predominantly cis-isomer-containing photostationary state (PS) mixture of these azobenzene derivatives is found to be stable for hours. The rate constants for their isomerization, as well as the Arrhenius activation energies, are determined experimentally. An assessment of the lectin binding properties of the lactoside bearing isomeric azobenzene derivatives, by isothermal calorimetric methods, reveals the existence of an unusual cooperativity in their binding to lectin peanut agglutinin. Thermodynamic parameters evaluated for the trans and the PS mixture are discussed, in detail, for the lactoside bearing bivalent azobenzene derivative. PMID- 11878961 TI - A self-organized 2-dimensional bifunctional structure formed by supramolecular design. AB - The production of a novel two-dimensional bimolecular surface structure using weak noncovalent interactions is demonstrated and observed by scanning tunneling microscopy. This work follows the three-dimensional (3D) ideas of crystal engineering and applies the concepts of supramolecular synthons to molecular systems constrained to 2D by physisorption on a conducting surface. We demonstrate a well-ordered planar structure that self-assembles through the influence of fluorine-phenyl interactions. This study provides a concrete example of the "bottom up" construction of nanostructures and of the systematic design of self-organized layers. To our knowledge, this is the first in a new class of fully 2D materials based both upon weak intermolecular interactions and upon image charges and weak interactions associated with adsorption on metal surfaces. PMID- 11878962 TI - Probing pentacene polymorphs by lattice dynamics calculations. AB - We have performed a lattice dynamics calculation to compute the "inherent structures" of minimum potential energy for pentacene, starting from available X ray data. The calculation shows that two distinct bulk crystalline phases of pentacene exist, with very subtle structural differences but clearly different phonon spectra. The method of crystal growth (from solution or vapor) is not the determining factor for obtaining either structure. PMID- 11878963 TI - Novel zinc (II)-mediated epimerization of 2'-carbonylalkyl-alpha-C glycopyranosides to their beta-anomers. AB - 2'-Aldehydes and 2'-ketones of alpha-C-glycosides, including the gluco-, galacto , and manno- series, were epimerized exclusively to their beta-anomers in good-to excellent yields under basic conditions and in the presence of zinc acetate. The beta-stereoselectivity is independent of the neighboring group at 2-O substitution of sugar substrates. Therefore, this provides a particularly useful method for the preparation of manno-beta-C-glycosides. The epimerization is likely initiated by the formation of Zn-enolate that is stabilized by intramolecular chelation to the pyranose ring-oxygen to form a syn chair-boat structure. Due to the activation generated by the Zn-O coordination, fission of the C1-O bond occurs, leading to opening of the pyranose ring, which is spontaneously followed by a change in conformation. The more stable anti chair boat transition state is favored, and the subsequent hetero-intramolecular Michael addition results in the formation of beta-C-glycoside in a ring-closure step. PMID- 11878964 TI - Cationic scandium methyl complexes supported by a beta-diketiminato ("Nacnac") ligand framework. AB - The base-free dimethyl scandium complex supported by the bulky beta-diketiminato ligand ArNC((t)Bu)CHC((t)Bu)NAr (Ar = 2,6-(i)Pr(2)C(6)H(3), 1) reacts with various equivalencies of the strong organometallic Lewis acid B(C(6)F(5))(3) to give scandium alkyl cations. With 0.5 equiv, a monocationic mu-methyl dimer (2) was observed spectroscopically. Reaction with a further 0.5 equiv of borane gives the monomeric methyl cation 3, which was fully characterized, including via X-ray crystallography. This compound is fluxional on the NMR time scale via a "ligand flip" mechanism. Reaction with another equivalent of borane gives the unique dication 4, which exhibits a static structure on the NMR time scale. Dimethyl compound 1 is a highly active catalyst precursor for ethylene polymerization under borane or MAO-type activation. Activities for this group 3 metal based catalyst approach those observed for group 4 based metallocene systems. PMID- 11878965 TI - Asymmetric azidation-cycloaddition with open-chain peptide-based catalysts. A sequential enantioselective route to triazoles. AB - A family of beta-substituted histidine-containing peptides has been synthesized to probe the effect of noncovalent conformational rigidification on catalyst enantioselectivity. Unambiguous enhancement of enantioselectivity in the conjugate addition of azide to alpha,beta-unsaturated carboxylate derivatives has been achieved, enabling application to a sequential asymmetric azidation/cycloaddition for the synthesis of optically enriched triazoles and triazolines. PMID- 11878966 TI - Stereocontrolled total synthesis of (+)-vinblastine. AB - A stereocontrolled total synthesis of (+)-vinblastine was accomplished, featuring preparations of the two indole units by means of a novel indole synthesis via radical cyclization of thioanilide, and a stereoselective coupling of these units. PMID- 11878967 TI - Designed self-assembly of molecular necklaces. AB - This paper reports an efficient strategy to synthesize molecular necklaces, in which a number of small rings are threaded onto a large ring, utilizing the principles of self-assembly and coordination chemistry. Our strategy involves (1) threading a molecular "bead" with a short "string" to make a pseudorotaxane and then (2) linking the pseudorotaxanes with a metal complex with two cis labile ligands acting as an "angle connector" to form a cyclic product (molecular necklace). A 4- or 3-pyridylmethyl group is attached to each end of 1,4 diaminobutane or 1,5-diaminopentane to produce the short "strings" (C4N4(2+), C4N3(2+), C5N4(2+), and C5N3(2+)), which then react with a cucurbituril (CB) "bead" to form stable pseudorotaxanes (PR44(2+), PR43(2+), PR54(2+), and PR53(2+), respectively). The reaction of the pseudorotaxanes with Pt(en)(NO(3))(2) (en = ethylenediamine) produces a molecular necklace [4]MN, in which three molecular "beads" are threaded on a triangular framework, and/or a molecular necklace [5]MN, in which four molecular "beads" are threaded on a square framework. Under refluxing conditions, the reaction with PR44(2+) or PR54(2+) yields exclusively [4]MN (MN44T or MN54T, respectively), whereas that with PR43(2+) or PR53(2+) produces exclusively [5]MN (MN43S or MN53S, respectively). The products have been characterized by various methods including X-ray crystallography. At lower temperatures, on the other hand, the reaction with PR44(2+) or PR54(2+) affords both [4]MN and [5]MN. The supermolecules reported here are the first series of molecular necklaces obtained as thermodynamic products. The overall structures of the molecular necklaces are strongly influenced by the structures of pseudorotaxane building blocks, which is discussed in detail on the basis of the X-ray crystal structures. The temperature dependence of the product distribution observed in this self-assembly process is also discussed. PMID- 11878968 TI - Hairpin versus extended DNA binding of a substituted beta-alanine linked polyamide. AB - A series of alpha-substituted beta-alanine (beta) linked polyamides (DbaPyPyPy beta*-PyPyPy) were prepared and examined. This resulted in the observation that while most substituents disrupt DNA binding, (R)-alpha-methoxy-beta-alanine (beta((R)-OMe)) maintains strong binding affinity and preferentially adopts a hairpin versus extended binding mode, providing an alternative hairpin linker to gamma-aminobutyric acid (gamma). A generalized variant of a fluorescent intercalator displacement assay conducted on a series of hairpin deoxyoligonucleotides containing a systematically varied A/T-rich binding site size was developed to distinguish between the extended binding of the parent beta alanine 1 (DbaPyPyPy-beta-PyPyPy) and the hairpin binding of 3 (DbaPyPyPy beta((R)-OMe)-PyPyPy). PMID- 11878969 TI - Influence of a terminal formamido group on the sequence recognition of DNA by polyamides. AB - Pyrrole (Py)-imidazole (Im)-containing polyamides bind in the minor groove of DNA and can recognize specific sequences through a stacked antiparallel dimer. It has been proposed that there are two different low energy ways to form the stacked dimer and that these are sensitive to the presence of a terminal formamido group: (i) a fully overlapped stacking mode in which the N-terminal heterocycles of the dimer stack on the amide groups between the two heterocycles at the C-terminal and (ii) a staggered stacking mode in which the N-terminal heterocycles are shifted by approximately one unit in the C-terminal direction (Structure 1997, 5, 1033-1046). Two different DNA sequences will be recognized by the same polyamide stacked in these two different modes. Despite the importance of polyamides as sequence specific DNA recognition agents, these stacking possibilities have not been systematically explored. As part of a program to develop agents that can recognize mismatched base pairs in DNA, a set of four polyamide trimers with and without terminal formamido groups was synthesized, and their interactions with predicted DNA recognition sequences in the two different stacking modes were evaluated. Experimental difficulties in monitoring DNA complex formation with polyamides were overcome by using surface plasmon resonance (SPR) detection of the binding to immobilized DNA hairpin duplexes. Both equilibrium and kinetic results from SPR show that a terminal formamido group has a pronounced effect on the affinity, sequence specificity, and rates of DNA-dimer complex formation. The formamido polyamides bind preferentially in the staggered stacking mode, while the unsubstituted analogues bind in the overlapped mode. Affinities for cognate DNA sequences increase by a factor of around 100 when a terminal formamido is added to a polyamide, and the preferred sequences recognized are also different. Both the association and the dissociation rates are slower for the formamido derivatives, but the effect is larger for the dissociation kinetics. The formamido group thus strongly affects the interaction of polyamides with DNA and changes the preferred DNA sequences that are recognized by a specific polyamide stacked dimer. PMID- 11878971 TI - Reactivity of amino acids in nitrosation reactions and its relation to the alkylating potential of their products. AB - Nitrosation reactions of amino acids with an -NH(2) group [namely, six alpha amino acids (glycine, alanine, alpha-aminobutyric acid, alpha-aminoisobutyric acid, valine, and norvaline); two beta-amino acids (beta-alanine and beta aminobutyric acid), and one gamma-amino acid (gamma-aminobutyric acid)] were studied. Nitrosation was carried out in aqueous acid media, mimicking the conditions of the stomach lumen. The rate equation was r = k(3)(exp)[amino acid][nitrite](2), with a maximum k(3)(exp) value in the 2.3-2.7 pH range. The existence of an isokinetic relationship supports the argument that all the reactions share a common mechanism. A nitrosation mechanism is proposed, and the following conclusions are drawn: (i) Nitrosation reactions of amino acids with a primary amino group in acid media occur with dinitrogen trioxide as the main nitrosating agent. The finding that the nitrosation rate is proportional to the square of the nitrite concentration suggests that the yield of nitrosation products in the stomach would increase sharply with higher nitrate/nitrite intakes. (ii) Stomach hypochlorhydria could be a potential enhancer of in vivo amino acid nitrosation. (iii) The reactivity (k(3)()(exp)) [alpha-amino acids > beta-amino acids > gamma-amino acids] is the same as that found in a previous work for the alkylating potential of lactones formed from nitrosation products of the same amino acids. This implies that the nitrosation reactions of the most common natural amino acids are the most efficient precursors of the most powerful alkylating agents. (iv) The order of magnitude (10(7)-10(8) M(-1) s(-1)) of the bimolecular rate constants of nitrosation shows that such reactions occur through an encounter process. PMID- 11878970 TI - alpha-L-ribo-configured locked nucleic acid (alpha-L-LNA): synthesis and properties. AB - The syntheses of monomeric nucleosides and 3'-O-phosphoramidite building blocks en route to alpha-L-ribo-configured locked nucleic acids (alpha-L-LNA), composed entirely of alpha-L-LNA monomers (alpha-L-ribo configuration) or of a mixture of alpha-L-LNA and DNA monomers (beta-D-ribo configuration), are described and the alpha-L-LNA oligomers are studied. Bicyclic 5-methylcytosin-1-yl and adenine-9-yl nucleoside derivatives have been prepared and the phosphoramidite approach has been used for the automated oligomerization leading to alpha-L-LNA oligomers. Binding studies revealed very efficient recognition of single-stranded DNA and RNA target oligonucleotide strands. Thus, stereoirregular alpha-L-LNA 11-mers containing a mixture of alpha-L-LNA monomers and DNA monomers ("mix-mer alpha-L LNA") were shown to display DeltaT(m) values of +1 to +3 degrees C per modification toward DNA and +4 to +5 degrees C toward RNA when compared with the corresponding unmodified DNA x DNA and DNA x RNA reference duplexes. The corresponding DeltaT(m) values per modification for the stereoregular fully modified alpha-L-LNA were determined to be +4 degrees C (against DNA) and +5 degrees C (against RNA). 11-Mer alpha-L-LNAs (mix-mer alpha- L-LNA or fully modified alpha- L-LNA) were shown in vitro to be significantly stabilized toward 3'-exonucleolytic degradation. A duplex formed between RNA and either mix-mer alpha-L-LNA or fully modified alpha-L-LNA induced in vitro Escherichia coli RNase H-mediated cleavage, albeit very slow, of the RNA targets at high enzyme concentrations. PMID- 11878972 TI - Total synthesis of the CP-molecules (CP-263,114 and CP-225,917, phomoidrides B and A). 1. Racemic and asymmetric synthesis of bicyclo[4.3.1] key building blocks. AB - A brief introduction into the chemistry of the CP-molecules is followed by first generation synthetic sequences toward key building blocks for their total synthesis. Processes for both racemic and enantiomerically enriched bicyclo[4.3.1] ketone 6 or its equivalent are described, and the absolute stereochemistries of the optically enriched intermediates are determined. The efficient route developed to racemic 6 and the ready access to both enantiomers of key building blocks provided the opportunity for the total synthesis of the CP molecules and determination of their absolute stereochemistry. PMID- 11878973 TI - Total synthesis of the CP-molecules (CP-263,114 and CP-225,917, phomoidrides B and A). 2. Model studies for the construction of key structural elements and first-generation strategy. AB - Crucial model synthetic and mechanistic studies directed toward the development of methodology for the construction of the maleic anhydride moiety of the CP molecules are described. Studies directed toward the stereoselective attachment of the upper side chain, culminating in the discovery of long-range stereochemical control, are also discussed. In addition, a first-generation strategy toward the CP-molecules, establishing key intermediate 5 as a "beachhead" from which all future operations would diverge, is also presented. Although this first-generation strategy failed to yield the target molecules, the endeavor laid the important groundwork for the next-generation drives toward the CP-molecules. PMID- 11878974 TI - Total synthesis of the CP-molecules (CP-263,114 and CP-225,917, phomoidrides B and A). 3. Completion and synthesis of advanced analogues. AB - The completion of the total syntheses of the CP-molecules is reported. Several strategies and tactics, including the use of amide-based protecting groups for the homologated C-29 carboxylic acid and the use of an internal pyran protecting group scheme, are discussed. The endeavors leading to the design of new methods for the homologation of hindered aldehydes and to the isolation of a polycyclic byproduct (23), which inspired the development of a new series of reactions based on iodine(V) reagents, are described. In addition, the discovery and development of the LiOH-mediated conversion of CP-263,114 (1) to CP-225,917 (2) is described, and a mechanistic rationale is presented. Finally, a synthetic route to complex analogues of the CP-molecules harboring a maleimide moiety in place of the maleic anhydride is presented. PMID- 11878975 TI - Iodine(V) reagents in organic synthesis. Part 1. Synthesis of polycyclic heterocycles via Dess-Martin periodinane-mediated cascade cyclization: generality, scope, and mechanism of the reaction. AB - The scope, generality, and mechanism of the Dess-Martin periodinane-mediated cyclization reaction of unsaturated anilides discovered during the total synthesis of the CP-molecules (phomoidrides A and B) are delineated. A plethora of heterocyclic compounds are accessible by employing gamma,delta-unsaturated amides (derived from anilines and carboxylic acids), urethanes, or ureas (derived from isocyanates and allylic alcohols and amines) as substrates. Optimization of the reaction led to room-temperature conditions, while isotope labeling studies allowed a mechanistic rationale for this cascade reaction. PMID- 11878976 TI - Iodine(V) reagents in organic synthesis. Part 2. Access to complex molecular architectures via Dess-Martin periodinane-generated o-imidoquinones. AB - o-Imidoquinones, a rather rare class of compounds, are prepared from anilides by the action of Dess-Martin periodinane (DMP) and water. Their chemistry has been extensively investigated and found to lead to p-quinones and polycyclic systems of diverse molecular architectures. Applications of this methodology to the total synthesis of the naturally occurring compounds, epoxyquinomycin B and BE-10988, are described. Finally, another rare chemical entity, the ketohydroxyamide moiety, has been accessed through this DMP-based synthetic technology, and its reactivity has been studied. Among its most useful reactions is a set of cascade heterocyclic annulations leading to a variety of polycyclic systems of possible biological relevance. PMID- 11878977 TI - Iodine(V) reagents in organic synthesis. Part 3. New routes to heterocyclic compounds via o-iodoxybenzoic acid-mediated cyclizations: generality, scope, and mechanism. AB - The discovery and development of the o-iodoxybenzoic acid (IBX) reaction with certain unsaturated N-aryl amides (anilides) to form heterocycles are described. The application of the method to the synthesis of delta-lactams, cyclic urethanes, hydroxy amines, and amino sugars among other important building blocks and intermediates is detailed. In addition to the generality and scope of this cyclization reaction, this article describes a number of mechanistic investigations suggesting a single electron transfer from the anilide functionality to IBX and implicating a radical-based mechanism for the reaction. PMID- 11878978 TI - Iodine(V) reagents in organic synthesis. Part 4. o-Iodoxybenzoic acid as a chemospecific tool for single electron transfer-based oxidation processes. AB - o-Iodoxybenzoic acid (IBX), a readily available hypervalent iodine(V) reagent, was found to be highly effective in carrying out oxidations adjacent to carbonyl functionalities (to form alpha,beta-unsaturated carbonyl compounds) and at benzylic and related carbon centers (to form conjugated aromatic carbonyl systems). Mechanistic investigations led to the conclusion that these new reactions are initiated by single electron transfer (SET) from the substrate to IBX to form a radical cation which reacts further to give the final products. Fine-tuning of the reaction conditions allowed remarkably selective transformations within multifunctional substrates, elevating the status of this reagent to that of a highly useful and chemoselective oxidant. PMID- 11878979 TI - Hydroxide as general base in the saponification of ethyl acetate. AB - The second-order rate constant for the saponification of ethyl acetate at 30.0 degrees C in H(2)O/D(2)O mixtures of deuterium atom fraction n (a proton inventory experiment) obeys the relation k(2)(n) = 0.122 s(-1) M(-1) (1 - n + 1.2n) (1 - n + 0.48n)/(1 - n + 1.4n) (1 - n + 0.68n)(3). This result is interpreted as a process where formation of the tetrahedral intermediate is the rate-determining step and the transition-state complex is formed via nucleophilic interaction of a water molecule with general-base assistance from hydroxide ion, opposite to the direct nucleophilic collision commonly accepted. This mechanistic picture agrees with previous heavy-atom kinetic isotope effect data of Marlier on the alkaline hydrolysis of methyl formate. PMID- 11878980 TI - Confirmation of the connectivity of 4,8,12,16,20-pentamethylpentacosylphoshoryl beta-D-mannopyranoside, an unusual beta-mannosyl phosphoisoprenoid from Mycobacterium avium, through synthesis. AB - The synthesis of the title glycolipid is reported. Comparison of the electrospray and high-energy collision-induced dissociation mass spectra of the synthetic material with those reported for the isolate confirm the structure of this unusual antigenic substance with its modified isoprenoid chain. PMID- 11878982 TI - Nickel(II)-molybdenum(III)-cyanide clusters: synthesis and magnetic behavior of species incorporating [(Me(3)tacn)Mo(CN)(3)]. AB - The substitution of Mo(III) for Cr(III) in metal-cyanide clusters is demonstrated as an effective means of increasing the strength of the magnetic exchange coupling and introducing magnetic anisotropy. Synthesis of the octahedral complex [(Me(3)tacn)Mo(CN)(3)] (Me(3)tacn = N,N',N"-trimethyl-1,4,7-triazacyclononane) is accomplished with the addition of precisely 3 equiv of LiCN to a solution of [(Me(3)tacn)Mo(CF(3)SO(3))(3)] in DMF. An excess of LiCN prompts formation of a seven-coordinate complex, [(Me(3)tacn)Mo(CN)(4)](1)(-), whereas less LiCN produces multinuclear species such as [(Me(3)tacn)(2)Mo(2)(CN)(5)](1+). In close parallel to reactions previously performed with [(Me(3)tacn)Cr(CN)(3)], assembly reactions between [(Me(3)tacn)Mo(CN)(3)] and [Ni(H(2)O)(6)](2+) or [(cyclam)Ni(H(2)O)(2)](2+) (cyclam = 1,4,8,11-tetraazacyclotetradecane) afford face-centered cubic [(Me(3)tacn)(8)Mo(8)Ni(6)(CN)(24)](12+) and linear [(Me(3)tacn)(2)(cyclam)NiMo(2)(CN)(6)](2+) clusters, respectively. Generation of the former involves a thermally induced cyanide linkage isomerization, which rapidly leads to a low-spin form of the cluster containing diamagnetic Ni(II) centers. The cyclic voltammagram of this species in DMF reveals a sequence of six successive reduction waves spaced approximately 130 mV apart, suggesting class II mixed-valence behavior upon reduction. The magnetic properties of the aforementioned linear cluster are consistent with the expected ferromagnetic coupling and an S = 4 ground state, but otherwise vary slightly with the specific conformation adopted (as influenced by the packing of associated counteranions and solvate molecules in the crystal). Magnetization data indicate an axial zero field splitting parameter with a magnitude falling in the range [D] = 0.44-0.72 cm(-1), and fits to the magnetic susceptibility data yield exchange coupling constants in the range J = 17.0-17.6 cm(-1). These values represent significant increases over those displayed by the analogous Cr(III)-containing cluster. When perchlorate is used as a counteranion, [(Me(3)tacn)(2)(cyclam)NiMo(2)(CN)(6)](2+) crystallizes from water in a dimeric form with pairs of the linear clusters directly linked via hydrogen bonding. In this case, fitting the magnetic susceptibility data requires use of two coupling constants: one intramolecular with J = 14.9 cm(-1) and another intermolecular with J' = -1.9 cm(-1). Reacting [(Me(3)tacn)Mo(CN)(3)] with a large excess of [(cyclam)Ni(H(2)O)(2)](2+) produces a [(Me(3)tacn)(2)(cyclam)(3)(H(2)O)(2)Ni(3)Mo(2)(CN)(6)](6+) cluster possessing a zigzag structure that is a simple extension of the linear cluster geometry. Its magnetic behavior is consistent with weaker ferromagnetic coupling and an S = 6 ground state. Similar reactions employing an equimolar ratio of reactants afford related one-dimensional chains of formula [(Me(3)tacn)(cyclam)NiMo(CN)(3)](2+). Once again, the ensuing structure depends on the associated counteranions, and the magnetic behavior indicates ferromagnetic coupling. It is hoped that substitutions of the type exemplified here will be of utility in the design of new single-molecule magnets. PMID- 11878981 TI - Ion channel formation from a calix[4]arene amide that binds HCl. AB - The ion transport activity of calix[4]arene tetrabutylamide 1,3-alt 2 was studied in liposomes, planar lipid bilayers, and HEK-293 cells. These experiments, when considered together with (1)H NMR and X-ray crystallography data, indicate that calix[4]arene tetrabutylamide 2 (1) forms ion channels in bilayer membranes, (2) mediates ion transport across cell membranes at positive holding potential, (3) alters the pH inside liposomes experiencing a Cl(-) gradient, and (4) shows a significant Cl(-)/SO(4)(2)(-) transport selectivity. An analogue, calix[4]arene tetramethylamide 1, self-assembles in the presence of HCl to generate solid-state structures with chloride-filled and water-filled channels. Structureminus signactivity studies indicate that the hydrophobicity, amide substitution, and macrocyclic framework of the calixarene are essential for HCl binding and transport. Calix[4]arene tetrabutylamide 2 is a rare example of an anion dependent, synthetic ion channel. PMID- 11878983 TI - Stabilization of inorganic nanocrystals by organic dendrons. AB - A series of hydrophilic organic dendron ligands was designed and synthesized for stabilizing high-quality semiconductor and noble metal nanocrystals. The focal point of the dendron ligands is chosen to be a thiol group which is a universal coordinating site for compound semiconductor and noble metal nanocrystals. The methods for binding these dendron ligands onto the surface of the nanocrystals are simple and straightforward. The thin, about 1-2 nm, but closely packed and tangled ligand shell provides sufficient stability for the "dendron-protected nanocrystals" to withstand the rigors of the coupling chemistry and the standard separation/purification techniques. The chemistry presented can be immediately applied for the development of a new generation of biomedical labeling reagents based on high-quality semiconductor nanocrystals. It also provides an alternative path to apply noble metal nanocrystals for developing sensitive detection schemes for chemical and biochemical purposes. The concept may further provide an optimal solution for many other problems encountered in nanocrystal-related research and development, for which the stability of the nanocrystals is a critical issue. Furthermore, the experimental results confirmed that the photochemical stability of colloidal semiconductor and noble metal nanocrystals is the key for developing reliable and reproducible processing chemistry for these nanocrystals. PMID- 11878984 TI - Physical organic chemistry of transition metal carbene complexes. 24. Thermodynamic and kinetic acidities of phenyl-substituted (benzylmethoxycarbene)pentacarbonylchromium(0) complexes. Is there a transition state imbalance? AB - A kinetic study of the reversible deprotonation of phenyl-substituted (benzylmethoxycarbene)pentacarbonylchromium(0) complexes by OH(-) and by a series of primary aliphatic and a series of secondary alicyclic amines in 50% MeCN-50% water (v/v) at 25 degrees C is reported. Bronsted alpha(CH) values (dependence on carbene complex acidity) and beta(B) values (dependence on amine basicity) were determined. According to current notions about proton transfers involving carbon acids activated by pi-acceptors, alpha(CH) was expected to substantially exceed beta(B), the result of transition-state imbalances that are characteristic of such reactions. However we find that alpha(CH) and beta(B) have essentially the same values, which are close to 0.5. It is shown that these findings do not indicate the absence of an imbalance but rather suggest that the manifestation of the imbalance is masked by the pi-donor effect (3H-Z <--> 3H-Z(+/-)) of the methoxy group. PMID- 11878985 TI - Gram-scale synthesis of monodisperse gold colloids by the solvated metal atom dispersion method and digestive ripening and their organization into two- and three-dimensional structures. AB - We describe a synthetic procedure for preparation of large quantities of monodisperse thiol-stabilized gold colloids in toluene solution. The method is based on the solvated metal atom dispersion technique (SMAD), which is very suitable for preparation of large amounts of metal colloidal solutions, as well as of metal sulfide, metal oxide, and other types of dispersed compounds in different solvents. A combination of two different solvents like acetone and toluene is used for the preparation of the gold colloids. The necessity of initially carrying out the SMAD reaction in acetone comes from its high degree of solvation of gold particles. Acetone acts as a preliminary stabilizing agent. After its removal from the system, the particles are stabilized by dodecanethiol molecules, which enable their very good dispersion in toluene solution. A digestive ripening procedure is carried out with the gold-toluene colloid, and for this purpose pure toluene as solvent is necessary. This has a dramatic effect on the narrowing of particle size distribution and almost monodisperse colloids are obtained (some discussion of the probable mechanism of this remarkable digestive ripening step is given). These colloidal solutions have a great tendency to organize in two- and three-dimensional structures (nanocrystal superlattices, NCSs). We believe that this procedure provides a real opportunity to synthesize large amounts of gold nanocrystals as well as NCSs. PMID- 11878986 TI - Size-dependent melting of silica-encapsulated gold nanoparticles. AB - We report on the size dependence of the melting temperature of silica encapsulated gold nanoparticles. The melting point was determined using differential thermal analysis (DTA) coupled to thermal gravimetric analysis (TGA) techniques. The small gold particles, with sizes ranging from 1.5 to 20 nm, were synthesized using radiolytic and chemical reduction procedures and then coated with porous silica shells to isolate the particles from one another. The resulting silica-encapsulated gold particles show clear melting endotherms in the DTA scan with no accompanying weight loss of the material in the TGA examination. The silica shell acts as a nanocrucible for the melting gold with little effect on the melting temperature itself, even though the analytical procedure destroys the particles once they melt. Phenomenological thermodynamic predictions of the size dependence of the melting point of gold agree with the experimental observation. Implications of these observations to the self-diffusion coefficient of gold in the nanoparticles are discussed, especially as they relate to the spontaneous alloying of core-shell bimetallic particles. PMID- 11878987 TI - EPR study of substrate binding to the Mn(II) active site of the bacterial antibiotic resistance enzyme FosA: a better way to examine Mn(II). AB - FosA is a manganese metalloglutathione transferase that confers resistance to the broad-spectrum antibiotic fosfomycin, (1R,2S)-epoxypropylphosphonic acid. The reaction catalyzed by FosA involves the attack by glutathione on fosfomycin to yield the product 1-(S-glutathionyl)-2-hydroxypropylphosphonic acid. The enzyme is a dimer of 16 kDa subunits, each of which harbors one mononuclear Mn(II) site. The coordination environment of the Mn(II) in the FosA x Mn(2+) complex is composed of a glutamate and two histidine ligands and three water molecules. Here we report EPR spectroscopic studies on FosA, in which EPR spectra were obtained at 35 GHz and 2 K using dispersion-detection rapid-passage techniques. This approach provides an absorption envelope line shape, in contrast to the conventional (slow-passage) derivative line shape, and is a more reliable way to collect spectra from Mn(II) centers with large zero-field splitting. We obtain excellent spectra of FosA bound with substrate, substrate analogue phosphate ion, and product, whereas these states cannot be studied by X-band, slow-passage methods. Simulation of the EPR spectra shows that binding of substrate or analogue causes a profound change in the electronic parameters of the Mn(II) ion. The axial zero-field splitting changes from [D] = 0.06 cm(-1) for substrate-free enzyme to 0.23 cm(-1) for fosfomycin-bound enzyme, 0.28 (1) cm(-1) for FosA with phosphate, and 0.27 (1) cm(-1) with product. Such a large zero-field splitting is uncommon for Mn(II). A simple ligand field analysis of this change indicates that binding of the phosphonate/phosphate group of substrate or analogue changes the electronic energy levels of the Mn(II) 3d orbitals by several thousand cm(-1), indicative of a significant change in the Mn(II) coordination sphere. Comparison with related enzymes (glyoxalase I and MnSOD) suggests that the change in the coordination environment on substrate binding may correspond to loss of the glutamate ligand. PMID- 11878988 TI - Remarkable variations in the luminescence of frozen solutions of [Au[C(NHMe)(2)](2)](PF(6)) x 0.5(acetone). Structural and spectroscopic studies of the effects of anions and solvents on Gold(I) carbene complexes. AB - The unusual luminescence behavior of the two-coordinate gold(I) carbene complex, [Au[C(NHMe)(2)](2)](PF(6)) x 0.5(acetone), is reported. Upon freezing in a liquid N(2) bath, the colorless, nonluminescent solutions of [Au[C(NHMe)(2)](2)](PF(6)) x 0.5(acetone) become intensely luminescent. Strikingly, the colors of the emission differ in different solvents and appear only after the solvent has frozen. Solid [Au[C(NHMe)(2)](2)](PF(6)) x 0.5(acetone) is also luminescent, and the luminescence is attributed to the formation of extended chains of gold(I) centers that are connected through aurophilic attractions. Crystallographic studies of [Au[C(NHMe)(2)](2)](PF(6)) x 0.5(acetone) and [Au[C(NHMe)(2)](2)](BF(4)), which is also luminescent, reveal that both involve extended chains of cations and that the anions are hydrogen bonded to the cations through cation N-H groups. However, these chains differ in the Au...Au separations in each and in the carbene ligand orientations. In contrast, [Au[C(NMe(2))(NHMe)](2)](PF(6)) forms a colorless, nonluminescent solid, and in that solid there are no Au...Au interactions, a factor which supports the contention that aggregated species are responsible for the luminescence of [Au[C(NHMe)(2)](2)](PF(6)) x 0.5(acetone) in the solid state and in frozen solutions. PMID- 11878989 TI - Identification of ligand binding sites on proteins using a multi-scale approach. AB - Identification of a ligand binding site on a protein is pivotal to drug discovery. To date, no reliable and computationally feasible general approach to this problem has been published. Here we present an automated efficient method for determining binding sites on proteins for potential ligands without any a priori knowledge. Our method is based upon the multiscale concept where we deal with a hierarchy of models generated using a k-means clustering algorithm for the potential ligand. This is done in a simple approach whereby a potential ligand is represented by a growing number of feature points. At each increasing level of detail, a pruning of potential binding site is performed. A nonbonding energy function is used to score the interactions between molecules at each step. The technique was successfully employed to seven protein-ligand complexes. In the current paper we show that the algorithm considerably reduces the computational effort required to solve this problem. This approach offers real opportunities for exploiting the large number of structures that will evolve from structural genomics. PMID- 11878990 TI - Solid-state NMR and calorimetry of structural waters in helical peptides. AB - The peptide hydrates Gly-Gly-Val x 2H(2)O (GGV) and Gly-Ala-Leu x 3H(2)O (GAL) are known to adopt alpha-helical configurations containing waters of hydration in which each water is H-bonded to three or four peptide groups. Herein we report a thermodynamic and solid-state NMR ((2)H and (17)O) study of these peptides. From TGA and DSC, the average enthalpy per H-bond is 15 kJ/mol. The dynamics and average orientation of the hydrate are studied by powder and single-crystal (2)H NMR. Whereas waters that are shown by the X-ray structure to be coordinated by four hydrogen bonds do not yield observable (2)H NMR signals at room temperature, two of the three triply coordinated waters yield residual (2)H quadrupole coupling tensors characteristic of rapid 180 degrees flip motions and the orientation of the residual tensor is that expected from the X-ray structure derived H-bonding pattern. At -65 degrees C, the flip motions of triply coordinated water in GGV slow into the (2)H NMR intermediate exchange regime whereas the tetrahedrally coordinated water approaches the slow-exchange limit and yields an observable NMR signal. Extensive isotope exchange between water vapor and crystalline GGV establishes the presence of additional hydrate dynamics and solid-state proton transfer along a chain of water-bridged protonated alpha amino groups. PMID- 11878991 TI - Conformational changes of 2,11-dithia[3.3]metacyclophane. A new look using VT NMR and calculation. AB - The conformational changes of 2,11-dithia[3.3]metacyclophane are reexamined utilizing VT NMR data and calculations (ab initio, semiempirical, density functional, and molecular mechanics) to show that the syn to syn' inversion that occurs with exchange of the benzylic hydrogens proceeds most easily through bridge wobbles of the anti isomers and that the critical barrier is the conversion of the syn - chair - chair isomer to the anti-chair-chair isomer, barrier (found) = 9.3-9.6 kcal/mol, barrier (calc) = 10.3 kcal/mol, such that when this conversion is slow on the NMR time scale, the benzylic hydrogens no longer exchange, and the syn-chair-chair isomer becomes frozen. The syn-boat chair isomer, however, can continue to invert to the anti-boat-chair isomer until lower temperatures, and thus, the benzylic hydrogens continue to exchange for this isomer. Thus, while bridge wobbles of the syn isomers have the largest barriers, bridge wobbles of the anti isomers have the smallest barriers, and so the barriers of the syn-to-anti conversions play a much greater role than previously determined. PMID- 11878992 TI - An alternative method for pucker determination in carbohydrates from residual dipolar couplings: a solution NMR study of the fructofuranosyl ring of sucrose. AB - A simple solution NMR method is presented for pucker determination of five membered rings using only residual dipolar couplings obtained in a single liquid crystalline medium, DMPC/DHPC bicelles (DMPC = dimyristoylphosphatidylcholine; DHPC = dihexanoylphosphatidylcholine). The method was applied to determine the pucker of the fructofuranosyl ring of sucrose. The results indicate a fructofuranosyl pucker phase in the 20 degrees - 70 degrees range. The pucker phases are in agreement with those from previous NMR and optical spectroscopic studies and, importantly, do not rely on empirically parametrized Karplus curves. Furthermore, the analysis implies more than one stable pucker phase and rapid ring interconversion in this range. The present results suggest that using residual dipolar couplings alone can reveal multiple conformations present in solution. Hence, when a sufficient number of residual dipolar coupling constants is measured, the outcome is a robust, reliable, and independent route for determining carbohydrate and nucleic acid structure by NMR spectroscopy. PMID- 11878993 TI - Stability and aromaticity of the cyclopenta-fused pyrene congeners. AB - The aromaticity of all possible cyclopenta-fused pyrene congeners has been investigated at various levels of theory. On the basis of the calculated resonance energies and magnetic properties (delta(1)H data, magnetic susceptibility anisotropies, and NICS values), the overall aromaticity of these compounds is found to decrease gradually with increasing number of externally fused five-membered rings. The relatively small differences (<5 kcal/mol) in thermodynamic stability of the isomeric dicyclopentapyrenes (E(tot): dicyclopenta[cd,fg]- > dicyclopenta[cd,jk]- > dicyclopenta[cd,mn]pyrene), which differs from the aromaticity order based on the magnetic criteria (dicyclopenta[cd,mn]- > dicyclopenta[cd,fg]- > dicyclopenta[cd,jk]pyrene), is shown by model calculations to be dominated by sigma-strain imposed on the pyrene skeleton by sequential cyclopenta-fusion. This is supported by the computed homodesmotic reaction energies and aromatic stabilization energy (ASE(isom)) from isodesmic aromatic-nonaromatic isomerization, and by the model calculations on "distorted" cyclopenta[cd]pyrenes. The elusive tetracyclopenta[cd,fg,jk,mn]pyrene is computed to be bowl-shaped; its corresponding planar geometry is the transition state for bowl-bowl interconversion. PMID- 11878994 TI - Methyl substituent effects in [H(n)X...XH(n)](+) three-electron-bonded radical cations (X = F, O, N, Cl, S, P; n = 1 - 3). An ab initio theoretical study. AB - The effects of methyl substitution on the geometries and bonding energies of a systematic series of three-electron-bonded radical cations of the type [H(n)X...XH(n)](+), covering all possible symmetrical three-electron bonds that may take place between atoms of the second and third rows of the periodic table, have been investigated at the level of Moller-Plesset perturbation theory. Methyl substitution leads to significant weakening and lengthening of the X...X bond when X is a second-row atom. The effects increase with the number of substitutions and are more and more important in the series X = N, O, F. By contrast, methyl substitution leaves the bonding energies between third-row atoms practically unchanged but leads to a surprising bond shortening in the S...S and P...P cases. These seemingly contradictory effects are rationalized through a qualitative analysis based on an elementary molecular orbital description of three-electron bonding. PMID- 11878995 TI - Proton diffusion at phospholipid assemblies. AB - A new scanning electrochemical microscopy proton feedback method has been developed for investigating lateral proton diffusion at phospholipid assemblies: specifically Langmuir monolayers at the water/air interface. In this approach, a base is electrogenerated by the reduction of a weak acid (producing hydrogen) at a "submarine" ultramicroelectrode (UME) placed in the aqueous subphase of a Langmuir trough close to a monolayer. The electrogenerated base diffuses to and titrates monolayer-bound protons and is converted back to its initial form, so enhancing the current response at the UME. Local deprotonation of the monolayer creates a concentration gradient for lateral proton diffusion. A numerical model has been developed, taking into account the potential-dependent association/dissociation constant of the interfacial acid groups. A comparison is made of monolayers comprising either acidic DL-alpha-phosphatidyl-L-serine, dipalmitoyl (DPPS) or zwitterionic L-alpha-phosphatidylcholine, dipalmitoyl (DPPC) monolayers at a range of surface pressures. It is demonstrated that lateral proton fluxes at DPPS are significant, but the lateral proton diffusion coefficient is lower than in bulk solution. In contrast, lateral proton diffusion cannot be detected at DPPC, suggesting that the acid/base character of the phospholipid is important in determining the magnitude of interfacial proton fluxes. PMID- 11878996 TI - Enantiospecific desorption of chiral compounds from chiral Cu(643) and achiral Cu(111) surfaces. AB - Temperature-programmed desorption (TPD) experiments have been conducted to investigate enantiospecific desorption from chiral single-crystal surfaces. The (643) and (six four three) planes of face-centered cubic metals such as Cu have kinked and stepped structures which are nonsuperimposable mirror images of one another and therefore are chiral. These chiral surfaces are denoted Cu(643)(R) and Cu(643)(S). We have observed that the desorption energies of (R)-3 methylcyclohexanone and (R)- and (S)-propylene oxides from the Cu(643)(R) and Cu(643)(S) surfaces depend on the relative handedness of the adsorbate/substrate combination. Since the (643) surface is comprised of terraces with local (111) orientation which are separated by kinked monatomic steps, it is instructive to perform TPD experiments with these chiral compounds on the achiral Cu(111) surface. These experiments have given some insight into the adsorption sites for the chiral molecules on the Cu(643) surfaces. There are several high-temperature features in the TPD spectra of the chiral compounds that only appear in the spectra from the (643) surfaces and thus are attributed to molecules adsorbed at or near the kinked steps. In addition there are lower temperature desorption features observed on the Cu(643) surfaces which occur in the same temperature range as desorption features observed on the Cu(111) surface. These features observed on the (643) surfaces are attributed to desorption from the flat (111) terraces. PMID- 11878997 TI - Detection of adulteration in cooked meat products by mid-infrared spectroscopy. AB - Mid-infrared spectroscopy was used to discriminate between pure beef and beef containing 20% w/w of a range of potential adulterants (heart, tripe, kidney, and liver). Spectra were acquired from raw samples and from samples cooked using two different cooking regimes. Chemometric methods (principal component analysis, partial least squares regression, and linear discriminant analysis) applied to the spectra showed that discrimination between the pure and adulterated sample types was possible, irrespective of cooking regime. The cross-validated classification success rate obtained was approximately 97%. Discrimination between all five sample types (pure beef and beef containing one of each of the four adulterants) at each level of cook was also possible, but became more difficult as the cooking level increased. PMID- 11878998 TI - NMR-based screening method for transglutaminases: rapid analysis of their substrate specificities and reaction rates. AB - Incorporation of inter- or intramolecular covalent cross-links into food proteins with microbial transglutaminase (MTG) improves the physical and textural properties of many food proteins such as tofu, boiled fish paste, and sausage. Other transglutaminases (TGases) are expected to be used in the same way, and also to extend the scope of industrial applications to materials, drugs, and so on. The TGases have great diversity, not only in amino acid sequence and size, but also in their substrate specificities and catalytic activities, and therefore, it is quite difficult to estimate their reactivity. We have developed an NMR-based method using the enzymatic labeling technique (ELT) for simultaneous analysis of the substrate specificities and reaction rates of TGases. It is quite useful for comparing the existing TGases and for screening new TGases or TGases variants. This method has shown that MTG is superior for industrial use because of its lower substrate specificity compared with those of guinea pig liver transglutaminase (GTG) and red sea bream liver transglutaminase (FTG). We have also found that an MTG variant lacking an N-terminal aspartic acid residue has higher activity than that of the native enzyme. PMID- 11878999 TI - Phase transitions in the starch-water system studied by adiabatic scanning calorimetry. AB - We report high-precision measurements of phase transitions in the starch-water system by using for the first time adiabatic scanning calorimetry (ASC). Potato starch and nixtamalized corn flours were studied by this technique as a function of the moisture content. We calculated the percentage of gelatinized starch granules, as a function of the temperature, for both flours. For bi-phasic behavior in potato starch, at intermediate water contents, we propose an explanation based on previous hypotheses and on the experimental behavior as deduced from ASC measurements. ASC results suggest, in the case of nixtamalized corn flour, that uniform moisture is present in corn starch granules for all the considered moistures. A shift to higher temperatures of the peak temperature is observed when comparing the c(p)(T) curves with the classical DSC endotherms as measured for this flour. For both flours we estimated from the ASC results the melting temperatures of the starch granules, at zero moisture content, by using Flory's equation. PMID- 11879000 TI - Comparison of extraction methods for marker compounds in the essential oil of lemon grass by GC. AB - A gas chromatography flame ionization detection method for the quantification of bioactive marker compounds (neral, geranial, geraniol, limonene, citronellal, and beta-myrcene) in the essential oil of Cymbopogon citratus (lemon grass) was developed. Four procedures for the extraction of essential oils from C. citratus were compared including solvent extraction, steam distillation extraction, accelerated solvent extraction, and supercritical fluid extraction. Solvent extraction by sonication with nonpolar solvents showed comparable results to the steam distillation method. Several commercial products prepared from C. citratus and Cymbopogon flexuosus were analyzed and compared. PMID- 11879001 TI - A simple method for the determination of trace levels of alkylphenolic compounds in fish tissue using pressurized fluid extraction, solid phase cleanup, and high performance liquid chromatography fluorescence detection. AB - A simple automated extraction method for the determination of alkylphenolic compounds in fish tissue is reported. Pressurized fluid extraction is used to extract ground fish tissue, and the resulting extract is purified on aminopropyl silica (APS) extraction cartridges. With no further sample preparation, nonylphenol (NP) and its ethoxylates, up to nonylphenol pentaethoxylate, are quantitated using normal phase (APS Hypersil) high-performance liquid chromatography with fluorescence detection. The major advantage of this technique is elimination of the conventional gel permeation cleanup step, a lengthy procedure designed to remove fish lipids. Spiked recoveries with lake trout averaged 85% for the six NP and NP ethoxylates that were investigated. Tissue concentrations of NP and NP ethoxylates determined in fish from various locations of the Great Lakes region ranged from 18 to 2075 ng/g, wet weight. PMID- 11879002 TI - Study on the supramolecular interaction of curcumin and beta-cyclodextrin by spectrophotometry and its analytical application. AB - The supramolecular interaction of curcumin and beta-cyclodextrin (beta-CD) has been studied by spectrophotometry. The mechanism of the inclusion was studied and discussed based on the variations of pK(a), absorption intensity, and infrared spectrograms. The results show that beta-CD reacts with curcumin to form a 2:1 host-guest complex with an apparent formation constant of 5.53 x 10(5) mol(-2) x L2. Based on the enhancement of the absorbance of curcumin produced through complex formation, a spectrophotometric method for the determination of curcumin in bulk aqueous solution in the presence of beta-CD was developed. The linear relationship between the absorbance and curcumin concentration was obtained in the range of 0-15 microg/mL, with a correlation coefficient (r) of 0.9991. The detection limit was 0.076 microg/mL. The proposed method was used to determine the curcumin in curry and mustard with satisfactory results. PMID- 11879003 TI - Assessment of the floral origin of honey by SDS-page immunoblot techniques. AB - We report on the development of a novel alternative method for the assessment of floral origin in honey samples based on the study of honey proteins using immunoblot assays. The main goal of our work was to evaluate the use of honey proteins as chemical markers of the floral origin of honey. Considering that honeybee proteins should be common to all types of honey, we decided to verify the usefulness of pollen proteins as floral origin markers in honey. We used polyclonal anti-pollen antibodies raised in rabbits by repeated immunization of Sunflower (Elianthus annuus) and Eucalyptus (Eucalyptus sp.) pollen extracts. The IgG fraction was purified by immunoaffinity. These antibodies were verified with nitrocellulose blotted pollen and unifloral honey protein extracts. The antibodies anti-Sunflower pollen, bound to the 36 and 33 kDa proteins of Sunflower unifloral honey and to honey containing Sunflower pollen; and the antibodies anti-Eucalyptus sp. pollen bound to the 38 kDa proteins of Eucalyptus sp. unifloral honey in immunoblot assays. Satisfactory results were obtained in differentiating between the types of pollen analyzed and between Sunflower honey and Eucalyptus honey with less cross reactivity with other types of honey from different origin and also with good sensitivity in the detection. This immunoblot method opens an interesting field for the development of new antibodies from different plants, which could serve as an alternative or complementary method to the usual melissopalynological analysis to assess honey floral origin. PMID- 11879004 TI - High performance liquid chromatography of phenolic choline ester fragments derived by chemical and enzymatic fragmentation processes: analysis of sinapine in rape seed. AB - High-performance liquid chromatography methods based on reversed-phase chromatography (RPC) and normal phase chromatography (NPC) were introduced for the separation of some representative phenolic acids, choline and betaine, which are the fragments of phenolic choline esters. Sinapine, which is the major phenolic choline ester found in rape seed, was quantitatively hydrolyzed to choline and sinapic acid upon treatment with a solution of sodium hydroxide at room temperature. Choline was further converted to betaine by incubating the base hydrolyzate with choline oxidase. Both sinapic acid and betaine formed the basis for the quantitative determination of sinapine in rape seed by RPC and NPC, respectively. The amounts of sinapine found in rape seed via either of the two fragments (i.e., sinapic acid or betaine) were in very close agreement. PMID- 11879005 TI - Polyethylene glycol, determined by near-infrared reflectance spectroscopy, as a marker of fecal output in goats. AB - We report the application of NIR spectroscopy to determine the fecal concentration of poly(ethylene glycol) (PEG, MW 6000) used as an external marker of fecal output in goats. Calibration was carried out, using the modified partial least-squares method (MPLS), combining all wavelengths in the 1100-2500 nm range, with high linearity (R2 = 0.99). In goats fed at maintenance level, the recovery of PEG in feces was complete, and the estimation of fecal output was accurate, when a moderate dose of PEG was given (20 g/d). A higher dose of PEG (40 g/d) was associated with underestimation of fecal output, probably because PEG interacted with water metabolism. Using PEG and its NIRS-aided analysis to determine fecal output is accurate, simple, and cheap. However, the feasibility of this new method must be verified in goats feeding on tannin-containing diets, and in goats at high feeding level. PMID- 11879006 TI - Polyphenolic antioxidants from the fruits of Chrysophyllum cainito L. (Star Apple). AB - Chrysophyllum cainito L. (Sapotaceae), known commonly as star apple or caimito, is a tropical tree that bears edible fruits. The fruits are grown commercially in certain tropical and subtropical areas, such as southern Florida. In this study, the fresh fruits were extracted with methanol and partitioned with hexane and ethyl acetate sequentially. The ethyl acetate soluble fraction displayed high antioxidant activity in the 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay (IC50 = 22 microg/mL). Activity-guided fractionation of the ethyl acetate soluble fraction was performed to identify the antioxidant constituents. Nine known polyphenolic antioxidants, (+)-catechin (1), (-)-epicatechin (2), (+)-gallocatechin (3), (-) epigallocatechin (4), quercetin (5), quercitrin (6), isoquercitrin (7), myricitrin (8), and gallic acid, have been identified from the fruits. Of these nine antioxidants, 2 is present in the highest concentration in star apple fruits (7.3 mg/kg fresh weight), and 5 showed the highest antioxidant activity (IC50 = 40 microM) in the DPPH assay. PMID- 11879007 TI - Acetylation of 5-amino-1H-[1,2,4]triazole revisited. AB - The products of the acetylation reactions of the common herbicide 5-amino-1H [1,2,4]triazole were investigated using HPLC, GC-MS, 1H NMR, and FTIR spectroscopy. The conventional annular monoacetylation procedures with acetyl chloride are not regioselective and furnish a mixture of isomers. Traditional diacetylation in neat acetic anhydride under reflux produces a mixture of di-, mono-, and triacetylated derivatives. By using equivalent amounts of acetic anhydride in a dimethylformamide solution, a rapid and selective annular monoacetylation of 5-amino-1H-[1,2,4]triazole was achieved. The monoacetylation proceeds via the formation of the intermediate, 1-acetyl-3-amino-1H [1,2,4]triazole, which had not been observed previously and which undergoes transformation into the known 1-acetyl-5-amino-1H-[1,2,4]triazole. Neat acetic anhydride at room temperature affords the diacetylated derivative, 1-acetyl-3 (acetylamino)-1H-[1,2,4]triazole both from 5-amino-1H-[1,2,4]triazole itself and from either 1-acetyl-5-amino-1H-[1,2,4]triazole or 5-(acetylamino)-1H [1,2,4]triazole. The atypical product of the second acetylation, 1-acetyl-5 (acetylamino)-1H-[1,2,4]triazole, has been identified. These results may be useful in the development of effective and selective preparative procedures for the acetylation of 5-amino-1H-[1,2,4]triazole. PMID- 11879008 TI - Antitermitic activity of leaf essential oils and components from Cinnamomum osmophleum. AB - The antitermitic activities of the essential oils from the leaves of two Cinnamomum osmophloeumclones (A and B) and their chemical ingredients against Coptotermes formosanus Shiraki were investigated according to direct contact application. Results from this experiment have demonstrated that the indigenous cinnamon B leaf essential oil has a more effective antitermitic activity than indigenous cinnamon A leaf essential oil. Furthermore, when cinnamaldehyde, eugenol, and alpha-terpineol are extracted from indigenous cinnamon leaf essential oil and used at the strength of 1 mg/g, their antitermitic effectiveness is much higher than that using indigenous cinnamon leaf essential oil. Among the congeners of cinnamaldehyde examined, cinnamaldehyde has exhibited the strongest termiticidal property. PMID- 11879009 TI - Eutypa dieback in grapevines: differential production of acetylenic phenol metabolites by strains of Eutypa lata. AB - The production of acetylenic phenol metabolites in vitro by three strains of the ascomycete Eutypa lata, the causative agent of dying-arm disease in grapevines, has been investigated. Metabolite composition and yields differed significantly between strains and with growth medium but usually reached a maximum after 24-30 days of fungal growth. A general method for the analysis and identification of metabolites by gas chromatography/mass spectrometry of their trimethylsilyl ether derivatives was developed and individual compounds were quantitated by analytical high-performance liquid chromatography (HPLC) and separated by preparative HPLC. The phenolic aldehyde, eutypine (1), reported to be the grape phytotoxin, occurred in only one of the strains examined whereas the primary metabolite was the corresponding alcohol, eutypinol (2), the presumptive detoxification product. A novel metabolite was isolated as a major constituent, together with a minor component, and their structures were established by spectroscopic methods as a methoxyquinol, named eulatinol (4), and a chromene analog (9) of 2, respectively. The evidence suggests that 1 is not solely responsible for phytotoxicity in grapevines but that dying-arm disease may result from a suite of compounds elaborated by the fungus, with the composition dependent on fungal strain and nutritional source. PMID- 11879010 TI - Tyrosinase inhibitors of Pulsatilla cernua root-derived materials. AB - The inhibition of mushroom tyrosinase by Pulsatilla cernua root-derived materials was evaluated. The bioactive components of Pulsatilla cernua root were characterized by spectroscopic analyses as 3,4-dihydroxycinnamic acid and 4 hydroxy-3-methoxycinnamic acid, which exhibited potent antityrosinase activity. The ID50 values of 3,4-dihydroxycinnamic acid and 4-hydroxy-3-methoxycinnamic acid were 0.97 and 0.33 mM, respectively. The compounds isolated from Pulsatilla cernua roots exhibited noncompetitive inhibition against oxidation of L-DOPA by mushroom tyrosinase. This activity was compared with that of three cinnamic acid derivatives and four well-known tyrosinase inhibitors. The ID50 of 4-hydroxy-3 methoxycinnamic acid exhibited superior activity relative to anisaldehyde, anisic acid, benzoic acid, benzaldehyde, cinnamic acid, and cinnamaldehyde; but antityrosinase inhibitors and cinnamic acid derivatives, except for cinnamyl alcohol, were slightly more effective than 3,4-dihydroxycinnamic acid. In the case of benzaldehyde and cinnamaldehyde, the aldehyde group is, apparently, a key group in eliciting potent inhibitory activity, whereas anisaldehyde is more effective than anisic acid. Methoxy substitutions, such as 2-methoxycinnamic acid, 3-methoxycinnamic acid, and 4-methoxycinnamic acid, enhanced inhibition of tyrosinase activity. As a naturally occurring tyrosinase inhibitor, 3,4 dihydroxycinnamic acid and 4-hydroxy-3-methoxycinnamic acid may be useful as new agents to inhibit the oxidation of L-3,4-dihydroxyphenylalanine (L-DOPA) by mushroom tyrosinase. PMID- 11879011 TI - Isoflavone aglycon and glucoconjugate content of high- and low-soy U.K. foods used in nutritional studies. AB - The isoflavone aglycon and glucoconjugate content of commercially prepared and "home-prepared" high- and low-soy foods selected for use in an on-going nutritional study were measured by LC-MS. The daidzin, daidzein, 6"-O malonyldaidzin, 6"-O-acetyldaidzin, genistein, genistin, 6"-O-malonylgenistin, 6" O-acetylgenistin, glycitin, glycitein, 6"-O-malonylglycitin, and 6"-O acetylglycitin content are expressed in terms of individual isoflavones, total isoflavone equivalents, and milligrams of isoflavones per portion served. Soybeans (774 mg x kg(-1) total isoflavones) and soybean-containing foods had the highest isoflavone content of the foods examined. The low-soy foods all contained very low concentrations (<8 mg x kg(-1) total isoflavones) of the isoflavone aglycons and glucoconjugates. High- and low-soy 11 day rotating menus were constructed from the analyzed foods to deliver 100.0 and 0.5 mg of isoflavones per day, respectively. PMID- 11879012 TI - Modulation of cyclizing activity and thermostability of cyclodextrin glucanotransferase and its application as an antistaling enzyme. AB - Cyclodextrin glucanotransferase from Bacillus stearothermophilus ET1 (CGTase ET1) is a potential antistaling enzyme with cyclodextrin (CD)-forming activity. To reduce cyclization activity of CGTase ET1, phenylalanine residues at 191 and 255 were replaced with a glycine (F191G-CGTase ET1) and an isoleucine (F255I-CGTase ET1), respectively. Temperature optima of both mutant enzymes were lower than that of the wild-type. Cyclization activities of both mutants decreased dramatically, but F255I-CGTase ET1 showed a 2-fold higher hydrolytic activity than the wild-type enzyme. CD content of bread loaf treated with F191G-CGTase ET1 was 28.6% of that treated with wild-type, whereas no CD was detected in the loaf treated with F255I-CGTase ET1. Loaves treated with CGTase ET1 or either of the two mutants contained more of the larger maltooligosaccharides such as maltopentaose and maltohexaose than the control and the commercial antistaling enzyme-treated loaves. Retrogradation rates decreased significantly in the loaves treated with either mutant, which indicates the applicability of CGTase ET1 in the bread industry by modulating the cyclizing and hydrolyzing activities of the enzyme. PMID- 11879013 TI - Utilization of enzyme mixtures to retard bread crumb firming. AB - The influence of enzyme mixtures containing amylase and lipase activities on straight dough bread staling was studied. Amylopectin retrogradation, crumb firming, amylose-lipid complexes, and dextrin production were analyzed in bread samples supplemented with two enzyme mixtures. The addition of enzyme mixtures to bread formula causes a beneficial effect on bread keeping properties and the formation of a more thermostable amylose-lipid complex than the one found in control bread. Amylopectin retrogradation was inhibited by the use of the enzyme; the effect was accompanied by reduced crumb-firming rates. The enzymatically generated water-soluble dextrins (maltose and DP3, DP4, DP5, and DP6 dextrins) are the most effective in preserving crumb softness during bread storage. PMID- 11879014 TI - Influence of lysozyme treatments on champagne base wine foaming properties. AB - The objective of this study was to estimate the effect of lysozyme on the foaming characteristics of Champagne base wine. Lysozyme additions were made to the musts and also to the wines before and after bentonite or charcoal treatments, which remove endogenous proteins. Treatments with bentonite diminished foamability and foam stability of wines, whatever the dose (30 or 80 g/hL) and variety [Chardonnay, -28%; Pinot noir, -20% (at 30 g/hL)]. An addition of lysozyme in must raised Pinot noir wine foamability by 21%, whereas the difference is hardly perceptible for Chardonnay wine (+3%). Pinot noir and Chardonnay wines, originating from lysozyme-treated musts, in addition to bentonite treatment on the wine, presented higher foamability than wines treated only with bentonite. Lysozyme was removed (91-100%) by the bentonite treatment. Then, it was not responsible for the increase in foamability but seemed to have a protective effect on the wine proteins. When wines were initially treated with bentonite (150 g/hL) and then enriched with 80 g/hL lysozyme, this enzyme was not able to restore foaming properties. Treatments with charcoal always diminished foamability. The average increase in foamability due to an addition of lysozyme after charcoal treatment (80 g/hL) was 23%. Results showed a real positive effect of lysozyme on foam stability when wines have to be treated with charcoal (+25% and +56% for the Pinot noir wine and the Chardonnay wine, respectively, at 30 g/hL). PMID- 11879015 TI - Chemical characterization of galactomannans and arabinogalactans from two arabica coffee infusions as affected by the degree of roast. AB - Galactomannans and arabinogalactans compose almost exclusively the polysaccharide fraction of roasted coffee infusions. To increase the knowledge about the effect of the degree of roast (DR) in the amount and chemical structure of the galactomannans and arabinogalactans, two arabica coffees of different geographical origins (Costa Rica and Brazil) were roasted for three degrees of roast (DRs 4.7-5.0, 8.7, and 10% of dry weight loss of green coffee beans, on a dry basis). The high molecular weight material was extracted with hot water and dialyzed (molecular weight cutoff >12 kDa), and the material was separated in three cold-water-soluble fractions by graded addition of ethanol. The degree of polymerization and the degree of branching of the galactomannans decreased with the increase of the DR. As the DR increased, less branched arabinogalactans were extracted. The relative amount of terminally linked arabinosyl residues of the arabinogalactans decreased with the increase in DR, and the terminally linked galactosyl residues increased. Also, the size of the arabinosyl side chains of the arabinogalactans decreased with the increase in DR. PMID- 11879016 TI - Microbial inactivation of paprika by a high-temperature short-X time treatment. Influence on color properties. AB - High-temperature short-time (HTST) treatments have been used to destroy the bioburden of paprika. With this in mind, we have designed a device to treat samples of paprika with a gas whose temperature, pressure, and composition can be selected. Temperatures and treatment times ranged from 130 to 170 degrees C and 4 to 6 s, respectively. The survival of the most commonly found microorganisms in paprika and any alteration in extractable and superficial color were examined. Data showed that the optimum HTST conditions were 145 degrees C, 1.5 kg/cm2 of overpressure, 6 s operation time, and a thermal fluid of saturated steam. No microbial growth was detected during storage after thermal treatment. To minimize the color losses, treated (HTST) paprika samples should be kept under refrigeration. PMID- 11879017 TI - Effects of exogenous propylene on softening, glycosidase, and pectinmethylesterase activity during postharvest ripening of apricots. AB - Apricots (Prunus armeniaca L. cv. Boccuccia spinosa) picked at the commercial ripening stage [soluble solids content (SSC) 12.6%] were left to reach full ripening in continuously humidified air at 20 degrees C. Changes in the rate of ethylene production, firmness, soluble solids concentration, and titratable acidity were measured. The alpha-D- and beta-D-glucosidases, alpha-L arabinofuranosidase, alpha-D- and beta-D-galactosidases, beta-D-xylosidase, and alpha-D-mannosidase activities were assayed. To evaluate the influence of ethylene on glycosidase activity, propylene (500 microL x L(-1)) was applied to apricots for 24 and 48 h. In apricots ripened in air, ethylene production increased on the first day and exhibited a typical climacteric pattern. Good edible quality was reached in 5 days when SSC was at least 14% and acidity was between 1.1 and 1.2% (% malic acid). During postharvest ripening, alpha-L arabinofuranosidase activity increased from 1.9 to 11.6 nkat until day 7. alpha-D Galactosidase, alpha-D-mannosidase, and beta-D-galactosidase activity increased continuously but at a lower rate. beta-D-Xylosidase activity also increased, but the level of activity was lower than the other glycosidases assayed. Pectinmethylesterase (PME) decreased during the postharvest ripening, and propylene enhanced this pattern, by stimulating ethylene production. Even the activities of alpha-L-arabinofuranosidase, beta-D-xylosidase, alpha-D mannosidase, and beta-D-galactosidase were greatly stimulated by the propylene treatment, which consequently induced rapid softening of the fruits. PMID- 11879018 TI - Feeding deterrents from Dictamnus dasycarpus Turcz against two stored-product insects. AB - The screening for insecticidal principles from several Chinese medicinal herbs showed that the root bark of Dictamnus dasycarpus possessed significant feeding deterrence against two stored-product insects (Tribolium castaneum and Sitophilus zeamais). From the methanol extract, two feeding deterrents were isolated by bioassay-guided fractionation. The compounds were identified as fraxinellone and dictamnine from their spectroscopic data. Fraxinellone was demonstrated to possess feeding deterrent activity against adults and larvae of T. castaneum as well as S. zeamais adults with EC50 values of 36.4, 29.1, and 71.2 ppm, respectively. Dictamnine was shown to have feeding deterrent activity against adults and larvae of T. castaneum as well as S. zeamais adults with EC50 values of 57.6, 47.9, and 91.7 ppm, respectively. PMID- 11879019 TI - Synthesis, fungicidal activity, and QSAR of pyridazinonethiadiazoles. AB - A series of novel 5-[1-aryl-1,4-dihydro-6-methylpyridazin-4-one-3-yl]-2-arylamino 1,3,4-thiadiazoles, related to the fungicidal activity, were synthesized and tested in vivo against wheat leaf rust, Puccinia recondita. The preliminary bioassay indicated that they exhibited fungicidal activity and the activity was influenced by the nature of the substituents. A quantitative structure-activity relationship study showed that the hydrophobicity (Sigma(pi)) is a major positive parameter in affecting the activity; the electronic parameters (Sigmasigma, SigmaF) are the major negative parameters in affecting the activity. Especially, introducing an ortho substituent with an inductively electron-donating property is favorable to the activity. PMID- 11879020 TI - Zinc transformations in acidic soil and zinc efficiency on maize by adding six organic zinc complexes. AB - Experiments under laboratory and greenhouse conditions were conducted to study the response of maize (Zea mays L.) to Zn fertilizer applications (Zn-phenolate, Zn-EDDHA, Zn-EDTA, Zn-lignosulfonate, Zn-polyflavonoid, and Zn-heptagluconate) in an Aquic Haploxeralf soil. The application of Zn complexes significantly increased Zn uptake by the plant compared with that in the control soil. The highest enhancements were obtained in soil treated with Zn-EDTA, Zn lignosulfonate, and Zn-EDDHA. The highest percentages of Zn taken up by the plants occurred when 20 mg x kg(-1) Zn was applied as Zn-EDTA fertilizer and 10 mg x kg(-1) as Zn-lignosulfonate fertilizer. In the greenhouse experiment, Zn speciation in soil after harvesting showed that almost all Zn was found in the residual fraction followed by metal in the water-soluble plus exchangeable fraction and metal bound to organic matter. The most effective fertilizers maintaining Zn in the most labile fractions were Zn-phenolate, Zn-EDTA, and Zn lignosulfonate. Conversely, in the incubation experiment, only a small percentage of Zn was found in the water-soluble plus exchangeable fraction and no differences in the Zn distribution were observed between the different fertilizer treatments. The micronutrient content in maize was positively correlated with the water-soluble plus exchangeable Zn as well as with the available Zn determined by the diethylenetriaminepentaacetic acid and Mehlich-3 methods, in the greenhouse experiment. Results of this study showed that the incubation experiment in acidic soil is not a suitable tool to establish the different effectiveness of Zn chelates in plants. PMID- 11879021 TI - Residue levels and storage decay control in Cv. Star Ruby grapefruit after dip treatments with azoxystrobin. AB - Cv. Star Ruby grapefruit (Citrus paradisi Macf.) were subjected to a 3-min dip in water at room temperature (20 degrees C) or at 50 degrees C with or without 25, 50, or 100 mg/L azoxystrobin (AZX). Then, the fruits were subjected to cold quarantine at 2 degrees C and 90-95% relative humidity (RH) for 3 weeks and then stored for 5 weeks at 8 degrees C and approximately 85% RH and for another 2 weeks at 20 degrees C and 80% RH to simulate a 2-week marketing period (SMP). No AZX residues were detected in the albedo and pulp following treatments at 20 or 50 degrees C, the total amount of residues being recovered from the flavedo tissue. There was a relationship between the AZX uptake in fruit and the amount of fungicide employed at 20 or 50 degrees C. When AZX was applied to the fruit at 25 mg/L at 20 degrees C, the residue level averaged 0.11 mg/kg (active ingredient, whole fruit basis). This residue concentration increased by 50 and 75% when the application rate increased from 25 to 50 or 100 mg/L, respectively. A similar pattern of accumulation was detected in fruit subjected to treatments at 50 degrees C. However, treatments at 50 degrees C produced residue levels higher than the treatments at 20 degrees C, with increases ranging from 63 to 84%, for the same concentration. Storage conditions did not affect the amount of AZX residues in the fruit. Treatment at 50 or 100 mg/L at 20 degrees C reduced the incidence of moderate to severe chilling injury (CI). Water dips at 50 degrees C reduced the incidence and severity of CI to a very low extent, with no additional advantages when hot water was used in combination with AZX. Treatments with 50 or 100 mg/L of AZX at 20 degrees C produced beneficial effects in decay control similar to those of 25 mg/L AZX at 50 degrees C or hot water alone. Better results were achieved with 50 or 100 AZX at 50 degrees C, providing complete control of decay during cold storage and with negligible decay after SMP. It was concluded that when AZX was applied at 50 degrees C, low doses of fungicide and minimal residue levels in fruit were required to control the postharvest decay of grapefruit. This treatment does not impair fruit quality and offers very interesting prospects for large scale application, due to the reduced potential toxicity of AZX to nontarget organisms and to the environment. PMID- 11879022 TI - Simultaneous determination of furanocoumarins in infusions and decoctions from "Carapia" (dorstenia species) by high-performance liquid chromatography. AB - The use of furanocoumarins, which are photosensitizing compounds, combined with exposure to UV-A radiation is a common treatment for vitiligo, psoriasis, and a number of other skin diseases. Although furanocoumarins plus UV-A treatment is highly effective, several studies have shown that exposure to high doses increases the risk to development of cutaneus carcinoma. Several Dorstenia species are used in folk medicine, mainly against skin diseases, because of the presence of biologically active compounds. We present here analysis of the chemical composition of furanocoumarins from infusion and decoction of "Carapia" (Dorstenia species), which is used in Brazil against several diseases. We have employed high-performance liquid chromatography (HPLC) procedures for the quantitative determination of psoralen, bergapten, and isopimpinellin. The contents of furanocoumarins revealed an insignificant difference between infusion and decoction. Dorstenia tubicina and D. asaroides contained psoralen and bergapten only in the rhizomes, whereas D. vitifolia shows solely isopimpinellin in both rhizomes and aerial parts. PMID- 11879023 TI - Application of artificial aging techniques to samples of rum and comparison with traditionally aged rums by analysis with artificial neural nets. AB - Artificial aging techniques were applied to samples of rum. These samples were then compared, by artificial neural nets, with traditionally aged rums. Analysis was based on the phenolic and furanic composition of each sample. There were found to be few statistical differences between samples, thus confirming the possibility of applying artificial aging techniques to obtain rum with phenolic and furanic characteristics that are similar to those of rum obtained by traditional methods. PMID- 11879024 TI - Original study of the biochemical and oil composition of the Cambodia nut Irvingia malayana. AB - Analysis of the biochemical composition of Irvingia malayana was carried out. This Cambodian nut contains 7.5% water and 70% oil. Most of the fatty acids are saturated and include 42% C12:0 and 41.8% C14:0; the sterol composition is similar to that of other vegetable oils. This oil is less rich in alpha tocopherol than in gamma-tocopherol. Analysis of the solid content of the oil with respect to the temperature by NMR shows a fast fall of solid content around its fusion range at 38-39 degrees C. The main differences in the properties of the indigenous Cambodia nut from other known oleaginous seeds are in its selenium content, fatty acid composition, fusion temperature profile, and content of antioxidants. These important characteristics can soon make possible its application in pharmacology, cosmetics, the margarine industry, etc. PMID- 11879025 TI - Dissipation of the herbicide [14C]dimethenamid under anaerobic conditions in flooded soil microcosms. AB - The objective of this research was to investigate the dissipation of the herbicide dimethenamid under anaerobic redox conditions that may develop in the soil environment. Soil-water biometers were prepared with a saturated soil and made anaerobic by either glucose pretreatment (according to the Environmental Protection Agency registration study for anaerobic fate) or N2 sparging. Treatments included glucose pretreatment, NO3- + SO42- amendment, unamended, and autoclaved. Volatile, aqueous, extractable, and bound (unextractable) 14C residues were quantified and characterized. The redox potential decreased over time, and evidence of denitrifying, iron-reducing, sulfate-reducing, and methanogenic conditions was observed, depending on the amendments. Anaerobic degradation of 14C-dimethenamid occurred in all treatments, and the time observed for 50% disappearance (DT50) was 13-14 days for nonautoclaved treatments. 14C metabolites accumulated to up to 20% of applied 14C. At least two major metabolites were observed in nonautoclaved treatments, whereas only one was observed in autoclaved microcosms. More than 50% of the applied 14C was eventually incorporated into soil-bound residue. PMID- 11879026 TI - Supercritical carbon dioxide extraction and characterization of Laurus nobilis essential oil. AB - Supercritical carbon dioxide extraction allowed essential oil of Laurus nobilis to be obtained. Extraction conditions were as follows: pressure, 90 bar; temperature, 50 degrees C; and carbon dioxide flow, Phi = 1.0 kg/h. Waxes were entrapped in the first separator set at 90 bar and -10 degrees C. The oil was recovered in the second separator working at 15 bar and 10 degrees C. The main components were 1,8-cineole (22.8%), linalool (12.5%), alpha-terpinyl acetate (11.4%), and methyleugenol (8.1%). Comparison with the hydrodistilled oil did not reveal any significant difference. Collection of samples at different extraction times during supercritical extraction allowed the change of the oil composition to be monitored. Lighter compounds such as hydrocarbon and oxygenated monoterpenes were extracted in shorter times than the heavier hydrocarbon and oxygenated sesquiterpenes. PMID- 11879027 TI - Comparison of different methods: static and dynamic headspace and solid-phase microextraction for the measurement of interactions between milk proteins and flavor compounds with an application to emulsions. AB - Interactions between 10 aroma compounds from different chemical classes and 5 mixtures of milk proteins have been studied using static or dynamic headspace gas chromatography and solid-phase microextraction (SPME). Static headspace analysis allows the quantification of the release of only the most abundant compounds. Dynamic headspace analysis does not allow the discrimination of flavor release from the different protein mixtures, probably due to a displacement of headspace equilibrium. By SPME analysis and quantification by GC-MS (SIM mode) all of the volatiles were quantified. This method was optimized to better discriminate aroma release from the different milk protein mixtures and then from oil/water emulsions made with these proteins. The highest difference between the release in different proteins was observed for ethyl hexanoate, which has a great affinity for beta-lactoglobulin. Ethyl hexanoate is thus less released from models and emulsions containing this protein. PMID- 11879028 TI - Formation of methanethiol and dimethyl disulfide in crushed tissues of broccoli florets and their inhibition by freeze-thawing. AB - The formation of methanethiol and dimethyl disulfide in crushed, homogenized, and frozen-thawed tissues of broccoli florets was investigated. These volatile sulfur compounds were produced in crushed florets, but their formation was inhibited in frozen-thawed tissues. Only dimethyl disulfide was formed in homogenized tissues. High pH treatment triggered the release of dimethyl disulfide in frozen-thawed tissues and also enhanced the action of cysteine sulfoxide lyase in all disrupted tissues. Methyl methanethiosulfinate and methyl methanethiosulfonate were not detected in crushed florets; thus, the favored mechanism for the formation of methanethiol and dimethyl disulfide is the chemical disproportionation of methanesulfenic acid. In contrast, the formation of dimethyl disulfide in frozen thawed and homogenized tissues occurs from the chemical disproportionation of methyl methanethiosulfinate that was detected in these tissues. The inhibition of dimethyl disulfide production during freeze-thawing must be caused by a sudden drop in the pH of the tissue, adherence of dimethyl disulfide on the tissue surfaces, and weakening of the cysteine sulfoxide lyase activity under acidic conditions. PMID- 11879029 TI - Aroma extract dilution analysis. Precision and optimal experimental design. AB - The odor thresholds of 12 different compounds have been determined in an AEDA experiment using a panel composed of 8 individuals. Only in one case is the distribution of thresholds among judges significantly different from the log normal. The cause of that departure from normality seems to be a cross adaptation rather than anosmia. The standard deviations (SD) range from 2(0.7) to 2(4.1), with 2(1.8) as average. If the AEDA is carried out at a dilution rate, R, and dilution R(P) (where P = 0, 1, 2.) is the last one at which a compound was detected by a judge, the flavor dilution (FD) factor that should be provided for that judge is R(P+0.5). In the case where several judges carry out the AEDA, the FD for a given compound should be the geometric mean of the FD provided by all the judges. The SD of the distribution of FDs is related to that of the distribution of odor thresholds, but is strongly influenced by the dilution rate, R, being higher with higher R values. The relationship between both SDs can be used to determine the expected precision (measured as the geometric length of the 95% confidence interval) of the result of an AEDA experiment as a function of the dilution rate, the number of judges, and the SD of the distribution of thresholds. Different simulations have shown that in most cases, a dilution rate of 10 is the best option, and that lower dilution rates are advantageous only if the analyzed solution contains compounds with a very narrow distribution of thresholds. PMID- 11879030 TI - Analysis of taste-active compounds in an enzymatic hydrolysate of deamidated wheat gluten. AB - Hydrolyzed plant proteins are widely used as ingredients in culinary products for their glutamate-like ("umami") taste. Three hydrolysates were prepared from wheat gluten using different enzymatic approaches. Comparison of their taste profiles revealed the enzymatic hydrolysate of an acid-deamidated wheat gluten (WGH-3) to be the least bitter of all and to elicit an intense glutamate-like taste. Its umami taste intensity was similar to that of an enzymatic hydrolysate in which glutaminase had been employed to convert free glutamine to glutamic acid and which had a 3-fold higher concentration of free glutamate. Reconstitution studies based on the results of the chemical analysis of WGH-3 and sensory comparison of the model solution and WGH-3 indicated that other components in addition to glutamate and organic acids contribute to its glutamate-like taste. WGH-3 was fractionated by gel permeation chromatography and reversed phase high-performance liquid chromatography, and two fractions with a pronounced glutamate-like taste were obtained. In one of them four pyroglutamyl peptides were tentatively identified: pGlu-Pro-Ser, pGlu-Pro, pGlu-Pro-Glu, and pGlu-Pro-Gln. Apparently, these peptides were formed by cyclization of the N-terminal glutamine residues during the preparation of the hydrolysates. PMID- 11879031 TI - Characterization of the aromatic profile in aqueous essence and fruit juice of yellow passion fruit (Passiflora edulis Sims F. Flavicarpa degner) by GC-MS and GC/O. AB - Chemical characterization by gas chromatography-mass spectometry (GC-MS) of the aromatic profile of yellow passion fruit essence and the juice of the fruit yielded a total of 62 and 34 components, respectively. Four new components have been identified and quantified for the first time in this fruit: 3-methyl-2 butanone; ethyl lactate (quantified only in the fruit juice); diethyl malonate (quantified only in the essence); and 3-penten-2-ol (quantified in both samples). Analysis of these samples by gas chromatography/olfactometry (GC/O) yielded a total of 66 components which appear to contribute to the aroma of passion fruit juice and its aqueous essence. Of these, four could not be quantified by GC-MS: acetic acid, ethyl propionate, ethyl 3-oxobutyrate, and propyl hexanoate. New components were described for the first time as constituents of the aromatic profile in this fruit including acetal, 1,3-dimethyl benzene, and 2-methylbutyl hexanoate. Aroma extract dilution analysis (AEDA) allowed for the detection of the most potent odorants in the commercial essence (2-methylbutyl hexanoate and hexyl hexanoate) and in the fresh juice (1,3-dimethyl benzene and 2-methylbutyl hexanoate). 2-Methylbutyl hexanoate, considered as one of the most potent odorants in this fruit, has been described for the first time as an aromatic constituent of yellow passion fruit. PMID- 11879032 TI - Headspace GC and sensory analysis characterization of the influence of different milk additives on the flavor release of coffee beverages. AB - Previous investigations of coffee flavor have been confined to the analysis of the aroma substances. These investigations showed that about 30 volatile compounds were substantially responsible for the coffee flavor. The aim of this study was to investigate the influence of different milk additives and one coffee whitener on the release of flavor impact compounds from coffee beverages. For the investigation of these effects an external static headspace technique was developed. With this technique the most potent odorants of the coffee beverage were determined. Analyses were performed by gas chromatography/olfactometry, flame ionization detection, and mass spectrometric detection. In addition, sensory studies of the odor profiles were performed. Milk and vegetable products as additives for coffee beverages affected the release of aroma substances in the brew through their lipid, protein, and carbohydrate components. All beverages with an additive showed reduced, but typical, odor profiles for each additive. PMID- 11879033 TI - Selection of yeast starter culture strains for the production of marula fruit wines and distillates. AB - Juice of the Sclerocarya birrea subsp. caffra (marula) fruit was fermented by indigenous microflora and different commercial Saccharomyces cerevisiae yeast strains at different temperatures, namely, 15 and 30 degrees C. Volatile acids, esters, and higher alcohols were quantified in the wine and distillates, and the results were interpreted using a multivariate analysis of variance and an average linkage cluster analysis. Significant differences between 15 and 30 degrees C and also among yeasts with respect to volatile compounds were observed. Yeast strains VIN7 and FC consistently produced wines and final distillates significantly different from the other strains. A panel of tasters and marula and brandy producers was asked to select wines and distillates that had an acceptable and typical marula "nose". They were also asked to detect the differences among wines and distillates fermented with the same yeast strain at different temperatures. PMID- 11879034 TI - Essential oil composition of sachalinmint from Norway detected by solid-phase microextraction and gas chromatography-mass spectrometry analysis. AB - The essential oil of leaves and flowers of sachalinmint [Mentha sachalinensis (Briq.) Kudo] grown in Norway (Trondheim) has been studied by headspace solid phase microextraction (HS-SPME) coupled with gas chromatography-mass spectrometry analysis (GC-MS). The essential oil content increased linearly in acropetal direction from 1.08% (0-20 cm plant height) to 1.75% (60-80 cm; young leaves and flowers). The steam-distilled samples showed a minor complex matrix with a very high menthol and a much lower menthone content (87.89 and 4.05%, respectively). From testing of HS-SPME unequilibrated exposure times ranging from 10 s to 5 min, an extraction time of 30 s was found to be sufficient to detect both low- and high-eluting compounds. Comparison of HS-SPME and steam-distilled samples established that the same tendencies of increasing menthol/menthone content in the basipetal/acropetal direction could be detected by both analysis methods. With regard to the extraction efficiency, HS-SPME gave additional detailed information about less important terpenic compounds. PMID- 11879035 TI - Role of riboflavin in beer flavor instability: determination of levels of riboflavin and its origin in beer by fluorometric apoprotein titration. AB - A method for the quantitative determination of riboflavin levels in beer was developed. The method is based on the quenching of riboflavin fluorescence, which occurs when riboflavin binds to the aporiboflavin-binding protein from egg white. The method does not require any pretreatment of the beer before analysis, other than dilution, and proved to be simple, reliable, and sensitive. The lowest concentration that could be detected was approximately 10 nM riboflavin. The possible interference of flavin mononucleotide (FMN) and flavin adenine dinucleotide (FAD) with the determination of the riboflavin content of beer was excluded, because beer contains only a very small amount of FAD (0.03 microM) and no FMN. The riboflavin levels of the types and brands of beer investigated were in the range of 0.5-1.0 microM. The origin of the riboflavin in beer proved to be the malt. Hop and yeast hardly contributed to the riboflavin content of beer. Besides its use in the determination of riboflavin levels, the aporiboflavin binding protein also provides a way to remove riboflavin from beer, which reduces the light sensitivity and the related lightstruck off-flavor formation in beer. PMID- 11879036 TI - Glycoalkaloid content and chemical composition of potatoes improved with nonconventional breeding approaches. AB - This paper reports the results of chemical analyses performed on two distinct groups of new potato genotypes. The first group contained five clones transformed with the gene ech42 encoding for an endochitinase. The second included 21 interspecific hybrids between the cultivated potato Solanum tuberosum and the wild species S. commersonii, obtained either by somatic fusion or by sexual hybridization. Tubers from transgenic plants were analyzed for several morphological and biochemical parameters to ascertain the substantial equivalence between the transgenic genotypes and the original cultivar Desiree. The interspecific hybrids were analyzed for the same parameters in order to identify genotypes with novel improved chemical characteristics and with low levels of glycoalkaloids deriving from the wild species and potentially hazardous to human health. For transgenic tubers, the results provided evidence that indicates the substantial equivalence between the transgenic genotypes and the cultivated control for the considered traits. The results suggest that chitinase gene insertion did not alter other metabolic pathways of potato tubers and did not cause unintentional pleiotropic effects. As far as interspecific hybrids are concerned, wide variability for all of the parameters analyzed was found. For some useful traits (e.g., soluble solids and proteins, dry matter content) the interspecific hybrids performed better than both the cultivated control and the wild species. In a number of genotypes, glycoalkaloid levels were close to or lower than those of the control varieties, suggesting that selection for low glycoalkaloid content is possible. The results also indicated that glycoalkaloids from S. commersonii may be lost rapidly. Indeed, some hybrids were found to have the same glycoalkaloid profile as S. tuberosum. Finally, the results showed that among the parameters considered, glycoalkaloid content is the most sensitive to variation. Therefore, glycoalkaloid determination should be used for routine control of genotypes produced by interspecific hybridization. PMID- 11879037 TI - Effect of heat-induced aggregation on the IgE binding of patatin (Sol t 1) is dominated by other potato proteins. AB - The interaction of the major potato allergen patatin, Sol t 1, with IgE was investigated on a quantitative level as a function of heat treatment at different temperatures. On the basis of a number of publications, potato is considered to be a heat-labile allergen, but the molecular explanation for this behavior was not given. In this work, heat treatment of patatin in the absence and presence of other potato proteins mimicking the proteinaceous environment of the potato was studied. Using far-UV circular dichrosim spectroscopy, tryptophan fluorescence spectroscopy, and differential scanning calorimetry, the molecular transitions during heating of patatin were investigated. It was found that as long as patatin is not aggregated, denaturation of patatin on a secondary or tertiairy folding level is reversible with only a minor effect on the IgE affinity. Aggregation of patatin results in a nonreversible unfolding and a concomitant important decrease in affinity for IgE (25-fold). Aggregation of patatin in the presence of other potato proteins results in a less condensed aggregate compared to the situation of isolated patatin, resulting in a more pronounced decrease of affinity for IgE (110-fold). It is concluded that the heat lability of patatin-IgE interaction is explained by aggregation of patatin with other potato proteins rather than by denaturation of patatin itself. PMID- 11879038 TI - Heat-induced gel formation by soy proteins at neutral pH. AB - Heat-induced gel formation by soy protein isolate at pH 7 is discussed. Different heating and cooling rates, heating times, and heating temperatures were used to elucidate the various processes that occur and to study the relative role of covalent and noncovalent protein interactions therein. Gel formation was followed by dynamic rheological measurements. Heat denaturation was a prerequisite for gel formation. The gelation temperature (84 degrees C) was just above the onset denaturation temperature of glycinin. The stiffness of the gels, measured as the elastic modulus, G', increased with the proportion of denatured protein. An increase in G' was also observed during prolonged heating at 90 degrees C. This increase is explained by the occurrence of rearrangements in the network structure and probably also by further incorporation of protein in the network. The increase in G' upon cooling was thermoreversible indicating that disulfide bond formation and rearrangements do not occur upon cooling. PMID- 11879039 TI - Hydrogen isotopic profile in the characterization of sugars. Influence of the metabolic pathway. AB - The site-specific natural hydrogen isotope ratios of plant metabolites determined by 2H nuclear magnetic resonance (SNIF-NMR method) can provide powerful criteria for inferring mechanistic and environmental effects on biosynthetic pathways. This work examines the potential of isotopic profiles for the main constituents of carbohydrates, glucose and fructose, to distinguish different photosynthetic pathways. An appropriate analytical strategy, involving three suitable isotopic probes, has been elaborated with a view to measuring simultaneously, in conditions devoid of isotopic perturbations, all (or nearly all) of the carbon bound hydrogen isotope ratios. It is shown that the type of photosynthetic metabolism, either C3 (sugar beet, orange, and grape), C4 (maize and sugar cane), or CAM (pineapple), and the physiological status of the precursor plant exert strong influences on the deuterium distribution in the sugar molecules. Consequently, this isotopic fingerprint may be a rich source of information for the comparison of mechanisms in metabolic pathways. In addition, it can provide complementary criteria to ethanol as a probe for the origin of sugars. PMID- 11879040 TI - Variability in fatty acid and triacylglycerol composition of the oil of coconut (Cocos nucifera L.) hybrids and their parentals. AB - The fatty acid profiles and triacylglycerol (TAG) compositions of oils from the solid endosperm of different Philippine coconut hybrids and their parentals were determined by using gas chromatography (GC) and high-performance liquid chromatography (HPLC). In general, varietal differences in fatty acid composition were observed. Lauric acid (C12) content was significantly higher in the hybrids PCA 15-8 (50.45%) and PCA 15-9 (50.26%) by about 3.16% points as compared to other hybrids, and higher in Tacunan Green Dwarf (50.50%) among the parentals. Among the fatty acids, lauric acid exhibited the least variation. In general, none of the hybrids had higher fatty acid content than their parentals. The HPLC chromatogram of triacylglycerols (TAG) showed 8 major peaks which differ in carbon number (CN) by two: identified as TAG CN 30, 32, 34, 36, 38, 40, 42, and 44. TAGs CN 30 (4.08%) and CN 34 (19.20%) were found to be significantly higher in PCA 15-9 than in the other hybrids. CN 36 was highest (21.94-23.66%) in all hybrids and parentals. The TAG CNs varied significantly among hybrids and parents, i.e., in CN 30, 32, and 34, which are high in medium chain triacylglycerols (MCTs), and in CN 30 (for parentals only), 40, 42, and 44 (the latter two for parentals only), and none in CN 36. MCTs calculated for two hybrids and their parents ranged from 13.81% to 20.55%. PMID- 11879041 TI - Interactions between bovine beta-lactoglobulin and peptides under different physicochemical conditions. AB - The aim of this study was to determine if peptides could interact with beta lactoglobulin (beta-LG) and what the physicochemical conditions promoting their interaction with the protein are. The binding of negatively charged (beta-LG 125 135 and 130-135), positively charged (beta-LG 69-83 and 146-149), and hydrophobic (alphaS1-CN 23-34 and beta-LG 102-105, both bioactive peptides) peptides to bovine beta-LG was determined using an ultrafiltration method under different physicochemical conditions: pH 3.0, 6.8, and 8.0; buffers of 0.05 and 0.1 M; 4, 25, and 40 degrees C; beta-LG/peptide ratios of 1:5 and 1:10. At pH 3.0, none of the peptides interacted with beta-LG at any temperature, buffer molarity, or beta LG/peptide ratio probably due to electrostatic repulsions between the highly protonated species. At pH 6.8 and 8.0, charged peptides beta-LG 130-135, 69-83, and 146-149 bound to beta-LG under some physicochemical conditions, possibly by nonspecific binding. However, both hydrophobic peptides probably bind to the inner cavity (beta-barrel) of beta-LG, provoking the release of materials absorbing at 214 nm. Given the known biological activities of the hydrophobic peptides used in this study (opioid and ACE-inhibitory activities), their binding to beta-LG may be relevant to a better understanding of the physiological function of the protein. PMID- 11879042 TI - Polyphenol/peptide binding and precipitation. AB - Polyphenols are largely responsible for the astringency and "mouthfeel" of tea and wine by their interactions with basic salivary proline-rich proteins. Astringency arises from precipitation of polyphenol/peptide complexes, which is an important protective mechanism in animals that consume polyphenols. This paper presents biophysical studies of the interactions between chemically defined polyphenols and peptides. It is shown that intermolecular binding is dominated by stacking of polyphenolic rings onto planar hydrophobic surfaces and is strengthened by multiple cooperative binding of polyphenolic rings. Affinities weaken at higher temperatures and are unaffected by pH between pH 3.8 and 6.0. Measurements of self-diffusion rates for peptides with increasing concentrations of polyphenol demonstrate that peptides become increasingly coated with polyphenol. When the coating is sufficiently extensive to provide cooperative polyphenol bridges, the peptide dimerizes and precipitates. Light scattering measurements and electron microscopy indicate that the insoluble particles fall into two discrete size classes of ca. 80 and 500 nm diameter. The larger particles are favored at higher temperature and pH, suggesting that the particles are in a colloidal state, with the smaller particles being stabilized by charge repulsion between particles, and that precipitation of the complexes may be a phase separation process. PMID- 11879043 TI - Kinetic study of the acid hydrolysis of various oligofructose samples. AB - The kinetic of acid hydrolysis of five commercially available oligofructose samples used as food ingredients has been investigated as a function of the dry matter concentration, reaction pH, and temperature. The initial fructose release rate is found to be roughly proportional to the inverse of the average polymerization degree in number. A pseudo first order kinetic is found with respect to the fructosyl chain end concentration and to the proton concentration. Arrhenius plot is found to reasonably fit the data in a relatively wide temperature range (7-130 degrees C). The results allow the estimation of the fructose release rate in many foodstuff processing conditions. PMID- 11879044 TI - Diphenol activation of the monophenolase and diphenolase activities of field bean (Dolichos lablab) polyphenol oxidase. AB - This paper reports a study on the hydroxylation of ferulic acid and tyrosine by field bean (Dolichos lablab) polyphenol oxidase, a reaction that does not take place without the addition of catechol. A lag period similar to the characteristic lag of tyrosinase activity was observed, the length of which decreased with increasing catechol concentration and increased with increasing ferulic acid concentration. The activation constant K(a) of catechol for ferulic acid hydroxylation reaction was 5 mM. The kinetic parameters of field bean polyphenol oxidase toward ferulic acid and tyrosine were evaluated in the presence of catechol. 4-Methyl catechol, L-dihydroxyphenylalanine, pyrogallol, and 2,3,4-trihydroxybenzoic acid, substrates with high binding affinity to field bean polyphenol oxidase, could stimulate this hydroxylation reaction. In contrast, diphenols such as protocatechuic acid, gallic acid, chlorogenic acid, and caffeic acid, which were not substrates for the oxidation reaction, were unable to bring about this activation. It is most likely that only o-diphenols that are substrates for the diphenolase serve as cosubstrates by donating electrons at the active site for the monophenolase activity. The reaction mechanism for this activation is consistent with that proposed for tyrosinase (Sanchez-Ferrer, A.; Rodriguez-Lopez, J. N.; Garcia-Canovas, F.; Garcia-Carmona, F. Biochim. Biophys. Acta 1995, 1247, 1-11). The presence of o-diphenols, viz. catechol, L-dihydroxyphenylalanine, and 4-methyl catechol, is also necessary for the oxidation of the diphenols, caffeic acid, and catechin to their quinones by the field bean polyphenol oxidase. This oxidation reaction occurs immediately with no lag period and does not occur without the addition of diphenol. The kinetic parameters for caffeic acid (K(m) = 0.08 mM, V(max) = 32440 u/mg) in the presence of catechol and the activation constant K(a) of catechol (4.6 mM) for this reaction were enumerated. The absence of a lag period for this reaction indicates that the diphenol mechanism of diphenolase activation differs from the way in which the same o-diphenols activate the monophenolase activity. PMID- 11879045 TI - Effect of maturity on chlorophyll, tannin, color, and polyphenol oxidase (PPO) activity of sugarcane juice (Saccharum officinarum Var. Yellow Cane). AB - A study was conducted to determine the effect of sugarcane maturation on the contents of chlorophyll, tannin, and polyphenol oxidase (PPO) activity and on color change of sugarcane juice. The maturation period of the cane studied was between 3 and 10 months after planting. Different parts of the cane, namely, the top, middle, and bottom portions, were analyzed. Results obtained indicated that there were significant (P < 0.01) decreases in total chlorophyll a and b and tannin contents during maturity followed by slower rates of decrease of both parameters at the end of maturity stages. There were no significant differences (P > 0.05) in chlorophyll and tannin contents between the middle and bottom portions. On the other hand, the top portion of the stem had a significantly (P < 0.01) lower concentration of chlorophyll and a significantly (P < 0.01) higher content of tannin. PPO activity of sugarcane juice was determined using chlorogenic acid as a substrate. There was a highly significant difference (P < 0.01) in PPO activity of cane juice during maturity. PPO activity was high at the early development stage, decreased during maturation, and then remained relatively constant at the end of maturity. PPO activity was higher when chlorogenic acid was used as substrate. There were also significant differences (P < 0.01) in juice color (L*, a*, b* values) from different portions at different maturity stages. At the early stages, the color of extracted juice was dark, and then the juice turned to yellowish green during maturity. The decrease in green color or the increase in the yellow color could be associated with the decline in chlorophyll. The overall color change (DeltaE) at maturity indicated that the color of the middle and bottom portions was lower than that of the top portion. PMID- 11879046 TI - Free radical scavenging properties of wheat extracts. AB - Three hard winter wheat varieties (Akron, Trego, and Platte) were examined and compared for their free radical scavenging properties and total phenolic contents (TPC). Free radical scavenging properties of wheat grain extracts were evaluated by spectrophotometric and electron spin resonance (ESR) spectrometry methods against stable 2,2-diphenyl-1-picryhydrazyl radical (DPPH*) and radical cation ABTS*+ (2,2'-azino-di[3-ethylbenzthiazoline sulfonate]). The results showed that the three wheat extracts differed in their capacities to quench or inhibit DPPH* and ABTS*+. Akron showed the greatest activity to quench DPPH radicals, while Platte had the highest capacity against ABTS*+. The ED50 values of wheat extracts against DPPH radicals were 0.60 mg/mL for Akron, 7.1 mg/mL for Trego, and 0.95 mg/mL for Platte under the experimental conditions. The trolox equivalents against ABTS*+ were 1.31 +/- 0.44, 1.08 +/- 0.05, and 1.91 +/- 0.06 micromol/g of grain for Akron, Trego, and Platte wheat, respectively. ESR results confirmed that wheat extracts directly reacted with and quenched free radicals. The TPC were 487.9 +/- 927.8 microg gallic acid equivalents/g of grain. No correlation was observed between TPC and radical scavenging capacities for DPPH* and ABTS*+ (p = 0.15 and p > 0.5, respectively). PMID- 11879047 TI - Release of ferulic acid from oat hulls by Aspergillus ferulic acid esterase and trichoderma xylanase. AB - Oat hulls, an agricultural byproduct, contain a relatively high amount of ferulic acid (FA; 4-hydroxy-3-methoxycinnamic acid), which is believed to be inhibitory to oat hull biodegradability by rumen microorganisms. In this paper, Aspergillus ferulic acid esterase (FAE) was investigated for its ability to release FA from oat hulls. The objectives were to determine the effects of particle size of oat hulls (ground to pass through 1 mm and 250 microm screens and a 100 microm sieve) on release of FA by FAE both in the presence and in the absence of Trichoderma xylanase. The results show that the release of FA by FAE was dependent upon the particle size of oat hulls (< or = 250 microm). In the absence of Trichoderma xylanase, little FA was released by FAE. In the presence of Trichoderma xylanase, there was a significant release of FA by FAE, indicating a synergistic interaction between FAE and Trichoderma xylanase on release of FA from oat hulls. These results indicate that FAE is able to break the ester linkage between FA and the attached sugar, releasing FA from oat hulls. This may leave the remainder of the polysaccharides open for further hydrolytic attack by rumen microorganisms. It is likely that removing FA from oat hulls could improve rumen biodegradability, thus improving the nutritional value of oat hulls. PMID- 11879048 TI - Yeast-induced inhibition of (+)-catechin and (-)-epicatechin degradation in model solutions. AB - (+)-catechin and (-)-epicatechin degradation in water-alcohol solutions containing Fe2+ and tartaric acid was studied in the presence and absence of yeasts. On the basis of the results, yeast partially inhibited the degradation of both flavans, with much slower formation of browning products absorbing at 420 and 520 nm. In comparative terms, yeast was found to be more efficient toward the degradation products of (+)-catechin absorbing at the latter wavelength. Likewise, the presence of yeast decreased the yield of a group of colored compounds eluting at high retention times in HPLC and indicated these as important contributors to color darkening in white wines. This inhibitory effect may in part account for the resistance to browning observed over periods of several years in sherry wines subjected to biological aging under flor yeast. PMID- 11879049 TI - Heat-induced gels of egg white/ovalbumins from five avian species: thermal aggregation, molecular forces involved, and rheological properties. AB - Product processing (heating, pH change, etc.) usually alters protein structure, improves rheological properties, and gives a unique texture to foods. The thermal aggregation and structural properties of ovalbumins from five avian species were studied at different pH values by polyacrylamide gel electrophoresis (PAGE) and determinations of sulfhydryl group content and surface hydrophobicity. The results showed that sulfhydryl group content changed insignificantly in heat denatured ovalbumins other than hen ovalbumin (pH-independent), and surface hydrophobicity markedly increased (pH-dependent) after heating, with a significant difference among species. Furthermore, it was demonstrated that the hydrophobic interaction and sulfhydryl-disulfide interchange reaction were necessary in the aggregation and cross-linking of gel networks. Creep tests were also used to characterize the gel network structures of various egg white/ovalbumins upon heating. The viscoelastic behavior of the ovalbumins of all species was dependent on pH values, and changed significantly with the phylogeny of these species. With increases in pH value (7.0-8.5), the heat-induced gels of ovalbumins gradually changed from turbid to translucent, the instantaneous modulus (E(0)) increased slightly and reached a nearly constant value, and the Newtonian modulus (etaN) increased significantly in each sample. The heated egg white from these five avian species also formed highly viscoelastic gels, with a good correlation of viscoelastic behavior between ovalbumin and egg white in corresponding species. PMID- 11879050 TI - Lower crop load for Cv. Jonagold apples (Malus x domestica Borkh.) increases polyphenol content and fruit quality. AB - The influence of crop load on fruit quality was investigated on 7-year-old slender spindle cv. Jonagold/M.9 apple trees. In mid June five different crop loads per tree were prepared by reducing the fruit number to average 30, 59, 104, 123, and 157 fruits per crown. The fruit from low-cropping trees had more red blush, a higher percentage of soluble solids in fruit flesh, and better flesh firmness in comparison to fruit from high-cropping trees. As the crop load decreased, the concentration of all phenolic compounds in the fruit samples (cortex plus skin) increased; concentrations of the most important individual fruit phenolics were also higher. When crop load fell from 157 to 30 fruits per crown, total polyphenols increased from an average of 1300 to 1680 mg/kg of fruit fresh weight (FW) (+29%), low molecular weight polyphenols increased from 1140 to 1570 mg/kg of FW (+38%), and high molecular weight polyphenols increased from 1740 to 2070 mg/kg of FW (+19%). The average increases in single polyphenols were even greater: chlorogenic acid (+82%); 4'-p-coumaroylquinic acid (+22%); catechin (+178%); and epicatechin (+71%). Ascorbic acid was not significantly dependent on crop load. PMID- 11879051 TI - Reaction of folic acid with reducing sugars and sugar degradation products. AB - The reaction of folic acid with reducing sugars (nonenzymatic glycation) under conditions that can occur during food processing and preparation was studied by high-performance liquid chromatography with diode array detection. N-(p Aminobenzoyl)-L-glutamic acid, a well-established oxidation product, was detected in the reaction mixtures. Furthermore, a new product was isolated and identified as N2-[1-(carboxyethyl)]folic acid (CEF). CEF was the main product that was formed by the nonenzymatic glycation of folic acid. For preparation, N2-[1 (carboxyethyl)]folic acid was obtained in high yields when folic acid and dihydroxyacetone (DHA), a sugar degradation product, were heated at 100 degrees C in phosphate buffer. Mixtures of folic acid and different sugars or DHA were heated under variation of reaction time and temperature, and CEF was quantified. Up to 50% of the vitamin was converted to CEF, with highest yields formed from maltose (49%) and lactose (43%). PMID- 11879052 TI - Deodorization of soybean proteins by enzymatic and physicochemical treatments. AB - To utilize soy protein isolate (SPI) more widely, a convenient and effective method for deodorizing it is required. This paper reports a new deodorizing method using various types of solid adsorbents made of polystyrene, polymethacrylate, and zeolite, as well as charcoal. Treatment of the SPI solution with them decreased the hexanal content in the solution, whereas the content of linoleic acid was not much decreased. A brominated polystyrene adsorbent (SEPABEADS SP207) and a zeolite adsorbent (HSZ-360HUD) removed hexanal most effectively, although 30-40% of the total hexanal remained. A model experiment showed that their hexanal adsorption capacity was much higher than the hexanal content in the SPI solution and that an excess amount of hexanal added to the SPI solution was mostly removed by them. These results suggest that hexanal in the SPI solution can be classified into two types. Hexanal of type I may be free or bound weakly on the surface of proteins and is removable by the adsorbents, whereas hexanal of type II may be bound tightly inside proteins and is unremovable by the adsorbents. Despite the considerable amount of hexanal remaining in the SPI solution even in the most successful cases, the SPI solution was well deodorized as shown by the sensory test. Accordingly, type I hexanal may be closely related to the soybean odor. Removal of hexanal by the adsorbents was not much improved by alpha-chymotryptic digestion of SPI. Type II hexanal might be in similar states even in the chymotryptic digests. PMID- 11879053 TI - 3-deoxypentosulose: an alpha-dicarbonyl compound predominating in nonenzymatic browning of oligosaccharides in aqueous solution. AB - The thermal degradation of D-glucose, maltose, and maltotriose in aqueous solution was investigated under caramelization (no glycine) and Maillard (with glycine) conditions. Degradation of the sugar and alpha-dicarbonyls product was monitored. Under both caramelization and Maillard reaction conditions, 3 deoxypentosulose was the predominating alpha-dicarbonyl compound formed from maltose and maltotriose. In the absence of an amino compound, however, 3 deoxypentosulose is formed in much lower concentration. It was concluded that 3 deoxypentosulose is formed by a pathway specific for oligo- and polysaccharides since this alpha-dicarbonyl is formed from the alpha-1-->4 glucans such as maltose and maltotriose but not from glucose. For its formation, a retro Claisen reaction of an enolization product of 1-amino-1,4-dideoxyhexosulose is proposed as the route to its formation. 1-Amino-1,4-dideoxyhexosulose could be formed by vinylogous alpha-elimination from the 2,3-enediol structure after Amadori rearrangement, favored by planar alignment of the bonds between C1 and C4. Subsequent rearrangement by keto-enoltautomerization leads to a 1-imino-3-keto structure. In this structure, attack of a hydroxyl anion, provided by water at neutral pH, could cause a splitting off of the C1. This reaction gives rise to formic acid or formamide and a pentose derivative, which reacts further to give 3 deoxypentosulose. PMID- 11879054 TI - Peanut roots as a source of resveratrol. AB - A potent antioxidant, resveratrol (3,4',5-trihydroxystilbene), was extracted using 80% methanol from peanut roots (Arachis hypogaea L.), isolated with a solid phase extraction column, purified by a semipreparative HPLC, and identified with 1H NMR and MS. The highest and lowest resveratrol contents in the peanut roots of 2000 fall and 2001 spring crops were 1.330 and 0.130 mg/g and 0.063 and 0.015 mg/g, respectively. When the dehydrated peanut root powders of spring and fall crops were combined and cooked with pork-fat patties (1%, w/w) and the separated oils were stored at 60 degrees C for conjugated diene hydroperoxide (CDHP) determination, CDHP contents of the control oils increased after 3 days of storage, whereas the contents in the peanut root-treated oils of spring and fall crops did not increase after 9 and 15 days of storage, respectively. It is of merit to find that peanut roots, usually left in the field as agricultural waste, contain resveratrol and bear potent antioxidative activity. PMID- 11879055 TI - Processing and cooking effects on lipid content and stability of alpha-linolenic acid in spaghetti containing ground flaxseed. AB - Research was conducted to determine the effect of processing and cooking on the content of hexane-extractable lipid and the stability of alpha-linolenic acid (ALA) in spaghetti fortified with ground flaxseed. Lipid content, ALA, and free fatty acids (FFA) were lower in dried spaghetti samples than in the original semolina-flaxseed mixture. The data indicate that the decline in lipid, ALA, and FFA contents occurred during the extrusion process. In contrast, conjugated diene levels were greater in dried spaghetti than in the corresponding premix. Conjugated diene level was similar for spaghetti samples dried using low- or high temperature drying cycles, and was lower in cooked than in uncooked spaghetti. The low levels of FFA and conjugated diene indicate that ALA remained stable during processing and cooking of spaghetti fortified with ground flaxseed. PMID- 11879056 TI - Direct absorption of acylated anthocyanin in purple-fleshed sweet potato into rats. AB - Absorption of acylated anthocyanins in purple-fleshed sweet potato (Ipomoea batatas cv. Ayamurasaki) in rats was studied to obtain evidence that the acylated anthocyanins themselves could exert a physiological function in vivo. Peonidin 3 caffeoylsophoroside-5-glucoside (Pn 3-Caf*sop-5-glc) in purple-fleshed sweet potato was directly absorbed into rat and present as an intact acylated form in plasma. After oral administration of the purple-fleshed sweet potato anthocyanin (PSA) concentrate containing 38.9 micromol of Pn 3-Caf*sop-5-glc/kg of body weight, Pn 3-Caf*sop-5-glc was detected in the plasma, and the C(max) value and t(max) were estimated as 50.0 +/- 6.8 nmol/Lof plasma and 30 min, respectively. Furthermore, the plasma antioxidant capacity was significantly elevated from 58.0 +/- 12.0 to 89.2 +/- 6.8 micromol of Trolox equivalent/L of plasma 30 min after the administration of the PSA concentrate. PMID- 11879057 TI - Partial purification and kinetic characterization of a carotenoid cleavage enzyme from quince fruit (Cydonia oblonga). AB - For the first time, a cytosolic carotenoid cleavage enzyme isolated from quince (Cydonia oblonga) fruit is described. The enzyme was partially purified by using centrifugation, acetone precipitation, ultrafiltration (300 kD, 50 kD), isoelectric focusing (pH 3-10), and sodium dodecyl sulfate polyacrylamide gel electrophoresis (7.5%). In this way, an enzymatically active protein fraction was obtained that contained three similar proteins, all exhibiting molecular weights in the range of 20 kD. Using beta-carotene as substrate, the enzyme activity was detected spectrophotometrically at a wavelength of 505 nm. The time constant of the reaction was 8.2 min, the Michaelis constant (K(m)) was 11.0 micromol x L( 1), and the maximum velocity (v(max)) was 0.083 micromol x L(-1) x min(-1) x mg(protein)(-1). The optimum temperature was above 50 degrees C. PMID- 11879058 TI - Hydrolytic activity and ultrastructural changes in fruit skins from two prickly pear (Opuntia sp.) varieties during storage. AB - The activity of four cell wall hydrolases, pectinmethylesterase (PME), polygalacturonase (PG), cellulase, and beta-galactosidase (beta-Gal), was measured in fruit skins of two prickly pear varieties, Naranjona and Charola, during storage at 18 degrees C and 85-95% relative humidity (RH). In Naranjona (Opuntia ficus indica), of short postharvest life (ca. 2 weeks), PG, cellulase, and beta-Gal increased their activity more than twice, whereas PME activity tended to increase only slightly during storage. In Charola (Opuntia sp.), of long postharvest life (ca. 2 months), only beta-Gal increased its activity (77%), showing a high PG activity from the beginning of storage. Transmission electron microscopy observations showed middle lamella dissolution at the end of storage for both varieties. Naranjona showed a higher cell wall enzymatic activity than Charola, in agreement with their storability differences. Our results suggest that PG and cellulase in Naranjona and PG and beta-Gal in Charola are the main enzymes responsible for cell wall hydrolytic and ultrastructural changes in skins of stored prickly pears. PMID- 11879059 TI - Detection of a "nonaromatic" NIH shift during in vivo metabolism of the monoterpene carvone in humans. AB - High-resolution gas chromatography in combination with mass spectrometry and high resolution mass spectrometry was used to determine the positions and extent of labeling in the metabolites of carvone, namely in alpha,4-dimethyl-5-oxo-3 cyclohexene-1-acetic acid (dihydrocarvonic acid), alpha-methylene-4-methyl-5-oxo 3-cyclohexene-1-acetic acid (carvonic acid), and 5-(1,2-dihydroxy-1-methylethyl) 2-methyl-2-cyclohexen-1-one (uroterpenolone), after human ingestion of 9,9 dideutero- and 9-(13)C-carvone. Carvonic acid was formed by oxidation at the methyl carbon of the isopropenyl group of carvone, whereas dihydrocarvonic acid was formed by oxidation at the methylene position, most probably via carvone epoxide. A "nonaromatic" NIH shift must occur during the subsequent reactions yielding dihydrocarvonic acid. Additionally, dehydrogenation of dihydrocarvonic acid and hydrogenation of carvonic acid were observed, resulting in minor amounts of both acids owning a carboxy group of opposite origin. Uroterpenolone was found to be exclusively formed by oxidation at the methylene carbon of the isopropenyl group of carvone, and thus, most probably by hydrolysis of carvone epoxide. PMID- 11879060 TI - Milk-arterial plasma transfer of PCDDs and PCDFs in pigs. AB - Polychlorodibenzo-para-dioxins (PCDDs) and Polychlorodibenzofurans (PCDFs) are ubiquitous in the environment. They are mainly formed as unwanted byproducts during various chemical, industrial, and combustion processes. Thus, these pollutants can be found in the food chain. The aim of this experiment was to study the transfer of PCDD/Fs from spiked milk to arterial blood in pigs, which are considered as valid models for humans. Pigs were fed with 900 mL of milk spiked with a mixture of 17 dioxins. The levels of PCDD/Fs in the serum extracts were determined using HRGC/HRMS prior to consumption of the milk, and at 3, 5, and 7 h after milk ingestion. Concentrations of PCDD/Fs in arterial plasma increased from 3 h to 5 h and decreased at 7 h. At time point 5 h, concentrations were found between 500 and 10000 pg x g(-1) fat. The transfer ratio <> was usually found between 0.7 and 3%. Related to the different milk concentrations, results of this study indicate a similar behavior of the studied molecules. PMID- 11879061 TI - Stability of the flavan-3-ols epicatechin and catechin and related dimeric procyanidins derived from cocoa. AB - Cocoa flavanols and procyanidins possess wide-ranging biological activities. The present study investigated the stability of the cocoa monomers, (-)-epicatechin and (+)-catechin, and the dimers, epicatechin-(4beta-8)-epicatechin (Dimer B2) and epicatechin-(4beta- 6)-epicatechin (Dimer B5), in simulated gastric and intestinal juice and at different pH values. The dimers were less stable than the monomers at both acidic and alkaline pH. Incubation of Dimer B2 and Dimer B5 in simulated gastric juice (pH 1.8) or acidic pH resulted in degradation to epicatechin and isomerization to Dimer B5 and Dimer B2, respectively. When incubated in simulated intestinal juice or at alkaline pH, all four compounds degraded almost completely within several hours. These results suggest that the amount, and type, of flavanols and procyanidins in the gastrointestinal tract following the consumption of cocoa can be influenced by the stability of these compounds in both acidic and alkaline environments. PMID- 11879062 TI - Combination of lipids and emulsifiers enhances the absorption of orally administered quercetin in rats. AB - The effects of lipids, emulsifiers, and ethanol on the absorption of orally administered quercetin in rats were investigated for its efficient intestinal absorption. Rats were administered 150 micromol/kg quercetin in water supplemented with lipids and/or emulsifiers, or ethanol, and blood was collected from the tail for 6 h after administration. Co-administration of lipids such as lecithin and soybean oil or emulsifiers including sucrose fatty acid ester, polyglycerol fatty acid ester, and sodium taurocholate had no statistically significant effects on quercetin absorption, although these constituents rather increased the accumulation of conjugated forms of quercetin and those of isorhamnetin in rat plasma. However, the combination of lipids and emulsifiers enhanced the absorption of quercetin significantly. Thirty and fifty percent (v/v) of the ethanol in the vehicle raised the efficiency of quercetin absorption in a concentration-dependent manner. Quercetin absorption-enhancing effects of these constituents seemed to be affected by quercetin's solubility in respective vehicles used for the administration. Ethanol is not helpful for the effective absorption of quercetin, as a high concentration is required. In conclusion, a combination of lipids and emulsifiers is necessary for enhancing quercetin absorption. PMID- 11879063 TI - Isolation and characterization of undenatured chlorogenic acid free sunflower (Helianthus annuus) proteins. AB - A method for obtaining sunflower protein (SFP) isolate, nondenatured and free of chlorogenic acid (CGA), has been developed. During the isolating procedure, the extent of CGA removal and protein denaturation was monitored. The defatted flour contained 2.5% CGA as the main phenolic compound. Phenolic compounds were removed by aqueous methanol (80%) extraction, before protein extraction at alkaline pH and diafiltration. Differential scanning calorimetry and solubility tests indicated that no denaturation of the proteins had occurred. The resulting protein products were biochemically characterized, and the presence of protein CGA complexes was investigated. SFPs of the studied variety were found to be composed of two main protein fractions: 2S albumins and 11S globulins. In contrast to what has been previously reported, CGA was found to elute as free CGA, not covalently associated to any protein fraction. PMID- 11879064 TI - Liquid chromatography-mass spectrometry (LC-MS) investigation of the thiobarbituric acid reactive substances (TBARS) reaction. AB - The thiobarbituric acid reactive substances (TBARS) assay is a commonly used method for the detection of lipid peroxidation. Malondialdehyde is formed as a result of lipid peroxidation and reacts with thiobarbituric acid to form a pink pigment that has an absorption maximum at 532 nm. Other compounds also react with thiobarbituric acid to form colored species that can interfere with this assay, but little is known about these interfering species. This is the first investigation using LC-MS and MS-MS to study the structures of the pink adduct as well as a common unstable yellow interference compound, which absorbs at 455 nm. Also, the presence of barbituric acid impurities in the thiobarbituric acid reagent was found to produce 1:1:1 thiobarbituric acid/malondialdehyde/barbituric acid and 2:1 barbituric acid/malondialdehyde adducts that absorbed at 513 and 490 nm, respectively, indicating that thiobarbituric acid should be purified before use. PMID- 11879065 TI - Quercetin derivatives are deconjugated and converted to hydroxyphenylacetic acids but not methylated by human fecal flora in vitro. AB - By using a batch in vitro anaerobic fecal fermentation model, we have shown that the fecal microflora can rapidly deconjugate rutin, isoquercitrin, and a mixture of quercetin glucuronides. High levels of beta,D-glucosidase, alpha,L rhamnosidase, and beta,D-glucuronidase were present. Rutin underwent deglycosylation, ring fission, and dehydroxylation. The main metabolite, 3,4 dihydroxyphenylacetic acid, appeared rapidly (2 h) and was dehydroxylated to 3 hydroxyphenylacetic acid within 8 h. The pattern of in vitro fermentation of rutin was not changed by changing the pH (6.0 or 6.9), fermentation scale (10 or 1000 mL), or donors of the inoculum. Hydroxyphenylacetic acids were not methylated by colon flora in vitro. The colonic microflora has enormous potential to transform flavonoids into lower molecular weight phenolics, and these might have protective biological activities in the colon. The site of absorption of flavonoids and the form in which they are absorbed are critical for determining their metabolic pathway and consequent biological activities in vivo. PMID- 11879066 TI - Effect of dietary protein quantity and quality on the brain protein synthesis rate in ovariectomized female rats. AB - The purpose of this study was to determine whether the quantity and quality of dietary protein affected the rate of brain protein synthesis in ovariectomized female rats. Two experiments were conducted on the ovariectomized female rats (12 weeks old) given diets containing 20%, 5%, or 0% casein (experiment 1) and 20% casein, 20% soy protein, 20% gluten, or 20% gelatin (experiment 2) for 10 d, respectively. The fractional rates of protein synthesis in the brain declined with a decrease of the quantity and quality of dietary protein. In the brain, the RNA activity [g of protein synthesized/((g of RNA) d)] was significantly correlated with the fractional rate of protein synthesis. The RNA concentration (mg of RNA/g of protein) was not related to the fractional rate of protein synthesis in any organ. The results suggest that the rate of protein synthesis in the brain declines with the decrease of the quantity and quality of dietary protein in ovariectomized female rats, and that RNA activity is at least partly related to the fractional rate of brain protein synthesis. PMID- 11879067 TI - Occurrence of zearalenone-4-beta-D-glucopyranoside in wheat. AB - An LC-MS method was developed for the analysis of zearalenone-4-beta-D glucopyranoside and zearalenone in wheat (Triticum aestivum). The limit of determination for zearalenone-4-beta-D-glucopyranoside and zearalenone was 10 microg/kg. The recovery rates were calculated to be 69% and 89% at a concentration of 100 microg/kg for zearalenone-4-beta-D-glucopyranoside and zearalenone, respectively. Twenty-four Bavarian wheat samples from a 1999 harvest were analyzed. Zearalenone was present in 22 of 24 field samples, the levels ranged from 11 to 860 microg/kg. Zearalenone-4-beta-D-glucopyranoside was found in 10 of the zearalenone positive samples (42%) at levels ranging from 17 to 104 microg/kg. The amounts of zearalenone-4-beta-D-glucopyranoside were correlated to those of zearalenone (r2 = 0.86, b = 0.10). After gastrointestinal hydrolyzation, zearalenone-4-beta-glucopyranoside might be implicated in the development of a zearalenone-syndrome. Therefore, more attention should be focused on conjugated mycotoxins in food and feed. PMID- 11879068 TI - Magnetic resonance imaging of the levator veli palatini muscle during speech. AB - OBJECTIVE: To obtain detailed anatomic information on the levator veli palatini (LVP) muscle from magnetic resonance imaging (MRI). Quantitative measures of the configuration of the LVP muscle at rest and during speech activities were obtained. DESIGN: Prospective study using MRI of adult subjects with normal velopharyngeal mechanisms to determine anatomic and physiologic parameters of the levator muscle. The levator veli palatini muscle was imaged at rest and during speech activities consisting of nasal and non-nasal sounds mixed with vowels, consonants, or both (e.g., /ansa, asna, amfa, afma/). PARTICIPANTS: Ten normal healthy adults (five men, five women) between 21 and 53 years of age and free of oropharyngeal abnormalities. MAIN OUTCOME MEASURES: Two-dimensional spin echo static images and dynamic fast gradient echo images of the levator muscle in both the sagittal and oblique/coronal planes. RESULTS: On average across female (F) and male (M) subjects: distance between LVP muscle origin points, 52.6 mm (F), 54.6 mm (M); angle of levator muscle origin at rest, 64.5 degrees (F), 60.4 degrees (M); length of the levator muscle at rest, 44.1 mm (F), 46.4 mm (M); width of levator muscle at lateral margin of velum, 5.5 mm (F), 6.6 mm (M). Both the levator muscle angle of origin and length became progressively smaller from rest, nasal consonants, low vowels, high vowels, and fricatives for both female and male subjects. Across all subjects, there was a 19% reduction in length of the LVP muscle from rest position to fricative production. CONCLUSIONS: MRI is an effective method of imaging and measuring the LVP muscle and related structures in living subjects. Understanding the normal tissue distribution and quantification of the LVP muscle provides important information for development of a functional biomechanical model of the velopharynx and for improved surgical treatment. PMID- 11879069 TI - Airway obstruction following palatoplasty: analysis of 247 consecutive operations. AB - OBJECTIVE AND METHODS: Between February 1987 and September 1997, 247 patients underwent primary repair of a cleft of the secondary palate by one surgeon, using the double-opposing Z-plasty (Furlow) technique. This retrospective study reviews perioperative and postoperative airway compromise among these patients. The purposes of this study were to identify factors associated with airway obstruction following palatoplasty and to analyze the management of those patients. Although infants experiencing airway problems following Wardill-Kilner and Von Langenbeck palatoplasty have been described, airway complications in a group of Furlow repair patients has not been previously reported. RESULTS: Fourteen patients (5.7%) had airway problems. The average age of these patients was 18 months, which was not significantly different from those without airway problems. Airway obstruction occurred as late as 48 hours after the completion of surgery. Twelve of the 14 patients had severe airway compromise requiring continued postoperative intubation, reintubation, or tracheostomy (one). There were no deaths. Thirteen of the 14 patients with postoperative airway problems (93%) had other congenital anomalies in addition to clefting, a named congenital disorder, or both. Seven of those 13 had Pierre Robin sequence. In contrast, only 40 of the 233 patients without airway problems (17%) had additional congenital anomalies or named disorders. Presence of other congenital anomalies was associated with a significantly increased risk of airway obstruction (p =.005). CONCLUSION: Patients with cleft palate with the Pierre Robin sequence or other additional congenital anomalies had an increased risk of airway problems following palatoplasty. Awareness of this risk permits identifying those patients prior to surgery so that they can be monitored and managed appropriately, minimizing the likelihood of major complications or death. PMID- 11879071 TI - The impact of early palatal obturation on consonant development in babies with unrepaired cleft palate. AB - OBJECTIVE: The purpose of this investigation was to determine whether palatal obturators enhance consonant development during babbling for babies with unrepaired cleft palate. PARTICIPANTS: Fourteen babies with cleft palate who had worn anterior palatal obturators prior to palatal surgery were matched to 14 unobturated babies according to cleft type, sex, and age at time of presurgical evaluation. MAIN OUTCOME MEASURES: Spontaneous vocalizations of the obturated and unobturated groups were compared to determine whether differences were evident in size of consonant inventory as well as place and manner of consonant production. RESULTS: Paired t tests revealed no significant differences between the groups in size of consonant inventory or place and manner of consonant production. There was a trend for babies in the obturated group to produce more glottal consonants. CONCLUSIONS: In general, the findings of this study suggested that palatal obturators do not appear to facilitate production of anterior palatal consonants during babbling. PMID- 11879070 TI - Nonsyndromic cleft lip with or without cleft palate in China: assessment of candidate regions. AB - OBJECTIVE: Although Asians have the highest birth prevalence of oral-facial clefts, the majority of gene mapping studies of cleft lip with or without cleft palate (CL/P) have been in European or American Caucasians. Therefore, the objective of this study of Chinese families was to evaluate linkage and association between CL/P and 10 genetic markers in five chromosomal regions that have shown positive results in Caucasians. SETTING: Families were ascertained through nonsyndromic CL/P surgical probands from hospitals throughout Shanghai, China. PARTICIPANTS: Study participants included 671 individuals from 60 families with two or more members affected with oral-facial clefts. Of the 671 total individuals, 145 were affected. RESULTS: Ten markers from chromosomes 2, 4, 6, 17, and 19 were assessed (TGFA, MSX1, D4S194, D4S175, F13A1, GATA185H, D17S250, D17S579, D19S49, APOC2). LOD scores were calculated between each of the 10 markers and CL/P as well as model-free statistics of linkage (SimIBD) and association (TDT). None of the markers showed significantly positive LOD scores with CL/P. A significantly positive result (p =.01) was seen using SimIBD for APOC2 on chromosome 19, and a positive TDT result (p =.004) was obtained for D19S49, near APOC2. CONCLUSIONS: This is the first gene mapping study of CL/P in China. These results indicate that most of the genetic regions with positive results in Caucasian families may not be involved in CL/P found in China, although there is some positive evidence for the candidate region on chromosome 19. PMID- 11879072 TI - Beyond easy answers: prenatal diagnosis and counseling during pregnancy. AB - OBJECTIVE: The advancing sophistication and availability of prenatal diagnostic technologies, such as transvaginal ultrasound, chorionic villus sampling, amniocentesis, and alpha feto-protein testing, have increased the medical capacity to detect genetic and congenital conditions during pregnancy. This paper raises many social and ethical questions about how families, craniofacial teams, and society respond when a prenatal diagnosis is made and considers the ethical and social issues around counseling, managing information, and making decisions. DESIGN: Ethical and sociological analysis. SETTING: Implications examined on the societal, health professional, and family level. RESULTS: Families and health professionals often manage ambiguity, uncertainty, and complex decision making in facing a prenatal diagnosis. Embedded in parental and clinical decisions are values about children with birth defects. Families are making decisions about whether to bear or abort an affected fetus on the basis of their perceptions of the impairment and on their expectation of the burden involved for the family and the child. On a broader, societal level, pressures to conform and minimize human differences are apparent in biomedical interventions, the Human Genome Project, advertising and media images, and social pressures to normalize disabilities. How society deals with prenatal diagnosis will impact upon social values; moral, legal, and ethical perspectives; and on health policy. CONCLUSION: Prenatal diagnostic technologies raise complex ethical, family, policy, and legal issues that have broad implications for the lives of children born with special health care needs, including children with cleft lip and palate. PMID- 11879073 TI - Prenatal diagnosis of cleft lip and palate: detection rates, accuracy of ultrasonography, associated anomalies, and strategies for counseling. AB - OBJECTIVE: As ultrasound becomes more widely utilized in pregnancy and imaging technology improves, cleft lip and palate will become more commonly identified in prenatal life. In efforts to meet the needs for information regarding cause and management, pregnant women and their partners are increasingly referred to cleft and craniofacial treatment programs. This group of patients provides unique challenges to professionals unfamiliar with the issues inherent to this population. Information regarding the extent of the defect and the absence of associated abnormalities is usually incomplete. Treatment teams may be uncomfortable with the possibility that couples may choose not to continue a pregnancy on the basis of what they hear. Currently between 14% and 25% of cleft lip, with or without cleft palate, is detected antenatally. About 12% of presumably isolated clefts will be one feature in a broader pattern of malformation. This article reviews the current status of ultrasound in the detection of clefts during pregnancy and outlines a strategy for counseling based on the author's experience in both a prenatal diagnosis program and a cleft craniofacial treatment team. PMID- 11879074 TI - Prenatal diagnosis: when the clinician disagrees with the patient's decision. AB - OBJECTIVE: Recent reports of prenatal diagnosis of cleft lip and cleft palate resulting in pregnancy termination have raised questions about the appropriateness of such decisions. This review attempts to clarify the importance of nondirective counseling and define the role of the clinician, especially in the face of difficult and often controversial decisions. DESIGN: Genetic counseling literature was reviewed to clarify the role of genetic counseling in prenatal diagnosis within the framework of different counseling approaches. CONCLUSIONS: The ultimate decision regarding pregnancy outcome and management, including termination, belongs to the pregnant couple. Clinicians will often have their own thoughts about what decisions they would make when faced with certain clinical situations. However, the responsibility of clinicians is to help the couple make their own decisions based upon their personal religious, moral, and cultural values. PMID- 11879075 TI - Beyond easy answers: the plastic surgeon and prenatal diagnosis. AB - OBJECTIVE: To explore how plastic surgeons handle the issue of prenatal diagnosis of cleft lip and palate. DESIGN: Mail survey. PARTICIPANTS: The plastic surgeon listed as team contact in the team directory of the American Cleft Palate Craniofacial Association was identified. In cases in which a plastic surgeon was not listed as a team contact, the first plastic surgeon listed as a team member was identified. A total of 211 surgeons were contacted. INTERVENTION: Each surgeon was mailed a survey asking for his/her experience in prenatal diagnosis for cleft conditions. Ethical issues concerning the surgeon's personal beliefs about abortion and the potential influence of the surgeon's presentation of information were explored. RESULTS: A 50% response rate was obtained. Eighty-five percent of respondents were experienced in prenatal counseling; 92% offer consults on a routine basis. Ninety-six percent indicated they did so because they felt it was helpful to families. Treatment (98%), appearance of clefts (96%), cleft etiology (94%), and associated problems (40%) were discussed most commonly. Termination was rarely discussed (5%). Of those who had not met with a family, 82% said they had never been asked to do so. Most respondents felt families sought consultation for information and rarely sought consultation to decide on abortion. The majority felt their personal beliefs about abortion did not influence their willingness to meet with parents. The majority of surgeons in both groups felt it was ethical to give an opinion on pregnancy termination if asked (55% of experienced surgeons, 44% of inexperienced surgeons); however, a significantly larger number of the surgeons who had not met with a family were unsure of their opinion. CONCLUSIONS: Prenatal counseling of parents of a fetus with cleft lip/palate is common among plastic surgeons. There is uniformity in the information discussed. There is no agreement among surgeons about the ethical issues of pregnancy termination and the surgeon's role in that discussion. PMID- 11879076 TI - Dealing with the prenatal diagnosis of clefting: a parent's perspective. AB - OBJECTIVES: Prenatal testing and information is available on an increasing basis for a variety of reasons. There are conflicting standards of care in the delivery of this type of evaluation and even more confusion around access to such services. As physicians and patients evaluate which tests will generate useful information with acceptable levels of risk, the rules of the game constantly shift. Presenting the situation from the viewpoints of both a professional and a parent, issues revolving around uncertainty and reassurance are evaluated. A description of the personal experience of the impact of the discovery of a cleft prenatally is provided. CONCLUSIONS: Finally, efforts are made to raise questions about our motivations behind conducting prenatal testing and also developing some suggestions about how to support families better in our regular clinical work. A challenge is made to providers and patients to do more work prior to examination to explore how the unexpected will be handled. The dilemma about how to facilitate informed consent through full disclosure without overwhelming the patient is also discussed. PMID- 11879077 TI - Use of US birth certificate data to estimate the risk of maternal cigarette smoking for oral clefting. AB - OBJECTIVE: The purpose of this study was to evaluate the relationship between maternal cigarette smoking and the risk of having an offspring with an oral cleft. DESIGN: This was a large population-based, matched case-control study derived from the United States Natality database for 1997. SUBJECTS: The sample consisted of 2029 cases with non-syndromic oral clefts and 4050 non-malformed controls. Controls were matched to cases on mother's and father's race and child's sex, county of birth, and month of birth. This sample was selected from a total of 3,093,821 births in the United States, which represents 80% of all births in this country during 1997. RESULTS: The association between maternal cigarette smoking and oral clefts in the offspring was close to the null (odds ratio 1.16; 95% confidence interval (CI) 1.01 to 1.33; one-sided Fisher exact test p =.0207). The comparison and pooling of results to those of a similar study that used the U.S. Natality database for 1996 resulted in a common Mantel Haenszel odds ratio of 1.33 (95% CI 1.21 to 1.46). The dose-response analysis was slightly significant for all levels of maternal smoking. CONCLUSION: This large study confirms that smoking during pregnancy is only a minor risk factor for oral clefting in the offspring. PMID- 11879078 TI - Dental development of permanent lateral incisor in complete unilateral cleft lip and palate. AB - OBJECTIVE: The objective of this retrospective study was to verify whether the root development of the permanent lateral incisor on the cleft side was delayed, compared with its contralateral tooth in subjects with complete unilateral cleft lip and palate. SETTING: Craniofacial Anomalies Rehabilitation Hospital (HRAC) University of Sao Paulo-Bauru, Brazil. PATIENTS: A sample of 98 panoramic radiographs of HRAC patients with complete unilateral cleft lip and palate was selected, regardless of sex and race. MAIN OUTCOME MEASURE: Panoramic radiographs were analyzed and the root development stage of the cleft-side permanent lateral incisor was compared with its contralateral tooth. RESULTS: Overall, the permanent cleft-side lateral incisor was significantly delayed in root development in relation to the non-left side contralateral tooth (p <.05). However, no significant differences were observed between boys and girls. CONCLUSIONS: Root development of the permanent cleft-side lateral incisor was delayed, compared with its contralateral tooth. The cleft itself may play an important role in this delay because the results from boys and girls were very similar. PMID- 11879079 TI - Effects of calcium phosphate ceramic bone graft materials on permanent teeth eruption in beagles. AB - OBJECTIVE: The aim of this study was to evaluate the use of calcium phosphate ceramic (CPC) materials as a potential alternative to autogenous secondary alveolar bone grafting in cleft lip and palate patients who are in mixed dentition. SAMPLE: Four 12-week-old beagles and one 15-week-old beagle were used as subjects. INTERVENTIONS: In each experimental beagle, the third and fourth deciduous premolars were extracted. The sockets were filled with four different CPC materials and sutured. MAIN OUTCOME MEASURES The beagles were fed a soft diet for the following 8 weeks and then sacrificed for clinical, radiological, histological, transmission electron microscope (TEM), and infrared (IR) absorption analysis. RESULTS: All four experimental graft materials allowed normal development and eruption of permanent premolars. In histological sections, small particles of biphasic calcium phosphate (BCP) and carbonate apatite (CO(3) AP) were resorbed, and large particulate forms served as bone frames in cortical bones. Polymer coated with carbonate apatite (Poly/CO(3)-AP) did not cause inflammation but was pushed away to the soft tissue by erupting teeth. Alginate coated with carbonate apatite (alginate/CO(3)-AP) caused a severe inflammatory reaction to the point of destroying a part of the dental follicle and cortical bone. In TEM, resorption activity by phagocytic cells was observed only in CO(3) AP. Direct bonding of CO(3)-AP to the bone was observed as the electron-dense interface between bone and CO(3)-AP. CONCLUSION: BCP and CO(3)-AP proved to be suitable as alveolar bone graft materials in areas where tooth eruption occurs. Of the four materials tested, CO(3)-AP produced the best results. PMID- 11879080 TI - Craniofacial anthropometry in a Turkish population. AB - OBJECTIVE: To present norms and demonstrate the anthropometric variations in fronto-occipital circumference, inner and outer canthal distances, near and distant [far] anatomical interpupillary distance, canthal index, and circumference-interorbital index across age and sex in urban Turkish subjects. PARTICIPANTS: Three thousand four hundred forty-eight subjects (1852 male, 1596 female) aged 7 to 40 years were included in this study. METHODS: Three age groups were studied: children aged 7 to 15 years, young adults aged 16 to 25 years, and adults aged 26 to 40 years. Mean values for each measured parameter were determined at each age between 7 and 25 years. Subjects were also divided into nine age subgroups to observe the change of each parameter with advancing age. RESULTS: The fronto-occipital circumference and outer canthal distance of males was significantly (p <.001) wider than females in all age groups. The near and distant interpupillary distances of male subjects were, on average, wider than the female subjects with greater differences with advancing age. Across all subjects aged 7 to 40 years, the mean of all measured parameters and calculated indexes of men and boys was significantly different from girls and women (p <.001). The mean for interpupillary distances in our study in both sexes were found to be similar to Arabian, Hong Kong, and British children; larger than those of Chinese, Black, Indian, and Caucasians; and smaller than those of Mexican children and a mixed European population. CONCLUSION: This study clearly shows the anthropometric variation for fronto-occipital circumference, inner canthal distance, outer canthal distance, near and distant interpupillary distance, canthal index, and circumference-interorbital index with age. These developmental data and the normal values of these measurements in healthy subjects are useful for dysmorphologists in the early identification of some craniofacial syndromes, hyper- and hypotelorism, and congenital or posttraumatic telecanthus and of planning surgical intervention. We suggest that the comparison of craniofacial dimensions of a patient must be performed with normal standards specific for age as well as sex and race. PMID- 11879081 TI - A cephalometric study of facial growth in van der Woude syndrome. AB - OBJECTIVE: To test the hypothesis that maxillary growth and lower lip form in patients with van der Woude syndrome (VDW) is different from patients with non syndromic cleft lip and palate. DESIGN AND SETTING: Retrospective, case-control study at a tertiary cleft center, Chang Gung Memorial Hospital, Taipei, Taiwan. PATIENTS AND PARTICIPANTS: Records of 53 patients with VDW, who presented for treatment during the years 1968 through 1998 were obtained. Twenty-three of 53 patients had received at least one lateral cephalogram during the course of their treatment. Of these 23, in 17 it was possible to find non-syndromic case controls with identical cleft type, sex, and method of cleft palate repair, with year of birth matching within 1 year of the corresponding VDW patient. For these 17 pairs of VDW and non-syndromic cleft controls, cephalogram acquisition dates were checked to see how well the corresponding pairs matched. A total of 43 pairs of cephalograms were deemed to be acceptably matched because the ages at acquisition differed by less than 15% of the VDW patient's age. MAIN OUTCOME MEASURES: Thirteen measurements were derived from the 11 standard lateral cephalometric landmarks recorded on each cephalogram. The data were classified into five groups according to age at time of cephalogram, and sets of paired non-syndromic cleft and VDW measurements were tested for differences using a Wilcoxon signed rank sum test in two ways, first including all cleft types and then including only those patients with complete bilateral cleft lip and palate. A longitudinal growth analysis considering the movement of the skeletal A and B points was performed on the patients with complete bilateral cleft lip and palate. RESULTS: For the osseous measurements, anteroposterior maxillary length as described by the anterior nasal spine (ANS)-posterior maxillary point distance was statistically significantly shorter in the VDW patients of age 13 years and older, by 5.3 mm. Maxillary height, as described by the nasion-ANS distance was shorter in the VDW patients, closely approaching statistical significance in the age range 7 through 11 years. The lip soft tissue measurements showed significantly greater protrusion over several age ranges in the VDW patients. The longitudinal growth analysis showed a significantly more inferior vertical position of the B point in the controls. CONCLUSIONS: This study demonstrates a few statistically significant differences in maxillary growth and lip conformation between VDW and matching controls. Small sample sizes for each age group hamper the ability to fully interpret or generalize the pattern of these differences. PMID- 11879082 TI - Modifying the two-stage cleft palate surgical correction. AB - OBJECTIVE: This paper reports the experience with a two-stage approach to surgical correction of the complete cleft palate, wherein timing of the second stage is dependent on the judgment of the speech pathologist and the orthodontist together with the surgeon. PATIENTS: Of a total of 35 patients having complete unilateral clefts a sample of 22 were available for postsurgical assessment. The first-stage repair of the palate was carried out at an average age of 10.7 months (range 6 to 17 months), and the second-stage repair of the residual cleft was completed at an average age of 32.7 months (range 26 to 34 months). INTERVENTIONS: The first-stage repair of the soft palate defect involved mobilizing two short posteriorly based flaps, which extend onto the posterior quarter of the hard palate thus including up to 1 cm of mucoperiosteum. Careful freeing of the muscle is followed by an intravelar veloplasty. The later closure of the residual cleft involved turnover hinge flaps and small mucoperiosteal flaps. RESULTS: Eighty-seven percent of the sample had good to excellent speech as assessed by the Great Ormond Street screening method. Only two patients showed evidence of recessive maxillae with Class III malocclusions. CONCLUSIONS: A two stage surgical closure of the palate using this procedure would appear to confer several valuable advantages to the patient. These include favorable outcomes for speech in the large majority of cases and minimal adverse effects on the growth of the midface region. PMID- 11879083 TI - Craniofacial and dental manifestations of Proteus syndrome: a case report. AB - The Proteus syndrome is a rare congenital hamartomatous condition that is characterized by a wide range of malformations, sometimes involving the face. Common manifestations include partial gigantism, congenital lipomas, and plantar hyperplasia. In this report we describe the craniofacial clinicopathological development in a girl with Proteus syndrome from age 6 to 20 years. The patient had pronounced hemifacial hypertrophy, exostoses in the left parietal region, and enlargement of the inferior alveolar nerve and mandibular canal in the affected region. The dental development of the affected left mandible and maxilla was characterized by extremely premature development and eruption of the primary and permanent teeth and by pronounced idiopathic root resorptions. The multidisciplinary management of the patient and the treatment outcome is reported. A review of the Proteus patients in the literature who exhibited manifestation in the craniofacial region is presented. PMID- 11879087 TI - Medical educators weigh curriculum changes to address threats of terrorism. PMID- 11879088 TI - Unusual stroke risk factors reported at ASA conference. PMID- 11879084 TI - Clinical and biochemical findings of a patient with thanatophoric dysplasia type I: additional finding of dicarboxylic aciduria. AB - OBJECTIVE: A long-surviving thanatophoric dysplasia type I patient to age of 6 years is presented. RESULTS AND CONCLUSIONS: Molecular studies revealed a heterozygous point mutation, S249C in the fibroblast growth factor receptor 3 gene. Most of the clinical course was similar to previous reports, including hearing loss and acanthosis nigricans. Abnormal urinary excretion of dicarboxylic acids and 3-hydroxydicarboxylic acids was observed. We hypothesize that this was a consequence of the FGFR3 mutation. PMID- 11879089 TI - Education needed to expand SCHIP eligibility. PMID- 11879090 TI - From the Food and Drug Administration. PMID- 11879094 TI - From the World Health Organization. PMID- 11879095 TI - Identifying fracture risk in postmenopausal women. PMID- 11879096 TI - Identifying fracture risk in postmenopausal women. PMID- 11879098 TI - Breastfeeding vs formula-feeding among HIV-infected women in resource-poor areas. PMID- 11879099 TI - Breastfeeding vs formula-feeding among HIV-infected women in resource-poor areas. PMID- 11879100 TI - Breastfeeding vs formula-feeding among HIV-infected women in resource-poor areas. PMID- 11879103 TI - Physicians' feelings about themselves and their patients. PMID- 11879104 TI - Physicians' feelings about themselves and their patients. PMID- 11879105 TI - Physicians' feelings about themselves and their patients. PMID- 11879107 TI - Molecular analysis of early postvaccine mumps-like disease in Italian military recruits. PMID- 11879108 TI - Effect of nonoxynol-9 gel on urogenital gonorrhea and chlamydial infection: a randomized controlled trial. AB - CONTEXT: Nonoxynol-9 has been suggested as a vaginal microbicide to protect against common sexually transmitted infections. OBJECTIVE: To compare nonoxynol-9 gel and condom use (gel group) vs condom use alone (condom group) for the prevention of male-to-female transmission of urogenital gonococcal and chlamydial infection. DESIGN AND SETTING: Randomized controlled trial conducted at 10 community clinics and 10 pharmacies in Yaounde, Cameroon, between October 1998 and September 2000, with 6 months of follow-up. PARTICIPANTS: High-risk population of 1251 women (excluding sex workers) being treated for or who had symptoms of sexually transmitted infections. Three were excluded from the gel group (0.5%) and 7 from the condom group (1%) because of no follow-up data. INTERVENTIONS: Nonoxynol-9 gel (100 mg) and condoms or condoms only. MAIN OUTCOME MEASURE: A positive test result for gonococcal or chlamydial infection by the ligase chain reaction assay; secondary outcome measure was a positive test result for human immunodeficiency virus (HIV). RESULTS: The rate ratio (RR) for new urogenital infections was 1.2 for the gel group vs condom group (95% confidence interval [CI], 0.9-1.6; P =.21). The gel group had 116 diagnosed gonococcal infections, chlamydial infections, or both for a rate of 43.6 per 100 person years, and the condom group had 100 infections for a rate of 36.6 per 100 person years. The RR for gonococcal infection in the gel group vs the condom group was 1.5 (95% CI, 1.0-2.3) and for chlamydial infection was 1.0 (95% CI, 0.7-1.4). There were 5 new cases of HIV infections in the gel group and 4 in the condom group. Three women in each group became pregnant during the study. CONCLUSION: Nonoxynol-9 gel did not protect against urogenital gonococcal or chlamydial infection. PMID- 11879109 TI - Cognitive functioning of long-term heavy cannabis users seeking treatment. AB - CONTEXT: Cognitive impairments are associated with long-term cannabis use, but the parameters of use that contribute to impairments and the nature and endurance of cognitive dysfunction remain uncertain. OBJECTIVE: To examine the effects of duration of cannabis use on specific areas of cognitive functioning among users seeking treatment for cannabis dependence. DESIGN, SETTING, AND PARTICIPANTS: Multisite retrospective cross-sectional neuropsychological study conducted in the United States (Seattle, Wash; Farmington, Conn; and Miami, Fla) between 1997 and 2000 among 102 near-daily cannabis users (51 long-term users: mean, 23.9 years of use; 51 shorter-term users: mean, 10.2 years of use) compared with 33 nonuser controls. MAIN OUTCOME MEASURES: Measures from 9 standard neuropsychological tests that assessed attention, memory, and executive functioning, and were administered prior to entry to a treatment program and following a median 17-hour abstinence. RESULTS: Long-term cannabis users performed significantly less well than shorter-term users and controls on tests of memory and attention. On the Rey Auditory Verbal Learning Test, long-term users recalled significantly fewer words than either shorter-term users (P =.001) or controls (P =.005); there was no difference between shorter-term users and controls. Long-term users showed impaired learning (P =.007), retention (P =.003), and retrieval (P =.002) compared with controls. Both user groups performed poorly on a time estimation task (P<.001 vs controls). Performance measures often correlated significantly with the duration of cannabis use, being worse with increasing years of use, but were unrelated to withdrawal symptoms and persisted after controlling for recent cannabis use and other drug use. CONCLUSIONS: These results confirm that long term heavy cannabis users show impairments in memory and attention that endure beyond the period of intoxication and worsen with increasing years of regular cannabis use. PMID- 11879111 TI - Clinical profile and spectrum of commotio cordis. AB - CONTEXT: Although blunt, nonpenetrating chest blows causing sudden cardiac death (commotio cordis) are often associated with competitive sports, dangers implicit in such blows can extend into many other life activities. OBJECTIVE: To describe the comprehensive spectrum of commotio cordis events. DESIGN AND SETTING: Analysis of confirmed cases from the general community assembled in the US Commotio Cordis Registry occurring up to September 1, 2001. MAIN OUTCOME MEASURE: Commotio cordis event. RESULTS: Of 128 confirmed cases, 122 (95%) were in males and the mean (SD) age was 13.6 (8.2) years (median, 14 years; range, 3 months to 45 years); only 28 (22%) cases were aged 18 years or older. Commotio cordis events occurred most commonly during organized sporting events (79 [62%]), such as baseball, but 49 (38%) occurred as part of daily routine and recreational activities. Fatal blows were inflicted with a wide range of velocities but often occurred inadvertently and under circumstances not usually associated with risk for sudden death in informal settings near the home or playground. Twenty-two (28%) participants were wearing commercially available chest barriers, including 7 in whom the projectile made direct contact with protective padding (baseball catchers and lacrosse/hockey goalies), and 2 in whom the projectile was a baseball specifically designed to reduce risk. Only 21 (16%) individuals survived their event, with particularly prompt cardiopulmonary resuscitation/defibrillation (most commonly reversing ventricular fibrillation) the only identifiable factor associated with a favorable outcome. CONCLUSIONS: The expanded spectrum of commotio cordis illustrates the potential dangers implicit in striking the chest, regardless of the intent or force of the blow. These findings also suggest that the safety of young athletes will be enhanced by developing more effective preventive strategies (such as chest wall barriers) to achieve protection from ventricular fibrillation following precordial blows. PMID- 11879110 TI - Lung cancer, cardiopulmonary mortality, and long-term exposure to fine particulate air pollution. AB - CONTEXT: Associations have been found between day-to-day particulate air pollution and increased risk of various adverse health outcomes, including cardiopulmonary mortality. However, studies of health effects of long-term particulate air pollution have been less conclusive. OBJECTIVE: To assess the relationship between long-term exposure to fine particulate air pollution and all cause, lung cancer, and cardiopulmonary mortality. DESIGN, SETTING, AND PARTICIPANTS: Vital status and cause of death data were collected by the American Cancer Society as part of the Cancer Prevention II study, an ongoing prospective mortality study, which enrolled approximately 1.2 million adults in 1982. Participants completed a questionnaire detailing individual risk factor data (age, sex, race, weight, height, smoking history, education, marital status, diet, alcohol consumption, and occupational exposures). The risk factor data for approximately 500 000 adults were linked with air pollution data for metropolitan areas throughout the United States and combined with vital status and cause of death data through December 31, 1998. MAIN OUTCOME MEASURE: All-cause, lung cancer, and cardiopulmonary mortality. RESULTS: Fine particulate and sulfur oxide -related pollution were associated with all-cause, lung cancer, and cardiopulmonary mortality. Each 10-microg/m(3) elevation in fine particulate air pollution was associated with approximately a 4%, 6%, and 8% increased risk of all-cause, cardiopulmonary, and lung cancer mortality, respectively. Measures of coarse particle fraction and total suspended particles were not consistently associated with mortality. CONCLUSION: Long-term exposure to combustion-related fine particulate air pollution is an important environmental risk factor for cardiopulmonary and lung cancer mortality. PMID- 11879112 TI - Risk of injury to child passengers in compact extended-cab pickup trucks. AB - CONTEXT: An increasing number of compact pickup trucks can accommodate restrained rear occupants. Rear seats in these pickup trucks are exempt from regulatory safety testing though their relative safety has not been determined. OBJECTIVES: To evaluate the risk of injury to children in compact extended-cab pickup trucks compared with children in other vehicles and to determine if any unique hazards exist. DESIGN: Cross-sectional study of children aged 15 years or younger in crashes of insured vehicles, with data collected via insurance claim records and a telephone survey. SETTING AND PARTICIPANTS: Probability sample of 7192 multirow vehicles involved in crashes, with 11 335 child occupants, in 3 large US regions from December 1, 1998, to November 30, 2000. MAIN OUTCOME MEASURE: Relative risk of injury, defined as concussions and more serious brain injuries, spinal cord injuries, internal organ injuries, extremity fractures, and facial lacerations, estimated by odds ratios (ORs) adjusting for age, restraint use, point of impact, vehicle weight, and crash severity. RESULTS: Injuries were reported for 1356 children, representing 1.6% of the population. Children in compact extended-cab pickup trucks were at greater risk of injury than children in other vehicles (adjusted OR, 2.96; 95% confidence interval [CI], 1.68-5.21). Children in the rear seats of compact pickup trucks were at substantially greater risk of injury than rear-seated children in other vehicles (adjusted OR, 4.75; 95% CI, 2.39 9.43). Children seated in the front seat of compact extended-cab pickup trucks were at greater risk of injury than children in the front seats of other vehicles, but this risk was not statistically significant (adjusted OR, 1.70; 95% CI, 0.78-3.69). CONCLUSIONS: Children in compact extended-cab pickup trucks are not as safe as children in other vehicles, primarily due to the increased relative risk of injury in the back seat. For families with another choice of vehicle, clinicians should advise parents against transporting children in compact pickup trucks. The current exemption for regulatory testing for occupant protection in the rear seats of compact pickup trucks should be reconsidered. PMID- 11879113 TI - Retinal arteriolar narrowing and risk of coronary heart disease in men and women. The Atherosclerosis Risk in Communities Study. AB - CONTEXT: Microvascular processes have been hypothesized to play a greater role in the development of coronary heart disease (CHD) in women than in men; however, prospective clinical data are limited. OBJECTIVE: To examine the association between retinal arteriolar narrowing, a marker of microvascular damage from hypertension and inflammation, and incident CHD in healthy middle-aged women and men. DESIGN, SETTING, AND PARTICIPANTS: The Atherosclerosis Risk in Communities Study, an ongoing prospective, population-based cohort study in 4 US communities initiated in 1987-1989. Retinal photographs were taken in 9648 women and men aged 51 to 72 years without CHD at the third examination (1993-1995). To quantify retinal arteriolar narrowing, the photographs were digitized, individual arteriolar and venular diameters were measured, and a summary arteriole-to-venule ratio (AVR) was calculated. MAIN OUTCOME MEASURE: Risk of CHD associated with retinal arteriolar narrowing. RESULTS: During an average 3.5 years of follow-up, 84 women and 187 men experienced incident CHD events. In women, after controlling for mean arterial blood pressure averaged over the previous 6 years, diabetes, cigarette smoking, plasma lipid levels, and other risk factors, each SD decrease in the AVR was associated with an increased risk of any incident CHD (relative risk [RR], 1.37; 95% confidence interval [CI], 1.08-1.72) and of acute myocardial infarction (RR, 1.50; 95% CI, 1.10-2.04). In contrast, AVR was unrelated to any incident CHD in men (RR, 1.00; 95% CI, 0.84-1.18) or to acute myocardial infarction (RR, 1.08; 95% CI, 0.85-1.38). CONCLUSION: Retinal arteriolar narrowing is related to risk of CHD in women but not in men, supporting a more prominent microvascular role in the development of CHD in women than in men. Future work is needed to confirm these findings. PMID- 11879114 TI - Is this patient clinically depressed? AB - CONTEXT: Depressive disorders are highly prevalent in the general population, but recognition and accurate diagnosis are made difficult by the lack of a simple confirmatory test. OBJECTIVE: To review the accuracy and precision of depression questionnaires and the clinical examination for diagnosing clinical depression. DATA SOURCES: We searched the English-language literature from 1970 through July 2000 using MEDLINE, a specialized registry of depression trials, and bibliographies of selected articles. STUDY SELECTION: Case-finding studies were included if they used depression questionnaires with easy to average literacy requirements, evaluated at least 100 primary care patients, and compared questionnaire results with accepted diagnostic criteria for major depression. Eleven questionnaires, ranging in length from 1 to 30 questions, were assessed in 28 published studies. Reliability studies for the clinical examination required criterion-based diagnoses made by at least 2 clinicians who interviewed the patient or reviewed a taped examination. Fourteen studies evaluated interrater reliability. DATA EXTRACTION: Pairs of authors independently reviewed articles. For case-finding studies, quality assessment addressed sample size and whether patients were selected consecutively or randomly, the criterion standard was administered and interpreted independently of and blind to the results of the case-finding instrument, and the proportion of persons receiving the criterion standard assessment was less than or more than 50% of those approached for criterion standard assessment. For reliability studies, quality assessment addressed whether key patient characteristics were described, the interviewers collected clinical history independently, and diagnoses were made blinded to other clinicians' evaluations. DATA SYNTHESIS: In case-finding studies, average questionnaire administration times ranged from less than 1 minute to 5 minutes. The median likelihood ratio positive for major depression was 3.3 (range, 2.3 12.2) and the median likelihood ratio negative was 0.19 (range, 0.14-0.35). No significant differences between questionnaires were found. For mental health care professionals using a semistructured interview, agreement was substantial to almost perfect for major depression (kappa = 0.64-0.93). Nonstandardized interviews yielded somewhat lower agreement (kappa = 0.55-0.74). A single study showed that primary care clinicians using a semistructured interview have high agreement with mental health care professionals (kappa = 0.71). CONCLUSIONS: Multiple, practical questionnaires with reasonable performance characteristics are available to help clinicians identify and diagnose patients with major depression. Diagnostic confirmation by mental health care professionals using a clinical interview or by primary care physicians using a semistructured interview can be made with high reliability. PMID- 11879115 TI - Nonoxynol-9 as a vaginal microbicide for prevention of sexually transmitted infections: it's time to move on. PMID- 11879116 TI - Cannabis, cognition, and residual confounding. PMID- 11879118 TI - MSJAMA. Dissecting Gross Anatomy. PMID- 11879119 TI - MSJAMA. From theater to laboratory. PMID- 11879120 TI - MSJAMA. The changing role of dissection in medical education. PMID- 11879121 TI - MSJAMA. Art, anatomy, learning, and living. PMID- 11879122 TI - MSJAMA. From grave robbing to gifting: cadaver supply in the United States. PMID- 11879129 TI - A randomized trial of atropine vs. patching for treatment of moderate amblyopia in children. AB - OBJECTIVE: To compare patching and atropine as treatments for moderate amblyopia in children younger than 7 years. METHODS: In a randomized clinical trial, 419 children younger than 7 years with amblyopia and visual acuity in the range of 20/40 to 20/100 were assigned to receive either patching or atropine at 47 clinical sites. MAIN OUTCOME MEASURE: Visual acuity in the amblyopic eye and sound eye after 6 months. RESULTS: Visual acuity in the amblyopic eye improved in both groups (improvement from baseline to 6 months was 3.16 lines in the patching group and 2.84 lines in the atropine group). Improvement was initially faster in the patching group, but after 6 months, the difference in visual acuity between treatment groups was small and clinically inconsequential (mean difference at 6 months, 0.034 logMAR units; 95% confidence interval, 0.005-0.064 logMAR units). The 6-month acuity was 20/30 or better in the amblyopic eye and/or improved from baseline by 3 or more lines in 79% of the patching group and 74% of the atropine group. Both treatments were well tolerated, although atropine had a slightly higher degree of acceptability on a parental questionnaire. More patients in the atropine group than in the patching group had reduced acuity in the sound eye at 6 months, but this did not persist with further follow-up. CONCLUSION: Atropine and patching produce improvement of similar magnitude, and both are appropriate modalities for the initial treatment of moderate amblyopia in children aged 3 to less than 7 years. PMID- 11879130 TI - The clinical profile of moderate amblyopia in children younger than 7 years. AB - OBJECTIVE: To describe the demographic and clinical characteristics of a cohort of children with moderate amblyopia participating in the Amblyopia Treatment Study 1, a randomized trial comparing atropine and patching. METHODS: The children enrolled were younger than 7 years and had strabismic, anisometropic, or combined strabismic and anisometropic amblyopia. Visual acuity, measured with a standardized testing protocol using single-surround HOTV optotypes, was 20/40 to 20/100 in the amblyopic eye, with an intereye acuity difference of 3 or more logMAR lines. There were 419 children enrolled, 409 of whom met these criteria and were included in the analyses. RESULTS: The mean age of the 409 children was 5.3 years. The cause of the amblyopia was strabismus in 38%, anisometropia in 37%, and both strabismus and anisometropia in 24%. The mean visual acuity of the amblyopic eyes (approximately 20/60) was similar among the strabismic, anisometropic, and combined groups (P =.24), but visual acuity of the sound eyes was worse in the strabismic group compared with the anisometropic group (P<.001). For the patients randomized into the patching group, 43% were initially treated for 6 hours per day, whereas 17% underwent full-time patching. Patients with poorer visual acuity in the amblyopic eye were prescribed more hours of patching than patients with better acuity (P =.003). CONCLUSIONS: In the Amblyopia Treatment Study 1, there were nearly equal proportions of patients with strabismic and anisometropic amblyopia. A similar level of visual impairment was found irrespective of the cause of amblyopia. There was considerable variation in treatment practices with regard to the number of hours of initial patching prescribed. PMID- 11879131 TI - Histologic and ultrastructural findings in human corneas after successful laser in situ keratomileusis. AB - OBJECTIVE: To examine the histologic and ultrastructural features of human corneas after successful laser in situ keratomileusis (LASIK) in 2 patients post mortem. METHODS: Portions of 4 corneas were processed for histology, transmission electron microscopy, and scanning electron microscopy. RESULTS: Case 1 had undergone LASIK 3 months prior to death and case 2 had undergone LASIK 20 months prior to death. A Hansatome (Bausch & Lomb Surgical Inc, Clarement, Calif) microkeratome with a 180-microm plate had been used for case 1 and an Automated Corneal Shaper (Chiron Corporation, Munich, Germany) with a 160-microm plate had been used for case 2. Histologically, the LASIK flap measured 160 microm and 150 microm in thickness in case 1 and case 2, respectively. Corneas from both cases exhibited minor epithelial ingrowth into the wound, reactive keratocytes at the wound margin, irregular collagen fibrils in the wound bed, and severed collagen bundles at the flap hinge. These findings were more pronounced in case 1 than in case 2, and the wound interface was virtually imperceptible in case 2. Additionally, the corneas from case 1 contained periodic acid-Schiff--positive electron dense material and wide-spaced collagen at the wound interface, and there was an absence of corneal nerves. CONCLUSIONS: These findings show that changes caused by wound repair that are present at 3 months are minor 20 months after LASIK. PMID- 11879132 TI - Incidence of late-onset bleb-related complications following trabeculectomy with mitomycin. AB - OBJECTIVES: To determine the incidence of late-onset bleb-related complications following trabeculectomy with mitomycin and to report the management and outcome of bleb leaks following trabeculectomy with mitomycin. METHODS: A retrospective medical record review of all patients who underwent trabeculectomy with mitomycin from June 1, 1991, through April 30, 1998, at our institution was performed. The Kaplan-Meier survival method was used to estimate the probability of (1) endophthalmitis, (2) blebitis, (3) a bleb leak, and (4) the combined outcome (the first occurrence of a bleb leak, blebitis, or endophthalmitis). This survival analysis included only the first trabeculectomy in an eye, with at least 3 months of follow-up during the study period. A separate description of bleb leak management and outcome was performed. RESULTS: Two hundred thirty-nine eyes of 198 patients were included in the survival analysis. The average follow-up was 2.7 (range, 0.3-7.3) years. Twenty eyes (8%) from 19 patients experienced a bleb leak; the adjusted incidence was 3.2% per patient-year. Five eyes (2%) had an episode of blebitis. Eight eyes (3%) experienced an episode of endophthalmitis; the follow-up adjusted incidence (number of events per patient-year) was 1.3%. Twenty-seven eyes (11%) from 26 patients had at least 1 of the complications of a bleb leak, blebitis, or endophthalmitis; the adjusted incidence was 4.4% per patient-year. A Kaplan-Meier analysis estimated the 5-year probability of developing a bleb leak, blebitis, or endophthalmitis to be 17.9%, 6.3%, and 7.5%, respectively. Two hundred fifty-eight trabeculectomies in 242 eyes of 198 patients were included in the description of bleb leak management and outcome. Bleb leaks occurred in 22 eyes (9% of the 258 trabeculectomies). Seventeen eyes were successfully treated with office-based measures, and 4 ultimately underwent surgical bleb revision. One eye without infection continued to leak after 11 months of office-based therapy. CONCLUSIONS: There is significant morbidity associated with a trabeculectomy with mitomycin. The incidence of a bleb leak or an infection continues at a fairly constant rate over time, such that at 5 years, up to 23% of all patients might develop one of these complications. An isolated bleb leak seems to be a relatively benign condition, as three quarters resolve with office-based methods. PMID- 11879133 TI - Vascular dysregulation in the choroid of subjects with acral vasospasm. AB - OBJECTIVE: To assess the relationship between ocular perfusion pressure and blood flow in the choroid in subjects with acral vasospasm. PATIENTS AND METHODS: Twenty otherwise healthy subjects with acral vascular dysregulation and 55 age matched nonvasospastic healthy volunteers were recruited. After a 20-minute rest in a sitting position, intraocular pressure and choroidal blood flow were determined by means of applanation tonometry and choroidal laser Doppler flowmetry, respectively. The laser Doppler flowmetry variables velocity, volume, and flux were assessed. The correlations between mean ocular perfusion pressure ([23 x [(23 x diastolic blood pressure) + (13 x systolic blood pressure)]] intraocular pressure) and blood flow measures were determined by means of the Pearson linear correlation factor. The t test was used to evaluate differences between normal subjects and patients with vasospasm. RESULTS: Apart from a slight difference in systolic blood pressure (mean +/- SD, 113.70 +/- 11.88 mm Hg in the vasospastic group and 121.09 +/- 14.58 mm Hg in the control group; P =.05), the 2 study groups were completely comparable. Velocity and flux correlated significantly with the mean ocular perfusion pressure (r = 0.76, P<.001; r = 0.64, P =.002, respectively) in vasospastic subjects. Such correlations did not occur in the control group, and the difference between vasospastic patients and control subjects with regard to these correlations was statistically significant (P<.001 and P =.003, respectively). CONCLUSIONS: Choroidal blood flow seems, to some degree, to be independent of perfusion pressure, but not in subjects with acral vasospasm. PMID- 11879134 TI - Solitary idiopathic choroiditis: the Richard B. Weaver lecture. AB - OBJECTIVE: To report the clinical characteristics of solitary idiopathic choroiditis and the features that differentiate it from tumors and other inflammatory lesions. DESIGN: Retrospective medical record review. PATIENTS: Sixty consecutive patients with solitary idiopathic choroiditis. MAIN OUTCOME MEASURES: Clinical features, natural course, and follow-up. RESULTS: The mean and median ages of the patients were 35 and 36 years, respectively, 56 (93%) of the 60 patients were white, 38 (63%) were female, and 22 (37%) were male. No patient had a history, clinical findings, or laboratory study results to support a specific cause of uveitis. All were referred to us because of a suspected intraocular tumor. The patient was asymptomatic in 21 cases (35%); the patient had mild visual loss in 36 cases (60%). The lesion was posterior to the equator in 56 cases (93%), was yellow in 58 cases (97%), and had distinct margins in 38 cases (63%). The lesion showed signs of active inflammation in 20 cases (33%) and no inflammation in 40 cases (67%). Fluorescein angiography disclosed that all lesions had early hypofluorescence and late hyperfluorescence. In the late-phase angiograms inactive lesions had distinct margins and active lesions had ill defined margins. Lesions with active inflammation appeared to show a favorable response to the administration of systemic corticosteroids but generally improved with or without treatment. Most of the inactive lesions remained stable on long term follow-up. Overall, the condition remained stable in 36 patients (60%), improved in 22 (37%), and recurred in 2 (3%). The clinical and angiographic features and clinical course of solitary idiopathic choroiditis were generally different from known intraocular tumors. CONCLUSIONS: Solitary idiopathic choroiditis is a distinct clinical entity that can simulate an intraocular neoplasm. Recognition of its typical clinical features can assist in differentiating it from tumors and other inflammatory lesions of the ocular fundus. PMID- 11879135 TI - Neovascular membranes associated with idiopathic juxtafoveolar telangiectasis. AB - OBJECTIVE: To report the visual outcome in patients with a neovascular membrane (NVM) associated with idiopathic juxtafoveolar telangiectasis (IJFT). METHODS: We performed a retrospective, noncomparative analysis of 26 eyes of 16 patients with an NVM associated with bilateral IJFT (Gass classification group 2A). Eyes were divided into 2 groups: group WO (n = 11) included eyes with IJFT without evidence of an NVM on initial examination; eyes in group W (n = 15) had an NVM at the initial diagnosis of IJFT. In group WO, the initial visual acuity and the time between the initial examination to the diagnosis of an NVM were evaluated. Characteristic fundus findings, including the presence or absence of a chorioretinal anastomosis, intraretinal pigmentary plaques, and crystalline deposits, as well as the final visual acuity were reviewed for both groups. RESULTS: The initial visual acuity for eyes in group WO ranged from 20/20 to 20/70 (median, 20/30); in group W, from 20/20 to 4/200 (median, 20/70). The average time from initial diagnosis of IJFT to the development of an NVM was 73 months (range, 5-142 months). In group WO, chorioretinal anastomosis and concurrent perivascular retinal pigment epithelial hyperplasia were observed before the development of an NVM. The final visual acuity for all eyes ranged from 20/40 to 2/200 (median, 20/200). Eighty-one percent of eyes (21/26) had a final visual acuity of 20/200 or worse. CONCLUSIONS: The stable final visual acuity in patients with an NVM associated with IJFT is generally poor, with 80% of eyes in this series having a final visual acuity of l20/200 or worse. In patients with IJFT, the presence of a chorioretinal anastomosis and retinal pigment epithelial hyperplastic plaques always preceded the development of an NVM. PMID- 11879136 TI - Twenty-year follow-up for scleral buckling. AB - OBJECTIVE: To the report 20-year follow-up data for patients receiving a scleral buckle for treatment of a primary rhegmatogenous retinal detachment (RRD). METHODS: Nonconsecutive, retrospective case series. We identified 227 eyes with primary RRD who were treated with a scleral buckle, and for whom at least 20 years of follow-up data were available. Results were classified into 3 subgroups: retina reattached with 1 procedure; retina reattached with 1 or more additional vitreoretinal procedures; or retina detached at 20 years. RESULTS: One hundred eighty-six eyes (82%) achieved retinal reattachment with 1 scleral buckling procedure and with a median final visual acuity of 20/40 at 20 years of follow up. An additional 30 eyes (13%) achieved retinal reattachment after 1 or more additional vitreoretinal procedures, with a median final visual acuity of 20/50. Eleven eyes (5%) were detached at the 20-year follow-up examination, with a final visual acuity in all eyes of no light perception. CONCLUSIONS: Scleral buckling for primary RRD achieves anatomical efficacy and preservation of central vision in the majority of eyes at 20 years' follow-up. The 1-operation success rate was 82%, overall success rate was 95%, and median final visual acuity was 20/40. This study may serve as a basis for comparison with the long-term results of other surgical techniques used in the treatment of primary RRDs. PMID- 11879137 TI - Goblet cell numbers and epithelial proliferation in the conjunctiva of patients with dry eye syndrome treated with cyclosporine. AB - OBJECTIVES: To compare conjunctival goblet cell numbers as well as epithelial turnover in patients with non-Sjogren syndrome--associated keratoconjunctivitis sicca (NSS-KCS) and those with SS-KCS before and after 6 months of treatment with topical cyclosporine A (CsA) ophthalmic emulsion. METHODS: Conjunctival biopsy specimens from 16 patients with NSS-KCS and 12 with SS-KCS were obtained at baseline and after 6 months' therapy with CsA or vehicle alone. Conjunctival biopsy specimens were also obtained from 11 normal subjects. Periodic acid- Schiff staining determined the number of goblet cells present. Immunofluorescence microscopy for Ki-67 localization was used to evaluate the number of actively cycling cells. RESULTS: Periodic acid--Schiff staining showed fewer goblet cells at baseline in both dry eye populations when compared with normal subjects (P<.001). After 6 months of CsA treatment, conjunctival biopsy specimens of both NSS-KCS and SS-KCS groups revealed an increase in goblet cells compared with baseline (P<.05). More Ki-67--positive cells were observed in NSS-KCS conjunctiva at baseline than in normal conjunctiva (P<.05) whereas numbers of these cells in SS-KCS conjunctiva were similar to normal at baseline. After 6 months of CsA treatment, conjunctival biopsy specimens of NSS-KCS revealed a decrease in Ki-67- labeled cells compared with baseline (P<.001). In contrast, no substantial change was observed for CsA treatment in patients with SS-KCS. CONCLUSIONS: Treatment of dry eye syndrome for 6 months with topical CsA resulted in an increase in goblet cell numbers in patients with NSS-KCS and SS-KCS and a decrease in epithelial turnover in those with NSS-KCS. Reducing ocular surface inflammation might have an effect on the proliferative activity of the epithelium. PMID- 11879138 TI - Prevention of experimental choroidal neovascularization with intravitreal anti vascular endothelial growth factor antibody fragment. AB - OBJECTIVE: To evaluate the safety and efficacy of intravitreal injections of an antigen-binding fragment of a recombinant humanized monoclonal antibody directed toward vascular endothelial growth factor (rhuFab VEGF) in a monkey model of choroidal neovascularization (CNV). METHODS: In phase 1 of the study, each animal received intravitreal injections, 500 microg per eye, of rhuFab VEGF in one eye (prevention eye), while the contralateral eye received rhuFab VEGF vehicle (control eye) at 2-week intervals. On day 21, laser photocoagulation was performed to induce CNV. In phase 2, the vehicle-treated eye was crossed over and both eyes received 500 microg of rhuFab VEGF beginning 21 days following laser induced injury at days 42 and 56. The eyes were monitored by ophthalmic examinations, color photographs, and fluorescein angiography. RESULTS: rhuFab VEGF did not cause any ocular hemorrhages. All eyes treated with rhuFab VEGF developed acute anterior chamber inflammation within 24 hours of the first injection that resolved within 1 week, and this inflammation was less severe with subsequent injections. The incidence of CNV, defined angiographically, was significantly lower in the prevention eyes than the control eyes (P<.001). Subsequent treatments were associated with less leakage in eyes with established CNV that were crossed over from the control eyes to the treatment eyes (P =.001). CONCLUSIONS: Intravitreal rhuFab VEGF injections prevented formation of clinically significant CNV in cynomolgus monkeys and decreased leakage of already formed CNV with no significant toxic effects. CLINICAL RELEVANCE: This study provides the nonclinical proof of principle for ongoing clinical studies of intravitreally injected rhuFab VEGF in patients with neovascular age-related macular degeneration. PMID- 11879139 TI - Partial antagonism of endothelin 1-induced vasoconstriction in the human choroid by topical unoprostone isopropyl. AB - BACKGROUND: There is increasing evidence that reduced ocular blood flow plays a role in the pathogenesis of glaucoma. In patients with normal-tension glaucoma, ocular blood flow abnormalities may be associated with dysfunction of the endothelin 1 (ET-1) regulation system. OBJECTIVE: To test the hypothesis that unoprostone, a topical docosanoid, may affect ET-1--induced vasoconstriction in the human choroid. METHODS: In a placebo-controlled, randomized, double-masked, 2 way crossover design, ET-1 (2.5 ng/kg per minute for 150 minutes) was administered intravenously to 24 healthy individuals. Thirty minutes after the start of ET-1 infusion, 1 drop of unoprostone or placebo was instilled into the right eye. After another 30 minutes, 2 drops of unoprostone or placebo was topically administered. This procedure was continued and the dose was increased further until 4 drops of unoprostone or placebo was reached. Subfoveal and pulsatile choroidal blood flow were assessed using laser Doppler flowmetry and laser interferometric measurement of fundus pulsation amplitude, respectively. RESULTS: Administration of exogenous ET-1 decreased choroidal blood flow (mean +/ SEM, 17% +/- 2%; P<.001) and fundus pulsation amplitude (mean +/- SEM, 19% +/- 2%; P<.001). This effect was significantly blunted when topical unoprostone was coadministered (mean +/- SEM decrease in choroidal blood flow, 7% +/- 2%; P =.04 vs. placebo; mean +/- SEM decrease in fundus pulsation amplitude, 12% +/- 2%; P<.001 vs. placebo). CONCLUSION: There is a functional antagonism between ET-1 and topical unoprostone in the choroidal vasculature. CLINICAL RELEVANCE: Our findings of a functional antagonism between ET-1 and topical unoprostone in the choroidal vasculature may be important in vascular eye diseases associated with increased ET-1. PMID- 11879140 TI - Hyperfractionated external beam radiation therapy in the treatment of murine transgenic retinoblastoma. AB - OBJECTIVE: To determine the in vivo efficacy of hyperfractionated external beam radiation therapy (EBRT) in comparison with standard daily EBRT in a murine model of heritable retinoblastoma. METHODS: Two hundred twenty eyes from 6-week-old simian virus-40 large T-antigen--positive mice were treated with a total dose of EBRT ranging from 10-76 Gy (1000 to 7600 rad). One hundred ten eyes underwent EBRT administered in 2.0-Gy (200-rad) fractions once per day. Forty-two eyes received hyperfractionated EBRT administered in 1.2-Gy (120-rad) fractions twice per day, while 48 eyes received EBRT twice daily in fractions of 5.0 Gy (500 rad). Twenty eyes served as untreated controls. All eyes were obtained for histopathologic examination and graded positive if any tumor was present. RESULTS: A dose-dependent inhibition of ocular tumor was observed for EBRT in these transgenic retinoblastoma mice. The tumor control dose for 50% of eyes (TCD(50)) treated with 2.0 Gy fractions of EBRT was 45 Gy (4500 rad) when treatments were administered once daily. A significant increase in tumor control was observed when treatments were administered twice per day at fractions of 1.2 Gy, resulting in a TCD(50) of 33 Gy (3300 rad) (P =.003). A further increase in tumor control was observed when twice-daily EBRT was administered in 5.0 Gy fractions resulting in a TCD(50) of 28 Gy (2800 rad). CONCLUSIONS: Hyperfractionated EBRT safely and effectively controls intraocular retinoblastoma in this transgenic animal model. Use of hyperfractionation allows for a reduction in total radiation delivered dose, while shortening the total treatment time. CLINICAL RELEVANCE: This treatment approach may be applicable in the management of pediatric retinoblastoma by maintaining excellent tumor control, while reducing treatment-associated complications. PMID- 11879141 TI - Adaptations and deficits in the vestibulo-ocular reflex after third nerve palsy. AB - OBJECTIVE: To analyze the vestibulo-ocular reflex (VOR) in patients with unilateral peripheral third nerve palsy. PARTICIPANTS AND METHODS: Ten patients and 15 healthy subjects were studied using magnetic search coils. Subjects made sinusoidal +/-10 degrees head-on-body rotations in yaw, pitch, and roll in darkness and during monocular viewing in light. RESULTS: Horizontal VOR and visually enhanced VOR (VVOR) gains of the paretic eye were decreased during both abduction and adduction. Vertical VOR and VVOR gains of the paretic eye were decreased during both elevation and depression. Dynamic and static torsional VOR and VVOR gains of the paretic eye were reduced during both excyclotorsion and incyclotorsion. Horizontal, vertical, and torsional VOR and VVOR gains were normal in the nonparetic eye. CONCLUSIONS: Adducting VOR gains were reduced as anticipated from medial rectus palsy. Abducting gains were also reduced; the reduction is attributed to an adaptive decrease in innervation to the lateral rectus to achieve symmetry of the horizontal VOR in the paretic eye. Torsional VOR gains were reduced during excyclotorsion from palsy of the inferior oblique muscle. Gains were also reduced during incyclotorsion, which can be explained by an adaptive decrease in innervation to the superior oblique to restore symmetry of the torsional VOR in the paretic eye. CLINICAL RELEVANCE: Monocular adaptation in the VOR of the paretic eye reduces asymmetrical movement of retinal images during head motion, prevents nystagmus, and reduces retinal image disparity. PMID- 11879142 TI - Novel mutations in the NRL gene and associated clinical findings in patients with dominant retinitis pigmentosa. AB - OBJECTIVES: To search for mutations in the neural retina leucine zipper (NRL) gene in patients with dominant retinitis pigmentosa and to compare the severity of disease in these patients with that observed previously in patients with dominant rhodopsin mutations. METHODS: Single-strand conformation analysis was used to survey 189 unrelated patients for mutations. The available relatives of index patients with mutations were also evaluated. In our clinical examination of patients, we measured visual acuity, final dark-adaptation threshold equivalent visual field diameter, and electroretinogram amplitudes among other parameters of visual function. We compared the clinical findings with those obtained earlier from similar evaluations of a group of 39 patients with the dominant rhodopsin mutation Pro23His and a group of 25 patients with the dominant rhodopsin mutation Pro347Leu. RESULTS: We identified 3 novel missense mutations in a total of 4 unrelated patients with dominant retinitis pigmentosa: Ser50Pro, Ser50Leu (2 patients), and Pro51Thr. Each mutation cosegregated with dominant retinitis pigmentosa. None of these mutations were found among 91 unrelated control individuals. The visual acuities among the 4 index patients and 3 relatives with NRL mutations who were clinically evaluated ranged from 20/20 (in a 9-year-old patient) to 20/200 (in a 73-year-old patient). All patients had bone-spicule pigment deposits in their fundi. Average rod-plus-cone and cone-isolated electroretinogram amplitudes were both decreased by 99% or more compared with normal amplitudes. The dark-adaptation thresholds, equivalent visual field diameters, and electroretinogram amplitudes (all corrected for age and refractive error) indicated that the disease caused by the NRL mutations was more severe than that caused by the dominant rhodopsin mutation Pro23His and was similar in severity to that produced by the rhodopsin mutation Pro347Leu. CONCLUSION: The 3 novel NRL mutations we discovered bring the total number of reported mutations in this gene to 6. Five of the 6 mutations affect residues 50 or 51, suggesting that these residues are important in a structural or functional domain of the encoded protein. CLINICAL RELEVANCE: Rod and cone function is affected to a similar degree in patients with these mutations. The disease caused by NRL mutations found in this study appears to be more severe than that caused by the rhodopsin mutation Pro23His and is similar in severity to that caused by the rhodopsin mutation Pro347Leu, even after correcting for age. PMID- 11879143 TI - Novel mutation in the TIMP3 gene causes Sorsby fundus dystrophy. AB - OBJECTIVE: To determine the molecular basis of a retinopathy previously described as dominant macular subretinal neovascularization with peripheral retinal degeneration. METHODS: The TIMP3 gene was analyzed in family members, and 4 mutation-positive patients were studied using psychophysics and electroretinography. RESULTS: Cosegregating with disease in the family was a single base pair change in the TIMP3 gene, altering a conserved tyrosine to cysteine at amino acid position 172 (Y172C). There was psychophysical and electroretinographic evidence of rod dysfunction greater than cone dysfunction. Dark adaptometry showed abnormalities with regional retinal variation in degree. CONCLUSIONS: The Y172C mutation in the TIMP3 gene is another cause of Sorsby fundus dystrophy. The expression of this form of the disease, as in other C terminal TIMP3 mutations, is speculated to be secondary to mutant TIMP-3, causing a decreased turnover of the extracellular matrix. CLINICAL RELEVANCE: The molecular clarification of inherited retinal degeneration involving abnormal extracellular matrix turnover in and around Bruch's membrane should provide clues to the pathogenesis of not only these particular diseases but also forms of age related macular degeneration. PMID- 11879144 TI - Insights into the pathogenesis of thyroid-associated orbitopathy: evolving rationale for therapy. PMID- 11879145 TI - Atropine vs. patching for the treatment of moderate amblyopia in children. PMID- 11879146 TI - Translational research in ophthalmology. PMID- 11879147 TI - Calcium precipitation on the optical surfaces of a foldable intraocular lens: a clinicopathological correlation. PMID- 11879148 TI - Choroidal melanoma treated with cryotherapy. PMID- 11879149 TI - The World Trade Center disaster: a brief on-site report from Ground Zero. PMID- 11879150 TI - Regression of Elschnig pearls on the posterior capsule in a pseudophakic eye. PMID- 11879151 TI - Repair of impending macular hole in the early postoperative period as evaluated by optical coherence tomography. PMID- 11879152 TI - Cryotherapy as a primary treatment for choroidal melanoma. PMID- 11879153 TI - Photo essay: iris-lens inclusion cyst following penetrating injury. PMID- 11879154 TI - How big is small? PMID- 11879156 TI - Association between light exposure during the night and myopia in children. PMID- 11879157 TI - Ruptured globe caused by a fishing sinker. PMID- 11879158 TI - Differences in early childhood risk factors for juvenile-onset and adult-onset depression. AB - BACKGROUND: Family and twin studies suggest that juvenile-onset major depressive disorder (MDD) may be etiologically distinct from adult-onset MDD. This study is the first to distinguish prospectively between juvenile- and adult-onset cases of MDD in a representative birth cohort followed up from childhood into adulthood. METHOD: The study followed a representative birth cohort prospectively from birth to age 26 years. Early childhood risk factors covered the period from birth to age 9 years. Diagnoses of MDD were made according to DSM criteria at 3 points prior to adulthood (ages 11, 13, and 15 years) and 3 points during adulthood (ages 18, 21, and 26 years). Four groups were defined as (1) individuals first diagnosed as having MDD in childhood, but not in adulthood (n = 21); (2) individuals first diagnosed as having MDD in adulthood (n = 314); (3) individuals first diagnosed in childhood whose depression recurred in adulthood by age 26 years (n = 34); and (4) never-depressed individuals (n = 629). RESULTS: The 2 juvenile-onset groups had similar high-risk profiles on the childhood measures. Compared with the adult-depressed group, the juvenile-onset groups experienced more perinatal insults and motor skill deficits, caretaker instability, criminality, and psychopathology in their family-of-origin, and behavioral and socioemotional problems. The adult-onset group's risk profile was similar to that of the never-depressed group with the exception of elevated childhood sexual abuse. CONCLUSIONS: Heterogeneity within groups of psychiatric patients poses problems for theory, research, and treatment. The present study illustrates that the distinction between juvenile vs adult-onset MDD is important for understanding heterogeneity within depression. PMID- 11879159 TI - Juvenile-onset major depression includes childhood- and adolescent-onset depression and may be heterogeneous. PMID- 11879160 TI - Mental health, educational, and social role outcomes of adolescents with depression. AB - BACKGROUND: This study used longitudinal data to examine the extent to which young people with depression in mid adolescence (ages 14-16) were at increased risk of adverse psychosocial outcomes in later adolescence and young adulthood (ages 16-21). METHODS: Data were gathered during a 21-year longitudinal study of a birth cohort of 1265 children. Measures included assessments of DSM-III-R major depression (at age 14-16); psychiatric disorders, educational achievement, and social functioning (at age 16-21); social, familial, and individual factors; and comorbid disorders. RESULTS: Thirteen percent of the cohort developed depression between ages 14 and 16. Young people with depression in adolescence were at significantly (P<.05) increased risk of later major depression, anxiety disorders, nicotine dependence, alcohol abuse or dependence, suicide attempt, educational underachievement, unemployment, and early parenthood. These associations were similar for girls and boys. The results suggested the presence of 2 major pathways linking early depression to later outcomes. First, there was a direct linkage between early depression and increased risk of later major depression or anxiety disorders. Second, the associations between early depression and other outcomes were explained by the presence of confounding social, familial, and individual factors. CONCLUSIONS: Young people having early depression were at increased risk of later adverse psychosocial outcomes. There was a direct linkage in which early depression was associated with increased risk of later major depression and anxiety disorders. Linkages between early depression and other outcomes appeared to reflect the effects of confounding factors. PMID- 11879161 TI - Double-blind switch study of imipramine or sertraline treatment of antidepressant resistant chronic depression. AB - BACKGROUND: Although various strategies have been proposed to treat antidepressant nonresponders, little controlled research has been published that examines prospectively the use of switching to an alternate antidepressant. METHODS: This was a multisite study in which outpatients with chronic major depression (with or without concurrent dysthymia), who failed to respond to 12 weeks of double-blind treatment with either sertraline hydrochloride (n = 117) or imipramine hydrochloride (n = 51), were crossed over or switched to 12 additional weeks of double-blind treatment with the alternate medication. Outcome measures included the 24-item Hamilton Rating Scale for Depression and the Clinical Global Impressions--Severity and Improvement scales. RESULTS: The switch from sertraline to imipramine (mean dosage, 221 mg/d) and from imipramine to sertraline (mean dosage, 163 mg/d) resulted in clinically and statistically significant improvements. The switch to sertraline treatment was associated with fewer adverse effect complaints and significantly less attrition owing to adverse effects. Although sertraline treatment also resulted in significantly higher response rates in the intent-to-treat samples (60% in the sertraline group and 44% in the imipramine group), neither the intent-to-treat remission rates nor the response and remission rates among study completers differed significantly. Moreover, after considering the effect of attrition, there were no significant treatment effects on the more comprehensive generalized estimating equation analyses of the continuous dependent measures. CONCLUSIONS: More than 50% of chronically depressed antidepressant nonresponders benefited from a switch from imipramine to sertraline, or vice versa, despite a high degree of chronicity. As in the initial trial, sertraline was generally better tolerated than imipramine. Switching to a standard antidepressant of a different class is a useful treatment strategy for antidepressant nonresponders and could be considered a standard of comparison for future studies of novel alternate strategies. PMID- 11879162 TI - The etiology of phobias: an evaluation of the stress-diathesis model. AB - BACKGROUND: We evaluated for phobias the prediction of the stress-diathesis model that the magnitude of stress at onset is inversely proportional to the level of underlying diathesis. METHODS: In more than 7500 twins from a population-based registry, we assessed the personality trait of neuroticism--as an index of phobia proneness--and the lifetime histories of 5 phobia subtypes (agoraphobia, social, animal, situational, and blood or injury) and their associated irrational fears. Interviewers classified the mode of acquisition of the fear in phobic twins into 5 possible categories: trauma to self (further divided by severity), observed trauma to others, observed fear in others, taught by others to be afraid, and no memory of how or why fear developed. Analyses were conducted by logistic regression and analysis of covariance. RESULTS: The mode of acquisition had moderate test-retest reliability and differed meaningfully across phobia subtypes. None of the 3 tests of the stress-diathesis model was confirmatory: (1) the risk of phobias was not elevated in co-twins of twins who had no memory of their mode of acquisition, (2) the risk of phobias was not decreased in co-twins of twins who had severe trauma to self, and (3) no significant relationship, in phobic twins, was found between levels of neuroticism and mode of acquisition. CONCLUSIONS: These results are inconsistent with the traditional etiologic theories for phobias, which assume conditioning or social transmission. However, they are compatible with nonassociative models, which postulate that the vulnerability to phobias is largely innate and does not arise directly from environmental experiences. The stress-diathesis model may not be an appropriate paradigm for phobic disorders. PMID- 11879163 TI - Differential cerebral metabolic changes with paroxetine treatment of obsessive compulsive disorder vs major depression. AB - BACKGROUND: Serotonin reuptake inhibitors (SRIs) effectively treat both major depressive disorder (MDD) and obsessive-compulsive disorder (OCD). We compared and contrasted the functional neuroanatomical effects of SRIs in OCD and MDD as these 2 disorders occurred separately and concurrently by measuring pretreatment to posttreatment cerebral glucose metabolic changes in OCD vs MDD vs concurrent OCD + MDD. METHODS: We obtained [(18)F]fluorodeoxyglucose positron emission tomography (PET) brain scans on 25 subjects with OCD, 25 with MDD, and 16 with concurrent OCD + MDD before and after 8 to 12 weeks of treatment with paroxetine hydrochloride. Controls (n = 16) were scanned 10 to 12 weeks apart without treatment. Treatment response was defined as a more than 25% decline in OCD symptom severity, a more than 50% decline in MDD severity, and "much improved" clinical global impression. RESULTS: Although all patient groups received the same paroxetine dose for the same duration, regional metabolic changes differed significantly among diagnostic groups. Subjects with OCD alone showed significant metabolic decreases in the right caudate nucleus, right ventrolateral prefrontal cortex (VLPFC), bilateral orbitofrontal cortex, and thalamus that were not seen in any other group. Both the MDD and concurrent OCD + MDD groups showed metabolic decreases in the left VLPFC and increases in the right striatum. Treatment response was associated with a decrease in striatal metabolism in nondepressed OCD patients but with an increase in striatal activity in patients with OCD + MDD. CONCLUSIONS: Brain metabolic responses to SRIs are both disorder-specific and response-specific. They vary according to the underlying pathophysiology of the patient and the degree of symptomatic improvement. PMID- 11879165 TI - Improving clinical trials: American Society of Clinical Psychopharmacology recommendations. AB - The major purpose of this American Society of Clinical Psychopharmacology sponsored meeting was to identify strategies for more efficiently detecting clinical drug effects, thus reducing the economic and scientific risks of investigating new chemical entities in psychiatric disorders. The meeting consisted of presentations and discussions by experts who repeatedly had difficulty pursuing scientific, public health--relevant goals. Many approaches to improving the detection of potentially beneficial agents were reviewed. In this article, we discuss technically feasible study improvements. The scope of inquiry included identifying means of shifting institutional and regulatory assumptions and processes, even to the point of seeking appropriate national incentives. PMID- 11879164 TI - National Depressive and Manic-Depressive Association consensus statement on the use of placebo in clinical trials of mood disorders. AB - A consensus conference on the use of placebo in mood disorder studies consisted of expert presentations on bioethics, biostatistics, unipolar depression, and bipolar disorder. Work groups considered evidence and presented statements to the group. Although it was not possible to write a document for which there was complete agreement on all issues, the final document incorporated input from all authors. There was consensus that placebo has a definite role in mood disorder studies. Findings of equivalence between a new drug and standard treatment in active control studies is not evidence of efficacy unless the new drug is also significantly more effective than placebo. Add-on studies in which patients are randomized to standard therapy plus the investigational drug or standard therapy plus placebo are especially indicated for high-risk patients. Mood disorders in elderly and pediatric patients are understudied, and properly designed trials are urgently needed. Research is needed on the ethical conduct of studies to limit risks of medication-free intervals and facilitate poststudy treatment. Patients must fully understand the risks and lack of individualized treatment involved in research. PMID- 11879166 TI - Not in our methods, but in our ignorance. PMID- 11879167 TI - Out of the box. PMID- 11879168 TI - From clinical trial to prescription. AB - There are many steps between drug discovery and prescribing for a patient. Each step has problems. In this issue of ARCHIVES, Klein and colleagues propose changes in clinical trials that would result in more meaningful information for the treating physician. Of particular importance is the gap between what a physician needs to know and what is produced in the clinical trials that leads to approval of a new drug by the US Food and Drug Administration (FDA). Their recommendations for improving clinical trials are cogent, but broad-based implementation depends on an organizational structure and political effectiveness not presently in place. Most important, as the authors note, is an effective work group representing industry, regulatory agencies, and academic and federal science, addressing clinical trials issues with public participation. While improved quality and relevance of data from clinical trials will strengthen the scientific foundation of pharmacotherapy, other problems impede the delivery of objective information to the treating physician. Most hotly debated is the role of money in the creation and dissemination of knowledge. PMID- 11879169 TI - D-lactate and the false suffocation alarm. PMID- 11879171 TI - Prolonged brainstem auditory evoked potentials: an autism-specific or autism nonspecific marker. PMID- 11879173 TI - Apolipoprotein E epsilon4 allele and clinically defined vascular depression. PMID- 11879174 TI - Acute tryptophan depletion in schizophrenic patients treated with clozapine. PMID- 11879175 TI - Is psychosis exacerbated by modafinil? PMID- 11879176 TI - Burning charcoal: a novel and contagious method of suicide in Asia. PMID- 11879178 TI - A dual-specificity isoform of the protein kinase inhibitor PKI produced by alternate gene splicing. AB - We have previously shown that the protein kinase inhibitor beta (PKIbeta) form of the cAMP-dependent protein kinase inhibitor exists in multiple isoforms, some of which are specific inhibitors of the cAMP-dependent protein kinase, whereas others also inhibit the cGMP-dependent enzyme [Kumar, Van Patten and Walsh (1997), J. Biol. Chem. 272, 20011-20020]. We have now demonstrated that the switch from a cAMP-dependent protein kinase (PKA)-specific inhibitor to one with dual specificity arises as a consequence of alternate gene splicing. We have confirmed using bacterially produced pure protein that a single inhibitor species has dual specificity for both PKA and cGMP-dependent protein kinase (PKG), inhibiting each with very high and closely similar inhibitory potencies. The gene splicing converted a protein with 70 amino acids into one of 109 amino acids, and did not change the inhibitory potency to PKA, but changed it from a protein that had no detectable PKG inhibitory activity to one that now inhibited PKG in the nanomolar range. PMID- 11879179 TI - Crystal structures of 7-methylguanosine 5'-triphosphate (m(7)GTP)- and P(1)-7 methylguanosine-P(3)-adenosine-5',5'-triphosphate (m(7)GpppA)-bound human full length eukaryotic initiation factor 4E: biological importance of the C-terminal flexible region. AB - The crystal structures of the full-length human eukaryotic initiation factor (eIF) 4E complexed with two mRNA cap analogues [7-methylguanosine 5'-triphosphate (m(7)GTP) and P(1)-7-methylguanosine-P(3)-adenosine-5',5'-triphosphate (m(7)GpppA)] were determined at 2.0 A resolution (where 1 A=0.1 nm). The flexibility of the C-terminal loop region of eIF4E complexed with m(7)GTP was significantly reduced when complexed with m(7)GpppA, suggesting the importance of the second nucleotide in the mRNA cap structure for the biological function of eIF4E, especially the fixation and orientation of the C-terminal loop region, including the eIF4E phosphorylation residue. The present results provide the structural basis for the biological function of both N- and C-terminal mobile regions of eIF4E in translation initiation, especially the regulatory function through the switch-on/off of eIF4E-binding protein-eIF4E phosphorylation. PMID- 11879180 TI - Substrate access channel topology in membrane-bound prostacyclin synthase. AB - Results from our molecular-modelling and site-directed-mutagenesis studies of prostaglandin I(2) synthase (PGIS) have suggested that the large PGIS cytoplasmic domain is anchored to the endoplasmic reticulum (ER) membrane by the N-terminal segment in a way that orients the substrate access channel opening to face the membrane. To test this hypothesis we have explored the accessibility of the PGIS substrate channel opening to site-specific antibodies. The working three dimensional PGIS model constructed by protein homology modelling was used to predict surface portions near the substrate access channel opening. Two peptides corresponding to the surface immediately near the opening [residues 66-75 (P66 75) and 95-116 (P95-116)], and two other peptides corresponding to the surface about 10-20 A (1 A=0.1 nm) away from the opening [residues 366-382 (P366-382) and 472-482 (P472-482)] were used to prepare site-specific antibodies. All four antipeptide antibodies specifically recognized the synthetic segments of human PGIS and recombinant PGIS, as shown by binding assays and Western-blot analysis. The site-specific antibodies were used to probe the accessibility of the substrate access channel opening in transiently transfected COS-1 cells expressing recombinant human PGIS, and in spontaneously transformed human endothelial cell line ECV cells expressing endogenous human PGIS. Immunofluorescence staining was performed for cells selectively permeabilized with streptolysin O and for cells whose membranes were permeabilized with detergent. Antibodies to peptides in the immediate vicinity of the substrate channel (P66-75 and P95-116) bound to their targets only after general permeabilization with Triton X-100. In contrast, the two antibodies to peptides further from the channel opening (P366-382 and P472-482) bound to their targets even in cells with intact ER membranes. These observations support our topology model in which the PGIS substrate access channel opening is positioned close to the ER membrane. PMID- 11879181 TI - Essential stations in the intracellular pathway of cytotoxic bovine seminal ribonuclease. AB - Bovine seminal RNase (BS-RNase) is a dimeric RNase selectively cytotoxic for malignant cells. No information is available on its pathway from the extracellular matrix through the cytosol, where it degrades rRNA. An investigation of this pathway is reported here, carried out by immunofluorescence studies, by assessing the effects on BS-RNase cytotoxicity of drugs that affect specific intracellular compartments and by assaying the behaviour of a protein variant, BS-RNase-KDEL (BS-RNase in which a Lys-Asp-Glu-Leu peptide segment is inserted at the C-terminal ends of the subunit chains), endowed with a consensus sequence that directs proteins to the endoplasmic reticulum. BS-RNase was found to bind both normal and malignant cells and to be internalized by both cell types in endosome vesicles. Non-cytotoxic RNases, such as RNase A and a monomeric derivative of BS-RNase, did not bind to the cell surface and were not internalized. However, an engineered, dimeric and cytotoxic variant of RNase A bound effectively and permeated cells. The results of immunofluorescence studies, the effects of nigericin, monensin and brefeldin A on the cytotoxic action of seminal RNase, and the behaviour of the BS-RNase-KDEL variant, led to the conclusion that the pathway of BS-RNase in malignant cells from the extracellular matrix to the cytosol has two essential intracellular stations: endosomes and the trans-Golgi network. In normal cells, however, the protein does not progress from the endosomal compartment to the Golgi complex. PMID- 11879177 TI - The glucose-6-phosphatase system. AB - Glucose-6-phosphatase (G6Pase), an enzyme found mainly in the liver and the kidneys, plays the important role of providing glucose during starvation. Unlike most phosphatases acting on water-soluble compounds, it is a membrane-bound enzyme, being associated with the endoplasmic reticulum. In 1975, W. Arion and co workers proposed a model according to which G6Pase was thought to be a rather unspecific phosphatase, with its catalytic site oriented towards the lumen of the endoplasmic reticulum [Arion, Wallin, Lange and Ballas (1975) Mol. Cell. Biochem. 6, 75--83]. Substrate would be provided to this enzyme by a translocase that is specific for glucose 6-phosphate, thereby accounting for the specificity of the phosphatase for glucose 6-phosphate in intact microsomes. Distinct transporters would allow inorganic phosphate and glucose to leave the vesicles. At variance with this substrate-transport model, other models propose that conformational changes play an important role in the properties of G6Pase. The last 10 years have witnessed important progress in our knowledge of the glucose 6-phosphate hydrolysis system. The genes encoding G6Pase and the glucose 6-phosphate translocase have been cloned and shown to be mutated in glycogen storage disease type Ia and type Ib respectively. The gene encoding a G6Pase-related protein, expressed specifically in pancreatic islets, has also been cloned. Specific potent inhibitors of G6Pase and of the glucose 6-phosphate translocase have been synthesized or isolated from micro-organisms. These as well as other findings support the model initially proposed by Arion. Much progress has also been made with regard to the regulation of the expression of G6Pase by insulin, glucocorticoids, cAMP and glucose. PMID- 11879182 TI - Activation of c-Jun N-terminal kinase promotes survival of cardiac myocytes after oxidative stress. AB - Reperfusion injury occurs when ischaemic tissue is reperfused. It involves the generation and release of reactive oxygen that activates numerous signalling pathways and initiates cell death. Exposure of isolated cardiac myocytes to chronic hypoxia followed by reoxygenation results in the early activation of c Jun N-terminal kinase (JNK) and death by apoptosis of approx. 30% of the myocytes. Although JNK activation has been described in a number of models of ischaemia/reperfusion, the contribution of JNK activation to cell fate has not been established. Here we report that the activation of JNK by reoxygenation correlates with myocyte survival. Transfection of myocytes with JNK pathway interfering plasmid vectors or infection with adenoviral vectors support the hypothesis that JNK is protective. Transfection or infection with JNK inhibitory mutants increased the rates of apoptosis by almost 2-fold compared with control cultures grown aerobically or subjected to hypoxia and reoxygenation. Caspase 9 activity, measured by LEHD cleavage, increased >3-fold during reoxygenation and this activity was enhanced significantly at all times in cultures infected with dominant negative JNK adenovirus. Hypoxia-reoxygenation mediated a biphasic (2.6- and 2.9-fold) activation of p38 mitogen-activated protein kinase, as well as a small increase of tumour necrosis factor alpha (TNFalpha) secretion, but treatments with the p38 MAPK-specific inhibitor SB203580 or saturating levels of a TNFalpha-1 blocking antibody provided only partial protection against apoptosis. The results suggest that JNK activation is protective and that the pathway is largely independent of p38 MAPK or secreted TNFalpha. PMID- 11879183 TI - Bisphenol A diglycidyl ether induces apoptosis in tumour cells independently of peroxisome proliferator-activated receptor-gamma, in caspase-dependent and independent manners. AB - Peroxisome proliferator-activated receptors (PPARs) are nuclear transcription factors which are involved in many biological processes, such as regulation of cell differentiation, lipid metabolism, inflammation and cell death. PPARs consist of three families, PPAR-alpha, PPAR-delta and PPAR-gamma. Bisphenol A diglycidyl ether (BADGE) has been described as a pure antagonist of PPAR-gamma. However, recent data also revealed PPAR-gamma-agonistic activities of BADGE. Here we show that BADGE kills transformed cells by apoptosis and promotes the cytotoxic effects of tumour necrosis factor-related apoptosis-inducing ligand (TRAIL) and indomethacin. The cytotoxic effect of BADGE does not require PPAR gamma expression and is mediated in caspase-dependent and caspase-independent manners. PMID- 11879184 TI - Impaired chromosome partitioning and synchronization of DNA replication initiation in an insertional mutant in the Vibrio harveyi cgtA gene coding for a common GTP-binding protein. AB - The Vibrio harveyi cgtA gene product belongs to a subfamily of small GTP-binding proteins, called Obg-like proteins. Members of this subfamily are present in diverse organisms ranging from bacteria to humans. On the other hand, the functions of these proteins in the regulation of cellular processes are largely unknown. Genes coding for these proteins are essential in almost all bacteria investigated thus far. However, a viable V. harveyi insertional mutant in the cgtA gene was described recently. Therefore, this mutant gives a unique opportunity to study functions of a member of the subfamily of Obg-like proteins. Here we demonstrate that the mutant cells often form long filaments with expanded, non-partitioned or rarely partitioned chromosomes. Such a phenotype suggests impairment of the mechanism of chromosome partition. Flow cytometric studies revealed that synchronization of chromosome replication initiation is also significantly disturbed in the cgtA mutant. Moreover, in contrast to wild type V. harveyi, inhibition of chromosome replication and/or of cell division in the mutant bacteria caused significant increase in the number of large cells, suggesting that the cgtA gene product may be involved in the coupling of cell growth to chromosome replication and cell division. These results indicate that CgtA, an Obg-like GTP-binding protein, plays an important role in the regulation of chromosomal functions. PMID- 11879185 TI - Epitope-specific antibody-induced cleavage of angiotensin-converting enzyme from the cell surface. AB - Angiotensin I-converting enzyme (ACE; CD143, EC 3.4.15.1) is a type-1 integral membrane protein that can also be released into extracellular fluids (such as plasma, and seminal and cerebrospinal fluids) as a soluble enzyme following cleavage mediated by an unidentified protease(s), referred to as ACE secretase, in a process known as "shedding". The effects of monoclonal antibodies (mAbs) to eight different epitopes on the N-terminal domain of ACE on shedding was investigated using Chinese hamster ovary cells (CHO cells) expressing an ACE transgene and using human umbilical vein endothelial cells. Antibody-induced shedding of ACE was strongly epitope-specific: most of the antibodies increased the shedding by 20-40%, mAbs 9B9 and 3A5 increased the shedding by 270 and 410% respectively, whereas binding of mAb 3G8 decreased ACE shedding by 36%. The ACE released following mAb treatment lacked a hydrophobic transmembrane domain anchor. The antibody-induced shedding was completely inhibited at 4 degrees C and by zinc chelation using 1,10-phenanthroline, suggesting involvement of a metalloprotease in this process. A hydroxamate-based metalloprotease inhibitor (batimastat, BB-94) was 15 times more efficacious in inhibiting mAb-induced ACE shedding than basal (constitutive) ACE release. Treatment of CHO-ACE cells with BB-94 more effectively prevented elevation in antibody-dependent (but not basal) ACE release induced by 3,4-dichloroisocoumarin and iodoacetamide. These data suggest that different secretases might be responsible for ACE release under basal compared with antibody-induced shedding. Further experiments with more than 40 protease inhibitors suggest that calpains, furin and the proteasome may participate in this process. PMID- 11879186 TI - The beta-appendages of the four adaptor-protein (AP) complexes: structure and binding properties, and identification of sorting nexin 9 as an accessory protein to AP-2. AB - Adaptor protein (AP) complexes are essential components for the formation of coated vesicles and the recognition of cargo proteins for intracellular transport. Each AP complex exposes two appendage domains with that function to bind regulatory accessory proteins in the cytosol. Secondary structure predictions, sequence alignments and CD spectroscopy were used to relate the beta appendages of all human AP complexes to the previously published crystal structure of AP-2. The results suggested that the beta-appendages of AP-1, AP-2 and AP-3 have similar structures, consisting of two subdomains, whereas that of AP-4 lacks the inner subdomain. Pull-down and overlay assays showed partial overlap in the binding specificities of the beta-appendages of AP-1 and AP-2, whereas the corresponding domain of AP-3 displayed a unique binding pattern. That AP-4 may have a truncated, non-functional domain was indicated by its apparent inability to bind any proteins from cytosol. Of several novel beta-appendage binding proteins detected, one that had affinity exclusively for AP-2 was identified as sorting nexin 9 (SNX9). SNX9, which contains a phox and an Src homology 3 domain, was found in large complexes and was at least partially associated with AP-2 in the cytosol. SNX9 may function to assist AP-2 in its role at the plasma membrane. PMID- 11879187 TI - A region in the first exon/intron of rat carnitine palmitoyltransferase Ibeta is involved in enhancement of basal transcription. AB - Carnitine palmitoyltransferase-Ibeta (CPT-Ibeta) catalyses the transfer of long chain fatty acids to the enzymes of beta-oxidation of muscle and heart. Transcriptional control of this regulatory protein is relevant to disorders of fatty acid oxidation and the switch to glucose metabolism that occurs in cardiac pathology. The presence of a transcriptional enhancer sequence in the first untranslated exon and first intron of the CPT-Ibeta gene was identified using deletional and mutational analysis, and by ligation of an oleate responsive element (fatty acid response element) to a minimal promoter. The enhancer sequences are contained in the first 40 bases downstream of the transcription start site and increase CPT-Ibeta reporter gene expression independent of any 5' cis-acting elements. Deletion of the first 40 bases of the 3'-untranslated region does not affect the up-regulation of transcription by 10 microM phenylephrine. However, mutation and/or deletion of bases between +11 and +30 dramatically decreases reporter gene expression. Electrophoretic mobility-shift assays reveal two DNA (+11 to +36)-protein complexes that appear cardiac specific. The exon/intron element enhances activation of the heterologous thymidine kinase promoter in a position- and orientation-dependent manner. Therefore we have identified a novel region in the first exon/intron of the CPT-Ibeta gene that acts as a non-classical transcriptional enhancer downstream of regulatory elements characterized previously in the 5'-flanking region of the minimal promoter. PMID- 11879189 TI - Macrophage metalloproteinases degrade high-density-lipoprotein-associated apolipoprotein A-I at both the N- and C-termini. AB - Atheromatous plaques contain various cell types, including macrophages, endothelial cells and smooth-muscle cells. To investigate the possible interactions between secreted matrix metalloproteinases and high-density lipoprotein (HDL) components, we tested the above cell types by culturing them for 24 h. HDL(3) (HDL subfractions with average sizes of between 8.44 nm for HDL(3A) and 7.62 nm for HDL(3C)) were then incubated in their cell-free conditioned media. Proteolytic degradation of apolipoprotein A-I was observed with macrophages, but not with endothelial-cell- or muscle-cell-conditioned supernatant. Absence of calcium or addition of EDTA to incubation media prevented all proteolytic processes. The identified apolipoprotein A-I fragments had sizes of 26, 22, 14 and 9 kDa. Two-dimensional electrophoresis and MS resolved the 26 and the 22 kDa components and identified peptides resulting from both N- and C terminal cleavage of apolipoprotein A-I. The higher abundance of C- than N terminally cleaved peptides agrees with data in the literature for a fully structured alpha-helix around Tyr(18) compared with an unstructured region around Gly(185) and Gly(186). The flexibility in the latter region of apolipoprotein A-I may explain its susceptibility to proteolysis. In our experimental set-up, HDL(3C) was more extensively degraded than the other HDL(3) subclasses (HDL(3A) and HDL(3B)). Proteolytic fragments produced by metalloproteinase action were shown by gel filtration and electrophoresis to be neither associated with lipids nor self-associated. PMID- 11879188 TI - Activation of bacterial ceramidase by anionic glycerophospholipids: possible involvement in ceramide hydrolysis on atopic skin by Pseudomonas ceramidase. AB - We have reported previously that the ceramidase from Pseudomonas aeruginosa AN17 isolated from a patient with atopic dermatitis requires detergents for hydrolysis of ceramide (Cer) [Okino, Tani, Imayama and Ito (1998) J. Biol. Chem. 273, 14368- 14373]. In the present study, we report that some glycerophospholipids strongly activated the hydrolysis of Cer by Pseudomonas ceramidase in the absence of detergents. Among the glycerophospholipids tested, cardiolipin was most effective in stimulating hydrolysis of Cer followed by phosphatidic acid, phosphatidylethanolamine and phosphatidylglycerol, whereas phosphatidylcholine, lysophosphatidic acid and diacylglycerol were less effective. Interestingly, Staphylococcus aureus-derived lipids, which contain cardiolipin and phosphatidylglycerol as major lipid components, also strongly enhanced the hydrolysis of normal Cer, as well as the human skin-specific omega-hydroxyacyl Cer, by the enzyme in the absence of detergents. It was confirmed that several strains of P. aeruginosa, including AN17, secrete a significant amount of staphylolytic proteases to lyse S. aureus cells, resulting in the release of cardiolipin and phosphatidylglycerol. Since both P. aeruginosa and S. aureus are suspected of being present in microflora of atopic skin, we speculate that S. aureus-derived glycerophospholipids stimulate the hydrolysis of Cer in atopic skin by bacterial ceramidase. PMID- 11879190 TI - Heat-shock protein 70 attenuates nitric oxide-induced apoptosis in RAW macrophages by preventing cytochrome c release. AB - Heat-shock protein (Hsp) 70 is an inhibitor of apoptosis and has been shown to protect against nitric oxide-mediated toxicity. To gain mechanistic insights into the actions of Hsp70, we stably transfected RAW 264.7 mouse macrophages with the human Hsp70 gene and investigated critical steps in the progression towards cell demise. Incubation of control and Hsp70-transfected macrophages with S nitrosoglutathione induced accumulation of the tumour suppressor p53, expression of p21(WAF1/CIP1) (where WAF1 corresponds to wild-type p53-activated fragment 1 and CIP1 corresponds to cyclin-dependent kinase-interacting protein 1) and G(1) cell-cycle arrest. However, cytochrome c translocation to the cytosol and activation of caspase 9 and caspase 3 were markedly reduced in Hsp70 overexpressing cells. In addition, changes in nuclear morphology, as determined by Hoechst staining, and the appearance of cells in the sub-G(1) phase were diminished in Hsp70-overexpressing cells compared with controls. We conclude that, in macrophages, Hsp70 interferes with cytochrome c release from mitochondria and, thereby, prevents nitric oxide-induced apoptosis, but leaves p53 accumulation and interference in the cell cycle intact. PMID- 11879192 TI - GTP binds to Rab3A in a complex with Ca2+/calmodulin. AB - Ras-like small GTP-binding proteins of the Rab family regulate trafficking of the secretory or endocytic pathways. Rab3 proteins within the Rab family are expressed at high levels in neurons and endocrine cells, where they regulate release of dense-core granules and synaptic vesicles (SVs). Rab3A is present as either the soluble or the SV membrane-bound form in neurons that are dependent on the GDP- or GTP-bound states respectively. GDP dissociation inhibitor (GDI) is known to induce the dissociation of Rab3A from synaptic membranes when GTP is depleted. In an earlier study, Ca(2+)/calmodulin (CaM) was also shown to dissociate Rab3A from synaptic membranes by forming an equimolar complex with Rab3A in vitro. We have examined a possible role for Ca(2+)/CaM in modulating both the binding of guanine nucleotides to Rab3A and the GTPase activity of Rab3A. The basal level of Rab3A GTPase activity was not affected by an association with Ca(2+)/CaM. Ca(2+)/CaM-Rab3A complex that was formed in synaptic membranes was able to bind guanine nucleotides, whereas the Rab3A-GDI complex could not. In addition, Ca(2+)/CaM led to the replacement of the GDP molecule in the Rab3A-GDI complex with GTP in Rab3A. Taken together, these results suggest that CaM may have a role in stimulating GTP binding to Rab3A that is complexed with GDI, which leads to the formation of an active GTP-bound form of the Rab3A Ca(2+)/CaM complex. PMID- 11879191 TI - Decorin suppresses transforming growth factor-beta-induced expression of plasminogen activator inhibitor-1 in human mesangial cells through a mechanism that involves Ca2+-dependent phosphorylation of Smad2 at serine-240. AB - Transforming growth factor-beta (TGFbeta) is a key mediator of extracellular matrix (ECM) accumulation in sclerotic kidney diseases such as diabetic nephropathy. One of the main target cells for TGFbeta in the kidney are glomerular mesangial cells, which respond by increasing expression of ECM proteins, such as collagens, laminin and fibronectin, while suppressing the expression of ECM-degrading proteases and increasing the synthesis of ECM protease inhibitors, including plasminogen activator inhibitor-1. Previous studies have shown that exposure of mesangial cells to chronic high-glucose conditions, such as those seen in diabetes, increases ECM deposition in a mechanism involving glucose-mediated up-regulation of TGFbeta expression. Naturally occurring inhibitors of this TGFbeta-dependent fibrotic response include decorin, a small leucine-rich proteoglycan. While the mechanism by which TGFbeta stimulates gene expression via the Smad signal-transduction pathway is becoming clear, the precise mechanism by which decorin may impinge upon TGFbeta activity remains to be established. In this study, for the first time we provide evidence that decorin can disrupt glucose- and TGFbeta/Smad-dependent transcriptional events in human mesangial cells through a mechanism that involves an increase in Ca(2+) signalling, the activation of Ca(2+)/calmodulin-dependent protein kinase II and ensuing phosphorylation of Smad2 at Ser-240. We show that decorin also induces Ser-240 phospho-Smad hetero-oligomerization with Smad4 and the nuclear localization of this complex independently of TGFbeta receptor activation. Thus, in human mesangial cells, the mechanism of decorin-mediated inhibition of TGFbeta signalling may involve activation of Ca(2+) signalling, the subsequent phosphorylation of Smad2 at a key regulatory site, and the sequestration of Smad4 in the nucleus. PMID- 11879193 TI - Cinnamomin, a type II ribosome-inactivating protein, is a storage protein in the seed of the camphor tree (Cinnamomum camphora). AB - Cinnamomin is a novel type II ribosome-inactivating protein (RIP) isolated in our laboratory from the seed of the camphor tree (Cinnamomum camphora). In this paper the physiological role it plays in the plant cell was studied. Northern and Western blotting revealed that cinnamomin was expressed specifically in cotyledons. It accumulated in large amounts simultaneously with other proteins at the post-stages of seed development. Cinnamomin degraded rapidly during the early stages of seed germination. Endopeptidase was proved to play an important role in the degradation of cinnamomin. Western blotting of total proteins from the protein body with antibodies against cinnamomin demonstrated that it only existed in this specific cellular organelle as a storage protein. The similar properties of cinnamomin and other seed storage proteins of dicotyledons were compared. We conclude that cinnamomin is a special storage protein in the seed of C. camphora. PMID- 11879194 TI - Sorbitol activates atypical protein kinase C and GLUT4 glucose transporter translocation/glucose transport through proline-rich tyrosine kinase-2, the extracellular signal-regulated kinase pathway and phospholipase D. AB - Sorbitol, "osmotic stress", stimulates GLUT4 glucose transporter translocation to the plasma membrane and glucose transport by a phosphatidylinositol (PI) 3-kinase independent mechanism that reportedly involves non-receptor proline-rich tyrosine kinase-2 (PYK2) but subsequent events are obscure. In the present study, we found that extracellular signal-regulated kinase (ERK) pathway components, growth factor-receptor-bound-2 protein, son of sevenless (SOS), RAS, RAF and mitogen activated protein (MAP) kinase/ERK kinase, MEK(-1), operating downstream of PYK2, were required for sorbitol-stimulated GLUT4 translocation/glucose transport in rat adipocytes, L6 myotubes and 3T3/L1 adipocytes. Furthermore, sorbitol activated atypical protein kinase C (aPKC) through a similar mechanism depending on the PYK2/ERK pathway, independent of PI 3-kinase and its downstream effector, 3-phosphoinositide-dependent protein kinase-1 (PDK-1). Like PYK2/ERK pathway components, aPKCs were required for sorbitol-stimulated GLUT4 translocation/glucose transport. Interestingly, sorbitol stimulated increases in phospholipase D (PLD) activity and generation of phosphatidic acid (PA), which directly activated aPKCs. As with aPKCs and glucose transport, sorbitol stimulated PLD activity was dependent on the ERK pathway. Moreover, PLD-generated PA was required for sorbitol-induced activation of aPKCs and GLUT4 translocation/glucose transport. Our findings suggest that sorbitol sequentially activates PYK2, the ERK pathway and PLD, thereby increasing PA, which activates aPKCs and GLUT4 translocation. This mechanism contrasts with that of insulin, which primarily uses PI 3-kinase, D3-PO(4) polyphosphoinositides and PDK-1 to activate aPKCs. PMID- 11879195 TI - A novel function for the 90 kDa heat-shock protein (Hsp90): facilitating nuclear export of 60 S ribosomal subunits. AB - Ribosomal subunits are assembled in the nucleus, and mature 40 S and 60 S subunits are exported stoichiometrically into the cytoplasm. The nuclear export of ribosomal subunits is a unidirectional, saturable and energy-dependent process. An in vitro assay for the nuclear export of 60 S ribosomal subunits involves the use of resealed nuclear envelopes. The export of ribosomal subunits from resealed nuclear envelopes is enhanced by cytoplasmic proteins. Here we present evidence that the export-promoting activity was due to the cytoplasmic 90 kDa heat-shock protein (Hsp90). Isolated, purified Hsp90 vastly enhanced the export of 60 S ribosomal subunits from resealed nuclear envelopes, while inhibition of Hsp90 function, either with the Hsp90-binding drug geldanamycin or with anti-Hsp90 antibodies, resulted in reduced release of 60 S ribosomal subunits. To confirm these findings under in vivo conditions, corresponding experiments were performed with Xenopus oocytes using microinjection techniques; the results obtained confirmed the findings obtained with resealed nuclear envelopes. These findings suggest that Hsp90 facilitates the nuclear export of 60 S ribosomal subunits, probably by chaperoning protein interactions during the export process. PMID- 11879196 TI - Mode of inhibition of HIV-1 reverse transcriptase by polyacetylenetriol, a novel inhibitor of RNA- and DNA-directed DNA polymerases. AB - Polyacetylenetriol (PAT), a natural marine product from the Mediterranean sea sponge Petrosia sp., was found to be a novel general potent inhibitor of DNA polymerases. It inhibits equally well the RNA- and DNA-dependent DNA polymerase activities of retroviral reverse transcriptases (RTs) (i.e. of HIV, murine leukaemia virus and mouse mammary tumour virus) as well as cellular DNA polymerases (i.e. DNA polymerases alpha and beta and Escherichia coli polymerase I). A study of the mode and mechanism of the polymerase inhibition by PAT has been conducted with HIV-1 RT. PAT was shown to be a reversible non-competitive inhibitor. PAT binds RT independently and at a site different from that of the primer-template and dNTP substrates with high affinity (K(i)=0.51 microM and K(i)=0.53 microM with dTTP and with dGTP as the variable substrates respectively). Blocking the polar hydroxy groups of PAT has only a marginal effect on the inhibitory capacity, thus hydrophobic interactions are likely to play a major role in inhibiting RT. Preincubation of RT with the primer-template substrate prior to the interaction with PAT reduces substantially the inhibition capacity, probably by preventing these contacts. PAT does not interfere with the first step of polymerization, the binding of RT to DNA, nor does the inhibitor interfere with the binding of dNTP to RT/DNA complex, as evident from the steady state kinetic study, whereby K(m) remains unchanged. We assume, therefore, that PAT interferes with subsequent catalytic steps of DNA polymerization. The inhibitor may alter the optimal stereochemistry of the polymerase active site relative to the primer terminus, bound dNTP and the metal ions that are crucial for efficient catalysis or, alternatively, may interfere with the thumb sub domain movement and, thus, with the translocation of the primer-template following nucleotide incorporation. PMID- 11879197 TI - Mesangial cell gelatinase A synthesis is attenuated by oscillating hyperbaric pressure. AB - Glomerular hypertension has been established as a major factor contributing to glomerular scarring. Underlying cellular mechanisms leading to matrix accumulation are largely unknown. The isolated effect of oscillating hyperbaric pressure [OP; P(max) 50 mmHg (1 mmHg=0.133 kPa), P(mean) 24 mmHg, with a fixed oscillation of 60/min] on matrix-degrading protease secretion by rat mesangial cells (MCs) was analysed using a pressure chamber model described previously [Mertens, Espenkott, Venjakob, Heintz, Handt and Sieberth (1998) Hypertension 32, 945-952]. MCs were grown under atmospheric pressure (AP) or a controlled OP, and protease synthesis and gene transcription were analysed. A distinct biphasic cellular response to OP with stimulated gelatinase A protein expression and enzyme activity during the initial 24 h, and subsequent inhibition, was apparent, as shown by gelatin zymography. Gelatinase B activity remained unchanged. The abundance of gelatinase A transcripts, determined by reverse transcriptase-PCR, indicated a concordant regulation of gene transcription. To elucidate underlying regulatory events, reporter constructs were transfected. In these experiments, a recently identified response element, RE-1, conferred a significant stimulatory effect within the initial 4 h of OP. Nuclear protein/RE-1 binding studies revealed additional complexes from 5 min up to 3 h after OP exposure, with intensities dependent on P(max). STAT3 was identified as a component of these novel complexes. Down-regulation of cis-activity after 48 h of OP exposure was not transferred via the proximal 1686 bp of the gelatinase A regulatory sequence. In conclusion, hyperbaric OP elicits time-dependent changes in rat MC gelatinase A gene transcription. PMID- 11879198 TI - Evidence that cytosolic calcium increases are not sufficient to stimulate phospholipid scrambling in human T-lymphocytes. AB - Phospholipid scrambling, the disruption of normal plasma-membrane asymmetry, occurs during apoptotic and necrotic cell death and during the activation of platelets and neutrophils. It is currently believed that phospholipid scrambling is triggered simply by increases in bulk cytosolic [Ca(2+)]. We have presented evidence previously that the styryl dye FM1-43 is sensitive to phospholipid scrambling in Jurkat human leukaemic T-lymphocytes. Here we have used FM1-43, in combination with fura 2 and the Ca(2+)-elevating agents ionomycin and thapsigargin, in imaging experiments to test the idea that increases in bulk cytosolic [Ca(2+)] stimulate scrambling. Intracellular Ca(2+) increases of approximately 2 microM accompanied ionomycin-stimulated scrambling in approximately 50% of cells, and scrambling occurred in >99% of cells in which intracellular Ca(2+) rose to 4 microM. Chelating intracellular Ca(2+) with bis-(o aminophenoxy)ethane-N,N,N',N'-tetra-acetic acid or EGTA suppressed both ionomycin stimulated intra cellular Ca(2+) increases and scrambling, demonstrating that intracellular Ca(2+) increases are necessary for ionomycin-stimulated scrambling. However, elevating intracellular Ca(2+) to 2-4 microM with thapsigargin, a drug that depletes intracellular Ca(2+) stores and triggers Ca(2+) entry via Ca(2+) release-activated Ca(2+) channels, did not trigger scrambling, as assessed with either FM1-43 or FITC-labelled annexin V. These results suggest that increases in intracellular [Ca(2+)] are necessary but not sufficient to stimulate scrambling in lymphoyctes, and indicate that ionomycin has an additional effect that is required to stimulate scrambling. PMID- 11879200 TI - Mono-ADP-ribosyltransferases in human monocytes: regulation by lipopolysaccharide. AB - ADP-ribosyltransferase activity was shown to be present on the surface of human monocytes. Incubating the cells in the presence of BSA leads to an increase in enzyme activity. The acceptor amino acid mainly responsible for the ADP-ribose bond was identified as a cysteine residue. An increase in ADP-ribosyltransferase activity was observed when cells were treated for 16 h with bacterial lipopolysaccharide (LPS). Possible candidates for catalysing the reaction are mono-ADP-ribosyltransferases (ARTs). When measuring expression of the mRNA of ART1, 3, 4 and 5, only ART3 mRNA was detected in unstimulated monocytes. Upon stimulation for 16 h with LPS, lipoteichoic acid or peptidoglycan, ART4 mRNA was found to be expressed. No ART4 signal appeared after a 4 h exposure of the cells to LPS. Cell-surface proteins were labelled when incubating monocytes with [(32)P]NAD(+). Their molecular masses were 29, 33, 43, 45, 60 and 82 kDa. In response to LPS an additional protein of 31 kDa was found to be labelled. The bound label was resistant to treatment with NH(2)OH but sensitive to HgCl(2), characteristic of a cysteine-linked ADP-ribosylation. PMID- 11879199 TI - Alpha-tocopheryl succinate, an agent with in vivo anti-tumour activity, induces apoptosis by causing lysosomal instability. AB - Certain vitamin E analogues, such as alpha-tocopheryl succinate (alpha-TOS), exhibit in vivo anti-tumour activity and, in vitro, induce apoptosis of cultured tumour cells. In the present study we report that these effects may be explained, at least in part, by destabilization of lysosomal membranes. alpha-TOS, but not alpha-tocopheryl acetate or alpha-tocopherol (alpha-TOH), induced early lysosomal destabilization followed by apoptosis. Similar effects were observed with beta TOS, whereas beta-TOH was inactive. Cathepsin D-deficient cells were more resistant to alpha-TOS than their normal counterparts, and featured delayed caspase activation. Possible detergent and lysosomotropic effects of alpha- and beta-TOS were suggested by their haemolytic activity in an in vitro test and their release of beta-galactosidase from isolated lysosomes, whereas the non succinylated analogues were inactive. The pro-apoptotic activity of alpha-TOS was pH-dependent, being greater at lower pH, typical of the interstitium of solid tumours. These findings indicate that lysosomal destabilization may partially or fully explain the induction of apoptosis in cultured cells by alpha-TOS and the mechanism whereby this agent exerts in vivo anti-tumour effects. PMID- 11879201 TI - Mechanism of the regulation of type IB phosphoinositide 3OH-kinase byG-protein betagamma subunits. AB - Type IB phosphoinositide 3OH-kinase (PI3K) is activated by G-protein betagamma subunits (Gbetagammas). The enzyme is soluble and largely cytosolic in vivo. Its substrate, PtdIns(4,5)P(2), and the Gbetagammas are localized at the plasma membrane. We have addressed the mechanism by which Gbetagammas regulate the PI3K using an in vitro approach. We used sedimentation assays and surface plasmon resonance to determine association of type IB PI3K with lipid monolayers and vesicles of varying compositions, some of which had Gbetagammas incorporated. Association and dissociation rate constants were determined. Our results indicated that in an assay situation in vitro the majority of PI3K will be associated with lipid vesicles, irrespective of the presence or absence of Gbetagammas. In line with this, a constitutively active membrane-targeted PI3K construct could still be activated substantially by Gbetagammas in vitro. We conclude that Gbetagammas activate type IB PI3K by a mechanism other than translocation to the plasma membrane. PMID- 11879202 TI - The ratio between tetrahydrobiopterin and oxidized tetrahydrobiopterin analogues controls superoxide release from endothelial nitric oxide synthase: an EPR spin trapping study. AB - Augmentation of superoxide levels has been linked to impaired relaxation in hypertension, diabetes and hypercholesterolaemia. Purified endothelial nitric oxide synthase (eNOS) generates superoxide under limited availability of 5,6,7,8 tetrahydrobiopterin (BH(4)). Thus alterations in endothelial BH(4) levels have been postulated to stimulate superoxide production from eNOS. This possibility was examined by determining the concentration-dependent effects of BH(4), and its analogues, on superoxide formation by eNOS. Superoxide was quantified by EPR spin trapping, which is the only available technique to quantify superoxide from eNOS. Using 5-ethoxycarbonyl-5-methyl-pyrroline N-oxide, we show that only fully reduced BH(4) diminished superoxide release from eNOS, with efficiency BH(4)>6 methyl-BH(4)>5-methyl-BH(4). In contrast, partially oxidized BH(4) analogues, 7,8 dihydrobiopterin (7,8-BH(2)) and sepiapterin had no effect. Neither l-arginine nor N(G)-nitro-l-arginine methyl ester (l-NAME) abolished superoxide formation. Together, BH(4) and l-arginine stimulated .NO production at maximal rates of 148 nmol/min per mg of protein. These results indicate that BH(4) acts as a "redox switch", decreasing superoxide release and enhancing .NO formation. This role was verified by adding 7,8-BH(2) or sepiapterin to fully active eNOS. Both 7,8-BH(2) and sepiapterin enhanced superoxide release while inhibiting (.)NO formation. Collectively, these results indicate that the ratio between oxidized and reduced BH(4) metabolites tightly regulates superoxide formation from eNOS. The pathological significance of this scenario is discussed. PMID- 11879203 TI - Comparison between Ca2+-induced scrambling of various fluorescently labelled lipid analogues in red blood cells. AB - Treatment of red blood cells with calcium and ionomycin causes activation of the lipid scramblase, a putative membrane protein catalysing flip-flop of (phospho)lipids. Various fluorescent 1-oleoyl-2-[6(7-nitrobenz-2-oxa-1,3-diazol-4 yl)amino] caproyl (C(6)-NBD) analogues were tested for transbilayer movement across the plasma membrane of red blood cells. Among these phospholipid analogues were phosphatidylgalactose, phosphatidylmaltose and phosphatidylmaltotriose, which were obtained from C(6)-NBD-phosphatidylcholine by phospholipase D catalysed transphosphatidylation. The inward movement after the onset of scrambling was monitored by extraction of the non-internalized probe with BSA. We demonstrate that both the amino group and the size of the headgroup determine the kinetics of lipid scrambling, and that lipids with a ceramide backbone migrate much more slowly than glycerophospholipids with the same headgroup. PMID- 11879204 TI - A cytochrome c mutant with high electron transfer and antioxidant activities but devoid of apoptogenic effect. AB - A cytochrome c mutant lacking apoptogenic function but competent in electron transfer and antioxidant activities has been constructed. To this end, mutant species of horse and yeast cytochromes c with substitutions in the N-terminal alpha-helix or position 72 were obtained. It was found that yeast cytochrome c was much less effective than the horse protein in activating respiration of rat liver mitoplasts deficient in endogenous cytochrome c as well as in inhibition of H(2)O(2) production by the initial segment of the respiratory chain of intact rat heart mitochondria. The major role in the difference between the horse and yeast proteins was shown to be played by the amino acid residue in position 4 (glutamate in horse, and lysine in yeast; horse protein numbering). A mutant of the yeast cytochrome c containing K4E and some other "horse" modifications in the N-terminal alpha-helix, proved to be (i) much more active in electron transfer and antioxidant activity than the wild-type yeast cytochrome c and (ii), like the yeast cytochrome c, inactive in caspase stimulation, even if added in 400-fold excess compared with the horse protein. Thus this mutant seems to be a good candidate for knock-in studies of the role of cytochrome c-mediated apoptosis, in contrast with the horse K72R, K72G, K72L and K72A mutant cytochromes that at low concentrations were less active in apoptosis than the wild-type, but were quite active when the concentrations were increased by a factor of 2-12. PMID- 11879205 TI - Complete beta-oxidation of valproate: cleavage of 3-oxovalproyl-CoA by a mitochondrial 3-oxoacyl-CoA thiolase. AB - The beta-oxidation of valproic acid (VPA; 2-n-propylpentanoic acid) was investigated in vitro in intact rat liver mitochondria incubated with (3)H labelled VPA. The metabolism of [4,5-(3)H(2)]VPA and [2-(3)H]VPA was studied by analysing the different acyl-CoA intermediates formed by reverse-phase HPLC with radiochemical detection. Valproyl-CoA, Delta(2(E))-valproyl-CoA,3-hydroxyvalproyl CoA and 3-oxovalproyl-CoA (labelled and non-labelled) were determined using continuous on-line radiochemical and UV detection. The formation of these intermediates was investigated using the two tritiated precursors in respiratory states 3 and 4. Valproyl-CoA was present at highest concentrations under both conditions. Two distinct labelled peaks were found, which were identified as (3)H(2)O and [4,5-(3)H(2)]3-oxo-VPA. The formation of (3)H(2)O strongly suggested that VPA underwent complete beta-oxidation and that [4,5-(3)H(2)]3-oxo-VPA was formed by hydrolysis of the corresponding thioester. The hypothesis that 3 oxovalproyl-CoA undergoes thiolytic cleavage was investigated further. For this purpose a mito chondrial lysate was incubated with synthetic 3-oxovalproyl-CoA, carnitine and carnitine acetyltransferase for subsequent monitoring of the formation of propionylcarnitine and pentanoylcarnitine using electrospray ionization tandem MS. The detection of these compounds demonstrated unequivocally that the intermediate 3-oxovalproyl-CoA is a substrate of a mitochondrial thiolase, producing propionyl-CoA and pentanoyl-CoA, thus demonstrating the complete beta-oxidation of VPA in the mitochondrion. Our data should lead to a re evaluation of the generally accepted concept that the biotransformation of VPA by mitochondrial beta-oxidation is incomplete. PMID- 11879207 TI - When they got there the cupboard was bare. PMID- 11879206 TI - Further characterization of the interaction between the cytoskeletal proteins talin and vinculin. AB - The cytoskeletal protein talin, which is thought to couple integrins to F-actin, contains three binding sites (VBS1-VBS3) for vinculin, a protein implicated in the negative regulation of cell motility and whose activity is modulated by an intramolecular interaction between the vinculin head (Vh) and vinculin tail (Vt) domains. In the present study we show that recombinant talin polypeptides containing the three VBSs (VBS1, residues 498-636; VBS2, residues 727-965; and VBS3, residues 1943-2157) each bind tightly to the same or overlapping sites within vinculin(1-258). A short synthetic talin VBS3 peptide (residues 1944-1969) was sufficient to inhibit binding of a (125)I-labelled talin VBS3 polypeptide to vinculin(1-258), and NMR spectroscopy confirmed that this peptide forms a 1:1 complex in slow exchange with vinculin(1-258). Binding of the (125)I-labelled VBS3 polypeptide was markedly temperature dependent, but was not inhibited by 1 M salt or 10% (v/v) 2-methyl-2-propanol. Attempts to further define the talin binding site within vinculin(1-258) using a gel-blot assay were unsuccessful, but near maximal talin-binding activity was retained by a construct spanning vinculin residues 1-131 in a yeast two-hybrid assay. Interestingly, the talin VBS3 polypeptide was a potent inhibitor of the Vh-Vt interaction, and the VBS3 synthetic peptide was able to expose the actin-binding site in intact vinculin, which is otherwise masked by the Vh-Vt interaction. The results suggest that under certain conditions, talin may be an effective activator of vinculin. PMID- 11879208 TI - Remifentanil-sevoflurane anaesthesia for laparoscopic cholecystectomy: comparison of three dose regimens. AB - The objective of this study was to determine a dosing regimen for remifentanil sevoflurane anaesthesia that achieves an optimal balance between quality of anaesthesia and time to recovery. Patients undergoing elective laparoscopic cholecystectomy were randomly allocated to receive 0.4, 0.8 or 1.2 MAC (minimal alveolar concentration) of sevoflurane combined with remifentanil as required to maintain stable anaesthesia. For induction of anaesthesia, the remifentanil dose was 25 microg x kg(-1) x h(-1) and the mean propofol dose which was required to obtain loss of consciousness was 1.59 mg x kg(-1). During the maintenance phase, the mean remifentanil dose was 16.0, 14.1 and 13.0 microg x kg(-1) x h(-1) for the 0.4, 0.8 and 1.2 MAC groups, respectively. The mean sevoflurane maintenance dose was 0.91, 1.24 and 2.1% end-tidal for the 0.4, 0.8 and 1.2 MAC groups, respectively. The incidence of somatic responses was significantly higher in the 0.4 MAC sevoflurane group. Recovery times were significantly faster in the 0.4 compared to the 0.8 and 1.2 MAC groups and in the 0.8 compared to the 1.2 MAC group. The combination of 14 microg x kg(-1) x h(-1) remifentanil and 1.24% end tidal sevoflurane achieved the optimal balance between the quality, and recovery from anaesthesia. PMID- 11879209 TI - The effect of ulinastatin pre-treatment on vecuronium-induced neuromuscular block in patients with hepatic cirrhosis. AB - The aim of this study was to investigate the effect of ulinastatin, a protease inhibitor, on the neuromuscular block produced by vecuronium in patients with hepatic cirrhosis. Thirty adult patients with hepatic cirrhosis were randomly allocated to receive ulinastatin (cirrhosis/ulinastatin group, n = 15) or saline (cirrhosis/saline group, n = 15). Fifteen healthy adult patients without hepatic cirrhosis comprised a control group. Patients were given a standardised anaesthetic that included nitrous oxide and isoflurane in oxygen, and fentanyl. A bolus dose of ulinastatin 5000 unit x kg(-1) was given to members of the cirrhosis/ulinastatin group. The same volume of normal saline was given to the other two groups. Two minutes later, vecuronium 0.1 mg x kg(-1) was given. The onset of neuromuscular block was significantly slower in the cirrhosis/ulinastatin group than in the cirrhosis/saline and control groups (p < 0.05). Spontaneous recovery of neuromuscular function was significantly quicker in the cirrhosis/ulinastatin and control groups than in the cirrhosis/saline group (p < 0.05). The time course of recovery in the cirrhosis/ulinastatin and control groups was similar. We conclude that in cirrhotic patients, ulinastatin delays the onset of neuromuscular block produced by vecuronium. After pretreatment with ulinastatin, the speed of recovery from neuromuscular block in patients with cirrhosis becomes similar to that seen in healthy patients. PMID- 11879210 TI - The influence of induction technique on intubating conditions 1 min after rocuronium administration: a comparison of a propofol-ephedrine combination and propofol. AB - We conducted a double blind, prospective, controlled trial comparing intubating conditions after induction with a propofol-ephedrine combination or propofol alone, followed by rocuronium. One hundred adult patients were randomly assigned to receive either propofol 2.5 mg x kg(-1) and ephedrine 15 mg in combination or propofol 2.5 mg x kg(-1) given over 30 s, followed by rocuronium 0.6 mg x kg(-1) given over 5 s. Tracheal intubation was performed 1 min later. Jaw relaxation, vocal cord position and diaphragmatic response were used to assess intubation conditions. Tracheal intubation was successful and acceptable in all patients. There was a significantly higher proportion of intubating conditions graded as 'excellent' in the propofol-ephedrine group (84%) than in the propofol group (32%) (p < 0.0001). Vocal cord position and response to intubation were significantly better in the propofol-ephedrine group, although jaw relaxation was similar. Mean arterial pressure was maintained at pre-induction levels in the propofol-ephedrine group. In conclusion, induction with propofol and ephedrine in combination provided significantly better intubating conditions than propofol alone, when followed by rocuronium. PMID- 11879211 TI - The effects of beta2 adrenoceptor gene polymorphisms on pressor response during laryngoscopy and tracheal intubation. AB - We investigated whether human beta2 adrenoceptor (beta2AR) gene polymorphisms are associated with the pressor response to laryngoscopy and tracheal intubation. Ninety-two patients undergoing elective surgery under general anaesthesia were enrolled into this study. Arterial systolic pressure, heart rate and rate pressure product were measured before induction of anaesthesia and 1 min following laryngoscopy and tracheal intubation. Genomic DNA was then used to identify the beta2AR-16 and beta2AR-27 genes using an allele-specific polymerase chain reaction method. Using multiple linear regression models, controlling for age, sex, weight, baseline blood pressure, heart rate and rate pressure product, we found that patients who possessed the glutamic acid homozygote of beta2AR-27 produced significantly greater changes in mean arterial pressure and rate pressure products than patients with the glutamine homozygote of beta2AR-27 (beta coefficient for mean blood pressure = 11.81, beta coefficient for pulse-pressure product = 8.76, both p-values = 0.023). These findings suggest that genetic variability in the human beta2AR gene polymorphisms may be associated with the pressor response to laryngoscopy and tracheal intubation. PMID- 11879212 TI - The effects of subanaesthetic concentrations of xenon in volunteers. AB - This study reports the subjective, psychomotor and physiological properties of subanaesthetic concentrations of xenon. Ten healthy male volunteers received either xenon or nitrous oxide in a randomised crossover study design. The subjects breathed either xenon (Xe) or nitrous oxide (N2O) from a closed circuit breathing system, according to a randomised, double-blind protocol. The concentration of xenon required to produce sedation, ranged between 27 and 45% (median 35%). All subjects completed the xenon protocol. Subjects were tested using the Critical Flicker Fusion test and derived electroencephalogram parameters, however, neither test was found to reliably predict sedation. The respiratory rate decreased markedly during sedation with xenon. The subjects did not experience any airway irritability (coughing, breath-holding or laryngospasm) during administration of either gas. One subject required anti-emetic treatment in the N2O group compared to none in the Xe group. Eight subjects reported that they found sedation with xenon pleasant and preferable to nitrous oxide. Xenon sedation was well tolerated and was not associated with any adverse physiological effects, however, it was reported to be subjectively dissimilar to nitrous oxide. PMID- 11879213 TI - Propofol effective concentration 50 and its relationship to bispectral index. AB - Sixty unpremedicated healthy adult patients were studied during induction of anaesthesia with intravenous propofol delivered by a 'Diprifusor' target controlled infusion. Bispectral index (BIS) and spectral edge frequency (SEF95) were measured concurrently with the predicted blood and effect site propofol concentrations. Logistic regression was used to calculate the predicted propofol blood and effect site concentrations required to produce unconsciousness and no response to a noxious stimulus in 50% and 95% of patients and to correlate BIS with these end-points. The Diprifusor TCI software produces anaesthesia at consistent target concentrations. Bispectral index correlates well with clinical end-points and may be useful during propofol anaesthesia. PMID- 11879214 TI - Tracheal trauma from percutaneous tracheostomy using the Griggs method. AB - In a safety evaluation study, relative force and distance measurements during percutaneous tracheostomy were recorded using specially monitored Griggs guidewire dilating tracheostomy forceps on 12 cadavers scheduled to undergo postmortem examination the same day. All measurements were recorded in millivolts and were converted to force and distance via appropriate calibration tables. Markedly more force was required for tracheal destruction than for therapeutic tracheal dilation (87.7 N +/- 19 N vs. 31.6 N +/- 17.1 N, p <0.001). Also relatively less force was required for therapeutic tracheal dilation than for dilation of the pretracheal tissues (44.4 N +/- 17.1 N vs. 31.6 N +/- 17.1 N, p <0.05). These results suggest that the Griggs tracheostomy forceps have a reasonable margin of safety in that tracheal destruction is unlikely to occur inadvertently during therapeutic dilation. Second, the dilation of pretracheal tissues can act as useful guide in knowing how much force to apply in a therapeutic dilation. PMID- 11879215 TI - Percutaneous dilational tracheostomy: a safer technique of airway management using a microlaryngeal tube. AB - Management of the airway in an intubated patient during formation of a tracheostomy can be hazardous. The usual method involves withdrawal of the tracheal tube, which has been providing a secure airway, prior to inserting the tracheostomy tube. A method of airway management, using a microlaryngeal tube, has been devised with the aim of maintaining full tracheal intubation and ventilation until the correct position of the tracheostomy tube can be verified. An audit of 250 successive cases of percutaneous tracheostomy demonstrated this method to be safe and effective. PMID- 11879217 TI - Bilateral hypoglossal nerve injury following the use of the laryngeal mask airway. AB - A healthy 54-year-old man undergoing elective knee arthroscopy developed bilateral hypoglossal nerve palsy, lasting 6 weeks following the use of a laryngeal mask airway. He suffered impairment of speech and difficulty in swallowing, the latter resulting in almost 7 kg loss of weight within 2 weeks of surgery. We discuss the possible aetiology of the injury and review the literature describing injuries to the hypoglossal nerve. PMID- 11879216 TI - Evaluation in an anaesthetic simulator of a prototype of a new drug administration system designed to reduce error. AB - Ten anaesthetists were observed while providing anaesthesia for two simulated surgical procedures, twice using conventional methods and twice using a prototype of a new drug administration system designed to reduce error. Aspects of each method were rated by users on 10-cm visual analogue scales (10 being best) and comments were invited. Median safety scores were 7.7 cm (range 4.3-8.9) for the new system and 4.6 cm (1.3-8.2) for conventional methods (p = 0.009). The new system was compared favourably with conventional methods in respect of safety (p = 0.005), clinical acceptability (p = 0.008), organisation and layout (p = 0.047), and acceptability for use on patients (p = 0.005). The new system saved time in the preparation of drugs both before anaesthesia (105 vs. 346 s; p < 0.001) and during anaesthesia (20 vs. 104 s; p < 0.001). Comments facilitated development of the system and the evaluation endorsed proceeding to a clinical trial. PMID- 11879218 TI - Use of methoxamine in the resuscitation of epinephrine resistant electromechanical dissociation. PMID- 11879221 TI - The Airway Management Device (AMD) is not 'reliable and safe'. PMID- 11879222 TI - Nasal endoscopy prior to nasotracheal intubation. PMID- 11879223 TI - A more 'failsafe' approach to difficult intubation with the gum elastic bougie. PMID- 11879224 TI - Capnography for feeding tube placement. PMID- 11879225 TI - A new cause of airway obstruction. PMID- 11879226 TI - A new clinical sign during one-lung anaesthesia: fact or fiction? PMID- 11879227 TI - Airway management of a child with temporomandibular joint ankylosis following otitis media. PMID- 11879228 TI - Outreach critical care services. PMID- 11879229 TI - Adsorption Technologies and Blood Purification Procedures. Proceedings of an international conference in honor of Horst Klinkmann on his 65th birthday. May 5 7, 2000. PMID- 11879230 TI - In celebration of an honored colleague and cherished friend. PMID- 11879231 TI - A laudatio for Horst Klinkmann. PMID- 11879234 TI - Patient safety technology for microadsorbent systems in extracorporeal blood purification. AB - Alternative technologies for extracorporeal blood purification systems based on microadsorbents in suspension are discussed. Principally, microadsorbents offer higher efficiency and flexibility when compared to conventional column-based adsorption systems. Systems already clinically employed (e.g., BioLogic DT) or close to clinical application (e.g., the microspheres-based detoxification system, MDS) are described. The MDS technology, in particular, is characterized by efficiency and a high degree of flexibility with respect to both the use of different adsorbents as well as the combination with hemodialysis/hemofiltration therapy. It was designed for continuous use in intensive-care units, but enables also the removal of low-density lipoprotein, fibrinogen, autoimmune antibodies, immune complexes, and other pathophysiologically relevant substances. Alternative anticoagulation regimes and safety systems on fluorescence sensor technology have recently been developed for the MDS and are presented in this paper. PMID- 11879235 TI - A comparison between epoetin omega and epoetin alfa in the correction of anemia in hemodialysis patients: a prospective, controlled crossover study. AB - The objective of the study was to evaluate and compare the safety and effectiveness of epoetin omega (produced in baby hamster kidney cells) and epoetin alfa (produced in Chinese hamster ovary cells) in sustaining the correction of anemia in maintenance hemodialysis patients. The study, a prospective and controlled crossover, was completed in 38 stable patients treated with both epoetins for 24 weeks. Group A (17 patients) started with epoetin omega, and Group B (21 patients) started with epoetin alfa. After 24 weeks, a 4 week crossover (wash out) was made: Group A was switched to epoetin alfa and group B to epoetin omega for the next test period of 24 weeks. Both epoetins were administered subcutaneously after each dialysis. Doses were adjusted with the aim of maintaining a target hemoglobin level between 10 and 12 g/dl (hematocrit 30% to 35%). The mean weekly dose of epoetin omega/kg body weight (BW) was 67 +/- 43 U. The mean weekly dose of epoetin alfa/kg BW was 86 +/- 53 U. The average of all mean values of hemoglobin (Hb) during treatment with epoetin omega was 11.4 +/- 0.7 g/dl (hematocrit 34 +/- 2%), and during treatment with epoetin alfa was 11.3 +/- 0.7 g/dl (hematocrit 33 +/- 2%) (not significant). Thromboses of vascular access occurred in 3 patients during an epoetin omega treatment and in 3 patients during epoetin alfa treatment. At the site of injection, only 1 patient described a mild pain when treated with epoetin omega and only 6 patients when treated with epoetin alfa. In conclusion, both epoetin omega and epoetin alfa were effective in correcting the anemia of all studied patients. However, lower doses of epoetin omega were needed to maintain the same target hemoglobin level. No serious side effects with either epoetin were noted. The authors believe that additional comparisons of different epoetin preparations should be performed and will provide better insight into their biological activity and clinical responsiveness. PMID- 11879236 TI - Recurrence of left ventricular hypertrophy following cessation of erythropoietin therapy. AB - The high cardiac output state is considered a major factor for occurrence of left ventricular hypertrophy (LVH). Increased left ventricular mass is a powerful predictor of morbidity and mortality. We analyzed morphologic changes of the heart in dialysis patients during treatment with erythropoietin (EPO) and after cessation of therapy. Fourteen hemodialysis patients were treated with EPO for 1 year. They were above age 18, dialyzed 3 times per week, and with a hematocrit below 28 vol%. EPO was given subcutaneously, at a dose of 20 U/kg body weight 3 times per week, before each hemodialysis session. Anemia was corrected and hematocrit maintained between 30 and 35 vol%. When this part of the study was completed, EPO was stopped in all 14 patients. Echocardiography was performed three times: at baseline, at 12 months of therapy, and 1 year after EPO cessation. Mean hematocrit of the group at these 3 time intervals was 23.78 +/- 2.11 vol%; 33.14 +/- 1.95 vol%; and 25.93 +/- 5.23 vol%, respectively (mean +/- SD). The following echocardiographic changes occurred. End-diastolic volume decreased from 134.8 +/- 25.4 to 113.2 +/- 26.4 ml and increased back to 136.2 +/ 46.2 ml. Left ventricular mass decreased from 296.6 +/- 62.4 to 225.2 +/- 52.7 g and increased again to 311.7 +/- 106 g. Cardiac output decreased from 7,295.8 +/- 2,166.9 to 5,816.4 +/- 1,216.2 ml/min and increased to 6,803.2 +/- 1,646.5 ml/min. Total peripheral resistance increased from 1,360.8 +/- 428 to 1,691.3 +/- 326 and decreased again to 1,242.8 +/- 303.3 dyne x s/cm5. All these changes were significant. Mean arterial pressure increased from 114.7 +/- 13.9 to 119.3 +/- 13.8 mm Hg and decreased to 100.5 +/- 9.3 mm Hg. LVH could be affected severely by the degree of anemia in uremics and was reversible. PMID- 11879237 TI - The molecular adsorbents recycling system as a liver support system based on albumin dialysis: a summary of preclinical investigations, prospective, randomized, controlled clinical trial, and clinical experience from 19 centers. AB - Artificial liver support aims to prolong survival time of patients with liver failure by detoxification. Albumin as a molecular adsorbent in dialysis solution is capable of attracting even tightly albumin-bound toxins from blood into the dialysate if a specific dialysis membrane is used and if the albumin's binding sites are on-line-purified by a sorbent/dialysis-based recycling system (i.e., molecular adsorbents recycling system, or MARS). The MARS technology has been shown to remove water-soluble and albumin-bound toxins and to provide renal support in case of renal failure. Fourteen centers have reported that MARS treatment improved mental status of patients with liver failure and hepatic encephalopathy. In treating liver failure and cholestasis, MARS was associated with hemodynamic stabilization, improvement of hepatic and kidney function, and disappearance of pruritus. In hepatic failure and hepatorenal syndrome, a prospective, randomized, controlled trial of MARS treatment was able to prolong survival time significantly. MARS has been used in 26 patients with acute liver failure or primary graft dysfunction. Nineteen centers reporting on 103 patients have shown that MARS treatment is safe, easy to handle, feasible, and effective. PMID- 11879238 TI - MTS colorimetric assay in combination with a live-dead assay for testing encapsulated L929 fibroblasts in alginate poly-L-lysine microcapsules in vitro. AB - Biomaterials such as applied in microcapsules may have harmful effects on encapsulated cells. Up to now, there are no adequate assays available for testing the function and viability of cells in capsules. Therefore, we investigated whether the combination of MTS proliferation assay and live-dead viability assay is suitable for testing microencapsulated L929 fibroblasts in long-time culture. Proliferation of L929 cells was shown by a significant increase of formazan absorbance within the first 3 weeks (Day 0: 0.132 +/- 0.047; Day 7: 0.404 +/- 0.101; Day 14: 0.728 +/- 0.239; Day 21: 0.877 +/- 0.224) followed by stagnation and decrease thereafter. This was confirmed by an increasing proportion of dead cells measured by the live-dead assay. Thus, proliferation of encapsulated L929 can be reliably investigated by the MTS assay. In combination with life-dead assays, the proliferation can be correlated to the survival rate of the encapsulated cells. PMID- 11879239 TI - Selective hemapheresis, an effective new approach in the therapeutic management of disorders associated with rheological impairment: mode of action and possible clinical indications. AB - The in vitro measurement of whole-blood viscosity, plasma viscosity, and erythrocyte aggregability is easy to perform, but they only allow a partial insight into the complexity of blood flow characteristics; however, they permit definition of the rheological properties of new hemorheological therapeutic modalities such as extracorporeal plasma therapy as described in this paper. Under more theoretical aspects, it becomes obvious that such hemorheological approaches should either improve the vasomotoric properties of blood vessels, reduce the circulating red blood cell concentration, or improve the viscosity by reducing the concentration of hemorheologically relevant plasma proteins. In this review, the rheological effect of a single apheresis treatment with different devices was compared. Due to their differences in selectivity, the extracorporeal methods have different effects on the rheologically relevant plasma proteins, and, therefore, their rheological effectiveness differs remarkably. Today, the classical blood letting and plasma exchange treatment have been replaced by erythrapheresis and selective devices for extracorporeal plasma treatment, respectively. For more than 10 years, the following 5 more-or-less selective apheresis procedures are commercially available: immunoadsorption, differential filtration, polyanion adsorption by dextrane sulfate as well as by polyacrylate, and polyanion precipitation by heparin as polyanion. The last three procedures are semiselective and, therefore, relatively unspecific whereas immunoadsorption only affects the plasma lipoprotein concentration. Several studies have shown the effective use of extracorporeal hemorheotherapy for the treatment of various diseases including macro- and cryoglobulinemia, Raynaud's disease, hyperlipoproteinemia (often characterized by premature atherosclerosis and coronary heart disease and peripheral arterial occlusive disease), cerebral multi infarct demention and acute ischemic stroke, sudden hearing loss, and acute occlusion of the central retinal artery. PMID- 11879240 TI - Selective removal of circulating immune complexes from patient plasma. AB - The principle of a patient-specific immunoadsorber (PsIA) is demonstrated. Studies with model systems (HSA/anti-HSA) on immobilization, stability, and leakage form the basis for the presented fast-performance liquid chromatography (FPLC) and batch experiments, which were conducted using two different protein A adsorbers and autologous and heterologous PsIA systems. Experiments to determine the binding capacity of protein A adsorbers and PsIAs are described. In all experiments, the adsorption of plasma IgG, total protein, and C1q and C3d circulating immune complexes were measured. Plasma of patients with autoimmune diseases (rheumatoid arthritis, systemic lupus erythematosus) was investigated. Analysis was performed in both the initial plasma and the flow-through or supernatant. Results of the investigations using FPLC and batch experiments were compared. Autologous PsIA systems are suitable for the selective removal of elevated levels of circulating immune complexes in the plasma. PMID- 11879241 TI - Low-density lipoprotein apheresis: clinical results with different methods. AB - In 40 patients (22 women, 18 men) suffering from familial hypercholesterolemia resistant to diet and lipid lowering drugs, low-density lipoprotein (LDL) apheresis was performed over 84.9 +/- 43.2 months. Four different systems (Liposorber, 28 of 40, Kaneka, Osaka, Japan; Therasorb, 6 of 40, Baxter, Munich, Germany; Lipopak, 2 of 40, Pocard, Moscow, Russia; and Dali, 4 of 40, Fresenius, St. Wendel, Germany) were used. With all methods, average reductions of 50.6% for total cholesterol, 52.2% for LDL, 64.3% for lipoprotein (a) (Lp[a]), and 43.1% for triglycerides, and an average increase of 10.3% for high-density lipoprotein (HDL) were reached. Severe side effects such as shock or allergic reactions were very rare (0.5%) in all methods. In the course of treatment, an improvement in general well being and increased performance were experienced by 39 of 40 patients. Assessing the different apheresis systems used, at the end of the trial, there were no significant differences with respect to the clinical outcome experienced with the patients' total cholesterol, LDL, HDL, and triglyceride concentrations. However, to reduce high Lp(a) levels, the immunoadsorption method with special Lp(a) columns (Lipopak) seems to be most effective: -59% versus -25% (Kaneka) - (Baxter), and -29% (Dali). The present data demonstrate that treatment with LDL apheresis of patients suffering from familial hypercholesterolemia resistant to maximum conservative therapy is very effective and safe even in long term application. PMID- 11879242 TI - Changes in lymphocytic cluster distribution during extracorporeal immunoadsorption. AB - The success of apheresis treatment is often measured as a decrease in the detected antibodies and an improvement in different disease-related scores. Sometimes, however, the seriousness of the disease does not correlate with the antibody level. During a period of 8 years, 15 patients (3 myasthenia gravis, 1 multiple sclerosis, 2 systemic lupus erythematosus, 3 alloimmunized kidney transplant, 6 rheumatoid arthritis) were treated by protein A immunoadsorption. Lymphocyte subpopulations (activated T cells, cytolytic T cells, B cells, natural killer cells) and inflammatory proteins (ferritin, C-reactive protein, alpha1 antitrypsin, alpha2-macroglobulin) were analyzed. After observing clinical outcomes, the patients could be divided into 2 groups, respectively: Group 1, responding patients with remission of disease; and Group 2, delayed-responding patients, who required chronic treatment. Group 1 patients characteristically showed a greater increase in activated T and cytolytic T cells which correlated with a greater decrease of B cells. It might be possible that protein A immunoadsorption induced immunomodulation. Further immunological investigation is required to verify these findings. PMID- 11879243 TI - RheoSorb: a specific adsorber for fibrinogen elimination in clinical situations with impaired rheology. AB - A functional microcirculation is crucial for the normal function of an organism. In many physiopathological situations, impaired microcirculation may contribute to the development or progress of diseases. Microcirculation is closely interrelated with blood and especially plasma rheology. Thus, improvement of plasma viscosity has beneficial effects on rheology, microcirculation, and the related tissue microenvironment. However, at best tools that only have a minor influence on plasma viscosity exist so far. Fibrinogen is known to be the major contributor to plasma viscosity, making it an interesting target for therapeutic intervention. An adsorber specific for fibrinogen was developed on the basis of the TheraSorb technology. The TheraSorb technology (PlasmaSelect AG, Teterow, Germany) allows the selective removal of components from human blood plasma by means of an affinity chromatography column. A ligand specific for a defined plasma component is coupled to a solid matrix (sepharose) thus binding and eliminating the target molecule from plasma. Using a fibrinogen specific pentapeptide as ligand, selective removal of fibrinogen, fibrin, and degradation products, containing the target sequence of these molecules, can be obtained. The LIFE-18, a state-of-the-art integrated plasma therapy instrument, is used to perform the treatment. The procedure improves plasma and whole blood viscosity in a dose dependent manner as shown in Phase 1 and 2 clinical trials. This article describes the first clinical experience in patients with diabetic foot syndrome and provides an outlook for further clinical and scientific investigations related to this promising new procedure. PMID- 11879244 TI - Quo vadis dialysis membrane? AB - The development of dialysis membranes is closely related to the development of dialysis as a routine therapy for patients with kidney failure. Without having membranes and dialyzers available as commodity products, the treatment of more than 1 million uremic patients worldwide would be impossible. Several transition periods can be identified: a change in membrane geometry from flat sheet to capillaries, a shift in market appreciation from cellulose to synthetic polymers, and from low-flux to high-flux dialyzers. This shift is supported by the notion that convective therapies using high-flux membranes allow the removal of large molecular-weight solutes. From a historical background, three eras of perception can be identified for both membrane and dialysis development. First, the period of survival when nephrologists had to focus on techniques for blood access and availability of membranes. Second, the period of issues dedicated to rather specific features of membranes and dialysis therapy such as dose of dialysis, reuse, sterilization, and membrane biocompatibility. And third, the period of quality tops this sequence with a complicated approach: the principal area of interest from the medical community has switched to issues such as quality of life, morbidity, mortality, therapy standards, and cost-effectiveness. New membrane developments should focus on this situation. PMID- 11879245 TI - Alternative timeframes for hemodialysis. AB - Routinely, maintenance hemodialysis is applied according to an alternate day time schedule with dialysis sessions lasting 3 to 5 h. In analogy with continuous ambulatory peritoneal dialysis, a more gradual and more consistent removal pattern could be achieved by modifying this timeframe, e.g., by applying daily hemodialysis and/or by prolonging hemodialysis sessions to 8 h or more. Recently, a number of dialysis units implemented such alternative timeframes. Both the clinical and metabolic status showed dramatic improvements with beneficial effects on blood pressure, anemia, and potassium and phosphate levels. These changes occurred despite decreases in the dosage of the drugs administered to cope with these problems. However, controlled studies comparing morbidity and mortality of these alternative timeframes to the more traditional approaches are lacking at this moment. PMID- 11879246 TI - Continuous renal replacement therapy and plasma exchange in newborns and infants. AB - The objective of our study was to present our experience in the treatment of small children with continuous renal replacement therapy (CRRT) and plasma exchange (PE). From March 1986 to April 2000, 21 critically ill children (14 newborns and 7 infants) with acute renal failure (ARF) and multiple organ failure were treated with CRRT and PE. In the newborn group, there were 8 males and 6 females, age 15.7 +/- 11.7 days, with body weights of 3,348 +/- 585 g. In the infant group, there were 4 males and 3 females, age 118 +/- 67 days, with body weights 5,186 +/- 734 g. The indications for the beginning of CRRT and/or PE were ARF with anuria and hyperhydration (17 patients), azotemia and anuria (1 patient), hemolytic uremic syndrome (1 patient), and hyperammonemia (2 patients). In all patients, peritoneal dialysis was considered inappropriate. PE and CRRT monitors were used, double lumen 5 Fr and 7 Fr hemodialysis catheters were the vascular access, low dose heparin and prostacyclin were anticoagulants, and lactate or bicarbonate buffered replacement solutions were used predilutionally. Side events were clotting within the extracorporeal circuit, catheter malfunction, serious hypotension (6 patients), and pulmonary edema (1 patient). Ten of 21 patients (47.6%) recovered renal function and 9 of 21 patients (42.9%) survived. Survivors had fewer failing organs (3.6 +/- 0.5) than nonsurvivors (4.8 +/- 0.9) (p = 0.0008). Pump driven CRRT and PE were feasible, efficient, and safe procedures in newborns and infants. Without CRRT, it is uncertain whether any of our patients would have the chance to survive. PMID- 11879247 TI - Online hemodiafiltration versus acetate-free biofiltration: a prospective crossover study. AB - Online hemodiafiltration (online HDF) and acetate-free biofiltration (AFB) are 2 innovative renal replacement therapies. Convincing evidence has shown that both techniques are superior to conventional hemodialysis in many aspects. The aim of the present investigation was to compare online HDF and AFB in 12 stable maintenance hemodialysis patients in a prospective, randomized crossover trial. Twelve stable dialysis patients, age 49.7 +/- 11.3 years and on dialysis for 83.5 +/- 76.7 months, were treated prospectively and randomly by either AFB, predilution HDF (pre-HDF), or postdilution HDF (post-HDF) for a total of 36 weeks using exclusively F60S high-flux dialyzers. Routine blood biochemical tests, bone metabolism parameters, and clearance for both small and larger molecular weight substances were measured at defined intervals. During the trial period inter- and intradialysis symptoms, e.g., hypotensive episodes and intradialysis arterial blood gas analyses, were recorded. Both online HDF and AFB were well accepted by the overwhelming majority of patients and also by the dialysis staff. Pretreatment sodium, total and ionized calcium, chloride, bicarbonate, and urea did not differ within or between the 3 treatment groups. Potassium increased slightly in HDF patients while phosphate and beta2-microglobulin (beta2-M) decreased in all groups. After dialysis, AFB patients exhibited a significantly higher bicarbonate concentration and lower potassium level when identical potassium concentrations in dialysate were used. Patients receiving AFB manifested less intradialysis partial pressure of oxygen drop and partial pressure of carbon dioxide rise than those on HDF treatments. HDF treatments could afford higher single-pool and double-pool Kt/V, higher effective urea and beta2M clearance, and lower total interdialysis symptom scores than the AFB treatment method. While bone metabolism parameters did not differ between the 3 dialysis modalities, some parameters such as deoxypyridinoline in HDF and osteocalcin, pyridinoline, and deoxypyridinoline in AFB deteriorated at the end of the crossover study. Aluminum concentration decreased progressively to about one-third of prestudy values at the end of the study with all 3 treatments. AFB was associated with a lower predialysis mean arterial pressure (MAP), a smaller drop in MAP during treatment, and similar hypotension episodes compared with the 2 HDF treatments. Albumin concentration showed a trend to decrease during the first 2 months of the trial period followed by a slight increase thereafter but still significantly lower than initial value at the end of crossover. Both online HDF and AFB share most of the features of optimal renal replacement therapy. Online HDF is superior to AFB in such aspects as increased delivered dialysis dose both for small and larger molecular weight toxins and less interdialysis symptoms. On the other hand, AFB is associated with a smaller effect on arterial blood gas values and improved intradialysis hemodynamic tolerance. Some dialysis related symptoms and complications in the case of our AFB practice could be attributable, at least in part, to low dialysate calcium level. PMID- 11879248 TI - Does treatment modality have an impact on anemia in patients with chronic renal failure? Effect of low- and high-flux biocompatible dialysis. AB - Renal anemia significantly affects the morbidity and mortality of dialysis patients. The aim of the present study was to establish whether the severity of anemia and success of its treatment differs when using high-flux (HF) or low-flux (LF) hemodialysis. Twenty-five patients on long-term hemodialysis with a mean hematocrit (Hct) of 33%, S alb of 36 g/L, and Kt/V urea of 1.5 were divided into Groups X (n = 13) and Y (n = 12) in a prospective randomized crossover study. Group X was initially treated with LF hemodialysis to be followed by HF hemodialysis for 8 weeks each time. Group Y started with 8 week HF hemodialysis and continued, after crossover, with 8 weeks of LF hemodialysis. HF and LF hemodialysis were performed with polysulfone dialyzers F 7HPS and F60S (Fresenius Medical Care, Bad Homburg, Germany). Hct values, examined at 2 week intervals, did not differ significantly between Groups X and Y during 16 weeks of the study at any measuring interval. In another analysis, all results of HF hemodialysis (first 8 weeks of Y + second 8 weeks of X) were pooled as were all data of LF hemodialysis (first 8 weeks of X + second 8 weeks of Y). No significant relationships were noted between duration of treatment with HF hemodialysis and Hct (n = 72, rS = 0.11, p = 0.36) and between duration of LF hemodialysis and Hct (n = 74, rS = 0.02, p = 0.87) in the newly formed groups. The Hct measured during HF hemodialysis and LF hemodialysis did not differ significantly. Analysis of variance did not reveal a relationship between Hct and the HF or LF membrane. As HF and LF hemodialysis differed significantly in Kt/V urea, the variables were adjusted to identical Kt/V urea using analysis of covariance. No relationship between Hct and dialysis membrane permeability was demonstrated even in this case. Use of biocompatible LF and HF membranes in standard hemodialysis modes under conditions of adequate dose of dialysis and the time period studied did not result in different effects on anemia. PMID- 11879249 TI - Polyetherimide: a new membrane-forming polymer for biomedical applications. AB - Membranes for biohybrid organs such as the biohybrid liver support system have to face 2 different environments, namely blood and tissue cells. Accordingly, the respective membrane surfaces must have optimal properties in terms of biocompatibility for blood or tissue. Flat membranes prepared by a phase inversion process from polyetherimide were modified by binding of tris (hydroxymethyl)-aminomethane to obtain a surface with hydroxyl groups by binding of polyethylene imine to attach a hydrophilic macromolecule with amine groups useful as a spacer for later bonding of further ligands and by attachment of heparin. The binding of the different ligands was successful as monitored by different physicochemical methods. The blood response of plain polyetherimide was comparable to that of polyacrylonitrile, and it could be further improved by the binding of heparin. The tissue compatibility of polyetherimide and its different modifications was compared with commercial cell culture substrate membranes (Millicell) and found to be comparable for polyetherimide and even better after the modification with tris-(hydroxymethyl)-aminomethane. In conclusion, polyetherimide seems to be an interesting material for the production of membranes for application in biohybrid organ systems. PMID- 11879250 TI - Development of cellulose-DNA immunoadsorbent. AB - The aim of this study was to prepare a DNA immunoadsorbent for the specific, extracorporeal removal of anti-DNA antibodies from the blood of patients with systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA). Two kinds of cellulose beads were applied as a carrier. Calf thymus DNA was covalently coupled to the carrier using the epichlorohydrin method. Efforts were focused on optimization of conditions for activation and coupling, trying to couple as much DNA as possible to a certain amount of carrier. It was found that the activation level increased with the increase of NaOH concentration and the amount of epichlorohydrin used. The mole of epichlorohydrin must be in excess of that of NaOH because excess NaOH could react further with the epoxy groups in the beads resulting in a decrease of activation level. High activation level could be obtained in a medium of 3.0 M NaOH. The DNA coupling was found to be mainly temperature and pH dependent. Using 0.1 M Tris-HCl buffer, pH 8 at a temperature of 50-90 degrees C, more than 3 mg of DNA could be coupled to 1 ml of wet beads. Prolonging the coupling reaction under 50 degrees C to 72 h resulted in the same coupling capacity as that obtained under 90 degrees C. To evaluate the adsorption ability for anti-DNA of this immunoadsorbent, batch and circulation tests were applied using SLE patient plasma. The immunoadsorbents showed excellent adsorption capacity, especially the cellulose with smaller size (200-300 microm). The incubation of 20 ml of patient's plasma with 1 ml of adsorbent resulted in an 80% decline in the anti-DNA antibody level. In the circulation tests, 30 ml of plasma was circulated through a column containing 3 ml of adsorbent. The maximum decline in anti-DNA level, 80%, was obtained after 60 min. Such high adsorption capacity and high adsorption rate suggest this immunoadsorbent may be used for treatment. For comparison, 1,4-butanediol diglycidyl ether activation method and other DNA sources were tested with the same protocol. PMID- 11879251 TI - Impregnation of the the polymeric graft with adhesives molecules, typically oligopeptides or glycoprotein improves retention. PMID- 11879254 TI - Pharmacokinetics of sildenafil after single oral doses in healthy male subjects: absolute bioavailability, food effects and dose proportionality. AB - AIMS: To determine the absolute bioavailability, dose proportionality and the effects of food on the pharmacokinetics of single oral doses of sildenafil citrate. METHODS: Three open-label, randomized crossover studies were conducted in healthy male subjects. Absolute bioavailability was determined by comparing pharmacokinetic data after administration of single oral and intravenous 50-mg doses of sildenafil (n=12 subjects). Food effects were examined by comparing pharmacokinetic data for sildenafil and its primary circulating metabolite, UK 103,320, after administration of a single oral 100-mg dose in the fasted and fed states (n=34 subjects). Dose proportionality was assessed from pharmacokinetic data obtained after administration of four single oral doses of sildenafil (25, 50, 100 and 200 mg) to 32 subjects. The safety and tolerability of sildenafil were also assessed in all of these studies. RESULTS: The calculated absolute oral bioavailability of sildenafil was 41% (90% CI: 36--47). Food slowed the rate of absorption, delaying mean tmax by approximately 1 h and reducing Cmax by 29% (90% CI: 19--38). Systemic exposure, as assessed by the mean area under the plasma concentration--time curve (AUC), was reduced by 11% (90% CI: 6--16). These food effects were not considered to be of clinical significance. There was statistical evidence of nonproportionality in Cmax and AUC over the dose range 25--200 mg. However the degree of nonproportionality was small, with predicted increases in Cmax and AUC of 2.2- and 2.1-fold, respectively, for a doubling in dose, and was thought to be clinically nonsignificant. Sildenafil was well tolerated in the three studies; the majority of adverse events were mild and transient. CONCLUSIONS: Sildenafil had a mean absolute bioavailability of 41%. Food caused small reductions in the rate and extent of systemic exposure; these reductions are unlikely to be of clinical significance. Across the dose range of 25--200 mg, systemic exposure increased in a slightly greater than dose-proportional manner. PMID- 11879255 TI - Comparative human pharmacokinetics and metabolism of single-dose oral and intravenous sildenafil. AB - AIMS: To characterize the absorption, metabolism and excretion of an oral and intravenous (IV) dose of radiolabelled [14C]-sildenafil citrate in healthy male subjects. Specific objectives were to measure the cumulative amount of drug related radiolabelled material excreted in the urine and faeces to characterize urinary and faecal radioactivity as unchanged sildenafil or its metabolites, and to quantify blood and plasma total radioactivity and unchanged drug concentrations. METHODS: Six healthy male subjects between the ages of 45 and 58 years were enrolled in an open-label, parallel-group study; three subjects received the oral dose and three received the IV dose. Oral drug was administered as a single dose of 50-mg [14C]-sildenafil, and IV drug was administered as a single dose of 25-mg [14C]-sildenafil infused over 25 min. Each dosage form contained 50 microCi of radioactivity. For radioactivity assays, whole blood, plasma, urine and faeces samples were taken predose and at specified intervals up to 5 days postdose. Plasma samples were assayed for sildenafil and the metabolites UK-103,320 and UK-150,564. Metabolite profiling was also performed in plasma, faeces and urine. RESULTS: Absorption of sildenafil after oral administration was rapid and approximately 92% whilst the absolute bioavailability was limited to 38%, due to first-pass metabolism. Mean AUCt values showed that sildenafil accounted for about 60% of the total circulating radioactivity in the plasma after IV administration and for 32% after oral administration. Concentrations of radioactivity in whole blood were lower than in plasma, indicating limited penetration of sildenafil into blood cells. No unchanged sildenafil was detected in either urine or faeces, demonstrating that metabolism was the major mechanism of drug clearance. The principal routes of metabolism were N-demethylation, oxidation and aliphatic dehydroxylation. Sildenafil was well tolerated, with treatment-related adverse events reported by three subjects. Two of these were mild, and there was one case of moderate leg pain. CONCLUSIONS: The pharmacokinetics of radiolabelled [14C]-sildenafil were consistent with rapid absorption, first-pass metabolism and primarily faecal elimination of N-demethylated metabolites. PMID- 11879256 TI - The effects of age and renal and hepatic impairment on the pharmacokinetics of sildenafil. AB - AIMS: To investigate the effects of age and renal and hepatic impairment on the pharmacokinetics, tolerability and safety of sildenafil (single 50-mg oral dose) and its major circulating N-desmethyl metabolite, UK-103,320. METHODS: Three open label, parallel-group studies were conducted. The first study compared sildenafil pharmacokinetics, safety and toleration in 15 healthy young male subjects (mean age 30 years; range 19--45 years) to 15 healthy elderly male subjects (mean age 70 years; range 65--81 years). The second study included eight male volunteers with normal renal function and 16 male volunteers with varying degrees of renal impairment as assessed by measurement of creatinine clearance (CLcr). The third study included 12 male volunteers with normal hepatic function and 12 male volunteers with chronic stable hepatic cirrhosis (Child-Pugh A and B). For all three studies, blood and urine samples were collected predose and at specified intervals up to 48 h postdose for assays of sildenafil and UK-103,320, and measurements of protein binding. RESULTS: Significant differences in Cmax and AUC were observed between the young and the elderly subjects for both the parent drug and the metabolite. In the elderly, AUC values were approximately twice as high and Cmax values 60--70% higher than those for young men, while t1/2 values were approximately 1 h longer for sildenafil and 2 h longer for UK-103,320. Due to a significantly smaller unbound fraction of drug in the elderly, free drug concentrations were only approximately 40% higher in the elderly group compared to the young group. In the renal impairment study, significant correlations with CLcr were demonstrated for sildenafil oral clearance (CL/F) and Cmax and UK 103,320 Cmax and AUC. Pairwise comparisons between subjects with normal renal function and those with severe renal impairment (CLcr<30 ml min-1) supported these findings, showing significant increases in Cmax and AUC for both the parent drug and the metabolite in the severely impaired subjects. The hepatic impairment study demonstrated that the pharmacokinetics of sildenafil were altered in subjects with chronic stable cirrhosis, as shown by a 46% reduction in CL/F and a 47% increase in Cmax compared with subjects with normal hepatic function, suggesting a reduction in first-pass metabolism as well as systemic clearance. The increase in systemic exposure for UK-103,320 was approximately twice that seen for the parent drug. In all three studies, sildenafil was well tolerated, most adverse events were mild and no subjects discontinued treatment. CONCLUSIONS: Sildenafil pharmacokinetics were affected by age and by renal and hepatic impairment, suggesting that a lower starting dose of 25 mg should be considered for patients with severely compromised renal or hepatic function. PMID- 11879258 TI - The effects of steady-state erythromycin and azithromycin on the pharmacokinetics of sildenafil in healthy volunteers. AB - AIMS: Sildenafil, an effective oral treatment for erectile dysfunction, is predominantly metabolized by the cytochrome P450 isozyme 3A4, which is inhibited by a number of the macrolide antibiotics. Therefore, two placebo-controlled, parallel-group studies were conducted to evaluate the effects of multiple doses of erythromycin and azithromycin on the pharmacokinetics, safety and tolerability of a single oral 100-mg dose of sildenafil. METHODS: In the erythromycin interaction study, 26 male volunteers (18--45 years of age) received open-label sildenafil 100 mg on day 1. Half received blinded erythromycin (500 mg) twice daily on days 2--6, and the other half received placebo. On day 6, all subjects received a second 100-mg dose of sildenafil. In the azithromycin interaction study, 24 male volunteers (19--33 years of age) received open-label 100 mg sildenafil on day 1. Half then received blinded azithromycin (500 mg) once daily on days 2--4, and the other half received placebo. On day 4, all subjects received another 100-mg dose of sildenafil. In both studies, blood samples were collected on the first and last study day for the analysis of plasma concentrations of sildenafil and its primary metabolite, UK-103,320. RESULTS: Repeated dosing with erythromycin caused statistically significant increases in the AUC and Cmax of sildenafil (2.8-fold and 2.6-fold, respectively) but had no effect on Tmax, kel or t1/2. A statistically significant 1.4-fold increase in the AUC of UK-103,320 was also observed, as well as a significant decrease in kel, resulting in an increase of about 1 h in t1/2. In contrast, repeated dosing with azithromycin caused no significant change in any pharmacokinetic parameter of either sildenafil or UK-103,320. Erythromycin, azithromycin and sildenafil were well tolerated; adverse events were mild and transient. No subject withdrew from either trial for any reason related to study drug. CONCLUSIONS: These results indicate that erythromycin modifies the pharmacokinetics of sildenafil by inhibiting its CYP3A4-mediated first-pass metabolism. Given these data, a lower starting dose of sildenafil (25 mg) may be considered for patients receiving erythromycin or other potent CYP3A4 inhibitors. Azithromycin did not affect the pharmacokinetics of sildenafil; therefore, no adjustment in dosage is necessary for patients receiving these drugs concomitantly. PMID- 11879257 TI - The effects of cimetidine and antacid on the pharmacokinetic profile of sildenafil citrate in healthy male volunteers. AB - AIMS: To examine the effect of concomitant cimetidine or antacid administration on the pharmacokinetic profile of sildenafil citrate in healthy male volunteers in two open-label, randomized studies. METHODS: The first study was a parallel group design in which 22 healthy male volunteers received sildenafil (50 mg) on days 1 and 5 and cimetidine (800 mg) or placebo on days 3, 4, 5, and 6. Blood samples were collected predose and at specified times up to 48 h postdose on days 1 and 5 to determine plasma levels of sildenafil and its metabolite, UK-103,320. The second study was a two-way crossover design in which 12 volunteers received sildenafil with or without a 30-ml dose of a magnesium hydroxide/aluminium hydroxide antacid. Blood samples were collected and analysed as in the first study. The two study periods were separated by at least 14 days. RESULTS: Coadministration of cimetidine had no statistically significant effect on the tmax or kel of sildenafil but caused a statistically significant increase in sildenafil AUCt and Cmax of 56% and 54%, respectively (P<0.01). Differences between the two treatment groups were smaller for the metabolite than for sildenafil, although cimetidine treatment did significantly (P<0.05) increase the AUCt for UK-103,320 by 30%. Antacid coadministration had no statistically significant effect on any pharmacokinetic parameter of sildenafil or UK-103,320. Whether taken alone, with cimetidine, or with an antacid, sildenafil was well tolerated. Most adverse events were mild in nature, and no subject withdrew from either study for any reason related to the drug. CONCLUSIONS: Cimetidine co administration produced an increase in sildenafil plasma levels; however, this increase is not sufficient to warrant dosage adjustment of either drug. Antacid coadministration had no effect on the pharmacokinetic profile of sildenafil. PMID- 11879259 TI - A population pharmacokinetic analysis of sildenafil citrate in patients with erectile dysfunction. AB - AIMS: To analyse the pharmacokinetics of sildenafil citrate in patients with erectile dysfunction in order to characterize covariate relationships and assist in the development of rational dosage strategies. METHODS: A population pharmacokinetic sampling strategy was incorporated into five phase III clinical study protocols. Overall, 2077 patients, 1335 of whom received sildenafil, were asked to take an additional dose of study drug before their scheduled clinic visits on four or five occasions throughout the study duration. A single plasma sample was obtained at random times postdose (range 1--7 h), and a total of 4582 samples were assayed (average 3.4 samples per individual). RESULTS: For the population average patient (age 58 years; aspartate transaminase [AST], 24 IU l( 1); weight, 87 kg; not receiving CYP3A4 potential inhibitors), typical values for sildenafil (mean +/- SE) were 58.5 +/- 1.4 l h(-1) for apparent clearance (CL/F), 310 +/- 6.92 l for volume of distribution (V/F), and 2.6 +/- 0.176 h(-1) for first-order absorption constant (ka). The value for ka is associated with meal consumption within 2 h predose, at all other times ka was equivalent to an instantaneous bolus administration. The interindividual variabilities were 29% for CL/F, 20% for V/F, and 210% for ka. Over a dose range of 25--100 mg sildenafil, the pharmacokinetics exhibited dose proportionality. There was evidence of nonproportionality (40% increase on average) in relative bioavailability with respect to the 200-mg dose (P<0.001) relative to the other doses. Age, AST concentration, and co-administration with CYP3A4 potential inhibitors significantly influenced CL/F of sildenafil (P<0.001, for each relationship). For age and AST, the extent of the linear relationships (extrapolated from population average values) included a 4% decrease in CL/F for every decade increase and a 6% decrease in CL/F for every 10-unit increase, respectively. Following co-administration of CYP3A4 potential inhibitors, a 14% decrease in CL/F was estimated. Only body weight was found to significantly (P<0.001) influence V/F (a 6% increase in V/F for every 10-kg increase). CONCLUSIONS: The pharmacokinetics of, and covariate influences on, sildenafil in patients with erectile dysfunction were shown to be consistent with those demonstrated in phase I volunteer studies. PMID- 11879260 TI - The effects of sildenafil on human sperm function in healthy volunteers. AB - AIMS: This double-blind, randomized, four-period, two-way crossover study was conducted to evaluate the acute effects of oral sildenafil (100-mg single dose) on sperm motility, count, density, morphology and vitality as well as ejaculate volume and viscosity in healthy male subjects. The concentrations of sildenafil and its primary circulating metabolite UK-103,320 were measured in ejaculate and compared with those in plasma. The study also included assessments of safety and tolerability. METHODS: A total of 17 healthy male volunteers aged 19--34 years were randomized to receive a single 100-mg dose of sildenafil for two periods and a single dose of placebo for two periods, with each period separated by a minimum of 5--7 days. Sperm and ejaculate properties were evaluated from semen samples taken at screening and 1.5 h after dose. An additional semen sample was collected 4 h after dose, and drug and metabolite concentrations were measured in this sample and the sample taken 1.5 h after dose for comparison with plasma concentrations. Blood samples were collected before each dose and 0.25, 0.5, 1, 2, 3, 4 and 6 h after dose for measurement of sildenafil and metabolite concentrations. RESULTS: Sildenafil had no statistically significant effect on sperm motility, count or density; the percentage of abnormal sperm forms; or the percentage of living sperm. It also did not affect ejaculate volume or viscosity. All measures were within normal ranges. Sildenafil distributed into the semen rapidly, resulting in significant correlations between concentrations of sildenafil in the semen and total (R2=0.588) or free (R2=0.454) plasma concentrations (P<0.0001). Total semen concentrations of sildenafil were 18% of total plasma concentrations. UK-103,320 appeared to distribute more slowly from the plasma into the semen, resulting in a lack of correlation between semen and plasma concentrations. The amount of sildenafil and UK-103,320 in the ejaculate was small (< 2x10(-4)% of the administered dose at 1.5 h). Sildenafil was well tolerated; no patient withdrew from the study due to adverse events attributed to sildenafil. CONCLUSIONS: These results indicate that a single 100-mg oral dose of sildenafil does not have an adverse effect on sperm function or ejaculate quality. PMID- 11879262 TI - Flow cytometry and the study of cerebrospinal fluid in leukaemic patients: additional facts. PMID- 11879261 TI - Onset and duration of action of sildenafil for the treatment of erectile dysfunction. AB - AIMS: To determine the onset and duration of action of sildenafil in patients with erectile dysfunction (ED). METHODS: Two randomised, double-blind, placebo controlled, two-way crossover studies were conducted in men with ED of no known organic cause. Study I: The time to onset of erections after sildenafil (50 mg) or placebo dosing following visual sexual stimulation (VSS) was assessed in 17 patients. Patients not achieving >60% penile rigidity by 70 min postdose as measured by a RigiScan(R) monitoring device were assigned an onset time of 70 min. Study II: The duration of grade 3 (hard enough for penetration) and grade 4 (fully hard) erections, determined by self-assessment during 60 min of VSS starting 2 and 4 h after sildenafil (100 mg) or placebo dosing, was measured in 16 patients. RESULTS: Study I: The median time (range) to onset of erections was 27 min (in a range of 12--70) after receiving sildenafil 50 mg. In the sildenafil group, 71% of patients experienced onset of erections within 30 min of dosing, and 82% responded within 45 min. Of the patients who achieved >60% penile rigidity after sildenafil, 86% had done so by 30 min after dosing. Study II: When VSS began 2 h postdose, the median duration of grade 3 or 4 erections was 19.5 min (0--55) for sildenafil vs 0 min (0--23) for placebo. When VSS began 4 h postdose, the median duration was 5 min (0--45) for sildenafil compared with 0 min for placebo (0--27). CONCLUSIONS: Sildenafil is an effective oral treatment for ED that produces a penetrative erection as early as 12 min and for most patients, within 30 min after dosing, and a duration of action lasting at least 4 h. PMID- 11879263 TI - Reversible abnormal chromatin clumping in granulocytes from six transplant patients treated with mycophenolate mofetil: a rare adverse effect mimicking abnormal chromatin clumping syndrome. PMID- 11879266 TI - Stepping up psychosocial care in cancer care. PMID- 11879264 TI - Restoration of erythropoiesis by rituximab in an adult patient with primary acquired pure red cell aplasia refractory to conventional treatment. PMID- 11879267 TI - Issues in bilateral prophylactic mastectomy. PMID- 11879268 TI - Physical functioning and depression among older persons with cancer. AB - PURPOSE: The purpose of this study was to help identify factors to assess which elderly patients are likely to experience problems with physical and psychological functioning in association with cancer or its treatment. DESCRIPTION OF STUDY: A study was undertaken with a sample of 420 patients with cancer who were between the ages of 65 and 98 years and had received an incident diagnosis of breast, colon, lung, or prostate cancer. An analysis of covariance technique was used to determine how cancer site, treatment type, stage of disease, gender, age, comorbidity, symptom severity, and pre-diagnosis levels of physical functioning were related to physical functioning deficit, and how all of these in turn influenced patient depressive symptomatology. RESULTS: Pre diagnosis physical functioning, symptom severity, and days since surgery were significant predictors of physical functioning deficit. Patients who had been treated only with surgery experienced greater physical functioning deficits than did patients who had received both surgery and adjuvant therapy. This apparent anomaly was partly explained by the time interval from surgery to interview. Higher levels of symptom severity, lower levels of prior physical functioning, and greater physical functioning deficits all predicted higher levels of depressive symptomatology. CLINICAL IMPLICATIONS: In the care of elderly patients with cancer, it is important for healthcare providers to consider the pre diagnosis levels of physical functioning of patients with cancer to understand and anticipate the physical and psychological consequences of cancer and its treatment. Equally important is the proper management of patient symptoms in maximizing both the physical and psychological quality of life. PMID- 11879269 TI - Effect of group therapy for breast cancer on healthcare utilization. AB - PURPOSE: The purpose of this study was to determine whether participation in a group psychosocial intervention by patients with breast cancer would result in an improvement in psychological measures and in reduced billings in general medical expenses. DESCRIPTION OF STUDY: Eligible women who had completed treatment for stage 0, I, or II primary breast cancer were prospectively and randomly assigned to either the intervention (n=46) or control (n=43) group. Both groups received the usual psychosocial care; however, the intervention group also participated in six weekly cognitive/behavioral psychosocial meetings. All were assessed on psychiatric symptoms, mood, depression, and coping strategies at four time periods: pre-intervention, post-intervention, 1-year follow-up, and 2-year follow up. Alberta Healthcare billing records were obtained covering the 2-year follow up period to determine the amount billed per person over the course of the study. RESULTS: Women in the intervention group had less depression, less overall mood disturbance, better overall quality of life, and fewer psychiatric symptoms than those in the control group, beginning immediately post-intervention and remaining so at 2 years post-intervention. Billing in the intervention group was an average of $147 less than in the control group, a 23.5% reduction. CLINICAL IMPLICATIONS: This is the first study to show that a psychosocial intervention can reduce direct healthcare billings in a sample of patients with cancer. Importantly, these findings help to justify the routine availability of such programs in cancer treatment facilities worldwide. PMID- 11879270 TI - Lake Superior Rural Cancer Care Project, part I: an interventional trial. AB - PURPOSE: To date, effective cancer care and control intervention studies have been carried out largely in urban and suburban populations. This study was conducted to test innovative interventions, using experimental designs, to improve the care and outcomes of patients with cancer in rural settings. DESCRIPTION OF STUDY: The Lake Superior Rural Cancer Care Project (LSRCCP) tested an innovative, multimodal, multidisciplinary intervention that involved rural healthcare providers and their healthcare system. An experimental design was used, with the rural community as the unit of randomization. Outcomes were measured at three levels: rural providers' knowledge of cancer management, providers' practice performance, and patient outcomes. This 5-year study was conducted in rural areas of northern Minnesota, Wisconsin, and the western part of the Upper Peninsula of Michigan. RESULTS: Baseline data from the study are provided, and details of the design and methods are presented. The study outcomes are reported in part in "Lake Superior Rural Cancer Care Project Part II" in this issue and will be reported further in future issues. CLINICAL IMPLICATIONS: This article describes the hypotheses, design, and methods of the LSRCCP. The design and methods as well as the results of this study may be useful to cancer researchers and clinicians in rural areas across the United States. PMID- 11879272 TI - Creating comprehensive cancer care: a journey. PMID- 11879271 TI - Lake Superior Rural Cancer Care Project, part II: provider knowledge. AB - PURPOSE: The purpose of this article is to report the main learning outcomes of the Lake Superior Rural Cancer Care Project. DESCRIPTION OF STUDY: The authors designed and tested a multimodal intervention directed at rural providers and their healthcare systems in a large rural area in the north central United States. An experimental design was used to randomize rural providers at the group level. The intervention consisted of providing increased education for rural providers with a number of approaches, including the use of clinical opinion leaders. The main outcome of the intervention was knowledge scoring on discipline specific cancer management tests. RESULTS: Knowledge scores for providers in the experimental group significantly increased from pretest to post-test: 66 to 79 for physicians (and physician assistants) (P=.02); 58 to 71 for nurses (P=.01); and 54 to 64 for pharmacists (P=.01). At post-test, participating providers in the experimental group performed significantly better on the knowledge tests (P <.01) than those in the control groups. CLINICAL IMPLICATIONS: This study may be the first to test educational interventions to improve rural providers' knowledge about cancer practice using an experimental design. The intervention may possibly change provider practice behaviors and, thus, patient outcomes, data that will be reported in a future issue. Finally, this educational intervention may prove useful for providers in other rural areas. PMID- 11879273 TI - The challenge of cancer in the workplace. PMID- 11879274 TI - New advances in the management of cutaneous T-cell lymphoma. PMID- 11879275 TI - NIH recommends range of adjuvant breast cancer therapies. PMID- 11879276 TI - Sentinel lymph node dissection: a study in protecting human subjects. PMID- 11879277 TI - The challenge of providing culturally competent cancer care. PMID- 11879281 TI - Problem-solving cancer care education for patients and caregivers. AB - PURPOSE: A program evaluation was conducted to explore the potential effects of a 90-minute problem-solving education session for persons with advanced cancer and their families. DESCRIPTION OF PROGRAM: Patients with advanced cancer and their families, who were visiting a tertiary-care outpatient setting, were invited to attend a 90-minute individualized educational session that taught basic problem solving principles using a cognitive-behavioral framework. Pre-education and posteducation data were collected about the confidence of participants in providing care, their feelings about being informed about resources, and their perceptions of their problem-solving ability. RESULTS: At baseline, most participants reported low confidence about their ability to provide cancer care and felt uninformed about community resources, but they viewed themselves as moderate-to-good problem solvers. Forty-two educational sessions were delivered to 49 caregivers and 40 patients. Two months later, participants reported feeling more informed about community resources and achieved higher posteducation scores for problem-solving ability. More caregivers than patients reported that reading The Home Care Guide for Cancer made a great deal of difference in their approach to home care. CLINICAL IMPLICATIONS: Most educational sessions for families affected by cancer focus on delivering information, not on building skills. These findings suggest that a one-on-one educational session that teaches problem solving skills can be successfully delivered in a busy clinic setting. Family caregivers are especially likely to benefit from this program. PMID- 11879282 TI - Monoclonal antibody therapy for lymphoma: an update. AB - PURPOSE: The purpose of this article is to review the historical development and recent advances in the application of monoclonal antibodies for the treatment of lymphoma. OVERVIEW: The history of clinical applications of monoclonal antibodies has been intertwined with that of lymphomas. The first report of a complete remission in 1981 described a patient with follicular lymphoma who was treated with a murine anti-idiotype antibody. Later that decade there appeared additional encouraging reports of radiolabeled monoclonal antibodies, immunotoxins, and other antibodies with antitumor effects against lymphoma and chronic lymphocytic leukemia. Monoclonal antibodies as a treatment of malignancy became reality in late 1997 when the US Food and Drug Administration approved the anti-CD20 chimeric monoclonal antibody rituximab for the treatment of B-cell lymphoma. Since that time an anti-CD25 monoclonal antibody (dacliximab) and an anti-CD25 immunotoxin fusion product (denileukin diftitox) have become clinically available. Several radio- labeled antibodies, including the murine anti-CD20 products (131)I-tositumomab and (90)Y-ibritumomab tiuxetan, are in advanced stages of clinical testing as are other unlabeled monoclonal antibodies with antilymphoma activity. Other antilymphoma immunotoxins that react with CD25, CD19, and CD22 also have shown promise. CLINICAL IMPLICATIONS: The therapeutic arsenal against lymphoma has been significantly changed by the addition of these antibody products that are active as single agents, and are synergistic, additive, or both with other antilymphoma treatments. PMID- 11879283 TI - Breast and cervical cancer screening among Chinese American women. AB - PURPOSE: The purpose of this study was to describe breast and cervical cancer screening knowledge and practices of a representative sample of Chinese American women and to examine the factors associated with screening practices. DESCRIPTION OF STUDY: A random sample of 332 Chinese American women, 40 to 69 years of age, from the Chinatown area of Chicago, Illinois, were interviewed face to face, using both Chinese Mandarin (or Putunghua) and Cantonese versions of the National Health Interview Survey (NHIS) Cancer Control Supplement Questionnaire. Knowledge and use of mammogram, clinical breast examination (CBE), breast self-examination (BSE), and Pap smear test were assessed. RESULTS: The results showed a low level of knowledge of cancer screening tests and low use rates. Multiple logistics regression analysis showed that women with spoken English fluency were more likely to have knowledge and use of CBE, BSE, Pap test, and mammograms. Women with better than an elementary education were more likely to have knowledge and use of CBE, BSE, and Pap test. The source of medical care was statistically significant for having had a mammogram. Knowledge of cancer warning signs and symptoms was significant for knowledge of mammogram and BSE and for the use of BSE. CLINICAL IMPLICATIONS: Multiple strategies are needed. These might include the following: 1) integration of research with population-based care by physicians and scientists; 2) coordinated public health education on cancer screening and postscreening support in Chinese languages; and 3) replication of the NHIS survey methodology and adaptive application of these instruments across several states and regions, combined with the assessment of screening performance in Chinese American populations. PMID- 11879285 TI - Computed tomography screening for lung cancer: what should we recommend? PMID- 11879284 TI - Educating patients on sentinel lymph node dissection for breast cancer. AB - PURPOSE: This article reviews the current literature on lymphatic mapping and sentinel lymph node dissection (SLND) for breast cancer and presents educational information for patients who are considering undergoing this procedure. OVERVIEW: Lymphatic mapping with SLND has been tested widely in patients with breast cancer, primarily in the context of clinical trials. Research studies have found a high degree of accuracy, with the sentinel lymph node (SLN) predicting the status of the axillary node basin. The ability of the surgeon to identify the SLN and the accuracy of the technique correlate with the number of procedures conducted. With the increase in the number of patients having lymphatic mapping and SLND for breast cancer who may not be part of a clinical trial, there is a need for educational materials to help clinicians teach patients about the procedure. CLINICAL IMPLICATIONS: Because of the complexity of the information, patients need both written and verbal information to decide whether to undergo an SLND. In the setting of a clinical trial, patient education materials add to the informed consent document. As the use of SLND for breast cancer becomes more common, the need for clear, concise, informative patient education materials is even more imperative. PMID- 11879286 TI - Providing resources for Asian Americans and Pacific Islanders. PMID- 11879289 TI - Obesity linked to cancer risk. PMID- 11879287 TI - Epirubicin as adjuvant therapy for breast cancer. PMID- 11879290 TI - Challenging conventional wisdom in cancer care. PMID- 11879291 TI - A patient with renal cell carcinoma, myoclonus, and unrelieved pain. PMID- 11879296 TI - Fatigue and quality of life outcomes of exercise during cancer treatment. AB - PURPOSE: Despite the recognition of fatigue as a common and distressing symptom during cancer treatment, there are few evidence-based interventions available to manage such fatigue. The purpose of this multi-institutional pilot study was to explore the effects of a home-based moderate walking exercise intervention on fatigue, physical functioning, emotional distress, and quality of life (QOL) during breast cancer treatment. DESCRIPTION OF STUDY: Fifty-two women were recruited from five university hospital outpatient departments for this pilot study with an experimental design. Subjects were randomly assigned to the walking program or to usual care during adjuvant chemotherapy or radiation therapy for breast cancer. Symptoms, physical functioning, and QOL were measured at baseline, midtreatment, and at the end of treatment. RESULTS: Women who exercised at least 90 minutes per week on 3 or more days reported significantly less fatigue and emotional distress as well as higher functional ability and QOL than women who were less active during treatment. CLINICAL IMPLICATIONS: A home-based walking exercise program is a potentially effective, low-cost, and safe intervention to manage fatigue and to improve QOL during adjuvant chemotherapy or radiation therapy for breast cancer. This health-promoting self-care activity needs further testing in large randomized clinical trials. PMID- 11879298 TI - Cancer screening among Korean Americans. AB - PURPOSE: Cancer is a leading cause of death among Korean Americans (KAs), one of the most rapidly growing ethnic minority communities in the United States. An understanding about cancer screening practices among Koreans (Ks) living in Korea can be useful in designing culturally sensitive health promotion programs in the United States. The purpose of this study was to describe the cancer screening knowledge and behaviors of a sample of KAs in Los Angeles, California, compared with a similar sample of Ks in Korea. DESCRIPTION OF STUDY: This descriptive survey used a convenience sample of 140 KAs age 40 and older, living in Los Angeles, California, in 1998. A random sample of 149 Ks from Pusan, Korea, in 1995 to 1996 was used as a comparison group. KAs had lived in the United States for an average of 15 years. A Korean and English language 58-item self administered questionnaire assessed knowledge of cancer screening tests and personal cancer screening practices. RESULTS: The findings demonstrate that both KAs and Ks had low participation in cancer screening. Breast cancer screening was significantly more likely among KA women than among Ks; screening rates for cervical cancer and gastrointestinal malignancies were not significantly different between the two groups. CLINICAL IMPLICATIONS: These data can be used to assist healthcare professionals in the development of educational tools and strategies for promoting cancer screening programs for KAs. The unique contribution of this study was the opportunity to compare findings from an immigrant Korean population with a similar population in the country of origin. PMID- 11879297 TI - Screening mammography for elderly women. AB - PURPOSE: Breast cancer is one of the most common and deadly cancers among elderly women, but there is little consensus in the literature about appropriate screening guidelines for the elderly. In this report, evidence-based studies addressing the efficacy and effectiveness of mammography screening programs for elderly women are analyzed and interpreted. OVERVIEW: Although there is little direct scientific evidence to support (or to disavow) the benefits of mammography screening for elderly women, a review of the relevant literature supports the benefits of screening mammography for this population. The authors believe that the current evidence supports the efficacy and effectiveness of mammography screening programs for women up to at least age 80 and also suggests that women older than age 80 who are in good general health will also benefit from screening. CLINICAL IMPLICATIONS: Most individuals rely on their healthcare providers' recommendations to aid them in making decisions regarding their health. When healthcare professionals are equipped with evidence-supported guidelines, they can ensure that appropriate recommendations are being made for their patients. Healthcare professionals who treat the elderly should implement the authors' recommendations, supported by current evidence. The lack of direct evidence should not be interpreted as evidence against the use of screening mammography for the elderly but rather as a call for clinical research on preventive healthcare for senior citizens. PMID- 11879299 TI - Oncologists' and naturopaths' nutrition beliefs and practices. AB - PURPOSE: Despite the widespread use of complementary and alternative therapies by persons with cancer, little is known about how the understandings of complementary and alternative practitioners of these therapies compare with those of conventional physicians. The broad purpose of this research was, thus, to explore the beliefs of physicians and complementary and alternative health practitioners, their use of scientific and other types of evidence, and their counseling practices. The specific issue addressed was the beliefs and practices of oncologists and naturopaths regarding the role of diet in breast cancer prevention and treatment. DESCRIPTION OF STUDY: A qualitative research design was used, involving in-depth semistructured interviews with 10 oncologists and 11 naturopaths. Interviews were tape-recorded, transcribed verbatim, and analyzed using qualitative methods. RESULTS: The oncologists believed that there is little evidence of a role for diet in breast cancer prevention and treatment, citing the lack of evidence from randomized controlled trials. They reported that they provide only general advice on healthy eating to patients. The naturopaths believed that diet is strongly implicated in breast cancer development, prevention, and treatment. They justified this belief by using scientific evidence from a variety of types of studies, logic or common sense, and their clinical experiences. Naturopaths reported that they provide patients with specific suggestions for foods to avoid or to emphasize in diets. CLINICAL IMPLICATIONS: The differences in the advice provided by oncologists and naturopaths are associated with different decision-making or knowledge construction systems. Educating patients about these differences will help patients to make more informed healthcare choices. PMID- 11879300 TI - Acupuncture: evidence and implications for cancer supportive care. PMID- 11879301 TI - Locating and evaluating cancer information on the internet. PMID- 11879303 TI - Arsenic trioxide for the treatment of acute promyelocytic leukemia. PMID- 11879305 TI - Forming partnerships, moving pain management forward. PMID- 11879304 TI - Colorectal cancer screening remains underutilized: CDC report. PMID- 11879306 TI - Metastatic pancreatic cancer: meeting palliative care needs. PMID- 11879307 TI - Physician referral patterns to a breast cancer support program. AB - PURPOSE: The purpose of this study was to describe the differences between physicians who did and did not refer their patients with breast cancer to the American Cancer Society's Reach to Recovery program, as well as to determine the factors associated with these physician referral patterns. DESCRIPTION OF STUDY: This descriptive cross-sectional study included 54 physicians who had and 23 physicians who had not referred their patients with breast cancer to the Mile High Unit Reach to Recovery program in 1999. Participating physicians completed a questionnaire about their knowledge, behaviors, and beliefs regarding Reach to Recovery, the characteristics of the patients they referred, and their own demographic and practice characteristics. RESULTS: General surgeons were more likely than all other specialties to refer patients to the Reach to Recovery program. The program is a part of the American Cancer Society. Feedback from patients about their experiences with Reach to Recovery and the proportion of patient time spent caring for patients with breast cancer were all positively associated with referral. CLINICAL IMPLICATIONS: These findings show that those providing support programs, such as Reach to Recovery, need to use targeted strategies to ensure that physicians are aware of the services they provide and that patients relay their experiences back to their referring physician. Because the value of social support programs for women with breast cancer has been established, physicians can become valuable partners in helping more women gain access to these programs. PMID- 11879308 TI - A weekend camp for bereaved siblings. AB - PURPOSE: The purpose of this article is to describe a weekend bereavement camp for children age 6 to 18 years who have lost a brother or sister to cancer. A description of the planning for the camp and the weekend program is included. OVERVIEW: Camp New Horizons is a weekend camp for children age 6 to 18 years who have lost a sibling to cancer. At the camp, the children are divided into age appropriate groups and spend time, through various activities, learning about the grief process to reduce their feelings of isolation, to express grief appropriately, and to move forward in the grief process. Similar programs, as well as the uniqueness of this program, are discussed. CLINICAL IMPLICATIONS: Camp New Horizons has met many of the educational and support needs of bereaved siblings. Networks of support and friendship have formed that allow the children to reach out to one another when needed. Parents and children have increased their communication about the death in their family, thus accepting their feelings and increasing support in the family system. Continued collaboration between centers will create ongoing support for the healthcare professionals and the programs they provide. PMID- 11879309 TI - Perceived cancer causes: use of complementary and alternative therapy. AB - PURPOSE: The objectives of this report are to describe beliefs about general and personal cancer causes among patients with cancer who use and do not use complementary and alternative medicine (CAM), to compare these beliefs with current epidemiologic knowledge, and to explore associations between these beliefs and choice of CAMs. DESCRIPTION OF INTERVIEWS: The authors conducted a semistructured interview with 143 patients with cancer, who were identified via the Hawaii Tumor Registry and had participated in a survey on CAM use. By design, the majority of interviewees (85%) were CAM users. A qualitative data analysis was performed. RESULTS: Genetics, environment, and diet were among the most common perceived general causes of cancer, whereas psychosocial factors were by far the most common perceived personal causes. CAM users were more likely than nonusers to name environment, immune system, and stress as cancer causes. Compared with the epidemiologic literature, study participants were less likely to mention tobacco use and more likely to consider environmental exposures, psychosocial factors, immunologic mechanisms, and physical injury as causes of cancer. Furthermore, the interviews suggest that causal thinking may be related to the type of CAM used. CLINICAL IMPLICATIONS: Given the discrepancy between popular cancer theories and current scientific knowledge, the challenge for the healthcare provider is to listen and try to understand the patient's beliefs about cancer etiology. Understanding the patient's beliefs about their disease is important in providing the support and information they need to make effective decisions about their medical care. PMID- 11879310 TI - Cancer clinical trial patients in the information age: a pilot study. AB - PURPOSE: The purpose of this pilot study was to explore the cancer clinical trial patient's perception and use of electronic information and communication. DESCRIPTION OF STUDY: A total of 42 participants who self-reported they had enrolled, or were enrolled, in a cancer clinical trial completed a Web-based clinical trials survey questionnaire linked to a cancer-specific Web site. The questionnaire was used to obtain written responses to open-ended questions regarding the perception and use of electronic information and communication. RESULTS: The pilot study findings document that more than one third of the respondents use the Internet to seek clinical trials information. Results also describe the impact of the Internet in helping these patients to identify what questions to ask their healthcare providers. The findings underscore the desire of patients to communicate not only with others on the same clinical trial, but also with their healthcare providers via the Internet. CLINICAL IMPLICATIONS: When providing care to cancer clinical trial patients in the information age, online information seeking and the desire for electronic communication must increasingly be acknowledged, understood, and supported. A proactive and comprehensive approach to addressing patient information and support needs can facilitate patient empowerment and satisfaction. PMID- 11879311 TI - Identifying support issues of parents of children with leukemia. AB - PURPOSE: One of the important factors in helping both children with acute lymphoblastic leukemia (ALL) and their family members during the difficult treatment period is the support they receive. However, there is a scarcity of research that examines the adaptation to such an experience from the family's perspective. This discussion presents the findings from recent research on accessing support from the perspective of parents with a child undergoing treatment for ALL. DESCRIPTION OF STUDY: A longitudinal, qualitative study informed by a phenomenological methodology. The results are taken from parent interviews undertaken during the time period from the end of induction-remission to the end of the 5-week period (time period T1), collected over the first year of operation of the study. RESULTS: The need for support was reported as intense, and offers of support tend to lessen over time. Among the sources of support noted were partners, family, friends, employers, hospital staff, and other parents in the same situation. However, community resources appeared to be lacking, underused, or both. CLINICAL IMPLICATIONS: Suggestions for support include the provision of practical assistance through appropriately trained volunteers, the involvement of a liaison person to mentor families through the initial stages of treatment, development of parent support groups, and the opportunity for families to meet children who are successfully off treatment. Such support strategies involve sensitivity to particularly vulnerable families who have limited support available and respect for the variation in individual styles of accessing support. PMID- 11879312 TI - Multimedia resources for pain education. PMID- 11879313 TI - Alemtuzumab: a new option for refractory chronic lymphocytic leukemia? PMID- 11879315 TI - Tobacco is major health threat among U.S. women and girls: Surgeon General. PMID- 11879316 TI - Celebrating progress in treatment, renewing our commitment to prevention. PMID- 11879317 TI - Addressing treatment options in metastatic melanoma. PMID- 11879318 TI - Implementing cancer pain education for medical students. AB - PURPOSE: The purpose of this study was to develop and pilot a cancer pain education course for medical students, using a structured home hospice visit. DESCRIPTION OF STUDY: A 1-hour home hospice visit was presented to 57 senior medical students. The content and objective criteria for the structured home hospice visit were developed by a multidisciplinary group of experts. During a 1 hour interview, students completed a cancer pain history, performed a focused physical examination, and received feedback and teaching regarding the essentials of cancer pain management from the hospice nurse. All students and hospice patients completed a multi-item evaluation questionnaire with a 5-point Likert scale (1=strongly disagree; 5=strongly agree) regarding the structured home hospice visit. RESULTS: Most students agreed strongly that the home hospice visit was a positive experience (mean +/- SD 4.8 +/- 0.44) that helped them to understand the management of cancer pain (mean 4.7 +/- 0.46) and opioid-related side effects (mean 4.5 +/- 0.57). Most patients enjoyed visiting with the students (mean 4.90 +/- 0.30), agreed that the visit was not tiring (mean 4.81 +/ 0.51), and felt that they benefited from participating (mean 4.76 +/- 0.54). CLINICAL IMPLICATIONS: The authors concluded the following: 1) that medical students benefited from learning about cancer pain assessment and management through the use of a structured home hospice visit; 2) that a structured home hospice visit helped the students to learn the basics of cancer pain management; 3) that patients enjoyed their role as teacher for medical students; and 4) that senior hospice nurses provided excellent instruction for medical students in the management of cancer pain. PMID- 11879319 TI - Feasibility of assessing quality in medical oncology practices. AB - PURPOSE: This study was designed to test the feasibility of conducting routine quality assessment within community medical oncology practices. DESCRIPTION OF STUDY: Eleven medical oncologists practicing within the Fox Chase Network were surveyed over an 8-month period, using a standardized 10-item checklist. Eight of the items (ie, board certification, continuing education, office procedure manual for chemotherapy, chemotherapy flow sheets, body surface area calculations, adherence to guidelines for follow-up of breast cancer, adjuvant hormones in women with estrogen receptor-positive breast cancer, and patient satisfaction survey) were chosen because they measure structural and process variables particularly relevant to the high-volume clinical services seen in private practice oncology. The authors also calculated two rates (protocol accrual and neutropenic complications of chemotherapy) to test as putative indicators of quality. RESULTS: The authors found a high level of both physician interest in developing the audit measures and compliance with the survey process. Overall quality of care, as measured by structure and process variables, was excellent with negligible internal variability. Derived rates of protocol accrual (0.003 0.373; mean 0.11, SD 0.11) and neutropenic sepsis (0.004-0.014; mean 0.007, SD 0.004) show considerable variability, however, and are only minimally correlated (r= -.36). These are both potential indicators of quality that should be further evaluated. CLINICAL IMPLICATIONS: The authors have demonstrated the feasibility of conducting quality assessment within private medical oncology practices and have identified two easily calculated rates that merit further study as potential indicators of quality. PMID- 11879320 TI - Fears about breast cancer recurrence:. AB - PURPOSE: Prior research shows that the majority of women diagnosed with breast cancer experience ongoing fears about recurrence. These studies primarily have used Likert scales to determine the extent to which women fear recurrence but have not presented details of women's experiences or addressed the nature of women's fears. This report describes a pilot study designed to capture women's diverse experiences related to the possibility of recurrence. DESCRIPTION OF STUDY: Sixteen women participated in semi-structured interviews about their thoughts and feelings regarding the possibility of breast cancer recurrence. Thematic content coding was used to categorize women's responses. RESULTS: Although some women indicated strong fears about recurrence, the majority expressed more moderate fears. Fears revolved around the possibility of death, further treatment (particularly chemotherapy), emotional difficulties, pain, advancement of the disease, suffering of family members, and loss of the breast. Many women spoke about life circumstances that were more salient than the possibility of recurrence. Women also described events/situations that triggered fears and strategies used for coping with fears. CLINICAL IMPLICATIONS: Results suggest that more systematic investigation of women's fears about breast cancer recurrence is warranted and that clinicians should address the multidimensional nature of women's fears as they relate to other psychological processes and events in women's lives. Moreover, women's life circumstances should be considered when attempting to determine whether a social support intervention might be beneficial or detrimental. PMID- 11879321 TI - Altered sexual patterns after treatment for prostate cancer. AB - PURPOSE: The purpose of this study was to identify and describe the impact of prostate cancer and its treatment on men's sexuality and intimate relationships from the perspective of the men's lived experiences. DESCRIPTION OF THE STUDY: Ten men diagnosed with prostate cancer were interviewed. Verbal transcripts were analyzed using an interpretive phenomenological approach based on Heidegger's philosophy. RESULTS: "Altered sexual patterns" was identified as the major theme, which in turn was influenced by five minor themes: choice of consequences; age affecting sexual life; hope of improvement; acceptance of altered sexual life; and image of manliness. Choice of consequences showed that survival is paramount, as interviewees often felt that they were choosing between death or sexual dysfunction. Age affecting sexual life identified sexual dysfunction as caused by natural effects of aging. Hope of improvement showed that several men hoped for improvement in sexual function, although it had been years since their treatment. Acceptance of altered sexual life illustrated that the men often felt the need to accept their altered sex life, with their wives playing an important role in this acceptance. Finally, the theme image of manliness showed the men's attempts to acknowledge and cope with the changes in their self-image. CLINICAL IMPLICATIONS: It is important that healthcare professionals adopt a comprehensive approach in communicating information about prostate cancer treatment choices and potential side effects that includes the patient and his partner. Communication and counselling about treatment options, potential side effects, and potential strategies to manage side effects should be offered both at the time of diagnosis and at post-treatment follow-up. PMID- 11879322 TI - Select complementary/alternative therapies for prostate cancer: the benefits and risks. PMID- 11879323 TI - National Cancer Institute resources for patients and their caregivers. PMID- 11879324 TI - Imatinib mesylate: a new pill for chronic myelogenous leukemia. PMID- 11879326 TI - CDC report on 20 years of AIDS: more than one million americans infected. PMID- 11879327 TI - Embryonic stem cell research: balancing promise and peril. PMID- 11879328 TI - Communication challenges in a young man with Hodgkin's disease. PMID- 11879329 TI - Barriers to fecal occult blood testing and sigmoidoscopy among older Chinese American women. AB - PURPOSE: This study examined factors associated with fecal occult blood test (FOBT) and sigmoidoscopy screening use among Chinese-American women age 60 years and older. DESCRIPTION OF STUDY: One hundred women were recruited from senior centers in two metropolitan areas on the east coast of the United States. Participants completed a questionnaire that included sections on demographics, health history, health insurance coverage, FOBT and sigmoidoscopy use, common and cultural barriers to colorectal cancer screening, and acculturation. RESULTS: Logistic regression models found greater acculturation to be a significant predictor of having had a FOBT at least once, and found both greater acculturation and physician recommendation to be significant predictors of having had a sigmoidoscopy at least once. No significant predictors were found for regular adherence to colorectal screening guidelines, which include having undergone an FOBT in the past year and sigmoidoscopy in the past 5 years. CLINICAL IMPLICATIONS: This study found that older Chinese-American women underuse FOBT and sigmoidoscopy screening, as is recommended by the American Cancer Society colorectal cancer screening guidelines. These findings suggest that cultural factors may influence the initiation of colorectal cancer screening for Chinese-American women but are not predictive of adherence to screening over time. Outreach efforts to promote colorectal cancer screening in this population might target women who are less acculturated to facilitate an initial entry into the Western healthcare system to obtain screening. PMID- 11879330 TI - Psychological sequelae and alopecia among women with cancer. AB - PURPOSE: This article reviews the relevant literature on treatment-induced alopecia in women with cancer and describes the development of a computer assisted intervention to reduce distress associated with this side effect. DESCRIPTION OF PROGRAM: Alopecia has been cited as the most disturbing anticipated side effect by up to 58% of women preparing for chemotherapy, with 8% being at risk for avoiding treatment. Women with cancer who experience alopecia as a side effect, compared with women with cancer and no alopecia, report lower self-esteem, poorer body image, and lower quality of life. Although physicians' recommendations are the most influential factor on cancer treatment choice, body image and effects on sexuality are the next most influential factors. A study of a computer-imaging intervention, based on concepts related to guided imagery and anticipatory grief, has been launched in an effort to aid women in coping with anticipated treatment-related alopecia. RESULTS: While we are still waiting for final data collection and analysis from the computer intervention study, the feedback thus far has been positive. CLINICAL IMPLICATIONS: The intervention described here may prove to be effective in desensitizing women with cancer to hair loss and facilitating an adjustment to self-acceptance. As such, a higher quality of life during the difficult time of coping may be maintained. The development of a computer-imaging intervention offers an opportunity to integrate a standard psychosocial intervention, personalized for each patient, into the routine patient care in the oncology setting. PMID- 11879331 TI - Cancer and folie a deux: case report, treatment, and implications. AB - PURPOSE: This report describes the uncommon psychopathological state best known as folie a deux, or shared psychotic disorder, in a unique case in which the grandiose, religious delusions of a woman with uterine cancer are shared with her husband. OVERVIEW: More than 50% of patients with cancer meet the criteria for diagnosis of major psychiatric disorders. Certainly, these disorders may occur as a result of the stress of the cancer diagnosis and treatment, but also because of a predisposition to psychiatric illness or a pre-existing psychiatric illness. In the medical setting, the phenomenon of folie a deux poses significant problems not only for the patient, but also for the involved family member, psychiatric consultant, and healthcare team. CLINICAL IMPLICATIONS: In this case, the patient's delusional system had a grave impact on her ability to make rational healthcare decisions, for which she was deemed incompetent. The first choice for her healthcare surrogate, her husband, was so affected by his sharing of her psychotic condition that he could not fulfill this role. In assessing a suspected case of folie a deux, awareness of several issues--the point at which religious over-ideation becomes delusional, the spectrum of competency, informed consent, and treatment refusal--is important. PMID- 11879332 TI - The treatment of prostate cancer: an overview of current options. AB - PURPOSE: The purpose of this report is to discuss the current treatment options available to the patient with prostate cancer in all stages of the disease. OVERVIEW: With the exception of skin cancer, prostate cancer is the most common cancer in men in the United States. Most patients in the current era will present with organ-confined disease, amenable to curative treatment. Treatment for organ confined disease includes watchful waiting, radical prostatectomy, radiation therapy, and cryosurgery in selective cases. Hormone therapy is the cornerstone of treatment of patients with advanced prostate cancer. There is no curative treatment for hormone-refractory prostate cancer. CLINICAL IMPLICATIONS: The availability of several therapeutic options for localized prostate cancer warrants careful consideration when planning treatment with curative intent. Patients need to be active participants in decision making, and they must be aware of the benefits and possible complications of the different types of treatment. Patients with advanced prostate cancer need to be aware that hormone treatment will provide temporization and palliation in the majority of cases. Hormone-resistant prostate cancer is refractory to most forms of conventional and experimental therapy. PMID- 11879333 TI - Complementary and alternative therapy for breast cancer: the evidence so far. PMID- 11879334 TI - Patient resources for cancer-related fatigue. PMID- 11879335 TI - Caspofungin: the first of a new class of antifungal agents. PMID- 11879337 TI - Caloric restriction inhibits seizure susceptibility in epileptic EL mice by reducing blood glucose. AB - PURPOSE: Caloric restriction (CR) involves underfeeding and has long been recognized as a dietary therapy that improves health and increases longevity. In contrast to severe fasting or starvation, CR reduces total food intake without causing nutritional deficiencies. Although fasting has been recognized as an effective antiseizure therapy since the time of the ancient Greeks, the mechanism by which fasting inhibits seizures remains obscure. The influence of CR on seizure susceptibility was investigated at both juvenile (30 days) and adult (70 days) ages in the EL mouse, a genetic model of multifactorial idiopathic epilepsy. METHODS: The juvenile EL mice were separated into two groups and fed standard lab chow either ad libitum (control, n=18) or with a 15% CR diet (treated, n=17). The adult EL mice were separated into three groups; control (n=15), 15% CR (n=6), and 30% CR (n=3). Body weights, seizure susceptibility, and the levels of blood glucose and ketones (beta-hydroxybutyrate) were measured over a 10-week treatment period. Simple linear regression and multiple logistic regression were used to analyze the relations among seizures, glucose, and ketones. RESULTS: CR delayed the onset and reduced the incidence of seizures at both juvenile and adult ages and was devoid of adverse side effects. Furthermore, mild CR (15%) had a greater antiepileptogenic effect than the well-established high-fat ketogenic diet in the juvenile mice. The CR-induced changes in blood glucose levels were predictive of both blood ketone levels and seizure susceptibility. CONCLUSIONS: We propose that CR may reduce seizure susceptibility in EL mice by reducing brain glycolytic energy. Our preclinical findings suggest that CR may be an effective antiseizure dietary therapy for human seizure disorders. PMID- 11879338 TI - Contrasting effects of zonisamide and acetazolamide on amygdaloid kindling in rats. AB - PURPOSE: Zonisamide (ZNS) and acetazolamide (AZM) are two antiepileptic drugs (AEDs) that differ in clinical efficacy. To elucidate the mechanisms of action of these compounds, we investigated their therapeutic and prophylactic effects in rats by using a kindling model of partial epilepsy. METHODS: Electrodes were implanted into the left amygdala of adult male Wistar rats. The animals were stimulated at the afterdischarge threshold until five stage 5 seizures were induced. The generalized seizure threshold was then determined. Therapeutic effects were examined in rats manifesting successive convulsions with near threshold stimulation. To test prophylactic effects, drugs were administered intraperitoneally before daily kindling stimulation until the animal had a stage 5 seizure or reached day 18. RESULTS: ZNS (10-40 mg/kg; n=6) suppressed kindled seizures in a dose-dependent manner. Repeated administration for 7 days produced tolerance to anticonvulsive effects. AZM (25-200 mg/kg; n=7) showed limited therapeutic effect, alleviating only the clonic convulsion in stage 5 seizures and reducing afterdischarge duration. Secondary generalization was not significantly suppressed during repeated treatment (50-200 mg/kg; n=6). ZNS, 25 or 40 mg/kg (n=8), significantly retarded seizure development; 15.0 or 17.0 daily stimulations were required to produce a stage 5 seizure. AZM, 50-200 mg/kg (n=6), also retarded seizure development, with 14.0-14.8 stimulations required. CONCLUSIONS: ZNS exhibited modest therapeutic and prophylactic effects, whereas AZM showed mainly prophylactic effects. Hypotheses are presented that may explain the mechanisms of action of these drugs. PMID- 11879340 TI - Adult-onset idiopathic generalized epilepsy: clinical and behavioral features. AB - PURPOSE: To identify and define clinical and behavioral features of patients with adult-onset idiopathic generalized epilepsy (IGE). METHODS: We reviewed the charts of 313 IGE patients at the NYU Comprehensive Epilepsy Center over the past 5 years to identify patients with adult onset (18 years old or older). We excluded patients with childhood or adolescent symptoms that suggested absence, myoclonic, or tonic-clonic seizures, as well as those with a history of significant head injury or other known causes of localization-related epilepsy. RESULTS: Forty-two (13.4%) patients had a clear onset of IGE in adulthood; average age of onset was early 20s (mean, 23.8 years; range, 18-55 years). Twenty one patients had adult myoclonic epilepsy (AME, 50%), and three had generalized tonic-clonic seizures on awakening (GTCS-A, 7%). More than two thirds (n=30) are well controlled with current antiepileptic drugs (AEDs), and almost 90% are currently employed (n=37). One third were diagnosed and treated for mental disorders, including depression (n=12), anxiety (n=7), obsessive-compulsive personality disorder (n=2), and postictal psychosis (n=1). CONCLUSIONS: Adult onset IGE is associated with a good prognosis. An association may exist between psychological disorders, psychotropic medication, and level of seizure control in adults with IGE. PMID- 11879339 TI - Monotherapy versus polytherapy for epilepsy: a multicenter double-blind randomized study. AB - PURPOSE: Monotherapy has been the gold standard in epilepsy treatment for the last 20 years, partly because of the reputation for increased toxicity of polytherapy. However, monotherapy and polytherapy have not been compared in a double-blind clinical trial. Open trials that compared the two treatments were not optimally designed and compared the two at unequal drug loads (i.e., at nonequivalent dosages). We report on a double-blind clinical trial in which a combination of carbamazepine (CBZ) and valproate (VPA) was compared with CBZ monotherapy. Patients started with equal drug loads, and neurotoxicity was the primary outcome measure. METHODS: The 130 adult patients with untreated generalized tonic-clonic and/or partial seizures were randomized to equal drug loads of either monotherapy (400 mg CBZ per day) or polytherapy (200 mg CBZ plus 300 mg VPA per day). Outcome was measured by seizure counts, clinimetric epilepsy scales, and neuropsychological tests at baseline, at 2 and 12 months, and irregularly between 2 and 12 months. RESULTS: No statistical differences were found between the two treatments in the reduction of seizure frequencies, in overall neurotoxicity, or in overall systemic toxicity. The frequencies and clinimetric scores of certain adverse effects did differ (e.g., more monotherapy patients remained sedated, and more polytherapy patients gained weight). Fewer polytherapy patients withdrew because of adverse effects (14 vs. 22%), although this did not reach statistical significance (p=0.15). Neuropsychological assessment did not show significant differences. CONCLUSIONS: No differences were found in overall neurotoxicity between monotherapy and polytherapy. PMID- 11879341 TI - Idiopathic generalized epilepsies: do sporadic and familial cases differ? AB - PURPOSE: Genetic factors are the only identified cause of idiopathic generalized epilepsies (IGEs), but the majority of cases do not have affected first-degree relatives. Here we investigate whether subjects with sporadic and familial IGE differ in terms of antecedent events and clinical and EEG features. Differences would support the hypothesis of a different etiology for sporadic cases, which has implications for choice of subjects for genetic association studies. METHODS: We analyzed 98 patients with IGE, diagnosed on clinical and EEG criteria. All patients and, if possible, one relative were interviewed, with special emphasis on potential antecedent events and family history. Patients with first-degree relatives affected with epileptic seizures were regarded as "familial," and the other patients were regarded as "sporadic." RESULTS: Of the 98 IGE patients, 32 (33%) patients were familial. The risk for seizures was 13.2% for siblings, and 7.7% for parents. The distribution of the IGE subsyndromes, the presence of antecedent events, and other electroclinical features did not differ between familial and sporadic IGE groups. CONCLUSIONS: No differences were found between familial and sporadic IGE patients. This does not the support the hypothesis that sporadic and familial IGE cases have separate etiologies. PMID- 11879342 TI - Relationships among hippocampal volumetry, proton magnetic resonance spectroscopy, and verbal memory in temporal lobe epilepsy. AB - PURPOSE: To examine the relationship between hippocampal volumes, 1H magnetic resonance spectroscopy (MRS)-identified hippocampal metabolic function, and verbal memory in patients with unilateral mesial temporal lobe epilepsy (MTLE). METHODS: Hippocampal volumes, 1H MRS-derived hippocampal creatine to N acetylaspartate (Cr/NAA), and verbal memory assessment were obtained preoperatively in 22 patients (six right, 16 left) with EEG-defined unilateral MTLE. RESULTS: Left hippocampal volume correlated significantly with left hippocampal Cr/NAA (r=-0.549, p < 0.01), whereas right volume correlated significantly with right Cr/NAA (r=-0.478, p < 0.05). Verbal memory correlated significantly with left hippocampal Cr/NAA (r=-0.594, p < 0.01), but not with left hippocampal volume or right hippocampal measures. CONCLUSIONS: Hippocampal volumes and 1H MRS-derived metabolite ratios are statistically related, but share only a small percentage of variance, suggesting separate but related pathophysiologic processes. Left hippocampal Cr/NAA appears to be more sensitive to verbal memory function than volumes. PMID- 11879343 TI - Memory outcome after left anterior temporal lobectomy in patients with expected and reversed Wada memory asymmetry scores. AB - PURPOSE: The ideal candidate for anterior temporal lobectomy surgery shows a Wada memory asymmetry (WMA) score characterized by better memory performance in the hemisphere contralateral to the seizure focus relative to the ipsilateral (surgical) hemisphere. However, some surgical candidates show a reversed WMA or better Wada memory performance in the hemisphere of surgical interest relative to the hemisphere contralateral to the seizure focus. To date, no data are available contrasting memory and seizure outcome for these two Wada groups. The present study compared memory and seizure outcome after left anterior temporal lobectomy (L-ATL) in patients showing expected and reversed WMA scores, and also examined the relationship of the individual hemisphere Wada memory scores for predicting verbal memory outcome after L-ATL. METHODS: We compared 6-month postoperative verbal memory change scores and seizure outcome in L-ATL patients with either an expected (n=12) or reversed WMA (n=9) pattern on Wada memory testing. RESULTS: L ATL patients showing a reversed WMA score had a poorer verbal memory outcome and poorer seizure control after surgery compared with patients showing a WMA score in the expected direction. CONCLUSIONS: L-ATL patients with a reversed WMA score have a greater risk for memory morbidity and poorer seizure outcome than do patients with a WMA score in the expected direction. The WMA score was the best predictor of memory outcome after L-ATL. When the WMA score is not considered, both individual Wada hemisphere scores (contralateral and ipsilateral) provided significant and independent contribution to predicting postoperative verbal memory functioning. These findings are discussed in the context of the functional reserve and hippocampal adequacy models of memory change after temporal lobectomy. PMID- 11879344 TI - Long-term seizure outcome and antiepileptic drug treatment in surgically treated temporal lobe epilepsy patients: a controlled study. AB - PURPOSE: To evaluate the long-term impact of surgical treatment on seizure outcome and antiepileptic drug (AED) use in patients with pharmacoresistant temporal lobe epilepsy (TLE). METHODS: Comparison of seizure outcome and AED us in operated-on TLE patients (n=148) and nonsurgically treated TLE patients (n=94) at a baseline visit and a follow-up visit after a mean period of 4.8 years. RESULTS: At follow-up, 44.6% of the surgical patients and 4.3% of the nonsurgical patients had been continuously seizure- free since the baseline visit (including the immediate postoperative period). A further 17.6% of the operated-on and 3.2% of the not operated-on patients had been seizure-free for at least the previous year; 37.8% of the surgical and 92.5% of the nonsurgical patients had had seizures during the previous 12 months (p < 0.001). Of the surgical patients, 8.8% versus none of the nonsurgical patients were AED free at follow-up; 55.4% versus 20.2% were receiving monotherapy, and 35.8% versus 79.8% were receiving polytherapy (p < 0.001). Mean number of AEDs and mean change in number of AEDs were significantly more favorable in operated-on than in non-operated-on patients. Further subgroup analysis revealed that not only the continuously seizure-free surgical patients, but also the operated-on patients with ongoing seizures took fewer AEDs than their respective non-operated-on counterparts. CONCLUSIONS: This controlled study for the first time provides comprehensive information on long-term seizure outcome and AED use in surgical TLE patients. It shows a more favorable seizure outcome and AED use in the surgically treated patients. The latter holds true even for the not seizure-free patient subgroup. PMID- 11879345 TI - Pregnancy registries in epilepsy. AB - The risk of major malformations in the offspring of mothers with epilepsy receiving antiepileptic drugs is 4--8% compared to 2--4% in the general population. Risk factors include daily dose and polytherapy. Selected drugs have been found to be associated with a higher risk of specific malformations (congenital heart defects and cleft palate with phenytoin and barbiturates; neural tube defects with valproate and carbamazepine). Although some of these findings are unquestionable, several questions are still unsolved, depending the characteristics of the target populations, the small samples of patients, and the design and limiting factors of the published reports. In the last decade, pregnancy registries have been activated by collaborative groups of physicians in Europe (EURAP), North America (NAREP), Australia and India (the latter two recently merged into EURAP), to enroll a large number of exposed women to be monitored prospectively with standardized methods, and by three pharmaceutical companies marketing lamotrigine, gabapentin and vigabatrin, as part of their post marketing surveillance. Even though the structure of these registries and the target populations should theoretically result in the identification of a sufficient number of women exposed to different drugs and examined for the occurrence of malformations of any type and severity, the implementation of a common database with information from the existing registries may provide valuable information in a shorter time period. Although differences between some of the registries limit the possibility to pool data, a gradual development of a collaboration is highly desirable to discuss a list of design issues and assess to what extent and how data could be compared and organized. PMID- 11879346 TI - Lateralizing and localizing values of ictal onset recorded on the scalp: evidence from simultaneous recordings with intracranial foramen ovale electrodes. AB - PURPOSE: The value of scalp recordings to localize and lateralize seizure onset in temporal lobe epilepsy has been assessed by comparing simultaneous scalp and intracranial foramen ovale (FO) recordings during presurgical assessment. The sensitivity of scalp recordings for detecting mesial temporal ictal onset has been compared with a "gold standard" provided by simultaneous deep intracranial FO recordings from the mesial aspect of the temporal lobe. As FO electrodes are introduced via anatomic holes, they provide a unique opportunity to record simultaneously from scalp and mesial temporal structures without disrupting the conducting properties of the brain coverings by burr holes and wounds, which can otherwise make simultaneous scalp and intracranial recordings unrepresentative of the habitual EEG. METHODS: Simultaneous FO and scalp recordings from 314 seizures have been studied in 110 patients under telemetric presurgical assessment for temporal lobe epilepsy. Seizure onset was identified on scalp records while blind to recordings from FO electrodes and vice versa. RESULTS: Bilateral onset (symmetric or asymmetric) was more commonly found in scalp than in FO recordings. The contrary was true for unilateral seizure onset. In seizures with bilateral asymmetric onset on the scalp, the topography of largest-amplitude scalp changes at onset does not have localizing or lateralizing value. However, 75-76% of seizures showing unilateral scalp onset with largest amplitude at T1/T2 or T3/T4 had mesial temporal onset. This proportion dropped to 42% among all seizures with a unilateral scalp onset at other locations. Of those seizures with unilateral onset on the scalp at T1/T2, 65.2% showed an ipsilateral mesial temporal onset, and 10.9% had scalp onset incorrectly lateralized with respect to the mesial temporal onset seen on FO recordings. In seizures with a unilateral onset on the scalp at electrodes other than T1/T2, the proportions of seizures with correctly and incorrectly lateralized mesial temporal onset were 37.5 and 4.2%, respectively. Thus the ratio between incorrectly and correctly lateralized mesial temporal onsets is largely similar for seizures with unilateral scalp onset at T1/T2 (16.7%) and for seizures with unilateral scalp onset at electrodes other than T1/T2 (11.2%). The onset of scalp changes before the onset of clinical manifestations is not associated with a lower proportion of seizures with bilateral onset on the scalp, or with a higher percentage of mesial temporal seizures or of mesial temporal seizures starting ipsilateral to the side of scalp onset. In contrast, the majority (78.4%) of mesial temporal seizures showed clinical manifestations starting after ictal onset on FO recordings. CONCLUSIONS: A bilateral scalp onset (symmetric or asymmetric) is compatible with a mesial temporal onset, and should not deter further surgical assessment. Although a unilateral scalp onset at T1/T2 or T3/T4 is associated with a higher probability of mesial temporal onset, a unilateral onset at other scalp electrodes does not exclude mesial temporal onset. A unilateral scalp onset at electrodes other than T1/T2 is less likely to be associated with mesial temporal onset, but its lateralizing value is similar to that of unilateral scalp onset at T1/T2. The presence of clinical manifestations preceding scalp onset does not reduce the localizing or lateralizing values of scalp recordings. PMID- 11879347 TI - Childhood epilepsy due to neurocysticercosis: a comparative study. AB - PURPOSE: To assess the clinical profile of pediatric patients with epilepsy and neurocysticercosis (NC), and compare them with a group of pediatric patients with benign partial epilepsy to determine clinical differences, response to treatment, and prognosis. METHODS: We studied 28 patients (16 girls) with probable or definitive diagnosis of NC and epilepsy and 32 patients (16 girls) with partial benign epilepsy (BE). All patients had normal neurologic examination. We compared NC and BE patients looking for differences in demographics (age at first seizure, gender, family history); clinical presentation (type, frequency, duration, and total number of seizures, duration of epilepsy, status epilepticus, cluster, and postictal deficit); treatment [duration, number of antiepileptic drugs (AEDs), maximal dose, drug association, number of seizure-free patients, time to obtain control and recurrence after medication discontinuation]; complementary examinations (the first and the last EEG). RESULTS: The mean follow-up was 5.4 years for the 28 NC patients and 4.6 years for the 32 BE patients (p=0.98). We did not find statistical differences between NC and BE in gender, family history, types of seizures, frequency and length of seizures, previous status epilepticus, seizure clustering, and presence of postictal deficits. However, we found that NC compared with BE patients had significant longer AED treatment, more seizures after AED introduction, tried more AEDs and at maximal dose, and in 20%, required polytherapy. The recurrence rate in NC was 54.4% and this was not significantly associated with number of lesions and disease activity seen on CT scans or the presence of EEG abnormalities. CONCLUSIONS: NC presents with a mild form of epilepsy in terms of seizure severity; however, it is more challenging in regard to drug management and has a less favorable long-term prognosis in terms of seizure remission. The number of lesions or disease activity seen on computed tomography (CT) as well as EEG abnormalities have no prognostic value in childhood epilepsy due to NC. PMID- 11879348 TI - Carnitine levels and the ketogenic diet. AB - PURPOSE: To determine the long-term effect of the ketogenic diet (KD) on carnitine levels and whether carnitine depletion is a significant cause of clinical complications during KD initiation or treatment. METHODS: Carnitine levels at 0, 1, 6, 12, and 24 months of diet treatment, carnitine antiepileptic drug (AED) history, lowest blood glucose and time to achieve ketosis during diet initiation, and diet complications were analyzed for 38 consecutive patients who initiated the KD from May 1997 to March 2000. Carnitine levels at follow-up were analyzed for eight patients started on the diet before to May 1997. RESULTS: Total carnitine (TC) at diet initiation correlated negatively with the number of AEDs at diet initiation but not with number of past AEDs, lowest blood glucose, or time to ketosis. TC decreased in the first months of diet treatment and then stabilized or increased slightly with long term treatment. Only 19% of patients were supplemented with carnitine for low TC. No patient showed clinical signs of carnitine deficiency. CONCLUSIONS: Multiple AED exposure lowers TC, but actual TC deficiency in patients initiating the KD is not common, and TC levels do not appear to predict hypoglycemia or problems achieving ketosis. Mild carnitine depletion may occur early in KD treatment and occasionally TC decreases out of the normal range, without clinical symptoms. TC stabilizes or increases back toward baseline with long-term treatment, and most patients do not require carnitine supplementation. PMID- 11879349 TI - Parents' perceptions of adversity introduced by upheaval and uncertainty at the onset of childhood epilepsy. AB - PURPOSE: We report the parent's perceptions of and reactions to the onset of "epilepsy only" and the implications for continuity of parenting. METHODS: Content analysis was used to extract data on perceived (dis)continuity of parenting, from interviews held with parents of 69 schoolchildren in whom idiopathic or cryptogenic epilepsy ("epilepsy only") had recently been diagnosed. RESULTS: Almost half of the parents (42%) perceived neither themselves nor their child as having been thrown off balance by the onset of epilepsy. Quite a few parents (33%) perceived themselves rather than their child as having been thrown off balance. More parents of children with cryptogenic than with idiopathic epilepsy perceived themselves as being off balance. However, parents' perceptions of their children's reactions to the epilepsy-related changes were not influenced by any epilepsy variable. Rather, family trouble, long-standing behavioral problems, and adolescence contributed to the child's maladaptive reaction. Three extremely off-balance parents had children with seizures of "unclassifiable" epilepsy that later were found to be nonepileptic and psychogenic. CONCLUSIONS: The majority of the parents perceived their child as adapting well to the onset of epilepsy only. Children with seizure onset in adolescence and children with other adversities were perceived as adapting poorly to the additional adversity of epilepsy. Cryptogenic rather than idiopathic etiology leaves parents in great suspense. In behavioral studies, it is advisable to treat children with unclassifiable epilepsy as a separate group. PMID- 11879350 TI - Outcome of severe refractory status epilepticus in children. AB - PURPOSE: Refractory status epilepticus (RSE) is the persistence of seizure activity despite appropriate therapy; it is treated with high-dose suppressive anesthetic agents. We report here the outcome of RSE in a large series of children. METHODS: We retrospectively reviewed cases of RSE treated at Children's Hospital, Boston, between 1992 and 2000. Factors evaluated included age, history of seizures or neurologic impairment, etiology, outcome, including mortality or return to baseline, and initial EEG findings. RESULTS: Twenty-two patients ages 4.5 months to 18 years were admitted to the intensive care unit for RSE. All were treated with high-dose suppressive therapy consisting of pentobarbital, midazolam, propofol infusion, or high-dose phenobarbital, either alone, or in combination, for < or =146 days. The overall mortality was seven of 22. Mortality was related to etiology, age, and EEG findings. No death occurred in the remote symptomatic group, and three of four younger than 3 years died, whereas only four of 18 older than 3 years died. The mortality rate among patients with focal abnormalities on the EEG was lower than that among those with multifocal or generalized abnormalities. None of the children with normal premorbid neurologic status returned to baseline. CONCLUSIONS: Our data demonstrate a high mortality and morbidity for childhood RSE. Mortality is related to etiology and is higher in younger children and with multifocal or generalized abnormalities on the initial EEG. PMID- 11879351 TI - Progression of postictal to interictal psychosis. AB - PURPOSE: To describe a case series of patients with both postictal psychosis (PIP) and chronic interictal psychosis (IIP). METHODS: We retrospectively reviewed the records of 43 patients with PIP from a comprehensive epilepsy center to find evidence of both PIP and IIP in the same patient. RESULTS: Six (13.9%) of the 43 patients met all the criteria for both PIP and IIP. Five of our six patients had multiple documented PIPs before they became chronically psychotic. The range of length of time between PIP and IIP was 7 to 96 months. Postictal and interictal psychotic behavior was similar or identical in five of six cases. CONCLUSIONS: The results of this study suggest a progression from PIP to IIP: there is a similarity between the symptoms of the two psychoses, a history of multiple PIPs before the first IIP, and a period of months to years between PIP and IIP onset. PMID- 11879352 TI - Peripheral WBC count and serum prolactin level in various seizure types and nonepileptic events. AB - PURPOSE: To analyze effects of different types of seizures and nonepileptic events as well as effects of seizure duration and lapse between the time of seizure and blood collection on serum prolactin level and peripheral white blood cell (WBC) count. METHODS: We prospectively collected blood samples from all patients admitted to our Epilepsy Monitoring Unit at baseline and after an event. Blood samples were analyzed, and serum prolactin level and WBC count were determined. Statistical analyses were performed to evaluate the relation of each type of seizure, its duration, and time lapse between a seizure and collection of blood sample to the serum prolactin level and peripheral WBC count. RESULTS: Serum prolactin level increases above twice the level at baseline after a complex partial seizure or a generalized seizure. Peripheral WBC count is elevated above the upper limit of normal in about one third of cases after a generalized seizure. In generalized seizures, the length of a seizure is positively associated, whereas the lapse time between the seizure onset and blood draw is negatively correlated with the increase in WBC count. Thus the longer the seizure and quicker the blood draw, the higher the WBC count. CONCLUSIONS: We conclude that complex partial or generalized seizures are associated with an increase in serum prolactin level. Peripheral WBC count increases significantly after a generalized seizure and is probably transient in nature. PMID- 11879353 TI - Psychosocial aspects of epilepsy in Oman: attitude of health personnel. AB - PURPOSE: To assess with a questionnaire the awareness and attitudes of the doctors in Oman toward epilepsy. Attitudes of society toward epilepsy have a wide ranging influence, affecting issues as diverse as compliance with treatment and doctor--patient communication. Recent studies in both developing and developed countries suggest that within the medical profession, there is a lack of knowledge and negative attitudes toward people with epilepsy (PWE). There are no equivalent studies for Oman or the Arab world. METHODS: The questionnaire included queries on the backgrounds of the physicians, including their training and qualifications, the main sources of their knowledge of epilepsy, as well as their perceptions of the attributes and care requirements of PWE. RESULTS: Sixty two percent (n=121) of those questioned, who were medical personnel working in different regions of Oman, responded. The results suggest that, despite coming from diverse cultural backgrounds and nationalities, the practicing doctors in Oman gained knowledge of epilepsy much earlier than did their counterparts in developed countries. The majority of the respondents thought that PWE have more propensities toward dysfunctional personality and behavioral characteristics than do "normal" people. On questions relating to public image, our respondents opined that, although the general public is negative toward PWE, the realities regarding PWE should be publicized because PWE are capable of having a normal family life and being an integral part of society. CONCLUSIONS: In spite of having an earlier exposure to seizures and sympathetic acceptance of PWE, negative views still persist on matters related to cognitive and behavior domains. It is concluded that a developing country such as Oman must inculcate more realistic perceptions and attitudes among their doctors toward PWE. PMID- 11879354 TI - Transplacental passage of oxcarbazepine and its metabolites in vivo. AB - PURPOSE: The purpose of this study was to investigate human fetal exposure to oxcarbazepine (OCBZ) in vivo. METHODS: Transplacental passage and placental tissue concentrations of OCBZ and its metabolites were determined. Maternal venous blood, cord blood, and placental tissue samples from 12 mothers using OCBZ during pregnancy alone or in combination with other antiepileptic drugs were collected. Samples were analyzed with high-performance liquid chromatography. RESULTS: Maternal venous concentrations of OCBZ and its major metabolites were at same range as cord blood concentrations (OCBZ in maternal serum, 0.19 +/- 0.16 microg/ml, and in cord serum, 0.21 +/- 0.19 microg/ml; 10-hydroxy-10,11 dihydrocarbamazepine (10-OH-CBZ) in maternal serum, 5.69 +/- 2.49 microg/ml, and in cord serum, 5.23 +/- 1.44 microg/ml; 10,11-trans-dihydroxy-10,11 dihydrocarbamazepine (10,11-D) in maternal serum, 0.29 +/- 0.22 microg/ml, and in cord serum, 0.28 +/- 0.14 microg/ml). OCBZ (0.17 +/- 0.16 microg/g placental tissue), 10-OH-CBZ (3.49 +/- 1.34 microg/g placental tissue) and 10,11-D (0.25 +/ 0.11 microg/g placental tissue) were detected in the placental tissue. The amount of OCBZ detected from placental tissue was 0.01% of the daily dose. CONCLUSIONS: OCBZ, like other antiepileptic drugs, is transferred significantly through the placenta in humans. PMID- 11879357 TI - Reporting clinical trials: full access to all the data. PMID- 11879358 TI - Mechanism of the inhibitory effect of histamine on amygdaloid-kindled seizures in rats. AB - PURPOSE: The mechanism of the inhibitory effect of histamine on amygdaloid kindled seizures was investigated in rats. METHODS: Under pentobarbital anesthesia, rats were fixed to a stereotaxic apparatus, and bipolar electrodes were implanted into the right amygdala. A guide cannula made of stainless steel tubing was implanted into the right lateral ventricle. Electrodes were connected to a miniature receptacle, which was embedded in the skull with dental cement. EEG was recorded with an electroencephalograph; stimulation of the amygdala was applied bipolarly every day by a constant-current stimulator and continued until a generalized convulsion was obtained. RESULTS: Intracerebroventricular (i.c.v.) injection of histamine at doses of 2-10 microg resulted in a dose-related inhibition of amygdaloid-kindled seizures. I.c.v. injection of calcium chloride at doses of 10-50 microg and A23187 at doses of 2-10 microg also caused dose dependent inhibition of amygdaloid-kindled seizures. Calcium chloride at a dose of 10 microg, which showed no significant effect on amygdaloid-kindled seizures when used alone, significantly potentiated the effect of histamine. Similar findings were observed with A23187 at a dose of 2 microg. In addition, EGTA and EGTA/AM antagonized the inhibition of kindled seizures induced by histamine. Moreover, the inhibition of kindled seizures induced by histamine was antagonized by KN62. However, calphostin C did not antagonize the inhibitory effect of histamine. CONCLUSIONS: These results indicated that histamine-induced inhibition of amygdaloid-kindled seizures may be closely associated with a calcium calmodulin-dependent protein kinase II activation pathway. PMID- 11879359 TI - Overexpression of multiple drug resistance genes in endothelial cells from patients with refractory epilepsy. AB - PURPOSE: It has been suggested that altered drug permeability across the blood brain barrier (BBB) may be involved in pharmacoresistance to antiepileptic drugs (AEDs). To test this hypothesis further, we measured multiple drug resistance (MDR) gene expression in endothelial cells (ECs) isolated from temporal lobe blood vessels of patients with refractory epilepsy. ECs from umbilical cord or temporal lobe vessels obtained from aneurysm surgeries were used as comparison tissue. METHODS: cDNA arrays were used to determine MDR expression. MDR protein (MRP1) immunocytochemistry and Western blot analysis were used to confirm cDNA array data. RESULTS: We found overexpression of selected MDR and significantly higher P-glycoprotein levels in "epileptic" versus "control" ECs. Specifically, MDR1, cMRP/MRP2, and MRP5 were upregulated in epileptic tissue, whereas Pgp3/MDR3 levels were comparable to those measured in comparison tissue. The gene encoding cisplatin resistance--associated protein (hCRA-alpha) also was overexpressed in epileptic tissue. Immunocytochemical analysis revealed that MDR1 immunoreactivity was localized primarily in ECs; MRP1 protein levels also were significantly higher in epileptic tissue. CONCLUSIONS: Complex MDR expression changes may play a role in AEDs pharmacoresistance by altering the permeability of AEDs across the BBB. PMID- 11879361 TI - Disparity of perfusion and glucose metabolism of epileptogenic zones in temporal lobe epilepsy demonstrated by SPM/SPAM analysis on 15O water PET, [18F]FDG-PET, and [99mTc]-HMPAO SPECT. AB - PURPOSE: To elucidate uncoupling of perfusion and metabolism and its significance in epilepsy, 15O water and 18F fluorodeoxyglucose (FDG) positron emission tomography (PET) and Tc-99m hexamethyl-propyleneamine-oxime (HMPAO) single-photon emission computed tomography (SPECT) were examined by SPM (statistical parametric mapping) and quantitation by using SPAM (statistical probabilistic anatomic map). METHODS: [15O]water and [18F]FDG-PET, and [99mTc]-HMPAO SPECT were performed in 25 patients (SPECT in 17 of 25) with medial temporal lobe epilepsy. For volume of interest (VOI) count analysis, the normalized counts using VOI based on SPAM templates of PET and SPECT were compared with those of the normal controls. Perfusion or metabolism was found abnormal if the Z score was >2 for each VOI. For SPM analysis, the differences between each patient's image and a group of normal control images (t statistic for p < 0.01) on a voxel-by-voxel basis were examined to find significant decreases in perfusion or metabolism. RESULTS: With SPAM VOI count analysis, areas of hypoperfusion were found in 13 patients in the epileptogenic temporal lobes by [15O]water PET and areas of hypometabolism in 21 patients by [18F]FDG-PET. With voxel-based SPM analysis, the epileptogenic zones were localized in 15 by [15O]water PET and in 23 patients by [18F]FDG-PET. The localization by [15O]water PET was concordant with that of [18F]FDG-PET. The areas of hypoperfusion on [15O]water PET were absent or smaller than the areas of hypometabolism on [18F]FDG-PET. Interictal [99mTc]-HMPAO SPECT revealed the hypoperfused zones in seven of 17 patients on visual assessment. CONCLUSIONS: SPAM VOI count and SPM analysis of [15O]water and [18F]FDG-PET and [99mTc]-HMPAO SPECT revealed that in the same patients, the areas of hypoperfusion were concordant with but smaller than the areas of hypometabolism. Discordance of perfusion and metabolic abnormalities represents an uncoupling of perfusion and metabolism in the epileptogenic zones, and this might explain the lower diagnostic accuracy of perfusion imaging in temporal lobe epilepsy. PMID- 11879362 TI - MEG predicts epileptic zone in lesional extrahippocampal epilepsy: 12 pediatric surgery cases. AB - PURPOSE: To discover whether the spatial distribution of spike sources determined by magnetoencephalography (MEG) provides reliable information for planning surgery and predicting outcomes in pediatric patients with lesional extrahippocampal epilepsy. METHODS: We retrospectively studied 12 children with extrahippocampal epilepsy secondary to cortical dysplasia (CD), tumor, or porencephalic cyst. We compared interictal MEG spike source locations and somatosensory evoked fields derived from equivalent-current dipole modeling with intraoperative or extraoperative electrocorticography (ECoG). RESULTS: MEG spike sources were found in proximity to the lesion in all patients and extended from lesions in five patients with CD. Marginal spike sources were noted in three patients with tumors, one patient with a cyst, and one with CD, and extramarginal sources in three patients with tumors. Three patients with tumors underwent lesionectomy only; two had further cortical excisions. One patient with CD underwent lesionectomy only, three had lesionectomy and cortical excisions, and two had lesionectomy and multiple subpial transection. Asymmetric MEG spike sources correlated with ECoG findings in all patients. Residual epileptiform discharges on postexcisional ECoG corresponded to spike sources in three patients with tumors and one patient with a cyst. Eleven patients have been seizure free for 1-6 years (mean, 4 years). One patient had residual seizures after incomplete excision of right temporal CD. CONCLUSIONS: MEG delineated asymmetric epileptogenicity surrounding lesions and the eloquent cortex. Complete tumor resection produced favorable outcomes despite residual postexcisional ECoG spikes and extramarginal MEG spike sources. CD characterized by clusters of MEG spike sources within and extending from lesions seen on magnetic resonance imaging (MRI) should be removed to prevent seizures. PMID- 11879363 TI - Diagnostic performance of [18F]FDG-PET and ictal [99mTc]-HMPAO SPECT in occipital lobe epilepsy. AB - PURPOSE: We investigated whether interictal F-18 fluorodeoxyglucose positron emission tomography ([18F]FDG-PET) or ictal [99mTc]-HMPAO single-photon emission computed tomography (SPECT) was useful to find epileptogenic zones in occipital lobe epilepsy (OLE). METHODS: We reviewed visually and quantified patterns of hypometabolism in interictal [18F]FDG-PET and those of hyperperfusion in ictal SPECT in 17 OLE patients (27 plus minus 6.8 years old; M/F, 10/7; injection time, 30 plus minus 17 s). OLE was diagnosed based on invasive electroencephalography, surgery, and postsurgical outcome (Engel class I in all at an average of 26 months after surgery). RESULTS: Epileptogenic zones were correctly localized in nine (60%) of 15 patients by interictal [18F]FDG-PET, and asymmetric indices corroborated visual diagnosis. Epileptogenic hemispheres were correctly lateralized in 14 (93%) of 15 patients on [18F]FDG-PET. Epileptogenic hemispheres were correctly lateralized in 13 (76%) of 17 patients using ictal SPECT, but localization was possible in only five (29%) patients. Interictal [18F]FDG-PET was helpful in two of the patients who showed no abnormality on magnetic resonance imaging (MRI) and no possible localization with ictal SPECT. CONCLUSIONS: In OLE, ictal SPECT was helpful in lateralization, but less helpful in localization. Interictal [18F]FDG-PET was helpful in localization or lateralization of epileptogenic zones, even in patients with ambiguous MRI or ictal SPECT findings. PMID- 11879360 TI - Proton spectroscopic imaging shows abnormalities in glial and neuronal cell pools in frontal lobe epilepsy. AB - PURPOSE: Proton magnetic resonance spectroscopic imaging (1H MRSI) can lateralize the epileptogenic frontal lobe by detecting metabolic ratio abnormalities in frontal lobe epilepsy (FLE). We used 1H MRS to lateralize and localize the epileptogenic focus, and we also sought to characterize further the metabolic abnormality in FLE. METHODS: We measured signals from N-acetyl aspartate (NAA), choline-containing compounds (Cho), and creatine + phosphocreatine (Cr) in the supraventricular brain of 14 patients with frontal or frontoparietal epilepsy and their matched controls. The supratentorial brain also was segmented into gray matter, white matter, and cerebrospinal fluid classes. Regional metabolite alterations were compared with localizing and lateralizing results from other examination modalities and with histology from three patients. RESULTS: Spectroscopy lateralized the epileptogenic focus in 10 patients in agreement with video-EEG and functional imaging. In four patients, spectroscopy showed bilateral, focal metabolic abnormality, whereas video-EEG suggested unilateral or midline abnormality. In the epileptogenic focus, Cho and Cr were increased by 23% and 14%, respectively, and NAA was decreased by 11%, suggesting metabolic disturbances both in the glial and in the neuronal cell pools. Two Taylor dysplasia lesions confirmed by histology and one with radiologic diagnosis showed high Cho and low or normal NAA, whereas two dysembryoplastic neurogenic tumors had normal Cho and low NAA. Contralateral hemisphere NAA/(Cho + Cr) was decreased in FLE, indicating diffusely altered brain metabolism. Segmentation of brain tissue did not reveal atrophic changes in FLE. CONCLUSIONS: Spectroscopy is useful in lateralizing frontoparietal epilepsy and shows promise as a "noninvasive biopsy" in epileptogenic lesions. PMID- 11879364 TI - Familial infantile myoclonic epilepsy: clinical features in a large kindred with autosomal recessive inheritance. AB - PURPOSE: To describe the clinical features of a large kindred with familial infantile myoclonic epilepsy (FIME) with autosomal recessive inheritance, and to discuss the nosology of the early infantile myoclonic epilepsies (IMEs). METHODS: The family descends from the intermarriage of two couples of siblings. In a previous study, we mapped the genetic locus to chromosome 16p13. We analyzed results of family records and personal history, psychomotor development, neurologic examination, epilepsy features, and EEG recordings for each subject. RESULTS: FIME has a strong penetrance (eight affected of 14 subjects) and a homogeneous clinical picture. Like the benign form of infantile myoclonic epilepsy (BIME), FIME is a true idiopathic IME with unremarkable history, no neurologic or mental impairment, good response to treatment, and normal interictal EEG pattern. Conversely, onset with generalized epileptic seizures without fever (four patients) or with fever (one patient), frequency and duration of the myoclonic seizures, occurrence of generalized tonic--clonic seizures (GTCSs) in all patients and persistence of seizures into adulthood are characteristics of the severe infantile myoclonic epilepsy (SIME). CONCLUSIONS: Clinical overlap probably exists among the myoclonic epilepsies of infancy. FIME differs from other forms of IME in its phenotypic features. The peculiar mode of inheritance is explained by the genetic background of the family. Genetic studies suggest linkage to chromosome 16 in familial cases of true IME. PMID- 11879365 TI - Benign partial epilepsies of adolescence: a report of 37 new cases. AB - PURPOSE: To delineate the electroclinical features of patients with partial seizures in adolescence with a benign outcome. METHODS: Patients were recruited in five different Italian epilepsy centers. Patients were selected among those with partial seizures between ages 11 and 17 years. We excluded benign childhood epilepsies, those with neurologic or mental deficits, and those with neuroradiologically documented lesions. We also excluded patients with less than 3 years' follow-up or who were still receiving antiepileptic therapy. RESULTS: There were 37 (22 male, 15 female) patients. Seizures started at the mean age of 14.5 years (range, 11-16.11). Two main electroclinical patterns emerged: 16 of 37 patients had somatomotor seizures frequently associated with focal theta discharges involving the centroparietal regions. Ten of 37 patients showed versive seizures and interictal spiking involving the posterior regions. A third group had clinical characteristics resembling the cases described by Loiseau. All had a favorable outcome. CONCLUSIONS: This relevant multicenter study further confirms the existence of benign partial epilepsies with onset during adolescence. PMID- 11879366 TI - Two-year remission and subsequent relapse in children with newly diagnosed epilepsy. AB - PURPOSE: Although remission is the ultimate measure of seizure control in epilepsy, and epilepsy syndrome should largely determine this outcome, little is known about the relative importance of syndrome versus other factors traditionally examined as predictors of remission or of relapse after remission. The purpose of this study was to examine remission and relapse with respect to the epilepsy syndrome and other factors traditionally considered with respect to seizure outcome. METHODS: A prospectively identified cohort of 613 children with newly diagnosed epilepsy was assembled and is actively being followed to determine seizure outcomes. Epilepsy syndrome and etiology were classified at diagnosis and again 2 years later. Remission was defined as 2 years completely seizure-free, and relapse as the recurrence of seizures after remission. Multivariable analysis was performed with the Cox proportional hazards model. RESULTS: Five hundred ninety-four of the original 613 children were followed > or = 2 years (median follow-up, 5 years). Remission occurred in 442 (74%), of whom 107 (24%) relapsed. On multivariable analysis, idiopathic generalized syndromes and age at onset between 5 and 9 years were associated with a substantially increased remission rate, whereas remote symptomatic etiology, family history of epilepsy, seizure frequency, and slowing on the initial EEG were associated with a decreased likelihood of attaining remission. Young onset age (<1 year) and seizure type were not important after adjustment for these predictors. Relapses occurred more often in association with focal slowing on the initial EEG and with juvenile myoclonic epilepsy. Benign rolandic epilepsy and age at onset <1 year were associated with markedly lower risks of relapse. About one fourth of relapses were apparently spontaneous while the child was taking medication with good compliance, and more than half occurred in children who were tapering or had fully stopped medication. CONCLUSIONS: A large proportion of children with epilepsy remit. Symptomatic etiology, family history, EEG slowing, and initial seizure frequency negatively influence, and age 5-9 years and idiopathic generalized epilepsy positively influence the probability of entering remission. Factors that most influence relapse tend to be different from those that influence remission. PMID- 11879367 TI - Seizure semiology and neuroimaging findings in patients with midline spikes. AB - PURPOSE: Midline epileptiform discharges are rare compared with discharges at other scalp locations. Neuroimaging results and semiologic seizure characteristics of patients with midline spikes are not adequately described. The aim of this study was to describe the neuroimaging findings and detailed seizure semiologies in patients with midline spikes. METHODS: We reviewed the EEG database of the University of Michigan Medical Center and identified 35 patients with midline spikes. Information about seizure types and neuroimaging results was obtained from a review of medical records. The seizures were classified according to the International League Against Epilepsy (ILAE) criteria and semiologic classification. RESULTS: Twenty-nine (83%) patients had a history of seizures. Complex partial seizures and simple partial seizures were the most common seizure types, experienced by 66% of patients. The age at seizure onset was within the first 10 years in 90% of patients. According to the semiologic seizure classification, automotor seizures and tonic seizures were the most common seizure types. Neuroimaging studies were abnormal in 45% of patients. When focal abnormalities were detected, they were lateralized to one of the frontal lobes in all cases. CONCLUSIONS: Our results indicate that in the majority of patients, midline spikes represent focal epileptiform activity rather than fragments of generalized discharges, and are most commonly associated with seizures of partial onset. Automotor seizures and tonic seizures are the most common semiologies. Focal radiologic abnormalities tend to be lateralized to one of the frontal lobes. PMID- 11879368 TI - The effects of lamotrigine on sleep in patients with epilepsy. AB - PURPOSE: The older antiepileptic drugs (AEDs) have a variety of effects on sleep, including marked reduction in REM, slow-wave sleep (SWS) and sleep latency, and increased percentage of light sleep. The effects of the newer AEDs on sleep are unknown. Our purpose was to study the effect of lamotrigine (LTG) on sleep. METHODS: Ten adults with focal epilepsy, in whom the decision was made to add LTG to either phenytoin (PHT) or carbamazepine (CBZ) for control of seizures, were the subjects of this study. Patients underwent pre- and posttreatment polysomnography (PSG) and completed sleep questionnaires. Polygraphic variables and Epworth Sleepiness Scale (ESS) scores, a subjective measure of sleep propensity, were compared by using the Wilcoxon sign rank test. RESULTS: Seven patients were taking CBZ, and three were treated with PHT. All subjects were titrated to an LTG dose of 400 mg/day. Treatment with LTG produced a significant decrease in SWS and an increase in stage 2 sleep percentage. No significant difference in ESS or any of the other polygraphic variables was observed. However, LTG treatment was associated with a reduction in arousals and stage shifts and an increase in REM periods. No subjects reported insomnia with treatment. CONCLUSIONS: LTG appears to be less disruptive to sleep than some of the older AEDs. PMID- 11879369 TI - Pharmacokinetic study of levetiracetam in children. AB - PURPOSE: The pharmacokinetics of the novel antiepileptic drug (AED) levetiracetam and its major metabolite, ucb L057, were studied in children with partial seizures in a multicenter, open-label, single-dose study. METHODS: Twenty-four children (15 boys, nine girls), 6 to 12 years old, received a single dose of levetiracetam (20 mg/kg) as an adjunct to their stable regimen of a single concomitant AED, followed by a 24-h pharmacokinetic evaluation. RESULTS: In children, the half-lives of levetiracetam and its metabolite ucb L057 were 6.0 +/ 1.1 and 8.1 +/-2.7 hours, respectively. The Cmax and area under the curve (AUC) of levetiracetam equated for a 1-mg/kg dose were lower in children (Cmax, norm=1.33 plus minus 0.35 microg/ml; AUCnorm=12.4 +/- 3.5 microg/h/ml) than in adults (Cmax, norm=1.38 +/- 0.05 microg/ml; AUCnorm=11.48 +/- 0.63 microg/h/ml), whereas the renal clearance was higher. The apparent body clearance (1.43 +/- 0.36 ml/min/kg) was approximately 30-40% higher in children than in adults. Levetiracetam was generally well tolerated. CONCLUSIONS: On the basis of these data, a daily maintenance dose equivalent to 130-140% of the usual daily adult maintenance dosage (1,000-3,000 mg/day) in two divided doses, on a weight normalized level (mg/kg/day) is initially recommended. Clinical efficacy trials in children are ongoing with dosages of 20 to 60 mg/kg/day. PMID- 11879370 TI - Infantile spasms in Down syndrome: good response to a short course of vigabatrin. AB - PURPOSE: To evaluate the efficacy of vigabatrin (VGB) in the treatment of infantile spasms (ISs) associated with Down syndrome (DS) and to assess the feasibility of early discontinuation to reduce the possible retinal toxicity. METHODS: Five children with ISs with DS were treated with vigabatrin as first line monotherapy in an open prospective study. The short-term response was evaluated, and VGB was continued in responders. The treatment was stopped after 6 months in children who were still spasm free. RESULTS: Four children of five became spasm free with VGB, three of them responding within 1 week. This response was maintained during the 6 months of VGB treatment. After VGB discontinuation, and with a follow-up ranging from 2 to 4 years, none of the responders experienced spasm recurrence or other types of seizures. CONCLUSIONS: This study confirms the efficacy of VGB in ISs associated with DS. Moreover, it shows that the duration of VGB treatment can be reduced to 6 months without relapse of ISs. This short treatment might reduce the risk of developing visual field constriction. PMID- 11879371 TI - Premature ovarian failure in women with epilepsy. AB - PURPOSE: Women with epilepsy (WWE) have an increased risk for several reproductive endocrine disorders that may affect their fertility. The incidence of premature ovarian failure (POF) in women with epilepsy has not been systematically studied. This study examined the incidence of POF in women with epilepsy. METHODS: Fifty consecutively evaluated cognitively normal women with epilepsy, aged 38-64 years, whose seizures began before age 41 years, were interviewed for symptoms of perimenopause and menopause. Endocrine studies, performed in women aged 45 years or younger at the time of evaluation, included serum follicle-stimulating hormone (FSH; done on menstrual cycle day 3 in menstruating women), inhibin A levels when FSH was normal, thyroid-stimulating hormone (TSH), prolactin, and, in menstruating women, menstrual cycle day 20 serum progesterone level. Nonsurgical premature menopause was defined as secondary amenorrhea of >12 months' duration with FSH levels of >14 International Units (IU) in women younger than 42 years. Premature perimenopause was defined by the presence of one or more of the following: somatic perimenopausal symptoms; change in previously regular menstrual cycles without evidence of other reproductive endocrine disturbance; and FSH level of >14 IU or inhibin A level of <7 pg/ml. Similarly aged neurologically normal women seen in the menopause and sleep clinics served as control subjects. Statistical analysis included Fisher's exact test, Kruskal-Wallis test, t test, and multivariate logistic regression analysis with significance set at p < 0.05. RESULTS: Seven (14%) of 50 women with epilepsy had nonsurgical premature perimenopause (six of seven) or menopause (one of seven), compared with three of 82 control (p=0.042). Five of 41 women with localization-related epilepsy (LRE) had POF compared with two of nine women with primary generalized epilepsy (PGE; p=0.595). Mean age of POF was 39.6 years (range, 37-42 years). Seizure duration, age at seizure onset, seizure severity and lateralization, smoking history, age of menarche, body mass index and incidence of depression was not statistically different between women with and without POF. There was no statistically significant association between POF and antiepileptic drugs (AEDs). Women with POF were more likely to have had catamenial exacerbation of their seizures than were women without POF (p=0.02). CONCLUSIONS: Women with epilepsy have an increased risk for developing POF. This finding should be considered in counseling women with epilepsy on family planning. PMID- 11879372 TI - Seizures in the critically ill: the role of imipenem. AB - PURPOSE: To determine the risk of seizures in critically ill patients receiving the antibiotic imipenem, a broad-spectrum antibiotic that has been associated with seizures. Reports generally have not considered other contributing factors such as dose, seizure history, and morbidity index of the underlying illness necessitating the antibiotic. METHODS: Charts of all patients in a 450-bed municipal hospital who received imipenem in a 6-month period, as determined by pharmacy records, were reviewed for dosage and duration of imipenem use, occurrence of seizures. and mortality outcome. Attention was paid to demographic features; pattern of seizure occurrence during, before, and after imipenem use; renal function; and correction for dosage based on size. RESULTS: Seventy-five charts were reviewed. Sixty-three patients had no seizures during the hospitalization, four had seizures while receiving imipenem, and eight had seizures during the hospitalization but before or after imipenem use. The incidence of seizures was 4/1,000 patient-days on, and 3.9/1,000 patient-days off imipenem (not significant). The risk of seizure in both groups was considerably higher in those patients with a history of seizures before hospitalization. The presence of other factors that could contribute to increased concentration of imipenem in the brain, such as renal failure or acute stroke, did not contribute to seizure incidence. Metabolic derangement, anoxia, and phenytoin discontinuation did contribute to seizure incidence. CONCLUSIONS: Seizure incidence is increased in all critically ill patients (16% of patients studied), but with no added risk during the period patients received imipenem. Determining the proper dose based on a patient's body mass, correction of dose in the presence of renal failure, and avoidance of excess of 2 g/day of imipenem removes any added risk for seizures from imipenem. Despite experimental data to suggest action of imipenem on the glutamate/N-methyl-d-aspartate receptor, or interference with binding to the gamma-aminobutyric acid receptor, and early clinical studies that warned against its use because of seizure risk, we found that careful use of this antibiotic is safe. PMID- 11879373 TI - Prospective study of seizures in the elderly in the Marshfield Epidemiologic Study Area (MESA). AB - PURPOSE: To assess the incidence, etiology, antiepileptic drug (AED) use, and quality of life in patients having their first seizure at age 50 years or older. METHODS: All patients in the Marshfield Epidemiologic Study Area (MESA) aged 50 years or older having their first seizure between July 1, 1996, and June 30, 1998, were identified through the diagnostic coding system. Patients were followed up for 12 months for recurrent seizures. Etiologies, drug treatments, magnetic resonance imaging (MRI), computed tomography (CT), or EEG results and medication adverse effects were recorded. Those patients who reached 1-year follow-up received a quality of life (QOLIE-31) survey. RESULTS: Forty-eight patients having a first seizure were identified (162 of 100,000). Twelve patients had had recurrent seizures, and 36 had a single seizure at the time of study entry. Fourteen of these 36 had had an abnormal MRI, CT, or EEG. The remaining 22 had a single seizure and normal imaging and EEG. Six of these had one or more subsequent seizures, and all six were in the group with normal tests. Etiologies included vascular, neoplasm, trauma, dementia, metabolic, and unknown. Seventy five percent of the patients achieved seizure control with phenytoin, carbamazepine, and/or valproate. Twenty-seven percent experienced adverse side effects. None had been given second-generation AEDs as an initial treatment. Thirty-one patients received the QOLIE-31 survey; 20 did not complete the survey for various reasons. The mean QOLIE-31 scores for those completing the survey were significantly higher than those of the reference cohort. CONCLUSIONS: The incidence of first seizure in MESA is high in the elderly and increases with advancing age. As in other studies, vascular causes accounted for the largest etiology. Use of new AEDs was uncommon. There was a high incidence of untoward side effects related to the traditional AEDs. High morbidity and mortality unrelated to seizures limits follow-up analysis in the elderly. Quality-of-life analysis via QOLIE questionnaires is problematic in this population. PMID- 11879374 TI - Impaired motor function in patients with psychogenic pseudoseizures. AB - PURPOSE: To evaluate motor speed and grip strength in patients with well documented psychogenic pseudoseizures. METHODS: We analyzed manual motor speed and grip strength in a group of 40 patients with confirmed psychogenic pseudoseizures (without evidence of concomitant epilepsy) and a group of 40 normal controls matched for handedness and gender, and of comparable age. The two groups were compared with respect to manual motor performance with the dominant hand, nondominant hand, and asymmetry between the dominant and nondominant hands. For the patient sample, we reviewed the neurologic history. RESULTS: Patients with pseudoseizures performed more poorly than controls with both dominant and nondominant hands. In addition, pseudoseizure patients failed to demonstrate the dominant-hand advantage observed in the normal control subjects on both tasks. The patient group had a high incidence of head trauma and other antecedent neurologic risk factors, and the proportion of left-handers was 3 times higher than expected. CONCLUSIONS: Bilaterally reduced motor speed and grip strength, reduced intermanual performance asymmetry, the high percentage of left-handers, and historical evidence of antecedent insults to the brain indicate that frontal lobe impairment may be common in patients with psychogenic pseudoseizures. PMID- 11879375 TI - Ring 20 chromosome syndrome with epilepsy and dysmorphic features: a case report. AB - Relatively few cases of the 20 ring chromosome [r(20)] syndrome have been reported. Epileptic seizures, behavioral problems, mental retardation, and absence of definite dysmorphic features characterize this syndrome. We present a patient with the classic genetic and phenotypic findings. A 42-month-old boy with mild dysmorphic features and psychomotor retardation has had generalized tonic clonic seizures, resistant to antiepileptic drug therapy since he was 26 months old. Electroencephalography (EEG) was performed on several occasions, as were brainstem auditory evoked potentials (BAEPs), magnetic resonance imaging (MRI), and cytogenetic studies. The EEG showed slow waves in anterior regions intermingled with spikes in temporal areas. The BAEPs were abnormal, and neuroimaging studies were normal. The chromosome r(20) appeared in 100 metaphases studied. Parental chromosomes were of normal karyotype. The genetic and EEG finding from this patient strongly suggest that epilepsy associated with 20 ring chromosome syndrome is a distinct new entity, although the clinical manifestations may be broader than previously recognized. PMID- 11879376 TI - Levetiracetam psychosis in children with epilepsy. AB - PURPOSE: Levetiracetam is a new anticonvulsant (AED) with a novel mechanism of action. Although it is generally well tolerated with a good cognitive profile, irritability and hostility have been reported in some adults taking levetiracetam. Observations in children are limited; levetiracetam is not yet approved by the Food and Drug Administration for use in children. METHODS: In four young patients, acute psychosis developed within days to months of initiation of levetiracetam for seizures. RESULTS: A 5-year-old girl began having visual hallucinations of spiders in her room 14 days after starting levetiracetam. A 13-year-old boy began having auditory hallucinations, insomnia, and screaming behavior 3 months after initiation of levetiracetam. A 16-year-old girl became acutely agitated, hyperreligious, and had persecutory delusions within 7 days of starting levetiracetam. A 17-year-old girl had auditory hallucinations telling her to sing and yell after 30 days of taking the drug. All four children had dramatic improvement within days of either discontinuing or decreasing the dose of levetiracetam. The three adolescents had historical findings consistent with mild behavioral problems before initiating levetiracetam, and all four patients had prior cognitive deficits. CONCLUSIONS: Reversible treatment-emergent psychosis associated with levetiracetam therapy was observed in four children and adolescents. Whether rapid initiation or prior neurobehavioral problems predispose to this side effect is not established. PMID- 11879377 TI - Placebos, placebo effect, and the response to the healing situation: the evolution of a concept. AB - In spite of its impressive progress, medicine has been strongly criticized for relying on its modern biomedical tradition to the neglect of the psychosocial aspects of health. This neglect may account for patients' dissatisfaction and eventual use of alternative health approaches. The concept of placebo has sustained dramatic "protean" metamorphoses through the ages. For centuries, placebos have been regarded as powerful deceptive therapies. From the middle of the twentieth century, however, conventional medicine has used placebos as methodologic tools to distinguish the specific from the nonspecific ingredients in treatments. In modern medical research, the double-blind, placebo-controlled, randomized clinical trial has been established as the gold standard for the assessment of any new treatment. Recently a new trend regarding placebos seems to have emerged. The placebo and other nonspecific effects elicited by the "healing situation" have been independently subjected to scientific study. Progress in this area may promote useful clinical applications, enabling physicians to broaden their perspectives on the healing process. We present the historical changes of the concept of placebo and the ethical issues raised by their use. PMID- 11879380 TI - Brain sterols in the AY-9944 rat model of atypical absence seizures. AB - PURPOSE: The AY-9944 (AY)-treated rat is a reproducible and clinically relevant animal model of atypical absence seizures. AY inhibits cholesterol synthesis, but the relation between brain sterol levels and the spontaneously recurrent absence seizures has not been determined. METHODS: Long-Evans hooded rats were treated every 6 days from postnatal day (P)2 to P20 with AY (7.5 mg/kg, s.c.) or saline. Electrodes were permanently implanted under pentobarbital anesthesia at P50. Spike-and-wave discharge (SWD) duration and amplitude were quantified at P55. Changes in brain sterols after AY were examined in three different experiments, looking at brain regions (experiment 1), recovery after stopping AY (experiment 2), or gender differences (experiment 3). RESULTS: Experiment 1: AY caused spontaneously recurrent slow SWD that lasted 59 times longer and had a 3.2-fold higher amplitude than that in controls. At P55, brain cholesterol was reduced and 7-dehydrocholesterol was increased in all brain regions (p < 0.0001). Experiment 2: Four hundred days after stopping AY-9944 treatment (P420), brain sterol levels had returned to normal levels, but the AY-induced SWD lasted twice as long as at P55. Experiment 3: At P55, AY-induced changes in plasma and liver (but not brain) sterols were significantly more severe in females compared with males. CONCLUSIONS: AY-induced seizures appear to be related to AY-induced changes in brain sterols but persisted long after the sterols had returned to normal after the last AY injection. Hence, there appears to be a critical developmental window during which the AY must be given but after which the AY-induced change in brain sterols is no longer essential to sustaining the seizures. PMID- 11879381 TI - Selective blockade of N-type calcium channels by levetiracetam. AB - PURPOSE: We investigated the effect of the new antiepileptic drug (AED) levetiracetam (LEV) on different types of high-voltage-activated (HVA) Ca2+ channels in freshly isolated CA1 hippocampal neurons of rats. METHODS: Patch clamp recordings of HVA Ca2+ channel activity were obtained from isolated hippocampal CA1 neurons. LEV was applied by gravity flow from a pipette placed near the cell, and solution changes were made by electromicrovalves. Ca2+ channel blockers were used for separation of the channel subtypes. RESULTS: The currents were measured in controls and after application of 1-200 microM LEV. LEV irreversibly inhibited the HVA calcium current by approximately 18% on the average. With a prepulse stimulation protocol, which can eliminate direct inhibition of Ca2+ channels by G proteins, we found that G proteins were not involved in the pathways underlying the LEV inhibitory effect. This suggested that the inhibitory effect arises from a direct action of LEV on the channel molecule. The blocking mechanism of LEV was not related to changes in steady state activation or inactivation of Ca2+ channels. LEV also did not influence the rundown of the HVA Ca2+ current during experimental protocols lasting approximately 10 min. Finally, LEV at the highest concentration used (200 microM) did not influence the activity of L-, P- or Q-type Ca2+ channels in CA1 neurons, while selectively influencing the activity of N-type calcium channels. The maximal effect on these channels separated from other channel types was approximately 37%. CONCLUSIONS: Our results provide evidence that LEV selectively inhibits N-type Ca2+ channels of CA1 pyramidal hippocampal neurons. These data suggest the existence of a subtype of N-type channels sensitive to LEV, which might be involved in the molecular basis of its antiepileptic action. PMID- 11879382 TI - Randomized double-blind parallel-group study comparing cognitive effects of a low dose lamotrigine with valproate and placebo in healthy volunteers. AB - PURPOSE: This study aimed at investigating the cognitive and mood effects of lamotrigine (LTG) versus valproate (VPA) and placebo (PBO). METHODS: By studying the effects in healthy volunteers, it is possible to separate the genuine effects of LTG from the cognitive improvements, caused by better seizure control. The study used a pretest-posttest comparison of 50 mg LTG, 900 mg VPA, or PBO in a double-blind single-dummy parallel-group design with 30 healthy volunteers. Study duration was 12 days (with a last control on day 13). Outcome measures included cognitive tests (FePsy neuropsychological test battery), mood scales (ASL; mood rating scale), and a scale for subjective complaints (ABNAS Neurotoxicity scale). Total sleep time was controlled with actigraphic recordings. The results were analyzed by comparing the change over time (pretest with posttest) for the three treatments with Student's t tests. RESULTS: COGNITIVE TESTS: significant differences between the treatments were found for measurements of cognitive activation (i.e., three of the four simple reaction-time measurements showed statistically significant differences in change between PBO and LTG in favor of LTG (p=0.03; 0.03; 0.04); two of four tests showed statistically significant differences in change between LTG and VPA, both in favor of LTG (p=0.03; 0.05). SUBJECTIVE COMPLAINTS: the ABNAS-neurotoxicity scale reveals a significant reduction of drug-related cognitive complaints for the subjects taking LTG, relative to VPA (p=0.02). MOOD RATING: significant changes were found on the scale assessing "tiredness," showing increased tiredness/sedation for VPA relative to PBO (p=0.02) and on the "timid scale" for LTG reporting "being more at ease" compared with both PBO and VPA (p=0.02; 0.02). The general direction of change for the mood scales was toward "activation" for LTG (five of six scales improved), whereas for VPA, the reverse effect was found (four of six scales showed a change in the direction of "tiredness/sedation"). CONCLUSIONS: Short term treatment in normal volunteers with a low dose of LTG resulted in improved cognitive activation on simple reaction-time measurements, a more positive subjective report about the impact of drug treatment relative to VPA, and mood changes concurring with the activating effect demonstrated by the cognitive tests. PMID- 11879383 TI - Focal cortical dysplasias in eloquent cortex: functional characteristics and correlation with MRI and histopathologic changes. AB - PURPOSE: Focal cortical dysplasia (CD) is increasingly recognized as a common pathologic substrate of medically intractable epilepsy. As these lesions are often localized in the frontal lobe (therefore in potentially eloquent cortex), an understanding of the functional status of the involved region(s) and of its anatomic and pathologic correlates is of prime importance. The purpose of this study is to assess the function of focal CD in relation to magnetic resonance imaging (MRI) and histopathologic features. METHODS: Eight patients operated on for medically intractable epilepsy with histologically proven focal CD involving putative eloquent cortex in the frontal lobe (perirolandic and Broca's areas) were included in the study. Functional regions (motor and language) and epileptogenic areas were assessed by extraoperative electrocorticographic recording and electrical cortical mapping. Cortical functions were correlated with the extent of epileptogenicity on electrocorticographic recordings, MRI features, and histologic characteristics. RESULTS: Language or motor areas were colocalized with epileptogenic regions (n=6 of 8, 75%), but were not mapped in regions of increased signal on fluid-attenuated inversion recovery (FLAIR) MRI (when they were identified) on preoperative MRI (n=5 of 5, 100%). Histologically, balloon cells were almost exclusively found in nonfunctional regions with FLAIR MRI abnormalities. When resected, regions of motor cortex were characterized by cortical dyslamination, columnar disorganization, and dysmorphic neurons, but were devoid of balloon cells. CONCLUSIONS: We found an absence of language or motor functions in perirolandic and Broca's areas that showed decreased epileptogenicity, histopathological evidence of CD with balloon cells and FLAIR MRI signal increase. Language and motor functions were present in epileptogenic and dysplastic areas with no balloon cells and no FLAIR signal abnormalities. These findings have implications on options for epilepsy surgery in patients with CD. PMID- 11879384 TI - Focal cortical dysplasia of Taylor's balloon cell type: a clinicopathological entity with characteristic neuroimaging and histopathological features, and favorable postsurgical outcome. AB - BACKGROUND AND PURPOSE: Focal cortical dysplasia of Taylor's balloon-cell type (FCD-BC) are a frequent cause of pharmacoresistant epilepsy in young patients. In order to characterize FCD-BC, we coupled MRI and histopathology, and analyzed the clinical outcome following epilepsy surgery. METHODS: From an epilepsy data bank with 547 histological specimens, 17 FCD-BC were re-evaluated of which high resolution MRI was available. Five additional FCD-BC were prospectively identified by MRI. Histopathological and immunohistochemical features were related to MRI. Outcome following lesionectomy was analyzed as determined on routine examinations 3, 6 and 12 months following surgery. RESULTS: All but one lesion were located outside the temporal lobe. A markedly hyperintense funnel shaped subcortical zone tapering towards the lateral ventricle was the characteristic finding on FLAIR MRI. Histopathologically, the subcortical zone of the FCD-BC displayed hypomyelinated white matter with radially oriented balloon cells and gliosis. Dysplastic neurons were found in the adjacent, disorganized cortex. All patients with complete lesionectomy were seizure free one year following surgery. CONCLUSION: Focal cortical dysplasias of Taylor's balloon-cell type (FCD-BC) have characteristic MRI and histopathological findings. MRI recognition is important, since outcome following resective surgery is favorable. PMID- 11879385 TI - Autonomic auras: left hemispheric predominance of epileptic generators of cold shivers and goose bumps? AB - PURPOSE: Autonomic seizures in temporal lobe epilepsies associated with "cold shivers and goose bumps" as a principal ictal sign or aura have only rarely been studied. METHODS: Sixteen patients with autonomic auras [cold shivers or cold sweats (n=11), goose bumps (n=4), one patient showed both ictal signs] were analyzed. RESULTS: Lesions were detected in 12 patients. The etiology was heterogeneous [cryptogenic, arteriovenous (AV)-malformation cyst, trauma, hippocampal sclerosis]. Eight patients underwent epilepsy surgery. The localization and lateralization of the focal epileptic activity in the temporal lobes was determined either by magnetic resonance imaging lesions, EEG, ictal signs, or single-photon emission computed tomography imaging. CONCLUSIONS: The patients with temporal lobe epilepsies associated with "cold shivers and/or goose bumps" showed a left hemispheric predominance of the focal abnormality in the temporal lobe. PMID- 11879386 TI - Interictal epileptiform discharges do not change before seizures during sleep. AB - PURPOSE: Whether interictal epileptiform discharges (IEDs) increase, decrease, or are unchanged before epileptic seizures has implications for the pathophysiology of epilepsy. Prior studies relating IEDs and seizures have not demonstrated a change in IEDs before seizures. However, they have not controlled for changes in the depth of sleep. Our objective was to test the hypothesis that IEDs are related to seizures during sleep while adjusting for log delta power (LDP), a continuous measure of sleep depth. METHODS: Twenty-two seizures during sleep were identified in 16 subjects with epilepsy admitted for presurgical monitoring. The IEDs that occurred in the hour of sleep before each seizure were used to test the relation between IEDs and seizure occurrence. Sleep depth was measured by LDP (quantity of 1- to 4-Hz activity in 30-s epochs), and records were scored visually for sleep staging and for IEDs. Multivariate logistic regression analyses were applied. RESULTS: Adjusting for LDP, number of seizures before the current seizure, quartile of the night, and total number of IEDs that occurred during the night, IED did not increase or decrease before seizures (p > 0.1). The rate of IEDs increased directly with LDP (p=0.0001), as shown in prior work. CONCLUSIONS: IEDs are not activated or suppressed before seizures during sleep, suggesting that different pathophysiologic processes underlie these two phenomena. These results corroborate prior studies, while providing a more advanced analysis by adjusting for sleep depth and applying multivariate logistic regression analyses. PMID- 11879387 TI - Prevalence of anti-cardiolipin, anti-beta2 glycoprotein I, and anti-prothrombin antibodies in young patients with epilepsy. AB - PURPOSE: To measure anti-cardiolipin (aCL), anti-beta2 glycoprotein I (anti beta2GPI), and anti-prothrombin (aPT) antibodies in young patients with epilepsy, and to correlate their presence with demographic data, clinical diagnoses, laboratory and neuroradiologic findings, and antiepileptic drugs (AEDs). METHODS: Sera from one hundred forty-two consecutive patients with epilepsy with a median age of 10 years were tested for aCL and anti-beta2GPI autoantibodies by solid phase assays. aPT antibodies also were assayed in sera from 90 patients. Positive results were confirmed after a minimum of 6 weeks. Antinuclear antibodies (ANAs) and antibodies against extractable nuclear antigens (ENAs) also were tested. RESULTS: An overall positivity of 41 (28.8%) of 142 sera was found. Fifteen patients were positive for aCL, 25 for anti-beta2GPI, and 18 for aPT antibodies. Several patients (12%) displayed more than one specificity in their serum. Only one of these patients had a concurrent positivity for ANAs and ENAs. A predominance of younger patients was found in the antibody-positive group. All types of epilepsy were represented in the positive group. No relation between antibody positivity and AEDs was found. Diffuse ischemic lesions at computed tomography (CT)/magnetic resonance imaging (MRI) scans were present in higher percentages in patients who were antibody positive. No positive patient had a history of previous thrombosis or other features related to systemic lupus erythematosus (SLE), and no patient was born of a mother with SLE. CONCLUSIONS: Our study suggests a relation between epilepsy and aPL in young patients. A pathogenetic role for these autoantibodies cannot be excluded, and their determination might prove useful even from a therapeutic point of view. PMID- 11879389 TI - Comparison of statistical parametric mapping and SPECT difference imaging in patients with temporal lobe epilepsy. AB - PURPOSE: Statistical parametric mapping (SPM) is an image-analysis tool that assesses the statistical significance of cerebral blood flow (CBF) changes on a voxel-by-voxel basis, thereby removing the subjectivity inherent in conventional region-of-interest (ROI) analysis. Our platform of single-photon emission computed tomography (SPECT) ictal-interictal difference imaging in clinical epilepsy has been validated for localizing seizure onset. We extend the tools of SPM by further applying statistical measures for the significance of perfusion changes in individual patients to localize epileptogenic foci in patients with defined temporal lobe epilepsy by using paired scans in this preliminary study. METHODS: Twelve patients with pairs of periictal and interictal SPECT scans were analyzed in this comparison study between SPECT difference imaging and SPM difference analysis by using a reference database of paired normal healthy images. These 12 patients possessed seizure foci localized to the mesial temporal lobe as confirmed by surgical outcome and by hippocampal sclerosis on pathology. SPM was used to identify clusters of increased or decreased CBF in each patient in contrast to our control group. RESULTS: The regions having the most significant increased or decreased CBF by SPM analysis were in agreement with regions identified by conventional difference imaging and visual analysis by viewers blinded to the results of the SPM analysis. Differentiated further by time of radiopharmaceutical injection, six of seven patients injected within 100 s of seizure onset displayed hyperperfusion changes localized to the corresponding epileptogenic temporal lobe by both techniques. Among patients receiving injections after 100 s, both techniques showed primarily regions of hypoperfusion, which again were similar between these two methods. CONCLUSIONS: The results provide strong evidence supporting SPM difference analysis in assessing regions of significant CBF change from baseline in concordance with our current clinically used technique of SPECT ictal--interictal difference imaging in epilepsy patients. Difference analysis using SPM could serve as a useful diagnostic tool in the evaluation of seizure focus in temporal lobe epilepsy. PMID- 11879388 TI - Four new families with autosomal dominant partial epilepsy with auditory features: clinical description and linkage to chromosome 10q24. AB - PURPOSE: Autosomal dominant partial epilepsy with auditory features (ADPEAF) is a rare form of nonprogressive lateral temporal lobe epilepsy characterized by partial seizures with auditory disturbances. The gene predisposing to this syndrome was localized to a 10-cM region on chromosome 10q24. We assessed clinical features and linkage evidence in four newly ascertained families with ADPEAF, to refine the clinical phenotype and confirm the genetic localization. METHODS: We genotyped 41 individuals at seven microsatellite markers spanning the previously defined 10-cM minimal genetic region. We conducted two-point linkage analysis with the ANALYZE computer package, and multipoint parametric and nonparametric linkage analyses as implemented in GENEHUNTER2. RESULTS: In the four families, the number of individuals with idiopathic epilepsy ranged from three to nine. Epilepsy was focal in all of those with idiopathic epilepsy who could be classified. The proportion with auditory symptoms ranged from 67 to 100%. Other ictal symptoms also were reported; of these, sensory symptoms were most common. Linkage analysis showed a maximum 2-point LOD score of 1.86 at (theta=0.0 for marker D10S603, and a maximum multipoint LOD score of 2.93. CONCLUSIONS: These findings provide strong confirmation of linkage of a gene causing ADPEAF to chromosome 10q24. The results suggest that the susceptibility gene has a differential effect on the lateral temporal lobe, thereby producing the characteristic clinical features described here. Molecular studies aimed at the identification of the causative gene are underway. PMID- 11879391 TI - Effects of educational intervention on changing parental practices for recurrent febrile convulsions in Taiwan. AB - PURPOSE: To evaluate the effects of educational interventions on parental practices for recurrent febrile convulsions (FC). METHODS: A 2-year follow-up, nonequivalent comparison group design was used to evaluate the intervention effects. Two educational interventions were provided for FC parents in southern Taiwan. The 326 parents voluntarily chose either to receive a mailed pamphlet (n=196) or to attend a 2-h educational program (n=130). Five telephone interviews focused on investigating FC episodes and parental practices for seizures were conducted at months 3, 6, 12, 18, and 24 after the interventions. RESULTS: Of the 326 FC children, 78 (23.9%) had recurrent FCs within the 2-year follow-up. Parents who only received pamphlets did not show significant improvements. Parents who attended the educational program demonstrated significant improvements in the recommended practices, particularly in protecting the convulsing child (8.3 vs. 36.1%; p=0.02 by McNemar) and placing the child on his or her side (19.4 vs. 47.2%; p=0.01). Nonrecommended practices including rushing the convulsing child to the hospital (88.9 vs. 30.6%; p < 0.01) and putting protective devices in the child's mouth (38.9 vs. 8.3%; p < 0.01) significantly decreased. By generalized estimating equation analyses, the types of interventions are the single significant factor influencing parental practice changes from initial to recurrent FCs. CONCLUSIONS: Most parents used inappropriate practices for their child's initial FC. Compared with the mailed pamphlet, the educational program had significant improvements in recommended and nonrecommended practices from initial to recurrent FCs. PMID- 11879390 TI - Temporal lobe epilepsy in children: etiology in a cohort with new-onset seizures. AB - PURPOSE: The aim of this study was to characterize the incidence and etiology of temporal lobe epilepsy (TLE) in a community-based cohort of children with new onset disease. METHODS: A community-based cohort of 30 children with TLE was studied. The patients had new-onset disease before age 14 years between 1995 and 1999. They underwent clinical, EEG, and magnetic resonance imaging investigations. RESULTS: The patients could be divided in three main groups according to likely etiology, as suggested by Harvey et al. (Neurology 1997;49:960-8). Group 1 consisted of eight (26.7%) children with malformations or long-standing, nonprogressive tumors (developmental TLE). Arachnoid cysts were found in three, dual pathology [cortical dysplasia and hippocampal sclerosis (HS)] in one, and focal cortical dysplasia with glioproliferative changes in one patient. Dysembryoplastic neuroepithelial tumor was responsible for the epilepsy in one, and ganglioglioma, in two children. Group 2 consisted of seven (23.3%) children with a significant antecedent and/or HS. Five children had a significant illness or event before the onset of TLE, including perinatal hypoxic-ischemic encephalopathy in one, encephalitis in one, traumatic brain injury in two, and complex febrile seizures in one. HS was found in the patients with traumatic brain injury and complex febrile seizures in the history in addition to two children without known antecedents. Group 3 comprised 15 (50%) children with no abnormality on neuroimaging and no significant antecedents (cryptogenic TLE). CONCLUSIONS: Etiologic differences between children with new-onset TLE may have prognostic implications: children with TLE and significant antecedents/HS are expected to have the greatest risk of continued seizures and psychological problems. PMID- 11879392 TI - Memory outcome after selective amygdalohippocampectomy: a study in 140 patients with temporal lobe epilepsy. AB - PURPOSE: The technique of selective amygdalohippocampectomy (SAH) was originally developed in epilepsy surgery to spare unaffected brain tissue from surgery, thus minimizing the cognitive consequences of temporal lobe surgery. The results of previous studies, however, are equivocal in this regard. This study evaluated memory after SAH in a large sample of patients with mesial temporal lobe epilepsy. METHODS: The 140 patients received material-specific memory tests before and 3 months after unilateral SAH. RESULTS: Significant declines in all aspects of verbal learning and memory were found particularly for the left resected group. With reliability-of-change indices, a high number of patients showed postoperative verbal memory declines, < or = 51% in left SAH and < or = 32% in right SAH. For left SAH, a higher preoperative verbal memory performance, a lower preoperative nonverbal memory score, an older age at surgery, and a later onset of epilepsy predicted a stronger decline in verbal memory. After right SAH, the risk for a verbal memory decline was slightly increased when patients had surgical complications or a presurgical evaluation with bilateral intrahippocampal depth electrodes. Results concerning nonverbal memory were less clear. CONCLUSIONS: The results clearly indicate, that particularly left SAH can lead to a significant decline in memory functions. Predictors of postoperative verbal memory were similar to those reported for temporal lobectomy. Postoperative deteriorations were broader and stronger in our study than in previous studies. We discuss methodologic differences (sample size, retest interval, extent of resection) and other factors as possible reasons. PMID- 11879393 TI - Direct medical costs of refractory epilepsy incurred by three different treatment modalities: a prospective assessment. AB - PURPOSE: More than 20% of epilepsy patients have refractory seizures. Treatment options for these patients include continued polytherapy with/without novel antiepileptic drugs (AEDs), epilepsy surgery (ES), or vagus nerve stimulation (VNS). The purpose of this study was prospectively to compare epilepsy-related direct medical costs (ERDMCs) incurred by these different treatment modalities. METHODS: Eighty-four patients underwent a complete presurgical evaluation protocol at our institution. As a result, 24 (29%) patients were treated with continued AED polytherapy only; 35 (40%) underwent ES; and 25 (30%) had VNS. In each patient, annual costs in the 2 years preceding the therapeutic decision (ERDMC-pre) and during the follow-up afterward (ERDMC-post) were prospectively calculated. Furthermore, frequency of complex partial seizures with/without secondary generalization (CPS+/-SG), dosage and number of AEDs, number of hospital admission days, clinic visits, and laboratory tests before and after the therapeutic decision also were prospectively assessed. ERDMC-pre and ERDMC-post were compared in and among the three treatment groups. RESULTS: In patients conservatively treated with AEDs, mean frequency of CPSs decreased from 12 per month to nine per month, whereas mean ERDMCs decreased from $2,525 U.S. to $2,421 U.S. In surgical patients, mean seizure frequency decreased from six to fewer than one per month; mean ERDMCs per year decreased from $1,465 U.S. preoperatively to $1,186 U.S. postoperatively. In the VNS group, mean seizure frequency decreased from 21 per month to seven per month. ERDMCs in this subgroup decreased from $4,826 U.S. to $2,496 U.S. Mean seizure frequency changes were significant when conservatively treated patients were compared with surgically treated and VNS patient groups (chi2 test, p<0.001 and p=0.0019, respectively). ERDMC changes in conservatively treated patients also were statistically significant when compared with surgically treated and VNS patients (chi2 test, p=0.0007 and p=0.0036, respectively). No statistically significant differences were found in ERDMC changes between the surgical and VNS groups (chi2 test, p=0.387). CONCLUSIONS: Ongoing daily treatment of patients who underwent resective surgery costs significantly less than conservative treatment. For patients in whom resective surgery is not an option, ERDMC show a significant decrease in VNS-treated patients compared with conservatively treated patients. PMID- 11879394 TI - Diet enriched with omega-3 fatty acids alleviates convulsion symptoms in epilepsy patients. AB - PURPOSE: We examined whether a dietary supplement containing omega-3 polyunsaturated fatty acids (n-3 PUFAs) can alleviate and/or reduce the frequency of epileptic seizures in patients with central nervous system (CNS) diseases treated with anticonvulsive drugs (ACDs). METHODS: A special spread containing 65% n-3 PUFAs was added to the daily diet. The patients consumed 5 g of this spread at every breakfast for 6 months. RESULTS: Five patients completed the study. In all of them, a marked reduction in both frequency and strength of the epileptic seizures was recorded. CONCLUSIONS: Incorporation of the dietary supplement containing n-3 PUFAs may be beneficial in suppression of some cases of epileptic seizures. PMID- 11879396 TI - Keeping up appearances. PMID- 11879395 TI - The Henri GASTAUT International Award of the French League. PMID- 11879399 TI - Abstracts for BSPD Awards. PMID- 11879398 TI - Editorial. PMID- 11879400 TI - Editorial. PMID- 11879411 TI - Congres IAPD-SFOP Paris 2001. PMID- 11879409 TI - Editorial. PMID- 11879412 TI - The United Kingdom research assessment exercise (RAE) 2001. PMID- 11879414 TI - Getting your paper to the right journal: a case study of an academic paper. AB - BACKGROUND: The scientific community views the publication of academic papers as a means of disseminating information, ensuring transparency and good practice in terms of research utilization. However, the choice of journal in which to publish is frequently influenced by other, less obvious, factors. This paper describes the lengthy route taken to get a methodological paper about pilot studies into print. AIM AND METHOD: This paper shares some of our experiences and discusses the lessons that we learned about the process of getting into print. A case study approach is adopted to help the reader understand the different influences on this process. FINDINGS: Our methodological paper was submitted to six different academic journals before it was finally accepted for publication. The choice of journal was influenced by the need to reach an appropriate academic audience, the estimated turn around time (the time taken between submission of a paper and its subsequent publication) and the level of academic credibility of the journal (often assessed by the journal's Impact Factor). Publishing in 'high impact' academic journals assumed considerable importance for us in view of the UK Research Assessment Exercise (RAE). However, the consequence of going down the journal status hierarchy was that we lost about 1 year through submitting, rewriting according to each of the journal's specific requirements and resubmitting. The case study also demonstrates that getting into print often depends on a range of factors, not just the quality of the written text. CONCLUSIONS: Getting a paper published may depend not only on the intrinsic quality of the paper, but also whether it is submitted to the 'right' academic journal. Moreover, if journals do not take certain papers (e.g. ones with negative findings or those reporting multi-disciplinary studies) then this can lead to publication bias. PMID- 11879415 TI - Feminist poststructuralism: a methodological paradigm for examining clinical decision-making. AB - AIM: To present the philosophical framework of feminist poststructuralism, discuss its use as an innovative research approach and its implications for nursing knowledge development and practice. BACKGROUND: This perspective examines the construction of meaning, power relationships, and the importance of language as it affects contemporary healthcare decisions. It seeks to identify and expose biases that marginalize the healthcare needs of women and contribute to healthcare disparities for this population. Additionally, a feminist poststructuralist perspective seeks to develop new knowledge for understanding gender differences. NURSING APPLICATION: A feminist poststructuralist perspective represents an alternative paradigm for studying the phenomenon of clinical decision-making. An empirical application example of a feminist poststructuralist perspective is provided. This exemplar investigated emergency department registered nurses' triage decisions for men and women with symptoms suggestive of coronary heart disease. PMID- 11879416 TI - Assessing the skills for family planning nurse prescribing: development of a psychometrically sound training needs analysis instrument. AB - RATIONALE AND AIMS: Family planning nurses have been identified for early development for prescribing authority in the United Kingdom (UK). Currently, no psychometrically founded training needs analysis instrument exists that can reliably assess the nature and extent of the specific educational provision required for this role. This paper is concerned with the development of an instrument capable of defining the development needs for family planning nurse prescribing. METHODS: A national survey was conducted with 388 family planning nurses, using a modified training needs analysis instrument. Respondents were required to assess the importance of 40 tasks, firstly for the role of the family planning nurse (FPN), and again for the role of the family planning nurse prescriber (FPNP). The data from each set of ratings were separately factor analysed using orthogonal Varimax rotations and Cronbach's alpha was computed for each factor. RESULTS: Six factors emerged from the family planning nurse ratings ('professional development', 'managing patient consultations', 'critical appraisal', 'clinical information giving and professional accountability', 'collaborative working and current National Health Service (NHS) issues' and 'dispensing of drugs') and nine factors emerged from the FPNP ratings ('research and practice development', 'prescribing and professional accountability', 'management/leadership', 'clinical decision making and risk assessment', 'advanced health assessment', 'critical appraisal', 'referral processes', 'core nursing skills' and 'dispensing of drugs'. This suggests that the role of the FPNP is more extended and, moreover, that the two roles are configured very differently but in a way that makes logical and coherent sense within existing research and government policy. This indicates that the instrument is valid. Moreover, all but two of the factors had a Cronbach's alpha score of >0.7 and so can be considered reliable. CONCLUSIONS: The results indicate that the modified instrument is valid and reliable and therefore can be used with confidence to assess the training needs of FPNPs. In addition, the factors have outlined a cogent definition of the role of the FPNP, which can be used both to inform educational programmes and to assess their efficacy. PMID- 11879417 TI - Development of an instrument to measure strategic and clinical quality indicators in postoperative pain management. AB - AIM OF THE STUDY: To develop and evaluate psychometric properties, that is reliability and validity, of an instrument to measure strategic and clinical quality indicators in postoperative pain management. BACKGROUND: Strategic and clinical quality indicators in postoperative pain management were previously developed from a tentative model of important aspects of surgical nursing care and assessed to have content validity, that is to be essential for the quality of care, realistic to carry out and possible for nurses to use to influence management. METHODS: The quality indicators were converted to items suitable for a patient questionnaire and were scored on a 5-point scale, with higher scores indicating higher quality of care. Inpatients from five surgical wards took part in this study on their second postoperative day. The response rate was 96% and the average ages of the female (n=120) and the male (n=78) respondents were 62 and 63 years, respectively. RESULTS: Items in the total scale had an average inter-item correlation >0.20 and an item-total correlation >0.30. Cronbach's coefficient alpha was 0.84 for the total scale. Four factors entitled 'communication', 'action', 'trust' and 'environment' emerged from an orthogonal factor analysis, with a cumulative variance of 61.4%. Patients who received epidural analgesia had higher scores on the total scale compared with those who did not receive epidural analgesia. Patients who reported more pain than expected had lower scores on the total scale compared with those who did not report more pain than expected. Correlation between the total scale and an overall pain relief satisfaction question was 0.53. CONCLUSION: The results suggest initial support for the new instrument as a measure of strategic and clinical quality indicators in postoperative pain management, but it must be further refined, tested and evaluated. PMID- 11879418 TI - The cooling-suit: case studies of its influence on fatigue among eight individuals with multiple sclerosis. AB - AIM OF THE STUDY: To study if the use of a cooling-suit by individuals with multiple sclerosis (MS) influenced their experience of fatigue and consequent restrictions in daily life. BACKGROUND: The majority of MS patients consider fatigue as one of their most disabling symptoms and as having a significant impact on their daily lives. Fatigue often increases in a warm environment. A cooling-suit has been reported as a practical method of cooling, but the effect on fatigue has not yet been studied. RESEARCH METHODS: Eight individuals used a cooling-suit in their own homes during a test-period. In a single case-control design, their experience of fatigue was studied before and after. Quantitative and qualitative methods were used: self-assessments using the Fatigue Impact Scale (FIS), open-ended interviews and semi-structured diaries. RESULTS: All study participants reported a reduction in fatigue during the test period. On the FIS, they scored reductions in the physical, cognitive and psycho-social dimensions of daily life. They experienced fatigue less often and for shorter periods. In their diaries and interviews they described decreased muscular strain, less sense of fatigue in relation to intake of food and positive effects on cognitive, social or affective problems related to fatigue. DISCUSSION: The cooling-suit is a practical method for cooling. It gives freedom and flexibility and can be used regardless of setting. Nurses who meet heat-sensitive individuals with MS have the opportunity to give information on cooling methods, including how to use a cooling-suit. In this pilot study we found that individuals with MS who suffered from fatigue reported a number of improvements in quality of daily life. CONCLUSIONS: The result indicates that use of a cooling-suit by individuals with MS may decrease their sense of fatigue. In this sample positive outcomes on daily life situations were reported. Further studies are needed to support these results. PMID- 11879419 TI - Caring for maltreated children: a challenge for health care education. AB - AIM OF THE STUDY: The aim of this study was to establish whether paediatric nurses and doctors in a university hospital need supplementary training in the identification of child physical abuse and whether there is a need to develop health care education on child abuse issues. BACKGROUND: Child physical abuse was defined as actions of a parent or a primary caregiver of a child under 18 years of age, which have caused physical injuries to the child. METHODS: The sample consisted of paediatric staff (n=513) in a university hospital. Data were collected with a questionnaire, and the response rate was 62%. The data were analysed using statistical methods and quantitative content analysis. FINDINGS: Sixty per cent of respondents needed supplementary training in the identification of child abuse. Those having experience in caring for abused children considered the identification most difficult (P < 0.001) and needed training more often (P < 0.005) than the others. The need for training was explained by stating that their basic education had not addressed child abuse issues, and by the complexity of the issue. CONCLUSIONS: The findings indicate that research-based knowledge of the identification and treatment of child abuse should be incorporated into health care education. Supplementary education should address the management of complex and sensitive issues with customers, legislation, division of labour and multiprofessional collaboration in which nurses and doctors have an important, family wellbeing promoting role. PMID- 11879420 TI - Aspects of curriculum policy in preregistration nursing education in the Republic of Ireland: issues and reflections. AB - AIMS: This paper sets out to examine critically aspects of curriculum policy related to preregistration nursing education in the Republic of Ireland. BACKGROUND: Following a period of industrial unrest concerned with pay, promotional opportunities and conditions of employment amongst nurses and midwives in the Republic of Ireland, a Commission on Nursing was established in March 1997. Along with a series on recommendations pertaining to the role and the professional development of nurses and midwives, the final report of the Commission on Nursing contained recommendations on the educational preparation of nurses. Specifically, the Commission recommended that the future framework for preregistration nursing education in the Republic of Ireland be based on a 4-year degree programme, fully integrated into higher education. As a means of giving effect to this recommendation, the Commission also recommended the establishment of a Nursing Education Forum, whose task it was to develop a strategic framework for the introduction of the proposed preregistration nursing degree programme. The Forum published its final report in October 2000 and this report constituted the most explicit statement of official policy on the nursing curriculum in the Republic of Ireland. DESIGN: The paper constitutes a critical analysis of selected aspects of the report of the Nursing Education Forum, in the light of scholarly literature on curriculum philosophy and curriculum policy. SUMMARY OF CONTENT: The paper considers some of the implications for curriculum development and design arising out of the recommendations of the Nursing Education Forum. Among a list of core principles underpinning curriculum regulation and design, the report included the principle of 'eclecticism'. The paper considers this principle with reference to nursing epistemology, pedagogical practice and curriculum policy, and seeks to challenge some of the assumptions underlying this principle. CONCLUSIONS: The epistemological identity and the structural integrity of primary forms of knowledge in the preregistration curriculum need to be maintained if students are to develop their knowledge and understanding of nursing science. PMID- 11879421 TI - Student--teacher connection in clinical nursing education. AB - BACKGROUND: The trend toward humanistic nursing education has called for a transformed student-teacher relationship that fosters learning and growth of students and teachers. Although such a relationship has been claimed to form the basis for student-teacher connection and to be a positive influence on students' learning outcomes, there is a paucity of research exploring these claims. Neither the nature of student-teacher connection nor the processes by which it occurs have been described. AIMS: A research study was undertaken to explore and describe undergraduate nursing students' experiences of connection within the student-teacher relationship and the effects of student-teacher connection on students' learning experiences in clinical nursing education. RESEARCH DESIGN: The qualitative research approach of interpretive description was chosen for this study. Unstructured interviews and a focus group were used to collect data from eight undergraduate nursing students. Data were analysed using the process of constant comparative analysis, and revealed four interrelated major categories that formed a description of the students' experience of student-teacher connection. FINDINGS: This article presents part of the findings of this study. After describing the nature of student-teacher connection, the discussion focuses on the influence of teachers and other factors on the formation of student teacher connection. Relevance is given to this discussion by describing the outcomes of connection for students' clinical learning experiences. PMID- 11879422 TI - Getting evidence into practice: the role and function of facilitation. AB - AIM OF PAPER: This paper presents the findings of a concept analysis of facilitation in relation to successful implementation of evidence into practice. BACKGROUND: In 1998, we presented a conceptual framework that represented the interplay and interdependence of the many factors influencing the uptake of evidence into practice. One of the three elements of the framework was facilitation, alongside the nature of evidence and context. It was proposed that facilitators had a key role in helping individuals and teams understand what they needed to change and how they needed to change it. As part of the on-going development and refinement of the framework, the elements within it have undergone a concept analysis in order to provide theoretical and conceptual clarity. METHODS: The concept analysis approach was used as a framework to review critically the research literature and seminal texts in order to establish the conceptual clarity and maturity of facilitation in relation to its role in the implementation of evidence-based practice. FINDINGS: The concept of facilitation is partially developed and in need of delineation and comparison. Here, the purpose, role and skills and attributes of facilitators are explored in order to try and make distinctions between this role and other change agent roles such as educational outreach workers, academic detailers and opinion leaders. CONCLUSIONS: We propose that facilitation can be represented as a set of continua, with the purpose of facilitation ranging from a discrete task-focused activity to a more holistic process of enabling individuals, teams and organizations to change. A number of defining characteristics of facilitation are proposed. However, further research to clarify and evaluate different models of facilitation is required. PMID- 11879423 TI - Spirituality and health: towards a framework for exploring the relationship between spirituality and health. AB - BACKGROUND: A growing body of evidence has found that spirituality enhances health. However, spirituality is an elusive concept that defies clear definition. This inevitably presents difficulties when comparing the findings of studies. Therefore conceptual clarification is essential if practitioners are to better understand the relationship between spirituality and health. AIMS: The aim of this paper is to develop a conceptual framework, which can be used to explore the relationship between spirituality and health. METHODS: The concept-indicator model was used to analyse spirituality in the literature. The literature was searched for empirical indicators or what are taken as essential attributes of spirituality. Similarities and differences between approaches were identified and these formed the basis of a framework. FINDINGS: The analysis identified three approaches (a trichotomy) to spirituality in the literature. These were termed the transcendent, the value guidance and the structuralist-behaviourist approaches. The paper shows how by clarifying the different conceptualizations of spirituality and the interrelationship between them researchers can also clarify their respective contributions to health. Thus a contribution is made towards making more explicit the ways in which key aspects of spirituality such as transcendence, meaning and purpose, connectedness, hope, and faith, work to produce health benefits in terms of prevention, recovery from illness, or coping with illness. CONCLUSIONS: The framework (or trichotomy) will enable practitioners to understand better the connection between spirituality and health. In particular, it will show that to appreciate the benefits that patients might experience from their value or belief systems, practitioners must actively explore the content of those systems in a respectful way. PMID- 11879424 TI - An analysis of the concept of equity and its application to health visiting. AB - AIMS: The aim of this paper is to provide an in-depth analysis of the concept of equity in the British health visiting service and thus facilitate understanding of the term. An attempt has been made to apply the analytical process to the context of health visiting. BACKGROUND: Equity has been discussed at some length in the health literature. Much of this including the health visiting literature discusses inequality and inequalities in health at the same time as discussing equity. Increasingly, poverty and inequalities have become accepted as determinants in poor health. It seemed appropriate therefore to analyse these topical and seemingly related concepts. METHODS: A well established method of concept analysis has been utilized to facilitate the process of analysis. RESULTS: Confusion between the meaning and application of the concepts of equity and equality was found. Much of the health visiting and health care literature related to addressing the needs of groups who experience inequality in terms of service provision, and access to services. Following an in-depth critical analysis of the literature three critical attributes of the concept were identified: equal opportunity of access to services; a high standard of service for everyone; and unequal distribution of services to meet unequal need. These relate to two distinct forms of equity: horizontal and vertical equity. A definition of equity has been proposed. Maxwell's (1992) framework is suggested as a means to evaluate equity in service provision and uptake. CONCLUSIONS: Most of the literature examines equity at a macro level, that is service level. Nurses and health visitors are urged to examine the services that they are involved in to determine the level of equity that is achieved or not attained. Suggestions have been offered as to how this task may be undertaken. A definition has been proposed to stimulate debate and discussion about the concept within nursing contexts. PMID- 11879425 TI - Freestanding lipid bilayers as substrates for electron cryomicroscopy of integral membrane proteins. AB - Phospholipid bilayers, 40 A thick, were generated as electron microscope substrates by submerging copper grids overlaid with holey plastic through a lipid monolayer on a water surface. Previously formed proteoliposomes containing single particle membrane proteins in their bilayers were then fused into the newly formed bilayer substrate. To demonstrate this methodology, multi-drug resistance protein P-glycoprotein was incorporated into these bilayers and imaged by fixed beam microscopy and scanning transmission electron microscopy. PMID- 11879426 TI - A technique for the examination of polar ice using the scanning electron microscope. AB - The microstructure and location of impurities in polar ice are of great relevance to ice core studies. We describe a reliable method to examine ice in the scanning electron microscope (SEM). Specimens were cut in a cold room and could have their surfaces altered by sublimation either before (pre-etching) or after (etching) introduction to the cryo-chamber of the SEM. Pre-etching was used to smooth surfaces, whilst etching stripped away layers from the specimen surface, aiding the location of particles in situ, and allowing embedded structures to be revealed. X-ray analysis was used to determine the composition of localized impurities, which in some cases had been concentrated on the surface by etching. Examining uncoated surfaces was found to be advantageous and did not detract from qualitative X-ray analysis. Imaging uncoated was performed at low accelerating voltages and probe currents to avoid problems of surface charging. PMID- 11879428 TI - Phase evolution in cholesterol/DPPC monolayers: atomic force microscopy and near field scanning optical microscopy studies. AB - A combination of atomic force microscopy (AFM) and near field scanning optical microscopy has been used to study domain formation in dipalmitoylphosphatidylcholine (DPPC)/cholesterol monolayers with cholesterol concentrations ranging from 0 to 50%. The results show a clear evolution from a mixture of liquid expanded and liquid condensed phases for cholesterol concentrations < 10% to a mixture of liquid expanded and two cholesterol containing phases at intermediate concentrations, and finally to a single homogeneous liquid ordered phase for 33% cholesterol. Mixtures of the various phases are clearly identified by height differences in AFM and in some cases by fluorescence imaging for samples containing 0.5% BODIPY dye, which localizes preferentially in the more fluid phase. Note that fluorescence imaging, at least with the dye used here, is unable to distinguish between the cholesterol-rich and cholesterol-poor phases detected at intermediate cholesterol concentrations. The combination of fluorescence and AFM imaging provides a more complete picture of the phase evolution for cholesterol/DPPC monolayers than could be obtained by either technique alone, and presents substantial advantages over conventional fluorescence microscopy in that submicrometre-sized domains can be readily detected. PMID- 11879427 TI - A method for quantifying cell size from differential interference contrast images: validation and application to osmotically stressed chondrocytes. AB - An automatic image analysis method was developed to determine the shape and size of spheroidal cells from a time series of differential interference contrast (DIC) images. The program incorporates an edge detection algorithm and dynamic programming for edge linking. To assess the accuracy and working range of the method, results from DIC images of different focal planes and resolutions were compared to confocal images in which the cell membrane was fluorescently labelled. The results indicate that a 1-microm focal drift from the in-focus plane can lead to an overestimation of cell volume up to 14.1%, mostly due to shadowing effects of DIC microscopy. DIC images allow for accurate measurements when the focal plane lies in a zone slightly above the centre of a spherical cell. In this range the method performs with 1.9% overall volume error without taking into account the error introduced by the representation of the cell as a sphere. As a test case, the method was applied to quantify volume changes due to acute changes of osmotic stress. PMID- 11879429 TI - Influence of metal roughness on the near-field generated by an aperture/apertureless probe. AB - We study the influence of metal roughness on the near-field distribution generated by an aperture or an apertureless (scattering) probe. Different experimental parameters are investigated: roughness magnitude, aperture form, distribution of the roughness. Our results show that aluminium roughness has a dramatic impact on the emission characteristics of a near-field probe and in particular on its polarization sensitivity. Apertureless or scattering probes appear to be less sensitive to roughness and to provide a well confined field even with a somewhat rough probe. PMID- 11879430 TI - Relative labelling index: a novel stereological approach to test for non-random immunogold labelling of organelles and membranes on transmission electron microscopy thin sections. AB - Simple and efficient protocols for quantifying immunogold labelling of antigens localized in different cellular compartments (organelles or membranes) and statistically evaluating resulting labelling distributions are presented. Two key questions are addressed: (a) is compartmental labelling within an experimental group (e.g. control or treated) consistent with a random distribution? and (b) do labelling patterns vary between groups (e.g. control vs. treated)? Protocols rely on random sampling of cells and compartments. Numbers of gold particles lying on specified organelle compartments provide an observed frequency distribution. By superimposing test-point lattices on cell profiles, design-based stereology is used to determine numbers of points lying on those same compartments. Random points hit compartments with probabilities determined by their relative sizes and so provide a convenient internal standard, namely, the expected distribution if labelling is purely random. By applying test-line lattices, and counting sites at which these intersect membrane traces, analogous procedures provide observed and expected labelling distributions for different classes of membranes. Dividing observed golds by expected golds provides a relative labelling index (RLI) for each compartment and, for random labelling, the predicted RLI = 1. In contrast to labelling densities of organelles (golds microm(-2) or membranes (golds microm( 1)), RLI values are estimated without needing to know lattice constants (area per point or length per intersection) or specimen magnification. Gold distributions within a group are compared by chi-squared analysis to test if the observed distribution differs significantly from random and, if it is non-random, to identify compartments which are preferentially labelled (RLI > 1). Contingency table analysis allows labelling distributions in different groups of cells to be compared. Protocols are described and illustrated using worked specimen examples and real data. PMID- 11879431 TI - New approach to optical diffraction tomography yielding a vector equation of diffraction tomography and a novel tomographic microscope. AB - We first obtain a frequency-space equation of diffraction tomography for the electric field vector, within the first-order Born approximation, using a simplified formalism resulting from using three-dimensional spatial frequencies and replacing outgoing waves by linear combinations of homogeneous plane waves. A coherent optical diffraction tomographic microscope is then described, in which a sample is successively illuminated by a series of plane waves having different directions, each scattered wave is recorded by phase-shifting interferometry, and the object is then reconstructed from these recorded waves. The measurement process in this device is analysed taking into account the illuminating wave, the wave scattered by the sample, the reference wave, and the phase relations between these waves. This analysis yields appropriate equations that take into account the characteristics of the reference wave and compensate random phase shifts. It makes it possible to obtain a high-resolution three-dimensional frequency representation in full conformity with theory. The experimentally obtained representations show index and absorptivity with a resolution limit of about a quarter of a wavelength, and have a depth of field of about 40 microm. PMID- 11879432 TI - Counting dendritic spines in brain tissue slices by image correlation spectroscopy analysis. AB - Growth of new micrometre sized projections called dendritic spines in neurones has been linked to the encoding of long-term memories in vertebrates. Numerous studies have been carried out at both the light and electron microscopy level to quantify dendritic spine densities in brain tissue in laboratory animals. Currently, such efforts using light microscopy have relied on manual counting of spines in confocal or two-photon optical slice images of tissue containing fluorescently labelled spines. This manual approach can be slow and tedious, especially for samples with high spine densities. We introduce an alternative way of performing spine counting that uses an applied image intensity threshold followed by spatial image correlation spectroscopy (ICS) analysis. We investigated the effect of particle sizes above the diffraction limit on the autocorrelation analysis as well as the influence of background fluorescence. Our results show that, for well labelled cerebellar tissue samples imaged with a signal-to-noise ratio of 5 or greater, ICS-based spine counts can be conducted with the same 15-20% precision as manual counting, but much more rapidly. PMID- 11879434 TI - 3D in-vivo X-ray microtomography of living snails. AB - In this paper we report the first in-vivo scanning of living snails by desktop X ray microtomograph with a resolution up to 10 m. Consecutive cross-sections were acquired without destroying the specimen. Subsequently, 3D images were reconstructed. The results clearly demonstrate the possibilities of in-vivo scanning. Processes of growth and regeneration of living snails were visualized over a period of time. PMID- 11879433 TI - Automated high-throughput electron tomography by pre-calibration of image shifts. AB - Electron tomography is a versatile method for obtaining three-dimensional (3D) images with transmission electron microscopy. The technique is suitable to investigate cell organelles and tissue sections (100-500 nm thick) with 4-20 nm resolution. 3D reconstructions are obtained by processing a series of images acquired with the samples tilted over different angles. While tilting the sample, image shifts and defocus changes of several microm can occur. The current generation of automated acquisition software detects and corrects for these changes with a procedure that incorporates switching the electron optical magnification. We developed a novel method for data collection based on the measurement of shifts prior to data acquisition, which results in a five-fold increase in speed, enabling the acquisition of 151 images in less than 20 min. The method will enhance the quality of a tilt series by minimizing the amount of required focus-change compensation by aligning the optical axis to the tilt axis of the specimen stage. The alignment is achieved by invoking an amount of image shift as deduced from the mathematical model describing the effect of specimen tilt. As examples for application in biological and materials sciences 3D reconstructions of a mitochondrion and a zeolite crystal are presented. PMID- 11879435 TI - A novel sample holder allowing atomic force microscopy on transmission electron microscopy specimen grids: repetitive, direct correlation between AFM and TEM images. AB - A novel sample holder that allows atomic force microscopy (AFM) to be performed on transmission electron microscope (TEM) grids is described. Consequently, AFM and TEM images were repeatedly obtained on exactly the same sample area. For both techniques, a thin carbon film was used as the imaging substrate. Although these techniques have been previously used in conjunction, AFM and TEM images on exactly the same area have not been repeatedly obtained for any system. Correlation of AFM and TEM images is useful for work where the three-dimensional topographical information provided by the AFM could be used to better interpret the two-dimensional images provided by the TEM and vice versa. To demonstrate the applicability of such correlation, new results pertaining to a fibrillar collagen system are summarized. PMID- 11879436 TI - New imaging modes for lenslet-array tandem scanning microscopes. AB - The tandem scanning microscope permits confocal images to be obtained in real time and viewed directly by eye. The light budget of these instruments may be increased from a few percent to a few tens of percent by incorporating an array of microlenses so as to increase the amount of illumination light that reaches the specimen. These instruments are configured for fluorescence imaging together with laser illumination. We describe how the versatility of the instrument may be enhanced to permit the use of incoherent light sources as well as extending the imaging modes to include bright-field reflection. PMID- 11879438 TI - The challenge of balancing different agendas. PMID- 11879439 TI - Medical dominance in multidisciplinary teamwork: a case study of discharge decision-making in a geriatric assessment unit. AB - AIM: To investigate the degree of medical dominance in multidisciplinary teams in a geriatric assessment unit by focusing on decision-making with regard to patient discharge. BACKGROUND: The persistence of medical dominance in multidisciplinary teamwork has been widely assumed but insufficiently researched, particularly through close observation of team practice. The present study seeks to rectify this by examining the extent of medical dominance in two multidisciplinary teams working in a hospital-based geriatric assessment unit. METHODS: Team practice was analysed by observing and audiotaping five case review meetings in each team and by semi-structured interviews with team members. RESULTS: In terms of level of contribution, the issues raised at meetings, and the team responses to discharge initiation, a lower than expected level of medical dominance was identified. This lower than expected level is related to consultants' views on the nature of rehabilitation, leading to a consensus amongst team members as to the purpose of geriatric assessment, and to a high level of team stability. CONCLUSION: Reducing the level of medical dominance encourages the contributions of all team members and thus enhances patient care. More training in team skills would also be beneficial, including interprofessional training. PMID- 11879440 TI - Nurses' use of time in a medical-surgical ward with all-RN staffing. AB - AIMS: To investigate allocation of nursing time, organisation of nursing activities and whether or not allocation and organisation have changed over time. BACKGROUND: In a ward that changed to all-RN staffing, the nurses were encouraged to implement a patient-focused philosophy. The nurses perceived that they had difficulty in using the time available efficiently. METHODS: Non-participant observations were conducted with 2-year intervals. Ten consecutive weekdays were covered on two occasions. The study was carried out at a university hospital in Sweden. FINDINGS: Between observations, a significant change in the organization of the direct care had occurred, and the same tendency was found in patient administration and general management. The organization of work changed from a partly fragmented to a more coherent one. The time used for direct care and administrative activities increased between the two observations, while indirect care, personal and service activities decreased. CONCLUSION: It can be suggested that the nurses used their time efficiently and, over time, they developed a more coherent way of organizing nursing activities. PMID- 11879441 TI - Research utilization among medical and surgical nurses: a comparison of their self reports and perceptions of barriers and facilitators. AB - BACKGROUND: Although the number of studies on research utilization is steadily growing, there are only a few specialty-specific studies. AIMS: This study focuses on medical and surgical nurses. It compares their reported use of research utilization and their perceptions of barriers and obstacles. METHODS: A convenient sample of 210 medical nurses and 269 surgical nurses, from 10 general hospitals in 14 Trusts in Northern Ireland (NI), was surveyed. RESULTS: The results showed that the reported extent of research utilization was high, with less than 10% in each group reporting never/seldom using research. The reported difference between the medical and surgical nurses was very small, with medical nurses reporting a slightly higher rate of utilization. However, this was not statistically significant at 5% level. CONCLUSIONS: The Barriers Scale (Funk et al. 1991a) used in this study to assess their perceptions of barriers and facilitators revealed a similar picture for both groups. The top two barriers were "Management will not allow implementation" and "The nurse does not feel she/he has enough authority to change patient care procedures". These findings, as well as the need to take research utilization studies further, are discussed. PMID- 11879442 TI - The needs of informal carers: a proposed assessment tool for use by public health nurses. AB - AIM: To develop an assessment tool for the collection of information on carers' needs and to pilot test same. BACKGROUND: No formal assessment of the needs of carers is undertaken by Public Health Nurses (PHNs) in the West of Ireland. METHOD: An assessment tool which took the form of a questionnaire was designed based on an earlier needs analysis, a literature review and qualitative data obtained with carers at two focus groups. Sixty carers were involved in the pilot study of the tool. FINDINGS: Carers found this model made them feel valued, created awareness about the effects of caring and provided them with information. They recommended its introduction. CONCLUSION: The carers and PHNs found the assessment tool to be useful and comprehensive in its approach. However, it requires modification, with specific attention being paid to the validity and reliability of the tool. PMID- 11879443 TI - An alternative technique for evaluating the effectiveness of continuing professional development courses for health care professionals: a pilot study with practice nurses. AB - BACKGROUND: Increasing recognition of the need for continuing professional development (CPD) for health professionals has created an inevitable expansion in course provision. One criterion of a successful course demands that CPD must meet local health service needs, as well as the personal and professional requirements of the participants. At a more general level, a key pedagogical requirement of any educational course is that the content and process must meet the stipulated aims and objectives, and this requires appropriate evaluation procedures. Within health care CPD, very little course evaluation routinely takes place, possibly because the area is relatively new, and secondly because relevant evaluation may be difficult to operationalize. AIMS: The current study reports a small exploratory pilot investigation into the use of a valid and reliable training needs analysis questionnaire as a means of evaluating a nurse practitioner (NP) training scheme for practice nurses (PN). METHODS: Using a before/after design, the instrument involved the participants in a pre and post course self-assessment on a range of tasks. These assessments were made according to three criteria: how critical each task is considered to be to the effective delivery of the respondent's current PN role; how critical each task is considered to be to the effective delivery of the role of the NP; and how well each task is currently performed. Various comparisons of the ratings provide a considerable amount of information that may be of use to the nurse manager in planning NP programmes. Of particular relevance to the current paper, however, are the comparisons between salience of tasks for the NP role and performance on these, before and after the course. This provides an assessment of the extent to which the course has met educational requirements. RESULTS: The present study demonstrated that overall, the course reduced skill deficits in all but seven of the tasks. Of these, six related to research and audit and the seventh to clinical examination of patients. CONCLUSION: The study is discussed in terms of the implications of the technique for course development and planning to take account of local needs, as well the instrument's reliability and validity, for use in this way. PMID- 11879444 TI - The staff's satisfaction with the hospital bed. AB - AIM: To determine how nurses experience the functions of hospital beds and the bedside equipment when performing specified common nursing tasks. BACKGROUND: The hospital bed is an important working tool for the nursing staff but few studies have focused on hospital beds at acute wards. METHOD: Data was collected with questionnaires answered by 74 registered and enrolled nurses from three Departments. FINDINGS: The bed was frequently used when performing nursing tasks with patients in bed, but the manoeuvring features were difficult to adjust, resulting in unsatisfactory working postures. Transporting the patient in bed and cleaning of the bed were the most troublesome tasks. The bed clothes were often experienced as unsatisfactory for their patients. CONCLUSIONS: The beds have serious deficiencies. It is imperative to improve them, as a good physical milieu positively influences the working conditions of the staff. PMID- 11879447 TI - Joys, frustrations and concerns of a journal peer reviewer. PMID- 11879448 TI - A study of the criteria used by healthcare professionals, managers and patients to represent and evaluate quality care. AB - AIM: To explore the perceptions of and criteria used by healthcare professionals, managers, patients and relatives to represent and evaluate their concept of quality care. METHODS: A qualitative approach using grounded theory was adopted in this exploratory descriptive study. Data collected by semi-structured interviews from a purposive sample of nurses, doctors, managers (n = 36), patients (n = 34) and relatives (n = 7) from one acute medical ward, were subjected to content, question and thematic analysis, using an inductive categorizing scheme. FINDINGS: Three categories of criteria relating to Care Resources, Processes and Outcomes were identified by healthcare professionals, managers, patients and relatives. Resource criteria included Human Resources: staff numbers, ratio to patients, skill mix; as well as Environmental/Physical and Financial Resources. Process criteria included Care Functions, Practices and Standards as well as Interpersonal Processes. Outcome criteria were either patient-focused: feeling comfort, happy, informed and satisfied; or health related: maintenance or progress with health problems and goals. CONCLUSIONS: The criteria used by healthcare stakeholders in this study were not unusual; virtually all were supported by the literature, a proportion of which was evidence-based. The criteria identified in this study are however consensual, agreed upon by healthcare professionals, managers, patients and relatives as representing their view of quality care. These consensual criteria could be used as unifying constructs for the development and testing of more comprehensive, reliable and valid methods of evaluating quality care which represent its multiple dimensions and perspectives. PMID- 11879450 TI - The role and preparation of first-line nurse managers in Australia: where are we going and how do we get there? AB - Organizational restructuring is impacting heavily on the first-line nurse managers' role. Given their critical role, it is important that the best and most talented are attracted to and prepared to be first-line managers. The authors provide an overview of the role of first-line nurse managers in Australia, proposing that nurse administrators take a more active role than has traditionally been the case in ensuring staff are appropriately educated and selected for management positions. PMID- 11879449 TI - Participation in arranging continuing health care packages: experiences and aspirations of service users. AB - AIMS: This paper examines patients' and carers' experiences of receiving community health services, and considers the degree of patients' participation in the management of their continuing care. BACKGROUND: Care management, advocated for many years as a way of ensuring appropriate and coordinated care, emphasizes the involvement of patients and carers in care planning. Evidence suggests that such involvement is unusual. METHODS: Semi-structured interviews were carried out with 99 continuing health care patients and/or their carers. FINDINGS: A few informants had willingly chosen to be active in arranging and coordinating their care, and a few others, who did not take an active role, were very satisfied with the services which they received. The majority, however, were not satisfied, feeling unclear about how their needs had been assessed and how services had been arranged. They were mostly disappointed by the absence of information and by the lack of regular contact with NHS and/or Social Services personnel. Some had felt compelled to become proactive in order to ensure that the care provided was adequate. CONCLUSIONS: Community nurses are valued by patients, and are well placed to be care managers. However, less qualified staff are also able to offer the regular support and information which patients want and value. PMID- 11879451 TI - The UKCC's Scope of Professional Practice--some implications for health care delivery. AB - AIMS: The research reported here is part of a larger study commissioned by the United Kingdom Central Council for Nursing, Midwifery and Health Visiting (UKCC) to analyse and understand the impact of the Scope of Professional Practice (Scope) on the practice of nursing, midwifery and health visiting. The data in this paper relate to attitudes about, and perceptions of Scope among a variety of stakeholders. BACKGROUND: This research provided a valuable opportunity to explore the essence of Scope and the ways in which it can enable nurses, midwives and health visitors to adapt to growing or changing health care needs. The findings contribute to this debate, by drawing on the knowledge and experience of the key groups involved in the change process. METHOD: The study utilized several approaches, tailored to meet the requirements of each phase. In this (the final) phase a structured questionnaire was sent to different groups of stakeholders. FINDINGS AND CONCLUSIONS: The stakeholders displayed considerable interest in the development of innovative systems for delivering nursing and midwifery care, provided that the necessary safeguards and support were in place. Scope was seen as a valuable way of optimizing the skills and contribution of nurses, midwives and health visitors. PMID- 11879452 TI - Occupational stress in nursing: a review of the literature. AB - The management and reduction of occupational stress are recognized as key factors in promoting employee well-being. Nursing is one of the many disciplines contributing to a huge body of research into the causes and effect of the ill defined phenomenon of occupational stress. This literature review considers the ontological contribution of a number of disciplines to a growing body of knowledge on the subject of stress. The paper examines the complex issue of stress management, highlighting the impact of organizational culture and transformational leadership style on staff satisfaction levels. PMID- 11879453 TI - Knowledge of diabetes among personnel in home-based care: how does it relate to medical mishaps? AB - OBJECTIVE: To assess the influence of knowledge about diabetes on the performance of diabetes care for the elderly involving insulin treatment, with special attention to aspects of patient safety in home care. DESIGN: A questionnaire was administered to nurse's aides and assistant nurses (n = 3144). Answers to questions about knowledge of diabetes were related to "relevant" or "risky measures" as judged from a hypothetical diabetes case. A 94% response rate was obtained. The study took place in January 1997 in 15 of Sweden's 289 municipalities. RESULTS: Insufficient theoretical knowledge about how the blood sugar is related to an insulin reaction led to an almost threefold increased risk of taking a "risky measure". Insufficient knowledge about reasons for an insulin reaction also resulted in a higher risk, as was the case for personnel working in home based care in contrast to those working solely in Institutional care. In addition, the risk that a nurse's aide would take a "risky measure" was higher than that for an assistant nurse. This may indicate that the basic theoretical knowledge of nurse's aides is inadequate. CONCLUSION: Deficiencies in basic knowledge of diabetes among nurse's aides and assistant nurses constitute a major cause of potentially serious mishaps in home care of elderly diabetic patients treated with insulin. PMID- 11879458 TI - Developing scholarship in nursing in Britain -- towards a strategy. PMID- 11879459 TI - Values and evaluation in health care. AB - The purpose of this paper is to broaden the view of evaluation in health care by 'problemizing' the concepts of quality and evaluation and relating them to a more general discussion of values. The discussion of the concept of quality shows that the concept of quality is often vague or contradictory and that the relationship between quality and costs is problematic. The discussion is broadened by studying quality and evaluation from the viewpoint of four categories of values: scientific values, aesthetic values, ethical values and economic values. The authors also show that values, in addition to constituting the basis for evaluation, actually guide the whole process of care. Values are explicit and implicit elements of the care culture and the individual's action system. The authors conclude that the four value categories could be used to study which values actually guide the care process in real situations. PMID- 11879460 TI - Depending on the intent and emphasis of the supervisor, clinical supervision can be a different experience. AB - The paper proposes that clinical supervision can be a very different experience for the practitioner depending on the intent and emphasis of the supervisor. Utilizing Habermas's typology of knowledge constituted interests (1971), clinical supervision can be viewed as a dialectic between technical and emancipatory interests. The rhetoric and spirit of supervision would suggest an emancipatory approach yet the reality is that when supervision is accommodated with bureaucratic cultures, the technical interest will be dominant, especially when supervisors are in line-management roles to practitioners. This raises issues around the nature and role of clinical leadership, and ways of bridging the tension between emancipatory and technical interests. PMID- 11879461 TI - Contemporary nurses' uniforms---history and traditions. AB - This paper explores the literature related to the history and traditions of contemporary nurses' uniforms. It also presents the views of nurses about uniforms following the abandonment of a prescribed uniform for nurses working in a Professorial Nursing Unit in a large Hospital in Victoria. The literature dealing with the question of uniforms focuses on professionalism, status and power, infection control, identity, modesty, symbolism and occupational health and safety. The nurses who participated in the study expressed many of these themes. PMID- 11879462 TI - Community nurses', home carers' and patients' perceptions of factors affecting venous leg ulcer recurrence and management of services. AB - AIM: This study examines the feasibility of utilizing social service home carers (SSHC) to provide a collaborative approach with community nurses for the provision of leg ulcer aftercare in four National Health Service (NHS) Trusts. The purpose of this study was to gain insight into what community nurses and people with healed venous leg ulcers felt influenced leg ulcer recurrence. BACKGROUND: Studies have demonstrated that provision of community-based leg ulcer clinics has improved healing rates of venous leg ulcers, yet recurrence remains a problem. The reasons for this are far from clear, and further research is required before unequivocal support can be given to one approach to the provision of care for this client group. Collaborative approaches to the provision of leg ulcer aftercare are beginning to receive more attention. It has been recognized that social service health carers could be co-opted to provide essential aftercare once healing has occurred, although the logistics of this approach have not been fully explored. METHODS: This study was conducted in four NHS Trusts. Stage one used focus groups to explore the perceptions of district nurses (n = 15) and social service health carers (n = 15) of a leg ulcer shared care project and to gain insight into factors that they felt influenced recurrence. The second stage used semistructured interviews (n = 12) to explore the perceptions that people with healed leg ulcers have about factors influencing ulcer recurrence. FINDINGS: Key themes emerging from this study were: health promotion is perceived by community nurses and patients to be ineffective and leg ulcer aftercare services are fragmented. Organizational factors such as time constraints and limited resources were cited by community nurses and home carers as being responsible for high leg ulcer recurrence rates. Community nurses expressed a desire to delegate preventative aspects of leg ulcer care to home carers rather than participate in health promotion strategies to support healing behaviours. CONCLUSIONS: A strategy aimed at supporting healing behaviour in elderly people has the potential to reduce the recurrence of leg ulceration and improve quality of life. The findings suggest that such a strategy needs to rationalize delivery of leg ulcer aftercare to provide seamless care. It needs to improve carers' and patients' understanding of factors influencing leg ulcer recurrence and facilitate development of a more balanced professional--patient relationship. PMID- 11879464 TI - Changing government, changing cultures? Moving the quality agenda towards effective processes to counterbalance performance outcomes. AB - This paper begins by identifying current government initiatives to tabulate performance in the National Health Service by the identification and codification of selected clinical indicators, and notes the ambivalence as to whether effectiveness is to be seen on financial or qualitative terms. It continues by reviewing official and academic definitions of effectiveness, and discovers a similar lack of consensus. It then moves on to consider the concept of effectiveness in another public utility -- education. A distinction is drawn between effectiveness as a measure of institutional outcome and as a key factor influencing institutional process. The factors that promote effectiveness in education are identified and exemplified. The paper concludes by reflecting on the steps necessary if such processes are to be adopted in the management of the National Health Service, and notes that the new 'National Plan' offers, at least in theory, an ideal context for a collective effort to enhance effectiveness processes in the health service, in which managers can play a decisive role. PMID- 11879463 TI - Psychiatric nurses' conceptions of how a group supervision programme in nursing care influences their professional competence: a 4-year follow-up study. AB - AIM: The aim of the study was to describe, after 4 years, psychiatric nurses' conceptions of how a 2-year group supervision programme within nursing care had influenced their professional competence. BACKGROUND: The intention of group supervision in nursing care is to understand nurses' experiences within real care settings and to structure these in a professional and personal context. METHODS: Ten psychiatric nurses participated in a 2-year group supervision programme. They were interviewed 4 years after the group supervision was ended. Data were analysed according to the phenomenographic method. FINDINGS: Six description categories emerged: a feeling of job satisfaction; gaining knowledge and competence; gaining a sense of security in nursing situations; a feeling of personal development; realizing the value of supervision; and a sense of professional solidarity. CONCLUSIONS: The findings of the 4-year follow-up showed that a group supervision programme in nursing care had lasting influences on the psychiatric nurses' professional competence in the form of a pronounced professional identity and an integrated nursing care perspective. Group supervision contributes to maintaining the strength and energy needed to carry on working, which makes continuing supervision necessary. IMPLICATIONS: An important research implication could be to investigate the type of knowledge that ought to be developed within group supervision in nursing care. PMID- 11879466 TI - The role of the nurse manager in ensuring competence -- the use of portfolios and reflective writing. PMID- 11879467 TI - Values in the National Health Service: implications for nurse managers. AB - AIM: The values of an organization are key factors which influence the way it is managed. The purpose of this paper is to examine the values of the National Health Service and consider the implications they have for nurse managers. BACKGROUND: Three reports have been published recently which place values at the heart of the debate concerning the nature and purpose of the NHS. The development of these values is discussed and the clashes that arise between them are identified. CONCLUSION: It is argued that this situation presents an opportunity to strengthen the position of nurses in management and thus ensure health care management retains a patient focus. PMID- 11879468 TI - Cost--benefit analysis of team supervision: the development of an innovative model and its application as a case study in one Finnish university hospital. AB - AIM: To develop a model of costs and benefits of team supervision and a formula, which are examined more closely by means of an example. BACKGROUND: The popularity of clinical supervision (CS) as one of the methods of supporting health care practitioners' professional development (formative function), coping (restorative function) and quality improvement (normative function) has increased in the 1990s. CS may take the form of one-to-one or group supervision. Team supervision is a special form of group supervision. It means a group that has an interrelated work life outside the group. A host of literature and articles is available on CS. However, the costs and benefits of CS are less examined even though these have given rise to discussion particularly among decision-makers, because the monetary benefit of CS remains unsolved. METHOD: A nominal group technique was used to develop a model of costs and benefits of team supervision and a formula was derived on the basis of the model. The existing statistical data, for example a hospital ward's annual reports, data on sick days and reports on indemnities were utilized in the application of the formula. FINDINGS AND CONCLUSION: Team supervision was efficient in economic terms on the example ward. The model and the formula constitute a first attempt to ascertain the net present benefit of team supervision. Both the model and the formula need to be further tested, specified and refined. PMID- 11879469 TI - Leadership skills for the 21st century. AB - In today's ever-changing health care environment, nurses, especially administrators and managers, require leadership skills that provide direction for a new generation of nurses. Currently, there is a shortage of nurses globally. Along with this shortage of nurses, there is a shortage of nurse leaders. The importance of cultivating new nursing leaders is imperative for a healthful society. Gaining knowledge to become an effective nurse leader is one way to ensure quality health care for the future. PMID- 11879470 TI - The qualities of an empowered nurse and the factors involved. AB - BACKGROUND: In developing the quality of health care attention has recently focused on the professional progress of nurses. Simultaneously, empowerment ideology has been increasingly adopted as the conceptual framework for the development of nursing. The empowerment process has usually been described from an organizational starting point. In the present study empowerment is approached starting from the individual nurse's point of view. AIM: This study aims to explore (a) what an empowered nurse is like, (b) how he/she performs his/her functions, (c) what promotes empowerment, and (d) what prevents empowerment. METHOD: The material was collected by interviewing 30 nurses who had participated in a career advancement project at a university hospital in Finland. The data were analysed by qualitative content analysis. RESULTS: Five categories were discerned: moral principles, personal integrity, expertise, future orientation and sociability. These represented the performance and the qualities of an empowered nurse, as well as the promoting and preventing factors. CONCLUSIONS: Empowerment is a process steered by personal values and endeavours as well as by environmental factors. An empowered nurse possesses those qualities which make possible high self-esteem and successful professional performance and progress. PMID- 11879471 TI - Preventing the 'professional cleansing' of nurse educators. AB - The purpose of this paper is to argue that contrary to perceived wisdom nursing education ought to abandon the lecturer practitioner role on the grounds that it is a flawed concept predicated on a false assumption. In addition, and more seriously, it is in effect a significant hidden subsidy to service at the expense of education. It is suggested that by restructuring the nurse educator's role using the concepts of primary nursing as a method of organizing nurse educator's core activities, the perception and reality of teaching practice could be transformed. Calpin-Davies suggests that such a strategy enables nurse educators to be considered as practising nurses. It also has the added advantage that it is consistent with the form of organizing nursing expected of clinical colleagues. In addition it provides a basis for partnership with clinical nurses and with students, it responds to the imagined theory--practice gap, and affords nurse educators the means of becoming a credible role model. PMID- 11879473 TI - Evaluation of the QALY model for analysis of cost-effectiveness of eating training after stroke. AB - New methods developed and insights gained in research are of increasing significance in health care and the question is which services and methods are to be implemented. If eating training after stroke is to be implemented it must be given priority in relation to other measures. Otherwise there is a risk that patients with eating difficulties after stroke will be fed by personnel or permanently receive nourishment via tube. This may lead to expensive measures or costs for the tube feeding as well as costs for the personnel needed for feeding, and patients' well-being will be reduced if they do not have the opportunity to eat as independently as possible. Economic analyses should not guide the priorities to be made, but can be one of several bases for resource allocation. The estimation of cost-effectiveness must, however, be made by means of a method which in its ethical foundation is in line with nursing and the Swedish Government Bill for guiding priorities in health care. The aim of this paper is to discuss the ethical foundation of nursing care and the ethical principles proposed by the Swedish Government Bill for guiding priorities in health care and the model of Quality Adjusted Life Years (QALYs) related to eating training after stroke. The findings showed that there was a considerable difference in the ways health and health maximization were discussed in the QALY model and in specific nursing care. There are two aspects underpinning the QALY model that are not in line with either the ethical foundations of nursing or the principles proposed by the Swedish Government Bill for guiding priorities in health care. However, a new method called Equity Adjusted Life Years (EQALYs) can be a tool for evaluation in specific nursing care. Cost per EQALY is based on a compromise between initial severity of disease and treatment effect that is very close to the distribution rule applied in specific nursing care. In conclusion it is obvious that cost per EQALY, a balance between health maximization and severity of disease and treatment effects, can be a tool for the evaluation of eating training after stroke. PMID- 11879472 TI - Ward sisters' objectives in developing nursing and problems with development. AB - AIM: The study aimed at ascertaining the objectives, development strategies and problems of ward sisters in developing nursing. BACKGROUND: More knowledge is needed of first-line management in order to be able to develop organizations and ward sisters' professional training. METHODS: Data collection was by a questionnaire completed by 37 ward sisters in two Finnish hospitals. Thirty-six ward sisters responded to the open-ended questions and their answers were analysed using content analysis. FINDINGS: As for their objectives, ward sisters emphasized a functioning working community and good care. Good care was based on a variety of theoretical ideas, on the quality and economy of care. The means by which these objectives could be achieved were process thinking, reflection on action, leadership and the development of the working community. The leadership strategies mentioned were profit management and participatory leadership. In developing the working community, ward sisters relied on collaboration, good climate and training. Ward sisters' problems with developing nursing care were related to resources, work organization and the working community. CONCLUSIONS: Besides expertise in nursing, ward sisters are required to have skills in leading and developing the working community, collaboration and especially the ward's climate. PMID- 11879477 TI - Politics, power and control in the British National Health Service -- will anything really change? PMID- 11879478 TI - An appraisal of the use of secondment within a large teaching hospital. AB - INTRODUCTION: This study was undertaken in a large teaching hospital in Sheffield. It explores the use of secondment as a vehicle for practice, service and career development. AIM: To provide us with an understanding of the ways in which we utilize secondment opportunities, with a view to developing good practice guidelines that will help both the individual and the organization to maximize the potential in each secondment post. METHOD: A survey of nursing staff who had been on secondment during the previous year. Questionnaires were used to gather data from the senior nurse in each specialty directorate to develop an organizational (seconder) perspective and 20 secondees to provide an individual (secondee) perspective. RESULTS: Nurses tended to be seconded from clinical roles into specialist clinical roles or non-clinical roles, predominantly in areas of research, audit, practice development and teaching. Seconded posts were new roles for individuals and the majority were relocated to new work environments. Secondment was overwhelmingly seen as an opportunity, allowing individuals to develop new skills and knowledge, progress their career and gain a broader strategic perspective. However, there were a number of barriers to progress: lack of role definition for the organization and the individual; uncertainty about the future; falsely raised hopes that secondments would be extended; uncertainty about status; and difficulties adjusting to a new environment and culture within unrealistically short timeframes. CONCLUSIONS: Secondment use has become widespread throughout the National Health Service (NHS) and is a very positive and popular vehicle for staff and service development. The potential benefits are high but must be offset against the risks. This paper introduces an organizational risk assessment matrix which can be used to inform the development of effective secondment ventures. PMID- 11879479 TI - Emotional intelligence: a vital prerequisite for recruitment in nursing. AB - This paper explores Goleman's (1996) concept of 'emotional intelligence' in relation to recruitment to preregistration nurse education programmes. Current studies consistently demonstrate that emotional intelligence is the common factor which marks out individuals as leaders, innovators and effective managers. The role of the qualified nurse is evolving continually and 'portable' skills are the key qualities demanded by a health care system under pressure to compete. These include the ability to work effectively in teams, the ability to recognize and respond appropriately to one's own and others' feelings and the ability to motivate oneself and others. They are collectively termed 'emotional intelligence'. We believe we need selection processes that will determine levels of emotional intelligence in prospective candidates, as they could be a reliable predictor of success in both clinical nursing practice and academic study. Research indicates that emotional intelligence cannot be developed quickly enough through interpersonal skills training and therefore it is essential that nurse educators create assessment strategies that will identify emotional intelligence at recruitment. PMID- 11879480 TI - Peach, practice, placements and partnership: an initiative to support clinical placements in nursing curricula. AB - This article reports on the formation of an innovative new unit, termed the Clinical Placement Support Unit (CPSU), within a Faculty of Health in a Midlands university. The CPSU is concerned with managing the clinical experiences of the Faculty's health care students and promoting a healthy dialogue between the staff in the Faculty and their clinical colleagues. Several approaches by which a relatively small group of people (the Faculty staff) keep informed a relatively large group of people (the clinical staff within the local NHS Trusts) are detailed. One of the approaches is an assessors newsletter and a second is a website for use by both health care students and the clinical assessors of those students. The main part of the website is an ambitious attempt to help better prepare students prior to their clinical placements and this section of the website is called the Placement Directory. The Placement Directory currently holds details on hundreds of placements where the health care students may be placed. Each web page gives details about a specific clinical placement and has a number of hyperlinks to relevant internet resources. PMID- 11879481 TI - Focus group interviews to examine the role and development of the clinical nurse specialist. AB - This paper reports on the use of focus groups to investigate the development of clinical nurse specialist roles. The need for this project was identified during initial research, undertaken in two London Hospital Trusts, to assess the variety and nature of advanced clinical nursing roles already in evidence. From analysis of these data, this second stage of the research developed, in which the future of the clinical nurse specialist role within these two trust settings was explored. The initial steps of grounded theory, analytical techniques and procedures, using a constant comparative method, were used to analyse the findings from five focus groups (composed of a total of 25 nurses from two trusts). From the data, six categories were identified; role components; experience versus education; supportive strategies; personal qualities; future role development and development strategies. These categories provide the framework for discussing the findings of the research, within the context of the available literature. This examination of the clinical nurse specialist role highlights issues for career progression and education strategies. Both will need careful planning, in order for the clinical nurse specialist role to be considered and evolve as part of a potential strategy for the development of nursing roles within the two trusts and on a national level. PMID- 11879482 TI - An exploratory study in a community National Health Service Trust to understand why enrolled nurses choose not to convert to first-level registration. AB - AIM: The aim of this research study was to more fully understand at local level what it was that prevented enrolled nurses (ENs) coming forward for conversion to the first level of the United Kingdom Central Council for Nursing, Midwifery and Health Visiting (UKCC) nursing register. BACKGROUND: An NHS Trust had first-level nursing shortages and looked to the 69 ENs nurses -- 12% of the nursing workforce -- to meet that shortfall via conversion to first-level UKCC nurse registration. Based on local surveys, their intention to convert was the same as national findings, in that high numbers said they wanted to convert. In addition, this NHS Trust provided support and fully funded conversion courses, yet they did not come forward. METHODS: A qualitative approach based on focus groups with ENs was adopted to research this 'problem' within the local organizational context. FINDINGS: ENs were unaware that fully funded course places were available, extremely fearful of the academic expectations of the conversion course and highly committed to family needs. Conclusions Nationally, policymakers advocated the retention of and/or the conversion of ENs, on the grounds that research participants did not perceive or believe that managers were supportive of this proposal. KEY RECOMMENDATIONS: Create managers who looked beyond the immediate 'problem' of nursing shortages and invested in ENs to retain them in the NHS workforce. Establish local policy and an implementation plan to address the needs of ENs in line with clinical governance and the local nurse retention strategy. Empower ENs who would secure solutions to the issues for themselves. PMID- 11879483 TI - Brief encounters costing the NHS dear. PMID- 11879488 TI - How do expert mental health nurses make on-the-spot clinical decisions? A review of the literature. AB - An increasing number of writers have studied the work of Benner (1984) and Schon (1983) and have explored the concepts of expertise and 'intuitive' or 'tacit' knowledge. Efforts have been made to address the phenomenon of the theory- practice gap. Reborn interest in the craft of the professions is evident. A number of studies about the clinical decision-making process have suggested that clinicians are actually using a form of deductive reasoning known as hypothetico abductivism whilst reflecting in action. Exploration of expert decision-making by psychiatric nurses requires first that the nature of psychiatric expertise be clearly described. However, literature shows that this is not an easy task. In order to facilitate the development of expertise in novice practitioners, further research is required into the nature of expertise and expert decision-making. Cultivating and recording reflection in action around expert decision-making has revealed greater insights into the gap between theory and practice. It is evident that there is a need for a model that makes sense of the informal theory that is generated through practice. PMID- 11879489 TI - From iron gaze to nursing care: mental health nursing in the era of panopticism. AB - The purpose of this paper is to question the utilization of mechanical devices (cameras and microphones) to ensure the surveillance of hospitalized patients on psychiatric wards. The works of French philosopher, Michel Foucault, and those of nursing theorist, Jean Watson, are used to support this analysis. A growing number of Canadian psychiatric health care institutions are using mechanical devices for surveillance. The security of staff and patients as well as therapeutic purposes are stated as rationale for these practices. However, a Foucauldian perspective leads us to think otherwise. The metaphor of the panopticon is then used to uncover another reality: a disciplinary one. Within the scope of this paper, the question of surveillance, disciplinary power, caring philosophy, and mental health nursing will be examined. PMID- 11879490 TI - Imperative ideals and the strenuous reality: focusing on acute psychiatry. AB - The aim of this study was to describe the complexity of the working situation on an acute psychiatric ward as well as how nurses balance tensions between ideals and the reality of daily work. By means of field research, the study aimed to arrive at a deeper understanding of the reality that nursing staff and patients experience. The analysis shows that the acute and unpredictable character of the working situation in combination with short hospital stays results in a tentative and summary nursing care characterized by 'therapeutic superficiality'. This constitutes a hindrance to encountering the patient as a person. The demand on 'treatment effectiveness' creates work-related stress. Hence, a partly articulated conflict develops between the professional and humanistic ideals of psychiatric nursing and the strenuous reality that the staff have to adjust to. This conflict is solved in various ways, depending on whether they belong to the pragmatic, idealist, traditionalist or enforcer attitude in relation to the ward's mandate. The demand on treatment effectiveness seems to promote a medical model in the daily work, even though a humanistic and existential approach can be traced in the nurses' caring philosophy. PMID- 11879491 TI - Something special: forensic psychiatric nursing. AB - In this paper the claims that forensic psychiatric nursing has achieved the status of a specialist area of nursing are refuted. An examination of the literature demonstrates that specialist knowledge and skills have not been documented. It is suggested that three requirements are necessary if forensic psychiatric nurses wish to achieve specialty status. Forensic nurses have to consolidate their role in the containment and care of patients, they have to return to the nurse-patient relationship as the foundation of psychiatric nursing practice, then, within that relationship, nurses must expand their practice to include dealing with offence issues. PMID- 11879492 TI - A survey of community mental health nurses' perceptions of clinical supervision in Northern Ireland. AB - Since the early 1990s, clinical supervision has been the subject of debate by nurse academics and practitioners. This debate has encouraged the adoption of clinical supervision by the profession throughout the United Kingdom. Search of the literature demonstrates that there has been little published research regarding clinical supervision in Northern Ireland. This study is designed to redress this information deficit. The current position of clinical supervision in relation to community psychiatric nursing in Northern Ireland is explored and evaluated. A survey approach was adopted, collecting data from community psychiatric nurses (CPNs) in Health and Social Services Trusts in Northern Ireland. Data was obtained relating to the practice of clinical supervision and to attitudes of CPNs, their managers and supervisors. Results indicate that there is support for clinical supervision and that it is being implemented within community psychiatric nursing in Northern Ireland, although not in all cases. However, the findings indicate that serious education and training deficits exist, and the importance of the interface between managerial and clinical supervision is emphasized. The issues of providing effective education and training in supervision skills, and the uncertainty that was highlighted regarding fundamental concepts underpinning clinical supervision, have implications for nursing practice, education and management. In addition, based on findings of this study, the difference between management-led supervision and clinical supervision as envisaged by the UKCC, which promotes the personal and professional development of nurses, requires further exploration. PMID- 11879493 TI - Clients' reflections on relationships with nurses: comparisons from Canada and Scotland. AB - It has been suggested that the crucial elements in nursing situations are the nurse, the client, and what goes on between them. This paper examines what goes on between clients and nurses during interpersonal relationships, from the perspective of the clients. Data are presented from studies conducted in Canada and Scotland. It is shown that what clients want, or do not want, during relationships with their nurses, is similar on both sides of the Atlantic. The findings reported in this paper are relevant to transcultural nursing, ethical care, the growth in consumerism, and client advocacy. The findings suggest also that there is a need for nursing research to focus on clinical outcomes in order to establish whether clients' perceptions of helping relationships have any relevance to favourable health outcomes, and the evidence base for clinical nursing. PMID- 11879494 TI - The commonality and synchronicity of mental health nurses and palliative care nurses: closer than you think? Part one. AB - The role of the palliative care nurse emphasizes the need for holistic care, and as this role has developed it has become evident that palliative care nurses require skills which, arguably, not all registered general nurses possess; particularly, skills pertaining to the psychological, social and spiritual domains of the person. In order to identify the skills that such nurses may require, there may be merit in considering other specialities of nursing which pay particular attention to the psychological, social and spiritual domains of the person. Consequently, this two-part paper explores the areas of commonality and synchronicity between palliative care nurses and mental health nurses. The authors argue that this commonality is best articulated under the headings: defining the needs of the client group, the role of the nurse in non-physical care, the nurse--client relationship, and the locus of control. They also argue that the differences between these groups of nurses are best articulated under the headings: facilitation/confrontation, and the focus on physical care. Part one of this paper therefore focuses on the first three areas of alleged commonality, with part two focusing on the fourth commonality, the key differences and the implications of such similarity. Given these areas of similarity the authors argue there is a case for reconsidering if the RGN qualification is an essential requirement for working within palliative care or if those with other skills -- skills based on 'being with' rather than 'doing for' -- such as RMNs, should be thought of for such roles. PMID- 11879495 TI - The commonality and synchronicity of mental health nurses and palliative care nurses: closer than you think? Part two. AB - This is the second of a two-part paper which explores the areas of commonality and synchronicity between palliative care (PC) nurses and mental health nurses. The authors argue that this commonality is best articulated under the headings: defining the needs of the client group, the role of the nurse in non-physical care, the nurse--client relationship, and the locus of control. They also argue that the differences between these groups of nurses are best articulated under the headings: facilitation/confrontation and the focus on physical care. Part one focused on the first three areas of commonality, whereas this paper focuses on the fourth commonality, the locus of control. It also focuses on key differences and the implications of such similarity. The paper highlights the practice, education and research implications of this alleged commonality. It suggests, given the evidence that clients perceive the therapeutic relationship as the vital and unique aspect of PC nursing, that those working within palliative care need to question whether or not RGN registration is an essential requirement, or whether those with other skills, such as psychiatric/mental health (P/MH) nurses, should be considered for such roles. It highlights the need for the provision of post-basic counselling courses and the potential value for PC nurses of receiving clinical supervision from P/MH nurses or mental health liaison nurses. Lastly, it posits that the research issues arising out of this alleged commonality centre on the potential impact such transitions in care delivery may have on the care delivered, on the nurses themselves and on the clients. PMID- 11879496 TI - Depressed men: an exploratory study of close relationships. AB - Patients suffering from schizophrenia or bipolar affective disorder may progressively worsen and become severely disabled, and may then be classified as suffering from severe and enduring mental illness. Concern about risk to self and others focuses on this patient group, and community psychiatric nurses (CPNs) are under pressure to target patients with this diagnosis. CPNs have been accused of neglecting patients with a severe and enduring diagnosis in favour of other patient groups, but if they restrict services at primary care level this may have serious implications for patients. Patients who have had no previous contact with mental health services may have potentially serious and life threatening conditions. For example, depressed men may not be categorized as severely mentally ill, but the suicide rate amongst this patient group is very high, and they may externalize depression and resort to violence if untreated. Depressed men may lack social support and the means to express psychological distress, and these factors may precipitate or exacerbate depression. Early referral and assessment can prevent crises, deterioration in mental health and suicide. CPNs do not necessarily have to provide ongoing care following assessment, but they do have a significant role to play in primary health care referrals for the non psychotic mentally ill. CPNs act as a filter for the expertise and resources of multidisciplinary mental health teams. General practitioners will have increasing difficulty accessing the resources of multidisciplinary mental health teams if CPNs are unable to accept primary health care referrals. Primary care interventions are very important for the assessment of depression because they may help men to express psychological distress and assist them to access appropriate services and treatment. The experience of working with depressed men in a primary health care setting revealed that many lack confidants, or do not confide in those close to them. The interview schedule designed by Brown & Harris (1978) to gather data on the relationships of depressed women was used to explore the relationships of depressed men who were attending a Mental Health Day Centre. PMID- 11879497 TI - An exploratory investigation into the nursing management of aggression in acute psychiatric settings. AB - The management of aggression in inpatient psychiatric facilities is a major concern for nurses. Despite the prevalence of research on aggression within psychiatric settings, very few studies have tracked the clinical management of identified patients in a systematic manner. The overall aim of this study was to explore the clinical management of patients identified as potentially aggressive in psychiatric inpatient settings. A multimethod approach was used to collect the data. This included surveys, focus groups, and a file audit of patients identified from a random sample of completed aggression incident forms. The findings indicated that 88% of nurses who participated in the survey were assaulted and indicated the need for risk assessment; team management and review; and the importance of the first 5 days of admission as crucial for assessment and review. The development of support structures, formal preceptoring and supervisory arrangements and postincident supports were suggested as part of an overall strategy to support nurses working in acute inpatient settings. PMID- 11879498 TI - Have we the right to deny people their right to embrace their emotional pain? PMID- 11879499 TI - Qualitative and quantitative approaches in mental health nursing: moving the debate forward. A response to Rolfe, Parsons, Beech & Clarke. PMID- 11879506 TI - Tomorrow's Doctors: extending the role of public health medicine in medical education. PMID- 11879508 TI - Clinical skills training--practice makes perfect. PMID- 11879509 TI - Tomorrow's doctors: much quoted; but has it been fully implemented? PMID- 11879510 TI - Love it or hate it? Medical students' attitudes to computer-assisted learning. PMID- 11879511 TI - Reviewing intuitive decision-making and uncertainty: the implications for medical education. AB - CONTEXT: Intuition and uncertainty are inescapable conditions of many instances of clinical decision- making. Under such conditions biases and heuristics may operate, distorting the decision-making process. Physicians and students are generally unaware of these influences. PURPOSE: To review the extant literature regarding the role of uncertainty and intuition and associated biases on medical decision-making, to highlight the implications this holds for medical education. CONTENT: Using literature identified via Medline and Bioethicsline searches of the past 3 decades, this paper reviews the sources of uncertainty in clinical practice and the role of intuitive decision-making. A detailed description of associated heuristics and biases is provided, and linked with demonstrable examples from medical decision-making. CONCLUSIONS: It is argued that although uncertainty can be reduced, it can never be completely eliminated from decision making. Therefore most decision-making performed in medicine contains an irreducible intuitive element and is thus vulnerable to these biases and heuristics. Given that few medical curricula overtly address the process of medical decision-making, both medical students and physicians remain vulnerable to these effects on their own (and their patients') decision-making. Insight via education appears the major means in which to avoid distorting decision-making processes. PMID- 11879512 TI - Learning preferences, computer attitudes, and student evaluation of computerised instruction. AB - OBJECTIVE: To explore the relationship between learning preferences, attitudes towards computers, and student evaluation of a computer-assisted instructional (CAI) program. CONTEXT: A third year required clerkship in surgery at the University of Nebraska Medical Center. METHODS: A mixed-methods design combining attitudinal measures and qualitative interviews was employed to assess student reactions to a CAI program on angiography completed during a required surgical clerkship. Between January 1998 and July 1999, 151 students completed the program. Prior to participating, students completed the Rezler Learning Preference Inventory (LPI) and a computer attitudes survey (CAS). The LPI characterizes learning preferences as being abstract or concrete, individual or interpersonal, and student-structured or teacher-structured. The CAS measures attitudes towards computers and their role in education. After using the CAI program, students evaluated the module by completing an 18 item questionnaire. Based on LPI and CAS scores, 31 students were invited to participate in an in depth qualitative interview on their experiences and perceptions of the program. RESULTS: There was no relationship between learning preferences, computer attitudes, and evaluation of the CAI program. Students were very positive about the program's content, clarity, organization, and ease of use. They also rated it as efficient and effective. However, many still indicated a preference for lecture and text-based learning. Qualitative interviews suggest students worry computers will supplant student--teacher contact. CONCLUSIONS: Learning preferences and prior attitudes toward computers did not bias student reactions to the CAI program assessed in this study. However, students expressed concerns that CAI would interfere with the traditional student--teacher encounter and relationship. PMID- 11879513 TI - Early introduction of clinical skills teaching in a medical curriculum--factors affecting students' learning. AB - OBJECTIVE: To evaluate the effects of the early introduction of clinical skills teaching on students' learning following an overhaul of the curriculum of a traditional Asian medical school. METHODS: Randomly selected medical students in Year I and II were invited to participate in 30 focus group interviews while all students were asked to assist with the questionnaire survey. Most students were contacted personally to help them understand the objectives of the study. Confidentiality was emphasised and a non-faculty interviewer was recruited for the interviews. RESULTS: Two hundred and eight of Year I/Year II students attended the lunchtime focus group interviews (response rate=86.7%) while 252 (73.5%) students returned the questionnaire. The majority of them (87%) agreed or strongly agreed that it was good to introduce clinical skills in the early years of the curriculum. They reflected that the course enhanced their learning interest and made them feel like doctors. They also made many constructive suggestions on how the course could be improved during the interactive focus group interviews so that the negative effects could be minimised. CONCLUSION: It is useful to introduce clinical skills in the early years of a medical curriculum. A comprehensive course evaluation, using both quantitative and qualitative methods, helps to collect useful information on how the course can be improved. PMID- 11879514 TI - Does a rural educational experience influence students' likelihood of rural practice? Impact of student background and gender. AB - CONTEXT: The family medicine clerkship at the University of Calgary is a 4-week mandatory rotation in the final year of a 3-year programme. Students are given the opportunity to experience rural practice by training at 1 of several rural practices. OBJECTIVE: To determine whether exposure to a rural educational experience changes students' likelihood of doing a rural locum or rural practice and whether student background and gender are related to these practice plans. METHOD: Clinical clerks from the Classes of 1996-2000, who trained at rural sites, responded to questionnaire items both before and after the rural educational experience. Responses to the questionnaire items and discipline of postgraduate training served as dependent variables. Student background and gender were independent variables. RESULTS: As a result of the rural educational experience all students were more likely to do a rural locum. Compared to their urban-raised peers, students from rural backgrounds reported a significantly greater likelihood of doing a rural locum and practising in a rural community, irrespective of gender or participating in a rural educational experience. There was no relationship between background and career choice. CONCLUSION: A rural educational experience at the undergraduate level increases the stated likelihood of students participating in rural locums and helps to solidify existing rural affiliations. Students with rural backgrounds have a more favourable attitude toward rural practice. This pre-post study provides further support for the preferential admission to medical school of students with rural backgrounds to help alleviate the rural physician shortage. PMID- 11879515 TI - Evaluation of a family-oriented continuing medical education course for general practitioners. AB - OBJECTIVE: To explore the long-term effects of a 2-year Family Systems Medicine course. Fifteen experienced GPs participated in the training programme. SETTING: Continuing Education Centre, University of Tampere, Department of Public Health Science and General Practice, University of Oulu, Finland. METHODS: The participants assessed the development of their professional skills on the Doherty Baird scale and filled in 2 questionnaires. The material obtained from the application form and 2 questionnaires was analysed using the grounded theory method. RESULTS: The reasons for taking part in the course seemed to be the constant increase in the workload, problems caused by the demands for change and adaptation, stress and exhaustion. Furthermore, 10 health centres out of 15 had adopted the population-based practice, which requires different working methods compared to the old routines. Some trainees reported that their family-centred working methods improved during the course. A year after the end of the training, all of the GPs who had participated were using such methods in their daily practice. Half of the participants felt that they had also improved the functioning of their working group by making it more family-oriented. The significance of multi-professional collaboration was one of the most important insights during the course. CONCLUSION: The 2-year family-oriented training programme provided GPs with systemic thinking and with new skills, including the ability to work with families. The programme raised awareness of the need for multi-professional collaboration in the primary care settings. PMID- 11879516 TI - A performance assessment module for experienced general practitioners. AB - OBJECTIVE: To trial in Australian general practice a performance assessment module based on review of a sample of videotaped consultations. DESIGN: As part of a broader project 33 doctors provided 20 videotaped consultations collected according patient age, gender and problems encountered. The consultations were rated by a single trained rater. RESULTS: The consultations covered sufficient variety of patient ages, gender balance and problems encountered to achieve validity and the single rater design achieved defensible reliability (generalisability coefficient 0.86). CONCLUSION: The module is suitable for further exploration as an optional method for certification or re-certification assessment. PMID- 11879517 TI - Child health and obstetrics-gynaecology in a problem-based learning curriculum: accepting the limits of integration and the need for differentiation. AB - BACKGROUND AND OBJECTIVES: The University of Manchester's undergraduate curriculum was reorganised in 1994. Problem-based learning (PBL) was chosen as the central educational method throughout the 5 years. A thematic interdisciplinary style provided a framework around which to select and integrate content. The theme for family, reproductive and child health for the 14-week Families and Children Module (FCM) in year 4 integrated content from obstetrics gynaecology, paediatrics (including child psychiatry), genetics and public health. This paper focuses on the FCM which has put to test some of the principles of integration. METHODS: The educational process and outcome of the FCM was evaluated with information from (1) Open Forum feedback sessions (2) student questionnaires and (3) students' performance in an objective structured clinical examination (OSCE). RESULTS: Significant problems were encountered during the first year of the FCM (1997--98). An Open Forum for all students and staff was convened after each module in order to address the level of concern. Students' responses to questionnaires reflected the overall impression obtained from the Open Forum. Difficulties appeared to be more than the anxieties and challenges inherent in organisational change and were not simple practical timetabling problems. Tutors perceived a loss of coherence and student contact; students reported difficulties maintaining sufficient continuity of focus in clinical and academic learning experiences. The intended level of integration of paediatrics and obstetrics-gynaecology was unmanageable. Consequently, the FCM (1998-99) was divided into separate attachments for clinical learning and experience in paediatrics and obstetrics-gynaecology, each of 7 weeks' duration. Further open feedback sessions revealed that the modifications implemented in 1998-99 were associated with positive experiences for students and tutors. Comparison of responses to the evaluation questionnaires showed that students' overall rating of the module was higher (P=0.002) for 1998-99 (mean 3.4, SD 0.9) than for 1997-98 (mean 3.0, SD 1.1). CONCLUSIONS: We emphasise that horizontal integration can result in an experience of disintegration for students and tutors. Certain disciplines, such as paediatrics and obstetrics-gynaecology, may not have enough in common for full curriculum integration. PMID- 11879518 TI - Student perceptions of tutor skills in problem-based learning tutorials. AB - OBJECTIVE: The problem-based learning (PBL) tutor plays a role that is different from the role of a teacher in a conventional teaching format. In the Faculty of Medicine and Health Sciences, United Arab Emirates, all students are Arab nationals and tutors are expatriates with different sociocultural backgrounds from the students. This study was designed to investigate how students evaluate tutors in PBL tutorials and whether student evaluations of tutors change with the progress of students in PBL tutorials. METHODS: Differences in tutor performance evaluation by male and female students were also analysed. The students evaluated 12 tutor skills in a scale of 1-3, 1 being 'below average' and 3, 'outstanding'. Student responses from a total of 314 (98.1%) completed forms collected over 2 academic years were analysed statistically. A total of 14 tutors participated in the PBL programme. RESULTS: The analysis revealed that tutors as a group were rated as having average to outstanding tutor skills in 10 items of the evaluation form. Students and faculty perceptions were different for the tutor skills of guiding students for information management. The students expected more support from tutors, whereas the tutors tried to emphasize self-learning in the PBL curriculum. Lower scores to the tutors in the 'problem' bringing sociocultural and religious issues for discussion showed that a gap in sociocultural/religious understanding between students and tutors might influence tutor skills. CONCLUSIONS: Differences in tutor evaluation by male and female students indicate necessity of adopting different strategies by tutors in a different sociocultural background. The results of the study have direct implications for faculty development. PMID- 11879520 TI - The implementation of Tomorrow's Doctors. AB - BACKGROUND: In 1993 the General Medical Council published its recommendations on undergraduate medical education. AIM: To study the implementation of these recommendations in UK medical schools by means of informal visitations. METHODS: Teams consisting of 3-5 members visited the 25 UK medical schools in a 3-year period commencing in early 1995. RESULTS: Substantial changes have occurred in undergraduate medical education since the publication of Tomorrow's Doctors. Of the 13 principal recommendations, 3 had been implemented in most medical schools and a further 8 substantially implemented by the majority. However, progress in health promotion and the development of appropriate assessment schemes has been slower. CONCLUSIONS: Informal visits have served a useful purpose in monitoring the implementation of the General Medical Council's recommendations on undergraduate medical education. In addition, they have encouraged dialogue with the medical schools and allowed the identification of examples of good practice including the establishment by most schools of medical education units. PMID- 11879521 TI - The functional relationships of medical schools and health services. AB - Medical schools need good functional relationships with the health systems in which their students, graduates and staff will learn and work. With increasing complexity and rising costs of health delivery the 2 types of organisation have become more interdependent but not always as collaborating partners. Recent changes in the organisation of undergraduate medical education have been reactive to re-structuring of health services. For example, traditional teaching hospitals have become more specialised and less suitable for general medical education. While this has helped to decentralise medical education into appropriate clinical environments, these and other changes have generally been adaptive to the health systems rather than being instigated by the medical schools for educational reasons. This discussion paper examines the nature of the relationships and their contributory elements. A taxonomy of functional relationships is proposed with primary categories of (i) autonomy (ii) contiguity (iii) interdependence (iv) collaboration, and (v) obligation. It is suggested that medical schools should become more assertive in application of their creative talents and educational resources throughout the environment in which their graduates will use their medical skills, and in which they will work with other health care professionals. PMID- 11879519 TI - Is there a problem with the problems in problem-based learning? AB - OBJECTIVE: To determine the balance between acute and chronic medical problems in the PBL cases at 2 Australian medical schools. METHODS: Analysis of 162 PBL cases. RESULTS: Cases concentrate on acute problems in young people and neglect chronic disease in the old. CONCLUSION: This imbalance may contribute to negative attitudes towards elderly patients and people with chronic disease. PMID- 11879522 TI - Cardiopulmonary resuscitation training for undergraduate medical students: a five year study. AB - BACKGROUND: Cardiopulmonary resuscitation (CPR) training for undergraduate medical students and junior doctors has been noted to be poor in the past. Attempts have been made over the last decade to improve CPR training for all health professionals. AIM: This study aimed to determine if CPR training for undergraduate medical students in a single institution improved after initial concerns in 1992, and to observe trends in CPR training over five years. METHODS: Prospective single centre observational cohort survey by means of a 2-page self completed questionnaire to final year undergraduate medical students at the University of Glasgow (1993-97 inclusive). RESULTS: Mean annual response rate 58% (range 48%-67%). 99% of responders had been trained in basic life support during undergraduate training. The use of simulated arrests for training increased significantly. CPR training was concentrated in the first and final years. Training in all aspects of advanced life support increased, as did the students' confidence in these techniques. Student satisfaction with the amount of basic life support training increased very significantly and there was a small, but significant increase in student satisfaction with advanced life support training. Overall confidence at the prospect of being a member of the resuscitation team on qualification did not increase. CONCLUSIONS: There has been a sustained improvement in CPR training at this institution since 1993. Improvements in the training of specific advanced life support techniques does not lead to improved overall confidence in using these skills on qualification. Advanced life support training requires further expansion. PMID- 11879523 TI - Faculty opinions about a revised pre-clinical curriculum. AB - INTRODUCTION: Medical schools having innovative curricula have been encouraged to ascertain the levels of satisfaction of faculty members with the curriculum. Faculty at schools that employ problem-based learning (PBL) have been shown to have positive perceptions, but not all schools are in a position to adopt PBL on a large scale. This study sought to determine faculty members' opinions about a new curriculum that is less ambitious than one utilizing true PBL. CONTEXT AND SETTING: Since 1997, the University of Otago Medical School (Dunedin, New Zealand) has had an integrated, modular pre-clinical curriculum that emphasizes clinical relevance. It has proved popular with students. This study focused on faculty members' impressions. METHODS: We surveyed faculty members' opinions with a questionnaire identical to one used in studies at PBL schools. Faculty compared the students and their own levels of satisfaction in the old and new curricula on 7 to 10 items. The overall response rate was 85.4% (152 of 178). RESULTS: Perceptions of the new curriculum were positive among teachers who taught during the pre-clinical years and those who taught the students only after they reached the clinical years. Results for individual questions were in the same direction and generally similar in magnitude to those reported on identical items for PBL. CONCLUSION: We conclude that a hybrid curriculum that is more acceptable to many traditional teachers and students than is PBL has almost as great a positive effect on faculty members' perceptions of students' abilities and of the curriculum as does PBL. PMID- 11879526 TI - Re-annotation of genome microbial coding-sequences: finding new genes and inaccurately annotated genes. AB - BACKGROUND: Analysis of any newly sequenced bacterial genome starts with the identification of protein-coding genes. Despite the accumulation of multiple complete genome sequences, which provide useful comparisons with close relatives among other organisms during the annotation process, accurate gene prediction remains quite difficult. A major reason for this situation is that genes are tightly packed in prokaryotes, resulting in frequent overlap. Thus, detection of translation initiation sites and/or selection of the correct coding regions remain difficult unless appropriate biological knowledge (about the structure of a gene) is imbedded in the approach. RESULTS: We have developed a new program that automatically identifies biologically significant candidate genes in a bacterial genome. Twenty-six complete prokaryotic genomes were analyzed using this tool, and the accuracy of gene finding was assessed by comparison with existing annotations. This analysis revealed that, despite the enormous effort of genome program annotators, a small but not negligible number of genes annotated within the framework of sequencing projects are likely to be partially inaccurate or plainly wrong. Moreover, the analysis of several putative new genes shows that, as expected, many short genes have escaped annotation. In most cases, these new genes revealed frameshifts that could be either artifacts or genuine frameshifts. Some entirely unexpected new genes have also been identified. This allowed us to get a more complete picture of prokaryotic genomes. The results of this procedure are progressively integrated into the SWISS-PROT reference databank. CONCLUSIONS: The results described in the present study show that our procedure is very satisfactory in terms of gene finding accuracy. Except in few cases, discrepancies between our results and annotations provided by individual authors can be accounted for by the nature of each annotation process or by specific characteristics of some genomes. This stresses that close cooperation between scientists, regular update and curation of the findings in databases are clearly required to reduce the level of errors in genome annotation (and hence in reducing the unfortunate spreading of errors through centralized data libraries). PMID- 11879528 TI - Effect of simvastatin on bone markers in osteopenic women: a placebo-controlled, dose-ranging trial [ISRCTN85429598]. AB - BACKGROUND: Hydroxymethylglutaryl coenzyme A reductase inhibitors increase new bone formation in vitro and in rodents. Results of epidemiologic analyses evaluating the association between use of these cholesterol-lowering drugs, bone mineral density and fracture have been mixed. METHODS: Women (n = 24) with osteopenia, assessed by broad band ultrasound attenuation, were randomized to simvastatin 20 mg, 40 mg or identical-appearing placebo for 12 weeks. Fasting lipid profiles and biochemical markers of bone formation (bone-specific alkaline phosphatase) and resorption (N-telopeptides and C-terminal propeptide of type 1 collagen) were measured at baseline, 6 and 12 weeks. RESULTS: Plasma low density lipoprotein-cholesterol concentration fell 7%, 39% (p < 0.01 vs baseline) and 47% (p < 0.01 vs baseline) after 12 weeks of treatment with placebo, simvastatin 20 mg and 40 mg, respectively. At baseline, bone marker concentrations were similar in the three treatment groups. At 6 and 12 weeks, bone marker concentrations were not different from baseline, and no significant differences in bone marker concentrations were observed between treatment groups at either 6 or 12 weeks. CONCLUSION: Among osteopenic women, treatment with simvastatin for 12 weeks did not affect markers of bone formation or resorption. PMID- 11879527 TI - The use of breast conserving surgery: linking insurance claims with tumor registry data. AB - BACKGROUND: The purpose of this study was to use insurance claims and tumor registry data to examine determinants of breast conserving surgery (BCS) in women with early stage breast cancer. METHODS: Breast cancer cases registered in the Hawaii Tumor Registry (HTR) from 1995 to 1998 were linked with insurance claims from a local health plan. We identified 722 breast cancer cases with stage I and II disease. Surgical treatment patterns and comorbidities were identified using diagnostic and procedural codes in the claims data. The HTR database provided information on demographics and disease characteristics. We used logistic regression to assess determinants of BCS vs. mastectomy. RESULTS: The linked data set represented 32.8% of all early stage breast cancer cases recorded in the HTR during the study period. Due to the nature of the health plan, 79% of the cases were younger than 65 years. Women with early stage breast cancer living on Oahu were 70% more likely to receive BCS than women living on the outer islands. In the univariate analysis, older age at diagnosis, lower tumor stage, smaller tumor size, and well-differentiated tumor grade were related to receiving BCS. Ethnicity, comorbidity count, menopausal and marital status were not associated with treatment type. CONCLUSIONS: In addition to developing solutions that facilitate access to radiation facilities for breast cancer patients residing in remote locations, future qualitative research may help to elucidate how women and oncologists choose between BCS and mastectomy. PMID- 11879529 TI - The equivalence of numbers: the social value of avoiding health decline: an experimental Web-based study. AB - BACKGROUND: Health economic analysis aimed at informing policy makers and supporting resource allocation decisions has to evaluate not only improvements in health but also avoided decline. Little is known however, whether the "direction" in which changes in health are experienced is important for the public in prioritizing among patients. This experimental study investigates the social value people place on avoiding (further) health decline when directly compared to curative treatments in resource allocation decisions. METHODS: 127 individuals completed an interactive survey that was published in the World Wide Web. They were confronted with a standard gamble (SG) and three person trade-off tasks, either comparing improvements in health (PTO-Up), avoided decline (PTO-Down), or both, contrasting health changes of equal magnitude differing in the direction in which they are experienced (PTO-WAD). Finally, a direct priority ranking of various interventions was obtained. RESULTS: Participants strongly prioritized improving patients' health rather than avoiding decline. The mean substitution rate between health improvements and avoided decline (WAD) ranged between 0.47 and 0.64 dependent on the intervention. Weighting PTO values according to the direction in which changes in health are experienced improved their accuracy in predicting a direct prioritization ranking. Health state utilities obtained by the standard gamble method seem not to reflect social values in resource allocation contexts. CONCLUSION: Results suggest that the utility of being cured of a given health state might not be a good approximation for the societal value of avoiding this health state, especially in cases of competition between preventive and curative interventions. PMID- 11879530 TI - From immunoglobulin gene fingerprinting to motif-specific hybridization: advances in the analysis of B lymphoid clonality in rheumatic diseases. AB - In rheumatic diseases, autoantibody-producing cells of interest are often hidden in a polyclonal B-lymphocyte population. Immunoglobulin gene fingerprinting is a useful approach to screen for expanding clones and to detect recirculation between different locations. The gene fingerprinting approach and the Southern blot technique have been amalgamated, using electrophoretic transfer of a PCR product from an acrylamide gel onto a nylon membrane followed by hybridization with specific oligonucleotide probes. In contrast to conventional fingerprinting, the authenticity of immunoglobulin genes can be confirmed, individual genes can be detected and handling radionucleotides can be avoided. Also, the membrane may be reused for further investigations. PMID- 11879531 TI - Persistent Chlamydiae and chronic arthritis. AB - Urogenital infection with Chlamydia trachomatis can lead to development of an acute inflammatory arthritis, and this acute disease becomes chronic in some individuals. Research indicates that the organism is present in synovial tissue of patients with chronic disease in a persistent, rather than an actively growing, form. Importantly, metabolic and other characteristics of persistent Chlamydia differ from those of actively growing bacteria. Other studies suggest that Chlamydia pneumoniae can be found in a persistent state in the synovium and that it too may be involved in joint pathogenesis. These and other observations suggest a more complex role for the Chlamydiae in joint disease than previously recognized. This realization should engender a realignment of thinking among clinicians and researchers concerning both mechanisms of chlamydial pathogenesis in the synovium and design of new treatments for the disease. PMID- 11879532 TI - Ex vivo gene transfer in the years to come. AB - Synovial fibroblasts (SFs) have become a major target for ex vivo gene transfer in rheumatoid arthritis (RA), but efficient transduction of RA-SFs still is a major problem. The low proliferation rate and heterogeneity of RA-SFs, together with their lack of highly specific surface receptors, have hampered a more extensive application of this technique. Improving transduction protocols with conventional viral vectors, therefore, as well as developing novel strategies, such as alternative target cells, and novel delivery systems constitute a major challenge. Recent progress in this field will lead to the achievement of high transgene expression, and will facilitate the use of gene transfer in human trials. PMID- 11879533 TI - The Novartis-ILAR Rheumatology Prize 2001 Osteoarthritis: from molecule to man. AB - During our careers, we have developed new and innovative concepts pertaining to the pathophysiology of osteoarthritis which have assisted in the development of new therapeutic approaches. Moreover, our laboratory has long sought to develop protective agents for osteoarthritic structural joint tissues. The most significant concepts that have originated from our lab are briefly outlined in this commentary. PMID- 11879535 TI - Collagen of articular cartilage. AB - The extracellular framework and two-thirds of the dry mass of adult articular cartilage are polymeric collagen. Type II collagen is the principal molecular component in mammals, but collagens III, VI, IX, X, XI, XII and XIV all contribute to the mature matrix. In developing cartilage, the core fibrillar network is a cross-linked copolymer of collagens II, IX and XI. The functions of collagens IX and XI in this heteropolymer are not yet fully defined but, evidently, they are critically important since mutations in COLIX and COLXI genes result in chondrodysplasia phenotypes that feature precocious osteoarthritis. Collagens XII and XIV are thought also to be bound to fibril surfaces but not covalently attached. Collagen VI polymerizes into its own type of filamentous network that has multiple adhesion domains for cells and other matrix components. Collagen X is normally restricted to the thin layer of calcified cartilage that interfaces articular cartilage with bone. PMID- 11879536 TI - A critical appraisal of clinical practice guidelines for the treatment of lower limb osteoarthritis. AB - Clinical practice guidelines are important tools to assist clinical decision making. Recently, several guidelines addressing the management of osteoarthritis (OA) have been published. Clinicians treating patients with OA must ensure that these guidelines are developed with consistency and methodological rigour. We undertook a qualitative summary and critical appraisal of six medical treatment guidelines for the management of lower-limb OA published in the medical literature within the past 5 years. A review of these six guidelines revealed that each possesses strengths and weakness. While most described the scope and intended patient populations, the guidelines varied considerably in the rigour of their development, coverage of implementation issues, and disclosure of conflicts of interest. PMID- 11879534 TI - Lyme disease and current aspects of immunization. AB - Lyme disease is a tick-borne multisystem disease that affects primarily the skin, nervous system, heart and joints. At least three species of Borrelia burgdorferi sensu lato, namely Borrelia burgdorferi sensu stricto, Borrelia garinii, and Borrelia afzelii, can cause the disease. This review will focus mainly on the pathophysiology of Lyme arthritis, the long-term outcome of Lyme disease, and the recently licensed vaccine against Lyme disease. PMID- 11879537 TI - Th1 and Th2 cytokines regulate proteoglycan-specific autoantibody isotypes and arthritis. AB - BALB/c mice immunized with human cartilage proteoglycan (PG) develop arthritis accompanied by the production of autoantibodies to mouse cartilage PG. To determine whether the autoantibody isotype contributes to the onset and severity of arthritis, PG-specific serum IgG1 (Th2, IL-4-cytokine-supporting) and IgG2a (Th1, IFN-gamma-controlling) concentrations were monitored during immunization with PG in IL-4-deficient and IFN-gamma-deficient mice. Paradoxically, despite elevated IFN-gamma, the PG-specific IgG1 isotype was significantly higher than the PG-specific IgG2a response, and the PG-specific IgG1 isotype was independent of IL-4. In contrast, the serum concentration of PG-specific IgG2a isotype was six times higher in IL-4-deficient mice than in wild-type controls. Moreover, the high concentration of PG-specific IgG2a isotype in IL-4-deficient mice corresponded to an increased severity of arthritis. The concentration of PG specific IgG2a isotype was lower in IFN-gamma-deficient mice than in wild-type mice, and the incidence and severity of arthritis also were significantly lower. Concentrations of PG-specific IgG2a isotype autoantibody correlated with the onset and severity of arthritis, suggesting a pathological role of this isotype, probably locally in the joint. PMID- 11879538 TI - Volumetric computerized tomography as a measurement of periprosthetic acetabular osteolysis and its correlation with wear. AB - Osteolysis, which is considered to be a major source of morbidity following total hip joint replacement, has been notoriously difficult to measure accurately, particularly in the acetabular area. In order to study periacetabular osteolysis, specialized software for computerized tomography (CT) scan image analysis has been developed. This software (3D-CT) eliminates metal artifacts, allows three dimensional segmentation of the CT image, and reconstructs the segmented image to provide an accurate representation and measurement of volume for osteolytic lesions. In the present study, 20 patients underwent periacetabular osteolytic volume determination using 3D-CT, functional assessment (using the Harris Hip Scale, the Western Ontario and McMaster University Osteoarthritis Index, and the short form 36 questionnaire), and two-dimensional analysis of volumetic polyethylene wear using digitalized plain films. Periacetabular osteolysis correlated directly with the polyethylene wear rate (relative risk [RR] = 0.494, P = 0.027). If one patient with an acetabular revision, one patient with recurrent dislocation, and one patient with a Biomet prosthesis are excluded, then the correlation between wear and osteolysis is improved (RR = 0.685, P = 0.002). In summary, the current study demonstrates both the feasibility of CT imaging of periacetabular osteolysis and the correlation between polyethylene wear and osteolytic volume, providing a potential outcome measure for clinical trials that are designed to examine interventions in this complex disease process. PMID- 11879539 TI - Cytokine-stimulated T cells induce macrophage IL-10 production dependent on phosphatidylinositol 3-kinase and p70S6K: implications for rheumatoid arthritis. AB - IL-10 is an anti-inflammatory cytokine produced in the joint in rheumatoid arthritis by macrophages and infiltrating blood lymphocytes. Regulation of its expression is poorly understood, but previous findings have suggested that physical interactions with T cells may play a role. This report investigates signalling mechanisms involved in the production of macrophage IL-10 upon interaction with fixed, cytokine-stimulated T cells (Tck). Elutriated monocytes were differentiated to macrophages by macrophage-colony-stimulating factor (M CSF) and co-cultured with fixed T cells chronically stimulated in a cytokine cocktail of IL-2/IL-6/tumour necrosis factor (TNF)-alpha in the presence or absence of wortmannin and LY294002, inhibitors of phosphatidylinositol 3-kinase (PI3K), or of rapamycin, an inhibitor of p70 S6-kinase (p70S6K). Spontaneous IL 10 production by rheumatoid arthritis synovial-membrane mononuclear cells (RA SMCs) and co-cultures of rheumatoid arthritis T cells (RA-Ts) and macrophages was also assessed. RA-T and Tck induction of macrophage IL-10 production was suppressed by cell separation and inhibition of PI3K and p70S6K. PI3K involvement was also shown by phosphorylation of the downstream effector protein kinase B. Spontaneous IL-10 production by RA-SMCs was also inhibited by LY294002 and depletion of the nonadherent (T-cell-enriched) fraction of the cell population. IL-10 production in RA-SMCs and M-CSF-primed macrophages, activated by interaction with Tck, is PI3K- and p70S6K-dependent. PMID- 11879540 TI - Circulating cytotoxic CD8(+) CD28(-) T cells in ankylosing spondylitis. AB - Circulating CD8(+) CD28(-) T cells were found to be expanded more in patients with ankylosing spondylitis than in an age-matched healthy population (41.2 +/- 17.7% versus 18.6 +/- 7.6%). The level of CD8(+)CD28(-) T cells was dependent on the disease status, but was independent of age. Most of the CD8(+) CD28(-) T cells produced perforin after stimulation in vitro, in contrast to their CD8(+)CD28(+) counterparts. From the clinical perspective, the percentage of the cytotoxic CD8(+) CD28(-) T cells reflected a more severe course of disease, as it correlated with distinct movement restrictions, as well as the metrology score summarizing cervical rotation (in sitting position), chin-to-jugulum distance, thoracic Schober, chest expansion, and fingers-to-floor distance (P = 0.032). PMID- 11879541 TI - Staphylococcus aureus and Wegener's granulomatosis. AB - Wegener's granulomatosis (WG) is a form of systemic vasculitis. It is characterized by granulomatous inflammation in the upper and lower airways, vasculitis and necrotizing glomerulonephritis, and is strongly associated with antineutrophil cytoplasmic antibodies against proteinase 3. Since the etiology of the disease is not clear, treatment, consisting of corticosteroids and immunosuppressives, is nonspecific and associated with severe side effects. Pinpointing the trigger(s) of the disease would highly improve treatment. Clinical evidence shows that an infectious agent, the bacterium Staphylococcus aureus, is a risk factor for disease relapse, suggesting its involvement in the pathogenesis of WG. Here we review both clinical and experimental data that either indicate or support a role for S. aureus in WG. PMID- 11879542 TI - Clues to the etiology of autoimmune diseases through analysis of immunoglobulin genes. AB - The role of autoantibodies in the etiology of autoimmune diseases remains unclear. However, an examination of the sequences of these autoantibodies can be informative. Antibody sequences that violate constraints normally imposed during ontogeny and during development point to a failure of regulation. The existence of clonally related sequences indicates that production of these antibodies may frequently be driven by self-antigen. A better understanding of the mechanisms that normally constrain the composition of the antibody repertoire and of the nature of the inciting and/or driving antigens may yield new insights into both the pathogenesis and potential treatment of these crippling diseases. PMID- 11879543 TI - Commentary on "Genetic linkage and transmission disequilibrium of marker haplotypes at chromosome 1q41 in human systemic lupus erythematosus", by RR Graham et al. AB - Genome-wide linkage analysis studies in families with systemic lupus erythematosus (SLE) have revealed consistent evidence of linkage to several regions of the genome. In a previous issue of this journal, Graham and colleagues described their approach to following up the linkage data for one of these regions, 1q41-42. Using methods based on the transmission disequilibrium test, the region likely to harbour a SLE disease gene was refined to 2.3 Mb. This commentary discusses their approach and identifies lessons that may be applicable to the investigation of other complex diseases. PMID- 11879544 TI - Autoantibody systems in rheumatoid arthritis: specificity, sensitivity and diagnostic value. AB - The diagnosis of rheumatoid arthritis (RA) is primarily based on clinical symptoms, so it is often difficult to diagnose RA in very early stages of the disease. A disease-specific autoantibody that could be used as a serological marker would therefore be very useful. Most autoimmune diseases are characterized by a polyclonal B-cell response targeting multiple autoantigens. These immune responses are often not specific for a single disease. In this review, the most important autoantibody/autoantigen systems associated with RA are described and their utility as a diagnostic and prognostic tool, including their specificity, sensitivity and practical application, is discussed. We conclude that, at present, the antibody response directed to citrullinated antigens has the most valuable diagnostic and prognostic potential for RA. PMID- 11879546 TI - Body mass indices in patients with disabling hip osteoarthritis. AB - Hip osteoarthritis (OA) is a degenerative joint disease that results in substantial morbidity. The disease may be preventable in some instances by reducing risk factors associated with the disease. We undertook a study to determine whether being overweight or obese, a health risk that applies to younger and older age groups, is commonly associated with hip joint OA. The body mass indices (BMIs) of 1021 males and females ranging in age from 23 to 94 years and requiring surgery for end-stage hip joint OA were analyzed to find the prevalence of high body weights at the time of surgery. Being overweight was defined as having a BMI of 25-29.9 kg/m2 and being obese as having a BMI >30 kg/m2. BMIs indicative of overweight were recorded for 68% of the patients surveyed. Of 35 patients aged 30-39 years, 53.3% had BMIs >25, with a mean of 28.8, which nearly reaches the lower limit defined for obesity. On average, patients who had had previous surgery and complications warranting reimplantation of new surgical devices had BMIs in the obese range. Our findings suggest that a high percentage of patients with end-stage hip OA are overweight, including younger adults and those with symptoms of 3-6 months' duration. Moreover, patients whose BMIs are in the obese range may be at increased risk for removal and reimplantation of their prosthesis. PMID- 11879545 TI - The life cycle of chondrocytes in the developing skeleton. AB - Cartilage serves multiple functions in the developing embryo and in postnatal life. Genetic mutations affecting cartilage development are relatively common and lead to skeletal malformations, dysfunction or increased susceptibility to disease or injury. Characterization of these mutations and investigation of the molecular pathways in which these genes function have contributed to an understanding of the mechanisms regulating skeletal patterning, chondrogenesis, endochondral ossification and joint formation. Extracellular growth and differentiation factors including bone morphogenetic proteins, fibroblast growth factors, parathyroid hormone-related peptide, extracellular matrix components, and members of the hedgehog and Wnt families provide important signals for the regulation of cell proliferation, differentiation and apoptosis. Transduction of these signals within the developing mesenchymal cells and chondrocytes results in changes in gene expression mediated by transcription factors including Smads, Msx2, Sox9, signal transducer and activator of transcription (STAT), and core binding factor alpha 1. Further investigation of the interactions of these signaling pathways will contribute to an understanding of cartilage growth and development, and will allow for the development of strategies for the early detection, prevention and treatment of diseases and disorders affecting the skeleton. PMID- 11879547 TI - Cytokine mRNA and protein expression in primary-culture and repeated-passage synovial fibroblasts from patients with rheumatoid arthritis. AB - Constitutive mRNA expression and secretion of proinflammatory and anti inflammatory cytokines was comparatively analyzed in rheumatoid arthritis (RA) synovial fibroblasts (SFB), isolated from primary culture or derived by repeated passage; normal-skin fibroblasts were used as controls. First-passage RA-SFB (n = 3) secreted large amounts of IL-6 (15,800 +/- 2,110 pg/ml; mean +/- SEM), but only limited amounts of tumor necrosis factor (TNF)-alpha (22.1 +/- 1.1 pg/ml) or IL-10 (35.7 +/- 34.2 pg/ml; only one of three samples was positive). IL-1beta, IL 15, and IL-18 were not detectable at the protein level and showed very low mRNA levels by semiquantitative RT-PCR. In repeated-passage RA-SFB (tenth passage), protein secretion was significantly lower for IL-6 (one-twentieth of the initial level) and TNF-alpha (two-thirds), and markedly reduced for IL-10 (one-quarter, with only one of three samples positive). While the decrease of IL-10 protein from first to tenth passage was paralleled by a corresponding decrease of mRNA, the relative mRNA levels for IL-6 and TNF-alpha were actually increased (20-fold and 300-fold, respectively), indicating post-transcriptional and/or post translational regulation of these cytokines. Due to highly variable levels among individual patients, however, no significant differences were observed for any cytokine mRNA between primary-culture and repeated-passage RA-SFB (ninth passage). Likewise, no significant differences were detectable between RA-SFB and normal-skin fibroblasts (primary-culture and repeated-passage). By producing high amounts of IL-6 and limited amounts of TNF-alpha, RA-SFB may contribute to the (im)balance of proinflammatory and anti-inflammatory cytokines in the inflamed joint. PMID- 11879548 TI - Proinflammatory activity of TWEAK on human dermal fibroblasts and synoviocytes: blocking and enhancing effects of anti-TWEAK monoclonal antibodies. AB - Human tumour necrosis factor (TNF)-like weak inducer of apoptosis (hTWEAK) and two anti-hTWEAK mAbs were tested for their ability to elicit or block inflammatory responses in cultured human dermal fibroblasts and synoviocytes. Incubation with hTWEAK increased the production of prostaglandin E2, matrix metalloproteinase-1 (MMP-1), IL-6, and the chemokines IL-8, RANTES (regulated on activation, normal T expressed and secreted) and interferon-gamma-inducible protein-10 (IP-10) in culture supernatant of fibroblasts and synoviocytes. In combination with TNF or IL-1beta, hTWEAK further stimulated the secretion of prostaglandin E2, MMP-1, IL-6 and IL-8 up to fourfold, and IP-10 and RANTES up to 70-fold compared to TNF or IL-1beta alone. An anti-hTWEAK mAb, BCB10, blocked the effects of hTWEAK, whereas hTWEAK crosslinked by the anti-hTWEAK mAb, BEB3, further stimulated the inflammatory response of fibroblasts and synoviocytes. The anti-hTWEAK mAbs were ineffective in blocking or increasing the responses of TNF or IL-1beta and blocking anti-TNF mAb was ineffective in preventing the responses to TWEAK. These results were also confirmed at the RNA level for MMP-1, macrophage chemoattractant protein-1, RANTES, macrophage inflammatory protein 1alpha, IP-10 and IL-8. TWEAK in synergism with IL-1 and TNF may be an additional cytokine that plays a role in destructive chronic arthritic diseases. PMID- 11879549 TI - Autoantibodies directed to novel components of the PM/Scl complex, the human exosome. AB - The autoantigenic polymyositis/scleroderma (PM/Scl) complex was recently shown to be the human homologue of the yeast exosome, which is an RNA-processing complex. Our aim was to assess whether, in addition to targeting the known autoantigens PM/Scl-100 and PM/Scl-75, autoantibodies also target recently identified components of the PM/Scl complex. The prevalence of autoantibodies directed to six novel human exosome components (hRrp4p, hRrp40p, hRrp41p, hRrp42p, hRrp46p, hCsl4p) was determined in sera from patients with idiopathic inflammatory myopathy (n = 48), scleroderma (n = 11), or the PM/Scl overlap syndrome (n = 10). The sera were analyzed by enzyme-linked immunosorbent assays and western blotting using the affinity-purified recombinant proteins. Our results show that each human exosome component is recognized by autoantibodies. The hRrp4p and hRrp42p components were most frequently targeted. The presence of autoantibodies directed to the novel components of the human exosome was correlated with the presence of the anti-PM/Scl-100 autoantibody in the sera of patients with idiopathic inflammatory myopathy (IIM), as was previously found for the anti-PM/Scl-75 autoantibody. Other clear associations between autoantibody activities were not found. These results further support the conception that the autoimmune response may initially be directed to PM/Scl-100, whereas intermolecular epitope spreading may have caused the autoantibody response directed to the associated components. PMID- 11879550 TI - Expression of interleukin-18 receptor in fibroblast-like synoviocytes. AB - An excess of the proinflammatory substance IL-18 is present in joints of patients with rheumatoid arthritis (RA), and expression of IL-18 receptor (IL-18R) regulates IL-18 bioactivity in various cell types. We examined the expression of IL-18R alpha-chain and beta-chain and the biologic effects of IL-18 in fibroblast like synoviocytes (FLS) after long-term culture. The presence of both IL-18R chains was a prerequisite for IL-18 signal transduction in FLS. However, all FLS cultures studied were either resistant or barely responsive to IL-18 stimulation as regards cell proliferation, expression of adhesion molecules ICAM-1 and vascular cell adhesion molecule (VCAM)-1, and the release of interstitial collagenase and stromelysin, IL-6 and IL-8, prostaglandin E2, or nitric oxide. We conclude that the presence of macrophages or IL-18R+ T cells that can respond directly to IL-18 is essential for the proinflammatory effects of IL-18 in synovitis in RA. PMID- 11879551 TI - Mammographic breast density as a biomarker of effects of isoflavones on the female breast. AB - Isoflavones possess both estrogenic and anti-estrogenic actions, and are hypothesized to protect against breast cancer. However, two intervention studies of markers of proliferation on breast tissue have raised concerns that soy isoflavones may have an estrogenic effect on breast tissue. Increased mammographic breast density is associated with an elevated risk of breast cancer, although the mechanism underlying this relationship has not been explained. Estrogens increase and anti-estrogens decrease breast density. Breast density may therefore serve as a biomarker of estrogenic or anti-estrogenic effects of a treatment on breast tissue. The effect of isoflavones on breast density is under investigation. PMID- 11879552 TI - E-cadherin and loss of heterozygosity at chromosome 16 in breast carcinogenesis: different genetic pathways in ductal and lobular breast cancer? AB - Loss of heterozygosity at the long arm of chromosome 16 is one of the most frequent genetic events in breast cancer. In the search for tumour suppressor genes that are the target of loss of heterozygosity at 16q, the E-cadherin gene CDH1 was unveiled by the identification of truncating mutations in the retained copy. However, only lobular tumours showed E-cadherin mutations. Whereas investigations are still devoted to finding the target genes in the more frequent ductal breast cancers, other studies suspect the E-cadherin gene to also be the target in this tumour type. The present article discusses the plausibility of those two lines of thought. PMID- 11879553 TI - Distinct functions of BRCA1 and BRCA2 in double-strand break repair. AB - Individuals carrying BRCA mutations are predisposed to breast cancer. The BRCA1 and BRCA2 proteins are required for homologous recombination and DNA break repair, leading to the suggestion that they act in concert. However, direct evidence of a stable BRCA1/BRCA2 complex has not been demonstrated. Rather, the two proteins have been found as constituents of discrete, but perhaps nonexclusive complexes that are critical for repair. We discuss the interaction of BRCA1 with the BACH1 and BARD1 proteins, and suggest that the pleiotropic nature of mutations in BRCA1 may be associated with defects in protein--protein interactions. In contrast, the role of BRCA2 in DNA repair may be more defined by its direct interaction with the RAD51 recombinase. PMID- 11879554 TI - Cyclin D1 and mammary carcinoma: new insights from transgenic mouse models. AB - Cyclin D1 is one of the most commonly overexpressed oncogenes in breast cancer, with 45-50% of primary ductal carcinomas overexpressing this oncoprotein. Targeted deletion of the gene encoding cyclin D1 demonstrates an essential role in normal mammary gland development while transgenic studies provide evidence that cyclin D1 is a weak oncogene in mammary epithelium. In a recent exciting development, Yu et al. demonstrate that cyclin D1-deficient mice are resistant to mammary carcinomas induced by c-neu and v-Ha-ras, but not those induced by c-myc or Wnt-1. These findings define a pivotal role for cyclin D1 in a subset of mammary cancers in mice and imply a functional role for cyclin D1 overexpression in human breast cancer. PMID- 11879555 TI - Direct effects of bisphosphonates on breast cancer cells. AB - In addition to inhibiting bone resorption, bisphosphonates have also been shown to exhibit antitumour effects. In vitro, bisphosphonates inhibit proliferation and induce apoptosis in cultured human breast cancer cells. In addition, bisphosphonate treatment interferes with breast cancer cell adhesion to bone matrix, and inhibits cell migration and invasion. The combination of bisphosphonates with other anticancer drugs such as the taxoids markedly enhances these effects. These newly recognized direct actions of bisphosphonates on breast cancer cells indicate that these agents may have a greater role to play in treatment of patients suffering from cancers with a propensity to metastasize to bone. PMID- 11879556 TI - The present and future role of bisphosphonates in the management of patients with breast cancer. AB - At least 25% of patients with breast cancer develop skeletal metastases, with bone the site of disease producing the greatest morbidity. It is apparent that the bisphosphonates present an important component of the treatment strategy. They are now the treatment of choice in tumour-induced hypercalcaemia, and they can reduce bone pain and skeletal complications such as pathological fractures. In addition, bisphosphonates are being increasingly evaluated in the prevention of bone metastases and to prevent and treat cancer therapy-induced osteoporosis. Ongoing research is aimed at trying to define the optimum route, dose, schedule and type of bisphosphonate. PMID- 11879557 TI - Development of bisphosphonates. AB - Bisphosphonates are synthetic compounds characterized by a P[bond]C[bond]P group, and are thus analogs of inorganic pyrophosphate. They are used in medicine mainly to inhibit bone resorption in diseases like osteoporosis, Paget's disease and tumor bone disease. They have been used for over a century in industry, and only in 1968 was it shown that bisphosphonates have biological effects. These effects consist mainly of an inhibition of bone resorption and, when given in large amounts, an inhibition of ectopic and normal calcification. While the latter effect is the consequence of a physical-chemical inhibition of calcium phosphate crystal formation, the former is due to a cellular effect involving both apoptosis of the osteoclasts and a destruction of the osteoclastic cytoskeleton, inducing a decrease in osteoclast activity. The biochemical basis of these effects for the nitrogen-containing compounds is an inhibition of the mevalonate pathway caused by the inhibition of farnesylpyrophosphate synthase, which leads to a decrease of the formation of isoprenoid lipids such as farnesylpyrophosphate and geranylgeranylpyrophosphate. The other bisphosphonates are incorporated into the phosphate chain of ATP-containing compounds so that they become non hydrolyzable. The new P[bond]C[bond]P-containing ATP analogs inhibit cell function and may lead to apoptosis and death of osteoclasts. PMID- 11879558 TI - Actions of bisphosphonates in animal models of breast cancer. AB - The skeleton is the most common site of breast cancer metastases. These bone metastases are usually osteolytic and cause significant morbidity. Bisphosphonates, potent inhibitors of bone resorption, reduce skeletal morbidity in breast cancer patients with bone metastases. Animal studies with bisphosphonates are crucial to understanding the mechanisms by which these compounds affect bone and tumor cells in vivo. Such animal models of breast cancer that are used to test the efficacy of bisphosphonates are discussed. These studies may offer insight into the treatment of other tumor types that frequently metastasize to bone. PMID- 11879559 TI - Comparison of various characteristics of women who do and do not attend for breast cancer screening. AB - BACKGROUND: Information regarding the characteristics and health of women who do and do not attend for breast cancer screening is limited and representative data are difficult to obtain. METHODS: Information on age, deprivation and prescriptions for various medications was obtained for all women at two UK general practices who were invited to breast cancer screening through the National Health Service Breast Screening Programme. The characteristics of women who attended and did not attend screening were compared. RESULTS: Of the 1064 women invited to screening from the two practices, 882 (83%) attended screening. Screening attenders were of a similar age to non-attenders but came from significantly less deprived areas (30% of attenders versus 50% of non-attenders came from the most deprived areas, P < 0.0001) and were more likely to have a current prescription for hormone replacement therapy (32% versus 19%, P < 0.0001). No significant differences in recent prescriptions of medication for hypertension, heart disease, hypercholesterolaemia, diabetes mellitus, asthma, thyroid disease or depression/anxiety were observed between attenders and non attenders. CONCLUSION: Women who attend the National Health Service Breast Screening Programme come from less deprived areas and are more likely to have a current prescription for hormone replacement therapy than non-attenders, but do not differ in terms of age or recent prescriptions for various other medications. PMID- 11879560 TI - BRCA1 and BRCA2 mutations in a population-based study of male breast cancer. AB - BACKGROUND: The contribution of BRCA1 and BRCA2 to the incidence of male breast cancer (MBC) in the United Kingdom is not known, and the importance of these genes in the increased risk of female breast cancer associated with a family history of breast cancer in a male first-degree relative is unclear. METHODS: We have carried out a population-based study of 94 MBC cases collected in the UK. We screened genomic DNA for mutations in BRCA1 and BRCA2 and used family history data from these cases to calculate the risk of breast cancer to female relatives of MBC cases. We also estimated the contribution of BRCA1 and BRCA2 to this risk. RESULTS: Nineteen cases (20%) reported a first-degree relative with breast cancer, of whom seven also had an affected second-degree relative. The breast cancer risk in female first-degree relatives was 2.4 times (95% confidence interval [CI] = 1.4-4.0) the risk in the general population. No BRCA1 mutation carriers were identified and five cases were found to carry a mutation in BRCA2. Allowing for a mutation detection sensitivity frequency of 70%, the carrier frequency for BRCA2 mutations was 8% (95% CI = 3-19). All the mutation carriers had a family history of breast, ovarian, prostate or pancreatic cancer. However, BRCA2 accounted for only 15% of the excess familial risk of breast cancer in female first-degree relatives. CONCLUSION: These data suggest that other genes that confer an increased risk for both female and male breast cancer have yet to be found. PMID- 11879561 TI - Mammary cancer and epithelial stem cells: a problem or a solution? AB - The existing paradigms for stem cells in adult tissues include the integument, the alimentary canal, the lung, the liver, skeletal muscle and bone marrow. The mammary gland, by contrast, is the 'new kid on the block'. What little is known about stem cells in the mammary gland indicates that they possess a prodigious capacity for self-renewal. More importantly, in rodents, they persist with undiminished reproductive vigor throughout the organism's lifetime without regard to age or reproductive history. Do these stem cells represent primary targets for mammary neoplasia? If so, what are the implications for prevention/therapy? PMID- 11879562 TI - The utility of ductal lavage in breast cancer detection and risk assessment. AB - Ductal lavage (DL) permits noninvasive retrieval of epithelial cells from the breast. Clinical development of this technique has been fueled largely by its potential, as yet unproven, to improve detection of breast cancer and definition of individual risk for development of breast cancer. Early studies demonstrate the feasibility of performing this technique, provide data on cellular yield and findings, and demonstrate the ability to measure molecular markers in DL fluid. However, the sensitivity and specificity of DL for the detection of breast cancer remains unknown, as does the significance of atypia, particularly mild atypia, when found in DL fluid. Although DL appears safe and the device is approved by the US Food and Drug Administration, DL is still best utilized in the setting of clinical trials designed to resolve issues of sensitivity, specificity, and localization. PMID- 11879563 TI - Are Trp53 rescue of Brca1 embryonic lethality and Trp53/Brca1 breast cancer association related? AB - Brca1 is involved in multiple biological pathways including DNA damage repair, transcriptional regulation, and cell-cycle progression. A complex pattern of interactions of Brca1 with Trp53 has also emerged. Xu and coworkers found that haploid loss of Trp53 significantly reduces the embryonic lethality observed in mice with a homozygous in-frame deletion of Brca1 exon 11. They report that widespread apoptosis correlates with the embryonic lethality resulting from this homozygous delta11 Brca1 mutation. A mechanism responsible for Brca1-associated carcinogenesis is proposed. These experiments extend our knowledge of a complex Brca1/Trp53 relationship. However, the precise mechanisms through which Brca1 interacts with Trp53 to suppress mammary tumor formation have yet to be elucidated. PMID- 11879564 TI - The contribution of the environment (especially diet) to breast cancer risk. AB - Environmental factors play an important role in breast carcinogenesis. Opportunities for prevention are limited, however, because most of the known or suspected risk factors are not targets for modification. Dietary factors have generally not emerged as crucial contributors to mammary tumor causation. We still appear to be missing a critical piece of the breast cancer puzzle because we can only explain a moderate proportion of international and national variation in breast cancer rates. Research needs to pursue new avenues, focusing on exposure windows that have not yet been sufficiently explored, such as events between conception and adolescence, and on modifiable risk factors that show large variation within or between populations. PMID- 11879565 TI - How does the estrogen receptor work? AB - In breast cancer, interruption of estrogen receptor (ER)-alpha function is an effective therapeutic strategy. Despite the clinical benefit of interruption of ER-alpha function, the precise biological action of ER-alpha in breast tumors is not completely understood. Results of a recent study show that ER-alpha promotes growth of breast cancer cells by targeting expression of signaling components of the insulin-like growth factor system. Intriguingly, the authors of this study raise the possibility that unliganded ER-alpha itself may affect gene expression and breast cancer biology, and they suggest a potential mechanism for ER-alpha to stimulate proliferation in breast cancer. PMID- 11879566 TI - The role of cytokines in regulating estrogen synthesis: implications for the etiology of breast cancer. AB - Cytokines, such as IL-6 and tumor necrosis factor (TNF)-alpha, have an important role in regulating estrogen synthesis in peripheral tissues, including normal and malignant breast tissues. The activities of the aromatase, estradiol 17beta hydroxysteroid dehydrogenase and estrone sulfatase are all increased by IL-6 and TNF-alpha. Prostaglandin E2 may also be an important regulator of aromatase activity in breast tumors. Macrophages and lymphocytes, which invade many breast tumors, are thought to be an important source of factors that can stimulate estrogen synthesis in malignant breast tissues. The co-ordinated stimulation of the activities of the enzymes that are involved in estrogen synthesis offers an explanation for the high concentrations of estrogens that are present in breast tumors. PMID- 11879568 TI - Learning to remember: generation and maintenance of T-cell memory. AB - Immunologic memory results from a carefully coordinated interplay between cells of the immune system. In this review, we explore various aspects of the nature, generation, and maintenance of T lymphocyte-mediated immunologic memory. In light of the demonstrated heterogeneity of the memory T-cell pool, we hypothesize that subsets of memory T cells instructed to mature to distinct differentiation stages may differ, not only in functional and homing properties, but also in the conditions they require for survival, including antigen persistence and cytokine environment. Hence, according to this hypothesis, distinct memory T-cell subsets result from the nature and timing of the signals provided by the immune environment and occupy distinct niches. Intracellular and extracellular molecular mechanisms that underlie and modulate T-cell memory are discussed. PMID- 11879567 TI - The p53 pathway in breast cancer. AB - p53 mutation remains the most common genetic change identified in human neoplasia. In breast cancer, p53 mutation is associated with more aggressive disease and worse overall survival. The frequency of mutation in p53 is, however, lower in breast cancer than in other solid tumours. Changes, both genetic and epigenetic, have been identified in regulators of p53 activity and in some downstream transcriptional targets of p53 in breast cancers that express wild type p53. Molecular pathological analysis of the structure and expression of constituents of the p53 pathway is likely to have value in diagnosis, in prognostic assessment and, ultimately, in treatment of breast cancer. PMID- 11879569 TI - HIV-1, NFAT, and cyclosporin: immunosuppression for the immunosuppressed? AB - The human immunodeficiency virus-1 (HIV-1), the cause of AIDS, remains a significant cause of morbidity and mortality throughout the planet. Although reverse transcriptase and protease inhibitors have substantially slowed the virus, viral resistance complicates therapy. Because HIV-1 relies on its host's transcriptional machinery for its own replication, strategies for targeting activation-dependent transcription factors in CD4 T cells are being considered for adjunctive therapy in HIV-1-infected individuals. The nuclear factor of activated T cells (NFAT) family of transcription factors is one such target. On T cell stimulation, NFAT proteins translocate to the nucleus, where they activate a large number of early response genes, including cytokines such as interleukin-2. Activation and nuclear translocation of NFAT proteins are abrogated by the powerful immunosuppressants cyclosporin A (CsA) and FK506. Over the last several years, various investigators have demonstrated that NFAT proteins bind to the HIV 1 LTR promoter and increase viral transcription. In this report, further evidence supporting a role for NFAT proteins in augmenting HIV-1 transcription is presented. In addition, other mechanisms of HIV-1 inhibition by CsA are reviewed, and the rationale for the use of CsA to treat AIDS is discussed. PMID- 11879570 TI - Induction of EGFR-dependent and EGFR-independent signaling pathways by ultraviolet A irradiation. AB - Most of the signal pathways involved in ultraviolet (UV)-induced skin carcinogenesis are thought to originate at plasma membrane receptors. However, UVA-induced signal transduction to downstream ribosomal protein S6 kinases, p70(S6K) and p90(RSK), is not well understood. In this report, we show that UVA stimulation of the epidermal growth factor receptor (EGFR) may lead to activation of p70(S6K)/p90(RSK) through phosphatidyl isositol (PI)-3 kinase and extracellular receptor-activated kinases (ERKs). Evidence is provided that phosphorylation and activation of p70(S6K)/p90(RSK) induced by UVA were prevented in Egfr(-/-) cells and were also markedly inhibited by the EGFR-specific tyrosine kinase inhibitors AG1478 and PD153035. Furthermore, EGFR tyrosine kinase inhibitors and EGFR deficiency significantly suppressed activation of PI-3 kinase and ERKs in regulating activation of p90(RSK)/p70(S6K) but had no effect on activation of c-Jun NH(2)-terminal kinases (JNKs) and p38 kinase in response to UVA. Thus, our results suggest that UVA-induced EGFR signaling may be required for activation of p90(RSK)/p70(S6K), PI-3 kinase, and ERKs but not JNKs or p38 kinase. PMID- 11879571 TI - In vivo roles of RORalpha and Sp4 in the regulation of murine prosaposin gene. AB - Prosaposin has a central role in intracellular glycosphingolipid catabolism and also has extracellular functions. This locus is regulated temporally and spatially. The highest mRNA expression occurs in the central nervous system (CNS) and reproductive system. In vitro, the CNS-expressed proteins Sp4 and RORalpha bind to Sp1 and RORE sites within a 310-bp fragment directly upstream of the transcription start site. These transcription factors exhibit negative cooperativity in vitro for prosaposin expression. Mice deficient in RORalpha and Sp4 (Staggerer [Sg(-/-)] and Sp4 knockout [Sp4 KO], respectively) containing selected prosaposin promoter deletion transgenes were used in comparative expression studies to evaluate this negative cooperativity in vivo. Constructs containing the RORE or Sp1/U cluster alone were independently stimulatory. Deletion of the Sp1/U site led to a decrease in reporter activity only in the cerebellum of Sg(-/-) mice. The deletion of RORE and Sp1/U sites did alter the increase of reporter activity in the brain and eye, but not in the spinal cord, of Sg(-/-) mice. These results indicate that Sp4 and RORalpha play minor and major roles, respectively, in regional expression of the prosaposin locus in the brain, whereas expression in the spinal cord is independent of RORalpha. PMID- 11879572 TI - Hepatopoietin acts as an autocrine growth factor in hepatoma cells. AB - Hepatopoietin (HPO) is a novel human hepatotrophic factor. Its known function is mainly limited to supporting liver regeneration. Recently, it was shown by our laboratory that HPO acts as a mitogen for hepatoma cell lines and that there are HPO-specific receptors on the surface of these cells (Wang, G., et al., J Biol Chem 1999;274:11469-11472), indicating that HPO might be involved in oncogenesis in the liver. To study this hypothesis, we first conducted experiments in vitro to identify the existence of an autocrine loop of HPO/HPO receptor in hepatoma cell lines. It was demonstrated that HPO was actually expressed by hepatoma cells, such as HepG2, Bel 7402, and SMMC-7721, and secreted into the culture medium. Furthermore, it was shown that HPO-neutralizing antibody has an inhibitory effect on the uptake of tritiated thymidine by hepatoma cells. The results strongly suggest that HPO acts as an autocrine factor for hepatoma cells in vitro. PMID- 11879573 TI - Intravirion display of a peptide corresponding to the dimer structure of protease attenuates HIV-1 replication. AB - Current treatment of HIV-1-infected individuals involves the administration of several drugs, all of which target either the reverse transcriptase or the protease activity of the virus. Unfortunately, the benefits of such treatments are compromised by the emergence of viruses exhibiting resistance to the drugs. This situation warrants new approaches for interfering with virus replication. Considering the activation of protease in the virus particles, a novel strategy to inhibit HIV-1 replication was tested targeting the dimerization domain of the protease. To test this idea, we have selected four residues from the C terminus of HIV-1 protease that map to the dimer interface region of the enzyme. We have exploited Vpr to display the peptides in the virus particles. The chimeric Vpr exhibited expression and virion incorporation similar to wildtype Vpr. The virus derived from the HIV-1 proviral DNA containing chimeric Vpr sequences registered a reduced level of replication in CEM and CEM X 174 cells in comparison with viruses containing wildtype Vpr. Similar results were observed in a single-round replication assay. These results suggest that the intravirion display of peptides targeting viral proteins is a powerful approach for developing antiviral agents and for dissecting the dynamic interactions between structural proteins during virus assembly and disassembly. PMID- 11879574 TI - Model system for evaluation of alternative splicing: exon skipping. AB - Alternative splicing of the primary RNA transcript is a common mechanism for generating protein diversity. A model system was developed to study this process in vitro that is useful for evaluation of splicing of transcripts expressed in cells that do not grow well in culture. The system was used to analyze skipping of exon 4 of the amelogenin message, normally expressed in ameloblast cells for a short interval during tooth enamel development. Amelogenins are highly conserved proteins resulting from extensive alternative splicing, with domains involved in a range of functions, including mineral formation and intercellular signaling. In the bovine gene, the very short intron 4 was predicted to inhibit inclusion of exon 4, because in murine ameloblasts, exon 4 is detectably included in mRNA, and intron 4 is longer than the bovine counterpart. Bovine intron 4 was lengthened, and this size increase enhanced exon 4 inclusion sixfold to eightfold, although splice site selection was inaccurate. Intron length, therefore, is not the sole determinant controlling amelogenin exon 4 inclusion, and cis-acting inhibitory elements may also be involved in exon skipping. This vector system allows evaluation of splicing of a tissue-specific RNA by focusing on exons of interest through transfection of heterologous cultured cells without complications attributable to background transcription of the gene being evaluated. PMID- 11879575 TI - Cloning and functional analysis of pyruvate kinase promoter region from Drosophila melanogaster. AB - Pyruvate kinase (PK; EC 2.7.1.40) is a key glycolytic enzyme of Drosophila melanogaster. It catalyzes the conversion of phosphoenolpyruvate into pyruvate with the transfer of a phosphate group to ADP to form ATP. The ATP provides energy for cell growth and metabolism, and pyruvate participates in many metabolic reactions. Therefore, PK plays an important role in cell metabolism. Southern blot analysis, PCR, and sequencing were used to determine the content of a Drosophila pyruvate kinase (Pyk) genomic clone, lambdaPK61. The results indicated that the insert of lambdaPK61 comprised 8330 bp upstream of and 7186 bp downstream of the transcription start point of the Pyk gene. The size of the insert was 15,516 bp in total, which contained six genes including Pyk. Deletion mapping was applied to identify the promoter region and cis-acting elements 5' of PyK. Ten serial deletions produced by PCR were inserted upstream of the reporter gene (LacZ) to form recombinant plasmids, which were then transfected into Drosophila S2 cells. The results revealed that the regions -1475 approximately 1033 and -1033 approximately -534 of the 5' end of PyK possessed positive regulatory function for Pyk expression; i.e., increased gene expression. There were redundant putative cis-acting elements, including ecdysone response element (EcRE), E74A, and broad complex zinc finger (BRCZ) binding sites. Both E74A and BRCZ belong to the early genes regulated by ecdysone. This result suggested that Pyk might be regulated by ecdysone, directly or indirectly. However, the results of the developmental profile of Pyk expression by Northern blot analysis suggested that the effects of ecdysone on Pyk were repressive, not inductive. In addition, it was found that in these regions, there were many cis-acting elements related to egg and embryo development. Both -258 approximately -254 and -167 approximately -163 contained a CAAT box, and deletion of these regions decreased reporter gene expression. Therefore, it is suggested that both CAAT boxes are functional and that the promoter of Pyk might be located in the region of -258 approximately +109. No TATA box or downstream promoter element were identified around the transcription start site of Pyk. Additionally, PyK might share a regulatory region with an unknown neighboring gene. It was concluded that Pyk has the characteristics of a housekeeping gene. PMID- 11879576 TI - Decreased expression of retinol-binding proteins is associated with malignant transformation of the ovarian surface epithelium. AB - We have developed a modified form of suppression subtractive hybridization (SSH) that allows multiple specimens of distinct phenotypic groups to be compared for consistent differences in gene expression. We applied this system to identify genes that were expressed in normal rat ovarian surface epithelial (ROSE) cells but whose expression was lost/downregulated in four independently transformed rat ovarian cancer cell lines. Northern blot analysis using 14 of 28 nonredundant cDNA fragments from this difference library showed that the mRNA transcripts were present in normal ROSE cells but lost or markedly downregulated in four related transformed cell lines. Of particular interest, cellular retinol-binding protein 1 (CRBP1) and retinol-binding protein (RBP), two genes whose products are involved in retinol transport and metabolism, were found to be downregulated in this ovarian cancer model system. To determine if this change had relevance to human ovarian cancer, we evaluated a series of human ovarian cancer cell lines and a limited number of frozen human ovarian tumors and found lost or decreased expression of CRBP1 and RBP relative to expression in human ovarian surface epithelial (HOSE) cells. We hypothesize that the loss of CRBP1 and RBP expression disrupts retinol metabolism and retinoic acid production, which may facilitate the occurrence of genetic damage leading to the malignant transformation of the ovarian surface epithelium, the cells from which ovarian cancer arises. PMID- 11879577 TI - Morphologic change and elevation of cortisol secretion in cultured human normal adrenocortical cells caused by mutant p21K-ras protein. AB - In our previous study on the tumorigenesis of human functional adrenal tumors, we observed a high frequency of K-ras point mutations in clinical specimens. Furthermore, we cloned the mutated K-ras gene from the tumors and inserted it into vectors to transfect normal bovine adrenocortical cells to express the mutated K-ras gene. The mRNA level of steroidogenic enzymes such as cholesterol sidechain cleavage enzyme (P450SCC), 17alpha-hydroxylase/17,20-lyase (P450c17), and 3beta-hydroxysteroid dehydrogenase (3betaHSD) in the mutant K-ras stably transfected cells were elevated. Cultured normal adrenocortical cells from donors and patients with adrenocortical tumors were then transfected with mutant K-ras expression plasmids constructed from human adrenal tumors. Stable transfectants grew faster than normal cells. Additionally, morphologic change was observed in the transfected cells. Moreover, when the synthesis of hormones was analyzed, the mRNA of P450SCC, P450C17, and 3betaHSD was found to have increased, and the level of cortisol was 18 to 25 times that in control cells. The increased steroid hormone production in mutant K-ras-transfected cells was reversed by lovastatin, a pharmacologic inhibitor of p21ras function. These results, combined with previous reports of steroidogenic K-ras in bovine adrenocortical cells, suggest that the K-ras oncogene is involved in steroidogenesis in human adrenocortical cells. PMID- 11879578 TI - Promoter-binding activity of inflammation-responsive transcription factor SAF is regulated by cyclic AMP signaling pathway. AB - The serum amyloid A activating factor (SAF) was identified as a family of inducible transcription factors that is activated by many mediators of inflammation. Its activation involves a phosphorylation event, whose mechanism is not fully understood. Here, we show that cAMP treatment of several cell types, including mouse liver-derived BNL CL.2, human monocyte-derived THP-1, and a primary culture of vascular smooth muscle cells from porcine aorta, activated cellular SAF's ability to bind DNA. The protein kinase A (PKA) activity in cytoplasmic extracts of cAMP-treated cells was responsible for the potentiation of the DNA-binding activity of the cellular SAF proteins. Furthermore, treatment of nuclear extracts of untreated cells with purified PKA increased the DNA binding activity of cellular SAF proteins, and specific inhibitors of PKA abrogated the enhanced DNA-binding ability of SAF in the cAMP-treated cells. Consistent with these findings, overexpression of the catalytic subunit of PKA markedly increased expression of the SAF-regulated promoter. These results imply a functional role for the previously detected protein-protein interaction between SAF-1 transcription factor and the catalytic subunit of PKA and further demonstrate the consequences of cAMP-mediated signaling for the expression of SAF regulated genes. PMID- 11879579 TI - Mutational analysis of the phosphorylation sites of the Aie1 (Aurora-C) kinase in vitro. AB - We previously reported two novel serine/threonine kinases, Aie1 (mouse) and AIE2 (human), both later referred to as aurora-C, a newly recognized member of the mitotic aurora kinase family. In the present study, we analyzed the phosphorylation sites of mouse Aie1 by site-directed mutagenesis. Our results showed that protein kinase A (PKA) phosphorylates Aie1 at a threonine residue located at amino acid position 171. The T171A and T175A mutants, in which threonines located at residues 171 and 175 were replaced by alanines, revealed a significant increase in their kinase activities to phosphorylate ACS-1 (Aurora-C substrate 1). In contrast, the double mutant T171A-T175A showed impaired kinase activity. In addition, we had previously identified a PEST-like motif located at the N terminus of Aie1. Mutation analysis in the present study revealed that the quadruple mutant in which the PEST-like motif was mutated significantly abrogated Aie1 kinase activity. This is the first report of the analysis of potential phosphorylation sites of mouse aurora-C in vitro. PMID- 11879580 TI - Expression of type 2 cystatin genes CST1-CST5 in adult human tissues and the developing submandibular gland. AB - Type 2 cystatins comprise a class of cysteine peptidase inhibitor presumed to mediate protective functions at various locations, including the oral cavity. Seven cystatin genes are clustered within a 300-kb region of human 20p11.2. "Salivary" cystatins, encoded by CST1, 2, 4, and 5, are present in saliva at significant levels but have also been reported in other secretions, such as tears, suggesting that during their evolution, these genes have acquired mechanisms directing differential tissue-specific expression. However, their patterns of expression, which might also provide additional clues to their individual functions, have not been determined. Gene-specific RNase protection assays were used to examine the qualitative and quantitative distribution of expression of these seven genes within a collection of 23 adult human tissues. The CST3 gene, encoding cystatin C, was expressed at modest levels in all tissues examined. The presumptive pseudogenes CSTP1 and CSTP2 were not expressed at detectable levels in any tissue. The CST1, 2, 4, and 5 genes were expressed in differential, tissue-specific patterns. Expression of CST2 and CST5 was restricted to the submandibular and parotid glands, while CST1 and CST4 were expressed in these tissues and in the lacrimal gland. Immunohistochemistry studies localized expression to the serous-type secretory end pieces. Coexpression of CST1 and CST4 was also observed in the epithelial lining of the gallbladder and seminal vesicle. The CST1 product was detected in the tracheal glands and CST4 in the kidney and prostate. Despite their different adult patterns of expression, analysis of CST1, 2, 4, and 5 mRNA levels in infant submandibular glands demonstrated a coordinate upregulation of expression of between 3.5 and 9 months of age. The patterns of cystatin gene expression are consistent with several proposed oral functions of the salivary cystatins but also suggest they are important in other locations and that, despite their close sequence similarity, they are individually specialized. PMID- 11879582 TI - Acquired ciliary circumscribed grey hair (ACCG). AB - Grey-haired areas usually occur due to aging or inheritance. A case is described of abrupt occurrence of a focal circumscribed grey-hair in the eyebrow region (a single hair) in a 27-year-old woman. The phenomenon was named acquired ciliary circumscribed grey-hair (ACCG). Qualitative and semiquantitative findings were obtained by microanalytical studies. In addition to morphological differences from control hair, the ACCG hair showed a high percentage of sulfur (99.8%) and absence of oligoelements. PMID- 11879581 TI - Iron induces proliferation and morphogenesis in primmorphs from the marine sponge Suberites domuncula. AB - Dissociated cells from marine demosponges retain their proliferation capacity if they are allowed to form special aggregates, the primmorphs. On the basis of incorporation studies and septin gene expression, we show that Fe3+ ions are required for the proliferation of cells in primmorphs from Suberites domuncula. In parallel, Fe3+ induced the expression of ferritin and strongly stimulated the synthesis of spicules. This result is supported by the finding that the enzymatic activity of silicatein, converting organosilicon to silicic acid, depends on Fe3+. Moreover, the expression of a scavenger receptor molecule, possibly involved in the morphology of spicules, depends on the presence of Fe3+. We conclude that iron is an essential factor in proliferative and morphogenetic processes in primmorphs. PMID- 11879583 TI - Prevalence of bacterial vaginosis in adolescent girls. AB - The age-related increase in the prevalence of bacterial vaginosis was determined in adolescent virginal and sexually active girls aged 13 to 18 years. Higher rates were recorded for girls with two or more sexual partners. Considering the potential spectrum of consequences of bacterial vaginosis, preventive strategies are required in this age group, not only for bacterial vaginosis but also for other sexually transmitted diseases. PMID- 11879584 TI - Tinea incognito caused by trichophyton mentagrophytes -- a case report. AB - A case of widespread tinea due to Trichophyton mentagrophytes is described. A healthy 75-year-old woman presented with 134 typical tinea corporis and faciei lesions previously treated with topical steroids. The diagnosis was based on direct mycologic examination and culture. The treatment with oral administration of terbinafine for four weeks and topical application of clotrimazole resulted in complete clinical resolution of the lesions PMID- 11879585 TI - Eosinophilic fasciitis associated with simple traumatism. AB - Eosinophilic fasciiitis is an idiopathic and uncommon condition, a scleroderma like disorder that predominantly affects the extremities and is characterized by marked thickening and inflammation of the fascia, associated with with peripheral blood eosinophilia, hypergammaglobulinemia, and characteristic histologic findings. A case is presented of a 75-year-old female patient who sought medical assistance for sudden occurrence of induration and erythema of both lower extremities one week after she had fallen from a one-meter height, with generalization of the signs. The diagnosis of eosinophilic fasciitis was made on the basis of histopathologic findings, and treatment with glucocorticoids, non steroid anti-inflammatory drugs and antihistaminics was prescribed. PMID- 11879586 TI - Prostaglandins in dermatology. AB - Prostaglandins, local hormones, are produced by almost every tissue and organ in the body (except for erythrocytes). They are active at the site of their production or next to the cells that secrete them. Prostaglandins play an important biological role, because they are involved in many physiological and pathological processes. Some of their effects are still unknown. They are involved in inflammatory process of the skin. They have been implicated as possible mediators of contact allergic dermatitis and among others, act in immune response. The use of prostaglandin in dermatology is still experimental. In the near future, they might be use in therapy for systemic scleroderma, aberration of extremity circulation with ulcers, vasculitis, etc. An overview is provided of prostaglandins in dermatology. PMID- 11879587 TI - Andrej Procopchuk (1896 - 1970) and the development of dermatology and venerology in Byelorussia. AB - A short review is given of the development of dermatology and venerology in Byelorussia. The scientific activity of Professor Andrej Procopchuk (1896 - 1970), who was the founder of the scientific school of dermatology and venerology in Byelorussia, was of prime importance in this respect PMID- 11879588 TI - Cholangiocarcinoma of the Hepatic Hilum (Klatskin Tumor). AB - Since the description by Klatskin in 1965, the management of patients with adenocarcinoma of the hepatic bile duct bifurcation is viewed as a challenging clinical problem with a relatively poor prognosis. Surgery continues to be the mainstay of therapy. Complete resection of the tumor with negative histologic margins offers the best possibility of long-term survival, and hepatic resection is a critical component of the operative approach. Adjuvant chemoradiotherapy has failed to provide a significant survival benefit. Orthotopic liver transplantation for otherwise unresectable lesions remains controversial, as tumor recurrence has been reported in more than 90% of patients. With the shortage of organs, such patients to be selected carefully for transplanation. For patients who present with widespread disease and those with high operative risks, advances in interventional radiology and endoscopy have facilitated nonsurgical management options. Biliary decompression using expandable metallic stents provides superior patency and decreased frequency of hospitalization when compared with plastic stents. Moreover, patients treated with expandable metal stents have survival rates comparable with those who undergo surgical decompression, with fewer early complications. The benefit of external beam radiotherapy for palliation of proximal cholangiocarcinoma is uncertain. Radiotherapy in conjunction with biliary stenting has a survival benefit over stenting alone, but is not without potential toxicity. It should be considered as an adjunct to biliary decompression in all patients with good performance status, because modern conformal CT-based dosimetry can minimize toxicity to normal adjacent tissue. Photodynamic therapy is emerging as a new palliative treatment modality for patients with unresectable tumors in whom stenting has failed. It offers the advantage of an endoscopic delivery system, and unlike radiotherapy, photodynamic therapy may be delivered repeatedly. PMID- 11879589 TI - Mirizzi Syndrome. AB - The complete and definitive treatment of patients with Mirizzi syndrome is surgical. The treatment goals are the removal of the gallbladder with the offending stone(s) and the repair of the bile duct defect. A high index of suspicion for early recognition of this condition is paramount to prevent bile duct injury. Biliary anatomy is delineated precisely by preoperative and intraoperative imaging tests. The dissection of the gallbladder is conducted in an antegrade, fundus-first fashion. Extensive dissection of Calot's triangle is avoided. Instead, the gallbladder is opened, the impacted stone(s) is removed backward, and the confirmation of the presence a cholecystocholedochal fistula is established by direct inspection. A coexistent gallbladder carcinoma is excluded by taking frozen sections. In patients without biliary fistula (Mirizzi type I), simple cholecystectomy suffices to relieve the bile duct obstruction. In patients with biliary fistula (Mirizzi type II), the size of the fistula determines the type of repair. In general, small fistulas are repaired by choledochoplasty using a cuff of gallbladder remnant, whereas large bile duct defects require bilioenteric reconstruction (Roux-en-Y hepaticojejunostomy or choledochoduodenostomy). In patients unfit for surgery, biliary decompression is effectively accomplished by placement of stents using endoscopic or percutaneous techniques. Lithotripsy and removal of the offending stone can also be carried out in patients with Mirizzi type II. In general, nonsurgical treatment of Mirizzi syndrome is incomplete and places the patients on a path of intensive follow-up, multiple procedures, and the risk to continue suffering from complications of symptomatic gallstone disease. However, nonsurgical treatment allows for valuable time to prepare high-risk patients for a more elective and safer operation. PMID- 11879590 TI - Biliary Complications After Liver Transplantation. AB - The incidence of biliary complications after liver transplant is estimated to be 8% to 20%. Post-liver transplant biliary complications may lead to acute and chronic liver injury. The early recognition and prompt treatment of such complications improves the long-term survival of the patient and graft. An understanding of the type of biliary reconstruction, the rationale for creating a particular anastomosis, and the technical difficulties in reconstructing the biliary tract are important in assessing and managing complications after liver transplant. Because the clinical presentation of these patients may be subtle, the physician must be aggressive and thoughtful in ordering and interpreting the diagnostic tests. Important points to remember are 1) that noninvasive examinations may fail to detect small obstructions or leaks, 2) a liver biopsy often is performed prior to cholangiography to exclude rejection and ischemia, and 3) the liver biopsy can miss an extrahepatic obstruction by misinterpreting portal inflammation as rejection. Biliary leaks and strictures are the most common biliary complications following liver transplant. Less common complications include ampullary dysfunction and stone/sludge formation. The effective management of biliary complications following a liver transplant depends on understanding the natural history, the prognosis, and the available therapeutic options for each type of complication. PMID- 11879591 TI - Postoperative Bile Duct Strictures. AB - The management of patients with postoperative biliary stricture is a challenging problem that spans across several disciplines. A team approach involving endoscopists, interventional radiologists, and biliary surgeons is crucial for the successful management of individuals with this complex problem. Hepaticojejunostomy is the therapy of choice. Prolonged stenting, either through the percutaneous route or through the endoscopic retrograde route, is an alternative. Stricture recurrence is not infrequent and requires lifelong follow up. PMID- 11879592 TI - Uninvestigated Dyspepsia. AB - Dyspepsia, which is defined as pain or discomfort centered in the upper abdomen, is encountered frequently in primary care and subspecialty practice. Dyspepsia is a symptom complex caused by a heterogeneous group of disorders and diseases. A large fraction of patients with dyspepsia suffer from functional dyspepsia, in which no evidence of organic disease (typically on the basis of upper endoscopy) is found to explain persistent or recurrent symptoms. Initial management strategies for uninvestigated dyspepsia include empiric antisecretory therapy, the "test-and-treat" strategy for Helicobacter pylori, or prompt upper endoscopy. The cost-effectiveness of empiric therapy versus the test-and-treat strategy is dependent upon a number of variables including the prevalence of H. pylori infection, ulcer prevalence, and likelihood that an ulcer is due to H. pylori infection. As the prevalence of H. pylori infection falls and the likelihood of H. pylori negative ulcer increases, empiric antisecretory therapy will become more cost-effective. Upper endoscopy should be reserved for patients older than 45 to 50 years with symptom presentation and those with warning signs. Endoscopy also should be considered in those for whom empiric therapy or an attempt at the test-and-treat strategy fails. Common-sense dietary counseling can be helpful in patients with meal-related symptoms. Highly restrictive diets rarely improve symptoms and may be counterproductive if nutrition is compromised. PMID- 11879593 TI - Rescue Therapy for Helicobacter pylori. AB - Up to 35% of patients infected with Helicobacter pylori fail to respond to standard anti-H. pylori therapy. With the rising prevalence antimicrobial resistance, the failure rates of conventional proton pump inhibitor-containing triple therapy are expected to increase. Pretreatment antibiotic resistance testing should be done whenever possible to allow for tailoring of the treatment regimens. The data on second-line or rescue therapy are limited and usually are subjected to various biases and confounding factors. Switching between clarithromycin and metronidazole should be considered if repeated courses of proton pump inhibitor-containing triple therapy are used as second-line therapy in the absence of antimicrobial sensitivity testing. The prolongation of therapy duration with proton pump inhibitor, amoxicillin, and clarithromycin is ineffective for clarithromycin-resistant strains. The bismuth-containing quadruple therapy is the best salvage treatment in the absence of pretreatment antibiotic susceptibility. Furazolidone quadruple therapy (where available) and rifabutin triple therapy are salvage therapies of last resort. If these regimens fail, culture and susceptibility testing is required. PMID- 11879594 TI - Dumping Syndrome. AB - The dumping syndrome consists of early postprandial abdominal and vasomotor symptoms, resulting from osmotic fluid shifts and release of vasoactive neurotransmitters, and late symptoms secondary to reactive hypoglycemia. Effective relief of symptoms of dumping syndrome can be achieved with dietary modifications to minimize ingestion of simple carbohydrates and to exclude fluid intake during ingestion of the solid portion of the meal. More severely affected individuals may respond to agents such as pectin and guar, which increase the viscosity of intraluminal contents, or to drugs such as the alpha-glucosidase inhibitor acarbose, which blunts the rapid absorption of glucose, and the somatostatin analog octreotide, which alters gut transit and impairs release of vasoactive mediators into the bloodstream. PMID- 11879596 TI - Functional (Nonulcer) Dyspepsia. AB - Functional (nonulcer) dyspepsia refers to upper abdominal pain or discomfort with or without symptoms of early satiety, nausea, or vomiting with no definable organic cause. The current Rome II criteria help to diagnose functional dyspepsia and avoid misdiagnosis of gastroesophageal reflux disease and irritable bowel syndrome as functional dyspepsia. Assessment of gastric emptying with scintigraphy or breath testing may be useful in identifying delayed gastric emptying in patients with dyspeptic symptoms and may be helpful in patient management. Electrogastrography is a noninvasive test that evaluates for gastric dysrhythmias. Satiety testing is being evaluated as an indirect test for impaired fundic relaxation and visceral hypersensitivity. The symptom response to Helicobacter pylori therapy in patients with functional dyspepsia and a negative endoscopy examination but a positive H. pylori test is marginal. Lifestyle modifications often are suggested for initial treatment of functional dyspepsia. Dietary changes such as frequent small meals, low-fat diet, and avoidance of certain aggravating foods may improve symptoms. Additional measures include cessation of smoking, avoiding excess alcohol intake, and minimizing coffee intake. Antacids and over-the-counter histamine type 2 receptor antagonists may be helpful as an "on-demand" therapy for intermittent symptoms. They are safe and relatively inexpensive. Different subgroups of functional dyspepsia are based on the predominant symptom and may help in choosing an appropriate drug to initiate therapy. If the predominant symptom is epigastric pain (ulcer-like functional dyspepsia), histamine-2 receptor antagonists or proton pump inhibitors are the initial treatment of choice. If fullness, bloating, early satiety or nausea is the predominant complaint (dysmotility-like functional dyspepsia), a prokinetic agent may help. Metoclopramide is the only available effective prokinetic agent at present. If metoclopramide is used, short-term treatment and discussion of possible side effects with the patient are advised. If there is no response to these initial treatments, switching therapy from proton pump inhibitor to prokinetic or vice versa can be tried. If these treatment options fail, patient re-evaluation for other disorders (including other functional bowel disorders) is advised. A low-dose tricyclic antidepressant at bedtime may be helpful for treatment of visceral hypersensitivity. PMID- 11879595 TI - Acute Nonvariceal Upper Gastrointestinal Hemorrhage. AB - Upper endoscopy to assess the risk of rebleeding in patients with nonvariceal upper gastrointestinal bleeding may be used for triage, allowing outpatient care of selected patients and leading to significant cost savings. Over the last 10 years, hospitalization days required for upper gastrointestinal bleeding have decreased significantly and the majority of patients with upper gastrointestinal bleeding undergo endoscopy within 24 hours of admission. Twenty percent to 35% of these endoscopies include endoscopic hemostatic therapy. Endoscopic treatment is recommended for actively bleeding (ie, spurting or oozing) visible vessels and nonbleeding visible vessels that are raised and cannot be washed off. Endoscopic methods can be divided into thermal (multipolar coagulation, heater probe, argon plasma coagulator, Nd:YAG laser) and nonthermal (eg, injection therapy); both types are effective. A combination of injection and thermal therapy with initial injection to slow the bleeding or "clear the field" followed by coagulation of the identified vessel is popular. Bleeding recurs in 15% of patients. A recent randomized controlled trial of repeat endoscopic treatment versus surgery for patients with recurrent ulcer bleeding concluded that endoscopic retreatment is superior to surgery. Most peptic ulcer rebleeding occurs within the first 3 days of presentation. A comparison of omeprazole and placebo therapy in high-risk ulcer patients with bleeding stigmata at endoscopy who were not treated endoscopically found that high-dosage omeprazole (40 mg twice a day) significantly lowered the rates of further bleeding and surgical intervention. Although unlikely to replace endoscopic therapy, this study demonstrated the efficacy of potent acid suppression, perhaps due to stabilization of clotting activity. A recent placebo-controlled trial of high-dosage parenteral omeprazole after endoscopic treatment of bleeding peptic ulcers demonstrated a substantial reduction in the risk of rebleeding. PMID- 11879597 TI - [Proposals for adjusting the portfolio of services to today reality]. PMID- 11879598 TI - [Evolution of the communication profile of family medicine residents]. AB - AIMS: To study the development of patient relation skills, as used during interviews with patients for health problems that are common within their specialty, in family medicine residents during the third year of their residency program. METHODS: Quasi-experimental (before-after), national-level, multicenter study. The participants were 193 third-year residents in family medicine at 8 training units who were trained between 1996 and 1999. During this period all residents participated in the usual training and clinical activities included in the National Plan for this specialty. The GATHA-RES questionnaire was used to evaluated six clinical scenarios in video recordings of encounters with standardized patients (3 at the start of the third year and 3 at the end of the third year). Descriptive, bivariate and multivariate statistical analyses were used. RESULTS: A total of 1,024 interviews were analyzed. The time spent with each patient decreased significantly at the end of the residency program; the duration of the visit was directly proportional to the score on the GATHA-RES questionnaire (p < 0.05). Improvements were seen in formal and organizational aspects of the interview. In contrast, skills related with the exploration of personal and contextual aspects of the problem, and negotiating skills, were worse at the end of the study. The variables that best predicted residents' communicational profile were age (inverse relation), duration of the interview, training of the tutor in clinical interviewing, and teaching unit. CONCLUSIONS: Residents learn to shorten the duration of the visit to the detriment of communication skills that are basic to appropriate care for their patients' health problems. These results suggest the need for substantial changes in the training of family medicine residents in Spain. PMID- 11879599 TI - [To measure communication, that is the question]. PMID- 11879600 TI - [The social and economic cost of diabetes mellitus]. AB - OBJECTIVE: To evaluate the economic impact in terms of the direct costs (health care costs) and the indirect costs (effects on production caused by mortality and morbidity) of diabetes mellitus (DM) in the Canary Islands (Spain) during 1998. DESIGN: The approach used was to study the cost of the disease through looking only at people in the productive sphere (human capital). Direct and indirect costs were calculated by using costs of prevalence, i.e. the costs caused during 1998. SETTING: Direct costs were broken down into hospital care, primary care, out-patient consultations, drugs and further tests; indirect costs, into premature mortality and time off work. Measurements and main results. The total cost of DM amounted to 6468.76 million pesetas (38.88 millio ), or the equivalent of 126 168 pesetas (758.28 ) a year for each known diabetic patient. Total direct cost of DM was 4011.51 million pesetas (24.11 million ), which amounts to 2.13% of health expenditure in the Canaries; or the equivalent of 78 240 pesetas (470.23 ) a year for each known diabetic patient. Direct cost of health care was 62% of the total cost. Total indirect cost was 2457.25 million pesetas (14.77 million ), 38% of the total cost. CONCLUSIONS: Despite the conservatism of the approach, in that the study omitted costs linked to pain and suffering, permanent disability or home care by family members, the high social and economic cost of DM patients calls for cost-effectiveness studies that would permit a more transparent debate on this question. PMID- 11879602 TI - [Prevalence of anaemia in pregnancy and analysis of the underlying factors]. AB - OBJECTIVE: To determine the prevalence of anaemia and to evaluate the factors that condition its occurrence.Setting. Out-patient clinics in the La Plata area, Buenos Aires, Argentina. DESIGN: Observational and prospective study. PARTICIPANTS: All the pregnant women consulting for the first time, excluding those with prior pathology or regular use of medical drugs. MEASUREMENTS: Anaemia was defined at values of Hb < 11 g/dl. Questionnaires were administered for general data and the type of nutrition, and a complete haematological report was compiled. RESULTS: 1218 pregnant women started the study. Anaemia was detected in 196 of them (16%), with average Hb 9.88 g/dl. Between normal and anaemic pregnant women, the following differences were found between the first and second consultations: weight (64.44 vs 59.50, p < 0.00001), family income (US$744.36 vs 568.28, p < 0.0001), kilocalories ingested (2,488.44 vs 2,204.28, p = 0.01), percentage of proteins in diet (15.73 vs 13.69, p = 0.002), and weekly iron consumption (15.24 mg vs 13.04, p < 0.0001). CONCLUSIONS: Pregnant women run a greater risk of suffering anaemia if they have diets of < 1800 kcal, < 13% proteins, less than 7 mg of iron per week, and haemic iron < 10%; and family income below US$400. Ensuring a proper diet and improving the social and economic conditions of this population group will reduce the risk of anaemia during pregnancy and its perinatal consequences. PMID- 11879601 TI - [Cholesterol is still high. So what do we do now? Treatment of uncontrolled hypercholesterolaemia over a year]. AB - OBJECTIVE: To find the effectiveness of lipid-lowering treatment, based on normal clinical practice in primary care, on lipid control of patients with clear hypercholesterolaemia (HC). Design. Semi-experimental before-and-after intervention study. SETTING: Urban health centre. PARTICIPANTS: 187 patients known to have lipaemia, with total or LDL cCholesterol (cLDL) above 270 and 190 mg/dl, respectively.Intervention. Normal clinical practice for twelve months in nine primary care clinics. MAIN MEASUREMENTS: The lipid profile and lipid lowering treatment were recorded at the start of the study and after twelve months. Lipid control (as a function of cLDL) was evaluated as optimal, acceptable or deficient, as a function of the cardiovascular risk, following the criteria of the Spanish Arteriosclerosis Society (1994). RESULTS: In 27% of cases, no visit relating to HC was recorded by the patient s doctor. The number of patients treated with lipid-lowering drugs grew from 50 to 98 (27 vs 52%, p < 0,005), fundamentally at the expense of statin treatment. After twelve months, there were significant drops in the plasma concentration of cLDL (12%, 95%CI, 9 to 15%) and in the percentage of patients with deficient control, which fell from the initial 91% to 61% (p < 0.005), although only 16% reached optimal control. CONCLUSIONS: After a year, under conditions of normal clinical practice, there was an increase in the use of lipid-lowering drugs and improvement in lipid control, though a bit over half the patients (61%) with clear hypercholesterolaemia maintained concentrations requiring treatment. PMID- 11879605 TI - [Motivation and professional career in family medicine:Professional career and family doctors: reflections and proposal (II)]. PMID- 11879606 TI - [Ongoing medical training in Mexico]. PMID- 11879607 TI - [Bibliographic alert: how to keep up-to-date thanks to the Internet]. PMID- 11879608 TI - [Evaluation of a programme to use capillary blood to monitor orally taken anticoagulants in basic health areas far from the hospital]. PMID- 11879609 TI - [Oral contraceptives and non-alcoholic hepatic Steatosis]. PMID- 11879610 TI - [Primary care paediatricians and family doctors: rivals or colleagues?]. PMID- 11879612 TI - [Teaching: risk factor?]. PMID- 11879614 TI - [Antibiotics policy in the primary care field must not be the same as in hospital]. PMID- 11879615 TI - [67-Gallium citrate gammagraphy in the study of lymphomas]. PMID- 11879616 TI - [Effectiveness of (67)Ga scintigraphy in the diagnosis of lymphoma relapse]. AB - PURPOSE: To establish the clinical effectiveness of 67Ga in lymphoma recurrence. PATIENTS AND METHODS: Thirty-nine patients were assessed on 43 occasions (31 for Hodgkin's disease and 12 for non-Hodgkin's lymphoma) either for a suspected recurrence or to monitor the evolution of the disease. A computed tomography (CT) and a 67Ga whole body scan were performed. Independent observers who did not know the definitive diagnosis performed the CT and 67Ga readings. The gold standard was the biopsy results or the follow-up during a 12 months period. RESULTS: Recurrence was confirmed in 17 cases, while 26 continued to have complete remission. Using 67Ga resulted in true positive (TP), 24 true negative (TN), 2 false positive (FP) and 2 false negative (FN) results with a sensitivity of 0.88 (15/17), specificity of 0.92 (24/26), positive predictive value (PPV) of 0.88 (15/17) and negative predictive value (NPV) of 0.92 (24/26), while CT produced 12 TP, 20 TN, 6 FP and 5 FN, with a sensitivity of 0.7 (12/17), specificity of 0.77 (20/26), PPV of 0.88 (12/18) and NPV of 0.92 (20/25). CONCLUSIONS: 67Ga is a useful indicator and has better diagnostic effectiveness than CT for both the confirmation and exclusion of lymphoma recurrence. PMID- 11879617 TI - [Effect of different observers on the interpretation of pulmonary perfusion scintigraphy]. AB - AIM: The agreement in the interpretation of the scintigraphic images of pulmonary perfusion in the diagnosis of pulmonary tromboembolism (TEP) is not always the most adequate. The purpose of this study was to evaluate the degree of agreement by eight observers in the interpretation of these studies. MATERIAL AND METHODS: The study population consisted of 180 studies with clinically suspected pulmonary embolism referred to our department for scintigraphic perfusion imaging from April 98 to September 99. The patients received an intravenous injection of 111 148 MBq (3-4 mCi) of 99mTc-macroaggregated albumin whereas the images were obtained in the six routine projections. The observers consisted of five nuclear medicine physicians and three residents who independently reviewed the scintigrams (low, intermediate and high probability). In a second step, the physicians performed consensus interpretations. The Kappa statistics was used to evaluate the degree of agreement between individual observer interpretations beyond that expected by chance alone. RESULTS: The mean value SEM for Kappa index was 0.58 0.02, with maximum and minimum values of 0.76 and 0.27 respectively. The interobserver variability was greater than expected. Except in two observers scans in the interpretation of images in the low and high categories showed minor discrepancies, whereas scans in intermediate probability showed the most important interobserver variations. CONCLUSION: The study manifests the importance of a uniform criteria in lung scintigraphic interpretation. PMID- 11879618 TI - [Study of pulmonary lesions with (99m)Tc-Tetrafosmin and chest spect. Determination of uptake related factors, diagnostic value and prognosis]. AB - One hundred fifteen patients with 119 pulmonary lesions in which malignancy was suspected underwent a SPECT study with 99mTc-Tetrofosmin (TTF) to assess the possible factors involved in the uptake of the radiopharmaceutical. The TTF uptake rate in the lung tumor with respect to that of healthy tissue (TTF index) was evaluated in terms of: benignity and malignancy, histological type, stage, cell differentiation, size, necrosis, survival and the influence of P glycoprotein (Pgp), detected by immunohistochemistry, on the TTF uptake. The mean TTF index in the 18 benign lesions studied was 1.01 0.05, while that of the 101 malignant lesions was 1.59 0.45 (p < 0.001), with a positive predictive value of 97.7% and a negative predictive value of 50%. The comparison of the histological types, degree of cell differentiation, necrosis and stage revealed no statistically significant differences. With respect to size, those tumors measuring > 3 cm showed greater uptake than smaller lesions. In patients with non small cell lung cancer, a positive relationship was observed between the TTF index and survival, a circumstance that did not occur in patients with small cell lung cancer. In the cases in which the presence of Pgp was assessed, there was an inverse relationship between the TTF ratio and Pgp expression. In conclusion, thoracic SPECT with 99mTc-TTF has a high positive predictive value for the presence of lung cancer, although a negative study does not rule out the existence of the disease. The reason for this is the inverse relationship between 99mTc-TTF uptake and the density of Pgp expression. PMID- 11879620 TI - [Atypical findings of the combined scintigraphy of bone marrow and labeled leukocytes in osteonecrosis of the hip secondary to infection]. AB - Avascular osteonecrosis can be associated with septic arthritis and osteomyelitis. Combined labeled leukocyte-marrow imaging scintigraphy has demonstrated excellent accuracy for the detection of infection since both tracers accumulate in the bone marrow and only leukocytes accumulate in infection. We report an unusual total absence of 99mTc HMPAO leukocytes/9mTc-sulfur colloid tracer accumulation, not only in the femoral head but also in the acetabulum and hip in hip osteonecrosis secondary to septic arthritis and osteomyelitis. PMID- 11879619 TI - [Cytosolic pS2 levels in 154 non-small-cell lung carcinomas. Correlation with other clinical and biological parameters]. AB - INTRODUCTION: pS2 was first identified as an estrogen induced molecule in a breast cancer cell line, but its gene responds to several growth factors and oncogenic proteins. Its main clinical application lies in breast carcinomas, where it is a molecular marker to identify breast cancer patients who will respond to hormone therapy. This study was performed to evaluate the significance of the pS2 cytosolic levels in non small cell lung carcinomas classified according to different clinical and biological parameters. Likewise, the results obtained were correlated with those observed in normal tissues. MATERIAL AND METHODS: The study group included 154 non small cell lung carcinomas (59 adenocarcinomas and 95 squamous carcinomas) and 54 normal lung samples. Cathepsin D, pS2, neuron specific enolase (NSE) and CA125 cytosolic concentrations were determined, as well as those of epidermal growth factor receptor (EGFR), erbB2 protein, CD44s, CD44v5 and CD44v6 on cell surface membranes. Likewise, clinical stage, histological grade, ploidy and cellular S-phase fraction were also considered as variables of the study. RESULTS: We observed cytosolic pS2 values greater than 1 ng/mg protein (positive cutoff) in 18/59 adenocarcinomas and in 10/95 squamous carcinomas (p:0.00177). In adenocarcinomas, pS2 positivity was not correlated with any clinical and biological parameters, but positive cases had lower EGFR (p:0.03770) concentrations and higher CA125 (p:0.01902) levels. In squamous carcinomas, pS2 positivity was associated with lower EGFR (p:0.02270) levels and higher erbB2 protein (p:0,00563) concentrations. CONCLUSIONS: Our results led us to suggest the following: 1) cytosolic pS2 concentrations are higher in tumoral than in normal samples; 2) adenoarcinomas had higher levels than squamous lung carcinomas; 3) in adenocarcinomas, positivity for pS2 is associated with lower EGFR levels and higher CA125 concentrations suggesting a worse outcome, and 4) in squamous lung tumors, positivity for pS2 was associated with lower EGFR and higher erbB2 oncoprotein concentrations, it not being possible to establish its clinical significance, although it may be correlated with a poor prognosis. PMID- 11879621 TI - [Whole-body (67)Ga scintigraphy in a patient with thymus carcinoma]. AB - We report a case of a 50 year old man referred to our department with a history of mild cough, dyspnea and dysphagia. The thoracic CT scan showed a large solid mass in the anterior mediastinum, corresponding to the findings in the chest radiographs.A 67Ga scintigraphy was performed and showed high pathological accumulation in the anterior mediastinum. A subsequent fine needle aspiration biopsy (FNAB) showed the presence of malignant cells, suggesting thymic carcinoma. Although this type of tumour is uncommon, it should be taken into account in order to establish the differential diagnosis of gallium-avid mediastinal masses. PMID- 11879622 TI - [Isotopic scan with somatostatin receptors in a case of recurrent carcinoid tumor. Relevance of tomographic detection]. PMID- 11879623 TI - [Breast osteosarcoma]. PMID- 11879624 TI - [On the isotopic localization of parathyroid adenomas]. PMID- 11879625 TI - [Whole body positron emission tomography (PET) with (18F)-fluorodeoxyglucose]. PMID- 11879626 TI - [Positron emission tomography (PET) in clinical oncology. Part II]. PMID- 11879627 TI - Oxidants painting the cysteine chapel: redox regulation of PTPs. AB - Growth factors and cytokines appear to stimulate the intracellular production of reactive oxygen species (ROS). Evidence suggests that this alteration in the cellular redox state is essential for downstream signaling, but the precise mechanism has remained elusive. A new study now demonstrates that ligand stimulated intracellular hydrogen peroxide can specifically and reversibly regulate the activity of protein tyrosine phosphatases. PMID- 11879628 TI - Photoreceptor cells in flies and mammals: Crumby homology? AB - Recently, two papers have revealed a new function for the fruit fly epithelial apical membrane protein Crumbs and its mammalian homolog CRB1 in photoreceptor cell morphogenesis. This supports the previous observation that disruption of CRB1 function can cause retinal degeneration in humans. PMID- 11879629 TI - The "Down's" side of mitochondria. AB - The mechanisms involved in the development of Alzheimer disease-type pathology in patients with Down's syndrome are unclear. However, a paper published in the February 28 issue of Neuron indicates that mitochondrial dysfunction is pivotally involved. PMID- 11879630 TI - Apoptosome: the seven-spoked death machine. AB - A paper in the February issue of Molecular Cell describes the structure of the apoptosome, a multicomponent death machine, deciphered by cryoelectron microscopy. This structure explains the assembly of the machine, the tentative location of the subcomponents, and proposes a mechanism for initiating the death signal. PMID- 11879631 TI - Fusion of vacuoles--where are the membranes, and where are the holes? AB - In the February 8th issue of Cell, Wang et al. report the surprising finding that vacuolar fusion occurs at the periphery of the contact area of the vacuoles and not by the expansion of a central fusion pore. During fusion, a disk of boundary membrane is excised and left behind within the fused vacuoles. PMID- 11879632 TI - Protein and RNA export from the nucleus. AB - The presence of the nuclear envelope necessitates the movement of proteins and RNAs between the nucleus and the cytoplasm. Elaborate cellular machinery exists to promote the nuclear transport of macromolecules. Recent advances in the field have illuminated our comprehension of both nuclear import and export as powerful means of gene regulation. As our appreciation of the importance of the process has grown, its study has matured, moving beyond the single cell to the entire organism. This review discusses basic mechanisms and regulation of protein, mRNA, and ribosome export with an emphasis on developmental examples. PMID- 11879633 TI - Plotting a course: multiple signals guide pollen tubes to their targets. AB - Pollen plays a critical role in the life cycle of all flowering plants, generating a polarized pollen tube that delivers sperm to the eggs in the interior of the flower. Pollen tubes perceive multiple extracellular signals during their extended growth through different floral environments; these environments discriminate among pollen grains, allowing only those that are appropriately recognized to invade. The phases of pollen tube growth include interactions that establish pollen polarity, entry of pollen tubes into female cell walls, and adhesion-based pollen tube motility through a carbohydrate-rich matrix. Recent studies have identified cells within the female germ unit as important sources of pollen guidance cues. Other signals undoubtedly exist, and their discovery will require genetic screens that target diploid tissues as well as haploid male and female cells. PMID- 11879634 TI - The yeast clathrin adaptor protein complex 1 is required for the efficient retention of a subset of late Golgi membrane proteins. AB - In yeast, certain resident trans-Golgi network (TGN) proteins achieve steady state localization by cycling through late endosomes. Here, we show that chitin synthase III (Chs3p), an enzyme involved in the assembly of the cell wall at the mother-bud junction, populates an intracellular reservoir that is maintained by a cycle of transport between the TGN and early endosomes. Traffic of Chs3p from the TGN/early endosome to the cell surface requires CHS5 and CHS6, mutant alleles of which trap Chs3p in the TGN/early endosome. Disruption of the clathrin adaptor protein complex 1 (AP-1) restores Chs3p transport to the plasma membrane. Similarly, in AP-1 deficient cells, the resident TGN/early endosome syntaxin, Tlg1p, is missorted. We propose that clathrin and AP-1 act to recycle Chs3p and Tlg1p from the early endosome to the TGN. PMID- 11879635 TI - Sly1 binds to Golgi and ER syntaxins via a conserved N-terminal peptide motif. AB - Sec1/munc18-like proteins (SM proteins) and SNARE complexes are probably universally required for membrane fusion. However, the molecular mechanism by which they interact has only been defined for synaptic vesicle fusion where munc18 binds to syntaxin in a closed conformation that is incompatible with SNARE complex assembly. We now show that Sly1, an SM protein involved in Golgi and ER fusion, binds to a short, evolutionarily conserved N-terminal peptide of Sed5p and Ufe1p in yeast and of syntaxins 5 and 18 in vertebrates. In these syntaxins, the Sly1 binding peptide is upstream of a separate, autonomously folded N terminal domain. These data suggest a potentially general mechanism by which SM proteins could interact with peptides in target proteins independent of core complex assembly and suggest that munc18 binding to syntaxin is an exception. PMID- 11879636 TI - The Rab GTPase Ypt1p and tethering factors couple protein sorting at the ER to vesicle targeting to the Golgi apparatus. AB - GPI-anchored proteins exit the ER in distinct vesicles from other secretory proteins, and this sorting event can be reproduced in vitro. When extracts from a uso1 mutant were used, the sorting of GPI-anchored proteins from other secretory proteins was defective. Complementation with purified Uso1p restored sorting. The Rab GTPase Ypt1p and the tethering factors Sec34p and Sec35p, but not Bet3p, a member of the TRAPP complex, were also required for protein sorting upon ER exit. Therefore, the Ypt1p tethering complex couples protein sorting in the ER to vesicle targeting to the Golgi apparatus. Sorting of GPI-anchored proteins from other secretory proteins was also observed in vivo. The sorting defect observed in vitro with uso1 and ypt1 mutants was reproduced in vivo. PMID- 11879637 TI - Conserved organization of centromeric chromatin in flies and humans. AB - Recent studies have highlighted the importance of centromere-specific histone H3 like (CENP-A) proteins in centromere function. We show that Drosophila CID and human CENP-A appear at metaphase as a three-dimensional structure that lacks histone H3. However, blocks of CID/CENP-A and H3 nucleosomes are linearly interspersed on extended chromatin fibers, and CID is close to H3 nucleosomes in polynucleosomal preparations. When CID is depleted by RNAi, it is replaced by H3, demonstrating flexibility of centromeric chromatin organization. Finally, contrary to models proposing that H3 and CID/CENP-A nucleosomes are replicated at different times in S phase, we show that interspersed H3 and CID/CENP-A chromatin are replicated concurrently during S phase in humans and flies. We propose that the unique structural arrangement of CID/CENP-A and H3 nucleosomes presents centromeric chromatin to the poleward face of the condensing mitotic chromosome. PMID- 11879638 TI - Inhibition of PPAR gamma 2 gene expression by the HIF-1-regulated gene DEC1/Stra13: a mechanism for regulation of adipogenesis by hypoxia. AB - Cellular differentiation involves transcriptional responses to environmental stimuli. Adipocyte differentiation is inhibited under hypoxic conditions, indicating that oxygen (O(2)) is an important physiological regulator of adipogenesis. Hypoxia inhibits PPAR gamma 2 nuclear hormone receptor transcription, and overexpression of PPAR gamma 2 or C/EBP beta stimulates adipogenesis under hypoxia. Mouse embryonic fibroblasts deficient in hypoxia inducible transcription factor 1 alpha (HIF-1 alpha) are refractory to hypoxia mediated inhibition of adipogenesis. The HIF-1-regulated gene DEC1/Stra13, a member of the Drosophila hairy/Enhancer of split transcription repressor family, represses PPAR gamma 2 promoter activation and functions as an effector of hypoxia-mediated inhibition of adipogenesis. These data indicate that an O(2) sensitive signaling mechanism regulates adipogenesis. Thus, agents that regulate HIF-1 activity or O(2) sensing may be used to inhibit adipogenesis and control obesity. PMID- 11879639 TI - Half pint regulates alternative splice site selection in Drosophila. AB - Alternative splicing is used by metazoans to increase protein diversity and to alter gene expression during development. However, few factors that control splice site choice in vivo have been identified. Here we describe a factor, Half pint (Hfp), that regulates RNA splicing in Drosophila. Females harboring hypomorphic mutations in hfp lay short eggs and show defects in germline mitosis, nuclear morphology, and RNA localization during oogenesis. We find that hfp encodes the Drosophila ortholog of human PUF60 and functions in both constitutive and alternative splicing in vivo. In particular, hfp mutants display striking defects in the developmentally regulated splicing of ovarian tumor (otu). Furthermore, transgenic expression of the missing otu splice form can rescue the ovarian phenotypes of hfp. PMID- 11879640 TI - The type I membrane protein EFF-1 is essential for developmental cell fusion. AB - Multinucleate cells are widespread in nature, yet the mechanism by which cells fuse their plasma membranes is poorly understood. To identify animal fusogens, we performed new screens for mutations that abolish cell fusion within tissues of C. elegans throughout development. We identified the gene eff-1, which is expressed as cells acquire fusion competence and encodes a novel integral membrane protein. EFF-1 sequence motifs suggest physicochemical actions that could cause adjacent bilayers to fuse. Mutations in the extracellular domain of EFF-1 completely block epithelial cell membrane fusion without affecting other perfusion events such as cell generation, patterning, differentiation, and adhesion. Thus, EFF-1 is a key component in the mechanism of cell fusion, a process essential to normal animal development. PMID- 11879641 TI - Stat3 Controls Cell Movements during Zebrafish Gastrulation. AB - Vertebrate axis formation requires both the correct specification of cell fates and the coordination of gastrulation movements. We report that the zebrafish signal transducer and activator of transcription 3 (Stat3) is activated on the dorsal side by the maternal Wnt/beta-catenin pathway. Zebrafish embryos lacking Stat3 activity display abnormal cell movements during gastrulation, resulting in a mispositioned head and a shortened anterior-posterior axis, but show no defects in early cell fate specification. Time course analysis, cell tracing, and transplantation experiments revealed that Stat3 activity is required cell autonomously for the anterior migration of dorsal mesendodermal cells and non cell autonomously for the convergence of neighboring paraxial cells. These results reveal a role for Stat3 in controlling cell movements during gastrulation. PMID- 11879642 TI - Synaptic plasticity in an altered state. AB - In this issue of Neuron, record from synaptically coupled pairs of CA3 neurons to closely examine the induction of synaptic depression at a small number of identified synapses. The authors provide convincing evidence that the activation history of a synapse determines both the ability of a synapse to depress and the mechanism of depression. PMID- 11879643 TI - The nature of illusory contour computation. AB - Neural correlates of illusory contour perception have been found in both the early and the higher visual areas. But the locus and the mechanism for its computation remain elusive. Psychophysical evidence provided in this issue of Neuron shows that perceptual contour completion is likely done in the early visual cortex in a cascade manner using horizontal connections. PMID- 11879644 TI - Toward an understanding of the brain substrates of reward in humans. AB - A network of brain regions has been implicated in food-reward processing. now provide evidence that this network is differentially modulated by anticipation versus receipt of a food reward and suggest an additional effect of valence of the stimulus. PMID- 11879645 TI - Genetic models meet trophic mechanisms: EGF family members are gliatrophins in Drosophila. AB - Trophic survival mechanisms are crucial for the determination of cell numbers in the developing vertebrate nervous system, but important neurotrophic factor families such as the neurotrophins have not yet been found in either Drosophila or C. elegans. Two independent studies on distinct glial populations in Drosophila have now shown that their survival is regulated by EGF family members secreted by adjacent neurons. Fly genetics thus promises new insights on trophic signaling mechanisms and confirms that trophic regulation of cell survival is an evolutionarily ancient mechanism for building the nervous system. PMID- 11879646 TI - Altered metabolism of the amyloid beta precursor protein is associated with mitochondrial dysfunction in Down's syndrome. AB - Most Down's syndrome (DS) patients develop Alzheimer's disease (AD) neuropathology. Astrocyte and neuronal cultures derived from fetal DS brain show alterations in the processing of amyloid beta precursor protein (AbetaPP), including increased levels of AbetaPP and C99, reduced levels of secreted AbetaPP (AbetaPPs) and C83, and intracellular accumulation of insoluble Abeta42. This pattern of AbetaPP processing is recapitulated in normal astrocytes by inhibition of mitochondrial metabolism, consistent with impaired mitochondrial function in DS astrocytes. Intracellular Abeta42 and reduced AbetaPPs are also detected in DS and AD brains. The survival of DS neurons is markedly increased by recombinant or astrocyte-produced AbetaPPs, suggesting that AbetaPPs may be a neuronal survival factor. Thus, mitochondrial dysfunction in DS may lead to intracellular deposition of Abeta42, reduced levels of AbetaPPs, and a chronic state of increased neuronal vulnerability. PMID- 11879647 TI - Retinal ganglion cells do not extend axons by default: promotion by neurotrophic signaling and electrical activity. AB - We investigate the signaling mechanisms that induce retinal ganglion cell (RGC) axon elongation by asking whether surviving neurons extend axons by default. We show that bcl-2 overexpression is sufficient to keep purified RGCs alive in the absence of any glial or trophic support. The bcl-2-expressing RGCs do not extend axons or dendrites unless signaled to do so by single peptide trophic factors. Axon growth stimulated by peptide trophic factors is remarkably slow but is profoundly potentiated by physiological levels of electrical activity spontaneously generated within embryonic explants or mimicked on a multielectrode silicon chip. These findings demonstrate that these surviving neurons do not constitutively extend axons and provide insight into the signals that may be necessary to promote CNS regeneration. PMID- 11879648 TI - Dissecting intersubunit contacts in cyclic nucleotide-gated ion channels. AB - In cyclic nucleotide-gated (CNG) ion channels, binding of cGMP or cAMP drives a conformational change that leads to opening of an ion-conducting pore. One region implicated in the coupling of ligand binding to opening of the pore is the C linker region. Here, we used crosslinking of endogenous cysteines to study interregion proximity. We demonstrate that an individual amino acid--C481--in the C linker region of each of two neighboring subunits can form a disulfide bond. Further, using tandem dimers, we show that a disulfide bond between C35 in the N terminal region and C481 in the C linker region can form either within a subunit or between subunits. From our data on proximity between individual amino acids and previous studies, a picture emerges of the C linker as a potential dimerization interface. PMID- 11879649 TI - Diverse trafficking patterns due to multiple traffic motifs in G protein activated inwardly rectifying potassium channels from brain and heart. AB - G protein-activated inwardly rectifying potassium channels (Kir3, GIRK) provide an important mechanism for neurotransmitter regulation of membrane excitability. GIRK channels are tetramers containing various combinations of Kir3 subunits (Kir3.1--Kir3.4). We find that different combinations of Kir3 subunits exhibit a surprisingly complex spectrum of trafficking phenotypes. Kir3.2 and Kir3.4, but not Kir3.1, contain ER export signals that are important for plasma membrane expression of Kir3.1/Kir3.2 and Kir3.1/Kir3.4 heterotetramers, the GIRK channels found in the brain and the heart, respectively. Additional motifs in Kir3.2 and Kir3.4 control the trafficking between endosome and plasma membrane. In contrast, the Kir3.3 subunit potently inhibits plasma membrane expression by diverting the heterotetrameric channels to lysosomes. Such rich trafficking behaviors provide a mechanism for dynamic regulation of GIRK channel density in the plasma membrane. PMID- 11879650 TI - 3-phosphoinositides modulate cyclic nucleotide signaling in olfactory receptor neurons. AB - Phosphatidylinositol 3-kinase (PI3K)-dependent phosphoinositide signaling has been implicated in diverse cellular systems coupled to receptors for many different ligands, but the extent to which it functions in sensory transduction is yet to be determined. We now report that blocking PI3K activity increases odorant-evoked, cyclic nucleotide-dependent elevation of [Ca(2+)](i) in acutely dissociated rat olfactory receptor neurons and does so in an odorant-specific manner. These findings imply that 3-phosphoinositide signaling acts in vertebrate olfactory transduction to inhibit cyclic nucleotide-dependent excitation of the cells and that the interaction of the two signaling pathways is important in odorant coding, indicating that 3-phosphoinositide signaling can play a role in sensory transduction. PMID- 11879651 TI - Regulation of rho GTPases by crosstalk and neuronal activity in vivo. AB - Proper development of neurons depends on synaptic activity, but the mechanisms of activity-dependent neuronal growth are not well understood. The small GTPases, RhoA, Rac, and Cdc42, regulate neuronal morphogenesis by controlling the assembly and stability of the actin cytoskeleton. We report an in situ method to determine endogenous Rho GTPase activity in intact Xenopus brain. We use this method to provide evidence for crosstalk between Rho GTPases in optic tectal cells. Moreover, crosstalk between the Rho GTPases appears to affect dendritic arbor development in vivo. Finally, we demonstrate that optic nerve stimulation regulates Rho GTPase activity in a glutamate receptor-dependent manner. These data suggest a link between glutamate receptor function, Rho GTPase activity, and dendritic arbor growth in the intact animal. PMID- 11879652 TI - Negative regulation and gain control of sensory neurons by the C. elegans calcineurin TAX-6. AB - Animals sense and adapt to variable environments by regulating appropriate sensory signal transduction pathways. Here, we show that calcineurin plays a key role in regulating the gain of sensory neuron responsiveness across multiple modalities. C. elegans animals bearing a loss-of-function mutation in TAX-6, a calcineurin A subunit, exhibit pleiotropic abnormalities, including many aberrant sensory behaviors. The tax-6 mutant defect in thermosensation is consistent with hyperactivation of the AFD thermosensory neurons. Conversely, constitutive activation of TAX-6 causes a behavioral phenotype consistent with inactivation of AFD neurons. In olfactory neurons, the impaired olfactory response of tax-6 mutants to an AWC-sensed odorant is caused by hyperadaptation, which is suppressible by a mutation causing defective olfactory adaptation. Taken together, our results suggest that stimulus-evoked calcium entry activates calcineurin, which in turn negatively regulates multiple aspects of sensory signaling. PMID- 11879653 TI - State-dependent heterogeneity in synaptic depression between pyramidal cell pairs. AB - Paired recordings between CA3 pyramidal neurons were used to study the properties of synaptic plasticity in active and silent synapses. Synaptic depression is accompanied by decreases in both AMPAR and NMDAR function. The mechanisms of synaptic depression, and the potential to undergo activity-dependent plastic changes in efficacy, differ depending on whether a synapse is active, recently silent, or potentiated. These results suggest that silent and active synapses represent distinct synaptic "states," and that once unsilenced, synapses express plasticity in a graded manner. The state in which a synapse resides, and the states recently visited, determine its potential and mechanism for undergoing subsequent plastic changes. PMID- 11879654 TI - Contributions of receptor desensitization and saturation to plasticity at the retinogeniculate synapse. AB - The retinogeniculate synapse conveys visual information from the retina to thalamic relay neurons. Here, we examine the mechanisms of short-term plasticity that can influence transmission at this connection in mouse brain slices. Our studies show that synaptic strength is modified by physiological activity patterns due to marked depression at high frequencies. Postsynaptic mechanisms of plasticity make prominent contributions to this synaptic depression. During trains of retinal input stimulation, receptor desensitization attenuates the AMPA EPSC while the NMDA EPSC saturates. This differential plasticity may help explain the distinct roles of these receptors in shaping the relay neuron response to visual stimulation with the AMPA component being important for transient responses, while sustained high frequency responses rely more on the NMDA component. PMID- 11879655 TI - Decreased synaptic vesicle recycling efficiency and cognitive deficits in amphiphysin 1 knockout mice. AB - The function of the clathrin coat in synaptic vesicle endocytosis is assisted by a variety of accessory factors, among which amphiphysin (amphiphysin 1 and 2) is one of the best characterized. A putative endocytic function of amphiphysin was supported by dominant-negative interference studies. We have now generated amphiphysin 1 knockout mice and found that lack of amphiphysin 1 causes a parallel dramatic reduction of amphiphysin 2 selectively in brain. Cell-free assembly of endocytic protein scaffolds is defective in mutant brain extracts. Knockout mice exhibit defects in synaptic vesicle recycling that are unmasked by stimulation and suggest impairments at multiple stages of the cycle. These defects correlate with increased mortality due to rare irreversible seizures and with major learning deficits, suggesting a critical role of amphiphysin for higher brain functions. PMID- 11879656 TI - Perceptual completion across the vertical meridian and the role of early visual cortex. AB - Perceptual completion can link widely separated contour fragments and interpolate illusory contours (ICs) between them. The mechanisms underlying such long-range linking are not well understood. Here we report that completion is much poorer when ICs cross the vertical meridian than when they reside entirely within the left or right visual hemifield. This deficit reflects limitations in cross hemispheric integration. We also show that the sensitivity to the interhemispheric divide is unique to perceptual completion: a comparable task which did not require completion showed no across-meridian impairment. We propose that these findings support the existence of specialized completion mechanisms in early visual cortical areas (V1/V2), since those areas are likely to be more sensitive to the interhemispheric divide. PMID- 11879657 TI - Neural responses during anticipation of a primary taste reward. AB - The aim of this study was to determine the brain regions involved in anticipation of a primary taste reward and to compare these regions to those responding to the receipt of a taste reward. Using fMRI, we scanned human subjects who were presented with visual cues that signaled subsequent reinforcement with a pleasant sweet taste (1 M glucose), a moderately unpleasant salt taste (0.2 M saline), or a neutral taste. Expectation of a pleasant taste produced activation in dopaminergic midbrain, posterior dorsal amygdala, striatum, and orbitofrontal cortex (OFC). Apart from OFC, these regions were not activated by reward receipt. The findings indicate that when rewards are predictable, brain regions recruited during expectation are, in part, dissociable from areas responding to reward receipt. PMID- 11879658 TI - Under-recruitment and nonselective recruitment: dissociable neural mechanisms associated with aging. AB - Frontal contributions to cognitive decline in aging were explored using functional MRI. Frontal regions active in younger adults during self-initiated (intentional) memory encoding were under-recruited in older adults. Older adults showed less activity in anterior-ventral regions associated with controlled use of semantic information. Under-recruitment was reversed by requiring semantic elaboration suggesting it stemmed from difficulty in spontaneous recruitment of available frontal resources. In addition, older adults recruited multiple frontal regions in a nonselective manner for both verbal and nonverbal materials. Lack of selectivity was not reversed during semantically directed encoding even when under-recruitment was diminished. These findings suggest two separate forms of age-associated change in frontal cortex: under-recruitment and nonselective recruitment. The former is reversible and potentially amenable to cognitive training; the latter may reflect a less malleable change associated with cognitive decline in advanced aging. PMID- 11879659 TI - Documenting regression of atherosclerosis: practical approaches in drug development. PMID- 11879660 TI - Quantitative coronary angiography in regression trials: a review of methodologic considerations, endpoint selection, and limitations. AB - Several quantitative angiographic methods (visual assessment, digital hand-held calipers, and automated edge detection) have been successfully used to assess coronary atherosclerosis regression. These methods are applicable for (1) continuous measures of disease progression and regression, including patient based, segment-based, and lesion-based analyses; and (2) categorical assessment of disease progression and regression (the use of a predefined threshold to categorize the course of disease). Angiographic methods and criteria for assessing disease progression each have distinct advantages and disadvantages, which accounts for the methodologic diversity seen in clinical trials. PMID- 11879661 TI - Ultrasound measurement of carotid plaque as a surrogate outcome for coronary artery disease. AB - Intima-media thickness (IMT) is commonly used as a surrogate marker for coronary artery disease (CAD). However, use of this parameter is a problem because (1) it assumes uniform thickness throughout the blood vessel, and (2) it detects changes primarily in the media, whereas atherosclerosis is a focal phenomenon that is confined to the intima. As an alternative to IMT, we have investigated the use of ultrasound measurements of plaque area and plaque volume as surrogate outcomes for CAD. Plaque area is a sensitive predictor of coronary disease progression and is closely associated with CAD. Clinical studies have shown that the sample size needed to detect atherosclerosis progression is sufficiently small to make this a useful assessment in clinical trials. Plaque volume, as measured by 3-dimensional ultrasound, may offer an even better means of assessing atherosclerosis. Plaque volume assessments are highly accurate and the data can be saved on a compact disc for central reading. Because of the high degree of accuracy and the increased size of plaque volume relative to plaque area or IMT, it is likely that only a small sample size will be required to detect clinically meaningful differences in plaque volume in a clinical trial. PMID- 11879663 TI - Usefulness of electron-beam computed tomography. AB - The field of atherosclerosis imaging has expanded rapidly in the last decade, and technologies such as electron-beam computed tomography (EBCT), have contributed significantly to our understanding of the prevalence of silent coronary artery disease and its consequences. Nonetheless, proper use of technology is necessary to conduct effective and cost-beneficial screening programs. Because most adverse events related to atherosclerosis occur in individuals at an intermediate risk level, it seems appropriate to concentrate screening efforts on this group of patients. This article reviews the current understanding of the value of coronary artery calcium screening in asymptomatic individuals and in symptomatic patients at low-to-intermediate risk and the use of EBCT as a tool for assessing the efficacy of therapy for atherosclerosis. PMID- 11879665 TI - Application of intravascular ultrasound to characterize coronary artery disease and assess the progression or regression of atherosclerosis. AB - Angiography has major limitations in its ability to assess coronary disease. Intravascular ultrasound (IVUS) offers unique capabilities to assess coronary atherosclerotic burden. The tomographic orientation of ultrasound enables visualization of the full vessel wall, as opposed to the 2-dimensional projection of the lumen provided by angiography. The equipment required to perform coronary IVUS consists of a catheter with a miniaturized transducer and a console to reconstruct the image. High ultrasound frequencies are used, typically, 30 to 40 MHz, which provides excellent theoretical resolution. IVUS has been performed safely in a wide variety of clinical situations. Vessels with classic atherosclerosis exhibit a diversity of abnormal features that reflect the severity, composition, and distribution of the atheromata. Plaque rupture is sometimes evident in ultrasound examination of the culprit lesions after an acute coronary syndrome. Most laboratories routinely perform cross-sectional area measurements of the lumen and external elastic membrane boundaries and calculate atheroma area. IVUS commonly detects atherosclerosis at angiographically normal sites. It has contributed substantially to our understanding of remodeling and has shown that positive remodeling is more prevalent in unstable lesions. Studies in patients early after transplantation have shown the presence of advanced atherosclerosis in their apparently normal donors. In addition, the application of IVUS in detecting the rate of progression or regression of existing atherosclerosis is among the most dynamic areas of development. IVUS is likely to emerge as the "gold standard" in the study of atherosclerosis progression regression over the next few years. PMID- 11879668 TI - A discussion of modalities for assessing regression and progression in vascular disease. PMID- 11879666 TI - An update on carotid ultrasound measurement of intima-media thickness. AB - Coronary atherosclerosis is a chronic, multifactorial disease process. Some individuals with atherosclerosis receive treatment in the form of mechanical or pharmacologic interventions after an acute event has occurred. Others receive treatment in the form of risk factor-based systemic intervention, the effectiveness of which is assessed by its ability to prevent an acute event. Surrogate endpoints in the study of atherosclerosis interventions are needed to better define disease course and disease response to interventions during the prolonged asymptomatic period. Several techniques for assessing arterial health are available, including quantitative intima-media thickness (QIMT) measurement by carotid ultrasound. QIMT is a safe, validated, and portable method that may prove highly useful in screening for atherosclerosis and in providing a surrogate measure for response to disease interventions. PMID- 11879669 TI - Adrenomedullin and the microvasculature. PMID- 11879671 TI - Microcirculation down under. AB - The Seventh World Congress for Microcirculation, organized by the Australian and New Zealand Microcirculation Society, was held on 19-22 August 2001 in Sydney, New South Wales, Australia. PMID- 11879670 TI - PTEN: a new twist on beta-catenin? PMID- 11879672 TI - Is there a novel brain interleukin 1 receptor that signals ischaemic brain damage? PMID- 11879673 TI - Pro-inflammatory mechanisms of a non-steroidal anti-inflammatory drug. PMID- 11879675 TI - Resisting resistance with PAF receptor antagonists. PMID- 11879676 TI - Don't forget Buchheim. PMID- 11879677 TI - Current status and future directions in the pharmacotherapy of epilepsy. AB - Considerable progress in the pharmacotherapy of epilepsy has been witnessed during the 20th century. However, despite the development of various antiepileptic drugs, about a third of patients are resistant to current pharmacotherapies. Even in patients in whom pharmacotherapy is efficacious, current antiepileptic drugs do not affect the progression or underlying natural history of the condition. Furthermore, currently there are no drugs available that prevent the development of epilepsy following, for example, head trauma. The rapid expansion of information about the cellular, molecular and genetic mechanisms of epilepsy is expected to lead to more effective therapies, prevention or even a cure for different types of epilepsy. In this article, I assess the current status of antiepileptic therapies and highlight innovative approaches for future treatments. PMID- 11879678 TI - 'Biogenerics': the off-patent biotech products. AB - The first patents of biopharmaceuticals derived from recombinant DNA will expire shortly, which raises the possibility of marketing generic products ('biogenerics') with limited documentation, similar to that which occurs with conventional pharmaceuticals. We propose the term off-patent biotechnological products (OPBPs) as an alternative to biogenerics when describing such products. It is questionable whether the majority of OPBPs can be classified as similar to the innovator products, considering the size and complexity of the molecules and the many factors that influence biological activity. There are three classes of OPBPs, each of which needs to meet different regulatory demands when seeking marketing authorization. PMID- 11879679 TI - Poly(ADP-ribose) polymerase: killer or conspirator? The 'suicide hypothesis' revisited. AB - Poly(ADP-ribose) polymerase 1 (PARP-1) is an abundant nuclear enzyme involved in DNA repair. The therapeutic efficacy of drugs that inhibit PARP-1 in various disorders underscores the active role of PARP-1 in cell death. Although it is well established that excessive DNA damage causes PARP-1 hyperactivation, which leads to cell death by energy failure, a new mechanistic perspective is emerging following the identification of various PARPs that exhibit different features and subcellular distributions. Studies demonstrating the significant role of PARP-1 in the regulation of gene transcription have further increased the intricacy of poly(ADP-ribosyl)ation in the control of cell homeostasis and challenge the notion that energy collapse is the sole mechanism by which poly(ADP-ribose) formation contributes to cell death. The hypothesis that PARPs might regulate cell fate as essential modulators of death and survival transcriptional programs will be discussed with particular focus on the regulation of transcription factors such as nuclear factor kappaB and p53. (An animation depicting the involvement of PARP-1 in the 'suicide hypothesis' is available at http://archive.bmn.com/supp/tips/tips2303a.html) PMID- 11879680 TI - Nicotine addiction: the possible role of functional upregulation. AB - Upregulation of binding to nicotinic acetylcholine receptors (nAChRs) is observed in the brains of both smokers and animals chronically exposed to nicotine, although whether this in vivo change is accompanied by an increase in receptor function is unknown. In vitro recordings indicate that alpha4beta2- and alpha7 subtypes of nAChRs, which are the most abundant subtypes in the brain, are functionally upregulated following prolonged exposure to nicotine. The possible consequences of functional upregulation for nicotine addiction are discussed. Moreover, we propose a new paradigm that describes the unusual behavior of these neuronal nAChRs and helps to explain the effects of nicotine in the CNS and the diffuse effects of ACh. PMID- 11879681 TI - Toxic heavy metals and undeclared drugs in Asian herbal medicines. AB - Asian herbal medicines are currently used by large sections of the population. Because they are not regulated as medicines and are freely available to everyone, serious safety concerns might be associated with these herbal medicines. In this article, evidence suggesting that some Asian herbal medicines contain toxic heavy metals or undeclared prescription drugs is reviewed. In particular, Indian and Chinese preparations have been implicated. Although adulteration with drugs is by definition fraudulent, the inclusion of heavy metals could be either intentional for alleged medicinal purposes or accidental. Evidence from various countries implies that toxic heavy metals and undeclared prescription drugs in Asian herbal medicines might constitute a serious health problem. However, the majority of the data is anecdotal and insufficient to define prevalence figures. Ways ought to be found to maximize consumer safety. PMID- 11879682 TI - Seven-transmembrane receptors: crystals clarify. AB - The X-ray structure of the photoreceptor rhodopsin has provided the first atomic resolution structure of a seven-transmembrane (7-TM) G-protein-coupled receptor. This has provided an improved template for interpreting the huge body of structure--activity, mutagenesis and affinity labelling data available for related 7-TM receptors, such as muscarinic acetylcholine receptors. Ligand contacts, and the intramolecular interactions that stabilize the ground state structure, can be identified with some degree of confidence. We now have a firm basis for attempts to predict the structure of the receptor--G-protein complex, and understand the mechanism by which the agonist--receptor complex activates the G protein. PMID- 11879687 TI - Comparison and verification of quantitative competitive reverse transcription polymerase chain reaction (QC-RT-PCR) and real time RT-PCR for avian leukosis virus subgroup J. AB - Avian leukosis virus subgroup J (ALV-J) infections cause significant economic losses because of increased mortality, tumor production, decreased production, and cost for eradication. Current quantification methods for ALV-J expressed by TCID(50) are difficult to determine because of the lack of cytopathic effect in cell cultures and non-specificity of currently available antigen-capture ELISA tests. In this study, a one-tube fluorescent probe based real time RT-PCR method was developed for quantification of ALV-J and compared with available quantification methods. Cell lysates with different TCID(50)s determined by cell culture and antigen capture ELISA (ag-ELISA) were used for one-tube real time RT PCR using fluorogenic probe and quantitative competitive RT-PCR (QC-RT-PCR). The results of QC-RT-PCR and real time RT-PCR were highly correlated to the TCID(50)s determined by conventional culture methods. They were also very specific, sensitive, easy to perform, reproducible, and rapid compared with conventional methods. These RT-PCR based quantification methods of ALV-J viral RNA will be useful for virological and pathogenesis studies. PMID- 11879688 TI - Use of molecular assay in diagnosis of hand, foot and mouth disease caused by enterovirus 71 or coxsackievirus A 16. AB - Hand, foot and mouth disease is a common illness in children and is usually caused by coxsackievirus A 16 and enterovirus 71. It has been noted that enterovirus 71 infection is more severe with significantly greater frequency of serious complications and fatality than coxsackievirus A 16. Therefore, it is important to develop a rapid and specific assay for discriminating coxsackievirus A 16 and enterovirus 71 in hand, foot and mouth disease outbreaks. In this study we designed two sets of RT-PCR primers specific for coxsackievirus A 16 and enterovirus 71. One hundred and eighty-nine viruses were evaluated for this molecular diagnosis assay. Among 110 enterovirus 71 strains, the enterovirus 71 specific primers gave clear signal for 107 clinical enterovirus 71 isolates and three reference enterovirus 71 strains. None of coxsackievirus A 16, other enteroviruses or non-enteroviruses show signal for enterovirus 71-specific primers. On the other hand, among 28 coxsackievirus A 16 strains, the coxsackievirus A 16-specific primers detect 27 clinical isolates and one reference strain but show no cross-reaction with other viruses. The molecular assay developed in this study provides a sensitive and specific way to distinguish coxsackievirus A 16 and enterovirus 71 induced hand, foot and mouth disease, which will be a useful rapid diagnostic method in future outbreaks. PMID- 11879689 TI - Stabilizing cold-adapted influenza virus vaccine under various storage conditions. AB - Various diluents, stabilizers, buffers, and storage conditions were assessed for their efficacy in stabilizing cold-adapted influenza virus vaccine. Frozen liquid vaccine formulations, comprised of a normal uninfected allantoic fluid diluent and an SPG (sucrose-phosphate-glutamate) stabilizer, generated complete stability of H1N1, H3N2, and Type B strains for at least 1 year of storage at -20 degrees C. The ability to store live influenza virus frozen liquid vaccines, at the moderate temperature of -20 degrees C, has not been demonstrated previously. This significant advance could facilitate influenza vaccine storage and administration in the clinic, and subsequently increase marketability. The stability of lyophilized formulations was also augmented by the addition of 2% Casitone and the control of pH with 0.066 M phosphate in the SPG stabilizer. This alternative formulation may be useful in markets where freezing is not feasible or short-term room temperature storage is necessary. PMID- 11879690 TI - Mass spectrometric identification and characterisation of the nucleocapsid protein of Menangle virus. AB - The recent emergence of novel viruses requires reliable methodology for their identification and confirmation both on a cellular and molecular level. Mass spectrometry offers a suitable approach for the identification and characterisation of viral proteins and its application is demonstrated in this study. Menangle virus is a previously unclassified member of the family Paramyxoviridae isolated in Australia in 1997. Menangle virus caused disease in pregnant pigs, and like the other newly emergent Hendra, Nipah and Tioman viruses, appears to be a virus of fruit bats (flying foxes) in the genus Pteropus. The 61 kDa gel-purified protein isolated from cell-associated Menangle virus ribonucleoprotein (RNP) was identified as the nucleocapsid protein (NP) by peptide mapping, mass spectrometry and amino acid sequencing. Over 69% of the amino acid sequence was obtained and found to be identical with that derived from gene analysis (Virology, 283 (2001), 358). The first residue of the mature NP was found to be serine (second residue in the gene derived amino acid sequence). The NP was found to be acetylated at the N-terminus (at Ser-2) and appears to be not modified by phosphorylation. PMID- 11879691 TI - Quantitation of HTLV-I proviral load by a TaqMan real-time PCR assay. AB - A quantitative real-time PCR assay was developed to measure the proviral load of human T-lymphotropic virus type I (HTLV-I) in peripheral blood mononuclear cells (PBMCs). The HTLV-I copy number was referred to the actual amount of cellular DNA by means of the quantitation of the albumin gene. Ten copies of HTLV-I DNA could be detected with 100% sensitivity, and the assay had a wide range of at least 5 log(10). Intra- and inter-assay reproducibility was evaluated using independent extractions of PBMCs from an HTLV-I-infected patient (coefficients of variation, 24 and 7% respectively). The performance of this TaqMan PCR assay, coupled with its high throughput, thus allows reliable routine follow-up of HTLV-I proviral load in infected patients. Preliminary results using clinical samples indicate a higher proviral load in patients with HTLV-I-associated myelopathy/tropical spastic paraparesis than in asymptomatic carriers, and also suggest the usefulness of this quantitative measurement to assess the etiological link between HTLV-I and adult T-cell leukaemia/lymphoma-like syndromes. PMID- 11879692 TI - Detection and subtyping of swine influenza H1N1, H1N2 and H3N2 viruses in clinical samples using two multiplex RT-PCR assays. AB - A total of 360 type A swine influenza virus-positive samples including cell culture isolates, nasal swabs or lung tissues along with 30 virus-negative samples were tested for the detection and subtyping of H1N1, H1N2 or H3N2 by two multiplex reverse transcription (RT)-PCR assays. The positive samples had been collected between 1999 and 2001 from pigs with respiratory diseases, and type A influenza virus was isolated and subtyped by hemagglutination inhibition (HI) test at the Minnesota Veterinary Diagnostic Laboratory (MVDL). Two multiplex RT PCR assays specific for H1 and H3, and N1 and N2 were developed. RT-PCR products with unique sizes characteristic of each subtype of influenza A virus were sequenced, and the sequences were demonstrated to be specific for H1N1, H1N2 or H3N2. Genomic RNAs or DNAs from 12 common swine pathogens other than type A influenza viruses were not amplified when the PCR assays were performed with these primer sets. Positive amplification reaction could be visualized with RNA extracted from up to 10(-5) dilution of each reference virus with original infectivity titer of 10(5) TCID(50)/ml. Of the 360 samples tested, swine influenza virus H1N1, H1N2 and H3N2 were identified in 200, 13 and 139 samples, respectively. The remaining eight samples were positive for both H1N1 and H3N2 viruses. The results of multiplex RT-PCR were 100% in agreement with those of virus isolation. These results demonstrate the usefulness of multiplex RT-PCR for detection and identification of influenza A virus subtypes. The results also indicate an increased occurrence of H1N2 in US swine population. PMID- 11879693 TI - Evaluation of dengue IgM detection tests using sera from patients with autoimmune diseases. AB - Three commercial dengue IgM test kits and 'in-house' IgM-capture enzyme-linked immunosorbent assay (ELISA) were examined for false positive reactions, using 49 serum samples from patients with autoimmune diseases. All the samples were found to be negative by the 'in-house' IgM-capture ELISA. Five samples were determined to be positive by the immunochromatographic test and three of the five samples were also found positive by one commercial IgM-capture ELISA kit. These results suggest that a possibility of false positive reaction should be considered when serum samples from autoimmune disease patients are tested for dengue IgM by some commercial dengue IgM test kits. PMID- 11879694 TI - Prevalence of canine coronavirus antibodies by an enzyme-linked immunosorbent assay in dogs in the south of Italy. AB - An enzyme-linked immunosorbent assay (Elisa), using as antigen canine coronavirus infected CrFK cell supernatant, was developed to detect antibodies against canine coronavirus (CCoV). Out of a total of 109 dog serum samples, 80 which were positive by routine virus neutralisation test were also Elisa positive. Seventeen samples which were negative by the virus neutralisation test, were positive by Elisa and by the confirmatory Western blotting test. The Elisa was substantially more sensitive than the virus neutralisation test in detecting antibodies to CCoV and may be used as an alternative technique to virus neutralisation. PMID- 11879695 TI - Evaluation of a Japanese quail fibrosarcoma cell line (QT-35) for use in the propagation and detection of metapneumovirus. AB - A Japanese quail fibrosarcoma cell line (QT-35) was evaluated and compared to Vero cells for its utility in metapneumovirus propagation, titration and serological detection by indirect immunofluorescence staining. Cell characteristics such as growth kinetics at different passage levels and seeding density in 96-well plates using various media formulations were studied in order to determine suitable assay parameters. Specifically, QT-35 cells supported the replication of a subgroup A metapneumovirus, strain 14/1, when maintained in DMEM containing a high level of glucose (4500 mg/l) and 2% gamma-irradiated fetal bovine serum (gamma-FBS). There appeared to be a decreased ability of metapneumovirus produced in chicken embryo fibroblast (CEF) cells to replicate to high titers in QT-35 cells, however, this apparent restriction was overcome after the second passage resulting in high titered stock. Metapneumovirus produced in Vero cells and propagated in QT-35 cells produced high titered stock after the first passage. Viral titers determined in Vero and QT-35 cells were comparable, when the latter cell line was used at passage levels < or = 20 and seeded between 5.0 x 10(4) and 1.0 x 10(5) cells/0.33 cm(2) in hgDMEM containing 10% gamma-FBS, with a reduction to 2% gamma-FBS when the virus was applied to the cell monolayers 24 h post-seeding. After infection with metapneumovirus, QT-35 cells exhibited syncytia, similar to those in metapneumovirus-infected Vero cells, which were readily detected by indirect immunofluorescent (IF) staining. PMID- 11879698 TI - The detection of tuber necrotic isolates of Potato virus Y, and the accurate discrimination of PVY(O), PVY(N) and PVY(C) strains using RT-PCR. AB - Potato tuber necrotic ringspot disease (PTNRD) is a damaging disease of potatoes, causing unsightly necrotic rings on the surface of tubers. The causal agent is thought to be tuber necrotic isolates of Potato virus Y, known as PVY(NTN). The disease spoils tubers for processing and table use, and the lack of a diagnostic method makes control especially difficult. The development of an RT-PCR assay for the reliable detection of PVY(NTN) and discrimination of all the main strains of PVY (PVY(O), PVY(N) and PVY(C)) is described. An assay was developed, exploiting a recombination site in the coat protein of PVY(NTN), allowing more reliable diagnosis of these isolates. Although the conserved nucleotide differences observed between the strains was very small, competitive PCR and mutagenically separated PCR were both employed in the development of a robust assay. The assay was found to be more reliable than the most commonly used RT-PCR method, and should prove to be an important tool in the confirmation of symptoms and for the detection of PVY(NTN) in symptomless tissue, in disease surveys and seed health schemes. PMID- 11879696 TI - Counting cytopathogenic virus adsorbed to cellulose nitrate membrane filters as a simple method for counting viruses in raw sewage and sewage effluents. AB - Using the VIRMETADEN (acronym derived from virus adsorption enumeration) method to count cytopathogenic viruses adsorbed to cellulose nitrate membrane filters from prefiltered and decontaminated sewage samples was shown to be feasible. The numbers of naturally occurring enteroviruses recovered by the VIRMETADEN method were significantly higher than those obtained by standard plaque assay. For prefiltration of sewage samples, low protein binding polyvinylidene fluoride (PVDF) membranes of 0.65 and 5 microm pore size proved to be at least as good as prefilters used previously, both in the volume that could be filtered and in the low virus retention. Regarding decontamination, 0.22 microm pore size PVDF membranes were as good as chloroform treatment. Alternatively, decontamination with 0.05% phenol followed by filtration through 0.65 microm PVDF membrane filters proved to be an excellent method for clarifying and decontaminating the sample prior to enumerating viruses using the VIRMETADEN method. It allows good recovery and greater decontaminated sample volumes. The application of VIRMETADEN to decontaminated raw sewage and secondary effluent gave recoveries of approximately 100% of vaccinal polioviruses seeded in sewage. The advantage of this approach is that the same method is applicable to sewage affluents and effluents with a simple and fast method, which implies lower material and labour costs. PMID- 11879697 TI - Temporal pattern of herpes simplex virus type 1 infection and cell death in the mouse brain stem: influence of guanosine nucleoside analogues. AB - Levels of bystander death occurring in herpes simplex virus type 1 (HSV-1) infected mouse brain stems were studied, as well as the extent to which bystander death is influenced by guanosine nucleoside analogue treatment. Consecutive sections from brain stems of HSV-1-infected mice were stained alternately for (i) viral infection and (ii) cell death (TUNEL assay). Virus antigen was detectable in brain stems on day 3 of infection, while TUNEL staining was comparatively lower. An increase in the extent of TUNEL staining was observed on day 4 of infection. Despite this increase, however, the ratio of TUNEL-stained to infection marker-stained tissue still indicated that the amount of TUNEL staining remained lower than infection staining at this time point. On days 5 and 6 of infection, TUNEL staining continued to increase and the TUNEL/infection marker ratio switched on day 6 in favour of excess TUNEL staining, which was observed in and around the foci of infection, suggesting bystander death. The excess TUNEL staining on day 6 of infection was further increased on treatment with antivirals. The significance and implications of these results are discussed with respect to the nature and mechanism of action of the TUNEL assay, dynamics of primary HSV-1 infection, immunological influences and potential effects of antiviral treatment. The potential problems of the TUNEL assay are considered in the context of viral infection and the TUNEL assay, in combination with infection marker staining, may potentially provide a model system for quantitative analysis of true bystander death during HSV infection in vivo. PMID- 11879699 TI - Optimizing dengue diagnosis by RT-PCR in IgM-positive samples: comparison of whole blood, buffy-coat and serum as clinical samples. AB - Dengue, a major public health problem in tropical and sub-tropical regions, is the most important arboviral disease of humans. Early diagnosis is very important on follow-up of infected patients, especially those at risk of the severe manifestations of this disease. Aiming at the improvement of the molecular diagnosis of these infections and due to the lack of studies that indicated the best sample for dengue virus detection by RT-PCR, viral detection by RT-PCR in blood, serum and buffy-coat of 75 IgM-positive serum samples for dengue was evaluated. Out of the 75 samples, 17 were positive for dengue using RT-PCR and from these samples, three were positive in the blood, 14 positive in the serum and eight positive in the buffy-coat. These results indicate that serum is the best clinical sample for RT-PCR amplification of dengue genomes. PMID- 11879700 TI - Real-time RT-PCR for quantitation of hepatitis C virus RNA. AB - A newly developed real-time RT-polymerase chain reaction assay for quantitation of hepatitis C virus (HCV) RNA in human plasma and serum was applied. A pair of primers and a probe (molecular beacon) were designed that are specific for the recognition of a highly conservative 5'-non-coding region (5'-NCR) in HCV genome. HCV real-time RT-PCR assay had a sensitivity of 1000 RNA copies per reaction, with a dynamic range of detection between 10(3) and 10(7) RNA copies. The coefficient variation of threshold cycle (Ct) values in intra- and inter-runs were less than 1.37 and 4.66%, respectively. The real-time RT-PCR assay on the HCV sero-positive samples yielded reproducible data, with less than 2.09% of the inter-assay variation. In order to determine its potential for clinical diagnosis, real-time RT-PCR was used to examine the HCV RNA levels in plasma from sero-positive and negative subjects, showing that the assay is highly sensitive and has specificity of 100%. It was demonstrated that the real-time RT-PCR was able to amplify HCV RNA in reference sera with seven genotypes (1A, 1B, 2B, 3A, 4, 5A and 6A) that include six major HCV genotypes circulated in the world. Since HCV is a major pathogen of post-transfusion and community-transmitted non-A, non B hepatitis, this assay has a broad application for basic and clinical investigations. PMID- 11879701 TI - Development of an ELISA for detection of antibodies to avian reovirus in chickens. AB - An enzyme-linked immunosorbent assay (ELISA) using the expressed sigmaC and sigmaB proteins which induce neutralizing antibodies as the coating antigen (sigmaC-sigmaB-ELISA) for the detection of antibodies to avian reovirus in chickens was developed and compared with serum neutralization and conventional ELISA tests. These assays were used to examine the sera from chickens vaccinated experimentally and farm chickens. The correlation rate between serum neutralization and a sigmaC-sigmaB-ELISA was 100% (156/156), and that between serum neutralization and conventional ELISA was 89.1% (139/156). The results revealed that preparation of an ELISA by using sigmaC and sigmaB of ARV as the coating antigen in detecting the field chicken sera in comparison with the conventional ELISA gave a titer more correlated to the serum neutralization test. The sigmaC-sigmaB-ELISA showed a higher correlation with the serum neutralization positive and -negative sera than that obtained with conventional ELISA. This combination antigen may thus be the best suited for preparing an ELISA for improving the determination of the immune status of chicken flocks or for detection of chicken infections with avian reovirus. PMID- 11879703 TI - Specific RT-PCR procedure for the detection of human parechovirus type 1 genome in clinical samples. AB - Among the new genera of Parechoviruses, human parechovirus type 1, formerly ECHO virus 22, has recently been recognized on the basis of distinctive biological and molecular properties. This human pathogen generally causes mild gastro-enteritis, respiratory infections and is also responsible for central nervous system infections. To ensure reliable detection, these latter infections are diagnosed by using reverse transcription polymerase chain reaction (RT-PCR) procedures. Due to genetic differences, human parechovirus type 1 genome is not detected with an enterovirus specific RT-PCR procedure. Hence, a rapid, specific and sensitive RT PCR assay was developed that can be used for the detection of human parechovirus type 1 in clinical samples. PMID- 11879702 TI - Sensitive detection of genetic variants of HIV-1 and HCV with an HIV-1/HCV assay based on transcription-mediated amplification. AB - This paper describes a comprehensive study of hepatitis C virus (HCV) and human immunodeficiency virus type 1 (HIV-1) genotype sensitivity of the transcription mediated amplification (TMA)-based HIV-1/HCV assay, developed and manufactured by Gen-Probe Incorporated (San Diego, CA) for screening human plasma specimens in blood bank settings. The TMA HIV-1/HCV assay is a qualitative, in vitro nucleic acid testing system used for initial screening. HIV-1 and HCV discriminatory assays are used to distinguish between HIV-1 and HCV infection or co-infection. In this study, multiple unique specimens representing HCV genotypes 1-6 were tested at various dilutions. The results show that the TMA HIV-1/HCV assay and the TMA HCV discriminatory assay have similar HCV genotype sensitivity, as both assays detected all six genotypes at 100 copies/ml and nearly all replicates tested at 30 copies/ml. Similarly, numerous unique specimens representing HIV-1 group M subtypes (A-G), HIV-1 group N, and group O specimens were tested at various dilutions. The TMA HIV-1/HCV assay and the TMA HIV-1 discriminatory assay were found to have similar HIV-1 subtype sensitivity; all variants at 100 copies/ml and nearly all at 30 copies/ml were detected. These results indicate that the TMA assays meet the sensitivity requirements for blood screening in blood banks worldwide. PMID- 11879704 TI - Biolistic-mediated inoculation of immature wheat embryos with Barley yellow dwarf virus-PAV. AB - Successful mechanical inoculation of plant with viruses requires an efficient method to introduce the viral pathogen into the appropriate cells of the plant. Barley yellow dwarf virus-PAV (BYDV-PAV, Luteovirus), transmitted naturally by aphids, must be inoculated into the phloem tissue to infect systemically inoculated hosts. The particle bombardment method used widely for nucleic acid transfer into plant tissues was adapted to inoculate immature embryos of winter and spring wheat cultivars with either BYDV-PAV particles or viral full-length RNAs. DAS-ELISA and RT-PCR were carried out on extracts of developed leaves at 7 weeks post-bombardment and revealed that up to 14% of bombarded embryos produced BYDV-infected wheat plants. This is the first report of an aphid-free inoculation method for BYDV. PMID- 11879705 TI - Evaluation of poliovirus vaccines for pestivirus contamination: non-specific amplification of poliovirus sequences by pan-pestivirus primers. AB - Two lots of polyvalent live vaccines for human use against poliovirus were tested by reverse transcriptase (RT) and nested PCR for the presence of contaminating pestivirus RNA. By RT-PCR, samples from both lots showed a band of approximately 450 bp instead of 300 bp for the reference pestivirus strain used as positive control. After nested PCR, the template DNA (450 bp product) was not amplified, suggesting non-specificity of the previous amplification. Sequencing analysis confirmed the non-specificity of the 450 bp bands and revealed, respectively, 80 and 77% homology with a region in the VP1 gene of poliovirus type 1 in samples 1 and 2. This suggests that more caution should be taken in interpreting the results obtained by PCR, and that they should be confirmed by nested PCR or sequencing. PMID- 11879706 TI - Erratum to "characterization and diagnostic potential of hepatitis B virus nucleocapsid expressed in E. coli and P. pastoris". AB - The hepatitis B core antigen (HBcAg) was expressed in Escherichia coli and in Pichia pastoris. A hexa-histidine tag was introduced at the C terminus of the E. coli expressed protein allowing its purification by Ni2+-chelate affinity chromatography. The P. pastoris expressed HBcAg was isolated following heat treatment. The two recombinant HBcAgs were purified further on a sucrose gradient. Mass spectrometry analysis suggested that HBcAg was N-acetylated only in P. pastoris and reaction with Ellman's reagent allowed the measurement, respectively, of 0.37 and 0.23 mole of free sulfydryl groups per mole of HBcAg monomer expressed in E. coli or P. pastoris. Electron microscopy indicated that the E. coli and the P. pastoris proteins formed capsid-like particles with, respectively, a diameter of 34 and 28 nm. Nucleic acid components were found entrapped in both particles but protected from enzymatic treatment only in the P. pastoris derived particles suggesting structural discrepancies between the two recombinant molecules. The high purity of these recombinant antigens allowed the development of a sandwich immunoassay to detect antibodies to HBcAg (anti-HBc) in human serum. The preliminary results indicate that the P. pastoris HBcAg produced intracellularly is more suitable than the renatured E. coli HBcAg for detection of anti-HBc in this diagnostic assay. PMID- 11879707 TI - Effectiveness factor in biological external convection: study in high viscosity systems. AB - A study regarding the influence of mixing on the efficiency of alginate and xanthan synthesis was carried out. The effectiveness of these two systems in terms of a dimensionless reaction convection number (N(RC)), which is a function of the power input per unit volume, was analysed and compared with low viscosity systems. The results revealed that a decrease in the power input caused a reduction in the growth rate as well as in production rate. It was observed that the effectiveness factor (eta) both for alginate production and xanthan synthesis resulted weakly dependent on the biomass content. A good correlation between effectiveness and N(RC) was obtained for the two tested models. N(RC) number could be appropriately employed in correlating the effectiveness factor for processes with viscosities from 0.001 to more than 1 kg m(-1) s(-1). Due to the parallelism with the conventional internal diffusion approach (Thiele modulus), the advantages for applying N(RC), in particular to correlate the efficiency in systems limited both for external convection and internal diffusional resistance, are shown. PMID- 11879708 TI - Effect of carbon source and aeration rate on broth rheology and fungal morphology during red pigment production by Paecilomyces sinclairii in a batch bioreactor. AB - The influence of carbon source and aeration rate on fermentation broth rheology, mycelial morphology and red pigment production of Paecilomyces sinclairii was investigated in a 5-l stirred-tank bioreactor. The characteristics of P. sinclairii grown on starch and on sucrose medium were comparatively studied: the specific growth rate in sucrose medium (0.04 h(-1)) was higher than that in starch medium, whereas the specific production rate of red pigments (0.04 gg( 1)d(-1)) was favorable in starch medium. P. sinclairii grown in sucrose medium were highly branched and showed longer hyphal lengths than that in starch medium. The consistency index (K) in sucrose medium was markedly higher than that in starch medium due to higher cell mass, while the higher values of flow behavior index (n) were indicated at the late stationary phase in starch medium. The aeration rate was varied within the ranges from 0.5 to 3.5 vvm while running the fermentation at mild agitation of 150 rpm using sucrose as the carbon source. The maximum biomass concentration of P. sinclairii was about 33 gl(-1) with an aeration rate of 1.5 vvm, whereas the maximum yield of red pigment production (4.73 gl(-1)) was achieved with 3.5 vvm. The highly branched cell morphology appeared at 1.5 vvm and the highly vacuolated cell morphology was observed in a high aeration rate (3.5 vvm). There was no significant variance in rheological parameters (K and n) between culture broths from different aeration conditions. PMID- 11879709 TI - Strategy to sequence the genome of Corynebacterium glutamicum ATCC 13032: use of a cosmid and a bacterial artificial chromosome library. AB - The initial strategy of the Corynebacterium glutamicum genome project was to sequence overlapping inserts of an ordered cosmid library. High-density colony grids of approximately 28 genome equivalents were used for the identification of overlapping clones by Southern hybridization. Altogether 18 contiguous genomic segments comprising 95 overlapping cosmids were assembled. Systematic shotgun sequencing of the assembled cosmid set revealed that only 2.84 Mb (86.6%) of the C. glutamicum genome were represented by the cosmid library. To obtain a complete genome coverage, a bacterial artificial chromosome (BAC) library of the C. glutamicum chromosome was constructed in pBeloBAC11 and used for genome mapping. The BAC library consists of 3168 BACs and represents a theoretical 63-fold coverage of the C. glutamicum genome (3.28 Mb). Southern screening of 2304 BAC clones with PCR-amplified chromosomal markers and subsequent insert terminal sequencing allowed the identification of 119 BACs covering the entire chromosome of C. glutamicum. The minimal set representing a 100% genome coverage contains 44 unique BAC clones with an average overlap of 22 kb. A total of 21 BACs represented linking clones between previously sequenced cosmid contigs and provided a valuable tool for completing the genome sequence of C. glutamicum. PMID- 11879710 TI - Measurement of hydrophobic interactions of mammalian cells grown in culture. AB - An assay was developed to measure the hydrophobic interactions of commonly used mammalian cell lines grown in culture. The assay depends on the loss of cells from an aqueous suspension following vortexing with a hydrophobic oil phase. This allowed the determination of a hydrophobicity index, which was significantly higher for Chinese Hamster Ovary (CHO) cells than either a murine hybridoma (CC9C10) or a myeloma (SP2/0). This suggests that CHO cells may have a higher intrinsic cell surface hydrophobicity. The assay was also used to study the effect of different additives on the hydrophobic interactions of the cells. A dose-dependent effect was shown for the non-ionic surfactant, Pluronic F68, in reducing the hydrophobic interaction of the CHO cells. However, the pattern of the decrease due to Pluronic F68 was different for each cell line. A higher concentration of Pluronic F68 (0.2%) was required to eliminate the hydrophobic interactions of CHO cells compared to either myelomas or hybridomas, where only 0.05% was required to reduce these interactions to a similar level. Several oils were found suitable for this assay although canola oil maximized the sensitivity of the measured changes. The assay may be useful in monitoring changes in the hydrophobic interactions of mammalian cells during growth in bioreactors. This may be important in optimizing the concentration of cell protectants such as Pluronic F68 in agitated cultures. PMID- 11879711 TI - Carbon-14 biolabelling of wine polyphenols in Vitis vinifera cell suspension cultures. AB - 14C-L-phenylalanine is incorporated into a range of polyphenolic compounds when fed to grape cell cultures. Optimisation of several parameters such as the quantity of precursor applied and the duration of metabolism led to incorporation yields of 15% and to specific activities of 875 mu Ci g(-1) in stilbenes. Purification of the products by several chromatographic steps is reported. Both trans- and cis-resveratrols were easily obtained by enzymatic hydrolysis of their corresponding glucosides, with specific activity of 1200-1400 mu Ci g(-1). The specific radioactivity obtained for all the compounds is suitable for in vivo feeding trials to trace their metabolic fate when consumed by animals and for in vitro activity mechanism studies. Indeed, these polyphenols seem to be implicated in the health benefits associated with regular and moderate wine consumption but little is known about their pharmacokinetics and cellular uptake. PMID- 11879712 TI - Modified blasticidin S resistance gene (bsrm) as a selectable marker for construction of retroviral vectors. AB - Retroviral vectors are commonly used in ex vivo gene therapy protocols. The structure of vectors basically consists of one gene of interest and a selectable marker gene. Fast selection without damaging cells is a critical step for ex vivo gene therapy protocols. Blasticidin S deaminase isolated from Bacillus cereus has a neutralizing action on the highly toxic antibiotic blasticidin S (BS). A commercially available gene coding for blasticidin S deaminase (bsr) when used to construct retroviral vectors, LBSN and LNSB, provided very low levels of BS deaminase activity, precluding their routine use in gene transfer experiments. However, with the introduction of specific mutations into the bsr gene based on the Kozak consensus sequences and deletion of a 5' untranslated sequence to generate bsrm, we were able to construct a retroviral vector encoding resistance to high doses of BS (at least 16-fold above the usual lethal dose in NIH3T3 cells), showing that bsrm/BS may provide a useful system for selection of transduced mammalian cells. PMID- 11879713 TI - Instantaneous evaluation of mammalian cell culture growth rates through analysis of the mitotic index. AB - Since a culture increases in cell number when dividing cells separate into two newborn cells, the fraction of mitotic cells in a growing cell population directly reflects the overall growth behavior of a cell culture. To rapidly assess the effects of growth conditions on the fraction of mitotic cells we have employed an antibody specific for the phosphorylated form of histone H3 for the identification of mitotic cells using flow cytometry. The phosphorylation of histone H3 closely correlates with the chromosomal condensation that accompanies the onset of mitosis, and, therefore, it represents a convenient marker for dividing cells. We have optimized the protocol for the staining of mitotic cells for both Chinese hamster ovary and hybridoma cell cultures. Fluorescence micrographs taken of stained cells show that cells in the various stages of mitosis can be detected based on the morphological characteristics of the chromosomes. The variation in the mitotic cell fraction has been determined throughout the batch growth phases of cultures under different growth conditions. The dynamics of the mitotic index show that balanced growth was never truly reached and that the growth rate is in fact quite variable for these cultures since large variations in the mitotic index are observed. In addition, a large increase in the fraction of mitotic cells just prior to the exponential growth phase for all cultures indicates that they are partially synchronized at the exit from the lag phase. According to a two-staged, age structured population balance model, the mitotic index is directly proportional to the growth rate of a culture. The proportionality constant for this case is shown to be the time required for cells to progress through mitosis. This time is believed to be constant for a particular cell line, as shown by experimental data. Thus, growth rates can be determined solely by measurement of the fraction of cells in mitosis. The mitotic index measurements were then used to calculate the growth in cell number of the cultures, and these simulations accurately reflect observed cell counts. Other simulations also show that changes in cell growth can be predicted before they are reflected in the cell count data. This technique can be used as a sensitive indicator of cell growth and could be useful as a process monitoring technique and for developing better feeding strategies for animal cell cultures. PMID- 11879714 TI - Non-destructive monitoring of rpoS promoter activity as stress marker for evaluating cellular physiological status. AB - To monitor the extent of cellular physiological stress, the activity of the rpoS promoter was evaluated as a marker of the stress pathway. A reporter plasmid was constructed by inserting the GFPuv gene under the rpoS promoter and used to transform Escherichia coli cells. The fluorescence of the GFPuv protein was measured in intact cells in a non-destructive manner. The physiological status of the cells could be conveniently monitored using the rpoS-GFPuv reporter gene with respect to the cellular growth phase and to elevated ethanol and NaCl concentrations as two examples of environmental stress factors. Comparison of the response of different E. coli strains demonstrated an essential role of the relA gene in the induction of the rpoS-GFPuv reporter gene. PMID- 11879715 TI - Simvastatin retards progression of retinopathy in diabetic patients with hypercholesterolemia. AB - Besides hyperglycemia and hypertension, a recently recognized risk factor for diabetic retinopathy (DR) appears to be hyperlipidemia. While studies using earlier generation lipid lowering agents in DR were disappointing, a randomized trial using HMG-CoA Reductase Inhibitors has strong rationale, though hitherto not attempted. The aim of the present study was to compare the HMG-CoA Reductase Inhibitor, simvastatin, with placebo in patients having DR in a double-blind randomized placebo-controlled trial. Fifty patients with diabetes mellitus (Type 1 and 2) with good glycemic control and hypercholesterolemia and having DR (non clinically significant macular edema and visual acuity 6/24 or better) in either or both eyes were randomized to simvastatin 20-mg per day or placebo, and were followed up for 180 days. On simvastatin therapy, total cholesterol and low density lipoprotein cholesterol (LDL-C) decreased (P < 0.001, respectively), and the level of high-density lipoprotein cholesterol (HDL-C) increased (P < 0.001). VA improved in four patients using simvastatin, (not statistically different from placebo group) and worsening of VA occurred in seven patients in the placebo group and none in the simvastatin group (P = 0.009). Fundus fluorescein angiography and color fundus photograph showed improvement in one patient in the simvastatin group, while seven patients showed worsening in the placebo group (P = 0.009). The observations of the current study suggest that the HMG-CoA Reductase Inhibitor simvastatin significantly retards the progression of retinopathy in diabetic patients with hypercholesterolemia. The potential of this class of drugs for the primary prevention of DR and other microvascular complications needs to be explored further. PMID- 11879716 TI - Sudden onset of diabetes with ketoacidosis in a patient treated with FK506/tacrolimus. AB - We report a 43-year-old man who presented diabetic ketoacidosis 1 year after receiving kidney transplantation. He was a recipient of renal transplantation treated with metyl-prednisolone and tacrolimus regimen. The serum level of tacrolimus was 12.4 ng/ml, and he showed hyperphagia before a month of admission. A week before admission, he was aware of polydipsia, polyuria, and general fatigue. He visited our hospital and was found to have severe hyperglycemia (925 mg/dl), significant ketosis and mild metabolic acidosis (pH 7.341), although he had not been diagnosed as diabetes mellitus. He administrated in our hospital, and was treated with insulin for 5 weeks. He was not obese (BMI = 18.2 kg/m(2)) and had no family history of type 2 diabetes. He was finally treated with diet therapy alone. The 24 h urine C-peptide secretion on the third hospital day was low (8.4 microg per day). However, no autoantibodies against pancreatic islets were positive, and his insulin secretion was recovered at discharge suggesting that he was not type 1 diabetes. Although, tacrolimus has been reported to cause or worsen diabetes mellitus, the present case suggests that it could cause severe decrease in insulin secretion which leading to diabetic ketoacidosis in lean subject without previous history of diabetes mellitus. PMID- 11879717 TI - Increase of serum phosphatidylcholine hydroperoxide dependent on glycemic control in type 2 diabetic patients. AB - In order to clarify the relationship between serum phosphatidylcholine hydroperoxide (PCOOH) levels and blood glucose control in type 2 diabetes patients (DM), DM (n = 61) and normal control (n = 11) were enrolled. High density lipoprotein (HDL) was separated from serum by the addition of sodium phosphotungstate and magnesium chloride, and the precipitated fraction was prepared as non-HDL. Phospholipids were extracted from whole serum, non-HDL and HDL to estimate PCOOH level with chemiluminescence high performance liquid chromatography (CL-HPLC). PCOOH level (nmol/l, mean +/- S.D.) was higher in DM than in control (33.1 +/- 9.5 vs. 23.0 +/- 8.2 for serum; P < 0.01, 17.0 +/- 5.5 vs. 10.6 +/- 3.8 for non-HDL; P < 0.01, and 16.1 +/- 6.3 vs. 12.3 +/- 5.5 for HDL; not significant, respectively). DM was divided into five groups according to hemoglobin A(1c) (HbA(1c)) levels (%): (1) less than 6, (2) 6-6.4, (3) 6.5-6.9 (4) 7.0-7.4, and (5) over than 7.5. Increase of PCOOH levels was dependent on HbA(1c). We concluded that (1) serum and non-HDL PCOOH increased in DM, (2) the level was strongly correlated with diabetic control, and (3) approximately a half amount of serum PCOOH was present in HDL of both control and DM. PMID- 11879718 TI - Care resource utilization and direct costs incurred by people with diabetes in a Spanish hospital. AB - The objective of our study was to determine the hospital care resource utilization and direct medical costs incurred for in-patients with diabetes compared with non-diabetic in-patients. The data were obtained from the records division of the Puerta del Mar University Hospital, an 800-bed tertiary care hospital in Cadiz in the south of Spain. We assessed the rate of hospital admissions, length of stay, readmissions, mortality and costs for both diabetic and non-diabetic people. People with diabetes accounted for 10.9% of total hospital discharges (2453 discharges), 15.3% of total stays (30,771 days) and 16.1% of total cost (Euro 7,417,688). We estimated a hospitalization rate of 135 per 1000 persons with diabetes (compared with 95 per 1000 non-diabetic persons). Diabetic patients were hospitalized, on average, for 4 days longer than non diabetic patients (12.5 +/- 14.5 (+/-SD) vs 8.5 +/- 10.6 days; P < 0.001) and had higher risks of readmission (RR: 2.29 (95% CI: 1.91-2.74)) and of mortality during the in-patient period (2.29 (1.91-2.74)) than non-diabetic patients. The overall hospitalization cost was significantly higher in diabetic (Euro 3023 +/- 3463) than in non-diabetic patients (Euro 1949 +/- 2528), 55% higher than the average general cost. We conclude that the hospital care resource utilization and economic burden due to diabetes mellitus in our hospital is substantial and disproportionate to the number of affected people. PMID- 11879719 TI - Diabetic foot disease and foot care in a Caribbean community. AB - We estimated the prevalence of foot symptoms and disease and evaluated foot care practices in a primary care based sample including 2106 people with diabetes in Trinidad. Symptoms of neuropathy were reported by 1030 (49%), previous foot ulceration by 257 (12%), and amputation by 92 (4%). Previous foot ulceration was associated with longer duration of diabetes (odds ratio 1.05, (95% CI 1.04-1.06) per year) and greater severity of neuropathy symptoms (1.17 (1.10-1.24) per unit increase in score). A history of foot ulceration gave relative odds of amputation of 16.3 (8.1-32.9). In those with previous foot ulceration, 120 (47%) went barefoot in the home, and 44 (17%) went barefoot outside the house. Overall, 1491 (71%) subjects reported they cut their toenails themselves, help was provided by a friend or relative to 584 (28%) and by a nurse or chiropodist to 13 ( < 1%). Most patients (1320, 63%) reported that they would treat a cut or blister on the foot themselves, while only 650 (31%) would attend for health care. Diabetic foot disease is common but care practices predispose to foot injury. Implementation of a strategy to improve care of the feet is needed. PMID- 11879720 TI - GAD65 and IA-2 autoantibodies are common in a subset of siblings of Sardinian Type 2 diabetes families. AB - Type 1 diabetes in Sardinia is very common in children, and we hypothesized that Latent Autoimmune Diabetes of Adult (LADA) might constitute a significant proportion of diabetes in adult Sardinian subjects. Since Type 2 diabetes is a familial disorder, we tested this hypothesis by investigating the prevalence of GAD65 and IA-2 autoantibodies (Ab) in Type 2 diabetes multiplex families of Sardinian ancestry enrolled in the Study Group for the Genetics of Diabetes in Sardinia. METHODS: A total of 684 individuals were ascertained from 252 Sardinian Type 2 diabetes multiplex families with 2.4 affected siblings per family comprising 190 families with two affected, 37 with three, 15 with four, 7 with five, and 3 with six, in addition to 80 unaffected siblings. Controls were household contacts representing 204 healthy spouses of affected siblings. Diagnosis was at 35-69 years of age and insulin was not given in the first 4 years after diagnosis. GAD65Ab and IA-2Ab were determined in standard radioligand binding assays. RESULTS: Among affected siblings GAD65Ab were positive in 8.8% of insulin-treated (n = 137; P = 0.0006), in 2.5% of non-insulin-treated (n = 467), and in 1.2% of non-diabetic siblings (n = 80) compared with 0.5% of controls (n = 204). IA-2Ab was positive in 6.6% insulin-treated (P = 0.04), 2.1% non-insulin treated, and 2.5% non-diabetic siblings compared with 1.5% of controls. CONCLUSION: A high frequency of GAD65Ab and IA-2Ab as markers of Type 1 diabetes was found among Type 2 diabetes siblings from Sardinian multiplex families despite excluding those who had been treated with insulin during the first 4 years of disease. Our data support the hypothesis that LADA may be common in Sardinian Type 2 diabetes and stress the importance of investigating markers of Type 1 diabetes in studies of Type 2 diabetes. PMID- 11879721 TI - Determinants of ambulatory care in a defined adult Swedish diabetic population. AB - AIMS: To investigate factors associated with health care utilisation in ambulatory diabetes care in relation to complications attributable to diabetes mellitus in an adult diabetic population. METHODS: A cross-sectional study; standardised interview, physical examination, and an evaluation of medical records, comprising all known diabetic subjects living in six primary health care districts in southern Sweden (N = 1861, aged > 25 years; 90.1% participation). RESULTS: People managed by specialists (17.2%) had more complications related to diabetes and were more often treated with insulin. Persons managed in health care centres with a diabetes nurse specialist used insulin more often, used self monitoring of blood glucose (SMBG) more regularly, lived on their own, and used meals on wheels and Community Care Alarm Service more frequently than those managed in other health centres. Multiple logistic regression analysis showed high utilisation of office visits to physicians ( > 5 visits) to be mainly associated with the presence of a foot ulcer (OR (95% CI) 2.1 (1.4-3.3)), congestive heart failure (1.6 (1.1-2.3)), and cardio-cerebrovascular disease (1.4 (1.1-1.9)). High utilisation of visits to other care-givers ( > 4 visits) was related to current or previous foot ulcers (2.4 (1.5-3.7) and 2.1 (1.2-3.5)), meals on wheels (1.9 (1.2-3.0)), and treatment with insulin (1.6 (1.2-2.1)). CONCLUSIONS: High utilisation of ambulatory diabetes care was mainly associated with health status and complications related to diabetes, particularly diabetic foot ulcers. Organisational factors such as managed care with access to a diabetes nurse specialist in a health care centre was related to increased use of self-monitoring of blood glucose and insulin treatment. Visits to other care givers were associated with access to social welfare. In diabetes care, activities to promote health and prevent complications need to be stressed. PMID- 11879722 TI - Association of plasma lipid levels with apolipoprotein E polymorphism in Type 2 diabetes. AB - OBJECTIVE: To evaluate the distribution of apolipoprotein E polymorphism in patients with Type 2 diabetes and their impact on plasma lipid levels. SUBJECTS: Unrelated Type 2 diabetic patients (n = 298) treated by diet and sulfonylurea and not receiving lipid-lowering regimens, elderly (n = 98) and young (n = 101)unrelated healthy control subjects in Hungary. METHODS: Apolipoprotein E genotypes were identified by PCR amplification and subsequent restriction endonuclease digestion. RESULTS: The distribution of the most frequent genotypes in the diabetes group was E2/3 8.7%, E3/3 78.2%, E3/4 12.8%, in the elderly group E2/3 9.2%, E3/3 78.6%, E3/4 12.2% and in the young group E2/3 11.9%, E3/3 62.4%, E3/4 24.8%. The frequencies of allele e4 in the diabetes and in the elderly control group were significantly lower than in the young control group (both P < 0.05). Associations were found between the e4 allele and increased triglyceride level in the diabetes group, the e2 allele and decreased total cholesterol and LDL-cholesterol levels both in the elderly and young control groups (both P < 0.01). CONCLUSION: The lower frequency of allele e4 in both the elderly and diabetes groups, may be explained by an increased morbidity and mortality in middle-aged carriers of apo e4 allele. The increased risk of e4 carriers in Type 2 diabetes may be partly mediated by a higher triglyceride level. PMID- 11879723 TI - Insulin responses to sulfonylureas. PMID- 11879725 TI - Insulin resistance and clustering of atherogenic risk factors in women belonging to low socio-economic strata in urban slums of North India. PMID- 11879726 TI - Lymphoproliferative response to glutamic acid decarboxylase in fibrocalculous pancreatopathy. PMID- 11879727 TI - Inhibition of copper amine oxidases by pyridine-derived aldoximes and ketoximes. AB - The reactions of pea diamine oxidase (PSAO) and 2-phenylethylamine oxidase from Arthrobacter globiformis (AGAO) with pyridine-derived oximes were studied. Pyridine carbaldoximes and alkyl pyridyl ketoximes act as strong non-competitive inhibitors of the enzymes. The inhibition constants K(i) of these compounds vary between 10(-4) and 10(-5) M, for AGAO and some of the studied oximes were found even micromolar K(i) values. The presence of pyridine moiety in the studied compounds has remarkable influence on the inhibition potency. Elementary oximes lacking the heterocyclic ring, i.e., aliphatic (acetone oxime), alicyclic (cyclohexanone oxime) and aromatic (benzaldoxime), are considerably weaker non competitive inhibitors (K(i) similar to 10(-3) or 10(-2) M). The position of the pyridine ring substitution by -C(R)=NOH group does not play a significant role for the inhibition potency of the studied oxime compounds. If the pyridine nitrogen is quaternised (in hydroxyiminomethyl-1-methylpyridinium iodides), the compound looses its inhibitory properties. Extended length of alkyl substituents on the ketoxime group of alkyl pyridyl ketoximes increases the K(i) value. The enzyme-bound copper represents one of possible target sites for pyridine-derived oxime inhibitors. The addition of an alkyl pyridyl ketoxime or a pyridine carbaldoxime to a native PSAO sample perturbs the absorption spectrum of the enzyme (by an absorption increase in the region 300-400 nm) that is not observed in the spectrum of reacted PSAO apoenzyme. However, an additional formation of hydrogen bonds with amino acid side-chains at the active site should be considered, namely for 3- and 4-substituted pyridine derivatives. PMID- 11879728 TI - Cloning of the PpMSH-2 cDNA of Physcomitrella patens, a moss in which gene targeting by homologous recombination occurs at high frequency. AB - In the moss Physcomitrella patens integrative transformants from homologous recombination are obtained at an efficiency comparable to that found for yeast. This property, unique in the plant kingdom, allows the knockout of specific genes. It also makes the moss a convenient model to study the regulation of homologous recombination in plants. We used degenerate oligonucleotides designed from AtMSH2 from Arabidopsis thaliana and other known MutS homologues to isolate the P. patens MSH2 (PpMSH2) cDNA. The deduced sequence of the PpMSH2 protein is respectively 60.8% and 59.6% identical to the maize and A. thaliana MSH2. Phylogenic studies show that PpMSH2 is closely related to the group of plant MSH2 proteins. Southern analysis reveals that the gene exists as a single copy in the P. patens genome. PMID- 11879729 TI - Developmentally programmed excision of internal DNA sequences in Paramecium aurelia. AB - The development of a new somatic nucleus (macronucleus) during sexual reproduction of the ciliate Paramecium aurelia involves reproducible chromosomal rearrangements that affect the entire germline genome. Macronuclear development can be induced experimentally, which makes P. aurelia an attractive model for the study of the mechanism and the regulation of DNA rearrangements. Two major types of rearrangements have been identified: the fragmentation of the germline chromosomes, followed by the formation of the new macronuclear chromosome ends in association with imprecise DNA elimination, and the precise excision of internal eliminated sequences (IESs). All IESs identified so far are short, A/T rich and non-coding elements. They are flanked by a direct repeat of a 5'-TA-3' dinucleotide, a single copy of which remains at the macronuclear junction after excision. The number of these single-copy sequences has been estimated to be around 60,000 per haploid genome. This review focuses on the current knowledge about the genetic and epigenetic determinants of IES elimination in P. aurelia, the analysis of excision products, and the tightly regulated timing of excision throughout macronuclear development. Several models for the molecular mechanism of IES excision will be discussed in relation to those proposed for DNA elimination in other ciliates. PMID- 11879730 TI - How to characterize meiotic functions in plants? AB - Our understanding of plant meiosis is rapidly increasing thanks to the model Arabidopsis thaliana. Here we present the results of a screening for meiotic mutants carried out with a library containing 30,719 T-DNA insertion lines. An average of one mutant per 1000 lines was recovered. Several phenotypic classes could be distinguished and are presented. In parallel, 39 proteins known to be involved in meiosis in non-plant organisms were chosen and a search was performed for homologue sequences in the completed Arabidopsis thaliana genome. Approximately 30% of the meiotic related sequences showed similarities with one or several Arabidopsis putative genes. The relevance of forward versus reverse genetics in order to characterize meiotic functions is discussed. PMID- 11879731 TI - Chromatin structure and dynamics: functional implications. AB - In eucaryotes, DNA packaging into nucleosomes and its organization in a chromatin fiber generate constraints for all processes involving DNA, such as DNA replication, -repair, -recombination, and -transcription. Transient changes in chromatin structure allow overcoming these constraints with different requirements in regions where processes described above are initiated. Mechanisms involved in chromatin dynamics are complex. Multiprotein complexes which can contain histone-acetyltransferase, -deacetylase, -methyltransferase or -kinase activities are targeted by regulatory factors to precise regions of the genome. These enzymes have been shown to modify histone-tails within specific nucleosomes. Post-translational modifications of histone-tails constitute a code that is thought to contribute to the nucleosome or to the chromatin fiber remodeling, either directly, or through the recruitment of other proteins. Other multiprotein complexes, such as ATP-dependent remodeling complexes, play an essential role in chromatin fiber dynamics allowing nucleosome sliding and redistribution on the DNA. We will focus here on the chromatin structure and its consequences for DNA damaging, replication, repair, and transcription and we will discuss the mechanisms of chromatin remodeling. PMID- 11879732 TI - Genetic evidence that the elevated levels of Escherichia coli helicase II antagonize recombinational DNA repair. AB - Some phages survive irradiation much better upon multiple than upon single infection, a phenomenon known as multiplicity reactivation (MR). Long ago MR of UV-irradiated lambda red phage in E. coli cells was shown to be a manifestation of recA-dependent recombinational DNA repair. We used this experimental model to assess the influence of helicase II on the type of recombinational repair responsible for MR. Since helicase II is encoded by the SOS-inducible uvrD gene, SOS-inducing treatments such as irradiating recA(+) or heating recA441 cells were used. We found: i) that MR was abolished by the SOS-inducing treatments; ii) that in uvrD background these treatments did not affect MR; and iii) that the presence of a high-copy plasmid vector carrying the uvrD(+) allele together with its natural promoter region was sufficient to block MR. From these results we infer that helicase II is able to antagonize the type of recA-dependent recombinational repair acting on multiple copies of UV-damaged lambda DNA and that its anti recombinogenic activity is operative at elevated levels only. PMID- 11879733 TI - Construction and characterization of a recombinant chimeric plasminogen activator consisting of a fibrin peptide and a low molecular mass single-chain urokinase. AB - A recombinant chimeric plasminogen activator (f beta/scuPA-32k), with a fibrin beta-chain peptide (comprising Gly15 through Arg 42) linked to the N-terminal of a low molecular mass (32 kDa) single-chain urokinase (scuPA-32k, comprising Leu144 through Leu 411) via a 50 amino acid linker sequence, was produced by expression the corresponding chimeric cDNA in Escherichia coli cells. After refolding in vitro, the chimeric protein was purified to homogeneity by zinc chelate-Sepharose chromatography, Sephacryl S200 chromatography and benzamidine Sepharose chromatography in sequence. The apparent molecular mass was 36 kDa shown by SDS-PAGE analysis. The special activity was 87,000 IU/mg detected by fibrin plate determination. F beta/scuPA-32k could directly activate plasminogen following Michaelis-Menten kinetics with K(m) = 0.52 microM and k(2) = 0.0024 s( 1). Mediated by plasmin, the single-chain molecule could be converted to the active two-chain molecule. The chimeric protein had 3.3 times higher fibrin affinity than scuPA-32k in the fibrin concentration of 3.2 mg/mL, while the chimeric protein inhibited the fibrin clotting and platelet aggregation. F beta/scuPA-32k showed a higher thrombolytic potency in vitro plasma clot lysis than scuPA-32k and depleted less fibrinogen in plasma. These results showed that the chimeric protein had not only higher fibrinolytic activity but also anti thrombus activity. Further evaluation of the thrombolytic potential in appropriate animal models is required. PMID- 11879734 TI - Inactivation of NADP(+)-dependent isocitrate dehydrogenase by reactive oxygen species. AB - Recently, we demonstrated that the control of cytosolic and mitochondrial redox balance and the cellular defense against oxidative damage is one of the primary functions of NADP(+)-dependent isocitrate dehydrogenase (ICDH) through supply of NADPH for antioxidant systems. When exposed to various reactive oxygen species such as hydrogen peroxide, singlet oxygen generated by photoactivated dye, superoxide anion, and hydroxyl radical produced by metal-catalyzed Fenton reactions, ICDH was susceptible to oxidative modification and damage, which was indicated by the loss of activity, fragmentation of the peptide as well as by the formation of carbonyl groups. Oxidative damage to ICDH was inhibited by antioxidant enzymes, free radical scavengers, and spin-trapping agents. The structural alterations of modified enzymes were indicated by the increase in thermal instability and binding of the hydrophobic probe 8-anilino-1-naphthalene sulfonic acid (ANSA). The reactive oxygen species-mediated damage to ICDH may result in the perturbation of cellular antioxidant defense mechanisms and subsequently lead to a pro-oxidant condition. PMID- 11879736 TI - Preparation and in vitro evaluation of solid dispersions of halofantrine. AB - The low aqueous solubility of halofantrine (HF) and its low bioavailability from commercially available tablets (Halfan) suggested the formulation of solid dispersions (SDs) of HF to reduce its particle size and improve its wettability and aqueous solubility. Preformulation studies involved the development of a high performance liquid chromatography (HPLC) method for the analysis of HF. In addition, solubility studies were conducted on HF in aqueous solutions containing different concentrations of various carriers. Formulation studies included the preparation of SDs and physical mixtures (PMs) of HF with different carriers and their physicochemical characterization using differential scanning calorimetry (DSC), Fourier-Transform infra-red (FT-IR) spectroscopy and dissolution studies. A 3-month stability study at elevated temperatures was conducted on representative SDs of HF with selected carriers. PMID- 11879735 TI - Colon-specific drug delivery: new approaches and in vitro/in vivo evaluation. AB - The necessity and advantages of colon-specific drug delivery systems have been well recognized and documented. In the past, the primary approaches to obtain colon-specific delivery achieved limited success and included prodrugs, pH- and time-dependent systems, and microflora-activated systems. Precise colon drug delivery requires that the triggering mechanism in the delivery system only respond to the physiological conditions particular to the colon. Hence, continuous efforts have been focused on designing colon-specific delivery systems with improved site specificity and versatile drug release kinetics to accommodate different therapeutic needs. Among the systems developed most recently for colon specific delivery, four systems were unique in terms of achieving in vivo site specificity, design rationale, and feasibility of the manufacturing process (pressure-controlled colon delivery capsules (PCDCs), CODES, colonic drug delivery system based on pectin and galactomannan coating, and Azo hydrogels). The focus of this review is to provide detailed descriptions of the four systems, in particular, and in vitro/in vivo evaluation of colon-specific drug delivery systems, in general. PMID- 11879737 TI - Dependence of low-frequency sonophoresis on ultrasound parameters; distance of the horn and intensity. AB - Sonophoresis at a frequency of 20 kHz has been shown to enhance transdermal drug delivery, a phenomenon referred to as low-frequency sonophoresis. This study provides an investigation of the dependence of low-frequency sonophoresis on various ultrasound parameters, including the distance of the horn from the skin, intensity, and frequency. We performed in vitro experiments with full thickness pig skin to measure enhancements of skin conductivity and drug permeability. Ultrasound was applied to pretreat the skin using a sonicator operating at a frequency of either 20 or 40 kHz. We also measured pitting of aluminum foil to measure cavitation, which is the principal mechanism of low-frequency sonophoresis. The skin conductivity enhancement was found to be inversely proportional to the distance of the horn from the skin. As the intensity increased, skin conductivity enhancement also increased up to a certain threshold, and then dropped off. The intensities (I(max)) at which maximum enhancement occur are about 14 W/cm2 for 20 kHz and 17 W/cm2 for 40 kHz. These findings may be useful in optimizing low-frequency sonophoresis. Overall, the dependence of transport on ultrasound parameters is similar to that of aluminum foil pitting. These results support the role of cavitation in low-frequency sonophoresis. PMID- 11879738 TI - A novel thermo-responsive drug delivery system with positive controlled release. AB - The model drug, 5-fluorouracil (5-FU) was loaded into the poly(N isopropylacrylamide) (PNIPA) hydrogel at 25 degrees C, then the drug-loaded, swollen hydrogel sample was carefully enveloped in the dialysis bag to form a novel thermo-responsive drug delivery system (DDS). The concentration of released 5-FU was monitored at 266 nm on the UV spectrophotometer. We found that this novel DDS provides a positive drug release pattern and the drug, 5-FU, was released faster at the increased temperature (37 degrees C, >25 degrees C) than the one at the decreased temperature (10 degrees C, <25 degrees C). This was attributed to the double control of the thermo-sensitivity of the hydrogel matrix and the dialysis membrane. By employing the fast response PNIPA hydrogel instead of the conventional hydrogel in this novel DDS, we can further control the drug release rate and/or drug release amount etc., without changing the positive, thermo-responsive drug release pattern. PMID- 11879739 TI - Design and evaluation of a mucoadhesive therapeutic agent delivery system for postoperative chemotherapy in superficial bladder cancer. AB - The most common treatment method is known as the transurethral resection (TUR) for the therapy of bladder cancer. Unfortunately, because of the recurrency of the tumoral tissues after TUR the chemotherapy or immunotherapy should be performed. In these kind of therapies the pharmacotherapeutics are infused intravesically into the bladder after TUR periodically (i.e. upto 6--36 weeks, each week). But these therapies are having very big problems (i.e. disturbancy to patients, adjustment of the suitable dosage, loss of active agents without using etc.). An alternative approach can be proposed as to design a pharmacotherapeutic agent delivery system, which will supply the suitable dosage of the agent for a certain time period to solve those problems. In this study; the pharmacotherapeutic agent (i.e. Mytomycin-C) delivery system was prepared by using a mucoadhesive polymer (i.e. chitosan) in the form of cylindirical geometry to facilitate the insertion of the carrier for in vivo studies. The chitosan carriers were prepared by cross-linking during the solvent evaporation technique. In the preparation of the chitosan carriers the chitosan polymers with different molecular weights, different amounts of cross-linker (i.e. glutaraldehyde) and different amounts of pharmacotherapeutic agent were used to obtain desired attachment onto the bladder wall and optimum release rate of the agent. On the other hand because of the gelous structure of the chitosan, the swelling behaviour of the polymer was evaluated by gravimetric determinations in aqueous media periodically. PMID- 11879740 TI - Development of novel topical tranexamic acid liposome formulations. AB - The aims of this study were to develop novel liposome formulations for tranexamic acid (TA) from various lipid compositions [neutral (hydrogenated soya phosphatidylcholine and cholesterol), positive (stearylamine) or negative (dicetyl phosphate) charged lipid], and to investigate the effects of concentrations of TA (5 and 10% in DI water) and charges on the physicochemical properties of liposomes. Liposomes were prepared by chloroform film method with sonication. The physical (appearance, pH, size, morphology) and chemical (drug encapsulation efficiency, transition temperature, enthalpy of transition) properties of liposomes were characterized. The TA contents were determined spectrophotometrically at 415 nm, following derivatization with 2,4,6 trinitrobenzosulfonic acid. The charged liposomes demonstrated better physical stability than the neutral liposomes. The percentages of TA entrapped in all liposome formulations varied between 13.2 and 15.6%, and were independent of TA concentrations and charges of liposomes. Charges affected the physical stability, pH and size of liposomes. The particle sizes of negative blank and positive liposomes (with and without the entrapped drug) were approximately 10 times larger than the negative liposome with the entrapped TA. The multilamellar 7:2:1 molar ratio of hydrogenated soy phosphatidylcholine/cholesterol/dicetyl phosphate entrapped with 10% TA liposome (10%TA,-) was selected for further release study, due to its high physical stability, small particle size and relatively high drug encapsulation efficiency. PMID- 11879741 TI - Permeability of human intestinal mucosa using a continuous flow-through perfusion system. AB - Continued interest in in vitro methods for performing bioavailability/bioequivalence (BA/BE) studies for drug registration purposes, prompted us to investigate the suitability of a continuous flow-through perfusion system to determine diffusion of a wide variety of permeants, through human intestinal mucosa. Permeability of fresh and frozen intestinal mucosa towards water, 17beta-estradiol, sumatriptan, arecoline and vasopressin was compared. Furthermore, diffusion studies of water, sumatriptan, arecoline, arecaidine, estradiol, cyclosporin and vasopressin across frozen/thawed intestinal mucosa specimens (-85 degrees C) were performed. No statistically significant differences between the flux values of the five compounds tested across fresh and frozen intestinal tissue, were found. Furthermore, it was demonstrated that the flux rates of the various compounds across these tissues decreased with increasing molecular size. However, the flux rates across frozen intestinal mucosa for compounds with molecular weights >300 Da, were low. Flux rates for the compounds studied across frozen/thawed human vaginal and buccal mucosa were 36 160% higher than those across frozen intestinal mucosa. We concluded that the continuous flow-through perfusion system used shows promise as an in vitro method for permeability determination through intestinal mucosa. However, other human mucosa e.g. vaginal mucosa, may have to be considered as alternatives to intestinal mucosa if therapeutic agents with molecular weights >500 Da are to be compared for in vitro BA/BE purposes, and further studies in this respect are warranted. PMID- 11879742 TI - Amylopectin as a subcoating material improves the acidic resistance of enteric coated pellets containing a freely soluble drug. AB - The effect of an aqueous amylopectin subcoating on the acidic resistance and dissolution behaviour of enteric-coated pellets was studied. Freely water-soluble riboflavin sodium phosphate (RSP) was used as a model drug, and microcrystalline cellulose (MCC) and lactose as fillers in the pellet cores. The pellets were subcoated with 5% aqueous amylopectin solution or with 5% hydroxypropyl methylcellulose (HPMC) solution, and subsequently film-coated with aqueous dispersion of cellulose acetate phthalate (CAP). Drug release of enteric-coated pellets was investigated by confocal laser scanning microscopy (CLSM). Dissolution tests showed that amylopectin subcoating improved the acidic resistance of the enteric-coated pellets in 0.1 N hydrochloric acid (HCl) compared with HPMC subcoating. As the amylopectin subcoating load was increased to 4% and the aqueous CAP coating load to 35%, the coated pellets resisted in 0.1 N HCl solution for approximately 1 h (the amount of drug released was below 10%), and they dissolved in the SIF without enzymes in less than 10 min. Confocal microscopy images and profiles of mean fluorescence intensities of RSP (obtained in the range of the interface of the pellet core and the film and the film coating surface) showed consistent results with dissolution tests. It seems that amylopectin subcoating can prevent the influx of the dissolution medium into the pellet core, and thus decrease the premature dissolution and release of the drug from the enteric-coated pellets in 0.1 N HCl solution. The drug release mechanism appeared to be osmotically driven release, and followed by diffusion through the polymer film. PMID- 11879743 TI - Stomach-specific anti-H. pylori therapy. I: Preparation and characterization of tetracyline-loaded chitosan microspheres. AB - The main objective of the study was to develop a stomach-specific drug delivery system to increase the efficacy of tetracycline against Helicobacter pylori. Chitosan microspheres were prepared by ionic cross-linking and precipitation with sodium sulfate. Two different methods were used for drug loading. In method I, tetracycline was mixed with chitosan solution before the simultaneous cross linking and precipitation. In method II, the drug was incubated with pre-formed microspheres for 48 h. The cumulative amount of tetracycline that was released from chitosan microspheres and the stability of the drug was examined in different pH medium at 37 degrees C. Microspheres with a spherical shape and an average diameter of 2.0-3.0 microm were formed. When the drug was added to the polymer solution before cross-linking and precipitation only 8% (w/w) was optimally incorporated in the final microsphere formulation. When the drug was incubated with the pre-formed microspheres, on the other hand, a maximum of 69% (w/w) could be loaded. Thirty percent of tetracycline either in solution or when released from microspheres was found to degrade at pH 1.2 in 12 h. The preliminary results from this study suggest that chitosan microspheres can be used to incorporate antibiotic drugs and may be effective when administered locally in the stomach against H. pylori. PMID- 11879744 TI - Transdermal iontophoresis of sodium nonivamide acetate. V. Combined effect of physical enhancement methods. AB - The effect of iontophoresis combined with treatment of other physical enhancement methods such as electroporation, low frequency ultrasound, and erbium:YAG (yttrium-aluminum-garnet) laser on the transdermal delivery of sodium nonivamide acetate (SNA) was examined in this present study. Iontophoresis increased the transdermal flux of SNA in vitro as compared to the passive diffusion without any enhancement. Furthermore, iontophoresis was always the most potent enhancement method for SNA permeation among the physical enhancement methods tested. Pulsing of high voltages (electroporation) followed by iontophoresis did not result in increased transport over iontophoresis alone. However, electroporation shortened the onset of transdermal iontophoretic delivery of SNA. Pretreatment of low frequency ultrasound (sonophoresis) alone on skin did not increase the skin permeation of SNA. The combination of iontophoresis and sonophoresis increased transdermal SNA transport more than each method by itself. The enhancement of drug transport across shunt routes and reduction of the threshold voltage in the presence of an electric field may contribute to this synergistic effect. Use of an erbium:YAG laser was a good method for enhancing transdermal absorption of SNA because it allows precise control of stratum corneum (SC) removal, and this ablation of SC could be reversible to the original normal status. The combination of laser treatment and iontophoresis also synergized the skin permeation of SNA, possibly due to a gradual drop in the electric resistance of the skin. The results in this present study point out that the choice of certain conditions with suitable physical enhancement methods can induce a synergistic effect on transdermal delivery of SNA during iontophoresis. PMID- 11879745 TI - Dynamic mechanical thermal analysis studies of polymer films prepared from aqueous dispersion. AB - Dynamic Mechanical Thermal Analysis of cast and sprayed films of an aqueous dispersion of polymethyl methacrylate (Eudragit NE30D) and mixtures with an aqueous dispersion of ethylcellulose (Aquacoat ECD-30) has been undertaken. Such analysis allows the identification of glass transition temperatures and the degree of miscibility of the polymers. It was found that the two polymers formed as cast or sprayed films were not miscible but had an optimal composition of 30% of the ethylcellulose dispersion in the polymethyl methacrylate dispersion. PMID- 11879746 TI - Novel reagents for the sensitive spectrophotometric determination of flutamide, an anticancer drug in pharmaceutical preparations. AB - Simple and sensitive spectrophotometric methods for the determination of flutamide (FLA) in either pure form or in its pharmaceutical preparations are described. The first method is based on the diazotisation of reduced FLA, followed by coupling with alcoholic iminodibenzyl (IDB) in acid medium to give a purple coloured product having a lambda(max) of 570 nm. In the second method, the diazotisation of reduced FLA followed by coupling with 4-amino-5-hydroxy-2,7 naphthalenedisulphonic acid monosodium salt (AHND) in a buffer medium of pH 12, gives a red coloured product having a lambda(max) of 520 nm. Common excipients used as additives in pharmaceutical preparations do not interfere in the proposed methods. Both the methods are highly reproducible and have been applied to a wide variety of pharmaceutical preparations and the results compare favourably with the reported method. PMID- 11879747 TI - Effect of chitosan on a periodontal pathogen Porphyromonas gingivalis. AB - Local delivery systems of antimicrobial agents for treatment of the periodontal diseases received considerable attention during the past decade due to the disadvantages of the systemic administration. An ideal formulation should exhibit ease of delivery, a good retention at the application site, and a controlled release of the drug. The application of bioadhesive gels provides a long stay in the oral cavity, adequate drug penetration, high efficacy and acceptability. In dentistry and oral medicine, various applications of chitosan, which is a bioadhesive polymer have been proposed due to its favorable properties such as biocompatibility and biodegradability. The aim of this study was to determine the antimicrobial activity of chitosan formulations either in gel or film form against a periodontal pathogen, Porphyromonas gingivalis. The viscosity, bioadhesive properties and antimicrobial activity of chitosans at different molecular weight and deacetylation degree were evaluated in the absence or presence of chlorhexidine gluconate (Chx), incorporated into the formulations at 0.1 and 0.2% concentrations. The flow property of the gels were found to be suitable for topical application on the oral mucosa and to syringe into the periodontal pocket. Bioadhesion of the gels and films examined ex-vivo using fresh porcine buccal mucosa showed that both the film and gel formulations exert bioadhesive properties and was not affected by incorporation of Chx. Chitosan is shown to have an antimicrobial activity against P. gingivalis and this was higher with high molecular weight chitosan. The combination of chitosan with Chx showed a higher activity when compared to that of Chx alone, which would provide Chx application at lower concentrations thus avoiding its unwanted side effects. Chitosan films and gels seem to be promising delivery systems for local therapy of periodontal diseases with its bioadhesive property and antimicrobial activity. PMID- 11879748 TI - Preparation, characterization, and tabletting properties of a new cellulose-based pharmaceutical aid. AB - A new cellulose-based tabletting excipient, hereinafter referred to as UICEL, has been developed by treating cellulose powder with an aqueous solution of sodium hydroxide (conc. > or = 5N) and subsequently precipitating it with ethyl alcohol. UICEL is similar in structure to Avicel PH-102, a commercial direct compression excipient commonly referred to as microcrystalline cellulose (MCC). It, however, shows the cellulose II lattice, while Avicel PH-102 belongs to the cellulose I polymorphic form. As produced, UICEL consisted of a mixture of aggregated and non aggregated fibers. The degrees of polymerization (DP) and crystallinity (DC) of UICEL, determined by the viscosity and powder X-ray methods, were 189-207 and 47 58%, respectively. Avicel PH-102, by comparison, showed an aggregated structure with DP and DC values corresponding to 248 and 76.9%, respectively. Compared to Avicel PH-102, UICEL shows higher true density, bulk density, tap density, Carr's index and Hausner ratio values. The mean deformation pressure (P(y)) values calculated from the linear portion of the Heckel plots for UICEL and Avicel PH 102 were about 104 and 87 MPa, respectively, suggesting that UICEL is less ductile than Avicel PH-102. The hardness values of UICEL tablets increased nearly linearly with increasing compression pressures. Comparatively, Avicel PH-102 formed stronger tablets. Irrespective of the compression pressure used, all UICEL tablets disintegrated within 15 s, whereas Avicel PH-102 tablets of comparable strengths remained intact for over 12 h. In conclusion, the results show that UICEL can be used as a direct compression excipient, especially in the design and development of fast-disintegrating tablets. PMID- 11879749 TI - Transdermal delivery of triprolidine using TPX polymer membrane. AB - Triprolidine-containing matrix was fabricated with poly(4-methyl-1-pentene) (TPX) polymer to control the release of the drug. Effect of penetration enhancer and stripping of skin on the permeation of triprolidine through the excised mouse skin was studied. Penetrating enhancers showed the increased flux probably due to the enhancing effect on the skin barrier, the stratum corneum. Among enhancers used such as glycols, fatty acids and non-ionic surfactants, polyoxyethylene-2 oleyl ether showed the best enhancement. The permeability of triprolidine was markedly increased with stripping the mouse skin to remove the stratum corneum, which acts as a barrier of skin permeation. For the controlling delivery of triprolidine, the TPX matrix containing permeation enhancer could be developed. PMID- 11879750 TI - Direct compression properties of melt-extruded isomalt. AB - Isomalt, a sugar alcohol, was melt-extruded prior to compression in order to improve its tabletting properties. After fusion, crystalline isomalt was transformed into an amorphous form as shown by X-ray diffraction and differential scanning calorimetry (DSC). The tabletting properties of amorphous isomalt were dramatically improved. Mixtures formulated with paracetamol (50%) and extruded isomalt yielded hard tablets. However, extruded isomalt powder showed agglomeration problems due to recrystallization of the amorphous phase into a stable crystalline form in the presence of atmospheric moisture. The evolution of the moisture content correlated well with the compressibility data. The tablets made of extruded isomalt powder had a lower friability in comparison to the tablets formulated with non-extruded isomalt powder. Their disintegration was fast and a rapid dissolution rate was recorded. Extruded isomalt displayed excellent tabletting properties; however, further experiments should be conducted to delay or even prevent recrystallization of amorphous isomalt. PMID- 11879751 TI - Direct compression and moulding properties of co-extruded isomalt/drug mixtures. AB - Isomalt, a disaccharide alcohol was co-extruded with paracetamol or hydrochlorothiazide (HCT) in order to improve its tabletting properties. After extrusion, isomalt was transformed into an amorphous form, while paracetamol remained crystalline. Hot stage microscopy showed that HCT was amorphous in the isomalt carrier up to a concentration of 1% (w/w). Direct compression of mixtures formulated with co-extruded isomalt/paracetamol powders yielded harder tablets compared with physical mixtures and no powder agglomeration was observed. Direct moulding of isomalt co-extruded with either paracetamol or HCT was feasible, yielding hard tablets. A fast dissolution rate was seen for both the compressed and the moulded tablets (>80% paracetamol and 60% HCT released within 20 min). The compressed tablets showed a dramatic decrease in tensile strength during storage at 85% RH, while the tensile strength of the moulded tablets remained above 0.80 MPa after 6 months storage at the same conditions. Co-extrusion of isomalt with paracetamol and HCT dramatically improved the tabletting properties of the mixtures (compared with physical mixtures of drug and isomalt). Direct moulding proved to be a suitable technique to produce isomalt based tablets. PMID- 11879752 TI - Dry adsorbed emulsion: 2. Dissolution behaviour of an intricate formulation. AB - The behaviour of a pharmaceutical form, called dry adsorbed emulsion (DAE), containing a sparingly soluble drug (i.e. theophylline) was studied for dissolution drug release kinetic, in relation with DAE structure characterisation. In vitro dissolution testings were performed under different experimental conditions (medium at pH 1.2 and 7.4, medium with or without surfactant addition, different particle sizes, discrete or densified particles). Discrete DAE particles showed an extended release, in comparison with the native drug powder, depending on both drug solubility in the medium and particle size. The relevance of dissolution data was not improved by surfactant addition (0.1% sodium lauryl sulfate: SLS). After an initial release due to theophylline of the DAE superficial layer, the dissolution followed the Higuchi model. This suggested that DAE behaved as an inert matrix, which controlled drug release by diffusion through the hydrophobic part of the DAE. Densified DAE particles showed a slower dissolution rate than discrete DAE particles, because of their weak wettability and their poor disintegrant properties due to the particulate rearrangement under pressure. Lastly in a technological point of view, DAE could be considered as a potential drug delivery system in capsules or tablets to better control bioavailability of drugs. PMID- 11879753 TI - Paclitaxel and its formulations. AB - Paclitaxel (Taxol) is a promising anti-tumor agent with poor water solubility. It is effective for various cancers especially ovarian and breast cancer. Intravenous administration of a current formulation in a non-aqueous vehicle containing Cremophor EL may cause allergic reactions and precipitation on aqueous dilution. Moreover, the extensive clinical use of this drug is somewhat delayed due to the lack of appropriate delivery vehicles. Due to this there is a need for the development of alternate formulation of paclitaxel having good aqueous solubility and at the same time free of any side effects. Various approaches employed so far include cosolvents, emulsions, micelles, liposomes, microspheres nanoparticles, cyclodextrins, pastes, and implants etc. which are discussed in this paper. PMID- 11879754 TI - The effect of spray drying solutions of polyethylene glycol (PEG) and lactose/PEG on their physicochemical properties. AB - The effect of spray drying polyethylene glycol (PEG) 4000 and lactose/PEG solutions was investigated. Micro-spherical PEG particles were successfully prepared from ethanol, which allowed lower outlet temperatures than water. The product was crystalline and consisted of rough spheres or rod like particles. In the case of lactose/PEG composites, spray dried from water, the crystallinity of both components was reduced on spray drying, the extent being dependent on the starting composition. Spray dried lactose/PEG with PEG present as 10% by weight was found to be the most amorphous of the systems prepared. Conversion to more crystalline products occurred over time, the rates of conversion being dependent on temperature and humidity. On storage at low humidity (31-34%) amorphous lactose in lactose/PEG spray dried systems converts to anhydrous crystalline lactose while at high humidity (75% RH) the monohydrate is formed. The rate of transformation of amorphous lactose to the crystalline monohydrate form, at high relative humidity, was quantified using the Avrami equation applied both to X-ray diffraction (XRD) peak intensity and heat of fusion data. Crystallisation of lactose appeared to be retarded at low PEG concentrations, where PEG was present predominantly in a non-crystalline state, but was accelerated at higher PEG contents. PMID- 11879755 TI - Investigation of protein/carbohydrate interactions in the dried state. 2. Diffuse reflectance FTIR studies. AB - Upon freeze-drying in the absence of lyoprotectants, Fourier transform infrared (FTIR) spectroscopy has detected changes in the secondary structures of proteins. Such FTIR studies have been typically conducted using protein/KBr pellets, where additional protein degradation could potentially occur due to pressure effects and partial dissolution of the chaotropic KBr. Diffuse reflectance FTIR spectroscopy, in which no sample preparation is necessary, was evaluated as an alternative spectroscopic method to examine protein structure upon freeze-drying. The therapeutic proteins recombinant human deoxyribonuclease I (rh-DNase) and recombinant human insulin like growth factor I (rh-IGF-I) were freeze-dried with mannitol, sucrose, trehalose, and two molecular weight dextrans (69 and 503 kDa) separately, at concentrations ranging from 0 to 100% (w/w). Upon freeze-drying, rh-DNase and rh-IGF-I underwent significant changes in their secondary structure. For both proteins, the presence of intermolecular beta-sheets due to aggregation was detected and the alpha-helix content decreased significantly. The addition of carbohydrates to the formulations inhibited the protein secondary structure rearrangement in a concentration-dependent manner. Sucrose and trehalose appeared to be the most efficient excipients in preventing secondary structure changes. The conformational changes observed for both proteins appeared to be reversible upon rehydration. PMID- 11879756 TI - Binding of doxycycline to keratin, melanin and human epidermal tissue. AB - Doxycycline is licensed for the prophylaxis of malaria and recent research has indicated the feasibility of delivering this drug across the skin. The binding of doxycycline to keratin could influence skin permeation rates and it has been suggested that the interaction of anti-malarials with melanin may contribute to side effects, such as retinal damage. Doxycycline HCl was incubated with keratin (bovine horn), melanin (Sepia officinalis) and human epidermal samples (native and delipidised). Dose dependent binding of doxycycline to keratin and melanin was observed, and was of similar magnitude for each protein. However, the binding of doxycycline to melanin was lower by an order of magnitude relative to data previously reported for some other anti-malarials, and may indicate reduced side effects. Doxycycline also demonstrated significantly greater affinity for native epidermal skin than for delipidised skin showing that doxycycline, a charged polar molecule, has affinity for the intercellular lipid matrix in addition to the proteinaceous domain. For native skin it was estimated that saturation would be reached at approximately 140 microg x cm(-2); for delipidised skin it was estimated to be 60 microg x cm(-2). Overall, the data suggested that the partition-diffusion steps that are involved in transcellular permeation are possible. PMID- 11879757 TI - Triclosan: release from transdermal adhesive formulations and in vitro permeation across human epidermal membranes. AB - Malarial resistance is an escalating global problem and consequently new and more efficacious treatments to combat malaria are urgently needed. The transdermal delivery of anti-malarials may provide an effective alternative or adjunct to conventional regimens. Triclosan is widely used as an anti-bacterial agent and it has recently been demonstrated that this compound has anti-malarial properties. Its high lipophilicity makes it a potential candidate for delivery across the skin and this paper examines in vitro the potential for the transdermal delivery of triclosan from 'drug-in-glue' formulations. Model patches were prepared using DuroTak 2287, 2516 and 2051 acrylic polymer adhesives loaded with 0, 30 and 50 mg per 0.785 cm(-2) triclosan and dissolution was measured over a 12-h period. There was no apparent difference between the adhesives at the 30 mg patch loading, but at 50 mg, the trend for increased release was 2051>2516>2287. No significant burst effect was apparent. Patches of 50 mg per 0.785 cm2 were then used to determine the permeation of triclosan across heat-separated human epidermal membranes in Franz diffusion cells, over a period of 48 h. The above general trend was reflected in the steady state flux values obtained: 2051:16.91 microg x cm(-2) x h(-1) (S.E.M. 1.29), 2516:15.05 microg x cm(-2) x h(-1) (S.E.M. 1.00), 2287 12.83 microg x cm(-2) x h(-1) (S.E.M. 2.81). Although pharmacokinetic data are not currently available to permit calculation of an efficacious patch size, the transdermal delivery of triclosan is feasible. PMID- 11879758 TI - Effects of formulation variables on characteristics of poly (ethylcyanoacrylate) nanocapsules prepared from w/o microemulsions. AB - The effect of several formulation variables on some of the physico-chemical characteristics of poly (ethyl cyanoacrylate) (PECA) nanocapsules prepared by the interfacial polymerisation of biocompatible water-in-oil microemulsions was investigated. In all cases, yields were high (>90%) and the polydispersity in size of nanocapsules was narrow. The molecular weight of the nanocapsules formed was influenced by the pH of the aqueous component of the microemulsion, increasing with increasing pH. The size of the nanocapsules formed (ranging from around 130 to 180 nm) was a function of the ratio of the mass of monomer used to the water weight fraction of the microemulsion, increasing as this ratio was increased. This is due to the formation of a thicker polymer wall resulting from the increased mass of monomer available per unit interfacial area as this ratio is increased. The rate of release of insulin from nanocapsules was also influenced by this ratio, in agreement with its effect on wall thickness. This study demonstrates that many pharmaceutically relevant physico-chemical properties of poly (alkyl cyanoacrylate) (PACA) nanocapsules prepared by interfacial polymerisation of microemulsions can readily be manipulated by changing either the pH of the aqueous component, the water weight fraction of the microemulsion or the mass of monomer used for polymerisation. PMID- 11879759 TI - Preparation and in vitro characterization of a eutectic based semisolid self nanoemulsified drug delivery system (SNEDDS) of ubiquinone: mechanism and progress of emulsion formation. AB - The objectives of the present work were, first, to develop a self-nanoemulsified drug delivery system (SNEDDS) based on the eutectic properties of ubiquinone (CoQ10); and second, to study the progress of emulsion formation and drug release mechanisms by turbidimetry and droplet size analysis. Binary phase diagrams of CoQ10 with menthol and essential oils were constructed and used to develop the self-nanoemulsified formulation. Pseudo ternary phase diagram was constructed to identify the efficient self-emulsification region. Release mechanisms of the resultant formulas were quantified using turbidimetry in combination with dissolution studies. Turbidity time profiles revealed three distinctive regions: lag phase, plateau, and the pseudolinear phase. Lag phase was attributed to the liquid crystalline properties of the formula. Plateau turbidity was correlated with droplet size. Laser diffraction analysis revealed an average droplet diameter of 100 nm. Emulsification rate was obtained from the corrected slope of the pseudolinear phase of the profile. Stability of the formula was further evaluated using Fourier transform-infrared (FT-IR) attached to an attenuated total reflectance (ATR) accessory. The present study revealed a eutectic based semisolid self-emulsified delivery system that can overcome the drawbacks of the traditional emulsified systems such as low solubility and irreversible precipitation of the active drug in the vehicle with time. PMID- 11879760 TI - Outreach: targeting high-risk women through community partnerships. AB - The need for community-based preventive intervention is driven by the ineffectiveness and the costliness of traditional approaches. We argue that community outreach efforts should be characterized by three components: 1) careful mapping of high-risk areas, 2) developing partnerships with trusted community institutions within areas of high risk, and 3) developing a portfolio of institutional partners that maximize the penetration of high-risk populations. The analysis of these high-risk contexts redirects the focus from individuals to institutional structures. Gaining a greater understanding of how impoverished women relate to formal institutions is critical to the primary goal of reducing unnecessary deaths. PMID- 11879761 TI - Contraceptive counseling in managed care: preventing unintended pregnancy in adults. AB - This study examines contraceptive counseling received by adult women in their managed care plans and the relationship between counseling and women's contraceptive attitudes and practices. Telephone interviews were conducted with a random sample of 898 women ages 18 to 44 enrolled in a commercial health maintenance organization (HMO) or point-of-service (POS) health plan. Counseling received in the past 2 years was measured on three dimensions: exposure through any communication channel; content of information; and personalization of discussion. Multiple logistic regression analysis was used to examine the determinants of counseling and the relationship between counseling and four outcomes: satisfaction with counseling received, self-efficacy for preventing unintended pregnancy, current use of contraception (if at risk of unintended pregnancy), and intent to contracept in the next year (if at risk). Overall, 60.5% of women were at risk of unintended pregnancy; among those at risk, 69% received any counseling in the past 2 years, compared with 38% among those not at risk. Receiving personalized counseling (as opposed to no counseling or only informational counseling) significantly increases the odds of satisfaction with counseling, current contraceptive use, and intent to contracept. Informational counseling alone (without personalization) significantly increases the odds of contraceptive use. Women ages 40-44 were less likely than younger women to receive counseling and to use contraception if at risk of unintended pregnancy. We conclude that receiving contraceptive counseling in managed care is associated with contraceptive attitudes and practices among adults and that there is substantial room for quality improvement in the provision of contraceptive counseling. PMID- 11879762 TI - Preventive health services use, lifestyle health behavior risks, and self reported health status of women in Ohio by ethnicity and completed education status. AB - This study assessed the health status and behavior of college-educated and non college-educated African American women and European American women in Ohio. Analyses focused on health services utilization, health status, and life style/health behaviors from the 1998 Ohio Family Health Survey. College-educated African American women used more preventive health services and had better health status than non-college-educated African American women. Even so, college educated African American women still had higher body mass index values, lower health status, and higher percent currently smoking than college-educated European American women. We conclude that college-educated African American women may face unique barriers to implementing all types of health-promoting behaviors available consequent to their higher education. Partnerships with respected community institutions, such as churches, may help these women develop good health practices in their entire community as well as in themselves. PMID- 11879763 TI - The delivery of medical abortion services: the views of experienced providers. AB - This study examined beliefs about the delivery of medical abortion services and current roles of 76 providers of methotrexate-induced abortion. The sample included physicians, midlevel professionals, administrators, and counselors/other medical staff. Over 75% of participants believed that, given proper training and emergency backup, physicians and midlevel clinicians alike could provide medical abortions. Over 85% agreed that methotrexate-induced abortions could be provided in any physician's office or medical facility. There were no differences in perceptions by participant group. Involvement of midlevel providers in provision of medical abortion could potentially increase access and options for women seeking to terminate an early pregnancy. PMID- 11879764 TI - Fly-ing from genes to drugs. PMID- 11879765 TI - Functions of the fragile X protein. PMID- 11879766 TI - From genomes to cures: the start of a long journey. AB - The EMBO/EMBL Science and Society Conference 'From Genomes to Cures', was held in Heidelberg, Germany, 16-18 November 2001. PMID- 11879770 TI - Microchimerism: incidental byproduct of pregnancy or active participant in human health? AB - It is now well recognized that cells traffic in both directions between fetus and mother during pregnancy. Moreover, fetal cells have been found to persist for years, probably for a lifetime, in the circulation of healthy women. Harboring of cells from another individual at low levels is called microchimerism. Women have a predilection to autoimmune disease, and chronic graft-versus-host disease, a condition of human chimerism, shares similarities with some autoimmune diseases. The specific HLA genes of donor and host are known to be of central importance in graft-versus-host disease, and HLA class II genes are important in autoimmune disease. Considered together, these observations led to the hypothesis that microchimerism and HLA genes of host and non-host cells are involved in autoimmune diseases. Alternative sources of microchimerism include transfer from a twin or the mother during pregnancy, or from blood transfusion. Studies of systemic sclerosis, primary biliary cirrhosis, Sjogrens syndrome, pruritic eruption of pregnancy, myositis, and thyroid disease have both lent support and raised doubts about a potential role of microchimerism in autoimmune disease. PMID- 11879771 TI - Medicinal chemistry and therapeutic potential of CpG DNA. AB - The observation that oligodeoxynucleotides containing CpG dinucleotides (CpG DNA) exhibit several immunological effects has led to their use as therapeutic agents and adjuvants for various diseases. Several CpG DNA drug candidates are currently being evaluated, either as monotherapies or as adjuvants (with vaccines, antibodies, antigens and allergens), in preclinical and clinical trials against cancers, viral and bacterial infections, allergies and asthma. Knowledge gained from studies of the medicinal chemistry of CpG DNA has provided a basis for designing a second generation of CpG DNA agents with desirable cytokine-inducing and potent immunomodulatory activity. This article reviews recent progress in understanding the effects of CpG DNA, the medicinal chemistry of CpG DNA, and its possible therapeutic applications. PMID- 11879772 TI - New immunotherapeutic strategies to control vaginal candidiasis. AB - The widespread occurrence of mucosal infections caused by Candida, in particular recurrent vulvovaginal candidiasis among fertile-age women, together with the paucity of safe candidacidal antimycotics, have prompted a great number of investigations into the immunotherapy of candidal vaginitis. This article will discuss three different experimental approaches demonstrated to be potentially transferable to human disease: (1) the use of antibodies against well-defined cell-surface adhesins or enzymes; (2) the generation of yeast killer-toxin-like candidacidal anti-idiotypic antibodies and their engineered molecular derivatives (e.g. single chains, peptides); and (3) the generation of therapeutic vaccines and immunomodulators. PMID- 11879773 TI - Glycogen synthase kinase 3: an emerging therapeutic target. AB - Glycogen synthase kinase 3 (GSK-3) is a serine/threonine protein kinase that has recently emerged as a key target in drug discovery. It has been implicated in multiple cellular processes and linked with the pathogenesis of several diseases. GSK-3 inhibitors might prove useful as therapeutic compounds in the treatment of conditions associated with elevated levels of enzyme activity, such as type 2 diabetes and Alzheimer's disease. The pro-apoptotic feature of GSK-3 activity suggests a potential role for its inhibitors in protection against neuronal cell death, and in the treatment of traumatic head injury and stroke. Finally, selective inhibitors of GSK-3 could mimic the action of mood stabilizers such as lithium and valproic acid and be used in the treatment of bipolar mood disorders. PMID- 11879774 TI - Mutations in the p53 homolog p63: allele-specific developmental syndromes in humans. AB - p63 is the most recently discovered but most ancient member of the p53 family. In marked contrast to p53, p63 is highly expressed in embryonic ectoderm and in the basal, regenerative layers of many epithelial tissues in the adult. The p63 knockout mouse dies at birth and lacks limbs, epidermis, prostate, breast and urothelial tissues, apparently owing to the loss of stem cells required for these tissues. Significantly, several dominant human syndromes involving limb development and/or ectodermal dysplasia have been mapped to chromosome 3q27 and ultimately the gene encoding p63. The heterozygous p63mutations are distinct for each of the syndromes and are thought to act through both dominant-negative and gain-of-function mechanisms rather than a loss-of-function haploinsufficiency. The allele specificity of these syndromes offers unique molecular insights into the poorly understood actions of p63 in limb development, ectodermal-mesodermal interactions and stem cell maintenance. PMID- 11879776 TI - Embryo and stem cell research in the USA: a political history. AB - Embryo and stem cell research in the USA is inextricably linked with the abortion debate. Since 1973, when elective abortions became legal, the government has refused to fund therapeutic research for fear it would encourage abortions. Research has continued in an unregulated private sector. Here, the unique US cultural, political and religious factors that have resulted in this situation are discussed. PMID- 11879775 TI - Disease model: Fanconi anemia. AB - Fanconi anemia (FA) is a chromosomal instability syndrome characterized by the presence of pancytopenia, congenital malformations and cancer predisposition. Six genes associated with this disorder have been cloned, and mice with targeted disruptions of several of the FA genes have been generated. These mouse models display the characteristic FA feature of cellular hypersensitivity to DNA cross linking agents. Although they do not develop hematological or developmental abnormalities spontaneously, they mimic FA patients in their reduced fertility. Studies using these animal models provide valuable insights into the involvement of apoptotic pathways in FA, and help characterize the defects in FA hematopoietic cells. In addition, mouse models are also useful for testing treatments for FA. PMID- 11879779 TI - Temporal expression of fumonisin B(1)-induced tumor necrosis factor-alpha and interferon gamma in mice. AB - Fumonisin B(1) (FB(1)), a toxic metabolite of Fusarium verticillioides, is a carcinogen and causative agent of various animal diseases. Our previous studies indicated the involvement of tumor necrosis factor-alpha (TNF alpha) in FB(1) induced toxic responses. To further investigate the time-course of TNF alpha production and signaling, mice (four/group) were treated subcutaneously (s.c.) or per os (p.o.) with either vehicle or 25 mg/kg of FB(1) as a single dose and sacrificed at 0, 2, 4, 8, 12 and 24 h after treatment. The TNF alpha expression was increased in liver and kidney after both routes of FB(1) exposure without any alterations in spleen. The p.o.-route FB(1) treatment caused greater hepatotoxicity compared to the s.c. route, as depicted by increased alanine aminotransferase and aspartate aminotransferase level in plasma, observed only after p.o. FB(1) treatment. The increase in enzymes at 8 h after p.o. treatment correlated with the highest TNF alpha expression, also noted at 8 h after p.o. treatment, thus further confirming the involvement of TNF alpha in FB(1) toxicity. The interferon (IFN)-gamma expression was increased in liver at 4 h after p.o. FB(1) treatment, suggesting a possible combined role of TNF alpha and IFN gamma in their induction and hepatotoxicity. PMID- 11879781 TI - Seasonal variations of pollution biomarkers in two populations of Corbicula fluminea (Muller). AB - Seasonal variations of pollution biomarkers have been studied in two populations of Corbicula fluminea (Muller) for 2 years. Clams were collected from the Sanguinet lake and the Dronne river (Southwest France), sites characterised by water of excellent and passable quality, respectively. Biological parameters studied were enzymatic activity (catalase, propionylcholinesterase, glutathione S transferase, NADPH-cytochrome c reductase and NADH-cytochrome c reductase activities) and indicators of physiological (protein, carbohydrate, lipid and ash contents, fresh and dry tissue weights, condition factor) and reproductive (amount of oocytes present in gonads and clams incubating larvae in gills) status. Environmental parameters (e.g. water temperature, chlorophyll a concentration) were also monitored. Effects of site, sampling date and season (from spring to winter) were studied, as well as relationships between abiotic and biotic factors. Finally, an index of variability was calculated for each biomarker. Catalase, propionylcholinesterase, NADH-cytochrome c reductase and lipids experienced the most pronounced seasonal fluctuations, whereas glutathione S-transferase, NADPH-cytochrome c reductase and proteins suffered less pronounced ones. For some parameters, results differed depending on the site. Most seasonal changes were related to the reproductive cycle and/or water temperature. Implications of such variations in the use of pollution biomarkers in environmental surveys are discussed. PMID- 11879780 TI - Induction of a putative metallothionein gene in the blood cockle, Anadara granosa, exposed to cadmium. AB - The relationship between a putative metallothionein gene (MT) and exposure to cadmium (Cd) in blood cockles (Anadara granosa) is reported. In a 96-h dose response experiment, mortality of cockles was found to proportionately increase in the range of 0.2-5.0 mg/l Cd with a calculated LC(50) of 2.94 mg/l. Exposure to 0.25 mg/l Cd for 16 days caused significant increases (P<0.05) in Cd concentrations in whole tissues, gills and hepatopancreas, and the accumulation of Cd in these tissues increased with the duration of exposure. Two cDNA libraries constructed using the hepatopancreas from control and Cd-treated cockles gave titres of 5.62 x 10(5) and 1.94 x 10(5) pfu/microg vector, respectively. A putative MT gene, AnaMT, of 510 nucleotides in length, was isolated from the treated cDNA library using a heterologous probe MT20 from the blue mussel, Mytilus edulis. Northern analyses using AnaMT as a probe indicated low expression of the MT mRNA in control animals. In cockles treated with 0.25 mg/l Cd for 4 days, MT mRNA level increased to approximately 168%, but declined to 108% at day 8. After 12 and 16 days of Cd treatment, expression of the MT gene was 138% and 187%, respectively, compared to the controls. These observations suggest that induction of the MT gene by a sublethal dose of Cd is rapid, occurring within 4 days of treatment. PMID- 11879782 TI - Calcium involvement in the luminescence control of three ophiuroid species (Echinodermata). AB - Although it has been shown that calcium is involved in the control of the luminous reaction of many invertebrate phyla, its role in Echinoderms is poorly documented. The aim of this work was to carry out a comparative study of calcium requirement of KCl-induced light emission by arm segments and dissociated luminous cells from three ophiuroid species, Ophiopsila californica, O. aranea and Amphiura filiformis. Results show a gradual inhibition of the luminescence when preparations are incubated in artificial sea water with lowered calcium concentration. The calcium substitutes Ba(2+) and Sr(2+) could act either as blockers or as substitutes, depending on the ophiuroid species; while calcium blockers Co(2+), Ni(2+) and Cd(2+) inhibit light emission in A. filiformis and in O. californica, but not in O. aranea. The nature of putative calcium voltage gated channel has been studied pharmacologically using 1,4-dihydropyridine, benzodiazepine, phenylalkylamine and trifluoroperazine. From our results, it is proposed that calcium could act via an L-type voltage-gated calcium channel in O. californica and A. filiformis but not in O. aranea. The precise role of calcium in luminescence control still remains unknown; it could act as a second messenger or as a co-factor of the luminous reaction. PMID- 11879783 TI - Modulation of rat liver cytochrome P450 activity by prolonged red wine consumption. AB - Cytochrome P450-dependent oxidation of lauric acid, p-nitrophenol and ethanol by liver microsomal fractions were studied in control rats and in animals given either ethanol, red wine, or alcohol-free red wine for 10 weeks. Ethanol increased the total cytochrome P450 and the isoenzyme 2E1 content, as well as the p-nitrophenol hydroxylation and ethanol oxidation. These effects of ethanol treatment were attenuated by red wine administration. Red wine increased the total antioxidant capacity of plasma, whereas the alcohol-free red wine decreased the cytochrome P450 content and decreased the oxidation of lauric acid, p nitrophenol and ethanol to values lower than control. It is concluded that red wine administration attenuates the ethanol-induced enhancement in liver microsomal parameters dependent on cytochrome P450 2E1 activity, an affect that seems to be accomplished by the non-alcoholic constituents of red wine known to have antioxidant properties. PMID- 11879784 TI - Membrane effects of toxins isolated from a cyanobacterium, Cylindrospermopsis raciborskii, on identified molluscan neurones. AB - The effect of anatoxin (ANTX), the crude extract (AlgTX) and purified fraction (F1) isolated from cyanobacterium C. raciborskii was studied on the neurones of two snail species. ANTX and AlgTX exerted excitatory, inhibitory and biphasic effects on the spontaneous activity of identified neurones. Both ANTX and AlgTX elicited an inward current, which could be decreased by curare or amiloride. On the contrary, F1 had no direct effect on the spontaneous activity; it was not able to induce conductance changes of the neuronal membrane, but it did antagonise the acetylcholine (ACh)-induced inward current. We concluded that ANTX affects the neuronal membrane of neurones acting on ACh receptors. The AlgTX had similar effects, and therefore the extract of C. raciborskii may contain an ANTX like component. The purified fraction prolonged and decreased the ACh-elicited response, but had no direct membrane effect. We suggest, therefore, that both AlgTX and the purified fraction F1 interact with the ACh receptor, but they have different binding sites on the neuronal ACh receptor-ion channel complex. The possible neurotoxic effects of the C. raciborskii extract and F1 are demonstrated for the first time; the molecular mechanism of their action, however, remains to be elucidated. PMID- 11879785 TI - Enhanced heme oxygenase activity increases the antioxidant defense capacity of guinea pig liver upon acute cobalt chloride loading: comparison with rat liver. AB - Changes in the activity of so-called oxidative stress defensive enzymes, superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase and heme oxygenase, as well as changes in lipid peroxidation and reduced glutathione levels, were measured in guinea pig and rat liver after acute cobalt loading. Cobalt chloride administration produced a much higher degree of lipid peroxidation in guinea pig than in rat liver compared with the control animals. The intrahepatic reduced glutathione content in control guinea pig was higher than that in rat, but was equally decreased in both species after cobalt administration. The enzymatic scavengers of free radicals, superoxide dismutase, catalase and glutathione peroxidase, were significantly decreased in rat liver after acute cobalt loading, and as a compensatory reaction, the heme oxygenase activity was increased (seven-fold). In guinea pig liver, only superoxide dismutase activity was depleted in response to cobalt-induced oxidative stress, while catalase and glutathione peroxidase were highly activated and the heme oxygenase activity was dramatically increased (13-fold). It is assumed that enhanced heme oxygenase activity may have important antioxidant significance by increasing the liver oxidative-stress defense capacity. PMID- 11879787 TI - Antioxidant systems of the developing quail embryo are compromised by mycotoxin aurofusarin. AB - The effects of aurofusarin in the quail diet on the antioxidant systems of the developing embryo are investigated. Thirty eight 45-day-old Japanese quails (Coturnix japonica) were divided into two groups and were fed on a corn-soya diet or the same diet supplemented with aurofusarin at the level of 26.4 mg/kg feed in the form of Fusarium graminearum culture enriched with aurofusarin. Eggs obtained after 7 weeks of feeding were incubated. Samples of quail tissues were collected at day 17 of embryonic development and from day old hatchlings. Antioxidants and malondialdehyde were analysed by HPLC-based methods. Inclusion of aurofusarin in the maternal diet was associated with decreased concentrations of alpha- and gamma-tocopherols, alpha- and gamma-tocotrienols, retinol, lutein and zeaxanthin in egg yolk. The vitamin E (tocopherols and tocotrienols) concentration in the liver and yolk sac membrane (YSM) of the day 17 embryos and the hatchlings from aurofusarin-fed group was significantly decreased. Alpha-tocopherol concentration was also reduced in kidney, lung, heart, muscle and brain of day-old quails. In the liver of day-old quails, concentrations of lutein, zeaxanthin, retinol, retinyl linoleate, retinyl oleate, retinyl palmitate and retinyl stearate were also reduced. As a result of these diminished antioxidant concentrations, tissue susceptibility to lipid peroxidation was significantly increased. It is suggested that a compromised antioxidant system of the egg yolk and embryonic tissues could predispose quails to increased mortality at late stages of their embryonic development. PMID- 11879786 TI - Effect of ligands and other metals on the uptake of mercury and methylmercury across the gills and the intestine of the blue crab (Callinectes sapidus). AB - Using the perfusion method, we compared the accumulation and flux of inorganic mercury (Hg) and methylmercury (CH(3)Hg) across the gills and intestine of the blue crab, Callinectes sapidus. The accumulation and transfer processes were studied for each form by exposing the organs in the presence of specific ligands and other metals. While binding of Hg and CH(3)Hg to organic ligands reduced the rate of uptake in most instances, the differences in accumulation could not be explained only in terms of passive diffusive uptake. Thus, it appears that Hg and CH(3)Hg accumulation is dominated by ligand exchange or facilitated transport processes. Exposure of the gills and intestine in the presence of a suite of metals and metalloids showed that inorganic Hg and CH(3)Hg uptake was largely by different mechanisms to that of the other elements, as there was little interaction in terms of uptake rate. Overall, the results of this study suggest that inorganic Hg and CH(3)Hg uptake into the gills and intestine of this invertebrate is by a variety of pathways, both active and passive. PMID- 11879788 TI - Comparative study of the purification and characterization of the cytosolic glutathione S-transferases from two salmonid species: Atlantic salmon (Salmo salar) and brown trout (Salmo trutta). AB - In the present report, an efficient method for isolating multiple cytosolic forms of glutathione S-transferases from liver and kidney cytosolic samples of two salmonid species (brown trout and Atlantic salmon) is described, and some of the multiple properties of these enzymes are presented. Glutathione S-transferases were partially purified by low-pressure affinity chromatography on a column with glutathione coupled to agarose, which retained an average of 89.47% of the total activity. The GST activity was appropriated towards CDNB and ETHA as substrates. The application of an HPLC system associated to elestrospray ionization mass spectrometry allowed the identification of five GST cytosolic isoforms, corresponding to subunits with M(r) between 23,700 and 26,900 Da being the main form, with retention time of 17 min, a pi-class-related GST isoenzyme. PMID- 11879789 TI - Non-granule PSA-NCAM immunoreactive neurons in the rat hippocampus. AB - The polysialylated form of the neural cell adhesion molecule (PSA-NCAM) continues to be expressed in the adult hippocampus, mainly in a subset of neurons located in the innermost portion of the granule cell layer. PSA-NCAM immunoreactive neurons have also been described outside this layer in humans, where they are severely reduced in schizophrenic brains. Given this important clinical implication, we were interested in finding whether similar neurons existed in the adult rat hippocampus and to characterize their distribution, morphology and phenotype. PSA-NCAM immunocytochemistry reveals labeled neurons in the subiculum, fimbria, alveus, hilus, and stratum oriens, lucidum and radiatum of CA3 and CA1. They are mainly distributed in the ventral hippocampus, and have polygonal or fusiform somata with multipolar or bipolar morphology. These neurons show long straight dendrites, which reach several strata and even enter the fimbria and the alveus. These dendrites are often varicose, appear devoid of excrescences and apparently do not show spines. Most of these neurons display GABA immunoreactivity and further analysis has shown that a subpopulation expresses calretinin, but not somatostatin, neuropeptide Y, parvalbumin, calbindin or NADPH diaphorase. Our study demonstrates that there is an important subpopulation of PSA-NCAM immunoreactive neurons, many of which can be considered interneurons, outside the rat granule cell layer, probably homologous to those described in the human hippocampus. The presence of the polysialylated form of NCAM in these neurons could indicate that they are undergoing continuous remodeling during adulthood and may have an important role in hippocampal structural plasticity. PMID- 11879790 TI - Rat strain differences in nicotine self-administration using an unlimited access paradigm. AB - An effective animal model for elucidating the neurobiological basis of human smoking should simulate important aspects of this behavior. Therefore, a 23 h unlimited access nicotine self-administration model was used to compare inbred Lewis rats, which have a propensity to self-administer drugs of abuse, to inbred Fisher 344 rats and to the outbred Holtzman strain. Using this unlimited access model, 88.8% of Lewis vs. 57.1% of Holtzman rats achieved maintenance self administration at a fixed ratio 1 (FR 1) at 0.03 mg/kg IV nicotine (P<0.05). In contrast, Fisher rats did not acquire self-administration under these conditions. Of the Lewis and Holtzman rats that achieved maintenance self-administration on an FR 1 schedule, a greater percentage of Lewis rats acquired nicotine self administration at FR 2 (P<0.05) and progressed to FR 4 (P<0.05). Using naive cohorts in a progressive dose reduction study, 83.3% of Lewis rats achieved maintenance at 0.0075 mg/kg nicotine as compared to 31.8% of Holtzman rats (P<0.05). Furthermore, only Lewis rats showed differences in active vs. inactive bar presses during maintenance at sequential dose reductions (P<0.001). Thus, in this unlimited access model, inbred Lewis rats will more reliably acquire nicotine self-administration than outbred Holtzman rats. Moreover, Lewis rats showed a significantly higher likelihood of continuing to self-administer nicotine in face of both increasing work requirements and decreasing drug reinforcement. Therefore, it is likely that Lewis rats would be genetically susceptible to nicotine addiction. PMID- 11879791 TI - Long-term effects of St. John's wort and hypericin on monoamine levels in rat hypothalamus and hippocampus. AB - Hypericum perforatum L. (St. John's wort) is one of the leading psychotherapeutic phytomedicines and, because of this, great effort has been devoted to clarifying its mechanism of action. Chronic effects of St. John's wort and hypericin, one of its major active compounds, on regional brain amine metabolism have not been reported yet. We used a high-performance liquid chromatography system to examine the effects of short-term (2 weeks) and long-term (8 weeks) administration of imipramine, Hypericum extract or hypericin on regional levels of serotonin (5 HT), norepinephrine, dopamine and their metabolites in the rat brain. We focused our interest on the hypothalamus and hippocampus, as these brain regions are thought to be involved in antidepressant drug action. Imipramine (15 mg/kg, p.o.), Hypericum extract (500 mg/kg, p.o.), and hypericin (0.2 mg/kg, p.o.) given daily for 8 weeks significantly increased 5-HT levels in the hypothalamus (P<0.05). The 5-HT turnover was significantly lowered in both brain regions after 8 weeks of daily treatment with the Hypericum extract (both P<0.05). Consistent changes in catecholamine levels were only detected in hypothalamic tissues after long-term treatment. Comparable to imipramine, Hypericum extract as well as hypericin significantly decreased 3,4-dihydroxyphenylacetic acid and homovanillic acid levels in the hypothalamus (P<0.01). Our data clearly show that long-term, but not short-term administration of St. John's wort and its active constituent hypericin modify levels of neurotransmitters in brain regions involved in the pathophysiology of depression. PMID- 11879792 TI - The ability of amphetamine to evoke arc (Arg 3.1) mRNA expression in the caudate, nucleus accumbens and neocortex is modulated by environmental context. AB - The ability of amphetamine or cocaine to induce the expression of c-fos mRNA in a number of brain regions is greatly enhanced when these drugs are administered in a distinct and relatively novel environment, relative to when they are given in the home cage. The purpose of this study was to determine if environmental context has a similar effect on the ability of amphetamine to induce the expression of arc (also known as Arg 3.1), an "effector" immediate early gene (IEG) thought to play a direct role in cellular plasticity. Rats were administered either saline or amphetamine (0.5 mg/kg, i.v.), in their home cage or in a distinct test environment. Fifty minutes later, they were decapitated and their brains processed for in situ hybridization histochemistry. In the prefrontal cortex, caudate-putamen and core of the nucleus accumbens, amphetamine significantly increased arc mRNA expression under both conditions, but the level of expression was significantly enhanced when amphetamine was given in a distinct environment. In the shell of the nucleus accumbens amphetamine significantly increased the expression of arc mRNA only when it was administered in the distinct environment. Thus, the ability of amphetamine to induce the expression of arc varies as a function of the environmental context in which it is administered. This could contribute to the ability of environmental context to modulate forms of drug experience-dependent neuroplasticity, including behavioral sensitization. PMID- 11879793 TI - Central urocortin activation of sympathetic-regulated energy metabolism in Wistar rats. AB - The corticotropin-releasing hormone (CRH) system, including CRH and urocortin (UCN), is implicated in the central control of appetite and energy metabolism. Urocortin, a recently isolated neuropeptide closely related to CRH is involved in the central signaling cascade that inhibits energy intake. When administered intracerebroventricularly and intra-hypothalamically, UCN potently decreases food intake. Receptors for UCN, while widely distributed, are expressed in hypothalamic nuclei. As the hypothalamus is involved in modulating autonomic outflow, UCN may also act as a catabolic neuropeptide to facilitate energy expenditure through sympathetic-regulated thermogenesis. To test the hypothesis that UCN also enhances regulatory energy expenditure via the activation of the sympathetic nervous system, we examined whole body oxygen consumption (VO(2)) and colonic temperature in male Wistar rats in response to central UCN administration. That is, the intracerebroventricular injection of 1.0 microg of UCN in male Wistar rats (n=10) significantly increased whole body oxygen consumption compared to PBS control. In addition, colonic temperature was significantly increased (Delta0.7 +/- 0.08 degrees C) in UCN- vs. PBS administered rats, which was prevented by pretreatment with the ganglionic blocker chlorisondamine. These studies suggest that UCN acutely increased whole body oxygen consumption and body temperature via central activation of sympathetic outflow. PMID- 11879794 TI - Influence of the hyperpolarization-activated cation current, I(h), on the electrotonic properties of the distal apical dendrites of hippocampal CA1 pyramidal neurones. AB - The electrical field application technique has revealed that the electrotonic length of the distal apical dendrites of hippocampal CA1 pyramidal neurones is long compared to the rest of the cell. This difference may be due to an asymmetrical distribution of channels responsible for the leak conductance in distal and proximal membrane segments. One such conductance, the hyperpolarization-activated cation current, I(h), is reported to display an increasing density with distance from the soma along the apical dendrite. Such asymmetry of I(h) could be a major cause of the increased electrotonic length of the distal apical dendrite. In the present study we found that blockade of I(h), by bath application of Cs(+) (3 mM) or ZD7288 (20 microM), reduced the electrical field-induced transmembrane polarization (TMP) in the distal apical dendrites. In some neurones the polarization reversed polarity, reflecting a movement of the indifference point (site of zero polarization) from the distal dendrites, across the recording site to a more proximal position. These effects were more pronounced when Cs(+) and ZD7288 were applied locally to the distal apical dendrites. Bath application of another antagonist of leak conductance, Ba(2+) (1 mM), also decreased the average field-induced polarization. This latter effect, however, did not reach statistical significance. These data suggest that I(h) is partly responsible for the distal location of the indifference point, and indicate that an elevated activity of I(h) contributes to the relatively increased electrotonic length of the most distal part of the apical dendrites. PMID- 11879795 TI - Motoneuron morphological alterations before and after the onset of the disease in the wobbler mouse. AB - The wobbler mutant mouse displays a recessively inherited neurological disease with degeneration of motoneurons and is considered to be an animal model for human motoneuron diseases. Mutant mice can be clinically recognised at about 3-4 weeks of age but a polymorphic marker close to the wobbler gene offers the opportunity of a preclinical diagnosis. Using this polymorphic marker we performed morphometric (cell size) analysis of spinal cord motoneurons from 10 to 40 days post natal (PN). We observed at day 16 PN a transient appearance of swollen motoneurons, probably those that present vacuolar degeneration a little later and possibly die. One week later, from 21 days onwards, we found that the subpopulation of large motoneurons was depleted in the mutant mice. The absence of large motoneurons may have important physiological consequences and the loss or absence of differentiation of this particular subpopulation of motoneurons may be a key event in the course of the disease. PMID- 11879796 TI - Endogenous parathyroid hormone-related protein functions as a neuroprotective agent. AB - Parathyroid hormone-related protein (PTHrP) was discovered a dozen years ago as a product of malignant tumors. It is now known that PTHrP is a paracrine factor with multiple biological functions. One such function is to relax smooth muscle by inhibiting calcium influx into the cell. In the central nervous system, PTHrP and its receptor are widely expressed in neurons in the cerebral cortex, hippocampus and cerebellum. The function of PTHrP in the CNS is not known. Previous work has shown that expression of the PTHrP gene is depolarization dependent in cultured cerebellar granule cells and depends specifically on L-type voltage sensitive calcium channel (L-VSCC) Ca(2+) influx. PTHrP has also been found to be capable of protecting these cells against kainic acid-induced excitotoxicity. Here, we tested the idea that mice with a PTHrP-null CNS might display hypersensitivity to kainic acid excitotoxicity. We found that these mice were six-fold more sensitive than control littermate mice to kainic-acid-induced seizures as well as hippocampal c-Fos expression. PTHrP-null embryonic mixed cerebral cortical cultures were more sensitive to kainic acid than control cultures, and PTHrP addition was found to be protective against kainate toxicity in both PTHrP-null and control cultures. By whole-cell techniques, PTHrP was found to reduce L-VSCC Ca(2+) influx in cultured mouse neuroblastoma cells. We conclude that PTHrP functions as a component of a neuroprotective feedback loop that is structured around the L-type calcium channel. This loop appears to be operative in vivo as well as in vitro. PMID- 11879797 TI - Antibody directed against GD(2) produces mechanical allodynia, but not thermal hyperalgesia when administered systemically or intrathecally despite its dependence on capsaicin sensitive afferents. AB - Anti-GD(2) antibodies have been shown to be effective for immunotherapy of neuroblastoma and other GD(2) enriched malignancies. Infusion of anti-GD(2) antibodies frequently causes spontaneous pain and allodynia for the duration of the immunotherapy and occasionally longer lasting neuropathic pain. Bolus intravenous injection of anti-GD(2) in rats initiates mechanical allodynia as measured by withdrawal threshold of the hindpaws. In this study, thermal thresholds were measured prior to and for up to 6 h following systemic anti-GD(2) administration in adult rats. In addition, both thermal and mechanical thresholds were tested following intrathecal administration of anti-GD(2) and IgG(2a). Murine anti-GD(2) elicited mechanical allodynia when administered into either the vasculature or the intrathecal space. Effective systemic doses were 1--3 mg/kg as previously shown. Intrathecally, optimal doses ranged from 0.01 to 0.1 ng; a higher dose was ineffective. Thermal hyperalgesia was not observed via either route of administration. Intrathecal pretreatment 48--72 h prior to the experiment with capsaicin at doses sufficient to cause a 50% depletion of dorsal horn CGRP, caused a total blockade of the mechanical allodynia indicating an involvement of peptidergic fine afferent fibers. It is likely that the antibody reacts with an antigen on peripheral nerve and/or myelin to initiate its effect. The lack of observed thermal hyperalgesia is surprising especially in light of the capsaicin-associated blockade, however, it is consistent with several other immune system related models of pain. PMID- 11879798 TI - Mouse cerebral prostaglandins, but not oxidative damage, change with age and are responsive to indomethacin treatment. AB - Epidemiological and clinical trial data indicate that at least some non-steroidal anti-inflammatory drugs (NSAIDs) reduce the risk of developing Alzheimer's disease (AD). Advancing age is the most robust risk factor for AD. If NSAIDs mitigate the initiation of AD by affecting processes of aging, and if the target of NSAIDs are cyclooxygenases (COX), then COX activity would be hypothesized to increase with advancing age in brain regions affected by AD. We tested this hypothesis in mouse cerebrum by measuring two outcomes of increased COX activity, prostaglandin (PG) levels and markers of oxidative damage. Our results showed that frontal cortical PGE(2) and 6-keto-PGF(1alpha) levels were significantly increased at 14 months compared to 2 months, but that frontal cortical levels of these PGs at 26 months returned to levels observed at 2 months of age. At all ages, 2-week treatment with indomethacin (14 microg/ml drinking water, or approximately 2.2 mg/kg per day) equally suppressed frontal cortical levels of both PGs. In contrast, basal levels of oxidative damage to cerebral cortex did not increase in mice aged up to 26 months, and indomethacin treatment did not significantly alter basal levels of oxidative damage as assayed by F(2) isoprostanes or protein carbonyls. These results suggest that indomethacin may influence the initiation of AD by reducing cerebral PG elevation that may occur in middle age, but that it is unlikely to have a direct effect on levels of oxidative damage. PMID- 11879799 TI - Variability analyses suggest that supraspino-spinal interactions provide dynamic stability in motor control. AB - Effects of supraspino-spinal feedforward-feedback (FF-FB) interactions on variability in locomotor rhythm and coordination were examined in in vitro brain spinal cord lamprey preparations. Spinal locomotor networks were activated by applying 0.2 mM N-methyl-DL-aspartate (NMA) to three spinal pools: gill, rostral and caudal. Bathing the brain with zero Ca(2+) saline altered supraspinal-spinal drive and FF-FB interaction while spino-supraspinal feedback was changed by applying NMA to the caudal pool only. Wavelet analyses indicated a non-uniform energy distribution in ventral root (VR) activity that shifted between frequency bands on FF-FB interruption. Wavelet analysis was used to extract 300-s long epochs of low frequency burst rhythm. These were analyzed using a sliding-window time-varying covariance method. From the autocovariance in each window, the cycle period and height of the first side lobe peak were determined. Rostral VR variability (determined from standard deviation and coefficient of variation of all cycle periods and the mean peak height) was significantly higher than caudal VR variability. FF-FB interruption significantly decreased the rostral VR cycle period and variability but the rostro-caudal gradient remained. The intersegmental delay was also affected. The caudal VR rhythm with NMA in the caudal pool only was slower but more variable than with NMA over the entire cord. These results indicate that the locomotor rhythm in the presence of supraspino spinal interactions is slower but has a higher variability. The higher variability may reflect a dynamic stability of the system. Additionally, differences in local neural organization likely contribute to rostro-caudal differences in variability of the motor output. PMID- 11879800 TI - Aluminum citrate uptake by immortalized brain endothelial cells: implications for its blood-brain barrier transport. AB - The objective was to further test the hypothesis that aluminum (Al) citrate transport across the blood-brain barrier is mediated by a monocarboxylate transporter (MCT). Speciation calculations showed that Al citrates were the predominant Al species under the conditions employed. Al citrate did not inhibit lactate uptake and was not taken up by the rat erythrocyte, suggesting it does not serve as an effective substrate for either MCT1 or the anion exchanger. Studies were conducted with b.End5 cells derived from mouse brain endothelial cells to identify the properties of the carrier(s) mediating Al citrate transport. Western blot analysis of b.End5 cells showed expression of the transferrin receptor and MCT1, but not MCT2 or MCT4. Uptake of Al citrate was approximately 70% faster than citrate. Citrate and Al citrate uptake were sodium independent. Citrate uptake increased at pH 6.9 compared to 7.4, whereas Al citrate uptake did not. Al citrate uptake was reduced by inhibitors of mitochondrial respiration and oxidative phosphorylation, suggesting ATP dependence, but not by ouabain, suggesting no role for Na/K-ATPase. Uptake was not affected by alpha-ketoglutarate or malonate, substrates for the dicarboxylate carrier. Many substrates and inhibitors of MCT1 and organic anion transporters reduced Al citrate uptake into b.End5 cells. BSP and fluorescein, organic anion transporter substrates/inhibitors, inhibited Al citrate uptake. We conclude that Al citrate transport across the blood-brain barrier is carrier-mediated, involving either an uncharacterized MCT isoform expressed in the brain such as MCT7 or MCT8 and/or one of the many members of the organic anion transporting protein family, some of which are known to be expressed at the blood-brain barrier. PMID- 11879801 TI - Selectivity of the protective effects of dihydropyridine calcium channel antagonists against the ethanol withdrawal syndrome. AB - Four dihydropyridine calcium channel antagonists were compared for their ability to protect against the hyperexcitability produced in mice by withdrawal from chronic ethanol treatment and to protect against seizures due to bicuculline or pentylenetetrazol. Comparison was also made of their effects on locomotor activity, body temperature and motor co-ordination, and with the corresponding effects of the benzodiazepine, diazepam. Nitrendipine, nimodipine, nicardipine (at 50 and 10 mg/kg) and isradipine (at 10 and 4 mg/kg) decreased the withdrawal hyperexcitability, but showed no anticonvulsant action against either bicuculline or pentylenetetrazol. Diazepam (1.5 and 4 mg/kg) both protected against the withdrawal signs and decreased seizure incidence after bicuculline and pentylenetetrazol, although the latter effects were of shorter duration than those on the withdrawal signs. The four dihydropyridines decreased spontaneous locomotor activity, an effect which lasted up to 6 h. Only isradipine and diazepam had any ataxic actions at the doses tested. All the dihydropyridines had hypothermic actions, considerably shorter in duration than effects on withdrawal hyperexcitability, with little evidence of dose dependence, except for nicardipine, which had a larger, dose-related, hypothermic action. Of the four compounds, isradipine was more potent in terms of dose, but not any more selective for effectiveness against the withdrawal signs, than the other three dihydropyridines, and nicardipine was slightly less effective in protecting against the withdrawal signs. The results indicate that the anticonvulsant effects of the dihydropyridines were selective for ethanol withdrawal hyperexcitability, whereas diazepam showed no such selectivity. PMID- 11879802 TI - Removal of Ca(2+) following depolarization-evoked cytoplasmic Ca(2+) transients in freshly dissociated pyramidal neurones of the rat dorsal cochlear nucleus. AB - Cytoplasmic [Ca(2+)] ([Ca(2+)](i)) was measured using Fura-2 in pyramidal neurones isolated from the rat dorsal cochlear nucleus (DCN). The kinetic properties of Ca(2+) removal following K(+) depolarization-induced Ca(2+) transients were characterized by fitting exponential functions to the decay phase. The removal after small transients (<82 nM peak [Ca(2+)](i)) had monophasic time course (time constant of 6.43 +/- 0.48 s). In the cases of higher Ca(2+) transients biphasic decay was found. The early time constant decreased (from 3.09 +/- 0.26 to 1.46 +/- 0.11 s) as the peak intracellular [Ca(2+)] increased. The value of the late time constant was 18.15 +/- 1.60 s at the smallest transients, and showed less dependence on [Ca(2+)](i). Blockers of Ca(2+) uptake into intracellular stores (thapsigargin and cyclopiazonic acid) decreased the amplitude of the Ca(2+) transients and slowed their decay. La(3+) (3 mM) applied extracellularly during the declining phase dramatically changed the time course of the Ca(2+) transients as a plateau developed and persisted until the La(3+) was present. When the other Ca(2+) removal mechanisms were available, reduction of the external [Na(+)] to inhibit the Na(+)/Ca(2+) exchange resulted in a moderate increase of the time constants. It is concluded that in the isolated pyramidal neurones of the DCN the removal of Ca(2+) depends mainly on the activity of Ca(2+) pump mechanisms. PMID- 11879803 TI - Behavioral activity and stereotypy in rats induced by L-DOPA metabolites: a possible role in the adverse effects of chronic L-DOPA treatment of Parkinson's disease. AB - L-3,4-Dihydroxyphenylalanine (L-DOPA) is a common and effective treatment for Parkinson's disease, but dyskinesia continues to be a serious adverse effect with chronic use. Evidence suggests that L-DOPA induces increases in dopamine, which then binds to supersensitive dopamine receptors, resulting in dyskinesia. We have shown previously that L-DOPA directly causes stereotypy in rats, suggesting that chronic L-DOPA-induced dyskinesia is also caused by L-DOPA itself. This raises the possibility that other L-DOPA metabolites have a role in dyskinesia. We examined the behavioral effects of five L-DOPA metabolites (3-methoxytyramine, 3 MT; 3,4-dihydroxyphenylalanine, DOPAC; dopamine; homovanillic acid, and 3-o methyl-DOPA) in rats. A unilateral, intracerebroventricular injection of 3-MT (10 200 microg, 40 microl) over 30 min, dose-dependently increased behavioral activity and stereotypy. This effect was suppressed by the dopamine D1/5-receptor antagonist SCH 23390, but not by the dopamine D2/3/4-receptor antagonist sulpiride. Dopamine denervation resulted in behavioral supersensitivity to 3-MT. Neither dopamine nor DOPAC levels increased in the striatum after 3-MT administration, as measured using in vivo voltammetry. The behavioral changes paralleled a rise in 3-MT in the contralateral striatum. DOPAC also caused behavioral changes and stereotypy, but to a smaller degree than 3-MT. Dopamine denervated rats did not exhibit a supersensitive response to DOPAC, however. Other L-DOPA metabolites did not cause behavioral effects. These data suggest that 3-MT directly induced dopamine-D1/5-receptor-mediated behavioral changes in rats, and that 3-MT may have a role in dyskinesia due to chronic L-DOPA treatment in Parkinson's disease patients. PMID- 11879804 TI - Characterization of parkin in bovine peripheral nerve. AB - The autosomal recessive juvenile parkinsonism is caused by the mutations of the gene encoding a novel protein called parkin. It has been reported that parkin is expressed in the central nervous system and functions as a ubiquitin-protein ligase (E3) which suppresses neuronal cell degeneration by ubiquitinating misfolded proteins. Thus far, however, it remains unknown if parkin is expressed and functions in the peripheral nervous system. In order to begin to address to this question, we investigated the expression of parkin in bovine peripheral nerve. Reverse transcription polymerase chain reaction analysis demonstrated the presence of parkin transcript in bovine peripheral nerve. The obtained bovine parkin cDNA sequence was identical to that of human except a single nucleotide. Immunoblot analysis demonstrated the expression of parkin protein in bovine peripheral nerve. Immunohistochemical analysis demonstrated the localization of parkin in the axoplasm of myelinated nerve fibers, the Schwann cell cytoplasm and the Schwann cell outer membrane. Furthermore, fractionation analysis indicated the presence of two fractions of parkin in bovine peripheral nerve, the cytosolic fraction and the cell membrane-bound fraction. All together, these results point to diverse roles of parkin in not only the central but also the peripheral nervous system. PMID- 11879805 TI - Mu and delta opioid receptor-like immunoreactivity in the cervical spinal cord of the rat after dorsal rhizotomy or neonatal capsaicin: an analysis of pre- and postsynaptic receptor distributions. AB - Opioid compounds have powerful analgesic properties when administered to the spinal cord. These effects are exerted through mu and delta opioid receptors, and both pre- and postsynaptic mechanisms have been implicated. To specifically address the relative pre- and postsynaptic contribution to spinal opioid analgesia, we have quantitatively assessed the pre- vs. postsynaptic distribution of the mu-opioid (MOR-1, MOP(1)) and delta-opioid receptors (DOR-1, DOP(1)). We also examined the rostro-caudal arborization of MOR-1 and DOR-1 immunoreactive primary sensory neurons, using an isolated dorsal root preparation. These results were compared to those obtained by labeling for calcitonin gene-related peptide (CGRP), a neuropeptide whose expression in the spinal cord is restricted to the terminals of small diameter primary sensory neurons. We estimate that approximately one half of MOR-1 and two thirds of DOR-1 immunoreactivity in the cervical spinal cord is located on primary afferent fibers. These fibers have a broad rostro-caudal distribution, extending at least three segments rostral and caudal to their segment of entry. Regardless of marker used, the rostral projection was greatest, however, the distribution of CGRP-immunoreactive fibers differed somewhat in that they had a much smaller projection to the most caudal segments examined. Our results suggest that presynaptic delta opioid actions predominate, but that there are mixed pre- and postsynaptic inhibitory effects exerted by opioid analgesics that act at the spinal cord mu opioid receptor. PMID- 11879806 TI - Antisense oligonucleotides to N-methyl-D-aspartate receptor subunits attenuate formalin-induced nociception in the rat. AB - Noxious peripheral stimuli increase the sensitivity of central nociceptive neurons to subsequent noxious stimuli. This occurs in part through activation of spinal N-methyl-D-aspartate (NMDA) receptors. These receptors are heteromeric complexes of NMDA-R1 and NMDA-R2 A--D subunits. NMDA-R1 is necessary for the formation of functional NMDA receptors whereas the R2 subunits (A-D) modify the properties of the receptor. However, the role of the various receptor subtypes in nociception has not been established. In this study, we used intrathecally administered phosphodiester antisense oligonucleotides (ODEs) to examine the role of the NMDA-R1, NMDA-R2C and NMDA-R2D subunits in the mediation of formalin induced nociception in the rat. The antisense ODEs against the NMDA-R1 and NMDAR 2C subunits reduced nociceptive behaviors whereas the corresponding sense ODEs had no effect. In contrast, nociception was unaffected by the antisense ODE to NMDAR-2D. Using an RNase protection assay, we also found that each antisense ODE selectively decreased the level of the corresponding mRNA in the lumbar spinal cord but that the sense ODEs had no such effect. Accordingly, these data provide evidence that the R1 and R2C subunits, but not R2D, of the NMDA receptor participate in the development of formalin-induced nociception. PMID- 11879807 TI - Expression and activity of antioxidants in the brain in progressive supranuclear palsy. AB - Recent evidence implicates oxidative stress in the pathophysiology of progressive supranuclear palsy (PSP). Thus, we undertook a study of the activity and localization of two essential antioxidant systems (superoxide dismutase [SOD] enzymes and total glutathione) in the human post-mortem PSP and control brain. Marked increases in SOD1 (Cu/ZnSOD) activity and glutathione levels were measured within most PSP brain regions examined, whereas, only the subthalamic nucleus exhibited a significant increase (+68%) in SOD2 (MnSOD) activity. Two additional cases with mild pathological abnormalities were studied. The first (case A) may represent an example of an asymptomatic PSP case, while the second (case B) had mild pathological abnormalities consistent with typical PSP. In case A, only the STN had elevated levels of SOD activity, in the absence of an increase in tissue glutathione content. In case B, SOD activities and tissue glutathione content were elevated in several regions. Immunolocalization of the SOD1 and SOD2 proteins in paraffin-embedded tissue sections revealed a marked increase in the density of SOD immunopositive profiles (particularly glia) in the typical PSP brain, particularly within the white matter. Together, our data argues strongly in favor of the involvement of oxidative stress in the etiology and progression of PSP, and suggests that deficit in SOD or glutathione metabolism are not causative. PMID- 11879808 TI - Influence of gender on dilatation of the basilar artery during diabetes mellitus. AB - The first goal of the present study was to examine the influence of gender on reactivity of the basilar artery. The second goal of this study was to examine the effect of diabetes mellitus on reactivity of the basilar artery in male and female rats. We examined in vivo responses of the basilar artery in male and female nondiabetic and diabetic rats in response to a nitric oxide synthase (NOS) dependent (acetylcholine) and -independent (nitroglycerin) agonist. In nondiabetic male and female rats, acetylcholine and nitroglycerin produced dose related dilatation of the basilar artery. However, the magnitude of vasodilatation in response to acetylcholine and a high concentration of nitroglycerin was significantly greater in female than in male rats. Acetylcholine (1.0 microM) dilated the basilar artery by 11 +/- 2% in nondiabetic males versus 25 +/- 4% in nondiabetic females (P<0.05). Nitroglycerin (1.0 microM) dilated the basilar artery by 37 +/- 8% in nondiabetic males versus 62 +/ 5% in nondiabetic females (P<0.05). Thus, there is a significant effect of gender on reactivity of the basilar artery during physiologic conditions. Dilatation of the basilar artery in response to acetylcholine, but not nitroglycerin, was impaired in diabetic male and female rats compared to their nondiabetic counterparts. Acetylcholine (1.0 microM) dilated the basilar artery by only 5 +/- 1% in diabetic males and by only 4 +/- 1% in diabetic females. In summary, dilatation of the basilar artery in response to NOS-dependent agonist was significantly greater in nondiabetic female than in nondiabetic male rats. In addition, diabetes mellitus impaired NOS-dependent dilatation of the basilar artery not only in male rats, but also in female rats. We suggest that the results of these studies provide insight into the pathogenesis of cerebrovascular abnormalities observed in postmenopausal women. PMID- 11879809 TI - Expression changes of somatostatin receptor subtypes sst2A, sst2B, sst3 and sst4 after a cortical contusion trauma in rats. AB - The neuropeptide somatostatin acts as a neuromodulator in the CNS in a predominantly inhibitory manner. In this study, an ipsilateral cortical and hippocampal damage in the brain of adult rats was induced by a cortical contusion trauma in order to examine subsequent changes of expression of different somatostatin receptor subtypes (sst). By using subtype specific antibodies we found a clear decline of expression level for sst2A, sst2B, sst3 and sst4 subtypes in the pyramidal cell layer of the ipsilateral hippocampus. Nissl staining revealed that this decline of expression level is due to cell death of sst expressing neurons within the first 48 h after trauma. Additionally we found a progressive infiltration of sst4 positive cells into regions of cortical and hippocampal damage. The number of these cells increases strikingly within the first 3 days after trauma and it seems that their morphology changes from a round to an astrocyte-like shape. Moreover, sst4 and sst2A positive cells accumulate in the ipsilateral ependym and pyramidal-like cells expressing sst4 were found beneath the damaged CA3 pyramidal layer. Taken together, after trauma we found deterioration of sst positive neurons and an additional activation of sst4 and sst2A expressing cells the final fate of which has to be elucidated further. PMID- 11879810 TI - Ischemic preconditioning reduces infarct volume after subdural hematoma in the rat. AB - The aim of the present study was to investigate the effects of ischemic preconditioning on infarct volume in a rat model of subdural hematoma (SDH). Ischemic preconditioning was induced by 15 min transient middle cerebral artery (MCA) occlusion followed 3 days later by the injection of 300 microl of autologous venous blood into the subdural space. Preconditioning significantly reduced the volume of cortical infarction (by 26%, P<0.001) 24 h after SDH induction, but not brain swelling (P>0.05) relative to sham-operated non preconditioned animals. These data support the view that ischemic preconditioning reduces ischemic brain damage in this rat model of SDH. PMID- 11879812 TI - Electroencephalogram theta frequency changes in parallel with euthermic brain temperature. AB - To test whether moderate changes in brain temperature can influence electroencephalogram (EEG) frequencies in a significant way the behaviour of peak theta frequency (approximately 6 Hz) in the rapid-eye movement (REM) sleep EEG was investigated in the Djungarian hamster under influence of the spontaneous euthermic changes in brain temperature. In contrast to in vitro data, collected from hippocampal tissue, theta peak frequency shifted significantly when temperature changed between 33 and 37 degrees C. The results support the hypothesis that moderate changes in brain temperature can influence EEG power density spectra. PMID- 11879811 TI - Age-related changes in hypocretin (orexin) immunoreactivity in the cat brainstem. AB - Terminals of hypothalamic hypocretin-containing neurons are observed within brainstem nuclei involved in the control of sleep and wakefulness. Because aged humans, cats and other species exhibit changes in sleep and wakefulness in old age, we were interested in examining age-related changes in hypocretin/orexin projections to the following brainstem regions which are associated with the regulation of sleep and wakefulness: the dorsal raphe nucleus, the laterodorsal tegmental nucleus, the pedunculo-pontine tegmental nucleus and the locus coeruleus. Based upon the results of immunohistochemical determinations, in all the regions examined, round or oval "spot-like" structures were observed in aged cats. Many of these "spot-like" structures resembled enlarged varicosities of a nature that would be expected to disrupt hypocretin neurotransmission. In addition, a site-specific decrease in immunostaining was observed in the locus coeruleus in old cats compared with adult controls; this result likely reflects a decrease in the number of labeled fibers, which indicates that there occurs a degeneration of hypocretinergic function in conjunction with old age. The proceeding changes may account for some of sleep-wake disturbance which are observed in aged animals as well as elderly humans. PMID- 11879813 TI - Dual action of estrogen on glutamate-induced calcium signaling: mechanisms requiring interaction between estrogen receptors and src/mitogen activated protein kinase pathway. AB - Conjugated equine estrogens (CEE) is the most widely prescribed pharmaceutical estrogen replacement therapy (ERT) for postmenopausal women in the United States and is the ERT of the Women's Health Initiative. Previous studies from our laboratory have demonstrated that CEE exerts neurotrophic and neuroprotective effects in neurons involved in learning and memory, and which are affected in Alzheimer's disease. The present work demonstrates that CEE potentiated the rise in intracellular calcium ([Ca(2+)](i)) following exposure to physiological concentrations of glutamate. In contrast, the reverse effect occurred in the presence of excitotoxic levels of glutamate exposure, where CEE attenuated the rise in [Ca(2+)](i). Potentiation of the glutamate response was mediated by the NMDA receptor, as the NMDA receptor antagonist MK-801 blocked the CEE-induced potentiation, whereas the L-type calcium channel blocker nifedipine did not. Further, the CEE-potentiated glutamate response was mediated by a src tyrosine kinase, as the tyrosine kinase inhibitor PP2 blocked the potentiation induced by CEE and neurons treated with CEE displayed increased phosphorylated tyrosine. The inhibition by CEE of [Ca(2+)](i) rise in the presence of excitotoxic levels of glutamate was mediated by mitogen activated protein kinase (MAPK), as the protective effect of CEE was blocked by inhibiting MAPK activation with PD98059. These data provide potential mechanisms to explain the cognitive enhancing and neuroprotective effects exerted by ERT. PMID- 11879814 TI - In vivo assessment of nodularin-induced hepatotoxicity in the rat using magnetic resonance techniques (MRI, MRS and EPR oximetry). AB - Acute nodularin-induced hepatotoxicity was assessed in vivo, in rats using magnetic resonance (MR) techniques, including MR imaging (MRI), MR spectroscopy (MRS), and electron paramagnetic resonance (EPR) oximetry. Nodularin is a cyclic hepatotoxin isolated from the cyanobacterium Nodularia spumigena. Three hours following the intraperitoneal (i.p.) administration of nodularin (LD50), a region of 'damage', characterized by an increase in signal intensity, was observed proximal to the porta hepatis (PH) region in T2-weighted MR images of rat liver. Image analysis of these regions of apparent 'damage' indicated a statistically significant increase in signal intensity around the PH region following nodularin administration, in comparison with controls and regions peripheral to the PH region. An increase in signal intensity was also observed proximal to the PH region in water chemical shift selective images (CSSI) of nodularin-treated rat livers, indicating that the increased signal observed by MRI is an oedematous response to the toxin. Microscopic assessment (histology and electron microscopy) and serum liver enzyme function tests (aminotransferase (ALT) and aspartate ALT (AST)) confirmed the nodularin-induced tissue injury observed by MRI. In vivo and in vitro MRS was used to detect alterations in metabolites, such as lipids, Glu+Gln, and choline, during the hepatotoxic response (2-3 h post-exposure). Biochemical assessment of perchloric acid extracts of nodularin-treated rat livers were used to confirm the MRS results. In vivo EPR oximetry was used to monitor decreasing hepatic pO2 (approximately 2-fold from controls) 2-3 h following nodularin exposure. In vivo MR techniques (MRI, MRS and EPR oximetry) are able to highlight effects that may not have been evident in single end point studies, and are ideal methods to follow tissue injury progression in longitudinally, increasing the power of a study through repeated measures, and decreasing the number of animals to perform a similar study using histological or biochemical techniques. PMID- 11879815 TI - Comparative quantitative structure toxicity relationships for flavonoids evaluated in isolated rat hepatocytes and HeLa tumor cells. AB - Quantitative structure activity relationship (QSAR) equations were obtained to describe the cytotoxicity of 22 polyphenols using toxicity (logLD50) representing the concentration for 50% cell survival in 2 h for isolated rat hepatocytes, log P representing octanol/water partitioning, and/or E(p/2) representing redox potential. One- and two-parameter equations were derived for the quantitative structure toxicity relationships (QSTR) for polyphenol induced hepatocyte cytotoxicity: e.g. log C(hepatocyte) (microM)=-0.65(-0.08)log P+4.12(-0.15) (n=19, r(2)=0.80, s=0.33, P<1 x 10(-6)). One- and two-parameter QSAR equations were also derived to describe the inhibitory effects of 13 polyphenols on tumor cell growth when incubated with HeLa cells for 3 days: e.g. log C(tumor) (microM)=-0.34(+/-0.04)log P+2.40(+/-0.07) (n=11, r(2)=0.90, s=0.13, P<1 x 10( 5)). These findings point to lipophilicity as a major characteristic determining polyphenol cytotoxicity. The E(p/2) also played a significant role in polyphenol cytotoxicity towards both cell types: e.g. log C(hepatocyte) (microM)=-0.60(+/ 0.06)log P+2.01(+/-0.43)E(p/2) (V)+3.86(+/-0.12) (n=9, r(2)=0.96, s=0.15, P<0.005). The involvement of log P and E(p/2) could be explained if polyphenol cytotoxicity involved the formation of radicals, which interacted with the mitochondrial inner membrane resulting in a disruption of the membrane potential. PMID- 11879816 TI - Cytotoxicity mechanisms of sodium hypochlorite in cultured human dermal fibroblasts and its bactericidal effectiveness. AB - We investigated the therapeutic efficacy of the topical antiseptic sodium hypochlorite (NaOCl) for antibacterial activity and in parallel the cytotoxicity mechanisms by which hypochlorite and the chloramines generated therefrom induce oxidative tissue damage, which further influences the wound-healing process. Human dermal fibroblasts were exposed to increasing concentrations of reagent NaOCl (0.00005-0.1%) at exposure times varying between 2 and 24 h and the protective effects of fetal calf serum (FCS) determined. Antibacterial power was studied by testing a wide range of hypochlorite concentrations (0.00025-0.5%) against four isolated bacterial species. Total bactericidal effects were observed only for 0.5%; concentration range 0.25-0.025% produced partial antimicrobial activity. The early NaOCl-produced cytotoxic action on cultured fibroblasts was cell ATP depletion which occurred at 0.00005% (with FCS 2%) followed by dose- and time-dependent decreases, reaching levels below 5% of control values. Using the 3'-[1-(phenylamino-carbonyl)-3,4-tetrazolium]-bis(4-methoxy-6-nitro)benzene sulfonic acid metabolic assay to evaluate cell death, we observed that NaOCl concentrations greater than 0.05% provoked null fibroblast survival at all exposure times assayed. Hypochlorous acid proved to exert a rapid inhibitory effect on DNA synthesis, consistent with its primary role in bacterial killing by phagocytes. Cytotoxicity produced by increasing NaOCl concentrations and assessed by measuring both mitochondrial function and cell DNA synthesis was reduced with the greatest presence of FCS (10%) in culture media. PMID- 11879819 TI - Modulation of selected cell signaling genes in mouse liver by fumonisin B1. AB - Fumonisin B1 (FB1) is a naturally occurring mycotoxin produced primarily by Fusarium verticillioides and related fungi, common contaminants of corn throughout the world. FB1 is a carcinogen and causative agent of several lethal animal diseases, including equine leukoencephalomalacia and porcine pulmonary edema. Liver is the primary target organ in mice. In vivo and vitro, cells exposed to FB1 undergo a mixture of necrotic and apoptotic cell death. Our previous studies showed gender differences in hepatotoxicity caused after 5 day FB1 treatment. Gene alterations in cytokine network and apoptosis signaling molecules were also observed after an acute single dose of FB1. To further investigate the gene alterations after a subchronic FB1 exposure and its correlation to observed gender differences, male and female BALB/c mice (five per group) were injected subcutaneously with either saline or 2.25 mg/kg per day of FB1 for 5 days. FB1 caused increased expression of tumor necrosis factor alpha (TNFalpha), interleukin (IL)-1alpha, IL-1beta, IL-1 receptor antagonist (IL-1Ra), IL-6, IL-10, IL-12 p40, IL-18 and interferon gamma (IFNgamma) in male liver, with a similar increase in females except for IL-1beta and IL-18. Control females showed higher basal levels of IL-1alpha, IL-1Ra, IL-10, IL-12 p40 and IFNgamma compared with males. Expression of TNF receptor 55 and TNF receptor associated death domain (TRADD) was increased, with no changes in Fas signaling molecules, Fas, Fas ligand (FasL), Fas associated death domain (FADD) and Fas-associated protein factor (FAF). Expression of oncogenic transcription factors, c-Myc, B Myc, Max and Mad, and apoptotic genes, namely Bcl-2, Bax and Bad, was increased after FB1 treatment. FB1 caused an activation of cytokine network in liver, particularly the TNFalpha signaling pathway, suggesting its involvement in hepatotoxic mechanisms. Induction of IL-1Ra and oncogenes is a likely mechanism for the cancer promoting properties of FB1, through a mechanism involving apoptotic necrosis, oncotic necrosis and consequent regeneration. PMID- 11879817 TI - Comparative studies on biological activity of certain microtubule-interacting taxanes. AB - We have compared the nature of interaction of certain taxanes with microtubular protein, and the mechanism of action underlying cytotoxic activity. Taxanes induced tubulin assembly in vitro, but only taxanes bearing side chain were capable of inducing the formation of stable tubulin polymers. Electron microscopy detections showed that taxane-induced polymers are structurally similar to microtubules formed by paclitaxel, with differences in length. Otherwise, light microscopy views have shown that intracellular microtubule network is deeply reorganized by taxanes into short fibers, unlike paclitaxel-bundled microtubules. Taxanes inhibited the growth of various human tumor cell lines, but cell cycle analysis did not always indicate a block in the G2/M phase. These agents alter some apoptotic signal transduction pathways, probably by a mechanism distinct from microtubule interaction. Briefly, the effectiveness of taxanes is closely related to their chemical structure, and depends on their interaction with microtubular protein. By virtue of this mechanism, some of these taxanes may provide usefulness for therapeutic improvements. PMID- 11879818 TI - Sex hormone-regulated renal transport of perfluorooctanoic acid. AB - The biological half-life (t1/2) of perfluorooctanoic acid (PFOA) in male rats is 70 times longer than that in female rats. The difference is mainly due to the difference in renal clearance (CL(R)), which was significantly reduced by probenecid, suggesting that PFOA is excreted by organic anion transporter(s). Castration of male rats caused a 14-fold increase in the CL(R) of PFOA, which made it comparable with that of female rats. The elevated PFOA CL(R) in castrated males was reduced by treating them with testosterone. Treatment of male rats with estradiol increased the CL(R) of PFOA. In female rats, ovariectomy caused a significant increase in CL(R) of PFOA, which was reduced by estradiol treatment. Treatments of female rats with testosterone reduced the CL(R) of PFOA as observed in castrated male rats. To identify the transporter molecules that are responsible for PFOA transport in rat kidney, renal mRNA levels of organic anion transporter 1 (OAT1), OAT2, OAT3, organic anion transporting polypeptide 1 (oatp1), oatp2 and kidney specific organic anion transporter (OAT-K) were determined in male and female rats under various hormonal states and compared with the CL(R) of PFOA. The level of OAT2 mRNA in male rats was only 13% that in female rats. Castration or estradiol treatment increased the level of OAT2 mRNA whereas treatment of castrated male rats with testosterone reduced it. In contrast to OAT2, mRNA levels of both oatp1 and OAT-K were significantly higher in male rats compared with female rats. Castration or estradiol treatment caused a reduction in the levels of mRNA of oatp1 and OAT-K in male rats. Ovariectomy of female rats significantly increased the level of OAT3 mRNA. Multiple regression analysis suggests that the change in the CL(R) of PFOA is, at least in part, due to altered expression of OAT2 and OAT3. PMID- 11879820 TI - Multi-generational analysis and visualization of the vascular tree in 3D micro-CT images. AB - Micro-CT scanners can generate large high-resolution three-dimensional (3D) digital images of small-animal organs, such as rat hearts. Such images enable studies of basic physiologic questions on coronary branching geometry and fluid transport. Performing such an analysis requires three steps: (1) extract the arterial tree from the image; (2) compute quantitative geometric data from the extracted tree; and (3) perform a numerical analysis of the computed data. Because a typical coronary arterial tree consists of hundreds of branches and many generations, it is impractical to perform such an integrated study manually. An automatic method exists for performing step (1), extracting the tree, but little effort has been made on the other two steps. We propose an environment for performing a complete study. Quantitative measures for arterial-lumen cross sectional area, inter-branch segment length, branch surface area and others at the generation, inter-branch, and intra-branch levels are computed. A human user can then work with the quantitative data in an interactive visualization system. The system provides various forms of viewing and permits interactive tree editing for "on the fly" correction of the quantitative data. We illustrate the methodology for 3D micro-CT rat heart images. PMID- 11879821 TI - An approach to computer automation of the extracorporeal circulation. AB - In order to move towards extracorporeal circulation (ECC) automation, a virtual simulation of the process was designed. The ECC model is composed of a virtual patient linked to a virtual ECC circuit. A user interface panel allows to set control parameters for the simulation and to visualize results. It is possible to switch between manual and automatic control. Meaningful hemodynamic and hematochemical variables are continuously shown along with a score (from 0 to 10). The virtual model can play a crucial role in educating and training the personnel devoted to the managing of the heart-lung machine. PMID- 11879822 TI - Time-critical dynamic decision modeling in medicine. AB - Many real-world medical applications require timely actions to be taken in time pressured situations. Existing approaches to dynamic decision modeling have provided relatively efficient methods for representing and reasoning, but the process of computing the optimal solution has remained intractable. A major reason for this difficulty is the lack of models that are capable of modeling temporal processes and dealing with time-critical situations. This paper presents a formalism called the time-critical dynamic influence diagram that provide the capability for both temporal and space abstraction. To deal with the time criticality, we exploit the concept of space and temporal abstraction to reduce the computational complexity and propose an anytime algorithm for the solution process. We illustrate through out the paper, the various approaches with the use of a medical problem on the treatment of cardiac arrest. PMID- 11879823 TI - Perceptron error surface analysis: a case study in breast cancer diagnosis. AB - Perceptrons are typically trained to minimize mean square error (MSE). In computer-aided diagnosis (CAD), model performance is usually evaluated according to other more clinically relevant measures. The purpose of this study was to investigate the relationship between MSE and the area (A(z)) under the receiver operating characteristic (ROC) curve and the high-sensitivity partial ROC area ((0.90)A'(z)). A perceptron was used to predict lesion malignancy based on two mammographic findings and patient age. For each performance measure, the error surface in weight space was visualized. Comparison of the surfaces indicated that minimizing MSE tended to maximize A(z), but not (0.90)A'(z). PMID- 11879824 TI - Predictors of outcome following hip fracture. Admission time predicts length of stay and in-hospital mortality. AB - Many factors may contribute to the mortality and morbidity following hip fracture, including the provision of care. We wished to examine the contribution of potential factors to in-hospital mortality, length of hospital stay and 90-day mortality by statistical analyses of an audit database of all hip fractures admitted to a teaching hospital following the introduction of a fast track admission system. In-hospital mortality was predicted by ASA grade, the presence of any complications, cardiovascular complications, grade of surgeon, operation type and shorter admission time, a measure of time taken to admit a patient to a hospital bed (P<0.001). Length of hospital stay was predicted by increased age, presence of chronic cognitive impairment/dementia, presence of an implant complication, operation type, fracture type and longer admission time, r=0.455, P<0.001. Ninety-day mortality was predicted by the presence of chronic cognitive impairment/dementia, cardiovascular complications, pulmonary complications, ASA grade, grade of surgeon and admission day, P<0.001. Rapid admission following a hip fracture may not be the ideal management approach for all patient groups. Further study is required to identify factors in the process of care which are associated with better outcomes. PMID- 11879825 TI - Hip fracture rehabilitation -- a comparison of two centres. AB - The outcomes for 2005 consecutive patients aged 50 years or over admitted to two orthopaedic centres with a hip fracture were prospectively studied. All the patients were followed up to 1 year from injury. The mean orthopaedic ward stay was shorter in Edinburgh in comparison to Peterborough (20 versus 10 days), but the total hospital stay was shorter in Peterborough (34 versus 22 days). In Peterborough twice as many patients were discharged directly back from the orthopaedic ward to their place or origin (82 versus 41%). At 1 year from injury 34% of the patients had died. Factors associated with an increased mortality were increased age, male sex, pre-fracture place of residence and impaired mobility. For the survivors, the patients from Edinburgh had an increased mortality but were older, more likely to have impaired mobility and to come from more dependent residential accommodation. The different mortality between centres (30 versus 36%) was not statistically significant when adjusted for the possible confounding factors. PMID- 11879827 TI - Failure of Austin Moore hemiarthroplasty: total hip replacement as a treatment strategy. AB - Forty six patients who underwent conversion of their Austin Moore hemiarthroplasty to a total hip replacement were assessed clinically and radiologically. Forty patients requiring conversion had either groin or thigh pain. Operative findings showed acetabular cartilage degeneration and femoral loosening as the main pathological process causing their symptoms. Total hip replacement gave complete resolution of the symptoms in 41 patients. We propose early conversion to total joint arthroplasty in patients with painful hemiarthroplasties. PMID- 11879826 TI - Cemented or uncemented hemiarthroplasty for displaced intracapsular fractures of the hip--a systematic review. AB - Displaced intracapsular hip fractures are often treated by hemiarthroplasty, with or without cement. The aim of this study is to perform a systematic review of the literature to establish whether this influences the outcome. A comprehensive search of the English language world literature of all studies comparing cemented with uncemented hemiarthroplasty was performed. Eighteen publications addressing this issue were identified. Three were excluded from review because of poor study design and non-comparability of groups. Of the remainder, few were suitable for statistical analysis, therefore a qualitative comparison of results was carried out. The majority of studies suggested a lower revision rate, less thigh pain and better mobility in-patients in whom the prosthesis was cemented. Cemented hemiarthroplasty may be associated with increased operative time and blood loss. There appears to be no difference in general complication, or mortality rates after 3 months, between the two groups. Radiographic differences were variable and did not correlate with clinical findings. We conclude, the literature tends to support the use of cement in hemiarthroplasty for displaced intracapsular femoral neck fractures. Further prospective randomised studies are required to resolve the issue. PMID- 11879828 TI - Intramedullary hip screw versus sliding hip screw for unstable intertrochanteric femoral fractures in the elderly. AB - We report a randomised, prospective study comparing a standard sliding hip screw and the intramedullary hip screw for the treatment of unstable intertrochanteric fractures in the elderly. One hundred and two patients were randomised on admission to two treatment groups. Fifty-two patients were treated with a compression hip screw (CHS), and fifty had intramedullary fixation with an intramedullary hip screw (IMHS). Patients were followed for 1 year and had a clinical and radiological review at 3, 6 and 12 months. The mean duration of operation and fluoroscopy screening time was significantly greater for insertion of the intramedullary hip screw. There was no difference between the groups with regard to transfusion requirements or time to mobilise after surgery. There were two technical complications in the CHS group and three in the IMHS group. There was no significant difference between the two groups in radiological or functional outcome at 12 months. It remains to be shown whether the theoretical advantages of intramedullary fixation of extracapsular hip fractures bring a significant improvement in eventual outcome. PMID- 11879830 TI - Altered venous function and deep venous thrombosis following proximal femoral fracture. AB - The effect of surgery for femoral neck fracture on lower limb venous blood flow and its relationship to deep vein thrombosis was investigated in 179 patients. Blood flow was measured using strain gauge plethysmography before surgery, in the 1st week after surgery, and at 6 week review. There was a significant reduction in both venous outflow and venous capacitance, affecting both fractured and non fractured legs but significantly greater in the fractured leg. Venous function remained significantly impaired in both lower limbs 6 weeks after surgery. There was a significant correlation between the reduction in venous function and the development of deep vein thrombosis. PMID- 11879829 TI - Simultaneous hip and upper limb fracture in the elderly: incidence, features and management considerations. AB - We performed a retrospective analysis of all patients admitted to a single unit over 4 years with fractured neck of femur. Of the 760 patients admitted over this period of time 36 (4.7%) were found to also have a fracture of the upper limb. The associated upper limb fractures were distal radius (n=28), olecranon (n=2) and neck of humerus (n=5) and fifth metacarpal (n=1), with the same ratio of intracapsular to extracapsular fractures as the whole group. The female to male ratio in the isolated hip fracture group was 2.4:1 and for the combined fractures group was 8:1 (difference P=0.014). The mean patient age was 77.3 years for isolated hip fractures and 83.9 for the combined group (P=0.037). The mean total length of stay in hospital for isolated hip fracture was 15.6 days and for combined fractures was 20.4 days (P=0.010). We have demonstrated that combined upper limb and neck of femur fractures occur in a population that is older and predominantly female. They are associated with a significant increase in hospital stay and increased difficulties in mobilisation as a consequence of the combined fractures. It is therefore important to recognise this specific subgroup of patients presenting with hip fractures in order to ensure that they receive adequate treatment and rehabilitation and that the use of valuable health care resources are optimised. PMID- 11879831 TI - Fractures of the proximal part of the femur in patients under 50 years of age. AB - We present a retrospective study of the epidemiology and the results of treatment of fractures of the proximal femur in 74 patients aged 50 or under. Cervical fractures comprised 17% of the 74 fractures and were equally caused by high and low energy trauma. Trochanteric and subtrochanteric fractures comprised 83% of the total and were mainly caused by high-energy trauma. The rate of complications was 25% for the cervical fractures (two avascular necroses and one non-union) and was attributed to a valgus malalignment and to the use of bulky implants. Similarly the rate of complications was 22.5% for the trochanteric and subtrochanteric fractures (mal-union and displacement into varus) and was attributed to early collapse of the fracture and the inability of the implant to withstand the strain before osseous union. Furthermore, intramedullary fixation of subtrochanteric fractures should be preferred to the sliding screw and side plate. In conclusion, we believe that proximal femoral fractures in young patients require great attention to the correct reduction and the appropriate choice of implant, in order to minimize the chances of the development of serious complications. PMID- 11879832 TI - The Mennen femoral plate for fixation of periprosthetic femoral fractures following hip arthroplasty. AB - Periprosthetic fractures can be treated by various methods. The Mennen femoral plate used to be a common implant in our region to stabilise periprosthetic femoral fractures following hip arthroplasty. This device has been used in 16 patients in our region from three different centres. The periprosthetic femoral fractures occurred approximately 7 years after the hip arthroplasty procedure. After stabilisation of the fracture, patients were kept non-weight bearing. Weight bearing was commenced once there was clinical and radiological evidence of fracture union. About 75% of our patients had a complication following fixation of the fracture. In all of these patients the main complication was varus mal union of the fracture. As per our study the Mennen femoral plate seems to be a weak fixation device. The plate is unable to counter the medial compressive forces on the femur leading to a varus collapse of the fracture. PMID- 11879833 TI - Safety to drive after common limb fractures. AB - One hundred Orthopaedic Consultants in the UK were randomly selected and asked to complete a postal questionnaire indicating when they felt patients with treated, pain free, common limb fractures could return to driving. Seventy-two questionnaires were returned. Six were not completed leaving 66 completed questionnaires. Analysis revealed a clear mandate to allow or prevent driving in patients with certain treatment modalities and stages of union. In the lower limb there was majority agreement of suitability to drive in 61% of the 28 presented scenarios. Opinion was more divided for the upper limb, with only 43% of the scenarios having majority agreement. Our results show that while for some common limb fractures there is a clear body of opinion supporting safety to drive, for other common fractures opinion is less clear cut. This has important financial, medico-legal and legal implications that could be resolved by formal testing. PMID- 11879834 TI - Doctor, can I drive with this plaster? An evidence based response. AB - The purpose of this study is to assess the effect of the commonly used below elbow plaster casts on driving ability. The position of the Driver and Vehicle Licensing Agency and five motor insurance companies is established. The study aims to help doctors decide whether or not a patient is fit to drive with a plaster cast. Three types of cast were tested using one driver. A score was given for several driving abilities. The right Colles cast was found to have no effect on ability to drive. Scaphoid and Bennett's casts were found to have significant affects on driving ability. The DVLA has no specific guidelines regarding driving with a plaster cast and the position of insurance companies is variable, but will usually depend upon medical advice. PMID- 11879835 TI - Upper root brachial plexus trauma; patient selection and reconstruction. AB - Injury to the brachial plexus is increasingly common and the initial management of these patients is usually focused on associated life threatening injuries. Appreciation of the management of the brachial plexus injury can greatly assist with subsequent reconstruction and thus we review our experience in this field. A total of nine patients who underwent upper root brachial plexus reconstruction over the period 1980-1998 were reviewed. Causes of injury included road traffic accidents (n=6), open injuries (n=2) and the remaining case was iatrogenic. All patients had cabled grafting of the plexus while one patient had neurotization of the plexus in addition to grafting. Sixty six percent (n=6/9) of patients had a good outcome with return of elbow flexion. Patients with an open injury to the plexus had a better prognosis than those who had a closed injury. Polytrauma patients and those with penetrating neck injuries should be assessed to exclude brachial plexus injury. Baseline assessment and early involvement of surgeons with an interest in this area will help select those patients who will benefit from brachial plexus reconstruction. PMID- 11879836 TI - The use of subtalar arthroscopy in open reduction and internal fixation of intra articular calcaneal fractures. AB - Congruency of the subtalar joint is a major predictor of outcome after intra articular calcaneal fractures. Because of the irregular shape of the posterior joint facet, minor steps may be overlooked with direct vision or intra-operative fluoroscopy during open reduction and internal fixation (ORIF). In a preliminary series of 15 subtalar arthroscopies during hardware removal after ORIF of intra articular calcaneal fractures, patients with visible steps of 1 mm or more had significantly inferior functional results compared to those with an intact subtalar joint (117 vs. 158 points with the 200-point score, 71.3 vs. 91.2 points with the Maryland Foot Score, P<0.01). Congruency of the posterior facet was controlled arthroscopically in 47 intra-articular calcaneus fractures after open reduction and temporary K-wire-fixation. In 12 cases (25.5%), despite seemingly correct reduction as judged fluoroscopically, incongruencies in the posterior calcaneal facet were detected and reduction was repeated, temporarily removing the K-wires. After anatomical reduction a standard AO calcaneal plate was fixed to the lateral wall. Intra-operative arthroscopy appears to be useful in detecting minor incongruencies after ORIF of intra-articular calcaneus fractures. In the hands of an experienced surgeon it is less time demanding than intra operative X-rays and more precise than fluoroscopy. PMID- 11879837 TI - The TourniquAid: prevention of chemical burns. PMID- 11879838 TI - Padding under tourniquets in tourniquet controlled surgery: Bruner's ten rules revisited. PMID- 11879839 TI - The unforgettable finger tourniquet. PMID- 11879840 TI - Non-electrical 'burn' from a diathermy pad. PMID- 11879841 TI - Mal-united femoral fractures adjacent to loose total hip arthroplasties. Salvage with impaction grafting. A case report. PMID- 11879842 TI - Hip fracture in adult osteopetriosis. PMID- 11879843 TI - Bilateral symmetrical periarticular trauma of shoulder and hip joints. PMID- 11879844 TI - Topical application of doxepin hydrochloride can reduce the symptoms of complex regional pain syndrome: a case report. PMID- 11879849 TI - Dynamic hip screw: does side make a difference? Effects of clockwise torque on the right and left DHS. PMID- 11879851 TI - Re: Davies et al. Irreducible anterior shoulder dislocation: the greater tuberosity Hill-Sachs lesion. Injury 2000;31:470-1. PMID- 11879852 TI - Hip fractures--the way forward. PMID- 11879853 TI - Time to look beyond MMR in autism research. PMID- 11879854 TI - Heritability of clotting factors and the revival of the prothrombotic state. PMID- 11879855 TI - Maternal deaths among Afghan refugees. PMID- 11879856 TI - The expanding diversity of rotaviruses. PMID- 11879857 TI - Surgeons' knots: old skills, new training. PMID- 11879858 TI - Maternal mortality among Afghan refugees in Pakistan, 1999-2000. AB - BACKGROUND: Estimated at 3.6 million, Afghans are the largest population of refugees in the world. Information on the magnitude, causes, and preventable factors of maternal deaths among Afghan refugees may yield valuable information for prevention. METHODS: Deaths were recorded between Jan 20, 1999, and Aug 31, 2000, during a census carried out in 12 Afghan refugee settlements in Pakistan. Deaths among women of reproductive age (15-49 years) were further investigated by verbal autopsy interviews to determine their cause, risk factors, and preventability, and to ascertain the barriers faced to obtaining health care. FINDINGS: The census identified 134406 Afghan refugees and 1197 deaths; a crude mortality rate of 5.5 (95% CI 5.2-5.8) per thousand population. Among the 66 deaths among women of reproductive age, deaths due to maternal causes (n=27) exceeded any other cause (41% [95% CI 29-53]). 16 liveborn and nine stillborn infants were born to women who died of maternal causes; six of the liveborn infants died after birth. Therefore, 60% (15 of 24) of infants born to these women were either born dead or died after birth. Compared with women who died of non-maternal causes, women who died of maternal causes had a greater number of barriers to health care (p=0.001), and their deaths were more likely to be preventable (p<0.05). INTERPRETATION: Maternal deaths account for a substantial burden of mortality among Afghan refugee women of reproductive age in Pakistan. The high prevalence of barriers to health care access indicates opportunities for reducing maternal deaths in refugee women and their children. PMID- 11879859 TI - An unexpected tattoo. PMID- 11879860 TI - Efficacy of a Cancer Research UK communication skills training model for oncologists: a randomised controlled trial. AB - BACKGROUND: Doctors' communication with patients is commonly hampered by lack of training in this core skill. This study aimed to assess the efficacy of an intensive 3-day training course on communication skills in a randomised controlled trial with a two-by-two factorial design and several outcomes. METHODS: 160 oncologists from 34 UK cancer centres were randomly allocated to four groups: written feedback followed by course; course alone; written feedback alone; and control. At each of two assessment periods, consultations with six to ten consecutive, consenting patients per doctor were videotaped. 2407 patients participated. Outcome measures included objective and subjective ratings made by researchers, doctors, and patients. The primary outcomes were objective improvements after the intervention in key communication skills. Course content included structured feedback, videotape review of consultations, role-play with simulated patients, interactive group demonstrations, and discussion led by a trained facilitator. FINDINGS: In Poisson regression analysis of counts of communication behaviours, course attendance significantly improved key outcomes. The estimated effect sizes corresponded to higher rates of use of focused questions (difference between course attenders [n=80] and non-attenders [n=80] 34%, p=0.003), focused and open questions (27%, p=0.005), expressions of empathy (69%, p=0.003), and appropriate responses to patients' cues (38%, p=0.026), and a 24% lower rate of use of leading questions (p=0.11). There was little evidence for the effectiveness of written feedback. INTERPRETATION: The communication problems of senior doctors working in cancer medicine are not resolved by time and clinical experience. This trial shows that training courses significantly improve key communication skills. More resources should be allocated to address doctors' training needs in this vital area. PMID- 11879861 TI - Corticosteroid injections, physiotherapy, or a wait-and-see policy for lateral epicondylitis: a randomised controlled trial. AB - BACKGROUND: Lateral epicondylitis is generally treated with corticosteroid injections or physiotherapy. Dutch clinical guidelines recommend a wait-and-see policy. We compared the efficacy of these approaches. METHODS: Patients with lateral epicondylitis of at least 6 weeks' duration were recruited by family doctors. We randomly allocated eligible patients to 6 weeks of treatment with corticosteroid injections, physiotherapy, or a wait-and-see policy. Outcome measures included general improvement, severity of the main complaint, pain, elbow disability, and patient satisfaction. Severity of elbow complaints, grip strength, and pressure pain threshold were assessed by a research physiotherapist who was unaware of treatment allocation. We assessed all outcomes at 3, 6, 12, 26, and 52 weeks. The principal analysis was done on an intention-to-treat basis. FINDINGS: We randomly assigned 185 patients. At 6 weeks, corticosteroid injections were significantly better than all other therapy options for all outcome measures. Success rates were 92% (57) compared with 47% (30) for physiotherapy and 32% (19) for wait-and-see policy. However, recurrence rate in the injection group was high. Long-term differences between injections and physiotherapy were significantly in favour of physiotherapy. Success rates at 52 weeks were 69% (43) for injections, 91% (58) for physiotherapy, and 83% (49) for a wait-and-see policy. Physiotherapy had better results than a wait-and-see policy, but differences were not significant. INTERPRETATION: Patients should be properly informed about the advantages and disadvantages of the treatment options for lateral epicondylitis. The decision to treat with physiotherapy or to adopt a wait-and-see policy might depend on available resources, since the relative gain of physiotherapy is small. PMID- 11879862 TI - Angiotensin-converting-enzyme inhibition therapy in altitude polycythaemia: a prospective randomised trial. AB - BACKGROUND: Angiotensin-converting-enzyme (ACE) inhibitors reduce packed cell volume and haemoglobin concentration in polycythaemia that follows renal transplantation, which, like altitude polycythaemia, is an erythropoietin dependent form of polycythaemia. We aimed to establish the effect of ACE inhibitor treatment in people with altitude polycythaemia. METHODS: We did a prospective randomised study in 26 people with altitude polycythaemia (packed cell volume > or = 55%) and 24-h rate of urinary protein excretion greater than 150 mg, who had been referred to the Renal Disease Project in La Paz, Bolivia. 13 participants were assigned 5 mg/day enalapril for 2 years (study group), and 13 no treatment (controls). Blood pressure, packed cell volume and haemoglobin concentration, proteinuria, and renal function were compared by intention-to treat analyses. FINDINGS: Baseline packed cell volume and haemoglobin concentration were positively correlated with bodyweight (p=0.02), systolic (p=0.01) and diastolic (p=0.04) blood pressure, serum creatinine (p=0.009), blood urea (p=0.008), and proteinuria (p=0.003). Systolic and diastolic blood pressure remained stable in the study group, but increased in controls. In study patients, mean (SD) packed cell volume, haemoglobin concentration, and proteinuria fell from 63.5% (4.9) to 56.8% (4.1), p<0.0001; 207 (18) to 164 g/L (13), p<0,0001; and from 358.6 (260.3) to 247.7 mg/24-h (208.2), p<0.002, respectively, but did not change significantly in controls. At 12 and 24 months of follow-up, packed cell volume, haemoglobin concentration, and proteinuria differed significantly between the groups (p<0.0001 for each comparison). In study patients, follow-up changes in packed cell volume (r=0.88, p<0.0001) or haemoglobin concentration (r=0.83, p<0.0001) and proteinuria were strongly correlated. Enalapril was well tolerated by all patients. INTERPRETATION: ACE-inhibition therapy effectively and safely ameliorates altitude polycythaemia and reduces proteinuria. PMID- 11879863 TI - Activation markers of coagulation and fibrinolysis in twins: heritability of the prethrombotic state. AB - BACKGROUND: Activation markers of coagulation and fibrinolysis are increased in individuals at risk of coronary-artery disease and other thrombotic disorders--a condition defined as the prethrombotic state. We aimed to find out the extent to which the prethrombotic state is determined by genetic factors. METHODS: We analysed concentrations of prothrombin, prothrombin fragment 1+2, thrombin antithrombin complex, crosslinked fibrin degradation product D-dimer, and thrombin-activatable fibrinolysis inhibitor by ELISA in 118 monozygotic and 112 dizygotic unselected female twins aged 21-73 years from the St Thomas' UK Adult Twin Registry. We used quantitative genetic-model fitting to estimate heritability. FINDINGS: We found significant heritabilities in concentrations of the activation markers in plasma. Genetic factors contributed 45, 40, and 65% of the variation in concentrations of fragment 1+2, thrombin-antithrombin complex, and D-dimer, respectively. Age was important only in fragment 1+2 concentrations, in which it accounted for 12% of the variation. The remaining variation could be attributed to unique environmental factors. Variation in concentrations of precursor prothrombin in plasma was determined by 57% heritability, and that of zymogen thrombin-activatable fibrinolysis inhibitor showed a very strong genetic component (82%). INTERPRETATION: The activation mechanisms of the coagulation and fibrinolytic systems, and therefore the prethrombotic state, are controlled to a substantial degree by genetic factors. Genes influencing activation of haemostasis are likely to be an important component of the overall thrombotic tendency in the general population. PMID- 11879864 TI - Vitamin D poisoning by table sugar. PMID- 11879865 TI - Surgically obtained sperm, and risk of gestational hypertension and pre eclampsia. AB - The cause of pre-eclampsia is unknown, although a partner-specific immune maladaptation might be involved. We compared rates of pre-eclampsia and gestational hypertension in women whose genital tracts had and had not been exposed to their partner's sperm cells. Our aim was to ascertain whether or not protective partner-specific immune-tolerance is conveyed by sperm cells, rather than seminal fluid. Our findings indicate that, compared with women exposed to their partner's sperm cells and seminal fluid--ie, those treated with in-vitro fertilisation (IVF) or intracytoplasmic sperm injection (ICSI) with ejaculated sperm--the risk of hypertension was doubled and the risk of pre-eclampsia tripled in those never exposed to their partner's sperm--ie, those treated with ICSI done with surgically obtained sperm. PMID- 11879866 TI - Congenital rubella syndrome: a risk in immigrant populations. AB - An infant with congenital rubella syndrome was born to a young mother who had recently arrived in the UK. The infection was not detected before birth, and transmission to another infant was documented. This case highlights the emerging importance of rubella as an imported infection in the developed world and the need to maintain a high index of suspicion for this disorder in recent immigrants from countries with no immunisation programme. Targeted immunisation for such groups is recommended. PMID- 11879867 TI - Hormone replacement therapy and prevention of pressure ulcers and venous leg ulcers. AB - Pressure ulcers and venous leg ulcers are common chronic wounds. Oestrogens in the form of hormone replacement therapy (HRT) might have an effect on wound healing, but this possibility has not been studied in detail. Using a case-cohort study including elderly patients in the UK General Practice Research Database, we showed that patients who received HRT were less likely to develop a venous leg ulcer (age-adjusted relative risk 0.65 [95% CI 0.61-0.69]) or a pressure ulcer (0.68 [0.62-0.76]) than those who did not use HRT. Therefore, we believe that HRT could be beneficial for the prevention of these wounds. PMID- 11879868 TI - Role of Purkinje conducting system in triggering of idiopathic ventricular fibrillation. AB - Ventricular fibrillation is the main mechanism of sudden cardiac death, but the source of its spontaneous initiation has not been mapped. 16 patients were investigated by electrography and radiofrequency ablation after resuscitation from recurrent idiopathic ventricular fibrillation. Triggers of ventricular fibrillation originated from various locations within the Purkinje system in 12 patients and from the ordinary myocardial muscle in four. The accuracy of mapping was confirmed by acute elimination of triggers by radiofrequency delivery, and there was no recurrence of ventricular fibrillation in 14 patients. Long-term follow-up is necessary to establish that ablation is curative and avoids use of a defibrillator. PMID- 11879869 TI - Fears that security rules will impede US and UK science. PMID- 11879872 TI - Better surveillance needed for second colorectal cancers. PMID- 11879873 TI - Neisseria gonorrhoeae evades host immunity by switching off T lymphocytes. PMID- 11879874 TI - Thin mice and fat mice respond differently to appetite suppressant. PMID- 11879875 TI - Would you want someone interfering with your RNA? PMID- 11879876 TI - "Inside" Australia's Woomera detention centre. PMID- 11879877 TI - Canada outlines 10-year research strategy that will "take on the world". PMID- 11879879 TI - Ireland addresses EU call for short working week for doctors. PMID- 11879880 TI - Pakistan's province limits private practice in public hospitals. PMID- 11879881 TI - Pan African group takes lead against the tsetse fly. PMID- 11879882 TI - The muscular dystrophies. AB - The muscular dystrophies are inherited myogenic disorders characterised by progressive muscle wasting and weakness of variable distribution and severity. They can be subdivided into several groups, including congenital forms, in accordance with the distribution of predominant muscle weakness: Duchenne and Becker; Emery-Dreifuss; distal; facioscapulohumeral; oculopharyngeal; and limb girdle which is the most heterogeneous group. In several dystrophies the heart can be seriously affected, sometimes in the absence of clinically significant weakness. The genes and their protein products that cause most of these disorders have now been identified. This information is essential to establish an accurate diagnosis and for reliable genetic counselling and prenatal diagnosis. There is, as yet, no way of greatly affecting the long-term course of any of these diseases. However, advances in gene manipulation and stem-cell therapy suggest cautious optimism for finding an effective treatment in the not-too-distant future. PMID- 11879883 TI - Uses of error. PMID- 11879884 TI - Blinding in randomised trials: hiding who got what. AB - Blinding embodies a rich history spanning over two centuries. Most researchers worldwide understand blinding terminology, but confusion lurks beyond a general comprehension. Terms such as single blind, double blind, and triple blind mean different things to different people. Moreover, many medical researchers confuse blinding with allocation concealment. Such confusion indicates misunderstandings of both. The term blinding refers to keeping trial participants, investigators (usually health-care providers), or assessors (those collecting outcome data) unaware of the assigned intervention, so that they will not be influenced by that knowledge. Blinding usually reduces differential assessment of outcomes (information bias), but can also improve compliance and retention of trial participants while reducing biased supplemental care or treatment (sometimes called co-intervention). Many investigators and readers naively consider a randomised trial as high quality simply because it is double blind, as if double blinding is the sine qua non of a randomised controlled trial. Although double blinding (blinding investigators, participants, and outcome assessors) indicates a strong design, trials that are not double blinded should not automatically be deemed inferior. Rather than solely relying on terminology like double blinding, researchers should explicitly state who was blinded, and how. We recommend placing greater credence in results when investigators at least blind outcome assessments, except with objective outcomes, such as death, which leave little room for bias. If investigators properly report their blinding efforts, readers can judge them. Unfortunately, many articles do not contain proper reporting. If an article claims blinding without any accompanying clarification, readers should remain sceptical about its effect on bias reduction. PMID- 11879885 TI - Now or never: the case for measuring maternal mortality. PMID- 11879886 TI - Autism, bowel inflammation, and measles. PMID- 11879887 TI - Mammographic screening: no reliable supporting evidence? PMID- 11879889 TI - Bovine tuberculosis: milk and meat safety. PMID- 11879890 TI - Use of misoprostol in third stage of labour. PMID- 11879892 TI - Use of misoprostol in third stage of labour. PMID- 11879891 TI - Use of misoprostol in third stage of labour. PMID- 11879893 TI - Use of misoprostol in third stage of labour. PMID- 11879895 TI - Anthrax inhalation and lethal human infection. PMID- 11879896 TI - Anthrax inhalation and lethal human infection. PMID- 11879897 TI - The virgin cleansing myth: cases of child rape are not exotic. PMID- 11879898 TI - Statins and myopathy. PMID- 11879899 TI - Re-emergence of ebola haemorrhagic fever in Gabon. PMID- 11879900 TI - Transplantation in chronic myeloid leukaemia. PMID- 11879901 TI - Global issues in medical education. PMID- 11879902 TI - Strengthening research capacity. PMID- 11879903 TI - On prion disease and form. PMID- 11879904 TI - Global issues in medical education. PMID- 11879905 TI - Newspaper consultations. PMID- 11879908 TI - Tools of the trade. The Smith-Clarke respirator. PMID- 11879913 TI - Should we always get the words right? PMID- 11879915 TI - Stilbene photochrome-fluorescence-spin molecules: covalent immobilization on silica plate and applications as redox and viscosity probes. AB - We report herein on the development of a new photochrome-fluorescence-spin method for the quantitative analysis of the redox status and viscosity of a medium. The method of the viscosity measurement is based on the use of double fluorescence nitroxide molecules. In such hybrid compounds the nitroxide moiety quenches the fluorescence of the fluorophore (stilbene moiety). The reduction of nitroxide by an antioxidant (ascorbic acid) causes a rise of fluorescence of the fluorophore. The rate constant of the stilbene fragment photoisomerization in such systems is dependent upon the viscosity of the media. The synthesized dual stilbene nitroxide probe was covalently immobilized onto the surface of a quartz plate as an eventual fiber-optic sensor. The immobilization procedure included a cyanogen bromide surface activation followed by smoothing with a protein tether. The rate of fluorescence change was monitored in aqueous-glycerol solutions of different viscosities and content of ascorbic acid. Good correlation was found: (a) between the concentration of ascorbic acid in the sample and the rate of fluorescence increase due to the reduction of the nitroxide moiety, and (b) between the rate constant of photoisomerization and the viscosity of the media. Appropriate calibration would make the determination of the viscosity of a media possible (in a range 1-500 cP), as well as ascorbate content, in a range (1-9) x 10(-4) M, with fast single measurement. PMID- 11879916 TI - Direct on air sampling filter quantification of cat allergen. AB - A direct on sampling filter in solution (DOSIS) method for quantification of airborne cat allergens has been developed. In this method, the allergens firmly adsorbed to a porous polytetrafluoroethylene filter are reacted with specific antibodies conjugated to alkaline phosphatase, generating a matrix-bound allergen antibody-phosphatase complex. The treated filter is subsequently floated on a commercially available chemiluminescent phosphatase substrate solution. Aliquots of this solution are removed and analyzed luminometrically. The light intensity of the product is linearly related to the amount of allergen over a large mass range, 0-100 SQ units (1 SQ unit is about 146 pg of the allergenic protein Fel d 1). DOSIS demonstrated intra- and interassay precisions of 9% and 8% and 14% and 21% for the levels 4 and 20 SQ units per filter, respectively. The limit of quantification was estimated to 0.4 SQ units (58 pg Fel d 1) of cat allergen per filter. Application of DOSIS to analysis of cat allergen concentrations of indoor air in homes with and without cats revealed, on average, a six times higher concentration in the former (142 SQ units/m(3)) as compared to the latter (24 SQ units/m(3)). The recorded concentrations for airborne cat allergen in homes with cats are in accordance with previously reported figures. Allergen-specifically stained sampling filters revealed the particulate nature of airborne cat allergen which seemed predominantly to be carried by numerous large dust particles. PMID- 11879917 TI - NMR study of cyclic peptides with renin inhibitor activity. AB - Several cyclic analogues of renin inhibitors, based on Glu-D-Phe-Lys motif have been investigated by NMR spectroscopy and molecular dynamics calculations (MD). The 15 membered macrocycle, resulting from Glu and Lys side-chain cyclization, exhibits conformational preference. The structural evidence from NMR shows the presence of hydrogen bond between Lys NH and Glu side-chain carbonyl, resulting in a 10 membered pseudo beta-turn-like structure. The structure of the cyclic moiety is similar in all the peptides, which takes at least two conformations around Calpha-Cbeta in Glu side chain. The restrained MD calculations further support such observations and show that the macrocycle is fairly rigid, with two conformations about the Glu Calpha-Cbeta bond. The linear peptide appendages, which are essential for activity in cyclic peptides, show an extended structure in the beta-region of Ramchandran plot. These calculations also demonstrate that for the most active peptide, two major conformers each exist about the Calpha-CO bond of the Lys, D-Trp and Leu residues. In this peptide, the cyclic moiety presents a negatively charged surface formed due to the carbonyl oxygens, which are thus available to form hydrogen bonds with the receptor. The linear fragment presents further binding sites with a surface which has the hydrophobic side chains of D-Trp, Leu and D-Met on one side and carbonyls on the other side. PMID- 11879918 TI - Sensitive fluorescent detection of protein on nylon membranes. AB - Detection of antigen immobilized on membranes, as in Western transfers and dot enzyme linked immunosorbent assays (ELISAs), often employ antibody-enzyme conjugates and chemiluminescent or precipitated colored reaction products. Although chemiluminescent markers are sensitive, they are time-consuming because of their required exposure to X-ray film and the presence of background artifacts sometimes limits their use. This report demonstrates that direct fluorescent detection technique using nylon membranes that has higher sensitivity than chemiluminescent methods is easier to perform and has a uniform, low background. An alkaline phosphatase conjugated antibody was compared with antibody conjugated to a fluorescent phycobiliprotein (allophycocyanin) for sensitivity in both Western transfers and dot ELISA assays using mouse IgG as the membrane-bound antigen. Direct fluorescent detection of antigen-antibody complexes on positively charged nylon membrane provided better sensitivity and lower background than similar conditions using enzyme amplification and chemiluminescent detection on either nylon or PVDF membranes. Processing time was reduced by the elimination of steps associated with substrate incubation, washing and X-ray film exposures required for chemiluminescence detection. These data support the view that direct fluorescent detection can represent a significant improvement in assay sensitivity and reduction in time compared with more traditional chemiluminescent detection techniques employed in the conduct of Western transfers and dot ELISA studies. PMID- 11879919 TI - Direct detection of isotopically labeled metabolites bound to a protein microarray using a charge-coupled device. AB - A charge-coupled device (CCD) was used to quantitatively detect isotope-labeled ligands bound to a protein microarray. Protein microarrays with protein dots, 10 50 microm in diameter, were fabricated on an aluminized Mylar film using an electrospray deposition technique. Proteins in dots were immobilized by cross linking in glutaraldehyde vapor. After contact with solutions of isotope-labeled metabolites, the protein microarrays were washed, dried and placed face down onto the surface of a standard B/W video CCD chip with the protective window removed. We show here that such a simple inexpensive CCD detector can be used to quantify distribution of 14C and other radioactive isotopes on microarrays. PMID- 11879920 TI - An extended boiling method for small-scale preparation of plasmid DNA tailored to long-range automated sequencing. AB - Automated fluorescence sequencing depends on high-quality plasmid DNA, which is conveniently prepared by minipreparation procedures. While those procedures are effective for high-copy number plasmids, purity and yields of low-copy number plasmids are often not sufficient to achieve reasonable sequencing results. Here, we describe a reproducible and cheap procedure for the small-scale preparation of plasmid DNA, which is based on the original Holmes and Quigley protocol, comprising a boiling and two selective precipitation steps. Besides various other modifications, this procedure utilizes polyethylene glycol (PEG) precipitation as a key step to further purify plasmid DNA tailored to automated fluorescence sequencing. Independent of the plasmid size and copy number, the modified procedure yields plasmid DNA, which gives average reading lengths of 800 and more bases with a standard fluorescence cycle sequencing protocol. To demonstrate the efficiency and reproducibility of the method, sequencing data of various human interleukin-6 gene variants cloned in different vectors are presented. This procedure offers an economical alternative to commercial miniprep kits, utilizing silica resins or anion-exchanger matrices and, moreover, is more reliable and consistent with respect to reading lengths and accuracy in automated fluorescence sequencing. PMID- 11879921 TI - Optimisation and validation of an angiotensin-converting enzyme inhibition assay for the screening of bioactive peptides. AB - Angiotensin-converting enzyme (ACE) plays a major role in the regulation of blood pressure. A diagnostic assay to measure angiotensin-converting enzyme (ACE) activity was transformed into an enzyme inhibition assay and optimised, which led to a more sensitive and less expensive assay. By this spectrophotometric method, ACE inhibition is measured using the substrate furanacryloyl-Phe-Gly-Gly and as ACE source rabbit lung acetone extract. The optimised as well as the original ACE inhibition assay were used to verify the ACE inhibitory activity of captopril. The ACE inhibition assay was further validated by enalapril, its active derivative enalaprilat and the ACE-inhibitory peptide Ala-Leu-Pro-Met-His-Ile Arg, corresponding to a tryptic fragment of bovine beta-lactoglobulin. Sigmoid curves could be fit adequately to the data points representing ACE inhibition in function of inhibitor concentration. IC(50) values for these compounds corresponded well with literature data. Furthermore, pea and whey protein hydrolysates obtained by digestion with trypsin showed ACE inhibitory activity in the ACE inhibition assay. Hence, this optimised assay is suitable to screen for ACE inhibitory peptides derived from food proteins with a possible antihypertensive effect in vivo. PMID- 11879922 TI - Mutation analysis of MLH1 and MSH2 genes performed by denaturing high-performance liquid chromatography. AB - Mutation analysis of large genes, such as MSH2 and MLH1, is time-consuming and expensive. We investigated the sensitivity and specificity of DHPLC analysis for the detection of mutations within both MSH2 and MLH1. Studies included a series of 46 patients affected by colorectal cancer from HNPCC families. We confirmed 19 changes previously identified by DNA sequencing and, in a blind study, an additional 16 rare alterations including four mutations not previously described. Generally, false negative results were not observed. Elution profiles were highly characteristic for a given change and in 98.5% cases allowed the distinction between novel alterations and previously identified mutations and polymorphisms. For the detection of changes in almost all amplicons, it was sufficient to use just one denaturing temperature. DHPLC was confirmed to be highly sensitive, specific and a cost-effective technique with particularly high potential for the detection of MSH2 and MLH1 gene mutations in the diagnostic setting. PMID- 11879923 TI - Antimicrobial guidelines for the treatment of acute bacterial rhinosinusitis in immunocompetent children. AB - Acute rhinosinusitis represents a condition for which educational efforts could help minimize the inappropriate use of antibiotics, particularly for children. The majority of acute rhinosinusitis cases are of viral etiology and thus, are self limiting. Although bacterial infection complicates a small number of cases, the lack of accessibility to the sinus, the limitations of diagnostic modalities and the lack of specificity among signs and symptoms often make it difficult to determine when bacterial infection occurs. Furthermore, antimicrobial resistance among the pathogens that frequently cause bacterial infection complicates the election of empiric therapy. The Sinus and Allergy Health Partnership recently developed and published antimicrobial guidelines to provide practitioners in the US with recommendations for the diagnosis and treatment of acute bacterial rhinosinusitis. The purpose of this paper is to review the rationale behind the development of these guidelines and how they apply to the management of acute bacterial rhinosinusitis in children. PMID- 11879924 TI - Tonsillectomy for biopsy in children with unilateral tonsillar enlargement. AB - OBJECTIVE: Unilateral tonsillar enlargement (UTE) may be a sign of underlying malignancy and tonsillectomy is often recommended for histology. The limited evidence available suggests that the incidence of malignancy in children with UTE is very low, and that in many cases, the apparent enlargement is due to asymmetry of the mucosa of the tonsillar pillars. The purpose of this study was to evaluate the necessity of tonsillectomy as a routine practice for every case of UTE. METHODS: We attempted to identify every child (age < 16 years) undergoing tonsillectomy in our unit from January 1991 to January 2000 where the indication for surgery was UTE. Patients were excluded if the primary indication for surgery was anything other than UTE. The case notes and pathology records were reviewed. RESULTS: Exactly 47 children were identified, aged between 4 and 15 years of age (mean 9.5 years), of whom 29 (62%) were girls. In all cases, the tonsillar asymmetry was noted by the otolaryngologist, but had not been noted by the patient or parents, or commented on by the referring General Practitioner. Eighteen of the children (38%) had no history of sore throats. No malignancies were encountered. When the tonsils were measured by the pathologist after excision, the actual degree of asymmetry in size ranged from 0 to 25 mm. In seven cases (15%), the clinically larger tonsil was actually the smaller of the two when examined by the pathologist. In 17 cases (36%) there was no difference in size at all. CONCLUSIONS: We conclude that apparent UTE is often spurious, and that many unnecessary tonsillectomies are performed when UTE is taken as an indication for surgery in the absence of any other suspicious features. Tonsillectomy carries risks and clinical observation may be prudent where clinical suspicion is low. PMID- 11879925 TI - Colonization rate of bacteria in the throat of healthy infants. AB - OBJECTIVE: the human throat is a major ecological site for various bacteria that can reach neighbouring sterile sites and cause mild infections or invasive diseases. The aim of this study was to investigate the carriage rate of several potential pathogens in the throat of healthy children under the age of 2 years. METHODS: cultures were taken from the tonsils of 1000 healthy infants aged 1-24 months attending well-baby clinics, who had not received antibiotic therapy during the preceding 14 days. RESULTS: one hundred and ninety-eight (19.8%) cultures were positive. Thirteen (1.3%) cultures were positive for beta haemolytic Streptococcus group A, 23 (2.3%) for Streptococcus pneumoniae. In 28 (2.8%) and 24 (2.4%) cultures, respectively, Haemophilus influenzae Type b and non-typeable Haemophilus influenzae were recovered. The commonest bacterium found was Staphylococcus aureus (99 positive cultures). Eleven children carried two species of bacteria and from one 6-month-old child three species were isolated concurrently. CONCLUSIONS: it is concluded that children younger than 2 years of age can be carriers of several types of pathogenic bacteria. In contrast to many other studies, in this study beta-haemolytic Streptococcus group A was isolated from the tonsils of children younger than 1 year of age. PMID- 11879926 TI - A study on the prevalence of accessory auricle anomaly in Turkey. AB - OBJECTIVE: Accessory auricular anomaly is a small elevation of skin containing a bar of elastic cartilage localized most commonly just anterior to the tragus or ascending crus of the helix. The anomaly may exist isolated or may be associated with other congenital anomalies of the first arch. The purpose in this study is to detect prevalence of accessory auricle in Turkey and find out whether it is associated with other craniofacial anomalies or hearing loss. METHODS: The study was performed on 850 children from the age of 7 to 9 during a screening program in primary schools. Complete otolaryngologic examination and acoustic reflectometry measurements were performed on all the children. Full physical examination, tympanometric and audiometric evaluation and EEG measurements were added to the cases with accessory auricle anomaly. RESULTS: Among 850 children examined, four had accessory auricle anomaly and prevalence of the anomaly was calculated as 0.47% (95% confidence interval (CI), 0.13-1.20%). Children were developmentally normal, and no other congenital craniofacial or systemic anomaly was detected in any of the cases. Further, tympanometric, audiometric evaluations and EEG tests were in normal limits. CONCLUSIONS: In this study, the prevalence rate of the condition was calculated as 0.47%. Although one study from China reported this prevalence as 0.22%, the difference between the reported prevalances was not statistically significant. Further, although external ear anomalies may present together with cranifacial anomalies and neurologic disorders like epilepsy, neuromotor retardation and EEG disorders, in our cases, mental and motor development was normal and epilepsy history or abnormal EEG patterns do not exist. On the other hand, no hearing loss was found to be associated with accessory auricles. PMID- 11879927 TI - The shortened cochlea: its classification and histopathologic features. AB - INTRODUCTION: The term 'Mondini dysplasia' has been used to describe virtually any congenital abnormality of the osseous labyrinth resulting in confusion and seemingly contradictory observations and conclusions about this type of deformity. The purpose of this study is to histopathologically classify and describe temporal bones whose cochleas have less than 2.5 turns. METHODS: Of the 1800 temporal bones in our collection, 21 from 12 cases were found to have cochleas with less than 2.5 cochlear turns. Ages ranged from stillborn to 50 years. Temporal bones were harvested at autopsy, processed and embedded in celloidin. Sections were cut at a thickness of 20 microm and every 10th section stained with hematoxylin-eosin and examined using light microscopy. The number of turns, length of cochlea, integrity of cochlear base, length of modiolus, abnormalities of the semicircular canals and vestibule, enlargement of the vestibular aqueduct and middle ears were documented. Twenty-one temporal bones from age-matched patients without cochlear deformities were used as controls for modiolar length measurements. RESULTS: Malformation of the shortened cochlea was histopathologically classified into three groups as follows: (1) Common cavity, cochlear dysplasia (one ear)--severe dysplasia of the cochlea without a complete basal turn; (2) Mondini dysplasia (11 ears)--1.5 cochlear turns, a complete basal turn, an incomplete or absent interscalar septum and a complete bone at the base of the modiolus; and (3) Mondini-like dysplasia type A (five ears)--2 turns to the cochlea including a complete basal turn and complete bone at the base of the modiolus; and type B (four ears)--1.5-2 turns to the cochlea, hypoplasia of or a missing bone at the base of the modiolus (either with or without a communication between the internal auditory canal and the cochlea) and a complete basal turn. CONCLUSION: The range of congenital malformations in short cochlea is highly variable. Fundamental to the accurate evaluation of a labyrinthine anomaly, malformations of the inner ear should be classified according to the findings in the labyrinth. We suggest the use of common cavity cochlear dysplasia, Mondini dysplasia and Mondini-like dysplasia to describe these variable anomalies. PMID- 11879928 TI - Nasal septum deformities in children and adolescents: a cross sectional study of children from Zagreb, Croatia. AB - OBJECTIVE: Numerous epidemiologic studies on the prevalence of nasal septum deformities in children have been performed over the last three decades. As these studies were performed in children of various age groups and used different classifications of septal deformities (without detailed morphologic systematization), it is no surprise that the results differ greatly from study to study. The purpose of the study was to evaluate clinical data on the total prevalence of nasal septum deformities and particular types of deformity in children and adolescents. METHODS: The study population consisted of 1797 randomly selected subjects divided into pre-school (aged 2-6), primary school (aged 7-14), secondary school (aged 15-18) and university (aged 19-22) groups. The native state was determined by means of anterior rhinoscopy without previous application of vasoconstrictive drugs. The observed pathologic septal deformities were classified into seven types according to Mladina's classification. A straight septum was designated as S. The prevalence of septum deformities in the population was calculated with 95% probability. Differences with respect to sex and types of deformity were tested by chi(2)-test. RESULTS: The prevalence of nasal septum deformities according to age groups was 28.0% in the 2-6 group, 21.1% in the 7-14 group, 40.6% in the 15--18 group and 41.8% in the 19-22 group. The distribution of the seven types of septal deformity was 51.1, 23.5, 5, 0.8, 10.0, 9.0 and 0.6%, respectively. The mean values (%) and 95% confidence intervals for the seven types of septal deformity were 14.7 (13.1-16.4%), 6.8 (5.6-7.9%), 1.4 (0.9-1.9%), 0.2 (0.0-0.4%), 2.9 (2.1-3.7%), 2.6 (1.9-3.4%) and 0.2 (0.0-0.4%), respectively. Total distribution in gender showed no difference (P = 0.102). CONCLUSIONS: In the youngest age group (2-6 years), types 1 and 2 (deformities of anterior septal segments) were exclusively found, whereas types 5 and 6 were found in older age groups (become visible during and after the puberty). Types 1 and 2 are characteristic septal deformities for small children. Since, septal deformities can affect the growth and development of the maxilla and vice versa, the authors recommend examination of the nasal septum by an rhinologist who will be a part of a team performing the regular systematic health examination of children. PMID- 11879929 TI - Treatment of chronic suppurative otitis media with ofloxacin in hydroxypropyl methylcellulose ear drops: a clinical/bacteriological study in a rural area of Malawi. AB - Chronic suppurative otitis media in young children is a major problem in Africa, with socio-economic consequences at a later age. Common treatment regimens with antibiotics are expensive and often not practically feasible. Therefore, a project was started to develop a low-cost and effective treatment in a rural area of Malawi by studying the clinical efficacy of an inexpensive application regimen of ofloxacin (0.075%) in hydroxypropyl methylcellulose (1.5%) ear drops. In earlier studies with this treatment regimen, it was possible to cure approximately 70% of ears. The aim of this study was to find out whether the bacteriological spectrum cultured from wet ears before and after treatment, and patterns of resistance to antibiotics, played a role in the percentage of cures. Patients with long-standing chronic suppurative otitis media were clinically assessed and treated with suction cleaning and instillation of ear drops on days 1, 3, 7 and 10. Bacterial swabs were taken for culture and sensitivity tests for ofloxacin were on days 1 and 10 from the ears that were still discharging. After 21 weeks, the ears were assessed again clinically. Clinical cure was considered to be complete cessation of otorrhea. Ninety of 104 tested patients (124 ears) completed the study. About 73% of the ears had become dry by day 10. This dropped to 42% after 21 weeks. Before treatment, most ears (91%) harbored fecal bacteria, Proteus mirabilis (74%) and enterococci (60%) being the most frequently isolated microbes. The second group of frequently cultured bacteria were water bacteria e.g. Pseudomonas species and other non-fermenters (69%), whereas the classical otitis media pathogens were detected only in 15% of ears. Before treatment, 9.7% of strains were resistant to ofloxacin, most (30/35) of which were cultured from ears that were eventually cured. After treatment, fecal and water bacteria were still the most frequently found, with 36% new strains and an overall sensitivity to ofloxacin of 58%. Bacterial resistance did not appear to play an important role in the outcome of treatment. These data rather suggest a very high risk of infection due to poor hygiene conditions. Medical treatment can only have a longer-lasting effect if accompanied by community-based programs that focus on improvement of hygiene. A public health approach is necessary alongside a medical approach for the management of CSOM. PMID- 11879930 TI - Rare case of spontaneous closure of tracheo-esophageal fistula secondary to disc battery ingestion. AB - Accidental ingestion of foreign bodies by children is not an uncommon event. Now a day, with increasing accessibility of electronic toys and devices to the children, ingestion of miniature size batteries is on the rise [J. Am. Med. Assoc., 249, (1983), 2495]. Unlike the usual foreign bodies, ingestion of button alkaline batteries may be associated with serious complications if not removed early [Am. J. Otolaryngol., 21 (2000) 333]. We report a rare and interesting case, in which a child developed tracheo-esopbageal fistula secondary to disc battery ingestion, and importantly, its conservative management resulting in its spontaneous closure. PMID- 11879931 TI - Monophasic synovial sarcoma of the neck in an 8-year-old girl resembling a thyroglossal duct cyst. AB - Synovial sarcomas of the head and neck are rare. Typically, they are localized laterally in the parapharyngeal space. We report the case of an 8-year-old girl with a monophasic round cell synovial sarcoma of the anterior neck, clinically resembling a thyroglossal duct cyst. Histologic, immunohistochemic and cytogenetic findings are presented with a brief review of the literature. This case reaffirms the importance of considering malignant neoplasms in the differential diagnosis of pediatric neck masses. PMID- 11879932 TI - Parotid gland mucoepidermoid carcinoma in a 4-year-old child. AB - Salivary gland tumors account for less than 5% of the head and neck neoplasms. Among them, mucoepidermoid carcinoma is the most common malignant salivary gland tumor. About 45% of mucoepidermoid carcinomas occur in the parotid gland, and appear around the fifth decade of life, being unusual in children under 10 years. We report a case of a parotid mass arising in a 4-year-old female, who had no lateral adenopathy. After histological examination, the diagnosis was of a low grade mucoepidermoid carcinoma of the parotid gland. Three years after surgery no recurrence was observed. As a conclusion, although rare, the presence of a parotid mass with progressive growth in children could correspond to a neoplasm. PMID- 11879933 TI - Actinomycosis of tonsil masquerading as tumour in a 12-year old child. AB - A case of unusual presentation of actinomycosis of tonsil causing massive unilateral enlargement, mimicking a tumor in a 12-year old girl is reported. PMID- 11879934 TI - The birds of Kesterson Reservoir: a historical perspective. AB - Beginning in the late 1970s, Kesterson Reservoir was used for disposal of subsurface drainage from agricultural fields in California's San Joaquin Valley. During 1983-1985, studies were conducted to evaluate the effects of chemicals in this agricultural drainwater on aquatic birds using Kesterson Reservoir. These studies included analyses of food-chain biota (such as plants, aquatic invertebrates, and fish) and bird tissues or eggs, as well as measuring adverse effects on health and reproduction of the birds. Results of the integrated set of field and experimental studies showed that selenium was the only chemical found at concentrations high enough to cause the adverse effects on bird health or reproduction that were observed. This article provides a summary of the field studies conducted at Kesterson Reservoir (and some of the related field and experimental studies conducted elsewhere) to evaluate the effects of irrigation drainage water contaminants on aquatic birds. PMID- 11879935 TI - Role of selenium toxicity and oxidative stress in aquatic birds. AB - Adverse effects of selenium (Se) in wild aquatic birds have been documented as a consequence of pollution of the aquatic environment by subsurface agricultural drainwater and other sources. These effects include mortality, impaired reproduction with teratogenesis, reduced growth, histopathological lesions and alterations in hepatic glutathione metabolism. A review is provided, relating adverse biological effects of Se in aquatic birds to altered glutathione metabolism and oxidative stress. Laboratory studies, mainly with an organic form of Se, selenomethionine, have revealed oxidative stress in different stages of the mallard (Anas platyrhynchos) life cycle. As dietary and tissue concentrations of Se increase, increases in plasma and hepatic GSH peroxidase activities occur, followed by dose-dependent increases in the ratio of hepatic oxidized to reduced glutathione (GSSG:GSH) and ultimately hepatic lipid peroxidation measured as an increase in thiobarbituric acid reactive substances (TBARS). One or more of these oxidative effects were associated with teratogenesis (4.6 ppm wet weight Se in eggs), reduced growth in ducklings (15 ppm Se in liver), diminished immune function (5 ppm Se in liver) and histopathological lesions (29 ppm Se in liver) in adults. Manifestations of Se-related effects on glutathione metabolism were also apparent in field studies in seven species of aquatic birds. Reduced growth and possibly immune function but increased liver:body weight and hepatic GSSG:GSH ratios were apparent in american avocet (Recurvirostra americana) hatchlings from eggs containing 9 ppm Se. In black-necked stilts (Himantopus mexicanus), which contained somewhat lower Se concentrations, a decrease in hepatic GSH was apparent with few other effects. In adult American coots (Fulica americana), signs of Se toxicosis included emaciation, abnormal feather loss and histopathological lesions. Mean liver concentrations of 28 ppm Se (ww) in the coots were associated with elevated hepatic GSH peroxidase, depletion of hepatic protein bound thiols and total thiols, but a small increase in GSH. Diving ducks in the San Francisco Bay area exhibited a positive correlation between hepatic Se concentration and GSH peroxidase activity (r=0.63, P<0.05), but a negative correlation between hepatic Se and GSH concentration (r=-0.740, P<0.05). In willets (Catoptrophorus semipalmatus) from the San Diego area, positive correlations occurred between hepatic Se concentration and GSSG (r=0.70, P<0.001), GSSG:GSH ratio, and TBARS. In emperor geese (Chen canagica) from western Alaska, blood levels of up to 9.4 ppm occurred and were associated with increased plasma GSH peroxidase activity (r=0.62, P<0.001), but with decreased plasma GSSG reductase activity. When evaluating Se toxicity, interactive nutritional factors, including other elements and dietary protein, should also be taken into consideration. Further studies are needed to examine the relationship between different forms of environmentally occurring selenium, arsenic and mercury on reproduction, hepatotoxicity and immune function of aquatic birds. Further selenium nutritional interaction studies may also help to illucidate the mechanism of selenium induced teratogenesis, by optimizing GSH and other antioxidant defense mechanisms in a manner that would stabilize or raise the cell's threshold for susceptibility to toxic attack from excess selenium. It is concluded that Se-related manifestations of oxidative stress may serve as useful bioindicators of Se exposure and toxicity in wild aquatic birds. PMID- 11879936 TI - Selenium toxicity: cause and effects in aquatic birds. AB - There are several manners in which selenium may express its toxicity: (1) an important mechanism appears to involve the formation of CH(3)Se(minus sign) which either enters a redox cycle and generates superoxide and oxidative stress, or forms free radicals that bind to and inhibit important enzymes and proteins. (2) Excess selenium as selenocysteine results in inhibition of selenium methylation metabolism. As a consequence, concentrations of hydrogen selenide, an intermediate metabolite, accumulate in animals and are hepatotoxic, possibly causing other selenium-related adverse effects. (3) It is also possible that the presence of excess selenium analogs of sulfur-containing enzymes and structural proteins play a role in avian teratogenesis. L-selenomethionine is the most likely major dietary form of selenium encountered by aquatic birds, with lesser amounts of L-selenocysteine ingested from aquatic animal foods. The literature is suggestive that L-selenomethionine is not any more toxic to adult birds than other animals. L-Selenomethionine accumulates in tissue protein of adult birds and in the protein of egg white as would be expected to occur in animals. There is no suggestion from the literature that the levels of L-selenomethionine that would be expected to accumulate in eggs in the absence of environmental concentration of selenium pose harm to the developing embryo. For several species of aquatic birds, levels of Se as selenomethionine in the egg above 3 ppm on a wet weight basis result in reduced hatchability and deformed embryos. The toxicity of L-selenomethionine injected directly into eggs is greater than that found from the entry of L-selenomethionine into the egg from the normal adult diet. This suggests that there is unusual if not abnormal metabolism of L selenomethionine in the embryo not seen when L-selenomethionine is present in egg white protein where it likely serves as a source of selenium for glutathione peroxidase synthesis in the developing aquatic chick. PMID- 11879937 TI - Symptoms and implications of selenium toxicity in fish: the Belews Lake case example. AB - Belews Lake, North Carolina was contaminated by selenium in wastewater from a coal-fired power plant during the mid-1970s, and toxic impacts to the resident fish community (20 species) were studied for over two decades. Symptoms of chronic selenium poisoning in Belews Lake fish included, (1) telangiectasia (swelling) of gill lamellae; (2) elevated lymphocytes; (3) reduced hematocrit and hemoglobin (anemia); (4) corneal cataracts; (5) exopthalmus (popeye); (6) pathological alterations in liver, kidney, heart, and ovary (e.g. vacuolization of parenchymal hepatocytes, intracapillary proliferative glomerulonephritis, severe pericarditis and myocarditis, necrotic and ruptured mature egg follicles); (7) reproductive failure (reduced production of viable eggs due to ovarian pathology, and post-hatch mortality due to bioaccumulation of selenium in eggs); and (8) teratogenic deformities of the spine, head, mouth, and fins. Important principles of selenium cycling and toxicity were documented in the Belews Lake studies. Selenium poisoning in fish can be 'invisible', because, the primary point of impact is the egg, which receives selenium from the female's diet (whether consumed in organic or inorganic forms), and stores it until hatching, whereupon it is metabolized by the developing fish. If concentrations in eggs are great enough (about 10 microg/g or greater) biochemical functions may be disrupted, and teratogenic deformity and death may occur. Adult fish can survive and appear healthy despite the fact that extensive reproductive failure is occurring--19 of the 20 species in Belews Lake were eliminated as a result of this insidious mode of toxicity. Bioaccumulation in aquatic food chains causes otherwise harmless concentrations of selenium to reach toxic levels, and the selenium in contaminated sediments can be cycled into food chains for decades. The lessons learned from Belews Lake provide information useful for protecting aquatic ecosystems as new selenium issues emerge. PMID- 11879938 TI - Increased selenium threat as a result of invasion of the exotic bivalve Potamocorbula amurensis into the San Francisco Bay-Delta. AB - Following the aggressive invasion of the bivalve, Potamocorbula amurensis, in the San Francisco Bay-Delta in 1986, selenium contamination in the benthic food web increased. Concentrations in this dominant (exotic) bivalve in North Bay were three times higher in 1995-1997 than in earlier studies, and 1990 concentrations in benthic predators (sturgeon and diving ducks) were also higher than in 1986. The contamination was widespread, varied seasonally and was greater in P. amurensis than in co-occurring and transplanted species. Selenium concentrations in the water column of the Bay were enriched relative to the Sacramento River but were not as high as observed in many contaminated aquatic environments. Total Se concentrations in the dissolved phase never exceeded 0.3 microg Se per l in 1995 and 1996; Se concentrations on particulate material ranged from 0.5 to 2.0 microg Se per g dry weight (dw) in the Bay. Nevertheless, concentrations in P. amurensis reached as high as 20 microg Se per g dw in October 1996. The enriched concentrations in bivalves (6-20 microg Se per g dw) were widespread throughout North San Francisco Bay in October 1995 and October 1996. Concentrations varied seasonally from 5 to 20 microg Se per g dw, and were highest during the periods of lowest river inflows and lowest after extended high river inflows. Transplanted bivalves (oysters, mussels or clams) were not effective indicators of either the degree of Se contamination in P. amurensis or the seasonal increases in contamination in the resident benthos. Se is a potent environmental toxin that threatens higher trophic level species because of its reproductive toxicity and efficient food web transfer. Bivalves concentrate selenium effectively because they bioaccumulate the element strongly and lose it slowly; and they are a direct link in the exposure of predaceous benthivore species. Biological invasions of estuaries are increasing worldwide. Changes in ecological structure and function are well known in response to invasions. This study shows that changes in processes such as cycling and effects of contaminants can accompany such invasions. PMID- 11879939 TI - Selenium biotransformations into proteinaceous forms by foodweb organisms of selenium-laden drainage waters in California. AB - Selenium contamination represents one of the few clear cases where environmental pollution has led to devastation of wildlife populations, most notably in agricultural drainage evaporation and power plant coal-fly ash receiving ponds. Complex biogeochemistry, in particular extensive biotransformations and foodchain transfer, governs Se ecotoxicology and toxicology, for which the mechanism(s) are still elusive. However, total waterborne Se concentration has been widely used as a criterion for regulating and mitigating Se risk in aquatic ecosystems, which does not account for Se biogeochemistry and its site-dependence. There is a need for more reliable indicator(s) that encompass Se ecotoxicity and/or toxicity. Selenomethionine warrants special attention since it simulates Se toxicosis of wildlife in laboratory feeding studies. While low in free selenomethionine, microphytes isolated from Se-laden agricultural evaporation ponds were abundant in proteinaceous selenomethionine. This prompted a more extensive survey of Se speciation in foodchain organisms including microphytes, macroinvertebrates, fish, and bird embryos residing mainly in the agricultural drainage systems of the San Joaquin Valley, California. Total Se in biomass, water-soluble fractions, and protein-rich fractions were measured along with GC-MS analysis of proteinaceous selenomethionine. In all foodchain organisms, water-soluble Se constituted the major fraction of total biomass Se, while proteinaceous Se was a substantial, if not dominant, fraction of the water-soluble Se. In turn, proteinaceous selenomethionine comprised an important fraction of proteinaceous Se. In terms of total biomass Se, an average 1400-fold of Se biomagnification from water to microphytes was observed while subsequent transfer from microphytes to macroinvertebrates exhibited an average of only 1.9-fold. The latter transfer was more consistent and greater in extent for proteinaceous Se and proteinaceous selenomethionine, which is consistent with their importance in foodchain transfer. Proteinaceous Se in the omnivorous carp (Cyprinus carpio) liver also demonstrated a relation to ovarian lesions, while deformed stilt (Himantopus mexicanus) embryo was more abundant in proteinaceous selenomethionine than were normal embryos. Although limited in the number of organisms surveyed, these findings provide an impetus for further field and laboratory feeding studies to substantiate the hypothesis that proteinaceous selenomethionine underlies Se ecotoxicity, which may in turn prove to be a reliable indicator of Se risk in aquatic ecosystems. PMID- 11879940 TI - Rationale for a tissue-based selenium criterion for aquatic life. AB - This paper proposes a national tissue-based criterion for the protection of aquatic life in the United States based on the growing body of selenium literature, but may be of international importance due to the raising global awareness of selenium contamination. A recent peer consultation workshop was undertaken by the US Environmental Protection Agency to address the technical issues underlying the freshwater aquatic life chronic criterion for selenium. The workshop participants discussed concerns associated with three possibilities for a new criterion: a water-based criterion, a tissue-based criterion, and a sediment-based criterion. Since the current national water quality criterion was established in 1987, several publications have reported adverse effects in fish from dietary selenium exposure with waterborne concentrations below the current criterion of 5 microg/l. Based on this literature, a water-based criterion seems unsuitable because of the propensity for selenium to bioaccumulate through the food chain to toxic dietary concentrations. There is little information to support a sediment-based criterion. A tissue-based criterion accounts for selenium's biogeochemical pathways because it integrates the route, duration, and magnitude of exposure, chemical form, metabolic transformations, and modifying biotic and abiotic factors. The convergence of laboratory and field data shows 4 microg/g to be a conservative value for a national tissue-based criterion for selenium. PMID- 11879941 TI - Development of aquatic life criteria for selenium: a regulatory perspective on critical issues and research needs. AB - The US is currently in the process of revising its freshwater, chronic aquatic life criterion for selenium. The fundamental issues being addressed include which environmental compartment(s) support the most reliable expression of the criterion, which form(s) of selenium should be measured in the medium (media) of choice, and which site-specific water quality (or other factors) should be linked to the expression of the criterion. Literature reviews and a recent workshop were conducted to assess the state of the science on various issues related to water-, tissue- and sediment-based criteria for selenium. Evaluation of many of these issues is ongoing. In terms of water column criteria issues, data limitations will likely restrict the expression of a criterion to operationally defined forms (e.g. total recoverable, dissolved). The specific identity of organoselenium in natural systems is lacking and may not be appropriately represented by free seleno-amino acids (e.g. selenomethionine). The available data do not appear to support quantitative relationships between chronic toxicity and water quality characteristics. In terms of a tissue-based criterion, reproductive tissue (ovary, egg) has been recommended as the tissue of choice, but practical concerns and data availability require consideration of other tissues (e.g. whole-body). Organoselenium (bound to peptides or proteins) is thought to be the form of greatest toxicological importance in fish, however, direct measurements of organoselenium compounds in tissues are very limited. Route of exposure (food vs. water uptake) may prove important for establishing diagnostic tissue residues for selenium based on laboratory data. Data on toxicological aspects of selenium in sediments appear sparse, particularly in relation to different sedimentary forms. Reliable assessments of bioaccumulation will likely be critical for making site specific modifications to chronic selenium criteria, however, many technical issues for assessing bioaccumulation remain. The need for improved analytical methods for directly speciating organoselenium in various environmental media underpins many of the current data gaps. Improving analytical methodologies to enable affordable and reliable measurement of organoselenium compounds holds significant promise for advancing selenium ecotoxicological research. PMID- 11879942 TI - Use of explicit criteria for total hip joint replacement fixation techniques. AB - We developed a tool to judge the appropriateness of indications and fixation mechanisms for total hip joint replacement (THJR) and applied it to a sample of patients. Criteria were developed using a modified Delphi panel judgment process, following the RAND methodology (RAM). We recruited, during 1 year, patients with a diagnosis of osteoarthritis undergoing THJR in five public hospitals. The appropriateness of the THJR intervention and the fixation mechanism was judged by the explicit criteria developed by a panel of experts. Of the 216 scenarios scored by the panel for the use of each of three fixation mechanisms, the cemented fixation was considered inappropriate in 69.5%, versus just 33.3% for the non cemented. Of those scenarios considered appropriate, the most appropriate mechanism of fixation was considered to be non cemented (74.4%), while cemented (17.8%) and hybrid (7.8%) scenarios were scored as appropriate less often. The previous explicit criteria were applied to 583 real patients. After evaluation of the interventions, 30% of the fixation mechanisms used were considered appropriate, while 21.8% inappropriate. Appropriate use of fixation mechanisms varied among hospitals. RAM can provide explicit criteria to help in clinical decision making and evaluating indications for a THJR intervention. Nevertheless, in the case of the appropriateness of fixation mechanisms, due to the lack of evidence, the panel criteria were biased towards the non cemented technique, which had important implications for the evaluation of some hospitals. PMID- 11879943 TI - The evaluation of the introduction of a quality management system: a process oriented case study in a large rehabilitation hospital. AB - OBJECTIVES: So far, there is limited proof concerning the effects of the introduction of quality management systems (QMS) on organisational level. This study concerns the introduction of a QMS in a large rehabilitation hospital. METHODS: Using an observational framework, a process-analysis is performed. The effects were analysed with repeated analyses using the Dutch version of the EFQM model. RESULTS: The introduction of a QMS can be seen as a change process; the pre-change diagnosis proved to be essential. Although many change-related aspects are vital, training and communication, in particular, seemed to be underestimated. Outcomes are a positive correlation between participation in quality activities and work satisfaction and a repeatedly favourable EFQM-score (compared to national levels). CONCLUSIONS: Through a process-analysis, information could be generated to guide organisations in introducing a QMS. An outcome analysis revealed positive effects both in the EFQM-score and the staff's work satisfaction. PMID- 11879944 TI - Setting priorities in Canadian regional health authorities: a survey of key decision makers. AB - INTRODUCTION: The aims of this study were, within three Canadian health regions, to identify existing resource management techniques, to elicit ways in which the current process of setting priorities could be improved and to determine if an economic framework, used internationally, would have merit in Canada. METHODS: Structured, face-to-face interviews were conducted with 62 of 73 key decision makers. This sample included all senior executives and medical directors involved in setting priorities. Descriptive statistics and content analysis were utilised. RESULTS: Key decision makers reported that a clear process of setting priorities does not exist. Allocation of resources generally occurs on the basis of historical trends; only 22% of participants stated that the process works well. Respondents were critical of the lack of transparency and a lack of meaningful inclusion of physicians in the priority setting process. Overall, 92% of respondents indicated that program budgeting and marginal analysis (PBMA) would be an appropriate and useful priority setting framework. CONCLUSIONS: Given the political and historical influence in the process of priority setting and resource allocation, an evidence-based approach, like PBMA which explicitly attempts to identify ways of maximising health benefit within a limited budget, should have merit in the new regional structure in Canada. PMID- 11879945 TI - Diffusion of innovations: treatment of Alzheimer's disease in Germany. AB - Systematic barriers seem to slow down the market penetration of innovative acethylcholinesterase (AChE) inhibitors in Alzheimer's disease. The goal of our study was to examine the diffusion of AChE inhibitors into the German market in more detail. On the basis of using the ongoing surveillance panel of the Institute of Medical Statistic (IMS) Health, the prescription patterns of 100 physicians (72 general practitioners, 28 neurologists) were examined. In addition, structured telephone interviews with the same 100 physicians were conducted. The interview included the assessment of a hypothetical treatment situation (i.e. physicians were asked what they would prescribe if a close relative of theirs had Alzheimer's disease) as well as qualitative items examining the physicians' attitudes towards AChE inhibitors and the perceived impact on drug budgets. As a major result, the analysis revealed that neurologists prescribed AChE inhibitors to 44.6% of their patients, while general practitioners only treated 9.0% of their patients with AChE inhibitors. The analysis of the qualitative items revealed positive attitudes regarding the safety and efficacy of AChE inhibitors, but negative attitudes regarding the budgetary limitations to prescribing these drugs. A correlation of r=0.21 (P<0.05) was found between the perceived impact on drug budgets and the adoption of AChE inhibitors and a correlation of r=0.32 (P<0.002) was seen between the physician's specialty and the adoption of AChE inhibitors. These data show that, while the AChE inhibitor adoption process has passed the early stages, various barriers slow down the final stages of AChE inhibitor adoption. The drug budget in particular seems to inhibit the adoption of the innovation by the majority of general practitioners. This leads to a more short-term cost control strategy instead of long-term disease management and cost saving approaches. PMID- 11879946 TI - Determinants of PHC productivity and resource utilization: a comparison of public and private physicians in Macedonia. AB - The dominant reform paradigm for developing countries introduces market forces into health care provision to improve quality and efficiency. Yet, there is very little empirical evidence as to how individual physicians respond to such incentives. Using a survey of primary health care providers in the Republic of Macedonia, the effect of privatization on physician workload and resource utilization is examined. The survey of physicians in public and private clinics provides extensive data on physician demographics, practice patterns and capital inputs, with an innovation being a measure of physician skill based on responses to several clinical vignettes. Physician production of patient visits is modeled as a jointly determined process of workload and input utilization. Such a formulation acknowledges the endogeneity of input and output and, more importantly, allows the straightforward estimation of the demand equations for labor and capital inputs. Controlling for physician and practice characteristics, private physicians do exhibit higher productivity and greater capital resource use per patient. Major factors influencing workload and resource use are skill and referral rates, both of which have important implications for designing comprehensive and effective physician incentive systems. PMID- 11879947 TI - Nonlinear acoustics in diagnostic ultrasound. AB - The propagation of ultrasonic waves is nonlinear. Phenomena associated with the propagation of diagnostic ultrasound pulses cannot be predicted using linear assumptions alone. These include a progressive distortion in waveform, the generation of frequency harmonics and acoustic shocks, excess deposition of energy and acoustic saturation. These effects occur most strongly when ultrasound propagates within liquids with comparatively low acoustic attenuation, such as water, amniotic fluid or urine. Within soft tissues, similar effects occur, although they are limited by absorption and scattering. Nonlinear effects are of considerable importance during acoustic measurements, especially when these are used to predict in situ exposure. Harmonic generation may be used to create images. These offer improvements over conventional B-mode images in spatial resolution and, more significantly, in the suppression of acoustic clutter and side-lobe artifacts. B/A has promise as a parameter for tissue characterisation, but methods for imaging B/A have shown limited success. PMID- 11879948 TI - Ultrasound as a complement to mammography and breast examination to characterize breast masses. AB - This study was designed to determine if complementary ultrasound (US) imaging and Doppler could decrease the number of biopsies for benign masses. A total of 761 breast masses were sequentially scored on a level of suspicion (LOS) of 1-5, where 1 represented low, and 5 was a high suspicion of malignancy, for mammography, US, and color flow with pulse Doppler (DUS). After biopsy, the results were analyzed using 2 x 2 contingency tables and ROC analysis, for mammography alone and in combination with US and DUS. The addition of US increased the specificity from 51.4% to 66.4% at a prevalence of 31.3% malignancy. ROC analysis showed that the addition of US significantly improved the performance over mammography alone in women < 55 years old (p = 0.049); > 55 years old (p = 0.029); masses < 1 cm (p = 0.016) and masses > 1 cm (p = 0.016). These results show that the addition of US to mammography alone could substantially reduce the number of breast biopsies for benign disease. PMID- 11879949 TI - Ultrasound-guided decompression of the spinal canal in traumatic stenosis. AB - The purpose of this study was to examine the efficacy of ultrasound (US)-guided decompression of the myelon in the surgical treatment of spinal fractures. Intraoperative ultrasonography was performed in 22 patients with traumatic stenosis of the spinal canal during spinal cord surgery with removal of retropulsed bony fragments. US imaging requires a posterior approach and an enlarged foramen interarcuale. The posterior vertebral facet and the myelon can accurately be distinguished from small bony fragments by ultrasonography. Pre- and postoperative computed tomography was compared with intraoperative US imaging. Complete decompression of the spinal canal was controlled by US imaging of the restored ventral epidural space, as seen after repositioning of displaced fragments. Thus, the required extent of the surgical procedure was determined by intraoperative ultrasonography. We conclude that intraoperative US imaging is an important tool to monitor the restoration of the spinal canal and decompression of the spinal cord in case of fracture. The repositioning of stenosing bony fragments using surgical instruments can be monitored. US imaging as a real-time method intraoperatively provides the surgeon with additional information and significantly influences treatment options. PMID- 11879950 TI - Change in retrobulbar circulation during menstrual cycle assessed by Doppler ultrasound. AB - Our purpose was to study the hemodynamic changes in the ophthalmic, central retinal and posterior ciliary arteries during the normal menstrual cycle and to relate the vascular changes to menstrual cycle. A total of 23 healthy women underwent serial color Doppler ultrasonography at least six times during a normal menstrual cycle, twice each in follicular, ovulatory and luteal phases. Pulsatility and resistance index and peak systolic velocity of the each arteries were assessed with color Doppler imaging. There was no statistical difference in any of the parameters during the menstrual cycle. This was supposed to be because generalized hormonal effects on heart rate, blood pressure, blood volume, cardiac output and on the diameter of the vessel cancel each other and this effect maintains the same ocular blood flow and perfusion during the menstrual cycle. PMID- 11879951 TI - Percutaneous assessment of coronary blood flow and cardiac biomarkers. AB - The aim of this study was to compare blood flow determined by coloured microspheres vs. Doppler intravascular ultrasound (US) combined with angiography. A second endpoint was to assess cardiac troponin I (cTnI) as a marker of myocardial injury. Doppler and microspheres were compared in 11 closed chest pigs. Blood flow was measured by catheter-based percutaneous technique in the left circumflex artery (LCx) and compared with coloured microspheres injected in the left ventricle. cTnI was measured in all pigs (73). The mean blood flow (mL/min-1) was 23.3 +/- 8.7 vs. 21.9 +/- 12.1 by Doppler vs. microspheres (p = 0.156), correlation coefficient r = 0.90, p = 0.006. The mean coronary flow with Doppler technique and microspheres in the middle LCx was 22.9 +/- 7.6 vs. 21.2 +/ 6.2 (p = 0.077), and distal 23.9 +/- 10.9 vs. 23.1 +/- 12.1 (p = 0.698). Coronary blood flow measured by Doppler and angiography was comparable to myocardial blood flow measured by coloured microspheres injected in the left atrium or the left ventricle. cTnI was more sensitive to ischaemia than CK-MB mass. PMID- 11879952 TI - Power M-mode Doppler (PMD) for observing cerebral blood flow and tracking emboli. AB - Difficulties in location of transcranial ultrasound (US) windows and blood flow in cerebral vessels, and unambiguous detection of microemboli, have limited expansion of transcranial Doppler US. We developed a new transcranial Doppler modality, power M-mode Doppler (PMD), for addressing these issues. A 2-MHz digital Doppler (Spencer Technologies TCD100M) having 33 sample gates placed with 2-mm spacing was configured to display Doppler signal power, colored red and blue for directionality, in an M-mode format. The spectrogram from a user-selected depth was displayed simultaneously. This system was then explored on healthy subjects and patients presenting with varying cerebrovascular pathology. PMD facilitated window location and alignment of the US beam to view blood flow from multiple vessels simultaneously, without sound or spectral clues. Microemboli appeared as characteristic sloping high-power tracks in the PMD image. Power M mode Doppler is a new paradigm facilitating vessel location, diagnosis, monitoring and microembolus detection. PMID- 11879953 TI - Super harmonic imaging: a new imaging technique for improved contrast detection. AB - For ultrasound contrast agents (UCA), nonlinear imaging now has become fundamental. All of the current contrast-imaging methods are dominantly based on the nonlinear response of UCA bubbles. The discrimination between the perfused tissue and the UCA is the challenge in the field of UCA-imaging. This differentiation is usually associated or expressed by the ratio of the scattered power from the contrast agent to the scattered power from the tissue and is termed "contrast-to-tissue ratio" (CTR). Second harmonic imaging showed a better discrimination between tissue and UCA than fundamental imaging because of a higher CTR. We demonstrate, in this study, that the CTR increases as a function of the order of the harmonic frequency. Currently, due to the limited bandwidth of the transducers, only the second harmonic is selectively imaged, resulting in images with a superior quality to fundamental images, but still degraded and not optimal because of the harmonic generation in the underlying tissue (due to nonlinear propagation) and hence giving a limited CTR. To increase the CTR and to take advantage of the higher harmonics (third, fourth, fifth and the ultraharmonics and termed here super harmonics), we have developed a new phased array transducer. The array transducer contains two different types of elements arranged in an interleaved pattern (odd and even elements). The total number of elements is 96. The elements can operate separately and at a distinct frequency, enabling separate transmission and reception modes. The odd elements (48) operate at typically 2.8 MHz center frequency and 80% bandwidth. The even elements (48) have a center frequency of 900 kHz with a bandwidth of 50%. In vitro measurements using the dual frequency probe show an increase of 40 dB in the CTR for super harmonic components over the conventional second harmonic system. The increase in CTR is in agreement with the calculations using existing models for the response of encapsulated bubbles and known theory of nonlinear propagation. Animal experiments have demonstrated the feasibility of this approach using commercially available UCA and showed a similar increase of the CTR. PMID- 11879954 TI - Movement artefact suppression in blood perfusion measurements using a multifrequency technique. AB - The standard way of suppressing movement artefacts in Doppler measurements is by means of a high-pass filter. This is because artefacts usually are of high amplitude, but have low frequencies. The immediate drawback is, then, that low velocity blood flow is also filtered out. In this paper, a method to reduce movement artefacts in blood perfusion measurements is proposed, using simultaneous transmission and reception of multiple frequencies in a continuous wave Doppler system. It is shown that Doppler signals originating from blood may be considered uncorrelated for a large enough frequency separation between channels, and tissue movements are more correlated. By subtracting perfusion estimates obtained by time-domain processing, correlated signals can be suppressed. The subtraction algorithm is shown to produce a linear perfusion estimate, but with twice the standard deviation compared to an estimate obtained by simply averaging channels. Movement artefacts in both in vitro and in vivo models are shown to be reduced by the algorithm. Imbalance between channels does, however, cause the artefacts to be only partly reduced. The problem can be alleviated by filtering the signals prior to subtraction, but this results in a nonlinear estimate, especially for large time constants in the filter. Some amount of filtering can still be desirable to suppress partly correlated artefacts, even if identical time-domain processing units are implemented, as could be done digitally. PMID- 11879955 TI - Effects of transducer position on backscattered intensity in coronary arteries. AB - Acute myocardial infarction is a frequent cause of sudden death, and is typically initiated by the rupture of coronary artery plaques. The likelihood and severity of rupture are influenced by the plaque structures and components. Radiofrequency (RF) intravascular ultrasound (US) (IVUS-RF) measurements extend current IVUS imaging techniques and may eventually enable the in vivo identification of these features. However, IVUS-RF measurements are affected by the transducer's instantaneous position in the vessel. Specifically, backscattered intensity (BI) decreases as either the distance between the tissue and the transducer increases, or as the beam's angle of incidence on the tissue increases. IVUS-RF data were acquired from seven disease-free coronary arteries in vitro. The 0-dB level for BI was defined as the peak intensity of the reflection from a stainless-steel flat reflector at each distance. The baseline BI measured in adventitial tissue was -32.5 dB (at 0 degrees, 0 mm) with angle and distance dependencies of -0.172 dB/ degrees and -3.37 dB/mm. In contrast, the BI from combined intima and media was -38.2 dB with dependencies of -0.111 dB/ degrees and -4.46 dB/mm (p < 0.05 for all three parameters). Acknowledging and compensating for these effects may allow IVUS-RF to develop into a rapidly deployable tool for the clinical detection of vulnerable plaques and to monitor coronary artery disease progression and regression. PMID- 11879956 TI - New attempt of ultrasonic tissue characterization: decreased chaos in myocardial echo in patients with dilated cardiomyopathy. AB - Ultrasonic radiofrequency (RF) signals returned from the myocardium may display complex behavior, including deterministic chaos, and such chaos may be a useful marker of the tissue properties that are not obtainable with myocardial integrated backscatter. Thus, RF signals were obtained from the myocardium by the transthoracic approach in seven healthy subjects and eight patients with dilated cardiomyopathy (DCM). They were analyzed by a time-delay embedding technique to construct a continuous 3-D trajectory. The attractor formed a thick ring-like structure consisting of a relatively empty, roughly circular, core region in all healthy subjects. In DCM patients, the ring-like structure is thinned with a distinct empty circular core region. When the relation between embedding dimension and averaged correlation dimension was compared, the correlation dimension reached a plateau at the value of 3.5 in healthy subjects, and at the value of 2.6 in DCM patients. In conclusion, chaotic behavior is prevalent in the RF signals for the healthy myocardium, and a decrease in such chaos may be indicative of the damaged myocardium in DCM patients. PMID- 11879957 TI - Axial resolution in elastography. AB - The limits and trade-offs of the axial resolution in elastography were investigated using a controlled simulation study. The axial resolution in elastography was estimated as the distance between the full widths at half maximum of the strain profiles of two equally stiff lesions embedded in a softer homogeneous background. The results show that the upper bound of the axial resolution in elastography is controlled by the physical wave parameters of the ultrasound (US) system used to acquire the data (transducer center frequency and band- width). However, an inappropriate choice of the parameters used to process the US data (cross-correlation window length and shift between consecutive windows) may compromise the best resolution attainable. The measured elastographic axial resolution was found to be on the order of the ultrasonic wavelength. PMID- 11879958 TI - Automated, nonrigid alignment of clinical myocardial contrast echocardiography image sequences: comparison with manual alignment. AB - Analysis of myocardial contrast echocardiography (MCE) images is currently done by manual techniques. The development of computationally efficient methods for aligning images provides an important first step toward the automation of MCE analysis. This is challenging because a nonrigid transformation correction is required. In this paper, we evaluate a state-of-the-art nonrigid alignment method on clinical MCE image sequences (n = 58) acquired on patients during rest and dipyridamole stress, using both B-mode intermittent ultraharmonic (IUH) imaging and real-time myocardial perfusion imaging (RTMPI). Using manual alignment as the reference, we show quantitatively that the automated method aligns images as well as a human observer. However, the new method is faster and more reliable than manual alignment and removes the need for an experienced physician to perform it. The automated technique can be used for quick poststudy off-line analysis and has the potential to be incorporated into an ultrasound machine. PMID- 11879959 TI - Theoretical study on shear stress generated by microstreaming surrounding contrast agents attached to living cells. AB - Numerical calculations have shown that shear stress associated with microstreaming surrounding encapsulated stable bubbles of contrast agents, near living cells driven by 0.12-MPa acoustic pressure amplitude ultrasound (US) at 1 MHz or 2 MHz, may be large enough to generate reparable sonoporation of the cells. Some encapsulated bubbles that have mechanically weak shells may break into free bubbles under the above-mentioned sound field. When that happens, the shear stress caused by microstreaming surrounding the free bubble increases dramatically and may play an important role in lethal sonoporation and fragmentation of cells during the early stage of US exposure. PMID- 11879960 TI - New data on histology and physico-mechanical properties of human tooth tissue obtained with acoustic microscopy. AB - Quantitative evaluation of human tooth structural elements, revealed in acoustic images, has been carried out. It has been shown that tissue elements with different acoustic impedances differed in acoustic images by intensity of grey color, and also feature with different longitudinal sound velocities (C(L)). In the layer of mantle dentin, C(L) is 7% to 8% lower than in bulk dentin, and in the layer of dentin around the pulp chamber, C(L) is 15% lower. In carious enamel and dentin, C(L) decreases up to 7% to 17%. In pathologic teeth, dentin areas with higher density can be revealed; they feature higher C(L); in transparent dentin C(L) can be 15% to 20% higher than in bulk dentin. Results of the present study show that acoustic images reflect internal biomechanical properties of tooth tissue microstructure that can be evaluated quantitatively by means of longitudinal sound velocity determination. PMID- 11879961 TI - International Society for Therapeutic Ultrasound (ISTU). PMID- 11879962 TI - The development of Plasmodium gallinaceum infections in chickens following single infections with three different dose levels. AB - In the present study, groups of 5-day-old chickens were inoculated intravenously with approximately 10(6), 10(4) or 10(2) P. gallinaceum-infected erythrocytes. The outcome of disease in relation to dose level was evaluated in terms of number of parasitized erythrocytes, change in number of erythrocytes, pathological changes of organs and the course of exo-erythrocytic stages of the parasite in various organs over a period of 8 weeks. Mean weight gain and mortality were also recorded. With regard to differences between the size of the inoculation dose and the severity of disease, relationships could be observed in relation to clinical signs, mortality, prepatent period, exo-erythrocytic parasites and severity of pathological changes in organs, but in relation to weight gain, erythrocyte loss and number of parasitized erythrocytes no relationships were observed. PMID- 11879963 TI - The effect of Plasmodium gallinaceum on a challenge infection with Ascaridia galli in chickens. AB - The effect of a primary infection with the haemoparasite Plasmodium gallinaceum on the establishment of a challenge infection with the nematode Ascaridia galli in chickens was studied. Four groups were infected as follows. Group 1: inoculated intravenously with 10(6) P. gallinaceum-infected erythrocytes on day 0; group 2: orally infected with 500 embryonated A. galli eggs on day 10; group 3: infected with P. gallinaceum on day 0 and A. galli on day 10; and group 3: non infected control birds. The results of this investigation demonstrates that a primary infection with P. gallinaceum in chickens alters the course of a subsequent infection with A. galli. Thus, an antagonistic effect was seen in which the malaria infection caused a significant reduction on the establishment of the nematode in concurrently infected animals. PMID- 11879964 TI - Cysteine proteinases Fas1 and Fas2 are diagnostic markers for Fasciola hepatica infection in alpacas (Lama pacos). AB - Circulating antibody against Fasciola hepatica antigens was determined by enzyme linked immunosorbent assay (ELISA) and immunoelectrophoresis in alpacas naturally exposed to F. hepatica. Serological assay parameters were established by using sera from eight infected animals and seven controls with no record of this parasitic infection. Excretory--secretory (ES-) products, Fas1- and Fas2-ELISA were used to survey 307 alpacas from a F. hepatica endemic area in the Peruvian Andes. Seroprevalence of F. hepatica infection varied from 56.7, 64.8 and 66.8% measured by Fas1-, Fas2- and ES-ELISA, respectively. The sensitivity for ES-ELISA was 95%, corresponding Fas1- and Fas2-ELISA sensitivity values were 90 and 95%. In this population, 7% of animals were positive for F. hepatica eggs in faeces, other parasites detected were Trichuris sp. (40%), Nematodirus sp. (34.6%), Lamanema sp. (12.8%) and Eimeria sp. (11.8%). The results show that F. hepatica infected animals elicit circulating antibodies against ES, Fas1 and Fas2. Fas2 ELISA may be proposed as a sensitive assay for the immunodiagnosis of fasciolosis in alpacas. PMID- 11879965 TI - Helminth intensity and diversity in organic meat sheep farms in centre of France. AB - A helminthological study was undertaken at five sheep meat organic farms in the centre of France. The data obtained were compared with two extensive (contemporary of present study) or semi-intensive (literature data) conventional sheep meat farms. The nematode fauna of the region could be characterised by the presence of Trichostrongylus axei alongside with the dominant species recorded in other areas (Teladorsagia circumcincta and Trichostrongylus colubriformis with few Haemonchus contortus). The average intensity of infection was slightly less than 7000 worms in the digestive-tract; infection was on average higher in organic farms, although one organic farm had very low infection. This high intensity could be due to the lower frequency of anthelmintic treatments in these farms. Species/genera diversity was much higher in organic farms. Diversity was positively correlated to the area of pastures available and to the reduction of the number of treatment of ewes. PMID- 11879966 TI - Expression and characterisation of a Psoroptes ovis glutathione S-transferase. AB - The astigmatid mite Psoroptes ovis is the causative agent of sheep scab, a highly contagious parasitic disease of sheep. Infection causes severe allergic dermatitis, resulting in damage to the fleece and hide, loss of condition and occasional mortality. Interest in the P. ovis allergens led us to characterise a glutathione S-transferase (GST) which displays homology to GST allergens isolated from the house dust mite, Dermatophagoides pteronyssinus and the cockroach, Blatella germanica. A cDNA encoding a mu-class GST from P. ovis was expressed in Escherichia coli and the recombinant protein purified for biochemical analysis. SDS-PAGE analysis indicated that the purified product was homogeneous and had an apparent molecular weight of 30 kDa. The recombinant GST (rGST) is active towards the substrate 1-chloro-2,4-dinitrobenzene (CDNB), whereas 1,2-dichloro-4 nitrobenzene (DCNB) is a poor substrate. The recombinant protein was also tested for recognition by IgE and IgG antibodies in serum from P. ovis naive and P. ovis infested sheep. Neither IgE nor IgG antibodies were detected to the rGST. Prausnitz--Kustner testing with rGST did not provoke a characteristic weal and flare response. Biopsies collected at the PK test sites were stained for eosinophils, neutrophils, mast cells and basophils. Neutrophil, mast cell and basophil counts were not significantly different to the controls. Eosinophil numbers were significantly higher than controls, but were not due to an IgE response. PMID- 11879967 TI - Seasonal dynamics of ticks (Acari: Ixodidae) on horses in the state of Sao Paulo, Brazil. AB - Natural tick infestations were assessed every 14 days on horses over a 2-year period. Amblyomma cajennense adult ticks were counted individually, without detachment from the horses. Larvae and nymphs of A. cajennense were collected using a rubber scraper that scratched engorged immature ticks from the host. Adult females of Anocentor nitens larger than 4mm length were counted on the horses. Blood samples were also obtained from the horses every 14 days and macroclimatic data were obtained for the study period. Infestations of A. cajennense demonstrated distinct peaks of activity for each of the three parasitic stages over each 12-month period, showing a 1-year generation pattern. Larvae predominated from April to July and nymphs from June to October. Adults predominated from October to March with a greater number of adult males than females. Although other studies on seasonal dynamics in the states of Rio de Janeiro and Minas Gerais were performed with the free-living stages of A. cajennense on pastures, the present study in the state of Sao Paulo, performed with the parasitic stages of A. cajennense on horses, showed similar results to those observed in other states. Infestations by A. nitens demonstrated distinct peaks of activity of adult females (>4 mm), suggesting different tick generations during the year. Infestation with A. nitens was much higher in the first year than the second year which may have been related to horse nutritional status and stocking rate. Although several climatic variables showed statistical significant correlation (r) with tick counts, the determination coefficients (R(2)) were always lower than 0.40, suggesting that any single significant variable (i.e. mean temperature) would not explain the tick distribution pattern over the year. The highest peaks of A. nitens females (>4 mm) were significantly associated with decrease in horse packed cell volumes (R(2)=0.603). The ears and the perineum, tail and groin region accounted for around 70% of all A. nitens females counted on the horses. PMID- 11879968 TI - The sex ratio of Amblyomma cajennense (Acari: Ixodidae) with notes on the male feeding period in the laboratory. AB - To evaluate the sex ratio of field collected nymphal Amblyomma cajennense ticks, we collected 5326 engorged nymphs from naturally infested horses in Pirassununga county and allowed them to molt to adults in the laboratory. They yielded a sex ratio of 1:1.83 (M:F). Three and two engorged females were collected from horses pastured at Pirassununga county and from tapirs pastured in Sorocaba county, respectively. These females were allowed to oviposit and their progeny were reared until the adult stage in the laboratory. Engorged females collected from Pirassununga yielded a sex ratio of 1:1.57 (M:F) and a sex ratio of 1.14:1 (M:F) were obtained for those ticks collected from tapirs. In addition, unfed tick larvae were collected from Pedreira county and reared in the laboratory until the adult stage. This collection yielded a sex ratio of 1.11:1 (M:F). These results showed significantly different (P<0.05) sex ratio constitutions among different tick populations. Laboratory rabbits were infested once with A. cajennense male ticks, which showed feeding periods varying from 7 to 86 days. During this period, the rabbits were re-infested regularly with A. cajennense female ticks. A total of 179 engorged females were collected from the rabbits and their engorged weight, feeding, preovioposition and egg incubation periods, weight of deposited eggs, percent of hatched eggs and egg production efficiency were compared to the male feeding period and to the number of live males present on the host. None of the female variables were affected by the male feeding period. Male ticks remained fertile for the whole feeding period. Percent of hatched eggs was the only female variable that significantly decreased as the number of live males decreased on the host. The results showed that although some A. cajennense populations are composed of more females than males after molting, this female predominance is compensated by a long male feeding period and maintenance of its reproductive performance. PMID- 11879969 TI - Up-regulated expression of genes encoding Hrk and IL-3R beta subunit by TCDD in vivo and in vitro. AB - Although 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) has been known for its immunosuppressive activity, the mechanisms of its action have been difficult to elucidate, partly because of its inability of exerting its effects in vitro. To gain insights into the molecular mechanisms of immunosuppressive effects of TCDD, we screened for genes, which are regulated by in vivo TCDD treatment in an allogeneic mouse tumor model. RNA, collected from lymphoid organs, was reverse transcribed to cDNA and hybridized to DNA arrays. In addition to genes such as NF kappa B p65 and p27(Kip1) which were previously shown to be regulated by TCDD, expression of several genes including Hrk and IL-3R beta (AIC-2A) was shown to be modulated. RT-PCR and Western blot analyses confirmed the differential expression of Hrk and IL-3R beta. Finally, Hrk was up-regulated by TCDD in Jurkat T cells, suggesting the potential role of Hrk in thymic atrophy and the possibility of exploiting Jurkat T cells as a suitable in vitro model for studying mechanisms of thymic atrophy. PMID- 11879970 TI - The p23 co-chaperone facilitates dioxin receptor signaling in a yeast model system. AB - Hsp90, p23 and other chaperosome proteins are critical for the function of several enzymes and steroid hormone receptors. The dioxin receptor (DR) is an Hsp90-regulated transcription factor that binds numerous toxic ligands, including the environmental contaminant 2,3,7,8-tetrachlorodibenzo(p)dioxin. We used a yeast model system that expressed human DR and Arnt proteins to test whether p23 affected DR signaling. Deletion of the SBA1 gene (yeast p23 homolog) in this model system reduced ligand-mediated DR signaling by approximately 40% and shifted the EC(50) of the beta-napthoflavone ligand by five-fold in a reporter gene assay. DR signaling was restored in the sba1 strain by a plasmid-borne SBA1 gene, confirming that the signaling defect was due to SBA1. The human p23 protein substituted for yeast Sba1 protein in this model system. These genetic data show that p23 enhances DR signaling. PMID- 11879971 TI - Perfluorooctanoate, perflourooctanesulfonate, and N-ethyl perfluorooctanesulfonamido ethanol; peroxisome proliferation and mitochondrial biogenesis. AB - Compounds that cause peroxisome proliferation in rats and mice have been reported to interfere with mitochondrial (mt) bioenergetics and possibly biogenesis. The purpose of this investigation was to establish whether proliferation of peroxisomes and mitochondria are necessarily related. Perfluorooctanesulfonate (PFOS) and N-ethyl perfluorooctanesulfonamido ethanol (N-EtFOSE) were investigated as peroxisome proliferators in comparison to perfluorooctanoic acid (PFOA). Three parameters were chosen to assess peroxisome proliferation, stimulation of lauroyl CoA oxidase activity, reduction of serum cholesterol concentration, and hepatomegaly. mt Biogenesis was assessed through cytochrome oxidase activity, cytochrome content and mitochondrial DNA (mtDNA) copy number. PFOA, PFOS, or N-EtFOSE was administered via a single i.p. injection at 100 mg/kg in male rats, and measurements were made 3 days later. In this model, PFOS and PFOA share similar potencies as peroxisome proliferators, whereas N-EtFOSE showed no activity. mt Endpoints were altered only in the PFOA treatment group, which consisted of a decrease cytochrome oxidase activity in liver tissue and an increase in the mtDNA copy number. None of the perfluorooctanoates significantly altered mt cytochrome content following acute in vivo treatment. These data demonstrate that acute administration of PFOS or PFOA causes hepatic peroxisome proliferation in rats. However, stimulation of mt biogenesis is not a characteristic response of all peroxisome proliferators. PMID- 11879972 TI - Modulation of neurotoxic behavior in F-344 rats by temporal disposition of benzo(a)pyrene. AB - The behavioral changes caused by benzo(a)pyrene (BaP), a polycyclic aromatic hydrocarbon (PAH) compound, were monitored, and also its metabolite levels in cerebellum and cortex were measured in BaP treated rats to see if any relationship existed between these two aspects. Rats were administered 0, 25, 50, 100, and 200 mg/kg of BaP in peanut oil by oral gavage. Plasma, and brain tissue (cerebellum and cortex) samples were collected at 0, 2, 4, 6, 12, 24, 48, 72 and 96 h post administration. Neurotoxic effects peaked at 2 h after dosing and lasted 48 h after dosing for all dose groups. The metabolite levels remained the same from 2 to 4 h, reached a peak at 6 h post gavage and showed a gradual decline returning to baseline levels at 72 h when the motor activity of treatment groups also returned to control levels, indicating recovery from the effects of BaP. A significant (P<0.05) correlation between neurotoxic effects and BaP plasma, and brain metabolite concentrations suggests that metabolism plays an important role in modulating the neurobehavioral effects of BaP. PMID- 11879973 TI - Unique insecticide specificity of human homomeric rho 1 GABA(C) receptor. AB - Several convulsants and major insecticides block the gamma-aminobutyric acid (GABA)-gated chloride channel in brain on binding to the GABA(A) receptor. The GABA(C) receptor, important in retina and present in brain, is also coupled to a chloride channel and is therefore a potential target for toxicant action examined here in radioligand binding and electrophysiological experiments. Human homomeric rho 1 GABA(C) receptor expressed in human embryonic kidney cells (HEK293) undergoes specific and saturable high-affinity binding of 4-n-[3H]propyl-4' ethynylbicycloorthobenzoate ([3H]EBOB) using a cyano analog (CNBOB) to determine non-specific binding. This GABA(C) rho 1 receptor is very sensitive to CNBOB and lindane relative to alpha-endosulfan, tert-butylbicyclophosphorothionate, picrotoxinin and fipronil (IC(50) values of 23, 91, 800, 1080, 4000 and >10000 nM, respectively, in displacing [3H]EBOB). A similar potency sequence (except for picrotoxinin) is observed for inhibition of GABA-induced currents of rho 1 receptor expressed in Xenopus oocytes. The present study does not consider rho 2 homomeric and rho 1 rho 2 heteromeric GABA(C) receptors which are known to be more sensitive than rho 1 to picrotoxinin. The inhibitor sensitivity and specificity of this rho 1 GABA(C) receptor differ greatly from those of human homomeric beta 3 and native GABA(A) receptors. PMID- 11879974 TI - Brominated phenols: characterization of estrogen-like activity in the human breast cancer cell-line MCF-7. AB - A large number of halogenated phenols are detected in the blood of humans, fish and wild-animals. We have characterized the estrogen-like activity of phenol, 4 bromophenol (4-BP), 2,4-dibromophenol (2,4-DBP), 2,4,6-tribromophenol (2,4,6-TBP) and 4-tert-butylphenol (tert-BP) using the estrogen-dependent human breast cancer cell line MCF-7. 4-BP, 2,4-DBP and 4-tert-BP all bind to the estrogen receptor (ER) with approximately 10,000-fold less affinity than 17 beta-estradiol (17 beta E). 2,4,6-TBP was only able to displace 43% of radiolabelled estrogen when tested at concentrations up to 1 microM, whereas phenol had no affinity for the ER. 4 tert-BP stimulated cell growth and induced estrogen-regulated proteins such as the progesterone receptor (PgR) and pS2. The brominated phenols, however, although binding to the ER, did not stimulate cell growth or increase the levels of the PgR or pS2, or reduce the level of 17 beta-E induced pS2. On the contrary, 4-BP, 2,4-DBP and partly 4-tert-BP reduced 17 beta-E-stimulated cell growth apparently by an ER independent mechanism. PMID- 11879975 TI - Toluene metabolites as biological indicators of exposure. AB - The measurement of exhaled and excreted xenobiotics and their metabolites can provide accurate, non-invasive, and time-flexible measurements of internal dose. We analyzed rates of exhaled (2)H(8)-toluene and excreted urinary metabolites from 33 exposures of men to 50 ppm of (2)H(8)-toluene for 2 h at rest. The total dose was distributed as follows: exhaled (2)H(8)-toluene, 13 +/- 6.2%; (2)H(5) hippuric acid, 75 +/- 6.4%; (2)H(7)-o-cresol, 0.31 +/- 0.22%; (2)H(7)-m-cresol, 0.53 +/- 0.44%; and (2)H(7)-p-cresol, 11 +/- 3.8%. Interindividual variability was assessed using the coefficients of variation for peak exhalation or excretion rates, and fractions of dose excreted: (2)H(8)-toluene, c.v.=60, 47%; (2)H(5) hippuric acid, 29, 8.6%; (2)H(7)-o-cresol, 80, 73%; (2)H(7)-m-cresol, 37, 83%; and (2)H(7)-p-cresol, 38, 34%. Excretion rates of the cresols were stable over the first 5 h post-exposure, and o-cresol was determined to be the best urinary indicator of exposure, given the lower background levels of this isomer. The hippuric acid/cresol rate ratios for the first 5 h post-exposure could be described by single exponential terms, and thus provided a means for estimating time since exposure for any finite toluene duration/exposure combination. PMID- 11879976 TI - MRE-binding transcription factor-1 is activated during endotoxemia: a central role for metallothionein. AB - Endotoxin (LPS) has been established to induce hepatic metallothionein (MT), but the specific role of MT remains unknown. In this study, we examined whether MT can modulate MTF-1 activity during endotoxemia. Treatment with IL-6, the main mediator of MT induction during endotoxemia, enhanced the expression of the MRE(d)-driven reporter gene. MTF-1 DNA-binding activity was increased 16-24 h after LPS administration in wild-type mice, while no such activation was observed in MT-null mice during the same period. The expression of alpha(1)-acid glycoprotein (AGP) mRNA, an RNA regulated by MTF-1, was lower in MT-null than in wild-type mice. Our results suggested that MTF-1 was activated during endotoxemia. MT can act as an activator of MTF-1, and MT can induce MTF-1 targeted gene expression during endotoxemia. PMID- 11879977 TI - Association between total cadmium intake calculated from the cadmium concentration in household rice and mortality among inhabitants of the cadmium polluted Jinzu River basin of Japan. AB - A follow-up survey was conducted to investigate the relationship between total Cd intake and mortality in the Jinzu River basin for 6128 days among 757 inhabitants who ingested household rice. When the subjects were divided into two groups, standardized mortality ratios (SMRs) were 0.94 in men and 1.36 in women for the > or = 2.0 g group and 0.68 in men and 0.29 in women for the <2.0 g group (significant in the women). Cox's hazard ratios for men, women, and men+women for the > or = 2.0 g group relative to those in the <2.0 g group were 1.406, 3.955 and 1.950 (significant in the men+women). Using total Cd intake as a continuous variable, the hazard ratios were 1.045, 1.146 and 1.049 in men, women, and men+women (significant in the women), respectively. In the Jinzu River basin, increased total Cd intake appears to exert an adverse influence on life prognosis. PMID- 11879978 TI - The cytotoxic prodigiosin induces phosphorylation of p38-MAPK but not of SAPK/JNK. AB - Prodigiosin (PG) is a red pigment produced by Serratia marcescens, with cytotoxic and immunosuppressive activity. It induces apoptosis in several cancer cell lines, including Jurkat-T cells. Here we examine the role of two stress stimulated kinase cascades in this induction. Time course experiments using polyclonal antibodies showed that p38-MAPK phosphorylation began at 15 min and lasted for 3 h, whereas JNK was not phosphorylated, although both proteins were present. SB203580, a selective inhibitor of p38-MAPK, blocked its phosphorylation in PG-treated cells. Taken together, these data suggest that the PG induces phosphorylation of p38-MAPK but not of SAPK/JNK and that it increases the expression of both c-jun and c-fos oncoproteins. PMID- 11879979 TI - Preventive effect of melatonin against brain mitochondria DNA damage, lipid peroxidation and seizures induced by kainic acid. AB - The effects of kainic acid on mitochondria DNA (mtDNA) or lipid peroxidation in mice brain and, preventive effects of melatonin against its effects were investigated in vivo. Broad-spectrum limbic and severe sustained seizures were observed in all mice when kainic acid (45 mg/kg) was injected intraperitoneally (ip) to eight mice. These seizures were completely abolished by the simultaneous administration of melatonin (20 mg/kg, ip), a potent scavenger of hydroxyl radical. However, slight limbic seizures or severe sustained seizures were observed when melatonin was injected in animals 30 min before or 15 min after the kainic acid administration. The administration of kainic acid caused damage to mtDNA in brain frontal and middle cortex. These effects were abolished when melatonin was injected in animals 0 or 30 min before, but not 15 min after the kainic acid administration. In vitro or in vivo exposure of kainic acid elicited an increase in lipid peroxidation in a concentration- or dose-dependent manner. The increased lipid peroxidation induced by kainic acid was attenuated by co treatment with melatonin. These results indicate that there may be a positive relationship among seizures, brain mtDNA damages and increased lipid peroxidation. Hence, our present results suggest that the hydroxyl radicals produced by kainic acid cause damage on mtDNA and the increase of lipid peroxidation in brain, leading to severe seizures. These effects were completely prevented by co-treatment with melatonin, a potent scavenger of hydroxyl radicals. PMID- 11879980 TI - Role of glutathione in metabolic degradation of dichloromethane in rats. AB - Dichloromethane (DCM) elimination and carboxyhemoglobin (COHb) generation were examined in adult female SD rats pretreated with a glutathione (GSH) depletor(s). Rats were treated with either buthionine sulfoximine (BSO; 2 mmol/kg, i.p.), diethylmaleate (DEM; 3 mmol/kg, i.p.), phorone (PHO; 1 mmol/kg, i.p.) or BSO plus PHO (BSO; 2 mmol/kg +PHO; 0.5 mmol/kg, i.p.). The hepatic GSH concentration was significantly reduced by each treatment. Decrease in hepatic GSH was maintained at least for 10 h after BSO treatment but recovered rapidly in rats treated with DEM or PHO. The hepatic p-nitrophenol hydroxylase activity was not affected by the GSH depletors at the dose used in this study. Rats were treated with an i.p. injection of DCM (3 mmol/kg) and the concentrations of DCM and the COHb levels in blood were monitored. In rats pretreated with a GSH depletor, the peak COHb level was significantly greater than that of rats treated with DCM only. The peak COHb level attained in each group of rats appeared to be inversely related to the magnitude of reduction in hepatic GSH levels. The half-life of DCM in blood was also increased in rats pretreated with the GSH depletor(s). The results indicate that the GSH-dependent metabolic reaction has an important role in the overall elimination of DCM as well as in the metabolic generation of carbon monoxide (CO) from this solvent. PMID- 11879981 TI - Teratogenic assessment of carbadox in rats. AB - Carbadox was administered by gavage once daily to pregnant rats at doses of 0 (control), 10, 25, 50 or 100 mg/kg on days 8 through 15 of pregnancy. The dams were killed on day 21 and the number of implantation sites, resorptions and live fetuses were counted. A significant dose-related decrease in maternal body weight gains during treatment (days 8 through 15 of pregnancy) occurred at doses of 10 mg/kg and above. There was a dose-related decrease in fetal body weights which was statistically significant at 25 mg/kg and above. This compound showed not only embryolethal but teratogenic effect. Resorption rates were 81.8% at 100 mg/kg, occurring complete resorptions in five dams, compared with 3.4% resorption rate in the control. In fetal examinations, a significant increase in the incidence of external, skeletal and internal malformations occurred at 100 mg/kg, where the surviving fetuses born to dams with 40-93% resorptions had any malformations, short tail; kinky tail; brachygnathia or ectrodactyly. PMID- 11879982 TI - Detection of aromatase inhibitors in vitro using rat ovary microsomes. AB - Inhibition of aromatase activity in vitro is one of the Tier 1 screening assays proposed by the Endocrine Disrupter Screening and Testing Advisory Committee (EDSTAC) for the detection of potential endocrine disrupters. In this report a rat ovarian aromatase inhibition assay has been evaluated using the reference aromatase inhibitors anastrozole, fadrozole, letrozole and CGS 18320B. Rat ovary microsomes were used as the enzyme source, as endocrine disruption studies are most commonly carried out in this species. Aromatase activity was inhibited in vitro by anastrozole, fadrozole, letrozole and CGS 18320B with IC(50)s of 25, 7, 7 and 5 nM, respectively. This assay, therefore, appears to have good sensitivity to aromatase inhibitors and may be useful as a general screening assay and in mechanistic studies. PMID- 11879983 TI - Evaluation of the acute and subacute toxicity of a choleretic phloracetophenone in experimental animals. AB - Toxicity of a choleretic compound, phloracetophenone (2,4,6 trihydroxyacetophenone; THA) was investigated in mice, rats and hamsters. Acute toxicity of THA was observed to be dependent on species and route of administration, but not sex and age. LD(50) values for an acute toxicity of a single i.p. administration to adult male hamsters and mice were 338 and 365 mg/kg BW, respectively. It was significantly increased to 489 mg/kg BW in adult male rats and greatly increased by i.g. route. Subacute toxicity was investigated in adult male mice by giving THA at a doses of 37-300 mg/kg BW/day, i.g. for 30 consecutive days. High doses of THA induced periportal hepatocyte degeneration whereas plasma concentrations of alanine and aspartate aminotransferases, bilirubin, and blood urea nitrogen, and hepatic triglyceride content were only slightly increased. The possible therapeutic effect of the choleretic THA was evaluated in the ethinylestradiol (EE)-induced cholestasis. THA enhanced the hepatic clearance of sulfobromophthalein and decreased the elevated plasma alkaline phosphatase in EE-cholestatic rats to control levels. These results suggested that THA at biologically active choleretic dose had low toxicity, it might be safe for further development as a therapeutic agent for a short period of treatment in cholestasis. PMID- 11879984 TI - In vitro nephrotoxicity induced by chloronitrobenzenes in renal cortical slices from Fischer 344 rats. AB - Chloronitrobenzenes are important chemical intermediates in the manufacture of industrial, agricultural and pharmaceutical agents. Toxicity induced by the various chloronitrobenzene isomers in vivo includes hematotoxicity, immunotoxicity, hepatotoxicity and nephrotoxicity. The purpose of the study was to determine the direct nephrotoxic effects of nitrobenzene and ten chlorinated nitrobenzene derivatives using renal cortical slices as the in vitro model. Renal cortical slices were prepared from kidneys of untreated, male Fischer 344 rats and incubated with nitrobenzene (1.0-5.0 mM), a chloronitrobenzene (0.5-5.0 mM) or vehicle for 2 h. At the end of the 2 h incubation, tissue gluconeogenesis capacity (pyruvate-stimulated gluconeogenesis) and lactate dehydrogenase (LDH) release were determined as measures of cellular function and cytotoxicity. Based on decreased pyruvate-stimulated gluconeogenesis and increased LDH release, the order of decreasing nephrotoxic potential was trichloronitrobenzenes>dichloronitrobenzenes>monochloronitrobenzenes>nitrobenzene Among the mono- and dichloronitrobenzenes, 1-chloro-3-nitrobenzene and 3,4 dichloronitrobenzene were the most potent nephrotoxicants, while the two trichloronitrobenzenes tested exhibited similar nephrotoxic potentials. These results demonstrate that chloronitrobenzenes are directly nephrotoxic in vitro and that increasing the number of chloro groups increases the nephrotoxic potential of the resulting chloronitrobenzene derivative. PMID- 11879985 TI - Effects of per os lead acetate administration on mouse hepatocyte survival. AB - Previously published studies indicate that hepatotoxicity is associated with high blood lead (Pb) levels in animal models and humans. The present investigation evaluated the effects of in vivo Pb exposure via drinking water on mouse hepatocyte survival in vitro when blood Pb concentrations reflected those seen in children in urban and rural settings (2-15 microg/dl). The findings indicated a biphasic dose-response with low concentrations associated with a modest decrease in hepatocyte survival, while at the highest concentration, survival was significantly enhanced (60%). Since these responses were associated with concentrations normally encountered by children, follow-up investigations are warranted. PMID- 11879986 TI - Identifying the mechanism of aquatic toxicity of selected compounds by hydrophobicity and electrophilicity descriptors. AB - The most successful quantitative structure-activity relationships (QSARs) have been developed by separating toxicants by their mechanisms of action (MOAs). However, since the activity of a chemical compound on an organism is dependent upon several physical, chemical and biological factors, among which interactions may also exist, the MOA of a compound is not easily determined. In this study, the use of discriminant analysis and logistic regression in distinguishing between narcotic and reactive compounds was investigated. The discriminating variables included hydrophobicity (log(K(ow))) and electrophilicity descriptors (S(av)(N), E(HOMO), and E(LUMO)). Classification results showed that logistic regression gave a smaller total error rate compared to discriminant analysis. Since the value of the descriptors can be calculated, the classification methods can be used in predictive toxicology. PMID- 11879987 TI - Ortho-substituted polybrominated biphenyls activate respiratory burst in granulocytes from humans. AB - The in vivo consequences of exposure to polychlorinated biphenyls (PCB) have been reported to involve reduced phagocytic function, which could be related to increased susceptibility to infections. Though less abundant in the environment, polybrominated biphenyls (PBB) have similar toxicological properties as PCB. In this respect the effect of different PBBs on human granulocytes was elucidated. Ortho-substituted PBBs activated respiratory burst, measured by the chemiluminescence assay, and elevated intracellular calcium. The most active polybrominated congener 2,2',5-TBB increased chemiluminescence in a concentration dependent manner, and ED(50) was approximately 10 microM. PBBs stimulated elevation of intracellular [Ca(2+)] in human granulocytes. The [Ca(2+)]i was elevated from 50 to 250 nM. The respiratory burst due to stimulation by PBBs was inhibited by U73122, ethanol (1%), wortmannin, and bisindolylmaleimide and by the elimination of extracellular calcium in the same way as shown previously for PCBs, indicating that PBB act by the same mechanisms. PMID- 11879988 TI - The role of ketamine on plasma cocaine pharmacokinetics in rat. AB - Ketamine has gained attention recently because of re-emergence of its abuse especially in combination with cocaine. When more than one drug is present simultaneously, the potential for drug--drug interaction exists, which can be pharmacokinetic, pharmacodynamic or both in nature. The objective of this study was to investigate the effect of ketamine on plasma cocaine pharmacokinetics to assess the role that the kinetic component may play in the interaction of these agents. Moreover, the effect of repetitive administration of ketamine pretreatment on the pharmacokinetics of cocaine was addressed. Male Sprague Dawley rats were treated with cocaine alone (5 mg/kg i.v.), ketamine alone (100 mg/kg by gavage), or ketamine followed by cocaine (the same routes and doses). Blood samples were withdrawn at different time points post-injection and analyzed for determination of cocaine, its metabolites (benzoylecgonine and norcocaine) and ketamine. The results demonstrated that ketamine caused a significant decrease in cocaine's area under the curve (AUC) and elimination half-life while its total clearance was increased. The AUC of benzoylecgonine was increased by 1.5-fold after the combination compared with cocaine alone. However, cocaine did not affect ketamine's pharmacokinetic parameters. In the pretreatment study, ketamine was given orally for 3 days, followed 18 h later by a single i.v. of cocaine. Further enhancement of cocaine metabolism occurred with the appearance of norcocaine. This investigation revealed that ketamine enhances cocaine metabolism and may affect its toxicological profile. PMID- 11879989 TI - Lessons learnt from bringing knowledge-based decision support into routine use. PMID- 11879990 TI - HepatoConsult: a knowledge-based second opinion and documentation system. AB - HepatoConsult is a publicly available knowledge-based second opinion and documentation system aiding in the diagnosis of liver diseases. The positive results of a prospective diagnostic evaluation study encouraged its use in clinical routine, although the available hardware infrastructure was not optimal. The comments of the physicians who used the system confirmed the results of the study and showed that the time for data entering is acceptable and the implicit standardization of terminology and documentation is welcome. Suggestions for improvement included the interface to enter data more easily, the scope to be usable for more patients and the additional capability to generate medical reports from the data. PMID- 11879991 TI - Development and evaluation of VIE-PNN, a knowledge-based system for calculating the parenteral nutrition of newborn infants. AB - Calculating the daily changing composition of parenteral nutrition for small newborn infants is troublesome and time consuming routine work in neonatal intensive care. The task needs expertise and experience and is prone to inherent calculation errors. We designed VIE-PNN (Vienna Expert System for Parenteral Nutrition of Neonates), a knowledge-based system (KBS) in order to reduce daily routine work and calculation errors. VIE-PNN was redesigned several times because the clinicians accepted the system only when it saved time. The most recent version of VIE-PNN uses an Hypertext Markup Language (HTML)-based client-server architecture and is integrated into the intranet of the local patient data management system. Since more than 3 years all parenteral nutrition plans are calculated using VIE-PNN. Evaluating the system's performance and the users contentedness, we compared 50 nutrition plans calculated in parallel using VIE PNN or a hand-held calculator, retrospectively analyzed more than 5000 nutrition plans stored in VIE-PNNs database and evaluated a user questionnaire. Nutrition plans were calculated in a mean time of 2.4 versus 7.1min using VIE-PNN or the hand-held calculator. Errors and omissions in the nutrition plans were detected in 22% versus 56% and errors in the VIE-PNN's plans occurring only with interactively changed values. Reviews of stored plans show that a mean of 4 out of 16 parameters were interactively changed. VIE-PNN was well accepted. Most important reasons for the successful operation of VIE-PNN in the daily routine work were time savings and robustness of the system. PMID- 11879992 TI - Efficiency and safety of chemotherapy plans for children: CATIPO--a nationwide approach. AB - Chemotherapy is an important component of childhood cancer treatment. Due to the intensity of this therapy, an error in calculating the dosage of the cytostatic drugs would have severe consequences. Therefore, a computer-aided therapy planning system in pediatric oncology (CATIPO) was introduced into routine use in approximately 20 clinics across Germany. The system consists of a knowledge acquisition module for acquiring knowledge about chemotherapy protocols and a decision support module for producing a patient-specific therapy plan. The main benefits of the system are the reduction of errors and saving time during the development of a particular therapy plan. CATIPO's success is based on the enormous demand for this kind of decision support and its development in tight cooperation with future users. PMID- 11879993 TI - Integrated decision support in a hospital cancer registry. AB - In this paper we present (a) a shell for integrated knowledge-based functions that is destined to support decision processes of the users of the Giessener Tumordokumentationssystem (GTDS) and (b) some results we obtained during a 6 month observation period at one of the customers of the GTDS. A special characteristic of the provided decision support is the high degree of integration in the underlying information system GTDS, i.e. the functions are triggered by events in the patient database, existing patient data is reused as input for the reasoning process and generated alerts are presented instantly to the end-user. The first routine field of application was supporting registrars to adhere to integrity constraints as defined by the International Agency of Research on Cancer (IARC) during the documentation process. This information is important for the registrars since the checks of the IARC are an accepted standard for data quality in cancer registries. The expected benefit of this application area is less effort in achieving adherence to the specification of the IARC by preventing the costly rectification at a later time. During the last 5 months of the observation period 164 alerts were displayed. About 65% of the assessed alerts were considered to be correct. Especially, the analysis of the incorrect alerts revealed some shortcomings in the knowledge behind some of the integrity constraints of the IARC. The general feedback from the end-users indicate positive user satisfaction. Currently, the shell is in use in six hospital cancer registries. PMID- 11879995 TI - Breaking of scored tablets: a review. AB - The literature was reviewed regarding advantages, problems and performance indicators of score lines. Scored tablets provide dose flexibility, ease of swallowing and may reduce the costs of medication. However, many patients are confronted with scored tablets that are broken unequally and with difficulty, reducing compliance and reliance on the drug. Possibilities to reduce breaking difficulties are breaking instructions, tablet-splitters and breaking in advance. Factors influencing the performance of score lines are shape, size, curvature and thickness of the tablet and the form and deepness of the score line. Performance of score lines can be defined by breaking ease, uniformity of mass of subdivided tablets and loss of mass by the subdivision. For breaking ease, an in-vivo reference test and a routinely applicable in-vitro test need to be established. For the uniformity of mass of subdivided tablets a requirement has recently been set by the European Pharmacopoeia. Loss of mass upon breaking can be limited to not more than 1%. PMID- 11879996 TI - Physical stability of redispersible dry emulsions containing amorphous sucrose. AB - The objective of the present study was to estimate the stability of redispersible dry emulsions containing amorphous sucrose. Dry emulsions were prepared by spray drying liquid o/w-emulsions in a laboratory spray dryer. The effect of hydroxypropyl methylcellulose (HPMC) on the glass transition temperature T(g) of spray dried sucrose-HPMC mixtures, relative to the T(g) of amorphous sucrose, was investigated. For the sucrose-HPMC mixtures the values of T(g) followed the ideal Gordon-Taylor equation up to 30% HPMC. For dry emulsions containing 40% HPMC, 30% lipid and 30% sucrose, the T(g) was increased by 12 degrees C relative to the T(g) of amorphous sucrose. The stability of the dry emulsions was investigated by a conventional stability study and by an enthalpy relaxation study. The measured enthalpy recovery of amorphous sucrose below T(g) was used to calculate molecular relaxation time parameters based on the Williams-Watts equation. The molecular mobility of amorphous sucrose at temperatures 50 degrees C below T(g) was low and negligible with respect to the shelf life stability. It was concluded that the dry emulsions are physically stable with respect to the lifetime of a pharmaceutical product when stored in dry condition and at temperatures up to 28 degrees C. PMID- 11879997 TI - Semisolid SLN dispersions for topical application: influence of formulation and production parameters on viscoelastic properties. AB - Aqueous solid lipid nanoparticle (SLN) dispersions with a high lipid content up to 35% and viscous to semisolid consistency were produced by a high pressure homogenization process. Despite their high lipid content and viscosity these dispersions preserved their colloidal size range. The SLN dispersions were compared to nanoemulsions and microparticle dispersions with regard to particle size, viscoelastic properties and formation of a semisolid gel structure. Viscoelastic measurements including oscillation stress sweep tests and oscillation frequency sweep tests demonstrated that the existence of a solid particle matrix with a particle size in the nanometer range is a prerequisite to form a semisolid dispersion having the appropriate consistency for topical application. Striking differences were observed between solid lipid micro- and nanodispersions of the same composition. Particle size reduction resulted in an 80-fold increase of the elastic modulus. Particle size distribution, the physical state of the dispersed lipid phase and the emulsifier concentration have been identified as further key factors for the viscoelastic properties and gel structure of the lipid nanodispersions. By conducting oscillation measurements it was possible to relate the stability of lipid dispersions to specific rheological parameters therefore providing a sensitive tool in stability assessment. Changing the production process from a 40 ml batch to a 2 l batch turned out to have an influence on the colloidal structures of semisolid SLN dispersions. Consistency increased but particle size and ratio of elastic to viscous properties stayed in the same range. PMID- 11879998 TI - Evaluation of superporous hydrogel (SPH) and SPH composite in porcine intestine ex-vivo: assessment of drug transport, morphology effect, and mechanical fixation to intestinal wall. AB - The objective of this study was to investigate the potential of superporous hydrogel (SPH) and SPH composite (SPHC) polymers to enhance the transport of N alpha-benzoyl-L-arginine ethylester (BAEE) and fluorescein isothiocyanate-dextran 4400 (FD4) across porcine intestinal epithelium ex-vivo, and to study any possible morphological damage to the epithelium by applying these polymers. In addition, the ability of these polymers to attach to the gut wall by mechanical pressure was examined by using a specifically designed centrifuge model. The transport of BAEE and FD4 across the intestinal mucosa was enhanced 2- to 3-fold by applying SPHC polymer in comparison to negative control. No significant morphological damage was observed by applying these polymers inside the intestinal lumen. Moreover, the SPH and SPHC polymers were able to attach mechanically to the intestinal wall by swelling and did not move in the intestinal lumen even when a horizontal force of 13 gms(-2) was applied. In conclusion, these polymers are appropriate vehicles for enhancing the intestinal absorption of peptide and protein drugs. PMID- 11879999 TI - Long-term stability of heat-humidity cured cellulose acetate phthalate coated beads. AB - The objective of this study was to investigate the influence of stability storage conditions on the enteric release of heat-humidity cured cellulose acetate phthalate (CAP) coated beads. Theophylline beads were coated with 25 or 35% diethyl phthalate plasticized CAP dispersion (Aquacoat CPD), and cured at a heat humidity condition (50 degrees C/75% RH) for 24h. The cured beads were then stored in various container/closure systems (open glass containers, sealed glass containers with and without desiccant) and exposed to 40 degrees C/75% RH for 6 months or 25 degrees C/60% RH for 12 months. At accelerated conditions (40 degrees C/75% RH), only beads stored in sealed glass containers with desiccant displayed stable release profiles throughout the exposure period. The beads stored in sealed glass containers without desiccant showed increased theophylline release in acidic media at 2h, and did not maintain enteric resistance at 6 months. The release profiles of beads stored in open containers, directly exposed to 40 degrees C/75% RH, were the least stable. The decrease in enteric protection of the beads stored at these two packaging conditions was correlated to an increased phthalic acid content in the films. At ambient storage conditions (25 degrees C/60% RH), all samples possessed enteric release properties, irrespective of the container/closure system. Beads stored in sealed glass containers with desiccant remained the most stable compared to those at the other two packaging conditions. The results indicated that although humidity significantly contributed to coalescence of CAP coating during the curing process, the optimum packaging condition for the heat-humidity cured CAP coated beads was with desiccant to maintain the chemical stability of the CAP. PMID- 11880000 TI - Critical processing factors affecting rheological behavior of a wax based formulation. AB - The use of a wax-based vehicle is one approach to stabilize a drug which is susceptible to hydrolysis and/or oxidation. The drug used in the study, as a microfine powder, is dispersed in the wax mixture and encapsulated in a soft gelatin capsule. To ensure reproducibility of drug content uniformity and encapsulability of the soft gelatin capsule dosage form, optimal viscosity and lot to lot uniformity of the viscosity of the suspension are required. The objective of the study was to identify the critical processing factors which could affect the rheological behavior of the wax based vehicle. Rheological behavior of the vehicle at temperatures ranging from 15 to 90 degrees C was evaluated using a CSL Rheometer equipped with parallel plates and a shear rate sweep mode, unless otherwise specified. Viscosity vs. temperature profiles of the vehicle were determined using the same conditions at different cooling rates ranging from 1.3 to 20 degrees C per min. Three distinct regions of phase transition of the wax mixture can be seen in the Arrhenius plot: (i) a sol region at temperatures above 50 degrees C, (ii) a transition of gel to sol at temperatures ranging from 30 to 45 degrees C, and (iii) a gel region at temperatures below 30 degrees C. The vehicle in a sol region behaved as a Newtonian fluid, indicating minimal interactions between the hydrocarbon chains of the vehicle. The vehicle in a gel region behaved thixotropic in nature, as indicated by a hysteresis loop. The shear rate had a more pronounced effect on the area of thixotropy than the shear time. The cooling rate had a pronounced effect on the resultant viscosity. At the same applied shear rate, the vehicle which was cooled at a faster rate, may cause a recrystallization of the wax mixture in different crystalline forms, resulting in a higher viscosity than the vehicle cooled at a slower rate. This effect was more pronounced when the shear was applied at a lower rate. The results of this study indicate that shear rate and cooling rate are the critical processing factors in controlling the viscosity of the final product and must be well controlled in the manufacturing procedure. PMID- 11880001 TI - Influence of tableting on the enzymatic activity of different alpha-amylases using various excipients. AB - The purpose of the study was to show the influence of compression pressure on the enzymatic activity of different types of alpha-amylases and to analyze the loss of activity of alpha-amylase in mixtures with different excipients. Following that, the properties of excipients used for tableting enzymes were evaluated. Tablets were produced on an instrumented single punch tableting machine. The pure amylases were tableted with increasing graded compaction pressures. Mixtures were tableted to different maximum relative densities, rho(rel,max). The remaining enzymatic activity of the alpha-amylase in the tablets was determined by the starch iodine reaction. The results show a difference between different types of alpha-amylase depending on their origin and additives. Enzymatic inactivation occurs for the pure materials at all pressures used. It is initiated during and continues after compaction. It can be inhibited by freezing the tablets. Another possibility is to tablet the enzyme in a mixture with excipients, which prevent inactivation by softly embedding the enzyme. One example which even stabilizes alpha-amylase at high volume reduction is kappa-carrageenan. In conclusion, enzymatic inactivation can be markedly reduced when excipients are used for tableting, which require little compaction pressure and are able to release the mechanical stress in the form of expansion. PMID- 11880002 TI - Iminodibenzyl as a novel coupling agent for the spectrophotometric determination of sulfonamide derivatives. AB - A rapid, selective and simple spectrophotometric method for the determination of sulfa-drugs is described. The method is based on the formation of violet colored azo product by the diazotization of sulfonamides, viz. sulfathiazole (SFT), sulfadiazine (SFD), sulfacetamide (SFA), sulfamethoxazole (SFMx), sulfamerazine (SFMr), sulfaguanidine (SFG) and sulfadimidine (SFDd) followed by a coupling reaction with iminodibenzyl in alcohol medium. Absorbance of the resulting violet azo product is measured at 570-580 nm and is stable for 24 h at 27 degrees C. Beer's law is obeyed in the concentration range of 0.05-6.0 microg ml(-1) at the wavelength of maximum absorption. The method is successfully employed for the determination of sulfonamides in various pharmaceutical preparations and common excipients used as additives in pharmaceuticals do not interfere in the proposed method. The method offers the advantages of simplicity, rapidity and sensitivity without the need for extraction or heating. A reaction mechanism is proposed for the formation of the violet azo product. PMID- 11880003 TI - The release behavior of brilliant blue from calcium-alginate gel beads coated by chitosan: the preparation method effect. AB - The aim of this study is to reveal how the release behavior of a model drug (brilliant blue, BB) from chitosan coating calcium-alginate gel beads (CCAGB) was influenced by the preparation methods. The CCAGB were prepared by dropping alginate solution into CaCl(2)/chitosan solution (method 1(a)), or into chitosan solution then gelled by CaCl(2) (method 1(b)), or into CaCl(2) solution then coated by chitosan (method 2). Scanning electron microscopy was used for morphology observation, and elemental analysis was applied to determine the chitosan content bound on calcium-alginate gel beads (CAGB). Compared to CAGB, the dried CCAGB had poorer shape and rougher surface morphology especially in methods 1(a) and (b); moreover, CCAGB was found to be more instable in 0.9% NaCl and serious burst of beads occurred when high concentration of alginate (3.0 and 5.0% w/v) was used. The influence on BB release from the beads by chitosan coating was not only related to the chitosan density on bead surface, but also preparation method and other factors. Under un-dried bead state in method 1(a), the increase of chitosan content prolonged BB release in 0.9% (w/v) NaCl; while in method 2, the increase of chitosan concentration over 0.1% (w/v) (3.0% (w/v) alginate concentration was used) resulted in more serious burst of beads and hence facilitated BB release. Furthermore, in both methods 1(a) and 2, the increase of alginate from 1.5 to 3.0 or 5.0% (w/v) usually resulted in the significant burst of beads and accelerated BB release when 0.3 or 0.5% (w/v) chitosan was used for coating. Drying process greatly influenced BB release profile due to the destroying of alginate-chitosan film. The acceleration of BB release from CCAGB by drying process was more significant in the case of method 1 than of method 2. PMID- 11880004 TI - Stability of drug-carrier emulsions containing phosphatidylcholine mixtures. AB - Lipid emulsion particles containing 10% of medium chain triglycerides were prepared using 2% w/w of a mixture 1:1 w/w of purified soya phosphatidylcholine and 2-hexanoyl phosphatidylcholine as emulsifier mixture, for use as drug carriers. The mean droplet sizes of emulsions, prepared using an Ultra Turrax or a high-pressure homogenizer, were about 288 and 158 nm, respectively, compared with 380 and 268 nm for emulsions containing lecithin, or 325 and 240 nm for those containing 6-phosphatidylcholine. The stability of the emulsions, determined by monitoring the decrease of a lipophilic marker at a specified level within the emulsion, and observing coalescence over time, was also greatly increased using the emulsifier mixture. The emulsion stability did not notably change in the presence of a model destabilizing drug, indomethacin. The use of a second hydrophilic surfactant to adjust the packing properties of the lecithin at the oil-water interface provided an increase in the stability of lipid emulsions, and this may be of importance in the formulation of drug delivery systems. PMID- 11880005 TI - Interaction of melatonin with model membranes and possible implications in its photoprotective activity. AB - It is well known that administration of antioxidants represents a successful strategy for preventing the occurrence and for reducing the severity of UV mediated oxidative damage. Melatonin was recently shown to be an efficacious photoprotective agent. The aim of the present study was to better investigate the interaction of melatonin with model membranes and the possible implications in its photoprotective activity. The antioxidant activity of melatonin was tested in two 'in vitro' experimental models: (a) UV radiation-induced peroxidation in phosphatidylcholine multilamellar vesicles (MLVs); (b) scavenging activity against nitric oxide (NO). Furthermore, we investigated the melatonin/biomembrane interaction by differential scanning calorimetry (DSC) on dimyristoylphosphatidylcholine (DMPC) MLVs and unilamellar vesicles (LUVs). The findings of in vitro antioxidant tests suggest that the photoprotective effect of melatonin should be due, partially at least, to the drug scavenging activity against aqueous and lipophilic free radicals, including NO; besides, melatonin might provide its protective effect against UV radiation-induced damage also by acting as a UV-absorbing screen. The results of DSC experiments have evidenced a good capability of melatonin to interact with DMPC bilayers, causing a significant fluidifying effect; however, the transfer of melatonin in the LUVs is faster than that observed for MLVs, even if both values tend to the maximum values reachable. Our present data allow us to emphasize two points: (1) the fluidifying effect induced by melatonin on lipidic bilayers might act as a cooperative mechanism in its protective effect against peroxidative membrane damage; (2) melatonin appears able to cross biomembranes, so that it could protect intracellular components against peroxidative insult. PMID- 11880006 TI - An improved in vitro method for the evaluation of antacids with in vivo relevance. AB - An improved in vitro method for the evaluation of antacids for use with standard equipment is described. The method is a modification of an older method (RIGO method) and has in vivo relevance. The improved method uses USP dissolution test apparatus 2 with a stirring rate of 125 rpm in combination with a computerized automatic burette. The test solution is 250 ml 0.02 M HCl. A total of 20 min after addition of an antacid to the test solution titration starts at a constant speed of 2.0 ml/min 0.1 M HCl. The proposed acceptance criteria for a waiver for clinical studies are: pH after 4 min not less than 2.5 to ensure a rapid onset of effect, pH after 20 min not exceeding 7.0 to ensure that the pH in the stomach remains within physiological values, buffering capacity between pH 2.5 and 4.5 not less than 8 meq/dose and neutralizing capacity not less than 10 meq/dose to ensure sufficient efficacy within the physiological range. The improved method has been validated with respect to robustness to variations in sample preparation, repeatability and intermediate precision and has been cross validated versus the RIGO method. The improved method has been found to be rather insensitive to variations in sample pretreatment and at least equivalent to the RIGO method. PMID- 11880007 TI - Influence of layer position on in vitro and in vivo release of levodopa methyl ester and carbidopa from three-layer matrix tablets. AB - A versatile oral controlled release system for the simultaneous delivery of levodopa methyl ester and carbidopa, consisting of a three-layer matrix tablet, has been studied and developed. Each individual layer of the matrix exhibited a different release mechanism, i.e. the first layer was swellable (S), the second one was erodible (E) and the third one was disintegrating (D). The three layers have been assembled in the monolithic matrix in different relative positions. It was found that in the monolith the three layers could interact, producing in vitro release profiles depending on their relative position. The monoliths having the configurations DSE and SDE were administered to human volunteers in order to determine the plasma profiles. The pharmacokinetic data showed a significant difference between the early time plasma curves: the monolith DSE, having the fast release profile, gave rise to a rapid appearance of a high levodopa plasma level, whereas the slower releasing monolith SDE produced a smoothed plasma concentration profile. PMID- 11880008 TI - Formulation and in vivo evaluation of chlorhexidine buccal tablets prepared using drug-loaded chitosan microspheres. AB - This investigation deals with the development of buccal formulations (tablets) based on chitosan microspheres containing chlorhexidine diacetate. The microparticles were prepared by a spray-drying technique, their morphological characteristics were studied by scanning electron microscopy and the in vitro release behaviour was investigated in pH 7.0 USP buffer. Chlorhexidine in the chitosan microspheres dissolves more quickly in vitro than does chlorhexidine powder. The anti-microbial activity of the microparticles was investigated as minimum inhibitory concentration, minimum bacterial concentration and killing time. The loading of chlorhexidine into chitosan is able to maintain or improve the anti-microbial activity of the drug. The improvement is particularly high against Candida albicans. This is important for a formulation whose potential use is against buccal infections. Drug-empty microparticles have an anti-microbial activity due to the polymer itself. Buccal tablets were prepared by direct compression of the microparticles with mannitol alone or with sodium alginate. After their in vivo administration the determination of chlorhexidine in saliva showed the capacity of these formulations to give a prolonged release of the drug in the buccal cavity. PMID- 11880009 TI - Relation between dissolution profiles and toxicity of cisplatin-loaded microspheres. AB - The aim of this study is to evaluate and compare the dissolution profiles of cisplatin-loaded microspheres (CDDP-MS) in vitro and in vivo, and to determine the relationship between the dissolution profiles in vitro and systemic toxicity. For this purpose, three types of CDDP-MS that release the CDDP for 1, 2 and 5 weeks without a large amount of initial release in phosphate buffered saline (pH 7.4) were prepared. The dissolution profiles of these formulations in vivo were well correlated with in vitro studies, and resulted in well-controlled plasma platinum concentration. The systemic toxicity of the CDDP-MS and CDDP dissolved in saline (CDDP-SOL) were assessed by intraperitoneal administration in mice. The maximal tolerable dose (MTD) of CDDP-SOL was 13.4 mg/kg, whereas the CDDP-MS of 1, 2 and 5-week types were 34.6, 44.2, 62.6 mg/kg, respectively. The MTD of CDDP increased proportionally when 50% of CDDP had been released from MS in vitro (MTD (mg/kg)=5.22 x T(50(day)) + 13.2, R(2)=0.9935). We demonstrate that the systemic toxicity of CDDP-MS can be predicted by evaluation of the dissolution rate in vitro since in vivo dissolution was correlated with the in vitro. PMID- 11880010 TI - Behavior of alginate gel beads containing chitosan salt prepared with water soluble vitamins. AB - Alginate gel beads were prepared which contained weak acid salts of chitosan (Alg CS) and water-soluble vitamins (e.g. ascorbic acid (AS)) and the behavior of the beads, uptake of bile acids was investigated in vitro. The Alg-CS beads rapidly took up bile acid and this phenomenon was observed for both hydrogel beads and dried beads. About 120 micromol of taurocholic acid was taken up into Alg-CS (1 g) prepared with orotic acid. Dried Alg-CS is the granule which can be made easily, and keeps the ability of CS salt, and all elements can be taken as a food. Therefore, Alg-CS could serve as a useful dietary agent for the prevention of hyperlipidemia. PMID- 11880013 TI - General anesthesia versus monitored anesthesia care with remifentanil for assisted reproductive technologies: effect on pregnancy rate. AB - STUDY OBJECTIVES: To compare the outcome of assisted reproductive technology procedures in women who undergo monitored anesthesia care (MAC) with remifentanil versus general anesthesia. DESIGN: Retrospective data analysis. SETTING: University hospital. PATIENTS: 251 ASA physical status I and II women participating in an in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) program. INTERVENTIONS: During the first phase of the study, all patients underwent general anesthesia induction with alfentanil, propofol, and nitrous oxide, which was maintained with isoflurane or propofol infusion. In the second phase of the study, all patients received a standardized MAC technique with a remifentanil infusion; local anesthetics were not used. MEASUREMENTS: The primary endpoint was pregnancy rate per transfer. The number of oocytes collected, fertilized, and cleaved was recorded, as was the number of oocytes transferred. MAIN RESULTS: Patients who underwent MAC had a greater pregnancy rate with IVF (28.2 vs. 16.3%), with ICSI (32.2% vs. 18.8%), and overall (30.6% vs. 17.9%). CONCLUSIONS: Pregnancy rates in women undergoing transvaginal oocyte retrieval for assisted reproductive technologies were significantly higher with a remifentanil-based MAC technique than with a general anesthetic technique. PMID- 11880014 TI - Clonidine premedication prevents preoperative hypokalemia. AB - STUDY OBJECTIVE: To test the hypothesis that clonidine premedication could prevent an increase of plasma epinephrine occurring as a result of anxiety, and a decrease of the serum potassium (K+) levels before the induction of anesthesia. DESIGN: Randomized, double-blinded study. SETTING: University Hospital of Seoul. PATIENTS: 44 ASA physical status I and II patients, aged 20 to 50 years, scheduled for knee, ear, or nose surgery. INTERVENTION: 44 patients were randomly allocated into one of two groups: 22 patients (clonidine group) received clonidine 300 microg orally at 120 minutes before the induction of anesthesia. The other 22 patients (control group) received a placebo. MEASUREMENTS AND MAIN RESULTS: Anxiety level, serum K+, and plasma epinephrine were measured at an outpatient clinic, and immediately before the induction of anesthesia. There were no differences between groups in degree of anxiety experienced, serum K+, or plasma epinephrine levels as measured at the out-patient clinic. Immediately before the induction of anesthesia, the serum K+ levels of the clonidine group were higher than those of the control group (3.89 +/- 0.26 mEq/L vs. 3.50 +/- 0.36 mEq/L), and anxiety and plasma epinephrine levels of clonidine group were lower than those of the control group (p < 0.05). The frequency of hypokalemia (K+ < or = 3.5 mEq/L) of the clonidine group immediately before the induction of anesthesia was significantly lower than that of the control group (0% vs. 50%). CONCLUSIONS: Clonidine premedication was effective in preventing hypokalemic episodes occurring before the induction of anesthesia. PMID- 11880015 TI - Intraoperative use of platelet-plasmapheresis in vascular surgery. AB - STUDY OBJECTIVE: To determine, in a pilot study, whether pheresis of plasma and platelets before surgical blood loss, with reinfusion of the autologous plasma and platelets after completion of the aortic reconstruction, will result in decreased bleeding and decreased transfusion of allogenic blood components in patients undergoing elective aortic reconstruction. DESIGN: Randomized study. SETTING: University medical center. INTERVENTIONS: Patients were randomized to perioperative (acute) platelet plasmapheresis (APP group) versus conventional blood component therapy (control group). In the APP group, blood was withdrawn after induction of anesthesia, to sequester approximately 300 mL of platelet rich plasma (PRP); platelet poor plasma (PPP) and red blood cells (RBC) were sequestered as well. An autotransfusion device was used to collect and re-infuse autologous RBC during the course of the operation in both groups. After completion of the aortic reconstruction, autologous PRP and PPP were re-infused in the APP group. Blood loss, volume of blood component transfusions, and preoperative and postoperative hemoglobin (Hb), hematocrit (Hct), platelet, international normalized ratio (INR), and activated partial thromboplastin time (aPTT) were recorded. MEASUREMENTS AND MAIN RESULTS: There was no difference between groups in demographics, preoperative laboratory values, or surgical procedures, although more patients were treated for aneurysms (73% vs. 60%) and fewer for occlusive disease (20% vs. 40%) in the control versus APP group. Also, there were no differences between the control and APP groups in duration of operation, blood loss, volume of colloid infused, or volume of allogenic RBC and plasma transfused. Patients in the APP group received a greater volume of crystalloid solution (9.1 +/- 3.4 L vs. 6.8 +/- 3.0 L; p = 0.002), but fewer units of allogenic platelets than the control group (0.7 +/- 1.0 units vs. 0.2 +/ 0.4 units; p < 0.04). There were no differences in postoperative Hb, Hct, INR, aPTT, or fibrinogen. The platelet count was lower in the APP group than in the control group (123 +/- 40 x 10(3)/mm(3) vs. 182 +/- 51 x 10(3)/mm(3); p = 0.004). CONCLUSIONS: Perioperative platelet plasmapheresis led to fewer allogenic platelet transfusions in patients undergoing elective aortic reconstruction. However, there was no decrease in blood loss and no reduction in transfusion of allogenic RBC or plasma. Perioperative platelet plasmapheresis is not recommended for routine use in elective aortic reconstruction. PMID- 11880016 TI - The effect of different lidocaine application methods on postoperative cough and sore throat. AB - STUDY OBJECTIVE: To evaluate the efficacy of various ways of lidocaine application in reducing postoperative cough and sore throat. DESIGN: Double blind, randomized study. SETTING: University-affiliated hospital. PATIENTS: 204 ASA physical status I and II patients scheduled for cataract surgery with general anesthesia. INTERVENTIONS: Patients were randomized to six groups (G1, G2, G3, G4, G5, and G6), according to lidocaine application method. Before endotracheal intubation, in the G1 and G2 groups, 10% lidocaine was sprayed on the distal end of the endotracheal tubes (ETTs; G1) and laryngopharyngeal structures (G2). In the G3 group, the distal ends of the ETTs were lubricated with 2% lidocaine jelly. Intravenous (IV) lidocaine was administered to the G4 group at the conclusion of surgery. Intracuff lidocaine was used in the G5 group; in the G6 group, the terminal end of the ETTs were lubricated with normal saline. MEASUREMENTS: At the end of surgery and after extubation, patients were observed to record the number of coughs. At 1 hour and at 24 hours following extubation, sore throat was evaluated. MAIN RESULTS: In the recovery room, 64.4% of the patients experienced cough, with greatest frequency in the G3, G6, and G2 groups, and the least in the G5 and G4 groups. The frequency of sore throat was significantly different among the six groups at 1 hour and at 24 hours, with greater frequency in the G3, G2, and G6 groups. CONCLUSIONS: Using lidocaine to inflate the ETT cuff or IV lidocaine at the end of surgery decreases the frequency of postoperative cough and sore throat and would provide better outcome for patients and the physician. PMID- 11880017 TI - Low-dose dexamethasone reduces nausea and vomiting after epidural morphine: a comparison of metoclopramide with saline. AB - STUDY OBJECTIVE: To compare the efficacy of a low dose of dexamethasone (5 mg) with metoclopramide 10 mg and saline in preventing nausea and vomiting after epidural morphine in posthysterectomy analgesia. DESIGN: Randomized, placebo controlled study. SETTING: Inpatient surgery at Municipal Women's and Children's General Hospital. PATIENTS: 120 ASA physical status I and II women receiving epidural morphine for posthysterectomy analgesia. INTERVENTIONS: All patients received epidural morphine 3 mg for postoperative analgesia. The dexamethasone group (n = 40) received dexamethasone 5 mg, the metoclopramide group (n = 40) received metoclopramide 10 mg, and the saline group (n = 40) received saline. MEASUREMENTS AND MAIN RESULTS: The occurrence of nausea and vomiting appeared more frequently during 6 to 24 hours following the administration of epidural morphine. The total frequency of nausea and vomiting in the dexamethasone group was significantly lower than that of the metoclopramide and saline groups during this period, with reporting frequencies of 21%, 49%, and 53%, respectively (p <.05 each). However, the difference between metoclopramide and saline did not reach statistical significance. CONCLUSIONS: Dexamethasone 5 mg was more effective than metoclopramide or saline in the prevention of nausea and vomiting associated with epidural morphine for postoperative analgesia. PMID- 11880019 TI - Epiglottic hematoma leading to airway obstruction after general anesthesia. AB - Bleeding into the upper airway can cause airway obstruction and death if not recognized promptly. Anesthesiologists are quite familiar with potential airway obstruction from acute epiglottitis, but they may be less familiar with the potential for airway obstruction from epiglottic hematoma. We report what we believe is the second case of epiglottic hematoma after anesthesia and surgery that led to an acute upper airway obstruction. Our case was unique in that there was no excessive airway trauma during tracheal intubation. Most important, this case emphasizes that patients receiving multiple anticoagulants--as our patient was--are at risk for airway bleeding, epiglottic hematoma formation, and airway obstruction. PMID- 11880018 TI - Application of nasal bi-level positive airway pressure to respiratory support during combined epidural-propofol anesthesia. AB - STUDY OBJECTIVE: To examine whether nasal bi-level positive airway pressure (BiPAP) can be used as an airway during combined epidural-propofol anesthesia. DESIGN: Prospective, consecutive case series study. SETTING: Operating room at a general hospital. PATIENTS: 213 ASA physical status I and II adult patients undergoing lower extremity or lower abdominal gynecology surgery. INTERVENTIONS: After epidural anesthesia, propofol was infused at 20 mg/kg/hr (P20) for 4 to 5 minutes followed by 5 mg/kg/hr (P5), and nasal continuous positive airway pressure (CPAP) 8 cm H(2)O and BiPAP 14/8 cm H(2)O was applied. In clinical situations, BiPAP with respiratory rate (RR) 10 breaths/min was applied. Furthermore, tidal volume (V(T)) during anesthesia, the effect of changing pressure support levels, and evaluation of pressure-controlled ventilation without spontaneous breathing were examined. MEASUREMENTS AND MAIN RESULTS: CPAP resulted in a high RR, marked increased PaCO(2), and slightly decreased PaO(2), whereas BiPAP showed no change or a slightly decreased RR, slightly increased PaCO(2), and no change in PaO(2) or a great increase in PaO(2) with oxygen delivery. In clinical applications, similar results were found and anesthetic conditions were sufficient. Tidal volume increased after induction and maintained increased values under BiPAP 14/8 cm H(2)O. Of V(T) at 2, 6, or 10 cm H(2)O of pressure support levels, the 6 cm H(2)O was appropriate. Vecuronium injection showed a slight decrease and then increase in V(T) and PaCO(2), but the values were within normal (safe) limits. Respiration after rapid and high-dose infusion of propofol showed a markedly decreased RR, but the V(T) was maintained, and PaCO(2) and PaO(2) were at safe values. Rapid induction with 2.0 mg/kg propofol followed by P5 showed satisfactory results, in all but the obese patients. CONCLUSIONS: BiPAP 14/8 cm H(2)0 with RR at 10 breaths/min during combined epidural-propofol anesthesia can be used to provide ventilatory support in lower extremity or lower abdominal gynecology surgery. PMID- 11880020 TI - Anesthetic implications of thoracic endometriosis. AB - Endometriosis occurs in 5% to 10% of women of childbearing age and involves the proliferation of endometrial tissue outside the uterine cavity. Thoracic endometriosis is the most frequent extrapelvic manifestation of endometriosis, numbering some 100 reported cases. It may include spontaneous pneumothorax, hemoptysis, chest pain, bronchiectasis, pneumomediastinum, or mediastinal bleeding. Because the tissue is hormonally responsive, all of these manifestations are related to the menstrual cycle (catamenial) and are likeliest to occur during menses. We report the successful anesthetic management of a patient with thoracic endometriosis and recurring catamenial pneumothorax who presented for elective pelvic surgery. PMID- 11880021 TI - Intercostal nerve block with 5% tetracaine for chronic pain syndromes. AB - Treating chronic pain syndromes is always challenging. We describe an effective use of an intercostal nerve block using 5% tetracaine in three patients with postherpetic intercostal neuralgia or postoperative intercostal neuralgia. PMID- 11880022 TI - Epidural blood patch in a patient with an arachnoid cyst. AB - Arachnoid cysts are relatively common occurrences, with the majority being asymptomatic. The safety of an epidural blood patch in a patient with an arachnoid cyst has not been reported. Our patient had a known thoracic arachnoid cyst and required epidural blood patch for a postdural puncture headache. Magnetic resonance imaging obtained following the epidural blood patch demonstrated no alterations of the cyst or spinal cord compression. PMID- 11880023 TI - Uncertainty by choice: anesthesia and the children of night. AB - Anesthesiologists sometimes have difficulty discussing uncertainties with patients. The widespread and deeply visceral uncertainty about sleep, dreams, and death--the daily terrain of the anesthesiologist--has its roots in the classical representations of these states as siblings, the children of the goddess Night. The symbolism of mythology can guide the practitioner by recognizing, through allegory, the range of our own and our patients' fears. PMID- 11880024 TI - Common mechanisms and strategies for prevention and treatment of cardiac arrest during epidural anesthesia. AB - Cardiac arrests continue to occur during epidural anesthesia and frequently result in death or brain damage. Although unintentional "total spinal" anesthesia, respiratory depression, myocardial ischemia, and local anesthetic toxicity can also lead to cardiac arrest, often the arrests do not fit any of these four categories. Many of the unexplained arrests may be attributed to vagal predominance. The evidence for a vagal-linked circulatory mechanism for these arrests is reviewed, and the characteristics that are associated with an increased risk for cardiac arrest during epidural anesthesia are identified. Specific strategies to forestall vagal predominance are discussed. In case these strategies fail, multiple interventions are discussed that should increase the likelihood of a successful resuscitation in the setting of extensive sympathetic blockade. PMID- 11880025 TI - Carcinoid heart disease: a case report and literature review. AB - We report a patient who presented for elective exploratory laparotomy, and resection of a pelvic mass, which was thought to be ovarian carcinoma. Intraoperative transesophageal echocardiography demonstrated right-sided valvular heart lesions, which suggested the diagnosis of carcinoid syndrome before a pathologic confirmation was obtained. This article discusses the classical presentation and anesthetic management of patients with carcinoid syndrome and emphasizes the importance of proper preoperative diagnosis and careful planning if the incidence and severity of the symptoms that this condition can provoke are to be reduced. PMID- 11880026 TI - Anesthesia for endovascular stent repair of a traumatic pseudoaneurysm of the extracranial internal carotid artery. PMID- 11880028 TI - Custom codons come in threes, fours, and fives. AB - In a laboratory coselection of reporter messages containing a single randomized essential two-, four-, five-, or six-base "codon" with suppressor tRNA(Ser) libraries whose members possessed randomized anticodon loops of varying sizes, only four- and five-base "codon-anticodon" interactions survived. These suppressor tRNAs accomplish +1 and -1 frameshift suppression, suggesting biological significance. They also display some properties common to serine tRNAs; such properties include a modest excess of Ser anticodons that might assist tRNA charging. PMID- 11880029 TI - Toward high-throughput synthesis of complex natural product-like compounds in the genomics and proteomics age. AB - In the age of high-throughput biology, novel genes and proteins are emerging quickly. The need for developing organic synthesis-derived methods that allow rapid access to polyfunctional, complex natural product-like compounds is growing constantly, largely because these small-molecule-based compounds serve as smart, powerful tools both in understanding the roles and functions of emerging biological targets and in validating their biological responses. Developing asymmetric synthesis-derived organic reactions on solid phase allows the synthesis of complex natural product-like compounds in a high-throughput manner. Solid phase organic synthesis is now commonly utilized in the library synthesis of rather simple compounds (i.e., compounds with no multiple stereogenic centers). With few exceptions, the synthesis of complex natural product-like derivatives is still in its infancy. Some recent efforts made in this area indicate opportunities yet to be explored. PMID- 11880030 TI - Identification of hNopp140 as a binding partner for doxorubicin with a phage display cloning method. AB - Doxorubicin is a widely used anti-cancer drug. It is assumed to act by inhibiting DNA replication or transcription, although its precise targets and mechanism of cytotoxicity remain unresolved. A T7 phage library expressing human liver cDNA was screened against immobilized doxorubicin to isolate doxorubicin binding proteins. The selected phage contained the C-terminal region of nucleolar phosphoprotein hNopp140, an important factor in the biogenesis of the nucleolus. When the cloned sequence was expressed in E. coli, the recombinant protein was phosphorylated by casein kinase II and oligomerized in the presence of magnesium and fluoride ions, as occurs in vivo. Doxorubicin bound to the expressed protein with a dissociation constant of 4.5 x 10(-6) M, and this interaction was inhibited by the phosphorylation of hNopp140. These results suggested that doxorubicin might disrupt the cellular function of hNopp140. PMID- 11880031 TI - Cell aggregation by scaffolded receptor clusters. AB - The aggregation of cells by lectins or antibodies is important for biotechnological and therapeutic applications. One strategy to augment the avidity and aggregating properties of these mediators is to maximize the number of their ligand binding sites. The valency of lectins and antibodies, however, is limited by their quaternary structures. To overcome this limitation, we explored the use of polymers generated by ring-opening metathesis polymerization (ROMP) as scaffolds to noncovalently assemble multiple copies of a lectin, the tetravalent protein concanavalin A (Con A). We demonstrate that complexes between Con A and multivalent scaffolds aggregate cells of a T cell leukemia line (Jurkat) more effectively than Con A alone. We anticipate that synthetic scaffolds will offer a new means of facilitating processes that rely on cell aggregation, such as pathogen clearance and immune recognition. PMID- 11880032 TI - Conversion of L-proline to pyrrolyl-2-carboxyl-S-PCP during undecylprodigiosin and pyoluteorin biosynthesis. AB - Several medically and agriculturally important natural products contain pyrrole moieties. Precursor labeling studies of some of these natural products have shown that L-proline can serve as the biosynthetic precursor for these moieties, including those found in coumermycin A(1), pyoluteorin, and one of the pyrroles of undecylprodigiosin. This suggests a novel mechanism for pyrrole biosynthesis. The biosynthetic gene clusters for these three natural products each encode proteins homologous to adenylation (A) and peptidyl carrier protein (PCP) domains of nonribosomal peptide synthetases in addition to novel acyl-CoA dehydrogenases. Here we show that the three proteins from the undecylprodigiosin and pyoluteorin biosynthetic pathways are sufficient for the conversion of L-proline to pyrrolyl 2-carboxyl-S-PCP. This establishes a novel mechanism for pyrrole biosynthesis and extends the hypothesis that organisms use A/PCP pairs to partition an amino acid into secondary metabolism. PMID- 11880033 TI - Structure-based computational database screening, in vitro assay, and NMR assessment of compounds that target TAR RNA. AB - There has been little prior effort to discover new drugs on the basis of a unique RNA structure. Binding of the viral transactivator Tat to the 5' bulge of the transactivation response (TAR) element is necessary for HIV-1 replication, so TAR RNA is a superb target. A computational approach was developed to screen a large chemical library for binding to a three-dimensional RNA structure. Scoring function development, flexible ligand docking, and limited target flexibility were essential. From the ranked list of compounds predicted to bind TAR, 43 were assayed for inhibition of the Tat-TAR interaction via electrophoretic mobility shift assays. Eleven compounds (between 0.1 and 1 microM) inhibited the Tat-TAR interaction, and some inhibited Tat transactivation in cells. NMR spectra verified specific binding to the 5' bulge and no interaction with other regions of TAR. PMID- 11880034 TI - A mass spectrometric study of metal binding to osteocalcin. AB - Electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry was used to investigate Ca(2+), Mg(2+), and La(3+) binding to bovine bone osteocalcin (OCN). OCN was shown to bind 3 mol Ca(2+) per mol protein. There was also evidence for the presence of four additional metal binding sites. Ca(2+) increased the formation of the OCN dimer. Mg(2+) bound to OCN to the same extent as Ca(2+) but did not induce the dimerization of OCN. La(3+) bound to a lesser extent than either Ca(2+) or Mg(2+) to OCN and, like Mg(2+), did not influence dimerization. Each Gla residue of OCN participates in Ca(2+) binding, whereas Mg(2+) binding may occur preferentially at sites other than Gla residues. This implies that the different natures of Ca(2+)- and Mg(2+)-containing OCN complexes influence the tendency of OCN to form a dimer. PMID- 11880035 TI - Generation of multiple bioactive macrolides by hybrid modular polyketide synthases in Streptomyces venezuelae. AB - The plasmid-based replacement of the multifunctional protein subunits of the pikromycin PKS in S. venezuelae by the corresponding subunits from heterologous modular PKSs resulted in recombinant strains that produce both 12- and 14 membered ring macrolactones with predicted structural alterations. In all cases, novel macrolactones were produced and further modified by the DesVII glycosyltransferase and PikC hydroxylase, leading to biologically active macrolide structures. These results demonstrate that hybrid PKSs in S. venezuelae can produce a multiplicity of new macrolactones that are modified further by the highly flexible DesVII glycosyltransferase and PikC hydroxylase tailoring enzymes. This work demonstrates the unique capacity of the S. venezuelae pikromycin pathway to expand the toolbox of combinatorial biosynthesis and to accelerate the creation of novel biologically active natural products. PMID- 11880036 TI - Anchor-based design of improved cholera toxin and E. coli heat-labile enterotoxin receptor binding antagonists that display multiple binding modes. AB - The action of cholera toxin and E. coli heat-labile enterotoxin can be inhibited by blocking their binding to the cell-surface receptor GM1. We have used anchor based design to create 15 receptor binding inhibitors that contain the previously characterized inhibitor MNPG as a substructure. In ELISA assays, all 15 compounds exhibited increased potency relative to MNPG. Binding affinities for two compounds, each containing a morpholine ring linked to MNPG via a hydrophobic tail, were characterized by pulsed ultrafiltration (PUF) and isothermal titration calorimetry (ITC). Crystal structures for these compounds bound to toxin B pentamer revealed a conserved binding mode for the MNPG moiety, with multiple binding modes adopted by the attached morpholine derivatives. The observed binding interactions can be exploited in the design of improved toxin binding inhibitors. PMID- 11880037 TI - Glycopeptide biosynthesis in Amycolatopsis mediterranei DSM5908: function of a halogenase and a haloperoxidase/perhydrolase. AB - Glycopeptides are important clinical emergency antibiotics consisting of a glycosylated and chlorinated heptapeptide backbone. The understanding of the biosynthesis is crucial for development of new glycopeptides. With balhimycin as a model system, this work focuses on the investigation of the putative halogenase gene (bhaA) and the putative haloperoxidase/perhydrolase gene (bhp) of the balhimycin biosynthesis gene cluster. An in-frame deletion mutant in the haloperoxidase/perhydrolase gene bhp (OP696) did not produce balhimycin. Feeding experiments revealed that bhp is involved in the biosynthesis of beta hydroxytyrosine, a precursor of balhimycin. A bhaA in-frame deletion mutant (PH4) accumulated glycosylated but nonchlorinated balhimycin variants. The mutants indicated that only the halogenase BhaA is required for chlorination of balhimycin. Nonglycosylated and/or nonhalogenated metabolites can serve as starting points for combinatorial approaches for novel glycopeptides. PMID- 11880038 TI - Exploring the limits of codon and anticodon size. AB - We previously employed a combinatorial approach to identify the most efficient suppressors of four-base codons in E. coli. We have now examined the suppression of two-, three-, four-, five-, and six-base codons with tRNAs containing 6-10 nt in their anticodon loops. We found that the E. coli translational machinery tolerates codons of 3-5 bases and that tRNAs with 6-10 nt anticodon loops can suppress these codons. However, N-length codons were found to prefer N + 4-length anticodon loops. Additionally, sequence preferences, including the requirement of Watson-Crick complementarity to the codon, were evident in the loops. These selections have yielded efficient suppressors of four-base and five-base codons for our ongoing efforts to expand the genetic code. They also highlight some of the parameters that underlie the fidelity of frame maintenance. PMID- 11880039 TI - Transcriptional effects of the potent enediyne anti-cancer agent Calicheamicin gamma(I)(1). AB - We have investigated the mode of action of calicheamicin in living cells by using oligonucleotide microarrays to monitor its effects on gene expression across the entire yeast genome. Transcriptional effects were observed as early as 2 min into drug exposure. Among these effects were the upregulation of two nuclear proteins encoding a Y'-helicase (a subtelomerically encoded protein whose function is to maintain telomeres) and a suppressor of rpc10 and rpb40 mutations (both rpc10 and rpb40 encode RNA polymerase subunits). With longer calicheamicin exposure, genes involved in chromatin arrangement, DNA repair and/or oxidative damage, DNA synthesis and cell cycle checkpoint control as well as other nuclear proteins were all differentially expressed. Additionally, ribosomal proteins and a variety of metabolic, biosynthetic, and stress response genes were also altered in their expression. PMID- 11880040 TI - Selection of v-abl tyrosine kinase substrate sequences from randomized peptide and cellular proteomic libraries using mRNA display. AB - Methodologies for rapidly identifying cellular protein interactions resulting in posttranslational modification of one of the partners are lacking. Here, we select for substrates of the v-abl tyrosine kinase from two protein display libraries in which the protein is covalently linked to its encoding mRNA. Successive selection cycles from a randomized peptide library identified a consensus sequence closely matching that previously reported for the v-abl tyrosine kinase. Selections from a proteomic library derived from cellular mRNA identified several novel targets of v-abl, including a new member of a class of SH2 domain-containing adaptor proteins. Upon modification, several of the substrates obtained in these selections were found to be effective inhibitors of v-abl kinase activity in vitro. These experiments establish a novel method for identifying the substrates of tyrosine kinases from synthetic and cellular protein libraries. PMID- 11880041 TI - Pathway development and pilot library realization in diversity-oriented synthesis: exploring Ferrier and Pauson-Khand reactions on a glycal template. AB - Through a correlation of the ability of small molecules to bind biological macromolecules and their ability to modulate cellular and organismal processes, chemistry can inform biology and vice versa. Diversity-oriented organic synthesis (DOS), which aims to provide structurally complex and diverse small molecules efficiently, can play a key role in such chemical genetic studies. Here we illustrate the trial-and-error experimentation that can refine an initial pathway planning exercise and result eventually in an effective diversity pathway. By exploring Ferrier and Pauson-Khand reactions on a glycal template, we have developed efficient and stereoselective syntheses of tricyclic compounds. In this pathway, diversity results from the substituents and their spatial relationships about the tricyclic rings. A pilot split-pool library synthesis of 2500 tricyclic compounds highlights the use of planning considerations in DOS and a "one-bead, one-stock solution" technology platform. Additionally, it illustrates a promising synthetic pathway for future chemical genetic studies. PMID- 11880042 TI - Cysteine and serine protease inhibitors block intracellular development and disrupt the secretory pathway of Toxoplasma gondii. AB - A number of cysteine and serine protease inhibitors blocked the intracellular growth and replication of Toxoplasma gondii tachyzoites. Most of these inhibitors caused only minor alterations to parasite morphology irrespective of the effects on the host cells. However, three, cathepsin inhibitor III, TPCK and subtilisin inhibitor III, caused extensive swelling of the secretory pathway of the parasite (i.e. the ER, nuclear envelope, and Golgi complex), caused the breakdown of the parasite surface membrane, and disrupted rhoptry formation. The disruption of the secretory pathway is consistent with the post-translational processing of secretory proteins in Toxoplasma, and with the role of proteases in the maturation/activation of secreted proteins in general. Interestingly, while all parasites in an individual vacuole (the clonal progeny of a single invading parasite) were similarly affected, parasites in different vacuoles in the same host cell showed different responses to these inhibitors. Such observations imply that there are major differences in the biochemistry/physiology between tachyzoites within different vacuoles and argue that adverse effects on the host cell are not always responsible for changes in the parasite. Treatment of established parasites also leads to an accumulation of abnormal materials in the parasitophorous vacuole implying that materials deposited into the vacuole normally undergo proteolytic modification or degradation. Despite the often extensive morphological changes, nothing resembling lysosomal bodies was seen in any treated parasites, consistent with previous observations showing that mother cell organelles are not recycled by any form of autophagic-lysosomal degradation, although the question of how the parasite recycles these organelles remains unanswered. PMID- 11880043 TI - Protection of peritoneal macrophages by granulocyte/macrophage colony-stimulating factor (GM-CSF) against dexamethasone suppression of killing of Aspergillus, and the effect of human GM-CSF. AB - Murine peritoneal macrophages in vitro could kill Aspergillus fumigatus conidia, and this activity could be suppressed with dexamethasone. Treatment with granulocyte/macrophage colony-stimulating factor (GM-CSF) alone did not boost killing, but GM-CSF treatment concurrently with dexamethasone reversed the dexamethasone suppression. Both recombinant human and recombinant murine GM-CSF were equivalent in this activity, even though the human reagent reportedly does not stimulate differentiation of murine stem cells. Recombinant human GM-CSF could also reverse dexamethasone suppression of bronchoalveolar macrophage conidiacidal activity. Sequential studies with peritoneal macrophages indicated that recombinant human GM-CSF pretreatment also blocked dexamethasone suppression, but the GM-CSF treatment given after dexamethasone did not block the suppressive effect. Recombinant human GM-CSF did not boost spleen cell proliferation to a mitogenic stimulus, and did not reverse dexamethasone suppression of proliferation. These studies suggest GM-CSF treatment prior to and concurrent with steroid immunosuppression may ameliorate the steroid effect on tissue macrophage antifungal activity, but does not affect steroid suppression of T-cell immunity. PMID- 11880045 TI - Isolation of Nipah virus from Malaysian Island flying-foxes. AB - In late 1998, Nipah virus emerged in peninsular Malaysia and caused fatal disease in domestic pigs and humans and substantial economic loss to the local pig industry. Surveillance of wildlife species during the outbreak showed neutralizing antibodies to Nipah virus mainly in Island flying-foxes (Pteropus hypomelanus) and Malayan flying-foxes (Pteropus vampyrus) but no virus reactive with anti-Nipah virus antibodies was isolated. We adopted a novel approach of collecting urine from these Island flying-foxes and swabs of their partially eaten fruits. Three viral isolates (two from urine and one from a partially eaten fruit swab) that caused Nipah virus-like syncytial cytopathic effect in Vero cells and stained strongly with Nipah- and Hendra-specific antibodies were isolated. Molecular sequencing and analysis of the 11,200-nucleotide fragment representing the beginning of the nucleocapsid gene to the end of the glycoprotein gene of one isolate confirmed the isolate to be Nipah virus with a sequence deviation of five to six nucleotides from Nipah virus isolated from humans. The isolation of Nipah virus from the Island flying-fox corroborates the serological evidence that it is one of the natural hosts of the virus. PMID- 11880044 TI - Cytokines and lymphocyte proliferation in juvenile and adult forms of paracoccidioidomycosis: comparison with infected and non-infected controls. AB - Cellular immune response to Paracoccidiodes brasiliensis antigens (PbAg) was evaluated in patients with the juvenile (JF) and adult (AF) forms of paracoccidioidomycosis as well as in a group of infected individuals living in the endemic area but without any clinical manifestation of the disease. The immune profile of this group of paracoccidioidomycosis-infected individuals was characterized by: 1) a positive skin test to P. brasiliensis antigen; 2) absence of specific antibodies; 3) a vigorous lymphoproliferative response to PbAg; and 4) a typical Th1 pattern of cytokines, with production of IFN-gamma and basal levels of IL-4, IL-5 and IL-10. At the opposite end of the spectrum were the JF patients whose proliferative response to PbAg was significantly impaired and whose cytokine pattern was characteristically Th2, i.e. lower IFN-gamma secretion and significantly higher levels of IL-4, IL-5 and IL-10. These profiles are compatible with forms of higher and lower resistance, respectively. Intermediate immune responses were observed in AF patients, whose specific lymphoproliferative response was lower than in the paracoccidioidomycosis-infected group but higher than in the JF patients. The secretion of IFN-gamma and IL-10 did not differ from the JF group, although IL-4 and IL-5 levels were significantly lower. Since AF patients are able to control fungal dissemination for decades, they can be considered more resistant than JF patients, who manifest the disease soon after infection. PMID- 11880046 TI - Introduction. Antimalarial drug resistance: ramifications, explanations and challenges. PMID- 11880047 TI - Clinical status and implications of antimalarial drug resistance. AB - Africa carries the greatest burden of disease caused by Plasmodium falciparum, and we can expect this burden to rise in the near future, mainly because of drug resistance. Although effective drugs are available (such as artemether lumefantrine, mefloquine, atovaquone-proguanil and halofantrine) they are uniformly too expensive for routine use. Affordable options include chloroquine plus sulfadoxine-pyrimethamine (SP), amodiaquine (alone or in combination with SP) and chlorproguanil-dapsone. Artemisinin combination therapy may offer considerable advantages over alternative therapies, but its introduction faces considerable logistic difficulty. PMID- 11880048 TI - Mechanisms of resistance of Plasmodium falciparum to antimalarial drugs. AB - Chemotherapy and chemoprophylaxis are the principal means of combating malaria parasite infections in the human host. In the last 75 years, since the introduction of synthetic antimalarials, only a small number of compounds have been found suitable for clinical usage, and this limited armoury is now greatly compromised by the spread of drug-resistant parasite strains. Our current knowledge of the molecular mechanisms underlying resistance in the lethal species Plasmodium falciparum is reviewed here. PMID- 11880049 TI - Basis for antifolate action and resistance in malaria. AB - Resistance to antifolates of the malaria parasite Plasmodium falciparum stems from stepwise mutations of the target enzyme dihydrofolate reductase (DHFR). New drugs can be developed against resistant parasites, which are assumed to have limited possibilities in mutations. Mechanisms of resistance other than reduced binding of inhibitors to mutant enzymes may be possible and need to be further explored. New synergistic combinations of drugs targeting DHFR and dihydropteroate synthase may be employed, with new provisions against development of resistance. PMID- 11880051 TI - Emil von Behring and serum therapy. AB - Emil von Behring is the father of serum therapy. We present an overview of the development of this important tool in the treatment of diphtheria. In a historical context Behring's work reflects the scientific spirit of fin de siecle Berlin. PMID- 11880052 TI - Nef: agent of cell subversion. AB - Primate lentiviruses encode a small protein designated Nef that has been shown to be a major determinant of virus pathogenicity. Nef regulates multiple host factors in order to optimize the cellular environment for virus replication. The mechanisms by which this small protein modulates distinct host cell properties provide intriguing insight into the intricate interaction between virus and host. PMID- 11880053 TI - The antimicrobial activity of C-reactive protein. AB - Recent studies in transgenic mice confirmed that C-reactive protein is protective against microbial pathogens. This is consistent with its ability in vitro to bind microbes, activate the complement classical pathway, and engage FcgammaRI and FcgammaRII. However, in transgenic mice protection also requires the alternative pathway of complement, and FcgammaRI is dispensable. PMID- 11880054 TI - Structured treatment interruptions in HIV/AIDS therapy. AB - Structured treatment interruption (STI) has been investigated for three distinct clinical scenarios: during acute infection with the goal of immune reconstitution and auto immunization; during chronic infection, to decrease the amount and toxicity of antiretroviral drugs; and during virologic failure to restore response to subsequent antiretroviral therapy. The potential costs and benefits of STI should be determined. PMID- 11880055 TI - Antibiotic-resistant enterococci: the mechanisms and dynamics of drug introduction and resistance. AB - Enterococci possess a vast array of mechanisms to resist the lethal effects of most antimicrobial drugs currently approved for therapeutic use in humans, thus presenting a considerable therapeutic challenge. This review summarizes current concepts regarding the mechanisms of resistance, as well as the emergence, proliferation, and epidemiology of resistant enterococci. PMID- 11880057 TI - Cholera and climate: revisiting the quantitative evidence. AB - Cholera dynamics in endemic regions display regular seasonal cycles and pronounced interannual variability. We review here the current quantitative evidence for the influence of climate on cholera dynamics with reference to the early literature on the subject. We also briefly review the incipient status of mathematical models for cholera and argue that these models are important for understanding climatic influences in the context of the population dynamics of the disease. A better understanding of disease risk related to the environment should further underscore the need for changing the socioeconomic conditions conducive to cholera. PMID- 11880056 TI - Actinobacillus pleuropneumoniae: pathobiology and pathogenesis of infection. AB - Actinobacillus pleuropneumoniae causes porcine pleuropneumonia, a highly contagious disease for which there is no effective vaccine. This review considers how adhesins, iron-acquisition factors, capsule and lipopolysaccharide, RTX cytotoxins and other potential future vaccine components contribute to colonisation, to avoidance of host clearance mechanisms and to damage of host tissues. PMID- 11880058 TI - Tackling both sides of the host-pathogen equation with Caenorhabditis elegans. AB - If one is interested in dissecting the complex interactions that exist between host and pathogen, the nematode worm Caenorhabditis elegans is perhaps not the first model host that comes to mind. In this review I will introduce 'the worm' and try to show how it is, in fact, well suited to the identification of universal virulence factors and holds great promise for the study of conserved mechanisms of innate immunity. PMID- 11880059 TI - Contributions of plant scientists to the development of the germ theory of disease. AB - Investigations demonstrating the causal role of microbes in diseases of plants were completed decades before Pasteur and Koch presented conclusive evidence in support of the germ theory of disease in man and animals. The limited recognition of these and other contributions in understanding the nature of plant diseases has delayed recognition of the commonality that exists among pathogens in general and mechanisms of pathogenesis. PMID- 11880060 TI - Training the urologic scientist---an endangered species. PMID- 11880061 TI - Review of cadaveric allografts in urology. PMID- 11880062 TI - Efficacy of botulinum-a toxin in children with detrusor hyperreflexia due to myelomeningocele: preliminary results. AB - OBJECTIVES: The established treatment of children with neurogenic bladder consists of the use of anticholinergic drugs, such as oxybutynin and tolterodine, and clean intermittent catheterization four or five times a day. If anticholinergic drugs and clean intermittent catheterization fail, surgery is often necessary. With the intent of avoiding surgery, we investigated the effect of botulinum-A toxin for treating detrusor hyperreflexia in this group of patients. METHODS: The subjects were 17 children (average age 10.8 years) who had detrusor hyperreflexia and were using clean intermittent catheterization four or five times a day. Urodynamic studies were followed by injection of 85 to 300 U of botulinum-A toxin into 30 to 40 sites in the detrusor muscle. Urodynamic follow up was done 2 to 4 weeks after injection. RESULTS: The mean reflex volume increased by 112.1%, from 95.00 plus minus 34.54 mL (range 47 to 147) to 201.45 plus minus 68.57 mL (range 77 to 310) (P <0.005). The maximal bladder capacity increased by 56.5%, from 137.53 plus minus 59.96 mL (range 59 to 242) to 215.25 plus minus 96.36 mL (range 60 to 380) (P <0.005). The maximal detrusor pressure decreased by 32.6%, from 58.94 plus minus 32.32 cm H(2)O (range 19 to 149) to 39.75 plus minus 26.12 cm H(2)O (range 7 to 100) (P <0.005). Detrusor compliance increased by 121.6%, from 20.39 plus minus 26.5 mL/cm H(2)O (range 4.5 to 40) to 45.18 plus minus 45.4 mL/cm H(2)O (range 5.3 to 100) (P <0.01). CONCLUSIONS: Botulinum-A toxin injection into the hyperreflexive detrusor muscle seems to be very effective and might be a therapeutic alternative to anticholinergic drugs. PMID- 11880064 TI - Effect of pentosan polysulfate therapy on intravesical potassium sensitivity. AB - OBJECTIVES: To evaluate further the intravesical potassium sensitivity test (PST) as an indicator of the epithelial leak of interstitial cystitis (IC) and determine whether successful pentosan polysulfate (PPS; Elmiron) treatment is associated with a change in PST results. Most individuals with IC appear to have an abnormally permeable epithelium that allows urinary solutes such as potassium to penetrate to the bladder interstitium, provoking symptoms. METHODS: Data were from an optimal dose trial of PPS in IC. Patients underwent a PST before and after a 32-week trial of 300, 600, or 900 mg PPS/day. The response to PPS treatment was measured using the Patient Overall Rating of Improvement in Symptoms scale. The before and after treatment PSTs and Patient Overall Rating of Improvement of Symptoms scores were compared. RESULTS: Of 377 patients with IC at 28 centers, 302 (80%) had a positive PST at entry. Of the 198 patients who completed the study, 153 were PST positive at entry and 92 (60%) showed clinical improvement at exit. Clinically improved patients had significant improvement on the PST analog pain and urgency scales (3.2 to 1.3 and 3.6 to 1.9, respectively; P <0.0001). In contrast, patients with no clinical improvement had no significant change in pain (3.1 to 2.7) or urgency (3.6 to 3.2). CONCLUSIONS: PST shows abnormal epithelial permeability in most patients with IC and a significant reduction in this permeability after successful PPS therapy. PST appears to be a valid indicator of epithelial abnormality and a reliable test in the diagnosis of IC. PMID- 11880065 TI - Single-dose fluoroquinolone therapy of acute uncomplicated urinary tract infection in women: results from a randomized, double-blind, multicenter trial comparing single-dose to 3-day fluoroquinolone regimens. AB - OBJECTIVES: To compare the efficacy and safety of single-dose and 3-day fluoroquinolone treatment of uncomplicated urinary tract infection (UTI). METHODS: Adult women with acute uncomplicated UTI were randomized to receive either a single dose of gatifloxacin (400 mg), 3 days of gatifloxacin (200 mg daily), or 3 days of ciprofloxacin (100 mg twice daily). Patients were assessed at four points during the study: before treatment (within 48 hours before the initiation of the study medication), at the end of treatment (by telephone contact on day 3), and twice after treatment completion (5 to 9 days after treatment [test-of-cure visit] and 29 to 42 days after treatment [only patients with a bacteriologic response of eradication at the test-of-cure visit]). RESULTS: The bacterial eradication rate for the single-dose gatifloxacin, 3-day gatifloxacin, and 3-day ciprofloxacin groups was 90%, 95%, and 89%, respectively; the clinical efficacy rate was 93%, 95%, and 93%, respectively, for microbiologically assessable patients at the test-of-cure visit. Eradication of the most common uropathogens, including Escherichia coli, Klebsiella pneumoniae, and Proteus mirabilis, was achieved with gatifloxacin and ciprofloxacin. Single dose gatifloxacin was equivalent to 3-day ciprofloxacin in both microbiologic and clinical efficacy. CONCLUSIONS: Single-dose and 3-day gatifloxacin were microbiologically and clinically equivalent to 3-day ciprofloxacin for the treatment of acute UTI among women. Single-dose gatifloxacin may offer advantages over 3-day fluoroquinolone therapy for uncomplicated UTI by decreasing secondary use of medical resources and improving patient compliance. PMID- 11880066 TI - Prospective study on ultrasonography plus plain radiography in predicting residual obstruction after extracorporeal shock wave lithotripsy for ureteral stones. AB - OBJECTIVES: To compare ultrasonography (US) and plain radiography with intravenous urography (IVU) in predicting ureteral obstruction after in situ extracorporeal shock wave lithotripsy (ESWL) for ureteral stones. METHODS: From April 1998 to September 2000, 100 consecutive patients with solitary ureteral stones were treated by primary in situ ESWL. ESWL failures were salvaged by ureteroscopic lithotripsy. Ninety-three patients completed the follow-up assessment. US and IVU were performed when plain radiography showed no residual stone. The occurrence of hydronephrosis on US was compared with IVU, the reference standard for ureteral obstruction. RESULTS: Of the 93 patients, 72 were men and 21 women (mean age 52 years; mean stone size 11.2 mm). ESWL successfully treated 70 ureteral stones (75%), and the 23 failures were treated by ureteroscopic lithotripsy. Sixty-nine patients without hydronephrosis on US had no ureteral obstruction on IVU. Of the 24 patients who had hydronephrosis on US, 8 had ureteral obstruction on IVU. Of the 85 patients who had no ureteral obstruction on IVU, 69 patients showed no evidence of hydronephrosis on US. However, all patients with ureteral obstruction on IVU demonstrated hydronephrosis on US. The sensitivity, specificity, and positive and negative predictive value concerning sonographic hydronephrosis in the prediction of ureteral obstruction was 100%, 81%, 33%, and 100%, respectively. US alone could not define the cause of ureteral obstruction. CONCLUSIONS: Plain abdominal radiography plus US is highly sensitive for screening ureteral obstruction after primary in situ ESWL for ureteral calculi. It can save up to 74% of patients from the potential risk of IVU. The detection of the cause of obstruction by IVU is only necessary when sonographic evidence of hydronephrosis is present. PMID- 11880067 TI - Urolithiasis in allograft kidneys. AB - OBJECTIVES: Allograft stones are an uncommon clinical problem and management is mainly based on anecdotal experience, rather than analysis of larger series. METHODS: In an 8-year period, 19 patients were treated for 19 renal and 3 ureteral stones. In 9 patients, stones were transplanted and 10 formed de novo stones within a mean of 28 months (range 13 to 48) after transplantation. In 4 patients, stones were removed during transplantation. Seven patients were treated with extracorporeal shock wave lithotripsy (ESWL), 3 patients had stones removed percutaneously, 1 by antegrade ureteroscopy, and 1 at the time of ureteral reimplantation. Three patients passed stones spontaneously. RESULTS: In 3 of 4 patients with stones detected before transplantation, the procedure was completed successfully after endoscopic stone removal. Three of 5 patients with transplanted stones required emergency nephrostomy; 1 patient had permanent renal impairment. Three (42.9%) of 7 patients treated with ESWL needed transient nephrostomy; ultimately, all became stone free within a mean 15 days (range 10 to 40). Endoscopic stone removal always resulted in complete clearance without renal impairment. All patients were stone free during a follow-up of 29 months (range 13 to 48). CONCLUSIONS: Nine (47%) of 19 stones were actually transplanted. Therefore, intraoperative screening by ultrasonography with subsequent endoscopic removal is advisable. Small stones (4 mm or less) may be closely followed up, because they can pass spontaneously. ESWL is the treatment of choice for caliceal stones sized 5 to 15 mm. However, for stones greater than 15 mm or for ureteral stones, antegrade endoscopic procedures seem to be more favorable. PMID- 11880068 TI - Use of intravenous anesthesia for tension-free vaginal tape therapy in elderly women with genuine stress incontinence. AB - OBJECTIVES: To study the efficacy of the tension-free vaginal tape (TVT) procedure for elderly patients. METHODS: Forty-five women, 65 years old or older (mean 69.1; range 65 to 85), were treated for genuine stress incontinence with the tension-free vaginal tape procedure under intravenous anesthesia with heavy sedation. Urodynamic studies and 1-hour pad test were performed before the procedure and at 1 year postoperatively. Vigorous manual pressure against the abdominal wall and ensuring no lifting of urethra by intraoperative introital ultrasonography were used to position and adjust the vaginal taping. RESULTS: Forty-one (91%) were cured, one improved, and three failed. No major surgical complications occurred. Two bladder perforations were noted. The pad test disclosed a reduction in urine leakage from a mean of 28.9 to 1.6 g/hr (P <0.001). The average blood loss was 72 mL (range 30 to 250), operating time was 21 minutes (range 18 to 35), and postoperative hospital stay was 24.2 hours (range 12 to 72). Spontaneous voiding with an adequate volume of postvoid residual urine was noted for all patients by the fourth day postoperatively. Urodynamic parameters related to the voiding dysfunction showed no significant difference before and after surgery. De novo detrusor instability was observed in 2 patients. CONCLUSIONS: With modifications of the technique and the aid of intraoperative ultrasonography, the TVT procedure can be performed under intravenous anesthesia with heavy sedation. The procedure is safe, effective, and minimally invasive for elderly patients. However, long-term follow-up is needed to determine whether this surgery achieves long-lasting results. PMID- 11880069 TI - Transposition of the left renal vein for treatment of the nutcracker phenomenon: long-term follow-up. AB - OBJECTIVES: To assess the therapeutic value of left renal vein transposition for treatment of the nutcracker phenomenon in long-term follow-up. METHODS: Eight patients (4 women and 4 men) between 23 and 58 years old (mean 39.1) underwent transposition of the left renal vein for treatment of the nutcracker phenomenon associated with recurrent gross hematuria and flank pain. The postoperative follow-up was 41 to 136 months (mean 66.4). RESULTS: No perioperative complications were encountered. The postoperative complications comprised deep vein thrombosis (n = 1), retroperitoneal hematoma necessitating surgical revision (n = 1), and paralytic ileus that resolved with conservative management (n = 1). One patient underwent laparotomy for treatment of mechanical ileus due to adhesions 4 years after the initial surgery. In 7 of 8 patients, transposition of the left renal vein efficiently relieved the symptoms related to the nutcracker phenomenon. In 1 patient, the hematuria persisted despite postoperative normalization of the pressure gradient between the left renal vein and the inferior vena cava. CONCLUSIONS: Transposition of the left renal vein is an efficient surgical approach for the treatment of the nutcracker phenomenon and is associated with an acceptable risk of complications. However, rare cases may be encountered in which the shunted connections between the renal veins and the collecting system are so matured that, despite removal of the obstruction of the renal venous backflow, gross hematuria may persist. PMID- 11880070 TI - Coloepiploic mobilization during left radical nephrectomy for renal cell carcinoma is indicated to reduce the risk of iatrogenic splenectomy. AB - OBJECTIVES: To determine whether coloepiploic mobilization (CEM) is indicated to reduce the incidence of iatrogenic splenectomy during left radical nephrectomy for renal cell carcinoma. The incidence of iatrogenic splenectomy during a left nephrectomy is estimated to be between 1.4% and 24%. In a recent study, we reported that the incidence of iatrogenic splenectomy was 8% during a left nephrectomy performed for renal cell carcinoma through a transperitoneal anterior subcostal incision. METHODS: A left radical nephrectomy was performed in 233 consecutive patients for renal cell carcinoma through a transperitoneal anterior subcostal incision with a CEM procedure in which the left colonic flexure was completely detached from the epiploa. Perioperative and postoperative complications, including splenic injury, were noted in a database. The mean patient age was 51.3 years (range 21.3 to 90.2). The mean tumor size was 58 mm (range 15 to 230). RESULTS: An iatrogenic splenectomy was required in 3 patients, and in 1 patient, a splenic injury was treated conservatively. The incidence of iatrogenic splenectomy accompanying left radical nephrectomy was 1.3%. The mean operative time was 120 minutes (range 80 to 240). The mean time to normal gut motility was 3.4 days (range 2 to 11) and to discharge from the hospital it was 9.3 days (range 6 to 19). Regarding CEM, we did not observe any significant abdominal complications. CONCLUSIONS: The incidence of iatrogenic splenectomy during a left radical nephrectomy through a transperitoneal anterior subcostal incision may be reduced by performing the technique of CEM. PMID- 11880072 TI - A method for estimating within-patient variability in maximal urinary flow rate adjusted for voided volume. AB - OBJECTIVES: To investigate whether volume adjustment can be used to reduce the within-patient variability of the maximal urinary flow rate (Qmax) without affecting the variability between patients. METHODS: We analyzed 2049 urinary flow curves of 208 men with lower urinary tract symptoms suggestive of bladder outlet obstruction and studied the relation between Qmax and voided volume (VV) in individual patients using multilevel regression analysis. RESULTS: In agreement with most previous studies, we found a hyperbolic relation between the VV and Qmax. Although hyperbolic on average, the slope of the regression line relating the VV to Qmax differed substantially across individual patients. In some patients, Qmax even tended to be lower with an increased VV. CONCLUSIONS: The reproducibility of Qmax is poor, in part because of its dependence on the VV. The current methods to adjust Qmax for VV, such as the Siroky nomogram, minimize between-patient dependency, whereas our findings emphasize the importance of within-patient variability. A common approach to adjust Qmax for VV, however, is not feasible, because the relation of Qmax to VV differs substantially across patients. The only valid, although impractical option, remains to record a large number of urinary flow curves for each patient. PMID- 11880071 TI - Insulin-like growth factor-1, insulin-like growth factor binding protein-3, and body mass index: clinical correlates of prostate volume among Black men. AB - OBJECTIVES: To examine the relationship between insulin-like growth factor-1 (IGF 1), insulin-like growth factor binding protein-3 (IGFBP-3), and body mass index and prostate volume, as a surrogate marker for benign prostatic hyperplasia, in a community-based sample of black men. Epidemiologic studies examining the role of IGF-1 and IGFBP-3 suggest that increased levels of serum IGF-1 and decreased levels of serum IGFBP-3 are associated with an increased risk of prostate cancer. Few studies have examined these factors with respect to benign prostatic hyperplasia, and these have been limited to white men. METHODS: The study population consisted of a sample of 364 black men, 40 to 79 years of age, residing in Genesee County, Michigan. Men with prostate cancer or prior prostate surgery were excluded. All subjects completed a clinical examination, which included a complete urologic examination with transrectal ultrasonography, anthropometric measurements, and serum assays for IGF-1 and IGFBP-3. RESULTS: Multivariable regression models demonstrated that prostate volume increased with increasing age (P <0.0001) and body mass index (P = 0.03). IGFBP-3 rather than IGF-1 was positively associated with increasing prostate volume (P = 0.003). CONCLUSIONS: This is the largest study describing the relationships between IGF 1, IGFBP-3, and body mass index and prostate volume, and the only study in black men. Although earlier studies demonstrated an association between IGF-1 and prostate cancer risk, our findings indicate that IGFBP-3 is more relevant for prostate enlargement, suggesting that IGF-1 and IGFBP-3 may play different pathophysiologic roles in benign and malignant prostatic conditions. PMID- 11880073 TI - Effects of finasteride and cyproterone acetate on hematuria associated with benign prostatic hyperplasia: a prospective, randomized, controlled study. AB - OBJECTIVES: To evaluate the influence of two differently acting antiandrogens, finasteride (FIN) and cyproterone acetate (CPA), on the natural history of hematuria associated with benign prostatic hyperplasia (BPH) in a prospective, randomized, controlled study. METHODS: Forty-two patients with hematuria episodes due to BPH were randomly allocated to three subgroups of 14 patients each and treated daily with either 5 mg FIN or 100 mg CPA or were placed in a watchful waiting arm. Patients were evaluated at 3-month intervals, and 40 patients had at least 1 year of follow-up. RESULTS: Four patients in the FIN group (30%) and three in the CPA group (23%) presented with recurrent hematuria. In both groups, the bleeding episodes were treated conservatively and required no hospitalization. In the control group, 8 patients (57%) presented with recurrent bleeding; in 4, the bleeding was severe and required some form of intervention (catheterization or transurethral prostatectomy). When the frequency and severity of the hematuria episodes were analyzed over time, a statistically significant difference for FIN versus control was present at 9 and 12 months (analysis of variance, P = 0.035 and P = 0.009, respectively). A similar difference was evident for CPA versus control at 9 and 12 months (P = 0.028 and P = 0.008, respectively). No statistically significant difference was present between the FIN and CPA groups. Interestingly, no statistically significant effect in bleeding recurrence for both CPA and FIN over controls was present at 3 and 6 months of follow-up. CONCLUSIONS: Both FIN and CPA seem to exert a comparable control in hematuria recurrence in patients with BPH, thus confirming the rationale behind the use of antiandrogens for such a purpose. Our results support the hypothesis that any antiandrogen, irrespective of the mode of action, would alter the natural history of BPH-associated hematuria. Interestingly, our results indicate that the speed of action of FIN may not be as rapid as previously described. PMID- 11880074 TI - Rise and fall of radical prostatectomy rates from 1989 to 1996. AB - OBJECTIVES: To describe the changes in the rates of radical prostatectomy procedures, prostate-specific antigen (PSA) screening tests among Medicare beneficiaries, and the incidence of prostate cancer in the United States and to explain the exaggerated increase and decrease in the frequency of radical prostatectomy from 1989 to 1996. METHODS: Medicare claims data on radical prostatectomy procedures and screening PSA tests and the National Cancer Institute's Surveillance, Epidemiology, and End Results prostate cancer incidence data were used to estimate the rates of PSA testing and radical prostatectomy among Medicare beneficiaries aged 65 to 74 years and 75 years and older (population rates). The age-specific true rates of the procedure were also estimated for the incident cases of prostate cancer (the population at risk of undergoing radical prostatectomy) among the beneficiaries. RESULTS: The PSA test, prostate cancer incidence, and radical prostatectomy rates increased from 1989 to 1992. Thereafter, the incidence of prostate cancer, and the population and true rates of radical prostatectomy declined. The percentage of increase and decrease in the population rate of radical prostatectomy was approximately twice that in its true rate. CONCLUSIONS: The radical prostatectomy rates based on all Medicare beneficiaries grossly exaggerated the changes in the use of the procedure. Where possible, true rates, using the population at risk as the denominator, should be used in the studies of diagnostic and therapeutic procedures, complications, and adverse effects. PMID- 11880075 TI - Coping with prostate cancer: a quantitative analysis using a new instrument, the centre for clinical excellence in urological research coping with cancer instrument. AB - OBJECTIVES: To assess the reliability (internal and interrater) and validity (concurrent) of a new interview measure for assessing patients' ability to cope with cancer, the Centre for Clinical Excellence in Urological Research Coping with Cancer Instrument (CCCI), and to determine whether there is an underlying structure to the various coping strategies used by patients with prostate cancer. METHODS: Eighty patients with prostate cancer were interviewed using the CCCI. The participants also completed measures of quality of life and anxiety and depression. RESULTS: The psychometric properties of the CCCI were acceptable. Factor analysis revealed that coping with prostate cancer can be described along five dimensions: positive problem solving (fighting against the illness, seeking information); self-reliance (developing a lay explanation, distrusting doctors); emotional availability (not withdrawing from others); distress (brooding, self pity); and solace (taking alcohol or drugs to improve mood). These coping styles were correlated with age, quality of life, self-reported prostate-specific antigen level, and measures of anxiety and depression. CONCLUSIONS: The results of the present study have led to a greater understanding of the underlying coping styles used by patients with prostate cancer. Although some of these are similar to those found in other cancer populations, others, such as self-reliance and solace, represent unique and potentially clinically significant responses to prostate cancer diagnosis and treatment. A larger scale longitudinal study is needed to determine the wider clinical implications associated with each coping style. PMID- 11880076 TI - Concurrent assessment of obstructive/irritative urinary symptoms and incontinence after radical prostatectomy. AB - OBJECTIVES: To concurrently evaluate urinary obstructive symptoms and incontinence in men after radical prostatectomy and to determine the effects of these components on urinary impairment and satisfaction. METHODS: Two hundred twenty-eight men after radical prostatectomy were age-matched and zip code matched to a random sample of 228 men without prostate cancer. Urinary incontinence was assessed by the urinary function domain of the Prostate Cancer Index, and obstructive symptoms were assessed by the American Urological Association Symptom Index. Regression models were constructed to evaluate the impact of obstructive and incontinence symptoms on urinary impairment and satisfaction. RESULTS: The control group reported greater urinary continence (P <0.0001) and less impairment (P <0.0001) than the prostatectomy group, but a difference was not observed for obstructive symptoms. Obstructive urinary symptoms were associated with impairment for both the prostatectomy and the control groups (P <0.0001), and incontinence symptoms were more strongly associated with impairment in the prostatectomy group than in the control group (P = 0.01). Greater obstructive/irritative and incontinence symptoms were associated with lower satisfaction (P <0.0001). CONCLUSIONS: Urinary incontinence was more common in the radical prostatectomy group, but differences were not seen for obstructive symptoms. In addition to incontinence symptoms, obstructive symptoms were associated with urinary impairment and dissatisfaction in the prostatectomy group, suggesting that these symptoms should also be assessed after radical prostatectomy. PMID- 11880077 TI - Transrectal high-intensity focused ultrasound for treatment of patients with stage T1b-2n0m0 localized prostate cancer: a preliminary report. AB - OBJECTIVES: To present our preliminary clinical results of transrectal high intensity focused ultrasound (HIFU) in Stage T1b-2N0M0 prostate cancer. Efforts are being made to provide minimally invasive alternative treatment options with equal efficacy and fewer side effects. HIFU delivers ultrasound energy with rapid thermal necrosis of tissue in the focal region without damaging the surrounding tissue. METHODS: We performed 28 HIFU treatments in 20 patients with biopsy proven localized prostate cancer using a modified Sonablate-200 HIFU device. All patient characteristics and the clinical outcome of 20 patients followed up more than 6 months (mean 13.5) were analyzed. RESULTS: A complete response was obtained in 100% (20 of 20) of patients, as evidenced by a negative postoperative prostate biopsy and no elevation on three successive prostate-specific antigen (PSA) determinations. Of the 20 patients, 13 (65%), 5 (25%), and 2 (10%) had PSA nadirs of less than 0.50 ng/mL, 0.50 to 1.00 ng/mL, and 1.01 to 2.00 ng/mL, respectively. Rectourethral fistula and urethral stricture were noted in 1 and 2 patients, respectively, and 1 patient underwent transurethral resection of the prostate because of prolonged urinary retention. CONCLUSIONS: Our results show that HIFU can be performed without an incision, with a less severe side effect profile, and, unlike most other prostate treatments, is repeatable. Transrectal HIFU may be a useful option for patients with localized prostate cancer. Its long term efficacy will be determined by additional follow-up and a Phase II trial. PMID- 11880080 TI - Variations in use of imaging in a national sample of men with early-stage prostate cancer. AB - OBJECTIVES: To measure the national practice variations in imaging studies performed for men newly diagnosed with clinically localized prostate cancer. METHODS: We created an analytic file from 1991 to 1996 Medicare claims data using files for a random sample of 5% of all Medicare beneficiaries. Among men with newly diagnosed clinically localized prostate cancer, we identified those undergoing staging bone scans, staging computed tomography (CT), or staging magnetic resonance imaging (MRI) at the time of diagnosis. We conducted univariate and multivariate analyses adjusting for Charlson index score, age group, race, geographic region, and year of diagnosis. RESULTS: In all geographic regions, men receiving radiation therapy (RT) were more likely than those receiving radical prostatectomy (RP) to undergo CT. In the South, RT patients were significantly more likely than RP patients to undergo MRI and bone scans. In the West, RT patients were significantly more likely than RP patients to have bone scans. In multivariate analyses that controlled for all significant univariate findings, treatment with RT significantly predicted for the use of bone scans (odds ratio 1.24, 95% confidence interval 1.17 to 1.31), CT scans (odds ratio 3.26, 95% confidence interval 3.18 to 3.34), and MRI scans (odds ratio 1.47, 95% confidence interval 1.23 to 1.72). Regional differences in the use of imaging technologies for staging persisted in the multivariate analysis. CONCLUSIONS: Patients undergoing RT for clinically localized prostate cancer undergo more bone, CT, and MRI scans than do patients undergoing RP, regardless of comorbidity, age, or race. In addition, a significant geographic variation was found in the use of these diagnostic tests. These variations suggest that evidence-based staging guidelines have not been met with broad physician acceptance. PMID- 11880081 TI - Randomized prospective study of periprostatic local anesthesia during transrectal ultrasound-guided prostate biopsy. AB - OBJECTIVES: To prospectively evaluate the efficacy of apical and lateral periprostatic lidocaine injection as anesthesia during transrectal ultrasound guided prostate biopsy. METHODS: A total of 152 consecutive men undergoing transrectal prostate biopsy in our department were enrolled in this study. Patients were randomized into group 1 (74 patients) who received 15 mL of 1% lidocaine in the lateral and apical periprostatic regions, 5 mL in each point, 10 minutes before the prostate biopsy, and group 2 (78 patients), controls. Pain after each biopsy (a total of 10 biopsies) was assessed using a 10-point linear visual analog pain scale. RESULTS: In groups 1 and 2, the mean patient age was 64.8 and 65.8 years (P = 0.4), mean prostate-specific antigen was 10.9 and 11.2 ng/mL (P = 0.9), and mean prostate volume was 57 and 60 cm(3) (P = 0.5), respectively. The mean total pain score (sum of each biopsy score, total of 10 for each patient) during transrectal prostate biopsy was 16 (range 0 to 62.5) and 50 (range 0 to 100) in groups 1 and 2, respectively (P = 0.0001). No adverse events were noted. CONCLUSIONS: Transrectal ultrasound-guided lateral and apical periprostatic anesthesia significantly diminishes the pain in men undergoing transrectal prostate biopsy. We recommend that this procedure be routinely offered to all patients undergoing transrectal prostate biopsy. PMID- 11880082 TI - Prognostic value of p53 overexpression in T1G3 bladder tumors treated with bacillus Calmette-Guerin therapy. AB - OBJECTIVES: To evaluate the correlation between the overexpression of mutant protein p53 and disease recurrence and progression in patients treated with bacillus Calmette-Guerin (BCG) intravesical therapy for T1G3 bladder cancer. METHODS: We analyzed the outcome of 29 consecutive patients treated for T1G3 bladder tumor with transurethral resection. Patients previously treated for a bladder tumor, those who underwent incomplete resection, and those in whom no assessment of the muscle cell layer was possible were excluded from the study. p53 overexpression was determined using monoclonal p53-DO7 antibody, with a 20% cutoff for definition of positivity. After the initial transurethral resection, all patients were treated with Pasteur BCG (75 mg in 50 mL saline), weekly for 6 weeks. The correlation between p53 overexpression and disease recurrence and progression was assessed by the Fisher exact test. RESULTS: The median follow-up was 36.7 months (range 1 to 108). Of the 29 patients, 18 (62.1%) were p53 positive and 11 (37.9%) were p53 negative. Both groups were similar according to age, tumoral substage (T1a/T1b), association with carcinoma in situ, multifocality, and length of follow-up. The recurrence rate was 54.4% in the p53 negative group versus 38.9% in the p53-positive group (P = 0.47). The progression rate was 18.2% in the p53-negative group versus 33.3% in the p53-positive group (P = 0.67). CONCLUSIONS: These findings suggest that overexpression of p53, as determined immunohistochemically, has no predictive value for recurrence and progression in T1G3 bladder cancers treated with intravesical BCG. PMID- 11880084 TI - Magnetic resonance imaging of scrotal diseases: when it makes the difference. AB - OBJECTIVES: To investigate the utility of magnetic resonance imaging (MRI) after inconclusive sonography in the evaluation of scrotal disease. METHODS: From 1993 to 1999, 622 patients underwent ultrasonography (US) with state-of-the-art equipment to evaluate scrotal diseases. Twenty-six patients, two with bilateral and distinct diseases (age range 1--63 years, mean 33), had an inconclusive sonogram and were sent for MRI. The MRI results were compared with previous US reports and with the final diagnosis. RESULTS: MRI yielded additional and correct information (compared with US), coincident with the final diagnosis in 23 cases (82.1%). In regard to lesions suspected of malignancy at US (17 cases), MRI had a great concordance with the final diagnosis and was statistically significant (P <0.002, kappa test). CONCLUSIONS: The results of our study indicate that MRI could help elucidate scrotal dilemmas found at US, although the small percentage of inconclusive sonograms confirms this technique as the first choice when imaging is required in scrotal diseases. PMID- 11880083 TI - Long-term maintenance bacille Calmette-Guerin therapy in high-grade superficial bladder cancer. AB - OBJECTIVES: To assess the long-term results of intravesical bacille Calmette Guerin (BCG) induction plus long-term maintenance treatment for high-grade superficial bladder cancer. METHODS: Between 1994 and 2000, 41 patients who presented to our clinic with superficial urothelial carcinoma of the bladder (T1G3, T1G3 plus carcinoma in situ, or recurrent TaG2-3) were treated by transurethral resection of all visible tumor and an induction cycle of BCG plus a long-term maintenance BCG course consisting of 11 monthly instillations followed by 4 quarterly instillations and then by 6 six-monthly instillations. The median follow-up was 40 months. RESULTS: Thirty patients remained tumor free throughout the follow-up period. Ten patients had a recurrence of superficial tumor, 9 patients during the monthly instillation course and 1 patient during the quarterly instillation course. One patient presented with progression. CONCLUSIONS: Adjuvant immunotherapy with BCG after complete transurethral resection of bladder tumor represents a highly effective primary treatment for high-grade superficial bladder cancer. Our maintenance course of BCG seemed to improve the worldwide accepted effectiveness of the BCG induction course without any important side effects. PMID- 11880085 TI - Sterile water irrigation of the distal vas deferens at vasectomy: does it accelerate clearance of sperm? A prospective randomized trial. AB - OBJECTIVES: To assess the effect of irrigation of the distal vas deferens with sterile water at the time of vasectomy on sperm clearance. Is it possible to accelerate clearance and avoid the problem of lingering sperm? METHODS: Two hundred men undergoing vasectomy were randomized to receive either a standard vasectomy or vasectomy plus irrigation of each vas deferens with sterile water. The interval between vasectomy and the production of two azoospermic semen samples was recorded. RESULTS: Overall, 37 patients were excluded for failing to follow the postvasectomy protocol for semen analysis, leaving 87 in the control group and 76 who received vasal irrigation. No statistically significant differences existed between the two groups with regard to the mean time to clearance (26.4 weeks, control group versus 28.6 weeks, flush group), proportion clear at 16 weeks (29% versus 21%), or proportion with lingering sperm at 40 weeks (22% versus 26%). CONCLUSIONS: Irrigation of the vas deferens with sterile water does not accelerate the clearance of spermatozoa after vasectomy. Men wishing to undergo vasectomy must be counseled about the possibility of lingering sperm. PMID- 11880086 TI - Preliminary study of the safety and efficacy of extended-release oxybutynin in children. AB - OBJECTIVES: To test the safety and efficacy of extended-release oxybutynin in children with bladder dysfunction. The efficacy of oxybutynin in children has been limited by side effects. A new extended-release formulation of oxybutynin has some benefits versus traditional oxybutynin but has never been evaluated in children. METHODS: A retrospective study was performed on 25 children who had been treated with extended-release oxybutynin. Fourteen had neurogenic bladder dysfunction and 11 had urinary frequency and urgency and urge incontinence but no neurologic abnormalities. Patients and families were asked to semiquantitatively (0 to 10 grading with 10 = severe) assess the effects of the medication on efficacy, as well as side effects and compliance with medication schedules. RESULTS: All 25 patients had improvement in incontinence and/or voiding dysfunction on extended-release oxybutynin. Twelve (48%) experienced no side effects. Of the 13 who did, 10 complained of dry mouth (grade 4.6 plus minus 0.5), 4 had constipation (grade 5.8 plus minus 1.8), 4 had heat intolerance (grade 5.1 plus minus 0.9), and 3 had drowsiness (grade 5.3 plus minus 2.4). Of patients previously treated with oxybutynin, the extended-release oxybutynin was equally or more efficacious and had the same or fewer side effects, especially less dry mouth. Families reported much better patient compliance with the medication regimen using extended-release oxybutynin compared with oxybutynin. Patient and family satisfaction was very high, and 21 of 25 have continued using the medication. CONCLUSIONS: Extended-release oxybutynin is safe and efficacious in children. In this preliminary evaluation, it had benefits over traditional, immediate-release oxybutynin. PMID- 11880087 TI - Relationships among nocturnal urinary volume, bladder capacity, and nocturia with and without water load in nonenuretic children. AB - OBJECTIVES: To evaluate the relationships among nocturnal urinary volume (NUV), bladder capacity, and nocturia in nonenuretic children with and without water load. METHODS: We evaluated 72 healthy nonenuretic volunteers. The NUV and bladder capacity were measured for 2 nights at volunteers' homes with and without water load. RESULTS: Of the 72 nonenuretic children, 45 (63%) were classified into the large NUV group in which the NUV with water load was more than the upper limit of the 95% confidence interval of NUV in Japanese nonenuretic children, and the other 27 (37%) were classified into the small NUV group. In the large NUV group, 33 (73%) had one or two episodes of nocturia because of water intake, and 12 (27%) did not. The nocturnal bladder capacity in the latter 12 children was significantly larger than that in the former 33, although no significant differences were found in functional bladder capacity and NUV with water intake. CONCLUSIONS: Most nonenuretic children were able to cope with the physiologic loading of water that would cause bedwetting in enuretic children. After water loading, three quarters of the nonenuretic children were able to awake easily because of the need to urinate with a certain volume of urine in the bladder, and the one quarter who did not awake were able to hold more urine in their bladder in the sleep state. PMID- 11880089 TI - Patient positioning and port placement for bilateral hand-assisted laparoscopic nephrectomy. AB - Hand-assisted laparoscopic nephrectomy is an alternative to laparoscopic nephrectomy. We describe our method for triangular port placement and patient positioning during bilateral hand-assisted laparoscopic nephrectomy. This method does not require intraoperative patient repositioning and may decrease the overall operative time while enhancing patient safety. PMID- 11880088 TI - Congenital isolated penile torsion in adults: untwist with plication. AB - To correct congenital isolated penile torsion, we delineated a parabolic curve as a counterdirection to the torsion after artificial erection was induced. Two areas along the curve were chosen and plicated with two interrupted U-shaped sutures. Satisfactory functional and cosmetic results were achieved in 3 patients. PMID- 11880090 TI - Long-term recurrence of primary amyloidosis of the bladder. AB - An 84-year-old man presented to our department with a recurrence of a primary amyloidosis of the bladder 14 years after the first diagnosis. Follow-up had been stopped 5 years after the first procedure because no cystoscopic anomalies were noted. General evaluation confirmed the diagnosis of primary amyloidosis. The lesions were endoscopically resected. At the 3-year follow-up, the patient was free of symptoms and had remained cystoscopically free of disease. However, stable amyloidosis deposits have always been noted on the computed tomography scans. This case emphasizes the necessity of a longer follow-up, including computed tomography of the pelvis, during the 10 to 15 years after the first occurrence of the disease. PMID- 11880091 TI - Posterior approach for retroperitoneal laparoscopic bilateral nephrectomy in a child. AB - We report on a unique posterior approach for bilateral nephrectomy by retroperitoneoscopy that was conducted in a 13-year-old boy with end-stage renal disease managed by peritoneal dialysis. A posterior approach for bilateral nephrectomy by retroperitoneal laparoscopy provided adequate visualization and created enough working space for the manipulation. The procedure was performed during a short period, and the patient did not need to be repositioned. Retroperitoneal laparoscopic bilateral nephrectomy is a useful option in school aged children with poorly functioning kidneys, particularly those children also undergoing peritoneal dialysis. PMID- 11880092 TI - Pulmonary embolus related to PC-SPES use in a patient with PSA recurrence after radical prostatectomy. AB - Patients diagnosed with a rising prostate-specific antigen level after radical prostatectomy represent a therapeutic dilemma. Herbal therapies including PC-SPES have gained popularity as alternatives to conventional hormonal ablation in such patients as a result of the perceived benefits of using natural products and proven clinical response in decreasing prostate-specific antigen levels. PC-SPES is one of the most popular herbs with known estrogenic activity in vitro and in vivo. Estrogenic compounds increase the risk of thromboembolic events. However, no specific guidelines are available with regard to the risk of thromboembolic events for patients using PC-SPES. We report a case of a patient treated with PC SPES for prostate-specific antigen recurrence after radical prostatectomy who presented with pulmonary embolus and a right common femoral deep venous thrombus. PMID- 11880093 TI - Irritative voiding symptoms and microscopic hematuria caused by intraperitoneal calcified fat necrosis. AB - A previously healthy 60-year-old man presented with urinary urgency and microhematuria. The workup revealed no urothelial lesions, but did suggest a calcified intraperitoneal mass causing extrinsic compression of the bladder. Laparoscopic exploration revealed a glistening, spherical mass attached by a stalk to the sigmoid colon. Removal and histologic examination of the mass revealed calcified fat necrosis, most likely due to the spontaneous torsion and calcification of an appendix epiploicae. The patient's symptoms and microhematuria resolved after removal. This case represents an unusual occurrence of irritative voiding symptoms probably due to the extrinsic compression of the bladder by a mass of membranous fat necrosis. PMID- 11880094 TI - Laparoscopic excision of a solitary renal cell carcinoma metastasis to the contralateral perirenal adipose tissue. AB - Metastasis of renal cell carcinoma to the contralateral perirenal fat is a very rare occurrence. We report a case of a synchronous, solitary perirenal metastasis excised laparoscopically 2 years after initial, open radical nephrectomy. PMID- 11880095 TI - Successful angioembolization of renal artery pseudoaneurysms after blunt abdominal trauma. AB - We report a case of minimally invasive management of renal artery pseudoaneurysms after blunt abdominal trauma. A 44-year-old man developed gross hematuria after a motor vehicle collision. The initial radiographic evaluation revealed only a parenchymal laceration of the right kidney, and the patient was treated conservatively. Persistent gross hematuria necessitated repeat renal angiography that revealed numerous small pseudoaneurysms. Angioembolization was successfully performed with minimal compromise of the renal parenchyma. Transcatheter embolization is safe and effective for controlling hemorrhage from traumatic renal artery pseudoaneurysms and should be the initial treatment of choice in hemodynamically stable patients. PMID- 11880096 TI - Primary squamous cell carcinoma of the seminal vesicle. AB - A 69-year-old man with primary squamous cell carcinoma of the seminal vesicle was successfully treated. The patient presented with complaints of gross hematuria and difficulty in urination. Transrectal ultrasonography revealed a cystic mass behind the bladder, and solid tumorous lesions were visualized in the cyst wall by magnetic resonance imaging. The cystic mass was surgically excised and was verified as the enlarged seminal vesicle with concomitant squamous cell carcinoma. The histopathologic diagnosis was squamous cell carcinoma in the seminal vesicle. Postoperatively, 50 Gy of external beam radiation was targeted to the whole pelvic cavity, and the patient was alive without recurrence 2.5 years after surgery. No other neoplasm was discovered in the body. PMID- 11880097 TI - Leiomyoma of the corona glans penis. AB - A 42-year-old man presented with an 8-mm elliptical indolent solid mass on the superior surface of the corona glandis at our institution 3 months after its appearance. The lesion was excised by using a wedge resection performed by a cool blade knife. The subsequent pathologic evaluation defined the lesion as a leiomyoma. Immunohistochemical analysis using the streptavidin-biotin peroxidase technique and immunoreactivity for cytokeratin, S100 protein, and the proliferative marker MIB1 was performed, confirming the pathologic diagnosis. Early local excision of these tumors is indicated because of the trend toward their dimensions increasing quickly, with subsequent difficulties for cosmetic glandis reconstruction. PMID- 11880098 TI - Primary testicular amyloidosis mimicking tumor in a cryptorchid testis. AB - We present a case of primary testicular amyloidosis mimicking tumor in a cryptorchid testis. Examination revealed the absence of a palpable testis in the left hemiscrotum. The laboratory evaluation showed azoospermia and a low testosterone level. The radiologic investigation demonstrated a heterogeneous left inguinal mass suspicious for tumor in a cryptorchid testis. After radical orchiectomy, the histologic evaluation revealed primary amyloidosis of the testis. The pathogenesis and clinical presentation of testicular amyloidosis are reviewed. PMID- 11880099 TI - An unusual case of penile augmentation: subcutaneous stone implantation. AB - Different techniques for penile augmentation have been published. We report a 45 year-old man who had undergone a subcutaneous penile stone implantation for enlargement, which is the first published report to our knowledge. PMID- 11880100 TI - Transvaginal laparoscopic nephrectomy: development and feasibility in the porcine model. AB - OBJECTIVES: To assess feasibility of laparoscopic nephrectomy completed entirely by way of the vagina in the porcine model. METHODS: Six transvaginal laparoscopic nephrectomies were performed in female farm pigs. Two acute and two 1-week survival animals were used for the study. Before killing the survival animals, a second transvaginal laparoscopic nephrectomy was performed on the remaining renal unit. For one renal unit, the laparoscopic nephrectomy was completed entirely by way of the vagina. In five renal units, a single, 5-mm transabdominal trocar for the laparoscope was required to facilitate visualization. RESULTS: The operative time for the procedure completed entirely by way of the vagina was 360 minutes, and the mean operative time for the procedures requiring placement of a single 5 mm transabdominal trocar was 210 minutes. In 5 cases, dissection, control of the renal pedicle, and extraction of the kidney were successfully completed using a transvaginal approach. In 1 acute case, an uncontrollable vascular injury occurred during placement of the Endo-GIA stapler, resulting in exsanguination. In all other cases, the mean blood loss was less than 30 mL, and no significant perioperative complications were noted. Both survival pigs had normal bowel and bladder function before being killed. CONCLUSIONS: Complete transvaginal laparoscopic dissection and nephrectomy is feasible in the porcine model using a single, 5-mm abdominal trocar for visualization. A completely transvaginal laparoscopic nephrectomy was performed once, but limitations imposed by the porcine anatomy and by the currently available instrumentation made the procedure very cumbersome. Additional development of this technique in animal models and improved instrumentation is needed before clinical assessment is warranted. PMID- 11880101 TI - Gene expression profiles in the Peyronie's disease plaque. AB - OBJECTIVES: To provide molecular insight into the pathophysiology of Peyronie's disease (PD), a preliminary profile of differential gene expression between the PD plaque and control tunica albuginea was obtained with DNA microarrays. METHODS: Seven PD plaques and five control tunica albugineas were studied. cDNA specimens were prepared from RNA isolated from one calcified PD plaque and one control tissue and hybridized with the Clontech Atlas 1.2 Array. Another set of plaque and control RNA samples was hybridized with the Affymetrix GeneChip. Relative changes of greater than 2.0 defined up-regulation and down-regulation, respectively. RNA from the remaining tissues was used to determine, by reverse transcriptase-polymerase chain reaction and Western blot analysis, the expression of selected individual genes. RESULTS: Some of up-regulated genes in the PD plaque detected by the Clontech assay were pleiotrophin, monocyte chemotactic protein 1, and early growth response protein, which are involved in osteoblast recruitment, inflammation, and fibroblast proliferation, respectively. Ubiquitin and Id-2, which are involved in tissue remodeling, were down-regulated. The Affymetrix DNA chips identified the up-regulation of elastase (involved in elastic fiber degradation) and the myofibroblast markers alpha and gamma-smooth muscle actin, desmin, and others, as well as the down-regulation of collagenase IV and transforming growth factor-beta modulators. Four of the five genes selected for reverse transcriptase-polymerase chain reaction and Western blotting confirmed the DNA microarray results. CONCLUSIONS: In the PD tissue, the genes involved in collagen synthesis, myofibroblast differentiation, tissue remodeling, inflammation, ossification, and proteolysis are up-regulated, and the genes that inhibit some of these processes and collagenase are down-regulated. PMID- 11880102 TI - Urokinase gene 3'-UTR T/C polymorphism is associated with urolithiasis. AB - OBJECTIVES: To study single nucleotide polymorphisms to investigate the possibility that urokinase is involved in the formation of urolithiasis, which, although lacking in genetic evidence, has been previously proposed. METHODS: A total of 153 patients with recurrent calcium stones and 105 controls were studied. Polymerase chain reaction-based restriction analysis was used to identify the C/T polymorphism of the urokinase gene, which is mapped on the 3' untranslated region (3'-UTR) on chromosome 10. RESULTS: A significant difference was found in the distribution of the urokinase gene 3'-UTR C/T polymorphism frequency between patients with stones and normal controls (P <0.05). The odds ratio for the risk of the "T" allele in patients with stones was 3.088 (95% confidence interval 1.06 to 8.99). CONCLUSIONS: The results of our study demonstrate that the urokinase gene 3'-UTR "T" allele is associated with calcium stone disease. Individuals possessing the "T" allele have a higher incidence of calcium oxalate stone disease. The results of this study provide genetic evidence that the urokinase gene may play a role in stone formation. PMID- 11880104 TI - "Bazex syndromes": dissimilar entities named after the same father! PMID- 11880103 TI - Low levels of PSA predict long-term risk of prostate cancer: results from the Baltimore longitudinal study of aging. PMID- 11880107 TI - Detection and diagnosis of blood in feces and urine: an overview. AB - BACKGROUND: Detection of blood in feces or urine has long been regarded as an indicator of patient's state of health. The ease with which feces or urine may be obtained and patient's willingness to provide the specimen make detection of fecal occult blood or urine analysis one of the most commonly performed screening examinations. Historically, the inspection of feces or urine for diagnostic purpose has been practiced for centuries. Of late, management of renal or urinary tract abnormalities or investigation of anemia, gastrointestinal diseases and for early detection of colorectal cancer has assumed greater importance. METHODS: The never-ending list of techniques for the diagnosis of disorders producing bleeding such as urine microscopy, urine cytology, urine based marker test, cytoscopy, ultra sonography, computed tomography, magnetic resonance imaging, to mention a few, and four categories of detection of fecal occult blood namely, radioanalytical, physical, immunochemical and chemical methods makes the study very interesting. PURPOSE: This review attempts to overview various techniques, methods and methodologies for the diagnosis and detection of blood in feces and urine, in the direction of looking at past and current tests with an eye on future needs. PMID- 11880108 TI - Interference-free sample preparation for the determination of plasma oxalate analyzed by HPLC-ER: preliminary results from calcium oxalate stone-formers and non-stone-formers. AB - BACKGROUND: Oxalate generation at pH-values above 5.0 and an oxalate-protein binding in acidified plasma would appear to complicate the determination of oxalate in plasma. METHODS: To avoid complex sample preparation we used a high performance liquid chromatographic system with an inline enzyme reactor (HPLC-ER) containing immobilised oxalate oxidase. The detection limit was 0.68 micromol/l. Blood was drawn in lithium-heparin vessels and immediately centrifuged at 4 degrees C. The yielded plasma was ultrafiltered using a Centrisart-I-tube. To inhibit oxalate generation by ascorbic acid, the ultrafiltrate was acidified with 1 mol/l hydrochloric acid during ultrafiltration at 4 degrees C. The liquid thus yielded was used for HPLC-ER analysis. Blood samples were obtained from 133 healthy adults (63 men, 70 women, aged 20-94 years) with no history of renal disorder and from 79 patients (53 men, 26 women, aged 19-77 years) with a history of calcium oxalate stone formation. RESULTS: Mean plasma oxalate was 2.65 +/- 2.31 micromol/l for healthy subjects and 4.21 +/- 0.56 micromol/l for stone formers. CONCLUSIONS: Analysis yielded no significant differences between males and females. A correlation between age and plasma oxalate was found for the healthy adults (p < 0.001). PMID- 11880109 TI - A novel monoclonal antibody to fibrin monomer and soluble fibrin for the detection of soluble fibrin in plasma. AB - BACKGROUND: Soluble fibrin (SF), composed of fibrin monomer (FM) and fibrinogen, is well known to exist in the circulating blood derived from patients with thrombotic diseases, and its quantification is useful to get some information on the state and degree of intravascular coagulation. However, there was no convenient method for the determination of SF. METHODS: We prepared a novel monoclonal antibody (MoAb) (F405) to FM and SF using desAA-fibrin as the immunogen in the presence of anti-polymerant peptide (Gly-Pro-Arg-Pro, GPRP), and the characterization of the F405 was performed by Western blotting analysis and an enzyme-linked immunosorbent assay (ELISA). We also tried to detect SF in human plasma using an ELISA involving the immobilized F405 and horseradish peroxidase (POD)-labeled anti-fibrinogen polyclonal antibody. RESULTS: The antibody reacted with the fibrin degradation products fragments X, Y and E, but not with fibrinogen or its fragments X, Y, D and E, or the fibrin D-dimer. The epitope recognized by F405 appeared to be the alpha-chain N-terminal region exposed upon removal of the A peptide from the Aalpha-chain because F405 was found to bind to the alpha-chain N-terminal oligo-peptide of fibrin (GPRVVERHQ). Since F405 reacted not only with FM in the presence of GPRP peptide, but also with the SF complex prepared by the addition of thrombin-treated FM to human fibrinogen, we attempted to detect SF in human plasma using ELISA. The analytical range of this method was 1-300 microg/ml. The assay detection limit was < 0.5 microg/ml, and the results of intra- and inter-assay precision studies indicated that this method is accurate and yields reproducible results (< 9.4% and < 10%, respectively). When 56 samples of plasma from patients with disseminated intravascular coagulation (DIC) and 117 control samples from healthy individuals were tested, elevated levels of SF complex were detected in the DIC samples: the mean +/- S.D. of the SF concentration in the DIC and control samples were 63.4 +/ 65.3 microg/ml and 1.9 +/- 1.0 microg/ml, respectively. CONCLUSIONS: The ELISA using F405 is useful for the diagnosis of DIC. PMID- 11880110 TI - Real-time quantitative reverse transcription-PCR assay for renal cell carcinoma associated antigen G250. AB - OBJECTIVES: Gene amplification/expression of G250 is a major event in human renal tumorigenesis. G250-based therapeutic agents and G250-specific gene therapy are under development. These new perspectives call for a sensitive and accurate method to screen G250 alterations in renal cell cancer (RCC) patients and investigate the relationship between G250 mRNA expression and RCC. METHODS: We developed a quantitative RT-PCR assay for the measurement of G250 mRNA expression using a real-time procedure based on the use of fluorogenic probes and the ABI PRISM 7700 Sequence Detector System. The method has been applied to the measurement of quantitative mRNA level of G250 in 31 cases RCC and 6 normal renal tissues. RESULTS: The dynamic range was 10(3)-10(8). The relationship between Ct and log starting concentration was linear (r=0.99). G250 expression was present in all RCCs with G250 amplification but was absent in normal ones. G250 mRNA expression ranged from 2.9 x 10(3) to 6.5 x 10(7) copy/microg RNA, with a mean value of 3.5 x 10(6) copy/microg RNA. The expression of G250 revealed an inverse correlation to tumor grade. G250 mRNA level did not correlate with the cell types and clinical stages (P>0.05). CONCLUSIONS: G250 has the potential to be used as a marker of diagnosis and increasing proliferation in RCC. This new simple, rapid, semi-automated assay was a major alternative to competitive PCR and Northern blot analysis for gene alteration analysis in human tumors and might be a powerful tool for large randomized, prospective cooperative group trials and supporting future G250-based biological and gene therapy approaches. PMID- 11880111 TI - Detection of human papillomavirus DNA by PCR/microfluorometry for screening of cervical cancer. AB - BACKGROUND: Cervical cancer screening is conducted by a cytological Papanicolaou (Pap) test. For screening, it is becoming increasingly important to introduce a more objective result, based on human papillomavirus (HPV) DNA test. We describe here a practical method allowing the mass detection of HPV-DNA by PCR followed by fluorogenic DNA intercalation. METHODS: Samples used were cervical scrapes or biopsy specimens obtained from women who had undergone cytological testing for cervical cancer. Crude DNAs were extracted by a simplified proteinase K-boil method. Common and type-specific primers were newly designed for major types of high-risk HPVs. A fluorogenic DNA intercalator, SYBR Green I was directly added to the specific PCR products. The resultant fluorescence was measured by a conventional fluorometric microplate reader. RESULTS: The proposed PCR/microfluorometry (MFL) allowed a simple, rapid and economical detection of HPV-DNA without any use of labeling primers or probes. HPV-DNAs were found in 48.2% (123/255) of the cervical scrapes. The detection rate of HPV in cervical cancer biopsy specimen was 92.4% (61/66). CONCLUSIONS: PCR/MFL detection of HPV DNA, followed by combined type-specific PCR, is expected to be an extremely useful tool in cervical cancer screening. PMID- 11880112 TI - Long-term administration of L-carnitine to humans: effect on skeletal muscle carnitine content and physical performance. AB - BACKGROUND: Long-term administration of high oral doses of L-carnitine on the skeletal muscle composition and the physical performance has not been studied in humans. METHODS: Eight healthy male adults were treated with 2 x 2 g of L carnitine per day for 3 months. Muscle biopsies and exercise tests were performed before, immediately after, and 2 months after the treatment. Exercise tests were performed using a bicycle ergometer for 10 min at 20%, 40%, and 60% of the individual maximal workload (P(max)), respectively, until exhaustion. RESULTS: There were no significant differences between V(O(2)max), RER(max), and P(max) between the three time points investigated. At submaximal intensities, the only difference to the pretreatment values was a 5% increase in V(O(2)) at 20% and 40% of P(max) 2 months after the cessation of the treatment. The total carnitine content in the skeletal muscle was 4.10 +/- 0.82 micromol/g before, 4.79 +/- 1.19 micromol/g immediately after, and 4.19 +/- 0.61 micromol/g wet weight 2 months after the treatment (no significant difference). Activities of the two mitochondrial enzymes citrate synthase and cytochrome oxidase, as well as the skeletal muscle fiber composition also remained unaffected by the administration of L-carnitine. CONCLUSIONS: Long-term oral treatment of healthy adults with L carnitine is not associated with a significant increase in the muscle carnitine content, mitochondrial proliferation, or physical performance. Beneficial effects of the long-term treatment with L-carnitine on the physical performance of healthy adults cannot be explained by an increase in the carnitine muscle stores. PMID- 11880113 TI - Assay of serum allantoin in humans by gas chromatography-mass spectrometry. AB - BACKGROUND: The small amount of allantoin present in human serum results from free radical (FR) action on urate and may provide a stable marker of free radical activity in vivo. We describe a gas chromatography-mass spectrometry (GC-MS) assay for serum allantoin and report a reference range in healthy individuals. METHODS: Fasting blood samples were obtained from 134 healthy middle-aged volunteers (56 men, mean age 55, range 45-72; 78 women, mean age 55, range 50-72) Allantoin was assayed using 15N(2) allantoin as an internal standard. After isolation from aqueous standards or serum by extraction onto an anion exchange column (AG-MP1), allantoin was derivatised with N-methyl-N-(tert butyldimethylsilyl) trifluoroacetamide (MTBSTFA). Derivatives were injected onto an HP-1 column and analysed using a Mass Selective Detector with Single Ion Monitoring at 398 and 400 m/z. RESULTS: The distribution of serum allantoin concentrations in men and women was non-Gaussian and log transformation was used for the analysis of data. Women (10.8 +/- 1.7 micromol/l (mean +/- S.D.)) had significantly lower serum allantoin levels than men (13.4 +/- 1.6 micromol/l, p=0.015). Reference ranges (95% CI) for middle-aged healthy subjects were 7.4 46.8 micromol/l (men) and 3.7-31.2 micromol/l (women). CONCLUSION: Gas chromatography-mass spectrometry provides a reliable and accurate method for the determination of serum allantoin. PMID- 11880114 TI - Influence of mycophenolic acid on inosine 5'-monophosphate dehydrogenase activity in human peripheral blood mononuclear cells. AB - BACKGROUND: Inosine 5'-monophosphate dehydrogenase (IMPDH) catalyses the oxidation of inosine 5'-monophosphate (IMP) to xanthosine 5'-monophosphate (XMP). Thus, this enzyme plays an important role in the rate-limiting step of the de novo guanine nucleotide biosynthesis, making it a potent target for immunosuppressive drugs. Mycophenolic acid (MPA) is the most potent and specific inhibitor of IMPDH. METHOD: IMPDH activity is determined via evaluation of XMP formation and the inhibitory influence of MPA in human peripheral blood mononuclear cells (PBMCs) is assessed by means of high-performance liquid chromatography (HPLC). For this objective, we have optimised a method based on solvent-generated ion exchange chromatography by cautiously varying mobile phase parameters. RESULTS: The optimised method renders it possible to separate 18 analytes in 54 min in a single isocratic experiment and to measure the IMPDH activity in the lysate of human PBMCs in dependence on incubation time, substrate, co-substrate and inhibitor concentrations. In this way, we have determined the Michaelis-Menten constants K(M) and V(max) for IMP and beta-NAD+ and the inhibitor constant K(i) for MPA. CONCLUSIONS: The chromatographic method presented in this report allows a rapid, reliable and reproducible quantification of IMPDH activity in PBMCs and therefore represents an attractive tool for the pharmacodynamic monitoring of the effects of MPA in patients under immunosuppressive therapy. PMID- 11880115 TI - Nitric oxide in ischemic and reperfused human muscle. AB - BACKGROUND: Biochemical events explaining the pathology of ischemia-reperfusion in the muscle are still debated. Nitric oxide (NO) has been postulated to be implicated in these phenomena, but the short half-life of this compound makes it difficult to measure. METHODS: In this paper, we used an amperometric solid-sate sensor to measure NO concentrations in frozen human muscles before, during and after a period of ischemia. We also measured cytochrome oxidase activity and malondialdehyde (MDA). RESULTS: NO increased during ischemia but it soon returned to normal values upon reperfusion. On the other hand, cytochrome oxidase that also decreased in ischemic muscle did not increase during the reperfusion and malondialdehyde only increased during reperfusion, indicating the occurrence of peroxidative reactions in this situation. CONCLUSIONS: NO is implicated in the ischemia/reperfusion pathology, but it is difficult to relate whether this is connected to cytochrome oxidase activity and malondialdehyde formation, also modified in this ischemia-reperfusion model. PMID- 11880116 TI - Effect of CuCl2, NaCl and EDTA on the enzyme alpha-L-iduronidase in the plasma of normal individuals and heterozygotes for MPS I. AB - BACKGROUND: It has been previously demonstrated that the enzyme alpha-L iduronidase (IDUA) of patients with MPS I shows a different biochemical behavior in each of the three clinical forms of these. In heterozygotes, its biochemical behavior has been recently established in leukocyte and plasma samples, demonstrating that it is possible to distinguish individuals heterozygous for MPS I within an unselected population. METHODS: We evaluated the effect of copper chloride, EDTA and sodium chloride on the activity of the enzyme alpha-L iduronidase in the plasma of normal individuals and of MPS I heterozygotes and observed the type of inhibition caused, the Ki, the apparent Km and the apparent Vmax for each inhibitor. RESULTS: Sodium chloride inhibited the enzyme in normal individuals and in 40% of the heterozygotes evaluated and activated it in 60% of heterozygotes. The remaining compounds inhibited IDUA in both heterozygotes and normal individuals. CONCLUSIONS: We detected significant differences capable of differentiating MPS I heterozygotes from normal individuals by simply adding sodium chloride, EDTA or copper chloride to the incubation medium at the time of IDUA activity determination, with a potential use in carrier detection protocols. PMID- 11880117 TI - Marked removal of bezafibrate-induced high-density lipoprotein-cholesterol by low density lipoprotein apheresis. AB - BACKGROUND: A case of marked reduction of the bezafibrate-induced increase of high-density lipoprotein (HDL)-cholesterol by low-density lipoprotein apheresis (LDL-apheresis) has not been previously reported. METHODS: A 68-year-old Japanese man with arteriosclerosis obliterans (ASO), diabetes mellitus, and hyperlipidemia underwent LDL-apheresis, followed by the concomitant bezafibrate administration. Plasma lipids of pre- and post-LDL-apheresis were measured and apolipoprotein E (apoE) localization of the pre- and post-LDL-apheresis was detected by agarose gel electrophoresis. RESULTS: Plasma concentrations of the total cholesterol, LDL cholesterol, triglyceride, and HDL-cholesterol of pre-LDL-apheresis were 4.78 +/- 0.36, 2.74 +/- 0.24, 2.44 +/- 0.52, and 0.92 +/- 0.10 mmol/l, respectively; those of the post-LDL-apheresis were 1.94 +/- 0.31, 0.72 +/- 0.13, 0.81 +/- 0.38, and 0.86 +/- 0.11 mmol/l, respectively. LDL-apheresis reduced HDL-cholesterol by 6.4% (p=0.346). During the bezafibrate administration, plasma concentrations of the above of pre-LDL-apheresis were 5.24 +/- 0.34, 3.28 +/- 0.22, 1.26 +/- 0.25, and 1.39 +/- 0.21 mmol/l, respectively; those of the post-LDL-apheresis were 2.25 +/- 0.44, 0.80 +/- 0.12, 0.58 +/- 0.19, and 1.18 +/- 0.16 mmol/l, respectively. LDL apheresis reduced HDL-cholesterol by 15.2% (p<0.01). Plasma apolipoprotein E detected between the prebeta- and alpha-mobility was markedly lower after the LDL apheresis in the agarose gel electrophoresis. CONCLUSIONS: The removal of the bezafibrate induced an increase of the HDL-cholesterol by LDL-apheresis. PMID- 11880118 TI - Detection of DNA mutations associated with mitochondrial diseases by Agilent 2100 bioanalyzer. AB - BACKGROUND: Molecular analysis of mitochondrial DNA (mtDNA) has provided a final diagnosis for many of the mitochondrial diseases. We evaluated the Agilent 2100 bioanalyzer (Agilent Technologies, Palo Alto, CA) to determine whether the system could replace the conventional restriction fragment length polymorphism (RFLP) analysis by the agarose gel electrophoresis for the detection of the mtDNA mutation. METHODS: Three members of a family with MELAS syndrome and four members of a family with MERRF syndrome were recruited for this study. After PCR and restriction enzyme digestion, DNA fragments were separated on the Agilent 2100 bioanalyzer in conjunction with the DNA 500 and DNA 1000 Labchip kits and by electrophoresis on precast 3% agarose gels. RESULTS: The data generated by the DNA 500 and DNA 1000 assays using the Agilent 2100 bioanalyzer showed a lower percentage error and a better reproducibility as compared to those obtained by the conventional method. CONCLUSION: Based on the performance of the bioanalyzer, we suggest that this novel Labchip is adequate to replace the current RFLP analysis by the agarose gel electrophoresis for mtDNA mutation detection. PMID- 11880119 TI - A simple method of detecting K-ras point mutations in stool samples for colorectal cancer screening using one-step polymerase chain reaction/restriction fragment length polymorphism analysis. AB - BACKGROUND: We examined a technique for detecting point mutations of K-ras codon 12 in stool samples using one-step polymerase chain reaction/restriction fragment length polymorphism (PCR/RFLP) analysis, in order to determine whether it could be used to screen for colorectal cancer. METHODS: DNA was extracted from 200-mg stool specimens of 5 healthy controls and 31 colorectal cancer patients. A 107 base-pair fragment of exon 1 of K-ras was amplified by PCR using mismatched primers. PCR products were digested with Bst NI and analyzed by gel electrophoresis followed by silver staining. Specificity of one-step PCR/RFLP was examined by using synthetic oligonucleotides. The detection limit of K-ras codon 12 mutations was determined by using SW480 and HT29 cells. RESULTS: The K-ras gene was successfully amplified from all healthy controls and colorectal cancer patients studied. Mutations of K-ras codon 12 were not detected in any of the healthy controls, but were identified in 13 (41.9%) of the 31 patients with colorectal cancer. Mutations were detectable in all six synthetic mutant DNAs, while none were detected among the wild type. The detection limit of this method was > or = 0.1%. CONCLUSIONS: PCR/RFLP analysis could be used in mass screening for colorectal cancer, because it is highly specific, has a low detection limit, and is simpler than conventional methods for detecting genetic abnormalities. PMID- 11880121 TI - Significantly decreased extracellular magnesium in brains of gerbils subjected to cerebral ischemia. AB - BACKGROUND: A method for determining extracellular magnesium (Mg) levels in gerbil brain dialysates was developed by microdialysis and graphite furnace atomic absorption spectroscopy (GFAAS). METHODS: Two microdialysis probes were inserted into the right and left cortices of a gerbil subjected to a focal cerebral ischemia. Extracellular magnesium concentrations in diluted dialysates were 1.10 and 1.12 ppb in the ipsilateral and contralateral gerbil cortex, respectively. RESULTS: During cerebral ischemia, these concentrations decreased significantly to approximately 60% of basal in the ipsilateral cortex, whereas no changes in the contralateral cortex were detected. Extracellular magnesium concentrations returned to baseline within 3 h of reperfusion. The linearity of magnesium concentrations ranged from 0.50 to 5.0 ppb with a detection limit of 0.03 ppb in the present assay. A complete analysis can be performed within 2 min. The intra- and interassay precision was < 5%. CONCLUSION: To our knowledge, the present method is the first analytical assay measuring dynamic extracellular magnesium concentrations during cerebral ischemia by microdialysis and graphite furnace atomic absorption spectroscopy. PMID- 11880120 TI - Activation of inducible nitric oxide synthase by Euonymus alatus in mouse peritoneal macrophages. AB - BACKGROUND: Euonymus alatus (EA) has been used for tumor therapy. However, it is still unclear how this herb prevents the diseases in experimental models. Nitric oxide (NO) as a potent macrophage-derived effector molecule against a variety of tumors has received increasing attention. METHODS: Using mouse peritoneal macrophages, we have examined the mechanism by which EA regulates NO production. RESULTS: When EA was used in combination with recombinant interferon-gamma (rIFN gamma), there was a marked cooperative induction of NO production. However, EA had no effect on NO production by itself. The increased production of NO from rIFN-gamma plus EA-stimulated cells was almost completely inhibited by pre treatment with pyrrolidine dithiocarbamate (PDTC), an inhibitor of nuclear factor kappa B (NF-kappaB). Furthermore, treatment of peritoneal macrophages with rIFN gamma plus EA caused a significant increase in tumor necrosis factor-alpha (TNF alpha) production. PDTC also decreased the effects of EA on TNF-alpha production significantly. CONCLUSIONS: EA increases the production of NO and TNF-alpha by rIFN-gamma-primed macrophages and suggest that NF-kappaB plays a critical role in mediating these effects of EA. PMID- 11880122 TI - Anticoagulant effects on plasma coenzyme Q(10) estimated by HPLC with coulometric detection. AB - BACKGROUND: The proportion of reduced coenzyme Q(10) (Q(10)H(2)) in total coenzyme Q(10) (TQ(10)), referred to as the Q(10)H(2):TQ(10) ratio, may be used as a possible marker of in vivo oxidative stress. However, the ranges for Q(10)H(2):TQ(10) ratio from previous reports are quite variable. Sample handling and preparation appear to have a profound effect on the stability of Q(10)H(2). METHODS: Paired tests were used to estimate TQ(10), Q(10)H(2), oxidized coenzyme Q(10) (Q(10)), and Q(10)H(2):TQ(10) ratio in patient samples collected in vacutainers containing heparin or EDTA. Sample tubes were immediately placed on ice and promptly centrifuged. After harvesting plasma, 100 microl of plasma was extracted with 1-propanol and centrifuged. The supernatant was injected directly into a high-performance liquid chromatography (HPLC) system. RESULTS: Significantly higher values (p=0.0015) of TQ(10), Q(10)H(2), and Q(10)H(2):TQ(10) ratio were noted in heparinized plasma as compared to EDTA plasma; Q(10) concentrations were lower in heparinized plasma. When vacutainers containing specimen were opened and kept refrigerated, the Q(10)H(2):TQ(10) ratios in heparinized samples were stable over 7 h with variation <3%. Blood Q(10)H(2) in closed heparin vacutainers kept refrigerated was stable up to 24 h. CONCLUSIONS: Our results indicate that the heparinized plasma is superior to the EDTA plasma in all measurements for coenzyme Q(10). PMID- 11880123 TI - Tay-Sachs and Sandhoff diseases: enzymatic diagnosis in dried blood spots on filter paper: retrospective diagnoses in newborn-screening cards. AB - BACKGROUND: Tay-Sachs disease (TSD), Sandhoff disease (SD) and variants are caused by deficient activity of the lysosomal enzymes hexosaminidase A (HA) and total hexosaminidase (TH) (hexosaminidase A plus B), respectively. For diagnosis, these enzymes are usually measured in plasma or extracts of leukocytes. We describe methods for the assay of hexosaminidase A and total hexosaminidase activities in dried blood spots (DBSs) on filter paper. MATERIALS AND METHODS: We studied 163 healthy controls, 9 Tay-Sachs patients, 4 Sandhoff patients, 18 obligate carriers and the newborn-screening cards from two patients with Tay Sachs and one patient with Sandhoff disease. To tubes containing a 3-mm-diameter blood spot, we added elution liquid and substrate solution. After incubation at 37 degrees C, the amount of hydrolyzed product was compared with a calibrator to allow the quantification of enzyme activity. RESULTS AND CONCLUSIONS: The described methodology is useful to distinguish patients with Tay-Sachs disease or Sandhoff disease from carriers and controls using samples that are sufficiently stable to be transported to the testing laboratory by mail. The diagnosis of both diseases from a newborn-screening card (NSC) was clearly demonstrated, even after storage for up to 38 months at room temperature. The newborn-screening card has been added to the biological materials that allow the identification of patients with Tay-Sachs disease and Sandhoff disease. PMID- 11880124 TI - Metabolic effects of C677T and A1298C mutations at the MTHFR gene in Brazilian children with neural tube defects. AB - BACKGROUND: Methylenetetrahydrofolate reductase (MTHFR) deficiency leads to impairment in folate metabolism and is implicated as a risk factor for neural tube defects (NTDs). Both C677T and A1298C MTHFR mutations are associated with NTDs, in some populations. METHODS: The frequencies of the C677T and A1298C MTHFR mutations were determined in 25 children with NTDs, case mothers and 75 healthy individuals from Sao Paulo City. Both C677T and A1298C mutations were analyzed by PCR-FLRP. The effects of MTHFR mutations on folate, vitamin B12 and homocysteine concentrations were also evaluated. RESULTS: C677T and A1298C allele frequencies in NTDs children and mothers were similar to that found in controls. Eleven in 23 NTDs patients and 10 in 21 NTDs mothers had folate or vitamin B12 concentrations in the lower end of the normal range. In NTDs children, C677T MTHFR genotypes did not affect vitamins and homocysteine concentrations, but plasma homocysteine was higher (p=0.028) in patients with 1298AA MTHFR genotype. Moreover, 677CT/1298AA haplotype was associated with lower vitamin B12 concentrations (p<0.05) in NTDs children. CONCLUSIONS: MTHFR gene mutations may affect vitamin B12 and homocysteine metabolism in Brazilian children with NTDs. PMID- 11880125 TI - A duality in the roles of reactive oxygen species with respect to bone metabolism. AB - BACKGROUND: Rampant production in skeletal abnormalities can lead to hyperoxidant stress though the production of "reactive oxygen species" (ROS) by osteoclasts, which assist in bone remodeling under physiological conditions. METHODS: Thirty cases each of post-menopausal osteoporosis, renal osteodystrophy and bone fractures constituted the test groups. Thirty healthy subjects made up the control group. Serum total alkaline phosphatase served as an index of osteoblastic activity. Serum calcium and phosphorous indicated bone remodeling status. Serum superoxide dismutase, glutathione peroxidase and glutathione reductase represented the enzymatic antioxidants. RESULTS: Mean values for malondialdehyde were significantly elevated (P<0.001) in test groups, indicating enhanced osteoclastic activity. Significantly depressed (P<0.001) activities of superoxide dismutase and glutathione peroxidase reinforced hyperoxidant stress. Mean values of glutathione reductase remained unaltered. Diminished osteoblastic activity in post-menopausal osteoporosis was indicated by depressed alkaline phosphatase (P<0.001). Increased serum calcium (P<0.001) and decreased serum phosphorous (P<0.001) in renal osteodystrophy indicated compensatory hyperparathyroidism. CONCLUSIONS: The findings indicate that ROS have a major role to play in bone metabolism. PMID- 11880126 TI - Uterine transplantation in humans: a new frontier. PMID- 11880127 TI - Transplantation of the human uterus. AB - Human uterine transplantation was performed on 6 April 2000 on a 26-year-old female who lost her uterus 6 years earlier due to post-partum hemorrhage. The donor, a 46-year-old patient with multiloculated ovarian cysts, underwent a hysterectomy modified to preserve tissue and vascular integrity. The donor uterus was connected in the orthotopic position to the recipient's vaginal vault and additional fixation was achieved by shortening the uterosacral ligament. The uterine arteries and veins were extended using reversed segments of the great saphenous vein, then connected to the external iliac arteries and veins, respectively. Immunosuppression was maintained by oral cyclosporine A (4 mg/kg/body wt.), azathioprine (1 mg/kg/body wt.) and prednisolone (0.2 mg/kg/body wt.). Allograft rejection was monitored by Echo-Doppler studies, magnetic resonance imaging (MRI), and measurement of the CD4/CD8 ratio in peripheral blood by fluorescence activated cell sorter (FACS scan). An episode of acute rejection was treated and controlled on the ninth day with anti-thymocytic globulin (ATG). The transplanted uterus responded well to combined estrogen--progesterone therapy, with endometrial proliferation up to 18 mm. The patient had two episodes of withdrawal bleeding upon cessation of the hormonal therapy. Unfortunately, she developed acute vascular thrombosis 99 days after transplantation, and hysterectomy was necessary. Macro- and microscopic histopathological examination revealed acute thrombosis in the vessels of the uterine body, with resulting infarction. Both fallopian tubes remained viable, however, with no evidence of rejection. The acute vascular occlusion appeared to be caused by inadequate uterine structure support, which led to probable tension, torsion, or kinking of the connected vascular uterine grafts. PMID- 11880128 TI - Serum creatine phosphokinase elevation in patients treated with intravenous magnesium sulfate. AB - OBJECTIVE: During the treatment of pre-term labor with magnesium sulfate, we noted an abnormal elevation of maternal serum creatine phosphokinase. This study was aimed at evaluating the relationship between tocolysis with MgSO4 and maternal serum CPK elevation, which represents the possible damage of muscles by magnesium sulfate. METHODS: Clinical records of 45 women treated with magnesium sulfate and beta-sympathomimetics for the treatment of pre-term labor were retrospectively examined. RESULTS: Serum CPK was abnormally elevated in 32 out of 45 cases (71.1%), but in only one out of 21 in the control group. In three cases, the decrease of serum creatine phosphokinase after cessation of magnesium sulfate was demonstrated, despite the continuous infusion of beta-sympathomimetics. CONCLUSION: Magnesium sulfate may cause muscular damage and abnormal elevation of maternal serum creatine phosphokinase. Special attention must be paid to patients when drugs acting on muscle cells, for example succinyl choline, are going to be used. PMID- 11880129 TI - Growth of the fetal lens and orbit. AB - OBJECTIVES: To present normal lens and orbital measurement ranges by gestational age. METHODS: Three hundred and thirty-five women with uneventful pregnancies were evaluated from the 15th to the 40th week of gestation. Lens and orbital measurements were added to routine biometric measurements for normal fetuses. RESULTS: A strong linear correlation was observed between gestational age (GA) and lens diameter (R(2)=0.89; P<0.0001), circumference (R(2)=0.89; P<0.0001), and surface (R(2)=0.90; P<0.0001). A linear correlation was also found between GA and orbital diameter (R(2)=0.92; P<0.0001), circumference (R(2)=0.92; P<0.0001) and surface (R(2)=0.95; P<0.0001). A linear growth function was observed between biparietal diameter and both lens diameter (R(2)=0.90; P<0.0001) and orbital diameter (R(2)=0.94; P<0.0001). CONCLUSION: Lens and orbital measurements provide normative data for fetal growth and development. These data may be also helpful in detecting fetal ocular anomalies. PMID- 11880130 TI - Effects of levonorgestrel implant upon TSH and LH levels in male infants during lactation. AB - OBJECTIVES: The objective of the study is to evaluate the effects of levonorgestrel transferred through breast milk on thyroid stimulating hormone (TSH) and luteinizing hormone (LH) levels in full breast-fed infants. METHODS: Forty healthy postpartum women and their male newborns were recruited for the study. Women were randomly allocated to two study groups: Group 1, IUD users and group 2, Norplant users. Blood and milk samples were collected on the day of IUD or Norplant insertion and 3 and 6 months later for TSH and levonorgestrel measurements. RESULTS: The results disclosed a significant decrease in TSH levels, and a negative relationship between LNG levels and TSH concentration in breast feeding infants at 3 months after implant insertion. The lowest TSH levels were observed at 6 months in the women from group 2. CONCLUSIONS: The overall data indicate that the LNG levels transferred to fully breast-fed infants through breast milk from Norplant users significantly modified their TSH levels. PMID- 11880131 TI - Tamoxifen-induced endometrial changes in postmenopausal women with breast carcinoma. AB - OBJECTIVES: To assess the effects of tamoxifen (TAM) on the endometrium in postmenopausal women. METHODS: A case control study of postmenopausal women with breast carcinoma, who were undergoing treatment in the Department of Radiotherapy and Surgery at the Christian Medical College Hospital, Vellore, India was done. Thirty-five women who were on tamoxifen (20 mg/day) for a period of at least 6 months formed the study group. Thirty-three women who were not receiving tamoxifen, formed the control group. Subjects in both groups had a pelvic examination and transvaginal sonogram followed by endometrial biopsy. RESULTS: There was a statistically significant difference in the mean endometrial thickness between the study group and control group (7.8+/-6.4 mm vs. 4.0+/-2.0 mm, respectively) More women in the tamoxifen group had an endometrial thickness of >5 mm but the number of women with polyps or hyperplasia of the endometrium did not differ significantly between the two groups. There were no women with endometrial carcinoma in either group. CONCLUSION: All patients on tamoxifen need to be evaluated by clinical examination annually. A transvaginal sonogram and endometrial biopsy/hysteroscopy may be performed on patients with abnormal vaginal bleeding, bloody discharge, staining or spotting. PMID- 11880132 TI - Bone-mass peak in multiparity and reduced risk of bone-fractures in menopause. AB - OBJECTIVES: Pregnancy has been proposed as a risk factor for the development of osteoporosis; however, the results are controversial. Women in Colombia are multiparous, and therefore they represent an ideal population for the investigation of the effect of parity on osteoporosis. METHODS: The study included 1855 post-menopausal Hispanic women from Barranquilla, Colombia who were referred to our osteoporosis clinic for a routine evaluation of their bone mineral status. Bone mineral density (BMD, g/cm(2)) was measured using dual energy X-ray absorptiometry. Total body, femoral and lumbar areas were scanned. RESULTS: BMD of total body, hip and legs (P<0.0001), Ward's area (P<0.002) and intertrochanteric area (P<0.003) was high in women with at least one delivery in comparison with nulliparous women. Total mineral and calcium body contents were also high in women after second delivery. A history of fractures was found in 22.9% (n=425) of the study women. The relative risk for bone-fractures was high (O.R. 0.41; P<0.000002) in nulliparous women in comparison to multiparous women. In addition, osteopenia (O.R. 2.01; P=0.008) and osteoporosis (O.R. 3.99; P=0.0004) were most often present in nulliparous women in comparison to multiparous women. CONCLUSIONS: Pregnancies in this population appear to be a protective factor against development of osteoporosis. The increase in total mineral and calcium contents was related to the number of pregnancies, suggesting the existence of a bone mass peak during pregnancy. PMID- 11880133 TI - Clinical value of early cleavage embryo. AB - OBJECTIVE: This study was undertaken to see if embryo transfer containing early cleavage embryos resulted in better clinical pregnancy rate. METHODS: The treatment outcomes of IVF-ET were retrospectively reviewed. Out of 258 transfer cycles, 160 cycles contained no early cleavage embryos (Group I) and 98 cycles contained at least one early cleavage embryo (Group II). The definition of early cleavage embryo is the presence of two blastomeres 24-26 h after insemination. The implantation rate, clinical pregnancy rate were compared between two groups. Student's t-test and the Mann-Whitney U-test were used for continuous variables, and the Chi-squared (chi(2)) test was used for binary variables. Differences were considered statistically significant at P<0.05. RESULT: The implantation rate and clinical pregnancy rate were 11.6% and 25.6% in Group I, 18.6% and 38.8% in Group II (P<0.05). CONCLUSION: Early cleavage embryos possess greater implantation potential. Embryo transfer containing early cleavage embryos had a better clinical pregnancy rate. PMID- 11880134 TI - Program note. Using UN process indicators to assess needs in emergency obstetric services. PMID- 11880136 TI - Maternal deaths associated with cesarean section in Enugu, Nigeria. AB - Maternal deaths from cesarean sections in Nigeria are exceptionally high and result from avoidable causes such as hemorrhagic shock, sepsis and hypertensive disorders in pregnancy. Increased involvement of specialists in the care and improved intra and post-operative management of cases are advocated to reduce the high maternal mortality rate. PMID- 11880137 TI - Nifedipine versus terbutaline in management of preterm labor. PMID- 11880138 TI - The epidemiology of group B streptococcal colonization among obstetrical and newborn populations in Kuwait. PMID- 11880139 TI - Comparison of IVF-ET outcomes under various GnRH agonist dosages in a long down regulation protocol. PMID- 11880140 TI - Ruptured corpus luteum cysts and anticoagulant therapy. PMID- 11880141 TI - Formal education does not improve the acceptance of cesarean section among pregnant Nigerian women. AB - OBJECTIVES: To investigate the relationship between formal education, personal beliefs and the acceptance of cesarean section among pregnant Nigerians. METHODS: A questionnaire survey was made of antenatal mothers seen over a 5-month period at a tertiary hospital in Enugu, Nigeria. RESULTS: A total of 1148 subjects were interviewed. Although as a single variable, formal education was significantly related to a more favorable attitude to cesarean section among the respondents, it ceased to be related when their beliefs about the operation were corrected for. CONCLUSIONS: Pregnant Nigerians' beliefs about cesarean section are more important than formal education in determining whether or not they accept it, hence the need to correct such false beliefs during antenatal classes. PMID- 11880142 TI - Deliveries at a district hospital in Rwanda, 1997-2000. AB - OBJECTIVES: To obtain data on pregnancy outcomes and maternal mortality at a district hospital in Rwanda. METHODS: All delivery records from January 1997 to December 2000 were reviewed. RESULTS: Over the 4-year period, 3408 women delivered a total of 3497 neonates, and 349 stillbirths (10%) occurred. The mean birthweight (twins excluded) was 3097 g and decreased significantly from 3160 g in 1997 to 3043 g in 2000. The prevalence of low birthweight was 12.5%. The mean age of women was 26.2 years. Cesarean section was done in 26% of deliveries at the hospital, equaling a cesarean section rate of 1.1% in the population. Nineteen maternal deaths occurred, yielding a (hospital) maternal mortality rate of 600 per 100000 live births. Uterine rupture occurred in 52 women, of whom six died (11.5%). CONCLUSIONS: Efforts to educate women at risk to deliver at a health center, and early referral of women to the hospital should be reinforced. The reduction of mean birthweight is of concern and reasons for this need to be analyzed. PMID- 11880143 TI - Golli-myelin basic proteins delineate the nerve distribution of lymphoid organs. AB - The golli-myelin basic proteins (MBPs) have been known to mark the nerve fiber extensions in both the peripheral nervous system (PNS) and the central nervous system. In this paper, we show that the nerve fibers revealed by neurofilament (NF) antibody staining in thymus and spleen, colocalized with golli in the capsular, trabecular (tr), and vasculature (v) systems. In the thymus, the density of these fibers was greater in the medulla than in the cortex. In the spleen, the golli immunoreactive fibers were seen within the capsule (ca), trabeculae, and along the artery tree, as well as the fine nerve fiber networks in the periarteriolar lymphoid sheath (PALS). Golli immunoreactivity appeared to colocalize with ER-TR7, a putative marker of connective tissue in lymphoid organs. However, further examination by Western blot analysis and immunohistochemistry performed on golli "knock out" mice showed that the antigens recognized by these two antibodies were different. The reason for the apparent colocalization of golli and ER-TR7 appears to be due to the close physical association of nerve fibers with connective tissue in these organs. These results suggest that golli immunoreactivity can visualize the distribution of nerve fibers in these lymphoid organs. PMID- 11880144 TI - Signaling pathways activated by chemokine receptor CXCR2 and AMPA-type glutamate receptors and involvement in granule cells survival. AB - We show that treatment of cerebellar granules with interleukin-8 (IL-8), growth related gene product beta (GRObeta) or AMPA induced activation of PI3-K/Akt and of ERK pathways, the latter being independent of PI3-K and dependent on PTX sensitive G proteins. We also show that AMPA-mediated neuron survival was abolished both by ERK kinase inhibitor PD98059 and AMPA-Rs blocker CNQX, and that chemokine-mediated survival was blocked by the PI3-K inhibitors LY294002 and wortmannin. We conclude that the neurotrophic effects of AMPA need the contemporary activation of ERKs and stimulation of AMPA-Rs, and that PI3-K/Akt activation is a determinant pathway for the IL-8/GRObeta anti-apoptotic activity. PMID- 11880145 TI - Myelin-specific tolerance attenuates the progression of a virus-induced demyelinating disease: implications for the treatment of MS. AB - Theiler's murine encephalomyelitis virus-induced demyelinating disease (TMEV IDD), a multiple sclerosis (MS) model, is a central nervous system (CNS) demyelinating disease characterized by early peripheral T cell responses to virus epitopes which spreads to myelin epitopes during chronic disease. We show that CD4(+) T cells isolated from the spinal cords of chronically infected SJL mice proliferate and secrete pro-inflammatory cytokines upon in vitro challenge with both TMEV epitopes and proteolipid protein (PLP(139-151)). Importantly, myelin specific tolerance induced by intravenous administration of MP4, a fusion of the myelin proteins myelin basic protein (MBP) and PLP, to SJL mice with ongoing TMEV IDD attenuated disease progression and resulted in significantly less demyelination and decreased inflammatory cell infiltration in the CNS. Paradoxically, peptide-specific splenic T cell proliferative and IFN-gamma responses were enhanced in the tolerized mice. Collectively, these results indicate that myelin-specific T cell responses contribute to chronic disease progression in this virus-induced model of MS, and suggest caution in the use of antigen-specific tolerance for treatment of ongoing autoimmune disease. PMID- 11880146 TI - Intramolecular epitope spreading induced by staphylococcal enterotoxin superantigen reactivation of experimental allergic encephalomyelitis. AB - The staphylococcal enterotoxin superantigens are among the most potent T cell stimulators known. They have been shown to alter the course of disease in experimental allergic encephalomyelitis, an animal model for multiple sclerosis (MS). We have previously demonstrated that two of the staphylococcal enterotoxins, SEA and SEB, are able to reactivate paralysis in PL/J mice which had been immunized with myelin basic protein (MBP) and resolved an initial episode of paralysis. In PL/J mice, Ac1-11 is the dominant encephalitogenic determinant of MBP. We hypothesized that superantigen reactivation of experimental allergic encephalomyelitis (EAE) may result in the spreading of T cell specificities for other epitopes of MBP. PL/J mice which had resolved an initial episode of EAE were treated with SEA and developed a second episode of paralysis. At the onset of symptoms, mice were sacrificed and splenocytes were stimulated in vitro with a panel of MBP peptides. EAE reactivation by SEA resulted in the spreading of T cell specificites to residues 100 to 120 of MBP. While intramolecular spreading did occur, spreading to other antigens did not, as evidenced by the lack of response to a proteolipid protein (PLP) peptide and heat shock protein 60 (hsp 60). To further characterize the epitope MBP 100-120, PL/J mice were immunized with MBP 100-120. No initial development of disease was observed. However, administration of SEA 2 weeks after MBP 100-120 immunization resulted in the onset of paralysis. In addition to a proliferative response to MBP 100-120, these mice also exhibited a proliferative response to the flanking MBP peptides 81-100 and 120-140. Thus, SEA is able to induce intramolecular epitope spreading in PL/J mice after reactivation of EAE. PMID- 11880147 TI - Failure of sensory neurons to express class I MHC is due to differential SOCS1 expression. AB - Neurons, unlike glia and most other cell types, fail to express major histocompatibility complex (MHC) molecules, even in response to IFNgamma. This study shows that sensory neurons but not glia constitutively express high levels of suppressor of cytokine signaling-1 (SOCS1), a negative regulator of IFNgamma signaling. Neurons from SOCS1 null mice expressed class I MHC in response to IFNgamma, while SOCS1 overexpression in glia inhibited MHC induction. Differential expression of SOCS1 by neurons explains their failure to express MHC. Regulation of SOCS1 induction and therefore neuronal responses to IFNgamma may be critical for neuron survival following viral infection or injury. PMID- 11880148 TI - Effects of staphylococcal enterotoxin A on pituitary-adrenal activation and neophobic behavior in the C57BL/6 mouse. AB - Bacterial superantigens, such as the staphylococcal enterotoxins (SE), exert a strong capacity for in vivo stimulation of T cell proliferation and cytokine production. Among these superantigens, staphylococcal enterotoxin B (SEB) has been shown to promote anxiety-like properties, possibly mediated by activation of central corticotropin-releasing hormone. In the present study, using male C57BL/6J mice, it was shown that challenge with another prominent superantigen, staphylococcal enterotoxin A (SEA), produced a dose-dependent (0.2-50 microg/mouse) increase in corticosterone and ACTH levels. Interestingly, while the adrenocorticoid response to SEA persisted in cyclosporine A-pretreated mice, it was completely abolished in RAG-1 deficient mice that lack functional T and B lymphocytes. The latter is consistent with the need for cellular interactions involving T cells and B cells (probably in an antigen-presenting capacity) that will initiate events leading to pituitary-adrenal activation by SEA. Since pituitary-adrenal activation typically alters "emotional" reactivity in animals, a final set of experiments assessed behavioral responses to an open field, exposure to a novel object, and a novel appetitive stimulus. These tests revealed a significant augmentation of reactivity to the novel object in SEA-challenged mice, although activity in the open field was not affected. Furthermore, consumption of a novel solution was reduced only if testing involved unfamiliar contextual circumstances. This suggested that anorexic effects per se were not induced by SEA at the dose used, but that attentional mechanisms focused on novelty were enhanced. PMID- 11880149 TI - Fas ligand mRNA expression in the mouse central nervous system. AB - Fas ligand (FasL) expressing cells delete Fas bearing T cells, thereby enabling privileged immune status in the brain. Although the presence of FasL immunoreactivity has been shown in various cell types in the central nervous system, the precise in vivo distribution of FasL mRNA in mammals is not known. Accordingly, we localized intense FasL mRNA signals in neuroglial cells mainly within the white matter regions. Using a combined labeling technique of immunocytochemistry and in situ hybridization, we confirmed that FasL signals were due to neuroglial cells rather than neurons. This study shows that FasL mRNA is constitutively expressed in the normal mouse brain, and suggests that the Fas/FasL system protects the CNS from immunological damage. PMID- 11880150 TI - NGF modulates CGRP synthesis in human B-lymphocytes: a possible anti-inflammatory action of NGF? AB - We investigated whether the sensory neuropeptide, calcitonin gene-related peptide (CGRP), could be synthesised by human lymphocytes. Our results indicate that in activated B-cells, there is a strong expression of CGRP gene transcripts, which is almost absent in resting cells. Since B-cells autocrinally produce NGF, the neutralisation of endogenous NGF by anti-NGF antibodies resulted in a marked reduction in CGRP expression in both resting and activated B-cells. Thus, NGF appears to directly affect the synthesis of CGRP in B-cells as in sensory neurons. By regulating CGRP synthesis in lymphocytes and neuronal cells, NGF can influence the intensity and duration of the immune response. PMID- 11880152 TI - Propagation of lewis rat encephalitogenic T cell lines: T-cell-growth-factor is superior to recombinant IL-2. AB - This study was designed to test the process of selecting encephalitogenic T cell lines in the Lewis rat using recombinant human IL-2 (rhIL-2) in comparison to TCGF. The lines were tested for growth, antigen induced proliferation, cytokine production, V-beta 8.2 expression and pathogenicity. We now report that rhIL-2 and TCGF were equally effective in supporting short-term pathogenic T-cell lines with similar proportion of V-beta 8.2 usage. For the maintenance of long term lines, however, TCGF was superior to IL-2. The concentration of rhIL-2 influenced the cultures: 10 units/ml led to more T-cell proliferation than either 2 or 50 units/ml. However, 50 units/ml of IL-2 led to enhanced Th1 polarization. Thus, the type and concentration of growth factors can influence both the propagation of T cells and their phenotype. PMID- 11880151 TI - Regulation of calcium currents by chemokines and their receptors. AB - We investigated the modulation of voltage dependent Ca(2+) currents by chemokine receptors in heterologous expression systems and neurons. Fractalkine, SDF 1alpha, RANTES and MDC inhibited the I(Ba) in CX3CR1-, CXCR4-, CCR5- and CCR4 expressing G1A1 cells, respectively. The I(Ba) inhibition was voltage-dependent, exhibited prepulse facilitation, and was blocked by N-ethylmaleimide and pertussis toxin pretreatment, indicating that it was mediated by Gi/Go. Some chemokines also inhibited the I(Ba) in subpopulations of dorsal root ganglion neurons and area postrema/nucleus tractus solitarius neurons. These data provide evidence that chemokines can potentially modulate neuronal signaling through the inhibition of neuronal Ca(2+) currents. PMID- 11880153 TI - Calcitonin gene-related peptide induces AP-1 activity by a PKA and c-fos dependent mechanism in pre-B cells. AB - Calcitonin gene-related peptide (CGRP) is a sensory neuropeptide with regulatory influences on immune and inflammatory responses and early B lymphocyte differentiation. Little is known about its cellular mechanisms. These studies examined whether CGRP induces c-fos. CGRP induced a transient concentration dependent increase in c-fos in a CGRP receptor expressing pre-B cell line, 70Z/3. CGRP did not induce c-jun, jun B or jun D. A protein kinase A (PKA) inhibitor blocked c-fos induction by CGRP, suggesting that the mechanism depends on cAMP induction of PKA. CGRP induced AP-1 binding activity in the nucleus, indicating that CGRP regulates expression of specific target genes. CGRP also induced c-fos in B220(+) enriched cells from bone marrow. These results suggest that regulatory influences of CGRP on early B cells and in other tissues can involve a cAMP/PKA, c-fos/AP-1-dependent pathway for regulation of specific genes. PMID- 11880154 TI - Constitutive expression of proinflammatory complement components by subsets of neurons in the central nervous system. AB - The brain is largely protected from damage due to infection, trauma, and aberrant processes by the innate immune system. These studies were undertaken to determine whether neurons in normal brains constitutively express complement components. In situ hybridization and immunohistochemical studies with specific riboprobes and antibodies, respectively, revealed that most hippocampal neurons, many pyramidal cortical neurons and cerebellar Purkinje neurons in normal murine brains constitutively express C3, C5 and C6. The constitutive expression by neuronal subsets of components of the complement activation and membrane attack pathways suggests that the complement system represents a "first line" of host defense in the brain. PMID- 11880155 TI - Cholinergic and serotonergic activities are required in triggering conditioned NK cell response. AB - The purpose of the study was to examine the importance of the cholinergic system in triggering the conditioned NK cell response. The fact that serotonergic system can modulate cholinergic functions suggested that it might be involved in conditioned NK cell response. To evaluate the potential pathways, cholinergic and serotonergic antagonists were applied centrally at either the conditioned association or recall stage, to interfere with the conditioned NK cell response. The results showed that both the cholinergic and serotonergic systems were necessary for eliciting the conditioned enhancement of NK cell activity. Involvements of the two systems were found to be critical for establishing the conditioned association and recall of the conditioned response. The blocks are believed to be receptor mediated. The receptors identified to be involved in the regulation of the conditioned NK cell response were: M(1), M(2) and M(3) muscarinic; nicotinic; 5 HT(1) and 5 HT(2) receptors. PMID- 11880156 TI - Activation of Vbeta8 T cells affects spontaneous EAE in MBP TCR transgenic mice. AB - Two strains of transgenic (Tg) mice (Valpha2.3/Vbeta8.2 and Valpha4/Vbeta8.2) have T cell receptors (TCR) that recognize the NAc1-11 immunodominant epitope of the myelin basic protein (MBP). Spontaneous experimental autoimmune encephalomyelitis (sEAE) readily develops in Valpha2.3/Vbeta8.2 mice. T cells in Valpha2.3/Vbeta8.2 mice demonstrate increased levels of CD69, CD44(high) and decreased CD45RB relative to Valpha4/Vbeta8.2 mice. Increased proliferative responses to MBP and high levels of TNF-alpha are seen in Valpha2.3/Vbeta8.2 mice. High IL-4 and TGF-beta production is observed in Valpha4/Vbeta8.2 mice. CC chemokines (macrophage inflammatory protein-1 alpha (MIP-1alpha), RANTES and monocyte chemotactic protein 1 (MCP-1)) are increased in the central nervous system (CNS) of Valpha2.3/Vbeta8.2 mice. Thus, activated Th1 cells in the periphery of Valpha2.3/Vbeta8.2 mice may traffic to the CNS in response to CC chemokines, influencing sEAE. PMID- 11880157 TI - Adoptive protection from experimental myasthenia gravis with T cells from mice treated nasally with acetylcholine receptor epitopes. AB - Nasal administration of synthetic CD4(+) epitopes of the acetylcholine receptor (AChR) prevents experimental myasthenia gravis (EMG) in C57Bl/6 mice, but not in IL4-deficient C57Bl/6 (IL4(-/-)) mice. Here we verify that nasal tolerance requires IL4, by showing that CD4(+) cells from C57Bl/6 mice treated nasally with a pool of AChR CD4(+) epitopes protected IL4(-/-) mice from EMG and caused a reduced production of anti-AChR antibody. CD4(+) cells from C57Bl/6 mice treated with unrelated peptides or sham-treated did not induce protection. CD4(+) cells from C57Bl/6 mice treated with just one AChR peptide protected IL4(-/-) mice from EMG without affecting antibody synthesis. PMID- 11880158 TI - Enhanced expression of iNOS intratumorally and at the immunization site after immunization with IFNgamma-secreting rat glioma cells. AB - Nitric oxide (NO) can modulate both tumor growth and antitumor immune responses. In order to elucidate the mechanism of curative therapeutic immunization with IFNgamma-producing glioma cells, we examined the expression of inducible nitric oxide synthase (iNOS) in tissue sections from immunized animals. There was a significantly enhanced iNOS expression both intratumorally and at the immunization site. Although the mechanisms behind this dual expression of iNOS most probably are different, our results suggest a role for NO in both the induction and execution of the antitumor response. PMID- 11880159 TI - Identifying disease modifying genes in multiple sclerosis. AB - Evidence is mounting that genetic variation influences not only susceptibility to multiple sclerosis (MS), but also its course and severity. Identification of disease modifying genes, however, poses unique challenges, especially on how to classify the course and outcome of the disease in ways that may be relevant to analysis of biological factors that might be influenced by genes. The power of the statistical approaches to detect small effects of individual genes in complex disorders such as MS is problematic, and approaches to estimate power must be appropriate for the data. Nonetheless, using contemporary schemes of classification, genetic variants that influence disease course have been found; in fact, a small number have been confirmed to influence disease course in two or more independent studies. This review addresses strategies relevant to identification of disease modifying genes in MS, and summarizes and critically evaluates the current state of knowledge in this area. PMID- 11880160 TI - Interferon-beta increases the stimulatory capacity of monocyte-derived dendritic cells to induce IL-13, IL-5 and IL-10 in autologous T-cells. AB - Dendritic cells (DCs) are key regulators of immune responses and have been associated with autoimmunity in animal models and human disease. The effects of interferon beta (IFN-beta), an immunomodulatory cytokine used in multiple sclerosis (MS) therapy, on DCs are not well understood. Monocyte-derived DCs at different stages of maturation were stimulated with IFN-beta and DC-phenotype and stimulatory function were measured. IFN-beta inhibited the development of DCs at early stages but enhanced DC maturation. Moreover, IFN-beta enhanced the capacity of DCs to stimulate autologous T-cells to secrete IL-13, IL-10 and IL-5. Thus, IFN-beta has both immunostimulatory and immunosuppressive effects on DCs depending on the stage of maturation. PMID- 11880161 TI - Interferon beta-1b modulates MCP-1 expression and production in relapsing remitting multiple sclerosis. AB - Monocyte chemoattractant protein-1 (MCP-1) seems to be involved in the pathogenesis of multiple sclerosis (MS). We found that in unstimulated (PHA(-)) and PHA-stimulated (PHA(+)) peripheral blood mononuclear cells (PBMC), MCP-1 and TNFalpha levels are higher in stable untreated MS patients. Interferon gamma (IFNgamma) is higher in relapsing patients in PHA(-) cultures and in stable patients in PHA(+) cultures. Chronic IFNbeta-1b treatment down-regulates TNFalpha, IFNgamma and MCP-1 production except for TNFalpha in relapsing patients. IFNbeta-1b, in vitro, increases MCP-1, TNFalpha and IFNgamma spontaneous production in all patients. Multivariate analysis suggests that MCP-1 production is dependent from clinical status and not from TNFalpha and IFNgamma production. Logistic regression analysis shows that MCP-1 production is significantly modified by treatment. Further studies are needed to clarify the role of MCP-1 in MS. PMID- 11880162 TI - Human lymphocyte growth hormone stimulates interferon gamma production and is inhibited by cortisol and norepinephrine. AB - Lymphocyte growth hormone (L-GH) is distributed throughout the human immune system; however, its biologic role has not been defined. In order to clarify this issue, we determined if Candida, commonly used as a recall antigen, and IL-12, an important monocyte cytokine that stimulates the TH1 cytokine pathway, could stimulate L-GH mRNA synthesis in cultured human peripheral blood mononuclear cells (PBMCs). Both Candida and IL-12 produced significant (p<0.01) stimulation of L-GH mRNA synthesis and the TH1 cytokine, gamma interferon (IFN-gamma). These effects on IFN-gamma production were significantly (p<0.001) inhibited by a GH antagonist, suggesting that L-GH was acting by an autocrine or paracrine mechanism to enhance IFN-gamma production in these serum-free cultured cells. Concentrations of norepinephrine and cortisol achievable in humans during stress were able to significantly (p<0.001) decrease L-GH synthesis in the PBMCs. They also diminished IFN-gamma significantly (p<0.001), but not the TH2 cytokine, IL 10, in the supernatants of the cultured PBMLs. These studies suggest that L-GH plays a role in cellular immune function mediated via the TH-1 pathway. PMID- 11880163 TI - Treatment with glatiramer acetate induces specific IgG4 antibodies in multiple sclerosis patients. AB - We analysed the humoral immune response to glatiramer acetate (GA, Copaxone) in 20 multiple sclerosis patients treated with GA, 20 patients not treated with GA and 20 normal control subjects. Using an ELISA for detection of total GA-reactive immunoglobulins (all isotypes), all treated patients but also 3/20 untreated and 8/20 healthy subjects scored positive at 1:20 plasma dilutions. At higher dilutions, 5/20 treated patients and two healthy donors had relatively high levels of anti-GA antibodies. Isotype and IgG subclass analysis revealed that the two antibody-positive normal subjects had IgM and small titers of IgG1 or IgG2 antibodies. In contrast, 18 of 20 GA-treated patients, had low but significant titers of GA-reactive IgG4 antibodies. This finding is consistent with the previously described GA-mediated induction of T-helper 2 (TH2)-like regulatory T cells. PMID- 11880164 TI - No evidence to support CTLA-4 as a susceptibility gene in MS families: the Canadian Collaborative Study. AB - Two polymorphisms of the CTLA-4 gene were genotyped in 232 sibling pairs affected with multiple sclerosis (MS) from 185 families. The CTLA-4 polymorphisms genotyped were a 3' untranslated (AT)(n) microsatellite and an alanine/threonine RFLP of exon 1. There was no evidence observed for linkage by either identity-by descent (ibd) or identity-by-state (ibs) methods. A transmission disequilibrium test (TDT) was performed and no preferential transmission of alleles was observed. Upon stratification of patients, there was no preferential transmission observed based upon gender, by presence or absence of HLA*DRB1*15, by ethnicity or by clinical course of the disease. CTLA-4 does not appear to be a major MS susceptibility locus in Canadian multiplex families. PMID- 11880165 TI - [Pelvis obliquity, hip excentration and scoliosis in a population of 120 polyhandicaped adults. Descriptive study]. AB - The objective of this study is to give objective data about neuro-orthopaedic status of the population of polyhandicapped patients for improving both our medical and surgical practices. MATERIAL AND METHOD: It was a multicentre, cross sectional study to describe the population of polyhandicapped patients over 15 years of age. Each person was provided with clinical examination and X rays to detect scoliosis, pelvis obliquity and hip excentration (subluxation or dislocation) and other factors associated with polyhandicap. Polyhandicap was defined as a severe handicap linking physical and mental disability and responsible for extreme restriction of autonomy. RESULTS: One hundred and twelve persons were studied. Mean age was 27.2 years. Etiology of the handicap was neonatal asphyxia (38.3% of patients). 69.0% of patients had no possibility to turn over in lying position. Orthopaedic status was serious : 75.9% scoliosis, 65.2% pelvis obliquity (75% left and 25% right), 19.1% hip dislocation. General condition was precarious : one patient in two had a weight judged lower than normal weight, 32.5% had bedsores, 45.2% suffered from hip pain and only 13.6% received treatment against pain. DISCUSSION AND CONCLUSION: The description of general and neuro-orthopaedic status of these 120 people with polyhandicap cannot leave us indifferent. Although life expectancy in this population was lower than in normal populations, we are faced with life process and no with accompanying the terminally ill. We urgently need to review preventive care in terms of quality and quantity as well as surgical proposals for treating spasticity in children and adults. PMID- 11880166 TI - [Techniques for repair of chondral and osteochondral lesions in the knee. Principles of rehabilitation]. AB - The objective of the present study is to assess the techniques, indications and management of the three principal repair methods of cartilaginous lesions of the knee. Arthroscopic irrigation and debridement are excluded. The most important articles of the last 20 years have been evaluated favouring those written by the protagonists of the technique and those with the longest follow-up. Each technique, i.e. the microfractures, the osteochondral grafts and the chondrocyte grafts, is explained in detail with its surgical principles, management of specific reeducation and results. Postoperative treatment is not very different during the first weeks; for condyle lesions protection of the graft site is mandatory by strict avoidance of non weight bearing, whereas immediate mobilisation of the knee is possible; for patellar lesions, weight bearing is permitted whereas flexion is limited. Later, the delay concerning resumption of weight bearing, muscular training and sports is specific for each technique. These three techniques are only apply to symptomatic lesions of the condyle at the weight-hearing zone or the femoro-patellar site in young patients. The degenerative knee, even in its beginning, is excluded. Those lesions can be linked to rupture of the anterior cruciate ligament and immediate cartilaginous repair is discussed. PMID- 11880168 TI - [How mirror feedback improves undisturbed upright stance control]. AB - OBJECTIVE: To evaluate, in normal and healthy adults, the postural effects induced by a protocol of body sway mirror feedback. MATERIALS AND METHODS: Two experimental conditions, one consisting in staring eyes open (EO) and one consisting in mirror feedback (MFB), are randomly proposed. Displacements of the centre of pressure (CP), issued from a force platform, are advantageously decomposed along medio-lateral ML and antero-posterior AP directions into two elementary components: the horizontal motions of the centre of gravity (CG(h)) and the difference CP-CG(v). The former is recognised as the main controlled variable in upright stance whilst several clues can be assessed by the latter: the overall muscular activity, the ankle stiffness and the horizontal accelerations communicated to the CG. Elementary motions CG(h) and CP-CG(v) are then processed in the frequential domain and characterized through RMS and MPF parameters. RESULTS: For CP-CG(v) motions the MFB protocol induces larger MPF along the AP direction and thus an increased ankle stiffness. In addition, the RMS are slightly decreased, indicating that less motor units are called into play. On the other hand, no real effect is observed in the ML direction. A general trend is that the CG(h) amplitudes are diminished by the MFB protocol, especially in the ML direction where a statistical effect is noticed. CONCLUSION: The specificity of the effects induced by the MFB technique makes it an appropriate tool for re-establishing, especially on the AP direction, the balance functions, in particular those involving exaggerated CP-CG(v) motions. PMID- 11880167 TI - [Hemiplegia and return to domicile]. AB - OBJECTIVE: We have studied the outcome after a stroke on being discharged from a department of physical medicine and rehabilitation in patients with hemiplegia, and the factors influencing the choice of the orientation. MATERIAL AND METHOD: This prospective study was based on 93 patients. The mode of discharged was patient's domicile, institution, the number of death was also noted. The potentially influential factors studied were age, the side with hemiplegia, the aetiology of the hemiplegia, co-morbidity, the delay in starting rehabilitation, the neurological damage evaluated by the Orgogozo score, the initial functional damage evaluated by Functional Independence Measure (FIM) and by the functional score carried out within the framework PMSI(1) (computerised programme of medical care), the existence of aphasia, the existence of a depressive syndrome, presence of hemineglect, presence of superficial or profound sensory disorders, incontinence at the start of rehabilitation and at one month after the stroke, the existence of cognitive or psychiatric disorders. The tests used were the non parametric test of Mann and Whitney, the chi(2) test and the correlation test. The threshold of significance used was 0.05. RESULTS: Based on 93 patients (47 women and 46 men, average age 64.8) 81 have gone back to their previous domicile, 11 were oriented toward an institution. One patient died. The predictive factors or those linked to an absence of return to the previous domicile were the age, social situation, the delay in starting rehabilitation, presence of aphasia, the initial and final functional damage, the impossibility to walk, the presence of a depressive syndrome, urinary incontinence. CONCLUSION: The authors stress the importance of familial environment and of functional independence in establishing a prognosis for return to the domicile. PMID- 11880169 TI - A note on the quantitative properties of McGhee-von Hippel model. AB - Exact closed-form expressions are presented for the properties of the McGhee-von Hippel model of non-specific binding of large ligands to one-dimensional homogeneous lattices. These properties include the midpoint location and the slope at the middle point of the binding isotherms (v varies with ln L plots), the location and magnitude of the maximum, as well as the location of the inflection point, in the Scatchard plots (v/L varies with v plots). PMID- 11880170 TI - Lateral distribution of cholesterol in membranes probed by means of a pyrene labelled cholesterol: effects of acyl chain unsaturation. AB - The lateral distribution of cholesterol in membranes in the fluid state was investigated by studying the variation of the molar absorption coefficient of pyrene-labelled cholesterol (Py-chol) vs. its concentration in vesicles made of phosphatidylcholine, with variable acyl chain unsaturations. Absorption measurements indicated non-ideal mixing of Py-chol in unsaturated lipids, a process mainly controlled by the cholesterol moiety of the probe. Similar abilities of cholesterol and Py-chol in perturbing the phase properties of pure saturated phosphatidylcholine were observed by DSC experiments. Immiscibility of sterols was corroborated by fluorescence polarization measurements, which indicated a weaker ordering effect of cholesterol in unsaturated membranes. The sizes and the quantities of sterol oligomers formed were calculated. A model for the lateral distribution of cholesterol in membranes is proposed and is applied to known cholesterol/phosphatidylcholine phase diagrams. Finally, the results are discussed with regard to recent models of biological membrane organization, (i.e. rafts). PMID- 11880171 TI - From the sequence to the superstructural properties of DNAs. AB - A theoretical model for predicting intrinsic and induced DNA superstructures as well as their thermodynamic properties is presented. Intrinsic sequence-dependent superstructures are evaluated by integrating local deviations from the canonical B-DNA of the different dinucleotide steps. Induced superstructures are obtained by adopting the principle of minimum deformation free energy, evaluated in the Fourier space, in the framework of first-order elasticity. Finally dinucleotide stacking energies and melting temperatures are considered to account for local flexibility. In fact the two scales are strongly correlated. The model works very satisfactorily in predicting the sequence-dependent effects on the DNA experimental behavior, such as the gel electrophoresis retardation, the writhe transitions in topologically constrained domains, the thermodynamic constants of circularization reactions as well as the nucleosome thermodynamic stability constants. PMID- 11880172 TI - Correlative motions and memory effects in molecular dynamics simulations of molecules: principal components and rescaled range analysis suggest that the motions of native BPTI are more correlated than those of its mutants. AB - In this work MD simulations of the native bovine pancreatic trypsin inhibitor (BPTI) and 16 mutants were done in vacuum in order to study memory effects in the mutants using principal component analysis (PCA) and the rescaled range analysis (Hurst exponents). Both PCA and the rescaled range analysis support our previous proposition, based on PCA of lysozyme, that the motions of a native protein are more correlated than those of mutants. The methods are compared, the nature and applications of the rule and the role of the long-range correlations in MD time series (i.e. memory) are discussed in the context of collective motions. PMID- 11880173 TI - A new approximate whole boundary solution of the Lamm differential equation for the analysis of sedimentation velocity experiments. AB - Sedimentation velocity is one of the best-suited physical methods for determining the size and shape of macromolecular substances or their complexes in the range from 1 to several thousand kDa. The moving boundary in sedimentation velocity runs can be described by the Lamm differential equation. Fitting of suitable model functions or solutions of the Lamm equation to the moving boundary is used to obtain directly sedimentation and diffusion coefficients, thus allowing quick determination of size, shape and other parameters of macromolecules. Here we present a new approximate whole boundary solution of the Lamm equation that simultaneously allows the specification of sedimentation and diffusion coefficients with deviations smaller than 1% from the expected values. PMID- 11880174 TI - X-Ray analysis of d(CGCGAATTXGCG)(2) containing a 2' or minute-deoxy-N(4) methoxycytosine residue at X: a characteristic pattern of sugar puckers in the crystalline state of the Dickerson-Drew type DNA dodecamers. AB - In a series of structural studies on damaged DNA, a modified Dickerson-Drew dodecamer with the sequence d(CGCGAATTmo(4)CGCG), where mo(4)C is 2'-deoxy-N(4) methoxycytidine, was synthesized and its structure in a new crystal form has been determined by the X-ray diffraction method. The two dodecamers form a B-form duplex, in which the two mo(4)C residues, respectively, form a wobble pair and a Watson-Crick type pair with the guanine residues of the opposite strand. A comparison of the sugar conformations with those of the other related Dickerson Drew dodecamers indicates a common feature of their puckering patterns. The sugar pucker of the third residue always adopts an intermediate state (C4'-exo-O4' endo) between the A-form and B-form. This deviation is ascribed to the stacking interaction of the ribose ring at the third residue with the guanine base at the 12th residue, which is brought about by an extra G12:G2 interaction between two duplexes related by a crystallographic 2(1) symmetry. PMID- 11880176 TI - Oncogenic functions of tumour suppressor p21(Waf1/Cip1/Sdi1): association with cell senescence and tumour-promoting activities of stromal fibroblasts. AB - p21(Waf1/Cip1/Sdi1) is best known as a broad-specificity inhibitor of cyclin/cyclin-dependent kinase complexes, but p21 also interacts with many other regulators of transcription or signal transduction. p21 induction, which is mediated by p53 and by p53-independent mechanisms, is essential for the onset of cell cycle arrest in damage response and cell senescence. The effects of p21 knockout in mice and its expression patterns in human cancer are consistent with a role for p21 as both a tumour suppressor and an oncogene. Several functions of p21 are likely to promote carcinogenesis and tumour progression. These include endoreduplication and abnormal mitosis that develop in tumour cells after release from p21-induced growth arrest, the ability of p21 to inhibit apoptosis through several different mechanisms, and its ability to stimulate transcription of secreted factors with mitogenic and anti-apoptotic activities. The latter effects of p21 show close resemblance to paracrine activities of senescent cells and to tumour-promoting functions of stromal fibroblasts. Therapeutic strategies targeting the oncogenic consequences of p21 expression may provide a new approach to chemoprevention and treatment of cancer. PMID- 11880175 TI - Modulation of cyclin-dependent kinase 4 by binding of magnesium (II) and manganese (II). AB - All kinases require an essential divalent metal for their activity. In this study, we investigated the metal dependence of cyclin-dependent kinase 4 (CDK4). With Mg(2+) as the essential metal and MgATP being the variable substrate, the maximum velocity, V, was not affected by changes in metal concentration, whereas V/K was perturbed, indicating that the metal effects were mainly derived from a change in the K(m) for MgATP. Analysis of the metal dependence of initial rates according to a simple metal binding model indicated the presence on enzyme of one activating metal-binding site with a dissociation constant, K(d(a)), of 5 +/-1 mM, and three inhibitory metal-binding sites with an averaged dissociation constant, K(d(i)), of 12+/-1 mM and that the binding of metal to the activating and inhibitory sites appeared to be ordered with binding of metal to the activating site first. Substitution of Mn(2+) for Mg(2+) yielded similar metal dependence kinetics with a value of 1.0+/-0.1 and 4.7+/-0.1 for K(d(a)) and K(d(i)), respectively. The inhibition constants for the inhibition of CDK4 by MgADP and a small molecule inhibitor were also perturbed by Mg(2+). K(d(a)) values estimated from the metal variation of the inhibition of CDK4 by MgADP (6+/ 3 mM) and a small molecule inhibitor (3+/-1 mM), were in good agreement with the K(d(a)) value (5+/-1 mM) obtained from the metal variation of the initial rate of CDK4. By using the van't Hoff plot, the temperature dependence of K(d(a)) and K(d(i)) yielded an enthalpy of -6.0 +/- 1.1 kcal/mol for binding of Mg(2+) to the activating site and -3.2 +/- 0.6 kcal/mol for Mg(2+) binding to the inhibitory sites. The values of associated entropy were also negative, indicating that these metal binding reactions were entirely enthalpy-driven. These data were consistent with metal binding to multiple sites on CDK4 that perturbs the enzyme structure, modulates the enzyme activity, and alters the affinities of inhibitor for the metal-bound enzyme species. However, the affinities of small molecule inhibitors for CDK4 were not affected by the change of metal from Mg(2+) to Mn(2+), suggesting that the structures of enzyme-Mg(2+) and enzyme-Mn(2+) were similar. PMID- 11880177 TI - Role of oxidative DNA damage caused by carbon tetrachloride-induced liver injury - enhancement of MeIQ-induced glutathione S-transferase placental form-positive foci in rats. AB - The strong association between chronic inflammation and development of cancer is well-established in chronic inflammatory states. Nitric oxide (NO) is generated by inflammatory cytokines due to the action of inducible nitric oxide (iNOS), oxidizing DNA to form 8-hydroxy-2'-deoxyguanosine (8-OHdG) adducts, a major species of oxidative DNA damage. In the present study, we investigated the enhancing effect of carbon tetrachloride, a typical hepatotoxic chemical, on rat 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx) hepato-carcinogenesis. A total of 420, 21-day-old, male Fisher 344 rats were given MeIQx at a concentration of 0, 0.001 ppm (human exposure level), 0.01, 0.1, 1, 10 and 100 ppm in the diet, and each group was separated into carbon tetrachloride-treated and vehicle-treated subgroups. Carbon tetrachloride was given by subcutaneous (s.c.) injection twice a week at a dose of 0.125 ml/kg body weight (b.w.) for the first 10 weeks and then at 0.25 ml/kg b.w. during the next 10 weeks. All rats were sacrificed at the end of week 22. In the vehicle-treated animals, only 100 ppm MeIQx significantly increased the number of glutathione S-transferase placental form (GST-P)-positive foci in the liver compared with 0 ppm MeIQx. Co administration of carbon tetrachloride enhanced the induction of GST-P-positive foci by MeIQx in each group and the curve was almost the same pattern as that of vehicle-treated group but their numbers were significantly enhanced with 10 ppm and above compared with 0 ppm MeIQx. Persistent liver injury and liver cell proliferation were histopathologically observed in carbon tetrachloride-treated groups. Increase of 8-hydroxydeoxyguanosine (8-OHdG) formation and iNOS overexpression were observed by co-administration of carbon tetrachloride in MeIQx-treated rat liver. Our results indicate that carbon tetrachloride enhances MeIQx hepato-carcinogenicity through increase in oxidative DNA damage but non effect levels of MeIQx carcinogenic activity still exist. PMID- 11880178 TI - Cellular thiol status-dependent inhibition of tumor cell growth via modulation of retinoblastoma protein phosphorylation by (-)-epigallocatechin. AB - Tea polyphenols have been shown to inhibit tumor cell growth, but there is limited information on their effects on cell signaling and cell cycle control pathways. We have shown the involvement of such mechanisms as activation of mitogenic activated protein kinases, decreases in ornithine decarboxylase activity and in cellular thiol levels, elicitation of mitochondrial cytochrome c release, and activation of caspases by the green tea galloyl polyphenol, epigallocatechin (EGC). In the current study, we sought to determine how EGC alters cell cycle and its related control factors in its growth inhibitory effect in Ehrlich ascites tumor cells. The significant finding here is that EGC caused a dose-dependent accumulation of cells in the G1 phase and a decrease in the phosphorylation of the retinoblastoma (Rb) protein, which was also in a cellular thiol-dependent manner. The involvement of a cellular thiol-dependent modulation in Rb phosphorylation leading to the regulation of tumor cell growth by a green tea polyphenol is a novel observation, to the best of our knowledge. PMID- 11880179 TI - Down-regulation of von Hippel-Lindau protein in N-nitroso compound-induced rat non-clear cell renal tumors. AB - Non-clear cell rat kidney tumors, inducible by N-nitroso compounds but lacking mutations in the von Hippel--Lindau (VHL) coding sequence, were examined for other VHL alterations. Neither mutations nor DNA methylation was detected in a putative promoter region. By immunohistochemistry, however, VHL protein level was evidently reduced in six of the eight eosinophilic renal epithelial tumors and in all the ten nephroblastomas. Immunoblotting of normal kidney detected two VHL proteins of 20 and 22kDa in a 16-day-old fetal rat but only 20kDa protein in an adult rat. This is the first demonstration of VHL alteration at the protein level. PMID- 11880180 TI - Perinatal selenium exposure decreases spontaneous liver tumorogenesis in CBA mice. AB - This study was designed to investigate the influence of neonatal selenium (an anticarcinogenic agent) exposure on spontaneous liver tumor formation in adult mice. Pregnant CBA mice were administered selenium during the last week of pregnancy and for ten days following parturition. Selenium significantly reduced the incidence of spontaneous hepatomas in adult male progeny, while having no effect on the lower hepatoma incidence in adult females. The data indicate that neonatal selenium alters hepatoma incidence in a sex-dependent manner. This study represents the model of neonatal cancer prevention. PMID- 11880182 TI - Methionine restriction induces apoptosis of prostate cancer cells via the c-Jun N terminal kinase-mediated signaling pathway. AB - Tumors are relatively more sensitive to methionine restriction than corresponding normal tissues, a phenomenon known as methionine auxotrophy. The current studies were undertaken to elucidate the molecular mechanisms for methionine auxotrophy of prostate cancer cells. We found that the activity of c-Jun N-terminal kinase 1 (JNK1) increased dramatically in response to methionine restriction. Over expression of wild type JNK1 by transient transfection enhanced apoptosis in response to methionine restriction, whereas over expression of a kinase inactive mutant of JNK1 protected PC-3 human prostate cancer cells from apoptosis. We conclude that JNK1 plays a critical role in signaling cancer cells to undergo apoptosis in response to methionine restriction. PMID- 11880181 TI - Photodynamic therapy of human breast cancer xenografts lacking caspase-3. AB - The human breast cancer cell line MCF-7 is deficient in procaspase-3 and in caspase-3-dependent steps in apoptosis due to deletion of the CASP-3 gene. We previously found that the cells transfected with empty vector (MCF-7v cells) were considerably less sensitive to photodynamic treatment in vitro with the phthalocyanine photosensitizer Pc 4 than were the cells stably transfected with human procaspase-3 cDNA (MCF-7c3 cells); however, overall cell killing, as determined by a clonogenic assay, was not affected by the presence of procaspase 3. The present study was undertaken to determine whether photodynamic therapy (PDT) in vivo was dependent on the ability of the cells to carry out the late steps in apoptosis that are catalyzed by this caspase. Xenografts of MCF-7 cells and the isogenic-derived MCF-7v and MCF-7c3 cells were generated in female athymic nude mice implanted with an estrogen pellet. MCF-7c3 xenografts, but not those of the other two lines, continued to express procaspase-3, as revealed by Western blots of proteins from the cells and the xenografts. When the xenografts reached 50-120 mm(3), some were treated with PDT (1mg/kg Pc 4 i.v. followed 48 h later by 150 J/cm(2) light at 672 nm and 150 mW/cm(2)), while others served as controls (no treatment, light alone, or Pc 4 alone). All Pc 4-PDT-treated tumors and none of the controls exhibited either complete or strong partial responses, and complete responses were durable for the entire observation period of 16 days. The responses were not dependent upon the presence of procaspase-3 in the xenografts. The results indicate that the rapid response of Pc 4-PDT-treated tumors in vivo is not due to their ability to carry out the major caspase-3 mediated late steps in apoptosis. PMID- 11880183 TI - Importance, mechanisms and limitations of the distant bystander effect in cancer gene therapy of experimental liver tumors. AB - GCV-ablation of transplanted TK-positive liver tumors or the application of syngenic and allogenic HSV-TK/GCV oncolysates significantly reduced the size of synchronously growing untreated sister tumors in the liver. These TK-negative liver tumors constantly showed an increased infiltration by mononuclears (x4). The relative abundance of CD 4/8, NK and monocyte subtypes remained constant. The distant bystander effect was associated with a strong induction of GMCSF and IL 12 expression in the untreated TK-negative liver tumors. Analysis of the vbeta T cell receptor profiles from TK-negative tumors did not point to clonal lymphocyte expansions. These results support the view of the 'distant bystander effect' as a predominantly local phenomenon, which is mediated by resident immune effectors rather than by MHC I restricted CD 3 positive lymphocytes. PMID- 11880184 TI - CASK and its target gene Reelin were co-upregulated in human esophageal carcinoma. AB - Calcium/calmodulin-dependent serine protein kinase (CASK) showed overexpression in human esophageal carcinoma by suppression subtractive hybridization. The upregulation of CASK gene and its target gene Reelin in human esophageal carcinoma tissues versus corresponding normal tissues was revealed by reverse transcription polymerase chain reaction (RT-PCR), immunohistochemistry or Western blot. Moreover, RT-PCR results indicated that the expression patterns of CASK and Reelin in human gastric carcinoma and colon carcinoma were different with those in esophageal carcinoma. Therefore, it suggested that CASK and Reelin were associated with tumorigenesis of esophagus and they were co-upregulated in human esophageal carcinoma. PMID- 11880185 TI - Ezrin, a membrane-cytoskeletal linking protein, is highly expressed in atypical endometrial hyperplasia and uterine endometrioid adenocarcinoma. AB - We previously demonstrated that ezrin transcription was required for in vitro invasion and was involved in the acquisition of metastatic potential in endometrial cancer cells. In order to determine the functional role of ezrin in endometrial cancer, we examined ezrin protein expression in 20 cancerous and 33 non-cancerous tissues by immunohistochemistry and Western blot analysis. The specimens included 20 uterine endometrioid adenocarcinomas (UEC), seven simple endometrial hyperplasias (sH), seven complex endometrial hyperplasias (cH), seven atypical endometrial hyperplasias (aH), and 12 samples of normal endometrium (NE). Tissues of primary (P) and metastatic (M) lesions of endometrial cancers were obtained from five patients. Ezrin was specifically expressed in UEC and its precursor lesions. Ezrin expression was significantly higher in aH (P<0.05) and UEC (P<0.001) compared with NE, sH, and cH. In addition, ezrin was significantly highly expressed in M compared with P (P<0.05). Ezrin expression was associated with neither clinical stage nor histopathologic grade of UEC. In immunohistochemistry, ezrin was localized in the membrane of metastasized cancer cells, although ezrin was mainly distributed in the cytoplasm of most cancer cells and some endometrial hyperplastic cells. On Western blot analysis, ezrin was also detected in both cytosolic and membrane fractions in aH and UEC, whereas ezrin was detected in only cytosolic fraction in sH and cH. These data suggest that expression and subcellular distribution of ezrin protein play an important role in development and progression of UEC. PMID- 11880186 TI - Peroxynitrite-mediated stress is associated with proliferation of human metastatic colorectal carcinoma in the liver. AB - 3-Nitrotyrosine (3-NT), a product of peroxynitrite reaction, is abundantly observed in hepatocytes adjacent to human metastatic colorectal carcinoma. To elucidate its biological significance, we undertook to identify nitric oxide (NO) producing cells and apoptosis under oxidative stress. We observed strong inducible NO-synthase (iNOS) immunoreactivity in the hepatocytes adjacent to metastatic tumor, revealing an identical pattern to 3-NT immunostaining. Furthermore, intense 3-NT immunostaining of hepatocytes was associated with apoptosis whereas carcinoma cells near those hepatocytes presented high proliferating-cell nuclear antigen. Our results suggest that contact of metastatic tumor induces apoptosis in adjacent hepatocytes through peroxynitrite, thus permitting the proliferation of cancer cells. PMID- 11880187 TI - Tumor growth enhancing effects of vascular endothelial growth factor are associated with increased nitric oxide synthase activity and inhibition of apoptosis in human breast carcinoma xenografts. AB - Previously, we demonstrated the significance of vascular endothelial growth factor (VEGF) in promoting the growth of tetracycline-regulated human VEGF165 retroviral vector transduced T47-D breast carcinoma cells, particularly at the early stages of tumor development (Cancer Res. 57 (1997) 3924). Here, we showed histologically that the VEGF overexpressing (VEGF (+)) T47-D cells formed a distinct tumor nodule at day 11, while control cells showed no evidence of replication. The VEGF (+) tumors contained large avascular cavities at days 11 and 21, which were replaced by basement membrane-lined channels at day 30. The number of proliferating tumor cells was not significantly different between the VEGF (+) and control tumors, but the number of apoptotic cells was significantly decreased in the VEGF (+) tumors. Increased nitric oxide synthase (NOS) activity was also observed in the VEGF (+) tumors. These findings indicate that VEGF contributes to tumor growth through inhibition of apoptosis and increased NOS activity, which may be critical during pre-vascular stages of tumor development. PMID- 11880188 TI - Factor(s) released from irradiated B-CLL cells induce apoptosis in leukemic lymphocytes. AB - Photon irradiation of peripheral blood lymphocytes from 25 patients with untreated B-chronic lymphocytic leukemia (B-CLL) induced an increase in apoptotic response by 270%. No significant increase in apoptosis was observed after irradiation of peripheral blood mononuclear cells from 15 healthy volunteers. Supernatants (sups) derived from irradiated leukemic cells incubated with non irradiated autologous cells induced a 75% enhancement in number of apoptotic cells, as compared with sups from non-irradiated CLL cells. The level of tumor necrosis factor alpha, a cytokine known to prevent apoptosis, was reduced in the sups of irradiated CLL cells in comparison to that of non-irradiated lymphocytes. The interleukin (IL)-10 level, an IL reported to induce apoptosis, was similar in the sups of irradiated and non-irradiated lymphocytes from B-CLL patients. No change in IL-2 levels was observed. The significance of these findings and the role of factor(s) in the sups of irradiated leukemic lymphocytes as inducers of apoptosis are discussed. PMID- 11880189 TI - Spatial reorganization of cortical motor output maps of stump muscles in human upper-limb amputees. AB - Spatial changes of the motor cortical representation of the biceps brachii muscle (stump muscle) were studied in ten patients with long-standing amputations at the level of the forearm. Motor output maps were established by focal transcranial magnetic stimulation on a skull surface grid overlying the motor cortex. Characteristics of the motor output map were its spatial extension (number of effective stimulation sites), the maximal response amplitude and the center of gravity (COG) of the spatial distribution of response amplitudes. The extension of the stump muscle motor maps was increased (ratio: 1.5+/-0.3 versus 1.0+/-0.3 in control group; P<0.05) and the stump muscle motor responses were much larger (ratio: 2.6+/-0.6 versus 1.0+/-0.5 in the control group; P<0.05). The COG of the stump muscle map was significantly shifted laterally by, on average, 6.0+/-7.7 mm (range, -3.4-21 mm; P<0.05), either reflecting gross changes of local cortical excitability or structural anatomic reorganization. PMID- 11880190 TI - Asymmetrical mismatch negativity in humans as determined by phonetic but not physical difference. AB - A two-tone oddball procedure was employed to examine the effect of a phonemic category on the mismatch negativity (MMN). One of the stimuli was a phoneme prototype of Japanese /e/, and the other, [e/o], which was perceived by Japanese participants as showing deviance from typicality but is nonetheless included in the category /e/. As control stimuli, a pair of pure tones (1940 and 1794 Hz), corresponding to the F2 frequencies of /e/ and [e/o], respectively, was presented within the same oddball procedure. The MMN for deviant [e/o] revealed greater amplitude than that of deviant /e/, although there was no significant difference in amplitude between the pure tones. The results suggest that a phonemic category determines the auditory sensory memory. PMID- 11880191 TI - Aminophylline aggravates long-term morphological and cognitive damages in status epilepticus in immature rats. AB - Here, we investigated whether aminophylline, an adenosine receptor antagonist used usually as a treatment for premature apnea, had synergistic effects on status epilepticus in the developing brain. On postnatal day 14 (P14), four groups of rats intraperitoneally received saline, aminophylline, lithium- pilocarpine (Li-PC), and Li-PC plus aminophylline, respectively. Subsequently, the Morris water maze task was performed at P80. The brains were then analyzed with cresyl violet stain for histological lesions and evaluated for mossy fiber sprouting with the Timm stain. No seizures were elicited in the saline-treated or aminophylline-treated rats. Both the Li-PC-treated and aminophylline plus Li-PC treated rats exhibited seizures and there was no significant difference in mortality between the two groups. More interestingly, as in adulthood (P80), aminophylline aggravated the spatial deficits and histological damages seen in Li PC-treated rats. In summary, this present study suggests that the use of adenosine receptor antagonists, such as aminophylline, exacerbates seizure induced damage in the developing brain. PMID- 11880192 TI - Extracellular serine protease neuropsin (KLK8) modulates neurite outgrowth and fasciculation of mouse hippocampal neurons in culture. AB - A serine protease neuropsin expressed in the hippocampus of adult brain has been implicated in synaptic plasticity. We report here that endogenous neuropsin was localized extracellularly in neuronal cell bodies and their neurites in mouse hippocampal cultures. Furthermore, we found that, in cultured mouse hippocampal neurons, recombinant neuropsin enhanced neurite projection from soma after 14 h of culture and neuronal aggregation with neurite fascicles at 48 h. This suggests that neuropsin is involved in neurite outgrowth and fasciculation during the development of the nervous system. PMID- 11880193 TI - Neural regulation of hepatic blood flow in rats: an in vivo study. AB - The aim of the present study was to elucidate the influence of the hepatic sympathetic and parasympathetic (vagal) nerves on the hepatic blood flow (HBF), both tonically and when stimulated, using urethane-anesthetized rats as an in vivo experimental model. HBF was measured at the surface of the lateral left lobe of the liver using laser Doppler flowmetry and the hydrogen gas clearance method. Denervation of the hepatic sympathetic nerves had no influence on the HBF, while electrical stimulation of the hepatic sympathetic nerves caused the HBF to decrease in a frequency-dependent manner. This decrease was shown to occur via alpha-adrenergic receptors. In contrast, neither denervation nor electrical stimulation of the hepatic vagal nerves elicited significant changes in the HBF. These results demonstrate that the sympathetic and vagal hepatic nerves have little or no tonic influence on the HBF of rats under urethane anesthesia, whereas the HBF decreases in response to activation of the hepatic sympathetic nerves. PMID- 11880194 TI - Herpes simplex virus type 1-mediated transfer of neurotrophin-3 stimulates survival of chicken auditory sensory neurons. AB - Neurotrophin-3 (NT-3) is one of the most potent stimulators for survival of auditory sensory neurons. Viral transfer of neurotrophins into auditory neurons may offer a route to provide a permanent supply of the growth factor and guarantee their long-term survival. Herpes simplex virus type 1 (HSV-1)-based vectors have demonstrated their effectiveness to transfer genes into peripheral sensory neurons. In the present report, we have produced a HSV-1-based amplicon vector expressing NT-3. This vector efficiently infects isolated auditory neurons and stimulates their survival during distinct developmental stages of the inner ear. Therefore, this vector may present a unique entry point to develop therapies preventing or treating hearing impairment caused by the degeneration of auditory neurons. PMID- 11880195 TI - Insulin-induced expression of the activity-regulated cytoskeleton-associated gene (ARC) in human neuroblastoma cells requires p21(ras), mitogen-activated protein kinase/extracellular regulated kinase and src tyrosine kinases but is protein kinase C-independent. AB - We examined the effect of insulin on the expression of the activity-regulated cytoskeleton-associated gene (ARC), an effector immediate early gene with a proposed role in memory formation. In human SH-SY5Y neuroblastoma cells, application of insulin leads to dramatic increase in ARC mRNA and protein levels. Inhibition experiments reveal, that p21(ras), mitogen-activated protein kinase/extracellular regulated kinase and tyrosine kinase (src) activity are required for the insulin-induced ARC expression in SH-SY5Y cells, whereas protein kinase C is not involved in the signal transduction pathway. Our data indicated for the first time a correlation of the insulin-controlled signal cascade and the induction of synaptic plasticity-associated immediate early genes. PMID- 11880196 TI - Immunocytochemical observation of ghrelin-containing neurons in the rat arcuate nucleus. AB - Ghrelin is a novel peptide that stimulates the release of growth hormone from the pituitary and is involved in hypothalamic feeding regulation. A pre-embedding immunostaining technique was used to study the ultrastructure and synaptic relationships of ghrelin-containing neurons in the rat arcuate nucleus (ARC). Ghrelin-like immunoreactive (ghrelin-LI) neurons were found in the ARC, and were especially abundant in its ventral part. At the electron microscopic level, ghrelin-LI neurons received afferent synapses from many unknown axon terminals. Ghrelin-LI products in the immunoreactive cell bodies, processes, and axon terminals were detected mainly in dense granular vesicles about 110 nm in diameter. Ghrelin-LI presynaptic axon terminals often made synapses with unknown immunonegative neurons. These results suggest that ghrelin acts to regulate food intake through synaptic connections in hypothalamic neuronal networks. PMID- 11880197 TI - Brain oxytocin augments stress-induced long-lasting plasma adrenocorticotropic hormone elevation in rats. AB - Single stress induces long-lasting changes in the hypothalamo-pituitary--adrenal (HPA) axis of adult animals. Selective oxytocin (OXT) receptor antagonist was administrated into the cerebroventricle of male rats to test its influence on plasma adrenocorticotropic hormone (ACTH) responses induced by immobilization stress. The ACTH level is significantly higher than the control level (P<0.05) up to 6 days after single stress. Although the OXT antagonist did not change the plasma ACTH level at the end of single stress (P=0.59), the antagonist significantly decreased the ACTH concentration at the end of repeated (3 days) stress and 2 days after single stress (P<0.05) compared with controls. The results suggest that endogenous brain OXT enhances the long-lasting but not immediate HPA axis response to stress. PMID- 11880198 TI - Analysis of association between the Gln192Arg polymorphism of the paraoxonase gene and schizophrenia in humans. AB - An increasing amount of evidence suggests a possible implication of oxidative stress in the pathophysiology of schizophrenia. Oxidized low-density lipoproteins (oxLDL) have been reported to be capable of eliciting neurocytotoxicity. On the other hand, paraoxonase (PON1), an arylesterase, plays a role in protection against oxidative modifications of LDL and is considered to be one of the antioxidant enzymes. Thus, we investigated the genetic association between a functional polymorphism (Gln192Arg) of the human PON1 gene and schizophrenia in 244 patients and 177 controls. No significant association between the polymorphism and schizophrenia was observed. In addition, our results revealed that there was no association between the genotypes of the polymorphism and any demographic characteristics of patients such as gender, age, age at onset, or current neuroleptic dosage. Our results suggest that the Gln192Arg polymorphism of the PON1 gene may not be involved in the susceptibility to schizophrenia. PMID- 11880199 TI - Human brain cytosolic histamine-N-methyltransferase is decreased in Down syndrome and increased in Pick's disease. AB - Histamine-N-methyltransferase (HMT) inactivates the neurotransmitter histamine. Central histaminergic deficits may contribute to the cognitive impairment of neurodegenerative disorders including Alzheimer's disease (AD) and Down syndrome (DS). However, there is no evidence for histaminergic deficits in Pick's disease (PiD). HMT levels were measured in the frontal cortex and cerebellum of brains of patients with AD, DS, and PiD, and normal aged subjects using proteomics techniques. In frontal cortex, HMT was significantly decreased in DS, but significantly increased in PiD compared with controls. HMT levels were comparable in cerebellum of all groups. Elevated HMT in PiD could lead to increased histamine degradation that in turn would be in agreement with impaired cognitive functions of PiD. Decreased HMT in DS would be compatible with findings of decreased histamine synthesis, thus reflecting a compensation mechanism to antagonize reduced synthesis by decreased degradation. PMID- 11880200 TI - Magnetic responses of human visual cortex to illusory contours. AB - To examine the neural mechanism underlying illusory-contour perception, we measured the magnetic responses of the human visual cortex to an abutting-line grating inducing illusory contours (test stimulus) and a non-abutting-line grating (control stimulus) using the technique of magnetoencephalography (MEG). In the initial latency period of 60-80 ms, the MEG response to the test stimulus was nearly identical with that to the control stimulus, but in the subsequent period of 80-150 ms, the former was larger than the latter. The origin of the peak MEG response to the test stimulus was estimated to be in the vicinity of striate cortex/extrastriate visual cortex for two of the four subjects. These results suggest that, in accord with those of the previous electrophysiological and functional magnetic resonance imaging studies, illusory-contour signals are generated in the very early stage(s) of processing in the primate visual cortex. PMID- 11880201 TI - Cloning and expression of the human N-methyl-D-aspartate receptor subunit NR3A. AB - Native N-methyl-D-aspartate (NMDA) receptors are heteromeric assemblies of four or five subunits. The NMDA receptor subunits, NR1, NR2A, NR2B, NR2C, and NR2D have been cloned in several species, including man. The NR3A subunit, which in rodents is predominantly expressed during early development, seems to function by reducing the NMDA receptor response. The human homologue to the rat NR3A, however, had not been cloned. In order to study the functions of the human NR3A (hNR3A), we have cloned and sequenced the hNR3A. It was found to share 88% of the DNA sequence with the rat gene, corresponding to a 93% homology at the amino acid level. Based on available data from human genome databases, we localized the gene to chromosome 9. The transcript could be detected by in situ hybridization in human fetal spinal cord and forebrain. Two splice variants of NR3A have been reported in rat brain, the longer of the two containing a 60 bp insert in the intracellular domain. We were unable to detect this 60 bp insert in fetal or adult human brain, suggesting that only the short variant is expressed in humans. PMID- 11880202 TI - Caspase-3-associated apoptotic cell death in excitotoxic necrosis of the entorhinal cortex following intraperitoneal injection of kainic acid in the rat. AB - The present study is directed to study: (a) bax translocation and cytochrome c release as mediators of the mitochondrial pathway of apoptosis; (b) Fas-L (Fas ligand) expression as an indicator of the possible involvement of the Fas/Fas-L signaling pathway; and (c) active caspase-3 expression as the main executioner of caspase-mediated apoptosis, in rats receiving an intraperitoneal injection of the glutamate analogue kainic acid (KA) at a dose of 9 mg/kg, which is sufficient to produce generalized seizures and excitotoxic cell death in the entorhinal cortex. Sub-fractionation studies of entorhinal cortex homogenates have shown cytochrome c and cytochrome oxidase IV localized in the mitochondrial fraction, and Bax localized in the cytosolic fraction. No modifications in the sub-cellular distribution of cytochrome c and Bax have been observed at 6 h and 24 h in KA treated rats. Morphological studies have shown cytoplasmic shrinkage and nuclear condensation consistent with necrosis in the entorhinal cortex. Many neurons (about 30% of dying cells) are stained with the method of in situ end-labeling of nuclear DNA fragmentation. Yet only about 5% of dying cells have apoptotic morphology. A percentage of dying cells (5% at 6 h and 40% at 24 h) over-express Fas-L but only about 2% of dying cells at 24 h post-injection express cleaved caspase-3 (17 kD). The present data further support the concept that necrosis is the predominant form of cell death in the entorhinal cortex, although caspase-3 dependent apoptotic cell death may play a limited role, in the present paradigm of KA-induced excitotoxicity. PMID- 11880204 TI - Chemoprevention of human skin cancer. AB - The incidence of skin cancer has been rising in recent years with significant effects on public health. Primary prevention has proven inadequate in impacting the incidence of skin cancer, thus stimulating the development of chemopreventive strategies. The majority of skin cancer chemoprevention studies focus on occurrence of new nonmelanoma skin cancers (NMSC) in individuals with a previous NMSC, or on reduction in the number of premalignant skin lesions such as actinic keratoses (AK). Dysplastic nevi, a likely precursor of melanoma, are also potential targets for chemoprevention strategies. Premalignant lesions are especially attractive as endpoints since they are more common than frank cancer, resulting in reduced sample size, length, and cost of clinical trials. Development of new agents that affect the pathogenesis of skin cancer will be discussed, from elucidation of molecular targets to implementation of trials designed to determine the effects of chemopreventive interventions on human skin cancer. PMID- 11880203 TI - Calpain activation and alpha-spectrin cleavage in rat brain by ethanol. AB - Calpain, a calcium-activated cysteine protease, has been implicated in neuronal degeneration and death. In this study, we have characterized calpain activation in adult rat cerebral cortex and cerebellum, using an experimental paradigm of in vivo chronic ethanol exposure. Ethanol treatment increased the calpain activity in cortex and cerebellum, but to a higher extent in the cortex. Western blot analysis revealed a significant decrease in m-calpain levels while calpastatin levels were unaltered. Calpain activation was further monitored by the proteolysis of alpha-spectrin (fodrin) and protein kinase C-alpha (PKC-alpha). Protease specific spectrin breakdown products revealed calpain generated 150- and 145-kDa fragments. In addition, we also observed a 120-kDa fragment characteristic of caspase-3 activation in the cerebellum. PKC-alpha levels were decreased in the cortex and cerebellum by ethanol. Calpain activation, cleavage of alpha-spectrin into calpain specific signature fragments and decreased PKC alpha protein levels after ethanol treatment provide the evidence of calpain involvement besides caspase-3-mediated cell death in the cortex and cerebellum. Given the role of calpains in cell death, increased calpain activity followed by alpha-spectrin cleavage in this study suggests that calpains are important effectors in ethanol-mediated cell injury and alcoholic neurodegeneration. PMID- 11880205 TI - Natural killer T (NKT) cells and their role in antitumor immunity. AB - Natural killer T (NKT) cells have become a major focus for those who study the innate immune response to tumors and infectious diseases, as well as autoimmunity. These novel T lymphocytes produce both Th1 and Th2 cytokines, recognize phospholipid and glycolipid antigens presented by CD1 molecules in a similar manner as peptides are recognized by cytotoxic T lymphocytes (CTL), and kill tumor cell targets by a perforin-dependent mechanism like NK cells and CTL. These ascribed functions thus demonstrate that NKT cells are a unique cytotoxic effector cell subpopulation with a kaleidoscope of activities. Because they can mediate antitumor effects in vivo with or without the collaboration of NK cells, the study of NKT cells in antitumor immunity may lead to novel treatments based on the ability to manipulate the generation and/or activity of these multifunctional lymphocytes. PMID- 11880207 TI - Radiotherapy-related fatigue. AB - Radiotherapy-induced fatigue is a common early and chronic side-effect of irradiation, reported in up to 80 and 30% of patients during radiation therapy and at follow-up visits, respectively. It is frequently underestimated by medical and nursing staff, only about 50% of patients discuss it with a physician and in one fourth of cases any intervention is proposed to the patient. The patients rarely expect fatigue to be a side-effect of treatment. The etiology of this common symptom, its correlates and prevalence are poorly understood. In numerous studies the level and time course of fatigue was demonstrated to depend on the site of tumor and treatment modalities. For example, psychological mechanisms have been proposed to explain fatigue in women receiving irradiation for early breast cancer, whereas decline in neuromuscular efficiency rather than psychological reasons can lead to the fatigue observed in patients undergoing radiotherapy for prostate cancer. Fatigue can affect global quality of life more than pain, sexual dysfunction and other cancer- or treatment-related symptoms. Several interventions have been tested in the management of radiotherapy-related fatigue and some randomized studies have been recently published. Although an optimal method has not yet been established, some promising results have been reported with relaxation therapy, group psychotherapy, physical exercise and sleep. Further methodologically correct studies are warranted to define better the causes, optimal prevention and management of this symptom. PMID- 11880206 TI - AIDS-related tumors: integrating antiviral and anticancer therapy. AB - The introduction of highly active antiretroviral therapy (HAART) has changed dramatically the landscape of HIV disease. Deaths from AIDS-related diseases have been reduced by 75% since protease inhibitor therapy and combination antiretroviral therapy came into use in late 1995. While KS is declining, the situation for non-Hodgkin's lymphoma is more complex with a reduced incidence of primary central nervous system lymphoma, but a relative stability in the number of patients developing systemic NHL. AIDS-related NHL appears not to be markedly decreased by the introduction of HAART and it is the greatest therapeutic challenge in the area of AIDS oncology. The emphasis has now shifted to cure while maintaining vigilance regarding the unique vulnerability of HIV-infected hosts. Furthermore, also for the prolongation of the survival expectancy of these patients, other non-AIDS-defining tumors, such as Hodgkin's disease, anal, head and neck, lung and testicular cancer, and melanoma have been recently reported with increased frequency in patients with HIV infection. PMID- 11880209 TI - Secondary cytoreductive surgery in the patient with recurrent ovarian cancer is often beneficial. AB - The majority of women with ovarian cancer will present at an advanced stage and ultimately experience a recrudescence of tumor. Recent data indicates that secondary cytoreductive surgery is feasible, well tolerated and associated with significant prolongation of survival in selected patients with recurrent ovarian cancer. PMID- 11880208 TI - Prognostic and predictive factors of immunotherapy in metastatic renal cell carcinoma. AB - Metastatic renal cell carcinoma has a poor prognosis. The value of immunotherapy with IFN-alpha and IL-2 both as single agent or as the combination is extensively investigated. The optimal dose and schedule is not known. In various studies response rates vary between 10 and 40%. The duration of response is variable. For a partial response a median duration between 10 and 12 months is given. Complete responses are sometimes long-lasting (a couple of years). The toxicity is drug, dose and schedule dependent. On the basis of a number of prognostic factors, such as performance score, time between the initial diagnosis and the treatment of metastases and the number of metastatic sites, patients can be divided in different prognostic groups. Patients who are classified in the good or intermediate prognostic group may have an improvement of their survival after immunotherapy and therefore they are candidates for immunotherapy. PMID- 11880210 TI - Geriatric medical oncology in the care of elderly cancer patients. AB - Most cancers are diagnosed after 70 years of age but standard management and treatment for elderly cancer patients remain to be established. To determine whether the availability and recognition of medical oncology may influence cancer care in this population, five successive periods were studied. The number of formal written consultations given at the geriatric hospital and at the center for continuous care was measured. Over a period of 36 months, the number of consultations rose from an initial 1.5% (n=26) and 3.8% (n=25) to 3% (n=71) and 12.5% (n=103) of the respective total number of admissions. As expected, the increase exactly matched both main geographical location and functional position of the single appointed medical oncologist. However, following the definitive establishment of the medical oncologist at the center for continuous care, the number of formal written consultations markedly decreased. At the geriatric hospital, medical oncology returned to the 'status quo ante' whereas it was quite simply incorporated in the daily care of all cancer patients at the center for continuous care. Thus, improvement of cancer care in the elderly may first depend on human resources such as trained specialists to make a true difference with the prior situation. PMID- 11880211 TI - Chemotherapy of lung cancer in the elderly. AB - In the last few years it has become evident that the elderly lung cancer patient represents a peculiar individual with regard to both the tolerance to the tumor and its treatment. Age per se cannot be considered an adverse prognostic factor, however, the physiologic impairment of the functions of important organs like liver, kidney, bone marrow etc., may render more unpredictable the treatment related toxicity, and moreover, the higher incidence of concomitant diseases which occurs with aging certainly translates in a worse survival outcome. As a consequence, a careful multidimensional evaluation (functional, emotional, socioeconomic status, comorbidities, etc.) should be preliminarily performed in patients eligible for chemotherapy treatment. Several approaches have been tested in elderly lung cancer patients. Different therapeutic attitudes exist, which first take into account the kind of histology (small cell lung cancer, SCLC and non-small cell lung cancer, NSCLC). Of course a more aggressive approach can be sometimes justified in an elderly SCLC patient in view of the high responsiveness of this disease, while more concerns exist about the use of aggressive chemotherapy regimen in elderly patients with NSCLC histology. In view of these considerations, more clinical trials are being planned to specifically assess the role of chemotherapy in this subset of patients.A brief review of the most important phase II and III trials conducted in elderly patients with either SCLC or NSCLC is provided here. A description of the most important still unsolved issues will be made, and an outline of the ongoing clinical trials in these patients will be provided. PMID- 11880212 TI - Roles and responsibilities of epidemiologists. AB - Two distinct views of the roles and responsibilities of epidemiologists have emerged in a decades-long debate: one keeps professional practice constrained to science; the other adds active participation in public health policymaking. In defense of the narrower view are several claims: that epidemiologists lack expertise in policymaking; that advocating policy adversely affects scientific objectivity; that the limits of epidemiologic science work against translating results into policy; and that participation in policy can bring on personal attacks. In this study, each claim is addressed. Epidemiologists already participate fully in educational, research funding, and editorial policymaking and thereby have an experiential foundation in some of the basics of policymaking. Policymaking can enhance scientific objectivity because it requires not only the use but more importantly the improvement of empirical methods. Finally, the comforts of professional life are not the primary yardsticks of our responsibilities. Arguments in favor of active involvement in public health policymaking are presented. Epidemiologists have been mixing science and policymaking for a long time and there is a strong sense that the benefits of public stewardship outweigh the risks. The American College of Epidemiology's Ethics Guidelines support this view. Active participation in public heath policymaking will, however, require curriculum changes in graduate training programs. With additional training and a broader recognition that public health policymaking is an appropriate professional pursuit, epidemiologists can look to a bright and challenging future in the science and practice of public health. PMID- 11880213 TI - Considerations on guidelines of epidemiologic practice. AB - Epidemiologists in the United States have been intrigued for almost two decades by the prospect of guidelines of epidemiologic practice: an issue partially raised by a perceived need for criteria that may distinguish advocacy positions from more conservative interpretations in epidemiology. Preliminary efforts have considered questions of subject protection, of study structure and documentation, and of professional etiquette. Deeper ethical interests have been coalescing around the need for candor and truth in the interpretation of uncertain epidemiologic findings--a need construed to be central to trustworthiness, and whose consideration would bring about a reappraisal of other aspects of epidemiologic practice. Openness about uncertainties would likely develop naturally into a search for ways to measure the relative strength of epidemiologic warnings, as an aid to ranking public health priorities. Principles aimed at resolving ethical, operational, and priority questions become interdependent elements of professional guidelines for epidemiology. PMID- 11880214 TI - Accuracy of lipoprotein lipids and apoproteins in predicting coronary heart disease in diabetic American Indians. The Strong Heart Study. AB - PURPOSE: To evaluate the accuracy of lipoprotein lipids and apoproteins in predicting coronary heart disease (CHD) in diabetic American Indians. METHODS: This study included 2099 diabetic participants of the Strong Heart Study, which is a longitudinal study of cardiovascular disease (CVD) and its risk factors in American Indians in Arizona, Oklahoma, and South and North Dakota. Diabetic participants with incident CHD (N = 126) were selected as the case group, and those without CHD or any cardiovascular events were the control group (N = 1732). Previous vascular events such as stroke were the sole exclusion criterion (N = 241). Baseline measurements of lipoprotein lipids and apoproteins were used to predict CHD diagnosed at the 4-year follow-up examination by using Receiver Operating Characteristic (ROC) curve analysis. RESULTS: The ratio of high-density lipoprotein (HDL) to total cholesterol had the highest area under the ROC curve (0.69 +/- 0.02). The areas for the ratios of HDL to low-density lipoprotein (LDL) cholesterol (0.68 +/- 0.02), apo AI/B (0.66 +/- 0.02), and the single component of total cholesterol (0.64 +/- 0.03) and LDL cholesterol (0.63 +/- 0.05) were not significantly different from the area for HDL/total cholesterol. However, the areas for apo B (0.64 +/- 0.02), HDL cholesterol (0.62 +/- 0.03), triglycerides (0.58 +/- 0.03), and apo AI (0.57 +/- 0.05) were significantly lower than the area for HDL/total cholesterol. Logistic regression analysis indicates that only HDL and LDL cholesterol were significant independent lipoprotein lipid and apoprotein predictors for CHD. The other significant predictors in the model were study center, age, gender, and albuminuria. The ROC area for this model is 0.75. CONCLUSIONS: HDL and LDL cholesterol were the most important independent predictors for incident CHD in diabetic American Indians. The ratios of HDL/total cholesterol, HDL/LDL cholesterol, and apo AI/B had higher accuracy for predicting CHD. Although the values for all lipoprotein lipids and apoproteins and their ratios were not large enough to definitely predict CHD, they can be used as screening tools for CHD in diabetic American Indians. PMID- 11880215 TI - The effect of physicians' training on prescribing beta-blockers for secondary prevention of myocardial infarction in the elderly. AB - PURPOSE: The objective was to determine whether the year and medical school of graduation, the medical specialty and the sex of the treating physician was associated with prescribing beta-blockers after hospital discharge among survivors of myocardial infarction (MI), after adjusting for patient characteristics. METHODS: We carried out a dynamic retrospective cohort study using data from a longitudinal database that contained information on Quebec residents over the age of 65 years sent home from hospital after MI between 1990 and 1993. The outcome was a beta-blocker being dispensed after hospital discharge. Logistic regression was used to estimate the association between training characteristics and beta-blocker dispensation and clustering of patients within physicians was accounted for using Generalized Estimating Equations. RESULTS: The cohort consisted of 14,334 MI survivors who were treated by 3209 physicians, yielding a mean of about 4.5 patients per physician [standard deviation (SD) = 8.2]. Beta-blockers were prescribed to approximately one-third of subjects. After adjusting for patients' demographic characteristics, comorbid medical conditions, and markers of MI severity, physicians who were more likely to prescribe a beta-blocker included cardiologists and the most recent graduates (graduating after 1989). Systematic differences were also observed between graduates of different medical schools. CONCLUSIONS: After adjusting for differences between patients', the sex of the physician was largely unrelated to prescribing beta-blockers for secondary prevention of MI. However, prescribing differed by training characteristics such as medical specialty and year and medical school of graduation. PMID- 11880216 TI - Gamma-glutamyltransferase, alcohol, and blood pressure. A four year follow-up study. AB - PURPOSE: We performed this study to see: 1) whether gamma-glutamyltransferase (GGT) was a predictor for hypertension irrespective of amount of alcohol consumption; and 2) whether the relationship between alcohol consumption and blood pressure (BP) was different depending on GGT. METHODS: This study included 8170 healthy male workers in a steel manufacturing company who had undergone health examinations in both 1994 and 1998. RESULTS: The adjusted relative risk (RR) for hypertension over four years among those with GGT >or=30 U/L at baseline was 1.6 (95% confidence interval (CI): 1.1-2.4), compared to those with GGT < 30 U/L. This relationship was shown only in drinkers. Among subjects with GGT < 30U/L, no matter how much the subjects drank, the risk of hypertension in drinkers was similar with that of non-drinkers. However, among those with GGT >or=30 U/L, adjusted RRs for light, moderate, and heavy drinkers compared to non drinkers were 1.4 (95% CI: 0.5-4.5), 5.2 (95% CI: 1.5-18.0), and 5.3 (95% CI: 1.0 27.6). CONCLUSIONS: This study showed that elevated GGT could be a predictor for hypertension in drinkers and the relationship between alcohol consumption and hypertension was shown only among those with GGT >or= 30 U/L at baseline. These findings suggest that increased serum GGT levels may reflect individual susceptibility to the blood pressure raising effect of alcohol. PMID- 11880218 TI - Lipoprotein(a) in American Indians is low and not independently associated with cardiovascular disease. The Strong Heart Study. AB - PURPOSE: To evaluate the distribution of lipoprotein(a) (Lp(a)) and assess its association to cardiovascular disease (CVD) in American Indians. METHODS: Lp(a) was measured in 3991 American Indians (aged 45-74 years with no prior history of CVD at baseline) from 13 communities in Arizona, Oklahoma, and South/North Dakota. They were followed prospectively from 1989 to 1997 for CVD. The distribution of Lp(a) was examined by center, sex, and diabetic status. Spearman correlation coefficients and Cox regression models were used to evaluate the association of Lp(a) to CVD. RESULTS: A total of 388 participants subsequently developed CVD. Median Lp(a) concentration in American Indians was 3.0 mg/dl. This was almost half of that in whites and one sixth in blacks from the CARDIA study measured by the same method. Nondiabetic participants had significantly higher Lp(a) levels than diabetic participants for both genders. Lp(a) levels were higher in women than in men for nondiabetic participants, but there was no gender difference for diabetic participants. Correlation analysis showed Lp(a) was significantly negatively correlated with the degree of Indian heritage, insulin, triglycerides (TG), fasting plasma glucose (FPG), and 2-hour plasma glucose (2hPG), and positively with low-density lipoproteins (LDL), apoprotein B (apoB), and fibrinogen (FIB). In Cox regression models, adjusting for other risk factors, Lp(a) was no longer a significant predictor of CVD in either diabetic or nondiabetic participants. CONCLUSIONS: The lower concentration of Lp(a) in American Indians and the high correlation with Indian heritage confirm the concept that Lp(a) concentration is in large part genetically determined. Lp(a) concentration is not an independent predictor of CVD among American Indians; it is higher in those who develop CVD because of its positive correlation with LDL, apoB, and FIB. PMID- 11880217 TI - Changes in cardiovascular disease risk factors among American Indians. The Strong Heart Study. AB - PURPOSE: This study describes changes in cardiovascular disease (CVD) risk factors in older American Indians over a 4-year period. METHODS: The Strong Heart Study, a longitudinal population-based study of CVD and CVD risk factors among American Indians aged 45-74 years, measured CVD risk factors among 3638 members of 13 tribes in three geographic areas during examinations in 1989 to 1991 and 1993 to 1995. RESULTS: Changes in mean low-density lipoprotein (LDL) cholesterol and the prevalence of elevated LDL cholesterol were inconsistent. Mean high- density lipoprotein (HDL) cholesterol decreased, and the prevalence of low HDL cholesterol increased throughout. Mean systolic blood pressure and hypertension rates increased in nearly all center-sex groups, and hypertension awareness and treatment improved. Smoking rates decreased but remained higher than national rates except among Arizona women. Mean weight and percentage body fat decreased in nearly all center-sex groups but the prevalence of obesity did not change significantly in any group. Diabetes and albuminuria prevalence rates increased throughout the study population. The prevalence of alcohol use decreased, but binge drinking remained common in those who continued to drink. CONCLUSIONS: Improvements in management and prevention of hypertension, diabetes, renal disease, and obesity, and programs to further reduce smoking and alcohol abuse, are urgently needed. PMID- 11880219 TI - Hospitalized congestive heart failure after renal transplantation in the United States. AB - PURPOSE: African Americans have increased risk for congestive heart failure (CHF) compared to Caucasians in the general population, but the risk of CHF in African American renal transplant recipients has not been studied in a national renal transplant population. METHODS: Therefore, 33,479 renal transplant recipients in the United States Renal Data System (USRDS) from 1 July, 1994 to 30 June, 1997 were analyzed in an historical cohort study of the incidence, associated factors, and mortality of hospitalizations with a primary discharge diagnosis of CHF [International Classification of Diseases-9 (ICD9) Code 428.x]. RESULTS: African American renal transplant recipients had increased age-adjusted risk of hospitalizations for congestive heart failure compared to African Americans in the general population [rate ratio 4.60, 95% confidence interval (CI) 4.59-4.62]. In logistic regression analysis, African American recipients had increased risk of congestive heart failure after renal transplantation, independent of other factors. Among other significant factors associated with congestive heart failure, the strongest were graft loss and allograft rejection. No maintenance immunosuppressive medications were associated with CHF. In Cox regression analysis patients hospitalized for CHF had increased all-cause mortality compared with all other recipients (hazard ratio 3.69, 95% CI, 2.23-6.10), but African American recipients with CHF were not at significantly increased risk of mortality compared to Caucasian recipients with CHF. CONCLUSIONS: African Americans recipients were at high risk for CHF after transplant independent of other factors. The reasons for this increased risk should be the subject of further study. All potential transplant recipients should receive particular attention for the diagnosis and prevention of CHF in the transplant evaluation process, which includes preservation of allograft function. PMID- 11880220 TI - Cumulative abuse experiences, physical health and health behaviors. AB - PURPOSE: Our purpose was to investigate the complex relationship between a range of lifetime abuse experiences with current physical health and health behaviors. METHODS: Between October 1998 and May 1999, interviews were conducted with 557 ethnically diverse women seen at two urban primary care centers. Seven forms of abuse were measured: childhood physical and sexual abuse, past physical and sexual intimate partner violence (IPV), and recent emotional, physical, and sexual IPV. Severity was measured for six of these forms. Multiple non-specific physical symptoms were measured with a modified PRIME-MD, and four health behaviors were ascertained. RESULTS: Approximately 10% of women never experiencing abuse reported multiple non-specific physical symptoms, compared with 25.8% to 78.4% of women reporting a range of abuse experiences. Increases in recent IPV, past IPV, child abuse, and economic hardship were associated with increases in reported symptoms. Women who experienced IPV were more likely to report smoking cigarettes, binge drinking, and having poor nutritional habits. CONCLUSIONS: Recent IPV is associated with physical symptoms and risky health behaviors beyond the effects of child abuse, past IPV, and economic disadvantage. Understanding a person's IPV experiences may inform interventions for health behaviors, such as smoking cessation programs. PMID- 11880221 TI - Hyperinsulinemia, dyslipidemia, and obesity as risk factors for hospitalized gallbladder disease. A prospective study. AB - PURPOSE: Prospective studies of gallbladder disease have investigated a limited number of risk factors and have been conducted predominantly in women. Determinants of hospitalized gallbladder disease were examined in a large, population-based cohort of men and women. METHODS: Subjects, aged 45-64 years, were participants in the Atherosclerosis Risk in Communities (ARIC) Study who reported no history of gallbladder disease at baseline (n = 12,773). Incident cases of gallbladder disease were identified through surveillance of hospital discharges between the baseline visit (1987-1989) and 1996. RESULTS: The crude incidence rate of hospitalized gallbladder disease was 3.8 per 1000 person-years. In women, increasing risk was observed for increasing levels of body-mass index and waist-to-hip ratio, whereas in men, increased risk was observed only in the morbidly obese (BMI >or= 35). Fasting serum insulin, low HDL cholesterol, elevated triglycerides, and hormone replacement therapy were all positively associated with gallbladder disease risk. The relative risks associated with having one, two, or three or more components of the "multiple metabolic syndrome" in men were 1.45 (95% CI = 0.9-2.3), 2.17 (1.3-3.6), and 2.34 (1.3-4.3), respectively. CONCLUSIONS: In men, hyperinsulinemia and dyslipidemia may have some role in the etiology of gallbladder disease beyond their association with obesity, whereas in women, increased body size, central adiposity, and hormone replacement therapy may be more important determinants of gallbladder disease. PMID- 11880222 TI - Recombinant Mycobacterium bovis BCG producing the circumsporozoite protein of Plasmodium falciparum FCC-1/HN strain induces strong immune responses in BALB/c mice. AB - The current vaccine against tuberculosis, Mycobacterium bovis strain bacillus Calmette-Guerin (BCG), offers potential advantages as a live, innately immunogenic vaccine vehicle for expression and delivery of protective recombinant antigens. Malaria is one of the severest parasitic diseases in humans especially in the developing world. No efficacious vaccine is currently available. However, circumsporozoite protein (CSP) is a malaria vaccine candidate currently undergoing clinical trials. We analyzed the immune response to recombinant BCG (rBCG) vaccine expressing Plasmodium falciparum CSP (BCG-CSP) under the control of heat shock protein 70 promoter in BALB/c mice. The lymphocytes proliferative response to P. falciparum soluble antigen was significantly higher than those in the groups of BCG and normal saline, and the production of cytokines (IFN-gamma and IL-2) in response to malaria antigen was significantly higher in rBCG and BCG groups than control group of normal saline. A specific IgG antibody response against P. falciparum antigen of CSP was also characterized. The booster injection could enhance the production of cytokine, proliferation responses of spleen lymphocytes and the antibodies titer of BCG-CSP. The results in the study demonstrated that rBCG vaccine producing CSP is an appropriate vaccine for further evaluation in non-human primates. PMID- 11880224 TI - A rodent malaria, Plasmodium berghei, is experimentally transmitted to mice by merely probing of infective mosquito, Anopheles stephensi. AB - We found that infection of a rodent malaria, Plasmodium berghei, occurred when the sporozoites were injected into the skin, the muscle, the peritoneal cavity and the tail end. Mice, which were injected with sporozoites in the tail end and had the site cut 5 min later, did not develop malaria. We also found that mice developed malaria when malaria infective mosquitoes, Anopheles stephensi, were forced not to take blood but only to probe into the skin. Moreover, the mice probed by the infective mosquitoes were protected from malaria infection if the site was treated with Kyu (heat treatment) after the mosquitoes had probed. These findings indicate that malaria infection occurs not only by blood feeding of the infective mosquito but also by probing of the mosquito. Sporozoites injected into the skin remain at the injected site for at least 5 min, then migrate to the blood vessels and invade into the blood stream. At present, the mechanism is not clear, although we propose here the existence of the skin stage of malaria parasites before the liver stage and the blood stage. PMID- 11880223 TI - Mitochondrial-type hsp70 genes of the amitochondriate protists, Giardia intestinalis, Entamoeba histolytica and two microsporidians. AB - Genes encoding putative mitochondrial-type heat shock protein 70 (mit-hsp70) were isolated and sequenced from amitochondriate protists, Giardia intestinalis, Entamoeba histolytica, and two microsporidians, Encephalitozoon hellem and Glugea plecoglossi. The deduced mit-hsp70 sequences were analyzed by sequence alignments and phylogenetic reconstructions. The mit-hsp70 sequence of these four amitochondriate protists were divergent from other mit-hsp70 sequences of mitochondriate eukaryotes. However, all of these sequences were clearly located within a eukaryotic mitochondrial clade in the tree including various type hsp70 sequences, supporting the emerging notion that none of these amitochondriate lineages are primitively amitochodrial, but lost their mitochondria secondarily in their evolutionary past. PMID- 11880225 TI - Biochemical and enzymatic characterization of a partially purified casein kinase 1 like activity from Trypanosoma cruzi. AB - Two protein kinase activities that use casein as a substrate, Q-I and Q-II, were identified in the epimastigote stage of Trypanosoma cruzi upon chromatography on Q-Sepharose. Q-I was purified further through concanavalin A-sepharose (Q-I*) to remove any trace of the contaminating protease cruzipain. The optimal activity for Q-I* was obtained at pH 8.0, 25 degreesC, 5 mM MgCl(2) and 75 mM NaCl. The size and pI of Q-I* were determined to be 33-36 kDa and 9.6, respectively. When two selective peptide substrates for casein kinases (CKs) (P1: RRKDLHDDEEDEAMSITA for CK1 and P2: RRRADDSDDDDD for CK2) were used, Q-I* was shown to specifically phosphorylate P1. Kinetic studies showed that Q-I* has a K(m) of 5.3 +/- 0.34 mg/ml for casein, 157.6 +/- 5.3 microM for P1 and 35.9 +/- 3.9 microM for ATP. The enzyme was inhibited by N-(2-amino-ethyl)-5-chloroisoquinoline-8-sulfonamide (CKI-7) or 1-(5-chloroisoquinoline-8-sulfonyl) (CKI-8), two inactivators of mammalian CKs. CKI-7 behaved as a competitive inhibitor with respect to ATP, with a K(I) of 75-100 microM. Treatment with high concentrations of polylysine or heparin also resulted in a significant inhibition of Q-I*. Two well-known activators of mammalian CKs, spermine and spermidine, were also tested. Spermine and spermidine activated Q-I* in a dose-dependent manner. Based on the following characteristics: (1) the ionic strength required for elution from anion-exchange resins; (2) its molecular size and monomeric structure; (3) pI; (4) high level of specificity for P1; (5) inactivation by CKI-7 and CKI-8; and (6) insensitivity to GTP and low concentrations of heparin, we conclude that Q-I* belongs to the CK1 family of protein kinases. PMID- 11880226 TI - Trichomonas vaginalis: characterization of a family of P-type ATPase genes. AB - P-type ATPases are ion-transporting pumps that enable organisms to control cellular functions and survive changing environmental conditions by regulating internal ion concentrations. Eight P-type ATPases were identified in the amitochondriate protist Trichomonas vaginalis using polymerase chain reaction (PCR) amplification with oligonucleotide primers that recognize conserved motifs present in all P-type ATPases, the ATP phosphorylation site (DKTGTLT) and the ATP binding site (TGDGVND). Phylogenetic analysis and the presence of conserved motifs in predicted peptide sequences identify the Trichomonas ATPases as a sarcoplasmic-endoplasmic reticulum calcium pump (TVCA1); three additional Ca(2+) transporting pumps (TVCA2-4), three phospholipid translocases (TVAPLT1-3), and one P-type ATPase of unknown transport specificity (TVPATP8). Southern blot analyses indicate that the P-type ATPase genes are not linked and are present in single copy, except TVCA2 and TVCA4 which contain additional copies or closely related homologues within the genome. Transcripts of 3.1 kb for TVCA1, 3.0 kb for TVCA2, 2.9 kb for TVCA3, 4.0 kb for TVAPLT1, 4.2 kb for TVAPLT2, 3.9 kb for TVAPLT3, and 3.1 kb for TVPATP8 were detected by Northern blot analysis. No TVCA4 transcript was observed, however, RT-PCR amplification of a transcript product indicates that TVCA4 is expressed. RNA expression of the Trichomonas ATPases, except TVCA3, was constitutive over a range of environmental conditions. TVCA1, TVAPLT3 and TVPATP8 had the highest levels of RNA expression while TVAPLT1 and TVAPLT2 expression was the lowest. PMID- 11880227 TI - Response to carbon dioxide by the infective larvae of three species of parasitic nematodes. AB - The response of infective third-stage larvae (L3) of three species of parasitic nematodes, Ancylostoma caninum, Strongyloides stercoralis, and Haemonchus contortus to carbon dioxide (CO(2)) at physiological concentrations was investigated. L3 of the skin-penetrating species, A. caninum and S. stercoralis, were stimulated by CO(2) at the concentration found in human breath (3.3-4%); these larvae responded by crawling actively, but not directionally. Crawling was not stimulated by breath passed through a CO(2)-removing "scrubber" or by "bench air". Both A. caninum and S. stercoralis L3 stopped crawling when exposed to 5% CO(2) for 1 min. L3 of A. caninum became active 9-14 min after exposure to 5% CO(2) ended, but activity resumed more rapidly (10-15 s) if larvae were subsequently exposed to breath or breath through the scrubber. L3 of S. stercoralis resumed crawling 30-35 s after exposure to 5% CO(2), but resumed crawling within a very few seconds when exposed to breath or breath through the scrubber. Thus, while 5% CO(2) was inhibitory, lower concentrations of this gas stimulated L3 of both species. Apparently, exposing immobilized larvae to breath or breath through the scrubber causes the environmental CO(2) concentration to drop to a level that is stimulatory. The L3 of H. contortus ceased crawling and coiled when exposed to human breath or to 1% CO(2), but continued to move within the coil in both cases. The crawling response of the L3 of the two skin penetrating species, A. caninum and S. stercoralis, to stimulation by CO(2) probably relates to their active host-finding behavior, while the cessation response elicited by CO(2) in H. contortus larvae may relate to the fact that they rely on passive ingestion by a ruminant host. PMID- 11880228 TI - The expression system of biologically active canine interleukin-8 in Leishmania promastigotes. AB - It has been reported that Leishmania promastigotes have ability to express foreign genes on drug selectable plasmids. To investigate further abilities of the recently described expression vector, P6.5, in the transfection of Leishmania organisms (Chen D-Q, Kolli BK, Yadava N et al. Episomal expression of specific sense and antisense mRNAs in Leishmania amazonensis: modulation of gp63 levels in promastigotes and their infection of macrophages in vitro. Infect Immun 2000;68:80--86), the constructed expression vector, which contains canine interleukin-8 (cIL-8) coding cDNA, was introduced by electroporation to promastigotes of four species of the genus Leishmania: Leishmania amazonensis, L. equatorensis, L. donovani and L. infantum. Extrachromosomal DNAs and total RNAs from the transfected promastigotes were subjected to polymerase chain reaction (PCR) and reverse transcriptase-PCR, respectively, using cIL-8 gene specific primers, and a predicted product of 330 bp was detected. Western blot analysis using a mouse monoclonal antibody raised against cIL-8 demonstrated the successful expression of cIL-8 in the transfectants and culture supernatants. Culture supernatants of the transfected L. amazonensis and L. equatorensis promastigotes showed a high chemotactic activity to both dog and mouse polymorphonuclear leukocytes. These results indicate that Leishmania promastigotes transfected with the expression vector P6.5 containing cIL-8 cDNA are capable of producing biologically active cIL-8. The Leishmania expression system using the P6.5 vector might be a useful alternative for the production of biologically active recombinant cytokines. PMID- 11880229 TI - A mass occurrence of human infection with Diplogonoporus grandis (Cestoda: Diphyllobothriidae) in Shizuoka Prefecture, central Japan. AB - Forty-six cases of human infection with Diplogonoporus grandis were found in Shizuoka Prefecture on the Pacific coast of Central Japan in 1996. The cases were predominantly elderly male patients over 50 years of age. Although all cases were reported from May to September of the year, most of them were diagnosed in June and July. We suspected that the transmission was due to the consumption of raw juvenile Japanese anchovy (Engraulis japonicus), which are seasonally caught in the spring months off the Pacific coast of the Prefecture. In almost a hundred years after its discovery in 1894, there had been more than 180 cases of human diplogonoporiasis recorded in Japan. The high incidence within a relatively short time frame of our investigation is regarded unusual even in this country. PMID- 11880230 TI - Gastric hyperplasia and parietal cell loss in Taenia taeniaeformis inoculated immunodeficient mice. AB - Immunodeficient mice were studied to determine their suitability as models in investigating the role of Taenia taeniaeformis larval products in the development of gastric hyperplasia. Recombinant active gene 2 (RAG2)-deficient and severe combined immune-deficient (SCID) mice were studied as candidate animal models. RAG2-deficient mice inoculated orally with T. taeniaeformis eggs developed gastric hyperplasia with alcian blue-periodic acid-Schiff-positive cell proliferation similar to those of rats. SCID mice inoculated with different doses and routes of T. taeniaeformis in vitro-hatched oncospheres and those orally inoculated with eggs resulted also in different degrees of gastric hyperplasia. Influence of inoculation forms of parasite, doses and routes of inoculation on initiation of hyperplastic gastropathy was suggested to be dependent on number and size of developed larvae. Both RAG2-deficient and SCID mice with hyperplastic mucosa were observed with significant loss of parietal cells. Apparent decrease in parietal cell number was observed in SCID mice at 2 weeks after intraperitoneal inoculation with oncospheres before hyperplastic lesions developed. Earliest occurrence of gastric hyperplasia in SCID mice was observed at 3 weeks after oral inoculation of in vitro-hatched oncospheres, sooner than orally inoculated rats. The results suggested that these immunodeficient mice could be used as animal models to study factors involved in T. taeniaeformis induced gastric mucous cell hyperplasia. PMID- 11880232 TI - Stage specificity of Plasmodium falciparum telomerase and its inhibition by berberine. AB - Telomerase activity in synchronized Plasmodium falciparum during its erythrocytic cycle was examined using the TRAP assay. Telomerase activity was detected at all stages of the parasite intraerythrocyte development, with higher activity in trophozoite and schizont stages compared with ring form. Berberine, extracted from Arcangelisia flava (L.) Merr., inhibited telomerase activity in a dose dependent manner over a range of 30-300 microM, indicating that P. falciparum telomerase might be a potential target for future malaria chemotherapy. PMID- 11880231 TI - Tyvelose and protective responses to the intestinal stages of Trichinella spiralis. AB - The unusual sugar tyvelose is the immunodominant portion of the major larval glycoprotein antigens of Trichinella spiralis, which play an important role in generating immunity against the intestinal stages of infection. The possibility that the tyvelose component itself may have a host- or parasite-protective role in the intestine was tested by following the outcome of challenge infections in mice primed and boosted with tyvelose-BSA, or in mice primed with tyvelose-BSA before boosting with larval antigen. Although antibody responses were raised against tyvelose there was no evidence of protective immunity against the intestinal stages, as assessed by total adult worm recovery or by size and fecundity of female worms in immunized mice. Equally, priming with tyvelose-BSA before boosting with larval antigen had no effect on the expression of immunity against a challenge infection. The predominant antibody isotype recorded in all immunized mice was IgG1, suggesting the induction of type 2 T cell responses, and this was confirmed by cytokine analysis, mesenteric node lymphocytes of all mice showing production of IL-5 but not IFN-gamma. Clearly immunization with tyvelose had no significant effect on T cell polarization. The data show that, with the experimental design employed, there was no evidence for a functional role of tyvelose in either host- or parasite-protection during the intestinal phase of infection. PMID- 11880233 TI - First outbreak of Baylisascaris procyonis larva migrans in rabbits in Japan. AB - A syndrome of progressive neurological signs was noticed in rabbits (Oryctolagus cuniculus) kept in a small wildlife park in mid-July 2000. Three out of 12 common raccoons (Procyon lotor) kept in this park were infected with Baylisascaris procyonis, and the larvae were found from affected rabbits. This outbreak is the first proven B. procyonis larva migrans in Japan, and the potential risk of serious zoonosis by this ascarid species should be considered by pet owners, veterinarians, physicians and public health authorities in this country as in North America where raccoons are endemic. PMID- 11880234 TI - The susceptibility of inbred mice to infection with Brachylaima cribbi (Digenea: Brachylaimidae). AB - Susceptibility to infection with Brachylaima cribbi was studied in eight strains of inbred mice (AKR, C3H/HeJ, CBA/CaH, BALB/c, DBA/2J, SJL/J, A/J, C57BL/6J) and Swiss albino outbred mice by quantifying faecal egg excretion over the period of the infection. Preliminary experiments indicated that a combination of filtration/sedimentation/diethyl ether sedimentation was the most sensitive and reliable technique for quantification of eggs in faeces. Mice were infected with 13-15 wild-type B. cribbi metacercariae from naturally infected Cernuella virgata and in a second experiment with human-derived B. cribbi from laboratory-reared Helix aspersa. In both experiments C57BL/6J mice were the most susceptible having the highest egg excretion levels and the longest duration of infection. Worm burdens were assessed at 12 wpi for the wild-type and at 9 wpi for the human derived infections, when the majority of mice were no longer excreting eggs. The numbers of worms recovered from the small intestine were few and there were no significant differences among the inbred or outbred groups of mice. We have found that C57BL/6J mice were the most susceptible to Brachylaima cribbi infection as assessed by excretion of eggs and provide a suitable model for a laboratory life cycle. PMID- 11880235 TI - Characterization of cDNAs encoding diacylglycerol acyltransferase from cultures of Brassica napus and sucrose-mediated induction of enzyme biosynthesis. AB - cDNAs encoding acyl-CoA:diacylglycerol acyltransferase (DGAT, EC 2.3.1.20), designated BnDGAT1 and BnDGAT2, were obtained from a microspore-derived cell suspension culture of oilseed rape (Brassica napus L. cv Jet Neuf). BnDGAT2 shares a very high level of identity with BnDGAT1, but is a smaller protein lacking the relatively hydrophilic N-terminal segment found in BnDGAT1. Both transcripts were produced in the cell suspension cultures and the cDNAs were functionally expressed in transformed yeast (Pichia pastoris) cells. Sucrose mediated changes in triacylglycerol (TAG) metabolism and expression of BnDGAT1 were examined in the cell suspension cultures following transfer of cells from media containing 6% (w/v) sucrose to media containing 14% sucrose. TAG content and DGAT activity of the cells increased transiently within the first 12 h after transfer (HAT). The rapid decline in TAG content observed at 12 HAT was inversely related to an increase in TAG lipase (EC 3.1.1.3) activity. The transient increases in TAG content and DGAT activity correlated with the elevated amounts of BnDGAT1 polypeptide. Transcript levels were also induced, but levels of mRNA encoding BnDGAT1 were not tightly correlated with DGAT activity and amount of polypeptide suggesting some control of expression at the post-transcriptional level. In general, the rapid changes in TAG content were closely associated with the changes in the activity of TAG-metabolizing enzymes and expression of BnDGAT1. PMID- 11880236 TI - A physicochemical investigation of two phosphatidylcholine/bile salt interfaces: implications for lipase activation. AB - Within the gastrointestinal tract ingested lipids are broken down into their constituent mono-acylglycerides and fatty acids by the enzyme family of lipases. In this study we have investigated the interfacial composition and structure of two phospholipid/bile salt (BS) systems that display significant differences in the duration of the lag phase of porcine pancreatic lipase kinetics. The interfacial tension of the single BSs, and their binary mixtures with phospholipid is reported at an n-tetradecane/water interface as a function of phospholipid mole fraction and total surfactant concentration. The structuring of the interface was probed by characterisation of the thin liquid film formation, thickness and stability. Lateral interactions were quantified by measurement of the diffusion coefficient of a probe fluorophore. We conclude that interfacial tension was not a factor in lag time duration as there was no significant difference in the minimum interfacial tension for the phosphatidylcholine (PC)/sodium taurocholate and the PC/sodium taurodeoxycholate system. No correlation was found between lag phase duration and the physiochemical properties of the interface, i.e. lateral diffusion, thin liquid film formation or interfacial tension. This is in agreement with our previous study that the lag time duration was directly related to the phospholipid content of the interface. PMID- 11880237 TI - Involvement of the GTPase Rho in the cellular uptake of low density lipoprotein by human skin fibroblasts. AB - Members of the Rho subfamily of small GTPases have been implicated in the regulation of endocytosis of ligand/receptor complexes localised to clathrin coated pits. In this paper, we investigated the role of Rho A in the post receptor regulation of cellular uptake and metabolism of native low density lipoprotein (LDL) by primary human skin fibroblasts. Incubations of cells with the selective Rho GTPase inhibitor C3-transferase (C3T) upregulated the binding, lysosomal degradation and cell accumulation of labelled LDL. The rate of internalisation of surface-bound LDL was also higher in C3T-treated cells. Single cell injections with C3T and dominant active V14Rho confirmed the negative regulation of LDL uptake by Rho. While cells injected with C3T internalised more 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (diI)-labelled LDL, diI-LDL internalisation was dramatically suppressed in cells injected with the constitutively active V14Rho. The negative regulation of LDL uptake by Rho appeared to be independent of changes in the actin cytoskeleton. An increasing number of naturally occurring toxins and serum factors have been shown to influence Rho GTPase signalling cascades. The herein described post-translational regulation of LDL internalisation may modulate cell events occurring subsequent to cellular lipoprotein uptake. PMID- 11880238 TI - Kidney microsomal 25- and 1alpha-hydroxylase in vitamin D metabolism: catalytic properties, molecular cloning, cellular localization and expression during development. AB - Both a 25-hydroxylation and a 1alpha-hydroxylation are necessary for the conversion of vitamin D(3) into the calcium-regulating hormone 1alpha,25 dihydroxyvitamin D(3). According to current knowledge, the hepatic mitochondrial cytochrome P450 (CYP) 27A and microsomal CYP2D25 are able to catalyze the former bioactivation step. Substantial 25-hydroxylase activity has also been demonstrated in kidney. This paper describes the molecular cloning and characterization of a microsomal vitamin D(3) 25- and 1alpha-hydroxylase in kidney. The enzyme purified from pig kidney and the recombinant enzyme expressed in COS cells catalyzed 25-hydroxylation of vitamin D(3) and 1alpha-hydroxyvitamin D(3) and, in addition, 1alpha-hydroxylation of 25-hydroxyvitamin D(3). The cDNA encodes a protein of 500 amino acids. Both the DNA sequence and the deduced peptide sequence of the renal enzyme are homologous with those of the hepatic vitamin D(3) 25-hydroxylase CYP2D25. Genomic Southern blot analysis suggested the presence of a single gene for CYP2D25 in the pig. Immunohistochemistry experiments indicated that CYP2D25 is expressed almost exclusively in the cells of cortical proximal tubules. The expression of CYP2D25 in kidney, but not in liver, was much higher in the adult pig than in the newborn. These findings indicate a tissue-specific developmental regulation of CYP2D25. The results from the current and previous studies on renal vitamin D hydroxylations imply that CYP2D25 has a biological role in kidney. PMID- 11880239 TI - Role of tyrosine kinase signaling in estrogen-induced LDL receptor gene expression in HepG2 cells. AB - The expression of the low-density lipoprotein receptor (LDL-r) gene is stimulated by estrogen in vivo, although its promoter does not contain a classical estrogen responsive element, suggesting an alternative mechanism of estrogen-regulated expression of this gene. The aim of this work was to assess whether estrogen stimulated transcription of the LDL-r gene depends on tyrosine kinase (TK) and protein kinase C (PKC) activation, both signaling pathways being activated by estrogen in vivo and in hepatoma cells. Therefore, in HepG2 cells cotransfected with estrogen receptor-alpha, estrogen-stimulated transcription of LDL-r-promoter reporter plasmid was analyzed in the absence and presence of TK and PKC inhibitors. The expression of LDL-r was also compared with the transcription of the complement gene, which contains a classical estrogen-responsive element sequence. Our results demonstrate that the induction of LDL-r expression by estrogen requires longer stimulation than that necessary for complement induction. Moreover, basal transcription of the LDL-r gene depends on PKC activity, while estrogen-stimulated activation of the LDL-r-promoter requires TK activity, pointing to a role of these non-classical estrogen-stimulated pathways in the transcriptional regulation of the LDL-r. PMID- 11880240 TI - The effect of conjugated linoleic acid on arachidonic acid metabolism and eicosanoid production in human saphenous vein endothelial cells. AB - The effects of a conjugated linoleic acid (CLA) mixture of single isomers (50:50, w/w, cis9,trans11:trans10,cis12) and the individual isomers on (a) the production of resting and calcium ionophore stimulated (14)C-eicosanoids and (b) the incorporation of (14)C-arachidonic acid (AA) into membrane phospholipids of human saphenous vein endothelial cells were investigated. The CLA mixture and the individual isomers were found to inhibit resting production of (14)C prostaglandin F(2a) by 50, 43 and 40%, respectively. A dose dependent inhibition of stimulated (14)C-prostaglandins was observed with the CLA mixture (IC(50) 100 microM). The cis9,trans11 and trans10,cis12 (50 microM) isomers individually inhibited the overall production of stimulated (14)C-prostaglandins (between 35 and 55% and 23 and 42%, respectively). When tested at a high concentration (100 microM), cis9,trans11 was found to inhibit eicosanoid production in contrast to trans10,cis12 that caused stimulation. The overall degree of (14)C-AA incorporation into membrane phospholipids of the CLA (mixture and individual isomers) treated cells was found to be lower than that of control cells and the cis9,trans11 isomer was found to increase the incorporation of (14)C-AA into phosphatidylcholine. Docosahexaenoic acid, eicosapentaenoic acid and linoleic acid did not alter the overall degree of incorporation of (14)C-AA. The results of this study suggest that both isomers inhibit eicosanoid production, and although trans10,cis12 exhibits pro-inflammatory activity at high concentrations, the CLA mixture maintains its beneficial anti-inflammatory action that contributes to its anti-carcinogenic and anti-atherogenic properties. PMID- 11880241 TI - Reduction in hepatic non-esterified fatty acid concentration after long-term treatment with atorvastatin lowers hepatic triglyceride synthesis and its secretion in sucrose-fed rats. AB - The mechanism by which atorvastatin lowers plasma triglyceride (TG) levels is mainly through a decrease in hepatic TG secretion. However, it is not clear why atorvastatin, which does not inhibit TG synthesis in vitro, decreases hepatic TG secretion without a prospective increase in hepatic TG concentration. For the investigation of the mechanisms that underlie the hypotriglyceridemic effects of atorvastatin, we characterized the effect of either a single or an 11 day administration of atorvastatin in sucrose-induced hypertriglyceridemic rats. Atorvastatin (30 mg/kg p.o.) strongly decreased the rate of both very-low-density lipoprotein (VLDL)-TG and VLDL-apolipoprotein B secretion. The inhibitor also decreased hepatic TG concentration. Hepatic TG synthesis activity was also decreased by atorvastatin, and its activity was correlated with both hepatic and plasma TG concentration. There was also a strong correlation between the hepatic TG synthesis and hepatic non-esterified fatty acid (NEFA) concentration (r(2)=0.815). These effects required chronic administration of the inhibitor and were not observed by acute treatment. Repeated administration of atorvastatin also strongly reduced hepatic acyl-coenzyme A synthase mRNA levels. These results suggest that the reduced hepatic NEFA most likely lowers hepatic TG synthesis and TG secretion in sucrose-fed hypertriglyceridemic rats. PMID- 11880242 TI - Kinetic analyses of liver phosphatidylcholine and phosphatidylethanolamine biosynthesis using (13)C NMR spectroscopy. AB - Choline and ethanolamine are substrates for de novo synthesis of phosphatidylcholine (PtdC) and phosphatidylethanolamine (PtdE) through the CDP choline and CDP-ethanolamine pathways. In liver, PtdE can also be converted to PtdC by PtdE N-methyltransferase (PEMT). We investigated these kinetics in rat liver during a 60 min infusion with (13)C-labeled choline and ethanolamine. NMR analyses of liver extracts provided concentrations and (13)C enrichments of phosphocholine (Pcho), phosphoethanolamine (Peth), PtdC, and PtdE. Kinetic models showed that the de novo and PEMT pathways are 'channeled' processes. The intermediary metabolites directly derived from exogenous choline and ethanolamine do not completely mix with the intracellular pools, but are preferentially used for phospholipid synthesis. Of the newly synthesized PtdC, about 70% was derived de novo and 30% was by PEMT. PtdC and PtdE de novo syntheses displayed different kinetics. A simple model assuming constant fluxes yielded a modest fit to the data; allowing upregulated fluxes significantly improved the fit. The ethanolamine-to-Peth flux exceeded choline-to-Pcho, and the rate of PtdE synthesis (1.04 micromol/h/g liver) was 2-3 times greater than that of PtdC de novo synthesis. The metabolic pathway information provided by these studies makes the NMR method superior to earlier radioisotope studies. PMID- 11880243 TI - Micelle formation of sodium deoxycholate and sodium ursodeoxycholate (part 1). AB - Micellization of sodium deoxycholate (NaDC) and sodium ursodeoxycholate (NaUDC) was studied for the critical micelle concentration (CMC), the micelle aggregation number, and the degree of counterion binding to micelle, where sodium cholate (NaC) was used as a reference. The fluorescence probe technique of pyrene was employed to determine accurately the CMC values for the bile salts, which indicated that a certain concentration range of CMC and a stepwise aggregation for micellization were reasonable. The temperature dependences of micellization for NaDC and NaUDC were studied at 288.2, 298.2, 308.2, and 318.2 K by aqueous solubility change with solution pH. Aggregations of the bile salt anions were analyzed using the stepwise association model and found to grow in size with increasing concentration, which confirmed that the mass action model worked quite well. The average aggregation number was found to be 2.5 (NaUDC) and 10.5 (NaDC) at the concentration of 20 mM and at 308.2 K. The aggregation number determined by static light scattering also agreed well with those by the solubility method in the order of size: NaUDC 25 +/- 3 mM, n = 4). Despite a 1,000-fold decrease in Ca(2+) affinity in D542N, Ca(2+) permeation properties and the Ca(2+)-to-Ba(2+) conductance ratio remained comparable to values for wild-type ECaC1. Together, our observations suggest that D542 plays a critical role in Ca(2+) affinity but not in Ca(2+) permeation in ECaC1. PMID- 11880256 TI - Expression, localization, and functional evaluation of CFTR in bovine corneal endothelial cells. AB - HCO-dependent fluid secretion by the corneal endothelium controls corneal hydration and maintains corneal transparency. Recently, it has been shown that mRNA for the cystic fibrosis transmembrane conductance regulator (CFTR) is expressed in the corneal endothelium; however, protein expression, functional localization, and a possible role in HCO transport have not been reported. Immunoblotting for CFTR showed a single band at approximately 170 kDa for both freshly isolated and primary cultures of bovine corneal endothelial cells. Indirect immunofluorescence confocal microscopy indicated that CFTR locates to the apical membrane. Relative changes in apical and basolateral chloride permeability were estimated by measuring the rate of fluorescence quenching of the halide-sensitive indicator 6-methoxy-N-ethylquinolinium iodide during Cl(-) influx in the absence and presence of forskolin (FSK). Apical and basolateral Cl( ) permeability increased 10- and 3-fold, respectively, in the presence of 50 microM FSK. FSK-activated apical chloride permeability was unaffected by H(2)DIDs (250 microM); however, 5-nitro-2-(3-phenylpropyl-amino)benzoic acid (NPPB; 50 microM) and glibenclamide (100 microM ) inhibited activated Cl(-) fluxes by 45% and 30%, respectively. FSK-activated basolateral Cl(-) permeability was insensitive to NPPB, glibenclamide, or furosemide but was inhibited 80% by H(2)DIDS. HCO permeability was estimated by measuring changes in intracellular pH in response to quickly lowering bath [HCO]. FSK (50 microM) increased apical HCO permeability by twofold, which was inhibited 42% by NPPB and 65% by glibenclamide. Basolateral HCO permeability was unaffected by FSK. Genistein (50 microM) significantly increased apical HCO and Cl(minus sign) permeability by 1.8 and 16-fold, respectively. When 50 microM genistein was combined with 50 microM FSK, there was no further increase in Cl(-) permeability; however, HCO permeability was reduced to the control level. In summary, we conclude that CFTR is present in the apical membrane of bovine corneal endothelium and could contribute to transendothelial Cl(-) and HCO transport. Furthermore, there is a cAMP-activated Cl(-) pathway on the basolateral membrane that is not CFTR. PMID- 11880257 TI - Redox states of type 1 ryanodine receptor alter Ca(2+) release channel response to modulators. AB - The type 1 ryanodine receptor (RyR1) from rabbit skeletal muscle displayed two distinct degrees of response to cytoplasmic Ca(2+) [high- and low-open probability (P(o)) channels]. Here, we examined the effects of adenine nucleotides and caffeine on these channels and their modulations by sulfhydryl reagents. High-P(o) channels showed biphasic Ca(2+) dependence and were activated by adenine nucleotides and caffeine. Unexpectedly, low-P(o) channels did not respond to either modulator. The addition of a reducing reagent, dithiothreitol, to the cis side converted the high-P(o) channel to a state similar to that of the low-P(o) channel. Treatment with p-chloromercuriphenylsulfonic acid (pCMPS) transformed low-P(o) channels to a high-P(o) channel-like state with stimulation by beta,gamma-methylene-ATP and caffeine. In experiments under redox control using glutathione buffers, shift of the cis potential toward the oxidative state activated the low-P(o) channel, similar to that of the high-P(o) or the pCMPS treated channel, whereas reductive changes inactivated the high-P(o) channel. Changes in trans redox potential, in contrast, did not affect channel activity of either channel. In all experiments, channels with higher P(o) were stimulated to a great extent by modulators, but ones with lower P(o) were unresponsive. These results suggest that redox states of critical sulfhydryls located on the cytoplasmic side of the RyR1 may alter both gating properties of the channel and responsiveness to channel modulators. PMID- 11880258 TI - cAMP-dependent protein kinase and proliferation differ in normal and polycystic kidney epithelia. AB - Developmental control of cell proliferation is crucial, and abnormal principal cell proliferation may contribute to cystogenesis in polycystic kidney disease. This study investigates roles of cAMP and its primary effector, cAMP-dependent protein kinase (protein kinase A; PKA), in control of cell proliferation in filter-grown noncystic (NC) and cystic (CY)-derived principal cell cultures. These cultures had similar cAMP pathway characteristics upstream of PKA subunit distribution but differed in predicted PKA subtype distribution. Functionally, cultures were proliferative before polarization, with constitutively higher proliferation in CY cultures. NC cultures achieved levels similar to those of CY cultures on pharmacological manipulation of cAMP production or PKA activation or inhibition of PKA subtype I activity. Inhibition of overall PKA activity, or of PKA subtype II anchoring, diminished cAMP/PKA-mediated proliferation in NC cultures but had no effect on CY cultures. Polarized CY monolayers remained proliferative, but NC monolayers lost responsiveness. No large proliferation changes resulted from treatments of polarized cultures; however, polarized NC and CY cultures differed in poststimulation handling of PKA catalytic and type IIalpha regulatory subunits. Our results support PKA subtype regulation of prepolarization proliferation in NC principal cells and altered regulation of PKA in CY cells and suggest that differences at or downstream of PKA can contribute to altered proliferation in a developmental renal disease. PMID- 11880259 TI - Dual effect of fluid shear stress on volume-regulated anion current in bovine aortic endothelial cells. AB - The key mechanism responsible for maintaining cell volume homeostasis is activation of volume-regulated anion current (VRAC). The role of hemodynamic shear stress in the regulation of VRAC in bovine aortic endothelial cells was investigated. We showed that acute changes in shear stress have a biphasic effect on the development of VRAC. A shear stress step from a background flow (0.1 dyn/cm(2)) to 1 dyn/cm(2) enhanced VRAC activation induced by an osmotic challenge. Flow alone, in the absence of osmotic stress, did not induce VRAC activation. Increasing the shear stress to 3 dyn/cm(2), however, resulted in only a transient increase of VRAC activity followed by an inhibitory phase during which VRAC was gradually suppressed. When shear stress was increased further (5 10 dyn/cm(2)), the current was immediately strongly suppressed. Suppression of VRAC was observed both in cells challenged osmotically and in cells that developed spontaneous VRAC under isotonic conditions. Our findings suggest that shear stress is an important factor in regulating the ability of vascular endothelial cells to maintain volume homeostasis. PMID- 11880260 TI - Secretory modulation of basolateral membrane inwardly rectified K(+) channel in guinea pig distal colonic crypts. AB - Cell-attached recordings revealed K(+) channel activity in basolateral membranes of guinea pig distal colonic crypts. Inwardly rectified currents were apparent with a pipette solution containing 140 mM K(+). Single-channel conductance (gamma) was 9 pS at the resting membrane potential. Another inward rectifier with gamma of 19 pS was observed occasionally. At a holding potential of -80 mV, gamma was 21 and 41 pS, respectively. Identity as K(+) channels was confirmed after patch excision by changing the bath ion composition. From reversal potentials, relative permeability of Na(+) over K(+) (P(Na)/P(K)) was 0.02 +/- 0.02, with P(Rb)/P(K) = 1.1 and P(Cl)/P(K) < 0.03. Spontaneous open probability (P(o)) of the 9-pS inward rectifier ((gp)K(ir)) was voltage independent in cell-attached patches. Both a low (P(o) = 0.09 +/- 0.01) and a moderate (P(o) = 0.41 +/- 0.01) activity mode were observed. Excision moved (gp)K(ir) to the medium activity mode; P(o) of (gp)K(ir) was independent of bath Ca(2+) activity and bath acidification. Addition of Cl(-) and K(+) secretagogues altered P(o) of (gp)K(ir). Forskolin or carbachol (10 microM) activated the small-conductance (gp)K(ir) in quiescent patches and increased P(o) in low-activity patches. K(+) secretagogues, either epinephrine (5 microM) or prostaglandin E(2) (100 nM), decreased P(o) of (gp)K(ir) in active patches. This (gp)K(ir) may be involved in electrogenic secretion of Cl(minus sign) and K(+) across the colonic epithelium, which requires a large basolateral membrane K(+) conductance during maximal Cl(-) secretion and, presumably, a lower K(+) conductance during primary electrogenic K(+) secretion. PMID- 11880261 TI - Acid secretion and proton conductance in human airway epithelium. AB - Acid secretion and proton conductive pathways across primary human airway surface epithelial cultures were investigated with the pH stat method in Ussing chambers and by single cell patch clamping. Cultures showed a basal proton secretion of 0.17 +/- 0.04 micromol.h(-1).cm(-2), and mucosal pH equilibrated at 6.85 +/- 0.26. Addition of histamine or ATP to the mucosal medium increased proton secretion by 0.27 +/- 0.09 and 0.24 +/- 0.09 micromol.h(-1).cm(-2), respectively. Addition of mast cells to the mucosal medium of airway cultures similarly activated proton secretion. Stimulated proton secretion was similar in cultures bathed mucosally with either NaCl Ringer or ion-free mannitol solutions. Proton secretion was potently blocked by mucosal ZnCl(2) and was unaffected by mucosal bafilomycin A(1), Sch-28080, or ouabain. Mucosal amiloride blocked proton secretion in tissues that showed large amiloride-sensitive potentials. Proton secretion was sensitive to the application of transepithelial current and showed outward rectification. In whole cell patch-clamp recordings a strongly outward rectifying, zinc-sensitive, depolarization-activated proton conductance was identified with an average chord conductance of 9.2 +/- 3.8 pS/pF (at 0 mV and a pH 5.3-to-pH 7.3 gradient). We suggest that inflammatory processes activate proton secretion by the airway epithelium and acidify the airway surface liquid. PMID- 11880262 TI - Characteristics of hyperpolarization-activated cation currents in portal vein smooth muscle cells. AB - Voltage-clamp studies of freshly isolated smooth muscle cells from rabbit portal vein revealed the existence of a time-dependent cation current evoked by membrane hyperpolarization (termed I(h)). Both the rate of activation and the amplitude of I(h) were enhanced by membrane hyperpolarization. Half-maximal activation of I(h) was about -105 mV with conventional whole cell and -80 mV when the perforated patch technique was used. In current clamp, injection of hyperpolarizing current produced a marked depolarizing "sag" followed by rebound depolarization. Activation of I(h) was augmented by an increase in the extracellular K(+) concentration and was blocked rapidly by externally applied Cs(+) (1-5 mM). The bradycardic agent ZD-7288 (10 microM), a selective inhibitor of I(h), produced a characteristically slow inhibition of the portal vein I(h). The depolarizing sag recorded in current clamp was also abolished by application of 5 mM Cs(+). Cs(+) significantly decreased the frequency of spontaneous contractions in both whole rat portal vein and rabbit portal vein segments. Multiplex RT-PCR of rabbit portal vein myocytes using primers derived from existing genes for hyperpolarization-activated cation channels (HCN1-4) revealed the existence of cDNA clones corresponding to HCN2, 3, and 4. The present study shows that portal vein myocytes contain genes shown to encode for hyperpolarization-activated channels and exhibit an endogenous current with characteristics similar to I(h) in other cell types. This conductance appears to determine, in part, the rhythmicity of this vessel. PMID- 11880263 TI - CaM kinase II-dependent activation of tyrosine kinases and ERK1/2 in vascular smooth muscle. AB - In vascular smooth muscle (VSM) and many other cells, G protein receptor-coupled activation of mitogen-activated protein kinases has been linked, in part, to increases in free intracellular Ca(2+). Previously, we demonstrated that ionomycin-, angiotensin II-, and thrombin-induced activation of extracellular signal-regulated kinase (ERK)1/2 in VSM cells was attenuated by pretreatment with KN-93, a selective inhibitor of the multifunctional Ca(2+)/calmodulin-dependent protein kinase (CaM kinase II). In the present study, we show that the Ca(2+) dependent pathway leading to activation of ERK1/2 is preceded by nonreceptor proline-rich tyrosine kinase (PYK2) activation and epidermal growth factor (EGF) receptor tyrosine phosphorylation and is attenuated by inhibitors of src family kinases or the EGF receptor tyrosine kinase. Furthermore, we demonstrate that pretreatment with KN-93 or a CaM kinase II inhibitor peptide inhibits Ca(2+) dependent PYK2 activation and EGF receptor tyrosine phosphorylation in response to ionomycin, ATP, and platelet-derived growth factor but has no effect on phorbol 12,13-dibutyrate- or EGF-induced responses. The results implicate CaM kinase II as an intermediate in the Ca(2+)/calmodulin-dependent activation of PYK2. PMID- 11880264 TI - Expression of nitric oxide synthase isoforms and detection of nitric oxide in rat placenta. AB - Nitric oxide (NO) production in the rat placenta was monitored and quantified by electron paramagnetic resonance (EPR) spectroscopy with hemoglobin and an Fe-N (dithiocarboxy)sarcosine (DTCS) complex as NO-trapping reagents. Expression of nitric oxide synthase (NOS) isoforms was also examined by quantitative RT-PCR analysis. The EPR spectrum of the placenta with hemoglobin trapping showed a three-line hyperfine structure (g = 2.008 and a = 1.66-mT). The EPR signal was diminished after the placenta was homogenized or the NOS inhibitor L-NAME was administered to pregnant rats. Therefore, the specific signal was definitely identified as being derived from endogenous NO spin-trapped by hemoglobin, and the EPR spectrum showed that the NO adduct existed as a pentacoordinate alpha-NO heme species. The EPR spectrum of the placenta with Fe-DTCS trapping showed a triplet signal (g = 2.038) derived from an NO-Fe-DTCS complex. The height of the triplet signal did not vary significantly with gestational stage during the last few days of gestation. At the gestational stages examined, the level of NOS II mRNA expression was significantly higher than that of NOS III mRNA. NOS II expression in term (day 21.5) placenta was significantly increased compared with that in preterm (day 19.5) placenta (P < 0.01, n = 4 or 5). These results suggest that NOS II is the predominant producer of NO in the placenta and that NOS II generated NO plays significant roles in the maintenance of placental functions immediately before birth. PMID- 11880265 TI - Enhancement of L-type Ca(2+) current from neonatal mouse ventricular myocytes by constitutively active PKC-betaII. AB - The cardiac L-type calcium current (I(Ca)) can be modified by activation of protein kinase C (PKC). However, the effect of PKC activation on I(Ca) is still controversial. Some studies have shown a decrease in current, whereas other studies have reported a biphasic effect (an increase followed by a decrease in current or vice versa). A possible explanation for the conflicting results is that several isoforms of PKC with opposing effects on I(Ca) were activated simultaneously. Here, we examined the influence of a single PKC isoform (PKC betaII) on L-type calcium channels in isolation from other cardiac isoforms, using a transgenic mouse that conditionally expresses PKC-betaII. Ventricular cardiac myocytes were isolated from newborn mice and examined for expression of the transgene using single cell RT-PCR after I(Ca) recording. Cells expressing PKC-betaII showed a twofold increase in nifedipine-sensitive I(Ca). The PKC betaII antagonist LY-379196 returned I(Ca) amplitude to levels found in non-PKC betaII-expressing myocytes. The increase in I(Ca) was independent of Ca(v)1.2 subunit mRNA levels as determined by quantitative RT-PCR. Thus these data demonstrate that PKC-beta is a potent modulator of cardiac L-type calcium channels and that this specific isoform increases I(Ca) in neonatal ventricular myocytes. PMID- 11880266 TI - Effect of HUVEC on human osteoprogenitor cell differentiation needs heterotypic gap junction communication. AB - Bone development and remodeling depend on complex interactions between bone forming osteoblasts and other cells present within the bone microenvironment, particularly vascular endothelial cells that may be pivotal members of a complex interactive communication network in bone. Our aim was to investigate the interaction between human umbilical vein endothelial cells (HUVEC) and human bone marrow stromal cells (HBMSC). Cell differentiation analysis performed with different cell culture models revealed that alkaline phosphatase activity and type I collagen synthesis were increased only by the direct contact of HUVEC with HBMSC. This "juxtacrine signaling" could involve a number of different heterotypic connexions that require adhesion molecules or gap junctions. A dye coupling assay with Lucifer yellow demonstrated a functional coupling between HUVEC and HBMSC. Immunocytochemistry revealed that connexin43 (Cx43), a specific gap junction protein, is expressed not only in HBMSC but also in the endothelial cell network and that these two cell types can communicate via a gap junctional channel constituted at least by Cx43. Moreover, functional inhibition of the gap junction by 18alpha-glycyrrhetinic acid treatment or inhibition of Cx43 synthesis with oligodeoxyribonucleotide antisense decreased the effect of HUVEC cocultures on HBMSC differentiation. This stimulation could be mediated by the intercellular diffusion of signaling molecules that permeate the junctional channel. PMID- 11880268 TI - Permeant but not impermeant divalent cations enhance activation of nondesensitizing alpha(7) nicotinic receptors. AB - Neuronal alpha(7) nicotinic acetylcholine receptors (nAChRs) are permeable to Ca(2+) and other divalent cations. We characterized the modulation of the pharmacological properties of nondesensitizing mutant (L(247)T and S(240)T/L(247)T) alpha(7) nAChRs by permeant (Ca(2+), Ba(2+), and Sr(2+)) and impermeant (Cd(2+) and Zn(2+)) divalent cations. alpha(7) receptors were expressed in Xenopus oocytes and studied with two-electrode voltage clamp. Extracellular permeant divalent cations increased the potency and maximal efficacy of ACh, whereas impermeant divalent cations decreased potency and maximal efficacy. The antagonist dihydro-beta-erythroidine (DHbetaE) was a strong partial agonist of L(247)T and S(240)T/L(247)T alpha(7) receptors in the presence of divalent cations but was a weak partial agonist in the presence of impermeant divalent cations. Mutation of the "intermediate ring" glutamates (E(237)A) in L(247)T alpha(7) nAChRs eliminated Ca(2+) conductance but did not alter the Ca(2+)-dependent increase in ACh potency, suggesting that site(s) required for modulation are on the extracellular side of the intermediate ring. The difference between permeant and impermeant divalent cations suggests that sites within the pore are important for modulation by divalent cations. PMID- 11880269 TI - Distribution of ClC-2 chloride channel in rat and human epithelial tissues. AB - The ubiquitous ClC-2 Cl(-) channel is thought to contribute to epithelial Cl(-) secretion, but the distribution of the ClC-2 protein in human epithelia has not been investigated. We have studied the distribution of ClC-2 in adult human and rat intestine and airways by immunoblotting and confocal microscopy. In the rat, ClC-2 was present in the lateral membranes of villus enterocytes and was predominant at the basolateral membranes of luminal colon enterocytes. The expression pattern of ClC-2 in the human intestine differed significantly, because ClC-2 was mainly detected in a supranuclear compartment of colon cells. We found significant expression of ClC-2 at the apex of ciliated cells in both rat and human airways. These results show that the distribution of ClC-2 in airways is consistent with participation of ClC-2 channels in Cl(-) secretion and indicate that extrapolation of results from studies of ClC-2 function in rat intestine to human intestine is not straightforward. PMID- 11880267 TI - Cellular distribution of parchorin, a chloride intracellular channel-related protein, in various tissues. AB - The cellular distribution of parchorin, a new chloride intracellular channel family member, was investigated in rabbit tissues by immunohistochemistry using an antibody recognizing the sequence containing a parchorin-specific repeat. Parchorin preferentially resides in the epithelium of the ducts of the lacrymal, parotid, submandibular, and mammary glands and the pancreas, prostate, and testis. In the trachea and lung, parchorin was found in the airway epithelium and type II alveolar cells. In the kidney, parchorin was distributed mainly from the thick ascending limb to the distal convoluted tubule. In the eye, both pigment and nonpigment epithelia of the ciliary body were positive, whereas only the pigment epithelium was positive in the retina. Parchorin was also present in the cochlea and semicircular canal. The amount of parchorin in the gastric mucosa, but not in the submandibular glands, increased after weaning. In the mammary gland, parchorin expression was greater in a lactating rabbit (1 wk after delivery) compared with a pregnant (3 wk) rabbit. The cellular distribution and changes in expression indicate that parchorin plays an important role, possibly in chloride transport, in the cells that create an ion gradient for water movement. PMID- 11880270 TI - Phosphorylation of the salivary Na(+)-K(+)-2Cl(-) cotransporter. AB - We studied the phosphorylation of the secretory Na(+)-K(+)-2Cl(-) cotransporter (NKCC1) in rat parotid acinar cells. We have previously shown that NKCC1 activity in these cells is dramatically upregulated in response to beta-adrenergic stimulation and that this upregulation correlates with NKCC1 phosphorylation, possibly due to protein kinase A (PKA). We show here that when ATP is added to purified acinar basolateral membranes (BLM), NKCC1 is phosphorylated as a result of membrane-associated protein kinase activity. Additional NKCC1 phosphorylation is seen when PKA is added to BLMs, but our data indicate that this is due to an effect of PKA on endogenous membrane kinase or phosphatase activities, rather than its direct phosphorylation of NKCC1. Also, phosphopeptide mapping demonstrates that these phosphorylations do not take place at the site associated with the upregulation of NKCC1 by beta-adrenergic stimulation. However, this upregulatory phosphorylation can be mimicked by the addition of cAMP to permeabilized acini, and this effect can be blocked by a specific PKA inhibitor. These latter results provide good evidence that PKA is indeed involved in the upregulatory phosphorylation of NKCC1 and suggest that an additional factor present in the acinar cell but absent from isolated membranes is required to bring about the phosphorylation. PMID- 11880271 TI - Regulation of COX-2 expression in human intestinal myofibroblasts: mechanisms of IL-1-mediated induction. AB - Elevated mucosal interleukin-1 (IL-1) levels are frequently seen during acute and chronic intestinal inflammation, and IL-1 neutralization lessens the severity of inflammation. One major effect of IL-1 is the increased release of eicosanoid mediators via induction of cyclooxygenase-2 (COX-2). One site of COX-2-derived prostaglandin synthesis during acute and chronic intestinal inflammation is the intestinal myofibroblast. COX-2 expression has also been documented in these cells in colonic neoplasms. Thus an understanding of the regulation of COX-2 expression in human intestinal myofibroblasts is important. As an initial step toward this goal we have characterized IL-1alpha signaling pathways that induce COX-2 expression in cultured human intestinal myofibroblasts. IL-1 treatment resulted in a dramatic transcriptional induction of COX-2 gene expression. Activation of nuclear factor-kappaB (NF-kappaB), extracellular signal-regulated protein kinase (ERK), p38, and protein kinase C (PKC) signaling pathways was each necessary for optimal COX-2 induction. In contrast to what occurs in other cell types, including other myofibroblasts such as renal mesangial cells, PKC inhibition did not prevent IL-1-induced NF-kappaB or mitogen activated protein kinase/ stress-activated protein kinase activation, suggesting a novel role for PKC isoforms during this process. The stimulatory effects of PKC, NF-kappaB, ERK 1/2, and presumably c-Jun NH(2)-terminal kinase activation were exerted at the transcriptional level, whereas p38 activation resulted in increased stability of the COX-2 message. We conclude that, in intestinal myofibroblasts, IL-1-mediated induction of COX-2 expression is a complex process that requires input from multiple signaling pathways. Each parallel pathway acts in relative autonomy, the sum of their actions culminating in a dramatic increase in COX-2 transcription and message stability. PMID- 11880272 TI - Influence of myosin isoforms on contractile properties of intact muscle fibers from Rana pipiens. AB - The myosin heavy chain (MHC) and myosin light chain (MLC) isoforms in skeletal muscle of Rana pipiens have been well characterized. We measured the force velocity (F-V) properties of single intact fast-twitch fibers from R. pipiens that contained MHC types 1 or 2 (MHC1 or MHC2) or coexpressed MHC1 and MHC2 isoforms. Velocities were measured between two surface markers that spanned most of the fiber length. MHC and MLC isoform content was quantified after mechanics analysis by SDS-PAGE. Maximal shortening velocity (V(max)) and velocity at half maximal tension (V(P 50)) increased with percentage of MHC1 (%MHC1). Maximal specific tension (P(o)/CSA, where P(o) is isometric tension and CSA is fiber cross-sectional area) and maximal mechanical power (W(max)) also increased with %MHC1. MHC concentration was not significantly correlated with %MHC1, indicating that the influence of %MHC1 on P(o)/CSA and W(max) was due to intrinsic differences between MHC isoforms and not to concentration. The MLC3-to-MLC1 ratio was not significantly correlated with V(max), V(P 50), P(o)/CSA, or W(max). These data demonstrate the powerful relationship between MHC isoforms and F-V properties of the two most common R. pipiens fiber types. PMID- 11880273 TI - Sustained norepinephrine contraction in the rat portal vein is lost when Ca(2+) is replaced with Sr(2+). AB - Agonist-induced activation of smooth muscle involves a rise in intracellular Ca(2+) concentration and sensitization of myosin light chain phosphorylation to Ca(2+). Sr(2+) can enter through Ca(2+) channels, be sequestered and released from sarcoplasmic reticulum, and replace Ca(2+) in activation of myosin light chain phosphorylation. Sr(2+) cannot replace Ca(2+) in facilitation of agonist activated Ca(2+)-dependent nonselective cation channels. It is not known whether Sr(2+) can replace Ca(2+) in small G protein-mediated sensitization of phosphorylation. To explore mechanisms involved in alpha-receptor-activated contractions in smooth muscle, effects of replacing Ca(2+) with Sr(2+) were examined in rat portal vein. Norepinephrine (NE) at >3.0 x 10(-7) M in the presence of Ca(2+) resulted in a strong sustained contraction, whereas this sustained component was absent in the presence of Sr(2+); only the amplitude of phasic contractions increased. Pretreatment with low (approximately 0.05 mM) free Ca(2+) followed by 2.5 mM Sr(2+) resulted in a sustained component of the NE response. In beta-escin-permeabilized preparations, phenylephrine in the presence of GTP or guanosine 5'-O-(3-thiotriphosphate) alone induced sensitization to Sr(2+). In conclusion, a Ca(2+)-regulated membrane/channel process is required for the sustained component of NE responses in rat portal vein. Sensitization alone is not responsible for the sustained phase of the NE contraction. PMID- 11880274 TI - Proadipogenic effect of leptin on rat preadipocytes in vitro: activation of MAPK and STAT3 signaling pathways. AB - Because leptin has recently been shown to induce proliferation and/or differentiation of different cell types through different pathways, the aim of the present study was to investigate, in vitro, the influence of leptin on adipogenesis in rat preadipocytes. A prerequisite to this study was to identify leptin receptors (Ob-Ra and Ob-Rb) in preadipocytes from femoral subcutaneous fat. We observed that expressions of Ob-Ra and Ob-Rb increase during adipogenesis. Furthermore, leptin induces an increase of p42/p44 mitogen activated protein kinase phosphorylated isoforms in both confluent and differentiated preadipocytes and of STAT3 phosphorylation only in confluent preadipocytes. Moreover, exposure to leptin promoted activator protein-1 complex DNA binding activity in confluent preadipocytes. Finally, exposure of primary cultured preadipocytes from the subcutaneous area to leptin (10 nM) resulted in an increased proliferation ([(3)H]thymidine incorporation and cell counting) and differentiation (glycerol-3-phosphate dehydrogenase activity and mRNA levels of lipoprotein lipase, peroxisome proliferator-activated receptor-gamma2, and c fos). Altogether, these results indicate that, in vitro at least, leptin through its functional receptors exerts a proadipogenic action in subcutaneous preadipocytes. PMID- 11880275 TI - An extracellular sulfhydryl group modulates background Na(+) conductance and cytosolic Ca(2+) in pituitary cells. AB - Treatment of GH(3) pituitary cells with p-chloromercurybenzenesulfonate (PCMBS) increased the cytosolic Ca(2+) concentration ([Ca(2+)](i)). This effect was reversed by dithiothreitol and blocked by L-type Ca(2+) channel antagonists or Na(+) removal. PCMBS increased membrane conductance and depolarized the plasma membrane. Apart from minor effects on K(+) and Ca(2+) channels, PCMBS increased (6 times at -80 mV) an inward Na(+) current whose properties were similar to those of a background Na(+) conductance (BNC) described previously, necessary for generation of spontaneous electrical activity. In rat lactotropes and somatotropes in primary culture, PCMBS also produced a Na(+)-dependent [Ca(2+)](i) increase, whereas little or no effect was observed in thyrotropes, corticotropes, and gonadotropes. The Na(+) conductance elicited by PCMBS in somatotropes seemed to be the same as that stimulated by the hypothalamic growth hormone (GH)-releasing hormone, which regulates membrane excitability and GH secretion. The BNC studied here could play a physiological role, regulating excitability and spontaneous activity, and explains satisfactorily the [Ca(2+)](i)-increasing actions of the mercurials reported previously in several excitable tissues. PMID- 11880276 TI - Laminins and TGF-beta maintain cell polarity and functionality of human gastric glandular epithelium. AB - The human gastric glandular epithelium produces a gastric lipase enzyme (HGL) that plays an important role in digestion of dietary triglycerides. To assess the involvement of extracellular matrix components and transforming growth factor beta1 (TGF-beta1) in the regulation of this enzymic function, normal gastric epithelial cells were cultured on collagen type I, Matrigel, and laminins (LN)-1 and -2 with or without TGF-beta1. Epithelial morphology and HGL expression were evaluated using microscopy techniques, enzymic assays, Western blot, Northern hybridization, and RT-PCR. A correlation was observed between the cell polarity status and the level of HGL expression. TGF-beta1 alone or individual matrix components stimulated cell spreading and caused a downfall of HGL activity and mRNA. By contrast, Matrigel preserved the morphological features of differentiated epithelial cells and maintained HGL expression. The combination of LNs with TGF-beta1 (two constituents of Matrigel) exerted similar beneficial effects on epithelial cell polarity and evoked a 10-fold increase of HGL levels that was blunted by a neutralizing antibody against the alpha(2)-integrin subunit and by mitogen-activated protein kinase (MAPK) inhibitors PD-98059 (p42/p44) or SB-203580 (p38). This investigation demonstrates for the first time that a powerful synergism between a growth factor and basement membrane LNs positively influences cell polarity and functionality of the human gastric glandular epithelium through an activation of the alpha(2)beta(1)-integrin and effectors of two MAPK pathways. PMID- 11880277 TI - Activation of K(+) channels and increased migration of differentiated intestinal epithelial cells after wounding. AB - Early mucosal restitution occurs by epithelial cell migration to reseal superficial wounds after injury. Differentiated intestinal epithelial cells induced by forced expression of the Cdx2 gene migrate over the wounded edge much faster than undifferentiated parental cells in an in vitro model. This study determined whether these differentiated intestinal epithelial cells exhibit increased migration by altering voltage-gated K(+) (Kv) channel expression and cytosolic free Ca(2+) concentration ([Ca(2+)](cyt)). Stable Cdx2-transfected IEC 6 cells (IEC-Cdx2L1) with highly differentiated phenotype expressed higher basal levels of Kv1.1 and Kv1.5 mRNAs and proteins than parental IEC-6 cells. Neither IEC-Cdx2L1 cells nor parental IEC-6 cells expressed voltage-dependent Ca(2+) channels. The increased expression of Kv channels in differentiated IEC-Cdx2L1 cells was associated with an increase in whole cell K(+) currents, membrane hyperpolarization, and a rise in [Ca(2+)](cyt). The migration rates in differentiated IEC-Cdx2L1 cells were about four times those of parental IEC-6 cells. Inhibition of Kv channel expression by polyamine depletion decreased [Ca(2+)](cyt), reduced myosin stress fibers, and inhibited cell migration. Elevation of [Ca(2+)](cyt) by ionomycin promoted myosin II stress fiber formation and increased cell migration. These results suggest that increased migration of differentiated intestinal epithelial cells is mediated, at least partially, by increasing Kv channel activity and Ca(2+) influx during restitution. PMID- 11880278 TI - Satellite cell proliferation and differentiation during postnatal growth of porcine skeletal muscle. AB - Age-related changes in satellite cell proliferation and differentiation during rapid growth of porcine skeletal muscle were examined. Satellite cells were isolated from hindlimb muscles of pigs at 1, 7, 14, and 21 wk of age (4 animals/age group). Satellite cells were separated from cellular debris by using Percoll gradient centrifugation and were adsorbed to glass coverslips for fluorescent immunostaining. Positive staining for neural cell adhesion molecule (NCAM) distinguished satellite cells from nonmyogenic cells. The proportion of NCAM-positive cells (satellite cells) in isolates decreased from 1 to 7 wk of age. Greater than 77% of NCAM-positive cells were proliferating cell nuclear antigen positive at all ages studied. Myogenin-positive satellite cells decreased from 30% at 1 wk to 14% at 7 wk of age and remained at constant levels thereafter. These data indicate that a high percentage of satellite cells remain proliferative during rapid postnatal muscle growth. The reduced proportion of myogenin-positive cells during growth may reflect a decrease in the proportion of differentiating satellite cells or accelerated incorporation of myogenin-positive cells into myofibers. PMID- 11880279 TI - Nongastric H-K-ATPase in rodent prostate: lobe-specific expression and apical localization. AB - The molecular basis of active ion transport in secretory glands such as the prostate is not well characterized. Rat nongastric H-K-ATPase is expressed at high levels in distal colon surface cell apical membranes and thus is referred to as "colonic." Here we show that the ATPase is expressed in rodent prostate complex in a lobe-specific manner. RT-PCR and Western blot analyses indicate that rat nongastric H-K-ATPase alpha-subunit (alpha(ng)) mRNA and protein are present in coagulating gland (anterior prostate) and lateral and dorsal prostate and absent from ventral lobe, whereas Na-K-ATPase alpha-subunit is present in all lobes. RT-PCR analysis shows that Na-K-ATPase alpha(4) and alpha(3) and gastric H K-ATPase alpha-subunit are not present in significant amounts in all prostate lobes. Relatively low levels of Na-K-ATPase alpha(2) were found in lateral, dorsal, and anterior lobes. alpha(ng) protein expression is anteriodorsolateral: highest in coagulating gland, somewhat lower in dorsal lobe, and even lower in lateral lobe. Na-K-ATPase protein abundance has the reverse order: expression in ventral lobe is higher than in coagulating gland. alpha(ng) protein abundance is higher in coagulating gland than distal colon membranes. Immunohistochemistry shows that in rat and mouse coagulating gland epithelium alpha(ng) protein has an apical polarization and Na-K-ATPase alpha(1) is localized in basolateral membranes. The presence of nongastric H-K-ATPase in rodent prostate apical membranes may indicate its involvement in potassium concentration regulation in secretions of these glands. PMID- 11880280 TI - Differential roles of ICAM-1 and E-selectin in polymorphonuclear leukocyte induced angiogenesis. AB - Ets-1, which stimulates metalloproteinase gene transcription, has a key role in angiogenesis. We first examined whether activated polymorphonuclear leukocytes (PMNs) enhanced angiogenesis through the induction of Ets-1. Addition of activated PMNs to endothelial cells stimulated both in vitro angiogenesis in collagen gel and Ets-1 expression. Both angiogenesis and Ets-1 expression induced by PMNs were reduced by ets-1 antisense oligonucleotide, suggesting that Ets-1 is an important factor in PMN-induced angiogenesis. Although intercellular adhesion molecule (ICAM)-1 and E-selectin are involved in PMN-induced angiogenesis, the mechanisms underlying their roles in angiogenesis have yet to be elucidated. PMN induced Ets-1 expression was reduced by a monoclonal antibody against ICAM-1 but not E-selectin despite the inhibition of PMN-induced angiogenesis by both antibodies. Moreover, the stimulation of angiogenesis by H(2)O(2) without PMNs was inhibited by a monoclonal antibody to E-selectin but not ICAM-1. These findings suggested that ICAM-1 in endothelial cells may act as a signaling receptor to induce Ets-1 expression, whereas E-selectin seems to function in the formation of tubelike structures in vascular endothelial cell cultures. PMID- 11880282 TI - Heterologous desensitization of EGF receptors and PDGF receptors by sequestration in caveolae. AB - Epidermal growth factor (EGF) and platelet-derived growth factor (PDGF) receptors have been reported to signal via caveolin-containing membranes called caveolae. In contrast, others report that EGF and PDGF receptors are exclusively associated with caveolin-devoid membranes called rafts. Our subcellular fractionation and coimmunoprecipitation studies demonstrate that, in the absence of ligand, EGF and PDGF receptors are associated with rafts. However, in the presence of ligand, EGF and PDGF receptors transiently associate with caveolae. Surprisingly, pretreatment of cells with EGF prevents PDGF-dependent phosphorylation of PDGF receptors and extracellular signal-regulated kinase (ERK) 1/2 kinase activation. Furthermore, cells pretreated with PDGF prevent EGF-dependent phosphorylation of EGF receptors and ERK1/2 kinase activation. Radioligand binding studies demonstrate that incubation of cells with EGF or PDGF causes both EGF and PDGF receptors to be reversibly sequestered from the extracellular space. Experiments with methyl-beta-cyclodextrin, filipin, and antisense caveolin-1 demonstrate that sequestration of the receptors is dependent on cholesterol and caveolin-1. We conclude that ligand-induced stimulation of EGF or PDGF receptors can cause the heterologous desensitization of the other receptor by sequestration in cholesterol-rich, caveolin-containing membranes or caveolae. PMID- 11880281 TI - Role of reactive oxygen species and NAD(P)H oxidase in alpha(1)-adrenoceptor signaling in adult rat cardiac myocytes. AB - We recently reported that alpha(1)-adrenoceptor (alpha(1)-AR) stimulation induces hypertrophy via activation of the mitogen/extracellular signal-regulated kinase (MEK) 1/2-extracellular signal-regulated kinase (ERK) 1/2 pathway and generates reactive oxygen species (ROS) in adult rat ventricular myocytes (ARVM). Here we investigate the intracellular source of ROS in ARVM and the mechanism by which ROS activate hypertrophic signaling after alpha(1)-AR stimulation. Pretreatment of ARVM with the ROS scavenger Mn(III)terakis(1-methyl-4-pyridyl) porphyrin pentachloride (MnTMPyP) completely inhibited the alpha(1)-AR-stimulated activation of Ras-MEK1/2-ERK1/2. Direct addition of H(2)O(2) or the superoxide generator menadione activated ERK1/2, which is also prevented by MnTMPyP pretreatment. We found that ARVM express gp91(phox), p22(phox), p67(phox), and p47(phox), four major components of NAD(P)H oxidase, and that alpha(1)-AR stimulated ERK1/2 activation was blocked by four structurally unrelated inhibitors of NAD(P)H oxidase [diphenyleneiodonium, phenylarsine oxide, 4-(2 aminoethyl)benzenesulfonyl fluoride, and cadmium]. Conversely, inhibitors for other potential ROS-producing systems, including mitochondrial electron transport chain, nitric oxide synthase, xanthine oxidase, and cyclooxygenase, had no effect on alpha(1)-AR-stimulated ERK1/2 activation. Taken together, our results show that ventricular myocytes express components of an NAD(P)H oxidase that appear to be involved in alpha(1)-AR-stimulated hypertrophic signaling via ROS-mediated activation of Ras-MEK1/2-ERK1/2. PMID- 11880284 TI - Na transport proteins are expressed by rat alveolar epithelial type I cells. AB - Despite a presumptive role for type I (AT1) cells in alveolar epithelial transport, specific Na transporters have not previously been localized to these cells. To evaluate expression of Na transporters in AT1 cells, double labeling immunofluorescence microscopy was utilized in whole lung and in cytocentrifuged preparations of partially purified alveolar epithelial cells (AEC). Expression of Na pump subunit isoforms and the alpha-subunit of the rat (r) epithelial Na channel (alpha-ENaC) was evaluated in isolated AT1 cells identified by their immunoreactivity with AT1 cell-specific antibody markers (VIIIB2 and/or anti aquaporin-5) and lack of reactivity with antibodies specific for AT2 cells (anti surfactant protein A) or leukocytes (anti-leukocyte common antigen). Expression of the Na pump alpha(1)-subunit in AEC was assessed in situ. Na pump subunit isoform and alpha-rENaC expression was also evaluated by RT-PCR in highly purified (approximately 95%) AT1 cell preparations. Labeling of isolated AT1 cells with anti-alpha(1) and anti-beta(1) Na pump subunit and anti-alpha-rENaC antibodies was detected, while reactivity with anti-alpha(2) Na pump subunit antibody was absent. AT1 cells in situ were reactive with anti-alpha(1) Na pump subunit antibody. Na pump alpha(1)- and beta(1)- (but not alpha(2)-) subunits and alpha-rENaC were detected in highly purified AT1 cells by RT-PCR. These data demonstrate that AT1 cells express Na pump and Na channel proteins, supporting a role for AT1 cells in active transalveolar epithelial Na transport. PMID- 11880285 TI - Beta-adrenergic regulation of amiloride-sensitive lung sodium channels. AB - We investigated the mechanism by which cAMP increases sodium transport in lung epithelial cells. Alveolar type II (ATII) cells have two types of amiloride sensitive, cation channels: a nonselective cation channel (NSC) and a highly selective channel (HSC). Exposure of ATII cells to cAMP, beta-adrenergic agonists, or other agents that increase adenylyl cyclase activity increased activity of both channel types, albeit by different mechanisms. NSC open probability (P(o)) increased severalfold when exposed to terbutaline, isoproterenol, forskolin, or cAMP analogs without any change in NSC number. In contrast, terbutaline increased HSC number with no significant change in HSC P(o). For both channels, the effect of terbutaline was blocked by propranolol and H-89, suggesting a protein kinase A (PKA) requirement for beta-adrenergic-induced changes in channel activity. Terbutaline increased cAMP levels in ATII cells, but intracellular calcium also increased. Calcium sequestration with BAPTA blocked beta-adrenergic-induced increases in NSC P(o) but did not alter HSC activity. These observations suggest that beta-adrenergic stimulation increases intracellular cAMP and activates PKA. PKA increases HSC number and increases intracellular calcium. The increase in calcium increases NSC P(o). Thus increased cAMP levels are likely to increase lung sodium transport regardless of which channel type is present. PMID- 11880286 TI - Beta-adrenoceptor-mediated control of apical membrane conductive properties in fetal distal lung epithelia. AB - Distal lung epithelial cells isolated from fetal rats were cultured (48 h) on permeable supports so that transepithelial ion transport could be quantified electrometrically. Unstimulated cells generated a short-circuit current (I(sc)) that was inhibited (~80%) by apical amiloride. The current is thus due, predominantly, to the absorption of Na(+) from the apical solution. Isoprenaline increased the amiloride-sensitive I(sc) about twofold. Experiments in which apical membrane Na(+) currents were monitored in basolaterally permeabilized cells showed that this was accompanied by a rise in apical Na(+) conductance (G(Na(+))). Isoprenaline also increased apical Cl- conductance (G(Cl-)) by activating an anion channel species sensitive to glibenclamide but unaffected by 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS). The isoprenaline evoked changes in G(Na(+)) and G(Cl(minus sign)) could account for the changes in I(sc) observed in intact cells. Glibenclamide had no effect upon the isoprenaline evoked stimulation of I(sc) or G(Na(+)) demonstrating that the rise in G(Cl-) is not essential to the stimulation of Na(+) transport. PMID- 11880287 TI - Glucocorticoid-stimulated lung epithelial Na(+) transport is associated with regulated ENaC and sgk1 expression. AB - H441 cells, a bronchiolar epithelial cell line, develop a glucocorticoid regulated amiloride-sensitive Na(+) transport pathway on permeable supports (R. Sayegh, S. D. Auerbach, X. Li, R. Loftus, R. Husted, J. B. Stokes, and C. P. Thomas. J Biol Chem 274: 12431-12437, 1999). To understand its molecular basis, we examined the effect of glucocorticoids (GC) on epithelial Na(+) channel (ENaC) alpha, -beta, and -gamma and sgk1 expression and determined the biophysical properties of Na(+) channels in these cells. GC stimulated the expression of ENac alpha, -beta, and -gamma and sgk1 mRNA, with the first effect seen by 1 h. These effects were abolished by actinomycin D, but not by cycloheximide, indicating a direct stimulatory effect on ENaC and sgk1 mRNA synthesis. The GC effect on transcription of ENaC-alpha mRNA was accompanied by a significant increase in ENaC-alpha protein levels. GC also stimulated ENaC-alpha, -beta, and -gamma and sgk1 mRNA expression in A549 cells, an alveolar type II cell line. To determine the biophysical properties of the Na(+) channel, single-channel currents were recorded from cell-attached H441 membranes. An Na(+)-selective channel with slow kinetics and a slope conductance of 10.8 pS was noted, properties similar to ENaC alpha, -beta, and -gamma expressed in Xenopus laevis oocytes. These experiments indicate that amiloride-sensitive Na(+) transport is mediated through classic ENaC channels in human lung epithelia and that GC-regulated Na(+) transport is accompanied by increased transcription of each of the component subunits and sgk1. PMID- 11880288 TI - Ca(2+)-dependent stimulation of alveolar fluid clearance in near-term fetal guinea pigs. AB - We investigated the importance of changes in intracellular Ca(2+) concentration ([Ca(2+)](i)) for amiloride-sensitive alveolar fluid clearance (AFC) in late gestational guinea pigs. Fetal guinea pigs of 61, 68, and 69 days (term) gestation were investigated under normal conditions and after oxytocin-induced preterm labor. AFC or alveolar fluid secretion was measured using an impermeable tracer technique. At 61 days gestation there was net secretion of fluid into the lungs, and at birth the lungs cleared 49 +/- 7% of the instilled fluid volume over 1 h. Induction of preterm labor with oxytocin induced AFC at 61 days gestation. When present, AFC was inhibited or reversed to net fluid secretion by amiloride (10(-3) M). Inhibition of membrane Ca(2+) channels by verapamil (10(-4) M) or depletion of intracellular Ca(2+) by thapsigargin (10(-5) M) reduced AFC when net AFC was evident. Amiloride lacked an inhibitory effect on AFC when instilled with verapamil or thapsigargin. The results indicate that AFC via amiloride-sensitive pathways develops during late gestation, and that inducing preterm labor precociously may activate such pathways. Our results suggest that Ca(2+) may act as a second messenger in mediating catecholamine-stimulated AFC. PMID- 11880289 TI - cAMP regulation of Cl(-) and HCO(-)(3) secretion across rat fetal distal lung epithelial cells. AB - We isolated and cultured fetal distal lung epithelial (FDLE) cells from 17- to 19 day rat fetuses and assayed for anion secretion in Ussing chambers. With symmetrical Ringer solutions, basal short-circuit currents (I(sc)) and transepithelial resistances were 7.9 +/- 0.5 microA/cm(2) and 1,018 +/- 73 Omega.cm(2), respectively (means +/- SE; n = 12). Apical amiloride (10 microM) inhibited basal I(sc) by approximately 50%. Subsequent addition of forskolin (10 microM) increased I(sc) from 3.9 +/- 0.63 microA/cm(2) to 7.51 +/- 0.2 microA/cm(2) (n = 12). Basolateral bumetanide (100 microM) decreased forskolin stimulated I(sc) from 7.51 +/- 0.2 microA/cm(2) to 5.62 +/- 0.53, whereas basolateral 4,4'-dinitrostilbene-2,2'-disulfonate (5 mM), an inhibitor of HCO secretion, blocked the remaining I(sc). Forskolin addition evoked currents of similar fractional magnitudes in symmetrical Cl(-)- or HCO(-)(3)-free solutions; however, no response was seen using HCO(-)(3)- and Cl(-)-free solutions. The forskolin-stimulated I(sc) was inhibited by glibenclamide but not apical DIDS. Glibenclamide also blocked forskolin-induced I(sc) across monolayers having nystatin-permeablized basolateral membranes. Immunolocalization studies were consistent with the expression of cystic fibrosis transmembrane conductance regulator (CFTR) protein in FDLE cells. In aggregate, these findings indicate the presence of cAMP-activated Cl(-) and HCO(-)(3) secretion across rat FDLE cells mediated via CFTR. PMID- 11880290 TI - Hypoxia decreases active Na transport across primary rat alveolar epithelial cell monolayers. AB - Hypoxia has been reported to inhibit activity and expression of ion transporters of alveolar epithelial cells. This study extended those observations by investigating the mechanisms underlying inhibition of active Na transport across primary cultured adult rat alveolar epithelial cell monolayers grown on polycarbonate filters. Cell monolayers were exposed to normoxia and hypoxia (1.5% and 5% O(2), 5% CO(2)), and resultant changes in bioelectric properties [i.e., short-circuit current (I(sc)) and transepithelial resistance (R(t))] were measured in Ussing chambers. Results showed that I(sc) decreased with duration of exposure to hypoxia, while relatively little change was observed for R(t). In normoxia, amiloride inhibited approximately 70% of I(sc). The amiloride-sensitive portion of I(sc) decreased over time of exposure to hypoxia, whereas the magnitude of the amiloride-insensitive portion of I(sc) was not affected. Na pump capacity measured after permeabilization of the apical plasma membrane with amphotericin B decreased in monolayers exposed to 1.5% O(2) for 24 h, as did the capacity of amiloride-sensitive Na uptake measured after imposing an apical to basolateral Na gradient and permeabilization of the basolateral membrane. These results demonstrate that exposure to hypoxia inhibits alveolar epithelial Na reabsorption by reducing the rates of both apical amiloride-sensitive Na entry and basolateral Na extrusion. PMID- 11880291 TI - Prolonged isoproterenol infusion impairs the ability of beta(2)-agonists to increase alveolar liquid clearance. AB - We determined if prolonged isoproterenol (Iso) infusion in rats impaired the ability of the beta(2)-adrenergic agonist terbutaline to increase alveolar liquid clearance (ALC). We infused rats with Iso (at rates of 4, 40, or 400 microg.kg( 1).h(-1)) or vehicle (0.001 N HCl) for 48 h using subcutaneously implanted miniosmotic pumps. After this time, the rats were anesthetized, and ALC was determined (by mass-balance after instillation of Ringer lactate containing albumin into the lungs) under baseline conditions and after terbutaline administration. Baseline and terbutaline-stimulated ALC in vehicle-infused rats averaged, respectively, 19.6 +/- 1.2% (SE) and 44.7 +/- 1.5%/h. The ability of terbutaline to increase ALC was eliminated at 400 microg.kg(-1).h(-1)Iso, inhibited by 26% at 40 microg.kg(-1).h(-1) Iso, and was not affected by 4 microg.kg(-1).h(-1) Iso. beta-adrenergic receptor (betaAR) density of freshly isolated alveolar epithelial type II (ATII) cells from Iso-infused rats was reduced by the 40 and 400 microg.kg(-1).h(-1) infusion rates. These data demonstrate that prolonged exposure to beta-agonists can impair the ability of beta(2)-agonists to stimulate ALC and produce ATII cell betaAR downregulation. PMID- 11880292 TI - Polarity of alveolar epithelial cell acid-base permeability. AB - We investigated acid-base permeability properties of electrically resistive monolayers of alveolar epithelial cells (AEC) grown in primary culture. AEC monolayers were grown on tissue culture-treated polycarbonate filters. Filters were mounted in a partitioned cuvette containing two fluid compartments (apical and basolateral) separated by the adherent monolayer, cells were loaded with the pH-sensitive dye 2',7'-bis(2-carboxyethyl)-5(6)-carboxyfluorescein, and intracellular pH was determined. Monolayers in HCO-free Na(+) buffer (140 mM Na(+), 6 mM HEPES, pH 7.4) maintained a transepithelial pH gradient between the two fluid compartments over 30 min. Replacement of apical fluid by acidic (6.4) or basic (8.0) buffer resulted in minimal changes in intracellular pH. Replacement of basolateral fluid by acidic or basic buffer resulted in transmembrane proton fluxes and intracellular acidification or alkalinization. Intracellular alkalinization was blocked > or =80% by 100 microM dimethylamiloride, an inhibitor of Na(+)/H(+) exchange, whereas acidification was not affected by a series of acid/base transport inhibitors. Additional experiments in which AEC monolayers were grown in the presence of acidic (6.4) or basic (8.0) medium revealed differential effects on bioelectric properties depending on whether extracellular pH was altered in apical or basolateral fluid compartments bathing the cells. Acid exposure reduced (and base exposure increased) short-circuit current from the basolateral side; apical exposure did not affect short-circuit current in either case. We conclude that AEC monolayers are relatively impermeable to transepithelial acid/base fluxes, primarily because of impermeability of intercellular junctions and of the apical, rather than basolateral, cell membrane. The principal basolateral acid exit pathway observed under these experimental conditions is Na(+)/H(+) exchange, whereas proton uptake into cells occurs across the basolateral cell membrane by a different, undetermined mechanism. These results are consistent with the ability of the alveolar epithelium to maintain an apical-to-basolateral (air space-to-blood) pH gradient in situ. PMID- 11880293 TI - Pattern of NOS2 and NOS3 mRNA expression in human A549 cells and primary cultured AEC II. AB - The human alveolar type II epithelium-like cell line A549 expresses nitric oxide synthase type 2 (NOS2), but not NOS3, and produces nitric oxide (NO) upon appropriate stimulation. However, relatively little is known regarding the NOS2 and NOS3 expression of type II human alveolar epithelial cells (AEC II) in primary culture. We detected NOS3 mRNA in freshly isolated AEC II and after 24 h of culture. NOS3 mRNA levels were much higher in AEC II cultured for 24 h with or without interferon-gamma, interleukin-1beta, and tumor necrosis factor-alpha, compared with freshly isolated cells. Cytokine stimulation did not change the NOS3 mRNA expression level in AEC II compared with unstimulated cells. NOS3 protein expression was verified by Western blot, and measuring nitrate/nitrite revealed that the protein is active. In contrast, neither NOS2 mRNA nor protein could be detected in freshly isolated, unstimulated or cytokine-stimulated human AEC II in 24- or 72-h primary cultures, whereas A549 cells expressed NOS2 message and protein upon stimulation with proinflammatory cytokines. In situ hybridization confirmed that AEC II express NOS3, but not NOS2 mRNA in vivo. These data demonstrate that there are significant differences between primary AEC II and A549 cells in NOS mRNA expression pattern. PMID- 11880294 TI - Carbon monoxide promotes hypoxic pulmonary vascular remodeling. AB - CO is a biologically active gas that produces cellular effects by multiple mechanisms. Because cellular binding of CO by heme proteins is increased in hypoxia, we tested the hypothesis that CO interferes with hypoxic pulmonary vascular remodeling in vivo. Rats were exposed to inspired CO (50 parts/million) at sea level or 18,000 ft of altitude [hypobaric hypoxia (HH)], and changes in vessel morphometry and pulmonary pressure-flow relationships were compared with controls. Vascular cell single strand DNA (ssDNA) and proliferating cell nuclear antigen (PCNA) were assessed, and changes in gene and protein expression of smooth muscle alpha-actin (sm-alpha-actin), beta-actin, and heme oxygenase-1 (HO 1) were evaluated by Western analysis, RT-PCR, and immunohistochemistry. After 21 days of HH, vascular pressure at constant flow and vessel wall thickness increased and lumen diameter of small arteries decreased significantly. The presence of CO, however, further increased both pulmonary vascular resistance (PVR) and the number of small muscular vessels compared with HH alone. CO + HH also increased vascular PCNA and nuclear ssDNA expression compared with hypoxia, suggesting accelerated cell turnover. CO in hypoxia downregulated sm-alpha-actin and strongly upregulated beta-actin. CO also increased lung HO activity and HO-1 mRNA and protein expression in small pulmonary arteries during hypoxia. These data indicate an overall propensity of CO in HH to promote vascular remodeling and increase PVR in vivo. PMID- 11880295 TI - Exaggerated hypoxic pulmonary hypertension in endothelin B receptor-deficient rats. AB - Mechanisms by which endothelin (ET)-1 mediates chronic pulmonary hypertension remain incompletely understood. Although activation of the ET type A (ET(A)) receptor causes vasoconstriction, stimulation of ET type B (ET(B)) receptors can elicit vasodilation or vasoconstriction. We hypothesized that the ET(B) receptor attenuates the development of hypoxic pulmonary hypertension and studied a genetic rat model of ET(B) receptor deficiency (transgenic sl/sl). After 3 wk of severe hypoxia, the transgenic sl/sl pulmonary vasculature lacked expression of mRNA for the ET(B) receptor and developed exaggerated pulmonary hypertension that was characterized by elevated pulmonary arterial pressure, diminished cardiac output, and increased total pulmonary resistance. Plasma ET-1 was fivefold higher in transgenic sl/sl rats than in transgenic controls. Although mRNA for prepro-ET 1 was not different, mRNA for ET-converting enzyme-1 was higher in transgenic sl/sl than in transgenic control lungs. Hypertensive lungs of sl/sl rats also produced less nitric oxide metabolites and 6-ketoprostaglandin F(1alpha), a metabolite of prostacyclin, than transgenic controls. These findings suggest that the ET(B) receptor plays a protective role in the pulmonary hypertensive response to chronic hypoxia. PMID- 11880296 TI - Angiotensin receptor subtype AT(1) mediates alveolar epithelial cell apoptosis in response to ANG II. AB - Previous work from this laboratory demonstrated induction of apoptosis in lung alveolar epithelial cells (AEC) by purified angiotensin II (ANG II) and expression of mRNAs for both ANG II receptor subtypes AT(1) and AT(2) (Wang R, Zagariya A, Ibarra-Sunga O, Gidea C, Ang E, Deshmukh S, Chaudhary G, Baraboutis J, Filippatos G, and Uhal BD. Am J Physiol Lung Cell Mol Physiol 276: L885-L889, 1999.). The present study was designed to determine the ANG II receptor subtype mediating AEC apoptosis in response to ANG II. Apoptosis was induced with purified ANG II applied to the human lung AEC-derived carcinoma cell line A549 or to primary AEC isolated from Wistar rats. In both cell types, the AT(1)-selective receptor antagonists L-158809 or losartan inhibited ANG II-induced apoptosis by 90% at concentrations of 10(-8) M and 10(-7) M, respectively. The inhibition was concentration dependent with IC(50) of 10(-12) M and 10(-11) M on the primary rat AEC. In contrast, the AT(2)-selective antagonists PD-123319 or PD-126055 could not block ANG II-induced apoptosis in either cell type. In primary rat AEC, apoptosis in response to ANG II was blunted in a dose-dependent manner by the protein kinase C inhibitor chelerythrine but not by the tyrosine phosphatase inhibitor sodium orthovanadate. Together, these data indicate that AEC apoptosis in response to ANG II is mediated by receptor subtype AT(1), despite the expression of mRNAs for both AT(1) and AT(2). PMID- 11880297 TI - Role of extracellular superoxide dismutase in bleomycin-induced pulmonary fibrosis. AB - Bleomycin administration results in well-described intracellular oxidative stress that can lead to pulmonary fibrosis. The role of alveolar interstitial antioxidants in this model is unknown. Extracellular superoxide dismutase (EC SOD) is the primary endogenous extracellular antioxidant enzyme and is abundant in the lung. We hypothesized that EC-SOD plays an important role in attenuating bleomycin-induced lung injury. Two weeks after intratracheal bleomycin administration, we found that wild-type mice induced a 106 +/- 25% increase in lung EC-SOD. Immunohistochemical staining revealed that a large increase in EC SOD occurred in injured lung. Using mice that overexpress EC-SOD specifically in the lung, we found a 53 +/- 14% reduction in bleomycin-induced lung injury assessed histologically and a 17 +/- 6% reduction in lung collagen content 2 wk after bleomycin administration. We conclude that EC-SOD plays an important role in reducing the magnitude of lung injury from extracellular free radicals after bleomycin administration. PMID- 11880298 TI - Anoxia-induced apoptosis occurs through a mitochondria-dependent pathway in lung epithelial cells. AB - The intracellular signaling pathways that control O(2) deprivation (anoxia) induced apoptosis have not been fully defined in lung epithelial cells. We show here that the lung epithelial cell line A549 releases cytochrome c and activates caspase-9 followed by DNA fragmentation and plasma membrane breakage in response to anoxia. The antiapoptotic protein Bcl-X(L) prevented the anoxia-induced cell death by inhibiting the release of cytochrome c and caspase-9 activation. A549 cells devoid of mitochondrial DNA (rho(o)-cells) and lacking a functional electron transport chain were resistant to anoxia-induced apoptosis. A549 cells preconditioned with either hypoxia (1.5% O(2)) or tumor necrosis factor-alpha, which activated the transcription factors hypoxia-inducible factor-1 or nuclear factor-kappaB, respectively, did not provide protection from anoxia-induced cell death. These results indicate that A549 cells require a functional electron transport chain and the release of cytochrome c for anoxia-induced apoptosis. PMID- 11880299 TI - Pulmonary-specific expression of tumor necrosis factor-alpha alters surfactant lipid metabolism. AB - Tumor necrosis factor (TNF)-alpha is a major cytokine implicated in inducing acute and chronic lung injury, conditions associated with surfactant phosphatidylcholine (PtdCho) deficiency. Acutely, TNF-alpha decreases PtdCho synthesis but stimulates surfactant secretion. To investigate chronic effects of TNF-alpha, we investigated PtdCho metabolism in a murine transgenic model exhibiting lung-specific TNF-alpha overexpression. Compared with controls, TNF alpha transgenic mice exhibited a discordant pattern of PtdCho metabolism, with a decrease in PtdCho and disaturated PtdCho (DSPtdCho) content in the lung, but increased levels in alveolar lavage. Transgenics had lower activities and increased immunoreactive levels of cytidylyltransferase (CCT), a key PtdCho biosynthetic enzyme. Ceramide, a CCT inhibitor, was elevated, and linoleic acid, a CCT activator, was decreased in transgenics. Radiolabeling studies revealed that alveolar reuptake of DSPtdCho was significantly decreased in transgenic mice. These observations suggest that chronic expression of TNF-alpha results in a complex pattern of PtdCho metabolism where elevated lavage PtdCho may originate from alveolar inflammatory cells, decreased surfactant reuptake, or altered surfactant secretion. Reduced parenchymal PtdCho synthesis appears to be attributed to CCT enzyme that is physiologically inactivated by ceramide or by diminished availability of activating lipids. PMID- 11880300 TI - Effect of surfactant on pulmonary expression of type IIA PLA(2) in an animal model of acute lung injury. AB - We previously showed that the seminatural surfactant Curosurf inhibits the in vitro synthesis of secretory type IIA phospholipase A(2) (sPLA(2)-IIA) in alveolar macrophages (AM). These cells are the main source of sPLA(2)-IIA in a guinea pig model of lipopolysaccharide (LPS)-induced acute lung injury (ALI). Here, we investigate the effect of Curosurf on the pulmonary synthesis of sPLA(2) IIA in this ALI model. Our results showed that intratracheal administration of LPS (330 microg/kg) induced an increase in pulmonary expression of sPLA(2)-IIA, which was inhibited when animals received Curosurf (16 mg/guinea pig) 30 min or 8 h after LPS instillation. When AM were isolated from LPS-treated animals and cultured in conditioned medium, they expressed higher levels of sPLA(2)-IIA than AM from saline-treated animals. This ex vivo sPLA(2)-IIA expression was significantly reduced when guinea pigs received Curosurf 30 min after LPS instillation. Finally, we examined the effect of Curosurf on pulmonary inflammation measured 8 or 24 h after LPS administration. Curosurf instillation 30 min or 8 h after LPS reversed the increase in tumor necrosis factor-alpha expression, polymorphonuclear cell extravasation, and protein concentration in bronchoalveolar lavage fluids. Curosurf also decreased the bronchial reactivity induced by LPS. We conclude that Curosurf inhibits the pulmonary expression of sPLA(2)-IIA and exhibits palliative anti-inflammatory effects in an animal model of ALI. PMID- 11880301 TI - Identification of Pseudomonas aeruginosa flagellin as an adhesin for Muc1 mucin. AB - We reported previously that Muc1 mucin on the epithelial cell surface is an adhesion site for Pseudomonas aeruginosa (Lillehoj EP, Hyun SW, Kim BT, Zhang XG, Lee DI, Rowland S, and Kim KC. Am J Physiol Lung Cell Mol Physiol 280: L181-L187, 2001). The present study was designed to identify the adhesin(s) responsible for bacterial binding to Muc1 mucin using genetic and biochemical approaches. Chinese hamster ovary (CHO) cells stably transfected with a Muc1 cDNA (CHO-Muc1) or empty plasmid (CHO-X) were compared for adhesion of P. aeruginosa strain PAK. Our results showed that 1) wild-type PAK and isogenic mutant strains lacking pili (PAK/NP) or flagella cap protein (PAK/fliD) demonstrated significantly increased binding to CHO-Muc1 cells, whereas flagellin-deficient (PAK/fliC) bacteria were no more adherent to CHO-Muc1 than CHO-X cells, and 2) P. aeruginosa adhesion was blocked by pretreatment of bacteria with antibody to flagellin or pretreatment of CHO-Muc1 cells with purified flagellin. We conclude that flagellin is an adhesin of P. aeruginosa responsible for its binding to Muc1 mucin on the epithelial cell surface. PMID- 11880302 TI - Cell-specific involvement of HNF-1beta in alpha(1)-antitrypsin gene expression in human respiratory epithelial cells. AB - The synergistic action of hepatocyte nuclear factor (HNF)-1alpha and HNF-4 plays an important role in expression of the alpha(1)-antitrypsin (alpha(1)-AT) gene in human hepatic and intestinal epithelial cells. Recent studies have indicated that the alpha(1)-AT gene is also expressed in human pulmonary alveolar epithelial cells, a potentially important local site of the lung antiprotease defense. In this study, we examined the possibility that alpha(1)-AT gene expression in a human pulmonary epithelial cell line H441 was also directed by the synergistic action of HNF-1alpha and HNF-4 and/or by the action of HNF-3, which has been shown to play a dominant role in gene expression in H441 cells. The results show that alpha(1)-AT gene expression in H441 cells is predominantly driven by HNF 1beta, even though HNF-1beta has no effect on alpha(1)-AT gene expression in human hepatic Hep G2 and human intestinal epithelial Caco-2 cell lines. Expression of alpha(1)-AT and HNF-1beta was also demonstrated in primary cultures of human respiratory epithelial cells. HNF-4 has no effect on alpha(1)-AT gene expression in H441 cells, even when it is cotransfected with HNF-1beta or HNF 1alpha. HNF-3 by itself has little effect on alpha(1)-AT gene expression in H441, Hep G2, or Caco-2 cells but tends to have an upregulating effect when cotransfected with HNF-1 in Hep G2 and Caco-2 cells. These results indicate the unique involvement of HNF-1beta in alpha(1)-AT gene expression in a cell line and primary cultures derived from human respiratory epithelium. PMID- 11880303 TI - Morpho-functional analysis of lung tissue in mild interstitial edema. AB - Mild pulmonary interstitial edema was shown to cause fragmentation of interstitial matrix proteoglycans. We therefore studied compartmental fluid accumulation by light and electron microscopy on lungs of anesthetized rabbits fixed in situ by vascular perfusion after 0.5 ml.kg(-1).min(-1) iv saline infusion for 180 min causing approximately 6% increase in lung weight. Morphometry showed that a relevant portion (44%) of extravascular fluid is detected early in the alveolar septa, 85% of this fluid accumulating in the thick portion of the air-blood barrier. The arithmetic mean thickness of the barrier increased in interstitial edema from 1.06 +/- 0.05 (SE) to 1.33 +/- 0.06 microm. The harmonic mean thickness increased from 0.6 +/- 0.03 to 0.86 +/- 0.07 microm, mostly due to thickening of the thin portion causing an increase in gas diffusion resistance. Despite some structural damage, the air-blood barrier displays a relatively high structural resistance providing a safety factor against the development of severe edema. It is suggested that the increase in extra-alveolar perivascular space occurs as a consequence of fluid accumulation in the air-blood barrier. PMID- 11880304 TI - Allergic inflammation-induced neuropeptide production in rapidly adapting afferent nerves in guinea pig airways. AB - In the vagal-sensory system, neuropeptides such as substance P and calcitonin gene-related peptide (CGRP) are synthesized nearly exclusively in small-diameter nociceptive type C-fiber neurons. By definition, these neurons are designed to respond to noxious or tissue-damaging stimuli. A common feature of visceral inflammation is the elevation in production of sensory neuropeptides. Little is known, however, about the physiological characteristics of vagal sensory neurons induced by inflammation to produce substance P. In the present study, we show that allergic inflammation of guinea pig airways leads to the induction of substance P and CGRP production in large-diameter vagal sensory neurons. Electrophysiological and anatomical evidence reveals that the peripheral terminals of these neurons are low-threshold Adelta mechanosensors that are insensitive to nociceptive stimuli such as capsaicin and bradykinin. Thus inflammation causes a qualitative change in chemical coding of vagal primary afferent neurons. The results support the hypothesis that during an inflammatory reaction, sensory neuropeptide release from primary afferent nerve endings in the periphery and central nervous system does not require noxious or nociceptive stimuli but may also occur simply as a result of stimulation of low-threshold mechanosensors. This may contribute to the heightened reflex physiology and pain that often accompany inflammatory diseases. PMID- 11880305 TI - NADPH oxidase promotes NF-kappaB activation and proliferation in human airway smooth muscle. AB - Evidence is rapidly accumulating that low-activity-reduced nicotinamide adenine dinucleotide phosphate (NADPH) oxidases homologous to that in phagocytic cells generate reactive oxygen species as signaling intermediates in both endothelium and vascular smooth muscle. We therefore explored the possibility of such an oxidase regulating growth of airway smooth muscle (AWSM). Proliferation of human AWSM cells in culture was inhibited by the antioxidants catalase and N acetylcysteine, and by the flavoprotein inhibitor diphenylene iodonium (DPI). Membranes prepared from human AWSM cells generated superoxide anion (O) measured by superoxide dismutase-inhibitable lucigenin chemiluminescence, with a distinct preference for NADPH instead of reduced nicotinamide adenine dinucleotide as substrate. Chemiluminescence was also inhibited by DPI, suggesting the presence of a flavoprotein containing oxidase generating O as a signaling molecule for cell growth. Examination of human AWSM cells by reverse transcriptase-polymerase chain reaction consistently demonstrated transcripts with sequences identical to those reported for p22(phox). Transfection with p22(phox) antisense oligonucleotides reduced human AWSM proliferation. Inhibition of NADPH oxidase activity with DPI prevented serum-induced activation of nuclear factor-kappaB (NF kappaB), and overexpression of a superrepressor form of the NF-kappaB inhibitor IkappaBalpha significantly reduced human AWSM growth. These findings suggest that an NADPH oxidase containing p22(phox) regulates growth-factor responsive human AWSM proliferation, and that the oxidase signals in part through activation of the prototypical redox-regulated transcription factor NF-kappaB. PMID- 11880306 TI - Time-dependent reversal of sepsis-induced PMN uptake and lung vascular injury by expression of CD18 antagonist. AB - We determined the time-dependent effects of conditional expression of neutrophil inhibitory factor (NIF), a specific 41-kDa CD18 integrin antagonist, on the time course of NIF expression and lung PMN (polymorphonuclear leukocyte) infiltration and vascular injury in a model of Escherichia coli-induced sepsis in mice. Studies were made in mice transduced with the E-selectin (ES) promoter-NIF construct (using liposomes) in which the NIF cDNA was driven by the inflammation- and endothelial cell-specific ES promoter. We observed time-dependent expression of NIF in pulmonary vascular endothelium that paralleled the ES expression. Expression of both was evident at 1 h after E. coli challenge, peaked at 3-6 h, and returned to basal level within 48 h. We observed that increases in PMN uptake and transalveolar PMN migration induced by E. coli challenge were reversed in a time-dependent manner following NIF expression in mice. NIF expression also prevented the progression of lung vascular injury and edema formation following E. coli challenge. Thus the conditional expression of NIF using the ES promoter can reverse, in a time-dependent manner, lung PMN infiltration and vascular injury induced by gram-negative sepsis. The results support the model that initial engagement of CD18 integrins enables the further recruitment of additional PMN into lung tissues such that PMN continue to sequester and migrate after E. coli challenge. PMID- 11880307 TI - Regulation of surfactant proteins by LPS and proinflammatory cytokines in fetal and newborn lung. AB - Intra-amniotic lipopolysaccharide (LPS) and cytokines may decrease respiratory distress syndrome (RDS) and increase chronic lung disease in the newborn. The aim was to identify the primary inflammatory mediators regulating the expression of surfactant proteins (SP) in explants from immature (22-day-old fetus) and mature (30-day term fetus and 2-day-old newborn) rabbits. In immature lung, interleukin (IL)-1alpha and IL-1beta upregulated the expression of SP-A and SP-B. These effects of IL-1 were diminished, and SP-C mRNA was suppressed additively in the presence of tumor necrosis factor (TNF)-alpha and either LPS or interferon (IFN) gamma. LPS, TNF-alpha, or IFN-gamma had no effect alone. In explants from the term fetus and the newborn, LPS, IL-1alpha, and TNF-alpha additively suppressed the SPs. LPS acutely induced IL-1alpha in alveolar macrophages in mature lung but not in the immature lung. IFN-gamma that generally has low expression in intrauterine infection decreased the age dependence of the other agonists' effects on SPs. The present study serves to explain the variation of the pulmonary outcome after an inflammatory insult. We propose that IL-1 from extrapulmonary sources induces the SPs in premature lung and is responsible for the decreased risk of RDS in intra-amniotic infection. PMID- 11880308 TI - Angiogenic factors and alveolar vasculature: development and alterations by injury in very premature baboons. AB - Proper formation of the pulmonary microvasculature is essential for normal lung development and gas exchange. Lung microvascular development may be disrupted by chronic injury of developing lungs in clinical diseases such as bronchopulmonary dysplasia. We examined microvascular development, angiogenic growth factors, and endothelial cell receptors in a fetal baboon model of chronic lung disease (CLD). In the last third of gestation, the endothelial cell marker platelet endothelial cell adhesion molecule (PECAM)-1 increased 7.5-fold, and capillaries immunostained for PECAM-1 changed from a central location in airspace septa to a subepithelial location. In premature animals delivered at 67% of term and supported with oxygen and ventilation for 14 days, PECAM-1 protein and capillary density did not increase, suggesting failure to expand the capillary network. The capillaries of the CLD animals were dysmorphic and not subepithelial. The angiogenic growth factor vascular endothelial growth factor (VEGF) and its receptor fms-like tyrosine kinase receptor (Flt-1) were significantly decreased in CLD. Angiopoietin-1, another angiogenic growth factor, and its receptor tyrosine kinase with immunoglobulin and epidermal growth factor homology domains were not significantly changed. These data suggest that CLD impairs lung microvascular development and that a possible mechanism is disruption of VEGF and Flt-1 expression. PMID- 11880309 TI - Serum SP-D is a marker of lung injury in rats. AB - Pulmonary surfactant protein D (SP-D) is expressed in alveolar type II and bronchiolar epithelial cells and is secreted into alveoli and conducting airways. However, SP-D has also been measured in serum and is increased in patients with acute respiratory distress syndrome, pulmonary fibrosis, and alveolar proteinosis. To demonstrate that SP-D can be measured in rat serum, we instilled rats with keratinocyte growth factor, which produces type II cell hyperplasia and an increase in SP-D in bronchoalveolar lavage fluid (BALF). To evaluate serum SP D as a biomarker of lung injury, we examined several injury models. In rats treated with 1 unit of bleomycin, serum SP-D was elevated on days 3, 7, 14, and 28 after instillation, and SP-D mRNA was increased in focal areas as detected by in situ hybridization. However, there was no increase in whole lung SP-D mRNA when the expression was normalized to whole lung 18S rRNA. After instillation of 2 units of bleomycin, the serum levels of SP-D were higher, and SP-D was also increased in BALF and lung homogenates. In another model of subacute injury, serum SP-D was increased in rats treated with paraquat plus oxygen. Finally to evaluate acute lung injury, we instilled rats with HCl; SP-D was increased at 4 h after instillation. Our data indicate that serum SP-D may be a useful indicator of lung injury and type II cell hyperplasia in rats. PMID- 11880310 TI - Distribution of intranasal instillations in mice: effects of volume, time, body position, and anesthesia. AB - Intranasal instillation techniques are used to deliver various substances to the upper and lower respiratory tract (URT and LRT) in mice. Here, we quantify the relative distribution achieved with intranasal delivery of a nonabsorbable tracer, (99m)Tc-labeled sulfide-colloid. Relative distribution was determined by killing mice after instillation and quantifying the radioactivity in dissected tissues using gamma scintigraphy. A significant effect of delivery volume on relative distribution was observed when animals were killed 5 min after instillation delivered under gas anesthesia. With a delivery volume of 5 microl, no radiation was detected in the LRT; this increased to a maximum of 55.7 +/- 2.5% distribution to the LRT when 50 microl were delivered. The majority of radiation not detected in the LRT was found in the URT. Over the course of the following 1 h, radiation in the LRT remained constant, while that in the URT decreased and appeared in the gastrointestinal tract. Instillation of 25 microl into anesthetized mice resulted in 30.1 +/- 6.9% distribution to the LRT, while only 5.3 +/- 1.5% (P < 0.05) of the same volume was detected in the LRT of awake mice. Varying the body position of mice did not affect relative distribution. When using intranasal instillation, the relative distribution between the URT and LRT and the gastrointestinal tract is heavily influenced by delivery volume and level of anesthesia. PMID- 11880311 TI - Regulation of ornithine decarboxylase and polyamine import by hypoxia in pulmonary artery endothelial cells. AB - In rat lung and cultured lung vascular cells, hypoxia decreases ornithine decarboxylase (ODC) activity and increases polyamine import. In this study, we used rat cultured pulmonary artery endothelial cells to explore the mechanism of hypoxia-induced reduction in ODC activity and determined whether this event was functionally related to the increase in polyamine import. Two strategies known to suppress proteasome-mediated ODC degradation, lactacystin treatment and use of cells expressing a truncated ODC incapable of interacting with the proteasome, prevented the hypoxia-induced decrease in ODC activity. Interestingly, though, cellular abundance of the 24-kDa antizyme, a known physiological accelerator of ODC degradation, was not increased by hypoxia. These observations suggest that an antizyme-independent ODC degradation pathway contributes to hypoxia-induced reductions of ODC activity. When reductions in ODC activity in hypoxia were prevented by the proteasome inhibitor strategies, hypoxia failed to increase polyamine transport. The induction of polyamine transport in hypoxic pulmonary artery endothelial cells thus seems to require decreased ODC activity as an initiating event. PMID- 11880312 TI - IL-13 and IL-4 cause eotaxin release in human airway smooth muscle cells: a role for ERK. AB - Human airway smooth muscle (HASM) cells express interleukin (IL)-13 and IL-4 receptors and respond to these cytokines with signal transducer and activator of transcription-6 and extracellular signal-regulated kinase (ERK) activation. The purpose of this study was to determine whether IL-13 and/or IL-4 influence eotaxin release in HASM cells and whether the ERK mitogen-activated protein (MAP) kinase pathway is involved in these events. Eotaxin release into HASM cell supernatants was assayed by ELISA, and eotaxin mRNA expression was determined by Northern blot analysis. Pretreatment with either IL-13 or IL-4 resulted in a concentration- and time-dependent release of eotaxin, although IL-4 was more effective. Eotaxin release was approximately twice baseline after treatment with 50 ng/ml IL-13 or IL-4 (P < 0.001). IL-13 and IL-4 also acted synergistically with tumor necrosis factor (TNF)-alpha to induce eotaxin release: TNF-alpha alone (10 ng/ml for 24 h) resulted in an approximately fourfold increase in eotaxin release, whereas TNF-alpha in combination with IL-13 or IL-4 resulted in 10- or 20-fold increases (P < 0.05). Similar results were obtained for eotaxin mRNA expression. Pretreatment with either U-0126 (10 microM) or PD-98059 (30 microM), both inhibitors of MAP/ERK kinase, the enzyme upstream of ERK, inhibited IL-13- or IL-4-induced eotaxin release (P < 0.05). U-0126 also inhibited IL-13, and TNF alpha induced mRNA expression. Our results indicate that IL-13 and IL-4 cause eotaxin release in HASM cells through a mechanism that, in part, involves ERK activation and suggest that the smooth muscle may be an important source of chemokines leading to eosinophil recruitment in asthma. PMID- 11880313 TI - Phosphatidylinositol 3-kinase but not tuberin is required for PDGF-induced cell migration. AB - The loss of function of the tumor suppressor gene TSC2 and its protein product tuberin promotes the development of benign lesions by stimulating cell growth, although the role of tuberin in regulating cell migration and metastasis has not been characterized. In addition, the role of phosphatidylinositol 3-kinase (PI 3 kinase), an important signaling event regulating cell migration, in modulating tuberin-deficient cell motility remains unknown. Using a tuberin-deficient rat smooth muscle cell line, ELT3, we demonstrate that platelet-derived growth factor (PDGF) stimulates cell migration by 3.2-fold, whereas vascular endothelial growth factor (VEGF), transforming growth factor (TGF)-alpha, and basic fibroblast growth factor (bFGF) increase migration by 2.1-, 2.1-, and 2.6-fold, respectively. Basal and PDGF-induced migration in tuberin-deficient ELT3, ELT4, and ERC15 cells was not significantly different from that of tuberin-positive transformed rat kidney epithelial 2, airway smooth muscle, and pulmonary arterial vascular smooth muscle cells. Expression of tuberin in tuberin-deficient ELT3 cells also had little effect on cell migration. In parallel experiments, the role of PI 3-kinase activation in ELT3 cell migration was investigated. LY-294002, a PI 3-kinase inhibitor, decreased PDGF-induced migration in a concentration dependent manner with an IC(50) of approximately 5 microM. LY-294002 also abrogated ELT3 cell migration stimulated by bFGF and TGF-alpha but not by VEGF and phorbol 12-myristate 13-acetate. Furthermore, transient expression of constitutively active PI 3-kinase (p110*) was sufficient to induce ELT3 cell migration. However, the migration induced by p110* was less than that induced by growth factors, suggesting other signaling pathways are also critically important in modulating growth factor-induced cell migration. These data suggest that PI 3 kinase is required for growth factor-induced cell migration and loss of tuberin appears to have little effect on cell migration. PMID- 11880314 TI - Protective role of retinoic acid from antiproliferative action of TNF-alpha on lung epithelial cells. AB - Tumor necrosis factor (TNF)-alpha is a key molecule in lung inflammation. We have established the insulin-like growth factor binding protein 2 (IGFBP-2) as a marker associated with the growth arrest of lung alveolar epithelial cells (AEC). Here, we studied the effects of TNF-alpha on AEC proliferation and the putative protective role of retinoic acid (RA). We documented an antiproliferative action of TNF-alpha that was reversible only at 24 h and then became irreversible with induction of apoptosis. TNF-alpha treatment was associated with a dramatic induction of IGFBP-2. To discover the mechanism of action of IGFBP-2, we further tested the mitogenic potential of IGF-I to counteract TNF-alpha inhibition. Addition of IGF-I to the TNF-alpha containing medium did not stimulate proliferation, whereas des(1-3)IGF-I, an analog of IGF-I that bears low affinity for IGFBPs, was able to restore cell growth. Interestingly, we observed that RA abrogated TNF-alpha-induced growth arrest and that this effect was associated with a dramatic decrease in IGFBP-2 expression. These results suggest a protective role of RA from TNF-alpha antiproliferative action, through mechanisms involving modulation of IGFBP-2 production. PMID- 11880316 TI - New ideas about aldosterone signaling in epithelia. AB - The systemic actions of aldosterone are well documented; however, in comparison, our understanding of the cellular and molecular mechanisms by which aldosterone orchestrates these actions is rudimentary. Aldosterone exerts most of its physiological actions by modifying gene expression. It is now apparent that aldosterone represses almost as many genes as it induces. Several aldosterone sensitive genes, including serum and glucocorticoid-inducible kinase (sgk) and small, monomeric Kirsten Ras GTP-binding protein (Ki-ras) have recently been identified. The molecular mechanisms and elements bestowing corticosteroid sensitivity on these and many other genes are becoming clear. Induction of Ki-Ras and Sgk is necessary and sufficient for some portion of aldosterone action in epithelia. These two signaling factors are components of a converging pathway with phosphatidylinositol 3-kinase positioned between them that enables both stabilizing the epithelial Na(+) channel (ENaC) in the open state as well as increasing the number of ENaC in the apical membrane. This aldosterone-induced signaling pathway contains many potential sites for feedback regulation and cross talk from other cascades and potentially impinges directly on the activity of transport proteins and/or cellular differentiation to modify electrolyte transport. PMID- 11880317 TI - Decreased abundance of collecting duct urea transporters UT-A1 and UT-A3 with ECF volume expansion. AB - Clinical disorders of extracellular fluid (ECF) volume regulation are often associated with changes in plasma urea concentration. To investigate possible renal causes, we measured the relative abundance of the urea transporters UT-A1, UT-A2, and UT-A3 in renal medulla of rats with aldosterone-induced NaCl retention. ECF volume-expanded rats received aldosterone by osmotic minipump plus a diet containing a high level of NaCl. Control rats received the same infusion of aldosterone plus a virtually NaCl-free diet, which prevented ECF volume expansion. Preliminary measurements demonstrated transient positive Na and water balance, decreased serum urea concentration, and increased urea clearance, but no change in creatinine clearance. Immunoblotting of homogenates from inner medulla showed a marked decrease in the abundance of the collecting duct urea transporters UT-A1 and UT-A3. There were no differences in the abundance of UT A2, aquaporin (AQP)-2, AQP-3, or AQP-4 in ECF volume-expanded rats vs. controls. Time course experiments demonstrated that changes in UT-A1 abundance paralleled the fall in serum urea concentration after the switch from a low-NaCl to a high NaCl diet, whereas the fall in UT-A3 abundance was delayed. Candesartan administration markedly decreased the abundance of UT-A1 and UT-A3 in the renal inner medulla, which is consistent with a role for the angiotensin II type 1 receptor in urea transport regulation. The results support the view that ECF related changes in serum urea concentration are mediated, at least in part, through altered urea transporter abundance. PMID- 11880318 TI - Podocyte slit-diaphragm protein nephrin is linked to the actin cytoskeleton. AB - Nephrin is an Ig-like transmembrane protein. It is a major component of the podocyte slit diaphragm and is essential for maintaining normal glomerular permeability. CD2-associated protein (CD2AP) is also necessary for normal glomerular permeability and is a putative nephrin adapter molecule. Here, we document that nephrin and CD2AP are linked to the actin cytoskeleton. As detected by Western blot analysis, nephrin and CD2AP were both insoluble when cell membranes from normal rat glomeruli were extracted with 0.5% Triton X-100 (TX 100) at 4 degrees C in the presence of divalent cations, but they were solubilized when the extraction included potassium iodide (KI) to depolymerize F actin. In addition, a small fraction of the solubilized nephrin and CD2AP was recovered in the low-density fractions of OptiPrep flotation gradients, which indicates that a portion of nephrin, possibly associated with CD2AP, resides in a cholesterol- or sphingolipid-rich region of the plasma membrane. Immunofluorescent staining of unfixed sections of normal rat kidney for nephrin, CD2AP, and F-actin was unaltered by treatment with TX-100 but was greatly diminished by addition of KI. Nephrin staining was slightly reduced by cholesterol depletion with methyl-beta-cyclodextrin in the presence of TX-100 but was nearly absent after addition of KI. These results document that nephrin anchors the slit diaphragm to the actin cytoskeleton, possibly by linkage to CD2AP, and that nephrin traverses a relatively cholesterol-poor region of the podocyte plasma membrane. In addition, a small pool of actin-associated nephrin and CD2AP resides in lipid rafts, possibly in the cholesterol-rich apical region of the podocyte-foot processes. PMID- 11880315 TI - Activation of type II alveolar epithelial cells during acute endotoxemia. AB - Lung injury induced by acute endotoxemia is associated with increased generation of inflammatory mediators such as nitric oxide and eicosanoids, which have been implicated in the pathophysiological process. Although production of these mediators by alveolar macrophages (AM) has been characterized, the response of type II cells is unknown and was assessed in the present studies. Acute endotoxemia caused a rapid (within 1 h) and prolonged (up to 48 h) induction of nitric oxide synthase-2 (NOS-2) in type II cells but a delayed response in AM (12 24 h). In both cell types, this was associated with increased nitric oxide production. Although type II cells, and to a lesser extent AM, constitutively expressed cyclooxygenase-2, acute endotoxemia did not alter this activity. Endotoxin administration had no effect on mitogen-activated protein kinase or protein kinase B-alpha (PKB-alpha) expression. However, increases in phosphoinositide 3-kinase and phospho-PKB-alpha were observed in type II cells. The finding that this was delayed for 12-24 h suggests that these proteins do not play a significant role in the regulation of NOS-2 in this model. After endotoxin administration to rats, a rapid (within 1-2 h) activation of nuclear factor kappaB was observed. This response was transient in type II cells but was sustained in AM. Interferon regulatory factor-1 (IRF-1) was also activated rapidly in type II cells. In contrast, IRF-1 activation was delayed in AM. These data demonstrate that type II cells, like AM, are highly responsive during acute endotoxemia and may contribute to pulmonary inflammation. PMID- 11880319 TI - Angiotensin II stimulates cyclooxygenase-2 mRNA expression in renal tissue from rats with kidney failure. AB - We have shown increased cyclooxygenase-2 (COX-2) expression in rats with kidney failure. Increased angiotensin II concentration, hypertension, and renal mass reduction have been described during development of kidney failure. Thus we explored each of these mechanisms, because any one of them could be responsible for COX-2 induction. Kidney failure increased systolic blood pressure from 104 +/ 5 to 138 +/- 2 mmHg, urinary PGE(2) from 74 +/- 17 to 185 +/- 25 ng/24 h, and COX-2 expression from 0.06 +/- 0.04 to 0.17 +/- 0.03 arbitraty units (AU). Treatment of the rats with ramipril or losartan prevented the increase in blood pressure, urinary PGE(2), and COX-2 expression in the rats with kidney failure. Infusion of angiotensin II increased blood pressure from 101 +/- 6 to 132 +/- 6 mm Hg, urinary PGE(2) excretion from 62 +/- 15 to 155 +/- 17 ng/24 h, and COX-2 expression from 0.23 +/- 0.01 to 1.6 +/- 0.3 AU. When the angiotensin II-infused rats were treated with nitrendipine, blood pressure decreased from 132 +/- 6 to 115 +/- 2 mm Hg, and urinary PGE(2) excretion decreased from 152 +/- 18 to 97 +/- 12 ng/24 h, whereas COX-2 expression was 1.6 +/- 0.7 and 1.7 +/- 0.5 AU for rats with and without nitrendipine. Blood pressure of the rats with renal pole resection was similar to that in sham rats (97 +/- 7 and 91 +/- 4 mmHg, respectively), whereas COX-2 expression was increased in rats with renal pole resection, from 0.06 +/- 0.04 to 0.12 +/- 0.03 AU. We suggest that in kidney failure, the increase in angiotensin II concentration regulates COX-2 expression, thereby increasing prostaglandin synthesis, which contributes to the development of kidney failure. PMID- 11880320 TI - Functional differences between flounder and rat thiazide-sensitive Na-Cl cotransporter. AB - The purpose of the present study was to determine the major functional, pharmacological, and regulatory properties of the flounder thiazide-sensitive Na Cl cotransporter (flTSC) to make a direct comparison with our recent characterization of the rat TSC (rTSC; Monroy A, Plata C, Hebert SC, and Gamba G. Am J Physiol Renal Physiol 279: F161-F169, 2000). When expressed in Xenopus laevis oocytes, flTSC exhibits lower affinity for Na(+) than for Cl(-), with apparent Michaelis-Menten constant (K(m)) values of 58.2 +/- 7.1 and 22.1 +/- 4.2 mM, respectively. These K(m) values are significantly higher than those observed in rTSC. The Na(+) and Cl(-) affinities decreased when the concentration of the counterion was lowered, suggesting that the binding of one ion increases the affinity of the transporter for the other. The effect of several thiazides on flTSC function was biphasic. Low concentrations of thiazides (10(-9) to 10(-7) M) resulted in activation of the cotransporter, whereas higher concentrations (10( 6) to 10(-4) M) were inhibitory. In rTSC, this biphasic effect was observed only with chlorthalidone. The affinity for thiazides in flTSC was lower than in rTSC, but the affinity in flTSC was not affected by the Na(+) or the Cl(-) concentration in the uptake medium. In addition to thiazides, flTSC and rTSC were inhibited by Hg(2+), with an apparent higher affinity for rTSC. Finally, flTSC function was decreased by activation of protein kinase C with phorbol esters and by hypertonicity. In summary, we have found significant regulatory, kinetic, and pharmacological differences between flTSC and rTSC orthologues. PMID- 11880321 TI - Isolation of mouse THP gene promoter and demonstration of its kidney-specific activity in transgenic mice. AB - Tamm-Horsfall protein (THP), the most abundant urinary protein synthesized by the kidney epithelial cells, is believed to play important and diverse roles in the urinary system, including renal water balance, immunosuppression, urinary stone formation, and inhibition of bacterial adhesion. In the present study, we describe the isolation of a 9.3-kb, 5'-region of the mouse THP gene and show the highly conserved nature of its proximal 589-bp, 5'-flanking sequence with that in rats, cattle, and humans. We also demonstrate using the transgenic mouse approach that a 3.0-kb, proximal 5'-flanking sequence is sufficient to drive the kidney specific expression of a heterologous reporter gene. Within the kidney, transgene expression was confined to the renal tubules that endogenously expressed the THP protein, which suggests specific transgene activity in the thick ascending limb of the loop of Henle and early distal convoluted tubules. Our results establish the kidney- and nephron-segment-specific expression of the mouse THP gene. The availability of the mouse THP gene promoter that functions in vivo should facilitate additional studies of the molecular mechanisms of kidney-specific gene regulation and should provide new molecular tools for better understanding renal physiology and disease through nephron-specific gene targeting. PMID- 11880322 TI - Expression of PGT in MDCK cell monolayers: polarized apical localization and induction of active PG transport. AB - The PG transporter (PGT) is expressed in subapical vesicles in the kidney collecting duct. To gain insight into the possible function of the PGT in this tubule segment, we tagged rat PGT with green fluorescent protein at the COOH terminus and generated stable PGT-expressing Madin-Darby canine kidney cell lines. When grown on permeable filters, green fluorescent protein-PGT was expressed predominantly at the apical membrane. Although the basal-to-apical transepithelial flux of [(3)H]PGE(2) was little changed by PGT expression, the apical-to-basolateral flux was increased 100-fold compared with wild-type cells. Analysis of driving forces revealed that this flux represents PGT-mediated active transepithelial PGE(2) transport. We propose that endogenous PGT is exocytically inserted into the collecting duct apical membrane, where it could control the concentration of luminal PGs. PMID- 11880323 TI - Role of internalization in AT(1A) receptor function in proximal tubule epithelium. AB - Angiotensin II (ANG II), acting through angiotensin type I (AT(1)) receptors on apical and basolateral surfaces of proximal tubule epithelial cells, increases sodium reabsorption in proximal tubules. Apical and basolateral receptors internalize after exposure to ANG II, but the role of internalization in receptor signaling and transport is not well defined. To determine the role of receptor internalization in ANG II-mediated receptor signaling and sodium transport, we stably expressed full-length and truncated AT(1A) receptors in opossum kidney cells. After stimulation with ANG II, wild-type receptors on apical and basolateral surfaces rapidly internalized, inhibited adenylate cyclase, and increased transcellular sodium transport. Truncation of the cytoplasmic tail of the AT(1A) receptor (TL314) resulted in receptors that were expressed on apical and basolateral surfaces but did not internalize, inhibit adenylate cyclase, or increase sodium transport. Because the cytoplasmic tail contains putative G protein coupling sites, mutant receptors that leave G protein interaction sites intact were designed. Cells expressing the truncation (TK333) or deletion (Del 315-329) also failed to internalize. When ANG II was added to basolateral surfaces of TK333 or Del 315-329, adenylate cyclase activity was inhibited and sodium transport was increased. In contrast, apical addition of ANG II was not associated with decreases in adenylate cyclase or increases in sodium transport. In conclusion, internalization pathways are important for AT(1A) receptor function in polarized proximal tubule epithelial cells. Apical AT(1A) receptors internalize before they interact with G proteins and signal cAMP. In contrast, basolateral AT(1A) receptors interact with G proteins and signal cAMP without internalizing. PMID- 11880324 TI - Structure and characterization of the mouse UT-A gene (Slc14a2). AB - The movement of urea across plasma membranes is modulated by facilitated urea transporter proteins. These proteins are the products of two closely related genes, termed UT-A (Slc14a2) and UT-B (Slc14a1). By genomic library screening and P1 artificial chromosome "shotgun" sequencing, we have determined the structure of the mouse UT-A gene. The gene is >300 kb in length, contains 24 exons, and has 2 distinct promoters. Flanking the 5'-region of the gene is the UT-Aalpha promoter that regulates transcription of UT-A1 and UT-A3. The second promoter, termed UT-Abeta, is present in intron 13 and regulates transcription of UT-A2. cAMP agonists (100 microM dibutryl cAMP, 25 microM forskolin, 0.5 mM IBMX) increased the activity of a 2.2-kb UT-Aalpha promoter construct 6.2-fold [from 0.026 +/- 0.003 to 0.160 +/- 0.004, relative light units (RLU)/microg protein] and a 2.4-kb UT-Abeta promoter construct 9.5-fold (from 0.020 +/- 0.002 to 0.190 +/- 0.043 RLU/microg protein) above that in untreated controls. Interestingly, only the UT-Abeta promoter contained consensus sequences for CREs and deletion of these elements abolished cAMP sensitivity. Increasing the tonicity of culture medium from 300 to 600 mosmol/kg H(2)O with NaCl caused a significant increase (from 0.060 +/- 0.004 to 0.095 +/- 0.010 RLU/microg protein) in UT-Aalpha promoter activity but had no effect on the UT-Abeta promoter. A tonicity responsive enhancer was identified in UT-Aalpha and is suggested to be responsible for mediating this effect. Levels of UT-A2 and UT-A3 mRNA were increased in thirsted mice compared with control animals, indicating that the activities of both promoters are likely to be elevated during prolonged antidiuresis. PMID- 11880325 TI - Upregulation of type I collagen by TGF-beta in mesangial cells is blocked by PPARgamma activation. AB - We found that peroxisome proliferator-activated receptor-gamma (PPARgamma) mRNA was reduced by 77% in glomeruli of diabetic mice. Because mesangial cells play an important role in diabetic nephropathy, we examined regulation of type I collagen expression by PPARgamma and transforming growth factor-beta(1) (TGF-beta(1)) in mouse mesangial cells in the presence of 6 and 25 mM glucose. Mesangial cells contained functionally active PPARgamma. Exposure to 25 mM glucose resulted in reduced PPARgamma expression and transcriptional activity, accompanied by increased type I collagen expression. Restoration of PPARgamma activity to normal levels in cells cultured in 25 mM glucose, by transfection with a PPARgamma expression construct and treatment with the PPARgamma agonist troglitazone, returned type I collagen levels toward normal values. Activation of PPARgamma by troglitazone also decreased type I collagen mRNA and blocked TGF-beta(1)-mediated upregulation of type I collagen mRNA and protein. Moreover, PPARgamma activation suppressed basal and activated TGF-beta(1) responses in mesangial cells. This action was blocked by transfection of cells with a dominant-negative PPARgamma construct. In summary, PPARgamma suppresses the increased type I collagen mRNA and protein expression mediated by TGF-beta(1) in mesangial cells. PMID- 11880326 TI - Expression of isoforms of the Na(+)/H(+) exchanger in M-1 mouse cortical collecting duct cells. AB - Na(+)/H(+) exchanger (NHE) proteins perform a variety of functions in the kidney and are differentially distributed among nephron segments. The purpose of this study was to identify NHE isoforms in murine M-1 cells as a model of cortical collecting duct principal cells. It was found that mRNAs corresponding to NHE1, NHE2, and NHE4 are expressed in M-1 cells. NHE-dependent regulation of intracellular pH (pH(i)) was investigated in the absence of extracellular HCO. Application of a 20 mM NH(4)Cl pulse resulted in a reversible intracellular acidification from which recovery was partially inhibited by application of 1 mM amiloride to either the apical or the basolateral membranes and was abolished when amiloride was applied to both sides of the monolayers, which suggests that NHEs are expressed in both the apical and the basolateral cell membranes of M-1 cells. The purinergic agonists ATP and benzoylbenzoyl-ATP caused a reduction of pH(i) when applied to the apical membrane, which suggests pH(i) may be influenced by extracellular nucleotides in the luminal fluid of the cortical collecting duct. PMID- 11880327 TI - Basolateral membrane Cl(-)-, Na(+)-, and K(+)-coupled base transport mechanisms in rat MTALH. AB - Mechanisms involved in basolateral HCO transport were examined in the in vitro microperfused rat medullary thick ascending limb of Henle (MTALH) by microfluorometric monitoring of cell pH. Removing peritubular Cl(-) induced a cellular alkalinization that was inhibited in the presence of peritubular 4,4' diisothiocyanostilbene-2,2'-disulfonic acid (DIDS) and blunted in the absence of external CO(2)/HCO. The alkalinization elicited by removing peritubular Cl(-) persisted in the bilateral absence of Na(+), together with a voltage clamp. When studied in Cl(-)-free solutions, lowering peritubular pH induced a base efflux that was inhibited by peritubular DIDS or by the absence of external CO(2)/HCO. Removing peritubular Na(+) elicited a cellular acidification that was accounted for by stimulation of a DIDS- and ethylisopropylamiloride (EIPA)-insensitive Na(+)-HCO cotransport and inhibition of a basolateral Na(+)/H(+) exchange. Increasing bath K(+) induced an intracellular alkalinization that was inhibited in the absence of external CO(2)/HCO. At 2 mM, peritubular Ba(2+), which inhibits the K(+)-Cl(-) cotransport, did not induce any change in transepithelial voltage but elicited a cellular alkalinization and inhibited K(+)-induced cellular alkalinization, consistent with the presence of a basolateral, electroneutral Ba(2+)-sensitive K(+)-Cl(-) cotransport that may operate as a K(+)-HCO cotransport. This cotransport was inhibited in the peritubular presence of furosemide, [(dihydroindenyl)oxy]alkanoic acid, 5-nitro-2-(3 phenylpropylamino)benzoate, or DIDS. At least three distinct basolateral HCO transport mechanisms are functional under physiological conditions: electroneutral Cl(-)/HCO exchange, DIDS- and EIPA-insensitive Na(+)-HCO cotransport, and Ba(2+)-sensitive electroneutral K(+)-Cl(-)(HCO) cotransport. PMID- 11880328 TI - Aldosterone interaction with epidermal growth factor receptor signaling in MDCK cells. AB - Epidermal growth factor (EGF) regulates cell proliferation, differentiation, and ion transport by using extracellular signal-regulated kinase (ERK)1/2 as a downstream signal. Furthermore, the EGF-receptor (EGF-R) is involved in signaling by G protein-coupled receptors, growth hormone, and cytokines by means of transactivation. It has been suggested that steroids interact with peptide hormones, in part, by rapid, potentially nongenomic, mechanisms. Previously, we have shown that aldosterone modulates Na(+)/H(+) exchange in Madin-Darby canine kidney (MDCK) cells by means of ERK1/2 in a way similar to growth factors. Here, we tested the hypothesis that aldosterone uses the EGF-R as a heterologous signal transducer in MDCK cells. Nanomolar concentrations of aldosterone induce a rapid increase in ERK1/2 phosphorylation, cellular Ca(2+) concentration, and Na(+)/H(+) exchange activity similar to increases induced by EGF. Furthermore, aldosterone induced a rapid increase in EGF-R-Tyr phosphorylation, and inhibition of EGF-R kinase abolished aldosterone-induced signaling. Inhibition of ERK1/2 phosphorylation reduced the Ca(2+) response, whereas prevention of Ca(2+) influx did not abolish ERK1/2 phosphorylation. Our data show that aldosterone uses the EGF-R-ERK1/2 signaling cascade to elicit its rapid effects in MDCK cells. PMID- 11880329 TI - Role of the cytoskeleton in mediating effect of vasopressin and herbimycin A on secretory K channels in CCD. AB - We have previously demonstrated that inhibiting protein tyrosine kinase (PTK) and stimulating protein kinase A (PKA) increase the activity of the small-conductance K (SK) channel in the cortical collecting duct (CCD) of rat kidneys (Cassola AC, Giebisch G, and Wang WH. Am J Physiol Renal Fluid Electrolyte Physiol 264: F502 F509, 1993; Wang WH, Lerea KM, Chan M, and Giebisch G. Am J Physiol Renal Physiol 278: F165-F171, 2000). In the present study, we used the patch-clamp technique to study the role of the cytoskeleton in mediating the effect of herbimycin A, an inhibitor of PTK, and vasopressin on the SK channels in the CCD. The addition of colchicine, an inhibitor of microtubule assembly, or taxol, an agent that blocks microtubule reconstruction, had no significant effect on channel activity. However, colchicine and taxol treatment completely abolished the stimulatory effect of herbimycin A on the SK channels in the CCD. Removal of the microtubule inhibitors restored the stimulatory effect of herbimycin A. In contrast, treatment of the tubules with either taxol or colchicine did not block the stimulatory effect of vasopressin on the SK channels. Moreover, the effect of herbimycin A on the SK channels was also absent in the CCDs treated with either cytochalasin D or phalloidin. In contrast, the stimulatory effect of vasopressin was still observed in the tubules treated with phalloidin. However, cytochalasin D treatment abolished the effect of vasopressin on the SK channels. Finally, the effects of vasopressin and herbimycin A are additive because inhibiting PTK can still increase the channel activity in CCD that has been challenged by vasopressin. We conclude that an intact cytoskeleton is required for the effect on the SK channels of inhibiting PTK and that the SK channels that are activated by inhibiting PTK were differently regulated from those stimulated by vasopressin. PMID- 11880330 TI - Identification of functionally important sites in the first intracellular loop of the NaPi-IIa cotransporter. AB - Intrasequence comparison of the type IIa Na(+)-P(i) cotransport protein revealed two regions with high similarity in the first intracellular (ICL-1) and third extracellular (ECL-3) loops. Because the ECL-3 loop contains functionally important sites that have been identified by cysteine scanning, we applied this method to corresponding sites in the ICL-1 loop. The accessibility of novel cysteines by methanethiosulfonate reagents was assayed electrophysiologically. Mutants N199C and V202C were fully inhibited after methanethiosulfonate ethylammonium exposure, whereas other mutants showed marginal reductions in cotransport function. None showed significant functional loss after exposure to impermeant methanethiosulfonate ethyltrimethylammonium, which suggested a sidedness of Cys modification. Compared with the wild-type (WT), mutant A203C showed altered Na(+) leak kinetics, whereas N199C exhibited decreased apparent substrate affinities. To delineate the role of residue N199 in conferring substrate affinity, other mutations at this site were made. Only two mutants yielded significant (32)P(i) uptake and inward P(i)-induced currents with decreased P(i) affinity; for the others, P(i) application suppressed only the Na(+) leak. We suggest that ICL-1 and ECL-3 sites contribute to the transport pathway and that site N199 is implicated in defining the transport mode. PMID- 11880331 TI - Role of tyrosine kinase and p44/42 MAPK in D(2)-like receptor-mediated stimulation of Na(+), K(+)-ATPase in kidney. AB - Our laboratory has shown that dopamine D(2)-like receptor activation causes stimulation of Na(+), K(+)-ATPase (NKA) activity in the proximal tubules of the rat kidney. The present study was designed to investigate the cellular signaling mechanisms mediating this response to D(2)-like receptor activation. We measured the stimulation of NKA activity by bromocriptine (D(2)-like receptor agonist) in the absence and presence of PD-98059 [p44/42 mitogen-activated protein kinase (MAPK) kinase inhibitor] and genistein (tyrosine kinase inhibitor) in renal proximal tubules. Both agents inhibited bromocriptine-mediated stimulation of NKA, suggesting the involvement of p44/42 MAPK and tyrosine kinase in this response. Additionally, we found that bromocriptine increased the phosphorylation of p44/42 MAPK in the proximal tubules, which was blocked by PD-98059 and genistein. These results show that D(2)-like receptor activation causes stimulation of NKA activity by means of a tyrosine kinase-p44/42 MAPK pathway in the proximal tubules of the kidney. PMID- 11880332 TI - Clusterin protects renal tubular epithelial cells from gentamicin-mediated cytotoxicity. AB - Clusterin is a heterodimeric secreted glycoprotein that is upregulated after acute renal injury. In aminoglycoside nephrotoxicity, clusterin is induced in the tubular epithelium and increased levels are found in the urine. In this study, we developed an in vitro model of gentamicin-induced cytotoxicity in renal proximal tubule cells and tested whether clusterin protected these cells from injury. LLC PK(1) cells were incubated with varying concentrations of gentamicin in serum free media, and cytotoxicity was quantified by lactate dehydrogenase release and confirmed by vital dye exclusion. A dose-dependent increase in cytotoxicity occurred with gentamicin concentrations up to 27 mg/ml. Clusterin decreased cytotoxicity in a dose- and time-dependent manner at 6, 12, and 24 h, whereas albumin, used as a control protein, had no effect. In contrast to the aminoglycoside model, when cells were injured by depletion of ATP, clusterin had only a minimally protective effect. LLC-PK(1) cells did not express megalin, a receptor that can mediate the uptake of both clusterin and aminoglycosides into proximal tubule cells. Uptake of gentamicin into LLC-PK(1) cells was observed despite the absence of megalin. In conclusion, clusterin specifically protects against gentamicin-induced renal tubular cell injury by a megalin-independent mechanism. PMID- 11880334 TI - Hyperosmotic mannitol activates basolateral NHE in proximal tubule from P glycoprotein null mice. AB - Using the pH-sensitive fluorescent dye 2',7'-bis(carboxyethyl)-5(6) carboxyfluorescein acetoxymethyl ester, we examined the effects of hyperosmotic mannitol on basolateral Na(+)/H(+) exchange (NHE) activity in isolated nonperfused proximal tubule S2 segments from mice lacking both the mdr1a and mdr1b genes (KO) and wild-type mice (WT). All experiments were performed in CO(2)/HCO-free HEPES solutions. Osmolality of the peritubular solution was raised from 300 to 500 mosmol/kgH(2)O by the addition of mannitol. NHE activity was assessed by Na(+)-dependent acid extrusion rates (J(H)) after an acid load with NH(4)Cl prepulse. Under isosmotic conditions, J(H) values at a wide intracellular pH (pH(i)) range of 6.20-6.90 were not different between the two groups. In WT mice, hyperosmotic mannitol had no effect on J(H) at the wide pH(i) range. In contrast, in KO mice, hyperosmotic mannitol increased J(H) at a pH(i) range of 6.20-6.45 and shifted the J(H)-pH(i) relationship by 0.15 pH units in the alkaline direction. In KO mice, hyperosmotic mannitol caused an increase in maximal velocity without changing the Michaelis-Menten constant for peritubular Na(+). Exposure of cells from WT mice to the hyperosmotic mannitol solution including the P-gp inhibitor cyclosporin A increased J(H) (at pH(i) 6.30) to an extent similar to that in cells from KO mice exposed to hyperosmotic mannitol alone. In KO mice, staurosporine and calphostin C inhibited the hyperosmotic mannitol-induced increase in J(H). The stimulatory effect of hyperosmotic mannitol on J(H) was mimicked by addition to the isosmotic control solution, including phorbol 12-myristate 13-acetate (PMA; the PKC activator). In WT mice, hyperosmotic mannitol with PMA increased J(H). We conclude that, in the absence of P-gp activity, hyperosmotic mannitol activates basolateral NHE via protein kinase C, whereas in the presence of P-gp activity, it does not. PMID- 11880333 TI - Rottlerin inhibits tonicity-dependent expression and action of TonEBP in a PKCdelta-independent fashion. AB - Novel protein kinase C (PKC) isoforms PKCdelta and PKCepsilon have recently been implicated in signaling by hypertonic stress. We investigated the role of the putative PKCdelta inhibitor rottlerin on tonicity-dependent gene regulation. In the renal medullary mIMCD3 cell line, rottlerin blocked tonicity-dependent transcription of a tonicity enhancer (TonE)-driven luciferase reporter gene, as well as tonicity-dependent transcription of the physiological tonicity effector gene aldose reductase, but not urea-dependent transcription. Consistent with these data, rottlerin inhibited tonicity-dependent expression of TonE binding protein (TonEBP) at the mRNA and protein levels. Another inhibitor of both novel and conventional PKC isoforms, GF-109203X, suppressed TonEBP-dependent transcription but failed to influence tonicity-inducible TonEBP expression. Global PKC downregulation with protracted phorbol ester treatment, however, failed to influence tonicity-dependent signaling, arguing against a PKCdelta dependent mechanism of rottlerin action in this model. In addition, hypertonic stress failed to induce phosphorylation of PKCdelta. Furthermore, in a PC-12 cell model with a comparable degree of tonicity-dependent transcription, constitutive overexpression of dominant negative-acting PKCdelta or PKCepsilon effectively decreased tonicity signaling to extracellular signal-regulated kinase activation, as expected, but failed to influence TonE-dependent transcription. TonE-dependent transcription, however, remained rottlerin sensitive in this PC-12 cell model. In the aggregate, these data indicate that rottlerin dramatically inhibits tonicity dependent TonEBP expression and TonE-dependent transcription but, despite its reputed mode of action, does so through a PKCdelta-independent pathway. PMID- 11880335 TI - Acute hypertension provokes internalization of proximal tubule NHE3 without inhibition of transport activity. AB - Acute hypertension rapidly decreases proximal tubule (PT) Na(+) reabsorption, facilitated by a redistribution of PT Na(+)/H(+) exchangers (NHE3) out of the apical brush border, increasing NaCl at the macula densa, the signal for autoregulation of renal blood flow and GFR. This study aimed to determine whether NHE3 activity per transporter decreases during acute hypertension and the time dependence of the response. Blood pressure was elevated by 50-60 mmHg in male Sprague-Dawley rats for 5 or 30 min by constricting arteries. Renal cortical membranes were fractionated by density gradient centrifugation. NHE3 transport activity was assayed as the rate of appearance of acridine orange (AO) from AO loaded vesicles in response to an inwardly directed Na(+) gradient. After 5-min hypertension, 20% of total NHE3 protein, assayed by immunoblot, redistributed from low-density apical membranes to middensity membranes enriched in intermicrovillar cleft markers; by 30 min, a similar percentage shifted to heavier density membranes containing markers of endosomes. NHE3 activity shifted to higher density membranes along with NHE3 protein, that is, no change in activity/transporter during acute hypertension. Confocal analysis of NHE3 distribution also verified removal from apical microvilli and appearance in subapical vesicles. We conclude that the decrease in renal PT Na(+) transport during acute hypertension is mediated by removal of transport-competent NHE3 from the apical brush border to subapical and internal reserves. PMID- 11880336 TI - Retinoid receptor-specific agonists alleviate experimental glomerulonephritis. AB - Retinoids are potent antiproliferative and anti-inflammatory compounds. We previously demonstrated that the natural pan-agonists all-trans retinoic acid (RA) and 13-cis RA efficiently preserve renal structure and function in rat mesangioproliferative glomerulonephritis. We examine effects of synthetic retinoid receptor-specific agonists 1) to identify common and receptor subtype specific pathways in this model and 2) to characterize effects of retinoids on the renal endothelin (ET) system. Vehicle-injected control rats were compared with rats treated with daily subcutaneous injections of agonists specific for retinoid A (Ro-137410) and retinoid X (Ro-257386) receptors and the complex anti activator protein-1 active retinoid BMS-453 7 days after induction of anti-Thy1.1 nephritis (n = 7-9/group). The different retinoids lowered glomerular ET-1 and ET type A and B receptor gene expression in control and nephritic rats with comparable efficacy. Reduction of glomerular c-Fos and GATA-2 mRNA expression levels suggests downregulation of transcription factors required for ET expression. The different retinoids were similar in their action on the glomerular capillary occlusion score, number of total glomerular cells, and glomerular infiltrating macrophage count. They differed in their ability to normalize blood pressure (Ro-257386 > BMS-453 > arotinoid), albuminuria (BMS-453 > Ro-257386 > arotinoid), and creatinine clearance (arotinoid > BMS-453 > Ro 257386). No signs of toxicity were observed. We conclude that all retinoid agonists with different subtype specificity are highly efficient in reducing renal damage and proliferation of mesangial cells. Retinoid X and A receptor specific pathways are apparently involved in the antiproliferative, anti inflammatory, and anti-ET action. Further studies are indicated to define the potential use of retinoid agonists in inflammatory renal disease. PMID- 11880337 TI - Relevance of renal-specific oxidoreductase in tubulogenesis during mammalian nephron development. AB - Renal-specific oxidoreductase (RSOR), an enzyme relevant to diabetic nephropathy, is exclusively expressed in renal tubules. Studies were initiated to determine whether, like other tubule-specific proteins, it selectively modulates tubulogenesis. Northern blot analyses revealed a approximately 1.5-kb transcript, and RSOR expression was detectable in mice embryonic kidneys at day 13, gradually increased by day 17, and extended into neo- and postnatal periods. RSOR mRNA and protein expression was confined to proximal tubules, commencing at gestational day 17 and increasing subsequently, but remained absent in glomeruli and medulla. Treatment with RSOR antisense oligodeoxynucleotide resulted in a dose-dependent dysmorphogenesis of metanephric explants harvested at gestational day 13. The explants were smaller and had expanded mesenchyme, and the population of tubules was markedly decreased. The glomeruli were unaffected, as assessed by mRNA expression of glomerular epithelial protein 1 and reactivity with wheat germ agglutinin. Antisense treatment led to a selective reduction of RSOR mRNA. Immunoprecipitation also indicated a selective translational blockade of RSOR. These findings suggest that RSOR is developmentally regulated, exhibits a distinct spatiotemporal distribution, and probably plays a role in tubulogenesis. PMID- 11880338 TI - Autocrine extracellular purinergic signaling in epithelial cells derived from polycystic kidneys. AB - ATP and its metabolites are potent autocrine agonists that act extracellularly within tissues to affect epithelial function. In polycystic kidneys, renal tubules become dilated and/or encapsulated as cysts, creating abnormal microenvironments for autocrine signaling. Previously, our laboratory has shown that high-nanomolar to micromolar quantities of ATP are released from cell monolayers in vitro and detectable in cyst fluids from microdissected human autosomal dominant polycystic kidney (ADPKD) cysts. Here, we show enhanced ATP release from autosomal recessive polycystic kidney (ARPKD) and ADPKD epithelial cell models. RT-PCR and immunoblotting for P2Y G protein-coupled receptors and P2X purinergic receptor channels show expression of mRNA and/or protein for multiple subtypes from both families. Assays of cytosolic Ca(2+) concentration and secretory Cl(-) transport show P2Y and P2X purinergic receptor-mediated stimulation of Cl(-) secretion via cytosolic Ca(2+)-dependent signaling. Therefore, we hypothesize that autocrine purinergic signaling may augment detrimentally cyst volume expansion in ADPKD or tubule dilation in ARPKD, accelerating disease progression. PMID- 11880339 TI - R8 development in the Drosophila eye: a paradigm for neural selection and differentiation. AB - The Drosophila eye is an outstanding model with which to decipher mechanisms of neural differentiation. Paramount to normal eye development is the organized selection and differentiation of a patterned array of R8 photoreceptors - the founding photoreceptor of each ommatidium that coordinates the incorporation of all other photoreceptors. R8 development is a complex process that requires the integration of transcription factors and signaling pathways, many of which are highly conserved and perform similar functions in other species. This article discusses the developmental control of the four key elements of R8 development: selection, spacing, differentiation and orchestration of later events. New questions that have surfaced because of recent advances in the field are addressed, and the unique characteristics of R8 development are highlighted through comparisons with neural specification in other Drosophila tissues and with ganglion cell development in the mammalian retina. PMID- 11880340 TI - eFGF is required for activation of XmyoD expression in the myogenic cell lineage of Xenopus laevis. AB - This paper addresses the molecular mechanisms that regulate the transcriptional activation of the myogenic regulatory factor XmyoD in the skeletal muscle lineage of Xenopus laevis. Using antisense morpholino oligonucleotide-mediated inhibition, we show that the signalling molecule embryonic fibroblast growth factor (eFGF), which is the amphibian homologue of FGF4, is necessary for the initial activation of XmyoD transcription in myogenic cells. We demonstrate that eFGF can activate the expression of XmyoD in the absence of protein synthesis, indicating that this regulation is direct. Our data suggest that regulation of XmyoD expression may involve a labile transcriptional repressor. In addition, we show that eFGF is itself an immediate early response to activin, a molecule that mimics the endogenous mesoderm-inducing signal. We propose a model for the regulation of XmyoD within the early mesoderm, and discuss the relevance that these findings have for the understanding of myogenic specification in higher vertebrates. PMID- 11880341 TI - Lola regulates midline crossing of CNS axons in Drosophila. AB - The pattern and level of expression of axon guidance proteins must be choreographed with exquisite precision for the nervous system to develop its proper connectivity. Previous work has shown that the transcription factor Lola is required for central nervous system (CNS) axons of Drosophila to extend longitudinally. We show here that Lola is simultaneously required to repel these same longitudinal axons away from the midline, and that it acts, in part, by augmenting the expression both of the midline repellant, Slit, and of its axonal receptor, Robo. Lola is thus the examplar of a class of axon guidance molecules that control axon patterning by coordinating the regulation of multiple, independent guidance genes, ensuring that they are co-expressed at the correct time, place and relative level. PMID- 11880342 TI - Pax6 regulates specification of ventral neurone subtypes in the hindbrain by establishing progenitor domains. AB - Recent studies have shown that generation of different kinds of neurones is controlled by combinatorial actions of homeodomain (HD) proteins expressed in the neuronal progenitors. Pax6 is a HD protein that has previously been shown to be involved in the differentiation of the hindbrain somatic (SM) motoneurones and V1 interneurones in the hindbrain and/or spinal cord. To investigate in greater depth the role of Pax6 in generation of the ventral neurones, we first examined the expression patterns of HD protein genes and subtype-specific neuronal markers in the hindbrain of the Pax6 homozygous mutant rat. We found that Islet2 (SM neurone marker) and En1 (V1 interneurone marker) were transiently expressed in a small number of cells, indicating that Pax6 is not directly required for specification of these neurones. We also observed that domains of all other HD protein genes (Nkx2.2, Nkx6.1, Irx3, Dbx2 and Dbx1) were shifted and their boundaries became blurred. Thus, Pax6 is required for establishment of the progenitor domains of the ventral neurones. Next, we performed Pax6 overexpression experiments by electroporating rat embryos in whole embryo culture. Pax6 overexpression in the wild type decreased expression of Nkx2.2, but ectopically increased expression of Irx3, Dbx1 and Dbx2. Moreover, electroporation of Pax6 into the Pax6 mutant hindbrain rescued the development of Islet2-positive and En1-positive neurones. To know reasons for perturbed progenitor domain formation in Pax6 mutant, we examined expression patterns of Shh signalling molecules and states of cell death and cell proliferation. Shh was similarly expressed in the floor plate of the mutant hindbrain, while the expressions of Ptc1, Gli1 and Gli2 were altered only in the progenitor domains for the motoneurones. The position and number of TUNEL-positive cells were unchanged in the Pax6 mutant. Although the proportion of cells that were BrdU positive slightly increased in the mutant, there was no relationship with specific progenitor domains. Taken together, we conclude that Pax6 regulates specification of the ventral neurone subtypes by establishing the correct progenitor domains. PMID- 11880343 TI - Zeste maintains repression of Ubx transgenes: support for a new model of Polycomb repression. AB - During late embryogenesis, the expression domains of homeotic genes are maintained by two groups of ubiquitously expressed regulators: the Polycomb repressors and the Trithorax activators. It is not known how the activities of the two maintenance systems are initially targeted to the correct genes. Zeste and GAGA are sequence-specific DNA-binding proteins previously shown to be Trithorax group activators of the homeotic gene Ultrabithorax (Ubx). We demonstrate that Zeste and GAGA DNA-binding sites at the proximal promoter are also required to maintain, but not to initiate, repression of UBX: Furthermore, the repression mediated by Zeste DNA-binding site is abolished in zeste null embryos. These data imply that Zeste and probably GAGA mediate Polycomb repression. We present a model in which the dual transcriptional activities of Zeste and GAGA are an essential component of the mechanism that chooses which maintenance system is to be targeted to a given promoter. PMID- 11880344 TI - The role of RBF in developmentally regulated cell proliferation in the eye disc and in Cyclin D/Cdk4 induced cellular growth. AB - During Drosophila eye development, cell proliferation is coordinated with differentiation. Immediately posterior to the morphogenetic furrow, cells enter a synchronous round of S phase called second mitotic wave. We have examined the role of RBF, the Drosophila RB family homolog, in cell cycle progression in the second mitotic wave. RBF-280, a mutant form of RBF that has four putative cdk phosphorylation sites mutated, can no longer be regulated by Cyclin D or Cyclin E. Expression of RBF-280 in the developing eye revealed that RBF-280 does not inhibit G1/S transition in the second mitotic wave, rather it delays the completion of S phase and leads to abnormal eye development. These observations suggest that RB/E2F control the rate of S-phase progression instead of G1/S transition in the second mitotic wave. Characterization of the role of RBF in Cyclin D/Cdk4-mediated cellular growth showed that RBF-280 blocks Cyclin D/Cdk4 induced cellular growth in the proliferating wing disc cells but not in the non dividing eye disc cells. By contrast, RBF-280 does not block activated Ras induced cellular growth. These results suggest that the ability of Cyclin D/Cdk4 to drive growth in the proliferating wing cells is distinct from that in the none dividing eye cells or the ability of activated Ras to induce growth, and that RBF may have a role in regulating growth in the proliferating wing discs. PMID- 11880345 TI - Subdivision of the Drosophila wing imaginal disc by EGFR-mediated signaling. AB - Growth and patterning of the Drosophila wing imaginal disc depends on its subdivision into dorsoventral (DV) compartments and limb (wing) and body wall (notum) primordia. We present evidence that both the DV and wing-notum subdivisions are specified by activation of the Drosophila Epidermal Growth Factor Receptor (EGFR). We show that EGFR signaling is necessary and sufficient to activate apterous (ap) expression, thereby segregating the wing disc into D (ap-ON) and V (ap-OFF) compartments. Similarly, we demonstrate that EGFR signaling directs the expression of Iroquois Complex (Iro-C) genes in prospective notum cells, rendering them distinct from, and immiscible with, neighboring wing cells. However, EGFR signaling acts only early in development to heritably activate ap, whereas it is required persistently during subsequent development to maintain Iro-C gene expression. Hence, as the disc grows, the DV compartment boundary can shift ventrally, beyond the range of the instructive EGFR signal(s), in contrast to the notum-wing boundary, which continues to be defined by EGFR input. PMID- 11880346 TI - Control of growth and patterning of the Drosophila wing imaginal disc by EGFR mediated signaling. AB - The subdivision of the Drosophila wing imaginal disc into dorsoventral (DV) compartments and limb-body wall (wing-notum) primordia depends on Epidermal Growth Factor Receptor (EGFR) signaling, which heritably activates apterous (ap) in D compartment cells and maintains Iroquois Complex (Iro-C) gene expression in prospective notum cells. We examine the source, identity and mode of action of the EGFR ligand(s) that specify these subdivisions. Of the three known ligands for the Drosophila EGFR, only Vein (Vn), but not Spitz or Gurken, is required for wing disc development. We show that Vn activity is required specifically in the dorsoproximal region of the wing disc for ap and Iro-C gene expression. However, ectopic expression of Vn in other locations does not reorganize ap or Iro-C gene expression. Hence, Vn appears to play a permissive rather than an instructive role in organizing the DV and wing-notum segregations, implying the existance of other localized factors that control where Vn-EGFR signaling is effective. After ap is heritably activated, the level of EGFR activity declines in D compartment cells as they proliferate and move ventrally, away from the source of the instructive ligand. We present evidence that this reduction is necessary for D and V compartment cells to interact along the compartment boundary to induce signals, like Wingless (Wg), which organize the subsequent growth and differentiation of the wing primordium. PMID- 11880347 TI - An adjunct mammary epithelial cell population in parous females: its role in functional adaptation and tissue renewal. AB - Mammary gland biologists have long assumed that differentiated secretory epithelial cells undergo programmed cell death at the end of lactation and that the alveolar compartment is reconstituted from undifferentiated precursor cells in subsequent pregnancies. It is generally agreed that the remodeled gland in a parous animal resembles that of a mature virgin at the morphological level. However, several physiological differences have been noted in comparing the responses of mammary epithelia from nulliparous versus parous females to hormonal stimulation and carcinogenic agents. We present genetic evidence that an involuted mammary gland is fundamentally different from a virgin gland, despite its close morphological resemblance. This difference results from the formation of a new mammary epithelial cell population that originates from differentiating cells during pregnancy. In contrast to the majority of fully committed alveolar cells, this epithelial population does not undergo cell death during involution or remodeling after lactation. We show that these cells can function as alveolar progenitors in subsequent pregnancies and that they can play an important role in functional adaptation in genetically engineered mice, which exhibit a reversion of a lactation-deficient phenotype in multiparous animals. In transplantation studies, this parity-induced epithelial population shows the capacity for self renewal and contributes significantly to the reconstitution of the resulting mammary outgrowth (i.e. ductal morphogenesis and lobulogenesis). We propose that this parity-induced population contributes importantly to the biological differences between the mammary glands of parous and nulliparous females. PMID- 11880348 TI - The developmental expression of vasoactive intestinal peptide (VIP) in cholinergic sympathetic neurons depends on cytokines signaling through LIFRbeta containing receptors. AB - Sympathetic ganglia are composed of noradrenergic and cholinergic neurons. Cholinergic sympathetic neurons are characterized by the expression of choline acetyl transferase (ChAT), vesicular acetylcholine transporter (VAChT) and the vasoactive intestinal peptide (VIP). To investigate the role of cytokine growth factor family members in the development of cholinergic sympathetic neurons, we interfered in vivo with the function of the subclass of cytokine receptors that contains LIFRbeta as essential receptor subunit. Expression of LIFRbeta antisense RNA interfered with LIFRbeta expression and strongly reduced the developmental induction of VIP expression. By contrast, ganglion size and the number of ChAT positive cells were not reduced. These results demonstrate a physiological role of cytokines acting through LIFRbeta-containing receptors in the control of VIP expression in sympathetic neurons. PMID- 11880349 TI - Cardiovascular ephrinB2 function is essential for embryonic angiogenesis. AB - EphrinB2, a transmembrane ligand of EphB receptor tyrosine kinases, is specifically expressed in arteries. In ephrinB2 mutant embryos, there is a complete arrest of angiogenesis. However, ephrinB2 expression is not restricted to vascular endothelial cells, and it has been proposed that its essential function may be exerted in adjacent mesenchymal cells. We have generated mice in which ephrinB2 is specifically deleted in the endothelium and endocardium of the developing vasculature and heart. We find that such a vascular-specific deletion of ephrinB2 results in angiogenic remodeling defects identical to those seen in the conventional ephrinB2 mutants. These data indicate that ephrinB2 is required specifically in endothelial and endocardial cells for angiogenesis, and that ephrinB2 expression in perivascular mesenchyme is not sufficient to compensate for the loss of ephrinB2 in these vascular cells. PMID- 11880350 TI - Expression pattern of Brachyury in the mollusc Patella vulgata suggests a conserved role in the establishment of the AP axis in Bilateria. AB - We report the characterisation of a Brachyury ortholog (PvuBra) in the marine gastropod Patella vulgata. In this mollusc, the embryo displays an equal cleavage pattern until the 32-cell stage. There, an inductive event takes place that sets up the bilateral symmetry, by specifying one of the four initially equipotent vegetal macromeres as the posterior pole of all subsequent morphogenesis. This macromere, usually designated as 3D, will subsequently act as an organiser. We show that 3D expresses PvuBra as soon as its fate is determined. As reported for another mollusc (J. D. Lambert and L. M. Nagy (2001) Development 128, 45-56), we found that 3D determination and activity also involve the activation of the MAP kinase ERK, and we further show that PvuBra expression in 3D requires ERK activity. PvuBra expression then rapidly spreads to neighbouring cells that cleave in a bilateral fashion and whose progeny will constitute the posterior edge of the blastopore during gastrulation, suggesting a role for PvuBra in regulating cell movements and cleavage morphology in Patella. Until the completion of gastrulation, PvuBra expression is maintained at the posterior pole, and along the developing anterior-posterior axis. Comparing this expression pattern with what is known in other Bilateria, we advocate that Brachyury might have a conserved role in the regulation of anterior-posterior patterning among Bilateria, through the maintenance of a posterior growth zone, suggesting that a teloblastic mode of axis formation might be ancestral to the Bilateria. PMID- 11880351 TI - The genetic network of prototypic planarian eye regeneration is Pax6 independent. AB - We report the presence of two Pax6-related genes, Pax6A and Pax6B, which are highly conserved in two planarian species Dugesia japonica and Girardia tigrina (Platyhelminthes, Tricladida). Pax6A is more similar to other Pax6 proteins than Pax6B, which is the most divergent Pax6 described so far. The planarian Pax6 homologs do not show any clear orthology to the Drosophila duplicated Pax6 genes, eyeless and twin of eyeless, which suggests an independent Pax6 duplication in a triclad or platyhelminth ancestor. Pax6A is expressed in the central nervous system of intact planarians, labeling a subset of cells of both cephalic ganglia and nerve cords, and is activated during cephalic regeneration. Pax6B follows a similar pattern, but shows a lower level of expression. Pax6A and Pax6B transcripts are detected in visual cells only at the ultrastructural level, probably because a limited amount of transcripts is present in these cells. Inactivation of both Pax6A and Pax6B by RNA-mediated gene interference (RNAi) inhibits neither eye regeneration nor eye maintenance, suggesting that the genetic network that controls this process is not triggered by Pax6 in planarians. PMID- 11880352 TI - BDNF stimulates migration of cerebellar granule cells. AB - During development of the nervous system, neural progenitors arise in proliferative zones, then exit the cell cycle and migrate away from these zones. Here we show that migration of cerebellar granule cells out of their proliferative zone, the external granule cell layer (EGL), is impaired in Bdnf(-/ ) mice. The reason for impaired migration is that BDNF directly and acutely stimulates granule cell migration. Purified Bdnf(-/-) granule cells show defects in initiation of migration along glial fibers and in Boyden chamber assays. This phenotype can be rescued by exogenous BDNF. Using time-lapse video microscopy we find that BDNF is acutely motogenic as it stimulates migration of individual granule cells immediately after addition. The stimulation of migration reflects both a chemokinetic and chemotactic effect of BDNF. Collectively, these data demonstrate that BDNF is directly motogenic for granule cells and provides a directional cue promoting migration from the EGL to the internal granule cell layer (IGL). PMID- 11880353 TI - Larval optic nerve and adult extra-retinal photoreceptors sequentially associate with clock neurons during Drosophila brain development. AB - The visual system is one of the input pathways for light into the circadian clock of the Drosophila brain. In particular, extra-retinal visual structures have been proposed to play a role in both larval and adult circadian photoreception. We have analyzed the interactions between extra-retinal structures of the visual system and the clock neurons during brain development. We first show that the larval optic nerve, or Bolwig nerve, already contacts clock cells (the lateral neurons) in the embryonic brain. Analysis of visual system-defective genotypes showed that the absence of the afferent Bolwig nerve resulted in a severe reduction of the lateral neurons dendritic arborization, and that the inhibition of nerve activity induced alterations of the dendritic morphology. During wild type development, the loss of a functional Bolwig nerve in the early pupa was also accompanied by remodeling of the arborization of the lateral neurons. Approximately 1.5 days later, visual fibers that came from the Hofbauer-Buchner eyelet, a putative photoreceptive organ for the adult circadian clock, were seen contacting the lateral neurons. Both types of extra-retinal photoreceptors expressed rhodopsins RH5 and RH6, as well as the norpA-encoded phospholipase C. These data strongly suggest a role for RH5 and RH6, as well as NORPA, signaling in both larval and adult extra-retinal circadian photoreception. The Hofbauer Buchner eyelet therefore does not appear to account for the previously described norpA-independent light input to the adult clock. This supports the existence of yet uncharacterized photoreceptive structures in Drosophila. PMID- 11880354 TI - Calmodulin-dependent protein kinase IV mediated antagonism of BMP signaling regulates lineage and survival of hematopoietic progenitors. AB - In the current study, we show that bone morphogenetic proteins (BMPs) play a role in hematopoiesis that is independent of their function in specifying ventral mesodermal fate. When BMP activity is upregulated or inhibited in Xenopus embryos hematopoietic precursors are specified properly but few mature erythrocytes are generated. Distinct cellular defects underlie this loss of erythrocytes: inhibition of BMP activity induces erythroid precursors to undergo apoptotic cell death, whereas constitutive activation of BMPs causes an increase in commitment of hematopoietic progenitors to myeloid differentiation and a concomitant decrease in erythrocytes that is not due to enhanced apoptosis. These blood defects are observed even when BMP activity is misregulated solely in non hematopoietic (ectodermal) cells, demonstrating that BMPs generate extrinsic signals that regulate hematopoiesis independent of mesodermal patterning. Further analysis revealed that endogenous calmodulin-dependent protein kinase IV (CaM KIV) is required to negatively modulate hematopoietic functions of BMPs downstream of receptor activation. Our data are consistent with a model in which CaM KIV inhibits BMP signals by activating a substrate, possibly cAMP-response element-binding protein (CREB), that recruits limiting amounts of CREB binding protein (CBP) away from transcriptional complexes functioning downstream of BMPs. PMID- 11880355 TI - reaper is required for neuroblast apoptosis during Drosophila development. AB - Developmentally regulated apoptosis in Drosophila requires the activity of the reaper (rpr), grim and head involution defective (hid) genes. The expression of these genes is differentially regulated, suggesting that there are distinct requirements for their proapoptotic activity in response to diverse developmental and environmental inputs. To examine this hypothesis, a mutation that removes the rpr gene was generated. In flies that lack rpr function, most developmental apoptosis was unaffected. However, the central nervous systems of rpr null flies were very enlarged. This was due to the inappropriate survival of both larval neurons and neuroblasts. Importantly, neuroblasts rescued from apoptosis remained functional, continuing to proliferate and generating many extra neurons. Males mutant for rpr exhibited behavioral defects resulting in sterility. Although both the ecdysone hormone receptor complex and p53 directly regulate rpr transcription, rpr was found to play a limited role in inducing apoptosis in response to either of these signals. PMID- 11880356 TI - Requirement for downregulation of kreisler during late patterning of the hindbrain. AB - Pattern formation in the hindbrain is governed by a segmentation process that provides the basis for the organisation of cranial motor nerves. A cascade of transcriptional activators, including the bZIP transcription factor encoded by the kreisler gene controls this segmentation process. In kreisler mutants, r5 fails to form and this correlates with abnormalities in the neuroanatomical organisation of the hindbrain. Studies of Hox gene regulation suggest that kreisler may regulate the identity as well as the formation of r5, but such a role cannot be detected in kreisler mutants since r5 is absent. To gain further insights into the function of kreisler we have generated transgenic mice in which kreisler is ectopically expressed in r3 and for an extended period in r5. In these transgenic mice, the Fgf3, Krox20, Hoxa3 and Hoxb3 genes have ectopic or prolonged expression domains in r3, indicating that it acquires molecular characteristics of r5. Prolonged kreisler expression subsequently causes morphological alterations of r3/r5 that are due to an inhibition of neuronal differentiation and migration from the ventricular zone to form the mantle layer. We find that these alterations in r5 correlate with an arrest of facial branchiomotor neurone migration from r4 into the caudal hindbrain, which is possibly due to the deficiency in the mantle layer through which they normally migrate. We propose that the requirement for the downregulation of segmental kreisler expression prior to neuronal differentiation reflects the stage-specific roles of this gene and its targets. PMID- 11880357 TI - Involvement of a matrix metalloproteinase in MIS-induced cell death during urogenital development. AB - Programmed cell death of the Mullerian duct eliminates the primitive female reproductive tract during normal male sexual differentiation. Mullerian inhibiting substance (MIS or AMH) triggers regression by propagating a BMP-like signaling pathway in the Mullerian mesenchyme that culminates in apoptosis of the Mullerian duct epithelium. Presently, the paracrine signal(s) used in this developmental event are undefined. We have identified a member of the matrix metalloproteinase gene family, Mmp2, as one of the first candidate target genes downstream of the MIS cascade to function as a paracrine death factor in Mullerian duct regression. Consistent with a role in regression, Mmp2 expression was significantly elevated in male but not female Mullerian duct mesenchyme. Furthermore, this sexually dimorphic expression of Mmp2 was extinguished in mice lacking the MIS ligand, suggesting strongly that Mmp2 expression is regulated by MIS signaling. Using rat organ genital ridge organ cultures, we found that inhibition of MMP2 activity prevented MIS-induced regression, whereas activation of MMP2 promoted ligand-independent Mullerian duct regression. Finally, MMP2 antisense experiments resulted in partial blockage of Mullerian duct regression. Based on our findings, we propose that similar to other developmental programs where selective elimination or remodeling of tissues occurs, localized induction of extracellular proteinases is critical for normal male urogenital development. PMID- 11880358 TI - TLP-1 is an asymmetric cell fate determinant that responds to Wnt signals and controls male tail tip morphogenesis in C. elegans. AB - We have isolated mutations defining a new gene, tlp-1, that affect asymmetric cell fates and morphogenesis during the development of the C. elegans tail. tlp-1 mutations cause defects in the specification of asymmetric cell fates in the descendants of the T blast cell, whose polarity is controlled by Wnt signaling and cause abnormal male tail development leading to the formation of a posterior protrusion reminiscent of 'leptoderan', or pointy tailed, nematode species. In wild-type C. elegans males, which have a 'peloderan' or rounded tail, retraction of the tail tip hypodermis involves a temporally ordered set of cell fusions and changes in cell shape that appear to be heterochronically delayed in tlp-1 males, suggesting that subtle changes in these events can bring about evolutionary changes in morphology. tlp-1 encodes a C2H2 zinc-finger protein that is a member of the Sp family of transcription factors. A TLP-1::GFP fusion protein is expressed in the nuclei of many cells during early embryogenesis and then becomes restricted primarily to posterior cells. At hatching, it is expressed in several head neurons, the posterior intestine cells, tail hypodermal cells, the T cells and specific T-cell descendents in a pattern that suggests TLP-1 may be asymmetrically expressed during the divisions of the T cell lineage. Furthermore, the asymmetry of TLP-1 expression and function appears to be controlled by Wnt signals that control T cell polarity. These results suggest that tlp-1 encodes a transcription factor required for cellular asymmetry that functions downstream of Wnt signals that control cell polarity, as well as in cell fusion and patterning in the C. elegans tail. PMID- 11880359 TI - The plakin Short Stop and the RhoA GTPase are required for E-cadherin-dependent apical surface remodeling during tracheal tube fusion. AB - Cells in vascular and other tubular networks require apical polarity in order to contact each other properly and to form lumen. As tracheal branches join together in Drosophila melanogaster embryos, specialized cells at the junction form a new E-cadherin-based contact and assemble an associated track of F-actin and the plakin Short Stop (shot). In these fusion cells, the apical surface determinant Discs Lost (Dlt) is subsequently deposited and new lumen forms along the track. In shot mutant embryos, the fusion cells fail to remodel the initial E-cadherin contact, to make an associated F-actin structure and to form lumenal connections between tracheal branches. Shot binding to F-actin and microtubules is required to rescue these defects. This finding has led us to investigate whether other regulators of the F-actin cytoskeleton similarly affect apical cell surface remodeling and lumen formation. Expression of constitutively active RhoA in all tracheal cells mimics the shot phenotype and affects Shot localization in fusion cells. The dominant negative RhoA phenotype suggests that RhoA controls apical surface formation throughout the trachea. We therefore propose that in fusion cells, Shot may function downstream of RhoA to form E-cadherin-associated cytoskeletal structures that are necessary for apical determinant localization. PMID- 11880360 TI - Epithelial morphogenesis in hydra requires de novo expression of extracellular matrix components and matrix metalloproteinases. AB - As a member of the phylum Cnidaria, the body wall of hydra is organized as an epithelium bilayer (ectoderm and endoderm) with an intervening extracellular matrix (ECM). Previous studies have established the general molecular structure of hydra ECM and indicate that it is organized as two subepithelial zones that contain basement membrane components such as laminin and a central fibrous zone that contains interstitial matrix components such as a unique type I fibrillar collagen. Because of its simple structure and high regenerative capacity, hydra has been used as a developmental model to study cell-ECM interaction during epithelial morphogenesis. The current study extends previous studies by focusing on the relationship of ECM biogenesis to epithelial morphogenesis in hydra, as monitored during head regeneration or after simple incision of the epithelium. Histological studies indicated that decapitation or incision of the body column resulted in an immediate retraction of the ECM at the wound site followed by a re fusion of the bilayer within 1 hour. After changes in the morphology of epithelial cells at the regenerating pole, initiation of de novo biogenesis of an ECM began within hours while full reformation of the mature matrix required approximately 2 days. These processes were monitored using probes to three matrix or matrix-associated components: basement membrane-associated hydra laminin beta1 chain (HLM-beta1), interstitial matrix-associated hydra fibrillar collagen (Hcol I) and hydra matrix metalloproteinase (HMMP). While upregulation of mRNA for both HLM-beta1 and Hcol-I occurred by 3 hours, expression of the former was restricted to the endoderm and expression of the latter was restricted to the ectoderm. Upregulation of HMMP mRNA was also associated with the endoderm and its expression paralleled that for HLM-beta1. As monitored by immunofluorescence, HLM beta1 protein first appeared in each of the two subepithelial zones (basal lamina) at about 7 hours, while Hcol-I protein was first observed in the central fibrous zone (interstitial matrix) between 15 and 24 hours. The same temporal and spatial expression pattern for these matrix and matrix-associated components was observed during incision of the body column, thus indicating that these processes are a common feature of the epithelium in hydra. The correlation of loss of the ECM, cell shape changes and subsequent de novo biogenesis of matrix and matrix associated components were all functionally coupled by antisense experiments in which translation of HLM-beta1 and HMMP was blocked and head regeneration was reversibly inhibited. In addition, inhibition of translation of HLM-beta1 caused an inhibition in the appearance of Hcol-I into the ECM, thus suggesting that binding of HLM-beta1 to the basal plasma membrane of ectodermal cells signaled the subsequent discharge of Hcol-I from this cell layer into the newly forming matrix. Given the early divergence of hydra, these studies point to the fundamental importance of cell-ECM interactions during epithelial morphogenesis. PMID- 11880361 TI - FGF10 maintains stem cell compartment in developing mouse incisors. AB - Mouse incisors are regenerative tissues that grow continuously throughout life. The renewal of dental epithelium-producing enamel matrix and/or induction of dentin formation by mesenchymal cells is performed by stem cells that reside in cervical loop of the incisor apex. However, little is known about the mechanisms of stem cell compartment formation. Recently, a mouse incisor was used as a model to show that fibroblast growth factor (FGF) 10 regulates mitogenesis and fate decision of adult stem cells. To further illustrate the role of FGF10 in the formation of the stem cell compartment during tooth organogenesis, we have analyzed incisor development in Fgf10-deficient mice and have examined the effects of neutralizing anti-FGF10 antibody on the developing incisors in organ cultures. The incisor germs of FGF10-null mice proceeded to cap stage normally. However, at a later stage, the cervical loop was not formed. We found that the absence of the cervical loop was due to a divergence in Fgf10 and Fgf3 expression patterns at E16. Furthermore, we estimated the growth of dental epithelium from incisor explants of FGF10-null mice by organ culture. The dental epithelium of FGF10-null mice showed limited growth, although the epithelium of wild-type mice appeared to grow normally. In other experiments, a functional disorder of FGF10, caused by a neutralizing anti-FGF10 antibody, induced apoptosis in the cervical loop of developing mouse incisor cultures. However, recombinant human FGF10 protein rescued the cervical loop from apoptosis. Taken together, these results suggest that FGF10 is a survival factor that maintains the stem cell population in developing incisor germs. PMID- 11880362 TI - Implication of protein kinase C in the regulation of DNA mismatch repair protein expression and function. AB - The DNA mismatch repair (MMR) proteins are essential for the maintenance of genomic stability of human cells. Compared with hereditary or even sporadic carcinomas, MMR gene mutations are very uncommon in leukemia. However, genetic instability, attested by either loss of heterozygosity or microsatellite instability, has been extensively documented in chronic or acute malignant myeloid disorders. This observation suggests that in leukemia some internal or external signals may interfere with MMR protein expression and/or function. We investigated the effects of protein kinase C (PKC) stimulation by 12-O tetradecanoylphorbol-13-acetate (TPA) on MMR protein expression and activity in human myeloid leukemia cell lines. First, we show here that unstimulated U937 cells displayed low level of PKC activity as well as MMR protein expression and activity compared with a panel of myeloid cell lines. Second, treatment of U937 cells with TPA significantly increased (3-5-fold) hMSH2 expression and, to a lesser extent, hMSH6 and hPMS2 expression, correlated to a restoration of MMR function. In addition, diacylglycerol, a physiological PKC agonist, induced a significant increase in hMSH2 expression, whereas chelerythrine or calphostin C, two PKC inhibitors, significantly decreased TPA-induced hMSH2 expression. Reciprocally, treatment of HEL and KG1a cells that exhibited a high level of PKC expression, with chelerythrine significantly decreased hMSH2 and hMSH6 expression. Moreover, the alteration of MMR protein expression paralleled the difference in microsatellite instability and cell sensitivity to 6-thioguanine. Our results suggest that PKC could play a role in regulating MMR protein expression and function in some myeloid leukemia cells. PMID- 11880363 TI - RETRACTED: Ubiquitination and degradation of Cdc25C contribute to As(III)-induced G2/M cell cycle arrest. AB - The first author has retracted this paper because of an overlap with Chen, et al, (2002), PNAS 99: 1990-1995, DOI 10.1073/pnas.032428899. PMID- 11880364 TI - Heme oxygenase-1-derived carbon monoxide requires the activation of transcription factor NF-kappa B to protect endothelial cells from tumor necrosis factor-alpha mediated apoptosis. AB - We have shown that carbon monoxide (CO) generated by heme oxygenase-1 (HO-1) protects endothelial cells (EC) from tumor necrosis alpha (TNF-alpha)-mediated apoptosis. This effect relies on the activation of p38 MAPK. We now demonstrate that HO-1/CO requires the activation of the transcription factor NF-kappaB to exert this anti-apoptotic effect. Our data suggest that EC have basal levels of NF-kappaB activity that sustain the expression of NF-kappaB-dependent anti apoptotic genes required to support the anti-apoptotic effect of HO-1/CO. Over expression of the inhibitor of NF-kappaB alpha (IkappaBalpha) suppresses the anti apoptotic action of HO-1/CO. Reconstitution of NF-kappaB activity, by co expression of IkappaBalpha with different members of the NF-kappaB family, i.e. p65/RelA or p65/RelA plus c-Rel, restores the anti-apoptotic effect of HO-1/CO. Expression of the NF-kappaB family members p65/RelA or p65/RelA with p50 or c-Rel up-regulates the expression of the anti-apoptotic genes A1, A20, c-IAP2, and manganese superoxide dismutase (MnSOD). Inhibition of NF-kappaB activity by over expression of IkappaBalpha suppresses the expression of some of these anti apoptotic genes, i.e. c-IAP2. Under inhibition of NF-kappaB, co-expression of some of these anti-apoptotic genes, i.e. c-IAP2 and A1, restores the anti apoptotic action of HO-1/CO, whereas expression of A20 or MnSOD cannot. The ability of c-IAP2 and/or A1 to restore the anti-apoptotic action of HO-1/CO is abolished when p38 MAPK activation is blocked by over-expression of a p38 MAPK dominant negative mutant. In conclusion, we demonstrate that HO-1/CO cooperates with NF-kappaB-dependent anti-apoptotic genes, i.e. c-IAP2 and A1, to protect EC from TNF-alpha-mediated apoptosis. This effect is dependent on the ability of HO 1/CO to activate the p38 MAPK signal transduction pathway. PMID- 11880366 TI - Delineation of the key amino acids involved in neutrophil inhibitory factor binding to the I-domain supports a mosaic model for the capacity of integrin alphaMbeta 2 to recognize multiple ligands. AB - To gain insight into the mechanism by which the alpha(M)I-domain of integrin alpha(M)beta(2) interacts with multiple and unrelated ligands, the identity of the neutrophil inhibitory factor (NIF) recognition site was sought. A systematic strategy in which individual amino acid residues within three previously implicated segments were changed to those in the alpha(L)I-domain, which is structurally very similar but does not bind NIF, was implemented. The capacity of the resulting mutants, expressed as glutathione S-transferase fusion proteins, to recognize NIF was assessed. These analyses ultimately identified Asp(149), Arg(151), Gly(207), Tyr(252), and Glu(258) as critical for NIF binding. Cation binding, a function of the metal ion-dependent adhesion site (MIDAS) motif, was assessed by terbium luminescence to evaluate conformational perturbations induced by the mutations. All five mutants bound terbium with unaltered affinities. When the five residues were inserted into the alpha(L)I-domain, the chimera bound NIF with high affinity. Another ligand of alpha(M)beta(2), C3bi, which is known to use the same segments of the alpha(M)I-domain in engaging the receptor, failed to bind to the chimeric alpha(L)I-domain. Thus, the alpha(M)I-domain appears to present a mosaic of exposed amino acids within surface loops on its MIDAS face, and different ligands interact with different residues to attain high affinity binding. PMID- 11880365 TI - The novel triterpenoid 2-cyano-3,12-dioxooleana-1,9-dien-28-oic acid (CDDO) potently enhances apoptosis induced by tumor necrosis factor in human leukemia cells. AB - Tumor necrosis factor (TNF) is a potent activator of the nuclear factor-kappaB (NF-kappaB) pathway that leads to up-regulation of anti-apoptotic proteins. Hence, TNF induces apoptosis in the presence of inhibitors of protein or RNA synthesis. We report that a novel triterpenoid, 2-cyano-3,12-dioxooleana-1,9, dien-28-oic acid (CDDO) inhibits NF-kappaB-mediated gene expression at a step after translocation of activated NF-kappaB to the nucleus. This effect appears specific for the NF-kappaB pathway as CDDO does not inhibit gene expression induced by the phorbol ester 12-0-tetradecanoylphorbol-13-acetate (TPA). CDDO in combination with TNF caused a dramatic increase in apoptosis in ML-1 leukemia cells that was associated with activation of caspase-8, cleavage of Bid, translocation of Bax, cytochrome c release, and caspase-3 activation. Experiments with caspase inhibitors demonstrated that caspase-8 was an initiator of this pathway. TNF also induced a transient activation of c-Jun N-terminal kinase (JNK), which upon addition of CDDO was converted to a sustained activation. The activation of JNK was also dependent on caspase-8. Sustained activation of JNK is frequently pro-apoptotic, yet inhibition of JNK did not prevent Bax translocation or cytochrome c release, demonstrating its lack of involvement in CDDO/TNF induced apoptosis. Apoptosis was acutely induced by CDDO/TNF in every leukemia cell line tested including those that overexpress Bcl-x(L), suggesting that the mitochondrial pathway is not required for apoptosis by this combination. These results suggest that the apoptotic potency of the CDDO/TNF combination occurs through selective inhibition of NF-kappaB-dependent anti-apoptotic proteins, bypassing potential mitochondrial resistance mechanisms, and thus may provide a basis for the development of novel approaches to the treatment of leukemia. PMID- 11880367 TI - F1-ATPase changes its conformations upon phosphate release. AB - Motor proteins, myosin, and kinesin have gamma-phosphate sensors in the switch II loop that play key roles in conformational changes that support motility. Here we report that a rotary motor, F1-ATPase, also changes its conformations upon phosphate release. The tryptophan mutation was introduced into Arg-333 in the beta subunit of F1-ATPase from thermophilic Bacillus PS3 as a probe of conformational changes. This residue interacts with the switch II loop (residues 308-315) of the beta subunit in a nucleotide-bound conformation. The addition of ATP to the mutant F1 subcomplex alpha3beta(R333W)3gamma caused transient increase and subsequent decay of the Trp fluorescence. The increase was caused by conformational changes on ATP binding. The rate of decay agreed well with that of phosphate release monitored by phosphate-binding protein assays. This is the first evidence that the beta subunit changes its conformation upon phosphate release, which may share a common mechanism of exerting motility with other motor proteins. PMID- 11880368 TI - Loss of ATP-dependent transport activity in pseudoxanthoma elasticum-associated mutants of human ABCC6 (MRP6). AB - Mutations in the ABCC6 (MRP6) gene cause pseudoxanthoma elasticum (PXE), a rare heritable disorder resulting in the calcification of elastic fibers. In the present study a cDNA encoding a full-length normal variant of ABCC6 was amplified from a human kidney cDNA library, and the protein was expressed in Sf9 insect cells. In isolated membranes ATP binding as well as ATP-dependent active transport by ABCC6 was demonstrated. We found that glutathione conjugates, including leukotriene C(4) and N-ethylmaleimide S-glutathione (NEM-GS), were actively transported by human ABCC6. Organic anions (probenecid, benzbromarone, indomethacin), known to interfere with glutathione conjugate transport of human ABCC1 and ABCC2, inhibited the ABCC6-mediated NEM-GS transport in a specific manner, indicating that ABCC6 has a unique substrate specificity. We have also expressed three missense mutant forms of ABCC6, which have recently been shown to cause PXE. MgATP binding was normal in these proteins; ATP-dependent NEM-GS or leukotriene C(4) transport, however, was abolished. Our data indicate that human ABCC6 is a primary active transporter for organic anions. In the three ABCC6 mutant forms examined, the loss of transport activity suggests that these mutations result in a PXE phenotype through a direct influence on the transport activity of this ABC transporter. PMID- 11880369 TI - RasGRP4 is a novel Ras activator isolated from acute myeloid leukemia. AB - Although a number of genetic defects are commonly associated with acute myeloid leukemia (AML), a large percentage of AML cases are cytogenetically normal. This suggests a functional screen for transforming genes is required to identify genetic mutations that are missed by cytogenetic analyses. We utilized a retrovirus-based cDNA expression system to identify transforming genes expressed in cytogenetically normal AML patients. We identified a new member of the Ras guanyl nucleotide-releasing protein (RasGRP) family of Ras guanine nucleotide exchange factors, designating it RasGRP4. Subsequently, cDNA sequences encoding rodent and human RasGRP4 proteins were deposited in GenBank. RasGRP4 contains the same protein domain structure as other members of the RasGRP family, including a Ras exchange motif, a CDC25 homology domain, a C1/diacyglycerol-binding domain, and putative calcium-binding EF hands. We show that expression of RasGRP4 induces anchorage-independent growth of Rat1 fibroblasts. RasGRP4 is a Ras-specific activator and, interestingly, is highly expressed in peripheral blood leukocytes and myeloid cell lines. Unlike other RasGRP proteins, RasGRP4 is not expressed in the brain or in lymphoid cells. We demonstrated that 32D myeloid cells expressing RasGRP4 have elevated levels of activated Ras compared with control cells, and phorbol 12-myristate 13-acetate (PMA) treatment greatly enhanced Ras activation. PMA induced membrane localization of RasGRP4 and 32D cells expressing RasGRP4 were capable of cytokine-independent proliferation in the presence of PMA. We conclude that RasGRP4 is a member of the RasGRP family of Ras guanine nucleotide exchange factors that may play a role in myeloid cell signaling growth regulation pathways that are responsive to diacylglycerol levels. PMID- 11880370 TI - A novel, enzymatically active conformation of the Escherichia coli NADH:ubiquinone oxidoreductase (complex I). AB - Electron microscopy has demonstrated the unusual L-shaped structure of the respiratory complex I consisting of two arms, which are arranged perpendicular to each other. We found that the Escherichia coli complex I has an additional stable conformation, with the two arms arranged side by side, resulting in a horseshoe shaped structure. The structure of both conformations was determined by means of electron microscopy of gold thioglucose-stained single particles. They were distinguished from each other by titration of the complex with polyethylene glycol and by means of analytical ultracentrifugation. The transition between the two conformations is induced by the ionic strength of the buffer and is reversible. Only the horseshoe-shaped complex I exhibits enzyme activity in detergent solution, which is abolished by the addition of salt. Therefore, it is proposed that this structure is the native conformation of the complex in the membrane. PMID- 11880371 TI - Plasmodium falciparum possesses a cell cycle-regulated short type replication protein A large subunit encoded by an unusual transcript. AB - DNA replication in Plasmodium parasites takes place at multiple distinct points during their complex life cycle in the mosquito and vertebrate hosts. Although several parasite proteins involved in DNA replication have been described, the various mechanisms engaged in DNA metabolism of this major pathogen remain largely unexplored. As a step toward understanding this complex network, we describe the identification of Plasmodium falciparum replication protein A large subunit (pfRPA1) through affinity purification and mass spectral analysis of a purified 55-kDa factor. Gel retardation experiments revealed that pfRPA is the major single-stranded DNA binding activity in parasite protein extracts. The activity was expressed in a cell cycle-dependent manner with peak activities in late trophozoites and schizonts, thus correlating with the beginning of chromosomal DNA replication. Accordingly, the pfrpa1 message was detected in parasites 20-24 h post-invasion which is in agreement with the expression of other P. falciparum DNA replication genes. Our results show that pfRPA1 is encoded by an unusual 6.5-kb transcript containing a single open reading frame of which only the C-terminal 42% of the deduced protein sequence shows homologies to other reported RPA1s. Like the orthologues of other protozoan parasites, pfRPA1 lacks the N-terminal protein interaction domain and is thus remarkably smaller than the RPA1s of higher eukaryotes. PMID- 11880372 TI - Ionizing radiation triggers chromatin-bound kin17 complex formation in human cells. AB - The human DNA-binding (HSA)kin17 protein cross-reacts with antibodies raised against the stress-activated Escherichia coli RecA protein. We show here that (HSA)kin17 protein is directly associated with chromosomal DNA as judged by cross linking experiments on living cells. We detected increased amounts of DNA-bound (HSA)kin17 protein 24 h after gamma irradiation, with 2.6-fold more (HSA)kin17 molecules after 6 Gy of irradiation (46,000-117,000 molecules). At this time we observed that highly proliferating RKO cells displayed the concentration and co localization of (HSA)kin17 and replication protein A in nucleoplasmic foci. Our results suggest that 24 h post-irradiation (HSA)kin17 protein may localize at the sites of unrepaired DNA damages. RKO clones expressing an (HSA)KIN17 antisense transcript (RASK.5 and RASK.13 cells) revealed that reduced (HSA)kin17 protein levels are correlated with a decrease in clonogenic cell growth and cell proliferation, as well as an accumulation of cells in early and mid-S phase. Taken together our observations support the idea that (HSA)kin17 protein is a DNA maintenance protein involved in the cellular response to the presence of DNA damage and suggest that it helps to overcome the perturbation of DNA replication produced by unrepaired lesions. PMID- 11880373 TI - Novel mode of action of angiotensin I converting enzyme inhibitors: direct activation of bradykinin B1 receptor. AB - Angiotensin I converting enzyme (kininase II; ACE) inhibitors are important therapeutic agents widely used for treatment in cardiovascular and renal diseases. They inhibit angiotensin II release and bradykinin inactivation; these actions do not explain completely the clinical benefits. We found that enalaprilat and other ACE inhibitors in nanomolar concentrations activate human bradykinin B(1) receptors directly in the absence of ACE and the B(1) agonist des Arg(10)-Lys(1)-bradykinin. These inhibitors activate at the Zn(2+)-binding consensus sequence HEXXH (195-199) in B(1), which is present also in ACE but not in the B(2) receptor. Activation elevates [Ca(2+)](i) and releases NO from endothelial or transfected cells expressing the B(1) receptor but is blocked by Ca-EDTA, a B(1) receptor antagonist, the synthetic undecapeptide sequence (192 202) of B(1), and the mutagenesis of His(195) to Ala(195). Except for the B(1) antagonist, these agents and manipulations did not block activation by a peptide ligand. Thus, Zn(2+) is essential for B(1) receptor activation by ACE inhibitors at the zinc-binding consensus sequence. Ischemia or cytokines induce abundant B(1) receptor expression. B(1) receptor activation by ACE inhibitors, a novel mode of action reported here first, can contribute to their therapeutic effects by releasing NO in the heart and to some side effects. PMID- 11880374 TI - Uncoupled ATP hydrolysis and thermogenic activity of the sarcoplasmic reticulum Ca2+-ATPase: coupling effects of dimethyl sulfoxide and low temperature. AB - The sarcoplasmic reticulum Ca(2+)-ATPase transports Ca(2+) using the energy derived from ATP hydrolysis. During catalysis, part of the energy is used to translocate Ca(2+) across the membrane, and part is dissipated as heat. At 35 degrees C the heat released during the hydrolysis of each ATP molecule varies depending on the formation of a Ca(2+) gradient across the membrane. With leaky vesicles (no gradient) the heat released varies between 9 and 12 kcal/mol of ATP cleaved, and with intact vesicles (gradient), the heat released increases to 20 24 kcal/mol of ATP. After Ca(2+) accumulation, 82% of the Ca(2+)-ATPase activity is not coupled to Ca(2+) transport, and the ratio between Ca(2+) transported and ATP cleaved is 0.3. The addition of 20% dimethyl sulfoxide (v/v) to the medium or decreasing the temperature from 35 to 20 degrees C abolishes the difference of heat produced during ATP hydrolysis in the presence and absence of a gradient. This is accompanied by a simultaneous inhibition of the uncoupled ATPase activity and an increase of the Ca(2+)/ATP ratio from 0.3 to 1.3-1.4. It is concluded that the uncoupled Ca(2+)-ATPase is responsible for both the low Ca(2+)/ATP ratio measured during transport and the difference of heat produced during ATP hydrolysis in the presence and absence of a gradient. PMID- 11880375 TI - Isolation, characterization, and antimicrobial properties of bovine oligosaccharide-binding protein. A microbicidal granule protein of eosinophils and neutrophils. AB - Peptidoglycan recognition proteins (PGRPs) constitute a recently characterized family of pattern-recognition molecules that are conserved from insects to humans and are implicated in mammalian innate immunity. Here we report the isolation, characterization, cDNA cloning, and antimicrobial activities of a bovine PGRP ortholog termed bovine oligosaccharide-binding protein (bOBP). Milligram quantities of bOBP were purified from peripheral leukocytes, thus allowing for the characterization of the disulfide array and for determining the in vitro antimicrobial activities of the native protein. Of the tissues analyzed, bOBP mRNA was detected only in bone marrow where the protein is synthesized as a 190 amino acid precursor. The mature 169 amino acid protein is stored in the cytoplasmic granules of neutrophils and eosinophils but is absent from lymphocytes, monocytes, and platelets. bOBP was microbicidal for Gram-positive and Gram-negative bacteria and yeast at low micromolar concentrations. The finding that bOBP was microbicidal for organisms in which peptidoglycan is absent (Cryptococcus neoformans) or buried (Salmonella typhimurium) indicates that previous conclusions about the specificity of peptidoglycan recognition proteins must be reevaluated and suggests that other envelope components may mediate the antimicrobial action of PGRP family members. PMID- 11880376 TI - Mutant Neuroserpin (S49P) that causes familial encephalopathy with neuroserpin inclusion bodies is a poor proteinase inhibitor and readily forms polymers in vitro. AB - Familial encephalopathy with neuroserpin inclusion bodies (FENIB) is an autosomal dominant dementia that is characterized by intraneuronal inclusions of mutant neuroserpin. We report here the expression, purification, and characterization of wild-type neuroserpin and neuroserpin containing the S49P mutation that causes FENIB. Wild-type neuroserpin formed SDS-stable complexes with tPA with an association rate constant and K(i) of 1.2 x 10(4) m(-1) s(-1) and 5.8 nm, respectively. In contrast, S49P neuroserpin formed unstable complexes with an association rate constant and K(i) of 0.3 x 10(4) m(-1) s(-1) and 533.3 nm, respectively. An assessment by circular dichroism showed that S49P neuroserpin had a lower melting temperature than wild-type protein (49.9 and 56.6 degrees C, respectively) and more readily formed loop-sheet polymers under physiological conditions. Neither the wild-type nor S49P neuroserpin accepted the P7-P2 alpha(1)-anti-trypsin or P14-P3 antithrombin-reactive loop peptides that have been shown to block polymer formation in other members of the serpin superfamily. Taken together, these data demonstrate that S49P neuroserpin is a poor proteinase inhibitor and readily forms loop-sheet polymers. These findings provide strong support for the role of neuroserpin polymerization in the formation of the intraneuronal inclusions that are characteristic of FENIB. PMID- 11880377 TI - Human fragile site FRA16B DNA excludes nucleosomes in the presence of distamycin. AB - Human fragile sites are weak staining gaps in chromosomes generated by specific culture conditions. The short CGG repeating DNA derived from folate-sensitive fragile sites has been shown to exclude single nucleosomes. To test whether this nucleosome exclusion model provides a general molecular mechanism for the formation of fragile sites, a different class of fragile site, the 33-base pair AT-rich repeating DNAs derived from the rare distamycin-inducible site, FRA16B, was examined for its ability to assemble single nucleosomes and nucleosome arrays using in vitro nucleosome reconstitution methods. The FRA16B DNA fragments strongly exclude nucleosome assembly only in the presence of distamycin, and increasing the number of 33-bp repeats increases the effect of distamycin in the destabilization of the nucleosome formation, suggesting a common mechanism for the formation of fragile sites. PMID- 11880378 TI - Impaired stratum corneum hydration in mice lacking epidermal water channel aquaporin-3. AB - The water and solute transporting properties of the epidermis have been proposed to be important determinants of skin moisture content and barrier properties. The water/small solute-transporting protein aquaporin-3 (AQP3) was found by immunofluorescence and immunogold electron microscopy to be expressed at the plasma membrane of epidermal keratinocytes in mouse skin. We studied the role of AQP3 in stratum corneum (SC) hydration by comparative measurements in wild-type and AQP3 null mice generated in a hairless SKH1 genetic background. The hairless AQP3 null mice had normal perinatal survival, growth, and serum chemistries but were polyuric because of defective urinary concentrating ability. AQP3 deletion resulted in a > 4-fold reduced osmotic water permeability and > 2-fold reduced glycerol permeability in epidermis. Epidermal, dermal, and SC thickness and morphology were not grossly affected by AQP3 deletion. Surface conductance measurements showed remarkably reduced SC water content in AQP3 null mice in the hairless genetic background (165 +/- 10 versus 269 +/- 12 microsiemens (microS), p < 0.001), as well as in a CD1 genetic background (209 +/- 21 versus 469 +/- 11 microS). Reduced SC hydration was seen from 3 days after birth. SC hydration in hairless wild-type and AQP3 null mice was reduced to comparable levels (90-100 microS) after a 24-h exposure to a dry atmosphere, but the difference was increased when surface evaporation was prevented by occlusion or exposure to a humidified atmosphere (179 +/- 13 versus 441 +/- 34 microS). Conductance measurements after serial tape stripping suggested reduced water content throughout the SC in AQP3 null mice. Water sorption-desorption experiments indicated reduced water holding capacity in the SC of AQP3 null mice. The impaired skin hydration in AQP3 null mice provides the first functional evidence for the involvement of AQP3 in skin physiology. Modulation of AQP3 expression or function may thus alter epidermal moisture content and water loss in skin diseases. PMID- 11880380 TI - Identification of essential regions in the cytoplasmic tail of interleukin-1 receptor accessory protein critical for interleukin-1 signaling. AB - Interleukin (IL)-1 plays an important role in inflammation and regulation of immune responses. The activated IL-1 receptor complex, which consists of the IL-1 receptor type I and the IL-1 receptor accessory protein (IL-1RAcP), generates multiple cellular responses including NF-kappaB activation, IL-2 secretion, and IL-2 promoter activation. Reconstitution experiments in EL4D6/76 cells lacking IL 1RAcP expression and IL-1 responsiveness were used to analyze structure-function relationships of the IL-1RAcP cytoplasmic tail. Mutating a potential tyrosine kinase phosphorylation motif and various conserved amino acid (aa) residues had no effect on IL-1 responsiveness. Truncation analyses revealed that box 3 of the TIR domain was required for NF-kappaB activation, IL-2 production, and c-Jun N terminal kinase (JNK) activation, whereas IL-2 promoter activation was only partially inhibited. Surprisingly, deletion of aa 527-534 resulted in almost complete loss of all IL-1 responsiveness. Replacement of these aa with alanyl residues did not reconstitute NF-kappaB activation, IL-2 production, or JNK activation but partly restored IL-2 promoter activation. Immunoprecipitation data revealed a strong correlation between MyD88 binding with NF-kappaB activation and IL-2 production but not with IL-2 promoter activation. Taken together, our data indicate that box 3 of IL-1RAcP is critical for IL-1-dependent NF-kappaB activation and stabilization of IL-2 mRNA via JNK, whereas aa 527-534 largely contribute to IL-2 promoter activation. PMID- 11880379 TI - Growth-related renal type II Na/Pi cotransporter. AB - Growth is critically dependent on the retention of a variety of nutrients. The kidney contributes to this positive external balance. In the present study, we isolated a cDNA from the human and rat kidney that encodes a growth-related Na(+) dependent inorganic phosphate (P(i)) cotransporter (type IIc). Microinjection of type IIc cRNA into Xenopus oocytes demonstrated sodium-dependent P(i) cotransport activity. Affinity for P(i) was 0.07 mm in 100 mm Na(+). The transport activity was dependent on extracellular pH. In electrophysiological studies, type IIc Na/P(i) cotransport was electroneutral, whereas type IIa was highly electrogenic. In Northern blotting analysis, the type IIc transcript was only expressed in the kidney and highly in weaning animals. In immunohistochemical analysis, the type IIc protein was shown to be localized at the apical membrane of the proximal tubular cells in superficial and midcortical nephrons of weaning rat kidney. Hybrid depletion experiments suggested that type IIc could function as a Na/P(i) cotransporter in weaning animals, but its role is reduced in adults. The finding of the present study suggest that the type IIc is a growth-related renal Na/P(i) cotransporter, which has a high affinity for P(i) and is electroneutral. PMID- 11880381 TI - A role for p53 in maintaining and establishing the quiescence growth arrest in human cells. AB - The p53 tumor suppressor protein induces transient growth arrest or apoptosis in response to genotoxic stress and mediates the irreversible growth arrest of cellular senescence. We present evidence here that p53 also contributes to the reversible, growth factor-dependent arrest of quiescence (G(0)). Microinjection of expression vectors encoding either MDM2 or a pRb-binding mutant of SV40 T antigen, both of which abrogate p53 function, stimulated quiescent normal human fibroblasts to initiate DNA synthesis and were 40-70% as effective as wild-type T antigen. Electrophoretic mobility shift and p53 transactivation assays showed that p53 activity was higher in quiescent and senescent cells compared with proliferating cells. As proliferating cells entered G(0) after growth factor withdrawal, the p53 mRNA level increased, followed by transient accumulation of the protein. Shortly thereafter, the expression (mRNA and protein) of p21, a p53 target gene and effector of cell cycle arrest, increased. Finally, stable expression of the HPV16 E6 oncogene or dominant negative p53 peptide, GSE-22, both of which inhibit p53 function, delayed entry into quiescence following growth factor withdrawal. Our data indicate that p53 is activated during both quiescence and senescence. They further suggest that p53 activity contributes, albeit not exclusively, to the quiescent growth arrest. PMID- 11880382 TI - 3-Hydroxykynurenine transaminase identity with alanine glyoxylate transaminase. A probable detoxification protein in Aedes aegypti. AB - This study describes the functional characterization of a specific mosquito transaminase responsible for catalyzing the transamination of 3-hydroxykynurenine (3-HK) to xanthurenic acid (XA). The enzyme was purified from Aedes aegypti larvae by ammonium sulfate fractionation, heat treatment, and various chromatographic techniques, plus non-denaturing electrophoresis. The purified transaminase has a relative molecular mass of 42,500 by SDS-PAGE. N-terminal and internal sequencing of the purified protein and its tryptic fragments resolved a partial N-terminal sequence of 19 amino acid residues and 3 partial internal peptide sequences with 7, 10, and 7 amino acid residues. Using degenerate primers based on the partial internal sequences for PCR amplification and cDNA library screening, a full-length cDNA clone with a 1,167-bp open reading frame was isolated. Its deduced amino acid sequence consists of 389 amino acid residues with a predicted molecular mass of 43,239 and shares 45-46% sequence identity with mammalian alanine glyoxylate transaminases. Northern analysis shows the active transcription of the enzyme in larvae and developing eggs. Substrate specificity analysis of this mosquito transaminase demonstrates that the enzyme is active with 3-HK, kynurenine, or alanine substrates. The enzyme has greater affinity and catalytic efficiency for 3-HK than for kynurenine and alanine. The biochemical characteristics of the enzyme in conjunction with the profiles of 3 HK transaminase activity and XA accumulation during mosquito development clearly point out its physiological function in the 3-HK to XA pathway. Our data suggest that the mosquito transaminase was evolved in a manner precisely reflecting the physiological requirement of detoxifying 3-HK produced in the tryptophan oxidation pathway in the mosquito. PMID- 11880383 TI - Analysis of tryptophanase operon expression in vitro: accumulation of TnaC peptidyl-tRNA in a release factor 2-depleted S-30 extract prevents Rho factor action, simulating induction. AB - Expression of the tryptophanase (tna) operon in Escherichia coli is regulated by catabolite repression and tryptophan-induced transcription antitermination. The key feature of this antitermination mechanism has been shown to be the retention of uncleaved TnaC-peptidyl-tRNA in the translating ribosome. This ribosome remains stalled at the tna stop codon and blocks the access of Rho factor to the tna transcript, thereby preventing transcription termination. In normal S-30 preparations, synthesis of a TnaC peptide containing arginine instead of tryptophan at position 12 (Arg(12)-TnaC) was shown to be insensitive to added tryptophan, i.e. Arg(12)-TnaC-peptidyl-tRNA was cleaved, and there was normal Rho dependent transcription termination. When the S-30 extract used was depleted of release factor 2, Arg(12)-TnaC-tRNA(Pro) was accumulated in the absence or presence of added tryptophan. Under these conditions the accumulation of Arg(12) TnaC-tRNA(Pro) prevented Rho-dependent transcription termination, mimicking normal induction. Using a minimal in vitro transcription system consisting of a tna template, RNA polymerase, and Rho, it was shown that RNA sequences immediately adjacent to the tnaC stop codon, the presumed boxA and rut sites, contributed most significantly to Rho-dependent termination. The tna boxA-like sequence appeared to serve as a segment of the Rho "entry" site, despite its likeness to the boxA element. PMID- 11880384 TI - Exploring the stereochemistry of CXCR4-peptide recognition and inhibiting HIV-1 entry with D-peptides derived from chemokines. AB - Chemokine receptor CXCR4 plays an important role in the immune system and the cellular entry of human immunodeficiency virus type 1 (HIV-1). To probe the stereospecificity of the CXCR4-ligand interface, d-amino acid peptides derived from natural chemokines, viral macrophage inflammatory protein II (vMIP-II) and stromal cell-derived factor-1alpha (SDF-1alpha), were synthesized and found to compete with (125)I-SDF-1alpha and monoclonal antibody 12G5 binding to CXCR4 with potency and selectivity comparable with or higher than their l-peptide counterparts. This was surprising because of the profoundly different side chain topologies between d- and l-enantiomers, which circular dichroism spectroscopy showed adopt mirror image conformations. Further direct binding experiments using d-peptide labeled with fluorescein (designated as FAM-DV1) demonstrated that d- and l-peptides shared similar or at least overlapping binding site(s) on the CXCR4 receptor. Structure-activity analyses of related peptide analogs of mixed chiralities or containing alanine replacements revealed specific residues at the N-terminal half of the peptides as key binding determinants. Acting as CXCR4 antagonists and with much higher biological stability than l-counterparts, the d peptides showed significant activity in inhibiting the replication of CXCR4 dependent HIV-1 strains. These results show the remarkable stereochemical flexibility of the CXCR4-peptide interface. Further studies to understand the mechanism of this unusual feature of the CXCR4 binding surface might aid the development of novel CXCR4-binding molecules like the d-peptides that have high affinity and stability. PMID- 11880385 TI - Evidence for distinct cation and calcimimetic compound (NPS 568) recognition domains in the transmembrane regions of the human Ca2+ receptor. AB - The Ca(2+) receptor, a member of the family 3 of G protein-coupled receptors (GPCR), responds not only to its primary physiological ligand Ca(2+) but also to other di- and trivalent metals (Mg(2+), Gd(3+)) and the organic polycations spermine and poly-l-Arginine. As has been found for other family 3 GPCRs, the large amino-terminal extracellular domain (ECD) of the Ca(2+) receptor is the primary Ca(2+) binding domain. To examine how the signal is propagated from the ECD to the seven-transmembrane core domain (7TM) we constructed a Ca(2+) receptor mutant (T903-Rhoc) lacking the entire ECD but containing the 7TM. We have found that this structure initiates signaling in human embryonic kidney (HEK) 293 cells stably expressing the construct. One or more cation recognition sites are also located within the 7TM. Not only Ca(2+), but also several other Ca(2+) receptor specific agonists, Mg(2+), Gd(3+), spermine, and poly-l-Arginine, can activate T903-Rhoc truncated receptor-initiated phosphoinositide hydrolysis in HEK 293 cells. The phenylalkylamine compound, NPS 568, identified as a positive allosteric modulator of the Ca(2+) receptor can selectively potentiate the actions of Ca(2+) and other polycationic agonists on the T903-Rhoc receptor. Similarly, organic polycations synergistically activate T903-Rhoc with di- and trivalent metals. Alanine substitution of all the acidic residues in the second extracellular loop of the T903-Rhoc receptor significantly impairs activation by metal ions and organic polycations in the presence of NPS 568 but not the synergistic activation of Ca(2+) with poly-l-Arginine. These data indicate that although the ECD has been thought to be the main determinant for Ca(2+) recognition, the 7TM core of the Ca(2+) receptor contains activating site(s) recognizing Ca(2+) and Gd(3+) as well as the allosteric modulators NPS 568 and organic polycations that may play important roles in the regulation of receptor activation. PMID- 11880386 TI - Identification of Campylobacter heat-stable and heat-labile antigens by combining the Penner and Lior serotyping schemes. PMID- 11880387 TI - Role of the laboratory in diagnosis and management of cytomegalovirus infection in hematopoietic stem cell and solid-organ transplant recipients. PMID- 11880389 TI - Evaluation of dry and wet transported intravaginal swabs in detection of Chlamydia trachomatis and Neisseria gonorrhoeae infections in female soldiers by PCR. AB - Screening women for sexually transmitted diseases (STD) in nonclinic settings is highly desirable because many infections are asymptomatic. This is especially true for military women, for whom logistical, social, and other job-related obstacles present barriers to accessing medical care. We assessed the accuracy of intravaginal swabs transported by mail in a wet versus a dry state for PCR (Amplicor CT/NG test) detection of chlamydia and gonorrhea infections in a cross sectional study of 793 active-duty military women attending an STD clinic. PCR tests of vaginal swabs (wet and dry) were compared to local clinical methods used on cervical swabs. Standard wet vaginal swab PCR testing detected more chlamydia (11.6%) than cervical enzyme immunoassay (9.3%). For detection of chlamydia using wet swabs, the sensitivity and specificity compared with adjudicated true positives were 94.6% (87 of 92) and 99.3% (696 of 701), respectively. Comparing dry swabs to true-positives for chlamydia, the sensitivity was 91.3% (84 of 92) and the specificity was 99.3% (696 of 701). Standard wet vaginal swab PCR detected more gonorrhea (3.3%) than routine cervical culture (2.1%). The sensitivity and specificity of PCR testing of wet swabs compared to true positives (infected patients) were 96.3% (26 of 27) and 98.2% (752 of 766) for gonorrhea, respectively. For gonorrhea, the sensitivity and specificity of dry swabs compared to true-positives (infected patients) were 88.9% (24 of 27) and 98.3% (753 of 766), respectively. PCR testing of wet and dry transported intravaginal swabs to detect chlamydia and gonorrhea infections was an accurate diagnostic method for military women. PMID- 11880388 TI - Species-level identification of orthopoxviruses with an oligonucleotide microchip. AB - A method for species-specific detection of orthopoxviruses pathogenic for humans and animals is described. The method is based on hybridization of a fluorescently labeled amplified DNA specimen with the oligonucleotide DNA probes immobilized on a microchip (MAGIChip). The probes identify species-specific sites within the crmB gene encoding the viral analogue of tumor necrosis factor receptor, one of the most important determinants of pathogenicity in this genus of viruses. The diagnostic procedure takes 6 h and does not require any sophisticated equipment (a portable fluorescence reader can be used). PMID- 11880390 TI - Incidence of and risk factors for symptomatic visceral leishmaniasis among human immunodeficiency virus type 1-infected patients from Spain in the era of highly active antiretroviral therapy. AB - The way in which the extensive use of highly active antiretroviral therapy (HAART) has influenced the incidence of visceral leishmaniasis (VL) among human immunodeficiency type 1 (HIV-1)-infected patients is not yet understood. The present study assessed whether the incidence of symptomatic VL in HIV-infected patients has decreased since the introduction of HAART. Likewise, the role of other potential risk factors for VL was also analyzed. Therefore, 479 HIV-1 infected patients receiving antiretroviral treatment, according to the available drugs at each moment, were prospectively followed from April 1989 to June 2000 in two university hospitals in southern Spain. A bone marrow aspiration was performed when patients showed symptoms suggestive of kala-azar. A diagnosis of VL was made when Leishmania amastigotes were seen in Giemsa-stained samples or promastigotes were cultured in specific media. The median follow-up time was 1,380 [8 to 4,536] days. Twenty-one patients were diagnosed with symptomatic VL. The density of incidence of VL has decreased 64.8% as of January 1997, when HAART began to be used extensively in our area. The use of HAART was the main independent factor associated with VL; this therapy was a protective factor (adjusted hazard ratio [HR], 0.05; 95% confidence interval [CI], 0.02 to 0.15). CDC clinical category C at entry in the cohort (HR, 4.08; 95% CI, 1.46 to 11.35) and CD4(+) cell counts below 300 cells/mm(3) during the follow-up (HR, 3.96; 95% CI, 1.56 to 10.01) were also independently associated with kala-azar. A VL diagnosis prior to follow-up and low compliance with antiretroviral therapy were not independently associated with symptomatic VL, although statistical significance was almost reached (P = 0.1 and P = 0.08, respectively). In summary, the use of HAART has led to a fall in the incidence of symptomatic VL in HIV infected patients. The main risk factor associated with kala-azar emergence in patients infected with HIV is deep immunosuppression. PMID- 11880391 TI - Performance characteristics of the COBAS Amplicor Hepatitis C Virus (HCV) Monitor, Version 2.0, International Unit assay and the National Genetics Institute HCV Superquant assay. AB - The COBAS Amplicor Hepatitis C Virus (HCV) Monitor assay, version 2.0, which reports in international units per milliliter, was compared to the assay reported in copies per milliliter by analyzing dilution series and clinical plasma samples by both methods. In addition, the Amplicor international unit assay was compared to the National Genetics Institute HCV Superquant assay. The dilution series ranged from <100 to 5,000,000 HCV RNA copies/ml and consisted of 32 points, assayed in triplicate in each assay. Thirty clinical samples ranging from 1,000 to 1,000,000 HCV RNA copies/ml were assayed in duplicate. Deming regression analysis comparing the Amplicor HCV RNA international units-per-milliliter and copies-per-milliliter assays was calculated as follows: (Amplicor international units per milliliter) = 1.030(Amplicor copies per milliliter) - 0.392; R(2) = 0.981; n = 28; S(y/x) (standard error of the estimate) = 0.129. The linearity of the Amplicor international units-per-milliliter assay was as follows: observed = 0.886(expected) + 0.437; R(2) = 0.983; n = 30. The linearity of the Superquant assay was as follows: observed= 0.918 (expected) + 0.436; R(2) = 0.986; n = 32. Deming regression analysis comparing the Amplicor and Superquant assays was calculated as follows: Superquant = 1.066(Amplicor) - 0.0197; R(2) = 0.908; S(y/x) = 0.308; n = 28. The Amplicor and Superquant assays were linear through the range of 600 to 600,000 IU of HCV RNA/ml and approximately 300 to 5,000,000 HCV RNA copies/ml, respectively. The narrow range of the Amplicor assay means that some samples will require dilution and retesting for accurate quantification above 600,000 IU of HCV RNA/ml. The Amplicor and Superquant assays agreed well within the range of 600 to 600,000 IU of HCV RNA/ml (approximately 1,000 to approximately 1,000,000 HCV RNA copies/ml). Overall, the Amplicor and Superquant assays agree well, and results obtained in one assay could be expected to compare well with results from the other when reported in copies per milliliter. PMID- 11880392 TI - Macrolide efflux genes mef(A) and mef(E) are carried by different genetic elements in Streptococcus pneumoniae. AB - Susceptibilities to macrolides were evaluated in 267 Streptococcus pneumoniae isolates, of which 182 were from patients with invasive diseases and 85 were from healthy carriers. Of the 98 resistant isolates, 20 strains showed an M phenotype and carried mef. Strains that carried both mef(A) and mef(E) were found: 17 strains carried mef(A) and 3 carried mef(E). The characteristics of the strains carrying the mef genes and the properties of the mef-containing elements were studied. Strains carrying mef(A) belonged to serotype 14, were susceptible to all the antibiotics tested except erythromycin, and appeared to be clonally related by pulsed-field gel electrophoresis (PFGE). The three mef(E) strains belonged to different serotypes, showed different susceptibility profiles, and did not appear to be related by PFGE. The sequences of a fragment of the mef-containing element, which encompassed mef and the msr(A) homolog, were identical among the three mef(E)-positive strains and among the three mef(A)-positive strains, although there were differences between the sequences for the two variants at 168 positions. In all mef(A)-positive strains, the mef element was inserted in celB, which led to impairment of the competence of the strains. In line with insertion of the mef(E) element at a different site, the competence of the mef(E)-positive strains was maintained. Transfer of erythromycin resistance by conjugation was obtained from two of three mef(A) strains but from none of three mef(E) strains. Due to the important different characteristics of the strains carrying mef(A) or mef(E), we suggest that the distinction between the two genes be maintained. PMID- 11880393 TI - GP5+/6+ PCR followed by reverse line blot analysis enables rapid and high throughput identification of human papillomavirus genotypes. AB - In this study, we developed a simple and fast typing procedure for 37 mucosotropic human papillomavirus (HPV) types using a nonradioactive reverse line blotting (RLB) procedure for general primer (GP5+/6+) PCR products. This system has the advantages not only that in a simple format, up to 42 PCR products can be simultaneously typed per membrane per day, but also that after stripping, the membranes can be easily rehybridized at least 15 times without a loss of signal. RLB appeared highly specific, and its sensitivity was identical to that of conventional typing performed with type-specific oligonucleotide probes in an enzyme immunoassay (EIA). The performance of RLB typing was evaluated with samples of HPV-positive cervical scrapings (n = 196) and biopsies of cervical premalignant lesions (n = 100). The distribution of HPV genotypes detected in these samples was in line with the distribution expected on the basis of literature data. In addition, RLB and EIA typing procedures were compared for the typing of high-risk HPV types in GP5+/6+ PCR products of 210 cervical scrapings from high-risk HPV-positive women who participated in a population-based screening program. The typing procedures had an excellent overall agreement rate of 96.5% (kappa value, 0.77). RLB was successful in detecting multiple HPV infections as well as single infections. In conclusion, the GP5+/6+ PCR-RLB procedure appeared to be a reliable and simple approach that may be of great value for large epidemiological studies, population-based cervical cancer screening programs, and vaccination trials that require high-throughput HPV typing. PMID- 11880394 TI - Performance of the New Bayer VERSANT HCV RNA 3.0 assay for quantitation of hepatitis C virus RNA in plasma and serum: conversion to international units and comparison with the Roche COBAS Amplicor HCV Monitor, Version 2.0, assay. AB - We have evaluated the VERSANT HCV RNA 3.0. Assay (HCV 3.0 bDNA assay) (Bayer Diagnostics, Berkeley, Calif.), which is an improved signal amplification procedure for the HCV 2.0 bDNA assay for the quantitation of hepatitis C virus (HCV) RNA in serum or plasma of HCV-infected individuals. The HCV 3.0 bDNA assay has a linear dynamic range of 2.5 x 10(3) to 4.0 x 10(7) HCV RNA copies per ml (c/ml). The performance of the HCV 3.0 bDNA assay was evaluated using three different test panels. An overall specificity of 96.8% relative to the detection limit of the HCV 3.0 bDNA assay was found. The intra- and interrun reproducibilities for both the dilution panel and the NAP (AcroMetrix, Benicia, Calif.) panel were consistent with coefficients of variation of less than 9%. Quantitation with the HCV 3.0 bDNA assay was linear over the entire range of both panels (ranges of 4.4 x 10(3) to 3.5 x 10(6) c/ml and 5 x 10(3) to 2 x 10(6) IU/ml, respectively), with correlation coefficients of 0.999, slopes close to one, and intercepts close to zero. The regression equation indicated that 1 IU corresponded to about 4.8 copies of HCV RNA. A correlation coefficient of 0.941 was found for HCV RNA values (in international units per milliliter) obtained from the HCV 3.0 bDNA assay and the HCV Monitor version 2.0 assay (HCV Monitor 2.0 assay) (Roche Diagnostic Systems, Branchburg, N.J.). Quantitative results obtained close to the lower limit of the HCV 3.0 bDNA assay might imply that its lower limit should be reconsidered and raised, if necessary. It appeared that quantitation values obtained from the HCV Monitor 2.0 assay of between 5 x 10(2) and 10(5) IU/ml were in general higher than those obtained from the HCV 3.0 bDNA assay, whereas values obtained from the HCV Monitor 2.0 assay were underestimated for samples with HCV RNA levels above 10(5) IU/ml. PMID- 11880395 TI - Food-borne outbreak of gastroenteritis associated with genogroup I calicivirus. AB - An outbreak of gastroenteritis affecting 158 of 219 (72%) guests and employees at a hotel is described. Food served at the hotel restaurant is believed to have been the source of the outbreak and to have been contaminated by sick employees working in the restaurant. A secondary attack rate of 22% was seen involving 43 persons in all. In stool specimens from seven of eight patients, Norwalk-like viruses (NLVs) were detected by electron microscopy. While NLV-specific PCR using primers JV12 and JV13 were negative, all specimens examined with primers NVp69 and NVp110 were positive. The failure of primers JV12 and JV13 was attributed to several mismatches in the JV12 primer. Genotyping and sequence analysis revealed that all samples had identical sequences and clustered with genogroup I, and the most closely related well-characterized genotype is Desert Shield. This is the first described food-borne outbreak associated with genogroup I virus in Sweden. PMID- 11880396 TI - Extended-spectrum beta-lactamase-producing Klebsiella pneumoniae in a neonatal intensive care unit in the high-prevalence area of Athens, Greece. AB - Extended-spectrum beta-lactamase-producing Klebsiella pneumoniae (EPKP) strains are frequently implicated in outbreaks in neonatal intensive care units (NICUs). During the period from 1997 to 1998, 21 infections and 23 colonizations with EPKP were recorded in the NICU of a children's hospital in Athens, Greece. Seventeen of the infected and 12 of the colonized neonates had been referred from other hospitals. The remaining infections and colonizations occurred during the current hospitalization. Pulsed-field gel electrophoresis typing showed that the latter cases were due to an outbreak strain that persisted in the unit, while the repeated introduction of EPKP carriers was mostly due to clonal outbreaks in two maternity hospitals. PMID- 11880397 TI - Streptococcus sinensis sp. nov., a novel species isolated from a patient with infective endocarditis. AB - A bacterium was isolated from the blood culture of a patient with infective endocarditis. The cells were facultative anaerobic, nonsporulating, gram-positive cocci arranged in chains. The bacterium grows on sheep blood agar as alpha hemolytic, gray colonies of 0.5 to 1 mm in diameter after 24 h of incubation at 37 degrees C in ambient air. Growth also occurs in 10 or 40% bile and on bile esculin agar but not in 6% NaCl. No enhancement of growth is observed in 5% CO(2). It is nongroupable with Lancefield groups A, B, C, D, F, or G antisera and is resistant to optochin and bacitracin. The organism is aflagellated and is nonmotile at both 25 and 37 degrees C. It is Voges-Proskauer test positive. It produces leucine arylamidase and beta-glucosidase but not catalase, urease, lysine decarboxylase, or ornithine decarboxylase. It hydrolyzes esculin and arginine. It utilizes glucose, lactose, salicin, sucrose, pullulan, trehalose, cellobiose, hemicellulase, mannose, maltose, and starch. 16S rRNA gene sequencing showed that there were 3.6, 3.7, 4.3, 4.7, and 5.9% differences between the 16S rRNA gene sequence of the bacterium and those of Streptococcus gordonii, Streptococcus intermedius, Streptococcus constellatus, Streptococcus sanguis, and Streptococcus anginosus, respectively. The G+C content of it (mean plus minus standard deviation) was 53.0% plus minus 2.9%. Based on phylogenetic affiliation, it belongs to the mitis or anginosus group of Streptococcus. For these reasons a new species, Streptococcus sinensis sp. nov., is proposed, for which HKU4 is the type strain. Further studies should be performed to ascertain the potential of this bacterium to become an emerging cause of infective endocarditis. PMID- 11880398 TI - Reproducibility of Bordetella pertussis genomic DNA fragments generated by XbaI restriction and resolved by pulsed-field gel electrophoresis. AB - The intra- and interlaboratory variabilities of the molecular size measurements of each DNA fragment contributing to three pulsed-field gel electrophoresis (PFGE) profiles were assessed, as were the reproducibilities of the entire PFGE profiles for three Bordetella pertussis strains. The major source of variability within a laboratory occurred between subcultures rather than within gels or between gels. Each PFGE profile was generated reproducibly and was objectively defined by the molecular sizes of its composite fragments. A strain or profile most suitable for use as an internal reference standard was identified. PMID- 11880399 TI - Diagnosis of respiratory syncytial virus infection: comparison of reverse transcription-PCR to viral culture and serology in adults with respiratory illness. AB - Diagnosis of respiratory syncytial virus (RSV) during acute infection in adults is difficult because of the poor sensitivity of viral culture and antigen detection. A recently developed single-tube nested reverse transcription-PCR (RT PCR) was compared to viral culture and serology by enzyme immunoassay for the diagnosis of RSV in adults with respiratory illness. Nasal swab samples were collected during respiratory illnesses from five groups of subjects: healthy young adults, healthy elderly adults, adults with chronic heart and lung disease, nursing home residents, and adults admitted to the hospital during the winter with cardiopulmonary illnesses. Of 1,112 samples for which all three tests were available, 117 were positive by at least one method and 995 were negative by all methods. One hundred ten were considered true positives because culture and/or serology was positive. Of these, 80 (73%) were PCR positive compared to 43 (39%) that were culture positive. Seven PCR results were considered false positives due to negative culture and serology. The overall RT-PCR sensitivity was 73%, and specificity was 99%. These data indicate that RT-PCR is an excellent method for the diagnosis of acute RSV infection in adults. PMID- 11880400 TI - Association of Bacteroides forsythus and a novel Bacteroides phylotype with periodontitis. AB - Chronic periodontitis is a common infectious disease in the adult population. The etiology is clearly bacterial, and a small number of bacterial species have been consistently associated with periodontitis, including Bacteroides forsythus and Porphyromonas gingivalis. Comparatively little attention has been paid to the identification of health-associated and potentially beneficial bacterial species that may reside in the gingival sulcus. The purpose of the present study was to examine the relationship of the presence of B. forsythus and a newly identified Bacteroides phylotype, oral clone BU063, to periodontal health status. The study was accomplished with a set of samples that were collected from subjects with periodontitis and healthy controls. These samples had previously been analyzed for the presence of P. gingivalis. An oral sampling strategy that included every tooth and a PCR-based detection method were used to maximize detection sensitivity. The presence of B. forsythus in the oral cavity was strongly associated with periodontitis, and its nearest genetic neighbor, oral clone BU063, was associated with oral health (P < 0.0001 for both). Colonization with P. gingivalis was independent of the presence of either Bacteroides species, but the two Bacteroides species were found together less often than would be expected by chance (P < 0.0001). This suggests the presence of a specific exclusionary mechanism between the two Bacteroides species. Comparisons between these two organisms may prove useful for studies that determine how B. forsythus functions in the disease process. In addition, oral clone BU063 deserves further study as a possible preventive or therapeutic intervention for periodontitis. PMID- 11880401 TI - Ca3 fingerprinting of Candida albicans isolates from human immunodeficiency virus positive and healthy individuals reveals a new clade in South Africa. AB - To examine the question of strain specificity in oropharyngeal candidiasis associated with human immunodeficiency virus (HIV) infection, oral samples were collected from 1,196 HIV-positive black South Africans visiting three clinics and 249 Candida albicans isolates were selected for DNA fingerprinting with the complex DNA fingerprinting probe Ca3. A total of 66 C. albicans isolates from healthy black South Africans and 46 from healthy white South Africans were also DNA fingerprinted as controls. Using DENDRON software, a cluster analysis was performed and the identified groups were compared to a test set of isolates from the United States in which three genetic groups (I, II, and III) were previously identified by a variety of genetic fingerprinting methods. All of the characterized South African collections (three from HIV-positive black persons, two from healthy black persons, and one from healthy white persons) included group I, II, and III isolates. In addition, all South African collections included a fourth group (group SA) completely absent in the U.S. collection. The proportion of group SA isolates in HIV-positive and healthy black South Africans was 53% in both cases. The proportion in healthy white South Africans was 33%. In a comparison of HIV-positive patients with and without oropharyngeal symptoms of infection, the same proportions of group I, II, III, and SA isolates were obtained, indicating no shift to a particular group on infection. However, by virtue of its predominance as a commensal and in infections, group SA must be considered the most successful in South Africa. Why group SA isolates represent 53 and 33% of colonizing strains in black and white South Africans and are absent in the U.S. collection represents an interesting epidemiological question. PMID- 11880402 TI - Human immunodeficiency virus type 1 group m protease in cameroon: genetic diversity and protease inhibitor mutational features. AB - To establish a baseline for monitoring resistance to protease inhibitors (PIs) and examining the efficacy of their use among persons in Cameroon infected with human immunodeficiency virus type 1 (HIV-1), we analyzed genetic variability and PI resistance-associated substitutions in PCR-amplified protease (PR) sequences in strains isolated from 110 HIV-1-infected, drug-naive Cameroonians. Of the 110 strains, 85 were classified into six HIV-1 PR subtypes, A (n = 1), B (n = 1), F (n = 4), G (n = 7), H (n = 1), and J (n = 7), and a circulating recombinant form, CRF02-AG (n = 64). PR genes from the remaining 25 (23%) specimens were unclassifiable, whereas 2% (7 of 301) unclassifiable PR sequences were reported for a global collection. Two major PI resistance-associated mutations, 20M and 24I, were detected in strains from only two specimens, whereas secondary mutations were found in strains from all samples except one strain of subtype B and two strains of CRF02-AG. The secondary mutations showed the typical PI resistance-associated pattern for non-subtype B viruses in both classifiable and unclassifiable PR genes, with 36I being the predominant (99%) mutation, followed by 63P (18%), 20R (15%), 77I (13%), and 10I or 10V (11%). Of these mutations, dual and triple PI resistance-associated substitutions were found in 38% of all the Cameroonian strains. Compared with classifiable PR sequences, unclassifiable sequences had significantly more dual and triple substitutions (64% versus 30%; P = 0.004). Phenotypic and clinical evaluations are needed to estimate whether PI resistance during antiretroviral drug treatment occurs more rapidly in individuals infected with HIV-1 strains harboring multiple PI resistance associated substitutions. This information may be important for determination of appropriate drug therapies for HIV-1-infected persons in Cameroon, where more than one-third of HIV-1 strains were found to carry dual and triple minor PI resistance-associated mutations. PMID- 11880403 TI - Burkholderia cepacia complex bacteria from clinical and environmental sources in Italy: genomovar status and distribution of traits related to virulence and transmissibility. AB - Sixty-eight Burkholderia cepacia complex isolates recovered from the sputum of 53 cystic fibrosis patients and 75 isolates collected from the maize rhizosphere were compared to each other to assess their genomovar status as well as some traits related to virulence such as antibiotic susceptibility, proteolytic and hemolytic activities, and transmissibility, in which transmissibility is determined by detection of the esmR and cblA genes. Among the clinical isolates, B. cepacia genomovar III comprised the majority of isolates examined and only a very few isolates were assigned to B. cepacia genomovar I, B. stabilis, and B. pyrrocinia; among the environmental isolates a prevalence of B. cepacia genomovar III and B. ambifaria was observed, whereas few environmental isolates belonging to B. cepacia genomovar I and B. pyrrocinia were found. Antibiotic resistance analysis revealed a certain degree of differentiation between clinical and environmental isolates. Proteolytic activity and onion tissue maceration ability were found to be spread equally among both clinical and environmental isolates, whereas larger percentages of environmental isolates than clinical isolates had hemolytic activity. The esmR gene was found exclusively among isolates belonging to B. cepacia genomovar III, with a marked prevalence in clinical isolates, whereas only one clinical isolate belonging to B. cepacia genomovar III was found to bear the cblA gene. In conclusion, the results of the present study show that the species compositions of the clinical and environmental B. cepacia complex populations examined are quite different and that some of the candidate determinants related to virulence and transmissibility are not confined solely to clinical isolates but are also spread among environmental isolates belonging to different species of the B. cepacia complex. PMID- 11880404 TI - Trends in antifungal susceptibility of Candida spp. isolated from pediatric and adult patients with bloodstream infections: SENTRY Antimicrobial Surveillance Program, 1997 to 2000. AB - From 1 January 1997 through 31 December 2000, 2,047 bloodstream infections (BSIs) due to Candida spp. were reported from hospitals in the United States, Canada, Latin America, and Europe participating in the SENTRY Antifungal Surveillance Program. Among individuals in four age groups (< or =1, 2 to 15, 16 to 64, and > or =65 years) Candida albicans was the most common species, causing 60, 55, 55, and 50% of infections, respectively. C. glabrata caused 17 to 23% of BSIs in those ages 16 to 64 and > or = 65 years, whereas it caused only 3% of BSIs in the individuals in the two younger age groups (P < 0.001). C. parapsilosis (which caused 21 to 24% of BSIs) and C. tropicalis (which caused 7 to 10% of BSIs) were more common than C. glabrata in individuals ages < or =1 year and 2 to 15 years. Isolates of Candida spp. showed a trend of decreasing susceptibility to fluconazole, itraconazole, and amphotericin B with increasing patient age (P < or = 0.01). None of the C. glabrata isolates from individuals < or =1 year old were resistant to fluconazole, whereas they made up 5 to 9% of isolates from individuals ages 16 to 64 and > or =65 years. Isolates of C. tropicalis from patients < or =1 year old were more susceptible to flucytosine (MIC at which 90% of isolates are inhibited [MIC(90)], 0.5 microg/ml; 0% resistant isolates) than those from patients > or =65 years old (MIC(90), 32 microg/ml; 11% resistant isolates). The investigational triazoles posaconazole, ravuconazole, and voriconazole were all highly active against all species of Candida from individuals in all age groups. These data demonstrate differences in the species distributions of pathogens and differences in antifungal resistance among isolates from individuals in the pediatric and adult age groups. Ongoing surveillance will enhance efforts to limit the extent of antifungal resistance in individuals in various age groups. PMID- 11880405 TI - Detection of seg, seh, and sei genes in Staphylococcus aureus isolates and determination of the enterotoxin productivities of S. aureus isolates Harboring seg, seh, or sei genes. AB - To investigate the distribution of staphylococcal enterotoxin (SE) A to I (SEA to SEI) genes (sea to sei) in Staphylococcus aureus, 146 isolates obtained in Japan from humans involved in and samples from food poisoning outbreaks, healthy humans, cows with mastitis, and bovine raw milk were analyzed by multiplex PCR. One hundred thirteen (77.4%) S. aureus isolates were found to be positive for one or more se genes. The se genotype was classified into 14 genotypes. seg and sei coexisted in the same S. aureus strain. The newly developed sandwich enzyme linked immunosorbent assay showed that most seh-harboring S. aureus isolates were able to produce a significant amount of SEH. However, most of the S. aureus isolates harboring seg and about 60% of the isolates harboring sei did not produce a detectable level of SEG or SEI, while reverse transcription-PCR analysis proved that the mRNAs of SEG and SEI were transcribed in S. aureus strains harboring seg and sei genes. These results suggest the importance of quantitative assessment of SEG and SEI production in foods in order to clarify the relationship between these new SEs and food poisoning. PMID- 11880406 TI - Rapid and sensitive detection of physical status of human papillomavirus type 16 DNA by quantitative real-time PCR. AB - A rapid quantitative real-time PCR method was employed to quantify the copy number of E2 and E6 genes for analysis of the physical status of human papillomavirus type 16 (HPV-16) DNA. Significant differences with respect to both copy numbers were found when more than 40% of HPV-16 DNA was integrated with disruption of the E2 gene in an experimental model. The physical status of HPV-16 DNA in 50 clinical samples was exclusively episomal in 21 cases (42%), concomitant in 14 cases (28%), and integrated in 15 cases (30%). The prevalence of integrated and/or concomitant forms of HPV-16 DNA increased with progression of cervical disease. Four of 11 cervical intraepithelial neoplasia involved integrated forms of HPV-16 DNA partially or exclusively. This rapid, sensitive technique is useful in the analysis of the physical status of HPV DNA. PMID- 11880408 TI - Distribution of hepatitis B virus genotypes among American blood donors determined with a PreS2 epitope enzyme-linked immunosorbent assay kit. AB - We genotyped 418 sera from volunteer blood donors from two large, regional blood centers in the United States who were HBsAg positive by an enzyme-linked immunosorbent assay (ELISA). The HBV genotypes were determined by a serological method using a preS2 epitope ELISA kit (Institute of Immunology, Tokyo, Japan) with monoclonal antibodies. Of the 418 samples, the genotypes of 320 could be determined (76.6%). One hundred forty-three (34.2%) were genotyped as A (preS2 subtype su), 31 (7.4%) were genotyped as B (subtype m), 59 (14.1%) were genotyped as C (subtype ks), 83 (19.9%) were genotyped as D or E (subtype ksu), and 4 (1.0%) were genotyped as F (subtype k). This kit cannot differentiate genotypes D and E. For 98 (23.4%) of the 418 samples, the genotype could not be determined; 11 of these 98 samples were positive for at least one of the preS2 genotype specific epitopes (m, k, s, and u), but the combinations of positive epitopes were different from those of samples that could be genotyped; 45 had only the common epitope (b). In the group with a high signal-to-cutoff (S/C) ratio, the HBV genotype could be determined for 199 (84%) of 237 samples; in contrast, in the low-S/C-ratio group, only 10 (20%) of 51 samples could be genotyped (P < 0.001). These findings may indicate the limitation of genotyping samples with low S/C ratios for HBsAg by ELISA or the existence of genotype G or other new HBV genotypes in HBsAg-positive blood donors in the United States. Of the genotyped samples, 201 were assayed for HBeAg; only 9 (4.5%) were positive for HBeAg. The frequency of genotype C in HBeAg-positive donor samples (5 of 9 or 56%) was higher than that in HBeAg-negative donor samples (33 of 192, or 17%) (P = 0.022). PMID- 11880407 TI - Molecular typing of selected Enterococcus faecalis isolates: pilot study using multilocus sequence typing and pulsed-field gel electrophoresis. AB - The present study compared the recently developed multilocus sequence typing (MLST) approach with a well-established molecular typing technique, pulsed-field gel electrophoresis (PFGE), for subspecies differentiation of Enterococcus faecalis isolates. We sequenced intragenic regions of three E. faecalis antigen encoding genes (ace, encoding a collagen and laminin adhesin; efaA, encoding an endocarditis antigen; and salA, encoding a cell wall associated antigen) and one housekeeping gene (pyrC) of 22 E. faecalis isolates chosen largely for their temporal and geographical diversity, but also including some outbreak isolates. MLST analysis of polymorphic regions of these four genes identified 13 distinct sequence types (STs) with different allelic profiles; the composite sequences generated from the four sequenced gene fragments of individual isolates showed 98.3 to 100% identity among the 22 isolates. We also found that the allelic profiles from two sequences, ace and salA, were sufficient to distinguish all 13 STs of this study. The 13 STs corresponded to 12 different PFGE types, with one previously designated PFGE clone (a widespread U.S. clone of beta-lactamase producing isolates) being classified into two highly related STs which differed at 2 of 2,894 bases, both in the same allele. MLST also confirmed the clonal relationships among the isolates of two other PFGE clonal groups, including vancomycin resistant isolates. Thus, this pilot study with representative E. faecalis isolates suggests that, similar to PFGE, the sequence-based typing method may be useful for differentiating isolates of E. faecalis to the subspecies level in addition to identifying outbreak isolates. PMID- 11880409 TI - Diagnostic detection of human immunodeficiency virus type 1 antibodies in urine: a brazilian study. AB - We evaluated, for the first time in Latin America, the performance of a commercial enzyme immunoassay (EIA) (Calypte Biomedical Corporation, Berkeley, Calif.) that detects human immunodeficiency virus type 1 (HIV-1)-specific antibodies in urine in comparison to standard serological assays (two commercial EIAs and a commercial Western blot [WB] assay). Paired serum and urine specimens were collected from two different groups of Brazilian patients: 225 drug users with unknown HIV status who attended drug treatment centers in Rio de Janeiro, Brazil, and 135 subjects with known HIV status. Patients showing positive results in the serum EIAs and/or in the urine EIA were serologically confirmed by WB assay. For 135 individuals with known HIV status, the urine EIA showed 100% sensitivity (74 positive samples) and 95.1% specificity (58 of 61 negative specimens). For 225 drug users, the test showed 100% sensitivity (2 positive samples) and 98.7% specificity (220 of 223 negative samples) compared to WB confirmed serological EIA results. Thus, in a total of 360 samples, the urine EIA correctly identified all 76 HIV-positive samples and 278 of 284 negative samples (100% sensitivity and 97.9% specificity). Detailed analysis of the urine EIA results indicates that an increase of the recommended cutoff value might raise the specificity of the assay without affecting its sensitivity. Our results suggest that the HIV-1 urine EIA is a good screening test suitable for developing countries like Brazil. However, as for all other HIV screening tests on the market, it is not specific enough to be used as a one-step test and therefore requires confirmation. PMID- 11880410 TI - Integrated human papillomavirus type 16 is frequently found in cervical cancer precursors as demonstrated by a novel quantitative real-time PCR technique. AB - In contrast to cervical cancer, integration of human papillomavirus (HPV) DNA into the host genome has been considered a rare event in cancer precursor lesions (cervical intraepithelial neoplasia [CIN]). With our new real-time PCR method, we demonstrated that integrated HPV type 16 (HPV16) is already present in CIN lesions. The physical state of HPV16 and the viral load were simultaneously detected. A unique region of the E2 open reading frame (ORF) that is most often deleted during HPV16 integration is targeted by one set of PCR primers and a probe, and another set targets the E6 ORF. In episomal form, both targets should be equivalent, while in integrated form, the copy numbers of E2 would be less than those of E6. The method was tested with DNAs from 31 cervical lesions (non CIN to CINIII) from 24 women prospectively followed up for 10 years. This report presents viral load and integration results from the largest series of CIN lesions described to date. Only one sample contained exclusively episomal HPV16 DNA, and this lesion regressed spontaneously. Samples from another patient, with only integrated HPV16, rapidly progressed from CINI to CINIII in 2 years. In all other patients, episomal and integrated forms of HPV16 DNA were found to coexist. Rapid progression of the CIN lesions was closely associated with a heavy load of integrated HPV16. Thus, the method described here is a very sensitive tool with which to assess the physical state of HPV, which is useful in predicting disease progression. PMID- 11880412 TI - Initial concentration of Staphylococcus epidermidis in simulated pediatric blood cultures correlates with time to positive results with the automated, continuously monitored BACTEC blood culture system. AB - The relationship of initial concentration of Staphylococcus epidermidis in blood cultures and time to positivity (TTP) in an automated, continuously monitored blood culture system was assessed. Blood and 1 to 1,000 CFU of S. epidermidis per ml in stationary or exponential phase were inoculated in BACTEC Pediatric Plus F bottles and incubated. The TTP was inversely proportional to the initial inoculated concentration. Blood culture bottles with initial bacterial densities of <10 CFU/ml had a TTP of >20 h (upper limit of 95% prediction interval, 20.7 h) and bottles with initial bacterial densities of > or =50 CFU/ml had a TTP of < or =15 h (lower limit of 95% prediction interval, 13.4 h). PMID- 11880411 TI - PCR detection and serological evidence of granulocytic ehrlichial infection in roe deer (Capreolus capreolus) and chamois (Rupicapra rupicapra). AB - The role of wild mammals, such as roe deer (Capreolus capreolus) and chamois (Rupicapra rupicapra), in the epidemiology of granulocytic ehrlichiae in Switzerland was investigated. We tested blood samples for Ehrlichia phagocytophila genogroup 16S rRNA gene sequences by PCR and for immunoglobulin G antibodies against granulocytic ehrlichiae by indirect fluorescent-antibody assay (IFA). Overall means of 60.9% of 133 roe deer serum samples and 28.2% of 39 chamois serum samples were seroreactive by IFA. PCR results were positive for 18.4% of 103 roe deer serum samples as well. None of the 24 chamois blood samples tested were positive by PCR. Partial 16S rRNA gene and groESL heat shock operon sequences of three roe deer samples tested showed strong degrees of homology (> or =99.7 and > or =98.6%, respectively) with the sequences of granulocytic ehrlichiae isolated from humans. These results confirm that chamois, and particularly roe deer, are commonly infected with granulocytic ehrlichiae and provide evidence that these wild mammals are potential reservoirs for granulocytic ehrlichiae in Switzerland. PMID- 11880413 TI - Use of PGMY primers in L1 consensus PCR improves detection of human papillomavirus DNA in genital samples. AB - The novel PGMY L1 consensus primer pair is more sensitive than the MY09 and MY11 primer mix for detection and typing with PCR of human papillomavirus (HPV) DNA in genital specimens. We assessed the diagnostic yield of PGMY primers for the detection and typing of HPV by comparing the results obtained with PGMY09/PGMY11 and MY09/MY11/HMB01 on 299 genital samples. Amplicons generated with PGMY primers were typed with the line blot assay (PGMY-line blot), while HPV amplicons obtained with the degenerate primer pool MY09/MY11/HMB01 were detected with type specific radiolabeled probes in a dot blot assay (standard consensus PCR test). Cervicovaginal lavage samples (N = 272) and cervical scrape samples (N = 27) were tested in parallel with both PCR tests. The PGMY-line blot test detected the presence of HPV DNA more frequently than the standard consensus PCR assay. The concordance for HPV typing between the two assays was 84.3% (214 of 255 samples), for a good kappa value of 0.69. Of the 177 samples containing HPV DNA by at least one method, 40 samples contained at least one HPV type detected only with PGMY line blot, whereas positivity exclusively with the standard consensus PCR test was found for only 7 samples (P < 0.001). HPV types 45 and 52 were especially more frequently detected with PGMY than MY primers. However, most HPV types were better amplified with PGMY primers, including HPV-16. Samples with discordant results between the two PCR assays more frequently contained multiple HPV types. Studies using PGMY instead of MY primers have the potential to report higher detection rates of HPV infection not only for newer HPV types but also for well known genital types. PMID- 11880414 TI - New simple and rapid test for culture confirmation of Mycobacterium tuberculosis complex: a multicenter study. AB - Mycobacterial antigen MPB64 has been identified as a Mycobacterium tuberculoisis complex-specific secretory protein since 1984. Recently, a simple culture confirmation test for M. tuberculosis complex has been developed by using lateral flow immunochromatographic assay (ICA) to detect MPB64 with anti-MPB64 monoclonal antibody. The current multicenter study evaluated the performance of an ICA slide test for MPB64 antigen in the clinical setting. Primary positive cultures from clinical specimens, as well as stock cultures, were tested. Approximately 100 microl of positive liquid culture medium or suspension made from colonies on solid medium was placed into the test well of the plastic slide devise, and the test was read after 15 min. No processing or instrumentation was required. A total of 304 mycobacterial isolates consisting of M. tuberculosis complex (171 isolates) and mycobacteria other than M. tuberculosis (MOTT) complex (133 isolates) belonging to 18 different species were tested. Growth in liquid media (Mycobacteria Growth Indicator Tube [MGIT] and Radiometric 12B), as well as in solid (Lowenstein-Jensen and Middlebrook 7H10 agar) media, was evaluated. Results were compared with those obtained with nucleic acid-based and/or high-pressure liquid chromatography identification. All MOTT were found to be negative on the ICA slide with no cross-reaction. All M. tuberculosis and M. africanum cultures were found to be positive, whereas the results of M. bovis and M. bovis BCG cultures were variable since some of the BCG strains are known to lack MPB64 antigen production. The results did not change with prolonged storage of cultures. This low-tech rapid test with high sensitivity and specificity could provide an alternative to currently available identification methods, particularly for recently introduced nonradiometric liquid culture systems such as MGIT. PMID- 11880415 TI - Enterococcus faecium-related outbreak with molecular evidence of transmission from pigs to humans. AB - Between 24 July and 31 August 1998, thousands of domestic pigs died of hemorrhagic shock in three adjunct counties along the YangZi River in Jiangshu Province, China. From 28 July to 6 September 1998, 40 local farmers (36 males and 4 females, ages 23 to 78 years) were hospitalized with severe illness characterized by high fever, erythematous rash or petechiae, and profound lethargy after contact with sick pigs. Twelve (30%) of these patients died of respiratory failure and shock. Eleven bacterial isolates recovered from 11 blood and cerebrospinal fluid specimens collected from seven patients and two pigs were identified as Enterococcus faecium based on biochemical reactions and 16S rRNA gene sequence analysis. Both pig and human E. faecium isolates displayed indistinguishable antibiotic susceptibility and pulsed-field gel electrophoresis patterns. These data strongly suggest the spread of an outbreak of E. faecium related sepsis from pigs to humans. PMID- 11880416 TI - Susceptibility testing of fluconazole by the NCCLS broth macrodilution method, E test, and disk diffusion for application in the routine laboratory. AB - Antifungal susceptibility testing may be an important aid in the treatment of patients with life-threatening yeast infections. In order to establish the suitability of different susceptibility test methods for fluconazole with yeasts, the Rosco tablet and the E-test were compared with the gold standard NCCLS broth macrodilution method for 106 yeast strains. These included 102 clinical isolates of Candida spp., including Candida glabrata (n = 30), Candida albicans (n = 20), Candida tropicalis (n = 13), Candida parapsilosis (n = 10), Candida krusei (n = 8), plus Cryptococcus neoformans (n = 3), Saccharomyces cerevisiae (n = 2), and 16 strains belonging to other Candida spp. Four American Type Culture Collection strains of Candida were included as quality controls. The NCCLS method was found to be too complex and labor-intensive for routine testing. The E-test is an accurate alternative, but experience in determining MICs and careful attention to procedural details are critically important. The Rosco tablet showed the best agreement with the NCCLS reference method, especially when newly established breakpoints of R < or = 10 mm and S > or = 21 mm were used. PMID- 11880417 TI - Characterization of Shiga toxin-producing Escherichia coli O26 strains and establishment of selective isolation media for these strains. AB - We characterized the carbohydrate-fermenting ability of 31 strains of Shiga toxin producing Escherichia coli (STEC) O26 isolated from diarrhea patients in Aichi Prefecture, Japan, in order to establish selective isolation media for these strains. None of the 31 STEC O26 strains (24 O26:H11, 7 O26:H-) fermented rhamnose, whereas all of the other 108 STEC strains (100 O157, 8 O111) and all of the non-STEC strains except one (i.e., 58 of 59) fermented rhamnose. The great majority of the STEC O26 strains (96.8% [30 of 31]) showed very high resistance to potassium tellurite (MIC > or = 50 microg/ml), whereas the majority of the non STEC strains (72.9% [43 of 59]) showed very high sensitivity (MIC < or = 1.56 microg/ml) to this compound. Accordingly, we developed a rhamnose-MacConkey (RMAC) medium in which the lactose in MacConkey medium was replaced by rhamnose, and cefixime-tellurite-RMAC (CT-RMAC) medium in which potassium tellurite (2.5 mg/liter) and cefixime (0.05 mg/liter) were added to RMAC. All of the STEC O26 strains generated colorless (rhamnose-nonfermented) colonies on both media; the vast majority of selected E. coli strains (95.7% [89 of 93; including 26 STEC O157, 8 STEC O111]), other than STEC O26, generated red colonies on RMAC, and most of the non-STEC strains (84.7% [50 of 59]) did not grow on CT-RMAC. We demonstrate that both the RMAC and the CT-RMAC media can be used for the isolation of STEC O26 and that CT-RMAC has better specificity for the routine isolation for STEC O26 in a laboratory. PMID- 11880419 TI - Prevalence of parvovirus B19 and parvovirus V9 DNA and antibodies in paired bone marrow and serum samples from healthy individuals. AB - Parvovirus B19 (hereafter referred to as B19) exhibits a marked tropism to human bone marrow (BM), and infection may lead to erythema infectiosum, arthropathy, hydrops fetalis, and various hematologic disorders. Recently, a distinct parvovirus isolate termed V9 with an unknown clinical spectrum was discovered. In contrast to the many studies of B19 serology and viremia, valid information on the frequency of B19 or V9 DNA in the BM of healthy individuals is limited. To develop a reference value, paired BM and serum samples from healthy subjects were tested for the presence of B19 and V9 DNA and specific antibodies. Immunoglobulin M (IgM) was not found in any of the serum samples. The prevalence of IgG showed a gradual and steady increase from 37% in children aged 1 to 5 years to 87% in people aged >50 years. When 190 well-characterized subjects were examined, B19 DNA was detected in the BM of 4 individuals (2.1%; 95% confidence interval, 0.58 to 5.3%) while none of the paired serum samples showed evidence of circulating viral DNA. V9 DNA was not found in any of the BM or serum samples. The finding of B19 DNA probably indicated a primary infection in one 7-year-old individual and reinfection or reactivation of persistent infection in the remaining three persons, aged 47 to 58 years. Serving as a benchmark for future studies, these findings are useful when interpreting epidemiologic data, performing BM transplantation, or considering clinical implications of parvovirus infection. PMID- 11880418 TI - Detection of neutralizing antibodies against human papillomaviruses (HPV) by inhibition of gene transfer mediated by HPV pseudovirions. AB - The goal of this study was to develop a human papillomavirus (HPV) neutralization assay using HPV pseudovirions generated in vitro. For this purpose, gene transfer efficiency of HPV virus-like particles (VLPs) was improved by using direct interaction between a reporter plasmid and the VLPs. Electron microscopic observation of the interaction between DNA molecules and VLPs revealed that VLPs always interact with a single DNA molecule and that VLPs bind to the end of linearized DNA molecules. An 100-fold improvement in the gene transfer was obtained by simple interaction between a linearized DNA molecule and VLPs. Moreover, direct interaction methods offer the possibility of transferring plasmids a size higher than that of the papillomavirus genome. The approach that we developed to generate HPV-16 and HPV-31 pseudovirions proved to be suitable for testing neutralizing antibodies in human sera both after immunization and after natural infection. PMID- 11880420 TI - Identification and molecular characterization of a bovine G3 rotavirus which causes age-independent diarrhea in cattle. AB - G3 rotaviruses have been reported rarely in cattle, and none have been characterized. We report the first genomic characterization of a bovine G3 rotavirus, CP-1, which had been biologically characterized in vivo and shown to cause age-independent diarrhea. CP-1 was a G3 rotavirus as its VP7 had 92 to 96% deduced amino acid identity to those of G3 rotaviruses. However, initially, CP-1 was identified as a G10 rotavirus by RT-PCR even though the CP-1 VP7 had only 81 to 85% deduced amino acid identity to those of G10 rotaviruses. Rotavirus CP-1 was of P[5] specificity, a type common in cattle, and had a bovine NSP1 and NSP4. These results added another animal species to those in which G3 rotaviruses have been found, characterized a bovine rotavirus which caused age-independent diarrhea in calves, and raised the possibility that bovine G3 rotaviruses may be misdiagnosed as G10 rotaviruses. PMID- 11880421 TI - Specificities and functions of the recA and pps1 intein genes of Mycobacterium tuberculosis and application for diagnosis of tuberculosis. AB - The worldwide recrudescence of tuberculosis and the widespread appearance of antibiotic resistance have strengthened the need for rapid and specific diagnostic tools. The prevailing microbiological identification of Mycobacterium tuberculosis, the causative agent of tuberculosis, which implies the use of in vitro cultures and acid-fast staining microscopy, is time-consuming. Detection of M. tuberculosis directly in clinical samples through PCR amplification of mycobacterium-specific genes, designed to shorten diagnostic delay, demonstrated reliability and high sensitivity. However, the quality of the diagnosis depends on the specificity of the target sequence for M. tuberculosis complex strains. In the present study, we demonstrated the specificity of recA and pps1 inteins for this complex and thus the feasibility of using intein-coding sequences as a new target for PCR diagnosis. Indeed, the recA and pps1 genes of 36 clinical isolates of M. tuberculosis and 10 field strains of M. bovis were found to be interrupted by an intein sequence at the RecA-a and Pps1-b sites, respectively, while a large number of nontuberculous mycobacterial species failed to demonstrate these insertions. Besides, the MtuPps1, which was cloned and expressed in Escherichia coli, was shown to possess an endonuclease activity. The intein cleaves the 40-bp sequence spanning the intein insertion site Pps1-b in the inteinless pps1 gene. In addition to the PCR amplification of recA and pps1 intein genes as a tool for diagnosis, the specific endonuclease activity could represent a new molecular approach to identify M. tuberculosis. PMID- 11880422 TI - Antigenic diversity and distribution of rabies virus in Mexico. AB - Rabies remains a public health problem in the Americas because of the great diversity of wild reservoirs that maintain the virus in nature. Here we report the antigenic characterization of 254 rabies viruses isolated from 148 nonreservoir and 106 reservoir hosts collected in 27 states of Mexico. Nine out of 11 antigenic variants previously reported in the United States were detected in Mexico by using the limited panel of monoclonal antibodies donated by the Centers for Disease Control and Prevention. Some rabies virus variants were isolated from their natural reservoirs, which were also taxonomically identified. Terrestrial reservoirs included stray dogs with V1, Urocyon cineroargenteus (gray foxes) with V7, and two subspecies of Spilogale putorius (spotted skunks) with different viral variants (V8 and V10). Aerial hosts included Tadarida brasiliensis mexicana and Desmodus rotundus, which harbored V9 and V4 and harbored V11, respectively. All variants, with the exception of V9, were isolated from nonreservoir hosts, while V3, V4, and V5 were not isolated from their natural reservoirs but only from livestock. Rabies virus antigenic typing allowed us to determine rabies reservoirs and their distribution in Mexico, data which will probably improve prevention and control of the illness in humans and in the reservoir hosts. PMID- 11880423 TI - Genetic profiling of enterohemorrhagic Escherichia coli strains in relation to clonality and clinical signs of infection. AB - Sixty-seven human strains of enterohemorrhagic Escherichia coli (EHEC) (from patients with more or less severe symptoms) were serogrouped and arranged according to pulsed-field gel electrophoresis (PFGE) patterns. We used PCR to investigate the strains according to known or putative virulence factors, and associations with disease were studied. All EHEC strains with the same PFGE pattern belonged to the same serogroup. On the contrary, two serogroups (O157 and O8) included strains with different PFGE patterns. We found several different combinations of chromosomal and plasmid-borne determinants, encoding the putative virulence factors, among the strains. As judged from clinical symptoms, there was no marked difference in pathogenicity among the strains and their combinations of virulence traits. All strains of O157 had the genes coding for verocytotoxin (VT) 2, intimin (eaeA), E. coli hemolysin (E-hly), and secreted serine protease (espP). Among EHEC non-O157 strains, the genes coding for VT1 and VT2 were equally dispersed. EaeA positivity was just as common among VT1- as VT2-positive strains. Among the plasmid-borne determinants, E-hly and espP were the most common and E-hly might be a pathogenicity marker among EHEC non-O157 strains. The conclusion is that PFGE is a very useful tool in epidemiological studies. The EHEC plasmids are heterogeneous in their gene composition, with the four plasmid borne determinants found in many combinations. There was no reliable correlation between chromosomal and plasmid-borne virulence factors and human disease. PMID- 11880425 TI - Evaluation of INNO-LIA syphilis assay as a confirmatory test for syphilis. AB - We evaluated the sensitivity and specificity of a new confirmatory test for treponemal antibodies, INNO-LIA Syphilis (Innogenetics NV, Ghent, Belgium), on a large number of sera from a clinical laboratory. This multiparameter line immunoassay (LIA) uses recombinant and synthetic polypeptide antigens derived from Treponema pallidum proteins. In a single-blinded cross-sectional retrospective study, 289 seronegative sera, 219 seropositive sera, and 23 sera with an indeterminate serological status for syphilis were analyzed. All sera were tested by the T. pallidum hemagglutination assay (TPHA), the immunoglobulin (IgG)-fluorescent T. pallidum absorption assay (IgG-FTA-ABS), and the Venereal Disease Research Laboratory (VDRL) test. In addition, some seropositive samples were analyzed by the 19S-IgM-FTA-ABS test, an enzyme immunoassay (IgM-EIA), and the MarDx immunoblotting assay. Based on a consensus diagnosis derived from conventional serology, all of the sera were classified as positive, negative, or indeterminate, and the results were compared with the findings of the INNO-LIA Syphilis assay. The sensitivity and specificity of the LIA were 100% (219 of 219) and 99.3% (286 of 288), respectively. Compared to TPHA and IgG-FTA-ABS, the new test gave a significantly higher number (P = 0.021 and P < 0.0001, respectively) of correct results than either of the other two tests. The multiparameter INNO LIA Syphilis assay is a useful confirmatory test for syphilis because it increases the reliability of syphilis diagnosis with respect to current conventional techniques. PMID- 11880424 TI - Geographic distribution of mating type alleles of Cryptococcus neoformans in four areas of the United States. AB - To better understand the epidemiology and population structure of Cryptococcus neoformans, we determined mating types for 358 C. neoformans strains isolated through the active surveillance program from 1992 to 1994 in four geographic areas in the United States: San Francisco, California; Georgia; Texas; and Alabama. Two assays were used to determine mating types: (i) crossing with standard laboratory tester strains JEC20 and JEC21 on V8 agar medium; and (ii) PCR with the mating type alpha allele-specific primer of the STE12 gene and with serotype (A and D)- and mating type (a and alpha)-specific primers of the STE20 gene. Using these two methods, we found that this sample consisted of the following: (i) 324 serotype A, mating type (MAT) alpha (Aalpha) strains; (ii) 12 serotype D, alpha (Dalpha) strains; (iii) 14 serotype AD strains with mating type alleles Aa and Dalpha (AaDalpha); (iv) 2 serotype AD strains with mating type alleles Aalpha and Da (AalphaDa); (v) 3 serotype B, alpha (Balpha) strains; and (vi) 3 serotype AD strains but with only one mating type allele. No strain with MATa was found within serotype A, B, or D in this collection. Interestingly, 14 of the 19 serotype AD strains contained the Aa allele at the STE20 locus; 13 of these 14 were from San Francisco. Our results suggest that the environment in San Francisco might contain Aa strains capable of mating with Dalpha strains. In addition, our result demonstrate that the sample from San Francisco had a significantly higher proportion of self-fertile strains than those from the other three areas. PMID- 11880426 TI - Genotyping of human papillomavirus in liquid cytology cervical specimens by the PGMY line blot assay and the SPF(10) line probe assay. AB - A comparison of two PCR-based human papillomavirus (HPV) DNA detection and genotyping systems (PGMY LBA and SPF(10) LiPA) was conducted in two laboratories. Both systems are based on broad-spectrum PCR for the detection of HPV DNA, followed by reverse hybridization with type-specific probes. A total of 400 selected cervical scrape specimens in PreservCyt solution (55% normal cytology, 18% atypical squamous cells of unknown significance, 14.8% low-grade squamous intraepithelial lesions [SIL], and 12.5% high-grade SIL) were tested for the presence of HPV DNA. In this selected group of specimens, the overall agreement between the two methods for the detection of any HPV DNA was high (kappa = 0.859). When the 20 common HPV genotypes identified by both methods were considered (HPV types 6, 11, 16, 18, 31, 33, 35, 39, 40, 42, 45, 51, 52, 53, 54, 56, 58, 59, 66, and 68), compatible genotype-specific results were observed in 96.5% of the samples, even when multiple HPV genotypes were present. However, for some specific HPV genotypes, there were significant differences in HPV detection by the two methods. PGMY LBA detected more HPV type 42 (P = 0.002), HPV type 56 (P = 0.039), and HPV type 59 (P < 0.001), whereas SPF(10) LiPA detected more HPV type 31 (P < 0.001) and HPV type 52 (P = 0.031). For the remaining genotypes, including HPV types 16 and 18, the results obtained by the two methods were not significantly different. In general, both genotyping methods are highly suitable for clinical and epidemiological studies. PMID- 11880427 TI - Application of a 5' nuclease assay for detection of Lawsonia intracellularis in fecal samples from pigs. AB - A 5' nuclease assay was developed to detect Lawsonia intracellularis in porcine fecal samples. The specific probe and primers were chosen by using the 16S ribosomal DNA gene as a target. The 5' nuclease assay was used with a total of 204 clinical samples, and the results were compared to those of immunohistochemistry (IM) on ileal sections of the same animals. There was 91% agreement between the results of IM and the 5' nuclease assay. In the 5' nuclease assay, 111 (54%) of the pigs tested positive for L. intracellularis infection, with a mean cycle threshold (Ct) value of 27.2, whereas 98 (48%) of the pigs tested positive by IM. On average, the Ct and DeltaRn values for the positive samples were 27.2 (standard deviation [SD], 3.7) and 1.6 (SD, 0.7), respectively. A Ct value of 27.2 corresponds to a fecal excretion of approximately 10(7) L. intracellularis cells per g of feces. Furthermore, a total of 40 fecal samples derived from a herd known to be free from infection with L. intracellularis all tested negative, with a Ct value of 40. By using a Ct value of 36 as the cutoff limit, the detection limit of the assay was 1 L. intracellularis cell per PCR tube. In conclusion, the 5' nuclease assay that has been developed represents an applicable fast method for detection of L. intracellularis in fecal samples, with a sensitivity and specificity comparable to those of IM. PMID- 11880428 TI - New real-time PCR able to detect in a single tube multiple rifampin resistance mutations and high-level isoniazid resistance mutations in Mycobacterium tuberculosis. AB - The emergence of resistance to antituberculosis drugs is a relevant matter worldwide, but the retrieval of antibiograms for Mycobacterium tuberculosis is severely delayed when phenotypic methods are used. Genotypic methods allow earlier detection of resistance, although conventional approaches are cumbersome or lack sensitivity or specificity. We aimed to design a new real-time PCR method to detect rifampin (RIF)- and isoniazid (INH)-resistant M. tuberculosis strains in a single reaction tube. First, we characterized the resistant isolates in our area of Spain by DNA sequencing. Some mutation was found within the rpoB core region in all the RIF-resistant (RIF(r)) strains. Forty-six percent of the INH resistant (INH(r)) strains showed a mutation in katG codon 315, and most of these were associated with high MICs. Eighteen of the RIF(r), INH(r), and multidrug resistant strains sequenced were tested by our real-time PCR assay; and full concordance of the results of the PCR with the sequencing data was obtained. In addition, a blind test was performed with a panel of 15 different susceptible and resistant strains from throughout Spain, and our results were also in 100% agreement with the sequencing data. Ours is the first assay based on rapid-cycle PCR able to simultaneously detect in a single reaction tube a large variety of mutations associated with RIF resistance (12 different mutations affecting 8 independent codons, including the most prevalent mutations at positions 526 and 531) and the most frequent INH resistance mutations. Our design could be a model for new, rapid genotypic methods able to simultaneously detect a wide variety of antibiotic resistance mutations. PMID- 11880429 TI - Detection of heat-stable antigens of Campylobacter jejuni and C. coli by direct agglutination and passive hemagglutination. AB - The two serotyping schemes for the detection of heat-stable antigens of Campylobacter jejuni and Campylobacter coli use the same strains for antiserum production but differ in the detection systems used for identifying agglutination. The Penner method uses passive hemagglutination (PHA) while the Laboratory of Enteric Pathogens method uses the same antisera but in a whole bacterial-cell direct agglutination (DA) protocol. C. jejuni produces a polysaccharide capsule, which is antigenic, and is the main component detected by the PHA method. The DA method will detect both capsule antigens and lipopolysaccharide (LPS) or lipooligosaccharide (LOS) surface antigens. Comparison of both methods by using a selection of isolates from human infection has shown a range of variation in agglutination specificity, reflecting the differences in antigens detected by the two methods. While 27.4% of the 416 C. jejuni isolates reacted with the antisera raised against the same type strains by either method, the majority showed a range of more complex relationships. None of the 37 C. coli isolates reacted with the same antiserum by both methods. Together the two schemes gave a total of 102 distinct combined serogroups for C. jejuni and 16 for C. coli. Thus, while some clonally related isolates share the same capsule and LOS or LPS antigens, other strains appear to have a common capsule antigen but differ in their LPS or LOS structures or vice versa. PMID- 11880430 TI - Molecular analysis of bacterial species associated with childhood caries. AB - Although substantial epidemiologic evidence links Streptococcus mutans to caries, the pathobiology of caries may involve more complex communities of bacterial species. Molecular methods for bacterial identification and enumeration now make it possible to more precisely study the microbiota associated with dental caries. The purpose of this study was to compare the bacteria found in early childhood caries (ECC) to those found in caries-free children by using molecular identification methods. Cloning and sequencing of bacterial 16S ribosomal DNAs from a healthy subject and a subject with ECC were used for identification of novel species or uncultivated phylotypes and species not previously associated with dental caries. Ten novel phylotypes were identified. A number of species or phylotypes that may play a role in health or disease were identified and warrant further investigation. In addition, quantitative measurements for 23 previously known bacterial species or species groups were obtained by a reverse capture checkerboard assay for 30 subjects with caries and 30 healthy controls. Significant differences were observed for nine species: S. sanguinis was associated with health and, in order of decreasing cell numbers, Actinomyces gerencseriae, Bifidobacterium, S. mutans, Veillonella, S. salivarius, S. constellatus, S. parasanguinis, and Lactobacillus fermentum were associated with caries. These data suggest that A. gerencseriae and other Actinomyces species may play an important role in caries initiation and that a novel Bifidobacterium may be a major pathogen in deep caries. Further investigation could lead to the identification of targets for biological interventions in the caries process and thereby contribute to improved prevention of and treatment for this significant public health problem. PMID- 11880431 TI - Development and evaluation of a DNA enzyme immunoassay method for env genotyping of subtypes A through G of human immunodeficiency virus type 1 group M, with discrimination of the circulating recombinant forms CRF01_AE and CRF02_AG. AB - The tools currently available for genetic subtyping of human immunodeficiency virus type 1 are laborious or can be used only for the analysis of a limited number of samples and/or subtypes. We developed and evaluated a molecular biology based method using subtype-specific oligonucleotide probes for env genotyping of subtypes A through G, CRF01_AE, and CRF02_AG. DNA enzyme immunoassay (DEIA) genotyping is based on nested PCR amplification of the 5' end of the env gene (proviral DNA), followed by subtype-specific hybridization and immunoenzymatic detection on microplates. DEIA genotyping was validated with a large number of samples (n = 128) collected in Europe (France; n = 47), West-Central Africa (Cameroon; n = 36), and West Africa (Senegal; n = 45). Three different formats, depending on the distribution of subtypes in the three countries, were developed. The results were compared with those obtained by sequencing of the V3-V5 region and phylogenetic analysis or an env heteroduplex mobility assay. Additional sequencing and phylogenetic analyses of the DEIA region (the first codon of the env coding sequence to the middle of conserved region C1 of gp120) were performed to investigate the reasons for discrepancies. Intense and highly specific reactions between the oligonucleotide probes and the corresponding samples were observed. Overall, correct identification was achieved for 107 of 128 samples (83.6%). One sample was not amplified, 10 (8%) were nontypeable (NT), and 10 (8%) were misidentified. Six of the 10 discordant samples were further investigated by phylogenetic analysis, which indicated that these samples corresponded to recombinants involving the env 5' end and the V3 and V5 regions of the two parental clades. Sequencing of NT samples showed numerous differences between sample and probe sequences, resulting in a lack of hybridization, and revealed the limitations of the selected probes in terms of specificity and sensitivity. We demonstrated the feasibility of DEIA genotyping: six subtypes plus the two most prevalent circulating recombinant forms were discriminated by using the 5' end of the env gene. This method can be adapted to the local situation by including only probes that correspond to the prevalent strains. PMID- 11880432 TI - Genomic variation of Bartonella henselae strains detected in lymph nodes of patients with cat scratch disease. AB - Bartonella henselae is the primary agent of cat scratch disease (CSD). In order to study the genetic variation of B. henselae and the correlation of the various genotypes with epidemiological and clinical findings, two seminested, groEL- and pap31-based PCR assays were carried out with specimens from 273 patients. Amplicons were sequenced to determine the genotype of the causative Bartonella species. Compared to our reference intergenic spacer region-based PCR, the groEL- and pap31-based assays were 1.7 and 1.9 times more sensitive, respectively. All 107 positive patients were infected with B. henselae; neither Bartonella clarridgeiae nor other species were detected. Based on the groEL and pap31 sequences, B. henselae amplicons were classified into two genogroups, Marseille and Houston-1, and into four variants, Marseille, CAL-1, Houston-1, and a new variant, ZF-1. Patients infected with either one or the other genogroup did not exhibit different epidemiological or clinical characteristics. Our study highlights the genotypic heterogeneity of B. henselae in patients with CSD. PMID- 11880433 TI - Environmental occurrence of Madurella mycetomatis, the major agent of human eumycetoma in Sudan. AB - Madurella mycetomatis is the main causative agent of human eumycetoma, a severe debilitating disease endemic in Sudan. It has been suggested that eumycetoma has a soil-borne or thorn prick-mediated origin. For this reason, efforts were undertaken to culture M. mycetomatis from soil samples (n = 43) and thorn collections (n = 35) derived from areas in which it is endemic. However, ribosomal sequencing data revealed that the black fungi obtained all belonged to other fungal species. In addition, we performed PCR-mediated detection followed by restriction fragment length polymorphism (RFLP) analysis for the identification of M. mycetomatis DNA from the environmental samples as well as biopsies from patients with mycetoma. In the case of the Sudanese soil samples, 17 out of 74 (23%) samples were positive for M. mycetomatis DNA. Among the thorn collections, 1 out of 22 (5%) was positive in the PCR. All PCR RFLP patterns clearly indicated the presence of M. mycetomatis. In contrast, 15 Dutch and English control soil samples were all negative. Clinically and environmentally obtained fungal PCR products share the same PCR RFLP patterns, suggesting identity, at least at the species level. These observations support the hypothesis that eumycetoma is primarily environmentally acquired and suggest that M. mycetomatis needs special conditions for growth, as direct isolation from the environment seems to be impossible. PMID- 11880434 TI - Enzyme-linked immunosorbent assay based on soluble promastigote antigen detects immunoglobulin M (IgM) and IgG antibodies in sera from cases of visceral and cutaneous leishmaniasis. AB - Leishmaniasis causes significant morbidity and mortality in areas where it is endemic. In areas where it is nonendemic, global travel and increased incidence of the disease in human immunodeficiency virus and intravenous-drug user populations are also causes for concern. The unavailability of rapid and reliable tests for diagnosis of the various leishmaniases makes patient management difficult. We have developed an enzyme-linked immunosorbent assay (ELISA) that can detect immunoglobulin M (IgM) and IgG antibodies in patients with visceral and cutaneous leishmaniasis. These practical assays are based on soluble antigens from promastigotes cultivated in a protein-free medium. In preliminary studies, 129 visceral (Brazil, Italy, North Africa, and Nepal) and 143 cutaneous (Brazil) leishmaniasis patients with controls were tested. Overall, the tests showed a sensitivity of 95.1%. In addition, the ELISA correctly identified 42 sera from Brazilian dogs with canine leishmaniasis and 10 healthy controls. Serological tests for the various clinical manifestations of leishmaniasis could be useful epidemiological and patient management tools in populations of areas of endemicity and nonendemicity. PMID- 11880435 TI - Periodontal bacteria in rabbit mandibular and maxillary abscesses. AB - Despite the high incidence of odontogenic abscesses in pet rabbits, published data on the bacteriology of these infections are lacking, and clinical cultures are often ambiguous, making antibiotic choices difficult. In order to define the bacteriology of these infections, 12 rabbit mandibular and maxillary abscesses were cultured aerobically and anaerobically. All specimens yielded pathogenic bacteria, including Fusobacterium nucleatum, Prevotella heparinolytica, Prevotella spp., Peptostreptococcus micros, Streptococcus milleri group, Actinomyces israelii, and Arcanobacterium haemolyticum. These organisms are consistent with the characterized bacteriology of periodontal disease in human and other mammalian studies. The isolates were tested against 10 antimicrobial agents commonly used to treat rabbits; 100% of the strains tested were susceptible to clindamycin, 96% were susceptible to penicillin and ceftriaxone, 54% were susceptible to ciprofloxacin, and only 7% were susceptible to trimethoprim-sulfamethoxazole. PMID- 11880436 TI - Nine-year surveillance of methicillin-resistant Staphylococcus aureus in a hospital suggests instability of mecA DNA region in an epidemic strain. AB - The distributions of the antibiotic resistance patterns in a population of Staphylococcus aureus isolates from a teaching hospital were studied over a 9 year period. The results indicate the existence of successive major epidemic methicillin-resistant strains and the emergence of a methicillin-susceptible strain with an unusual resistance pattern. Our findings suggest that this methicillin-susceptible S. aureus strain could be derived from the dominant gentamicin-susceptible methicillin-resistant S. aureus strain with the loss of a 40-kb DNA fragment. PMID- 11880437 TI - Fatal case of Campylobacter lari prosthetic joint infection and bacteremia in an immunocompetent patient. AB - Campylobacter lari is an infrequent cause of intestinal and extraintestinal infection in humans. We report a case of C. lari prosthetic joint infection and bacteremia in an 81-year-old immunocompetent man. The infection was associated with septic shock and fatal outcome. C. lari may cause severe disease, even in an immunocompetent host. PMID- 11880438 TI - PCR detection and DNA sequence analysis of the regulatory region of lymphotropic papovavirus in peripheral blood mononuclear cells of an immunocompromised rhesus macaque. AB - A lymphotropic papovavirus (LPV) archetypal regulatory region was amplified from DNA from the blood of an immunocompromised rhesus monkey. We believe this is the first nonserological evidence of LPV infection in rhesus monkeys. PMID- 11880439 TI - Rapid detection of herpes simplex virus DNA in genital ulcers by real-time PCR using SYBR green I dye as the detection signal. AB - We have evaluated a real-time PCR procedure based on the LightCycler technology for rapid detection of herpes simplex virus (HSV) in genital lesions. Two sets of primers, corresponding to the thymidine kinase and DNA polymerase regions, were used for the amplification reactions in separate capillaries containing the SYBR Green I dye as detection signal. In 28 of 118 samples (24%), HSV was isolated by conventional cell culture. All cell culture-positive samples were also positive by real-time PCR. Six additional cell culture-negative samples were positive by PCR with both sets of primers. Total processing time was less than 3 h. Real-time PCR using SYBR Green I as detection signal is a sensitive procedure for the rapid diagnosis of HSV in genital lesions. PMID- 11880440 TI - Antimicrobial surveillance of Haemophilus influenzae in the United States during 2000-2001 leads to detection of clonal dissemination of a beta-lactamase-negative and ampicillin-resistant strain. AB - A 2000-2001 U.S. Haemophilus influenzae surveillance study (n = 1,434) detected nine (0.6%) beta-lactamase-negative and ampicillin-resistant (BLNAR) isolates collected from two different hospitals. The MICs of ampicillin for all nine isolates were 4 microg/ml, with results being reproducible; and all nine isolates were susceptible to amoxicillin-clavulanate, cefuroxime, cefprozil, macrolides, and fluoroquinolones. Pulsed-field gel electrophoresis of genomic DNA following SmaI digestion demonstrated identical patterns for each of the nine isolates, suggesting intra- and interhospital dissemination of a BLNAR clone. PMID- 11880441 TI - Ratio of two successive optical densities from the Roche HIV-1 monitor test as a measure of accuracy of estimates of human immunodeficiency virus RNA concentration. AB - In the Roche Amplicor HIV-1 Monitor Test, the ratio of the optical density used to estimate the RNA concentration and the optical density of the preceding or the following dilution can be used to identify specimens with inaccurate results which should be retested. We present an algorithm for the identification of such results. PMID- 11880442 TI - Comparison of molecular and conventional methods for typing of enteroviral isolates. AB - Twenty-eight enteroviral isolates obtained from various clinical specimens were typed by Lim-Benyesh-Melnick (LBM) pool-based neutralization, PCR-restriction fragment length polymorphism (RFLP), and partial sequencing of the VP1 region of the enteroviral genome. Sequencing was found to be a good alternative to LBM typing, while PCR-RFLP was inadequate for identification of enteroviral isolates. PMID- 11880443 TI - Comparative performance of herpes simplex virus type 1-specific serologic assays from MRL and Meridian Diagnostics. AB - Two companies, MRL and Meridian Diagnostics, have developed Food and Drug Administration-approved herpes simplex virus type 1 type-specific enzyme immunoassays. The sensitivity, specificity, and overall testing efficiency of these assays were 98.2, 93.8, and 96.6% for MRL and 98.8, 99.0, and 98.1% for Meridian, making both of these kits suitable for use in the clinical lab. PMID- 11880444 TI - Genetic characterization of H1N2 influenza A viruses isolated from pigs throughout the United States. AB - An H1N2 influenza A virus was isolated from a pig in the United States for the first time in 1999 (A. I. Karasin, G. A. Anderson, and C. W. Olsen, J. Clin. Microbiol. 38:2453-2456, 2000). H1N2 viruses have been isolated subsequently from pigs in many states. Phylogenetic analyses of eight such viruses isolated from pigs in Indiana, Illinois, Minnesota, Ohio, Iowa, and North Carolina during 2000 to 2001 showed that these viruses are all of the same reassortant genotype as that of the initial H1N2 isolate from 1999. PMID- 11880445 TI - Monoclonal antibodies specific for hippurate hydrolase of Campylobacter jejuni. AB - Eleven monoclonal antibodies raised against recombinant Campylobacter jejuni hippurate hydrolase were tested for binding to lysates from 19 C. jejuni strains, 12 other Campylobacter strains, and 21 non-Campylobacter strains. Several monoclonal antibodies bound to C. jejuni but not to other Campylobacter species and may be useful in a species-specific immunoassay. PMID- 11880446 TI - Neisseria meningitidis serogroups W135 and A were equally prevalent among meningitis cases occurring at the end of the 2001 epidemics in Burkina Faso and Niger. AB - Meningococcal infections occur as epidemics in the African meningitis belt. Neisseria meningitidis serogroup A is predominantly involved in these epidemics. We report here new data on the involvement of both serogroups A and W135 in meningitis cases in Burkina Faso and Niger at the end of the 2001 epidemic. PMID- 11880447 TI - Biochemical and susceptibility tests useful for identification of nonfermenting gram-negative rods. AB - Six hundred nineteen strains of nonfermenting gram-negative rods were tested for alkaline phosphatase, benzyl-arginine arylamidase, pyrrolidonyl arylamidase, ethylene glycol acidification, and susceptibility to desferrioxamine and colistin. The results were highly discriminant. Therefore, the proposed tests may be helpful for the identification of this group of organisms. PMID- 11880448 TI - Results of human papillomavirus DNA testing with the hybrid capture 2 assay are reproducible. AB - Reproducibility of the Hybrid Capture 2 Test (HC 2) for human papillomavirus (HPV) DNA detection was evaluated by assaying frozen cervical specimens in 1997 and again in 2001 from 1,775 women with normal cervical cytology. Using a cutoff point of 1.0 pg of HPV DNA/ml between a negative and a positive test result, the result of the kappa test for agreement was 0.72 (a kappa value of >0.60 is considered good agreement). Using cutoff points of 1.0 and 10.0 pg/ml between negative and low positive and between low positive and high positive, respectively, the kappa was 0.68 and the linear-weighted kappa was 0.76. The results of this study indicate that HC 2 testing is reproducible even among cytologically normal women with low test values. PMID- 11880449 TI - rpoB genotypes of Mycobacterium tuberculosis Beijing family isolates from East Asian countries. AB - The 81-bp region of the rpoB gene in 66 Rif(r) Mycobacterium tuberculosis isolates from China, Japan, Korea, and Taiwan was analyzed. Twelve single nucleotide substitutions in the rpoB gene were detected. The most prevalent mutations were at Ser-531 (52%), Asp-516 (17%), and His-526 (11%). Mutations were not found in seven (11%) of the isolates. Higher mutation rates in 50 Beijing family isolates were found than in other isolates for mutations at Asp-516 (18 and 12.5%, respectively) and His-526 (12 and 6.3%, respectively). The different rates of mutation may reflect the choice of rifamycin analogs. PMID- 11880450 TI - Two different extended-spectrum beta-lactamases (ESBLs) in one of the first ESBL producing salmonella isolates in Poland. AB - Two extended-spectrum beta-lactamase (ESBL)-producing salmonella isolates, Salmonella enterica serovar Enteritidis and Salmonella enterica serovar Typhimurium, were analyzed. Both isolates produced the CTX-M-3 ESBL; however, their bla(CTX-M-3) genes were located on different plasmids. The serovar Typhimurium isolate also expressed another ESBL, SHV-2a, and probably the two ESBL genes had been acquired independently by the strain. PMID- 11880451 TI - Fatal pulmonary Nocardia farcinica infection. AB - Nocardia farcinica infections are rare and potentially life threatening. Identification is based on growth at 45 degreesC, opacification of Middlebrook 7H10 agar, and resistance to antibiotics. We describe a case of fatal pulmonary N. farcinica infection in a patient with pneumoconiosis that was diagnosed by culture of sputum onto selective media. PMID- 11880452 TI - Successful outcome of treatment of a disseminated infection due to Fusarium dimerum in a leukemia patient. AB - We report the first case of proven disseminated infection due to Fusarium dimerum associated with a favorable outcome in a patient with acute leukemia and prolonged neutropenia. The patient presented persistent fever, multiple necrotic skin lesions, and bilateral pneumopathy. F. dimerum was first isolated from three blood cultures and then from a skin biopsy and a mouth wash. Microscopy of positive blood cultures showed hyphae with phialides and few curved unicellular and some rare bicellular phialoconidia, permitting immediate presumptive identification of the genus FUSARIUM: The patient failed to respond to conventional amphotericin B but recovered after treatment was switched to amphotericin B-lipid complex and neutrophil recovery. PMID- 11880453 TI - Molecular epidemiology of Mycobacterium malmoense infections in Scotland. AB - Clinical isolates of Mycobacterium malmoense collected over 5 years from patients across Scotland with a variety of diseases have been characterized by pulsed field gel electrophoresis (PFGE), ribotyping, and 16S ribosomal DNA gene sequencing. Results indicate that this species harbors little genetic diversity and that the different strain types that were identified by PFGE showed no correlation with geographical origin or date of isolation. PMID- 11880454 TI - Unusual outbreak of clinical mastitis in dairy sheep caused by Streptococcus equi subsp. zooepidemicus. AB - This work describes an outbreak of clinical mastitis affecting 13 of 58 lactating ewes due to Streptococcus equi subsp. zooepidemicus. S. equi subsp. zooepidemicus was isolated in pure culture from all milk samples. All the clinical isolates had identical biochemical profiles and antimicrobial susceptibility patterns and also exhibited indistinguishable macrorestriction patterns by pulsed-field gel electrophoresis, indicating that all cases of mastitis were produced by a single strain. PMID- 11880455 TI - Staphylococcus heterogeneously resistant to vancomycin in China and antimicrobial activities of imipenem and vancomycin in combination against it. AB - Of 115 methicillin-resistant Staphylococcus strains collected from sputum specimens, 34 strains reduced susceptibility to vancomycin, 9 of which emerged as heterogeneous vancomycin-resistant strains (hetero-VRS), with various degrees of vancomycin resistance at a frequency of 10(-6) or higher. Seventy-six percent (19 of 25) of non-hetero-VRS and 100% (9 of 9) of hetero-VRS were susceptible to synergistic treatment with vancomycin and imipenem. Clinical clearance between 9 hetero-VRS and 25 non-hetero-VRS had an obvious statistical significance (P = 0.001). The hetero-VRS may play an important role in vancomycin therapy failure. PMID- 11880456 TI - Pulmonary colonization by Chrysosporium zonatum associated with allergic inflammation in an immunocompetent subject. AB - We report a case of noninvasive pulmonary disease due to Chrysosporium zonatum in an immunocompetent male. The fungus colonized an existing tuberculous cavity and was isolated from transbronchial lavage fluid and from a percutaneous aspiration specimen. The disease was accompanied by the unusual feature of an allergic reaction. The fungus ball was successfully treated by intracavitary administration of amphotericin B. C. zonatum is the anamorph of the heterothallic ascomycete Uncinocarpus orissi, and the identity of the case isolate was verified by formation of ascospores in mating tests with reference isolates. PMID- 11880457 TI - First isolation of reddish-pigmented Candida (Torulopsis) glabrata from a clinical specimen. PMID- 11880458 TI - Quantitative detection of Tropheryma whipplei DNA by real-time PCR. PMID- 11880459 TI - Helicobacter infection and spontaneous bacterial peritonitis. PMID- 11880460 TI - Crimean-Congo hemorrhagic fever in Kosovo. PMID- 11880461 TI - Novel PCR-probe assay for detection of and discrimination between Legionella pneumophila and other Legionella species in clinical samples. PMID- 11880462 TI - Qiagen DNA extraction kits for sample preparation for legionella PCR are not suitable for diagnostic purposes. PMID- 11880463 TI - Capsular Vi polysaccharide antigen in Salmonella enterica serovar typhi isolates. PMID- 11880464 TI - Ras and Seppuku in neuroblastoma. PMID- 11880465 TI - Does practice make perfect when interpreting mammography? PMID- 11880466 TI - Battle over generic taxol concludes, but controversy continues. PMID- 11880468 TI - Biotech firm faces challenges from FDA, falling stock prices. PMID- 11880470 TI - Direct-to-consumer marketing: how has it fared? PMID- 11880471 TI - AstraZeneca reprimanded for nolvadex advertisement. PMID- 11880472 TI - Stool test identifies markers for colorectal cancer, studies show. PMID- 11880473 TI - Racial and ethnic disparities in the receipt of cancer treatment. AB - A disproportionate number of cancer deaths occur among racial/ethnic minorities, particularly African Americans, who have a 33% higher risk of dying of cancer than whites. Although differences in incidence and stage of disease at diagnosis may contribute to racial disparities in mortality, evidence of racial disparities in the receipt of treatment of other chronic diseases raises questions about the possible role of inequities in the receipt of cancer treatment. To evaluate racial/ethnic disparities in the receipt of cancer treatment, we examined the published literature that addressed access/use of specific cancer treatment procedures, trends in patterns of use, or survival studies. We found evidence of racial disparities in receipt of definitive primary therapy, conservative therapy, and adjuvant therapy. These treatment differences could not be completely explained by racial/ethnic variation in clinically relevant factors. In many studies, these treatment differences were associated with an adverse impact on the health outcomes of racial/ethnic minorities, including more frequent recurrence, shorter disease-free survival, and higher mortality. Reducing the influence of nonclinical factors on the receipt of cancer treatment may, therefore, provide an important means of reducing racial/ethnic disparities in health. New data resources and improved study methodology are needed to better identify and quantify the full spectrum of nonclinical factors that contribute to the higher cancer mortality among racial/ethnic minorities and to develop strategies to facilitate receipt of appropriate cancer care for all patients. PMID- 11880474 TI - Increased Ras expression and caspase-independent neuroblastoma cell death: possible mechanism of spontaneous neuroblastoma regression. AB - BACKGROUND: Neuroblastoma undergoes spontaneous regression frequently during its natural course. Although programmed cell death (PCD) has been implicated in this process, accumulating evidence suggests that apoptosis, a form of PCD that is regulated by caspases, may not play a major role. We examined the mechanism(s) of spontaneous regression of neuroblastoma, focusing on the role of Ras, a favorable prognostic marker of neuroblastoma. METHODS: Tumor tissues were analyzed by light microscopy, electron microscopy, and immunohistochemistry to examine cell degeneration and expression of Ras and several indicators of PCD. Cell degeneration was also studied in vitro in neuroblastoma cells transfected with the H-ras gene. All statistical tests were two-sided. RESULTS: Immunohistochemical analyses revealed that Ras expression was increased in areas of cellular degeneration lacking apoptotic characteristics. The degenerating cells were fragmented without nuclear condensation and, essentially, lacked caspase-3 activation and apoptotic DNA fragmentation. These cells had ultrastructural features of autophagic degeneration, another form of PCD that is distinct from apoptosis. Focal areas of degeneration associated with Ras expression were seen more frequently in tumors from patients detected in a mass screening program (53 [60.9%] of 87) than in tumors from clinically detected, advanced-stage patients over 1 year of age (7 [29.2%] of 24) (P =.006; chi-square test), suggesting a positive relationship between Ras-associated degeneration and probability of spontaneous regression/favorable prognosis. The characteristic features of Ras-associated nonapoptotic degeneration observed in tumor samples were recapitulated in vitro by transfection-mediated Ras expression, and Ras mediated degeneration was augmented by TrkA, another favorable prognostic marker. CONCLUSIONS: High-level expression of H-Ras in neuroblastoma cells is associated with caspase cascade-independent, nonapoptotic PCD. This Ras-mediated nonapoptotic tumor cell death may play a key role in spontaneous regression of neuroblastoma. PMID- 11880475 TI - Improving the accuracy of mammography: volume and outcome relationships. AB - BACKGROUND: Countries with centralized, high-volume mammography screening programs, such as the U.K. and Sweden, emphasize high specificity (low percentage of false positives) and high sensitivity (high percentage of true positives). By contrast, the United States does not have centralized, high-volume screening programs, emphasizes high sensitivity, and has lower average specificity. We investigated whether high sensitivity can be achieved in the context of high specificity and whether the number of mammograms read per radiologist (reader volume) drives both sensitivity and specificity. METHODS: The U.K.'s National Health Service Breast Screening Programme uses the PERFORMS 2 test as a teaching and assessment tool for radiologists. The same 60-film PERFORMS 2 test was given to 194 high-volume U.K. radiologists and to 60 U.S. radiologists, who were assigned to low-, medium-, or high-volume groups on the basis of the number of mammograms read per month. The standard binormal receiver-operating characteristic (ROC) model was fitted to the data of individual readers. Detection accuracy was measured by the sensitivity at specificity = 0.90, and differences among sensitivities were determined by analysis of variance. RESULTS: The average sensitivity at specificity = 0.90 was 0.785 for U.K. radiologists, 0.756 for high-volume U.S. radiologists, 0.702 for medium-volume U.S. radiologists, and 0.648 for low-volume U.S. radiologists. At this specificity, low-volume U.S. radiologists had statistically significantly lower sensitivity than either high-volume U.S. radiologists or U.K. radiologists, and medium-volume U.S. radiologists had statistically significantly lower sensitivity than U.K. radiologists (P<.001, for all comparisons). CONCLUSIONS: Reader volume is an important determinant of mammogram sensitivity and specificity. High sensitivity (high cancer detection rate) can be achieved with high specificity (low false positive rate) in high-volume centers. This study suggests that there is great potential for optimizing mammography screening. PMID- 11880476 TI - Gastrin-releasing peptide receptor-mediated autocrine growth in squamous cell carcinoma of the head and neck. AB - BACKGROUND: Gastrin-releasing peptide receptor (GRPR)-mediated autocrine growth appears to be an early marker of susceptibility to tobacco-related lung cancers. Because expression of GRPR, however, has not been reported in squamous cell carcinoma of the head and neck (SCCHN), we investigated its expression and that of its ligand GRP in normal mucosa and SCCHN tissues and the involvement of these proteins in the proliferation of SCCHN cells. METHODS: We assessed GRPR messenger RNA (mRNA) expression in specimens from 25 patients with SCCHN, six control noncancer patients, and 14 SCCHN cell lines by use of quantitative reverse transcriptase-polymerase chain reaction. We used neutralizing GRP monoclonal antibody 2A11 to block the GRP-GRPR interaction in SCCHN cell lines and xenografts and assessed the antibody's effect on proliferation by counting cultured cells or measuring xenograft tumor volume in vivo. All statistical tests were two-sided. RESULTS: Tumor and mucosa tissues, respectively, from SCCHN patients expressed sixfold and fourfold higher levels of GRPR mRNA than normal mucosa tissue from noncancer patients (P<.001). The levels of GRPR expression in the tumor and adjacent normal epithelium of individual patients with SCCHN were correlated (r =.652; P =.001), suggesting that increased GRPR expression is an early event in SCCHN formation. SCCHN cells expressed fivefold higher levels of GRPR mRNA than did cultured normal mucosal epithelial cells (P =.005). GRP stimulated proliferation of SCCHN cells in a dose-dependent fashion (P =.006). Neutralizing GRP monoclonal antibody 2A11 inhibited SCCHN cell proliferation in vitro and in vivo. Median survival was 54 months in patients with higher levels of GRPR mRNA and was not reached in those with lower levels. CONCLUSIONS: GRP and GRPR appear to participate in an autocrine regulatory pathway in SCCHN. Thus, strategies that specifically target GRP and/or GRPR may be effective therapeutic approaches for this disease. PMID- 11880477 TI - Methylation and inactivation of estrogen, progesterone, and androgen receptors in prostate cancer. AB - BACKGROUND: Prostate cancer development is initially steroid hormone dependent. Estrogen receptors (ERs), androgen receptors (ARs), and progesterone receptors (PRs) have been identified in normal and cancerous prostate tissues. We investigated whether the promoter regions of these steroid receptor genes are methylated and inactivated in prostate cancer cells and tissues. METHODS: The expression and promoter methylation status of three ERalpha isoforms (ERalpha-A, ERalpha-B, and ERalpha-C), ERbeta, two PR isoforms (PR-A and PR-B), and AR were investigated in five prostate cancer cell lines (ND1, DU145, PC3, LNCaP, and DUPro) and in pairs of normal and cancerous prostate tissues from 38 patients with prostate cancer. Methylation-specific polymerase chain reaction, reverse transcription--polymerase chain reaction, and 5' rapid amplification of complementary DNA ends were used. All statistical tests were two-sided. RESULTS: ERalpha-C was expressed in all cell lines, but ERalpha-A and ERalpha-B were not expressed in any cell line. ERalpha-A and ERalpha-B promoters were methylated, but ERalpha-C was unmethylated. Promoters for ERbeta, AR, PR-A, and PR-B were methylated and thus inactivated in some cell lines but not in others. Treating cells with the demethylating reagent 5-aza-2'-deoxycytidine restored expression of all steroid receptor genes with previously methylated promoters. All 38 pairs of cancer and normal tissues had unmethylated ERalpha-C promoters. Thirty-six (95%) of 38 cancers had methylated ERalpha-A, 35 (92%) of 38 cancers had methylated ERalpha-B, but all normal tissues had unmethylated ERalpha-A and ERalpha-B (both P<.001). ERbeta was methylated in 30 (79%) of 38 cancers but unmethylated in all normal tissues. AR was methylated in three (8%) of 38 cancers but unmethylated in all normal tissues. PR-A and PR-B were unmethylated in all tissues. CONCLUSION: Certain steroid receptor genes appear to be inactivated by CpG methylation in prostate cancer tissue and cell lines. PMID- 11880478 TI - A prospective study of tomato products, lycopene, and prostate cancer risk. AB - BACKGROUND: Some data, including our findings from the Health Professionals Follow-Up Study (HPFS) from 1986 through January 31, 1992, suggest that frequent intake of tomato products or lycopene, a carotenoid from tomatoes, is associated with reduced risk of prostate cancer. Overall, however, the data are inconclusive. We evaluated additional data from the HPFS to determine if the association would persist. METHODS: We ascertained prostate cancer cases from 1986 through January 31, 1998, among 47 365 HPFS participants who completed dietary questionnaires in 1986, 1990, and 1994. We used pooled logistic regression to compute multivariate relative risks (RR) and 95% confidence intervals (CIs). All statistical tests were two-sided. RESULTS: From 1986 through January 31, 1998, 2481 men in the study developed prostate cancer. Results for the period from 1992 through 1998 confirmed our previous findings---that frequent tomato or lycopene intake was associated with a reduced risk of prostate cancer. Similarly, for the entire period of 1986 through 1998, using the cumulative average of the three dietary questionnaires, lycopene intake was associated with reduced risk of prostate cancer (RR for high versus low quintiles = 0.84; 95% CI = 0.73 to 0.96; P(trend) =.003); intake of tomato sauce, the primary source of bioavailable lycopene, was associated with an even greater reduction in prostate cancer risk (RR for 2+ servings/week versus <1 serving/month = 0.77; 95% CI = 0.66 to 0.90; P(trend)<.001), especially for extraprostatic cancers (RR = 0.65; 95% CI = 0.42 to 0.99). These associations persisted in analyses controlling for fruit and vegetable consumption and for olive oil use (a marker for Mediterranean diet) and were observed separately in men of Southern European or other Caucasian ancestry. CONCLUSION: Frequent consumption of tomato products is associated with a lower risk of prostate cancer. The magnitude of the association was moderate enough that it could be missed in a small study or one with substantial errors in measurement or based on a single dietary assessment. PMID- 11880479 TI - Supporting databases for neuroscience research. PMID- 11880480 TI - Dendritic calcium encodes striatal neuron output during up-states. AB - Striatal spiny projection neurons control basal ganglia outputs via action potential bursts conveyed to the globus pallidus and substantia nigra. Accordingly, burst activity in these neurons contributes importantly to basal ganglia function and dysfunction. These bursts are driven by multiple corticostriatal inputs that depolarize spiny projection neurons from their resting potential of approximately -85 mV, which is the down-state, to a subthreshold up-state of -55 mV. To understand dendritic processing of bursts during up-states, changes in intracellular calcium concentration ([Ca2+]i) were measured in striatal spiny projection neurons from cortex-striatum-substantia nigra organotypic cultures grown for 5-6 weeks using somatic whole-cell patch recording and Fura-2. During up-states, [Ca2+]i transients at soma and primary, secondary, and tertiary dendrites were highly correlated with burst strength (i.e., the number of spontaneous action potentials). During down-states, the action potentials evoked by somatic current pulses elicited [Ca2+]i transients in higher-order dendrites that were also correlated with burst strength. Evoked bursts during up-states increased dendritic [Ca2+]i transients supralinearly by >200% compared with the down-state. In the presence of tetrodotoxin, burst-like voltage commands failed to elicit [Ca2+]i transients at higher-order dendrites. Thus, dendritic [Ca2+]i transients in spiny projection neurons encode somatic bursts supralinearly during up-states through active propagation of action potentials along dendrites. We suggest that this conveys information about the contribution of a spiny projection neuron to a basal ganglia output specifically back to the corticostriatal synapses involved in generating these outputs. PMID- 11880481 TI - Activation by serotonin and noradrenaline of vasopressin and oxytocin expression in the mouse paraventricular and supraoptic nuclei. AB - Noradrenaline and serotonin are known to control arginine-vasopressin (AVP) and oxytocin (OT) secretion in the systemic circulation. The aim of the current study was to investigate whether these monoamines are also able to influence AVP and OT expression in the paraventricular (PVN) and supraoptic nuclei (SON). To test this hypothesis, we used the Tg8 transgenic mice KO for the monoamine oxidase-A gene, which present high levels of noradrenaline and serotonin in the brain. AVP and OT expression were evaluated at peptide and mRNA levels by immunohistochemistry, enzyme immunoassay, and in situ hybridization. Compared with wild type, the amounts of AVP, OT, AVP mRNA, and OT mRNA were increased in the PVN and SON in Tg8 mice. To distinguish the respective contributions of noradrenaline and serotonin to these modifications, we treated Tg8 mice with a synthesis inhibitor of either catecholamines [alpha-methylparatyrosine (alpha-MPT)] or serotonin [parachlorophenylalanine (pCPA)]. Administration of alpha-MPT to Tg8 mice induced a decline in the amounts of AVP, OT, and their mRNA in the PVN and SON. The pCPA treatment in Tg8 mice was also associated with a decrease in OT expression in the PVN and SON and in AVP expression in the PVN, but not in the SON. These results suggest that noradrenaline may activate AVP and OT expression in the PVN and SON. Likewise, serotonin is proposed to stimulate AVP and OT expression in the PVN and only OT expression in the SON. PMID- 11880482 TI - Astroglial contribution to brain energy metabolism in humans revealed by 13C nuclear magnetic resonance spectroscopy: elucidation of the dominant pathway for neurotransmitter glutamate repletion and measurement of astrocytic oxidative metabolism. AB - Increasing evidence supports a crucial role for glial metabolism in maintaining proper synaptic function and in the etiology of neurological disease. However, the study of glial metabolism in humans has been hampered by the lack of noninvasive methods. To specifically measure the contribution of astroglia to brain energy metabolism in humans, we used a novel noninvasive nuclear magnetic resonance spectroscopic approach. We measured carbon 13 incorporation into brain glutamate and glutamine in eight volunteers during an intravenous infusion of [2 13C] acetate, which has been shown in animal models to be metabolized specifically in astroglia. Mathematical modeling of the three established pathways for neurotransmitter glutamate repletion indicates that the glutamate/glutamine neurotransmitter cycle between astroglia and neurons (0.32 +/ 0.07 micromol x gm(-1) x min(-1)) is the major pathway for neuronal glutamate repletion and that the astroglial TCA cycle flux (0.14 +/- 0.06 micromol x gm(-1) x min(-1)) accounts for approximately 14% of brain oxygen consumption. Up to 30% of the glutamine transferred to the neurons by the cycle may derive from replacement of oxidized glutamate by anaplerosis. The further application of this approach could potentially enlighten the role of astroglia in supporting brain glutamatergic activity and in neurological and psychiatric disease. PMID- 11880483 TI - Brain-derived neurotrophic factor induces long-term potentiation in intact adult hippocampus: requirement for ERK activation coupled to CREB and upregulation of Arc synthesis. AB - Brain-derived neurotrophic factor (BDNF) is implicated in long-term synaptic plasticity in the adult hippocampus, but the cellular mechanisms are little understood. Here we used intrahippocampal microinfusion of BDNF to trigger long term potentiation (BDNF-LTP) at medial perforant path--granule cell synapses in vivo. BDNF infusion led to rapid phosphorylation of the mitogen-activated protein (MAP) kinases ERK (extracellular signal-regulated protein kinase) and p38 but not JNK (c-Jun N-terminal protein kinase). These effects were restricted to the infused dentate gyrus; no changes were observed in microdissected CA3 and CA1 regions. Local infusion of MEK (MAP kinase kinase) inhibitors (PD98059 and U0126) during BDNF delivery abolished BDNF-LTP and the associated ERK activation. Application of MEK inhibitor during established BDNF-LTP had no effect. Activation of MEK-ERK is therefore required for the induction, but not the maintenance, of BDNF-LTP. BDNF-LTP was further coupled to ERK-dependent phosphorylation of the transcription factor cAMP response element-binding protein. Finally, we investigated the expression of two immediate early genes, activity-regulated cytoskeleton-associated protein (Arc) and Zif268, both of which are required for generation of late, mRNA synthesis-dependent LTP. BDNF infusion resulted in selective upregulation of mRNA and protein for Arc. In situ hybridization showed that Arc transcripts are rapidly and extensively delivered to granule cell dendrites. U0126 blocked Arc upregulation in parallel with BDNF LTP. The results support a model in which BDNF triggers long-lasting synaptic strengthening through MEK-ERK and selective induction of the dendritic mRNA species Arc. PMID- 11880484 TI - Enhanced neurosteroid potentiation of ternary GABA(A) receptors containing the delta subunit. AB - Attenuated behavioral sensitivity to neurosteroids has been reported for mice deficient in the GABA(A) receptor delta subunit. We therefore investigated potential subunit-specific neurosteroid pharmacology of the following GABA(A) receptor isoforms in a transient expression system: alpha1beta3gamma2L, alpha1beta3delta, alpha6beta3gamma2L, and alpha6beta3delta. Potentiation of submaximal GABA(A) receptor currents by the neurosteroid tetrahydrodeoxycorticosterone (THDOC) was greatest for the alpha1beta3delta isoform. Whole-cell GABA concentration--response curves performed with and without low concentrations (30 nm) of THDOC revealed enhanced peak GABA(A) receptor currents for isoforms tested without affecting the GABA EC50. Alpha1beta3delta currents were enhanced the most (>150%), whereas the other isoform currents were enhanced 15-50%. At a higher concentration (1 microm), THDOC decreased peak alpha1beta3gamma2L receptor current amplitude evoked by GABA (1 mm) concentration jumps and prolonged deactivation but had little effect on the rate or extent of apparent desensitization. Thus the polarity of THDOC modulation depended on GABA concentration for alpha1beta3gamma2L GABA(A) receptors. However, the same protocol applied to alpha1beta3delta receptors resulted in peak current enhancement by THDOC of >800% and prolonged deactivation. Interestingly, THDOC induced pronounced desensitization in the minimally desensitizing alpha1beta3delta receptors. Single channel recordings obtained from alpha1beta3delta receptors indicated that THDOC increased the channel opening duration, including the introduction of an additional longer duration open state. Our results suggest that the GABA(A) receptor delta subunit confers increased sensitivity to neurosteroid modulation and that the intrinsic gating and desensitization kinetics of alpha1beta3delta GABA(A) receptors are altered by THDOC. PMID- 11880486 TI - Cytokines regulate microglial adhesion to laminin and astrocyte extracellular matrix via protein kinase C-dependent activation of the alpha6beta1 integrin. AB - Microglia are highly plastic cells that participate in inflammatory and injury responses within the CNS and that can migrate extensively after activation. Because astrocytes and their extracellular matrix (ECM) form a large part of the CNS parenchyma, we undertook to study the adhesive interactions between microglia and these substrates in vitro. In contrast to oligodendrocyte precursor cells, microglia formed only weak interactions with astrocytes and their ECM. On specific ECM substrates the microglia adhered strongly to fibronectin, vitronectin, and plastic but only weakly to laminin. Microglial adhesion to laminin was increased significantly by the proinflammatory cytokines TNF, IFN alpha, and IFN-gamma but was decreased by TGF-beta1, with the TGF-beta1 effect being dominant over the other cytokines. Fluorescence-activated cell sorting (FACS) analysis and immunoprecipitation showed that microglia constitutively express the alpha6beta1 integrin, a well characterized laminin receptor, and that alpha6beta1 expression levels did not change after cytokine treatment. Function blocking studies showed that microglial adhesion to laminin is mediated entirely by the alpha6beta1 integrin, strongly suggesting that the cytokine regulation of adhesion to laminin is mediated by changes in the activation state of alpha6beta1. Analysis of signaling pathways revealed that activation of alpha6beta1 is mediated by a PKC-dependent mechanism. In light of the evidence that laminin expression is upregulated after CNS injury, the findings suggest that cytokine regulation of microglial adhesion to laminin may play a fundamental role in determining the extent of microglial infiltration into and retention at the site of injury. PMID- 11880485 TI - Consequences of the stoichiometry of Slo1 alpha and auxiliary beta subunits on functional properties of large-conductance Ca2+-activated K+ channels. AB - Auxiliary beta subunits play a major role in defining the functional properties of large-conductance, Ca2+-dependent BK-type K+ channels. In particular, both the beta1 and beta2 subunits produce strong shifts in the voltage dependence of channel activation at a given Ca2+. Beta subunits are thought to coassemble with alpha subunits in a 1:1 stoichiometry, such that a full ion channel complex may contain up to four beta subunits per channel. However, previous results raise the possibility that ion channels with less than a full complement of beta subunits may also occur. The functional consequence of channels with differing stoichiometries remains unknown. Here, using expression of alpha and beta subunits in Xenopus oocytes, we show explicitly that functional BK channels can arise with less than four beta subunits. Furthermore, the results show that, for both the beta1 and beta2 subunits, each individual beta subunit produces an essentially identical, incremental effect on the voltage dependence of gating. For channels arising from alpha + beta2 subunits, the number of beta2 subunits per channel also has a substantial impact on properties of steady-state inactivation and recovery from inactivation. Thus, the stoichiometry of alpha:beta subunit assembly can play a major functional role in defining the apparent Ca2+ dependence of activation of BK channels and in influencing the availability of BK channels for activation. PMID- 11880487 TI - Alternative splicing of the beta 4 subunit has alpha1 subunit subtype-specific effects on Ca2+ channel gating. AB - Ca2+ channel beta subunits are important molecular determinants of the kinetics and voltage dependence of Ca2+ channel gating. Through direct interactions with channel-forming alpha1 subunits, beta subunits enhance expression levels, accelerate activation, and have variable effects on inactivation. Four distinct beta subunit genes each encode five homologous sequence domains (D1-5), three of which (D1, D3, and D5) undergo alternative splicing. We have isolated from human spinal cord a novel alternatively spliced beta4 subunit containing a short form of domain D1 (beta4a) that is highly homologous to N termini of Xenopus and rat beta3 subunits. The purpose of this study was to compare the gating properties of various alpha1 subunit complexes containing beta4a with those of complexes containing a beta4 subunit with a longer form of domain D1, beta4b. Expression in Xenopus oocytes revealed that, relative to alpha1A and alpha1B complexes containing beta4b, the voltage dependence of activation and inactivation of complexes containing beta4a were shifted to more depolarized potentials. Moreover, alpha1A and alpha1B complexes containing beta4a inactivated at a faster rate. Interestingly, beta4 subunit alternative splicing did not influence the gating properties of alpha1C and alpha1E subunits. Experiments with beta4 deletion mutants revealed that both the N and C termini of the beta4 subunit play critical roles in setting voltage-dependent gating parameters and that their effects are alpha1 subunit specific. Our data are best explained by a model in which distinct modes of activation and inactivation result from beta-subunit splice variant-specific interactions with an alpha1 subunit gating structure. PMID- 11880488 TI - Pituitary adenylate cyclase activating polypeptide anti-mitogenic signaling in cerebral cortical progenitors is regulated by p57Kip2-dependent CDK2 activity. AB - Generation of distinct cell types and numbers in developing cerebral cortex is subject to regulation by extracellular factors that positively or negatively control precursor proliferation. Although signals stimulating proliferation are well described, factors halting cell cycle progression are less well defined. At the molecular level, production and association of cyclins, cyclin-dependent kinases (CDKs), and CDK inhibitors (CKIs) regulate cycle progression. We now report that the endogenous peptide, pituitary adenylate cyclase activating polypeptide (PACAP), negatively regulates the cell cycle by inhibiting p57Kip2 dependent CDK2 activity in embryonic cortex. Protein levels of CDK2 and members of the CIP/KIP family of CKIs (p27Kip1, p57Kip2) were detected in developing rat cortex from embryonic day 13.5 through postnatal day 2. With advancing development, CDK2 protein levels decreased, whereas CKI expression increased, suggesting that stimulatory and inhibitory cycle proteins control cell cycle exit. Using a well defined, nonsynchronized, 8 hr precursor culture, PACAP decreased the fraction of cells crossing the G1/S boundary, inhibiting DNA synthesis by 35%. CDK2 kinase activity was inhibited 75% by PACAP, whereas kinase protein and its regulatory cyclin E subunit were unaffected. Moreover, decreased kinase activity was accompanied by a twofold increase in levels of p57Kip2 protein, but not p21Cip1 or p27Kip1, suggesting that p57Kip2 mediates PACAP anti mitogenic effects. Indeed, immunoprecipitation of CDK2 complex revealed increased p57Kip2 association with the kinase and concomitant reduction in free inhibitor after PACAP exposure, suggesting that p57Kip2 interactions directly regulate CDK2 activity. These observations establish a mechanism whereby anti-mitogenic signals actively induce cell cycle withdrawal in developing cortex. PMID- 11880490 TI - In vivo disruption of Xenopus CLOCK in the retinal photoreceptor cells abolishes circadian melatonin rhythmicity without affecting its production levels. AB - Xenopus laevis retinas, like retinas from all vertebrate classes, have endogenous circadian clocks that control many aspects of normal retinal physiology occurring in cells throughout all layers of the retina. The localization of the clock(s) that controls these various rhythms remains unclear. One of the best studied rhythmic events is the nocturnal release of melatonin. Photoreceptor layers can synthesize rhythmic melatonin when these cells are in isolation. However, within the intact retina, melatonin is controlled in a complex way, indicating that signals from many parts of the retina may contribute to the production of melatonin rhythmicity. To test this hypothesis, we generated transgenic tadpoles that express different levels of a dominant negative Xenopus CLOCK specifically in the retinal photoreceptors. Eyes from these tadpoles continued to produce melatonin at normal levels, but with greatly disrupted rhythmicity, the severity of which correlated with the transgene expression level. These results demonstrate that although many things contribute to melatonin production in vivo, the circadian clock localized in the retinal photoreceptors is necessary for its rhythmicity. Furthermore, these data show that the control of the level of melatonin synthesis is separable from the control of its rhythmicity and may be controlled by different molecular machinery. This type of specific "molecular lesion" allows perturbation of the clock in intact tissues and is valuable for dissection of clock control of tissue-level processes in this and other complex systems. PMID- 11880489 TI - Therapeutic effects of coenzyme Q10 and remacemide in transgenic mouse models of Huntington's disease. AB - There is substantial evidence that bioenergetic defects and excitotoxicity may play a role in the pathogenesis of Huntington's disease (HD). Potential therapeutic strategies for neurodegenerative diseases in which there is reduced energy metabolism and NMDA-mediated excitotoxicity are the administration of the mitochondrial cofactor coenzyme Q10 and the NMDA antagonist remacemide. We found that oral administration of either coenzyme Q10 or remacemide significantly extended survival and delayed the development of motor deficits, weight loss, cerebral atrophy, and neuronal intranuclear inclusions in the R6/2 transgenic mouse model of HD. The combined treatment, using coenzyme Q10 and remacemide together, was more efficacious than either compound alone, resulting in an approximately 32 and 17% increase in survival in the R6/2 and N171-82Q mice, respectively. Magnetic resonance imaging showed that combined treatment significantly attenuated ventricular enlargement in vivo. These studies further implicate defective energy metabolism and excitotoxicity in the R6/2 and N171-82Q transgenic mouse models of HD and are of interest in comparison with the outcome of a recent clinical trial examining coenzyme Q10 and remacemide in HD patients. PMID- 11880491 TI - Fast vesicle recycling supports neurotransmission during sustained stimulation at hippocampal synapses. AB - High-frequency induced short-term synaptic depression is a common feature of central synapses in which synaptic responses rapidly decrease to a sustained level. A limitation in the availability of release-ready vesicles is thought to be a major factor underlying this phenomenon. Here, we studied the kinetics of vesicle reavailability and reuse during synaptic depression at hippocampal synapses. High-intensity stimulation of neurotransmitter release was induced by hyperosmolarity, high potassium, or action potential firing at 30 Hz to produce synaptic depression. Under these conditions, synaptic transmission rapidly depressed to a plateau level that was typically 10-40% of the initial response and persisted at this level for at least 5 min regardless of the developmental stage of synapses. This nondeclining phase of transmission was partly sustained by fast recycling and reuse of synaptic vesicles even after minutes of stimulation. Simultaneous electrical recording of postsynaptic responses and styryl dye destaining showed that after an initial round of exocytosis, vesicles were available for reuse with a delay between 1 and 3 sec during 30 Hz action potential or hypertonicity-induced stimulation. During these stimulation paradigms, there was a limited mobilization of vesicles from the reserve pool. During 10 Hz stimulation, however, the extent of vesicle reuse was minimal during the first 20 sec. These results suggest a role for fast vesicle recycling as a functional homeostatic mechanism that prevents vesicle depletion and maintains synaptic responses in the face of intense stimulation. PMID- 11880492 TI - Physiological role of calcium-activated potassium currents in the rat lateral amygdala. AB - Principal neurons in the lateral nucleus of the amygdala (LA) exhibit a continuum of firing properties in response to prolonged current injections ranging from those that accommodate fully to those that fire repetitively. In most cells, trains of action potentials are followed by a slow afterhyperpolarization (AHP) lasting several seconds. Reducing calcium influx either by lowering concentrations of extracellular calcium or by applying nickel abolished the AHP, confirming it is mediated by calcium influx. Blockade of large conductance calcium-activated potassium channel (BK) channels with paxilline, iberiotoxin, or TEA revealed that BK channels are involved in action potential repolarization but only make a small contribution to the fast AHP that follows action potentials. The fast AHP was, however, markedly reduced by low concentrations of 4 aminopyridine and alpha-dendrotoxin, indicating the involvement of voltage-gated potassium channels in the fast AHP. The medium AHP was blocked by apamin and UCL1848, indicating it was mediated by small conductance calcium-activated potassium channel (SK) channels. Blockade of these channels had no effect on instantaneous firing. However, enhancement of the SK-mediated current by 1-ethyl 2-benzimidazolinone or paxilline increased the early interspike interval, showing that under physiological conditions activation of SK channels is insufficient to control firing frequency. The slow AHP, mediated by non-SK BK channels, was apamin-insensitive but was modulated by carbachol and noradrenaline. Tetanic stimulation of cholinergic afferents to the LA depressed the slow AHP and led to an increase in firing. These results show that BK, SK, and non-BK SK-mediated calcium-activated potassium currents are present in principal LA neurons and play distinct physiological roles. PMID- 11880493 TI - Evidence for a centrally located gate in the pore of a serotonin-gated ion channel. AB - Serotonin-gated ion channels (5-HT3) are members of the ligand-gated channel family, which includes channels that are opened directly by the neurotransmitter acetylcholine, GABA, glycine, or glutamate. Although there is general agreement that the second transmembrane domain (M2) lines the pore, the position of the gate in the M2 is less certain. Here, we used substituted cysteine accessibility method (SCAM) to provide new evidence for a centrally located gate that moves during channel activation. In the closed state, three cysteine substitutions, located on the extracellular side of M2, were modified by methanethiosulfonate (MTS) reagents. In contrast, 13 cysteine substitutions were modified in the open state with MTS reagents. The pattern of inhibition (every three to four substitutions) was consistent with an alpha helical structure for the middle and cytoplasmic segments of the M2 transmembrane domain. Unexpectedly, open-state modification of two amino acids in the center of M2 with three different MTS reagents prevented channels from fully closing in the absence of neurotransmitter. Our results are consistent with a model in which the central region of the M2 transmembrane domain is inaccessible in the closed state and moves during channel activation. PMID- 11880494 TI - The mouse Crx 5'-upstream transgene sequence directs cell-specific and developmentally regulated expression in retinal photoreceptor cells. AB - Crx, an Otx-like homeobox gene, is expressed primarily in the photoreceptors of the retina and in the pinealocytes of the pineal gland. The CRX homeodomain protein is a transactivator of many photoreceptor/pineal-specific genes in vivo, such as rhodopsin and the cone opsins. Mutations in Crx are associated with the retinal diseases, cone-rod dystrophy-2, retinitis pigmentosa, and Leber's congenital amaurosis, which lead to loss of vision. We have generated transgenic mice, using 5'- and/or 3'-flanking sequences from the mouse Crx homeobox gene fused to the beta-galactosidase (lacZ) reporter gene, and we have investigated the promoter function of the cell-specific and developmentally regulated expression of Crx. All of the independent transgenic lines commonly showed lacZ expression in the photoreceptor cells of the retina and in the pinealocytes of the pineal gland. We characterized the transgenic lines in detail for cell specific lacZ expression patterns by 5-bromo-4-chloro-3-indolyl beta-D galactoside staining and lacZ immunostaining. The lacZ expression was observed in developing and developed photoreceptor cells. This observation was confirmed by coimmunostaining of dissociated retinal cells with the lacZ and opsin antibodies. The ontogeny analysis indicated that the lacZ expression completely agrees with a temporal expression pattern of Crx during retinal development. This study demonstrates that the mouse Crx 5'-upstream genomic sequence is capable of directing a cell-specific and developmentally regulated expression of Crx in photoreceptor cells. PMID- 11880495 TI - Calcium secretion coupling at calyx of Held governed by nonuniform channel vesicle topography. AB - Phasic transmitter release at synapses in the mammalian CNS is regulated by local [Ca2+] transients, which control the fusion of readily releasable vesicles docked at active zones (AZs) in the presynaptic membrane. The time course and amplitude of these [Ca2+] transients critically determine the time course and amplitude of the release and thus the frequency and amplitude tuning of the synaptic connection. As yet, the spatiotemporal nature of the [Ca2+] transients and the number and location of release-controlling Ca2+ channels relative to the vesicles, the "topography" of the release sites, have remained elusive. We used a time-dependent model to simulate Ca2+ influx, three-dimensional buffered Ca2+ diffusion, and the binding of Ca2+ to the release sensor. The parameters of the model were constrained by recent anatomical and biophysical data of the calyx of Held. Comparing the predictions of the model with previously measured release probabilities under a variety of experimental conditions, we inferred which release site topography is likely to operate at the calyx: At each AZ one or a few clusters of Ca2+ channels control the release of the vesicles. The distance of a vesicle to the cluster(s) varies across the multiple release sites of a single calyx (ranging from 30 to 300 nm; average approximately 100 nm). Assuming this topography, vesicles in different locations are exposed to different [Ca2+] transients, with peak amplitudes ranging from 0.5 to 40 microm (half-width approximately 400 microsec) during an action potential. Consequently the vesicles have different release probabilities ranging from <0.01 to 1. We demonstrate how this spatially heterogeneous release probability creates functional advantages for synaptic transmission. PMID- 11880496 TI - Phosphatidylinositol 4,5-bisphosphate modifies tubulin participation in phospholipase Cbeta1 signaling. AB - Tubulin forms the microtubule and regulates certain G-protein-mediated signaling pathways. Both functions rely on the GTP-binding properties of tubulin. Signal transduction through Galpha(q)-regulated phospholipase Cbeta1 (PLCbeta1) is activated by tubulin through a direct transfer of GTP from tubulin to Galpha(q). However, at high tubulin concentrations, inhibition of PLCbeta1 is observed. This report demonstrates that tubulin inhibits PLCbeta1 by binding the PLCbeta1 substrate phosphatidylinositol 4,5-bisphosphate (PIP2). Tubulin binding of PIP2 was specific, because PIP2 but not phosphatidylinositol 3,4,5-trisphosphate, phosphatidylinositol 3-phosphate, phosphatidylinositol, phosphatidylcholine, phosphatidylethanolamine, or inositol 1,4,5-trisphosphate inhibited microtubule assembly. PIP2 did not affect GTP binding or GTP hydrolysis by tubulin. Muscarinic agonists promoted microtubule depolymerization and translocation of tubulin to the plasma membrane. PIP2 augmented this process in both Sf9 cells, containing a recombinant PLCbeta1 pathway, and SK-N-SH neuroblastoma cells. Colocalization of tubulin and PIP2 at the plasma membrane was demonstrated with confocal laser immunofluorescence microscopy. Although tubulin bound to both Galpha(q) and PLCbeta1, PIP2 facilitated the interaction between tubulin and PLCbeta1 but not that between tubulin and Galpha(q). However, PIP2 did augment formation of tubulin--Galpha(q)-PLCbeta1 complexes. Subsequent to potentiating PLCbeta1 activation, sustained agonist-independent membrane binding of tubulin at PIP2- and PLCbeta1-rich sites appeared to inhibit Galpha(q) coupling to PLCbeta1. Furthermore, colchicine increased membrane-associated tubulin and also inhibited PLCbeta1 activity in SK-N-SH cells. Thus, tubulin, depending on local membrane concentration, may serve as a positive or negative regulator of phosphoinositide hydrolysis. Rapid changes in membrane lipid composition or in the cytoskeleton might modify neuronal signaling through such a mechanism. PMID- 11880498 TI - The cannabinoid CB1 receptor mediates retrograde signals for depolarization induced suppression of inhibition in cerebellar Purkinje cells. AB - Action potential firing or depolarization of the postsynaptic neuron can induce a transient suppression of inhibitory synaptic inputs to the depolarized neuron in the cerebellum and hippocampus. This phenomenon, termed depolarization-induced suppression of inhibition (DSI), is initiated postsynaptically by an elevation of intracellular Ca2+ concentration ([Ca2+]i) and is expressed presynaptically as a suppression of the transmitter release. It is, therefore, thought that some retrograde signal must exist from the depolarized postsynaptic neurons to the presynaptic terminals. Recent studies on hippocampal neurons have revealed that endogenous cannabinoids (endocannabinoids) play a key role as a retrograde messenger. There are, however, conflicting reports that glutamate may be a candidate retrograde messenger for cerebellar DSI that acts on presynaptic group II metabotropic glutamate receptors (mGluRs). In this study, we examined whether endocannabinoids mediate retrograde signal for cerebellar DSI. We recorded IPSCs from Purkinje cells by stimulating putative basket cell axons in mouse cerebellar slices. DSI was readily induced in evoked IPSCs by a depolarizing pulse train. We found that DSI was completely occluded by a cannabinoid agonist, WIN55,212-2, was totally eliminated by a specific antagonist of the type 1 cannabinoid (CB1) receptor, SR141716A, and was deficient in the CB1 knock-out mouse. In contrast, a group II mGluR-specific agonist, (2S,2'R,3'R)-2-(2',3' dicarboxycyclopropyl)glycine, did not completely occlude DSI, and an mGluR antagonist, (RS)-alpha-methyl-4-carboxyphenylglycine, had no depressant effect on DSI. These results clearly indicate that the CB1 receptor mediates retrograde signal for DSI in cerebellar Purkinje cells. PMID- 11880497 TI - Inhibition of intracellular cholesterol transport alters presenilin localization and amyloid precursor protein processing in neuronal cells. AB - Generation of amyloid-beta (Abeta) from the amyloid precursor protein (APP) requires proteolytic cleavage by two proteases, beta- and gamma-secretase. Several lines of evidence suggest a role for cholesterol on secretase activities, although the responsible cellular mechanisms remain unclear. Here we show that alterations in cholesterol transport from late endocytic organelles to the endoplasmic reticulum have important consequences for both APP processing and the localization of gamma-secretase-associated presenilins (PS). Exposure of neuronal cells to cholesterol transport-inhibiting agents resulted in a marked decrease in beta-cleavage of full-length APP. In contrast, gamma-secretase activity on APP C terminal fragments was enhanced, increasing the production of both Abeta40 and Abeta42. Remarkably, retention of cholesterol in endosomal/lysosomal compartments induced PS1 and PS2 to accumulate in Rab7-positive vesicular organelles implicated in cholesterol sorting. Accumulation of PS in vesicular compartments was prominent in both Chinese hamster ovary cells deficient in Niemann-Pick C1 protein as well as in neuronal cells exposed to the cholesterol transport inhibiting agent U18666A. Because Abeta42 also localized to PS1-containing vesicular compartments, organelles involved in cholesterol transport might represent an important site for gamma-secretase activity. Our results suggest that the subcellular distribution of cholesterol may be an important factor in how cholesterol alters Abeta production and the risk of Alzheimer's disease. PMID- 11880499 TI - In vivo properties of the Drosophila inebriated-encoded neurotransmitter transporter. AB - Altering neurotransmitter levels within the nervous system can cause profound changes in behavior and neuronal function. Neurotransmitter transporters play important roles in regulating neurotransmitter levels by performing neurotransmitter reuptake. It was previously shown that mutations in the Drosophila inebriated (ine)-encoded neurotransmitter transporter cause increased neuronal excitability. Here we report a further functional characterization of Ine. First we show that Ine functions in the short-term (time scale of minutes to a few hours) to regulate neuronal excitability. Second, we show that Ine is able to control excitability from either neurons or glia cells. Third, we show that overexpression of Ine reduces neuronal excitability. Overexpression phenotypes of ine include: delayed onset of long-term facilitation and increased failure rate of transmitter release at the larval neuromuscular junction, reduced amplitude of larval nerve compound action potentials, suppression of the leg-shaking behavior of mutants defective in the Shaker-encoded potassium channel, and temperature sensitive paralysis. Each of these overexpression phenotypes closely resembles those of loss of function mutants in the para-encoded sodium channel. These data raise the possibility that Ine negatively regulates neuronal sodium channels, and thus that the substrate neurotransmitter of Ine positively regulates sodium channels. PMID- 11880500 TI - Characterization of central inhibitory muscarinic autoreceptors by the use of muscarinic acetylcholine receptor knock-out mice. AB - Forebrain muscarinic acetylcholine (ACh) receptors (mAChRs; M1-M5) are predicted to play important roles in many fundamental central functions, including higher cognitive processes and modulation of extrapyramidal motor activity. Synaptic ACh levels are known to be regulated by the activity of presynaptic muscarinic autoreceptors mediating inhibition of ACh release. Primarily because of the use of ligands with limited receptor subtype selectivity, classical pharmacological studies have led to conflicting results regarding the identity of the mAChR subtypes mediating this activity in different areas of the brain. To investigate the molecular identity of hippocampal, cortical, and striatal inhibitory muscarinic autoreceptors in a more direct manner, we used genetically altered mice lacking functional M2 and/or M4 mAChRs [knock-out (KO) mice]. After labeling of cellular ACh pools with [3H]choline, potassium-stimulated [3H]ACh release was measured in superfused brain slices, either in the absence or the presence of muscarinic drugs. The nonsubtype-selective muscarinic agonist, oxotremorine (0.1 10 microm), inhibited potassium-stimulated [3H]ACh release in hippocampal, cortical, and striatal slices prepared from wild-type mice by up to 80%. This activity was totally abolished in tissues prepared from M2-M4 receptor double KO mice. Strikingly, release studies with brain slices from M2 and M4 receptor single KO mice indicated that autoinhibition of ACh release is mediated primarily by the M2 receptor in hippocampus and cerebral cortex, but predominantly by the M4 receptor in the striatum. These results, together with additional receptor localization studies, support the novel concept that autoinhibition of ACh release involves different mAChRs in different regions of the brain. PMID- 11880501 TI - Downregulation of otospiralin, a novel inner ear protein, causes hair cell degeneration and deafness. AB - Mesenchymal nonsensory regions of the inner ear are important structures surrounding the neurosensory epithelium that are believed to participate in the ionic homeostasis of the cochlea and vestibule. We report here the discovery of otospiralin, an inner ear-specific protein that is produced by fibrocytes from these regions, including the spiral ligament and spiral limbus in the cochlea and the maculae and semicircular canals in the vestibule. Otospiralin is a novel 6.4 kDa protein of unknown function that shares a protein motif with the gag p30 core shell nucleocapsid protein of type C retroviruses. To evaluate its functional importance, we downregulated otospiralin by cochlear perfusion of antisense oligonucleotides in guinea pigs. This led to a rapid threshold elevation of the compound action potentials and irreversible deafness. Cochlear examination by transmission electron microscopy revealed hair cell loss and degeneration of the organ of Corti. This demonstrates that otospiralin is essential for the survival of the neurosensory epithelium. PMID- 11880502 TI - Genetic dysmyelination alters the molecular architecture of the nodal region. AB - We have examined the molecular organization of axons in the spinal cords of myelin-deficient (md) rats, which have profound CNS dysmyelination associated with oligodendrocyte cell death. Although myelin sheaths are rare, most large axons are at least partially surrounded by oligodendrocyte processes. At postnatal day 7 (P7), almost all node-like clusters of voltage-gated Na+ channels and ankyrinG are adjacent to axonal segments ensheathed by oligodendrocytes, but at P21, many node-like clusters are found in axonal segments that lack oligodendrocyte ensheathment. In P21 wild-type (WT) rats, the voltage-gated Na+ channels Na(v)1.2, Na(v)1.6, and Na(v)1.8, are found in different subpopulations of myelinated axons, and md rats have a similar distribution. The known molecular components of paranodes--contactin, Caspr, and neurofascin 155--are not clustered in md spinal cords, and no septate-like junctions between oligodendrocyte processes and axons are found by electron microscopy. Furthermore, Kv1.1 and Kv1.2 K+ channels are not spatially segregated from the node-like clusters of Na+ channels in md rats, in contrast to their WT littermates. These results suggest the following: node-like clusters of voltage-gated Na+ channels and ankyrinG form adjacent to ensheathed axonal segments even in the absence of a myelin sheath; these clusters persist after oligodendrocyte cell death; dysmyelination does not alter the expression of different nodal of voltage-gated Na+ channels; the absence of paranodes results in the mislocalization of neurofascin155, contactin, and Caspr, and the aberrant localization of Kv1.1 and Kv1.2. PMID- 11880503 TI - Caspase-3-dependent proteolytic cleavage of protein kinase Cdelta is essential for oxidative stress-mediated dopaminergic cell death after exposure to methylcyclopentadienyl manganese tricarbonyl. AB - In the present study, we characterized oxidative stress-dependent cellular events in dopaminergic cells after exposure to an organic form of manganese compound, methylcyclopentadienyl manganese tricarbonyl (MMT). In pheochromocytoma cells, MMT exposure resulted in rapid increase in generation of reactive oxygen species (ROS) within 5--15 min, followed by release of mitochondrial cytochrome C into cytoplasm and subsequent activation of cysteine proteases, caspase-9 (twofold to threefold) and caspase-3 (15- to 25-fold), but not caspase-8, in a time- and dose dependent manner. Interestingly, we also found that MMT exposure induces a time- and dose-dependent proteolytic cleavage of native protein kinase Cdelta (PKCdelta, 72-74 kDa) to yield 41 kDa catalytically active and 38 kDa regulatory fragments. Pretreatment with caspase inhibitors (Z-DEVD-FMK or Z-VAD-FMK) blocked MMT-induced proteolytic cleavage of PKCdelta, indicating that cleavage is mediated by caspase-3. Furthermore, inhibition of PKCdelta activity with a specific inhibitor, rottlerin, significantly inhibited caspase-3 activation in a dose-dependent manner along with a reduction in PKCdelta cleavage products, indicating a possible positive feedback activation of caspase-3 activity by PKCdelta. The presence of such a positive feedback loop was also confirmed by delivering the catalytically active PKCdelta fragment. Attenuation of ROS generation, caspase-3 activation, and PKCdelta activity before MMT treatment almost completely suppressed DNA fragmentation. Additionally, overexpression of catalytically inactive PKCdelta(K376R) (dominant-negative mutant) prevented MMT induced apoptosis in immortalized mesencephalic dopaminergic cells. For the first time, these data demonstrate that caspase-3-dependent proteolytic activation of PKCdelta plays a key role in oxidative stress-mediated apoptosis in dopaminergic cells after exposure to an environmental neurotoxic agent. PMID- 11880504 TI - Folic acid deficiency and homocysteine impair DNA repair in hippocampal neurons and sensitize them to amyloid toxicity in experimental models of Alzheimer's disease. AB - Recent epidemiological and clinical data suggest that persons with low folic acid levels and elevated homocysteine levels are at increased risk of Alzheimer's disease (AD), but the underlying mechanism is unknown. We tested the hypothesis that impaired one-carbon metabolism resulting from folic acid deficiency and high homocysteine levels promotes accumulation of DNA damage and sensitizes neurons to amyloid beta-peptide (Abeta) toxicity. Incubation of hippocampal cultures in folic acid-deficient medium or in the presence of methotrexate (an inhibitor of folic acid metabolism) or homocysteine induced cell death and rendered neurons vulnerable to death induced by Abeta. Methyl donor deficiency caused uracil misincorporation and DNA damage and greatly potentiated Abeta toxicity as the result of reduced repair of Abeta-induced oxidative modification of DNA bases. When maintained on a folic acid-deficient diet, amyloid precursor protein (APP) mutant transgenic mice, but not wild-type mice, exhibited increased cellular DNA damage and hippocampal neurodegeneration. Levels of Abeta were unchanged in the brains of folate-deficient APP mutant mice. Our data suggest that folic acid deficiency and homocysteine impair DNA repair in neurons, which sensitizes them to oxidative damage induced by Abeta. PMID- 11880505 TI - Blockade of microglial activation is neuroprotective in the 1-methyl-4-phenyl 1,2,3,6-tetrahydropyridine mouse model of Parkinson disease. AB - 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) damages the nigrostriatal dopaminergic pathway as seen in Parkinson's disease (PD), a common neurodegenerative disorder with no effective protective treatment. Consistent with a role of glial cells in PD neurodegeneration, here we show that minocycline, an approved tetracycline derivative that inhibits microglial activation independently of its antimicrobial properties, mitigates both the demise of nigrostriatal dopaminergic neurons and the formation of nitrotyrosine produced by MPTP. In addition, we show that minocycline not only prevents MPTP induced activation of microglia but also the formation of mature interleukin 1beta and the activation of NADPH-oxidase and inducible nitric oxide synthase (iNOS), three key microglial-derived cytotoxic mediators. Previously, we demonstrated that ablation of iNOS attenuates MPTP-induced neurotoxicity. Now, we demonstrate that iNOS is not the only microglial-related culprit implicated in MPTP-induced toxicity because mutant iNOS-deficient mice treated with minocycline are more resistant to this neurotoxin than iNOS-deficient mice not treated with minocycline. This study demonstrates that microglial-related inflammatory events play a significant role in the MPTP neurotoxic process and suggests that minocycline may be a valuable neuroprotective agent for the treatment of PD. PMID- 11880506 TI - Proteolipid protein gene modulates viability and phenotype of neurons. AB - Overexpression or lack of expression of proteolipid protein (PLP) gene by oligodendrocytes causes axonal pathology. It is unclear whether dysfunction of the PLP gene mediates its effects directly on neurons or indirectly by abnormal formation of myelin sheaths. We performed experiments using cocultures and conditioned media (CM) to test the direct effect of PLP gene expression on neurons. Non-glial cell lines were stably transfected with PLP or DM20 (an alternate splice variant of PLP) cDNAs. Immunocytochemistry and enhanced green fluorescent protein expression showed that translated products were synthesized and inserted into the plasma membrane in proper conformation. The number of surviving dorsal root ganglion (DRG) neurons was significantly less than controls when cocultured for 5 d with PLP-expressing cells. The number of degenerating neurons increased in a dose-dependent manner corresponding to increasing numbers of PLP-expressing cells. However, the number of surviving DRG neurons cocultured with DM20-expressing cells was comparable to that of controls, indicating that PLP-specific products contributed to decreased neuron survival. When DRG neurons were cultured with CM from PLP- or DM20-expressing cells, significantly fewer neurons survived with CM of PLP- but not DM20-expressing cells. This suggests that secreted factors from PLP-expressing cells contribute to neuronal death. Increased neuronal death found with PLP-expressing cells cannot be attributed to density-dependent artifacts, because in each experiment the density of different cell lines was similar. This effect of CM may be mediated by a negative pH shift elicited from PLP but not DM20 expression. These results indicate that PLP gene products directly modulate neuron viability. PMID- 11880507 TI - Adult rodent neurogenic regions: the ventricular subependyma contains neural stem cells, but the dentate gyrus contains restricted progenitors. AB - Neurogenesis persists in two adult brain regions: the ventricular subependyma and the subgranular cell layer in the hippocampal dentate gyrus (DG). Previous work in many laboratories has shown explicitly that multipotential, self-renewing stem cells in the subependyma are the source of newly generated migrating neurons that traverse the rostral migratory stream and incorporate into the olfactory bulb as interneurons. These stem cells have been specifically isolated from the subependyma, and their properties of self-renewal and multipotentiality have been demonstrated in vitro. In contrast, it is a widely held assumption that the "hippocampal" stem cells that can be isolated in vitro from adult hippocampus reside in the neurogenic subgranular layer and represent the source of new granule cell neurons, but this has never been tested directly. Primary cell isolates derived from the precise microdissection of adult rodent neurogenic regions were compared using two very different commonly used culture methods: a clonal colony-forming (neurosphere) assay and a monolayer culture system. Importantly, both of these culture methods generated the same conclusion: stem cells can be isolated from hippocampus-adjacent regions of subependyma, but the adult DG proper does not contain a population of resident neural stem cells. Indeed, although the lateral ventricle and other ventricular subependymal regions directly adjacent to the hippocampus contain neural stem cells that exhibit long term self-renewal and multipotentiality, separate neuronal and glial progenitors with limited self-renewal capacity are present in the adult DG, suggesting that neuron-specific progenitors and not multipotential stem cells are the source of newly generated DG neurons throughout adulthood. PMID- 11880508 TI - Growth cone pathfinding and filopodial dynamics are mediated separately by Cdc42 activation. AB - Although evidence exists that activation of the Rho family GTPase Cdc42 affects axonal development, its specific roles within a growth cone are not well delineated. To evaluate the model that Cdc42 activation regulates growth cone navigation by promoting filopodial activity, we adopted a live analysis strategy that uses transgenic Drosophila lines in which neurons coexpressed constitutively active Cdc42 (Cdc42(V12)) and membrane-targeted green fluorescent protein. We found that growth cones that displayed pathfinding defects exhibited little change in their filopodial activity, whereas others without pathfinding defects exhibited an similar50% increase in their filopodial activity. Moreover, effector loop mutations that were added to the constitutively active Cdc42 (Cdc42(V12C40) and Cdc42(V12A37)) exerted little influence over filopodial activity caused by Cdc42 activation but suppressed the pathfinding defects of the growth cones. Together, these data suggest that Cdc42 controls filopodial activity in axonal growth cones independently of its effects on their pathfinding. PMID- 11880509 TI - Extended plasticity of visual cortex in dark-reared animals may result from prolonged expression of cpg15-like genes. AB - cpg15 is an activity-regulated gene that encodes a membrane-bound ligand that coordinately regulates growth of apposing dendritic and axonal arbors and the maturation of their synapses. These properties make it an attractive candidate for participating in plasticity of the mammalian visual system. Here we compare cpg15 expression during normal development of the rat visual system with that seen in response to dark rearing, monocular blockade of retinal action potentials, or monocular deprivation. Our results show that the onset of cpg15 expression in the visual cortex is coincident with eye opening, and it increases until the peak of the critical period at postnatal day 28 (P28). This early expression is independent of both retinal activity and visual experience. After P28, a component of cpg15 expression in the visual cortex, lateral geniculate nucleus (LGN), and superior colliculus (SC) develops a progressively stronger dependence on retinally driven action potentials. Dark rearing does not affect cpg15 mRNA expression in the LGN and SC at any age, but it does significantly affect its expression in the visual cortex from the peak of the critical period and into adulthood. In dark-reared rats, the peak level of cpg15 expression in the visual cortex at P28 is lower than in controls. Rather than showing the normal decline with maturation, these levels are maintained in dark-reared animals. We suggest that the prolonged plasticity in the visual cortex that is seen in dark-reared animals may result from failure to downregulate genes such as cpg15 that could promote structural remodeling and synaptic maturation. PMID- 11880510 TI - Retrograde repression of growth-associated protein-43 mRNA expression in rat cortical neurons. AB - Corticospinal neurons support rapid growth of axons toward spinal cord targets in the perinatal period. Initial axon growth is accompanied by elevated expression of growth-associated protein-43 (GAP-43), which then declines in postnatal development. To investigate whether expression of GAP-43 mRNA is regulated by retrograde signals, we injected colchicine into the corticospinal tract to block retrograde axonal transport during a time when GAP-43 is normally declining in corticospinal neurons. Colchicine caused a prolongation of high GAP-43 mRNA expression in neurons located in layer V (but not other layers) of sensorimotor cortex. We next used osmotic minipumps to infuse soluble adult spinal cord extract into the sensorimotor cortex. This resulted in a premature downregulation of GAP-43 mRNA in identified corticospinal neurons. GAP-43 repressive activity was found in extracts of the spinal cord tissue as young as postnatal day 8. The effect of spinal cord extract in vivo was not mimicked by adult cerebellar or muscle extracts. Cultures of postnatal cortical neurons also underwent downregulation of GAP-43 mRNA when treated with spinal cord extract. Activation of cAMP signaling also repressed GAP-43 mRNA in cortical cultures, and the repressive effect of spinal cord extract was diminished by an adenyl cyclase inhibitor. Thus, GAP-43 mRNA may be downregulated late in development by a target derived retrograde repressive factor, and this effect may be mediated by cAMP second messenger signaling. PMID- 11880511 TI - Long-term depression in the developing hippocampus: low induction threshold and synapse nonspecificity. AB - It was observed that the use of paired-pulse afferent stimulation as test stimulation (0.1-0.02 Hz) in the hippocampal CA1 area in young (1-2 week) rats, but not in older ones, led to declining synaptic activity. We show that such very low-frequency stimulation leads to long-term depression (LTD) initiated by activation of NMDA receptor channels and/or T-type voltage-dependent calcium channels. The depression is initiated within three or four such stimuli, and <10 are sufficient to induce a notable long-term effect. When the paired-pulse stimulation exceeded threshold for postsynaptic spike activation, the depression was preceded by an NMDA receptor-dependent potentiation. Irrespective of whether homosynaptic potentiation or depression occurred, the paired pulse stimulation also induced depression in neighboring synapses alternately activated by single stimuli. These results point to a very high sensitivity for induction of synaptic depression during the neonatal period. They also support the notion that a brief rise in postsynaptic calcium can induce long-term potentiation (LTP) or LTD, a larger rise more likely to induce LTP, as well as that a prolonged modest increase produces selectively only LTD. PMID- 11880512 TI - Synapse formation in the absence of cell bodies requires protein synthesis. AB - Protein synthesis at distal synaptic sites is thought to play a critical role in long-term synaptic plasticity at preexisting connections. We tested whether protein synthesis in distal neuritic processes contributes to the formation of new synaptic connections by Aplysia neurons regenerating in cell culture after removing their cell bodies. Removal of either the sensory neuron (SN) or motor cell L7 cell body did not affect the formation of synaptic connections during the next 48--72 hr period. Increases in synaptic efficacy after removal of the SN cell body was accompanied by neurite growth and an increase in the number of SN varicosities contacting L7. The increases in synaptic efficacy and the number of SN varicosities were blocked by anisomycin, a protein synthesis inhibitor. The initial formation of synaptic connections was not affected by the absence of the L7 cell body. In the absence of cell bodies from both presynaptic and postsynaptic cells, synaptic efficacy increased for 48 hr and was blocked reversibly by anisomycin. These results support the idea that distal neuritic processes contain stable mRNAs and the macromolecular machinery for protein synthesis that are required for the formation of new synaptic connections. PMID- 11880513 TI - Cloning and characterization of glioma BK, a novel BK channel isoform highly expressed in human glioma cells. AB - Voltage-dependent large-conductance Ca2+-activated K+ channels (BK channels) are widely expressed in excitable and nonexcitable cells. BK channels exhibit diverse electrophysiological properties, which are attributable in part to alternative splicing of their alpha-subunits. BK currents have been implicated in the growth control of glial cells, and BK channels with novel biophysical properties have recently been characterized in human glioma cells. Here we report the isolation, cloning, and functional characterization of glioma BK (gBK), a novel splice isoform of hSlo, the gene that encodes the alpha-subunits of human BK channels. The primary sequence of gBK is 97% identical to its closest homolog hbr5, but it contains an additional 34-amino-acid exon at splice site 2 in the C-terminal tail of BK channels. hSlo transcripts containing this novel exon are expressed ubiquitously in various normal tissues as well as in neoplasmic samples, suggesting that the novel exon may modulate important physiological functions of BK channels. Expression of gBK in Xenopus oocytes gives rise to iberiotoxin sensitive (IbTX) currents, with an IC(50) for IbTX of 5.7 nm and a Hill coefficient of 0.76. Single gBK channels have a unitary conductance of similar250 pS, and the currents show significantly slower activation and higher Ca2+ sensitivity than hbr5. Ca2+ sensitivity was enhanced specifically at physiologically relevant [Ca2+]i (100-500 nm). Examination of biopsies from patients with malignant gliomas has revealed specific overexpression of BK channels in gliomas compared with nonmalignant human cortical tissues. Importantly, tumor malignancy grades have correlated positively with BK channel expression, suggesting an important role for the gBK channel in glioma biology. PMID- 11880514 TI - Taste-specific neuronal ensembles in the gustatory cortex of awake rats. AB - In gustatory cortex, single-neuron activity reflects the multimodal processing of taste stimuli. Little is known, however, about the interactions between gustatory cortical (GC) neurons during tastant processing. Here, these interactions were characterized. It was found that 36% (85 of 237) of neuron pairs, including many (61%) in which one or both single units were not taste specific, produced significant cross-correlations (CCs) to a subset of tastants across a hundreds of milliseconds timescale. Significant CCs arose from the coupling between the firing rates of neurons as those rates changed through time. Such coupling significantly increased the amount of tastant-specific information contained in ensembles. These data suggest that taste-specific GC assemblies may transiently form and coevolve on a behaviorally appropriate timescale, contributing to rats' ability to discriminate tastants. PMID- 11880515 TI - The relationship between Abeta and memory in the Tg2576 mouse model of Alzheimer's disease. AB - Transgenic mice expressing mutant amyloid precursor proteins (APPs) have provided important new information about the pathogenesis of Alzheimer's disease (AD) histopathology. However, the molecular basis of memory loss in these mice is poorly understood. One of the major impediments has been the difficulty of distinguishing between age-dependent and age-independent behavioral changes. To address this issue we studied in parallel two lines of APP transgenic mice expressing comparable levels of mutant and wild-type human APP. This enabled us to identify age-independent behavioral deficits that were not specifically related to mutant APP expression. When mice with age-independent deficits were eliminated, we detected memory loss in transgenic mice expressing mutant APP (Tg2576 mice) starting at approximately 6 months, which coincided with the appearance of detergent-insoluble Abeta aggregates (Abeta(insol)). Genetically accelerating the formation of Abeta(insol) resulted in an earlier onset of memory decline. A facile interpretation of these results, namely that memory loss and Abeta(insol) were closely connected, was rejected when we extended our analysis to include older mice. No obvious correspondence between memory and Abeta(insol) was apparent in a combined group of old and young mice unless the mice were stratified by age, whereupon inverse correlations between memory and Abeta(insol) became evident. These results suggested that Abeta(insol) is a surrogate marker for small assemblies of Abeta that disrupt cognition and occur as intermediates during Abeta(insol) formation, and they are the first descriptive in vivo data supporting their role in impairing memory. These studies also provide a methodological framework within which to investigate these Abeta assemblies in vivo. PMID- 11880516 TI - Signaling mechanisms of metabotropic glutamate receptor 5 subtype and its endogenous role in a locomotor network. AB - Metabotropic glutamate receptors (mGluRs) act as modulators in the CNS of vertebrates, but their role in motor pattern generation in particular is primarily unknown. The intracellular signaling mechanisms of the group I mGluRs (mGluR1 and mGluR5), and their endogenous role in regulating locomotor pattern generation have been investigated in the spinal cord of the lamprey. Application of the group I mGluR agonist (R,S)-3,5-dihydroxyphenylglycine (DHPG) produced oscillations of the intracellular Ca2+ concentration ([Ca2+]i) in neurons. The oscillations were blocked by the mGluR5 antagonist 2-methyl-6 (phenylethynyl)pyridine (MPEP) but not by the mGluR1 antagonist 7 (hydroxyimino)cyclopropa[b]chromen-1a-carboxylate ethyl ester. These [Ca2+]i oscillations were abolished by a phospholipase C blocker and after depletion of internal Ca2+ stores by thapsigargin but did not involve protein kinase C activation. Furthermore, they were dependent on Ca2+ influx, because no [Ca2+]i oscillations were produced by DHPG in a Ca2+-free solution or after blockade of L type Ca2+ channels. The mGluR5 is activated by an endogenous release of glutamate during locomotion, and a receptor blockade by MPEP caused an increase in the burst frequency. Thus, our results show that mGluR5 induces [Ca2+]i oscillations and regulates the activity of locomotor networks through endogenous activation. PMID- 11880517 TI - Colocalized neuropeptides activate a central pattern generator by acting on different circuit targets. AB - In the presence of descending modulatory inputs, the stomatogastric ganglion (STG) of the lobster Homarus americanus generates a triphasic motor pattern, the pyloric rhythm. Red pigment-concentrating hormone (RPCH) and Cancer borealis tachykinin-related peptide (CabTRP) are colocalized in a pair of fibers that project into the neuropil of the STG. When the STG was isolated from anterior ganglia modulatory inputs, the lateral pyloric (LP) and pyloric (PY) neurons became silent, whereas the anterior burster (AB) and pyloric dilator (PD) neurons were rhythmically active at a low frequency. Exogenous application of 10(-6) m RPCH activated the LP neuron but not the PY neurons; 10(-6) m CabTRP activated the PY neurons but not the LP neuron. The actions of RPCH on the LP neuron and CabTRP on the PY neurons persisted when the rhythmic drive from the PD and AB neurons was removed, suggesting that the LP and PY neurons are direct targets for RPCH and CabTRP respectively. Coapplication of 10(-6) m RPCH and 10(-6) m CabTRP elicited triphasic motor patterns with phase relationships resembling those in a preparation with modulatory inputs intact. In summary, cotransmitters acting on different network targets act cooperatively to activate a complete central pattern-generating circuit. PMID- 11880518 TI - Neuronal activity in substantia nigra pars reticulata during target selection. AB - Complex visual scenes require that a target for an impending saccadic eye movement be selected from a number of possible targets. We investigated whether changing the number of stimuli from which a target would be identified altered the activity of substantia nigra pars reticulata (SNr) neurons of the basal ganglia (BG) and how such changes might contribute to changes we observed previously in the superior colliculus (SC). One, two, four, or eight visual stimuli appeared on random trials while monkeys fixated a centrally located spot. After a delay, one of the stimuli in the array changed luminance, indicating that it was the saccade target. We found that SNr neurons that had a pause in tonic activity after target onset and when the saccade was made to the target showed a modulation of activity during the multitarget task. Because the number of stimuli in the array increased from one to eight, the initial pause after the onset of the visual stimulus decreased. Activity during the preselection delay was reduced but was independent of the number of possible targets present. When one of the stimuli was identified as the saccade target, but before the saccade was made, we found a sharp decline in activity. This decline was related to the monkey's selecting the target rather than the luminance change identifying the target, because on error trials, when the luminance changed but a saccade was not made to the target, the activity did not decline. The decline for the preferred target location was also accompanied by a lesser decline for adjacent locations. Our findings indicate that SNr activity changes with target selection as it does with saccade initiation and that the SNr could make substantial, direct contributions to the SC at both times. The pause in SNr activity with target selection is consistent with the hypothesis that BG provide a disinhibition for the selection of desired movements. PMID- 11880519 TI - Combinatorial and cross-fiber averaging transform muscle electrical responses with a large stochastic component into deterministic contractions. AB - Pyloric muscles of the stomatogastric neuromuscular system of the lobster Panulirus interruptus produce highly deterministic (range, less than +/- 6% of mean amplitude) contractions in response to motor nerve stimulation with unchanging spike bursts containing physiological (5-10) spike numbers. Intracellular recordings of extrajunctional potentials (EJPs) evoked in these muscles by motor nerve stimulation revealed a large, apparently stochastic amplitude variation (range, +/- 36% of mean amplitude). These observations raised the question of how do electrical responses with a large amplitude variation give rise to deterministic muscle output? We show here that this question is likely resolved by (1) combinatorial averaging within individual muscle fibers of the multiple EJPs that occur in motor neuron bursts, and (2) averaging across muscle fibers whose electrical responses are uncorrelated. Synapses with high inherent variability are also present in vertebrate CNSs. Combinatorial averaging in multispike inputs would also reduce variation in postsynaptic response at these synapses. The data reported here provide further support that bursting presynaptic activity could make such synapses functionally deterministic as well. PMID- 11880520 TI - Selective behavioral and neurochemical effects of cholinergic lesions produced by intrabasalis infusions of 192 IgG-saporin on attentional performance in a five choice serial reaction time task. AB - The effects of the cholinergic immunotoxin 192 IgG-saporin (SAP) (0.0, 0.15, or 0.45 microg/microl; 0.5 microl/hemisphere) infused into the area of the nucleus basalis magnocellularis (NBM) of rats were tested in a five-choice serial reaction time task (5CSRTT) designed to assess visual attention. The effects of this manipulation on acetylcholine efflux in the medial frontal cortex were determined using in vivo microdialysis during the 5CSRTT. Rats with extensive lesions of the NBM (SAP HIGH) showed an array of behavioral deficits in the 5CSRTT hypothesized to represent deficits in central executive function that were associated with severe deficits in accuracy. Lengthening the stimulus duration ameliorated these deficits. Rats with restricted lesions of the NBM (SAP LOW) showed impairments over time on task when tested under standard conditions that were exacerbated by increases in the event rate. The number of choline acetyltransferase-immunoreactive cells in the area of the NBM but not the vertical limb of the diagonal band correlated significantly with accuracy in the task. SAP HIGH rats had significantly lower levels of cortical acetylcholine (ACh) efflux relative to SHAM both before and during the 5CSRTT. SAP LOW rats showed significantly higher levels of cortical ACh efflux before but not during the 5CSRTT. Cortical ACh efflux increased in all rats with the onset of the attentional task. These data provide the first direct evidence for a relationship between selective damage in the basal forebrain with decreased cortical ACh efflux and impaired attentional function. PMID- 11880521 TI - Hippocampal long-term potentiation is reduced by chronic opiate treatment and can be restored by re-exposure to opiates. AB - Chronic exposure to opiates eventually leads to drug addiction, which is believed to involve maladaptive changes in brain function, but the underlying neuronal mechanisms remain primarily unknown. Given the known effects of opiates such as morphine and heroin on hippocampal function, we investigated the potential effect of chronic opiate treatment on long-term potentiation (LTP) at CA1 synapses in rat hippocampus, a leading experimental model for studying synaptic plasticity. Our results revealed that chronic exposure of rats to morphine or heroin, which induced severe drug tolerance and dependence, markedly reduced the capacity of hippocampal CA1 LTP during the period of drug withdrawal (from approximately 190% in control to approximately 120%). More interestingly, the capacity of LTP could be restored to the normal level by re-exposure of the animals to opiates, indicating that the synaptic function was already adapted to opiates. Morris water maze test, which measures behavioral consequences of synaptic plasticity, showed parallel learning deficits after chronic exposure to opiates. Moreover, the opiate-reduced LTP could also be restored by inhibitors of cAMP-dependent protein kinase A (PKA), suggesting that upregulation of cAMP pathway was likely one of the underlying mechanisms of the observed phenomena. These findings demonstrated that chronic opiate treatment can significantly modulate synaptic plasticity in the hippocampus, leading to an opiate dependence of the plasticity. PMID- 11880522 TI - Spatial working memory and the brainstem cholinergic innervation to the anterior thalamus. AB - The anteroventral thalamic nucleus (AV) has a role in spatial memory, but the influence of the prominent brainstem cholinergic projection to this region is unknown. Here, spatial memory in a 12-arm radial maze was examined after 0.15 microl bilateral AV infusions of scopolamine. In part one, rats visited six arms singly (the phase 1 arms) and, after a 10 min delay, were allowed free choice to both phase 1 arms and the remaining six baited arms (phase 2 arms). Scopolamine (10 microg) administered during the delay increased errors to both phase 1 and phase 2 arms, whereas PBS infusions increased phase 1 arm errors only. The PBS effect was the result of inserting the internal cannulas alone and not the infusion. The same dose of scopolamine (10 microg) infused before maze testing (part two: no phase 1 arms, no delay) also impaired spatial memory over and above the effects of both PBS and no-infusion, which did not differ markedly. Part two also showed that choice latency and choice strategies were unaffected by PBS and scopolamine. Cannulation and infusion procedures in both parts one and two did not produce any negative carryover effects across multiple control (no internal cannula) sessions, and a trypan blue manipulation indicated that infusions were restricted to the AV region. This study provides the first direct evidence that the brainstem cholinergic innervation to the limbic thalamus influences learning and memory, which may have important implications for human neurological conditions such as alcohol-related disorders and schizophrenia. PMID- 11880523 TI - Facilitation by endogenous tachykinins of the NMDA-evoked release of acetylcholine after acute and chronic suppression of dopaminergic transmission in the matrix of the rat striatum. AB - Using a microsuperfusion method in vitro, the effects of the NK1, NK2, and NK3 tachykinin receptor antagonists SR140333, SR48968, and SR142801, respectively, on the NMDA-evoked release of [3H]-acetylcholine were investigated after both acute and chronic suppression of dopamine transmission in striosomes and matrix of the rat striatum. NMDA (1 mm) alone or with D-serine (10 microm) in the presence of alpha-methyl-p-tyrosine (100 microm) markedly enhanced the release of [3H] acetylcholine through a dopamine-independent inhibitory process. In both conditions, as well as after chronic 6-OHDA-induced denervation of striatal dopaminergic fibers, SR140333, SR48968, or SR142801 (0.1 microm each) reduced the NMDA-evoked release of [3H]-acetylcholine in the matrix but not in striosome enriched areas. These responses were selectively abolished by coapplication with NMDA of the respective tachykinin agonists, septide, [Lys5,MeLeu9,Nle10]NKA(4 10), or senktide. Distinct mechanisms are involved in the effects of the tachykinin antagonists because the inhibitory response of SR140333 was additive with that of either SR48968 or SR142801. In addition, the SR140333-evoked response remained unchanged, whereas those of SR48968 and SR142801 were abolished in the presence of N(G)-monomethyl-l-arginine (nitric oxide synthase inhibitor). Therefore, in the matrix but not in striosomes, the acute or chronic suppression of dopamine transmission unmasked the facilitatory effects of endogenously released substance P, neurokinin A, and neurokinin B on the NMDA-evoked release of [3H]-acetylcholine. Whereas substance P and neurokinin A are colocalized in same efferent neurons, their responses involve distinct circuits because the substance P response seems to be mediated by NK1 receptors located on cholinergic interneurons, while those of neurokinin A and neurokinin B are nitric oxide dependent. PMID- 11880524 TI - Rats fail to discriminate quinine from denatonium: implications for the neural coding of bitter-tasting compounds. AB - Recent molecular findings indicate that many different G-protein-coupled taste receptors that bind with "bitter-tasting" ligands are coexpressed in single taste receptor cells in taste buds, leading to the prediction that mammals can respond behaviorally to structurally diverse "bitter" tastants but cannot discriminate among them. However, recent in situ calcium-imaging findings imply that rat taste receptor cells are more narrowly tuned to respond to bitter-tasting compounds than had been predicted from molecular findings, suggesting that these animals can discriminate among these chemicals. Using an operant conditioning paradigm, we demonstrated that rats cannot discriminate between two structurally dissimilar bitter compounds, quinine hydrochloride and denatonium benzoate, despite the fact that these tastants are thought to stimulate different taste receptor cells. These rats were nonetheless able to show concentration-dependent avoidance responses to both compounds in brief-access tests and to discriminate among other taste stimuli, including quinine versus KCl, denatonium versus KCl, and NaCl versus KCl. Importantly, the concentrations were varied in the discrimination tests to render intensity an irrelevant cue. We conclude that denatonium and quinine produce a unitary taste sensation, leaving open the likely possibility that other compounds fall into this class. Although a broader array of compounds needs to be tested, our findings lend support to the hypothesis that there is only one qualitative type of bitterness. These results also highlight the need to confirm predictions about the downstream properties of the gustatory system, or any sensory system, based on upstream molecular and biophysical events. PMID- 11880525 TI - Persistent, exocytosis-independent silencing of release sites underlies homosynaptic depression at sensory synapses in Aplysia. AB - The synaptic connections of Aplysia sensory neurons (SNs) undergo dramatic homosynaptic depression (HSD) with only a few low-frequency stimuli. Strong and weak SN synapses, although differing in their probabilities of release, undergo HSD at the same rate; this suggests that the major mechanism underlying HSD in these SNs may not be depletion of the releasable pool of vesicles. In computational models, we evaluated alternative mechanisms of HSD, including vesicle depletion, to determine which mechanisms enable strong and weak synapses to depress with identical time courses. Of five mechanisms tested, only release independent, stimulus-dependent switching off of release sites resulted in HSD that was independent of initial synaptic strength. This conclusion that HSD is a release-independent phenomenon was supported by empirical results: an increase in Ca2+ influx caused by spike broadening with a K+ channel blocker did not alter HSD. Once induced, HSD persisted during 40 min of rest with no detectable recovery; thus, release does not recover automatically with rest, contrary to what would be expected if HSD represented an exhaustion of the exocytosis mechanism. The hypothesis that short-term HSD involves primarily a stepwise silencing of release sites, rather than vesicle depletion, is consistent with our earlier observation that HSD is accompanied by only a modest decrease in release probability, as indicated by little change in the paired-pulse ratio. In contrast, we found that there was a dramatic decrease in the paired-pulse ratio during serotonin-induced facilitation; this suggests that heterosynaptic facilitation primarily involves an increase in release probability, rather than a change in the number of functional release sites. PMID- 11880526 TI - Fast propagation of firing rates through layered networks of noisy neurons. AB - We model the propagation of neural activity through a feedforward network consisting of layers of integrate-and-fire neurons. In the presence of a noisy background current and spontaneous background firing, firing rate modulations are transmitted linearly through many layers, with a delay proportional to the synaptic time constant and with little distortion. Single neuron properties and firing statistics are in agreement with physiological data. The proposed mode of propagation allows for fast computation with population coding based on firing rates, as is demonstrated with a local motion detector. PMID- 11880527 TI - Blockade of striatal adenosine A2A receptor reduces, through a presynaptic mechanism, quinolinic acid-induced excitotoxicity: possible relevance to neuroprotective interventions in neurodegenerative diseases of the striatum. AB - The aim of the present study was to evaluate whether, and by means of which mechanisms, the adenosine A2A receptor antagonist SCH 58261 [5-amino-7-(2 phenylethyl)-2-(2-furyl)-pyrazolo[4,3-e]-1,2,4-triazolo[1,5-c]pyrimidine] exerted neuroprotective effects in a rat model of Huntington's disease. In a first set of experiments, SCH 58261 (0.01 and 1 mg/kg) was administered intraperitoneally to Wistar rats 20 min before the bilateral striatal injection of quinolinic acid (QA) (300 nmol/1 microl). SCH 58261 (0.01 but not 1 mg/kg, i.p.) did reduce significantly the effects of QA on motor activity, electroencephalographic changes, and striatal gliosis. Because QA acts by both increasing glutamate outflow and directly stimulating NMDA receptors, a second set of experiments was performed to evaluate whether SCH 58261 acted by preventing the presynaptic and/or the postsynaptic effects of QA. In microdialysis experiments in naive rats, striatal perfusion with QA (5 mm) enhanced glutamate levels by approximately 500%. Such an effect of QA was completely antagonized by pretreatment with SCH 58261 (0.01 but not 1 mg/kg, i.p.). In primary striatal cultures, bath application of QA (900 microm) significantly increased intracellular calcium levels, an effect prevented by the NMDA receptor antagonist MK-801 [(+)-5-methyl-10,11-dihydro-5H-dibenzo [a,d] cyclohepten-5,10-imine maleate]. In this model, bath application of SCH 58261 (15-200 nm) tended to potentiate QA-induced calcium increase. We conclude the following: (1) the adenosine A2A receptor antagonist SCH 58261 has neuroprotective effects, although only at low doses, in an excitotoxic rat model of HD, and (2) the inhibition of QA-evoked glutamate outflow seems to be the major mechanism underlying the neuroprotective effects of SCH 58261. PMID- 11880528 TI - Dynamics of spatial frequency tuning in macaque V1. AB - Spatial frequency tuning in the lateral geniculate nucleus of the thalamus (LGN) and primary visual cortex (V1) differ substantially. LGN responses are largely low-pass in spatial frequency, whereas the majority of V1 neurons have bandpass characteristics. To study this transformation in spatial selectivity, we measured the dynamics of spatial frequency tuning using a reverse correlation technique. We find that a large proportion of V1 cells show inseparable responses in spatial frequency and time. In several cases, tuning becomes more selective over the course of the response, and the preferred spatial frequency shifts from low to higher frequencies. Many responses also show suppression at low spatial frequencies, which correlates with the increases in response selectivity and the shifts of preferred spatial frequency. These results indicate that suppression plays an important role in the generation of bandpass selectivity in V1. PMID- 11880529 TI - Highly dissimilar behaviors mediated by a multifunctional network in the marine mollusk Tritonia diomedea. AB - Several motor networks have now been found to be multifunctional, in which one group of neurons participates in the generation of multiple behavioral motor programs. Not surprisingly, the behaviors involved are frequently closely related, often using the same or similar muscle groups. Here we describe an interneuronal network in the marine mollusk Tritonia diomedea that is involved in producing two highly dissimilar behaviors, rhythmic, muscle-based escape swimming and nonrhythmic, cilia-mediated crawling. Several observations support this conclusion. First, the dorsal swim interneurons (DSIs) of the swim central pattern generator (CPG) directly excite Pedal neuron 21 (Pd21) and Pd5, the only identified cilia-activating efferent neurons in Tritonia. Second, stimulation of a single DSI elicits beating of the foot cilia in semi-intact preparations and crawling in intact animal treadmill preparations. Third, the DSIs fire at an elevated rate for nearly 1 hr after a swim motor program, which correlates reasonably well with the period freely behaving animals were found to crawl after they swam. Fourth, silencing the tonically active DSIs after a swim motor program substantially reduces or eliminates ongoing cilia neuron firing, indicating that the DSIs are major contributors to the synaptic input driving these cells. Finally, all of the other swim CPG neurons also connect to the cilia neurons, most monosynaptically. Taken together, these observations indicate that the Tritonia swim CPG network participates in producing both escape swimming and crawling. Given the extreme differences between these behaviors---rhythmic versus tonic, muscular versus ciliary, and brief versus prolonged--these findings reveal a striking versatility for a small multifunctional network. PMID- 11880530 TI - Attentional modulation of behavioral performance and neuronal responses in middle temporal and ventral intraparietal areas of macaque monkey. AB - Although many studies have demonstrated that neuronal responses are modulated by attention, the significance of this modulation for behavior is poorly understood. We recorded from neurons in the middle temporal (MT) and ventral intraparietal (VIP) areas in the visual cortex of two macaque monkeys while they performed a motion detection task under two conditions of spatial attention. The ability of the animals to detect the motion was reduced when they withdrew attention from the stimulus. Withdrawing attention also reduced neuronal responses to the motion in both the MT and VIP areas. To compare the neuronal and behavioral effects of attention, the amount of attentional modulation was expressed in units of stimulus strength. On average, attention modulated neuronal responses in MT less than needed to account for the attentional effect on behavior. The opposite was observed in VIP, where the average effect of attention on neuronal responses was greater than that needed to account for behavior. Similar results were obtained when the effects of attention on neuronal response and behavioral performance were compared using a parametric function of stimulus strength. Across neurons in both areas, attentional modulation of neuronal responses was more variable than, and uncorrelated with, attentional modulation of behavioral performance. These findings suggest that attention can alter the average relationship between neuronal activity in visual cortex and behavioral performance. Where this relationship is preserved may indicate which cortical regions are most closely associated with the behavior in a given task. PMID- 11880531 TI - Deletion of CCK2 receptor in mice results in an upregulation of the endogenous opioid system. AB - Stimulation of the brain CCK2 receptor by the C-terminal octapeptide CCK8 of cholecystokinin (CCK) negatively modulates opioid responses. This suggests the existence of physiologically relevant interactions between endogenous CCK and opioid peptides, opening new perspectives particularly in the treatment of pain or drug addiction. CCK2 receptor-deficient mice were used to analyze the incidence of this gene invalidation on opioid system. Compared with wild-type mice, mutants exhibited the following: (1) a hypersensitivity to the locomotor activity induced by inhibitors of enkephalin catabolism or by morphine; (2) a spontaneous hyperalgesia to thermal nociceptive stimulus, which was reversed by previous administration of the NMDA antagonist MK-801 [(+)-5-methyl-10,11-dihydro 5H-dibenzo [a,d] cyclohepten-5,10-imine maleate], and a large reduction in analgesic effects of endogenous or exogenous opioids; and (3) a more severe withdrawal syndrome after chronic morphine treatment. As expected, stimulation of mu, delta, and D2 receptors on brain tissue of wild-type animals induced a dose dependent decrease in adenylate cyclase activity, whereas a striking mirror effect was observed in mutants. All of these results suggest that the absence, in knock-out mice, of the negative feedback control on the opioid system, normally performed out by CCK2 receptor stimulation, results in an upregulation of this system. These biochemical and pharmacological results demonstrate the critical role played by CCK2 receptors in opioid-dependent responses. PMID- 11880532 TI - Local generation of fast ripples in epileptic brain. AB - Aperiodic high-frequency oscillations (>100 Hz) reflect a short-term synchronization of neuronal electrical activity. It has been shown in the epileptic brain that spontaneous oscillations in the frequency range of 250-600 Hz reflect action potential population bursts of synchronously discharging neuronal clusters. These oscillations occur in the early stages of epileptogenesis in areas adjacent to the brain lesion and may trigger the formation of seizure-generating neuronal networks. We studied the extent of the area generating oscillations in the frequency range of 250-600 Hz [fast ripples (FRs)] in intrahippocampal kainic acid-treated rats with spontaneous seizures, by analyzing voltage versus depth profiles of FRs in hippocampal and parahippocampal areas in freely moving animals and by spatial mapping in hippocampal slice preparations in vitro. The strength of inhibition was compared in areas with and without FRs using a paired-pulse paradigm. The extent of the areas generating FRs did not exceed 1 mm(3). The areas generating FRs became broader after the application of the GABA(A) receptor antagonist bicuculline. Paired-pulse fast inhibition at 15-30 msec intervals was similar in areas generating FRs and areas not generating FRs. Our data illustrate that hypothesized clusters of highly interconnected neurons are capable of overcoming interneuron feedback inhibition, resulting in generation of epileptiform bursts, eventually leading to seizure activity. PMID- 11880533 TI - Dominant-negative subunits reveal potassium channel families that contribute to M like potassium currents. AB - M-currents are K+ currents generated by members of the KCNQ family of K+ channels (Wang et al., 1998). However, in some cells, M-like currents may be contaminated by members of other K+ channel gene families, such as the erg family (Meves et al., 1999; Selyanko et al., 1999). In the present experiments, we have used the acute expression of pore-defective mutants of KCNQ3 (DN-KCNQ3) and Merg1a (DN Merg1a) as dominant negatives to separate the contributions of these two families to M-like currents in NG108-15 neuroblastoma hybrid cells and rat sympathetic neurons. Two kinetically and pharmacologically separable components of M-like current could be recorded from NG108-15 cells that were individually suppressed by DN-Merg1a and DN-KCNQ3, respectively. In contrast, only DN-KCNQ3, and not DN Merg1a, reduced currents recorded from sympathetic neurons. Pharmacological tests suggested that the residual current in DN-KCNQ3-treated sympathetic neurons was carried by residual KCNQ channels. Ineffectiveness of DN-Merg1a in sympathetic neurons was not caused by lack of expression, as judged by confocal microscopy of Flag-tagged DN-Merg1a. These results accord with previous inferences regarding the roles of erg and KCNQ channels in generating M-like currents. This experimental approach should therefore be useful in delineating the contributions of members of these two gene families to K+ currents in other cells. PMID- 11880534 TI - P/Q-type calcium-channel blockade in the periaqueductal gray facilitates trigeminal nociception: a functional genetic link for migraine? AB - The discovery of mis-sense mutations in the alpha1A subunit of the P/Q-type calcium channel in patients with familial hemiplegic migraine indicates the potential involvement of dysfunctional ion channels in migraine. The periaqueductal gray (PAG) region of the brainstem modulates craniovascular nociception and, through its role in the descending pain modulation system, may contribute to migraine pathophysiology. In this study we sought to investigate the possible link between the genetic mutations found in migraineurs and the PAG as a modulator of craniovascular nociception. We microinjected the P/Q-type calcium-channel blocker omega-agatoxin IVA into the rat ventrolateral PAG (vlPAG). We examined its effect on the nociceptive transmission of second-order neurons recorded in the trigeminal nucleus caudalis and activated by stimulation of the parietal dura mater. After injection of agatoxin into the vlPAG (n = 20) responses to dural stimulation were facilitated by 143% (p < 0.0001) for Adelta fiber activity and 180% for C-fiber activity (p < 0.05). Similarly, spontaneous background activity increased by 163% (p < 0.0001). These results demonstrate that P/Q-type calcium channels in the PAG play a role in modulating trigeminal nociception and suggest a role for dysfunctional P/Q-type calcium channels in migraine pathophysiology. PMID- 11880535 TI - Variation in effective stimulus patterns for induction of long-term potentiation across different layers of rat entorhinal cortex. AB - Neuronal activities in superficial (II and III) and deep (V and VI) layers of the entorhinal cortex (EC) are preferentially modulated by theta and sharp wave (SPW) EEG, respectively. We investigated the possibility that distinct EEG patterns represent optimal stimulus patterns for induction of long-term potentiation (LTP) in different layers of the EC. We examined effects of three different stimulation patterns on LTP induction in layers I, II-III, and V of medial EC slices of the rat. The stimulation patterns we used were a single, long high-frequency train (1 sec at 100 Hz, repeated 3x), theta burst stimulation [TBS; 10 bursts (four pulses, 100 Hz) at 5 Hz, repeated 3x], and SPW-like burst stimulation [ SPWBS; three bursts (20 pulses, 200 Hz) at 2 Hz, repeated 6x]. Similar degrees of LTP were induced by the three stimulation patterns in layer I. In layers II--III and layer V, however, the largest degrees of LTP were induced by TBS and SPWBS, respectively. These results suggest that burst stimulation constructed to mimic naturally occurring patterns of neuronal activity in the corresponding layer is optimal for LTP induction in layers II-III and layer V of the EC. The differences may play important roles in shaping hippocampal-neocortical interactions in encoding and retrieval of memory. PMID- 11880537 TI - Early induction of genetic instability and apoptosis by arsenic in cultured Chinese hamster cells. AB - In order to assess at what time from the beginning of exposure inorganic arsenic can give rise to genetic instability and trigger apoptosis, V79-C13 Chinese hamster cells were treated with 10 microM sodium arsenite for 24 h. Under these conditions, cell survival was >70% and cells showed neither an increase in chromosome aberration frequency nor a delay in cell cycle progression. Investigations, which were carried out every 6 h during the treatment, revealed an early appearance of genetically unstable cells, namely micronucleated, multinucleated and mononucleated 'giant' cells, as well as apoptotic cells. Indirect immunostaining using anti-beta-tubulin antibody showed severe alterations in spindle morphology after only 6 h treatment, when cells with small spindles whose poles were inside the metaphase plate appeared, and after 12 h treatment, when cells in which spindle assembly had completely failed were observed. These cells, unable to complete mitosis, underwent apoptosis. In fact, cells which turned out to be positive in the TdT-FragEL test had condensed chromatin arranged in metaphase-like plates; their maximum frequency was reached after 24 h treatment. A cytogenetic study was conducted at the end of the period of exposure to arsenic and after post-treatment incubation in fresh medium for up to 5 days. It showed that the percentage of cells with 21 chromosomes (modal number of the cell line) decreased, making way for aneuploid cells. Arsenic, therefore, induced early genetic instability or apoptosis in dividing cells. However, while apoptosis tended to cease when arsenic was removed from the culture medium, the acquired instability remained and propagated within the cell population. PMID- 11880538 TI - Mutant frequencies and loss of heterozygosity induced by N-ethyl-N-nitrosourea in the thymidine kinase gene of L5178Y/TK(+/-)-3.7.2C mouse lymphoma cells. AB - N-ethyl-N-nitrosourea (ENU) is a potent monofunctional ethylating agent that has been found to be mutagenic in a wide variety of organisms from viruses to mammalian germ cells. To elucidate the mutagenicity of ENU at the Tk(+/-) locus of mouse lymphoma cells and to confirm the ability of the mouse lymphoma assay (MLA) to detect both point mutations and large DNA alterations, Tk(+/-) L5178Y cells were exposed to different doses of ENU. Treatment of the cells with ENU resulted in a linear dose response with mutant frequencies of up to 16-fold over control. Evaluation of mutant clone size showed that 36% of the 100 microg/ml ENU induced clones (66% in control) were small colony mutants and 64% (34% in control) were large colony mutants. DNA isolated from mutants in the control culture and the 100 microg/ml ENU treatment group was analyzed for loss of heterozygosity (LOH) using allele-specific PCR. The majority of the small colony mutants, both ENU-treated (97%) and spontaneous (91%), lost the Tk1b allele. The percentage of allele loss in ENU-induced large colony mutants was distinctly different from that of the control. Twenty-three percent of ENU-induced large colony mutants lost their Tk1b alleles, whereas 73% of the large colony mutants from the control culture lost the allele (P < 0.001). Overall, 50% of the Tk mutants from the 100 microg/ml ENU-treated cultures (86% in control) showed LOH. Our data indicate that ENU is a potent mutagen in mouse lymphoma cells and that 100 microg/ml ENU induces equal numbers of point mutations and chromosomal mutations. This study serves to verify that the MLA detects both point mutations and chromosomal mutations. PMID- 11880539 TI - Nature of anaphase laggards and micronuclei in female cytokinesis-blocked lymphocytes. AB - We used pancentromeric fluorescence in situ hybridization and X chromosome painting to characterize late anaphase aberrations in cultured (72 h) female lymphocytes in the presence of cytochalasin B (Cyt-B). Aberrant cells, mostly containing laggards, were very common (34.5%) among multipolar anaphases but fewer (5.4%) among bipolar anaphases. Characterization of the laggards showed that 75% were autosomes, 15% autosomal fragments and 10% X chromosomes in bipolar divisions; similar figures were obtained in multipolar cells. The X chromosome lagged behind more often than would be expected by chance (1/23), representing 12 and 7% of all lagging chromosomes in bipolar and multipolar divisions, respectively. Bipolar divisions contained more lagging autosomes but fewer lagging fragments and X chromosomes with Cyt-B than without it. Comparison of the frequencies of anaphase laggards and interphase micronuclei (MN) showed that lagging autosomes seldom form MN in bipolar divisions, 11% being micronucleated without Cyt-B and 8% with Cyt-B. In multipolar divisions, autosome laggards produced MN more often (35%) and were mainly responsible for the excessive MN frequency of multinucleate cells. Lagging acentric fragments frequently formed MN, with a higher efficiency in the presence of Cyt-B (65% bipolar, 58% multipolar) than in its absence (41%). X chromosome laggards were very easily micronucleated, half of them forming MN in untreated cells and seemingly all after Cyt-B treatment. Our findings suggest that most autosome laggards are merely delayed in their poleward movement, eventually being engulfed by the nucleus. Lagging fragments and X chromosomes are probably detached from the spindle and, therefore, preferentially form MN. X laggards are particularly efficiently micronucleated in Cyt-B-treated cells, perhaps because they stay further away from the poles in round cytokinesis-blocked anaphases than in normally elongated non-blocked anaphases. PMID- 11880540 TI - The CYP17 MspA1 polymorphism and breast cancer risk: a meta-analysis. AB - Inter-individual differences in susceptibility to breast cancer are partially mediated through the levels of endogenous and exogenous steroid hormones. The CYP17 gene encodes P450c17alpha, an enzyme that is involved in the metabolism of steroid hormones. Increased endogenous steroid hormone levels have been associated with a MspA1 polymorphism in the 5'-promoter region of the CYP17 gene. The CYP17 MspA1 polymorphism has been postulated as being associated with the risk of developing breast cancer. However, the association between the CYP17 MspA1 polymorphism and breast cancer risk has been controversial in the literature. To re-examine this controversy, we have undertaken a meta-analysis of 15 case-control studies, which included a total of 4227 breast cancer cases and 4730 individual controls. The odds ratio (OR) was used to evaluate the risk of breast cancer for each study, using homozygosity of the wild-type allele as the control group. Statistical analysis showed no evidence of heterogeneity within the studies. The pooled ORs of breast cancer associated with the combined variant (A1/A2 + A2/A2) and the homozygous genotype (A2/A2) were 0.98 (95% CI 0.89-1.07) and 1.05 (95% CI 0.87-1.21), respectively. Similarly, the pooled ORs of advanced breast cancer associated with the combined variant and the homozygous genotype were 0.96 (95% CI 0.77-1.20) and 0.88 (95% CI 0.55-1.41), respectively. A pooling of the studies was also conducted for the various ethnic groups, but failed to show an association of CYP17 MspA1 polymorphism with breast cancer risk in the different ethnic groups. In addition, our results show that a possible protective effect for breast cancer risk of a later age at menarche was mainly limited to women with the A1 homozygous genotype. The OR for age at menarche (> or = 13) was 0.87 (95% CI 0.62-1.17). Our results suggest that CYP17 MspA1 polymorphism may be at best a weak modifier of breast cancer risk but is not a significant independent risk factor. PMID- 11880541 TI - Indoor and outdoor genotoxic load detected by the Comet assay in leaves of Nicotiana tabacum cultivars Bel B and Bel W3. AB - Environmental pollution assessment and control are priority issues for both developed and developing countries of the world. The use of plant material for a more complete picture of environmental health appears to be particularly appealing. Here we validate a previous plant-adapted Comet assay on leaf tissues of Nicotiana tabacum cultivars Bel B and Bel W3. The effects of H(2)O(2) on DNA damage in Bel B and Bel W3 agree with the hypothesis that some component of the machinery that protects DNA integrity from oxidative stress may be impaired in cv. Bel W3. Exposure in the field on sunny summer days (peak ozone concentration >80 p.p.b.) showed significantly higher DNA damage in cv. Bel W3 if plants were collected and subjected to the Comet assay when the air ozone concentration was reaching its peak value, but not when plants were sampled early in the morning and hence after a period of low ozone concentration. The different results suggest that Bel W3 possesses a less efficient recovery apparatus that requires a longer period of activity to be effective and/or is less protected against reactive oxygen species production during exposure to ozone. However, it cannot be excluded that the increase in mean DNA damage is the result of the presence of a genotoxic agent(s) other than ozone. Interestingly, Bel W3 also appears to be more responsive, compared with Bel B, when exposed to ambient indoor pollutants. The use of cv. Bel W3 increases the sensitivity of the assay under both indoor and field conditions. However, different classes of mutagens should be tested to define the range of profitable utilization of this tobacco cultivar for environmental genotoxicity detection. PMID- 11880542 TI - The frequency of dicentrics and acentrics and the incidence of rogue cells in radiation workers. AB - Occupational exposure to ionizing radiation causes chromosomal damage. Some of the damaged cells show a large number of aberrations such as dicentrics, polycentrics, rings and numerous acentric fragments. This paper describes an analysis of the frequency of dicentric chromosomes and acentric fragments in 1260 subjects occupationally exposed to X-rays and 241 controls. Special attention was paid to the incidence of multi-aberrant cells. The 3 year cumulative dose was a significant predictor for all analyzed aberrations. The duration of exposure was a highly significant predictor of the frequency of rogue cells, but not of acentrics and dicentrics. Age and sex were not found to be significant predictors of the analyzed aberrations. PMID- 11880544 TI - p53 regulation of DNA excision repair pathways. AB - The regulation of DNA excision repair pathways by p53 and its downstream genes is an emerging body of literature, largely distinct and separable from the more studied cell cycle arrest and apoptosis responses regulated by p53. Regulation of nucleotide excision repair of UV-damage by p53 and its downstream genes Gadd45 and p48XPE has been well-documented, but much remains to be done in elucidating mechanisms. Moreover, p53 also participates in base excision repair of hydrogen peroxide-induced damage, still at an early stage of investigation. In human cancers carrying inactivating mutations in p53, especially those wherein p53 mutation occurs early, accelerated mutagenesis by exogenous and endogenous DNA damage is predicted. At the same time, the excision repair pathways could provide a useful target for DNA-damaging chemotherapeutics against p53-defective cancers, having decreased ability to repair chemotherapeutic damage. To our knowledge, this is the first review to address this emerging field. PMID- 11880543 TI - Comparison of camptothecin derivatives presently in clinical trials: genotoxic potency and mitotic recombination. AB - The genotoxicity of camptothecin (CPT) and its clinical antineoplastic analogues irinotecan (CPT-11) and topotecan (TPT) were evaluated using the wing somatic mutation and recombination test (SMART) in Drosophila melanogaster. These compounds stabilize and trap the topoisomerase I-DNA complex, preventing the religation step of the breakage/rejoining reaction mediated by the enzyme. The standard version of the wing SMART was used to evaluate the three compounds and to compare the wing spots induced in marker-heterozygous and balancer heterozygous flies. The results demonstrate that all compounds tested have a significant genotoxic effect in both genotypes analysed. At the same time, a comparison of the clone induction frequencies in marker-heterozygous and balancer heterozygous flies shows that mitotic recombination is the prevalent mechanism through which the three compounds induce all categories of wing spots (78-93% recombination). TPT was the most genotoxic compound, probably because substitutions of amino groups for the 9-carbon of the CPT A ring leads to compounds with greater in vivo activity. CPT and CPT-11 induced, respectively, about 7 and 28 times fewer mutant clones per millimolar exposure unit than TPT. PMID- 11880545 TI - Detection of 1cen--1q12 lesions in different phases of the cell cycle: dual colour FISH analysis of peripheral lymphocytes from subjects with occupational exposure to petroleum fuels. AB - Dual colour FISH was used to assess the genotoxic effects of exposure to petroleum fuels and low benzene levels in peripheral lymphocytes of 12 gasoline station attendants. Labelled DNA probes were used for hybridization of the 1cen and 1q12 contiguous regions of chromosome 1, allowing simultaneous detection of hyperploidy and breakages in both interphase and metaphase cells. The analysis of interphase cells (either unstimulated or mitogen stimulated) showed a prevalence of cells with signal separation in exposed workers compared to matched controls. This difference was highly significant (P < 0.001) in stimulated lymphocytes (9.9 +/- 3.3 and 6.5 +/- 1.5 per thousand in exposed and controls, respectively). Far lower incidences of breaks, with no relation to chemical exposure, were detected in metaphase cells (0.3 +/- 0.8 versus 0.7 +/- 1.0 per thousand, respectively). The analysis of post-mitotic, cytokinesis-blocked cells again showed a relatively high incidence of nuclei with displacement of fluorescent signals (7.2 +/- 2.4 and 5.6 +/- 1.7 per thousand, respectively), suggesting that chromatin decondensation, rather than alteration of DNA strand integrity, led to signal separation in interphase nuclei. Even though the mechanism leading to the separation of alpha and classical satellites in interphase nuclei has not been elucidated, the significant association between cytogenetic findings and intensity of benzene exposure (as shown by the analysis of internal exposure biomarkers) suggests that signal displacement in 1cen-1q12 may be a marker of chemical exposure. PMID- 11880546 TI - A comparison of the in vitro Comet assay with the in vitro chromosome aberration assay using whole human blood or Chinese hamster lung cells: validation study using a range of novel pharmaceuticals. AB - The in vitro Comet assay, a sensitive, quick and relatively cheap test, could become a valid alternative to the commonly used in vitro chromosomal aberration test, in the preliminary evaluation of new chemical entities early in the development of new pharmaceuticals. A validation of the Comet assay procedure using whole human blood or CHL cells was carried out in comparison with a cytogenetic test utilizing the same target cells with the following compounds which demonstrated positive results in standard chromosomal aberration tests: two well-documented clastogens, methyl methanesulphonate and cyclophosphamide, and eight novel drugs in early development. A 3 h exposure time, in both the absence and presence of metabolic activation, was used for the in vitro Comet assay. Agreement between the results of the Comet assay and the chromosomal aberration tests was found to be satisfactory on a qualitative basis, although positive results in the Comet assay were always at higher doses than in the cytogenetic test. This indicates a reduction in sensitivity using the former genotoxicity end point. In order to try to explain this observation, a range of exposure times (0.25, 0.5, 1, 2 and 3 h) were investigated in two further experiments to determine the optimal time for detecting Comet induction in this assay procedure. Maximum levels of DNA damage (in terms of Comet induction) were recorded at earlier sampling times (0.25-1 h) in whole human blood using the same positive doses observed in HPLT. Further studies need to be performed to confirm these findings. It is possible that strand breaks are too short lived to allow detection after a 3 h treatment period (due to preferential repair), indicating the need for shorter exposure times in some cases to optimize their detection. PMID- 11880547 TI - Measurement and analysis of the chemotherapy-induced genetic instability in pediatric cancer patients. AB - Bleomycin sensitivity has been proven to be a useful biomarker for environmental carcinogenesis and tumor genetic instability. We have previously reported a significant increase in the chromosomal aberrations induced by chemotherapy regimens. This study aimed to test whether there is an inherent increased genetic instability in cancer patients at diagnosis, to determine the increase and time course of the chemotherapy-induced instability and to test whether bleomycin sensitivity can be used as a predictor of tumor evolution or relapse. The analysis included 99 pediatric cancer patients with four different tumor types (Ewing's sarcoma, osteosarcoma, lymphoma and CNS tumors) and 25 controls. Blood samples (n = 171) were obtained before and at the end of treatment, during clinical remission and at relapse and bleomycin tests on lymphocyte cultures were performed. We detected a significant increase (P = 0.004) in mutagen sensitivity in patients at the end of treatment compared with untreated patients, regardless of the tumor type. In both the longitudinal and cross-sectional analyses maximal and similar values of mutagen sensitivity were found in patients during treatment (1.84 +/- 0.82) and at relapse (1.78 +/- 0.52); minimum and similar values were found in controls (0.93 +/- 0.23), untreated patients (1.15 +/- 0.65) and in those who had fulfilled the chemotherapy protocols for at least 2 years before their sample was collected (1.09 +/- 0.53). From this preliminary data we can conclude that cytostatic drugs induce a transient increase in chromosomal instability in pediatric cancer patients that can be monitored by bleomycin induced sensitivity tests and that the genetic instability indices should be further investigated as predictors of relapse. PMID- 11880548 TI - Induction of micronuclei in human cell lines and primary cells by combination treatment with gamma-radiation and ethyl methanesulfonate. AB - While testing for genotoxicity is usually performed on single chemicals, exposure of humans often involves combinations of agents. Previous results from this laboratory showed supra-additivity for the induction of micronuclei in p53 mutated mouse lymphoma L5178Y cells after combined treatment with gamma-radiation from a 137Cs source and ethyl methanesulfonate (EMS). The question now was whether supra-additivity was a general phenomenon for the genotoxicity of this combination of a physical and a chemical DNA-damaging agent or whether the result was species- and cell type-specific. The same combination of agents was investigated in two human lymphoblastoid cell lines, TK6 (wild-type p53) and WTK1 (mutated p53), and primary fibroblasts from a fetal human lung. Doses were in the linear dose-effect range, resulting in a 1.5- to 3-fold increase in micronuclei above control. Radiation doses were between 125 and 350 mGy, while the EMS concentrations were 20-50 microg/ml for the cell lines and 250-350 microg/ml for the primary cells. In none of the human test systems was supra-additivity observed. With the WTK1 cells, which are most similar to the mouse cells regarding p53 status, there was even a tendency for a sub-additive combination effect. Possible explanations for the difference to the mouse cells could be related to species-specific aspects, different consequences of the p53 mutations or the presence of additional mutations. It is concluded that caution is advised in the interpretation and extrapolation of experimental results of mixture toxicity data because the outcome could be highly specific for the given selection of agents, doses and assays. PMID- 11880549 TI - Physiological effects of medium-chain triglycerides: potential agents in the prevention of obesity. AB - Medium chain fatty acids (MCFA) are readily oxidized in the liver. Animal and human studies have shown that the fast rate of oxidation of MCFA leads to greater energy expenditure (EE). Most animal studies have also demonstrated that the greater EE with MCFA relative to long-chain fatty acids (LCFA) results in less body weight gain and decreased size of fat depots after several months of consumption. Furthermore, both animal and human trials suggest a greater satiating effect of medium-chain triglycerides (MCT) compared with long-chain triglycerides (LCT). The aim of this review is to evaluate existing data describing the effects of MCT on EE and satiety and determine their potential efficacy as agents in the treatment of human obesity. Animal studies are summarized and human trials more systematically evaluated because the primary focus of this article is to examine the effects of MCT on human energy metabolism and satiety. Hormones including cholescytokinin, peptide YY, gastric inhibitory peptide, neurotensin and pancreatic polypeptide have been proposed to be involved in the mechanism by which MCT may induce satiety; however, the exact mechanisms have not been established. From the literature reviewed, we conclude that MCT increase energy expenditure, may result in faster satiety and facilitate weight control when included in the diet as a replacement for fats containing LCT. PMID- 11880550 TI - Hellen M. Linkswiler (1912-1984). PMID- 11880551 TI - Corn fiber oil lowers plasma cholesterol by altering hepatic cholesterol metabolism and up-regulating LDL receptors in guinea pigs. AB - To evaluate some of the mechanisms involved in the hypocholesterolemic effects of corn fiber oil (CFO), male Hartley guinea pigs were fed diets containing increasing doses of CFO [0 (control), 5, 10 or 15 g/100 g]. Total fat was adjusted to 15 g/100 g in all diets with regular corn oil. Diets contained 0.25 g/100 g cholesterol. A positive control group (LC) with low dietary cholesterol (0.04 g/100 g) was also included. Plasma LDL cholesterol concentrations were 32, 55 and 57% (P < 0.0005) lower with increasing doses of CFO. Compared with controls, intake of CFO resulted in 27-32% lower hepatic microsomal cholesterol (P < 0.0001), the regulatory pool of LDL receptor (LDL-R) expression. CFO intake resulted in favorable plasma and hepatic cholesterol concentrations, similar to those in guinea pigs fed the LC diet. Hepatic cholesterol 7alpha-hydroxylase (CYP7) activity was approximately 88% higher in guinea pigs fed the two higher dosages of CFO (P < 0.05). In parallel, CYP7 mRNA abundance was approximately 88% higher in guinea pigs fed all three CFO diets. CFO treatment also induced hepatic LDLR mRNA by 66-150% with significant differences at the highest CFO dose. These results suggest that CFO, as a result of decreased bile acid absorption, increased mRNA abundance and activity of CYP7. Because hepatic cholesterol is the substrate for CYP7, a lowering of cholesterol concentrations in the total and microsomal pools was observed. As a response to the depleted microsomal free cholesterol pool, the LDL receptor was up-regulated, drawing more cholesterol from plasma, thus leading to the observed decrease in plasma LDL cholesterol concentrations. PMID- 11880552 TI - Catechins from green tea (Camellia sinensis) inhibit bovine and human cartilage proteoglycan and type II collagen degradation in vitro. AB - Polyphenolic compounds from green tea have been shown to reduce inflammation in a murine model of inflammatory arthritis, but no studies have been undertaken to investigate whether these compounds are protective to joint tissues. We therefore investigated the effects of catechins found in green tea on cartilage extracellular matrix components using in vitro model systems. Bovine nasal and metacarpophalangeal cartilage as well as human nondiseased, osteoarthritic and rheumatoid cartilage were cultured with and without reagents known to accelerate cartilage matrix breakdown. Individual catechins were added to the cultures and the amount of released proteoglycan and type II collagen was measured by metachromatic assay and inhibition ELISA, respectively. Possible nonspecific or toxic effects of the catechins were assessed by lactate output and proteoglycan synthesis. Catechins, particularly those containing a gallate ester, were effective at micromolar concentrations at inhibiting proteoglycan and type II collagen breakdown. No toxic effects of the catechins were evident. We conclude that some green tea catechins are chondroprotective and that consumption of green tea may be prophylactic for arthritis and may benefit the arthritis patient by reducing inflammation and slowing cartilage breakdown. Further studies will be required to determine whether these compounds access the joint space in sufficient concentration and in a form capable of providing efficacy in vivo. PMID- 11880553 TI - A high fat meal activates blood coagulation factor VII in rats. AB - In humans, high fat meals cause postprandial activation of blood coagulation factor VII (FVII), but human studies have not provided definite evidence for a prothrombotic effect of dietary FVII activation. An animal model would be an attractive way to pursue this question and therefore we tested the LEW/Mol rat. We gavaged 3 mL of a fat emulsion (n = 42) or 3 mL isotonic glucose (n = 42). Blood was sampled by heart puncture 2, 4 and 6 h (n = 14/group at each time) after the fat/glucose load. Furthermore, blood was sampled from 16 untreated rats to determine the baseline levels. Triglyceride concentrations, activated FVII (FVIIa), FVII coagulant activity (FVIIc), FVII amidolytic activity (FVIIam) and thrombin-antithrombin complexes (TAT) were determined. After fat administration, triglycerides were significantly elevated at 2 h (1.29 mmol/L) and 4 h (1.37 mmol/L) compared with baseline (0.78 mmol/L), and FVIIa was significantly raised at 4 h (54 U/L) and 6 h (58 U/L) compared with baseline (29 U/L). No postprandial changes in FVIIc, FVIIam and TAT were observed. Glucose administration did not affect any variable. We conclude that the LEW/Mol rat is a promising model for use in future studies of thrombotic effects of dietary FVII activation. PMID- 11880554 TI - Dietary iron status has little effect on expression of ceruloplasmin but alters that of ferritin in rats. AB - Evidence supports a role for ceruloplasmin (ferroxidase I) in the release of iron to the blood from mammalian cells. However, recent studies with cultured cells have suggested that it has the opposite effect, and that iron deficiency enhances expression of ceruloplasmin. We therefore examined in rats how nutritional iron status would affect expression of ceruloplasmin. Groups of male Sprague-Dawley rats were reared on a low iron, starch-based diet for 6-8 wk; half were supplemented by injection of iron dextran. At killing, hematocrits of deficient rats were half normal. Supplemented rats had normal liver concentrations of ferritin and ferritin iron. No ferritin was detected in the livers of the deficient rats. Northern analysis showed that ferritin L and H mRNAs were present in the deficient livers, but expression was half that of the normal rats. There was also twice as much copper. Levels of circulating ceruloplasmin (measured by rocket immunoelectrophoresis) were not altered by iron deficiency, although p phenylenediamine oxidase activity and plasma copper were reduced approximately 30%. In repeated studies, no differences in the expression of hepatic ceruloplasmin mRNA were detected. Treatment of rats of both sexes with additional iron (25 mg as iron dextran) 5-14 d before killing increased liver ferritin but did not alter liver ceruloplasmin mRNA expression or levels of circulating ceruloplasmin. We conclude that iron status is not an important factor in the expression of plasma ceruloplasmin made by the liver. However, it does have modest effects on steady-state levels of liver ferritin mRNA. PMID- 11880555 TI - Early postnatal nutrition determines adult pancreatic glucose-responsive insulin secretion and islet gene expression in rats. AB - Human epidemiologic and experimental animal studies suggest strongly that prenatal and early postnatal nutrition influence adult susceptibility to diet related chronic disease. To elucidate biologic mechanisms linking divergent early nutritional sufficiency to adult insulin axis function in an animal model of "metabolic imprinting," this research focused on the following two objectives: 1) identify a tissue responsible for effect persistence, and 2) identify genes showing sustained differential expression in that tissue. Newborn rats were assigned randomly to small (SL), control (C) or large litters (LL) until weaning. Glucose and insulin tolerance tests were conducted directly after weaning (age 26 d) and in adulthood (ages 110 and 255 d). Glucose-stimulated insulin secretion from isolated pancreatic islets was assessed at those ages. DNA microarrays were used to identify genes showing persistent between-group differential expression in isolated islets. Glucose and insulin tolerance tests suggested persistently reduced pancreatic glucose-responsiveness in SL and LL rats. Insulin tolerance tests showed no group differences in whole-body insulin-stimulated glucose uptake. These data support the hypothesis that the endocrine pancreas contributes to primary imprinting in this model. Persistent defects in glucose-stimulated insulin secretion from isolated islets also supported this hypothesis but only in SL rats. Of 13 named islet genes showing SL vs. C differential expression at age 26 d, 10 remained differentially expressed at age 110 d. These data indicate that the endocrine pancreas plays a primary role in the putative metabolic imprinting mechanism in SL rats. PMID- 11880556 TI - Vitamin A and its derivatives induce hepatic glycine N-methyltransferase and hypomethylation of DNA in rats. AB - Regulation of S-adenosylmethionine (SAM) and the SAM/S-adenosylhomocysteine (SAH) ratio by the key cytosolic enzyme glycine N-methyltransferase (GNMT) is essential in optimizing methyl group supply and subsequent functioning of methyltransferase enzymes. Therefore, inappropriate activation of GNMT may lead to the loss of methyl groups vital for many SAM-dependent transmethylation reactions. Previously, we demonstrated that the retinoid derivatives 13-cis- (CRA) and all trans-retinoic acid (ATRA) mediated both the activity of GNMT and its abundance. The present study was conducted to determine whether vitamin A had a similar ability to up-regulate GNMT and to assess the biological importance of GNMT modulation by examining both the transmethylation and transsulfuration pathways after retinoid treatment. Rats were fed a control (10% casein + 0.3% L methionine) diet and orally given retinyl palmitate (RP), CRA, ATRA or vehicle daily for 10 d. RP, CRA and ATRA elevated hepatic GNMT activity 32, 74 and 124%, respectively, compared with the control group. Moreover, the retinoid-mediated changes in GNMT activity were reflected in GNMT abundance (38, 89 and 107% increases for RP-, CRA-, and ATRA-treated rats, respectively). In addition, hepatic DNA, a substrate for SAM-dependent transmethylation, was hypomethylated (approximately 100%) after ATRA treatment compared with the control group. In contrast, the transsulfuration product glutathione was unaffected by retinoid treatment. These results provide evidence of the following: 1) vitamin A, like its retinoic acid derivatives, can induce enzymatically active GNMT; and 2) inappropriate induction of GNMT can lead to a biologically important loss of methyl groups and the subsequent impairment of essential transmethylation processes. PMID- 11880557 TI - The phytoestrogen daidzein affects the antioxidant enzyme system of rat hepatoma H4IIE cells. AB - Phytoestrogens such as the soy isoflavonoid daidzein have potential health benefits. The antioxidant properties of phytoestrogens are considered to be responsible in part for their protective effects. The antioxidant enzyme (AOE) system plays an important role in the defense of cells against oxidative insults. To determine whether flavonoids can exert antioxidative effects not only directly but also indirectly by modulating the AOE system, we investigated the influence of the flavonoid daidzein on the expression of different AOE. Daidzein treatment of hepatoma H4IIE cells increased catalase mRNA expression two- to threefold. Expression levels of copper zinc superoxide dismutase (CuZnSOD) were not affected by exposure to daidzein. Manganese superoxide dismutase (MnSOD) mRNA expression levels decreased slightly and glutathione peroxidase (GPx) levels increased slightly after daidzein exposure. Changes in AOE mRNA expression levels were significant at 300 micromol/L daidzein. To elucidate the mechanisms underlying the strong increase in catalase mRNA, transfection experiments were performed. Transient transfection of hepatoma cells with reporter plasmids containing different parts of the upstream region of the catalase gene showed a significant one- to threefold increase in reporter gene activity after daidzein exposure. This indicates that daidzein can directly activate the rat catalase promoter region. Despite the increase in catalase mRNA, daidzein pretreatment of cells did not protect against oxidative stress resulting from H(2)O(2) exposure. On the contrary, daidzein itself exerted a mild oxidative stress. In conclusion, the changes in the AOE system provoked by daidzein affected the oxidant rather than the antioxidant properties of daidzein. PMID- 11880558 TI - Postruminal administration of partially hydrolyzed starch and casein influences pancreatic alpha-amylase expression in calves. AB - The objective was to examine the effects of postruminal partially hydrolyzed starch (SH) and/or casein on the expression of pancreatic alpha-amylase mRNA, protein and activity in calves. Holstein calves [(n = 24) 88 plus minus 3 kg body weight (BW)], fitted with abomasal infusion cannulas, were randomly assigned within block (week of infusion) to one of four abomasal infusion treatments. Calves were fed an alfalfa-based diet, and SH [4 g/(kg BW.d)] and/or casein [0.6 g/(kg BW.d)] was infused abomasally for 10 d before tissue collection. There was a SH x casein interaction (P < 0.10) for pancreatic weight (g and g/kg BW) because casein increased pancreatic weight in the absence of SH but did not influence pancreatic weight in the presence of SH. Pancreatic alpha-amylase mRNA tended to be lower (P = 0.06) and protein and activity (U/g pancreas and U/g protein) were lower (P = 0.02) in calves receiving abomasal SH. The concentration of pancreatic trypsin activity (U/g pancreas and U/g protein) was lower (P < 0.03) in calves receiving abomasal SH. There was a SH x casein interaction for total alpha-amylase and trypsin activity [U/pancreas and U/(pancreas.kg BW)] because casein increased total activity in the absence of SH but not in the presence of SH. These data suggest that increases in small intestinal protein flow enhance pancreatic weight and thus total pancreatic alpha-amylase and trypsin activity, yet small intestinal SH inhibits the increase in pancreatic weight resulting from increased small intestinal protein flow. Additionally, postruminal SH decreases alpha-amylase expression largely by translational events. PMID- 11880559 TI - Poly(A)(+) RNA encoding proteins capable of transporting L-methionine and/or DL-2 hydroxy-4-(methylthio) butanoic acid are present in the intestinal mucosa of broilers. AB - To investigate the presence of poly(A)(+) RNA that encode proteins capable of transporting L-methionine (L-Met) and/or DL-2-hydroxy-4-(methylthio) butanoic acid (HMB), Xenopus oocytes were injected with poly(A)(+) RNA isolated from broiler intestinal mucosa. Healthy oocytes at stage V or VI were collected from Xenopus laevis and microinjected with water, poly(A)(+) RNA or size-fractioned poly(A)(+) RNA. The ability of the injected oocytes to take up either L-Met or HMB was examined by incubating oocytes with [methyl-(3)H]-L-Met or [5-(14)C]-HMB. A greater uptake of L-Met (P < 0.01) and HMB (P < 0.05) by oocytes injected with poly(A)(+) RNA from the duodenum, jejunum and ileum of the small intestine was observed compared with water-injected oocytes. The greatest (P < 0.05) uptake occurred when poly(A)(+) RNA from the jejunum or ileum was injected. Injections from four different pools of sucrose gradient--fractionated poly(A)(+) RNA from all three intestinal segments induced (P < 0.01) L-Met uptake. There were three to four different pools of sucrose gradient--fractionated poly(A)(+) RNA from the duodenum, jejunum and ileum that induced (P < 0.05) HMB uptake. Uptake of HMB was greater at pH 5.5 than at pH 7.5 and was independent of Na(+). Uptake of L-Met induced by all four poly(A)(+) RNA pools decreased dramatically when Na(+) was removed from the uptake buffer, which indicated that the majority of L-Met uptake was Na(+)-dependent. These results indicate that there are multiple sized poly(A)(+) RNA that encode proteins capable of mediated transport of L-Met and/or HMB present in broiler intestinal mucosa. PMID- 11880560 TI - Molecular cloning and functional expression of a chicken intestinal peptide transporter (cPepT1) in Xenopus oocytes and Chinese hamster ovary cells. AB - To study peptide absorption in chickens, an intestinal peptide transporter cDNA (cPepT1) was isolated from a chicken duodenal cDNA library. The cDNA was 2914 bp long and encoded a protein of 714 amino acid residues with an estimated molecular size of 79.3 kDa and an isoelectric point of 7.48. cPepT1 protein is similar60% identical to PepT1 from rabbits, humans, mice, rats and sheep. Sixteen dipeptides, three tripeptides and four tetrapeptides that contained the essential amino acids Met, Lys and(or) Trp were used for functional analysis of cPepT1 in Xenopus oocytes and Chinese hamster ovary cells. For most di- and tripeptides tested, the substrate affinities were in the micromolar range, indicating that cPepT1 has high affinity for these peptides. Lys-Lys and Lys-Trp-Lys were exceptions, with substrate affinities in the millimolar range. Neither free amino acids nor tetrapeptides were transported by cPepT1. Northern blot analysis using a full-length cPepT1 cDNA as the probe demonstrated that cPepT1 is expressed strongly in the duodenum, jejunum and ileum, and at lower levels in kidney and ceca. The present study demonstrated for the first time the presence and functional characteristics of a peptide transport system from an avian species. PMID- 11880562 TI - Dietary carotenoids contribute to normal human skin color and UV photosensitivity. AB - The aim of the current study was to determine whether dietary carotenoids influence skin pigmentation and UV photosensitivity in a healthy unsupplemented panel (n = 22) of Caucasian (skin Type II) subjects. Skin spectrophotometric and tristimulus (L*a*b*) CR200 chromameter readings were made at various body sites to objectively measure skin carotenoid levels and skin color, respectively. The minimal erythemal dose (MED) was also measured to determine the intrinsic UV photosensitivity of the skin. We found that tristimulus b* values (but not L* and a* values) were consistently and closely correlated with skin carotenoid levels at a number of body sites including the back (r = 0.85, P < 0.00001), forehead (r = 0.85, P < 0.00001), inner forearm (r = 0.75, P < 0.0001) and palm of the hand (r = 0.78, P < 0.0001). Skin carotenoid levels and MED were also correlated in these subjects (r = 0.66, P < 0.001), as were tristimulus b* values and MED (r = 0.71, P < 0.0002). From these observations, we conclude that carotenoids from a normal, unsupplemented diet accumulate in the skin and confer a measurable photoprotective benefit (at least in lightly pigmented Caucasian skin), that is directly linked to their concentration in the tissue. Carotenoids also appear to contribute measurably and significantly to normal human skin color, in particular the appearance of "yellowness" as defined objectively by CR200 tristimulus b* values. On the basis of these findings we believe that objective measurements of skin color, in particular tristimulus b* values, may be a potentially useful means of monitoring dietary carotenoid status and assessing UV photosensitivity in Caucasian populations. PMID- 11880561 TI - Oat consumption does not affect resting casual and ambulatory 24-h arterial blood pressure in men with high-normal blood pressure to stage I hypertension. AB - The results of epidemiologic studies suggest that increased intake of dietary fiber is associated with lower levels of arterial blood pressure (BP). However, there is little information available addressing the possibility that increased oat consumption may reduce arterial BP in individuals with elevated arterial BP. To test this hypothesis, middle-aged and older men (n = 36; body mass index, 25 35 kg/m(2); aged 50-75 y) with elevated BP (systolic BP 130-159 mmHg and/or diastolic BP 85-99 mmHg) were randomly assigned to consume an additional 14 g/d of dietary fiber in the form of oat (5.5 g beta-glucan, n = 18) or wheat cereals (no beta-glucan, n = 18) for 12 wk. Casual resting arterial BP was measured at baseline and after 4, 8 and 12 wk of intervention. The 24-h ambulatory arterial BP was measured at baseline and after 12 wk of intervention. There were no differences in casual resting or 24-h ambulatory BP at baseline in the two groups. Casual systolic BP (SBP) did not change as a result of the 12-wk intervention in the oat (138 plus minus 2 vs. 135 plus minus 3 mmHg) or wheat (142 plus minus 2 vs. 140 plus minus 3 mmHg) groups, respectively (all P > 0.05). Casual diastolic BP (DBP) also did not change in the oat (89 plus minus 2 vs. 88 plus minus 2 mmHg) or wheat (90 plus minus 2 vs. 91 plus minus 2 mmHg) group during this period (all P > 0.05). Further, 24-h, daytime and nighttime SBP and DBP did not decrease with the intervention. Therefore, the results of the present study suggest that any cardioprotective benefit of regular oat consumption may not be conferred via an arterial BP-lowering effect. PMID- 11880563 TI - A food-based formulation provides lycopene with the same bioavailability to humans as that from tomato paste. AB - Lycopene from fresh and unprocessed tomatoes is poorly absorbed by humans. Absorption of lycopene is higher from processed foods such as tomato paste and tomato juice heated in oil. The aim of the present study was to develop a food grade lycopene formulation that is bioavailable in humans. A formulation of lycopene named "lactolycopene" has been designed in which lycopene is entrapped with whey proteins. Healthy subjects (n = 33; 13 men and 20 women) participated and were allocated randomly to one of the three treatment groups. After a 3-wk deprivation of dietary lycopene, subjects ingested 25 mg lycopene/d for 8 wk from lactolycopene, tomato paste (positive control) or a placebo of whey proteins while consuming their self-selected diets. Plasma lycopene concentrations reached a maximum after 2 wk of supplementation in both lycopene-treated groups and then a plateau was maintained until the end of the treatment. Increases in plasma lycopene at wk 8 were not different between supplemented groups (mean +/- SEM): 0.58 +/- 0.13 micromol/L with lactolycopene and 0.47 plus minus 0.07 micromol/L with tomato paste, although they were different from the control (P < 0.001). Similar time-concentration curves of lycopene incorporation were observed in buccal mucosa cells. Although lycopene was present mainly as all-trans isomers (>90%) in both lycopene supplements, plasma lycopene enrichment consisted of 40% as all-trans and 60% as cis isomers. The precursor of lycopene, phytofluene, was better absorbed than lycopene itself. The lactolycopene formulation and tomato paste exhibited similar lycopene bioavailability in plasma and buccal mucosa cells in humans. PMID- 11880564 TI - Olive oil phenols are absorbed in humans. AB - Animal and in vitro studies suggest that olive oil phenols are effective antioxidants. The most abundant phenols in olive oil are the nonpolar oleuropein- and ligstroside-aglycones and the polar hydroxytyrosol and tyrosol. The aim of this study was to gain more insight into the metabolism of those phenols in humans. We measured their absorption in eight healthy ileostomy subjects. We also measured urinary excretion in the ileostomy subjects and in 12 volunteers with a colon. Subjects consumed three different supplements containing 100 mg of olive oil phenols on separate days in random order. Ileostomy subjects consumed a supplement with mainly nonpolar phenols, one with mainly polar phenols and one with the parent compound oleuropein-glycoside. Subjects with a colon consumed a supplement without phenols (placebo) instead of the supplement with oleuropein glycoside. Ileostomy effluent and urine were collected for 24 h after supplement intake. Tyrosol and hydroxytyrosol concentrations were low (< 4 mol/100 mol of intake) in the ileostomy effluent, and no aglycones were detected. We estimated that the apparent absorption of phenols was at least 55-66% of the ingested dose. Absorption was confirmed by the excretion of tyrosol and hydroxytyrosol in urine. In ileostomy subjects, 12 mol/100 mol and in subjects with a colon, 6 mol/100 mol of the phenols from the nonpolar supplement were recovered in urine as tyrosol or hydroxytyrosol. In both subject groups, 5--6 mol/100 mol of the phenols was recovered from the polar supplement. When ileostomy subjects were given oleuropein-glycoside, 16 mol/100 mol was recovered in 24-h urine, mainly in the form of hydroxytyrosol. Thus, humans absorb a large part of ingested olive oil phenols and absorbed olive oil phenols are extensively modified in the body. PMID- 11880565 TI - Once weekly is superior to daily iron supplementation on height gain but not on hematological improvement among schoolchildren in Thailand. AB - Intermittent iron supplementation has been suggested as a replacement for daily iron supplements for reducing anemia in developing countries. The effects of once weekly and daily iron supplementation on hemoglobin (Hb), serum ferritin (SF), prevalence of anemia, weight and height are compared in this study. Primary schoolchildren (n = 397) from two selected schools in the Hat Yai rural area, southern Thailand, were recruited in 1999. All children received Albendazole and then randomly received ferrous sulfate (300 mg/tablet) either daily or weekly, or a placebo for 16 wk. The average increase in Hb was not significantly different between the daily (mean +/- SD; 6.5 +/- 6.0 g/L) and weekly (5.7 +/- 6.3 g/L) groups. However, the average increase in SF was greater (P < 0.01) in the daily (mean +/- SD; 39.8 +/- 30.3 microg/L) than the weekly (13.4 +/- 17.3 microg/L) group. All cases of iron deficiency anemia were abolished in both daily and weekly groups, whereas no reduction in prevalence occurred in the placebo group. Height gain was greater in children who received weekly (mean +/- SD; 2.6 +/- 0.9 cm) than in those who received daily iron (mean +/- SD; 2.3 +/- 0.8 cm), (P < 0.01). Weight gain, weight-for-age and height-for-age were not significantly different among the intervention groups. It is concluded that a weekly iron dose is more effective than a daily dose in height gain but not in hematological improvement over 16 wk of supplementation. PMID- 11880566 TI - Iron deficiency anemia is highly prevalent among human immunodeficiency virus infected and uninfected infants in Uganda. AB - Although anemia is a common finding among human immunodeficiency (HIV)-infected infants in sub-Saharan Africa, the factors contributing to the pathogenesis of anemia have not been well characterized. We sought to characterize the relative contribution of iron deficiency and chronic disease to the anemia among infants. Hemoglobin, ferritin, erythropoietin, tumor necrosis factor-alpha (TNF-alpha), neopterin, CD4(+) lymphocyte count and plasma HIV load were measured in 165 HIV infected and 39 uninfected 9-mo-old infants seen in an outpatient pediatric clinic in Kampala, Uganda. Among HIV-infected and uninfected infants, the prevalence of anemia (hemoglobin < 110 g/L) was 90.9 and 76.9%, respectively (P = 0.015), and the prevalence of iron deficiency anemia (hemoglobin < 110 g/L and ferritin < 12 microg/L) was 44.3 and 45.4%, respectively (P = 0.92). The relatively higher prevalence of anemia among HIV-infected infants was attributed to the anemia of chronic disease. Among infants with and without iron deficiency, the fitted regression line was log(10) plasma erythropoietin = 2.86 - 0.016.hemoglobin, and log(10) plasma erythropoietin = 4.11 - 0.028.hemoglobin, respectively, with a difference in the slope of the regression lines between log(10) erythropoietin and hemoglobin among infants with and without iron deficiency (P = 0.049). Infants in Uganda have an extremely high prevalence of anemia, and nearly half of the anemia is due to iron deficiency. The erythropoietin response to anemia appears to be upregulated among infants with iron deficiency. PMID- 11880567 TI - Tracking of dietary intake patterns of Chinese from childhood to adolescence over a six-year follow-up period. AB - Limited literature has explored whether dietary intake patterns are maintained or changed between childhood and adolescence. More research is required to understand the predictors of these patterns. Using longitudinal data collected in China for 984 children initially aged 6--13 y, we examined children's dietary intake patterns over a 6-y period and the predictors. Detailed data on children's diets (three 24-h recalls in each survey), anthropometry, sociodemographic characteristics and parental education and occupation were collected. To study the predictors of tracking and changes, we conducted ordinary and multinomial logistic regression analyses. Significant correlations (r = 0.28-0.51, P < 0.05) between individuals' 1991 and 1997 dietary intakes were observed for macronutrients and major food groups (vegetable and fruit, meat and edible oil), which were also supported by kappa (a statistic measuring agreement beyond chance). About half of those who initially consumed high fat, high carbohydrate, high vegetable and fruit, and high meat diets continued such diets 6 y later. Family income, urban-rural residence, mother's education and baseline dietary intakes were important predictors of children's dietary intake patterns. In conclusion, even under conditions of rapid socioeconomic change, children are likely to maintain their dietary intake patterns from childhood into adolescence. Efforts to promote healthy eating behaviors may be more effective if focused on younger children, and parents should be involved in these efforts. PMID- 11880568 TI - Assessment of student pharmacists' knowledge concerning folic acid and prevention of birth defects demonstrates a need for further education. AB - Adequate periconceptional consumption of folic acid can prevent neural tube birth defects, and all women capable of becoming pregnant are recommended to consume 400 microg/d. Most women, however, are unaware of this recommendation and do not consume adequate amounts of folic acid. It is important, therefore, that healthcare professionals, such as pharmacists, be capable of educating women regarding folic acid. The aim of this study was to assess knowledge regarding prevention of birth defects by folic acid among student (future) pharmacists in the final year of a professional degree program. Over a 3-y period (1998-2000), students (n = 98) enrolled in a PharmD program completed a survey consisting of five multiple-choice questions concerning folic acid and birth defects. Almost all students (93.9%) correctly identified folic acid as preventing birth defects. Of these students, many also knew that supplementation should begin before pregnancy (73.9%). Fewer, however, were able to correctly identify either the recommended level of intake (55.4%) or good sources of folic acid (57.6-65.2%). These results show that although student (future) pharmacists are aware of folic acid's ability to prevent birth defects, many lack the specific knowledge needed to effectively counsel women in future clinical practice. PMID- 11880570 TI - Trans-10, cis-12 conjugated linoleic acid increases fatty acid oxidation in 3T3 L1 preadipocytes. AB - The purpose of this study was to examine the effect of 0-50 micromol/L trans-10, cis-12 conjugated linoleic acid (CLA) and cis-9, trans-11 CLA isomers on lipid and glucose metabolism in cultures of differentiating 3T3-L1 preadipocytes. Specifically, we investigated the effects of 6 d of CLA treatment on the following: 1) (14)C-glucose and (14)C-oleic acid incorporation and esterification into lipid; 2) (14)C-glucose and (14)C-fatty acid oxidation; and 3) basal and isoproterenol-stimulated lipolysis. Trans-10, cis-12 CLA supplementation (25 and 50 micromol/L) increased both (14)C-glucose and (14)C-oleic acid incorporation into the cellular lipid fraction, which was primarily triglyceride (TG), compared with bovine serum albumin (BSA) controls. Although glucose oxidation ((14)C glucose to (14)C-CO(2)) was unaffected by CLA supplementation, oleic acid oxidation ((14)C-oleic acid to (14)C-CO(2)) was increased by approximately 55% in the presence of 50 micromol/L trans-10, cis-12 CLA compared with BSA controls. In contrast, 50 micromol/L linoleic acid (LA) and cis-9, trans-11 CLA-treated cultures had approximately 50% lower CO(2) production from (14)C-oleic acid compared with control cultures after 6 d of fatty acid exposure. Finally, 50 micromol/L trans-10, cis-12 CLA modestly increased basal, but not isoproterenol stimulated lipolysis compared with control cultures. Thus, the TG-lowering actions of trans-10, cis-12 CLA in cultures of 3T3-L1 preadipocytes may be via increased fatty acid oxidation, which exceeded its stimulatory effects on glucose and oleic acid incorporation into lipid. PMID- 11880569 TI - Alpha-tocopherol metabolism is abnormal in scavenger receptor class B type I (SR BI)-deficient mice. AB - Despite the physiologic importance of vitamin E, in particular its alpha tocopherol (alpha-T) isoform, the molecular mechanisms involved in the cellular uptake of this antioxidant from plasma lipoproteins have not been well-defined. Recent studies have suggested that selective lipid uptake, rather than endocytosis, is important for alpha-T delivery to cells. Here we show that the scavenger receptor class B type I (SR-BI), which mediates cellular selective cholesteryl ester uptake from lipoproteins, facilitates efficient transfer of alpha-T from HDL to cultured cells. In SR-BI-deficient mutant mice, relative to wild-type control animals, there was a significant increase in plasma alpha-T levels (1.1- to 1.4-fold higher) that was mostly due to the elevated alpha-T content of their abnormally large plasma HDL-like particles. This increase in plasma alpha-T in SR-BI knockout mice was accompanied by a significant decrease (65-80%) in the alpha-T concentrations in bile and several tissues including ovary, testis, lung and brain. SR-BI deficiency did not alter the alpha-T concentrations of the liver, spleen, kidney or white fat. These data show that SR BI plays an important role in transferring alpha-T from plasma lipoproteins to specific tissues. Also, in the case of the liver as was previously shown for SR BI-dependent hepatic cholesterol transport, SR-BI-mediated uptake of alpha-T was primarily coupled to biliary excretion rather than to tissue accumulation. Defective tissue uptake of lipoprotein alpha-T in SR-BI-deficient mice may contribute to the reproductive and cardiovascular pathologies exhibited by these animals. PMID- 11880571 TI - Net protein oxidation is adapted to dietary protein intake in domestic cats (Felis silvestris catus). AB - Cats have a requirement for dietary protein two to three times that of omnivores and herbivores. This was reported to be due to the hepatic catabolic enzymes of this species being set to a permanently high level and, therefore, showing little adaptation to low dietary protein. A major mechanism for adapting to dietary protein in other species is amino acid oxidation (hereafter referred to as protein oxidation), and the objective of this study was to determine whether protein oxidation in cats was correlated with protein intake. Net protein and net fat oxidation in six adult cats were studied directly from gas exchanges using indirect calorimetry, after feeding moderate protein (MP; 35% energy) and high protein (HP; 52% energy) diets. Protein oxidation was significantly higher (P < 0.05) when cats were fed the HP diet (28.4 plus minus 0.7 mg/min) rather than the MP diet (20.4 plus minus 0.8 mg/min). Fat oxidation was significantly higher (P < 0.05) when cats consumed the MP diet (9.0 plus minus 0.7 mg/min) rather than the HP diet (4.7 plus minus 0.5 mg/min). Protein oxidation was significantly correlated (linear regression, R(2) = 46.0, P < 0.05) with protein intake such that the mean ratio of 18-h oxidation: 18-h intake was 1.2 on both diets. Fat oxidation was significantly correlated (linear regression, R(2) = 18.9, P < 0.05) with fat intake such that the mean ratio of 18-h fat oxidation: 18-h fat intake was 1.1 (MP) and 0.9 (HP). This study demonstrated that cats adapt net protein oxidation at these levels of protein intake, and the reason for the high dietary protein requirement of this species is, therefore, unclear. PMID- 11880573 TI - Dietary oligofructose and inulin protect mice from enteric and systemic pathogens and tumor inducers. AB - Prebiotics induce changes in the population and metabolic characteristics of the gastrointestinal bacteria, modulate enteric and systemic immune functions, and provide laboratory rodents with resistance to carcinogens that promote colorectal cancer. There is less known about protection from other challenges. Therefore, mice of the B6C3F1 strain were fed for 6 wk a control diet with 100 g/kg cellulose or one of two experimental diets with the cellulose replaced entirely by the nondigestible oligosaccharides (NDO) oligofructose and inulin. From each diet, 25 mice were challenged by a promoter of colorectal cancer (1,2 dimethylhydrazine), B16F10 tumor cells, the enteric pathogen Candida albicans (enterically), or were infected systemically with Listeria monocytogenes or Salmonella typhimurium. The incidences of aberrant crypt foci in the distal colon after exposure to dimethylhdrazine for mice fed inulin (53%) and oligofructose (54%) were lower than in control mice (76%; P < 0.05), but the fructans did not reduce the incidence of lung tumors after injection of the B16F10 tumor cells. Mice fed the diets with fructans had 50% lower densities of C. albicans in the small intestine (P < 0.05). A systemic infection with L. monocytogenes caused nearly 30% mortality among control mice, but none of the mice fed inulin died, with survival intermediate for mice fed oligofructose. Mortality was higher for the systemic infection of S. typhimurium (>80% for control mice), but fewer of the mice fed inulin died (60%; P < 0.05), with mice fed oligofructose again intermediate. The mechanistic basis for the increased resistance provided by dietary NDO was not elucidated, but the findings are consistent with enhanced immune functions in response to changes in the composition and metabolic characteristics of the bacteria resident in the gastrointestinal tract. PMID- 11880572 TI - A systematic screening of total antioxidants in dietary plants. AB - A predominantly plant-based diet reduces the risk for development of several chronic diseases. It is often assumed that antioxidants contribute to this protection, but results from intervention trials with single antioxidants administered as supplements quite consistently do not support any benefit. Because dietary plants contain several hundred different antioxidants, it would be useful to know the total concentration of electron-donating antioxidants (i.e., reductants) in individual items. Such data might be useful in the identification of the most beneficial dietary plants. We have assessed systematically total antioxidants in a variety of dietary plants used worldwide, including various fruits, berries, vegetables, cereals, nuts and pulses. When possible, we analyzed three or more samples of dietary plants from three different geographic regions in the world. Total antioxidants was assessed by the reduction of Fe(3+) to Fe(2+) (i.e., the FRAP assay), which occurred rapidly with all reductants with half-reaction reduction potentials above that of Fe(3+)/Fe(2+). The values, therefore, expressed the corresponding concentration of electron-donating antioxidants. Our results demonstrated that there is more than a 1000-fold difference among total antioxidants in various dietary plants. Plants that contain most antioxidants included members of several families, such as Rosaceae (dog rose, sour cherry, blackberry, strawberry, raspberry), Empetraceae (crowberry), Ericaceae (blueberry), Grossulariaceae (black currant), Juglandaceae (walnut), Asteraceae (sunflower seed), Punicaceae (pomegranate) and Zingiberaceae (ginger). In a Norwegian diet, fruits, berries and cereals contributed 43.6%, 27.1% and 11.7%, respectively, of the total intake of plant antioxidants. Vegetables contributed only 8.9%. The systematic analysis presented here will facilitate research into the nutritional role of the combined effect of antioxidants in dietary plants. PMID- 11880574 TI - Oral administration of arabinogalactan affects immune status and fecal microbial populations in dogs. AB - Seven ileally cannulated dogs were randomly assigned to a control or arabinogalactan (AG) treatments in a 7 x 7 Latin square design to evaluate effects of oral AG administration on nutritional and immunological characteristics. Arabinogalactan treatments included a high (1.65 g/d) and low (0.55 g/d) dose of AG100, AG1000 or AG3000 provided via gelatin capsules. Arabinogalactan forms differed in purification procedures. Each period consisted of a 6-d adaptation followed by a 4-d collection. Blood and fresh fecal samples were collected on d 10 of each period. Fecal score increased (P < 0.02) in dogs supplemented with the low dose of AG1000. Ileal and total tract dry matter (DM) and organic matter (OM) digestibilities were not affected by treatment. Dogs supplemented with the high dose of AG1000 tended (P = 0.15) to have a higher concentration of total aerobic fecal bacteria than control dogs. Dogs supplemented with the low dose of AG1000 and the high dose of AG3000 had higher concentrations of fecal lactobacilli (P = 0.04) and tended to have higher concentrations of fecal bifidobacteria (P < or =0.16) compared with control dogs. Dogs fed the low dose of AG3000 tended (P = 0.10) to have a lower concentration of fecal Clostridium perfringens compared with control dogs. Arabinogalactan treatments did not affect (P > 0.05) serum immunoglobulin G, M or A concentrations. Specific forms and doses of AG increased white blood cell, neutropil and eosinophil concentrations. Arabinogalactan is a unique dietary fiber that affects the digestive physiology and immunological characteristics of dogs. PMID- 11880575 TI - A ketogenic diet increases protein phosphorylation in brain slices of rats. AB - Ketogenic diets have been used to treat seizure disorders of children and recently it was shown to increase the drug-induced seizure threshold in rats. Protein phosphorylation is a major regulatory mechanism of signal transduction that has been implicated in modulating neuronal excitability. We investigated the basal protein phosphorylation in microslices from different brain regions (hippocampus, cerebral cortex and cerebellum) of young rats fed a ketogenic diet, and we evaluated the effect of this diet on weight development and health of these rats based on serum biochemistry. Thirty-day-old rats consumed ad libitum ketogenic (high fat) or control diets for 8 wk. Rats consuming the high fat diet had ketonemia without signs of undernutrition or illness. Microslices were incubated in media containing (32)P-phosphate, and (32)P-phosphoprotein content was analyzed by one- or two-dimensional electrophoresis followed by autoradiography. Basal protein phosphorylation was greater in brain slices from ketogenic rats. Different increments of synapsin I, GAP-43 and GFAP phosphorylation were observed in two-dimensional autoradiography. A ketogenic diet induced metabolic changes affecting the basal status of protein phosphorylation. This change could affect the mechanisms of signal transduction in neural cells involved in the increase in the seizure threshold. PMID- 11880577 TI - Plant breeding: a new tool for fighting micronutrient malnutrition. AB - The final permanent solution to micronutrient malnutrition in developing countries is a substantial improvement in dietary quality--higher consumption of pulses, fruits, vegetables, fish and animal products that the poor already desire but cannot presently afford. Meanwhile breeding staple foods that are dense in minerals and vitamins provides a low-cost, sustainable strategy for reducing levels of micronutrient malnutrition. Getting plants to do the work of fortification, referred to as "biofortification," can reach relatively remote rural populations that conventional interventions are not now reaching and can even have benefits for increased agricultural productivity. Biofortification, thus, complements conventional interventions. The symposium articles discuss several examples of ongoing research projects to develop and disseminate nutrient dense staple food crops and issues that remain to be resolved before successful implementation can be attained. PMID- 11880578 TI - Breeding strategies for biofortified staple plant foods to reduce micronutrient malnutrition globally. AB - One sustainable agricultural approach to reducing micronutrient malnutrition among people at highest risk (i.e., resource-poor women, infants and children) globally is to enrich major staple food crops (e.g., rice, wheat, maize, beans and cassava) with micronutrients through plant-breeding strategies. These target groups are dependent on these staples for their sustenance. Available research has demonstrated that micronutrient-enrichment traits are available within the genomes of these major staple food crops that could allow for substantial increases in the levels of Fe, Zn and provitamin A carotenoids (as well as other nutrients and health-promoting factors) without negatively impacting crop yield. Furthermore, Fe- and Zn-dense seeds can increase crop yields when sowed to soils deficient in these nutrients ensuring their adoption by farmers in these regions once they are developed. Importantly, micronutrient bioavailability issues must be addressed when using a plant-breeding approach to eliminating micronutrient malnutrition. The reduction of antinutrient substances that inhibit micronutrient bioavailability or the increase in substances that promote micronutrient bioavailability from staple plant foods are both options that could be pursued in breeding programs, although care needs to be taken not to compromise agronomic performance and sufficient attention paid to possible beneficial roles of compounds which reduce the bioavailability of trace minerals. The time has come to invest in agricultural technologies to find sustainable solutions to micronutrient malnutrition. Plant breeding is one such technology that should be adopted by the world's agricultural community and that should be supported by the world's nutrition and health communities. PMID- 11880579 TI - Progress in breeding for trace minerals in staple crops. AB - Staples are not considered an important source of minerals in the diet. However, because of high staple consumption, any increase in mineral concentration might well have a significant effect on human nutrition and health. The nutritional quality of staple crops (rice, cassava, wheat, maize and beans) can be improved by breeding. Studies have shown the potential to exploit the genetic variation in seed concentration of iron and other minerals without the general negative effect on yield of adding new traits. The relationship between yield and mineral concentration may be positive, particularly in mineral-deficient soil. Initial evaluations have shown that some crop varieties have high Fe, Zn and carotene in their edible portions. The next step for conventional breeding will be to study the genetics of trace mineral inheritance to determine the best selection technique. Initial investigations of the genetics for high iron in rice have indicated a complex mode of inheritance, demonstrating additive and dominant gene and environmental effects. Breeding strategies have been developed based on these genetic findings. The use of biotechnological tools, such as molecular marker assisted selection, will significantly increase the pace and prospects of success for breeding to improve the nutritional value of staple food crops. PMID- 11880581 TI - Golden Rice: introducing the beta-carotene biosynthesis pathway into rice endosperm by genetic engineering to defeat vitamin A deficiency. AB - To obtain a functioning provitamin A (beta-carotene) biosynthetic pathway in rice endosperm, we introduced in a single, combined transformation effort the cDNA coding for phytoene synthase (psy) and lycopene beta-cyclase (beta-lcy) both from Narcissus pseudonarcissus and both under the control of the endosperm-specific glutelin promoter together with a bacterial phytoene desaturase (crtI, from Erwinia uredovora under constitutive 35S promoter control). This combination covers the requirements for beta-carotene synthesis and, as hoped, yellow beta carotene-bearing rice endosperm was obtained in the T(0)-generation. Additional experiments revealed that the presence of beta-lcy was not necessary, because psy and crtI alone were able to drive beta-carotene synthesis as well as the formation of further downstream xanthophylls. Plausible explanations for this finding are that these downstream enzymes are constitutively expressed in rice endosperm or are induced by the transformation, e.g., by enzymatically formed products. Results using N. pseudonarcissus as a model system led to the development of a hypothesis, our present working model, that trans-lycopene or a trans-lycopene derivative acts as an inductor in a kind of feedback mechanism stimulating endogenous carotenogenic genes. Various institutional arrangements for disseminating Golden Rice to research institutes in developing countries also are discussed. PMID- 11880580 TI - Progress in breeding low phytate crops. AB - Populations that depend on grains and legumes as staple foods consume diets rich in phytic acid (myo-inositol-1,2,3,4,5,6-hexkisphosphate), the storage form of phosphorus in seeds. This compound binds tightly to important mineral nutrients such as iron and zinc, forming salts that are largely excreted. This phenomenon can contribute to mineral depletion and deficiency. As one approach to solving this and environmental problems associated with seed-derived dietary phytic acid, the U. S. Department of Agriculture and others have isolated cereal and legume low-phytic acid mutations and have used these to breed first-generation low phytate hybrids, cultivars and lines of maize (Zea mays), barley (Hordeum vulgare), rice (Oryza sativa) and soybean (Glycine max). Seed phytic acid is reduced in these crops by 50-95%. The progress in the genetics, breeding and nutritional evaluation of low-phytate crops are reviewed in this article. PMID- 11880582 TI - Evaluating the impact of plant biofortification on human nutrition. AB - An evaluation of the efficacy of biofortified foods for improving human nutrition and health requires both laboratory- and community-based trials. A three-step process is proposed. First, tests of nutrient bioavailability should be conducted in the laboratory. Various genotypes of modified foods may be screened for bioavailability using in vitro cell-culture systems or experimental animals before testing in humans. Second, comprehensive feeding trials are conducted to test the efficacy of the biofortified food for improving the nutrition and health of target populations. These trials are generally done for several weeks or months, and they involve measuring a comprehensive set of endpoints. If efficacy is demonstrated in the feeding trial, the third step, a community-based trial, is planned. This final trial involves evaluating the nutritional, health, agricultural, societal, environmental and economic effects of the biofortified food in the community. A multidisciplinary team including consumers, policymakers, health leaders, as well as scientists is required for successful completion of the community trial. PMID- 11880583 TI - Transgenic approaches in commonly consumed cereals to improve iron and zinc content and bioavailability. AB - Modern genetic and molecular technologies provide a number of tools that can be utilized for the development of staple foods with a higher iron and zinc content and improved bioavailability of these minerals. This article summarizes current strategies aimed at increasing the iron-sequestering capacity of the endosperm and improving mineral bioavailability via in planta synthesis of microbial phytases. A case study is presented for wheat, and future strategies are discussed addressing the importance of phytase thermostability. PMID- 11880585 TI - The body of evidence to support a protective role for lutein and zeaxanthin in delaying chronic disease. Overview. AB - Recent evidence introduces the possibility that lutein and zeaxanthin may protect against the development of the two common eye diseases of aging, cataract and macular degeneration. This potential and the lack of other effective means to slow the progression of macular degeneration have fueled high public interest in the health benefits of lutein and zeaxanthin and the proliferation of supplements containing them on pharmacy shelves. An understanding of the biologic consequences of limiting or supplementing with these carotenoids is only beginning to emerge. Some epidemiologic evidence supports a role in eye disease and, to a lesser extent, cancer and cardiovascular disease. However, the overall body of evidence is insufficient to conclude that increasing levels of lutein and zeaxanthin, specifically, will confer an important health benefit. Future advances in scientific research are required to gain a better understanding of the biologic mechanisms of their possible role in preventing disease. Additional research is also required to understand the effect of their consumption, independent of other nutrients in fruits and vegetables, on human health. The newly advanced ability to measure levels of lutein and zeaxanthin in the retina in vivo creates a unique opportunity to contribute some of this needed evidence. PMID- 11880587 TI - Factors that influence the bioavailablity of xanthophylls. PMID- 11880586 TI - Diet and lifestyle correlates of lutein in the blood and diet. AB - Observational studies have suggested an inverse relationship between dietary or serum lutein and risk for age-related macular degeneration and cataracts. This evidence has stimulated interest in the biological and other characteristics of lutein, and also zeaxanthin, a structurally similar carotenoid; together, they comprise the macular pigment. Accurate interpretation of data linking dietary intake or serum concentrations of lutein and zeaxanthin and risk for eye disease in epidemiologic and clinical studies requires knowledge of biological and nondietary factors influencing these intake data or concentrations. The primary aims of this study were to identify the correlates of dietary lutein + zeaxanthin intake and the determinants of serum lutein and zeaxanthin concentrations in a heterogeneous community-based sample of adults aged 18-92 y, recruited and examined at three U.S. sites (n = 2786). An additional aim was to identify the determinants of change in serum lutein concentration from baseline to 1 y in a subset of 1368 study participants followed prospectively. Demographic characteristics (age, sex, race/ethnicity, education), body mass index and lifestyle factors (exercise, sun exposure, smoking, alcohol consumption) were significantly (P < 0.05) associated with dietary lutein + zeaxanthin intake. Demographic characteristics, dietary intake, serum cholesterol concentration, body mass index and smoking explained 24% of the variance in serum lutein concentration. Race/ethnicity, education level and smoking had the strongest associations with serum lutein concentration. Every 10% increase in dietary lutein + zeaxanthin intake was associated with a 2.4% increase in serum lutein concentration. Notably, however, the amount of variance in serum concentration that is explained by demographic characteristics, health-related behaviors and lifestyle factors remains substantial. PMID- 11880588 TI - In vivo assessment of retinal carotenoids: macular pigment detection techniques and their impact on monitoring pigment status. AB - Of the many carotenoids found within human tissue, only the carotenoids within the human retina can be assessed noninvasively at present. Such assessment should eventually provide a more complete understanding of the functional role of retinal lutein (L) and zeaxanthin (Z) (termed macular pigment, MP) in human vision. The emerging data allow for some initial observations. For example, there appears to be wide variation (>factor of 10) in the concentration of MP. Although MP levels have been recorded from nondetectable to 1.20 OD (optical density), the "average" levels, relative to what is possible, appear low. This may be due in part to the low average dietary intake of L and Z in the typical U.S. diet. Nonetheless, individual differences in MP may also be influenced by nondietary factors such as genetics, demographics and lifestyle characteristics. Some evidence indicates that the MP carotenoids may protect the retina and lens, and could improve vision through some optical mechanisms. Consequently, efforts to determine typical MP levels and the factors that influence individual differences in MP density should be continued. PMID- 11880589 TI - Possible biologic mechanisms for a protective role of xanthophylls. AB - This contribution surveys the evidence linking the presence of the two xanthophylls, lutein and zeaxanthin, to a protective role in the macular region of the retina. Although the evidence is still associative in nature, it is biologically plausible, and may be resolved with additional intervention trials. PMID- 11880590 TI - Introduction and satellite session at the fourth International Symposium on the Role of Soy in Preventing and Treating Chronic Disease. PMID- 11880591 TI - Gaining insight into the health effects of soy but a long way still to go: commentary on the fourth International Symposium on the Role of Soy in Preventing and Treating Chronic Disease. AB - Research into the health effects of soyfoods and soybean constituents has increased at a phenomenal pace over the past decade. This research includes a wide range of areas, such as cancer, coronary heart disease, osteoporosis, cognitive function, menopausal symptoms and renal function. Importantly, there are an increasing number of clinical studies being conducted in this field, which was quite evident from the findings presented at the Fourth International Symposium on the Role of Soy in Preventing and Treating Chronic Disease, November 4-7, 2001, in San Diego, California. There is no doubt that progress in understanding the health effects of soy is being made, but much of the data are frustratingly inconsistent. For example, there were conflicting results presented at the symposium on the role of isoflavones in bone health. Similarly, presentations painted an unclear picture of the role of isoflavones in cholesterol reduction. The relatively short duration and small sample size of many of the human studies in this field likely contribute to the inconsistent results. Although there are some controversies regarding the safety of soy for certain subsets of the population, special sessions at the symposium on breast cancer and cognitive function did much to alleviate concerns that soy could have detrimental effects in these areas. Furthermore, published data and new research presented at this meeting suggest that the consumption of even 10 g (typical of Asian intake) of isoflavone-rich soy protein per day may be associated with health benefits. If this modest amount of soy protein were to be incorporated in the American diet, it would represent only approximately 15% of total U. S. protein intake. PMID- 11880592 TI - Genistein chemoprevention: timing and mechanisms of action in murine mammary and prostate. AB - We investigated the potential of genistein, the primary isoflavone of soy, to protect against breast and prostate cancers in animal models. For mammary cancer studies, Sprague-Dawley rats were fed AIN-76A diet plus minus 250 mg genistein/kg diet. Dimethylbenz[a]anthracene was administered by gavage at d 50 postpartum to induce mammary tumors. Mammary cancer chemoprevention was demonstrated after prepubertal and combined prepubertal and adult genistein treatments but not after prenatal- or adult-only treatments, demonstrating that the timing of exposure to genistein is important for mammary cancer chemoprevention. The cellular mechanism of action was found to be mammary gland and cell differentiation, as shown by whole-mount analysis and beta-casein expression. An imprinting effect was shown for epidermal growth factor receptor expression in mammary terminal end buds. For prostate cancer studies, we used two models. The first was a chemically (N methylnitrosourea) induced prostate cancer rat model. Genistein in the diet inhibited the development of invasive adenocarcinomas in a dose-dependent manner. The second model was a transgenic mouse model that resulted in spontaneously developing adenocarcinoma tumor of the prostate. Genistein in the diet reduced the incidence of poorly differentiated prostatic adenocarcinomas in a dose dependent manner and down-regulated androgen receptor, estrogen receptor-alpha, progesterone receptor, epidermal growth factor receptor, insulin-like growth factor-I, and extracellular signal-regulated kinase-1 but not estrogen receptor beta and transforming growth factor-alpha mRNA expressions. We conclude that dietary genistein protects against mammary and prostate cancers by regulating specific sex steroid receptors and growth factor signaling pathways. PMID- 11880594 TI - Soy, soy phytoestrogens and cardiovascular disease. AB - Dietary soy protein has been shown to have several beneficial effects on cardiovascular health. The best-documented effect is on plasma lipid and lipoprotein concentrations, with reductions of approximately 10% in LDL cholesterol concentrations (somewhat greater for individuals with high pretreatment LDL cholesterol concentrations) and small increases in HDL cholesterol concentrations. Dietary soy protein improves flow-mediated arterial dilation of postmenopausal women but worsens that of men. Soy isoflavone extracts improve systemic arterial compliance, an indicator of atherosclerosis extent. Complete soy protein but not alcohol-washed soy protein reduces atherosclerosis of postmenopausal monkeys. No definite experimental evidence exists currently to establish that the cardiovascular benefits of soy protein are accounted for by its isoflavones. PMID- 11880593 TI - The health consequences of early soy consumption. AB - Infants fed soy formula are the segment of the U. S. population that consumes the most soy. Before birth and after weaning, most Americans are not exposed to appreciable levels of soyfoods other than foods that have small amounts of processed soy components. The opposite scenario occurs in Asia, because Asians are more likely to consume relatively high levels of soyfoods throughout life, except between birth and weaning, when breastfeeding or milk-based formula are common. Soy formula is made with soy protein isolate containing isoflavones (SPI+) and supports normal growth and development in term infants. Recent data suggest that there are no long-term adverse effects of early exposure to soy formula through young adulthood. It is as yet unknown whether soy formula consumption by infants will result in health problems or benefits upon aging, but multigenerational animal studies with diets made with SPI+ have not revealed any problems. Soy isoflavones can function as estrogen agonists, antagonists or selective estrogen receptor modulators, depending on the conditions, and much research has focused on health effects of purified isoflavones. Results from several studies suggest that the effects of diets made with SPI+ differ significantly from those of diets to which purified soy isoflavones are added. Furthermore, it seems that soy protein processed to contain lower levels of isoflavones also provides significant health benefits. Further research is needed to confirm the results of the few studies that have been conducted and new studies are needed to investigate the more subtle effects that could occur during development or that could surface later in life. PMID- 11880595 TI - Hormonal effects of soy in premenopausal women and men. AB - Over the past few years, there has been increasing interest in the possible hormonal effects of soy and soy isoflavone consumption in both women and men. Soy consumption has been suggested to exert potentially cancer-preventive effects in premenopausal women, such as increased menstrual cycle length and sex hormone binding globulin levels and decreased estrogen levels. There has been some concern that consumption of phytoestrogens might exert adverse effects on men's fertility, such as lowered testosterone levels and semen quality. The studies in women have provided modest support for beneficial effects. One cross-sectional study showed serum estrogens to be inversely associated with soy intake. Seven soy intervention studies controlled for phase of menstrual cycle. These studies provided 32-200 mg/d of isoflavones and generally showed decreased midcycle plasma gonadotropins and trends toward increased menstrual cycle length and decreased blood concentrations of estradiol, progesterone and sex hormone-binding globulin. A few studies also showed decreased urinary estrogens and increased ratios of urinary 2-(OH) to 16alpha-(OH) and 2-(OH) to 4-(OH) estrogens. Soy and isoflavone consumption does not seem to affect the endometrium in premenopausal women, although there have been weak estrogenic effects reported in the breast. Thus, studies in women have mostly been consistent with beneficial effects, although the magnitude of the effects is quite small and of uncertain significance. Only three intervention studies reported hormonal effects of soy isoflavones in men. These recent studies in men consuming soyfoods or supplements containing 40--70 mg/d of soy isoflavones showed few effects on plasma hormones or semen quality. These data do not support concerns about effects on reproductive hormones and semen quality. PMID- 11880598 TI - Designing supramolecular porphyrin arrays that self-organize into nanoscale optical and magnetic materials. AB - Tessellation of nine free-base porphyrins into a 3 x 3 array is accomplished by the self-assembly of 21 molecular entities of four different kinds, one central, four corner, and four side porphyrins with 12 trans Pd(II) complexes, by specifically designed and targeted intermolecular interactions. Strikingly, the self-assembly of 30 components into a metalloporphyrin nonamer results from the addition of nine equivalents of a first-row transition metal to the above milieu. In this case each porphyrin in the nonameric array coordinates the same metal such as Mn(II), Ni(II), Co(II), or Zn(II). This feat is accomplished by taking advantage of the highly selective porphyrin complexation kinetics and thermodynamics for different metals. In a second, hierarchical self-assembly process, nonspecific intermolecular interactions can be exploited to form nanoscaled three-dimensional aggregates of the supramolecular porphyrin arrays. In solution, the size of the nanoscaled aggregate can be directed by fine-tuning the properties of the component macrocycles, by choice of metalloporphyrin, and the kinetics of the secondary self-assembly process. As precursors to device formation, nanoscale structures of the porphyrin arrays and aggregates of controlled size may be deposited on surfaces. Atomic force microscopy and scanning tunneling microscopy of these materials show that the choice of surface (gold, mica, glass, etc.) may be used to modulate the aggregate size and thus its photophysical properties. Once on the surface the materials are extremely robust. PMID- 11880599 TI - Self-assembly of nanoscopic coordination cages of D(3h) symmetry. AB - A family of nanoscale-sized supramolecular cage compounds with a trigonal prismatic framework was prepared by means of spontaneous self-assembly from the combination of a predesigned molecular "clip" with tritopic pyridyl subunits. As confirmed by x-ray crystallography, the smallest structure of the reported series is approximately 1 x 2 nm and possesses a nitrate anion incarcerated inside its molecular cavity. The largest structure has dimensions of approximately 1 x 4 nm. PMID- 11880600 TI - Molecular dynamics analysis of a buckyball-antibody complex. AB - This is a multinanosecond molecular dynamics study of a bio-nano complex formed by a carbon nanoparticle, a buckyball C(60), and a biological molecule, an antibody, with high binding affinity and specificity. In the simulation, the ball is completely desolvated by the binding site of the antibody by means of a nearly perfect shape complementarity and extensive side-chain interactions, with the exception that about 17% of the surface is persistently exposed to solvent and could be used for functional derivatization. The interactions are predominantly hydrophobic, but significant polar interactions occur as well. There exists a rich body of various pi-stacking interactions. Aromatic side chains are involved in both double and triple stackings with the ball. Some ionic side chains, such as the guanidinium group of arginine, also form pi-stackings with the ball. The results suggest that pi-stackings are very efficient and common modes of biological recognition of pi-electron-rich carbon nanoparticles. Most importantly, the results demonstrate that, in general, an ordinary protein binding site, such as that of an antibody, can readily bind to a carbon nanoparticle with high affinity and specificity through recognition modes that are common in protein-ligand recognition. PMID- 11880601 TI - Infinite pleated beta -sheet formed by the beta-hairpin Boc-beta-Phe-beta-Phe-D Pro-Gly-beta-Phe-beta-Phe-OMe. AB - A beta-hairpin conformation and extended beta-pleated sheet assembly have been characterized by single crystal x-ray diffraction for the synthetic peptide t butoxycarbonyl--beta-Phe-beta-Phe-D-Pro-Gly-beta-Phe-beta-Phe-methyl ester [beta Phe: (S)-beta(3) homophenylalanine]. The centrally located D-Pro-Gly segment nucleates a chain reversal in a type II' beta-turn conformation. Two intramolecular cross-strand hydrogen bonds stabilize the peptide fold. Intermolecular NH...O[double bond]C hydrogen bonds (two on each side of the hairpin) connect the hairpins into an infinitely extended beta-sheet. The beta residues cause all C[double bond]O groups to point in the same direction, resulting in a "polar" sheet by the unidirectional alignment of NH...O[double bond]C hydrogen bonds. In contrast, beta-sheets formed by alpha-residues have alternating directions for the hydrogen bonds, thus resulting in an "apolar" sheet. The crystallographic parameters for C(53)H(66)N(6)O(9) x CH(3)OH are: space group P2(1), a = 9.854(2) A, b = 10.643(2) A, c = 25.296(4) A, beta = 100.39(2) degrees, Z = 2, agreement factor R(1) = 0.065 for 3,706 data observed >4 sigma(F) and a resolution of 0.90 A. PMID- 11880602 TI - Supramolecular templating in thermodynamically controlled synthesis. PMID- 11880603 TI - Supramolecular organic photochemistry: control of covalent bond formation through noncovalent supramolecular interactions and magnetic effects. AB - Supramolecular organic photochemistry, a field concerned with the interaction of light with supramolecular assemblies of organic molecules, has been inspired by the remarkable structural and dynamic features of guest@host chemistry, particularly as exemplified by enzymes. Exemplars of supramolecular organic photochemistry from soft-matter hosts (micelles) and hard-matter hosts (porous solids) are discussed with an emphasis on how noncovalent interactions, which are at the heart of supramolecular chemistry, can be systematically exploited to control the catalytic and magnetic effects on the formation of covalent bonds from photochemically produced pairs of radicals. PMID- 11880604 TI - Molecules within molecules: recognition through self-assembly. AB - Synthetic molecular receptors that completely surround their target molecules can be created through the use of noncovalent interactions. These molecular capsules selectively sequester guest molecules from the influence of bulk solvent and other molecules on the basis of size, shape, and chemical complementarity. This reversible isolation spawns unique behavior within the confines of the host; the catalysis of chemical reactions and the stabilization of reactive species are possible outcomes that have been recently demonstrated. Compartmentalization of reagents can also have a dramatic effect on reactions that take place outside of the capsule, producing nonlinear kinetics in relatively simple reaction systems. PMID- 11880606 TI - Supramolecular assembly dynamics. PMID- 11880605 TI - On the sequencing of the human genome. AB - Two recent papers using different approaches reported draft sequences of the human genome. The international Human Genome Project (HGP) used the hierarchical shotgun approach, whereas Celera Genomics adopted the whole-genome shotgun (WGS) approach. Here, we analyze whether the latter paper provides a meaningful test of the WGS approach on a mammalian genome. In the Celera paper, the authors did not analyze their own WGS data. Instead, they decomposed the HGP's assembled sequence into a "perfect tiling path", combined it with their WGS data, and assembled the merged data set. To study the implications of this approach, we perform computational analysis and find that a perfect tiling path with 2-fold coverage is sufficient to recover virtually the entirety of a genome assembly. We also examine the manner in which the assembly was anchored to the human genome and conclude that the process primarily depended on the HGP's sequence-tagged site maps, BAC maps, and clone-based sequences. Our analysis indicates that the Celera paper provides neither a meaningful test of the WGS approach nor an independent sequence of the human genome. Our analysis does not imply that a WGS approach could not be successfully applied to assemble a draft sequence of a large mammalian genome, but merely that the Celera paper does not provide such evidence. PMID- 11880607 TI - Study of Nd3+, Pd2+, Pt4+, and Fe3+ dopant effect on photoreactivity of TiO2 nanoparticles. AB - The metallorganic chemical vapor deposition method was successfully used to synthesize pure TiO(2) and Nd(3+)-, Pd(2+)-, Pt(4+)-, and Fe(3+)-doped TiO(2) nanoparticles. Polycrystalline TiO(2) structure was verified with x-ray diffraction, which showed typical characteristic anatase reflections without any separate dopant-related peaks. Transmission electron microscopy observations confirmed the existence of homogeneously distributed 22 +/- 3 nm TiO(2) nanoparticles. The particle size remained the same for the doped samples. The doping level of transition metals was kept at approximately 1 atomic percent, which was determined by x-ray photoelectron spectra and energy dispersive x-ray spectroscopy. The effects of different types of dopants on the photocatalytic activity were revealed by the degradation of 2-chlorophenols with an UV light source. The photocatalytic efficiency was remarkably enhanced by the introduction of Pd(2+) and Nd(3+). Nd(3+)-doped TiO(2) showed the largest enhancement. However, Pt(4+) changed the 2-chlorophenol degradation rate only slightly, and Fe(3+) was detrimental to this process. These effects were related to the position of the dopants in the nanoparticles and the difference in their ionic radii with respect to that of Ti(4+). PMID- 11880608 TI - A mitochondrial-like aconitase in the bacterium Bacteroides fragilis: implications for the evolution of the mitochondrial Krebs cycle. AB - Aconitase and isocitrate dehydrogenase (IDH) enzyme activities were detected in anaerobically prepared cell extracts of the obligate anaerobe Bacteroides fragilis. The aconitase gene was located upstream of the genes encoding the other two components of the oxidative branch of the Krebs cycle, IDH and citrate synthase. Mutational analysis indicates that these genes are cotranscribed. A nonpolar in-frame deletion of the acnA gene that encodes the aconitase prevented growth in glucose minimal medium unless heme or succinate was added to the medium. These results imply that B. fragilis has two pathways for alpha ketoglutarate biosynthesis-one from isocitrate and the other from succinate. Homology searches indicated that the B. fragilis aconitase is most closely related to aconitases of two other Cytophaga-Flavobacterium-Bacteroides (CFB) group bacteria, Cytophaga hutchinsonii and Fibrobacter succinogenes. Phylogenetic analysis indicates that the CFB group aconitases are most closely related to mitochondrial aconitases. In addition, the IDH of C. hutchinsonii was found to be most closely related to the mitochondrial/cytosolic IDH-2 group of eukaryotic organisms. These data suggest a common origin for these Krebs cycle enzymes in mitochondria and CFB group bacteria. PMID- 11880609 TI - Emulating biology: building nanostructures from the bottom up. AB - The biological approach to nanotechnology has produced self-assembled objects, arrays and devices; likewise, it has achieved the recognition of inorganic systems and the control of their growth. Can these approaches now be integrated to produce useful systems? PMID- 11880610 TI - Energetics of nanocrystalline TiO2. AB - The energetics of the TiO(2) polymorphs (rutile, anatase, and brookite) were studied by high temperature oxide melt drop solution calorimetry. Relative to bulk rutile, bulk brookite is 0.71 +/- 0.38 kJ/mol (6) and bulk anatase is 2.61 +/- 0.41 kJ/mol higher in enthalpy. The surface enthalpies of rutile, brookite, and anatase are 2.2 +/- 0.2 J/m(2), 1.0 +/- 0.2 J/m(2), and 0.4 +/- 0.1 J/m(2), respectively. The closely balanced energetics directly confirm the crossover in stability of nanophase polymorphs inferred by Zhang and Banfield (7). An amorphous sample with surface area of 34,600 m(2)/mol is 24.25 +/- 0.88 kJ/mol higher in enthalpy than bulk rutile. PMID- 11880611 TI - Quantum dot artificial solids: understanding the static and dynamic role of size and packing disorder. AB - This perspective examines quantum dot (QD) superlattices as model systems for achieving a general understanding of the electronic structure of solids and devices built from nanoscale components. QD arrays are artificial two-dimensional solids, with novel optical and electric properties, which can be experimentally tuned. The control of the properties is primarily by means of the selection of the composition and size of the individual QDs and secondly, through their packing. The freedom of the architectural design is constrained by nature insisting on diversity. Even the best synthesis and separation methods do not yield dots of exactly the same size nor is the packing in the self-assembled array perfectly regular. A series of experiments, using both spectroscopic and electrical probes, has characterized the effects of disorder for arrays of metallic dots. We review these results and the corresponding theory. In particular, we discuss temperature-dependent transport experiments as the next step in the characterization of these arrays. PMID- 11880612 TI - A cut above: discovery of an alternative excision repair pathway in bacteria. PMID- 11880614 TI - Chemical amplification with encapsulated reagents. AB - Autocatalysis and chemical amplification are characteristic properties of living systems, and they give rise to behaviors such as increased sensitivity, responsiveness, and self-replication. Here we report a synthetic system in which a unique form of compartmentalization leads to nonlinear, autocatalytic behavior. The compartment is a reversibly formed capsule in which a reagent is sequestered. Reaction products displace the reagent from the capsule into solution and the reaction rate is accelerated. The resulting self-regulation is sensitive to the highly selective molecular recognition properties of the capsule. PMID- 11880613 TI - Induction of T cell alertness by bacterial colonization of intestinal epithelium. PMID- 11880615 TI - Asymptotic solution of the cylindrical nonlinear Poisson-Boltzmann equation at low salt concentration: analytic expressions for surface potential and preferential interaction coefficient. AB - The analytic solution to the nonlinear Poisson-Boltzmann equation describing the ion distributions surrounding a nucleic acid or other cylindrical polyions as a function of polyion structural quantities and salt concentration ([salt]) has been sought for more than 80 years to predict the effect of these quantities on the thermodynamics of polyion processes. Here we report an accurate asymptotic solution of the cylindrical nonlinear Poisson-Boltzmann equation at low to moderate concentration of a symmetrical electrolyte (< or = 0.1 M 1:1 salt). The approximate solution for the potential is derived as an asymptotic series in the small parameter var epsilon(-1), where var epsilon identical with kappa(-1)/a, the ratio of the Debye length (kappa(-1)) to the polyion radius (a). From the potential at the polyion surface, we obtain the coulombic contribution to the salt-polyelectrolyte preferential interaction (Donnan) coefficient (Gamma(u)coul) per polyion charge at any reduced axial charge density xi. Gamma(u)coul is the sum of the previously recognized low-salt limiting value and a salt-dependent contribution, analytically derived here in the range of low-salt concentrations. As an example of the application of this solution, we obtain an analytic expression for the derivative of the midpoint temperature of a nucleic acid conformational transition with respect to the logarithm of salt concentration (dT(m)/d ln[salt]) in terms of [salt] and nucleic acid structural quantities. This expression explains the experimental observation that this derivative is relatively independent of salt concentration but deviates significantly from its low-salt limiting value in the range 0.01-0.1 M. PMID- 11880616 TI - Matching gene activity with physiological functions. AB - Matching the activity of the genes with biomechanics and physiology is an effective way to use cDNA microarray technology. Required are data on the change of activities of genes associated with specific physiological functions with respect to a continuous variable such as time. For each pair of data (gene and physiological function) as functions of time, we can compute a coefficient of correlation, R. The correlation is perfect if R is +1 or -1; it is nonexistent if R = 0. By evaluating R for every gene in a microarray, we can arrange the genes in the order of the number R, thus learning which genes are best correlated with the mechanical or physiological function. We illustrate this procedure by studying the blood vessels in the lung in response to pulmonary hypoxic hypertension, including the remodeling of vascular morphometry, the elastic moduli, and the zero-stress state of the vessel wall. For each physiological function, we identify the top genes that correlate the best. We found that different genes correlate best with a given function in large and small arteries, and that the genes in pulmonary veins which respond to arterial functions are different from those in pulmonary arteries. We found one set of genes matching the remodeling of arterial wall thickness, but another set of genes whose integral of activity over time best fit the wall thickness change. Our method can be used to study other thought-provoking problems. PMID- 11880618 TI - Mechanism for antibody catalysis of the oxidation of water by singlet dioxygen. AB - Wentworth et al. [Wentworth, P., Jones, L. H., Wentworth, A. D., Zhu, X. Y., Larsen, N. A., Wilson, I. A., Xu, X., Goddard, W. A., Janda, K. D., Eschenmoser, A. & Lerner, R. A. (2001) Science 293, 1806-1811] recently reported the surprising result that antibodies and T cell receptors efficiently catalyze the conversion of molecular singlet oxygen (1O2) plus water to hydrogen peroxide (HOOH). Recently, quantum mechanical calculations were used to delineate a plausible mechanism, involving reaction of 1O2 with two waters to form HOOOH (plus H2O), followed by formation of HOOOH dimer, which rearranges to form HOO HOOO + H2O, which rearranges to form two HOOH plus 1O2 or 3O2. For a system with 18O H2O, this mechanism leads to a 2.2:1 ratio of 16O:18O in the product HOOH, in good agreement with the ratio 2.2:1 observed in isotope experiments by Wentworth et al. In this paper we use docking and molecular dynamics techniques (HierDock) to search various protein structures for sites that stabilize these products and intermediates predicted from quantum mechanical calculations. We find that the reaction intermediates for production of HOOH from 1O2 are stabilized at the interface of light and heavy chains of antibodies and T cell receptors. This inter Greek key domain interface structure is unique to antibodies and T cell receptors, but is not present in beta2-microglobulin, which does not show any stabilization in our docking studies. This result is consistent with the experimentally observed lack of HOOH production in this system. Our results provide a plausible mechanism for the reactions and provide an explanation of the specific structural character of antibodies responsible for this unexpected chemistry. PMID- 11880617 TI - The role of n-3 polyunsaturated fatty acids in brain: modulation of rat brain gene expression by dietary n-3 fatty acids. AB - Rats were fed either a high linolenic acid (perilla oil) or high eicosapentaenoic + docosahexaenoic acid (fish oil) diet (8%), and the fatty acid and molecular species composition of ethanolamine phosphoglycerides was determined. Gene expression pattern resulting from the feeding of n-3 fatty acids also was studied. Perilla oil feeding, in contrast to fish oil feeding, was not reflected in total fatty acid composition of ethanolamine phosphoglycerides. Levels of the alkenylacyl subclass of ethanolamine phosphoglycerides increased in response to feeding. Similarly, levels of diacyl phosphatidylethanolamine molecular species containing docosahexaenoic acid (18:0/22:6) were higher in perilla-fed or fish oil-fed rat brains whereas those in ethanolamine plasmalogens remained unchanged. Because plasmalogen levels in the brains of rats fed a n-3 fatty acid-enriched diet increased, it is plausible, however, that docosahexaenoic acid taken up from the food or formed from linolenic acid was deposited in this phospholipid subclass. Using cDNA microarrays, 55 genes were found to be overexpressed and 47 were suppressed relative to controls by both dietary regimens. The altered genes included those controlling synaptic plasticity, cytosceleton and membrane association, signal transduction, ion channel formation, energy metabolism, and regulatory proteins. This effect seems to be independent of the chain length of fatty acids, but the n-3 structure appears to be important. Because n-3 polyunsaturated fatty acids have been shown to play an important role in maintaining normal mental functions and docosahexaenoic acid-containing ethanolamine phosphoglyceride (18:0/22:6) molecular species accumulated in response to n-3 fatty acid feeding, a casual relationship between the two events can be surmised. PMID- 11880619 TI - A cofactor approach to copper-dependent catalytic antibodies. AB - A strategy for the preparation of semisynthetic copper(II)-based catalytic metalloproteins is described in which a metal-binding bis-imidazole cofactor is incorporated into the combining site of the aldolase antibody 38C2. Antibody 38C2 features a large hydrophobic-combining site pocket with a highly nucleophilic lysine residue, Lys(H93), that can be covalently modified. A comparison of several lactone and anhydride reagents shows that the latter are the most effective and general derivatizing agents for the 38C2 Lys residue. A bis imidazole anhydride (5) was efficiently prepared from N-methyl imidazole. The 38C2-5-Cu conjugate was prepared by either (i) initial derivatization of 38C2 with 5 followed by metallation with CuCl2, or (ii) precoordination of 5 with CuCl2 followed by conjugation with 38C2. The resulting 38C2-5-Cu conjugate was an active catalyst for the hydrolysis of the coordinating picolinate ester 11, following Michaelis-Menten kinetics [kcat(11) = 2.3 min(-1) and Km(11) 2.2 mM] with a rate enhancement [kcat(11)k(uncat)(11)] of 2.1 x 10(5). Comparison of the second-order rate constants of the modified 38C2 and the Cu(II)-bis-imidazolyl complex k(6-CuCl2) gives a rate enhancement of 3.5 x 10(4) in favor of the antibody complex with an effective molarity of 76.7 M, revealing a significant catalytic benefit to the binding of the bis-imidazolyl ligand into 38C2. PMID- 11880620 TI - Proteome-scale purification of human proteins from bacteria. AB - The completion of the human genome project and the development of high-throughput approaches herald a dramatic acceleration in the pace of biological research. One of the most compelling next steps will be learning the functional roles of all proteins. Achievement of this goal depends in part on the rapid expression and isolation of proteins at large scale. We exploited recombinational cloning to facilitate the development of methods for the high-throughput purification of human proteins. cDNAs were introduced into a master vector from which they could be rapidly transferred into a variety of protein expression vectors for further analysis. A test set of 32 sequence-verified human cDNAs of various sizes and activities was moved into four different expression vectors encoding different affinity-purification tags. By means of an automatable 2-hr protein purification procedure, all 128 proteins were purified and subsequently characterized for yield, purity, and steps at which losses occurred. Under denaturing conditions when the His6 tag was used, 84% of samples were purified. Under nondenaturing conditions, both the glutathione S-transferase and maltose-binding protein tags were successful in 81% of samples. The developed methods were applied to a larger set of 336 randomly selected cDNAs. Sixty percent of these proteins were successfully purified under denaturing conditions and 82% of these under nondenaturing conditions. A relational database, FLEXProt, was built to compare properties of proteins that were successfully purified and proteins that were not. We observed that some domains in the Pfam database were found almost exclusively in proteins that were successfully purified and thus may have predictive character. PMID- 11880622 TI - Transfer RNA-dependent amino acid biosynthesis: an essential route to asparagine formation. AB - Biochemical experiments and genomic sequence analysis showed that Deinococcus radiodurans and Thermus thermophilus do not possess asparagine synthetase (encoded by asnA or asnB), the enzyme forming asparagine from aspartate. Instead these organisms derive asparagine from asparaginyl-tRNA, which is made from aspartate in the tRNA-dependent transamidation pathway [Becker, H. D. & Kern, D. (1998) Proc. Natl. Acad. Sci. USA 95, 12832-12837; and Curnow, A. W., Tumbula, D. L., Pelaschier, J. T., Min, B. & Soll, D. (1998) Proc. Natl. Acad. Sci. USA 95, 12838-12843]. A genetic knockout disrupting this pathway deprives D. radiodurans of the ability to synthesize asparagine and confers asparagine auxotrophy. The organism's capacity to make asparagine could be restored by transformation with Escherichia coli asnB. This result demonstrates that in Deinococcus, the only route to asparagine is via asparaginyl-tRNA. Analysis of the completed genomes of many bacteria reveal that, barring the existence of an unknown pathway of asparagine biosynthesis, a wide spectrum of bacteria rely on the tRNA-dependent transamidation pathway as the sole route to asparagine. PMID- 11880621 TI - Mot1 activates and represses transcription by direct, ATPase-dependent mechanisms. AB - Mot1 is an essential yeast Snf2/Swi2-related ATPase that exerts both positive and negative effects on gene expression. In vitro, Mot1 can disrupt TATA-binding protein-DNA complexes in an ATP-dependent reaction. This activity can explain Mot1-mediated transcriptional repression, but how Mot1 activates transcription is unknown. We demonstrate that, remarkably, Mot1 is localized in vivo to promoters for both Mot1-repressed and Mot1-activated genes. Moreover, Mot1 ATPase activity is required for both activation and repression of gene activity. These findings suggest a novel function for the Mot1 ATPase at activated genes, perhaps involving ATP-driven reorganization of the preinitiation complex. Mot1 regulates the expression of approximately 3% of yeast genes in cells grown in rich medium. Most of these genes are repressed by Mot1, consistent with Mot1's ATP-dependent TATA-binding protein-DNA dissociating activity. Additionally, approximately 77% of the Mot1-repressed genes are involved in the diauxic shift, stress response, mating, or sporulation. The gene sets controlled by NC2 and Srb10 are strongly correlated with the Mot1-controlled set, suggesting that these factors cooperate in transcriptional control on a global scale. PMID- 11880623 TI - Interactions among the protein and RNA subunits of Saccharomyces cerevisiae nuclear RNase P. AB - Ribonuclease P (RNase P) is a ubiquitous endoribonuclease that cleaves precursor tRNAs to generate mature 5' termini. Although RNase P from all kingdoms of life have been found to have essential RNA subunits, the number and size of the protein subunits ranges from one small protein in bacteria to at least nine proteins of up to 100 kDa. In Saccharomyces cerevisiae nuclear RNase P, the enzyme is composed of ten subunits: a single RNA and nine essential proteins. The spatial organization of these components within the enzyme is not yet understood. In this study we examine the likely binary protein-protein and protein-RNA subunit interactions by using directed two- and three-hybrid tests in yeast. Only two protein subunits, Pop1p and Pop4p, specifically bind the RNA subunit. Pop4p also interacted with seven of the other eight protein subunits. The remaining protein subunits all showed one or more specific protein-protein interactions with the other integral protein subunits. Of particular interest was the behavior of Rpr2p, the only protein subunit found in RNase P but not in the closely related enzyme, RNase MRP. Rpr2p interacts strongly with itself as well as with Pop4p. Similar interactions with self and Pop4p were also detected for Snm1p, the only unique protein subunit so far identified in RNase MRP. This observation is consistent with Snm1p and Rpr2p serving analogous functions in the two enzymes. This study provides a low-resolution map of the multisubunit architecture of the ribonucleoprotein enzyme, nuclear RNase P from S. cerevisiae. PMID- 11880624 TI - Microcin E492, a channel-forming bacteriocin from Klebsiella pneumoniae, induces apoptosis in some human cell lines. AB - The cytotoxic effect of microcin E492, a low-molecular-mass channel-forming bacteriocin (7,887 Da) produced by a strain of Klebsiella pneumoniae, was characterized in HeLa cells. At low (5 microg/ml) and intermediate (10 microg/ml) concentrations, microcin E492 induced biochemical and morphological changes typical of apoptosis, such as cell shrinkage, DNA fragmentation, extracellular exposure of phosphatidylserine, caspase activation, and loss of mitochondrial membrane potential. Treatment with zVAD-fmk, a general caspase inhibitor, completely blocked the cytotoxic effect of this bacteriocin. At higher microcin concentrations (>20 microg/ml) a necrotic phenotype was observed. Induction of apoptosis by microcin E492 was associated with the release of calcium from intracellular stores, probably after microcin-triggered ion channel formation. Microcin E492 also presented a cytotoxic effect on Jurkat and RJ2.25 cells, but had no effect on KG-1 cells nor on a primary culture of human tonsil endothelial cells, suggesting that there is a specific interaction of the bacteriocin with components of the target cell surface. This report describes a bacteriocin that has the capacity to induce apoptosis in human cell lines. PMID- 11880625 TI - 13C NMR isotopomer analysis reveals a connection between pyruvate cycling and glucose-stimulated insulin secretion (GSIS). AB - Cellular metabolism of glucose is required for stimulation of insulin secretion from pancreatic beta cells, but the precise metabolic coupling factors involved in this process are not known. In an effort to better understand mechanisms of fuel-mediated insulin secretion, we have adapted 13C NMR and isotopomer methods to measure influx of metabolic fuels into the tricarboxylic acid (TCA) cycle in insulinoma cells. Mitochondrial metabolism of [U-13C3]pyruvate, derived from [U 13C6]glucose, was compared in four clonal rat insulinoma cell 1-derived cell lines with varying degrees of glucose responsiveness. A 13C isotopomer analysis of glutamate isolated from these cells showed that the fraction of acetyl-CoA derived from [U-13C6]glucose was the same in all four cell lines (44 +/- 5%, 70 +/- 3%, and 84 +/- 4% with 3, 6, or 12 mM glucose, respectively). The 13C NMR spectra also demonstrated the existence of two compartmental pools of pyruvate, one that exchanges with TCA cycle intermediates and a second pool derived from [U 13C6]glucose that feeds acetyl-CoA into the TCA cycle. The 13C NMR spectra were consistent with a metabolic model where the two pyruvate pools do not randomly mix. Flux between the mitochondrial intermediates and the first pool of pyruvate (pyruvate cycling) varied in proportion to glucose responsiveness in the four cell lines. Furthermore, stimulation of pyruvate cycling with dimethylmalate or its inhibition with phenylacetic acid led to proportional changes in insulin secretion. These findings indicate that exchange of pyruvate with TCA cycle intermediates, rather than oxidation of pyruvate via acetyl-CoA, correlates with glucose-stimulated insulin secretion. PMID- 11880626 TI - Nucleotides and phospholipids compete for binding to the C terminus of KATP channels. AB - Inwardly rectifying, ATP-sensitive K+ channels (K(ATP)) couple metabolism to either cell excitability (Kir6.x) or potassium secretion (Kir1.1). Phosphatidylinositol phospholipids, like PI(4,5)P2, antagonize nucleotide inhibition of K(ATP) channels enhancing the coupling of metabolic events to cell electrical or transport activity. The mechanism by which phospholipids relieve ATP block is unclear. We have shown that maltose-binding fusion proteins (MBP) containing the COOH termini of K(ATP) channels (Kir1.1, Kir6.1, and Kir6.2) form functional tetramers that directly bind at least two ATP molecules with negative cooperativity. Here we show that purified phosphatidylinositol phospholipids compete for 2,4,6,-trinitrophenyl (TNP)-ATP binding to the COOH termini of K(ATP) channels with EC50 values for PIP2 between 6-8 microM. The phospholipid potency profile was PIP3 > PIP2 = PIP > PI, suggesting that net phospholipid charge was important. A role for head group charge was supported by polycations (neomycin, spermine, and polylysine) reversing the effect of PIP2 on TNP-ATP binding to the Kir1.1 channel COOH terminal fusion protein. In contrast, the water-soluble charged hydrolytic product of PIP2, inositol(1,4,5)P3 (IP3), had no effect on TNP ATP binding, suggesting that the acyl chain of PIP2 was also necessary for its effect on TNP-ATP binding. Indeed, neutral and charged lipids had weak, but significant, effects on TNP-ATP binding. Whereas microM concentrations of PIP2 could compete with TNP-ATP, we found that mM concentrations of MgATP were required to compete with PIP2 for binding to these K(ATP) channel COOH termini. Thus the COOH termini of K(ATP) channels form a nucleotide- and phospholipid modulated channel gate on which ATP and phospholipids compete for binding. PMID- 11880627 TI - Natural beta-sheet proteins use negative design to avoid edge-to-edge aggregation. AB - The fact that natural beta-sheet proteins are usually soluble but that fragments or designs of beta structure usually aggregate suggests that natural beta proteins must somehow be designed to avoid this problem. Regular beta-sheet edges are dangerous, because they are already in the right conformation to interact with any other beta strand they encounter. We surveyed edge strands in a large sample of all-beta proteins to tabulate features that could protect against further beta-sheet interactions. beta-barrels, of course, avoid edges altogether by continuous H-bonding around the barrel cylinder. Parallel beta-helix proteins protect their beta-sheet ends by covering them with loops of other structure. beta-propeller and single-sheet proteins use a combination of beta-bulges, prolines, strategically placed charges, very short edge strands, and loop coverage. beta-sandwich proteins favor placing an inward-pointing charged side chain on one of the edge strands where it would be buried by dimerization; they also use bulges, prolines, and other mechanisms. One recent beta-hairpin design has a constrained twist too great for accommodation into a larger beta-sheet, whereas some beta-sheet edges are protected by the bend and reverse twist produced by an Lbeta glycine. All free edge strands were seen to be protected, usually by several redundant mechanisms. In contrast, edge strands that natively form beta H-bonded dimers or rings have long, regular stretches without such protection. These results are relevant to understanding how proteins may assemble into beta-sheet amyloid fibers, and they are especially applicable to the de novo design of beta structure. Many edge-protection strategies used by natural proteins are beyond our current abilities to constrain by design, but one possibility stands out as especially useful: a single charged side chain near the middle of what would ordinarily be the hydrophobic side of the edge beta strand. This minimal negative-design strategy changes only one residue, requires no backbone distortion, and is easy to design. The accompanying paper [Wang, W. & Hecht, M. H. (2002) Proc. Natl. Acad. Sci. USA 99, 2760-2765] makes use of the inward-pointing charge strategy with great success, turning highly aggregated beta-sandwich designs into soluble monomers. PMID- 11880628 TI - Rationally designed mutations convert de novo amyloid-like fibrils into monomeric beta-sheet proteins. AB - Amyloid fibrils are associated with a variety of neurodegenerative maladies including Alzheimer's disease and the prion diseases. The structures of amyloid fibrils are composed of beta-strands oriented orthogonal to the fibril axis ("cross beta" structure). We previously reported the design and characterization of a combinatorial library of de novo beta-sheet proteins that self-assemble into fibrillar structures resembling amyloid. The libraries were designed by using a "binary code" strategy, in which the locations of polar and nonpolar residues are specified explicitly, but the identities of these residues are not specified and are varied combinatorially. The initial libraries were designed to encode proteins containing amphiphilic beta-strands separated by reverse turns. Each beta-strand was designed to be seven residues long, with polar (open circle) and nonpolar (shaded circle) amino acids arranged with an alternating periodicity ([see text]). The initial design specified the identical polar/nonpolar pattern for all of the beta-strands; no strand was explicitly designated to form the edges of the resulting beta-sheets. With all beta-strands preferring to occupy interior (as opposed to edge) locations, intermolecular oligomerization was favored, and the proteins assembled into amyloid-like fibrils. To assess whether explicit design of edge-favoring strands might tip the balance in favor of monomeric beta-sheet proteins, we have now redesigned the first and/or last beta strands of several sequences from the original library. In the redesigned beta strands, the binary pattern is changed from [see text] (K denotes lysine). The presence of a lysine on the nonpolar face of a beta-strand should disfavor fibrillar structures because such structures would bury an uncompensated charge. The nonpolar right arrow lysine mutations, therefore, would be expected to favor monomeric structures in which the [see text] sequences form edge strands with the charged lysine side chain accessible to solvent. To test this hypothesis, we constructed several second generation sequences in which the central nonpolar residue of either the N-terminal beta-strand or the C-terminal beta-strand (or both) is changed to lysine. Characterization of the redesigned proteins shows that they form monomeric beta-sheet proteins. PMID- 11880629 TI - Two-photon excitation microscopy of tryptophan-containing proteins. AB - We have examined the feasibility of observing single protein molecules by means of their intrinsic tryptophan emission after two-photon excitation. A respiratory protein from spiders, the 24-meric hemocyanin, containing 148 tryptophans, was studied in its native state under almost in vivo conditions. In this specific case, the intensity of the tryptophan emission signals the oxygen load, allowing one to investigate molecular cooperativity. As a system with even higher tryptophan content, we also investigated latex spheres covered with the protein avidin, resulting in 340 tryptophans per sphere. The ratio of the fluorescence quantum efficiency to the bleaching efficiency was found to vary between 2 and 180 after two-photon excitation for tryptophan free in buffer solution, in hemocyanin, and in avidin-coated spheres. In the case of hemocyanin, this ratio leads to about four photons detected before photobleaching. Although this number is quite small, the diffusion of individual protein molecules could be detected by fluorescence correlation spectroscopy. In avidin-coated spheres, the tryptophans exhibit a higher photostability, so that even imaging of single spheres becomes possible. As an unexpected result of the measurements, it was discovered that the population of the oxygenated state of hemocyanin can be changed by means of a one-photon process with the same laser source that monitors this population in a two-photon process. PMID- 11880630 TI - Proton shuttle in green fluorescent protein studied by dynamic simulations. AB - As a direct simulation of a multistep proton transfer reaction involving protein residues, the proton relay shuttle between A and I forms of green fluorescent protein (GFP) is simulated in atomic detail by using a special molecular dynamics simulation technique. Electronic excitation of neutral chromophore in wild-type GFP is generally followed by excited-state proton transfer to a nearby glutamic acid residue via a water molecule and a serine residue. Here we show that the second and third transfer steps occur ultrafast on time scales of several tens of femtoseconds. Proton back-shuttle in the ground state is slower and occurs in a different sequence of events. The simulations provide atomic models of various intermediates and yield realistic rate constants for proton transfer events. In particular, we argue that the I form observed spectroscopically under equilibrium conditions may differ from the I form observed as a fast intermediate by an anti to syn rotation of the carboxyl proton of neutral Glu-222. PMID- 11880632 TI - Crystal structure of an antiparallel DNA fragment with Hoogsteen base pairing. AB - We report here an alternative double-helical structure of the DNA molecule. It has been found in the d(ATA(Br)UAT) and d(ATATAT) sequences by single-crystal x ray crystallography. This sequence is found not only in TATA boxes, but also in other regulatory regions of DNA. Bases of the two antiparallel strands form Hoogsteen pairs, with adenines in the syn conformation. The structure is related neither to those found in triple helices nor to parallel DNA duplexes. Its conformational parameters are very similar to those of duplex DNA in the B form. Both forms may coexist under physiological conditions, although the Hoogsteen pairing greatly influences the recognition sites on DNA. Our results demonstrate that an alternative to the classical B-DNA double helix is possible. PMID- 11880631 TI - Crystal structure of the yeast cytochrome bc1 complex with its bound substrate cytochrome c. AB - Small diffusible redox proteins facilitate electron transfer in respiration and photosynthesis by alternately binding to integral membrane proteins. Specific and transient complexes need to be formed between the redox partners to ensure fast turnover. In respiration, the mobile electron carrier cytochrome c shuttles electrons from the cytochrome bc1 complex to cytochrome c oxidase. Despite extensive studies of this fundamental step of energy metabolism, the structures of the respective electron transfer complexes were not known. Here we present the crystal structure of the complex between cytochrome c and the cytochrome bc1 complex from Saccharomyces cerevisiae. The complex was crystallized with the help of an antibody fragment, and its structure was determined at 2.97-A resolution. Cytochrome c is bound to subunit cytochrome c1 of the enzyme. The tight and specific interactions critical for electron transfer are mediated mainly by nonpolar forces. The close spatial arrangement of the c-type hemes unexpectedly suggests a direct and rapid heme-to-heme electron transfer at a calculated rate of up to 8.3 x 10(6) s(-1). Remarkably, cytochrome c binds to only one recognition site of the homodimeric multisubunit complex. Interestingly, the occupancy of quinone in the Qi site is higher in the monomer with bound cytochrome c, suggesting a coordinated binding and reduction of both electron accepting substrates. Obviously, cytochrome c reduction by the cytochrome bc1 complex can be regulated in response to respiratory conditions. PMID- 11880633 TI - Polarized cytokinesis in vacuolate cells of Arabidopsis. AB - The view of plant-cell cytokinesis commonly depicted in textbooks is of a symmetrical process, with the phragmoplast initiating in the center of the cell and growing outward to the parental cell membrane. In contrast to this picture, we observe that cell-plate development in Arabidopsis shoot cells is highly polarized along the plane of division. Three-dimensional live-cell imaging reveals that the mitotic spindle and phragmoplast are laterally displaced, and that the growing cell plate anchors on one side of the cell at an early stage of cytokinesis. Growth of phragmoplast across the cell creates a new partition in its wake, giving the visual effect of a curtain being pulled across the cell. Throughout this process, the advancing front of the phragmoplast is in intimate contact with the parental wall, suggesting that short-range interactions between the phragmoplast and plasma membrane may play important roles in guiding the cell plate throughout much of its development. Polarized cytokinesis was observed in a wide variety of vacuolate shoot cells and in some small root cells, implying that it is not solely a function of cell size. This mode of cytokinesis may provide a mechanically robust mechanism for cell-plate formation in large cells and suggests a simple explanation for the occurrence of cell wall stubs observed upon drug treatment or in cytokinetic mutants. PMID- 11880634 TI - Phosphatidylinositol-dependent actin filament binding by the SWI/SNF-like BAF chromatin remodeling complex. AB - Recently, several chromatin remodeling complexes in yeast, Drosophila, and mammals have been shown to contain actin and actin-related proteins (arps). However, the function of actin in these complexes is unclear. Here, we show that the mammalian SWI/SNF-like BAF complex binds to phosphatidylinositol 4,5 bisphosphate (PIP2) micelles and PIP2-containing mixed lipid vesicles, and that PIP2 binding allows the complex to associate with actin pointed ends and branch points. Actin binds to at least two distinct domains in the C terminus of the Brg1 protein, and interaction with only one of these domains is sensitive to PIP2. Based on these findings, we propose a model for PIP2 activation of actin binding by relief of intramolecular capping of actin by Brg1. PMID- 11880635 TI - Expression profiling of a human cell line model of prostatic cancer reveals a direct involvement of interferon signaling in prostate tumor progression. AB - Cancer-associated fibroblasts induce malignant behavior in genetically initiated but nontumorigenic human prostatic epithelium. The genetic basis for such transformation is still unknown. By using Affymetrix GeneChip technology, we profiled genomewide gene expression of transformed [tumorigenic benign prostatic hyperplasia (BPH1)(CAFTD)] and parental (nontumorigenic BPH1) cells. We identified differentially expressed genes, which are associated with tumorigenesis or tumor progression. One striking finding is that a significant portion of the down-regulated genes belongs to interferon (IFN)-inducible molecules. We show that IFN inhibited the tumorigenic BPH1(CAFTD) cell proliferation and colony formation in vitro and inhibited tumor growth in xenografts in vivo. Expression of the IFN-inducible molecules correlates with the growth-inhibiting effects of IFN. In addition, these genes are reported to be mapped mainly to two chromosomal regions, 10q23-26 and 17q21, which are frequently deleted in human prostate cancers. Furthermore, in silico data-mining with the GeneLogic database revealed that expression of the IFN-inducible genes was down-regulated in approximately 30% of the 49 clinically characterized samples of prostatic adenocarcinomas. Collectively, we show that there seems to be a direct link between IFN-inducible molecules and prostatic tumor progression. These findings suggest IFN-inducible molecules as potential therapeutic targets for the treatment of prostate cancer. PMID- 11880637 TI - Binding of rainbow trout sperm to egg is mediated by strong carbohydrate-to carbohydrate interaction between (KDN)GM3 (deaminated neuraminyl ganglioside) and Gg3-like epitope. AB - KDNalpha2-->3Galbeta4Glcbeta1Cer [(KDN)GM3] is a major (approximately 90%) component of total gangliosides found in sperm of rainbow trout (Oncorhynchus mykiss) and was shown to be present prominently at the sperm head by immunochemical staining with its specific mAb kdn3G. Liposomes containing (KDN)GM3 adhere specifically to GalNAcbeta4Galbeta4Glcbeta1Cer (Gg3Cer)-coated plastic plates. Interaction between (KDN)GM3 and Gg3Cer was much stronger than that previously observed between Neu5Acalpha2-->3Galbeta4Glcbeta1Cer and Gg3Cer. (KDN)GM3-Gg3Cer interaction did not require the presence of Ca2+ and Mg2+, but was enhanced in the presence of Mn2+. Fresh trout sperm adhered specifically to Gg3Cer-coated plates under physiological conditions, and the binding was inhibited by pretreatment of sperm with mAb kdn3G. The presence of Gg3 or Gg3 related epitope structure in the specific area surrounding the micropyle, through which sperm enter the egg, was confirmed by immunogold labeling under electron microscopy. These findings suggest that initial sperm-egg adhesion during the process of fertilization occurs when sperm adhere to the area surrounding the micropyle through specific interaction between (KDN)GM3 on the sperm head and Gg3 epitope (GalNAcbeta4Galbeta1-->) expressed at a defined region of the egg surface membrane. PMID- 11880636 TI - DEC1 is a downstream target of TGF-beta with sequence-specific transcriptional repressor activities. AB - To identify genes that mediate transforming growth factor-beta (TGF-beta) signaling, a colorectal cancer cell line that was sensitive to the growth inhibitory effects of this cytokine was created. We then determined the global gene expression profiles of these cells, and those of HaCaT human keratinocytes, in the presence and absence of TGF-beta. Of the several genes identified in this screen, DEC1 was of particular note in light of the rapidity and consistency of its induction and its potential biochemical activities. We identified a consensus DNA-binding site for DEC1 and showed that DEC1 could repress the transcription of a reporter containing this binding site in its promoter. Finally, both alleles of the DEC1 locus in HaCaT cells were inactivated through targeted homologous recombination. This approach revealed that DEC1 induction was not required for the growth inhibition mediated by TGF-beta in this line. However, DEC1 may function in concert with other signaling components to mediate certain biologic effects of TGF-beta. PMID- 11880638 TI - Inference of functional regions in proteins by quantification of evolutionary constraints. AB - Likelihood estimates of local rates of evolution within proteins reveal that selective constraints on structure and function are quantitatively stable over billions of years of divergence. The stability of constraints produces an intramolecular clock that gives each protein a characteristic pattern of evolutionary rates along its sequence. This pattern allows the identification of constrained regions and, because the rate of evolution is a quantitative measure of the strength of the constraint, of their functional importance. We show that results from such analyses, which require only sequence alignments, are consistent with experimental and mutational data. The methodology has significant predictive power and may be used to guide structure--function studies for any protein represented by a modest number of homologs in sequence databases. PMID- 11880639 TI - Wolbachia density and virulence attenuation after transfer into a novel host. AB - The factors that control replication rate of the intracellular bacterium Wolbachia pipientis in its insect hosts are unknown and difficult to explore, given the complex interaction of symbiont and host genotypes. Using a strain of Wolbachia that is known to over-replicate and shorten the lifespan of its Drosophila melanogaster host, we have tracked the evolution of replication control in both somatic and reproductive tissues in a novel host/Wolbachia association. After transinfection (the transfer of a Wolbachia strain into a different species) of the over-replicating Wolbachia popcorn strain from D. melanogaster to Drosophila simulans, we demonstrated that initial high densities in the ovaries were in excess of what was required for perfect maternal transmission, and were likely causing reductions in reproductive fitness. Both densities and fitness costs associated with ovary infection rapidly declined in the generations after transinfection. The early death effect in D. simulans attenuated only slightly and was comparable to that induced in D. melanogaster. This study reveals a strong host involvement in Wolbachia replication rates, the independence of density control responses in different tissues, and the strength of natural selection acting on reproductive fitness. PMID- 11880640 TI - Genome complexity reduction for SNP genotyping analysis. AB - Efficient single nucleotide polymorphism (SNP) genotyping methods are necessary to accomplish many current gene discovery goals. A crucial element in large-scale SNP genotyping is the number of individual biochemical reactions that must be performed. An efficient method that can be used to simultaneously amplify a set of genetic loci across a genome with high reliability can provide a valuable tool for large-scale SNP genotyping studies. In this paper we describe and characterize a method that addresses this goal. We have developed a strategy for reducing genome complexity by using degenerate oligonucleotide primer (DOP)-PCR and applied this strategy to SNP genotyping in three complex eukaryotic genomes; human, mouse, and Arabidopsis thaliana. Using a single DOP-PCR primer, SNP loci spread throughout a genome can be amplified and accurately genotyped directly from a DOP-PCR product mixture. DOP-PCRs are extremely reproducible. The DOP-PCR method is transferable to many species of interest. Finally, we describe an in silico approach that can effectively predict the SNP loci amplified in a given DOP-PCR, permitting the design of an efficient set of reactions for large-scale, genome-wide SNP studies. PMID- 11880641 TI - The organization of Physcomitrella patensRAD51 genes is unique among eukaryotic organisms. AB - Genetic recombination pathways and genes are well studied, but relatively little is known in plants, especially in lower plants. To study the recombination apparatus of a lower land plant, a recombination gene well characterized particularly in yeast, mouse, and man, the RAD51 gene, was isolated from the moss Physcomitrella patens and characterized. Two highly homologous RAD51 genes were found to be present. Duplicated RAD51 genes have been found thus far exclusively in eukaryotes with duplicated genomes. Therefore the presence of two highly homologous genes suggests a recent genome duplication event in the ancestry of Physcomitrella. Comparison of the protein sequences to Rad51 proteins from other organisms showed that both RAD51 genes originated within the group of plant Rad51 proteins. However, the two proteins form a separate clade in a phylogenetic tree of plant Rad51 proteins. In contrast to RAD51 genes from other multicellular eukaryotes, the Physcomitrella genes are not interrupted by introns. Because introns are a common feature of Physcomitrella genes, the lack of introns in the RAD51 genes is unusual and may indicate the presence of an unusual recombination apparatus in this organism. The presence of duplicated intronless RAD51 genes is unique among eukaryotes. Studies of further members of this lineage are needed to determine whether this feature may be typical of lower plants. PMID- 11880642 TI - Different contributions of thymopoiesis and homeostasis-driven proliferation to the reconstitution of naive and memory T cell compartments. AB - Following transfer into lymphopenic hosts, naive CD8 T cells proliferate and acquire memory phenotype. Although the acquired phenotype is stable in recombination activating gene-1-deficient (RAG-/-) recipients, in sublethally irradiated mice naive CD8 T cells of donor origin gradually accumulate. The naive cells have been attributed to phenotypic reversion of homeostatic memory cells, implying instability of memory phenotype and restoration of the naive T cell compartment by homeostasis-driven proliferation. We show here that (i) the accumulation of naive CD8 T cells of donor origin only occurs in recipients that have been irradiated and have an intact thymus; (ii) the apparent reversion of memory to naive cells actually results from de novo T cell development of hematopoietic stem cells, present in the donor spleen or lymph node cell populations, in the thymus of irradiated recipients; and (iii) the number of homeostatic memory cells generated in both RAG-/- and irradiated hosts reaches a plateau value and their phenotype is stably maintained even after retransfer into nonirradiated normal mice for 30 days. These findings demonstrate that homeostatic memory T cells do not revert to naive cells. After severe T cell depletion homeostasis-driven proliferation restores only the memory T cell compartment, whereas thymopoiesis is required for the reconstitution of the naive T cell compartment. PMID- 11880643 TI - West Nile virus/dengue type 4 virus chimeras that are reduced in neurovirulence and peripheral virulence without loss of immunogenicity or protective efficacy. AB - A candidate live attenuated vaccine strain was constructed for West Nile virus (WN), a neurotropic flavivirus that has recently emerged in the U.S. Considerable attenuation for mice was achieved by chimerization with dengue virus type 4 (DEN4). The genes for the structural premembrane and envelope proteins of DEN4 present in an infectious cDNA clone were replaced by the corresponding genes of WN strain NY99. Two of 18 cDNA clones of a WN/DEN4 chimera yielded full-length RNA transcripts that were infectious when transfected into susceptible cells. The two infectious clones shared a motif in the transmembrane signal domain located immediately downstream of the NS2B-NS3 protease cleavage site that separates the DEN4 capsid protein and the WN premembrane protein of the chimera. This motif, Asp and Thr at a position 3 and 6 amino acids downstream of the cleavage site, respectively, was not present in the 16 noninfectious cDNA clones. The WN/DEN4 chimera was highly attenuated in mice compared with its WN parent; the chimera was at least 28,500 times less neurovirulent in suckling mice inoculated intracerebrally and at least 10,000 times less virulent in adult mice inoculated intraperitoneally. Nonetheless, the WN/DEN4 chimera and a deletion mutant derived from it were immunogenic and provided complete protection against lethal WN challenge. These observations provide the basis for pursuing the development of a live attenuated WN vaccine. PMID- 11880644 TI - Primary role for adherent leukocytes in sickle cell vascular occlusion: a new paradigm. AB - Vascular occlusion is the major cause of morbidity and mortality in sickle cell disease but its mechanisms are poorly understood. We demonstrate by using intravital microscopy in mice expressing human sickle hemoglobin (SS) that SS red blood cells (RBCs) bind to adherent leukocytes in inflamed venules, producing vasoocclusion of cremasteric venules. SS mice deficient in P- and E-selectins, which display defective leukocyte recruitment to the vessel wall, are protected from vasoocclusion. These data uncover a previously unsuspected paradigm for the pathogenesis of sickle cell vasoocclusion in which adherent leukocytes play a direct role and suggest that drugs targeting SS RBC-leukocyte or leukocyte endothelial interactions may prevent or treat the vascular complications of this debilitating disease. PMID- 11880645 TI - PATE, a gene expressed in prostate cancer, normal prostate, and testis, identified by a functional genomic approach. AB - To identify target antigens for prostate cancer therapy, we have combined computer-based screening of the human expressed sequence tag database and experimental expression analysis to identify genes that are expressed in normal prostate and prostate cancer but not in essential human tissues. Using this approach, we identified a gene that is expressed specifically in prostate cancer, normal prostate, and testis. The gene has a 1.5-kb transcript that encodes a protein of 14 kDa. We named this gene PATE (expressed in prostate and testis). In situ hybridization shows that PATE mRNA is expressed in the epithelial cells of prostate cancers and in normal prostate. Transfection of the PATE cDNA with a Myc epitope tag into NIH 3T3 cells and subsequent cell fractionation analysis shows that the PATE protein is localized in the membrane fraction of the cell. Analysis of the amino acid sequence of PATE shows that it has structural similarities to a group of proteins known as three-finger toxins, which includes the extracellular domain of the type beta transforming growth factor receptor. Restricted expression of PATE makes it a potential candidate for the immunotherapy of prostate cancer. PMID- 11880646 TI - A prototypic platelet septin and its participation in secretion. AB - Studies are presented characterizing platelet CDCrel-1, a protein expressed to high levels by megakaryocytes and belonging to a family of conserved proteins, termed septin. Septin filaments originally were identified in yeast as essential for budding but have become increasingly associated with processes in higher eukaryotic cells involving active membrane movement such as cytokinesis and vesicle trafficking. Direct proof of an in vivo function for septins in higher eukaryotes is limited to the characterization of the Drosophila septin, termed PNUT. We present studies identifying platelet CDCrel-1 as a protein kinase substrate in the presence of known platelet agonists. The immunopurification of CDCrel-1 revealed it to be part of a macromolecular complex containing a protein involved in platelet secretion, syntaxin 4. Moreover, CDCrel-1 was localized in situ to areas surrounding platelet-storage granules. The relevance of CDCrel-1 to normal platelet function was established with the characterization of platelets from a CDCrel-1(Null) mouse. As compared with platelets from wild-type littermates, CDCrel-1(Null) platelets aggregate and release stored [14C]serotonin in the presence of subthreshold levels of collagen. These results provide new insights into the mechanisms regulating platelet secretion and identify platelet septins as a protein family contributing to membrane trafficking within the megakaryocyte and platelet. PMID- 11880647 TI - Early changes in hepatitis C viral quasispecies during interferon therapy predict the therapeutic outcome. AB - Despite recent treatment advances, the majority of patients with chronic hepatitis C fail to respond to antiviral therapy. Although the genetic basis for this resistance is unknown, accumulated evidence suggests that changes in the heterogeneous viral population (quasispecies) may be an important determinant of viral persistence and response to therapy. Sequences within hepatitis C virus (HCV) envelope 1 and envelope 2 genes, inclusive of the hypervariable region 1, were analyzed in parallel with the level of viral replication in serial serum samples obtained from 23 patients who exhibited different patterns of response to therapy and from untreated controls. Our study provides evidence that although the viral diversity before treatment does not predict the response to treatment, the early emergence and dominance of a single viral variant distinguishes patients who will have a sustained therapeutic response from those who subsequently will experience a breakthrough or relapse. A dramatic reduction in genetic diversity leading to an increasingly homogeneous viral population was a consistent feature associated with viral clearance in sustained responders and was independent of HCV genotype. The persistence of variants present before treatment in patients who fail to respond or who experience a breakthrough during therapy strongly suggests the preexistence of viral strains with inherent resistance to IFN. Thus, the study of the evolution of the HCV quasispecies provides prognostic information as early as the first 2 weeks after starting therapy and opens perspectives for elucidating the mechanisms of treatment failure in chronic hepatitis C. PMID- 11880648 TI - Mycobacterium tuberculosis WhiB3 interacts with RpoV to affect host survival but is dispensable for in vivo growth. AB - Previous work established that the principal sigma factor (RpoV) of virulent Mycobacterium bovis, a member of the Mycobacterium tuberculosis complex, restores virulence to an attenuated strain containing a point mutation (Arg-515-->His) in the 4.2 domain of RpoV. We used the 4.2 domain of RpoV as bait in a yeast two hybrid screen of an M. tuberculosis H37Rv library and identified a putative transcription factor, WhiB3, which selectively interacts with the 4.2 domain of RpoV in virulent strains but not with the mutated (Arg-515-->His) allele. Infection of mice and guinea pigs with a M. tuberculosis H37Rv whiB3 deletion mutant strain showed that whiB3 is not necessary for in vivo bacterial replication in either animal model. In contrast, an M. bovis whiB3 deletion mutant was completely attenuated for growth in guinea pigs. However, we found that immunocompetent mice infected with the M. tuberculosis H37Rv whiB3 mutant strain had significantly longer mean survival times as compared with mice challenged with wild-type M. tuberculosis. Remarkably, the bacterial organ burdens of both mutant and wild-type infected mice were identical during the acute and persistent phases of infection. Our results imply that M. tuberculosis replication per se is not a sufficient condition for virulence in vivo. They also indicate a different role for M. bovis and M. tuberculosis whiB3 genes in pathogenesis generated in different animal models. We propose that M. tuberculosis WhiB3 functions as a transcription factor regulating genes that influence the immune response of the host. PMID- 11880649 TI - Antivirally active MxA protein sequesters La Crosse virus nucleocapsid protein into perinuclear complexes. AB - Bunyaviruses replicate in the cytoplasm of infected cells. New viral particles are formed by budding of nucleocapsids into the Golgi apparatus. We have previously shown that the IFN-induced human MxA protein inhibits bunyavirus replication by an unknown mechanism. Here we demonstrate that MxA binds to the nucleocapsid protein of La Crosse virus (LACV) and colocalizes with the viral protein in cytoplasmic complexes. Electron microscopy revealed that these complexes accumulated in the perinuclear area and consisted of highly ordered fibrillary structures. A similar MxA-mediated redistribution of viral nucleocapsid proteins was detected with other bunyaviruses, such as Bunyamwera virus and Rift Valley fever virus. MxA(E645R), a carboxy-terminal mutant of MxA without antiviral activity against LACV, did not lead to complex formation. Wild type MxA, but not MxA(E645R), was able to bind to LACV nucleocapsid protein in coimmunoprecipitation assays, demonstrating the importance of the carboxy terminal effector domain of MxA. These results illustrate an efficient mechanism of IFN action whereby an essential virus component is trapped in cytoplasmic inclusions and becomes unavailable for the generation of new virus particles. PMID- 11880650 TI - Replication of Cryptococcus neoformans in macrophages is accompanied by phagosomal permeabilization and accumulation of vesicles containing polysaccharide in the cytoplasm. AB - Cryptococcus neoformans (CN), an encapsulated, ubiquitous environmental yeast, is pathogenic for humans, primarily those with compromised immune function. CN is believed to be a facultative intracellular pathogen. Time-lapsed video microscopy revealed that yeast began to replicate and divide 2 hours after ingestion by J774.16 macrophage cells, with the average cell hosting 10-40 organisms of varying morphologies before ultimately lysing and releasing organisms, either singly or in clumps. Intracellular growth was accompanied by the accumulation of polysaccharide-filled vesicles in the macrophage. Studies with fluorescently labeled dextran revealed that the phagolysosomal compartment became leaky during the course of intracellular infection. Consistent with this observation, phagosomes containing CN had an increased pH relative to similar phagosomes containing inert magnetic beads, as indicated by a colorimetric change in the pH sensitive Lysosensor dye. Immunocytochemistry revealed differences in the reactivity of polysaccharide elaborated by CN inside macrophages relative to that expressed in vitro. Taken together these results are suggestive of a novel mechanism of intracellular survival by an encapsulated organism, whereby ingestion is followed by damage to the phagosomal membrane resulting in continuity with the cytoplasm, accumulation of polysaccharide-containing vesicles, and possibly, production of a structurally different polysaccharide. PMID- 11880652 TI - DARPP-32 mediates serotonergic neurotransmission in the forebrain. AB - Serotonin is implicated in the regulation of complex sensory, motor, affective, and cognitive functions. However, the biochemical mechanisms whereby this neurotransmitter exerts its actions remain largely unknown. DARPP-32 (dopamine- and cAMP-regulated phosphoprotein of molecular weight 32,000) is a phosphoprotein that has primarily been characterized in relation to dopaminergic neurotransmission. Here we report that serotonin regulates DARPP-32 phosphorylation both in vitro and in vivo. Stimulation of 5-hydroxy-tryptamine (5 HT4 and 5-HT6 receptors causes an increased phosphorylation state at Thr34-DARPP 32, the protein kinase A site, and a decreased phosphorylation state at Thr75 DARPP-32, the cyclin-dependent kinase 5 site. Furthermore, stimulation of 5-HT2 receptors increases the phosphorylation state of Ser137-DARPP-32, the casein kinase-1 site. Behavioral and gene transcriptional effects induced by compounds that selectively release serotonin were greatly reduced in DARPP-32 knockout mice. Our data indicate that DARPP-32 is essential not only for dopaminergic but also for serotonergic neurotransmission. PMID- 11880653 TI - Order-sensitive plasticity in adult primary auditory cortex. AB - The neural response to a stimulus presented as part of a rapid sequence is often quite different from the response to the same stimulus presented in isolation. In primary auditory cortex (A1), although the most common effect of preceding stimuli is inhibitory, most neurons can also exhibit response facilitation if the appropriate spectral and temporal separation of sequence elements is presented. In this study, we investigated whether A1 neurons in adult animals can develop context-dependent facilitation to a novel acoustic sequence. After repeatedly pairing electrical stimulation of the basal forebrain with a three-element sequence (high frequency tone--low frequency tone-- noise burst), 25% of A1 neurons exhibited facilitation to the low tone when preceded by the high tone, compared with only 5% in controls. In contrast, there was no increase in the percent of sites that showed facilitation for the reversed tone order (low preceding high). Nearly 60% of sites exhibited a facilitated response to the noise burst when preceded by the two tones. Although facilitation was greatest in response to the paired sequence, facilitation also generalized to related sequences that were either temporally distorted or missing one of the tones. Pairing basal forebrain stimulation with the acoustic sequence also caused a decrease in the time to peak response and an increase in population discharge synchrony, which was not seen after pairing simple tones, tone trains, or broadband stimuli. These results indicate that context-dependent facilitation and response synchronization can be substantially altered in an experience-dependent fashion and provide a potential mechanism for learning spectrotemporal patterns. PMID- 11880651 TI - Involvement of striatal and extrastriatal DARPP-32 in biochemical and behavioral effects of fluoxetine (Prozac). AB - Fluoxetine (Prozac) is the most widely prescribed medication for the treatment of depression. Nevertheless, little is known about the molecular basis of its clinical efficacy, apart from the fact that fluoxetine increases the synaptic availability of serotonin. Here we show that, in vivo, fluoxetine, given either acutely or chronically, regulates the phosphorylation state of dopamine- and cAMP regulated phosphoprotein of M(r) 32,000 (DARPP-32) at multiple sites in prefrontal cortex, hippocampus, and striatum. Acute administration of fluoxetine increases phosphorylation of DARPP-32 at the protein kinase A site, Thr-34, and at the casein kinase-1 site, Ser-137, and decreases phosphorylation at the cyclin dependent kinase 5 site, Thr-75. Each of these changes contributes, through distinct signaling pathways, to increased inhibition of protein phosphatase-1, a major serine/threonine protein phosphatase in the brain. Fluoxetine also increases phosphorylation of the alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionic acid (AMPA) receptor subunit GluR1 at Ser-831 and Ser-845. Both the fluoxetine-mediated increase in AMPA receptor phosphorylation at Ser-845 GluR1 and the beneficial responsiveness to fluoxetine in an animal test of antidepressant efficacy were strongly reduced in DARPP-32 knockout mice, indicating a critical role for this phosphoprotein in the antidepressant actions of fluoxetine. Mice chronically treated with fluoxetine had increased levels of DARPP-32 mRNA and protein and a decreased ability to increase phospho-Ser-137 DARPP-32 and phospho-Ser-831-GluR1. These chronic changes may be relevant to the delayed onset of therapeutic efficacy of fluoxetine. PMID- 11880654 TI - Neuronal activity-dependent membrane traffic at the neuromuscular junction. AB - During development and also in adulthood, synaptic connections are modulated by neuronal activity. To follow such modifications in vivo, new genetic tools are designed. The nontoxic C-terminal fragment of tetanus toxin (TTC) fused to a reporter gene such as LacZ retains the retrograde and transsynaptic transport abilities of the holotoxin itself. In this work, the hybrid protein is injected intramuscularly to analyze in vivo the mechanisms of intracellular and transneuronal traffics at the neuromuscular junction (NMJ). Traffic on both sides of the synapse are strongly dependent on presynaptic neural cell activity. In muscle, a directional membrane traffic concentrates beta-galactosidase-TTC hybrid protein into the NMJ postsynaptic side. In neurons, the probe is sorted across the cell to dendrites and subsequently to an interconnected neuron. Such fusion protein, sensitive to presynaptic neuronal activity, would be extremely useful to analyze morphological changes and plasticity at the NMJ. PMID- 11880655 TI - Ligand binding to somatostatin receptors induces receptor-specific oligomer formation in live cells. AB - Heptahelical receptors (HHRs) are generally thought to function as monomeric entities. Several HHRs such as somatostatin receptors (SSTRs), however, form homo and heterooligomers when activated by ligand binding. By using dual fluorescent ligands simultaneously applied to live cells monotransfected with SSTR5 (R5) or SSTR1 (R1), or cotransfected with R5 and R1, we have analyzed the ligand receptor stoichiometry and aggregation states for the three receptor systems by fluorescence resonance energy transfer and fluorescence correlation spectroscopy. Both homo- and heterooligomeric receptors are occupied by two ligand molecules. We find that monomeric, homooligomeric, and heterooligomeric receptor species occur in the same cell cotransfected with two SSTRs, and that oligomerization of SSTRs is regulated by ligand binding by a selective process that is restricted to some (R5) but not other (R1) SSTR subtypes. We propose that induction by ligand of different oligomeric states of SSTRs represents a unique mechanism for generating signaling specificity not only within the SSTR family but more generally in the HHR family. PMID- 11880656 TI - Heterologous regulation of trafficking and signaling of G protein-coupled receptors: beta-arrestin-dependent interactions between neurokinin receptors. AB - Cells express multiple G protein-coupled receptors that are simultaneously or sequentially activated by agonists. The consequences of activating one receptor on signaling and trafficking of another receptor are unknown. We examined the effects of selective activation of the neurokinin 1 receptor (NK1R) on signaling and trafficking of the NK3R and vice versa. Selective agonists of NK1R and NK3R induced membrane translocation of beta-arrestins (beta-ARRs). Dominant negative beta-ARR(319-418) inhibited endocytosis of NK1R and NK3R. Whereas an NK1R agonist caused sequestration of NK1R with beta-ARR in the same endosomes, thereby depleting them from the cytosol, beta-ARRs did not prominently sequester with the activated NK3R and rapidly returned to the cytosol. In cells coexpressing both receptors, prior activation of the NK1R inhibited endocytosis and homologous desensitization of the NK3R, which was dose-dependently reversed by overexpression of beta-ARR1. Similar results were obtained in enteric neurons that naturally coexpress the NK1R and NK3R. In contrast, activation of the NK3R did not affect NK1R endocytosis or desensitization. Thus, the high-affinity and prolonged interaction of the NK1R with beta-ARRs depletes beta-ARRs from the cytosol and limits their role in desensitization and endocytosis of the NK3R. Because beta-ARRs are critical for desensitization, endocytosis, and mitogenic signaling of many receptors, this sequestration is likely to have important and widespread implications. PMID- 11880657 TI - A stilbene synthase from Japanese red pine (Pinus densiflora): implications for phytoalexin accumulation and down-regulation of flavonoid biosynthesis. AB - Stilbene synthase (STS) and chalcone synthase (CHS) are plant-specific polyketide synthases that play key roles in the stilbenoid and flavonoid biosyntheses, respectively. We have recently isolated from Pinus densiflora three STS cDNAs (PDSTS1, PDSTS2, and PDSTS3) and one CHS cDNA (PDCHSX). We then heterologously expressed these cDNAs in Escherichia coli and characterized their properties. An unusual STS isozyme, PDSTS3, lacks the common C-terminal extension of STS because of a frame-shift mutation and shows the highest pinosylvin-forming activity among the STSs tested. Pinosylvin was shown to be a potent inhibitor of PDCHSX (K(i) = 6 microM) as well as PDSTS2 (K(i) = 13 microM), which presumably maintains the balance between the stilbenoid and flavonoid biosyntheses. PDSTS3 was insensitive to product inhibition. We identified PDSTS3 in the pine seedlings as well as full length STS. The data provide evidence that PDSTS3 is involved in the potential regulation of the stilbenoid and flavonoid biosynthetic pathways in pine trees. PMID- 11880659 TI - Proceedings of the 2nd National Symposium on Medical and Public Health Response to BioTerrorism Public Health Emergency and National Security Threat. Washington, DC, USA. November 2000. PMID- 11880658 TI - Neural mechanisms of planning: a computational analysis using event-related fMRI. AB - To investigate the neural mechanisms of planning, we used a novel adaptation of the Tower of Hanoi (TOH) task and event-related functional MRI. Participants were trained in applying a specific strategy to an isomorph of the five-disk TOH task. After training, participants solved novel problems during event-related functional MRI. A computational cognitive model of the task was used to generate a reference time series representing the expected blood oxygen level-dependent response in brain areas involved in the manipulation and planning of goals. This time series was used as one term within a general linear modeling framework to identify brain areas in which the time course of activity varied as a function of goal-processing events. Two distinct time courses of activation were identified, one in which activation varied parametrically with goal-processing operations, and the other in which activation became pronounced only during goal-processing intensive trials. Regions showing the parametric relationship comprised a frontoparietal system and include right dorsolateral prefrontal cortex [Brodmann's area (BA 9)], bilateral parietal (BA 40/7), and bilateral premotor (BA 6) areas. Regions preferentially engaged only during goal-intensive processing include left inferior frontal gyrus (BA 44). The implications of these results for the current model, as well as for our understanding of the neural mechanisms of planning and functional specialization of the prefrontal cortex, are discussed. PMID- 11880660 TI - Confronting a world of infectious diseases. PMID- 11880661 TI - Biological weapons as a strategic threat. PMID- 11880663 TI - Reducing the bioweapons threat: international collaboration efforts. PMID- 11880662 TI - CDC's strategic plan for bioterrorism preparedness and response. AB - The Department of Health and Human Services (DHHS) has played a critical lead role over the past two years in fostering activities associated with the medical and public health response to bioterrorism. Based on a charge from Secretary Donna Shalala in 1998, the Centers for Disease Control and Prevention (CDC) is leading public health efforts to strengthen the nation's capacity to detect and respond to a bioterrorist event. As a result of our efforts, federal, state, and local communities are improving their public health capacities to respond to these types of emergencies. For many of us in public health, developing plans and capacities to respond to acts of bioterrorism is an extension of our long standing roles and responsibilities. These are stated in the CDC Mission Statement: to promote health and quality of life by preventing and controlling disease, injury, and disability, and the Bioterrorism Mission: to lead the public health effort in enhancing readiness to detect and respond to bioterrorism. CDC's infectious diseases control efforts are summarized below: --Initially formed to address malaria control in 1946; --Established the epidemic Intelligence Service in 1951; --Participated in global smallpox eradication and other immunization programs; --Estimated 800-1,000 + field investigations/year since late 1990s; - New diseases: Legionnaire's Disease, toxic shock syndrome, Lyme disease, HIV, hantavirus pulmonary syndrome, West Nile, etc. -- Today: focus on emerging infections and bioterrorism. Over the past 50 years, CDC has seen a decline in the incidence of some infectious diseases and an increase in some, whereas others continue to present on a more unpredictable basis (i.e., hantavirus). Outbreak identification, investigation, and control have been an integral part of what we do for more than 50 years. We estimate that 800 to 1,000 field investigations have occurred every year since the late 1990s. Today, however, we have a new focus on emerging infectious diseases and bioterrorism. PMID- 11880664 TI - Olympics 2000: preparing to respond to bioterrorism. PMID- 11880666 TI - Global awareness of disease outbreaks: the experience of ProMED-mail. PMID- 11880665 TI - International cooperation to prevent biological weapons research and development. PMID- 11880667 TI - "Hospital's full-up": the 1918 influenza pandemic. PMID- 11880668 TI - Treating the sick: capacity of the US Health Care System to respond to an epidemic. PMID- 11880669 TI - Hospital preparedness for bioterrorism. PMID- 11880670 TI - Mobilizing professional communities. PMID- 11880671 TI - Institutional networks: regional response to disasters. PMID- 11880672 TI - National disaster medical system. PMID- 11880674 TI - Challenges confronting public health agencies. PMID- 11880673 TI - International leadership in the control of biological weapons. PMID- 11880675 TI - Observations from the Top Off exercise. PMID- 11880676 TI - Understanding public response to disasters. PMID- 11880677 TI - How to vaccinate 30,000 people in three days: realities of outbreak management. PMID- 11880678 TI - Legal issues surrounding public health emergencies. PMID- 11880679 TI - Understanding media's response to epidemics. PMID- 11880680 TI - Epidemic response scenario: decision making in a time of plague. PMID- 11880681 TI - Envisioning worldwide disarmament. PMID- 11880682 TI - The problem of biological weapons: next steps for the nation. PMID- 11880683 TI - National leadership in confronting bioterrorism: 1. PMID- 11880684 TI - National leadership in confronting bioterrorism: 2. PMID- 11880686 TI - Software for signaling networks, electronic and cellular. AB - Cellular signaling networks have many similarities with electronic circuits. Therefore, the tools used to test and analyze electronic circuits can be adapted to analyze cellular signaling processes. Lok describes some of these signaling network analysis tools, their underlying principles, and their limitations. PMID- 11880687 TI - Ordered just so: lipid rafts and lymphocyte function. AB - Immunologists have long been occupied with the description of cellular activation signaling events that originate with the stimulation of multichain immunoreceptors at the cell surface. These signals are transmitted by a protein partner-signaling cascade through the cytoplasm to the nucleus, where they culminate in changes in gene expression, metabolic state, and entry into cell cycle. For T cells and B cells, these signaling cascades start with the ligation of the T cell receptor (TCR) and B cell receptor (BCR), respectively, and result in the recruitment and activation of related families of signaling molecules at the cell surface. Until recently, this gathering of signaling proteins was thought to occur within the featureless plasma membrane, a cellular organ that was envisioned as a boundary between the inner and outer components of the cell, but which contributed little to the signaling process. However, the past few years have seen the gradual realization that activation of signaling in lymphocytes takes place in and around specialized membrane subdomains called lipid rafts (also known as DIGs and GEMs). Here, we provide a brief overview of the analogous structures and compositions of lipid raft-associated signaling complexes in T cells and B cells, and the ways in which lymphocytes--and their pathogen adversaries--use lipid rafts to their benefit. PMID- 11880685 TI - How and why does the immunological synapse form? Physical chemistry meets cell biology. AB - During T lymphocyte (T cell) recognition of an antigen, a highly organized and specific pattern of membrane proteins forms in the junction between the T cell and the antigen-presenting cell (APC). This specialized cell-cell junction is called the immunological synapse. It is several micrometers large and forms over many minutes. A plethora of experiments are being performed to study the mechanisms that underlie synapse formation and the way in which information transfer occurs across the synapse. The wealth of experimental data that is beginning to emerge must be understood within a mechanistic framework if it is to prove useful in developing modalities to control the immune response. Quantitative models can complement experiments in the quest for such a mechanistic understanding by suggesting experimentally testable hypotheses. Here, a quantitative synapse assembly model is described. The model uses concepts developed in physical chemistry and cell biology and is able to predict the spatiotemporal evolution of cell shape and receptor protein patterns observed during synapse formation. Attention is directed to how the juxtaposition of model predictions and experimental data has led to intriguing hypotheses regarding the role of null and self peptides during synapse assembly, as well as correlations between T cell effector functions and the robustness of synapse assembly. We remark on some ways in which synergistic experiments and modeling studies can improve current models, and we take steps toward a better understanding of information transfer across the T cell-APC junction. PMID- 11880688 TI - Effects of physical conditioning on cardiac autonomic function in healthy middle aged women. AB - The purpose of this study was to determine whether moderate exercise training affects heart rate variability (HRV) and spontaneous baroreflex (SBR) function in healthy middle-aged women. Thirty-two sedentary women aged 40-59 yr were divided into four groups depending on reproductive state (premenopausal, PrM or postmenopausal, PoM, and training group (exercise or sedentary control group). The electrocardiographic R-R interval and systolic blood pressure (finger plethysmograph) were measured at rest during paced breathing at 16 breaths/min in the left lateral decubitus, sitting, and free standing postures, and during upright cycling at 40% maximal heart rate reserve (MHRR). After initial testing, the exercise groups underwent a 12 week walking program. After training, both exercise groups (PrM and PoM) had a longer R-R interval in all conditions, a higher SBR slope in the sitting and standing posture and lower sympathetic modulation in the standing posture, than the control groups and pre-training levels. During exercise, vagal modulation was higher and sympathetic modulation was lower in both exercise groups compared to pre-training values. Vagal modulation is increased following moderate intensity aerobic conditioning in middle-aged women. PMID- 11880690 TI - What is fatigue? AB - Fatigue and potentiation are two forms of force modulation. A general definition of fatigue is "a circumstance where less than the anticipated contractile response is obtained." Fatigue is associated with depressed Ca2+ release and possibly decreased Ca2+ sensitivity. Potentiation results from increased Ca2+ sensitivity due to regulatory light chain phosphorylation. Muscle fatigue and potentiation can coexist, making it difficult to quantify these processes. With repetitive 10Hz stimulation, the developed tension first increases, then decreases. Is fatigue present when developed tension first begins to decrease or when it falls below the developed tension of the first response? Intermittent incompletely fused tetanic contractions for which peak developed tension first decreases, then increases, is another unusual example of fatigue. A third example is when twitch contractions following a tetanic contraction decrease to a level below the pretetanic twitch amplitude, indicating that fatigue may have been coexistent with posttetanic potentiation. These observations illustrate the complexity of detecting fatigue, based on the simple, but commonly accepted definition presented above. Care must be taken in interpreting "before vs. after" contractile responses. Even when the contraction amplitude is greater than the initial response, there is no guarantee that mechanisms associated with fatigue are not present. PMID- 11880689 TI - Cellular and molecular basis of age-related sarcopenia. AB - Sarcopenia, the decline in muscle bulk and performance associated with normal aging, is an important component of frailty in the elderly. The gradual loss of both motor nerves and muscle fibers during senescence appears to be the major problem. Atrophy (especially in fast-twitch fibers) and impaired function of the surviving cells also contribute to sarcopenia. Although skeletal muscle has the capacity to regenerate itself, this process is not activated by the gradual age related loss of muscle fibers. The endocrine, autocrine, and paracrine environment in old muscle is less supportive of protein synthesis, reinnervation of muscle fibers, and satellite cell activation, proliferation, and differentiation. Lifelong exposure of DNA to free radical damage results in accumulation of somatic mutations in nerves and muscle fibers. Reduced protein synthesis leads to atrophy, and slower fractional protein turnover contributes to longer retention of proteins that may have been damaged by free radicals. Many genes are differentially expressed in young and old muscle, but additional research is needed to determine which of these genes have a significant role in the pathogenesis or adaptation to sarcopenia. PMID- 11880691 TI - The role of O2 supply in muscle fatigue. AB - It is well established that altering O2 delivery to contracting skeletal muscle affects human performance. In this respect, a reduced O2 supply (e.g., hypoxia) increases the rate of muscle fatigue, whereas increasing O2 supply (e.g., hyperoxia) reduces the rate of fatigue. Interestingly, the faster onset of fatigue in moderate hypoxia does not appear to be a consequence of mitochondrial O2 limitation because these effects occur at submaximal rates of O2 consumption for these conditions and at O2 tensions well above that which impairs mitochondrial O2 uptake in vitro. Alterations in O2 supply modulate the regulation of cellular respiration and may affect the onset of impaired Ca2+ handling with fatigue. Specifically, changes in O2 supply alter the coupling between phosphocreatine hydrolysis and O2 uptake in contracting muscles, which by determining the rate of inorganic phosphate (Pi) accumulation may affect Ca2+ release. Partial ischemia differs somewhat in that the reduction in force could be due to reduced O2 supply and/or impaired removal of metabolic by-products secondary to insufficient blood flow. Nonetheless, recent evidence shows a parallel decline and restoration of force with alterations in O2 supply but not blood flow alone during submaximal contractions. Furthermore, the causes of fatigue are similar when O2 is plentiful and when it is reduced. PMID- 11880692 TI - Muscle fatigue: the role of metabolism. AB - This paper examined the role of metabolites in causing muscle fatigue. Previous studies have shown that Pi (H2PO4-, HPO4-2) and H+ may be important factors in causing fatigue. A key question is the potential interaction between metabolic end-products and calcium related excitation-contraction coupling fatigue (ECC). An in vivo rat muscle model was used to measure tension development and metabolic end-products in response to electrical stimulation. Two stimulation protocols were used, high intensity stimulation followed by a medium intensity stimulation (High Group), and low intensity stimulation followed by a medium intensity stimulation (Low Group). Metabolic fatigue was based on concentrations of (H2PO4 measured with phosphorus magnetic resonance spectroscopy. ECC fatigue was measured as the fatigue in excess of metabolic fatigue, and as the relative decline of force at low compared to high stimulation frequencies. During the initial stimulation period, the High Group had greater metabolic fatigue (p < 0.001) and greater ECC fatigue (p = 0.007). During the second stimulation period and recovery, the High Group had no difference in metabolic fatigue (p = 0.07) and greater ECC fatigue (p = 0.015). These results present a method for determining the relative amounts of metabolic and ECC fatigue, and suggest that metabolites can increase the amount of ECC fatigue. PMID- 11880693 TI - Muscle fatigue: the role of intracellular calcium stores. AB - Force declines when muscles are used repeatedly and intensively and a variety of intracellular mechanisms appear to contribute to this muscle fatigue. Intracellular calcium release declines during fatigue and has been shown to contribute to the reduction in force. Three new approaches have helped to define the role of calcium stores to this decline in calcium release. Skinned fibre experiments show that when intracellular phosphate is increased the amount of Ca2+ released from the sarcoplasmic reticulum (SR) declines. Intact fibre experiments show that the size of the calcium store declines during fatigue and recovers on rest. Intact muscles which lack the enzyme creatine kinase, do not exhibit the usual rise of phosphate during fatigue and, under these conditions, the decline of Ca2+ release is absent or delayed. These results can be explained by the "calcium phosphate precipitation" hypothesis. This proposes that if phosphate in the myoplasm rises, it enters the SR and binds to Ca2+ as Ca2+ phosphate. The resultant reduction in free Ca2+ within the SR contributes to the reduced Ca2+ release during fatigue. PMID- 11880694 TI - Recurrent acute pancreatitis. Introduction. PMID- 11880695 TI - Aetiologies of recurrent acute pancreatitis: acute or chronic relapsing disease? PMID- 11880696 TI - Role of hereditary pancreatitis and CFTR gene mutations in the aetiology of acute relapsing pancreatitis of unknown origin. How are they important? PMID- 11880697 TI - Acute relapsing pancreatitis. Congenital variants: diagnosis, treatment, outcome. PMID- 11880698 TI - Sphincter of Oddi dysfunction: diagnosis and treatment. PMID- 11880699 TI - What should be done with idiopathic recurrent pancreatitis that remains 'idiopathic' after standard investigation? PMID- 11880700 TI - Acute recurrent pancreatitis and dysfunction of the sphincter of Oddi: comparison between invasive and non-invasive techniques. PMID- 11880701 TI - Is secretin magnetic resonance cholangio-pancreatography an effective guide for a diagnostic and/or therapeutic flow-chart in acute recurrent pancreatitis? PMID- 11880702 TI - High-dose cyclophosphamide for aplastic anemia and autoimmunity. AB - High-dose cyclophosphamide was developed as a conditioning regimen for allogeneic bone marrow transplantation. Later, it was discovered that high-dose cyclophosphamide spares early hematopoietic stem cells because of their relatively high levels of the enzyme aldehyde dehydrogenase; thus, high-dose cyclophosphamide is a potent immunosuppressive agent, but nonmyeloablative. Recent reports demonstrate that high-dose cyclophosphamide without bone marrow transplantation induces durable treatment-free remissions in severe aplastic anemia and a variety of other autoimmune disorders; however, there is lingering concern about the safety of this approach. PMID- 11880703 TI - Bone marrow transplantation for multiple myeloma: where we are today. AB - Multiple myeloma is incurable with standard chemotherapy. Autologous transplantation appears to offer a modest survival advantage over standard dose chemotherapy, but most patients subsequently relapse. Through the induction of graft-versus-tumor activity, allogeneic bone marrow transplantation can lead to long-term disease-free survival, and cure in some patients with myeloma. Transplant-related mortality after allogeneic bone marrow transplantation is high. Many patients are ineligible for this approach because of advanced age, comorbid illnesses, and extensive previous chemotherapy. Ongoing investigations endeavor to reduce regimen-related mortality through nonmyeloablative preparative regimens while maintaining immunologic antitumor activity through donor lymphocytes, which have significant graft-versus-myeloma activity. Early reports demonstrate lower rates of transplant related mortality; however, graft-versus host disease rates are high and can preclude the administration of graded donor lymphocyte infusions, which may optimize the therapeutic index of graft-versus host reactivity. PMID- 11880704 TI - Autologous stem cell transplantation for adult acute leukemia. AB - Autologous stem cell transplantation using marrow or peripheral blood is routinely used to consolidate patients with acute myelocytic leukemia in complete remission. The situation is less clear for adult acute lymphocytic leukemia in which results achieved with all strategies are disappointing. In acute myelocytic leukemia, autografts should be done in patients with good and standard risk factors. Patients with high-risk acute myelocytic leukemia defined by poor cytogenetics or failure to achieve remission with the first induction course, should proceed to allogeneic stem cell transplantation with the best available human leukocyte antigen-identical donor (family or unrelated), and the nature of the conditioning regimen (myelo-ablative or non-myelo-ablative) should be decided in relation to age, and the patient's clinical condition. Results of autografting in acute myelocytic leukemia rely strongly on the quality of the graft. Higher doses of infused stem cells translate into lower relapse rates. Marrow purging with cyclophosphamide derivatives also diminishes the relapse incidence. Autologous stem cell transplantations using peripheral blood are presently preferred to marrow as the source of stem cells, but an aggressive prior in vivo purge (high-dose consolidation course(s) before cytaphereses) is then mandatory. In good-risk acute myelocytic leukemia, autografting is superior to high-dose ARA C; in standard-risk acute myelocytic leukemia, both are supposedly equivalent. There is no prospective randomized study testing the two approaches in the good standard-risk population. We presently test the combination of marrow and blood both purged by mafosfamide. In adult acute lymphocytic leukemia, good-risk patients get the best benefit from autografting over conventional chemotherapy. Maintenance chemotherapy after transplant is likely to bring benefit. Research in progress aims at facilitating access of the largest number of patients to autografting and at introducing posttransplant immunomodulation maneuvers such as tumor vaccination. PMID- 11880705 TI - Umbilical cord blood transplantation: current state of the art. AB - Human leukocyte antigen-matched sibling donor umbilical cord blood transplantation, or 0-2 human leukocyte antigen mismatched unrelated donor umbilical cord blood, is now considered an acceptable alternative to the use of bone marrow as a source for hematopoietic stem cells for pediatric hematopoietic stem cell transplantation, and is being investigated in adults. Major advantages of umbilical cord blood include the speed of availability compared with unrelated donor bone marrow, and tolerance of 1-2 human leukocyte antigen mismatch, which offers the opportunity to extend the donor pool. Umbilical cord blood transplantation is associated with durable engraftment and low incidence of severe graft-versus-host disease, even in the 1-2 human leukocyte antigen mismatched setting. Clinical experience has established the importance of graft cell dose in determining engraftment and survival in unrelated donor umbilical cord blood transplantation. More recently, the influence of human leukocyte antigen on outcome has become apparent. This review outlines the state of the art of umbilical cord blood transplantation, with emphasis on practical considerations in umbilical cord blood selection, and describes current research directions for this hematopoietic stem cell source. PMID- 11880706 TI - Hematopoietic stem cell transplantation in Hodgkin disease. AB - Intensive therapy and autologous stem cell transplantation represent the standard of care in most patients whose Hodgkin disease has not been cured with conventional chemotherapy. With more prolonged follow-up of autografted patients, the problems with autologous stem cell transplantation are clear. In particular, recurrent disease and late transplant-related complications limit the effectiveness of this approach. A number of autologous stem cell transplantation studies have reported prognostic factors that will help identify patients at high risk for relapse. Several new approaches for decreasing recurrence rates are discussed, including novel immune strategies and re-evaluation of allogeneic stem cell transplantation. Although the risk of secondary malignancy and other causes of late morbidity and mortality after autologous stem cell transplantation is relatively low, current studies contribute to understanding of the pathogenesis of these complications and may diminish their impact in the future. PMID- 11880707 TI - Translational oncogenomics: toward rational therapeutic decision-making. AB - The wealth of data in basic science and translational research may often represent a conundrum for clinical oncologists, who need to selectively consider this abundant information and translate it into therapeutic decisions. For the sake of simplicity, we have classified the multiplicity of genetic abnormalities in five repertoires that are rapidly assessable and useful for stratification in clinical trials: allelic imbalance, aberrant promoter methylation, gene mRNA overexpression, microtubule alterations, and polymorphisms. Allelic imbalance refers to chromosomal instability, which is a major feature of cancer, and innovative techniques used in colorectal cancer should also be implemented in lung cancer. Epigenetic changes (variations in transcription levels) have been extensively studied in non-small cell lung cancer. Methylation techniques have shown that these epigenetic changes commonly occur at the same frequency in numerous genes, both well-known ( FHIT, APC, p16 ) and recently discovered ( TMS1, RASSF1 ) in non-small cell lung cancer and in breast cancer. Innovative techniques like quantitative polymerase chain reaction can determine gene expression profiles, mainly overexpression of mRNAs, which may be related to resistance to specific cytotoxic drugs. In the near future, we hope these profiles can be used to individualize chemotherapy. Multiple microtubule alterations related to overexpression of different genes can also be used to predict response to taxanes and Vinca alkaloids. Finally, the assessment of polymorphisms could enable us to understand their functional consequences in chemotherapy response. PMID- 11880708 TI - Inductive and adjuvant treatment strategies for localized nonsmall cell lung cancer in operable and inoperable patients. AB - For many years, surgery has been considered the best treatment modality for resectable nonsmall cell lung cancer. However, survival has not increased over the last few decades. The reported 5-year survival rates for the early stages (stage I and II) show failures of 35 to 65%. Therefore, a combined modality approach has been taken in patients with N2 (lymph node) disease to improve results. These adjuvant treatments, consisting of chemotherapy, radiotherapy, or both, have yielded mixed responses. Adjuvant radiation therapy alone has shown a decrease in local recurrence but no significant improvement in survival. Randomized trials of adjuvant chemotherapy have shown mixed results, and large, randomized trials are currently being evaluated. Combinations of radiation and chemotherapy in the adjuvant setting have failed to show a survival benefit so far. In the neoadjuvant setting, chemotherapy has aroused great attention because it has significantly improved survival in patients with locally advanced nonsmall cell lung cancer (N2 disease). This result led to the use of neoadjuvant chemotherapy in early-stage lung cancer. Multinational and multi-institutional studies have been initiated recently. In the future, researchers will have clear insight into the effects of the classical chemotherapy regimen as induction therapy in the early stages of nonsmall cell lung cancer. Tyrosine kinase inhibitors, antisense therapies, and antiangiogenesis therapies are the most promising developments of the last few years. Implementation of these agents is expected to have great impact on the survival in patients with nonsmall cell lung cancer. PMID- 11880709 TI - Chemoprevention strategies for non-small cell lung cancer. AB - Among all cancers, lung cancer has the highest rate of mortality in the western world, both in men and women. The poor lung cancer survival rates argue powerfully for new approaches to control this disease, such as chemoprevention, which has been defined as the use of agents that reverse, suppress, or prevent lung carcinogenesis. Over 80% of lung cancers are attributed to tobacco and carcinogens from cigarette smoke, which unquestionably links nicotine addiction and lung cancer. Epidemiologic studies show that not more than 15% of heavy smokers will ultimately develop lung cancer. That 85% of heavy smokers will not develop lung cancer points to differences in susceptibility. Diet and genetically determined factors seem to play an important role in modulating individual susceptibility and are closely linked to the chemoprevention approach. Despite encouraging data from experimental and epidemiologic studies, the evidence for a positive effect in the human situation is still controversial. However, recent developments in molecular biology and increased insight in lung carcinogenesis have led to the potential of more specifically targeted intervention and optimism for new, successful chemopreventive approaches. PMID- 11880710 TI - Chemotherapy in advanced nonsmall cell lung cancer: indication, intensity, and duration. AB - Platinum-based combination and single-agent chemotherapy have become accepted as treatments for locally advanced and metastatic nonsmall cell lung cancer as a consequence of improved survival, quality of life, and symptom control compared with best supportive care. However, it is clear that a therapeutic plateau has been reached with current combinations requiring a re-evaluation of strategies to improve clinical outcomes. Dose intensification may offer one way in which to achieve better results, as may extension of the duration of treatment. The evidence suggests that dose intensification is a useful tool, and that its use in combination with markers of treatment duration and cumulative dose may help to maximize results from current active drug combinations. PMID- 11880711 TI - Individualized chemotherapy for elderly patients with nonsmall cell lung cancer. AB - Approximately one third of all patients with nonsmall cell lung cancer (NSCLC) are over the age of seventy. Elderly patients tolerate chemotherapy poorly because of impaired organ function and comorbidities. For this reason, these patients are often not considered eligible for aggressive cisplatin-based chemotherapy. A multidimensional geriatric evaluation is important to plan appropriate treatments. At present, there are no indications for adjuvant and neoadjuvant chemotherapy. Combined chemoradiotherapy in locally advanced disease increases toxicity and seems determine no survival advantage as compared with radiation therapy alone. In advanced disease, single-agent vinorelbine proves to be active and well-tolerated, and compared with best supportive care, improves survival and perhaps quality of life. Gemcitabine is active and also well tolerated. Taxanes are in advanced phase of evaluation. A phase III randomized trial showed that polychemotherapy with gemcitabine and vinorelbine does not improve any outcome as compared with single-agent chemotherapy with vinorelbine or gemcitabine. In clinical practice, single-agent chemotherapy should remain the standard treatment. The choice of the drug should be based on the toxicity profile of each drug and type of comorbid conditions. In the near future, new therapeutic strategies and biologic agents could improve present results. PMID- 11880712 TI - European approach to the treatment of malignant melanoma. AB - The European approach to the treatment of each stage of malignant melanoma can be characterized as cautious, avoiding unwarranted mutilation or toxicity, because phase III trials have demonstrated that an aggressive approach in surgical management, adjuvant therapy, and treatment of stage IV disease has met with little success. Phase III trials have demonstrated that wide margins, elective lymph node dissections, and prophylactic isolated limb perfusions bring no survival benefit. Primary melanoma is excised with a margin of 1 cm to maximally 2 cm and primary closure as a rule. There is no standard adjuvant therapy. High dose interferon treatment is practiced only sporadically in Europe because its high toxicity profile and an unclear long-term impact on survival are not popular. Long-term nontoxic lower-dose interferon regimens and vaccines are currently being explored. Phase III trials have shown that highly toxic polychemotherapy or biochemotherapy has not produced a survival benefit over simple treatment with dacarbazide alone. In Europe biochemotherapy is being abandoned and various less toxic or nontoxic approaches with vaccines and antiangiogenic agents are under study. PMID- 11880713 TI - Cutaneous toxicities of cancer therapy. AB - This review highlights the cutaneous side effects associated with the administration of chemotherapy and discusses the management of these conditions. Rapidly growing cells are the targets of chemotherapy, so the skin, hair follicles, and nail matrix are frequently affected by chemotherapy. Chemotherapy skin reactions are more likely toxic than allergic reactions. The most common cutaneous reactions are alopecia, hyperpigmentation, hand--foot syndrome, radiation recall, hypersensitivity, extravasation injuries, and nail dystrophies. While these side effects are generally not life threatening, they can be a source of significant distress to patients, especially alopecia. PMID- 11880714 TI - Update on the use of sentinel node biopsy in patients with melanoma: who and how. AB - This review highlights the utility of sentinel lymph node biopsy for melanoma. The histopathologic status of the sentinel lymph node is acknowledged as the most powerful indicator of prognosis. Changes by the American Joint Committee for Cancer have not only incorporated the importance of ulceration in cutaneous melanoma, but have focused on the number of metastatic lymph nodes as well as whether the disease is macro- or micrometastatic. Specific technical aspects of the sentinel lymph node procedure are discussed, with emphasis on practical guidelines to minimize incorrect nodal staging. PMID- 11880715 TI - The role of surgery for patients with metastatic melanoma. AB - When deciding to perform a resection for metastatic melanoma one should first decide whether the intent of the procedure is curative or palliative. When the resection is palliative, the success of surgical treatment will depend on the presence of identifiable symptoms, the morbidity of the procedure, the course of the disease, and the ability to communicate the treatment goals among surgeon, patient, and family. When the resection is curative, survival will depend on the ability of the surgeon to select patients with a pattern of recurrence suggestive of less aggressive tumor biology. Factors generally found predictive of improved survival, and therefore reflective of tumor biology, include longer disease-free interval, fewer numbers of metastases, and the ability to obtain a complete resection. Resection of metastases in patients who recur within one-year, who present with multiple lesions, and who present with disease that cannot be completely resected, will not result in long-term survival. PMID- 11880716 TI - Genetic basis of glaucoma. AB - The application of molecular genetic techniques to the study of glaucoma has accelerated greatly during the past few years. In addition to localizing and identifying genes for specific types of glaucoma, researchers have begun to characterize the gene products and investigate molecular mechanisms involved in glaucoma. Much research has been focused on the gene expression, protein processing, and mutations of MYOC/TIGR, which is associated with both juvenile and adult-onset primary open angle glaucoma. Investigations of other glaucoma related genes, such as PITX2, FOXC1, and CYP1B1, are enabling a better understanding of anterior segment development and its relation to glaucoma. PMID- 11880717 TI - Current understanding of neuroprotection in glaucoma. AB - The most common optic neuropathy is glaucomatous optic neuropathy, distinguished by a distinctive and progressive excavation of the optic nerve head without significant pallor of the remaining neuroretinal rim. Neuroprotection is a novel strategy for treating disorders that affect the nervous system by preventing death of neurons. In glaucomatous optic neuropathy, the neurons that die are retinal ganglion cells. This article reviews the recent basic science relevant to neuroprotection, particularly with respect to retinal ganglion cell death in glaucomatous and other optic neuropathies. PMID- 11880718 TI - Optic nerve and retinal nerve fiber layer analyzers in glaucoma. AB - There is mounting evidence that retinal nerve fiber layer (RNFL) loss precedes detectable visual field loss in early glaucomatous optic neuropathy. However, examination and photography of the RNFL is a difficult technique in many patients, particularly older individuals, and eyes with small pupils and media opacities. It is subjective, qualitative, variably reproducible, and often unreliable. Furthermore, optic nerve head and RNFL photography is time consuming, operator dependent, has limited sensitivity and specificity, and requires storage space. Imaging technologies have emerged which enable clinicians to perform accurate, objective, and quantitative measurements of the RNFL and optic nerve head topography. There is good agreement between such measurements and clinical estimates of optic nerve head structure and visual function. The reproducibility of these instruments suggests that they have the potential to detect structural change over time. This report will review the technological principles, reproducibility, sensitivity and specificity, capacity to detect glaucomatous progression, and limitations of currently available ocular imaging technologies. PMID- 11880719 TI - Recent developments in automated perimetry in glaucoma diagnosis and management. AB - Recently, there have been several new developments in automated perimetry that have contributed to enhanced diagnosis and management of glaucoma. This paper will briefly review four of the latest advances in automated perimetry: (1) efficient test strategies that reduce the testing time and variability of automated perimetric testing, in particular, the Swedish Interactive Threshold Algorithm (SITA) and Tendency Oriented Perimetry (TOP) test strategies will be described; (2) Frequency Doubling Technology (FDT) perimetry, which has been shown to be a rapid, effective method of detecting glaucomatous visual field loss; (3) Short Wavelength Automated Perimetry (SWAP), which has demonstrated the ability to predict the onset and progression of glaucomatous visual field deficits; (4) The Multifocal Electroretinogram (mfERG) and the Multifocal Visual Evoked Potential (mfVEP), which provide an objective measurement of the visual field. Each of these techniques has presented distinct advantages for the diagnosis and management of glaucoma. PMID- 11880720 TI - How clinical trial results are changing our thinking about target pressures. PMID- 11880721 TI - Review of recent advancements in the understanding of primary angle-closure glaucoma. AB - Primary angle-closure glaucoma is a leading cause of blindness worldwide. However, the terminology used for angle-closure in the literature is inconsistent, with inappropriate emphasis on symptomatology. A new nomenclature for primary angle-closure glaucoma focuses on the presence of end-organ damage and limits the use of the term "glaucoma" only for people who have suffered injury to the optic nerve. This review describes the various modalities of treatment for primary angle-closure glaucoma. The role and limitations of laser peripheral iridotomy in the management of the different forms of the disease are summarized. Recent developments have led to improvements in the understanding of the epidemiology, clinical course, and treatment of the condition. PMID- 11880722 TI - Selective laser trabeculoplasty: a new treatment option for open angle glaucoma. AB - Selective laser trabeculoplasty (SLT) is a safe and effective treatment modality for lowering the intraocular pressure in patients with open angle glaucoma. The preservation of the trabecular meshwork architecture and the demonstrated efficacy in lowering intraocular pressure makes the SLT a reasonable and safe alternative to argon laser trabeculoplasty. In addition, SLT is a potentially repeatable procedure because of the lack of coagulation damage to the trabecular meshwork and the demonstrated efficacy in patients with previously failed argon laser trabeculoplasty treatment. Furthermore, SLT can be considered as a primary treatment option in patients who cannot tolerate or are noncompliant with their glaucoma medications, while not interfering with the success of future surgery. Due to its nondestructive properties and low complication rate, SLT has the potential to evolve as an ideal first-line treatment in open angle glaucoma. PMID- 11880723 TI - Management of refractory glaucoma in childhood. AB - Glaucoma in children is characterized by marked intraocular pressure (IOP) elevation with resultant atrophy of the optic nerve and loss of retinal ganglion cells. In very young children, secondary expansion of the globe with damage to anterior segment structures, such as the cornea and zonule, often occurs. Permanent, severe visual dysfunction may result from optic nerve damage as well as from amblyopia arising from anisometropia and corneal opacification. The treatment of childhood glaucoma often involves surgery. Goniotomy and trabeculotomy remain the first line surgical procedures for open-angle glaucoma in children. Trabeculectomy with adjunctive antifibrosis therapy, aqueous shunt surgery, and cyclodestructive procedures are undertaken when angle surgery fails to control the IOP or is unlikely to succeed. The choice of surgical procedure is individualized according to factors such as the age of the patient, the specific type of glaucoma, the number of prior surgical procedures, and the visual potential of the eye. Achieving and maintaining an adequate IOP to prevent progressive optic nerve damage, avoiding complications, and preserving vision are the goals that must be considered in deciding on a surgical plan. PMID- 11880724 TI - Management of glaucoma after retinal detachment surgery. AB - Secondary glaucoma may complicate retinal detachment surgery. Intraocular pressure (IOP) elevation has been described after scleral buckling procedures and vitrectomy with intravitreal injection of gas or silicone oil. Angle-closure glaucoma after scleral buckling develops because of congestion and anterior rotation of the ciliary body. Medical therapy and laser iridoplasty are usually successful in controlling IOP, but the presence of conjunctival scarring and recession and retinal hardware after scleral buckling procedures can make surgical management challenging. Intravitreal injection of expansile gases like sulfur hexafluoride (SF6) and perfluoropropene (C3F8) may produce secondary angle closure glaucoma with or without pupillary block. Aspiration of a portion of the intraocular gas may be needed, especially if IOP is elevated to a level that may compromise ocular perfusion. Glaucoma also can develop after intravitreal injection of silicone oil secondary to pupillary block, inflammation, synechial angle closure, rubeosis iridis, or migration of emulsified or nonemulsified silicone oil into the anterior chamber. A prophylactic inferior iridectomy at the time of surgery serves to prevent pupillary block. Patients with medically uncontrolled glaucoma after silicone oil injection may require oil removal with or without concurrent glaucoma surgery. PMID- 11880725 TI - Nonpenetrating filtering surgery. AB - Trabeculectomy is currently the standard filtration procedure for surgical treatment of glaucoma. This technique has advantages over full thickness procedures but can cause early postoperative complications such as hyphema, shallow or flat anterior chamber, hypotony with the risk of choroidal detachment, and maculopathy. Nonpenetrating procedures are alternatives to trabeculectomy with the advantage of minimizing the risk of postoperative complications related to hypotony. The two major variations of nonpenetrating glaucoma surgery are nonpenetrating deep sclerectomy and viscocanalostomy. Techniques and results are discussed in this paper. PMID- 11880726 TI - Adolescents with cancer: access to clinical trials and age-appropriate care. AB - Many adolescents with cancer are denied the benefits of protocol inclusion and optimal supportive care. More than 70% of older adolescent patients are not treated at institutions representing pediatric cooperative groups and are not enrolled in clinical trials, despite a demonstrated survival advantage for patients registered in protocols. Failure to refer adolescent cancer patients to specialized pediatric oncology treatment centers also has an impact on their quality of care. The challenge before both the medical and pediatric health care communities is to assure adolescent patients the benefits of inclusion in clinical trials and treatment in age-appropriate settings. PMID- 11880727 TI - Recent advances in Wilms tumor genetics. AB - The past decade has witnessed substantial growth in our knowledge of the genes and loci that are altered in Wilms tumor. Although Wilms tumor was one of the original paradigms of Knudson's two-hit model of cancer formation, it has become apparent that several genetic events contribute to Wilms tumorigenesis. Recent research has identified targets and regulators of the first Wilms tumor gene, WT1, has uncovered several candidate genes at the second Wilms tumor locus, WT2, and has identified two familial Wilms tumor loci, FWT1 and FWT2. The recent discovery of activating beta-catenin mutations in some Wilms tumors has also implicated the Wnt signaling pathway in this neoplasm. Recurrent abnormalities of other loci, including 16q, 1p, and 7p, have indicated that these sites may harbor Wilms tumor genes. An enhanced understanding of these and other genetic lesions will provide the foundation for novel targeted Wilms tumor therapies. PMID- 11880728 TI - Gene therapy for the hemophilias. AB - Recent advances in the field of gene transfer are producing tantalizing results suggesting that the potential to correct disease at a molecular level may be at hand. Genetic correction of the hemophilias--bleeding disorders that stem from the deficiency of functional factor VIII or IX--represent models for the development of a basic understanding of how gene therapy will be achieved. The goals for hemophilia gene transfer are to produce therapeutic amounts of the coagulant protein while minimizing an immune response or antibody inhibitor. This requires the use of nontoxic vectors to deliver genes that express the protein in a functional form for the life of the patient. Based on a scientific understanding of the molecular and cellular defects leading to the bleeding phenotype, gene transfer studies at the laboratory and clinic have produced exciting results. The author here provides a critical assessment of the state of hemophilia gene transfer and its relevance to the field as a whole. PMID- 11880729 TI - Novel conditioning regimens and nonmyeloablative stem cell transplants. AB - Allogeneic stem cell transplantation has proven an effective means of treatment for a variety of malignant and nonmalignant conditions. Stem cell transplantation has traditionally been preceded by the patient receiving high (myeloablative) doses of chemotherapy and total body irradiation, which have been associated with significant transplant-related morbidity and mortality. This finding has precluded some older or heavily pretreated patients from being offered stem cell transplantation and has made other populations, such as those with nonmalignant conditions, reluctant to consider this therapy. Preclinical studies demonstrated that engraftment of donor stem cells could be achieved with nonmyeloablative, but immunosuppressive, preparative regimens that are significantly less toxic. Preliminary clinical studies have shown that patients previously ineligible for allogeneic stem cell transplantation can frequently be safely transplanted and that an immune-mediated graft-versus-malignancy effect plays an important role in the antitumor efficacy of this treatment. The potential to use this nonmyeloablative approach in nonmalignant disorders in which regimen-related toxicity must be minimized is of particular interest in children. PMID- 11880730 TI - Advances in the genetics and treatment of von Willebrand disease. AB - von Willebrand disease (VWD) is a bleeding disorder caused by quantitative (type 1 and 3) or qualitative (type 2) defects of von Willebrand factor (VWF). The mechanisms of most inherited VWD types have been recently elucidated by genetic and molecular diagnosis, but the phenotypic tests based on measurements of plasma and platelet VWF, the ability of VWF to interact with its platelet receptor, and the analysis of the multimeric composition of VWF are always essential to identify patients with different VWD subtypes. The aim of treatment is to correct the dual defects of hemostasis, ie, abnormal coagulation expressed by low levels of factor VIII (FVIII) and abnormal platelet adhesion expressed by prolonged bleeding time (BT). Desmopressin is the treatment of choice in most patients with type 1 and type 2 VWD, who account for 60 to 70% of cases. In type 3 and in some severe forms of type 1 and type 2 VWD, desmopressin is not effective, and it is necessary to resort to plasma concentrates containing FVIII and VWF. Treated with virucidal methods, these concentrates are effective and currently safe, but they do not always correct the BT defect. Platelet concentrates or desmopressin can be used as adjunctive treatments when poor correction of the BT after concentrates is associated with continued bleeding. PMID- 11880731 TI - Marfan syndrome: orthopedic and genetic review. AB - Marfan syndrome is an autosomal dominant disorder of connective tissue that affects the cardiac, eye, and skeletal systems. More than 135 mutations have been identified in the fibrillin-1 gene, localized on chromosome 15q21.1 [corrected] and responsible for the clinical manifestations of Marfan syndrome. The major orthopedic manifestations of Marfan syndrome include scoliosis, chest wall deformity, dural ectasia, joint hypermobility, and acetabular protrusion. In addition, decreased bone mineral density has been reported in patients with Marfan syndrome. This review summarizes recent developments in the genetic and orthopedic aspects of Marfan syndrome. Increased practitioner awareness of the clinical features associated with Marfan syndrome may facilitate earlier diagnosis and optimize patient treatment. PMID- 11880732 TI - Common pediatric hand fractures. PMID- 11880733 TI - Current treatment of hemophilic arthropathy. AB - Hemophilic arthropathy occurs in all patients with severe and moderate hemophilia A and B in early adolescence after repeated bleeding in a major joint unless treated with replacement of the missing factor. Regular infusions of recombinant factor or treated plasma derived factor given prophylactically to prevent spontaneous bleeding are recommended for all children to maintain a plasma factor level of >1%. Recombinant factor product or treated plasma derived product should be used. Prophylaxis should begin when bleeding occurs repeatedly and is superior to on-demand therapy. Hypertrophied synovium should be removed surgically or with a sclerosing agent, either radioactive or chemical material, to impede further cartilaginous and bony deterioration. Arthroplasty of the knee and hip have been successful in reducing pain and loss of motion when other efforts to control synovial hypertrophy fail. PMID- 11880734 TI - Orthopedic management of the muscular dystrophies. AB - Muscular dystrophy is a collective group of inherited, noninflammatory, progressive muscle wasting diseases. The initial pathologic feature is an abnormality in the genetic code for dystrophin or one of its associated glycoproteins, which leads to the various clinical syndromes. Despite minor variations between the different types, all muscular dystrophies have in common progressive muscle weakness, which is best typified by Duchenne muscular dystrophy. The weakness occurs in a proximal to distal direction and can compromise ambulatory status as well as cardiopulmonary function. Additionally, structural soft tissue contractures and spinal deformities may develop from poor posturing secondary to the progressive muscle weakness and imbalance. The rapidly developing scoliosis and its associated pelvic obliquity can even compromise sitting. Recent advances in molecular biology and gene therapy research raise the hope for a cure for muscular dystrophy in the near future. Until that time, however, the role of orthopedic surgeons in treating patients with muscular dystrophy is to preserve or prolong their functional status for as long as possible. This can be achieved by physical therapy, bracing, soft tissue releases for joint contractures, and early stabilization of the spine. PMID- 11880735 TI - Discoid lateral meniscus in children. AB - The discoid meniscus is the most common abnormal meniscal variant in children. It is most likely a congenital deviation and usually occurs laterally. The discoid shape results in greater coverage of the tibia and usually is associated with increased thickness of the meniscus that may lead to abnormal shearing forces across the knee joint. The Watanabe classification divides this anomaly into three distinct types: complete, incomplete, and Wrisberg ligament. The complete and incomplete types are often asymptomatic; unless symptomatic, they generally should be left alone. Tears should be treated with resection back to a stable rim. The Wrisberg ligament type is a hypermobile meniscus secondary to a lack of posterior tibial attachment. This type may or may not be associated with an abnormal shape and may appear in childhood as a classic snapping knee syndrome. These children complain of intermittent popping and snapping within the knee that clinically manifests as a dramatic audible and visible adjustment of the knee with each flexion and extension. Treatment of a symptomatic Wrisberg ligament type requires surgical repair of the posterior disruption. Saucerization of the remaining meniscus may be required to protect the repair from abnormal shear forces. PMID- 11880736 TI - Internet resources for pediatric infectious diseases. PMID- 11880737 TI - Dengue virus infections. AB - Dengue is the most important arthropod-borne viral disease of public health significance. Its geographic distribution includes more than 100 countries worldwide, where more than 2.5 billion people are at risk for dengue infections. Most people will have asymptomatic infections, but the disease manifestations range from an influenza-like disease known as dengue fever to a severe, sometimes fatal disease characterized by hemorrhage and shock, known as dengue hemorrhagic fever/dengue shock syndrome. Dengue fever and dengue hemorrhagic fever/dengue shock syndrome are caused by the dengue viruses (dengue-1, dengue-2, dengue-3, and dengue-4) transmitted from viremic to susceptible humans mainly by the bites of Aedes aegypti. There is no specific management of dengue infections, no vaccine is commercially available, and vector control is the only alternative for stopping the spread of the disease. Knowledge of several aspects of dengue infections, especially of diagnosis and vaccine development, is continuously evolving, but several issues are still unresolved. PMID- 11880738 TI - West Nile virus in the United States. AB - In the late summer of 1999, the first known cases of West Nile virus infection in the Western Hemisphere were recorded in New York City. These first cases were the hallmarks of an outbreak of West Nile virus infection that resulted in 7 deaths among 62 confirmed cases and an estimated 8200 asymptomatic to mild infections among residents and visitors in Queens, New York. This article reviews West Nile virus and its spread in the United States since its introduction in 1999. PMID- 11880739 TI - Transmission of HIV-1 from mother to infant. AB - In the developed world, antiretroviral therapy (ART) administered to the mother during pregnancy and intrapartum and to the infant in the neonatal period has resulted in a reduction of the overall risk of vertical transmission of HIV-1 to approximately 8%. In some settings, ART combined with cesarean section and a reduction in duration of ruptured membranes has resulted in a further lessening of risk to levels < or = 2 percent. A number of less expensive and greatly abbreviated ART regimens, useful for application in resource poor settings, also have resulted in reductions of mother-to-infant transmission of HIV-1 by 33 to 50% compared to baseline. A multitude of studies have shown these drugs to be safe for mothers, fetuses, and newborns. Breastfeeding seems to represent a risk factor that adds to the risk of vertical transmission, especially in infants who are fed a combination of breastmilk and other liquids and solids. Studies designed to assess the possible benefits of treating genital ulcer disease, chorioamnionitis, mastitis, and malnutrition in HIV-infected women, and of applying antiseptic washes to the cervix and vagina during labor, are in progress. PMID- 11880740 TI - Management of acute bacterial rhinosinusitis. AB - Acute bacterial rhinosinusitis is an infection of the nasal epithelium and paranasal sinus mucosa, usually caused in children by Streptococcus pneumoniae, Haemophilus influenzae, Moraxella catarrhalis, and, less frequently, group A Streptococcus species. The clinical diagnosis is based on daytime cough that may be worse at night or purulent rhinorrhea, or both, lasting at least 10 days, often worsening after a period of initial improvement after initial symptoms of the common cold, and often associated with facial or dental pain, facial fullness, or swelling, headache, and fever. Sinusitis is diagnosed clinically; radiographic evaluation is not indicated for diagnosis. When the disease persists despite treatment, or is complicated by potential intracranial or orbital extension, CT is the preferred imaging modality. Initial therapy should be amoxicillin in a high dosage (80-90 mg/kg/day). Treatment is generally for 10 to 14 days and for at least 7 days beyond the time of substantial improvement in symptoms. Complications of acute bacterial rhinosinusitis in children are rare. PMID- 11880742 TI - Cold-adapted influenza vaccines. AB - Because of the health impact of complicated disease from influenza in children, the role played by children in spreading influenza, and the limitations of inactivated influenza vaccine in this population, there has been ongoing effort to identify a more effective and acceptable vaccine. This review will focus on the development, limitations, and potential of a leading candidate vaccine studied in children. This is a live, attenuated, intranasally administered cold adapted influenza vaccine. PMID- 11880741 TI - Use of myeloid colony-stimulating factors in neonates with septicemia. AB - Bacterial sepsis is a major cause of neonatal morbidity and mortality. Successful management of neonatal sepsis requires early diagnosis, appropriate antimicrobial treatment, and aggressive intensive care. However, even when steps are taken appropriately, mortality rates can be high, particularly among certain subgroups, such as extremely preterm neonates and neonates with neutropenia. Multiple factors contribute to the increased susceptibility of neonates to infection, including developmental quantitative and qualitative neutrophil defects. Studies of infected animal and human neonates suggest that the use of recombinant human granulocyte colony stimulating factor (rhG-CSF) or recombinant human granulocyte macrophage colony stimulating factor (rhGM-CSF) can partially counterbalance these defects and thereby reduce morbidity and mortality. However, the body of clinical evidence is currently not sufficient to recommend rhG-CSF or rhGM-CSF administration confidently as routine adjunctive treatment for neonates with sepsis. PMID- 11880743 TI - Case report: infantile hemangioendothelioma. AB - During a routine physical exam, an 18-day-old male infant was noted to have persistent abdominal distention and prominent vascular markings over his abdominal wall. Laboratory studies were significant for an elevated alpha fetoprotein level of 7051 ng/ml and mild anemia. Abdominal ultrasound and CT scan demonstrated a large, heterogeneous mass in the lateral segment of the left lobe of the liver. Although the patient did not have congestive heart failure or coagulopathy, surgical resection was performed to rule out malignancy. Histopathologic examination revealed a type I infantile hemangioendothelioma. This case report reviews the presentation and treatment of infantile hemangioendotheliomas and the differential diagnosis of neonatal hepatic neoplasms. PMID- 11880744 TI - Fever without apparent source on clinical examination. PMID- 11880745 TI - Osteomyelitis in children. PMID- 11880746 TI - Lower respiratory infections in children. PMID- 11880747 TI - On revealing what we'd rather hide: the problem of describing study participation. PMID- 11880748 TI - Air pollution and Epidemiology: "deja vu all over again?". PMID- 11880749 TI - Where have all the winos gone? PMID- 11880750 TI - Reporting participation in case-control studies. AB - Although low participation in case-control studies may threaten their validity, it is difficult to evaluate the potential problem of nonresponse across studies because the level of participation among subjects is not reported in a consistent manner. This commentary presents a proposal for standard reporting of the process by which control subjects are recruited to a study and the outcomes of recruitment attempts. We encourage authors to provide a description of the sampling frame, the method of selecting units from the frame, and the number of units selected. For each unit selected, an outcome should be classified into four broad categories: ineligible sampling unit, unable to determine eligibility, respondent not eligible, and respondent screened and eligible. Details should be provided in each category; these details would differ according to the method used to contact potential respondents (by random-digit dialing, by telephone from a list, in person, or by mail). The proportion who participated can be reported as the investigator sees fit, as long as reporting of the components is complete. The sampling frame and outcomes should be reported separately for various parts of the study, such as controls from different sources or those who gave biological specimens. Standardized reporting will allow an evaluation of the success of various methods in recruiting potential participants, and will contribute important information for assessing the validity of individual studies. PMID- 11880751 TI - Alcoholic beverage preference and risk of becoming a heavy drinker. AB - BACKGROUND: Studies have suggested that wine drinkers are at lower risk of death than beer or spirits drinkers. The aim of this study is to examine whether the risk of becoming a heavy or excessive drinker differs among individuals who prefer different types of alcoholic beverages. METHODS: In a longitudinal study of 10,330 moderate drinkers from Copenhagen, Denmark, we used logistic regression analyses to address the risk of becoming a heavy or excessive drinker (above 14 and 21 drinks per week, respectively, for women and above 21 and 35 drinks per week for men) according to preference of wine, beer, or spirits. RESULTS: Compared with those who preferred wine, those who preferred beer tended to have increased risk of becoming heavy and excessive drinkers. Women who preferred beer had odds ratios of 1.14 (95% CI = 0.87-1.50) for becoming heavy drinkers and 1.50 (95% CI = 0.93-2.43) for becoming excessive drinkers. For men who preferred beer the ORs were 1.16 (95% CI = 0.84-1.58) and 1.81 (95% CI = 0.85-3.82). CONCLUSION: The finding that moderate wine drinkers appear to be at lower risk of becoming heavy and excessive drinkers may add to the explanation of the reported beverage specific differences in morbidity and mortality. PMID- 11880752 TI - Economic antecedents of mortality among the very old. AB - BACKGROUND: The 140-year trend of increased longevity in the developed world continues with no end in sight. Much of this trend has been attributed to mechanisms in which the benefits of economic growth act on the young. No attempts have been made to reconcile these theories with the experience of the very old. METHODS: Using Swedish data for the years 1861 through 1999, I examine the relationship between manufacturing wages and the odds of dying among persons 90 94 years of age. RESULTS: The findings imply that the odds of death among persons 90-94 years of age in Sweden for the years 1862 through 1999 declined about 0.1319 with every 1% increase in wages paid to mining and manufacturing workers. The median percentage increase in wages, 4.6%, therefore implies a decrease of 0.6% in the odds of mortality. CONCLUSIONS: The discovered effect supports the notion that increases in wealth have contributed to decreasing mortality among the very old over the last century and a half. The findings imply that theories focusing entirely on the young cannot fully account for increased life expectancy in populations with growing fractions of the elderly. PMID- 11880753 TI - Active and passive smoking in breast cancer: prospective results from the Nurses' Health Study. AB - BACKGROUND: The role of active and passive smoking in breast cancer remains controversial. METHODS: Using data collected in the prospective Nurses' Health Study, we examined the influence of active and passive smoking on the incidence of invasive breast cancer. The analysis was based on women responding to the 1982 questionnaire, which included questions on passive smoking exposure. Information on active smoking was collected in biennial questionnaires. A total of 78,206 women were followed prospectively from 1982 until June 1996. RESULTS: Of these women, 3,140 reported a diagnosis of invasive breast cancer during follow-up. Compared with never active smoking, relative risks (RR) of breast cancer were 1.04 (95% CI = 0.94-1.15) for current active smoking and 1.09 (95% CI = 1.00 1.18) for past active smoking. The RR for regular passive exposure at work and at home was 0.90 (95% CI = 0.67-1.22). For active smoking, a modest increase in risk was confined to women who began smoking before the age of 17 (RR = 1.19; 95% CI = 1.03-1.37). CONCLUSION: Results suggest that passive smoking is unrelated to breast cancer. However, results for active smoking are compatible with a small increase in risk when smoking is initiated at young ages. PMID- 11880754 TI - Leukemia in childhood and adolescence and exposure to ionizing radiation in homes built from uranium-containing alum shale concrete. AB - Concerns in Sweden about indoor radon around 1980 prompted measurements of gamma radiation from the facades of houses to identify those constructed of uranium containing alum shale concrete, with potentially high radon concentrations. To evaluate any possible risk of acute lymphocytic leukemia from exposure to elevated gamma-radiation in these homes, we identified the acute lymphocytic leukemia cases less than 20 years of age in Sweden during 1980-1989 as well as eight controls per case from the population registry, matching on age, gender, and county. Using the existing measurements, exposure was assessable for 312 cases and 1,418 controls from 151 properly measured municipalities. A conditional logistic odds ratio of 1.4 (95% confidence interval = 1.0-1.9) was obtained for those ever having lived in alum shale concrete houses, with the average exposure exceeding 0.10 microsieverts per hour. Comparing those who ever lived in alum shale concrete houses (divided by higher and lower annual average exposure) with those who never lived in such houses, we found a weak dose-response relation. The results suggest some risk of acute lymphocytic leukemia from indoor ionizing radiation among children and young adults. PMID- 11880755 TI - Adult leukemia: a spatial analysis. AB - A simple and direct analysis of the spatial distribution of adult leukemia data results from a geopolitical map transformed to have a uniform density of the population at risk. Geographic displays and statistical assessments then lead to a series of informative descriptions of the observed spatial pattern for six sex- and age-specific categories of non-Hispanic white individuals. Using a statistical/graphical approach, no consistent pattern of disease is observed; however, indications emerge of isolated nonrandom influences among young females (ages 10-29 years). PMID- 11880756 TI - Nonfatal cancer preceding Parkinson's disease: a case-control study. AB - BACKGROUND: We conducted a population-based case-control study to investigate the association of Parkinson's disease (PD) with preceding nonfatal cancer. METHODS: We used the medical records-linkage system of the Rochester Epidemiology Project to identify all incident cases of PD in Olmsted County, MN (1976-1995). Each case was matched by age and sex to a general population control. We ascertained cancer diagnoses through medical records abstraction. RESULTS: The frequency of any cancer was lower in cases (19.4%) than in controls (23.5%) (OR = 0.79; 95% CI = 0.49-1.27). This pattern was more pronounced in women than in men, and in patients age 71 years or younger at onset of PD than in older patients. We found an interaction between smoking and smoking-related cancers in their association with PD. Bladder cancer (OR = 0.22; 95% CI = 0.03-2.24) and breast cancer (OR = 0.20; 95% CI = 0.02-1.71) were less frequent in PD cases than in controls, whereas prostate cancer was more frequent in PD cases than in controls (OR = 1.80; 95% CI = 0.60-5.37). However, these results are based on small numbers. CONCLUSIONS: We did not find a strong association between PD and preceding nonfatal cancer. There were suggestive trends in analyses stratified by sex and age at onset of PD, and for specific cancers related to smoking or hormonal factors. PMID- 11880757 TI - Heterogeneity in assessing self-reports of caffeine exposure: implications for studies of health effects. AB - BACKGROUND: Coffee and its metabolite caffeine are widely studied for their health effects but with inconclusive results. Caffeine is particularly difficult to assess, and therefore we explore heterogeneity of caffeine exposure. METHODS: We categorized caffeine exposure among 2,478 pregnant women in southern New England during 1996-2000 by the traditional laboratory-based methods of M. Bunker and M. McWilliams. A subsample was examined to ascertain caffeine levels of brewed or purchased beverages actually consumed. RESULTS: More than half (56.6%) of women drank coffee since becoming pregnant. Serving sizes ranged from 2 to 32 oz and are considerably larger than laboratory standards, which are typically 8 10 oz, as compared with the standard of 5 to 6 oz. Conversely, caffeine content per serving of coffee was one-third the laboratory standard, eg, 100 mg caffeine compared with 300 mg for a 10-oz cup. Tea brewed more than 3 minutes contained 42 mg caffeine as compared with the standard of 94 mg. When the amount of caffeine actually consumed was measured, one-quarter (24.8%) of subjects traditionally classified as consuming 300+ gm caffeine daily were reclassified as consuming 150 299 mg. CONCLUSION: Misclassification of caffeine consumption increases difficulty in identifying health effects from caffeine. Some combination of more precise consumption data and a biomarker such as paraxanthine may more precisely estimate exposure. PMID- 11880758 TI - Air pollution and distributions of lymphocyte immunophenotypes in cord and maternal blood at delivery. AB - A cross-sectional study of deliveries in two districts in the Czech Republic, 1994-1996, assessed the relation between air pollution and lymphocyte immunophenotype distributions. Maternal and cord blood samples were assayed by flow cytometry within 24 hours of delivery for 303 deliveries from Teplice, a polluted district, and 215 from Prachatice, a less polluted district. Analyses focused on: CD3(+) T-lymphocytes, CD3(-) CD19(+) B-lymphocytes, and CD3(-) CD16(+)56(+) natural killer (NK) lymphocytes, as well as the subsets CD3(+)CD4(+) ("T-helper") and CD3(+)CD8(+) ("T cytotoxic/suppressor") and the ratio of these two lymphocytes. We collected reproductive, occupational, and life-style information by questionnaire, and abstracted data on labor and delivery from medical records. After adjustment for numerous risk factors in multivariate linear regression models fit for each lymphocyte subset, mothers from Teplice had lower percentages of total T-cells and of CD4(+) cells, and a lower ratio of CD4(+):CD8(+) cells. Cord bloods from Teplice had a higher percentage of NK cells and a less precise lower percentage of T-cells. Stronger differences in maternal lymphocytes were seen when analyses were limited to the central hospital in each district. Heavy air pollution may affect the immune system in pregnant women and/or fetuses, reflecting an acute and/or chronic effect, although unmeasured confounders could also play a role. PMID- 11880759 TI - Physical activity, body mass index, and ovulatory disorder infertility. AB - Few studies have examined whether activity and adiposity levels typical of American women affect their risk of ovulatory disorder infertility, and none has examined moderate and vigorous intensity exercise separately. We investigated these associations in the Nurses' Health Study II, comparing prospectively collected data on adiposity and activity for 830 cases of incident ovulatory infertility and 26,125 pregnancies. We observed a U-shaped association between body mass index (BMI) and relative risk of ovulatory infertility, with increased risk for BMI below 20.0 or above 24.0 kg/m2. On the basis of the BMI distribution of U.S. women, these findings suggest that 12% (95% confidence interval = 7-20%) of ovulatory infertility in the U.S. may be attributable to underweight (BMI <20.0) and 25% (95% CI = 20-31%) to overweight (BMI > or = 25.0). An increase in vigorous activity (but not moderate activity) was associated with reduced relative risk of ovulatory infertility. Each hour per week of vigorous activity was associated with a 7% (95% CI = 4-10%) lower relative risk of ovulatory infertility. After adjustment for BMI, a 5% (95% CI = 2-8%) reduction in relative risk per hour of weekly activity remained. These data suggest that, among American women, more ovulatory infertility is attributable to overweight and a sedentary lifestyle than to underweight and overexertion. PMID- 11880760 TI - Prolonged time to pregnancy in female workers exposed to ethylene glycol ethers in semiconductor manufacturing. AB - BACKGROUND: Previous research on reproductive effects of working in the semiconductor industry is limited and has produced conflicting results. METHODS: This retrospective cohort study was designed to investigate the risk factors for female fertility in a wafer-manufacturing company of Taiwan in 1997. Waiting time to pregnancy and potential confounders were collected by face-to-face interview. Exposure was assessed by directors and senior engineers in manufacturing, administrative, and safety and health departments according to safety records, personnel records, and job histories from questionnaires, using a tiered exposure assessment approach. Of a total 842 female workers, 720 (85.5%) participated. There were 292 pregnancies from 173 workers eligible for analysis. Relative fecundability (fecundability ratio, or FR) was calculated using Cox's proportional hazard models to evaluate the effect of occupational exposure to ethylene glycol ethers on female fertility. RESULTS: Waiting time to pregnancy of female workers in the photolithography area was longer than that of those in the nonfabrication area (FR = 0.77; 95% CI = 0.45-1.32), and those who were potentially exposed to ethylene glycol ethers showed longer time to pregnancy compared with those not exposed (FR = 0.59; 95% CI = 0.37-0.94). CONCLUSION: This study provides further evidence that ethylene glycol ethers may cause female subfertility. PMID- 11880761 TI - Maternal, delivery, and perinatal characteristics associated with cryptorchidism: a population-based case-control study among births in Washington State. AB - BACKGROUND: The etiology of cryptorchidism is largely unknown. To identify maternal, perinatal, and delivery characteristics associated with cryptorchidism at birth, we conducted a population-based case-control study using Washington State birth certificates linked to birth hospitalization records. METHODS: We identified 2,395 cases of cryptorchidism among male infants born in Washington State during 1986-1996, and, for comparison, we randomly selected four controls per case (N = 9,580), frequency-matched by year of birth. RESULTS: Infant characteristics associated with cryptorchidism included low birth weight (OR = 1.5; 95% CI = 1.3-1.8), small size for gestational age (OR = 1.9; 95% CI = 1.6 2.2), and breech presentation (OR = 1.7; 95% CI = 1.4-2.1). In addition to cryptorchidism, cases were more likely to have another type of congenital malformation (OR = 3.7; 95% CI = 3.2-4.2), particularly digestive (OR = 6.8; 95% CI = 3.7-12.7) or genitourinary (OR = 4.1; 95% CI = 3.0-5.6). Maternal and pregnancy characteristics associated with cryptorchidism included nulliparity (OR = 1.2; 95% CI = 1.1-1.3), maternal smoking during pregnancy (OR = 1.2; 95% CI = 1.1-1.4), and the following pregnancy complications: oligohydramnios (OR = 1.8; 95% CI = 1.3-2.6), placental abnormality (OR = 1.3; 95% CI = 1.0-1.8), and pregnancy-induced hypertension (OR = 1.6; 95% CI = 1.4-1.9). Odds ratios were similar when the analysis was restricted to term infants. CONCLUSIONS: These findings suggest that factors affecting fetal growth and development may increase the risk of cryptorchidism. PMID- 11880762 TI - Growth in girls exposed in utero and postnatally to polybrominated biphenyls and polychlorinated biphenyls. AB - BACKGROUND: Accidental contamination with polybrominated biphenyls (PBBs) of the Michigan food supply in 1973 led to the exposure of more than 4000 individuals and to formation of the PBB cohort registry (1976-1979). At enrollment, measurements were taken of serum PBB and polychlorinated biphenyl (PCB), possible endocrine disrupting chemicals. METHODS: We examined the association of estimated PBB and PCB exposure during pregnancy with current height and weight in 308 daughters, 5-24 years of age (mean age 15.2 years), born to women in the cohort. We estimated prenatal PBB exposure using maternal enrollment serum PBB and a model of PBB elimination. Prenatal PCB exposure was estimated using maternal enrollment serum PCB because background-level exposure through diet was ongoing. Self-reported height and weight were obtained from a 1997-1998 health survey. RESULTS: We found no association between prenatal PBB exposure and either daughter's current height or daughter's weight adjusted for height; however, prenatal PCB exposure above 5 parts per billion was associated with reduced weight adjusted for height. Exposure through breastfeeding did not modify the association. CONCLUSIONS: Mothers with PCB levels above the median had daughters whose current weights were 11 pounds lower than that of the daughters whose mothers had levels below the median. This study provides evidence that prenatal exposure to PCBs may affect growth. PMID- 11880763 TI - Seasonal variation in hospital discharge diagnosis of atrial fibrillation: a population-based study. AB - BACKGROUND: Cardiovascular events such as acute myocardial infarction and stroke vary seasonally, with a peak incidence during the winter and a trough during the summer. Few data exist on the seasonal variation of atrial fibrillation. METHODS: We examined the association among season, outdoor temperature, and risk of atrial fibrillation. We identified 32,992 subjects, age 50-89 years, in a random sample of half the people with an incident hospital discharge diagnosis of atrial fibrillation in the Danish National Hospital Discharge Register from 1980 to 1993. We used a periodic regression model to evaluate any seasonal distribution. RESULTS: We found a winter peak and a summer trough in the incidence of atrial fibrillation. The relative incidence of atrial fibrillation during winter as compared with summer, estimated as the ratio of the incidence in the month of the peak relative to the incidence in the month of the trough, was small (relative risk = 1.20; 95% confidence interval = 1.12-1.29). Stratification by sex, age, and the presence or absence of diabetes and underlying cardiovascular diseases did not affect the seasonal pattern. We also found an inverse relation between mean outdoor temperature and incidence of atrial fibrillation. CONCLUSION: The risk of atrial fibrillation was modestly higher during the winter and was inversely associated with outdoor temperature. PMID- 11880764 TI - Dietary and adipose tissue gamma-tocopherol and risk of myocardial infarction. AB - BACKGROUND: Gamma-tocopherol, the most abundant form of dietary vitamin E, may lower the risk of coronary heart disease. METHODS: We investigated whether dietary and adipose tissue gamma-tocopherol are associated with myocardial infarction (MI) in 475 survivors of a first MI and 479 controls from a population based study carried out between 1994 and 1998 in Costa Rica. Dietary intake was assessed with a validated food-frequency questionnaire and an adipose tissue sample. Conditional logistic regression was used to calculate odds ratios (OR) and 95% confidence intervals (CI). RESULTS: Subjects in the highest quintile of dietary gamma-tocopherol had a lower risk of MI compared with those in the lowest quintile (OR = 0.76; 95% CI = 0.50-1.17, P = 0.02 for trend). This trend was no longer statistically substantial in multivariate analysis (P = 0.44). A weak association was found for adipose tissue gamma-tocopherol in univariate (OR = 1.46; 95% CI = 0.94-2.27) and multivariate (OR = 1.31; 95% CI = 0.62-2.76) models. A substantial inverse association with MI was found for total dietary vitamin E whether supplement users were included or excluded (P = 0.01 and 0.05 for trend, respectively). CONCLUSION: These data suggest that gamma-tocopherol does not protect against nonfatal MI. PMID- 11880765 TI - Measuring behavioral interventions by questionnaires and prospective diaries: an example of sunscreen use. AB - BACKGROUND: Diaries are used to measure behavior in intervention trials, despite the tendency of diaries to overestimate adherence. METHODS: We estimated the frequency of sunscreen application using prospective diaries and a questionnaire in a trial evaluating sunscreen use. RESULTS: Sunscreen application was reported more frequently in the diaries than in the questionnaire by the intervention group. This was supported by an increase in the weight of sunscreen used during the times participants were keeping diaries. CONCLUSIONS: Diaries may be better as an aid to increase adherence than as a tool for its estimation. Use of diaries did not overestimate the frequency of application in the control group. PMID- 11880766 TI - The protective effect of alcoholic beverages on the occurrence of a Salmonella food-borne outbreak. AB - BACKGROUND: There are few studies about the effect of alcoholic beverages in food borne disease outbreaks. METHODS: We studied this effect during an outbreak of Salmonella ohio infection that occurred in Castellon (Spain) in May 2000. S. ohio was isolated in stools of patients and also in implicated foods. RESULTS: Analysis was restricted to the group that was exposed to contaminated food (51 persons). We compared attack rates by level of alcohol intake (none, 1-40 gm, and >40 gm). The adjusted relative risks in the two alcohol categories were 0.73 [95% confidence interval (CI) = 0.46-1.15] and 0.54 (95% CI = 0.31-0.96), respectively. CONCLUSIONS: We discuss the possible protective effect of alcoholic beverages in some food-borne infectious outbreaks. PMID- 11880768 TI - Noxious exposures in leisure time and risk of lung cancer: a neglected exposure? PMID- 11880767 TI - Race, epithelial ovarian cancer survival, and membership in a large health maintenance organization. AB - BACKGROUND: African-American ovarian cancer patients present with more advanced disease and have poorer survival than do white patients. METHODS: To determine whether these differences occur among African-American and white patients who have equal access to medical care, we analyzed ovarian cancer patient characteristics separately for 1,587 members of the Kaiser Permanente Medical Plan of Northern California and 5,757 non-members. RESULTS: The distributions of disease stage at diagnosis were similar among African-American and white patients, both in the Kaiser plan and elsewhere. However, ovarian cancer death rates, adjusted for disease stage and age at diagnosis and for histology, were higher for African-American patients compared with white patients, regardless of Kaiser membership status. The death rate ratios for African-Americans compared with whites were 1.32 (95% CI = 1.02-1.70) for Kaiser members and 1.20 (95% CI = 1.04-1.40) for Kaiser non-members. CONCLUSION: Further research within an equal access care system is needed to evaluate other important factors such as specialty of surgeon, extent of residual tumor after surgery, chemotherapy treatment, and postoperative management to determine whether these factors are contributing to the differences in survival between African-American and white ovarian cancer patients. PMID- 11880769 TI - Is the effect of wine on health confounded by diet? PMID- 11880770 TI - Magnetic fields and miscarriage. PMID- 11880771 TI - Increased red cell folate concentrations in women of reproductive age after Canadian folic acid food fortification. PMID- 11880772 TI - Use of systemic corticosteroids and risk of esophageal cancer. PMID- 11880774 TI - BEWARE OF CERTAINTY WHERE NONE EXISTS: A REVIEW OF. PMID- 11880773 TI - Toxicity of fine particles. PMID- 11880775 TI - Use of an injectable calcium phosphate bone cement in the treatment of tibial plateau fractures: a prospective study of twenty-six cases with twenty-month mean follow-up. AB - OBJECTIVE: To evaluate the potential benefit of a new injectable mineral bone cement (Norian SRS, Cupertino, CA, USA) for the treatment of tibial plateau fractures OTA types 41.B2-B3 and C3. DESIGN: Prospective study with established protocol. SETTING: University hospital; university teaching hospital. PATIENTS: Twenty-six patients gave informed consent to participate in this study and were available for follow-up examinations. All patients had sustained intraarticular tibial plateau fracture types OTA B2-B3 and C3. All cases were followed with a mean follow-up time of 19.7 months (6 to 36 months, median eighteen months). INTERVENTION: Twenty-five cases were treated with open reduction, osteosynthesis with screws or plate, and injection of Norian SRS in the subchondral bone defect. One case had closed reduction, screw osteosynthesis, and percutaneous cement injection. MAIN OUTCOME MEASUREMENTS: Radiographs in two planes were evaluated prospectively at six, twelve, and twenty-six weeks postoperatively and at last follow-up using Rasmussen's radiologic score. Clinical parameters were measured at the same time intervals using Lysholm's and Tegner's knee scores. RESULTS: Two cases required early wound revisions because of sterile drainage. Two cases developed partial loss of reduction of the fracture between four and eight weeks after surgery, requiring revision surgery in one case (total complication rate 15.3 percent). No other patients had complications, and all other fractures healed without any displacement. The high mechanical strength of the cement allowed early weight bearing after a mean postoperative period of 4.5 weeks (1 to 6 weeks). CONCLUSIONS: An injectable mineral bone cement with high initial mechanical strength was used to fill bone defects in unstable tibial plateau fractures with good success. This material offers new perspectives in the treatment of tibial plateau fractures. PMID- 11880776 TI - The use of the unreamed AO femoral intramedullary nail with spiral blade in nonpathologic fractures of the femur: experiences with eighty consecutive cases. AB - OBJECTIVE: To evaluate the unreamed femoral nail with spiral blade (UFN-SB) in the treatment of nonpathologic subtrochanteric and segmental femoral fractures. DESIGN: Prospective follow-up of eighty consecutive fractures in eighty patients still alive at least ten months after the operation. SETTING: Academic teaching hospital. PATIENTS: Eighty patients with a subtrochanteric or segmental femoral fracture. Excluded were patients with pathologic fractures and those who died within ten months of the operation. INTERVENTION: Unreamed femoral nail with spiral blade. MAIN OUTCOME MEASURES: Clinical and radiographic examination. RESULTS: In seventeen fractures, the UFN-SB failed before bony union (21 percent): bending of the spiral blade, five times; migration, nine times; and breakage, three times. Revision surgery was necessary in seven cases (9 percent). All complications except one were observed in elderly women with a Seinsheimer fracture type IIC or V. Nevertheless, all fractures healed within one year, including those that needed revision surgery. CONCLUSIONS: There is predictive value of the Seinsheimer classification as to outcome using UFN-SB. The UFN-SB is an option for the treatment of subtrochanteric or segmental fractures of the femur, especially in patients with a good quality of bone. The complications using this device are not caused by the learning curve, but by the characteristics of the implant and the type of fracture for which it is used. The implant should not be used in elderly women with a reversed oblique fracture or a subtrochanteric fracture with an intertrochanteric component. PMID- 11880777 TI - Histologic characterization of acute spinal cord injury treated with intravenous methylprednisolone. AB - OBJECTIVE: Many substances have been investigated for attenuation of spinal cord injury after acute trauma; however, pharmacologically only steroid administration has shown clinical benefits. This study attempts to characterize local spinal cord histologic response to human dose equivalent (HDE) intravenous methylprednisolone (MP) administration in a rodent model of acute spinal cord injury. DESIGN: Forty-eight Sprague-Dawley rats were divided equally into control and experimental groups. Each group was subdivided into eight sets of three animals each, according to postinjury intervals. Paraplegia after lower thoracic laminectomy was achieved using a standardized weight drop technique. INTERVENTION: Within one hour, experimental animals were treated with HDE MP followed by 23-hour continuous infusion of HDE MP. Spinal cords were harvested at variable intervals postinjury and prepared for histologic/immunohistochemistry examination. MAIN OUTCOME MEASUREMENTS: Edema, necrosis, and glial fibrillary acidic protein (GFAP) positivity in the specimens from treated/control groups were graded by microscopy and immunohistochemistry staining and compared in a blinded manner by a qualified neuropathologist and senior authors. RESULTS: Minimal differences were observed between control and MP-treated animals at zero and four hours. At eight hours, increased white matter and medullary edema was evident in control versus MP-treated rats. This trend continued through twelve, sixteen, twenty-four, forty-eight, and seventy-two hours. No difference was observed in the astrocytic response to injury by GFAP immunohistochemistry between the groups. CONCLUSIONS: Histologically, MP reduces the development of severe edema and preserves spinal cord architecture adjacent to the site of injury. In contrast, MP does not alter the development of spinal cord necrosis or astrocytic response at the zone of injury. PMID- 11880778 TI - High-pressure pulsatile lavage irrigation of fresh intraarticular fractures: effectiveness at removing particulate matter from bone. AB - OBJECTIVE: To determine the effectiveness of high-pressure pulsatile lavage (HPL) versus bulb syringe (BS) irrigation in removing particulate matter from metaphyseal cancellous bone. DESIGN: Four grams of particulate graphite were placed in twenty distal femoral intraarticular osteotomies performed on New Zealand rabbit hind limbs. Two groups of ten specimens were then irrigated using either HPL or BS irrigation. A representative coronal section from each specimen was then prepared for histologic evaluation using 400x light microscopy. The number and distribution of graphite particles-present as small (less than 20 micrometers), medium (20 to 50 micrometers), and large (greater than 50 micrometers) aggregates-were then recorded. RESULTS: The mean maximum perpendicular distance of graphite aggregates of all sizes from the osteotomy site was 12.4 millimeters (+/-SD 2.5) in the HPL group and 12.5 millimeters (+/ SD 2.0) in the BS group (p > 0.5). The mean number of aggregates within four 400x fields (1.08 millimeters) of the osteotomy site was 21.9 (+/-SD 22.0) in the HPL group and 21.8 (+/-SD 27.5) in the BS group (p > 0.5). The mean total number of aggregates in the area surveyed was 129.4 (+/-SD 79.6) in the HPL group and 137.5 (+/-SD 113.6) in the BS group (p > 0.5). Separate analyses controlling for aggregate size of the specimens also revealed no significant differences between HPL and BS irrigation. CONCLUSION: HPL and BS irrigation appear equally effective in removing particulate matter from metaphyseal cancellous bone in an intraarticular fracture model. Furthermore, HPL does not appear to drive particulate matter farther into metaphyseal cancellous bone than BS irrigation. PMID- 11880779 TI - Minimal versus maximal plate fixation techniques of the ulna: the biomechanical effect of number of screws and plate length. AB - OBJECTIVE: To test the hypothesis that longer plates with the minimum number of screws provide equivalent or superior strength of fixation to standard compression plating using the maximum number of screws. DESIGN: Prospective analysis of biomechanical data was performed. SETTING: In vitro experimentation. INTERVENTIONS: A reproducible osteotomy was made in formalin-fixed ulnae. The osteotomies were stabilized employing six-, eight-, or ten-hole plates with two screws in the outermost holes and two screws in the innermost holes, and compared to stabilization with a six-hole plate with six screws. MAIN OUTCOME MEASURE: Four-point mechanical testing to failure was performed in both apex-dorsal (tension-band) or medial-lateral bending modes. Load-displacement curves were obtained. RESULTS: All of the longer plates with the minimum number of screws were stronger than the six-hole plate with six screws when tested in the medial lateral and the tension-band mode. The eight-hole plate with four screws, however, was statistically inseparable from the fully loaded six-hole plate. There was no statistical difference between any of the configurations in regard to the stiffness of the fixation. CONCLUSIONS: In a cadaveric ulnar osteotomy model stripped of soft tissue, the number of screws is less important than the length of the plate in providing bending strength to the construct. PMID- 11880780 TI - Split depression tibial plateau fractures: a biomechanical study. AB - OBJECTIVE: To determine the biomechanical characteristics of four different fixation constructs for split depression fractures of the lateral tibial plateau (OTA classification 41B3.1). DESIGN: Laboratory investigation using a cadaveric simulated split depression tibial plateau fracture model. SETTING: Split depression tibial plateau fractures were created, reduced, and instrumented in a matched pair design. Specimens were tested for stiffness using a materials testing machine. INTERVENTION: Tibias were instrumented with an L-buttress plate, four 3.5-millimeter subchondral raft screws with an antiglide plate, an L buttress plate with cancellous allograft, or four 3.5-millimeter subchondral raft screws placed through a periarticular plate. MAIN OUTCOME MEASUREMENTS: Vertical subsidence of the lateral tibial plateau was measured for the entire construct and for the local depression. The relative medial and lateral condylar tilt with central loading was also measured. RESULTS: There was no significant difference between the four fixation methods for overall longitudinal stiffness of the proximal tibial fracture fixation construct (range, 2,026 to 2,666 newtons per millimeter). The local depression stiffness for the raft-periarticular plate and raft-antiglide plate were 425 newtons per millimeter and 342 newtons per millimeter, respectively, versus 243 newtons per millimeter and 210 newtons per millimeter for the two large fragment buttress constructs. There was no significant difference between the local depression stiffness for the two raft constructs. There was no significant difference between the local depression stiffness for the two buttress plate constructs. Local depression stiffness was found to be significantly greater for the raft-periarticular plate construct when compared with the large fragment buttress plate construct without bone graft (p = 0.0314). Condylar tilt data showed a significant difference between the medial tilt observed in the prefracture specimen and the lateral tilt observed after fixation (p less-than-or-equal 0.017) for all constructs. CONCLUSIONS: There was no significant difference in the overall construct stiffness between the four fixation constructs. Fixation constructs with a raft of subchondral screws were more resistant to local depression loads. This supports the use of a raft construct when a central depression is a significant component of the overall fracture pattern. Condylar tilt data showed a persistent weakness in the postfixation lateral plateau regardless of fixation construct when compared with the intact specimen. This supports the current clinical practice of delayed weight-bearing for ten to twelve weeks. PMID- 11880781 TI - Screw pullout strength: a biomechanical comparison of large-fragment and small fragment fixation in the tibial plateau. AB - OBJECTIVES: To compare the pullout strengths of 6.5-millimeter diameter partially threaded cancellous screws and 4.5-millimeter diameter fully-threaded cortical screws versus 3.5-millimeter diameter cortical screws in the proximal tibia. DESIGN: Three screws were inserted in the lateral tibial plateau of each leg of fifteen paired cadaveric tibias. In one tibia, large-fragment fixation was used, consisting of a unicortical 6.5-millimeter screw in the subchondral bone, and bicortical 4.5-millimeter screws in the metadiaphyseal and diaphyseal bone. In the contralateral tibia, small-fragment fixation consisting of three 3.5 millimeter screws was used, placing the screws in the same positions as described above. MAIN OUTCOME MEASUREMENTS: A materials-testing machine was used to determine axial pullout strengths of each screw. The mean pullout strengths of large-fragment and small-fragment screws in each position were compared. RESULTS: No significant difference in pullout strengths was found between the large fragment and small-fragment screws in subchondral and metadiaphyseal bone. A statistically significant difference was found between pullout strengths of large fragment and small-fragment screws in diaphyseal bone. CONCLUSIONS: In human proximal tibial bone, the data from this study do not suggest that the pullout strength of 3.5-millimeter screws differs from that of 6.5-millimeter screws in subchondral bone, or that the pullout strength of 3.5-millimeter screws differs from that of 4.5-millimeter screws in metadiaphyseal bone. However, the pullout strength of 3.5-millimeter screws is significantly less than that of 4.5 millimeter screws in diaphyseal bone. The authors of the present study believe this supports the use of small-fragment fixation in the treatment of tibial plateau fractures. PMID- 11880782 TI - A method for delivering variable impact stresses to the articular cartilage of rabbit knees. AB - OBJECTIVE: To develop a method by which a single impact force of controlled magnitude and rate could be applied uniformly to an area on the posterior aspect of the medial femoral condyle of adult rabbits. DESIGN: An in-vivo animal model using a pendulum device, designed and manufactured to supply the kinetic energy necessary to apply different impact loads to the posterior aspect of the medial femoral condyle of a rabbit. SETTING: Biomechanical laboratory, University Medical Center. SUBJECTS: A total of thirty-six femoral condyles from 3-kilogram New Zealand White (NZW) rabbits were used during this evaluation. INTERVENTION: An aluminum impactor was made based on the sagittal and coronal radii of curvature of six matched pairs (n = 12) of femurs from three-kilogram NZW rabbits. This impactor was coupled with the pendulum and used to apply different impact loads to both of the medial femoral condyle of the knees of NZW rabbits (n = 24). MAIN OUTCOME MEASUREMENTS: Peak impact force, time to peak impact force, and average contact area between impactor and medial femoral condyle, were measured for each group of animals tested. RESULTS: The pendulum delivered a consistent impact force to the rabbit condyle of 120.0 N (+/-18.1; coefficient of variance, 15 percent) with 400 grams attached to the pendulum arm, at an average time to peak force of 0.021 seconds (+/-0.001, co. var. 4.8 percent). The peak impact force was significantly different for each of the three impact mass groups of animals (p < 0.001). By contrast, time to peak force for each mass group averaged approximately 0.020 seconds and the average contact area was 6.26 mm2 (+/-0.51). Quantitative assessment of the exposed medium pressure-sensitive film confirmed uniform impact force intensity within each specimen. CONCLUSIONS: An in vivo animal model was developed to deliver a controlled and rapid impact force to a specific area of the weight-bearing surface of the adult rabbit knee. These loads were applied at a rate comparable to the clinical setting of falling onto an outstretched hand, thus simulating a common clinical scenario by which cartilage is often injured. This model can be used in future experiments to investigate mechanism by which posttraumatic arthritis develops after articular injuries. PMID- 11880784 TI - The centered lateral fluoroscopic image of the knee: the key to safe tensioned wire placement in periarticular fractures of the proximal tibia. AB - The importance of using the centered lateral view of the knee for placement of tensioned wires during the treatment of periarticular fractures of the proximal tibia is illustrated. The technique of using a horizontal reference wire as a landmark for subsequent wire placement is described for circular fixators and hybrid fixators. PMID- 11880783 TI - State of the art review: techniques to avoid pin loosening and infection in external fixation. AB - The purpose of this manuscript is to review techniques of optimizing the interface between the bone and pin in external fixation to minimize pin loosening and infection. CONCLUSIONS: Among the different techniques to improve the bone pin interface in external fixation, coating the pins with hydroxyapatite proved to be the most effective. In a highly loaded animal study, three pin types were compared. Type A remained uncoated, type B was coated with hydroxyapatite, and Type C was coated with titanium. Radiographic rarefaction of the bone pin tract was lower in type B pins. Extraction torque was thirteen times higher in type B pins compared to type A and two times higher compared to type C pins. Extraction torque was significantly lower compared to the corresponding insertion torque in both types A and C. In contrast, in the hydroxyapatite coated pins there was no difference between extraction and insertion torque. At sixty times magnification, bone pin contact of type B and C pins was significantly higher than type A. At 10,000 times magnification direct bone pin contact was found only in type B pins. In a clinical study the pin insertion and extraction torque forces were measured in a study of seventy-six external fixation pins in nineteen patients treated with hemicallotasis for osteoarthritis of the medial side of the knee. The patients were randomized to be treated with either standard tapered pins or tapered pins coated with hydroxyapatite. Extraction torque of the hydroxyapatite coated pins was higher than the standard ones in both cancellous and cortical bone. These studies show that in hydroxyapatite coated pins there is no deterioration of the bone-pin interface strength and there is optimal bone-pin contact. Among the various pin types coated with hydroxyapatite, the best results were obtained with the tapered pins. PMID- 11880785 TI - Vascular compromise after insertion of a retrograde femoral nail: case report and review of the literature. AB - The authors describe entrapment of the popliteal artery within fragments of an open supracondylar femur fracture after reduction and stabilization with a retrograde femoral nail. At the time of fracture fixation, the loss of pedal pulses was noted, and an exploration of the popliteal vessels was performed. The popliteal artery was easily identified and dissected free after extension of the patient's lateral thigh wound. Following thrombectomy, blood flow to the distal extremity was restored and no postoperative vascular sequelae were encountered. PMID- 11880786 TI - Displacement of the common peroneal nerve associated with upper tibial fracture: implications for fine wire fixation. AB - The constant anatomic position of the common peroneal nerve is relied on when performing fine wire external fixation in the upper tibia. We report the case of a sixty-two-year-old woman with a Schatzker Type V fracture of her right tibial plateau and upper-third diaphyseal fracture associated with displacement and shortening of the upper tibia. She was treated by minimal internal fixation of the intraarticular fracture and application of a Sheffield Hybrid External Fixator. During percutaneous insertion of the reference wire in the fibular head, a distal muscle twitch alerted the surgeon, and the common peroneal nerve was duly explored and found displaced forward over the fibular head, dangerously close to the wire. It is postulated that at the time of injury, the common peroneal nerve was displaced anteriorly and that despite reduction of the tibial fractures, it had failed to return to its original position. The mechanism of this was confirmed by an anatomic study on an above-the-knee amputation specimen in which the metaphyseal-diaphyseal element of the fracture was reproduced. We recommend insertion of the reference fibular wire with the knee in flexion. Open insertion of this wire, with an incision down to bone and exposure of the fibular head, is recommended in cases in which severe trauma with shortening of the upper tibia, with possible disruption of the tibiofibular joint, puts the nerve in danger of injury. PMID- 11880787 TI - Traumatic divergent dislocation of the extensor tendons over the metacarpophalangeal joint. AB - Dislocation of the extensor tendon over the metacarpophalangeal joint (MCP) is rare. Involvement of the fifth ray is even more exceptional. The authors of the present study describe such a case that occurred in a professional musician. We report a divergent dislocation of both extensor tendons over the metacarpophalangeal joint of the fifth finger. PMID- 11880788 TI - Age, gender, work capacity, and worker's compensation in "patients with displaced intraarticular calcaneal fractures". PMID- 11880789 TI - Paradoxical fat embolism after intramedullary rodding. PMID- 11880790 TI - Do variations in breast cancer care matter? PMID- 11880791 TI - Persistent differences in sociodemographic determinants of breast conserving treatment despite overall increased adoption. AB - BACKGROUND: Use of breast-conserving treatment (BCT) has previously demonstrated variability by sociodemographic factors. OBJECTIVE: To determine whether variation in use of BCT by age, race, county income, county education, and population density declined between 1983 and 1996. DESIGN: Trends in use of BCT over time were modeled with logistic regression. SETTING: Surveillance, Epidemiology, and End Results national tumor registry data. PATIENTS: Population based cohort of 158,496 women with local or regional stage breast cancer. MAIN OUTCOME MEASURE: Receipt of BCT. RESULTS: Use of BCT increased overall, and among all subgroups of age, county income, county education, population density, and race. There was no decline in age-related variation in use of BCT over time. However, older women were less likely to undergo BCT including radiotherapy (RT) and lymph node dissection (LND), and were more likely to undergo BCT omitting RT and/or LND. Variation in use of BCT by county income persisted, with women residing in poorer counties less likely to undergo BCT, whether accompanied by RT and LND. Variation in overall use of BCT by county education also persisted. Although women residing in better-educated counties were more likely to undergo BCT accompanied by RT and LND, they were not more likely to undergo BCT omitting RT, LND, or both. No decline in variation by population density occurred, with women residing in urban areas more likely to use BCT whether accompanied by RT and LND. CONCLUSIONS: Sociodemographic differences in BCT use have persisted over time. The increased overall adoption of BCT has not led to consistency in use of this treatment. PMID- 11880792 TI - Using telephone interviews to reduce nonresponse bias to mail surveys of health plan members. AB - OBJECTIVE: To assess the nonresponse bias associated with mail-survey returns and the potential for telephone interviews with nonrespondents to reduce that bias. METHODS: A mail survey about health care experiences was conducted with samples of 800 members in each of four health plans. Subsequent attempts were made to interview nonrespondents by telephone. RESULTS: Response rates for the mail surveys averaged 46%; the telephone effort raised the average to 66%. On 17 of 19 measures of health status or need and use of health services, mail respondents were in poorer health and needed more services than interviewed nonrespondents. Thirteen of 36 reports and ratings of health care also differed significantly between the two groups. Based on administrative data, telephone interviews of mail nonrespondents improved the demographic representativeness of the responding samples. Adjusting mail returns to sample population characteristics could not replicate the dual-mode results. CONCLUSIONS: Returns to mail surveys are likely to be related to survey content and hence are potentially biased. Nonresponse to phone surveys is less directly related to survey content. Telephone interviews with mail nonrespondents not only increase response rates but also can produce less biased samples than mail-only protocols. PMID- 11880793 TI - Construct validity of medicare chemotherapy claims: the case of 5FU. AB - BACKGROUND: The elderly are under represented in clinical trials of cancer therapy and the elderly who are enrolled may be unrepresentative. OBJECTIVE: To assess whether Medicare claims data might be used to understand the benefits and tolerance of chemotherapy in the general elderly population, the construct validity of Medicare 5FU claims for elderly colon cancer patients within the SEER Medicare data set was determined. METHODS: In this validation study of Medicare chemotherapy claims from the linked the SEER-Medicare data set, the patterns of 5FU chemotherapy claims were evaluated for an incident cohort of elderly colon cancer patients (n = 15,039) during the 13 months following their diagnosis. Patterns of Medicare National Claims History (NCH) 5FU claims were evaluated with respect to prespecified patient-level disease and demographic factors from the data set. RESULTS: Twenty-two percent of patients had at least one detectable 5FU claim during the observation period. Among those patients, the median dose of 5FU was 1000 mg, the median interval between 5FU claims was 7 days, and the median number of claims during this period was 24. Multivariate regression revealed expected associations between demographic and disease factors and the likelihood of having a Medicare NCH 5FU claim. With increasing cancer stage, patients' likelihood of having a 5FU claim increased. Younger patients, married patients, white patients, patients with low comorbidity, and patients living in urban and less impoverished regions were each more likely to have 5FU claims. CONCLUSION: Because their pattern is consistent with the standard of medical care and with previously described associations with disease and demographic factors, it was concluded that Medicare NCH claims for 5FU administration in the SEER-Medicare data set exhibit construct validity. Criterion validation studies with an external gold standard should be pursued to determine the sensitivity and specificity of chemotherapy codes in the Medicare NCH files. PMID- 11880794 TI - Is health plan employer data and information set performance associated with withdrawal from medicare managed care? AB - BACKGROUND: Withdrawals of health plans from Medicare have affected more than 1.6 million beneficiaries. Some plans claim that providing higher quality care raises costs, lowers profits, and spurs withdrawal because plans cannot sustain high quality care under current payment levels. OBJECTIVE: To assess whether higher performance by Medicare health plans on quality indicators was associated with withdrawal. DESIGN: Retrospective cohort study. SUBJECTS: Taking each county where a contract was active as a unit of analysis, Medicare managed care plans active in 2310 contract-county combinations in 1997 were studied and followed for 3 years. MEASURES: Independent variables were scores on six indicators from the Health Plan Employer Data and Information Set (HEDIS) for each contract, collapsed into two summary measures: clinical and ambulatory care access. Separate Cox proportional hazards regressions were used for each indicator, and each summary measure, to assess the association of HEDIS performance with our outcome measure, time-to-withdrawal from Medicare. Multiple potential confounders were adjusted for. RESULTS: Of 2310 managed care contract-county combinations, 877 (38%) withdrew. The proportion of contract-counties with high scores on the summary clinical quality measure that withdrew was one-fifth that for low scorers (4.2% vs. 20.5%). For summary ambulatory care access performance, the corresponding ratio was two-fifths (12.8% vs. 32.0%). Lower payments were associated with higher withdrawal risk, but also higher clinical and ambulatory care access quality performance. In separate multivariable analyses controlling for confounders, both high clinical performance (HR, 0.18; 95% CI, 0.08-0.42) and high ambulatory care access performance (HR, 0.53; 95% CI, 0.27-1.07) were independently associated with lower withdrawal risk. CONCLUSIONS: Health plans continuing to provide care to Medicare beneficiaries have higher average performance on HEDIS clinical and ambulatory care access measures than plans that withdrew. PMID- 11880795 TI - Transitions through postacute and long-term care settings: patterns of use and outcomes for a national cohort of elders. AB - OBJECTIVES: Despite the large number of elders using postacute and long-term care services, there is little information about transitions through different settings or the impact of transitions on elders' health. This gap in knowledge is addressed by analyzing the use of postacute and long-term care settings during a 2-year interval by a nationally representative cohort of elders. METHODS: A 2 year longitudinal record of the use of short-stay hospitals and postacute and long-term care settings was constructed for all respondents to the 1994 National Long Term Care Survey age 65 or older in 1992. Indicators of potential transition problems include emergency room visits, potentially avoidable hospital stays, and return to an institutional setting following discharge to the community. RESULTS: Almost 18% of elders, 4.9 million persons, were admitted to or discharged from a study setting between 1992 and 1994. A sizable number of these elders (22.4%) had subsequent health care use, suggesting a possible transition problem. Transitions from acute care hospitals to paid home care represent 20.8% of all transitions and are followed by relatively high rates of potential problems. CONCLUSIONS: This study provides new information on patterns of postacute and long-term care use and the types of transitions most likely to be followed by potential problems. The results suggest three broad strategies for improving the outcome of transitions through postacute and long-term care settings. PMID- 11880796 TI - Risk-adjusted mortality rates as a potential outcome indicator for outpatient quality assessments. AB - OBJECTIVE: The quality of outpatient medical care is increasingly recognized as having an important impact on mortality. We examined whether a clinically credible risk adjustment methodology can be developed for outpatient quality assessments. RESEARCH DESIGN: This study used data from the 1998 National Survey of Ambulatory Care Patients, a prospective monitoring system of outcomes of patients receiving ambulatory care in the Veterans Affairs (VA) integrated service networks. SUBJECTS: Thirty-one thousand eight hundred twenty-three patients were followed for 18 months. MEASURES: The main study outcome measures were observed and risk-adjusted mortality rates. RESULTS: Of the 31,823 patients, 1559 (5%) died during the 18-months of follow-up. Observed mortality rates across the 22 VA integrated service networks varied significantly from 3.3% to 6.7% (P <0.001). Age, gender, comorbidities (Charlson Index), physical health, and mental health were significant predictors of dying. The resulting risk-adjusted mortality model performed well in cross-validated tests of discrimination (c statistic = 0.768; 95% CI, 0.749-0.788) and calibration. Analysis of variance confirmed that the 22 integrated service networks differed in their average level of expected risk (P <0.001). Risk-adjusted rates and ranks of the networks differed considerably from unadjusted ratings. CONCLUSIONS: Risk-adjusted mortality rates may be a useful outcome measure for assessing quality of outpatient care. We have developed a clinically credible risk adjustment model with good performance properties using sociodemographics, diagnoses, and functional status data. The resulting risk adjustment model altered assessments of the performance of the integrated service networks when compared with the unadjusted mortality rates. PMID- 11880797 TI - Continuity of care and clinical effectiveness: outcomes following residential treatment for severe substance abuse. AB - BACKGROUND: Continuity of care (COC) has often been viewed as a crucial indicator of treatment quality for patients with severe psychiatric or addictive disorders. However, the relationship between COC and clinical outcomes has received little empirical evaluation. RESEARCH DESIGN: This study used hierarchical linear modeling to examine the relationship between six indicators of COC and seven outcome measures addressing symptoms, substance abuse, and social functioning. SUBJECTS: Patient interviews were conducted with 1576 veterans 3 months after their discharge from one of 22 residential work therapy programs for the treatment of severe substance abuse. RESULTS: Few significant relationships were found between COC and outcome measures in analyses conducted at both the client and program level and fewer than half of these show better outcomes with greater COC. When a Bonferroni corrected P level of P <0.0012 was used, none of the relationships were statistically significant. CONCLUSION: Although there were significant relationships between outcomes and measures of services received during residential treatment, postdischarge COC does not seem to be related to improved outcomes, at least when examined following long term intensive residential treatment. Thus, our results are specific to the context of aftercare following long-term residential rehabilitation and indicate that the value of standard performance measures may vary by treatment context. PMID- 11880798 TI - Measure-dependent variation in burden of disease estimates: implications for policy. AB - BACKGROUND: Health adjusted life years (HALYs) are used for estimating burden of disease and as outcomes in cost-effectiveness analyses of medical care and public health interventions. OBJECTIVES: The impact of use of health-related quality of life (HRQL) scores associated with the disability-adjusted life year (DALY), the quality-adjusted life year (QALY), and the years of healthy life (YHL) measure on burden of disease estimates by income and race for five illnesses was studied. RESEARCH DESIGN: Abridged life tables were constructed using 1997 death certificate data from the National Center for Health Statistics. These tables were then quality-adjusted using prevalence data from the National Health Interview Survey and HRQL scores obtained using the Quality of Well-Being scale, the DALY, and the YHL measure to estimate burden of disease for five common diseases. Separate estimates were made for low and higher income families as well as black persons and white persons. RESULTS: Measure-related burden of disease estimates differed substantially from one another. Rank order of disease burden was not maintained across measures. Discrepancies in the rank order of disease were greater when different sociodemographic groups were examined. CONCLUSIONS: Diseases and demographic groups will receive differing priorities for intervention or research depending on which measurement system is used to inform decision-making. Refinement and standardization of measures is necessary to enhance their utility for medical care and public health policy applications. PMID- 11880799 TI - Excessive length of iliac arteries in athletes with flow limitations measured by magnetic resonance angiography. AB - PURPOSE: Kinking of the iliac arteries can cause flow limitations in endurance athletes. Such kinking may be treated by surgical release of the iliac arteries. However, when the length of the iliac artery is excessive, this may not be effective. Because threshold values of excessive length of the iliac arteries are unknown, normal values for endurance athletes were obtained and abnormalities encountered in these patients are reported. METHODS: Forty-three endurance athletes (46 symptomatic legs) with flow limitations in the iliac arteries were examined using magnetic resonance angiography (MRA) with hips extended and flexed. The ratio of vessel length to straight-line distance was determined for the common and external iliac arteries. Sixteen national-level cyclists (32 reference legs) served as a control group. RESULTS: For the common iliac artery, length ratios were significantly (P < 0.05) higher in the symptomatic legs than in the reference legs (symptomatic legs: 1.1 +/- 0.12, 1.22 +/- 0.19, reference legs 1.05 +/- 0.04, 1.11 +/- 0.05 with extended and flexed hips, respectively). For the external iliac artery, only in the position with hips flexed, the ratios in the symptomatic legs were significantly higher than in the reference legs (symptomatic legs: 1.11 +/- 0.09, 1.44 +/- 0.23, reference legs 1.08 +/- 0.05, 1.32 +/- 0.13 with extended and flexed hips, respectively). A small proportion of symptomatic legs had extremely high length ratios. CONCLUSION: MRA is effective for determining vessel length. The ratio of vessel length to straight-line distance with extended and flexed hips is a good measure for excessive vessel length and achieves extreme values in a small subgroup of patients. Further prospective study is warranted to define maximal vessel length ratios, which still allow benefit from surgical release of the iliac arteries. PMID- 11880800 TI - Microprocessor-based ambulatory activity monitoring in stroke patients. AB - PURPOSE: Recovery of ambulatory function after stroke is routinely assessed using standardized subject- or observer-rated instruments that do not directly measure ambulatory activities in the home-community setting. Accuracy of conventional pedometers in stroke patients is not established, limiting their application in mobility outcomes monitoring. This study investigates the accuracy and reliability of a mechanical pedometer versus microprocessor-based step activity monitoring (SAM) in gait-impaired hemiparetic stroke patients. METHODS: Accuracy and test-retest reliability of ankle-worn SAM and belt-worn pedometer were tested directly against hand tallied stride counts and cadence during a battery of timed walks in 16 chronic hemiparetic stroke patients. Patients performed replicate 1 min floor walks at self-selected and fastest comfortable paces, and two 6-min walks on separate days. RESULTS: SAM cadence and total stride counts are more accurate than pedometers during 1-min walks at self-selected (99 +/- 1 vs 87 +/- 11.3%, mean +/- SD, P < 0.01); fast pace (98 +/- 2.3% vs 85 +/- 15%, P < 0.01); and repeated 6-min walks performed on separate days (99 +/- 1% vs 89 +/- 12%, P < 0.01). Although SAM is highly reliable (r = 0.97, P < 0.0001) and accurate in all patients under every walking condition tested, the mechanical pedometer demonstrates this high level of accuracy in only half of stroke patients and has poor test-retest reliability (r = 0.64, P < 0.05). CONCLUSION: SAM, but not the conventional pedometer, provides accurate and reliable measures of cadence and total stride counts in hemiparetic stroke patients. Portable microprocessor-based gait monitoring offers potential to quantitatively measure home-community-based ambulatory activity levels in this population. PMID- 11880801 TI - Dental injuries in ice hockey games and training. AB - PURPOSE: The purpose of this study was to determine the incidence, cause, and nature of maxillofacial and dental injuries in ice hockey games and training and also evaluate the use and the effect of dental or facial guards. METHODS: Maxillofacial and dental ice hockey injuries in Finland were studied during years 1991 and 1992. Material was gathered from the insurance company that had practically all the ice hockey licenses in those years. The material consisted of 479 injured ice hockey players who suffered from 650 separate injuries. The most common dental injury was a noncomplicated crown fracture, which accounted for 43.5% of all maxillofacial or dental injuries. Of these noncomplicated crown fractures, almost 70% occurred in the games. RESULTS: The most common cause of accidents was a blow from the ice hockey stick. The stick as a cause of injury was approximately 3 times as common in the games than in training. Only 10% of injured players wore some kind of protective guard. CONCLUSION: A mandatory use of mouthguards and face masks or tightened rules for protection to decrease the high number of maxillofacial and dental injuries in the ice hockey games should be considered. PMID- 11880802 TI - Failure and fatigue characteristics of adhesive athletic tape. AB - PURPOSE: Athletic tape has been commonly reported to lose much of its structural support after 20 min of exercise. Although many studies have addressed the functional performance characteristics of athletic tape, its mechanical properties are poorly understood. This study examines the failure and fatigue properties of several commonly used athletic tapes. METHODS: A Web-based survey of professional sports trainers was used to select the following three tapes for the study: Zonas (Johnson & Johnson), Leukotape (Beiersdorf), and Jaylastic (Jaybird & Mais). Using a hydraulic material testing system (MTS), eight samples of each tape were compared in three different mechanical tests: load-to-failure, fatigue testing under load control, and fatigue testing under displacement control. Differences in tape microstructure were used to interpret the results of the mechanical tests. RESULTS: Significant differences (P < 0.001) in failure load, elongation at failure, and stiffness were found from failure tests. Significant differences were also found (P < 0.001) in fatigue behavior under both modes of control. As a representative example, in one normalized displacement control fatigue test after 20 min of cycling, 21% (Zonas), 29% (Leukotape), and 57% (Jaylastic) of the mechanical support was lost. After cycling, all tapes loaded to failure showed increased stiffness (P < 0.001), indicating significant energy absorption during cycling. Observed differences in the tapes' microstructure were qualitatively consistent with the measured differences in their mechanical properties. CONCLUSION: In understanding the shortcomings of currently available tapes, the results of these tests can now be used as benchmarks with which to compare and develop future tape designs. Ultimately, these improved tapes should reduce ankle injuries among athletes. PMID- 11880803 TI - Epstein-Barr virus reactivation and upper-respiratory illness in elite swimmers. AB - PURPOSE: The aim of this study was to investigate the relationships between latent viral shedding of Epstein-Barr virus (EBV) in saliva, upper-respiratory illness, and mucosal immune suppression in a cohort of highly trained swimmers undertaking intensive training. METHODS: Saliva was collected before selected training sessions from 14 elite male swimmers during a 30-d period of intensive training. Prior infection with EBV was determined by EBV antibody serology. Salivary IgA concentrations were measured by enzyme linked immunosorbent assay (ELISA), and EBV viral shedding (EBV-DNA) was detected by polymerase chain reaction (PCR). Symptoms of upper-respiratory illness were recorded daily. RESULTS: Eleven swimmers (79%) were seropositive for prior EBV infection. Seven EBV seropositive swimmers (64%) had EBV-DNA detected during the study period. Upper-respiratory symptoms (URS) were reported in six of seven swimmers in whom EBV-DNA was detected and in three of four swimmers with no EBV-DNA detection. No URS were reported in the EBV seronegative swimmers. There was a statistically significant relationship between EBV serology status and URS (P = 0.027). EBV-DNA was detected in saliva before the appearance of URS. Salivary IgA levels were significantly lower immediately before the URS (P = 0.01) compared with subsequent peak IgA levels and declined to pre-URS levels on average 11 d after the first appearance of URS. CONCLUSIONS: The time course of appearance of EBV DNA in relation to URS suggests latent viral EBV shedding may be a contributing factor in the URS. The low levels of salivary IgA detected before the URS indicated transient mucosal immune suppression in the study cohort. The viral shedding may alternatively be a reflection of the altered immune control mechanisms that occur in response to intensive exercise and unrelated to the URS. PMID- 11880804 TI - The relationship of heart rate reserve to VO2 reserve in patients with heart disease. AB - Recent reports indicate that among healthy adults, % heart rate reserve (HRR) is more closely related to %VO2 reserve (VO2R) than it is to %VO2max. This finding, in addition to the disparity between %HRR and %VO2max which is greater at low intensities and among low fit individuals, led the American College of Sports Medicine to adopt the use of %VO2R in place of %VO2max when prescribing exercise intensity among healthy adults and persons with heart disease. However, the relationship of %HRR to %VO2R among persons with heart disease has not been described. PURPOSE: Among patients with a myocardial infarction (MI) and patients with chronic heart failure (HF), test the hypothesis that %VO2R is equivalent to %HRR, while %VO2peak is not. METHODS: Using a clinical cardiology outcomes database, symptom-limited treadmill tests from 65 patients with MI and 72 patients with HF were identified. Heart rate and VO2 data were measured continuously and averaged every 15 s. For each subject, linear regression was used to calculate the slope and y-intercept of %HRR versus %VO2R (assuming rest VO2 = 3.5 mL x kg(-1) x min(-1)) and %HRR versus %VO2peak. Mean slope and y intercept were calculated for each group and compared with the line of identity (slope = 1, y-intercept = 0). RESULTS: For the MI and HF groups, the slope of %HRR versus %VO2R was 0.96 +/- 0.02 (+/-SE; P = NS, slope vs 1) and 0.97 +/- 0.02 (P = NS), respectively. And the y-intercept was -1.9 +/- 2.1% (P = NS, y intercept vs 0) and -5.9 +/- 2.1% (P < 0.05) for MI and HF, respectively. For both patient groups, the regression of %HRR versus %VO2peak resulted in a line that differed (P < 0.001) in both slope and y-intercept from the line of identity. CONCLUSIONS: In patients with heart disease, %HRR is a better estimate of %VO2R than %VO2peak. This finding does not affect the current recommended use of %HRR. However, when prescribing exercise based on VO2, relative intensity should be expressed as %VO2R. PMID- 11880805 TI - Fuel oxidation during exercise in middle-aged men: role of training and glucose disposal. AB - PURPOSE: The purpose of this study was to test the hypothesis that carbohydrate (CHO) utilization in middle-aged trained men is increased during hard-intensity exercise and decreased during moderate-intensity exercise in comparison with age matched sedentary men. We also investigated whether a relationship between CHO utilization and glucose disposal exists. METHODS: Seven trained cyclists (Tr) and seven age-matched sedentary men (Sed) underwent an intravenous glucose tolerance test after an overnight fast (minimal model method) to determine their glucose disposal; they also performed two 1-h trials on a cycle ergometer below and above their individual ventilatory threshold (VT). Substrate oxidation was evaluated by indirect calorimetry. Hormonal responses were investigated during exercise. RESULTS: Insulin sensitivity (SI) and glucose effectiveness (Sg) were significantly higher in the Tr group than in the Sed group (P < 0.001, P < 0.03). CHO oxidation was significantly higher in the Tr group than in the Sed group when exercise was performed above VT, whereas CHO oxidation was higher in the Sed group when exercise was performed below VT (P < 0.05). Epinephrine (Epi) response during hard-intensity exercise was higher in the Tr group than in the Sed group (P < 0.01). SI was negatively correlated to CHO oxidation in the Tr group (r = 0.743, P < 0.05). CONCLUSION: Endurance training results in increased CHO utilization during hard-intensity exercise and reduced CHO oxidation during moderate-intensity exercise in middle-aged men. During hard-intensity exercise, the increased CHO utilization in middle-aged trained men is associated with a greater response in Epi and is inversely related with SI. PMID- 11880806 TI - Substrate oxidation is altered in women during exercise upon acute altitude exposure. AB - PURPOSE: The purpose of this study was to determine whether substrate oxidation during submaximal exercise in women is affected by an acute exposure to 4300-m altitude and menstrual cycle phase. METHODS: Eight female lowlanders (mean +/- SD; 33 +/- 3 yr, 58 +/- 6 kg, 163 +/- 8 cm) completed a peak oxygen uptake (VO2peak) and submaximal exercise to exhaustion (EXH) test at 70% of their altitude-specific VO2peak at sea level (SL) and during an acute altitude (AA) exposure to 4300 m in a hypobaric chamber (446 mm Hg) in their early-follicular and midluteal menstrual cycle phase. The respiratory exchange ratio (RER) was calculated from oxygen uptake and carbon dioxide output measurements made during the EXH tests, and used to estimate the percent contribution of fat and carbohydrate to energy metabolism. Blood samples were taken at rest and every 15 min during the EXH tests. Blood samples were evaluated for glucose, lactate, glycerol, free fatty acids, insulin, growth hormone, cortisol, glucagon, epinephrine, norepinephrine, estradiol, and progesterone concentrations. RESULTS: Despite increased (P < 0.05) estradiol and progesterone levels in the midluteal phase, substrate oxidation, energy substrates, and metabolic hormones were not affected by cycle phase at SL or AA. However, free fatty acids and cortisol were increased (P < 0.05) whereas RER was decreased (P < 0.05) during exercise upon AA exposure compared with SL in both cycle phases. CONCLUSIONS: These data suggest that substrate oxidation is altered in women during exercise at AA compared with SL but is not affected by cycle phase. Whether increased fat or protein oxidation accounts for the lower RER values during the AA exposure cannot be determined from this study but warrants further investigation. PMID- 11880807 TI - Leukocyte mitochondria alterations after aerobic exercise in trained human subjects. AB - Exercise is associated with intensity-dependent immune disturbances. Leukocyte mitochondrial alterations and apoptosis may contribute to this phenomenon. PURPOSE: To investigate the effects of different intensities of aerobic exercise (AE) on leukocyte mitochondrial transmembrane potential (MTP) and the propensity of apoptosis. METHODS: Blood samples were collected from 12 subjects who performed AE for 3 consecutive days (35% maximal oxygen consumption (VO2max)). Leukocyte MTP and apoptosis were measured by flow cytometry. The subjects performed two additional sessions of AE of higher intensities (60% and 85% VO2max) with an intervening 4-wk washout period. The measurements were repeated during each session. RESULTS: Leukocyte MTP declined during daily, repetitive AE at an intensity of 60% and 85% VO2max. Similar changes were not found during a more moderate AE (35% VO2max). Leukocytes increased their propensity of apoptosis a period (3-5 d) after the start of the AE. CONCLUSION: High-intensity AE has accumulative effects on the mitochondrial energization status and vitality of peripheral blood leukocytes. Leukocyte MTP is a potentially applicable indicator for monitoring immune distress due to overtraining. PMID- 11880808 TI - Eccentric exercise effect on blood oxidative-stress markers and delayed onset of muscle soreness. AB - PURPOSE: This study examined the effects of a single bout of high-intensity eccentric exercise (EE) on blood protein carbonyls, glutathione status, and muscle damage indicators to ascertain whether blood markers of oxidative stress are elevated at the time delayed onset of muscle soreness (DOMS) occurs. METHODS: Eight healthy men (26.5 +/- 1.5 yr) performed 60 eccentric contractions at approximately 135-150% dominant arm maximum isometric force (MIF) using their nondominant arm elbow flexors. DOMS, range of motion (ROM), MIF, and blood were obtained before, immediately after, and 24, 48, 72, and 96 h after the EE. Blood samples were analyzed for plasma creatine kinase (CK) activity, and protein carbonyls (PC), and erythrocyte glutathione status. RESULTS: A significant decrease in MIF occurred at all times after the EE. ROM decreased from 24 to 96 h, and DOMS increased 24 to 72 h in the nondominant arm as indicated by a repeated measure ANOVA. Plasma CK activity peaked at 72 h (1620 +/- 500 IU x L( 1)) compared with baseline (154 +/- 27 IU x L(-1). Erythrocyte-reduced glutathione (GSH) concentration was not significantly affected by the EE but tended to decrease 23% by 24 h and continued at this level for 96 h. Oxidized glutathione (GSSG) and total glutathione were unchanged over time. A significant increase in plasma PC occurred at 24 and 48 h after eccentric exercise. CONCLUSIONS: The results suggest that 60 EE at 135-150% MIF can result in DOMS, with decreased muscle function and increases in plasma PC at 24 and 48 h without alterations in blood glutathione status. PMID- 11880809 TI - Effect of short-term fat adaptation on high-intensity training. AB - PURPOSE: To determine the effect of short-term (3-d) fat adaptation on high intensity exercise training in seven competitive endurance athletes (maximal O2 uptake 5.0 +/- 0.5 L x min(-1), mean +/-SD). METHODS: Subjects consumed a standardized diet on d-0 then, in a randomized cross-over design, either 3-d of high-CHO (11 g x kg(-1)d(-1) CHO, 1 g x kg(-1) x d(-1) fat; HICHO) or an isoenergetic high-fat (2.6 g CHO x kg(-1) x d(-1), 4.6 g FAT x kg(-1) x d(-1); HIFAT) diet separated by an 18-d wash out. On the 1st (d-1) and 4th (d-4) day of each treatment, subjects completed a standardized laboratory training session consisting of a 20-min warm-up at 65% of VO2peak (232 +/- 23W) immediately followed by 8 x 5 min work bouts at 86 +/- 2% of VO2peak (323 +/- 32 W) with 60-s recovery. RESULTS: Respiratory exchange ratio (mean for bouts 1, 4, and 8) was similar on d-1 for HIFAT and HICHO (0.91 +/- 0.04 vs 0.92 +/- 0.03) and on d-4 after HICHO (0.92 +/- 0.03) but fell to 0.85 +/- 0.03 (P < 0.05) on d-4 after HIFAT. Accordingly, the rate of fat oxidation increased from 31 +/- 13 on d-1 to 61 +/- 25 micromol x kg(-1) x min(-1) on d-4 after HIFAT (P < 0.05). Blood lactate concentration was similar on d-1 and d-4 of HICHO and on d-1 of HIFAT (3.5 +/- 0.9 and 3.2 +/- 1.0 vs 3.7 +/- 1.2 mM) but declined to 2.4 +/- 0.5 mM on d-4 after HIFAT (P < 0.05). Ratings of perception of effort (legs) were similar on d-1 for HIFAT and HICHO (14.8 +/- 1.5 vs 14.1 +/- 1.4) and on d-4 after HICHO (13.8 +/- 1.8) but increased to 16.0 +/- 1.3 on d-4 after HIFAT (P < 0.05). CONCLUSIONS: 1) competitive endurance athletes can perform intense interval training during 3-d exposure to a high-fat diet, 2) such exercise elicited high rates of fat oxidation, but 3) compared with a high-carbohydrate diet, training sessions were associated with increased ratings of perceived exertion. PMID- 11880811 TI - Normative database of isometric cervical strength in a healthy population. AB - OBJECTIVES: This was a descriptive study involving maximal and average isometric strength measurements of the cervical musculature. The purpose of this study was 1) to determine the test-retest reliability of a computerized dynamometer for measuring muscular strength of the neck flexor and extensor muscles in a healthy population, and 2) to afford a normative database of the maximal and averaged isometric strength of the cervical flexors and extensors in a healthy population of 94 volunteers. The literature contains only a few descriptive studies pertaining to strength levels of the cervical musculature; none of these studies used a computerized dynamometer to determine cervical isometric strength. Only a few studies include cervical flexor and extensor isometric strength, evaluating its normal ratio as well as their correlation to body mass index. METHODS: Testing was carried out using a computerized dynamometer to measure isometric cervical strength at 0 degrees (neutral), 5 degrees, and 10 degrees of flexion and extension of the neck. RESULTS: Men show approximately 30 to 40% more strength than women both for flexion and extension at all angles and age groups. Neutral positioning showed the maximum strength values for both flexion and extension. CONCLUSION: There was a significant negative correlation between age and cervical strength and a significant positive correlation between weight and strength and between height and strength. However, the flexor/extensor ratio was maintained around 0.6 in all ages. PMID- 11880810 TI - Central and peripheral cardiovascular adaptations during a maximal cycle exercise in boys and men. AB - PURPOSE: Stroke volume response to exercise depends on changes in cardiac filling, intrinsic myocardial contractility, and left ventricular afterload. The purpose of this study was to compare these responses during an upright cycle test performed until exhaustion in children and adults. METHODS: Stroke volume, cardiac output (Doppler echocardiography), left ventricular dimensions (two dimensional and time-movement echocardiography), as well as arterial pressure and systemic vascular resistance (SVR) were assessed in 17 boys (mean age, 11.7 +/- 0.6 yr) and 23 young adult men (mean age, 21.2 +/- 2.7 yr) having a similar aerobic potential. All variables were measured at the end of the resting period, during the final minute of each workload, and during the last minute of the test. RESULTS: No significant differences were obtained for stroke volume, cardiac output, and left ventricular dimensions when they were scaled to body surface area at rest and whatever the exercise intensity. However, arteriovenous oxygen uptake was higher and the SVR lower in the adults than in the children. CONCLUSION: The patterns of stroke volume, as well as its underlying mechanisms, were not age-related during an upright maximal exercise test. However, other studies are required to understand further the effect of pubertal status on the peripheral cardiovascular system. PMID- 11880812 TI - Rate and amplitude of adaptation to intermittent and continuous exercise in older men. AB - PURPOSE: This study determined the amplitude and rate of adaptation to 10 wk of continuous (CEx) and intermittent exercise (IEx) in a group of older men when the training intensity and total amount of work completed by each exercise group were the same. METHODS: Ten healthy men were assigned to either a CEx (63 +/- 1 yr) or IEx (65 +/- 1 yr) group while a further five subjects (65 +/- 1 yr) acted as nonexercising controls (CON). The three groups (CEx, IEx, and CON) were matched for age, peak oxygen uptake (VO2peak), and cardiac output (Qpeak) before commencing training. The CEx group trained for 30 min at an intensity corresponding to 70-75% VO2peak, and the IEx group trained for a total exercise time of 30 min using intermittent exercise (60-s exercise, 60-s rest) at the same absolute intensity as the CEx group (CEx 112 +/- 5W; IEx 112 +/- 5W). The exercise groups trained three times per week and completed a similar amount of work during each training session (CEx, 199 +/- 9 kJ; IEx 195 +/- 9 kJ, P = 0.67). RESULTS: The CEx and IEx groups had similar and significant amplitude increases in peak VO2, ventilation (VEpeak), power, Q, and SV after training. Peak VO2, Qpeak, SVpeak, and peak arteriovenous O2 difference for the CON group were unchanged. The change in VO2peak, peak ventilation, and peak power for CEx and IEx groups were best described by a linear model. Moreover, the CEx and IEx groups had the same rate of change in VO2peak (CEx: 0.02 +/- 0.00 L x min(-1) x wk(-1), IEx: 0.02 +/- 0.00 L x min(-1) x wk(-1), P = 0.32), VEpeak (CEx: 2.0 +/- 0.2 L x min(-1) x wk(-1), IEx: 1.2 +/- 0.5 L x min(-1) x wk(-1), P = 0.10), and peak power (CEx: 2.6 +/- 0.4 W x wk(-1), IEx: 2.6 +/- 0.4 W x wk(-1), P = 0.92). CONCLUSION: These results suggest that the amplitude and rate of change of select adaptations in men aged 60-70 yr are independent of the mode of training (i.e., continuous or intermittent exercise) when the absolute training intensity and the total amount of work completed were similar. PMID- 11880813 TI - Effects of power and flexibility training on vertical jump technique. AB - PURPOSE: The purpose of this study was to assess the effects of power and flexibility training on countermovement and drop jump techniques. METHOD: All jumps were executed with the goal of attaining maximum height and no restrictions were placed on the magnitude of countermovement or ground contact time. Subjects underwent initial testing followed by random allocation to one of four groups: power training to increase vertical jump height (P), stretching to increase flexibility (S), a combination of power and stretch training (PS), and a control group (C). Training lasted for 10 wk, followed by retesting. Jump height was calculated in addition to the following technique variables: eccentric lower-limb stiffness produced during the countermovement phase, magnitude of countermovement, and in the case of the drop jumps, ground contact time. RESULTS: Groups PS, P, and S all increased countermovement jump (CMJ) height, but only groups PS and P increased drop jump height (DJ30, DJ60, and DJ90 for drop jumps performed from 30-, 60-, and 90-cm drop heights). The technique changes associated with power training were increases in magnitude of countermovement (CMJ, DJ30, DJ60, and DJ90) and increases in ground contact time (DJ30 and DJ60). In addition, the eccentric lower-limb stiffness produced during the countermovement phase of the jumps increased for CMJ and decreased for DJ30, DJ60, and DJ90. Stretching appeared to have no significant effect on CMJ or drop jump technique. CONCLUSION: The results of this study show that when the training goal is maximum jump height alone, it is likely that drop jump technique will change in the direction of a lower eccentric leg stiffness, greater depth of countermovement, and a longer ground contact time, whereas for a countermovement jump eccentric leg stiffness and the depth of countermovement will both increase. It is proposed that these technique changes are a result of attempting to optimize a complex combination of factors involved in jumping (e.g., utilization of elastic energy, Golgi tendon organ inhibition, and contractile component contribution). PMID- 11880814 TI - Factors that alter body fat, body mass, and fat-free mass in pediatric obesity. AB - PURPOSE: The purpose of this study was to quantify the effects of exercise treatment programs on changes in body mass, fat-free mass, and body fat in obese children and adolescents. METHODS: By using the meta-analytic approach, studies that met the following criteria were included in our analyses: 1) at least six subjects per group; 2) subject groups consisting of children in the 5- to 17-yr age range; 3) pretest and posttest values for either body mass, percent body fat, or fat-free mass (FFM); 4) used exercise as a mode of treatment (e.g., walking, jogging, cycle ergometry, high-repetition resistance exercise, and combinations); 6) training programs >or= 3 wk; 7) full-length publications (not conference proceedings); 8) apparently "healthy" children (i.e., free from endocrine diseases and disorders); and 9) published studies in English language journals only. RESULTS: A total of 120 investigations were located that addressed the issue of exercise as a method of treatment in pediatric obesity. Of those, 30 met our criteria for inclusion. Across all designs and categories, fixed-effects modeling yielded significant decreases in the following dependent variables: 1) percent body fat (mean = 0.70 +/- 0.35; 95% CI = 0.21 to 1.1); 2) FFM (mean = 0.50 +/- 0.38; 95% CI = 0.03 to 0.57); 3) body mass (mean = 0.34 +/- 0.18; 95% CI = 0.01 to 0.46); 4) BMI (mean = 0.76 +/- 0.55; 95% CI = 4.24 to 1.7), and 5) VO2max (mean = 0.52 +/- 0.16; 95% CI = 0.18 to 0.89), respectively. Significant differences were found as a function of the type intervention groups (exercise vs exercise + behavioral modification; P < 0.04); body composition assessment methods (skinfold vs hydrostatic weighing, DEXA, and total body water; P < 0.006); exercise intensity (60-65%, vs >or= 71% VO2max; P < 0.01); duration ( 30 min; P < 0.03); and mode (aerobic vs aerobic + resistance training; P < 0.02). Stepwise linear regression suggested that initial body fat levels (or body mass), type of treatment intervention, exercise intensity, and exercise mode accounted for most of the variance associated with changes in body composition after training. CONCLUSIONS: Exercise is efficacious for reducing selected body composition variables in children and adolescents. The most favorable alterations in body composition occurred with 1) low-intensity, long duration exercise; 2) aerobic exercise combined with high-repetition resistance training; and 3) exercise programs combined with a behavioral-modification component. PMID- 11880815 TI - Comparison of methods for assessing body composition changes during weight loss. AB - PURPOSE: Four cross-sectional studies have reported that percent body fat (%BF) measured by dual-energy x-ray absorptiometry (DXA) is significantly higher compared with values obtained with air displacement plethysmography (ADP) using the Bod Pod(R) in normal-weight individuals. This study was performed to confirm these findings in an overweight population and to assess whether DXA and ADP detected similar changes in body composition after moderate weight loss. METHODS: Twelve women (42 +/- 8 yr) and 10 men (40 +/- 11 yr) had their %BF, fat mass (FM), and fat-free mass (FFM) measured using DXA and ADP before and after an 8-wk weight-loss program involving moderate energy restriction and exercise. RESULTS: Body weight decreased significantly in women (-4.3 +/- 3.4 kg) and men (-4.7 +/- 3.1 kg). There were significant method (ADP vs DXA) and time (pre and post) effects but no method by time or gender interactions. Methods were significantly different in estimating %BF, FM, and FFM with ADP estimates of %BF and FM being lower and estimates of FFM higher than corresponding DXA values (P = 0.000). There were significant correlations accounting for a high degree of the shared variance between DXA and ADP (r = 0.98 to 0.99) for %BF, FM, and FFM and lower correlations for the changes in %BF (r = 0.66), FM (r = 0.86), and FFM (r = 0.34). In response to weight loss, the mean changes in %BF, FM, and FFM were not significantly different between methods (P > 0.05). CONCLUSION: Both DXA and ADP measure changes in body composition after small to moderate weight loss to the same extent and with similar sensitivity. PMID- 11880816 TI - Intraindividual allometric development of aerobic power in 8- to 16-year-old boys. AB - PURPOSE: The aims of this study are two-fold: first, to analyze intraindividual allometric development of aerobic power of 73 boys followed at annual intervals from 8 to 16 yr, and second, to relate scaled aerobic power with level of habitual physical activity and biological maturity status. METHODS: Peak VO2 (treadmill), height, and body mass were measured. Biological maturity was based on age at peak height velocity (PHV) and level of physical activity was based on five assessments between 11 and 15 yr and at 17 yr. Interindividual and intraindividual allometric coefficients were calculated. Multilevel modeling was applied to verify if maturity status and activity explain a significant proportion of peak VO2 after controlling for other explanatory characteristics. RESULTS: At most age levels, interindividual allometry coefficients for body mass exceed k = 0.750. Intraindividual coefficients of peak VO2 by body mass vary widely and range from k' = 0.555 to k' = 1.178. Late maturing boys have smaller k' coefficients than early maturing boys. CONCLUSION: Peak VO2 is largely explained by body mass, but activity level and its interaction with maturity status contribute independently to peak VO2 even after adjusting for body mass. PMID- 11880817 TI - Neuromuscular adaptations to concurrent strength and endurance training. AB - PURPOSE: The purpose of this study was to examine muscle morphological and neural activation adaptations resulting from the interaction between concurrent strength and endurance training. METHODS: Thirty sedentary healthy male subjects were randomly assigned to one of three training groups that performed 10 wk of 3-d x wk(-1) high-intensity strength training (S), cycle endurance training (E), or concurrent strength and endurance training (CC). Strength, quadriceps-muscle biopsies, computed tomography scans at mid-thigh, and surface electromyogram (EMG) assessments were made before and after training. RESULTS: S and CC groups demonstrated similar increases (P < 0.0001) in both thigh extensor (12 and 14%) and flexor/adductor (7 and 6%) muscle areas. Type II myofiber areas similarly increased (P < 0.002) in both S (24%) and CC (28%) groups, whereas the increase (P < 0.004) in Type I area with S training (19%) was also similar to the nonsignificant (P = 0.041) increase with CC training (13%). Significant increases (P < 0.005) in maximal isometric knee-extension torque were accompanied by nonsignificant (P 0.38) in the EMG/torque relation across 20 to 100% maximal voluntary contractions occurred in any group. A small 3% increase (P < 0.01) in thigh extensor area was the only change in any of the above variables with E training. CONCLUSIONS: Findings indicate 3-d x wk(-1) concurrent performance of both strength and endurance training does not impair adaptations in strength, muscle hypertrophy, and neural activation induced by strength training alone. Results provide a physiological basis to support several performance studies that consistently indicate 3-d x wk( 1) concurrent training does not impair strength development over the short term. PMID- 11880818 TI - Changes in blood volume and oxygenation level in a working muscle during a crank cycle. AB - PURPOSE: This study examined circulatory and metabolic changes in a working muscle during a crank cycle in a pedaling exercise with near-infrared spectroscopy (NIRS). METHODS: NIRS measurements sampled under stable metabolic and cadence conditions during incremental pedaling exercise were reordered according to the crank angles whose signals were obtained in eight male subjects. RESULTS: The reordered changes in muscle blood volume during a crank cycle demonstrated a pattern change that corresponded to changes in pedal force and electrical muscle activity for pedal thrust. The top and bottom peaks for muscle blood volume change at work intensities of 180 W and 220 W always preceded (88 +/ 32 and 92 +/- 23 ms, respectively) those for muscle oxygenation changes. Significant differences in the level of NIRS parameters (muscle blood volume and oxygenation level) among work intensities were noted with a common shape in curve changes related to pedal force. In addition, a temporary increase in muscle blood volume following a pedal thrust was detected at work intensities higher than moderate. This temporary increase in muscle blood volume might reflect muscle blood flow restriction caused by pedal thrusts. CONCLUSION: The results suggest that circulatory and metabolic conditions of a working muscle can be easily affected during pedaling exercise by work intensity. The present method, reordering of NIRS parameters against crank angle, serves as a useful measure in providing additional findings of circulatory dynamics and metabolic changes in a working muscle during pedaling exercise. PMID- 11880820 TI - Influence of cycling cadence on subsequent running performance in triathletes. AB - PURPOSE: The purpose of this study was to investigate the influence of different cycling cadences on metabolic and kinematic parameters during subsequent running. METHODS: Eight triathletes performed two incremental tests (running and cycling) to determine maximal oxygen uptake (VO2max) and ventilatory threshold (VT) values, a cycling test to assess the energetically optimal cadence (EOC), three cycle-run succession sessions (C-R, 30-min cycle + 15-min run), and one 45-min isolated run (IR). EOC, C-R, and IR sessions were realized at an intensity corresponding to VT + 5%. During the cycling bouts of C-R sessions, subjects had to maintain one of the three pedaling cadences corresponding to the EOC (72.5 +/- 4.6 rpm), the freely chosen cadence (FCC; 81.2 +/- 7.2 rpm), and the theoretical mechanical optimal cadence (MOC, 90 rpm; Neptune and Hull, 1999). RESULTS: Oxygen uptake (VO2) increased during the 30-min cycling only at MOC (+12.0%) and FCC (+10.4%). During the running periods of C-R sessions, VO2, minute ventilation, and stride-rate values were significantly higher than during the IR session (respectively, +11.7%, +15.7%, and +7.2%). Furthermore, a significant effect of cycling cadence was found on VO2 variability during the 15-min subsequent run only for MOC (+4.1%) and FCC (+3.6%). CONCLUSION: The highest cycling cadences (MOC, FCC) contribute to an increase in energy cost during cycling and the appearance of a VO2 slow component during subsequent running, whereas cycling at EOC leads to a stability in energy cost of locomotion with exercise duration. Several hypotheses are proposed to explain these results such as changes in fiber recruitment or hemodynamic modifications during prolonged exercise. PMID- 11880821 TI - Creatine supplementation improves muscular performance in older men. AB - PURPOSE: Creatine supplementation has been shown to enhance muscle strength and power after only 5-7 d in young adults. Creatine supplementation could therefore benefit older individuals because aging is associated with a decrease in muscle strength and explosive power. METHODS: We examined the effects of 7 d of creatine supplementation in normally active older men (59-72 yr) by using a double-blind, placebo-controlled design with repeated measures. After a 3-wk familiarization period to minimize learning effects, a battery of tests was completed on three occasions separated by 7 d (T1, T2, and T3). After T1, subjects were matched and randomly assigned into creatine (N = 10) and placebo (N = 8) groups. After T2, subjects consumed supplements (0.3 g x kg(-1) x d(-1)) for 7 d until T3. All subjects were tested for maximal dynamic strength (one-repetition maximum leg press and bench press), maximal isometric strength (knee extension/flexion), upper- and lower-body explosive power (6 x 10-s sprints on a cycle ergometer), and lower-extremity functional ability (timed sit-stand test and tandem gait test). Body composition was assessed via hydrostatic weighing, and blood samples were obtained to assess renal and hepatic responses and muscle creatine concentrations. RESULTS: No significant increases in any performance measures were observed from T1 to T2 with the exception of isometric right-knee flexion in the placebo group indicating stability in the testing protocols. Significant group-by -time interactions indicated the responses from T2 to T3 were significantly greater (P 24 hours had significantly elevated pulse rate, temperature, leukocyte count, and C reactive protein (CRP) and lower TAC. No significant difference was found in age, sex, temperature, respiratory rate, blood pressure, and MDA level. Multivariate logistic regression analysis revealed that CRP and TAC were significant indicators of quantitative variables for disposition. This study found a correlation exists between oxidative stress and disease severity in patients with abdominal pain. This suggests that TAC might be useful as a guide for patient disposition in the emergency department. PMID- 11880868 TI - Parenteral analgesic and sedative use among ED patients in the United States: combined results from the National Hospital Ambulatory Medical Care Survey (NHAMCS) 1992-1997. AB - The objective of the study was to describe parenteral analgesic and sedative (PAS) use among patients treated in US emergency departments (EDs). Data representing 6 consecutive years (1992-1997) from the National Hospital Ambulatory Medical Care Survey (NHAMCS) were combined and analyzed. Patients were identified as having received PAS if they received fentanyl, ketamine, meperidine, methohexital, midazolam, morphine, nitrous oxide, or propofol. Patients were stratified according to age (pediatric <18 years), race, gender, insurance, type of hospital, urgency of visit, and ICD-9 diagnostic codes. Logistic regression was performed to determine independent associations and calculate odds ratios (OR) for receiving analgesia or sedation. A total of 43,725 pediatric and 114,207 adult ED encounters were analyzed and represented a weighted sample of 555.3 million ED visits. For patients with orthopedic fractures, African American children covered by Medicaid insurance were the least likely to receive PAS (OR 0.2, 95% confidence interval 0.1-0.6). These results suggest that variations may be occurring among ED patients receiving PAS. PMID- 11880869 TI - Stability of diazepam rectal gel in ambulance-like environments. AB - The objective of the present study was to evaluate the stability of diazepam rectal gel (Diastat) in various conditions of temperature and light exposure as might be found in ambulances. Three lots of Diastat (Xcel Pharmaceuticals, San Diego, CA) in various fill/syringe configurations were evaluated in controlled conditions of a freeze-thaw cycle, hard freeze (-30 degrees C for 72 hours), extreme light exposure (1,000 ft candles for 1 month), and long-term evaluation at either 30 degrees C or 40 degrees C. In the various configurations and tests, diazepam concentration always exceeded 95% of label, with no changes of note in excipients or physicochemical properties. The estimated shelf-life at 30 degrees C exceeds 48 months. Based on the results of the present study, the restocking frequency of Diastat in ambient storage conditions (eg, ambulances), could be up to 48 months in nonfreezing environments, as long as this does not exceed the labeled expiration date on the product. PMID- 11880870 TI - Active epistaxis at ED presentation is associated with arterial hypertension. AB - Epistaxis and hypertension are frequent in the general population, but an association is still controversial. Aim of this retrospective cohort study was to test if active epistaxis at emergency department (ED) presentation is associated with hypertension. Patients with active epistaxis at ED presentation (n = 271; 73%) were compared with controls without active bleeding (n = 101; 27%). By multivariate logistic regression modeling we found that active epistaxis was independently associated with history of hypertension (odds ratio 2.8 [95% CI 1.4 to 5.6; P =.004] adjusted for age, gender, rhinitis, diffuse bleeding, and malignant diseases). Patients with active epistaxis had higher blood pressure at presentation compared with controls (systolic blood pressure 165 v 153 mmHg, P <.001, diastolic blood pressure 85 v 77 mmHg, P <.001). Active epistaxis at ED presentation is associated with arterial hypertension. PMID- 11880871 TI - Removal of esophageal foreign bodies in the pediatric ED: is ketamine an option? AB - The objective of the study was to evaluate our experience with esophageal foreign body (EFB) removal comparing the use of ketamine-midazolam (K-M) and fentanyl midazolam (F-M) in the emergency department (ED), to admission and general anesthesia (GA) in the operating room (OR). A retrospective review of all children undergoing EFB removal at our institution during a 2-year period was conducted. A total of 93 patients were identified: K-M 57/93 (61.2%), F-M 28/93 (30.1%), GA 5/93 (5.4%), and 3/93 (3.2%) by other means. Mean procedure durations were 4.8 min for K-M and 7.0 min for F-M. Mean lengths of stay (LOS) for ED procedures were 3.6 hrs for K-M and 5.7 hrs for F-M, versus 17.7 hrs if admitted. Transient hypoxemia occurred in 10.7% of K-M and 15.4% of F-M. Removal of EFBs in the ED may obviate the need for admission. In our experience, the use of K-M is associated with fewer airway complications, shorter removal times, and an overall shorter LOS. PMID- 11880872 TI - Validation of a brief screening tool to detect depression in elderly ED patients. AB - Although depression is the most common psychiatric disorder in the elderly, it is often unrecognized by physicians. The objective of the study was to assess the utility of a 3-question screening instrument (ED-DSI) to detect depression among elderly emergency department (ED) patients. We used a prospective convenience sample of English-speaking ED patients >or=65yr. Patients were excluded for being too ill to participate, having obvious dementia, or acute changes in mentation. A physician administered the ED-DSI. Trained research personnel blinded to ED-DSI answers then administered the 30 question Geriatric Depression Scale (GDS). The ED-DSI was considered positive if the patient answered yes to one or more questions and subjects were considered depressed when the GDS score was >or=10. ED-DSI was compared to GDS using contingency tables. A total of 103 subjects were enrolled. Average age was 75 years and subjects were predominately female (66%) and white (85%). GDS identified 33 patients (32%) as depressed. Of these, 26 were correctly identified by ED-DSI giving an ED-DSI sensitivity of 79% (95%CI (65%, 93%)) specificity of 66% (95%CI (54%, 78%)) and a negative predictive value of 87% (95%CI (79%, 95%)). The 3-question ED-DSI is a useful tool to detect depression in this population of ED patients. PMID- 11880873 TI - Nasal positive-pressure technique for nasal foreign body removal in children. AB - Nasal foreign bodies in children are often encountered in emergency medicine and many methods of removal exist. This study examines the nasal positive pressure technique, a method not previously described in the literature. A retrospective chart review of nasal foreign body removal using the nasal positive pressure technique was conducted, as well as, a follow-up telephone survey of patient's parents. Nine patients were identified over a 1-year period. The average time from triage to discharge was 34 minutes (15-106 minutes). No sedation, consultation or complications were recorded. Five of the 9 parents completed the survey. All 5 (100%) described the procedure less traumatic than an injection, and 4 (80%) described the procedure similar or less traumatic than an oral pharyngeal exam with a tongue depressor. Nasal positive pressure technique for nasal foreign body removal is a safe and efficient method, and it is relatively nontraumatic in the view of parents. PMID- 11880874 TI - Intraperitoneal blood missed on a FAST examination using portable ultrasound. AB - Bedside ultrasonography is rapidly gaining popularity in the evaluation of emergency patients. Several manufacturers have developed hand-held ultrasound machines that make the technology easy to transport and available in settings where such diagnostic tests were previously unobtainable. The miniaturization of equipment often means compromises and no studies exist comparing the high quality imaging capabilities of larger conventional ultrasound units to hand-held machines on actual patients. We present 3 cases in which intra-abdominal fluid stripes, important markers of intraperitoneal bleeding, were not visible with a popular hand-held unit, but were identified with a larger mobile ultrasound machine. These findings should caution emergency physicians to be aware of this limitation along with the many advantages of these new and popular hand-held ultrasound units. PMID- 11880875 TI - Increasing importance of the elderly in a trauma system. AB - The objectives of the study were to determine age- and sex-related trends in hospitalization after injuries. Statewide hospitalization data from 1980-1999 were evaluated annually by sex and age (0-39, 40-64, 65-99), incidence, mean length of stay (LOS), and disposition. Males 0-39 represented 39.6% of admissions in 1980, but only 20.7% in 1999; females >or=65 are now most frequent. These trends partly reflect population changes, but rates of hospitalization for young men have also declined. For age < 65, LOS has gradually decreased since 1980; for age >or=65, LOS decreased after 1990. However, most elderly patients were formerly discharged home, whereas now most are discharged to skilled nursing facilities (SNF). The proportion of younger patients hospitalized in trauma centers increased significantly, but over age 65 this proportion remained stable. Major changes have occurred in populations served by trauma systems. Preventing injuries in the elderly is increasingly important. Accurate evaluation of cost effectiveness in the elderly must include outcomes from SNF. PMID- 11880876 TI - Reagent strip diagnosis of UTI in a high-risk population. AB - A study was undertaken to determine the accuracy of reagent strip diagnosis of urinary tract infection (UTI) in patients seen in the ED who were subsequently admitted to hospital with this diagnosis. The reagent strip results of 100 patients were recorded and the sensitivity and specificity of various parameters in the diagnosis of UTI were calculated. Our results indicate that even in a group of patients with high clinical probability of UTI and with symptoms severe enough to be hospitalized, reliance on positivity of any single parameter alone gives unacceptably low sensitivity: 89%, 39%, 68%, and 63% for WBC, nitrite, RBC, and protein, respectively. We conclude that dipstick analysis is not sufficiently sensitive for diagnosis of UTI in high risk patients in whom missed diagnosis would have serious consequences. PMID- 11880877 TI - Orthopedic pitfalls in the ED: fight bite. AB - Clenched fist injuries to the mouth ("fight bite") are notorious for being the worst human bites. These are often treated as minor injuries, without the recognition that the joint capsule, the extensor tendon, or the deep fascial spaces may have been violated and contaminated with oral bacteria. Significant morbidity can result from late presentation or inadequate initial management. The emergency physician needs to remain vigilant for complications associated with closed fist injury. This review article examines the clinical presentation, diagnostic techniques, and management options applicable to the emergency physician in the treatment of fight bite. PMID- 11880878 TI - The feasibility of full computerization in the ED. AB - For understanding the feasibility of full computerization of an emergency department (ED), we investigated the completion rate performed by doctors, nurses, or registration clerks since the implementation of full computerization in our ED. We evaluated the changing style of chart-recording, from hand-writing pattern to full computer recording, by recording the execution rate of different information keyed by doctors, nurses, or registration clerks according to their work in ED. We recorded and analyzed different monthly reports of the execution rate in the 18-month period of study. Statistical analysis was performed using Wilcoxon rank-sum test or Kruskal-Wallis one-way ANOVA. The average monthly census was 4570.1 +/- 580.7 (95% confidence interval [CI] for mean: 4281.3, 4858.9). The average execution rate for mode of arrival and triage classification were 97.1 +/- 4.1% (95% CI for mean: 95.1%, 99.1%) and 97.2 +/- 4.1% (95% CI for mean: 95.2%, 99.2%), respectively. In comparison with the execution rate for disposition status between the period of the first 10 months (keying data by nurses) and the late 8 months (keying data by clerks), it showed 72.0 +/- 33.2% v 96.7 +/- 2.0%; 66.7 +/- 35.0% v 95.8 +/- 1.9%; 57.5 +/- 32.0% v 88.2 +/- 8.2% in nontrauma, trauma and pediatric section, respectively, with statistic significance (P <.01). To compare the rate of execution performed by physicians, we divided the study period into 3 phases (phase 1: first 6 months, phase 2: 7-12 months, phase 3: 13-18 months of the study period). The results were statistically significant (P =.004) in phase 3 (83.4 +/- 5.3%) with higher execution rate than phase 1 (69.7 +/- 7.7%) and phase 2 (75.2 +/- 4.9%) in trauma physician. In the pediatric section, it was also significantly higher in phase 3 than phase 2 (88.2 +/- 7.7% v 70.7 +/- 5.9%, P =.012). We concluded that it is efficient to key in data by registration clerks instead of nurses, and it takes time to persuade and educate most physicians to cooperate in using the computer while seeing patients. PMID- 11880879 TI - Emergency health services in Bulgaria. AB - The emergency care system in Bulgaria is evolving as a hybrid of the former "Soviet-style" health service and western-style emergency medicine. Bulgaria like other "Eastern bloc" Communist nations has undergone a sweeping socioeconomic transformation during the past 10 years. These changes have had profound consequences including the development of emergency services and the recognition of emergency medicine as a specialty in Bulgaria. PMID- 11880880 TI - Risk factors in oligoanalgesia. PMID- 11880881 TI - Simultaneous bilateral posterior dislocation of shoulder. PMID- 11880882 TI - Diagnostic accuracy of a rapid antigen test for GABHS performed by nurses in a pediatric ED. PMID- 11880883 TI - Methemoglobinemia with aluminum phosphide poisoning. PMID- 11880884 TI - Goal-directed sonography in the ED. PMID- 11880885 TI - Hemorrhagic corpus luteum presenting as acute gastroenteritis. PMID- 11880886 TI - Atlanto-occipital traumatic dislocation. PMID- 11880887 TI - Adenosine-induced ventricular flutter in an ethanol-intoxicated patient. PMID- 11880888 TI - Abdominal crisis caused by perforation of ileal lymphoma. PMID- 11880889 TI - Emergency abdominal surgery for sigmoid mesocolic hernia. PMID- 11880890 TI - Increase in ferric and ferrous iron in the rat hippocampus with time after kainate-induced excitotoxic injury. AB - The present study aimed to elucidate the distribution of ferric and ferrous iron in the hippocampus after kainate-induced neuronal injury. A modified Perl's or Turnbull's blue histochemical stain was used to demonstrate Fe3+ and Fe2+ respectively. Very light staining for iron was observed in the hippocampus, in normal or saline-injected rats and 1-day post-kainate-injected rats. At 1 week postinjection, a number of Fe3+-positive, but very few Fe2+-positive, cells were present, in the degenerating CA fields. At 1 month postinjection, large numbers of Fe3+-positive glial cells, and some Fe2+-positive blood vessels, were observed. At 2 months postinjection, large numbers of Fe3+- and Fe2+-positive glial cells were present. The labeled cells had light and electron microscopic features of oligodendrocytes, and were double labeled with CNPase, a marker for oligodendrocytes. The observation of an increasing number of Fe3+- and Fe2+ positive cells in the degenerating hippocampus with time is consistent with the results of a nuclear microscopic study, in which an increasing amount of iron was detected in the degenerating hippocampus after kainate injection. In addition, the present study showed a shift in the oxidation state of the accumulated iron, with more cells becoming Fe2+ at a late stage. A possible consequence of the high amounts of Fe2+ in the hippocampus after kainate injection is that it could promote free radical damage in the lesioned areas. PMID- 11880891 TI - Effects of changes in hip joint angle on H-reflex excitability in humans. AB - We examined the amplitude modulation of the soleus (Sol) H-reflex during controlled variations of the hip joint angle in 21 healthy adult human subjects. Hip angle variations were imposed separately, or in combination either with stimulation of the plantar skin or with electrical activation of muscle afferents from the medial gastrocnemius (MG) nerve. We found that with subjects in the supine position, flexion of the hip significantly depressed Sol H-reflex excitability, by as much as 50% of control reflex values (Ho) recorded at 10 degrees of hip flexion. Conversely, significant facilitation of the H-reflex was observed when the hip joint was extended (10 degrees), with amplitudes reaching 200+/-15.3% of Ho. Changes in H-reflex amplitude were also observed during electrical stimulation of either the foot sole or the MG nerve, when stimuli were delivered at different hip angles. Foot sole stimulation resulted in facilitation of the H-reflex with the hip extended while depression of the reflex was recorded with the hip flexed. In contrast, MG nerve stimulation at group-I muscle afferent strength resulted in a significant increase in the Sol H-reflex magnitude with the hip flexed, while during hip extension, no significant effects were observed [corrected]. This study provides evidence for the existence of a spinal mechanism, determined principally by the hip joint angle, which promotes switching between inhibitory and facilitatory pathways during hip flexion and extension. The origins of such a spinal mechanism are discussed. PMID- 11880892 TI - Fear of falling modifies anticipatory postural control. AB - This study investigated the influence of fear of falling or postural threat on the control of posture and movement during a voluntary rise to toes task for 12 healthy young adults. Postural threat was modified through alterations to the surface height at which individuals stood (low or high platform) and changes in step restriction (away from or at the edge of the platform) creating four levels of postural threat: LOW AWAY, LOW EDGE, HIGH AWAY and HIGH EDGE. To rise to the toes, an initial postural adjustment must destabilise the body so that it can be moved forward and elevated to a new position of support over the toes. Centre of pressure and centre of mass profiles, as well as tibialis anterior (TA), soleus (SO) and gastrocnemius (GA) muscle activity patterns were used to describe this behaviour. The results showed that the performance of the rise to toes task was significantly modified when positioned at the edge of the high platform. In this situation, the central nervous system reduced the magnitude and rate of the postural adjustments and subsequent voluntary movement. Although the duration of the movement was lengthened for this most threatening condition, the sequencing and relative timing of TA, SO and GA muscle activity was preserved. These changes in rise to toes behaviour were accompanied by evidence of increased physiological arousal and participant reports of decreased confidence, increased anxiety and decreased stability. Evidence of fear of falling effects on anticipatory postural control is clinically relevant as it may explain deficits in this control observed in individuals with balance disorders. For example, individuals with Parkinson's disease or cerebellar dysfunction demonstrate impaired performance on the rise to toes task as reflected in alterations of both the timing and magnitude of their anticipatory postural adjustments. Our findings suggest alterations in the magnitude of postural adjustments may be magnified by fear of falling while changes in the timing of postural adjustments may reflect underlying pathology. PMID- 11880893 TI - Use of the trunk for reaching targets placed within and beyond the reach in adult hemiparesis. AB - Multijoint movements such as reaching are impaired after brain lesions involving sensorimotor areas and pathways. However, the mechanisms by which such lesions affect motor control are not fully understood. Direct effects of the lesion may be partly compensated by both the system's redundancy and its plasticity. Indeed stroke patients with limited arm movement can reach objects placed within the reach of the arm by using a compensatory strategy involving trunk recruitment. A similar strategy is observed in healthy individuals reaching for objects placed beyond the reach of the arm. Determining the control mechanism(s) governing this compensatory strategy in stroke patients was the goal of this study. Kinematics of reaching movements in hemiparetic and healthy participants to targets placed within and beyond the length of the arm were analysed. Targets were placed sagittally in front of the midline of the body. Two targets (targets 1 and 2) were within reaching distance defined as the length of the stretched arm from axilla to wrist crease. Two others were beyond arm's reach so that one required a forward trunk inclination (target 3) and the other required body raising to a semi-standing position (target 4). Healthy participants used minimal trunk displacement for reaches to targets 1 and 2. For reaches to targets 3 and 4, trunk displacement increased with target distance. Whenever the trunk was involved, there was a stereotyped sequential recruitment of the arm and trunk in that the trunk began moving simultaneously with or before the hand and stopped moving after the end of hand movement. This suggested that the control system predicts that the trunk movement will be needed to extend the reach and includes the trunk, in an anticipatory way, into the reach. In contrast, most hemiparetic participants recruited their trunk for reaches to all four targets, even those placed close to the body. Similar to healthy individuals, the sequence of hand and trunk recruitment was stereotyped, suggesting that temporal planning aspects of the motor program underlying movement coordination were relatively unaffected. In contrast to healthy participants, the contribution of the trunk movement to the endpoint displacement was substantially higher in the hemiparetic group and occurred earlier in the reach. It is suggested that the target distance at which the trunk is integrated into the movement to extend the reach of the arm is attained around the limit of arm extension and that this limit is reduced in hemiparetic individuals. PMID- 11880894 TI - How the lack of visuomotor feedback affects even the early stages of goal directed pointing movements. AB - Pointing movements made with a hidden cursor from the center of gaze to a stationary, visible target overshot the actual target location. The systematic error decreased when the final cursor location from the previous trial was shown, which likely led to the creation of an internal sensorimotor model of movement. However, the putative model had a short memory, and could not substitute for on line visuomotor feedback on subsequent trials. Contrary to common belief, the effect of a lack of visuomotor feedback was seen even in the early acceleration stage of the movement trajectory. Unchecked in the absence of visual monitoring, the acceleration stage of the movement lasted longer, as was evidenced by the significantly larger value of the peak cursor speed. Moreover, the speed peaked much later in the course of the movement. Speed declined more rapidly thereafter. Consequently, the delayed deceleration stage lasted far less than the acceleration stage. In the absence of visual feedback, the shift rightward in time of the peak speed position (PSP) in relation to total movement duration and other changes in the trajectory imply that visual feedback must play a significant role in determining when acceleration ceases (d V/d t=0), and argue against the traditional notion that visuomotor feedback is unavailable until the later stages of movement. Moreover, our data suggest that non-visual modalities, e.g., proprioception, may be too slow to make up for the absence of vision. PMID- 11880895 TI - Forced swimming test and fluoxetine treatment: in vivo evidence that peripheral 5 HT in rat platelet-rich plasma mirrors cerebral extracellular 5-HT levels, whilst 5-HT in isolated platelets mirrors neuronal 5-HT changes. AB - Low levels of central serotonin (5-HT) have been related to the state of depression, and 5-HT is the major target of the newer antidepressant drugs such as selective serotonin reuptake inhibitors (SSRIs). Neurons and platelets display structural and functional similarities, so that the latter have been proposed as a peripheral model of central functions. In particular, in blood more than 99% of 5-HT is contained in platelets, so that one could consider changes in 5-HT levels in platelets as a mirror of changes in central 5-HT. Here, this hypothesis has been studied via the analysis of the influence of: (1) the forced swimming test (FST, which has been proved to be of utility to predict the clinical efficacy of antidepressants in rodents) and (2) treatment with the SSRI fluoxetine upon 5-HT levels monitored in brain regions and in peripheral platelets by means of electrochemical in vivo and ex vivo measurements. The results obtained confirm that the FST increases immobility; furthermore they show a parallel and significant decrease in cerebral (brain homogenate) and peripheral (in platelet rich plasma, PRP) voltammetric 5-HT levels following the FST in naive rats. In addition, subchronic treatment with fluoxetine was followed by a significant increase in 5-HT levels in PRP, while the same SSRI treatment performed within the FST resulted in a decrease in the 5-HT levels in PRP. However, this decrease was inferior to that observed without SSRI treatment. These data suggest that there is an inverse relationship between immobility and the levels of 5-HT in PRP and that these peripheral 5-HT levels are sensitive to: (1) the FST, (2) the treatment with fluoxetine and (3) the combination of both treatments, i.e. SSRI + FST. It has been reported that SSRI treatment at first inhibits the 5-HT transporter in brain, resulting in increased extracellular 5-HT, while following sustained SSRI treatments decreased intracellular levels of central 5-HT were observed. Accordingly, the present data show that the initial block of 5-HT reuptake is revealed by the selective increase in 5-HT levels (extracellular content) measured in PRP (not in insulated platelets, IPs) the 1st day of fluoxetine treatment. The initial action of this SSRI upon the 5-HT transporter in brain has also been confirmed by in vivo voltammetric data showing selective increase in the serotonergic signal following local injection of fluoxetine into the brain region studied. Successively, the major effect monitored is a decrease in 5-HT levels, which is more evident in IPs than in PRP. However, it is known that following 2 weeks treatment with an SSRI, 5-HT autoreceptors are desensitized and the serotonin synthesis is restored, together with the intracellular 5-HT levels. The present data showing that the levels of 5-HT in IPs tend to return to control values 12 days after the beginning of chronic fluoxetine treatment suggest that 5-HT levels in IPs (intracellular environment) mirror the influence of SSRI treatment upon the central 5-HT system. On the other hand, at day 12 of the chronic fluoxetine treatment, 5-HT content remains low in PRP. Similarly, low levels of 5-HT have been monitored in brain homogenate of rats chronically treated with fluoxetine. This would support the similarity between PRP preparation and brain homogenate as in both cases cells are disrupted by sample preparation. In conclusion this work supports the literature in proposing platelets as a peripheral model of central functions. In particular, the present data support the idea that peripheral 5-HT platelet levels can reflect the state of the central 5-HT system in conditions of depression. Furthermore, the main outcome of this study is that PRP may mirror central extracellular 5-HT levels, whilst IPs mirror neuronal 5-HT changes. PMID- 11880896 TI - Hitting moving objects: is target speed used in guiding the hand? AB - We investigated what information subjects use when trying to hit moving targets. In particular, whether only visual information about the target's position is used to guide the hand to the place of interception or also information about its speed. Subjects hit targets that moved at different constant speeds and disappeared from view after varying amounts of time. This prevented the subjects from updating position information during the time that the target was invisible. Subjects hit further ahead of the disappearing point when the target moved faster, but not as much as they should have on the basis of the target's speed. This could be because more time is needed to perceive and use the correct speed than was available before the target disappeared. It could also be due to a speed related misperception of the target's final position. The results of a second experiment were more consistent with the latter hypothesis. In a third experiment we moved the background to manipulate the perceived speed. This did not affect the hitting positions. We conclude that subjects respond only to the changing target position. Target speed influences the direction in which the hand moves indirectly, possibly via a speed-related misperception of position. PMID- 11880897 TI - Force adaptation transfers to untrained workspace regions in children: evidence for developing inverse dynamic motor models. AB - When humans perform goal-directed arm movements under the influence of an external damping force, they learn to adapt to these external dynamics. After removal of the external force field, they reveal kinematic aftereffects that are indicative of a neural controller that still compensates the no longer existing force. Such behavior suggests that the adult human nervous system uses a neural representation of inverse arm dynamics to control upper-extremity motion. Central to the notion of an inverse dynamic model (IDM) is that learning generalizes. Consequently, aftereffects should be observable even in untrained workspace regions. Adults have shown such behavior, but the ontogenetic development of this process remains unclear. This study examines the adaptive behavior of children and investigates whether learning a force field in one hemifield of the right arm workspace has an effect on force adaptation in the other hemifield. Thirty children (aged 6-10 years) and ten adults performed 30 degrees elbow flexion movements under two conditions of external damping (negative and null). We found that learning to compensate an external damping force transferred to the opposite hemifield, which indicates that a model of the limb dynamics rather than an association of visited space and experienced force was acquired. Aftereffects were more pronounced in the younger children and readaptation to a null-force condition was prolonged. This finding is consistent with the view that IDMs in children are imprecise neural representations of the actual arm dynamics. It indicates that the acquisition of IDMs is a developmental achievement and that the human motor system is inherently flexible enough to adapt to any novel force within the limits of the organism's biomechanics. PMID- 11880898 TI - "Look where you're going!": gaze behaviour associated with maintaining and changing the direction of locomotion. AB - In order to fully understand how vision is used to guide locomotion it is necessary to know what people look at as they move through the environment. This study provides information, hitherto lacking, regarding gaze behaviour associated with both maintaining and changing the direction of locomotion: activities that are essential for efficient navigation through our cluttered environment. Participants' spatiotemporal gaze patterns were recorded whilst they performed a task requiring that they either maintained a straight walking trajectory or changed their direction of walking by 30 degrees or 60 degrees, left or right, at the midpoint of a 9-m path. Participants were either visually cued to turn when they stepped on a trigger mat placed one step before the mid-point of the walkway (cued trials) or given verbal instruction about the required route prior to the start of each trial (advance knowledge trials). Our clear finding was that for the large majority of the time participants' gaze was aligned with environmental features lying in their current plane of progression both prior to and following the onset of the transition stride during which the direction change was implemented. This gaze behaviour was observed both during cued trials (78% of total fixation time prior to the transition stride onset and 89% following the transition stride onset) and advance knowledge trials (67% prior to transition stride onset, 92% following transition stride onset). When not aligned with the plane of progression, gaze was normally fixated on environmental features related to either known or potential future routes. Prior to changing the direction of walking, individuals invariably made saccadic eye movements in order to align gaze with the end-point of the required travel path. This gaze realignment was invariably accompanied by head reorientation, which was initiated, on average, at the same time as the saccade. On average, participants fixated gaze on their goal (represented by the cue light at the travel path end-point) until after head realignment with the new path was achieved. Additionally, the head was consistently aligned with participants' current walking direction prior to and following the transition stride even on the minority of occasions when they were looking elsewhere. These findings challenge the ecological validity of existing theories of how visual information is used to determine heading direction and are consistent with the proposal that aligning the head with the desired travel direction through coordinated eye and head movements provides the CNS with an allocentric frame of reference that is used to control the movement of the body in space. PMID- 11880899 TI - Effects of tendon vibration on the spatiotemporal characteristics of human locomotion. AB - The present study addressed the involvement of proprioceptive input of the muscle spindles in the spatiotemporal control of human locomotion. Blindfolded subjects walked along a walkway while tendon vibration, a powerful stimulus of Ia afferents, was applied to various muscles of the lower limb. The effects of tendon vibration were measured on joint kinematics and on intralimb and interlimb coordination. Tendon vibration of the tibialis anterior during locomotion led to a decreased plantar flexion at toe-off, whereas vibration of the triceps surae led to a decreased dorsiflexion during swing. Vibration of the quadriceps femoris at the knee led to a decreased knee flexion during swing. These local effects of vibration can be explained in the light of a lengthening illusion of the vibrated muscle in that phase of the gait cycle where the muscle is lengthened. Tendon vibration did not affect the qualitative features of intralimb coordination. With respect to interlimb coordination, only vibration of the biceps femoris showed a significant increase in phase lead of the vibrated limb. The present results suggest the involvement of Ia afferent input in the online control of joint rotations. Additionally it is hypothesized that the proprioceptive input of biceps femoris might be involved in the control of coordination between the limbs, whereas the coordination between the segments of one limb appears to be unaffected by disturbance of muscle spindle input of one muscle. PMID- 11880900 TI - Two phases of intracortical inhibition revealed by transcranial magnetic threshold tracking. AB - Intracortical inhibition was investigated in normal human volunteers by paired pulse transcranial magnetic stimulation, using a new, computer-assisted threshold tracking method. Motor threshold was defined as the stimulus amplitude required to evoke a motor evoked potential of 0.2 mV (peak-to-peak) in abductor pollicis brevis, and inhibition was measured as the percentage increase in threshold, when the test stimulus was preceded by a subthreshold conditioning stimulus. This method was used to investigate the dependence of intracortical inhibition on conditioning stimulus parameters and on voluntary activity. Interstimulus interval (ISI) was first stepped from 1 to 4.5 ms, as inhibition was measured using conditioning stimuli of fixed amplitude (50-90% resting motor threshold). Maximal inhibition was produced at ISIs of 1 and 2.5 ms. The effect of conditioning stimulus intensity was then assessed at these ISIs. Inhibition occurred at significantly lower conditioning stimulus intensities with ISI=1 ms than with ISI=2.5 ms. Voluntary activity reduced inhibition at both ISIs, but had a much greater effect on inhibition at ISI=2.5 ms. Inhibition during voluntary activity was also examined for single motor units in first dorsal interosseous by generating poststimulus time histograms. Inhibition, indicated by a reduction in the later peaks of increased firing, was observed with ISI=1 ms, but not with ISI=2.5 ms. We conclude that there are two distinct phases of inhibition, occurring at ISI=1 ms and ISI=2.5 ms, differing both in thresholds and susceptibility to voluntary activity. PMID- 11880901 TI - Psychophysical and pupillometric study of spatial channels of visual processing in blindsight. AB - To date no systematic method has been used for characterising the residual capacity of blindsight subjects that would allow comparison and generalisation across all subjects. The detection of isoluminant gratings of varying spatial and temporal frequencies commends itself for detailed between-subject comparison, and for mapping results onto physiological properties in relation to neuronal circuitry. We report the ability of a blindsight subject (CS) to detect suprathreshold sine-wave gratings over a range of spatial and temporal frequencies using psychophysical techniques. A band-pass spatial channel with an upper cutoff below 3.5 cycles/deg is specified. The data also have been analysed to compare differences between two types of blindsight performances, type I and type II. Spatial gratings were also used to elicit a pupillary grating response, offering an objective method that is free of verbal nuances and response bias, and the resulting band-pass channel can be used both for clinical screening and for prediction and comparisons with psychophysical profiles. Finally, we have compared our results with those reported in studies of a well-known subject, GY, which demonstrate remarkable similarities. Implications are discussed in relation to blindsight research. PMID- 11880903 TI - Distribution-dependent saccades in children with strabismus and in normals. AB - To guide saccades accurately human adult subjects learn rapidly to aim, in advance, for the central position of the distribution of the target positions studied. This causes a bias called range effect, i.e., saccades to the central location are accurate while the less eccentric locations are overshot and the more eccentric locations are undershot. It is believed that this is a cognitive strategy allowing to reduce overall variability. We show here that the same exists for normal children (5-13 years old). Most importantly, from the age of 5 years, children with strabismus use the same cognitive strategy despite the fact that their eyes are misaligned. Importantly, both eyes follow this strategy even though only one eye is fixating the target (the other eye being squinted). PMID- 11880902 TI - Depletion of intracellular glutathione mediates zinc-induced cell death in rat primary astrocytes. AB - In the present study, we investigated the possible mechanisms of cellular injury induced by zinc in rat primary astrocytes and C6 glioma cells. Reactive oxygen species (ROS) production, cellular glutathione (GSH) level and mitochondrial transmembrane potential were examined. Exposure to 200-300 microM Zn2+ for 24 h resulted in significant lactate dehydrogenase (LDH) release in rat primary astrocytes and C6 glioma cells. An exposure of 200 microM Zn2+ resulted in profound morphological changes, for example, shrunken and fragmented nuclei. Pretreatment of a protein synthesis inhibitor, cycloheximide, did not attenuate cellular toxicity induced by Zn2+. Zn2+ exposure increased intracellular ROS levels by about 250%, and depleted cellular GSH within 2 h, which preceded observable LDH release from the cell. Addition of GSH, N-acetylcysteine (NAC) and ascorbic acid substantially attenuated cellular death induced by Zn+ in a concentration dependent manner. ROS production and morphological changes induced by zinc were also inhibited by co-treatment of GSH or NAC with Zn2+. Zn2+ significantly depolarized mitochondrial transmembrane potential, which was reversed by co-treatment of GSH or NAC with zinc. In summary, ROS generation, GSH depletion and mitochondrial dysfunction may be key factors in Zn2+-induced glial toxicity. PMID- 11880904 TI - Subvastus versus medial parapatellar approach in total knee arthroplasty. AB - The subvastus approach for total knee replacement was compared with the standard medial parapatellar approach in terms of postoperative knee scores and quadriceps strength. Two groups of patients with similar characteristics were formed: the first group consisted of 12 knees of 9 patients who were implanted via the medial parapatellar approach, and for the second group the subvastus approach was used in 10 knees of 10 patients. The groups' knee scores and quadriceps strength were compared preoperatively and postoperatively at week 6, months 3 and 6. The knee scores improved similarly in both groups, but the change was more pronounced in the subvastus group. Quadriceps strength was greater in the subvastus group at postoperative week 6, but there was no significant difference between the groups in months 3 and 6. It was concluded that although the subvastus approach offers greater quadriceps strength in the early postoperative period, it has no significant advantage in this aspect over the medial parapatellar approach. PMID- 11880905 TI - Basicervical fracture--a rare type of hip fracture. AB - Basicervical fracture is a controversial type of hip fracture, which can be regarded as either extracapsular or intracapsular. It is seldom mentioned in the authorized orthopaedic textbooks, and it lacks an exact definition in the most commonly used classifications. The aim of this study was to evaluate the rate of basicervical hip fractures and the methods of treating them in a prospective series of 1624 consecutive hip fractures. Standardized forms were used to collect information, including the classification of fracture types. Initially, 108 fractures were classified as basicervical, but a careful second-look check revealed that 51 were transcervical fractures, while 27 fractures had a trochanteric extension. Thus, 30 of the fractures fulfilled the criteria of basicervical fracture (rate 1.8%). The 14 fractures treated as extracapsular fractures (dynamic hip screw, DHS, or gamma nail) showed a better outcome than the 16 treated as intracapsular fractures (hemiarthroplasty or screw osteosynthesis). We conclude that basicervical fracture of the hip is a very uncommon entity, but it is worth considering and should be treated as a trochanteric fracture. PMID- 11880906 TI - Problems of posterior lumbar interbody fusion (PLIF) for the rheumatoid spondylitis of the lumbar spine. AB - We performed posterior lumbar interbody fusion (PLIF) on 7 patients with rheumatoid spondylitis (RA) of the lumbar spine with severe low-back pain and/or cauda equina symptoms, and evaluated the effectiveness of PLIF for the lumbar spinal instability in RA secondary to destruction of the anterior elements, including vertebral endplates and the apophyseal joint. The subjects were 7 patients with classic RA, 2 men and 5 women, mean age 65 years old, and the mean duration of RA was 21 years. All had severe low-back pain and difficulty with walking. According to the ARA classification, the patients were at stage 3 or worse and in class 3. Diagnostic imaging including magnetic resonance imaging (MRI), tomography, myelography, and computed tomographic myelography (CTM) of the lumbar spine clearly delineated pathology, destruction of the vertebral endplate, subluxation, and cauda compression which can be well treated with PLIF. We performed L4/5PLIF (5 cases), L3/4 and L4/5 PLIF (2 cases), and posterior fixation with instruments for anterior column repair and stabilization and posterior decompression. Autografts (all cases) and Brantigan IF cage (2 cases) were used. Stable fixation of the lumbar spine was achieved after surgery, and improvement in gait and activities of daily living were achieved through the relief of low-back pain and radicular pain; the mean duration of follow-up was 22 months. Postoperative, plain radiography, CT, and MRI revealed the enlargement of the lumbar canal and fusion and incorporation of grafted bone, but in some cases, collapse of graft, migration of pedicle screw, instability of adjacent level, and collapse of adjacent vertebra were noted. PLIF with spinal instruments is a preferred treatment for rheumatoid spondylitis of the lumbar spine, but in the mutilating type of RA with severe osteoporosis, PLIF in combination with a long fixation system and/or augmentation of the vertebral bodies might be needed. PMID- 11880907 TI - Successful hip arthroplasty using cementless titanium implants in rheumatoid arthritis. AB - Over a period of eight years, we implanted a total of 76 cementless hip prostheses in patients with rheumatoid arthritis. The clinical results of 47 patients (70 hips) increased from a mean Harris Hip Score of 33 to 85 after an average of 49 months (range 1-11 years). One threaded cup has had to be revised because of loosening, and one stem because of femoral fracture. At the latest follow-up, 88% of Hofer-Imhof threaded cups had complete bone ingrowth (Type 0); 10% had near-complete bone ingrowth with minimal radiolucency in one third of the bone contact area (Type 1), and 2% had radiolucency in two thirds of the bone contact area (Type 2). Hemispherical push-in cups showed significantly more radiolucency around the cup. For the stems (Uni, Zweymuller SL), 83% showed no radiolucency (Type 0); 17% had radiolucency only very proximally (Type 1). Minor remodelling (Type 1) occurred in 60% of the femoral shafts; 30% had moderate femoral density loss (Type 2), and 10% had severe bone loss and cortical thinning (Type 3). There was no correlation between marked shaft atrophy and clinical symptoms. With regard to radiolucency and remodelling, there was no significant difference between the two types of stem used. Cementless hip arthroplasty using titanium implants has an excellent outcome in the medium term. PMID- 11880908 TI - Mechanics of femoral head osteonecrosis using three-dimensional finite element method. AB - A three-dimensional finite element model of a femoral head was developed using a surface modeling technique. The distribution of the stress index S (S = effective stress / yield strength, sigma/sigmaY) in various sizes of segmental osteonecrosis was assessed. The stress index of the femoral head was within physiological limits when the necrotic angle was less than 110 degrees. Within both the subchondral region and the deep necrotic region adjacent to the necrotic viable interface, values of the stress index significantly higher than the normal physiological level (>0.1) appeared when the necrotic angle was 110 degrees or more. In the analysis of 28 osteonecrotic femoral head specimens, fracture appeared in two major locations: the deep necrotic region near the underlying necrotic-viable interface (19 femoral heads) and the subchondral region (7 femoral heads). In 2 femoral heads, the fracture involved both regions. Both sites of fracture coincided with the region of stress index greater than 0.1 in the finite element model study. These results may provide baseline information for predicting the collapse of the femoral head and determining the treatment modality of early stage osteonecrosis. PMID- 11880909 TI - Three-dimensional virtual computed tomography imaging for injured anterior cruciate ligament. AB - Three-dimensional virtual imaging using reconstructed volume data from helical computed tomography was applied to examine injured anterior cruciate ligaments (ACL) as a noninvasive diagnostic aid. Its diagnostic value for ACL injury was evaluated in 21 patients. These results were compared with those of arthroscopic findings. This new technique allowed visualization of the ACL in a three dimensional form, similar to an arthroscopic image, and was 100% accurate in detecting ACL injuries in the present samples. PMID- 11880910 TI - Effect of continuous versus dichotomous outcome variables on study power when sample sizes of orthopaedic randomized trials are small. AB - It is often not feasible to conduct large trials in orthopaedic surgery. Therefore, surgeons must identify strategies to optimize the statistical power of their smaller studies. The aim of this study was to compare study power in randomized trials with continuous versus dichotomous outcome variables. We performed a systematic review of the literature to identify randomized trials in orthopaedic trauma. Of these, we examined only those trials with small sample sizes (50 patients or less). The outcomes in each eligible study were categorized as continuous or dichotomous. Standard power calculations were performed for each study, and comparisons were made between continuous and dichotomous outcome variables. We identified 196 randomized trials in orthopaedic trauma. Of these, 76 trials had a sample size of 50 patients or fewer (29 trials with continuous outcomes, 47 trials with dichotomous outcomes). Studies that reported continuous outcomes had a significantly higher mean power than those that reported dichotomous variables (power 49% vs 38%, p=0.042). Twice as many trials with continuous outcome variables reached acceptable levels of study power (i.e. >80% power) when compared with trials with dichotomous variables (37% vs 18.6%, p=0.04). When orthopaedic surgeons anticipate small sample sizes for their study, they can optimize their study's statistical power by choosing a continuous outcome variable. PMID- 11880911 TI - Normal range of scapular elevation and depression in healthy subjects. AB - The scapular elevation (shoulder shrug), depression, and total elevation depression motion in 30 healthy subjects were measured by goniometer under standardized conditions. Mean values were 37.74, 8.39, and 44.60 deg, respectively. The intraobserver variation was calculated as 1.3 deg, and the interobserver variation was 1.8 deg. There was no difference between the sides. This previously unreported values may be used in the evaluation of shoulder and neck disorders. PMID- 11880912 TI - Functional treatment after surgical repair of acute Achilles tendon rupture: wrap vs walking cast. AB - The aim of this study was to compare the clinical outcome for patients treated with walking cast immobilization and wrap early mobilization after surgical repair of acute Achilles tendon ruptures. A total of 39 consecutive patients with complete ruptures of the Achilles tendon were identified, treated, and functionally rehabilitated with either a walking cast or a wrap. Because the randomization was quasi-random, chi-square and t-tests were performed to compare the baseline characteristics. A statistically significant difference was present only for the injured side ( p<0.05). Therefore, groups were considered comparable for analysis of outcome. All patients were evaluated at an average follow-up of 6.7 years (range 5-8 years). Functional postoperative treatment with a wrap allowed a significantly shorter hospital stay ( p<0.05) as well as a shorter period to return to pre-injury sports level ( p<0.01) compared with treatment with a walking cast. According to the modified Rupp score, 91.3% of patients in the walking cast group had a good or excellent result, as did 93.8% in the wrap group ( p=0.9). Slight atrophy of the calf muscles was reported in 3 patients in the walking cast group (13.0%) and in 4 in the wrap group (25.0%). One re-rupture was reported in the walking cast group (4.3%). Functional treatment after surgical Achilles tendon repair is safe, and there is no increased risk of re rupture or wound healing problems. Functional treatment with a wrap is preferable to treatment with a walking cast with respect to hospitalization time and return to sports. PMID- 11880914 TI - Spontaneous haematoma of the iliac psoas muscle: a case report and review of the literature. AB - Spontaneous haematomas of the iliac psoas muscle are rare lesions seen in patients receiving anticoagulant agents or suffering from clotting disorders. We report the rare case of a spontaneous iliac psoas haematoma causing femoral neuropathy in a patient not undergoing anticoagulant treatment or suffering from a coagulopathy. After conservative treatment the patient was entirely asymptomatic, and magnetic resonance imaging demonstrated complete dissolution of the haematoma. Spontaneous iliac psoas haematomas should be considered in the differential diagnosis of leg pain in otherwise healthy patients. PMID- 11880913 TI - Metastatic spinal neurofibrosarcoma. AB - Neurofibrosarcomas are rare tumors usually arising in somatic soft tissues or peripheral nerves. Four cases of metastatic neurofibrosarcoma to the spine have been reported before. The current case is unusual because of the presence of two distinct, metachronous spinal metastasis and lung metastasis. A 30-year-old woman with neurofibromatosis and a history of previous neurofibrosarcoma resection presented with back pain. Radiologic evaluation revealed a lytic lesion of the eleventh thoracic vertebra. A transthoracal corpectomy, reconstruction by Harms' cage and posterior instrumentation, and fusion were carried out. After the completion of adjuvant chemotherapy, a solitary pulmonary nodule was detected. Shortly after resection of the metastatic pulmonary nodule, the patient complained of acute onset low-back pain. Radiologic assessment revealed another lytic lesion in the L5 vertebra after 6 months. Again, a corpectomy, anterior and posterior instrumentation, and fusion were carried out. Eight months after the second spinal resection, another solitary pulmonary metastasis was diagnosed and resected. The patient's health status suddenly deteriorated 26 months after the initial spinal metastatectomy, and she died. Though local control can be achieved in more than 80% of the patients with neurofibrosarcoma by wide surgical resection followed by adjuvant chemo- and radiotherapy, most patients die of systemic metastasis. The current patient survived 50 months after the initial resection of a forearm neurofibrosarcoma. Despite achieving local control, she died due to systemic recurrence. Prolonged survival with the help of chemo- and radiotherapy justifies our aggressive surgical strategy for the treatment of spinal metastasis in order to achieve neurologic cure and spinal stability. PMID- 11880915 TI - Multifocal soft-tissue metastases of gastric leiomyosarcoma. AB - Distant soft-tissue metastases in gastric leiomyosarcoma (LMS) are extremely rare. In a 65-year-old woman, a large, low-grade LMS originating from the stomach wall was treated by partial gastrectomy. Eight years later, the patient developed multifocal, soft-tissue, high-grade LMS. These tumors were presumably metastases of the gastric LMS. However, due to the demonstration of mutations in some cases of leiomyosarcoma, the possibility of multifocal sarcoma should be kept in mind. PMID- 11880916 TI - Hyperbaric oxygenation as a successful therapeutic approach in oral wound dehiscence after operative stabilization of an unstable post-traumatic odontoid non-union. AB - The non-operative treatment of unstable traumatic Anderson's type II odontoid fractures has a high risk potential to develop non-unions. Even after operative stabilization literature reveals non-union rates up to 20%. Acute life threatening complications are tetraplegia and apnoea. Long-term complications induce chronic myelopathy resulting from persistent myeloradicular compression. We report the case of a patient with a 17-year-old post-traumatic pseudarthrosis of the dens axis following conservative treatment of an unstable type II fracture. By that time, the female patient, then 37 years old, was admitted to our hospital with early signs of cervical tetraplegia. After initial reposition and short-term immobilization with a halothoracic vest we performed a ventrodorsal atlantoaxial spondylodesis. Failure of anterior cervical plate stabilization and autologous graft resorption without a solid segmental fusion instigated a secondary surgical intervention. Postoperative therapy-resistant oral wound dehiscence showed an exposed autograft and osteosynthetic material. The reported positive effect of hyperbaric oxygenation on wound healing in problem cases led us to attempt this means of therapy. With a daily exposure to hyperbaric oxygenation, the dehiscence closed within 25 days. As a result of our experience in this case, hyperbaric oxygenation should be considered as a therapeutic option in postoperative complication management in orthopaedic surgery. PMID- 11880917 TI - Myelopathy due to scoliosis with vertebral hypertrophy in Klippel-Trenaunay-Weber syndrome. AB - We describe a 37-year-old man complaining of right back pain and gait disturbance. He had a big soft tumor on his right back, hemihypertrophy of the right lower extremity, and right thoracic scoliosis. We diagnosed Klippel Trenaunay-Weber syndrome based on the pathological findings of the soft tumor. Computed tomography (CT) scan revealed severe spinal stenosis due to a hypertrophic vertebral body and facet joint at T7. Treatment by decompression of hypertrophic bone led to complete neurological recovery. To our knowledge, no case has been reported of Klippel-Trenaunay-Weber syndrome with myelopathy which originated from thoracic scoliosis with a hypertrophic facet joint and vertebral body. We suggest that the cause of myelopathy in Klippel-Trenaunay-Weber syndrome originated not only from arteriovenous fistula, medullary angioma, and extradural hemangioma but also vertebral hypertrophy with scoliosis. PMID- 11880919 TI - Introduction to this special issue on diabetes mellitus and its complications. PMID- 11880918 TI - Knee arthrodesis with a temporary spacer performed in malignant tumor around the knee. AB - The issue of utilizing a temporary cement spacer after a knee resection arthrodesis of a highly malignant bone tumor is discussed. In our experience, this is the first type of reconstruction performed in the early days of the limb salvage. Now, after more than 12 years of follow-up based on the results of 84 patients, the usefulness and mechanical performance of this type of reconstruction are considered. Of these patients, 45 died of disease. Wound complications were present in 31% of the patients and led to amputation in 7 and healing failure in 6. Mechanical complications occurred in 39%, but radiographic analysis of the implant showed 32 cases (38%) with an inadequate spacer. Only 9 patients are still walking on the original reconstruction, 6 of them uncomplicated (the remaining 3 had rod migrations and shortening). The survival curve showed no statistical difference between the two groups (adequate vs inadequate spacer). In 33 patients who are still alive and retain their limb, the final functional results were excellent in 6, good in 8, fair in 14, and poor in 5. This type of reconstruction can still be considered in developing countries, or when a definitive arthrodesis is planned where there is a lack of a bone source (children, no bone bank). In our institution, reconstruction with cement is still suggested for patients with a large tumor around the knee, when the quadriceps is completely sacrificed, and the patient prognosis is markedly unfavorable. PMID- 11880920 TI - Diabetes mellitus - multifactorial in aetiology and global in prevalence. PMID- 11880921 TI - Characterisation of ventricular myocyte shortening after administration of streptozotocin (STZ) to neonatal rats. AB - Human and animal studies have shown that diabetes mellitus can be associated with altered cardiac function that is independent of vascular complications. Streptozotocin (STZ) is a diabetogenic agent and when administered to rats causes selective beta-cell necrosis which is accompanied by a drastic reduction in plasma insulin and hyperglycaemia. We have investigated the characteristics of shortening in ventricular myocytes isolated from rat heart at 10 months after administration of STZ to neonatal rats at 2 days of age. The characteristics of shortening in myocytes from the STZ-treated neonatal rat compared to shortening in myocytes from the STZ-treated young adult rat are discussed. STZ-treated rats gained significantly less weight compared to age-matched controls. Although non fasting blood glucose was not significantly different in STZ-treated rats they were found to be markedly glucose intolerant when given an intraperitoneal challenge of glucose (2 g/kg) after an overnight fast. During electrical stimulation (1 or 2 Hz) ventricular myocyte resting length, time to peak shortening, time to half relaxation and amplitude of shortening were not altered after STZ treatment. The imposition of rest periods (2-60 s) after trains of electrically stimulated (1 Hz) steady-state contractions resulted in a potentiation of the contraction, which immediately followed the rest period (post rest potentiation). Post rest potentiation was larger, following rest periods between 20 and 60 s, in myocytes from STZ-treated rats compared to controls. The absence of major alterations to the amplitude and kinetics of contraction in myocytes from STZ-treated neonatal rats at 10 months after treatment might be explained by a partial recovery of the beta-cells in the growing animal. PMID- 11880922 TI - Catecholamines in the heart and adrenal gland of the STZ-diabetic rat. AB - The concentrations of noradrenaline (NA), adrenaline (ADR), 5-hydroxyindoleacetic acid (5-HIAA), serotonin (5-HT) and dopamine (DOP) have been studied in the left ventricle and the left adrenal gland of control and streptozotocin (STZ) - treated rats at various intervals (12, 24, 30, 34, 38 and 42 weeks) after the induction of diabetes. The only amines detected in the heart were NA, 5-HIAA and DOP, whereas those detected in the adrenal gland were NA and ADR. Differential changes in the catecholamine concentrations occurred in the heart and the adrenal gland at different stages of the metabolic disorder. In the heart the initial changes in short-term diabetes included an increase in NA concentration but this did not persist in the longer term diabetic animals (30-38 weeks following STZ injection). In the adrenal gland there was an initial reduction followed by a steady increase in the concentration of NA and ADR throughout the period of the study. PMID- 11880923 TI - Abnormal vascular coiling of the umbilical cord in gestational diabetes mellitus. AB - The objective of this study was to identify abnormal vascular coiling of the umbilical cord in neonates of mothers with gestational diabetes mellitus. The umbilical cords of 57 neonates of gestational diabetic mothers were examined and the coiling index determined by dividing the total number of complete vascular coils by the length of the cord in centimeters. Obstetric history, delivery data and neonatal outcome were also evaluated. These variables were compared with those obtained for 389 normal pregnancies. The frequency distribution of umbilical coiling index in the control population and gestational diabetic mothers were normal (10th and 90th percentiles = 0.17 and 0.37; mean +/- SD = 0.26 +/- 0.09 and 0.24 +/- 0.12 coils/cm, respectively). Non-coiling and hyper coiling were significantly more frequent with diabetic than with normal pregnancy (p = 0.004; p = 0.008, respectively). Both abnormalities of umbilical vascular coiling were also significantly associated with adverse perinatal outcome (p = 0.04) and emergency cesarean delivery (p < 0.0001) in the diabetic and control (p = 0.03; p < 0.0001, respectively) groups. Neonates of gestational diabetic mothers are therefore more likely to have hyper-coiled or non-coiled umbilical blood vessels. Perinatal morbidity and emergency cesarean delivery are increased in this subgroup. PMID- 11880924 TI - Blood pressure response to standing in the diagnosis of autonomic neuropathy: the EURODIAB IDDM Complications Study. AB - Autonomic neuropathy is associated with poor prognosis. Cardiovascular reflexes are essential for the diagnosis of autonomic nerve dysfunction. Blood pressure response to standing is the most simple test for the evaluation of sympathetic integrity, however it is still discussed which diagnostic criteria of abnormal response should be considered as optimal. The EURODIAB IDDM Complications Study involved the examination of randomly selected Type 1 diabetic patients from 31 centres in 16 European counties. Data from 3007 patients were available for the present evaluation. Two tests of autonomic function (response of heart rate /R-R ratio/ and blood pressure from lying to standing) just as the frequency of feeling faint on standing up were assessed. R-R ratio was abnormal in 24% of patients. According to different diagnostic criteria of abnormal BP response to standing (>30 mmHg, >20 mmHg, and >10 mmHg fall in systolic BP), the frequency of abnormal results was 5.9%, 18% and 32%, respectively (p < 0.001). The frequency of feeling faint on standing was 18%, thus, it was identical with the prevalence of abnormal blood pressure response to standing when >20 mmHg fall in systolic blood pressure was considered as abnormal. Feeling faint on standing correlated significantly with both autonomic test results (p < 0.001). A fall >20 mmHg in systolic blood pressure after standing up seems to be the most reliable criterion for the assessment of orthostatic hypotension in the diagnosis of autonomic neuropathy in patients with Type 1 diabetes mellitus. PMID- 11880925 TI - The role of leucine-enkephalin on insulin and glucagon secretion from pancreatic tissue fragments of normal and diabetic rats. AB - Leucine-enkephalin (Leu-Enk) has been shown to be present in endocrine cells of the rat pancreas and may play a role in the modulation of hormone secretion from the islets of Langerhans. Since little is known about the effect of Leu-Enk on insulin and glucagon secretion, it was the aim of this study to determine the role of Leu-Enk on insulin and glucagon secretion from the isolated pancreatic tissue fragments of normal and diabetic rats. Pancreatic tissue fragments of normal and streptozotocin-induced diabetic rats were incubated for 1 h with different concentrations of Leu-Enk (10(-12)-10(-6)M) alone or in combination with either atropine or yohimbine or naloxone. After the incubation period the supernatant was assayed for insulin and glucagon using radioimmunoassay techniques. Leu-Enk (10(-12 )-10(-6)M) evoked large and significant increases in insulin secretion from the pancreas of normal rats. This Leu-Enk-evoked insulin release was significantly (p < 0.05) blocked by atropine, naloxone and yohimbine (all at 10(-6)M). In the same way, Leu-Enk at concentrations of 10(-12)M and 10( 9)M induced significant (p < 0.05) increases in glucagon release from the pancreas of normal rats. Atropine, yohimbine but not naloxone significantly (p < 0.05) inhibited Leu-Enk-evoked glucagon release from normal rat pancreas. In contrast, Leu-Enk failed to significantly stimulate insulin and glucagon secretion from the pancreas of diabetic rats. In conclusion, Leu-Enk stimulates insulin and glucagon secretion from the pancreas of normal rat through the cholinergic, alpha-2 adrenergic and opioid receptor pathways. PMID- 11880926 TI - Neurogenic inflammation of gingivomucosal tissue in streptozotocin-diabetic rat. AB - Previously it was assumed that nerve fibres are involved in the neurogenic inflammation induced by mechanical or chemical irriations. It has been also suggested that in diabetes mellitus the unmyelinated small diameter fibers are impaired as a result of diabetic neuropathy. Therefore, our aim was to study the alterations of the nerve processes in the gingivomucosal tissue in streptozotocin (STZ)-diabetic rats. Light- and electronmicroscopical examinations were made to analyze the changes in nerve fibres. After one week of steptozotocin treatment, the gingivomucosal tissue had inflammatory cell infiltration and some degenerated nerve fibres were also observed. Dense mitochondria, disorganization of cell organelles, and appearance of myelin-like dense bodies were found in the axons of degenerared nerve fibres. Semiquantitative analysis showed that 14 +/- 4% of the unmyelinated nerve fibres degenerated after one week of STZ treatment. However, degeneration of the myelinated nerve fibers was not observed. Two weeks after STZ treatment, most of the unmyelinated and myelinated nerve fibers showed degeneration (86 +/- 5%) and the placement of the ligature revealed a non inflammatory connective tissue adjacent to a normal epithelium. The myelin sheath was disrupted and dark axoplasm with cytolysosomes became manifest. These findings demonstrated that both unmyelinated and myelinated nerve fibers are altered and inflammatory reaction exists in the gingivomucosal tissue only in the early stage of diabetes mellitus. PMID- 11880927 TI - Electrophysiological changes in diabetic neuropathy: from subclinical alterations to disabling abnormalities. AB - Clinical spectrum of diabetic neuropathy is variable; it may be asymptomatic, but once established as polyneuropathy, it is irreversible and may finally be disabling. To estimate the prevalence of subclinical diabetic polyneuropathy in the UAE, we undertook a pilot study by means of nerve conduction study (NCS) of peroneal motor and sural sensory studies in 60 diabetics with no symptoms of neuropathy. Neurological examination revealed clinical abnormalities suggesting polyneuropathy in 26 patients, 43% of the patients. NCS revealed abnormal values in 63% of the whole patients. Abnormal NCS was confirmed in 88% of the positive sign group. As to the negative sign group 44% had abnormalities in NCS. Prolonged F-wave latency was seen in 29% in no sign group and in 66% of the patients with positive signs. We found close association between neurological deficit score and abnormalities in NCS. Among various parameter of systemic nerve conduction study in subclinical patients, prolonged F-wave latency seems the commonest abnormality suggesting morphological changes in subclinical diabetic nerve. Decrease in amplitude of compound sensory action potential of sural nerve is another earlier abnormality, which is, then, accompanied by a fall in motor amplitude of peroneal nerve in advanced patients. Recently, our own group of Hirosaki has demonstrated that somatosensory central conduction time (CCT) between the spinal cord entry time and the arrival time to the sensory cortex is prolonged in diabetics. This abnormality might be partly responsible for the irreversible sensory deficits of diabetic neuropathy. PMID- 11880928 TI - Morphological changes in the rat kidney following long-term diabetes. AB - The morphological basis of diabetic nephropathy has been studied using light and electron microscopy. Kidneys of streptozotocin-induced diabetic rats were examined on the light microscope at 4 weeks and 8 months after induction of diabetes mellitus. In addition, the 8-month diabetic kidneys were examined with the electron microscope. Renal hypertrophy was evidenced by the increase in the weight of kidneys of diabetic rats. Whilst the diabetic kidneys were approximately twice as large after 4 weeks they were only 30% larger compared to age-matched controls after 8 months of induction of diabetes. After 4 weeks, light microscopy revealed dilated tubules within the cortex of the diabetic kidneys. Light microscopy showed a significant amount of destruction of the distal convoluted tubules while electron microscopy revealed a spectrum of damage that included basement membrane thickening, loss of podocytic foot processes, disruption of tubular basal infoldings and their related mitochondria and fibrosis of the tubules 8 months after induction of diabetes. It is concluded that renal hypertrophy persists after a prolonged occurrence of diabetes but the extensive damage and loss of renal tissue including the loss of the foot processes of podocytes might be partly responsible for the clinical presentation of diabetic nephropathy. PMID- 11880929 TI - Diabetes mellitus is associated with a decrease in vasoactive intestinal polypeptide content of gastrointestinal tract of rat. AB - Vasoactive intestinal polypeptide (VIP) is an inhibitory non-adrenergic, non cholinergic transmitter, which mediate in the relaxation of sphincters of the gastrointestinal tract. The aim of this study was to determine whether there is a change in the pattern of innervation and tissue content of VIP in the rat gastroduodenum after the onset of streptozotocin (STZ)-induced diabetes mellitus. Diabetes was induced by a single intraperitoneal injection of STZ (60 mg Kg(-1)). Four weeks after the induction of diabetes mellitus, the rats were anaethetised and the pancreata were removed for further processing. VIP was localized and measured in normal and diabetic rat gastroduodenal tissues by immunohistochemistry and radioimmunoassay, respectively. VIP immunoreactivity was stronger in the ganglion cells of the submucosal and myenteric plexuses of the gastric antrum and duodenum of normal rats (n = 6) when compared to that of diabetic rats (n = 6). Moreover, the number of VIP-positive neurons was significantly lower in the gastrointestinal tract of diabetic rats compared to normal. The VIP content of the gastric antrum and duodenum of diabetic rat was significantly lower (p< 0.05) than that of normal rat. In contrast to the lower tissue levels of VIP in the gastroduodenal segment of diabetic rats, the plasma level of VIP was significantly higher (p< 0.04) in diabetic rat compared to normal. The plasma level of VIP in normal rats was comparable to that measured in normal human beings. A low tissue level of VIP in the gastroduodenal tract of diabetic rat may contribute in part to the abnormal gut motility observed in diabetic patients. PMID- 11880930 TI - Involvement of cellular calcium in exocrine pancreatic insufficiency during streptozotocin-induced diabetes mellitus. AB - This study investigates the effects of the islet hormones insulin (Ins), glucagon (Glu), and somatostatin (Som) with nerve stimulation (EFS) acetylcholine (ACh) and cholecytokinin-octapeptide (CCK-8) on amylase secretion and intracellular free calcium concentration [Ca(2+)](i) in the pancreas of age-matched control and diabetic rats. Either Ins, Glu or Som elicited small increases in amylase secretion from the pancreas of age-matched control animals compared to a much larger increase in amylase secretion with either EFS, ACh or CCK-8. Combining the islet hormones with either EFS, ACh or CCK-8 resulted in marked potentiation of amylase output. In the diabetic pancreas, the islet hormones had no effect on amylase secretion compared to diabetic control. Moreover, either EFS, ACh or CCK 8 evoked a much smaller increase in amylase output compared to age-matched control. In addition, the islet hormones failed to potentiate the secretory effects of either EFS, ACh or CCK-8. In fura-2 loaded acinar cells from age matched control pancreas either Ins or Glu elicited a small increase in [Ca(2+)](i) whereas Som had no effect. Both ACh and CCK-8 evoked large increases in [Ca(2+)](i) compared to control. Combining either Ins, Glu or Som with either ACh or CCK-8 resulted in a marked elevation in [Ca(2+)](i) compared to the responses obtained with either the islet hormones, ACh or CCK-8 alone. In diabetic fura-2 loaded pancreatic acinar cells, the islet hormones had no effect on [Ca(2+)](i) compared to control and moreover, the responses were much smaller than those obtained in acinar cells from age-matched control. Both ACh and CCK-8 induced large increases in [Ca(2+)]( i) in diabetic acinar cells. However, combining the islet hormones with either ACh or CCK-8 failed to enhance [Ca(2+)](i) compared to the reponses obtained in acinar cells from age-matched control. The results suggests that [Ca(2+)](i) homeostasis is deranged during diabetes mellitus and this in turn is probably associated with reduced pancreatic amylase secretion. PMID- 11880931 TI - Intrauterine growth retardation in experimental diabetes: possible role of the placenta. AB - Fetal growth disorders are common in pregnancy complicated by diabetes. Whereas macrosomia often occurs in infants of diabetic women, growth retardation is almost a rule in spontaneous and experimental diabetes in animals. However, it is not clear when during development growth inhibition starts and how placental pathology might affect fetal growth in maternal diabetes. In this study pregnant Wistar rats were injected (ip) with a single dose of 50 mg/kg of streptozotocin (STZ) on gestation day (GD) 2 and a blood glucose level of 200 mg/dl or more determined 24 hrs later indicated diabetes. The controls were non-treated, buffer treated or, following confirmation of diabetes, injected with a single dose of 2- 6 IU of insulin (Novo Ultralente) once daily. Fetuses and placentae were collected from GD 14--20. Intrauterine growth retardation (IUGR) in STZ group was significant as early as GD 15 and persisted to GD 20. Insulin produced a significant recovery in fetal weight gain. The placentas of STZ-treated group were significantly heavier than those of the control groups. The reduction in cord length of the STZ group became apparent on GD 16 and remained so to term. The placenta of GD 14 STZ group had a thicker decidua basalis and dilated maternal sinusoids. By GD 16, the decidua basalis contained glycogen-containing decidual cells and scattered glycogen cells confirmed by Best's carmine with or without diastase. The glycogen cells of the basal zone were more abundant, and had degenerated in some sites leaving behind cysts with eosinophilic mass. The giant cells had proliferated enormously. The labyrinthine zone appeared spongy with persistent fetal mesenchyme, peri-vascular fibrosis, and enhanced placental barrier. The trophoblasts of the labyrinths also contained traces of glycogen unlike the controls. By GD 18, the decidua basalis of the STZ group was thinner than that of the controls and contained necrotic giant cells and lymphocytic aggregations. In the basal zone, the giant cells had proliferated further; more glycogen cells had degenerated. Perivascular fibrosis was still extensive in the labyrinthine zone. Bloodless maternal sinusoids, extensive vacuolization, degeneration of glycogen islands and formation of cysts characterized the labyrinthine zone. These changes varied in intensity from one area to another in the same placenta and between placentas of the same and of different litters. The development of the upper and lower jaws, elevation and fusion of palatal shelves, reduction of physiological umbilical hernia, descent of the testes, fusion of the urethral folds and separation of digits of the paws were significantly delayed in the STZ group. The consistent association of placental pathology with fetal growth retardation is suggestive of an alteration in placental function possibly contributing to IUGR in STZ-induced diabetes in rats. PMID- 11880932 TI - A clinical trial of chronic care diabetic clinics in general practice in the United Arab Emirates: a preliminary analysis. AB - Control of diabetes mellitus is a high priority for primary health care systems. One innovative method of diabetes care delivery is the use of structured diabetes care in primary care. This includes the use of chronic care diabetes clinics or mini-clinics operated by general practitioners in primary care. There is limited experience with this model in non-Western settings. This study sought to evaluate a multi-component structured approach to diabetes care in primary care including chronic care diabetes clinics in a newly developed country in the Arabian Gulf. The study design used was a controlled before-after methodology. Three primary health centers were chosen for the intervention with six of the remaining clinics in a Health District being used as controls. A multifaceted intervention was initiated in the intervention clinics composed of chronic care diabetes clinics, a diabetic flow chart, and educational programs for clinic nurses and doctors and patients. The study intervention took place over a period of 18 months with three diabetic outcomes (fasting blood glucose, blood pressure and cholesterol) and adherence to seven diabetes guidelines being compared for the year prior to the intervention and during the last 12 months of the intervention period. Knowledge and satisfaction questionnaires were also administered to intervention and control subjects at the end of the study. In this study, 219 subjects were enrolled (130 males and 89 females). They had a mean age of 51.6 years and a mean of 3.1 years of formal education. Of these 109 were enrolled in one of three clinics that had a chronic care diabetes clinic and 110 were enrolled in one of the six control clinics. Subjects had diabetes for a mean of 7.8 +/- 4.8 years and the majority was treated with pharmacological therapy. Baseline characteristics in the intervention and the control clinics were similar with the exception of younger age (p = 0.01) and a trend for more males (p = 0.06) in the intervention clinics. There was a statistically insignificant change noted with the intervention in the three clinical outcomes studied (fasting blood glucose, blood pressure and cholesterol) both in comparison to the control group before and after and within the intervention group. However most changes noted were in the expected direction of improvement; six of the seven guidelines were statistically improved in the intervention group when compared with the control group. Within the intervention group, adherence with five of seven guidelines was also statistically significantly increased with the remaining guidelines showing a trend in favor of improvement (fasting blood glucose measurements (p = 0.07) and urine determinations for protein (p = 0.07)). Knowledge questionnaire scores were similar between the intervention and control groups on completion of the study but 2 of 4 items on a satisfaction scale were statistically significantly higher in the intervention group. The intervention described in this setting was successful in improving adherence to diabetes guidelines and increased some aspects of satisfaction with diabetes care. The intervention did not result in a statistically significant improvement in clinical outcomes but changes noted were in the expected direction of improvement. The significant improvement in adherence to diabetes guidelines suggests that this intervention is a promising model for diabetes care for newly developed countries. PMID- 11880933 TI - Diabetes mellitus and its complications in a Hungarian population. AB - The aim of this study was to examine the disease characteristics and complications of diabetes mellitus in patients in a Hungarian rural community. Data relating to age, sex, date of onset of diabetes, fasting blood glucose values and all diseases associated with diabetics were retrieved from the medical records of patients. Almost six percent (5.7%) of the population has diabetes mellitus. The percentage of Type I diabetic patients in this population was 5.8 percent. The prevalence of diabetes was slightly but not significantly higher in females than in males. The mean age of the diabetic population was 52.1 +/- 11.3 for male and 53.47 +/- 15.7 for the female patients. The peak age of onset of diabetes mellitus was in the sixth decade of life. The mean fasting blood sugar value was 10.64 +/- 0.6 and 10.57 +/- 0.5 mmol L(-1), in male and female diabetic patients (n = 103), respectively. Diabetic patients presented with many signs and symptoms in the general practice setting. The findings of this study showed that diabetics present with many disease conditions and signs and symptoms in the general practice setting. Many of these conditions are known to be associated with diabetes while others are not. As a result of the adverse effects of diabetes mellitus on the cardiovascular system and on body metabolism as a whole, the damage and morbidity caused by diabetes mellitus may have been underestimated. The results of this study have shed light on the unrecognised complications of diabetes mellitus. PMID- 11880934 TI - The origin of the azygos venous system, as deduced from an anatomical and radiological study employing a corrosion technique. AB - The aim of this study is not only to describe the origin of the human azygos venous system by performing a 3-D reconstruction of a CT scan but also to evaluate the value of the techniques employed in investigating the topographical anatomy of a venous system in the body. Following perfusion with saline to wash away the blood, we injected an ALTUFIX/MINIUM mixture into the azygos vein of a cadaver. The head and trunk were subsequently corroded with hydrochloric acid (HCl). A CT scan of the trunk was obtained both before and after corrosion. According to the spatial resolution of the CT scan, the thinnest identifiable detail was measurable as 0.5 mm. The vertebral lumbar venous system was described, specifying the nomenclature of the lumbar veins (the lumbar veins being designated according to the vertebral body along which they run). On the right side, the lumbar veins at L2 and at L3 formed the lateral root of the azygos vein. On the left side, the vein at L2 formed the reno-azygo-lumbar arch (of Lejars). The lumbar veins, and the origin of the azygos system, were described and compared with previous studies. The 3-D reconstruction showed the importance of veins associated with the posterior paravertebral muscles. This description poses the problem of the metamerisation of the veins, but further evidence is required. Comparisons of the CT scans, 3-D reconstructions, and the ALTUFIX models of the veins obtained from the corrosion technique allowed verification of the 3-D reconstruction and correction of the errors inherent in a computer reconstruction. It is concluded that the description, and understanding, of such a complex system as the vertebral venous system is more valid when the results obtained using different techniques are compared. PMID- 11880935 TI - Neuroendocrine epithelial cells in the broncho-parabronchial transition of embryonic, immature and mature quail (Coturnix coturnix). AB - The main objective of this integrated light microscopic, transmission and scanning electron microscopic study was to describe in greater detail the structural pattern and developmental stages of pulmonary neuroendocrine epithelial cells (PNECs) in the broncho-parabronchial transition (BPT) of both developing and mature quail. In mature quail the BPT appeared as a diaphragm opening into the parabronchial vestibulum. Perpendicular sections revealed two bilayered crest-like entrance folds invested by a uniform population of granular cells with lamellar bodies and a brush border of blunt microvilli. Solitary PNECs were found interlaced between the granular cells on both sides of the BPT. In addition, PNECs with small dense-cored vesicles (DCVs) were found subepithelially in the lamina propria mucosae of the parabronchial compartment of the BPT, which was surrounded by a capsule of granular cells. Furthermore, clusters of PNECs devoid of any epithelial capsule, but with large DCVs were located in the tunica propria mucosae. The first signs of the developing BPT could be identified as early as embryonic day (ED) 13. By ED 17, the area of the future BPT was seen to be invested by a uniform population of granular cells but the entrance fold proper did not appear until after hatching. Solitary or clustered PNECs were demonstrated in parabronchial buds growing into the mesenchyme on ED 12 and 13: either type I PNECs with small DCVs measuring about 80-120 nm or type II PNECs with large dense granules measuring approximately 150-280 nm. Both types of PNECs represented a temporary cell population in the prehatching period. In the parabronchus no PNECs or neuroendocrine epithelial bodies could be demonstrated in mature and immature quail beyond the BPT. PMID- 11880936 TI - Atrial natriuretic peptide (ANP) system in frog skin. AB - In this study the ultrastructure of Rana esculenta skin is described. Cytochemical methods were used to localize guanylate cyclase in the presence of atrial natriuretic peptide and immunocytochemical methods showed the presence of the atrial natriuretic peptide in various levels of skin. The peptide is mainly found in the epithelium and in the lymph sacs of the tela subcutanea. Its receptors are located in the same zones and are indicated by guanylate cyclase activity. We demonstrate that frog skin is a target organ for atrial natriuretic peptide and propose that, at this level, the peptide carries out an important osmoregulatory role. PMID- 11880938 TI - Introduction to the symposium: teaching of anatomy/embryology. PMID- 11880940 TI - Curricular change--cutting anatomy to the core. PMID- 11880939 TI - Blueprint 1994: follow-up and implications. PMID- 11880941 TI - Teaching anatomy; a clinicians view. AB - The General Plan Anatomy as defined by the Netherlands Association of Anatomists and published in the European Journal of Morphology in 1999 is a good starting point for discussion between anatomists and clinicians to define what is indispensable anatomical knowledge for a competent physician. The plan to teach anatomy within a clinical context and to relate form and structure on function makes learning anatomy more attractive and will have a greater appeal for the student. The list of discipline-related objectives is too extensive for a general practitioner whereas parts of this list are too limited for related medical specialists. Teaching anatomy and teaching clinical sciences and expertise must become more integrated. PMID- 11880942 TI - Evaluation of the medical curriculum: why, when, by whom and for whom should questionnaires be used. AB - The undergraduate medical curriculum has been modified or even totally reorganized in many countries in recent years, and there are plans to make departmental budgets and the salaries of university professors partially dependent on the outcome of teaching. Questionnaires are often used in such situations as a means of curriculum evaluation. Based on our own experience such evaluations should be done not only during and immediately after a course in the curriculum, but also at later time points, e.g., at the end of the undergraduate and also the postgraduate phase. The clinical relevance of lectures and courses can only be graded adequately after some years of clinical experience. Gross anatomy was graded top at all time points evaluated and reached higher levels of 'clinical relevance' than other typical preclinical and even clinical subjects. Efforts should be made to obtain a high response rate for representative results. After modifying parts of a course detailed questionnaires should also include space for students' suggestions. The results of such evaluations are not only relevant to the head of department as feedback on the individual lecturers but also important for the curriculum committee and the dean. Anatomists should utilize these evaluations to improve teaching. PMID- 11880945 TI - [Neuropsychological assessment of malingering]. AB - Current problems and neuropsychological assessment strategies of malingering detection (assessment of negative response bias or non-optimal test behaviour) are reviewed. First, the paper discusses major conceptual problems inherent in the definition of malingering, factitious disorders, and somatoform disorders. Traditional and modern test approaches and diagnostic procedures are reviewed. Two case vignettes illustrate the application of particular strategies for malingering assessment. In German speaking countries, malingering research has not yet aroused profound interest comparable to that in English speaking countries. Diagnostic standards and instruments still have to be refined. PMID- 11880944 TI - [The concept of hyperthymia]. AB - The article reviews the conceptual history of "hyperthymia". Since K. W. Stark had used this term in the early 19(th) century, it has developed in two different directions: (1) to delineate a psychopathological syndrome and (2) to define a type of personality disorder (psychopathy). As Kurt Schneider's personality disorder (psychopathy) concept was easily understood and highly practicable, it became influential during the 20(th) century. Earlier before, psychiatrists such as E. Mendel, C. Wernicke and C. G. Jung had described entities such as "chronic mania", "hypomania" or "sanguinic degeneration", which were rather similar to each other. We analyze the historical development of such concepts. Emil Kraepelin was highly influential, as he introduced "constitutional excitation" into a broad concept of manic-depressive illness and saw it as a very mild form. After Kraepelin such spectrum concept was first forgotten. Only in recent years these historical considerations were confirmed by empirical observations, although a separate hyperthymic disorder is neither part of DSM-IV nor ICD-10. The concept of a hyperthymic temperament or a hyperthymic personality is a trait marker and should be differentiated from hypomania as a state-marker. Nowadays, the importance of hyperthymia is not so much one of a disorder requiring treatment; rather the concept has interesting genetic, diagnostic and conceptual consequences. PMID- 11880946 TI - [Atypical neuroleptics in the treatment of aggression and hostility in schizophrenic patients]. AB - The treatment of aggressive symptoms in schizophrenic patients is a relevant clinical problem. We systematically review the efficacy of the different atypical neuroleptics on acute or persistent aggressive symptoms also regarding methodological problems. Currently typical neuroleptics are still first choice in treating acute aggressive symptoms, while risperidone and olanzapine could be alternatives. In persistent aggression clozapin shows the best specific results. Typical depot neuroleptics should be considered in cases when medication compliance is a problem. PMID- 11880947 TI - [Vascular dementias]. AB - Cerebrovascular disorders are the second most frequent cause of dementia in late life next to Alzheimer's disease. A recent community-based autopsy study has demonstrated that relevant cerebrovascular changes are much more prevalent in individuals aged 70+ years than previously assumed. Furthermore, the combination between cerebrovascular lesions and Alzheimer-type pathology is the most common neuropathological finding in elderly patients with dementia. There is still some uncertainty about which types of cerebrovascular changes are most likely to cause cognitive impairment including dementia and which pathogenetic mechanisms are involved. Without doubt, however, the vascular dementias are a heterogeneous group of diseases in terms of etiology, histopathology, and clinical appearance. According to the vessel calibres and perfusion territories that are preferentially affected a distinction is commonly made between the frequent subcortical small-vessel disease and the rare cortical large-vessel disease. With these morphological subtypes three major clinical variants are associated: dementia due to subcortical lacunes and white matter changes including Binswanger's disease, multi-infarct-dementia, and dementia due to singular strategic infarcts. In most cases of dementia of cerebrovascular origin the pattern of intellectual impairment is frontal or subcortical, in contrast to the typical cortical presentation of Alzheimer's disease. Deterioration of executive function and attention as well as changes in personality, rather than memory loss, are the predominant symptoms. Therefore the current diagnostic criteria for dementia are poorly suited for the detection of vascular dementias. None of the criteria that have been specifically proposed for the diagnosis of vascular dementias provide clear guidelines for evaluating the causal relationship between cerebrovascular lesions and psychopathological findings. Further research will reveal whether clinical diagnosis can be improved by taking into account the heterogeneity of cerebrovascular diseases. A large proportion of dementias of cerebrovascular origin may be preventable by treating the risk-factors for stroke. Once significant cognitive impairment has occurred, however, there is no established pharmacological treatment for the vascular dementias to date. Only recently results of placebo-controlled clinical trials have become available showing that cholinergic treatment strategies are effective in vascular dementia and in dementia due to combined vascular and neurodegenerative pathologies. PMID- 11880948 TI - [Interpersonal psychotherapy (IPT) of depression: an overview of the psychiatric historical background and clinical practice of IPT]. AB - Among the forms of short-term psychotherapies for the treatment of depression, IPT (16 - 20 sessions) by Klerman u. Mitarb. is meanwhile one of the most well known and clinically practiced approaches. IPT has also been carefully controlled in a variety of studies proving efficacy. Interpersonal psychotherapy is based on the ideas of the interpersonal school of psychiatry. The hypothesis is that psychiatric illnesses and here depression develop in an interpersonal context. Life problems may contribute to onset and potentially chronicity of (current) depression and vice versa. By enhancing interpersonal functioning of the depressed patient IPT initiates the reduction of depressive symptomatology and helps solving of current life problems as dual goal of therapy. PMID- 11880949 TI - Ascertainment-adjusted parameter estimates revisited. AB - Ascertainment-adjusted parameter estimates from a genetic analysis are typically assumed to reflect the parameter values in the original population from which the ascertained data were collected. Burton et al. (2000) recently showed that, given unmodeled parameter heterogeneity, the standard ascertainment adjustment leads to biased parameter estimates of the population-based values. This finding has important implications in complex genetic studies, because of the potential existence of unmodeled genetic parameter heterogeneity. The authors further stated the important point that, given unmodeled heterogeneity, the ascertainment adjusted parameter estimates reflect the true parameter values in the ascertained subpopulation. They illustrated these statements with two examples. By revisiting these examples, we demonstrate that if the ascertainment scheme and the nature of the data can be correctly modeled, then an ascertainment-adjusted analysis returns population-based parameter estimates. We further demonstrate that if the ascertainment scheme and data cannot be modeled properly, then the resulting ascertainment-adjusted analysis produces parameter estimates that generally do not reflect the true values in either the original population or the ascertained subpopulation. PMID- 11880950 TI - Quantitative-trait homozygosity and association mapping and empirical genomewide significance in large, complex pedigrees: fasting serum-insulin level in the Hutterites. AB - We present methods for linkage and association mapping of quantitative traits for a founder population with a large, known genealogy. We detect linkage to quantitative-trait loci (QTLs) through a multipoint homozygosity-mapping method. We propose two association methods, one of which is single point and uses a general two-allele model and the other of which is multipoint and uses homozygosity by descent for a particular allele. In all three methods, we make extensive use of the pedigree and genotype information, while keeping the computations simple and efficient. To assess significance, we have developed a permutation-based test that takes into account the covariance structure due to relatedness of individuals and can be used to determine empirical genomewide and locus-specific P values. In the case of multivariate-normally distributed trait data, the permutation-based test is asymptotically exact. The test is broadly applicable to a variety of mapping methods that fall within the class of linear statistical models (e.g., variance-component methods), under the assumption of random ascertainment with respect to the phenotype. For obtaining genomewide P values, our proposed method is appropriate when positions of markers are independent of the observed linkage signal, under the null hypothesis. We apply our methods to a genome screen for fasting insulin level in the Hutterites. We detect significant genomewide linkage on chromosome 19 and suggestive evidence of QTLs on chromosomes 1 and 16. PMID- 11880951 TI - Distinct BRCA1 rearrangements involving the BRCA1 pseudogene suggest the existence of a recombination hot spot. AB - The 5' end of the breast and ovarian cancer-susceptibility gene BRCA1 has previously been shown to lie within a duplicated region of chromosome band 17q21. The duplicated region contains BRCA1 exons 1A, 1B, and 2 and their surrounding introns; as a result, a BRCA1 pseudogene (PsiBRCA1) lies upstream of BRCA1. However, the sequence of this segment remained essentially unknown. We needed this information to investigate at the nucleotide level the germline deletions comprising BRCA1 exons 1A, 1B, and 2, which we had previously identified in two families with breast and ovarian cancer. We have analyzed the recently deposited nucleotide sequence of the 1.0-Mb region upstream of BRCA1. We found that 14 blocks of homology between the tandemly repeated copies (cumulative length = 11.5 kb) show similarity of 77%-92%. Gaps between blocks result from insertion or deletion, usually of repetitive elements. BRCA1 exon 1A and PsiBRCA1 exon 1A are 44.5 kb apart. In the two families with breast and ovarian cancer mentioned above, distinct homologous recombination events occurred between intron 2 of BRCA1 and intron 2 of PsiBRCA1, leading to 37-kb deletions. Breakpoint junctions were found to be located at close but distinct sites within segments that are 98% identical. The mutant alleles lack the BRCA1 promoter and harbor a chimeric gene consisting of PsiBRCA1 exons 1A, 1B, and 2, which lacks the initiation codon, fused to BRCA1 exons 3-24. Thus, we report a new mutational mechanism for the BRCA1 gene. The presence of a large region homologous to BRCA1 on the same chromosome appears to constitute a hot spot for recombination. PMID- 11880952 TI - Do the benefits of varicella vaccination outweigh the long-term risks? A decision analytic model for policymakers and pediatricians. AB - Although varicella vaccine is recommended for infants, many physicians and parents have withheld vaccination from infants because of concerns about the vaccine's long-term efficacy. We used a decision-analytic Markov model to examine the effects of decreasing vaccine efficacy on individuals and society. The model incorporated published data on age-specific incidence, morbidity, and mortality rates, as well as data on shifting disease burden from childhood to adulthood as vaccine compliance increases. The effects of 2 vaccination strategies-- vaccinating infants at age 12 months and waiting to vaccinate until children are 10 years of age---were compared with the effects of no vaccination. If the efficacy of the vaccine were to decrease by 75%, then 50% compliance with vaccination at age 12 months would save 1800 life-years and 12,800 quality adjusted life-years annually in the United States. The quality-adjusted life expectancy of individuals vaccinated at age 12 months would be 63 h longer than that of nonvaccinated individuals and would increase to 79 h as vaccination compliance increases and the burden of chickenpox shifts to adulthood. Varicella vaccination of infants at age 12 months appears to be beneficial, even if the efficacy of the vaccine declines substantially. PMID- 11880953 TI - Usefulness of procalcitonin as a marker of systemic infection in emergency department patients: a prospective study. AB - We prospectively evaluated serum procalcitonin concentrations in patients who presented to an emergency department (ED) with suspected infectious or inflammatory disease. Of 195 study patients, 68 had final diagnosis of systemic infection, and 24 of those 68 had elevated serum procalcitonin levels (>0.5 ng/mL). The procalcitonin level had a sensitivity of 0.35 and specificity of 0.99 for the diagnosis of systemic infection. In multivariate analysis, the procalcitonin level was the only independent variable associated with this diagnosis; in contrast, the C-reactive protein level was not. All patients with systemic infections who ultimately died had procalcitonin levels of >0.5 ng/mL at admission. Procalcitonin levels were significantly higher in patients who ultimately died of systemic infection than in patients who survived. The optimal procalcitonin threshold for the ED population may be lower than that proposed for critically ill patients. Determination of the procalcitonin level may be useful for screening and prognosis of more-severely ill ED patients. PMID- 11880954 TI - Clinical determinants for the recovery of fungal and mezlocillin-resistant pathogens from bile specimens. AB - We conducted a retrospective analysis of all bile specimens obtained for routine cultures from January 1995 through December 1999 at our tertiary care hospital. Results of microbiologic testing were linked to clinical parameters gathered by means of chart review. A total of 722 isolates were cultured from 345 of 454 bile specimens obtained from 288 individual patients. Prior receipt of a >7-day course of antibiotics (odds ratio [OR], 5.7), extensive leukocytosis (leukocyte count, >20,000 cells/microL) on admission (OR, 7.8), endoscopic or percutaneous biliary manipulation during the previous 14 days (OR, 2.9), and treatment in an internal medicine ward (OR, 2.5) were independent factors significantly associated (Pless than-or-eq, slant.05) with recovery of Candida species from bile specimens. Culture of mezlocillin-resistant bacteria from bile specimens was independently associated with the specimen having been obtained >1 week after admission (OR, 3.8), lack of history of endoscopic biliary drainage (OR, 3.2), and high serum aspartate aminotransferase levels (>72 U/L) on admission (OR, 2.6). Prospective studies are warranted to evaluate accordingly adjusted empiric therapies for biliary infections. PMID- 11880955 TI - Epidemiology and outcome of mould infections in hematopoietic stem cell transplant recipients. AB - Reports have focused on the emergence of moulds as pathogens in recipients of hematopoietic stem cell transplants. To review the incidence of and risks for mould infections, we examined the records of 5589 patients who underwent hematopoietic stem cell transplantation at the Fred Hutchinson Cancer Research Center (Seattle) from 1985 through 1999. After 1992, the incidence of invasive aspergillosis increased in allograft recipients and remained high through the 1990s. Infections with non-fumigatus Aspergillus species, Fusarium species, and Zygomycetes increased during the late 1990s, especially in patients who received multiple transplants. Although infection caused by Scedosporium species was common in patients who had neutropenia, infection caused by Zygomycetes typically occurred later after transplantation, when patients had graft-versus-host disease. The overall 1-year survival rate was equally poor (similar20%) for all patients with mould infections. The results of the present study demonstrate the changing epidemiology of mould infections, emphasizing the increasing importance of amphotericin B--resistant organisms and the differences in risks and outcome of infection with different filamentous fungi. PMID- 11880956 TI - Intestinal microsporidiosis due to Enterocytozoon bieneusi in elderly human immunodeficiency virus--negative patients from Vigo, Spain. AB - We report what is, to our knowledge, the first study in which microsporidial infection was detected in elderly human immunodeficiency virus (HIV)--negative patients. Of the 60 elderly patients studied, 47 had diarrhea. Intestinal microsporidiosis due to Enterocytozoon bieneusi was diagnosed in 8 patients (17.02%) by use of Weber's chromotrope-based stain and polymerase chain reaction with species-specific primers. The mean age of these 8 patients was 75 years; 7 had chronic diarrhea and 1 had nonchronic diarrhea. Six of the patients with chronic diarrhea had no other pathogens isolated. In our opinion, elderly patients, because of their special immunological characteristics, should be considered a group at risk for the acquisition of intestinal microsporidiosis. PMID- 11880957 TI - Clinical outcomes for patients with bacteremia caused by vancomycin-resistant enterococcus in a level 1 trauma center. AB - To assess the degree of morbidity and mortality attributable to vancomycin resistance in enterococcal bacteremia (EB), a matched case-control study was conducted. Patients with bacteremia due to vancomycin-resistant enterococcus (VRE) were matched to control patients with bacteremia due to vancomycin susceptible enterococcus. During 1996--2000, 65 patients with cases of clinically significant VRE bacteremia were identified, and 53 of these patients were successfully matched. In this group of patients, VRE bacteremia was found to be an independent predictor of crude mortality (odds ratio [OR], 4.0; 95% confidence interval [CI], 1.2--13.3) and the infection-related mortality rate (OR, 5.2; 95% CI, 1.4--20.0). It was also an independent predictor of the rate of clinical failure at day 7 after the onset of EB (OR, 4.6; 95% CI, 1.2--17.3) and overall clinical failure (OR, 4.3; 95% CI, 1.3--14.5) and was associated with a longer mean length of hospitalization after the onset of EB, compared with that for control patients (22.7plus minus1.88 vs. 15.9 +/- 1.7, P=.006). These observations indicate that vancomycin resistance in EB independently affects outcomes. PMID- 11880958 TI - Delayed reimplantation arthroplasty for candidal prosthetic joint infection: a report of 4 cases and review of the literature. AB - Fungal prosthetic joint infection (PJI) is rare, with Candida species being the most frequently reported pathogen in the medical literature. The risk of relapse following delayed reimplantation arthroplasty for candidal PJI is unknown. We describe 4 new cases and summarize 6 previously reported cases of candidal PJI treated with delayed reimplantation arthroplasty. Ninety percent of the patients received antifungal therapy. Eight patients received amphotericin B either alone or in combination with other antifungals. One patient received fluconazole alone. The median duration of time from resection arthroplasty to reimplantation for total hip and total knee arthroplasties was 8.6 and 2.3 months, respectively. Eight patients did not have relapse of candidal PJI following delayed reimplantation arthroplasty after a median duration of follow-up of 50.7 months (range, 2--73 months). Candidal PJI can be successfully treated with delayed reimplantation arthroplasty after receipt appropriate antifungal therapy. PMID- 11880959 TI - Invasive aspergillosis in allogeneic stem cell transplant recipients: increasing antigenemia is associated with progressive disease. AB - The kinetics of serum Aspergillus galactomannan, as determined by enzyme-linked immunosorbent assay, was examined in 37 allogeneic stem cell transplant (SCT) recipients treated for invasive aspergillosis (IA). Fifty-eight periods of response ("response episodes") were evaluated. There were 42 response episodes that were considered "treatment failures" and 16 that were considered "good" (that is, complete or partial) responses. At baseline (the first day of each new response episode), the patients who experienced treatment failure and those who had good responses did not differ significantly with regard to median galactomannan index (GMI) value. Thereafter, GMI values significantly increased in the treatment failure group, whereas no significant changes were observed in the good response group (P=.002). An increase in the GMI value of 1.0 over the baseline value during the first week of observation was predictive of treatment failure with a sensitivity of 44%, a specificity of 87%, and a positive predictive value of 94%. We conclude that serial determination of serum GMI values is a useful tool for assessing prognosis of IA in allogeneic SCT recipients during treatment. PMID- 11880960 TI - Two-day regimen of acyclovir for treatment of recurrent genital herpes simplex virus type 2 infection. AB - The standard course of antiviral therapy for recurrent genital herpes requires administration of multiple doses of medication for 5 days. To assess the efficacy of a shorter course of antiviral therapy, patients with recurrent genital herpes simplex virus type 2 (HSV-2) infection were enrolled in a randomized, double blind, placebo-controlled study of acyclovir (800 mg given by mouth 3 times per day [t.i.d.]) for 2 days. Of 131 people enrolled in the study, 84 (51 women and 33 men) were observed for >/=1 recurrence and 65 were observed for 2 recurrences, for which the patient was administered the same study drug (acyclovir or placebo). Acyclovir therapy (800 mg given by mouth t.i.d. for 2 days) significantly reduced the duration of lesions (median for acyclovir versus placebo, 4 days versus 6 days; P=.001), episode (4 days versus 6 days; P<.001), and viral shedding (25 hours versus 58.5 hours; P=.04), and it increased the proportion of aborted episodes (P=.029). A 2-day course of acyclovir is a convenient alternative for treatment of recurrent genital herpes. PMID- 11880961 TI - Are Haemophilus influenzae infections a significant problem in India? A prospective study and review. AB - It has been suggested Haemophilus influenzae serotype b (Hib) disease is uncommon in Asia. During 1993--1997, we conducted prospective surveillance of acute infections caused by H. influenzae in 6 academic referral Indian hospitals. The study included 5798 patients aged 1 month to 50 years who had diseases likely to be caused by H. influenzae; 75% of the patients were aged <5 years. A total of 125 H. influenzae infections were detected, 97% of which were caused by Hib. Of 125 isolates, 108 (86%) were from children aged <5 years, and 11 (9%) were from adults aged >18 years. Sixty-two percent of the patients had meningitis. The case fatality rate was 11% overall and 20% in infants with Hib meningitis. Up to 60% of all isolates were resistant to chloramphenicol, ampicillin, trimethoprim sulfamethoxazole, or erythromycin; 32% were resistant to >/= 3 antimicrobial drugs, but none were resistant to third-generation cephalosporins. These data suggest that available Hib vaccines will benefit Indian children. PMID- 11880963 TI - Infectious disease experimentation involving human volunteers. AB - The current care of patients with infectious diseases owes a tremendous debt to healthy volunteers who allowed investigators to induce disease in them for the study of transmission, natural history, and treatment. We reviewed the English language medical literature about the rarely discussed subject of the use of healthy volunteers in human-subject research in infectious diseases to determine the contributions of these experiments to the current understanding of disease transmission. The literature review focused on hepatitis, upper respiratory infections, and malaria, which represent the array of issues involved in this type of research. Researchers successfully induced infection through injecting, nebulizing, and feeding specimens to thousands of volunteers, who included authentic volunteers as well as soldiers and imprisoned subjects. These volunteers often undertook unforeseen and unpredictable risks during these experiments for the benefit of others. Future research in these areas must strike an adequate balance between the risks to participants and the benefits to society. PMID- 11880962 TI - Valacyclovir for episodic treatment of genital herpes: a shorter 3-day treatment course compared with 5-day treatment. AB - Valacyclovir given in a 5-day regimen of 500 mg twice per day is effective as short-term treatment of episodes of recurrent genital herpes. This study compared the efficacy of a shorter, 3-day course (for 402 patients) with that of a 5-day course (for 398 patients) of valacyclovir for persons with frequent recurrence of symptoms. No significant differences were detected between the 2 dosing schedules for any of the end points measured. Median times to lesion healing, of pain duration, and of episode length for the 5-day versus 3-day treatment were 4.7 versus 4.4 days, 2.5 days versus 2.9 days, and 4.4 days versus 4.3 days, respectively. The proportions of patients with aborted lesions were 26.6% and 25.4% in the 5-day and 3-day groups, respectively. A 3-day course of 500 mg of valacyclovir administered twice daily as episodic treatment of recurrent genital herpes is equivalent to a 5-day course with regard to key markers of efficacy. PMID- 11880964 TI - Shining light on "Dark Winter". AB - On 22--23 June 2001, the Johns Hopkins Center for Civilian Biodefense Strategies, in collaboration with the Center for Strategic and International Studies, the Analytic Services Institute for Homeland Security, and the Oklahoma National Memorial Institute for the Prevention of Terrorism, held a senior-level exercise entitled "Dark Winter" that simulated a covert smallpox attack on the United States. The first such exercise of its kind, Dark Winter was constructed to examine the challenges that senior-level policy makers would face if confronted with a bioterrorist attack that initiated outbreaks of highly contagious disease. The exercise was intended to increase awareness of the scope and character of the threat posed by biological weapons among senior national security experts and to bring about actions that would improve prevention and response strategies. PMID- 11880965 TI - Directly observed therapy for the treatment of people with human immunodeficiency virus infection: a work in progress. AB - The principle of directly observed therapy (DOT) has its roots in the treatment of tuberculosis (TB), for which DOT programs have improved cure rates in hard-to reach populations. Human immunodeficiency virus (HIV) and TB affect similar populations, and there are concerns about both regarding the development of drug resistance associated with poor adherence to therapy. Accordingly, DOT may benefit certain HIV-infected people who have difficulty adhering to highly active antiretroviral therapy. However, important differences exist in the treatment of these diseases that raise questions about how DOT can be adapted to HIV therapy. DOT for management of HIV infection has been effective among prisoners and in pilot programs in Haiti, Rhode Island, and Florida. Although DOT can successfully treat HIV infection in marginalized populations in the short term, a multitude of questions remain. This review provides an account of the preliminary development of DOT programs for the treatment of HIV-infected individuals. PMID- 11880966 TI - Nucleoside-analogue reverse-transcriptase inhibitors plus nevirapine, nelfinavir, or ritonavir for pretreated children infected with human immunodeficiency virus type 1. AB - The relative potency and tolerability of multidrug regimens used to treat infants and children infected with human immunodeficiency virus type 1 (HIV-1) are largely unknown. In Pediatric AIDS Clinical Trials Group (PACTG) Protocol 377, 181 infants and children were assigned to receive stavudine (d4T) plus nevirapine (NVP) and ritonavir (RTV); d4T plus lamivudine (3TC) and nelfinavir (NFV); d4T plus NVP and NFV; or d4T plus 3TC, NVP, and NFV. Eleven additional children received d4T and NVP plus NFV given twice daily. All subjects had not previously received protease inhibitors or nonnucleoside reverse-transcriptase inhibitors and all had been immunologically stable while receiving reverse-transcriptase inhibitor therapy. After 48 weeks of therapy, 17 (41%) of 41 subjects receiving d4T-NVP-RTV, 13 (30%) of 44 receiving d4T-NVP-NFV, 21 (42%) of 50 receiving d4T 3TC and NFV (3 times daily), and 22 (52%) of 42 receiving d4T-3TC-NVP-NFV were still receiving their assigned therapy and had HIV-1 RNA suppression to /= 96 weeks in a prospective, open, uncontrolled multicenter trial. The pharmacokinetics of indinavir and of nelfinavir were determined and showed large interindividual differences. After 96 weeks of therapy, 69% and 50% of the patients had an HIV-1 RNA load that was below the HIV assays' detection limits of 500 and 40 copies/mL, respectively. Virologic failure was associated with poor compliance and younger age (independent of baseline virus load and receipt of pretreatment). Relative CD4 cell counts increased significantly in relation to the median of the age-specific reference value, from a median of 44% at baseline to 94% after 96 weeks. In a high percentage of the children, clinical, virologic, and immunologic response rates to combination therapy were optimal during the initial 2 years of therapy. PMID- 11880969 TI - Remission of cutaneous Mycobacterium haemophilum infection as a result of antiretroviral therapy in a Human Immunodeficiency Virus--infected patient. AB - We describe the first Mycobacterium haemophilum infection that occurred in a patient with human immunodeficiency virus in Germany and report 7 newly diagnosed cases of M. haemophilum infection. In the former case, a local M. haemophilum skin infection resolved as a result of successful antiretroviral therapy only; however, that clinical outcome may not be possible for more invasive forms of the disease. PMID- 11880970 TI - Left ventricular dysfunction in children with fulminant enterovirus 71 infection: an evaluation of the clinical course. AB - We describe 2 children with typical hand, foot, and mouth disease due to enterovirus 71 infection, 1 of whom died. Both cases were complicated by acute fulminant shock syndrome; the patients had remarkable acute left ventricular dysfunction. The clinical experience indicates that the rapid death associated with fulminant enterovirus rhombencephalitis is the result of rapid cardiogenic shock rather than neurogenic pulmonary edema. PMID- 11880971 TI - Assessment of the Binax NOW Streptococcus pneumoniae urinary antigen test in children with nasopharyngeal pneumococcal carriage. AB - We evaluated the Binax NOW Streptococcus pneumoniae urinary antigen assay by testing 210 healthy children aged 2--60 months living in urban slums of Quito, Ecuador. Healthy children with nasopharyngeal carriage of S. pneumoniae were significantly more likely to have positive urinary antigen test results than were children who were not carriers (30 of 138 vs. 3 of 71 children; chi2=10.8; P<.001). The rate of nasopharyngeal carriage of S. pneumoniae decreased with increasing age; the lowest rates were found in children with the worst nutritional status. PMID- 11880973 TI - Milk intake and energy expenditure of free-ranging northern fur seal, Callorhinus ursinus, pups. AB - Milk ingested by mammalian offspring, coupled with offspring's utilization of this energetic investment, influences survival and growth. A number of studies have examined milk intake in otariids, but few have examined milk intake over the entire lactation period, and none has independently measured energy expenditure concurrent with milk intake. We concurrently examined milk intake, field metabolic rate (FMR), and body composition of 41 pups over the entire lactation interval in 1995 and 1996 on St. Paul Island, Alaska. One hundred two metabolic measurements were obtained with isotope dilution methods. Mean milk intake did not differ annually but increased with age and mass, ranging from 3,400+/-239 to 6,780+/-449 (+/-SE) mL per suckling bout. Milk energy consumption did not vary with age on a mass-specific basis. No differences were detected in milk volume consumed by male and female pups, either absolutely or on a mass-specific basis. Mass-specific FMR peaked during molting, was lowest postmolt, and did not vary by sex. Pups in 1995 had lower FMR than pups in 1996 and were also fatter. Mean milk energy utilized for maintenance metabolism decreased over time from 77% to 43% in 1995 and remained at 71% in 1996. Pup body mass was negatively correlated with the percentage of total body water and positively correlated with the percentage of total body lipid (TBL). Pups increased the percentage of TBL from 16% to 37%. Northern fur seal pups increased energy intake over lactation, while concurrent changes in body composition and pelage condition resulted in mass-specific metabolic savings after the molt. PMID- 11880974 TI - The effect of dietary fat content on lactation energetics in the European hare (Lepus europaeus). AB - European hares selectively feed on plants with high fat and hence energy content. We hypothesized that these dietary requirements limit the ability of hares to adjust daily food intake during periods of high energy requirements, namely lactation. Our measurements in captive lactating females show that does kept on a low-fat diet increased food intake compared to does on a high-fat diet but assimilated significantly lower amounts of energy. Further, does fed a low-fat diet showed a prolonged rise of food intake during lactation, reduced milk energy content and lower milk mass production at large litter sizes. We hypothesize that impaired milk production under suboptimal fat supply is due to the inability of females to increase the capacity of nutrient-processing organs rapidly enough to meet the high energy demands of precocial juveniles with high metabolic costs. Thus, in hares, the production of precocial young may be viewed as a constraint, caused by their inability to dig thermally buffered burrows, rather than as an adaptive reproductive strategy. We suggest that the interaction of lactation energetics, dietary requirements, and reduced plant diversity in modern agricultural landscapes has facilitated the decline of hare populations across Europe over the last decades. PMID- 11880975 TI - Acute changes in gill Na+-K+-ATPase and creatine kinase in response to salinity changes in the euryhaline teleost, tilapia (Oreochromis mossambicus). AB - Some freshwater (FW) teleosts are capable of acclimating to seawater (SW) when challenged; however, the related energetic and physiological consequences are still unclear. This study was conducted to examine the changes in expression of gill Na(+)-K(+)-ATPase and creatine kinase (CK) in tilapia (Oreochromis mossambicus) as the acute responses to transfer from FW to SW. After 24 h in 25 ppt SW, gill Na(+)-K(+)-ATPase activities were higher than those of fish in FW. Fish in 35 ppt SW did not increase gill Na(+)-K(+)-ATPase activities until 1.5 h after transfer, and then the activities were not significantly different from those of fish in 25 ppt SW. Compared to FW, the gill CK activities in 35 ppt SW declined within 1.5 h and afterward dramatically elevated at 2 h, as in 25 ppt SW, but the levels in 35 ppt SW were lower than those in 25 ppt SW. The Western blot of muscle-type CK (MM form) was in high association with the salinity change, showing a pattern of changes similar to that in CK activity; however, levels in 35 ppt SW were higher than those in 25 ppt SW. The activity of Na(+) K(+)-ATPase highly correlated with that of CK in fish gill after transfer from FW to SW, suggesting that phosphocreatine acts as an energy source to meet the osmoregulatory demand during acute transfer. PMID- 11880976 TI - Diverse strategies for ion regulation in fish collected from the ion-poor, acidic Rio Negro. AB - We measured unidirectional ion fluxes of fish collected directly from the Rio Negro, an extremely dilute, acidic blackwater tributary of the Amazon. Kinetic analysis of Na(+) uptake revealed that most species had fairly similar J(max) values, ranging from 1,150 to 1,750 nmol g(-1) h(-1), while K(m) values varied to a greater extent. Three species had K(m) values <33 micromol L(-1), while the rest had K(m) values >or=110 micromol L(-1). Because of the extremely low Na(+) concentration of Rio Negro water, the differences in K(m) values yield very different rates of Na(+) uptake. However, regardless of the rate of Na(+) uptake, measurements of Na(+) efflux show that Na(+) balance was maintained at very low Na(+) levels (<50 micromol L(-1)) by most species. Unlike other species with high K(m) values, the catfish Corydoras julii maintained high rates of Na(+) uptake in dilute waters by having a J(max) value at least 100% higher than the other species. Corydoras julii also demonstrated the ability to modulate kinetic parameters in response to changes in water chemistry. After 2 wk in 2 mmol L(-1) NaCl, J(max) fell >50%, and K(m) dropped about 70%. The unusual acclimatory drop in K(m) may represent a mechanism to ensure high rates of Na(+) uptake on return to dilute water. As well as being tolerant of extremely dilute waters, Rio Negro fish generally were fairly tolerant of low pH. Still, there were significant differences in sensitivity to pH among the species on the basis of degree of stimulation of Na(+) efflux at low pH. There were also differences in sensitivity to low pH of Na(+) uptake, and two species maintained significant rates of uptake even at pH 3.5. When fish were exposed to low pH in Rio Negro water instead of deionized water (with the same concentrations of major ions), the effects of low pH were reduced. This suggests that high concentrations of dissolved organic molecules in the water, which give it its dark tea color, may interact with the branchial epithelium in some protective manner. PMID- 11880977 TI - Nitrogen requirement of the omnivorous greater bilby, Macrotis lagotis (Marsupialia: Peramelidae). AB - Mainly due to their utilisation of relatively low-fiber diets compared to herbivorous mammals, omnivores are expected to have correspondingly low maintenance nitrogen requirements (MNRs). The limited studies examining nitrogen requirements of omnivorous mammals to date have shown this to be the case. In this article, we determine the dietary MNR of greater bilbies (Macrotis lagotis), arid-zone omnivorous marsupials, by feeding them varying proportions of mixed seeds and dried currants (sun-dried grapes). We also examine the possibility that bilbies conserve nitrogen by recycling endogenous urea to their gastrointestinal tract. The dietary MNR of 127 mg N kg(-0.75) d(-1) calculated for the bilby falls within the range calculated for other marsupial omnivores and is lower than that of any of the herbivores. This low requirement for nitrogen was correlated with significant recycling of endogenous urea to the gut; bilbies recycled between 44% and 80% of urea synthesised in the liver, but the proportion recycled was independent of dietary nitrogen intake. The relatively low MNR of the bilby is consistent with its low rates of basal metabolism and the recycling of endogenous urea to the gut, both of which reduce urinary nitrogen losses, and a low-fiber diet that minimises metabolic fecal nitrogen loss. Because the bilby inhabits an environment where its food supply is unpredictable and of seasonally low nitrogen content, this species is likely to benefit from its low requirement for nitrogen. PMID- 11880978 TI - Microclimates and energetics of free-living box turtles, Terrapene carolina, in South Carolina. AB - We measured microclimate, field metabolic rates (FMRs), water flux, and activity patterns of telemetered box turtles (Terrapene carolina) in South Carolina from September 1987 to October 1988. Turtles were inactive for most of the winter and were active only sporadically during the rest of the year. Using the doubly labeled water method, we found that water flux averaged 8.8, 18.9, and 26.4 mL kg(-1) d(-1) in winter, spring, and summer/fall, respectively. FMR for the same periods averaged 0.028, 0.065, and 0.124 mL CO(2) g(-1) h(-1). Differences in FMR among seasons were significant but not between sexes. Using operative temperatures, we predicted standard and maximum metabolic rates of turtles. In winter, FMR was elevated above standard metabolic rates and close to maximum metabolic rates, whereas in spring and summer/fall, FMR fell midway between standard and maximum metabolic rates. We used a model to predict metabolic rates, geographical distribution, and potential reproductive output of box turtles across latitudes in eastern North America. Low FMR and low annual reproductive output may allow box turtles to survive and flourish in unpredictable resource environments by minimizing costs and risks, thereby maintaining greater lifetime reproductive success. PMID- 11880979 TI - Regional blood flow in sea turtles: implications for heat exchange in an aquatic ectotherm. AB - Despite substantial knowledge on thermoregulation in reptiles, the mechanisms involved in heat exchange of sea turtles have not been investigated in detail. We studied blood flow in the front flippers of two green turtles, Chelonia mydas, and four loggerhead turtles, Caretta caretta, using Doppler ultrasound to assess the importance of regional blood flow in temperature regulation. Mean blood flow velocity and heart rate were determined for the water temperature at which the turtles were acclimated (19.3 degrees-22.5 degrees C) and for several experimental water temperatures (17 degrees-32 degrees C) to which the turtles were exposed for a short time. Flipper circulation increased with increasing water temperature, whereas during cooling, flipper circulation was greatly reduced. Heart rate was also positively correlated with water temperature; however, there were large variations between individual heart rate responses. Body temperatures, which were additionally determined for the two green turtles and six loggerhead turtles, increased faster during heating than during cooling. Heating rates were positively correlated with the difference between acclimation and experimental temperature and negatively correlated with body mass. Our data suggest that by varying circulation of the front flippers, turtles are capable of either transporting heat quickly into the body or retaining heat inside the body, depending on the prevailing thermal demands. PMID- 11880980 TI - Ventilation and metabolism in a large semifossorial marsupial: the effect of graded hypoxia and hypercapnia. AB - Metabolic and ventilatory variables were measured in a large semifossorial marsupial, the hairy-nosed wombat (Lasiorhinus latifrons, 21.9 kg). In normoxia, the rate of oxygen consumption was 63% of that predicted for a similar-sized marsupial, and the level of ventilation (V(E)) was such that the convective requirement (V(E)/VO2) was similar to other mammals. Exposure to hypercapnia (5% CO(2)) evoked a hyperventilatory response (3.55 x normoxia) that was no different to that observed for epigeal (surface-dwelling) marsupials; the increase in V(E) was primarily achieved with an increase in tidal volume. Exposure to hypoxia (15% to 8% O(2)) resulted in a hyperventilation (principally through an increase in frequency), although the response was blunted (in 8% O(2), 1.85 x normoxia) and only at the severest levels did hypometabolism contribute. The attenuated response to hypoxia in the wombat is presumably a reflection of a semifossorial lifestyle and a tolerance to this respiratory stimulant. PMID- 11880981 TI - Pulmonary carbonic anhydrase in the garter snake, Thamnophis sirtalis. AB - This study examined whether the snake lung possesses intravascular carbonic anhydrase (CA). Lungs were perfused with control salines and with salines containing CA inhibitors. Perfusion with control salines resulted in a stable CO(2) excretion, whereas CA inhibitors significantly reduced pulmonary CO(2) excretion. Membrane-permeable and membrane-impermeable CA inhibitors produced comparable decreases in CO(2) excretion, suggesting that extracellular, intravascular CA participated in the pulmonary CO(2)-HCO3(-)-H(+) reactions. Treatment of lungs with phosphatidylinositol specific-phospholipase C (PI-PLC) significantly decreased CO(2) excretion, indicating that CA was connected to the luminal endothelial cell membrane by a phosphatidylinositol glycan linkage. Taken together, these results are the first to demonstrate the presence of membrane bound, intravascular CA (CA IV) in the snake lung. PMID- 11880982 TI - Once a fast cod, always a fast cod: maintenance of performance hierarchies despite changing food availability in cod (Gadus morhua). AB - To examine whether Atlantic cod maintain constant hierarchies of sprint speeds and muscle metabolic capacities under different feeding regimes, the physiological capacities of individual cod were followed through a starvation feeding-starvation cycle. We examined sprint speeds and maximal enzyme activities in white-muscle biopsies at each period. We measured the glycolytic enzymes, phosphofructokinase (PFK) and lactate dehydrogenase (LDH), the mitochondrial enzyme, cytochrome C oxidase (CCO), and the biosynthetic enzyme, nucleotide diphosphate kinase (NDPK). Sprint speeds were measured in a laser diode/photocell timed raceway. As expected, the feeding regime had a marked impact on the physiological capacities of cod, but the responses differed for sprint-swimming and muscle metabolic capacities. The different enzyme activities as well the condition index generally decreased during the first starvation, improved with feeding, and fell again during the second starvation. In contrast, sprint performance improved after feeding but did not fall with the second starvation. Although both the enzyme activities and the sprint speeds showed considerable interindividual variation, sprint speeds were not significantly correlated with the enzyme activities. The hierarchy of sprint performance of the cod was maintained, regardless of the preceding feeding regime, whereas those of muscle metabolic capacities were not. PMID- 11880983 TI - Baseline and stress-induced plasma corticosterone during long-distance migration in the bar-tailed godwit, Limosa lapponica. AB - The specific roles of corticosterone in promotion of avian migration remain unclear even though this glucocorticosteroid is elevated in many migrating bird species. In general, glucocorticosteroids promote metabolic homeostasis and may elicit effects on feeding and locomotion. Because the migratory stages of refueling and flight are characterized by distinct behaviors and physiology, the determination of corticosterone levels during each stage should help identify potential processes in which corticosterone is involved. We measured baseline levels of corticosterone in bar-tailed godwits (Limosa lapponica) during two distinct stages of migration: (1) immediately after arrival at a false stopover site just short of the Wadden Sea and (2) throughout the subsequent 4-wk refueling period on the Wadden Sea. Plasma corticosterone was higher in arriving than in refueling birds. In addition, corticosterone increased with size corrected body mass during the refueling phase, suggesting that corticosterone rises as birds prepare to reinitiate flight. Therefore, elevated corticosterone appears associated with migratory flight and may participate in processes characterizing this stage. We also performed a capture stress protocol in all birds and found that corticosterone increased in both arriving and refueling godwits. Therefore, the normal course of migration may be typified by corticosterone concentrations that are lower than those associated with stressful and life-threatening episodes. PMID- 11881028 TI - Gene therapy for hypertension and restenosis. PMID- 11881029 TI - Effectively targetting the renin-angiotensin-aldosterone system in cardiovascular and renal disease: rationale for using angiotensin II receptor blockers in combination with angiotensin-converting enzyme inhibitors. PMID- 11881030 TI - ACE inhibitors and antihypertensive treatment in diabetes: focus on microalbuminuria and macrovascular disease. PMID- 11881031 TI - The renin-angiotensin-aldosterone system and fibrinolysis. AB - Activation of the RAAS has been linked with an increased risk of myocardial infarction and stroke,(1,2,37,38) and recently these beneficial effects have, in part, been attributed to the effects of the RAAS on the fibrinolytic system. Indeed, ACE seems to occupy a central position in modulating the fibrinolytic balance, where an angiotensin II-mediated increase of PAI-1 plays a major role. By contrast, the effect on bradykinin stimulated t-PA release may be of lesser importance, although the data are conflicting. Importantly, the impact of the RAAS on the fibrinolytic balance may also contribute to the favourable effects of ACE inhibition and AT1-receptor antagonists on cardiovascular events, particularly when considering the activation of the RAAS in hypertension and heart failure. More work is clearly required in this area to elucidate potential therapeutic targets. PMID- 11881032 TI - Early initiation of ACE inhibitor treatment after acute myocardial infarction--a missed therapeutic opportunity? PMID- 11881033 TI - Blunted suppression of plasma renin activity in diabetes. AB - We have documented, contrary to expectation, that the renin-angiotensin aldosterone system (RAAS) is stimulated normally by restriction of sodium intake inpatients with Type 2 diabetes mellitus (DM) and hypertension. Conversely, plasma renin activity (PRA)is suppressed less than in normal subjects by a high salt diet in these patients. Increasing plasma angiotensin II (Ang II) concentration through intravenous Ang II infusion also suppresses renin release, via the short feedback loop. In this study, we sought to ascertain whether the limited renin suppression in Type 2 diabetes mellitus via high-salt intake is a unique defect or part of a more generalised abnormality of PRA suppression. We studied 38 patients with Type 2 DM and hypertension, 158 hypertensive control patients, and 61 normotensive controls. All patients were studied while in metabolic balance on a 10 mEq sodium (Na) diet. The response to the Ang II infusion at 3 ng/kg/min for 45 minutes was measured. We found that PRA fell significantly in normal subjects, from 4.0 +/- 0.33 to 2.5 +/- 0.23 ngAngI/ml/hr (p=0.0056). In patients with essential hypertension, the Ang II infusion also led to a fall in PRA from 3.51 +/- 0.23 to 2.76 +/- 0.17 ng Angl/ml/hr(p=0.014). In patients with DM, despite a similar basal PRA (3.7 +/- 0.40 ng AngI/ml/hr), the infusion of Ang II did nor influence PRA significantly (3.43 +/- 0.42ng AngI/ml/hr; p > 0.77), though these patients had the most robust mean arterial pressure response. Our data are in complete accord with the concept of high intrarenal Ang II in DM and suggest lower systemic Ang II despite comparable PRA. PMID- 11881034 TI - Effects of losartan on haemodynamic parameters and angiotensin receptor mRNA levels in rat heart after myocardial infarction. AB - We investigated the haemodynamic parameters and the regulation of cardiac mRNA levels of the angiotensin receptor subtypes, AT1 and AT2, by the AT,-receptor antagonist losartan in rat heart during the acute phase of myocardial infarction. AT1- andAT2-receptor mRNA levels markedly increased at 30 minutes and peaked at 24 hours post myocardial infarction (12.6-fold increase for AT1- and 17.2-fold increase for AT2 compared with controls). Losartan significantly reduced mean blood pressure in sham-operated rats and decreased mean blood pressure and left ventricular end-diastolic pressure in myocardial infarction rats. However, the AT,- andAT2-receptor mRNA levels of losartan-treated rats showed a pattern similar to that of water-treated rats. The time-dependent increase of AT1- and AT2-receptor mRNA levels is associated with the early remodelling process of non infarcted myocardium post MI and is independent of AT1-receptor blockade. PMID- 11881035 TI - The involvement of AT(2)-receptor in the antithrombotic effect of losartan in renal hypertensive rats. AB - In previous studies, we have shown that losartan possesses nitric oxide-dependent antithrombotic properties in various models of hypertension in rats. It was demonstrated that stimulation of AT2-receptors plays an important role in the pharmacological effects of AT1-receptor antagonists. Thus, in this study, we examine the participation of AT2-receptors in the antithrombotic action of losartan in renal hypertensive rats on venous thrombosis induced by a two-hour ligation of the vena cava. Losartan administration(30 mg/kg, p.o.) resulted in a marked decrease in thrombus weight (by 85%, p<0.001). PD123319, an AT2-receptor antagonist (10 mg/kg, i.v.), administered concomitantly with losartan, abolished its antithrombotic effect, whilst it had no influence on thrombus weight when given alone. A significant decrease in systolic blood pressure was observed in animals given losartan. PD123319 administration didnot abolish this action of losartan and did not alter blood pressure when given alone. No changes in prothrombin time, activated partial thromboplastin time, or euglobulin clot lysis time were observed in animals administered losartan and/or PD123319.Similarly, primary haemostatics evaluated by bleeding time and platelet count did not change in any group of rats. In conclusion, we have shown that AT2-receptor stimulation is involved in the antithrombotic action of losartan in renal hypertensive rats. PMID- 11881036 TI - The antithrombotic effect of angiotensin-(1-7) closely resembles that of losartan. AB - Angiotensin-(1-7) [Ang-(1-7)] is the bioactive peptide which may be responsible for some of the pharmacological effects of losartan. Our previous study has demonstrated the antithrombotic action of losartan in a model of experimental thrombosis. In the present study, we compared the antithrombotic action of losartan and Ang-(1-7). Acute (10 mg/kg, p.o.) and chronic (10 mg/kg, p.o., three weeks) losartan administration to spontaneously hypertensive rats (SHR) induced a decrease in thrombus weight (1.6 +/- 0.6 mg and 1.2 +/- 0.3 mg respectively vs. control 2.9 +/- 0.8 mg; p<0.05, p<0.05). A similar reduction was observed in two kidney, one-clip hypertensive rats (2K-IC)receiving acute losartan administration (1.39 +/- 0.29 mg vs. 3.25 +/- 0.62 mg; p<0.01). Infusion of Ang-(1-7) to2K-lC rats also reduced the thrombus weight(1.01 +/- 0.34 mg, 1.23 +/- 0.38 mg and 2.17 +/- 0.36 mg for 1, 10, 100 pmol/kg/min, respectively vs. 3.58 +/- 0.6 mg control; p<0.01, p<0.01, p<0.05). Losartan produced a decrease in systolic blood pressure (BP) in SHR as well as in 2K-1C rats, while Ang-(1-7) infusion had no effect on BP. Acute losartan dosing to 2K-1C rats decreased platelet adhesion to fibrillar collagen(24.9 +/- 1.0% vs. control 31.5 +/- 1.1%, p<0.001). The incubation of platelet samples with Ang-(1-7) (10-6 and 10 5 M) also reduced adhesion to fibrillar collagen(38.4 +/- 0.1% and 33.8 +/- 0.8% respectively vs. control 40.0 +/- 0.6%; p<0.05, p<0.001). There were no apparent changes in prothrombin time, activated partial thromboplastin time and euglobulin clot lysis time in losartan and Ang-(1-7)-treated groups. We conclude that, like losartan, Ang-(1-7) is able to act as an antithrombotic agent. PMID- 11881037 TI - Insulin-like growth factor induces up-regulation of AT(1)-receptor gene expression in vascular smooth muscle cells. AB - BACKGROUND: Insulin-like growth factor-1 (IGF-1), as well as AT1-receptor activation, plays a central role in growth processes of cardiac and vascular cells. In order to assess relevant interactions of both systems, the effect of IGF-1 on AT1-receptor expression was evaluated in vascular smooth muscle cells. METHODS AND RESULTS: Incubation of cultured vascular smooth muscle cells (VSMC) with IGF-1 led to a dose- and time-dependent up-regulation of AT1-receptor mRNA, as measured by Northern hybridisations. The maximal AT1-receptor overexpression of 201 +/- 70% of control levels was reached after a 24-hour incubation with 100 ng/ml IGF-1. Consequently, AT,-receptor protein expression was increased to 231 +/- 35% of control levels. Experiments under transcriptional blockade showed that AT1-receptor mRNA stability was not altered by IGF-1, suggesting that transcriptional mechanisms may be involved in IGF-1-induced AT1-receptor regulation. Preincubation with various pharmacological inhibitors revealed that IGF-1 up-regulated AT1-receptor expression via activation of p42/44 MAP kinase,whereas tyrosine phosphorylation and Pl-3 kinase seemed not to participate in this regulative pathway. CONCLUSIONS: IGF-l-induced up-regulation of the AT1 receptor maybe an important interaction by which cellular grow this modulated in the heart as well as in the vasculature. This may have implications for the treatment regimen of patients suffering from hypertension, cardiac hypertrophy, and coronary heart disease. PMID- 11881038 TI - Losartan reduces collagen content and intimal thickening of iliac arteries after balloon injury in rabbits. AB - Intimal thickening and formation of extracellular matrix are parts of the repair process after vascular injury. Similar processes occur after coronary angioplasty. Prior studies have shown that losartan inhibits intimal thickening in rat carotid arteries following balloon injury. However, the effects of losartan in reducing the collagen content of arteries after balloon injury have not been examined. The objectives of this study were to determine the change in collagen content after balloon injury and to analyse the mechanisms of reduction of collagen content and intimal thickening. Losartan (15 mg/kg/d) was administered orally from six days before to eight weeks after balloon injury in rabbits. Collagen content was measured histologically by the use of circularly polarised images of picrosirius red-stained sections. Collagen content in arterial intima was found to be significantly lower in the losartan-treated group (n= 12) than in the control group (n= 12)(21.6% +/- 5.2% vs. 43.8% +/- 7.6%, p<0.01). Losartanreduced the collagen content in arterial intima by 50.7% area fraction compared with that of control. The morphological observation showed that the intimal area and intimal-to-medial area ratio in the losartan-treated group were significantly less than in the control group (0.27 +/- 0.13mm2 vs. 0.52 +/- 0.29 mm2,0.55 +/- 0.21 vs. 0.97 +/- 0.25, respectively, p<0.05). These data indicate that losartan reduces vascular collagen content and inhibits intimal thickening after balloon injury. The results also suggest that collagen accumulation in the intima may be an important factor in the development of the stenotic lesion and that the use of losartan may have therapeutic value to prevent stenosis after balloon injury. PMID- 11881039 TI - Lys(199) mutation of the human angiotensin type 1 receptor differentially affects the binding of surmountable and insurmountable non-peptide antagonists. AB - Many slow dissociating (insurmountable) non-peptide angiotensin type 1 receptor (AT1) antagonists contain,besides the acidic biphenyltetrazole substructure of losartan, a second acidic group to stabilise antagonist-receptor complexes. To investigate the involved basic amino-acids of the human AT1-receptor, wild-type and mutant receptors were transiently transfected in CHO-K1 cells and characterised by [3H]candesartan binding. Lys199-->Gln substitution decreased the affinity 45-fold for candesartan (95% insurmountable),18-fold for EXP3174 (70% insurmountable), 10-fold for irbesartan (40% insurmountable) and 5-fold for losartan (surmountable). His256 -->Ala substitution had only minor effects. This suggests that Lys199 is important for the tight binding of non-peptide antagonists. PMID- 11881040 TI - The effects of the addition of losartan on uric acid metabolism in patients receiving indapamide. AB - OBJECTIVE: A number of adverse metabolic effects are associated with indapamide administration, including an increase in serum uric acid levels. It has been reported that losartan can significantly decrease serum uric acid levels. However, there are no data on the effects of combination therapy of losartan with indapamide on uric acid metabolism. METHODS: We studied 20 hypertensive patients in whom serum metabolic parameters, including uric acid levels in serum and urine, were studied before and after eight weeks of indapamide administration (2.5 mg once daily) as well as eight weeks after combination treatment with indapamide (2.5 mg once daily)and losartan (50 mg/day). RESULTS: Indapamide evoked a significant decrease in systolic and diastolic blood pressure from a mean value of 157 +/- 12 mmHg/96 +/- 10 mmHg to a mean value of 139 +/- 14 mmHg/92 +/- 5 mmHg (p<0.01 for both comparisons). However, a significant increase in serum uric acid levels was noticed after indapamide administration (from a mean value of 4.9 +/- 1.6 mg/dl to a mean value of 5.9 +/- 1.2 mg/dl, p<0.01), associated with a decrease in the fractional excretion of uric acid(from a mean value of 9 +/- 5% to a mean value of 7 +/- 5.5%, p<0.05). The addition of losartan caused a further decrease in blood pressure from a mean value of 139 +/- 14 mmHg/92 +/- 5 mmHg to a mean value of 120 +/- 15 mmHg/84 +/- 4 mmHg (p<0.01 for both comparisons). This was followed by a significant decrease in serum uric acid levels to 5 +/- 1.1 mg/dl(p<0.01) due to a substantial increase in fractional urate excretion (from 7 +/- 5.5 to 8.7 +/- 6%, p<0.05). CONCLUSION: The addition of losartan could offset the hyperuricaemic effect of indapamide administration. PMID- 11881054 TI - Signal transduction mechanisms of the angiotensin II type AT(1)-receptor: looking beyond the heterotrimeric G protein paradigm. PMID- 11881057 TI - The management of the elderly with hypertension. PMID- 11881058 TI - Salt intake and non-ACE pathways for intrarenal angiotensin II generation in man. AB - Angiotensin-converting enzyme (ACE) plays a crucial role in the generation of angiotensin II (Ang II) via conversion from angiotensin I (Ang I). There has been substantial recent interest in non-ACE pathways of Ang II generation in the heart, large arteries, and the kidney. In the case of the human kidney, studied when in balance on a low-salt diet, the renal haemodynamic response to Ang II antagonists substantially exceeds the renal response to ACE inhibitors (ACE-I), suggesting that about 30-40% of Ang II-generation occurs via non-ACE pathways. In this study, we examined the relative contribution of non-ACE pathways, by comparing the response to candesartan and to captopril at the top of the dose response in normal humans when in balance on a low-salt, as well as a high-salt, diet. As anticipated on a low-salt diet, the increase in renal plasma flow (RPF) in response to candesartan (165+/-14 mL/min/1.73 m2) significantly exceeded the response to captopril (118+/-12 mL/min/1.73 m2; p<0.01). In subjects studied on a high-salt diet, the response to candesartan (97+/-20 mL/min/1.73 m2) also significantly exceeded the response to captopril on the same diet(30+/-15 mL/min/1.73 m2; p<0.01). This remarkable response to candesartan in subjects on a high-salt diet,when compared with the response to captopril,suggests that non-ACE dependent Ang II generation was influenced less than the classical renal pathway with an increase in salt intake, so that the percentage of Ang II generated via the non-ACE pathway rose to the 60-70% range. PMID- 11881060 TI - Effect of valsartan and captopril in rabbit carotid injury. Possible involvement of bradykinin in the antiproliferative action of the renin-angiotensin blockade. AB - The effects of the specific angiotensin II (Ang II) AT1-receptor blocker valsartan on events related to restenosis were investigated in rabbits after common carotid balloon injury. Six animals were given valsartan from two days prior to injury until 14 days post-injury. Three control groups (n=6 in each group) were either sham-operated, untreated or treated with the angiotensin converting enzyme (ACE) inhibitor,captopril. Both ACE inhibition and AT,-receptor blockade had marked effects on plasma levels of endothelin ET1, thromboxane TXB2 and 6-keto-PGF1-alpha. The most dramatic effects on ET, levels were seen in rabbits treated with valsartan, where levels were reduced to values close to those for sham-operated animals (96.85 vs. 86.45 pg/ml). Captopril treatment led to a statistically significant (p<0.01) reduction in ET1 levels compared with untreated animals, but the reduction was only about half that seen with AT1 receptor blockade. TXB2 levels doubled (202.58 vs.413.28 pg/ml) upon arterial injury in control animals but rose by only 20-35% in rabbits treated with captopril (246.45 pg/ml) or valsartan (268.13). In untreated animals, 6-keto-PGF1 alpha levels decreased slightly after injury, but for both the captopril and valsartan groups, there were significant increases in levels of this prostaglandin derivative, effects attributed to the action of bradykinins. Levels were highest in the captopril-treated animals. Valsartan and captopril treatment led to a significant reduction in neointimal thickness and the extent of lumen stenosis compared with untreated animals. Both treatments were effective in reducing neointimal area and significantly (p<0.05)reduced cell proliferation. The differences between treatments can be attributed to the different actions of the agents, as valsartan leaves the AT2-receptor unblocked, while captopril, through inhibition of Ang II synthesis, prevents stimulation of both receptors.A combination of both treatments may be a possible way forward in the clinical prevention of restenosis. PMID- 11881062 TI - Discordant responses to two classes of drugs acting on the renin-angiotensin system. AB - INTRODUCTION: Marked heterogeneity characterises blood pressure (BP)responses to antihypertensive drugs. The efficacy of drugs acting on the renin-angiotensin aldosterone system(RAAS) is predicted (albeit weakly) by plasma renin activity (PRA) and it has been assumed that, within individuals, there would be concordance in efficacy between drugs acting at different sites to block the RAAS. DESIGN: The present study was a randomised, double-blind,two-way, crossover study designed to evaluate intra-individual BP responses to an angiotensin II AT receptor blocker (ARB), candesartan cilexetil, and anangiotensin-converting enzyme inhibitor (ACE-I), lisinopril, and to identify potential phenotypic characteristics of patients' responses to the drugs. METHODS: 92 patients with essential hypertension, (mean systolic/diastolic BP 160/101 mmHg) entered the trial,of whom 76 patients completed both treatments. RESULTS: There was marked heterogeneity in response to the two drugs. 50% of patients responded (fall in diastolic BP of>10 mmHg or achieved diastolic pressure <90 mmHg)to both drugs; 16% were non-responders to both drugs; 20% responded to the ACE-I but not the ARB and 15% responded to the ARB but not to the ACE-I. Individual responses to the two drugs were poorly correlated (for diastolic pressure: r=0.19, p=0.11; for systolic pressure: r=-0.01, p=0.92). For the ACE-I, the fall in both systolic and diastolic BP was related to pre-treatment PRA (for diastolic pressure: r=0.31, p=0.008; for systolic pressure: r=0.24, p=0.04). In the case of the ARB, no relationship between the fall in BP and PRA was observed. These observations suggest that more complex mechanisms may be involved in BP reduction with ARBs than with ACE-I. PMID- 11881063 TI - Comparative antihypertensive and renoprotective effects of telmisartan and lisinopril after long-term treatment in hypertensive diabetic rats. AB - This study compared the cardiovascular and renal effects of long-term telmisartan (3 and 10 mg/kg/day)and lisinopril (10 mg/kg/day) in an animal model combining hypertension and diabetes mellitus. It was a parallel-group study of diabetic, spontaneously hypertensive rats (SHR), treated with control or active treatment for eight months. A non-diabetic SHR control group was run in parallel. Diabetes was induced by streptozotocin (45 mg/kg i.v.) in SHRs aged 9-10 weeks. Animals were treated with telmisartan (3 or 10mg/kg/day), lisinopril (10 mg/kg/day) or vehicle. Plasma glucose levels, blood pressure (BP), and urinary protein and albumin excretion were measured monthly. Telmisartan treatment significantly reduced BP of diabetic SHRs in a dose-dependent manner (p<0.05, low-dose, n= 18; p<0.01, high-dose, n=15). The BP reduction in the lisinopril group was similar to that in the telmisartan 10 mg/kg/day group. Compared with non-diabetic SHRs, untreated diabetic SHRs developed severe proteinuria and albuminuria over the experimental period (p<0.01). In diabetic SHRs, proteinuria and albuminuria were dose-dependently and significantly attenuated by treatment with telmisartan (p<0.01 with the higher dose) and lisinopril (p<0.01). Compared with the untreated diabetic SHRs, cardiac hypertrophy was significantly reduced after treatment with both doses of telmisartan and with lisinopril. Telmisartan, 10 mg/kg/day, but not lisinopril, significantly attenuated the diabetes-induced increase in glomerular volume. In conclusion, telmisartan, 10 mg/kg/day, is at least as beneficial as lisinopril, 10 mg/kg/day, in lowering BP, reducing cardiac hypertrophy and attenuating renal excretion of protein and albumin in this model. PMID- 11881065 TI - The influence of angiotensin-converting enzyme inhibitors on the aorta elastin metabolism in diet-induced hypercholesterolaemia in rabbits. AB - Aortic elastin turnover is significantly accelerated in atherosclerosis, partly because of activation of the renin-angiotensin-aldosterone system caused by hypercholesterolaemia. We postulated that angiotensin-converting enzyme inhibitors (ACE-I) prevent the aortic elastin loss in experimental hypercholesterolaemia. Two doses of ACE-I (captopril, enalapril and quinapril) were used: a dose equivalent to that applied to human subjects and a dose 10 times higher. We found that the increase in serum and aortic elastolytic activity in cholesterol-fed rabbits was prevented by high-dose captopril. The elastin content in aorta homogenates from cholesterol-fed rabbits was significantly decreased. The higher dose of captopril, but no other ACE-I, prevented this decrease in aortic elastin content. In cholesterol-fed rabbits the elastin-bound calcium content was significantly elevated. The higher doses of captopril and enalapril lowered the elastin-bound calcium content. In serum and aortic homogenates of cholesterol-fed rabbits, ACE activity was elevated by 15% and 77%, respectively. Both doses of captopril, enalapril and quinapril prevented this cholesterol-induced increase in serum and aortic ACE activity. We conclude that: 1) administration of captopril at doses 10 times higher than those used in humans prevents hypercholesterolaemia increased aortic elastin loss. 2) higher doses of captopril and enalapril prevent the hypercholesterolaemia-induced increase in aortic elastin-bound calcium. PMID- 11881068 TI - Hypertension due to a giant aldosterone-secreting adenoma. PMID- 11881101 TI - Pharmacological properties of angiotensin II antagonists: examining all the therapeutic implications. AB - Angiotensin II (Ang II), the effector peptide of the renin-angiotensin system (RAS), exerts a variety of actions in physiological blood pressure and body fluid regulation, and is implicated as a major pathogenic factor in the development of cardiovascular disease. Inhibition of the RAS, via treatment with the angiotensin converting enzyme inhibitors (ACE-I), or more recently the Ang II AT(1)-receptor blockers (ARBs), has been used as a therapeutic approach to the treatment of hypertension and other cardiovascular dysfunction. Evidence from animal and clinical studies shows that the antihypertensive and overall organ-protective actions of the ARBs are similar to those of ACE-I. However, as the ARBs selectively block the AT(1)-receptor, which is responsible for the known cardiovascular actions of Ang II, leave the AT(2)-receptor unopposed and do not interfere with the breakdown of bradykinin, there is the potential for the beneficial effects in hypertensive patients with cardiovascular diseases such as left ventricular hypertrophy. Furthermore, there may be additional benefits when the ARBs are combined with ACE-I in such patients. Animal studies contribute to the elucidation and understanding of the role of AT(1)- and AT(2)-receptors in the cardiovascular system, and may help in the design of clinical studies aimed at investigating the effects of ACE-I, ARBs, and their combination, on cardiovascular outcomes in hypertensive patients. PMID- 11881103 TI - Angiotensin II receptor blockers and cardiovascular outcomes: the evidence now and in the future. AB - The blood-pressure-lowering efficacy of both angiotensin-converting enzyme inhibitors (ACE-I) and angiotensin II receptor blockers (ARBs) has been clearly demonstrated in recent years, although there is evidence that within the ARB class the individual therapies not necessarily identical in terms of sustained and consistent antihypertensive efficacy over the entire closing period. However, the results of the recent HOPE study have demonstrated that ACE-I have a wider role to play in treating cardiovascular disease, and support the idea that ACE inhibition specifically has a vascular protective effect. the most dramatic benefits were seen in patients with systolic blood pressures in the hypertensive range. The ability of the ACE-I to provide protective effects beyond blood pressure control may be due to their ability to attenuate the breakdown of kinins as well as a role in reducing angiotensin II. These data post the question as to whether the pharmacological properties of the ARBs, in addition to their antihypertensive efficacy, may also play a significant role in influencing cardiovascular outcomes. A number of prospective long-term studies, including VALUE, SCOPE, LIFE, VALIANT, OPTIMAAL, VAL-HEFT and CHARM I-III, are investigating the effects of the ARBs or mortality and morbidity in patients with cardiovascular disease. These studies should answer important questions with respect to the role that ARBs may have in influencing cardiovascular outcomes, although it remains to be seen whether ARBs can match the protective effects of ramipril in high-risk patients. Given the excellent tolerability of the ARBs, it will be of value to examine the influence of ARBs on cardiovascular outcomes in all relevant patient groups. PMID- 11881102 TI - Effect of telmisartan on arterial distensibility and central blood pressure in patients with mild to moderate hypertension and Type 2 diabetes mellitus. AB - Arterial wall stiffness is an important independent risk factor for cardiovascular disease in hypertensive patients, which is further exacerbated by co-existent diabetes mellitus. Increased arterial stiffness is directly associated with an increase in pulse wave velocity (PWV) and indirectly with increased central and peripheral blood pressure. Following a two-week placebo run in period, 27 patients with mild to moderate essential hypertension and Type 2 diabetes mellitus, were randomised to once daily treatment with either telmisartan 40 mg or placebo for three weeks, and after a two-week washout period, crossed-over to the alternative treatment for a further three weeks. Carotid/femoral and carotid/radial PWV were measured non-invasively using the automatic Complior device, and central parameters (central blood pressure, pulse contour analysis, and augmentation index) were measured using the SphygmoCor system, at the start and end of each treatment period. Compared with placebo, treatment with telmisartan significantly reduced carotid/femoral PWV (mean adjusted treatment difference -0.95 m/s, 95% confidence intervals: -1.67, -0.23 m/s, p=0.013), as well as peripheral and central diastolic, systolic and pulse pressure. In conclusion, the results of the this study show that telmisartan is effective in reducing arterial stiffness in hypertensive patients with Type 2 diabetes mellitus, and may potentially have beneficial effects on cardiovascular outcomes, beyond blood-pressure lowering effects in the patient group. PMID- 11881104 TI - Angiotensin II receptor antagonists for the treatment of heart failure: what is their place after ELITE-II and Val-HeFT? PMID- 11881105 TI - Salt - The DASH-Sodium trial. PMID- 11881106 TI - The renin-angiotensin system in the brain: an update. PMID- 11881107 TI - Non-AT(1)-receptor-mediated protective effect of angiotensin against acute ischaemic stroke in the gerbil. AB - Previous studies have shown that angiotensin II (Ang II), by mediating rapid recruitment of collateral circulation, has a protective effect in the setting of acute ischaemia. In an experimental model of acute cerebral ischaemia in the gerbil, Fernandez et al. have reported that the mechanism of the protective effect of Ang 11 is blood pressure (BP)-independent, and that the AT1-receptor antagonist, losartan, but not the ACE inhibitor (ACE-I),enalapril, decreases mortality following unilateral carotid artery ligation. The aim of this study was to examine there producibility of the respective effects of losartan and enalapril, and to verify that these differential effects are drug class-related. Acute cerebral ischaemia was induced in anaesthetised gerbils bv unilateral carotid ligation. The effect of pretreatment with two different ACE-I(enalapril and lisinopril), and two different AT1-receptor antagonists (losartan and candesartan), administered orally or intravenously, on mortality were compared. Kaplan-Meier survival curves at day three were analysed bv a log-rank test. Pretreatment with both enalapril and lisinopril significantly decreased survival at day three compared with controls, while the AT1-receptor antagonists losartan and candesartan, despite similarly lowering BP, did not increase mortality. Coadministration of losartan and enalapril increased mortality to the same extent as enalapril alone. This study confirms that Ang II contributes to protective mechanisms against acute cerebral ischaemia through non AT1-receptor-mediated, BP independent effects. PMID- 11881109 TI - Effects of losartan treatment on T-cell activities and plasma leptin concentrations in primary hypertension. AB - Recent evidence shows that leptin may contribute to elevated blood pressure (BP) and interact with the renin-angiotensin-aldosterone and cellular immune systems. Altered T-cell activities and changes in T-cell subset ratios have also been reported in hypertension. However, little is known about the effects of AT1 receptor antagonism on T-cell activities and plasma leptin concentrations in primary hypertension. We have, therefore, investigated the relationship between leptin and T-cell activities and the effect of an AT1-receptor antagonist, losartan, in primary hypertension. Twenty recently-diagnosed and untreated young adults (11 males and 9 females, age; 39.9+/-7.6 years, range 23-49 years, BMI; 27.6+/-3.7kg/m2) and 20 normotensive healthy, age-, sex- and BMI-matched controls were studied. The [3H]-thymidine uptakes of cultured lymphocytes were determined, both spontaneously and after stimulation with phytohaemagglutinin. The tests were performed before and after three months of treatment with losartan. The results indicate that the blastogenic responses of T-cells to phytohaemagglutinin are significantly higher in the patient group compared with controls (p=0.02). After normalisation of BP, T-cell responses were significantly reduced and were lower than in the controls (p=0.01). Pretreatment plasma leptin levels were significantly higher in hypertensives than in controls (p=0.01). However, losartan treatment had no significant effect on leptin concentrations; moreover, no correlation between leptin levels and T-cell activity was found. Our data show that plasma leptin levels and T-cell activity are markedly enhanced in untreated essential hypertension and that the alteration of T-cell activity is not related to plasma leptin levels. Antihypertensive treatment with losartan decreases T cell activities but does not influence plasma leptin levels. We conclude that leptin levels are not affected by AT1-receptor blockade and are not related to T cell activity. PMID- 11881108 TI - Effects of combination of low doses of angiotensin-converting enzyme inhibitor and diuretics on renal function in spontaneously hypertensive rats: comparison between acute and chronic treatment. AB - The goal of this study was to assess the effect of acute or chronic treatment with S5590, a combination of the angiotensin-converting enzyme inhibitor perindopril (0.76 mg/kg/day) and the diuretic indapamide (0.24 mg/kg/day) on renal function in spontaneously hypertensive rats with moderate renal injury. Renal function was evaluated in conscious rats by clearance methods using labelled inulin and PAH, after catheterisation of the carotid artery, jugular vein and bladder. Both acute and chronic treatment normalised renal vascular resistance, although the effect on blood pressure was more marked after chronic than after acute treatment. Although acute treatment with S5590 increased glomerular filtration rate and renal blood flow, chronic treatment did not affect these parameters. Diuresis and natriuresis were only slightly modified and the results suggest a marked renal vasodilatation. In conclusion, the maintenance of renal function after chronic treatment, in a setting of normalisation of arterial pressure, suggest that such a combined treatment may exert marked renal functional protective effects in hypertension. PMID- 11881110 TI - Angiotensin II-induced stimulation of collagen secretion and production in cardiac fibroblasts is mediated via angiotensin II subtype 1 receptors. AB - The possible contributions of the angiotensin receptor subtypes 1 (AT1) and 2 (AT2) to angiotensin II (Ang II)-induced changes in collagen secretion and production were studied using the specific angiotensin AT1- and AT2-receptor antagonists telmisartan and P-186, respectively. Cardiac fibroblasts (from normal male adult rats) from passage 2 were cultured to confluency and incubated in the presence of 10(-10) to 10(-6) M Ang II in serum-free Dulbecco's MEM medium for 24 hours. Collagen production and secretion were assayed by'H-Proline incorporation; non-collagen production and secretion were also calculated. Ang II dose dependently increased collagen secretion and production in rat adult cardiac fibroblasts in culture. Non-collagen secretion and production were also concentration-dependently increased by Ang II. Addition of 100 nmol/l Ang II increased (p<0.01) collagen secretion and production bv 75+/-6 (SEM)% and 113+/ 23%, respectively, and non-collagen secretion and production by 65+/-6% and 57+/ 16%, respectively. Pretreatment of cardiac fibroblasts with telmisartan completely blocked the Ang II-induced increase in collagen secretion (p<0.001) and production(p<0.05) and in non-collagen secretion (p<0.01) and production (p<0.01). P-186 had no effect on the Ang II-induced increase in collagen secretion and production. Addition of telmisartan and P-186 did not affect collagen secretion and production in basal cardiac fibroblasts. Our data demonstrate that the effects of Ang II on collagen secretion and production in adult rat cardiac fibroblasts in culture are AT1-receptor mediated, since they were abolished by the specific AT1-receptor antagonist, telmisartan, but not by the specific AT2-receptor antagonist, P-186. PMID- 11881111 TI - Effects of telmisartan, hydrochlorothiazide and their combination on blood pressure and renal excretory parameters in spontaneously hypertensive rats. AB - The effects of telmisartan and hydrochlorothiazide (HCTZ) alone and in combination on blood pressure (BP) and renal excretory function were investigated in male spontaneously hypertensive rats (SHR) after oral administration for five consecutive days. Four treatments were studied: vehicle (0.5% Natrosol), telmisartan 3 mg/kg, HCTZ 10 mg/kg, and telmisartan 3 mg/kg+HCTZ 10 mg/kg. The effects on BP and heart rate were studied in 40 SHRs (10 animals per group) using an implanted telemetry device. Renal excretory function was assessed in 76 SHRs (18 animals per group, of which nine were used for urine sampling and nine for blood sampling). The telmisartan/HCTZ combination produced the greatest reductions in trough DBP (-44+/-1.5 mmHg), SBP (-60+/-1.9 mmHg) and mean BP (mBP; -53+/-1.7 mmHg) after five days of therapy (p<0.05 vs. vehicle and vs. telmisartan). Telmisartan monotherapy also decreased DBP, SBP and mBP significantly (p<0.05), but only minor BP fluctuations occurred in SHRs receiving HCTZ or vehicle. Telmisartan/HCTZ elevated heart rate by approximately 12 beats per minute (p<0.05 vs.control). Significant increases in urine volume and Na+, Cl, K+, creatinine and glucose excretion were observed with HCTZ treatment (p<0.01). Telmisartan/HCTZ also promoted renal water and electrolyte excretion (p<0.01); the diuretic effect appeared to be greater with the combination than with HCTZ alone, and there was some attenuation of urinary K+ loss. Elevated blood urea nitrogen levels were observed only in HCTZ-treated SHRs. These results indicate that the antihypertensive efficacy of telmisartan in SHRs is augmented by co-administration with HCTZ. The combination did not affect renal excretory function, with the exception of an increase in blood urea nitrogen and a possible amelioration of HCTZ-related K+ depletion. PMID- 11881112 TI - Effects of different durations of pretreatment with losartan on myocardial infarct size, endothelial function, and vascular endothelial growth factor. AB - A previous study by our group showed that 10 weeks of pretreatment with losartan reduced myocardial infarct size and arrhythmias in a rat model of ischaemia reperfusion. However, the effect of a differing time course of pretreatment has not been investigated. 104 Sprague-Dawley rats were randomised to four groups: a control, and three treatment groups in which losartan (40 mg/kg/day) was administered in drinking water for one day, one week, and four weeks respectively. After different durations of pretreatment, the rats were subjected to 17 minutes of left coronary artery occlusion and 120 minutes of reperfusion. Haemodynamic variables were not significantly different between the four groups. Myocardial infarct size was unchanged after one day and one week of pretreatment (52+/-7, 57+/-6% vs.control 55+/-3%), but was significantly reduced by four weeks of pretreatment with losartan (38+/-6, p<0.05). Endothelial-dependent vasorelaxation was significantly increased by four weeks of pretreatment (-81+/-4 vs.-62+7%, p<0.05). As an indicator of ischaemia, vascular endothelial growth factor (VEGF) levels in ischaemic myocardium were decreased after one and four weeks of pretreatment (0.75+/-0.05, 0.58+/-0.10 vs. 1.0, p<0.05,0.01, respectively). In conclusion, losartan has time-dependent cardiovascular protective effects. Four weeks of pretreatment with losartan decreased infarct size and VEGF, and improved endothelial dysfunction. PMID- 11881113 TI - Enhanced regional AT(2)-receptor and PKC(epsilon) expression during cardioprotection induced by AT(1)-receptor blockade after reperfused myocardial infarction. AB - We assessed the effects of the angiotensin II (Ang II) type 1 receptor (AT1 receptor) blocker, candesartan, (CN, 1 mg/kg i.v. over 30 minutes pre-ischaemia) alone or after intracoronary administration of Ang II type 2 receptor (AT2 receptor) blocker (PD 123319), protein kinase C (PKC) inhibitor (chelerythrine), endothelial nitric oxide (NO) synthase inhibitor (N(G)-monomethyl-L-arginine or L NMMA), and bradykinin (BK) -B2 receptor inhibitor (HOE140) on in vivo left ventricular (LV) function and remodelling (echocardiograms/Doppler) and haemodynamics in 30 dogs with reperfused anterior infarction (90 minutes ischaemia, 120 minutes reperfusion), and ex vivo infarct size, AT1-receptor/AT2 receptor proteins and PKC(epsilon) (immunoblots), and cyclic guanosine 3', 5' monophosphate (cGMP, immunoassay). Compared with controls, CN inhibited the Ang II pressor response, reduced LV preload, improved LV systolic and diastolic function, limited LV remodelling, decreased infarct size, and increased AT2 receptor and PKC(epsilon) proteins in the infarct zone (IZ), and these responses were abrogated by PD 123319, chelerythrine, L-NMMA and HOE140. In addition, the increase in LV cGMP with CN was attenuated by PD 123319, L-NMMA and HOE140. The overall results suggest that AT2-receptor activation and signalling via BK, PKC(epsilon) and cGMP contribute to cardioprotection associated with AT1-receptor blockade during ischaemia-reperfusion injury. PMID- 11881115 TI - Inhibition of the renin-angiotensin system, with particular reference to dual blockade treatment. PMID- 11881114 TI - Valsartan and candesartan can inhibit deteriorating effects of angiotensin II on coronary endothelial function. AB - The angiotensin II (Ang II) AT1-receptor antagonists, valsartan and candesartan, were compared with regard to their effect on Ang II-mediated changes in parameters of coronary endothelial function. Ang II (10 microM) induced increased concentrations of the vasoconstrictor endothelin, the procoagulatory substance plasminogen-activator-inhibitor-1 (PAI-1) and the precursor of the matrix metalloproteinase 1 (MMP-1) in endothelial cell cultures from human coronary arteries. These increases were completely prevented by the addition of 10 microM valsartan or candesartan and partially by the addition of lower concentrations of these drugs, i.e. 1 microM and 0.1 microM. No significant difference between the effect of the two AT1-receptor antagonists was observed. These results suggest that AT1-receptor antagonists not only can reduce blood pressure by blocking the action of Ang II, but might also contribute to the prevention of atherogenesis and plaque instability. PMID- 11881116 TI - Blood pressure and stroke; the PROGRESS trial. PMID- 11881117 TI - Diagnosis and management of primary aldosteronism. PMID- 11881118 TI - Renoprotection in Type 2 diabetes: blockade of the renin-angiotensin system with angiotensin II receptor blockers. PMID- 11881120 TI - Contractile effects of angiotensin peptides in rat aorta are differentially dependent on tyrosine kinase activity. AB - It has been suggested that tyrosine kinase activity participates in the regulation of signal transduction associated with angiotensin II (Ang II)-induced pharmaco-mechanical coupling in rat aortic smooth muscle. We further tested the effects of genistein, a tyrosine-kinase inhibitor, and its inactive analogue, daidzein, on angiotensin I (Ang I), angiotensin III (Ang III) and angiotensin IV (Ang IV) contractions, as compared with those on Ang II. Genistein partially inhibited Ang II- and Ang I-induced contractions. The genistein-induced inhibition was more evident on Ang III and especially important on Ang IV contractile effects. Thus, Ang IV- and Ang III-induced contractions seem to be more dependent on tyrosine kinase activity than those evoked by Ang II or Ang I. Daidzein did not significantly affect the contractile effects of any of angiotensin peptides tested. These results clearly suggest that the inhibition of the action of angiotensin peptides actions by genistein is mediated by inhibition of endogenous tyrosine kinase activity. Furthermore, our data show that the type and/or intensity of tyrosine kinase activity is differentially associated with the contractile effects of different angiotensin peptides in rat aorta. Nifedipine, a blocker of membrane L-type Ca2+ channels, strongly inhibited Ang IV induced contractions. At the same time, it significantly inhibited Ang III contractile effects as compared with Ang II and Ang I contractions. Meanwhile, we observed a close relationship between calcium influx and tyrosine kinase phosphorylation activity under the stimulatory effects of angiotensin peptides. Furthermore, genistein did not significantly influence the phasic contractions induced by angiotensin peptides in Ca2+-free Krebs-Henseleit solution. Thus, it appears that Ca2+ influx, rather than the release of Ca2+ from IP3-sensitive stores, may play a major role in the contractile effects of angiotensin peptides in rat aorta via tyrosine kinase activation. One argument against a direct action of genistein on the Ca2+ channel itself is that it did not markedly affect the K+ induced contraction (depolarisation) in rat aorta. At the same time, a potential role for tyrosine kinase activity in the process of calcium entry is suggested. An elevation of intracellular calcium via tyrosine kinase-mediated processes may mediate the actions of G-protein coupled receptor agonists in smooth muscle, including angiotensin peptides. PMID- 11881119 TI - Pre-treatment with candesartan protects from cerebral ischaemia. AB - Angiotensin II (Ang II) regulates cerebral blood flow by stimulating cerebral vasoconstriction via AT1-receptors. In adult spontaneously hypertensive rats (SHR), the cerebrovascular autoregulatory curve is shifted to the right, in the direction of higher blood pressures, an indication of excessive cerebrovascular vasoconstriction. A restricted capacity to dilate cerebral blood vessels may be responsible for the enhanced vulnerability to cerebrovascular ischaemia during hypertension. We found that chronic treatment with the AT1-receptor antagonist, candesartan, (0.5 mg/kg/day for 14 days, via osmotic minipumps implanted in the subcutaneous tissue) blocked Ang II binding to AT1-receptors in cerebral blood vessels and in brain areas involved in the regulation of cerebrovascular flow, and increased the ratio of lumen-wall area in the middle cerebral artery. Candesartan treatment normalised the lower part of the autoregulatory curve in SHR, and markedly decreased cerebral ischaemia as a consequence of middle cerebral artery occlusion with reperfusion. Protection from ischaemia is related to arterial remodelling, enhanced compensatory vasodilatation in the peripheral area of ischaemia, decreased reduction in cerebral blood flow following the occlusion of a major cerebral blood vessel, and protection from injury in the periphery of the lesion. Our results indicate that pre-treatment with AT1 antagonists such as candesartan could be of benefit in the prevention and treatment of brain ischaemia. PMID- 11881121 TI - No effect of angiotensin II AT(2)-receptor antagonist PD 123319 on cerebral blood flow autoregulation. AB - Blockade of the renin-angiotensin system with angiotensin-converting enzyme inhibitors (ACE-I) or angiotensin AT1-receptor antagonists shift the limits of autoregulation of cerebral blood flow (CBF) towards lower blood pressure (BP). The role of AT2-receptors in the regulation of the cerebral circulation is uncertain. Hence, the present study investigated the effect on CBF autoregulation of blocking of angiotensin AT2-receptors with PD 123319 in spontaneously hypertensive rats (SHR). Anaesthetised and ventilated SHR were given PD 123319, 0.36 mg/kg/min, intravenously, and compared with a control group. CBF was measured by the intracarotid 133xenon injection method and BP was raised by noradrenaline infusion and lowered by controlled haemorrhage in separate groups of rats. The limits of autoregulation were determined by computed least-sum-of squares analysis. PD 123319 did not influence baseline CBF, but resulted in a minor BP decrease (10 control and 10 treated rats). The lower limit of CBF autoregulation (eight treated and eight control) as well as the upper limit of CBF autoregulation (eight treated and eight control) were not significantly different in PD 123319 and control animals (lower limit treated 102+/-4 mmHg and control 94+/-4; NS, and upper limit treated 171 +/- 10 mmHg and control 162+/-7; NS). These findings indicate that acute AT2-receptor blockade does not influence CBF autoregularion. PMID- 11881122 TI - Increased serum angiotensin-converting enzyme activity and plasma angiotensin II levels during pregnancy and postpartum in the diabetic rat. AB - OBJECTIVE: The renin-angiotensin system (RAS) plays an important role in the regulation of blood pressure, electrolyte balance and renal function in normal human pregnancy. The present study was designed to assess various components of the RAS and renal function during pregnancy and immediately after pregnancy in the streptozotocin (STZ)-diabetic rat. METHODS: Pregnant Wistar rats were allocated to three groups: I-control, non-diabetic rats (n=24), II-STZ-diabetic rats (STZ 55 mg/kg body weight, i.v. on day 10 of pregnancy, n=24), III-diabetic rats, as above, treated with insulin (4 units/day, s.c. n=21). On days 17-18 of pregnancy, or within 24 hours after delivery, the rats were sacrificed and the various components of the RAS were determined. RESULTS: Urinary protein excretion (UP) and creatinine clearance(CCr) were greater in group II, four days after STZ, than in group I (UP: I-7.6+/-2.8, II-18.6+/-6.3 mg/24-hour, p<0.001, CCr: I 1.04+/-0.33, II-2.38+/-0.7 ml/minute, p<0.001). Mean (+/-SD) serum angiotensin converting enzyme (ACE) activity and plasma angiotensin II(Ang II) levels at days 17-18 of pregnancy were greater in the untreated diabetic rats than in control pregnant rats (ACE: 163+/-18 vs. 111+/-21 nmol/ml/minute, p<0.001, Ang II: 115+/ 45 vs. 43+/-10 pg/ml, p<0.005). Postpartum serum ACE activity and plasma Ang II levels were greater in group II (ACE: I-123+/-14, II-142+/-24, III-108+/-21 nmol/ml/minute, p<0.01, Ang II: I-56+/-38, II-148+/-62, III-38+/-17 pg/mI, p<0.001). ACE activity in the lung was greater, whereas the activity in the renal cortex was less, in group II than in group I. Kidney weight in untreated diabetic rats was greater than in the other two groups. CONCLUSION: Increased serum ACE activity during pregnancy and postpartum in the untreated diabetic rat is associated with enhanced serum Ang II levels, which may contribute to increased protein excretion and renal hypertrophy. PMID- 11881124 TI - Bioactive angiotensin peptides: focus on angiotensin IV. PMID- 11881123 TI - The effects of candesartan on vascular responses to angiotensin II and norepinephrine in normal volunteers. AB - The effects of chronic administration of candesartan, 16 mg once-daily, to normal volunteers on cardiovascular responses to angiotensin II (Ang II) and norepinephrine (NE) were examined. Fifteen healthy, non-smoking volunteers participated in a randomised, double-blind crossover study of two weeks of candesartan therapy, compared with two weeks of placebo. Blood pressure (BP) responses were measured to increasing infusion rates of intravenous Ang II and NE, along with forearm blood flow (FBF) responses into intra-brachial arterial Ang II, 2 and 24 hours after the last dose of candesartan or placebo. FBF responses to intra-brachial arterial NE were recorded approximately 2 hours following the final dose. Systolic and diastolic BP responses to intravenous infusions of Ang II during candesartan treatment were completely suppressed and significantly lower than during placebo treatment, 2 hours (candesartan 96+/ 10/55+/-8 mmHg; placebo 105+/-5/64+/-8 mmHg) and 24 hours (candesartan 94+/ 8/54+/-8 mmHg; placebo 103+/-7/64+/-7 mmHg) following the last dose. In contrast, FBF responses to intra-brachial arterial Ang II were significantly suppressed by candesartan compared with placebo in a subgroup of subjects 2 hours following the last dose (n=9), but not 24 hours after the last dose (n=8). FBF responses to NE were also suppressed by candesartan treatment 2 hours following the last dose, while BP responses to intravenous NE were unaltered. Chronic candesartan therapy, 16 mg once-daily effectively suppresses pressor responses to Ang II over the duration of the dosing interval. PMID- 11881126 TI - Prospects for ARB in the next five years. PMID- 11881125 TI - Aldosterone-induced vasculopathy: a new reversible cause of cardiac death. PMID- 11881127 TI - Intracellular angiotensin II: from myth to reality? PMID- 11881128 TI - Effects of candesartan on cardiac and arterial structure and function in hypertensive subjects. AB - OBJECTIVES: To study the effect of candesartan cilexetil on left ventricular mass index (LVMI), left ventricular systolic and diastolic function, arterial structure and function and blood pressure (BP) in hypertensive patients. DESIGN AND METHODS: Patients (n=35), aged >20 years, with hypertension and average baseline LVMI of 89 g/m2 were treated for 24 weeks with candesartan, 16 mg o.d., following a four-week placebo run-in period. If diastolic BP remained above 95 mmHg, hydrochlorothiazide, 12.5 mg o.d.,was added. Left ventricular structure and function were assessed using transthoracic echocardiography. Arterial function and structure were assessed using pulse wave analysis to calculate augmentation index (AIx) and forearm plethysmography to calculate minimum vascular resistance. BP was measured in the office and by 24-hour ambulatory BP monitoring (ABPM). RESULTS: The mean reduction in LVMI was 4.4 g/m2(p=0.022). Left ventricular systolic function was not significantly altered from baseline, but diastolic function significantly improved: the mean change in diastolic time was 54 ms (p=0.037), in peak velocity filling 6.3 cm/s (p=0.023); E:A ratio improved by 0.08 (p=0.049). The mean reduction in forearm vascular resistance was 15 units at rest (p=0.001) and 1.3 units after limb ischaemia (p=0.006). AIx decreased significantly, with a mean reduction of 9% (p<0.001). Central BP also significantly reduced(systolic blood pressure/diastolic blood pressure 31/20 mmHg; p<0.001). BP was significantly reduced, both in the office (22/16 mmHg; p<0.001) and by 24-hourABPM (18/12 mmHg; p<0.001). CONCLUSIONS: Treatment with candesartan, 16 mg o.d., with or without hydrochlorothiazide, for 24 weeks, significantly reduced left ventricular mass and arterial hypertrophy in patients with hypertension. In parallel, there were significant improvements in left ventricular diastolic function and arterial function. PMID- 11881129 TI - Renin-angiotensin blockade improves renal cGMP production via non-AT(2)-receptor mediated mechanisms in hypertension-induced by chronic NOS inhibition in rat. AB - BACKGROUND: To investigate the changes in the angiotensin II (Ang II) receptors and nitric oxide (NO)-cGMP pathway in the rat kidney after nitric oxide synthase (NOS)blockade. METHODS: Captopril, an angiotensin-converting enzyme (ACE)inhibitor, 20 mg/100 ml; and/or L-158,809 (an Ang II AT1-receptor antagonist, 5 mg/100 ml) and L-NAME (NOS inhibitor, 50 mg/100 ml) were administered orally for 12 weeks. Blood pressure (BP),urinary albumin, urinary cGMP excretion, plasma ANP, and plasma renin activity were measured. In vitro autoradiography was used to locate the Ang II receptors in the kidney. RESULTS: Captopril and L- 158,809 treatments normalised BP and prevented the appearance of albuminuria in rats receiving L-NAME. Urinary cGMP excretion was significantly increased in L-158,809-treated rats compared with the non-treated group, suggesting that the dysfunctional NO system may be activated by the treatment. AT1-receptor binding in the kidney was inhibited to about 40% of the control value after administration of L- 158,809. The AT2-receptor binding was inhibited to less than 15% of the control value. NOS inhibition had no effect on receptor binding. CONCLUSION: Blockade of NOS causes hypertension and renal damage. Treatment with an ACE inhibitor and/or Ang II receptor antagonist prevented these changes equally effectively. The stimulatory effect of AT1-receptor antagonism on cGMP production was not mediated by AT2-receptor-dependent mechanisms, since renalAT2-receptor binding density was suppressed following treatment with L 158,809. AT1-receptor blockade per se favours activation of humoral pathways that stimulate cGMP production potentially contributing to renal and vascular protection in hypertension and chronic renal disease. PMID- 11881130 TI - High serum enalaprilat in chronic renal failure. AB - BACKGROUND: Most angiotensin-converting enzyme (ACE) inhibitors and their metabolites are excreted renally and doses should hence be reduced in renal insufficiency. We studied whether the dosage of enalapril in daily clinical practice is associated with drug accumulation of enalaprilat in chronic renal failure. METHODS: Fifty nine out-patients with plasma creatinine >150 micromol/L and chronic antihypertensive treatment with enalapril were investigated, in a cross-sectional design. RESULTS: Median glomerular filtration rate (GFR) was 23(range 6-60) ml/minute/1.73 m2. The daily dose of enalapril was 10 (2.5-20) mg and the trough serum concentration of enalaprilat was 31.8 (<2.5-584.7)ng/ml. Ninety percent of the patients had higher serum concentrations of enalaprilat than has been reported in subjects with normal kidney function, and a marked elevation of serum enalaprilat was observed in patients with GFR <30 ml/minute. All but three patients had serum ACE activity below the reference range. The ACE genotype did not influence the results. Additional pharmacokinetic studies were done in nine patients in whom GFR was 23 (10-42)ml/minute/1.73 m2. The median clearance of enalaprilat was 28 (16-68) ml/minute and correlated linearly with GFR (r=0.86, p=0.003). Intra-subject day-to-day variation in trough concentrations was 19.7%. CONCLUSION: Patients with chronic renal failure given small or moderately high doses of enalapril may thus have markedly elevated levels of serum enalaprilat. Whether this affords extra renoprotection, or on the contrary may inappropriately impair renal function, is not known, and should be investigated in prospective, controlled studies. PMID- 11881131 TI - Evaluation of the safety and efficacy of telmisartan and enalapril, with the potential addition of frusemide, in moderate-renal failure patients with mild-to moderate hypertension. AB - The effect on renal function and efficacy of the angiotensin II AT1-receptor blocker (ARB), telmisartan, were compared with those of the angiotensin converting enzyme inhibitor, enalapril, for the treatment of mild-to-moderate hypertension (diastolic blood pressure [DBP] 95-114 mmHg) in the presence of moderate renal failure (creatinine clearance [Ccr] 30-80 ml/minute). The study was multicentre, double-blind, double-dummy and active-controlled in design, with patients randomised in a 2:1 ratio to receive telmisartanor enalapril. After a two-week placebo run-in period, the 71 eligible patients received either telmisartan, 40 mg, orenalapril, 10 mg, once-daily for four weeks. Thereafter, doses were titrated to telmisartan 80 mg or enalapril 20 mg once-daily if supine trough DBP was still > or =90 mmHg. After a further four weeks, dose titration was again performed, as required, to telmisartan, 80 mg,or enalapril, 20 mg, or frusemide was given in addition if the double dose was already being administered. Mean Ccr decreases of 4.6% for telmisartan and 2.8% forenalapril were not clinically significant. Adverse events occurred in 12 (26.7%) telmisartan-treated patients and in 12 (46.2%) patients receiving enalapril. The mean reduction in supine trough DBP from baseline to the last available value was 12.5 mmHg for telmisartan,compared with 11.9 mmHg for enalapril. A full (reduction of >or=10 mmHg) or partial (reduction of 7-9 mmHg) response occurred in 78% of telmisartanpatients and 65% of enalapril patients. In the enalapril group, 43% of patients required frusemide, compared with 29% of those in the telmisartan group. In conclusion, telmisartan lacks detrimental effect on renal function, is effective in the treatment of mild-to-moderate hypertension in patients with moderate renal failure,and is comparable to enalapril. PMID- 11881132 TI - Losartan may modulate erythropoietin production. AB - Erythropoietin (Epo) is distinct amongst haematopoietic hormones, in that it is produced remote from the bone marrow. The tissue oxygen pressure required to trigger the Epo gene under physiological conditions is uniquely sited at a restricted area in the kidney termed the critmeter. Angiotensin II (Ang II) increases sodium reabsorption and hence oxygen consumption at any given bloodflow rate; therefore, it may affect the balance of renal oxygen supply vs. demand and hence Epo production. The purpose of this study was to determine whether Epo production is modulated by the renin-angiotensin system (RAS). Twenty normal subjects on a controlled sodium and protein diet had glomerular filtration rate (GFR) and renal plasma flow (RPF)assessed by standard methods of inulin and para aminohippurate clearance, respectively, at baseline, hourly after the administration of losartan (25 mg) and after each 30 minute period of the infusion of Ang II at doses of 0.5, 1.5 and 2.5 ng/kg/minute. The baseline GFR was 115+/-4.0 ml/minute/1.73 m2, RPF 650+/-29 ml/minute/1.73 m2 and Epo 12.4+/ 0.8 U/L. In spite of a marked increase in filtration fraction (FF) with Ang II, no changes in serum Epo levels were observed at two hours(11.7+/-1.3 U/L, p=n.s. compared with baseline). After the administration of losartan, there was a variable effect on FF, but a strong correlation of the change in serum Epo concentration and the change in FF(r=0.648, p=0.002), suggesting that the RAS may modulate Epo production. PMID- 11881133 TI - [Specificity of host-parasite relations between arthropods and terrestrial vertebrates]. AB - Specificity of partners in host-parasite system is one of its main characteristics. Unfortunately this term has different senses in scientific literature. In everyday practice one judges an extent of host specificity of a parasite mainly by indices of its occurrence and abundance on different host species. An occurrence of parasites in nature reflects general result of complex eco-physiological interrelationships between partners in hostparasite system. Specificity of parasites in a choice of hosts may depend on a belonging of the latter to certain taxa (phylogenetic specificity), or on biotic and abiotic factors (ecological specificity). In arthropods, the phylogentic specificity and coevolution are characteristic to a greater extent for permanent hosts (lice, Mallophaga, cheyletoid and feather mites). A coevolutionaryphylogenesis is disturbed by transfers of parasites onto new hosts, by different rates of speciation in filial lines or by an extinction of several parasite taxa. In temporary parasites different forms of ecological specificity are prevalent. A host specificity is expressed to the lesser extent in mosquitoes, horseflies and in other blood-sucking Diptera. In temporary parasites with a long-term feeding (ticks) coevolutionary sequences are relatively rare, because this parasites had to adapt not only to a life on host, but also to a lesser stable environment. In some nest-burrow bloodsuckers (fleas, gamasid mites and argasid ticks) the ecological specificity is shown no by their relations with certain host species, but by an associations with habitats occupied by hosts (burrow, nests, caves). In relation with a high dynamics of host-parasite system, a specificity of its partners is comparative and it is kept up only under specific ecological conditions. PMID- 11881134 TI - [Histopathologic changes of bird skin in feeding places of ticks of the genus Ixodes (Acari: Ixodidae)]. AB - Histopathological changes of skin of Passeriformes birds in the places of attachment and feeding of the Ixodes persulcatus and I. lividus were investigated. The inflammation of bird skin is analogous to one of mammalian skin. The mouthparts of tick are surrounded by the fibrin cone and collagen capsula. They are formed by host. Ticks of these species do not form the cement. A proliferation stage of skin inflammation in birds is expressed. The thick collagen capsula develops in a result of proliferation. The feeding cavity is formed by the process of sucking a blood, inflammate cell infiltrate and products of tissue lysis. Neutrophils and eosinophils predominate in the inflammate cell infiltrate. PMID- 11881135 TI - [New species and sympatric relations of the chigger mite species group Talmiensis (Trombiculidae, Neotrombicula)]. AB - A revision of chigger mites species being closely related to Neotrombicula talmiensis (Schluger, 1955) has been performed. 2 new species are described: N. pontica sp. n. from Krasnodar Territory (Western Caucasus) and N. sympatrica sp. n. from Krasnodar Territory, Daghestan, Tuva, Armenia, Kirghizia and Turkey (Rize Province). N. pontica sp. n. is closely related to N. carpathica Schluger et Vysotzkaya, 1970 and differs from this species by the larger number of idiosomal setae (NDV = = 75-99 against 63-77), shorter legs (Ip = 782-847 against 844-920, TaIII = 67-74 against 70-80), lesser m-t (0.180 against 0.192), slightly lesser scutum and slightly longer setae. N. sympatrica sp. n. is closely related to N. carpathica and differs from this species by the longer scutal and idiosomal setae (PL = 67-78 against 57-69, H = 65-75 against 57-68, Dmin = 43-52 against 39-48, Dmax = 61-70 against 54-64), longer legs (Ip = 892-973 against 844-920, TaIII = = 77-86 against 70-80) and lesser m-t (0.168 against 0.192). N. carpathica is reported for the first time from Northern Caucasus (Karachai-Cherkess Republic, Kabardino-Balkaria, North Ossetia); N. talmiensis is reported for the first time from Khakasia. Data on joint occurrence of 3 species are reported. The 3 types of sympatric pairs of the species have been found in the Western Caucasus: 1) N. pontica sp. n. and N. sympatrica sp. n., 2) N. sympatrica sp. n., and N. carpathica, 3) N. pontica sp. n., and N. carpathica. Differences between species in that localities, where the joint occurrence was recorded, have been analysed. Sympatric relations between N. sympatrica sp. n. and N. carpathica are characterized by increased value of some general diagnostic characters of these species, such as the length of idiosomal setae and length of legs. Different characters play main distinguishing role in different sympatric localities. Besides that, some local features appear in certain species of sympatric pairs (narrow scutum in 3 samples of N. carpathica, numerous idiosomal setae in 1 sample of N. sympatrica), that differs the populations of the sympatric pair, and also discriminates such aberrant group from other material of the same species. Functions produced by the method of discriminant analysis are proposed for the purpose of diagnostics in talmiensis group. Tests for assumptions of discriminant analysis, such as normality and homogeneity of variances, has been performed beforehand. Verifications of the functions on test samples demonstrated the good quality of its working. Key to species using discriminant functions has been constructed. PMID- 11881136 TI - [New species of the Rhinonyssid mites (Gamasina: Rhinonyssidae) from birds of Russia and neighboring countries]. AB - Four new species of the nasal mite family Rhinonyssidae collected in different regions of the former USSR are described: Neonyssus (Otocorinyssus) alaudae sp. n. from Alauda arvensis L. (Alaudidae, Passeriformes) from Turkmenistan; Rhinonyssus clangulae sp. n. from Clangula hyemalis (L.) (Anatidae, Anseriformes) from Yakutia; R. marilae sp. n. from Aythya marilae L. (Anatidae, Anseriformes) from the Russian Far East; Locustellonyssus sibiricus sp. n. from Locustella certhiola (Pall.) (Sylviidae, Passeriformes) from Siberia. PMID- 11881137 TI - [On the development of body-wall musculature Diplostomum chromatophorum metacercariae (Trematoda: Diplostomidae)]. AB - The body-wall musculature of invasive cercariae and metacercariae of Diplostomum chromatophorum at different intervals after the penetration into the experimental intermediate host Cyprinus carpio (1, 3, 5, 6, 7, 10, 12, 20, 34, 40 days) has been investigated with the help of TEM technique. During the first 10 days after the invasion (in conditions of our experiment), the cercarial subtegumental muscle fibres degenerate. These muscles are replaced by newly formed ones. Mass differentiation of myoblasts beneath the tegument was observed in 7-10-day-old metacercariae. Obtained data indicate the metamorphosis of body-wall musculature during the morphogenesis in Diplostomum chromatophorum metacercariae. PMID- 11881138 TI - [Seasonal variations of the localization of the Bunodera luciopercae marites (Trematoda: Bunoderidae) in three species of perch fishes]. AB - Distribution of Bunodera luciopercae marites in the intestine of the fuff, sande and perch were studied in the Rybinsk Reservoir. For the fuff and perch fry, except three summer months, the more proportion of helminths was observed in the posterior region of the intestine. Because of a continuous passage of B. luciopercae in predatory fishes (sanders and adult perches), more proportion of trematodes was always observed in anterior region of the intestine. In all groups of hosts, 20-30% of total parasite number was registered in the middle intestine. PMID- 11881139 TI - [Dactylogyrids (Monogenea: Dactylogyridea) with unusual number of the anchors, their origin and phylogenetic significance. Original data]. AB - The haptors Dactylogyrus spp., Anacanthorus sp., Trianchoratus sp. and Schilbetrematoides pseudodactylogyrus are investigated. On the base of the morphology, transfer of the domus, etc. a homology of the hooks in dactylogyrids (s. s.) and the pin-like structures (4 "A") in the haptor of Anacanthorinae and Dactylogyridae sensu Bychowsky et Nagibina, 1978 is demonstrated. The vestiges of the anchors in the haptor of dactylogyrids (s. l.), according to morphological data, correspond to the point of anchor formed during the ontogenesis. In S. pseudodactylogyrus the pins are redescribed as typical vestiges of the anchors. PMID- 11881140 TI - [Aspidogasters--the parasites of unionids in the Zhitomir Polesye]. AB - The paper provides the data on the invasion of unionids in the Zhitomir Polesye by parasitic worms Aspidogaster conchicola. The aspidogasters have been found in 9 species of molluscs (Unio--5, Colletopterum--1, Pseudonodonta--1, Batavusiana- 2). An extensity and intensity of invasion of unionids by A. conchicola are unequal in different host species. The highest values of them were recorded for Unio conus borysthenicus (87% and 12 specimens), U. tumidus falcatulus (75%; 5 specimens), and Batavusiana nana (59.8%; 20 specimens). The invasion of molluscs by aspidogastrids from the stagnant waters are higher, than from the fluvial waters. The aspidogasters were found in the pericardium and kidneys of unionids. More often the parasites were met in the pericardium exclusively, rarely in the pericardium and kidneys simultaneously, and most rarely in the kidneys only. Within the pericardium, A. conchicola prefer the furthest sites of the contractile ventricle of the molluscan heart. PMID- 11881141 TI - Here's kitty, kitty. PMID- 11881142 TI - The next big thing in health benefits: consumer choice. AB - Consumers are the only ones who can affect all decision points that drive health care cost and quality. As a result, consumers' health and financial security depend on their taking more responsibility for their health care decisions and having the tools and information needed to do so successfully. This article explains the five key decision points that drive health care cost and quality, how technology aids marketplace innovations, and how employers can help advance consumer choice in order to push the health care system to deliver better care and keep inflation in check. PMID- 11881144 TI - Complete preemption--state law alternative enforcement mechanism--medical malpractice. Danca v. Private Health Care Systems, Inc. Thompson v. Gencare Health Systems, Inc. PMID- 11881143 TI - HIPAA's privacy rule: what is it, and how does it affect you? AB - This article discusses the implications for stakeholders of the privacy rule under the Health Insurance Portability and Accountability Act of 1996, on which the U.S. Department of Health and Human Services first released guidance on July 6, 2001. Although guidance will continue to evolve, the authors urge organizations to initiate the implementation of policies to ensure compliance by the actual effective date of 2003. (Small health plans have until 2004 to comply.) PMID- 11881146 TI - Preemption--HMO doctors' failure to disclose financial incentives- misrepresentation and fraud. Shea v. Esensten. PMID- 11881145 TI - Preemption--federal jurisdiction--medical malpractice. Bauman v. U.S. Healthcare, Inc. PMID- 11881147 TI - [Cytokines--a general system of homeostatic regulation of cell functions]. AB - The paper reviews biological characteristics of cytokines, which are short living molecules exerting pleiotropic and redundant effects on a variety of target cell types, influencing cell activation and differentiation. Significance of cytokines for patho- and immunogenesis of infections and other diseases is analysed, in addition to further perspectives of cytokine employment for prophylaxis and treatment of various diseases. PMID- 11881148 TI - [Monokine-producing activity of human monocyte-like cell line U937 induced by Yersinia pestis EV antigens]. AB - The data on a study of the monokine-producing ability of human monocyte-like cell line U937 are presented. Antigens of Yersinia pestis EV (lipopolysaccharide and fraction 1A) induce monokine production by cell line U937. The obtained monokines essentially enhance neutrophil killer and chemotactic activities, stimulate FcR expression, increase the number of lysosomes, and the lability of lysosomal membranes in neutrophils. F1A significantly suppresses LPS in respect to the ability to induce monokine production, which stimulate neutrophil functional activity. PMID- 11881149 TI - [Information EHF-interactions in a system of live objects (human platelets)]. AB - The molecular informational interaction has been first detected in a system that involves human platelets, exposed to electromagnetic EHF-fluctuations at frequencies of molecular spectra of radiation and absorption of nitric oxide (150.176-150.644 HHz), and native platelets. It has been established that the incubation of a native platelet rich plasma with a similar plasma, exposed to a 5 minute effect of electromagnetic EHF-fluctuations at frequencies of molecular spectra of radiation and absorption of nitric oxide at a mode of peak and frequent modulation of a signal under in vitro conditions, causes a significant (P < 0.05) inhibition of platelet functional activity in the native plasma, in comparison with control. This was displayed by a decreased platelet activation and falling platelet aggregation ability. Some possible mechanisms of interaction are suggested to explain the described effect. PMID- 11881150 TI - [Monoclonal antibodies against protein Daxx and its localization in nuclear domains 10]. AB - Nuclear domains 10 (ND10) were first detected occasionally using antibodies to an antigen of unknown nature (Ascoli, Maul, 1991). Further on it was shown that ND10 were sites of locality of the number of proteins (PML, Sp 100, pRB) (Sterndorf et al., 1992; Kamitani et al., 1998), the majority of which are modified with ubiquitin-like small proteins-modifiers (SUMO) (Ishov, Maul, 1996). In addition, it was shown that ND10 were sites of primary localization, transcription and replication of some DNA-viruses (SV40, virus of simple herpes 1, adenovirus 5) (Ishov, Maul, 1996; Ishov et al., 1997). Except for SV40, these viruses produce proteins able to modify ND10, or leading to degradation of ND10-associated proteins (Maul et al., 1993; Maul, Everett, 1994). This degradation is accompanied with protein desumofication and, later, with hydrolysis on the ubiquitin-proteosomal way (Everett et al., 1998, 1999). Cell incubation with interferon leads to augmentation of the number and dimension of ND10 owing to increased expression of Sp100 and PML (Lavau et al., 1995; Grotzinger et al., 1996). In all, these data make it possible to put forward a hypothesis that ND10 may represent a peculiar cell storage ("depot") of proteins regulated according to the "accumulation-drop" principle (Ishov et al., 1997; Maul, 1998). However, this hypothesis requires further factual grounds. PMID- 11881151 TI - [Involvement of SH-groups in the regulation of adenylate cyclase signal system by isoproterenol and serotonin in cultured murine fibroblasts L (subline LSM)]. AB - The adenylyl cyclase system (ACS) plays a key role in transduction of a hormonal signal into eukaryotic cells. The functional activity of the system depends on SH groups of proteins involved in the ACS: receptor, G-protein, and enzyme adenylyl cyclase (AC). We studied the influence of thiols and SH-blockers on the regulation of AC activity by nonhormonal (NaF and Gpp[NH]p) and hormonal (biogenic amines isoproterenol and serotonin) agents in homogenates of cultured murine fibroblasts of line L (subline LSM). In the presence of thiols 2 mercaptoethanol (5 mM) and dithiothreitol (1 mM) the basal AC activity somewhat increased, whereas the stimulating effects of NaF, Gpp[NH]p, and hormones decreased. No potentiating action of Gpp[NH]p on hormonal effect in this case was found. The SH-blockers 25 mkM p-chloromercuribenzoic acid (CMBA) and 0.2 mM N ethylmaleimide significantly inhibited both the basal AC activity and that stimulated by different agents. Thiols partially restored CMBA inhibited AC activity (in the case of N-ethylmaleimide restoring effects of thiols were insignificant). This, the ACS of murine fibroblasts of subline LSM is SH sensitive. The forms of SH-groups in proteins involved in the ACS determine their functional activities and a possibility of transduction of the hormonal signal on the effector systems. PMID- 11881152 TI - [The role of Src-kinase in the regulation of endocytosis of EGF-receptor complexes. I. Dynamics of EGF internalization, recycling, sorting, and degradation during inhibition of Src-kinase activity]. AB - In the present work, the role of Src-kinase in regulation of different stages of EGF-receptor endocytosis was studied. We used murine fibroblasts with knockout of Src gene and CGP77675, and the inhibitor of Src-family kinases. The absence of Src protein in the cells did not lead to any changes in the rates of 125I-EGF internalization or recycling and caused only slight decrease in the rate of its degradation. At the same time, treatment of the wild type cells with the inhibitor resulted in a small decrease in internalization rate and an increase in recycling. The influence of the inhibitor on 125I-EGF degradation was also more pronounced. But even in this case, the effects were no more than 30% of control values. CGP77675 extended the same effect upon cells of HER14 and HC11 lines. Subcellular fractionation of these cells in Percoll gradient has also demonstrated a slight inhibition of 125I-EGF sorting from early to late endosomes. The more pronounced effect of the Src-family kinase inhibitor on the EGF endocytosis, compared to that of the absence of a single Src protein, suggests a compensating mechanism of the Src-family kinases. A conclusion is made that in spite of a slight influence on practically all stages of intracellular pathway of EGF-receptor complexes, Src-kinases are obviously not the key regulators of their endocytosis. PMID- 11881153 TI - [The role of Src-kinase in the regulation of endocytosis of EGF-receptor complexes. Distribution of clathrin after stimulation of EGR endocytosis in various cell lines during inhibition of Src-kinase activity]. AB - A distribution of EGF receptor and clathrin during EGF endocytosis in A431, HER14, WT and PURO cell lines was studied by indirect immunofluorescence. Though the initial distribution of EGF-receptors on A431 and HER14 cells was somewhat different, the late stages of endocytosis proceeded equally and were marked by formation of bright spots in the juxtanuclear region characteristic of the late endosomes. The Src-family kinase inhibitor CGP77675 had no influence on the dynamics of receptor endocytosis at the immunofluorescent level in both cell lines. Stimulation of EGF-receptor endocytosis in A431 cells did not also result in any redistribution of clathrin in the areas where the majority of EGF receptors are localized, i.e. in the lateral plasma membrane both in the control cells and under CGP77675 treatment. Clathrin in A431, WT and PURO cells demonstrated even a punctuated pattern throughout the cytoplasm with some accumulation in the juxtanuclear region. This distribution depended neither on the absence or presence of Src activity nor on EGF addition. The data obtained indicate that 1) EGF-receptors do not serve as the initiation sites during clathrin coated pit assembly; 2) Src-kinase activation does not result in significant clathrin redistribution in the plasma membrane, and its influence on EGF endocytosis can be considered as a secondary effect. PMID- 11881154 TI - [H2O2-induced activation of transcription factors STAT1 and STAT3: the role of EGF receptor and tyrosine kinase JAK2]. AB - Reactive oxygen species initiate multiple signal transduction pathways including tyrosine kinase signaling. Here, we demonstrate tyrosine phosphorylation of EGF receptor, STAT3, and, to a lesser extent, STAT1 upon H2O2 treatment of HER14 cells (NIH3T3 fibroblasts transfected with full-length EGF receptor). Maximum phosphorylation levels were observed in 5 min of stimulation at 1-2 mM H2O2. It has been shown that the intrinsic EGF-receptor tyrosine kinase is responsible for the receptor phosphorylation upon H2O2 stimulation. STAT3 and STAT1 activation in HER14 cells was demonstrated to depend on EGF receptor kinase activity, rather than JAK2 activity, while in both K721A and CD126 cells (NIH3T3 transfected with kinase-dead EGF receptor, and EGF receptor lacking major autophosphorylation sites, respectively) STAT1 and STAT3 tyrosine phosphorylation requires JAK2 kinase activity. Furthermore, STAT3 is constitutively phosphorylated in K721A and CD126 cells, and STAT1 H2O2-stimulated activation in these cells is much more prominent than in HER14. In all the cell lines used, Src-kinase activity was demonstrated to be unnecessary for ROS-initiated phosphorylation of STATs. Herein, we postulate that EGF receptor plays a role in H2O2-induced STAT activation in HER14 cells. Our data also prompted a hypothesis of constitutive inhibition of JAK2-dependent STAT activation in this cell line. PMID- 11881155 TI - [The role of Mac-1 and ICAM-1 molecules in adhesion of cells on fibronogen and its degradation products]. AB - Monocytic cell adhesion to immobilized fibrinogen and fibrinogen degradation products, and involvement of integrins Mac-1 and immunoglobulin-like ICAM-1 adhesion molecules in these processes were investigated. Fibrinogen cleavage with plasmin down-regulated adhesion of cells with predominant Mac-1 expression; in contrast, the attachment of ICAM-1-expressing was up-regulated. By means of function-blocking anti-Mac-1 and anti-ICAM-1 antibodies, and immobilization of known fibrinogen degradation products, it was shown that Mac-1 molecules mediated cell adhesion predominantly to fibrinogen, and its early degradation products, fragments X and Y, while ICAM-1 participated in cell attachment to X- and Y fragments, rather than to intact fibrinogen or late degradation products, fragments D and E. PMID- 11881156 TI - [Radioadaptive enhancement of the repair of UV-induced postreplication gaps in Escherichia coli DNA]. AB - Postreplication DNA repair (PRR) in UV-irradiated Escherichia coli WP2 uvrA (tryptophan-dependent strain) and K12 AB1886 uvrA6 pre-irradiated by gamma-rays in low doses (radioadaptation, the first stress effect) has been investigated. PRR was found to be more effective after incubation in the growth medium (for 45 60 min) than in non-radioadapted cells: the repair of postreplication gaps increased by 6-15%. If cells of WP2 uvrA strain were incubated after UV irradiation in media lacking tryptophan or casamin acids (the second stress effect), PRR was seen to increase as early as within 15 min of incubation and it is more effective than at the first stress. After a 30-60 min incubation the double stress effect leads to an increase in postreplication gap repair by 23 45%. In this case almost all the gaps prove to be repaired. The second stress alone exerts no influence on PPR efficiency. It is supposed that a preliminary radioadaptation may stimulate synthesis of a protein (proteins) of the SOS response (presumably DNA polymerase V). The second stress effect apparently induces synthesis of an unknown factor (or depreesses synthesis of a MmrA-like protein), and this in cooperation with a protein newly synthesized during radioadaptation significantly increases the efficiency of PPR. PMID- 11881157 TI - [Protein synthesis in kidneys of merino sheep in postnatal ontogenesis]. AB - By evaluating parameters of activity of the nucleolar organizers (AgNORs), patterns of protein synthesis in cells of the right and left kidneys of merino sheep were examined in dependence of their age, sex, and the organ position. Depending on these parameters, the number of AgNORs in the kidney of merino sheep varies from 2 to 7. The total area of AgNORs in cells was within 0.50 +/- 0.03 2.22 +/- 0.05 mcm2. In different structure of nephrons, regions of nuclear organizers differ in both the quantity and total area. The obtained data testify that in kidney of 3 month old males and females of merino sheep the synthesis of 18S- and 28S-classes of ribosomal RNA and, accordingly, proteins occurs most actively. PMID- 11881158 TI - Avoidant personality disorder: current status and future directions. AB - Avoidant Personality Disorder (APD) is the topic of a growing body of research literature. In this article, we review empirical studies of APD with the goals of identifying the themes that underlie this work and pointing to new directions for future research. In particular, we recommend that future studies evaluate several unique factors postulated by personality theorists to be central to this condition. PMID- 11881159 TI - Treatment outcome for chronic PTSD among female assault victims with borderline personality characteristics: a preliminary examination. AB - The present study examined the effect of the presence of borderline personality disorder characteristics (BPC) on patients' responses to cognitive behavioral treatment for chronic PTSD. Seventy-two female victims of sexual and non-sexual assault were randomly assigned to one of four treatment conditions: prolonged exposure (PE), stress inoculation training (SIT), combined treatment (PE/SIT), and wait list control (WL). Treatment consisted of nine bi-weekly individual sessions. Seventeen percent of the patients met full (10%) or partial criteria (7%) for borderline personality disorder. A greater number of patients with BPC reported sexual assault in childhood compared to those without the symptomatology. Patients with BPC also reported more pre-treatment anger. In general, those with BPC benefited significantly from treatment, although at post treatment, they were less likely to achieve good end-state functioning than those without such symptomatology. The relationship between BPC and treatment response will be discussed. PMID- 11881160 TI - Authority conflicts and personality disorders. AB - We investigated personality disorders (PDs) of young men with chronic authority conflicts using the Millon Clinical Multiaxial Inventory-I (MCMI-I; Millon, 1983). PDs occurred considerably more often in the young men with chronic authority conflicts (62%) compared with a clinical control group of young men with acute authority conflicts (39%). The prevalence of PDs in a normal control group was considerably lower (11%). The passive-aggressive PD is over-represented among young men with chronic authority conflicts; there is hardly any antisocial PD. Millon's conceptualization of these disorders could have influenced these results. The high prevalence of passive-aggressive PDs complicates a good psychotherapeutic outcome in young men with chronic authority conflicts. Our results underscore the clinical utility of the controversial PAPD diagnosis. PMID- 11881161 TI - Histrionic personality disorder and antisocial personality disorder: sex differentiated manifestations of psychopathy? AB - Little is known about the etiology of histrionic personality disorder (HPD) or its relation to other personality disorders. In this study, we examined whether HPD is etiologically related to psychopathy and more specifically whether HPD and antisocial personality disorder (ASPD) are sex-typed alternative manifestations of psychopathy. In addition, based on Newman's (1987) response modulation hypothesis of psychopathy, we examined the associations between psychopathic, HPD, and ASPD features and performance on laboratory measures of passive avoidance errors and interference effects. Seventy-five live theater actors completed self-report questionnaires and two laboratory measures of response modulation, and peers completed questionnaires concerning the participants' personality disorder features. The results provided weak and inconsistent support for the hypotheses that HPD is a female-typed variant of psychopathy and that ASPD is a male-typed variant of psychopathy. Contrary to previous findings, scores on response modulation tasks were not significantly related to psychopathy, or to either HPD or ASPD. The limitations of this study and possibilities for future research in this area are outlined. PMID- 11881163 TI - Determinants of diagnostic prototypicality judgments of the personality disorders. AB - We evaluated the impact of three factors that have been suggested to be important determinants of the diagnostic process pertaining to personality disorders: (a) the number of features representative of a personality disorder category; (b) the extent to which those features are typical of the category, and (c) the dominance or proportion of category features to the total number of features. Thirty-two clinical psychologists evaluated 12 profiles of hypothetical patients in which these factors were varied factorially. The results revealed strong effects of typicality and dominance, but no effects for number. Implications for the assessment and classification of personality disorders are discussed. PMID- 11881162 TI - The association between axis I and II psychiatric symptoms and high-risk sexual behavior during adolescence. AB - This study was conducted to investigate the association between psychiatric disorders and high-risk sexual behavior among adolescent primary care patients. Interviews assessing anxiety, conduct, depressive, eating, substance use, and personality disorders (PDs), as well as histories of sexual behavior were administered to 119 male and 284 female adolescent primary care patients. Results indicated that, after co-occurring psychiatric disorders were controlled statistically, adolescents with elevated PD symptom levels were more likely than adolescents without elevated PD symptom levels to report a high number of sexual partners during the past year and during their lifetime. Adolescents with a history of conduct disorder were more likely than adolescents without such a history to report a high number of lifetime unsafe sexual partners. Elevated antisocial, dependent, and paranoid PD symptom levels were associated with high risk sexual behavior after co-occurring psychiatric disorders were controlled. Further, certain specific antisocial, borderline, dependent, histrionic, narcissistic, obsessive-compulsive, paranoid, and schizotypal PD symptoms were independently associated with high-risk sexual behavior after co-occurring psychiatric disorders and overall PD symptom levels were controlled. The association between overall PD symptom levels and the number of sexual partners was significantly stronger among the females than among the males in the sample. Increased recognition and treatment of PDs, coupled with increased recognition of high-risk sexual behavior may facilitate the prevention of sexually transmitted diseases and teenage pregnancy among adolescents. PMID- 11881164 TI - Smallpox visits the White House. PMID- 11881165 TI - Tuberculosis--elimination in the third millennium? PMID- 11881166 TI - Steroid therapy in chronic obstructive pulmonary disease. AB - COPD is a prevalent disease, with an increasing attributable mortality. Because inflammation plays a significant role in the pathogenesis of this disease, the use of anti-inflammatory therapies would appear indicated; hence the widespread use of corticosteroids in COPD. Although the majority of patients with stable COPD do not benefit from systemic steroids, there is good evidence supporting the use of short courses of systemic steroids for COPD exacerbations. With respect to inhaled corticosteroids, the studies are conflicting. Those patients with an asthmatic component to their disease, or with a positive bronchodilator test, appear to benefit most from inhaled steroids. Those with irreversible disease do not benefit from short-term inhaled steroids. Long-term inhaled corticosteroids, though not having a significant effect on the rate of decline in spirometric indices, do appear to decrease the number of exacerbations and the rate of decline in health status, reduce respiratory symptoms, decrease use of health care services, and improve airway reactivity. These effects appear more marked in patients with moderate-to-severe disease. Because very few therapies offer significant benefits to patients with COPD, and until a test is developed that will distinguish between potential steroid responders from non-responders, it is worthwhile giving all patients with COPD a trial (3-6 months) of inhaled corticosteroids to determine whether they are responsive. PMID- 11881167 TI - Obstructive sleep apnea: a brief overview for the primary care physician. PMID- 11881168 TI - End of life issues in the critically ill. PMID- 11881169 TI - Retrotracheal parathyroid adenoma. PMID- 11881170 TI - Update on treatment for congestive heart failure. PMID- 11881172 TI - Quick reference guide for asthma diagnosis and treatment. PMID- 11881171 TI - Health status, access to care, and health risk behaviors among urban Rhode Islanders, 2000. PMID- 11881173 TI - Point of view: Marfan syndrome: be aware of life-threatening complications. PMID- 11881174 TI - Neighborhood Health Plan of Rhode Island. PMID- 11881175 TI - Bioterrorism and physicians. PMID- 11881176 TI - No, hatred is not what we need. PMID- 11881177 TI - Women's use of resources in leaving abusive relationships: a naturalistic inquiry. AB - Studies have considered the experiences of women in terminating abusive relationships. However, the personal strengths and internal resources women use to assist them in terminating have not been studied. The purpose of this naturalistic inquiry was to explore what personal strengths and inner resources contributed to (1) the actual making of women's decision to terminate the abusive relationship, and (2) their ability to successfully follow through with this decision. Themes of turning point, realization, reframing, agency, and self efficacy emerged from the study. The results suggest a process, although the steps were not sequential and were repeated numerous times before the women were finally successful in leaving. Based on results of this study, implications for intervention include the promoting of cognitive approaches such as reframing and self-talk to facilitate the process of leaving. PMID- 11881178 TI - Sex offender treatment: therapists' experiences and perceptions. AB - The literature portrays work with sex offenders as mentally, physically, and emotionally draining. However, the therapists working in the outpatient Sexual Abuse Treatment (SAT) program at a regional mental health center, while not denying the challenges of this work, describe their experiences as generally positive and rewarding. Because these therapists' experiences seemed to contrast with the literature, a qualitative descriptive study was conducted to examine the SAT therapists' experiences and perceptions. Seventeen SAT therapists were individually interviewed. Interviews were audio taped, transcribed verbatim, and analyzed using constant comparative analysis. The therapists in this study indicated that although sexual offender treatment is a stressful area to work in, it offers many rewards and is viewed as "a challenge and privilege." Exploring the positive as well as the negative impacts of working with sex offenders may help identify additional coping strategies or mediating factors for this challenging work. PMID- 11881179 TI - Special observation in the care of psychiatric inpatients: a literature review. AB - This paper summarizes the results of a search of electronic databases for papers on special observation (SO). Published studies to date about SO are entirely descriptive. No evaluative research appears to have taken place, leaving the procedure based on clinical pragmatism and tradition. Something between 3%-20% of admissions receive some form of SO during their stay and the rate of usage varies widely between wards. SO is used as a method of controlling and containing the most disturbed patients who are considered to be imminently at risk of harming themselves or others. Such patients tend to be younger and suffering from acute psychosis or depression. Which professional staff have the authority to initiate and terminate SO varies from place to place, as does its duration. The financial costs have been crudely assessed and are reported to be very high, perhaps up to 20% of the nursing budget for a hospital. Further variation exists on who is allowed to carry out SO. Nurses frequently make unofficial modifications to the procedure based upon their own individual judgments and assessments, and policies vary widely among hospitals. There is little agreement between authorities on what nurses should do during SO, although there is some evidence that it can, under certain circumstances, be therapeutic. However there is also evidence that nurses find SO stressful and patients dislike it. PMID- 11881180 TI - The practices of expert psychiatric nurses: accompanying the patient to a calmer personal space. AB - The focus of the care of potentially aggressive psychiatric patients has been on the use of seclusion and restraints. Recent concerns, however, about the potential for patient injury have made it imperative that nurses use alternative methods to calm patients who are escalating. Little is known about how expert nurses de-escalate the escalating patient. The purpose of this interpretive phenomenological study was to uncover and describe the knowledge embedded in the stories of psychiatric nurses who are skilled in the practices of de-escalating an escalating patient. Twenty registered nurses were interviewed using an unstructured format. The analysis of the data revealed that these nurses were skilled at noticing the patient, reading the situation and the patient, knowing where the patient was on the continuum, understanding the meaning of the behavior, knowing what the patient needed, connecting with the patient, and matching the intervention with the patient's needs. PMID- 11881181 TI - The use of aberrant medical humor by psychiatric unit staff. AB - The purpose of this study was to develop a grounded theory about the joking behaviors of psychiatric unit staff. Fifty nine staff members (Male = 23, Female = 36), ages 28-62, who worked in an urban, public facility were observed in unit meetings over a two year period. Data were analyzed using the constant comparative method. Participants used aberrant humor to deal with the basic social process of facing a series of ultimately unresolvable problems. The factors involved included the organizational context of the working environment, contact with a stigmatized patient group, and stressful staff-patient interactions which led to the erosion of a sense of professional competence and the resulting demoralization, resentment, and vengeful counteraction. Two major categories of joking emerged: whimsical and sarcastic humor. Six subcategories of humor also emerged: incongruous, bravado, self-mocking, discounting, malicious and gallows humor. Staff engaged in more sarcastic than whimsical humor. Findings support the need for professionals to recognize the potentially detrimental effects of joking behaviors on their treatment effectiveness. PMID- 11881182 TI - The relationship between basic need satisfaction and emotional eating. AB - Eating in response to emotions may lead to the consumption of excessive calories which typically leads to weight gain. This study examined the relationship between basic need satisfaction as identified by Maslow's hierarchy and emotional eating. According to Modeling and Role-Modeling theory, when lack of basic need satisfaction functions as a stressor, individuals may be more likely to engage in emotional eating as a substitute for fulfilling their needs in order to maintain homeostasis. The Basic Need Satisfaction Inventory (BNSI) had a strong, negative correlation (r = -.49; p < .001) to the Emotional Eating Scale (EES) indicating that the lower the level of basic need satisfaction, the more likely one engaged in emotional eating. In predicting EES score, 27.7% of the variance was explained by the self-esteem subscale of BNSI. This study supports looking at underlying issues contributing to weight gain in order to develop effective interventions for weight management. PMID- 11881183 TI - Retirement planning, use of time, and psychosocial adjustment. AB - This was a descriptive study to examine the relationship between retirement planning, use of time, and the psychosocial adjustment to retirement. A 72-item questionnaire was mailed to all retirees (N = 1565) from an international company residing in seven southeastern states with a response of 764 (48.82%). Significant differences were found when comparing what retirees indicated they had done to prepare for retirement, and what they believed after retirement was important for people to do to prepare. Following retirement greater emphasis was placed not only on financial factors, but also on psychosocial adjustment and use of time. Findings suggest that retirement as a psychosocial transition needs to be an integral component of nursing assessments and interventions for retirees and their families. PMID- 11881184 TI - Here, kitty, kitty! PMID- 11881186 TI - In praise of folic acid. PMID- 11881185 TI - On their own two feet. PMID- 11881187 TI - [Comparative yearly growth rate of children with mucoviscidosis treated and not treated with ciprofloxacin:clinicomorphological comparisons]. AB - The results of the prospective and comparative investigation of the linear growth of children at the age of 4 to 16 years with mucoviscidosis treated with ciprofloxacin in combination with a cephalosporin or an aminoglycoside in the main group and a cephalosporin or an aminoglycoside alone in the control group are presented. The children were observed for 3 and 5 years. It was shown that in spite of the treatment term with ciprofloxacin the yearly growth rate in the children in the main and control groups did not significantly differ. The morphological investigation did not reveal any injury of the armicular cartilage and growth zone. The hyperplastic reaction in the tegmental cartilage was states and considered as a physiological one in response to the presence of ciprofloxacin and therefore reversible. No chondrotoxicity of the fluoroquinolones and ciprofloxacin, particularly in the children, is explained. PMID- 11881188 TI - [Evaluation of amixine reactivity and efficacy for prophylaxis of acute respiratory tract infections]. AB - Amixine reactivity and tolerability were evaluated in controlled trial at the risk group of medical personal at the period of flu and respiratory viral infection season. Drugs safety was estimated according to anamnesis, direct observation and hemogram. High efficacy of the drug for the infections prophylaxis and treatment was demonstrated. The drug was well tolerated and had no side effects. Amixine unreactivity was proved. PMID- 11881189 TI - [Empiric antibacterial therapy of acute intestinal infections in children (manual for physicians)]. PMID- 11881190 TI - [Start treatment of community-acquired pneumonia in outpatients and in hospitalized patients]. PMID- 11881191 TI - [Natural immune system cells and perspectives to develop recombinant proteins with microbial activity]. PMID- 11881192 TI - [Cefoperazone (Medocef*) in the modern therapy of severe bacterial infections]. PMID- 11881193 TI - [Synthesis and antibacterial activity of the new 9-aminoacridine, 10,11-dihydro 5H-dibenzo[b,f]azepine, polyfluoro 5,6-dihydro-1,3,5-oxadiazine derivatives]. AB - Screening among 9-aminoacridine, 10,11-dihydro-5H-dibenz[b, f]azepine and polyfluoro 5,6-dihydro-1,3,5-oxadiazine derivatives allowed to isolate compounds with potential antibacterial activity. Schemes of the active compounds synthesis are given. The most important is the estimation of the oxydiazines activity against gram-positive microorganisms including methicillin-resistant staphylococci. Special attention is paid to the activity of iminodibenzyl derivatives against multiresistant gram-negative microorganisms. PMID- 11881194 TI - [Problems of introducing hospital diagnostic technologies into ambulatory polyclinc facilities]. PMID- 11881195 TI - [Errors in diagnosing and treating injuries from toxic agents subject to elimination]. PMID- 11881196 TI - [Frostbite in modern fighting pathology]. AB - In modern war 1-2% in the structure of fighting surgical pathology constitute the frostbites. In cold time of year their frequency increases up to 10%. Late admission of the injured to the stages of medical evacuation, their different and often limited diagnostic and therapeutic potentials influence on the outcomes of frostbites' treatment. The role of rational operative treatment with the use of surgical dermato-plastic methods rises. The analysis of periods, outcomes of treatment, restoration of working capacity and fitness for military service after the frostbites of different degrees and levels of injury permits to propose the military-and-medical classification of this trauma type. Slight, moderate and severe frostbites were selected. The places of final treatment of the injured with reference to the classical scheme of treatment- and evacuation support under conditions of the local armed conflict were determined. PMID- 11881197 TI - [Significance of computer tomography and diagnosis of gunshot craniocerebral injuries at the stage of specialized medical care]. PMID- 11881198 TI - [Use of serotonin adipinate for improving the contractile activity of the hypokinetic zone of the myocardium]. PMID- 11881199 TI - [Paroxetine (paxil): use in clinical practice]. PMID- 11881200 TI - [Medical aspects of creating a technical "telemedicine" program]. PMID- 11881201 TI - [Project "Achilleosis"-onychomycosis at the turn of the millenium]. PMID- 11881202 TI - [Epidemiologic effectivness of poludane in preventing influenza and other acute respiratory diseases in troops]. AB - The epidemiological effectiveness of interferon inductor "Poludanum" produced by "LENS Pharm" (Russia) for prevention of acute respiratory viral infections was tested. The drug was administered among the students of Military Medical Academy (101 students) according to scheme 1: in a nose, 3 drops in each nasal passage two times per week during 4 weeks, with 2 week interruption then the scheme was repeated. The scheme 2 included 3 drops in each nasal passage once a week during 10 weeks (95 students). The placebo group (96 students) received the distilled water. In the students receiving "Poludanum" according to scheme 1 the incidence of acute respiratory diseases (ARD) was significantly lower than in the control group (p = 0.058). The coefficient of effectiveness was 50%, i.e. the incidence has decreased in two times. The use of poludanum according to scheme 2 caused no decrease in ARD incidence. PMID- 11881203 TI - [Modern trends in creating biologically active materials for treating purulent wounds]. PMID- 11881204 TI - [Pharmacologic correction of physical ability to work under common and complicated conditions]. PMID- 11881205 TI - [My path in military medicine]. PMID- 11881206 TI - [Words about a teacher. (on the 80th anniversary of the birth of A. N. Senenko)]. PMID- 11881207 TI - [Problems and prospects of post-graduate training at military-medical cadres]. AB - The actual problems of advanced post-diploma training of RF AF medical cadres is analyzed in the article. The investigation of paramount directions of military and-medical education reform was conducted, the data on additional military-and medical education system were generalized. The present average annual output of all bases of additional education in RF AF system was subjected to analysis. The data on the students' training cost in the State institutions for physicians' advanced training (SIPAT) and the same civilian medical higher schools are presented. It is noted that the one student training prime cost in RF AF SIPAT is considerably lower. Present conceptions concerning the advanced system of military-and-medical cadres' training were analyzed. The main problems of RF AF SIPAT are presented; and until the problems are solved it will be impossible to optimize the training of medical service specialists. It is shown that under conditions RF AF, military- and medical reformation SIPAT can become the leading educational, scientific and medical center of post-graduated and additional professional education of military physicians. PMID- 11881208 TI - [Major General V. M Adamovich--founder of the Moscow Military Hospital]. PMID- 11881209 TI - [Omsk garrison hospital--225 years]. PMID- 11881210 TI - [Yaroslav military hospital--130 years]. PMID- 11881211 TI - [Genetic aspects of species structure of the compost worm Eisenia foetida (Sav.) (Oligochaeta, Lumbricidae)]. AB - Distribution of frequencies alleles of polymorphous loci of peroxidase (Pox), leucineaminopeptidase (Lap), phosphoglucomutase (Pgm) and octanoldehydrogenase (Odh) were studied by electrophoresis in polyacrylamide gel in 22 local samples of Esenia foetida in Russia (European part), Ukraine, Kazakhstan and Kirghizia. The samples form two spatial groups--"northern" and "southern", distinguished by set of alleles in every studied locus. The "northern" groups is formed by local populations of European Russia from Murmansk region on the north to Smolensk region on the south, and also by cultivated population of selection line "red California hybrid". The "southern" group is formed by local populations on the territory of Russia from middle Volga to the North Caucasus, Ukraine, Kazakhstan, Kirghizia, cultivated populations from Kirghizia and Portugal. High degree of genetic difference between samples and independence of alleles frequencies distribution from geographical location and habitat allows to consider almost all studied groups as separate populations. Statistical processing of Nei genetic distances (Nei, 1972) revealed reliable differences between averages of within- and intergroup distances. Besides, discrete differences between intervals of significance of genetic distances were revealed. The results indicate that on the studied territory E. foetida has hierarchical two level structure. The first level is formed by local populations differed by frequency of the same alleles. The second level is formed by local populations, united into spatial groups, that are qualitatively distinguished by the set of alleles in the same loci. PMID- 11881212 TI - [Decentralized self-regulation of the integrity of colonial organisms]. AB - The self-regulation of the integrity of colonial organism is accomplished without any central system organs. Abundance of similar structures, parallelism and synchrony of the processes form the basis for self-regulation, thus creating the independent mechanism of the internal coordination of the processes. Decentralized regulation of the integrity is much more economical then the centralized regulation, because it does not demand large volume of information, consequent increase in complexity of its processing and unavoidable problems with coordination. The principles of decentralized self-regulation, addressed in this paper, can be use not only in biology, but also in sociology, psychology, modeling, neuroformatics, etc. PMID- 11881213 TI - [Does population ecology have general laws?]. AB - There is a widespread opinion among ecologists that ecology lacks general laws. In this paper the author argues that this opinion is mistaken. Taking the case of population dynamics, the author points out that there are several very general law-like propositions that provide the theoretical basis for most population dynamics models that were developed to address specific issues. Some of these foundational principles, like the law of exponential growth, are logically very similar to certain law of physics (Newton's law of intertia, for example, is almost a direct analogue of exponential growth). The author discusses two other principles (population self-limitation and resource-consumer oscillations), as well as the more elementary postulates that underlie them. None of the "laws" that the author proposes for population ecology are new. Collectively ecologists have been using these general principles in guiding development of their models and experiments since the days of Lotka, Volterra, and Gause. PMID- 11881214 TI - Role of the skeleton in determination of the branching points in hydroid colonies. AB - Colonial hydroids of the suborder Thecaphora have rigid outer skeleton that possesses species-specific shape of the colony elements. Organisation of the elements within a colony shows strict spatial patterning. The points of branching (emergence of the new growing tip) within shoots are strictly determined and show positive correlation with the place of the most pronounced curvature of the elements skeleton. As it was shown earlier, the shoot growing tip, after emergence, performs its program of functioning independently from the colony condition. Several modifications of experiments with grafting of the growing tip together with rotation around its longitudinal axis were fulfilled. As the result of such grafting the normal orientation of the formed skeleton was altered. In overwhelming cases of further tip growth and shoot development the new growing tip emerged in correspondence with new orientation of the formed skeleton. The orientation of the hydranth of the maternal shoot element had no effect upon the orientation of the new tip emergence after grafting. It is supposed that the place of the new tip emergence is regulated hierarchically. At first level, the interaction of the soft tissues with the skeleton has priority. Anisotropy of mechanical tensions within cell layers due to interaction with the skeleton at the point of its most curvature serves as a primary stimulus in the chain of events leading to the initiation of the new growing tip. If this does not work the intrinsic tissue property (polarity) determines the place of the tip emergence. Such two-level mechanism of determination of the place of the new tip emergence in sympodial shoots stabilises spatial organisation of the entire shoot and decreases the probability of epigenetic mistakes. PMID- 11881215 TI - [Effect of variable phases of tide cycle on reproduction of Laomedea flexuosa (Hydroidea, Thecaphora]. AB - Colonial hydroid Laomedea flexuosa inhabits the narrow belt of low littoral zone in the White Sea. What is a reason of so limited habitat? The authors studied the time of planulae release and its behavior during free swimming stages and settlement of larvae in nature and under laboratory conditions. Three methods were used to registrate the tidal dependent dynamic of planulae release: 1) plankton collecting bags around Fucus inflatus kelp with mature hydroids colonies; 2) active stirring kelps with hydroids in container with water, which is an old way to stimulate planulae release; 3) direct account of the mature planulae into gonangia. The dynamic of intensity of L. flexuosa planulae release was investigated according 3-4 phases of tidal cycle. All data were statistically tested. For L. flexuosa a moment of general larvae release was found in phase with the period of low water. This correlation could explain strict limitation in occurrence of L. flexuosa only in the lower part of intertidal zone. Laboratory experiments show that planulae release is stimulated by littoral drainage, and renewal of water movement during the beginning of tide. The decrease in time of planulae settlement is an affective way for marine sedentary species to stay in a narrow zone of optimal habitat. PMID- 11881216 TI - [Some regularities in the development of encrusting colonies of Cribrilina annulata (Fabricius, 1780)]. AB - The growth of encrusting colonies was studied with mathematical model. It was shown that encrusting growth takes place under increasing competition for the substrate inside the colony. The model was tested on the example of Cribrilina annulata collected in White Sea on Laminaria saccarina. All colonies were mapped, zooids were measured and genealogical connections between them were established. A number of gradients were revealed by statistical methods. The intensity of budding decreases in astogeny according to theoretical predictions. It was shown that development of Cribrilina annulata colonies is strictly determined by gradients that can be caused by shortage of substrate space. It leads to the suppression of budding and changing in zooid size. Increasing substrate shortage is predictable and caused by the regularities in of zooid budding. The growth of colony stops after exhausting of potentially available substrate. PMID- 11881217 TI - [Analysis of possible mechanisms of regulation of root branching]. AB - The formation of lateral roots under the influence of growth inhibitors and phytohormons were studied with germs of corn and flax. All corn primordiums develop into the lateral roots without dormancy period, while some flax primordiums are delayed in development. Removal of root apical meristem (decapitation) does not result in the increase in number of lateral roots of corn germs. Decapitation of flax germs induces the development of some dormant primordiums, that leads to the considerable increase in number of lateral roots. Formation of additional primordium under decapitation does not take place. Auxin stimulates the formation of additional primordiums in flax, but does not effect the number of mature lateral roots. In corn germs synthetic auxin stimulates the development of additional primordiums in root zone, where primordiums are forming after the disturbance. It is shown that the time of primordium development does not change under inhibitor influence, if they do not selectively effect mitoses. It seems that auxins regulate initiation of pericycle division and formation of primordiums of lateral roots on the stage of development of meristematic protuberance. Cytokinins influence the development of already existed primordiums of lateral roots but do not effect the formation of new ones. Constancy of time of primordium formation and independence of their development on the state of the main root need future investigations. The authors suppose that regulation of these processes is non-hormonal. PMID- 11881218 TI - [Systematics on the threshold of the XXI century. Traditional principles and fundamentals from today's point of view]. AB - Systematics as regarded is a purely theoretical domain of biology, and its product, system, as a specific biological theory, or a topologo-genetic model of the biota. Linnaeus was the first to introduce the idea of system and the systematic approach into the natural history. The advent of evolutionism brought new meaning to the old term "affinity", so Linnaeus' slogan of natural system got new life, and Linnaeus taxonomy assimilated the evolutionary ideology quite naturally and much easier than many other departments of biology. The difference between natural and artificial systems is remaining, and it is in their goals, as formulated by Linnaeus: heuristic of the former and cataloguing of the latter. Linnaeus' clairvoyance discovered the existence of an infrageneric level of genetic integration provable by naturalists' experience. He chose for it the designation of "species" and laid it down as primary, basic unit of his system. This is plainly evident from his own writings; the story about Linnaean species being products of a logical division of genera is a pure fiction. Modern populational model of species, by 3 important criteria, appears to be more akin to the Linneaean one than to the ideas of Lamarckism and early Darwinism. Systematic approach focuses rather on the interrelations among elements and their relative position, then on the properties and qualities of separately treated individual elements. In the development of systematics the aspect of "nexification" (study of connections) has been continuously gaining attention especially regarding the nomenclature where connotation has been totally forced out by denotation. PMID- 11881219 TI - [The anthrax bacterium--a biological masterpiece with deadly potential]. PMID- 11881220 TI - [Abdominal sarcomas--rare tumors requiring multidisciplinary approach. Evaluation of surgical management in a sarcoma team]. AB - Retroperitoneal sarcomas and gastrointestinal stromal tumors are rare malignant tumors with a high recurrence rate. Due to their localization, and to the fact that they give rise to non-specific symptoms, these tumors are usually large at diagnosis. From 1990 to 1998 thirty-one patients (16 men and 15 women, mean age 45 years; range 15-77 years) went through surgical treatment. Patients were evaluated by a team consisting of an oncologist, a pathologist, a radiologist and a surgeon both pre- and postoperatively. Forty-five operations were performed (23 for primary tumors and 22 for recurrences). The resectability and radical resection rate were 89% and 59% respectively in primary tumors (4 patients with benign histological diagnoses were excluded) and 68% and 47% respectively in local recurrences. Other organs were removed in 50% of the procedures. Vascular and orthopedic surgeons participated when necessary. Morbidity and in-hospital mortality were 18% and 7% respectively. The overall recurrence rate was 70%. It is our belief that a multidisciplinary approach is mandatory for optimal treatment in these patients. PMID- 11881221 TI - [Free vaccine and increased information suggested to increase the vaccination coverage. A questionnaire study concerning influenza vaccination of elderly persons and other risk groups]. PMID- 11881222 TI - ["A wheelchair virus"--influenza caused severe pain of the calf muscles among children]. PMID- 11881223 TI - [Blood component transfusion in children. Current recommendations]. PMID- 11881224 TI - [A study of 56-65 years old persons in Molnlycke. No association between Helicobacter and heart disease or thyroid disorder]. PMID- 11881226 TI - [Dialysis is life-sustaining--but when to initiate it?]. PMID- 11881225 TI - [Diagnosis of depressive disorder and so-called exhaustion depression. Self esteem--a central concept]. AB - Depressive disorders can be recognized by the loss of self-esteem; this contrasts with mourning and neurasthenic reactions, in which self-esteem remains intact. Just as depression can result from the gradual reduction and eventual loss of self-esteem, mourning and neurasthenic reactions can evolve into true depressive states. "Exhaustion depression", a new diagnostic category connected to "burnout" situations, should be applied only when criteria for depressive disorder are fulfilled, including loss of self-esteem. When these criteria are lacking we should refer only to an exhaustion state provoked by stress. Neurotic mechanisms may represent a special class of relevant stress factors, but are not seen in manifest neurasthenic reactions and exhaustion depression. PMID- 11881227 TI - [A case report. FDG-PET in occult breast cancer]. PMID- 11881228 TI - [Alexander Malmquist--destiny of an artist. Misery and disease put a stop to a promising career]. PMID- 11881229 TI - [Magic and natural products features in Lapp traditional medicine]. PMID- 11881230 TI - [Phenomenological perspectives of health and disease]. PMID- 11881231 TI - [Increased freedom for general practitioners and primary health specialists in Stockholm]. PMID- 11881232 TI - [Documented effect of methadone, but not naltrexone]. PMID- 11881233 TI - [Methadone and naltrexone in heroin addiction--an explanation from the SBU]. PMID- 11881234 TI - [Is there an effective treatment of refractory angina pectoris?]. PMID- 11881235 TI - [The risk of hospital infection as an argument for home transportation of patients with fractures acquired abroad]. PMID- 11881236 TI - [Fatigue syndrome--a new name of a well-known psychological crisis reaction]. PMID- 11881237 TI - [Reduced mortality of intensive care patients by insulin therapy. The effect is especially evident in connection with heart surgery]. PMID- 11881238 TI - [Decreasing mortality of children with congenital heart abnormalities. A new population of adult patients requires increased resources]. PMID- 11881239 TI - [Congenital heart defects. Varying degrees of severity; 25 percent of children with heart defects will have persistent cardiac problems in adulthood]. AB - In a population based study including 35,218 infants born alive during the 15 year period 1982-1996, 360 (1%) were diagnosed having a congenital heart defect (CHD). At a follow-up 3-18 years later (median 9.5 years) 154 patients (42.8%) were spontaneously cured, of whom 142 (92.2%) had ventricular septal defects (VSD). 42 patients (11.7%) had died, 22 of whom (52.4%) during the neonatal period (0-28 days after birth). A total of 119 patients (33.1%) underwent therapeutic procedures (surgery, catheter interventions), of whom 24 (20.2%) died. Of the 95 children surviving therapeutic procedures 54 (56.8%) had their defects completely repaired, while 41 (43.2%) had residual defects or cardiac sequelae, often of minor importance. Of 69 children (19.2%) with persistent untreated defects, 43 (62.3%) had VSD. A chromosomal disorder, various syndromes or extracardiac malformations occurred in 72 children (20%). The study underlines the fact that CHD presents itself in varying degrees of severity, including a high neonatal mortality rate as well as a high rate of spontaneous cure. PMID- 11881241 TI - [Mouth dryness--invisible problem]. PMID- 11881240 TI - [Growing waiting list for corneal transplantation]. AB - New corneal storage techniques and legislative changes have transformed conditions for corneal grafting in Sweden. Since 1995, the Swedish Corneal Surgery Society has registered the number of grafts, the size of waiting lists for surgery as well as changes in the capacity of the banks. Since 1995 the number of procedures per year has declined while the waiting list has grown. The required number of procedures is 650 rather than the 448 performed in 1999. The decline is due to a lack of donor corneas, priority accorded to other surgery, efforts to adapt contact lenses to keratoconus, and further indications for excimer laser surgery. The four established corneal banks as well as two opening in 2001 will increase the number of grafts available for surgery. PMID- 11881242 TI - [Report of the status and presentation of knowledge sources. Infections in pregnant women, fetuses and newborn infants]. PMID- 11881243 TI - [Compression dressing after hip joint replacement reduces the need of allogeneic blood transfusion]. AB - Allogeneic blood transfusion may increase the risk for postoperative infections, allergic reactions and transmission of diseases. The aim with the present study was to evaluate the effect on blood saving with a new type of compression dressing after hip joint replacement surgery. 50 patients undergoing after hip joint replacement surgery were prospectively allocated to postoperatively get a standard dressing (n = 24) or the new type of a compression dressing (n = 26). The peroperative bleeding was similar in both groups. The need for allogeneic blood transfusions were significantly higher in the group of patients that got standard dressing. This study indicates that the compression dressing reduces the need for allogeneic blood transfusions after after hip joint replacement surgery. PMID- 11881244 TI - [Increasing interest in cold therapy]. AB - Hypothermia was used early in the treatment of malignant tumors. New evidence shows that hypothermia protects normal tissues from the toxic effects of radiation and chemotherapeutic drugs. Hypothermia in oncological research is calculated to increase the therapeutic ratio by protecting normal tissue while retaining the antineoplastic effect. Animal results indicate that removal of the primary tumor using cryosurgery diminishes metastatic tumor growth more than does surgical excision. PMID- 11881245 TI - [Treatment in the community health centers in accordance with recommendations of the Medical Products Agency. Unsatisfactory weight reduction with orlistat]. AB - We treated 44 individuals, 31 women and 13 men, for 12 months; each one had a body mass index > or = 28 kg/m2. Mean age was 53 years (range 20-75 years). Each individual visited a nurse regularly for diet recommendations, and each was provided a prescription for orlistat from his or her own doctor. The target weight loss of 2.5 kg prior to treatment with orlistat was obtained by 28 patients. After 3 months the average weight loss was 3.3 kg, and after 6 months, when 10 women and 6 men remained, the average weight loss was 6.1 kg and 6.5 kg respectively. The average weight decrease between 6 and 12 months was 0.3 kg and 2.7 kg for 7 women and 4 men respectively. Total cost for medical staff's working hours was approximately 700 Swedish crowns per kg weight loss. This cost seems rather high in comparison with the unsatisfactory results obtained for the group as a whole. PMID- 11881246 TI - [A case report. Rosiglitazone treatment was highly effective yet had to be terminated]. AB - The thiazolidinediones were introduced as oral hypoglycemic drugs in Sweden during the fall of 2000. A case is reported in which a woman with insulin dependent type-2 diabetes and both macro- and microangiopathy and pronounced insulin resistance was treated with rosiglitazone (Avandia). Within three months insulin doses could be reduced by 36% (from 176 to 112 units insulin daily) and concomitantly Ery-HbA1c was reduced from 8.4 to 5.3%. In spite of this dramatic effect on glucose homeostasis administration of the drug had to be discontinued due to critical congestive heart failure. PMID- 11881247 TI - [Fights and rows carried ophthalmology forward]. PMID- 11881248 TI - [Consequences of the Chernobyl accident are difficult to prove. Thyroid gland cancer among those who were exposed to radiation during childhood is the only disease evidently connected to the radiation so far]. PMID- 11881249 TI - [A myth that organizational changes have a rational basis. A report on and thoughts about dissertation]. PMID- 11881250 TI - [A direct question: Is orthorexia a correct word for a wrong concept?]. PMID- 11881251 TI - [Keep and improve the orientation course on psychotherapy!]. PMID- 11881253 TI - [Is psychiatrists' knowledge of psychotherapy discriminatory?]. PMID- 11881252 TI - [The compulsory psychotherapy course is dead: long live psychotherapy!]. PMID- 11881254 TI - [Learn from the county of Blekinge!]. PMID- 11881255 TI - [Keep the compulsory course on psychotherapy--reform the psychotherapeutic internship training]. PMID- 11881256 TI - [Absolutely necessary education for future psychiatrists]. PMID- 11881257 TI - [Are Swedish psychiatrists to become "half-educated scientists" again?]. PMID- 11881258 TI - [Keep the compulsory course--suggestion to measures]. PMID- 11881259 TI - [Basic courses and graduate courses--are these two never going to meet?]. PMID- 11881260 TI - [The discussion on breast feeding and childhood cancer in mass media has evoked anxiety among parents]. PMID- 11881261 TI - Shaky ground. Deteriorating system jeopardizes state's mentally Ill. PMID- 11881262 TI - Evaluating prenatal care in Arkansas. PMID- 11881263 TI - Evaluating suspected pulmonary embolism. PMID- 11881264 TI - Case of the month. Symptomatic chronic calcified subdural hematoma in a 5-year old child. AB - Head trauma is a frequent cause of morbidity and mortality in the pediatric population. The pathophysiology and clinical outcomes of head trauma differ between children and adults. Traumatic mass lesions such as subdural and epidural hematomas occur less frequently in children and, when present, are associated with lower mortality. Subdural hematoma is the collection of blood on the cortical surface beneath the dura with bleeding from bridging veins or cortical arteries. These hematomas are usually associated with trauma. Chronic subdural hematomas are much more common in infants and frequently exist as a single entity; it is rare for chronic subdural fluid accumulations to occur after one year of age. Specific traumatic events are usually unrecognized or unreported. In rare instances, subdural hematoma may indicate an underlying bleeding disorder, hematological malignancy or benign expansion of subarachnoid space. We report the dramatic presentation of a strikingly large calcified chronic subdural hematoma in a 5-year-old with increased intracranial pressure and subfalcine herniation. PMID- 11881265 TI - Should family members be present during CPR? AB - It has been debated recently whether or not relatives should be allowed presence during the attempted resuscitation of their loved one. Medical professionals from 81 hospitals in Arkansas were surveyed, and 223 surveys were returned. Out of these, 85 respondents said they would consider allowing family member presence during CPR, while 131 respondents said they would not consider allowing family presence. The most common reason given for not allowing family member presence was that the family was unprepared or that the CPR scene was too traumatic for family presence. However, other studies have shown that with preparation and support, some family members are helped in their grieving process by being allowed presence. Medical paternalism is not a valid reason for the routine exclusion of families from the resuscitation room. PMID- 11881266 TI - Sensation and Distress of Pain Scales: reliability, validity, and sensitivity. AB - Psychometric properties of the Sensation and Distress of Pain Visual Analogue Scales (VAS) are compared to dual numerical rating scales (NRS) with data from a randomized controlled trial of postoperative patients. On postoperative days 1 and 2, 15-minute test-retest reliability was .73 to .82 for the VAS and slightly lower for the NRS, r = .72 to .78, while convergent validity of the scales ranged from r = .90 to .92; construct validity of sensation and distress ranged from r = .72 to .85; and discriminant validity was lower, r = .65 to .78. Both instruments were significantly associated with pain reduction following treatment, p < .05 to .01. The VAS scores were significantly lower, p < .01 to .001, and more evenly distributed than NRS scores. It is recommended that the VAS be used in research to produce continuous scores that are more suited to parametric analysis. PMID- 11881267 TI - Family Caregivers' Motives for Helping Scale derived from motivation-to-help theory. AB - The literature does not contain a measure of family caregivers' motives for helping provide daily complex home care. Such a measure will permit nurses to assess and provide interventions specific to each caregiver's motivation for helping. The purpose of this study was to apply Batson's empirically derived helping pathway theory to the measurement of caregiver motives for helping and develop a short form that does not add to the burden of caregiving. A Principal Components factor analysis (N = 93) of Family Caregivers' Motives for Helping Scale is used. Criterion-related validity is ascertained using a triangulated, independent validation procedure with qualitative data from a subsample (N = 41). Subsample subjects' interview data were categorized, based on Batson's theoretical pathway definitions, by coders blinded to caregiver Motives for Helping Scale scores. Three of Batson's four helping pathways (reward seeking, altruism, and punishment-avoidance) were extracted during factor analysis. This three-factor solution explained 66.6% of the variance and was confirmed by a 97% agreement between three of Batson's pathways and caregivers' helping motive score. The content analyses of the descriptive interview data also coincide with the 3-factor solution. The scale items representing Batson's fourth helping pathway, distress reaction, were not retained due to cross loading. The Family Caregivers' Motives for Helping Scale accurately measures three of four theoretically derived motivations for helping another. The scale should be reanalyzed in a larger sample of caregivers. Aligning nursing interventions to caregiver motives for helping can provide reinforcement for caregivers and potentially enhance home care outcomes. PMID- 11881268 TI - Development of an exercise self-efficacy questionnaire for older Mexican American women. AB - The purpose of this study was to develop a reliable and culturally meaningful exercise self-efficacy questionnaire for older Mexican American women. Items were derived from a 5-item exercise self-efficacy instrument (Laffrey, 2000; Marcus, Selby, Niaura, & Rossi, 1992) and from focus groups with women in the target population. Content validity of the Spanish-language items was supported by three Mexican American professional women and by five Mexican American nurses. A factor analysis of the 11-item questionnaire revealed one major factor. Internal consistency was shown by an alpha reliability coefficient of .86. Item:corrected scale correlation coefficients ranged from .30 to .70. The mean exercise self efficacy score was significantly higher for 38 regular exercisers than for 37 non exercisers. The exercise self-efficacy questionnaire can be used in research with older Mexican American women and should be tested with men and with younger men and women to explore its usefulness with these populations. PMID- 11881269 TI - Measurement of activity in older adults: reliability and validity of the Step Activity Monitor. AB - The purpose of this study was to test the reliability and validity of the Step Activity Monitor (SAM) when used with older adults. A total of 30 subjects with a mean age of 86 +/- 6.1 participated in the study. Sixty one-minute walks were measured with the SAM, and two observers visually counted steps. Four participants wore the SAM for 6 to 48 hours and maintained activity diaries. The intraclass correlation for the SAM recordings was R = .84. There was an overall step counting accuracy of 96%. The diaries supported the SAM data for those who wore the SAM for extended periods. The SAM is an easy to use, comfortable, valid, and reliable measure of activity in older adults and particularly may be useful to triangulate measurement of activity in these individuals. PMID- 11881270 TI - Psychometric assessment of the Adult-Adolescent Parenting Inventory in a sample of low-income single mothers. AB - The Adult-Adolescent Parenting Inventory (AAPI) is a 32-item inventory widely used to identify adolescents and adults at risk for inadequate parenting behaviors. It includes four subscales representing the most frequent patterns associated with abusive parenting: (a) Inappropriate Expectations; (b) Lack of Empathy; (c) Parental Value of Corporal Punishment; and (d) Parent-Child Role Reversal. Although it has been used in a variety of samples, the psychometric properties of the AAPI have not been examined in low-income single mothers. The purposes of this study were to: (a) examine the reliability and validity of the Adult-Adolescent Parenting Inventory (AAPI) in a sample of 206 low-income single mothers; (b) assess the mother's risk for inadequate parenting by comparing their AAPI subscale scores with normative subscale scores on the AAPI; (c) assess the construct validity of the AAPI by testing the hypothesis that mothers with lower AAPI scores have a higher level of depressive symptoms and lower self-esteem in comparison to mothers with higher AAPI scores; and (d) determine whether the 4 factor structure proposed by Bavolek (1984) could be replicated. AAPI scores indicated these mothers were at high risk for child abuse when compared with normative data for parents with no known history of abuse. Higher risk for abusive parenting was associated with a higher level of depressive symptoms, less education, and unemployment. The subscales, Inappropriate Expectations and Parental Value of Corporal Punishment demonstrated poor internal consistency with Cronbach's alphas of .40 and .54, respectively. Hypothesis testing supported the construct validity of the AAPI. Bavolek's 4-factor structure was not supported. A 19-item modified version of the AAPI with three dimensions was identified. This modified version of the AAPI may provide a more efficacious tool for use with low income single mothers. PMID- 11881271 TI - Psychometric properties of the California Critical Thinking Tests. AB - The evaluation of critical thinking, as with any other measure, must employ instruments that meet appropriate psychometric standards. Therefore, the purpose of this study was to investigate the psychometric properties of the two California Critical Thinking Tests. Two samples of undergraduate students enrolled in a southern comprehensive university took the California Critical Thinking Skills Test (TST) and the California Critical Thinking Dispositions Inventory (TDI). A portion of the participants, who were nursing students, were retested on the TST and TDI two weeks after the initial testing. Some participants also completed the Watson-Glaser Critical Thinking Appraisal (WGCTA). In both samples the TST had neither sufficient psychometric properties to assess individual abilities nor sufficient stability reliability. In particular, the analysis subscale of the TST demonstrated exceptional weakness in all administrations. In contrast, the TDI demonstrated very good reliability estimates in all administrations of the test. The data did not conform to the subscale structure during factor analysis. Whereas the TST needs further refinement, the TDI demonstrated sufficient reliability for use in attitudinal research. PMID- 11881272 TI - The lab's role in developing an institutional drug testing policy. PMID- 11881273 TI - Talking with Kenneth Freeman of Quest Diagnostics. Interview by C. Anne Pontius. PMID- 11881274 TI - Prenatal screening tests facilitate risk assessment. AB - Medical technology continues to improve services and testing to provide accurate and timely information for physicians and patients. New screening protocols in which information obtained in the first and second trimesters is integrated to improve detection and lower false positive rates have potential for safer and more effective prenatal care. Families still have difficult choices to make when confronted with the diagnosis of a birth defect. Inherent advantages of early detection are reassurance for low-risk women sooner, and for women with increased risk, more time to consider all options. Without doubt, improvements in risk assessment, diagnosis, and prevention will continue. The advantage offered by laboratory procedures is that the preliminary diagnosis can be made by simple, noninvasive screening tests. PMID- 11881275 TI - [Changes in serum levels of lipid peroxidation products during labor and in the puerperium]. AB - OBJECTIVE: To determine the activity of reactive oxygen species (ROS) during labour by characterizing changes in maternal serum levels of lipid peroxidation end-products--MDA and lipofuscin during labour and the early post-partum period. We also tried to evaluate the relationship between levels of lipid peroxides and some clinical characteristics of labour. DESIGN: Prospective study. SETTING: Department of Obstetrics and Gynaecology LFUK, Bratislava; Department of clinical laboratories, Ministry of defense SR, Bratislava. METHODS: Blood samples were obtained from 66 pregnant women with uncomplicated pregnancy at the end of labour and during the early post-partum period. The control group consisted of 19 pregnant women delivering by primary Cesarean section. Blood samples were examined for MDA and lipofuscin by HPLC method. We used paired and unpaired Student's t-test to statistically evaluate our results. RESULTS: MDA and lipofuscin levels in pregnant women delivering spontaneously compared to those delivering by C-section were significantly elevated (P < 0.05). MDA and lipofuscin levels in pregnant women during spontaneous labour or during by C section compared to the levels in early post-partum period were not significantly increased. We have not found any correlation between the length of the labour and lipoperoxides concentration. PMID- 11881277 TI - [Growth and development in untreated girls with slowly progressing idiopathic central or borderline-onset puberty]. AB - OBJECTIVE: To analyse growth and development of girls with slowly progressive idiopathic precocious or early puberty. DESIGN: Long-term open clinical study. SETTING: Department of Obstetrics and Gynaecology, Second Faculty of Medicine, Charles University, Prague. METHODS: In 20 untreated girls with slowly progressive puberty starting at 6-9 years neurogenic aetiology was excluded. During follow-up period 4.7 +/- 2.2 (2-8.5) years (mean +/- SD; range), sexual development (Tanner criteria), age at menarche, menstrual cycle and auxological parameters were evaluated. RESULTS: 13 girls reached menarche at 11.1 +/- 0.9 years (3.7 +/- 1.1 years after the onset of puberty), earlier than in their mothers (12.9 +/- 1.1 years) and Czech standards (P < 0.05). Menstrual cycle 28 (24-29) days was regular in all 6 girls with gynaecological age > 2 years. In one girl microprolactinoma was diagnosed, therapy with bromocryptine started at the age 14.7 years (3.5 years after menarche). At the onset of follow-up, bone age (TW20) advancement was 1.8 +/- 1.4 years above the chronologic age. Initial prediction of final height (graphic method) was 162.3 +/- 5.5 cm vs final prediction 163.7 +/- 5.1 cm. Final height 162.2 +/- 5.7 cm achieved 7 girls vs target height 163.6 +/- 5.2 cm (NS). CONCLUSION: In untreated girls, menarche occurred later after the first signs of puberty than in normal population, menstrual cycle was regular. Height potential was preserved, final height corresponded with their target height. Not all girls with early and slowly progressive puberty should be treated. Therapy is necessary in organic aetiology, rapid progressive precocious puberty and impaired growth prognosis. PMID- 11881276 TI - [Serum levels of leptin and soluble leptin receptors in women after bilateral ovariectomy]. AB - OBJECTIVE: To determine whether the blood concentrations of leptin and soluble leptin receptor (LEPR) hang in women after bilateral ovariectomy, and whether there is a relationship between the concentrations of leptin, LEPR and LH, FSH and 17 beta-estradiol before and after castration. DESIGN: Prospective clinical study. SETTING: Department of Obstetrics and Gynaecology, First Faculty of Medicine, Charles University and the General Faculty Hospital, Prague. METHODS: 10 women of reproductive age (31-37 years) with a regular menstrual cycle were included in the study. Blood samples were always taken during the follicular phase of the menstrual cycle and 5 days after bilateral ovariectomy. In 9 women, hysterectomy was performed with the bilateral ovariectomy. Evaluated parameters: leptin, soluble leptin receptor, FSH, LH, 17 beta-estradiol and BMI. Serum soluble leptin receptor and leptin concentrations were determined using commercial ELISA kits. Serum concentrations of FSH, LH, and 17 beta-estradiol were determined using commercial RIA kits. RESULTS: We did not find any statistically significant changes in the concentration of leptin (8.9 +/- 5.5 vs 7 +/- 5.6) and LEPR (16.2 +/- 5.6 vs 20.9 +/- 8.9) before and after surgical castration. After surgery, according to expectations, the concentration of LH and FSH significantly increased while the concentration of 17 beta-estradiol decreased (P < 0.05). Five days after surgical castration we found only a tendency toward decrease in the concentration of leptin and an increase in the LEPR concentration. The differences were not statistically significant. Except for the positive correlation between leptin concentration and BMI, we did not find any statistically significant correlation between the concentrations of leptin and LEPR and any of the other evaluated parameters in both groups. CONCLUSION: In our study, there was a significant decrease in 17 beta-estradiol concentration five days after surgery. In contrast, the concentrations of FSH and LH significantly increased after surgery, which is in agreement with the expected endocrinological changes after ovariectomy. There was a tendency toward a decrease in serum concentrations of leptin and increase in serum concentration of LEPR after surgery, but it was not statistically significant, which is in contrast to another study published regarding this topic. PMID- 11881278 TI - [Supplemental imaging methods in evaluation of the extent of cervical carcinoma]. AB - OBJECTIVE: To present a review of literature and to analyse of contribution of imaging methods in staging of cervical carcinoma. SUBJECT: Review. SETTING: Department of Obstetrics and Gynaecology, Medical Faculty Trnava, Slovak Republic. SUBJECT AND METHOD: Discussion about relevant evidence from literature, information database (MEDLINE) review, and results from own prospective study. CONCLUSION: FIGO staging of cervical carcinoma is based on clinical examinations but is complemented by imaging methods to optimalize treatment planning. From used imaging methods (computed tomography, magnetic resonance imaging, lymphography, sentinel node mapping, ultrasonography) is combination of abdominal and endoluminal ultrasonography the best available and affordable method with significant benefit in decision making procedure. PMID- 11881279 TI - [Role of sperm antibodies and cellular autoimmunity to sperm in the pathogenesis of male infertility]. AB - OBJECTIVE: To test the hypothesis of relationship between sperm pathology and elevated humoral and/or cell-mediated antisperm autoimmunity in male partners from infertile couples. DESIGN: Analytic study. SETTING: Department of Immunobiology, Institute for the Care of Mother and Child, Prague. METHODS: Sperm samples were evaluated according to WHO rules. Sperm-bound antisperm autoantibodies (ASA) were determined by SpermMar Test (FertiPro N. V., Sint Martens-Latem, Belgium). For evaluation of cell-mediated antisperm autoimmunity (CMAA) the authors used their own modification of migration-inhibition test (Dimitrov et al., J. Immunol. Methods 154: 147, 1992). RESULTS: The pool of men was divided into groups according to the result of sperm examination: normozoospermia (740 men), asthenozoospermia (244), teratozoospermia (191), oligoasthenozoospermia levis (61), oligoasthenozoospermia gravis (29), oligoteratozoospermia (82), and azoospermia (54). Subgroup of fertile men (32) consisted of normozoospermic men--fathers of child younger than 3 years. Percentage of sperm-bound ASA-positive samples was significantly higher in asthenozoospermia in comparison with normozoospermia in both IgA (20.8% versus 10.6%) and IgG classes (13.8% vs 6.8%). Positivity of CMAA was significantly more frequent in group of asthenozoospermic (52%) than in normozoospermic (28.5%) and fertile (12.5%) men. CONCLUSION: Antisperm autoimmunity, namely its cell-mediated form, appears to play a significant role in impairment of spermiogenesis. Sperm bound autoantibodies were found more frequently in asthenozoospermia, but also in some men with normozoospermia they may impair fertility. PMID- 11881280 TI - [Massive pulmonary embolism after delivery by cesarean section]. AB - OBJECTIVE: To give an overview of the preventive and therapeutic measures in thromboembolic disease in association with pregnancy and delivery. SUBJECT: Case report. SETTING: Department of Obstetrics and Gynaecology, First Faculty of Medicine, Charles University and General Faculty Hospital, Prague. SUBJECT AND METHOD: The surgical treatment of massive pulmonary embolism in a patient after delivery by caesarean section. CONCLUSION: Surgical embolectomy still has its place in the treatment of pulmonary embolism in the early phase of critical cases, when thrombolysis is contraindicated, and mechanical disintegration with the catheter is unsuccessful. In such cases it is the only one possibility of saving the patient. However, prevention of thromboembolic disease remains of primary importance. PMID- 11881281 TI - [Laparoscopy in chronic pelvic pain--a retrospective clinical study]. AB - OBJECTIVES: To analyze the laparoscopic findings in women with chronic pelvic pain (CPP). To verify the possible predictive value of the anamnestic factors for the laparoscopically established diagnosis. DESIGN: A retrospective clinical trial. SETTING: Department of Obstetrics and Gynaecology, 1st Faculty of Medicine and the General Faculty Hospital, Charles University, Prague. METHODS: We realized a detailed analysis of laparoscopies for CPP performed in our clinic in the last 5 years. With the help of statistical analysis (chi 2) the possible influence of age, previous surgical intervention, history of PID, dysmenorrhea, infertility, and some other factors on endoscopic finding was verified. RESULTS: 480 laparoscopies for CPP were performed from the year 1995 to 1999. The most frequent findings were adhesions (22.3%), endometriosis (20.4%), PID (17.7%), and normal finding (17.7%). In 53% of all cases the operative laparoscopic procedure was done, most frequently the adhesiolysis (62.2%). Only one serious complication during laparoscopy was recorded (0.21%). In women younger than 30 years endometriosis was the most frequent finding (22.8%), in women older than 30 pelvic adhesions were the most often (31.9%). Diversity of findings between the groups was statistically significant (P < 0.0001). In patients with previous operation in pelvis the adhesions were found most frequently (46.2%). Diversity of findings in women with and without previous operation was also significant (P < 0.0001). In women treated for PID, pelvic inflammation was found in 25.8%, but in 22.5% the finding was negative and in 20.2% the endometriosis was diagnosed. In these subgroups (women after and without treatment of PID) the diversity of findings was not significant (P < 0.1). In infertile women, suffering from CPP, PID was found most frequently (41.2%). A suspicion of chronic appendicitis was verified in 64.3%. In patients with dysmenorrhea, as well as in women suffering from dyspareunia, endometriosis was dominant finding (30.4%, resp. 29.1%). CONCLUSION: Laparoscopy for CPP is a safe and effective method for verifying and adequate therapy of as yet hidden pathological findings. With the help of anamnestic factors some laparoscopic findings are predictable; although the sensitivity and specificity is very poor. PMID- 11881282 TI - [Acebutolol in the treatment of arterial hypertension in pregnancy--comparison with commonly used hypertensive agents]. AB - OBJECTIVE: The main purpose of our study was to analyse efficacy and safety of acebutolol (Sectral) in the treatment of hypertension in pregnancy. DESIGN: Cohort clinical study comparing efficacy and safety of acebutolol with other antihypertensive drugs by 77 patients and their 81 newborns. The number of 48 patients were treated by acebutolol. SETTING: Internal Medicine and Clinical Pharmacology Department, Regional Hospital, Nitra and The Faculty of Health Service and Social Work, University of Trnava; Department of Gynaecology and Obstetrics, Regional Hospital Nitra and The Faculty of Health Service and Social Work, University of Trnava; Institute of Pharmacology, The Faculty of Medicine, Komenskeho University, Bratislava; Environment, a.s. Nitra. METHODS: During the period of 6 years our patients were divided into subgroups according to the type of hypertension in pregnancy and the severity of the illness which was the basis for used antihypertensive drugs. One of the topics of the study was to provide monitoring of adverse events by mother and possible drug influence on their new born has been evaluated by investigating the week of birth, weight and length of the new-born and evaluating of Apgar score. RESULTS: In the subgroup of 48 women treated by acebutolol we have confirmed the efficacy and safety of this antihypertensive drug without any clinically significant effect on the quality of life of their new-born and evaluating of Apgar score. CONCLUSION: With regard to the results of our clinical study we can consider acebutolol to be effective and safe antihypertensive drug in the treatment of hypertension in pregnancy. PMID- 11881283 TI - [Organ torsion and abdominal symptoms--diagnosis]. AB - The authors reflect on the etiology of torsions of intraabdominal organs and the testes. They mention anamnestic data, which may lead to suspicion of possible organ torsion. They analyze the clinical picture of a patient who suffered such an attack. The authors submit a list of diagnostic possibilities using modern imaging methods: sonography, computed tomography, angiography, scintigraphy and magnetic resonance. They discuss laparoscopy as a diagnostic and therapeutic method. PMID- 11881284 TI - [Organ torsion and abdominal symptoms--case reports]. AB - Torsions are rare acute abdominal conditions and are mistaken for other more frequent diseases. The present work draws attention to the most frequent diagnostic errors. The authors present three cases of torsions of intraabdominal organs and two cases of testicular torsion. All patients attended their doctor because of abdominal pain. In four of five cases the patients were first treated for an erroneous diagnosis of acute abdomen. In the first case the torsion of the omentum was mistaken for diverticulitis of the sigmoid, later for an intraperitoneal lipoma, in he second case for cholecystitis, in the third case a patient with torsion of a myoma was indicated for surgery on account of acute appendicitis. In the fourth case incomplete torsion of the testis was mistaken for irritation of the appendix. In the fifth case where abdominal symptomatology dominated the correct diagnosis of testicular torsion was made and atypically spontaneous detorsion of the testis occurred. PMID- 11881285 TI - [Pedal bypass in the treatment of ischemic diabetic foot]. AB - AIM: The aim of this retrospective analysis was assessment of both patency and limb salvage rates of diabetic ischemic feet which underwent pedal bypass surgery within the period of 3 years. MATERIAL AND METHODS: Authors evaluated 50 critically ischemic diabetic feet where the pedal bypass was indicated for limb salvage. Total 41 pedal bypasses were performed and median follow-up was 16.7 months (1-39). The free muscle transfer were performed either in one session with or subsequently after the pedal bypass surgery in six limbs. RESULTS: Authors achieved 61% and 76% graft patency and limb salvage rates, respectively, within the median 16.7 months follow-up. All successfully revascularised feet healed up within the median period of 3 months postoperatively. No patient died within 30 days postoperatively. CONCLUSION: We conclude, that pedal bypass is safe procedure with acceptable limb salvage rate in the medium-term follow-up. Surgeons can markedly extend an indication criterions for limb salvage by exploiting of this technique. PMID- 11881287 TI - [Thromboembolism--present status and perspectives]. AB - The author summarizes contemporary findings on the problem of thromboembolism. He discusses its incidence, economic costs associated with treatment of complications such as arterial and venous thromoembolism. He emphasizes the problem of new types of prevention, in particular pentasaccharides in relation to complications and to other possibilities of prevention and treatment. The results of recent international studies and perspectives in this field are presented. PMID- 11881286 TI - [Serum leptin levels in the perioperative period in surgery patients]. AB - AIM: To assess dynamics of serum leptin concentrations (as a possible marker of stress) at some surgical patients during short perioperative period. PATIENTS AND METHODS: The serum leptin concentrations were estimated in group of patients with trauma or large bowel diseases in stress of both own disease and next elective or urgent surgery. The subgroups of patients were: a) monotrauma (13), b) polytrauma (10), c) malignant (20) or d) inflammatory disease (13) of large bowel. The serum leptin concentrations were evaluated preoperatively and during 16 hours postoperatively-immediately after surgery, after 8 and 16 hours after operation. The serum IL-6 concentrations were estimated immediately and 48 hours postoperatively. RESULTS: The serum leptin concentrations showed a time depending dynamic phase: they decreased postoperatively and increased during the first 16 hours postoperatively. It was more expressed in the malignant bowel disease subgroup. A correlation between age and serum leptin concentrations was not found in any subgroup. We found correlation between serum leptin concentrations preoperatively and postoperatively. CONCLUSIONS: The serum leptin concentrations demonstrated some dynamics in short perioperative periods. It was very considerably in patients with large bowel malignancy. Any significant difference in serum IL-6 concentrations was not discovered preoperatively and postoperatively in all subgroups. PMID- 11881288 TI - [A complicated case of esophageal carcinoma]. PMID- 11881289 TI - [Carotid microendarterectomy in the aged]. AB - The authors evaluate the results of 100 carotid endarterectomies in 88 patients, 70 plus years old, operated between April 1997 and December 2000. Regional anaesthesia was always used. The use of intraluminal shunt was selective (11%), on the basis of consciousness and the neurological status. Operating microscope and microsurgical technique were used in all cases. Neither 30-d morbidity nor mortality has been detected in the group. Medical morbidity has been recorded in 2 patients. CONCLUSION: Carotid microendarterectomy in regional anesthesia represents effective procedure for patients in the old age. PMID- 11881290 TI - [Metastasis as the first sign of thyroid carcinoma]. AB - The authors present on the basis of case report the problematic of thyroid gland cancer where was manifest long-term (3 years) clinical only as the resistance on the right part of the neck in the trigonum submandibular. Histological examination confirmed the metastasis of thyroid gland papillocarcinoma. Total thyroidectomy was performed and the same type of cancer in both lobes was confirmed. The patient has been 2 years after surgery and radiotherapy (radioiodine) without recurrence. PMID- 11881291 TI - [Lymphoma of the small intestine]. AB - The authors analyze on a case-history the problem of lymphomas of the gastrointestinal tract. The patient was a female who was operated on account of a suspected perforation. On operation multiple perforations of the small intestine were found. A resection was performed. According to the histological results it was a giant-cell B-lymphoma of the small intestine. The postoperative course was without major complications. The patient was referred to the oncological department without further treatment. PMID- 11881292 TI - [Total tyroidectomy in malignant goiter, significance and problems]. AB - Surgical operation of the thyroid gland is a basic operation in the treatment of malignant diseases of thyroid gland. At present, total removal of the thyroid gland (total thyreoidectomy = TTE) has been the most frequently used means of the surgical intervention. TTE has been a unique surgical method for thyroid cancer. The aim our study was to confirm the increasing number of radical operations, how often surprising and unexpected histological findings of carcinoma appear, the incidence rate of carcinoma among the patients being operated on for goiter, which histological findings are most frequently encountered and the survival rate of the patients with carcinoma of the goiter. In our retrospective study, we have collected a set of 233 patients with goiter from 1997 until 1999. Nineteen patients from this group had thyroid cancer: 16 females, 3 males, average age 55.5 years. The histological findings represents 14 papillary carcinomas, 3 folicullar carcinomas, 1 medullary carcinoma and 1 malignant lymphogranuloma. TTE was performed in any case where there was suspicion of thyroid cancer before the operation. In the case of one sided operations, when we had no suspicion of thyroid cancer but histological investigation found cancer, we had to remove the opposite lobus (TTE). However, we have found an increasing rate of thyroid cancer. The unexpected histological finding of carcinoma was reported in 69% of the cases. The number of these cases was higher in the middle 2000 (83%). Clinical stage la, microcarcinoma, was found in 52.9% of papillary carcinomas, and further oncological treatment was not indicated. The number of proper radical operations has been increasing. The high rate of unexpected histological findings of thyroid carcinoma is the main reason for early indication for operation of goiter and the radical extent of the primary operation. PMID- 11881293 TI - [Ampulloma of Vater's papilla--case report]. AB - The authors discuss, based on a case-report, the diagnostic problems of malignities of the pancreas and ampulla of Vater. In the same patient they found an interesting complication after laparoscopic cholecystectomy (LCHE). It was a female patient who had LCHE four years previously where an ampulloma of the ampulla of Vater was diagnosed with invasion into the pancreas and a suspect secondary in the lower lobe of the right lung. On operation a pulmonary abscess was detected from a retained concrement. Duodenopancreatectomy which was performed did not reveal any tumour in the pancreatic area. PMID- 11881294 TI - [Antibodies to granulocytic Ehrlichiae in the population of the western and central part of the Czech Republic]. AB - A total of 963 subjects in four groups from western and central Bohemia were examined in 1998-2000 for the presence of antibodies against granulocytic ehrlichiae. In the indirect fluorescence assay, a specific antigen from ehrlichiae (HGE) cultivated in-vitro and commercial anti-IgG and anti-IgM secondary antibodies were used. An overall prevalence of HGE antibodies of the class IgG corresponding to titres < or = 1/80 and 1/160 was found to be 21.0 and 13.5%, respectively, with a predominance during the warm season (May-October). The seasonal seroprevalence was discriminated best from an all-year background by titres 1/80-1/160. Elevated antibodies were found more frequently in older subjects; no effect of gender was observed. Antibodies of the class IgM were detected only exceptionally and did not exceed the titre of 1/40. Only minor differences were observed in the prevalence of HGE antibodies between groups differing geographically or by the presence of Borrelia antibodies. The seasonal incidence of HGE antibodies did not differ significantly from that of Borrelia antibodies. PMID- 11881296 TI - [A water-borne epidemic of tularemia in Chlumcany]. AB - During an epidemic of tularaemia in Chlumcany in December 2000 48 people, 36 adults, 12 children fell ill. In the patients the oroglandular form predominated which was diagnosed 46 times. None of the patients died. The vehicle of the infection was inadequately treated water which was used by the patients for the preparation of soda water and for washing. Using the polymerase chain reaction francisellae were detected in two water samples. PMID- 11881295 TI - [Prevalence of antiborrelia antibodies in two populations: various interpretations of the same data]. AB - OBJECTIVE: The objective of the work was to test whether differently formulated hypotheses, when using correct statistical methods, and the same data lead to the same results. The work pertains to the interpretation of the prevalence of antiborrelia antibody levels in more or less high risk populations. GROUP: A group of 200 blood donors from Prague (100 men, mean age 33.5 years and 100 women mean age 24.4 years) and 71 subjects with a higher risk of contact with ticks infected with Borrelia burgdorferi--forestry workers from the Sumava mountains (39 men, mean age 40.6 years and 32 women, mean age 44.5 years). METHOD: In all participants antiborrelia antibodies class IgM and IgG were assessed and using the Western blot method the presence of specific antigens was examined. The blood samples of all probands were analyzed in the national reference laboratory for Lyme borreliosis CEM in the National Institute of Public Health in Prague. To test the hypothesis on the difference of groups as regards the frequency of positive antibody levels the chi square test was used. For testing the hypothesis on the difference of the groups as regards antibodies irrespective of normal laboratory values the non-parametric method--the Mann-Whitney test was used. RESULTS: The group of Prague blood donors and forestry workers from the Sumava mountains do not differ as to the individual frequency of positive antibody values assessed by the ELISA method in class IgM and IgG but they differ in class IgM assessed by the Western blot method. Both groups differ moreover markedly in the general distribution of antibodies unrelated to normal laboratory values assessed by the ELISA method in class IgM, in class IgG the distribution does not differ. CONCLUSION: We provided evidence that when using a different formulation of the hypothesis while using correct statistical methods identical data can be differently interpreted. Thus in the interpretation of the prevalence of antiborrelia antibody levels in healthy populations with a different risk--even if the two groups do not differs as to the frequency of positive findings, there is a significant statistical difference in their distribution. PMID- 11881297 TI - [Encephalitozoon cuniculi and Encephalitozoon intestinalis--causes of opportunistic infections]. AB - Microsporidia (phylum Microsporidia) are intracellular parasites that infect a wide range of protozoa, invertebrates and vertebrate hosts. Over a 1000 species have been classified into approximately 100 genera. Historically, microsporidial infections in silkworms, honey bees, and salmonid fish have been responsible for significant economic losses. More recently, microsporidiosis has been recognized as an important opportunistic infection in immunologically compromised patients. In this review there is information on the immunobiology of microsporidia Encephalitozoon cuniculi and Encephalitozoon intestinalis which were identified as the most common causative agents of microsporidiosis in mammals. Most of what is known about the immunology of microsporidiosis is based on experiments with the microsporidian Encephalitozoon cuniculi. PMID- 11881298 TI - [Detection of HIV antibodies in clients of the AIDS Center of the District Health Service in Karvina 1993-2000]. AB - The authors present the trends of some characteristics recorded in a group of subjects examined for HIV antibodies in the AIDS centre of the District Hygiene Station in Karvina in 1993 to 2000. These characteristics reflect the development of activities of the centre. During the period a total of 3733 HIV tests were made, incl. three positive ones. In two instances men with homosexual relations in the case-history were involved, in one case a male heterosexual. The declining trend of the ratio of anonymous examinations is apparent from 58.8% in 1993 to 7.7% in 2000. Parenteral drug users, prostitutes (in particular women) and homosexuals account on a long-term basis for ca 30% of all examinations. Recently this ratio is even higher. Repeated examinations (check-ups) of the same client account for 16.1% of all implemented HIV tests. At least once 384 parenteral drug users were examined, in prostitutes similarly as in men with homosexual relationships the number of subjects examined at least once is ca 200. Repeated tests were made in ca one quarter of all parenteral drug users, ca one third of the prostitutes and ca one fifth of homosexuals. In none of the repeated tests seroconversion to HIV positivity was recorded. To achieve close contact and monitoring of the HIV statute of groups with a risk behaviour despite their existence at the brink of law and despite their natural tendency to hide is real. Without collaborating organizations, without gaining their confidence and in particular without the confidence of the clients this would not be possible. PMID- 11881299 TI - [The VIII. Epidemiology Seminar]. PMID- 11881300 TI - [Characteristics of strains of Salmonella enterica serovar Typhimurium DT104 isolated in the Slovak Republic]. AB - From 1997 to 2000 in the Slovak Republic 8 strains of Salmonella enterica serovar Typhimurium DT104 were isolated from sporadic cases of human salmonellosis. Four strains were multiresistant and three strains were sensitive to antibiotics. We investigated the influence of the cultivation medium (complete Mueller-Hinton, synthetic-Staples) on growth, surface hydrophobicity and motility of the mentioned isolates. The hydrophobicity was evaluated by methods of adherence to the hydrocarbon xylene (BATH) in a salt-aggregation test with ammonium sulphate (SAT) and adherence to a plast. The growth of tested strains in 24 hours was greater after cultivation in the complete medium, but the salmonellae grew considerably also in the synthetic medium where the only source of C and N was asparagine. Of the investigated characteristics the cultivation medium influenced most the adherence of isolates to xylene as after cultivation in a synthetic medium all isolates with the exception of two were hydrophobic. The motility of strains was also greater after growth in this medium. Conversely cultivation in complete medium suppressed these properties of isolates. The cultivation medium did not influence the adherence of isolates to plasts and only a slight influence was observed on the salt aggregating properties of the investigated strains. PMID- 11881301 TI - [Trauma care system in Turkey and the approach to patients suffering head trauma]. PMID- 11881302 TI - [The effects of abdominal compartment syndrome on the serum urea and creatinine levels]. AB - BACKGROUND: The purpose of this study was to assess the effects of abdominal compartment syndrome (ACS) on the kidneys. METHODS: Intra abdominal pressures (IAP) were indirectly measured through urinary bladder. The patients were categorised into four groups according to IAP levels. Serum urea and creatinine levels and IAP were measured once a day. Abdominal decompression was planned according to IAP as well as clinical assessment. RESULTS: The number of patients in this study was 25. Serum urea and creatinine levels were highest in the group IV (group in which abdominal pressure was above 31 cmH2O) (t > 0.05). Five of the 25 patients were died and anuria developed in these five patients before death. In three of five patients abdominal decompression operations were performed. CONCLUSION: ACS is an unusual and often lethal syndrome. The most important treatment is abdominal decompression and we conclude that it should be done in patients with IAP 30 cmH2O or above to protect renal function. PMID- 11881303 TI - [Is jejunal feeding efficient in critically ill patients?]. AB - BACKGROUND: The aim of this study is to compare the results of jejunal and gastric nutrition in the ICU. METHODS: Caloric intake and nutritional complications were recorded for ten days period in patients receiving gastric (n = 21) and jejunal (n = 22) feeding. RESULTS: Caloric requirements were reached on the 3rd day of nutrition in 86% of jejunal and 28% of gastric feeding patients (p 0.001). In jejunal group, delivered calorie/goal calorie ratio was found 15-20% higher than the gastric group. Serum albumin, triglyceride, cholesterol levels and nitrogen balance did not show significant differences between groups. Vomiting (p 0.01) and colouring of tracheal aspirates (p 0.05) were more frequent in gastric group, however positive tracheal culture frequency did not differ between the groups. CONCLUSION: It is concluded that higher caloric intakes could be tolerated earlier in patients receiving jejunal feeding. PMID- 11881304 TI - [Determination of malondialdehyde level in peritoneal fluid in patients with acute abdomen]. AB - BACKGROUND: Our aim is to determine the relationship between free oxygen radicals and tissue destruction in Acute abdomen patients. METHODS: In this study, direct determination of free oxygen radicals is succeeded by measuring the Malondialdehyde level (MDA), which is the last product of lipid peroxidation in the peritoneal fluid of acute abdomen patients. Forty patients are included in the study of whom thirty patients were the study group and ten were taken as the control group. MDA levels of both groups and the differences between daily MDA levels of each group are compared statistically by using one way ANOVA, student t test and student Newman Keuls test respectively. RESULTS: There was statistical significant difference between study and control group (p < 0.05). Also there was significant difference between MDA levels of infected, non-infected and control groups (p < 0.01). It was understood that high MDA levels in the study group source from infected group and there was no significant difference between non infected and control group. CONCLUSIONS: In this study we proved the high MDA levels in the peritoneal fluid of Infected Acute Abdomen patients. This result is meaningful for the diagnose and following of these patients. PMID- 11881305 TI - [Evaluation of percutaneous endoscopic gastrostomy and results]. AB - BACKGROUND: The aim of this study is evaluate the percutaneous endoscopic gastrostomy (PEG) and its complications in order to provide enteral nutrition support for a long time period. METHODS: PEG tube was applied to 15 patients aged between 18-95 years, who could not feed orally prophylactic antibiotic didn't use to any patient. After 8 hours hungry PEG was applied by using pull technique and the results were evaluated prospectively. RESULTS: PEG was successfully done in 14 patients. Gastric juice drainage at the sides of the tube were seen in two patients, and PEG was pulled out in one of them. CONCLUSIONS: PEG is the route of choice for a long lived enteral nutrition, because, it can be applied without general anesthesia, easy to apply, patients can start feeding in a short time period, need shorter hospital stay, can be changed easily, cost effective and has a low rate of mortality and morbidity. PMID- 11881306 TI - [Evaluation of traffic accident cases admitted to the emergency department of the Cumhuriyet University Hospital in 1998]. AB - BACKGROUND: In this study, we aimed to analyze traffic accident cases applying to the emergency department of the hospital of the Cumhuriyet University according to their age, sex, their position during crash, the place and time of the accident, the time to reach hospital, the type of vehicle and their lesion forms. METHODS: The hospital files of cases applying to the emergency department of the hospital of the Cumhuriyet University for traffic accidents, between 01 January 31 December 1998, were investigated retrospectively. RESULTS: Of the cases, most (44.52%) were in the 1-25 age group, 41% was pedestrian. Of the accidents, 81.29% was at the inner part of the city, 44.03% occurred in the summer and 39.83% occurred at 12-18 hours, 44.52% reached to hospital in the first hour. The vehicle type in 60.80% of the cases was automobile, 14.28% of the cases involved in alcohol. The blunt injuries were seen usually, 47.50% of which was present in the head-neck region. 257 cases (41.45%) were hospitalized. 23 cases (3.71%) died because of traffic accidents. CONCLUSIONS: The similar studies in literature were examined. The factors causing the traffic accident and the problems resulting from traffic accident were pointed out. The treatment manner in the traffic accident cases was suggested. PMID- 11881308 TI - [Preliminary results for treatment of tibial plateau fractures with the Ilizarov technique of ligamentotaxis]. AB - BACKGROUND: The results of the eight patients who admitted to emergency room with tibial plateau fractures and treated with Ilizarov technique, were retrospective evaluated both clinically and radiologically. METHODS: Seven (87%) men and one (13%) woman ranging in age from 23 to 38, were evaluated. All the cases were between type IV to VI according to Hohl classification preoperatively. Closed indirect reduction by ligamentotaxis was attempted in all fractures, no open technique was performed. RESULTS: Three (38%) cases had open fractures (type I according to Gustilo-Andersen Classification) preoperatively. Six (75%) traffic accident and two (25%) falling from a height were detected as etiology. The cases, whose average follow up was 14 (6-28) months, were evaluated according to Iowa knee score scale and seven (87%) cases were good and excellent. Both clinical and radiological solid fusion were obtained in all cases after removing the frame. Although all the cases had minimal to moderate pin tract infection, all were resolved with dressing and oral antibiotherapy without removing the wires. CONCLUSIONS: Closed reduction with Ilizarov technique is appropriate for treatment of plateau tibia fractures with minimal morbidity. PMID- 11881307 TI - [The effect of antibiotic therapy on lung pathology in experimental models of sepsis]. AB - AIMS: In this study, the effect of an extra-lung sepsis model on lung histopathology is evaluated. METHODOLOGY: In this study 20 Wistar-Albino rats were used. Following the ether anesthesia laparotomy was done. Caecum was ligated by a silk thread and was perforated by 18 gauge needle. It is squeezed until feces emerged. Abdominal wall is closed. By this method peritoneal sepsis was performed. The rats are divided into two groups (n:10). 0.5 ml of serum physiologic is applied to the control group, and imipenem is applied to the antibiotic group as 15 mg/kg/tid. 48 hours later rats were sacrificed by extreme ether anesthesia. Relaparatomy was done and diaphragm was open. Multiple biopsies were made from the lung. Biopsy materials was cultured and examined histopathologically. RESULTS: In control group; rats died in 48 hours (%100), but antibiotic group were alive (%0). The results of lung biopsy cultures are; in all rats in control group, Escherichia coli (E. coli) and Bacteroides fragilis (B. fragilis) were cultured (%100). Whereas in antibiotic group there is no bacteria cultured (p < 0.001). Histopathologic results are: in control group there was wide spread edema and congestion and inflammatory reaction. In antibiotic group there was slight edema, congestion and inflammatory reaction. CONCLUSION: In septic condition sings of adult respiratory distress syndrome occurs. Large spectrum antibiotics can prevent bacterial translocation in lungs and could minimize the lung injury. PMID- 11881309 TI - [Clinical differences between surgically treated patients with large bowel obstruction in the cities of Van and Istanbul]. AB - BACKGROUNDS AND AIMS: Geographical differences between cases of colonic obstructions affect clinical course and outcome of patients. We aimed to establish regional clinical differences between patients with colonic obstruction from eastern and western regions of Turkey. METHODS: We retrospectively analysed 224 patients with colonic obstruction who were surgically treated in two hospital situated in Istanbul from western and in Van from eastern regions of Turkey. This analysis was made in respect of demographic features, causes of obstruction and location in the colon, complicated obstructions, postoperative clinical course and mortality. RESULTS: Men constituted 71.4% of patients with a mean age of 55.5 years. Obstruction site was the left colon in 82.5% and the sigmoid in 66% of patients from western region, and 91.7% (p = 0.03), and 85% (p = 0.007) respectively of those from eastern region. The leading causes of obstruction were obstructive cancer (52.5%) in Istanbul and volvulus (80.2%) Van respectively (p < 0.001). The colonic obstruction was complicated in 22% of patients. The rate of complicated obstruction was 17.5% and 26.4% (p = 0.07) in Van respectively. The overall postoperative mortality was found as 12.9%. The mortality being 8.6% in simple obstruction raised to 28% (p = 0.008) in complicated patients. CONCLUSIONS: We found significant differences in colonic obstruction cases between eastern and western parts of Turkey. The incidence of complicated obstruction is bigger in eastern region. A considerable mortality arises in surgically treated patients with colonic obstruction. Postoperative mortality was significantly elevated in cases of obstruction complicated by strangulation, necrosis, and perforation. PMID- 11881310 TI - [Intestinal and peritoneal tuberculosis]. AB - BACKGROUND: Abdominal tuberculosis (tbc) is still a medical problem in developing countries. Since it imitates many abdominal diseases, diagnosis can be easily missed unless the disease is suspected. METHODS: The aim of this study to evaluate the value of clinical, physical and laboratory findings and to discuss the diagnostic and therapeutic options in 121 patients with intestinal and peritoneal tbc. The diagnosis was made by histopathological examination of biopsy material and isolation of mycobacterium bacillus in cultures or smears of ascites fluid. RESULTS: The diagnosis was confirmed with laparotomy in 102, laparoscopy in 4, colonoscopy in 6, and percutaneous aspiration in 9 patients. There were intestinal tbc in 67 (55.3%) patients and peritoneal tbc in 54 (44.6%). Intestinal involvement was commonly located at ileocecal area. Anti tuberculous chemotherapy was started and avoided from extensive resection in surgical treatment. There were a total of 87 complications in 52 patients (42.9%) at the postoperative period. Wound infection was the most frequent complication. Overall mortality rate was 13.2%. The mortality rate in emergency operation was 20.5% while 3.4% in elective conditions. There were no morbidity and mortality in patients whose diagnosis were made by conservative procedures. CONCLUSIONS: Laparoscopic endoscopic and percutaneous aspiration procedures are useful for diagnosis in the selected cases of intestinal and peritoneal tbc. Laparotomy should be performed only when complication develops or diagnosis is uncertain. Extensive resection should be avoided in surgical treatment of intestinal tbc. Early diagnosis and treatment will decrease the complications that can be develop during the progress of the disease and consequently the mortality rates. PMID- 11881311 TI - [Importance of source control in secondary peritonitis]. AB - AIMS: In this study we discussed retrospectively secondary peritonitis patients, surgical treatment modalities and their effectiveness in our clinic. MATERIALS & METHODOLOGY: 91 patients were operated due to diffuse peritonitis between December 1998 through July 2001 in our clinic were analysed by age, sex, etiology of peritonitis, treatment modalities, morbidity and mortality. RESULTS: 32 patients were female (35.2%) and 59 patients were male (64.8%). The median age was 40.4. The most common etiologic factor for secondary peritonitis is peptic ulcus perforation (38.4%). 23 cases (25.2%) had the diagnosis only by physical examination and laboratory assessment. The other cases diagnosed with roentgenographic evaluation. In the first operation, the aim was the source control and eradication. In 3 cases, planned re-laparotomy "staged abdominal repair" (STAR) were performed. Conservative treatment modality was performed for 8 cases. The median hospital stay for the patients were 7.4 day. Morbidity was encountered in 13 (13.6%) and mortality was encountered in 2 (2.1%). CONCLUSION: Source control must be the primary aim of the first operation for secondary peritonitis patients. If there is an uncertainty for source control, STAR procedure should be the choice of the treatment modality for decrease morbidity and mortality. PMID- 11881312 TI - [Thoracic aortic transection following blunt trauma: two case reports]. AB - Acute aortic transection after blunt trauma is a clinical situation, with a high mortality. When the other system injuries are predominant, the diagnosis could be possible only if it would be considered. These type of cases are usually die. The survivors are generally the cases of whom be diagnosed incidentally. Two cases who were admitted to our clinic due to falling down (suicide attempt) were diagnosed as aortic transection and were immediately operated. End to end graft interpositions were performed by using left atrio-femoral bypass in one patient and femoro-femoral bypass in the other. Systemic examinations of all the patients who are referred to the hospital after blunt trauma should be done completely and the physician should be alert for possible aortic transection or dissection. PMID- 11881313 TI - [Pregnancy-related, rarely-seen spontaneous lumbar artery rupture]. AB - Some arterial aneurysm ruptures can be seen during pregnancy. However, spontaneous rupture of the lumbar artery in pregnancy are very rare. In that manuscript we present a 22 year old female who is 8 months pregnant, seen at a peripheral hospital for suspicion of placenta decolmant. When the fetal heart sounds were missed, the patient was transported to our hospital. We reoperated on her as an emergency. We found that a spontaneous rupture of the lumbar artery. PMID- 11881314 TI - Posttraumatic distal radioulnar synostosis and distal radial epiphyseal arrest. PMID- 11881316 TI - [Replantation in the complete amputation of the upper extremity]. AB - Trauma of the upper extremity can be a cause of significant morbidity and disability to otherwise productive people. Wounded extremity can be saved by replantation of the amputated parts. Replantation of traumatic amputations can be performed with reasonable success at a regional medical center when experienced surgeons, appropriate and experienced equipment, and skilled ancillary care are available. Successful replantation significantly reduces the morbidity of upper extremity amputations. We presented one case that, 9 year-old, was bridged to our hospital because of total arm amputation after sustained during use of a lawn mover. His arm was saved by urgency surgery. PMID- 11881315 TI - [The metabolic and hormonal effects of perioperative ibuprofen treatment on surgical stress]. AB - BACKGROUND: Surgical stress induces hormonal and cytokine responses proportional to the extent of the injury. Perioperative administration of cyclo-oxygenase inhibitors reduces cytokine production and nitrogen losses. The objective of this study is to evaluate clinically the metabolic and hormonal effects of ibuprofen which is cyclo-oxygenase inhibitor on surgical stress. METHODS: A prospective clinic study was performed in 20 patients who undergone thyroidectomy. Patients were randomly divided two groups as ibuprofen group (n = 10) and control group (n = 10). In the ibuprofen group, pills containing ibuprofen (400 mg) were administered orally 12 and 2 hours before surgery, and every 8 hours until the third postoperative day. In the both groups, blood samples were collected 24 and 2 hours before surgery and 1, 4, 6, 24, 48, and 72 hours after skin incision for glucose, CRP, leukocytes, ACTH, cortisol determinations. RESULTS: When preoperative values were compared with postoperative values, surgical stress caused significantly changes on the all parameters in the both groups. The highest levels of ACTH and cortisol were at 1st and 6th hours respectively. In both groups, plasma cortisol levels remained elevated for 3 days, whereas plasma ACTH levels returned to the basal level at 1 day. In the ibuprofen group, the levels of ACTH and cortisol were significantly less than those of control group (p < 0.001 and p < 0.001). In the ibuprofen group glucose level (p < 0.001) and count leukocyte (p < 0.001) increased mildly. The level of CRP increased gradually after first day and were high until 3rd days (p < 0.01). In the ibuprofen group the fever increased smaller (p < 0.01). CONCLUSIONS: In conclusion, in perioperative period pretreatment with ibuprofen is able to reduce the neuroendocrine and humoral responses, but it can not completely suppress. Our present study suggests that ibuprofen may be useful in decreasing the surgical stress response in the serious patients. PMID- 11881317 TI - Anomeric specificity of glucose metabolism in health and disease. PMID- 11881318 TI - Hepatic fibrin-ring granulomas in granulomatous hepatitis: report of four cases and review of the literature. AB - BACKGROUND: The differential diagnosis of hepatic fibrin-ring granulomas includes infective agents (Coxiella burneti, CMV, EBV,....), hypersensitivity to medication (allopurinol) and malignancy. METHODS: During a period of 6 months, four patients presented at our university hospital with a similar clinical picture of fever and abnormal liver tests, and fibrin-ring granulomas on liver biopsy. Clinical course, laboratory and imaging findings, and histopathological features were compared. RESULTS: Clinical manifestations, and laboratory and imaging findings were similar. Histopathological assessment of the hepatic fibrin ring granulomas appeared not to be helpful in identifying the causative agent. Other histopathological features (e.g. sinusoidal rows of lymphocytes, eosinophilic polymorphonuclear infiltrate) were suggestive for the causative agent, yet conclusive identification was obtained by either serology (Q fever, CMV, EBV), or by exclusion with concomitant stop of medication (allopurinol). CONCLUSIONS: In the differential diagnosis of hepatic fibrin-ring granulomas, serologic titers remain the determining factor, since an infective agent is the most common cause. When hepatic fibrin-ring granulomas are present, other histopathological features may be helpful in making the differential diagnosis. PMID- 11881319 TI - In vitro activity of the new ketolide telithromycin and other antibiotics against Streptococcus pneumoniae in Belgium. AB - In Belgium more than 17% of the invasive pneumococci are not susceptible to penicillin, and more than 38% not to macrolides. The most prevalent mechanism of macrolide resistance in Europe is modification of the drug target site leading to cross-resistance to lincosamides and group B streptogramines (MLSB resistance). Telithromycin is the first antibiotic of the family of ketolides, which differ from erythromycin by having a 3-keto group instead of the neutral sugar L cladinose. We tested the susceptibility of 637 pneumococci, recently isolated from patients in Belgium, to telithromycin and five other antibiotics. Data generated by this study show that telithromycin inhibits 98.4% of pneumococci at a breakpoint concentration of 1 mg/L in spite of a high percentage (> 30%) of strains with the MLSB constitutive type of resistance. Susceptibilities to the five comparator drugs were: penicillin (81.8%), tetracycline (67.0%), levofloxacin (98.9%), erythromycin (61.5%) and clindamycin (66.6%). Consequently telithromycin looks to have considerable potential for the empiric treatment of community-acquired respiratory tract infections. PMID- 11881320 TI - Comparative in vitro activity of temocillin and other antimicrobial agents against Enterobacteriaceae isolated from patients admitted to five Belgian hospitals. AB - Temocillin, a methoxy-derivative of the broad-spectrum penicillin, ticarcillin, has been introduced into clinical practice in Belgium in 1988. Since then, not many surveys of its in vitro activity have been published. This study addresses this issue in a prospective collection of 300 consecutive Gram-negative isolates originating from in-patients in five general hospitals throughout Belgium. In addition to temocillin, seven common antibiotics were tested: amoxicillin clavulanate, piperacillin-tazobactam, cefotaxime, aztreonam, meropenem, ciprofloxacin and amikacin. Meropenem appeared to exhibit the best activity overall, whereas amoxicillin-clavulanate scored the worst. Cumulative MIC plot for two subsets of organisms are given: temocillin, meropenem and cefotaxime are the most active on E. coli and Klebsiella spp., while a significant percentage is resistant to ciprofloxacin and amoxicillin-clavulanate. In the group of inducible Enterobacteriaceae, temocillin, meropenem and amikacin are the most active drugs, while the activity of amoxicillin-clavulanate, piperacillin-tazobactam, cefotaxime and ciprofloxacin is largely decreased. Taking this well preserved in vitro activity of temocillin into account, and looking at its convenient pharmacokinetics and low cost of acquisition, this drug may prove a useful alternative in the treatment of severe nosocomial infections. PMID- 11881321 TI - Hyperoxaluria: an underestimated cause of rapidly progressive renal failure. AB - Hyperoxaluria is rarely considered as a cause of rapidly progressive renal failure. A case is reported of a patient in whom rapidly renal failure developed after subtotal small bowel resection. A diagnosis of calcium oxalate deposits nephropathy was confirmed by renal biopsy. This cause of renal failure may be underestimated and should be systematically searched for in all patients with malabsorption. PMID- 11881322 TI - Hepatotoxicity related to itraconazole: report of three cases. AB - BACKGROUND: The antimycotic synthetic azole compounds are known to lead to toxic liver injury. The occurrence of acute hepatitis is best known for ketoconazole. With itraconazole, hepatotoxic reactions have only very rarely been reported, and histologic data are lacking. We report on three patients who developed acute liver damage during therapy with itraconazole, and in whom liver biopsy specimens were obtained. METHODS: Three patients with apparent itraconazole-induced liver injury were studied. Clinical, laboratory, serologic, and histologic data of all three cases were analyzed. RESULTS: All three patients developed a biochemical histologic pattern of cholestatic liver injury with damage to the interlobular bile ducts. Beginning ductopenia was present in two, suggesting that itraconazole might be responsible for the occurrence of prolonged drug-induced cholangiopathy. Jaundice was the presenting symptom in all three. It was not accompanied by clinical hallmarks of hypersensitivity, which is suggestive for metabolic rather than for immunoallergic idiosyncrasy. CONCLUSIONS: Itraconazole-induced liver injury presents with a cholestatic pattern of injury with damage to the interlobular bile ducts, possibly leading to ductopenia. We suggest that itraconazole should be added to the list of drugs that may be responsible for a drug-induced vanishing bile duct syndrome. Further histologic documentation in other cases is necessary to strengthen our current findings. PMID- 11881323 TI - Giant cell (temporal) arteritis diagnosed following upper limb claudication. AB - Giant cell arteritis or temporal arteritis occurs almost exclusively in people over 50 years of age. It classically presents with new onset temporal headache, scalp tenderness and jaw claudication. Proximal muscle pain and stiffness is often present because of frequent association with polymyalgia rheumatica. In most cases, the erythrocyte sedimentation rate is markedly elevated. Uncommon presentations include systemic symptoms and symptoms related to large artery involvement. We report a case of giant cell arteritis without symptoms related to the temporal artery, diagnosed angiographically following upper limb claudication and confirmed by temporal artery biopsy. PMID- 11881324 TI - Usefulness of [99mTC]MIBI and [18F]fluorodeoxyglucose for imaging recurrent medullary thyroid cancer and hyperparathyroidism in MEN 2a syndrome. AB - We report the case of a MEN 2a patient with a history of medullary thyroid cancer (MTC) treated by total thyroidectomy, who presented an increasing calcitonin level, suggesting tumor recurrence. Conventional radiographic and radionuclide imaging failed to localize the responsible lesions. A planar and tomographic (SPECT) [99mTc]MIBI scan, performed in order to investigate a recent hyperparathyroidism localized a parathyroid adenoma and revealed an abnormal uptake in the left lateral neck region, corresponding to apparently banal lymph nodes on MRI. This abnormal uptake was also observed on a [18F]fluorodeoxyglucose positron emission tomography (FDG-PET) study and was proven to be an uptake in MTC lymph nodes metastases as confirmed by histopathologic analysis. We conclude that, using an adequate acquisition protocol (i.e. SPECT), [99mTc]MIBI scan is potentially able to localize both parathyroid adenoma and recurrent MTC at one and the same time, particularly in case of non-diagnostic conventional imaging techniques. In this setting, the potential usefulness of FDG-PET is also discussed. PMID- 11881325 TI - Reticulocyte enumeration on Bayer Advia 120 is not influenced by malaria parasites. PMID- 11881326 TI - [Preimplantation diagnosis and therapeutic clones]. PMID- 11881328 TI - [Medical treatment of fever and recurrent convulsive attacks]. PMID- 11881330 TI - [Diets for weight reduction]. PMID- 11881329 TI - [Vasoprotection through vascular nitric oxide. Molecular basis of antiatherosclerotic actions]. PMID- 11881331 TI - [Neural stem cells in the brain]. AB - Stem cells in the central nervous system were usually considered as relevant for evaluation only in embryonic time. Recent advances in molecular cloning and immunological identification of the different cell types prove the presence of neurogenesis of the new neurons in adult mammals brains. New neurons are born in two areas of the mammal and human brain--sybventricular zone and subgranular zone of dentate gyrus. New born granular neurons of dentate gyrus have a great importance for memory and learning. New neurons originate from precursors which in culture and in situ could also transform into astrocytes and oligodendrocytes, thus fulfill criteria of neural stem cells. In culture, mitotic activity of these stem sells depends on fibroblast growth factor 2 and epidermal growth factor. Depletion of cultural medium of these factors and addition of serum, other growth factors (Platelet-derived growth factor and ciliary neurotrophic factor) leads to generation of neurons and astrocytes. Isolation and clonal analysis of stem cells is based on immunological markers such as nestin, beta-tubulin III, some types of membrane glicoproteids. Identification and visualization of stem cells in brain revealed two populations of cells which have properties of stem cells. In embryonic time, radial glia cells could give origin to neurons, in mature brain cells expressing glial fibrillar acidic protein typical marker of astrocytes fulfill criteria for stem cells. Neural stem cells could transform not only into mature neurons and glial cells but also into blood cells, thus revealing broad spectrum of progenitors from different embryonic tissues. Further progress in this field of neurobiology could give prosperity in the cell therapy of many brain diseases. PMID- 11881332 TI - [Neuropeptide thyroliberin--an endogenous anticonvulsant in the brain]. AB - Thyroliberin (TRH) promoting endogenous antidepressive effect is the most general regulator of the central mechanisms and visceral functions (especially respiration). Our group pioneered in applying the anticonvulsant action of TRH after local intranasal application. This application TRH in ultra-low doses contrast the method of systemic TRH administration (i.v., i.m. or oral in the large doses--mg). In our experiments intranasal application of 10(-9) M, 10(-10) M and 10(-12) M TRH significantly inhibited the severe epileptic motor fits in rats induced by pentylenetetrazole (PTZ). Beneficent effect of TRH is also confirmed by EEG (TRH suppressed SWD in cortex, amygdala and hippocamp). In the experiment that follows compared effects of TRH (pyroGlu-His-Pro-NH2) and its metabolite dipeptide cHis-Pro-NH2 (10(-10) M, 10(-5) M). The experiments make more precise that only TRH but not His-Pro posses the anticonvulsant properties. There is a good believe that medical potentialities of TRH have not been exhausted and its new possibilities of its usage will be revealed in epileptology. PMID- 11881333 TI - [Functional organization of the vestibulospinal system in amphibians]. AB - In experiments on the preparation of a frog perfused brain, using recording of intracellular potentials the vestibulospinal neurons were identified on the basis of excitatory postsynaptic potentials evoked by the stimulation of the ipsilateral vestibular nerve and antidromic activation from the stimulation of the cervical and lumbar enlargements of the spinal cord. The average conduction velocity determined for axons of C neurons was 10.67 m/s and for L neurons 15.84 m/s. The ratio of C and L neurons over the vestibular nuclear complex was very stimular to each other: 52% C neurons and 48% L neurons. The majority of both types of neurons were localized in the lateral vestibular nucleus (58.6%), to the lesser extent in the descending vestibular nucleus (30.7%) and very little in the medial vestibular nucleus (10.6%). Fast and slow cells were detected among the vestibulospinal neurons. The fast neurons of L cells did not prevail greatly over the slow ones, whereas the slow neurons of C cells prevailed comparatively largely over the fast neurons. Thus, it became possible to reconstruct spatial distribution of the identified vestibulospinal neurons. The results of spatial distribution of C and L vestibulospinal neurons in the frogs failed to conform to definite somatotopy, which is characteristic for mammalian vestibular nuclei. C and L neurons in the frog's vestibular nuclei as a source of vestibulospinal fibres, are scattered separately or more frequently in groups, so that they establish a "patch-like" somatotopy and do not form a distinctly designed fields as in mammals. PMID- 11881334 TI - [A unified postsynaptic mechanism for the effect of various neuromodulators on modification of potentiated and depressed inputs to hippocampal cells (hypothesis)]. AB - The unitary postsynaptic mechanism underlying the influence of diverse neuromodulators on modification of excitatory and inhibitory inputs to granule, pyramidal and inhibitory hippocampal cells is suggested. According to this mechanism, the effect of dopamine, adenosine, acetylcholine, noradrenaline, serotonin, somatostatin, galanin, opioids, cannabinoids, neuropeptide Y on postsynaptic receptors, bound to Gi/0 proteins, should promote LTD of excitatory inputs and LTP of inhibitory inputs. The effect of dopamine, adenosine, acetylcholine, noradrenaline, serotonin, vasopressin, tachykinin, histamine on postsynaptic receptors, bound to Gs and Gq/11 proteins, should oppositively modulate the same inputs. Only synaptically activated excitatory and inhibitory inputs can by influenced by neuromodulators. The character of neuromodulatory influence on modification of hippocampal synaptic efficacy, implying from the suggested mechanism is in accordance with known experimental data. PMID- 11881335 TI - [Interlevel relations in neural memory: extrasynaptic reception of mediators, potentiation, and spontaneous activity]. AB - The firing of "spontaneous" spikes is regarded as a result of mediator propagation to extrasynaptic receptors. Receptor-receptor interaction unites them in dimer and dimer clusters, which accept three conformational states under agonist action. There are two cooperative and potential dependent transitions between the states, where cluster accumulates or releases energy. The released energy can trigger a mechanism of endogenous (spontaneous) neuron firing in potentiation condition. These accumulating and triggering properties are absent in third (passive) conformational state, where gating charges immobilization reduces conformational mobility. The features of ionotropic, metabotropic and combined mediator action are discussed for different level of slow potential. Conformational effect depends on conformity of pattern space-temporal structure to geometric and functional features of metabotropic mediator sources in cluster environment. Each cluster appears to be adjusted for recognizing a certain vast set of afferent patterns. Number, structure and dimensionality of the recognized patterns are given by: 1) threshold of conformational transition, 2) allocation of synaptic and extrasynaptic mediator ejecting points in gap-hole environment of the receptive cluster 3) combinatorial connections of presynaptic cells with inhibit and excite synapses and 4) signal delays in presynaptic ways and neuropil. Numerous receptive clusters of soma-dendrite membrane are capable to write down information, to keep and accumulate it and to recover. Engram stored as passive/active conformational receptive cluster states is recovered in inversion by "spontaneous" neuronal activity. The original information may be recovered by reading via inhibit synapses. PMID- 11881336 TI - [Homeostasis of the gastric mucosa and blood circulation. 2. Role of ischemia in disruption of the gastric mucosa]. AB - To date there is a lot of data of literature indicating that microcirculatory disorders play the main role in the development of gastric mucosal damages induced by stress, ethanol, nonsteroidal antiinflammatory drugs and tobacco smoke. Under stress gastric mucosal blood flow disorders may be caused by the actication of sympathetic nervous system. Ulcer healing is accompanied by the angiogenesis and by the increase of blood flow in the ulcer border and tissues surrounding the ulcer. Therefore now the main studies are concentrated on the search of defence-enhancing agent rather than drugs for antisecretory therapy. Therapeutic strategy suggests the use of some potential vasodilators such as NO donors, prostaglandin analogues, oxygen radical scavengers, endothelin, leukotrienes, platelet-activating factor antagonists and/or their synthesis inhibitors. At present, the therapeutic possibilities seem to be restricted and nothing indicates that stimulation of the defensive factor only, is more effective in the treatment of peptic ulcer than inhibition of aggressive factors. However we suggest that blood flow correction may be very important for the treatment of refractory ulcers or for prophylaxis of stress ulceration and peptic ulcer recurrence. PMID- 11881337 TI - [Hyperinsulinemia as a factor preceding body mass increase in liquidators of the Chernobyl nuclear accident]. AB - Basal hyperinsulinemia has been recordable over several years in liquidators of the effects of Chernobyl Accident (ChAEL). There was no dependence of hyperinsulinemia on age, body mass or lenght of service of those persons having worked within the 30-km zone. The findings secured suggest to us that increase in the body mass index (BMI) in ChAEL is preceded by a rise in the blood level of insulin; there is no relatedness of the basal level of insulin in the blood to BMI. PMID- 11881338 TI - [Aspects of regional hemodynamics in stress-induced fractures and pathological reorganization of shin bones]. AB - In the examination of 58 servicemen presenting with a pathological reorganization and stress-induced fractures of shin bones, disturbances have been revealed in the limb regional hemodynamics manifested by an increased artery tonicity and peripheral resistance, a decreased arteriole, venule tonicity and an excess of blood flowing through them, a slowed-down bloodflow, disordered microcirculation, in the wake of which events the plasma passes into the intertissue spaces, there develops lymphostasis and edema of soft tissues. The results of the studies carried out permit making an optimum choice of pathogenetically validated treatment options from among those designed to deal with the above troubles. PMID- 11881339 TI - [Efficiency of sorption therapy with the enterosorbent BC-1 in experimental hypercholesterolemia]. AB - A hypercholesterolemic effect has been examined of the enterosorbent BC-1 capable of binding cholesterol from the standardized solution and bile in in vitro experiments in rats with induced hypercholesterolemia. The results secured suggest to us a high sorption capability of the new sorbent with respect to cholesterol. PMID- 11881340 TI - [Effects of maternal toxic hepatitis on the postnatal development of duodenal morphology of their offspring]. AB - Retardation has been shown in development and formation of the vascular-and tissue structures of white rat younglings (3, 7, 14, 21, 30 days old) born to mothers with chronic heliotrine hepatitis in the duodenum versus control animals. In the duodenal mucosa, the villi are short, their shape lost, the mucosal muscular plate development returded. Noted in the epithelium are distrophic changes, a decline in the epithelial cell mitotic activity. The intraorgan blood vessels are dilated and blood-filled. There are spots of extravasation. PMID- 11881341 TI - [Experimental studies on the effect of ceramic materials on cellular and humoral immunity]. AB - It has been established in an experimental setting in laboratory animals through testing a number of reactions such as active skin anaphylaxis reaction, mast cell degranulation reaction, specific leucocyte lysis reaction, delayed hypersensitivity reaction, and graft-versus-host reaction that ceramic preparations hydroxilapatite M and osteogel-7 have no sensitizing effects; osteogel-7 is not endowed with immunomodulating activity, which fact suggests to us its immunological inertness. PMID- 11881342 TI - [Evaluation of quantum therapy for the treatment of stable angina]. AB - A total of 68 patients have been examined. Of these, 21 had functional class (FC) I stabile angina, 23 presented with FC II angina, 24 had FC III angina. Instituted in the control group patients (n = 30) was standard antianginal therapy (SAT). Laser therapy against the background of SAT employed has been found to improve the functional state of the myocardium, enhance tolerance to physical loads, improve indices for intracardiac hemodynamics. He-Ne laser irradiation has an analgecizing effect. Patients with FC I-III exertional angina can derive benefit from laser therapy due to its cardioprotective effect. Positive hemodynamic shifts were accompanied by improvement in general health of patients manifested by lower frequency of angina attacks and episodes of pain free ischemia of the myocardium. Laser therapy had an effect on relation between painful and painless ischemia of the myocardium as evidenced by a predominant decrease in pain-free episodes of myocardial ischemia, this being regarded as a prognostically favourable fact. PMID- 11881343 TI - [Assessment of treatment efficacy of essential hypertension using lateral therapy techniques]. AB - As many as 58 patients with stage II essential hypertension were studied for their brain effects of lateral colourlight therapy and zonal-lateral electrical stimulations. Straight light, reverse light and electrical stimulations have been found to be associated with an instant antihypertensive effect, while electrical stimulations had a hypertensive effect. The author has come to the conclusion that the studied methods of lateral therapy of reverse polarity can be used as treatment of hypertensive crises. PMID- 11881344 TI - [Computerized acupuncture methods in the diagnosis of autonomic nervous system disturbances]. AB - Described in the paper are methods of electropuncture diagnosis (Nakatani methods, auricular diagnosis, Foll's method). An algorithm of examination with making use of the above methods is suggested. A combination is validated of methods of electropuncture diagnosis in a comprehensive evaluation of those patients presenting with affections of the nervous system. PMID- 11881345 TI - [Combination phytotherapy in the treatment of heart failure]. AB - In the article, data from the published literature are summarized together with results of the author's investigations relevant to aspects of pharmacotherapeutical efficacy of cardiac glycosides, plants endowed with antihypertensive and diuretic activities in acute and chronic heart failure. Pharmacological mechanisms of action of a new combined vegetal drug, chomvikorin N, are described in detail; its clinical efficacy is shown in the treatment of heart failure. PMID- 11881346 TI - [Use of extracts of Glycerrhiza glabra L. in the correction of some indices of local nonspecific defense in patients with protracted pulmonary pneumonia]. AB - Results are presented of studies on local nonspecific defence in patients with protracted pneumonia in the bronchoalveolar lavage fluid (BALF) during the course of conventional therapy involving the use of T-activin and extractum Glycerrhiza glabra L. The course of conventional therapy has not been shown to be associated with a substantial normalization of cytosis in the BALF cell precipitate or augmentation of sIgA content of lysozyme. T-activin makes for a reduction in the content of mature neutrophilous granulocytes and for an increase in humoral factors of defence. Extractum Glycerrhiza glabra L. has been found to be superior to T-activin in diminishing neutrophilic granulocytes count, increase in the BALF content of macrophages, lysozyme, s IgA; it proved to be endowed with an antiphospholipase activity, which facts predetermine apperant efficacy of the drug with respect to the lung local defence system in those patients presenting with protracted pneumonia. PMID- 11881347 TI - [Isoptin in the therapy of supraventricular arrhythmia in patients with diabetes mellitus, non-insulin dependent, concurrent with ischemic heart disease]. AB - The article focuses on the action of isoptin as a monotherapy and in the complex with mildronat in 67 patients with non-insulindependent diabetes mellitus (NIDM) and supraventricular arrhythmia. Isoptin has been shown to be a very effective drug in patients with atrial disorders of cardiac rhythm suffering from diabetes mellitus and ischemic heart disease, especially so in a complex therapy with mildronat. The results obtained permit recommending isoptin as an antiarrhythmic drug in NIDM patients. PMID- 11881348 TI - [Effect of capoten on central hemodynamic indices in children with different clinical forms of chronic glomerulonephritis]. AB - The paper contains a summary of results of a comparative analysis of efficacy of effects of the orthodox and combined therapy on parameters associated with the central hemodynamics. It is shown that conventional therapy supplemented with antihypertensive remedies makes for a more prominent effect concerning stabilization of hemodynamic indexes on the central level. This is particularly important in those children presenting with the nephrotic form of the disease. PMID- 11881349 TI - [Structural and functional status of the hypophyseal-thyroid system in prenatally irradiated children]. AB - In order to evaluate the morphologic-and-functional state of the hypophysis thyroid system long after the Chernobyl accident we examined 1491 children from the northern territories of the Zhitomir region. Of these, 261 had not been in utero exposed to radioiodine, 1230 pediatric subjects proved to be postconception exposed. In utero radioiodine has not been found to affect the thyroid size in any noticeable way. The degree of structural-and-functional indices for the thyreostat system in prenatally irradiated children depends on the stage of the thyroid development just when there happened to be an exposure to radioiodine. PMID- 11881350 TI - [Experience gained with nimodipine (nemotane) in the treatment of patients with cerebral hemorrhage]. AB - A clinical and paraclinical monitoring was conducted of the condition of those patients presenting with cerebral hemorrhage using X-ray computerized tomography, ultrasound dopplerography of extra- and intracranial arteries. Shown in the study is a positive effect of nimodipine on the risk of development of clinically significant neurological deficit with no regression undergone against the background of paraclinically recordable objective signs of constrictive-stenotic artheriopathy and formation of unfavourable (gross invalidism, vegetative state, case mortality) outcome according to the Glasgow Outcome Scale and low level of functional activity according to the Barthel Index at day 30 after the development of hemorrhage. The drug can be recommended for use in the treatment of aneurysmatic cerebral hemorrhages. PMID- 11881351 TI - [Effect of Erbisol on indices of endogenous intoxication in patients with microbial eczema]. AB - Identified in 66 patients with microbial eczema were changes in the blood serum and urine levels of medium-weight molecules. In a combined treatment of the test group patients the home-produces drug erbisol has been shown to secure good results, which fact permits recommending the use of erbisol in the therapy of microbial eczema. PMID- 11881352 TI - [Efficacy of ceruloplasmin in the resuscitation oncology clinic]. AB - The use of ceruloplasmin in the intensive therapy of oncological patients has been shown to optimize the course of the rehabilitative period promoting dissipating of polyorgan failure, restoration of oxidant-antioxidant balance, reduction in the incidence of pyo-septic complications. Ceruloplasmin is found out to have an apparent detoxicating and antiinflammatory effect, it activities the bodily antioxidant defence, favors normalization of the condition of the immunity lymphocytic link. PMID- 11881353 TI - [Immunological variants and free radical characteristics of workers exposed to epoxy resins]. AB - Workers on exposure to epoxy resins (ER) have been shown to reveal several variants of immunogrammes of blood, expression of Fas-ligands and oxygen dependent metabolism of phagocytes. These are as follows: the immune deficiency variant (38.1%), the autoimmune one (33.3%), the state of norm (28.6%). The blood phagocyte oxygen-dependent metabolism tends towards activation but the phagocyte functional reserve is reduced which facts suggest depression of metabolism and phagocytic defence in the organism. Identified in the study are several variants of kinetic parameters of the blood serum chemiluminescence suggesting activation/inhibition of process of free radical oxidation, strengthening/weakening of the blood antioxidant defence, which events can be regarded as early diagnostic and prognostic criteria of adverse effects of epoxy resins on the organism of those handling them. These data suggest to us an insufficient reliability of the existing maximum permissible concentration of epoxy resins in the air of the working zone and attest to the need for its reconsideration. PMID- 11881354 TI - [Ways to improve the health of the population of Ukraine]. AB - An analysis was performed of the health status of the population of Ukraine with reference to social-and-economic, ecological, and medical factors affecting it. Especial emphasis is placed on prophylaxis within the whole health system. It is pointed out that positive results are to be achieved through pursuing the scientifically substantiated policy of administering prophylaxis. Such a policy is deemed warranted at both regional and national levels. It is emphasized that the existing health system of Ukraine requires adaptation under present-day conditions in the state, financing of certain programmes to ensure improvement of health in the best way possible. PMID- 11881356 TI - [System of collection and dynamic processing of information for predicting population genetics]. AB - Creation has come to be necessary in Ukraine of a system for collection, dynamic supply and processing of information about genetical processes. Such a system as that described above is submitted by the authors. The basis of the system is a module in which, by using special units and connections between them, a collection, storing, and processing of information about genetic pathological processes in the time-related course is to be carried on together with prognostication thereof in the population of the country as a whole or in each region taken separately. PMID- 11881355 TI - [Complex characterization of the quality of hospital care provided patients with glomerulonephritis (as per a regional nephrology department)]. AB - Based on expert examination of 296 histories of 145 patients with glomerulonephritis the frequency of errors being made in examining of patients at the prehospital stage, in hospital, has been estimated. Kinds of errors include diagnostic errors (primarily in defining forms and variants of the disease), errors in administering treatments to patients. This necessitated raising the level of the primary link in the nephrological educational program for medical men and warrants development of alternative forms of the in-patient medical care delivery to patients with glomerulonephritis. PMID- 11881357 TI - [Role of the pulmonary surfactant in the pathogenesis of bronchial asthma]. AB - The submitted data from the published literature coupled with findings from the authors' own investigations underline the importance of the part that disturbances in the system of the pulmonary surfactant have in the pathogenesis of bronchial asthma. This permits regarding the substitution surfactant therapy with exogenous surfactants as a pathogenetically validated treatment modality for bronchial asthma as evidenced by the first positive results of its employment. PMID- 11881358 TI - [Role of environmental factors in patients with circulatory disturbance syndrome who developed low-dose radiation-induced peptic ulcer]. AB - 98 patients actually in the zone of abnormal radiation developed peptic ulcer with its characteristic clinical manifestations as evidenced by a morphological study of the mucus and values for gastric secretion. The findings secured are considered to be a manifestation of parasympaticotonia-related discirculatory syndrome, which fact warrants institution of a corrective therapy. PMID- 11881359 TI - [Analysis of causes of peptic ulcer recurrence in patients in Kiev]. AB - The paper presents an analysis of causes of relapse of gastric and duodenal peptic ulcers based on review of 2120 case records of patients from the Ukrainean German gastroenterological centre over the period six months--two years. Chief causes of relapse of gastric and duodenal peptic ulcer are identified, such as incomplete course of eradication therapy; failure on the part of the patients to keep to the physician's orders, which fact suggests to us that there has not been achieved the necessary "compliance"; resistance to the therapy treatments administered, in the first place, to metronidazole (36%), development of gastroestrophagitis reflux. PMID- 11881360 TI - [Cytoprotective properties of smecta on the intestinal mucosa of the alimentary canal]. AB - The chief factors of aggression are discussed together with mechanisms of protection of gastric and duodenal mucosa. Results of treatment of 30 patients with gastritis (Helicobacter pylori negative) with a cytoprotective drug smecta are presented. PMID- 11881361 TI - [Differential diagnosis in "undifferentiated" inflammatory diseases and dyskinesias of the large intestine]. AB - The majority of foreign gastroenterologists have their doubts whether there exist "undifferentiated" colitisis. They consider these to be a variant of the irritable bowel syndrome. We have examined 173 patients. Of these, 67 percent were less than 45 years of age with significant intestinal disorders caused by chronic colitis. Diagnosis of Crohn's disease and nonspecific ulcerative colitis had been excluded. The main complaints of the patients were constipations alternated sometimes with short-time diarrhea (61%), diarrhea (13%), spasmodic pains in the inferior parts of the abdomen (20%), abdominal distention, and rumbling (74%). In 79% there was tenderness in the sygmoid colon and caecum and spasm thereof. Rectoromanoscopy (RRS) revealed normal mucosa of the rectum and sygmoid colon in 70%, proctosygmoiditis (predominantly catarrhal) in 30 percent of cases. In proctosygmoiditis (vs the normal mucosa) motor dysfunctions, disorders of the contrast mass passage through the intestine, gaustrations were more often seen but the relief of the mucosa was intact. Results of morphological investigations of biopsy specimens of rectal and sygmoidal mucosas were compared with endoscopy findings. No histological changes were revealed in 23% patients with proctosygmoiditis whereas in cases of endoscopically normal mucosa moderate inflammation was rarely detectable. Inflammatory changes in the mucosa were to be seen more frequently by cytologic analysis than by biopsy but more rarely than by RRS. In this way, 25% patients with endoscopical diagnosis of proctosygmoiditis had normal cytological picture. The increased amount of intestinal enzymes (enterokynase, alkaline phosphatase) in stools does not permit differentiating the functional and organic types of the disease, but it significantly contributes to endoscopical, morphological, cytological, and clinical data. Our investigations show that "undifferentiated" colitis is a rarity and that one finds difficulty in differentiating between the above condition and the irritable bowel syndrome. PMID- 11881362 TI - [Current approaches to the diagnosis and treatment of irritable bowel syndrome]. AB - Modern definition, classifications, diagnosis and treatment of irritable bowel syndrome (IBS) are presented together with results of treatment of 30 patients with irritable bowel syndrome with the new selective calcium-channel blocker dicetel. PMID- 11881363 TI - [Effect of food deprivation on the proteolytic system in patients with atopic bronchial asthma concurrent with gastroduodenitis]. AB - The article contains data on the state of the proteolysis system in the systemic blood flow and locally in the duodenal mucosa in those patients with bronchial asthma concurrent with gastroduodenitis. Results of fasting dietetic management versus medicamentous treatments showed a complete normalization of the protease antiprotease imbalance with food deprivation. PMID- 11881364 TI - [Effect of anticholinergic therapy on myocardial reserve dynamics in patients with chronic obstructive bronchitis]. AB - The aim of the work is to study the influence of a long-term ipratropium bromide on the character of the functional heart indices changes in patients with chronic obstructive bronchitis. Positive effects on the myocardial reserve are reported under a 2-week atrovent monotherapy. Atrovent favoured a positive trend in clinical symptoms, that determined the myocardial reserve. With the above drug, the frequency of the reduced myocardial reserve has come to be lowered by 34.5%. Based on the data presented recommendations are given for use of ipratropium bromide in chronic obstructive bronchitis patients with the incipient form of heart failure. PMID- 11881365 TI - [Assessment of quality of life in patients with chronic obstructive bronchitis]. AB - The investigation performed in 117 patients with chronic obstructive bronchitis (COB) revealed a significant decline in quality of life (QL) with progressive disease. Decrement of QL level in patients with stage I and II COB has been found to be primarily associated with activation of the sympathetic nervous system and progressing of metabolic myocardial dystrophy. With stage III COB decline in QL is also caused by reduction of volume of the functional pulmonary parenchyma. PMID- 11881366 TI - [Study of the dynamics of immunograms of patients with chronic obstructive bronchitis against the background of treatment with bronchomunal]. AB - The article searches into the efficiency of use of bronchomunal as treatment of chronic obstructive bronchitis (COB). A three-month course of treatment with the drug in 18 COB patients has been found to be associated with a positive dynamics of the most important indices of the immunogram, which fact warrants its administration as an effective COB complications-preventing alternative. PMID- 11881367 TI - [Changes in the blood concentrations of interleukins and electrolytes in miners working in deep coal mines]. AB - Miners working in deep coal mines, engaged in hard physical work under most harsh mine conditions demonstrate a striking imbalance between pro- and antiinflammatory cytokines and a rise in the blood levels of electrolytes K+ and Na+ as well. The analysis performed revealed a direct correlation between the level of blood concentration of IL-6 and that of K+, Na+. PMID- 11881368 TI - [The medical and economic basis for optimization of the treatment and diagnosis of pneumonia patients under market conditions]. AB - Consideration is given to the optimization of the treatment-and-diagnostic process in patients with pneumonia under conditions of market relations, with particular regard to the development and perfection of the outpatient care, adoption of family medicine, and development of the orthospital care as well. PMID- 11881369 TI - [Assessment of efficacy of thrombolytic therapy in the treatment and prognosis of myocardial infarction]. AB - An apparent benefit has been shown to occur in acute myocardial infarction from the thrombolytic and antiaggregation therapy, that was evidenced by the absence of early post-infarction angina pectoris, decline in the incidence of disturbances of the heart's rhythm and conductivity, shortening of the hospital stay. PMID- 11881370 TI - [Experience gained with the use of local administration of medicinal remedies in rheumatic diseases]. AB - In the article, errors that are frequently encountered in dealing with rheumatic diseases are analysed together with the experience gained by the authors themselves with the management of 380 patients using local injections of corticosteroids (diprospan) and chondroprotectors (alflutop). The employment of local injection therapy has been shown to shorten considerably the patient's hospital stay, to improve the quality of life of the patients, with them being practically free from ill effects and complications. PMID- 11881371 TI - [Ultrasound densitometry of the heel bone in the diagnosis of osseous tissue metabolic abnormalities in patients with rheumatoid arthritis]. AB - The implication was studied of such factors as occurrence of the affliction, sex, inflammation activity, joint functional inadequacies, intake of steroids in the development of osteopenia and osteoporosis in patients with rheumatoid arthritis (RA). Determined in the above patients were RRF, SOS, BUA with the aid of the ultrasonic densitometer UBIS 5000 of the DMS (FRANCIUM) firm. Of the 42 examined patients, osteoporosis was identified in 28.5 percent, osteopenia in 45.3 percent of cases, the degree associated with RA duration, inflammation activity, function disorders of the joints favored by intensification of processes of osteal resorption in rheumatoid arthritis. PMID- 11881372 TI - [Lipid peroxidation abnormalities in miners with Reiter's disease and gout]. AB - The paper presents results of investigations designed to study changes in lipid peroxidation and in antioxidant system of miners in different vocational groups with Reiter's disease and gout. It is shown that the condition of the above systems is affected by both diseases as such and occupational conditions of miners. Based on the secured findings it is recommended that treatment of the above patients be supplemented with an antioxidant agent, copherol in particular. PMID- 11881374 TI - [Etiological diagnosis and classification of Herpes virus-associated disturbances of the central nervous system]. AB - An etiological classification is submitted of Herpes virus-induced affections of the central nervous system (CNS), such as monoherpesviral, herpesvirus-bacterial, herpesvirus-spirochetal, herpesvirus-mycotic, herpesvirus-protozoan affections. Excerpts from case records of etiologically confirmed mixed infections are given. Consideration is given to such herpesviruses as HSV, CMV, EBV, VZV. Other possible combinations of etiological agents include herpesvirus + chlamidia, mycoplasms, prions. PMID- 11881373 TI - [Clinical classification of iron deficiency anemia]. AB - Symptoms have been analyzed of clinical pleomorphism of iron deficiency anemia (IDA). In the examination of 89 patients with IDA, as example, a complex of secondary abnormalities of metabolism accompanying IDA clinical course is demonstrated. Based on the study of aspects of the clinical course, features of the peripheral blood, indices for iron metabolism, measuring of the blood content of lactic, pyruvic acids, free histamine, free serotonin, free heparin in the blood plasma a clinical classification has been elaborated detailing stages of the condition, basic clinical forms and complications. A classification of IDA is presented. Included in the above classification are stages of the above medical condition, its clinical forms and complications with diagnostic criteria and policy of dealing with IDA outlined. The use of such a classification ensures continuity of work among physicians when they come to deal with problems of diagnosis, treatment and prophylaxis of IDA in patients. PMID- 11881375 TI - [Innovative technologies and innovative policy in the health system of Ukraine]. AB - In the paper, consideration is given to the conceptual framework of the problem, as to which issue opinions vary among specialists. This has a negative impact on organization of carrying on of innovatory policy in the health service. Factographic data are presented on innovatory technologies in the health service, qualitative and quantitative parameters of such activities. The role is highlighted that the Ukrmedpatentinform has in the formation of the system of means of scientific communication, ways are considered for improvement of such a work, in the first place, at the expense of creation of feedback between "donors" and "recipients" of innovatory processes. It is emphasized that outside innovatory technologies scientific produce loses its value. Indices for innovatory activities are to be regarded as those of immediate importance in assessment of scientific and practical work of an institution. PMID- 11881377 TI - [Combination hypotensive therapy in patients with essential hypertension]. AB - In men and women with moderately severe essential hypertension (EH), efficiency has been analyzed of different schemes of combined hypotensive drug therapy depending on the hemodynamic type of circulation with taking account of basic indices for the 24-hour profile of arterial pressure (AP). It is shown that the baseline indices for a long-term AP monitoring (1-3 days) determine the choice of individualized differentiated therapy consisting of a combination of two (atenolol and enalapril maleate) or three (atenolol, enalapril maleate, hydrochlorothiazide) hypotensive drug preparations. Individual schemes of the above drug therapy permitted not only the correction of identified disturbances in the 24-hour AP profile but also contributed to the improvement of hemodynamic maintenance of graded physical loads, favouring positive changes in the structural-and-functional status of the myocardium, which facts suggest indirectly an improvement of prognosis in EH patients. PMID- 11881376 TI - [Classification of pneumoconiosis]. AB - The article presents a modern classification of pneumoconiosis taking account of the international classification and experience gained by specialists in this country. PMID- 11881378 TI - [Central hemodynamic status in young patients with myocardial ischemia]. AB - Results are submitted of investigations designed to study central hemodynamics and intracardiac kinetics in young patients with ischemic heart disease (IHD). In young coronary patients the course of the disease has been shown to be more often accompanied by the development of cardiac aneurism than in the elder ones in spite of the presence in the latter of more significant changes in their echocardiograms. Concomitant with aggravation of clinical manifestations of IHD (higher functional class of stable angina) there is an increase in the size of the left ventricle and decrease in the ejection fraction. It has been shown that hemodynamic parameters are closely related to both veloergometry indices and levels of cholesterol of high density lipoproteins APO-A-I. PMID- 11881379 TI - [Humoral immunological aspects of the pathogenesis of rheumatism, nonrheumatic carditis and juvenile rheumatoid arthritis in children]. AB - The state has been studied of the humoral component of immunological reactivity according to the content of the serumal complement, complement-bounding antibodies, immunoglobulins and immunoglobulin-antibodies of the main classes in the time-related course of rheumatism, nonrheumatic carditis and juvenile rheumatoid arthritis (JRA) in children. Tissue, enzymic, streptococcal antigens have been used together with those of nucleotides, nucleases. The obtained results suggest to us that the humoral component of bodily immunological reactivity has an important part in pathogenesis of the above medical conditions. It can be used in the diagnosis of affections of the heart. PMID- 11881380 TI - [Role of the skin in destabilization of coronary circulation]. AB - Our objective in this paper was to study diagnostic specificities of the skin secretion lipids fattyacid composition (FAC) in the precardiac region of patients with ischemic heart disease (IHD). Overall ninety two IHD patients who ranged from 34 to 78 years were examined together with fifteen clinically healthy persons. Used in the study were the gas-liquid chromatography techniques. The secured results suggested to us changes in the precardiac region skin lipids FAC in IHD. During the stage of compensation of coronary circulation the secretion of palmitic and docozohexaenic acids was found to have gotten diminished while that of saturated acids was augmented. This suggests to us a positive role of the skin, its negative role being in an increased secretion of monounsaturated fatty acids (MUFA), decreased secretion of representatives of omega-6 polyunsaturated fatty acids (PUFA). During the stage of decompensation the skin has predominantly a negative part (increased secretion of MUFA, docozohexaenic acids, decreased secretion of omega-6PUFA). PMID- 11881381 TI - [Responses of the glutathione system in patients with ulcerative pyloroduodenal stenosis before and after surgery]. AB - Studied among indices for the glutathione system in 99 patients was content of total, oxidized, reduced glutathione. The studies made suggested that in patients beyond 60 years of age there are profound inadequecies in the glutathione system manifested by an apparent reduction in total, reduced glutathione and augmentation of the oxidized form in the blood. Surgical treatment in the above patient population has not been found to be associated with bringing all studied forms of glutathione back to normal, which fact attests to the need for quest of drugs-antioxidants to be employed in the complex of postsurgery curative measures. PMID- 11881382 TI - [Serum surfactant activity in patients with chronic cryptogenic hepatitis]. AB - The paper represents results of use of dynamic interphase tensiometry of the blood serum in patients with chronic cryptogenic hepatitis. Studied in a comparative aspect in patients with varying degree of severity of the course of chronic cryptogenic hepatitis were features of indices for the blood serum dynamic surface tension and relations between them. The above technique is an important investigational tool to evaluate the course of chronic cryptogenic hepatitis. PMID- 11881383 TI - [Apoptosis of peripheral blood lymphocytes in patients with chronic viral hepatitis]. AB - The examined 68 patients with chronic viral hepatitis (CVH) demonstrated a high Fas/APO-1 (CD95) expression on the peripheral blood lymphocytes. It correlated with the serum level of the tumor necrosis factor alpha (TNF alpha). The above findings suggest to us the implication of apoptosis in CVH pathogenesis. Following administration of interferonotherapy treatments the number of lymphocytes in the presence of apoptosis has gotten decreased. PMID- 11881384 TI - [Endogenous intoxication, immunological reactivity and erythrocyte membrane permeability in patients with chronic myelogenous leukemia]. AB - Patients with chronic myeloid leukemia (CML) have been shown to present with apparent metabolic abnormalities, such as enhanced endogenous intoxication against the background of deterioration of the functional condition of the liver and decline in immune reactivity of the organism; the erythrocytic membrane permeability changes considerably. The data secured are discussed from the standpoint of using them as prognostic criteria of the course of CML. PMID- 11881385 TI - [Evaluation of hemodynamic changes in inflammatory hyperplastic processes in the thyroid gland with the aid of ultrasound color mapping]. AB - Ultrasound dopplerography techniques constitute an investigation of highly informative value, designed to study vascularization of pathological states of the thyroid gland. It permits the most effective and comprehensive evaluation of the organ circulation along the periphery of the capsule, in the parenchyma, in all pathological formations. This warrants the use of the method in diagnosing, in identification of the process dissemination beyond the organ into the surrounding tissues or regional lymph nodes. Colour Doppler-mapping of the thyroid gland is a highly sensitive method of investigation of neovascularization of the organ which comes to be changed in cancer depending on the stage of the process. PMID- 11881386 TI - [Aspects of thyroxin-bound globulin metabolism in preterm infants in certain pathological conditions]. AB - A comprehensive evaluation has been undertaken of the functional status of the thyroid gland in premature infants presenting with sepsis. The content of the thyroid hormones T4, T3 was studied together with aspects of the thyroxin-bound globulin metabolism that proved to be of diagnostic value in the evaluation of abnormalities of metabolism in those premature infants presenting with sepsis. PMID- 11881388 TI - [Effort tolerance in patients with angina and in those free from typical angina in isolated and multiple disturbances of the coronary arteries]. AB - Examined in the study were 169 patients with ischemic heart disease divided into two groups: group I comprised 105 patients with angina, group II was 64 patients who did not have typical stenocardia. All patients were subjected to exercise test and coronoangiography (CAG). Angina patients with transitory ischemia in a VEM-test have not been found to differ from patients without typical angina with transitory ischemia of the myocardium provoked by exercise. Transitory ischemia of the myocardium occurring in those persons free from typical angina has the same prognostic value as in patients with angina. PMID- 11881389 TI - [Dynamics of the indices of the prostacyclin-thromboxane system in adolescents with hypotensive type vegetovascular dysfunction exposed to hyperbaric oxygenation]. AB - As many as 185 juveniles aged between 11 to 14 years presenting with hypotensive type vegetovascular dysfunction have been examined to study the time-related changes in the blood content of prostacyclin and tromboxane under conditions of hyperbaric oxygenation. The control group copmrised 69 essentially healthy juvenile subjects the same age as those with dysfunction. Hyperbaric oxygenation has had a normalizing effect on indices for the prostacyclin-tromboxane system in those juveniles presenting with hypotensive type vegetovascular dysfunction, which fact can be explained by a drop in tenseness of mechanisms of the vegetative imbalance compensation. PMID- 11881390 TI - [Complex rehabilitation of patients operated on for complicated peptic ulcer]. AB - Results are submitted of sanatorium-and-health resort treatment of 104 patients operated on with the aid of the organ-sparing and resection techniques for complicated forms of gastroduodenal ulcers. An algorithm of therapeutic measures taking account of sanatorium-and-health resort treatment has been worked out. It allows to lessen considerably the postoperative complications, to shorten duration of temporary disability, to reduce permanent disability after resection of the stomach 3-fold. PMID- 11881387 TI - [Structural and functional features of the ovaries as a risk factor for ovarian cancer in infertile women]. AB - In examining 502 female patients with cancer of the ovaries (OC) infertility was identified in 21.9 percent of cases. The prevalent form of infertility was tubal infertility (92.7%), there were but a few instances of the endocrine form (1.8%). The infertile women were studied for their hormonal and receptorial status (receptors of estradiol and progesterone). It has been established that women with the endocrine variant of infertility present with more prominent hormonal inadequacies in the hypophysis-ovaries system and manifest changes in the receptorial status of the ovaries than those with tubal infertility; the findings in the latter have not been shown to be different from controls. In endocrine infertility OC is in fact not encountered, which fact can be related to morphological features of the ovaries giving origin to endocrine disturbances, and to a decline in the receptorial status and absence of ovulation. PMID- 11881391 TI - [Novel method for the surgical treatment of retropancreatic pseudocysts]. AB - There has been developed and adopted the author's method for surgical treatment of the retropancreatic cyst of the pancreas. The method involves a one-stage internal drainage of the cyst and pancreatic duct through cystoductopancreatojejunostomy on the isolated loop of the small intestine. Overall three patients have been operated on. The immediate and late results are good. The method is defended by the inventor's certificate. PMID- 11881392 TI - [Blood insulin levels in surgical treatment of peptic ulcer]. AB - Blood plasma insulin has been studied in 74 patients with gastroduodenal peptic ulcer (GDU) with the aid of the radioimmunological method before the operation, in the early and remote postoperative periods. Prior to surgery, the level of insulin gets increased on an empty stomach and after stimulation both in gastric and in duodenal peptic ulcer. The hormone content has been found to be reduced postoperatively, both in the immediate and remote periods but pyloroplasty in selective proximal vagotomy and especially in selective vagotomy resulted in a greater increase in insulin following its stimulation with test breakfast. Rehabilitative measures before and after surgery have been shown to be associated with augmentation of the blood level of the hormone, which fact promotes increase of the trophic effect of insulin on the gastrointestinal system. PMID- 11881393 TI - [Use of food additives in a multimodal treatment of disseminated pulmonary tuberculosis]. AB - Effects have been studied of multimodality treatment on the function of external breathing in patients with disseminated pulmonary tuberculosis using food additives Trace Minerals. Positive dynamics has been recordable of changes in indices for forced lung vital capacity, minute oxygen consumption, both prior and after loading, and maximum volume speed (MVS75) before loading. The obtained data are indicative of the efficacy of using vitamin-mineral additives Trace Minerals evidenced by improvement in bronchial patency and normalization of ventilation and-perfusion relations in patients with destructive forms of pulmonary tuberculosis. PMID- 11881395 TI - [Breast cancer in women with prior malignant tumors]. AB - Indices have been studied for a relative risk of development of subsequent metachronic breast cancer (BC) in 15627 sick female subjects presenting with different localizations of primary malignant tumours. The risk for development of BC in those women presenting with cancer of chief localizations exceeds the populational morbidity probability up to 30-fold. It is the female patients having cancer of the other mammary gland, endometrium, thyroid gland, lung, and skin who demonstrated the highest risk for development of BC. Such a polyneoplasia was recorded in 150 female subjects out of the 191 ones (78.6%). The risk for BC development in those female patients with cancer of the ovaries, cervical cancer, stomach, and melanosis cutis appeared to be insignificant. Metachronic BC in cancer patients depending on the site of the primary tumour can be a sequel of different pathogenetic mechanisms, of which hormonal-and-metabolic disturbances are the chief ones together with the genetical factor, and exposure to prior methods of antitumour chemoradiation therapy as well. PMID- 11881394 TI - [Prognosis of pretreatment outcome in patients with acute glomerulonephritis after an immunological review]. AB - Expediency has been shown for the immunological investigation to be done in those patients with acute glomerulonephritis in the nephrosis stage before the start of treatment to find out relevant approaches to such patients. The most informative parameters characterizing immunity include the peripheral blood absolute content T-lymphocytes, relative and absolute content of B-lymphocytes in the peripheral blood, IgA, IgG in blood plasma. However, to prognosticate near term clinical efficiency of treatment with immunodepressive drugs it is essential that all basic indices for the cell-bound and humoral immunity be taken account of. PMID- 11881396 TI - [Accelerated development of spinal degenerative changes during low back pain in young people]. AB - A possibility has been explored of an accelerated development of degenerative changes in the vertebral column in those young people whose activity is connected with high physical loads on the lumbar spine. A comparative analysis was done of results of the ultrasound investigation and magnetic-resonance tomography of spinal soft tissues. Sonographic techniques permit the conclusion to be reached that there develop degenerative and inflammatory processes at the level of certain vertebral segments at the preX-ray stage when it is only the soft-tissue structures that are involved. PMID- 11881397 TI - Cost survey offers bonanza of capitation benchmarks. PMID- 11881398 TI - HMOs still favor capitation, long-term trends show. PMID- 11881399 TI - Careful oversight helps CA group to profit under cap. PMID- 11881400 TI - HMO enrollment stabilizes, but number of HMOs continues to shrink. PMID- 11881401 TI - Do your patient satisfaction rates stack up to these national standards? PMID- 11881402 TI - [Chronic renal insufficiency. I: Definition, clinical course stages, progression mechanisms, etiology, and diagnostic criteria]. PMID- 11881403 TI - [Chronic renal insufficiency. II: Anamnesis and physical examination]. PMID- 11881404 TI - [Chronic renal insufficiency. III: Evaluation of the patient. Renal function tests. Complementary studies. Laboratory. Ultrasonography. Other imaging techniques. Renal biopsy]. PMID- 11881405 TI - [Chronic renal insufficiency. IV: Diagnostic algorithm. Diversion methods]. PMID- 11881406 TI - [Nephrologic clinical guidelines in primary care]. PMID- 11881407 TI - [Chronic renal insufficiency. V: Follow up of the patient]. PMID- 11881408 TI - [Chronic renal insufficiency. VI: Progression prevention]. PMID- 11881409 TI - [Chronic renal insufficiency. VII: Therapeutic algorithm. Possible problems and their solution]. PMID- 11881410 TI - [Chronic renal insufficiency. VIII. Comorbidity in renal insufficiency (RI)]. PMID- 11881411 TI - [Diabetic nephropathy: algorithms for evaluation and follow up. Diversion criteria. Therapeutic protocol]. PMID- 11881412 TI - [Assessment of integrated replacement therapy in patients with terminal renal insufficiency: selection vs election]. PMID- 11881413 TI - [Hypertensive nephropathy]. PMID- 11881414 TI - [The dialyzed patient: types, indications, follow up protocol, potential problems in primary care and their management]. PMID- 11881415 TI - [The transplanted patient]. PMID- 11881416 TI - [Use of drugs in renal insufficiency. Contraindicated drugs (absolute and relative contraindication)]. PMID- 11881417 TI - A decade of continuous improvement in cadaveric organ donation: the Spanish model. PMID- 11881418 TI - How can we improve organ donation rates? Research into the identification of factors which may influence the variation. PMID- 11881420 TI - [Lupus nephritis: treatments for today and tomorrow]. PMID- 11881419 TI - [The macrolaboratory and analytical quality in nephrology. A personal experience]. PMID- 11881421 TI - [Toward an evidence-based immunosuppression]. PMID- 11881422 TI - [Presentation of cancers in recipients of a solid-organ transplant]. PMID- 11881423 TI - [Structural changes and vascular calcifications in uremia]. PMID- 11881424 TI - [Nephropathy caused by polyomavirus type BK in renal transplantation]. PMID- 11881425 TI - [Renal effects of the chronic inhibition of nitric oxide synthesis in cirrhotic rats with ascites]. AB - Previous studies have shown that acute inhibition of nitric oxide (NO) synthesis improves sodium and water excretion and increases blood pressure in cirrhotic rats with ascites, thus suggesting that NO is an important factor contributing to the arterial hypotension and sodium retention of liver cirrhosis. In the present work we have analyzed the renal effects derived from the chronic oral treatment (10 days) with aminoguanidine (AG, 100 mg/kg/day), a preferential inhibitor of inducible NO synthase (iNOS), or Nw-Nitro-L-Arginine Methyl Ester (L-NAME, 0.5 mg/kg/day), a nonselective inhibitor of NOS, in an experimental model of liver cirrhosis with ascites (carbon tetrachloride inhalation). Untreated cirrhotic rats showed lower mean arterial pressure (MAP), diuresis, natriuresis and glomerular filtration rate (GFR) and similar renal blood flow (RBF) compared with the untreated control rats. Chronic administration of AG did not modify significantly any parameter in cirrhotic and control animals. Conversely, long term L-NAME administration to cirrhotic rats normalized MAP and significantly increased water and sodium excretion, whereas in control animals these parameters were not significantly modified. These results show that chronic NO synthesis inhibition with L-NAME, but not with aminoguanidine, improves renal perfusion pressure and increases the lower sodium and water excretion of cirrhotic rats with ascites. Thus, an enhanced production of NO is an important factor contributing to the renal sodium and water retention characteristic of liver cirrhosis. PMID- 11881426 TI - [Severe maternal complications associated with pre-eclampsia: an almost forgotten pathology?]. AB - Pre-eclampsia is a pregnancy-specific hypertensive syndrome associated with significant morbidity and mortality in mother and baby. With the increasing understanding of the disease process, the number of complications, and the maternal and perinatal deaths have fallen over the last few decades in the developed countries. In other parts of the world, the rates of mortality and morbidity remain high. We present eight cases of pre-eclamptic women with severe complications (eclampsia, HELLP syndrome, acute pulmonary edema, acute renal failure...) that were treated at our hospital in only a year, when we had not seen these pathologies in the last nine years. There was fetal death in three of the cases, related to abruptio placentae. There were no maternal deaths, but four mothers needed to be transferred to the intensive care unit, and required life support techniques. The causes of these new events remain unclear. Can they be due to increase in maternal age or to the high incidence of abruptio placentae? Or maybe can they occur only by chance? These women with pre-eclampsia and severe complications that can have potentially devastating consequences are not an easily identified group. We concluded that women with pregnancy hypertension must be carefully managed by expert physicians, particularly if they are more than 30 35 years old, overweight, with previous history of hypertension or nulliparous, in order to decrease these several complications. PMID- 11881429 TI - [Availability of permanent vascular access at the beginning of hemodialysis: role of the pre-dialysis consultation]. PMID- 11881428 TI - [Distribution of hepatitis C virus genotypes among the hemodialysis population in the province of Alicante]. AB - Hepatitis C virus (HCV) genotypes are irregularly distributed among the different geographic area and groups at risk. OBJECTIVE: To study the different HCV genotypes and subtypes of hemodialyzed patients from Alicante. METHODS: We studied 640 patients on haemodialysis (HD) and we determined the RNA-HCV and the genotypes in the 120 patients with antibodies against HCV (HCV-Ab). We compared the results with the genotypes of 1,370 patients from other groups at risk in the same geographic area. RESULTS: RNA-HCV was not found in the serum in 15% (18/120) of the patients on HD who were HCV-Ab positive. Prevalence of the different genotypes in the 102 patients with positive viral RNA was the following: 1b: 56.8% (58/102), 1a: 19.6% (20/102), 3: 17% (17/102), 2a-2c: 1.9 (2/102), 2b: 0.9% (1/102) 4: 2.9 (3/102), 5: 0.9% (1/102). In conclusion, the genotype 1b was the most frequent in the patients studied in all these areas, and was the same as in the rest of the country. This genotype has been associated with the most severe hepatic disease and poor response to treatment, affecting the prognosis of these patients. The most frequent genotypes in HD in Alicante were 1b, 3 and 1a. HCV genotypes distribution among the HD units was not uniform in the different geographic areas. HCV genotypes distribution in the HD population is similar to other groups at risk from the same geographic area. PMID- 11881430 TI - [Acute kidney failure caused by paracetamol poisoning]. AB - Paracetamol poisoning is manifested by hepatotoxicity, but acute renal failure is very rare, especially when there is no fulminant hepatic damage with encephalopathy or severe haemodynamic alterations. We present here the case of a 22-year-old woman who presented with acute renal failure after the ingestion of 11.5 g of acetaminophen. The clinical course and laboratory data were consistent with tubular necrosis. The patient required hemodialysis, but finally renal function returned to normal. The acetaminophen pharmacology and the differential diagnosis of acute azotemia in paracetamol overdosage are reviewed. PMID- 11881427 TI - [Clinical experience with icodextrin. Multicenter study]. AB - The aim of this study was to analyse our experience with icodextrin in Andalusia, Spain. The study includes 51 patients (30 women and 21 men) on peritoneal dialysis (21 on CAPD and 30 on Automated Peritoneal Dialysis) treated with icodextrin for 10.3 +/- 7 months (0-41 months). Their mean age was 57 +/- 18 years (18-86 years). We have recorded the appearance of side effects, and the evolution of several biochemical parameters at baseline and after 6, 12 ans 18 months from initiation of icodextrin. We also studied drainage fluid from 12 patients after an icodextrin exchange. RESULTS: There were side effects (all cutaneous) in 4 out of 51 patients (7.8%). Two of the affected suffered from cutaneous hypersensitivity reactions, and icodextrin had to be suspended; the other two had exfoliative dermatitis affecting hands and feet that disappeared without have to withdraws icodextrin. Biochemical parameters: Serum sodium levels decreased from baseline to six months (138 +/- 6 mEq/l vs 136 +/- 3 mEq/l; p = 0.006), and then persisted at the same levels throughout the rest of the study period. There was a slight but significant decreased of serum HDL-cholesterol at six months vs baseline (55 +/- 26 mg/dl vs 51 +/- 20 mg/dl, p = 0.04), and a further decrease at twelve months vs six months (42 +/- 15 mg/dl vs 51 +/- 13 mg/dl, p = 0.054). There were no significant variations of glucose, osmolality, cholesterol, LDL-cholesterol (tendency to increase), triglycerides, beta 2 m and weight (tendency to increase; p = 0.08). In relation with the icodextrin exchange: average ultrafiltration 296 +/- 119 ml (ranging from 104 to 480 ml), creatinine clearance 1.9 +/- 0.5 litres (20.5% of daily creatinine clearance), urea clearance 2.08 +/- 0.5 litres (18.7% of daily urea clearance), total protein losses 3.2 +/- 0.9 g, albumin losses 1.4 +/- 0.5 g; urea and creatinine clearances were negatively correlated with ratios D/P4 of urea and creatinine of PET and positively correlated with ratio G4/G0. In conclusion, side effects are scarce with the use of icodextrin. As described in other studies, there is a trend to a slight decrease in serum sodium. The long-term use of icodextrin does not-prevent weight gain or deterioration of patients on peritoneal dialysis, despite the diminution of glucose load. PMID- 11881431 TI - [Calciphylaxis: an uncertain pathogenesis and controversial treatment]. AB - Calciphylaxis is an uncommon but serious disease process that affects mainly patients with advanced renal failure. Calciphylaxis is characterized by dermal arteriolar calcification that leads to skin ulceration, necrosis, ischemia and secondary infection. The pathogenesis is poorly understood, although the calcium phosphorus product has been proposed as a major cause. Given the high morbidity and mortality rate, emphasis should be placed on prevention and early diagnosis of vascular calcification, as well as in prophylaxis of secondary infection. We present changing concepts in four patients receiving dialysis. PMID- 11881432 TI - [Acute kidney failure as the clinical presenting form of renal Burkitt's lymphoma in an HIV-positive patient]. AB - Burkitt's lymphoma is a tumour often associated with low immunity as acute lymphoblastic leukaemia (l3) or infection by the human immunodeficiency virus (HIV). The incidence of renal affection is variable (34-62%) and there are different aetiologies. We present a case of acute renal failure in a patient with a Burkitt's lymphoma and renal infiltration, and infected by the human immunodeficiency virus. PMID- 11881433 TI - [Is the index of response to erythropoietin (IRE) a good marker of adequate dialysis?]. PMID- 11881435 TI - [Primary biliary cirrhosis and minimal change glomerulonephritis]. PMID- 11881434 TI - [Fluconazole prophylaxis for fungal peritonitis in peritoneal dialysis]. PMID- 11881438 TI - Rethinking breast cancer. PMID- 11881436 TI - [Soft-tissue abscess caused by Rhodococcus equi in a patient on hemodialysis]. PMID- 11881437 TI - [Massive rhabdomyolysis associated with the use of cerivastatin monotherapy]. PMID- 11881439 TI - Smile--you're on botox! PMID- 11881440 TI - This bud's not for you. PMID- 11881441 TI - The chemistry of love. PMID- 11881442 TI - Faith in our fathers? PMID- 11881443 TI - Nightmare in Pleasantville. PMID- 11881444 TI - In the germ labs. PMID- 11881445 TI - How do you feel about nuclear power now? PMID- 11881446 TI - The $200 billion miscarriage of justice. PMID- 11881447 TI - Increasing the knowledge and understanding of philanthropy: the Center on Philanthropy at Indiana University. AB - The Center on Philanthropy at Indiana University's mission is to help non-profit leaders, fund raisers and others meet those needs by increasing knowledge and understanding of philanthropy--what it is, what motivates it, what impact it has- and teaching ways to improve its practice. PMID- 11881448 TI - Recognizing and building your organization's major gift potential. AB - Major donors should be exposed to all aspects of the organization and allowed to pick and chose their level and type of involvement. This means that the CEO and representatives should visit the donor to begin developing a personal relationship, including discovering their personal needs and interests. PMID- 11881449 TI - How to make an impression in a competitive market. PMID- 11881450 TI - Is it feasible? AB - The benefits offered by a feasibility study are considerable. Not only does it enable an organization to set a realistic campaign goal, but it can also help with the more inexact science of assessing the fund-raising climate. PMID- 11881451 TI - The non-profit sector and the market: opportunities & challenges. AB - By carefully examining the most important challenges and opportunities facing America's private non-profit organizations, the Non-profit Sector Strategy Group is working to stimulate a new consensus about the non-profit sector's roles and responsibilities, and offer practical recommendations to enhance policy, practice, research, and public education on this crucial set of institutions. PMID- 11881452 TI - [Effects of hyperbaric oxygenation and ceruloplasmin on redox processes and phosphorylation associated with them in the salivary glands in chronic nitrate intoxication]. PMID- 11881453 TI - [Two models for validation of caries etiology from a total systems approach viewpoint]. PMID- 11881455 TI - [Role of phagocytosis activation in the mechanism of therapeutic effect of medical ozone in patients with sluggish purulent inflammations of maxillofacial soft tissues]. AB - Sixty-three patients with local sluggish suppurative inflammations of maxillofacial soft tissues were examined. Medical ozone was added to the treatment protocols of 30 of these patients. Before treatment peripheral blood leukocyte phagocytic activity was decreased in 54 patients, which corresponded to clinical form of a sluggish inflammation. Traditional therapy did not appreciably change the level of phagocytosis. Addition of medical ozone exposure to therapy promoted normalization of leukocyte phagocytic activity and accelerated liquidation of inflammation. PMID- 11881454 TI - [Clinical and microbiological efficiency of silard gel-immobilized ethonium in the treatment of periodontal inflammations]. AB - In the course of examination and treatment of 250 patients studied the effect of medical sorbent, sylard-gel, and aethony immobilized on it upon inflammatory periodontal diseases. The results of the microbiological examination shaved good sorption and inhibitory characteristics of immobilized aethony. The best clinical results were achieved in the treatment chronic generalized gingivitis and chronic generalized periodontitis. PMID- 11881456 TI - [Frequency of removal of different teeth in persons aged 35-44 years]. AB - Observation covered 107 persons (41 males and 66 females) from 35 to 44 years of age. The results indicate that percentage of subjects with at least one-removed tooth is high even at the age 35-44 years (91.59%). Average number of teeth present (25.06), not counting third molars, gives reasons to confirm that the level of dental health is satisfactory. There is no considerable variation in the number of retained teeth between males and females. More often are missing first molars, followed by second morals and second biocuspids. Lower front teeth are the most often retained teeth. PMID- 11881457 TI - [Analysis of antibacterial activity of a new antiseptic for the treatment of maxillofacial inflammations]. AB - Clinical and experimental studies of polydimethyldiallylammonium chloride (PDMDAAC) were carried out in order to select the optimal concentration and composition of the preparation. 1% PDMDAAC solution showed high antiseptic activity and can be used for local therapy of the periodontal inflammations and infected posttraumatic wounds. PMID- 11881458 TI - [Efficiency of immune correction in patients with chronic traumatic mandibular osteomyelitis]. AB - The effect of a new immunomodulator neovir was studied in 35 patients with chronic traumatic mandibular osteomyelitis. The immune status and clinical parameters notably improved. PMID- 11881459 TI - [Laser exposure and noncoherent infrared therapy in the treatment of mandibular fractures]. AB - The efficiency of laser exposure and noncoherent infrared (IR) therapy in patients with mandibular fractures were compared. Non coherent IR exposure promoted a decrease in the number of inflammatory reactions and accelerated rehabilitation of patients. PMID- 11881460 TI - [Bone tissue morphological structure in congenital deformations of the jaws]. AB - Morphological structure of bone tissue was studied in various types of congenital deformations of the jaws. Morphological changes in the bone with deformations and the severity of these disorders depended not so much on the type on the deformation, but mainly on its severity, which can be explained by a drastic increase of functional exercise because of impaired occlusion and impossibility of proper chewing. Decelerated weak restructuring of bone tissue and imperfect osteogenesis in deformed bone, similar in various types of deformations, were demonstrated on morphological material. These changes can affect the regenerative potential of the bone in operated zones. PMID- 11881461 TI - [A comparative analysis of healing mandibular defects due to exposure to different plastic materials (experimental morphological study)]. AB - Autogenous bone grafts are the most effective factor able to activate osteogenesis in bone defects, as shown by our experiments. Experimental biomaterials such as Bio-Oss and KP-3 have a high osteogenesis potential, too. However the callus initiated in jaw defects by those biomaterials underwent partial reduction during secondary rebuilding, thus becoming a less authentic bone formation. PMID- 11881462 TI - [Comparative assessment of 5-year survival with cancer of the buccal mucosa treated by different methods]. AB - We analyzed case histories of 495 patients with oral cancer treated in the Head and Neck Tumor Clinic of the Russian Cancer Research Center RAMS from 1985 to 1998. 88 patients were included in the study out of which 50 reserved combined (radiotherapy + surgery) and 38--complex (chemotherapy + radiotherapy + surgery) treatment. On assessing the clinical results we got the following results: after combined treatment 5-year survival was 45.9 +/- 7.0%, while after the complex method--37.2 +/- 7.8%. The inclusion of chemotherapy to the treatment does not significantly influence long-term results. PMID- 11881463 TI - [Increase of resistance of dental abutment tissues to acid demineralization after preparation for fitting with permanent dentures]. AB - The efficiency of various protective treatments of teeth were studied in 50 patients divided into 4 groups: 1) (15 patients) 30 teeth with viable pulp were coated with fluorine varnish; 2) (15 patients) 30 teeth were examined after wearing temporary polypropylene crowns fixed on aqueous dentin; 3) (10 subjects) 30 teeth were examined after removal of permanent dentures and treatment of the enamel with fluorine-containing varnish; and 4) (10 subjects) after removal of permanent dentures the enamel was treated with calcium-containing gel using gingivodental kappa according to the proposed method. The status of dental hard tissues was evaluated using acid biopsy of the enamel (Leontyev and Distel's method in our modification) and analysis of the oral fluid. The efficiency of different methods of dental hard tissue protection from acid medium was evaluated. Positive results were obtained with pathogenetically justified new remineralizing procedure. PMID- 11881464 TI - [Simulation of stressed status of dental abutment tissues in normal and abnormal position of teeth]. AB - Based on the final elements method, the authors simulated the stressed-deformed state of dental abutment tissues in normal and abnormal position of the teeth. The distribution of normal and tangent tensions in dental transverse sections and their contact interactions with the alveolar process (periodontium-periosteum) were studied. The effect of the inclination angle of abnormally positioned teeth in various force of occlusion was demonstrated and maximum values of this force were established from the strength viewpoint. The results confirm the efficiency of the proposed scheme for estimation of the stressed-deformed status of dental abutment teeth in practical orthodontics. PMID- 11881465 TI - [New potentialities of laser therapy and electrostimulation after cheilorhinoplasty]. AB - A method for stimulation of the central regulation mechanisms has been used in 53 patients during the early period after cheilorhinoplasty. Electrostimulation by the flickering reflex method is a nonspecific method stimulating sanogenesis which can be used for repair of the neuromuscular function after correction of secondary deformation of the upper lip and palate. Transcerebral laser stimulation indirectly affected the reparative processes in the maxillofacial area by improving metabolism in the cerebrocortical projection zones. PMID- 11881466 TI - [Dental status of children whose parents are engaged in polyurethane foam manufacture]. AB - Children of workers engaged in polyurethane foam industry have high occurrence of cariosity and noncarious teeth affections, alteration of the mineral saliva composition, subnormal general and local immunity. PMID- 11881467 TI - [Superelastic properties of nickel-titanium arches made in Russia]. AB - The force-curvature relationship of nickel-titanium orthodontic arches manufactured by Kassis firm (Russia) were experimentally studied. The arches are characterized by pronounced superelastic properties. Theoretical prediction of forces developed by the Kassis Ni-Ti arches of any diameter with great curvature was made. The results are summed up in a table convenient for practical use. PMID- 11881468 TI - [Comparative characterization of the quality of life of patients with a dental profile]. PMID- 11881469 TI - [Why dentistry and its "section" maxillofacial surgery became a third-quality science?]. PMID- 11881470 TI - [Pathomorphology of regenerative processes in mandibular fracture after sodium succinate treatment and laser magnetotherapy in an experimental setting]. AB - Morphological reactions in tissue adjacent to mandibular angular fracture were studied in guinea pigs treated with sodium succinate and laser magnetotherapy. Due to succinate therapy the exudative component of inflammation was less expressed in comparison with the control, macrophagal reaction and neoangiogenesis were activated, the volume of damaged muscle tissue and the incidence of suppurations decreased. The number of osteoblasts increased and new bone structures acquired a lamellar pattern earlier than in the control. Sodium succinate therapy in combination with laser magnetotherapy had a more pronounced positive effect as regards activation of macrophagal reaction and neoangiogenesis and a decrease in the area of fibrosclerotic changes in the zone of damaged muscles, where newly formed myosymplasts differentiated into myotubes and even in muscle fibers. Suppuration of the wound was prevented. Bone tissue in the fracture zone formed without preliminary formation of cartilaginous tissue, which resulted in more rapid osteogenesis (lamellar bone growth in the fracture zone). PMID- 11881471 TI - [Ways of improving the system of registering and reporting system of the Russian Federation forensic medical laboratories]. PMID- 11881472 TI - [Evaluation of the provision and needs of the medical service of the Russian Federation for hospital beds]. AB - Taking into account the average statistical data of the last years the need of treated contingents for hospital beds constitutes 9.7%. Increase in accessibility of hospital treatment for the military pensioners and members of families as well as improvement of management of military hospital affairs will increase the index up to 9.9%. With transfer of military medical service to national standards and development of hospital-substitutive technologies the need for hospital beds will constitute 8.4%. The results obtained show that if all contingents attended to RF AF medical service would have the opportunity to treat only in the military medical institutions the reforming of military health should be aimed at maintenance and more effective use of hospital bed fund. PMID- 11881473 TI - [Plastic surgical correction of defects of long tubular bone defects with free blood supplied autografts]. AB - Treatment of the patients with long tubular bone defects is the difficult problem. At present the most widespread bone graft methods allow to achieve the result but not always satisfy physicians and patients. The use of revascularized bone autografts opens the new opportunities in the treatment of such patient category. Case records of 34 patients with long tubular bone large defects who underwent the bone graft with revascularized bone autograft were analyzed. Good and satisfactory anatomical and functional results were achieved (follow-up was up to 10 years), some complications and possible methods of their prevention were analyzed. PMID- 11881474 TI - [Features of arterial hypertension in participants of regional military conflict]. PMID- 11881476 TI - [Organizational principles of military health care in the Russian Federation]. PMID- 11881475 TI - [Choice of proton pump inhibitors]. AB - The criteria of choice of proton pump inhibitor at acid-dependent diseases such as gastroesophageal reflux, peptic ulcer, diseases associated with Helicobacter pylory are discussed in the article. The presented data of meta-analysis of efficiency (clinical-and-endoscopic, economical), safety and tolerance, cost and simplicity of use show that today rabeprasol (pariet) is the drug of choice. PMID- 11881477 TI - [Clinical picture, early diagnosis and treatment of abdominal typhus in servicemen in the Russian Army in Tadzhikistan]. PMID- 11881478 TI - [On reactogenicity and immunogenicity of "Euvax B" in children with chronic diseases]. PMID- 11881479 TI - [Experience in studying antigenic activity of inactivated hepatitis A vaccine]. AB - The results obtained during the study of antigeneity of inactivated vaccines against hepatitis A ("Avaxim", France; "Hep-A-in-VAC", Russia; "Havrix", Belgium) used in troop units and RF MD institutions are presented in the article. The aim of the investigation is choice of the drug that provides the production of antibody protective levels in maximally short periods. It is very important for the army collectives sent to the zones of armed conflicts and HAV epidemic centres. It is shown that the most acceptable vaccine is "Avaxim" that provides the highest percentage of seroconversion (93.75%) with antibody level not less than 125 mIU/ml. It is recommended to consider this level as the protective one in conditions of army collectives. PMID- 11881480 TI - [An outstanding clinician and military-field surgeon (on the 100th anniversary of the birthday of I.S. Kolesnikov)]. PMID- 11881481 TI - [Outstanding domestic scientist-epidemiologist (on the 80th anniversary of the birth of V. D. Beliakov)]. PMID- 11881482 TI - [In the central military specialized hospital]. PMID- 11881483 TI - [The role of endotoxin of gram negative bacteria in the pathogenesis of atherosclerosis]. AB - Literature data and the results of own research on the role played by lipopolysaccharide (endotoxin) of Gram negative bacteria in the initiation and progressing of atherosclerosis are summarized. Endotoxin promotes activation of all cells taking part in the formation of atherosclerotic plaques, induces the transformation of macrophages of arterial intima into foam cells as well as endothelial lesions and hyperlipidemia, reduces the ratio of cholesterol in lipoproteins of high specific density to total blood cholesterol. With chlamydiae and enterobacteria used as examples, the role of Gram negative bacteria and their endotoxin in the etiology and pathogenesis of atherosclerosis is discussed. PMID- 11881485 TI - [Problems of the taxonomy and nomenclature of bacteria of the genus Salmonella]. AB - The main stages of the development of the nomenclature and classification of salmonellae are analyzed. Modern problems and different points of view of microbiologists on the methods of their solution are presented in detail. The acceptability of some projects is discussed. PMID- 11881484 TI - [Characterization of Vibrio cholerae eltor isolates according to their epidemic potential using new diagnostic cholera bacteriophages eltor ctx+ and ctx- and by the polymerase chain reaction]. AB - The epidemic potential of 113 V. cholerae eltor strains of different origin was determined with new diagnostic cholera bacteriophages eltor ctx+ and ctx-, as well as the test for hemolytic activity. Of these strains 50 were epidemically safe and 51 were epidemically dangerous, while the epidemic potential of 12 other strains could not be detected. Determination of genes ctxA, tcpA and toxR in the strains under study by means of the polymerase chain reaction (PCR) revealed that epidemically dangerous strains carried the whole set of the above genes in 92.2% of cases. 98.0% of epidemically safe cultures were lacking either gene ctxA, or genes ctxA and tcpA, or genes ctxA, tcpA and toxR, which confirmed their incapacity to cause cholera. The results of the differentiation of the cultures with new diagnostic cholera phages coincided with the results of PCR in 90% of cases. The most complete and reliable evaluation of the epidemic potential of individual vibrio isolates may be obtained using the two compared methods. The amplification test system gives more information when isolates with unclear epidemic potential are analyzed. PMID- 11881486 TI - [Center of special laboratory diagnostics and treatment of dangerous and exotic infectious diseases in the system of the antiepidemic protection of the territory of the Russian Federation]. PMID- 11881487 TI - [Interaction of the vegetative and nonculturable forms of Salmonella typhimurium with bacteria of the genus Bdellovibrio]. AB - The data on the interaction of bacteria of the genus Bdellovibrio with the representatives of pathogenic Salmonella typhimurium are presented. Different types of such interaction are demonstrated: in a two-component system, in fluid media, in an agar layer and on the surface of a solid carrier. As shown for the first time, Bdellovibrio cells are capable of interacting not only with actively growing bacteria, but also with their noncultivable forms. The data obtained may serve as the basis for the study of possible practical use of such bacteria for controlling Gram-negative organisms, the causative agents of sapronotic infections. PMID- 11881488 TI - [Stimulating and inhibiting effect of Spirulina platensis on microorganisms]. AB - The blue-green microalga (cyanobacterium) S. platensis and the complex of its metabolites in the culture fluid being added into nutrient agar in doses of 0.01, 0.1 and 10 mg/ml, may produce a stimulating and inhibiting effect on microorganisms. In Spirulina biomass and in culture fluid substances which may be associated with the stimulation or inhibition of the growth of some microorganisms are supposedly contained. The manifestation of stimulating or inhibiting properties depended on the concentration of S. platensis and the complex of its metabolites, as well as on sterilization methods of nutrient media supplemented with these substances. PMID- 11881489 TI - [Etiological structure of acute respiratory viral infections in children hospitalized in 1981-1999]. AB - The role of influenza and parainfluenza viruses, respiratory syncytial viruses and adenoviruses in the etiological structure of morbidity in acute respiratory virus infections (ARVI) in children hospitalized during the 19 year period is analyzed. As the result of examination of 56,287 patients by direct immunofluorescent test, respiratory viruses were detected in 21% of cases. The seasonal character, periodicity and level of ARVI morbidity were established. According to medical records, in the 1990s ARVI took a more severe course in children than that observed in the 1980s. In addition, the data on morbidity among children regarding rotavirus infection and ARVI were found to be similar. PMID- 11881490 TI - [Analysis of polymorphism of the interleukin-4 gene of healthy and HIV-infected persons]. AB - The distribution of the allel variants of the promoter area (C = 590T) of the interleukin-4 (IL-4) gene in HIV-infected and relatively healthy representatives of the Caucasoid population has been studied. The relationship between the genotypes of this polymorphism and the production of IL-4 by mononuclear cells of peripheral blood as well as distribution of IL-4 genotypes among males and females is analyzed. The occurrence of the homozygous combination of the allel variant C/C of the promoter of IL-4 has been shown to prevail almost twofold over the occurrence of the variant C/T among healthy donors and HIV-infected patients. Sexual differences play an essential role in the character of inheriting the allel variants of the genes of IL-4, the presence of the homozygous variant C/C or T/T being a risk factor of HIV infection in males. As revealed in this study, in the peripheral blood of healthy donors mononuclear cells having genotype C/C differ from cells with the heterozygous variant C/T in higher spontaneous production of IL-4 and, simultaneously, in lower capacity for the activation of its production in response to stimulation with mitogen. In HIV-infected patients mononuclear cells differ in higher spontaneous production of IL-4 in comparison with controls. We may thus infer that the human genotype controlling the initial level of the production of IL-4 by lymphocytes Th2 may influence the intensity of antibody production in the process of infection. PMID- 11881491 TI - [Studies on halophilic vibrios causing a food poisoning outbreak in the city of Vladivostok]. AB - V. parahaemolyticus and V. alginolyticus strains isolated from patients during an outbreak of an acute enteric disease in Vladivostok in 1997 were studied. All strains were found to possess typical taxonomic signs. V. parahaemolyticus isolated from humans had direct heat stable haemolysin exotoxin. The overwhelming majority of these strains belonged to serovar O3K6. Among the cultures under study 7 phage types were determined: phage types 1, 2, 7, 10 in 8 V. parahaemolyticus strains and phage types 2, 4. 5. 7 in 5 V. alginolyticus strains. The diagnostic halophilic phage lyzed vibrios in 30.2% of strains. The cultures under study were found to be highly sensitive to chloramphenicol, cefotaxime, nalidixic acid and cyprofloxacin. The study proved that the outbreak of alimentary toxicoinfection was caused by vibrios of serogroup O3:K6. PMID- 11881492 TI - [Ecological-genetic mechanisms of the transition of Salmonella typhimurium to the dormant state in the environment]. AB - The dynamics of vegetative and dormant (noncultivable) of S. typhimurium cells of two isogenic strains in association with microalgae Scenedesmus quadricauda and under the action of the exometabolites of the algae at different stages of their growth was studied using in parallel bacteriological method and PCR. The study revealed that at the stage of active growth green algae and their metabolic products maintain the survival of salmonellae (strain TR = 1) vegetative forms in water at an optimum temperature. Low temperatures induced their gradual (3 weeks) transition to the dormant state. The exometabolites of old dying algae induced the rapid (several hours) and complete transition of the bacterial population (TR = 1) to noncultivable state. In our experiments the insertional mutation in gene pqi (strain PhoA = 8), inducing the defect of transmembrane protein and disturbances in the transition of salmonellae to dormant state, led to stable existence (lasting 7 months, i.e. the whole term of observation) of vegetative cells. The natural inducers tried in our experiments did not lead to the formation of the dormant forms of salmonellae in this mutant strain. PMID- 11881493 TI - [Specific prophylaxis of influenza in organized groups of children with the vaccine Vaxigrip]. AB - The results of the open comparative study on epidemiological effectiveness and tolerance of the vaccine Vaxigrip, carried out in 109 children aged 2-15 years with different initial state of health, are presented. The index of epidemiological effectiveness of Vaxigrip was 4.09 (p < 0.05), the rate of epidemiological effectiveness--75.6% (p < 0.05). Vaxigrip was found to be well tolerated by both healthy children and those with different functional disturbances and chronic diseases. The study revealed that the use of Vaxigrip was not accompanied by any systemic postvaccinal reactions. High prophylactic effectiveness and good tolerance of Vaxigrip make it possible to recommend it for mass immunoprophylaxis of influenza in children. PMID- 11881495 TI - [Specific features of diphtheria vaccinal process in children with rheumatic diseases]. AB - Clinical and immunological examination of 55 children aged 6-15 years with rheumatic diseases, immunized against diphtheria, was carried out. All children were immunized at the stage of clinical and laboratory remission and in some cases while undergoing a prolonged course of cytostatic therapy or therapy with nonsteroid anti-inflammatory remedies. This examination demonstrated that in the overwhelming majority of children with rheumatic diseases the diphtheria vaccinal process took an asymptomatic course and had no influence on the course of the main disease. Specific features, characteristic of the immune status of this group of children, were established. In the course of the vaccinal process the restoration of the initially inhibited characteristics (the production of TNF alpha and IL-2) to normal values were shown to occur, which was indicative of the fact that the reserve capacities of immunocompetent cells were retained in these patients. This study also revealed that immunization of children with rheumatic diseases with adsorbed DT and D toxoids with reduced antigen content was not excessive antigenic stimulation for such children, as it did not lead to immunopathological shifts, but induced transient phase changes in immunological characteristics, similar to those in healthy children. Protective levels of antibodies to diphtheria were shown to retain for a long time with considerable prolongation of intervals between booster injections. The simultaneous course of immunosuppressive maintenance therapy in the average dosage used for the corresponding age group did not inhibit the production of protective antibodies. PMID- 11881494 TI - [Humoral response to the injection of acellular Pertussis vaccine]. AB - The humoral response of mice and rabbits to the injection of whole-cell pertussis vaccine (PV) and acellular pertussis vaccine (APV), developed at the Mechnikov Research Institute for Vaccines and Sera (Russian Acad. Med. Sci.) in Moscow, was studied. In the sera of immunized animals antibodies to the antigenic complex were determined in the direct hemagglutination (DHA) test, specific antibodies to filamentous hemagglutinin (FHA) and pertussis toxin (PT)--in the enzyme immunoassay (EIA) and antibodies neutralizing PT in a cytopathogenic dose (CPD)- in neutralization test on Chinese hamster ovary (CHO) cells. In mice and rabbits immunized with APV the antibody titers determined in the DHA test were higher than those in the animals immunized with PV. Specific antibodies titers to FHA and PT in the sera of rabbits immunized with APV were also higher than those in the sera of rabbits immunized with PV. High dilutions of sera taken from the animals immunized with APC neutralized 4-16 doses of PT in the neutralization test on CHO cells. The most important result of this study was the detection of a more pronounced immune response in the animals immunized with APV in comparison with that induced by PV according to the results obtained in EIA and in the test of PT CPD neutralization on CHO cells. PMID- 11881496 TI - [Reactions to and immunogenic properties of the combined vaccine Bubo-M for immunization of children against diphtheria, tetanus and viral hepatitis B]. AB - Bubo-M, the first Russian combined vaccine, was found to have low reactogenicity. The difference between the number of postvaccinal reactions in the group of children immunized with Bubo-M (25.9%) and those in the group of children who had been simultaneously injected into different sites of the body with ADS-M toxoid (adsorbed DT toxoid with reduced antigen content) and hepatitis B vaccine (26.7%) was not statistically significant. Following immunization a considerable increase in the level of diphtheria and tetanus antibodies (p < 0.005) was observed in all children (100%), the level of HBs antibodies in the group of children immunized with Bubo-M (the geometric mean titer: 13,721 IU/l) essentially exceeding that observed in the control group injected with ADS-M toxoid and hepatitis B vaccine (the geometric mean of the titer: 2,441 IU/l). Bubo-M was duly registered and allowed for industrial production and medical use. PMID- 11881497 TI - [Autoantibodies in children with chronic inflammatory lung diseases]. AB - 124 sera of children with chronic bronchitis, chronic pneumonia, bronchial asthma, exogenic allergic alveolitis, congenital developmental defects of the lungs and the syndrome of the situs inversus of organs were examined with a view to study the state of humoral immunity to tissues. The study was carried out by means of the enzyme-linked immunosorbent assay with the use of collagen, elastin, DNA (native and denaturated), membrane antigens of the lung, the liver, the small intestine and the large intestine. Among all groups of patients autoimmune disturbances, manifested by a rise in the level of autoantibodies of different specificity, were registered. The degree of manifestation of autoimmune disturbances depended on the kind of pathology. After treatment a decrease in the level of autoantibodies was registered in the examinees. PMID- 11881498 TI - [Functional activity of endotoxin binding factors in chronic viral hepatitis B and C]. AB - The functional state of cell-mediated and humoral antiendotoxin factors in patients with chronic viral hepatitides B and C has been studied. A decreased content of antibodies to glycolipid of chemotype Re and to Escherichia coli O14 with common enterobacterial antigen was shown to occur in these diseases. In addition, a decreased number of neutrophils, eosinophils and thrombocytes has been noted. The conclusion has been made that patients with chronic hepatitides are not protected from the pathogenic action of endotoxin penetrating from the intestines into the systemic blood stream. PMID- 11881499 TI - [Investigation on the mechanisms of leukocyte alteration in patients with intolerance to some drugs (with novocaine as a model)]. AB - The mechanisms of novocain damaging action on blood cells in persons with increased sensitivity to this drug were studied in the leukocyte alteration test. The leading role of histamine liberated from basophils was established. The increased sensitivity to novocain was shown not to be passively transferred to the cells of healthy donors with patient serum with intolerance to the drug; moreover, the joint action of both cell-mediated and thermolabile humoral factors was found to be necessary for the realization of leukocyte alteration under the action of novocain. The comparison of the information content of a number of methods--skin testing, dosed provocation, leukocyte alteration test and chemiluminescence--for revealing increased sensitivity to novocain in 30 persons with adverse reactions to this preparation registered in their medical history. The two in vitro tests were shown to be comparable in their diagnostic significance with the method of dosed provocation. PMID- 11881500 TI - [Approaches to prevention and treatment of latex allergy]. AB - Data on sensitization to latex as well as measures aimed at prevention and treatment of latex allergy, are presented. The intensity of the symptoms manifestation of latex allergy was shown to depend on the duration of contact with latex. To prevent the development of latex allergy, the following preparations were used: the antihistaminic preparation Claritine, the immunocorrecting preparations Ruzam and polycomponent vaccine VP-4. The use of Claritine was shown to lead to the alleviation of the symptoms of latex allergy, but after treatment with Claritine was stopped the symptoms of latex allergy reappeared. The clinical effect lasted for as long as 2 months after treatment with Ruzam, while in case of polycomponent vaccine VP-4 use remission was registered even 3 months later. The data presented thus confirm topicality of the latex allergy problem and practical importance of using the immunocorrecting preparations Ruzam and polycomponent vaccine VP-4 for its prevention and treatment. PMID- 11881501 TI - [Intestinal microbiocenosis at the critical periods of child development]. AB - The bacteriological study of fecal specimens obtained from 595 children at different critical periods of the development of their immune system was carried out. The comparison of intestinal microflora in children at the critical periods of the development of their immune system revealed that in children under 1 year considerable changes in the microflora composition were observed during the first 6 months of life. These changes, most pronounced in children aged 1 month and 3-6 months, were characterized by decreased level of indigenous microflora and an increase in the qualitative and quantitative content of aerobic microflora with the representatives of the family Enterobacteriaceae playing the dominating role. In boys such disturbances were manifested to a greater extent than in girls. PMID- 11881502 TI - [Isolation and identification of Crimean-Congo hemorrhagic fever virus in the Stavropol territory]. AB - The results of the isolation and identification of the causative agent of a haemorrhagic fever outbreak in the Stavropol Territory are presented. The virus isolated from blood of haemorrhagic fever patients by virological methods was identified in serological and molecular tests as Crimean haemorrhagic fever virus. This epidemiological analysis testify to increased activity of the natural focus of Crimean-Congo haemorrhagic fever in this area due to a number of natural and other factors leading to intensification of its epidemic realization. PMID- 11881503 TI - [Dysbioses of the oral cavity and intestines and immune reactivity in of adolescent bronchial asthma patients]. AB - Sixty bronchial asthma (BA) patients aged 15-18 years were examined between attacks. The composition of the oral cavity and intestine microflora as well as the total IgE level, peripheral blood eosinophils and lymphocytes expressing receptors CD3, CD4, CD8, CD22, CD11b, CD71, HLA-DR were detected. Colonization of the oral cavity by Staphylococcus aureus in 9%, by hemolytic streptococci in 10%, by fungi of the genus Candida in 25% and by enterococci in 7% of the patients was revealed. From the intestines S. aureus were isolated in 21%, hemolytic Escherichia coli in 18% and fungi of the genus Candida from 45% of the patients, while lactic acid bacteria were isolated only in 39% of the patients. The patients of this group were found to have eosinophilia (4.21 +/- 0.05%, p < 0.001) and a high concentration of IgE (330.2 +/- 29.31%, p < 0.01); in these patients the index CD4/CD8 was lower (< 0.05), the number of CD11b-, CD71- and HLA-DR positive lymphocytes was elevated. Thus, at periods between attacks the examined BA patients were found to have dysbiosis of mucous membranes, an elevated content of IgE, eosinophilia and lymphocyte activation indicative of the absence of immunological remission and required adequate diagnostic and therapeutic measures. PMID- 11881504 TI - [Adjuvant properties of subalin, a recombinant interferon-producing probiotic]. AB - The adjuvant properties of subalin, a recombinant probiotic prepared from live bacteria Bacillus subtilis producing human alpha 2-interferon were studied in the scheme of its use with vaccines against parvovirus enteritis and distemper. Subalin was shown to be capable of preventing immunosuppression caused by the injection of vaccines, accelerating the formation of the antigen-specific clone of memory cells and enhancing antigen-specific immune response. The mechanisms of the adjuvant effect of subalin were considered; this effect was shown to be due to the action of interferon excreted by bacteria of B. subtilis into the lumen of the intestine. The advantages of this method of interferon supply and the prospects of using subalin preparation as adjuvant are discussed. PMID- 11881505 TI - [Polymorphism of human genes and HIV infection]. AB - Literature data on the possible interrelation between resistance to human immunodeficiency virus (HIV) infection as well as rapidity of progressing of clinical manifestations of HIV infection and the presence of certain mutations in the human genome is discussed. Special attention is paid to the polymorphism of genes coding receptors of chemokines. PMID- 11881506 TI - [Role of dysbacteriosis in the formation of chronic noninfectious pathology in children]. AB - The results of clinical observations and laboratory data make it possible to regard dysbacteriosis as an important factor in the pathogenesis of chronic noninfectious pathology in children. The adequate complex correction of intestinal dysbacteriosis on the basis of probiotic therapy facilitates the prolonged remission of the disease in children with diabetes mellitus of type 1 (DM1) and myopathy, decreases severity of late complications of DM1. A suggestion is made on the role of dysbiotic microflora in the development of chronic non infectious pathology in children. PMID- 11881507 TI - [House-dust mites and mold fungi as a source of indoor allergens]. AB - In this work information on the biology and ecology of synanthropic mites and mold fungi, proved to be the sources of indoor allergens in different premises, is presented. The authors propose the algorithm of the ecological monitoring of dwellings, developed on the basis their investigations. The types of dwellings, the time and situations when ecological monitoring should be carried out are analyzed. Grounds for the necessity of controlling the number of mites and the concentration of Micromyces spores, as well as their elimination from the environment, are given. PMID- 11881508 TI - [Culture media as artificial environment ensuring the growth and development of microorganisms]. AB - Literature review is presented on culture media that are regarded as artifical environment for microorganisms permitting modeling of various conditions for revealing cultural, morphological and biochemical properties, characteristic for a given microorganism, as well as manipulations with these conditions. PMID- 11881509 TI - Seeking balance in complexity. PMID- 11881510 TI - Shaping the advanced practice psychiatric-mental health nursing role: a futuristic model. AB - As advanced practice psychiatric-mental health nursing has transitioned from earlier models of practice, elements of clinical specialist and psychiatric nurse practitioner roles are being blended to produce a new type of practitioner. The challenge of preserving mental health expertise while expanding advanced practice primary and primary mental health care competencies is addressed in several nursing education models. At New York University's Division of Nursing, faculty have designed a program around elements identified as essential to the autonomy demanded of the evolving role, knowledge, and skills basic to broad based health care and mental health care delivery with quality patient care outcomes and the competencies necessary for accountability as care providers in a changing health care delivery system. Essential elements, resources to identify them, and strategies to attain them are discussed. Approaches that promote student, clinician, and faculty development and maximize education affirm the specialty's capacity for innovation and the profession's capacity for new direction and futuristic change. PMID- 11881511 TI - Distance education: pedagogy for psychiatric-mental health nursing. AB - Education of undergraduates in mental health nursing necessitates modeling interpersonal relationships and providing accessible educational experiences. This article describes (1) a distance education strategy that promotes interpersonal learning, and (2) a qualitative study exploring faculty's experience with this pedagogy. Several themes emerged from the data: I Feel Like I'm In A Different World; I Don't Get To Know The Students; They Have My Face; I Am Caught Up In The Technology; It Is A Lot Like Teaching In The Classroom; and It Is Helping Me Grow In My Teaching. A metaphor, "bringing it to life," captures faculty members' capability to change lifeless lecture content into a learning experience involving human beings. PMID- 11881512 TI - "She looked just like me." A domestic violence learning module. AB - This paper describes a 22-hour domestic violence learning module that is incorporated into the psychosocial course for seniors in a baccalaureate nursing program. As part of their learning experiences, students attend circuit court, meet with judges, and accompany advocacy workers. Additionally, they attend group therapy sessions with both the victims and perpetrators of abuse. Students keep journals reflecting their thoughts, feelings, and reactions throughout the experiences. Thematic analysis of these journal entries revealed five common themes. Students recognized their encounters in clinical situations as frightening and emotionally difficult, expressed surprise at their reactions to perpetrators, identified with victims, wrestled with issues of good and bad, and reported that stereotypes about victims and perpetrators had been incorrect. Debriefings and support by faculty are important for students throughout the experience. PMID- 11881513 TI - Case-based teaching and learning experiences. AB - Case-based teaching and learning strategies can be utilized to assist advanced practice psychiatric nursing students in both obtaining requisite knowledge and enhancing clinical reasoning skills. We discuss the benefits of case-based learning in terms of how it (1) contributes to students' appropriate organization of information to be recalled later for use in clinical reasoning situations; (2) generates experiences that students would not otherwise have; (3) increases the visibility of students' clinical reasoning processes; and (4) enhances students' confidence. This article also explores three examples of case-based teaching and learning: use of written cases in a seminar; use of standardized patients in an assessment course; and utilization of web-based cases for learning assessment and intervention skills. Finally, we compare and contrast each of these methods in terms of their relative effectiveness in achieving each of the benefits. PMID- 11881514 TI - Turning danger into opportunity: teaching psychiatric nursing in the aftermath of a disaster. AB - In the wake of a devastating flood that destroyed the ecology of a small town, undergraduates taking their first clinical course in psychiatric mental health nursing formed AGAST (Awesome Guerillas Against Situational Trauma) and, with their instructor, performed individual and group crisis intervention over an intensive 90-hour clinical rotation. Three theories were used to help guide the students assessments and interventions with survivors. The clinical experience required innovation, close connection with the host community, and multi-level support from the educational institution. Students rose to the occasion and blossomed in the rich clinical matrix of the experience. The example can be applied to the use of other naturally-occurring events as theaters for innovative clinical teaching. PMID- 11881515 TI - Psychiatric-mental health nursing at the interface: revisioning education for the specialty. AB - In the emerging health care environment, families and communities are facing a broad range of conditions that require more integrated views of wellness and illness, mind and body, and the biomedical and the behavioral. This also calls for an enhanced care delivery system. Three interfacing areas of practice have particular significance for the psychiatric-mental health nursing workforce: enhanced primary care, public health or population-focused health interventions, and managed care. This paper addresses the broad range of knowledge, skills, and competencies that are required for the future of advanced practice psychiatric mental health nursing and describes interactive learning experiences and new pedagogies to deliver them. PMID- 11881516 TI - First incest disclosure. AB - Despite the pivotal importance of disclosure to incest treatment and healing, disclosure has never been studied from the victim's perspective. How do incest victims move from keeping the secret to speaking about their abuse? Nine adult women were asked to talk about the first time they each told about the incest. They often spoke, not of "telling" in the commonly understood sense (i.e. giving information to someone who understands one's meaning), but instead of a time when some form of knowledge of the incest first entered an interaction with another person. Colaizzi's (1978) phenomenological method was used to analyze the interviews. Seven themes emerged: (1) living in the silencing home; (2) I am totally and particularly alone; (3) my mother, the focus of need; (4) incest as burden; (5) the secret must be kept; (6) disclosure: trying to balance above a chasm; and (7) disclosure as loss: no matter what, I still lose. The themes were then integrated into an essential description of first incest disclosure. Implications for nursing practice are explored. PMID- 11881517 TI - A comparative study of the spiritual perspectives and interventions of mental health and parish nurses. AB - Spirituality is an integral part of holistic nursing practice. Limited research has been done that explores nurses' spirituality and the spiritual interventions they have made with patients in their practice. Much of the extant research has been done on nurses involved in terminal care such as oncology and hospice nursing. This study explores spiritual perspectives and spiritual nursing interventions in two other nursing specialties that require holistic nursing care as well: mental health and parish nurses. The findings indicate that both groups report high spiritual perspective scores and provide a variety of interventions to patients in their practices. PMID- 11881518 TI - Mental health services for persons with mental retardation: role of the advanced practice psychiatric nurse. AB - The term dual diagnosis is frequently used by mental health professionals to refer to people who have mental illness complicated by substance abuse. A less commonly recognized population are those individuals who have the dual diagnosis of mental retardation and mental illness. In this paper the author discusses (1) the mental health needs of persons with mental retardation and (2) the current state of services for them. There is a significant lack of appropriately trained professionals to help address the needs of this population. Traditionally, nurses have primarily been identified as providers of physical care. Advanced practice psychiatric nurses, however, are an underutilized group of professionals who have the capacity to assume a leadership role in clinical care, service coordination, and advocacy for individuals and families affected by mental illness and mental retardation. PMID- 11881519 TI - Evaluating improvements in nursing staff at a state psychiatric hospital. AB - In response to federal intervention in 1990, Hawaii initiated a number of changes at the state's psychiatric hospital. The process of change that was undertaken by the hospital's department of nursing from 1990 through 1994 to meet federal mandates was described in an earlier article in this journal. The present study evaluated the degree to which court-ordered improvements in nursing staff at the hospital were actually achieved between 1995 and 1997. Seven quantitative measures of compliance with federal mandates were evaluated, based on the court orders. Significant improvements were found on six of the seven measures during the 30-month study period. These included increases in nursing staff as measured by (1) the number of staff and (2) full-time equivalents, as well as increases in (3) the hours of nursing care per patient, (4) the number of staff per patient on each shift, and (5) the percent of RNs per shift. Use of agency personnel decreased significantly, as ordered, but overtime use increased significantly, contrary to court orders. PMID- 11881520 TI - Needs for assistance and emotional aspects of caregiving reported by AIDS patient caregivers in a day-care unit in Brazil. AB - The advent of AIDS in families, following the chronic evolution of the illness, has produced crisis situations unknown until now. With the objective of knowing this new reality better, 20 caregivers of AIDS patients were interviewed in southeastern Brazil. The collected data were analyzed using a qualitative method. The results showed difficulties regarding caregiving, such as a lack of orientation, materials, transport, and people to share the role of caregiver, as well as emotional difficulties, such as fears from infection, revulsion, pity, and impotence in the face of death. The role of families is essential in patient assistance, due to the inability of the health system to attend to all AIDS patients. Nevertheless, these families have to receive support mainly from nurses, in order to be helped in executing an aid plan to the ill person at home. Nurses and health care. PMID- 11881521 TI - Use of acamprosate and different kinds of psychosocial support in relapse prevention of alcoholism. Results from a non-blind, multicentre study. AB - OBJECTIVE: To determine abstinence rates and to obtain safety data during the use of integrated therapy with acamprosate and the main established psychotherapeutic intervention programmes in the treatment of alcohol dependence. DESIGN: Non blind, parallel-group, multicentre, phase IV study. SETTING: Naturalistic setting reflecting the clinical use of acamprosate in outpatient treatment of alcoholics in Germany. PATIENTS: 753 patients meeting the DSM-III-R criteria for alcohol dependence. At study entry, patients had to be abstinent for 1 to 4 weeks, and those who relapsed within the first 14 days of treatment were withdrawn from the study. INTERVENTIONS: All patients received acamprosate 1332 to 1998 mg/day according to bodyweight and were assigned to one of the following treatment groups: individual psychotherapy, group psychotherapy, behavioural therapy, brief intervention or family therapy. The patients were followed for 24 weeks. OUTCOME MEASURES: The principal outcome criterion was the time to the first drink. Secondary outcome measures were cumulative abstinence duration and craving. Adverse events were recorded systematically and classified according to World Health Organization adverse reaction terminology. RESULTS: The mean time to first drink was 81.5 days. 236 (33.5%) patients were continuously abstinent during the study. The discontinuation rate in the study was 49%, and the cumulative abstinence duration was 61%. Abstinence and discontinuation rates in this study were similar to those found in placebo-controlled clinical trials. Craving fell to scores of less than 50 on the Lubecker Craving Scale within 2 weeks of treatment initiation. There were no significant differences between the different psychosocial support treatment arms concerning time to first relapse or abstinence rates. Linear regression analysis showed severity of alcohol dependence according to DSM-III-R criteria to be associated with treatment outcome (p = 0.003). The most common adverse clinical events during the first 2 weeks of treatment were diarrhoea (69 patients), pruritus (37 patients), headache (28 patients) and fatigue (16 patients). Acamprosate was discontinued in nine patients (1%) because of severe adverse events; only two cases were clearly linked to treatment (diarrhoea and dermatitis). One death occurred during the study (status asthmaticus). CONCLUSIONS: Abstinence rates with integrated acamprosate and psychosocial support in a naturalistic setting are similar whatever the psychosocial support used, and similar to those seen in placebo controlled clinical trials of acamprosate. The safety profile of acamprosate was good. PMID- 11881522 TI - Acamprosate. PMID- 11881523 TI - Neramexane. PMID- 11881524 TI - Alefacept. Amevive, BG 9273, human LFA-3/Igg fusion protein, LFA 3, LFA 3 TIP LFA 3/CD2, LFA-3/Igg fusion protein, LFA3TIP, recombinant LFA-3/Igg1 human fusion protein, recombinantly engineered LFA-1/Igg1 human fusion protein. PMID- 11881525 TI - Aripiprazole. Abilitat, OPC 14597. PMID- 11881526 TI - Bevacizumab. Anti-VEGF monoclonal antibody, avastin, rhumab-VEGF. PMID- 11881527 TI - Dapsone--topical. Atrisone. PMID- 11881528 TI - Daptomycin. Cidecin, Dapcin, LY 146032. PMID- 11881529 TI - Efalizumab. Anti-CD11a monoclonal antibody--Genentech/Xoma, HU 1124, hu1124, xanelim. PMID- 11881530 TI - G 3139. Augmerosen, Bcl-2 antisense oligonucleotide--Genta, GC 3139, Genasense. PMID- 11881531 TI - IDM 1. IDM-1, MAK/IDM-1. PMID- 11881532 TI - Iseganan. IB 367, protegrin IB 367. PMID- 11881533 TI - Oxycodone/ibuprofen. PMID- 11881534 TI - Pitavastatin. Itavastatin, Nisvastatin, Nk 104, Nks 104, P 872441. PMID- 11881535 TI - Ramoplanin. A 16686, a 16686a, MDL 62198. PMID- 11881536 TI - Rimonabant. SR 141716, SR 141716a. PMID- 11881538 TI - Policing the "wild west" world of Internet pharmacies. PMID- 11881537 TI - Satraplatin. BMS 182751, BMY 45594, JM 216. PMID- 11881539 TI - Putting a new face on managed care. Consumers need convincing about quality issues. AB - Widespread consumer suspicion about the quality of managed care indicates that marketers still have work to do Research shows that women identify personal relationships with physicians, medical expertise, speed of access, and comprehensiveness of service as important signs of quality when selecting a managed care plan for their families. Based on existing research findings and new focus group data, this article provides guidelines for promoting an association of quality with managed care, particularly among women. PMID- 11881540 TI - Do ads really drive pharmaceutical sales? The true effects of DTC advertising remain a mystery. AB - The growth in mass media drug advertising has coincided with a rapid rise in spending on prescription drugs in the United States. Our research examines the potential link between increased ad spending and the rising use and sale of these drugs. We found strong circumstantial evidence that ad spending is one of many important elements driving the sharp rise in pharmaceutical sales to consumers. PMID- 11881541 TI - Generation Rx.com. What are young people really doing online? AB - The Kaiser Family Foundation examined the ways Generation Xers use the Web to find health information and found some surprising results. Not only do young adults access online health information, but they seek it more often than they check sports scores, purchase merchandise, or participate in a chat room. They're also likely to be influenced by what they find online, and many report that they've changed their behavior based on what they've learned. PMID- 11881542 TI - Ten steps to a successful trade show. A little planning can help prevent common marketing mistakes. PMID- 11881543 TI - Bringing cardiology services to a new market. A predictive index helps hospitals get to the heart of their customer base. PMID- 11881544 TI - Health care gets personal. An individualized approach can reap big rewards online. PMID- 11881545 TI - Beyond focus groups. A changing market demands nontraditional marketing approaches. PMID- 11881546 TI - Information to die for. Federal mortality data can be a gold mine for researchers. PMID- 11881547 TI - Pharmaceutical marketing in a new age. Effective campaigns still need to focus on what customers want. AB - The pharmaceutical industry has focused heavily on marketers' ability to market new products more efficiently. However, a more streamlined marketing approach can help address customers' needs and ease the pressure on drug companies to discover new drugs with blockbuster appeal. Through discussion and a detailed example, this article describes a stream-lined approach to creating more effective marketing and sales force strategies. PMID- 11881548 TI - [New diagnostic possibilities and new organization of therapeutic offers in multiple endocrine neoplasia]. PMID- 11881549 TI - [Diagnosis of infectious diseases. Nocardiosis]. PMID- 11881550 TI - [Microscopic colitis]. AB - Microscopic colitis is an umbrella term for a newly described group of colitides, belonging to the inflammatory bowel diseases, which are only diagnosable by microscopic evaluation of a macroscopically normal colon mucosa. Collagenous colitis and lymphocytic colitis are the most common of these colitides. Microscopic colitis is characterised clinically by chronic non-bloody watery diarrhoea. Crampy abdominal pain, nocturnal diarrhoea, urgency, and initial weight loss are usual. Concomitant diseases of autoimmune origin and arthralgia are commonly seen. Treatment of microscopic colitis follows the guidelines for treatment of other inflammatory bowel diseases, but a substantial part of the patients with microscopic colitis enter spontaneous remission after some years. A minor part, however, have very troublesome symptoms and are almost refractory to treatment. Microscopic colitis has apparently no malignant potential. PMID- 11881551 TI - [Pycnodysostosis--common ancestor of some Danish patients. Examination and diagnosis based on molecular genetics]. AB - Eight patients with pycnodysostosis from six Danish families were examined for mutations in the cathepsin K gene. Three different mutations are the cause of pycnodysostosis in the six families--five of whom come from Ringkobing County and one from Vejle County. One mutation has a high frequency in the families from Ringkoebing County. The five families are related through a common ancestor, who introduced the mutation around the year 1100. The disease is described with respect to aetiology, symptoms, prognosis, diagnosis, and symptomatic treatment. Research in pycnodysostosis may bring important knowledge to the understanding of related diseases, such as osteoporosis. PMID- 11881552 TI - [Smoking and asthma]. AB - Cigarette smoking is a well-known health hazard, probably not least for patients suffering from asthma. This review gives a short overview of the effects of passive and active smoking on the inception and outcome with of longitudinal changes in the lung function and mortality of patients with asthma. Substantial evidence suggests that smoking affects asthma adversely. Exposure to environmental tobacco smoke, especially maternal smoking in children, may be a significant risk factor for asthma. Such exposure in patients with established asthma is not only associated with more severe symptoms, but also with a poorer quality of life, reduced lung function, and increased utilisation of health care including hospital admissions. Active smoking does not appear to be a significant risk factor for asthma, but is associated with a worse outcome with regard to both longitudinal changes in lung function and asthma-related mortality. Based on current knowledge, it is therefore of utmost importance to encourage patients with asthma not to smoke, and accordingly these patients should be given full support in their right to a smoke-free environment. PMID- 11881553 TI - [The postgraduate training of general practitioners in communication and counseling. A questionnaire survey in the county of Aarhus]. AB - INTRODUCTION: The study aimed to describe the postgraduate training of the general practitioners (GPs) in communication and psychiatric counselling. MATERIAL AND METHODS: GPs in Aarhus County, Denmark, received a mailed questionnaire about psychiatric hospital training, participation in courses and Balint groups (psychiatric supervision), and their need for further training. RESULTS AND DISCUSSION: The questionnaire was returned by 320 (74.4%) GPs. Almost all GPs had received some kind of postgraduate training although to a very varying extent. Almost half had taken courses of more than three days' duration, and half were members of a psychiatric supervision group. Two-thirds of the GPs thought they needed further training. The need was independent of the GP's evaluation of his/her own psychiatric education. PMID- 11881554 TI - [Selection of patients referred to gastroscopy from general practice]. AB - INTRODUCTION: The aim of this study was to investigate whether guidelines for selecting dyspeptic patients for early endoscopy are appropriate and whether referrals from general practitioners give all necessary information. MATERIALS AND METHODS: We carried out a prospective study of consecutive referrals from general practitioners. The patients were referred by general practitioners to the Department of Medical Gastroenterology, Aalborg Hospital, during the period 1 February 1999 to 31 December 1999. The referrals for endoscopy were examined for information about the duration of symptoms, alarm symptoms (anaemia, dysphagia, vomiting, and weight loss), usage of ASA/NSAID, and medical treatment with acid suppressants. At endoscopy, similar information was recorded on a standardised form for comparison. RESULTS: Two hundred and ninety-nine patients, 150 men and 149 women, were entered in the study. The medium age was 51.3 years (17-95). Ninety-six (32%) patients had organic dyspepsia (ulcer, oesophagitis, cancer). Of 192 (46%) patients selected for early gastroscopy, 88 (46%) were assigned to early examination solely because of age > 45 years, and 21 (11%) solely because of alarm symptoms. All the patients with information about ASA/NSAID medication were older than 45 years. The diagnosis of cancer and ulcer was significantly more often found in the group of patients selected for early gastroscopy 31/192, as compared with the waiting list patients 9/107 (p = 0.004). DISCUSSION: The guidelines for distribution to early and waiting list endoscopy proved to be well suited for priority selection of patients with the highest risk of organic dyspepsia. PMID- 11881555 TI - [Attitudes and expectations of pregnant women to routine ultrasonography in early pregnancy]. AB - INTRODUCTION: More than 90% of pregnant women in Denmark are offered ultrasonography early in pregnancy to detect the viability of the fetus, multiple pregnancy, gestational age, and the locality of the placenta. Few hospitals offer ultrasonography to detect anomalies of the fetus, including chromosome abnormalities. The aim of this investigation was to elucidate the women's wishes and expectations concerning ultrasonography early in the pregnancy and the level of information on what the ultrasonography comprises. MATERIAL AND METHODS: In 1996, 1400 questionnaires were consecutively handed out to all pregnant women at their first attendance at the antenatal clinic run by the midwives in the County of Vejle. RESULTS AND DISCUSSION: One thousand and fifteen (72.5%) answered the questions anonymously. An overwhelming majority looked at sonography as a huge experience (86.9%), felt more secure about the pregnancy after sonography (80.4%), wished and expected to have a sonography for anomalies (84.3%) and were ready to have an elected abortion should a serious anomaly be detected (67.6%). In most hospitals in Denmark it is not possible to comply with the attitudes and expectations of the pregnant women concerning prenatal care in the form of ultrasonography. A thorough description of what the ultrasonography comprises is mandatory. PMID- 11881556 TI - [Growth in Greenland. Development of body proportions and menarcheal age]. AB - INTRODUCTION: No recent investigations of child growth in Greenland are available. Owing to the secular trend, earlier investigations are of limited clinical value. For this reason I have studied together with my staff the most important anthropometric measurements of schoolchildren in Maniitsoq. MATERIAL AND METHODS: We recorded the weight, standing height, sitting height, armspan, and menarcheal age. After the exclusion of 52 children with at least one parent not of Greenlandic origin and the exclusion of ten children who were either suffering from conditions known to influence growth or in whom some of the anthropometric data were lacking, 266 boys and 271 girls remained in the study. The 537 children were between 5 and 19 years old. RESULTS: Up to the age of 14, Greenlandic boys follow the Danish curves for standing height, then flatten out to a final mean height about 7 cms below the Danish level. The girls' growth curves follow the Danish ones only to the age of 11 years, then flatten out to a final height 5 cms below the Danish level. A somewhat higher sitting height ratio was recorded for both sexes compared to Danish children. The mean menarcheal age was 12.64 years. DISCUSSION: Growth curves show that up to the age of 14 Greenlandic boys now follow the Danish ones, thereafter flattening out. Girls' growth curves flatten out already at the age of 11. The final height for boys is 7 cms and for girls 5 cms below that in Denmark. During the course of 100 years menarcheal age in Greenland has been reduced by three years, and is now three months below that in Denmark. PMID- 11881557 TI - [Angiosarcoma of the adrenal gland]. AB - We describe a case of angiosarcoma of the adrenal gland in a 71-year-old man. Angiosarcomas are collectively one of the rarest forms of soft tissue neoplasms. They comprise less than 1% of all sarcomas. The primary treatment of choice is radical surgery. PMID- 11881558 TI - [Management of stomas]. PMID- 11881559 TI - [Mental health of elderly immigrants]. PMID- 11881560 TI - [Driver's licence and health-related suitability to drive registered motor vehicles]. PMID- 11881561 TI - [Antiepileptic agents and congenital abnormalities--induced abortions should be included]. PMID- 11881562 TI - [Is it allowed to write anything to promote screening?]. PMID- 11881563 TI - [Is new prescription of antibiotics after treatment with sulfonamide or pivmecillinam a validated method for assessment of treatment failure?]. PMID- 11881564 TI - [Error in an article on rectal cancer]. PMID- 11881565 TI - [Standards of examination requirements]. PMID- 11881566 TI - Fifty years of the "feeding center": not yet satiated. PMID- 11881567 TI - Review of Prof. B.K. Anand's scientific study: fifty years following his discovery of feeding centre. AB - Prof. BK Anand the living legend, is the founder of modern neurophysiology in India. His career spanned an era that marks the beginning of Brain Research in India. His contributions to understanding of brain functions began with the epoch making discovery of lateral hypothalamic area as the 'feeding center'. Subsequently it encompasses a wider range of studies from elucidation of the intricate mechanisms underlying ingestive behaviour to hypothalamic regulation of cardiovascular, respiratory, gastrointestinal activity and regulation of various physiological functions by limbic system. The following review gives an account of the contributions made by Prof Anand and his colleagues during twenty five years of his illustrious career. The publications resulted from his studies has not only stood as a testimony to understanding of the neural control of ingestive behaviour and limbic functions but also have been a sources of a great interest to those who pursue knowledge in this field. PMID- 11881568 TI - Membrane Na+ K+ ATPase inhibition related dyslipidemia and insulin resistance in neuropsychiatric disorders. AB - There are several reports in literature implicating cholesterol metabolism in the pathogenesis of neuronal degenerations, oncogenesis, functional neuropsychiatric disorders and multiple sclerosis. Biosynthesis of cholesterol takes place by the isoprenoid pathway, which also produces digoxin, an inhibitor of membrane Na(+) K+ ATPase. Inhibition of this enzyme results in intracellular Mg++ deficiency which can influence cholesterol metabolism. Digoxin also influences transport of tryptophan and tyrosine which are precursors of various neurotransmitters. Alterations in digoxin, membrane Na(+)-K+ ATPase and also in neurotransmitters have been reported in the disorders mentioned above. In view of this, serum lipid profile, activity of plasma HMG CoA reductase (the major rate limiting step in the isoprenoid pathway), RBC membrane Na(+)-K+ ATPase activity, serum Mg++ concentration, concentration of digoxin and concentration of serum neurotransmitters were studied in some neuropsychiatric disorders. The serum serotonin level was increased while that of serum dopamine and noradrenaline was reduced. Serum digoxin levels were high and RBC membrane sodium-potasium ATPase activity and serum magnesium were reduced. There was a reduction in HDL cholesterol and increase in plasma triglycerides (pattern similar to insulin resistance and syndrome X) in most of the disorders studied. The HMG CoA reductase activity was high, the serum total cholesterol was increased while RBC membrane cholesterol was reduced in most of the cases. The significance of increased digoxin with consequent inhibition of membrane Na(+)-K+ ATPase in relation to changes in cholesterol metabolism and insulin resistance type of dyslipidemia is discussed in this paper. PMID- 11881569 TI - Clitoria ternatea (Linn) root extract treatment during growth spurt period enhances learning and memory in rats. AB - Neonatal rat pups (7 days old) were intubated with either 50 mg/kg body weight or 100 mg/kg body weight of aqueous root extract of Clitoria ternatea (CTR) for 30 days. These rats were then subjected to open field, two compartment passive avoidance and spatial learning (T-Maze) tests (i) immediately after the treatment and (ii) 30 days after the treatment, along with age matched normal and saline control rats. Results showed no change in open field behaviour, but showed improved retention and spatial learning performance at both time points of behavioural tests, indicating the memory enhancing property of CTR which implicates a permanent change in the brain of CTR treated rats. PMID- 11881570 TI - Lipid peroxidation in haemorrhagic shock and after transfusion of blood in dogs. AB - The present study was carried out on mongrel dogs. Haemorrhagic shock of different severities and duration was produced by exsanguination from an artery. After the required duration of shock, two third of the volume of blood withdrawn was transfused back into the animal. Effect of haemorrhage and reperfusion of blood after haemorrhagic shock on lipid peroxidation was assessed by measuring plasma malondialdehyde (MDA). Severity of shock was assessed from the haematocrit values. There was a significant increase (P < 0.05) in plasma MDA level after blood transfusion in a group having 40 mm Hg blood pressure as magnitude of shock and one hour as duration of shock (Group II) only. Haemotocrit value was also significantly low (P < 0.05) in this group after haemorrhagic shock. Results are suggestive of lipid peroxidation with ischaemic reperfusion in severe and long duration of shock. PMID- 11881571 TI - Evaluation of E-721B, an indigenous herbal combination in experimental models of immediate hypersensitivity. AB - E-721B, an indigenous herbal combination was investigated for its usefulness in immediate hypersensitivity using different animal models. The drug inhibited the mast cell degranulation induced both by antigen and compound 48/80, the Schultz Dale response in sensitized guinea pig ileum smooth muscle preparation and the production of precipitating antibodies in 50% of tested rats. It also inhibited the mast cell degranulation in passively sensitized rats indicating its suppressive action on production of reaginic antibody (IgE). However, the drug did not inhibit the 48 hours passive cutaneous anaphylaxis reaction in rats, indicating that a single dose of the drug does not have cromoglycate like properties. All the above results indicate the inhibitory effect of E-721B on immediate hypersensitive reactions such as asthma. PMID- 11881572 TI - Neurobehavioural study of subchronic administration of oxydemeton-methyl (insecticide and acaricide) in rats. AB - Oxydemeton-methyl, an organophosphate insecticide and acaricide produced decrease in the exploratory behaviour and prolongation of barbitone sodium induced hypnosis in rats after intermittent aerosol spray inhalational exposure, for 1/2 hour daily for 7 consecutive days, compared to the saline control group. Further, ED50 +/- SEM value for haloperidol induced catalepsy, CD50 +/- SEM value for pentylenetetrazole induced seizure and CI50 +/- SEM value for electroshock (i.e. the dose of haloperidol, PTZ and intensity of electroshock producing catalepsy or positive seizure response in 50% of rats) were significantly decreased after 7 days exposure to oxydemeton-methyl compared to that of saline control group. The study has established the central nervous system depressant effect, extrapyramidal effect and proconvulsant potential of oxydemeton-methyl which is widely used by the agricultural workers in the form of field spray. PMID- 11881574 TI - Effect of Garcinia cambogia extract on lipids and lipoprotein composition in dexamethasone administered rats. AB - Dexamethasone (10 mg/kg body weight/day, s.c.) administered rats were treated with or without Garcinia cambogia fruit extract (1 g/kg body weight/day, orally) for 8 days. The administration of dexamethasone resulted in marked increase in the levels of triglycerides and cholesterol and free acids in both plasma and liver. The level of phospholipids increased in the plasma but decreased significantly in liver tissue after dexamethasone administration as compared to those in normal rats. The activities of lecithin cholesterol acyl transferase and hepatic lipoprotein lipase were lowered significantly after dexamethasone per se administration. The levels of HDL-triglycerides and HDL-cholesterol remained unchanged, while the LDL and VLDL increased significantly in dexamethasone administered rats. The lipid levels were maintained at near normalcy when co treated with Garcinia cambogia extract in dexamethasone administered rats. This study reveals the undesirable changes in lipid profile on dexamethasone administration and the hypolipidemic property of Garcinia cambogia extract. PMID- 11881573 TI - Effect of cancer treatment modalities on serum lipids and lipoproteins among women with carcinoma of the breast. AB - Serum lipids and lipoproteins were studied in 61 breast cancer patients before initiation of therapy and subsequently during and after completion of cancer therapy. Different serum lipid fractions were estimated by enzymatic method. It was observed that mean levels of serum triglycerides, total cholesterol and low density lipoprotein (LDL)-cholesterol among untreated breast cancer patients decreased significantly after treatment. On the contrary, an increasing trend in the levels of high density lipoprotein (HDL)-cholesterol was noticed in patients during the course of treatment. This study reflects the effects of cancer therapy in the alteration of levels of different serum lipid fractions in the patients with breast cancer. PMID- 11881575 TI - Improved performance in the Tower of London test following yoga. AB - Twenty girls between 10 and 13 years of age, studying at a residential school were randomly assigned to two groups. One group practiced yoga for one hour fifteen minutes per day, 7 days a week, while the other group was given physical training for the same time. Time for planning and for execution and the number of moves required to complete the Tower of London task were assessed for both groups at the beginning and end of a month. These three assessments were separately tested in increasingly complex tasks requiring 2-moves, 4-moves and 5-moves. The pre-post data were compared using the Wilcoxon paired signed ranks test. The yoga group showed a significant reduction in planning time for both 2-moves and 4 moves tasks (53.9 and 59.1 percent respectively), execution time in both 4-moves and 5-moves tasks (63.7 and 60.3 percent respectively), and in the number of moves in the 4-moves tasks (20.9 percent). The physical training group showed no change. Hence yoga training for a month reduced the planning and execution time in simple (2-moves) as well as complex tasks (4, 5-moves) and facilitated reaching the target with a smaller number of moves in a complex task (4-moves). PMID- 11881576 TI - Improvement in hand grip strength in normal volunteers and rheumatoid arthritis patients following yoga training. AB - The present study aimed at assessing the effects of a set of yoga practices on normal adults (n = 37), children (n = 86), and patients with rheumatoid arthritis (n = 20). An equal number of normal adults, children, and patients with rheumatoid arthritis who did not practice yoga were studied under each category, forming respective control groups. Yoga and control group subjects were assessed at baseline and after varying intervals, as follows, adults after 30 days, children after 10 days and patients after 15 days, based on the duration of the yoga program, which they attended, which was already fixed. Hand grip strength of both hands, measured with a grip dynamometer, increased in normal adults and children, and in rheumatoid arthritis patients, following yoga, but not in the corresponding control groups, showing no re-test effect. Adult female volunteers and patients showed a greater percentage improvement than corresponding adult males. This gender-based difference was not observed in children. Hence yoga practice improves hand grip strength in normal persons and in patients with rheumatoid arthritis, though the magnitude of improvement varies with factors such as gender and age. PMID- 11881577 TI - Auditory evoked potential responses in menopausal women: a normative study. AB - Three types of auditory evoked potential responses i.e. auditory brainstem response (ABR), middle latency response (MLR) and slow vertex response (SVR) were studied in 22 post-menopausal Indian women to have a normative value for each response. Recordings were done on computerised evoked potential recorder using 10/20 system of electrode placement and standard click stimuli. The results were comparable with similar studies done in the western world indicating that there is no ethnic variation in evoked potential responses. The latencies of ABR waves showed an increase in menopausal females when compared with young adults indicating a delay in neural transmission which might be due to changed hormonal milieu of sex hormones after menopause. The normative values of MLR and SVR which represent the thalamo-cortical and cortical auditory projections are being reported in Indian menopausal women for the first time. PMID- 11881578 TI - Effect of addition of proton carriers in culture medium on growth and secretion of hybridoma cell line OKT3. AB - Monoclonal antibodies (MAb) constitute the centre of all in-vitro diagnostic measures and almost all in-vivo therapeutic manoeuvres now. Production emphasis for these antibodies is having a current shift from animal-based large-scale culture to in-vitro bioreactor-based high-density culture. One of the major difficulties in high-density culture is end-metabolite accumulation in batch and fed-batch cultures in the forms of H+, NH4+ etc.. thereby reducing cellular growth and secretions. In the present study, effects of added proton carries--NAD and NADP--over and above the metabolic pools of the molecules, were examined on the cellular growth and secretion kinetics. Although NADP fortification showed a remarkable improvement in cellular growth (time dependent 200-300% improvements compared to controls) and size, cumulative MAb titre was better with NAD fortification. Combined additional loads of the proton carriers would be interesting to study in high density culture conditions. PMID- 11881579 TI - The role of hypo-osmotic swelling test in recurrent abortions. AB - The role of male factors in abortions has not been studied extensively. We undertook this study to determine if there was any relationship between hypo osmotic swelling test score and recurrent abortions. This was a cross-sectional case control study conducted at the semen analysis laboratory at a tertiary level referral centre. The male partners of 25 patients who had three or more first trimester abortions of unknown etiology were chosen as cases. Controls were 26 unmarried volunteers with unknown fertility potential to represent the entire population. The conventional seminal parameters were studied according to WHO guidelines. The HOS test score were obtained by the method advocated by Jayendran et al. There were no statistically significant differences in mean sperm count, percentage motility and morphology among the cases and controls. We found a low hypo-osmotic swelling test score among the cases [55.7 +/- 1.197 (SEM)] than the controls [69.3 +/- 1.143 (SEM)] which was statistically significant [P < 0.001]. The normal spermatozoal membrane is the prerequisite for the specialized cell-to cell communications and cell-to-cell binding. In spite of apparently normal semen analysis, subtle membrane defects in the spermatozoa, which could be the cause of defective membrane functions in the embryo effecting miscarriages, can be elucidated by the hypo-osmotic swelling test. PMID- 11881581 TI - Effect of alpha-MPT on imipramine induced antinociception in rats. PMID- 11881582 TI - Ophthalmoscopic changes in colour defective school children. PMID- 11881580 TI - Effect of Liv-52 an ayurvedic preparation on the pharmacokinetic profile of carbamazepine in monkeys. PMID- 11881583 TI - Flumazenil is a benzodiazepine receptor antagonist. PMID- 11881584 TI - Luteal phase in adolescent females. PMID- 11881585 TI - International collaboration in palliative care. PMID- 11881586 TI - Brugada syndrome--10 years later. PMID- 11881587 TI - [Acute confusion (delirium)in geriatric patients: a neglected interdisciplinary problem]. PMID- 11881588 TI - Infections in the elderly: Part 1. PMID- 11881589 TI - Bringing real life to the table. Patient reps help FDA review products. PMID- 11881590 TI - Why drugs get pulled off the market. PMID- 11881592 TI - First-of-a-kind heart device cleared. PMID- 11881591 TI - Losing weight: more than counting calories. PMID- 11881593 TI - Diabetes. A growing public health concern. PMID- 11881594 TI - Continuous-wear contact lenses approved. PMID- 11881595 TI - FDA approves first biologic treatment for sepsis. PMID- 11881596 TI - Foreign drug firm pleads guilty to felony charges. PMID- 11881597 TI - A 'touch' of diabetes? PMID- 11881598 TI - First contraceptive skin patch. PMID- 11881600 TI - FDA and the quality and integrity of research. PMID- 11881599 TI - Internet vendors warned about unapproved Cipro. PMID- 11881601 TI - FDA approves Viread for HIV-1 infection. PMID- 11881603 TI - Home male infertility test. PMID- 11881602 TI - Drug combination approved for advanced breast cancer. PMID- 11881604 TI - Study: aspirin and warfarin equally effective for stroke prevention. PMID- 11881605 TI - A smile away may help keep heart disease away. PMID- 11881606 TI - Study: treatment reduces risk of heart attack by 70 percent. PMID- 11881607 TI - Accutane risk management program strengthened. PMID- 11881608 TI - Teen science classes serve up lessons in food safety. PMID- 11881611 TI - Nipah virus. PMID- 11881612 TI - Nurses say health and safety concerns play a major role in employment decisions. PMID- 11881613 TI - New research confirms significant contribution made by multiple sclerosis specialist nurses. PMID- 11881614 TI - [New weapons against colorectal cancer]. PMID- 11881615 TI - Profiles of editorial board members. PMID- 11881616 TI - Immunobiology of liver dendritic cells. AB - Dendritic cells (DC) are rare, bone marrow-derived antigen-presenting cells that play a critical role in the induction and regulation of immune reactivity. In this article, we review the identification and characterization of liver DC, their ontogenic development, in vivo mobilization and population dynamics. In addition, we discuss the functions of DC isolated from liver tissue or celiac lymph, or propagated in vitro from liver-resident haemopoietic stem/progenitor cells. Evidence concerning the role of DC in viral hepatitis, liver tumours, autoimmune liver diseases, granulomatous inflammation and the outcome of liver transplantation is also discussed. PMID- 11881617 TI - [About regeneration and transplantation. 1926]. PMID- 11881618 TI - Needlestick injuries. PMID- 11881619 TI - Computer resources for nurses. PMID- 11881620 TI - Latex allergy. PMID- 11881622 TI - Management evaluation tool. PMID- 11881623 TI - Indicators of intimate partner violence in women's employment: implications for workplace action. AB - The study examined the indicators and consequences of intimate partner violence on women's employment and associated types and levels of violence. Interviews were conducted with a consecutive sample of 90 women seeking a protective order. Actual and threatened violence was measured with the Severity of Violence Against Women Scales (46 items). Eight questions were asked about type of harassment experienced at the worksite. The results found most of the abused women had been employed at one time (87%) and had also experienced harassment from an intimate partner related to their work (89%). Findings indicated lost productivity and reduced performance. The researchers concluded poor work performance, tardiness, and absenteeism may indicate an employee is suffering from intimate partner violence. PMID- 11881624 TI - Information technology: passport to the future. AB - Health care information in this millennium will become increasingly digital and electronically available. To keep pace and survive, occupational health leaders must determine the appropriate information technology strategy for their organization. The development and implementation of an electronic medical record can only be accomplished through a team effort that includes: management support to secure the necessary funding; participation of users to determine the application requirements and design; information systems expertise availability; and user education to ensure acceptance. The implementation of an electronic medical record is a journey, not a project, and it is the beginning of the organization's information infrastructure. Benefits include: increased staff efficiency with electronic versus paper folders; legible written documentation; multiple accessibility of medical records to authorized users; reduced potential for record loss/misfiling; and the ability to operate remotely and take advantage of more advanced technologies in the future. Occupational health staff must have the necessary computer applications on their desktops to increase their skills and enhance productivity. PMID- 11881625 TI - An effective transitional duty program: successful program. AB - The role of the employee health nurse practitioner/case manager in management of the Transitional Duty Program is pivotal to the program's success. Clearly articulated policies and procedures are used to support the Transitional Duty Program. Use of the transitional duty employee can produce financial benefits and improve employee morale. An effective Transitional Duty Program must successfully balance the needs of the employee with the needs of the organization. PMID- 11881626 TI - First lifetime back pain and psychiatry treatment: psychological factors and recovery in compensated workers. AB - The role of psychological factors in recovery from first lifetime low back pain (LBP) was explored in this study. Consecutive clients from one physiatry clinic in Montreal who had LBP of less than 3 months' duration, were on sick leave and receiving workers' compensation benefits, and reported the current event as first lifetime LBP were enrolled. Psychological factors that fluctuate with current events (Psychiatric Symptom Index) and remain stable over time (General Well Being Scale) were assessed. Outcomes were late return to work (> 31 days) and 1 year incidence of compensated recurrence. Results from two multivariate models indicated lower psychological distress predicted late return to work, and higher well being, higher aggressiveness, and lower anxiety predicted compensated recurrence. Researchers concluded psychological factors do not impact clients with all types of LBP in the same way. For individuals lacking prior LBP experience, better psychological functioning increased lengthy work absence. Thus, awareness of the clients' psychological profiles and previous LBP experiences may benefit recovery. PMID- 11881628 TI - Deborah Roy, Golden Pen Award winner. PMID- 11881627 TI - Confidentiality in occupational health care: a matter of advocacy. AB - Occupational and environmental health nurses play an important role in establishing and maintaining employee confidentiality. Preserving confidentiality transcends simple nondisclosure of private information and consists of an agreement between the employee and the health professional regarding the boundaries of shared secrets. Health professionals must be aware of specific limitations to confidentiality and convey these limits to employees. Occupational and environmental health nurses must restrict employer access to employee medical records. Open and honest communication is necessary to establish the appropriate boundaries of confidentiality and strengthen the employee/employer/health professional relationship. PMID- 11881629 TI - The association between generalized joint hypermobility and temporomandibular joint disorders: a systematic review. AB - To analyze conflicting evidence in the literature for the association between temporomandibular joint disorders (TMD) and generalized joint hypermobility (GJH), we performed a bibliographic search. The methodological quality of the 14 papers found was assessed according to 14 criteria. Papers were included in the analysis if the study population was clinically relevant, if range of motion of 2 or more joints was assessed on the left and right sides, and if cases had a TMD. Four studies fulfilled these selection criteria. Data from 3 studies, 113, cases and 95 controls, were available for analysis. Twenty-six cases and five controls were hypermobile (odds ratio, 5.4). In a sensitivity analysis, the odds ratio changed from significant to non-significant in 2 of 5 scenarios. As a result, it is not clear whether GJH is associated with TMD, and more rigorous studies are needed. PMID- 11881630 TI - The influence of various attachment types in mandibular implant-retained overdentures on maximum bite force and EMG. AB - The type of attachment that is used in oral rehabilitation by means of implant retained mandibular overdentures may influence the retention and the stability of the denture. In this study, we examined the hypothesis that a better retention and stability of the denture improve the oral function. Eighteen edentulous subjects received 2 permucosal implants, a new denture, and, successively, 3 suprastructure modalities. Maximum bite force and electrical activity of the masseter and temporalis muscles were measured. The maximum bite force nearly doubled after treatment for each of the 3 attachments. However, the average bite force after treatment was still only two-thirds of the value obtained for dentate subjects. No large differences in maximum bite force and muscle activity were found among the 3 attachment types. Temporalis activity was significantly lower than masseter activity when subjects clenched without implant support. There was no difference in activity when subjects clenched with implant support. PMID- 11881631 TI - Processing and properties of strong and non-rigid calcium phosphate cement. AB - A calcium phosphate cement (CPC) sets to form hydroxyapatite and has been used in dental and craniofacial applications. However, when CPC was used in periodontal repair, tooth mobility resulted in the fracture and exfoliation of the brittle implants. The aim of this study was to develop CPC-chitosan lactate composites with higher strength and increased strain before failure. It was hypothesized that the incorporation of chitosan lactate would render CPC non-rigid with improved properties. Two-way ANOVA showed significant effects of chitosan lactate and powder:liquid ratio (p < 0.001) on flexural strength, strain-at-peak-load, work-of-fracture, and elastic modulus. At powder:liquid = 2, the strength (mean +/- SD; n = 6) at 20% chitosan lactate was 15.7 +/- 1.3 MPa, higher than 4.9 +/- 1.4 MPa of CPC without chitosan lactate. At powder:liquid = 1, the strain-at-peak load was 0.2% for CPC without chitosan lactate; it increased to 15.8% for CPC containing 15% chitosan lactate. The work-of-fracture was increased by more than ten times. The novel strong and non-rigid CPC may provide compliance for tooth mobility without fracturing the implant, and may also extend the use of CPC into the repair of larger defects in stress-bearing locations. PMID- 11881632 TI - Issues in Amazonian development. PMID- 11881633 TI - Healthy discussion of planetary science goals. PMID- 11881634 TI - The nationality of a naturalist. PMID- 11881635 TI - First-time mothers' and fathers' transition to parenthood: infant care self efficacy, parenting satisfaction, and infant sex. AB - The threefold purposes of our study were to determine differences between first time mothers' and fathers' development of infant care self-efficacy and parenting satisfaction, relationships between mothers' and fathers' infant care self efficacy and parenting satisfaction, and the effect of infant sex on the development of mothers' and fathers' infant care self-efficacy and parenting satisfaction during the first 4 months following the infant's birth. A convenience sample of 44 couples in a midwestern state completed the Demographic Questionnaire, the Infant Care Survey, and What Being the Parent of a New Baby is Like-Revised. Fathers' reports of infant care self-efficacy increased linearly during the first 4 months following the infant's birth while mothers' reports of infant care self-efficacy increased linearly during the first 3 months. At all data collection times, fathers reported significantly lower infant care self efficacy than mothers. Reports of parenting satisfaction increased over time for mothers and fathers. At 8, 12, and 16 weeks following the infant's birth, mothers' infant care self-efficacy scores were significantly related to their parenting satisfaction scores. Fathers' infant care self-efficacy scores were significantly related to their parenting satisfaction scores at 12 and 16 weeks. Fathers of male infants had significantly higher parenting satisfaction scores than fathers of female infants at 12 and 16 weeks following the infant's birth. Nurses can develop individualized interventions to assist mothers and fathers during the transition to parenthood. PMID- 11881636 TI - [The heritage of hospitals and private philanthropy]. PMID- 11881637 TI - [The hospital of Belleville]. PMID- 11881638 TI - [Women serving the sick and the wounded in nineteenth century Russia]. AB - The genesis of women's service to the sick and the wounded dates from the 1820s. At the instigation of the tsarist court, assistance to the sick and wounded was provided by the so-called "widows of charity" recruited at the empress's command from among the inmates of an institution for poor officers' widows, then the Sisters of Charity. As early as the middle of the century, the Sisters' charity work had taken on a twofold character: during peacetime they cared for the poorest inhabitants, mainly of large and small towns; during wartime they cared for the sick and wounded in areas of armed conflict. From the time of the Crimean War, the Sisters of Charity constituted a permanent medical presence in all the wars of the Russian Empire. They were involved in the transportation of the sick and wounded, field hospitals as well as land and ship hospitals. Their activities later became institutionalised. In peacetime they served the poorest of the population, working in municipal hospitals, often founded near their convents since their organisation was modelled on Christian monastic structures. Three categories of Sisters of Charity were established: Orthodox, Catholic, though very small in number, and secular - among them the Russian Red Cross, from 1867, and their service to the sick and wounded has taken on the name of a "female profession". PMID- 11881639 TI - Genetic technologies and courts of law. AB - Human genetic testing, and its next logical technological plateau, gene therapy, are largely unregulated in the United States and much of the world. Since the basic science and some of the technological know-how related to such tests and therapeutic regimes cross national boundaries, the disputes that follow in the wake of popular response to such technological advances could arise in virtually any court of the world. For much of the post-industrial world, this pits the unfettered processes of medical decision-making against standards of the rule of law enforced by independent courts of law. For a substantial portion of the developing world, courts are not independent and the rule of law is precariously standardized. There the proponents of unbounded genetic test development and experimentation have greatest opportunities for mischief that can create civil, criminal and military strife. In this paper we propose and discuss methods to forecast case scenarios. We will emphasize the globalization of civil and criminal conflicts that may be spurred by the use of genetic testing. Extra judicial dispute resolution is likely to fail in the case of genetic tests. We finally propose an international means of judicial discourse that has potential to bring objectivity and even-handedness to the just resolution of issues spurred by genetic testing and engineered evolution. PMID- 11881640 TI - Quiz page. AA amyloidosis. PMID- 11881641 TI - Interaction of cosmic and solar flare radiations with the Martian atmosphere and their biological implications. AB - Assuming a constant interplanetary flux of galactic cosmic radiation and a model planetary atmosphere, it is possible to evaluate the magnitude of secondary ionization phenomena therein from parameters measured on Earth. The Martian atmosphere is of particular interest as its total air mass, estimated between 354 and 109 g cm-2, is in the vicinity of the Pfotzer cosmic ray maximum. Assuming the absence of a magnetic field on Mars the maximum neutron production would occur at an atmospheric depth of 75 +/- 5 g cm-2. With the lower air mass limit the surface flux of neutrons reaching the Martian surface could be about 240 times greater than observed at Earth's sea level. Surface minerals containing nuclei with large capture cross sections for slow neutrons, such as Li6, B10 and U235, could thus serve as valuable indicators for the age of the Martian crust. In general, the tenuous Martian atmosphere would result in greater surface radiation dose rates, particularly during times of relativistic solar flares. If the surface air mass is as low as 109 g cm-2 then the rate of nuclear disintegrations due to galactic cosmic radiation would exceed that on Earth's sea level approximately 1000-fold. The tenuous Martian atmosphere would not be a complete shield for heavy primary nuclei and about 1 percent of the incident flux could reach the surface. PMID- 11881642 TI - Bacteria under simulated Martian conditions. AB - The behavior of organisms in simulated Martian conditions is of great importance to exobiology for two reasons: (1) Because of the extreme environment of Mars, the likelihood of contamination of the planet by earth organisms is considered slight by some scientists. To date, there has been little evidence to contradict this supposition. Such evidence is presented. (2) The selection and adaptation of earth bacteria to Martian conditions is potentially significant in understanding Martian life, if it exists, and may be helpful in designing life-detection techniques and devices. Of course, simulation attempts, based on current knowledge of the Mars environment, may be far from the actual conditions, and extrapolations made from such situations of no real significance. However, generalizations can be made and cautious interpretation of the results of those experiments seems well worth reporting. A new technique for simulation of known parameters of the Martian environment is discussed along with possible biological implications. The response of bacteria to such simulation is demonstrated in terms of survival and growth, showing that certain bacteria will not only survive, but grow during simulated Martian freeze-thaw cycling if water is present. Ways are demonstrated in which water can be present on Mars although not detectable with current technology. Plans for future experimentation are discussed. PMID- 11881643 TI - "Gulliver", an experiment for extraterrestrial life detection and analysis. AB - Based on the probability that extraterrestrial life is biochemically somewhat similar to life on Earth, a life detection experiment is being prepared to explore Mars. The experiment will be performed by an automated device which will carry a microbiological medium being developed to support a wide range of earth microorganisms. Selected ingredients of the medium will be labeled with radioactive isotopes. A sticky string, shot out from and reeled back into the device, will gather a sample of the Martian soil. It is hoped the radioactive atoms in the compounds will be metabolized by the unknown organisms in the soil and evolved in a labelled gas. The gas will be collected by a chemical "getter" and the radioactivity measured for transmission to Earth. A positive response from the test unit and a negative, or lesser, response from a poisoned control unit would constitute evidence of life. The device can also differentiate between photosynthetic and nonphotosynthetic metabolic activity. Data from field tests on Earth are presented. PMID- 11881644 TI - Life and space. AB - The studies dealing with the detection of life on planets and in space will have two periods. The first period will be associated with the application of automatical devices, which will detect living creatures, whereas the second, more late, period will be concerned with the investigations provided by man himself. In these studies one should proceed from the idea of life on the Earth, and, consequently, look for living creatures containing no water, no carbon dioxide, etc. only after obtaining negative results in searching for creatures similar to those living on the Earth. The whole course of "chemical evolution" on the Earth proves the necessity of the detection on other planets of heterotrophic microorganisms at first, and only afterwards to look for specialized forms, i.e. chemo- and photoautotrophs. It is usually underestimated that as a result of adaptation to certain ecological conditions existing on planets living creatures may appear, which will distinctly differ in their biological properties from terrestrial forms. At present there is no faultless investigation which is able to prove to existence of cosmobionts in meteorites. The appropriate experiments convince us that soil bacteria. may penetrate into the central parts of meteorites, which have been lying in the ground. Samplings at high altitudes with the aid of aerostats or the rockets are difficult to provide because of the possibility of pollution of samples by terrestrial microbes. It is therefore necessary to elaborate special devices and new methods for sampling. The most perfect instrument for the detection of microorganisms will be a device, which will record the multiplication rate of microbes in the liquid media simultaneously by means of different methods: manometry, nephelometry, potentiometry and the determination of radioactive carbon dioxide which is extracted during the breakdown of organic substances. PMID- 11881645 TI - The effect of low temperatures on the structure of enzymes. AB - The enzymes of the cold stable organisms possess lower levels of activation energy. A possibility for the adaptation of living creatures to low temperatures is finally associated with the structural alterations in proteins and enzymes. Polarographic studies on yeast alcohol dehydrogenase and on proteinase of B. subtilis at low temperatures have demonstrated that a lowering of temperature causes significant structural changes in the protein molecule. PMID- 11881646 TI - Effects of gravity on the functions of the central nervous system. AB - The sensitivity of the mammalian central nervous system to gravitational influences involves both direct and indirect factors. Gradual loss of cerebral circulation with increasing acceleration beyond 5 G has been shown to evoke changes in patterns of brain electrical activity, with epileptiform discharges triggered primarily in the hippocampal system of the temporal lobe, and spreading into other brain systems. The relationship of these structures to judgment and discriminative functions is discussed. Complete loss of cerebral circulation is associated with electrical silence in the brain. The possible effects of weightlessness on intracranial fluid distribution are reviewed. Tests of vibrational stimuli in the monkey have disclosed "driving" of electrical brain rhythms at the shaking frequency, particularly in the range from 11 to 15 cycles per second. These effects are unrelated to photic stimulation, and are abolished by anesthesia or death. Tests of discriminative capability at frequencies producing maximum driving have shown increased response latencies and increased errors. The neurophysiological basis for adaptive phenomena in recurrent vestibular stimulation has been found to reside partly in the vestibular nuclei of the medulla, and not to require integrity of connections with higher vestibular centers. The potentiation of responsiveness to vestibular stimulation in the weightless state has been shown to have a basis in the exaggerated responsiveness of medullary vestibular centers following loss of proprioceptive influxes from the spinal cord. The possible contribution of weightlessness to anomalous psychophysiological functions is reviewed. This area presents a major challenge in experimental design, and may require evaluation of such phenomena as distortion of the body image, modified sleep patterns and changes in optimal sleep-work cycles. PMID- 11881647 TI - Cardiovascular deconditioning: role of blood volume and sympathetic neurohormones. AB - Studies of prolonged bed rest and water immersion have amply demonstrated that serious deconditioning of certain gravity-oriented biologic systems occurs with extended exposures to environments in which the necessity for gravity compensation has been reduced. The precise physiologic mechanisms responsible for the loss of orthostatic tolerance following such studies are unknown, but may be related to diminished blood volume, decreased muscle or tissue pressure in the extremities or to functional alterations in the sympathetic nervous system. In view of the fact that noradrenaline is the principal agent for maintaining vascular tone the author has focused his attention on the sympathetic nervous system as being the system most likely responsible for the loss of orthostatic tolerance following such experiments. The contributory effects of diminished circulating blood volume resulting from such environments are also discussed. PMID- 11881648 TI - Calcium metabolism under conditions of weightlessness. AB - This is a brief report on the possible effects of weightlessness on calcium metabolism. Interest in this topic is based on common, clinical experience: immobilization from any cause is accompanied by skeletal rarefaction and hypercalcuria. PMID- 11881651 TI - Effect of ultraviolet radiation on microorganisms as a principal extremal factor of space environment. AB - 1. The problem of the transport of viable germs through interplanetary space has recently acquired special interest arising from the necessity of sterilization of space ships. 2. Of all cosmic extremal factors ultraviolet solar radiation is the most dangerous for microorganisms. The intensity of the most bactericidal region of ultraviolet radiation at a distance of one astronomical unit from the sun is 2 x 10(3) erg per cm2. 3. Sensitivity of microorganisms to ultraviolet rays varies in a large range. For the most resistant forms the lethal dose is approximately 440000 erg per cm2. 4. The penetrating capacity of ultraviolet radiation is very low, and therefore, even insignificant amounts of organic or mineral substances may protect the bacterial cell. 5. Dust particles of terrestrial origin carrying on themselves bacterial spores exert upon the latter a protective effect against ultraviolet radiation. PMID- 11881649 TI - The atmospheres of Mars, Venus and Jupiter. AB - The general properties of the atmospheres of Mars, Venus and Jupiter are reviewed. The evolutionary processes of planetary atmospheres are outlined as an introduction to a detailed discussion of the structure of the atmospheres of other planets. The most recent observational results indicate that the surface pressure on Mars could be as low as 20 mb, and the amount of CO2 may have a mixing ratio as high as 25% by volume. For Venus, the Mariner II results suggest that the surface temperature may actually be of the order of 600 degrees K. Also there are indications that the surface pressure on Venus could be approximately 100 atm., with the atmosphere mainly composed of N2. In the case of Jupiter, recent theoretical investigations of the atmosphere below the clouds suggest, that there may be regions in the atmosphere of very high density and pressure but at relatively low temperature, creating conditions possibly favorable to the occurrence of complex organic reactions. PMID- 11881650 TI - Life sciences and space research II: a session of The Fourth International Space Science Symposium, Warsaw, June 3-12, 1963. PMID- 11881653 TI - The vestibular function in conditions of zero gravity. AB - In the pigeon vestibular responses from threshold to saturation will be compared in laboratory and in a satellite zero gravity. It is expected that following the Weber-Flechner law sensitivity in zero gravity conditions will be much higher. The experiment in orbit and the necessary equipment are described. PMID- 11881652 TI - The action of magnetic field on the sodium transport across the cell membrane. AB - In the frog skin Na+ transport has been studied using 24Na+ and the short circuit technique, with or without a 650 gauss steady magnetic field. The magnetic field reduced the influx of sodium by 10-30%. In short circuit experiments the current appeared to be altered within 1 second of increasing the field. Such effect seems to be due to a block in active transport of sodium since the addition of 3 mM ouabain to the inside solution prevented the change. PMID- 11881654 TI - Use of algae for support of the human in space. AB - Algae offer special advantages as photosynthetic plants which may be used to provide for the gas exchange requirements of man and for some fraction of his food requirement. Established principles of algal metabolism allow basic specifications. To meet a human oxygen demand of 600 1/day an algal exchanger would take up about 720 1/day of carbon dioxide, produce about 600 g/day of dry algae, and require a minimum of about 800 watt of visible light. Major technical disadvantage arises from need for input power as light and the low values of irradiance at which algae can maintain maximum efficiency. Design criteria require compromise between minimum volume and minimum power requirement. A basic design theory needs to be evolved and a number of technical problems need to be solved before proper evaluation of an algal system can be made. PMID- 11881655 TI - Exobiology: a critical review. AB - The quest for life beyond the Earth can be implemented in three ways. First, we may study contemporary terrestrial biology and organic chemistry to approach the problem of the origin of life on Earth. If it appears relatively easy for life to have emerged in the primitive terrestrial environment, it may follow that the origin of life is a fairly general planetary phenomenon. Second, we can investigate the physical environments of the planets, and determine whether the conditions there are so severe that living processes are entirely excluded, even allowing for the adaptability of life. Third, the planets may be studied for direct evidence of indigenous life forms. All three approaches are beset with uncertainties, and it is important to state at the outset that no completely convincing evidence exists for extraterrestrial life. The problem often reduces to probability considerations, and to estimates of observational reliability. At convenient places in the marshalling of evidence, I shall try to pause and give brief expression to alternative interpretations. In almost all cases, an optimistic view can be found which holds that the evidence is strongly suggestive of, or at the worst, not inconsistent with, the existence of extraterrestrial life; and a pessimistic view can be found, which holds that the evidence adduced in favor of extraterrestrial life is unconvincing, irrelevant, or has an alternative, non-biological explanation. I leave it to the reader to pick his own way among the factions. PMID- 11881656 TI - Organic constituents of carbonaceous chondrites. AB - From a brief discussion of forms of meteorite carbon it is concluded that almost all the carbon in the carbonaceous chondrites is present as organic matter. Attempts to extract and identify this organic matter are then reviewed. It is shown that only 25 per cent has been extracted and only about 5 per cent chemically characterized. Of this 5 per cent, most is a complex mixture of hydroxylated aromatic acids together with various hydrocarbons of the paraffin, naphthene and aromatic series. Small amounts of amino acids, sugars and fatty acids also are present. The possible chemical nature of the major fraction is discussed. It is suggested to be a mixture of high-molecular weight aromatic and hydrocarbon polymers. Possible sources of contamination of the meteorites are described and evidence indicating a general lack of organic contaminants is presented. It is concluded, that most of the organic constituents are indigenous to the meteorites and are extra terrestrial in origin. Synthetic processes for the compounds are mentioned and it is concluded that the organic material is probably of abiogenic origin. A brief review on studies of "organized elements" contained within the meteorites is presented. Difficulties of identification are discussed and photographs of some microstructures of several carbonaceous chondrites are presented. No final conclusion about the nature of these objects is possible, but some appear to be various indigenous organic and mineral structures, while others are terrestrial contaminants. PMID- 11881657 TI - Reflection spectra of bio-organic materials in the 2.5-4 micron region and the interpretation of the infrared spectrum of Mars. AB - The nature and assignment of the Martian spectral features observed by W. M. Sinton in the 3-4 micron region have been re-examined. It is shown that it is not possible to state unequivocally that there are absorption bands at 2710 (3.69 microns), 2790 (3.58 microns) and 2900 (3.45 microns) cm-1. Rather the possibility of the presence of true reflection features complicates the issue and enables only a wide range to be specified for the corresponding resonant frequencies. To clarify the assignment of these a large number of pure organic and inorganic solids, together with a selection of biological samples, have been recorded in reflection. The assignment of the features to organic matter, implying the presence of life on the Martian maria, is found to be improbable. While inorganic carbonates have bands in this region, they also do not provide a satisfactory explanation of the observations. The assignment of these Martian spectral features is then an open question. PMID- 11881658 TI - Terminal sedation--a possible compromise in the euthanasia debate? PMID- 11881659 TI - New developments in human genetics. Statement of a WHO expert consultation. PMID- 11881660 TI - Whistleblowing heroes--boon or burden? PMID- 11881661 TI - Guidelines for obstetric anaesthetic research. AB - Published in May this year, after consultation with a variety of relevant groups including appropriate consumer organizations, these guidelines are designed both to safeguard the interests of research participants and to facilitate the research process. PMID- 11881662 TI - Another look at LREC annual reports. AB - Almost the only readily available sources of information about the activities of UK research ethics committees are the annual reports that they have been required since 1991 to publish. This paper reports on a further study of those reports by the Bulletin. PMID- 11881663 TI - Science versus man? Meeting held in Massa and Carrara, Italy, 28-30 September 2001. PMID- 11881664 TI - The promotion of scientific medical research at the service of man from the point of view of cultural effectiveness. PMID- 11881665 TI - Morality and economics in health care. PMID- 11881666 TI - Other religions and economy and health: reflections from conciliar Protestantism. PMID- 11881667 TI - The Jewish view of the economy and health. PMID- 11881668 TI - The economy and health in the light of Islamic principles. PMID- 11881669 TI - The economy and health: the Hindu perspective. PMID- 11881670 TI - The road to abortion (II): how government got hooked. AB - The first part of this series traced close links between eugenics (the effort to breed a "better" human race) and population control throughout the greater part of this century up to the 1960s. It stressed the population work of early eugenicists and eugenics sympathizers such as Frederick Osborn, Margaret Sanger, Gunnar Myrdal, Alan Guttmacher, Garrett Hardin and John D. Rockefeller 3rd. This second and concluding part will show how population controllers, from the 60s onward increasingly added economic and foreign-policy concerns to their original "eugenics" motive of improving human genetic stock. Working in both Democratic and Republican administrations, they gained major government backing for their programs and also played a key role in the legalization of abortion. I will use President Richard Nixon's administration as an example of heavy government involvement. PMID- 11881671 TI - Starved to death by order of the court. PMID- 11881677 TI - Kids and euthanasia. PMID- 11881686 TI - Human genetics commission launches consultation on the future of genetic information. PMID- 11881687 TI - Protocol banning human cloning entered into force on the list of March 2001. PMID- 11881688 TI - Legal responses to some of the new developments in reproductive technologies. Part 1. PMID- 11881689 TI - Ethical, biological and legal aspects in the use of human embryonic stem cells in Germany. PMID- 11881691 TI - A discussion of social, ethical, practical and consequential effects of sex selection. PMID- 11881690 TI - 'Not so wrong that we are prepared to threaten the entire service': the regulation of reproductive technologies in the UK. AB - In the UK human embryo research and infertility treatments are regulated by the Human Fertilisation and Embryology Authority (HFEA), a national statutory body set up in 1991. Clinics are required by law to be licensed by the HFEA and open to inspection. In 1999 there were 107 clinics licensed to carry out IVF and/or donor insemination. There are regional variations in National Health Service funding and seventy-five percent of those treated pay for most of their treatment, usually to avoid long waiting lists or because they do not meet the eligibility criteria set by their local health authority. Having described the code of practice laid down by the HFEA the paper discusses the implications of new reproductive technologies from a social science perspective, focusing on three topics which continue to be debated by the media, the regulatory authorities, health professionals and 'lay' people. First, age limits and other criteria for assisted reproduction which raise the question 'who should be allowed to reproduce?' Second, donor anonymity and the question of payment for donors. Finally, the screening and selection of embryos for implantation and the recurring theme in the media of 'designer babies'. The HFEA can be seen to fulfill its intended role, mediating between the infertility professionals, patients and the wider general public. It seeks to allay public fears while supporting professionals by avoiding restrictions on research and treatment and by consulting with the public as to when extensions to research will be acceptable. PMID- 11881692 TI - Two notable pioneers in conscious sedation pass their gifts of pain-free dentistry to another generation. PMID- 11881693 TI - Oral transmucosal fentanyl pretreatment for outpatient general anesthesia. AB - The oral transmucosal formulation of fentanyl citrate (OTFC) has been reported to be an effective sedative, providing convenient and atraumatic sedation for children prior to general anesthesia or painful diagnostic procedures. Thirty three young children (24-60 months of age) scheduled for outpatient general anesthesia for treatment of dental caries were enrolled in this randomized placebo-controlled clinical trial. To determine the effectiveness of the OTFC premedication, patient behavior was evaluated using three distinct outcome ratings. A sedation score rated behavior in the waiting room prior to OTFC as well as 10 minutes and 20 minutes after OTFC. A separation score rated the child's response to being separated from his/her parent or guardian for transport to the dental operatory. Finally, a cooperation score rated the child's acceptance of the mask induction. The OTFC formulation was well tolerated by most of the children in this study. Compared with the placebo oralet, the active OTFC improved behavior for separation from the parent (P < .05) and cooperation with the mask induction (P < .05). The duration of surgery and the time of recovery did not differ between placebo and active premedication. Side effects including respiratory and cardiovascular complications were reported more frequently in the active fentanyl group. Continuous monitoring of respiratory function is essential when using this unique and effective formulation of fentanyl for pediatric preanesthetic sedation. PMID- 11881694 TI - Comparative study on anesthetic potency of dental local anesthetics assessed by the jaw-opening reflex in rabbits. AB - The potency of 4 local anesthetics to dental pulp was compared. Drugs were 4% articaine with 12 microgram/mL epinephrine (A12), 4% articaine with 6 microgram/mL epinephrine (A6), 2% lidocaine with 12.5 microgram/mL epinephrine (L), and 3% propitocaine with 0.03 IU/mL felypressin (P). Local anesthetics were injected into the dental root of the mandibular incisor. Electromyogram (EMG) of the digastric muscle was measured during the jaw-opening reflex induced by electrical stimulation. The disappearance of the EMG wave was judged as positive evidence of anesthesia. The determination of ED50 of the anesthetic was made by probit analysis. The ED50 of the A12 was minimal in all the tested anesthetics throughout the entire course. The potency in the A6 was 2.8 times that of the L. The potency of the A12 at the 15-minute measurement was 3.8 times that of the A6. The ED50 of the P was higher compared with those of the other 3 groups. It was concluded that articaine showed quicker onset than lidocaine and propitocaine and that there was a need to increase the dosage to attain a quick onset or to extend the duration. PMID- 11881695 TI - Clinical development units. PMID- 11881696 TI - Effects of performance feedback on patient pain outcomes. AB - The purpose of this study was to examine the effects of an individual feedback intervention provided to nurses on selected patient outcomes related to postoperative pain management Individual performance feedback served as the intervention. Thirty orthopedic staff nurses received feedback information on their past performance of recommended pain management practices. Data were collected preintervention and postintervention on selected patient pain outcomes from the medical records of 240 patients who had undergone total knee arthroplasty. The patient outcome measures were mean 4-hour pain intensity ratings, mean highest pain intensity ratings, mean number of times pain ratings exceeded patients' acceptable level of pain, mean administered morphine equivalents, and mean pain ratings on reassessment following analgesia administration. An improvement was noted in all pain outcomes following the feedback intervention. Results indicate that providing nurses with feedback on their past performance of pain management practices may contribute to decreased postoperative pain. PMID- 11881697 TI - Developmental evolution of expertise in diabetes self-management. AB - The following is a description of the findings of a longitudinal exploratory and descriptive research study of 22 persons nominated as expert self-managers of Type 1 diabetes. It entailed an initial interview about previous experiences with self-management, self-recorded taped diaries about self-management decisions for 1 week each, and face-to-face interviews following each weeklong recording of self-management decisions. The study generated a grounded theory about the development of expertise in diabetes self-management. The development of expertise was found to occur as transition through two or more phases, to be individualized, and to involve a complex interplay between social, contextual and personal factors, including the individual's developmental age. The research fIndings challenge the traditional understanding of rebellion in self-management as a manifestation of adolescence, behaviors other than active control as testimony to ineptitude in self-management, metabolic control as the indicator of self-management ability, and the role of others as collaborators in self management. PMID- 11881698 TI - Cognitive function in patients with Cushing syndrome: a longitudinal perspective. AB - The purposes of this study were to examine the level of improvement of cognitive function 12 months posttreatment in adult patients with Cushing syndrome (CS), the relationships of cognitive function to duration of CS or recovery of the hypothalamic pituitary adrenal (HPA) axis, and depression and improved cognitive functioning. Thirty-three patients with CS and a matched comparison group were enrolled. IQ, depression, and endocrine factors were measured during the active phase of CS and at 12 months posttreatment for CS. Results show no group differences in cognitive function across time but a trend for CS patients to have lower IQ scores at baseline. Individual differences in performance were striking. For some subscales of IQ there was a positive relationship with recovery of the HPA axis and a negative relationship with duration of CS as well as an improvement if depression had decreased. Limitations of the study are cited along with clinical implications and directions for future research. PMID- 11881699 TI - Nurse comforting strategies: perceptions of emergency department patients. AB - Throughout nursing's history, comfort has been a desired outcome in the provision of nursing care. Yet, attempts to understand comfort and comforting have only recently emerged. To date,few studies have explored comfort and comforting from patients' perspectives. The purpose of this qualitative exploratory study was to describe nurse comforting strategies as well as the outcomes of nurse comforting strategies from the perspective of emergency department (ED) patients. The study took place in a rural Canadian acute care regional hospital. A volunteer sample of 14 hospitalized patients who had received initial treatment in the ED were interviewed. Interviews were transcribed and content analysis performed. Participants described nurses' comforting strategies under the following categories: immediate and competent technical/physical care, positive talk, vigilance, attending to physical discomforts, and including and attending to family. The comforting strategies used by nurses had a positive impact on the physical and emotional well-being of the participants. PMID- 11881700 TI - Facilitating and constraining factors on autonomy: the views of stroke patients on admission into nursing homes. AB - This article describes a model changing autonomy which was developed in a grounded theory study among stroke patients on admission into nursing homes for rehabilitation. Three dimensions of autonomy were identified: self-determination, independence, and self-care. On admission, patients' conditions (disabilities, multimorbidity, emotional state, and feeling like a layperson) and patients' strategies (waiting and seeing, and acting as a subordinate) constrain autonomy. Several environmental factors facilitate patient autonomy. The nursing home sustains patient autonomy by providing a hopeful atmosphere and room for autonomy. The health professionals facilitate autonomy by giving therapy, support and information, attentiveness and respect, paternalism and teamwork, Facilitating strategies of the family encompass emotional and instrumental support Care routines, lack of privacy, an unfamiliar environment, waiting periods, boredom, and lack of information were identified as constraining environmental factors. Developing guidelines and multidisciplinary courses regarding the approach to patient autonomy on admission is recommended. PMID- 11881701 TI - Comparison of different methods of obtaining orthostatic vital signs. AB - The purpose of this study was to compare two lying and standing procedures for measuring orthostatic vital signs. Thirty-five normotensive participants (mean age 21.6 years)participated in a randomized crossover study. Measures of blood pressure (BP), heart rate, and dizziness were collected at different lying and standing times. AU subjects participated in a standardized walk paced at 4 miles per hour prior to lying. Using analysis of variance (ANOVA) with post hoc contrasts, the mean systolic BP differed between 5 and 10 minutes of lying (F = 21.33, p < .001) and the mean diastolic BP tended to differ between those time points (F = 5.23, p < .03). The mean standing systolic BP and dizziness rating were different between 0- and 2-minute intervals (F = 8.36, p < .01 and F = 7.15, p < .10). In normotensive participants following standardized exercise, orthostatic vital signs stabilized after lying 10 minutes. PMID- 11881702 TI - Evaluation of select variables in the ion chromatographic determination of F-, Cl , Br-, NO(-)3, SO(-2)4, and PO(-3)4 in serum samples. AB - A full experimental design at two levels is applied for the estimation of the significance of select factors that may influence the ion chromatography (IC) determination of F-, Cl-, Br-, NO(-)3, SO(-2)4, and PO(-3)4 in serum samples. The factors studied are various sample deproteinization procedures, eluent composition, and flow rates. Deproteinization using either acetonitrile-NaOH or ultrafiltration can be used in order to obtain a significant protein removal before IC analysis; however, the former is recommended because it is less time consuming and cheaper. Better resolution is obtained when a sodium hydroxide solution is used as the eluent. There is no influence of the sample's deproteinization procedures on the chromatographic resolution. PMID- 11881703 TI - Temperature effect on peak width and column efficiency in subcritical water chromatography. AB - Subcritical water has been recently employed as the mobile phase to eliminate the use of organic solvents in reversed-phase liquid chromatography. Although the influence of temperature on retention in subcritical water chromatography has been reported, the temperature effect on peak width and column efficiency has not yet been quantitatively studied. In this work, several polar and chlorinated compounds are separated using pure subcritical water on Zorbax RX-C8, PRP-1 (polystyrene-divinylbenzene), Hypersil ODS, and ZirChrom-polybutadiene columns. Isothermal separations are performed at temperatures ranging from 60 degrees C to 160 degrees C. The retention time and peak width of analytes are reduced with increasing temperature. However, the column efficiency is either improved or almost unchanged with the increasing temperature in the low-temperature range (lower than the 100 degrees C to 120 degrees C range), but it is decreased when temperature is further raised in the high-temperature range (higher than the 100 degrees C to 120 degrees C range). Therefore, a maximum in column efficiency is obtained at temperatures within the 100 degrees C to 120 degrees C range in most cases. PMID- 11881705 TI - Quantitative variability. PMID- 11881704 TI - A high-pressure liquid chromatographic-tandem mass spectrometric method for the determination of ethambutol in human plasma, bronchoalveolar lavage fluid, and alveolar cells. AB - A technique is presented for the specific and sensitive determination of ethambutol concentrations in plasma, bronchoalveolar lavage (BAL), and alveolar cells (AC) using a high-pressure liquid chromatographic (HPLC)-tandem mass spectrometric (MS-MS) method. The preparation of samples requires a deproteinization step with acetonitrile. The retention times for ethambutol, neostigmine bromide, and propranolol are 2.0, 1.4, and 1.1 min, respectively, with a total run time of 2.8 min. The detection limits for ethambutol are 0.05 microg/mL for plasma and 0.005 microg/mL for the BAL supernatants and AC suspensions. The assay has excellent performance characteristics and has been used to support a study of the intrapulmonary pharmacokinetics of ethambutol in human subjects. PMID- 11881706 TI - A study of polyethoxylated alkylphenols by packed column supercritical fluid chromatography. AB - Alkylphenol polyethoxylates (APEs) are a widely used group of nonionic surfactants in commercial production. Characterization of the composition of APE mixtures can be exploited for the determination of their most effective uses. In this study sample mixtures contain nonylphenol polyethoxylates and octylphenol polyethoxylates. The separation of individual alkylphenols by ethoxylate units is performed by supercritical fluid chromatography (SFC)-UV as well as normal-phase high-performance liquid chromatographic (HPLC)-UV employing packed columns. The stationary phase and column length are varied in the SFC setup to produce the most favorable separation conditions. Additionally, combinations of packed columns of different stationary phases are tested. The combination of a diol and a cyano column is found to produce optimal results. An advantage of using packed columns instead of capillary columns is the ability to inject large amounts of sample and thus collect eluted fractions. In this regard, fractions from SFC runs are collected and analyzed by flow injection analysis-electrospray ionization mass spectroscopy in order to positively identify the composition of the fractions. In comparing the separation of APE mixtures by SFC and HPLC, it is found that SFC provides shorter retention times with similar resolution. In addition, less solvent waste is produced using SFC. PMID- 11881707 TI - An isocratic liquid chromatographic method with diode-array detection for the simultaneous determination of alpha-tocopherol, retinol, and five carotenoids in human serum. AB - An isocratic high-performance liquid chromatography (HPLC) method for the simultaneous determination of alpha-tocopherol, retinol, and five carotenoids (lutein-zeaxanthin, beta-cryptoxanthin, lycopene, and alpha- and beta-carotene) in human serum is described. Serum samples are deproteinized with ethanol and extracted once with n-hexane. Resulting extracts are injected onto a C18 reversed phase column eluted with methanol-acetonitrile-tetrahydrofuran (75:20:5, v/v/v), and full elution of all the analytes is realized isocratically within 20 min. The detection is operated using three channels of a diode-array spectrophotometer at 290, 325, and 450 nm for tocopherol, retinol, and the carotenoids, respectively. An internal standard is used for each channel, which improves precision. The choice of internal standards is discussed, as well as the extraction protocol and the need for adding an antioxidant during the extraction and chromatographic steps. The analytical recoveries for liposoluble vitamins and carotenoids are more than 85%. Intra-assay relative standard deviation (RSD) values (n = 20) for measured concentrations in serum range from 3.3% (retinol) to 9.5% (lycopene), and interassay RSDs (n = 5) range from 3.8% (alpha-tocopherol) to 13.7% (beta cryptoxanthin). The present method is used to quantitate the cited vitamins in healthy subjects (n = 168) from ages 9 to 55 years old. PMID- 11881708 TI - A procedure for sampling and analysis of air for energetics and related compounds. AB - A procedure for the sampling and analysis of energetics and related compounds in the atmosphere is described. The basic procedure consists of the collection of air samples using sampling cartridges containing XAD-2 resin, extraction of the resin with isoamyl acetate, and an analysis of the extract using gas chromatography with electron capture detection. Modifications and additions to this procedure are discussed, such as the use of a prefilter before the resin sampler to collect particulates and the use of a mass selective detector to analyze for some propellant compounds of interest or for quantitative confirmation purposes. Two differing sizes of samplers are evaluated according to the air volumes required for collection. The procedure is tested through the analysis of spiked resin samples, which had air pulled through them for periods of time corresponding with the required sampling volumes. This procedure has application toward the measurement of energetic residues in atmospheres resulting from weapons testing and operations during training exercises involving munitions. PMID- 11881709 TI - Displacement study on a vancomycin-based stationary phase using N-acetyl-D alanine as a competing agent. AB - The analysis of the binding data of D,L-dansyl amino acids on a vancomycin stationary phase is investigated in relation to the addition of N-acetyl-D alanine in the mobile phase. This eluent additive acts as a specific competing agent for the aglycone pocket of the immobilized chiral selector. A model taking into account both stereoselective and nonstereoselective interactions between the solutes and the stationary phase is used to fit the experimental data. From the results, the theoretical approach is considered to be adequate to describe the competing agent dependence on solute retention. To the best of our knowledge, this report constitutes the first example of a displacement study on a macrocyclic antibiotic stationary phase. This work shows that dansyl amino acids bind to the active aglycone pocket of the selector and that this interaction is enantioselective. The results also demonstrate that additional enantioselective sites at the vancomycin surface are involved in the chiral discrimination of these solutes. PMID- 11881710 TI - Influence of hydrolysis, purification, and calibration method on furosine determination using Ion-pair reversed-phase high-performance liquid chromatography. AB - The influence of HCI concentration (6M, 8M, and 10M) and the ratio of sample protein to acid (1 or 5 mg of protein per mL of acid) on furosine formation during sample hydrolysis is studied. The conditions that maximize furosine formation are 10M HCI in the ratio of 1 mg of protein to 1 mL of acid. Purification of the hydrolysate by solid-phase extraction is also considered by examining the effect of hydrolysate volume and volume of 3M HCI used to elute the furosine. Furosine quantitation is carried out using the standard additions and external standard methods. The results indicate that there is no interference by the sample matrix and that external calibration is adequate. PMID- 11881711 TI - The use of nonendcapped C18 columns in the cleanup of clenbuterol and a new adrenergic agonist from bovine liver by gas chromatography-tandem mass spectrometry analysis. AB - More specific official methodology is needed to survey the illegal use of clenbuterol in animal production plus the synthesis of new compounds that currently elude routine analytical methods. The identification of a new adrenergic agonist, N1-(2-(4-amino-3,5-dichlorophenyl)-2-hydroxyethyl)-N1 isopropyl-propanamide (known as compound A) in animal feed has prompted studies to verify if the existing cleanup procedures developed for clenbuterol are really effective. This study considers the ion-exchange mechanism on cyanopropyl (CN), sulfonic cation exchange (SCX), mixed phase (MPH) (C8 + SCX), and nonendcapped C18 (C18NE) solid-phase extraction (SPE) columns. Results indicate that compound A (by contrast with clenbuterol) is not efficiently retained on the CN, SCX, and MPH SPE columns (recovery <10%). This finding thus leads to the development of a gas chromatography-tandem mass spectrometry procedure based on C18NE SPE that is able to purify both agonists from bovine livers spiked at 0.5, 1.0, and 2.0 ppb with a mean recovery of 93% for clenbuterol and 92% for compound A. PMID- 11881712 TI - Simultaneous high-performance liquid chromatographic determination of paracetamol, phenylephrine HCl, and chlorpheniramine maleate in pharmaceutical dosage forms. AB - A rapid, precise, and specific high-performance liquid chromatographic method is described for the simultaneous determination of paracetamol, phenylephrine HCI, and chlorpheniramine maleate in combined pharmaceutical dosage forms. The method involves the use of a microBondapak CN RP analytical column (125 A, 10 microm, 3.9 x 150 mm) at 22 degrees C as the stationary phase with the mixture of acetonitrile and phosphate buffer (pH 6.22, 78:22) as the mobile phase. Derivatization of the drugs is not required. The method is applied to commercial pediatric cough-cold syrups, tablets, and capsules marketed in Turkey. The relative standard deviation for 10 replicate measurements of each drug in the medicaments is always less than 2%. PMID- 11881714 TI - Generalizability and clinical utility of a practice-based infant pain assessment instrument. AB - The purpose of this study was to determine the clinical usefulness and generalizability of an infant pain assessment instrument. Earlier work showed that this instrument--an algorithm derived from a model of infant pain assessment possessed excellent content validity, criterion-like validity, and 3-month stability (test-retest reliability). In this study, generalizability was determined by comparing the percentage agreement between inexperienced pediatric nurses and one author, both using the tool to assess pain of infants in various clinical settings, and by comparing the percentage agreement between one author who used the tool to assess pain and the infant's pediatric nurse caretaker who used his or her clinical expertise, not the tool, to assess pain across various clinical settings. The results show excellent generalizability. PMID- 11881713 TI - Assessing infant health promotion: a cross-cultural comparison. AB - The purpose of this cross-cultural, correlational study was to evaluate two popular clinical/research assessment tools, the NCAST Teaching Scale and the Home Observation Measurement of the Environment (HOME) Inventory, as measures of infant health promotion behaviors for low-income, foreign-born Hispanic mothers in the United States. Based on the assumption that both measures tap universal attributes of the mother-infant relationship, it was hypothesized that maternal performance for the study group and a comparison group of U.S.-born, low-income mothers would be similar. Comparable performance on the NCAST Teaching Scale included a full range of scores, including the capacity to identify mothers most in need of clinical intervention. Study fundings supported the clinical use of this assessment scale with mothers represented by the study sample. Consistently lower scores by the foreign-born Hispanic mothers on the HOME Inventory led to the conclusion that the instrument may not tap cultural universals in the mother infant relationship. PMID- 11881715 TI - Inclusion of children in clinical research: lessons learned from mothers of diabetic children. AB - Nine mothers of diabetic children participated in a qualitative study about the factors that influenced them to consent to have their children involved in clinical research. They were asked to describe how they made decisions about involving their children in research, what motivated them to keep their children in research once they were enrolled, and how they evaluated the clinical studies their children had been in. Results suggest that mothers engage in a personal calculus before making a choice to consent. PMID- 11881716 TI - Social support and well-being in early adolescents: the role of mediating variables. AB - The purpose of this study was to examine the relationship between perceived social support and general well-being in early adolescents and to test two variables, hopefulness and self-esteem, that theoretically mediate this relationship. The final sample consisted of 142 adolescents, ages 12 to 14, who responded to instruments measuring perceived social support hopefulness, self esteem, and general well-being in classroom settings. Correlational analysis supported the five hypothesized relationships. A series of regression analyses described by Baron and Kenny indicated that hopefulness and self-esteem each were mediators of the relationship between social support and well-being. Implications for nursing practice are addressed. PMID- 11881717 TI - Development process and qualitative evaluation of a program to promote the mental health of family caregivers. AB - The purpose of this study was to develop and evaluate, through a participatory approach, an intervention program to promote the mental health of women caregivers in institutions. Focus groups were first organized to explore sources of stress for daughter and spousal caregivers. Workshops in which caregivers developed the content of a group program were then organized. The aims of the program were to increase empowerment and self-efficacy and to decrease stress and psychological distress of caregivers. Experimentation and qualitative evaluation of the program led to 10 weekly meetings covering the following topics: how to feel good with my relative, how to discuss with staff, how to appraise my experience differently, coping with my relative's losses, how to reorganize my life, and how to identify and ask for social support. This study provides a structured program to support family caregivers and a research method empowering families. PMID- 11881718 TI - Effects of crossing legs on blood pressure measurement. AB - The purpose of this study was to determine if blood pressure measurement is affected by the leg crossed at the knee as compared with feet flat on the floor in a well-senior population. Participants (N = 110) either had their blood pressure measured with feet flat first and then crossed or the reverse of this. Results indicate that blood pressure was significantly higher when legs were crossed versus uncrossed. Systolic pressure changed by 5.9 mmHg, from 127.32 to 133.24, whereas diastolic pressure changed by 2.97, from 72.54 to 75.52. There were no significant differences between those who had their blood pressure measured first with their legs crossed versus uncrossed or between those with and without hypertension. Instructing patients to keep feet flat on the floor during blood pressure measurement is an important nursing intervention that can contribute to the accurate measrement, interpretation, and treatment of a patient's health condition. PMID- 11881719 TI - HIV/AIDS knowledge and self-esteem among adolescents. AB - The incidence of HIV/AIDS is rapidly increasing among adolescents and young adults with some studies linking sexual risk taking and self-esteem. A convenience sample of 39 ethnically diverse adolescents, ages 14-18, participated in a pilot study designed to assess HIV/AIDS knowledge and to build self-esteem. Adolescents selected from two centers in California completed the Coopersmith Self-Esteem Inventory and the Student Health Questionnaire (SHQ) before beginning and after completing a program of six 2-hour educational sessions. These sessions focused on HIV/AIDS knowledge and building self-esteem. Knowledge of HIV/AIDS prevention and transmission increased by 2096 from pretest to posttest. Practitioners addressing the needs of adolescents should focus on in-depth information regarding HIV/AIDS, especially in the area of prevention strategies and cultural factors influencing levels of self-esteem. PMID- 11881720 TI - Diversity in a global society. PMID- 11881721 TI - Breast cancer care: providing practical and emotional support. PMID- 11881722 TI - General practitioners in North Wales: current experiences of palliative care. AB - Much work on the role of general practitioners within palliative care has focused on those working in densely populated inner city areas. This study was undertaken to evaluate the provision of palliative care and training needs of general practitioners in North Wales, a predominantly rural area. Questionnaires were sent to all 240 GPs and 94 responded (40%); those GPs responding were younger and mainly worked within partnerships. During the preceding year a median of four terminally ill patients had been cared for by each GP (range 0-21). General practitioners believed that palliative care was an important part of their role and applicable to patients with all end stage diseases. Use was made of community hospital beds as these were perceived as being more convenient for the patients and allowed the GPs to continue their involvement in patient care. Where services -e.g. palliative care consultants, day care and Marie Curie care--is available this was perceived as being very useful and of benefit. General practitioners believed they would benefit from further education and teaching on all aspects of palliative care, and this was supported by just over 50% of doctors knowing the correct breakthrough doses of morphine and many stating they would not prescribe more than a certain dose of opiates. Training in palliative care during vocational training was poor or non-existent and a preference was expressed for experiential teaching. PMID- 11881723 TI - Women's experience of breast conserving treatment for breast cancer. AB - The implementation of the National Breast Screening Programme in the UK, and subsequent increase in early detection of small cancers, has facilitated breast conserving treatment for more women with a breast cancer diagnosis than ever before. While a substantial body of literature has evolved regarding psychosocial morbidity and support needs of women at diagnosis and during treatment, there are specific gaps in the current knowledge base. The purpose of this study was to describe the experiences of women who had breast conserving treatment for early breast cancer, focusing on issues related to diagnosis, surgery, and radiotherapy. The study was designed within the context of clinical audit, with a view to informing service development. Seventy-six women who had undergone breast conserving surgery within the last 3-12 months, completed a self-report questionnaire. Findings indicated that although the majority of women expressed satisfaction with their treatment overall, a number of specific areas require attention from healthcare professionals. Further research is required to validate these findings and to explore: potential implications of different referral routes; information and support needs preceding definitive diagnosis; particular needs of those women with ductal carcinoma in situ (DCIS) vs. invasive disease; 'end of treatment' and ongoing information and support needs. PMID- 11881724 TI - Anxiety and depression in Iranian breast cancer patients before and after diagnosis. AB - A prospective study was conducted to measure anxiety and depression in Iranian breast cancer patients before and after diagnosis using the Hospital Anxiety and Depression Scale (HADS). The HADS was administered at two points in time: before diagnosis and 3 months after. In all, 168 breast cancer patients were interviewed. While 48% of patients had severe symptoms of anxiety at both baseline and follow-up, more than 60% of patients had no symptoms of depressive illness at pre-and post-diagnosis assessments. Comparing anxiety and depression before diagnosis and after 3 months, there were no significant differences between patients' scores on anxiety (P = 0.42) and depression (P = 0.98) subscales. The results showed that patients with advanced disease and a lower performance status were more anxious and experienced more depression. The study findings suggest that severe symptoms of anxiety are the most frequent symptoms in Iranian breast cancer patients. It seems that during the process of diagnosis and 3 months after psychological morbidity persists in patients who suffer from breast cancer. PMID- 11881725 TI - Treatment decision-making and its relation to the sense of coherence and the meaning of the disease in a group of patients with colorectal cancer. AB - The aims of the present study were to describe the preferred and the actual participating roles in treatment decision-making in relation to patients with newly diagnosed, colorectal cancer and to relate this result to the sociodemographic data, the Sense of Coherence Scale (SOC) and the patients' meaning of the disease. Eighty-six patients were studied. The following instruments were used: the Control Preferences Scale (CPS); the eight Lipowski categories of the meaning of the disease (LCMD); and the SOC. The results showed that 62% of the patients preferred a collaborative role and 28% a passive role in treatment decision-making. Agreement between the preferred and the actual participating roles was achieved by 44% of the patients. Seventy-one per cent of the patients showed an optimistic understanding of their disease. The mean SOC score was 150. There was no statistically significant difference between the CPS groups as regarded the sociodemographic data, the SOC and the LCMD. CONCLUSION: Sociodemographic data, the perceived meaning of the disease as well as the patients' sense of coherence were not related to the decision-making preferences in the investigated group of patients. Therefore, further investigations are needed to get an understanding of influencing factors of the decision-making preferences. PMID- 11881726 TI - Cancer fatigue. PMID- 11881727 TI - Morphological spectrum of cyclin D1-positive mantle cell lymphoma: study of 168 cases. AB - Immunostaining for cyclin D1 is essential for reliable diagnosis of mantle cell lymphoma (MCL). However, a small number of cyclin D1-positive lymphomas other than MCL have been encountered. Our goal was to investigate the morphological spectrum of MCL as a disease entity, based on cyclin D1 overexpression. We reviewed 181 biopsy specimens obtained from 168 cases of cyclin D1-positive MCL. Typical findings were the presence of nodular (53.9% of cases) or diffuse (46.1%) histological patterns, containing mantle zone patterns (16.8%), naked germinal centers (33.5%) and perivascular hyaline deposition (83.2%). Unusual findings of residual germinal centers with a mantle cuff (four cases) and follicular colonization (two cases) were seen. High magnification showed a monotonous proliferation of tumor cells with cytological diversity including small (3.0%), intermediate (43.1%), medium (34.1%), medium-large (13.2%) and large (6.6%) cells. Pleomorphic and blastic/blastoid variants were encountered in 9.6 and 7.2% of cases, respectively. Three cases had foci of cells of considerable size, with a moderately abundant pale cytoplasm resembling marginal zone B cells. Two cases showed an admixture of cells which appeared transformed and mimicked the histology of chronic lymphocytic leukemia/small lymphocytic leukemia. In one, neoplastic mantle zones were surrounded by sheets of mature plasma cells, resembling the plasma cell type of Castleman's disease. An admixture of areas characteristic of MCL and of other larger cells, indicating histological progression or a composite lymphoma, were observed in seven cases. In high-grade lesions of five cases, nuclear staining of cyclin D1 was rarely detected. In our experience, cyclin D1 expression was also found in nine lymphomas other than MCL (five plasma cell myelomas, three Hodgkin's disease and one anaplastic large cell lymphoma). The application of cyclin D1 staining prompted us to recognize the broad morphological spectrum of MCL. MCL can be diagnosed with the application of cyclin D1 immunostaining, if careful attention is given to architectural and cytological features. PMID- 11881729 TI - Immunohistochemical analysis of cyclooxygenase (COX)-2 expression in pancreatic endocrine tumors: association with tumor progression and proliferation. AB - An immunohistochemical study of cyclooxygenase (COX)-2 expression in pancreatic endocrine tumors (PET) was carried out, and the expression of COX-2 was compared with pathological features, the expression of several markers (hormones, vascular endothelial growth factor, single-stranded DNA, and the Ki-67 labeling index [LI]). Twenty PET, including 10 metastasizing cases (tumor size: 3-8 cm) and 10 non-metastasizing cases (tumor size: 0.3-8 cm) were studied. Tumors with a high level of COX-2 expression were placed in the H group, and the remaining tumors were placed in the L group. The H group was comprised of 13 tumors: all 10 of the metastasizing cases and three of the non-metastasizing cases. There were significant differences in tumor size between the two groups (H group 46.5 mm; L group 0.9 mm). There were significant differences in the presence of the following histological criteria for malignancy: pleomorphism (H group 13/13; L group 1/7), mitotic activity (H group 2.9; L group 0) and/or angioinvasion (H group 13/13; L group 1/7); and there were also significant differences in the number of cases that expressed ectopic hormones (gastrin, vasoactive intestinal peptide, serotonin and calcitonin; H group 12/13; L group 2/7) and in the Ki-67 LI (H group 8.3%; L group 0.4%). The distribution of COX-2-positive cells tended to be similar to the distribution of Ki-67-positive cells. Our data show that COX 2 is frequently upregulated in malignant PET and that there is a close relationship between COX-2 expression and tumor progression/proliferative activity. PMID- 11881728 TI - Increased expression of cyclooxygenase-2 in human salivary gland tumors. AB - We examined the immunohistochemical localization of cyclooxygenase (COX)-2 in human salivary gland tumors. Thirty salivary gland adenomas (SGA), 40 salivary gland carcinomas (SGC) and 15 normal salivary glands (NSG) were studied. NSG showed restricted COX-2 staining only in the epithelial cells of salivary ducts. In contrast, COX-2 protein was detected in 27 cases of SGA (90%), except for three myoepitheliomas, and in all cases of SGC (100%) at various intensities and in various fashions. Thirteen SGA (43%) and 36 SGC (90%) cases showed strong COX 2 staining predominantly in tumor cells containing ductal components, as did serous and mucous acinic components of acinic cell carcinomas, mucoepidermoid carcinomas and mucinous carcinomas. These findings may suggest that COX-2 in salivary gland tumors is expressed in tumor cells derived from pluripotential ductal epithelium that can histologically develop into either serous or mucinous acinar cells. PMID- 11881731 TI - Detection of numerical alterations of chromosome 1 in cytopathological specimens of breast tumors by chromogen in situ hybridization. AB - To investigate the effectiveness of chromogen in situ hybridization (CISH) in the diagnosis of breast tumors, numerical alterations of chromosome 1 were examined by CISH and fluorescence in situ hybridization (FISH) methods, and the presence of der(16)t(1;16) was also examined by FISH in imprinted cytology specimens from resected tissues of 14 carcinomas and five non-malignant lesions. The modal signal counts of chromosome 1 were compared between the specimens processed by CISH and FISH for each case. Aneusomies of the long arm of chromosome 1 were detected in 10 (71%) carcinomas as the major clones by both methods. In addition, one atypical papilloma demonstrated tetrasomy of 1q12 as a major clone by CISH, but such a clone was at first overlooked by FISH. Four other benign lesions showed disomic 1q12 signals as a major clone by both CISH and FISH. As additional information from FISH, eight cancers showed structural or numerical alterations of chromosome 16, and four showed der(16)t(1;16). In total, 10 carcinomas showed chromosome 16 alterations, and all of these overlapped with the carcinomas with 1q12 aneusomies. The CISH method provided almost the same results as the FISH method, and both methods were considered applicable in supportive diagnosis of cytological specimens of breast tumors. In addition, the CISH method was superior in the detection of numerical alterations in carcinoma cells by referring to the morphology of cells and in the detection of significant clones which might be missed under dark-field microscopy. PMID- 11881730 TI - Presence of natural killer-cell clones with variable proliferative capacity in chronic active Epstein-Barr virus infection. AB - Chronic active Epstein-Barr virus infection (CAEBV) is a syndrome that takes diverse clinical courses and is often associated with lymphoproliferative disorders of T/natural killer (NK)-cell lineage. We describe a patient with CAEBV associated with persistent pharyngeal ulcer, and with subsequent nasal T/NK-cell lymphoma in her neck lymph nodes and nasopharynx. Immunophenotyping of lymphoid cells showed that the lineage of Epstein-Barr virus (EBV)-positive cells in the patient was of NK-cell origin. By means of high-dose recombinant interleukin-2, we established an EBV-positive cell line of NK-cell lineage from her peripheral blood. Southern blot analysis for the number of terminal repeat sequences of EBV detected three NK-cell clones in the patient's lymph node. One of these clones was identical to the established cell line but was not observed in the pharyngeal ulcer, while the other two clones were present in the pharyngeal ulcer. These results suggest that the patient had expansion of the three NK-cell clones, one of which had proliferative capacity in vitro and was involved in the formation of the lymphoma. Moreover, the results suggest that the proliferative capacity of EBV-positive cells can be variable even in a single patient, and this variability may explain the clinical diversity in CAEBV. PMID- 11881732 TI - Leydig cell tumor of the testis: comparison of histopathological and immunohistochemical features of three azoospermic cases and one malignant case. AB - Leydig cell tumors of the testis are rare, mostly presenting as a testicular mass or as endocrinological symptoms. Here, three patients who were admitted for investigation of primary infertility and one patient presenting with a testicular mass are reported. The histological features were reviewed and an immunohistochemical study was done using a panel of antibodies against cytokeratin, vimentin, inhibin A, S-100, Ki-67, follicle-stimulating hormone, luteinizing hormone, prolactin, p53, bcl-2, and c-erbB2. The latter case (lost during follow up of metastatic disease) demonstrated massive tumor necrosis, extension through the tunica albuginea, and a high mitotic activity and MIB-1 score. Only this malignant case was bcl-2 positive. Of the two oncogenic markers studied, none of the cases were positive for c-erb2, while p53 was positive in more than 50% of cells in the malignant case and in one case of infertility with a large tumor, hemorrhage, focal necrosis and atypical cytological features. We recommend the evaluation of infertile men for Leydig cell tumors, and we believe that a panel of antibodies, including Ki-67, p53 and bcl-2, used for immunohistochemical analysis could be of diagnostic value in the identification of malignant and borderline cases of Leydig cell tumor. PMID- 11881733 TI - Mycobacterium tuberculosis infection within Warthin's tumor: report of two cases. AB - We report two patients with Warthin's tumor who were also infected with Mycobacterium tuberculosis. Case 1 was a 75-year-old woman with Warthin's tumor and multiple small epithelioid granulomas with caseous necrosis involving the submandibular gland. This patient died of tuberculous meningitis 4 months after biopsy. Case 2 was a 78-year-old man with a 10-year history of a parotid mass which had enlarged rapidly over 2 months. Surgical excision revealed Warthin's tumor and epithelioid granulomas involving the left parotid gland. DNA extracted from paraffin sections was amplified by nested polymerase chain reaction (PCR) with primer sets for the mycobacterial 65-KDa antigen gene. Restriction enzyme digestion of the PCR products could differentiate Mycobacterium tuberculosis from other mycobacteria in both cases. Although the histogenesis of lymphoid components of Warthin's tumor is controversial, the frequent prevalence of inflammation or necrosis and our present findings suggest these components have a similar behavior to regional lymph nodes. PMID- 11881734 TI - Autopsy case of lymphoplasmacytic lymphoma with a large submucosal tumor in the stomach. AB - An autopsy case of lymphoplasmacytic lymphoma with a large submucosal tumor in the stomach is presented. The patient was a 77-year-old woman with gastric lymphoma associated with Waldenstrom's macroglobulinemia of IgM-lambda type. Diagnosis was initially mucosa-associated lymphoid tissue (MALT) lymphoma of the stomach, because gastric biopsy specimens showed epitheliotropic proliferation (lymphoepithelial lesion) of the lymphoma cells. Postmortem examination revealed a large gastric lymphoma with metastatic foci in the esophagus, larynx, trachea, lungs, spleen and lymph nodes. The bone marrow was also involved. Lymphoma cells consisted of small lymphocytoid cells occasionally admixed with blast-like large cells and a large number of plasmacytoid or plasma cells. Centrocyte-like cells were not found. Lymphoepithelial lesions were not conspicuous in autopsy specimens. Immunohistochemically, lymphoma cells reacted with CD20, CD45, CD79a, anti-IgM, anti-lambda protein and anti-BCL-2, but not with CD5, CD10, CD23 or CD38. Based on these findings, the revised diagnosis of the present case was lymphoplasmacytic lymphoma, and it highlighted the differential diagnostic problem from marginal zone B-cell lymphoma of MALT type. PMID- 11881735 TI - Malignant lymphoma of bronchus-associated lymphoid tissue (BALT) coexistent with pulmonary tuberculosis. AB - A case in which malignant lymphoma occurred in association with a tuberculosis focus in a 70-year-old man is reported. Surrounding the epithelioid cell granulomas with caseous necrosis was a dense and diffuse monotonous infiltration of atypical lymphoid cells. Acid-fast bacilli were found in the granulomas and pulmonary tuberculosis was diagnosed. The infiltrating atypical lymphoid cells occasionally invaded the respiratory epithelium producing lymphoepithelial lesions. Immunohistochemically, the lymphoid cells were positive for CD20, and clonal rearrangement of the immunoglobulin heavy chain gene was demonstrated by polymerase chain reaction (PCR). We diagnosed the lesion as a pulmonary malignant lymphoma of bronchus-associated lymphoid tissue (BALT) occurring in the background of tuberculosis. This is the first reported case of pulmonary BALT lymphoma coexistent with pulmonary tuberculosis. PMID- 11881736 TI - Localized pleural malignant mesothelioma. AB - Pleural malignant mesothelioma (PMM) is a rare tumor and it is commonly seen in the form of multiple nodules or a diffuse tumor. A localized tumor mass in the pleura is extremely rare. Only seven cases have been reported. In this report, we present an additional case of localized PMM and describe the immunohistochemical and flow cytometric findings. A 61-year-old woman, without a history of smoking or asbestos exposure, presented with a severe pain in her right shoulder and arm. Chest radiography showed a solitary mass in the right upper lung field. Computed tomography showed a 5 cm right upper lung mass. Magnetic resonance imaging showed that the mass extended to the wall of the thorax. The patient underwent surgery for total removal of the tumor. Pathology revealed a localized malignant mesothelioma. Immunohistochemical analysis showed that the tumor was strongly and diffusely positive for cytokeratins with high and low molecular weight, and focally positive for vimentin and epithelial membrane antigen (EMA), but it was negative for carcinoembryonic antigen, Factor VIII, alpha-fetoprotein and Leu-M1. Flow cytometry showed an aneuploid DNA content in the tumor. The final diagnosis was localized malignant mesothelioma (epithelial type). The patient showed signs of local recurrence 5 months after surgery, and radiotherapy was given. PMID- 11881737 TI - Bizarre parosteal osteochondromatous proliferation (Nora's lesion) of the foot. AB - A 22-year-old man presented with a growing lump on the fifth metatarsal of the right foot. Radiographically, the lesion was a calcified mass stuck on to the bone. The T2-weighted magnetic resonance images showed heterogeneity in intensity. A tumor was suspected and an excisional biopsy was done. The lesion was composed of a cartilaginous cap and bone tissue. Histological examination revealed characteristic features of bizarre parosteal osteochondromatous proliferation (BPOP), such as hypercellularity, a blue tinctorial quality in the osteocartilaginous interfaces, and a scattering of binucleated or bizarre enlarged chondrocytes. Immunohistochemically, basic fibroblast growth factor was expressed in nearly all chondrocytes within the cartilaginous cap, while vascular endothelial growth factor was expressed only in enlarged chondrocytes near the osteocartilaginous interfaces. Reverse transcription-polymerase chain reaction detected chondromodulin-I transcripts in the tissue of the cartilaginous cap. These findings indicate that the processes occurring in BPOP are similar to those occurring in endochondral ossification in the growth plate, and they support the concept that BPOP is a reparative process. BPOP is a rare tumorous lesion of the bone and is occasionally confused with other benign or malignant conditions. Thus, it is important to consider the clinical, radiographical and the gross histological features of the lesion when making a diagnosis. PMID- 11881738 TI - Extra-abdominal desmoid tumor presenting as an intrathoracic tumor: case report and literature review. AB - A case of an extra-abdominal desmoid tumor presenting as an intrathoracic tumor (intrathoracic desmoid tumor) in a 46-year-old woman is reported. The tumor originated in the left chest wall and protruded into the left pleural cavity. Simple resection was carried out. The tumor, measuring 13 x 9 x 7 cm, was solid, gray-tan in color, and covered with parietal pleura. Histologically, the tumor was composed of a hypocellular arrangement of spindle-shaped cells with a fibromyxoid background. In some areas, keloid-like hyalinized collagen fibers proliferated, and a perivascular hypercellular area was seen. Immunohistochemical analysis showed that the cytoplasms of the tumor cells were strongly positive for vimentin, and some tumor cells were positive for alpha-smooth muscle actin, but all tumor cells were negative for CD34. These findings were consistent with the characteristics of an intrathoracic desmoid tumor. The differential diagnoses, in particular solitary fibrous tumor and tumors with a myofibroblastic nature, are discussed. PMID- 11881739 TI - Teratoid carcinosarcoma of the ovary with prominent neuroectodermal differentiation. AB - We present what we believe to be only the second report of ovarian teratoid carcinosarcoma. The patient, a 59-year-old woman, was admitted to hospital complaining of a pelvic mass and of abdominal fullness. Advanced ovarian cancer was diagnosed, and a tumorectomy was done. The tumor occupied the pelvis, and metastasis was found in the liver and spleen. The solid tumor was composed of chondrosarcoma, squamous cell carcinoma, adenocarcinoma and malignant neuroectodermal components, which contained ganglioneuroblastoma-like and medulloepithelioma-like areas. Immunohistochemically, the neuroectodermal cells were positive for both neural and epithelial markers. This ovarian tumor consisted of frankly malignant components, with prominent neuroectodermal elements mixed with epithelial and mesenchymal elements in an organoid fashion; a quite rare tumor. PMID- 11881740 TI - Superoxide dismutase and the death of motoneurons in ALS. AB - Amyotrophic lateral sclerosis (ALS) is a lethal disease that is characterized by the relentless death of motoneurons. Mutations to Cu-Zn superoxide dismutase (SOD), though occurring in just 2-3% of individuals with ALS, remain the only proven cause of the disease. These mutations structurally weaken SOD, which indirectly decreases its affinity for Zn. Zn-deficient SOD induces apoptosis in motoneurons through a mechanism involving peroxynitrite. Importantly, Zn deficient wild-type SOD is just as toxic as Zn-deficient ALS mutant SOD, suggesting that the loss of Zn from wild-type SOD could be involved in the other 98% of cases of ALS. Zn-deficient SOD could therefore be an important therapeutic target in all forms of ALS. PMID- 11881741 TI - Gamma-secretase: never more enigmatic. AB - It is widely believed that the pathogenesis of Alzheimer's disease (AD) is intimately, if not causatively, associated with the deposition of approximately 4 kDa beta-amyloid (A beta) peptides in the cerebral cortex and hippocampus of affected individuals. A beta peptides are liberated from transmembrane proteins, termed beta-amyloid precursor proteins (APP), by the concerted action of beta- and gamma-secretase(s). Whereas the identity of beta-secretase is no longer in question, the identity of gamma-secretase, which is responsible for the intramembranous processing of APP, has never been more enigmatic. Considerable evidence has accrued to impugn the presenilins (PS) as the executioners of intramembranous processing of APP. Here, we summarize these observations and review recent evidence that argues against the prevailing hypothesis that PS function as gamma-secretases. PMID- 11881742 TI - Molecular mechanisms of brain aging and neurodegenerative disorders: lessons from dietary restriction. AB - The application of modern molecular and cell biology technologies to studies of the neurobiology of aging provides a window on the molecular substrates of successful brain aging and neurodegenerative disorders. Aging is associated with increased oxidative stress, disturbances in energy metabolism and inflammation like processes. Dietary restriction (DR) can extend lifespan and might increase the resistance of the nervous system to age-related neurodegenerative disorders. The neuroprotective effect of DR involves a preconditioning response in which the production of neurotrophic factors and protein chaperones is increased resulting in protection against oxyradical production, stabilization of cellular calcium homeostasis, and inhibition of apoptosis. DR might also enhance neurogenesis, synaptic plasticity and self-repair mechanisms. PMID- 11881743 TI - Huntington's disease: new hope for therapeutics. AB - Huntington's disease (HD) is one of eight progressive neurodegenerative disorders in which the underlying mutation is a CAG expansion encoding a polyglutamine tract. There are currently no cures or even effective therapies for HD. Effective strategies have remained elusive because little is known about either the mechanisms of expansion or the mechanism of polyglutamine-mediated neuronal death. However, recent advances in understanding the basic mechanisms of expansion and toxicity have renewed hope that a therapeutic strategy might someday be possible. Strategies effective in the treatment of HD are likely to be relevant in the treatment of a range of neurological and neurodegenerative disorders. PMID- 11881744 TI - Humoral autoimmunity as a mediator of CNS repair. AB - Autoimmune responses directed against the central nervous system (CNS) have generally been considered pathogenic in nature. Although there are several well understood conditions in which this is the case, there is also a growing body of experimental evidence to show that both the cellular and humoral immune responses can promote tissue repair following CNS injury and disease. Our laboratory has used a mouse model of chronic demyelinating disease to characterize a class of polyreactive IgM autoantibodies that react with oligodendrocyte surface antigens and promote myelin repair. By screening a large number of human monoclonal antibodies, we have found that IgM antibodies that react with CNS tissue are relatively common. Autoreactive IgM antibodies might constitute an endogenous system for tissue repair, and therefore these antibodies could be of value as therapeutic reagents. PMID- 11881745 TI - Cholesterol, A beta and Alzheimer's disease. AB - Statins have been used for many years for the treatment of hypercholesterolemia. They lower low-density lipoprotein (LDL) cholesterol, increase high-density lipoprotein (HDL) levels and are considered to be very safe. Recently, a set of potential new applications was identified for statins. In the future, these drugs could be used to treat Alzheimer's disease (AD). Past studies have suggested a link between AD and lipids and a series of reports has recently been published that significantly tightens this link and also provides some explanations at the cellular level. This review focuses on these recent developments and perspectives that appear to link cholesterol, beta-amyloid and AD. PMID- 11881746 TI - Engineered modeling and the secrets of Parkinson's disease. AB - The development of new methods for manipulating the animal genome by transgenic and gene-targeting technologies provides a unique means of studying the most intimate aspects of countless human diseases, including Parkinson's disease (PD). In this review, the contribution of such engineered models to our current understanding of the pathophysiology, etiology and pathogenesis of PD will be discussed. PMID- 11881747 TI - Antisense as a neuroscience tool and therapeutic agent. AB - Gene expression in the mammalian brain is highly complex and requires an immensely powerful functional genomics tool to unravel it. Antisense has the potential to meet this requirement, but has always been plagued by biological and technological hurdles that have made the technology unreliable. With recent progress in developing potent, low-toxicity nucleic acid chemistries and novel drug delivery methods to cross the blood-brain barrier, the use of antisense is gathering momentum. PMID- 11881748 TI - The role of the ubiquitin-proteasomal pathway in Parkinson's disease and other neurodegenerative disorders. AB - A unifying feature of neurodegenerative diseases is the abnormal accumulation and processing of mutant or damaged intra- and extracellular proteins; this leads to selective neuronal vulnerability and dysfunction. The ubiquitin-proteasomal pathway (UPP) is poised to play a central role in the processing of damaged and toxic proteins by ubiquitin-dependent proteolysis. The UPP can be overwhelmed in several neurodegenerative diseases. This results in the accumulation of toxic proteins and the formation of inclusions, and ultimately to neuronal dysfunction and cell death. Further analysis of the cellular and molecular mechanisms by which the UPP influences the detoxification of damaged and toxic proteins in neurodegenerative diseases could provide novel concepts and targets for the treatment and understanding of the pathogenesis of these devastating disorders. PMID- 11881749 TI - Comparison of comfort and local complications after cardiac catheterization. AB - The purpose of this study was to compare the effects of 4 hours of bed rest versus 6 hours of bed rest on patients' safety, comfort, and satisfaction levels. Using a quasi-experimental design, the authors studied 118 left-heart catheterization patients who were randomly assigned to 4 hours or 6 hours of bed rest. Among the study participants, only 1 in the 6-hour group had significant bleeding. There were no complications in the 4-hour group. There were no statistically significant differences between the two groups on any of the other study variables. Given the lack of significant complications for the 4-hour group and similar comfort levels for both study groups, these findings suggest the feasibility of reducing the standard period of postcatheterization bed rest from 6 hours to 4 hours, thereby possibly lowering the cost of the outpatient procedure. PMID- 11881750 TI - Determining stroke rehabilitation inpatients' level of nursing care. AB - The purpose of this study was to evaluate the feasibility of using the Functional Independence Measure (FIM) to predict staffing needs of stroke patients in an acute inpatient rehabilitation program. The Patient Care Index (PCI) was concurrently administered with the FIM on all stroke admissions to a stroke rehabilitation unit over a 3-month period. One hundred fourteen patients 18 years of age or older admitted to the unit with a medical diagnosis of stroke were included in the sample. Total FIM score had a strong inverse relationship to the level of care indicated by the PCI at Days 1, 5, 7, 10, 15, and 20 of rehabilitation (r(s) = -. 76 to -.87). Total FIM score and the need for staff supervision for safety were the two factors predictive of the level of nursing care. The FIM has potential to guide nurse-staffing decisions. PMID- 11881751 TI - Caregivers in crisis: moving a relative with Alzheimer's to assisted living. AB - The experience of moving a relative with Alzheimer's disease (AD) to an assisted living facility (ALF) was examined in this secondary data analysts of 20 interviews with family caregivers. Crisis theory was used as guiding framework for the analysis. Findings indicate that all elements of a crisis were evident among caregivers in the process of moving their relative to an ALF. Dangerous behaviors resulting from the relative's cognitive decline led to caregiver fear and anxiety, which served as crisis precipitants. Perceived lack of family support in conjunction with physical and psychological exhaustion were crisis mediators. These factors, along with the caregiver's failed attempts to keep their relative at home, caused the crisis process to move toward the outcome of ALF placement. A better understanding of the crises that surround placement issues will assist nurses in planning interventions for the growing number of families faced with this issue. PMID- 11881752 TI - Psychosocial impact of heart transplantation on spouses. AB - The purpose of this study was to evaluate the psychosocial impact of heart transplantation on spouses and compare the adjustment of spouses and their partners. Data were collected from 51 couples prior to transplant and 12 months posttransplant. The Psychosocial Adjustment to Illness Scale (PAIS) was used to measure adjustment in seven domains. Prior to the transplant, spouses experienced profound psychosocial distress related to their partner's illness. From pretransplant to posttransplant, PAIS total score decreased (lower scores indicate better adjustment) for spouses (M = 42, SD = 16, vs. M = 26, SD = 13). Spouses showed improvement in all domains. Posttransplant, patients' and spouses' PAIS total scores (overall adjustment) were not significantly different. Spouses reported more psychological distress than patients; patients reported more problems than spouses in vocational and domestic function. Study findings highlight the importance of sensitivity in the clinical setting to the distinct psychosocial needs of spouses. PMID- 11881753 TI - Referring abused women: does police assistance decrease abuse? AB - To evaluate if abused women's use of the police reduced further violence experienced, 90 abused women were interviewed at a large urban police department and followed for 6 months. Frequency and severity of violence as well as danger of homicide were measured before and at 3 and 6 months after women sought police help. Women seeking police help had significantly reduced threats of abuse (F = 124.62, df = 2,81; p < .0005), actual abuse (F = 90.11, df = 2,81; p < .0005), and danger of being killed (F = 188.69, df = 2,81; p < .0005) by her intimate partner. Bonferroni post hoc tests showed that significantly fewer threats of abuse, physical abusive acts, and risk indicators for homicide had occurred from prefiling to 3 months (p <.0005) and prefiling to 6 months (p <.0005). The data indicate that referring abused women to the police is an appropriate intervention. PMID- 11881754 TI - Association between autonomic and motoric systems in the preterm infant. AB - In the clinical setting, fetal and infant movement is used as an indicator of central nervous system and neurobehavioral developmental status. Current models of neurobehavioral development include the synactive theory of neonatal behavioral organization, which defines and describes the interaction between five subsystems. Results of testing synchronous interaction between two of those systems--the autonomic and motoric subsystems in preterm infants--are reported here. PMID- 11881755 TI - Economic pressure in African American families: a replication and extension of the family stress model. AB - This study of 422 two-caregiver African American families, each with a 10-11-year old focal child (54% girls), evaluated the applicability of the family stress model of economic hardship for understanding economic influences on child development in this population. The findings generally replicated earlier research with European American families. The results showed that economic hardship positively relates to economic pressure in families. Economic pressure was related to the emotional distress of caregivers, which in turn was associated with problems in the caregiver relationship. These problems were related to disrupted parenting practices, which predicted lower positive child adjustment and higher internalizing and externalizing symptoms. The results provide significant support for the family stress model of economic hardship and its generalizability to diverse populations. PMID- 11881756 TI - Skin-to-Skin contact (Kangaroo care) promotes self-regulation in premature infants: sleep-wake cyclicity, arousal modulation, and sustained exploration. AB - The effect of mother-infant skin-to-skin contact (kangaroo care, or KC) on self regulatory processes of premature infants was studied. Seventy-three infants who received KC were compared with 73 infants matched for birth weight, gestational age, medical risk, and family demographics. State organization was measured in 10 s epochs over 4 hr before KC and again at term. No differences between KC infants and controls were found before KC. At term, KC infants showed more mature state distribution and more organized sleep-wake cyclicity. At 3 months, KC infants had higher thresholds to negative emotionality and more efficient arousal modulation while attending to increasingly complex stimuli. At 6 months, longer duration of and shorter latencies to mother-infant shared attention and infant sustained exploration in a toy session were found for KC infants. The results underscore the importance of maternal body contact for infants' physiological, emotional, and cognitive regulatory capacities. PMID- 11881757 TI - Development of quantitative reasoning and gender biases. AB - One hundred twenty-seven 7-, 9-, and 11-year-old children were presented large or small samples of own-gender enhancing or other-gender enhancing observations. Children read arguments based on the observations, rated argument intelligence, judged the number of other children to whom the observations could be generalized, and provided verbal justifications for their judgments. Own-gender reasoning biases declined with age; these declines were, however, partially accounted for by declines in the strength of self-reported gender affiliations. Reasoning biases--demonstrated by problem-to-problem shifts in reasoning quality were constrained by sample size, indicating a modest degree of rationality even among 7-year-olds. Specifically, biases co-existed with reasonably limited generalizations from small samples of own-gender evidence and with reasonably extensive generalizations from large samples of other-gender evidence. Children were thus able to satisfy motivations for own-gender favoritism and reason in accord with the law of large numbers. Several explanations of the findings-based on changes in the salience of gender, multiple classification skills, and the ability to reason independently from beliefs-are offered. PMID- 11881758 TI - Anger regulation in disadvantaged preschool boys: strategies, antecedents, and the development of self-control. AB - Emotion regulation strategies observed during an age 3 1/2 frustration task were examined in relation to (a) angry affect during the frustration task, (b) child and maternal characteristics at age 1 1/2, and (c) indices of self-control at age 6 in a sample of low-income boys (Ns varied between 189 and 310, depending on the assessment). Shifting attention away from sources of frustration and seeking information about situational constraints were associated with decreased anger. Secure attachment and positive maternal control correlated positively with effective regulatory strategy use. Individual differences in strategy use predicted self-control at school entry, but in specific rather than general ways: Reliance on attention-shifting strategies corresponded with low externalizing problems and high cooperation; reliance on information gathering corresponded with high assertiveness. PMID- 11881759 TI - The effects of sex steroids on spatial performance: a review and an experimental clinical investigation. AB - Males typically surpass females in spatial performance, an outcome that may be linked to testosterone and estrogen. The authors (a) review physiological mechanisms, developmental periods, and past empirical work relevant to sex steroids' effects on human spatial performance and (b) report an experimental study of the role of actively circulating sex steroids in adolescents being treated for delayed puberty (N = 55; mean age = 13.70 years). Sex steroids (simulating early, middle, and late puberty) and placebos were given alternately over 21 months and spatial tests were given every 3 months. Spatial performance showed traditional sex differences but did not vary with levels of actively circulating sex steroids. PMID- 11881760 TI - "Remember where you last saw that card": children's production of external symbols as a memory aid. AB - Four experiments examined the age at which children start to use external symbols to aid their memory and how external symbol use affects both their memory performance and information allocation strategies. In Experiment 1, children in Grades 1, 3, 5, and 7 played a memory card game (Concentration) twice, once with the opportunity to make notes to aid performance and once without the opportunity. Grades I and 3 students tended to produce nonmnemonic notations, whereas Grades 5 and 7 students were more likely to produce functional, adultlike notations that aided performance in the task. In Experiments 2a and 2b, unexpected removal of children's notations led to a decrease in performance. suggesting that the spontaneously produced notations were being used as an external store rather than as an aid to encoding information. Experiment 3 examined whether all information was placed in external storage or if some types of information remained in memory. Grade 7 students who had their notations unexpectedly taken away were able to recognize the identity of the cards they had previously seen but had more difficulty remembering their locations. They appeared to place the location information mainly in external storage while retaining the identity information in memory. These results suggest that in mid childhood, children begin to distribute information actively between internal and external memory storage. PMID- 11881761 TI - Children's coping strategies: moderators of the effects of peer victimization? AB - Coping strategies were examined as potential moderators of the effects of peer victimization on children's adjustment. Self-report data on victimization experiences, coping strategies, and loneliness were collected on ethnically diverse 9-10-year-old children (177 girls, 179 boys). Teacher ratings of children's anxious-depressed and social problems and peer nominations of social preference were also obtained. Findings revealed that strategies such as problem solving that were beneficial for nonvictimized children exacerbated difficulties for victimized children. The effects of specific forms of coping were dependent on gender: social support seeking buffered victimized girls from social problems but was associated with lower peer preference for victimized boys. Data also revealed the need to examine the effects of coping on multiple adjustment outcomes. PMID- 11881762 TI - Age differences in the relationship between visual movement imagery and performance on kinesthetic acuity tests. AB - The Vividness of Movement Imagery Questionnaire was administered to 280 fourteen year-olds and 186 ten-year-olds, who were then tested on measures of kinesthetic acuity. The relationship between visual movement imagery and kinesthetic acuity scores was significant in the older group: Those with high levels of visual movement imagery performed significantly better on measures of kinesthetic acuity than did those with low imagery. No such effect was found in the younger group. The results indicate that for adolescents, the confounding effect of visual imagery affects the researcher's ability to interpret kinesthetic acuity scores. The relationship between imagery and kinesthesis appears to develop over the period between 10 and 14 years, although such an interpretation may be premature because the measurement of visual movement imagery in the younger age group is problematic. PMID- 11881763 TI - Developmental changes in mother-infant face-to-face communication: birth to 3 months. AB - This study documented the growth of the earliest form of face-to-face communication in 16 mother-infant dyads, videotaped weekly during a naturalistic face-to-face interaction, between 1 and 14 weeks, in 2 conditions: with the infant in the mother's arms and with the infant semi-reclined on a sofa. Results showed a curvilinear development of early face-to-face communication, with a significant increase occurring between Week 4 and Week 9 depending on the dyad. After 2 months, trajectories diverged into 2 groups: I whose duration of face-to face communication continued to increase and I whose duration peaked and then began to decrease. After the 1st month, the duration of face-to-face communication was significantly longer when the infant was on the sofa rather than in the mother's arms. In the latter condition, during the 3rd month, girls spent a significantly longer time than boys in face-to-face communication. These findings suggest that context (infant being held vs. not being held) interacts with the infant's age and sex in affecting mother-infant communication. PMID- 11881764 TI - Sex and contextual effects on children's use of interference competition. AB - Prior research with children and nonhuman animals suggests that females engage in interference competition, in which I individual reduces another's chances of gaining access to a resource, only when resources are scarce, whereas males use it more widely. This study was designed to compare males' and females' use of interference competition in games in which resources were scarce or plentiful. Forty groups of 4 same-sex children from kindergarten or Grade 4 played the 2 games on 2 days. Grade 4 girls used less interference competition when resources were plentiful than when they were scarce. Results are useful for generating a contextually based model of the development of sex differences in competitive behavior. PMID- 11881765 TI - Strategic attention deployment for delay of gratification in working and waiting situations. AB - Two studies examined whether the detrimental effects of attention to rewards on delay of gratification in waiting situations holds-or reverses-in working situations. In Study 1, preschoolers waited or worked for desired delayed rewards. Delay times increased when children worked in the presence of rewards but, as predicted, this increase was due to the distraction provided by the work itself. not because attention to rewards motivated children to sustain work. Analysis of spontaneous attention deployment showed that attending to rewards reduces delay time regardless of the working or waiting nature of the task. Fixing attention on rewards was a particularly detrimental strategy regardless of the type of task. Study 2 showed that when the work is not engaging, however, attention to rewards can motivate instrumental work and facilitate delay of gratification as long as attention deployment does not become fixed on the rewards. PMID- 11881766 TI - Optimal biopsy techniques in the diagnosis of primary ciliary dyskinesia. AB - OBJECTIVE: Adequate biopsy specimens that clearly demonstrate cilia, and therefore enable the determination of the presence or absence of primary cilia dyskinesia, may be difficult to obtain. This study is an attempt to identify the optimal sampling technique to best examine respiratory tract cilia. DESIGN: A prospective comparison of the four sampling techniques was carried out: nasal brushing, nasal biopsy, bronchial brushing, and tracheal biopsy. SETTING: Tertiary care pediatric hospital: Children's Hospital of Eastern Ontario. METHODS: Ten consecutive patients booked for bronchoscopy and tracheal biopsy underwent all four procedures. Specimens were examined under light microscopy for an assessment of quality. RESULTS: The nasal brushing and tracheal biopsy specimens provide superior quality (p = .22); however, nasal brushing is more cost efficient. Nasal biopsy samples frequently are metaplastic and therefore are inferior to nasal brushing samples (p = .02). CONCLUSION: With equal efficiency demonstrated, the reduction in potential morbidity and health care costs suggests nasal brushings to be the optimal initial investigation for primary ciliary dyskinesia. PMID- 11881767 TI - Indications for interventional arteriography in post-tonsillectomy hemorrhage. AB - OBJECTIVE: To determine the incidence of post-tonsillectomy hemorrhage (PTH) and required measures of treatment. DESIGN: Retrospective chart review. SETTING: Department of Otolaryngology-Plastic Head and Neck Surgery, St. Anna Hospital, Duisburg, Germany. METHODS: The charts of 5,539 patients at risk for PTH (post tonsillectomy) and seen in our department between 1988 and 1998 were reviewed including 65 patients primarily operated on elsewhere. MAIN OUTCOME MEASURES: Success and failure of surgical methods. RESULTS: Surgical treatment of PTH was necessary in 145 of our patients (2.65%). Owing to recurrent bleeding in 7 patients, 153 surgical procedures to stop bleeding became necessary. There was one case with lethal outcome (0.018%). Massive bleeding indicated ligature of the external carotid artery (ECA) in 5 patients (0.09%). Despite this procedure, hemorrhage recurred in another 3 patients, all of whom had undergone tonsillectomy elsewhere. In 2 of these cases, arteriography revealed arterial blood supply originating from branches of the internal carotid artery (ICA) or the carotid bulb. CONCLUSION: When ligation of the ECA is considered, a complete dissection of the ICA, ECA, and carotid bulb should be performed to exclude any vascular abnormality. In cases with unclear blood supply of the tonsillar lodge or continuous bleeding despite ligation of the ECA, immediate arteriography of the carotid artery should be indicated. If possible, simultaneous selective embolization of the bleeding vessel should be performed to replace a mutilating approach associated wit exposure and ligation of the ECA. PMID- 11881769 TI - Nasomaxillary frame translocation in medial maxillectomy. AB - Medial maxillectomy is the standard operation for en bloc resection of the lateral nasal wall with portions of the maxillary and ethmoid sinuses. Although most reports comment on good to excellent cosmetic results, nasal collapse is a possible complication of the procedure and is best explained by the loss of lateral nasal wall support. To overcome this problem, we describe a technique with temporary mobilization of a bone frame around the ipsilateral pyriform aperture, including one nasal bone, and fixing it to its original position at the end of the operation. PMID- 11881768 TI - Hearing outcome following laser-assisted stapes surgery. AB - This article reports the postoperative results of 76 operations addressing otosclerosis. All procedures were performed by the same surgeon and took place within a 4-year period. We compared the postoperative audiometric results of 54 laser-assisted procedures with those of 22 nonlaser operations. Our findings revealed a statistically significant improvement in the air-bone gap closure of the laser-operated patients: the air-bone gap was narrowed to within 10 dB in 72% of laser-operated cases versus 54% in nonlaser cases. Furthermore, laser use does not induce high-frequency sensorineural hearing loss. We believe that a laser entails less risk of inner ear microtrauma and improves the accuracy and reliability of stapedotomy. PMID- 11881771 TI - Endoscopic removal of localized angiofibroma. PMID- 11881770 TI - Solitary central nervous system metastasis from acinic cell carcinoma of the parotid gland. PMID- 11881772 TI - Nasal squamous cell carcinoma in a child. PMID- 11881773 TI - Chronic otitis media masking coexistent acoustic neuroma. PMID- 11881774 TI - Visual vestibular mismatch in patients treated with intratympanic gentamicin for Meniere's disease. AB - OBJECTIVE: To support the hypothesis that the symptom complex known as visual vestibular mismatch (VVM) can be induced by peripheral vestibular disease. DESIGN: Retrospective chart review; prospective questionnaire. SETTING: A tertiary/quaternary care hospital clinic. METHODS: The charts of 28 patients who were treated for Meniere's disease were studied. Their responses to a pretreatment VVM questionnaire were scored and compared with their answers in a telephone follow-up to the same questions posttreatment. These questionnaires were compared with those filled in by 100 control patients without ear disease. MAIN OUTCOME MEASURES: Patients' responses to a VVM-specific questionnaire. RESULTS: Seventeen of 28 patients had VVM. Gentamicin therapy increased the number of positive answers. There was no correlation between the development of VVM complaints, caloric scores, and posturography performance. No control patients had symptoms of VVM. CONCLUSION: We conclude that the development of VVM indicates the onset or worsening of vestibular disease as it can be induced or exacerbated by gentamicin therapy. As there is no correlation between VVM and caloric scores, we suggest that otolithic damage may be responsible for this symptom set developing. PMID- 11881775 TI - Ossification of the auricles: a forgotten sign in adrenal insufficiency. PMID- 11881776 TI - Revisiting Mycobacterium bovis. PMID- 11881777 TI - Primary nasal tuberculosis. PMID- 11881778 TI - Inferior concha bullosa. PMID- 11881779 TI - Hyposmia following laryngectomy: experimental model. AB - Hyposmia following laryngectomy is a well-known clinical observation, yet the causes have been controversial for many years. In an attempt to resolve this issue, an animal model was constructed. Eighteen dogs were divided into three equal groups: control dogs, dogs that underwent tracheostomy, and dogs that underwent tracheostomy and denervation of the larynx, simulating total laryngectomy. Four to 6 months following these operations, biopsies from olfactory mucosa were taken. The results showed marked changes in the olfactory mucosa of the two test groups: cystic degeneration of secretory glands in the olfactory mucosa of the first group and involution of the olfactory mucosa, substituted by dense connective tissue and "ballooning" of olfactory nerve fibres in the second group. These findings suggest that the changes in olfactory mucosa are not only caused by a loss of nasal airway but also the existence of a neurologic connecting network between the vagus nerve and the olfactory cortex. PMID- 11881780 TI - Word recognition deficits in German: more evidence from a representative sample. AB - In a representative sample of German speaking dyslexic children, earlier findings on dyslexia in the highly consistent orthography of German were confirmed. In a sample of 78 dyslexic 3rd graders selected on the basis of their poor word recognition skills, reading accuracy for both words and non-words was deficient but high in absolute terms. This indicates that the highly consistent grapheme phoneme correspondences of German orthography in combination with the straightforward phonics teaching approach, which is usually applied in Austrian primary schools, allows even dyslexic children to acquire the process of phonological decoding. The central reading problem was extremely slow speed. Poor performance on a reading comprehension test was at least partly due to slow reading speed as well. Dyslexic children's spelling development was also severely delayed. The number of phonologically incorrect spellings was low; however, dyslexic children quite often produced spellings that are orthographically incorrect, indicating that they have not yet been able to develop an extensive and easily accessible orthographic lexicon. The most prominent cognitive deficit was reduced rapid naming speed closely followed by deficits in phonological awareness. Deficits in phonological memory were also evident (but mild) showing that despite marked differences in findings on reading and spelling skills of English dyslexic children, the underlying causes are very similar. The present sample of dyslexic children showed deficits in visual processing speed in addition to their linguistic deficits. However, this deficit in visual processing speed did not seem causally related to the children's reading and spelling deficits. PMID- 11881781 TI - Outstanding questions about phonological processing in dyslexia. AB - It is widely accepted that developmental dyslexia results from some sort of phonological deficit. Yet, it can be argued that phonological representations and their processing have been insufficiently tested in dyslexia research. Firstly, claims about how tasks tap into certain kinds of representations or processes are best appreciated in the light of an explicit information-processing model. Here, a cognitive model of lexical access is described, incorporating speech perception, reading and object recognition. The model emphasizes that phonological forms of lexical items are distinct from non-lexical phonological representations. Secondly, phonology, as a linguistic discipline, teaches us that there is much more to it than phonemic categorization and awareness. The phonological level of representation also embodies phonotactic regularities, patterns of phoneme assimilation and alternation, as well as supra-segmental knowledge pertaining to syllable structure, stress, intonation and rhythm. All these aspects are in part language-dependent, and therefore must be learnt by children in order to become proficient native speakers and listeners. If phonological representations were affected in dyslexia, dyslexic children would presumably have difficulties acquiring these aspects of their language. This prediction is as yet untested. A possible research agenda is outlined, aiming to provide a more comprehensive assessment of the phonological theory of dyslexia. PMID- 11881782 TI - Differential effects of orthographic transparency on dyslexia: word reading difficulty for common English words. AB - Orthographic transparency is increasingly being recognised as an important factor in determining the manifestation of dyslexic tendencies in individuals. Recent evidence has shown that normal English-speaking children have reading deficits in the range associated with same age dyslexic German-speaking children for less frequently used words, and English orthography has been identified as a contributing factor. Spencer (Reading (1999) 33(2), 72-77; Journal of Research in Reading (1999) 22(3), 283-292) has proposed a predictive model for English children's reading and spelling deficits, based on orthographic features. Two main factors have been identified: consistency of sound representation and inclusion of redundant letters in English words. Using new data for reading, a unified model will be discussed which incorporates measures of depth of orthographic transparency and relates these to recent positron emission tomography (PET) research. Implications for the relative frequency and severity of dyslexia in both deep and shallow orthographies is considered in the light of such studies. PMID- 11881783 TI - Phonological spelling errors among dyslexic children learning a transparent orthography: the case of Czech. AB - Substantial evidence from studies of English-speaking dyslexic children's spelling suggests that these individuals have a persistent impairment in representing the phonological structure and content of words in writing. In contrast, several studies of German dyslexic children (Landerl & Wimmer, 2000) suggest that, among learners of transparent orthographies, the above impairment is transient and resolves by the end of grade 2; instead dyslexic spelling is characterised by a persistent impairment in learning inconsistent spelling patterns. In order to determine whether phonological spelling difficulties are transient among dyslexic spellers of other transparent orthographies, we analysed phonological accuracy in the spellings of 43 dyslexic children, 43 age-matched control and 43 spelling-matched control children attending primary schools in Prague. Czech dyslexic children presented an interesting test case because Czech has a transparent orthography. Czech dyslexics as old as 11 years (grade 5) continued to produce high rates of phonologically inaccurate spellings relative to their age peers; thus their difficulties with phonological representation in spelling had not resolved. This pattern is consistent with findings from studies of English-speaking dyslexics, and it suggests that orthographic depth may impact less dramatically on the manifestation of dyslexia than has been proposed previously. PMID- 11881784 TI - 'Reflections on StudyScan'. PMID- 11881785 TI - Patterns of abdominal relapse and role of sonography in Wilms tumor. AB - This study characterizes the patterns of abdominal recurrence of Wilms tumor and describes the role of sonography in its detection. Twelve patients who had initial tumor recurrence in the abdomen were evaluated. Five patients had recurrence in the kidney; all had nephrogenic rests detected by computed tomography (CT) or magnetic resonance (MR) imaging but not by sonography. The remaining 7 patients had recurrence in the peritoneum (4), the nephrectomy site (2), or the regional lymph nodes (1); tumor spillage had occurred in five of these patients. Four recurrences were detected during therapy, and eight within 3 years after completion of therapy. Seven of the 12 recurrences were first detected by sonography. All 11 sonograms obtained at the time of relapse showed tumor recurrence. Nine patients died a median of 10 months after relapse. The results suggest that regular sonographic surveillance for 3 years after therapy is likely to reveal most abdominal recurrences. Supplementation with CT or MR imaging is indicated for detection of nephrogenic rests. PMID- 11881786 TI - P53 gene mutations in pleuropulmonary blastomas. AB - Pleuropulmonary blastoma (PPB) is rare childhood tumor oniginating from either lung or pleura. Although several cytogenetic changes, such as tisomy 2, trisomy 8, and loss of 17p material, have been reported, evidence of gene mutations is still lacking. Pathologically, PPB shares similarities with rhabdomyosarcoma in which p53 mutations arefrequently detected. Possible implication of p53 mutations in PPB was investigated. PPBs of 3 patients were analyzed for occurrence of p53 mutations by using polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) method, and the nature of mutations was confirmed by direct sequencing. Two PPBs were confirmed to harbor p53 mutations. One was a Val to Leu substitution at codon 173, and another was a ArgArg to TrpCys substitution at codons 282 and 283. In each tumor, only the mutated allele was detected, suggesting inactivation of p53. Both patients with mutations had fatal outcome, while the remaining patient in whom no mutation was detected is disease free for 3 years after completion of treatment. The results raise the possibility that p53 inactivation can occur as a nonrandom genetic change involving the pathogenesis and outcome of PPB. Further studies in a larger series are necessary to clarify these matters. PMID- 11881787 TI - Spindle cell lesions of the head and neck mimicking rhabdomyosarcoma in children. AB - Malignancies of the head and neck are uncommon among children. The most common solid tumors to occur in this region are rhabdomyosarcoma, Hodgkin disease, and non-Hodgkin lymphoma. Two children are described who presented with signs and symptoms consistent with rhabdomyosarcoma, but who were found to have benign spindle cell lesions. These rare lesions should be part of the differential of children with a head and neck mass. PMID- 11881788 TI - Nephrotoxicity due to intermediate-dose methotrexate without rescue in an obese adolescent with acute lymphoblastic leukemia. AB - Toxicity from intermediate-dose methotrexate (MTX) is unusual. A severely obese adolescent with acute lymphoblastic leukemia experienced significant, delayed nephrotoxicity from intermediate-dose MTX. Altered MTX disposition may occur as a consequence of other ingestions or conditions such as obesity. The use of radioisotope renography established the diagnosis and followed the resolution of the patient's MTX-induced nephrotoxicity. PMID- 11881789 TI - Pneumocystis carinii pneumonia in a girl with a midbrain glioma. AB - A 7-year-old girl with a midbrain glioma contracted Pneumocystis carinii pneumonia (PCP) in the absence of cytotoxic or corticosteroid therapy. Gliomas are known to cause immunosuppression, and PCP prophylaxis should be considered for patients with these tumors. PMID- 11881790 TI - Prognostic indicators in cancer: a new approach. PMID- 11881791 TI - Pneumonocystis pneumonia. PMID- 11881792 TI - Neuroblastoma: a single institution's experience with 128 children and an evaluation of clinical and biological prognostic factors. AB - Between 1985 and 1999, 128 children with neuroblastoma were treated at the Royal Alexandra Hospital for Children. The objective of this retrospective study was to report on a single institution's experience with neuroblastoma and to evaluate clinical and biological prognostic factors using univariate and multivariate assessment. Fifty-two percent presented with localized disease, 41% had stage IV disease, and 7% had stage IVs disease. The 5-year overall survival rate was 65%. Significant prognostic factors in univariate analysis included stage, site, histology, N-myc amplification, neuron-specific enolase (NSE), lactate dehydrogenase (LDH), and urinary dopamine. In multivariate assessment, the adjusted hazard ratio was 3.5 (95 %CI 1.4-8.5) for N-myc amplification, 8.7 (95% CI 3.0-25) for NSE > 300 ng/mL, and 3.6 (95% CI 1.3-10) for-LDH > 3000 U/L. This study confirmed that survival was heavily influenced by closely interrelated clinical and biological factors. Prospective studies including more recently identified molecular prognostic factors are warranted to predict the biological heterogeneity of neuroblastoma. PMID- 11881793 TI - Hematological reference ranges in black very low birth weight infants. AB - This study compared hematological reference ranges in black very low birth weight infants to previously published values established predominantly on white subjects. Ninety-four healthy, black, premature babies with a birth weight of 800 to 1500 g at 2-7 days of age were enrolled as part of a study comparing blood transfusions and high- versus low-dose recombinant erythropoietin in anaemia of prematurity. Peripheral venous blood was collected for a full blood count and differential, fetal hemoglobin and erythropoietin levels. The hematological parameters observed in black very low birth weight neonates are similar to previously published reference ranges, except that lower limits of normal were observed for hemoglobin and the red cell indices. PMID- 11881794 TI - Reduction in transfusion requirements with early epoetin alfa treatment in pediatric patients with solid tumors: a case-control study. AB - In a single-center, case-control study the authors evaluated the efficacy and safety of epoetin alfa in pediatric cancer patients receiving platinum- or nonplatinum-based chemotherapy. Thirty-seven patients with solid tumors received epoetin alfa 3 times weekly at a dose of 150 IU/kg (hemoglobin [Hb] > or = 12 g/dL and < or = 16 g/dL) or 300 IU/kg (Hb) < 12 g/dL) for 28 weeks. Data from treated patients were compared to data from 37 untreated control patients. Significant between-group differences in favor of the epoetin alfa-treated Patients were observed in overall red blood cell transfusion requirements (p = .007) and overall platelet transfusion requirements (p = .010). Additionally, significant between-group differences favoring epoetin alfa were seen by Kaplan Meier plots, estimating mean time to first red blood cell transfusion (p = .0004). Mean Hb (g/dL) was maintained at baseline levels in the epoetin alfa group for most of the course of the study. No drug-related adverse events were seen in epoetin alfa-treated patients. PMID- 11881795 TI - Measurements in potassium-supplemented athletes during and after hypokinetic and ambulatory conditions. AB - Hypokinesia (diminished movement) induces significant potassium (K) changes; however, little is known about K deposition and deficiency during hypokinesia (HK). Using K supplements during and after HK, the aim was to establish body K deposition and K deficiency during HK. Studies were done during the pre-HK period of 30 d, HK period of 364 d, and post-HK period of 30 d. Forty male trained athletes aged 24.9 +/- 8.0 y were chosen as subjects. They were equally divided into four groups: unsupplemented active control subjects (UACS), unsupplemented hypokinetic subjects (UHKS), supplemented active control subjects (SACS), and supplemented hypokinetic subjects (SHKS). Hypokinetic subjects were limited to an average walking distance of 0.7 km/d. Control subjects ran an average distance of 11.6 km/d. The SHKS and SACS groups took 95.0 mg elemental K/kg body weight daily. Fecal K excretion, urinary sodium (Na) and K excretion, plasma K and Na levels, plasma renin activity (PRA), plasma aldosterone (PA), food and fluid intake, and physical characteristics were measured. During HK, fecal K loss, urinary K and Na loss, and plasma K, Na, PRA, and PA levels increased significantly (p < or = 0.05), whereas during the initial days of post-HK, the levels of the measured parameters decreased significantly (p < or = 0.05) in the SHKS and UHKS groups as compared with the SACS and UACS groups, respectively. During HK, body weight, body fat, peak oxygen uptake, food and fluid intake decreased significantly (p < or = 0.05), whereas during the initial days of post HK period remained significantly (p < or = 0.05) depressed and fluid intake increased in SHKS and UHKS groups when compared with the SACS and UACS groups, respectively. However, during HK and post-HK plasma, urinary, and fecal K changed significantly (p < or = 0.05) more in the SHKS group than in the UHKS group. The deposition of K was significantly (p < or = 0.05) lower and K deficiency much higher in the SHKS group than in the UHKS group. Fecal K loss, urinary K and Na loss, plasma K, Na, PRA, and PA levels, body weight, body fat, peak oxygen uptake, and food and fluid intake did not change significantly in the SACS and UACS when compared with their baseline control values. It was shown that plasma K concentration and urinary and fecal K excretion increased during HK and decreased significantly (p < or = 0.05) during post-HK. Oral K supplements did not influence plasma or fecal and urinary K either during HK or post-HK. It was concluded that the low plasma K level and fecal and urinary K loss during post-HK may indicate the presence of K deficiency, and increased K in plasma, urine, and feces during HK and in the presence of K deficiency may suggest the body's inability to retain K during HK. PMID- 11881796 TI - Action of boron at the molecular level: effects on transcription and translation in an acellular system. AB - It has been shown that boric acid has well-defined biological effects such as stimulation of wound healing in vivo, release of growth factors and cytokines, and increase of the extracellular matrice turnover. We examined its action at the molecular level, using cell-free systems of transcription (isolated placenta nuclei) and translation (wheat germ extract). We found that 10 mM boric acid greatly increased RNA synthesis, measured by absorbance at 260 nm (x 6.4) or by [3H]-UTP uptake (x 11). Full-length functional mRNA was produced because proteins of 14-80 kDa were translated. Among these proteins, factors involved in angiogenesis and, subsequently, in wound healing (VEGF and TGFbeta) were identified by slot blot, whereas growth factors such as FGF1 and TNFalpha were not detected. These results demonstrate that boron may contribute to biological cell activities at both the transcription and translation levels. However, the mechanism of action is still not known. PMID- 11881797 TI - Effects of vitamin C and vitamin E on lipid peroxidation, blood serum metabolites, and mineral concentrations of laying hens reared at high ambient temperature. AB - This study was conducted to determine the effects of vitamin C (L-ascorbic acid) and vitamin E (alpha-tocopherol acetate) on serum concentrations of lipid peroxidation (MDA) and triiodothyronine (T3), thyroxine (T4), adrenocorticotropic hormone (ACTH), and some metabolite and mineral in laying hens reared at high ambient temperatures ranging from 25 degrees C to 35 degrees C. One hundred twenty laying hens (18 wk old; Hy-Line) were divided into 4 groups, 30 hens per group. The laying hens were fed either a basal diet (control) or the basal diet supplemented with either 250 mg of L-ascorbic acid/kg of diet (vitamin C), 250 mg of alpha-tocopherol acetate/kg of diet (vitamin E), or 250 mg of L-ascorbic acid plus 250 mg alpha-tocopherol acetate/kg of diet (combination). Separately or as a combination vitamins C and E increased serum vitamin C and vitamin E concentrations (p < 0.001) but decreased serum MDA concentration (p < 0.05). Serum concentrations of vitamin E and vitamin C were found highest but serum MDA concentration was lowest in the combination group. Supplemental vitamins C and E either separately or in a combination increased serum T3 and T4 concentrations (p < 0.05), whereas decreased serum ACTH concentration (p < 0.01). Serum glucose and cholesterol concentrations decreased, whereas serum protein concentration increased (p < 0.05) when vitamins C and E singly or together were added to the diet. Vitamin C and vitamin E supplementation resulted in an increase in serum concentrations of Ca, P, and K (p < 0.01) but a decrease in serum concentration of Na (p < 0.05). The results of the present study suggest that supplemental vitamin C and vitamin E alter serum lipid peroxidation, vitamin C, vitamin E and metabolite status, and diets supplemented with a combination of these two vitamins offer a good management practice in laying hens reared at high temperatures. In addition, the results suggest that dietary vitamin C and vitamin E act synergistically. PMID- 11881798 TI - Effects of dietary chromium and zinc on egg production, egg quality, and some blood metabolites of laying hens reared under low ambient temperature. AB - This experiment was conducted to evaluate the effects of chromium (chromium picolinate, CrPic) and zinc (ZnSO4 x H2O) on egg production, egg quality, and serum insulin, corticosterone, glucose, cholesterol, and total protein concentrations of laying hens reared under a low ambient temperature (6.8 degrees C). One hundred twenty laying hens (Hy-Line; 32 wk old) were divided into 4 groups, 30 hens per group. The laying hens were fed the control diet (T1) or the control diet supplemented with either 400 microg of Cr/kg of diet (T2), 30 mg of Zn/kg of diet (T3), or 400 microg of Cr plus 30 mg of Zn/kg of diet (T4). Although the dry matter intake (DMI) was similar (p > 0.05) for all treatment groups, supplemental chromium and zinc either individually or together increased live-weight change, egg production, and improved feed efficiency (p < 0.05). However, no significant differences were observed between T4 and T2 or T3. Compared to T1, supplemental chromium and zinc increased egg weight, eggshell weight, eggshell thickness, egg specific gravity, and Haugh unit (p < 0.05) in T2, T3, and T4 groups, among which there was no significant difference. Serum insulin concentration increased (p < 0.05) and corticosterone concentration decreased (p < 0.05) with dietary chromium and zinc supplementation. Serum glucose and choles- terol concentrations decreased (p < 0.05) and protein concentrations increased (p < 0.001) with dietary chromium and zinc supplementation in all treatment groups. The results of this study indicated that either supplemental dietary chromium or zinc increased plasma insulin and decreased corticosterone concentrations and that had a positive effect on performance of laying hens under low ambient temperature. PMID- 11881799 TI - Protective role of supplemental vitamin E and selenium on lipid peroxidation, vitamin E, vitamin A, and some mineral concentrations of Japanese quails reared under heat stress. AB - This study was conducted to determine the effects of vitamin E and selenium (Se) on lipid peroxidation (MDA), serum and liver concentration of antioxidant vitamins, and some minerals of Japanese quails reared under heat stress (34 degrees C). One hundred twenty 10-d-old Japanese qualis (60 males, 60 females) were randomly assigned to 4 treatment groups, 3 replicates of 10 birds each. The experiment was designed in a 2x2 factorial arrangement using two levels of vitamin E (125 and 250 mg/kg of diet) and two levels of selenium (0.1 and 0.2 mg/kg of diet). Greater dietary vitamin E and selenium inclusions resulted in a greater (p = 0.001) serum vitamin E and vitamin A, but lower (p = 0.001) MDA concentrations. Liver vitamin E and vitamin A concentrations increased (p = 0.001) and MDA concentrations decreased (p = 0.001) when both dietary vitamin E and selenium increased. No interactions between vitamin E and selenium were detected (p > or = 0.11) for any parameters. Increasing both dietary vitamin E and selenium caused an increase in serum concentrations of Fe and Zn (p = 0.001), but a decrease in serum concentration of Cu (p = 0.001). Results of the present study showed that dietary vitamin E and selenium have synergistic effects and that supplementing a combination of dietary vitamin E (250 mg/kg of diet) and selenium (0.2 mg/kg of diet) offers a good management practice to reduce heat stress-related depression in performance of Japanese quails. PMID- 11881800 TI - Malnutrition interferes with the unique lymphadenopathy induced in Lewis rats by metallic tin. AB - Metallic tin powder injected into Lewis rats caused marked enlargement of draining lymph nodes with prominent hyperplasia of plasma cells. A low level of dietary protein (8%) or deficiency of choline interfered with the cellular proliferation and reduced the size of the nodes by nearly one-third compared to a normal protein diet (20%). A high level of protein in the diet (50%) did not augment the effect of metallic tin powder beyond that achieved by a level usually considered adequate (20%). PMID- 11881801 TI - Uptake and distribution of trace metal elements in wheat seedlings. AB - The uptake and distribution of eight metallic elements were examined in wheat seedlings for a period of 12 d with a radioactive multitracer technique. The radioactive nuclides of the seedlings were simultaneously determined by gamma-ray spectrometry. All of the elements studied were taken up by the wheat seedlings and mainly accumulated in the roots. Only some elements were transported to shoots and leaves of the seedlings or bound to leaf proteins, and two elements were transported into the chloroplast. Uptake of most elements reached a maximum on the fifth or the eighth day and then gradually decreased afterward. In the cases of 95mTc and 72Se, the uptake increased continuously within 12 d without the peak uptake. The change of elemental concentrations was dependent on uptake and excretion rates. The dynamics of metal elements taken up by the wheat seedlings and their distribution in roots, shoots, and leaves were different for each element, suggesting that it may depend on the characteristics of the elements. PMID- 11881802 TI - Colorimetric determination of chloride in biological samples by using mercuric nitrate and diphenylcarbazone. AB - A colorimetic method is outlined for the determination of the chloride ion in biological samples (blood serum, plasma, and urine). The present method is based on the quantitative reduction of free mercuric ions by chloride ions. Chloride ions form an indissociable complex with mercuric ions. The remaining free mercuric ions form a purple complex with diphenylcarbazone with an absorption maximum at 550 nm. The reduction of color intensity at 550 nm is directly proportional to chloride concentration in the sample. The linear concentration range in the final reaction mixture was 0-100 microM with a correlation coefficient of -0.9997. The coefficient of variation for the 50 microM chloride ion in the final reaction mixture was 0.9% (n = 6). The analyzed value of chloride concentration in the human control serum Accutrol Normal (Sigma) was 101+/-4 mM (mean+/-SD, n = 12). The certified value of chloride in Accutrol Normal by Sigma is 102 mM, with a mean in the range 91-113 mM. This method was applied to the measurement of urinary chloride excretion in experimental rats. During 16-h urine collection, no food was given and rats had free access to purified water. The urinary excretion rate of chloride was 23.6+/-9.3 micromol/h (mean+/-SD, n = 8) and 126.2+/-28.0 micromol/h (n = 8) for rats fed a normal diet (2.6 g NaCl/kg diet) and a high-salt diet (82.6 g NaCl/kg diet) for 70 d prior to urine collection, respectively. This method is appropriate for low concentrations of chloride in samples or when sample volume is limiting, as in many animal studies such as metabolic urine collection from rats. PMID- 11881803 TI - Aedes aegypti transducing densovirus pathogenesis and expression in Aedes aegypti and Anopheles gambiae larvae. AB - Aedes aegypti densovirus (AeDNV) is a small DNA virus that has been developed into an expression and transducing vector for mosquitoes [Afanasiev et al. (1994) Exp Parasitol 79: 322-339; Afanasiev et al. (1999) Virology 257: 62-72; Carlson et al. (2000) Insect Transgenesis: Methods and Applications (Handler, A.M. & James, A.A., eds), pp. 139-159. CRC Press, Boca Raton]. Virions carrying a recombinant genome expressing the GFP gene were used to characterize the pathogenesis of the virus in 255 individual Aedes aegypti larvae. The anal papillae of the larvae were the primary site of infection confirming previous observations (Afanasiev etal., 1999; Allen-Muira et al. (1999) Virology 257: 54 61). GFP expression was observed in most cases to spread from the anal papillae to cells of the fat body, and subsequently to many other tissues including muscle fibers and nerves. Infected anal papillae were also observed to shrink, or melanize and subsequently fall off in a virus dependent manner. Three to four day old larvae were less susceptible to viral infection and, if infected, were more likely to survive into adulthood, with 14% of them still expressing GFP as adults. Higher salt concentrations of 0.10-0.15 M inhibited viral infection. Anopheles gambiae larvae also showed infection of the anal papillae (17%) but subsequent viral dissemination did not occur. The persistence of the reporter gene expression into adulthood of Aedes aegypti indicates that transduction of mosquito larvae with recombinant AeDNV may be a means of introducing a gene of interest into a mosquito population for transient expression. PMID- 11881804 TI - Ribosomal DNA spacer genotypes of the Anopheles bancroftii group (Diptera: Culicidae) from Australia and Papua New Guinea. AB - Mosquitoes of the Anopheles bancroftii group collected from Northern Australia and Papua New Guinea (PNG) were investigated for sequence variation within the ribosomal DNA ITS2. Wing fringe morphology originally used to identify members of this group was compared to genotypes identified by restriction fragment length polymorphism analysis (RFLP) and heteroduplex analysis (HDA) of the rDNA ITS2. Members of this group separated into four RFLP genotypes (A, B, C and D) with some genotypes displaying wing fringe polymorphisms. Heteroduplex analysis of the ITS2 within and between populations identified genotype A as containing two geographically separate ITS2 sequences: A1 from the Northern Territory of Australia and A2 from Queensland and the Western Province of PNG. Genotypes B and C and genotypes C and D were found sympatric and appeared to be evolving independently suggesting the possibility of cryptic species. Genotype C contained two ITS2 sequence types within the genome. PMID- 11881806 TI - MsqTc3, a Tc3-like transposon in the yellow fever mosquito Aedes aegypti. AB - A novel transposon, MsqTc3-Aa, has been discovered in the yellow fever mosquito, Aedes aegypti. Evidence of its past mobility is presented. There are approximately 100 copies of MsqTc3-Aa in A. aegypti, eight of which have been isolated and sequenced. All sequenced copies are more than 99% identical to their consensus, indicating recent mobilization. The MsqTc3-Aa consensus contains imperfect terminal inverted repeats (TIRs) and an open reading frame (ORF) interrupted by an intron. Sequence, structural and phylogenetic analysis showed that MsqTc3-Aa is a distant relative of Tc3, an active transposon in Caenorhabditis elegans. These results may provide useful information for the current effort to control mosquito-borne diseases using genetic approaches. PMID- 11881805 TI - The internal transcribed spacers (ITS1 and ITS2) of the rDNA differentiates the bark beetle forest pests Tomicus destruens and T. piniperda. AB - The bark beetles species Tomicus destruens and T. piniperda constitute one of the main pests of European and Asian forests. T. destruens has been thought to be the same species as T. piniperda by some authors. A rapid PCR-based method using internal transcribed spacers (ITS1 and ITS2) of the ribosomal DNA has been developed in order to identifying these two species. We have studied differences in the size of the ITS1, and differences in the sequence and restriction patterns of the ITS2 in 75 individuals of both species. The results indicate a size difference of about 100 bp in the ITS1 of T. destruens and T. piniperda. The size of the ITS2 spacer is similar in both species (approximately 600 bp). However, the sequence is consistently different between T. destruens and T. piniperda in all populations analysed, showing some characteristic indels. Differences in the restriction target in both species for the enzymes HincII and DraI produce different band patterns in agarose NuSieve 5% gel electrophoresis. These data suggest the validity of T. destruens as a phylogenetically differentiated entity from T. piniperda. The phylogenetic analysis of a 789 bp fragment of the 3' end of the cytochrome oxidase I gene (COI) agrees with the two groupings obtained with the ITS2, showing a phylogenetic clustering rather than a phylogeographic structure. PMID- 11881807 TI - A multiplex PCR-based method derived from random amplified polymorphic DNA (RAPD) markers for the identification of species of the Anopheles minimus group in Southeast Asia. AB - Effective control of Anopheles minimus s.l., an important malaria vector in Southeast Asia, is based on the accurate identification of species within An. minimus complex, which cannot be distinguished using morphological characters. Derived from individual random amplified polymorphic DNA markers, sequence characterized amplified regions were analysed for the design of species-specific paired-primers. Combination of these primers resulted in the development of a simple, robust multiplex PCR able to identify both species An. minimus A and C belonging to the complex, hybrids AC, and three sympatric and closely related species, An. aconitus, An. pampanai and An. varuna. Hybrids AC do not possess alleles of both parents but exhibit novel adaptive potentials resulting from recombination among parental genes leading to hybrizyme. PMID- 11881808 TI - Identification of four small molecular mass proteins in the silk of Bombyx mori. AB - This paper describes cDNAs of four small-size proteins that occur in the cocoon silk of Bombyxmori. Two of them (9.9 and 10.3 kDa), which have the N-terminal sequences and the spacing of a few amino acids at C-termini similar to the seroin of Galleria mellonella, are called seroin 1 and seroin 2. The corresponding genes are expressed in the middle, and to a small extent also in the posterior silk gland sections. The seroin 1 and less conspicuously the seroin 2 mRNAs accumulate in the course of the last larval instar to a maximum in postspinning larvae. Two other proteins (6 kDa and 4.7 kDa) of B. mori cocoons were identified as a typical Kunitz-type and a somewhat unusual Kazal-type proteinase inhibitors, and named BmSPI 1 and BmSPI 2, respectively. Their genes are expressed in the middle, and the BmSPI 1 gene slightly also in the posterior silk gland sections. The expression ensues a few days after the last larval ecdysis and increases until the cocoon spinning. Post-spinning larvae still contain high amounts of the BmSPI 1 but no BmSPI 2 transcripts. It is assumed that seroins and proteinase inhibitors are involved in cocoon protection against predators and microbes. PMID- 11881809 TI - piggyBac-mediated transposition in Drosophila melanogaster: an evaluation of the use of constitutive promoters to control transposase gene expression. AB - Transgenic non-Drosophilid insects have been made using insect transposable elements that have a broad host range such as the piggyBac element. However, the success rate is often low. Previous piggyBac helper plasmids have used either the piggyBac or the hsp70 promoter from Drosophila melanogaster to control expression of the transposase gene. Here we show that plasmids with the piggyBac transposase gene regulated by constitutive promoters can be effective 'helpers' for mediating transposition in D. melanogaster. We also present preliminary evidence on the use of an RNA helper that encodes the transposase. Our results suggest that for germ line transformation of non-Drosophilid insects it may be advantageous to isolate a constitutive promoter from the species of interest to control transposase expression. PMID- 11881810 TI - Tehao functions in the Toll pathway in Drosophila melanogaster: possible roles in development and innate immunity. AB - Toll and related proteins play important roles in innate immunity in both invertebrates and vertebrates. In Drosophila melanogaster, Tehao shares a striking similarity in its intracellular domain with Toll. In this paper, we show that Tehao is expressed throughout development and appears to be glycosylated. In transiently transfected cells, Tehao activated both Dorsal and the transcription of endogenous drosomycin and metchnikowin genes. Purified recombinant Tehao interacted specifically in vitro not only with the Pelle protein kinase, but also with the Toll intracytoplasmic domain. Remarkably, Tehao was found to activate Dorsal-dependent transcription in a synergistic manner with Toll, as well as Pelle in co-transfected cells. Thus, Tehao, alone or with Toll as a multimeric complex, has the potential to participate in both the development and innate immune responses of Drosophila. PMID- 11881811 TI - Organization and developmental expression of the mosquito vitellogenin receptor gene. AB - Vitellogenin is a precursor of the major yolk protein, vitellin. It is internalized by developing oocytes via receptor-mediated endocytosis. Previously, we characterized the vitellogenin receptor (VgR) from oocytes of the mosquito Aedes aegypti [Sappington, T.W., Kokoza,V.A., Cho,W.L. and Raikhel,A.S. (1996) Molecular characterization of the mosquito vitellogenin receptor reveals unexpected high homology to the Drosophila yolk protein receptor. Proc Natl Acad Sci USA 93: 8934-8939]. The VgR receptor has a unique structure with two putative ligand-binding domains. In order to understand the regulation of this important molecule, we characterized the VgR gene structure and its expression during vitellogenesis in the mosquito A. aegypti. We report here that the VgR gene was separated by five introns that have an average length of 60 bp, except for the second intron which was more than 20 kb long. Most introns were located within the coding regions of the first protein domain. We isolated two allelic variations of the VgR gene, VgR1 and VgR2, the nucleotide sequences of which differing only in their 5'-flanking regions. Considering their frequency in the mosquito genome, VgR2 appeared to be a major allele. The expression of VgR mRNA was studied by the Northern blot analysis and in situ hybridization. The level of the VgR transcript started to rise in the ovary one day post-eclosion. It continued its dramatic rise during the vitellogenic period, reaching its peak at 24 h PBM. The VgR transcript was present exclusively in ovaries where it was seen in oocytes and nurse cells of primary follicles and germ-line cells of the germarium. PMID- 11881812 TI - Molecular phylogeny of Calyptratae (Diptera: Brachycera): the evolution of 18S and 16S ribosomal rDNAs in higher dipterans and their use in phylogenetic inference. AB - Sequences for nearly complete 18S rRNA and partial 16S rRNA genes were determined for sixteen species representing twelve calyptrate families. Two unique insertions are present in expansion regions of the 18S rDNA in nycteribiids. Alignments containing other dipteran rRNA genes provided good resolution at higher taxonomic level: monophyly of Calyptratae is well supported. While both 16S and 18S rDNA matrices produce unstable topologies within Calyptratae when analysed separately, their combination results in a tree with several robust and well supported nodes. Of three superfamilies recognized in recent classifications, the Hippoboscoidea is well supported by 16S rDNA and by combined matrices. The representatives of Muscoidea, Musca sp. and Antipoda sp., display a tendency to cluster within Oestroidea. The comparison of secondary structures of two variable regions indicates that Sarcophagidae are related to Calliphoridae rather than to Tachinidae. PMID- 11881813 TI - Identification of a novel gene, Mblk-1, that encodes a putative transcription factor expressed preferentially in the large-type Kenyon cells of the honeybee brain. AB - Mushroom bodies (MBs) are considered to be involved in higher-order sensory processing in the insect brain. To identify the genes involved in the intrinsic function of the honeybee MBs, we searched for genes preferentially expressed therein, using the differential display method. Here we report a novel gene encoding a putative transcription factor (Mblk-1) expressed preferentially in one of two types of intrinsic MB neurones, the large-type Kenyon cells, which makes Mblk-1 a candidate gene involved in the advanced behaviours of honeybees. A putative DNA binding motif of Mblk-1 had significant sequence homology with those encoded by genes from various animal species, suggesting that the functions of these proteins in neural cells are conserved among the animal kingdom. PMID- 11881814 TI - The thymidylate synthase gene of Hz-1 virus: a gene captured from its lepidopteran host. AB - The sequence analysis of a thymidylate synthase gene was identified in the Hz-1 virus HindIII-D fragment. The viral thymidylate synthase gene encodes a protein of 295 amino acids, and is closely related to that of insect, mammals and herpesvirus. The thymidylate synthase gene identified was a genuine viral gene in that it was only detected in cells infected with Hz-1 virus but not in the mock infected cells, by Southern blot analysis and by RT-PCR. Results of phylogenetic analysis based on non-synonymous and amino acid distances suggested that the TS gene of Hz-1 virus was grouped closely with that of Bombyx mori. High bootstrap values confirmed that the thymidylate synthase of Hz-1 virus was acquired by a capture event from its lepidopteran host. Results of both sequence divergences and phylogenetic analysis suggested that TS genes in insect viruses, Hz-1, CIV, and MsEPV may have a different history or originated from different capture events. PMID- 11881815 TI - Characterization of a cDNA encoding a cathepsin L-like protein of Rhodnius prolixus. AB - The interaction of Rhodnius prolixus digestive enzymes with Trypanosoma cruzi could be important for parasite survival. We report herein the complete sequence of the messenger of a cathepsin L-like molecule (RpCat). The cDNA has 5'- and 3'- end UTRs and a methionine codon that corresponds likely to a translation initiation codon. In the deduced amino acid sequence, a region corresponding to an ERFININ domain, diagnostic of L-cathepsins, and a possible pro-peptide cleavage site were observed. At the C-terminus, a nine-amino acid sequence, almost identical to a secretion signal of human cathepsin L was found. RpCat messenger was expressed in intestines of R. prolixus adults, and from 1st to 4th but not in 5th instar nymph stages. In a similarity analysis, RpCat was grouped with L cathepsins forming a clear group separate of the B cathepsins. PMID- 11881816 TI - A molecular study of abdominal-A in the ant Myrmica rubra reveals lineage dependent evolutionary rates for a developmental gene. AB - We have characterized the abdominal-A locus in a Hymenopteran, the ant Myrmica rubra. The sequence of the homeotic domain of the Abdominal-A family of orthologous genes is known for a fairly large number of insects but the complete sequence of the Abdominal-A protein, is known only for a few. The two proteins of Drosophila melanogaster and Tribolium castaneum differ markedly outside the homeodomain. A comparison of the ant Abdominal-A protein sequences with those of these two insects shows that the ant and beetle sequences are very similar all along the length of the protein. The fruit fly has diverged considerably and equally from the other two insects. This divergence reflects different rates of evolution of the protein in different lineages. PMID- 11881817 TI - Minor H antigens: genes and peptides. AB - In this review, we describe the evidence from which the existence of non-MHC histocompatibility (H) antigens was deduced, the clinical setting of bone marrow transplantation in which they are important targets for T-cell responses, and the current understanding of their molecular identity. We list the peptide epitopes of the human and murine minor H antigens now identified at the molecular level, their MHC restriction molecules and the genes encoding them. Identification of the peptide epitopes allows T-cell responses to these antigens following transplantation of MHC-matched, minor H-mismatched tissues to be enumerated using tetramers and elispot assays. This will facilitate analysis of correlations with host-versus-graft (HVG), graft-versus-host (GVH) and graft-versus-leukaemia (GVL) reactions in vivo. The potential to use minor H peptides to modulate in vivo responses to minor H antigens is discussed. Factors controlling immunodominance of T-cell responses to one or a few of many potential minor H antigens remain to be elucidated but are important for making predictions of in vivo HVG, GVH and GVL responses and tailoring therapy after HLA-matched bone marrow transplantation and donor lymphocyte infusion. PMID- 11881818 TI - The human complement C9 gene: structural analysis of the 5' gene region and genetic polymorphism studies. AB - C9 is the last of the human complement components creating the membrane attack complex. The single chain serum protein is encoded by a gene located on chromosome 5p13 that is composed of 11 exons. With the aid of inverse PCR, the hitherto unknown regions flanking exon 1 and the 3' part of exon 11 (3'UTR) have been sequenced. A computer-based analysis of the 300-bp region located just upstream of the AUG start codon showed homologies to known DNA modules which affect the transcriptional regulation of certain genes. The most striking of these is a sequence that may substitute the missing TATA box in initiating C9 transcription. In the 3'UTR, three successive polyadenylation signals were found. Although the C9 protein is invariant, four different single nucleotide polymorphisms (SNPs) have been observed at the DNA level by exon-specific PCR and direct sequencing. None of them changes the amino acid composition of the mature protein. Due to a C --> T transition in exon 1 at cDNA position 17, the fifth amino acid of the leader peptide may be either an arginine or a tryptophane. Using either PCR/ RFLP analysis (exons 1 and 11) or allele-specific PCR (intron 1 and exon 4), each polymorphism can be characterized without sequencing. All of the exon 1, intron 1 and exon 11 variants could be detected in small population samples of European, Thai or South American Indian origin. In contrast, the exon 4 C variant was observed only once in a European. The first three SNPs can be combined to designate eight different 'C9 alleles'. Of these, six have actually be found. These data provide strong evidence that several mutation and recombination events occurred in the course of C9 gene evolution. PMID- 11881819 TI - MICA and MICB microsatellite alleles in HLA extended haplotypes. AB - The present study is a contribution to the definition of the linkage disequilibrium relationship of MICA and MICB with adjacent loci and to the characterization of extended HLA haplotypes. These issues are of importance for the identification of disease associations and for a better definition of donor recipient compatibility in bone-marrow grafts through the typing of haplospecific markers. The distribution of the five alleles of MICA and the 13 alleles of MICB microsatellites, located, respectively, in MICA transmembrane exon 5 and in MICB intron 1, was examined in 133 healthy Italian individuals previously typed for HLA class I, class II and complement loci and for the TNFa microsatellite. The MICB microsatellite was also analysed in 49 HTCLs for which MICA typing was already available. Very strong linkage disequilibria with HLA-B and TNFa were detected in the Italian population for both MICA and MICB microsatellite alleles, in spite of the high mutability rate of the larger MICB alleles. Some strong associations were also detected between MICB and DRB1. The strongest associations (P < 0.001, D' > 0.7) were those of MICA-A4 with HLA-B18, B27 and TNFa1, MICA-A5 with HLA-B35, B61 and B62, MICA-A5.1 with HLA-B7, B8, B13, B63 and MICB-CA24, MICA-A6 with HLA-B51, MICA-A9 with HLA-B39, B57 and TNFa2, MICB-CA14 with HLA B14, B27 and TNFa1, MICB-CA15 with HLA-B52, TNFa4 and TNFa13, MICB-CA17 with HLA B7 and TNFa11, MICB-CA18 with HLA-B13 and TNFa7, MICB-CA22 with HLA-B57, and MICB CA24 with HLA-B8 and TNFa2. From pairwise associations in the random panel and results for the homozygous cell lines it was possible to deduce the MICA and MICB microsatellite alleles present in many of the well-known Caucasoid extended haplotypes. PMID- 11881820 TI - A study of Gm allotypes and immunoglobulin heavy gamma IGHG genes in Berbers, Arabs and sub-Saharan Africans from Jerba Island, Tunisia. AB - The Gm polymorphism of human IgG immunoglobulins was investigated in three different ethnic groups--Arabs, Berbers and 'dark-skinned people'--on Jerba Island, Tunisia. The genetic relationships among these groups and several populations from North Africa, sub-Saharan Africa, west Asia and Europe were analysed by principal coordinate analysis, Fst significance testing, and analysis of molecular variance based on haplotype frequencies. The results revealed a non significant genetic differentiation between Arabs and Berbers from Jerba. However, the Jerbian population of sub-Saharan African origin was close to Ethiopians. Gene flow among the three Jerbian populations, as well as an East African origin of the dark-skinned individuals, is proposed to account for the observed genetic pattern. However, the genetic diversity observed among the different Tunisian populations did not show any significant correlation with either geographic or linguistic differentiation. A preliminary analysis of the restriction fragment length polymorphism of the IGHG genes in Arabs and Berbers from Jerba confirmed the close genetic relationship between the two populations. However, it also indicated a lower level of genetic diversity in the Berbers, which may be explained by more rapid genetic drift due to longer isolation on the island. PMID- 11881821 TI - HLA DMA and DMB show no association with rheumatoid arthritis in US Caucasians. AB - HLA DM is a heterodimeric molecule functioning in normal antigen presentation; it is encoded by adjacent HLA-region loci, HLA DMA and DMB, located between DP and DQ. Some previous studies have suggested that HLA susceptibility to rheumatoid arthritis (RA) is associated with certain DMA and DMB alleles. Our aim was to examine whether this association is also present in US Caucasians. We studied 288 US Caucasian subjects with rheumatoid arthritis and 263 US Caucasian control subjects. DMA and DMB typing was achieved by PCR amplification followed by sequence-specific oligonucleotide hybridization and by PCR-restriction fragment length polymorphism. There was no frequency difference for DMA alleles or DMB alleles between RA and control subjects, indicating no association. Neither was a difference apparent when data were analysed in subgroups based on shared-epitope DRB1, on the rheumatoid factor test, on radiographic changes of RA, or on sex. DRB1-DQB1-DMB analyses for linkage disequilibrium showed that the DRB1*0401 DQB1*0301 haplotype had the DMB*0103 allele more often than DMB*0101 (estimated haplotype frequencies 0.08 and 0.039 in RA, respectively). In contrast, the DRB1 *0401-DQB1 *0302 haplotype usually had the DMB*0101 allele (haplotype frequency 0.084 compared to 0.01 for DMB*0103). Thus, neither HLA DMA nor DMB was associated with RA in this population, and not all shared-epitope-bearing haplotypes had the same DMB allele distribution. PMID- 11881822 TI - Short-form HLA-DP typing with 48 primer mixes using the polymerase chain reaction and sequence-specific primers. AB - We have developed a short-form SSP-based HLA-DP typing system for routine use adapted from a comprehensive HLA-DP typing method described by Gilchrist et at. (1998). Our short-form system detects 93 alleles, including the 18 most common HLA-DPB1 alleles and eight HLA-DPA1 alleles. The primer mixes described were tested using the PCR-SSP Manager (Bunce et al., 1998) database to confirm the specificity of selected primers, and to detect potentially ambiguous amplifications. This short-form HLA-DP typing system was validated using 50 fully typed samples obtained through the UCLA International DNA Exchange. All samples gave 100% concordance with the consensus type. Our laboratory routinely uses a PCR-SSP based system of 48 primer mixes for HLA-DRB and HLA-DQB typing. The advantage of the short-form HLA-DP typing system described here is that the additional 48 HLA-DP primer mixes required can be included on the second half of a 96-well format tray. This method now enables a full HLA class II typing at the level of allele group resolution in 2 1/2 h. PMID- 11881823 TI - Identification of a polymorphic microsatellite marker in the mouse Fas ligand gene. AB - We describe the characterization of a polymorphic microsatellite marker, located 1.8 kb downstream of exon 4 in the mouse Fas ligand gene. This (GT) repeat sequence allows the identification of four alleles which can very easily be distinguished by simple agarose electrophoresis. PMID- 11881824 TI - Nomenclature for factors of the HLA system, update April 2001. PMID- 11881825 TI - Nomenclature for factors of the HLA system, update May 2001. PMID- 11881826 TI - Nomenclature for factors of the HLA system, update June 2001. PMID- 11881827 TI - A novel PCR-rFLP assay for the detection of the single nucleotide polymorphism at position -1082 in the human IL-10 gene promoter. PMID- 11881828 TI - Physical mapping, BAC-end sequence analysis, and marker tagging of the soilborne nematicidal bacterium, Pseudomonas synxantha BG33R. AB - A bacterial artificial chromosome (BAC) library was constructed for the genome of the rhizosphere-inhabiting fluorescent pseudomonad Pseudomonas synxantha BG33R. Three thousand BAC clones with an average insert size of 140 kbp and representing a 70-fold genomic coverage were generated and arrayed onto nylon membranes. EcoRI fingerprint analysis of 986 BAC clones generated 23 contigs and 75 singletons. Hybridization analysis allowed us to order the 23 contigs and condense them into a single contig, yielding an estimated genome size of 5.1 Mb for P. synxantha BG33R. A minimum-tile path of 47 BACs was generated and end-sequenced. The genetic loci involved in ring nematode egg-kill factor production in BG33R Tn5 mutants, 246 (vgrG homolog), 1122 (sensor kinase homolog), 1233 (UDP-galactose epimerase homolog), 1397 (ferrisiderophore receptor homolog), and 1917 (ribosomal subunit protein homolog), have been mapped onto the minimum-tile BAC library. Two of the genetic regions that flank Tn5 insertions in BG33R egg-kill-negative mutants 1233 and 1397 are separated by a single BAC clone. Fragments isolated by ligation-mediated PCR of the Tn5 mutagenized regions of 29 randomly selected, non egg-kill-related, insertion mutants have been anchored onto the ordered physical map of P. synxantha. PMID- 11881829 TI - A theoretical limit to coding space in chromosomes of bacteria. AB - A mathematical model of cluster patterns for mapped genes with known phenotypes in Escherichia coli predicted thatfunctional genes may account for a maximum of two-thirds of the total chromosomal space. The corollary prediction was that one third of the chromosome comprised noncoding space. Open reading frame (ORF) analyses for 15 phylogenetically diverse bacterial genomes and for 30 fully sequenced prokaryotic genomes supported the gene cluster model prediction of a two-thirds tendency for coding space. Our results suggest that only 3-4% of unassigned ORFs in E. coli represent genes with potential phenotype and that ORFs marking novel genes in prokaryotes are far fewer than previously thought. PMID- 11881831 TI - 2001 GSAC meeting summary: from genome to transcriptome to proteome. Genome Sequencing and Analysis Conference. PMID- 11881830 TI - Spectral analysis of distributions: finding periodic components in eukaryotic enzyme length data. AB - We introduce the spectral analysis of distributions (SAD), a method for detecting and evaluating possible periodicity in experimental data distributions (histograms) of arbitrary shape. SAD determines whether a given empirical distribution contains a periodic component. We also propose a system of probabilistic mixture distributions to model a histogram consisting of a smooth background together with peaks at periodic intervals, with each peak corresponding to a fixed number of subunits added together. This mixture distribution model allows us to estimate the parameters of the data and to test the statistical significance of the estimated peaks. The analysis is applied to the length distribution of eukaryotic enzymes. PMID- 11881832 TI - The Department of Energy microbial cell project: A 180 degrees paradigm shift for biology. PMID- 11881833 TI - TLFAM--a new set of protein family databases. AB - PFAM is a popular and effective database of Hidden Markov Models (HMMs), which represent a wide range of protein families. Here, we introduce TLFAM as a more specific set of HMM databases. Analyses of bacterial genomes using TLFAM-Pro show better scores, E-values, and alignment lengths than those using the more generalized PFAM. Since PFAM will still find hits that TLFAM-Pro will not, we recommend that they be used jointly, rather than exclusively. This method provides the best features of both databases. This method has been extended to a number of other organism types, such as archaea, and the databases are freely available to interested researchers. PMID- 11881834 TI - Gene and protein expression profiles of Shewanella oneidensis during anaerobic growth with different electron acceptors. AB - Changes in mRNA and protein expression profiles of Shewanella oneidenesis MR-1 during switch from aerobic to fumarate-, Fe(III)-, or nitrate-reducing conditions were examined using DNA microarrays and two-dimensional polyacrylamide gel electrophoresis (2-D PAGE). In response to changes in growth conditions, 121 of the 691 arrayed genes displayed at least a two-fold difference in transcript abundance as determined by microarray analysis. Genes involved in aerobic respiration encoding cytochrome c and d oxidases and TCA cycle enzymes were repressed under anaerobic conditions. Genes induced during anaerobic respiration included those involved in cofactor biosynthesis and assembly (moaACE, ccmHF, nosD, cysG), substrate transport (cysUP, cysTWA, dcuB), and anaerobic energy metabolism (dmsAB, psrC, pshA, hyaABC, hydA). Transcription of genes encoding a periplasmic nitrate reductase (napBHGA), cytochrome c552, and prismane was elevated 8- to 56-fold in response to the presence of nitrate, while cymA, ifcA, and frdA were specifically induced three- to eightfold under fumarate-reducing conditions. The mRNA levels for two oxidoreductase-like genes of unknown function and several cell envelope genes involved in multidrug resistance increased two- to fivefold specifically under Fe(III)-reducing conditions. Analysis of protein expression profiles under aerobic and anaerobic conditions revealed 14 protein spots that showed significant differences in abundance on 2-D gels. Protein identification by mass spectrometry indicated that the expression of prismane, dihydrolipoamide succinyltransferase, and alcaligin siderophore biosynthesis protein correlated with the microarray data. PMID- 11881836 TI - Fifteen minutes of fim: control of type 1 pili expression in E. coli. AB - Pili are used by Escherichia coli to attach to and invade mammalian tissues during host infection and colonization. Expression of type 1 pili, believed to act as virulence factors in urinary tract infections, is under control of the 'firm' genetic network. This network is able to sense the environment and actuate phase variation control. It is a prime exemplar of an integrative regulatory system because of its role in mediating a complex infection process, and because it instantiates a number of regulatory motifs, including DNA inversion and stochastic variation. With the help of a mathematical model, we explore the mechanisms and architecture of the fim network. We explain (1) basic network operation, including the roles of the recombinase and global regulatory protein concentrations, their DNA binding affinities, and their switching rates in observed phase variation behavior; (2) why there are two recombinases when one would seem to suffice; (3) the source of on-to-off switching specificity of FimE; (4) the role of fimE orientational control in switch dynamics; and (5) how temperature tuning of piliation is achieved. In the process, we identify a general regulatory motif that tunes phenotype to an environmental variable, and explain a number of apparent experimental inconsistencies. PMID- 11881837 TI - Naftidrofuryl in the treatment of vascular dementia. AB - The design of this study was based on the European guidelines for the treatment of Alzheimer's disease. After a placebo run-in period of 4 weeks, patients with a diagnosis of vascular dementia (VaD) were randomised to receive either 400 mg naftidrofuryl/day, 600 mg naftidrofuryl/day or placebo for 6 months. The patients were assessed using the ADAS-cog, the SCAG, the NOSGER and the CGI item 2 scale. The primary analysis was undertaken on the ITT population. At the end of the study, significantly more patients in the treatment groups showed no deterioration on both ADAS-cog and SCAG scales compared with placebo (400 mg p = 0.005, 600mg p = 0.015). There were also significant differences between the active and placebo groups for the individual scales. This study has demonstrated that treatment with naftidrofuryl can slow the rate of deterioration of patients with vascular dementia. PMID- 11881835 TI - The use of accurate mass tags for high-throughput microbial proteomics. AB - We describe and review progress towards a global strategy that aims to extend the sensitivity, dynamic range, comprehensiveness, and throughput of proteomic measurements for microbial systems based upon the use of polypeptide accurate mass tags (AMTs) produced by global protein enzymatic digestions. The two-stage strategy exploits high accuracy mass measurements using Fourier transform ion cyclotron resonance mass spectrometry (FTICR) to validate polypeptide AMTs for a specific organism, from potential mass tags tentatively identified using tandem mass spectrometry (MS/MS), providing the basis for subsequent measurements without the need for routine MS/MS. A high-resolution capillary liquid chromatography separation combined with high sensitivity, and high-resolution accurate FTICR measurements is shown to be capable of characterizing polypeptide mixtures of more than 10(5) components, sufficient for broad protein identification using AMTs. Advantages of the approach include the high confidence of protein identification, its broad proteome coverage, and the capability for stable-isotope labeling methods for precise relative protein abundance measurements. The strategy has been initially evaluated using the microorganisms Saccharomyces cerevisiae and Deinococcus radiodurans. Additional developments, including the use of multiplexed-MS/MS capabilities and methods for dynamic range expansion of proteome measurements that promise to further extend the quality of proteomics measurements, are also described. PMID- 11881838 TI - Reliability and reproducibility of brain tissue volumetry from segmented MR scans. AB - Reliable measurement of different tissue volumes in the living brain is of great importance for human brain research. In this article, we report on the inter- and intraoperator reliability and scan-rescan reproducibility of segmented intracranial tissue volumes from MR images using the image analysis software suite BRAINS. The absolute data of tissue volume measurements are also presented. The reliability and consistency of the measurements of the segmented volumes were excellent. The segmentation is robust and rapid and the volume measurements are plausible and suitable for quantitative studies in clinical brain research. PMID- 11881839 TI - Pattern of neuropsychological deficits in patients with treated Wilson's disease. AB - The study aimed to describe the neuropsychological profiles in patients with treated Wilson's disease (WD). The series included 19 symptomatic and 2 asymptomatic patients with a mean age of 35.3 +/- 9.2 years. They were tested with the Automated Psychological Test system (APT), a comprehensive computerised neuropsychological test battery. APT comprised eleven separate tests and assessed five essential types of neuropsychological functions: motor functions, basic neuropsychological functions, specific cognitive functions, memory, and executive functions. The results were compared to current norms of the test battery. The symptomatic WD patients had significantly lower performance than the norms on all finger tapping tasks, the simple reaction time, the simultaneous capacity background task, the short-term memory test, the index of word decoding speed, the grammatical reasoning test, and the perceptual maze test. They were significantly higher on the index of impulsive errors, and used a significantly more global processing mode in the test of selective attention. The female symptomatic patients displayed more pronounced neuropsychological deficits than the males in the complex tasks. WD patients displayed a specific profile of moderate neuropsychological impairment. The results are theoretically interesting and have practical implications for the management of WD patients, e.g. some patients confronted with the results have had increased compliance. PMID- 11881840 TI - Clozapine-associated elevation of plasma cholinesterase. AB - OBJECTIVE: The goal of this study was to identify adverse effects of the atypical neuroleptic clozapine on liver function and lipid metabolism. METHODS: Data which included serum levels of clozapine and its hepatic metabolite N-desmethyl clozapine were collected from medical records of patients treated with clozapine and controls. RESULTS: We identified a clozapine-associated marked elevation of plasma cholinesterase (ChE) with unchanged levels of AST, ALT or g-GT. ChE was correlated to the serum level of clozapine and even closer to N-desmethyl clozapine. For the total patient group we observed significant correlations of ChE with the body-mass index and body weight. However, clozapine-treated patients and controls did not differ with regard to body-mass index, triglycerides, and cholesterol. CONCLUSION: We report for the first time a clozapine-associated and dose-dependent elevation of plasma ChE, which may be related to clozapine associated effects on hepatic lipid metabolism or ChE enzyme induction. PMID- 11881841 TI - Deficit states in schizophrenia and their association with the length of illness and gender. AB - The present study investigated the association between the frequency of deficit states (DS) and the length of illness and gender in schizophrenic patients. The following hypotheses were examined: 1) there is an association between the length of schizophrenic illness and the frequency of DS and 2) the frequency of DS in patients with comparable length of illness is higher in males than in females. Patients, included in the study, were consecutively hospitalized and diagnosed according to ICD-9 criteria (ICD-9: 295-295.9). Psychopathological assessment was performed in a standardized manner. Group differences were evaluated by using the t-test or the chi-square statistics. Variables with a possible impact on the occurrence of DS were entered into a backward multiple logistic regression model. 622 patients (total = 3914) were classified as having a DS. Logistic regression analysis revealed that the risk of having a DS was increased with a longer duration of illness (OR =1.68) and was reduced for female gender (OR = 0.56). The findings are in line with the hypothesis that apart from a neurodevelopmental origin, the schizophrenic illness may also have a progressive neurodegenerative component, which clinically emerges as DS. PMID- 11881842 TI - Inter-rater reliability of family history information on psychiatric disorders in relatives. AB - The family history method in psychiatric family studies is an important and necessary way of obtaining information on family members who are not available for personal interview. Studies on the validity of this method have shown that family history information on psychiatric disorders in relatives is neither accurate nor sensitive but highly specific. However, its inter-rater reliability has rarely been assessed, even though this is a prerequisite for adequate validity. In the present investigation we examined the inter-rater reliability of family history information obtained with a semi-structured and symptom-oriented interview. Forty informants were interviewed twice by two different raters within 3 and 20 days. The inter-rater reliability was found to be good for dementia (kappa=0.82, 95% CI=0.61-1.00), alcohol related disorders (kappa=0.93, 95% CI=0.80-1.00), for depressive disorders (kappa=0.72, 95% CI=0.42-1.00), anxiety disorders (kappa=0.75, 95% CI=0.41-1.00) and any psychiatric disorder (kappa=0.79, 95% CI=0.66-0.91). We concluded that the family history interview is a useful family study instrument that can be applied reliably by different raters for frequent psychiatric disorders. PMID- 11881844 TI - Dialysis--stands between deficit physiology and excess technology. PMID- 11881845 TI - Seven years experience with haemodialysis treatment for end stage renal disease in Dehradun. AB - Other than renal transplantation a maintenance programme with intermittent haemodialysis preserves life in patients of end stage renal disease. One hundred and eighty-three patients of end stage renal disease were taken for maintenance haemodialysis during last 7 years from April 1, 1991 to March 31, 1998 in a private set up in a north Indian city of Dehradun. There were 126 males (68.8%) and rest 57 (31.2%) were females. The youngest patient was 16 years old and oldest being 78 years old. Diabetic nephropathy in 55 (30.1%), bilateral contracted kidneys of unknown origin in 52 (28.4%), hypertensive nephropathy in 28 (15.3%), chronic glomerulonephropathy in 22 (12%), obstructive nephropathy in 11 (6%), tubulo-interstitial disease in 7 (3.8%), polycystic disease in 5 (2.7%) and post-transplant rejection in 3 (1.6%) contributed to the patients' pool. Only 35 patients (19.1%) had enough financial support to carry on clinically adequate dialysis with erythropoietin treatment on a regular basis. Average duration of haemodialysis therapy was only of 4 months. Only 3 patients were dialysed for more than 2 years and one patient is on haemodialysis for more than 5 years. Haemodialysis procedure perse was quite safe and accepted well with due care of technical considerations. Economic inadequacy was the major hurdle in providing more effective treatment and was also responsible for high fall outs. PMID- 11881843 TI - A follow-up study of 45 patients with elective mutism. AB - Forty five patients (23 boys and 22 girls) with elective mutism (8.7 +/- 3.6 years old), who were referred to a university department and a child guidance clinic within a 15-year-period, were followed up on average 12 years later. For 41 of them, sufficient information could be obtained at follow-up, and 31 patients could be investigated personally. At follow-up, an interview and a standardized psychopathological examination were carried out as well as two standardized biographic inventories. The main results were: 1) a high load of individual and family psychopathology was characteristic of the patients. The disorder started already at age 3 to 4 and referral age was 8 years on average. 2) In 16 out of 41 patients (39%), a complete remission could be observed. All other patients still revealed some communication problems. 3) The formerly mute patients described themselves as less independent, less motivated with regard to school achievement, less self-confident and less mature and healthy in comparison to a normal reference group. 4) A poor outcome could be best predicted by the variable "mutism within the core family"at the time of referral. PMID- 11881846 TI - Plasmcapheresis--principles and practice. AB - The concept of removal of blood "blood letting" was practised in ancient times. In the last four decades plasmapheresis, plasma exchange, or apheresis as the modality of treatment of certain specific disorders has become available. This article is a review of the principles of plasmapheresis. The equipment needed, the technique of plasmapheresis and guidelines for its use are discussed. PMID- 11881847 TI - Dialysis therapy in children. AB - Acute dialysis can be life saving for children suffering from acute emergencies due to acute renal failure, poisoning or inborn errors of metabolism causing severe organic acidaemia and hyperammonaemia. Chronic dialysis is life sustaining for children with chronic renal failure or end stage renal disease till renal transplantation is performed. The basic principles, indications, procedures, equipment, complications of acute and chronic dialysis in children are same as in adults. Dialysis can be performed in children at any age from newborn to adolescent period. In newborn, infants and preschool children (0-5 years age) haemodialysis is difficult due to technical problems associated with vascular access and haemodynamic instability. Peritoneal dialysis is simple, efficient, easy to perform, does not require highly sophisticated equipment or personnel and with low complication rate. For successful dialysis appropriate sizes of catheters, tubings, dialysers, small volume dialysate bags, etc, are required. These are now available in our country, although the cost of peritoneal dialysis fluid and catheters, etc, is 2-3 times higher than equipment for haemodialysis. Hence, continuous ambulatory peritoneal dialysis for chronic renal failure/end stage renal disease has not taken off yet in India. A team of experts including specially trained paediatric nephrologists, urologists, nurses, dieticians, technicians and social workers are needed to organise dialysis programme for children with end stage renal disease. Acute peritoneal dialysis should be made available in each paediatric department offering emergency services to children. PMID- 11881848 TI - Organ transplantation: present scenario and future strategies for transplant programme (specially cadaveric) in India: socio-administrative respects. AB - Organ transplantation was enacted by a law "Transplantation of Human Organ Act" in 1994 but still, getting the consent from the relatives of brain dead person is a very difficult task and hence cadaveric transplant accounts for a minimum number. In India, most of the transplantations carried out are related to living donor and very few are cadaveric. The poor status of cadaver transplantation may be attributed to the moral, emotional and religious beliefs and taboos that inhibit the relatives of the deceased to come forward to donate organ(s) of a brain dead person.Non-existence of a trained transplant co-ordinator who is the backbone of any successful transplant programme is another reason for poor response in cadaver transplantation. The great task is to motivate and prepare the relatives for organ donation of their near and dear ones. Transplant co ordinators are being prepared for motivating individuals or relatives for donation. To promote human organ transplantation government's initiative is very important. Mass media supported by the government can develop better awareness among the people. Non-government organisations (NGOs) can help in the similar ways. All hospitals are not authorised for the procedure of human organ transplantation. Other hospitals can help the process by informing the authorised hospitals about recent admission of potential donor (brainstem death). Role of transplant coordinator is crucial. He/she is the real inspiration to make agree the relatives for organ donation. Overall success of transplant programme is based on co-ordinated activity. Involvement of all agencies to motivate the person to pledge for organ donation during his/her life time is the first and the foremost requirement for successful planning and programme of organ transplantation. PMID- 11881849 TI - Dialytic support in acute renal failure. AB - Acute renal failure is the sudden failure on the part of the kidney to maintain normal biochemical homeostasis in the body. Medical causes of acute renal failure are fluid and electrolyte depletion, infectious diarrhoea, non-diarrhoeal infections, glomerulonephritis, poisoning with heavy metals, G-6-PD deficiency, snake bite and nephrotoxic drugs. Septic abortion is the most common cause of obstetric acute renal failure. Obstructive uropathy is an important cause of surgical acute renal failure. Guidelines to non-dialytic management of acute renal failure depend on the patient's condition which includes volume status, hyperkalaemia, acidaemia, uraemia and whether he/she is receiving medication and having adequate nutrition. Several dialytic methods are currently available for renal replacement therapy--intermittent haemodialysis and peritoneal dialysis are to name a few. Dialytic intervention in acute renal failure is usually considered when there is dinical evidence of uraemic symptoms or biochemical features of solute and fluid imbalance. Continuous renal replacement therapy implies to continuous replacement of renal function and it is limited to only a few big centres. PMID- 11881850 TI - Angio-access for haemodialysis--current perspective. AB - Vascular access for haemodialysis has seen many developments in recent times. Double lumen catheters introduced into wide bore veins have replaced the traditional Scribner shunt as temporary access thus reducing the complications and morbidity associated with it. Cuffed tunnelled internal jugular catheters and synthetic arteriovenous (AV) grafts usually made of polytetrafluoroethylene are the new additions to the vascular access armamentarium, but the AV fistula introduced in 1966 still remains the life-line for chronic haemodialysis patient. However, in elderly and diabetic patients, synthetic AV grafts are beneficial. The added advantage of synthetic grafts is short maturation time and multiple potential access sites. Access failure in 80% cases is due to venous stenosis and thrombotic episodes while infections or other complications are there in 20% cases. The complications of vascular access are not only a major cause of morbidity in haemodialysis patients but a major cost for the end stage renal disease programme. In western countries, access related morbidity accounts for almost 25% of all hospital stays for end stage renal disease patients and may contribute to as much as 50% of all hospitalisation cost. Access salvage includes prospective monitoring and treatment of outflow stenosis. The direct intra-access measure of blood flow by ultrasound dilution and duplex colour flow Doppler technique is the ideal method detecting venous outflow stenosis; however, conventional and digital subtraction angiography has an advantage, that total vasculature and blood flow may be visualised. The various treatment modalities for outflow stenosis include use of percutaneous transluminal angioplasty, stents and surgical correction. The lyses of secondary thrombosis can be done by surgical, medical and mechanical thrombolysis. The various methods being used to prevent graft stenosis include use of dipyridamole and radiation. The gene therapy holds promises for the future. PMID- 11881851 TI - Female infanticide and foeticide--the other view. PMID- 11881852 TI - Printing names of medicines on the strip. PMID- 11881853 TI - Unexplained infertility. PMID- 11881854 TI - Dermatoglyphics in 46, XY females. AB - Dermatoglyphics is known to be one of the best available diagnostic tools in genetic disorders. This paper aims to find out the diagnostic characteristic dermatoglyphic features in cytogenetically confirmed 46, XY female patients. The total number of patients studied (46, XY females) were 31 and the control consisted of 30 males and 30 females. Dermatoglyphic features, studied and tabulated, were: (a) Finger pattern frequency, (b) total finger ridge count (TFRC), absolute finger ridge count (AFRC), a-b ridge count, 'atd' angle and (c) palmar patterns eg, simian crease, Sydney line, hypothenar pattern, interdigital patterns. Results confirmed significant differences between the 46, XY females and the control groups: (i) 46, XY females had increased ulnar loops but decreased whorls as compared to control males (overall finger pattern frequency). (ii) The right thumb in 46, XY females had less whorls and more ulnar loops than both male and female control groups (individual finger pattern frequency). (iii) 46, XY females had lowered TFRC, AFRC, a-b ridge count than corresponding control groups (both control males and females). Significant differences were not observed for the 'atd' angle, interdigital patterns, hypothenar pattern, simian crease and Sydney line. PMID- 11881855 TI - Evaluation of fine needle aspiration and imprint cytology in the early diagnosis of breast lesions with histopathological correlation. AB - Fifty-one breast lumps were studied by fine needle aspiration cytology (FNAC) and scrape imprint cytology. Of the 51 cases, 28 were malignant and included infiltrating duct carcinoma (22), well differentiated adenocarcinoma (2), mucinous, papillary and lobular carcinoma (one each) and malignant lymphoma (one). Benign lesions were mastitis, fibroadenoma and fibrocystic disease. The diagnostic accuracy of FNAC was 90.2% and that of imprint 94.1% with no false negatives. Two imprint smears and one aspirate designated suspicious were proved to be benign. The combination of FNAC and imprint cytodiagnosis gave a diagnostic accuracy of 96% thus proving their value in the rapid diagnosis of breast lesions. PMID- 11881856 TI - Humoral immune responses in different clinical forms of tuberculosis. AB - To assess the antibody responses in different clinical forms of tuberculosis in an attempt to ascertain the specificity of such a response, so that it may be used as a diagnostic tool, 93 tuberculosis cases and 62 normal healthy controls were included in the study. All the cases were diagnosed as tuberculosis by clinical picture including radiological findings, blood picture, fine needle aspiration cytology (FNAC) or histopathological features, sputum examination or urine culture for acid-fast bacilli. IgM and IgG antibodies in the cases and controls were detected by ELISA using A60 antigen from M. bovis and compared with common diagnostic parameters of tuberculosis like presence of acid-fast bacilli in smear and tuberculin response. Antibody responses with A60 antigen has not been found to be specific and often failed to detect a new case. PMID- 11881857 TI - Non-interventional and interventional transvaginal ultrasound in present day infertility practice. AB - The diagnosis of cause and targeted management of infertility has emerged in leaps and bounds over the last few decades. Here the author has described the non interventional (eg, vascular study in ovulation, ultrasound study in polycystic ovarian disease and other pelvic pathological conditions) and interventional transvaginal ultrasound as evident in infertility practice today. She concluded with the few points as the possible future directions in infertility management. PMID- 11881858 TI - Management of endometriosis and its impact on infertility. AB - Endometriosis is defined as the presence of tissue, histologically similar to endometrium, in any other site but that of uterine cavity. Transplantation of endometrial tissue that comes as a retrograde menstrual flow is the most accepted possible theory. Endometriosis is associated with marked subfertility, but the mechanism of causation of infertility is poorly understood unless it has caused very definite anatomical distortion of internal genitalia. Management of infertility in endometriosis is discussed in detail. PMID- 11881859 TI - Preimplantation genetic diagnosis of chromosomal abnormalities by multicolour fluorescence in situ hybridisation. AB - Preimplantation genetic diagnosis (PGD) is an early diagnosis of genetic disorders, prior to the onset of pregnancy. PGD incorporates the latest techniques in assisted reproduction and molecular genetics. Embryos or oocytes are biopsied during culture in vitro and genetic analysis is carried out on the blastomeres or polar bodies. Embryos shown to be free of the genetic disease under investigation are transferred to the uterus. Multicolour fluorescence in situ hybridisation (FISH) is used to diagnose numerical and certain structural abnormalities of chromosomes in the embryo. The common probes used are for chromosomes 13, 18, 21, X and Y. FISH can also be used for PGD of translocations, when one of the parents is a carrier. PGD was carried out recently in 4 cases using multicolour FISH. In one of the embryos, trisomy 18 was detected. Tetraploidy was seen in another embryo. Only chromosomally normal embryos were transferred back to the uterus. Care has to be taken while interpreting FISH signals as the signal may be split, diffused, superimposed or in a different focus. PMID- 11881860 TI - Percutaneous epididymal sperm aspiration followed by intrauterine insemination--a new approach to achieve a pregnancy in infertility due to obstructive azoospermia. AB - Standard approach towards treatment of obstructive azoospermia whether congenital or acquired is mostly microsurgical epididymal sperm aspiration (MESA) and to some extent percutaneous epididymal sperm aspiration (PESA) both being followed by intracytoplasmic sperm injection (ICSI). However in the present series for sperm retrieval in case of obstructive azoospermia the author conducted a series of 55 cases of PESA along with only 4 cases of MESA. The epididymal sperm aspiration (ESA) was followed by intrauterine insemination (IUI) (whenever required quantity of motile sperm was collected) in place of ICSI with full consent of the couples. With the help of this ESA-IUI programme it was possible to achieve reasonable success rate viz, 20% pregnancy rate (PR) per cycle for PESA-IUI and 14.28% for MESA-IUI. PMID- 11881861 TI - Topiramate: a new safe and effective antiepileptic. AB - Twenty patients of either sex, with refractory partial epilepsy with or without secondary generalisation were entered in an open label study to evaluate the efficacy and safety of topiramate in them. Topiramate was used as an adjunctive therapy with an initial starting dose of 50 mg/day. The dose was then titrated upwards with increments of 50 mg per week, till a time the most effective and the best tolerated dose was reached. This most effective/tolerated dose was then continued for 6 months. Of the 17 patients entering the maintenance phase, 4 patients (24%) became seizure free, while a total of 14 patients (83%) out of 17 cases responded with a reduction in monthly seizures rate by 50% or more. Mean reduction of 68.9% was observed in monthly seizure rate during the maintenance phase. The median effective dose of topiramate was 600 mg per day. Five patients dropped out of the study due to adverse events such as anxiety, aggressiveness, rash, lethargy, etc. The central nervous system (CNS) related side effects such as dizziness, headache, and tremor were reported, which are commonly seen with other presently available antiepileptics like carbamazepine, phenytoin sodium, sodium valproate, etc, as well. Most adverse events, however, were mild and transient and did not interfere with the day to day activity of the patients. Topiramate was not associated with any abnormality in laboratory or neurological examination findings. The excellent response with topiramate therapy in Indian patients, uncontrolled with the available antiepileptics, as well as its good safety profile endorse the international efficacious and safe image of topiramate. PMID- 11881862 TI - Evaluation of efficacy and safety of nimesulide with betacyclodextrin vs nimesulide tablets in osteoarthritis. AB - Efficacy and safety of nimesulide as well as favourable tolerability have been tested in osteoarthritis in short term study and post-marketing survey. Here is a report which shows the superiority of nimesulide complexed with betacyclodextrin vs ninesulide tablet in osteoarthritis in a long term study. PMID- 11881863 TI - The sitting position in neurosurgery--not yet obsolete! PMID- 11881864 TI - Modulation of beta-adrenergic receptor subtype activities in perioperative medicine: mechanisms and sites of action. AB - This review focuses on the mechanisms and sites of action underlying beta adrenergic antagonism in perioperative medicine. A large body of knowledge has recently emerged from basic and clinical research concerning the mechanisms of the life-saving effects of beta-adrenergic antagonists (beta-AAs) in high-risk cardiac patients. This article re-emphasizes the mechanisms underlying beta adrenergic antagonism and also illuminates novel rationales behind the use of perioperative beta-AAs from a biological point of view. Particularly, it delineates new concepts of beta-adrenergic signal transduction emerging from transgenic animal models. The role of the different characteristics of various beta-AAs is discussed, and evidence will be presented for the selection of one specific agent over another on the basis of individual drug profiles in defined clinical situations. The salutary effects of beta-AAs on the cardiovascular system will be described at the cellular and molecular levels. Beta-AAs exhibit many effects beyond a reduction in heart rate, which are less known by perioperative physicians but equally desirable in the perioperative care of high risk cardiac patients. These include effects on core components of an anaesthetic regimen, such as analgesia, hypnosis, and memory function. Despite overwhelming evidence of benefit, beta-AAs are currently under-utilized in the perioperative period because of concerns of potential adverse effects and toxicity. The effects of acute administration of beta-AAs on cardiac function in the compromised patient and strategies to counteract potential adverse effects will be discussed in detail. This may help to overcome barriers to the initiation of perioperative treatment with beta-AAs in a larger number of high-risk cardiac patients undergoing surgery. PMID- 11881865 TI - The sitting position for neurosurgery in children: a review of 16 years' experience. AB - BACKGROUND: Use of the sitting position for neurosurgery is controversial. The main concern is the risk of venous air embolus (VAE) and its sequelae. METHODS: The paediatric neurosurgeons at our institution routinely use the sitting position for posterior fossa and pineal surgery, and a retrospective audit of the incidence of VAE from 1982 to 1998 has been performed. RESULTS: Venous air embolism, defined as a fall in end-tidal carbon dioxide pressure >0.4 kPa, was detected in 38 of 407 operations (9.3%). A fall in systolic arterial pressure >10% accompanied the VAE in nine out of 43 episodes (20.9%); this represents 2% of all operations. All VAE episodes responded promptly to treatment and there was no perioperative morbidity or mortality directly attributed to it. CONCLUSIONS: This is the largest study of the incidence of VAE in children undergoing neurosurgery. Our results suggest that the sitting position can be used safely for neurosurgery in children. PMID- 11881866 TI - Optimizing fluid therapy in mechanically ventilated patients after cardiac surgery by on-line monitoring of left ventricular stroke volume variations. Comparison with aortic systolic pressure variations. AB - BACKGROUND: Mechanical ventilation causes changes in left ventricular preload leading to distinct variations in left ventricular stroke volume and systolic arterial pressure. Retrospective off-line quantification of systolic arterial pressure variations (SPV) has been validated as a sensitive method of predicting left ventricular response to volume administration. We report the real-time measurement of left ventricular stroke volume variations (SVV) by continuous arterial pulse contour analysis and compare it with off-line measurements of SPV in patients after cardiac surgery. METHODS: SVV and SPV were determined before and after volume loading with colloids in 20 mechanically ventilated patients. RESULTS: SVV and SPV decreased significantly after volume loading and were correlated (r=0.89; P<0.001). Changes in SVV and changes in SPV as a result of volume loading were also significantly correlated (r=0.85; P<0.005). Changes in SVV correlated significantly with changes in stroke volume index (SVI) (r=0.67; P<0.005) as did changes in SPV (r=0.56; P<0.05). SVV determined before volume loading correlated significantly with changes in SVI (R=0.67; P <0.005). Using receiver operating characteristics curves, the area under the curve was statistically greater for SVV (0.824; 95% confidence interval: [CI] 0.64-1.0) and SPV (0.81; CI: 0.62-1.0) than for central venous pressure (0.451; CI: 0.17-0.74). CONCLUSIONS: Monitoring of SVV enables real-time prediction and monitoring of the left ventricular response to preload enhancement in patients after cardiac surgery and is helpful for guiding volume therapy. PMID- 11881867 TI - Reliability of epigastric auscultation to detect gastric insufflation. AB - BACKGROUND: We studied the reliability of epigastric auscultation to detect gastric insufflation in 30 anaesthetized, paralysed intubated patients. METHODS: A 16FG gastric tube was positioned with the tip in the mid-oesophagus with the proximal end attached to an injection port with a one-way valve. Four observers participated in the study. Observers were paired and each pair studied 15 patients. Each patient underwent four test sequences in random order, two by each observer. Each test sequence comprised one observer injecting different volumes of air (0.25 ml, 0.5 ml, 1 ml, 2 ml, 3 ml, 4 ml, 5 ml, 10 ml, 15 ml and 0 ml as a control) in random order whilst the second blinded observer listened with a stethoscope over the epigastrium. Each randomized volume was injected rapidly at 5 s intervals for 1 min. The number of injections required to detect air entering the stomach was recorded. The stomach was deflated between each test sequence. RESULTS: To detect air entering the stomach with 95% confidence, 11 injections were required for 0.25 ml; 7 for 0.5 ml; 3 for 1 ml; 2 for 2 ml and 3 ml, and I for > or =4 ml. The mean (range) inter- and intraobserver reliability was 0.73 (0.71-0.75) and 0.76 (0.76-0.89), respectively. The incidence of false positives was 21% (25/120) and the incidence of false negatives was 10% (103/1080), making the specificity and sensitivity 79% and 91%, respectively. CONCLUSIONS: We conclude that epigastric auscultation can detect gastric insufflation of 0.25 ml air after 11 breaths and > or = 4 ml air after one breath with 95% confidence. Inter- and intraobserver reliability is moderate to excellent. Epigastric auscultation should be repeated to reduce the risk of false positives. PMID- 11881868 TI - Deep topical fornix nerve block versus peribulbar block in one-step adjustable suture horizontal strabismus surgery. AB - BACKGROUND: We compared the efficacy of deep topical fornix nerve block anaesthesia (DTFNBA), which does not paralyse the extraocular muscles, with peribulbar block in patients undergoing one-step adjustable-suture horizontal strabismus surgery. Patients with a vertical, oblique squint were excluded from the study. METHODS: We studied 100 patients, allocated randomly to two groups. Group 1 (n=50) received peribulbar block with 5 ml of 1:1 mixture of 0.5% plain bupivacaine and 2% lignocaine supplemented with hyaluronidase 300 i.u. ml(-1). Group 2 (n=50) received DTFNBA with placement of a sponge soaked in 0.5% bupivacaine deep into the conjunctival fornices for 15 min. No sedation was given to either group. Analgesia was assessed by direct questioning of patients during the procedure. A three-point scoring system was used (no pain = 0, discomfort = 1, pain =2). If the pain score was 1, the patient was asked to look in the opposite direction to decrease the tension on the periosteal attachment of the muscle to relieve discomfort. If the pain score was 2 at any stage of the operation, general anaesthesia was given. RESULTS: In Group 2, significantly more patients (15) experienced discomfort than in Group 1 (no patients) (P<0.05), but general anaesthesia was not needed. CONCLUSIONS: DTFNBA is a useful technique for intraoperative adjustable-suture strabismus surgery. It does not alter muscle tone, thus allowing the surgeon to adjust the muscle sutures intraoperatively, and reducing the incidence of under- or over-correction of the squint in the immediate postoperative period. PMID- 11881869 TI - Cardiac arrest after administration of Omnipaque radiocontrast medium during endoluminal repair of abdominal aortic aneurysm. AB - We describe a case of acute cardiovascular collapse in a patient with end-stage renal failure undergoing endoluminal repair of an abdominal aortic aneurysm. The 61-yr-old man suffered cardiac arrest shortly after administration of radiocontrast medium (Omnipaque), during deployment of the endovascular device. He had received the same contrast solution for diagnostic angiography on the previous day. He was successfully resuscitated and recovered completely. The differential diagnosis and management are discussed. PMID- 11881870 TI - Skin necrosis following injection of non-steroidal anti-inflammatory drug. AB - We report two patients who developed skin necrosis following an intramuscular injection of non-steroidal anti-inflammatory drug into the anteriolateral area of their thigh. Both patients required further multiple operations and one developed life threatening septicaemia. They were left with disfiguring scars. Skin necrosis, although rare, is a recognized adverse reaction to intramuscular non steroidal anti-inflammatory injections. It is likely that the injections were, unwittingly, administered subcutaneously. It is important to ensure intra muscular administration, with an appropriate needle length. PMID- 11881871 TI - Recurrent neurological symptoms in a patient following repeat combined spinal and epidural anaesthesia. AB - A healthy woman developed neurological symptoms after two consecutive Caesarean sections under combined spinal and epidural anaesthesia. Amethocaine was used for spinal anaesthesia and mepivacaine for epidural anaesthesia on both occasions, and a combination of fentanyl and bupivacaine was continuously infused for pain relief after the second. Her symptoms on both occasions were similar, including pain in the buttocks of 7-11 days duration and numbness in the sacral area of 5-6 months. PMID- 11881872 TI - Loss of the guide wire: mishap or blunder? AB - We describe four cases of lost guide wires during central venous catheterization. Although percutaneous catheterization of central veins is a routine technique, it is a procedure requiring advanced operating skills, expert supervision, and attention to detail in order to prevent adverse effects. PMID- 11881873 TI - How to avoid fatal complications after central venous catheterization. PMID- 11881874 TI - How to avoid fatal complications after central venous catheterization. PMID- 11881876 TI - Is it safe to artificially ventilate paralysed patients through a laryngeal mask? PMID- 11881877 TI - Primary pulmonary hypertension in pregnancy. PMID- 11881878 TI - Remember the straight laryngoscope. PMID- 11881879 TI - Use of magnesium to treat tetanus. PMID- 11881880 TI - Myasthenia gravis unmasked by neuromuscular blockade. PMID- 11881881 TI - Potentially dangerous labeling of levobupivacaine. PMID- 11881882 TI - Oxygen and elective caesarean section. PMID- 11881883 TI - Fresh gas flow is not the only determinant of volatile agent consumption: a multi centre study of low-flow anaesthesia. AB - METHODS: Seven academic centres studied 302 patients, using desflurane, enflurane, halothane, or isoflurane using circle-systems and Drager Julian anaesthetic machines, with fresh gas flows (V(F)) of 3, 1, and 0.5 litre min(-1). Volatile agent partial pressures in the breathing system were recorded and agent consumptions measured by weighing. RESULTS: At these flows, desflurane consumption depended on V(F). In contrast, halothane consumption was not influenced by V(F). Isoflurane and enflurane showed differences in consumption between flows of 0.5 and 3 litre min(-1). Stepwise linear regression suggested that besides V(F), other factors influenced consumption of the more soluble agents (sex, age, weight, height, altitude, and temperature). The partial pressure ratios were independent of V(F) for desflurane (end-tidal to fresh gas=0.8), but the ratios of the more soluble agents varied with V(F) (end-tidal to fresh gas=0.3-0.7). CONCLUSIONS: At V(F) that involves significant re breathing, consumption of soluble agents depends only partially on V(F). These results can be explained using Mapleson's hydraulic analogue model. PMID- 11881885 TI - Blood transfusion in critical illness. PMID- 11881884 TI - Intrinsic positive end-expiratory pressure during one-lung ventilation of patients with pulmonary hyperinflation. Influence of low respiratory rate with unchanged minute volume. AB - BACKGROUND: We measured lung mechanics and gas exchange during one-lung ventilation (OLV) of patients with chronic obstructive pulmonary disease, using three respiratory rates (RR) and unchanged minute volume. METHODS: We studied 15 patients about to undergo lung surgery, during anaesthesia, and placed in the lateral position. Ventilation was with constant minute volume, inspiratory flow and FIO2. For periods of 15 min, RR of 5, 10, and 15 bpm were applied in a random sequence and recordings were made of lung mechanics and an arterial blood gas sample was taken. Data were analysed with the repeated measures ANOVA and paired t-test with Bonferroni correction. RESULTS: PaO2 changes were not significant. At the lowest RR, PaCO2 decreased (from 42 (SD 4) mm Hg at RR 15-41 (4) mm Hg at RR 10 and 39 (4) mm Hg at RR 5, P<0.01), and end-tidal carbon dioxide increased (from 33 (5) mm Hg at RR 15 to 35 (5) mm Hg at RR 10 and 36 (6) mm Hg at RR 5, P<0.01). Intrinsic positive end-expiratory pressure (PEEPi) was reduced even with larger tidal volumes (from 6 (4) cm H2O at RR 15-5 (4) cm H2O at RR 10, and 3 (3) cm H2O at RR 5, P<0.01), most probably caused by increased expiratory time at the lowest RR. CONCLUSION: A reduction in RR reduces PEEPi and hypercapnia during OLV in anaesthetized patients with chronic obstructive lung disease. PMID- 11881886 TI - Extreme value theory applied to postoperative breathing patterns. AB - BACKGROUND: There has been little published work on the statistical features of breath times in postoperative patients. We applied extreme value theory (a statistical method) to the variation in the timing of postoperative breathing. METHODS: We observed 49 patients 3-6 h after a variety of surgical procedures, once they had achieved a stable breathing pattern. The breathing patterns could be one of the three types predicted by the extreme value model. 'Finite' breathing patterns (n=30) have a finite upper limit of duration for any apnoea. Patients that displayed one of the other two patterns ('standard' and 'extended') have, potentially, no limit in duration of apnoea. RESULTS: The type of breathing pattern observed in each patient was not reliably identified by most of the commonly used risk factors (age, type of surgery, opioid type, dose, and route of administration). A finite pattern was observed in 13 of 26 patients receiving epidural (vs 17 of 23 parenteral analgesia: P=0.15), and 15 of 19 receiving morphine (vs 15 of 30 other opioids: P=0.05). The patients with 'finite' patterns were also significantly less drowsy (score 1.04 (0.92) vs 1.62 (0.62), P<0.05). CONCLUSIONS: The breathing pattern was not related to mean breath times, suggesting that the prevalence of apnoeas cannot be reliably predicted by measurement of the respiratory rate alone. PMID- 11881887 TI - Randomized controlled trial to investigate influence of the fluid challenge on duration of hospital stay and perioperative morbidity in patients with hip fractures. AB - BACKGROUND: A prospective, randomized controlled trial comparing conventional intraoperative fluid management with two differing methods of invasive haemodynamic monitoring to optimize intraoperative fluid therapy, in patients undergoing proximal femoral fracture repair under general anaesthesia. METHODS: Ninety patients randomized to three groups; conventional intraoperative fluid management (Gp CON, n=29), and two groups receiving additional repeated colloid fluid challenges guided by central venous pressure (Gp CVP, n=31) or oesophageal Doppler ultrasonography (Gp DOP, n=30). Primary outcome measures were time to medical fitness to discharge, hospital stay and postoperative morbidity. RESULTS: The fluid challenge resulted in significantly greater perioperative changes in central venous pressure between Gp CVP and Gp CON (mean 5 (95% confidence interval 3-7) mm Hg) (P<0.0001). Important perioperative changes were also shown in Gp DOP with increases of 49.4 ms (19.7-79.1 ms) in the corrected flow time, 13.5 ml (7.4-19.6 ml) in stroke volume, and 0.9 (0.49-1.39) litre min(-1) in cardiac output. As a result, fewer patients in Gp CVP and Gp DOP experienced severe intraoperative hypotension (Gp CON 28% (8/29), Gp CVP 9% (3/31), Gp DOP 7% (2/30), P=0.048 (chi-squared, 2 degrees of freedom (df). No differences were seen between the three groups when major morbidity and mortality were combined, P=0.24 (chi-squared, 2 df). Postoperative recovery for survivors, as defined by time to be deemed medically fit for discharge, was significantly faster, in comparison with Gp CON, in both the Gp CVP (10 vs 14 (95% confidence interval 8-12 vs 12-17) days, P=0.008 (t-test)), and Gp DOP (8 vs 14 (95% confidence interval 6-12 vs 12 17) days, P=0.023 (t-test). There were no significant differences between groups, for survivors, with respect to acute orthopaedic hospital and total hospital stay. CONCLUSIONS: Invasive intraoperative haemodynamic monitoring with fluid challenges during repair of femoral fracture under general anaesthetic shortens time to being medically fit for discharge. PMID- 11881888 TI - Double-blind, placebo-controlled analgesic study of ibuprofen or rofecoxib in combination with paracetamol for tonsillectomy in children. AB - BACKGROUND: The analgesics used for paediatric tonsillectomy may be associated with side-effects such as sedation, respiratory depression and vomiting (opioids) or increased bleeding [non-steroidal anti-inflammatory drugs (NSAIDs)]. In our institution, we employ a combination of paracetamol, NSAID and opioid, although there is no published evidence of analgesic benefit from adding NSAIDs to paracetamol in children. METHODS: This randomized, double-blinded clinical study examined the analgesic effectiveness of combining paracetamol (20 mg kg(-1)) with rofecoxib (0.625 mg kg(-1)), ibuprofen (5 mg kg(-1)) or placebo as premedication for (adeno)tonsillectomy (n=98) in children aged 3-15 yr. Intravenous fentanyl 1 2 microg kg(-1) was given intraoperatively. Regular oral paracetamol (15 mg kg( 1), 4 hourly) was given after operation and could be supplemented on request from the child with oral ibuprofen 5 mg kg(-1) or oral codeine 1 mg kg(-1). The primary outcome variable was need for early supplementary analgesia (within 2 h after surgery). RESULTS: The addition of ibuprofen to paracetamol reduced the need for early analgesia from 72% to 38% of children (difference 34%; 95% confidence interval 4-64%). The addition of rofecoxib to paracetamol did not significantly alter the need for early analgesia (68 vs 72%). Pain scores were higher in those children who required early analgesia. There were no differences between the groups in operative blood loss or complications, total 24-h analgesic consumption, pain scores at 4 and 8 h, vomiting or antiemetic use. CONCLUSION: This study provides evidence to support the combination of ibuprofen (but not rofecoxib) with paracetamol for perioperative analgesia in children. PMID- 11881889 TI - Effects of intracoronary calcium chloride on regional oxygen balance and mechanical function in normal and stunned myocardium in dogs. AB - BACKGROUND: Brief myocardial ischaemia has been demonstrated to result in mechanical and coronary endothelial dysfunction, in which calcium may play a role. We examined whether the mechanical and vascular responses to calcium are altered in postischaemic, reperfused myocardium. METHODS: Regional myocardial oxygen consumption (MVO2), mechanical function and coronary blood flow (CBF) in response to calcium chloride (0.10, 0.25, 0.50 and 0.75 mg ml(-1) of CBF) directly infused into the left anterior descending (LAD) artery were determined before (normal) and 30 min after a 15-min-period of LAD occlusion (stunned) in an open-chest canine model. Percentage segment shortening (%SS) and percentage postsystolic shortening (%PSS) in the LAD territory were determined using ultrasonic crystals and CBF using a Doppler transducer. Myocardial extraction of oxygen (EO2) and lactate (Elac) was calculated. RESULTS: The infusion of calcium chloride resulted in dose-dependent increases in %SS and MVO2 but did not affect %PSS in normal myocardium. These changes were accompanied by parallel increases in CBF, resulting in no change in EO2. In stunned myocardium, the responses to calcium chloride were not significantly altered, with the exception of a reduction in %PSS. However, ischaemia and reperfusion itself significantly reduced %SS and Elac and increased %PSS. CONCLUSIONS: These data suggest that calcium chloride improves regional systolic and diastolic function both in normal and stunned myocardium. Calcium chloride is unlikely to cause direct coronary vasoconstriction or to deteriorate regional mechanical function in postischaemic myocardium. PMID- 11881890 TI - Airway management in the emergency department. PMID- 11881891 TI - Concurrent ketamine and alfentanil administration: pharmacokinetic considerations. AB - BACKGROUND: A ketamine-alfentanil combination has been suggested for total i.v. anaesthesia. We determined the pharmacokinetics of ketamine and alfentanil, alone and together, in three groups of adult male rats, to assess any pharmacokinetic interaction. METHODS: Group 1 animals were infused with i.v. ketamine for 5 min; in group 2, constant low plasma concentrations of alfentanil were maintained by computer-controlled infusion; in group 3, the treatments were combined. Serial plasma and terminal tissue concentrations were measured by high performance liquid chromatography or gas chromatography-mass spectrometry. RESULTS: In the presence of alfentanil, the mean plasma ketamine concentration-time area under the curve (AUC) value was significantly lower (by 13%, P<0.05), while clearance (CIT) and volume of distribution (Vss) were significantly higher (by 16 and 28%, respectively, both P<0.05). Tissue:plasma distribution coefficients for ketamine in the presence of alfentanil were significantly higher in forebrain (by 128%, P<0.005), hindbrain (by 207%, P<0.01), gut (by 254%, P<0.005), and fat (by 344%, P<0.0001). Mean AUC values for alfentanil did not differ significantly in the presence of ketamine, but alfentanil tissue concentrations were significantly lower in forebrain (by 77%, P<0.0001), hindbrain (by 28%, P<0.01), heart (by 33%, P<0.01), lung (30%, P<0.05), and gut (by 21%, P<0.05). Corresponding tissue:plasma distribution coefficients were significantly lower for forebrain (by 69%, P<0.0001) alone. CONCLUSIONS: The finding that the distribution of ketamine into the brain was increased by low plasma concentrations of alfentanil could have important clinical applications for pain management. PMID- 11881892 TI - Genomic analyses of bacterial respiratory and cytochrome c assembly systems: Bordetella as a model for the system II cytochrome c biogenesis pathway. AB - An analysis of thirty-three genomes of selected bacteria for the presence of specific respiratory pathways and cytochrome c biogenesis systems has led to observations on respiration and biogenesis. A table summarizing these results is presented. The data suggested that Bordetella pertussis would be an excellent genetic model to study the System II cytochrome c biogenesis pathway. These observations are discussed and the results of genetic studies on System II biogenesis in B. pertussis are presented as a case for the power of comparative genomics. System II is present in organisms as diverse as Helicobacter, Neisseria, Porphyromonas, mycobacteria, cyanobacteria, and plants (chloroplasts), indicating this pathway's prominence and that horizontal transfer of system II (and/or System I) must have occurred on multiple occasions. PMID- 11881893 TI - Bacteriophage therapy of infectious diseases in aquaculture. AB - Bacteriophages may be candidates as therapeutic agents in bacterial infections. Here we describe the protective effects of phages against experimentally induced bacterial infections of cultured fish and discuss the potential for phage therapy in aquaculture. PMID- 11881894 TI - The ubiquitous ThrE family of putative transmembrane amino acid efflux transporters. AB - We here report sequence analyses of a newly described family of putative amino acid exporters, the ThrE family. Homologues were identified in select bacteria, archaea and eukaryotes, but only in the fungal kingdom of eukaryotes. These proteins can exist either as single polypeptide chains or as pairs of polypeptide chains. Computational evidence suggests that these proteins exhibit 10 transmembrane alpha-helical segments (TMSs), having arisen from a five TMS precursor by an early intragenic duplication event. The phylogenetic tree of the ThrE family reveals that most proteins cluster according to organismal phylogeny with only a few exceptions, suggesting that the former proteins are orthologues. All family members exhibit hydrophilic N-terminal (and occasional C-terminal) extensions that show limited sequence similarity with a domain of unknown function found in many peptidases and proteases. The significance of these observations is discussed. PMID- 11881895 TI - Molecular cloning and functional characterisation of VanX, a D-alanyl-D-alanine dipeptidase from Streptomyces coelicolor A3(2). AB - The vanX gene which encodes a D-alanyl-D-alanine dipeptidase is critical for vancomycin resistance in enterococci. A putative vanX gene from Streptomyces coelicolor A3(2), which is not known for vancomycin production, was identified by homology-based analysis and cloned by polymerase chain reaction. The S. coelicolor vanX gene was heterologously expressed in Escherichia coli BL21(DE3) and enzymatic assays of soluble protein fractions of VanX revealed a 93-fold increase in dipeptidase activity as compared to the nonrecombinant control, thus confirming its functionality. Interestingly, S. coelicolor was also found to be of intermediate resistance to vancomycin although it does not produce vancomycin, thus suggesting the role of VanX in defence or immunity. As such, the prevalence of vanX genes in the environment may be more common than previously thought. PMID- 11881896 TI - Molecular characterization of a Spanish isolate of Mycobacterium malmoense. AB - We present the genomic characterization of a Spanish isolate of Mycobacterium malmoense. Identification and molecular characterization of the microorganism were performed using the technique of PCR-restriction fragment length polymorphism analysis (PRA) and sequencing of the 16S and the ITS spacer (16S-23S rDNA intergenic spacer region). It was seen that our isolate is very similar to the strain described in Sweden, but differs in the sequencing of the ITS from the isolate in Great Britain. Molecular techniques facilitate a rapid diagnosis and enable a better understanding of the epidemiology of this microorganism. PMID- 11881897 TI - Differentiation of leptospires of the serogroup Pomona by monoclonal antibodies, pulsed-field gel electrophoresis and arbitrarily primed polymerase chain reaction. AB - All reference strains described as representing separate serovars belonging to the serogroup Pomona and a clinical leptospiral isolate (LP2) from this serogroup were analyzed using a battery of 9 monoclonal antibodies, pulsed-field gel electrophoresis (PFGE) and arbitrarily primed polymerase chain reaction (AP-PCR). Monoclonal antibody analysis provided taxonomic results which were in agreement with the current classification of the serogroup Pomona into six serovars and allowed the classification of the isolate LP2 in the serovar pomona. PFGE and AP PCR, although in general agreement with monoclonal antibody analysis, also were able to demonstrate some differences in the restriction patterns of strains Pomona, Monjakov and CB. These results indicate that these strains, grouped within serovar pomona after the introduction of bacterial restriction endonuclease analysis as the typing method, but formerly described as representing separate serovars (pomona, monjakov and cornelli, respectively), are similar but not identical to one another. This was also the case with strains 5621, the serovar mozdok reference strain, and K1, formerly described as serovar dania reference strain, but currently recognized to be a mozdok-like strain. These findings suggest that the deletion of some serovars within the serogroup Pomona, namely mozdok, cornelli, and dania, should be reconsidered. Thus, PFGE appears to be a useful tool for the serovar identification of leptospires belonging to the serogroup Pomona and for shedding light on the problem of their classification. PMID- 11881898 TI - Bacteriocin production by Actinobacillus actinomycetemcomitans isolated from the oral cavity of humans with periodontal disease, periodontally healthy subjects and marmosets. AB - The ability of Actinobacillus actinomycetemcomitans to produce bacteriocin has rarely been reported. Antagonistic substance production may confer an important ecological advantage for the producer microorganisms, especially in a competitive ecosystem such as the oral cavity. In the present study, 75 A. actinomycetemcomitans strains isolated from the oral cavity of human patients with periodontal disease, periodontally healthy subjects and marmosets, as well as two reference strains (A. actinomycetemcomitans ATCC 29523 and FDC Y4) were evaluated for auto-, iso-, and heteroantagonistic activity. Fifty-one (68.00%) strains exhibited antagonistic activity; heteroantagonism was observed more often than isoantagonism. Isolated strains antagonized 17 different species of gram positive and gram-negative bacteria from the oral and nonoral microbiota. Sensitivity to heat and to proteolytic enzymes constituted strong evidence that the antagonistic substance has a proteic nature. Taken together, our data enabled us to confirm that the antagonistic substance detected was a bacteriocin. The wide spectrum of activity indicates the possibility that more than one antagonistic substance is produced and that these substances play an important role in the ecological balance of the oral ecosystem. PMID- 11881899 TI - Crosses between Saccharomyces cerevisiae and Saccharomyces bayanus generate fertile hybrids. AB - Crossings between strains of Saccharomyces cerevisiae and Saccharomyces bayanus were carried out. Genetic, molecular and electrophoretic karyotyping data indicated that interspecific hybrids were obtained. The hybrid cells segregated "grande" and "petite" colonies, and the latter ranged between 20 and 50%; unlike "grande" colonies, "petite" colonies did not sporulate and did not ferment maltose. In the hybrids, the extent of sporulation varied between 10 and 20%; only very rare asci (around 10(-4)) held viable ascospores. Clones from the viable ascospores sporulated and produced asci with viable ascospores able to give mating with spores from both hybrid derivatives and parental species. Fertile asci could derive from allotetraploid cells generated by endomitotic events in allodiploid cells, a mechanism that enables overcoming the species barrier between S. cerevisiae and S. bayanus. PMID- 11881900 TI - Life and death of dried prokaryotes. AB - The removal of water through air drying damages membranes, proteins and nucleic acids and is lethal to the majority of organisms. Nevertheless, some vegetative cells of bacteria and cyanobacteria survive extreme desiccation. Understanding the mechanisms of their desiccation tolerance is an important issue in cell biology and holds promise for the metabolic engineering of desiccation-sensitive cells. PMID- 11881901 TI - Effect of inversion on the recognition of external and internal facial features. AB - The purpose of the present study was to find out whether inversion affects recognition of external and internal facial features. 24 participants matched, under two experimental conditions (pair and multiple-choice matchings), upright target faces with three categories of facial test stimuli: full faces, external features and internal features, which were presented in either upright or inverted orientations. Data analysis showed that matching of facial stimuli was faster, more accurate and more consistent under upright than under inverted orientations for all stimulus categories; mostly for full faces, and least for internal features. As a rule, there were no speed-accuracy trade-offs. Implications of the data for accounts of the inversion effect in face recognition in terms of a shift from configurational to componential processing were discussed. PMID- 11881902 TI - Time estimation: does the reference memory mediate the effect of knowledge of results? AB - This study presents evidence for the role of knowledge of results (KR) in the estimation of medium time intervals (4-12 s durations). Experiment 1 tested the hypothesis that KR operates at the same stage of temporal processing as attention. The absence of an interaction of attention and KR is taken as evidence against this hypothesis. Experiments 2 and 3 further studied the effects of KR in a reproduction and a production task, respectively. The effects of KR were small in the reproduction and substantial in the production task. The results are interpreted in terms of a clock-based timing model and the findings taken together converge on the interpretation that KR affects the reference memory (RM), rather than the other components of the model. PMID- 11881903 TI - False memories and lexical decision: even twelve primes do not cause long-term semantic priming. AB - Semantic priming effects are usually obtained only if the prime is presented shortly before the target stimulus. Recent evidence obtained with the so-called false memory paradigm suggests, however, that in both explicit and implicit memory tasks semantic relations between words can result in long-lasting effects when multiple 'primes' are presented. The aim of the present study was to investigate whether these effects would generalize to lexical decision. In four experiments we showed that even as many as 12 primes do not cause long-term semantic priming. In all experiments, however, a repetition priming effect was obtained. The present results are consistent with a number of other results showing that semantic information plays a minimal role in long-term priming in visual word recognition. PMID- 11881904 TI - Spatial intention-response compatibility. AB - In the current investigation the assumption that intentions and actions are represented in the same medium or code (common coding hypothesis of intention and action) was tested and confirmed [Q. J. Exp. Psychol. 52 (1999) 1; Eur. J. Cognit. Psychol. 9 (1997) 129]. In two experiments it is shown that the intention to produce a spatial consequence in the environment to an otherwise spatially neutral stimulus leads to a shorter response, if the responses and the intended consequences share relative positions compared to a situation where this is not the case. In two control experiments it is ruled out that this spatial intention response compatibility effect is due to an intention-independent expectancy of the consequences of the correct responses. PMID- 11881905 TI - Specialist musical training and the octave illusion: analytical listening and veridical perception by pipe organists. AB - The octave illusion is a useful tool for investigation of the contribution of specialist training to auditory perception. The stimulus that induces the illusion involves two tones with a frequency ratio of 2:1, presented dichotically, and with ear of presentation reversed every 250 ms. Most listeners report hearing a single tone that alternates from high in the right ear to low in the left ear [Scientific American 233 (1975) 92-104]. The first experiment investigated the hypothesis that musical training contributes to veridical perception of an ambiguous stimulus. As hypothesized, participants with the highest level of musical training were more likely to perceive the stimulus veridically. Exploring the effects of specialist training, Experiment 2 contrasted expert pipe organists with other instrumentalists. As hypothesized, participants expert in playing pipe organ--an instrument with harmonic and spatial features similar to those of the octave illusion--were more likely to perceive the stimulus veridically. The results have implications for plasticity of the auditory system and the analytical listening that accompanies specialist, intensive training and rehearsal. PMID- 11881906 TI - Mediators of age-related differences in recollective experience in recognition memory. AB - This study examined states of awareness with the Remember/Know paradigm during verbal recognition memory in young and old adults. Following the presentation of a word list, subjects undertook a recognition test and indicated whether they could consciously recollect its prior occurrence (R) or recognize it on some other basis, without conscious recollection (K). In this individual-difference approach we also incorporated various processing-speed and working-memory measures to study the link between aging, states of awareness and processing resources. The results revealed that, compared to younger adults, older adults exhibited a decline in the amount of R responses during the recognition test whereas the amount of K responses did not change. Structural equation modeling indicated that a slower processing speed associated with a limited working-memory capacity is a key to explaining age-related variance in conscious recollection. The findings offer further support for the distinction between remembering and knowing and for the processing-resources hypothesis of aging. PMID- 11881907 TI - Timing of finger tapping to frequency modulated acoustic stimuli. AB - This study examined the timing of synchronous finger tapping to continuous frequency modulation (FM) and to click trains. Tapping to click trains was found to statistically significantly anticipate the acoustic stimulus. Tapping to continuous FM occurred before the instantaneous frequency rose through its mean value (i.e. at zero phase of the sinusoidal FM). The anticipation of zero phase of the FM was similar in magnitude to the anticipation of the click stimuli. However, there was a systematic departure from this timing when the FM depth was varied, the cause of which is unclear. The perceived timing of acoustic stimuli will influence the timing of motor responses to the stimuli. These results may therefore be relevant to the timing of perceptual centres of acoustic stimuli including speech. PMID- 11881908 TI - Breast and ovarian cancer screening practices in healthy women with a strong family history of breast or ovarian cancer. AB - Studies in women with a family history of cancer demonstrate a wide variability in the uptake of cancer screening measures. Little data exist regarding the breast and ovarian cancer screening practices of women who are members of hereditary breast cancer families. In order to address this issue, we examined the screening behaviors and the determinants of screening in a clinic based group of 216 women with a strong family history of breast or ovarian cancer who were participating in a free genetic counseling and testing research program. At baseline, prior to obtaining genetic counseling or testing, 50% of women ages 30 39, 83% of those age 40-49, 69% of those 50-64, and 53% of those >65 reported having a mammogram in the prior year. Adherence to mammography recommendations was correlated with age, number of relatives with breast cancer, and income. Twenty percent of participants had at least one CA- 125 performed and 31 % had ever obtained a screening ultrasound. Having at least one relative with ovarian cancer was very strongly associated with ovarian cancer screening [OR = 12.3, 95% CI = 4.6-33 for CA-125; OR=4.9, 95% CI=2.4, 10.1 for ultrasound]. No association between cancer worries/distress and either breast or ovarian cancer screening was found. In conclusion, the breast and ovarian screening uptake in healthy women from hereditary breast cancer families is suboptimal, even for women over age 50, for whom annual mammography is clearly indicated. These findings indicate a need for better education about screening guidelines for high-risk women. PMID- 11881909 TI - Self-reported dietary habits, overall dietary quality and symptomatology of breast cancer survivors: a cross-sectional examination. AB - Little information is available about the relationship between quality of life of women who have survived breast cancer (specifically, symptoms including those of menopause and depression) and the quality of their diet. In this cross-sectional study, 117 women with known primary breast cancer completed a self-administered food frequency questionnaire (FFQ) reflecting usual diet during the past year, a Survey of Feelings and Attitudes using the Center for Epidemiologic Studies Depression scale (CES-D) and a survey that includes menopausal symptoms among others common to women with a history of breast cancer. When women's responses to the FFQ were scored using the Healthy Eating Index (HEI), most often diets were evaluated as those that 'need improvement' with a mean total HEI score of 67.2. With regard to the CES-D scores, study women averaged 9.5, with 19 women being classified as clinically depressed. HEI and CES-D scores were inversely related (p = -0.22, p = 0.02). A negative correlation was also observed between energy adjusted calcium intakes and CES-D scores (p = -0.19, p = 0.04). Clinical depressed women had not only lower HEI scores and calcium intakes, but also lower grain and variety scores. Comparisons to national data for disease-free women and that available for those with breast cancer suggest that our study women consumed diets low in energy and dietary variety. Diet quality may be an important factor influencing the manifestation of depressive symptoms in breast cancer survivors or conversely, poorer diet quality may be an outcome of depression. PMID- 11881910 TI - Tamoxifen-induced increases in cytoplasmic free Ca2+ levels in human breast cancer cells. AB - Tamoxifen has been shown to increase cytoplasmic free Ca2+ levels [Ca2+]i in renal tubular cells and bladder cancer cells, and to after Ca2+ signaling in MCF 7 breast cancer cells. The present study examined the effect of tamoxifen on [Ca2+], in ZR-75-1 human breast cancer cells using fura-2 as an indicator. Tamoxifen increased [Ca2+]i at a concentration above 2 microM with an EC50 of 5 microM. Removing extracellular Ca2+ reduced the response by 48+/-2%. In Ca2+-free medium, after tamoxifen-induced [Ca2+]i increased had returned to baseline, adding 3 mM Ca2+ increased [Ca2+]i in a concentration-dependent manner. Further, pretreatment with 10 microM tamoxifen abolished the [Ca2+]i increase induced by 1 microM thapsigargin (an endoplasmic reticulum Ca2+ pump inhibitor); and conversely, pretreatment with thapsigargin prevented tamoxifen from releasing more Ca2+. Tamoxifen (10 microM)-induced Ca2+ release was not changed by inhibiting phospholipase C activity with 2 microM U73122. Trypan blue exclusion assay revealed that tamoxifen (1-10 microM) did not alter viability after 1 min of incubation, but killed 10% of cells after 3-10 min of incubation. Together, this study shows that tamoxifen (>2 microM) induced a significant, immediate increase in [Ca2+]i in ZR-75-1 breast cancer cells. Tamoxifen acted by releasing Ca2+ from the endoplasmic reticulum Ca2+ stores in a manner independent of phospholipase C activity, and by inducing Ca2+ entry from extracellular medium. Tamoxifen may be of mild cytotoxicity after acute exposure. PMID- 11881911 TI - In vitro cytotoxicity of bismuth-213 (213Bi)-labeled-plasminogen activator inhibitor type 2 (alpha-PAI-2) on human breast cancer cells. AB - Metastasis is the principal cause of death in breast cancer patients. New and improved treatments for eradicating micrometastases are needed. To this end, a novel alpha-emitting protein construct, 213Bi-labelled plasminogen activator inhibitor type-2 (PAI-2) (alpha-PAI-2), was evaluated in vitro. This construct exploits: (a) the overexpression of the cell-surface receptor bound urokinase plasminogen activator (uPA) in the metastatic spread of breast cancer cells; (b) the binding and inhibition of receptor-bound uPA by PAI-2; and (c) the high cytotoxicity of alpha radiation. High labeling efficiencies and stability of 213Bi bound to human recombinant PAI-2 conjugated with cyclic diethylenetriaminepentaacetic acid anhydride were achieved (greater than 90%). The uPA inhibitory activity of the chelated PAI-2 was maintained as determined by complex formation with uPA and by inhibition of uPA activity. Furthermore, the reactivity of alpha-PAI-2 was confirmed in a cell assay as this construct was highly cytotoxic to breast cancer cell lines that express active, receptor bound uPA. The specificity of alpha-PAI-2 targeting was shown using several controls. Firstly, an active uPA blocking agent that limits PAI-2 binding significantly improved cell survival by a factor greater than three. Secondly, a non-specific alpha-BSA construct had minimal cytotoxic effect. Moreover, alpha-PAI-2 was not cytotoxic to freshly isolated normal human leukocytes, confirming that cells which do not contain active, receptor bound uPA cannot be targeted by alpha-PAI 2. In conclusion, we have validated, in vitro, the potential of alpha-PAI-2 as a novel therapeutic agent for breast cancer. PMID- 11881913 TI - Selective axillary surgery in breast cancer patients based on positron emission tomography with 18F-fluoro-2-deoxy-D-glucose: not yet! AB - We prospectively evaluated 31 patients with invasive breast cancer. Preoperative positron emission tomography (PET) with 18F-fluoro-2-deoxy-D-glucose (18F-FDG) for detection of axillary lymph node metastases was compared with the histopathologic status of the sentinel lymph node (SLN). Sensitivity of PET imaging was 43%, specificity and negative predictive value were 94 and 67%, respectively. The smallest metastasis detected by PET measured 3 mm in diameter. The results of this study suggest that detection of small axillary lymph node metastases is limited by the currently achievable spatial resolution of PET imaging. Selective axillary surgery in breast cancer patients based on 18F-FDG PET is yet not possible. PMID- 11881912 TI - Prognostic significance of tumor cell proliferation analyzed in fine needle aspirates from primary breast cancer. AB - The objective of this study was to analyze the role of proliferating fraction (PF) measured with Ki-67/MIB-1 antibody in a large series of preoperative fine needle aspirate (FNA) biopsies as a prognosticator of disease recurrence. The study comprised 732 patients who all had a conclusive cytological diagnosis of breast cancer. The follow-up time ranged from 1.2 to 10.2 years with a median of 5.7 years. In multivariate analysis Ki-67/MIB-1 value was a strong (p < 0.001) significant, prognosticator of disease recurrence free interval (DRFI) independent of lymph node status, progesterone receptor content, and tumor size. In the subgroup analysis of 430 node-negative patients the distant recurrence free rate after 5 years was 94.4% in patients with Ki-67/MIB-1 value < 15% compared to 88.7% in patients with Ki-67/MIB-1 value > or = 15% (p = 0.03). Test of the interaction between tumor size and the value of PF revealed a p-value of 0.06. If the patients, in addition, had a tumor size >20 mm the distant recurrence-free rate after 5 years was 93.2% if Ki-67/MIB-1 < 15% compared to 80.7% in patients with Ki-67/MIB-1 value > or = 15%. This difference was statistically significant (p < 0.01). For patients with tumors <20mm Ki-67/MIB-1 value did not add any prognostic information. In the subgroup of 302 node positive patients the distant recurrence-free rate after 5 years was 86.0% in patients with Ki-67/MIB-1 value < 15% compared to 70.6% in patients with Ki 67/MIB-1 value > or = 15% (p < 0.01). We conclude that PF assessed by Ki-67/MIB-1 antibodies in preoperative FNA biopsies has a significant prognostic value independent of lymph node status, PgR status and tumor size. To our knowledge, this is the first study demonstrating PF, which can contribute prognostic information when analyzed in preoperative smears. PMID- 11881914 TI - Factors associated with success of the extreme drug resistance assay in primary breast cancer specimens. AB - The extreme drug resistance (EDR) assay has not been widely studied in the setting of non-metastatic breast cancer. We evaluated the feasibility of performing the assay in 144 primary breast tumor specimens from two institutions by determining the rate of successful tumor culture for assays, number of drugs evaluated per assay, and time from tumor biopsy to receipt of results. We also sought to determine factors that are associated with assay success. An exploratory analysis was performed to detect possible associations between estrogen receptor (ER), progesterone receptor (PR) and HER2/NEU over-expression and extreme drug resistance demonstrated by the assay for specific chemotherapeutic agents. Of 144 tumor specimens submitted, tumor was successfully cultured for assay in 101(70%) of cases. A median of five drugs was evaluated per assay (range 2-9). Results were obtained in a median of 8 days (range 2-29). Young age, high tumor grade, PR negativity, and higher tumor submission weight were predictive for a successful assay. EDR was observed in 7-15% of tumors to doxorubicin, cyclophosphamide, 5-fluorouracil (5FU) and mitoxantrone, but EDR to paclitaxel was observed in 35%. Extreme drug resistance to 5-FU was associated with negative ER and PR status. There was a trend toward association between EDR to paclitaxel and HER2/NEU over-expression. The EDR assay may be successfully performed in the majority of tumors, and assay results are available in a timely fashion such that adjuvant treatment drug selection could be guided by results. These results may be helpful for designing possible future trials that evaluate the assay's role in adjuvant chemotherapy selection. PMID- 11881915 TI - Speech perception and spoken word recognition: past and present. AB - OBJECTIVE: The scientific study of the perception of spoken language has been an exciting, prolific, and productive area of research for more than 50 yr. We have learned much about infants' and adults' remarkable capacities for perceiving and understanding the sounds of their language, as evidenced by our increasingly sophisticated theories of acquisition, process, and representation. We present a selective, but we hope, representative review of the past half century of research on speech perception, paying particular attention to the historical and theoretical contexts within which this research was conducted. Our foci in this review fall on three principle topics: early work on the discrimination and categorization of speech sounds, more recent efforts to understand the processes and representations that subserve spoken word recognition, and research on how infants acquire the capacity to perceive their native language. Our intent is to provide the reader a sense of the progress our field has experienced over the last half century in understanding the human's extraordinary capacity for the perception of spoken language. PMID- 11881916 TI - Band-pass specific contributions of multiple generators to the auditory 40-Hz steady state potentials. AB - OBJECTIVE: The purpose of this study was to separate the composite components of the auditory 40 Hz steady-state potentials (40 Hz SSP), by differentially augmenting them with filtering at different low passes, and to compare them with their counterparts in the transient-evoked auditory middle-latency evoked potentials (AMEP). METHODS: Transient-evoked AMEP to 3.3/sec clicks and 40 Hz SSP to 40/sec clicks were recorded from 18 subjects using three orthogonally positioned electrode pairs. Each type of potentials was filtered with a 100 Hz and with a 50 Hz low pass. Equivalent dipoles of components were estimated using Three-channel Lissajous' Trajectories and compared between filter settings (50 and 100 Hz low pass) and between the transient-evoked and the steady-state potentials. RESULTS: With a band pass of 3 to 100 each period of the 40 Hz SSP consisted of a brain stem (V) and four cortical (P0 Na, Pa1 Pa2, and Nb) components. The lower-frequency components of the 40-Hz response corresponded in latency and equivalent dipole orientation to the later transient-evoked cortical AMEP components, whereas the higher-frequency components corresponded to the earlier, brain stem and primary cortical components of transient-evoked AMEP. Band-pass filtering at 3 to 50 Hz resulted in fewer components, as early brain stem and primary cortical components diminished. CONCLUSIONS: A band pass of 3 to 100 Hz for recording the 40 Hz SSP results in a composite waveform comprising of distinct brain stem and cortical generators with different orientations of their equivalent dipoles. The relative contributions of the multiple constituents are affected by the acquisition filter low pass: brain stem and primary cortical generators mostly contribute the high frequencies and later cortical contributions dominate the lower frequencies. PMID- 11881917 TI - What drives mechanical amplification in the mammalian cochlea? AB - The recent report by Peter Dallos and colleagues of the gene and protein responsible for outer hair cell somatic motility (Zheng, Shen, He, Long, Madison, & Dallos, 2000), and the work of James Hudspeth and colleagues demonstrating that vestibular stereocilia are capable of providing power that may boost the vibration of structures within the inner ear (Martin & Hudspeth, 1999), presents the tantalizing possibility that we may not be far away from answering the question what drives mechanical amplification in the mammalian cochlea? This article reviews the evidence for and against each of somatic motility as the motor, and a motor in the hair cell bundle, producing cochlear mechanical amplification. We consider three models based on somatic motility as the motor and two based on a motor in the hair cell bundle. Available evidence supports a hair cell bundle motor in nonmammals but the upper frequency limit of mammalian hearing in general exceeds that of nonmammals, in many cases by an order of magnitude or more. Only time will tell whether an evolutionary dichotomy exists (Manley, Kirk, Koppl, & Yates, 2001). PMID- 11881919 TI - The effect of birth weight and feeding of supplemental milk replacer to piglets during lactation on preweaning and postweaning growth performance and carcass characteristics. AB - The effects of piglet birth weight and liquid milk replacer supplementation of piglets during lactation on growth performance to slaughter weight was evaluated in a study carried out with 32 sows (PIC C-22) and their piglets (n = 384; progeny of PIC Line 337 sires). A randomized block design with a 2 x 2 factorial arrangement of treatments was used. Treatments were birth weight (Heavy vs Light) and liquid milk replacer (Supplemented vs Unsupplemented). The study was divided into two periods. At the start of period 1 (birth to weaning), pigs were assigned to either Heavy or Light (1.8 [SD = 0.09] vs 1.3 kg [SD = 0.07] BW, respectively, P < 0.001) litters of 12 pigs and half of the litters were given ad libitum access to supplemental milk replacer from d 3 of lactation to weaning (21 +/- 0.2 d). In period 2 (weaning to 110 kg BW), a total of 308 pigs were randomly selected from within previous treatment and sex subclasses and placed in pens of four pigs. Pigs were given ad libitum access to diets that met or exceeded nutrient requirements. Pigs in heavy litters were heavier at weaning (6.6 vs 5.7 kg BW; SE = 0.14; P < 0.001) and tended to have more pigs weaned (11.4 vs 10.9 pigs/litter; SE = 0.21; P = 0.10). After weaning, pigs in the Heavy litter had greater ADG (851 vs 796 g; SE = 6.7; P < 0.001) and ADFI (1,866 vs 1,783 g; SE = 17.6; P < 0.001), similar gain:feed (0.46 vs 0.45; SE = 0.003; P > 0.05), and required seven fewer days (P < 0.001) to reach slaughter weight compared to pigs in the Light treatment. Feeding supplemental milk replacer during lactation produced heavier pigs at weaning (6.6 vs 5.7 kg BW; SE = 0.14; P < 0.001) and tended to increase the number of pigs weaned (11.4 vs 10.9 pigs/litter; SE = 0.21; P = 0.10) but had no effect (P > 0.05) on growth performance from weaning to slaughter. However, pigs fed milk replacer required three fewer days (P < 0.01) to reach 110 kg BW. Sow feed intake and BW loss during lactation were not affected (P > 0.05) by either birth weight or milk replacer treatment. In conclusion, birth weight has a substantially greater impact on pig growth performance after weaning than increasing nutrient intake during lactation. PMID- 11881918 TI - Use of vocalic information in the identification of /s/ and /sh/ by children with cochlear implants. AB - OBJECTIVE: When a syllable such as "sea" or "she" is spoken, listeners with normal hearing extract evidence of the fricative consonant from both the fricative noise and the following vocalic segment. If the fricative noise is made ambiguous, listeners may still perceive "s" or "sh" categorically, depending on information in the vocalic segment. Do children whose auditory experience comes from electrical stimulation also display this effect, in which a subsequent segment of speech disambiguates an earlier segment? DESIGN: Unambiguous vowels were appended to ambiguous fricative noises to form tokens of the words "she," "sea," "shoe," and "Sue." A four-choice identification test was undertaken by children with normal hearing (N = 29), prelingually deaf children with the Nucleus Spectra-22 implant system using the SPEAK coding strategy (N = 13), postlingually deafened adults with the same implant system (N = 26), and adults with normal hearing (N = 10). The last group undertook the test before and after the stimuli were processed to simulate the transformations introduced by the SPEAK coding strategy. RESULTS: All four groups made use of vocalic information. Simulated processing reduced the use made by normal-hearing adults. Implanted subjects made less use than the other groups, with no significant difference between implanted children and implanted adults. The highest levels of use by implanted subjects were within one standard deviation of the mean level displayed when normal-hearing adults listened to processed stimuli. Analyses showed that the SPEAK strategy distorted formant contours in the vocalic segments of the stimuli in ways that are compatible with the errors of identification made by implanted subjects. CONCLUSIONS: Some children with implants can extract information from a following vowel to disambiguate a preceding fricative noise. The upper limit on this ability may be set by distortions introduced by the implant processor, rather than by the auditory experience of the child. PMID- 11881920 TI - Effect of wean-to-finish management on pig performance. AB - In each of three trials, 240 crossbred barrows weaned at 17 d of age (5.1 kg BW) were assigned to one of three experimental treatments based on light and heavy weight outcome groups. Experimental treatments were 1) wean-to-finish at 0.69 m2/pig and 15 pigs/pen; 2) wean-to-finish double-stocked at 0.35 m2/pig, 30 pigs per pen for 8 wk and then randomly split into two pens (either stayed in same pen or moved to new pen) for growth to slaughter at 0.69 m2/pig; and 3) nursery facility for 8 wk at 0.35 m2/pig and 15 pigs/pen followed by move to the same grow-finish facility housing wean-to-finish and double-stocked pigs and maintaining pen integrity. Beginning at 38 kg BW, diets were supplemented with either bacitracin methylenedisalicylate at 33 mg/kg to slaughter or tylosin at 44 mg/kg to 59 kg BW and 22 mg/kg thereafter. There were no trial x treatment interactions, even though there was considerable variation in health status among trials. At the end of the 56-d nursery period, wean-to-finish pigs weighed more than nursery (28.7 vs 27.7 kg; P = 0.071) and double-stocked pigs (28.7 vs 26.9 kg; P = 0.002), due to greater ADG (wean-to-finish vs nursery; P = 0.062; wean-to finish vs double-stocked; P = 0.002) and greater ADFI (wean-to-finish vs nursery; P = 0.024; wean-to-finish vs double-stocked, P = 0.002). There was no effect of treatments (P > 0.1) on ADG, feed conversion, carcass lean percentage, or lean gain during the growing-finishing period. There was also no effect of treatment (P > 0.1) on ADG or ADFI from weaning to slaughter. There was no difference (P > 0.1) between bacitracin methylenedisalicylate and tylosin for ADG, feed conversion, carcass lean percentage, or daily lean gain. These data suggest that housing 5-kg weaned pigs in fully slatted growing-finishing facilities from weaning to slaughter was not detrimental to overall performance. In this experiment, dietary additions of bacitracin methylenedisalicylate or tylosin from 38 kg BW to slaughter weight resulted in similar growth performance. PMID- 11881921 TI - Genetic antagonism between body weight and milk production in beef cattle. AB - Korean cattle have an unusually short suckling period (4 mo) due to poor milking ability, and this is a hindrance to growth of calves. Therefore, Korean cattle breeders have shown interest in genetic improvement of milking ability. In this study, body weight (birth weight, weaning weight, and yearling weight) and five daily milk yields by period in Korean cattle (Hanwoo) were analyzed using a two trait sire and maternal grandsire mixed model. The milk yields used were actually measured at sequential intervals from 1 to 4 mo after calving. Posterior means of the parameters were estimated using Gibbs sampling. Heritability estimates (0.25 to 0.26) for daily milk yield at weaning were larger than those with other periods. Genetic impact on daily milk yield, especially at weaning, was emphasized in order to lengthen the suckling period of Korean cattle. Genetic correlation estimates between BW and daily milk yield were all negative (-0.08 to -0.16 for birth weight, -0.04 to -0.21 for weaning weight, and -0.12 to -0.19 for yearling weight), whereas environmental correlation estimates were all positive (0.20 to 0.39 for birth weight, 0.34 to 0.51 for weaning weight, and 0.30 to 0.45 for yearling weight). The negative estimates of genetic correlation between weight and milk yield implied genetic antagonism between direct and maternal effects for weaning weight of beef cattle. PMID- 11881922 TI - Investigation of genotype x country interactions for growth traits in beef cattle. AB - The importance of genotype x country interactions for weaning and birth weight and postweaning gain between Argentina (AR), Canada (CA), Uruguay (UY), and the United States (US) for populations of Hereford cattle was investigated. Three sample data sets of computationally manageable sizes were formed for each trait and pairwise combination of countries to investigate possible interactions. Parameters were estimated for each sample data set via an accelerated EM-REML algorithm and multiple-trait animal models that considered either weaning or birth weight as a different trait in each country. Direct and maternal (in parentheses) weaning weight genetic correlation estimates for AR-CA, AR-UY, AR US, CA-UY, CA-US, and UY-US were 0.82 (0.80), 0.81 (0.72), 0.81 (0.79), 0.83 (0.78), 0.85 (0.82), and 0.86 (0.81), respectively. Direct and maternal (in parentheses) birth weight genetic correlation estimates were 0.92 (0.62), 0.97, (0.85), and 0.99 (0.97) for AR-CA, AR-US, and CA-US, respectively. Birth weight was not analyzed for UY due to small amounts of data. Postweaning gain in CA and US was 160-d gain, and in AR and UY 345-d gain was used. Across-country direct genetic correlations for postweaning gain were estimated for each pairwise country data set using a model that considered weaning weight as the same trait across each country, whereas postweaning gain was treated as a different trait in each country. Direct genetic correlation estimates for postweaning gain for AR CA, AR-UY, AR-US, CA-UY, CA-US, and US-UY were 0.64, 0.80, 0.51, 0.84, 0.92, and 0.83, respectively. The overall results indicate that weaning and birth weights of Hereford calves can be analyzed as the same trait in all countries with a common set of heritabilities and genetic correlations, after adjustment for heterogenous phenotypic variances across countries. Postweaning gain in CA and US can be considered as the same trait and analyzed using a single set of parameters. Postweaning gain in AR and UY should be considered as a separate trait from postweaning gain in CA and US, and postweaning gain in AR and UY can be considered as the same trait and analyzed using a common heritability, after adjustment for phenotypic variance differences between the two countries. PMID- 11881923 TI - The effects of split marketing on the physiology, behavior, and performance of finishing swine. AB - One hundred twenty 8-wk-old barrows (20.3 +/- 2.0 kg BW) were used to examine the effect of split marketing on selected behavioral, physiological and performance parameters. Pigs were assigned by weight in a randomized complete block design to one of three treatments: SM (split-marketed), six pigs/pen (1.83 m2/pig); C (control), six pigs/pen (1.83 m2/pig); or MC (modified control), three pigs/pen (3.66 m2/pig). The heaviest half of SM animals were removed 1 wk prior to marketing penmates. Control and MC animals remained in their respective groups until marketing. Animals were videotaped during the first 72 h of the study (INITIAL), 72 h prior to (PRE), and following the removal (POST) of pigs in the SM treatment to quantify maintenance behaviors and to identify socially dominant, intermediate, and submissive pigs. A blood sample was collected from each animal upon completion of INITIAL, PRE, and POST time periods to determine neutrophil:lymphocyte ratio and plasma haptoglobin, cortisol, and corticosteroid binding globulin (CBG) levels. Animals were weighed and feed disappearance was calculated biweekly. Tenth-rib backfat and area of the longissimus muscle at marketing were ultrasonically evaluated on all animals. Regardless of treatment, animals were more (P < 0.01) active (eating, standing/walking, fighting) at INITIAL than at PRE or POST times. Frequency and duration of fights per pen were less (P < 0.01) in MC than in C or SM pigs for all periods observed. Neutrophil:lymphocyte ratio, plasma haptoglobin, and CBG levels were greater (P < 0.01) during the INITIAL period than during the PRE or POST periods but did not differ between treatments. No treatment or time differences were detected in plasma cortisol levels. The MC pigs exhibited greater (P < 0.01) ADFI with poorer feed efficiency compared to C or SM pigs up to split marketing. During the POST period, both MC and SM pigs had greater (P < 0.01) ADFI with poorer (P < 0.01) feed efficiency than C pigs. The ADG was not different among animals as a result of treatment. There were no treatment differences for any of the carcass measurements. Significant differences in performance between the treatment groups could not be attributed to any physiological or behavioral measures reported here. PMID- 11881924 TI - Offstream water and trace mineral salt as management strategies for improved cattle distribution. AB - The objective of this study was to test the combined effect of offstream water and trace mineral salt on cattle distribution in a riparian meadow and its adjacent uplands. From July 15 to August 26, 1996 and 1997, three treatments were each randomly assigned to one pasture in each of three blocks. Sixty cow/calf pairs were then randomly allotted to the grazed pastures. The treatments included 1) stream access and access to offstream water and trace-mineral salt (off stream), 2) stream access and no access to offstream water or trace-mineral salt (no-offstream), and 3) ungrazed control. The response of cattle was measured through visual observations of cattle distribution, grazing activity and travel distance, cow/calf performance, and fecal deposit distribution. Distribution patterns of the cattle, measured as the distance of cattle from the stream, was characterized by a time of day x treatment x time in grazing period x year interaction (P < 0.05). No-offstream cattle began the day further from the stream than offstream cattle but consistently moved closer to the stream after the morning grazing period (0600 to 0900). Differences in distribution patterns between the two treatments were more pronounced early in the grazing period than late in the grazing period. Grazing activity, fecal deposit distribution, and travel distance of cattle were not affected by the presence of offstream water and trace-mineral salt. Cows and calves with offstream water and trace-mineral salt gained 11.5 kg and 0.14 kg/d more, respectively, than no-offstream cows and calves averaged across years (P < 0.05). Overall, cattle distribution patterns and cow/calf performance were influenced by the presence of offstream water and trace-mineral salt. Changes in distribution were most pronounced early in the grazing season. PMID- 11881925 TI - The provision of solid feeds to veal calves: I. Growth performance, forestomach development, and carcass and meat quality. AB - Growth performance, forestomach development, and carcass and meat quality of veal calves fed a milk replacer diet (Control) were compared to those obtained from calves fed the same liquid diet plus 250 g x calf(-1) x d(-1) of dried beet pulp or wheat straw. Three groups of 46 Polish Friesian calves, balanced according to initial BW, were assigned to the three dietary treatments in a fattening trial, which lasted 160 d. The provision of either solid feed did not affect the milk replacer intake. However, calves' ADG was increased (P < 0.01) only by feeding the beet pulp diet. The administration of both solid feeds improved calves' health status; calves fed solid feeds required fewer iron treatments for low hemoglobin and needed less medical treatments for respiratory or gastrointestinal diseases. In comparison to the Control calves, the provision of wheat straw and beet pulp increased iron intake throughout the fattening period by 41 and 130%, respectively. However, only calves fed beet pulp showed higher levels of hemoglobin and plasma iron concentrations (P < 0.05), whereas the same blood parameters were similar between Control calves and those fed wheat straw. At slaughter, both solid feeds led to empty forestomach weights heavier than those of Controls without reducing dressing percentage. The reticulorumen was heaviest in calves fed beet pulp, whereas wheat straw promoted omasal development. The administration of beet pulp resulted in a better carcass conformation than did the Control diet or wheat straw, but it had a detrimental effect on carcass color, which was graded as the darkest (P < 0.001). Consistent with this result, meat color of calves fed beet pulp was darker than that of Control calves and those fed wheat straw, because of the higher hematin concentration measured at the muscle level. No differences in carcass and meat color were observed between Control calves and calves fed wheat straw. The administration of solid feeds for welfare purposes does not always prevent the production of veal meat fulfilling the color standards required by the market. There is not a straight-forward relationship between a solid feed's iron content and the "redness" of veal meat, which should be related to the capability of the calves to use the iron provided by the roughage. PMID- 11881926 TI - The provision of solid feeds to veal calves: II. Behavior, physiology, and abomasal damage. AB - The aim of this study was to evaluate the effects of the addition of two roughage sources (wheat straw and beet pulp) to the milk replacer diet of veal calves, in order to reduce stress and improve animal welfare. We allocated 138 Polish Friesian male calves to three different feeding plans: a milk replacer diet (Control), 250 g/d of wheat straw in addition to the milk replacer, or 250 g/d of dried beet pulp in addition to the milk replacer. Within each feeding treatment, 16 calves were individually housed and 30 were kept in group pens (five calves/pen). Several behavioral, physiological, and health welfare indicators were monitored throughout the fattening period, which lasted for 160 d. Abnormal oral behavior around the meals was higher in Control calves (P < 0.01), while its lowest level was observed in straw-fed calves. At the beginning of the trial, chewing was higher in calves receiving solid feeds (P < 0.001), but the difference from the Control gradually decreased and disappeared at wk 13 for calves fed beet pulp and at wk 17 for those fed wheat straw. At the end of the fattening period, no differences among treatments were found in the frequency of chewing. Regardless of the diet, self-grooming decreased with age and no relationship was observed between this behavior and the presence of rumen hairballs. Cross-sucking was performed with low frequencies (from 4.70% at wk 2 to 1.05% at wk 23 around the meals, and even lower far from the meals) and was not affected by the provision of roughage. The time in contact with the bucket during the whole day was higher in Controls, whereas calves fed wheat straw maintained a lower level of this activity until the end of the trial (P < 0.01). The calves fed wheat straw spent more time in contact with the feed trough (P < 0.001) than those fed beet pulp and Control calves. No differences were found in cortisol curves due to the feeding treatment. In calves fed beet pulp, most hematological measures statistically differed from the other treatments, possibly in response to the higher iron intake and(or) to the higher hemoconcentration, probably due to the administration of beet pulp as dried feed. The incidence of abomasal ulcers and erosions was increased by the provision of the solid feeds, particularly by a structured fiber source such as straw. A roughage source able to satisfy calves' behavioral needs and to improve digestive processes without damaging the digestive apparatus still has to be identified. PMID- 11881927 TI - Degree of competition at feeding differentially affects behavior and performance of group-housed growing-finishing pigs of different relative weights. AB - The effect of competition at feeding on specific categories of growing-finishing pigs was studied in order to provide an explanation for the increased variation in performance within pens previously seen in highly competitive feeding environments. Individual feed intake and feeding behavior, performance, and health were compared for relatively small, medium, and large pigs in pens of 16. Pens contained either one or two dry feeders during a period of ad libitum eating followed by a period of restricted feeding. Computerized feeders that registered the time and amount of feed consumed by each individual were used. Seven replicate groups on each of the two treatments (a total of 224 pigs) were studied. In the pens with only one feeder, the small pigs ate less (P < 0.05) and tended to have a lower daily weight gain (P < 0.10) than the small pigs in pens with two feeders. However, they had a better feed conversion efficiency (P < 0.05). The small pigs also had a different eating pattern. A considerably larger part of their daily feed intake was consumed during the nighttime hours (P < 0.001) when only one feeder was available rather than two feeders. The large pigs ate more during the restricted period (P < 0.01) and had a lower carcass meat percentage (P < 0.05) in the pens with one feeder. Some differences in feeding behavior between the two levels of competition were seen for all categories of pigs. No difference in health was observed between pigs in the one- and two feeder pens for any size category of pigs. In conclusion, the inability of the small pigs to get access to feed in combination with overeating by the largest individuals caused the variation in performance seen within pens with a high level of competition at feeding (one feeder for 16 pigs). From a welfare point of view, feeding systems causing a high level of competition may be detrimental when considering all individual pigs in pens, even when it is possible to achieve acceptable production results on average. PMID- 11881928 TI - Daily treatment of growing foals with equine somatotropin: pathologic and endocrinologic assessments at necropsy and residual effects in live animals. AB - This experiment assessed the effects of 12 mo of daily treatment of young horses with recombinant equine somatotropin (eST) on 1) carcass and internal organ traits at necropsy and 2) residual effects in live horses for 60 d after cessation of treatment. Seven horses received eST daily at 20 microg/kg BW; seven others received vehicle (controls). Four horses from each group were killed at the end of treatment. There were few effects of eST treatment on hematologic assessments or histopathologic evaluations of internal organs. Treatment with eST increased the weights of the right adrenal gland (P = 0.090), left (P = 0.085) and right (P = 0.013) kidneys, liver (P = 0.012), tended to inrease the weights of pancreas (P = 0.082), spleen (P = 0.008), and heart (P = 0.102), and decreased (P = 0.032) somatotropin (ST) content in the adenohypophysis. Loin-eye area at the 10th rib was also greater (P = 0.01) in eST-treated horses than in controls. There was no difference (P > 0.15) between groups in left adrenal, brain, parathyroid glands, or thyroid gland weights or in 10th-rib fat thickness. In the remaining two control and three eST-treated horses (one control horse died), plasma IGF-I concentrations were higher (P = 0.001) in treated animals through d 6 after cessation of treatment and then dropped precipitously. Insulin concentrations in treated animals tended to be elevated (P = 0.08) only on d 0. There was a treatment x day interaction (P = 0.04) for plasma urea nitrogen levels, which increased in treated horses. A decrease (P < 0.05) in BW in the treated animals was observed by 21 d after treatment. There was no difference (P > 0.15) in insulin or glucose response to glucose tolerance tests given on d 0 through 60 after cessation of treatment. Overall ST response to secretagogue was reduced (P < 0.05) in eST-treated horses compared with controls. In summary, long term treatment of growing horses with eST decreased endogenous ST response to secretagogue and increased plasma IGF-I concentrations and many internal organ weights but had little effect on hematologic or histopathologic characteristics at necropsy. The effects on IGF-I concentrations were lost within 6 d, and BW in treated horses decreased within 3 wk after cessation of treatment. PMID- 11881929 TI - Beef customer satisfaction: factors affecting consumer evaluations of clod steaks. AB - An in-home beef study evaluated consumer ratings of clod steaks (n = 1,264) as influenced by USDA quality grade (Top Choice, Low Choice, High Select, and Low Select), city (Chicago and Philadelphia), consumer segment (Beef Loyals, who are heavy consumers of beef; Budget Rotators, who are cost-driven and split meat consumption between beef and chicken; and Variety Rotators, who have higher incomes and education and split their meat consumption among beef, poultry, and other foods), degree of doneness, and cooking method. Consumers evaluated each steak for Overall Like, Tenderness, Juiciness, Flavor Like, and Flavor Amount using 10-point scales. Grilling was the predominant cooking method used, and steaks were cooked to medium-well and greater degrees of doneness. Interactions existed involving the consumer-controlled factors of degree of doneness and(or) cooking method for all consumer-evaluated traits for the clod steak (P < 0.05). USDA grade did not affect any consumer evaluation traits or Warner-Bratzler shear force values (P > 0.05). One significant main effect, segment (P = 0.006), and one significant interaction, cooking method x city (P = 0.0407), existed for Overall Like ratings. Consumers in the Beef Loyals segment rated clod steaks higher in Overall Like than the other segments. Consumers in Chicago tended to give more uniform Overall Like ratings to clod steaks cooked by various methods; however, consumers in Philadelphia gave among the highest ratings to clod steaks that were fried and among the lowest to those that were grilled. Additionally, although clod steaks that were fried were given generally high ratings by consumers in Philadelphia, consumers in Chicago rated clod steaks cooked in this manner significantly lower than those in Philadelphia. Conversely, consumers in Chicago rated clod steaks that were grilled significantly higher than consumers in Philadelphia. Correlation and stepwise regression analyses indicated that Flavor Like was driving customer satisfaction of the clod steak. Flavor Like was the sensory trait most highly correlated to Overall Like, followed by Tenderness, Flavor Amount, and Juiciness. Flavor Like was the first variable to enter into the stepwise regression equation for predicting Overall Like, followed by Tenderness and Flavor Amount. For the clod steak, it is likely that preparation techniques that improve flavor without reducing tenderness positively affect customer satisfaction. PMID- 11881930 TI - Effect of dietary betaine on nutrient utilization and partitioning in the young growing feed-restricted pig. AB - The purpose of this study was to examine the effects of dietary betaine over a range of concentrations (between 0 and 0.5%) on growth and body composition in young feed-restricted pigs. Betaine is associated with decreased lipid deposition and altered protein utilization in finishing pigs, and it has been suggested that the positive effects of betaine on growth and carcass composition may be greater in energy-restricted pigs. Thirty-two barrows (36 kg, n = 8 pigs per group) were restrictively fed one of four corn-soybean meal-skim milk based diets (18.6% crude protein, 3.23 Mcal ME/kg) and supplemented with 0, 0.125, 0.25, or 0.5% betaine. Feed allotment was adjusted weekly according to BW, such that average feed intake was approximately 1.7 kg for all groups. At 64 kg, pigs were slaughtered and visceral tissue was removed and weighed. Carcasses were chilled for 24 h to obtain carcass measurements. Subsequently, one-half of each carcass and whole visceral tissue were ground for chemical analysis. Linear regression analysis indicated that, as betaine content of the diet was elevated from 0 to 0.5%, carcass fat concentration (P = 0.06), P3 fat depth (P = 0.14) and viscera weight (P = 0.129) were decreased, whereas total carcass protein (P = 0.124), protein deposition rate (P = 0.98), and lean gain efficiency (P = 0.115) were increased. The greatest differences over control pigs were observed in pigs consuming 0.5% betaine, where carcass fat concentration and P3 fat depth were decreased by 10 and 26%, respectively. Other fat depth measurements were not different (P > 0.15) from those of control pigs. In addition, pigs consuming the highest betaine level had a 19% increase in the carcass protein:fat ratio, 23% higher carcass protein deposition rate, and a 24% increase in lean gain efficiency compared with controls. Dietary betaine had no effects (P > 0.15) on growth performance, visceral tissue chemical composition, carcass fat deposition rate, visceral fat and protein deposition rates, or serum urea and ammonia concentrations. These data suggest that betaine alters nutrient partitioning such that carcass protein deposition is enhanced at the expense of carcass fat and in part, visceral tissue. PMID- 11881931 TI - Arginine catabolism in lactating porcine mammary tissue. AB - In vivo studies have shown that the uptake of plasma arginine by the lactating porcine mammary gland greatly exceeds the output of arginine in milk, but little is known about the metabolic fate of arginine in this organ. The objective of this study was to quantify arginine catabolism via arginase and nitric oxide synthase pathways in the mammary tissue of sows on d 28 of lactation. Mammary tissue slices (approximately 60 mg) were incubated at 37 degrees C for 1 h in 2 mL of Krebs bicarbonate buffer containing 0.5 or 2 mM L-[U-14C]arginine, and arginine metabolites were measured using HPLC and radiochemical techniques. Rates of arginine utilization were similar to rates of urea production. Proline, ornithine, urea, glutamate, glutamine, CO2 and polyamines (putrescine + spermidine + spermine) were formed from arginine, accounting for 46, 31, 17, 2.3, 1.5, 0.22, and 0.30%, respectively, of the metabolized arginine carbons. Relatively small amounts of arginine were utilized for nitric oxide and citrulline synthesis, with citrulline accounting for 2% of the metabolized arginine carbons. Production of all arginine metabolites increased with increasing extracellular arginine concentrations from 0.5 to 2 mM, indicating a high capacity for arginine degradation. Consistent with the metabolic findings, the activities of arginases, ornithine aminotransferase, and pyrroline-5 carboxylate reductase were high, whereas those of pyrroline-5-carboxylate dehydrogenase, ornithine decarboxylase, and nitric oxide synthases were relatively low, and there was no proline oxidase, ornithine carbamoyltransferase or pyrroline-5-carboxylase synthase activity in the mammary tissue. Our results demonstrate for the first time that proline, ornithine, and urea were the major products of arginine catabolism via the arginase pathway in lactating porcine mammary tissue and provide a biochemical basis to explain a relative enrichment of proline but a relative deficiency of arginine in sow's milk. PMID- 11881932 TI - Impact of betaine on pig finishing performance and carcass composition. AB - Two experiments were conducted to evaluate the effect of betaine supplementation of finishing diets on growth performance and carcass characteristics of swine. Experiment 1 included 288 pigs in a 2 x 2 x 3 factorial arrangement of treatments consisting of barrows and gilts of two genetic populations fed diets with 1.25 g/kg supplemental betaine from either 83 or 104 kg to 116 kg and control pigs fed betaine-devoid diets. Pigs were housed three pigs per pen with eight replicate pens per treatment. Diets were corn-soybean meal-based with 300 ppm added choline. Genetic populations differed (P < 0.05) in fat depth (2.24 vs 2.93 cm) and longissimus muscle depth (53.8 vs 49.1 mm) at 116 kg. Betaine reduced feed intake (P < 0.05); however, real-time ultrasound measurements were not affected. In Exp. 2, 400 pigs were used in a 2 x 2 x 2 factorial arrangement of treatments to evaluate the effect of sex (barrow or gilts), betaine (0 or 1 g/kg of diet), and crude protein (CP) (0.70% lysine = 12.7% CP or 0.85% lysine = 15.0% CP) when fed from 60 to 110 kg live weight. Pigs had been assigned to either a high- or low-protein feeding regimen at an average initial weight of 11.3 kg and were maintained on their respective protein levels throughout the experiment. For a 56 d period from 61.7 kg to 113.6 kg, pigs were fed diets with 300 ppm added choline. Within each protein level, pigs were randomly assigned to diets containing 0 or 1 g/kg betaine. Pigs were group-housed (four to five pigs per pen). Pig weight and feed intake were recorded every 28 d. Real-time ultrasound measurements were recorded initially and at d 28 on 64 pigs, and on all pigs prior to slaughter. Growth rate was fastest and feed intake greatest for barrows (P < 0.05) and for pigs receiving 12.7% crude protein. A crude protein x betaine interaction (P < 0.05) was observed from d 28 to 56 with pigs fed the 15% CP diet growing fastest when supplemented with 1 g/kg betaine, and pigs receiving the 12.7% CP diet growing fastest when the diets contained 0 g/kg betaine. Gilts more efficiently (P < 0.05) converted feed into body weight gain, as did pigs receiving the 12.7% CP diet (P < 0.05). Longissimus muscle area and fat measurements were unaffected by betaine or dietary protein on d 28. However, by d 56 betaine reduced average fat depth in barrows (P < 0.05; 3.21 vs 3.40 cm), but not in gilts. Betaine may be more effective at altering body composition in barrows than in gilts. PMID- 11881933 TI - Effect of spray-dried plasma and lipopolysaccharide exposure on weaned pigs: I. Effects on the immune axis of weaned pigs. AB - A study was conducted with 20 weaned barrows (14 d, 4.98 +/- .21 kg) to determine the effect of spray-dried plasma (SDP) on the pig's immune response to a lipopolysaccharide (LPS) challenge. After weaning, pigs were fed a diet containing 0 or 7% SDP for 7 d. On d 6 postweaning, all pigs were fitted with a jugular catheter. On d 7 postweaning, the pigs were given an i.p. injection of either saline or LPS (150 microg/kg BW) followed by a 3-h blood collection every 15 min. Following blood collection, all pigs were killed and tissue was collected for mRNA analysis. Additionally, the small intestine was collected for measurement of villus height, crypt depth, and villus height:crypt depth ratio (VCR) at three sites (25, 50, and 75% of the total length). Feeding SDP resulted in reduced (P < 0.05) mRNA expression of tumor necrosis factor-alpha (TNF-alpha) and interleukin-1beta (IL-1beta) mRNA in the adrenal gland, spleen, hypothalamus, pituitary gland, and liver. Additionally, expression of IL-6 mRNA was reduced (P < 0.05) in the spleen and pituitary gland for pigs fed SDP. For pigs fed the diet with SDP, LPS administration did not affect (P > 0.10) cytokine mRNA expression, whereas LPS reduced expression of TNF-alpha mRNA in the spleen and IL-1beta mRNA in the adrenal gland, spleen, and thymus for pigs fed the diet without SDP. For pigs fed the diet with SDP, LPS caused serum TNF-alpha to increase 150-fold compared to a 60-fold increase for pigs fed the diet without SDP. Similarly, interferon-gamma (IFN-gamma) increased 110-fold for pigs fed the diet with SDP compared to a 16-fold increase for pigs fed the diet without SDP. For pigs fed the diet with SDP, LPS caused major villus atrophy, whereas for pigs fed the diet without SDP, LPS had no effect on intestinal morphology. These results demonstrate that the basal activation of the immune system appears to be less for pigs fed the diet with SDP compared to pigs fed the diet without SDP after weaning. Additionally, for pigs fed the diet with SDP, there appeared to be an overresponse of the immune system following LPS administration, which resulted in major damage to the mucosa of the gastrointestinal tract. PMID- 11881934 TI - Effect of spray-dried plasma and lipopolysaccharide exposure on weaned pigs: II. Effects on the hypothalamic-pituitary-adrenal axis of weaned pigs. AB - A study was conducted with 20 weaned barrows (14 d, 4.98 +/- 0.21 kg) to determine the effect of feeding spray-dried plasma (SDP) after weaning on the pig's stress response to a lipopolysaccharide (LPS) challenge. After weaning, pigs were fed a diet containing 0 or 7% SDP for 7 d. On d 6 after weaning, all pigs were nonsurgically fitted with a jugular catheter. On d 7 after weaning, the pigs were given i.p. injections of either saline or LPS (150 microg/kg BW) followed by serial blood collection every 15 min for a 3-h period. Following the 3-h blood collection, all pigs were killed and tissue was collected for mRNA analysis. Pig weight on d 7 after weaning was not affected by dietary treatment (P > 0.21). Pigs fed the diet with SDP had lower (P < 0.05) levels of hypothalamic corticotropin-releasing hormone (CRH) mRNA, pituitary gland CRH receptor mRNA, and adrenal gland adrenocorticotropin-releasing hormone (ACTH) receptor mRNA. Dietary treatment did not affect pituitary gland proopiomelanocortin (POMC) mRNA. No effect of LPS treatment was observed in any of the mRNA levels examined. For both serum ACTH and cortisol, there was a significant diet x LPS treatment interaction (P < 0.01) such that both the ACTH and cortisol responses to the LPS challenge were greater in the pigs fed the diet with SDP than in the pigs fed the diet without SDP. For pigs given the saline injection, diet did not affect basal serum cortisol concentration; however, basal serum ACTH concentration was lower in those pigs fed the diet with SDP (P < 0.0001). A diet x LPS treatment interaction (P < 0.024) was observed for adrenal gland mRNA expression for steroidogenic acute regulatory (StAR) protein such that the LPS-induced increase in StAR mRNA was greater in the pigs fed SDP than in pigs fed the diet without SDP. These results demonstrate that pigs fed a diet with SDP have an increased activation of the pituitary-adrenal axis following an LPS challenge compared to pigs fed a diet without SDP. PMID- 11881935 TI - Effect of inoculants on whole-crop barley silage fermentation and dry matter disappearance in situ. AB - Whole-crop barley harvested at the mid-dough stage was ensiled in 3-L laboratory silos either directly (at 30.7% DM) or after wilting (37.8% DM), and with or without application of Inoculant A, B, or D. Each inoculant contained multiple strains of Lactobacillus plantarum and Enterococcus faecium. Two silos per treatment were opened on d 1, 3, 7, 15, and 47 for silage analysis. Wilted silages had higher (P < 0.05) pH than unwilted silages, and Inoculant B (unwilted crop) and Inoculants A, B, and D (wilted crop) decreased (P < 0.05) silage pH compared to the controls. Reducing sugars concentration was 36% lower (P < 0.05), on average, in the wilted than in the unwilted silages. Silages prepared with Inoculants B or D had lower (P < 0.05) concentrations of reducing sugars than the control. Wilting of the barley crop further increased the difference between inoculated and control silages (wilting x inoculation P < 0.05). Neither wilting nor silage inoculants affected concentrations of nonprotein N, ammonia N, or free amino acid N in silage. Wilting did not affect the concentration of lactic acid bacteria (LAB) in the silages. Across DM levels, inoculant-treated silages had larger (P < 0.01) populations of LAB than did the uninoculated controls (7.1 x 10(9) vs 2.3 x 10(9) cfu/g silage DM). Wilting resulted in lesser (P < 0.05) silage lactic acid concentration than the directly ensiled crop. At both DM levels, lactic acid concentration was higher (P < 0.001) in inoculated silages than in controls. The in situ soluble and potentially degradable fractions of silage DM were not affected by wilting or inoculant treatment. The rate of degradation of the potentially degradable silage DM was 35% lower (P < 0.05) in wilted than in unwilted silage. As a result, the calculated effective degradability of silage DM was lower (P < 0.001) in wilted than in unwilted silage. Inoculants did not influence the rate of degradation or effective degradability of silage DM in the rumen. Whole-crop barley ensiled at approximately 30% DM (without wilting) contained higher concentrations of soluble sugars and lactic acid and had higher ruminal degradability of DM than wilted silage (38% DM). Although inoculants did not improve DM degradability of barley silage, lower terminal pH and increased concentrations of lactic acid may improve aerobic stability upon feed-out. PMID- 11881936 TI - In vitro gas production measurements to evaluate interactions between untreated and chemically treated rice straws, grass hay, and mulberry leaves. AB - In vitro gas production was measured to investigate associative effects of untreated and chemically treated rice straw (RS) and of RS and grass hay or mulberry leaves (ML). The RS was treated with NaOH, urea, or (NH4)HCO3. Cumulative gas production was recorded at 2, 4, 6, 8, 12, 16, 24, 36, 48, 60, 72, and 96 h of incubation, and the Gompertz function was used to describe the kinetics of gas production. Treatment with NaOH, urea, and NH4HCO3 increased (P < 0.05) gas production at 48 h by 55, 52, and 37% and the maximum rate of gas production of RS from 0.64 to 1.51, 1.27, and 1.13 mL/h, respectively. The inclusion of treated straws, hay, and ML in a mixture with RS at increasing proportions (25, 50, 75, 100%) elevated cumulative gas production and its rate. Maximum gas production was not different between the RS and its mixtures with the treated straws, but inclusion of hay into RS or NH4HCO3-treated RS increased the maximum gas production. Associative effects were defined as the difference between the observed gas production for the mixtures and the sum of the individual component feeds. The NaOH-treated RS, included at higher proportions, had positive associative effects at all times of incubation < 96 h; effects were negative or absent at 96 h. No effects were observed with mixtures of RS with urea-treated RS. The NH4HCO3-treated RS mixture tended to have negative associative effects at all three levels. Positive associative effects (P < 0.05) were observed for both RS and NH4HCO3-treated RS at almost all inclusion levels of hay or ML after 12 to 96 h of incubation. The response was more pronounced with ML than with hay. Associative effects generally declined with duration of incubation. We conclude that positive associative effects on in vitro gas production occurred more consistently when RS was incubated in mixtures with hay or ML than when incubated in mixtures with chemically treated RS. PMID- 11881937 TI - Net absorption and utilization of nitrogenous compounds across ruminal, intestinal, and hepatic tissues of growing beef steers fed dry-rolled or steam flaked sorghum grain. AB - Our objectives were to determine effects of grain processing on splanchnic (gut tissues and liver) N metabolism and whole-body N balance by growing steers and to ascertain the relative contributions of ruminal and intestinal tissues to net absorption and utilization of N-containing nutrients. Seven beef steers (348 kg initial BW), surgically implanted with appropriate catheters, were fed diets containing 77% steam-flaked (SF) or dry-rolled (DR) sorghum grain. Blood flows and net output or uptake of ammonia N, urea N, and alpha-amino N (estimate of amino acids) were measured across portal-drained viscera (PDV or gut tissues) and intestinal, ruminal, hepatic, and splanchnic tissues (PDV + hepatic). The experimental design was a crossover between DR and SF diets, with six samplings of blood at 2-h intervals on 2 d for each steer. Nitrogen intake (139 +/- 3 g/d), output in urine (43 +/- 2 g/d), and retention (40 +/- 3 g/d) were similar for both processing treatments. When steers were fed SF sorghum compared to DR sorghum, N retention as a percentage of N intake was numerically greater (P < 0.12), output of fecal N was numerically lower (P < 0.13), and urinary urea N was lower (P < 0.04). For SF vs DR, net uptake of alpha-amino N by liver was higher (P < 0.04; 20 vs 9 g/d) and was numerically lower (P < 0.16) for ruminal tissues (15 vs 33 g/d). Feeding steers SF compared to DR tended to increase net transfer (cycling) of blood urea N to PDV (57 vs 41 g/d; P < 0.07), increased cycling to intestinal tissues (15 vs 6 g/d; P < 0.05), and numerically increased transfer to ruminal tissues (42 vs 32 g/d; P < 0.12) but did not alter other net output or uptake of N across splanchnic tissues. Total urea N transfer (blood + saliva) was similar for both treatments. Net uptake of alpha-amino N by ruminal tissues was about 30% of the net amount of alpha-amino N absorbed across the intestinal tissues. In summary, most of the blood urea N cycled from the liver to gut tissues was transferred to ruminal tissues for potential microbial protein synthesis, and the net ruminal utilization of alpha-amino N was about 30% of that absorbed from intestinal tissues. Feeding growing steers SF compared to DR sorghum diets numerically increased whole-body N retention (percentage of N intake) by about 15% and tended to increase transfer of blood urea N to the gut by about 40%, which could increase the supply of high-quality microbial protein for absorption. PMID- 11881938 TI - The biohydrogenation of linoleamide in vitro and its effects on linoleic acid concentration in duodenal contents of sheep. AB - Previous studies showed that oleamide was protected from ruminal biohydrogenation and increased 18:1(n-9) concentration in milk when fed to lactating dairy cows. To appraise whether this protection extended to linoleamide, a rumen in vitro experiment was conducted to determine biohydrogenation of linoleamide followed by two sheep experiments to evaluate whether linoleamide could increase 18:2 (n-6) concentration in duodenal contents. Treatments for the in vitro and sheep studies consisted of three diets containing no added lipid (control), linoleic acid, or linoleamide. Lipids were added at 10% (DM basis) of the in vitro substrate (ground grass hay). The three substrates were incubated with mixed ruminal microbes in triplicate, and 5 mL of culture contents was taken at 0, 24, and 48 h for analysis of 18:2 (n-6) concentration by gas chromatography. The concentrations of 18:2 (n-6) (corrected for 18:2 (n-6) in the control cultures) at 0, 24, and 48 h were 2.51, 0.38, and 0.11 mg/5 mL for the linoleic acid cultures compared to 2.10, 1.35, and 1.08 mg/5 mL for the linoleamide cultures. Compared to linoleic acid, the cultures containing linoleamide had higher 18:1 (n 9) and lower concentrations of biohydrogenation products including trans-18:1 and 18:0. Three sheep with duodenal cannulas were fed the three diets in two separate 3 x 3 Latin squares each with 2-wk periods. The two squares only differed in the amount of added lipid (1.5 vs 5% of the ration DM). When the lipids were added at 1.5% of the ration DM, they had little effect on duodenal 18:2 (n-6) concentration (2.8, 3.6, and 4.3 mg/g DM for the control, linoleic acid, and linoleamide treatments, respectively). At 5% of the ration DM, both lipid supplements increased duodenal 18:2 (n-6) concentration over the control diet with a greater response observed for linoleamide (2.5, 12.2, and 16.8 mg/g DM for the control, linoleic acid, and linoleamide treatments, respectively). This study demonstrates reduced biohydrogenation of linoleamide based on its ability to maintain a higher concentration of 18:2 (n-6) in ruminal cultures and in duodenal contents of sheep compared to free linoleic acid. PMID- 11881939 TI - Rapid communication: Thirty-eight polymorphic microsatellite markers for mapping in rainbow trout. PMID- 11881940 TI - Canadian nursing research in the new millennium. PMID- 11881942 TI - I'm not a "basabasa" woman: an explanatory model of HIV illness in Ghanaian women. AB - Ghana continues to experience an increase in the rate of infection with the human immunodeficiency virus (HIV), with more new infections occurring in women than in men. Prevailing views of health and illness, including indigenous knowledge and traditional beliefs, are an important component of the broad context of disease transmission. Participatory action research was used to explore the explanatory model of HIV illness of 31 seropositive Ghanaian women. Also interviewed were 5 HIV seropositive men, 2 traditional healers, 8 nurses, and 10 professionals, individually and in focus groups, to reflect on the women's comments and the themes emerging from the data. In this article, the women's beliefs about HIV illness will be discussed and their views about the etiology, pathophysiology, symptomology, course of illness, and methods of treatment for their illness will be described. Findings illustrate areas of divergence and convergence between traditional and biomedical explanations of, and treatment for, HIV illness. The necessity for health professionals, particularly nurses, to understand individual and community perceptions about HIV illness is highlighted by the study findings. PMID- 11881941 TI - God in control: women's perspectives on managing HIV infection. AB - Women with HIV face a number of challenges in living with this chronic, life threatening illness: economic, physical, social, and emotional. When discussing their illness the importance these women place on having a strong spiritual life is a consistent theme. In this study, women were asked to describe and explain what spirituality meant to them and how they used it in living with HIV. The results indicate that, in opposition to what some contemporary providers fear HIV positive women use their spiritual life to enhance the care prescribed by providers, rather than using their faith to avoid mainstream sources of care. In addition, women related the importance of spirituality in dealing with everyday life. Knowledgeable providers can incorporate discussions of spirituality in their care of women with HIV and, in the process, potentially improve the therapeutic results of their HIV-specific care. PMID- 11881943 TI - An observational study of the development of a nurse-patient relationship. AB - The purpose of this exploratory descriptive case study was to identify features of nurse-patient interactions (NPI) in the development of a nurse-patient relationship (NPR). Observations of interactions in a cancer treatment unit included 60 videotaped, sequential, naturally occurring NPIs involving one dyad over a 3-day period. A microanalysis of the interactions was conducted using qualitative ethological methods. The analysis focused on identifying important recurring behavioral clusters and characteristic patterns of behavior that comprised these clusters. Important features of NPIs in the development of an NPR were identified and described in detail. The active and complementary roles of both participants in this process and the contribution of social exchange, trust, and humor are highlighted. The findings illustrate the importance of continuity of nursing care if NPRs are to be used to their fullest extent to address complex patient care needs. PMID- 11881944 TI - Impact of comprehensive nurse-managed transitional care. AB - The purpose of this study was to test the effect of nurse-managed transitional care on the quality of care and functional ability of individuals following discharge from subacute units. Registered nurses employed on subacute units in a skilled nursing facility provided the nurse-managed transitional care. Using a quasi-experimental design, data were collected on admission to the subacute unit, at the time of discharge, 1 week following discharge, and 3 months following discharge on 242 treatment and comparison participants. The treatment group participants' overall function and quality of the care environment were significantly higher than the comparison group at 1 week and 3 months following discharge. Participants did not differ significantly on basic activities of daily living or number of readmissions. PMID- 11881945 TI - The Kim Alliance Scale: development and preliminary testing. AB - The development and preliminary testing of the Kim Alliance Scale (KAS) is described. The KAS was developed to measure the quality of the therapeutic alliance from the patient's perspective, including patient empowerment. By triangulating literature findings and qualitative data, a conceptual framework was derived with four dimensions of alliance: collaboration, communication, integration, and empowerment. Using the 48-item KAS, 68 nurses evaluated their alliance (as patients) with their own health care providers. Standard psychometric procedures were used to test for reliability and validity. The KAS was reduced to 30 items having an internal consistency reliability coefficient alpha of .94 and split-half alphas of .87 and .91. Initial support was found for convergent and divergent validity. The KAS may be a useful tool for measuring therapeutic alliance, identifying the foci for interventions to improve the alliance, and improving treatment outcomes. PMID- 11881947 TI - The research time line. PMID- 11881946 TI - Nursing communication: advocacy for the patient or physician? AB - Communication among nurses, patients, and physicians is a key component of effective health care. In addition to communication with patients, nurses directly or indirectly influence physician-patient communications. This secondary analysis examined registered nurses' interactions with a simulated patient regarding what the physician had told the patient about the reason for hospitalization. Taped interviews (N = 86) were transcribed and content analyzed to classify nurses' approaches to assessment and intervention. The second researcher coded 10% of the transcripts to ensure satisfactory interrater consistency. Major patterns of nursing communication were the following: assessing what the physician had told the patient (85%), encouraging clarification with the physician (62%), encouraging a second opinion, and defending the physician's competence (9%). Findings support literature suggesting that nurses mediate and clarify communications between the patient and the physician. Patient advocacy was also illustrated; however, a small number of nurses advocated most clearly for the physician. PMID- 11881948 TI - Family caregiving in long-term care facilities. AB - Family caregiving within institutional settings is a neglected area of research In an era of cost containment, more and more is being asked of families of institutionalized persons. This article presents the results of a study designed to investigate family caregiving following the admission of an elderly relative to a long-term care facility. Beginning with a brief summary of findings from previous research, the article reports on visiting and task performance by families within the long-term care facility. Continuing with descriptions of families' satisfaction with institutional care and their care-related learning needs, the article concludes with recommendations for policy and practice. PMID- 11881949 TI - Counseling Hypertensive patients: an observational study of 21 public health nurses. AB - This study observed the public health nurse's and the patient's activity level during blood pressure measurement and the kind of nonpharmacological treatment that was given. Using the Nurse Practitioner Rating Form, three structured observations were made of 21 public health nurses at their offices at health care centers. The nurses were randomly selected from 22 health care centers in Southern Sweden. The public health nurses used nonpharmacological treatment at 18 out of 63 visits, mainly diet and physical activity. The nonpharmacological conversation had a psychosocial aspect at 15 observations. During the visits, most of the facts and advice concerned somatic aspects of health promotion. Almost all patients were asked about their medication. At more than half of the observations, the nurses and the patients met at the same medium or high communication level. The nurses need training and information about nonpharmacological treatment to practice health promotion in hypertension care. PMID- 11881950 TI - Using simulations and standardized patients in intervention research. AB - Caregiving is both an art and a science; both knowledge and skills are necessary to improve the quality of care that professionals provide. Researchers are often limited in their assessment of skill performance due to inadequate and unrealistic measurement options. Simulation using standardized patients offers researchers an objective method to evaluate skill performance. This article describes the use of simulations by researchers in their quest to measure changes in violence prevention skills after an intervention program given to nurse assistants working in long-term care facilities. PMID- 11881951 TI - Quality of life in persons living with HIV. AB - This study investigated the effects of selected variables on quality of life (QOL) in persons living with HIV. Eighty-one HIV-positive adults completed a demographic questionnaire, the Sickness Impact Profile, the Center for Epidemiologic Studies Depression Scale, and a fatigue visual analog scale. Blood samples were collected for CD4+, CD8+, and CD16+ Lymphocyte numbers. Based on an expanded psychoneuroimmunology model, hierarchical multiple regression analysis was used to determine the extent to which four blocks of variables predicted variance in quality of life. These included sociodemographics, depression, immune status, and fatigue. Findings revealed that work status, depression, and fatigue predicted 58% of the variance in QOL in persons with HIV Findings support the need for routine assessment and management of depression and fatigue in persons living with HIV. PMID- 11881952 TI - Body piercing and tattooing perspectives. AB - This descriptive, correlational study surveyed 79 pierced and/or tattooed participants to determine reasons why people pierce and tattoo their bodies and to assess participants' knowledge of health risks involved in body alteration procedures. Participants queried represented a wide age range-between 19 and 55. Results showed that participants perceived few health risks involving piercing and tattooing and desired additional piercings and/or tattoos. Individual expression was an important body alteration motivation for both piercing and tattooing. These findings underscore the importance of health care professionals' maintaining nonjudgmental attitudes about those who alter their bodies, there by facilitating important health education concerning related health risks. Suggestions for nursing applications are discussed. PMID- 11881953 TI - Adult patients' postoperative pain descriptions and responses to the Short-Form McGill Pain Questionnaire. AB - This study examined the clinical usefulness of the Short-Form McGill Pain Questionnaire (SF-MPQ). Thirty postoperative patients were asked to describe their postoperative pain and were then administered the SF-MPQ. Eighteen (60%) used exact SF-MPQ sensory or affective words or synonyms to describe their postoperative pain during the interview. These results provide further evidence of the clinical relevance of the SF-MPQ sensory and affective scales. Pain descriptions by patients that go beyond pain intensity descriptions may communicate more precise information about the pain and lead to more effective pain interventions. Patients with difficulty describing their pain might be assisted by using the SF-MPQ. PMID- 11881954 TI - The Future of Dentistry Report aims at helping dentists and patients. PMID- 11881955 TI - Calcium crisis bodes ill for bones. PMID- 11881956 TI - Embezzlement causes more than financial damage. PMID- 11881957 TI - Why is ergonomics an issue in dentistry? PMID- 11881959 TI - Ergonomic risk factors associated with clinical dentistry. AB - Ergonomics has formed an integral aspect of dental education at the University of British Columbia since the early 1980s. However, studies continued to indicate that dentists are at risk for developing musculoskeletal problems. This provided the impetus for a study of the risk factors associated with these problems. The data analyzed from 421 survey respondents in British Columbia indicate that indeed dentists are experiencing musculoskeletal pain and discomfort. However, the data also suggest that dentists can recognize and identify their own postures, practicing positions, and the equipment usage patterns that are associated with increased risks of experiencing musculoskeletal pain and discomfort. Such recognition is the first critical step to avoiding or neutralizing ergonomic habits and work environment layouts that might otherwise unnecessarily shorten professional clinical careers. PMID- 11881958 TI - Ergonomics and the dental office: an overview and consideration of regulatory influences. AB - Nearly 2 million workers suffer from musculoskeletal disorders each year. These problems are caused by repetitive, awkward, or stressful motions. Dental health care workers are susceptible to these types of injuries. This article will discuss state and federal programs to control job-related injuries and relate the regulations to dental practice. PMID- 11881960 TI - Ergonomic benefits of surgical telescope systems: selection guidelines. AB - A longstanding myth holds that chronic neck and back discomfort or pain is a necessary evil of practicing dentistry. The use of properly selected surgical telescopes and co-axial illumination headlights has been demonstrated to prevent or in some cases eliminate chronic neck and back pain. This paper will discuss the ergonomic benefits of surgical telescopes and co-axial illumination lights, the recent advances made in surgical telescope technology, and practical guidelines for selecting telescopes. PMID- 11881961 TI - Injury prevention for the practice of dentistry. AB - There is an abundance of dental professionals with work-related pain and dysfunction. Dentistry poses a huge challenge because of the ergonomics of dental work. The biggest risk factors are the awkward prolonged seated postures with no back support and the limited range of motion and isometric muscle contraction created by working in a confined area, namely the mouth. The following manuscript offers exercises that constitute a preventive program to target the typical problem areas of the dental professional--the neck, lower back and wrist/hand. PMID- 11881962 TI - Meeting of the minds. PMID- 11881963 TI - S-enantiomer of the selective serotonin reuptake inhibitor citalopram compared with racemate citalopram (S+R). PMID- 11881964 TI - The scientific conflict. PMID- 11881965 TI - Pathology and pathophysiology of drug-induced arterial injury in laboratory animals and its implications on the evaluation of novel chemical entities for human clinical trials. AB - In toxicology studies, drug-induced arterial injury in laboratory animals continues to be a pressing issue of concern, particularly to those engaged in the discovery and development of novel therapies intended for human use. The concern is justifiably magnified because, currently, there is no unequivocal biochemical marker of arterial injury and/or toxicity in animals or man. Therefore, in laboratory animals used for toxicology studies a precise description of arterial lesions in terms of location, distribution and morphologic character is necessary so that a correlation can be drawn between structural damage and derangement of specific cardiovascular functions. The critical nature of the latter cannot be over-emphasized because this will provide a basis for understanding the mechanism of toxicity, the pathogenesis of the lesion and assessment of human risk. However, in the decision making process, utilization of pattern recognition must be supported by rigorous scientific investigations aimed at establishing a link, where possible, between the deranged cardiovascular function and alterations in cellular, biochemical and molecular events. Conceivably, engagement of the molecular pharmacology target initiates a series of interactive cascades among cellular and non-cellular arterial components that culminate in organ damage. Therefore, any investigative mechanistic studies aimed at understanding the initiation and development of arterial lesions in laboratory animals must make a conscientious attempt to identify and characterize the molecular target of toxicity. PMID- 11881966 TI - Preclinical comparison of ebastine and other second generation H1-antihistamines. AB - The present study was performed to compare the properties of ebastine--the long duration of antiallergic effect and less penetration to the CNS--with those of other H1-antihistamines. Passive cutaneous anaphylactic reaction was induced and the dye leakage from the skin measured after oral administration of the various H1-antihistamines in guinea pigs. The H1-antihistamines examined inhibited passive cutaneous anaphylactic reactions, with ED50 values of 1.55-5.77 mg/kg administered orally. Evaluation at doses close to the ED50 values determined that the rank order of the various H1-antihistamines for the duration of antiallergic effects, calculated from the AUC, was as follows: ebastine>cetirizine> or =oxatomide=loratadine=epinastine. The inhibition of [3H]-mepyramine binding to the cortical membrane was examined ex vivo after oral administration of the drugs in rats. Ketotifen as a positive control of sedative antihistamine, oxatomide, cetirizine, ebastine and epinastine dose-dependently inhibited the [3H] mepyramine binding to rat cortical membranes. However, ebastine and epinastine did not show 50% [3H]-mepyramine binding inhibition even at 100 mg/kg orally In conclusion, ebastine was shown to be a potent and long-lasting H1-antihistamine with less effect to the CNS. Consequently, in conjunction the two experimental models used in this study--passive cutaneous anaphylactic reaction in guinea pigs and ex vivo [3H]-mepyramine binding to rat cortical membrane--may be important to estimate the duration of antiallergic effects of drugs and to detect their sedative effects, which are important indicators in the development of new antiallergic drugs. PMID- 11881967 TI - Lead induces endothelium- and Ca2+-independent contraction in rat aortic rings. AB - The contractile effect of lead on rat aortic rings was examined. Lead (0.1-3.1 mM) elicited concentration-dependent but endothelium-independent contractions, which were unaffected by prazosin (1 microM). The contractile effects of lead were similar when the aortic rings were bathed either in the absence or presence of external Ca2+. Lanthanum (1 mM) but not verapamil (I pM) inhibited the lead contractions; hence non-L-calcium channels are involved in such effect. In addition, lead induced contractions on aortic rings incubated in Ca2+-free EGTA containing solution for 70 min., an experimental condition in which intracellular Ca2+-stores are depleted. Finally, the contractile effect of lead was not modified by calphostin C (an inhibitor of protein kinase C). In conclusion, the present results suggest that in rat aorta, the lead-induced contraction is independent of extra- and intracellular calcium stores. In addition, the effect of lead is independent of either catecholamines or protein kinase C. It is likely that in rat aorta, lead enters into the smooth muscle cells through non-L-calcium channels, and when acting like calcium on the contractile machinery it produces contraction. The differences observed between our results and those obtained by other authors may indicate that the mechanism of the contractile effect of lead varies among the different blood vessels. PMID- 11881968 TI - High NaCl intake decreases both flow-induced dilation and pressure-induced myogenic tone in resistance arteries from normotensive rats: involvement of cyclooxygenase-2. AB - The effect of high NaCl diet on resistance arteries is not yet fully documented. In order to assess the effect of NaCl on myogenic tone and flow-induced dilation independent of arterial blood pressure change, we used normotensive rats which did not develop hypertension upon high NaCl intake. Normotensive Wistar Kyoto Rats received a high (8%) or a normal NaCl diet (0.4%). Mesenteric resistance arteries (150 microm, internal diameter) were cannulated in an arteriograph to allow perfusion of arteries under controlled pressure and flow. Pressure-induced myogenic tone was lower in the high NaCl group than in the control group. Cyclooxygenase inhibition with indomethacin and (N-(2-cyclohexyloxy)-4-nitro phenyl)-methanesulphonamide, 1 micromol/l) NS 398 (specific cyclooxygenase-2 inhibitor) similarly decreased myogenic tone in rats fed high NaCl but had no effect in those fed a normal NaCl diet. Flow-induced dilation was decreased in the high NaCl group. Inhibition of nitric oxide synthesis with N(G)-nitro-L arginine methyl ester decreased flow-induced dilation in both groups. Indomethacin and NS 398 did not change flow-induced dilation. As shown by immunofluorescence COX-2 was present in the endothelium of arteries from rats with a high NaCl diet but not in those fed a normal NaCl diet. Thus, chronic high NaCl intake decreased both flow-induced dilation and myogenic tone in resistance arteries. The chronic high NaCl did not affect the participation of nitric oxide on flow-induced dilation, but induced the expression of cyclooxygenase-2, which participates in myogenic tone. These results suggest that high NaCl changes flow and pressure mechanosensing processes and strengthen the hypothesis that sodium ions have an important role in both pressure and flow-mechanotransduction in vascular cells. PMID- 11881969 TI - Trimeresurus flavoviridis (habu snake) venom induces human erythrocyte lysis through enzymatic lipolysis, complement activation and decreased membrane expression of CD55 and CD59. AB - Trimeresurus flavoviridis (habu snake) bites can be fatal to man because of its virulent venom, which is clinicopathologically classified as haemorrhagic, necrotic, and haemolytic toxins. Trimeresurus flavoviridis venom causes lysis of human erythrocytes in conditions where plasma is present as well as in plasma free conditions in a dose-dependent manner. The haemolytic process requires Ca2+ and Mg2+ ions in the solution. Additionally, the venom initiates activation of the human complement cascade as evidenced by C3a and C5a releases, complement consumption indicated by CH50 and formation of soluble membrane attack complex. The insertion of membrane attack complex into the erythrocyte membranes is morphologically identified by electronmicroscopy. Immunofluorescence analysis reveals that incubation of erythrocytes with the venom decreased cell-surface expression of CD55 (decay accelerating factor) and CD59 (protectin), which renders erythrocyte more vulnerable to adherent C3 and C5 convertases and to polymerization of C9 into membranes, and may enhance autologous complement mediated haemolysis triggered by the venom. Our data demonstrate that Trimeresurus flavoviridis venom induces haemolysis in the presence of plasma by three distinct mechanisms, direct lipolysis through PLA2 activity, activation of the human complement system, and cleavages of CD55 and CD59 from erythrocyte membranes. PMID- 11881970 TI - Inhibition of rat platelet 5-hydroxytryptamine uptake by chlorpyrifos and carbaryl. AB - The organophosphate insecticide chlorpyrifos and the carbamate insecticide carbaryl were investigated in adult male rats in terms of their effects on the activity of brain monoamine oxidase-A (MAO-A) activity and on the platelet uptake of 5-hydroxytryptamine (5-HT). The activities of brain acetylcholinesterase (AChE) and plasma butyrylcholinesterase (BuChE) were also determined. For each compound two different dosage regimens were employed. In the acute study, chlorpyrifos or carbaryl was administered at a single intraperitoneal dose of 100 mg/kg or 50 mg/kg, respectively In the subacute study, chlorpyrifos was injected at a daily dose of 20 mg/kg for 7 days, while carbaryl was given at a daily dose of 10 mg/kg for 14 days. Acute chlorpyrifos administration produced a 85.01% inhibition of AChE and a 43.4% inhibition of BuChE but had no effect on MAO-A activity and 5-HT uptake. In contrast, subacute chlorpyrifos exposure caused a 94.96% inhibition of AChE and a 85.8% inhibition of BuChE and, also, elicited a significant (35.02%) reduction in the platelet uptake of 5-HT. MAO-A activity was not affected. Acute carbaryl administration produced a 56.38% AChE inhibition and a 55.95% BuChE inhibition and also caused a significant (26.36%) decrease in 5-HT uptake but no change in MAO-A. Subacute carbaryl exposure failed to affect significantly any of the biochemical parameters determined. Interference with the 5-HT system by chlorpyrifos and carbaryl could contribute to the toxicity of these pesticides. PMID- 11881971 TI - Behavioural effects in rats after prenatal exposure to dearomatized white spirit. AB - The purpose of the study was to investigate the potential developmental neurotoxicity of the widely used organic solvent, white spirit. Rats (Mol:WIST) were exposed to 0 or 800 ppm dearomatized white spirit for 6 hr per day on gestation days 7-20. Developmental and neurobehavioural effects in the offspring were investigated using a test battery including assessment of physical development, reflex ontogeny, motor function, motor activity and, learning and memory. No significant effects were recorded on motor function and the activity in Open Field. In the initial learning period (age 1 month), the performance in a Morris water maze was similar in exposed and control animals. When testing for memory at the age of 2 months, the exposed male offspring used more time to locate the hidden platform. After platform relocation, impaired cognitive function was revealed in the exposed females. At the age of 5 months, learning and memory deficits were observed in exposed offspring. The differences were not related to poorer swimming capabilities, because swim speeds were similar to control values. The results show that prenatal exposure to 800 ppm white spirit caused long-lasting learning and memory deficits in rats. PMID- 11881972 TI - Effects of CCK2 receptor blockade on growth parameters in gastrointestinal tract and pancreas in rats. AB - Gastrin has a growth-promoting effect on the oxyntic mucosa of the stomach but has been claimed also to affect other parts of the gastrointestinal tract and pancreas. This report describes the effects of the cholecystokinin, (CCK2) receptor antagonists YM022 and YF476 on various growth parameters in the gastrointestinal tract and pancreas of the rat. YM022 and YF476 were given subcutaneously in doses known to produce maximum and sustained CCK2 receptor blockade. The body weight was not affected. However, the oxyntic mucosal weight, thickness and protein and DNA contents were reduced by 15-20% already within 1-2 days and by about 30% after 4-8 weeks of CCK2 receptor blockade. Hence, the response of the oxyntic mucosa to CCK2 receptor blockade was in the form of hypotrophy (reduced protein content) and hypoplasia (reduced DNA content). There were no obvious effects of CCK2 receptor blockade on the intestine or pancreas (nor on liver, kidney or thyroid). The proton pump inhibitor omeprazole was used to induce hypergastrinaemia and was given with or without YM022. Omeprazole treatment for 4 weeks increased the oxyntic mucosal weight and thickness by 15 20%. YM022 prevented these effects. We conclude that while elevated circulating gastrin levels, acting on CCK2 receptors, exert a growth-promoting effect on the oxyntic mucosa (but not elsewhere), normal serum gastrin levels exert a mucosa preserving effect. PMID- 11881973 TI - Tetrandrine prevents tissue inhibitor of metalloproteinase-1 messenger RNA expression in rat liver fibrosis. PMID- 11881974 TI - Novel aspects of dopamine oxidative metabolism (confounding outcomes take place of certainties). AB - Abstract: Understanding dopamine (DA) oxidative metabolism allows to get a deeper insight into neurologic and psychiatric disorders featured by an altered DA neurotransmission as well as developing appropriate therapeutic strategies. Oxidative DA deamination is carried out by two highly conserved isoenzymes: monoamine oxidase (MAO) A and B; these isoenzymes both metabolize DA to dihydroxyphenylacetaldehyde (DOPALD), which, in turn, is converted to dihydroxyphenylacetic acid (DOPAC). In the past twenty years most studies on MAO activity were performed using brain dialysis in freely moving rats and measuring DA and DOPAC levels after administration of specific MAO inhibitors. This led to concepts on DA metabolism grounded on a single brain area (striatum) investigated, almost exclusively, in a single animal species (rat). These experiments were based on measurement of striatal levels of DOPAC which represents the indirect product of MAO activity. At present, the specific role of each MAO isoform appears to differ significantly depending on varying experimental conditions such as measuring the direct product of DA metabolism. In particular, recent studies allowed the estimate of the first metabolite (DOPALD) formed by MAO, showing that DOPAC levels do not necessarily reflect MAO activity. Again, the relative contribution of the two MAO iso forms in sustaining DA metabolism varies considerably, depending on the animal species and the specific brain area (either striatum or substantia nigra) under investigation. In this article we will briefly review these concepts in light of new evidence derived from innovative approaches: improved in vivo analysis of direct MAO metabolic products; measurement of oxidative metabolism in different parts of the DA nigrostriatal pathway; measurement of MAO activity in various animal species including MAO knock-out mice. PMID- 11881975 TI - Autoxidation and toxicant-induced oxidation of lipid and DNA in monkey liver: reduction of molecular damage by melatonin. AB - Melatonin, the main secretory product of the pineal gland, is a free radical scavenger and antioxidant which protects against oxidative damage due to a variety of toxicants. However, there is little information regarding melatonin's antioxidative capacity in tissues of primates. In this study we examined the protective effects of melatonin in monkey liver homogenates against lipid damage that occurred as a result of autoxidation or that induced by exogenous addition of H202 and ferrous iron (Fe2+). Additionally, we tested melatonin's protective effect against oxidative damage to DNA induced by chromium(III) (CrIII) plus H202. The levels of malondialdehyde and 4-hydroxyalkenals were assayed as an index of lipid peroxidation, and the concentrations of 8-hydroxydeoxyguanosine (8 OHdG) as an endpoint of oxidative DNA damage. The increases in malondialdehyde+4 hydroxyalkenals concentrations as a consequence of autoxidation or after the addition of H202 plus Fe2+ to the homogenates were time-dependent. The accumulation of these damaged products due to either auto-oxidative processes or induced by H202 and Fe2+ were significantly reduced by melatonin in a concentration-dependent-manner. The levels of 8-OHdG were elevated in purified monkey liver DNA incubated with a combination of CrCl3 plus H2O2. This rise in oxidatively damaged DNA was prevented by 10 microM concentration of melatonin. Also, melatonin reduced the damage to DNA that was caused by auto-oxidative processes. These findings in monkey liver tissue document the ability of melatonin to protect against oxidative damage to both lipid and DNA in primate tissue, as observed previously in rodent tissue. The findings provide support for the use of melatonin as suitable agent to reduce damage inflicted by free radical species in primates. PMID- 11881976 TI - A new method to evaluate in vitro myelotoxicity of antitumour agents in the first steps of drug development. AB - Research focused on the development of new anticancer agents has been based mainly on the assessment of the antitumour activity. This yields a large number of newly developed drugs endowed with good antitumour properties, but heavy side effects on myelopoiesis. In this work, we validate a new method potentially useful to assess myelotoxic effect of newly developed agents. The proposed technique uses peripheral blood CD34+ cells as source of haematopoietic progenitors. These cells are grown in liquid culture in the presence of cytokines able to induce differentiation versus the three main lineages. Doxorubicin, carboplatin and topotecan served as reference drugs to investigate the accuracy of the technique. The three drugs mimick the effects reported in vivo. Doxorubicin and carboplatin produce a specific effect toward erythropoietic and thrombopoietic lineages, respectively, and topotecan a three-lineage toxicity. An advantage of the technique is the possibility to further investigate myelotoxicity. Here, we assessed differentiation markers in CD34+ cells to evaluate if the three drug treatments can affect the process of differentiation. Data show that the drug treatments were unable to modulate the expression of the selected differentiation markers in the surviving population. We propose this method as an innovative tool to score the myelotoxic effect of compounds in the first steps of drug development to further develop those compounds with the best ratio between activity and myelotoxic effects. Moreover, the fact that the method is performed in liquid phase allows its optimisation in a conventional "high throughput system". PMID- 11881977 TI - Role of monoamine oxidase inhibition and monoamine depletion in fenfluramine induced neurotoxicity and serotonin release. AB - The role of both monoamine synthesis and monoamine oxidase inhibition in mediating the fenfluramine-induced damage to serotonin neurones was examined; as pretreatment agents, both alpha-methyl-para-tyrosine (AMPT) and parachlorophenylalanine (PCPA) were used to deplete dopamine and serotonin, respectively, while clorgyline and deprenyl were used to inhibit monoamine oxidase types A and B. While both AMPT and deprenyl did not alter fenfluramine induced serotonin or 5-hydroxyindoleacetic acid (5-HIAA) depletion in any area, PCPA did partially reduce the serotonin depletion in the hippocampus and hypothalamus. Although pretreatment with clorgyline did not significantly alter fenfluramine-induced serotonin depletion, it did produce a 65% mortality rate in animals treated with both drugs. Both PCPA and clorgyline significantly increased the depletion of striatal 5-HIAA concentration consequent to fenfluramine; however, these drugs also produced a long-term depletion of striatal 5-HIAA when administered alone, therefore, the changes seen after the coadministration with fenfluramine may be viewed as additive. Finally, acute PCPA pretreatment attenuated the rapid rise in 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (homovanillic acid) induced by fenfluramine, and acute clorgyline reversed the drop in serotonin and rise in 5-HIAA induced by fenfluramine. These results indicate that the rapid increase in dopamine activity induced by fenfluramine is partially dependent on serotonin concentration and release and that the mechanism of fenfluramine-induced toxicity is unlike that of the other substituted amphetamines. PMID- 11881978 TI - Neurochemical and behavioural effects of chronic aldicarb administration in rats. AB - Changes in neurochemical parameters, behavioural and memory performances due to aldicarb (a carbamate pesticide) treatment was investigated after chronic oral administration in rats. Rats received water, or aldicarb 1 ppb (= 0.12 g/kg/day), aldicarb 10 ppb (= 1.2 g/kg/day) and aldicarb 100 ppb (= 12 g/kg/day) for 4 months. The locomotor and explorative activities were reduced, and aldicarb significantly decreased brain AChE activity while an increase was found in the passive avoidance and water-maze performance. In the striatum, aldicarb 10 ppb and 100 ppb, significantly reduced DOPA but not DOPAC concentrations, while the 10 ppb and 100 ppb doses significant increased DA levels. Aldicarb did not affect DA, DOPA and DOPAC levels in the accumbens. The neuropharmacologic effects of chronic dosing with aldicarb, AChE inhibition and dopaminergic modulation could be useful for treatment of memory deficits related to CNS disorders. PMID- 11881979 TI - Cyanoside chloride and chromocarbe diethylamine are more effective than vitamin C against exercise-induced oxidative stress. AB - Exercise generates free radicals only when it is exhaustive. Free radicals are involved in tissue damage caused by exercise. Antioxidant vitamins (vitamin C and E) and other antioxidants such as coenzyme Q, and N-acetyl cysteine prevent muscle damage and decrease muscle fatigue. The main aim of this paper was to test the possible protective effect of two new antioxidants, cyanoside chloride and chromocarbe diethylamine, on the oxidative stress generated by exhaustive exercise. The antioxidants were given to rats daily (50 mg/kg) in drinking water for 30 days. Blood oxidized glutathione/ reduced glutathione ratio, and plasma malondialdehyde levels were determined as indexes of oxidative stress. Plasma creatine kinase, alanine-aminotransferase and lactate dehydrogenase activities were used as markers of muscle damage. Both cyanoside chloride and chromocarbe diethylamine were more effective than vitamin C in the prevention of glutathione oxidation in blood. Furthermore, cyanoside chloride and chromocarbe diethylamine partially prevented muscle damage. Chromocarbe diethylamine was the most effective compound in the prevention of exercise-induced lipid peroxidation (malondialdehyde) in plasma. PMID- 11881980 TI - Spasmolytic effects of three harmala alkaloids on guinea-pig isolated trachea. AB - The present study examined and compared the spasmolytic effects of 3 harmala alkaloids, harmine, harman, and harmaline, on carbachol-, histamine-, and KCl induced contractions of guinea-pig isolated tracheal preparations. All 3 compounds relaxed the tracheal preparations contracted by these spasmogens with similar or different EC50 values, harmine being the most potent one. The cumulative concentration-response curves of all 3 compounds for carbachol-induced contraction were shifted to the right by propranolol (1 microM) pretreatment, indicating the involvement of the activation on the beta-adrenoceptors. All 3 compounds shifted the concentration-response curves of carbachol to the right in a parallel manner with the pA2 values comparable with their relaxation EC50 values, indicating a competitive antagonism at the muscarinic receptors. Receptor binding assays indicated that all 3 compounds interacted with lung muscarinic receptors (Ki = 11-13 microM), histamine H1 receptors (Ki = 27-107 microM), and beta2-adrenoceptors (Ki = 20-51 microM). Therefore, in addition to their actions on receptor-linked and voltage-dependent Ca2+ channels as reported in other types of smooth muscle, the present study suggests that the actions on muscarinic receptors, histamine H1 receptors, and beta2-adrenoceptors are also involved in their spasmolytic effects on airway smooth muscles. PMID- 11881981 TI - Single- and multiple-dose pharmacokinetics, kidney tolerability and plasma protein binding of tenoxicam in renally impaired patients and healthy volunteers. AB - The 20 mg single-dose and 12 days repeated-dose pharmacokinetics of tenoxicam and the 5-OH-tenoxicam metabolite have been evaluated in healthy volunteers and two groups of patients with different degree of renal impairment, in total 20 persons. Concomitantly, the plasma protein binding of tenoxicam and the effects of treatment on renal function were evaluated. No differences were found between the investigated groups in the pharmacokinetics of total tenoxicam and the 5-OH metabolite did not interfere either with the pharmacokinetics or with the plasma protein binding of tenoxicam. A positive correlation was found between an increase in the free fraction (% F) of tenoxicam in plasma and a decrease in the plasma elimination half-life in the low creatinine clearance group (40-20 ml/min.) both after the single-dose and at steady-state. At steady-state, a non linear correlation was demonstrated between a decrease in the urinary excretion of the 5-OH metabolite and a decrease in creatinine clearance from 130 to 20 ml/min. An increase in the plasma level of the 5-OH metabolite by three times was found in the low creatinine clearance group as compared to healthy subjects. 14C Impurities of tenoxicam, as low as 1.2%, were shown to greatly influence the determination of the plasma protein binding (equilibrium dialysis) of the highly protein-bound tenoxicam due to a non-binding ability of the impurities to plasma proteins. No significant changes in renal parameters were found during the study. It can be concluded that the pharmacokinetics and plasma protein binding of tenoxicam and the pharmacokinetics of the 5-OH-tenoxicam metabolite are increasingly changed in subjects with a creatinine clearance below 40 ml/min. A decreased binding of tenoxicam to plasma proteins in low clearance patients is probably the reason for a faster elimination of tenoxicam in this group rather than a higher intrinsic hepatic metabolic activity. This study conducted in a low number of patients did not bring forward any new data indicating any adverse effects of tenoxicam on renal function. PMID- 11881982 TI - Haem metabolic aspects on nitric oxide formation in endotoxaemia. PMID- 11881983 TI - Effects of cyclic peptide (Pro-Leu-Gly)2 on calcium signaling in isolated myometrial cells from pregnant rat. PMID- 11881984 TI - Serine-threonine protein phosphatase inhibitors: development of potential therapeutic strategies. PMID- 11881985 TI - 6,7-Dihydro-2-benzothiophen-4(5H)-ones: a novel class of GABA-A alpha5 receptor inverse agonists. AB - Nonselective inverse agonists at the benzodiazepine binding site on the GABA-A chloride ion channel enhance cognitive performance in animals but cannot be used in the treatment of cognitive disorders because of anxiogenic and convulsant side effects. We have identified a novel series of GABA-A alpha5 receptor ligands during our search for alpha5 receptor inverse agonists as potential cognition enhancers. In particular, 6,6-dimethyl-3-(2-hydroxyethyl)thio-1-(thiazol-2-yl) 6,7-dihydro-2-benzothiophen-4(5H)-one (26) has been identified as a functionally selective GABA-A alpha5 inverse agonist. PMID- 11881986 TI - 3-(1H-Pyrrol-1-yl)-2-oxazolidinones as reversible, highly potent, and selective inhibitors of monoamine oxidase type A. AB - 3-(1H-Pyrrol-1-yl)-2-oxazolidinones 1aminus signi have been synthesized as pyrrole analogues of toloxatone (Humoryl), an antidepressant agent belonging to the 3-phenyl-2-oxazolidinone class, and their monoamine oxidase (MAO) type A and B inhibitory activities have been evaluated. The majority of 1aminus signi showed inhibitory activity against the A isoform of the enzyme higher than that exerted against the MAO-B, the sole exception being the (S)-5-aminomethylderivative 1d. (R)-5-Methoxymethyl-3-(1H-pyrrol-1-yl)-2-oxazolidinone 1b, the most potent among test derivatives, was 78-fold more potent than toloxatone. PMID- 11881987 TI - Synthesis and X-ray studies of chiral allosteric modifiers of hemoglobin. AB - This study was designed to investigate the effect of chirality on the allosteric activity of a series of Hb allosteric modifiers. The chiral analogues were based on the lead compound (4), JP7, [1-[4-(((3,5 dimethylanilino)carbonyl)methyl)phenoxy]cyclopentanecarboxylic acid] with different D- and L-amino acids conjugated to the JP7 acid moiety. The D-isomers were the most potent in vitro effectors in Hb solutions as well as with whole blood. In general, this study demonstrated that the chirality of extended amino acid side chains in JP7 conjugates plays an important role in observed degree of allosteric activity. The binding site interactions for four analogues were determined by single crystallographic diffraction studies. Conclusions show that the chiral configuration of some of the D-isomers enable the effectors to bind with a greater number of interactions with the protein residues. D- and L-isomers with equivalent or near equivalent allosteric activity did not show any significant differences or interactions between their amino acid side chains and the protein. The most potent effectors, in vitro, were compounds 15 and 19, D isomers of leucine and phenylalanine, respectively. Compounds 21, 22, 30, and32 were more potent in vitro in Hb solutions than JP7. PMID- 11881988 TI - Ribose-modified nucleosides as ligands for adenosine receptors: synthesis, conformational analysis, and biological evaluation of 1'-C-methyl adenosine analogues. AB - 1'-C-Methyl analogues of adenosine and selective adenosine A(1) receptor agonists, such as N-[(1R)-1-methyl-2-phenylethyl]adenosine ((R)-PIA) and N(6) cyclopentyladenosine, were synthesized to further investigate the subdomain that binds the ribose moiety. Binding affinities of these new compounds at A(1) and A(2A) receptors in rat brain membranes and at A(3) in rat testis membranes were determined and compared. It was found that the 1'-C-methyl modification in adenosine resulted in a decrease of affinity, particularly at A(1) and A(2A) receptors. When this modification was combined with N(6) substitutions with groups that induce high potency and selectivity at A(1) receptors, the high affinity was in part restored and the selectivity was increased. The most potent compound proved to be the 1'-C-methyl analogue of (R)-PIA with a K(i) of 23 nM for the displacement of [(3)H]CHA binding from rat brain A(1) receptors and a > 435-fold selectivity over A(2A) receptors. In functional assays, these compounds inhibited forskolin-stimulated adenylate cyclase with IC(50) values ranging from 0.065 to 3.4 microM, acting as full agonists. Conformational analysis based on vicinal protonminus signproton J-coupling constants and molecular mechanics calculations using the MM2 force field proved that the methyl group on C1' in adenosine has a pronounced impact on the furanose conformation by driving its conformational equilibrium toward the north, gamma+, syn form. PMID- 11881990 TI - A comprehensive study of the active site residues of DT-diaphorase: rational design of benzimidazolediones as DT-diaphorase substrates. AB - A series of quinone substrates were modeled into the active site of human DT diaphorase and minimized. Correlation of these models with the substrate specificity k(cat)/K(m) provided insights into the structural requirements of quinone substrates. The W105, F106, and H194 residues can influence the position of the quinone substrate in the active site resulting in formation of one of the two possible Michael anions resulting from hydride transfer from FADH(2). Electron withdrawing groups on the substrate can stabilize these anions resulting in excellent substrate specificity. Inspection of models indicated that the W-105 and F-106 residues form parallel walls that will accommodate large polycyclic substrates. Thus excellent polycyclic substrates of DT-diaphorase were designed. However, the placement of tetrahedral centers on these polycyclic substrates interfered with the W-105 and the F-106 residues resulting in their exclusion from the active site. The histidine (H194) residue permits recognition of substrate enantiomers as a result of hydrogen bonding interactions. As a result of this study, it will be possible to design poor to excellent substrates of DT diaphorase and take advantage of varying levels of this enzyme in histologically different cancers. PMID- 11881989 TI - Synthesis and biological evaluation of 2-substituted 3beta-tolyltropane derivatives at dopamine, serotonin, and norepinephrine transporters. AB - A series of eight 2-substituted 3-tolyltropane derivatives were synthesized, and the in vitro and in vivo biological activities as dopamine uptake inhibitors were determined. From the in vitro structure-activity data, it is apparent that a tolyl group in the 2-position, independent of the stereochemical attachment to the tropane ring system, provided compounds (9-12, 14) that exhibit high-affinity binding at the dopamine transporter (DAT). Although a slight stereochemical preference in binding affinity at the DAT was observed for the 2beta-(R)-alcohol 10 over the 2beta-(S)-isomer 11, no significant differences in behavioral effects were observed. Furthermore, despite a relatively low potency of 10 for the inhibition of dopamine uptake compared to its affinity for the DAT, its behavioral profile did not vary significantly from cocaine. These data indicate that a behavioral characterization of compounds is a critical feature of efforts to discover pharmacological treatments for cocaine abuse. PMID- 11881992 TI - Designing small, nonsugar activators of antithrombin using hydropathic interaction analyses. AB - Conformational activation of antithrombin is a critical mechanism for the inhibition of factor Xa, a proteinase of the blood coagulation cascade, and is typically achieved with heparin, a polyanionic polysaccharide clinically used for anticoagulation. Although numerous efforts have been directed toward the design of better activators, a fundamental tenet of these studies has been the assumed requirement of an oligo- or a polysaccharide backbone. We demonstrate here a concept that small nonsaccharidic nonpolymeric molecules may be rationally designed to interact with and activate antithrombin for enhanced inhibition of factor Xa. The rational design strategy is based on a study of complexes of natural and mutant antithrombins with heparin-based oligosaccharides using hydropathic interaction (HINT) technique, a quantitative computerized tool for analysis of molecular interactions. A linear correlation was observed between the free energy of binding for antithrombinminus signoligosaccharide complexes and the HINT score over a wide range of approximately 13 kcal/mol, indicating strong predictive capability of the HINT technique. Using this approach, a small, nonsugar, aromatic molecule, (minus sign)-epicatechin sulfate (ECS), was designed to mimic the nonreducing end trisaccharide unit DEF of the sequence specific heparin pentasaccharide DEFGH. HINT suggested a comparable antithrombin-binding geometry and interaction profile for ECS and trisaccharide DEF. Biochemical studies indicated that ECS binds antithrombin with equilibrium dissociation constants of 10.5 and 66 microM at pH 6.0, I 0.025, and pH 7.4, I 0.035, respectively, that compare favorably with 2 and 80 microM observed for the natural activator DEF. ECS accelerates the antithrombin inhibition of factor Xa nearly 8-fold demonstrating for the first time that conformational activation of antithrombin is feasible with appropriately designed small nonsugar organic molecules. The results present unique opportunities for de novo activator design based on this first-generation lead. PMID- 11881991 TI - PRO_SELECT: combining structure-based drug design and array-based chemistry for rapid lead discovery. 2. The development of a series of highly potent and selective factor Xa inhibitors. AB - In silico screening of combinatorial libraries prior to synthesis promises to be a valuable aid to lead discovery. PRO_SELECT, a tool for the virtual screening of libraries for fit to a protein active site, has been used to find novel leads against the serine protease factor Xa. A small seed template was built upon using three iterations of library design, virtual screening, synthesis, and biological testing. Highly potent molecules with selectivity for factor Xa over other serine proteases were rapidly obtained. PMID- 11881993 TI - Development of 2,4-diaminopyrimidines as antimalarials based on inhibition of the S108N and C59R+S108N mutants of dihydrofolate reductase from pyrimethamine resistant Plasmodium falciparum. AB - The reduced binding of pyrimethamine to Ser108Asn (S108N) mutants of parasite dihydrofolate reductase (DHFR), which forms the basis of resistance of Plasmodium falciparum to pyrimethamine, is largely due to steric constraint imposed by the bulky side chain of N108 on Cl of the 5-p-Cl-phenyl group. This and other S108 mutants with bulky side chains all showed reduced binding to pyrimethamine and cycloguanil. Less effect on binding to some bulky mutants was observed for trimethoprim, with greater flexibility for the 5-substituent. S108N DHFR also binds poorly with other pyrimethamine derivatives with bulky groups in place of the p-Cl, and the binding was generally progressively poorer for the double (C59R+S108N) mutant. Removal of the p-Cl or replacement with m-Cl led to better binding with the mutant DHFRs. Pyrimethamine analogues with unbranched hydrophobic 6-substituents showed generally good binding with the mutant DHFRs. A number of compounds were identified with high affinities for both wild-type and mutant DHFRs, with very low to no affinity to human DHFR. Some of these compounds show good antimalarial activities against pyrimethamine-resistant P. falciparum containing the mutant DHFRs with low cytotoxicity to three mammalian cell lines. PMID- 11881995 TI - Substituted pentacyclic carbazolones as novel muscarinic allosteric agents: synthesis and structure-affinity and cooperativity relationships. AB - Two series of pentacyclic carbazolones, 22 and 23, have been synthesized utilizing a facile intramolecular Dielsminus signAlder reaction and are allosteric modulators at muscarinic acetylcholine receptors. Their affinities and cooperativities with acetylcholine and the antagonist N-methylscopolamine (NMS) at M(1)minus signM(4) receptors have been analyzed and compared. All of the synthesized compounds are negatively cooperative with acetylcholine. In contrast, the majority of the compounds exhibit positive cooperativity with NMS, particularly at M(2) and M(4) receptors. The subtype selectivity, in terms of affinity, was in general M(2) > M(1) > M(4) > M(3). The largest increases in affinity produced by a single substitution of the core structure were given by the 1-OMe (22b) and 1-Cl (22d) derivatives. The position of the N in the ring did not appear to be important for binding affinity or cooperativity. Two compounds 22y and 23i, both trisubstituted analogues, were the most potent compounds synthesized, with dissociation constants of 30minus sign100 nM for the M(2) NMS liganded and unliganded receptor, respectively. The results indicate that the allosteric site, like the primary binding site, is capable of high-affinity interactions with molecules of relatively low molecular weight. PMID- 11881994 TI - Substituted diphenyl sulfides as selective serotonin transporter ligands: synthesis and in vitro evaluation. AB - A series of diphenyl sulfide derivatives substituted at the 1-, 2'-, and 4' positions has been synthesized and evaluated for their in vitro affinities at the dopamine, serotonin (SERT), and norepinephrine transporters. The examination of K(i) values revealed that most of these derivatives have high affinity and selectivity for the SERT. Moreover, substitutions at these positions differently influence the SERT binding: (i) The nature of the substituent linked at the 1 position critically influences the SERT affinity. (ii) Functions containing heteroatom at the 2'-position afford compounds with high SERT affinity. (iii) The nature of the substituent at the 4'-position slightly influences the SERT affinity whereas steric effect markedly decreases the SERT affinity. From this series, the most SERT selective derivatives (such as 8b, 8c, and 8g) are now evaluated for their potential as positron emission tomography imaging agents when labeled with carbon-11. PMID- 11881996 TI - Synthesis of new covalently bound kappa-carrageenan-AZT conjugates with improved anti-HIV activities. AB - This paper describes the first covalent synthesis of kappa-carrageenan-3'-azido 3'-deoxythymidine (AZT) conjugates. A succinate diester spacer was used to covalently couple AZT onto kappa-carrageenan, resulting in a tripartite prodrug. Two methods (UV and radioactive counting) are described and validated to determine the AZT loading onto the kappa-carrageenan carrier. This polymeric carrier, through its own intrinsic anti-HIV activity, is expected to act not only as a drug delivery agent but also as an anti-HIV agent. Synergism between the two drugs (kappa-carrageenan and AZT) was demonstrated when MT-4 cells were preincubated with the kappa-carrageenan-AZT conjugate prior to HIV-1-infection. A threshold of AZT loaded onto the kappa-carrageenan was required to achieve this synergistic effect. Such kappa-carrageenan-AZT conjugates could be of great therapeutic interest because these conjugates, which contain a low AZT concentration, present improved anti-HIV activities relative to free AZT. Moreover, kappa-carrageenan is a well-tolerated biopolymer, already used in the food industry. PMID- 11881997 TI - Carbocyclic dinucleoside polyphosphonates: interaction with HIV reverse transcriptase and antiviral activity. AB - Carbocyclic alpha, gamma-bis(nucleoside)-5,5'-triphosphonates and alpha, delta bis(nucleoside)-5,5'-tetraphosphonates (Ap4A and Gp4G) analogues were shown to be a new type of terminating substrate of HIV reverse transcriptase. They effectively inhibited the DNA synthesis catalyzed by this enzyme in model cell free systems, but their antiviral activity both in Rat1 fibroblast cell culture bearing MLV reverse transcriptase and in HIV-infected MT-4 cells was low. When a liposome delivery system was used, the antiviral efficacy of the compounds under study was increased. PMID- 11881998 TI - First non-ATP competitive glycogen synthase kinase 3 beta (GSK-3beta) inhibitors: thiadiazolidinones (TDZD) as potential drugs for the treatment of Alzheimer's disease. AB - Glycogen synthase kinase 3 beta (GSK-3beta) has a central role in Alzheimer's disease (AD). Selective inhibitors which avoid tau hyperphosphorylation may represent an effective therapeutical approach to the AD pharmacotherapy and other neurodegenerative disorders. Here, we describe the synthesis, biological evaluation, and SAR of the small heterocyclic thiadiazolidinones (TDZD) as the first non-ATP competitive inhibitor of GSK-3beta. Their synthesis is based on the reactivity of sulfenyl chlorides. In GSK-3beta assays, TDZD derivatives showed IC(50) values in the micromolar range, whereas in other protein kinases assays they were devoid of any inhibitory activity. SAR studies allowed the identification of the key structural features. Finally, a possible enzymatic binding mode is proposed. PMID- 11881999 TI - Novel 4-anilinoquinazolines with C-7 basic side chains: design and structure activity relationship of a series of potent, orally active, VEGF receptor tyrosine kinase inhibitors. AB - We have previously shown that 4-anilinoquinazolines can be potent inhibitors of vascular endothelial growth factor (VEGF) receptor (Flt-1 and KDR) tyrosine kinase activity. A novel subseries of 4-anilinoquinazolines that possess basic side chains at the C-7 position of the quinazoline nucleus have been synthesized. This subseries contains potent, nanomolar inhibitors of KDR (median IC(50) 0.02 microM, range 0.001-0.04 microM), which are comparatively less potent vs Flt-1 tyrosine kinase (median IC(50) 0.55 microM, range 0.02-1.6 microM). The compounds also retain some inhibitory activity against the tyrosine kinase associated to the endothelial growth factor receptor (EGFR) (median IC(50) 0.2 microM, range 0.075-0.8 microM) but demonstrate selectivity vs that associated to the FGF receptor 1 (median IC(50) 2.5 microM, range 0.9-19 microM). This selectivity profile is also evident in a growth factor-stimulated human endothelial cell (HUVEC) proliferation assay (i.e., inhibition of VEGF > EGF > FGF), with inhibition of VEGF-induced proliferation being achieved at nanomolar concentrations (median IC(50) 0.06 microM). Further examination of compound 2 (ZD6474) in recombinant enzyme assays revealed excellent selectivity for the inhibition of KDR tyrosine kinase (IC(50) 0.04 microM) vs the kinase activity of erbB2, MEK, CDK-2, Tie-2, IGFR-1R, PDK, PDGFRbeta, and AKT (IC(50) range: 1.1 to >100 microM). Anilinoquinazolines possessing basic C-7 side chains exhibited markedly improved aqueous solubility over previously described anilinoquinazolines possessing neutral C-7 side chains (up to 500-fold improvement at pH 7.4). In addition, aqueous solubility of the neutral fraction present at pH 7.4 of the basic subseries of anilinoquinazoline proved to be higher than that of the neutral analogue 1 (ZD4190). Oral administration of representative compounds to mice (50 mg/kg) produced plasma levels between 0.2 and 3 microM at 24 h after dosing. Our development candidate 2 demonstrated a very attractive in vitro profile combined with excellent solubility (330 microM at pH 7.4) and good oral bioavailability in rat and dog (> 80 and > 50%, respectively). This compound demonstrated highly significant, dose-dependent, antitumor activity in athymic mice. Once daily oral administration of 100 mg/kg of compound 2 for 21 days inhibited the growth of established Calu-6 lung carcinoma xenografts by 79% (P < 0.001, Mann Whitney rank sum test), and substantial inhibition (36%, P < 0.02) was evident with 12.5 mg/kg/day. PMID- 11882000 TI - Structure-activity relationships of 2'-fluoro-2',3'-unsaturated D-nucleosides as anti-HIV-1 agents. AB - We studied the structure-activity relationships of a series of 2'-fluoro-2',3' unsaturated D-nucleosides against HIV-1 in human peripheral blood mononuclear (PBM) cells. The target compounds 10-21 and 28-33 were prepared by N glycosylation of the acetate 4, which was readily prepared from 2,3-O isopropylidene-D-glyceraldehyde in five steps. Among the newly synthesized nucleosides, 2-amino-6-chloropurine (11), adenine (14), inosine (16), guanine (18), 2,6-diaminopurine (20), and 5-fluorocytosine (30) derivatives were found to exhibit interesting anti-HIV activities with EC(50) values of 4.3, 0.44, 1.0, 2.6, 3.0, and 0.82 microM, respectively. The implications for drug resistance of the titled nucleosides with respect to lamivudine-resistant variants (M184V) were also examined, and no significant cross-resistance with the variants was observed with the D-series. PMID- 11882001 TI - Development of long-acting dopamine transporter ligands as potential cocaine abuse therapeutic agents: chiral hydroxyl-containing derivatives of 1-[2-[bis(4 fluorophenyl)methoxy]ethyl]-4-(3-phenylpropyl)piperazine and 1-[2 (diphenylmethoxy)ethyl]-4-(3-phenylpropyl)piperazine. AB - In our search for long-acting agents for the treatment of cocaine abuse, a series of optically pure hydroxylated derivatives of 1-[2-[bis(4 fluorophenyl)methoxy]ethyl]-4-(3-phenylpropyl)piperazine (1) and 1-[2 (diphenylmethoxy)ethyl]-4-(3-phenylpropyl)piperazine (2) (GBR 12909 and GBR 12935, respectively) were synthesized and evaluated in vitro and in vivo. The enantiomers of the 2-hydroxylated analogues displayed substantial enantioselectivity. The S enantiomers displayed higher dopamine transporter (DAT) affinity and the R enantiomers were found to interact at the serotonin transporter (SERT) with higher affinity. The two-carbon spacer between the hydroxyl group and the piperazine ring was essential for enantioselectivity, and the length of the alkyl chain between the phenyl group and the piperazine ring influenced binding affinity and selectivity for the DAT and SERT. Phenylethyl analogues had a higher binding affinity for the SERT and a weaker affinity and selectivity for the DAT than the corresponding phenylpropyl analogues. Thus, (S) (+)-1-[4-[2-[bis(4-fluorophenyl)methoxy]ethyl]piperazinyl]-3-phenylpropan-2-ol (6) displayed the highest affinity to the DAT, and (S)-(+)-1-[4-[2 (diphenylmethoxy)ethyl]piperazinyl]-3-phenylpropan-2-ol (8) had the highest selectivity. The latter (8) is one of the most DAT selective ligands known. In accord with the in vitro data, 6 showed greater potency than 7 in elevating extracellular dopamine levels in a microdialysis assay and in inhibiting cocaine maintained responding in rhesus monkeys. PMID- 11882002 TI - Syntheses and antiproliferative activities of new rebeccamycin derivatives with the sugar unit linked to both indole nitrogens. AB - The synthesis of new rebeccamycin derivatives, in which the carbohydrate moiety is attached to both indole nitrogens, is described. The newly synthesized compounds were tested for their abilities to block the cell cycle of murine leukemia L1210 cells and their in vitro antiproliferative activities against four tumor cell lines (murine L1210 leukemia and human HT29 colon carcinoma, A549 non small-cell lung carcinoma, K-562 leukemia). Their biological activities are compared with those of the parent compound rebeccamycin. Some of the new compounds exhibit potent antiproliferative activities, either against the four cell lines or mostly the two leukemias (L1210 and K-562 cell lines). The 3,9 diformyl analogue 9 was selective toward L1210 cells, whereas the 3,9-dibromo 16 was strongly cytotoxic toward the four cell lines tested. Nonselective compound 16 and 3,9-dinitro 13, which exhibited selectivity toward leukemia tumor cell lines, were selected for in-depth evaluation, including in vivo experiments. PMID- 11882003 TI - Synthesis and mode of action of hydrophobic derivatives of the glycopeptide antibiotic eremomycin and des-(N-methyl-D-leucyl)eremomycin against glycopeptide sensitive and -resistant bacteria. AB - Des-(N-methyl-D-leucyl)eremomycin was obtained by Edman degradation of eremomycin. Derivatives with a hydrophobic substituent at the exterior of the molecule were then synthesized, and their antibacterial activities were compared with similar derivatives of eremomycin. Comparison of derivatives of eremomycin containing the n-decyl or p-(p-chlorophenyl)benzyl substituent in the eremosamine moiety (N') and n-decyl or p-(p-chlorophenyl)benzylamides with similar derivatives of eremomycin possessing the damaged peptide core (a defective binding pocket) showed that compounds of both types are almost equally active against glycopeptide-resistant strains of enterococci (GRE), whereas eremomycin derivatives are more active against staphylococci. Hydrophobic 7d alkylaminomethylated derivatives of eremomycin (9, 10) demonstrated similar antibacterial properties. Since the basic mode of action of glycopeptide antibiotics involves binding to cell wall intermediates terminating in -D-Ala-D Ala and this interaction is seriously decreased in the hexapeptide derivatives (lacking the critical N-methyl-D-leucine), we suggest that these hydrophobic derivatives may inhibit peptidoglycan synthesis in the absence of dipeptide binding. NMR binding experiments using Ac-D-Ala-D-Ala show that binding constants of these hexapeptide derivativies are decreased in comparison with the corresponding heptapeptides with intact binding pocket. This is in agreement with the decreased biological activity of the hexapeptide derivatives against vancomycin-sensitive strains in comparison with the activity of parent compounds. Binding to the lactate cell wall analogue Ac-D-Ala-D-Lac with decylamide of eremomycin 8 was not observed, demonstrating that the interaction with this target in GRE does not occur. While hydrophobic glycopeptide derivatives retain the ability to inhibit the synthesis of peptidoglycan in manner of natural glycopeptides, biochemical investigation supports the hypothesis that they inhibit the transglycosylase stage of bacterial peptidoglycan biosynthesis even in the absence of dipeptide or depsipeptide binding. PMID- 11882004 TI - Design and synthesis of a novel and potent series of inhibitors of cytosolic phospholipase A(2) based on a 1,3-disubstituted propan-2-one skeleton. AB - Using knowledge of the substrate specificity of cPLA(2) (phospholipases A(2)), a novel series of inhibitors of this enzyme were designed based upon a three point model of inhibitor binding to the enzyme active site comprising a lipophilic anchor, an electrophilic serine "trap", and an acidic binding moiety. The resulting 1,3-diheteroatom-substituted propan-2-ones were evaluated as inhibitors of cPLA(2) in both aggregated bilayer and soluble substrate assays. Systematic variation of the lipophilic, electrophilic, and acidic groups revealed a well defined structure-activity relationship against the enzyme. Optimization of each group led to compound 22 (AR-C70484XX), which contains a decyloxy lipophilic side chain, a 1,3-diaryloxypropan-2-one moiety as a unique serine trap, and a benzoic acid as the acidic binding group. AR-C70484XX was found to be among the most potent in vitro inhibitors of cPLA(2) described to date being more than 20-fold more active against the isolated enzyme (IC(50) = 0.03 microM) than the standard cPLA(2) inhibitor, arachidonyl trifluoromethyl ketone (AACOCF(3)), and also greater than 10-fold more active than AACOCF(3) against the cellular production of arachidonic acid by HL60 cells (IC(50) = 2.8 microM). PMID- 11882005 TI - Stereochemistry at phosphorus of the reaction catalyzed by myo-inositol monophosphatase. AB - myo-Inositol monophosphatase (IMPase), the proposed target for lithium therapy for manic depression, is an important enzyme in the biosynthesis of second messengers. Earlier studies have shown that the IMPase-catalyzed hydrolysis of myo-inositol monophosphates to inorganic phosphate and myo-inositol proceeds by direct attack of water at phosphorus. However, research groups have independently proposed either an in-line displacement (with inversion of stereochemistry at phosphorus) or an adjacent attack with a pseudorotation (with retention of stereochemistry at phosphorus). Here, the elucidation of the stereochemical pathway is presented. The IMPase-catalyzed hydrolysis of D-1-S(p)-myo-inositol [(17)O]-thiophosphate in the presence of H(2)(18)O gave inorganic R(p) [(16)O,(17)O,(18)O]-thiophosphate, with inversion of configuration at phosphorus. This is only consistent with an in-line displacement, and it rules out the controversial adjacent/pseudorotation mechanism. This result will assist in the design of alternative inhibitors of IMPase. PMID- 11882007 TI - Design and synthesis of a novel L-dopa-entacapone codrug. AB - A novel codrug, in which L-Dopa and entacapone are linked via a biodegradable carbamate spacer to form a single chemical entity, was synthesized and studied kinetically. This carbamate codrug provides adequate stability [t(1/2) = 12.1 h (pH 1.2); 1.4 h (pH 5.0); 1.1 h (pH 7.4)] against chemical hydrolysis but rapidly hydrolyzes to L-Dopa and entacapone in liver homogenate (t(1/2) = 7 min; pH 7.4) at 37 degrees C. The therapeutical potential of this novel codrug is discussed. PMID- 11882006 TI - Synthesis and notable antimalarial activity of acyclic peroxides, L-(alkyldioxy) L-(methyldioxy)cyclododecanes. AB - Of several bis(alkyldioxy)alkanes and the related acyclic peroxides prepared in this study, 1,1-bis(methyldioxy)cyclododecane showed the most notable antimalarial activity particularly in vivo (almost a half of that of artemisinin). PMID- 11882008 TI - Synthesis and antimycobacterial activity of 6-arylpurines: the requirements for the N-9 substituent in active antimycobacterial purines. AB - 6-Arylpurines carrying a variety of substituents in the 9-position were prepared by Stille coupling between appropriately substituted 6-chloropurines and aryl(tributyl)tin, and the compounds were screened for antibacterial activity against Mycobacterium tuberculosis H(37)Rv. The lowest minimum inhibitory concentration value, 0.78 microg/mL, was found for 9-benzyl-2-chloro-6-(2 furyl)purine. This compound exhibited relatively low cytotoxicity, and it was active against several singly drug-resistant strains of M. tuberculosis. PMID- 11882009 TI - Lipoamino acid-based adjuvant carrier system: enhanced immunogenicity of group a streptococcal peptide epitopes. AB - Lipoamino acid-based synthetic peptides (lipid core peptides, LCP) derived from the type-specific and conserved region determinants of group A streptococci (GAS) were evaluated as potential candidate sequences in a vaccine to prevent GAS associated diseases, including rheumatic heart disease and poststreptococcal acute glomerulonephritis. The LCP peptides had significantly enhanced immunogenicity as compared with the monomeric peptide epitopes. Furthermore, the peptides incorporated into the LCP system generated epitope-specific antibodies without the use of any conventional adjuvant. PMID- 11882010 TI - Molecular control of luteal secretion of progesterone. AB - Cholesterol provided by low- or high-density lipoprotein is the precursor for biosynthesis of progesterone. Once inside the cell, cholesterol can be used for steroidogenesis or esterified with long-chain fatty acids and stored as cholesterol esters in lipid droplets. When it is needed for steroidogenesis, free cholesterol is transported to the mitochondrion via a mechanism that involves cytoskeletal elements and sterol carrier proteins. Cytochrome P450 cholesterol side chain cleavage enzyme complex converts the cholesterol to pregnenolone, which is then converted to progesterone by 3beta-hydroxysteroid dehydrogenase/delta5,delta4 isomerase in the smooth endoplasmic reticulum. Transport of cholesterol from the cytoplasm to the inner mitochondrial membrane is both the rate-limiting step in progesterone biosynthesis and the step most acutely influenced by second messengers. Steroidogenic acute regulatory protein (StAR) and peripheral-type benzodiazepine receptors (PBR) are involved in this transport. StAR may bind cholesterol in the cytosol and transport it to the mitochondrial membrane where PBR is involved in transport from the outer to the inner mitochondrial membrane. Phosphorylation of StAR by protein kinase A (PKA) stimulates cholesterol transport, whereas phosphorylation by PKC may inhibit this process. Endozepine, the natural ligand for PBR, also appears to be involved in regulation of the rate of cholesterol transport to the inner mitochondrial membrane and to play a role in the stimulatory effects of PKA on steroidogenesis. Increased concentrations of endozepine were detected in large luteal cells, and may explain the increased progesterone secretion from this type of cell. Fluorescence energy transfer procedures indicate that StAR associates with PBR in mitochondrial membranes. A model is presented for the proposed interactions of StAR, PBR and endozepine in the transport of cholesterol from the outer to the inner mitochondrial membrane. PMID- 11882012 TI - Characterization and biological roles of oviduct-specific, oestrogen-dependent glycoprotein. AB - During late follicular growth, oestrus, fertilization and early embryonic development, the oviduct, under specific hormonal control, produces fluid and contributes secretory macromolecules that optimize the microenvironment for gamete maturation and transport, fertilization and early cleavage-stage embryonic development. This review describes the state of knowledge concerning the physiological and biochemical characterization of the major oviduct secreted protein, the oviduct-specific, oestrogen-dependent glycoprotein. The identification, localization within the oviduct, binding and association with oocytes, embryos and spermatozoa, hormonal control of gene expression, cloning, gene organization, protein sequences and post-translational modifications of oviduct-specific, oestrogen-dependent glycoprotein are discussed. Identification of biological functions for this glycoprotein, its interactions with spermatozoa, oocytes and embryos and its potential as a paracrine regulator of fertilization and development are also discussed. PMID- 11882011 TI - Tumour suppressor genes in sporadic epithelial ovarian cancer. AB - Ovarian cancer is the most frequent cause of death from gynaecological malignancies in the western world, and sporadic epithelial ovarian cancer is its most predominant form. The aetiology of sporadic ovarian cancer remains unknown. Genetic studies have enabled a better understanding of the evolution of tumour progression. A major focus of research has been to identify tumour suppressor genes implicated in sporadic ovarian cancer over the past decade. Several tumour suppressor genes have been identified by strategies such as positional cloning and differential expression display. Further research is warranted to understand fully their contribution to the pathogenesis of sporadic ovarian cancer. PMID- 11882013 TI - Effect of bone morphogenetic protein 2 (BMP2) on oestradiol and inhibin A production by sheep granulosa cells, and localization of BMP receptors in the ovary by immunohistochemistry. AB - The bone morphogenetic proteins (BMPs) have been implicated in the paracrine regulation of ovarian follicular development. In this study, we investigated the expression of the BMP receptors (BMPRs) in sheep ovaries by immunohistochemistry and the effect of BMP2, a natural ligand for these receptors, on granulosa cells cultured in vitro. Ovaries from cyclic ewes were fixed, embedded in paraffin wax and cut into sections. The sections were rehydrated, submitted to microwave antigen retrieval and treated with polyclonal antibodies against BMPR1A, BMPR1B and BMPR2. Strong immunostaining for all three receptors was observed in the granulosa cell layer of follicles from the primary to late antral stages of development. Staining was also present in the oocyte, corpus luteum, ovarian surface epithelium and, to a lesser extent, the theca layer of antral follicles. For functional studies, granulosa cells were obtained from immature follicles 1-3 mm in diameter. The cells were cultured for 6 days in serum-free medium containing 1 ng oFSH-20 ml(-1) in the presence of 0, 3, 10 or 30 ng ml(-1) human recombinant BMP2. The medium was replaced every 2 days and oestradiol and inhibin A concentrations were measured in the spent medium. In the absence of BMP2, oestradiol and inhibin A production increased as the granulosa cells differentiated in vitro. The addition of the highest dose of BMP2 enhanced oestradiol production (P < 0.05) without affecting the proliferation of the cells. It is concluded that BMP receptors are present in sheep ovaries and that BMPs may have a role in the differentiation of granulosa cells by enhancing the action of FSH. PMID- 11882014 TI - Steroidogenesis in bovine granulosa cells: the effect of short-term changes in dietary intake. AB - The nutritional status of a cow is a key factor in the regulation of both follicle growth and oocyte quality. In this study, the effect of diets designed to increase circulating insulin and insulin-like growth factor I (IGF-I) concentrations on steroid production by granulosa cells in vitro was examined to analyse the mechanisms through which these changes occur. Hereford x Friesian heifers (n = 24) were offered maintenance or twice maintenance diets during the experimental period (17 days). Circulating concentrations of FSH did not differ between the two dietary groups, whereas insulin and IGF-I concentrations showed significant diet x day of oestrous cycle interactions. Ovaries were collected on day 3 of the first follicle wave after synchronization of oestrus. Granulosa cells were isolated from small (1-4 mm) and medium-sized (4-8 mm) follicles and cultured in the presence of long R3-IGF-I or bFSH or both. After 4 days in culture, granulosa cells isolated from small follicles, but not medium-sized follicles, collected from cattle offered the twice maintenance diet secreted significantly higher (P < 0.05) amounts of oestradiol compared with granulosa cells collected from cattle offered the maintenance diet. The effect was apparent in either the presence or absence of FSH and long R3-IGF-I. This nutritional effect on aromatase activity in granulosa cells was not apparent after day 6 of culture. There was no effect of diet on progesterone production by granulosa cells after 4 or 6 days of culture. These results support the hypothesis that dietary-induced changes in circulating insulin and IGF-I concentrations have a direct effect on the steroidogenic potential of bovine granulosa cells from small follicles. The dietary-induced increases in aromatase activity in small follicles combined with the increased concentration of metabolic hormones are possible mechanisms through which short-term changes in nutrition may affect follicle dynamics. PMID- 11882015 TI - Expression of vascular endothelial growth factor (VEGF) and its receptors in human endometrium throughout the menstrual cycle and in early pregnancy. AB - Immunohistochemistry for vascular endothelial growth factor (VEGF) and its receptors, fms-like tyrosine kinase (flt-1) and kinase insert domain-containing region (KDR), was performed on human endometrium obtained from patients with normal menstrual cycles, patients given oestrogen and progesterone, and women in early pregnancy. Intense immunostaining of VEGF was observed in both glandular epithelial and stromal cells during the mid-secretory phase; the immunostaining intensity was increased by administration of oestrogen plus progesterone and strong immunostaining was observed in decidual cells of early pregnancy. In addition to the immunostaining in vascular endothelial cells, strong KDR immunostaining was observed in glandular epithelial cells and in decidualized stromal cells induced by administration of oestrogen plus progesterone, whereas flt-1 immunostaining was negligible. Strong immunostaining for flt-1 and KDR was found in both vascular endothelial cells and decidual cells in early pregnancy. Endometrial stromal cells isolated from proliferative phase endometrium were incubated with oestrogen (10(-8) mol l-1) and medroxyprogesterone acetate (MPA; 10(-6) mol l-1) for 18 days to study the regulation of VEGF, flt-1 and KDR in endometrial stromal cells by oestrogen and progesterone. Expression of VEGF and KDR mRNAs was increased significantly by oestrogen and MPA, accompanied by decidualization, whereas flt-1 mRNA expression was not affected. In conclusion, VEGF and its receptors may play important roles in implantation and maintenance of pregnancy. PMID- 11882016 TI - Expression of a blood-brain barrier-specific antigen in the reproductive tract of the male rat. AB - The endothelial barrier antigen (EBA) is a protein expressed specifically by the endothelial cells of the rat brain barrier vessels. This antigen has been described as a 'barrier protein' and is used as a marker for the competent blood brain barrier. A blood-testis barrier has also been described. However, unlike the blood-brain barrier, which is formed by endothelial cells, the blood-testis barrier is formed mainly by the Sertoli cells, which provide an isolated environment for spermatogenic cells within the seminiferous tubules. Testicular blood vessels express the erythroid glucose transporter protein and other markers, which are strongly expressed in brain blood vessels, and may contribute to the blood-testis barrier. This study was carried out to determine whether Sertoli cells or testicular blood vessels express EBA. Tissues of other organs were used as controls for EBA expression. EBA was expressed by the endothelial cells in most microvessels of the testis, and in a few vessels of the epididymis, seminal vesicle, prostate gland, vas deferens and bladder-neck region. Furthermore, EBA was strongly and consistently detected in epithelial cells of the rete testis and dorsolateral prostate gland, and in a few epithelial cells of the ventral prostate gland, the seminal vesicle and the coagulating gland. However, Sertoli cells, which are the main site of the blood-testis barrier, were negative for EBA. In conclusion, EBA may have a wider role in rat tissues than has been previously appreciated. PMID- 11882017 TI - Cytokine expression in the uterus of mice with pregnancy loss: effect of maternal immunopotentiation with GM-CSF. AB - It is believed that failure of the maternal immune system to actively support embryonic development, through production of the appropriate cytokine network, might be responsible for embryonic death. Thus, the aim of this study was to evaluate the possible involvement of cytokines such as tumour necrosis factor alpha (TNF-alpha) and transforming growth factor beta2 (TGF-beta2), which are crucial for normal embryonic development, in the early stages of mechanisms that mediate induced pregnancy loss. The early stages of the resorption process induced by lipopolysaccharide (LPS) were characterized by blood accumulation in the vicinity of the embryo, preceding any visible embryonic damage. At that time, immunohistochemical analysis revealed an increased expression of TNF-alpha in the primary and secondary decidua, which was reduced as the resorption process was completed. In contrast, TGF-beta2 expression was decreased in the primary and secondary decidua, as well as in the glandular epithelium, at all the times assessed. Maternal immunopotentiation with granulocyte macrophage-colony stimulating factor (GM-CSF), which controls maternal immune activities supporting normal embryonic development, decreased the resorption rate in LPS-treated mice while normalizing the expression of TNF-alpha and TGF-beta2 in the uterus of these animals throughout the ongoing resorption process. These results indicate a possible role for maternal immunopotentiation with GM-CSF in the mechanisms mediating the early stages of pregnancy loss, possibly via modulation of TNF alpha and TGF-beta2 activity. PMID- 11882018 TI - Abnormal expression of acid glycosidases in seminal plasma and spermatozoa from infertile men with varicocele. AB - The activities of acid beta-glucuronidase, alpha-mannosidase, alpha-glucosidase, alpha-galactosidase, beta-galactosidase and beta-N-acetylglucosaminidase were analysed in seminal plasma and spermatozoa from 26 infertile men with varicocele and from 36 men of normal fertility. Semen samples from ten men with non obstructive azoospermia were used as control specimens that contained the other components of semen. Spermatozoa were solubilized by both physical (homogenization) and chemical (Triton-X100) methods to obtain the soluble and non soluble fractions. The activities of several glycosidases measured both in seminal plasma and spermatozoa were directly correlated with the numbers of spermatozoa and sperm motility, confirming previous studies. As some infertile patients with varicocele have normal semen parameters, whereas others have low numbers of spermatozoa and low sperm motility, the varicocele patients were prospectively divided into two groups: one (n = 15) with normal spermiograms and the other (n = 11) with abnormal spermiograms. The activities (expressed in mU ml(-1)) of alpha-mannosidase, beta-galactosidase and beta-N-acetylglucosaminidase in seminal plasma of normozoospermic infertile patients with varicocele were significantly higher than those of fertile controls, but not when expressed in U per 10(8) spermatozoa. The activities of beta-glucuronidase, alpha-mannosidase, beta-galactosidase and beta-N-acetylglucosaminidase in seminal plasma when expressed in U per 10(8) spermatozoa in varicocele patients with abnormal spermiograms were significantly higher than in those of men of normal fertility. The activity of alpha-mannosidase in the soluble fraction of sperm homogenates, expressed as U per 10(8) spermatozoa, was significantly higher in infertile patients with varicocele and abnormal spermiograms than in controls. In the non soluble fraction of spermatozoa from infertile patients with varicocele, there was an increase in the expression of beta-galactosidase and beta-N acetylglucosaminidase activities compared with the fraction of spermatozoa from fertile subjects. In summary, infertile patients with varicocele displayed an overexpression of acid alpha-mannosidase, beta-galactosidase and beta-N acetylglucosaminidase activities in seminal plasma and spermatozoa that may be associated with functional defects in spermatozoa as these glycosidases play an important role in mammalian fertilization. PMID- 11882019 TI - Effect of dietary-induced increases in circulating insulin concentrations during the early postpartum period on reproductive function in dairy cows. AB - Although it has become increasingly clear that fertility in modern dairy cattle is declining in association with increased milk yields, the underlying mechanism is poorly understood. The first ovulation post partum is delayed in dairy cows undergoing selection for genetic merit for milk yield in association with lower circulating insulin concentrations. The aim of this study was to investigate whether feeding a diet to increase circulating insulin concentrations can overcome this delay in the first ovulation post partum. The experiment was a 2 x 2 factorial design (n = 10 per group) involving diet and genetic merit for milk yield. The dietary treatment started on the day of calving and lasted for 50 days. Plasma samples were collected each day and ovarian ultra-sonography was performed three times a week during the experimental feeding period. Milk yield was recorded each day, and body weight and body condition score were determined each week. Milk samples were collected three times a week from day 50 to day 105 post partum, and reproductive performance data were recorded for all the cows as part of the routine farm practice. The dietary treatment induced significant differences in plasma insulin concentrations in both high and low genetic merit cows. Although high genetic merit cows produced more milk, lost more body weight and had lower body condition scores during the experiment, no significant effect of diet was observed on these measurements. The high insulin inducing diet increased the proportion of cows ovulating within 50 days of calving and reduced the intervals from calving to first ovulation, and tended to reduce the intervals from calving to first service and to conception. These fertility parameters were also more favourable in low than in high genetic merit cows, but no interaction between diet and genetic merit was observed for any of these parameters. Genetic merit, but not diet, also affected the number of services required per conception and the conception rate. In conclusion, these results have confirmed that genetic selection for high milk yield is associated with a decrease in reproductive performance in dairy cows. More importantly, this study has demonstrated that it is possible to alleviate this problem by nutritional manipulation. PMID- 11882020 TI - Role of PGF2alpha and oxytocin in parturition in the brushtail possum (Trichosurus vulpecula). AB - Maturation of the fetal pituitary and adrenal glands allows the secretion of cortisol, which in turn leads to an increase in prostaglandin and mesotocin production. The production of prostaglandin and mesotocin results in an increase in uterine contractions and initiates birth in marsupials. The major metabolite of PGF(2alpha), 13,14-dihydro-15-keto-prostaglandin F(2alpha) (PGFM), has been found in the plasma of the possum at the time of birth and administration of PGF(2alpha) to female possums induced the adoption of the birth position. Evidence that mesotocin is an integral hormone of birth in the tammar wallaby indicates that both PGF(2alpha) and mesotocin or oxytocin are required for marsupial birth. The presence of PGF(2alpha) receptors in the uterus and corpus luteum of the possum, and the in vitro uterine responsiveness to PGF(2alpha) or oxytocin, were examined. PGF(2alpha) receptors were not observed in possum uteri and the inability of PGF(2alpha) to cause contractions indicates that PGF(2alpha) is not involved directly in contraction of the uterus at parturition. The presence of oxytocin and mesotocin receptors in the uterus of possoms and the ability of oxytocin to induce uterine contraction in vitro supports the view that mesotocin is required for expulsion of the young from the uterus. Low numbers of PGF(2alpha) receptors were found in the possum corpus luteum at birth, indicating an involvement of PGF(2alpha) in regression of the corpus luteum. PMID- 11882021 TI - Behaviour of a sperm surface transmembrane glycoprotein basigin during epididymal maturation and its role in fertilization in mice. AB - Basigin (bsg) is a transmembrane glycoprotein belonging to an immunoglobulin superfamily and is localized on the surface of the sperm tail. The behaviour of bsg during epididymal maturation and its role in fertilization were examined using an anti-bsg antibody. Spermatozoa from caput, corpus and cauda epididymides were immunostained by indirect immunofluorescence (IIF). Immunostaining revealed that bsg is localized on the principal piece of caput spermatozoa and the molecule was found on the middle piece during transit in the corpus and cauda epididymides. Concomitantly, the molecular mass of bsg was reduced from 37 kDa (testis) to 26 kDa (cauda epididymidis). IVF experiments were designed to assess the effect of anti-bsg antibody on the fertilization events. Anti-bsg antibody significantly inhibited primary binding to the cumulus-invested oocytes with intact zonae pellucidae in a dose-dependent manner. Consequently, the fertilization rate of cumulus-invested oocytes with intact zonae pellucidae was also inhibited. The bsg molecule was also detected on the head of live capacitated spermatozoa by IIF under IVF conditions. These findings indicate that testicular bsg is a glycosylated protein that undergoes molecular processing and deglycosylation during its transit in the epididymis. The bsg molecule that was detected on the sperm head after capacitation may facilitate the primary binding or might be involved in distinct events required for primary binding of spermatozoa to the zona pellucida during capacitation and sperm-cumulus interaction. PMID- 11882022 TI - Influence of maternal size on placental, fetal and postnatal growth in the horse. I. Development in utero. AB - The interacting influences of maternal size and fetal genotype on placental and fetal development in the mare were assessed by comparing conventional within breed Thoroughbred (Tb-in-Tb, n = 7) and Pony (P-in-P, n = 7) control pregnancies established by artificial insemination (AI) with between-breed (Tb-in-P, n = 8; deprived in utero condition and P-in-Tb, n = 7; luxurious in utero condition) experimental pregnancies established by embryo transfer. All foals were born spontaneously and the mean (+/- SEM) duration of gestation in the two groups of control mares was significantly different (P < 0.001) at 325 +/- 3.0 days for the P-in-P pregnancies and 339 +/- 3.0 days for the Tb-in-Tb pregnancies, whereas the durations of gestation for the two experimental groups were very similar and midway between those of the control pregnancies at 332 +/- 2.8 days for the Tb-in P and 331 +/- 2.7 days for the P-in-Tb. Mean (+/- SEM) foal birth weight and the mean (+/- SEM) values for the mass, gross area and volume of the allantochorion were all highest in the seven Tb-in-Tb pregnancies (53.1 +/- 2.6 kg, 3.8 +/- 0.3 kg, 12.9 +/- 0.3 x 10(3) cm(2), 3.5 +/- 0.2 l, respectively) and lowest in the seven P-in-P control pregnancies (24.0 +/- 1.3 kg, 1.7 +/- 0.1 kg, 8.3 +/- 0.3 x 10(3) cm(2), 1.8 +/- 0.1 l, respectively). These parameters were higher in the seven P-in-Tb pregnancies (37.9 +/- 2.1 kg, 2.7 +/- 0.1 kg, 10.1 +/- 0.5 x 10(3) cm(2), 2.5 +/- 0.1 l, respectively) than in the eight Tb-in-P (33.0 +/- 2.4 kg, 2.3 +/- 0.2 kg, 9.0 +/- 0.5 x 10(3) cm(2), 2.1 +/- 0.1 l) experimental pregnancies. Foal birth weight was positively correlated with the mass (r = 0.84, P < 0.001), gross area (r = 0.87, P < 0.001) and volume (r = 0.91, P < 0.001) of the allantochorion, and maternal weight was also positively correlated with both the mass and gross area of the allantochorion (r = 0.64 and 0.69, respectively; both P < 0.001). Application of stereology to multiple random biopsies recovered from each placenta produced mean values for the surface density of microcotyledons on the allantochorion (S(v)). Values were higher in Thoroughbred than in Pony mares regardless of the breed of fetus being carried. Multiplication of S(v) by the volume of the allantochorion to give the total microscopic area of fetomaternal contact at the placental interface was also positively correlated with foal birth weight (r = 0.84, P < 0.001). Foal birth weight was determined by the microscopic area of fetomaternal contact of the placenta and there were no differences in foal weight per m(2) of placenta regardless of fetal or maternal genomes. Thus, the results indicate that in equids, maternal size interacts with both the maternal and fetal genotypes to control the rate and extent of fetal growth by influencing the gross area of the diffuse allantochorion, and the density, complexity and depth of the microcotyledons on its surface. PMID- 11882023 TI - Developmental competence in vivo and in vitro of in vitro-matured equine oocytes fertilized by intracytoplasmic sperm injection with fresh or frozen-thawed spermatozoa. AB - This study was undertaken to evaluate the development of equine oocytes in vitro and in vivo after intracytoplasmic sperm injection (ICSI) with either fresh or frozen-thawed spermatozoa, without the use of additional activation treatments. Oocytes were collected from ovaries obtained from an abattoir and oocytes classified as having expanded cumulus cells were matured in M199 with 10% fetal bovine serum and 5 microU FSH ml(-1). After 24-26 h of in vitro maturation, oocytes with a first polar body were selected for manipulation. Fresh ejaculated stallion spermatozoa were used for the experiment after swim-up for 20 min in sperm-Tyrode's albumen lactate pyruvate. Frozen-thawed spermatozoa from the same stallion were treated in a similar way. Spermatozoa were immobilized and injected into the oocytes using a Piezo drill. Presumptive zygotes were cultured in G1.2 medium for 20 or 96 h after the injection was administered, or were transferred to the oviducts of recipient mares and recovered 96 h later. In addition, bovine oocytes with first polar bodies were injected with the two types of stallion spermatozoa and fixed 20 h after injection to examine pronuclear formation. Fertilization rate (pronucleus formation and cleavage) at 20 h after injection of spermatozoa was not significantly different between fresh and frozen-thawed sperm groups in either equine or bovine oocytes. Pronucleus formation after injection of spermatozoa into bovine oocytes was significantly higher than that for equine oocytes (P < 0.05). There were no significant differences in cleavage rate or average number of nuclei at 96 h between equine oocytes injected with fresh or frozen-thawed spermatozoa. However, embryos developed in vivo for 96 h had a significantly higher number of nuclei in both sperm treatments compared with those cultured in vitro. These results indicate that good activation rates may be obtained after injection of either fresh or frozen-thawed equine spermatozoa without additional activation treatment. Injection of frozen-thawed equine spermatozoa results in similar embryo development to that obtained with fresh equine spermatozoa. In vitro culture of equine zygotes in G1.2 medium results in a similar cleavage rate but reduced number of cells compared with in vivo culture within the oviduct. Bovine oocytes may be useful as models for assessing sperm function in horses. PMID- 11882024 TI - Matrix metalloproteinases (MMPs) in the endometrium of bitches. AB - The relationships between activities of matrix metalloproteinases (MMPs) in the canine uterus and the occurrence of degeneration of the luminal epithelium, cystic endometrial hyperplasia, pyometra and uterine remodelling post partum were determined. Mature bitches (n = 10) were ovariectomized, treated with hormones (oestradiol benzoate, progestagen) and investigated at stages simulating pro oestrus (n = 2), oestrus (n = 2), dioestrus (n = 2), and mid- (n = 2) and late (n = 2) anoestrus (3 and 9 weeks, respectively, after cessation of treatment with progestagen). Untreated bitches (n = 1 per group) served as controls (Expt 1). An additional 10 ovariectomized bitches, at the end of treatment-induced simulated dioestrus, were treated each day for a further 3 weeks either with the same dose (standard dose, n = 3), a decreased dose (n = 3) or an increased dose (n = 3) of progestagen, or no treatment (withdrawal dose, n = 1). These bitches were then investigated (Expt 2). A suture was placed in the lumen of one uterine horn of another five bitches at ovariectomy. Three of these bitches were treated to induce simulated dioestrus and two bitches served as untreated controls. In the hormone-treated bitches, the suture resulted in cystic endometrial hyperplasia in one bitch and in cystic endometrial hyperplasia with pyometra in two bitches. The control bitches showed no cystic endometrial hyperplasia or pyometra (Expt 3). Four intact bitches were studied at 2 (n = 1), 3 (n = 2) and 11 (n = 1) weeks post partum. Uterine tissues were also collected from two bitches with naturally occurring cystic endometrial hyperplasia with pyometra (Expt 4). All uteri were examined histologically and the activities of MMP-2, -7 and -9 (latent and active forms) were assessed using zymography of extracts of endometrium. In Expts 1 and 2, marked degeneration of the luminal epithelium in six of 25 bitches (simulated mid-anoestrus, withdrawal dose and decreased dose groups) was not associated with changes in MMP activities. Markedly increased activities of MMP-2 (active form), 7 (latent form) and -9 (active and latent forms) were observed in the bitches with cystic endometrial hyperplasia with pyometra (but not with cystic endometrial hyperplasia alone) and in the bitches at 2 and 3 weeks post partum (but not at 11 weeks post partum). These results indicate that MMPs are not involved with degeneration of the luminal epithelium, but are involved with uterine remodelling in the postpartum bitch and with cystic endometrial hyperplasia with pyometra. PMID- 11882025 TI - Heat shock proteins and innate immunity. PMID- 11882026 TI - Transgenic models of autoimmune disease. AB - Transgenic and knockout mouse models have been invaluable for the elucidation of basic mechanisms in autoimmunity and have contributed new experimental models of human autoimmune diseases. Transgenic models of self tolerance have helped to change our view of this state from a process mediated purely by thymic deletion to a more complex process encompassing deletion, peripheral anergy, down regulation of receptors and modulation by regulatory cells. Experiments in which the genes for the candidate target antigens in autoimmune disease are over expressed or under-expressed have helped to clarify the targets of attack. Several examples of T cell receptor transgenic mice have been described in which T cells carry the receptor derived from a human or mouse autoimmune T cell clone. Such mice allow the characterization of T cell specificities contributing to disease and of the additional factors and checkpoints influencing disease development. In addition, the expression of disease associated HLA alleles in 'humanised' transgenic lines allows the mapping of HLA-restricted T cell epitopes and investigation of the mechanisms underlying these genetic associations. These approaches are leading to the generation of new disease models, offering hope for the design and testing of novel immunotherapeutic strategies. PMID- 11882027 TI - B7-1 and B7-2 co-stimulatory molecules are required for mercury-induced autoimmunity. AB - B7-1 (CD80) and B7-2 (CD86) molecules on antigen presenting cells play important roles in providing co-stimulatory signals required for activation and expansion of autoreactive T cells. Moreover, some reports have suggested that these molecules may have distinct functions in the differentiation of Th1 and Th2 cells. Mercury-induced autoimmunity in H-2s mice is characterized by lymphoproliferation of T and B cells, serum increases in IgG1 and IgE and production of antinucleolar antibodies (ANoA). The mechanisms responsible for the various manifestations of this syndrome have yet to be elucidated. To examine the contributions of B7 co-stimulatory molecules to this model, susceptible mice were treated with antibodies to B7-1, B7-2, or both during the development of mercury induced autoimmunity. The combination of anti-B7-1 and anti-B7-2 antibodies prevented Hg-induced disease in H-2s mice. Additionally, single anti-B7-1 antibody treatment was sufficient to prevent Hg-induced ANoA production, but not IgG1 and IgE hypergammaglobulinaemia. Further, single antibody treatment with anti-B7-2 resulted in a partial reduction of ANoA titres but had no significant effect on total serum IgG1 and IgE levels. Taken together, these results indicate that B7-1 and B7-2 molecules are critical for the development of Hg-induced autoimmunity and suggest that the different manifestations of the syndrome are regulated by independent mechanisms. PMID- 11882028 TI - IFN-gamma and IL-12 but not IL-10 are required for local tumour surveillance in a syngeneic model of orthotopic bladder cancer. AB - In recent studies, a crucial role for IFN-gamma in immunosurveillance of tumours and in IL-12 immunotherapy has been suggested. Nevertheless, little is known about the relevance of IFN-gamma and IL-12 for tumour surveillance in noncytokine immunotherapy. Adjuvant immunotherapy with viable BCG (Bacillus Calmette--Guerin) is considered to be the most powerful clinical treatment regimen of bladder cancer and is known to induce a variety of proinflammatory cytokines. Consequently, we analysed the antitumour response of IFN-gamma knockout (KO), IL 12 KO and IL-10 KO mice in the absence and presence of BCG immunotherapy in a syngeneic orthotopic model of bladder cancer. IFN-gamma KO and IL-12 KO mice died much earlier and by far smaller tumour inocula compared to wildtype mice, while this intrinsic antitumour response was not altered in IL-10 KO mice. BCG immunotherapy was effective in wildtype mice, but totally ineffective in IFN gamma KO and IL-12 KO mice. BCG induced a massive local immune response in the bladder of treated animals. This response was markedly increased in IL-10 KO mice, which coincides with increased therapeutic efficacy in this mouse strain compared with wildtype mice. Our data establish a crucial role for a Th1 type immune response in the intrinsic and immunotherapeutic control of local orthotopic bladder cancer. PMID- 11882029 TI - Trypanosoma cruzi mitochondrial malate dehydrogenase triggers polyclonal B-cell activation. AB - It has been proposed that Trypanosoma cruzi, the aetiologic agent of Chagas' disease, produces mitogenic substances responsible for the polyclonal B-cell activation observed during the acute phase of the infection. Isolation and characterization of the molecules involved in the induction of polyclonal activation observed during infectious diseases have posed a great challenge for the immunologist over the last decade. In this work we report that a 33 kD protein obtained from an alkaline fraction of T. cruzi epimastigotes (FI) stimulates proliferation and promotes differentiation into antibody-secreting cells of normal murine B cells in a T-cell independent manner. By flow cytometry we also found that the 33 kDa protein induces an increase in the expression of MHC class II and B7.2 but not B7.1 molecules on the B-cell surface. Sequencing by mass spectrometry identified the T. cruzi 33 kD protein as hypothetical oxidoreductase, a member of the aldo/ketoreductase family. In this report we demonstrate that this protein is also present in the infective bloodstream trypomastigote form of the parasite and was identified as T. cruzi mitochondrial malate dehydrogenase (mMDH) by enzyme activity and by Western blotting using a specific mMDH polyclonal antiserum. The biologic relevance of mMDH-induced polyclonal activation concerning T. cruzi infection is discussed. PMID- 11882030 TI - Extra-articular cartilage affected in collagen-induced, but not pristane-induced, arthritis models. AB - Rheumatoid arthritis (RA) is a chronic inflammatory disease primarily affecting cartilaginous joints but also extra-articular tissues such as the nose and upper respiratory tract. We have investigated extra-articular cartilage involvement in two commonly used animal models for RA, collagen-induced and pristane-induced arthritis, by immunizing rats with different susceptibility to disease (LEW.1 A, LEW.1F and DA rats). We found that nasal and tracheolaryngeal cartilage is affected in LEW.1 A and DA rats to varying degrees in collagen-induced arthritis but not in any strain in the pristane-induced model. Antibodies to matrilin-1, a cartilage-specific protein expressed mainly in tracheolaryngeal and nasal cartilage but not in joints, were positively associated with the presence of inflammation in nasal cartilage. In contrast, no antibody response to matrilin-1 could be detected in pristane-induced arthritis. In addition, nasal vaccination with collagen type II prior to immunization in DA rats significantly decreased the antibody response to matrilin-1 at day 56, but not at earlier time points, indicating a late protective effect on extra-articular cartilage. We conclude that pristane-induced arthritis is a joint-specific model whereas collagen induced arthritis affect joints as well as extra-articular cartilage. Furthermore, collagen immunization induces an antibody response to matrilin-1. PMID- 11882031 TI - Thymic re-entry of mature activated T cells and increased negative selection in vascularized allograft recipients. AB - Transplantation tolerance is a dynamic state that involves several homeostatic mechanisms intrinsic to the host. One of these mechanisms is activation-induced T cell death (AICD). However, it is unclear where AICD takes place during alloreactive responses. Since activated T cells can re-enter the thymus, we hypothesized that mature T cells activated by an allograft could be deleted upon re-entry into the thymus. To test this hypothesis, we used wild-type or 2C TCR transgenic mice receiving syngeneic or allogeneic heterotopic, vascularized heart grafts. First, we demonstrated that ex vivo CFSE-labelled T cells re-entered the thymus when transferred into allograft recipients but not when transferred into isograft recipients. Next, we compared the changes in cell subset numbers and incidence of apoptosis in the thymi and spleens of allograft or isograft recipients. Seven days after transplantation, at a time in which all the allografts were undergoing rejection, cells expressing donor-MHC class II molecules had migrated to the thymus and to the spleen. In the thymus of allograft recipients, overall cellularity was significantly reduced by 40% and associated with an increase in the number of double negative (CD4-CD8-) thymocytes and a decrease in double positive (CD4+CD8+) thymocytes, consistent with increased negative selection of thymocytes. Additionally, thymi of allograft recipients showed an increase in the number of recently activated, mature T cells (TCRhi, CD25+, CD44+) and a significant increase in the number of apoptotic cells, especially in the thymic medulla, that involved mature T cells as indicated by the TCRhi, CD44+, CD4 or CD8 single positive phenotype. Spleens of allograft recipients were increased in size and cellularity but did not show any of the changes in cell subsets seen in the thymi. Our data show that after allografting there is an increase in apoptotic cell death that is associated with negative selection of developing thymocytes as well as of alloreactive mature T cells that have re-entered the thymus upon activation in the periphery. This may occur upon migration of graft-derived antigen-presenting cells to the thymus. PMID- 11882032 TI - IL-1ra and IL-1 production in human oral mucosal epithelial cells in culture: differential modulation by TGF-beta1 and IL-4. AB - Regulatory cytokines mediate the participation of oral mucosal epithelial cells (OMEC) in local immune responses. The aim of this study was to characterize the isoforms of IL-1 receptor antagonist (IL-1ra) in cultured human primary OMECs and to compare its production with that of IL-1 alpha (IL-1alpha) and IL-1 beta (IL 1beta). Western blot analysis showed that IL-1ra was 22 kDa in size hence slightly smaller than monocyte IL-1ra (25 kDa). A minor form of 20 kDa was also found in unstimulated cell culture lysates. In culture supernatants, IL-1 bioactivity increased after IL-1ra neutralization, indicating that the baseline production of IL-1ra is biologically relevant. Immunohistochemistry showed a relation between IL-1ra and involucrin expressions, suggesting that intracytoplasmic IL-1ra may be involved in cell terminal differentiation. In unstimulated culture lysates, there was far more IL-1ra than IL-1alpha and IL 1beta. TGF-beta1 markedly increased the IL-1ra/IL-1beta ratio from 93.6 : 1 to 300 : 1. IL-4, which is generally described as an anti-inflammatory cytokine, increased IL-1 but not IL-1ra production. TNF-alpha increased intracellular production of the three IL-1 members. IL-1ra levels were lower in supernatants than in lysates of cultured cells. Our results show that human OMECs constitutively produce significant amounts of a biologically active form of IL 1ra. TGF-beta1 mu(p)-regulation points to a positive amplification loop and IL-4 to a down-regulation loop, both including Th2 cells and OMECs. They may be important in oral tolerance and IgA production, respectively. PMID- 11882033 TI - Differential expression of beta1 and beta2 integrins and L-selectin on CD4+ and CD8+ T lymphocytes in human blood: comparative analysis between isolated cells, whole blood samples and cryopreserved preparations. AB - Flow cytometric analysis was used to compare the expression of adhesion molecules on human CD4+ and CD8+ T lymphocytes in isolated blood mononuclear cells (MNCs) in whole blood samples and in cryopreserved MNC preparations. Examination of MNCs revealed that the CD11b and CD11c components of the beta2 integrins were preferentially expressed on CD8+ T cells, whereas CD62L was present on more CD4+ T cells. All CD4+ and CD8+ T lymphocytes were positive for CD11a but the CD8+ population had a higher intensity of expression of CD11a and also CD11b. Virtually identical results were obtained with T cells in whole blood samples. In relation to the beta1 integrins, the only difference between isolated CD4+ and CD8+ T cells was that the latter subset had a greater proportion of cells bearing CD49d. The naive cell marker CD45RA was present on the majority of CD8+ T cells whereas CD45RA and the memory marker CD45RO were evenly distributed within the CD4+ T cell subset. Although cryopreservation of lymphocytes did not modify the expression of beta1 and beta2 integrins it produced a marked reduction in the percentage of CD4+ and CD8+ T cells bearing CD62L. With regard to endothelial interactions, it appears that cryopreserved lymphocytes are suitable for inclusion in studies of integrin-mediated adhesion but not for those relating to tethering or recognition of addressins on high endothelial venules. Differences in adhesion molecule expression between CD4+ and CD8+ T lymphocytes could underlie the selective extravasation of these subsets into sites of infection and inflammation. PMID- 11882034 TI - Regulation of primary HIV-1 isolate replication in dendritic cells. AB - The potential role of dendritic cells (DC) in the immunopathology of human immunodeficiency virus 1 (HIV-1) disease remains controversial. This study examines replication of a panel of HIV-1 strains (both laboratory adapted and primary) within DC, in the context of the well-established monocyte-DC and monocyte-macrophage transition. Viral replication was assessed by p24 ELISA assay. All strains of HIV-1 tested replicated in DC. Only CCR5-tropic virus replicated in macrophages. Lipopolysaccharide (LPS) induced DC maturation (as reflected in altered cell phenotype) and at the same time diminished the ability of DC to support HIV-1 replication. In contrast the presence of activated T cells, which had been fixed to prevent them acting as a site for viral replication, enhanced the ability of the DC to support viral replication, as has been reported previously for macrophages. Thus cells that are DC by phenotype, but are not activated, act as the optimum reservoir for HIV-1 replication. If this form of DC is present in peripheral tissues, this will be permissive for amplification of the in vivo viral load at sites where there are few responder cells available, and hence contribute to the persistent immunopathology. PMID- 11882035 TI - Self-heat shock protein 60 induces tumour necrosis factor-alpha in monocyte derived macrophage: possible role in chronic inflammatory periodontal disease. AB - Heat shock protein 60 (hsp60) has been increasingly recognized as an important molecule in infectious and autoimmune diseases. We have demonstrated previously that serum antibodies to both human hsp60 and Porphyromonas gingivalis GroEL were elevated in periodontitis patients compared with healthy subjects. In order to clarify the relative importance of hsp60 in the inflammatory response in periodontal disease, the stimulatory effect of human and bacterial hsp60 on the production of tumour necrosis factor-alpha (TNF-alpha) was examined in phorbol myristate acetate (PMA)-stimulated THP-1 cells. As bacterial hsp60s, recombinant P. gingivalis and Actinobacillus actinomycetemcomitans GroEL was used. Human hsp60 but not P. gingivalis or A. actinomycetemcomitans GroEL demonstrated stimulatory activity similar to lipopolysaccharide (LPS) derived from the bacteria. The activity of hsp60 was inhibited by anti-CD14 and anti-Toll-like receptor 4 (TLR4) antibodies, suggesting that both CD14 and TLR4 mediate hsp60 signalling. Immunohistochemical analysis demonstrated that hsp60 is abundantly expressed in periodontitis lesions. Therefore, it is postulated that periodontopathic bacteria stimulate the cells in the periodontium to up-regulate the expression of hsp60, which in turn may stimulate macrophage and possibly other cells to produce proinflammatory cytokines. These mechanisms may be involved in the chronicity and tissue destruction of periodontal disease. PMID- 11882036 TI - The effect of IL-18 on IL-12-induced CD30 expression and IL-4 and IFN-gamma production by allergen and PPD specific T cells. AB - CD30 is expressed on activated T cells that, as has been suggested, preferentially produce IFN-gamma. Interleukin 12 increases antigen-induced CD30 expression on T cells and IFN-gamma production. Synthesis of IFN-gamma can be augmented further by IL-18. The aim of our study was to investigate whether IL-18 affects the IL-12 induced CD30 expression and cytokine production by allergen or PPD specific T cells. Mononuclear cells of healthy or atopic volunteers were stimulated with PPD or allergen, respectively, to obtain specific T cell lines. T cells were restimulated with appropriate antigen and antigen-presenting cells in the presence of IL-12, IL-18 or a combination of these cytokines. After 3 days, expression of CD30 was investigated on CD4 and CD8 T cells and IFN-gamma and IL-4 cytokine production was estimated in the culture supernatants. Flow cytometric analyses showed no effect of IL-18 on CD30 expression during IL-12 co stimulation. At the same time after the optimal stimulation for CD30 expression, the levels of IFN-gamma were high in PPD-stimulated cell lines but have not been up-regulated by IL-18. IFN-gamma levels were much lower in allergen-stimulated T cells and although they were up-regulated by IL-12 there was no additional or synergistic effect from IL-18. IL-18, however, increased production of IL-4 in allergen-stimulated cell lines. Our studies provide new information about IL-18 activity on human cells and question its exclusive role in Th1 mediated responses. PMID- 11882037 TI - Proliferation of T-cell subsets that contact tumour cells in colorectal cancer. AB - We have investigated the proliferation rates of T-cell subsets in colorectal carcinomas using immunohistochemistry. It was found that the tumour-infiltrating T cells in contact with the tumour cells have a significantly higher frequency of proliferation than those in the stroma. In particular, the CD8+ intraepithelial lymphocytes (T-IEL) within the tumours have a significantly higher frequency of proliferation in comparison with CD8+ T cells in the stromal compartment or in any normal mucosal lymphoid tissues. It is possible that the proliferation of the CD8+ T-IEL may be driven by self-antigens expressed on the tumour cells. The proportion of CD3+ CD7- T cells is increased within carcinomas compared with the normal colon, and a population of CD57+ T cells was observed which is absent from the normal colon. It is possible that these phenotypes are acquired in situ due to repeated stimulation of the T cells by tumour antigens. Intact colorectal carcinoma explants were cultured, and the presence of tumour-infiltrating T cells analysed after 3 days of culture in isolation from the systemic compartments. CD3+ T cells were proliferating (at a low rate) within the explants after 3 days of culture, indicating that they may be sustained by factors present in the tumour microenvironment. PMID- 11882038 TI - Drug-induced neutropenia associated with anti-neutrophil cytoplasmic antibodies (ANCA): possible involvement of complement in granulocyte cytotoxicity. AB - Although antineutrophil antibodies are thought to be involved in drug-induced neutropenia, neither the precise mechanisms nor the particular antigens on the neutrophil surface have yet been clarified. Recently, we examined a patient with Graves' disease who developed antineutrophil cytoplasmic antibodies (ANCA) after propylthiouracil treatment and exhibited neutropenia. Because several target antigens of ANCA are expressed on the surface of neutrophils, it was suggested that ANCA might contribute to neutropenia. The patient's serum bound specifically to neutrophils and HL-60 cells differentiated into granulocytes, and lysed the HL 60 cells via a complement-mediated mechanism. Furthermore, two representative ANCA antigens, proteinase 3 and myeloperoxidase, significantly inhibited both the binding and cytotoxicity of the serum. Finally, tumour necrosis factor-alpha, which is known to up-regulate cell surface expression of several ANCA antigens, enhanced both the binding and cytotoxicity of the serum. These findings suggest that ANCA induced by propylthiouracil contributed to leucopenia through a complement-mediated mechanism. PMID- 11882039 TI - The prevalence of autoantibodies in an elderly sub-Saharan African population. AB - In the present prospective, census-based study we have investigated the prevalence of organ-specific and non-organ-specific autoantibodies (AAb) in 152 unselected Cameroonians aged 60 years and older living in the community. AAb were detected in 49% of the participants. Non-organ-specific AAb (47%) predominated over organ-specific AAb (7%). Anti-TPO, anti-Tm, anti-Tg and anti-PC AAb were completely absent. RF was the most frequent AAb, being found in 57 (38%) cases. The prevalences of anti-SMA and RF were significantly higher in women than in men (respectively, P=0.023 and P=0.016). Higher serum concentrations of gammaglobulins were accompanied by a higher prevalence of RF (P < 0.0001) and a lower prevalence of ANA (P=0.036). The overall prevalence of AAb was higher in the filaria-infected (60%) compared to the non-infected (42%) participants (P=0.046). There was no significant influence of the vitamin D status, number of pregnancies, physical activity or medication use on the prevalence of AAb. In this study a heterogeneous pattern for the presence of the various AAb was found. Some AAb, which are commonly encountered in other studies on elderly subjects, were completely absent in this population. This diversified pattern of AAb prevalence therefore argues in favour of exogenous influences in the occurrence of AAb in elderly populations. PMID- 11882040 TI - Is ageing associated with a shift in the balance between Type 1 and Type 2 cytokines in humans? AB - The balance between Type 1 and Type 2 cytokines is important for the outcome of several infectious diseases. As elderly humans show increased morbidity and mortality from infectious diseases, this study tests if ageing is associated with a change towards Type 2 dominance in T cells. Expression of IFN-gamma, and IL-4 was measured in CD4+ and CD8+ T cells by flow cytometry in three groups: young controls (n=28), 81-year-olds (n=22), and centenarians (n=25). The major findings were that the percentage of IFN-gamma+ as well as IL-4+ T cells was increased in aged subjects. Furthermore, after adjusting for decreased lymphocyte counts in the elderly, the concentration in the blood of IFN-gamma+ and IL-4+ CD8+ T cells was still increased in the 81-year-olds. In centenarians, a shift towards a relative dominance of Type 2 cytokine expression was found within CD8+ T cells. Furthermore, the percentage of T cells with cytokine expression was closely correlated to the in vivo expression of CD95 and CD45RO. In conclusion, we found some evidence for an age-related shift towards a Type 2 cytokine profile. PMID- 11882041 TI - A hampered chemoattractant-induced cytoskeletal rearrangement in granulocytes of patients with unexplained severe chronic and relapsing infections of the upper and lower airways. In vitro restoration by G-CSF exposure. AB - Granulocytes play a major role in host defense against bacterial infections. Severe inborn defects in granulocyte function are associated with fulminant bacterial infections in early childhood. Subtle disturbances in granulocyte function might contribute to an enhanced susceptibility to bacterial infections in adulthood. We investigated chemoattractant (N-formyl-methionyl-leucyl phenylalanine, fMLP and casein) induced cytoskeletal rearrangements (polarization) of blood granulocytes in 77 adults with chronic and recurrent therapy-resistant infections of the upper and lower airways. These infections could not be explained by B- and/or T-cell defects or local anatomic abnormalities. Besides polarization, chemotaxis of blood granulocytes was measured in 33 patients, as well as granulocyte superoxide production in eight patients. The chemoattractant-induced cytoskeletal rearrangement in patient blood granulocytes was significantly lower as compared to healthy control values with both fMLP and casein as stimuli. About two-thirds of the patients showed a defective polarization response to fMLP. Granulocyte colony-stimulating factor (G CSF) when added in vitro corrected the defective polarization responses; responses in the normal range were not enhanced. The chemotactic motility of patient blood granulocytes was also slightly, but significantly lowered. However, it did not correlate to the lowered polarization. Granulocyte superoxide production was comparable in patients and in healthy controls. Our data thus show that subtle abnormalities in chemoattractant-induced cytoskeletal and motile function of blood granulocytes are frequent in patients with severe therapy refractory bacterial infections of the upper and lower airways. PMID- 11882042 TI - Increased soluble p55 and p75 tumour necrosis factor-alpha receptors in patients with hepatitis C-associated mixed cryoglobulinaemia. AB - To investigate whether tumour necrosis factor alpha (TNFalpha) plays a role in the pathogenesis of hepatitis C virus-associated mixed cryoglobulinaemia (HCV MC), we measured soluble TNFalpha and its soluble p55 (sTNFR1) and p75 (sTNFR2) receptors in the serum of patients with HCV-MC. TNFalpha, sTNFR1 and sTNFR2 were measured in the serum of 32 patients with HCV-MC, 18 patients with hepatitis C without MC (HCV) and 18 healthy volunteers, using specific immunoassays. Correlations between clinical and biological parameters and the concentrations of TNFalpha and sTNFRs were established by studying detailed clinical records of the 32 HCV-MC patients. Although higher, TNFalpha levels were not significantly different in HCV-MC patients compared with healthy or HCV controls. sTNFR1 and sTNFR2, however, were significantly higher in HCV-MC compared with controls or with HCV patients, and higher concentrations of sTNFR1 and sTNFR2 were observed in patients with severe visceral vasculitis, compared with patients with limited purpura. sTNFR1 concentrations positively correlated with fibrinogen levels but TNFalpha, sTNFR1 and sTNFR2 did not correlate with other biological parameters such as rheumatoid factor concentrations, CH50 or C4 values. These data suggest a role for TNFalpha in the pathogenesis of the immune complex-mediated vasculitis associated with HCV-MC. PMID- 11882043 TI - Complement C4 monitoring in the follow-up of chronic hepatitis C treatment. AB - The overall role of complement in the host--pathogen relationship is now well understood. However, its involvement at a chronic stage of infection, such as chronic hepatitis C, is less well documented. Here, results are reported which point to the use of specific C4 monitoring in the follow-up of HCV patients. This study concerns 66 patients with chronic HCV infection, treated with interferon alpha 2b alone or with interferon alpha 2b + ribavirin, and 50 healthy adults as controls. Complement blood tests were performed to measure C1q, C3, C4, mannan binding lectin (MBL), C1s-C1 inhibitor complexes, total (CH50) and C4 (C4H) haemolytic activity; specific C4 activity was taken as the C4H/C4 protein ratio. Rheumatoid factor (RF) levels were also measured. A significant reduction in CH50 and specific C4 activity in HCV patients, compared with the healthy controls, was observed before the onset of treatment; the other parameters were not affected and no C1s-C1 inhibitor complexes were detected. At the same time, a significant reduction in specific C4 activity was observed in relapsers compared with sustained responders. These results point to a potential predictive function of C4 specific activity to monitor the response to therapy. Restoration of specific C4 activity at 6 months was better in responders than in non-responders. Complement activation in chronic hepatitis C does not seem to involve the C1 stage of the classical pathway. A negative correlation between specific C4 activity and RF titres suggests a possible involvement of RF in C4 activation, via the lectin pathway. Specific C4 monitoring appears to be a valuable tool for the follow-up of chronic hepatitis C treatment, together with the other conventional investigations. PMID- 11882045 TI - Characterization of the phenotypic and lymphokine profile associated with strong CD8+ anti-HIV-1 suppressor activity (CASA). AB - A panel of 22 CD8+ T cell lines, with a broad range of CD8+ anti-HIV-1 suppressor activity (CASA) were generated from a single patient with HIV-1 infection. CD8+ T cell lines with either strong or weak CASA were examined and compared for cell surface and intracellular markers, constitutive chemokine and lymphokine mRNA levels and inducible lymphokine expression. Strong CASA significantly correlated with CD8+ T cell lines that highly coexpressed the molecule CD28+ (r=0.52, P=0.01) and Ki67+ (r=0.88, P=0.02), with strong CASA CD8+ T cell lines demonstrating significantly higher (P < 0.05) expression of CD8+CD28+ and CD8+Ki67+ compared to those with weak activity. No such correlations or findings were observed for the markers CD38, HLA-DR, CD57 or perforin. The Th1 cytokines were expressed at greater levels than the Th2 cytokines, with strong CASA significantly associated with an increased inducible level of IL-2 production (P=0.05). Constitutive RANTES, IP-10 and I-309 mRNA expression were significantly (P < 0.05) elevated in CD8+ T cell lines exhibiting strong CASA compared to those with weak CASA. There was no significant difference in the mRNA expression of the lymphokines IL-2, 4, 5, 8, 9, 10, 14, 15, or chemokines MIP-1alpha, MIP-1beta, MCP-1, and Ltn. Strong CASA was therefore associated with rapidly replicating CD8+ T cells of the phenotype CD8+CD28+Ki67+ that expressed greater levels of IL 2 and the ligands RANTES and I-309. PMID- 11882044 TI - The CD28/HLA-DR expressions on CD4+T but not CD8+T cells are significant predictors for progression to AIDS. AB - To investigate the changes of CD28 and HLA-DR molecules on CD4+ and CD8+ T cells during HIV infection, we classified 130 HIV-infected Koreans into four groups by the CD4 level as follows: group I (> or = 500 cells/mm3), group II (201-499 cells/mm3), group III (51-200 cells/mm3), and group IV (< or = 50 cells/mm3). In CD4+ T cells, the proportion of CD28 expression decreased significantly with the CD4 level while the proportion of HLA-DR expression increased gradually. In particular, the changes of HLA-DR expressions on CD4+ T cells were parallel to the loss of CD28 molecules from stage III to IV. However, the CD28 expression on CD8+ T cells decreased dramatically in the early stage of HIV infection, and the sum and pattern of CD28 and HLA-DR expressions on CD8+ T cells was stable after the first stage. Even though CD28 down-regulation on CD8+ T cells was very severe from the early stage of HIV infection, it might not influence the survival time of HIV-infected Koreans. The sum of the CD28+ subsets and HLA-DR subsets in each T cell was stable in all stages of disease progression. The sums of the CD28+ subsets and HLA-DR+ subsets in CD4+ T and CD8+ T cells were constant as approximately 100% and 55-60% of each T cell. These results suggested that the changes of CD28/HLA-DR expressions on CD4+ T cells were more predictable than those on CD8+ T cells in the evaluation of the disease progression during HIV infected periods. However, we need further studies to understand why the sum of two molecules in each T cell are constant. PMID- 11882046 TI - Cytokine production and apoptosis among T cells from patients under treatment for Plasmodium falciparum malaria. AB - Available evidence suggests that Plasmodium falciparum malaria causes activation and reallocation of T cells, and that these in vivo primed cells re-emerge into the periphery following drug therapy. Here we have examined the cytokine production capacity and susceptibility to programmed cell death of peripheral T cells during and after the period of antimalarial treatment. A high proportion of peripheral CD3+ cells had an activated phenotype at and shortly after time of admission (day 0) and initiation of therapy. This activation peaked around day 2, and at this time-point peripheral T cells from the patients could be induced to produce cytokines at conditions of limited cytokine response in cells from healthy control donors. Activated CD8hi and TCR-gammadelta+ cells were the primary IFN-gamma producers, whereas CD4+ cells constituted an important source of TNF-alpha. The proportion of apoptotic T cells was elevated at admission and peaked 2 days later, while susceptibility to activation-induced cell death in vitro remained increased for at least 1 week after admission. Taken together, the data are consistent with the concept of malaria-induced reallocation of activated T cells to sites of inflammation, followed by their release back into the peripheral blood where they undergo apoptotic death to re-establish immunological homeostasis as inflammation subsides. However, the high proportion of pre apoptotic cells from the time of admission suggests that apoptosis also contributes to the low frequency and number of T cells in the peripheral circulation during active disease. PMID- 11882047 TI - High prevalence of co-factor independent anticardiolipin antibodies in malaria exposed individuals. AB - Anticardiolipin antibodies (aCL) were investigated in 137 individuals chronically exposed to malaria and living in Africa and Asia. They belonged to several groups according to parasite (Plasmodium falciparum or vivax) and clinical manifestations (i.e. asymptomatic parasite carriers, acute uncomplicated attack or severe malaria episodes). aCL were measured in an enzyme immunoassay (ELISA) performed in the presence of either goat serum (aCLs) or gelatin (aCLg). In a group of 53 patients with autoimmune manifestations (i.e. antiphospholipid syndrome and/or lupus), detection of IgG but not IgM aCL was markedly reduced in the presence of gelatin. In malaria donors, high prevalence of serum co-factor independent IgG and IgM were detected, and the presence of goat serum in the assay consistently decreased their detection. aCLg levels were found to be related to the clinical/endemic status of donors. IgG aCLg were found to be higher in asymptomatic P. falciparum carriers than in patients with uncomplicated acute or cerebral malaria. IgM aCLg were higher in the cerebral malaria group than in groups with uncomplicated acute malaria patients or asymptomatic individuals. Data suggest that using a serum co-factor independent, sensitive ELISA, aCL are commonly detected during malarial infections and related to malarial infection status. PMID- 11882048 TI - Treatment with methylprednisolone in relapses of multiple sclerosis patients: immunological evidence of immediate and short-term but not long-lasting effects. AB - Relapses of multiple sclerosis (MS) are treated commonly with high-dose intravenous methylprednisolone (MP) given over a period of 3-5 days. The mechanisms responsible for the beneficial effects of MP in attacks are not clearly established. It is also controversial whether this treatment may have a long-term effect. Here, peripheral blood samples from relapsing--remitting MS patients in acute relapse were analysed by flow cytometry just before steroid treatment and at different time points after initiation of the therapy. We observed an immediate (day 3) decrease in the percentage of CD4+ lymphocytes, with a relative increase in the memory (CD4+CD45R0+) subpopulation. A longer standing effect of MP on IFN-gamma production, CD54, CCR5, CXCR3 and CD95 (Fas) expression was also observed on CD4+ cells after 1 month of treatment initiation. Six months after the therapy, during clinical remission, no changes due to ivMP therapy were detected. These results support that MP treatment of relapses induces immediate post-treatment and short-term effects on the immune system that could partly account for the clinical and radiological improvement observed in MS patients. However, no conclusion can be drawn as to a possible long-term or even intermediate influence of ivMP treatment on the course of the disease. PMID- 11882049 TI - Anti-multiple nuclear dots (anti-MND) and anti-SP100 antibodies in hepatic and rheumatological disorders. AB - Multiple nuclear dots pattern has been described in primary biliary cirrhosis and, less often, in rheumatological disorders. Sp100 is the major antigen of multiple nuclear dots. We evaluated prevalence and diagnostic significance of multiple nuclear dots and anti-Sp100 reactivity both in hepatic and rheumatological diseases. A series of 283 consecutive liver patients (89 primary biliary cirrhosis, 12 primary sclerosing cholangitis, 85 autoimmune hepatitis, 97 hepatitis C virus-related chronic liver disease) and of 89 consecutive rheumatological cases were evaluated. Presence of multiple nuclear dots was assessed by indirect immunofluorescence on HEp-2 cells, anti-Sp100 reactivity by ELISA with recombinant protein. Multiple nuclear dots were detected in 20 patients (7%) with liver disease (of whom 15 with primary biliary cirrhosis), and in eight patients (9%) with rheumatological disorders. Anti-Sp100 was detected in 45 liver patients (16%), of whom 30 with primary biliary cirrhosis, but in only two with rheumatological disorders (2%) (P =0.0004). The concordance between multiple nuclear dots and anti-Sp100 in liver and rheumatological patients was 90% and 25% (P=0.0018), respectively. Among 89 consecutive patients with primary biliary cirrhosis, multiple nuclear dots and anti-Sp100 were present in 17% and 34%, respectively (P=0.0152). Anti-Sp100 positivity was associated with older age and higher gamma-globulin levels. Multiple nuclear dots are similarly observed in liver and rheumatological patients. In contrast, anti-Sp100 is more frequent in liver patients and is significantly more often detected in primary biliary cirrhosis, of which it can be regarded as a highly specific serological marker. The antigenic target of multiple nuclear dots in most rheumatological patients is other than Sp100. PMID- 11882051 TI - Death-inducing receptors and apoptotic changes in lymphocytes of patients with heart transplant vasculopathy. AB - The specific role of lymphocyte apoptosis and transplant-associated atherosclerosis is not well understood. The aim of our study was to investigate the impact of T cell apoptotic pathways in patients with heart transplant vasculopathy. Amongst 40 patients with cardiac heart failure class IV who have undergone heart transplantation, 20 recipients with transplant-associated coronary artery disease (TACAD) and 20 with non-TACAD were investigated one year postoperative. Expression of CD95 and CD45RO, and annexin V binding were measured by FACS. Soluble CD95, sCD95 ligand (sCD95L), tumour necrosis factor receptor type 1 (sTNFR1), and histones were measured in the sera by ELISA. The percentage of cells expressing CD3 and CD4 was significantly reduced in TACAD as well as in non-TACAD patients as compared with control volunteers. Interestingly, the proportion of CD19+ (B cells) and CD56+ (NK) cells was increased in TACAD groups (versus non-TACAD; P < 0.01, and P < 0.001, respectively). In contrast to sCD95, the expression of CD95 (APO-1/Fas) and CD45RO (memory T cells), and sCD95L were significantly increased in non-TACAD and TACAD patients. T cell activation via CD95 with consecutive apoptosis was increased in both groups. The concentration of sTNFR1, IL-10 and histones was significantly elevated in sera from TACAD than non-TACAD patients, and in both groups than in healthy controls. These observations indicate that the allograft may induce a pronounced susceptibility of CD4+ T cells to undergo apoptosis and antibody-driven activation-induced cell death. This data may suggest a paradox immune response similar to that seen in patients with autoimmune diseases. PMID- 11882052 TI - Reporting clinical trials: full access to all the data. PMID- 11882053 TI - Clinical features and assessment of severe dementia. A review. AB - Sound understanding of the dementia syndrome requires adequate acquaintance with its entire spectrum, from the lightest to the most advanced stages. Most studies of dementia deal with light to moderate stages of the condition, while relatively little attention has been paid to its most severe stages. This review presents a clinical description of patients with severe dementia and of the tests currently available to evaluate their cognitive, behavioural, and functional status. Available instruments such as the Hierarchic Dementia Scale or the Severe Impairment Battery now allow quantification of the cognitive and behavioural status of patients with severe dementia. Experience with severe dementia shows that, far from being in a "vegetative state", as is commonly thought, late-stage patients are in fact quite different from one another and in most cases continue to have an interaction with their environment. This ability to better define the characteristics of patients with severe dementia provides the basis for correlations between clinical data and data derived from neuroimaging, neurochemistry, or neuropathology. It also sets the stage for possible therapeutic trials involving these patients. PMID- 11882050 TI - CD3+CD56+ NK T cells are significantly decreased in the peripheral blood of patients with psoriasis. AB - Psoriasis is a chronic, inflammatory, hyperproliferative skin disease, in which autoimmunity plays a great role. Natural killer T cells (NK T cells), are suggested to be involved in the pathogenesis of different autoimmune diseases. To examine the involvement of CD3+CD56+ NK T cells in the pathogenesis of psoriasis, we investigated the lymphocyte subpopulations obtained from blood samples of psoriatic patients before and after treatment, and of healthy controls, using two colour flow cytometry. We found no significant differences between total T cells, total B cells, T helper cells, T cytotoxic cells and NK cells in patients with psoriasis before and after treatment and in controls. Increased percentage of memory T cells and decreased percentage of naive T cells was detected in psoriatic patients compared to controls, but these changes were not statistically significant. The CD3+CD56+ cells of psoriatic patients were significantly decreased relative to controls. The percentage of CD3+CD56+ cells increased after different antipsoriatic therapies, but remained significantly lower than those found in controls. CD3+CD56+ cells of healthy controls were capable of rapid activation, while in psoriatic patients activated NK T cells were almost absent. The decrease in the number of CD3+CD56+ cells may represent an intrinsic characteristic feature of patients with psoriasis, which is supported by the fact that after treatment NK T cells do not reach the values found in controls. In conclusion our results suggest that CD3+CD56+ NK T cells could be actively involved in the development of Th1 mediated autoimmune diseases. PMID- 11882054 TI - Impaired benzodiazepine receptor binding in peri-lesional cortex of patients with symptomatic epilepsies studied by [(11)C]-flumazenil PET. AB - Individual benzodiazepine receptor (BZR) binding of peri-lesional cortex was investigated in symptomatic epilepsies. Eleven patients aged 19-44 years were studied whose diagnosis was established by medical history, clinical, electroencephalographic, and magnetic resonance imaging (MRI) findings. Three dimensional [11C]-flumazenil (FMZ) positron emission tomography and MRI scans were obtained and coregistered. Lesions (five low-grade brain tumours, one AV malformation, one cavernoma, one cystic lesion of unknown aetiology, one traumatic brain injury, one post-operative and one post-haemorrhagic defect) were outlined on individual MRI scans. Adjacent to those lesions, and in homologous contralateral structures, FMZ binding was analysed in four pairs of cortical 9 x 9-mm regions of interest (ROIs) placed on transaxial and coronal slices, respectively, as well as in the lesion volume and its mirror region. Percentage asymmetry ratios were calculated and those at or outside the 90-110% range were operationally defined significant. Peri-lesional FMZ binding asymmetries ranged from 70 to 125%, lesional asymmetries from 38 to 82%. Only one patient showed no significant change, whilst nine exhibited significant reductions of FMZ binding in at least one ROI (3 x 1, 4 x 2, 1 x 3, 1 x 4), and significant increases were observed in two ROIs of another patient. Therefore, peri-lesional disturbances of BZR binding are common but variable in location. Because a close correlation between regional decreases in FMZ binding and spiking activity was recently demonstrated in neocortical epilepsies, abnormal peri-lesional FMZ binding may bear some relation to the mechanisms of epileptogenesis in symptomatic epilepsies. PMID- 11882055 TI - Neuropsychological deficits in essential tremor: an expression of cerebello thalamo-cortical pathophysiology? AB - Few studies have been published regarding the neuropsychological characteristics of patients with essential tremor (ET), but preliminary findings suggest that mild attentional and executive dysfunction accompany the disorder. A consecutive series of 101 patients with ET referred for thalamotomy and/or thalamic deep brain stimulation candidacy work-up also underwent neuropsychological evaluation. Average neuropsychological test scores were calculated, along with the proportions of subjects whose scores fell within or more than one SD above or below the mean (using demographically corrected normative data). Significantly lower than average (T-score of 50) scores were evident on measures of complex auditory attention, visual attention and response inhibition, recall of a word list, verbal fluency, and visual confrontation naming. A significantly greater proportion of patients (ranging from about 34 to 60%) than might be expected on the basis of a normal distribution obtained scores more than one SD below the normative mean on select measures of attention, verbal fluency, immediate word list recall, semantic encoding, and facial matching. Consistent with prior research, notable, albeit clinically subtle, deficits in attention and select executive functions are evident in patients with ET. Although not specific to ET or cerebellar dysfunction, the observed pattern of cognitive deficits is consistent with cerebello-thalamo-cortical circuit dysfunction. PMID- 11882057 TI - Therapeutical considerations in patients with intracranial venous angiomas. AB - The clinical presentation of intracranial venous angiomas are usually headache, seizures or dizziness. Very often these anomalies are found incidentally on magnetic resonance imaging (MRI). We reviewed 35 patients, which were examined in our department between 1994 and 2000. Only one patient became symptomatic with an intracranial haematoma, which was removed surgically. In three patients an associated cavernous angioma was found, which was removed successfully with preservation of the coexisting venous angioma. There is no indication in operating a venous angioma because the risk of postoperative deterioration caused by venous infarction is high. When removing cavernous angiomas the associated venous angioma has to be preserved. PMID- 11882056 TI - Depletion of Vbeta5.2/5.3 T cells with a humanized antibody in patients with multiple sclerosis. AB - A potentially pathogenic expansion of T cells expressing T cell receptor (TCR) Vbeta5.2/5.3 has been demonstrated in patients with multiple sclerosis (MS). A humanized antibody (ATM-027) directed against these T cells has been developed to further investigate the role of this subpopulation of T cells in MS. The pharmacokinetics/dynamics and safety of ATM-027 (0.3-300 mg intravenously over 30 min) were investigated in 14 patients with MS. The effect of treatment on cytokine expression and autoreactivity to peptides of myelin basic protein (MBP) was also studied. ATM-027 was well tolerated and raised no safety concerns. Clearance of the antibody was low and elimination half-life was approximately 3 weeks. The majority of the target Vbeta5.2/5.3 expressing T cells were depleted for at least 18 months. The small remaining fraction of target cells showed a marked decrease in their TCR expression, which was recovered within 8 months. The numbers of peripheral blood mononuclear cells (PBMCs) with spontaneous expression of IFN-gamma was decreased at 72 h and 8 weeks after treatment, whilst no clear effects on TNF-alpha, IL-4, IL-10, TGF-beta expression were observed. There was also a significant decrease in the number of PBMCs producing IFN-gamma in response to MBP peptide 80-102. We conclude that long-term depletion of T cells expressing defined Vbeta subgroups in MS patients is feasible using selective immunotherapy. The selective depletion of Vbeta5.2/5.3 expressing T cells in this study resulted in a decrease in potentially disease promoting anti-MBP reactivity and pro-inflammatory cytokine production. PMID- 11882058 TI - Epidemiological aspects of headache in a workplace setting and the impact on the economic loss. AB - The objective of this study was to assess the epidemiological aspects of headache in a workplace setting and the economical impact. By sending a questionnaire to 1781 employees of a Dutch manufacturing company we tried to assess the prevalence of headache and headache subtype within this working place. Besides the personal view of each responder regarding his/her headache subtype, the International Headache Society (IHS) criteria were used to classify headaches on the basis of reported features. Moreover the missing labour days, as a result of headache, during the previous 4 weeks were measured, as well as the loss of productivity at the working place during an episode of headache; from this an estimate of the economic loss could be calculated. The response rate of the questionnaire was 60.8%. The lifetime prevalence of headache amongst the responders was 53.3%. Of these, 34.5% thought their headache was tension-type headache, 10.6% thought it was migraine, 5.4% mentioned daily headache, and 18.2% mentioned another type of headache. According to the IHS criteria the differentiation was somewhat different: 26.2% of the headache could be classified as tension-type headache and 15% as migraine. When the costs for lost labour days and loss of productivity as a result of all types of headache were accumulated, the economic loss was estimated at 18.933 US dollars in 4 weeks (for the group of responders). In conclusion, headache is related to a substantial economic loss; migraine is probably underestimated in a workplace setting. PMID- 11882059 TI - Complex hallucinations following occipital lobe damage. AB - Visual hallucinations may accompany many neurological and psychiatric disorders. A common localization principle is that lesions to the early sensory cortices lead to elementary hallucinations, whereas complex perceptual experiences are related to the pathology of higher-level cortical regions. We report the case of a patient who experienced complex, non-stereotyped, multimodal (visual and somatosensory) hallucinations following an acute ischaemic vascular lesion in the right medial occipital lobe. This illustrates that the phenomenology of hallucinations not necessarily reflects the exact localization of cerebral pathology. Instead, the damaged area may serve as a focus of an abnormally activated neuronal network. PMID- 11882060 TI - Relapsing-remitting demyelinating illness: multiple sclerosis, multiphasic disseminated encephalomyelitis, or an intermediate entity? PMID- 11882061 TI - Non-convulsive status epilepticus of frontal origin following surgery for craniopharyngioma. PMID- 11882062 TI - HLA-DR15 and age at onset in multiple sclerosis. PMID- 11882063 TI - A case of concomitant amyotrophic lateral sclerosis and HIV infection. PMID- 11882064 TI - Risperidone in chorea and psychosis of Huntington's disease. PMID- 11882066 TI - Zolpidem in progressive supranuclear palsy. PMID- 11882065 TI - Riluzole and olanzapine in Huntington's disease. PMID- 11882067 TI - Amyloid neuropathy presenting as thoraco-abdominal parathesia. PMID- 11882074 TI - Management of spontaneous inhibitors in children with porcine factor VIII. AB - Acquired factor VIII (FVIII) inhibitors are rare, typically occurring in the postpartum period or in the elderly. Their occurrence in childhood is distinctly unusual. Acquired FVIII inhibitors are often life-threatening and refractory to treatment with high doses of human FVIII concentrate. Alternative strategies for control of haemostasis include the use of products with FVIII "bypassing" activity or porcine FVIII (pFVIII) concentrate if the pFVIII titre is sufficiently low (<10-20 porcine Bethesda Units). Corticosteroids and other immunosuppressive therapies are inconsistently effective in eliminating FVIII inhibitors. Accordingly, acquired FVIII inhibitors often require long-term haemostatic management. PMID- 11882075 TI - Porcine factor VIII in the treatment of high-titre inhibitor patients. AB - Development of an inhibitor against factor VIII (FVIII) is an important complication of haemophilia. It occurs in approximately 25-30% of patients with haemophilia A [1]. FVIII inhibitors may also occur as autoantibodies. The latter occur in non-haemophiliacs and, although rare (occurring in approximately one per million of the population), are frequently associated with life-threatening bleeding. Inhibitors are considered low level if they are < 5 Bethesda Units (BU) or high level if they are > 10 BU. The former usually remain low and rarely give anamnestic response, the latter do so frequently. Despite various approaches to their management, the presence of inhibitors remains a major cause of morbidity and mortality. The effectiveness of porcine FVIII (pFVIII) in treating patients with both auto- and alloantibodies to FVIII has been well demonstrated when adequate circulating levels of FVIII are obtained [2--10]. However, pFVIII therapy may give rise to antibodies to the pFVIII and the utility of this treatment in the presence of high levels of porcine antibody is less well recognized and understood. Nonetheless, pFVIII under these circumstances may be useful in a select group of patients where management is difficult. PMID- 11882076 TI - Continuous infusion of porcine factor VIII: stability, microbiological safety and clinical experience. AB - Porcine factor VIII (pFVIII) is an effective haemostatic treatment for bleeding in selected patients with FVIII inhibitors. Its use is sometimes associated with a transient fall in platelet count and transfusion reactions, the risk of which may be related to the rate of administration. Theoretical considerations suggest that the administration of pFVIII by continuous infusion should be effective, and could have pharmacokinetic advantages that lead to an improvement in the side effect profile. The results of a retrospective survey of continuous infusion of pFVIII with respect to clinical safety and efficacy are reported. Porcine FVIII stability and microbiological studies are included. It is concluded that pFVIII given by continuous infusion is safe and effective. The risk of transfusion reactions and fall in platelet count appears to be reduced, compared with bolus administration. Stability studies showed that pFVIII activity declined at room temperature, most rapidly in the dilute solution (5-10 U mL(-1)). More concentrated mixtures showed acceptable stability for up to 24 h using a variety of infusion devices. Various concentrations of pFVIII did not support the growth of Escherichia coli or Staphylococcus aureus. These observations suggest that the porcine factor is suitable for continuous infusion (CI). PMID- 11882077 TI - Porcine factor VIII: pharmacoeconomics of inhibitor therapy. AB - The treatment of acquired haemophilia is characteristically exceedingly expensive and thus a cost-benefit analysis of the several available treatment strategies is urgently needed. To address this issue, decision-analysis techniques were used to construct a cost-minimization model to compare the cost of treatment with porcine factor VIII (pFVIII), human FVIII (hFVIII) or an activated prothrombin complex concentrate (APCC). This model was based upon the results of a comprehensive literature search of all relevant clinical studies and case series. To supplement these data, a panel of haemophilia specialists was presented with a clinical scenario describing an acquired haemophilia patient with an acute haemorrhage in whom the human and porcine inhibitor titres were initially unknown. Based on this scenario and on their own clinical experience, the expert panel assessed the applicability of the model as initially constructed, assigned probabilities of success to each treatment and recommended appropriate initial dosing and follow up regimens. This information was incorporated into the model and a simulation was conducted from which the costs of care were calculated. Sensitivity analyses were then conducted on all parameters. The results of the model show that treatment initiated with pFVIII would be more cost effective compared with treatment sequences initiated with an APCC or hFVIII, respectively. The model indicates that initial treatment with pFVIII in this scenario may be the preferred strategy clinically, as well as on economic grounds. PMID- 11882078 TI - The role of porcine factor VIII in the management of unexpected bleeding episodes. AB - The introduction of porcine factor VIII (pFVIII) has been an important addition to our armamentarium in the management of patients with inhibitors to FVIII. Case studies will be presented that offer important differential diagnoses. The prevalence and indications for the use of this biological replacement therapy will be discussed. PMID- 11882079 TI - Use of Protein-A column and porcine factor VIII. AB - Extracorporeal immunoadsorption of factor VIII (FVIII) antibodies using Sepharose matrix columns coupled with staphylococcal Protein-A was reported two decades ago. The efficiency of this technique for removing FVIII antibodies of the IgG subtypes was clearly demonstrated. The recent widespread use of a variety of apheresis techniques for the management of a multitude of haematological and oncological conditions has made this technology more accessible and affordable. For the treatment of patients with FVIII inhibitors, the use of porcine FVIII makes it possible to control haemostasis with a therapeutic product for which in vitro testing can help predict the in vivo efficacy. By lowering the level of FVIII inhibitors, immunoadsorption can make the use of pFVIII concentrate possible in situations otherwise untreatable with FVIII preparations. Moreover, lowering the level of FVIII inhibitors by immunoadsorption allows adequate haemostasis to be achieved with much lower doses of FVIII leading to significant saving. Our preliminary data suggest that immunoadsorption combined with the use of pFVIII should be considered early in the treatment plan for controlling haemostasis in patients with FVIII inhibitors. PMID- 11882080 TI - Porcine factor VIII: current status and future developments. AB - High-purity porcine factor VIII (pFVIII) (Hyate:C, Ipsen, UK), with a specific activity of > 125 U mg(-1) protein, has been shown to be effective in up to 90% of bleeds in patients with FVIII inhibitors. These inhibitors have been shown to have a median 15% cross-reactivity to pFVIII, and even less cross-reactivity in patients with acquired haemophilia. Its use is sometimes associated with a transient fall in platelet count and with transfusion reactions. Furthermore, it is not virally attenuated. Although pFVIII has not been shown to transmit any viral illness to its human recipients, it is extensively screened for porcine viruses using a 4-cell line general screen. The source plasma is also screened for porcine parvovirus. To satisfy the demand for ever-improved side-effect profile and viral safety a third-generation pFVIII is under consideration by Ipsen. This product will be purified from porcine plasma screened for porcine parvovirus, using immuno-purification, ion-exchange chromatography and washing. Spiking experiments using human FVIIIC (hFVIIIC) suggest that these purification steps may be associated with a 6-log viral reduction. The product is also virally attenuated using the solvent-detergent method. This should yield an ultra-pure concentrate, lacking porcine von Willebrand factor, with a specific activity of approximately 5000 U mg(-1) protein which does not require added albumin as stabilizer and which may be stored at 2-4 degrees. One would anticipate that this product should be clinically effective and should be associated with enhanced viral safety and an improved side-effect profile. It should not affect the platelet count and should be associated with a much reduced risk of transfusion reactions. Clinical trials are planned. PMID- 11882082 TI - Single-dose pharmacokinetics of porcine factor VIII (Hyate C). AB - The objective was to establish the pharmacokinetic properties of porcine factor VIII (pFVIII) at three different doses in a single patient. The patient, born in 1952, had severe haemophilia A and developed an inhibitor to human FVIII (hFVIII) in 1966 aged 14 years. He was first treated with pFVIII in 1980. Apart from a short period of treatment with hFVIII in 1998 which resulted in the reappearance of the inhibitor, pFVIII has been constantly used since 1984. No inhibitor against human or porcine FVIII had been recordable over the 2-year period prior to the study. Three separate pharmacokinetic studies were performed using a washout period of 72 h and doses of 10 U kg (-1), 25 U kg (-1), or 50 U kg (-1), respectively, with sampling at preinfusion, then at 15 and 30 min and 1, 3, 6, 9, 12 and 24 h postinfusion. The FVIII levels were measured using both plasma derived one stage APTT based assay and chromogenic assay. The results were computed using a model-independent analysis and a model-dependent analysis. The respective clearances (mL h(-1)kg(-1)) at doses 50 U, 25 U or 10 U kg (-1) were 1.32, 1.33 1.8 (bioassay) and 2.54, 2.93, 9.43 (chromogenic). The respective half lives (h) at doses 50 U, 25 U, and 10 U kg (-1) were 23.71, 16.54, 25.17 (bioassay) and 15.71, 17.39, and 10.66 (chromogenic). The respective recoveries (u dL(-1)/u kg(-1)) at doses 50 U, 25 U, and 10 U kg (-1) were 2.32, 2.44, 2.7 (bioassay) and 1.42, 1.16 and 0.9 (chromogenic). It was found that the two compartment model best fitted the curves of the bioassay and a one compartmental model best fitted the curves of the chromogenic assay. The pharmacokinetic studies are the first to be performed at different dose levels and using different assay methods for pFVIII. Using the bioassay, they show a long half life and high recovery compared to hFVIII. The differences between the bioassay and the chromogenic assay reflect their different biological basis and are of relevance when potency labelling is performed using chromogenic assay (European Pharmacopeia). PMID- 11882084 TI - Guest editorial: 10 years of the 'new' community care: good in parts? PMID- 11882085 TI - The school health nurse's assessment of a successful health dialogue. AB - The purpose of this paper is to achieve a deeper understanding of the learning processes that occur within a regular health dialogue between pupils and school health nurse from the school health nurse's perspective. The study used a grounded theory approach for the data collection and analysis. Eight experienced school health nurses were interviewed using a semistructured 'interview guide'. The interviews were audio recorded and transcribed verbatim. A detailed analysis presented the processes involved and the core category emerged as creating a communicative room to learn about health. The core category contained factors related to competencies, authenticity, reflective openness and environmental support, which was the basis for learning processes and a successful health dialogue. This paper adds to our understanding of the processes in school health nursing and the learning about health which takes place within a health dialogue. PMID- 11882086 TI - Are primary care groups and trusts consulting local communities? AB - Primary care groups and trusts (PCG/Ts) in the English NHS were established in 1999 and have responsibility for providing and commissioning health-care for around 100 000 people. PCG/Ts are dominated by health professionals, but are responsible for representing the interests of the local community. This paper assesses how they have informed and consulted local communities and the perceived impact of this consultation on decision-making. The paper uses evidence from the National Tracker Survey of PCG/Ts, a longitudinal survey of 72 (15%) of the PCG/Ts in England, using data from telephone interviews with chairs and chief officers, and postal questionnaires to lay board members and representatives of Community Health Councils (CHCs). Eighty-one per cent of PCG/Ts had public involvement working groups. Methods of consulting the community included consulting CHCs (87%), holding public meetings (75%) and consulting local patient groups (67%). Only 31% of chairs felt they were effective at consulting. Ninety two per cent of CHC representatives attended all board meetings. Most CHC representatives reported that there had been little or no consultation with the CHC in areas such as commissioning, service development or clinical governance. Only 14% of CHC representatives rated PCG/T consultation with the public as effective. Eighty-seven per cent said that local communities were largely unaware of the existence of PCG/Ts, and 70% commented on the weaknesses in PCG/T efforts at public consultation. Public participation is being taken seriously by PCG/Ts, but most are struggling to develop effective ways of involving local communities. Efforts to involve the public may become little more than token gestures. The proposed abolition of CHCs may make it more difficult for PCG/Ts to obtain a lay perspective. Effective consultation requires the development of new methods and adequate resources, but a stronger lay voice in the governance structures of PCG/Ts is needed. PMID- 11882087 TI - Using network variation in practice: identification of support network type. AB - This paper evaluates the use of a support network measurement instrument (PANT) in social work practice, focusing primarily on a study of the introduction and use of the instrument with social work teams. The paper describes practitioners' responses during the training period and during the initial phases of using the instrument in practice. The findings show how growth in familiarity with the new tool and support from managers overcame early resistance and contributed to both practitioners' confidence and understanding of the situations of older people. It allowed them to tailor interventions appropriately. It also contributed to understanding of differences between teams in the area and made it possible to create area profiles of older service users, their carers and their needs. The importance of network type for prediction of outcomes is stressed. PMID- 11882088 TI - Going home from hospital -- an appreciative inquiry study. AB - This paper reports on a project that involved a number of agencies and groups, including older people, working together to examine and develop practice in an area of shared concern -- going home from hospital. The project was stimulated by a 'whole-system event', and was based on appreciative inquiry (AI) methodology, which has roots in both action research and organisational development. In AI, the research is directed towards appreciating what it is about the social world that is positive, and exploring this. The study was planned around three workshops to streamline data collection and analysis. Group members were also required to carry out some activities between workshops. Invitations were sent out to groups and individuals previously identified as involved or interested in the discharge process across one health district (n = 71). Workshop one discussed the planned research schedule, and introduced the basic concepts of AI. This workshop also took participants through the interview process. Each participant was asked to undertake two interviews. Thirty-five individual interviews and one focus group were completed. At workshop two, interview data were analysed by the group using the nominal group technique. Subsequent group discussion produced 'provocative propositions'. At the third workshop, provocative propositions were developed into action plans. This paper gives an overview of the study, and explores some of the issues involved when working with service users and providers as co-researchers. PMID- 11882089 TI - The early days of primary care groups: general practitioners' perceptions. AB - English primary care is currently undergoing radical reform. Primary care groups (PCGs), effectively compulsory federations of general practices, came into legal existence in April 1999. This paper contains a review of general practitioners' (GPs') initial perceptions of the impact of these reforms on practice and considers the wider issue of changes in professional autonomy. A random sample of 49 GPs from two adjacent health districts in the North of England were interviewed as part of a longitudinal qualitative study. One round of interviews took place 7 to 9 months after the creation of PCGs and a further round of interviews was carried out 6 months later. We were interested in GPs' knowledge of recently formed (PCGs') plans and priorities, the impact of PCG activity to date, and the predicted future impact of such activity. After the published priorities of PCGs had been identified, thematic content analysis was used to ascertain GPs' perceptions of those priorities. GPs were generally unaware of their PCGs' published priorities. The wider strategic role of PCGs in commissioning services was rarely alluded to. Although over a third of GPs reported no current impact of the PCG, the majority expected PCGs to have considerable impact. In particular, control, management and accountability arrangements were all perceived as central issues in the expected developments. Performance management arrangements related to specific clinical priorities were widely expected. Although the new arrangements were inspiring little enthusiasm, the reforms did not appear to threaten GPs sufficiently enough to provoke active resistance. PMID- 11882096 TI - Reorganizing the delivery of intensive care could improve efficiency and save lives. AB - Critically ill patients are at high risk for death and permanent disability. Their care is also very expensive. The method of delivering critical care services to these patients can have an impact on their clinical and economic outcomes. Researchers face significant challenges when attempting to identify an association between an intensive care unit (ICU) organizational characteristic and patient outcomes. In this paper we review the challenges faced when evaluating the impact of ICU organizational characteristics on patient outcomes and highlight ICU characteristics that are consistently associated with improved patient outcomes. These characteristics include: (i) the presence of specialist physicians devoted to the ICU; (ii) increased nurse : patient ratios; (iii) decreased use of tests and evaluations that will not change clinical management; (iv) development and implementation of evidence-based protocols and guidelines; (v) use of computer-based alerting and reminding systems; and (vi) having a pharmacist participate in daily rounds in the ICU. When implementing these in ICUs, it is important to evaluate the impact of these characteristics on patient outcomes. We provide a format for such an evaluation. Given the growing evidence supporting the association between specific ICU characteristics and improved patient outcomes, we hope the future realizes broad implementation of these beneficial characteristics. PMID- 11882097 TI - Quantitative evaluation of prostatectomy for benign prostatic hypertrophy under a national health insurance law: a multi-centre study. AB - Assessing regional variation between various medical centres in diagnostic and surgical processes is an approach aimed at evaluating the quality of care. This study analyses the differences between eight medical centres in Israel, where all citizens are covered by medical insurance, through the National Health Insurance Law (NHIL). The analysis refers to the diagnostic process, type of surgery and immediate post-surgical complications associated with prostatectomy for benign prostatic hypertrophy (BPH), which is the most frequent surgical procedure performed on men aged 50+. The study sample was comprised of 261 consecutive prostatectomy patients operated on in eight Israeli medical centres (MC), located in various parts of the country, between November 1996 and April 1997. Co operation with participating directors of surgical wards was obtained after confidentiality of information had been assured. Surgeons in selected departments abstracted data routinely recorded in the patient's file and filled-out a standard one-page questionnaire. The following items were included: age, the presence of accompanying chronic diseases, preoperative tests, type of operation, and post-surgical complications. In the various MCs 32.6% of the patients underwent more than five preoperative tests ranging from 8.9% to 88.9% (<0.01). Assessment of kidney and bladder normality ranged from 75% to 100% (P < 0.01). The rate of patients whose prostatic symptoms (I-PSS) were assessed ranged from 0% to 79% (P < 0.01). There were also differences in severity of prostatism between the MCs, with severe symptoms ranging from 54.0% to 89.3% (P < 0.05), for type of operation performed (for 'open' prostatectomies, 35.4% to 68.0%, P < 0.01) and post-operative complications (19.0% to 41.6%, P = 0.07). After controlling for case-mix, type of operation was the most important predictor for post-surgical complications. MCs with low volume of surgeries had a higher rate of postoperative complications. We conclude that diagnostic and type of operation and post-surgical complications differed between various MCs. Participating surgeons were willing to fill out a one-page standard questionnaire from data routinely recorded in patients' files. PMID- 11882098 TI - Family physicians' and general practitioners' approaches to drug management of diabetic hypertension in primary care. AB - RATIONALE, AIMS AND OBJECTIVES: To compare the pharmacotherapeutic approaches to diabetic hypertension of family physicians (FPs) and general practitioners (GPs). METHODS: A retrospective prescription-based study was conducted in 15 out of a total of 20 health centres, involving 115 primary care physicians--77 FPs and 38 GPs, representing 74% of the primary care physicians of Bahrain. Prescriptions were collected during May and June 2000 to comprise a study population of 1266 diabetic-hypertensive patients. RESULTS: As monotherapy, angiotensin-converting enzyme (ACE) inhibitors (37.9%) and beta-blockers (38.3%) were the most commonly prescribed classes of antihypertensives by FPs and GPs, respectively. Calcium channel blockers (CCBs) were ranked third by both categories of physicians. For two-drug combinations, a beta-blocker and an ACE inhibitor was the combination of choice for both physician categories. Patients managed by the FPs were more likely to receive a beta-blocker-CCB combination (17.4 vs. 14.9%) or a diuretic ACE inhibitor combination (16.7 vs. 11.4%) and less likely to receive a beta blocker-diuretic combination (11.8 vs. 16.7%) than those managed by the GPs. The proportion of patients receiving antihypertensive combinations was 40.6 and 38.5% for FPs and GPs, respectively. While the GPs prescribed CCB as a monotherapy to the elderly most often, the FPs choice was a beta-blocker. Diuretics were less preferred by both FPs and GPs. Beta-blocker-ACE inhibitor was again the most preferred combination of both FPs and GPs. FPs prescribed CCB-beta-blocker combinations more often than GPs (P = 0.01), whereas CCB-ACE inhibitor combinations were less preferred (P = 0.09). A trend towards excessive use of short-acting nifedipine as monotherapy for elderly patients, both by FPs and by GPs, was noticed. Glibenclamide, alone or in combination with metformin, was the foremost antidiabetic drug prescribed by FPs and GPs. Middle-aged (45-64 years) patients seen by GPs were more likely to receive glibenclamide than those treated by FPs (P = 0.001) and less likely to receive gliclazide (P = 0.01). Combinations of a beta-blocker with either glibenclamide or insulin were prescribed considerably more often by GPs. CONCLUSIONS: Within the same practice setting, a substantial difference was observed between FPs and GPs in terms of preference for different classes of drugs in the management of diabetic hypertension. The compliance of both FPs and GPs was suboptimal; overall, the compliance of the FPs was closer to the recommended guidelines, however. Educational programmes should specifically address these inadequacies in order to improve the quality of health care. PMID- 11882099 TI - Telephone reporting in the consultant-generalist relationship. AB - RATIONALE, AIMS AND OBJECTIVES: The aim of this paper is to examine the issue of telephone reporting by consultants in the medical community, which is rarely used in medicine today. METHODS: The paper reviews principal publications addressing this field that have shown that communication between consultants and primary care physicians has many shortcomings, frequently leaving the primary care physician with unanswered questions. DISCUSSION AND CONCLUSIONS: We present the value of routine telephone reporting by consultants to their primary care counterparts and also discuss possible future implications of communication via electronic mail. PMID- 11882100 TI - The effect of participation in a weight loss programme on short-term health resource utilization. AB - Obese people consume significantly greater amounts of health resources. This study set out to determine if health resource utilization by obese people decreases after losing weight in a comprehensive medically supervised weight management programme. Four hundred and fifty-six patients enrolled in a single centred, multifaceted weight loss programme in a universal health care system were studied. Patient information was anonymously linked with administrative databases to measure health resource utilization for 1 year before and after the programme. Mean body mass index (BMI) decreased by more than 15%. The mean annual physician visits (pre = 9.6, post = 9.4) did not change significantly after the programme. However, patients saw a significantly fewer number of different physicians per year following the programme (pre = 4.5, post = 3.9; P < 0.001). Mean annual number of emergency visits (pre = 0.2; post = 0.2) and hospital admissions (pre = 0.05; post = 0.08) did not change. Neither baseline BMI, nor its change during the programme, influenced changes in health resource utilization. Our study suggests that weight loss in a supervised weight management programme does not necessarily decrease short-term health resource utilization. Further study is required to determine if patients who maintain their weight loss experience a decrease in health utilization. PMID- 11882101 TI - The 2-year costs and effects of a public health nursing case management intervention on mood-disordered single parents on social assistance. AB - RATIONALE, AIMS AND OBJECTIVES: This randomized controlled trial was designed to evaluate the 2-year costs and effects of a proactive, public health nursing case management approach compared with a self-directed approach for 129 single parents (98% were mothers) on social assistance in a Canadian setting. A total of 43% of these parents had a major depressive disorder and 38% had two or three other health conditions at baseline. METHODS: Study participants were recruited over a 12 month period and randomized into two groups: one receiving proactive public health nursing and one which did not. RESULTS: At 2 years, 69 single parents with 123 children receiving proactive public health nursing (compared with 60 parents with 91 children who did not receive public health nursing services) showed a slightly greater reduction in dysthymia and slightly higher social adjustment. There was no difference between the public health and control groups in total per parent annual cost of health and support services. However, costs were averted due to a 12% difference in non-use of social assistance in the previous 12 months for parents in the public health nursing group. This translates into an annual cost saving of 240,000 dollars (Canadian) of costs averted within 1 year for every 100 parents. CONCLUSIONS: In the context of a system of national health and social insurance, this study supports the fact that it is no more costly to proactively service this population of parents on social assistance. PMID- 11882102 TI - Cost-effectiveness of ancrod treatment of acute ischaemic stroke: results from the Stroke Treatment with Ancrod Trial (STAT). AB - RATIONALE, AIMS AND OBJECTIVES: This paper describes a recent randomized controlled trial in which 42% of patients receiving ancrod attained a favourable outcome in comparison with 34% of controls. Although the above effect size corresponds to a number needed to treat (to achieve a favourable outcome) of approximately 13, intuition does not necessarily suggest what would be the overall impact of a treatment with this level of efficacy. METHODS: The objective was to evaluate the cost-effectiveness of ancrod. Cost-effectiveness analysis of data from the Stroke Treatment with Ancrod Trial (STAT) trial was carried out. The participants were 495 patients with data on functional status at the conclusion of follow-up. Short-term results were based upon utilization and quality of life observed during the trial; these were merged with expected long term results obtained through simulation using the Stroke Policy Model. The main outcome measure was incremental cost-effectiveness ratio. RESULTS: Ancrod treatment resulted in both better quality-adjusted life expectancy and lower medical costs than placebo as supported by sensitivity analysis. The cost differential was primarily attributable to the long-term implications of ancrod's role in reducing disability. CONCLUSIONS: If ancrod is even modestly effective, it will probably be cost-effective (and, indeed, cost-saving) as well. The net population-level impact of even modestly effective stroke treatments can be substantial. PMID- 11882103 TI - A preliminary study for the development of a scale to assess perceptions about physicians. AB - RATIONALE, AIMS AND OBJECTIVES: This study aimed to develop a scale that can be used to assess the perceptions of the community about physicians and their medical practice. METHODS: A Likert-type scale was developed and applied to 220 subjects twice within a 7-10 day interval. Internal consistency reliability and test-retest reliability were used as indicators of the reliability of the scale. RESULTS: Chronbach's alpha values obtained from two reliability tests were 0.82 and 0.83, respectively, and correlation coefficient between the two tests was 0.61. The mean value of items indicated that perceptions about physicians were between "negative" and "neutral". CONCLUSIONS: The reliability of this scale was found to be at acceptable levels and therefore it can be used to assess the perceptions of people from different social classes about physicians. PMID- 11882104 TI - The rationale of value-laden medicine. AB - Medicine is becoming increasingly confident that scientific advances, especially in the area of genetics, will allow a major improvement in the control and eradication of disease. This development seems to go hand in hand with health enhancement strategies, erasing the distinction between the states of health and disease, and blurring the specific goals of medical services. Medicine tends to become an increasingly technocentric practice that relies heavily on expert knowledge and on epidemiological evidence, neglecting the lived-body experience of being ill, and tending to transform costly medical services into commodities only affordable by the affluent. This paper argues that disease is not merely a functional description, but rather a definitely value-laden organismic state that is experienced by the patient, needs to be explored and treated by medical practitioners, and requires the assessment and participation of social institutions concerned with the delivery and support of medical services. Each of these perspectives introduces its own set of values, both in the clinical encounter and in public health programmes. Bioethics seems to be the appropriate discipline to discuss all these values involved, and help assign them properly in order to rescue the caring concern of medicine for the sick, as well as uphold a principle of fairness in publicly funded medical services. PMID- 11882105 TI - Complaints and claims in the UK National Health Service. PMID- 11882106 TI - Nursing roles and nursing leadership in the New NHS - changing hats, same heads. PMID- 11882107 TI - A collaborative approach to the implementation of clinical supervision. AB - AIM: This paper discusses a collaborative approach to implementing clinical supervision, which was initiated between a primary care trust and a school of nursing and midwifery. BACKGROUND: To enable clinical supervision to proceed successfully and to be perceived as beneficial, this necessitates a collaborative partnership between clinicians, managers and educationalists. KEY ISSUES: The different stages of the initiative will be explored and the paper will consider examples of the collaborative processes involved. The evaluation of the project is examined and suggestions for the future continuation of the initiative are discussed. CONCLUSION: There is evidence that this has been a successful initiative and that a collaborative way of working can be beneficial when implementing clinical supervision. PMID- 11882108 TI - Patient reactions to cancelled or postponed heart operations. AB - OBJECTIVES: The aim was to survey the rate and cause of cancellations of planned cardiac operations at a Swedish clinic during 1999, and to study how the patients were affected. DESIGN: Questionnaires were distributed to 74 patients who had their operations cancelled. Their mood after discharge was measured with The Hospital Anxiety and Depression scale. Ninety-three patients, who were operated on without postponement, served as controls. RESULTS: Sixty-one percent of the patients in the cancellation group reacted negatively, especially if the reason for cancellation was organizational (P = 0.03). The women in the cancellation group had a significantly higher degree of depression than men (P = 0.01) and both women (P = 0.02) and men (P = 0.003) in the control group. Most of the patients, however, were satisfied with the nursing staff's reception and information. CONCLUSIONS: The patients reacted negatively to the cancellation, especially if it had organizational reasons. Women subjected to cancellation had a significantly higher degree of depression than other patients. To be avoided, organizational and medical problems must be identified in time. One way to do this is to introduce a preadmission nurse clinic. PMID- 11882109 TI - Investigating the nursing contribution to commissioning in primary health-care. AB - AIM: This study set out to investigate nurses', doctors' and managers' perceptions of the nursing contribution to commissioning in primary health-care. BACKGROUND: The study was undertaken in the light of the proposal in the government's White Paper The new NHS, Modern, Dependable that community nurses should play a lead role in commissioning health services as members of the new primary care groups. METHOD: Semistructured taped interviews were conducted, with seven community nurses, two community midwives, seven managers and four general practitioners, covering three geographical locations in the UK. The data were transcribed and analysed using a template approach, where text is analysed through the use of an analysis guide, or codebook, consisting of categories, or themes, relevant to the research questions. FINDINGS: The main themes to emerge were that nurses had an important contribution to make to commissioning on the basis of their clinical knowledge, their awareness of health need and their knowledge of health promotion. Additionally, the ability of nurses to negotiate at different levels across professional groups and with the public was perceived as an important skill in a commissioning role. However, findings also indicated that nurses need additional knowledge and skills if they are to perform their role effectively. Being sidelined from the commissioning arena, poor interprofessional collaboration and the potential for resistance from some general practitioners and managers were identified as constraints on the nursing contribution to commissioning. Combining clinical caseloads with commissioning responsibilities and a lack of support systems for nurses in commissioning were also identified as constraints. CONCLUSION: Nurses' commissioning-related knowledge and skills will have to be developed and constraints on the commissioning role of nurses challenged if the perspectives and values of nursing are to help to reshape our healthcare system. PMID- 11882110 TI - Indicators for competent nursing practice. AB - AIMS AND BACKGROUND: This paper identifies and classifies indicators for competent nursing practice and validates these indicators in a variety of settings. Descriptive data to address competent practice in a variety of settings were collected from staff nurses, head nurses and nursing directors in an acute 1000-bed university hospital in Finland. METHOD: The data obtained from 25 expert groups were analysed to identify a clinical set of indicators for competent nursing practice from the data. The relevance of this set of indicators in a variety of clinical settings was further validated with a second sample of expert nurses (n = 26). Thereafter, data were analysed to identify generic competencies that were applicable to all clinical working environments. RESULTS: Twenty-three generic indicators of competent nursing practice were identified in a variety of settings. The findings suggest that these competence indicators are meaningful to nurses with various backgrounds and practice settings. CONCLUSIONS: Collaboration and coordination, as well as the holistic management of the situation, are highly recognized as meaningful characteristics of competent nursing practice. PMID- 11882111 TI - The efficiency of fluid balance charting: an evidence-based management project. AB - This study began following an observation that fluid balance (FB) charts seemed to be overloading medical records in Hong Kong hospitals. The major issues for this study were the efficiency and appropriateness of FB charting. The study design included an analysis of 250 medical records and an opinion survey. Seventy five doctors and 98 nurses participated in the survey. The results indicated that 50% of all medical records had FB charts and in these cases they made up between 12 and 16% of the whole medical record. Intravenous infusion and urinary catheterization accounted for almost 70% of the recorded reasons for using them. Thirty-two per cent of FB charts were found to be incomplete or inaccurate. Forty six per cent of doctors and nurses reported that they believed that charts were not always terminated when they were not required. The most common belief among doctors was that only doctors should discontinue the FB charts. Nurses were almost unanimous in believing that they should not do this without the agreement of the doctors! It was concluded that nurses in the context under study should be empowered to terminate charts when they become unnecessary. In addition, the design of charts should be modified to accommodate different purposes. PMID- 11882114 TI - Exploring the emotional support needs and coping strategies of family carers. AB - This paper explores the emotional support needs and coping strategies of family carers derived from two focus group interviews, each group comprising seven family carers. The interviews were carried out in Northern Ireland as part of phase one of the ACTION (Assisting Carers using Telematics Interventions to meet Older persons Needs) project. The purpose of the interviews was to investigate family carers' needs and experiences, but this paper focuses specifically on the emotional support needs and coping strategies identified by group members. The issues discussed included information and social support needs, the emotional impact of caring, and coping with and adaptation to the caring role. The findings reveal that for the majority of family carers their experience was one of constant searching for support and information. The anxiety and frustration associated with inconsistent and irregular support and lack of information was a major source of concern for the carers in the study. However, the carers employed a number of positive and negative coping strategies to deal with the stress associated with their caregiving role. The study concluded with the recommendation for nurses and other health care professionals to become more proactive in assessing and meeting carers' emotional support needs. PMID- 11882112 TI - Quality of care and development of a critical pathway. AB - BACKGROUND: The current study reports on the first step towards developing quality of care for stroke patients at a hospital in western Sweden. A general critical pathway from the patient's perspective was developed by a multidisciplinary group. AIM: The aim was to investigate quality of care and the effectiveness of a critical pathway. METHOD: The study should be classified as a case study. Patients (n = 22), next of kin (n = 63) and staff (n = 91) make up the research group. Data collection was carried out through questionnaires on quality of care before testing the critical pathway. Data on the critical pathway were collected during the testing period. FINDINGS: The respondents valued some aspects of care as good, but there were some deficiencies as regards patient information and the patient's participation in his/her own care. Next of kin were less positive than the patients. The analyses of the critical pathway showed that the key events need to be more specific and that the number of key events should be reduced. CONCLUSION: By additionally developing the critical pathway, it should be possible to further improve the quality of care for stroke patients. PMID- 11882115 TI - Subjective lack of social support and presence of dependent stressful life events characterize patients suffering from major depression compared with healthy volunteers. AB - The aim of this study was to characterize and compare patients with major depression and healthy volunteers concerning stressful negative life events, social support and social network occurring during a 12-month period. Forty outpatients who fulfilled DSM-IV criteria for major depression and 19 healthy volunteers were included in the study. The results showed no significant differences between the patients and the volunteers concerning the number of independent life events, perceived sufficient persons in the network, the number of family members, having a partner, and perceiving family members as offering the best support. Thus, there were differences in the number of both dependent stressful life events, especially increased arguments with partner and family members, and life events including personal illness. The patient group had experienced a greater number of these events compared with the volunteer group. Comparisons between the two groups revealed differences concerning social support and social network. Fewer patients, especially women, felt they had sufficient social support, had fewer persons in the network, less contact with persons in the network, and fewer confidants compared with the volunteers. PMID- 11882116 TI - From medicalization to hybridization: a postcolonial discourse for psychiatric nurses. AB - I begin with an Orwellian dilemma [Orwell G. (1968) The Collected Essays, Journalism and Letters of George Orwell, Vol. 1, p. 239. Harcourt Brace Jovanovich, New York]: do I 'shoot the elephant' (by writing the abstract) to impress the editor? Or, with the courage of my postmodern convictions, do I lay down my rifle and disregard such suppressive editorial instructions? Bang! My words strafe the paper and the elephant is dead. How difficult it is to stay standing against the powerful currents of the dominant tradition. How easy it is to disavow the inequalities and injustices of that tradition when your livelihood (and your ego) depends upon it. So goes the theme of my paper, that, despite the clarion calls of the illustrious minority to reject the patriarchal model of medical psychiatry, psychiatric nurses continue to be propelled by the twin engines of illness and diagnosis. Yet as soon as psychiatry encounters the 'other' it becomes, in Homi K. Bhabha's words, 'hybridized': a pregnant pause created from the seeds of two different cultures. In this sense, every psychiatric moment becomes a golden opportunity for the psychiatric nurse to abdicate her role as medical factotum. Freed from these contractual obligations, she can join the 'other' and share in his experiences, sustaining rather than negating him within a truly therapeutic alliance. In similar fashion, this article has become a mixture of rhetorical fluidity and structured reality: a hybridized compromise which acknowledges the journal's publication boundaries yet still revels, at times, in the freedom of an open and lyrical text. PMID- 11882117 TI - Protection and advocacy: an ethics practice in mental health. AB - This paper reports the findings of investigations into allegations of patient abuse and the implications for policy and practice. These investigations were carried out by a nurse with a background in ethics for the office of Protection and Advocacy for Individuals with Mental Illness (PAIMI), a state agency operating under a United States federal law entitling it to investigate complaints by psychiatric patients. PAIMI uses investigations both to help individuals and to provide an avenue for broader change. There are four steps in the investigation process: (1) definition of the problem; (2) gathering information; (3) synthesis; and (4) addressing the problem. Cases are presented to illustrate the investigation process and identify ethical issues arising in mental health treatment. Among the issues raised are autonomy and forced treatment, deinstitutionalization, bias against the mentally ill, privacy, and surrogate treatment decisions. Resolutions range from providing individual advice to clients or clinicians, to changes in institutional policy and the publication of guidelines for specific situations. The following lessons were learnt from the investigations: (1) tell patients what to expect; (2) pay attention to the process of giving care; (3) allow patients to feel ambivalent about treatment; and (4) work to develop good relationships; underlying every investigation has been a poor relationship. PMID- 11882119 TI - The relationship between involvement in activities and quality of life for people with severe and enduring mental illness. AB - A number of authors have indicated that people who have greater and more satisfactory social activity tend to report that they have higher life satisfaction. This study was concerned with examining the relationship between involvement in activities and self-reported quality of life for people with severe and enduring mental illness. It is part of a larger study of quality of life for this client group. A random sample of 92 respondents was drawn from the population of people who met the criteria for severe and enduring mental illness in a Health Board in Northern Ireland. Data were gathered using a reliable and valid interview schedule based on a quality of life profile developed for the larger study. The findings indicated that over half of the respondents (56.5%, n=52) had no structured activity in their everyday life and there was a low level of participation in 14 listed activities of living. The respondents also reported a low level of self-reported quality of life. There was also a weak, but statistically significant, correlation between involvement in activities and self reported quality of life. However, a moderate positive statistically significant correlation was found to exist between satisfaction with involvement in activities and self-reported quality of life. The implications for the care of people who are experiencing severe and enduring mental illness within and outside the United Kingdom are discussed. PMID- 11882118 TI - The therapeutic relationship: historical development and contemporary significance. AB - The therapeutic relationship is a concept held by many to be fundamental to the identity of mental health nurses. While the therapeutic relationship was given formal expression in nursing theory in the middle of the last century, its origins can be traced to attendants' interpersonal practices in the asylum era. The dominance of medical understandings of mental distress, and the working-class status of asylum attendants, prevented the development of an account of mental health nursing based on attendants' relationships with asylum inmates. It was left to Peplau and other nursing theorists to describe mental health nursing as a therapeutic relationship in the 1940s and later. Some distinctive features of colonial life in New Zealand suggest that the ideal of the attendant as the embodiment of bourgeoisie values seems particularly unlikely to have been realized in the New Zealand context. However, New Zealand literature from the 20th century shows that the therapeutic relationship, as part of a general development of a therapeutic discourse, came to assume a central place in conceptualizations of mental health nursing. While the therapeutic relationship is not by itself a sufficient basis for professional continuity, it continues to play a fundamental role in mental health nurses' professional identity. The way in which the therapeutic relationship is articulated in the future will determine the meaning of the therapeutic relationship for future generations of mental health nurses. PMID- 11882120 TI - The use of special observations: an audit within a psychiatric unit. AB - Observation is a fundamental skilled nursing intervention. Special observation is an intensified and often prolonged form of this intervention. Depending upon their nature, and the circumstances under which they are carried out, special observations may invoke varying degrees of stress in both the observer and the observed. They may also raise important ethical and significant financial questions. In psychiatry, special observations are usually imposed where a patient is assessed as representing some degree of risk to themselves or others because of their behaviours or potential behaviours. Special observations vary in their degree of intrusiveness and restrictiveness, and may arouse strong emotions in the patient and staff. Patients may be denied privacy for their most intimate needs, and staff may become a focus for patients' acting-out behaviours. Special observations may be medically imposed with minimal consultation with nursing staff, and where there is no nursing and medical staff agreement on their need, nursing staff may at times feel frustrated and powerless to reduce the patients' discomfort with their situation. This paper presents a study into the use of special observations on psychiatric inpatients across a range of clinical settings. The literature on special observations is reviewed, and the findings of an audit into the documentation of special observations is presented and discussed. The study sites comprised two open acute wards, one elderly functionally mentally ill assessment unit which also treats patients with eating disorders, and a secure high dependency unit, which provides rehabilitation within a secure environment for patients with severe and enduring mental illness. PMID- 11882121 TI - Occupational stress and job satisfaction in mental health nursing: focused interventions through evidence-based assessment. AB - In the UK, over 20 contemporaneous reports have shown that between one-quarter and one-half of National Health Service (NHS) staff report significant personal distress (Weinberg & Creed 2000). There exists a substantial body of evidence to suggest that high levels of stress are endemic throughout the NHS (Anderson et al. 1996), and that many of these stressors may be unique to health care (Payne & Firth-Cozens 1987, Calboun & Calboun 1993). Historically, responsibility for stress management has often been placed at the feet of the employee (Sutherland & Cooper, 2000). This individual orientation has lead to the development of practices designed to treat those exhibiting symptoms of stress, and a relative paucity of interventions oriented to the prevention and the management of workplace stressors. It can also serve to deter scrutiny of injurious workplace conditions, practices and procedures. This small-scale study examines stress and job satisfaction in community mental health nurses in a semirural area of North Wales. Based on self-report questionnaire feedback, focused interventions are described to enhance work satisfaction and help ameliorate occupational stressors. Interventions are described at the level of the individual, the team and the organization as a whole. PMID- 11882123 TI - Psychiatric nursing and organizational power: rescuing the hidden dynamic. PMID- 11882122 TI - An integrated model for adolescent inpatient group therapy. AB - This paper proposes an integrated group therapy model to be utilized by psychiatric and mental health nurses; one innovatively designed to meet the therapeutic needs of adolescents admitted to inpatient psychiatric programs. The writers suggest a model of group therapy primarily comprised of interpersonal approaches within a feminist perspective. The proposed group focus is on active therapeutic engagement with adolescents to further interpersonal learning and to critically examine their contextualized lived experiences. Specific client and setting factors relevant to the selection of therapeutic techniques are reviewed. Selected theoretical models of group therapy are critiqued in relation to group therapy with adolescents. This integrated model of group therapy provides a safe and therapeutic forum that enriches clients' personal and interpersonal experiences as well as promotes healthy exploration, change, and empowerment. PMID- 11882124 TI - Something old, something new, something borrowed, something blue. PMID- 11882126 TI - Dismantling formal observation and refocusing nursing activity in acute inpatient psychiatry: a case study. PMID- 11882127 TI - The use of outcome measures to evaluate the efficacy and tolerability of antipsychotic medication: a comparison of Thorn graduate and CPN practice. AB - Assessing the tolerability and efficacy of treatment with antipsychotic medication is a vital part of mental health care. Research has suggested that many side-effects go undetected by clinicians and there is a need to use standardized assessment tools to ensure that treatments are comprehensively evaluated. The training of Community Psychiatric Nurses (CPNs), who provide much of patients' care, should focus on enhancing skills in using such assessments. This study aimed to examine differences in the use of standardized assessments of antipsychotic side-effects and psychopathology by CPNs and Thorn graduates who had received additional training in delivering psychosocial interventions. A questionnaire was sent to 240 Thorn graduates and CPNs practising in England, with an overall adjusted response rate of 54%. Thorn graduates reported using significantly more standardized assessments of side-effects and psychopathology than CPNs. A trend in both groups towards the use of measures that relied on patient self-report of side-effects was observed. This study identified important deficiencies in current CPN practice. A programme of targeted training may be a more realistic and efficient method of enhancing medication management practices in large numbers of CPNs compared to the more expensive and time-consuming Thorn programme. PMID- 11882128 TI - Long-term illness and patterns of medicine taking: are people with schizophrenia a unique group? AB - Patients with schizophrenia relapse because of inadequate levels of medicine taking. Therefore, it seems logical to learn about the factors underpinning their medicine-taking decisions. Further research is urgently needed to explore this area and thereby to refine models of practice, to promote therapeutic interactions with medicine. Existing research tends to share three common deficits. * It is grounded in the worldview of the psychiatrist not the patient. * It studies the influence of symptoms on compliance behaviour in isolation from other potential variables. * It overlooks the potential influence of the chronicity experience in general by examining schizophrenia outwith the context of other enduring illnesses. Overall these deficits may combine to distort the influence of schizophrenic symptoms on medicine decision making and justify a coercive rather than an empowering approach. A pernicious self-fulfilling spiral may be contributing to the problem of relapse in schizophrenia. Potentially the mental health nurse has a role in addressing this problem. PMID- 11882129 TI - Multiprofessional clinical supervision: challenges for mental health nurses. AB - Recent reform and developments in mental health care provision have increasingly espoused the value of multiprofessional teamwork in order to ensure that clients are offered co-ordinated packages of care that draw on the full range of appropriate services available (DoH 1999a; DoH 2000). Supervision in some form is seen as a key part of all professional practice to provide support to practitioners, enhance ongoing learning, and, to a greater or lesser degree, offer some protection to the public (Brown & Bourne 1996, UKCC 1996). Clinical supervision has gained increasing momentum within the nursing profession, but to a large extent this has been within a uni-professional framework -- nurses supervising other nurses. This paper seeks to explore the ways in which multiprofessional working and clinical supervision interlink, and whether supervision across professional boundaries might be desirable, possible, and/or justifiable. Whilst our own view is that multiprofessional supervision is both possible and desirable, we seek to open up a debate, from our perspective as mental health nurses, about some of the issues related to the concept. Our motivation to explore this topic area emanates from our experiences as supervisors to colleagues within multiprofessional teams, as well as the experiences of those attending supervisor training courses. Following a brief overview of the development of clinical supervision in mental health care and recent policy guidelines, some models of clinical supervision are reviewed in terms of their suitability and applicability for multiprofessional working. PMID- 11882130 TI - The value of community mental health nurses based in primary care teams: 'switching the light on in a cellar'. AB - This paper explores the impact of placing Community Mental Health Nurses (CMHNs) at two primary care practices in South Staffordshire. Data were collected by means of a questionnaire which was sent to primary care personnel at these practices, to ascertain their opinions with respect to the contribution of practice-based CMHNs. Overall, primary care personnel were satisfied with the quality of the service received from the CMHNs, especially in terms of improved communication. They felt that the new arrangements enabled a quicker and more efficient access to the services of the CMHN. The results are discussed in terms of the value of having CMHNs within the primary care setting, and in terms of service planning and future recommendations for mental health services within primary care. PMID- 11882131 TI - Implementing research findings into practice: improving and developing services for women with serious and enduring mental health problems. AB - This paper discusses the process that is currently being undertaken to improve and develop services for women with serious and enduring mental health problems in one specialist psychiatric service. The paper begins by highlighting the particular service needs of this group of women. The process of translating the research evidence into practice is then explored by highlighting the comprehensive preparatory work that was required, how a women's special interest group was established, the choice of a model to guide change and development, and a review of progress to date. The paper concludes by arguing that one of the means of maintaining the current momentum for change will be the celebration and acknowledgement of success. PMID- 11882132 TI - The Tidal Model: developing an empowering, person-centred approach to recovery within psychiatric and mental health nursing. AB - Nursing theories and nursing models have a low profile within psychiatric and mental health nursing in the United Kingdom. This paper describes the philosophical and theoretical background of the Tidal Model, which emerged from a 5-year study of the 'need for psychiatric nursing'. The Tidal Model extends and develops some of the traditional assumptions concerning the centrality of interpersonal relations within nursing practice. The model also integrates discrete processes for re-empowering the person who is disempowered by mental distress or psychiatric services or both. The paper reports briefly on the ongoing evaluation of the model in practice. PMID- 11882133 TI - A qualitative study exploring how qualified mental health nurses deal with incidents that conflict with their accountability. AB - In reviewing the literature on accountability within nursing practice, it became clear that accountability is a complex issue. By using a qualitative new paradigm approach, this study explored how 22 mental health nurses deal with the complex issue of conflict in their accountability within their daily practice, by taking a 'snapshot' of their practice. This was achieved by using written critical incidents as proposed by Flanagan (1954), as this allows the researcher to explore the real clinical situations, as identified by the nurse. Cormack's (1983, 1996) technique for analyses of critical incidents was used. The results identified that these nurses are working in an environment in which they have to deal with aggression, suicide and a lack of support from their line managers, as well as from medical colleagues. The nurses in this study identified the importance of team support; clinical supervision and debriefing as methods which help them deal with issues that affect their accountability, within their practice. PMID- 11882134 TI - Nursing students' experience of their first professional encounter with people having mental disorders. AB - The aim of the study was to generate a theoretical model of what nursing students experience in their first professional encounter with people having mental disorders. Data was collected by interviewing 11 nursing students, selected by strategic sampling. Grounded theory analysis was chosen for the study. Findings showed that the students' personal qualities and the patients' behaviour affected the students' experience. When nursing students who had their own needs in focus met patients who rejected them, they felt helpless, and patients wishing to establish a relationship with them confirmed their positive or negative expectations. Nursing students who were focused on patients' needs met the patient as a unique person and felt confident despite being rejected by the patient, as well as proud when the patients wished to establish a relationship with them. Clinical university teachers can use the findings to gain knowledge about how each student feels as well as to guide them through their psychiatric training. The sample is small and further research is needed to generalize the findings of this study and to explore how nursing students, specialist psychiatric nurses and nurses in other disciplines experience the encounter with patients as well as how patients describe encounters with caregivers. PMID- 11882135 TI - Social policy and mental illness in England in the 1990s: violence, moral panic and critical discourse. AB - Violence perpetrated by people experiencing mental illness poses a continuing challenge to practitioners and policy makers in the mental health field. It has been suggested, however, that policy developments in England during the period 1990--2000 became unduly dominated by the perceived need to prevent such violence and in particular that a 'moral panic' occurred following a series of high profile homicides perpetrated by people experiencing mental illness. This paper critically examines the ability of the moral panic theory to offer a cogent explanation of the relationship between media representations, public perceptions and developments in both social policy and legislation during the last decade. Its conclusions, however, suggest that the evidence does not support assertions of a moral panic and that of moral panic theory itself has serious flaws. Ultimately the paper suggests that we must look elsewhere, particularly to Foucault's conceptualization of discourse and to the work of Birkland, an American political theorist, in our search for an understanding of the relationship between events, ideas and social policy. PMID- 11882136 TI - Knowing me knowing you: towards a new relational politics in 21st century mental health nursing. PMID- 11882137 TI - Being positive about schizophrenia. PMID- 11882138 TI - Commentary on 'Being positive about schizophrenia'. PMID- 11882139 TI - A review of clinical risk and related assessments in forensic psychiatric units. PMID- 11882141 TI - Women in special hospitals: understanding the presenting behaviour of women diagnosed with borderline personality disorder. AB - This paper critically examines the development of the concept of borderline personality disorder (BPD) in terms of the assumed centrality of abnormal early environments and abusive relationships. It is suggested that if BPD is conceptualized as an expression of past experiences in adult life, information regarding early histories can assist in 'making sense' of later behaviour. The aim of this review therefore is to explore how histories of women diagnosed as BPD, within a High Secure Psychiatric Hospital, may facilitate an interpretation of the 'adaptive' nature of presenting 'symptomology'. Case note material is utilized to gain insight into specific aspects of childhood experiences that have been documented, and are thus deemed significant. These findings support the perception that the role of the early environment and associated relationships are significant within written accounts of women diagnosed as having BPD. By exploring the links between trauma and BPD, this article suggests that an understanding of the effects of trauma and the importance of relationships can offer a way forward for self-reflection and future care. PMID- 11882143 TI - Critical incident stress debriefing following traumatic life experiences. AB - This study was designed to explore the use of critical incident stress debriefing as a therapeutic intervention following traumatic life events. A case study approach was used to allow the researchers to adopt a more flexible and overtly involved stance. Initial contact took place 24 h following the traumatic life experiences of three women. Critical incident stress debriefing was provided and data were collected and recorded within an ethical framework. Six months following the traumatic life experiences the women were interviewed again to explore their perceptions of the intervention that was provided. The results demonstrated positive outcomes. The women concluded that the debriefing intervention provided a safe forum for them to explore their needs, process their experiences and create constructive narratives. A carefully constructed critical incident stress debriefing intervention was used within the context of its objectives and acknowledged limitations. The study was small and generalizations cannot be made to other individuals who experience similar tragedies. Nevertheless, evidence from previous research coupled with the findings from this study suggests that mental health nurses might benefit from being educated and trained in critical incident stress debriefing. Further research needs to be carried out to explore the use of different models of stress debriefing applied to special circumstances. The goal of such interventions should be to alleviate symptoms and to prevent the development of a full-blown post-traumatic stress disorder. PMID- 11882142 TI - Evidence-based care and the case for intuition and tacit knowledge in clinical assessment and decision making in mental health nursing practice: an empirical contribution to the debate. AB - This paper provides empirical evidence that challenges the view that methods of clinical assessment and decision making should not rely solely on logical positivist approaches. Whilst the National Health Service (NHS) Executive currently takes a hard positivist line on what constitutes evidence-based practice, data reveal that it is not always appropriate to disregard the tacit knowledge and intuition of experienced practitioners when making assessment decisions in mental health nursing practice. Data support the case for a holistic approach which may draw on intuition and tacit knowledge, as well as traditional approaches, to meet the requirements of clients with complex mental health problems. A model based on Schon's notion of reflection in and reflection on practice is proposed which demonstrates the value of intuition and tacit knowledge. This model allows the generation of insights which may ultimately be demonstrated to be acceptable and empirically testable. It is accepted that an element of risk taking is inevitable, but the inclusion of a formal analytical process into the model reduces the likelihood of inappropriate care interventions. The cognitive processes which experienced nurses use to make clinical decisions and their implications for practice will be explored. PMID- 11882144 TI - Mass media, 'monsters' and mental health clients: the need for increased lobbying. AB - A review of the limited empirical and theoretical literature indicates that current mass media representations of mental health service users appear to emphasize violence, dangerousness and criminality. This is despite the empirical evidence that indicates a decline over the last 40 years in the number of homicides carried out by people identified as suffering from mental health problems. Such inappropriate representations do much to increase stigma, ostracism, harassment and victimization of these individuals by the public. Furthermore, it can be argued that there is another repercussion of these representations and that is the subsequent government position/policy and the resulting legislation concerning care of people with mental health problems. Consequently, this paper argues that there is a clear need for psychiatric/mental health (P/MH) nurses to become more mindful of the wider, socio-political environment in which their practice occurs, particularly if psycho-social approaches to practice are adopted in their fullest sense, and as a result increase their political lobby. Such increased lobbying should occur on behalf of, and in collaboration with, service users, and accordingly the authors describe a range of activities under the broad headings of pro-active and reactive lobbying. Furthermore, it is incumbent upon P/MH nurse educationalists to prepare aspirant P/MH nurses for this lobbying role and equip them with the skills necessary to do so. PMID- 11882145 TI - Interventions in the nurse-patient relationship in forensic psychiatric nursing care: a Swedish survey. AB - Nurses (Registered nurses, RN, and Licensed Mental Nurses, LMN) working in five Swedish forensic psychiatric units filled in a questionnaire designed for general psychiatric nursing, but modified for forensic use. In this report, data concerning the extent to which treatment interventions included verbal elements, and the focus of these nurse--patient verbal interactions, were analysed. The most commonly used interventions were 'social interaction', 'regular communication' and 'social skills training'. The most common focus in verbal nurse--patient interaction were 'explaining consequences, confronting and encouraging the patient to talk about his/her crime/behaviour', 'interpretative communications with the patient', and 'communication about functions in daily life'. The salient findings were: (1) seven of the fifteen interventions used by the nurses included verbal elements to a great extent; (2) nurses often used confronting interventions, despite the fact that the large EE (expressed emotion) literature suggest that hostility and critical remarks may provoke a worsening of symptoms among psychotic patients; (3) there was only little correspondence between actual practice and theoretical models; and (4) there were unexpectedly small differences between the two professional groups (RN and LMN), suggesting that the roles are not distinct. PMID- 11882146 TI - Seeking a clarification of meaning: a phenomenological interpretation of the craft of mental health nursing. AB - This paper gives a description of mental health nursing practice provided by nurses working in a community mental health resource centre, in an inner city area of a northern English city. The centre offered a mental health service for a mixed client group who had enduring and/or acute mental health problems. The nurses engaged as the subjects in a phenomenological study of the nature and meaning of their practice in order to improve the care they gave the clients. The findings from the study suggest that, for the nurses, the meaning of nursing was characterized by holism, relationship-building, partnership and empowerment. The findings revealed that the aim of nursing was to promote a positive self-concept in clients and this was achieved by exploring the lived experience of the client and the nurse through relationship-building processes. PMID- 11882147 TI - The aetiology, presentation and treatment of personality disorders. AB - Personality disorders are a heterogeneous collection of conditions with common features, which may include an exaggerated self-centred nature, little regard for the feelings of others, or the regular fabrication of stories to explain the behaviour of self or others. Whilst such features might be recognized as being present in many people at different times of their lives, it is the persisting nature and extremes of personality traits that distinguishes those who have personality disorders from those who do not. Apart from the problems that personality disorders bring on their own (such as dysfunctional relationships), when they coexist with mental disorders it makes the latter more difficult to treat. People with personality disorders are often depicted as being dangerous, yet only a few are and it is this minority group that attract public attention. Personality disorders are recognized as belonging to the group known as the serious mental illnesses, a group that mental health nurses are being encouraged to focus their attentions on, but it is accepted that there is a paucity of education and training in appropriate interventions for this group of people. This article provides an overview of the aetiology and presentation of personality disorders together with an examination of evidence-based therapeutic interventions. PMID- 11882148 TI - A plea for respect: involuntarily hospitalized psychiatric patients' narratives about being subjected to coercion. AB - Eighteen involuntarily hospitalized psychiatric patients narrated their experiences of being subjected to coercion and their thoughts on how to prevent the coercion. A qualitative content analysis identified recurring themes, which were incorporated in two core themes describing the participants' experience. The core theme Not being respected as a human being included most of their narrated experiences, described in the themes Not being involved in one's own care, Receiving care perceived as meaningless and not good, and Being an inferior kind of human being. The core theme Being respected as a human being included a minor part of the narrated experiences and how the participants wanted things to be, described in the themes Being involved in one's own care, Receiving good care, and Being a human being like other people. The participants' plea for respect is discussed in relation to the ongoing deinstitutionalization of psychiatric care and the need for attitude changes in care and community, leading to the treating of mentally disordered people with more respect. PMID- 11882149 TI - Absurdity and being-in-itself. The third phase of phenomenology: Jean-Paul Sartre and existential psychoanalysis. AB - Existentialism and phenomenology are closely linked philosophies. Existentialism preceded phenomenology and is not considered a single philosophy but several schools of thought, both theist and atheist in thinking, which grew out of a reaction to traditional philosophy. The development of phenomenology is divided into three separate phases ultimately merging with existentialism. Following Second World War, the phenomenological movement gained momentum in France and encompassed many of the ideas of Edmund Husserl and Martin Heidegger. Gabriel Marcel, Maurice Merlieu-Ponty and, notably, Jean-Paul Sartre established a 'third phase' of phenomenology. This paper explores some of Sartre's ideas related to being and later applications through Medard Boss and R.D. Laing, and offers a short illustrative case vignette that shows the concepts as they might apply to nursing practice. Consideration is finally given to existential psychoanalysis as an applied research methodology PMID- 11882156 TI - Biomedicalization and changes to professional education: two factors in the psychiatric nursing "identity crisis". PMID- 11882157 TI - Mental health nurses' PRN psychotropic medication administration practices. AB - PRN psychotropic medications (medications given with the aim of changing the patient's mental state on an 'as needed' basis) are frequently prescribed and administered in mental health facilities. Their administration is largely an autonomous nursing role. However, little research has explored this important area of nursing practice in any detail and therefore the use of this intervention by mental health nurses is not fully understood. A descriptive project was undertaken to explore the factors involved in the administration of PRN psychotropic medications. It utilized a retrospective chart audit, conducted at one regional and one urban setting in Queensland, Australia. The results indicate a lack of clarity surrounding psychotropic PRN medication administration practices, confusion surrounding decision-making processes related to this intervention, and poor documentation practices. The study found a high incidence of PRN psychotropic medication administration when compared to similar studies, and an excessive reliance on the use of typical antipsychotics over benzodiazepines. This practice is not supported by contemporary literature. As a result of our findings, we recommend further studies in areas of related importance, in order to increase our understanding of the complexities surrounding this important nursing intervention, and an investigation into documentation practices in inpatient mental health settings. PMID- 11882158 TI - In harm's way: a postmodern narrative inquiry. AB - Deliberate self-harm is a traumatic experience which has serious consequences at individual and social levels. Differing motivations for self-harm and different modes of injury require that therapeutic responses vary. Yet, according to the literature and personal observation, nursing responses tend to be uniform and inflexible. Not only are such responses inadequate for effectively dealing with individuals who self-harm, they also have unintended, hidden deleterious consequences for nurses and nursing. In this paper, a story about self-harm is analysed using a postmodern perspective that helps to illuminate multiple dimensions to this complex issue. The approach enriches understanding, opening up possibilities to improve the quality of the nurse--patient relationship. PMID- 11882159 TI - Exploring the relationship between work stress, years of experience and emotional competency using a sample of Australian mental health nurses. AB - This paper reports an exploratory study examining the relationship between emotional competency and work stress, and the association between the length of time in mental health nursing and emotional competency and stress. A significant relationship was found between emotional competency and years of experience. Nurses with six years and more experience had higher levels of emotional competency. This relationship was stronger for female than male nurses. Nurses with less than two years in the nursing profession were found to experience significantly more personal self-doubt than nurses with greater nursing experience. PMID- 11882160 TI - HoNOS: is there any point in training clinicians? AB - This study examines the impact of training and clinical experience on the inter rater reliability of the HoNOS. Following either standard or modified HoNOS training, two groups of registered mental health nurses (RMHNs) were tested for inter-rater reliability (IRR) using a standard case vignette. In addition, another group of nurses and two nonclinical comparison groups had their IRR tested using the same vignette but without receiving any HoNOS training. There was no significant difference between IRR scores post-training compared with the pre-training scores. This is the case whether the recommended or modified training programmes were used. In addition, there is no significant difference between nurses and the comparison group, characterized by nonclinical work experience in a mental health setting. All four groups achieved adequate IRR. Finally, the comparison group with no regular exposure to mental health patients but experience using rating scales are significantly less reliable than the other four groups. The results suggest two possible interpretations. The first is that the HoNOS scales are so well designed that only familiarity with psychiatric patients and no experience with rating scales is sufficient to achieve adequate IRR even without training. Alternatively, the use of written vignettes does not provide a valid measure of HoNOS IRR. PMID- 11882161 TI - An overview of the role and functions of a psychiatric consultation liaison nurse: an Australian perspective. AB - The role of the psychiatric consultation liaison nurse (PCLN) has increased substantially in popularity over the last few years. Despite the growth of this position, a paucity of literature regarding the role, functions and effectiveness of psychiatric consultation liaison nursing continues to exist. The current study was undertaken as part of the Victorian Nurse Practitioner Project. A significant aspect of this study concerned collection of data on the activities of the PCLN. This approach enabled an extensive and detailed profile of the PCLN to be formulated. The findings indicate that the PCLN provided a service to nursing, medicine and allied health in relation to patients experiencing mental health problems in the general hospital setting. Patients referred to the PCLN presented varied clinical features in terms of medical, surgical and mental health disorders. The PCLN performed a range of interventions. The results of this study make a significant contribution to address the current paucity of literature. PMID- 11882162 TI - Mental health professionals' attitudes towards people who have experienced a mental health disorder. AB - The aim of this study was to gain information about the attitudes of mental health professionals towards people who have experienced a mental illness. The study involved a survey of 266 mental health professionals employed in a range of mental health treatment settings. Respondents were administered a questionnaire based on one of two vignettes describing a person with schizophrenia or depression. Attitudes were represented as beliefs about prognosis and long-term outcomes in relation to the two vignettes. Professional groups were found to be less optimistic about prognosis, and less positive about likely long-term outcomes, when compared with the general public. Medical staff were less optimistic about outcomes than other professional groups, with mental health nurses generally most optimistic. Most professionals based their attitudes on their experiences of working with people with mental health problems. PMID- 11882164 TI - Bipolar disorders: an overview of current literature. AB - This paper aims to review current literature around bipolar disorder from a multidisciplinary stance. As a result of a literature search, several key themes have been identified which will be discussed during this paper: first, the definition of bipolar disorders, their epidemiology and the effect of the illness on outcome, and secondly pharmacological interventions, psychosocial interventions and the use of coping strategy enhancement for this client group. The paper attempts to direct clinicians towards relevant further reading and research for the development of interventions when working with this client group. PMID- 11882163 TI - What are the principles and processes of inspiring hope in cognitively impaired older adults within a continuing care environment? AB - This study focuses on the principles and processes that psychiatric/mental health (P/MH) nurses use to inspire hope in cognitively impaired older adults within a National Health Service (NHS) continuing care environment. Using a grounded theory method, the data were coded and analysed, in an attempt to produce an integrated conceptual framework of hope inspiration. This comprised four core variables: applied humanistic code, pragmatic knowledge, interpersonal relations and nurse as utilizer. The authors postulate a relationship between the function of caring, the activity of helping and the practice of instilling hope, as a basis for suggesting that inspiring hope to clients is one of the primary acts of P/MH nursing. The authors conclude that the four core variables are intertwined and inseparable from one another, and bound up with P/MH nursing practice. From this position, it is suggested that the processes of inspiring and instilling hope cannot be separated from the qualities of being a P/MH nurse. However, a more detailed and complete understanding of the concept of hope can increase the nurse's effectiveness as a carer. PMID- 11882165 TI - Person-centred care for people with dementia: a quality audit approach. AB - This paper addresses the concept of person-centred care for people with dementia by consideration of an audit process using dementia care mapping as the audit tool. It is argued that this tool is best for identifying the lived experiences of the people in receipt of care. As a result it is able to identify the overall culture of care and its level of 'person-centred' approach. The audit was conducted on 12 units, half of which were day units and the others catering for inpatients. Five patients were mapped on each day for a 4-day period. The results give some idea of the quality of care and identify where improvement is necessary. Scores such as well-being values and the Dementia Care Index give clear signposts to the level of person-centred care and highlight where staff development is necessary. Recommendations are given to aid on-going planning. PMID- 11882166 TI - Developing A&E nursing responses to people who deliberately self-harm: the provision and evaluation of a series of reflective workshops. AB - Accident & Emergency (A&E) departments and Medical Admission Units (MAUs) are an essential part of emergency mental health care in the UK. Deliberate self-harm and attempted suicide are particular challenges to nursing staff in such departments. Two registered mental nurses with experience of education facilitated a series of workshops with nurses from four A&E departments, two Minor Injuries Units and two MAUs. These were focused upon the assessment and management of deliberate self-harm and attempted suicide. During the workshops, participants provided systematic information on presentations of deliberate self harm and attempted suicide, and on their professional responses to such presentations. These accounts provided invaluable information on deliberate self harm and attempted suicide as seen by nursing staff in A&E and MAUs. Critical reflection upon nursing responses to these presentations revealed particular concerns nurses had relating to their experience with this client group which might inform future practice. Evaluation of the workshops indicated a subsequent decrease in work-related stress, which was probably a consequence of improved professional coping responses by the nurses concerned. PMID- 11882167 TI - Addiction and creativity: from laudanum to recreational drugs. PMID- 11882168 TI - Rejoinder to Barker and Clarke. PMID- 11882169 TI - Doubts and certainties in the nursing profession: a commentary. PMID- 11882170 TI - Psychometric development of the Mental Health Problems Perception Questionnaire. PMID- 11882171 TI - Sternal Wound Infection Prediction Scale: a test of the reliability and validity. AB - The purpose of the present study was to determine the reliability and validity of the Sternal Wound Infection Prediction Scale. The tool was developed to fill the need for a quick and easy way to determine which patients are at risk of developing a sternal wound infection in order that preventive measures can be instituted. Data were collected by a retrospective chart review of 56 patient records, 27 who developed a sternal wound infection and 29 who did not. In this group of patients, obesity and diabetes were found to be statistically significant risk factors. Pharmacological support was found to be a clinically significant factor. Of the infected group, 68.4% of patients who developed a sternal wound infection were predicted to do so by the tool. PMID- 11882172 TI - Self-esteem and student nurses: an account of a descriptive study. AB - In this paper the authors describe a study of nursing students' self-reported self-esteem levels using a validated instrument: the Culture-Free Self-Esteem Inventory 2 (CFSEI-2). The research question was: How do student nurses rate their own self-esteem levels? The paper offers a short review of some of the literature, followed by a description of the sample (a convenience sample of 101 undergraduate nursing students), data collection and analysis methods and the findings. The self-esteem components of CFSEI-2 are general, personal and social and these terms are defined in the text of this paper. The findings indicated that the mean scores for this sample of nursing students fell within normal levels for all three components of self-esteem. Younger students were found to have higher scores for the social subscale, which indicates that they had a higher perception of the quality of their relationships with their peers. The findings of this study indicate that these undergraduate student nurses' perceptions of their self-esteem were comparable to the normal ranges of self esteem as assessed by the instrument. PMID- 11882173 TI - Using Internet services to generate a research sampling frame. AB - The purpose of this paper is to describe how the authors used Internet services to generate a sampling frame for their on-going pilot study, which evaluates the psychometric properties of Chen's Nursing Home Tool, a community needs assessment instrument for nursing homes. The proposed sampling frame may be used to survey nursing home residents and family members, nursing home providers and community leaders for any selected community in the USA. The availability of a user friendly sampling frame can assist health-care professionals to determine required and desired services and further contribute to the development of a more comprehensive health-service system. PMID- 11882174 TI - Complementary therapy use by nursing, pharmacy and biomedical science students. AB - Attitudes towards the use of complementary therapies by students of undergraduate Bachelor of Nursing, Pharmacy and Biomedical Sciences were determined using a self-administered questionnaire. Overall, 78% of students had used a complementary therapy in the past year and 56% had visited a complementary therapy practitioner. The therapies most used were those involving vitamins, mineral and other supplements. Practitioners specializing in this area were the most visited, followed by chiropractors. Commonly used products included vitamin C, multivitamins, B group vitamins, garlic, iron and echinacea. Most students thought complementary therapies improved quality of life, with friends and family providing the main sources of information. There were few differences attributable to course or gender. The results suggest that these students have favorable attitudes towards complementary therapies and that many choose to use them as part of normal health care. PMID- 11882175 TI - Nurses' use of restraints and their attitudes toward restraint use and the elderly in an acute care setting. AB - A descriptive correlation study was conducted in an acute-care hospital to explore the relationship between nurses' use of restraints and their attitudes toward restraint use and the elderly. A total of 201 nurses returned a questionnaire that collected demographic information and included two research instruments: (i) Perceptions of Restraint Use Questionnaire and (ii) Attitudes toward the Aged Semantic Differential. Results showed slightly positive attitudes towards the elderly and toward the use of restraints, although there was no correlation between scores on the two scales. Furthermore, nurses' attitudes did not predict their self-reported use of restraints. PMID- 11882176 TI - Improving pain management by nurses: A pilot Peer Intervention Program. AB - Significant pain continues to be reported by many hospitalized patients despite the numerous and varied educational programs developed and implemented to improve pain management. A theoretically based Peer Intervention Program was designed from a predictive model to address nurses' beliefs, attitudes, subjective norms, self-efficacy, perceived control and intentions in the management of pain with p.r.n. (as required) narcotic analgesia. The pilot study of this program utilized a quasi-experimental pre-post test design with a patient intervention, nurse and patient intervention and control conditions consisting of 24, 18 and 19 nurses, respectively. One week after the intervention, significant differences were found between the nurse and patient condition and the two other conditions in beliefs, self-efficacy, perceived control, positive trend in attitudes, subjective norms and intentions. The most positive aspects of the program were supportive interactive discussions with peers and an awareness and understanding of beliefs and attitudes and their roles in behavior. PMID- 11882177 TI - Professional identity of Japanese nurses: bonding into nursing. AB - The purpose of this study was to explore the process of establishing the professional identity of Japanese nurses. Following a grounded theory design, data were generated by interviews, multisite participant observations and theoretical memos. Eighteen Japanese nurses who were selected by theoretical sampling were formally interviewed. Data were analyzed using methods of constant comparative analysis. Six categories emerged from the data: (i) learning from working experiences; (ii) recognizing the value of nursing; (iii) establishing one's own philosophy of nursing; (iv) gaining influence from education; (v) having a commitment to nursing and (vi) integrating a nurse into self. The core category, 'bonding into nursing', incorporated the relationship between and among all categories and explained the process of establishing the professional identity of Japanese nurses. 'Bonding into nursing' was described as an initial substantive theory, which is defined as the process by which each nurse established her/his professional identity as a nurse. PMID- 11882178 TI - Use of nursing homes in Baccalaureate nursing education. AB - Although the use of nursing homes in nursing education has been described in the published research, no current report exists outlining guidelines for faculty if they desire to use nursing homes for their undergraduate nursing courses. Thus, the purpose of this paper is to identify courses that are best taught in nursing homes and the approaches faculty can implement when using nursing homes for student experience. The use of nursing homes for the courses in fundamentals, leadership and management, research and community health is discussed. A step-by step approach for faculty to initiate the use of nursing homes for undergraduate teaching is also provided. The nature of the faculty, the characteristics of nursing home settings and the characteristics within the community will affect students' learning experiences and effectiveness. Thus, it is important to consider nursing homes as appropriate for certain types of learning needs. PMID- 11882180 TI - Burnout versus depression and sense of coherence: study of Greek nursing staff. AB - We assessed the relationships between burn out, depression and sense of coherence (SOC) using the Maslach Burnout Inventory, Beck's depression inventory and Antonovsky's SOC questionnaire in a large sample of 17 male and 62 female Greek nurses. Analysis showed that SOC in nursing staff was correlated with burnout (BO) and depression, while the latter was correlated to a lesser degree with BO. The relationship of BO with depression seems to be the result of the relationship between depression and SOC. Thus, we could hypothesize that the degree of SOC renders persons either vulnerable or resistant to both depression and BO, though further studies are warranted. In the hospital setting, the intervention by specialized personnel in order to assist nursing staff with BO may be an appealing option. Further, prevention programs aimed at helping individuals prone to BO could also be envisaged. PMID- 11882179 TI - Evaluation of the Thai-Lao Collaborating Nursing Manpower Development Project using the Context Input Process Product model. AB - The purpose of this article was to describe, monitor and evaluate the Thai-Lao Collaborating Nursing Manpower Development Project using Stufflebeam's Context Input Process Product model. The study found that the project had great success in personnel development according to the six activities of the project. It helped develop two nurses with Masters degrees, 24 senior nurses and 283 diploma and auxiliary nurses. There were visible changes in the discipline and in terms of nursing care, nursing process was utilized. The Lao nursing system was developed to the point where white uniforms were used as the symbol of nurses who previously went unrecognized. There was a change in the structure of nursing organization. Moreover, nursing activities moved towards the goal of health promotion and prevention. The study recommended that a periodic supervisory activity be implemented to enhance the training of trainers. In addition, a 2 year continuation program for Bachelor degrees in nursing should be developed initially in Laos, then a 4-year program should be developed. The nursing education program for Masters degrees should initially be obtained through neighboring countries. PMID- 11882181 TI - Australian hospital generalist and critical care nurses' perceptions of doctor nurse collaboration. AB - Previous researchers have indicated that collaborative practice between doctors and nurses results in positive effects on patient care, health-care costs and provider satisfaction. Despite these benefits, collaborative practice appears to be the exception, rather than the dominant pattern, within health care. A collaborative relationship cannot evolve if individuals do not value and respect others' competencies. This study, a mailed survey, used the Collaboration with Medical Staff Scale to compare the perceptions of doctor-nurse collaboration held by critical care nurses and generalist hospital nurses. The hypothesis that critical care nurses perceive there to be greater collaboration with doctors than their generalist nurse colleagues was supported even after taking into consideration education and experience. These results suggest that critical care is an area that might be useful when trying to understand the dimensions and implications of collaboration among health professionals. PMID- 11882182 TI - Factors influencing job satisfaction and ethical dilemmas in acute psychiatric care. AB - This study addressed the factors that nursing staff perceived as creating job satisfaction in their working environment in addition to addressing the ethical dilemmas that staff experienced within an acute psychiatric care setting. It also addressed how clinical supervision contributed to job satisfaction among staff as well as the differences between staff who attended and staff who did not attend to clinical supervision. Data were analyzed using descriptive statistics. Overall, the results of this study showed that the factors influencing nurses are related to areas of dissatisfaction, for example, stress and experiences with shortcomings. Factors that contribute to job satisfaction or dissatisfaction were found to be related to the nurses' value systems. The ethical dilemmas that were specifically addressed involved how to care for patients and handle work in relation to patients' autonomy, how to approach the patient, how to provide care against the will of the patient, and what action was ethically right for each particular patient. PMID- 11882183 TI - Hyperglycemia and its related factors in Taiwanese middle-aged adults. AB - Our study used data collected in Chung-Hsing Village to evaluate the relationship between hyperglycemia and its related factors in Taiwanese middle-aged adults in May 1998. All government employees at the Taiwan Provincial Government in Chung Hsing Village aged 40-64 years were candidates for this study. Only 708 volunteers completed a structured questionnaire and underwent venous blood tests. To study the significant related factors of hyperglycemia, the t-test, chi(2) analysis and multivariate logistic regression were used. Our results showed that 40% of the 708 subjects were men and 60% were women. The mean age was 50.5 plus minus 6.8 years. The mean values of fasting glucose were 4.68 plus minus 1.92 mmol/L in men and 4.70 plus minus 1.70 mmol/L in women (P > 0.05). The proportions of hyperglycemia were 6.4% in men and 5.7% in women (P > 0.05). After controlling for other covariates, the multivariate logistic regression analysis showed that the significant related factors for hyperglycemia were older age (odds ratio=2.86, 95%; confidence interval=1.36-6.05; P < 0.01) and hypertriglyceridemia (odds ratio=3.34, 95%; confidence interval=1.68-6.64; P < 0.001). PMID- 11882184 TI - Changes in the regression slope correlating between urinary contents of alpha-1 microglobulin and ulinastatin and its relation to severity in mood disorders. AB - With mood disorders, clinical portrayals alone do not always reflect the exact state and progress of the disease. The present study attempted to evaluate whether changes in regression slope correlating between urinary contents of alpha 1-microglobulin and ulinastatin provided objective information on the severity of mood disorders, based on our previous findings that the regression slope was more steeply inclined in patients with mood disorders than in age-matched healthy subjects. As a result, a close association between scores of the Hamilton Rating Scale for Depression and the regression slopes was found: the more inclined the slope, the greater the severity of symptoms of depression. These results suggest that changes in the slope of the regression plot correlating between urinary contents of alpha 1 M and UT may afford a useful objective index when monitoring the state of patients with mood disorders. PMID- 11882185 TI - Community nurses: an urgent need. AB - When families could not manage to care for the sick, they turned to people who could provide home nursing care. Gradually, however, the sick were moved to institutions and nursing became institution-based. This trend accelerated in almost all countries around the world. Today we see that nurses participate in curative functions, and are concerned with uncommon and/or complex diseases. Very few nurses work in the community to provide services such as illness prevention, encouraging and supporting communities and individuals to participate in their own health care, health promotion and maintenance, and assisting with rehabilitation that address the health needs outside of institutions aimed towards achieving an optimal quality of life. PMID- 11882186 TI - Characteristics of housebound elderly by mobility level in Japan. AB - The purpose of the present study was to describe characteristics of the housebound elderly by their mobility levels. Disabled elderly people in Japan were interviewed at home by nurses. Housebound status was defined as people who left the house less than once a week. Characteristics of housebound elderly were compared with non-housebound elderly by three mobility levels: (i) Non-walking group (n=72); 5 m-walking group (n=153); going-by-bus group (n=96). Housebound elderly were not found in the going-by-bus group. Among the elderly in the non walking group, the cognitive function of the housebound elderly was significantly lower and the housebound elderly were less well cared for; however, their caregivers rated their caregiving burden significantly lower than did caregivers of the non-housebound elderly. Among those in the 5 m-walking group, the autonomy and social networks of the housebound elderly were significantly less than for the non-housebound. The characteristics of housebound elderly showed differences by mobility levels. PMID- 11882187 TI - Challenges of measuring and linking patient outcomes to nursing interventions in acute care settings. AB - Measuring and linking patient outcomes to nursing intervention is an important task that has professional, financial and political ramifications. The importance and complexity of measuring patient outcomes accurately should not be overlooked, as there are a number of emergent factors that influence this process. These include the turbulent context of practise, variations in care due to the large number of health professionals, individual patient characteristics impacting on outcomes, determining appropriate nursing outcome measures, nursing's lack of autonomy within the system and difficulties experienced while trying to link patient outcomes to nursing interventions. So that the results reflect reality, it is important for researchers in the field to take note and consider these factors when measuring patient outcomes. The present article aimed to examine and discuss a number of these factors as they relate to the evaluation of patient care. PMID- 11882188 TI - Fever management practises: what pediatric nurses say. AB - Pediatric nurses manage fevers in hospitalized children daily: a complex practise. The present study identified varied decision-making criteria and inconsistent practise influenced by many external variables. Nurses perform comprehensive assessments in order to make informed decisions. However, factors influencing their practise include medical orders, the temperament of the child, a history of febrile convulsions, parental requests, colleagues and ward norms. Nurses have a 'temperature' at which they consider a child febrile (37.2-39.0 degrees C) and many reported a 'temperature' at which they administered antipyretics (37.5-39.0 degrees C). Antipyretics were administered to febrile children for pain relief, irritability, at the request of parents and to settle a child for the night. Administration was reported to be higher during the day and evening shifts, at medication rounds and when the ward was busy. At night, nurses were reluctant to wake a sleeping febrile child, preferring to observe them instead. Recommendations to promote consistent fever management practises are included. PMID- 11882189 TI - Study on the correlation between depression and quality of life for Korean women. AB - Korean women have many experiences of the negative impacts caused by changes in biological, psychological and socioeconomic environments. Therefore, they often have unique health problems that require specific attention. In particular, depression is a health problem that severely impacts on quality of life for women. The purpose of this study is to investigate the correlation between depression and quality of life for Korean women. The subjects of the study were 474 women (20-60 years) living in Seoul and were chosen by convenient sampling from 12 March to 18 April 2000. The data were collected by a structured questionnaire that included general characteristics, the Center for Epidemiologic Studies Depression Scale and the SmithKline Beecham 'Quality of Life' Scale. The data were analyzed by the SAS 6.12 program, which included descriptive statistics: t-test, ANOVA, and Pearson correlation coefficient. The mean score of depression was 18.5 (range: 4-52), and the mean score of the quality of life was 153.7 (range: 65-230). In the general characteristics, there was a statistically significant difference in the degree of depression according to education (F=5.62, P=0.0009), marital status (F=9.20, P=0.0001), family structure (F=3.97, P=0.0036), level of income (F=10.79, P=0.0001), type of job (F=2.99, P=0.0191), stressors in the job (F=2.56, P=0.0198) and sexual activity (F= -2.56, P=0.0109). As for the general characteristics, there was a statistically significant difference in the degree of the total score for quality of life according to education (F=5.24, P=0.0015), level of income (F=11.15, P=0.0001), employment status (F=2.23, P=0.0268), job satisfaction (F=4.42, P=0.0155) and stressors in the job (F=2.62, P=0.0174). Significantly, there was a negative relationship between depression and quality of life (gamma= -0.5984, P=0.0001). The most significant negative relationship among the five factors of quality of life was between depression and the psychological well-being factor (gamma= -0.5708, P=0.0001). In conclusion, the results of this study will contribute to the development of a nursing intervention program for decreasing depression, preventing depression and promoting quality of life for Korean women. PMID- 11882190 TI - Maternal expectations of the child health nurse. AB - The main aim of this study was to identify what first-time mothers in Sweden expected of Child Health Nurses. A further aim was to investigate what help and support was received by the new mothers and whether they felt that anything was missing. Twenty new mothers were interviewed according to grounded theory and the data were analyzed by the constant comparative method. It was found that first time mothers expected Child Health Nurses to have faith in a new mother's own strength and to be accessible, approachable and knowledgeable, providing advice and support. New mothers indicated that they had experienced most of these characteristics, especially accessibility and approachability, but some felt that there were deficiencies (e.g. a lack of continued interest in their own bodies and health, as well as a lack of support when they ceased to breast-feed). The Child Health Service was taken for granted by new mothers. First-time mothers indicated that they appreciated the service, particularly those mothers who did not have a good social network. However, some of the new mothers expressed negative feelings regarding the Child Health Service provided. PMID- 11882191 TI - Determinants of nurses' intention to administer opioids for pain relief. AB - A statewide cross-sectional survey was conducted in Australia to identify the determinants of registered nurses' intention to administer opioids to patients with pain. Attitudes, subjective norms and perceived control, the key determinants of the Theory of Planned Behavior, were found to independently predict nurses' intention to administer opioids to these patients. Perceived control was the strongest predictor. Nurses reported positive overall attitudes towards opioids and their use in pain management. However, many negative attitudes were identified; for example, administering the least amount of opioid and encouraging patients to have non-opioids rather than opioids for pain relief. The findings related to specific attitudes and normative pressures provide insight into registered nurses' management of pain for hospitalized patients and the direction for educational interventions to improve registered nurses' administration of opioids for pain management. PMID- 11882192 TI - Literature review of role stress/strain on nurses: an international perspective. AB - The presence of role stress/strain among nurses has been of concern throughout the world. However, to date, no one has conducted, from an international perspective, a literature review of research on the topic. This article assesses research from 17 countries, identifies the major areas of focus in the studies, compares and contrasts the findings, summarizes the state of the science on role stress/strain on nurses and makes recommendations for future research. PMID- 11882193 TI - The criteria of appropriateness and feasibility. PMID- 11882194 TI - Explorations of a trust approach for nursing ethics. AB - Trust has long been acknowledged as central to nurse-patient relationships. It, however, has not been fully explored normatively. That is, trust must be examined from a perspective that encompasses not only reliability and competence, but also good will within nursing relationships. In this paper, we explore how a trust approach, based on Annette Baier's work on trust in feminist ethics, could help inform future developments in nursing ethics. We discuss the limitations of other approaches such as those based on contracts, paternalism, and care. By drawing out central features of Baier's theory, we demonstrate how it can help overcome the problems of these previous models. In doing so, we emphasise the importance of combining the ethics of care and justice, acknowledging vulnerability and the potential for evil in nursing relationships, and politically situating the ethical concerns of nursing. PMID- 11882195 TI - Theory and language: locating agency between free will and discursive marionettes. AB - This article outlines a research methodology that embraces individual narratives, yet recognizes that individual narratives are nested within a backdrop of broader social and cultural understandings of who we are and how we come to understand our world. This dialectical move requires an epistemological shift, focusing on the utility of reconceptualizing the 'environment', not only as the social, political, or economic conditions in society, but also as language. Reconceptualizing the environment as language makes it epistemologically possible to construct a bridge between varying levels of analysis, namely, between individual accounts and life stories and the cultural, social, and historical worlds from which those accounts emerge. PMID- 11882197 TI - Schizophrenia and the 'disquieting' consequences of social poetics: a response to Aldridge and Stevenson. PMID- 11882196 TI - Social poetics as research and practice: living in and learning from the process of research. AB - This paper is both a report of research work carried out by one author of the paper with the other involved in a supervisory role, and a reflection on methodology that was an emergent property of the research process. The research question arose when professional preunderstandings about schizophrenia as a biological disturbance were bracketed as a Husserlian form of phenomenology was adopted. The initial study focused on the meanings three individuals attached to being diagnosed with a mental illness and being involved in psychiatric treatment systems. The exploration of meanings led to the researcher being guided by one study participant in relation to a suitable framework for making sense of her lived experience of being entangled in psychiatric systems. This involved an appreciation of the poetics of interpersonal interaction that was simultaneously fuelled by the researcher's reading of social poetics in psychology. Thus, the research became a hermeneutic inquiry, focused upon the text of one participant (Beth)1 and the psychiatric system that she and colleagues encountered. Beth's psychiatric predicament was described by a loss of 'self', psychiatric interrogation, and positive and negative features of immersion in the system. A byproduct of taking a poetic approach was that the participant found the research process therapeutic. Thus, social poetics encouraged a beneficial blurring of the boundaries between research and practice. In both arenas, social poetics can lead to a different level of understanding. This is contradictory to mainstream approaches to research that value objectivity with a strict division between the researcher and the subjects, with the aim of producing scientific knowledge. PMID- 11882199 TI - The externality of the inside: body images of pregnancy. AB - This paper draws on literature, empirical data and a range of theoretical perspectives on the maternal body to examine understandings of the relationship between a pregnant woman and her foetus, with a particular focus on the body images used by women to represent this relationship. Psychoanalytic and nursing accounts of the relationship between mother and foetus have often described a symbiotic 'oneness' or unity during pregnancy. Such accounts, however, stress the temporary nature of this unity and identify a series of 'stages' of separation or 'polarisation' between mother and foetus during pregnancy. In contrast, many of the 25 women who participated in our interview study of new motherhood described a confusion of the boundary between self and foetus. For many women the experience of pregnancy and the relationship with the unborn baby was ambiguous and uncertain. Importantly, none of these women described her relationship with the foetus as a series of developmental stages, but rather saw it as fluctuating throughout pregnancy. These findings are more consistent with the work of feminist theorists who describe pregnancy as a dynamic and fluid merging of the inside and the outside of the body/self. PMID- 11882200 TI - Environmental influences on the experiences of people with Parkinson's disease. AB - This study elucidates environmental influences on lived illness experiences. For two consecutive years, persons with Parkinson's disease (PD) participated in 1 week of daily walking in the Swedish mountains. Daily, low-intensive walking that is free of intense effort or time pressures associated with group interaction characterized the week. Participants were interviewed 3 months after the mountain stay regarding experiences in the mountains, daily living, and how their experience in the mountains influenced their daily living after returning home. A phenomenological method was used for data analysis. Results point to the close connection between mind, body, and environment. The connection becomes highlighted when people are afflicted with sickness, such as PD, which causes impaired control of body language, impaired voluntary mobility, and lowered energy levels. The results also show how a social context in an environment with suitable physical challenges led to a change in individuals' perceptions of the manageability of their experienced sicknesses. These results provide a deepened understanding of how individuals with PD experience illness, its influences on daily life, and how a suitable environment opens opportunities for managing daily issues. PMID- 11882201 TI - A 'good enough' nurse: supporting patients in a fertility unit. AB - In this paper, I discuss the findings of an ethnographic study of a fertility unit. I suggest that caring as 'emotional awareness' and 'non-caring' as 'emotional distance' may be forms of nursing akin to Fabricius's (1991) arguments around the 'good enough' nurse. This paper critiques caring theories and contributes to the debates over the nature of caring in nursing. I discuss the implications raised for nurses if patients want a practical approach to caring and do not expect an emotionally intimate relationship from nurses. PMID- 11882202 TI - The voices of days gone by: advocating the use of oral history in nursing. PMID- 11882203 TI - The International Academic Nursing On-Line Alliance. PMID- 11882204 TI - The implications of healthcare reforms for the profession of nursing. AB - This paper offers a wide-ranging analysis of concerns that the value of emotion work within nursing is being eroded. We examine the occupation's historical development to argue that, in so far as emotion work has any essence within nursing, it is as an occupational myth which has been deployed to legitimate nurses' jurisdictional claims. We argue that recent developments in health-care raise questions about the benefits of claims of this kind and suggest that a little more realism about the nature of nursing work might make for a more sustainable professional future. PMID- 11882205 TI - Nursing leadership and health sector reform. AB - The political, technological and economic changes that have occurred over the past decade are increasingly difficult to manage within the traditional framework of health-care, and the organisation of health-care is seen to need radical reform to sweep away many of the internal barriers that now divide one form of health-care, and one profession, from another. Nursing must equip itself with skills in advocacy and political action to influence the direction the system will take. Nursing currently suffers from a weakness in self-concept that goes hand in hand with a weakness in political status, and nursing leadership must build the foundations for both advocacy for others and self-advocacy for the nursing movement. The profession faces tensions between different conceptions of its role and status, its relationship to medicine, and its relationship to health. Health indices are tightly linked to status, and to trust, hope, and control of one's own life. Can nurses help empower others when they are not particularly good at empowering themselves? What will the role of the nurse be in creating the information flows that will guide people toward health? Nursing's long history of adaptation to an unsettled and negotiated status may mean that it is better fitted to make this adaptation than other more confident disciplines. PMID- 11882206 TI - Privileged position: preparing nurses to work in the community. AB - Nurses nurse, end of story, but is it? Is nursing any one thing? Nursing is many things and the positions that nurses occupy in any particular setting are multiple. In an educational project aimed at facilitating student learning, I set out to expose the multipositionality of community nursing and to acknowledge that discursive pressures shape and limit practice. A set of resources using story method was produced for use in educational settings. Story method was chosen as an emancipatory vehicle for learning. A worrisome question that arose from reflection on the project was that although the educational aim of the project was emancipatory, did the singularity of the professional voice offered through the stories sabotage that goal? PMID- 11882207 TI - Embodied largeness: a significant women's health issue. AB - This paper describes a three-year long research project in which nine large bodied women have engaged in a prolonged dialogue with the researcher about the experience of being 'obese'. The study involved an extensive review of the multidisciplinary literature that informs our understandings of body size. The literature review was shared with participants in order to support their critical understanding of their experience. An examination of a wide range of literature pertinent to the area of study reveals widespread acceptance of the notion that to be thin is to be healthy and virtuous, and to be fat is to be unhealthy and morally deficient. The experience of participants raised questions as to how nursing could best provide health-care for large women. According to the literature review, nurses have perpetuated an unhelpful and reductionist approach to their care of large women, in direct contradiction to nursing's supposed allegiance to a holistic approach to health-care. This paper suggests strategies for an improved response to women who are concerned about their large body size. PMID- 11882208 TI - The conversational and discursive construction of community psychiatric nursing for chronically confused people and their families. AB - The paper examines the conversational and discursive processes that occur within domiciliary visits between community psychiatric nurses (CPNs) and relatives of chronically confused people. Three conversation formats are identified, through which talk between CPNs and carers is organised. In addition, various discursive practices are also identified within domiciliary meetings. The contribution of these conversational and discursive features to the work of CPNs is fully discussed in relation to what they accomplish within the visit. The data comprised 48 paired tape recordings. Of these recordings, 24 were unstructured interviews between CPNs and informal carers to people who had been diagnosed as having dementia by a medical practitioner. In addition, 24 domiciliary meetings between primary informal carers and their CPNs were tape recorded and fully transcribed. The transcripts were analysed using techniques drawn from conversation analysis and discourse analysis. The implications of the findings upon CPN practice are examined. PMID- 11882209 TI - Nurses and medication error: a discursive reading of the literature. AB - Medication management is a part of everyday nursing practice. In carrying out this aspect of their role, nurses are subject to a range of practices and procedures, which are dictated through legal, management and medical requirements to ensure safe administration of medications. The literature addresses a range of areas related to the nurse's role in medication management, including knowledge required by nurses, practices and procedures related to administration, procedures for identifying and reducing errors and measuring error rates, and professional and ethical issues. The literature is a part of the discursive ensemble of nursing - as a written text it contributes to how nurses and colleagues construct and view nursing practice. This paper presents a discursive reading of that literature, exploring how nurses are positioned in relation to medication errors. I argue that the voice of nursing is mostly heard through discourses of biomedical science, law and management and draw on the work of Michel Foucault to illustrate some of the ways in which these discourses shape nursing practice. PMID- 11882210 TI - The influence of liberal political ideology on nursing science. AB - Previous notions of science as impartial and value-neutral have been refuted by contemporary views of science as influenced by social, political and ideological values. By locating nursing science in the dominant political ideology of liberalism, the author examines how nursing knowledge is influenced by liberal philosophical assumptions. The central tenets of liberal political philosophy - individualism, egalitarianism, freedom, tolerance, neutrality, and a free-market economy - are primarily manifested in relation to: (i) the individualistic focus of our science; (ii) our view of society as essentially egalitarian and equitable; (iii) our preference for politically neutral knowledge development, and (iv) an economy of knowledge development that supports rather than challenges the status quo. I argue that exposing, rather than ignoring, the liberal ideological values inherent in nursing science will render these assumptions open to debate, stimulate ongoing development of critically oriented knowledge, and increase our capacity to influence the social, political and economic determinants of health. PMID- 11882211 TI - Emotional labor and nursing: an under-appreciated aspect of caring work. AB - This paper describes one component of the findings of a larger research study entitled 'Nurses' social construction of self: Implications for work with abused women'. One of the most consistent themes arising from that study involved nurses' views regarding the relevance of emotional engagement/detachment in pursuit of excellence in their practice. In this article this theme is examined in the light of work on emotional labor and the emotional work of nursing. Nurses' high degree of satisfaction in the emotional rewards of their work with clients is described and contrasted with their dissatisfaction in relation to nursing education and their views of the lack of valuing of nurses' work by others within the healthcare system. The importance of supporting them in relation to the emotional aspects of their work is explored. PMID- 11882212 TI - The government of the sick. PMID- 11882213 TI - Managing home nursing care: visibility, accountability and exclusion. AB - The paper examines managerial practices shaping contemporary home nursing care. Foucault's writings on governmentality are used to appraise managerial and nursing practices understood as exemplars of forms of government of people's health. An ethnographic study of organizational practices shaping contemporary home nursing care reveals that the everyday work of managers involves making particular forms of nursing practice visible. Through careful scripting of these visible forms of practice, managers and nurses together work to exclude the local knowledge of patients and of nurses regarding experiences of living with chronic illness. Recommendations are offered for managers and nurses who seek to develop more autonomous roles for nurses: roles that require the inclusion of people's own knowledge of how they live at home with their chronic illness. PMID- 11882214 TI - Governmentality, the iconography of sexual disease and 'duties' of the STI clinic. AB - Sexually transmitted infections (STIs) have come to occupy a different social space over the last hundred years, where the iconography of disease has moved from purity to consumption of desire, and where the regulation of disease has moved from moral proscription to governmentality. These processes are represented through health promotion campaigns where the medico-moral discourses appropriated both the iconography of pathology and the construction of the family as the site of moral surveillance and governmentality. This paper will consider how the 'duties' of STD clinics have been defined and mark a paradigmatic statement of the panoptic role of medicine in the mapping of the social and psychological spaces between individuals. Sexual health medicine, together with health promotion ideologies, has claimed privileged status through the deployment of expert, clinical knowledge and rationalities, to penetrate the sexual praxis of populations. In so doing, it also underscored the individual's roles and responsibilities in the ideological work of the changing constructions of sexual citizenship, from moral purity to ars erotica, as well as the increasing intervention of the state in reproduction and control of sexualities. PMID- 11882215 TI - Discourses of the body in euthanasia: symptomatic, dependent, shameful and temporal. AB - This theoretical paper is derived from a discourse analysis of the textual material from a study of the seven deaths associated with legalised euthanasia in the Northern Territory, Australia. The textual analysis utilises evidence from interviews, letters written by people seeking euthanasia, medical reports, coroner's records and media reports concerning the social experiment of legalised euthanasia in Australia. The paper does not discuss the euthanasia debate. It argues that the body is a neglected concern in the debates and offers a construction of the discourses of the body as symptomatic, dependent, shameful and temporal. Medical discourses frame the body as symptomatic but these people were also concerned with the loss of autonomy associated with dependence, with shame connected with loss of bodily functions and the embodied experience of determining a 'time to die'. PMID- 11882216 TI - A critical analysis of compliance. AB - In nursing the word compliance has competing meanings. In order to understand these meanings, nursing literature was reviewed and a critical analysis of this concept was undertaken. This included an examination of how nursing was located in relation to the historical controversy surrounding the term compliance. The philosophy that undergirds this analysis is critical theory scholarship, which focuses on language as a vehicle for social control and domination. Literature was critically analyzed according to how nurse authors define the term compliance and the historical context in which the term was used. Analysis of the literature revealed three distinct categories: evaluative, rationalization, and acceptance. Each of these categories is described and the selection criteria identified. We recommend that, nurses intent on conducting future compliance research, consider emancipatory models for their investigations. PMID- 11882217 TI - From delegation to specialization: nurses and clinical trial co-ordination. AB - This paper considers an area of clinical research that has been delegated by physician-researchers to nurses and others in the United States, that of clinical trials co-ordination. It uses interviews with nurse trial co-ordinators to explore the occupational processes by which the boundaries of work enactment and the definition of work have been established by nurses and others. It then discusses the occupational processes that have been established to formalize a role for nurses in clinical research. It raises the question of (and offers speculation on) whether specialization alone will distinguish nursing from other occupational groups engaged in clinical research work. PMID- 11882218 TI - Michael Crotty and nursing phenomenology: criticism or critique? AB - In 1996 Michael Crotty published the text Phenomenology and nursing research in which he criticised many nurse researchers' interpretation of the methodology of phenomenology and their utilisation of phenomenology as a method for undertaking qualitative nursing research. Crotty's thesis proposes that the research conducted by nurses is not phenomenology according to the European tradition, but a North American hybrid. Subsequently, debate has occurred amongst nurses as to whether Crotty's work is a scholarly, reasoned critique or a severe, judgmental, fault-finding criticism of nursing research. Considering the increasing utilisation of phenomenology as a methodology that informs nursing research, this debate is an important one and has implications for the conduct of research. This article examines this debate and the implications of Crotty's work for phenomenological research in nursing. PMID- 11882219 TI - Implications of organizational learning for nursing managers from the cultural, interpersonal and systems thinking perspectives. AB - This paper discusses the implications of organizational learning to healthcare administrators, in particular nursing managers, from the cultural, interpersonal and systems thinking perspectives. In the various perspectives the reasons for organizational ineffectiveness and remedies are presented. There is little doubt that pressure for nurses to perform is escalating as healthcare institutions attempt to improve the quality of service through restructuring and change, which leads to greater job dissatisfaction, higher turnover, lower morale and increased industrial actions of nurses. An integrated approach to organizational learning is arguably important for the effective management of nurses during periods of transition. PMID- 11882220 TI - Preparing parents for their child's surgery: preoperative parental information and education. PMID- 11882221 TI - Cardiac trauma in children. PMID- 11882222 TI - Is haemostasis biological screening always useful before performing a neuraxial blockade in children? AB - BACKGROUND: Because of the lack of controlled studies, there is no consensus of opinion about the practice of routine haemostasis tests before neuraxial blockade in children. The purpose of this study was to compare the influence of two different strategies of coagulation evaluation on the incidence of diagnosed coagulopathies leading to a modification of the preoperative or anaesthetic management in children who were scheduled for caudal, epidural or intrathecal block. METHODS: For a 24-month period (period 1, retrospective study, n=751), haemostasis screening was undertaken only after family and personal history and physical examination in all patients. For the following 24 months (period 2, prospective study, n=958), a standardized questionnaire was used. In addition, routine tests (prothrombin, partial thromboplastin time, platelet count) were performed in children who where not yet walking. In older children, coagulation tests were undertaken as in period 1. RESULTS: Overall, 26 significant abnormalities were diagnosed. Coagulation tests were performed in 16.2% (period 1) and 78.2% (period 2) of the children, who were not yet walking. Routine tests did not improve the diagnosis of haemostasis abnormalities justifying a modification of the preoperative and anaesthetic management (2.2% from 406 children in period 1 vs 4.1% from 266 children in period 2). The predictive positive value of routine tests (period 2) was 19%, vs 45% for specific tests (period 1) (P < 0.001). In older children, the use of a standardized form increased the number of haemostasis screenings without improvement of diagnosis leading to modified preoperative management (0.3% from 315 children in period 1 vs 0.5% from 628 children in period 2). CONCLUSIONS: When routine testing is performed in nonwalking children, the screening number increases without leading to a higher number of anaesthetic management changes, suggesting that routine testing does not seem to provide much extra information in the absence of a positive history. PMID- 11882223 TI - The introduction of a paediatric anaesthesia information leaflet: an audit of its impact on parental anxiety and satisfaction. AB - BACKGROUND: A paediatric anaesthesia information leaflet was produced to address preoperative parental anxiety and to facilitate informed parental consent. METHODS: An audit was undertaken to assess the impact of introducing the leaflet. This addressed the information needs and expectations of parents of children undergoing anaesthesia, parental satisfaction with information provision and parental preoperative anxiety. RESULTS: The audit revealed that parents expect to be provided with information, although not necessarily in written form. However, the majority who received the information leaflet concluded that verbal information alone would not have been sufficient. The information leaflet was found to be accessible, informative and useful and those who received it reported greater satisfaction with information provision than a control group. Many parents perceived that it resulted in lower levels of preoperative anxiety CONCLUSIONS: A decision was therefore undertaken that routine use of the leaflet would continue on all of the paediatric surgical wards. However, the study also indicated that leaflets should not replace verbal communication with nursing and medical staff, who remain important sources of information. PMID- 11882224 TI - The effects of an educational programme on the anxiety and satisfaction level of parents having parent present induction and visitation in a postanaesthesia care unit. AB - BACKGROUND: In Hong Kong, some hospitals have established the practice of Parental Presence Induction (PPI) and visitation in Postanaesthesia Care Units (PACU) for children receiving surgery. The literature indicates that parents reported extreme anxiety and discomfort after being present at induction of anaesthesia and suggests that it would seem appropriate to devise a programme of education to reduce anxiety for parents. METHODS: A quasi-experimental pretest and post-test design was employed. Parents in the experimental group received an educational programme about the role and expectations of parents having PPI and visitation in PACU together with an information pamphlet. The comparison group received routine (verbal) instructions. The Chinese version of the State and Trait Anxiety Inventory and the Parental Satisfaction with Care Questionnaire were used to assess parents' anxiety and satisfaction with care. RESULTS: A total of 50 parents (mostly mothers) were recruited. Twenty-five were allocated in the intervention group and 25 in the comparison group. The children were aged 1--9 years with a mean age of 2.9 years. There were 46 male and four female children. The results demonstrate that parents who received an education programme reported a decrease in anxiety (P < 0.001) and an increase in their satisfaction with the care provided (P < 0.001). A significant negative relationship (r= -0.61, P < 0.001) between parental anxiety postoperatively and the satisfaction with care score was also obtained, suggesting that lower levels of parental anxiety are associated with higher levels of satisfaction. CONCLUSIONS: The study suggests that an educational programme preparing parents for their child's induction of anaesthesia and visitation to the PACU should be offered, as it can reduce their anxiety with the practice of PPI and visitation in the PACU and increase parents' satisfaction with care. PMID- 11882225 TI - Comparison of rocuronium and suxamethonium for rapid tracheal intubation in children. AB - BACKGROUND: The purpose of our study was to determine whether a smaller dose of rocuronium than previously reported could provide similar intubating conditions to suxamethonium during rapid-sequence induction of anaesthesia in children. METHODS: One hundred and twenty ASA I, unpremedicated children, aged 1-10 years, who were undergoing elective surgery, were randomized into three groups to receive rocuronium 0.6 mg.kg-1, rocuronium 0.9 mg.kg-1 or suxamethonium 1.5 mg.kg 1. The study was double-blinded, anaesthesia and timing of injection was standardized to alfentanil 10 microg.kg-1, thiopentone 5 mg.kg-1 and the study drug. Intubation was attempted at 30 s after injection of neuromuscular relaxant and intubating conditions graded as excellent, good, poor or impossible. RESULTS: All 120 children were successfully intubated within 60 s without need for a second attempt after administration of neuromuscular relaxant. Differences between suxamethonium and rocuronium 0.6 mg.kg-1 and between the two doses of rocuronium were statistically significant (P=0.016 and 0.007, respectively). CONCLUSIONS: Rocuronium 0.9 mg.kg-1 provides similar intubating conditions to suxamethonium 1.5 mg.kg-1 during modified rapid-sequence induction using alfentanil and thiopentone in children (P=0.671). Rocuronium 0.6 mg.kg-1 was inadequate. PMID- 11882226 TI - A comparison of prilocaine and lidocaine for intravenous regional anaesthesia for forearm fracture reduction in children. AB - BACKGROUND: In this prospective blinded randomized study, we compared prilocaine and lidocaine for intravenous regional anaesthesia for forearm fracture reduction in children. METHODS: Two hundred and seventy-nine children, aged 316 years, were enrolled and randomly assigned to receive 3 mg.kg-1 of either prilocaine or lidocaine. The severity of fracture was classified according to the displacement of the radius (i.e., no radial fracture, angulated, partly displaced or completely displaced). Pain during the procedure was assessed as none, minimal, moderate or severe. RESULTS: There was no significant difference between agents in the proportion of patients with a successful reduction (prilocaine 94%, lidocaine 92%). Compared with less severe fractures, successful reduction was less common in the completely displaced fractures (P < 0.001) but there was no significant difference in this category between anaesthetic agents (successful reduction: prilocaine, 84%; lidocaine, 78%). Analgesia was superior in the lidocaine group with more patients having no or minimal pain (prilocaine, 78%; lidocaine, 90%, P < 0.05). CONCLUSIONS: Both agents are effective for forearm fracture reduction in children with a high incidence of successful reductions, particularly in the minimally or nondisplaced fractures. Lidocaine provided superior analgesia. PMID- 11882227 TI - Effects of rocuronium pretreatment on muscle enzyme levels following suxamethonium. AB - BACKGROUND: The effects of different time intervals between rocuronium pretreatment and suxamethonium administration on muscle-derived enzymes and myoglobin were evaluated. METHODS: Fifty-two patients, aged 3-6 years, were allocated randomly to receive pretreatment of either rocuronium 1 min or 4 min, or a placebo, before suxamethonium. Serum creatine-phosphokinase (CK), lactate dehydrogenase (LDH), alanine aminotransferase, aspartate aminotransferase (AST) and myoglobin concentrations were measured before treatment and at 30 min, 6 h and 24 h after suxamethonium administration. RESULTS: Mean serum CK and myoglobin concentrations in the rocuronium groups were significantly less than in the saline group 24 h and 30 min after suxamethonium administration, respectively (P < 0.05). There were no significant differences between the rocuronium groups. Mean LDH and AST concentrations were higher at 6 h and 24 h after suxamethonium administration compared with preadministration levels, respectively (P < 0.05). CONCLUSIONS: We concluded that intervals of 1 min or 4 min between the rocuronium and suxamethonium administrations have the same effect on enzyme levels. PMID- 11882228 TI - Interleukin-8 secretion following cardiopulmonary bypass in children as a marker of early postoperative morbidity. AB - BACKGROUND: Interleukin (IL)-8, an 8 kDa peptide, is the first chemoattractant identified as being specific for neutrophils. Its possible association with early postoperative morbidity following cardiopulmonary bypass (CPB) in infants and children is unknown. This prospective cohort study sought possible roles of IL-8 in the inflammatory response to CPB and investigated if changes in IL-8 levels and clinical course and outcome were related. METHODS: IL-8 levels were measured in 16 children undergoing CPB. Systemic blood was collected after induction of anaesthesia (baseline), at 15 min after CPB onset and cessation, and at 1, 4, 8, 12 and 24 h thereafter. RESULTS: Correlation coefficients between IL-8 levels and CPB time ranged from 0.45 to 0.55, heart rate 0.41--0.44, surgical time 0.41- 0.63 and pH --0.56 to --0.50 (P < 0.05 for all parameters). Univariate analyses showed that patients requiring inotropic support and those with tachycardia had significantly higher postoperative IL-8 levels (P < 0.05). Furthermore, IL-8 levels were significantly higher in patients with surgical times > 200 min and in patients with an aortic clamp in place for > 65 min. CONCLUSIONS: There was an association between IL-8 and early postoperative heart rate, and the need for inotropic support IL-8 correlated positively with surgical time, CPB time and heart rate and negatively with pH. IL-8 release may be related to some of the haemodynamic changes in the early postoperative course following CPB. The relationship between IL-8 and late markers of patient outcome in high-risk infants awaits further studies. PMID- 11882229 TI - Comparison of pre- and postoperative administration of ketoprofen for analgesia after tonsillectomy in children. AB - BACKGROUND: Tonsillectomy is commonly performed in children, but unfortunately it is associated with intense postoperative pain. The use and optimal timing of nonsteroidal anti-inflammatory drugs (e.g. ketoprofen) during tonsillectomy is controversial. METHODS: We evaluated the safety and efficacy of ketoprofen in 109 children, aged 3-16 years, during and after tonsillectomy in 1998-2000. Standardized anaesthesia was used. Forty-seven children received ketoprofen 0.5 mg.kg-1 at induction (preketoprofen group) and 42 children after surgery (postketoprofen group), followed by continuous ketoprofen infusion of 3 mg.kg-1 over 24 h in both groups; 20 children received normal saline (placebo group). Oxycodone was used for rescue analgesia. RESULTS: Pre- and postketoprofen groups did not differ in experienced pain or in opioid consumption in the first 24 h after surgery; demonstrating that ketoprofen did not have a pre-emptive effect. Patients in the placebo group received 30 more oxycodone doses than did patients in the ketoprofen groups, but the difference was not significant (P=0.074). Two patients (5) in the postketoprofen group had postoperative bleeding at 4 h and 26 h, respectively. Both patients required electrocautery to stop bleeding. Neither the incidence nor the severity of adverse events differed between study groups. CONCLUSIONS: This study demonstrates that ketoprofen did not have a preemptive effect and, at the dose used, did not perform statistically significantly better than placebo. PMID- 11882230 TI - Vincristine-induced vocal cord paralysis in an infant. AB - We report the development of stridor and dysphagia in a 5-month-old-infant with acute lymphoblastic leukaemia after the administration of four weekly doses of vincristine during induction therapy. Because direct laryngoscopy revealed bilateral vocal cord paralysis, the patient underwent elective intubation. Extubation was performed 7 days later, after direct laryngoscopy confirmed recovery of vocal cord mobility. Vincristine-induced bilateral recurrent laryngeal nerve paralysis is a rare but potentially life-threatening complication. Therefore, it should be suspected when stridor is present, and clinicians should consider visualization of the airway to establish the cause of upper airway compromise in infants receiving vincristine. PMID- 11882231 TI - Initial experience with dexmedetomidine in paediatric-aged patients. AB - We present preliminary clinical experience with the use of dexmedetomidine, an alpha2 adrenergic agonist, in children. Dexmedetomidine was administered in three clinical scenarios: (i) to provide sedation during mechanical ventilation; (ii) intraoperatively for controlled hypotension during an orthopaedic surgical procedure; and (iii) to provide sedation during an invasive procedure. Preliminary data from studies in the adult population, its physiological effects and potential applications in paediatric anaesthesia and critical care are discussed. PMID- 11882232 TI - Anaesthesia for phaeochromocytoma removal in a 5-year-old boy. AB - We describe the case of a 5-year-old boy with phaeochromocytoma of the left adrenal gland, treated surgically by removal of the tumour under general anaesthesia. Phaeochromocytoma is a particularly rare tumour in children and surgical excision is the definitive treatment. We discuss the clinical and laboratory characteristics of the case, the diagnostic approach, the preoperative and intraoperative management and the postoperative course. PMID- 11882233 TI - Complicated negative pressure pulmonary oedema in a child with cerebral palsy. AB - A 3-year-old child with cerebral palsy developed postextubation upper airway obstruction secondary to laryngospasm and/or masseteric spasm,which may have been triggered by the muscular spasticity and the slow recovery from inhalational anaesthesia associated with cerebral palsy. This upper airway obstruction was followed by negative pressure pulmonary oedema. The patient improved on mechanical ventilation; however, his condition was complicated with the occurrence of bilateral pneumothoraces. After release of the pneumothoraces and reexpansion of the lungs, the child developed reexpansion pulmonary oedema, culminating in acute lung injury. PMID- 11882234 TI - Perioperative management of a child with very-long-chain acyl-coenzyme A dehydrogenase deficiency. AB - Very-long-chain acyl-coenzyme A dehydrogenase deficiency is an inborn error of fatty acid metabolism. The clinical presentation of this disease in children is either a severe form with onset of symptoms in the first months of life, cardiomyopathy, metabolic acidosis, myopathy and a high mortality, or a less severe form manifesting mainly with hypoglycaemia. Perioperative fasting and (even emotional) stress can trigger metabolic decompensation through the altered metabolism of endogenous fatty acids resulting in hypoglycaemia, acute cardiac and hepatic dysfunction and rhabdomyolysis. We report the perioperative management of a 9-year-old boy suffering from the severe form of this disease who underwent circumcision. Metabolism was kept stable in this child by using a glucose--electrolyte infusion throughout the perioperative period to avoid the biochemical consequences of fasting and a benzodiazepine--opioid technique combined with regional anaesthesia to minimize the stress response. Considering reports about a possible interference of propofol with fatty acid oxidation and to avoid the unnecessary administration of fatty acids, propofol should not be used in these patients. PMID- 11882235 TI - Caudal injections for postoperative analgesia in children. PMID- 11882237 TI - Cracking loss of continuous positive airway pressure. PMID- 11882238 TI - Epidural analgesia for patients undergoing hepatic portoenterostomy (Kasai procedure). PMID- 11882239 TI - Comparison of external and internal pelvic measurements of Belgian Blue cattle from sample herds in Belgium and the United kingdom. AB - Measurements describing pelvic conformation of pedigree Belgian Blue cows were obtained from a sample of nine herds in Flanders, Belgium, comprising 111 adult breeding cows, and from 11 herds in the United Kingdom comprising 108 similar cows. All herds in the Belgian sample (111 cows) managed parturition by elective caesarian section as did seven herds (56 cows) in the UK sample; the remainder of UK herds (52 cows) allowed cows the opportunity to calve naturally per vagina before resorting to caesarian section. The data described the external and internal measurements, and pelvic area characteristic of this breed, irrespective of the different selection pressures applied over a number of years to this breed through variation in farm management and market forces present in either country. From these data, generalized linear models were constructed to predict pelvic area; they correctly identified cows with either small or large pelvic areas. There were no significant differences in pelvic conformation between cattle bred either in Belgium or the UK, although those cows bred in herds where natural calving was allowed to take place had significantly larger internal pelvic height (p < 0.05) and area (p < 0.05) than other Belgian Blue cows. The correlation coefficients between internal pelvic height and width and external pelvic measurements were significant (p < 0.001). These results might facilitate the selection of breeding cows with larger pelvic area so that a higher proportion of cows can calve naturally than currently occurs. However, selection for other traits such as relatively low birth weight combined with higher weaning weight should be carried out at the same time. PMID- 11882240 TI - Modulation of ovarian function in female dogs immunized with bovine luteinizing hormone receptor. AB - Adult female dogs were immunized with 0.5 mg bovine luteinizing hormone receptor (LH-R) encapsulated in a silastic subdermal implant and subsequently with four intramuscular booster injections of 0.1 mg LH-R each. Circulating LH-R antibody was detected in the sera 3 weeks post-implant. The appearance of LH-R antibody was associated with a decline in the serum progesterone concentrations to a range of 0-0.5 ng/ml until day 365 in the immunized dogs in comparison with a range of 5-10 ng in the control animals, suggesting a lack of ovulation and corpus luteum function in immunized dogs. The immunized dogs did not show signs of 'standing heat' and failed to ovulate when induced by LH-RH challenge. Serum oestradiol levels, however, remained in the range of 30-40 pg/ml in both the immunized and the control dogs. With the decline in the antibody titres, the hormonal profile and vaginal cytology returned to a fertile state and the dogs exhibited signs of 'standing heat', as well as vaginal bleeding. Dogs immunized with LH-R did not show any serious metabolic, local or systemic adverse effects. The hypothalamic- pituitary gonadal axis remained intact as indicated by little difference in pituitary LH levels between control and immunized animals, and by the release of LH by LH-RH challenge. These studies demonstrate that active immunization of female dogs with LH-R could immunomodulate ovarian function to cause a reversible state of infertility. It may be postulated that, due to extensive interspecies homology, a recombinant LH receptor-based immunocontraceptive vaccine may also be effective in other vertebrates. PMID- 11882241 TI - Maturation status, protein synthesis and developmental competence of oocytes derived from lambs and ewes. AB - The efficacy of oocyte selection for in vitro embryo production depends on the abundance and diameter of follicles, cumulus layers around the oocytes and subsequent fertilization. Application of 'ovum pick-up' technique allows us to utilize partially matured oocytes for embryo production even from juvenile subjects. To compare their developmental competence, oocytes derived from lambs and ewes and cultured in maturation medium for up to 26 h were assessed at 2 h intervals by confocal microscopy after chromatin and microtubulin-specific fluorochrome labelling. Lamb oocytes reached second meiotic metaphase (MII) at lower numbers at 24 h (60.0%) and 26 h (28.6%) whereas 85.7% of adult-derived oocytes attained MII status by 24 h of maturation. Radiolabelling of oocyte proteins revealed higher incorporation of [(35)S-]-methionine and [(35)S] cysteine in adult-derived oocytes compared to lamb oocytes. Although the cleavage rate of lamb oocytes was similar to that of ewe oocytes, the proportion reaching blastocyst stage was significantly lower (p < 0.05) in the lamb-derived oocytes. However, blastocysts from both types of oocytes displayed similar cell lineage allocations to inner cell mass and trophectoderm. PMID- 11882242 TI - Placental release/retention in cows and its relation to peroxidative damage of macromolecules. AB - The disturbances in metabolic pathways reflected in clinical symptoms of illnesses may be connected, among others, with the imbalance between production and neutralization of reactive oxygen species. One of such illnesses may be the retention of fetal membranes in cows. The levels of reactive oxygen species can be measured directly or estimated indirectly by the determination of enzymatic and non-enzymatic antioxidative defence systems. The determination of parameters indicating the intensity of peroxidative processes of lipids, proteins and nucleic acids caused by reactive oxygen species is also useful. This review examined the available literature regarding peroxidative processes of lipids, proteins and nucleic acids caused by reactive oxygen species as well as parameters indicating its intensity. All information relates the importance of proper and improper placental release in cows. PMID- 11882243 TI - Influence of puerperal uterine infection on uterine involution and postpartum ovarian activity in dairy cows. AB - This article presents the results of a clinical trial designed to study the effect of puerperal uterine infection on uterine involution and on ovarian activity in dairy cows, monitored twice weekly from parturition until the sixth week postpartum (wpp). Infection significantly retarded uterine involution assessed by the uterine body diameter and a score of intrauterine fluid volume (IUFV). By the sixth wpp, cows with normal puerperium (controls) and cows that showed mild puerperal endometritis had similar uterine body diameter and IUFV, indicating spontaneous recovery within the postpartum voluntary waiting period. However, in cows with severe puerperal endometritis, although uterine body diameter had regressed to pregravid size, IUFV remained significantly higher than in control and mild endometritis cows, indicating that chronic endometritis was established. The IUFV score was positively and significantly correlated with uterine swab bacterial growth density and allowed diagnosis of endometritis after the third wpp. Cows with mild or severe endometritis had a significantly higher prevalence and persistence of pathogenic bacteria (Escherichia coli, Actinomyces pyogenes, Gram-negative anaerobes - GNA) than controls. Actinomyces pyogenes was associated to GNA in 74% of isolations. Ovarian activity measured by ultrasound scanning of the ovaries and plasma progesterone (P4) concentrations was more abnormal (prolonged anoestrus, prolonged luteal phases and ovarian cysts) in cows with severe endometritis than in controls. PMID- 11882244 TI - Effect of feeding and body condition score on multiple ovulation and embryo production in zebu cows. AB - The aim of this study was to standardize the feeding regimen and the body condition score (BCS) for maximum superovulatory responses in indigenous zebu cows. Ten regularly cycling 5-8-year-old dry cows, weighing 176--260 kg with BCS 2.5--4.5 were divided into two equal groups at random. The groups were maintained on either a good-nutrition or a high-nutrition diet. The feedstuffs were analysed by proximate feed analysis and the metabolizable energy content was estimated. After 3 months feeding, individual cows were injected (i.m) with 1500 IU pregnant mare's serum gonadotrophin (PMSG) at day 10 or day 11 of the oestrous cycle (day of oestrus = day 0). Alfaprostol (6 mg) was injected (i.m) 48 h after the injection of PMSG to induce oestrus. At day 6 or day 7 (day of insemination = day 1), the contents of individual uterine horns were flushed with 150-200 ml of phosphate-buffered saline + 0.2% bovine serum albumin using a two-way Foley catheter. The embryos were identified, evaluated and graded as excellent, good, fair or poor under a stereomicroscope. For the good- and high-nutrition diets, the daily intake of green grass, straw, concentrate, dry matter, crude protein and estimated metabolizable energy by individual cows were 5 and 6 kg, 3 and 3 kg, 1.5 and 3.5 kg, 4.87 and 6.82 kg, 0.39 and 0.74 kg, and 39.60 and 59.12 MJ, respectively. The protein content was 8 and 11% in the good- and high-nutrition diets, respectively. The two groups of cows on different nutritional diets differed significantly with regard to body weight, body condition score and number of palpated corpora lutea (p < 0.01). For cows on the good-nutrition diet, the median number of recovered embryos and transferable quality embryos were three and two, respectively. The recovery rate of embryos was 79.30% of palpated corpora lutea. Cows on the high-nutrition diet did not yield any embryos. The indigenous zebu cows fed on the good-nutrition diet with BCS 2.5-3 were considered suitable for the induction of superovulation, the cows on the high nutrition diet with BCS 4-4.5 were unsatisfactory and were more prone to cyst formation in the ovaries. PMID- 11882245 TI - 15-Ketodihydro-PGF(2 alpha), progesterone and uterine involution in primiparous cows with induced retained placenta and post-partal endometritis treated with oxytetracycline and flunixin. AB - Retention of the foetal membranes (RFM) and post-partal endometritis are common problems in dairy cows. Among other things, the disease is characterized by a bacterial endometritis with aerobic as well as anaerobic bacteria. From an endocrine perspective, cows with RFM have high levels of 15-ketodihydro-PGF(2 alpha) (PG-metabolite) immediately after parturition but these levels fall rapidly within 2 weeks post-partum (early PG-metabolite elevation). After this decline, the PG-metabolite levels increase again and the levels (at this time of a lower magnitude) remain elevated during the period of uterine infection (late PG-metabolite elevation). The aim of this study was to investigate the PG metabolite profiles in cows with retained placenta and post-partal endometritis treated with the prostaglandin synthesis inhibitor flunixin (F), either alone or in combination with oxytetracycline (T). The study was accomplished over 2 years with 12 primiparous cows in each experiment. As a model for RFM, preterm parturition was induced in late-pregnant heifers by injecting PGF(2 alpha) (25 mg i.m) twice with a 24 h interval. In each experiment, the cows were divided into four groups and treated with either T (10 mg/kg b.w. i.m. once daily), F (2.2 mg/kg b.w. p.o. twice per day), a combination of T and F (dosage, as above), or conservatively (0). The treatment periods lasted from day 11 to day 14 post partum (pp) in experiment 1 (after placental shedding, groups T1, F1, TF1 and 0) and from day 3 to day 6 pp in experiment 2 (before placental shedding, groups T2, F2, TF2 and 0). Jugular vein blood samples were collected for analyses of PG metabolite and flunixin. Uterine biopsies were collected twice weekly for investigation of endometrial microbiology. Rectal palpation and ultrasonographic examinations were performed three times per week for investigations of uterine and cervical involution and ovarian activity. No attempts were made to remove the placentas manually. The experiment lasted until day 56 pp. The induction of parturition was successful in all heifers and 22 of 24 animals had RFM. All RFM cows had bacterial endometritis, based on bacteriological examinations. Flunixin treatment (F1, TF1, F2 and TF2) suppressed PG-metabolite levels significantly (p=0.006) during the period of treatment in both experiments. However, the early flunixin treatment only suppressed PG synthesis partially. Late oxytetracycline treatment (T1) did not influence the PG-metabolite levels but oxytetracycline treatment (T2 and TF2) before placental shedding significantly altered the kinetics of the early PG-metabolite elevation compared with other treatments. Late PG-metabolite elevation was significantly correlated to duration of uterine infection and cervical involution. In conclusion, flunixin treatment of cows with retained placenta either before or after placental shedding suppresses prostaglandin synthesis. However, early treatment, when the release of prostaglandins is high, might need more intensive treatment in order to prevent the PG synthesis effectively. Oxytetracycline treatment during the period immediately after parturition before placental shedding might influence the PG metabolite profile and suggests a bacteriological contribution to the high levels of PG-metabolite seen during the first 2 weeks pp in cows with retained placenta. In this study, a correlation between prostaglandin release, the final cervical involution and the end of infection was found. This suggests a link between uterine endocrinology, bacteriology and involution in cows with retained placenta. PMID- 11882246 TI - Reduced phagocytotic activity of macrophages in the bovine retained placenta. AB - This study was carried out to investigate the distribution of immune cells in the bovine placenta during the postpartum period and to compare these cells between normal and retained placenta. Within 1 h after normal calving, biopsy samples of placentomes were collected from 10 cows. The occurrence of retention of fetal membranes was monitored for more than 8 h post-calving, and the samples obtained were divided into two groups: normally discharged and retained placenta (n = 5 each). Immunohistochemical procedures were utilized to detect macrophages and T lymphocytes. Numerous CD14-positive macrophages were found in the stroma of both normal placenta and retained placenta whereas only a few CD3-positive T lymphocytes were found in both cases. However, histochemical staining for acid phosphatase, a predominant lysozomal enzyme, revealed that almost all macrophages showed strong enzyme activity in the normally discharged placentas, whereas in retained placenta the activity of acid phosphatase was conspicuously decreased in intensity. These results indicate that there are functional differences in placental macrophages between normal and retained placenta. PMID- 11882247 TI - Treatment of unobserved oestrus in a dairy cattle herd with low oestrous detection rate up to 60 days post-partum. AB - The efficiency of treatments for unobserved oestrus and their effect on the reproductive performance of a dairy cattle herd with low oestrous detection rate till 60 days post-partum (dpp), attributed to the declivous and slippery concrete floor were investigated. The herdsman requested advice in order to improve the mean days open of the herd, but no investments were allowed because a new unit was about to be built. Due to the low oestrus detection rate of the herd, the breeding policy was to inseminate at the first detected post-partum oestrus. Cows were examined at 20-30 dpp to assess uterine involution, ovarian activity and prevalence of reproductive disorders and, at 60 dpp if no previous oestrus was detected. Each examination included palpation per rectum, ultrasound scanning and collection of a blood sample for plasma progesterone (P4) measurement. Cows with unobserved oestrus till 60 dpp were allocated either to a treatment group (n=139) or to a control group (n=139). Three treatments were used: (a) injection of PGF(2 alpha) (PG) upon detection of a corpus luteum (CL; n = 30), cows not observed in oestrus being re-injected 11-12 days later. AI was at oestrus; (b) PRID (n=35) or Crestar (n=74) devices kept in situ for 12 and 9 days, respectively, were associated to an injection of PG and of equine chorionic gonadotrophin (eCG) at device removal. Cows were double-fixed time-inseminated at 48 and 72 h after device removal. All treated cows were examined at 48-72 h after treatment to confirm oestrus. The percentage of cows detected in oestrus up to 60 dpp remained unchanged through the trial (35 and 47% for years before intervention: 1994-95; 51 and 48% for years of intervention: 1996-97). In contrast, the oestrous detection rate was high both in treated (93%) and control (100%) cows. This possibly resulted from an improvement in the oestrous detection efficiency of the herd's personnel and from examination of cows at 48-72 h after treatment. Treated and control cows had identical conception rate (CR; 36 and 37%, respectively) and reproductive performance. However, the mean days open of the herd in 1996 was significantly improved in comparison with previous years (mean +/- SEM: 134 +/- 6, 126 +/- 5, 110 +/- 4 and 114 +/- 5 days, for years 1994, 1995, 1996 and 1997, respectively, p < 0.05, ANOVA). Conception rate to AI up to 40 dpp was significantly reduced, compared with the period between 60 and 100 dpp but, mean days open were significantly improved in cows inseminated up to 60 dpp, compared with thereafter (p < 0.05). PMID- 11882249 TI - Editorial. PMID- 11882248 TI - Within-herd use of boar semen at 5 degrees C, with a note on electronic monitoring of oestrus. AB - A system was designed to allow a small swine farm in a northern latitude to use its own boars for artificial insemination (AI) conveniently. Semen was collected twice weekly for 3 day use (days 0, 1 and 2), extended in an egg yolk extender and stored at 5 degrees C. Farm personnel were trained to manage the entire AI programme. For simplicity all semen collected was used for insemination. In the first test 47 gilts and 15 sows were inseminated with semen from four boars. One boar was subfertile with a farrowing rate of 36%. The averages for the other boars ranged from 71 to 100%. Then semen was collected from seven boars and all was used to inseminate 70 gilts and 55 sows with 3 x 10(9) or more sperm. Overall 63% farrowed an average of 10.1 piglets per litter. Litter size for sows was 1.5 piglets larger than for gilts. There was no difference in farrowing rate when more than 3 x 10(9) sperm were inseminated. The feasibility of initiating a complete AI programme within a small herd using herd boars was established. However, selection of the boars, use of only high quality semen, and experience with detecting oestrus was required to increase the farrowing rate. The use of various agents to protect sperm against cold shock below 15 degrees C is worthy of further investigation. A new type of electronic probe, which measures the conductivity of cervical mucus, could be helpful if a boar is not available for conventional detection of oestrus. PMID- 11882250 TI - tacg--a grep for DNA. AB - BACKGROUND: Pattern matching is the core of bioinformatics; it is used in database searching, restriction enzyme mapping, and finding open reading frames. It is done repeatedly over increasingly long sequences, thus codes must be efficient and insensitive to sequence length. Such patterns of interest include simple motifs with IUPAC degeneracies, regular expressions, patterns allowing mismatches, and probability matrices. RESULTS: I describe a small application which allows searching for all the above pattern types individually, which further allows these atomic motifs to be assembled into logical rules for more sophisticated analysis. CONCLUSION: tacg is small, portable, faster and more capable than most alternatives, relatively easy to modify, and freely available in source code. PMID- 11882251 TI - An algorithm and program for finding sequence specific oligonucleotide probes for species identification. AB - BACKGROUND: The identification of species or species groups with specific oligo nucleotides as molecular signatures is becoming increasingly popular for bacterial samples. However, it shows also great promise for other small organisms that are taxonomically difficult to tract. RESULTS: We have devised here an algorithm that aims to find the optimal probes for any given set of sequences. The program requires only a crude alignment of these sequences as input and is optimized for performance to deal also with very large datasets. The algorithm is designed such that the position of mismatches in the probes influences the selection and makes provision of single nucleotide outloops. Program implementations are available for Linux and Windows. PMID- 11882252 TI - CFEOM1, the classic familial form of congenital fibrosis of the extraocular muscles, is genetically heterogeneous but does not result from mutations in ARIX. AB - BACKGROUND: To learn about the molecular etiology of strabismus, we are studying the genetic basis of 'congenital fibrosis of the extraocular muscles' (CFEOM). These syndromes are characterized by congenital restrictive ophthalmoplegia affecting muscles in the oculomotor and trochlear nerve distribution. Individuals with the classic form of CFEOM are born with bilateral ptosis and infraducted globes. When all affected members of a family have classic CFEOM, we classify the family as a CFEOM1 pedigree. We have previously determined that a CFEOM1 gene maps to the FEOM1 locus on chromosome 12cen. We now identify additional pedigrees with CFEOM1 to determine if the disorder is genetically heterogeneous and, if so, if any affected members of CFEOM1 pedigrees or sporadic cases of classic CFEOM harbor mutations in ARIX, the CFEOM2 disease gene. RESULTS: Eleven new CFEOM1 pedigrees were identified. All demonstrated autosomal dominant inheritance, and nine were consistent with linkage to FEOM1. Two small CFEOM1 families were not linked to FEOM1, and both were consistent with linkage to FEOM3. We screened two CFEOM1 families consistent with linkage to FEOM2 and 5 sporadic individuals with classic CFEOM and did not detect ARIX mutations. CONCLUSIONS: The phenotype of two small CFEOM1 families does not map to FEOM1, establishing genetic heterogeneity for this disorder. These two families may harbor mutations in the FEOM3 gene, as their phenotype is consistent with linkage to this locus. Thus far, we have not identified ARIX mutations in any affected members of CFEOM1 pedigrees or in any sporadic cases of classic CFEOM. PMID- 11882253 TI - Digital analysis of cDNA abundance; expression profiling by means of restriction fragment fingerprinting. AB - BACKGROUND: Gene expression profiling among different tissues is of paramount interest in various areas of biomedical research. We have developed a novel method (DADA, Digital Analysis of cDNA Abundance), that calculates the relative abundance of genes in cDNA libraries. RESULTS: DADA is based upon multiple restriction fragment length analysis of pools of clones from cDNA libraries and the identification of gene-specific restriction fingerprints in the resulting complex fragment mixtures. A specific cDNA cloning vector had to be constructed that governed missing or incomplete cDNA inserts which would generate misleading fingerprints in standard cloning vectors. Double stranded cDNA was synthesized using an anchored oligo dT primer, uni-directionally inserted into the DADA vector and cDNA libraries were constructed in E. coli. The cDNA fingerprints were generated in a PCR-free procedure that allows for parallel plasmid preparation, labeling, restriction digest and fragment separation of pools of 96 colonies each. This multiplexing significantly enhanced the throughput in comparison to sequence-based methods (e.g. EST approach). The data of the fragment mixtures were integrated into a relational database system and queried with fingerprints experimentally produced by analyzing single colonies. Due to limited predictability of the position of DNA fragments on the polyacrylamid gels of a given size, fingerprints derived solely from cDNA sequences were not accurate enough to be used for the analysis. We applied DADA to the analysis of gene expression profiles in a model for impaired wound healing (treatment of mice with dexamethasone). CONCLUSIONS: The method proved to be capable of identifying pharmacologically relevant target genes that had not been identified by other standard methods routinely used to find differentially expressed genes. Due to the above mentioned limited predictability of the fingerprints, the method was yet tested only with a limited number of experimentally determined fingerprints and was able to detect differences in gene expression of transcripts representing 0.05% of the total mRNA population (e.g. medium abundant gene transcripts). PMID- 11882255 TI - Expression of a crown gall biological control phenotype in an avirulent strain of Agrobacterium vitis by addition of the trifolitoxin production and resistance genes. AB - BACKGROUND: Agrobacterium vitis is a causal agent of crown-gall disease. Trifolitoxin (TFX) is a peptide antibiotic active only against members of a specific group of alpha-proteobacteria that includes Agrobacterium and its close relatives. The ability of TFX production by an avirulent strain of Agrobacterium to reduce crown gall disease is examined here. RESULTS: TFX was shown to be inhibitory in vitro against several A. vitis strains. TFX production, expressed from the stable plasmid pT2TFXK, conferred biological control activity to an avirulent strain of A. vitis. F2/5, against three virulent, TFX-sensitive strains of A. vitis tested on Nicotiana glauca. F2/5(pT2TFXK) is significantly reduces number and size of galls when co-inoculated with tumorigenic strain CG78 at a 10:1 ratio, but is ineffective at 1:1 or 1:10 ratios. F2/5(pT2TFXK) is effective when co-inoculated with tumorigenic strain CG435 at 10:1 and 1:1 ratios, but not at a 1:10 ratio. When F2/5(pT2TFXK) is co-inoculated with CG49 at a 10:1 ratio, the incidence of gall formation does not decline but gall size decreases by more than 70%. A 24 h pre-inoculation with F2/5(pT2TFXK) does not improve biological control at the 1:10 ratio. CONCLUSIONS: TFX production by an avirulent strain of Agrobacterium does confer in that strain the ability to control crown gall disease on Nicotiana glauca. This is the first demonstration that the production of a ribosomally synthesized, post-translationally modified peptide antibiotic can confer reduction in plant disease incidence from a bacterial pathogen. PMID- 11882256 TI - Comparison of the sensitivity of a 24 h-shell vial assay, and conventional tube culture, in the isolation of Herpes simplex virus - 1 from corneal scrapings. AB - BACKGROUND: Herpes simplex keratitis is a sight threatening ocular infection. A rapid and specific diagnosis is essential for the institution of specific antiviral therapy and to avoid complications that can arise from misdiagnosis and inappropriate treatment. Though a variety of techniques are available, isolation of Herpes simplex virus 1 (HSV-1) in culture provides the most reliable and specific method, and is considered as the gold standard in laboratory diagnosis of herpes simplex keratitis. We report a comparative study of the sensitivity of a 24 h-shell vial assay and conventional tube culture in the isolation of HSV-1 from corneal scrapings. METHODS: A total of 74 corneal scrapings obtained from 74 patients with a clinical suspicion of herpes simplex keratitis submitted for the isolation of HSV-1, were simultaneously inoculated into shell vial and tube cultures employing the vero cell line. Shell vial and tube cultures were terminated at 24 h and fifth day respectively. Isolation of HSV-1 was confirmed employing an indirect immunofluorescence assay. RESULTS: HSV-1 was isolated from 24/74 (32.4%) specimens employing both the methods. Sensitivity of both the techniques were found to be similar (20/24, 83.3%) (P = 1.0). CONCLUSION: A 24 h shell vial assay is a rapid alternative technique in comparison to the time consuming conventional tube cultures for the isolation of HSV-1, especially from corneal scrapings for the laboratory diagnosis of herpes simplex keratitis. PMID- 11882254 TI - The Siblings With Ischemic Stroke Study (SWISS) protocol. AB - BACKGROUND: Family history and twins studies suggest an inherited component to ischemic stroke risk. Candidate gene association studies have been performed but have limited capacity to identify novel risk factor genes. The Siblings With Ischemic Stroke Study (SWISS) aims to conduct a genome-wide scan in sibling pairs concordant or discordant for ischemic stroke to identify novel genetic risk factors through linkage analysis. METHODS: Screening at multiple clinical centers identifies patients (probands) with radiographically confirmed ischemic stroke and a family history of at least 1 living full sibling with stroke. After giving informed consent, without violating privacy among other family members, the proband invites siblings concordant and discordant for stroke to participate. Siblings then contact the study coordinating center. The diagnosis of ischemic stroke in potentially concordant siblings is confirmed by systematic centralized review of medical records. The stroke-free status of potentially discordant siblings is confirmed by validated structured telephone interview. Blood samples for DNA analysis are taken from concordant sibling pairs and, if applicable, from 1 discordant sibling. Epstein-Barr virus-transformed lymphoblastoid cell lines are created, and a scan of the human genome is planned. DISCUSSION: Conducting adequately powered genomics studies of stroke in humans is challenging because of the heterogeneity of the stroke phenotype and the difficulty of obtaining DNA samples from clinically well-characterized members of a cohort of stroke pedigrees. The multicentered design of this study is intended to efficiently assemble a cohort of ischemic stroke pedigrees without invoking community consent or using cold-calling of pedigree members. PMID- 11882257 TI - What kind of evidence is it that Evidence-Based Medicine advocates want health care providers and consumers to pay attention to? AB - BACKGROUND: In 1992, Evidence-Based Medicine advocates proclaimed a "new paradigm", in which evidence from health care research is the best basis for decisions for individual patients and health systems. Hailed in New York Times Magazine in 2001 as one of the most influential ideas of the year, this approach was initially and provocatively pitted against the traditional teaching of medicine, in which the key elements of knowing for clinical purposes are understanding of basic pathophysiologic mechanisms of disease coupled with clinical experience. This paper reviews the origins, aspirations, philosophical limitations, and practical challenges of evidence-based medicine. DISCUSSION: EBM has long since evolved beyond its initial (mis)conception, that EBM might replace traditional medicine. EBM is now attempting to augment rather than replace individual clinical experience and understanding of basic disease mechanisms. EBM must continue to evolve, however, to address a number of issues including scientific underpinnings, moral stance and consequences, and practical matters of dissemination and application. For example, accelerating the transfer of research findings into clinical practice is often based on incomplete evidence from selected groups of people, who experience a marginal benefit from an expensive technology, raising issues of the generalizability of the findings, and increasing problems with how many and who can afford the new innovations in care. SUMMARY: Advocates of evidence-based medicine want clinicians and consumers to pay attention to the best findings from health care research that are both valid and ready for clinical application. Much remains to be done to reach this goal. PMID- 11882258 TI - Electronic artificial hand controlled by reconstructed digit. AB - OBJECTIVE: To treat the loss of part of the forearm with a multi-dimension freedom electronic artificial hand, which is controlled by a reconstructed finger transplanted from the second toe to the forearm stump. METHODS: The female patient was 19 years old, whose right hand and wrist were crushed into pieces by machine at work and her forearm was amputated at the level of 8 cm proximal to the wrist. The second toe of her left foot was transplanted to reconstruct the digit onto the stump of her forearm. Two months after the transplantation, the patient was transferred to the rehabilitation center for further rehabilitation training, which consisted of: training for adaptation to weight bearing, testing and training of sensibility to weight. testing and training for stability of the hand, and testing and training for the controlling function of the reconstructed digit. RESULTS: The transplanted toe survived well. After rehabilitation the reconstructed digit functioned well. In testing the performance under control mandate, the accuracy rate of the electronic artificial hand was 100%. CONCLUSIONS: A 100% accuracy rate of the electronic artificial hand can be achieved by transplantation of the toe onto the stump of the forearm. It provides a useful pathway and an example for improvement of control accuracy of a multiple freedom electronic artificial hand and reduction of false action. PMID- 11882259 TI - The effects of stress-relaxation plate on disorganization and repair of regional bone structure: an experimental study. AB - OBJECTIVE: To study the influence of stress-relaxation plate on disorganization and repair of the cortex beneath the plate. METHODS: A washer made of viscoelastic polyethylene was placed between the screw and the screw hole of conventional stainless rigid plate (RP) to produce a stress-relaxation plate (SRP). Both SRP and RP were applied to osteotomized tibia in 48 New Zealand rabbits. Healing process of the fracture with either SRP or RP fixation (control) was comparatively studied with polarized light microscopy, in situ hybridization of collagen mRNA and immunohistochemical technique from 2 to 36 weeks postoperatively. RESULTS: The study of plated bone remodeling showed that the degree of cortex osteoporosis beneath the plate was similar between the SRP and RP group within 12 weeks postoperatively. In comparison, the disorganization of bone structure in SRP group happened later and milder than that of RP group, and the repair process began at 12 weeks after implantation. As a consequence, the absorption cavities became smaller and the structure of collagen fibers became well oriented along with these changes by polarized light microscopy. In addition to these, the in situ hybridization analysis of collagen genes and the immunohistochemical study of type I, III collagen showed that the osteoblasts lying on the surface of absorption cavities expressed and synthesized type I collagen at 8 to 12 weeks after implantation. From this time on, the changes above became more evident significantly before most of cavities were repaired by 36 weeks. In contrast to the changes in the SRP group, no expression and synthesis of any kind of collagen could be observed during 12 to 36 weeks after implantation in RP group. CONCLUSIONS: Without removal of the bone plate, the SRP fixation not only reduces the degree of plated bone osteoporosis, but also makes the disorganized bone structure restored to normal in terms of the expression and synthesis of type I collagen mRNA of osteoblasts lying on the surface of absorption cavities. PMID- 11882260 TI - Operative treatment of medial rotation contracture of the shoulder caused by obstetric brachial plexus palsy. AB - OBJECTIVE: To introduce an operation of subscapularis slide from its origin and anterior release from its insertion for treatment of medial rotation contracture, subluxation and dislocation of the shoulder caused by obstetric brachial plexus palsy (OBPP). METHODS: Thirty-six cases with medial rotation contracture of the shoulder were diagnosed by measurement of the inferior glenohumeral angle, passive lateral rotation of the shoulder and plain radiographs. Subscapularis slide was performed in 24 cases with simple medial rotation contracture, and anterior release in 12 cases with complex contracture-medial rotation contracture combined with subluxation, dislocation, or other deformities of the shoulder joint. Systems of Mallet scoring and Gilbert grading for the shoulder were used to evaluate the postoperative shoulder function. RESULTS: With follow up for a minimum of six months, 32 cases got apparent gains from operations, accounting for 88.8% of the total operated on. The younger the child was, the better the result. Of 4 cases with no operative effects, 3 had no flexion of the elbow preoperatively, suggesting a poor recovery of the upper trunk of the brachial plexus; the rest one had no repair of the severed subscapularis tendon. CONCLUSIONS: Subscapularis slide and anterior release of the shoulder are effective for treatment of medial rotation contracture as well as its consequence of subluxation and dislocation of the shoulder in OBPP. The operative effect is related to children's age and the recovery extent of the upper trunk of the brachial plexus. PMID- 11882261 TI - Biomechanical principle of arcus plantaris and its clinical application. AB - OBJECTIVE: To analyze the principle mechanism of the arcus plantaris and its clinical application. METHODS: The states of forces sustained by the arcus plantaris were analyzed and calculated according to the mechanism of the quadratic parabolic arch. RESULTS: The aponeurosis plantaris corresponded to the pull rod of the arcus plantaris. The medial and lateral longitudinal arches formed by the pedal bones were stable with the rod, but unstable without the rod. In the latter condition, on loading, the force sustained by the parabolic arch became a force sustained by a simple beam, and the arcus plantaris tended to disappear and to be flattened. Clinically, 240 feet with talipes equinus were treated with triple arthrodesis. In 34 out of the reexamined 156 feet, the aponeurosis plantaris was cut in addition to the triple arthrodesis and was immobilized with cast for 3 months. One or two years later, their arcus plantaris disappeared, pain developed when walking, and some of them walked with the midtarsal joint against the ground. Then, the triple arthrodesis and shortening of the aponeurosis plantaris were applied on 18 cases, and osteotomy of the calcaneus and reconstruction of the aponeurosis plantaris were made on 10 cases and satisfactory effects were obtained. CONCLUSIONS: In order to achieve satisfactory therapeutic effects of the triple arthrodesis, we should reestablish the arcus plantaris and accurately treat the aponeurosis plantaris for the balance of the surrounding muscle force. PMID- 11882262 TI - CT finding is an index in assessment of outcome in patients with diffuse traumatic brain swelling. AB - OBJECTIVE: To assess the relationship between the prognosis of the patients with diffuse traumatic brain swelling (DTBS) and the changes of the ventricles and the cisterns in CT scans. METHODS: The outcome of the patients with DTBS and the changes of the ventricles and the cisterns in CT scans were studied and analyzed in a group of 268 cases. We focused on the changes of the third ventricle and the basal cistern, age and Glasgow Coma Scale (GCS). RESULTS: Of 268 cases, there were changes of the third ventricle and/or the basal cistern in 124, 65 died. In l8 cases, the third ventricle and the basal cistern were both absent and l6 died (88.9%). The third ventricle changed significantly in 59 cases, 33 died (55.9%), while the basal cistern changed in 47 cases and 16 died (34%). Of the 124 patients with changes of the third ventricle and/or the basal cistern, 26 were children, 8 died; 98 adults, 57 died. CONCLUSIONS: For patients with DTBS, the outcome was in direct correlation with the change of the third ventricle and/or the basal cistern, the change of the third ventricle was much more important in assessment of the outcome than that of basal cisterns. There is no significant difference in, the incidence of DTBS between children and adults while the outcome of children is much better than that of adults. The patients with the changes of the third ventricle and the basal cistern accompanied with lower GCS scores have poor outcome. PMID- 11882263 TI - In situ localization of procollagen gene expression on cryosections of undecalcified fracture callus. AB - OBJECTIVE: To analyze the expression of procollagen gene in fracture callus, and to search for the technique of in situ hybridization for undecalcified skeletal tissue. METHODS: In situ hybridization of procollagen gene expression was performed on the undecalcified cryosections of rat fracture callus at 7, 14, and 28 d. RESULTS: The hybridization signals achieved were clear and easy to be localized with high specificity. On the 7th day, the expressions of pro alpha1 (III) in fibroblasts and some chondrocyte-like cells were dominant; and at the end of second week high expression of type-II procollagen mRNA was observed in chondrocytes. At the end of fourth week, the cartilaginous callus was almost all replaced by woven bone tissue, and some type-I procollagen mRNA positive osteoblasts and hypertrophic chondrocytes were found scattering in the woven bone and remnants of cartilaginous callus. CONCLUSIONS: The modified method employed in this study is easier, quicker, and more sensitive with high specificity than the conventional tec hnique for in situ hybridization of procollagen gene expression of decalcified rat fracture callus. The phenomenon of shared phenotype expression, which was demonstrated among cells engaged in fracture healing, indicates an important approach to reveal the mechanism of the origin, differentiation, and orientation of cells. PMID- 11882264 TI - Composite mandibular allografts in canines. AB - OBJECTIVE: To evaluate the feasibility of transplanting composite mandibular allografts to repair large mandibular defects. METHODS: Three composite mandibular trans-plantation models were established. The first model consisted of hemimandible with the attached teeth, muscle and skin,and oral mucosa. The second model was transplanted in the same way with the first one excluding oral mucosa and some teeth, and third one excluding the oral mucosa and all dental crowns. Fourteen transplanting operations were performed in canines. Cyclosporine A and methylprednisone were given for immunosuppression. RESULTS: The composite mandibular organs had an effective and closed return circuit. Transplantation of vascularized allograft of mandibular compound organs was feasible. Two longest time survivors of 67 d and 76 d were in the third model group. Cyclosporine A was successful in suppressing rejection of transplanted composite allograft and prolonging survival time of transplantation models. CONCLUSIONS: The composite mandibular allografts were available with large block of living composite tissue, and helpful in restoration of appearance and function for severe mandibular defects. PMID- 11882265 TI - Bridge tendon graft in no man's land: an experimental study in chickens. AB - OBJECTIVE: To investigate the morphological characteristics of the bridge tendon grafting in no man's land to reconstruct the tendon defect and the effect of passive mobilization on it. METHODS: A 2 cm defect was made in bilateral flexor digitorum profundus tendons of the middle chicken toes, and was then transplanted to the opposite site to serve as a segmental autograft tendon. Postoperatively, passive mobilization of the left and right middle toes began at 5 and 21 d separately. Specimens were studied by light, scanning and transmission electron microscopy at 5, 10, 21 and 35 d. RESULTS: Early repair of the tendon-graft of the left middle toes was made by proliferation and ingrowth of the epitenon cells intermingled with newly-formed collagen fibers. A gliding surface formed at 10 and 21 d. The tendon graft itself played an active role in the repair. In contrast, adhesions obliterated the surface and occupied the space between the tendon graft and surrounding tissues in the right middle toes. CONCLUSIONS: It indicates that the use of the segmental bridge tendon graft in no man's land coupled with early passive motion stimulates an intrisic repair process in both the tendon stump and the autogenous tendon graft and results in a functional healing. PMID- 11882266 TI - The ATP-sensitive K(+) channel and membrane potential in the pathogenesis of vascular hyporeactivity in severe hemorrhagic shock. AB - OBJECTIVE: To elucidate the mechanism of vascular hyporeactivity following severe hemorrhagic shock (HS) by studying the changes of ATP-sensitive potassium channels' (K(ATP)) properties and membrane potential of mesenteric arteriolar smooth muscle cells. METHODS: Single channel currents were studied on cell attached and inside-out patches of enzymatically isolated mesenteric arteriolar smooth muscle cells (ASMCs). Membrane potentials of arteriolar strips and ASMCs were recorded by intracellular membrane potential recording method and confocal microscopy, respectively. RESULTS: K(ATP) channels in ASMCs were activated, which induced smooth muscle hyperpolarization following vascular hyporeactivity in HS. CONCLUSIONS: Hyperpolarizing effect of K(ATP) channel activation plays an important role in low vasoreactivity during severe hemorrhagic shock. PMID- 11882267 TI - Content change of neurofilament protein subunits in experimental brain diffuse axonal injury by lateral head rotation. AB - OBJECTIVE: To explore the content change of neurofilament (NF) protein subunits in the experimental brain diffuse axonal injury (DAI) by lateral head rotation. METHODS: Twenty-four Sprague Dawley (SD) rats were equally divided into three injury groups (2 h, 12 h, and 24 h post injury) and one control group. The models of DAI were made in the injury groups by lateral head rotation. Western blotting technique was used to measure the content of NF68 (a kind of NF protein subunit) in the brainstem tissues among all the injured and control rats. The NF68 immunohistochemical staining was used in another six SD rats in order to observe the morphological changes in DAI. RESULTS: The NF68 content in the brainstem tended to decrease at 2 h post injury, decreased significantly at 12 h and continued its decrease at 24 h. NF56 and NF52, as the breakdown products of NF68, had a tendency to increase at 2-12 h after the injury, and amounted to a significantly higher level at 24 h. Microscopically, there were a lot of swelling neuronal axons in the ventral part of the medullar oblongata at 2 h after the injury. Some axons were disconnected, and axonal retraction balls formed on their proximal end. CONCLUSIONS: There is an occurrence of phosphorolysis within the brainstem in DAI by lateral head rotation. These reactions cause the breakdown of NF68, which results in the decrease of NF68 in content. It suggests that the breakdown of neurofilament protein subunits is an important reason for structur al destroy of neurofilaments in DAI. PMID- 11882269 TI - Epidemiological characteristics and preventive strategies for fall injury. AB - OBJECTIVE: To elucidate the epidemiological characteristics and to define some preventive strategies for fall injury (FI). METHODS: The medical records of patients admitted following a fall from a certain height between August 1996 and July 1997 were analyzed retrospectively. RESULTS: A total of 138 patients were assessed, with a mortality of 31.2%. The male-to-female ratio was 3.5:1. The persons between 20 and 59 years old were the main victims (81.8%), of which 52.2% were related with their work altitude. The remaining adults fall because of, accidents in daily life, suicide attempts, drug abuse, alcohol, or criminal behavior. There were significant differences between the death group and the survival group in the Revised Trauma Score (RTS) and the Injury Severity Score (ISS) value (P<0.05 and P<0.01, respectively). Six children fall from balconies, open windows or roofs. There were significant differences for the height of fall and RTS value in aged group than those in children, adolescents, and adults (P<0.001, 0.005, 0.05; and P<0.05, 0.01, 0.05, respectively). The mortality of FI was significantly correlated to the height of fall (r=0.897, P<0.005). CONCLUSIONS: Male adults are the main victims, especially the workers at high altitudes. The mortality of FI is significantly correlated to the height of fall. The preventive strategies developed through analyzing the risk factors of fall in different age groups might reduce the injuries and deaths following fall. PMID- 11882270 TI - Bile duct injury following blunt abdominal trauma. AB - Bile duct injury caused by abdominal trauma, usually accompanied with injuries of other abdominal organs, is rarely seen. For the reason of its complexity and often delayed diagnosis and treatment, bile duct injury usually leads to severe complications such as abdominal infection. This paper reports a group of 10 cases of bile duct injury treated in our center in the recent decade. PMID- 11882268 TI - Posttraumatic expression of MHC II molecules: an experimental and clinical study. AB - OBJECTIVE: To evaluate the role of MHC II expression in posttrauma immune disturbances and infections. METHODS: In experimental study, peritoneal macrophages were harvested from traumatized mice and treated with GM-CSF, Ia molecules were determined by flow cytometry. In 24 trauma patients, monocyte HLA DR expression was measured by APAAP technique. RESULTS: Marked inhibition of MHC II molecule expression was found in both traumatized mice and patients. In traumatic patients with infection, the inhibition was more severe. Treatment with GM-CSF in mice partially restored the Ia expression. CONCLUSIONS: Inhibition of MHC II molecules plays an important role in postrauma immune disturbances. There is a close relationship between HLA-DR inhibition and clinical infections. PMID- 11882271 TI - Treatment of delayed union and nonunion of femoral shaft fracture with interlocked Grosse-Kempf nail. AB - The failure of using general intramedullary nail or compression plating to treat femoral shaft fracture often causes delayed union or nonunion. Since 1995, we have treated 15 patients with delayed union and nonunion of femoral shaft fracture with interlocked Grosse-Kempf nail. Good results have been obtained. PMID- 11882272 TI - Significance of nitric oxide on the pathogenesis of steroid induced femoral head necrosis. AB - Steroid-induced femoral head necrosis is claimed to be an ischemic femoral head disease. But there is no discussion on the role of nitric oxide (NO) in the idiopathic disease. The concentration of NO indirectly in serum with steroid induced avascular necrosis of femoral head (ANFH) and in controls are studied in this article. PMID- 11882273 TI - CT features of high-voltage electric shock-induced liver injury: a case report. AB - Few reports concerning the CT feature of electric shock-induced liver damages have ever been published. This paper presents such a case treated in our Institute. PMID- 11882274 TI - [Different immune profiles according to the immunological and clinical progression in vertically HIV-infected children]. AB - BACKGROUND: Our goal was to evaluate immunologic profile differences of HIV infected children on antiretroviral treatment (ART). PATIENTS AND METHODDS: We studied 23 HIV-vertically infected children: a) N-A1 group: 10 HIV-infected children in A1 category; b) N-B2 group: 6 HIV-infected children in B2 category, and c) N-C3 group: 7 HIV-infected children in C3 category. We also studied 13 healthy age-matched HIV-negative children as controls. Cell proliferation was evaluated by incorporation of [3H]-Thymidine. The cytokine production in culture was quantified using commercially available specific ELISA assays. T-cell subsets were determined by flow cytometry. RESULTS: Stimulation indexes of PHA, PWM, and anti-CD3+ anti-CD28 in N-A1 group were higher than in N-C3 group. In unstimulated PBMC, TNF-alpha production of HIV-infected children was higher than the control group (p < 0.05). However, in stimulated PBMC, TNF-alpha production in N-B2 and N C3 groups was lower than the control group (p < 0.05). In HIV-infected children, CD8+ CD45RA+ CD62L+ T-cells were significantly lower (p < 0.01) and CD8+ CD45RO+ T-cells were higher (p < 0.05) than the control group. Moreover, in NA-1 group, CD4+ CD45RA+ CD62L+ T-cells were higher, and CD4+ CD45RO+ and CD8+ CD45RO+ T cells were lower, than in N-B2 and N-C3 groups (p < 0.05). On the other hand, CD45RO+, CD45RO+ CD38+, HLA-DR+, CD38+ HLA-DR+ and CD38+ CD4+ and CD8+ T-cells were higher in N-C3 group than the N-A1 and control groups, except for CD4+ CD38+ T-cells. Activated CD8+ T-cells in N-A1 group were higher than in control group (p < 0.01). CONCLUSION: Our data demonstrate that in spite of ART, there still remain important differences in the immunologic status of HIV-infected children depending on the HIV-infection stage. PMID- 11882275 TI - [Mutations of resistance of HIV-1 in previously untreated patients at penitentiary centers of the Autonomous Community of Valencia, Spain. REPRICOVA study]. AB - BACKGROUND: Our purpose was to determine the prevalence of mutations of resistance to nucleoside inhibitors of reverse transcriptase (NIRT) and protease inhibitors (PI) in the HIV-1 genotype of naive infected subjects in the prisons of the Autonomous Community of Valencia, Spain. PATIENTS AND METHOD: Multicentric, descriptive, cross-sectional study of prevalence including a systematic stratified and randomised sampling by centres. Demographic, clinical, virological and immunological data were collected. The HIV gene of protease and transcriptase was studied in peripheral blood plasma samples by means of double PCR amplification and subsequent automatic sequence. Reference: wild strain HXB2. RESULTS: Plasma was obtained from 133 individuals (119 men and 14 women). 117 samples were selected and the rest did not have enough copies for transcription. With regard to NIRT, 7 samples (5.2% of total) showed some mutation of resistance: M41L, D67N, L210W and K219Q, all them secondary to and associated with resistance to zidovudine, abacavir as well as group B multinucleoside resistance. With regard to PI, only one sample showed a primary mutation, M46I, which was associated with resistance to indinavir. Moreover, a further 41 samples were found to express some secondary mutation. CONCLUSIONS: In our series, there was a low number of primary mutations of resistance. These results allow us to exclude the systematic use of resistance tests before an initiation antiretroviral therapy. PMID- 11882277 TI - [Sensitization and occupational asthma in poultry workers]. AB - BACKGROUND: A high prevalence of asthma has been reported in poultry farmers. Our aim was to determine air contaminants in poultry confinement buildings and the prevalence of occupational asthma in these workers. PATIENTS AND METHOD: Spanish poultry farmers who participated in the European Study Prevalence and Risk Factors for Airway Obstruction in Farmers, who worked inside confinement buildings, were enrolled in a cross-sectional study that included the measurement of air contaminants (ammonia, total dust, endotoxin and microorganisms), symptoms, skin testing with common and poultry-related allergens, and a specific bronchial challenge. The Spearman correlation coefficient was used to assess the correlation between air contaminants. RESULTS: 14/15 poultry farmers participated in the study (response index: 93.3%). Concentrations of air contaminants in the confinement buildings [median (interquartile range)] were: 7.5 (5-20) ppm for ammonia, 2.6 (1.8-4.6) mg/m3 for total dust, 137.1 (58.6-243.9) ng/m3 for endotoxin and 7,600 (907-27,750) colony-forming units/m3 * 103 for bacteria. Endotoxin correlated with bacterial concentration (r = 0.69, p < 0.01). 5/14 poultry farmers reported wheezing and/or asthma within the last year (35.7%), and in 2 of them spirometry demonstrated current asthma (14.3%). Specific bronchial provocation confirmed the diagnosis of occupational asthma in one subject with current asthma sensitized to the storage mite Lepidoglyphos destructor (7.1%). CONCLUSIONS: One third of poultry farmers working inside confinement buildings reported wheezing, that is partly attributable to occupational asthma caused by storage mites. PMID- 11882276 TI - [In memorian of Sheila Sherlock]. PMID- 11882279 TI - [Rheumatoid arthritis: Something important has changed]. PMID- 11882278 TI - [Distribution of the serum concentration of beta-carotene, retinol and alpha tocopherol in a representative sample of the adult population of Catalonia (Spain)]. AB - BACKGROUND: Some vitamins (beta-carotene, retinol and alpha-tocopherol) have been used in the prevention of neurovegetative diseases, immunodeficiencies and cancer because of the possible relation with the etiology and physiopathology of these disorders. Nevertheless, the serum values of such micronutrients in the population have not been described. The purpose of this study was to determine the serum concentrations of beta-carotene, retinol and alpha-tocopherol in the Catalan population and to examine the prevalence of deficits. METHOD: We studied 343 individuals from a representative sample of individuals aged 18 to 75 years who had participated in the Nutritional Status of the Catalan Population (Spain) 1992-1993 study. Serum concentrations of beta-carotene, retinol and alpha tocopherol were analysed. RESULTS: The serum concentration of beta-carotene was 0.39 and 0.49 micromol/l in men and women, respectively, and the prevalence of deficits was 62% in men and 54% in women. The serum concentration of retinol was lower in women (1.68 mol/l) than in men (1.98 mol/l) but no deficit was detected for this vitamin. 12% of women aged 18 to 34 years showed a marginal deficit of alpha-tocopherol. CONCLUSION: Our study reveals good nutritional status of retinol and alpha-tocopherol in the sample studied. However, a high prevalence of suboptimal concentrations of beta-carotene was detected, which should be corrected increasing the intake of fruits and vegetables. PMID- 11882280 TI - [When and how to drain a pleural effusion?]. PMID- 11882282 TI - [Hepatitis C and cutaneous polyarteritis nodosa]. PMID- 11882281 TI - [Hematopoietic stem cell transplantation in patients or from donors with hepatitis B or C virus infection:review and recommendations]. PMID- 11882283 TI - [Adrenal crisis due to mitotane therapy]. PMID- 11882284 TI - [Asymtomatic infection by Diphyllobothrium latum in a Spanish 3-year-old child]. PMID- 11882285 TI - Requirement of chromatid cohesion proteins rad21/scc1 and mis4/scc2 for normal spindle-kinetochore interaction in fission yeast. AB - BACKGROUND: Proteins conserved from yeast to human hold two sister chromatids together. The failure to form cohesion in the S phase results in premature separation of chromatids in G2/M. Mitotic kinetochores free from microtubules or the lack of tension are known to activate spindle checkpoint. RESULTS: The loss of chromatid cohesion in fission yeast mutants (mis4-242 and rad21-K1) leads to the activation of Mad2- and Bub1-dependent checkpoint, possibly due to a diminished microtubule-kinetochore interaction. Bub1, a checkpoint kinase, localizes briefly at early mitotic kinetochores in wild-type, whereas the cohesion mutation greatly increases the duration of kinetochore localization. Bub1 is bound to the central centromere region of mitotic cells. These cohesion mutants are hypersensitive to a tubulin poison and are synthetic lethal with dis1 and bir1/cut17, which are defective in microtubule-kinetochore interaction. The formation of specialized centromere chromatin containing CENP-A does not require cohesion. Dominant-negative noncleavable Rad21 fails to activate checkpoint but blocks sister chromatid separation and full spindle elongation in anaphase. CONCLUSIONS: Mis4 and Rad21 (budding yeast Scc2 and Scc1 homologs, respectively) act in establishing the normal spindle-kinetochore interaction in early mitosis and inhibit sister chromatid separation until the cleavage of Rad21 in anaphase. Checkpoint directly or indirectly monitors the states of cohesion in early mitosis. Full spindle extension occurs with unequal nuclear division in cohesion mutants in the absence of Mad2. PMID- 11882286 TI - Marker correspondence, not processing latency, determines temporal binding of visual attributes. AB - BACKGROUND: When simultaneous visual events appear to occur at different times, the discrepancy has generally been ascribed to time differences in neural transmission or cortical processing that lead to asynchronous awareness of the events. RESULTS: We found, however, that an apparent delay of changes in motion direction relative to synchronous color changes occurs only for rapid alternations, and this delay is not accompanied by a difference in reaction time. We also found that perceptual asynchrony depends on the temporal structure of the stimuli (transitions [first-order temporal change] versus turning points [second order temporal change]) rather than the attribute type (color versus motion). CONCLUSIONS: We propose that the perception of the relative time of events is based on the relationship of representations of temporal pattern that we term time markers. We conclude that the perceptual asynchrony effects studied here do not reflect differential neural delays for different attributes; rather, they arise from a faulty correspondence match between color transitions and position transitions (motion), which in turn results from a difficulty in detecting turning points (direction reversals) and a preference for matching markers of the same type. PMID- 11882287 TI - How myxobacteria glide. AB - BACKGROUND: Many microorganisms, including myxobacteria, cyanobacteria, and flexibacteria, move by gliding. Although gliding always describes a slow surface associated translocation in the direction of the cell's long axis, it can result from two very different propulsion mechanisms: social (S) motility and adventurous (A) motility. The force for S motility is generated by retraction of type 4 pili. A motility may be associated with the extrusion of slime, but evidence has been lacking, and how force might be generated has remained an enigma. Recently, nozzle-like structures were discovered in cyanobacteria from which slime emanated at the same rate at which the bacteria moved. This strongly implicates slime extrusion as a propulsion mechanism for gliding. RESULTS: Here we show that similar but smaller nozzle-like structures are found in Myxococcus xanthus and that they are clustered at both cell poles, where one might expect propulsive organelles. Furthermore, light and electron microscopical observations show that slime is secreted in ribbons from the ends of cells. To test whether the slime propulsion hypothesis is physically reasonable, we construct a mathematical model of the slime nozzle to see if it can generate a force sufficient to propel M. xanthus at the observed velocities. The model assumes that the hydration of slime, a cationic polyelectrolyte, is the force-generating mechanism. CONCLUSIONS: The discovery of nozzle-like organelles in various gliding bacteria suggests their role in prokaryotic gliding. Our calculations and our observations of slime trails demonstrate that slime extrusion from such nozzles can account for most of the observed properties of A motile gliding. PMID- 11882288 TI - Cadherin-directed actin assembly: E-cadherin physically associates with the Arp2/3 complex to direct actin assembly in nascent adhesive contacts. AB - Cadherin cell adhesion molecules are major determinants of tissue patterning which function in cooperation with the actin cytoskeleton. In the context of stable adhesion, cadherin/catenin complexes are often envisaged to passively scaffold onto cortical actin filaments. However, cadherins also form dynamic adhesive contacts during wound healing and morphogenesis. Here actin polymerization has been proposed to drive cell surfaces together, although F actin reorganization also occurs as cell contacts mature. The interaction between cadherins and actin is therefore likely to depend on the functional state of adhesion. We sought to analyze the relationship between cadherin homophilic binding and cytoskeletal activity during early cadherin adhesive contacts. Dissecting the specific effect of cadherin ligation alone on actin regulation is difficult in native cell-cell contacts, due to the range of juxtacrine signals that can arise when two cell surfaces adhere. We therefore activated homophilic ligation using a specific functional recombinant protein. We report the first evidence that E-cadherin associates with the Arp2/3 complex actin nucleator and demonstrate that cadherin binding can exert an active, instructive influence on cells to mark sites for actin assembly at the cell surface. PMID- 11882289 TI - Each actin subunit has three nebulin binding sites: implications for steric blocking. AB - Nebulin is a giant protein that spans most of the muscle thin filament. Mutations in nebulin result in myopathies and dystrophies. Nebulin contains approximately 200 copies of approximately 35 residue modules, each believed to contain an actin binding site, organized into seven-module superrepeats. The strong correlation between the number of nebulin modules and the length of skeletal muscle thin filaments in different species suggests that nebulin determines thin filament length. Little information exists about the interactions between intact nebulin and F-actin. More insight has come from working with fragments of nebulin, containing from one to hundreds of actin binding modules. However, the observed stoichiometry of binding between these fragments and actin has ranged from 0.4 to 13 modules per actin subunit. We have used electron microscopy and a novel method of helical image analysis to characterize complexes of F-actin with a nebulin fragment. The fragment binds as an extended structure spanning three actin subunits and binding to different sites on each actin. Muscle regulation involves tropomyosin movement on the surface of actin, with binding in three states. Our results suggest the intriguing possibility that intact nebulin may also be able to occupy three different sites on F-actin. PMID- 11882290 TI - Mks1 in concert with TOR signaling negatively regulates RTG target gene expression in S. cerevisiae. AB - The target of rapamycin (TOR) signaling pathway allows eukaryotic cells to regulate their growth in response to nutritional cues. In S. cerevisiae, TOR controls the expression of genes involved in several nutrient-responsive biosynthetic pathways. In particular, we have demonstrated that TOR negatively regulates a concise cluster of genes (termed RTG target genes) that encode mitochondrial and peroxisomal enzymes required for de novo amino acid biosynthesis. TOR acts in part by regulating the subcellular localization of the Rtg1/Rtg3 transcription factor complex. Nuclear entry of this complex requires the cytoplasmic protein Rtg2, whose precise function has remained ill defined. Here we establish that the likely role of Rtg2 is to antagonize the activity of another protein, Mks1, which we demonstrate is itself a negative regulator of RTG target gene activation. Results of epistasis analyses suggest that Rtg2 and Mks1 act downstream of TOR and upstream of Rtg1 and Rtg3. Moreover, we find that Mks1 phosphorylation responds to TOR as well as to each of the Rtg1-Rtg3 proteins, indicative of complex regulation within this branch of TOR signaling. In addition to RTG target genes, microarray analysis reveals robust expression of lysine biosynthetic genes in mks1Delta cells, which depends on a functional RTG pathway. This latter result provides a molecular explanation for the previous identification of MKS1 as LYS80, a negative regulator of lysine biosynthesis [8]. PMID- 11882292 TI - Speeding up reaction time with invisible stimuli. AB - Normal subjects react more quickly to a pair of visual stimuli than to a stimulus alone. This phenomenon is known as the redundant signal effect (RSE) and represents an example of divided visual attention in which signal processing is carried out in parallel to the advantage of response speed. A most interesting aspect of this phenomenon is that it can occur when one stimulus in a pair cannot be consciously detected because of hemianopia or unilateral extinction resulting from brain damage. Here, we report that a similar dissociation between visual awareness and visually guided behavior is present in normal subjects who show an RSE even when the luminance of one of a pair of stimuli is below detection threshold. The observed RSE cannot be attributed to probability summation because it violates Miller's race inequality and is likely to be related to neural summation between supra- and subthreshold stimuli. Given that a similar implicit RSE is present in hemispherectomy patients, we hypothesize that the site of this summation might be the superior colliculus (SC). PMID- 11882291 TI - Oxygen metabolism causes chromosome breaks and is associated with the neuronal apoptosis observed in DNA double-strand break repair mutants. AB - Cells deficient in a major DNA double-strand break repair pathway (nonhomologous DNA end joining [NHEJ]) have increased spontaneous chromosome breaks; however, the source of these chromosome breaks has remained undefined. Here, we show that the observed spontaneous chromosome breaks are partially suppressed by reducing the cellular oxygen tension. Conversely, elevating the level of reactive oxygen species by overexpressing the antioxidant enzyme superoxide dismutase 1 (SOD1), in a transgenic mouse, increases chromosome breakage. The effect of SOD1 can also be modulated by cellular oxygen tension. The elevated chromosome breakage correlates histologically with a significant increase in the amount of neuronal cell death in Ku86(-/-) SOD1 transgenic embryos over that seen in Ku86(-/-) embryos. Therefore, oxygen metabolism is a major source of the genomic instability observed in NHEJ-deficient cells and, presumably, in all cells. PMID- 11882293 TI - Identification of a novel death domain-containing adaptor molecule for ectodysplasin-A receptor that is mutated in crinkled mice. AB - Hypohydrotic Ectodermal Dysplasia (HED) is a genetic disease seen in humans and mice. It is characterized by loss of hair, sweat glands, and teeth. The predominant X-linked form results from mutations in ectodysplasin-A (EDA), a TNF like ligand. A phenotypically indistinguishable autosomal form of the disease results from mutations in the receptor for EDA (EDAR). EDAR is a NF-kappaB activating, death domain-containing member of the TNF receptor family. crinkled, a distinct autosomal form of HED, was discovered in a mouse strain in which both the ligand (EDA) and receptor (EDAR) were wild-type, suggestive of a disruption further downstream in the signaling pathway. Employing a forward genetic approach, we have cloned crinkled (CR) and find it to encode a novel death domain containing adaptor. crinkled binds EDAR through a homotypic death domain interaction and mediates engagement of the NF-kappaB pathway, possibly by recruiting TRAF2 to the receptor-signaling complex. This is an unprecedented example of naturally occurring mutations in ligand, receptor, or adaptor giving rise to the same phenotypic disease characterized by a defect in the proper development of epidermal appendages. PMID- 11882294 TI - Ectopic B-type cyclin expression induces mitotic cycles in endoreduplicating Arabidopsis trichomes. AB - Cell differentiation is frequently accompanied by a switch from a mitotic division cycle to an endoreduplication cycle. In endoreduplicating cells, DNA synthesis continues in the absence of cell divisions, and it is speculated that endoreduplication represents a shortened mitotic division cycle. In animals, it has been shown that cells switching from mitotic to endoreduplication cycles continue to express factors controlling the G1-S transition, whereas the transcription of mitotic factors controlling the G2-M transition is negatively regulated. It is unknown how the mitotic factors are repressed and what the functional significance of their suppression is. To test the function of two mitotic cyclins in an endoreduplication cycle, we expressed CYCLIN B1;1 and CYCLIN B1;2 in unicellular Arabidopsis trichomes. During wild-type development, trichomes undergo an average of four endoreduplication cycles, leading to a DNA content of approximately 32C. We find that ectopic expression of CYCLIN B1;2, not CYCLIN B1;1, induces mitotic divisions resulting in multicellular trichomes. The CYCLIN B1;2-triggered cell divisions appeared normal with respect to both nuclear division and cytokinesis. We show that CYCLIN B1;2 is misexpressed in the siamese mutant, which also produces multicellular trichomes. Additional overexpression of CYCLIN B1;2 in a siamese mutant background caused a strongly enhanced phenotype. PMID- 11882295 TI - Signal-dependent translocation of transducin, RGS9-1-Gbeta5L complex, and arrestin to detergent-resistant membrane rafts in photoreceptors. AB - Many lines of evidence show that membranes contain microdomains, "lipid rafts", that are different from the rest of the membrane in specific lipid and protein composition. In several biological systems, they were shown to be necessary for trafficking and signal transduction. Here, we investigate if lipid rafts have a role in the regulation of the G protein-mediated pathway underlying vertebrate phototransduction. Photoreceptor membranes contain detergent-resistant membrane (DRM) rafts. Rhodopsin and cGMP phosphodiesterase are found in raft and nonraft portions of the membrane; guanylate cyclase is found exclusively in the raft. Distribution of these proteins does not change in the light or dark. In contrast, the G protein transducin, the RGS9-1-Gbeta5L complex, and the p44 isoform of arrestin undergo dramatic translocation to the raft upon illumination. Phosphorylation of RGS9-1 occurs exclusively in the raft. GTPgammaS or pertussis toxin prevent the light-mediated translocation of transducin and RGS9-1, whereas AlF(minus sign)(4) causes both proteins to move to the raft in the dark. This shows that the Galphat-RGS9-1-Gbeta5L complex has the highest affinity to rafts in the transition state of the GTPase. GTPgammaS binds to transducin at a significantly slower rate in the raft, indicating that this translocation results in a reduced rhodopsin-transducin coupling. Thus, an external signal can rearrange components of a G protein pathway in specific domains of the cell membrane, changing its signaling properties. These findings could reveal a novel mechanism utilized by the cells for regulation of G protein-mediated signal transduction. PMID- 11882296 TI - C. elegans ksr-1 and ksr-2 have both unique and redundant functions and are required for MPK-1 ERK phosphorylation. AB - Kinase Suppressor of Ras (KSR) is a conserved protein that positively regulates Ras signaling and may function as a scaffold for Raf, MEK, and ERK. However, the precise role of KSR is not well understood, and some observations have suggested that KSR might act in a parallel pathway. In C. elegans, ksr-1 is only required for a specific Ras-mediated process (sex myoblast migration) and is a nonessential positive regulator of other Ras-mediated developmental events. We report the existence of a second C. elegans ksr gene, ksr-2, which is required for Ras-mediated signaling during germline meiotic progression and functions redundantly with ksr-1 during development of the excretory system, hermaphrodite vulva, and male spicules. Thus, while the ksr-1 and ksr-2 genes are individually required only for specific Ras-dependent processes, together these two genes appear necessary for most aspects of Ras-mediated signaling in C. elegans. The finding that ksr-2; ksr-1 double mutants have strong ras-like phenotypes and severely reduced or absent levels of diphosphorylated MPK-1 ERK strongly supports models where KSR acts to promote the activation or maintenance of the Raf/MEK/ERK kinase cascade. PMID- 11882297 TI - AID is essential for immunoglobulin V gene conversion in a cultured B cell line. AB - Following productive V gene rearrangement, the functional immunoglobulin genes in the B lymphocytes of man and mouse are subjected to two further types of genetic modification. Class-switch recombination, a region-specific but largely nonhomologous recombination process, leads to a change in constant region of the expressed antibody. Somatic hypermutation introduces multiple single nucleotide substitutions in and around the rearranged V gene segments and underpins affinity maturation. However, in chicken and rabbits (but not man or mouse), an additional mechanism, gene conversion, is a major contributor to V gene diversification. It has been demonstrated recently that both switch recombination and hypermutation are ablated in mice and humans lacking AID, a B cell-specific protein of unknown molecular activity. Here we show that disruption of AID in the DT40 chicken B cell lymphoma leads to a failure to perform immunoglobulin V gene conversion. Thus, AID is required for all three immunoglobulin gene modification programs (gene conversion, hypermutation, and switch recombination) and acts in the initiation or execution of these processes rather than in bringing the B cell to an appropriate stage of differentiation. PMID- 11882298 TI - Triple vaccine fears mask media efforts at balance. PMID- 11882300 TI - Netrins. PMID- 11882301 TI - A new family of putative insulin receptor-like proteins in C. elegans. PMID- 11882302 TI - The functional architecture of attention. PMID- 11882303 TI - Light reception: discovering the clock-eye in mammals. AB - Light is the most reliable environmental signal for adjusting biological clocks to the 24-hour day. Mammals receive this signal exclusively through the eyes, but not just via rods and cones. New evidence has been uncovered for a novel photoreceptor that may be responsible for more than just adjusting the clock. PMID- 11882304 TI - Golgi complex: biogenesis de novo? AB - Whether Golgi biogenesis occurs by self-assembly or around a pre-existing template is currently a matter of debate. Recent studies have shown that Golgi structural proteins are more dynamic than previously thought, suggesting that self-assembly of the Golgi complex may be possible. PMID- 11882305 TI - Cortical plasticity: time for a change. AB - Two recent studies have tested whether synaptic learning rules, inferred earlier from work on cell cultures and brain slices, apply in intact brains. The evidence indicates that they do, and reveals interesting implications for brain development and perceptual learning. PMID- 11882306 TI - Cranial nerve development: placodal neurons ride the crest. AB - Neurons of the vertebrate cranial sensory ganglia arise from both neural crest and a series of ectodermal thickenings termed neurogenic placodes. Recent results lend insight into how these two populations of cells coordinate their development, and subsequently innervate their central target, the hindbrain. PMID- 11882307 TI - T memory cells: quality not quantity. AB - Immunological memory to pathogens is associated with clonal expansion and heightened responsiveness of antigen-specific T cells. Recent work emphasizes that, for efficient protective immunity, qualitative changes in T memory cells are vitally important. PMID- 11882308 TI - Apoptosis: letting slip the dogs of war. AB - Recent studies have shown that, during cell death, the protein Omi is released from the mitochondrial intermembrane space into the cytosol, where it augments caspase-dependent apoptosis by blocking inhibitors and may induce caspase independent cell death via its serine protease activity. PMID- 11882309 TI - Behavioral genetics: a gene for supersociality. AB - In the fire ant, the number of queens per colony is determined by the workers' Gp 9 genotype. This gene has been found to encode an odorant binding protein, which probably influences workers' abilities to recognize queens and regulate their numbers. Remarkably, the same gene seems to control social organization in three other closely related species. PMID- 11882310 TI - ER dislocation: Cdc48p/p97 gets into the AAAct. AB - Misfolded or unassembled proteins present in the lumen of the endoplasmic reticulum are exported to the cytosol and degraded. Recent studies have implicated a complex containing the AAA ATPase Cdc48p/p97 in the export process. PMID- 11882311 TI - Order and disorder in the nucleus. AB - Fluorescence in situ hybridization combined with three-dimensional microscopy has shown that chromosomes are not randomly strewn throughout the nucleus but are in fact fairly well organized, with different loci reproducibly found in different regions of the nucleus. At the same time, increasingly sophisticated methods to track and analyze the movements of specific chromosomal loci in vivo using four dimensional microscopy have revealed that chromatin undergoes extensive Brownian motion. However, the diffusion of interphase chromatin is constrained, implying that chromosomes are physically anchored within the nucleus. This constraint on diffusion is the result of interactions between chromatin and structural elements within the nucleus, such as nuclear pores or the nuclear lamina. The combination of defined positioning with constrained diffusion has a strong impact on interactions between chromosomal loci, and appears to explain the tendency of certain chromosome rearrangements to occur during the development of cancer. PMID- 11882312 TI - HDL and arteriosclerosis: beyond reverse cholesterol transport. AB - The inverse correlation between serum levels of high density lipoprotein (HDL) cholesterol and the risk of coronary heart disease, the protection of susceptible animals from atherosclerosis by transgenic manipulation of HDL metabolism, and several potentially anti-atherogenic in vitro-properties have made HDL metabolism an interesting target for pharmacological intervention in atheroslcerosis. We have previously reviewed the concept of reverse cholesterol transport, which describes both the metabolism and the classic anti-atherogenic function of HDL (Arterioscler. Thromb. Vasc. Biol. 20 2001 13). We here summarize the current understanding of additional biological, potentially anti-atherogenic properties of HDL. HDL inhibits the chemotaxis of monocytes, the adhesion of leukocytes to the endothelium, endothelial dysfunction and apoptosis, LDL oxidation, complement activation, platelet activation and factor X activation but also stimulates the proliferation of endothelial cells and smooth muscle cells, the synthesis of prostacyclin and natriuretic peptide C in endothelial cells, and the activation of proteins C and S. These anti-inflammatory, anti-oxidative, anti-aggregatory, anti-coagulant, and pro-fibrinolytic activities are exerted by different components of HDL, namley apolipoproteins, enzymes, and even specific phospholipids. This complexity further emphasizes that changes in the functionality of HDL rather than changes of plasma HDL-cholesterol levels determine the anti-atherogenicity of therapeutic alterations of HDL metabolism. PMID- 11882313 TI - 3-Hydroxy-3-methyl-glutaryl coenzyme A reductase inhibitors, atorvastatin and simvastatin, induce apoptosis of vascular smooth muscle cells by downregulation of Bcl-2 expression and Rho A prenylation. AB - The mechanism by which 3-hydroxy-3-methyl-glutaryl coenzyme A (HMG-CoA) reductase inhibitors (statins) induce apoptosis in vascular smooth muscle cells (VSMCs) is unknown. In this work, we demonstrate that treatment of VSMCs with simvastatin and atorvastatin inhibited Bcl-2 expression in a time and dose-dependent manner, while Bax expression was not modified. This effect was reversed by mevalonate (100 micromol/l), farnesylpyrophosphate (5 micromol/l) or geranylgeranylpyrophosphate (5 micromol/l), suggesting the involvement of protein prenylation. The treatment of VSMCs with lipophilic statins was associated with decreased prenylation of p-21 Rho A and mevalonate, farnesyl pyrophosphate (F-PP) and geranylgeranyl pyrophosphate (G-PP) reversed prenylation to basal levels. In addition, overexpression of constitutively active Q63L Rho A prevented, at least in part, apoptosis induced by statins and downregulation of Bcl-2. We also investigated the participation of caspases (proteases) in the apoptosis induced by statins. The treatment of VSMCs with lipophilic statins induced activation of the caspase 9, the first caspase of the mitochondrial pathway. Coincubation of VSMCs with the caspase inhibitor ZVAD-fmk (100 micromol/l) significantly inhibited lipophilic statin-induced apoptosis. These findings indicate that the downregulation of Bcl-2 by Rho GTPases mediates statin-induced apoptosis and suggest a new potential mechanism of action for these drugs on the regulation of cell number in the atherosclerotic lesions. PMID- 11882314 TI - Bone marrow chimerism prevents atherosclerosis in arterial walls of mice deficient in apolipoprotein E. AB - OBJECTIVE: apolipoprotein E (apoE) is a key regulator in cholesterol-rich lipoprotein metabolism. Inherited deficiency of this protein results in type III hyperlipoproteinemia in humans. ApoE, especially that derived from macrophages, can efficiently protect against development of atherosclerotic lesion. To use stem cell gene therapy or mini-transplant in treating abnormal lipid metabolism and preventing atherosclerosis, a minimal level of bone marrow chimerism must be determined. METHODS: lethally irradiated apoE deficient mice (12-16 weeks of age) fed on normal chow were transplanted with normal bone marrow cells (C57BL/6.Ly5.1) mixed with those of apoE deficient mice (C57BL/6.Ly5.2) at various ratios. Plasma cholesterol levels were determined every 3 weeks for up to 42 weeks. Areas of atherosclerotic lesion in the aortas were quantified 6 months post-transplant. Plasma apoE was measured by Western blot analysis. RESULTS: bone marrow transplantation (BMT) in apoE (-/-) mice resulted in a detectable level of plasma apoE as determined by Western blot analysis. The plasma cholesterol levels in mice with > or = 60% chimerism were normalized by 6 weeks post-transplant. Mice with < or = 40% chimerism showed significant reductions, but not normalization, in the plasma cholesterol levels even at 42 weeks posttransplant. However, atherosclerotic areas observed in 10%-chimeric mice were significantly smaller than those in control mice (P<0.01). Immunohistochemical studies in 10% chimeric mice revealed foam cells derived from donor marrow (apoE (+/+)) and expressed immunoreactive apoE in the atherosclerotic lesion. The positive signals by Western blot analysis were represented in the plasma of up to 8% of the chimeric mice. CONCLUSION: chimerism of 10%, the minimum level analyzed, was sufficient to reduce the severity of atherosclerosis, although the plasma cholesterol levels were not completely normalized. The results indicate that stem cell gene therapy and mini-transplant may provide possible therapeutic approaches to treat patients with abnormal lipid metabolism and atherosclerosis. PMID- 11882315 TI - Differentiation of adherent human monocytes into macrophages markedly enhances tissue factor protein expression and procoagulant activity. AB - Circulating monocytes as well as resident monocyte-derived macrophages (MDMs) in the atherosclerotic plaque express tissue factor. We tested the hypothesis that differentiation of monocytes into macrophages increases their tissue factor expression. This was done by isolating monocytes from human blood and culturing them for 24 h or allowing their differentiation into macrophages for 8 days before assay. Tissue factor procoagulant activity was assessed by recalcification clotting assay while Western blotting quantitated tissue factor protein. Immunocytochemistry was applied to localize antigenic tissue factor in cells. The results showed that freshly-isolated monocytes expressed low baseline procoagulant activity (0.0004+/-0.0003 ng rhTF/10(6) cells) which increased 100 fold in 24-h cultured monocytes (0.04+/-0.01 ng/10(6) cells, P<0.0001) and was associated with detectable tissue factor protein by Western blotting. Furthermore, MDMs expressed 25-fold higher procoagulant activity (1.0+/-0.5 ng/10(6) cells, P<0.04) and increased tissue factor protein content compared with 24-h monocytes as corroborated by immunocytochemistry. LPS stimulation increased baseline binding activity of transcription factor kappaB three-fold (P<0.03) and 2.6-fold (P<0.09) in monocytes and in macrophages, respectively. The results demonstrate that differentiation of human monocytes into macrophages in culture enhances their tissue factor expression. The observed tissue factor increase that can be further stimulated by LPS may contribute to thrombogenicity of macrophage rich atheroma. Activation of the TF-kappaB transduction pathway suggests its potential role in the transcriptional regulation of TF gene in response to acute stimulation in these cells. This cell-culture model may provide the opportunity to evaluate the effect of various interventions on macrophage procoagulant activity. PMID- 11882316 TI - Anti-atherogenic effects of the acyl-CoA:cholesterol acyltransferase inhibitor, avasimibe (CI-1011), in cultured primary human macrophages. AB - Acyl-CoA:cholesterol acyltransferase (ACAT) inhibitors have been shown to reduce atherosclerotic lesions in animals; however, the mechanism(s) for this effect remains unclear. Therefore, we used cultured primary human monocyte-derived macrophages (HMMs) to examine the effect of the ACAT inhibitor, avasimibe (CI 1011), during foam cell formation and during cholesterol efflux from established foam cells. To examine the effect of CI-1011 on foam cell development, HMMs were incubated with aggregated acetylated LDL (ag-acLDL)+/-CI-1011 for 48 h. Total cholesterol (TC) was 29% lower in HMMs incubated with ag-acLDL and CI-1011 compared with ag-acLDL (P<0.05). To determine if TC reduction was due to reduced ag-acLDL uptake by CI-1011, 125I-acLDL binding at 4 degrees C for 4 h to HMMs preincubated with acLDL or ag-acLDL, CI-1011, acLDL+CI-1011, or ag-acLDL+CI-1011 for 48 h was measured. Specific binding was 40% lower in cells preincubated with acLDL+CI-1011, 52% lower in cells preincubated with ag-acLDL+CI-1011 and 49% lower in cells preincubated with CI-1011 compared with cells preincubated with acLDL (P<0.0003). Because CI-1011 appeared to directly affect acLDL binding, 125I acLDL (3-80 microg protein/ml) binding was done in HMMs preincubated with CI-1011 (0-10 microg/ml) for 48 h. The calculated B(max) decreased in HMMs exposed to increasing concentrations of CI-1011, suggesting that CI-1011 altered scavenger receptor function and/or number. To examine the effects of CI-1011 on cholesterol efflux from established foam cells, we first examined whether CI-1011 was cytotoxic. HMMs were preincubated with ag-acLDL for 24 h, and then radiolabeled with [14C]adenine for 2 h (time zero). The radiolabeled cells were exposed to control RPMI medium or the same medium+HDL, CI-1011, or HDL+CI-1011 for 24 h. The release of [14C]adenine into the medium was not significantly different between cells exposed to RPMI, HDL, CI-1011, or HDL+CI-1011, suggesting that CI-1011 was not cytotoxic. Foam cells exposed to RPMI and CI-1011 (1-10 microg/ml) for 48 h showed time dependent reduction in cellular TC mass, with a corresponding increase in radiolabeled unesterified cholesterol into the medium. We then asked whether CI-1011 enhanced apoE mediated cholesterol efflux. Although cellular apoE increased between 2- and 7-fold in foam cells compared to control macrophages, apoE secreted into the medium was not significantly different between cells exposed to RPMI or CI-1011. Thus, CI-1011 exerted anti-atherogenic effects by reducing TC accumulation, inhibiting acLDL binding, and by limiting lipid storage in HMMs. PMID- 11882317 TI - Cholesterol esters regulate apoB48 secretion in CaCo2 cells. AB - In this study, we investigated the effect of atorvastatin, an HMG-CoA reductase inhibitor and CL277082, an ACAT inhibitor, on apolipoprotein B48 synthesis, degradation and secretion in transformed human intestinal enterocytes (CaCo2 cells). Cells were incubated with atorvastatin or CL277082 in the absence or presence of sterol containing media and pulsed with [S35]-methionine and chased with unlabelled methionine. Concomitantly, the effect of atorvastatin and CL277082 on the relative amount of apoB48 protein in cells and media was also quantified by western blotting using an apoB antibody and enhanced chemiluminescence. Suppression of cholesterol synthesis with atorvastatin did not attenuate the production or secretion of apoB48 from CaCo2 cells under basal conditions. On the other hand, suppression of cholesterol biosynthesis with atorvastatin under stimulatory conditions accelerated the degradation of apoB48 in cells without affecting its synthesis or secretion. There was no effect of exogenous sterols on apoB48 secretion. Taken together, neither endogenous nor exogenous cholesterol appears to acutely modulate apoB48 secretion from intestinal cells. In contrast, inhibition of cholesterol esterification with ACAT inhibitor significantly attenuated apoB48 secretion under basal and stimulatory conditions by a mechanism which enhanced apoB48 degradation. Collectively, our results suggest that in CaCo2 cells, newly synthesized cholesterol ester may be an immediate regulator apoB48 secretion. PMID- 11882318 TI - Ramipril administration to atherosclerotic mice reduces oxidized low-density lipoprotein uptake by their macrophages and blocks the progression of atherosclerosis. AB - Foam cell formation, the hallmark of early atherosclerosis, results from cholesterol accumulation in arterial macrophages. Angiotensin-II stimulates foam cell formation and angiotensin converting enzyme (ACE) inhibitors reduce atherosclerosis in animal models. The goal of the present study was to determine the effect of the ACE inhibitor Ramipril on the progression of atherosclerosis in apolipoprotein-E-deficient (E0) mice with already advanced atherosclerosis. Therefore, 4-month-old atherosclerotic E0 mice were treated with Ramipril for 2 and 4 months and compared to age-matched placebo-treated mice, as well as to control young (4-month-old) non-treated E0 mice, for their atherosclerosis. Histomorphometry showed that Ramipril treatment substantially inhibited atherogenesis as shown by 48 and 72% reduction in lesion size at 6 and 8 months of age, respectively, compared to the lesion size in age-matched placebo-treated mice. Moreover, the size of the atherosclerotic lesions in 6- and 8-month-old Ramipril-treated mice was almost identical to the size of atherosclerosis of the 4-month-old control mice. Moreover, Ramipril treatment of E0 mice, significantly reduced oxidized low-density lipoprotein (Ox-LDL) uptake by their peritoneal macrophages (MPM) by 32%, compared to Ox-LDL uptake by MPM from 6-month-old placebo mice, and even reduced it by 12% in comparison to Ox-LDL uptake by MPM from 4-month-old control mice. A significant decrease in the mRNA levels of the Ox-LDL receptor CD36 by 58% was observed in macrophages from 6-month-old Ramipril treated mice compared to macrophages from the 6-month-old placebo-treated mice. There was even a significant reduction (by 32%) in CD36 mRNA levels in macrophages from the 6-month-old Ramipril-treated mice, compared to the CD36 mRNA levels in macrophages from the 4-month-old control mice. We thus conclude that administration of the ACE inhibitor Ramipril to E0 mice, which already exhibit significant atherosclerosis, blocked the progression of the atherosclerotic lesion build-up, a phenomenon that could be related to Ramipril-induced inhibition of Ox-LDL uptake by macrophages. PMID- 11882319 TI - Balloon catheterization induces arterial expression of new Tenascin-C isoform. AB - Migration of smooth muscle cells (SMCs) across the internal elastic lamina is a key step in the development of atherosclerotic or restenotic plaques. Cell movement is a complex and highly dynamic phenomenon, involving the continuous formation and breakage of attachments with the underlying substratum. Tenascin-C (Tn-C), a counter-adhesive extracellular matrix protein, is comprised of several isoforms with distinct biological activities. Neither the structure nor function of these isoforms in SMCs has been defined. We have used primers and RT-PCR to fully identify Tn-C isoforms expressed by SMCs. Cloning and sequence analysis of the PCR product indicated that SMCs express a Tn-C isoform with only repeats A1 and A2 of fibronectin type III repeats. Using A1A2-specific antibodies, cDNA probes and RNase mapping, we observed that the A1A2 isoform is predominantly expressed by cultured SMCs derived from aorta of newborn rats, and its expression is up-regulated by PDGF-BB. In contrast, the expression of this isoform is markedly down-regulated in the SMCs derived from adult rat aorta. Western and Northern blots of injured rat carotid arteries revealed that the A1A2-isoform is expressed in response to injury. Using cultured SMCs, we found that the recombinant A1A2 protein that was found in the newly discovered Tn-C isoform promotes SMC chemotaxis. We conclude that Tn-C isoforms are expressed in a regulated fashion in vascular system. Our findings suggest a new role of Tn-C isoforms in the remodeling of vascular wall. PMID- 11882320 TI - Induction of calcification in rabbit aortas by high cholesterol diets: roles of calcifiable vesicles in dystrophic calcification. AB - Atherosclerotic calcification may weaken the aorta wall and thereby lead to rupture of the vessel. The mechanism whereby aortas undergo calcification remains unclear. Previous reports in this laboratory showed that, after 2 months of cholesterol-supplemental feeding, an increase in calcifiability of membrane vesicles isolated from rabbit aortas precedes substantial arterial calcification. Further, the mineral was deposited by isolated calcifiable vesicles as an amorphous phase similar to minerals in human aortas at an early stage of atherosclerosis. In the current study, atherosclerotic calcification was induced by exposing rabbits to a 1% cholesterol-rich diet for 3 or 6 months. After 3 months of dietary interventions, atherosclerotic lesions were fully developed. Fatty streaks were evident in areas proximal to the heart and became less frequent in the distal areas. However, calcification was not yet identifiable histologically or by using Fourier transform spectroscopy (FT-IR). After 6 months of high cholesterol treatment, aortas were partially calcified. Histochemical staining for mineral revealed that calcification appeared to occur predominantly in the intimal areas immediately adjacent to the media. Fourier Transform Imaging analysis demonstrated that the mineral deposited in atherosclerotic rabbit aortas was a hydroxyapatite-like phase. To determine whether aorta vesicles play a role in mineral formation in aortas, vesicles were isolated from calcified aortas and then their calcifiability was compared to that in normal vesicles. Interestingly, during the course of vesicle isolation, we found that calcifiable vesicles with much higher calcifiability than normal vesicles could be readily isolated from atherosclerotic aortas simply by suspending minced tissues in PBS. The characteristics of the calcification process and the enzymatic contents of isolated vesicles were similar to those obtained using collagenase digestion. Correlatively, mineral deposited by calcifiable vesicles isolated from the calcified aortas was also of hydroxyapatite-like phases. Altogether, these observations indicate that (1) aortic calcification is a later event during atherogenesis, (2) calcifiable vesicles are loosely bound to the matrices of the lesions as the result of the disease process and (3) similarities in the mineral phases between those in aortas and by vesicles during atherogenesis further support the role of calcifiable vesicles in dystrophic calcification. PMID- 11882321 TI - Identification and expression of scavenger receptor SR-BI in endothelial cells and smooth muscle cells of rat aorta in vitro and in vivo. AB - In this study, we used immunoelectron microscopy to investigate the subcellular localization of scavenger receptor class B type I (SR-BI) in the arterial walls of rats. The expression of SR-BI in cultured endothelial and smooth muscle cells of rat aorta after exposure to high-density lipoprotein (HDL) was also investigated by immunofluorescence microscopy and immunoblotting analysis. A peptide containing residues 495-509 from mouse SR-BI (mSR-BI) plus an NH2 terminal cysteine was coupled to hemocyanin to generate mSR-BI antiserum in rabbits. Reactivity of antiserum against the synthetic peptides was confirmed with an enzyme-linked immunosorbent assay (ELISA). The results showed that SR-BI was specifically localized on the surface of the endothelial cells and smooth muscle cells. SR-BI was also observed in the cytoplasm of smooth muscle cells. Immunoblotting analysis indicated that SR-BI was expressed in the cell membrane. The levels of SR-BI increased gradually from 1 to 3 h and decreased at 24 and 48 h after cholesterol-loaded cells were incubated in the culture medium containing HDL. We conclude that SR-BI, a functional receptor for HDL, is expressed in the aortic endothelial cells as well as in smooth muscle cells. This receptor also responds to the presence of HDL in the culture medium. PMID- 11882322 TI - Attenuation of matrix protein secretion by antisense oligodeoxynucleotides to the cyclin kinase inhibitor p21(Waf1/Cip1). AB - Progressive fibrosis in major organs, including the heart, the kidney and the vascular tree, plays an important role in mediating chronic disease and atherosclerosis. Production of extracellular matrix proteins, in many cases regulated by the growth factor TGF-beta is an essential component of this process. In a parallel manner to TGF-beta, the cyclin kinase inhibitors (CKIs; which are induced by TGF-beta) regulate transit through the cell cycle, and their effect on growth has been shown to be bimodal in the case of vascular smooth muscle (VSM) cells. Using an antisense oligodeoxynucleotide to the CKI p21(Waf1/Cip1), developed in our laboratory and shown to specifically inhibit p21(Waf1/Cip1) protein levels, we asked whether attenuation of the CKI p21(Waf1/Cip1) by transfection of this oligodeoxynucleotide results in the abolition of TGF-beta-mediated growth inhibition and/or diminished matrix protein production and secretion in the presence or absence of TGF-beta. Specific inhibition of p21(Waf1/Cip1) protein with the antisense oligodeoxynucleotide markedly reduces the production and secretion of the matrix proteins fibronectin and laminin, both in the presence and absence of TGF-beta stimulation, in VSM cells as observed by Western blotting of cell lysate and conditioned medium. In addition, TGF-beta-mediated cell growth inhibition, though attenuated by this oligo, is preserved. Due to the relative ease and safety of transfecting antisense oligodeoxynucleotides into VSM, we believe that this work unmasks a potentially powerful technique for inhibition of matrix protein synthesis in VSM and related cell lines, and may lead to new treatment strategies for atherosclerotic as well as other systemic diseases characterized by aberrant matrix protein secretion. PMID- 11882323 TI - Injury induced neointima formation and its inhibition by retrovirus-mediated transfer of nitride oxide synthase gene in an in-vitro human saphenous vein culture model. AB - Human saphenous veins were cultured to characterize neointima formation and feasibility of gene transfer to inhibit the intimal proliferative response to injury. Mechanical injury was introduced by abrading the luminal surface of the vein patch with a sterile cotton bud. Both injured and non-injured vein patches were cultured and transduced with retroviral vectors carrying marker or therapeutic genes. After a 14-day culture, the thickness of the intimal layer of non-injured vein patches reached 90+/-28 microm at the edge and 61+/-22 microm at the center (n=29) from the original 22+/-12 microm at harvest (n=6, P=0.02). Mechanical injury to the intimal surface prior to culture resulted in an exaggerated proliferative response. The intimal thickness of injured vein patches increased from 3.4+/-1 microm right after injury to 128+/-23 microm (n=12, P<0.001) at the edge after 14-day culture. Genes were transduced efficiently into a luminal layer of cultured veins using a pseudotyped murine leukemia viral vector. Transduction of gene encoding nitric oxide synthase resulted in reduction of neointima formation to 33+/-7 microm (n=12) at the edge after 14-day culture compared to 90 microm (P<0.01) seen in untransduced non-injured vein patches. Marker gene transduction did not alter intimal proliferative response or its immunohistochemical profile. The data suggest that cultured vein can be used as a model for studying the effects of injury to blood vessels and to evaluate the effects of candidate therapeutic genes. PMID- 11882324 TI - Production, characterization, and interspecies reactivities of monoclonal antibodies against human class A macrophage scavenger receptors. AB - Class A macrophage scavenger receptor (SR-A) is one of the major receptors of macrophages and plays important roles in atherogenesis and host defense mechanisms. To assess the role of SR-A, monoclonal antibodies were generated by immunizing SR-A-deficient mice with a recombinant protein of human type I SR-A as immunogen. Four antibodies (SRA-C6, SRA-D10, SRA-E5, and SRA-F8) were confirmed to be specific for SR-A by Western blot analysis. In early atherosclerotic lesions, these antibodies recognized scattered macrophages in intima and foamy macrophages in the periphery of atheromatous cores. Interestingly, foamy macrophages in the core lesion were only weakly stained. In other organs, the antibodies recognized tissue macrophages such as alveolar macrophages, Kupffer cells in the liver, red pulp macrophages in the spleen, sinus macrophages in lymph nodes, and interstitial macrophages in various organs. Perivascular macrophages in the brain (Mato cells) were also positive for these antibodies. Freshly isolated blood monocytes were negative; however, they became positive for these antibodies after 1 day in culture. At 3-5 days in culture, the reaction intensity became stronger along their differentiation towards macrophages. Dendritic cells such as interdigitating cells of lymphoid tissues and epidermal Langerhans cells were invariably negative. In the reaction with animal tissues, each antibody showed a unique reaction pattern. Among four antibodies, SRA-E5 recognized SR-A molecules in all animal species examined, including rats and mice. These antibodies will be useful tools for the study of SR-A in atherogenesis and various other pathological conditions in humans and animal species. PMID- 11882325 TI - Effect of diet on adipose tissue and skeletal muscle VLDL receptor and LPL: implications for obesity and hyperlipidemia. AB - This study was designed to examine the effect of a high-fat (primarily saturated), refined-carbohydrate (sucrose) diet (HFS), which is known to induce obesity and hyperlipidemia, on adipose tissue and skeletal muscle lipoprotein lipase (LPL) and very-low density lipoprotein receptor (VLDL-R) protein expressions. Female Fischer rats were placed on either a HFS or a low-fat, complex-carbohydrate (LFCC) diet for 22 months beginning at 2 months of age. After 20 months, a subgroup of the HFS rats were switched to the LFCC diet for 2 months (HFS/LFCC). Body weight, feed efficiency, plasma total cholesterol, VLDL C, low-density lipoprotein cholesterol (LDL-C) and triglyceride (TG) concentrations and LDL-C to high-density lipoprotein cholesterol ratio were all significantly raised by the HFS diet and improved by conversion to the LFCC diet. Adipose tissue heparin-releasable, extractable and total LPL activity expressed per cell were significantly increased in the HFS-fed group. However, LPL protein abundance normalized against total cellular protein was unchanged in the HFS group. This observation is consistent with the presence of adipose tissue hypertrophy. Skeletal muscle LPL protein abundance and heparin-releasable activity were reduced by the HFS diet and improved after switching to the LFCC diet. Both adipose tissue and skeletal muscle VLDL-R protein levels were significantly reduced by the HFS diet and increased after conversion to the LFCC diet. We conclude that an HFS diet induces changes in LPL and VLDL-R in a manner which favors shunting of dietary fat from skeletal muscle to adipose tissue and decreases TG-rich lipoprotein clearance contributing to increased plasma lipids and obesity. Conversion to a LFCC diet can ameliorate the dyslipidemia and tissue changes induced by long-term HFS diet consumption. PMID- 11882327 TI - Effect of anti-tumor necrosis factor-alpha polyclonal antibody on restenosis after balloon angioplasty in a rabbit atherosclerotic model. AB - Inflammation has been postulated to contribute to restenosis after balloon angioplasty. Tumor necrosis factor (TNF)-alpha is a pleiotropic proinflammatory cytokine involved in many features of inflammation. We examined the tissue expression pattern of TNF-alpha and the inflammatory response to arterial injury, and the effects of a goat anti-rabbit-TNF-alpha polyclonal antibody on tissue TNF alpha expression, inflammation and restenosis in a rabbit atherosclerotic model. At different time points following air dessication and subsequent balloon injury, fresh rabbit femoral artery tissues were homogenized and analyzed for TNF-alpha levels by quantitative TNF-alpha bioassay. Rabbits were treated with a goat anti rabbit-TNF-alpha polyclonal antibody, Serum and tissue TNF-alpha neutralization, macrophage infiltration (as an indicator of inflammation), and neointimal areas were determined. Balloon angioplasty increased tissue TNF-alpha expression 100000 fold over baseline, and this increase persisted over 6 days after arterial injury, serum anti-TNF-alpha antibody levels were sufficient to neutralize tissue TNF-alpha activity by 60-75%, macrophage infiltration was suppressed, but did not decrease the neointimal formation. These data indicate that tissue TNF-alpha levels were markedly increased after balloon angioplasty. Anti-TNF-alpha treatment was sufficient to neutralize tissue TNF-alpha activity, reduce inflammation, but did not inhibit neointimal formation following balloon angioplasty in a rabbit atherosclerotic model. PMID- 11882326 TI - Apoptosis of vascular smooth muscle cells is induced by Fas ligand derived from monocytes/macrophage. AB - Fas and its ligand (FasL), are a receptor-ligand pair identified as promoting cell death in several tissues. Apoptosis of vascular smooth muscle cells (VSMCs) in human atherosclerotic plaque may contribute to weakening of the fibrous cap, ultimately resulting in plaque rupture. We investigated the ability of monocytes to induce apoptosis of cultured VSMCs through Fas/FasL pathway. In addition, we examined the association of FasL with apoptosis in human coronary plaques. Both activated monocytes and the supernatant obtained from activated monocytes were able to kill cultured VSMCs. The apoptotic response of VSMCs was almost completely blocked by the caspase inhibitor z-VAD-fmk and was partially blocked by incubation with antagonistic anti-Fas IgG1 which suggests that Fas/FasL system was involved in the induction of cell death. An approximate 30 kDa protein, which represents a cleaved, soluble form of FasL, was identified in culture medium from activated monocytes, but not in culture medium from control, unactivated monocytes. Immunohistochemical analysis of human atherosclerotic coronary lesions showed that FasL is expressed by macrophages, and microvessels in the adventitia as well as in the plaque. Finally, double-staining with terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) and FasL antibody showed that FasL enriched lesions always included a number of TUNEL-positive cells. These data suggest that Fas/FasL pathway can be employed by monocytes/macrophages to induce VSMC apoptosis in the atherosclerotic lesions. PMID- 11882328 TI - Endothelial markers and adhesion molecules in acute ischemic stroke--sequential change and differences in stroke subtype. AB - The progress of a stroke concerns the activation of endothelial cells and platelets. We measured the plasma activities of von Willebrand factor (vWf) and the serum levels of soluble thrombomodulin (sTM) as endothelial markers, and the plasma concentrations of soluble P-selectin (sP-selectin) and soluble E-selectin (sE-selectin) as adhesion molecules during the acute (within 48 h from onset) and subacute (after 1 month from the onset) phases of 52 consecutive patients with acute ischemic stroke and 86 age-matched control subjects. The plasma vWf activities and levels of sP-and sE-selectins in stroke patients were significantly elevated compared with those in controls during both the acute and subacute phases. The serum levels of sTM in stroke patients were significantly higher than those in controls only during the subacute phase. In atherothrombotic infarction, the vWf activities and the levels of sP-selectin, markers for endothelial and platelet activation, remained higher until the subacute phase compared with controls, and the concentrations of sTM, a marker for endothelial injury, were increased during the subacute phase compared with during the acute phase. In lacunar infarction, the levels of sTM and sE-selectin of patients were higher only during the acute phase than controls. These findings suggest that the endothelial cell damage might be maintained until the subacute phase in atherothrombotic infarction, whereas it is remarkable only during the acute phase in lacunar infarction. The evaluation of endothelial markers and adhesion molecules would represent the pathophysiological states of stroke and may provide useful information for the treatment of the ischemic infarction. PMID- 11882329 TI - Homocysteine attenuates hemodynamic responses to nitric oxide in vivo. AB - Homocysteine is a significant but modifiable risk factor for vascular diseases. While several pathological processes may be involved, homocysteine can cause significant endothelial impairment and compromise vascular NO bioactivity. In the present study, we aimed to assess effects of homocysteine on NO-mediated hemodynamic responses in vivo. We created an acute hyperhomocysteinemia model (plasma homocysteine of 65-276 micromol/l) by continuous venous infusion of D,L homocysteine to anaesthetized Sprague-Dawley rats. Vasodilators including NO donors: S-nitrosohomocysteine (SNOHcy), S-nitrosocysteine (SNOCys) and sodium nitroprusside (SNP), the endothelial NO synthase (eNOS) activator: acetylcholine (ACh), and calcium channel blocker: verapamil and nicardipine, were administered by one bolus injection to the homocysteinemic rats. While homocysteine infusion produced no change in the mean femoral arterial blood pressure, each of these vasodilators led to a rapid and substantial dose-dependent fall in blood pressure. Concurrent homocysteine infusion, however, attenuated the blood pressure lowering effects induced by NO donors (P<0.01), but not by the calcium channel blockers. Homocysteine inhibited not only the endothelial-derived NO as stimulated by ACh, but also the bioactivity of exogenously supplied NO by SNOHcy, SNOCys and SNP. Our findings indicate that homocysteine may have an effect on NO bioproduction and bioavailability. Vasodilating efficacy of commonly used NO donors such as nitroglycerine may be seriously compromised by hyperhomocysteinemia, which is common among ischemic heart disease patients. PMID- 11882330 TI - Leukocytes extravasation in acute homocysteinemic rats. AB - Homocysteine may promote atherogenesis and thrombogenesis by enhancing leukocyte endothelium interactions. We explored this hypothesis in an acute hyperhomocysteinemia rat model, which was created by a continuous venous homocysteine infusion (4 ml/h/kg body weight) with 2.5 and 10 mg/ml D,L homocysteine upto 90 min. Venous homocysteine levels were monitored periodically and varied 65-276 micromol/l, a range observed frequently in homocysteinemic and homocystinuric patients. We measured hemodynamic parameters in mesentery by intravital microscopy in rats infused with homocysteine (N=5 for each dose) and saline (N=7). Homocysteine infusion for 90 min did not change the mean carotid arterial blood pressure, velocity of red blood cells and rolling leukocyte flux. However at the dose of 10 mg/ml the venular wall shear rate was reduced to 66-69% of the pre-infusion value (P<0.05). The leukocyte rolling velocity decreased to 78-82% (P<0.05). The number of leukocytes adhering to the venular wall increased 2.4-fold (P<0.05), and the leukocyte extravasation increased 4.7-fold (P<0.001). Each of these effects was time-dependent and homocysteine dose-dependent. But none were observed in saline infused rats. In conclusion, while homocysteine infusion did not change hemodynamic parameters, it significantly enhanced dose dependent leukocyte-endothelium interactions, which may contribute to homocysteine induced endothelial dysfunction. PMID- 11882331 TI - Matrix metalloproteinase expression in the coronary circulation induced by coronary angioplasty. AB - Recent studies have revealed that matrix metalloproteinases (MMPs) play an important role in cardiovascular remodeling by degrading the extracellular matrix. We investigated changes in the expression of MMPs due to percutaneous transluminal coronary angioplasty (PTCA). We studied 47 patients with ischemic heart disease who underwent elective PTCA on isolated stenotic lesion of left coronary arteries. Twelve patients received conventional balloon angioplasty, 14 percutaneous transluminal rotational atherectomy and 21 stent implantation. Blood samples were drawn from the coronary sinus immediately before and after, as well as 4 and 24 h, after PTCA. Plasma levels of MMP-1, MMP-2, tissue inhibitor of MMP (TIMP)-1 and TIMP-2 were measured by enzyme-linked immunosorbent assay. Plasma MMP-2 activity was determined with the digestion of a specific chromogenic peptide substrate. We could observe serial changes in plasma MMP-1 levels in the coronary circulation only in one patient, because MMP-1 levels were lower than the limit of detection in other patients. On the other hand, plasma MMP-2 levels in the coronary sinus were detectable in all subjects and increased significantly 4 and 24 h after PTCA. Plasma TIMP-1 levels also showed significant increases 4 and 24 h after PTCA, whereas TIMP-2 did not show significant changes. Plasma MMP 2/TIMP-2 ratio and MMP-2 activity in the coronary sinus showed significant increases 4 and 24 h after PTCA. A positive correlation was observed between MMP 2 levels in the coronary sinus 4 h after PTCA and late loss index 6 months after PTCA. MMP-2 levels in the coronary sinus blood were significantly higher in patients with late restenosis than in those without restenosis. PTCA induces increases in plasma MMP-2 levels and activity in the coronary circulation, which may contribute to vascular remodeling and late restenosis after PTCA. PMID- 11882332 TI - Nonobstructive aortic valve calcification: a window to significant coronary artery disease. AB - BACKGROUND: Aortic valve calcification without obstruction (AVC) is common in the elderly and is associated with increased cardiovascular mortality and morbidity. We hypothesized that AVC detected by transthoracic echocardiography (TTE) is a marker for significant coronary artery disease in patients undergoing coronary angiography. METHODS: The study group included 388 patients with AVC (259 males, 129 females; mean age 72.2+/-9 years) who underwent coronary angiography for various indications. Data were compared with 320, age- and sex-matched patients without AVC who underwent coronary angiography for the same indications. AVC was detected in TTE as focal areas of increased echogenicity and thickening of the aortic-valve leaflets without restriction in motion. Significant obstructive coronary artery disease was defined as either a > or = 70% reduction of the internal diameter of the left anterior descending, right coronary, or left circumflex artery distribution or a > or = 50% reduction of the internal diameter of the left main coronary artery. Risk factors for atherosclerosis including hypertension, smoking, hypercholesterolemia and diabetes were also investigated. RESULTS: Compared with control group, the AVC group had a higher prevalence of significant coronary artery disease (90 vs. 85%, P=0.019), and a trend for lower frequency of coronary arteries without obstruction (6 vs. 9%, P=0.l1); a trend was also noted for 3-vessel disease (38 vs. 33%, P=0.14). Multivariate analysis identified age (P=0.000l), sex (P=0.000l), hypercholesterolemia (P=0.005) and AVC (P=0.02) as independent predictors for significant coronary artery disease. CONCLUSIONS: There is a significant association between AVC and significant coronary artery disease in patients undergoing coronary angiography. Thus AVC can serve as a window to atherosclerosis of the coronary arteries. These results reinforce a previous observation regarding association between AVC and increased risk of cardiovascular morbidity. PMID- 11882333 TI - Dose-dependent suppression of serum cholesterol by tocotrienol-rich fraction (TRF25) of rice bran in hypercholesterolemic humans. AB - Tocotrienols are effective in lowering serum total and LDL-cholesterol levels by inhibiting the hepatic enzymic activity of beta-hydroxy-beta-methylglutaryl coenzymeA (HMG-CoA) reductase through the post-transcriptional mechanism. alpha Tocopherol, however, has an opposite effect (induces) on this enzyme activity. Since tocotrienols are also converted to tocopherols in vivo, it is necessary not to exceed a certain dose, as this would be counter-productive. The present study demonstrates the effects of various doses of a tocotrienol-rich fraction (TRF25) of stabilized and heated rice bran in hypercholesterolemic human subjects on serum lipid parameters. Ninety (18/group) hypercholesterolemic human subjects participated in this study, which comprised three phases of 35 days each. The subjects were initially placed on the American Heart Association (AHA) Step-1 diet and the effects noted. They were then administered 25, 50, 100, and 200 mg/day of TRF25 while on the restricted (AHA) diet. The results show that a dose of 100 mg/day of TRF25 produce maximum decreases of 20, 25, 14 (P<0.05) and 12%, respectively, in serum total cholesterol, LDL-cholesterol, apolipoprotein B and triglycerides compared with the baseline values, suggesting that a dose of 100 mg/day TRF25 plus AHA Step-1 diet may be the optimal dose for controlling the risk of coronary heart disease in hypercholesterolemic human subjects. PMID- 11882334 TI - Apolipoprotein E genotype and response of lipid levels to postmenopausal estrogen use. AB - The allelic variation of Apolipoprotein E (ApoE) influences serum lipid levels. Postmenopausal estrogen replacement therapy (ERT) has favorable effects on the serum lipid profile. We examined the effect of ApoE genotype on lipid response to ERT in 692 community-dwelling women aged 60 and older. ApoE genotypes were categorized into three groups: ApoE 2 (E2/E2+E2/E3, n=94), ApoE 3 (E3/E3, n=430), and ApoE 4 (E3/E4+E4/E4, n=142). Compared to 497 women not using ERT, 169 women currently using ERT were younger (P=0.01), had lower levels of total cholesterol (TC; P=0.10) and low-density lipoprotein (P<0.001), higher levels of high-density lipoprotein (HDL; P<0.001) and triglycerides (TG; P=0.009), and were more likely to have had a surgical menopause (P<0.001). No significant differences in body mass index, alcohol intake, physical activity, or cigarette smoking were found between current ERT users and nonusers (P>0.10). There was a significant interaction between ApoE 2 and ERT for HDL levels: women with ApoE 2 using ERT had the highest HDL levels, and women with ApoE 2 not using ERT had the lowest HDL levels (P=0.015). The unfavorable effect of ApoE 4 genotype on lipoproteins is not altered by HRT, but ApoE 2 genotype modulates the HDL-ERT association in older women. PMID- 11882335 TI - Unique character and metabolism of high density lipoprotein (HDL) in fetus. AB - Lipid and lipoprotein profiles, and enzymes for the lipid metabolism were compared between cord and adult blood. Consistent with previous reports, the major lipoprotein in cord blood was high-density lipoprotein (HDL), and that in adult blood was low-density lipoprotein (LDL). The level of apolipoprotein E (apo E) in cord blood was almost equivalent to that in adult blood, while other apolipoproteins and lipids were all lower than the adult level. In cord blood, apo E-rich HDL cholesterol represented more than 30% of total HDL cholesterol (around 11% in adult), and the concentration was about twice of that in adult blood. This apo E-rich HDL cholesterol was poorly esterified (E/T 56%) compared with that in adults (93%). The lecithin:cholesterol acyltransferase (LCAT) activity in cord blood was extremely low, while the activity and mass of cholesteryl ester transfer protein (CETP) were higher than those in adult blood. The apo E genotype did not show influences on total cholesterol, LDL cholesterol, total HDL cholesterol, and apo E rich HDL cholesterol levels in cord blood, as opposed to those in adult blood. The association of D442G mutation of the CETP gene with the increased HDL cholesterol in adult blood was not seen in cord blood. Rather, the mutation was associated with low total cholesterol and LDL cholesterol levels in cord blood. These results indicate that, in fetus, the character and metabolism of HDL, especially of apo E-rich HDL cholesterol, are distinct from those in adults. PMID- 11882336 TI - Acute vigorous exercise primes enhanced NO release in human platelets. AB - Activation of platelets by acute vigorous exercise has been demonstrated by various parameters, including an increase in agonist-induced platelet [Ca2+]i levels. However, direct evidence is lacking regarding how acute exercise affects platelet-derived NO. Twenty-three healthy male non-smokers (21-59 years) underwent a symptom-limited treadmill exercise test. Washed platelets were prepared from blood samples obtained before and immediately after exercise. All subjects completed at least Bruce stage 2 and were each negative for ischemia. With a low dose (2 microg/ml) of collagen, NO release from washed platelets, detected by the NO-selective microelectrode, was significantly increased after exercise (pmols/10(8) platelets, before: 0.64+/-0.11, after: 1.03+/-0.18; P<0.005) without changes in aggregation ability. This enhanced NO release was accompanied by increased platelet [Ca2+]i levels (before: 232+/-25, after: 296+/ 37; P<0.01). With a high dose (5 or 10 microg/ml) of collagen, NO release and aggregation were both modestly, but significantly, enhanced after exercise. The exercise-induced enhancement of platelet NO release in response to collagen was also suggested by increase in platelet cyclic guanosine monophosphate accumulation and augmenting effect of N(G)-monomethyl-L-arginine on platelet aggregation. In summary, acute strenuous exercise primes enhanced NO release and may play a protective role against exercise-induced activation of platelets in normal subjects. PMID- 11882337 TI - Humoral markers of inflammation and endothelial dysfunction in relation to adiposity and in vivo insulin action in Pima Indians. AB - Several studies have shown that humoral markers of inflammation and endothelial dysfunction are predictive of macrovascular events, and correlated with indirect measures of adiposity and insulin action, thus providing a possible link between obesity, insulin resistance and atherosclerosis. We examined the relationship between humoral markers of inflammation and endothelial dysfunction and direct measures of adiposity and insulin action in Pima Indians, a population with a very high prevalence of obesity and insulin resistance, but a relatively low propensity for atherosclerotic disease. Fasting plasma concentrations of the inflammatory markers C-reactive protein (CRP), secretory phospholipase A2 (sPLA2) and soluble intercellular adhesion molecule-1 (sICAM-1) and of the endothelial markers E-selectin and von Willebrand factor (vWF) were measured in 32 non diabetic Pima Indians (18 M/14 F, age 27+/-1 years) in whom percent body fat and insulin-stimulated glucose disposal (M) were assessed by DEXA and a hyperinsulinemic clamp, respectively. CRP, sPLA2, and sICAM-1 were all positively correlated with percent body fat (r=0.71, 0.57, and 0.51, all P<0.01). E-selectin and vWF were not correlated with percent body fat, but were negatively correlated with M (r= -0.65 and -0.46, both P<0.001) and positively correlated with CRP (r=0.46, and 0.33, both P<0.05). These findings indicate that humoral markers of inflammation increase with increasing adiposity in Pima Indians whereas humoral markers of endothelial dysfunction increase primarily in proportion to the degree of insulin resistance and inflammation. Thus, obesity and insulin resistance appear to be associated with low-grade inflammation and endothelial dysfunction, respectively, even in an obesity- and diabetes-prone population with relatively low propensity for atherosclerosis. PMID- 11882338 TI - Serum levels of adhesion molecules correlate with insulin resistance. PMID- 11882339 TI - Maturation alters the contribution of potassium channels to resting and 5HT induced tone in small cerebral arteries of the sheep. AB - To address the hypothesis that maturation alters the contribution of K-channels to resting and agonist-induced tone in small cerebral arteries, second branch middle cerebral arteries (approximately 200 microm) were taken from term fetal (139-141 days gestation) and adult sheep, denuded of endothelium, and mounted in myographs. After determination of length-tension relations, the arteries were stretched to 55, 100, and 145% of optimum length. At each level of stretch, contractile responses to 5 mM 4-aminopyridine (4-AP, voltage-sensitive K-channel blocker), 100 nM iberiotoxin (calcium-sensitive K-channel blocker), 10 microM glibenclamide (ATP-sensitive K-channel blocker), or 10 microM Ba(2+) (inward rectifier K-channel blocker) were recorded. In separate experiments, concentration--response relations were determined for 5-HT in the presence and absence of each of the four K-channel blockers at the same concentrations. Both 4 AP and iberiotoxin produced stretch-dependent contractions of greater magnitude in adult (37% for 4-AP and 43% for iberiotoxin at 100% optimum) than in fetal (5% for 4-AP and 7% for iberiotoxin at 100% optimum) arteries. 4-AP also enhanced the pD(2) for 5-HT in adult (from 7.15 to 7.49), but not in fetal, arteries. Conversely, glibenclamide attenuated the pD(2) for 5-HT in fetal (from 7.02 to 6.71), but not in adult, arteries. Iberiotoxin enhanced the pD(2) for 5-HT in both fetal (from 7.05 to 7.51) and adult (from 7.15 to 7.75) arteries. In addition, iberiotoxin enhanced maximum responses to 5-HT (from 59 to 82%) in adult but not fetal arteries. Finally, 4-AP enhanced the maximum responses to 5 HT in both fetal (from 67 to 85%) and adult (from 59 to 79%) arteries. These results indicate that maturation modulates the contribution of K(V), K(Ca), and K(ATP), but not K(IR) channels to basal and/or 5HT-induced cerebrovascular tone, and demonstrate that K(V) and K(Ca) channels are coupled to stretch-sensitive receptors, and that K(V) and K(Ca) limit contractile responses to 5-HT. To the extent that changes in pD(2) values reflect changes in agonist--ligand interactions, the data also suggest that K(V), K(Ca), and K(ATP) channels may possibly influence ligand--receptor binding for 5-HT. PMID- 11882340 TI - Expression of alpha(2)-adrenergic receptor subtypes in prenatal rat spinal cord. AB - The results of molecular cloning have revealed three subtypes of the alpha(2) adrenergic receptors (alpha(2) AR) that have been defined alpha(2)C10 (alpha(2A)), alpha(2)C2 (alpha(2B)) and alpha(2)C4 (alpha(2C)). The differential expression of alpha(2) AR subtypes is affected by developmental factors in rat submandibular gland, lung and brain. In the spinal cord of postnatal and adult rats, alpha(2A) and alpha(2C) AR subtypes are expressed and appear to mediate pain perception. However, the relative expression of alpha(2) AR subtypes in the prenatal spinal cord is unknown. In the present study subtype-specific antibodies and reverse transcription-polymerase chain reaction (RT-PCR) were used to determine the expression and localization of the alpha(2) AR subtypes in sections of embryonic day 14 rat spinal cords and primary cultures of cells isolated from these cords. Spinal cords were removed from day 14 embryos, and were sectioned or used for the preparation of cell cultures. After 9 days in culture, neurons were examined by immunofluorescence microscopy or used for preparation of total RNA. In both intact spinal cords and isolated cells, positive immunoreactivity was detected with antibodies against alpha(2A) and alpha(2B) subtypes, but not with antibodies against the alpha(2C) subtype. Using a dual-labeling approach, anti alpha(2A) and anti-alpha(2B) immunoreactivity was present on the same population of neurons. RT-PCR results were consistent with immunofluorescence studies, and showed that mRNA encoding the alpha(2A) and alpha(2B) subtypes was present in total RNA prepared from primary cultures of rat spinal cord neurons. In contrast to spinal cords of postnatal or adult rats that express alpha(2A) and alpha(2C) AR subtypes on different neurons, prenatal spinal cords contain alpha(2A) and alpha(2B) AR subtypes, and these two subtypes appear to be co-expressed in the same cells. PMID- 11882341 TI - 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine has a transient proliferative effect on PC12h cells and nerve growth factor additively promotes this effect: possible involvement of distinct mechanisms of activation of MAP kinase family proteins. AB - 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) is a potent inducer of cell death, producing reactive oxygen species (ROS) and causing apoptosis in PC12h cells at 1 mM [Shimoke et al., J. Neurosci. Res. 63 (2001) 402-409]. We showed here that MPTP also had a weak proliferative effect on PC12h at 500 microM when treated for 24 h. The proliferative effect was additive within 24 h cells when nerve growth factor (NGF) was present in the culture medium, but NGF promoted cell differentiation 2 or 3 days after. Use of PD98059, a specific inhibitor of MEK1 located upstream of extracellular signal-regulated kinases (ERKs), revealed that the NGF- and MPTP-induced proliferative effect depends on the MEK1 pathway because PD98059 diminished the proliferation completely, and interestingly, NGF and MPTP promoted sustained activation of ERKs. Moreover, we observed that MPTP increased the activity of p38 MAPK but not c-jun N-terminal kinase (JNK) in 30 min. We also observed that SB203580, a specific inhibitor of p38 MAPK, decreased cell viability. These results suggest that NGF and MPTP cooperate to promote acute cell proliferation via the sustained ERKs and the p38 MAPK pathway within 24 h in PC12h cells. PMID- 11882342 TI - Ethanol-induced apoptotic neurodegeneration in the developing C57BL/6 mouse brain. AB - Recent studies have shown that administration of ethanol to infant rats during the synaptogenesis period (first 2 weeks after birth), triggers extensive apoptotic neurodegeneration throughout many regions of the developing brain. While synaptogenesis is largely a postnatal phenomenon in rats, it occurs prenatally (last trimester of pregnancy) in humans. Recent evidence strongly supports the interpretation that ethanol exerts its apoptogenic action by a dual mechanism--blockade of NMDA glutamate receptors and hyperactivation of GABA(A) receptors. These findings in immature rats represent a significant advance in the fetal alcohol research field, in that previous in vivo animal studies had not demonstrated an apoptogenic action of ethanol, had not documented ethanol-induced cell loss from more than a very few brain regions and had not provided penetrating insight into the mechanisms underlying ethanol's neurotoxic action. To add to the mechanistic insights recently gained, it would be desirable to examine gene-regulated aspects of ethanol-induced apoptotic neurodegeneration, using genetically altered strains of mice. The feasibility of such research must first be established by demonstrating that appropriate mouse strains are sensitive to this neurotoxic mechanism. In the present study, we demonstrate that mice of the C57BL/6 strain, a strain frequently used in transgenic and gene deletion research, are exquisitely sensitive to the mechanism by which ethanol induces apoptotic neurodegeneration during the synaptogenesis period of development. PMID- 11882343 TI - Oxidative stress and dopamine deficiency in a genetic mouse model of Lesch-Nyhan disease. AB - Lesch-Nyhan disease, a neurogenetic disorder caused by congenital deficiency of the purine salvage enzyme hypoxanthine guanine phosphoribosyl transferase, is associated with a prominent loss of striatal dopamine. The current studies address the hypothesis that oxidant stress causes damage or dysfunction of nigrostriatal dopamine neurons in a knockout mouse model of the disease, by assessing several markers of oxidative damage and free radical scavenging systems. Some of these measures provided evidence for an increase in oxidative stress in the mutant mice (aconitase activity, oxidized glutathione, and lipid peroxides), but others did not (superoxide dismutase, protein thiol content, carbonyl protein content, total glutathione, glutathione peroxidase, catalase, and thiobarbituric reducing substances). Immunolocalization of heme-oxygenase 1 provided no evidence for oxidative stress restricted to specific elements of the striatum or midbrain in the mutants. Striatal dopamine systems of the mutant mice were more vulnerable to a challenge with the neurotoxin 6-hydroxydopamine, but they were not protected by cross-breeding the mutants with transgenic mice over expressing superoxide dismutase. Overall, these data provide evidence for increased oxidative stress, but the failure to protect the knockout mice by over expressing SOD1 argues that oxidative stress is not the sole process responsible for the loss of striatal dopamine. PMID- 11882344 TI - Immediate-early gene expression in concurrent prenatal ethanol- and/or cocaine exposed rat pups: intrauterine differences in cocaine levels and Fos expression. AB - Concurrent use of cocaine and ethanol is a common mode of abuse. Cocaine and ethanol have distinctive pharmacologies but both have been shown to cause uterine vasoconstriction and fetal hypoxia. We developed a paradigm of chronic ethanol exposure via liquid diet coupled with binge cocaine exposure on the last day of gestation. Lipton et al. demonstrated unequal segregation of cocaine in rat fetuses as a function of proximal-distal location in the uterus, indicating a differential vasoconstriction of the two main arteries supplying the uterus in rats receiving cocaine. By performing C-sections after exposure to cocaine, we were able to measure the cocaine content and immediate-early gene (IEG) induction in the brains of fetuses according to their intrauterine position and assess the potentially vasoconstrictive effect of ethanol. HPLC analysis of fetal brains exposed to cocaine supported the study of Lipton et al.: fetuses from the proximal (lower) end of the uterus had more cocaine than fetuses from the distal (upper) end. Concurrent ethanol decreased the amount of cocaine reaching the fetuses and diminished the proximal-distal gradient. There were increased numbers of Fos-immunoreactive cells in fetuses exposed to both ethanol and cocaine compared to cocaine binge only. Additionally, the gradient of c-fos induction observed as a function of intrauterine position in cocaine-treated rats was in the opposite direction: most distal fetuses generally had the most Fos immunoreactive cells. These results indicate that IEG induction in fetal brains exposed to cocaine and ethanol may be more related to hypoxic consequences of prenatal drug exposure. PMID- 11882345 TI - Chlorpyrifos targets developing glia: effects on glial fibrillary acidic protein. AB - The organophosphate pesticide, chlorpyrifos (CPF), is a developmental neurotoxicant. In cell cultures, CPF affects gliotypic cells to a greater extent than neuronotypic cells, suggesting that glial development is a specific target. We administered CPF to developing rats and examined the levels of glial fibrillary acidic protein (GFAP), an astrocytic marker. Prenatal CPF exposure (gestational days 17-20) elicited an increase in GFAP levels in fetal brain, but the effect was seen only at high doses that elicited maternal and fetal systemic toxicity. Early postnatal (PN) CPF treatment (PN1-4) elicited effects only in the cerebellum of male rats; GFAP was suppressed initially (PN5) and showed a rebound elevation (PN10) before returning to normal values by PN30. In contrast, when we administered CPF during the peak of gliogenesis and glial cell differentiation (PN11-14), GFAP was initially decreased across all brain regions and in both sexes; in males, subsequent elevations were seen on PN30, with the largest effect in the striatum; females also showed an increase in striatal GFAP. Our results indicate that CPF disrupts the pattern of glial development in vivo, with the maximum effect corresponding to the peak period of gliogenesis and glial cell differentiation. As glia are responsible for axonal guidance, synaptogenesis and neuronal nutrition, glial targeting suggests that these late-occurring developmental processes are vulnerable to CPF, extending the critical period for susceptibility into stages of synaptic plasticity, myelination, and architectural modeling of the developing brain. PMID- 11882346 TI - Functional alterations in CNS catecholamine systems in adolescence and adulthood after neonatal chlorpyrifos exposure. AB - Chlorpyrifos (CPF), one of the most widely used pesticides, is a neurobehavioral teratogen in animals. We administered CPF to neonatal rats on postnatal days (PN) 1-4 (1 mg/kg) or PN11-14 (5 mg/kg), regimens devoid of overt systemic toxicity. We then examined the impact on catecholaminergic systems in adolescence (PN30) and adulthood (PN60), assessing basal neurotransmitter content and transmitter utilization rates (turnover) in brain regions comprising the major noradrenergic and dopaminergic projections. Although CPF had only sporadic effects on basal norepinephrine and dopamine content, it profoundly suppressed norepinephrine turnover across multiple regions, indicative of net reductions in presynaptic activity. Dopamine turnover showed less consistent effects, with subnormal turnover in some regions and activation in others. We also evaluated whether CPF exposure altered the ability of catecholamine systems to respond to acute cholinergic stimulation, elicited by administration of a single challenge dose of nicotine. In the normal brain, nicotine increases the utilization of norepinephrine and dopamine. With only a few exceptions, animals receiving neonatal CPF exposure showed lasting desensitization of the nicotine response; not only was the activation by nicotine blunted in the CPF group, but in some regions the nicotine response was reversed, eliciting a reduction in transmitter turnover. These results indicate that neonatal CPF exposure produces widespread deficiencies in catecholaminergic synaptic function that persist into adulthood, and that are best revealed by dynamic measures of synaptic activity and responsiveness, as opposed to static markers like basal transmitter levels. The effects seen here are likely to contribute to alterations in behavioral performance that persist or emerge long after the termination of CPF exposure. PMID- 11882347 TI - Perinatal exposure to environmental tobacco smoke upregulates nicotinic cholinergic receptors in monkey brain. AB - In humans, perinatal exposure to environmental tobacco smoke (ETS) is associated with neurobehavioral deficits. In the current study, we exposed Rhesus monkeys to ETS in late gestation and in the early neonatal period, and examined changes in neurotransmitter receptors in the brainstem and caudal portion of the cerebral cortex. Nicotinic acetylcholine receptors were markedly upregulated and the effect was selective in that there were no changes in m(2)-muscarinic acetylcholine receptors or in beta-adrenergic receptors. Nicotinic receptor upregulation is indicative of chronic cell stimulation by nicotine, and is a hallmark of nicotine-induced neuroteratogenesis. These results indicate that perinatal ETS exposes the fetus and neonate to quantities of nicotine that are sufficient to alter brain development. PMID- 11882348 TI - Fumonisin B(1)-induced alterations in cytokine expression and apoptosis signaling genes in mouse liver and kidney after an acute exposure. AB - Fumonisin B(1) (FB(1)), a carcinogenic mycotoxin produced primarily by fungus Fusarium verticillioides in corn, causes several fatal animal diseases. In mice, liver is the primary site of its toxicity. Our previous study showed that maximum induction of interferon gamma (IFNgamma) and tumor necrosis factor alpha (TNFalpha) was observed at 4 and 8 h, respectively, after an acute po FB(1) treatment. To further investigate the time-related induction of other cytokines and genes involved in apoptosis signaling, male BALB/c mice were administered orally with either saline or 25 mg/kg of FB(1) and sampled 4 or 8 h after treatment. Expression of various genes was analyzed by ribonuclease protection assay. FB(1) treatment caused increased expression of TNFalpha and interleukin (IL)-1beta in both liver and kidney, whereas IL-1alpha and IL-1 receptor antagonist (IL-1Ra) expression was induced only in the liver. Expression of TNFalpha signaling molecules, TNF receptor 55 and receptor interacting protein, was increased in liver and kidney after FB(1) treatment. Caspase 8 expression was increased only in liver with no changes in kidney with FB(1). FB(1) treatment induced expression of Fas in liver and kidney with no alterations in Fas signaling molecules, Fas ligand, Fas-associated death domain and Fas-associated protein factor. Treatment of mice with FB(1) increased the expression of B-Myc, c Myc and Max, oncogenic transcription factors in the kidney. FB(1) toxicity caused induction of cytokine network in liver with involvement of TNFalpha signaling pathway. Increased expression of caspase 8 involved in the TNFalpha signaling pathway may contribute to the apoptosis, whereas IL-1Ra induction could contribute to the proliferating effects observed in FB(1) toxicity. PMID- 11882349 TI - Relationship between cadmium concentration in rice and renal dysfunction in individual subjects of the Jinzu River basin determined using a logistic regression analysis. AB - We investigated the association between the Cd concentration in rice and renal dysfunction in individuals living in the Cd-polluted Jinzu River basin, using a logistic regression analysis. In the cases of logistic regression analysis for people (1) who had either resided in the present hamlet since birth or who had moved there from a non-polluted area and for those (2) who had resided in the present hamlet since birth, except for glucosuria in males, all partial correlation coefficients between the Cd concentration in rice and occurrence of abnormal urinary findings were statistically significant in both sexes. The allowable level of Cd concentration in rice was calculated by substituting the abnormality rates of urinary findings of the controls in the 40-49, 50-59 and 60 69 year age groups into the logistic regression formula for people (3). The value for subjects aged 50 years was 0.13 and 0.17 ppm for males and females, respectively, with regard to proteinuria and 0.15 and 0.10 ppm for males and females, respectively, with regard to proteinuria+glucosuria. PMID- 11882350 TI - Fusarenon-X induced apoptosis in HL-60 cells depends on caspase activation and cytochrome c release. AB - Fusarenon-X (FX), a trichothecene mycotoxin, is well known to be cytotoxic to mammalian cells. Our previous study revealed that FX induced apoptosis in mouse thymocytes both in vivo and in vitro. We investigated the mode of apoptosis induced by FX using HL-60 cell culture. When FX at a final concentration of 0.5 microg/ml was added, cell degradation was observed 5 h after exposure, and most of the cells had fallen into apoptosis 24 h after exposure. DNA fragmentation into 180-bp multimers was observed 5 h after exposure, and its dose-dependency was clear in the cells treated with 0.1 microg/ml and higher doses. The percentage of apoptotic cells (sub-G(0) population) increased dose- and time dependently after exposure, when analyzed using flow cytometry. The activities of caspase-3, -8, and -9 were elevated within 2 h by exposure to FX. DNA fragmentation and an increase in the apoptotic population were abrogated by pre treating the cells with broad-spectrum caspase inhibitors Z-VAD-fmk or Z-Asp CH(2)-DCB. Cytochrome c release from mitochondria to cytoplasm was observed clearly, and this release occurred caspase-independently. These findings suggest that FX induces apoptosis in HL-60 cells by stimulating cytochrome c release followed by its downstream events including the activation of multiple caspases. PMID- 11882351 TI - Characterization of 2-amino-4,5-dichlorophenol (2A45CP) in vitro toxicity in renal cortical slices from male Fischer 344 rats. AB - 2-Amino-4,5-dichlorophenol (2A45CP) is a major, aromatic ring hydroxylated metabolite of the renal toxicant, 3,4-dichloroaniline. 3,4-Dichloroaniline is nephrotoxic with primary damage located to the proximal tubules. The purpose of this study was to first characterize the in vitro toxicity of 2A45CP in renal cortical slices. Second, the effect of antioxidants and sulfhydryl containing agents on the severity of 2A45CP toxicity was explored since part of the mechanism of toxicity for aminophenols may involve redox cycling. Renal tissue was isolated from male Fischer 344 rats (190--220 g). Renal slices were rinsed three times for 3 min each in 5-ml Krebs buffer. Tissues were then incubated for 90--120 min with varying concentrations of 2A45CP between 0 and 0.5 mM. In a separate series of experiments, the slices (50--100 mg) were preincubated for 30 min with 1 mM dithiothreitol (DTT), 1 mM glutathione (GSH) or 2 mM ascorbic acid prior to exposure to 0, 0.05, 0.1 or 0.25 mM 2A45CP. 2A45CP produced a concentration and time dependent increase in LDH leakage from renal cortical slices. Total glutathione levels were diminished by 0.5 mM 2A45CP within 30 min. Renal slices incubated for 60 and 120 min with 0.05 and 0.1 mM 2A45CP had lower malondialdehyde levels than control. Pretreatment with DTT did not alter 2A45CP toxicity. Pretreatment of renal cortical slices with GSH or ascorbic acid reduced 2A45CP toxicity. These findings indicate that 2A45CP is directly toxic to renal cortical slices and that cytotoxicity is at least partially mediated by a reactive intermediate. PMID- 11882352 TI - Effects of src-deficiency on the expression of in vivo toxicity of TCDD in a strain of c-src knockout mice procured through six generations of backcrossings to C57BL/6 mice. AB - The effect of TCDD was studied in c-src-deficient C57BL6-src(tm1sor) (N6 src -/- and -/+) mice, and their wild-type littermate mice (N6 src +/+). The former was created from the original strain of B6, 129-src(tm1sor) mice through six generations of backcrossings with C57BL6 mice. The results of a high dose TCDD toxicity tests in male mice indicated that N6 src-/+ mice were significantly less responsive to the toxic action of TCDD (115 microg/kg single i.p. injection) than N6 src+/+ mice in terms of reduced % body weight gain, the increase in the liver to body weight ratio, and the decrease in the adipose tissue to liver weight ratio and in the weight of pancreas. To understand the cause for these differential effects of TCDD we studied TCDD-induced changes in several biochemical parameters at day 10 and found that most drastically affected ones were glycogen depletion and phosphoenolpyruvate carboxykinase (PEPCK) downregulation. In addition, the degree of triglyceride accumulation in liver was less pronounced in N6-/+ than in N6+/+ mice. These findings suggest that the absence of c-src expression indeed affects the development of selected, TCDD induced toxic endpoints that are related to wasting syndrome. PMID- 11882353 TI - Effect of melatonin on the production of microsomal hydrogen peroxide and cytochrome P-450 content in rat treated with aflatoxin B(1). AB - Aflatoxin B(1) (AFB(1)) is a food contaminant fungal toxin that has been implicated as a causative agent in human hepatic and extrahepatic carcinogenesis. In this study we went on to show the effect of melatonin as a free radical scavenger on the production of microsomal hydrogen peroxide (H(2)O(2)) during the metabolic activation AFB(1). The production of microsomal H(2)O(2) in vitro during the metabolic activation of different chemical carcinogens has been reported previously. We also studied the effect of melatonin on the cytochrome P 450 content as a major microsomal monooxygenase isoenzymes system in rat liver responsible for the metabolic activation of AFB(1). The amounts of H(2)O(2) and cytochrome P-450 contents in rat treated with melatonin (0.2 mg/kg BW) and/or AFB(1) (0.2 mg/kg BW) at various time intervals has been measured. Animals treated with melatonin exhibited markedly inhibition in the amounts of H(2)O(2) after 1, 3, and 6 h. The highest level of inhibition (3.0 nmol H(2)O(2)/mg protein) was detected after 6 h. However, cytochrome P-450 contents were also decreased after the same period of time. The highest level of inhibition (2.1 nmol/mg protein) was detected after 3 h of injection. A pronounced augmentation of H(2)O(2) production was observed in rat treated with AFB(1) only. The highest level of H(2)O(2) (100 nmol/mg protein) was measured after 1 h. Cytochrome P-450 contents were also decreased in response to AFB(1) injection over the same time intervals. Contrary data was detected in animals received both AFB(1) and melatonin. The generation of H(2)O(2) was inhibited by melatonin after 1, 3 and 6 h. The highest level of inhibition (44.2 nmol/mg protein) was observed after 6 h. Finally, these data suggested that melatonin as a free radical scavenger inhibited the microsomal production of H(2)O(2) in rat treated with AFB(1). PMID- 11882354 TI - Free radical scavenging activity of red ginseng aqueous extracts. AB - This study was performed to investigate the free radical scavenging activity of Panax red ginseng C.A. Meyer aqueous extract on 1,1-dipheny-2-picrylhydrazyl (DPPH), carbon-centered radical, hydroxyl and superoxide radicals using Electron Spin Resonance (ESR) spectrometer and spin-trapping techniques. Two different Red ginseng aqueous extracts prepared by boiling water or room temperature extraction exhibited no significant difference in free radical scavenging activity. Ginseng extracts completely eliminated DPPH radical at 2 mg/ml. About 0.5 mg/ml ginseng extracts quenched 80% carbon-centered free radicals generated from 2,2'-azobis(2 amidinopropane) dihydrochloride (AAPH). Hydroxyl radical and superoxide radical were generated by UV irradiation and trapped by 5,5-dimethyl-l-pyrroline-N oxide (DMPO). Ginseng extracts scavenged 40% of hydroxyl radical at 0.1 mg/ml. Two mg/ml ginseng extracts completely scavenged superoxide radical. Ginseng extracts did not scavenge nitric oxide. The ESR data demonstrate that red ginseng aqueous extract is not a strong free radical scavenger. PMID- 11882355 TI - Granulocytes from patients with paroxysmal nocturnal hemoglobinuria and normal individuals have the same sensitivity to spontaneous apoptosis. AB - OBJECTIVE: The aim of this study was to determine whether granulocytes from patients with paroxysmal nocturnal hemoglobinuria (PNH) are more or less intrinsically sensitive to spontaneous apoptosis than granulocytes from healthy individuals. Resistance to apoptosis has been suggested as an explanation for the proliferation or selection of PNH clones. PATIENTS AND METHODS: Peripheral blood granulocytes from five patients with PNH, five patients with myelodysplastic syndrome (MDS), and five healthy volunteers were cultured in the absence of serum. Spontaneous apoptosis of the granulocytes was assessed every 6 hours by flow cytometry. The expression levels of CD16b, CD95, and granulocyte-macrophage colony-stimulating factor (GM-CSF) receptor also were studied by flow cytometry, and caspase-3 activity was measured by fluorometry. RESULTS: There were no significant differences in the proportion or absolute numbers of apoptotic and apoptotic/dead granulocytes between the cells from PNH patients and healthy individuals, whereas those from MDS patients showed significantly lower frequencies of apoptotic granulocytes compared with normal controls. The proportion of CD16b(-) granulocytes was not significantly different among the three groups during in vitro culture. CD95 and GM-CSF receptor was not significantly increased in cultured granulocytes or noncultured granulocytes from, respectively, patients with PNH and normal controls. Caspase-3 activity significantly decreased in cultured granulocytes from MDS patients, but not in granulocytes from PNH patients. CONCLUSIONS: Granulocytes from PNH patients did not display a reduced sensitivity to spontaneous apoptosis, suggesting that the apoptosis of blood cells in PNH may not be an important factor in proliferation or selection of PNH clones. These findings are in agreement with the normal lifespan of granulocytes in vivo. PMID- 11882356 TI - Upregulation of natural killer cells functions underlies the efficacy of intratumorally injected dendritic cells engineered to produce interleukin-12. AB - OBJECTIVE: Injection of dendritic cells (DC) engineered with recombinant adenoviral vectors to produce interleukin-12 (IL-12) inside experimental murine tumors frequently achieves complete regressions. In such a system the function of CD8(+) T cells has been shown to be an absolute requirement, in contrast to observations made upon depletion of CD4(+) T cells, which minimally affected the outcome. The aim of this work was to study the possible involvement of natural killer (NK) cells in this setting. MATERIALS, METHODS, AND RESULTS: Depletions with anti-AsialoGM1 antiserum showed only a small decrease in the proportion of complete regressions obtained that correlated with induction of NK activities in lymphatic tissues into which DC migrate, whereas combined depletions of CD4(+) and NK cells completely eliminated the antitumor effects. Likewise in vivo neutralization of interferon-gamma (IFN-gamma) also eliminated those therapeutic effects. Trying to define the cellular role played by NK cells in vivo, it was observed that injection of cultured DC inside the spleen of T- and B-cell deficient (Rag1(-/-)) mice induced upregulation of NK activity only if DC had been adenovirally engineered to produce IL-12. In addition, identically transfected fibroblasts also activated NK cells, indicating that IL-12 transfection was the unique requirement. Equivalent human DC only activated in vitro the cytolytic and cytokine-secreting functions of autologous NK cells if transfected to express human IL-12. CONCLUSIONS: Overall, these results point out an important role played by NK cell activation in the potent immunotherapeutic effects elicited by intratumoral injection of IL-12--secreting DC and that NK activation under these conditions is mainly, if not only, dependent on IL-12. PMID- 11882357 TI - Difference in CD22 molecules in human B cells and basophils. AB - OBJECTIVE: CD22 is believed to be restricted to normal and neoplastic B cells. Human basophils were found to express CD22 molecules. Among the antibodies against CD22, Leu14, which recognized the ligand binding domain, reacted to basophils, and B3 and 4KB128, which recognized the amino terminus side and carboxy terminus side of the ligand binding epitope, respectively, did not. To clarify the difference of CD22 antigenicity in human B cells and basophils, we investigated RNA sequence and structures of CD22 molecules. MATERIALS AND METHODS: Purified B cells and basophils were obtained from normal human volunteers by using a MACS magnetic cell sorting system and anti-CD19 and anti-Fc epsilon R1 antibodies, respectively. RT-PCR and sequencing of CD22 mRNA were performed in the exons 3 to 8. Western blotting analysis of CD22 was also performed. RESULTS: The sequence of CD22 mRNA extracted from the basophils was the same as that of B cells in exons 3 to 8 (epitopes recognized by Leu14, B3, and 4KB128 were translated from exons 4 and 5). Reduced CD22 peptide extracted from the basophils reacted to Leu14 as well as B3 and 4KB128, and the molecular size of the reduced and nonreduced products was 130 kDa as expected. CONCLUSION: Disulfide bonds and the resulting 3D conformation of the CD22 molecules may have important roles in the difference of antigenicity of CD22 beta in B cells (CD22 beta 1) and basophils (CD22 beta 2). The difference in molecular structure surrounding the ligand-binding domain of CD22 may imply a specialization of the conformational forms of CD22 according to the ligand isoforms. PMID- 11882358 TI - Translocation of Ku86/Ku70 to the multiple myeloma cell membrane: functional implications. AB - OBJECTIVE: Since the central hallmarks of human multiple myeloma (MM) are abnormalities in immunoglobulin (Ig) gene rearrangement, IgH class switching, and DNA damage repair, and since Ku86 and Ku70 proteins are central to these processes, aberrant Ku function may play a role in MM pathogenesis. Our prior studies demonstrated a 69-kDa Ku86 variant in freshly isolated patient MM cells that confers sensitivity to DNA damage. We also showed that Ku86 on the cell surface of CD40-activated MM cells mediates homotypic tumor cell adhesion, as well as heterotypic adhesion to bone marrow stromal cells. We here define the mechanism and functional significance of CD40-induced Ku translocation from the cytoplasm to the cell membrane in MM cells vs normal B cells. MATERIALS AND METHODS: We examined Ku86 and Ku70 translocation following CD40 activation in human MM cells vs normal tonsillar B lymphocytes. We then identified the functional sequelae of membrane Ku86 and Ku70 expression on CD40-activated human MM cells. RESULTS: CD40 activation induces translocation of both Ku86 and Ku70 to the cell surface of MM cells, but not normal tonsillar B cells. Moreover, CD40 activation triggers Ku association with CD40 only in CD40-activated MM cells. Finally, CD40-activated MM cells adhere to fibronectin and are protected against apoptosis triggered by irradiation or doxorubicin; conversely, antibodies to Ku both inhibit tumor cell binding and restore sensitivity to these agents. CONCLUSION: These results demonstrate functional significance of Ku translocation to the cell membrane of CD40-activated human MM cells. Therefore, targeting Ku86 and Ku70, with blocking peptides for example, might serve as a novel treatment strategy in human MM. PMID- 11882359 TI - Leukemic B cells clonally identical to myeloma plasma cells are myelomagenic in NOD/SCID mice. AB - OBJECTIVE: In multiple myeloma (MM), the immunoglobulin gene rearrangement characterizing malignant plasma cells is unique. For a patient with multiple myeloma who underwent a B-cell leukemic blast transformation, using the immunoglobulin molecular signature, we characterized the clonal relationship to autologous plasma cells and the impact on normal polyclonal B-lymphocyte populations. METHODS: Single-cell reverse transcriptase polymerase chain reaction (RT-PCR)/PCR was used to determine the clonal relationship between autologous MM plasma cells and leukemic B cells. A murine xenograft model was used to determine the myelomagenic potential of the leukemic B cells. RESULTS: Single-cell analysis showed that circulating leukemic-phase cells were clonotypic, with an IgH VDJ sequence identical to that of diagnosis plasma cells. Analysis of IgH transcripts indicates MM clonal dominance over normal B-cell components of the immune system at diagnosis and during leukemic disease. Leukemic B cells were xenografted to irradiated NOD/SCID mice, leading to lytic bone lesions and clonotypic cells in murine BM. Although human cells in murine BM expressed CD138, a marker largely absent from ex vivo leukemic cells, the expression of CD45, CD19, and CD20 confirmed that engrafting cells were mature, probably late-stage B cells rather than plasma cells. CONCLUSIONS: Leukemic B cells are able to exert strong clonal dominance over normal components of the immune system, colonize the murine BM in a xenograft model, and disrupt normal bone metabolism leading to lytic bone lesions. This supports the hypothesis that clonotypic MM B cells are reservoirs of disease that persist throughout therapy and give rise to relapse. PMID- 11882360 TI - Acquired uniparental disomy of chromosome 9p is a frequent stem cell defect in polycythemia vera. AB - OBJECTIVE: Clonal stem cell proliferation and increased erythrocyte mass are hallmarks of the myeloproliferative disorder polycythemia vera (PV). The molecular basis of PV is unknown. METHODS: We carried out a genome-wide screening for loss of heterozygosity (LOH) and analyzed candidate genes within the LOH loci. RESULTS: Three genomic regions were identified on chromosomes 9p, 10q, and 11q. The presence of these LOHs in both myeloid and lymphoid cells indicated their stem cell origin. The 9pLOH prevalence is approximately 33% and is the most frequent chromosomal lesion described in PV so far. We report that the 9pLOH is due to mitotic recombination and therefore remains undetectable by cytogenetic analysis. Nineteen candidate genes were selected within the 9pLOH region for sequencing and expression analysis. No mutations were found in these genes; however, unexpectedly, increased expression of the transcription factor NFI-B was detected in granulocytes and CD34(+) cells in PV with 9pLOH. Since a member of the NFI gene family (NFI-X) was reported to result in TGF-beta resistance when overexpressed in vitro (TGF-beta is a known inhibitor of hematopoiesis), we transfected the NFI-B gene to the mouse 32D cell line. We found that overexpression of the NFI-B gene confers TGF-beta resistance in vitro. CONCLUSIONS: We characterized a new region on chromosome 9p frequently involved in LOH in PV. Analysis of genes within this 9pLOH region revealed increased expression of the NFI-B gene. Our in vitro studies suggest that TGF-beta resistance may be the physiologic mechanism of clonal stem cell expansion in PV. PMID- 11882361 TI - The lymphoid protein tyrosine phosphatase Lyp interacts with the adaptor molecule Grb2 and functions as a negative regulator of T-cell activation. AB - OBJECTIVE: Following activation of T cells, phosphorylation of tyrosine residues occurs through a complex signaling process involving protein tyrosine kinases, phosphatases, and a variety of adapter molecules including Grb2. We have attempted to identify new signaling molecules that are important for the activation response. METHODS: Using a protein interaction screening protocol based on phage display, T-cell signaling components that associate with the adapter molecule, Grb2, the lymphoid-specific tyrosine phosphatase Lyp was identified. Using transcriptional reporter assays, the role of Lyp in T-cell activation was studied by overexpression of wild-type or catalytically inactive mutants of Lyp. RESULTS: A GST fusion containing the C-terminal SH3 domain of Grb2 bound to the nucleotide exchange factor Sos or Grb2-associated binder 2 (Gab2). In contrast, the N-terminal SH3-containing fusion bound to the protein tyrosine phosphatase Lyp. Grb2 was co-immunoprecipitated with Lyp in 293T cells overexpressing both proteins. Using Northern blot analysis, Lyp was found to be expressed predominantly in hematopoietic tissue, including spleen, lymph node, thymus, peripheral blood leukocytes, bone marrow, and fetal liver. Two human T cell lines, Jurkat and HuT78, expressed both Lyp mRNA and protein. Overexpression of wild-type Lyp or a catalytically inactive, substrate-trapping mutant (D195A) in Jurkat cells inhibited transcriptional activity initiated by anti-CD3 and anti CD28 antibodies. In contrast, two other catalytically inactive mutants (R233M or C227S) had no effect. CONCLUSION: These data demonstrate a novel interaction between the phosphatase Lyp and the adaptor Grb2 and are consistent with a negative regulatory role for Lyp in T-cell signaling. PMID- 11882362 TI - Ex vivo expansion, maturation, and activation of umbilical cord blood-derived T lymphocytes with IL-2, IL-12, anti-CD3, and IL-7. Potential for adoptive cellular immunotherapy post-umbilical cord blood transplantation. AB - OBJECTIVES: We investigated whether umbilical cord blood (UCB) T cells could be ex vivo expanded and activated in short-term culture for potential utilization as adoptive cellular immunotherapy post-umbilical cord blood transplantation (UCBT). METHODS: Fresh UCB mononuclear cells (MNCs) were isolated by Ficoll density centrifugation. Cryopreserved UCB mononuclear cells were thawed and washed with 2.5% human serum albumin and 5% dextrose in isotonic saline. The nonadherent MNC fraction were then plated in a serum-free cocktail of IL-2, IL-12, and anti-CD3 with and without IL-7 for 48 hours. Proliferation, cytotoxicity, TH1 (IFN-gamma), CD25, and CD45RO assays were performed. RESULTS: Proliferation studies demonstrated a significant increase in the proliferative ability of the UCB MNCs incubated in anti-CD3, IL-2, IL-12, and IL-7 (fresh--p < 0.005, and thawed--p < 0.001). The combination of all four agonists significantly induced expression of CD45 RO (fresh--p < 0.05, and thawed--p < 0.001) in both the CD4(+) and CD8(+) T cells expressing CD25 (fresh UCB--p < 0.01 [CD4] and p < 0.005 [CD8], respectively; thawed UCB--p < 0.001 [CD4] and p < 0.001 [CD8]). Intracellular cytokine profiles also revealed a significant increase in the production of IFN gamma (TH1 cells) (fresh UCB--p < 0.005, and thawed UCB--p < 0.001). The combination also significantly increased the killing of K562-labeled target cells (fresh--p < 0.0001, and thawed--0.731 +/- 0.03 vs 0.16 +/- 0.01) (p < 0.001). CONCLUSIONS: These data suggest that the ex vivo combination of IL-2, IL-12, anti CD3, and IL-7 significantly enhances the proliferation, activation, maturation, and cytotoxic potential of UCB T cells of both fresh and thawed UCB MNC. Further studies, however, are required to determine whether these ex vivo--expanded MNC could also potentially exacerbate acute or chronic graft-vs-host disease and/or other toxicities if utilized post-UCBT. PMID- 11882363 TI - Genetic analysis of patients with leukocyte adhesion deficiency: genomic sequencing reveals otherwise undetectable mutations. AB - OBJECTIVE: The aim of this study was to analyze mutations in DNA from patients with leukocyte adhesion deficiency (LAD), an immunodeficiency caused by absence of the beta(2) subunit (CD18) of the leukocyte integrins LFA-1 (CD11a/CD18), Mac 1 (CD11b/CD18), p150,95 (CD11c/CD18), and CR4 (CD11d/CD18). METHODS: We developed genomic DNA PCR sequencing to detect mutations not only in exons but also in introns. RESULTS: Eight LAD patients were analyzed, of which five had homozygous mutations, i.e., a 0.8-kb deletion, a branchpoint mutation in intron 5 causing mRNA missplicing, a nonsense mutation, and two missense mutations. Four of these mutations are novel. We cotransfected the two mutant CD18 proteins with normal CD11a, b, or c in COS cells. This resulted in absence of all three beta(2) integrins on the surface of cells transfected with CD18(252Arg). However, CD18(593Cys) supported some LFA-1 and p150,95 formation in COS cells. The other three patients were compound heterozygotes in which only one allele had previously been characterized, because the other alleles were undetectable at the cDNA level. We identified the unknown mutations as a novel two-nucleotide deletion, a nonsense mutation, and a single nucleotide deletion. CONCLUSION: Our method allows identification of mutations in CD18 from genomic DNA. This opens the possibility of early prenatal diagnosis of LAD and reliable carrier detection. PMID- 11882364 TI - Constitutive activation of STAT3 and STAT5 is induced by leukemic fusion proteins with protein tyrosine kinase activity and is sufficient for transformation of hematopoietic precursor cells. AB - OBJECTIVE: Signal transducers and activators of transcription (STAT) factors are critical mediators in the signal transduction of cytokine receptors. In hematopoietic and epithelial cells, activation of STAT 1 induces apoptosis and growth arrest, whereas activation of STAT3 and STAT5 transduces growth-promoting signals. We and others have previously described a high expression and constitutive activation of STAT1, 3, and 5 in AML blasts. In this report we focused on the mechanisms and also the biologic relevance of STAT activation in AML.Results. RESULTS: We report here that a constitutive STAT activation can be detected in up to 95% of primary AML blasts. In vitro, neither induction of the leukemic fusion protein PML-RAR alpha in U937 cells nor expression of transforming ras-mutants, but several leukemic protein tyrosine kinase (PTK), strongly induced activation of STAT3 and 5. Stable transfection of BA/F3 cells with TEL-JAK2, TEL-ABL, and BCR-ABL resulted in IL-3 independent growth and strong activation of STAT3 and STAT5 by TEL-JAK2 and TEL-ABL, but not by BCR-ABL. In addition, expression of constitutive active mutants of STAT3 and STAT5 alone were sufficient to transform BA/F3 cells. CONCLUSIONS: These results show that STAT3 and STAT5 are activated in the majority of primary AML blasts and are major targets of leukemic fusion proteins with PTK activity. However, the STAT activation pattern by leukemic PTKs differed significantly and might reflect their transforming potential in acute (TEL-JAK2 and TEL-ABL) and chronic leukemias (p210BCR-ABL). The transforming capacity of constitutively activated STAT3 and STAT5 mutants strongly supports their fundamental role in the process of malignant transformation in hematopoietic cells. PMID- 11882365 TI - Ceramide and sphingosine rapidly induce apoptosis of murine mast cells supported by interleukin-3 and stem cell factor. AB - OBJECTIVE: Ceramide and sphingosine, generated by sphingomyelinase-mediated hydrolysis of sphingomyelin, which packs tightly in the bilayer of the plasma membrane, have been proposed as intracellular mediators of apoptotic signals. However, precise function of endogenous sphingomyelin-cycle metabolites in mast cells has been unclear. Thus, we sought to define the involvement of ceramide and sphingosine in apoptotic pathways of mast cells. MATERIALS AND METHODS: We examined the effect of cell-permeable C(2)-ceramide, sphingosine, and sphingomyelinase on survival of murine bone marrow-derived cultured mast cells (BMCMC) supported by recombinant interleukin-3 (rIL-3) and/or recombinant stem cell factor (rSCF). Downstream signaling pathways of C(2)-ceramide and sphingosine were analyzed by using caspase inhibitors. RESULTS: C(2)-ceramide, sphingosine, and sphingomyelinase induced apoptosis in BMCMC in the presence of rIL-3 and/or rSCF, and Z-VAD-fmk (a broad caspase inhibitor), Z-DEVD-fmk (a caspase 3 inhibitor), and Z-IETD-fmk (a caspase 8 inhibitor) partially prevented apoptosis of BMCMC induced by C(2)-ceramide but not sphingosine. CONCLUSION: The present results suggest that ceramide and sphingosine may function as intracellular mediators of apoptotic signals in mast cells, which override survival signals from IL-3 and SCF. In addition, caspases may be partially involved in ceramide- but not sphingosine-mediated apoptosis of mast cells. PMID- 11882366 TI - Highly active antiretroviral therapy and allogeneic CD34(+) peripheral blood progenitor cells transplantation in an HIV/HCV coinfected patient with acute myeloid leukemia. AB - OBJECTIVE: To evaluate the safety, feasibility, and efficacy of allogeneic stem cell transplantation (SCT) for acute myelogenous leukemia (AML) in a young female coinfected by HIV and HCV undergoing highly active antiretroviral therapy (HAART). PATIENTS AND METHODS: A 33-year-old female HIV(+), HCV(+) in complete remission after standard chemotherapy was submitted to CD34(+) selected allogeneic transplantation from her HLA-identical HIV(-) brother after myeloablative regimen. HAART was started before transplantation, achieving a reduction of HIV load to undetectable levels. GVHD prophylaxis was carried out with cyclosporine A alone. RESULTS: The patient achieved prompt and durable engraftment with acute GVHD grade II easily managed with steroids; CMV prophylaxis was prolonged, no clinically relevant infectious complications developed early after transplantation and during follow-up. HIV viremia was controlled by HAART although medication adherence was reduced early after transplantation and required drug adjustment. There was a gradual recovery of immune cells with normal CD4-cell count 39 months after engraftment, a significantly higher level than before transplantation. At 39 months post transplantation follow-up the patient is alive and in continuous complete remission with undetectable levels of plasma HIV RNA on HAART. CONCLUSION: The introduction of HAART has recently changed the paradigm of AIDS, allowing the control of HIV replication, the reduction of opportunistic infections, and the overall improvement of survival. One may therefore reconsider the current exclusion of patients with AIDS and a concomitant lethal malignancy from programs of high-dose chemotherapy and stem cell transplantation, as suggested by this report. PMID- 11882368 TI - Reversal of neuronal polarity characterized by conversion of dendrites into axons in neonatal rat cortical neurons in vitro. AB - The mechanisms for the establishment and maintenance of cell polarity in neurons are not well understood. Axon regeneration from dendrites has been reported after axotomy near the cell body in vivo. We report here in vitro a reversal of neuronal polarity characterized by the conversion of dendrites into axons. We isolated neurons from the neonatal rat cerebral cortex. Neurons that exhibited an apical dendrite with a length of >100 microm were monitored for 3 days in culture. In 66% of neurons examined, a new axon, as identified by reactivity with an antibody to dephosphorylated tau or by lack of reactivity with an antibody to the a and b isoforms of microtubule-associated protein 2, appeared to form from the tip of the original dendrite. Further analysis of such neurons revealed that the distal half of the original dendrite became positive for dephosphorylated tau or negative for microtubule-associated protein 2. Time-lapse video microscopy demonstrated the conversion of the original dendrite into an axon without dendritic retraction. Axon regeneration from dendritic tips required a significantly longer time than axon regeneration from minor processes. Our observations thus demonstrate in vitro a time-consuming reversal of neuronal polarity and the conversion of a dendritic cytoskeleton into an axonal one. PMID- 11882367 TI - Astroglial in vivo response to cocaine in mouse dentate gyrus: a quantitative and qualitative analysis by confocal microscopy. AB - Astrocytes have been proved to play a critical role in neuromodulation, neuroprotection, pH maintenance, axon guidance control during development, homeostasis preservation and blood brain barrier maintenance in the CNS (Kimmelberg and Norenberg, 1989). Quantitative changes in the expression of glial fibrillary acidic protein (GFAP), a cytoskeletal intermediate filament protein exclusively expressed in astrocytes (Bignami et al, 1972), have been observed after administration of alcohol (Framke, 1995), morphine (Beitner-Johnson et al., 1993), amphetamine and its derivates (Aguirre et al., 1999), cannabinoids (Suarez et al., 2000), nicotine (Janson and Moller, 1993), caffeine (Marret et al., 1993) and prenatal exposure to cocaine (Clarke et al., 1996; Nassogne et al., 1998). However, the general astrocytic response to drugs of abuse is still far from being defined. In the present study we examined the in vivo astroglial response to cocaine in mouse dentate gyrus, the hippocampus being a common target of neurotoxic agents (Walsh and Emerich, 1988) which has a prominent effect on learning and memory processes (Eichenbaum et al., 1992). Quantitative changes in immunoreactivity of GFAP were investigated 24 h after acute and repeated daily administration of intraperitoneal cocaine (20 mg/kg). Drug-induced morphological alterations and spatial distribution of astrocytes were evaluated by means of confocal microscope. The results show that, compared to control animals, GFAP expression is two-fold enhanced after a single cocaine injection, still significantly higher after seven consecutive daily administrations, but not statistically different after prolonged (14 days) drug treatment. Moreover, morphological and morphometric analyses reveal significant modifications in astrocytic numbers, cell size and shape complexity. These data demonstrate that in mouse dentate gyrus, cocaine exposure differently affects the expression of GFAP and induces strong changes in astrocytes proliferation rate and cell morphology. Taken together, our findings provide the first in vivo quantitative and qualitative evaluation of astrocytic response to several regimens of cocaine in adult animals brain. PMID- 11882369 TI - Thyroid hormone regulates neurotransmitter release in neonatal rat hippocampus. AB - Thyroid hormone is essential for the normal maturation and function of the mammalian CNS. Thyroid hormone deficiency during a critical period of development profoundly affects cognitive functions such as learning and memory. However, the possible electrophysiological alterations that could underlie these learning deficits in hypothyroid animals remain largely unexplored. In this work, we have studied the possible effect of thyroid hormone on short-term synaptic plasticity, which is hypothesized to be a neural substrate of short-term memory. We compared short-term modification of the excitatory postsynaptic potential in hippocampal slices between control and hypothyroid rats. Electrophysiological studies reveal that paired-pulse facilitation is strongly altered in the hypothyroid rats. In addition, hypothyroid rats exhibit an increase in the Ca(2+)-dependent neurotransmitter release. These alterations are basically reversible when thyroid hormone is administered. In order to examine the possible molecular mechanisms underlying these synaptic changes, we compared the expression of synapsin I, synaptotagmin I, syntaxin, and alpha-Ca(2+)/calmodulin kinase II between control and hypothyroid hippocampus. Our results show that the levels of synapsin I and synaptotagmin I are increased in the hypothyroid rats, which suggests that the genes encoding these proteins are implicated in the action of thyroid hormone on neurotransmitter release. Taken together, the results from this study suggest that thyroid hormone may modulate the probability of neurotransmitter release. PMID- 11882370 TI - Alpha3 integrin receptors contribute to the consolidation of long-term potentiation. AB - Several lines of evidence suggest that integrin receptors play a pivotal role in consolidation of long-term potentiation (LTP), but which of the many integrin dimers are involved remains to be discovered. The present study used an LTP reversal paradigm to test if alpha3 integrins make an important contribution. Function blocking alpha3 monoclonal antibodies or vehicle were locally infused into recording sites in field CA1 of rat hippocampal slices and LTP induced with theta burst stimulation. Low frequency trains of pulses were applied 30 min after the theta bursts. Previous work indicates that low frequency stimulation reverses LTP when applied immediately after induction but is largely ineffective after 30 45-min delays. If the antibodies were to block consolidation, then they should extend the period over which potentiation is vulnerable to disruption. There was no detectable difference between the two groups in the initial degree of LTP or within slice decay of potentiation 1-10 min after induction; a small but reliable decay occurred from 10 to 30 min with antibody treatment (P<0.01) but not in control slices. Percent potentiation was not statistically different for vehicle (55 +/- 19%, mean +/- S.D.) and anti-alpha3 (43 +/- 21%) slices at 30 min post theta bursts. Five-Hz stimulation ("theta pulse" stimulation) 30 min after induction caused a reduction of LTP. The percent loss of potentiation after the 1 min trains was greater in the antibody-treated slices than in controls (98 +/- 4% vs. 62 +/- 28%, P<0.01, U-test) and correlated (r=0.84, alpha3 slices) with the percent LTP present prior to low frequency stimulation, as expected if the stimulation reversed potentiation. Recovery occurred in both groups but percent LTP was significantly smaller in experimental slices at 10 min post-theta pulses (5 +/- 11% vs. 36 +/- 15%, P<0.01). Recovery continued for 20 min after theta pulses and, in accordance with earlier work, was nearly complete for the control slices (50 +/- 19% vs 55 +/- 15%, 40 min post- vs. immediately pre-theta pulses). LTP remained depressed after 40 min of recovery in the anti-alpha3 slices (23 +/- 19% vs. 43 +/- 21%) at which point it was substantially less than that found in controls (P<0.01). Western blots with anti-alpha3 antibodies identified a polypeptide with the molecular mass (155 kDa) expected for the alpha3 subunit and further showed that it is broadly distributed in brain. Subcellular fractionation experiments demonstrated that alpha3 is concentrated in synaptic membranes over homogenates to about the same degree as the GluR1 subunit of the alpha-amino-3 hydroxy-5-methyl-4-isoxazole propionate-type glutamate receptor. From these results we suggest that alpha3-containing integrins are localized to synapses and are needed to stabilize a slowly decaying form of LTP. The findings also show that vulnerability to reversal can be used in place of extended recording sessions in studying consolidation. PMID- 11882371 TI - Altered forebrain neurotransmitter responses to immobilization stress following 3,4-methylenedioxymethamphetamine. AB - (+/-)3,4-Methylenedioxymethamphetamine (MDMA, "ecstasy") is an increasingly popular drug of abuse that acts as a neurotoxin to forebrain serotonin neurons. The neurochemical effects of the serotonin depletion following high doses of MDMA were investigated in response to acute immobilization stress. Male rats were treated with a neurotoxic dosing regimen of MDMA (10 mg/kg, i.p. every 2 h for four injections) or equivalent doses of saline. Seven days after treatment, in vivo microdialysis was used to assess extracellular dopamine and serotonin in the dorsal hippocampus and prefrontal cortex during 1 h of immobilization stress. In saline treated control rats, serotonin in the hippocampus and serotonin and dopamine in the prefrontal cortex were increased during immobilization stress. Rats pretreated with MDMA, however, showed blunted neurotransmitter responses in the hippocampus and the prefrontal cortex. In the drug pretreated rats, basal serotonin levels in the hippocampus, but not the prefrontal cortex, were lower compared to saline pretreated controls. Stress-induced increases in plasma corticosterone and body temperature were not affected by the pretreatment condition. From these studies we suggest that depletion of serotonin stores in terminal regions with the neurotoxin MDMA compromises the ability of the serotonergic neurons to activate central systems that respond to stressful stimuli. This altered responsiveness may have implications for long-term functional consequences of MDMA abuse as well as the interactions between the serotonergic system and stress. PMID- 11882372 TI - Involvement of inducible nitric oxide synthase in inflammation-induced dopaminergic neurodegeneration. AB - The loss of dopaminergic neurones in the substantia nigra with Parkinson's disease may result from inflammation-induced proliferation of microglia and reactive macrophages expressing inducible nitric oxide synthase (iNOS). We have investigated the effects of the supranigral administration of lipopolysaccharide on iNOS-immunoreactivity, 3-nitrotyrosine formation and tyrosine hydroxylase immunoreactive neuronal number, and retrogradely labelled fluorogold-positive neurones in the ventral mesencephalon in male Wistar rats. Following supranigral lipopolysaccharide injection, 16-18 h previously, there was intense expression of NADPH-diaphorase and iNOS-immunoreactivity in non-neuronal, macrophage-like cells. This was accompanied by intense expression of glial fibrillary acidic protein-immunoreactive astrocytosis in the substantia nigra. There were also significant reductions in the number of tyrosine hydroxylase(50-60%)- and fluorogold (65-75%)-positive neurones in the substantia nigra. In contrast, tyrosine hydroxylase-immunoreactivity in the ventral tegmental area was not altered. Pre-treatment of animals with the iNOS inhibitor, S-methylisothiourea (10 mg kg(-1), i.p.), led to a significant reduction of lipopolysaccharide induced cell death. Similar reduction of tyrosine hydroxylase-immunoreactivity and fluorogold-labelled neurones in the substantia nigra following lipopolysaccharide administration suggests dopaminergic cell death rather than down-regulation of tyrosine hydroxylase. We conclude that the expression of iNOS- and 3-nitrotyrosine-immunoreactivity and reduction of cell death by S methylisothiourea suggest the effects of lipopolysaccharide may be nitric oxide mediated, although other actions of lipopolysaccharide (independent of iNOS induction) cannot be ruled out. PMID- 11882373 TI - Phase- and position-disparity coding in the posteromedial lateral suprasylvian area of the cat. AB - The posteromedial lateral suprasylvian area of the cat is known to be involved in the analysis of motion and motion in depth. However, it remains unclear whether binocular cells in the posteromedial lateral suprasylvian area rely upon phase or positional offsets between their receptive fields in order to code binocular disparity. The present study aims at clarifying more precisely the neural mechanisms underlying stereoperception with two objectives in mind. First, to determine whether cells in the posteromedial lateral suprasylvian area code phase disparities. Secondly, to examine whether the cells sensitive to phase disparity are the same as those which code for position disparities or whether each group represent a different sub-population of disparity-sensitive neurons. We investigated this by testing both types of disparities on single neurons in this area. The results show that the vast majority of cells (74%), in the posteromedial lateral suprasylvian area, are sensitive to relative interocular phase disparities. These cells showed mostly facilitation (95%) and a few (5%) summation interactions. Moreover, most cells (81%) were sensitive to both position and phase disparities. The results of this study show that most binocular cells in the posteromedial lateral suprasylvian area are sensitive to both positional and phase offsets which demonstrate the importance of this area in stereopsis. PMID- 11882374 TI - CNS inputs to the suprachiasmatic nucleus of the rat. AB - The neural circuits that modulate the suprachiasmatic nucleus (SCN) of the rat were studied with the retrograde transneuronal tracer--pseudorabies virus. First order afferents were also identified using cholera toxin beta subunit. Olfactory processing regions (viz., main olfactory bulb, anterior olfactory nucleus, taenia tecta, endopiriform nucleus, medial amygdaloid nucleus, piriform cortex, and posteriomedial cortical amygdaloid nucleus) were virally labeled. The subfornical organ directly innervates SCN; two other circumventricular organs: organum vasculosum of the lamina terminalis and area postrema provide multisynaptic inputs. Direct limbic afferents arise from lateral septum, bed nucleus of the stria terminalis, amygdalohippocampal zone, and ventral subiculum; multineuronal connections come from the basolateral and basomedial amygdaloid nuclei, ventral hippocampus, amygdalopiriform area, as well as lateral entorhinal, perirhinal, and ectorhinal cortices. Most preoptic regions project directly to SCN. Multisynaptic inputs come from the lateral preoptic region. Hypothalamic inputs originate from the anterior, arcuate, dorsal, dorsomedial, lateral, paraventricular, posterior, periventricular posterior, retrochiasmatic, subparaventricular, ventromedial and tuberomammillary nuclei. Paraventricular thalamic nucleus, intergeniculate leaflet and zona incerta directly innervate SCN. Polyneuronal inputs arise from the subparafascicular parvicellular thalamic nucleus. Brainstem afferents originate from the pretectum, superior colliculus, periaqueductal gray matter, parabrachial nucleus, pedunculopontine nucleus, raphe system, locus coeruleus, nucleus incertus and reticular formation. Nucleus tractus solitarius, C3 catecholamine region, rostral ventrolateral medulla and spinal trigeminal nucleus provide indirect inputs. We propose that the SCN receives feedback primarily from interoceptive systems such as the circumventricular, autonomic, and neuroendocrine systems that are important in the central regulation of glucose metabolism (e.g., insulin and glucocorticoids). PMID- 11882375 TI - Characterisation of rat superficial superior colliculus neurones: firing properties and sensitivity to GABA. AB - Physiological, pharmacological and morphological properties of superficial superior colliculus neurones (n=93) were characterised using whole-cell patch clamp recordings in rat brain slices. Six cell types (narrow- and wide-field vertical, horizontal, piriform, marginal and stellate) were identified based on Lucifer Yellow labelling but no cell type-specific spike pattern could be identified. Resting membrane potentials were homogeneous (mean: -67.1 +/- 0.7 mV, n=48), and spike frequencies ranged from 10 to 70 Hz (80 pA current injection). About 66% of the cells displayed regular and sustained spike production, throughout all neuronal categories. Rebound spikes and spontaneous activity were observed frequently in all cell types. Synaptically evoked action potentials appeared as single spikes (mean amplitude: 76.0 +/- 3.2 mV, n=34) followed by a fast after-hyperpolarising potential (mean amplitude: 25.4 +/- 1.4 mV, n=34) and variable late potentials (late after-depolarising and/or -hyperpolarising). Pharmacologically, a characterisation using GABA and its subtype-specific agonists indicated a strong inhibitory influence of this transmitter system on >90% of cells. The GABA(A) receptor agonist, 4,5,6,7-tetrahydroisoxazolo[5,4 c]pyridin-3-ol (100 microM), caused a reversible hyperpolarisation (approximately 9 mV) and spike inhibition of all neurones studied. This was more pronounced for intrinsic than for synaptically evoked spikes. Assessment of the GABA(C) receptor agonist, cis-4-aminocrotonic acid (1 mM), also revealed a hyperpolarisation (approximately 3 mV) and an inhibitory action on firing, but this was not as potent and homogeneous, compared to the GABA(A) receptor agonist. Further, the GABA(B) receptor agonist, baclofen (50-100 microM), had more variable (hyperpolarising, depolarising or no change) effects on the membrane potential. It showed little modulation of current-induced action potentials but fully blocked synaptic spikes. Assessment of GABA receptor antagonist actions revealed the presence of weak tonic and strong phasic GABA(A) receptor-mediated inhibition in the superficial superior colliculus: application of the GABA(A) receptor antagonist, bicuculline (100 microM), led to a generally enhanced excitability and depolarisation (approximately 5 mV). Intrinsic firing was somewhat enhanced, but synaptic spiking was drastically potentiated and prolonged. In contrast, 1,2,5,6-tetrahydro-(pyridin-4-yl) methylphosphinic acid (TPMPA; 100 microM), the GABA(C) receptor antagonist, produced little effect on these physiological parameters. The GABA(B) receptor antagonist, CGP35348 (200 microM), caused a partial inhibition of late after-hyperpolarising potentials (approximately 30%). Uptake of GABA contributes little to endogenous inhibition in the superior colliculus slice preparation, as suggested by the action of GABA uptake inhibitors SKF89976 (50-100 microM) and nipecotic acid (200-500 microM), both had no obvious effect on physiological parameters. However, in the presence of these compounds, sub-maximal inhibitory actions of GABA were potentiated. In conclusion, different cell types in the superficial superior colliculus do not display distinct physiological properties and are subject to strong inhibitory modulation. We therefore suggest that signal processing in this brain region does not require cell type-specific encoding of information. In line with evidence provided by previous in vivo investigations, identification of visual stimuli and orientation responses appears to be realised via the network properties of the receptive fields that form topographic maps. PMID- 11882376 TI - Cerebellar input to magnocellular neurons in the red nucleus of the mouse: synaptic analysis in horizontal brain slices incorporating cerebello-rubral pathways. AB - We studied the synaptic input from the nucleus interpositus of the cerebellum to the magnocellular division of the red nucleus (RNm) in the mouse using combined electrophysiological and neuroanatomical methods. Whole-cell patch-clamp recordings were made from brain slices (125-150 microm) cut in a horizontal plane oriented to pass through both red nucleus and nucleus interpositus. Large cells that were visually selected and patched were injected with Lucifer Yellow and identified as RNm neurons. Using anterograde tracing from nucleus interpositus in vitro, we examined the course of interposito-rubral axons which are dispersed in the superior cerebellar peduncle. In vitro monosynaptic responses in RNm were elicited by an electrode array placed contralaterally in this pathway but near the midline. Mixed excitatory post-synaptic potentials (EPSPs)/inhibitory post synaptic potentials (IPSPs) were observed in 48 RNm neurons. Excitatory components of the evoked potentials were studied after blocking inhibitory components with picrotoxin (100 microM) and strychnine (5 microM). All RNm neurons examined continued to show monosynaptic EPSPs after non-N-methyl-D aspartate (NMDA) glutamate receptor components were blocked with 10 microM 6,7 dinitroquinoxaline-2,3-dione or 5 microM 2,3-dihydro-6-nitro-7-sulfamoyl-benzo(f) quinoxaline (NBQX; n=12). The residual potentials were identified as NMDA receptor components since they (i) were blocked by the addition of the NMDA receptor antagonist, D,L-2-amino-5-phosphonovaleric acid (APV), (ii) were voltage dependent, and (iii) were enhanced by Mg(2+) removal. Inhibitory components of the evoked potentials were studied after blocking excitatory components with NBQX and APV. Under these conditions, all RNm neurons studied continued to show IPSPs. Blockade of GABA(A) receptors reduced but did not eliminate the IPSPs. These were eliminated when GABA(A) receptor blockade was combined with strychnine to eliminate glycine components of the IPSPs. Thus, IPSPs evoked by midline stimulation of the superior cerebellar peduncle, while blocking alpha-amino-3 hydroxy-5-methylisoxazole-4-propionic acid (AMPA) and NMDA receptors, raise the possibility of direct inhibitory inputs to RNm from the cerebellum. In summary we propose that the special properties of the NMDA receptor components are considered important for the generation of RNm motor commands: their slow time course will contribute a steady driving force for sustained discharge and their voltage dependency will facilitate abrupt transitions from a resting state of quiescence to an active state of intense motor command generation. PMID- 11882377 TI - ATP stimulates peripheral axons in human, rat and mouse--differential involvement of A(2B) adenosine and P2X purinergic receptors. AB - Receptors for ATP have been reported on peripheral nerve terminals. It is a widespread assumption that the axonal membrane does not possess this kind of chemosensitivity, although P2X purinoceptors have been found in isolated rat vagus nerve. Therefore, in the present study, effects of ATP and analogues were tested on the excitability of unmyelinated axons in isolated rat sural nerve, mouse dorsal roots, and human sural nerve. Bath application of ATP to all three types of axonal preparations increased axonal excitability, but the underlying receptors appear to differ in the various preparations. In rat sural nerve, alpha,beta-adenosine-5'-methylene triphosphate produced the strongest excitation. This effect was blocked by pyridoxal-phosphate-6-azophenyl-2',5'-disulphonic acid and indicates the presence of P2X receptors. In mouse dorsal roots, differences were found between fast and slow C-fibres. The latter responded to both P2X receptor and adenosine receptor agonists. In contrast, effects of ATP on faster conducting C-fibres seem to be caused exclusively by effects of ATP on adenosine receptors. Application of ATP also excited C-fibres in fascicles of isolated human nerve. The pharmacological profile indicates activation of A(2B) adenosine receptors. However, we could not detect P2X receptors in this preparation with our techniques. These data show that the ATP sensitivity of sensory neurones is not restricted to their terminals. Activation of axonal purinergic receptors may contribute to the transduction of sensory, including nociceptive, stimuli. PMID- 11882378 TI - Localization of functional calcitonin gene-related peptide binding sites in a subpopulation of cultured dorsal root ganglion neurons. AB - In this study we investigated whether cultured dorsal root ganglion (DRG) neurons from the adult rat express binding sites for calcitonin gene-related peptide (CGRP). These were identified on fixed cells by using CGRP labeled at the N terminal site with 1.4-nm gold particles. After 1 day in culture, about 20% of small to medium-sized DRG neurons showed CGRP-gold binding. Binding of CGRP-gold was dose-dependently reduced by coadministration of CGRP. The calcium imaging technique in living cells revealed that the bath administration of CGRP evoked an increase of the intracellular calcium in up to 30% of the DRG neurons tested. Both depletion of intracellular calcium stores by thapsigargin or using a calcium free medium blocked the CGRP-mediated increase of cytosolic calcium in most neurons. Thus intracellular and extracellular sources of calcium are relevant for the CGRP response. Using the whole-cell patch-clamp technique, about 30% of the neurons were found to exhibit an inward current and a depolarization upon administration of CGRP close to the neurons. Immunocytochemical double-labeling techniques showed that most of the CGRP-gold binding sites were expressed in unmyelinated (neurofilament 200-negative) DRG neurons. Most of the neurons with CGRP-gold binding sites also expressed the tyrosine kinase A receptor, and all of them showed CGRP-like immunoreactivity. This study shows, therefore, that a subpopulation of unmyelinated, peptidergic primary afferent neurons express CGRP binding sites that can be activated by CGRP in an excitatory direction. The binding sites may serve as autoreceptors because all of these neurons also synthesize CGRP. The activation of CGRP binding sites may sensitize primary afferent neurons and influence the release of mediators. PMID- 11882379 TI - Decreased calcium influx into the neonatal rat motor nerve terminals can recruit additional neuromuscular junctions during the synapse elimination period. AB - Individual skeletal muscle fibers in newborn vertebrates are innervated at a single endplate by several motor axons. During the first postnatal weeks, the polyneuronal innervation decreases in an activity-dependent process of synaptic elimination by axonal competition. Because synaptic activity depends strongly on the influx of calcium from the external media via presynaptic voltage-dependent calcium channels, we investigate the relationship between calcium channels, synaptic activity and developmental axonal elimination. We studied how several calcium channel blockers affect (after 1 h of incubation) the total number of functional axons per muscle fiber (poly-innervation index) of the Levator auris longus muscle of 6-day-old rats. We determined the poly-innervation index by gradually raising the stimulus amplitude and recorded the recruitment of one or more axons that produced a stepwise increment of the endplate potential.The L type channel blocker nitrendipine (1 microM) increased the mean poly-innervation index (35.79% +/- 3.91; P<0.05). This effect was not washed out with normal Ringer, although the poly-innervation index returned to the control value when high-calcium Ringer (5 mM) was used. The P-type channel blocker omega-agatoxin IVA (100 nM) also increased the number of recruitable endplate potentials (27.49% +/- 1.78; P<0.05), whereas N-type channel blocker omega-conotoxin-GVIA (1 microM) was ineffective (P>0.05). However, neither nitrendipine nor omega-agatoxin-IVA modified the poly-innervation index on high-calcium Ringer (P>0.05 in both cases). A more intense inhibition of calcium influx (by the sequential use of two calcium channel blockers) did not recruit any additional silent synapses. Moderately increasing the magnesium ions (by 500 microM) in the physiological solution produces a synaptic recruitment (36.78% +/- 2.1; P<0.05) similar to that with L- and P-type calcium channel blockers incubation. This magnesium effect was not washed with normal Ringer but a Ringer that is high in calcium can reverse it. The recruited endings were identified by selective activity-dependent loading with styryl dyes. Rhodaminated alpha-bungarotoxin-labeled acetylcholine receptors were present in the postsynaptic counterpart. Based on these findings we suggest that, before their complete retraction, functionally silent nerve terminals can be manifested or recovered if calcium influx is reduced by a calcium channel blocker or if external magnesium is increased. The normal activation of this calcium-dependent silencing mechanism during development may be related to the final loss of the supernumerary axons. PMID- 11882380 TI - Modulation of nerve growth factor in peripheral organs by estrogen and progesterone. AB - Nerve growth factor (NGF) synthesized in peripheral organs plays a critical role in the development and maintenance of the nervous system and also participates in processing nociceptive stimuli. Previous studies suggest that reproductive hormones may regulate the expression of NGF. Ovariectomies were performed on female mice, and mice were killed 24 h after hormone replacement to evaluate the effects of estrogen and progesterone on NGF in peripheral organs, specifically the uterus, bladder, heart, and salivary gland. Sham-operated intact mice and untreated ovariectomized mice served as controls. Immunohistochemistry demonstrated the presence of NGF, estrogen receptor-alpha, estrogen receptor beta, and progesterone receptors in these organs. Ovariectomy caused a significant decrease in NGF protein content in the uterus, and short term treatment of ovariectomized mice with estrogen and/or progesterone increased uterine NGF mRNA and restored NGF protein to concentrations similar to intact control mice. Ovariectomy did not affect NGF protein concentrations in the salivary gland, but treatment of ovariectomized mice with estrogen alone or in conjunction with progesterone stimulated concentrations of NGF protein that exceeded those observed in intact control or ovariectomized, untreated mice. NGF mRNA was increased in salivary glands from ovariectomized mice treated with progesterone alone or in combination with estrogen relative to other groups. NGF protein content of the hearts of ovariectomized mice treated with estrogen alone or in conjunction with progesterone was increased relative to intact controls and ovariectomized, untreated mice, but neither ovariectomy or hormone replacement affected NGF mRNA content in the heart. NGF protein content of the bladder was unaffected by ovariectomy or hormone treatment, and bladder NGF mRNA was unaffected by ovariectomy or hormone treatment. Collectively, these results indicate that reproductive hormones have the capacity to regulate NGF message and protein in a manner that varies among organs. Fluctuations in the expression of NGF, in conjunction with other factors, may help to explain gender differences in pain sensation and inflammatory response. PMID- 11882381 TI - Serotonin antagonizes the human neuronal alpha7 nicotinic acetylcholine receptor and becomes an agonist after L248T alpha7 mutation. AB - The effects of serotonin (5-hydroxytryptamine or 5HT) on chick alpha7 nicotinic receptors have already been described. However similar studies on human alpha7 receptors have been lacking. To begin to fill this deficiency, studies were made on wild-type and mutant human alpha7 (halpha7) receptors expressed in Xenopus oocytes or human BOSC 23 cells. In oocytes wild-type halpha7 receptors were blocked by 5HT, and this block was voltage-dependent. In contrast, 5HT acted as an agonist on halpha7-mutant receptors (L248T). Outside-out membrane-patches from BOSC 23 cells expressing halpha7-mutant receptors exhibited spontaneous channel openings of two conductance levels (59 pS and 76 pS) and short mean open time (0.9 ms). halpha7-Mutant channels activated by nicotine or 5HT displayed similar conductances and high Ca(2+) permeability; but longer duration (2.7 ms) than the spontaneous openings. Mutations at Cys190 and Cys191, in the extracellular N terminus of the human alpha7 gene, did not prevent receptor expression and incorporation in the oocyte membrane (determined by alpha-bungarotoxin binding). However, both 5HT and nicotine were incapable of gating the channels, indicating that the mutated Cys residues are in, or near, the 5HT- and nicotine-binding site. This is the first report that alpha7 receptors have spontaneous openings; and that 5HT is an agonist of halpha7-mutant receptors, and an antagonist of halpha7-wild-type receptors, through interactions at, or near the acetylcholine binding sites. PMID- 11882382 TI - FMRFamide modulates potassium currents in circadian pacemaker neurons of Bulla gouldiana. AB - The peptide FMRFamide (Phe-Met-Arg-Phe-NH(2)) is known to modulate the circadian pacemaker found in the eye of the marine snail Bulla gouldiana. In the present study, we investigated the cellular mechanisms underlying this modulation by examining the effects of FMRFamide on the membrane properties of the circadian pacemaker cells, known as basal retinal neurons in this preparation. Bath application of FMRFamide (0.1-1 microM) increased the membrane conductance, and hyperpolarized the membrane potential of these neurons. Next, perforated-patch recordings were used to demonstrate that FMRFamide reversibly increased the outward current amplitude due to an augmentation of a non-inactivating calcium independent current. Reversal potential of the tail currents and its dependence on extracellular potassium concentration suggested potassium ions as the charge carrier for this current. The peptide-modulated outward current was blocked by 54% after bath application of the potassium channel blocker tetraethylammonium chloride and completely blocked by substituting cesium for intracellular potassium. Voltage dependence, activation kinetics and tail current kinetics of the FMRFamide-modulated current were consistent with values found for the delayed rectifier current.Overall, our data suggest that FMRFamide modulates a delayed rectifier potassium current and at least one other, less voltage-dependent conductance. This provides a mechanistic explanation for FMRFamide's ability to both shift the phase and attenuate light-induced phase shifts of the circadian pacemaker in B. gouldiana. PMID- 11882383 TI - The protein kinase B/Akt signalling pathway in human malignancy. AB - Protein kinase B or Akt (PKB/Akt) is a serine/threonine kinase, which in mammals comprises three highly homologous members known as PKBalpha (Akt1), PKBbeta (Akt2), and PKBgamma (Akt3). PKB/Akt is activated in cells exposed to diverse stimuli such as hormones, growth factors, and extracellular matrix components. The activation mechanism remains to be fully characterised but occurs downstream of phosphoinositide 3-kinase (PI-3K). PI-3K generates phosphatidylinositol-3,4,5 trisphosphate (PIP(3)), a lipid second messenger essential for the translocation of PKB/Akt to the plasma membrane where it is phosphorylated and activated by phosphoinositide-dependent kinase-1 (PDK-1) and possibly other kinases. PKB/Akt phosphorylates and regulates the function of many cellular proteins involved in processes that include metabolism, apoptosis, and proliferation. Recent evidence indicates that PKB/Akt is frequently constitutively active in many types of human cancer. Constitutive PKB/Akt activation can occur due to amplification of PKB/Akt genes or as a result of mutations in components of the signalling pathway that activates PKB/Akt. Although the mechanisms have not yet been fully characterised, constitutive PKB/Akt signalling is believed to promote proliferation and increased cell survival and thereby contributing to cancer progression. This review surveys recent developments in understanding the mechanisms and consequences of PKB/Akt activation in human malignancy. PMID- 11882384 TI - G protein-coupled receptor signalling and cross-talk: achieving rapidity and specificity. AB - Activation of a given type of G protein-coupled receptor (GPCR) triggers a limited set of signalling events in a very rapid and specific manner. The classical paradigm of GPCR signalling was rather linear and sequential. Emerging evidence, however, has revealed that this is only a part of the complex signalling mediated by GPCR. Propagation of GPCR signalling involves cross regulation of many but specific pathways, including cross-talks between different GPCRs as well as with other signalling pathways. Moreover, it is increasingly apparent that GPCRs can activate both heterotrimeric G protein-dependent and G protein-independent signalling pathways. In this review, we discuss how the signallings initiated by GPCRs achieve rapidity as well as specificity, and how the GPCRs can cross-regulate other specific signalling pathways at the same time. New concepts regarding GPCR signalling have been arising to address this issue, which include multiprotein signalling complex and signalling compartment in microdomain concepts that enable close colocalization or even contact among the proteins engaged in the specific signal transduction. The final outcome of a stimulation of GPCR will thus be the sum of its own specific set of intracellular signalling pathways it regulates. PMID- 11882385 TI - G protein specificity: traffic direction required. AB - This review focuses on the coupling specificity of the Galpha and Gbetagamma subunits of pertussis toxin (PTX)-sensitive G(i/o) proteins that mediate diverse signaling pathways, including regulation of ion channels and other effectors. Several lines of evidence indicate that specific combinations of G protein alpha, beta and gamma subunits are required for different receptors or receptor-effector networks, and that a higher degree of specificity for Galpha and Gbetagamma is observed in intact systems than reported in vitro. The structural determinants of receptor-G protein specificity remain incompletely understood, and involve receptor-G protein interaction domains, and perhaps other scaffolding processes. By identifying G protein specificity for individual receptor signaling pathways, ligands targeted to disrupt individual pathways of a given receptor could be developed. PMID- 11882386 TI - Activation and inactivation of signal transducers and activators of transcription by ciliary neurotrophic factor in neuroblastoma cells. AB - Neurons in vivo are exposed to a variety of different growth factors and cytokines. A principal signalling pathway for ciliary neurotrophic factor (CNTF) like cytokines is the Janus kinase (Jak)/signal transducer and activator of transcription (STAT) system of kinases and transcription factors. In the human cell line (SH-SY5Y), STAT1 and STAT3 activation by CNTF-like cytokines showed tyrosine phosphorylation peaking at 0.5 h and inactivating within 2 h. Tyrosine phosphorylation of the receptor-associated tyrosine kinases Jak1 and Jak2 showed a similar time course of activation and inactivation in response to CNTF. The STAT1 response to the non-CNTF-like cytokine, interferon-gamma (IFN-gamma) did not inactivate. Inactivation to CNTF was not due to a decrease in CNTF receptor subunit gp130 or in levels of Jak1 or Jak2. STAT inactivation was inhibited by the protein kinase blocker H7 and a tyrosine phosphatase blocker, but not by inhibitors of protein kinase C, mitogen-activated protein kinase (MAPK) kinase, mTOR-P70/S6 kinase or phosphatidyl inositol-3-kinase (PI-3 kinase). Surprisingly, CNTF caused only a minor increase in levels of suppressors of cytokine signalling, SOCS-1 and SOCS-3. CNTF pretreatment desensitized the cells to the CNTF-like cytokines, leukemia inhibitory factor and oncostatin-M but not to IFN gamma. These results reveal a complex level of regulation of shared signalling pathways for cytokines that is dependent on both the type of cell and cytokine. PMID- 11882387 TI - Nongenomic action of progesterone in rat aorta: role of nitric oxide and prostaglandins. AB - The mechanism of action of progesterone (Pg) on rat vascular tissue was investigated. We obtained evidence that 10-nM Pg inhibited platelet aggregation at 1-5 min. Previously, we reported that nitric oxide (NO) mediated this antiaggregatory effect. Rat aortic strips (RAS) NO synthase (NOS) activity in response to "in vitro" treatment with other sex steroids hormones was measured. The stimulatory action of Pg on NO production was specific for ovarian hormones and depends on sex. The effect was nongenomic since cycloheximide did not suppress the increment in NO induced by Pg. Finally, we demonstrated that Pg (5 min) increased prostacyclin release (42-182% above control) in a dose-dependent manner (1-100 nM). Indeed, indomethacin (10 microM) completely suppressed the increment in citrulline levels induced by the hormone. These results suggest that Pg exerts a direct nongenomic action on rat aortic metabolism, which involves NOS and cyclooxygenase (COX) activation and a cross-talk between NO- and prostacyclin (PGI(2))-dependent pathways. PMID- 11882388 TI - Oxidative stress-induced calcium signalling in Aspergillus nidulans. AB - The effects of oxidative stress on levels of calcium ion (Ca(2+)) in Aspergillus nidulans were measured using strains expressing aequorin in the cytoplasm (Aeq(cyt)) and mitochondria (Aeq(mt)). When oxidative stress was induced by exposure to 10-mM H(2)O(2), the mitochondrial calcium response (Ca(mt)(2+)) was greater than the change in cytoplasmic calcium (Ca(c)(2+)). The Ca(mt)(2+) response to H(2)O(2) was dose dependent, while the increase in [Ca(c)(2+)] did not change with increasing H(2)O(2). The increase in both [Ca(c)(2+)] and [Ca(mt)(2+)] in response to oxidative stress was enhanced by exposure of cells to Ca(2+). The presence of chelator in the external medium only partially inhibited the Ca(mt)(2+) and Ca(c)(2+) responses to oxidative stress. Reagents that alter calcium fluxes had varied effects on the Ca(mt)(2+) response to peroxide. Ruthenium red blocked the increase in [Ca(mt)(2+)], while neomycin caused an even greater increase in [Ca(mt)(2+)]. Treatment with ruthenium red and neomycin had no effect on the Ca(c)(2+) response. Bafilomycin A and oligomycin had no effect on either the mitochondrial or cytoplasmic response. Inhibitors of both voltage regulated calcium channels and intracellular calcium release channels inhibited the Ca(2+)-dependent component of the Ca(mt)(2+) response to oxidative stress. We conclude that the more significant Ca(2+) response to oxidative stress occurs in the mitochondria and that both intracellular and extracellular calcium pools can contribute to the increases in [Ca(c)(2+)] and [Ca(mt)(2+)] induced by oxidative stress. PMID- 11882389 TI - Functional plasticity of cyclic AMP hydrolysis in rat adenohypophysial corticotroph cells. AB - Characterisation of cyclic nucleotide-hydrolysing phosphodiesterases (PDEs) in recombinant systems has highlighted regulatory properties indicative of distinct physiological roles for these enzymes. The present study investigated the role of PDEs in the adenosine 3'5'-monophosphate (cAMP) response to the hypothalamic neuropeptides corticotrophin-releasing factor (CRF) and arginine vasopressin (AVP) in acutely dispersed rat adenohypophysial cells. Ca(2+)-activated PDE (PDE1) and Ca(2+)-independent, rolipram-sensitive PDE (PDE4) accounted for close to 90% of cAMP-hydrolysing activity in the adenohypophysis. Messenger RNA transcripts of PDE1 (isotypes 1A and 1C) and PDE4 (isotypes B and D3) were detected by RT-PCR. The PDE blockers rolipram and IBMX enhanced cAMP accumulation induced by CRF or CRF and AVP. Vinpocetine, an inhibitor of low K(m) PDE1 isotypes, did not alter the response to CRF but enhanced the effect of the combined CRF/AVP stimulus. Thus, PDE4s terminate the cAMP response to moderate stimulation, while low-affinity PDE1 becomes important when the concentrations of CRF and AVP are characteristic of exposure to intensive stress. PMID- 11882390 TI - In addition to the SH3 binding region, multiple regions within the N-terminal noncatalytic portion of the cAMP-specific phosphodiesterase, PDE4A5, contribute to its intracellular targeting. AB - The long cyclic AMP (cAMP)-specific phosphodiesterase isoform, PDE4A5 (PDE4A subfamily isoform variant 5), when transiently expressed in COS-7 cells, was shown in subcellular fractionation studies to be associated with both membrane and cytosol fractions, with immunofluorescence analyses identifying PDE4A5 as associated both with ruffles at the cell margin and also at a distinct perinuclear localisation. Deletion of the first nine amino acids of PDE4A5 (1) ablated its ability to interact with the SH3 domain of the tyrosyl kinase, LYN; (2) reduced, but did not ablate, membrane association; and (3) disrupted the focus of PDE4A5 localisation within ruffles at the cell margin. This deleted region contained a Class I SH3 binding motif of similar sequence to those identified by screening a phage display library with the LYN-SH3 domain. Truncation to remove the PDE4A5 isoform-specific N-terminal region caused a further reduction in membrane association and ablated localisation at the cell margin. Progressive truncation to delete the PDE4A long isoform common region and then the long isoform-specific UCR1 did not cause any further change in membrane association or intracellular distribution. However, deletion up to the super short form splice junction generated an entirely soluble 'core' PDE4A species. We propose that multiple sites in the N-terminal noncatalytic portion of PDE4A5 have the potential to associate with intracellular structures and thus define its intracellular localisation. At least two such sites lie within the PDE4A5 isoform specific N-terminal region and these appear to be primarily responsible for targeting PDE4A5 to, and organising it within, the cell margin; one is an SH3 binding motif able to interact with LYN kinase and the other lies within the C terminal portion of the PDE4A5 unique region. A third membrane association region is located within the N-terminal portion of UCR2 and appears to be primarily responsible for targeting to the perinuclear region. Progressive N-terminal truncation, to delete defined regions of PDE4A5, identified activity changes occurring upon deletion of the SH3 binding site region and then upon deletion of the membrane association site region located within UCR2. This suggests that certain of these anchor sites may not only determine intracellular targeting but may also transduce regulatory effects on PDE4A5 activity. PMID- 11882391 TI - Identification of Drosophila Myt1 kinase and its role in Golgi during mitosis. AB - Entry into mitosis is regulated by inhibitory phosphorylation of cdc2/cyclin B, and these phosphorylations can be mediated by the Wee kinase family. Here, we present the identification of Drosophila Myt1 (dMyt1) kinase and examine the relationship of Myt1 and Wee1 activities in the context of cdc2 phosphorylation. dMyt1 kinase was found by BLAST-searching the complete Drosophila genome using the amino acid sequence of human Myt1 kinase. A single predicted polypeptide was identified that shared a 48% identity within the kinase domain with human and Xenopus Myt1. Consistent with its putative role as negative regulator of mitotic entry, overexpression of this protein in Drosophila S2 cells resulted in a reduced rate of cellular proliferation while the loss of expression via RNA interference (RNAi) resulted in an increased rate of proliferation. In addition, loss of dMyt1 alone or in combination with Drosophila Wee1 (dWee1) resulted in a reduction of cells in G2/M phase and an increase in G1 phase cells. Finally, loss of dMyt1 alone resulted in a significant reduction of phosphorylation of cdc2 on the threonine-14 (Thr-14) residue as expected. Surprisingly however, a reduction in the phosphorylation of cdc2 on the tyrosine-15 (Tyr-15) residue was only observed when both dMyt1 and dWee1 expression was reduced via RNAi and not by Wee1 alone. Most strikingly, in the absence of dMyt1, Golgi fragmentation during mitosis was incomplete. Our findings suggest that dMyt1 and dWee1 have distinct roles in the regulation of cdc2 phosphorylation and the regulation of mitotic events. PMID- 11882392 TI - Regulative potential of glutamine--relation to glutathione metabolism. AB - Glutamine (GLN) is the most abundant free amino acid (AA) in the human body. Under GLN-free conditions, which can be obtained when cells are cultivated in vitro, tissue cells cannot grow. Therefore, when classifying GLN as a "non essential" AA, one must consider that in the human body GLN is synthesized from essential AAs and is continuously delivered from skeletal muscle to other organs. It is fascinating that a relatively simple AA like GLN can stimulate a large variety of cellular reactions. GLN stimulates not only the growth of cells but also the expression of surface antigens, the formation of cytokines, and the synthesis of heat shock proteins. Further, a GLN deficiency leads to a cell cycle arrest in G(0) to G(1) and reduces apoptosis. Interestingly, many of these biological activities also are associated with the cellular reduced oxygen potential, which depends mainly on the ratio of reduced to oxidized glutathione. Experimental animal studies have shown that the administration of GLN increases tissue concentrations of reduced glutathione. This review describes the relation of GLN to reduced glutathione metabolism and discusses the alteration of reduced glutathione metabolism under a variety of clinical conditions such as reperfusion injury, myocardial infarction, respiratory insufficiency, cancer, diabetes, liver disease, and clinical protein catabolism. PMID- 11882393 TI - Glutamine: an anaplerotic precursor. AB - There is an up to four-fold increase in the concentration of the tricarboxylic acid (TCA) cycle intermediates at the start of exercise. The rate of TCA cycle flux and, hence, oxidative metabolism may be limited by the concentration of the intermediates in the cycle. The dramatic decline in intramuscular glutamate at the start of exercise, in tandem with increased intramuscular alanine, suggests that glutamate is an important anaplerotic precursor. We hypothesized that oral glutamine might enhance the exercise-induced TCA cycle intermediate pool expansion. Indeed, a greater increase in the sum of muscle citrate, malate, fumarate, and succinate concentrations (approximately 85% total TCA intermediate pool) occurred at the start of exercise after ingestion of glutamine rather than of placebo or ornithine alpha-ketoglutarate. However, neither endurance capacity nor the degree of phosphocreatine depletion or lactate accumulation was altered. This suggests that TCA cycle intermediates do not limit flux through the cycle or that more intense exercise is required to show the limitation. PMID- 11882394 TI - Glutamine and heat shock protein expression. AB - The most basic mechanism of cellular protection involves the expression of a highly conserved family of essential proteins, known as heat shock or stress proteins (HSPs). The expression of these proteins after a sublethal insult can induce "stress tolerance" and protect against a subsequent stress that otherwise would be lethal. Experimental data have shown that preinduction of the heat stress response can provide marked protection against many forms of cellular injury, including ischemia and reperfusion, lung injury, and shock. However, induction of HSPs to improve outcome in human disease has not been exploited because laboratory induction agents are themselves toxic and not clinically relevant. Many researchers have found that glutamine (GLN), a conditionally essential amino acid, can enhance stress-induced HSP expression in vitro and improve cell survival against a variety of stressful stimuli. Further, recent data from me and my colleagues indicate that a single dose of intravenous GLN can enhance HSP expression, decrease end-organ injury, and enhance survival from septic shock in the intact rat. Thus GLN, which is beneficial in many settings of critical illness and injury, may be a clinically applicable enhancer of HSP expression. These results indicate that GLN could be used to enhance HSP expression and attenuate end-organ injury in situations when a major clinical stress is anticipated, such as before major surgical procedures (e.g., cardiac, vascular, and transplantation) or in the critically ill. PMID- 11882395 TI - Prognostic value of energy metabolism in patients with viral liver cirrhosis. AB - The effect of energy malnutrition on survival in patients with non-alcoholic viral liver cirrhosis has not been well defined. We characterized energy metabolism at study entrance and prospectively analyzed its effect on subsequent survival in cirrhotics. One hundred nine consecutive patients with viral liver cirrhosis and 22 healthy control subjects participated in the study. By indirect calorimetry after overnight bedrest and fasting, resting energy expenditure (REE) was measured and non-protein respiratory quotient (npRQ) was calculated. Survival of cirrhotics were followed for up to 8 y. Survival rate was estimated with the Kaplan-Meier method. REE at entrance was significantly higher than the predicted basal metabolic rate (BMR) in cirrhotics (P < 0.001). NpRQ was significantly lower in cirrhotics than in controls (P < 0.001). Survival rate was significantly lower in patients with low npRQ ( < 0.85) than in patients with scores above 0.85 (P < 0.01) and was significantly higher in normal metabolic patients (0.9 < REE/BMR < 1.1) than in hypometabolic (REE/BMR < 0.9) or hypermetabolic (1.1 < REE/BMR) patients (P < 0.05). The proportional hazards model showed that npRQ (relative risk = 0.0003, 95% confidence interval = 0.0000-0.0970), REE/BMR (0.0199, 0.0007-0.5652), prothrombin time, and ammonia were independent significant factors determining survival. Thus evaluation of energy metabolism can be used to predict survival in patients with viral liver cirrhosis. PMID- 11882396 TI - Influences of soybean oil emulsion on stress response and cell-mediated immune function in moderately or severely stressed patients. AB - OBJECTIVES: We previously reported that omega-6 fat emulsion increases cytokine production in burned rats. Effects of soybean oil emulsion on surgical stress responses and lymphocyte function according to the surgical severity have not been studied in detail. We investigated the effects of soybean oil emulsion, which contains 50% omega-6 fatty acid, on postoperative stress responses and cell mediated immune function according to the severity of surgical stress. METHODS: Eight patients who underwent gastric or colorectal surgery and nine who underwent esophagectomy were fed fat-free total parenteral nutrition. Ten patients who underwent gastric or colorectal surgery and seven who underwent esophagectomy were fed total parenteral nutrition with soybean oil emulsion. Total parenteral nutrition provided 1.5 g of protein and 40 kcal per kilogram every day from 7 d before surgery to postoperative day 14. Soybean oil emulsion (Intralipid) accounted for 20% of the total calories. Serum interleukin-6, C-reactive protein, glucagon, and concanavalin A- or phytohemagglutinin-stimulated lymphocyte proliferation were determined. RESULTS: In the group of moderately stressed patients, soybean oil emulsion did not amplify the measured levels. In the group of severely stressed patients, soybean oil emulsion amplified the level of serum interleukin-6 and decreased concanavalin A- or phytohemagglutinin-stimulated lymphocyte proliferation. CONCLUSIONS: Soybean oil emulsion amplifies the stress responses and possibly suppresses cell-mediated immune function induced by surgical stress in severely stressed patients, but not in moderately stressed patients. PMID- 11882397 TI - Hypocaloric enteral tube feeding in critically ill obese patients. AB - OBJECTIVE: We respectively compared the nutritional and clinical efficacies of eucaloric and hypocaloric enteral feedings in 40 critically ill, obese patients admitted to the trauma or surgical intensive care unit. METHODS: Adult patients, 18 to 69 years old, with weights greater than 125% of ideal body weight, normal renal and hepatic functions, and who received at least 7 d of enteral tube feeding were studied. Patients were stratified according to feeding group: eucaloric feeding (>or=20 kcal/kg of adjusted weight per day; n = 12) or hypocaloric feeding (<20 kcal/kg of adjusted weight per day; n = 28). The goal protein intake for both groups was approximately 2 g/kg of ideal body weight per day. Clinical events and nutrition data were recorded for 4 wk. RESULTS: Patients were similar according to sex, age, weight, body mass index, Second Acute Physiology and Chronic Health Evaluation score, Trauma score, and Injury Severity Score. The hypocaloric feeding group received significantly fewer calories than the eucaloric group (P left atrial pressure). For further quantification mean gradients, time duration of pressure overlap, as well as products of mean gradients and overlap time were analysed. RESULTS: During the Valsalva manoeuvre a significant pressure gradient could be observed in 84% of the patients, followed by an expiration pressure of 60 mmHg (82%), inspiration (78%), expiration pressure of 40 mmHg (76%), coughing (75%) and an expiration pressure of 20 mmHg (62%). Comparing the mean gradients and the products of mean gradients and overlap time duration during the different manoeuvres, we could detect the significantly best results with the Valsalva manoeuvre. CONCLUSIONS: The Valsalva manoeuvre might be the most effective test to provoke a right-to left atrial shunt for the detection of a PFO during echocardiographic examinations. PMID- 11882434 TI - Tissue Doppler imaging in the evaluation of the regional diastolic function in chagas' disease. AB - AIMS: The main purpose of this study was to evaluate the regional diastolic function in Chagas' disease using tissue Doppler imaging (TDI). METHODS AND RESULTS: Seventy-seven patients were evaluated and divided into three groups: group 0, control; group 1, chagasic patients with normal ECG; and group 2, chagasic patients with abnormal ECG. The following parameters were evaluated: E and A waves, E/A ratio, deceleration time and the isovolumic relaxation time by pulsed wave Doppler and analysis of early (e') and late (a') filling waves by means of TDI, in basal segment of the septal, anterior, inferior, posterior and lateral walls of the left ventricle. The only index of the transmitral flow that presented a significant difference between the chagasic patients and the controls was the deceleration time. As regards the TDI, a significant difference occurred between the various groups in relation to the e' wave, especially in the septal, inferior and posterior walls of the left ventricle. CONCLUSION: TDI proved itself a more sensitive technique for the study of the diastolic function in Chagas' disease than conventional Doppler echocardiography, bringing to light regional abnormalities and allowing differentiation between normal and chagasic individuals with or without cardiomyopathy. TDI could be potentially useful in clinical work and in the stratification of risk of these patients. PMID- 11882435 TI - Images in echocardiography. A 70-year-old man presenting with heart failure. PMID- 11882436 TI - Full performance of modern echocardiography within the heart: in-vivo feasibility study with a new intracardiac, phased-array ultrasound-tipped catheter. AB - BACKGROUND: Intracardiac echocardiography with full performance of high resolution two-dimensional-, M-mode-, colour, pulsed and continuous wave Doppler and Doppler tissue imaging has not been previously demonstrated. AIMS: This first European in-vivo study was designed to determine the utility and feasibility of a new ultrasound-tipped catheter for intravascular and intracardiac echocardiography. METHODS: The miniaturized, multi-modal, multiple-frequency (5 10MHz) transducer tipped 10Fr (3.3mm) catheter was tested in five anaesthetized mongrel dogs linked to a standard echocardiographic platform. The catheter was introduced through an 11 Fr femoral venous sheath into the inferior vena cava and right heart chambers and the pulmonary artery under limited fluoroscopic and catheter ultrasound guidance. RESULTS: Abdominal and thoracic aorta as well as their branches, both ventricles and atrias with their appendices, all valves, pulmonary arteries and all veins could be visualized with excellent quality. All Doppler signals and the determined haemodynamics, global and regional wall motion and Doppler tissue imaging were of high diagnostic quality. Coronary flow reserve could also be determined. CONCLUSIONS: Intracardiac echocardiography is feasible and potentially useful for assessing functional and morphological disorders, and probably for the guidance of interventional procedures as well as monitoring of cardiac function. A new window to the heart has been opened. PMID- 11882437 TI - Doppler tissue echocardiography: myocardial wall motion velocities in essential hypertension. AB - AIMS: Doppler tissue echocardiography (DTE) was applied to extract the myocardial wall velocities along different planes and evaluate the left ventricular function in essential hypertension. METHODS AND RESULTS: Fifty-four hypertensives (HT) were compared to a control group of 31 normotensive (NT) subjects. The short-axis shortening and lengthening was assessed through the parasternal projections, sampling from interventricular septum and posterior wall. Through the apical projections the mitral annulus excursion was observed at four sites (anterior, posteroseptal, lateral, inferior walls) to assess the longitudinal dynamic of the heart. In each myocardial segment, peak velocity and time-velocity integral for systolic (S) and diastolic waves (E and A) were measured and their means for the long- and short-axis directions were calculated. Significant changes in hypertensives involved mainly the longitudinal motion. In diastole, the E-wave relaxation velocity was significantly decreased and the late A-wave velocity was unchanged. The E/A velocity ratio was significantly reduced. Relaxation velocity was negatively correlated to age, left ventricular mass and diastolic blood pressure. In systole, the peak S-wave shortening velocity was reduced and no association with age, left ventricular mass and blood pressure could be demonstrated. The range of segmental data produced by DTE proved useful to manufacture sensitive indices for recognition of hypertensive damage. Single DTE variables also proved slightly more sensitive than those extracted from the mitral flow pattern for the discrimination of HT patients. CONCLUSION: The presence of impaired relaxation was confirmed by DTE in a large portion of patients with hypertension and left ventricular hypertrophy. A peculiar systolic disturbance is evidenced by this technique. DTE-derived information can be used to detect early and quantify target-organ damage and its progression or regression during antihypertensive treatment. PMID- 11882438 TI - Are changes in left ventricular volume as measured with the biplane Simpson's method predominantly related to changes in its area or long axis in the prognostic evaluation of remodelling following a myocardial infarction? AB - AIMS: Two-dimensional (2D) echocardiography has been widely applied to measure left ventricular volumes with the biplane Simpson's method in the assessment of left ventricular remodelling following an acute myocardial infarction. This volume formula is based upon tracings of endocardium and measurement of long axis on left ventricular images. In the present follow-up study of post-myocardial infarction patients we evaluated the prognostic impact of changes in left ventricular areas and geometry versus long axis to determine if only long-axis measurements may be used for prognostic purposes. METHODS AND RESULTS: Two dimensional echocardiographic video recordings of the apical four-chamber and long-axis views were obtained in 756 patients 2--7 days and 3 months following an acute myocardial infarction. All videotapes were sent to a core laboratory and left ventricular volumes were measured with the biplane Simpson's method in end diastole and end-systole. During the first 3 months 44 patients had suffered one of the following end-points and were excluded: cardiac death, recurrent myocardial infarction, heart failure or chronic arrhythmia. Over a period of 3- 24 months 58 such end-points occurred. With the Cox proportional hazards model the increase in left ventricular systolic volume was the strongest predictor for such events (Chi-square 18.5, P<0.0001), followed by an increase in end-systolic area (Chi-square 17.0, P<0.0001) and end-systolic spherity index (Chi-square 8.74,P =0.003). The increase in end-systolic long axis had only a borderline predictive value (Chi-square 4.3, P=0.04). The change in long-axis shortening from end-diastole to end-systole had no significant predictive value at all. CONCLUSION: In the studied population changes in left ventricular area and geometry, but not in the long axis, were mainly related to cardiac morbidity. The proper assessment of changes in left ventricular dimensions should therefore be based upon tracings of the area and not on long axis measurements only. PMID- 11882439 TI - Left ventricular early diastolic inflow velocity and atrial ventricular plane downward velocity: useful parameters to test diastolic function in clinical practice? Diastolic parameters tested in a clinical setting. AB - AIMS: To study the clinical value of the colour-M-mode slope of the early diastolic left ventricular filling phase (Vp) and the early diastolic downward M mode slope of the left atrioventricular plane displacement (EDS), compared with diastolic function assessed by traditional Doppler evaluation. METHODS AND RESULTS: In 65 consecutive patients EDS and Vp were compared with a four-degree traditional diastolic function classification, based on pulsed Doppler assessment of the early to atrial transmitral flow ratio (E/A), the E-wave deceleration time (Edt), and the systolic to diastolic (S/D) pulmonary venous inflow ratio. Vp (P=0.006) and EDS (P=0.045) were related to traditional diastolic function (Kruskal--Wallis analysis). EDS showed a trend brake between the moderate and severe diastolic dysfunction groups by traditional Doppler evaluation. Vp and EDS correlated weakly in simple linear regression analysis (r=0.33). Vp and EDS discriminated poorly between normal and highly abnormal diastolic function. CONCLUSIONS: Vp and EDS were significantly related to diastolic function by traditional Doppler evaluation. They were, however, not useful as single parameters of left ventricular diastolic function due to a small difference between normal and highly abnormal values, allowing for little between measurement variability. Consequently, these methods for the evaluation of left ventricular diastolic function do not add significantly to traditional Doppler evaluation. PMID- 11882440 TI - Images in echocardiography. Exaggerated pectinate muscles mimicking multiple left atrial appendage thrombi. PMID- 11882441 TI - The stress echo prognostic gender gap. AB - AIMS: To investigate whether myocardial ischaemia elicitable during pharmacological stress echocardiography portends different prognosis in men and women. METHODS AND RESULTS: The study group was made by 1733 patients (941 men, 792 women) who underwent dipyridamole (n=1008) or dobutamine (n=725) stress echo for evaluation of known or suspected coronary artery disease. An ischaemic response was found in 460 patients (308 men, 152 women). Considering the whole ischaemic population, women were older (P<0.0001) and more likely to have hypertension (P=0.02) and hypercholesterolaemia (P=0.04) than men. No difference in age and risk factors was evidenced between the two sexes in the subset of 203 patients with ischaemia and suspected coronary artery disease. During follow-up (25 +/- 24 months for the ischaemic and 37 +/- 25 months for the non-ischaemic sample), there were 113 cardiac events (45 deaths and 68 infarctions) and 232 revascularizations. Revascularization rate in ischaemic population was similar in both sexes (P=0.36). Multivariate predictors of cardiac events in the whole ischaemic group were resting WMSI (HR=2.7, 95% CI 1.3--3.3;P=0.0050), female gender (HR=2.2, 95% CI 1.2--3.7;P=0.0062), age > or = l65 years (HR=1.9, 95% CI=1.0--3.6;P=0.0427), and Delta WMSI (HR=2.1, 95% CI=1.0--3.7;P=0.0447). Female gender (HR=2.7, 95% CI 1.1--6.3;P=0.0233) was the only independent prognostic predictor in patients with ischaemia and suspected coronary artery disease. Five year infarction-free survival was 82% in men and 71% in women in the whole ischaemic population (P=0.0041) as well as in the ischaemic group with suspected coronary artery disease (CAD) (P=0.0175). In the non-ischaemic sample resting WMSI (HR=4.8), history of myocardial infarction (HR=2.5), and hypercholesterolaemia (HR=1.8) were independent predictors of outcome at multivariate analysis, whilst the gender had no prognostic importance. CONCLUSIONS: Our results show that female gender is an independent predictor of cardiac events in patients with myocardial ischaemia induced by pharmacological stress echocardiography. PMID- 11882443 TI - Images in echocardiography. Isolated huge aneurysm of the non-coronary sinus of Valsalva. PMID- 11882442 TI - Thrombus in the main pulmonary artery of a patient with thromboangiitis obliterans: observation by transthoracic echocardiography. AB - We describe a 45-year-old man with thromboangiitis obliterans. He had a large immobile wall-adherent thrombus located in the main pulmonary artery, which was detected by transthoracic echocardiography. The pulmonary arterial involvement in this patient may suggest that thromboangiitis obliterans is a generalized vascular disease. We conclude that pulmonary artery should be thoroughly examined for thrombi in thromboangiitis obliterans patients who present with signs and symptoms of right heart failure. Transthoracic echocardiography should be the initial mode of examination in these patients. PMID- 11882444 TI - Echo contrast enhancement of the left ventricle--imaging technology and micro spheres, is the marriage critical? PMID- 11882445 TI - Diagnosis of major proximal coronary artery stenosis by transesophageal echo Doppler. PMID- 11882446 TI - Doppler myocardial imaging in the assessment of regional myocardial function in longitudinal direction pre- and post-PTCA. PMID- 11882447 TI - Pulmonary artery thrombi detected by echocardiography in patients with pulmonary hypertension secondary to atrial septal defect. AB - This report presents three patients with severe pulmonary hypertension secondary to atrial septal defect associated with thrombus and spontaneous echo contrast within the pulmonary artery diagnosed by transthoracic and transoesophageal echocardiography. Clinical and echocardiographic features seem to suggest local thrombus formation within the pulmonary arteries as a direct consequence of pulmonary hypertension rather than venous thromboembolism. PMID- 11882448 TI - For left ventricular opacification and endocardial border definition: is it really important which contrast agent we use, or is it the imaging modality we choose? AB - AIMS: The purpose of this study was to prospectively compare the effect of three imaging modalities (fundamental, harmonics, and power harmonics) on left ventricular opacification and endocardial border definition with two different echo agents, Optison and Albunex. METHODS: A total of 84 patients who had suboptimal transthoracic images were studied with echo contrast agents Albunex (n=41) and Optison (n=43). Each contrast agent was examined with three different imaging modalities, fundamental, harmonics and power harmonics, respectively. Left ventricular opacification was obtained by videodensitometric analysis. Percentage of endocardial border visualization was determined by indexing circumference of visualized endocardium to total circumference. Variables were compared with respect to three imaging modalities between two different echo agents. RESULTS: Higher videointensities and higher percentages of endocardial visualization were achieved with Optison compared to Albunex with fundamental and harmonics. However, there was no significant difference between Optison and Albunex with respect to LV opacification and border visualization by power harmonics. CONCLUSION: These results indicate that left ventricular opacification and endocardial border visualization is significantly improved by using power harmonic imaging as compared to harmonics or fundamental imaging following both echo contrast agents. Furthermore, although Optison is clearly superior to Albunex in opacifying left ventricle, power harmonic imaging compensates for the less robust agent. PMID- 11882449 TI - A semiquantitative method based on proximal convergence zone to estimate the severity of the mitral regurgitation: design and clinical application. AB - AIMS: The analysis of the mitral regurgitation using the proximal isovelocity surface area method has not been extended to clinical practice because of its complexity. Our objective was to design and validate a simplified semi quantitative method based on proximal isovelocity surface area to assess the severity of mitral regurgitation. METHODS AND RESULTS: We studied 58 patients with mitral regurgitation. We found a good correlation between the angiographic grade of mitral regurgitation and the parameters derived from proximal isovelocity surface area: maximal regurgitant flow, regurgitant orifice and volume (r: 0.90 to 0.92, P<0.05). The values of maximal regurgitant flow that best predicted the grades of angiographic severity (I--IV) were estimated by regurgitant orifice curves, with cut points of 16, 56 and 160ml.s( 1)kappa(p)=0.92). Considering that maximal regurgitant flow only depends on the radius of proximal isovelocity surface area and the velocity of aliasing used, we constructed a nomogram with the previously described limits. Twenty-four new patients were studied using this nomogram and angiography. An excellent degree of agreement was found (kappa(p)=0.93). The inter- and intraobserver variability showed a kappa(p)=0.89 and 0.91, respectively. CONCLUSION: This nomogram allows a fast semi-quantitative estimation of the grade of MR, feasible and highly correlative with the invasive methods. PMID- 11882450 TI - Diagnostics of main coronary artery stenoses and occlusions: multiplane transoesophageal Doppler echocardiographic assessment. AB - AIM AND METHODS: The possibility of using multiplane transoesophageal echocardiography (TEE) and quantitative coronary angiography (QCA) in the diagnostics of stenotic atherosclerosis of the main coronary arteries in a comparative aspect were studied in 94 patients with coronary artery disease (men, mean age 52 +/- 7 years). Coronary arteries stenoses were calculated with Doppler echocardiography using a modified continuity equation: stenosis (%)=100 x(1 prestenotic VTI(d)/stenotic VTI(d)) where prestenotic VTI(d), (cm)=diastolic velocity integral in the prestenotic zone, and stenotic VTI(d), (cm)=in the trans stenotic zone. RESULTS: High sensitivity and specificity of TEE in the diagnostics of stenotic and occlusive atherosclerosis of coronary arteries were revealed. They measured 88% and 98% for the left main coronary artery (LMCA), 97% and 67% for the left descending artery (LDA), 95% and 92% for the circumflex artery (CX), 83% and 97% for the right coronary artery (RCA), respectively. A high correlation was found between the results of TEE and QCA in the diagnostics of coronary stenoses which were made for the LMCA (r=0.82P <0.001), LDA (r=0.84, P<0.001), CX (r=0.85,P <0.001), and RCA (r=0.84, P<0.001). We developed Doppler echocardiography criteria for haemodynamically significant stenoses of coronary arteries (>50%) according to a peak diastolic velocity of the coronary blood flow, calculated as 1.4m.s(-1)for the LMCA, 0.9m.s(-1)for the LDA, and 1.1m.s( 1)for the CX. We determined Doppler echocardiography criteria of coronary arteries occlusions such as a 'break' of colour mapping, absence of Doppler spectrum and retrograde blood flow during late diastole. CONCLUSION: Transoesophageal Doppler evaluation of coronary blood flow with application of a modified continuity equation is an accurate, non-invasive method of coronary arteries stenoses diagnostics. PMID- 11882451 TI - Doppler myocardial imaging in the assessment of regional myocardial function in longitudinal direction pre- and post-PTCA. AB - AIMS: Doppler myocardial imaging is potentially a sensitive tool to assess regional myocardial velocities pre- and post-percutaneous transluminal coronary angioplasty (PTCA) as a marker of contractility to evaluate short- to medium-term information on functional myocardial recovery following the release of ischaemia. METHODS: Thirty patients with single vessel disease were studied to assess regional myocardial peak systolic velocity, systolic velocity time integral and mitral valve plane excursion in longitudinal direction one day pre-, one day post and 3 months post-PTCA. The patients were assigned to group A with coronary stenoses >70% and group B with stenoses < 70%. RESULTS: In group A pre-PTCA the ischaemic segments showed a significantly lower peak systolic velocity and velocity time integral compared with the values one day after PTCA (5.8 +/- 1.4 vs 7.7 +/- 1.4cm.s(-1); 1.06 +/- 0.22 vs 1.23 +/-0.28cm;P< 0.03). In contrast, mitral valve plane excursion in this group remained unchanged after PTCA for both the ischaemic and non-ischaemic left ventricular wall. In group B no changes of these parameters and no differences in mitral valve plane excursion of the ischaemic and the non-ischaemic left ventricular wall could be seen. CONCLUSION: With Doppler myocardial imaging it was possible to quantify a number of indices which changed due to the successful release of ischaemia. PMID- 11882452 TI - Myocardial performance after transmyocardial revascularization with CO(2)laser. A dobutamine stress echocardiographic study. AB - AIMS: Transmyocardial laser revascularization is a treatment for patients with severe angina pectoris not eligible for conventional revascularization. The effects on myocardial function and reversible ischaemia have not been clarified. METHODS AND RESULTS: One hundred patients with refractory angina not eligible for conventional revascularization were randomized 1:1 to receive continued optimal medical treatment or transmyocardial revascularization with CO(2)laser in addition to medical treatment. Dobutamine stress echocardiography examinations were performed at baseline and at 3 and 12 months after randomization. The effects of transmyocardial revascularization on myocardial function and reversible ischaemia were assessed by visual interpretation of cineloops at rest and during stress in a 16-segment model. After transmyocardial revascularization resting left ventricular wall motion abnormalities increased (P<0.01), whereas wall motion during dobutamine stimulation remained unchanged. The number of probably non-viable segments increased (P<0.01) with a corresponding decrease in the number of ischaemic segments. Fewer patients had the dobutamine infusion discontinued because of chest pain after transmyocardial revascularization with laser, but the chest pain threshold did not increase significantly. CONCLUSION: Following transmyocardial revascularization, resting wall motion abnormalities worsened, wall motion abnormalities during dobutamine stimulation remained unchanged and the number of probably non-viable segments increased. PMID- 11882453 TI - Tissue tracking allows rapid and accurate visual evaluation of left ventricular function. AB - AIMS: To evaluate the ability of tissue tracking for rapid assessment of left ventricular function by determination of the systolic mitral annular displacement. Tissue tracking is a new echocardiographic modality based on Doppler Tissue imaging allowing rapid visual assessment of the systolic baso apical displacement of each myocardial segment in apical views by a graded colour display. METHODS AND RESULTS: We studied 90 patients (69 male, age 60.4 +/- 10.1 years) with different left ventricular function (25 subjects with normal left ventricular function, 25 patients with homogeneous depression of left ventricular function and 40 patients with prior myocardial infarction). Systolic mitral annular displacement was determined by tissue tracking and M-mode echocardiography. Apical two-, three- and four-chamber views were used to determine the mitral annular displacement of six sites. Left ventricular ejection fraction was determined by two-dimensional echocardiography using Simpson's rule. Tissue tracking was possible in all patients. In the 50 patients with normal left ventricular function or homogeneous depression of left ventricular function, mean mitral annular displacement correlated closely with mitral annular displacement determined by M-mode (r=0.99,P <0.001) and with left ventricular ejection fraction (r=0.97, P<0.001). Left ventricular ejection fraction < or = 30% could be predicted with a sensitivity of 98% and a specificity of 78% using a cut-off value of 4.8mm for the mitral annular displacement determined by tissue tracking. In patients with prior myocardial infarction correlation between the mean mitral annular displacement and left ventricular ejection fraction was lower (r=0.87, P<0.001). CONCLUSION: Systolic mitral annular displacement determined by tissue tracking correlates closely with mitral annular displacement determined by M-mode and with left ventricular ejection fraction. Thus, tissue tracking allows rapid semiquantitative evaluation of global left ventricular function by assessment of systolic mitral annular displacement. PMID- 11882454 TI - Traumatic ventricular septal defect and mitral insufficiency after a Kebab's shish wound to the chest. AB - Transthoracic echocardiography has an important role in the assessment of patients with penetrating chest trauma. We report the case of 19-year-old boy who sustained a kebab's shish wound to the chest. Transthoracic echocardiography revealed a defect in the interventricular septum and a defect in the anterior mitral valve. Both of them were closed with direct sutures. PMID- 11882455 TI - Congenital diverticulum of the right ventricle associated with coarctation of aorta, atrial and ventricular septal defect and ductus. AB - Congenital right ventricular muscular diverticula are extremely rare and are usually associated with other congenital cardiac anomalies, (in half of the cases tetralogy of Fallot). They functionally behave like an accessory ventricular chamber which contracts synchronously with the normal ventricles. Less than 30 patients with a right ventricular diverticulum have been reported in literature. An apical right ventricular diverticulum occurs in patients with thoraco abdominal midline defects or abnormalities of the cardiac position([1]). However, an antero-superior diverticulum is usually associated with other congenital cardiac defects, such as a ventricular septal defect, tetralogy of Fallot, double outlet right ventricle and pulmonary stenosis([2--9]). We report an 11-year-old boy with an antero-superior diverticulum of the right ventricle associated with a coarctation of aorta, ductus arteriosus, and atrial and ventricular septum defects. To the best of our knowledge, such an association has not been reported before. PMID- 11882456 TI - Paradoxical cerebrovascular embolism associated with pulmonary arteriovenous fistula: contrast transoesophageal echocardiographic diagnosis. AB - We report two cases of paradoxical cerebrovascular embolism associated with intrapulmonary arteriovenous fistulas. In both cases the diagnosis was made by the use of contrast transoesophageal echocardiography, which not only detected the fistulas but also localized the arteriovenous fistula to specific pulmonary vascular beds. PMID- 11882457 TI - Mechanical strain, induced noninvasively in the high-frequency domain, is anabolic to cancellous bone, but not cortical bone. AB - Departing from the premise that it is the large-amplitude signals inherent to intense functional activity that define bone morphology, we propose that it is the far lower magnitude, high-frequency mechanical signals that continually barrage the skeleton during longer term activities such as standing, which regulate skeletal architecture. To examine this hypothesis, we proposed that brief exposure to slight elevations in these endogenous mechanical signals would suffice to increase bone mass in those bones subject to the stimulus. This was tested by exposing the hind limbs of adult female sheep (n = 9) to 20 min/day of low-level (0.3g), high-frequency (30 Hz) mechanical signals, sufficient to induce a peak of approximately 5 microstrain (micro epsilon) in the tibia. Following euthanasia, peripheral quantitative computed tomography (pQCT) was used to segregate the cortical shell from the trabecular envelope of the proximal femur, revealing a 34.2% increase in bone density in the experimental animals as compared with controls (p = 0.01). Histomorphometric examination of the femur supported these density measurements, with bone volume per total volume increasing by 32% (p = 0.04). This density increase was achieved by two separate strategies: trabecular spacing decreased by 36.1% (p = 0.02), whereas trabecular number increased by 45.6% (p = 0.01), indicating the formation of cancellous bone de novo. There were no significant differences in the radii of animals subject to the stimulus, indicating that the adaptive response was local rather than systemic. The anabolic potential of the signal was evident only in trabecular bone, and there were no differences, as measured by any assay, in the cortical bone. These data suggest that subtle mechanical signals generated during predominant activities such as posture may be potent determinants of skeletal morphology. Given that these strain levels are three orders of magnitude below strains that can damage bone tissue, we believe that a noninvasive stimulus based on this sensitivity has potential for treating skeletal complications such as osteoporosis. PMID- 11882458 TI - Cbfa1 does not regulate RANKL gene activity in stromal/osteoblastic cells. AB - The rates of osteoblast and osteoclast formation are tightly balanced, possibly due to the requirement of mesenchymal osteoblast progenitors for osteoclastogenesis. Osteoblast differentiation requires the transcription factor Cbfa1, whereas osteoclastogenesis results from the interaction between receptor activator of NF kappa B ligand (RANKL), expressed on stromal/osteoblastic cells, and RANK, a surface receptor on hematopoietic precursors. A striking decrease in the number of osteoclasts in Cbfa1-deficient mice suggested that Cbfa1 might be involved in RANKL expression. To investigate this possibility and to elucidate the mechanisms regulating RANKL expression, we isolated the 5'-flanking region of the murine RANKL gene and found that it contains two potential binding sites for Cbfa1 (OSE2-like sites). Cbfa1 bound to either of these sites in gel shift assays and stimulated the activity of a chimeric promoter consisting of multimerized RANKL OSE2-like sites inserted upstream from a minimal thymidine kinase (tk) promoter in transient transfections. However, Cbfa1 cotransfection did not stimulate murine RANKL promoter-luciferase constructs. Further analysis revealed that removal of these sites from the RANKL promoter by either site-directed mutagenesis or 5'-deletion did not alter the basal activity of promoter-reporter constructs. Conditional expression of Cbfa1 in a stromal/osteoblastic cell line stimulated osteocalcin mRNA by fivefold, but had no significant effect on RANKL mRNA levels. Conversely, conditional expression of a dominant-negative form of Cbfa1 in the same cell line inhibited osteocalcin mRNA by threefold, but had no effect on RANKL mRNA. Although these results cannot rule out a novel function for Cbfa1 in RANKL expression, they demonstrate that Cbfa1 does not regulate RANKL gene activity in the same manner as known targets of this transcription factor, such as osteocalcin. PMID- 11882459 TI - Discovery of osteoblast-associated genes using cDNA microarrays. AB - Osteoblast maturation is a complex process and involves distinct genotypic changes that are accompanied by specific phenotypic alterations. To identify new bone-related genes in osteoblasts we utilized the high-density mouse GEM1 microarray gene chip from IncyteGenomics, Inc. (St. Louis, Mo). We examined the expression profiles of over 8700 genes during the proliferation (day 3) and the mineralization (day 34) phases of MC3T3-E1 development. More than 8600 genes provided measurable signals. Of these genes, 252 were found to be differentially expressed on days 3 and 34. A large number of these genes have never been previously recognized in the context of osteoblast development. Approximately, 60% of the genes with expressions that were dominant in proliferating osteoblasts consisted of growth-related genes such as TACC3 and Pr22. The expressions of TIS21/BTG2, and a novel gene EST350, were found to peak during the differentiation phase (day 12), suggesting that they may play important roles in osteoblast differentiation. The majority of the genes with expressions that were dominant during the mineralization phase consisted of signal transduction genes and extracellular matrix (ECM) proteins such as lumican and cystatin-C. It is significant that lumican expression could not be detected on day 3, which indicates that this gene may serve as an important marker of postmitotic osteoblasts. The establishment of the expression profiles of these and other genes with various phases of MC3T3-E1 osteoblast development will allow us to distinguish the molecular events at different phases of osteoblast biology. PMID- 11882461 TI - Expression of estrogen receptor alpha and beta in the epiphyseal plate of the rat. AB - In this study we examine the spatial and temporal expression of estrogen receptor (ER) alpha and beta mRNA and protein in the tibial growth plate of the rat after birth, as well as the hormonal regulation of their expression. Using in situ hybridization and immunohistochemistry, we demonstrated ER alpha and ER beta mRNA and protein in tibial growth plates from 1 to 40 weeks after birth. ER alpha and beta mRNA and protein were localized in late proliferating and early hypertrophic chondrocytes during early life (1 and 4 weeks of age), whereas the immunohistochemistry also showed staining for ER alpha and beta in the resting cells. A similar expression pattern was observed during sexual maturation (7 weeks of age) except that ER beta mRNA was also detected in early proliferating chondrocytes. After sexual maturation (from 12 up to 40 weeks of age) ER alpha and beta mRNA and protein expression was confined to late proliferating and early hypertrophic chondrocytes. Apart from a relatively higher ER alpha mRNA expression in males after sexual maturation, we did not detect differences in expression of ERs between genders. Expression of ER beta mRNA in epiphyseal plates was increased in growth-retarded hypophysectomized rats compared with controls. Administration of growth hormone (GH) did not reverse the increased ER expression to normal. These data suggest that ER alpha and beta are coexpressed in growth plates of the rat after birth and that the level of expression of ERs in these tissues is hormonally regulated. Furthermore, our data indicate that the absence of growth-plate closure in the rat cannot be explained by disappearance of ER alpha expression during sexual maturation per se. PMID- 11882460 TI - Vascular endothelial growth factor stimulates chemotactic migration of primary human osteoblasts. AB - Recent studies have indicated a critical role for vascular endothelial growth factor (VEGF) during the process of endochondral ossification, in particular in coupling cartilage resorption with bone formation. Therefore, we studied the chemoattractive and proliferative properties of human VEGF-A on primary human osteoblasts (PHO) and compared these data with the effects of human basic fibroblast growth factor (bFGF) and human bone morphogenetic protein-2 (BMP-2). Furthermore, initial experiments were carried out to characterize VEGF-binding proteins on osteoblastic cells possibly involved in the response. For the first time, to our knowledge, we could demonstrate a chemoattractive effect of VEGF-A, but not VEGF-E, on primary human osteoblasts. The effect of VEGF-A was dose dependent and did not reach a maximum within the concentration range tested (up to 10 ng/mL). The maximal effect observed was a chemotactic index (CI) of 2 at a concentration of 10 ng/mL. bFGF and BMP-2 exhibited maxima at 1.0 ng/mL with CI values of 2.5 and 2, respectively. In addition to its effect on cell migration, VEGF-A stimulated cell proliferation by up to 70%. Reverse transcription polymerase chain reaction (RT-PCR) analysis revealed the expression of VEGF receptors VEGFR-1 (Flt-1), VEGFR-2 (Kdr), and VEGFR-3 (Flt-4), as well as neuropilin-1 and -2. An in vitro kinase assay failed to demonstrate activation of VEGFR-2 upon stimulation with either VEGF-E or VEGF-A, consistent with the idea that the effect of VEGF-A on primary human osteoblasts is mediated via VEGFR-1. Taken together, our data establish that human osteoblasts respond to VEGF-A, suggesting a functional role for this growth factor in bone formation and remodeling. PMID- 11882462 TI - Regulation of bone volume is different in the metaphyses of the femur and vertebra of C3H/HeJ and C57BL/6J mice. AB - The C3H/HeJ (C3H) mice exhibited a greater bone formation rate (BFR) and a greater mineral apposition rate (MAR) in the cortical bone of the midshafts of the femur and tibia than did C57BL/6J (B6) mice. This study sought to determine if these strain-related differences would also be observed in cancellous bone. Metaphyses of the femur and lumbar vertebra (L5-6) from C3H and B6 mice, 6 and 12 weeks of age, were analyzed by histomorphometry. Similar to cortical bone, the bone volume in the femoral metaphysis of C3H mice was greater (by 54% and 65%, respectively) than that of B6 mice at both 6 and 12 weeks of age. Higher BFR and mineral apposition rate (MAR) contributed to the higher bone volume in the C3H mice compared with the B6 mice. In contrast, bone volume (by 59% and 13%, respectively, p < 0.001) and trabecular number (by 55% and 35%, respectively, p < 0.001) in the vertebrae were lower in the C3H mice than in B6 mice at 6 and 12 weeks of age. At 6 weeks of age, MAR was higher (by 43%, p = 0.004) in C3H mice, but because of a low trabecular number, the BFR (by 37%, p = 0.026) and tetracycline-labeled bone surface (by 52%, p < 0.001) per tissue were lower in the vertebrae of C3H mice than B6 mice. The low bone volume in vertebrae of C3H mice was probably not due to a higher bone resorption, because the osteoclast number (by 55%, p < 0.001) and eroded surface (by 61%, p <0.001) per tissue area in the C3H mice were also lower in B6 mice. At 12 weeks, the trabecular thickness had increased (by 36%, p < 0.001) in the C3H mice and the difference in bone volume between strains was less than that at 6 weeks. These contrasting and apparently opposing strain-related differences in trabecular bone parameters between femur and vertebra in these two mouse strains suggest that the genetic regulation of bone volume in the metaphyses of different skeletal sites is different between C3H and B6 mice. PMID- 11882463 TI - Characterization of bone mineral composition in the proximal tibia of cynomolgus monkeys: effect of ovariectomy and nandrolone decanoate treatment. AB - Life postmenopausal women, ovariectomized cynomolgus monkeys (Macaca fascicularis) experience accelerated loss of bone mass. Treatment of ovariectomized monkeys with nandrolone decanoate results in an increase in bone mass to levels comparable to those of intact animals. The changes in bone composition that occur with these treatments, however, are less well characterized. In the present study, we used synchrotron Fourier-transform infrared microspectroscopy (FT-IRM) and curve-fitting methods to monitor specific changes at cortical, subchondral, and trabecular bone regions in the proximal tibia. Four groups were studied: (1) sham-operated (sham); (2) ovariectomized and treated with placebo for 2 years (ovx); (3) ovx + nandrolone decanoate for 2 years (NAN); and (4) ovx + nandrolone decanoate beginning 1 year after ovx (dNAN). The results demonstrate that ovariectomy and nandrolone treatment did not affect the degree of mineralization as defined by the phosphate/protein ratio, but acid phosphate content (HPO(4)(2-)) in cortical and subchondral bone was increased by ovariectomy, suggesting this bone to be less mature due to increased remodeling that occurs after ovariectomy. In the subchondral and cortical bone regions, ovariectomized monkeys showed a lower total carbonate content (CO(3)(2 )/matrix ratio) than sham controls, specifically due to the decrease in labile carbonate content. In the trabecular region, no change of carbonate content was observed. Treatment with nandrolone decanoate was found to restore the loss in carbonate, where the resulting mineral had a larger quantity of type B carbonate. Finally, we correlated carbonate content with dual-energy X-ray absorptiometry measurements, and found a positive correlation between bone mineral density and type A carbonate in bone, which is stoichiometrically related to the amount of calcium in bone. Therefore, the results presented herein identify significant differences in bone chemistry after ovariectomy and nandrolone treatment, which may help explain previous findings that, although nandrolone decanoate treatment increased bone mass, it could not reverse the decrease in bone strength due to ovariectomy. PMID- 11882464 TI - Syndrome of myelofibrosis and osteosclerosis: a series of case reports and review of the literature. AB - Myelofibrosis (MF) is a chronic disorder characterized by bone marrow evidence of myeloid metaplasia associated with reactive fibrosis, angiogenesis, and osteosclerosis. We report serum biochemistry, noninvasive markers of bone turnover, tetracycline-labeled bone histomorphometry, and bone densitometry (DXA) data of four men presenting with newly diagnosed biopsy-proven MF and osteosclerosis. The mechanisms and putative growth factors responsible for this syndrome are examined. PMID- 11882465 TI - Predicting human vertebral bone strength by vertebral static histomorphometry. AB - The study investigates the relationship between static histomorphometry and bone strength of human lumbar vertebral bone. The ability of vertebral histomorphometry to predict vertebral bone strength was compared with that of vertebral densitometry, and also with histomorphometry and bone strength of iliac crest bone biopsies. The material comprised matched sets of second lumbar vertebrae, third lumbar vertebrae, and two iliac crest bone biopsies from each of 21 women (19--96 years) and 24 men (23--95 years). One of the iliac crest biopsies and 9-mm-thick mediolateral slices of half of each of the entire vertebral bodies (L-2) were used for histomorphometry. The other iliac crest biopsies and the L-3 were destructively tested by compression. High correlation was found between BV/TV or Tb.Sp and vertebral bone strength (absolute value of r = 0.86 in both cases). Addition of Tb.Th significantly improved the correlation between BV/TV and bone strength, and the addition of bone space star volume significantly improved the correlation between Tb.Sp and bone strength (from absolute value of r = 0.86 to absolute value of r = 0.89 in both cases). Bone structure (connectivity density) was not capable of improving the prediction of bone strength of the vertebral body. The correlations between BV/TV of L-2 and bone strength of L-3 were comparable with the correlation obtained by quantitative computed tomography (QCT), peripheral QCT (pQCT), and dual-energy X ray absorptrometry (DEXA) of L-3 and bone strength of L-3. The iliac crest was found to have low predictive power of vertebral bone strength (iliac BV/TV: r = 0.62; iliac bone strength: r = 0.67). No gender-related differences were found in any of the relationships. It was shown that trabecular bone volume BV/TV and mean trabecular plate separation Tb.Sp are good predictors of vertebral bone strength. The ability of histomorphometry to predict vertebral bone strength was comparable to that of densitometry. Bone structure assessed by connectivity density did not improve the correlation between static histomorphometric measures and vertebral bone strength. No gender-related differences were found in any of the relationships. Neither static histomorphometry nor biomechanical testing of iliac crest bone biopsies is a good predictor of vertebral bone strength. PMID- 11882466 TI - Fatigue damage-fracture mechanics interaction in cortical bone. AB - Fatigue loading causes accumulation of damage that may lead to the initiation of a macrocrack and result in a catastrophic failure of bone. The objective of this study was to examine the influence of fatigue damage on crack growth parameters in bovine cortical bone. Nineteen rectangular beam specimens (4 x 4 x 48 mm) were machined from bovine tibiae. The long axis of the beams was aligned with the long axis of bones. Using a four-point bending fatigue setup, ten specimens were fatigue-damaged to different levels as indicated by stiffness loss. A through thickness notch was machined at the center of each damaged and undamaged beam. The notched specimens were then monotonically loaded beyond failure using a three point bending protocol. Critical stress intensity factor, K(I), and work to critical load, W(Q), were significantly lower in the damaged group than in the undamaged group (p < 0.03). When the undamaged specimens were assigned a percent stiffness loss of zero and pooled with the damaged group, significant negative correlations of percent stiffness loss with K(I) (R = 0.58, p < 0.01), W(Q) (R = 0.54, p < 0.02), maximum load, P(max) (R = 0.59, p < 0.008), deflection at maximum load, Delta(max) (R = 0.48, p < 0.04), structural stiffness, S(max) (R = 0.53, p < 0.02), W(max) (R = 0.55, p < 0.02), and load at 1.4 mm deflection (a value beyond failure but without complete fracture), P(1.4) (R = 0.47, p < 0.05), were found. Post hoc analysis revealed that the average load-deflection curve from the damaged group was transformable into that from the undamaged group through a special shift on the load-deflection plane. Fatigue damage reduces bone stiffness and resistance to crack initiation, maximum load-carrying capacity, and deflection before and after failure in cortical bone. The data suggest there is a single rule that governs the overall effect of fatigue damage on the fracture behavior of cortical bone. PMID- 11882467 TI - Lower extremity muscle strength does not independently predict proximal femur bone mineral density in healthy older adults. AB - The relationship described in the published literature between muscle strength and bone mineral density of older adults is not entirely certain. It is possible that the direct relationship reported in some studies is biased by failing to mathematically account for the biological influence of body weight and body height on both bone mineral density and muscle strength. This study sought to determine if the relationships between measures of lower extremity muscle strength and bone mineral density of the proximal femur are independent of body size (i.e., body height and body weight) in healthy older adults. We recruited 50 older women and 29 older men, all of whom were healthy community dwellers and not involved in resistance training. Quantitative analysis of the isometric strength of the bilateral ankle, knee, and hip joints and assessment of bone mineral density of the proximal femur were conducted. Muscle strength values were adjusted for the influence of body height and body weight using an allometric scaling procedure. The correlations between proximal femur bone mineral density and the unadjusted strength values were weak but statistically significant. After adjusting muscle strength to account for the influence of body height and body weight, the magnitudes of the correlations between bone mineral density and muscle strength diminished substantially and were not significantly different from zero. The results reveal that, for a typical sample of healthy older adults not involved in resistance training, the relationship between maximal isometric muscle strength of lower extremity joints and proximal femur bone mineral density is reliant on body size. PMID- 11882468 TI - Milk consumption and bone mineral content in Chinese adolescent girls. AB - A cross-sectional study of a random sample of 649 girls, aged 12-14 years (mean +/- SD: 12.9 +/- 0.6 years), in the Beijing area examined the relationship between diet and bone mineral status. Food and nutrient intakes over the past year were estimated by means of a semiquantitative food frequency questionnaire. Bone mineral content (BMC) and bone width (BW) at the distal one-third and one tenth radius and ulna were measured by single-photon absorptiometry. Results showed Beijing pubertal girls had a low mean milk consumption (fresh and powdered milk, vitamin D-fortified milk, and yogurt) at 50 g/day (95% confidence interval [CI] 44-55 g/day whereas one-third consumed no milk at all. Mean calcium intake was 356 +/- 97 mg/day of which only 21% was provided by milk and milk products. Milk intake varied by region (rural, suburban, and urban: 9, 36, and 83 g/day, respectively, p < 0.0005) as did the proportion of milk consumers in the three areas (30%, 64%, and 91%, p < 0.0005). Bone mineral density (BMD) at the distal one-third and one-tenth radius and one-tenth ulna was positively associated with milk consumption (p < 0.05). Multiple regression analysis of BMC on foods and nutrients as well as confounding factors, including weight, bone age, Tanner stage, and School Physical Activity Score (SPAS), showed that milk intake was the only dietary factor included in the models for BMC at the four bone sites measured. The model explained 54%-65% of the variation in BMC, and milk alone accounted for up to 3.2% of the variation. Milk was the only food group with significant partial correlation with BMC. SPAS, weight, bone age, and Tanner stage each accounted for a smaller variation in BMC (<1.8%). The results indicate that milk (presumably as an integrated source of nutrients) had a beneficial effect on bone mass of Beijing pubertal girls and was a better nutritional determinant of BMC than intake of any milk nutrient alone. Promotion of milk consumption should be considered for achieving optimal bone mass in this population group. PMID- 11882469 TI - Appraisal of knowledge and attitude of Akwa Ibomites toward a sustainable environment in Nigeria. AB - Developing countries are constantly and continuously confronted with a myriad of health and environmental problems that threaten the efforts to establish and maintain safe air and water supplies. Nigeria, in general, and Akwa Ibom State, in particular, are not isolated from increasing health and sanitation problems. My goal in this study was to determine the knowledge base of men and women related to sanitation and environmental health in Akwa Ibom State. I found no differences in the attitudes of men and women toward the sustainability of health or a healthy environment in the state. However, more sanitation and environmental education is needed as part of long-term strategy for abating sanitation and environmental health problems in this state. PMID- 11882470 TI - Impact of current good manufacturing practices and emission regulations and guidances on the discharge of pharmaceutical chemicals into the environment from manufacturing, use, and disposal. AB - The current Good Manufacturing Practice (cGMP) and effluent emission (use and disposal) regulations of the U.S. Food and Drug Administration (FDA) and manufacturing effluent discharge and emission regulations of the U.S. Environmental Protection Agency (U.S. EPA) require contained manufacture, use, and disposal of pharmaceuticals with the goal of minimizing the release of pharmaceutical chemicals into the environment. However, debate has recently arisen in several scientific forums over whether these regulations adequately protect human and environmental health from the new pharmaceutical drugs introduced each year into the marketplace and the multitude of existing products, each with many distinct biochemical modes of actions. To address this issue, it is important to understand the relevance of current cGMP regulations and emission regulations that have a direct bearing on the releases of pharmaceutical chemicals into the environment during the manufacture, use, and disposal of active pharmaceutical ingredients (drug substances) and drug products. This knowledge may help us assess the quantity of residues that may be released into the environment. Additionally, the information on physical, chemical, and degradation and sorption properties of the pharmaceutical chemicals may help determine the net residue levels that could persist in the environment to evaluate if such residues have any bearing on human and environmental health. The scientific and regulatory aspects of issues related to the manufacture, use, and disposal of pharmaceutical chemicals are discussed in this article, with special emphasis on potential environmental exposure pathways during the life cycle of an active pharmaceutical ingredient or drug product. The mechanisms of degradation (transformation or depletion) and dilution of pharmaceutical residues that may be released into aquatic or terrestrial environmental compartments are described. Such degradation and dilution of pharmaceutical chemicals in the environment may significantly reduce the residues. It is important to evaluate whether such residue levels have any measurable impact on human and/or environmental health. PMID- 11882471 TI - A combined analysis of the short-term effects of photochemical air pollutants on mortality within the EMECAM project. AB - In recent years, some epidemiologic studies have attributed adverse effects of air pollutants on health not only to particles and sulfur dioxide but also to photochemical air pollutants (nitrogen dioxide and ozone). The effects are usually small, leading to some inconsistencies in the results of the studies. Furthermore, the different methodologic approaches of the studies used has made it difficult to derive generic conclusions. We provide here a quantitative summary of the short-term effects of photochemical air pollutants on mortality in seven Spanish cities involved in the EMECAM project, using generalized additive models from analyses of single and multiple pollutants. Nitrogen dioxide and ozone data were provided by seven EMECAM cities (Barcelona, Gijon, Huelva, Madrid, Oviedo, Seville, and Valencia). Mortality indicators included daily total mortality from all causes excluding external causes, daily cardiovascular mortality, and daily respiratory mortality. Individual estimates, obtained from city-specific generalized additive Poisson autoregressive models, were combined by means of fixed effects models and, if significant heterogeneity among local estimates was found, also by random effects models. Significant positive associations were found between daily mortality (all causes and cardiovascular) and NO(2), once the rest of air pollutants were taken into account. A 10 microg/m(3) increase in the 24-hr average 1-day NO(2)level was associated with an increase in the daily number of deaths of 0.43% [95% confidence interval (CI), 0.003-0.86%] for all causes excluding external. In the case of significant relationships, relative risks for cause-specific mortality were nearly twice as much as that for total mortality for all the photochemical pollutants. Ozone was independently related only to cardiovascular daily mortality. No independent statistically significant relationship between photochemical air pollutants and respiratory mortality was found. The results in this study suggest that, given the present levels of photochemical pollutants, people living in Spanish cities are exposed to health risks derived from air pollution. PMID- 11882472 TI - Environmental organochlorines and semen quality: results of a pilot study. AB - There have been numerous studies that suggest that sperm concentrations (sperm counts) are declining in men. However, other studies suggest that sperm counts are not declining or may be increasing in some areas. Although there is disagreement on whether there is a downward temporal trend in sperm counts, the studies provide evidence that sperm counts vary by geographic location. It has been hypothesized that the geographic variation in sperm concentrations may be due to environmental exposures, lifestyle factors, or some unknown causes. To determine whether contemporary ambient levels of polychlorinated biphenyls (PCBs) and p,p-DDE are associated with altered semen quantity and quality, we selected a study population without specific exposure to PCBs or p,p-DDE. The present study presents the results from a pilot study on the relationship between serum PCBs and p,p-DDE and semen quality in 29 subjects recruited from the Massachusetts General Hospital Andrology Laboratory. Of the 29 subjects, 3 had sperm concentrations < 20 million/mL, 7 had < 50% motile sperm, 9 had < 4% normal morphology, and 6 were below normal in more than one semen parameter. The 18 subjects with normal spermatozoa concentration, motility, and morphology were used as comparison subjects. The mean (SE) concentration of the sum of PCBs and p,p-DDE was 242 ng/g lipids (34.0) and 354 ng/g lipids (120), respectively, for men with below normal motility as compared to 202 ng/g lipids (16.6) and 240 ng/g lipids (31.1), respectively, for the comparison subjects. The data showed general trends that were suggestive of an association between PCBs and p,p-DDE and abnormal motility, as well as with sperm concentration and morphology. A full scale study is currently in progress. PMID- 11882473 TI - Analysis of aggregate exposure to chlorpyrifos in the NHEXAS-Maryland investigation. AB - As part of the National Human Exposure Assessment Survey (NHEXAS) in Maryland, we collected indoor air, carpet dust, exterior soil, and duplicate diet samples from a stratified random sample of 80 individuals to evaluate aggregate daily exposure, contributions of specific pathways of exposure, and temporal variation in exposure to chlorpyrifos. We collected samples from each participant in up to six equally spaced sampling cycles over a year and analyzed them for chlorpyrifos. We used chlorpyrifos concentrations in each medium and self reported rates of time spent inside at home, time and frequency of contact with carpet, frequency of contact with soil, and weights of the duplicate diet samples to derive exposure to chlorpyrifos from each medium as well as average daily aggregate exposure (nanograms per day). The mean aggregate daily exposure to chlorpyrifos of 36 measurements obtained from 31 people was 1,390 ng/day (SD, 2,770 ng/day). Exposure from inhalation of indoor air accounted for 84.7% of aggregate daily exposure to chlorpyrifos on average. Chlorpyrifos concentrations in indoor air and carpet dust and the corresponding exposure rates were significantly correlated. Repeated short-term measurements of chlorpyrifos in carpet dust from individual residences were strongly correlated over time (reliability coefficient, R = 0.90), whereas the short-term measurements of chlorpyrifos in indoor air (R = 0.55) and solid food (R = 0.03) had moderate to weak reliability. Exposure to chlorpyrifos through those media and in aggregate based on direct measurements reported in this study can be used to understand better the accuracy of pesticide safety assessments. PMID- 11882474 TI - Circulating testosterone and prostate-specific antigen in nipple aspirate fluid and tissue are associated with breast cancer. AB - Preliminary evidence has associated testosterone and prostate-specific antigen (PSA) with breast cancer. Our objective was to determine whether a) testosterone levels in nipple aspirate fluid (NAF), serum, or breast tissue are associated with breast cancer; b) testosterone levels in serum are associated with levels in NAF; c) PSA in NAF, serum, or breast tissue is associated with breast cancer; and d) serum PSA is associated with NAF PSA levels. We obtained 342 NAF specimens from 171 women by means of a modified breast pump. Additionally, we collected 201 blood samples from 99 women and 51 tissue samples from 41 subjects who underwent surgical resection for suspected disease. Women currently using birth control pills or hormone replacement therapy were excluded from the study. Controlling for age and menopausal status, serum testosterone was significantly increased in women with breast cancer (p = 0.002). NAF and serum testosterone levels were not associated. Neither NAF nor tissue testosterone was associated with breast cancer. Controlling for menopausal status and age, NAF PSA was significantly decreased in women with breast cancer (p < 0.001). We did not find serum PSA to be associated with breast cancer, although we found an indication that, in postmenopausal women, its levels were lower in women with cancer. Serum PSA was associated with NAF PSA in postmenopausal women (p < 0.001). PSA levels in cancerous tissue were significantly lower than in benign breast specimens from subjects without cancer (p = 0.011), whereas levels of PSA in histologically benign specimens from subjects with cancer were intermediate. Our results suggest that serum testosterone is increased and NAF PSA is decreased in women with breast cancer, with PSA expression being higher in normal than in cancerous breast tissues. NAF and serum PSA levels in postmenopausal women are correlated, suggesting that as laboratory assessment of PSA becomes more sensitive, serum PSA may become useful in identifying women with breast cancer. PMID- 11882475 TI - Sexually dimorphic behavioral responses to prenatal dioxin exposure. AB - Pregnant Sprague-Dawley rats received a single oral dose of 0, 20, 60, or 180 ng/kg 2,3,7,8-tetrachlorodibenzo-p-dioxin on day 8 of gestation. Each litter contributed a single male-female pair trained to press a lever to obtain food pellets under two operant behavior procedures. Initially, each lever press was reinforced. The fixed-ratio (FR) requirement was then increased every four sessions from the initial setting of 1 to values between 6 and 71. We then studied responses for 30 days under a multiple schedule combining FR 11 and another schedule requiring a pause of at least 10 sec between responses (DRL 10 sec). TCDD evoked a sexually dimorphic response pattern. Generally, TCDD-exposed males responded at lower rates than control males. In contrast, exposed females responded at higher rates than controls. Each response measure from the mult-FR DRL schedule yielded a male-female difference score. We used the differences in response rate to calculate benchmark doses based on the relative displacement from modeled zero-dose performance of the effective dose at 1% (ED(01)) and 10% (ED(10)), as determined by a second-order polynomial fit to the dose-effect function. For the male-female difference in FR rate of responding, the mean ED(10) was 2.77 ng/kg with a 95% lower bound of 1.81 ng/kg. The corresponding ED(01) was 0.27 ng/kg with a 95% lower bound of 0.18 ng/kg. For the male-female difference in DRL rate, the mean ED(10) was 2.97 ng/kg with a 95% lower bound of 2.02 ng/kg. The corresponding ED(01) was 0.30 ng/kg with a 95% lower bound of 0.20 ng/kg. These values fall close to, but below, current estimates of human body burdens of 13 ng/kg, based on TCDD toxic equivalents. PMID- 11882476 TI - Partitioning of benzene in blood: influence of hemoglobin type in humans and animals. AB - Earlier studies have shown that air/blood partition coefficients (PCs) for many volatile organic chemicals (VOCs) are much higher in rat blood than in human blood. It has been suggested that the discrepancy could be attributed to the fact that hemoglobin (Hb) in rat blood exists in a quasi-crystalline form of hydrophobicity greater than that of normal human Hb (HbA) and thus has a higher carrying capacity for VOCs. In the present study, we used benzene as a prototypic VOC to examine its relative partitioning into human and animal blood. Additionally, we sought to ascertain whether the water-insoluble form of hemoglobin (HbS) found in subjects with homozygous sickle cell (SC) disease has a greater VOC-carrying capacity than does HbA blood. At a low-O(2) tension, HbS switches to water-insoluble polymers, which physically deforms the red blood cells (RBCs) to the sickle shape. We equilibrated HbA, HbS, Hartley guinea pig, CD1 mouse, and rat (F-344, Wistar, and Sprague-Dawley) blood and their respective fractions with benzene vapor (80 or 400 ppm) for 3 hr at 37 degrees C in air tight vials. We introduced benzene vapor into the vial head space that contained air or respiratory mixtures of venous-type (low-O(2)) or arterial-type (high O(2)) gases. The blood measurements included the PC, Hb, partial pressures of O(2)(pO(2)) and CO(2)(pCO(2), pH, and percentage of SCs. The benzene concentration had no effect on these parameters, and the high- and low-O(2) gas mixtures produced the expected changes in pO(2), pCO(2), and pH. At equilibrium, the low-O(2) HbS blood had approximately 85% SCs compared with roughly 15% with air or high-O(2) gas. PCs for rat and mouse blood were about 100% higher than those for human and guinea pig blood, but the PC for deoxygenated HbS blood was only slightly higher than that for HbA or oxygenated HbS blood. Benzene showed higher affinities for RBCs in the deoxygenated HbS, rat, and mouse blood and higher affinity for plasma in the guinea pig blood. There was no evidence of disproportionate partitioning of benzene into oxygenated HbS or into HbA blood forms. These data suggest that the water solubility of Hb alone appears to have little effect on the VOC-carrying capacity of blood and that the influence of species is large in comparison. These latter differences in partitioning may depend on the number of hydrophobic sites on the surface of the plasma/heme proteins and thus be unique to the species. PMID- 11882477 TI - The relationship between lead in plasma and whole blood in women. AB - Studies have suggested that plasma lead levels may better reflect the toxicologically labile fraction of circulatory Pb that is more freely available for exchange with target tissues than do Pb levels in whole blood. Studies have also reported an apparent severalfold variation in the relative partitioning of Pb between whole blood and plasma (or serum) for a given whole-blood Pb level. This may reflect inherent differences in the plasma Pb/whole blood Pb partitioning among individuals and/or methodologic challenges associated with the collection and analyses of samples that generally contain < 1-2 ng total Pb. Here, we conducted a longitudinal assessment of the relationship between Pb in whole blood and plasma in environmentally exposed reproductive-age women (n = 63) living in Mexico City, Mexico. We collected whole blood and plasma samples using trace metal clean techniques and analyzed them for Pb using high-resolution inductively coupled plasma mass spectrometry. A subset of subjects provided repeated blood samples weekly for 4 consecutive weeks (n = 17 subjects) or every 1-2 months over a 9-month period (n = 14 subjects). Plasma Pb concentration was significantly positively associated with whole-blood Pb in a curvilinear fashion over the range of blood Pb values observed here (2.13-39.7 microg/dL). This relationship was best described by the function Plasma Pb = e (-2.392 + 0.0898 x blood Pb), where SE(coefficient) = 0.0054, SE(constant) = 0.063 (n = 63 subjects, n = 141 observations). Results from the short- and long-term repeated collection subjects indicated that the within- and between-subject variance components were not significantly different between the two subsets of subjects. The between subjects component accounts for 78% of the variance in plasma Pb levels, while the residual variance (22%) may be attributed to other unmeasured factors. Collectively, this study demonstrates that plasma Pb measurements may be applied to general clinical settings, provided that established trace metal clean techniques are adopted. This study also shows that the relative (%) partitioning of whole-blood Pb in plasma naturally varies by a factor of about 2-4-fold among subjects at a given blood Pb level. Because Pb in the plasma is considered to more closely represent the fraction of Pb in the circulation that is readily exchanged with peripheral target tissues (e.g., brain, kidney, skeleton), the routine assessment of plasma Pb may provide a more meaningful measure of toxicologically available Pb. PMID- 11882479 TI - Octylphenol and UV-B radiation alter larval development and hypothalamic gene expression in the leopard frog (Rana pipiens). AB - We assessed octylphenol (OP), an estrogenic endocrine-disrupting chemical, and UV B radiation, a known stressor in amphibian development, for their effects on hypothalamic gene expression and premetamorphic development in the leopard frog Rana pipiens. Newly hatched tadpoles were exposed for 10 days to OP alone at two different dose levels; to subambient UV-B radiation alone; and to two combinations of OP and UV-B. Control animals were exposed to ethanol vehicle (0.01%) exposure, a subset of tadpoles from each treatment group was raised to metamorphosis to assess differences in body weight and time required for hindlimb emergence. Tadpoles from one of the OP/UV-B combination groups had greater body weight and earlier hindlimb emergence (p < 0.05), but neither OP nor UV-B alone produced significant changes in body weight or hindlimb emergence, indicating a potential mechanism of interaction between OP and UV-B. We hypothesized that the developing hypothalamus might be a potential environmental sensor for neurotoxicologic studies because of its role in the endocrine control of metamorphosis. We used a differential display strategy to identify candidate genes differentially expressed in the hypothalamic region of the exposed tadpoles. Homology cloning was performed to obtain R. pipiens glutamate decarboxylases--GAD65 and GAD67, enzymes involved in the synthesis of the neurotransmitter gamma-aminobutyric acid (GABA). cDNA expression profiles revealed that OP and UV-B affected the levels of several candidate transcripts in tadpole (i.e., Nck, Ash, and phospholipase C gamma-binding protein 4 and brain angiogenesis inhibitor-3) and metamorph (i.e., GAD67, cytochrome C oxidase, and brain angiogenesis inhibitor-2 and -3) brains. This study represents a novel approach in toxicology that combines physiologic and molecular end points and indicates that levels of OP commonly found in the environment and subambient levels of UV-B alter the expression of important hypothalamic genes and disrupt tadpole growth patterns. PMID- 11882478 TI - Octylphenol (OP) alters the expression of members of the amyloid protein family in the hypothalamus of the snapping turtle, Chelydra serpentina serpentina. AB - The gonadal estrogen estradiol-17beta (E(2)) is important for developing and regulating hypothalamic function and many aspects of reproduction in vertebrates. Pollutants such as octylphenol (OP) that mimic the actions of estrogens are therefore candidate endocrine-disrupting chemicals. We used a differential display strategy (RNA-arbitrarily primed polymerase chain reaction) to isolate partial cDNA sequences of neurotransmitter, developmental, and disease-related genes that may be regulated by OP or E(2) in the snapping turtle Chelydra serpentina serpentina hypothalamus. Hatchling and year-old male snapping turtles were exposed to a 10 ng/mL nominal concentration of waterborne OP or E(2) for 17 days. One transcript [421 base pairs (bp)] regulated by OP and E(2) was 93% identical to human APLP-2. APLP-2 and the amyloid precursor protein (APP) regulate neuronal differentiation and are also implicated in the genesis of Alzheimer disease in humans. Northern blot analysis determined that the turtle hypothalamus contains a single APLP-2 transcript of 3.75 kb in length. Exposure to OP upregulated hypothalamic APLP-2 mRNA levels 2-fold (p < 0.05) in month-old and yearling turtles. E(2) did not affect APLP-2 mRNA levels in hatchlings but stimulated a 2-fold increase (p < 0.05) in APLP-2 mRNA levels in yearling males. The protein beta-amyloid, a selectively processed peptide derived from APP, is also involved in neuronal differentiation, and accumulation of this neurotoxic peptide causes neuronal degeneration in the brains of patients with Alzheimer disease. Therefore, we also sought to determine the effects of estrogens on the expression of beta-amyloid. Using homology cloning based on known sequences, we isolated a cDNA fragment (474 bp) from turtle brain with 88% identity to human APP. Northern blot analysis determined that a single 3.5-kb transcript was expressed in the turtle hypothalamus. Waterborne OP also increased the expression of hypothalamic APP after 35 days of exposure. Our results indicate that low levels of OP are bioactive and can alter the expression of APLP-2 and APP. Because members of the APP gene family are involved in neuronal development, we hypothesize that OP exposure may disrupt hypothalamic development in young turtles. PMID- 11882480 TI - Cancer in wildlife, a case study: beluga from the St. Lawrence estuary, Quebec, Canada. AB - A population of approximately 650 beluga (Delphinapterus leucas) inhabits a short segment of the St. Lawrence estuary (SLE). Over 17 years (1983-1999), we have examined 129 (or 49%) of 263 SLE beluga carcasses reported stranded. The major primary causes of death were respiratory and gastrointestinal infections with metazoan parasites (22%), cancer (18%), and bacterial, viral, and protozoan infections (17%). We observed cancer in 27% of examined adult animals found dead, a percentage similar to that found in humans. The estimated annual rate (AR) of all cancer types (163/100,000 animals) is much higher than that reported for any other population of cetacean and is similar to that of humans and to that of hospitalized cats and cattle. The AR of cancer of the proximal intestine, a minimum figure of 63 per 100,000 animals, is much higher than that observed in domestic animals and humans, except in sheep in certain parts of the world, where environmental contaminants are believed to be involved in the etiology of this condition. SLE beluga and their environment are contaminated by polycyclic aromatic hydrocarbons (PAHs) produced by the local aluminum smelters. The human population living in proximity of the SLE beluga habitat is affected by rates of cancer higher than those found in people in the rest of Quebec and Canada, and some of these cancers have been epidemiologically related to PAHs. Considered with the above observations, the exposure of SLE beluga to PAHs and their contamination by these compounds are consistent with the hypothesis that PAHs are involved in the etiology of cancer in these animals. PMID- 11882481 TI - Effect of motor vehicle emissions on respiratory health in an urban area. AB - Motor vehicles emit particulate matter < 2.5 microm in diameter (PM(2.5)), and as a result, PM(2.5) concentrations tend to be elevated near busy streets. Studies of the relationship between motor vehicle emissions and respiratory health are generally limited by difficulties in exposure assessment. We developed a refined exposure model and implemented it using a geographic information system to estimate the average daily census enumeration area (EA) exposure to PM(2.5). Southeast Toronto, the study area, includes 334 EAs and covers 16 km(2) of urban area. We used hospital admission diagnostic codes from 1990 to 1992 to measure respiratory and genitourinary conditions. We assessed the effect of EA exposure on hospital admissions using a Poisson mixed-effects model and examined the spatial distributions of variables. Exposure to PM(2.5) has a significant effect on admission rates for a subset of respiratory diagnoses (asthma, bronchitis, chronic obstructive pulmonary disease, pneumonia, upper respiratory tract infection), with a relative risk of 1.24 (95% confidence interval, 1.05-1.45) for a log(10) increase in exposure. We noted a weaker effect of exposure on hospitalization for all respiratory conditions, and no effect on hospitalization for nonrespiratory conditions. PMID- 11882482 TI - Mitochondrial membrane potential: a novel biomarker of oxidative environmental stress. AB - Epidemiologic analyses, traditionally based on long-term cohort or case-control studies, provide retrospective causal associations between exposure to a particular environmental stressor and an exposure-related disease end point. Recent research initiatives have propelled a shift toward exploring molecular epidemiology and molecular biological markers (biomarkers) as a means of providing more immediate, quantitative risk assessment of potentially deleterious environmental exposures. We compared, in normal human monocytes isolated from the blood of healthy donors, variations in Hsp70 expression and mitochondrial membrane potential (delta psi m) in response to exposure to either tobacco smoke or gamma-irradiation, two models for environmentally mediated oxidant exposure. On the basis of its mechanistic specificity for oxidants and little baseline variation in cells from distinct individuals, we propose that delta psi m represents a selective in vitro and in vivo biomarker for oxidant exposure. delta psi m may be used to gauge risks associated with oxidant-mediated air pollution and radiation. PMID- 11882483 TI - The association between noise exposure and blood pressure and ischemic heart disease: a meta-analysis. AB - It has been suggested that noise exposure is associated with blood pressure changes and ischemic heart disease risk, but epidemiologic evidence is still limited. Furthermore, most reviews investigating these relations were not carried out in a systematic way, which makes them more prone to bias. We conducted a meta analysis of 43 epidemiologic studies published between 1970 and 1999 that investigate the relation between noise exposure (both occupational and community) and blood pressure and/or ischemic heart disease (International Classification of Diseases, Ninth Revision, codes 410-414). We studied a wide range of effects, from blood pressure changes to a myocardial infarction. With respect to the association between noise exposure and blood pressure, small blood pressure differences were evident. Our meta-analysis showed a significant association for both occupational noise exposure and air traffic noise exposure and hypertension: We estimated relative risks per 5 dB(A) noise increase of 1.14 (1.01-1.29) and 1.26 (1.14-1.39), respectively. Air traffic noise exposure was positively associated with the consultation of a general practitioner or specialist, the use of cardiovascular medicines, and angina pectoris. In cross-sectional studies, road traffic noise exposure increases the risk of myocardial infarction and total ischemic heart disease. Although we can conclude that noise exposure can contribute to the prevalence of cardiovascular disease, the evidence for a relation between noise exposure and ischemic heart disease is still inconclusive because of the limitations in exposure characterization, adjustment for important confounders, and the occurrence of publication bias. PMID- 11882486 TI - Visual contrast sensitivity as a diagnostic tool. PMID- 11882484 TI - Childhood cancer and agricultural pesticide use: an ecologic study in California. AB - We analyzed population-based childhood cancer incidence rates throughout California in relation to agricultural pesticide use. During 1988-1994, a total of 7,143 cases of invasive cancer were diagnosed among children under 15 years of age in California. Building on the availability of high-quality population-based cancer incidence information from the California Cancer Registry, population data from the U.S. Census, and uniquely comprehensive agricultural pesticide use information from California's Department of Pesticide Regulation, we used a geographic information system to assign summary population, exposure, and outcome attributes at the block group level. We used Poisson regression to estimate rate ratios (RRs) by pesticide use density adjusted for race/ethnicity, age, and sex for all types of childhood cancer combined and separately for the leukemias and central nervous system cancers. We generally found no association between pesticide use density and childhood cancer incidence rates. The RR for all cancers was 0.95 [95% confidence interval (CI), 0.80-1.13] for block groups in the 90th percentile and above for use of pesticides classified as probable carcinogens, compared to the block groups with use of < 1 lb/mi(2). The RRs were similar for leukemia and central nervous system cancers. Childhood leukemia rates were significantly elevated (RR = 1.48; 95% CI, 1.03-2.13) in block groups with the highest use of propargite, although we saw no dose-response trend with increasing exposure categories. Results were unchanged by further adjustment for socioeconomic status and urbanization. PMID- 11882487 TI - Men at work--sperm on break. PMID- 11882488 TI - Allergies a la carte: is there a problem with genetically modified foods? PMID- 11882489 TI - Under construction: building a safer industry. AB - A revolution in the building industry over the past decade has spawned a new generation of safer materials and practices, decreasing some health risks for construction workers. Concerned consumers, builders, materials manufacturers, and government regulatory agencies have all contributed to a turn toward "green" building materials and practices, meaning that homeowners and office workers now are better able to live and work in healthier environments, and many construction workers are handling and installing less-toxic materials. PMID- 11882490 TI - Sustainable business makes dollars and sense. AB - The last decade has marked a radical change in the business of sustainability, say environmental activists and industrialists alike. On the wane are "greenwashing" campaigns in which corporations promote dubious environmental accomplishments to draw attention away from environmentally damaging practices. On the rise are corporate investments in sustainability programs, new types of environmental markets, and public demand for green products and investments. Once an afterthought, an annoyance, or a nonentity, sustainability is now often a focal point for businesses. No longer thought of as a business cost, environmentally benign practices are now viewed as a competitive advantage as companies seek to win both stockholders and customers. PMID- 11882491 TI - The hottest thing in remediation. AB - Scientists and engineers are exploring a new way to decontaminate toxic waste sites by literally turning up the heat on pollutants. The method heats the ground using electricity or steam, which mobilizes the contaminants so they can either be extracted from the ground and destroyed or actually destroyed in place. Among the targets for this method are pollutants such as creosote, solvents, and gasoline. These in situ thermal technologies also offer the benefit of reaching contaminants not previously amenable to cleanup, such as those found beneath structures and below the water table. PMID- 11882492 TI - Control of ubiquitination of proteins in rat tissues by ubiquitin conjugating enzymes and isopeptidases. AB - The activity of the ubiquitin-dependent proteolytic system in differentiated tissues under basal conditions remains poorly explored. We measured rates of ubiquitination in rat tissue extracts. Accumulation of ubiquitinated proteins increased in the presence of ubiquitin aldehyde, indicating that deubiquitinating enzymes can regulate ubiquitination. Rates of ubiquitination varied fourfold, with the highest rate in the testis. We tested whether ubiquitin-activating enzyme (E1) or ubiquitin-conjugating enzymes (E2s) could be limiting for conjugation. Immunodepletion of the E2s UBC2 or UBC4 lowered rates of conjugation similarly. Supplementation of extracts with excess UBC2 or UBC4, but not E1, stimulated conjugation. However, UBC2-stimulated rates of ubiquitination still differed among tissues, indicating that tissue differences in E3s or substrate availability may also be rate controlling. UBC2 and UBC4 stimulated conjugation half-maximally at concentrations of 10-50 and 28-44 nM, respectively. Endogenous tissue levels of UBC2, but not UBC4, appeared saturating for conjugation, suggesting that in vivo modulation of UBC4 levels can likely control ubiquitin conjugation. Thus the pool of ubiquitin conjugates and therefore the rate of degradation of proteins by this system may be controlled by E2s, E3s, and isopeptidases. The regulation of the ubiquitin pathway appears complex, but precise. PMID- 11882493 TI - High glucose abolishes the antiproliferative effect of 17beta-estradiol in human vascular smooth muscle cells. AB - We examined effects of 17beta-estradiol (E(2)) on human vascular smooth muscle cell (VSMC) proliferation under normal (5 mmol/l) and high (25 mmol/l) glucose concentrations. Platelet-derived growth factor (PDGF) BB (20 ng/ml)-induced increases in DNA synthesis and proliferation were greater in high than normal glucose concentrations; the difference in DNA synthesis was abolished by a protein kinase C (PKC)-beta inhibitor, LY-379196 (30 nmol/l). Western blotting showed that PKC-beta(1) protein increased in cells exposed to high glucose, whereas PKC-alpha protein and total PKC activity remained unchanged, compared with normal glucose cultures. In normal glucose, E(2) (1-100 nmol/l) inhibited PDGF-induced DNA synthesis by 18-37% and cell proliferation by 16-22% in a concentration-dependent manner. The effects of E(2) were blocked by the estrogen receptor (ER) antagonist ICI-182780, indicating ER dependence. In high glucose, the inhibitory effect of E(2) on VSMC proliferation was abolished but was restored in the presence of the PKC-beta inhibitor LY-379196. Thus high glucose enhances human VSMC proliferation and attenuates the antiproliferative effect of E(2) in VSMC via activation of PKC-beta. PMID- 11882495 TI - Proteolysis-inducing factor differentially influences transcriptional regulation in endothelial subtypes. AB - Proteolysis-inducing factor (PIF) is a novel sulfated glycoprotein initially identified as a protein capable of triggering muscle proteolysis during the process of cancer cachexia. Only skeletal muscle and liver exhibit substantial binding of PIF in adult tissue. Here, we demonstrate that PIF induces transcriptional regulation in both the liver endothelial cell line SK-HEP-1 and in human umbilical vein endothelial cells (HUVECs) but not in pulmonary artery endothelial cells. PIF differentially induces activation of nuclear factor kappaB, resulting in the induction of proinflammatory cytokines [interleukin (IL) 8 and IL-6] and increased expression of the cell surface proteins intercellular adhesion molecule-1 and vascular cell adhesion molecule in SK-HEP-1 and HUVECs only. In addition, PIF induces the shedding of syndecans from the cell surface. Syndecans are involved in wound repair, metastasis of cancers, and embryonic development. These results suggest that PIF may play additional roles in the proinflammatory response observed in cancer cachexia but may also have a role without the cachectic process. PMID- 11882494 TI - No effect of menstrual cycle phase on glucose kinetics and fuel oxidation during moderate-intensity exercise. AB - Resting and exercise fuel metabolism was assessed in three different phases of the menstrual cycle, characterized by different levels of estrogen relative to progesterone: early follicular (EF, low estrogen and progesterone), midfollicular (MF, elevated estrogen, low progesterone), and midluteal (ML, elevated estrogen and progesterone). It was hypothesized that exercise glucose utilization and whole body carbohydrate oxidation would decrease sequentially from the EF to the MF to the ML phase. Normal-weight healthy females, experiencing a regular menstrual cycle, were recruited. Subjects were moderately active but not highly trained. Testing occurred after 3 days of diet control and after an overnight fast (12-13 h). Resting (2 h) and exercise (50% maximal O(2) uptake, 90 min) measurements of whole body substrate oxidation, tracer-determined glucose flux, and substrate and hormone concentrations were made. No significant difference was observed in whole body fuel oxidation during exercise in the three phases (nonprotein respiratory exchange ratio: EF 0.84 +/- 0.01, MF 0.85 +/- 0.01, ML 0.85 +/- 0.01) or in rates of glucose appearance or disappearance. There were, however, significantly higher glucose (P < 0.05) and insulin (P < 0.001) concentrations during the first 45 min of exercise in the ML phase vs. EF and MF phases. In conclusion, whole body substrate oxidation and glucose utilization did not vary significantly across the menstrual cycle in moderately active women, either at rest or during 90 min of moderate-intensity exercise. During the ML phase, however, this similar pattern of substrate utilization was associated with greater glucose and insulin concentrations. Both estrogen and progesterone are elevated during the ML phase of the menstrual cycle, suggesting that one or both of these sex steroids may play a role in this response. PMID- 11882496 TI - Elevated endogenous cortisol reduces autonomic neuroendocrine and symptom responses to subsequent hypoglycemia. AB - We tested the hypothesis that increased endogenous cortisol secretion reduces autonomic neuroendocrine and neurogenic symptom responses to subsequent hypoglycemia. Twelve healthy young adults were studied on two separate occasions, once after infusions of a pharmacological dose of alpha-(1-24)-ACTH (100 microg/h) from 0930 to 1200 and 1330 to 1600, which raised plasma cortisol levels to approximately 45 microg/dl on day 1, and once after saline infusions on day 1. Hyperinsulinemic (2.0 mU x kg(-1) x min(-1)) stepped hypoglycemic clamps (90, 75, 65, 55, and 45 mg/dl glucose steps) were performed on the morning of day 2 on both occasions. These markedly elevated antecedent endogenous cortisol levels reduced the adrenomedullary (P = 0.004, final plasma epinephrine levels of 489 +/ 64 vs. 816 +/-113 pg/ml), sympathetic neural (P = 0.0022, final plasma norepinephrine levels of 244 +/-15 vs. 342 +/-22 pg/ml), parasympathetic neural (P = 0.0434, final plasma pancreatic polypeptide levels of 312 +/- 37 vs. 424 +/- 56 pg/ml), and neurogenic (autonomic) symptom (P = 0.0097, final symptom score of 7.1 +/-1.5 vs. 10.6 +/- 1.6) responses to subsequent hypoglycemia. Growth hormone, but not glucagon or cortisol, responses were also reduced. The findings that increased endogenous cortisol secretion reduces autonomic neuroendocrine and neurogenic symptom responses to subsequent hypoglycemia are potentially relevant to cortisol mediation of hypoglycemia-associated autonomic failure, and thus a vicious cycle of recurrent iatrogenic hypoglycemia, in people with diabetes mellitus. PMID- 11882498 TI - Middle-aged C57BL/6 mice have impaired responses to leptin that are not improved by calorie restriction. AB - Midlife weight gain occurs in many species, suggesting that leptin signaling is impaired at middle age. To test this hypothesis, we measured changes in food intake and body composition in young (Y) and middle-aged (MA) C57BL/6 male mice infused subcutaneously with phosphate-buffered saline or leptin. Leptin-induced decreases in food intake and body fat were delayed in MA mice and associated with catabolism after longer treatment periods. Endogenous plasma leptin levels did not correlate with body fat in MA mice. Calorie restriction (CR) reduced body fat, plasma leptin, and insulin in MA mice to levels in Y mice but did not upregulate leptin sensitivity. CR mice did not respond to leptin doses that inhibited food intake in MA mice and reduced food intake and body fat in Y mice significantly below levels in CR mice. Plasma corticosterone was significantly higher in leptin-treated CR vs. MA mice. We conclude that MA C57BL/6 mice exhibit impaired leptin signaling and that CR, possibly by elevating glucocorticoids, impairs appetite control without improving the metabolic actions of leptin. PMID- 11882497 TI - Reduced activity of mtTFA decreases the transcription in mitochondria isolated from diabetic rat heart. AB - To evaluate abnormalities in the mitochondrial transcription factor A (mtTFA) function as a cause of mitochondrial dysfunction in diabetes, we measured the mRNA contents of the proteins consisting of the mitochondrial respiratory chain as well as transcriptional and translational activities in the mitochondria isolated from controls and streptozotocin-induced diabetic rat hearts. Using Northern blot analysis, we found 40% reduced mRNA contents of mitochondrial encoded cytochrome b and ATP synthase subunit 6 in diabetic rat hearts compared with control rats (P < 0.05). These abnormalities were completely recovered by insulin treatment. Furthermore, the mitochondrial activities of transcription and translation were decreased significantly in mitochondria isolated from diabetic rats by 60% (P < 0.01) and 71% (P < 0.01), respectively, compared with control rats. The insulin treatment also completely normalized these abnormalities in diabetic rats. Consistently, gel retardation assay showed a reduced binding of mtTFA to the D-loop of mitochondrial DNA in diabetic rats, although there was no difference in the mtTFA mRNA and protein content between the two groups. On the basis of these findings, a reduced binding activity of mtTFA to the D-loop region in the hearts of diabetic rats may contribute to the decreased mitochondrial protein synthesis. PMID- 11882499 TI - Glucose release from GLUT2-null hepatocytes: characterization of a major and a minor pathway. AB - We previously reported that glucose can be released from GLUT2-null hepatocytes through a membrane traffic-based pathway issued from the endoplasmic reticulum. Here, we further characterized this glucose release mechanism using biosynthetic labeling protocols. In continuous pulse-labeling experiments, we determined that glucose secretion proceeded linearly and with the same kinetics in control and GLUT2-null hepatocytes. In GLUT2-deficient hepatocytes, however, a fraction of newly synthesized glucose accumulated intracellularly. The linear accumulation of glucose in the medium was inhibited in mutant, but not in control, hepatocytes by progesterone and low temperature, as previously reported, but, importantly, also by microtubule disruption. The intracellular pool of glucose was shown to be present in the cytosol, and, in pulse-chase experiments, it was shown to be released at a relatively slow rate. Release was not inhibited by S-4048 (an inhibitor of glucose-6-phosphate translocase), cytochalasin B, or progesterone. It was inhibited by phloretin, carbonyl cyanide p (trifluoromethoxy)phenylhydrazone, and low temperature. We conclude that the major release pathway segregates glucose away from the cytosol by use of a membrane traffic-based, microtubule-dependent mechanism and that the release of the cytosolic pool of newly synthesized glucose, through an as yet unidentified plasma membrane transport system, cannot account for the bulk of glucose release. PMID- 11882500 TI - Mitochondrial biogenesis during skeletal muscle regeneration. AB - Myogenesis requires energy production for the execution of a number of regulatory and biosynthesis events. We hypothesized that mitochondrial biogenesis would be stimulated during skeletal muscle regeneration. Tibialis anterior muscles of male Sprague-Dawley rats were injected with 0.75% bupivacaine and removed at 3, 5, 7, 10, 14, 21, or 35 days after injection (n = 5-7/group). Two main periods emerged from the histochemical analyses of muscle sections and the expression of proliferating cell nuclear antigen, desmin, and creatine phosphokinase: 1) activation/proliferation of satellite cells (days 3-14) and 2) differentiation into muscle fibers (days 5-35). The onset of muscle differentiation was accompanied by a marked stimulation of mitochondrial biogenesis, as indicated by a nearly fivefold increase in citrate synthase activity and state 3 rate of respiration between days 5 and 10. Peroxisome proliferator-activated receptor gamma coactivator-1 (PGC-1) mRNA level and mitochondrial transcription factor A (mtTFA) protein level peaked on day 10 concurrently with the state 3 rate of respiration. Therefore, transcriptional activation by PGC-1 and mtTFA may be one of the mechanisms regulating mitochondrial biogenesis in regenerating skeletal muscle. Taken together, our results suggest that mitochondrial biogenesis may be an important regulatory event during muscle regeneration. PMID- 11882501 TI - Function-specific calcium stores selectively regulate growth hormone secretion, storage, and mRNA level. AB - Ca(+) stores may regulate multiple components of the secretory pathway. We examined the roles of biochemically independent intracellular Ca(2+) stores on acute and long-term growth hormone (GH) release, storage, and mRNA levels in goldfish somatotropes. Thapsigargin-evoked intracellular Ca(2+) concentration ([Ca(2+)](i)) signal amplitude was similar to the Ca(2+)-mobilizing agonist gonadotropin-releasing hormone, but thapsigargin (2 microM) did not acutely increase GH release, suggesting uncoupling between [Ca(2+)](i) and exocytosis. However, 2 microM thapsigargin affected long-term secretory function. Thapsigargin-treated cells displayed a steady secretion of GH (2, 12, and 24 h), which decreased GH content (12 and 24 h), but not GH mRNA/production (24 h). In contrast to the results with thapsigargin, activating the ryanodine (Ry) receptor (RyR) with 1 nM Ry transiently increased GH release (2 h). Prolonged activation of RyR (24 h) reduced GH release, contents and apparent production, without changing GH mRNA levels. Inhibiting RyR with 10 microM Ry increased GH mRNA levels, production, and storage (2 h). Increasing [Ca(2+)](i) independently of Ca(2+) stores with the use of 30 mM KCl decreased GH mRNA. Collectively, these results suggest that parts of the secretory pathway can be controlled independently by function-specific Ca(2+) stores. PMID- 11882502 TI - Stimulation of insulin secretion and associated nuclear accumulation of iPLA(2)beta in INS-1 insulinoma cells. AB - Accumulating evidence suggests that the cytosolic calcium-independent phospholipase A(2) (iPLA(2)beta) manifests a signaling role in insulin-secreting (INS-1) beta-cells. Earlier, we reported that insulin-secretory responses to cAMP elevating agents are amplified in iPLA(2)beta-overexpressing INS-1 cells (Ma Z, Ramanadham S, Bohrer A, Wohltmann M, Zhang S, and Turk J. J Biol Chem 276: 13198 13208, 2001). Here, immunofluorescence, immunoaffinity, and enzymatic activity analyses are used to examine distribution of iPLA(2)beta in stimulated INS-1 cells in greater detail. Overexpression of iPLA(2)beta in INS-1 cells leads to increased accumulation of iPLA(2)beta in the nuclear fraction. Increasing glucose concentrations alone results in modest increases in insulin secretion, relative to parental cells, and in nuclear accumulation of the iPLA(2)beta protein. In contrast, cAMP-elevating agents induce robust increases in insulin secretion and in time-dependent nuclear accumulation of iPLA(2)beta fluorescence, which is reflected by increases in nuclear iPLA(2)beta protein content and specific enzymatic activity. The stimulated effects are significantly attenuated in the presence of cell-permeable inhibitors of protein phosphorylation and glycosylation. These findings suggest that conditions that amplify insulin secretion promote translocation of beta-cell iPLA(2)beta to the nuclei, where it may serve a crucial signaling role. PMID- 11882503 TI - Heterogeneous metabolic adaptation of C57BL/6J mice to high-fat diet. AB - C57BL/6J mice were fed a high-fat, carbohydrate-free diet (HFD) for 9 mo. Approximately 50% of the mice became obese and diabetic (ObD), approximately 10% lean and diabetic (LD), approximately 10% lean and nondiabetic (LnD), and approximately 30% displayed intermediate phenotype. All of the HFD mice were insulin resistant. In the fasted state, whole body glucose clearance was reduced in ObD mice, unchanged in the LD mice, and increased in the LnD mice compared with the normal-chow mice. Because fasted ObD mice were hyperinsulinemic and the lean mice slightly insulinopenic, there was no correlation between insulin levels and increased glucose utilization. In vivo, tissue glucose uptake assessed by 2 [(14)C]deoxyglucose accumulation was reduced in most muscles in the ObD mice but increased in the LnD mice compared with the values of the control mice. In the LD mice, the glucose uptake rates were reduced in extensor digitorum longus (EDL) and total hindlimb but increased in soleus, diaphragm, and heart. When assessed in vitro, glucose utilization rates in the absence and presence of insulin were similar in diaphragm, soleus, and EDL muscles isolated from all groups of mice. Thus, in genetically homogenous mice, HFD feeding lead to different metabolic adaptations. Whereas all of the mice became insulin resistant, this was associated, in obese mice, with decreased glucose clearance and hyperinsulinemia and, in lean mice, with increased glucose clearance in the presence of mild insulinopenia. Therefore, increased glucose clearance in lean mice could not be explained by increased insulin level, indicating that other in vivo mechanisms are triggered to control muscle glucose utilization. These adaptive mechanisms could participate in the protection against development of obesity. PMID- 11882504 TI - Pronatriuretic peptide is a sensitive marker of the endocrine function of teleost heart. AB - We recently characterized a novel heart-specific hormone from salmon (salmon cardiac peptide, sCP). We have now prepared a recombinant plasmid expressing the NH(2)-terminal fragment of pro-sCP (NT-pro-sCP) and used it to set up a specific RIA for the peptide. Because of the sensitivity of the assay and the high circulating levels, NT-pro-sCP can be measured from as little as 2 microl of serum. This enables repeated sampling from the same animal in different experimental setups. Mechanical load increased the release of NT-pro-sCP from isolated perfused salmon ventricle, in parallel with sCP. Bolus injection of human endothelin-1 (ET-1; 1 microg) in the dorsal aorta of salmon resulted in an extensive increase of serum NT-pro-sCP (from 0.99 +/- 0.11 to 4.6 +/-1.5 nmol/l). The response was abolished by pretreatment with a specific type A ET (ET(A)) receptor antagonist (BQ-123) but not with a type B ET receptor antagonist (BQ 788). The NT-pro-sCP levels had a good correlation with those of sCP (r(2) = 0.75). Our results demonstrate the practical usefulness of circulating NT-pro-sCP as a marker of the endocrine function of salmon heart. They also suggest that ET 1 has an important role in regulating sCP release from teleost heart by an ET(A) receptor-mediated mechanism. PMID- 11882505 TI - Bone as an ion exchange system: evidence for a link between mechanotransduction and metabolic needs. AB - To detect whether the mutual interaction occurring between the osteocytes-bone lining cells system (OBLCS) and the bone extracellular fluid (BECF) is affected by load through a modification of the BECF-extracellular fluid (ECF; systemic extracellular fluid) gradient, mice metatarsal bones immersed in ECF were subjected ex vivo to a 2-min cyclic axial load of different amplitudes and frequencies. The electric (ionic) currents at the bone surface were measured by a vibrating probe after having exposed BECF to ECF through a transcortical hole. The application of different loads and different frequencies increased the ionic current in a dose-dependent manner. The postload current density subsequently decayed following an exponential pattern. Postload increment's amplitude and decay were dependent on bone viability. Dummy and static loads did not induce current density modifications. Because BECF is perturbed by loading, it is conceivable that OBLCS tends to restore BECF preload conditions by controlling ion fluxes at the bone-plasma interface to fulfill metabolic needs. Because the electric current reflects the integrated activity of OBLCS, its evaluation in transgenic mice engineered to possess genetic lesions in channels or matrix constituents could be helpful in the characterization of the mechanical and metabolic functions of bone. PMID- 11882506 TI - Increase in daily LH secretion in response to short-term calorie restriction in obese women with PCOS. AB - We hypothesized that short-term calorie restriction would blunt luteinizing hormone (LH) hypersecretion in obese women with polycystic ovary syndrome (PCOS) and thereby ameliorate the anovulatory endocrine milieu. To test this hypothesis, 15 obese patients with PCOS and nine age- and body mass index-matched healthy women underwent 24-h blood sampling to quantitate plasma LH, leptin, and insulin levels. PCOS subjects were prescribed a very low caloric liquid diet (4.2 MJ/day) for 7 days and were then resampled. Basal and pulsatile LH secretion was threefold higher in PCOS subjects, but plasma insulin and leptin levels were not different in the calorie-replete state. Contrary to expectation, calorie restriction enhanced basal and pulsatile LH secretion even further. As expected, plasma glucose, insulin, and leptin concentrations decreased by 18, 75, and 50%, respectively. Serum total testosterone concentration fell by 23%, whereas serum estrone, estradiol, sex hormone-binding globulin (SHBG), and androstenedione concentrations remained unchanged. Enhanced LH secretion in the presence of normal metabolic and hormonal adaptations to calorie restriction points to anomalous feedback control of pituitary LH release in PCOS. PMID- 11882507 TI - GLP-1-(9-36) amide reduces blood glucose in anesthetized pigs by a mechanism that does not involve insulin secretion. AB - Glucagon-like peptide 1 (GLP-1) is a potent anti-hyperglycemic hormone currently under investigation for its therapeutic potential. However, due to rapid degradation by dipeptidyl peptidase IV (DPP IV), which limits its metabolic stability and eliminates its insulinotropic activity, it has been impossible to assess its true efficacy in vivo. In chloralose-anesthetized pigs given valine pyrrolidide (to block endogenous DPP IV activity), the independent effects of GLP 1-(7-36) amide on glucose and insulin responses to intravenous glucose were assessed, and the metabolite generated by DPP IV, GLP-1-(9-36) amide, was investigated for any ability to influence these responses. GLP-1-(7-36) amide enhanced insulin secretion (P < 0.03 vs. vehicle), but GLP-1-(9-36) amide was without effect, either alone or when coinfused with GLP-1-(7-36) amide. In contrast, GLP-1-(9-36) amide did affect glucose responses (P < 0.03). Glucose excursions were greater after saline (121 +/- 17 mmol x l(-1) x min) than after GLP-1-(9-36) amide (73 +/- 19 mmol x l(-1) x min; P < 0.05), GLP-1-(7-36) amide (62 +/- 13 mmol x l(-1) x min; P < 0.02) or GLP-1-(7-36) amide + GLP-1-(9-36) amide (50 +/-13 mmol x l(-1) x min; P < 0.005). Glucose elimination rates were faster after GLP-1-(7-36) amide + (9-36) amide (10.3 +/- 1.2%/min) than after GLP 1-(7-36) amide (7.0 +/- 0.9%/min; P < 0.04), GLP-1-(9-36) amide (6.8 +/- 1.0%/min; P < 0.03), or saline (5.4 +/- 1.2%/min; P < 0.005). Glucagon concentrations were unaffected. These results demonstrate that GLP-1-(9-36) amide neither stimulates insulin secretion nor antagonizes the insulinotropic effect of GLP-1-(7-36) amide in vivo. Moreover, the metabolite itself possesses anti hyperglycemic effects, supporting the hypothesis that selective DPP IV action is important in glucose homeostasis. PMID- 11882508 TI - Stimulation of protein synthesis by both insulin and amino acids is unique to skeletal muscle in neonatal pigs. AB - In neonatal pigs, the feeding-induced stimulation of protein synthesis in skeletal muscle, but not liver, can be reproduced by insulin infusion when essential amino acids and glucose are maintained at fasting levels. In the present study, 7- and 26-day-old pigs were studied during 1) fasting, 2) hyperinsulinemic-euglycemic-euaminoacidemic clamps, 3) euinsulinemic-euglycemic hyperaminoacidemic clamps, and 4) hyperinsulinemic-euglycemic-hyperaminoacidemic clamps. Amino acids were clamped using a new amino acid mixture enriched in nonessential amino acids. Tissue protein synthesis was measured using a flooding dose of L-[4-(3)H]phenylalanine. In 7-day-old pigs, insulin infusion alone increased protein synthesis in various skeletal muscles (from +35 to +64%), with equivalent contribution of myofibrillar and sarcoplasmic proteins, as well as cardiac muscle (+50%), skin (+34%), and spleen (+26%). Amino acid infusion alone increased protein synthesis in skeletal muscles (from +28 to +50%), also with equivalent contribution of myofibrillar and sarcoplasmic proteins, as well as liver (+27%), pancreas (+28%), and kidney (+10%). An elevation of both insulin and amino acids did not have an additive effect. Similar qualitative results were obtained in 26-day-old pigs, but the magnitude of the stimulation of protein synthesis by insulin and/or amino acids was lower. The results suggest that, in the neonate, the stimulation of protein synthesis by feeding is mediated by either amino acids or insulin in most tissues; however, the feeding-induced stimulation of protein synthesis in skeletal muscle is uniquely regulated by both insulin and amino acids. PMID- 11882509 TI - Ligand-specific regulation of proteasome-mediated proteolysis of estrogen receptor-alpha. AB - Proteasome-mediated proteolysis modulates the cellular concentration of estrogen receptor-alpha (ERalpha) and is induced by treatment of cells with 17beta estradiol. Herein, we show that multiple receptor agonists, including 17alpha estradiol and estriol as well as the antagonist ICI-182780, stimulate proteasome dependent proteolysis of ERalpha in a process that requires ligand binding to the receptor. Proteolysis of receptor depends on ligand concentration, and there exists a direct correlation between ligand-binding affinity and the half-maximal dose of ligand required to stimulate receptor degradation. Furthermore, introduction of a point mutation into the receptor ligand-binding pocket yields a stable receptor resistant to proteolysis. Interestingly, although all ligands stimulate receptor degradation, the extent to which overall ER levels are affected varies with each ligand and is not related to ligand-binding affinity or activation of transcription. These results demonstrate ligand-specific regulation of ERalpha proteolysis, and they introduce the concept that cellular receptor concentration is governed not only at the level of induction of proteolysis but also by the efficiency with which the receptor is degraded. PMID- 11882510 TI - Increased synthesis rate of fibrinogen as a basis for its elevated plasma levels in obese female adolescents. AB - Increased concentrations of plasma fibrinogen, an independent risk factor for cardiovascular disease (CVD), in obese children have been reported. The underlying mechanism for this, however, remains to be defined. In the current study, we measured the fractional synthesis rates (FSR) of plasma fibrinogen in six healthy postpubertal obese girls [body mass index (BMI) 36.6 +/- 1.8 kg/m(2); age 16.6 +/- 0.5 yr] and six age-matched lean normal control girls (BMI 20.8 +/- 0.7 kg/m(2); age 16.4 +/- 0.4 yr) during a primed, continuous infusion of L-[1 (13)C]leucine in the postabsorptive state. The method involved purification of plasma fibrinogen by use of immunoaffinity chromatography followed by measurement of [(13)C]leucine enrichment using gas chromatography-combustion-isotope ratio mass spectrometry. The FSR of fibrinogen in obese girls (35.06 +/- 2.61%/day) was almost double that in lean girls (17.02 +/- 1.43%/day), and this increase was associated with a relative increase in plasma concentration of fibrinogen as well as BMI in the subjects studied. Obese subjects had high fasting insulin levels (138 +/- 47 pmol/l) compared with lean subjects (54 +/- 11 pmol/l), whereas their glucose concentrations were similar (4.5 +/- 0.3 mmol/l in obese and 4.4 +/- 0.4 mmol/l in lean subjects), suggesting insulin resistance. The doubling of the FSR of fibrinogen provides novel insight into the mechanism of elevated levels of plasma fibrinogen and suggests a primary role for increased synthesis in producing the hyperfibrinogenemia associated with obesity. This finding may have important implications in the design of therapies for modulating plasma fibrinogen levels in obesity and/or CVD in childhood. PMID- 11882511 TI - Activation of liver G-6-Pase in response to insulin-induced hypoglycemia or epinephrine infusion in the rat. AB - This study was conducted to test the hypothesis of the activation of glucose-6 phosphatase (G-6-Pase) in situations where the liver is supposed to sustain high glucose supply, such as during the counterregulatory response to hypoglycemia. Hypoglycemia was induced by insulin infusion in anesthetized rats. Despite hyperinsulinemia, endogenous glucose production (EGP), assessed by [3 (3)H]glucose tracer dilution, was paradoxically not suppressed in hypoglycemic rats. G-6-Pase activity, assayed in a freeze-clamped liver lobe, was increased by 30% in hypoglycemia (P < 0.01 vs. saline-infused controls). Infusion of epinephrine (1 microg x kg(-1) x min(-1)) in normal rats induced a dramatic 80% increase in EGP and a 60% increase in G-6-Pase activity. In contrast, infusion of dexamethasone had no effect on these parameters. Similar insulin-induced hypoglycemia experiments performed in adrenalectomized rats did not induce any stimulation of G-6-Pase. Infusion of epinephrine in adrenalectomized rats restored a stimulation of G-6-Pase similar to that triggered by hypoglycemia in normal rats. These results strongly suggest that specific activatory mechanisms of G-6-Pase take place and contribute to EGP in situations where the latter is supposed to be sustained. PMID- 11882513 TI - Acute hyperglycemia alters the ability of the normal beta-cell to sense and respond to glucose. AB - Impaired glucose tolerance (IGT) and non-insulin-dependent diabetes mellitus (NIDDM) are associated with an impaired ability of the beta-cell to sense and respond to small changes in plasma glucose. The aim of this study was to establish whether acute hyperglycemia per se plays a role in inducing this defect in beta-cell response. Seven healthy volunteers with no family history of NIDDM were studied on two occasions during a 12-h oscillatory glucose infusion with a periodicity of 144 min. Once, low-dose glucose was infused at a mean rate of 6 mg x kg(-1) x min(-1) and amplitude 33% above and below the mean rate, and, once, high-dose glucose was infused at 12 mg x kg(-1) x min(-1) and amplitude 16% above and below the mean rate. Mean glucose levels were significantly higher during the high-dose compared with the low-dose glucose infusion [9.5 +/- 0.8 vs. 6.8 +/- 0.2 mM (P < 0.01)], resulting in increased mean insulin secretion rates [ISRs; 469.1 +/- 43.8 vs. 268.4 +/- 29 pmol/min (P < 0.001)] and mean insulin levels [213.6 +/- 46 vs. 67.9 +/- 10.9 pmol/l (P < 0.008)]. Spectral analysis evaluates the regularity of oscillations in glucose, insulin secretion, and insulin at a predetermined frequency. Spectral power for glucose, ISR, and insulin was reduced during the high-dose glucose infusion [11.8 +/- 1.4 to 7.0 +/- 1.6 (P < 0.02), 7.6 +/- 1.5 to 3.2 +/- 0.5 (P < 0.04), and 10.5 +/- 1.6 to 4.6 +/- 0.7 (P < 0.01), respectively]. In conclusion, short-term infusion of high-dose glucose to obtain glucose levels similar to those previously seen in IGT subjects results in reduced spectral power for glucose, ISR, and insulin. The reduction in spectral power previously observed for ISR in IGT or NIDDM subjects may be due partly to hyperglycemia. PMID- 11882512 TI - Gastrointestinal absorption and plasma kinetics of soy Delta(5)-phytosterols and phytostanols in humans. AB - Our objective was to measure the systemic absorption of lecithin-emulsified Delta(5)-phytosterols and phytostanols during test meals by use of dual stable isotopic tracers. Ten healthy subjects underwent two single-meal absorption tests in random order 2 wk apart, one with intravenous dideuterated Delta(5) phytosterols and oral pentadeuterated Delta(5)-phytosterols and the other with the corresponding labeled stanols. The oral-to-intravenous tracer ratio in plasma, a reflection of absorption, was measured by a sensitive negative ion mass spectroscopic technique and became constant after 2 days. Absorption from 600 mg of Delta(5)-soy sterols given with a standard test breakfast was 0.512 +/- 0.038% for sitosterol and 1.89 +/- 0.27% for campesterol. The absorption from 600 mg of soy stanols was 0.0441 +/- 0.004% for sitostanol and 0.155 +/- 0.017% for campestanol. Reduction of the double bond at position 5 decreased absorption by 90%. Plasma t(1/2) for stanols was significantly shorter than that for Delta(5) sterols. We conclude that the efficiency of phytosterol absorption is lower than what was reported previously and is critically dependent on the structure of both sterol nucleus and side chain. PMID- 11882514 TI - Pattern and cost of weight gain in previously obese women. AB - Weight gain is common among postobese individuals, providing an opportunity to address the cost of weight regain on energy expenditure. We investigated the energy cost of weight regain over 1 yr in 28 women [age 39.5 +/- 1.3 (SE) yr; body mass index 24.2 +/- 0.5 kg/m(2)] with recent weight loss (>12 kg). Body composition, total energy expenditure (TEE) using doubly labeled water, resting metabolic rate (RMR), and thermic effect of a meal (TEM) were assessed at 0 and 12 mo. Metabolizable energy intake (MEI) was calculated from TEE and change in body composition. Fourteen women had a weight gain of 13.2 +/- 2.1 kg. Twelve month cumulative excess MEI, calculated as the intake in excess of TEE at month 0, was 749 +/- 149 MJ. Of this, 462 +/- 83 MJ (62%) were stored as accrued tissue, and 287 +/- 72 MJ (38%) was increased TEE. Expressed per kilogram of body weight gain, the energy cost of weight gain was calculated to be 54.8 +/- 4.6 MJ/kg. Interestingly, weight regain time courses fell into three distinct patterns, possibly requiring varying countermeasures. PMID- 11882515 TI - Regional muscle and adipose tissue amino acid metabolism in lean and obese women. AB - The effect of obesity on regional skeletal muscle and adipose tissue amino acid metabolism is not known. We evaluated systemic and regional (forearm and abdominal subcutaneous adipose tissue) amino acid metabolism, by use of a combination of stable isotope tracer and arteriovenous balance methods, in five lean women [body mass index (BMI) <25 kg/m(2)] and five women with abdominal obesity (BMI 35.0-39.9 kg/m(2); waist circumference >100 cm) who were matched on fat-free mass (FFM). All subjects were studied at 22 h of fasting to ensure that the subjects were in net protein breakdown during this early phase of starvation. Leucine rate of appearance in plasma (an index of whole body proteolysis), expressed per unit of FFM, was not significantly different between lean and obese groups (2.05 +/- 0.18 and 2.34 +/- 0.04 micromol x kg FFM(-1) x min(-1), respectively). However, the rate of leucine release from forearm and adipose tissues in obese women (24.0 +/- 4.8 and 16.6 +/- 6.5 nmol x 100 g(-1) x min(-1), respectively) was lower than in lean women (66.8 +/- 10.6 and 38.6 +/- 7.0 nmol x 100 g(-1) x min(-1), respectively; P < 0.05). Approximately 5-10% of total whole body leucine release into plasma was derived from adipose tissue in lean and obese women. The results of this study demonstrate that the rate of release of amino acids per unit of forearm and adipose tissue at 22 h of fasting is lower in women with abdominal obesity than in lean women, which may help obese women decrease body protein losses during fasting. In addition, adipose tissue is a quantitatively important site for proteolysis in both lean and obese subjects. PMID- 11882516 TI - Correlation between GABA release from rat islet beta-cells and their metabolic state. AB - Pancreatic beta-cells express glutamate decarboxylase (GAD), which is responsible for the production and release of gamma-aminobutyric acid (GABA). Over a 24-h culture period, total GABA release by purified rat beta-cells is eightfold higher than the cellular GABA content and can thus be used as an index of cellular GAD activity. GABA release is 40% reduced by glucose (58 pmol/10(3) cells at 10 mM glucose vs. 94 pmol at 3 mM glucose, P < 0.05). This suppressive effect of glucose was not observed when glucose metabolism was blocked by mannoheptulose or 2,4-dinitrophenol; it was amplified when ATP-dependent beta-cell activities were inhibited by addition of diazoxide, verapamil, or cycloheximide or by reduction of extracellular calcium levels; it was counteracted when beta-cell functions were activated by nonmetabolized agents, such as glibenclamide, IBMX, glucagon, or glucacon-like peptide-1 (GLP-1), which are known to stimulate calcium dependent activities, such as hormone release and calcium-dependent ATPases. These observations suggest that GABA release from beta-cells varies with the balance between ATP-producing and ATP-consuming activities in the cells. Less GABA is released in conditions of elevated glucose metabolism, and hence ATP production, but this effect is counteracted by ATP-dependent activities. The notion that increased cytoplasmic ATP levels can suppress GAD activity in beta cells, and hence GABA production and release, is compatible with previous findings on ATP suppression of brain GAD activity. PMID- 11882517 TI - Pulsatile growth hormone secretion persists in genetic growth hormone-releasing hormone resistance. AB - Growth hormone (GH) secretion is regulated by GH-releasing hormone (GHRH), somatostatin, and possibly ghrelin, but uncertainty remains about the relative contributions of these hypophysiotropic factors to GH pulsatility. Patients with genetic GHRH receptor (GHRH-R) deficiency present an opportunity to examine GH secretory dynamics in the selective absence of GHRH input. We studied circadian GH profiles in four young men homozygous for a null mutation in the GHRH-R gene by use of an ultrasensitive GH assay. Residual GH secretion was pulsatile, with normal pulse frequency, but severely reduced amplitude (<1% normal) and greater than normal process disorder (as assessed by approximate entropy). Nocturnal GH secretion, both basal and pulsatile, was enhanced compared with daytime. We conclude that rhythmic GH secretion persists in an amplitude-miniaturized version in the absence of a GHRH-R signal. The nocturnal enhancement of GH secretion is likely mediated by decreased somatostatin tone. Pulsatility of residual GH secretion may be caused by oscillations in somatostatin and/or ghrelin; it may also reflect intrinsic oscillations in somatotropes. PMID- 11882518 TI - Shedding of TNF-alpha receptors, blood pressure, and insulin sensitivity in type 2 diabetes mellitus. AB - Tumor necrosis factor-alpha (TNF-alpha) is increasingly recognized as a key component in the development of insulin resistance and increased blood pressure. In a sample of 368 individuals, the ratio of soluble TNF-alpha receptors (sTNFR2/sTNFR1) correlated positively with systolic and diastolic blood pressure (P < 0.01). This ratio was significantly greater in type 2 diabetic subjects (DM 2) than in type 1 diabetic patients and was greater than in control nondiabetic subjects (P < 0.00001). The TNF-alpha receptor 1 (TNFR1) density in peripheral blood monocytes was similar in DM-2 patients and in nondiabetic subjects. After phorbol 12-myristate 13-acetate, TNFR1 shedding was significantly decreased in DM 2 compared with control subjects, and it was directly associated with insulin sensitivity (r = 0.54, P = 0.03). Serum sTNFR1 concentration was also linked to the vasodilatory response to glyceryltrinitrate (P = 0.01). Conversely, TNF-alpha receptor 2 shedding was negatively associated with insulin sensitivity (r = 0.54, P = 0.03), whereas shedding of L-selectin showed no significant association. After exercise-induced lowering of blood pressure, a parallel decrease in sTNFR2/sTNFR1 was observed in DM-2 patients. Our findings suggest that insulin resistance and blood pressure are linked to altered shedding of TNF alpha receptors in DM-2. The latter seems reversible and is not genetically determined. PMID- 11882519 TI - New bioimpedance model accurately predicts lower limb muscle volume: validation by magnetic resonance imaging. AB - Conventional bioimpedance analysis (BIA) methods now simplify the representation of lower limb geometry and electrical properties for body composition estimation. In the present study, a three-dimensional model of the lower limb was assembled by segmentation of magnetic resonance cross-sectional images (MRI) for adipose tissue, skeletal muscle, and bone. An electrical network was then associated with this model. BIA and MRI measurements were made in six lean subjects (3 men and 3 women, age 32.2 +/- 6.9 yr). Assuming 0.85 S/m for the longitudinal conductivity of the muscle, the model predicted in the examined subjects an impedance profile that conformed well to the BIA impedance profile; predicted and measured resistances were similar (261.3 +/- 7.7 vs. 249 +/- 9 Omega; P = not significant). The resistance profile provided, through a simpler model, muscle area estimates along the lower limb and total leg muscle volume (mean 4,534 cm(3) for men and 4,071 cm(3) for women) with a mean of the absolute value of relative error with respect to MRI of 6.2 +/- 3.9. The new approach suggests that BIA can reasonably estimate the distribution and volume of muscles in the lower extremities of lean subjects. PMID- 11882520 TI - Hyperleptinemia in A(y)/a mice upregulates arcuate cocaine- and amphetamine regulated transcript expression. AB - The effects of leptin on cocaine- and amphetamine-regulated transcript (CART) and agouti-related protein (AGRP) expression in the hypothalamic arcuate nucleus of obese A(y)/a mice were investigated. CART mRNA expression was upregulated by 41% and AGRP mRNA downregulated by 78% in hyperleptinemic A(y)/a mice relative to levels in lean a/a mice. The mRNA expression of these neuropeptides in either young nonobese A(y)/a mice or rats treated with SHU-9119, a synthetic melanocortin-4 receptor (MC4R) antagonist, did not differ significantly from that in the corresponding controls. After a 72-h fast, which decreased the concentration of serum leptin, CART and AGRP mRNA expression decreased and increased, respectively, in A(y)/a mice. The expression levels of these neuropeptides in leptin-deficient A(y)/a ob/ob double mutants were comparable to those in a/a ob/ob mice. Leptin thus modulates both CART and AGRP mRNA expression in obese A(y)/a mice, whereas leptin signals are blocked at the MCR4R level. Taken together, the present findings indicate that differential expression of these neuropeptides in A(y)/a and ob/ob mice results in dissimilar progression toward obesity. PMID- 11882521 TI - Nomenclature of the GLUT/SLC2A family of sugar/polyol transport facilitators. AB - The recent identification of several additional members of the family of sugar transport facilitators (gene symbol SLC2A, protein symbol GLUT) has created a heterogeneous and, in part, confusing nomenclature. Therefore, this letter provides a summary of the family members and suggests a systematic nomenclature for SLC2A and GLUT symbols. PMID- 11882522 TI - Apolipoprotein E polymorphism and cardiovascular disease: a HuGE review. AB - This review examines the association between the apolipoprotein (apo) var epsilon gene polymorphism (or its protein product (apo E)), metabolic regulation of cholesterol, and cardiovascular disease. The apo var epsilon gene is located at chromosome 19q13.2. Among the variants of this gene, alleles (*) epsilon2, (*) epsilon3, and (*) epsilon4 constitute the common polymorphism found in most populations. Of these variants, apo (*) epsilon3 is the most frequent (>60%) in all populations studied. The polymorphism has functional effects on lipoprotein metabolism mediated through the hepatic binding, uptake, and catabolism of chylomicrons, chylomicron remnants, very low density lipoprotein (VLDL), and high density lipoprotein subspecies. Apo E is the primary ligand for two receptors, the low density lipoprotein (LDL) receptor (also known as the B/E receptor) found on the liver and other tissues and an apo E-specific receptor found on the liver. The coordinate interaction of these lipoprotein complexes with their receptors forms the basis for the metabolic regulation of cholesterol. Allelic variation in apo var epsilon is consistently associated with plasma concentrations of total cholesterol, LDL cholesterol, and apo B (the major protein of LDL, VLDL, and chylomicrons). Apo var epsilon has been studied in disorders associated with elevated cholesterol levels or lipid derangements (i.e., hyperlipoproteinemia type III, coronary heart disease, strokes, peripheral artery disease, and diabetes mellitus). The apo var epsilon genotype yields poor predictive values when screening for clinically defined atherosclerosis despite positive, but modest associations with plaque and coronary heart disease outcomes. In addition to genotype-phenotype associations with vascular disease, the alleles and isoforms of apo var epsilon have been related to dementias, most commonly Alzheimer's disease. PMID- 11882523 TI - Dose-specific meta-analysis and sensitivity analysis of the relation between alcohol consumption and lung cancer risk. AB - Alcohol drinking increases the risk of several types of cancer, but studies of the relation between alcohol and lung cancer risk are complicated by smoking. The authors carried out meta-analyses for four study designs and conducted sensitivity analyses to assess the results. Pooled smoking-unadjusted relative risks (RRs) for brewery workers and alcoholics were 1.17 (95% confidence interval (CI): 0.99, 1.39) and 1.99 (95% CI: 1.66, 2.39), respectively, relative to population rates. For cohort and case-control studies, the authors conducted dose specific meta-analyses for ethanol consumption of 1-499, 500-999, 1,000-1,999, and > or = 2,000 g/month, relative to nondrinking. Smoking-adjusted RRs for ascending dose groups in cohort studies were 0.98 (95% CI: 0.79, 1.21), 0.92 (95% CI: 0.81, 1.04), 1.04 (95% CI: 0.88, 1.22), and 1.53 (95% CI: 1.04, 2.25), respectively. Smoking-adjusted odds ratios for ascending groups in case-control studies were 0.63 (95% CI: 0.51, 0.78), 1.30 (95% CI: 0.98, 1.70), 1.13 (95% CI: 0.46, 2.75), and 1.86 (95% CI: 1.39, 2.49), respectively. Elevated odds ratios were seen for hospital-based case-control studies but not for population-based case-control studies. Sensitivity analyses indicated that smoking explained the elevated RRs in studies of alcoholics and that strong misclassification of smoking status could produce an elevated smoking-adjusted RR in cohort and case control studies. Overall, evidence for a smoking-adjusted association between alcohol and lung cancer risk is limited to very high consumption groups in cohort and hospital-based case-control studies. At lower levels, any associations observed appear to be explained by confounding. PMID- 11882524 TI - Neighborhood environment and loss of physical function in older adults: evidence from the Alameda County Study. AB - Research suggests that neighborhood environment may influence functional health at an older age. This study examined the association between neighborhood problems and incidence of overall and lower-extremity functional loss. A total of 883 participants in the Alameda County Study who were aged 55 years and older and functionally healthy were questioned in 1994 and 1995 as part of an ongoing cohort study. Participants rated the severity of six neighborhood problems: traffic, noise, crime, trash and litter, lighting, and public transportation. Seventeen percent reported multiple neighborhood problems. Functional loss was measured by self-report of severe difficulty with physical tasks (e.g., climbing stairs, lifting 10 pounds (4.54 kg)). After 1 year, 6.1% developed overall functional loss, and 3.9% developed lower-extremity functional loss. Regression models adjusted for demographic, socioeconomic, health, and behavioral risk factors. Compared with those who reported nonproblem neighborhoods, those who reported multiple-problem neighborhoods were at increased risk of overall functional loss (odds ratio = 2.23, 95% confidence interval: 1.08, 4.60) and lower-extremity functional loss (odds ratio = 3.12, 95% confidence interval: 1.15, 8.51). Neighborhood problems associated with the largest increase in risk were excessive noise, inadequate lighting, and heavy traffic. Older people who reported problematic neighborhood environments had a greater risk of functional deterioration over 1 year compared with those in better neighborhoods. PMID- 11882525 TI - Invited commentary: places, people, and health. PMID- 11882526 TI - Socioeconomic status and mortality among the elderly: findings from four US communities. AB - The effect of socioeconomic status (SES) on mortality was examined in the community-dwelling elderly. Data were obtained from four population-based studies that enrolled elderly residents of four US communities (East Boston, Massachusetts; New Haven, Connecticut; east-central Iowa; and the Piedmont region of North Carolina) and followed them for 9 years, starting in 1982 or 1986. Higher SES, whether measured by education, by household income, or by occupational prestige, was generally associated with lower mortality. However, the pattern of findings varied by gender and by community. For men, all three SES indicators were associated with mortality in the majority of cohorts. For women, this was true only for income. SES-mortality associations were attenuated but not eliminated after adjustment for behavior and health status. SES-mortality associations were stronger in New Haven and North Carolina than in East Boston and Iowa. The latter communities are more homogeneous with respect to ethnicity, urbanization, and occupational history than the former. Future research should investigate the relative validity of traditional SES measures for men and women and develop more balanced assessment methods. These findings also suggest that it is important to consider not only individual characteristics but also community attributes that mediate or modify the pathways through which socioeconomic conditions may influence health. PMID- 11882527 TI - Impact of sociodemographic factors, hormone receptor status, and tumor grade on ethnic differences in tumor stage and size for breast cancer in US women. AB - The importance of sociodemographic factors and tumor biomarkers in explaining ethnic differences in tumor stage and size at diagnosis was investigated in over 106,000 female breast cancer patients reported during 1992-1996 from 11 US population-based cancer registries. Japanese and non-Hispanic White women tended to be diagnosed at an earlier stage, with smaller diameter tumors and with a lower tumor grade than women from seven other ethnic groups. Statistical adjustment for individual- and group-level sociodemographic factors produced 50 80% reductions in the odds ratios for distant (vs. localized) stage and larger (vs. <1 cm) tumor size among Black women and Hispanic women. These factors also helped to account for tumor stage and size variation among most other ethnic groups. Consideration of hormone receptor status and tumor grade had little effect on the ethnic patterns. Although small, elevated odds ratios remained for some groups, our results suggest that sociodemographic factors accounted for many of the observed ethnic differences in breast cancer stage and tumor size at the time of diagnosis. Because most of the sociodemographic variables were aggregate measures, it is possible that residual confounding by socioeconomic position could explain the persistence of slightly elevated odds ratios in some ethnic groups. PMID- 11882528 TI - Smoking and risk of acute myeloid leukemia: results from a Los Angeles County case-control study. AB - Acute myelogenous leukemia (AML) is a heterogeneous disease with distinct histologic subtypes likely to have distinct risk factors. The authors examined smoking and the risk of adult AML by French-American-British (FAB) subtype in a Los Angeles County, California, population-based case-control study of 412 cases diagnosed between 1987 and 1994 and 412 matched controls. Consistent with previous studies, smoking was not a substantial risk factor for AML overall (odds ratio (OR) = 1.2, 95% confidence interval (CI): 0.9, 1.6). However, increased risk was observed for FAB subtype M2 (OR = 2.3, 95% CI: 1.1, 4.4), particularly for subjects aged 60-75 years (OR = 3.3, 95% CI: 1.1, 10.0). For M2, significant dose-response was associated with total years smoked (p = 0.02), cigarettes per day (p = 0.007), and product filter status (filtered vs. nonfiltered; p = 0.03). The authors estimate that 42% (standard error = 13%) of M2 cases are attributable to smoking. There were no or weak associations between smoking and increased AML risk for other FAB subtypes. The finding by this study of an association between smoking and FAB subtype M2 confirms a previously published report and suggests that earlier findings of no or weak smoking-AML associations may have been due to lack of subtype-specific analysis. PMID- 11882529 TI - Residential radon and lung cancer risk in a high-exposure area of Gansu Province, China. AB - In the general population, evaluation of lung cancer risk from radon in houses is hampered by low levels of exposure and by dosimetric uncertainties due to residential mobility. To address these limitations, the authors conducted a case control study in a predominantly rural area of China with low mobility and high radon levels. Included were all lung cancer cases diagnosed between January 1994 and April 1998, aged 30-75 years, and residing in two prefectures. Randomly selected, population-based controls were matched on age, sex, and prefecture. Radon detectors were placed in all houses occupied for 2 or more years during the 5-30 years prior to enrollment. Measurements covered 77% of the possible exposure time. Mean radon concentrations were 230.4 Bq/m(3) for cases (n = 768) and 222.2 Bq/m(3) for controls (n = 1,659). Lung cancer risk increased with increasing radon level (p < 0.001). When a linear model was used, the excess odds ratios at 100 Bq/m(3) were 0.19 (95% confidence interval: 0.05, 0.47) for all subjects and 0.31 (95% confidence interval: 0.10, 0.81) for subjects for whom coverage of the exposure interval was 100%. Adjusting for exposure uncertainties increased estimates by 50%. Results support increased lung cancer risks with indoor radon exposures that may equal or exceed extrapolations based on miner data. PMID- 11882530 TI - Methodological problems in the molecular epidemiology of tuberculosis. AB - In systematic studies of the molecular epidemiology of tuberculosis, DNA fingerprinting is used to estimate the fraction of incident cases attributable to recent transmission of Mycobacterium tuberculosis rather than reactivation disease and to identify risk factors for recent transmission. This approach is based on the premise that tuberculosis cases that share a DNA fingerprint are epidemiologically related while cases in which fingerprints are unique are due to remote infection that has reactivated. In this paper, the authors review the objectives and design of molecular epidemiologic studies of tuberculosis, describe current analytical approaches, and consider the impact of these different approaches on study results. Using data from a previously published investigation of the epidemiology of tuberculosis conducted from 1990 to 1993 among tuberculosis patients in New York City, New York, the authors show how selecting different measures of disease frequency, comparison groups, and sampling strategies may impact the results and interpretability of the study. They demonstrate ways to conduct sensitivity analyses of estimated results and suggest strategies that may improve the usefulness of this approach to studying tuberculosis. PMID- 11882531 TI - Comparison of telephone and postal survey modes on respiratory symptoms and risk factors. AB - Little is known about the comparability of postal and telephone survey modes in epidemiology. A cross-sectional, population-based study (n = 25,000) of lung disease was performed in 1998-1999 in two regions of Norway. Initial surveying was done by postal questionnaire. A 1% random sample (n = 171) of previous postal responders were resurveyed by telephone or cellular contact. The response rate was 67% on the telephone/cellular interview. Fewer incomplete answers were given by telephone than by mail. A lower prevalence was found by telephone for morning cough and exposure to passive smoking at work or home. Reproducibility was high for asthma, hay fever, wheezing, and attacks of breathlessness. Moderate reproducibility was seen for symptoms of chronic bronchitis. Reproducibility was low for indoor and work environment, although it was high for early life factors. Concordance coefficients were high for all continuous measures such as height, body weight, and pack-years. The authors conclude that the comparability between the postal and the telephone survey modes was good. The telephone survey mode gave more complete information. Survey mode may have a moderate effect on study results, depending on the specific questions asked. PMID- 11882532 TI - Stability of electrocardiographic classification pre- and postglucose challenge. AB - Because of validity concerns, electrocardiograms (ECGs) in epidemiologic studies are usually taken in fasting subjects. It would be preferable logistically to record ECGs throughout the day. The authors investigated the stability of ECGs taken while fasting and approximately 1 hour after a 75-g glucose load on the same morning in 89 older men and women who were participants in the Rancho Bernardo (California) Chronic Disease Study between 1984 and 1995. A reader blinded to this comparison classified ECGs using the Minnesota code and Whitehall criteria. Of 75 initially normal tracings, 27% changed to possible ischemia postglucose. Of 12 tracings initially indicating possible ischemia, two reverted to normal (kappa = 0.40, 95% confidence interval: 0.21, 0.59). The two tracings initially scored as probable ischemia remained in that category postglucose. More ECGs worsened than improved, and the variability pre- and postglucose was at least as great as that between clinic visits conducted 8 years apart. PMID- 11882533 TI - Surprises in European science policy. PMID- 11882534 TI - Mining the bibliome: searching for a needle in a haystack? New computing tools are needed to effectively scan the growing amount of scientific literature for useful information. PMID- 11882535 TI - The Nobel Prize and beyond: an interview with Sir Paul Nurse. Interview by Susan R. Owens. PMID- 11882536 TI - Harnessing the immune system to battle Alzheimer's: some of the most promising approaches to fight Alzheimer's disease aim to develop vaccines. PMID- 11882537 TI - Will they throw the bath water out with the baby? The US Congress is still debating whether to outlaw cloning humans. PMID- 11882538 TI - A right for family planning: the benefits of contraception for women's health and social status. PMID- 11882539 TI - The actin filament architecture: tightly regulated by the cells, manipulated by pathogens. International Titisee Conference on the actin cytoskeleton: from signalling to bacterial pathogenesis. PMID- 11882540 TI - Old dogs and new tricks: meeting on mechanisms of eukaryotic transcription. PMID- 11882541 TI - Histone acetylation: a switch between repressive and permissive chromatin. Second in review series on chromatin dynamics. AB - The organization of eukaryotic chromatin has a major impact on all nuclear processes involving DNA substrates. Gene expression is affected by the positioning of individual nucleosomes relative to regulatory sequence elements, by the folding of the nucleosomal fiber into higher-order structures and by the compartmentalization of functional domains within the nucleus. Because site specific acetylation of nucleosomal histones influences all three aspects of chromatin organization, it is central to the switch between permissive and repressive chromatin structure. The targeting of enzymes that modulate the histone acetylation status of chromatin, in synergy with the effects mediated by other chromatin remodeling factors, is central to gene regulation. PMID- 11882542 TI - Requirement of functional telomeres for metaphase chromosome alignments and integrity of meiotic spindles. AB - Telomerase deficiency in the mouse eventually leads to loss of telomeric repeats from chromosome ends and to end-to-end chromosome fusions, which result in defects in highly proliferative tissues. We show that telomere dysfunction resulting from telomerase deficiency leads to disruption of functional meiotic spindles and misalignment of chromosomes during meiotic division of oocytes in late-generation (G4) mice. However, oocytes from first-generation (G1) mice lacking telomerase showed no appreciable telomere dysfunction and exhibited chromosome alignment at the metaphase plates of meiotic spindles, in a manner similar to that of wild-type mouse oocytes. These findings suggest that telomerase does not directly influence chromosome alignment and spindle integrity. Rather, functional telomeres may be involved in mediating metaphase chromosome alignment and maintaining functional spindles during meiotic division. PMID- 11882543 TI - Modulation of ISWI function by site-specific histone acetylation. AB - Mutations in Drosophila ISWI, a member of the SWI2/SNF2 family of chromatin remodeling ATPases, alter the global architecture of the male X chromosome. The transcription of genes on this chromosome is increased 2-fold relative to females due to dosage compensation, a process involving the acetylation of histone H4 at lysine 16 (H4K16). Here we show that blocking H4K16 acetylation suppresses the X chromosome defects resulting from loss of ISWI function in males. In contrast, the forced acetylation of H4K16 in ISWI mutant females causes X chromosome defects indistinguishable from those seen in ISWI mutant males. Increased expression of MOF, the histone acetyltransferase that acetylates H4K16, strongly enhances phenotypes resulting from the partial loss of ISWI function. Peptide competition assays revealed that H4K16 acetylation reduces the ability of ISWI to interact productively with its substrate. These findings suggest that H4K16 acetylation directly counteracts chromatin compaction mediated by the ISWI ATPase. PMID- 11882544 TI - Posthaemorrhagic ventricular dilatation. AB - Posthaemorrhagic ventricular dilatation is the most serious direct complication of intraventricular haemorrhage after preterm birth. It results initially from multiple small blood clots throughout the cerebrospinal fluid channels impeding circulation and reabsorption. Management is difficult and new treatment approaches are needed. PMID- 11882546 TI - The clinical conundrum of neonatal seizures. AB - There is increasing evidence that neonatal seizures have an adverse effect on neurodevelopmental progression and may predispose to cognitive, behavioural, or epileptic complications later in life. However, given the uncertainty about the efficacy and toxicity of the commonly used anticonvulsants, when and how aggressively to treat such seizures is a difficult decision. PMID- 11882547 TI - Hypertension in the newborn baby. AB - Hypertension is rare in the newborn infant. It has a cumbersome definition and diagnosis, and screening is not justified using present definitions and technology. Thresholds for starting antihypertensive treatment in the first month of life are not clear, and the treatment is difficult, with idiosyncratic responses to drugs in neonates with varying renal and hepatic function. PMID- 11882548 TI - Skeletal development in premature infants: a review of bone physiology beyond nutritional aspects. AB - Bone development is usually seen as a process of bone mineral accretion or increase in bone mass, and treatment of bone disorders usually consists of attempts to maximise bone mass accumulation by nutritional means only. However, from a functional perspective, bones should not be as heavy as possible, but rather as stable as necessary. It is therefore important to create conditions that stimulate bones to become more stable. PMID- 11882549 TI - Perinatal risk factors for major intraventricular haemorrhage in the Australian and New Zealand Neonatal Network, 1995-97. AB - BACKGROUND: In 1995, large differences were identified in rates of grade 3-4 intraventricular/periventricular haemorrhage (major IVH) among neonatal intensive care units (NICUs) in the Australian and New Zealand Neonatal Network. AIMS: To develop a predictive model for major IVH in order to allow risk adjustment for the variation in rates of major IVH among NICUs. METHODS: Rates of IVH were determined in 5712 infants of 24-30 weeks gestation born from 1995 to 1997. Significant antenatal and perinatal variables for major IVH in 1995 and 1996 were identified by univariate and multivariate analysis. A predictive model was developed and then validated on 1997 data. RESULTS: Rates of all grades of IVH fell from 1995 to 1997 (30.4 to 24.3%) but wide interunit variation remained. Seven antenatal and perinatal characteristics had significant association with major IVH: fetal distress, intrauterine growth restriction (protective), antenatal corticosteroids (protective), gestational age, 1 minute Apgar <4, male gender, and transfer after birth. A predictive model based on the last five of these variables was developed using data from 1995 and 1996 which gave an area under the receiver operator characteristic (ROC) curve of 0.76. This model was then validated on the 1997 dataset where an identical ROC curve resulted. CONCLUSIONS: Antenatal and perinatal factors are important in the pathogenesis of major IVH. The predictive model developed from these factors can be used to adjust for confounders in interunit outcome comparison. PMID- 11882550 TI - Newborn screening for Duchenne muscular dystrophy: a psychosocial study. AB - OBJECTIVE: To evaluate the psychosocial implications of newborn screening for Duchenne muscular dystrophy. DESIGN: Prospective psychosocial assessment. SETTING: Primary care. RESPONDENTS: STUDY: (a) families of an affected boy identified by screening (n = 20); (b) families of a boy with a transient screening abnormality (n = 18). CONTROL: (a) families of a boy with a later clinical diagnosis (n = 16); (b) random sample of mothers of boys aged 6-9 months (n = 43). INTERVENTIONS: Questionnaires and semistructured interviews. MAIN OUTCOME MEASURES: Attitudes to newborn screening and impact of screening on mother-baby relationship, anxiety/wellbeing, and reproductive patterning within families of an affected boy. RESULTS: Most families of an affected boy were in favour of newborn screening on the grounds of reproductive choice and time to prepare emotionally and practically. There was no evidence of any long term disruption to the mother-baby relationship. Anxiety levels for the screened group were slightly above threshold but returned to normal during the period of the study. There was no evidence, from anxiety or wellbeing scores, that the transient group had suffered any disadvantage. Although the profile of the screened and later clinically diagnosed cohorts was similar after diagnosis, when boys from the screened cohort were 4 years old and more socially aware, their profile was more positive. There was evidence that reproductive patterning had been modified, and four fetuses carrying a mutation causing Duchenne muscular dystrophy were terminated. CONCLUSION: A case can be made for newborn screening provided that the test is optional, a rigorous protocol for service delivery is used, and an infrastructure providing continuing support is in place. PMID- 11882551 TI - Antenatal steroids are associated with a reduction in the incidence of cerebral white matter lesions in very low birthweight infants. AB - AIMS: To investigate whether antenatal steroids reduce the incidence of cerebral white matter lesions in very low birthweight infants. METHODS: A total of 224 newborn infants of < 31 weeks gestational age and weighing < 1500 g was studied between January 1998 and June 2000. Obstetric and neonatal information was obtained from the case notes. The study population was subdivided into two groups according to antenatal steroid exposure. A complete course of treatment consisted of two doses of 12 mg each of betamethasone given at an interval of 12-24 hours. Infants in group 1 were born to mothers who had not received betamethasone, or were delivered within 24 hours of receiving the first dose of steroid. Infants in group 2 were born to mothers who had received one or more complete courses of betamethasone and were delivered > 24 hours after receiving the first dose of steroid. RESULTS: The two groups contained statistically similar proportions of boys and girls, and the infants had similar birth weights and survival rates. Those in group 2, compared with those in group 1, had a lower gestational age (p = 0.02) and a lower incidence of white matter lesions on cranial ultrasound scans (p = 0.03). Stepwise logistic regression analysis showed that gestational age (p = 0.0002) and a complete course of antenatal steroids (p = 0.02) had independent effects on cerebral white matter lesions. CONCLUSIONS: These observations suggest that a complete course of antenatal steroids may have a protective effect against cerebral white matter lesions in very low birthweight infants. PMID- 11882552 TI - Follow up of a randomised trial of two different courses of dexamethasone for preterm babies at risk of chronic lung disease. AB - OBJECTIVES: To report 18 month outcome of a randomised trial of two courses of dexamethasone to prevent chronic lung disease of prematurity. STUDY DESIGN: Babies of birth weight 1250 g or less ventilated at 7 days of age were randomised to a 42 day reducing course (long) or a 3 day pulsed (pulse) course of dexamethasone. Growth, cardiovascular status, and respiratory and neurodevelopmental outcomes were assessed at 18 months. RESULTS: Seventy six babies were enrolled. Nine died and three were lost to follow up. Babies receiving the long course were weaned off oxygen more quickly than those receiving the pulse course (47% v 69% on oxygen at 28 days; p = 0.01), but there were no differences in 18 month outcomes. However, children averaged -1 SD for growth parameters, half had moderate or severe disability, and 35% and 19% respectively required oxygen at 36 weeks and discharge. CONCLUSIONS: The dexamethasone course used did not influence long term outcome. However, entry criteria for this study selected a group of babies at high risk of poor long term outcome. PMID- 11882553 TI - Outcome at school age following antenatal detection of absent or reversed end diastolic flow velocity in the umbilical artery. AB - AIM: To determine whether fetal compromise, manifested by abnormalities of Doppler recordings of umbilical artery velocity waveforms in utero, is associated with neurodevelopmental or educational abnormalities at school age. METHODS: A cohort of neonates born following high risk pregnancies had been previously identified for a study of the perinatal sequelae of absent (AEDFV) and reversed (REDFV) end diastolic flow velocities. Seventy six children were assessed at 5-12 years of age by a developmental paediatrician who was blinded to perinatal course and Doppler assessments. Forty children born following pregnancies with forward end diastolic flow velocities (FEDFV), were compared with 27 with AEDFV and nine with REDFV. Tests of cognitive, neurological, and sensory function were performed, and reports of behavioural and educational progress were obtained from parents and teachers. RESULTS: There were no significant differences between FEDFV and AEDFV groups, but on tests of mental ability and neuromotor function the REDFV group had worse scores than either FEDFV or AEDFV. Comparing REDFV and FEDFV groups, the British Ability Scales general conceptual ability mean scores were 87.7 versus 101, and the Quick Neurological Screening Test mean scores were 32.8 versus 21.5. CONCLUSIONS: Absence of EDFV is well recognised as a marker of fetal compromise which is associated with acute perinatal sequelae. This study suggests it is not associated with adverse neurodevelopmental outcome. However, we found reversal of EDFV on antenatal assessment to be associated with a wide range of problems at school age, suggesting that REDFV represents intrauterine decompensation which may have adverse effects on the developing brain. PMID- 11882554 TI - Do Not Resuscitate orders and ethical decisions in a neonatal intensive care unit in a Muslim community. AB - AIMS: To evaluate the need for Do Not Resuscitate (DNR) orders in a tertiary referral centre for neonatal intensive care, the criteria used in making these decisions, and the applicability of the Muslim ethical stance among parents in an Islamic community. METHODS: A prospective evaluation of all DNR decisions in the neonatal intensive care unit at the Royal Hospital in Oman, over a one year period between November 1999 and October 2000. This included decision criteria, and parental responses and expectations. RESULTS: Of 659 admissions to the neonatal intensive care unit during this period, DNR orders were written in 39 (6%) instances. Most related to congenital malformations (24/39, 62%). In those in whom ventilation was commenced (19/39, 49%) withdrawal was not culturally acceptable and expressly permitted in only 11%. For those in whom ventilation was not commenced (20/39, 51%), 70% agreed not to put their child on the ventilator if they did require it. Presence of extended family support (grandparents) and clergy was extremely useful. CONCLUSIONS: Asking parents alone to be explicitly involved or take full responsibility for decisions involving life and death is not culturally or socially acceptable in this community. Presence of extended family, and indirectly sounding out and taking into account their wishes, is more appropriate after assessing the resources and support services available. PMID- 11882556 TI - Measurement of the subarachnoid space by ultrasound in preterm infants. AB - BACKGROUND: Measurements of the subarachnoid space during routine cranial sonography may provide an indirect method of monitoring brain growth in preterm infants. METHODS: The width of the subarachnoid space was measured on coronal views during head sonography. Initial scans (within five days of birth) were compared with follow up scans. RESULTS: A total of 361 scans were performed on 201 preterm infants. The mean width of the subarachnoid space was < 3.5 mm for 95% of initial scans. It was slightly larger in neonates born closer to term, the equivalent of an increase of 0.02 mm/gestational week (95% confidence interval 0 to 0.10 mm) for initial scans. When the scans of all infants, born at 24-36 gestational weeks who were 36 weeks corrected gestational age were compared, the mean (SD) subarachnoid space was 60% larger for follow up scans than for initial scans: 3.2 (1.38) v 1.95 (1.35) mm (p = 0.002) or the equivalent of a mean increase of 0.20 mm/week (95% confidence interval 0.15 to 0.30 mm) for follow up scans. At 36 weeks corrected gestational age, mean head circumference was not different between those having initial or follow up scans (33.0 (2.0) v 32.2 (1.9) cm; p = 0.31). CONCLUSION: The mean subarachnoid space is normally < 3.5 mm in preterm infants. The difference between initial and follow up scans suggests reduced brain growth in extrauterine preterm babies. PMID- 11882555 TI - Effect of salt supplementation of newborn premature infants on neurodevelopmental outcome at 10-13 years of age. AB - BACKGROUND: The nutritional requirements of prematurely born infants are different from those of babies born at term. Inadequate or inappropriate dietary intake in the neonatal period may have long term adverse consequences on neurodevelopmental function. The late effect of neonatal sodium deficiency or repletion in the premature human infant on neurological development and function has not been examined, despite evidence in animals of a serious adverse effect of salt deprivation on growth of the central nervous system. METHODS: Thirty seven of 46 children who had been born prematurely (gestational age of 33 weeks or less) and allocated to diets containing 1-1.5 mmol sodium/day (unsupplemented) or 4-5 mmol sodium/day (supplemented) from the 4th to the 14th postnatal day were recalled at the age of 10-13 years. Detailed studies of neurodevelopmental performance were made, including motor function and assessment of intelligence (IQ), memory and learning, language and executive skills, and behaviour. Sixteen of the children were found to have been in the supplemented group and 21 in the unsupplemented group. RESULTS: Children who had been in the supplemented group performed better in all modalities tested than those from the unsupplemented group. The differences were statistically significant (analysis of variance) for motor function, performance IQ, the general memory index, and behaviour as assessed by the children's parents. The supplemented children outperformed the unsupplemented controls by 10% in all three components of the memory and learning tests (difference not significant but p < 0.1 for each) and in language function (p < 0.05 for object naming) and educational attainment (p < 0.05 for arithmetic age). CONCLUSION: Infants born at or before 33 weeks gestation require a higher sodium intake in the first two weeks of postnatal life than those born at or near term, and failure to provide such an intake (4-5 mmol/day) may predispose to poor neurodevelopmental outcome in the second decade of life. PMID- 11882557 TI - Deposition of whole blood platelets on extracellular matrix under flow conditions in preterm infants. AB - BACKGROUND: A previous study showed greater adhesion by platelets of healthy full term infants to subendothelial extracellular matrix (ECM) under flow conditions compared with healthy adult platelets. AIM: To investigate the adhesion and aggregation of platelets from preterm infants on ECM under defined shear conditions. METHODS: In vitro platelet function was investigated in 106 preterm infants, 74 full term infants, and 26 healthy adults. Blood samples were obtained from all infants within 24 hours of birth, and weekly until discharge from preterm infants only. Citrated whole blood was placed in ECM precoated tissue culture plates and subjected to shear stress (1300 s-1) for two minutes using a rotating Teflon cone. Platelet adhesion (surface coverage) and aggregation (average size) to ECM were assayed using an image analyser. Assays for von Willebrand factor (vWF) antigen, ristocetin cofactor, and vWF collagen-binding activity were performed on samples from an additional 70 preterm infants, 23 healthy full term infants, and 24 healthy adults. Preterm infants with hyaline membrane disease (HMD) were analysed separately in both cohorts. RESULTS: Platelets from preterm infants displayed significantly less platelet adhesion than those from full term infants but similar aggregation and levels of vWF antigen, ristocetin cofactor, and collagen binding activity. Mean surface coverage was 22.0 (8.4)% for preterm infants with HMD, 28.7 (8.0)% for healthy preterm infants, and 35.7 (7.9)% for full term infants. Surface coverage in the preterm infants correlated with gestational age during the first 24 hours only, and did not reach full term levels during 10 weeks of follow up. CONCLUSION: Platelet adhesion to ECM is significantly poorer in preterm than in full term infants, and poorer in preterm infants with HMD than in healthy preterm infants. Intrinsic platelet properties rather than the concentration or activity of vWF may be responsible for this difference. PMID- 11882558 TI - Effect of enteral administration of insulin on intestinal development and feeding tolerance in preterm infants: a pilot study. AB - OBJECTIVE: To determine in a pilot study whether enteral administration of insulin to preterm infants (26-29 weeks of gestational age) would enhance gastrointestinal development and reduce feed intolerance without adverse effects. DESIGN: Eight preterm infants were given 4 U/kg/day insulin enterally from 4 to 28 days of age. Lactase activity was measured at 28 days of age, while measures of feed intolerance were made throughout the hospital stay. The results were compared with those of a matched historical cohort of 80 preterm infants. SETTING: Tertiary care, university affiliated hospital. MAIN OUTCOME MEASURES: Lactase activity and feed intolerance. RESULTS: No adverse effects, such as hypoglycaemia, were observed after administration of insulin. The infants who received insulin had higher lactase activity and less feed intolerance than the controls (30% shorter time to full enteral feeds; fewer gastric residuals per infant). CONCLUSION: These preliminary data suggest that enteral insulin administration may be of benefit in reducing feed intolerance in preterm infants. A randomised, blinded trial is warranted. PMID- 11882559 TI - Congenital epiglottic cyst. PMID- 11882560 TI - Classical Kawasaki disease in a neonate. AB - A case is reported of classical Kawasaki disease in an infant younger than 2 weeks of age. Echocardiography detected a coronary artery aneurysm on the fifth day of the illness. Administration of intravenous gamma globulin resulted in rapid improvement. Kawasaki disease is rare in neonates, but it may follow a rapid and severe course. PMID- 11882561 TI - Stephane Tarnier (1828-1897), the architect of perinatology in France. AB - Stephane Tarnier studied medicine in Paris and became the doyen of obstetrics in France during the second half of the 19th century. He pioneered many advances and encouraged a perinatal approach to childbirth that was further developed by his disciples, Budin and Pinard. PMID- 11882562 TI - Prevention of postnatal cytomegalovirus infection in preterm infants. PMID- 11882563 TI - Correlation of umbilical cord weight with birth weight. PMID- 11882565 TI - Physiological genomics: implications in hypertension research. AB - In this article we delineate the directions in which the study of physiology will take as it becomes integrated with genomics. We also provide specific examples of the ways in which physiological genomics may be applied to study the complex genetics of hypertension and cardiovascular disease. PMID- 11882567 TI - Hypertension genetics, single nucleotide polymorphisms, and the common disease:common variant hypothesis. AB - The investigation of heritable susceptibility to disease is an effort to associate disease phenotype with underlying genotype. Such genotype:phenotype associations have been demonstrated for a large number of monogenetic disorders. The usual strategy has been to use linkage mapping in affected families to identify chromosomal loci from which candidate genes and genotypes can be tested for association with disease. This strategy has not been similarly successful for common heritable disease susceptibilities including hypertension that involve multiple genes and gene-environment interactions. Development of extensive collections of single nucleotide polymorphisms (SNPs) raises the possibility that these SNPs can be used as markers in genome-wide association mapping studies to identify hypertension susceptibility loci. In this approach, large numbers of markers are typed in cases and controls with the expectation that markers interrogating SNPs that are involved in inheritance of disease susceptibility will emerge through their association with this trait in the affected population. Essential hypertension is a common disorder. The term "common" has 2 implications: first, that the disease is prevalent; and, second, that it is widespread. Such frequency and distribution characteristics could arise if the susceptibility alleles for hypertension were prevalent in the founding population of contemporary human beings and became distributed with human global dispersal. This common disease:common variant concept is attractive because it suggests that the genetic heterogeneity underlying hypertension susceptibility could be relatively small. It also allows the possibility that nonrandom association of alleles (linkage disequilibrium, LD) can be used to reduce the number of SNP markers required to identify disease susceptibility alleles because a single marker can act as a surrogate for variation flanking it. The influence of a number of important factors on the detectability of hypertension susceptibility alleles by SNP mapping approaches is not yet fully defined. These factors include the locus and allelic diversity of hypertension, the weaker relationship (compared with Mendelian traits) between genotype and phenotype, the accuracy of high throughput genotyping techniques, the extensive role of nongenetic factors, and the extent and heterogeneous nature of LD across the genome. PMID- 11882568 TI - In search of cardiovascular candidate genes: interactions between phenotypes and genotypes. AB - Most cardiovascular traits of interest can be defined as "complex traits," with the first step in the identification of genetic factors affecting such traits being the detection of quantitative trait loci (QTLs). Animal models have proven particularly useful in this regard. However, only very few of the QTLs identified to date have led to the identification of candidate genes. We describe an example of our own work where the combination of anatomical and a biochemical intermediate phenotypes have led to the identification of the natriuretic peptide precursor A (Nppa) gene as a candidate gene for left ventricular hypertrophy (LVH). Combined with the power of comparative genetics, these strategies will continue to improve the chances of finding candidate genes for cardiovascular traits such as susceptibility to heart diseases, hypertension, and hypertension induced end-organ damage. PMID- 11882566 TI - Regulation of intrarenal angiotensin II in hypertension. AB - Intrarenal angiotensin II (Ang II) is regulated by several complex processes involving formation from both systemically delivered and intrarenally formed substrate, as well as receptor-mediated internalization. There is substantial compartmentalization of intrarenal Ang II, with levels in the renal interstitial fluid and in proximal tubule fluid being much greater than can be explained from the circulating levels. In Ang II--dependent hypertension, elevated intrarenal Ang II levels occur even when intrarenal renin expression and content are suppressed. Studies in Ang II--infused rats have demonstrated that augmentation of intrarenal Ang II is due, in part, to uptake of circulating Ang II via an Ang II type 1 (AT(1)) receptor mechanism and also to sustained endogenous production of Ang II. Some of the internalized Ang II accumulates in the light and heavy endosomes and is therefore potentially available for intracellular actions. The enhanced intrarenal Ang II also exerts a positive feedback action to augment intrarenal levels of angiotensinogen (AGT) mRNA and protein, which contribute further to the increased intrarenal Ang II in hypertensive states. In addition, renal AT(1) receptor protein and mRNA levels are maintained, allowing increased Ang II levels to elicit progressive effects. The increased intrarenal Ang II activity and AGT production are associated with increased urinary AGT excretion rates. The urinary AGT excretion rates show a clear relationship to kidney Ang II content, suggesting that urinary AGT may serve as an index of Ang II--dependent hypertension. Collectively, the data support a powerful role for intrarenal Ang II in the pathogenesis of hypertension. PMID- 11882569 TI - Discovery of a spontaneous genetic mouse model of preeclampsia. AB - Preeclampsia remains a leading cause of maternal and fetal morbidity and mortality but has an unknown etiology. Women with elevated baseline blood pressure have an increased risk of this disorder. We hypothesized that BPH/5 mice, an inbred mouse strain with mildly elevated blood pressure, would develop a pregnancy-induced hypertensive syndrome. Nonpregnant female BPH/5 and C57BL/6 mice underwent thoracic aortic implantation of telemeters. After 7 days of recovery and 5 days of baseline mean arterial blood pressure (MAP) recording, strain-matched timed matings were carried out. MAP was recorded continuously during pregnancy and for 1 week after birth. In separate mice in metabolic cages, urinary protein was tracked, followed by renal histological analysis. Before pregnancy, the BPH/5 strain had elevated baseline MAP compared with the C57BL/6 strain, but both strains had similar total urinary protein levels and renal histology. MAP remained stable in both groups during the first 2 weeks of pregnancy. However, at the start of the last trimester, MAP began to rise further in the BPH/5 mice; it rose to peak levels just before delivery and returned to prepregnancy levels by 2 days after delivery. This was accompanied by late gestational proteinuria and progressive glomerulosclerosis. No changes were observed in the C57BL/6 group except for a small decrease in MAP at mid gestation. The BPH/5 group delivered significantly smaller litters despite normal numbers of fetuses early in gestation, and longitudinal ultrasound studies documented fetal demise before the onset of hypertension and renal disease. This is the first report of an animal model that spontaneously develops a syndrome that bears close resemblance to preeclampsia, and it should have an impact on our understanding of the pathophysiology of this disorder. PMID- 11882570 TI - Renin-angiotensin system genetic polymorphisms and cerebral white matter lesions in essential hypertension. AB - It has been reported that both the DD genotype of the angiotensin converting enzyme (ACE) gene and the presence of cerebral white matter lesions (WML) may represent risk factors for the development of stroke. The present study investigates a possible association between 3 different genetic polymorphisms of the renin-angiotensin system and the presence of WML in 60 never-treated essential hypertensive patients (36 men, 24 women), aged 50 to 60 years, without clinical evidence of target organ damage. All patients underwent brain magnetic resonance imaging to establish the presence or absence of WML. The insertion/deletion (I/D) ACE gene, the M235T angiotensinogen (AGT) gene, and the A1166C angiotensin II type 1 receptor gene polymorphisms were determined by standard polymerase chain reaction. Twenty-five hypertensive patients (41.6%) were found to have WML on brain magnetic resonance imaging. No significant association between the M235T angiotensinogen or A1166C angiotensin II type 1 receptor genotypes and the presence of WML was found. However, the frequency of the DD genotype in patients with WML (64%) was significantly higher than that observed in patients without WML (28.6%; P=0.022). The DD genotype odds ratio for the presence of WML was 4.44 (95% confidence interval: 1.48 to 13.3). Likewise, the proportion of the D allele in patients with WML (74%) was significantly higher (P=0.014) than that observed in patients without WML (51.4%). We conclude that the presence of the DD genotype and/or the D allele of the ACE gene may be a predisposing factor for developing WML in essential hypertensive patients. However, because of the small sample size, these results require confirmation in a larger study. PMID- 11882571 TI - Heart rate and blood pressure quantitative trait loci for the airpuff startle reaction. AB - The airpuff startle reaction is a probe of sensori-autonomic processing and is useful for studies of genetic control of stress-induced cardiovascular activity. Using a Wistar-Kyoto-Spontaneously Hypertensive Rat F2 cross, we reported an airpuff-elicited strain-dependent and trial-dependent bradycardia, the absence of which cosegregated with hypertension. Here, we use the mapping power of the HXB BXH recombinant inbred rat strains (n=23) to locate quantitative trait loci (QTL) for this and associated cardiovascular phenotypes. Rats (12 weeks old), with indwelling femoral arterial catheters, were subjected to repeated airpuff startle stimuli (100 ms, 12.5 psi, 28 trials). Basal mean arterial pressure (MAP), delta MAP, and delta heart rate response to airpuff stimuli were analyzed as the average over 28 trials. There was a significant strain effect on the cardiovascular phenotypes measured. One QTL for the bradycardia elicited by the first airpuff stimulus was identified on chromosome 2 (D2rat 62/63; logarithm of odds [LOD] 2.9) mapping near a reported blood pressure locus. Further QTL were identified for basal MAP (RN08), stimulus-elicited tachycardia on trials 2 to 5 (RNO1 and RNO10), and delta MAP (RNO6). Our results indicate that chromosomes 1, 2, and 10 are involved in heart rate responses to airpuff startle stimulus, and chromosomes 6 and 8 are involved in pressor responses. This study is the first to identify stress-related heart rate loci and provides additional support for our prior cosegregation results. Furthermore, we have established the utility of this experimental paradigm to identify loci responsible for cardiovascular regulation during stress in genetic hypertensive models. PMID- 11882572 TI - The Y chromosome effect on blood pressure in two European populations. AB - Higher blood pressure (BP) in males compared with females is well documented and is thought to be influenced in part by the Y chromosome. To examine whether there is an association between BP and a polymorphic HindIII biallelic marker in the nonrecombining region of the Y chromosome, we genotyped 155 males from a Polish study group and 762 males from a Scottish study group. We also tested for possible interaction between the Y chromosome and a mutation in the steroidogenic factor binding site of the aldosterone synthase gene by genotyping the same group from Scotland. There was no significant difference in age or body mass index between 2 Y chromosome genotypes in both study groups. Men with the HindIII(+) genotype had significantly higher systolic and diastolic pressures than those with the HindIII(-) genotype in both the Polish and Scottish studies. This difference between the genotypes was 5.27 mm Hg (P=0.0014) and 3.14 mm Hg (P=0.0005) for adjusted systolic BP and 2.6 mm Hg (P=0.0045) and 1.44 mm Hg (P=0.0084) for adjusted diastolic BP in the Polish and the Scottish studied, respectively. On binary logistic regression analysis, males with the HindIII(+)/TT SF1 genotype combination had an odds ratio for elevated BP of 3.92 (CI 1.21 to 12.68, P=0.023). Our results indicate that the Y chromosome harbors a locus or loci that contribute to BP variation in hypertensive and normotensive men. The polymorphism in the aldosterone synthase gene may interact with the Y chromosome to increase the odds of an individual's developing higher BP. PMID- 11882573 TI - alpha-Adducin 460Trp allele is associated with erythrocyte Na transport rate in North Sardinian primary hypertensives. AB - Erythrocyte membrane alterations mirror those of vascular smooth muscle and renal tubular cell membrane. The interaction between adducin and Na-K pump is the most likely biochemical mechanism responsible for the increased tubular Na reabsorption and hypertension in Milan hypertensive strain (MHS) rats. To substantiate this hypothesis in humans, we tested to see if alpha-adducin Gly460Trp genotype is associated with erythrocyte sodium transport rate in a new cohort of n=268 never-treated North Sardinian primary hypertensives. Plasma renin activity and blood pressure response to hydrochlorothiazide were also measured to evaluate the relationship between sodium transport rate and two intermediate phenotypes with a higher degree of genetic complexity. Na-K pump, Na-K-Cl cotransport, and Li-Na countertransport at V(max) were faster (P<0.0001), whereas intracellular Na concentration was lower (P<0.0001) in patients carrying one or two 460Trp alleles. Such behavior was mirrored by opposite changes of intracellular Na concentration. Plasma renin activity and blood pressure response to diuretic treatment, on the other hand, showed a weaker association with the sodium transport rate. In conclusion, our findings are consistent with the hypothesis that the Gly460Trp alpha-adducin polymorphism may affect renal Na handling through an alteration in ion transport across the cell membrane mirrored by erythrocytes. These results may also have clinical relevance because the Gly460Trp alpha-adducin polymorphism may explain, at least in part, the variability of blood pressure response to diuretics in primary hypertensive patients. PMID- 11882574 TI - Phospholipase D contributes to transmural pressure control of prorenin processing in juxtaglomerular cell. AB - This study was designed to delineate the involvement of phospholipase C (PLC) and phospholipase D (PLD) in transmural pressure control of renin synthesis and secretion. Primary cultures of rat juxtaglomerular (JG) cells were applied to a transmural pressure-loading apparatus for 12 hours, and the renin secretion rate (RSR), active renin content (ARC), and total (active + inactive) renin content (TRC) were determined. Under control conditions (n=5), transmural pressure decreased RSR (78.1 +/- 3.0 and 64.6 +/- 4.4% for 0 or 40 mm Hg, respectively; P<0.05) and ARC (42.8 +/- 3.3 and 26.0 +/- 3.9 ng of angiotensin I per hour per million cells for 0 or 40 mm Hg, respectively; P<0.05) but did not have a significant effect on TRC (99.5 +/- 6.7 and 89.2 +/- 4.6 ng of angiotensin I per hour per million cells for 0 or 40 mm Hg, respectively). Treatment with PLC inhibitors, 2-nitro-4-carboxyphenyl-N,N-diphenyl-carbamate (200 micromol/L) and U73122 (10 micromol/L) did not alter RSR but prevented the RSR decrease with transmural pressure, whereas neither 2-nitro-4-carboxyphenyl-N,N-diphenyl carbamate nor U73122 altered ARC, TRC, or the decrease in ARC with transmural pressure. Experiments were also performed using JG cells (n=5) treated with a PLD inhibitor, 4-(2-aminoethyl)-benzensulfonyl fluoride (AEBSF, 100 micromol/L). Treatment with AEBSF did not influence basal levels of RSR, ARC, and TRC or the RSR decrease with transmural pressure. However, AEBSF did inhibit the decrease in ARC with transmural pressure. These results indicate that transmural pressure inhibits renin secretion via PLC-dependent pathways and prevents conversion of inactive renin to active renin via PLD-dependent mechanisms in JG cells. PMID- 11882575 TI - Low-salt diet enhances vascular reactivity and Ca(2+) entry in pregnant rats with normal and reduced uterine perfusion pressure. AB - Salt moderation is often recommended to prevent excessive increases in blood pressure during pregnancy, particularly in women who are prone to pregnancy induced hypertension; however, the vascular effects of low dietary salt intake during pregnancy are unclear. We investigated whether a low-salt diet during pregnancy alters the mechanisms of vascular smooth muscle contraction. Active stress and (45)Ca(2+) influx were measured in endothelium-denuded aortic strips of virgin and normal pregnant Sprague-Dawley rats and a hypertensive pregnant rat model produced by reduction in uterine perfusion pressure (RUPP), fed either a normal-sodium (NS, 1% NaCl) or low-sodium diet (LS, 0.2% NaCl) for 7 days. The mean arterial pressure was as follows: virgin/NS 108 +/- 8, virgin/LS 117 +/- 7, pregnant/NS 102 +/- 3, pregnant/LS 117 +/- 4, RUPP/NS 119 +/- 3, and RUPP/LS 133 +/- 6 mm Hg. Phenylephrine (Phe) caused concentration-dependent increases in active stress and (45)Ca(2+) influx that were greater in RUPP rats than in normal pregnant or virgin rats and were enhanced in pregnant/LS and RUPP/LS compared with pregnant/NS and RUPP/NS, respectively. High KCl (16 to 96 mmol/L), which stimulates Ca(2+) entry from the extracellular space, also caused increases in active stress that were greater in RUPP than in normal pregnant, in pregnant/LS than in pregnant/NS, and in RUPP/LS than in RUPP/NS rats. The Phe-induced (45)Ca(2+) influx--active stress relation was greater in RUPP/NS than in pregnant/NS and was enhanced in pregnant/LS and RUPP/LS compared with pregnant/NS and RUPP/NS, respectively. In Ca(2+)-free (2 mmol/L ethylene glycol bis(beta aminoethylether)-N,N,N',N'-tetra-acetic acid) Krebs, stimulation of intracellular Ca(2+) release by Phe (10(-5) mol/L) or caffeine (25 mmol/L) caused a transient contraction that was not significantly different in all groups of rats. Thus, a low-salt diet in pregnant and RUPP rats is associated with increases in vascular reactivity that involves Ca(2+) entry from the extracellular space but not Ca(2+) release from the intracellular stores. The enhancement of the Phe-induced Ca(2+) influx--active stress relation in pregnant and RUPP rats on a low-salt diet suggests activation of other vascular contraction mechanisms in addition to Ca(2+) entry. Although it is difficult to extrapolate the experimental data in rats to clinical data in women, the increased vascular reactivity and Ca(2+) entry and the possible enhancement of additional vascular contraction mechanisms with a low-salt diet suggest that reduction of dietary salt intake should be carefully monitored during pregnancy and pregnancy-induced hypertension. PMID- 11882576 TI - Effect of ACE inhibitors and angiotensin II type 1 receptor antagonists on endothelial NO synthase knockout mice with heart failure. AB - The beneficial effects of ACE inhibitors (ACEi) or angiotensin II type 1 receptor antagonists (AT(1)-ant) are reportedly mediated by NO in heart failure (HF). We hypothesized that in the absence of endothelial NO synthase (eNOS), (1) left ventricular (LV) dysfunction and myocardial remodeling would be more severe after myocardial infarction (MI), and (2) the cardioprotective effect of ACEi and AT(1) ant would be diminished or absent in mice with HF after MI. eNOS knockout mice (eNOS-/-) and wild-type C57BL/6J (C57) mice (+/+) were subjected to MI by ligating the left coronary artery. One month after MI, each strain was treated with vehicle, ACEi (enalapril, 20 mg/kg per day), or AT(1)-ant (valsartan, 50 mg/kg per day) for 5 months. Echocardiography was performed, and systolic blood pressure was measured before MI and monthly thereafter. Interstitial collagen fraction and myocyte cross-sectional area were examined histologically. We found that (1) compared with C57 mice, eNOS-/- mice that underwent sham surgery had significantly increased systolic blood pressure (P<0.05) and increased LV mass both initially and at 1 to 3 months, although cardiac function and histological findings did not differ between strains; (2) the development of HF and myocardial remodeling were similar after MI in both strains; and (3) ACEi improved cardiac function and remodeling in C57 mice, as evidenced by increased LV ejection fraction (LVEF) and LV shortening fraction (LVSF) and decreased diastolic LV dimension, mass, myocyte cross-sectional area, and interstitial collagen fraction, but these benefits were absent or diminished in eNOS-/- mice (for C57 versus eNOS-/-: increase in LVEF after ACEi, 14.2 +/- 2% versus -4.9 +/- 2.5%, respectively [P<0.001]; increase in LVSF, 8.6 +/- 2.1% versus -7.2 +/- 2.8%, respectively [P<0.01]; and decrease in LV mass, -16.6 +/- 15 versus 73 +/- 23 mm(3), respectively [P<0.01]). AT(1)-ant had benefits similar to those of ACEi, which were also absent or diminished in eNOS-/- mice (for C57 versus eNOS-/-: increase in LVEF after AT(1)-ant, 13.5 +/- 1.8% versus -9.8 +/- 3%, respectively [P<0.001]; increase in LVSF, 6.1 +/- 1.6% versus -3.8 +/- 3.1%, respectively [P<0.01]). Our data suggest that the absence of NO does not alter the development of HF after MI; however, it significantly decreases the cardioprotective effects of ACEi or AT(1)-ant. PMID- 11882577 TI - Trophic effects of the cyclooxygenase-2 product prostaglandin E(2) in cardiac myocytes. AB - Interleukin-1beta (IL-1beta), a proinflammatory cytokine, induces cyclooxygenase 2 (COX-2) in cultured neonatal ventricular myocytes (NVMs), resulting in the preferential production of prostaglandin E(2) (PGE(2)). To explain the preferential PGE(2) release by myocytes, we studied whether its specific synthase, PGE(2) synthase (PGES), is also induced by IL-1beta. Because COX-2 has been extensively associated with cell growth, we questioned whether PGE(2) plays a role in cardiac cell growth. IL-1beta--treated myocytes showed induction of PGES protein and mRNA by Western blot and reverse transcription--polymerase chain reaction, respectively. Immunofluorescence studies revealed perinuclear localization of COX-2 and PGES in IL-1beta--treated myocytes. Exogenous PGE(2) increased protein synthesis in NVMs, as indicated by a 1.6-fold increase in [(3)H]leucine incorporation, comparable to the known hypertrophic factor phenylephrine (1.6-fold). Because PGE(2) exerts different effects through 4 receptor subtypes (EP(1), EP(2), EP(3), and EP(4)), we investigated whether these receptors are functional in myocytes. Treatment of NVMs with the selective EP(1)/EP(3) agonist sulprostone significantly increased protein synthesis (1.7 fold), whereas the EP(1)/EP(2) antagonist AH6809 blocked this effect by 43%. In contrast, AH6809 had no effect on PGE(2)-induced protein synthesis. Regarding second messengers, sulprostone had no effect on cAMP, whereas PGE(2) increased it. We concluded that (1) PGE(2) production requires the induction of its specific synthase; (2) in myocytes, the inducible enzymes COX-2 and PGES are perinuclear; and (3) PGE(2) and sulprostone induce cardiac myocyte growth but seem to activate a different subset of EP receptors. PMID- 11882578 TI - Substance P in subtotal nephrectomy-salt hypertension. AB - We have previously demonstrated that calcitonin gene-related peptide (CGRP) plays a counterregulatory role in subtotal nephrectomy-salt (SN-salt) hypertension through an increase in vascular responsiveness to the dilator activity of this neuropeptide. Substance P (SP) is often co-localized with CGRP in perivascular sensory nerves. To determine the role and mechanism of action of SP in SN-salt hypertension, we induced hypertension in 4- to 6-week-old male Sprague-Dawley rats (n=8) by subtotal nephrectomy and 1% saline drinking water. Sham-operated rats were given either tap water (n=9) or 1% saline to drink (n=9). Eleven to 13 days after each protocol, all rats had intravenous (for drug administration) and arterial (for continuous monitoring of mean arterial pressure [MAP]) catheters surgically implanted and were studied in the conscious and unrestrained state. Baseline MAP was significantly elevated in the SN-salt rats (157 +/- 6 mm Hg) compared with tap water--fed controls (128 +/- 3 mm Hg) and 1% saline--fed controls (132 +/- 5 mm Hg). Vehicle administration did not alter the MAP in any group. In contrast, administration of spantide-II (0.2 micromol/L in saline), an SP receptor antagonist, significantly elevated the MAP in SN-salt rats (13.9 +/- 0.8 mm Hg) compared with the tap water (1.7 +/- 1.7 mm Hg) and 1% saline controls (2.0 +/- 1.9 mm Hg). SP mRNA and peptide levels in dorsal root ganglia were not significantly different between the 3 groups. Administration of exogenous SP (12 and 24 nmol center dot L(-1) center dot kg(-1) intravenously) resulted in a significantly greater decrease in MAP in the SN-salt rats compared with both control groups. Taken together, these data suggest that in SN-salt hypertension, SP plays a counterregulatory role in the absence of an increase in its neuronal expression, thereby suggesting that one possible mechanism of this compensatory vasodilator response is enhanced vascular reactivity to SP. PMID- 11882579 TI - Upregulation of arterial serotonin 1B and 2B receptors in deoxycorticosterone acetate-salt hypertension. AB - Previous studies have established a role for 5-hydroxytryptamine (5-HT)(2B) and 5 HT(1B) receptors in mediating enhanced contraction to serotonin (5-HT) in arteries from hypertensive deoxycorticosterone acetate (DOCA)-salt rats. To determine whether the observed increase in responsiveness was due to upregulation of 5-HT receptors, we used Western analysis to measure 5-HT(1B) and 5-HT(2B) receptor protein density. In endothelium-denuded aortas from hypertensive DOCA salt rats (mean systolic blood pressure 192 +/- 6 mm Hg), 5-HT(1B) and 5-HT(2B) receptor proteins were upregulated approximately 2-fold compared with the response in the aortas of sham-operated control rats (mean systolic blood pressure 119 +/- 2 mm Hg). Contraction to 5-HT(2B) receptor agonists was also enhanced in arteries from Wistar-Furth rats given DOCA and salt. This strain is relatively resistant to the hypertensive effects of DOCA and salt treatment. A common factor between the model of DOCA-salt hypertension and the DOCA-salt- treated Wistar-Furth rats is the presence of mineralocorticoids. Therefore, we tested the hypothesis that mineralocorticoids can upregulate 5-HT(1B) and 5 HT(2B) receptors. Aortas from normal Sprague-Dawley rats were incubated with aldosterone (100 nmol/L) for 8, 12, 24, and 48 hours. The expression of 5-HT(2B) and 5-HT(1B) receptor proteins was significantly increased (approximately 2- fold over vehicle treatment) by 8 hours. 5-HT(2B) and 5-HT(1B) receptors were upregulated by aldosterone in a concentration-dependent manner, and incubation with spironolactone (10 micromol/L) blocked this upregulation. These data support the conclusion that the increased expression of 5-HT(1B) and 5-HT(2B) receptors observed in arteries from DOCA-salt rats may be partially due to mineralocorticoids acting via the mineralocorticoid receptor to modulate gene expression. PMID- 11882581 TI - In vivo evidence for antioxidant potential of estrogen in microvessels of female spontaneously hypertensive rats. AB - In studies conducted in vitro, it has been demonstrated that estrogen has an antioxidant potential that may contribute to its protective effects on the cardiovascular system. However, the antioxidant effect of estrogen in vivo has not been demonstrated. To address this issue, in this study the effects of estrogen on oxidative stress were evaluated in microvessels studied in vivo. Oxidative stress was evaluated by using intravital microscopy in mesenteric arterioles from female spontaneously hypertensive rats (SHR) in physiological estrous (OE), ovariectomized (OVX), OVX treated with estradiol (E(2)), or estradiol + progesterone (E/P). The mesenteries were superfused with hydroethidine, a reduced and nonfluorescent precursor of ethidium bromide (EB). In the presence of reactive oxygen species, hydroethidine is transformed intracellularly in EB, which binds to DNA and can be detected by its red fluorescence. The percentage of EB-positive nuclei along the arteriolar wall in OVX (28.4 +/- 4.3) was significantly increased compared with OE (14.2 +/- 3.9; P<0.05). The OVX overproduction of oxyradicals was attenuated by E(2) (15.7 +/- 2.2) and E/P (14.8 +/- 0.8). Treatment with the superoxide dismutase mimetic MnTMPyP attenuated by 75% the oxidation of hydroethidine in both OE and OVX. Conversely, mannitol, that decomposes hydroxyl radical, and L-NAME, a nitric oxide synthase inhibitor, had no significant effects on hydroethidine oxidation. No differences on hydrogen peroxide plasma concentration were observed among the groups, suggesting that superoxide anion is the most likely oxyradical involved in the increased oxidative stress observed in OVX. The treatment of mesenteries with diphenyleneiodonium (DPI), an nicotinamide adenine dinucleotide phosphate (NADPH)-oxidase inhibitor, but not with oxypurinol, a xanthine-oxidase inhibitor, produced a significant reduction of oxyradical generation in OVX microvessels and a slight decrease in those from OE. Chronic treatment of female SHR with losartan caused similar decreases in oxyradicals in both OE and OVX, whereas diclofenac and verapamil had no effects. Together these data suggest that estrogen reduces superoxide anion bioavailability in vivo. The antioxidant effect of estrogen, which can contribute to a less pronounced endothelial dysfunction in female SHR, may be dependent on a direct modulatory action of estrogen on NADPH activity. PMID- 11882580 TI - Proprotein convertase PC5 regulation by PDGF-BB involves PI3-kinase/p70(s6) kinase activation in vascular smooth muscle cells. AB - We have recently demonstrated that furin, PC5, and PC7, members of the subtilisin/kexin-like mammalian proprotein convertases (PCs), are found in rodent aorta. These PCs have been identified to activate several growth factors, adhesion molecules and extracellular matrix compounds by endoproteolytic cleavage. In the present study, we investigated the regulation of PC5 in vascular smooth muscle cells (VSMCs) in vitro and in vivo. Stimulation of rat aortic VSMCs with platelet-derived growth factor (PDGF)-BB (20 ng/mL), angiotensin II (Ang II, 1 micromol/L), or 10% fetal calf serum (FCS) for 48 hours increased DNA synthesis, as assessed by proliferating cell nuclear antigen (PCNA) immunoblotting. PC5 was strongly upregulated by PDGF-BB and 10% FCS (both 8-fold, P<0.05), whereas Ang II had no effect on PC5 protein levels compared with controls. The PCs furin and PC7, which display a comparable subcellular localization and cleavage activity, were found in VSMCs, but their levels did not increase following PDGF-BB, Ang II, or FCS stimulation. Time-course analysis revealed a rapid increase in PC5 levels after 30 minutes of PDGF-stimulation of VSMCs. PDGF-stimulated PC5 induction was inhibited by the PI3-kinase inhibitor wortmannin, and by rapamycin, an inhibitor of mTOR/p70(s6)-kinase (both P<0.05). In contrast, the mitogen-activated protein kinase (MAPK)-pathway inhibitor PD98059 did not inhibit PDGF-stimulated PC5 induction. Immunocytochemistry and in situ hybridization revealed low PC5 protein and mRNA levels in intact rat aorta in vivo. After balloon injury, PC5 protein and mRNA levels were strongly increased in proliferating PCNA-positive VSMCs. The present data demonstrate that PC5 is upregulated during proliferation of VSMCs in vivo and in vitro. We show that PDGF-induced PC5 expression is PI3-kinase/p70(s6)-kinase dependent. Thus, growth factors regulate the proprotein convertase PC5, which may play an important role during VSMC growth. PMID- 11882582 TI - Methoxyestradiols mediate the antimitogenic effects of locally applied estradiol on cardiac fibroblast growth. AB - Estradiol inhibits cardiac fibroblast growth and may protect against cardiac remodeling associated with heart disease. However, the mechanisms by which estradiol attenuates cardiac fibroblast growth remain unclear. Because cardiac fibroblasts express cytochrome P450s (CYP450s) and catechol-O-methyltransferase (COMT) capable of converting estradiol to hydroxyestradiols and methoxyestradiols, respectively, and because hydroxyestradiols and methoxyestradiols (estradiol metabolites with little affinity for estrogen receptors) are potent inhibitors of cardiac fibroblast growth, we hypothesized that the antimitogenic effects of estradiol are mediated via hydroxyestradiols and/or methoxyestradiols. The inhibitory effects of estradiol (1 to 100 nmol/L) on serum-stimulated (3)H-thymidine incorporation (DNA synthesis), (3)H-proline incorporation (collagen synthesis), and cell number (proliferation) were enhanced (P<0.005) by CYP450 inducers 3-methylcholanthrene (10 micromol/L) and phenobarbital (10 micromol/L). Moreover, the inhibitory effects of estradiol were blocked by the CYP450 inhibitor 1-aminobenzotriazole (10 micromol/L) and the COMT inhibitors quercetin (10 micromol/L) and OR486 (10 micromol/L). In contrast to estradiol, the modulators of CYP450 and COMT were poor ligands for estrogen receptors (binding affinity less-than-or-equal 0.0001% versus estradiol). In cardiac fibroblasts, both quercetin and OR486 inhibited the metabolism of hydroxyestradiol to methoxyestradiol and blocked the inhibitory effects of hydroxyestradiol on cardiac fibroblast proliferation and DNA and collagen synthesis. The abrogating effects of quercetin and OR486 on the metabolism and antimitogenic effects of 2-hydroxyestradiol were mimicked by 20 micromol/L norepinephrine and isoproterenol, substrates for COMT. Our findings provide evidence that estradiol can inhibit cardiac fibroblast growth via an estrogen receptor--independent pathway that involves the local metabolism of estradiol to methoxyestradiols. PMID- 11882583 TI - Role of methoxyestradiols in the growth inhibitory effects of estradiol on human glomerular mesangial cells. AB - Metabolism of locally applied 17beta-estradiol (estradiol) to methoxyestradiols contributes to the growth inhibiting effects of estradiol on vascular smooth muscle cells via an estrogen receptor (ER)-independent mechanism. Because vascular smooth muscle cells are phenotypically similar to glomerular mesangial cells, it is feasible that estradiol inhibits glomerular mesangial cell growth via a similar mechanism, and this possibility was investigated. In human glomerular mesangail cells, estradiol concentration dependently (1 to 100 nmol/L) inhibited serum-induced proliferation (cell number) and DNA ((3)[H]-thymidine incorporation) and collagen ((3)[H]-proline incorporation) synthesis. The inhibitory effects of estradiol were mimicked by 2-hydroxyestradiol and 2 methoxyestradiol, metabolites of estradiol with little affinity for ERs. 2 Hydroxyestradiol and 2-methoxyestradiol were more potent growth inhibitors than estradiol. The inhibitory effects of estradiol were enhanced by CYP450 inducers 3 methylcholanthrene (10 micromol/L) and phenobarbital (10 micromol/L) and blocked by the CYP450 inhibitor 1-aminobenzotriazole (10 micromol/L). The growth inhibitory effects of estradiol were also blocked by quercetin (10 micromol/L) and OR 486 (10 micromol/L) inhibitors of catechol-O-methyltransferase (converts catecholestradiols to methoxyestradiols). ICI182780 (ER antagonist with ER binding affinity similar to estradiol) blocked the growth inhibitory effects of estradiol (1 to 100 nmol/L) only at concentrations (>50 micromol/L) that inhibited estradiol metabolism to catecholestradiols. The growth inhibitory effects of 2-hydroxyestradiol were abrogated by quercetin and OR486 (two structurally dissimilar catechol-O-methyltransferase inhibitors), but not by ICI182780. However, the growth inhibitory effects of 2-methoxyestradiol were unaltered by catechol-O-methyltransferase inhibitors and ICI182780. In conclusion, our findings provide the first evidence that methoxyestradiols mediate the growth inhibitory effects of locally applied estradiol on glomerular mesangial cell growth via an ER-independent mechanism. PMID- 11882584 TI - Gender differences in vascular smooth muscle reactivity to increases in extracellular sodium salt. AB - Hypertension is more common in men and postmenopausal women than in premenopausal women, and gender differences in sensitivity to high dietary Na(+) salt have been suggested; however, the vascular mechanisms involved are unclear. We investigated whether increases in the extracellular concentration of Na(+) ([Na(+)](e)) enhance the mechanisms of vascular smooth muscle contraction and whether the vascular effects of [Na(+)](e) exhibit gender differences. Isometric contraction and (45)Ca(2+) influx were measured in endothelium-denuded aortic strips that were isolated from intact male, intact female, castrated male, and ovariectomized (OVX) female Sprague-Dawley rats and incubated in Krebs' solution (2.5 mmol/L Ca(2+)) containing increasing [Na(+)](e) by the addition of 1, 3, 6, 10, 20, and 30 mmol/L NaCl. Increasing [Na(+)](e) for 30 minutes did not increase the resting tone or (45)Ca(2+) influx in any group of rats. Phenylephrine (Phe) caused concentration-dependent increases in contraction and (45)Ca(2+) influx. In vascular strips from intact males, increasing [Na(+)](e) by the addition of 1 to 6 mmol/L NaCl significantly increased the magnitude of Phe contraction and (45)Ca(2+) influx. Further increases in [Na(+)](e) by the addition of 10, 20, and 30 mmol/L NaCl increased Phe-induced (45)Ca(2+) influx but inhibited Phe contraction, possibly because of excessive increases in ionic strength. Preincubation with 2,4-dichlorobenzamil (10(-5) mol/L), inhibitor of the Na(+) Ca(2+) exchanger, or KB-R7943 (10(-5) mol/L), selective inhibitor of the reverse mode of the Na(+)-Ca(2+) exchanger, abolished the increases in Phe contraction and (45)Ca(2+) influx at increasing [Na(+)](e) obtained by the addition of 1 to 6 mmol/L NaCl. Preincubation in Krebs' solution containing control [Na(+)](e) plus 1 to 6 mmol/L LiCl or N-methyl-D-glucamine did not increase Phe contraction. In intact females, the Phe contraction and Ca(2+) influx were less than those in intact males and were not enhanced with increases in [Na(+)](e). The enhancement of Phe contraction and Ca(2+) influx with increases in [Na(+)](e) were not significantly different between castrated male rats and intact male rats but were greater in OVX female rats than intact female rats. In OVX female rats or castrated male rats treated with 17beta-estradiol (but not 17alpha-estradiol) subcutaneous implants, no significant changes in Phe contraction or Ca(2+) influx with increases in [Na(+)](e) were observed. In OVX female or castrated male rats simultaneously treated with 17beta-estradiol plus the estrogen receptor antagonist ICI 182,780, the Phe contraction and Ca(2+) influx were enhanced with increases in [Na(+)](e). Thus, in intact male rats, small physiological increases in [Na(+)](e) enhance smooth muscle contraction to Phe by a mechanism involving Ca(2+) entry, possibly via the reverse mode of the Na(+)-Ca(2+) exchanger. This mechanism appears to be reduced in the presence of endogenous or exogenous estrogen and thereby protects female rats against excessive increases in vascular reactivity during high dietary Na(+) intake. PMID- 11882585 TI - Pressure promotes angiotensin II--mediated migration of human coronary smooth muscle cells through increase in oxidative stress. AB - Angiotensin II--mediated oxidative stress may play a role in the pathogenesis of coronary atherosclerosis. We examined the effects of pressure on the angiotensin II--mediated increase in oxidative stress and migration of cultured human coronary smooth muscle cells (SMCs). Increased pressure (100 mm Hg) by helium gas for 48 hours increased angiotensin II--mediated oxidative stress as evaluated by flow cytometry and SMC migration (from 15.9 +/- 2.2 to 32.0 +/- 2.4 cells per 4 high-power fields, P<0.05; n=8). The pressure-induced increases in oxidative stress observed appear to involve phospholipase D (PLD) and protein kinase C (PKC), inasmuch as the indirect PLD inhibitor suramin, at 100 micromol/L, and the PKC inhibitor chelerythrine, at 1 micromol/L, completely blocked the increase in angiotensin II--mediated oxidative stress induced by pressure. Pressure-induced increase in angiotensin II--mediated oxidative stress was inhibited by diphenylene iodonium chloride, an NADPH oxidase inhibitor, by 79% (P<0.05, n=8). Losartan (1 micromol/L), its active metabolite E3174 (1 micromol/L), and the antioxidant N-acetylcysteine (100 mmol/L) but not PD123319 (1 micromol/L) also blocked pressure-induced increases in angiotensin II--mediated oxidative stress and SMC migration (P<0.05, n=8). These findings suggest a novel cellular mechanism whereby pressure regulates the angiotensin II--mediated migration of SMCs, possibly via angiotensin II type 1 receptors, and which involves PLD mediated, PKC-mediated, and NADPH oxidase--mediated increases in oxidative stress. PMID- 11882586 TI - Nitric oxide induces dilation of rat aorta via inhibition of rho-kinase signaling. AB - NO induces vasodilation through cGMP-dependent protein kinase--dependent and - independent mechanisms. A recent study demonstrated that recombinant cGMP dependent protein kinase can phosphorylate the small G protein, RhoA, thus inhibiting its activity. Additionally, sodium nitroprusside was found to reverse the phenylephrine-induced translocation of RhoA, which is further indicative of the inhibition of RhoA activity. RhoA is known to be involved in the Ca(2+) sensitization of vascular smooth muscle through the actions of one of its downstream effectors, Rho-kinase. This study examined whether NO endogenously induces the relaxation of intact rat aorta via the inhibition of the Rho-kinase- mediated Ca(2+)-sensitizing pathway. Endogenous Rho-kinase inhibitor activity was inhibited by the selective compound Y-27632. Treatment of endothelium-intact rat aorta with Y-27632 (1 micromol/L) resulted in an attenuation of maximal force generated in response to phenylephrine. In endothelium-denuded rings, however, 1 micromol/L Y-27632 was ineffective at inhibiting the phenylephrine-induced contraction. Additionally, 1 micromol/L Y-27632 was significantly less effective at inhibiting the phenylephrine-induced contraction of endothelium-intact rings in the presence of inhibitors of NO synthase or guanylate cyclase (N(omega)-nitro L-arginine and 1H-[1,2,4]oxadiazolo-[4,3-a]quinoxalin-1-one, respectively). Interestingly, sodium nitroprusside restored the ability of 1 micromol/L Y-27632 to attenuate phenylephrine-induced contraction. Rho-kinase inhibition was also found to increase the sensitivity of the endothelium-denuded aorta to sodium nitroprusside. These data demonstrate that NO inhibits Rho-kinase activity in the intact rat aorta, supporting the hypothesis that endogenous NO-mediated vasodilation occurs through the inhibition of Rho-kinase constrictor activity in the intact rat aorta. PMID- 11882587 TI - Implications of hyperhomocysteinemia in glomerular sclerosis in hypertension. AB - Hyperhomocysteinemia (hHcys) has been recognized as a new risk factor for cardiovascular diseases independent of plasma lipid levels or other factors. However, it remains unknown whether hHcys is implicated in the target organ damages associated with hypertension. The present study first examined the possible role of hHcys in the development of glomerulosclerosis in Dahl salt sensitive (DS) hypertensive rats. High-performance liquid chromatography showed that plasma total homocysteine (tHcys) concentration was 7.64 +/- 0.29 micromol/L in conscious DS rats on a low salt (0.4% NaCl) diet, which was higher than 5.23 +/- 0.25 micromol/L in Dahl salt-resistant normotensive rats. When these rats were exposed to a high salt (4% NaCl) diet, plasma tHcys markedly increased in DS rats (14.7 +/-1.31 micromol/L) but not in Dahl salt-resistant rats (5.34 +/- 0.54 micromol/L). An iron chelater, desferrioxamine (0.3 mg/kg IV per day), completely normalized high salt--induced elevations of plasma tHcys and significantly attenuated the sclerotic changes in the glomeruli in DS rats. To further determine whether hHcys has an independent effect in the development of glomerulosclerosis, Sprague-Dawley rats were fed drinking water containing methionine (1 g/kg per day) for 6 weeks to produce hHcys. In these rats, plasma tHcys increased to 12.5 +/- 1.9 micromol/L (versus 6.1 +/- 2.6 micromol/L in control rats), and the aorta exhibited typical sclerotic changes, but arterial pressure was not altered. Urinary protein excretion increased to 52 +/- 2 mg/24 hours (versus 17 +/- 2 mg/24 hours in control rats), and the glomerular mesangium was expanded with glomerular hypercellularity, capillary collapse, and fibrous deposition in the rats with hHcys. These results suggest that elevated plasma homocysteine may be an important pathogenic factor for glomerular damage in hypertension independent of arterial pressure. PMID- 11882588 TI - Lipocalin-type prostaglandin d synthase in essential hypertension. AB - Lipocalin-type prostaglandin D synthase (L-PGDS) reportedly well predicts cardiovascular injuries in humans. However, little is known about the implications of L-PGDS in hypertension. In the present study, we investigated the alterations of serum and urinary L-PGDS in hypertensive patients with or without renal dysfunction. A total of 111 patients with hypertension (EHT; 65 with normoalbuminuria, 23 with microalbuminuria, 12 with macroalbuminuria, 11 with renal failure) and 102 normotensive, nomoalbuminuric subjects (NT) were studied. L-PGDS was measured by enzyme-linked immunosorbent assay, and L-PGDS in the kidney was localized using immunohistochemical methods. Blood pressure was higher in EHT groups than in the NT group (P<0.0001). There were no differences in age, gender, BMI, TC, TG, and HbA1c levels among the groups. Serum creatinine and urinary albumin levels were higher in the group with renal failure. Serum levels of L-PGDS were increased in EHT with normoalbuminuria, as compared with NT (0.88 +/- 0.05 versus 0.65 +/- 0.02 microg/mL; P<0.001). Serum levels of L-PGDS increased with the renal function worsened and positively correlated with serum creatinine, particularly in patients with renal impairments (r=0.76, P<0.0001). Similarly, the urinary L-PGDS excretions in EHT with normoalbuminuria were higher than that in NT (2.31 +/- 0.29 versus 1.16 +/- 0.14 mg/gCr, P<0.001), whereas there were no differences in urinary albumin excretion between the 2 groups. Moreover, urinary L-PGDS excretion increased dramatically with an increase in albuminuria or proteinuria. L-PGDS was stained in the tubules and the interstitium of the kidney in nephrosclerosis. In conclusion, patients with hypertension exhibited a higher level of L-PGDS in serum and urine, and this became increasingly obvious along with advance in renal dysfunction. These data suggest that L-PGDS metabolism is related to blood pressure and kidney injuries associated with hypertension. PMID- 11882589 TI - Coexistence of hypercholesterolemia and hypertension impairs adventitial vascularization. AB - We have shown that adventitial vasa vasorum (AVV) formation is enhanced in hypertensive rat aorta to compensate hypoxia in the thickened media and that hypercholesterolemia impairs angiogenesis in rat ischemic hindlimb. Thus, we examined the effects of coexistence of hypercholesterolemia and hypertension on AVV formation. In Wistar rats, hypercholesterolemia was established by high cholesterol diet from Day -14 (HC rats), and hypertension was induced by a suprarenal aortic constriction at Day 0 (HT rats). At Day 28, we studied AVV density, adventitial area, and medial thickness in the ascending aorta of control (standard diet+sham operation), HC, HT, and HC+HT rats (n=5/group). In HC rats, although the adventitial area was modestly increased, the AVV density and medial thickness were unchanged versus controls. In addition to medial thickening, marked enlargement of the adventitial area accompanied by increased AVV density was observed in HT rats, compared with controls. HC+HT rats showed lower AVV density, despite larger adventitial area, than HT rats, whereas the medial thickness was similar in HT and HC+HT rats. Immunohistostaining revealed hypoxia inducible factor-1alpha expression in the media only in HC+HT rats but not in the other 3 groups, suggesting persistent medial hypoxia in HC+HT rats. In conclusion, it is suggested that coexistence of hypercholesterolemia and hypertension impairs AVV formation, resulting in insufficient compensation for hypoxia in the thickened media. Our findings provide an insight into the mechanism of the aggravation of arteriosclerosis when both hypercholesterolemia and hypertension are present. PMID- 11882590 TI - Dipeptidyl peptidase IV activity in patients with ACE-inhibitor-associated angioedema. AB - Bradykinin and substance P have been implicated as mediators in angiotensin converting enzyme inhibitor (ACEI)-associated angioedema. Studies investigating the metabolism of bradykinin in sera from patients with a history of ACEI associated angioedema and controls suggest that there is a defect in a non-ACE, non-kininase I pathway of bradykinin degradation, such as the aminopeptidase P (APP)/dipeptidyl peptidase IV (DPPIV) pathway. This study tested the hypothesis that serum APP or DPPIV activity is decreased in patients with ACEI-associated angioedema. APP and DPPIV activity were measured in sera collected from patients during ACEI-associated angioedema, from patients with a remote history of ACEI associated angioedema, and from normotensive and untreated hypertensive controls. The effects of acute and chronic ACEI and corticosteroid treatment on serum DPPIV activity were also assessed. DPPIV activity was similar in normotensive volunteers (37.8 +/- 6.3 nmol/mL per min), in untreated hypertensive subjects who had been exposed previously to ACEI without angioedema (36.2 +/- 4.3 nmol/mL per min), in hypertensive patients with a remote history of angioedema (35.1 +/-8.5 nmol/mL per min), and in chronically ACEI-treated hypertensive subjects (36.1 +/- 5.6 nmol/mL per min). DPPIV activity decreased with increasing age (R(2)=0.10, P=0.016). Subject group significantly affected DPPIV activity (F=6.208, P=0.016) such that DPPIV activity was significantly lower in patients with ACEI-associated angioedema (26.9 +/- 4.1 nmol/mL per min) than in normotensive controls, in previously ACEI-exposed untreated hypertensive volunteers, or in ACEI-treated hypertensive volunteers, even after controlling for age. There was no effect of acute ACE inhibition or corticosteroids on DPPIV activity. With respect to APP activity, there was no difference between groups. These results suggest that DPPIV activity is depressed in individuals with hypertension during acute ACEI associated angioedema. PMID- 11882591 TI - Vasopressin increases intracellular NO concentration via Ca(2+) signaling in inner medullary collecting duct. AB - The present study was designed to determine whether arginine vasopressin (AVP) stimulates NO production in the epithelial collecting duct cells of the inner medulla (IMCDs) and if this is mediated through Ca(2+) signaling. Thin tissue layers containing IMCDs were dissected from Sprague-Dawley rats. Intracellular Ca(2+) concentration ([Ca(2+)](i)) and NO production were measured in IMCDs by a fluorescence imaging system with the use of fura 2-AM and the cell-permeable form of the NO-sensitive dye 4,5-diaminofluorescein (DAF-2), respectively. AVP (100 nmol/L) produced a rapid peak increase in [Ca(2+)](i) of 320 +/- 70 nmol/L within a few seconds and a sustained increase of 120 +/- 62 nmol/L. The peak increase in [Ca(2+)](i) was followed by a significant increase of NO production (34 +/- 7 U). This was similar to that produced by 20 micromol/L of the NO donor DETA-NONOate (42 +/- 11 U). The NO scavenger carboxy-PTIO (100 micromol/L) or depletion of [Ca(2+)](i) by preincubation with 5 micromol/L of the Ca(2+)-ATPase inhibitor thapsigargin in Ca(2+)-free buffer abolished the NO response to AVP. We conclude that AVP mobilizes Ca(2+) to produce NO in IMCDs. PMID- 11882592 TI - Targeted proteomic profiling of renal Na(+) transporter and channel abundances in angiotensin II type 1a receptor knockout mice. AB - The renal tubule transporters responsible for Na(+) and water transport along the nephron have been identified and cloned, permitting comprehensive analysis of transporter protein abundance changes in complex physiological models by using a "targeted proteomics" approach. Here, we apply this approach to screen renal homogenates from mice in which the gene for the angiotensin II type 1a (AT(1a)) receptor has been deleted (versus wild-type mice) to determine which sodium transporters and channels are regulated by the AT(1a) receptor at the protein abundance level. In mice maintained on a low NaCl diet (<0.02% NaCl), (1) the abundances of 2 aldosterone-regulated transporters were markedly decreased in knockout versus wild-type mice, namely, the thiazide-sensitive cotransporter and the alpha-subunit of the amiloride-sensitive Na(+) channel (alpha-ENaC); (2) the abundances of beta-ENaC and gamma-ENaC were markedly increased; and (3) there were no significant changes in the abundances of the proximal tubule Na+-H(+) exchanger or the Na(+)-K(+)-2Cl(-) cotransporter of the thick ascending limb. When the experiment was repeated on higher NaCl diets (0.4% or 6% NaCl), the decrease in alpha-ENaC abundance persisted, whereas the other changes were abolished. Analysis of serum aldosterone concentration in AT(1a) knockout mice and wild-type mice on the low NaCl diet revealed the absence of a decrease with AT(1a) gene deletion (11.8 +/- 2.3 nmol/L for knockout mice and 5.7 +/- 0.8 nmol/L for wild-type mice [significantly increased]). These results reveal that the AT(1a) receptor plays an important role in regulation of Na(+) transporter and channel proteins in the "post-macula densa" region of the renal tubule via a mechanism that is not dependent on altered circulating aldosterone concentrations. PMID- 11882593 TI - Intracellular angiotensin II stimulates voltage-operated Ca(2+) channels in arterial myocytes. AB - Although the presence of intracellular angiotensin II (Ang II) and of Ang II binding sites has been reported, their roles in cell function have not been fully clarified. The purpose of the present study was to test the hypothesis that intracellular Ang II modifies voltage-operated Ca(2+) channels in vascular smooth muscle. Ca(2+) channel currents were recorded in guinea pig mesenteric arterial myocytes with the whole-cell patch-clamp method. Intracellular dialysis of Ang II increased the amplitudes of Ca(2+) channel current (133 +/- 9% of the control with 10 nmol/L Ang II, n=16). Concomitant dialysis of the Ang II type 1 receptor antagonist, CV-11974 (1 micromol/L, n=11), but not the bath application of this drug, suppressed this Ang II action. In contrast, the dialysis of the Ang II type 2 receptor antagonist, PD123319 (1 micromol/L, n=5), failed to affect the Ang II action. Dialysis of either a phospholipase C inhibitor (U-73122, 10 micromol/L, n=5) or protein kinase C inhibitors (calphostin C, 100 nmol/L, n=5; protein kinase C inhibitor peptide-[19-36], 1 micromol/L, n=5) suppressed the Ang II action. Dialysis of KT5720 (100 nmol/L, n=5), an inhibitor of cAMP-dependent protein kinase, did not affect the Ang II action. Intracellular dialysis of angiotensin I (10 nmol/L) enhanced Ca(2+) channel currents (13 3 +/- 8%, n=6), which were sensitive to intracellular enalaprilat (1 micromol/L, n=5) or CV-11974 (n=5). These results suggest that intracellular Ang II has a stimulating action on voltage-operated Ca(2+) channels in vascular smooth muscle, possibly through intracellular binding sites similar to the Ang II type 1 receptor, which are associated with phospholipase C and protein kinase C. PMID- 11882594 TI - Increased angiotensin II-mediated Src signaling via epidermal growth factor receptor transactivation is associated with decreased C-terminal Src kinase activity in vascular smooth muscle cells from spontaneously hypertensive rats. AB - We investigated whether upregulation of Src by Ang II leads to increased extracellular signal-regulated kinase 1/2 (ERK1/2) phosphorylation in vascular smooth muscle cells (VSMCs) from spontaneously hypertensive rats (SHR) and whether these processes are associated with altered activation of C-terminal Src kinase (Csk), a negative regulator of Src. Furthermore, the role of epidermal growth factor receptor (EGFR) transactivation by angiotensin II (Ang II) was determined. Ang II-mediated c-Src phosphorylation was significantly greater (approximately 4-fold, P<0.01) in SHR than in Wistar-Kyoto rats (WKY). Ang II increased Csk phosphorylation 2-to 3-fold in WKY but not in SHR. Treatment of the cells with AG1478, a selective EGFR tyrosine kinase inhibitor, decreased Ang II mediated c-Src phosphorylation, particularly in SHR. Phosphorylation of cortactin and Pyk2/focal adhesion kinase, Src-specific substrates, was increased by Ang II >3-fold, with significantly greater responses in SHR than in WKY (P<0.05). Ang II induced ERK1/2 activation was significantly augmented (P<0.05) and sustained in VSMCs from SHR. PP2, a selective Src inhibitor, attenuated these effects and normalized the responses in SHR. Irbesartan, a selective Ang II type 1 receptor blocker, but not PD123319, a selective Ang II type 2 receptor blocker, inhibited Ang II actions. Our results demonstrate that c-Src phosphorylation and Src dependent ERK1/2 signaling by Ang II are increased in VSMCs from SHR. These processes are associated with blunted Ang II-induced phosphorylation of Csk. EGFR transactivation contributes to Ang II-mediated Src-dependent ERK1/2 signaling. In conclusion, altered regulation of Ang II type 1 receptor-activated c-Src by Csk may be an important upstream modulator of abnormal ERK1/2 signaling in VSMCs from SHR. PMID- 11882595 TI - Selective resistance to central neural administration of leptin in agouti obese mice. AB - We recently demonstrated that in a rodent model of obesity (agouti yellow mice), there is a selective leptin resistance with preservation of the sympathetic actions despite loss of appetite and weight-reducing actions of systemic leptin. Here, we examined whether selective leptin resistance exists in agouti mice during central neural administration of leptin. In agouti obese mice and lean controls, we tested the effects of single intracerebroventricular (ICV) injection of leptin or vehicle on food intake and body weight in the conscious state and on renal sympathetic nerve activity during anesthesia. Agouti obese mice had higher (P<0.0001) mean arterial pressure (100 +/- 2 mm Hg) than lean controls (90 +/- 2 mm Hg). In lean controls (n=9 to 10), ICV leptin caused a dose-dependent decrease in body weight (P<0.001) and food intake (P<0.001). For example, ICV leptin (3 microg) decreased food intake and body weight, respectively, by 3.3 +/- 0.3 g (P<0.001) and 2.6 +/- 0.3 g (P<0.001) in lean mice. However, in agouti obese mice (n=9 to 10), ICV leptin did not significantly decrease food intake or body weight. ICV leptin caused in RSNA a significant and dose-dependent increase in renal sympathetic nerve activity that was of the same magnitude in the lean and agouti obese mice. The rise in renal sympathetic nerve activity induced by ICV leptin (3 microg) was 274 +/- 67% (P<0.001) in lean controls and 275 +/- 46% (P<0.001) in the agouti obese mice. In summary, this study indicates that selective leptin resistance in agouti obese mice occurs with central neural administration of leptin, suggesting that selective leptin resistance in this model is not due to a defect in leptin transport across the blood brain barrier. It seems to result instead from alterations in the central neural pathways mediating effects of leptin. PMID- 11882596 TI - Thyrotropin-releasing hormone decreases leptin and mediates the leptin-induced pressor effect. AB - Leptin, an adipocyte-released hormone, modifies food intake and energy expenditure regulating hypothalamic-pituitary-thyroid axis function. We previously reported that thyrotropin-releasing hormone (TRH) precursor gene overexpression induces hypertension in the normal rat and that spontaneously hypertensive rats have central TRH hyperactivity with increased TRH synthesis and release and an elevated TRH receptor number. In both models, intracerebroventricular antisense (AS) treatment against the TRH precursor produced a dose-dependent reduction of the increased diencephalic TRH content while normalizing high arterial blood pressure. In this article, we report that male Wistar rats that were made hypertensive by intracerebroventricular injection of a eucaryotic expression plasmid containing the pre-TRH cDNA showed decreased leptin plasma levels and that pre-TRH AS treatment reversed this phenomenon. In addition, male and female spontaneously hypertensive rats showed lower levels of circulating leptin than did sex-matched Wistar-Kyoto control rats. This difference also was abated by the pre-TRH AS treatment. Conversely, 20 microg ICV leptin induced a long-lasting pressor effect (18 +/- 5 mm Hg, n=6, P<0.01, >60 minutes) that was not observed in pre-TRH AS pretreated rats (2 +/- 3 mm Hg, n=6) but persisted in rats used as controls that were treated with inverted oligonucleotide (20 +/- 6 mm Hg, n=4, P<0.01). These data suggest that in rats with TRH-induced hypertension, leptin is decreased, inducing compensatory adiposity. We propose that because leptin produces central TRH synthesis and release, obesity may induce hypertension through TRH system activation and that the TRH-leptin interaction may thus contribute to the strong association between hypertension and obesity. PMID- 11882597 TI - Chronic cardiovascular and renal actions of leptin: role of adrenergic activity. AB - This study was designed to determine the role of changes in adrenergic activity in mediating the chronic cardiovascular, renal, and metabolic actions of leptin. Male Sprague-Dawley rats were implanted with catheters for mean arterial pressure (MAP) and heart rate (HR) measurements and IV infusions of either vehicle (n= 7) or alpha- and beta-adrenergic receptor antagonists, terazosin and propranolol (10 mg/kg/d; n= 8) throughout the study. After control measurements, murine leptin was infused IV (1.0 microg/kg/min) for 7 days along with vehicle or adrenergic antagonists, followed by a 7-day recovery period. Leptin infusion significantly reduced food intake in control rats from 22.6 +/- 0.8 to 10.6 +/- 0.4 g/d and, in adrenergic blockade rats, from 22.6 +/- 0.8 to 13.2 +/- 0.8 g/d. Fasting plasma insulin decreased from 48 +/- 10 to 5 +/- 2 microU/mL in control rats and from 51+/- 9 to 9 +/- 2 microU/mL in adrenergic blockade rats during leptin infusion. Leptin infusion did not significantly alter glomerular filtration rate in either group. MAP and HR increased by 6 +/- 1 mm Hg and 23 +/- 7 bpm after 7 days of leptin infusion in control rats. However, in adrenergic blockade rats, leptin infusion did not significantly alter MAP (-1 +/- 1 mm Hg) and decreased, rather than increased, HR (-23 +/- 8 bpm). These results indicate that leptin-induced increases in blood pressure and tachycardia are mediated by increased adrenergic activity and support the concept that leptin may be an important link between obesity, increased sympathetic activity, and hypertension. However, the chronic effects of leptin on insulin and glucose regulation do not appear to be altered by alpha- and beta-adrenergic receptor blockade. PMID- 11882598 TI - Src autophosphorylation is an early event in pressure-mediated signaling pathways in isolated resistance arteries. AB - Elevated blood pressure is associated with varying degrees of arterial growth and remodeling. The mechanisms by which mechanical stress is converted into cellular alteration have yet to be fully elucidated. Our laboratory has demonstrated that Src tyrosine kinases and the extracellular signal-regulated kinase subtype of the mitogen-activated protein kinase family mediate pressure-induced c-fos expression in rat mesenteric arteries. Others have reported involvement of integrin and growth factor receptor signaling pathways. Our goal was to determine the role of Src, focal adhesion kinase (FAK), and platelet-derived growth factor (PDGF) receptor signaling in the upstream initiation of these events. Pairs of rat mesenteric arteries were pressurized to 90 mm Hg (control), and then one was raised to 140 mm Hg for 1, 3, or 5 minutes. Western blotting revealed that Src pY(418) was elevated 2.4-fold over control values at 1 minute and 2.8-fold at 3 minutes and returned to control at 5 minutes. Significant FAK-Y(397) phosphorylation was observed only after 3 and 5 minutes of pressure stimulus and was blocked entirely by Src inhibition. Src-pY(215) activity (associated with PDGF receptor activation) does not seem to be involved at any of the time points tested. These data demonstrate that Src-Y(418) autophosphorylation is an early event in pressure mechanotransduction and leads to activation of FAK-Y(397). This finding suggests that Src may be the messenger that initiates and propagates the cellular growth response to pressure stimulus, and FAK is one of its downstream targets. Src phosphorylation due to PDGF receptor activation does not seem to be involved in the initial response. PMID- 11882599 TI - Lysophosphatidylcholine inhibits insulin-induced Akt activation through protein kinase C-alpha in vascular smooth muscle cells. AB - To better understand the intracellular signaling mechanism that causes the association of insulin resistance and hyperlipidemia with cardiovascular diseases, we specifically looked at the ability of lysophosphatidylcholine (lysoPC) to inhibit the Akt activation induced by insulin in cultured rat aortic vascular smooth muscle cells. LysoPC inhibited the insulin-induced phosphorylation of Akt at Ser473, and the inhibition was concentration dependent. Phorbol 12-myristate 13-acetate (PMA), a protein kinase C (PKC) activator, inhibited the insulin-induced phosphorylation of Akt. LysoPC stimulated PKC phosphorylation at Ser660, which was inhibited by the PKC inhibitor GF109203X. The PKC-alpha/beta-selective inhibitor Go6976 also blocked the PMA- and lysoPC induced inhibition of Akt phosphorylation by insulin. PKC-alpha, but not PKC beta, is expressed in vascular smooth muscle cells, and overexpression of PKC alpha, but not PKC-beta or PKC-delta, inhibited insulin-induced Akt activation. LysoPC rapidly stimulated PKC-alpha translocation to the membrane. In contrast, pretreatment with the p42/44 mitogen-activated protein kinase kinase inhibitor PD98059 or the p38 mitogen-activated protein kinase inhibitor SB203580 did not block the lysoPC-induced inhibition of Akt phosphorylation by insulin. In addition, lysoPC inhibited the insulin-induced tyrosine phosphorylation of insulin receptor substrate (IRS)-1 but not that of the insulin receptor beta subunit or insulin binding. PMA treatment or PKC-alpha overexpression also inhibited the tyrosine phosphorylation of IRS-1. From these data, we conclude that lysoPC negatively regulates the insulin signal at the point of IRS-1 through PKC-alpha in the vasculature, which may explain the association of hyperlipidemia with hyperinsulinemia in cardiovascular diseases. PMID- 11882600 TI - Excess norepinephrine impairs both endothelium-dependent and -independent vasodilation in patients with pheochromocytoma. AB - There is little information concerning the interaction of nitric oxide and norepinephrine (NE) on endothelial function in humans. The purpose of this study was to determine whether endothelial function is impaired by NE secreted from patients with pheochromocytoma (pheo) and whether surgical resection of the tumor improves endothelial function in these patients. We evaluated the forearm blood flow (FBF) response to acetylcholine (ACh), an endothelium-dependent vasodilator, and isosorbide dinitrate (ISDN), an endothelium-independent vasodilator, before and after adrenalectomy in 8 pheo patients, 20 normotensive subjects, and 20 patients with essential hypertension. FBF was measured using a mercury-filled silastic strain-gauge plethysmograph. The FBF response to ACh was the greatest in normotensive subjects and the least in pheo patients. The FBF response to ISDN was significantly less in pheo patients than in the other 2 groups, which had similar responses to ISDN. Adrenalectomy significantly decreased plasma and urinary NE, systolic and diastolic blood pressures, heart rate, and forearm vascular resistance. After adrenalectomy, FBF responses to both ACh and ISDN were enhanced in all pheo patients. The ratio of maximal ACh-stimulated FBF to maximal ISDN-stimulated FBF was significantly higher after adrenalectomy than before adrenalectomy (2.1 +/- 0.4 versus 1.1 +/- 0.1; P<0.05). The increase in maximal FBF response to ACh correlated significantly with the decrease in urinary excretion of NE (r=-0.62, P<0.01). These findings suggest that excess NE from pheo may predominantly impair endothelium-dependent vasodilation in humans. PMID- 11882601 TI - Downregulation of neuronal nitric oxide synthase and interleukin-1beta mediates angiotensin II-dependent stimulation of sympathetic nerve activity. AB - There is substantial evidence that angiotensin II (Ang II) enhances sympathetic nervous system (SNS) activity. We recently observed that nitric oxide and interleukin-1beta (IL-1beta) exert a tonic inhibitory action on central SNS activity. Moreover, in 2 rat models of neurogenic hypertension, one caused by intrarenal injection of phenol and the other by 5/6 nephrectomy, we observed that losartan, an Ang II type 1 receptor blocker, inhibits SNS activity and increases the abundance of IL-1beta and the neuronal isoform of nitric oxide synthase (nNOS) in the posterior hypothalamic nuclei (PH), paraventricular nuclei (PVN), and locus ceruleus (LC). This raises the possibility that the stimulatory effects of Ang II on central SNS activity may be mediated by inhibition of nNOS and IL 1beta. To test this hypothesis, we studied the effect of an intracerebroventricular (ICV) infusion of Ang II on blood pressure (BP), norepinephrine (NE) secretion from the PH, renal SNS activity (RSNA), and abundance of IL-1beta and nNOS mRNA in the PH, PVN, and LC of normal Sprague Dawley rats. Finally, we measured the concentration of nitrite/nitrate in the dialysate collected from the PH after Ang II or vehicle. ICV infusion of Ang II (100 ng/kg body wt dissolved in 10 microL of artificial cerebrospinal fluid) raised BP, RSNA, and NE secretion from the PH compared with control rats. Ang II reduced the abundance of IL-1beta and nNOS mRNA in the PH, PVN, and LC. Pretreatment with losartan (10 microg/kg body wt dissolved in 10 microL of aCSF) given ICV 20 minutes before Ang II abolished the effects of Ang II on BP, RSNA, and NE secretion from the PH and IL-1beta and nNOS mRNA. Ang II also decreased the secretion of NO from the PH. In conclusion, these studies suggest that Ang II inhibits the expression of IL-1beta and nNOS in the brain. Because locally produced NO exerts a tonic inhibitory action on SNS activity, the decrease in NO expression caused by Ang II results in greater SNS activity. PMID- 11882602 TI - Signaling mechanisms of heparin-binding epidermal growth factor-like growth factor in vascular smooth muscle cells. AB - A host of growth factors have been implicated in vascular pathologies; one such factor is heparin-binding epidermal growth factor-like growth factor (HB-EGF). Although HB-EGF has been shown to stimulate mitogenesis and chemotaxis of vascular smooth muscle cells (VSMC), its signaling mechanism remains undefined. In this study, we examined possible signal transduction pathways by which HB-EGF leads to mitogenesis in cultured rat VSMC. HB-EGF induced phosphorylation of the EGF receptor (EGFR) with maximum phosphorylation at 0.5 to 1 minute, whereas erbB4, the other receptor to which HB-EGF binds, was not activated on HB-EGF stimulation. HB-EGF induced a time- and concentration-dependent phosphorylation of mitogen-activated protein kinase (MAPK; p42/44 MAPK, extracellular signal regulating kinase [ERK] 1/2). It also activated Akt and p70S6 kinase (p70S6K) but not p38 MAPK. HB-EGF-induced phosphorylation of these kinases was blocked by the EGFR kinase inhibitor AG1478. To investigate signaling molecules involved in HB EGF-induced DNA synthesis, we pretreated VSMC with the specific ERK kinase mitogen-activated kinase (MEK) inhibitor PD98059 and the phosphatidylinositol 3 kinase inhibitor LY294002. These inhibitors significantly blocked HB-EGF-induced DNA synthesis. PD98059 inhibited HB-EGF-induced ERK activation, whereas it had no effect on Akt activation by HB-EGF. By contrast, LY294002 inhibited HB-EGF induced Akt and p70S6K activation without effecting ERK activation by HB-EGF. These results demonstrate that HB-EGF-induced mitogenesis requires both ERK and phosphatidylinositol 3-kinase (Akt and p70S6K) pathways activated through EGFR, thereby providing a new mechanistic insight by which HB-EGF contributes to vascular remodeling. PMID- 11882603 TI - A(2B) adenosine receptors stimulate growth of porcine and rat arterial endothelial cells. AB - The goal of this study was to determine which adenosine receptor subtype mediates growth stimulation by adenosine in arterial endothelial cells. In porcine coronary artery and rat aortic endothelial cells, 2-chloroadenosine (Cl-Ad), a metabolically stable analog of adenosine, stimulated DNA synthesis ((3)H thymidine incorporation), cellular proliferation (cell number), collagen synthesis ((3)H-proline incorporation), and cell migration. The growth effects of adenosine and Cl-Ad were mimicked by the adenosine receptor agonist 5'-N methylcarboxamidoadenosine but not by the adenosine receptor agonists N(6) cyclopentyladenosine, 4-aminobenzyl-5'-N-methylcarboxamidoadenosine or CGS21680, an agonist profile consistent with an A(2B) receptor-mediated effect. The adenosine receptor antagonists KF17837 and 1,3-dipropyl-8-p-sulfophenylxanthine but not 8-cyclopentyl-1,3-dipropylxanthine blocked the growth-stimulatory effects of Cl-Ad and 5'-N-methylcarboxamidoadenosine, an antagonist profile consistent with an A(2) receptor-mediated action. Treatment of endothelial cells with erythro-9-(2-hydroxy-3-nonyl) adenine plus iodotubericidin (inhibitors of adenosine deaminase and adenosine kinase, respectively) induced endothelial cell growth, and these effects were blocked by 1,3-dipropyl-8-p-sulfophenylxanthine and KF17837 but not 8-cyclopentyl-1,3-dipropylxanthine, suggesting that endothelial cell-derived adenosine induces growth via A(2) receptors. The growth stimulatory effects of Cl-Ad, 5'-N-methylcarboxamidoadenosine, and erythro-9-(2 hydroxy-3-nonyl) adenine plus iodotubericidin were abolished by antisense but not scrambled or sense oligonucleotides to the A(2B) receptor. Our findings strongly support the hypothesis that adenosine induces endothelial cell growth by activating A(2B) receptors. Thus, A(2B) receptors may play a critical role in regulating vascular remodeling associated with endothelial cell proliferation in angiogenesis, collateral vessel development, and recovery after vascular injury. Pharmacological or molecular biological activation of A(2B) receptors may be useful in modulating vascular remodeling. PMID- 11882604 TI - Novel receptors for ouabain: studies in adrenocortical cells and membranes. AB - Sodium-potassium pumps (Na pumps) are the only known plasma membrane receptors for cardiac glycosides. However, adrenocortical cells secrete an endogenous ouabain via an unknown mechanism that is subject to feedback inhibition via the cell surface. In addition, recent studies suggest that the induction of sustained hypertension by ouabain analogs in rats may be independent of Na pump inhibition. Accordingly, we used bovine adrenocortical cells and membranes to search for novel binding sites for ouabain. In high extracellular potassium solutions, the binding of ouabain to the Na pumps of cultured cells was suppressed, yet residual specific binding of (3)H-ouabain was observed. In high extracellular potassium, Scatchard analyses revealed a novel class of ouabain binding sites with high affinity (<50 nmol/L, < 2.5 x 10(5) sites/cell) that was distinct from the low affinity Na pump sites (>1 micromol/L, 4.5 x 10(6) sites/cell). Analysis of the kinetics for the dissociation of (3)H-ouabain from intact cells revealed components whose t(0.5) values were 6.5 minutes, 3.3 hours, and 33 hours and associated with novel sites, Na pumps, and lysosomal recycling, respectively. Studies with isolated membranes under ligand conditions where the participation of Na pumps was minimized revealed specific ouabain binding to novel sites that was saturable, time-dependent, of high affinity (K(d) approximately 15 nmol/L), and of low density (apparent B(max)=0.23 pmol/mg, c.f., Na pumps=10.2 pmol/mg). Ouabain binding to the novel sites was stimulated by high concentrations of KCl but was not affected by aldosterone or cortisol up to 30 micromol/L. Novel sites were not detected in skeletal muscle or liver membranes. Photoaffinity studies followed by SDS-PAGE showed ouabain-protectable labeling of membrane polypeptides with apparent molecular weights of 143, 113, and 65 kDa. We conclude that adrenocortical cells express ouabain receptors that are distinct from Na pumps. These novel receptors may be involved in the regulation and/or secretion of endogenous ouabain. PMID- 11882605 TI - Endothelin-1--induced enhancement of coronary smooth muscle contraction via MAPK dependent and MAPK-independent [Ca(2+)](i) sensitization pathways. AB - Endothelin-1 (ET-1) has been implicated in coronary vasospasm by enhancing coronary vasoconstriction to vasoactive eicosanoids, and a role for protein kinase C (PKC) activation has been suggested. However, the cellular mechanisms downstream from PKC activation are unclear. We investigated whether physiological concentrations of ET-1 enhance coronary smooth muscle contraction by activating a PKC-mediated signaling pathway involving tyrosine phosphorylation and activation of mitogen-activated protein kinase (MAPK). Cell contraction was measured in smooth muscle cells isolated from porcine coronary artery, [Ca(2+)](i) was measured in fura-2 loaded cells, and tissue fractions were examined for reactivity with anti-phosphotyrosine (P-Tyr) and anti-MAPK antibodies using immunoprecipitation and immunoblot analysis. In Hanks' solution (1 mmol/L Ca(2+)), ET-1 (10 pmol/L) did not increase basal [Ca(2+)](i) (81 +/- 2 nmol/L) but caused cell contraction (10%) that was inhibited by calphostin C (10(-6) mol/L), inhibitor of PKC, tyrphostin (10(-6) mol/L), inhibitor of tyrosine kinase, and PD098059 (10(-6) mol/L), inhibitor of MAPK kinase. The vasoactive eicosanoid prostaglandin F(2alpha) (PGF(2alpha); 10(-7) mol/L) caused increases in cell contraction (11%) and [Ca(2+)](i) (122 +/- 9 nmol/L) that were inhibited by the Ca(2+) channel blocker verapamil (10(-6) mol/L) but not by calphostin C, tyrphostin, or PD098059. Pretreatment with ET-1 for 10 minutes enhanced cell contraction to PGF(2alpha) (33%) with no additional increase in [Ca(2+)](i) (124 +/- 10 nmol/L). Activation of PKC by phorbol 12-myristate 13-acetate (PMA; 10(-7) mol/L) caused cell contraction and enhanced PGF(2alpha) contraction (32%) with no additional increase in [Ca(2+)](i) (126 +/- 9 nmol/L). The ET-1-- and PMA-induced enhancement of PGF(2alpha) contraction was abolished by verapamil or calphostin C but not by tyrphostin or PD098059. ET-1 and PMA caused significant increases in tyrosine phosphorylation of MAPK that were inhibited by calphostin C, tyrphostin, and PD098059. PGF(2alpha) did not cause any additional increases in tyrosine phosphorylation of MAPK in tissues untreated or pretreated with ET-1 or PMA. Thus, physiological concentrations of ET-1 activate a Ca(2+)-independent PKC mediated signaling pathway that involves tyrosine phosphorylation and activation of MAPK. The enhancement of PGF(2alpha)-induced coronary smooth muscle contraction by ET-1 involves additional activation of a Ca(2+)-sensitive PKC mediated pathway but not tyrosine phosphorylation or activation of MAPK. The MAPK dependent and MAPK-independent signaling pathways represent possible cellular mechanisms by which ET-1 could enhance coronary vasoconstriction to vasoactive eicosanoids in coronary vasospasm. PMID- 11882606 TI - Sustained activation of the central baroreceptor pathway in angiotensin hypertension. AB - Recent studies indicate that renal sympathetic nerve activity is chronically suppressed in angiotensin (Ang II) hypertension and that baroreflexes play a critical role in mediating this response. To support these findings, we determined whether the hypertension associated with chronic infusion of Ang II at 4.8 pmol/kg per minute (5ng/kg per minute) produces sustained activation of medullary neurons that participate in the central baroreceptor reflex pathway. We used Fos-like (Fos-Li) protein immunohistochemical methods to determine activation of neurons in the nucleus tractus solitarius (NTS), caudal ventrolateral medulla (CVLM), and rostral ventrolateral medulla (RVLM). Results were compared in three groups of chronically instrumented dogs subjected to infusion of: 1) saline (control); 2) Ang II-2 hours (acute); and 3) Ang II-5 days (chronic). Mean arterial pressure increased 22 +/- 3 and 35 +/- 3 mm Hg during acute and chronic Ang II infusion, respectively. There was little Fos-Li immunoreactivity in medullary neurons in control dogs. In contrast, during acute Ang II infusion there was a 2- to 3-fold increase in Fos-Li staining in the NTS and CVLM, but no increase in staining in RVLM neurons. As baroreceptor suppression of sympathoexcitatory cells in the RVLM is mediated by activation of neurons in the NTS and CVLM, these results were expected. More importantly, this same pattern of central neuronal activation was observed during chronic Ang II hypertension. Therefore, these results support recent findings indicating that baroreflex suppression of renal sympathetic nerve activity is a long-term compensatory response in Ang II hypertension. PMID- 11882607 TI - Brain amiloride-sensitive Phe-Met-Arg-Phe-NH(2)--gated Na(+) channels and Na(+) induced sympathoexcitation and hypertension. AB - Dietary and cerebrospinal fluid (CSF) Na(+) may act through brain amiloride sensitive, Phe-Met-Arg-Phe-NH(2) (FMRFamide)-gated Na(+) channels (FaNaChs) to cause sympathoexcitation and hypertension. We hypothesized that FaNaChs cause sympathoexcitation via the activation of brain "ouabain" and the brain renin angiotensin system. In conscious Wistar rats, intracerebroventricular (ICV) infusion of Na(+)-rich (0.3 mol/L) artificial CSF (aCSF) and ICV injection of angiotensin II or ouabain increase renal sympathetic nerve activity (RSNA), blood pressure (BP), and heart rate (HR). ICV benzamil, an amiloride analogue, did not affect baseline values and blocked the responses to ICV infusions of Na(+)-rich aCSF but not ICV angiotensin II or ouabain. ICV FMRFamide also increased RSNA, BP, and HR. Blocking brain "ouabain" with ICV antibody Fab fragments abolished the responses to both ICV FMRFamide and Na(+)-rich aCSF. In conscious spontaneously hypertensive rats (SHR) on a high salt intake for 6 weeks, prolonged ICV but not intravenous infusion of benzamil at 10 to 20 microg/h significantly decreased RSNA, BP, and HR in a dose-related manner. The extent of these responses was significantly smaller in SHR on regular salt intake. These findings suggest that benzamil-blockable brain FaNaChs represent the major mechanism through which a small increase in CSF Na(+) by ICV Na(+)-rich aCSF in Wistar rats or high salt intake in SHR initiates sympathoexcitation and hypertension. Enhanced Na(+) entry through FaNaChs appears to activate brain "ouabain" and the brain renin-angiotensin system and, thereby, increases the sympathetic outflow. Brain FaNaChs appear to contribute to the worsening of hypertension in SHR on a high salt diet and, to a small extent, to the maintenance of hypertension in SHR on a regular salt diet. PMID- 11882608 TI - Chronotropic action of angiotensin II in neurons via protein kinase C and CaMKII. AB - Angiotensin II (Ang II) plays an important role in the central control of blood pressure and baroreflexes. These effects are initiated by stimulation of Ang II type 1 (AT(1)) receptors on neurons within the hypothalamus and brain stem, and involve increasing the activity of noradrenergic, substance P, and glutamatergic pathways. The goal of this study is to investigate the intracellular signaling molecules, which are involved in mediating the Ang II-induced increases in neuronal activity. Using neurons in primary culture from newborn rat hypothalamus and brain stem, we have previously determined that Ang II elicits an AT(1) receptor-mediated inhibition of delayed rectifier K(+) current, a stimulation of Ca(2+) current, and a consequent increase in firing rate. In the present study we have demonstrated that this chronotropic action of Ang II in neuronal cultures involves activation of Ca(2+)-dependent signaling molecules. The Ang II-induced increase in firing rate was abolished by inhibition of phospholipase C with U73122 (10 micromol/L), and was attenuated by the protein kinase C inhibitor calphostin C (10 micromol/L) or by the calcium/calmodulin-dependent kinase II (CaMKII) inhibitor KN-93 (10 micromol/L). A combination of calphostin C and KN-93 completely inhibited this Ang II action. These results indicate that the AT(1) receptor-mediated increase in neuronal firing rate involves activation of both PKC and CaMKII, and suggest that these enzymes are potential targets for manipulating the central actions of Ang II. PMID- 11882609 TI - Obligatory role of protein kinase Cbeta and MARCKS in vesicular trafficking in living neurons. AB - Neurotransmitter release from neurons involves both vesicular trafficking and subsequent fusion of synaptic vesicles with the plasma membrane. The mechanisms involving the formation and fusion of vesicles that allow the exocytotic release of transmitters are understood well. Little is known, however, about the signaling mechanism involved in the trafficking of vesicles along the neurites. In this study, we used real-time confocal microscopy to search for evidence that vesicular trafficking in neurons requires the activation of protein kinase Cbeta (PKCbeta) and the myristoylated alanine-rich C kinase substrate (MARCKS) signaling pathway. Dopamine-beta-hydroxylase fused to green fluorescent protein has been used to trace vesicular movement. Angiotensin II, an established neuromodulatory hormone, stimulates translocation of green fluorescent protein dopamine-beta-hydroxylase vesicles from the cell body to neurites. This translocation was blocked by an antisense oligonucleotide to PKCbeta and MARCKS. Stimulation of PKC by other means, such as phorbol-12-myristate-13-acetate or carbachol, also resulted in the redistribution of fluorescence in a manner similar to that observed for angiotensin II. These observations demonstrate that PKCbeta-MARCKS signaling may be a general mechanism for the stimulation of vesicular trafficking in brain neurons. PMID- 11882610 TI - Prorenin-induced myocyte proliferation: no role for intracellular angiotensin II. AB - Cardiomyocytes bind, internalize, and activate prorenin, the inactive precursor of renin, via a mannose 6-phosphate receptor (M6PR)--dependent mechanism. M6PRs couple directly to G-proteins. To investigate whether prorenin binding to cardiomyocytes elicits a response, and if so, whether this response depends on angiotensin (Ang) II, we incubated neonatal rat cardiomyocytes with 2 nmol/L prorenin and/or 150 nmol/L angiotensinogen, with or without 10 mmol/L M6P, 1 micromol/L eprosartan, or 1 micromol/L PD123319 to block M6P and AT(1) and AT(2) receptors, respectively. Protein and DNA synthesis were studied by quantifying [(3)H]-leucine and [(3)H]-thymidine incorporation. For comparison, studies with 100 nmol/L Ang II were also performed. Neither prorenin alone, nor angiotensinogen alone, affected protein or DNA synthesis. Prorenin plus angiotensinogen increased [(3)H]-leucine incorporation (+21 +/- 5%, mean +/- SEM, P<0.01), [(3)H]-thymidine incorporation (+29 +/- 6%, P<0.01), and total cellular protein (+14 +/- 3%, P<0.01), whereas Ang II increased DNA synthesis only (+34 +/ 7%, P<0.01). Eprosartan, but not PD123319 or M6P, blocked the effects of prorenin plus angiotensinogen as well as the effects of Ang II. Medium Ang II levels during prorenin and angiotensinogen incubation were <1 nmol/L. In conclusion, prorenin binding to M6PRs on cardiomyocytes per se does not result in enhanced protein or DNA synthesis. However, through Ang II generation, prorenin is capable of inducing myocyte hypertrophy and proliferation. Because this generation occurs independently of M6PRs, it most likely depends on the catalytic activity of intact prorenin in the medium (because of temporal prosegment unfolding) rather than its intracellular activation. Taken together, our results do not support the concept of Ang II generation in cardiomyocytes following intracellular prorenin activation. PMID- 11882611 TI - Effect of TNF-alpha--converting enzyme inhibitor on insulin resistance in fructose-fed rats. AB - Insulin resistance is associated with hypertension, obesity, dyslipidemia, and type 2 diabetes. It is well known that tumor necrosis factor (TNF)-alpha is one of the factors linked to obesity-induced insulin resistance; however, there have been no reports on the role of TNF-alpha in insulin resistance in nonobese insulin-resistant hypertensives. We tested the hypothesis that TNF-alpha affects insulin resistance in nonobese insulin-resistant hypertensive fructose-fed rats (FFR) and that a TNF-alpha--converting enzyme (TACE) inhibitor that blocks TNF alpha secretion improves insulin resistance in FFR. Six-week-old male Sprague Dawley rats were fed either standard chow (control) or fructose-rich chow (FFR) for 6 weeks. For the last two weeks of a six-week period of either diet, the rats were treated with a vehicle (control or FFR) or a TACE inhibitor (100 mg/kg/d of KB-R7785; FFR+TACE-I) in peritoneal injection. At the age of 12 weeks, insulin sensitivity was assessed in all conscious rats by the euglycemic hyperinsulinemic glucose clamp technique. While FFR had higher blood pressure than the control rats (P<0.01), the TACE inhibitor did not change blood pressure. Insulin sensitivity (M-value) was reduced in FFR compared with that in the control rats (16.7 +/- 1.1 mg/kg per min and 10.3 +/- 0.6 mg/kg per min in the control rats and FFR, respectively, P<0.001), and the TACE inhibitor improved insulin sensitivity to the level of the control rats (14.3 +/- 1.2 mg/kg per min in FFR+TACE-I, P<0.01). These data indicate that TNF-alpha plays a major role in insulin resistance in nonobese insulin-resistant models and also suggest that TACE would be a good target for controlling insulin resistance not only in obese models but also in nonobese insulin-resistant models. PMID- 11882612 TI - Calcium-dependent synthesis of prostacyclin in ATP-stimulated venous endothelial cells. AB - Prostacyclin is a powerful vasodilator that is released from vascular endothelial cells. Previous studies in our laboratory have indicated that arachidonic acid metabolites from venous endothelium play an important role in the dilation of adjacent arterioles during muscle stimulation. Furthermore, recent studies have suggested that ATP released from red blood cells during hypoxia stimulates dilation of arterioles. We tested the hypothesis that an ATP-induced increase in intracellular Ca(2+) in venous endothelium promotes prostacyclin synthesis. Small branches of femoral veins were isolated from male golden hamsters, placed in a 1 mL bath, and cannulated for perfusion with 3-(N-morpholino) propanesulfonic acid (MOPS)-buffered physiological salt solution at 37 degrees C. Prostacyclin synthesis was determined by enzyme immunoassay of bath solution. Perfusion of veins with ATP increased prostacyclin synthesis from 50 +/- 5 to 627 +/- 46 pg/mL (n=49). ATP-induced prostacyclin synthesis was inhibited by removal of extracellular Ca(2+), chelation of intracellular Ca(2+) with 1,2-bis(2 aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (BAPTA) (10 micromol/L for 10 minutes), and preincubation with cytosolic phospholipase A(2) (PLA(2)) inhibitors, AACOCF(3), and bromoenol lactone. Changes in intracellular Ca(2+) in cultured human venous endothelial cells were assessed by fura-2 spectrofluorometry. ATP induced a transient Ca(2+) peak within seconds, and the subsequent Ca(2+) plateau was abolished by removal of extracellular Ca(2+). An increase in prostacyclin synthesis was detected in these cells 2 minutes after application of ATP. These findings suggest that the ATP-induced increase in intracellular Ca(2+) stimulates prostacyclin synthesis in venous endothelial cells. PMID- 11882613 TI - Inducible nitric oxide synthase inhibition attenuates renal hemodynamics during pregnancy. AB - Acute, nonselective nitric oxide synthase inhibition in the pregnant rat decreases glomerular filtration rate and renal plasma flow, suggesting a role for nitric oxide in mediating renal vasodilation during pregnancy. As mid-gestation in the rat is associated with a significant increase in renal protein expression of inducible nitric oxide synthase, the aim of this study was to examine the role of inducible nitric oxide synthase in mediating renal hemodynamics changes at mid gestation in the rat. At day 16 of pregnancy, glomerular filtration rate was significantly higher in pregnant rats compared with virgin rats (3.1 +/- 0.4 versus 2.7 +/- 0.3 mL/min, respectively; P<0.05), as was effective renal plasma flow (13.4 +/- 2.5 versus 10.9 +/- 2.2 mL/min, respectively; P<0.05). Acute administration of the inducible nitric oxide synthase selective inhibitor, AMT hydrochloride (750 nmol/h), markedly attenuated the increase in glomerular filtration rate observed in pregnant rats (2.3 +/- 0.2 mL/min, P<0.01 versus pregnant) without significantly altering glomerular filtration rate in virgin rats (2.1 +/- 0.2 mL/min). Acute AMT administration significantly decreased effective renal plasma flow in pregnant (8.9 +/- 1.8 mL/min, P<0.01 versus pregnant) and virgin rats (7.1 +/- 0.9 mL/min, P<0.05 versus virgin). Acute administration of EIT (380 nmol/h), another inducible nitric oxide synthase selective inhibitor, also attenuated pregnancy-induced increases in glomerular filtration rate (2.1 +/- 0.2, 2.8 +/- 0.3, and 2.3 +/- 0.3 mL/min; virgin, pregnant, and EIT, respectively) and effective renal plasma flow (8.5 +/- 1.1, 13.8 +/- 2.1, and 9.0 +/- 1.1 mL/min; virgin, pregnant, and EIT, respectively). Therefore, these findings suggest that inducible nitric oxide synthase may play an important role in mediating the renal hemodynamic changes that occur during normal pregnancy. PMID- 11882614 TI - Interaction of O(2)(-) and NO in the thick ascending limb. AB - Nitric oxide (NO) is an important regulator of NaCl absorption by the thick ascending limb of the loop of Henle (THAL). The free radical superoxide (O(2)(-)) reacts with NO, decreasing its bioavailability. O(2)(-) is produced by mitochondria and various oxidases, some of which are present in the THAL. However, the ability of the THAL to produce O(2)(-) and its interaction with NO have not been studied. We hypothesized that NO bioavailability is decreased by O(2)(-). THALs were isolated and perfused and NO production was measured with an NO-selective microelectrode. Addition of L-Arg (250 micromol/L), but not D arginine, to the bath increased NO release by 34.8 +/- 11.8 pA (n=7). The response to L-Arg was completely abolished by the NO synthase inhibitor L-NAME (n=7). Scavenging THAL O(2)(-) with the superoxide dismutase (SOD) mimetic Tempol (50 micromol/L) increased L-Arg-induced NO release. At all concentrations of L Arg tested (50, 100, 250, 500, and 750 micromol/L), further addition of Tempol to the bath significantly increased NO release by THALs. Addition of SOD (300 U/mL) to the bath increased L-Arg-induced NO levels by 82% (n=5; P<0.02). Pretreatment of THALs with the SOD inhibitor diethyl-dithiocarbamate (250 micromol/L) blunted L-Arg-induced NO release by 63% compared with untreated tubules (n=5; P<0.05). Finally, we tested the effect of Tempol on NO-induced inhibition of THAL chloride transport. Addition of L-Arg decreased THAL Cl(-) absorption by 35%. Subsequent addition of Tempol (50 micromol/L) to the bath further decreased Cl(-) absorption by 35% (n=6; P<0.05). We conclude that NO bioavailability in the THAL is decreased by O(2)(-). In addition, we believe our studies are the first to show that endogenous O(2)(-) may act as a physiological regulator of nephron NaCl transport. PMID- 11882615 TI - Altered nitric oxide synthase 3 distribution in mesenteric arteries of hypertensive rats. AB - A high-salt (HS) diet and angiotensin II (Ang II) are both associated with the development of hypertension and impaired endothelial function. We hypothesize that alterations in nitric oxide synthase (NOS) activity or subcellular localization of NOS 3 protein may contribute to endothelial dysfunction in salt dependent hypertension. To test this hypothesis, two models of salt-dependent hypertension were studied: DOCA-salt and Ang II. For Ang II hypertension, rats were divided into 4 groups: control on normal or HS diet, Ang II-infused on normal or HS diet. The mesenteric arterial bed was isolated and frozen in liquid nitrogen. Frozen arteries were homogenized and separated into cytosolic and particulate fractions. NOS activity was assayed by determining the conversion of (3)H-arginine to (3)H-citrulline in the absence and presence of the NOS inhibitor, Nomega-nitro-L-arginine. NOS 3 protein expression was significantly increased in the cytosol of arteries from DOCA-salt compared with placebo rats and in Ang II-infused and Ang HS rats compared with control. NOS 3 expression in the particulate fraction was comparable among all groups. NOS activity (pmol/30 min/total protein) was significantly increased in the cytosolic fraction of arteries from DOCA-salt rats compared with placebo and in Ang HS rats compared with control. NOS activity was comparable in the particulate fraction in all rat groups. In conclusion, there is an altered subcellular distribution of NOS 3 in salt-dependent hypertension that may contribute to the development of hypertension and endothelial dysfunction. PMID- 11882616 TI - Selective gene transfer to key cardiovascular regions of the brain: comparison of two viral vector systems. AB - The systemic renin-angiotensin system (RAS) plays a critical role in cardiovascular (CV) homeostasis. All components of the RAS are also known to be produced cell-specifically within specific brain regions, although the role of the brain RAS relative to the systemic RAS has remained a puzzle due to the difficulty of dissecting these two systems. Selectively targeting these regions with genes that modify the RAS could help unravel this puzzle. We compared the ability of adenovirus (Ad) and lentivirus (feline immunodeficiency virus, FIV) vectors to mediate gene delivery in vivo to the supraoptic nucleus (SON) and subfornical organ (SFO), two important CV control regions known to express the various RAS genes. SON or SFO of adult C57BL/6 mice (n=37) were stereotaxically injected with replication-deficient recombinant Ad or FIV harboring a beta galactosidase (beta-gal) reporter gene. At 1, 3, or 8 weeks post-injection, brain sections were processed for beta-Gal activity, double immunofluorescence to verify cell-type specificity of viral transduction, or immunohistochemical detection of inflammatory mediators. Our results demonstrate that: (1) murine SFO and SON can be selectively targeted for gene transfer in vivo;(2) FIV mediated neuron-specific gene delivery, whereas Ad transduced both neuronal and glial cell types in SFO and SON; (3) Ad injected into the SON transduced neurons within the SFO through retrograde transport, whereas FIV did not; (4) beta-gal activity remained stable for 3 weeks but then declined by 8 weeks with Ad, while minimal decline occurred with FIV; (5) FIV did not cause inflammatory responses, whereas infiltrate was detectable in Ad-injected SFO and SON. These vectors are potentially important tools for dissecting the cell- and site-specific components of the brain RAS and other important CV regulatory systems within this circuitry, and may have therapeutic applications for centrally mediated CV diseases. PMID- 11882617 TI - Cytochrome P450-dependent eicosapentaenoic acid metabolites are novel BK channel activators. AB - P450-dependent arachidonic acid (AA) metabolites regulate arterial tone by modulating calcium-activated (BK) potassium channels in vascular smooth muscle cells (VSMC). Because eicosapentaenoic acid (EPA) has been reported to improve vascular function, we tested the hypothesis that P450-dependent epoxygenation of EPA produces alternative vasoactive compounds. We synthesized the 5 regioisomeric epoxyeicosattrienoic acids (EETeTr) and examined them for effects on K(+) currents in rat cerebral artery VSMCs with the patch-clamp technique. 11(R),12(S) epoxyeicosatrienoic acid (50 nmol/L) was used for comparison and stimulated K(+) currents 6-fold at +60 mV. However, 17(R),18(S)-EETeTr elicited a more than 14 fold increase. 17(S),18(R)-EET and the remaining four regioisomers were inactive. The effect of 17(R),18(S)-EETeTr was blocked by tetraethylammonium but not by 4 aminopyridine. VSMCs expressed P450s 4A1 and 4A3. Recombinant P450 4A1 hydroxylated EPA at C-19 and C-20 and epoxygenated the 17,18-double bond, yielding the R, S- and S, R-enantiomers in a ratio of 64:36. We conclude that 17(R),18(S)-EETeTr represents a novel, potent activator of BK potassium channels. Furthermore, this metabolite can be directly produced in VSMCs. We suggest that 17(R),18(S)-EETeTr may function as an important hyperpolarizing factor, particularly with EPA-rich diets. PMID- 11882618 TI - Cardiac damage prevention by eplerenone: comparison with low sodium diet or potassium loading. AB - To determine the extent to which dietary sodium modulates aldosterone-induced cardiovascular damage, and to determine whether increased dietary potassium can prevent this damage, we used the Nomega-nitro-L-arginine methyl ester (L NAME)/angiotensin II (Ang II) rat model of cardiac injury. This model is dependent on the presence of aldosterone for the occurrence of myocardial damage. Two sets of experiments were performed. In the first set, the following groups were studied: (1) 1% NaCl to drink (control group); (2) L-NAME/Ang II with water to drink (low salt group); (3) L-NAME/Ang II/1% NaCl (high salt group); (4) L NAME/Ang II/1% NaCl/eplerenone (eplerenone group). Systolic blood pressure increased similarly in all groups compared with controls. Compared with the controls, the high salt group, but not the low salt or eplerenone groups, developed significant myocardial damage. In the second set of experiments three groups of animals were studied: (1) L-NAME/Ang II/1%NaCl (high salt group) (2) L NAME/Ang II/1%NaCl/eplerenone (eplerenone group), and (3) L-NAME/Ang II/1%NaCl with an extra 1% KCl in food (high dietary potassium group). Eplerenone, but not dietary potassium supplementation, prevented the development of cardiac damage. Thus, mineralocorticoid receptor antagonist treatment and low sodium diet were effective in preventing cardiac damage, which suggests that a minimal level of aldosterone and a moderately high sodium diet are both required for the development of the cardiovascular damage in the L-NAME/Ang II model. The inability of potassium supplementation to reduce myocardial damage suggests that eplerenone's protective effect is not due to its potassium-sparing ability, but is rather related to some other feature of its selective aldosterone antagonism. PMID- 11882619 TI - Neprilysin inhibitors potentiate effects of bradykinin on b2 receptor. AB - Some beneficial effects of angiotensin-I--converting enzyme (ACE, kininase II) inhibitor therapy are attributed to enhancing the activity of bradykinin on its B(2) receptor. Independent of inhibition of bradykinin hydrolysis, ACE inhibitors enhance the action of bradykinin on its B(2) receptor by inducing crosstalk between ACE and the receptor. We investigated whether inhibitors of another kininase II-type enzyme, neprilysin (neutral endopeptidase 24.11; NEP), could augment bradykinin effects unrelated to blocking its breakdown using a NEP resistant bradykinin analog as ligand. We used transfected Chinese hamster ovary (CHO) cells stably expressing human B(2) receptor and NEP (CHO/NEP-B(2)) or only B(2) (CHO/B(2)) as control and human pulmonary fibroblasts (IMR90), expressing B(2), but more NEP than ACE. NEP inhibitor phosphoramidon (100 nmol/L), or omapatrilat, which inhibits both NEP and ACE, did not potentiate bradykinin in CHO/B(2) cells. In IMR90 cells, 10 nmol/L bradykinin elevated [Ca(2+)](i) and desensitized the receptor. Adding either 100 nmol/L omapatrilat or phosphoramidon resensitized the receptor to the ligand, which was abolished by receptor blocker HOE 140. Arachidonic acid release by bradykinin from CHO/NEP-B(2) cells was also augmented by 100 nmol/L phosphoramidon or omapatrilat about 3-fold, and again, the inhibitors resensitized the desensitized B(2) receptor. The inhibitors did not potentiate bradykinin when soluble rNEP was added to the medium of CHO/B(2) cells. Similar to ACE, NEP inhibitors potentiated bradykinin independent of inhibiting inactivation. Consequently, omapatrilat could augment bradykinin effects on B(2), when either ACE or NEP is expressed close to receptor on cell membrane. PMID- 11882620 TI - Mechanism by which superoxide potentiates tubuloglomerular feedback. AB - The macula densa detects changes in NaCl concentration in tubular fluid and transmits a feedback signal, known as tubuloglomerular feedback (TGF), which helps to control glomerular afferent arteriole resistance. We and other investigators have reported that synthesis of NO in the macula densa inhibits TGF. NO can be scavenged by superoxide (O(-)(2)) to form peroxynitrite, effectively reducing the bioavailability of NO; there is growing evidence that O( )(2) regulates vascular tone in the kidney. We hypothesized that O(-)(2) produced in the macula densa enhances TGF and this effect acts only in an autocrine manner within the cells of the macula densa. Afferent arterioles and attached macula densas from Sprague-Dawley rats were simultaneously microperfused in vitro and TGF response examined before and after perfusing the tubular lumen, bath, or vascular lumen with a superoxide scavenger. The macula densa was perfused with solutions containing either 5 mmol/L Na(+) and 3 mmol/L Cl(-) (low NaCl) or 80 mmol/L Na(+) and 77 mmol/L Cl(-) (high NaCl) while keeping pressure in the afferent arteriole constant at 60 mm Hg. When 10(-4) M Tempol, a stable membrane permeant superoxide dismutase (SOD) mimetic, was added to the tubular lumen, it inhibited TGF by 56% (before Tempol: TGF, 3.2 +/- 0.3 microm; after Tempol: TGF, 1.4 +/- 0.2 microm; n=6; P<0.05, control versus Tempol). Adding Tempol to the bath inhibited TGF by 48% (before Tempol: TGF, 2.5 +/- 0.25 microm; after Tempol: TGF, 1.3 +/- 0.18 microm; n=6; P<0.05). However, adding Tempol to the vessel lumen did not change TGF response significantly (before Tempol: TGF, 2.7 +/- 0.37 microm; after Tempol: TGF, 3.2 +/- 0.25 microm; n=7; P=0.25). When 300 U/mL of the enzyme SOD, which is not membrane-permeant, was added to either the tubular lumen or bath, it had no effect on TGF response. Finally, to determine whether the effect of O(-)(2) in the macula densa is mediated by its scavenging of NO, 7 nitroindazole (7-NI) was added to the macula densa to inhibit neuronal nitric oxide synthase (nNOS). In the presence of 7-NI, Tempol had no effect (7-NI only: TGF, 3.0 +/- 0.4 microm; 7-NI plus Tempol: TGF, 2.8 +/- 0.5 microm; n=6; P=0.343). Our findings suggest that (1) reducing O(-)(2) increases the bioavailability of NO, which inhibits TGF, (2) both O(-)(2) and NO act within the macula densa, and (3) O(-)(2) appears to have no effect on its own. PMID- 11882621 TI - Lower blood pressure in floxed angiotensinogen mice after adenoviral delivery of Cre-recombinase. AB - Recent experimental evidence suggests a role for tissue renin-angiotensin systems in the development of hypertension. To test the importance of tissue renin angiotensin systems in the development and maintenance of angiotensin II dependent hypertension, we generated a transgenic model in which exon 2 of the human angiotensinogen gene is flanked by loxP sites (hAGT(flox)) so that this region of the gene can be deleted by the cre-recombinase. Double transgenic human renin and hAGT(flox) (R(+)/A(+flox)) mice of two independent lines exhibited elevated blood pressure. Acute administration of an adenovirus containing cre recombinase (Adcre) lowered blood pressure by 30 mm Hg over a 4-day period as measured with fluid filled catheters. The chronic effect of Adcre administration on blood pressure was determined by radiotelemetry in a separate group of R(+)/A(+flox) mice. Blood pressure decreased by 25 mm Hg from baseline by day 8 post-Adcre, but increased on each day thereafter until it was 90% of baseline by day 21 post-Adcre. Expression analysis indicated the absence of detectable hAGT mRNA in the liver at day 5 post-Adcre, but reappeared at normal levels at days 14 to 21 post-Adcre. These studies suggest that Adcre is effective for acute, but not chronic, elimination of hepatic hAGT. Chronic elimination of hepatic hAGT will likely require the use of transgenic mice endogenously expressing cre recombinase in the liver. PMID- 11882622 TI - Perindopril alters vascular angiotensin-converting enzyme, AT(1) receptor, and nitric oxide synthase expression in patients with coronary heart disease. AB - Angiotensin-converting enzyme inhibitors (ACEi) reduce cardiovascular morbidity and mortality by improving coronary perfusion, reducing ventricular hypertrophy and remodeling, and preventing progression of coronary atherosclerosis. However, the cellular mechanisms underlying the beneficial effects of ACEi are not fully understood. We studied the in vivo effects of ACE inhibition with perindopril on cellular expression of ACE, AT(1) receptors and 2 nitric oxide synthase (NOS) isoforms, endothelial (eNOS) and inducible NOS (iNOS), in human blood vessels using quantitative in vitro autoradiography and immunocytochemistry. Seven patients with ischemic heart disease were treated with perindopril (4 mg/d) for up to 5 weeks before elective coronary bypass surgery, whereas controls did not receive the ACEi (n=7). Perindopril decreased plasma ACE by 70% and the plasma angiotensin II to angiotensin I ratio by 57% and reduced vascular ACE to approximately 65% of control levels in both endothelium and adventitia. By contrast, AT(1) receptor binding in vascular smooth muscle cells was increased by 80% in patients treated with perindopril as confirmed by immunocytochemistry. eNOS was expressed primarily in endothelial cells, whereas little iNOS expression occurred in vascular smooth muscle cells of untreated patients. Both eNOS and iNOS expression seemed to increase during perindopril treatment. These results suggest that suppression of angiotensin II formation in the vascular wall and increased expression of eNOS and iNOS during ACE inhibition may be beneficial in reversing endothelial dysfunction in patients with cardiovascular disease. Because vascular AT(1) receptor expression is increased during chronic ACE inhibition, more clinical studies are required to determine whether it is necessary to combine ACE inhibitors and AT(1) receptor antagonists in clinical management of heart failure, coronary heart disease, and hypertension PMID- 11882623 TI - Regulation of cyclooxygenase- and cytochrome p450-derived eicosanoids by heme oxygenase in the rat kidney. AB - Heme oxygenase enzymes (HO-1 and HO-2) catalyze the conversion of heme to biliverdin, free iron, and carbon monoxide (CO). Heme and products derived from its metabolism potentially influence renal function and blood pressure by affecting the expression and/or activity of hemeproteins, including cytochrome P450 (CYP4A) monooxygenases and cyclooxygenases (COX-1 and COX-2). We studied HO isoform expression and examined the effect of HO-1 induction by SnCl(2) on CYP4A and COX expression and activity in the rat kidney. HO-1 protein levels in kidney tissues from untreated rats were barely detectable, whereas HO-2 protein was expressed in all kidney structures examined and its levels were higher in the outer medulla followed by the inner medulla/papilla and cortex. HO-2 expression along the nephron followed its regional distribution, ie, the highest levels were detected in the medullary thick ascending limb (mTAL) and inner medullary collecting ducts followed by proximal tubules. SnCl(2) Treatment did not significantly affect HO-2 expression or distribution; however, it markedly increased HO-1 protein in the inner and outer medulla, specifically, in the inner medullary collecting ducts and mTAL. CYP4A expression and 20 hydroxyeicosatetraenoic acid (20-HETE) synthesis were the highest in the outer medulla followed by the cortex and inner medulla/papilla. SnCl(2) treatment reduced cortical and inner medullary CYP4A protein levels by 60% and 50% and inhibited 20-HETE synthesis by 90% and 60%, respectively. Despite a significant induction of HO-1 protein in the outer medulla, CYP4A expression and 20-HETE synthesis were hardly affected. SnCl(2) treatment did not affect COX-1 expression but markedly reduced cortical and medullary COX-2 protein levels. We conclude that HO isoform expression is segmented within the kidney and along the nephron and that treatment with an HO-1 inducer suppressed the levels of CYP4A and COX-2 proteins in a tissue-specific manner with concomitant effects on their activity. Such interactions may play an important role in the regulation of renal function. PMID- 11882624 TI - Native LDL induces proliferation of human vascular smooth muscle cells via redox mediated activation of ERK 1/2 mitogen-activated protein kinases. AB - This study investigated mechanisms underlying native low-density lipoprotein (LDL)-stimulated proliferation of human vascular smooth muscle cells (VSMC). Experiments were performed to determine whether native LDL affects reactive oxygen species (ROS) formation and activity of extracellular signal-regulated kinase 1/2 (ERK1/2), and whether redox-sensitive pathways contribute to LDL induced cell proliferation. Native LDL (100 microg/mL, 24 hours) increased cell proliferation (to 303 to 388% of control, P<0.0001) as determined by [methyl (3)H] thymidine incorporation. This effect was completely blocked either by the antioxidants N-acetylcysteine, Tiron, or nordihydroguaiaretic acid; the flavin inhibitor diphenylene iodonium; or superoxide dismutase (all P<0.0001), and partly blocked by ERK-inhibitor PD98059 or meclofenamate (P<0.01). Exposure of VSMC to native LDL for 20 minutes stimulated ROS formation, measured by dichlorodihydrofluorescein oxidation, and increased ERK1/2 activity by 3.1-fold (P<0.001). The latter effect was sensitive to MEK1/2 inhibitor PD98059 and Tiron (P<0.001), and in part to N-acetylcysteine or diphenylene iodonium (P<0.05). These results demonstrate that native LDL induces acute formation of ROS and subsequent activation of redox-sensitive ERK 1/2 mitogen-activated protein kinases, pathways that appear to be important for mitogenic signaling of native LDL in human vascular smooth muscle cells. PMID- 11882625 TI - Vigilant vector: heart-specific promoter in an adeno-associated virus vector for cardioprotection. AB - Repeated bouts of ischemia in the heart lead to fibrosis and eventually to heart failure. Although certain genes, such as SOD or hemoxygenase and antisense to AT(1)R, ACE, and (beta(1)-AR can provide short-term protection of the heart from ischemia, there is no known mechanism for constantly responding to repeated incidences of ischemia. We hypothesized that a "vigilant vector," designed to be expressed specifically in the heart and switch on therapeutic genes only during hypoxia, would provide cardioprotection. To attain cardiac specificity, we inserted an MLC2v promoter into an adeno-associated virus (AAV) designed to deliver AS to AT(1)R and gfp. In in vitro experiments in cardiomyocytes (H9C2 cells), the MLC2v-AAV-gfp drove gene expression in all cells at levels comparable to a cytomegalovirus (CMV) promoter. In in vivo experiments, the rAAV-MLC2v-gfp was injected intravenously into mice or rats. Green fluorescence protein (GFP) DNA was located in kidney, heart (right and left ventricle), lung, adrenal and spleen. GFP mRNA, however, was expressed only in the heart and absent in other tissues. To switch on the rAAV transgene during ischemia, we inserted a hypoxia response element (HRE). This upregulates transcription when O(2) levels are low. Thus, there are 4 components to the vigilant vector; a gene switch (HRE), a heart specific promoter (MLC2v), a therapeutic gene (AS-AT(1)R) and a reporter gene (gfp). To silence or lower basal level of expression while retaining specificity, we have reduced the length of the MLC2v promoter from 3 kb to 1775 bp or 281 bp. The truncated promoter is equally effective in heart specific expression. Preliminary studies with the rAAV-HRE-gfp in vitro show an increased expression in 1% O(2) in 4 to 6 hours. By adding additional hypoxia-inducible factor (HIFalpha) (5 microg), the MLC2v-gfp expression is increased by 4-fold in 1% O(2). Further amplification of the gene to 400-fold in 1% O(2) can be achieved with a double plasmid. The construct may serve as a prototype "vigilant vector" to switch on therapeutic genes in specific tissue with physiological signals. PMID- 11882626 TI - Sympathetic activation in adipose tissue and skeletal muscle of hypertensive rats. AB - The activation of the sympathetic nervous system is a common feature of arterial hypertension and other cardiovascular diseases. This activation might be dependent on an altered baroreflex control of vascular resistance of which the inhibitory response on sympathetic activity appears impaired. The aim of the study was to monitor during the natural course of arterial hypertension in spontaneously hypertensive (SHR) and age-matched Wistar Kyoto (WKY) rats (5, 16, 30, and 54 weeks of age) the peripheral sympathetic activity expressed as interstitial norepinephrine (NE) release and as tyrosine hydroxylase (TH) activity, the rate-limiting enzyme of NE synthesis, in the differently baroreflex controlled subcutaneous adipose tissues and skeletal muscles. Blood pressure and plasma NE in SHR were similar to WKY at 5 weeks of age but increased at all other ages. Body weight was similar in both 5-week-old rats but reduced in SHR at all other ages. The interstitial NE levels were greater in both SHR tissues at all ages as compared with WKY. In adipose tissue of SHR, TH activity was higher at all ages as compared with WKY, whereas TH activity in skeletal muscle was higher only after the development of hypertension. These data show that in both SHR tissues, an increase of interstitial NE release is always present during its lifespan. This suggests that increased sympathetic activation in the SHR model is not specific to baroreflex-controlled tissues such as skeletal muscle but involves also subcutaneous adipose tissue, the sympathetic efferents of which are independent from baroreflexes. PMID- 11882627 TI - Nitric oxide produced by THAL nitric oxide synthase inhibits TGF. AB - Nitric oxide (NO) produced by neuronal NO synthase (nNOS) in the macula densa decreases tubuloglomerular feedback (TGF). NO produced by NOS in the thick ascending limb (THAL) inhibits NaCl transport. We hypothesized that NO produced by NOS in the THAL reaches the macula densa and inhibits TGF. Rabbit afferent arterioles and attached macula densa were simultaneously microperfused in vitro. TGF response was determined by measuring afferent arteriole diameter before and after increasing NaCl in the macula densa perfusate. When the nNOS inhibitor 7 nitroindazole (7-NI) (10 micromol/L) was added to the macula densa lumen, it increased TGF from 2.3 +/- 0.2 to 3.5 +/- 0.5 microm (P<0.02; n=6). In the presence of 7-NI, N(omega)-nitro-L-arginine methyl ester (L-NAME) (1 mmol/L) enhanced TGF from 2.6 +/- 0.3 to 4.0 +/- 0.5 microm (P<0.02; n=6) when the macula densa was perfused orthograde via the THAL, whereas it had no effect on TGF when the macula densa was perfused retrograde via the distal tubule (DT). Inhibition of macula densa soluble guanylate cyclase with LY83583 (1 micromol/L) blocked the effect of NO produced by THAL NOS when the macula densa was perfused via the THAL. We concluded that NO produced by THAL NOS acts as a paracrine factor, reaching the macula densa and inhibiting TGF. PMID- 11882628 TI - Increased renal medullary oxidative stress produces hypertension. AB - The present study examined whether chronic increased oxidative stress within the medulla of the kidney lowers medullary blood flow and leads to hypertension. Optical fibers were implanted into the renal cortex and medulla of uninephrectomized Sprague-Dawley rats (Harlan Sprague-Dawley, Madison, Wis) for the daily measurement of blood flow to these regions using laser-Doppler flowmetry techniques, while arterial pressure was measured from an indwelling aortic catheter. A renal medullary interstitial catheter was implanted for the continuous delivery of the superoxide dismutase (SOD) inhibitor, diethyldithiocarbamic acid (DETC), at a dose of 7.5 mg/kg/d. Renal interstitial superoxide (O(2)(-)) levels were determined by perfusing an O(2)(-) sensitive fluorescent dye, dihydroethidium, through a microdialysis probe implanted into the medulla. Urine samples (24 hours) were collected for measurements of isoprostane excretion. The results indicate that medullary DETC infusions increased tissue O(2)(-) concentrations in the renal medulla (93.4 +/- 22.3,n=8, saline and 867.3 +/- 260.2, n=8, DETC; fluorescence units) and increased urinary 8-isoprostane excretion (4.1 +/- 0.4 ng/d, n=9, saline and 8.8 +/- 1.6 ng/d, n=10, DETC). Mean arterial pressure increased 24 hours after the start of intrarenal DETC infusion and remained nearly 20 mm Hg above control pressure throughout the 5 days of medullary SOD inhibition. During chronic medullary DETC infusion, medullary blood flow was significantly reduced (42.7%), whereas cortical blood flow was unchanged. Intravenous infusion of the same dose of DETC produced no changes in renal medullary or cortical blood flow or arterial blood pressure. The present experiments indicate that an increase in superoxide concentration within the renal medulla selectively reduces medullary blood flow resulting in chronic hypertension. PMID- 11882629 TI - Function and regulation of endothelin-1 and its receptors in salt sensitive hypertension induced by sensory nerve degeneration. AB - To determine the role of endothelin-1 (ET-1) and its receptors in salt-sensitive hypertension induced by sensory nerve degeneration, selective ET(A) antagonist (ABT-627) and ET(B) antagonist (A-192621) were used. Newborn Wistar rats were given vehicle or 50 mg/kg capsaicin subcutaneously on the first and second days of life. After the weaning period, male rats were divided into eight groups, and subjected to the following treatments for 2 weeks: control + normal salt diet (Con+NS, 0.5%), control + high salt diet (Con+HS, 4%), control + high salt diet + ABT-627 (Con+HS+ABT-627), control + high salt diet + A-192621 (Con+HS+A-192621), capsaicin + normal salt diet (Cap+NS), capsaicin + high salt diet (Cap+HS), capsaicin + high salt diet + ABT-627 (Cap+HS+ABT-627), capsaicin + high salt diet + A-192621 (Cap+HS+A-192621). Both ABT-627 (5 mg/kg/d) and A-192621 (30 mg/kg/d) were given by oral gavage twice a day. Mean arterial pressure (MAP, mm Hg) was higher in Con+HS+A-192621 (141 +/-11) than in Con+NS (94 +/- 10), Con+HS (95 +/- 5), and Con+HS+ABT-627 (97 +/- 6) (P<0.05). MAP was also higher in Cap+HS (152 +/ 6) and Cap+HS+A-192621 (180 +/- 7) than in Cap+NS (99 +/- 3) and Cap+HS+ABT-627 (104 +/- 5) (P<0.05), and it was higher in Cap+HS+A-192621 than in Cap+HS (P<0.05). Enzyme immunometric assay showed that ET-1 plasma concentration (pg/mL) was higher in Con+HS+A-192621 (7.59 +/- 0.78) than in Con+NS (2.68 +/- 0.56), Con+HS (2.50 +/- 0.92), and Con+HS+ABT-627 (3.54 +/- 0.79) (P<0.05). ET-1 plasma concentration was also higher in Cap+HS (8.95 +/-.16), Cap+HS+ABT-627 (9.82 +/- 1.22) and Cap+HS+A-192621 (10.97 +/- 0.57) than in Cap+NS (3.06 +/- 0.73) (P<0.05). We conclude that blockade of the ET(A) receptor prevents the development of salt sensitive hypertension induced by sensory nerve degeneration, indicating that activation of the ET(A) receptor by increased plasma ET-1 level contributes to elevation of blood pressure in this model. In contrast, blockade of the ET(B) receptor leads to an increase in blood pressure in both normal and sensory nerve degenerated rats fed a high salt diet. These results suggest that ET(B) plays an antihypertensive role in response to high salt intake under both normal and sensory nerve degenerated conditions. PMID- 11882630 TI - Fibrosis, matrix metalloproteinases, and inflammation in the heart of DOCA-salt hypertensive rats: role of ET(A) receptors. AB - In deoxycorticosterone acetate (DOCA)-salt hypertension, the endothelin-1 system is activated and plays a role in cardiac fibrosis. Remodeling of extracellular matrix (ECM) may lead to interstitial fibrosis, which may contribute to heart failure. Imbalance in synthesis and degradation of the ECM by matrix metalloproteinases (MMPs) as well as inflammation may play a role in matrix protein deposition and cardiac remodeling in hypertension. We measured expression of the extracellular matrix protein fibronectin, the activity of the gelatinases MMP-2 and MMP-9, the proinflammatory transcription factor NFkappaB, and the adhesion molecules, vascular cell adhesion molecule (VCAM)-1 and platelet endothelial cell adhesion molecule (PECAM)-1 in hearts of DOCA-salt hypertensive (DS) rats treated or not with the endothelin ET(A) antagonist BMS 182874 (BMS). Unilaterally nephrectomized rats (UniNx) were compared with DS rats treated or not with BMS 40 mg/kg/d. Fibronectin deposition was detectable at the first week, and remained elevated thereafter. This increase was abrogated by administration of the ET(A) antagonist. Enzymatic activity of gelatinases was increased (P<0.01) in DS compared with control during the first and second week. BMS blocked the increase of MMP-2 and MMP-9 activity at week 1 (P<0.05); MMP activity remained lower than in DS at week 2. NF-kappaB binding activity in DS was higher (P<0.05) than it was in controls during the second week, and was reduced by BMS. The adhesion molecules VCAM-1 and PECAM-1, and the antiapoptotic molecule xIAP were upregulated in the left ventricle of the heart of DS rats and downregulated in the rats treated with the ET(A) antagonist. In conclusion, cardiac extracellular remodeling in rats with endothelin-dependent hypertension was associated with increased fibronectin, MMP activity, and upregulation of inflammatory mediators, all of which were reduced by ET(A) antagonism. PMID- 11882631 TI - Lacidipine inhibits adhesion molecule and oxidase expression independent of blood pressure reduction in angiotensin-induced vascular injury. AB - Dihydropyridines can inhibit gene expression in-vitro and may have a protective vascular effect independent of blood pressure reduction. We tested the hypothesis that lacidipine prevents induction of inducible NO synthase (iNOS), influences leukocyte adhesion and infiltration, inhibits nuclear factor (NF)-kappaB transcription factor activity, and ameliorates end-organ damage in a transgenic rat model of angiotensin (Ang) II--dependent organ sclerosis. We treated rats transgenic for human renin and angiotensinogen (dTGR) from week 4 to 7 with lacidipine (0.3 or 3 mg/kg by gavage). Blood pressure was measured by tail cuff. Organ damage was assessed by histology and immunohistochemistry. Adhesion molecules and cytokines were analyzed by immunohistochemistry. Transcription factors were analyzed by mobility shift assays. Untreated dTGR developed moderate hypertension, cardiac hypertrophy, and severe renal damage with albuminuria. Lacidipine decreased blood pressure slightly at the low dose and substantially at the higher dose. However, both treatments reduced albuminuria and plasma creatinine to the same degree (P<0.05). Intercellular adhesion molecule-1 (ICAM 1) was markedly reduced by lacidipine as well as renal neutrophil and monocyte infiltration. Lacidipine reduced mitogen-activated protein (MAP) kinase phosphorylation and iNOS expression in both cortex and medulla. NF-kappaB and AP 1 were activated in dTGR but reduced by lacidipine. Lacidipine ameliorates Ang II induced end-organ damage independent of blood pressure lowering, perhaps by inhibiting the MAP kinase pathway and NF-kappaB activation. PMID- 11882632 TI - Soluble epoxide hydrolase inhibition lowers arterial blood pressure in angiotensin II hypertension. AB - Epoxyeicosatrienoic acids (EETs) have antihypertensive properties and play a part in the maintenance of renal microvascular function. A novel approach to increase EET levels is to inhibit epoxide hydrolase enzymes that are responsible for conversion of biologically active EETs to dihydroxyeicosatrienoic acids (DHETs) that are void of effects on the preglomerular vasculature. We hypothesized that inhibition of soluble epoxide hydrolase (sEH) would lower blood pressure in angiotensin II (Ang II) hypertension. Rat renal cortical tissue was harvested and urine collected 2 weeks following implantation of an osmotic minipump containing Ang II (60 ng/min). Renal cortical sEH protein expression was significantly higher in Ang II hypertension compared with normotensive animals. Likewise, urinary 14,15-DHET levels were significantly increased in hypertensive compared with normotensive animals and averaged 8.1 +/- 1.3 and 2.7 +/- 1.1 ng/d; respectively. In additional experiments, the sEH inhibitor N-cyclohexyl-N-dodecyl urea (NCND; 3 mg/d) or vehicle (corn oil, 0.5 mL) was administered daily by intraperitoneal injection starting on day 10. Administration of NCND for 4 days lowered systolic blood pressure by 30 mm Hg in Ang II hypertensive animals, whereas the corn oil vehicle had no effect on blood pressure in normotensive or Ang II hypertensive animals. Measurement of blood pressure by indwelling arterial catheters in conscious animals with free movement in their cages confirmed that NCND had antihypertensive properties. Arterial blood pressure averaged 119 +/- 5 mm Hg in normotensive, 170 +/- 3 mm Hg in hypertensive and 149 +/- 10 mm Hg in NCND-treated, Ang II-infused animals. Administration of the potential metabolite of NCND, N-cyclohexylformamide to Ang II hypertensive rats did not lower the systolic blood pressure. These studies demonstrate that increased sEH expression in the Ang II hypertensive kidney leads to increased EET hydration. Moreover, sEH plays a role in the regulation of blood pressure, and inhibition of sEH during Ang II hypertension is antihypertensive. PMID- 11882633 TI - Hypoxia inducible double plasmid system for myocardial ischemia gene therapy. AB - Coronary artery disease frequently involves repeated bouts of myocardial ischemia. To automatically up-regulate the cardioprotective transgenes under hypoxic ischemia, a "vigilant vector" gene therapy system was developed and tested in a rat embryonic myocardial cell line (H9c2). In the vigilant vector, a hypoxia response element-incorporated promoter was used as a switch to turn on the gene expression in response to hypoxic signal. Furthermore, a novel double plasmid system was designed to elevate the potency of the vigilant vector. Instead of putting the promoter and the reporter gene in the same plasmid (single plasmid system), we separated them into two plasmids: the transactivator plasmid and reporter plasmid (double plasmid system). The hypoxia response element (HRE) incorporated promoter increased the expression of a chimeric transcription factor consisting of the yeast GAL4 DNA binding domain and the human nuclear (transcription) factor-kappaB (NF-kappaB) p65 activation domain. The powerful chimeric regulator binds specifically to the upstream activating sequence for GAL4 in the reporter plasmid and activates the transcription of the transgene. Our experiments showed that the HRE-mediated expression could quickly increase 2.08 +/- 0.75-fold within 6 hours of hypoxia and further augmented 7.12 +/- 1.52 fold when the hypoxia condition was prolonged to 24 hours. The hypoxia-inducible double plasmid system dramatically amplified the transgene expression under both hypoxia and normoxia by 412.79 +/- 185.27-fold and 205.35 +/- 65.44-fold, respectively, relative to the single plasmid system. From these results, we concluded that this hypoxia inducible double plasmid system could be used therapeutically to switch on genes that have proven beneficial effects in myocardial ischemia. PMID- 11882634 TI - Renal resistance index and progression of renal disease. AB - The progression of renal disease depends on various clinical parameters such as hypertension and proteinuria. We recently showed that an increased renal resistance index measured by duplex ultrasound is associated with a poor prognosis in patients with renal artery stenosis. We now prospectively tested the hypothesis that a high renal resistance index (greater-than-or-equal 80) predicts progression of renal disease in patients without renal artery stenosis. In 162 patients newly diagnosed with renal disease, the resistance index (1-[end diastolic velocity/maximum systolic velocity]*100) was measured in segmental arteries of both kidneys. Creatinine clearance was measured at baseline, at 3, 6, and 12 months, and then at yearly intervals thereafter (mean follow-up 3 +/- 1.4 years). The combined endpoint was a decrease of creatinine clearance by greater than-or-equal 50%, end-stage renal disease with replacement therapy, or death. Twenty-five patients (15%) had a renal resistance index value greater-than-or equal 80 at baseline. Nineteen (76%) had a decline in renal function; 16 (64%) progressed to dialysis, and 6 (24%) died. In comparison, in patients with renal resistance index values <80, 13 (9%) had a decline in renal function, only 7 (5%) became dialysis-dependent, and 2 (1%) died (P<0.001). In a multivariate regression analysis, only proteinuria and resistance index were independent predictors of declining renal function. A renal resistance index value of greater than-or-equal to 80 reliably identifies patients at risk for progressive renal disease. PMID- 11882635 TI - Functional significance of activation of calcium/calmodulin-dependent protein kinase II in angiotensin II--induced vascular hyperplasia and hypertension. AB - We have reported that norepinephrine (NE) and angiotensin II (Ang II) increase CaM kinase II activity, which, in turn, activates cytosolic phospholipase A(2) (PLA(2)) and releases arachidonic acid. The products of arachidonic acid generated via cytochrome P-450 and lipoxygenase contribute to the development of hypertension and vascular smooth muscle cell (VSMC) hyperplasia. The purpose of this study was to investigate whether CaM kinase II contributes to VSMC proliferation elicited by NE and Ang II and to hypertension induced by Ang II. NE (1 micromol/L) and Ang II (1 micromol/L) increased proliferation of rabbit aortic VSMC as measured by increased [(3)H]-thymidine incorporation; this effect of NE and Ang II was attenuated 88 +/- 10% and 64 +/- 11% by the CaM kinase II inhibitor KN-93, respectively. Infusion of Ang II with miniosmotic pumps (350 ng/min for 6 days) in rats elevated mean arterial pressure (MABP), which was reduced by simultaneous infusion of KN-93 (578 ng/min, for 6 days) (Ang II alone: MABP =174 +/- 3 mm Hg, n=12 versus Ang II + KN-93: MABP 123 +/- 5 mm Hg, n=4, P<0.05). Administration of KN-93 as a single bolus injection (16 mg/Kg), but not its vehicle, in Ang II--infused hypertensive animals also decreased MABP from 179 +/- 9 mm Hg to 109 +/- 6 mm Hg (n=5, P<0.05). CaM kinase II activity was increased in the kidney of Ang II--infused hypertensive animals compared with normotensive controls. Treatment with KN-93 reduced CaM kinase II activity and ameliorated the intravascular injury in the kidneys of Ang II--infused hypertensive rats. Our data indicate that CaM kinase activation represents an important component of the mechanism(s) initiating VSMC proliferation and the development and maintenance of Ang II--induced hypertension in rat. PMID- 11882636 TI - Blood pressure variability and organ damage in a general population: results from the PAMELA study (Pressioni Arteriose Monitorate E Loro Associazioni). AB - In hypertensive patients, 24-hour blood pressure (BP) variability (V) shows a positive relationship with organ damage, organ damage progression, and cardiovascular morbidity. The clinical relevance of BPV in the population has never been investigated. In a sample of 3200 individuals, randomly selected from the general population of Monza (Milan, Italy), we evaluated BP by an automatic oscillometric device every 20 minutes for 24 hours and left ventricular mass index (LVMI) by echocardiography. In each subject, individual systolic and diastolic BP readings were averaged to obtain a 24-hour mean. Systolic BPV was obtained by calculating (1) the standard deviation of the 24-hour mean, which was taken as the overall BPV, (2) the cyclic components (Fourier spectral analysis) that in the population as a whole explained >95% of the overall BPV, and (3) the fraction of the overall BPV that in each subject was not accounted for by the 2 cyclic components, termed individual residual BPV. A similar procedure was used for diastolic BP and heart rate. Participation rate was 64.1%. Patients receiving antihypertensive therapy (n=403) were excluded from the analysis, which was therefore limited to 1648 participants. In the population as a whole, LVMI significantly related to 24-hour systolic and diastolic BP mean (beta=0.40 and beta=0.37, respectively, P<0.001 for both) but not to the 2 cyclic components that accounted for most of the BPV. On the other hand, the individual residual BPV (which accounts on average for about 50% of overall BPV) showed a significant positive relationship with LVMI (beta =0.38 and beta=0.88 for systolic and diastolic BP, respectively, P<0.05 and P<0.01). No relationship was found between LVMI and heart rate values. These findings provide evidence that there is a relationship between LVMI and 24-hour average BP values in the population. They also provide the first demonstration that in the population there is also a positive independent association between LVMI and BPV. This association, however, can be exclusively seen with the BPV component that has an erratic rather than a cyclic nature. PMID- 11882637 TI - Renal hemodynamic control by endothelin and nitric oxide under angiotensin II blockade in man. AB - To investigate whether endothelin-A receptors and nitric oxide modulate renal hemodynamics in man under angiotensin II receptor-1 blockade, 6 healthy volunteers, on a 240 mmol Na diet, underwent 4 separate renal hemodynamic measurements, in 3 of which endothelin-A blocker BQ-123 0.2 nmol.kg.min(-1) was infused for 90 minutes after pretreatment with either placebo, telmisartan 1 mg.kg center dot day(-1) for 3 days, or telmisartan as well, but with co-infusion of both BQ-123 and N(G)-nitro-L-arginine methylester 0.5 microg.kg center dot min(-1). A fourth infusion was made with N(G)-nitro-L-arginine methylester alone. No change followed infusion of either N(G)-nitro-L-arginine methylester alone or BQ-123 alone. With BQ-123 after telmisartan, renal blood flow rose from 916 +/- 56 mL center dot min(-1) center dot 1.73 m(2) to 1047 +/- 51.2 (P<0.001), and renal vascular resistances fell from 89 +/- 7 mm Hg center dot min center dot L( 1) to 74 +/-4 (P<0.001). These changes were fully abolished by the co-infused N(G)-nitro-L-arginine methylester. Infusion of BQ-123, devoid of renal hemodynamic effects at baseline, produces significant renal vasodilation when angiotensin II receptors are blocked, indicating an increasing renal hemodynamic role of endothelin-A--receptor activity. Because such a vasodilation is prevented by nonvasoconstricting microdoses of N(G)-nitro-L-arginine methylester, nitric oxide--endothelin balance controls substantially renal hemodynamics under angiotensin II blockade. These findings are consistent with a rationale of the association of endothelin-A blockers with angiotensin II blockers or angiotensin converting enzyme inhibitors in treating nitric oxide--deficient conditions such as arterial hypertension, heart failure, and chronic renal diseases. PMID- 11882645 TI - Enzymatic modification of tRNAs: MiaB is an iron-sulfur protein. AB - The product of the miaB gene, MiaB, from Escherichia coli participates in the methylthiolation of the adenosine 37 residue during modification of tRNAs that read codons beginning with uridine. A His-tagged version of MiaB has been overproduced and purified to homogeneity. Gel electrophoresis and size exclusion chromatography revealed that MiaB protein is a monomer. As isolated MiaB contains both iron and sulfide and an apoprotein form can chelate as much as 2.5-3 iron and 3-3.5 sulfur atoms per polypeptide chain. UV-visible and EPR spectroscopy of MiaB indicate the presence of a [4Fe-4S] cluster under reducing and anaerobic conditions, whereas [2Fe-2S] and [3Fe-4S] forms are generated under aerobic conditions. Preliminary site-directed mutagenesis studies suggest that Cys(157), Cys(161), and Cys(164) are involved in iron chelation and that the cluster is essential for activity. Together with the previously shown requirement of S adenosylmethionine (AdoMet) for the methylthiolation reaction, the finding that MiaB is an iron-sulfur protein suggests that it belongs to a superfamily of enzymes that uses [Fe-S] centers and AdoMet to initiate radical catalysis. MiaB is the first and only tRNA modification enzyme known to contain an Fe-S cluster. PMID- 11882646 TI - Identification of a neuronal Cdk5 activator-binding protein as Cdk5 inhibitor. AB - Neuronal Cdc2-like kinase (Nclk) plays an important role in a variety of cellular processes, including neuronal cell differentiation, apoptosis, neuron migration, and formation of neuromuscular junction. The active kinase consists of a catalytic subunit, Cdk5, and an essential regulatory subunit, neuronal Cdk5 activator (p35(nck5a) or p25(nck5a)), which is expressed primarily in neurons of central nervous tissue. In our previous study using the yeast two-hybrid screening method, three novel p35(nck5a)-associated proteins were isolated. Here we show that one of these proteins, called C42, specifically inhibits the activation of Cdk5 by Nck5a. Co-immunoprecipitation data suggested that C42 and p35(nck5a) could form a complex within cultured mammalian cells. Deletion analysis has mapped the inhibitory domain of C42 to a region of 135 amino acids, which is conserved in Pho81, a yeast protein that inhibits the yeast cyclin dependent protein kinase Pho85. The Pho85.Pho80 kinase complex has been shown to be the yeast functional homologue of the mammalian Cdk5/p35(nck5a) kinase. PMID- 11882647 TI - Transcript abundance in yeast varies over six orders of magnitude. AB - In the current era of functional genomics, it is remarkable that the intracellular range of transcript abundance is largely unknown. For the yeast Saccharomyces cerevisiae, hybridization-based complexity analysis and SAGE analysis showed that the majority of yeast mRNAs are present at one or fewer copies per cell; however, neither method provides an accurate estimate of the full range of low abundance transcripts. Here we examine the range of intracellular transcript abundance in yeast using kinetically monitored, reverse transcriptase-initiated PCR (kRT-PCR). Steady-state transcript levels encoded by all 65 genes on the left arm of chromosome III and 185 transcription factor genes are quantitated. Abundant transcripts encoded by glycolytic genes, previously quantitated by kRT-PCR, are present at a few hundred copies per cell whereas genes encoding physiologically important transcription factors are expressed at levels as low as one-thousandth transcript per cell. Of the genes assessed, only the silent mating type loci, HML and HMR, are transcriptionally silent. The results show that transcript abundance in yeast varies over six orders of magnitude. Finally, kRT-PCR, cDNA microarray, and high density oligonucleotide array assays are compared for their ability to detect and quantitate the complete yeast transcriptome. PMID- 11882648 TI - Interaction of the Salmonella typhimurium transcription and virulence factor SlyA with target DNA and identification of members of the SlyA regulon. AB - The SlyA protein from Salmonella typhimurium is a transcription factor that contributes to virulence. It is shown that a slyA mutant is attenuated in the presence of murine macrophages compared with the parent strain. Moreover, after growth in minimal medium, survival of the slyA mutant was reduced. Altered levels of flagellin (fliC), PagC, IroN, and outer membrane proteins suggest that the slyA mutation affects the surface properties of Salmonella. The isolated SlyA protein is a cofactor-free homodimer that recognizes five sites within the promoter region of the slyA gene. One of these sites contained a near perfect inverted repeat TTAGCAAGCTAA. The other four sites contained related sequences. Occupation of the SlyA sites in the slyA promoter prevented open-complex formation, consistent with the pattern of slyA::lacZ expression parental and slyA mutant strains. By combining the footprinting data with potential SlyA binding sites recovered from a pool of random DNA sequences, a consensus was defined and used to probe the NIH Salmonella unfinished genomes data base. These searches revealed the presence of consensus SlyA sites upstream of omp, ispA, xseB, slyA, and a gene encoding a protein with homology to a hemagglutinin. Accordingly, transcription of an omp::lacZ fusion was reduced in a slyA mutant. Given the difficulties in obtaining a comprehensive picture of intracellular gene expression, the definition of the DNA sequence recognized by a transcription factor (SlyA) that is essential for survival in the macrophage environment should allow a complete regulon of genes with altered expression upon exposure to macrophages to be determined once the S. typhimurium genome annotation is complete. PMID- 11882649 TI - Identification of the heparan sulfate binding sites in the cellular prion protein. AB - Data from cell culture and animal models of prion disease support the separate involvement of both heparan sulfate proteoglycans and copper (II) ions in prion (PrP) metabolism. Though direct interactions between prion protein and heparin have been recorded, little is known of the structural features implicit in this interaction or of the involvement of copper (II) ions. Using biosensor and enzyme linked immunosorbent assay methodology we report direct heparin and heparan sulfate-binding activity in recombinant cellular prion protein (PrP(c)). We also demonstrate that the interaction of recombinant PrP(c) with heparin is weakened in the presence of Cu(II) ions and is particularly sensitive to competition with dextran sulfate. Competitive inhibition experiments with chemically modified heparins also indicate that 2-O-sulfate groups (but not 6-O-sulfate groups) are essential for heparin recognition. We have also identified three regions of the prion protein capable of independent binding to heparin and heparan sulfate: residues 23-52, 53-93, and 110-128. Interestingly, the interaction of an octapeptide-spanning peptide motif amino acids 53-93 with heparin is enhanced by Cu(II) ions. Significantly, a peptide of this sequence is able to inhibit the binding of full-length prion molecule to heparin, suggesting a direct role in heparin recognition within the intact protein. The collective data suggest a complex interaction between prion protein and heparin/heparan sulfate and has implications for the cellular and pathological functions of prion proteins. PMID- 11882650 TI - Molecular characterization of mammalian dicarbonyl/L-xylulose reductase and its localization in kidney. AB - In this report, we first cloned a cDNA for a protein that is highly expressed in mouse kidney and then isolated its counterparts in human, rat hamster, and guinea pig by polymerase chain reaction-based cloning. The cDNAs of the five species encoded polypeptides of 244 amino acids, which shared more than 85% identity with each other and showed high identity with a human sperm 34-kDa protein, P34H, as well as a murine lung-specific carbonyl reductase of the short-chain dehydrogenase/reductase superfamily. In particular, the human protein is identical to P34H, except for one amino acid substitution. The purified recombinant proteins of the five species were about 100-kDa homotetramers with NADPH-linked reductase activity for alpha-dicarbonyl compounds, catalyzed the oxidoreduction between xylitol and l-xylulose, and were inhibited competitively by n-butyric acid. Therefore, the proteins are designated as dicarbonyl/l xylulose reductases (DCXRs). The substrate specificity and kinetic constants of DCXRs for dicarbonyl compounds and sugars are similar to those of mammalian diacetyl reductase and l-xylulose reductase, respectively, and the identity of the DCXRs with these two enzymes was demonstrated by their co-purification from hamster and guinea pig livers and by protein sequencing of the hepatic enzymes. Both DCXR and its mRNA are highly expressed in kidney and liver of human and rodent tissues, and the protein was localized primarily to the inner membranes of the proximal renal tubules in murine kidneys. The results imply that P34H and diacetyl reductase (EC ) are identical to l-xylulose reductase (EC ), which is involved in the uronate cycle of glucose metabolism, and the unique localization of the enzyme in kidney suggests that it has a role other than in general carbohydrate metabolism. PMID- 11882651 TI - Liver glycogen synthase but not the muscle isoform differentiates between glucose 6-phosphate produced by glucokinase or hexokinase. AB - Using adenovirus-mediated gene transfer into FTO-2B cells, a rat hepatoma cell line, we have overexpressed hexokinase I (HK I), glucokinase (GK), liver glycogen synthase (LGS), muscle glycogen synthase (MGS), and combinations of each of the two glucose-phosphorylating enzymes with each one of the GS isoforms. FTO-2B cells do not synthesize glycogen even when incubated with high doses of glucose. Adenovirus-induced overexpression of HK I and/or LGS, two enzymes endogenously expressed by these cells, did not produce a significant increase in the levels of active GS and the total glycogen content. In contrast, GK overexpression led to the glucose-dependent activation of endogenous or overexpressed LGS and to the accumulation of glycogen. Similarly overexpressed MGS was efficiently activated by the glucose-6-phosphate (Glc-6-P) produced by either endogenous or overexpressed HK I and by overexpressed GK. These results indicate the existence of at least two pools of Glc-6-P in the cell, one of them is accessible to both isoforms of GS and is replenished by the action of GK, whereas LGS is excluded from the cellular compartment where the Glc-6-P produced by HK I is directed. These findings are interpreted in terms of the metabolic role that the two pairs of enzymes, HK I-MGS in the muscle and GK-LGS in the hepatocyte, perform in their respective tissues. PMID- 11882652 TI - Signaling events in amyloid beta-peptide-induced neuronal death and insulin-like growth factor I protection. AB - Amyloid beta-peptide (Abeta) is implicated as the toxic agent in Alzheimer's disease and is the major component of brain amyloid plaques. In vitro, Abeta causes cell death, but the molecular mechanisms are unclear. We analyzed the early signaling mechanisms involved in Abeta toxicity using the SH-SY5Y neuroblastoma cell line. Abeta caused cell death and induced a 2- to 3-fold activation of JNK. JNK activation and cell death were inhibited by overexpression of a dominant-negative SEK1 (SEK1-AL) construct. Butyrolactone I, a cdk5 inhibitor, had an additional protective effect against Abeta toxicity in these SEK1-AL-expressing cells suggesting that cdk5 and JNK activation independently contributed to this toxicity. Abeta also weakly activated ERK and Akt but had no effect on p38 kinase. Inhibitors of ERK and phosphoinositide 3-kinase (PI3K) pathways did not affect Abeta-induced cell death, suggesting that these pathways were not important in Abeta toxicity. Insulin-like growth factor I protected against Abeta toxicity by strongly activating ERK and Akt and blocking JNK activation in a PI3K-dependent manner. Pertussis toxin also blocked Abeta-induced cell death and JNK activation suggesting that G(i/o) proteins were upstream activators of JNK. The results suggest that activation of the JNK pathway and cdk5 may be initial signaling cascades in Abeta-induced cell death. PMID- 11882653 TI - The effects of mutations in helices 4 and 6 of ApoA-I on scavenger receptor class B type I (SR-BI)-mediated cholesterol efflux suggest that formation of a productive complex between reconstituted high density lipoprotein and SR-BI is required for efficient lipid transport. AB - We have studied the effects of mutations in apoA-I on reconstituted high density lipoprotein (HDL) particle (rHDL(apoA-I)) binding to and cholesterol efflux from wild-type (WT) and mutant forms of the HDL receptor SR-BI expressed by ldlA-7 cells. Mutations in helix 4 or helix 6 of the apoA-I reduced efflux by 79 and 51%, respectively, without substantially altering receptor binding (apparent K(d) values of 1.1-4.4 microg of protein/ml). SR-BI with an M158R mutation bound poorly to rHDL with WT and helix 4 mutant apoA-I; the helix 6 mutant restored tight binding to SR-BI(M158R) (K(d) values of 48, 60, and 7 microg of protein/ml, respectively). SR-BI(M158R)-mediated cholesterol efflux rates, normalized for binding, were high for all three rHDLs (71-111% of control). In contrast, absolute (12-19%) and binding-corrected (24-47%) efflux rates for all three rHDLs mediated by SR-BI with Q402R/Q418R mutations were very low. We propose that formation of a productive complex between apoA-I in rHDL and SR-BI, in which the lipoprotein and the receptor must either be precisely aligned or have the capacity to undergo appropriate conformational changes, is required for efficient SR-BI-mediated cholesterol efflux. Some mutations in apoA-I and/or SR-BI can result in high affinity, but non-productive, binding that does not permit efficient cholesterol efflux. PMID- 11882654 TI - Identification of a karyopherin alpha 2 recognition site in PLAG1, which functions as a nuclear localization signal. AB - The activation of the pleomorphic adenoma gene 1 (PLAG1) is the most frequent gain-of-function mutation found in pleomorphic adenomas of the salivary glands. To gain more insight into the regulation of PLAG1 function, we searched for PLAG1 interacting proteins. Using the yeast two-hybrid system, we identified karyopherin alpha2 as a PLAG1-interacting protein. Physical interaction between PLAG1 and karyopherin alpha2 was confirmed by an in vitro glutathione S transferase pull-down assay. Karyopherin alpha2 escorts proteins into the nucleus via interaction with a nuclear localization sequence (NLS) composed of short stretches of basic amino acids. Two putative NLSs were identified in PLAG1. The predicted NLS1 (KRKR) was essential for physical interaction with karyopherin alpha2 in glutathione S-transferase pull-down assay, and its mutation resulted in decreased nuclear import of PLAG1. Moreover, NLS1 was able to drive the nuclear import of the cytoplasmic protein beta-galactosidase. In contrast, predicted NLS2 of PLAG1 (KPRK) was not involved in karyopherin alpha2 binding nor in its nuclear import. The residual nuclear import of PLAG1 after mutation of the NLS1 was assigned to the zinc finger domain of PLAG1. These observations indicate that the nuclear import of PLAG1 is governed by its zinc finger domain and by NLS1, a karyopherin alpha2 recognition site. PMID- 11882655 TI - Recombinant class I aldehyde dehydrogenases specific for all-trans- or 9-cis retinal. AB - The molecular basis for the specificity of aldehyde dehydrogenases (ALDHs) for retinal, the precursor of the morphogen retinoic acid, is still poorly understood. We have expressed in Escherichia coli both retinal dehydrogenase (RALDH), a cytosolic aldehyde dehydrogenase originally isolated from rat kidney, and the highly homologous phenobarbital-induced aldehyde dehydrogenase (PB-ALDH). Oxidation of propanal was observed with both enzymes. On the other hand, recombinant RALDH efficiently catalyzed oxidation of 9-cis- and all-trans retinal, whereas PB-ALDH was inactive with all-trans-retinal and poorly active with 9-cis-retinal. A striking difference between PB-ALDH and all other class I ALDHs is the identity of the amino acid immediately preceding the active nucleophile Cys(302) (Ile(301) instead of Cys(301)). Nevertheless, these amino acids could be exchanged in either RALDH or PB-ALDH without affecting substrate specificity. Characterization of chimeric enzymes demonstrates that distinct groups of amino acids control the differential activity of RALDH and PB-ALDH with all-trans- and 9-cis-retinal. Of 52 divergent amino acids, the first 17 are crucial for activity with all-trans-retinal, whereas the next 25 are important for catalysis of 9-cis-retinal oxidation. Recombinant enzymes with specificity for all-trans- or 9-cis-retinal were obtained, which should provide useful tools to study the relative importance of local production of all-trans- versus 9-cis retinoic acid in development and tissue differentiation. PMID- 11882656 TI - Regulation of endocytosis of activin type II receptors by a novel PDZ protein through Ral/Ral-binding protein 1-dependent pathway. AB - Using yeast two-hybrid screening, we have identified a mouse Postsynaptic density 95/Discs large/Zona occludens-1 (PDZ) protein that interacts with activin type II receptors (ActRIIs). We named the protein activin receptor-interacting protein 2 (ARIP2). ARIP2 was found to have one PDZ domain in the NH(2)-terminal region and interact specifically with ActRIIs among the receptors for the transforming growth factor beta family by the PDZ domain. Interestingly, overexpression of ARIP2 enhances endocytosis of ActRIIs and reduces activin-induced transcription in Chinese hamster ovary K1 cells. In addition, immunofluorescence co localization studies indicated the direct involvement of ARIP2 in the intracellular translocation of ActRIIs by PDZ domain-mediated interaction. Moreover, we have identified that the COOH-terminal region of ARIP2 interacts with Ral-binding protein 1 (RalBP1). RalBP1 is a potential effector protein of small GTP-binding protein Ral and regulates endocytosis of epidermal growth factor and insulin receptors. The studies using deletion mutants of RalBP1 and constitutively GTP and GDP binding forms of Ral indicate that ARIP2 regulates endocytosis of ActRIIs through the Ral/RalBP1-dependent pathway, and the GDP-GTP exchange of Ral is critical for this regulation. PMID- 11882657 TI - Functional classification of ADAMs based on a conserved motif for binding to integrin alpha 9beta 1: implications for sperm-egg binding and other cell interactions. AB - ADAMs (a disintegrin and metalloproteases) are members of the metzincin superfamily of metalloproteases. Among integrins binding to disintegrin domains of ADAMs are alpha(9)beta(1) and alpha(v)beta(3), and they bind in an RGD independent and an RGD-dependent manner, respectively. Human ADAM15 is the only ADAM with the RGD motif in the disintegrin domain. Thus, both integrin alpha(9)beta(1) and alpha(v)beta(3) recognize the ADAM15 disintegrin domain. We determined how these integrins recognize the ADAM15 disintegrin domain by mutational analysis. We found that the Arg(481) and the Asp-Leu-Pro-Glu-Phe residues (residues 488-492) were critical for alpha(9)beta(1) binding, but the RGD motif (residues 484-486) was not. In contrast, the RGD motif was critical for alpha(v)beta(3) binding, but the other residues flanking the RGD motif were not. As the RX(6)DLPEF alpha(9)beta(1) recognition motif (residues 481-492) is conserved among ADAMs, except for ADAM10 and 17, we hypothesized that alpha(9)beta(1) may recognize disintegrin domains in all ADAMs except ADAM10 and 17. Indeed we found that alpha(9)beta(1) bound avidly to the disintegrin domains of ADAM1, 2, 3, and 9 but not to the disintegrin domains of ADAM10 and 17. As several ADAMs have been implicated in sperm-oocyte interaction, we tested whether the functional classification of ADAMs, based on specificity for integrin alpha(9)beta(1), applies to sperm-egg binding. We found that the ADAM2 and 15 disintegrin domains bound to oocytes, but the ADAM17 disintegrin domain did not. Furthermore, the ADAM2 and 15 disintegrin domains effectively blocked binding of sperm to oocytes, but the ADAM17 disintegrin domain did not. These results suggest that oocytes and alpha(9)beta(1) have similar binding specificities for ADAMs and that alpha(9)beta(1), or a receptor with similar specificity, may be involved in sperm-egg interaction during fertilization. As alpha(9)beta(1) is a receptor for many ADAM disintegrins and alpha(9)beta(1) and ADAMs are widely expressed, alpha(9)beta(1)-ADAM interaction may be of a broad biological importance. PMID- 11882658 TI - Molecular dissection of the functional domains of a unique, tartrate-resistant, surface membrane acid phosphatase in the primitive human pathogen Leishmania donovani. AB - The primitive trypanosomatid pathogen of humans, Leishmania donovani, constitutively expresses a unique externally oriented, tartrate-resistant, acid phosphatase on its surface membrane. This is of interest because these organisms are obligate intracellular protozoan parasites that reside and multiply within the hydrolytic milieu of mammalian macrophage phago-lysosomes. Here we report the identification of the gene encoding this novel L. donovani enzyme. In addition, we characterized its structure, demonstrated its constitutive expression in both parasite developmental forms, and determined the cell surface membrane localization of its translated protein product. Further, we used a variety of green fluorescent protein chimeric constructs as reporters in a homologous leishmanial expression system to dissect the functional domains of this unique, tartrate-resistant, surface membrane enzyme. PMID- 11882659 TI - A combination of cis-acting elements is required to activate the pro-alpha 1(I) collagen promoter in tendon fibroblasts of transgenic mice. AB - The genes encoding the two type I collagen chains are selectively activated in few cell types, including fibroblasts and osteoblasts. By generating transgenic mice, we have previously shown that the activity of the mouse pro-alpha1(I) promoter was controlled by separate cell-specific cis-acting elements. In particular, a sequence located between -3.2 and -2.3 kb was needed to induce expression of the reporter gene at high levels in tendon fibroblasts. In the present work, by using the same transgenic approach, we have identified two short elements in this sequence, named tendon-specific element (TSE) 1 and TSE2, that were necessary to direct reporter gene expression selectively in tendon fibroblasts. Gel shift assays showed that TSE1 and TSE2 bound proteins specifically present in nuclear extracts from tendon fibroblasts and that the sequence of TSE2 binding a tendon-specific protein corresponded to an E-box. Analysis of transgenic mice further indicated that TSE1 and TSE2 needed to cooperate not only with each other but also with other cis-acting elements of the proximal promoter to activate reporter gene expression in tendon fibroblasts. Similarly, it pointed out that the so-called osteoblast-specific element had to interact with downstream sequences to drive reporter gene expression in osteoblasts of transgenic mice. Thus, expression of the mouse pro-alpha1(I) collagen gene in tendon fibroblasts appears to be the result of a unique combination of different cis-acting elements, including TSE1 and TSE2. PMID- 11882660 TI - Regulation of protein S-thiolation by glutaredoxin 5 in the yeast Saccharomyces cerevisiae. AB - The irreversible oxidation of cysteine residues can be prevented by protein S thiolation, a process by which protein -SH groups form mixed disulfides with low molecular weight thiols such as glutathione. We report here that this protein modification is not a simple response to the cellular redox state, since different oxidants lead to different patterns of protein S-thiolation. SDS polyacrylamide gel electrophoresis shows that glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is the major target for modification following treatment with hydroperoxides (hydrogen peroxide or tert-butylhydroperoxide), whereas this enzyme is unaffected following cellular exposure to the thiol oxidant diamide. Further evidence that protein S-thiolation is tightly regulated in response to oxidative stress is provided by the finding that the Tdh3 GAPDH isoenzyme, and not the Tdh2 isoenzyme, is S-thiolated following exposure to H(2)O(2) in vivo, whereas both GAPDH isoenzymes are S-thiolated when H(2)O(2) is added to cell-free extracts. This indicates that cellular factors are likely to be responsible for the difference in GAPDH S-thiolation observed in vivo rather than intrinsic structural differences between the GAPDH isoenzymes. To begin to search for factors that can regulate the S-thiolation process, we investigated the role of the glutaredoxin family of oxidoreductases. We provide the first evidence that protein dethiolation in vivo is regulated by a monothiol-glutaredoxin rather than the classical glutaredoxins, which contain two active site cysteine residues. In particular, glutaredoxin 5 is required for efficient dethiolation of the Tdh3 GAPDH isoenzyme. PMID- 11882661 TI - Tandem UAA repeats at the 3'-end of the transcript are essential for the precise initiation of reverse transcription of the I factor in Drosophila melanogaster. AB - Non-long terminal repeat retrotransposons, widespread among eukaryotic genomes, transpose by reverse transcription of an RNA intermediate. Some of them, like L1 in the human, terminate at the 3'-end with a poly(dA) stretch whereas others, like the I factor in Drosophila melanogaster, have instead a short sequence repeated in tandem. This suggests different requirements for the initiation of reverse transcription. Here, we have used an RNA circularization/reverse transcription-PCR technique to analyze the 5'- and 3'-ends of the full-length transcripts produced by the I factor at the time of active retrotransposition. These transcripts are capped and polyadenylated similar to conventional messenger RNAs. We have analyzed the 3'-ends of transcripts and transposed copies produced by I elements mutated at the 3'-ends. Transcripts devoid of tandem UAA repeats, although capable of building the components of the retrotransposition machinery, are inefficiently used as retrotransposition intermediates. Such transcripts produce rare new integrated copies issued from the inaccurate initiation of reverse transcription near the 3'-end of the element. The tandem UAA repeats at the 3'-end of the transcripts of I are required for the efficient and precise initiation of reverse transcription. This strong specificity of the I factor reverse transcriptase for its own transcript has implications for the impact of I factor retrotransposition on the host genome. PMID- 11882662 TI - The interaction of RGSZ1 with SCG10 attenuates the ability of SCG10 to promote microtubule disassembly. AB - RGS proteins (regulators of G protein signaling) are a diverse family of proteins that act to negatively regulate signaling by heterotrimeric G proteins. Initially characterized as GTPase-activating proteins for Galpha subunits, recent data have implied additional functions for RGS proteins. We previously identified an RGS protein (termed RGSZ1) whose expression is quite specific to neuronal tissue (Glick, J. L., Meigs, T. E., Miron, A., and Casey, P. J. (1998) J. Biol. Chem. 273, 26008-26013). In a continuing effort to understand the role of RGSZ1 in cellular signaling, the yeast two-hybrid system was employed to identify potential effector proteins of RGSZ1. The microtubule-destabilizing protein SCG10 (superior cervical ganglia, neural specific 10) was found to directly interact with RGSZ1 in the yeast system, and this interaction was further verified using direct binding assays. Treatment of PC12 cells with nerve growth factor resulted in Golgi-specific distribution of SCG10. A green fluorescent protein-tagged variant of RGSZ1 translocated to the Golgi complex upon treatment of PC12 cells with nerve growth factor, providing evidence that RGSZ1 and SCG10 interact in cells as well as in vitro. Analysis of in vitro microtubule polymerization/depolymerization showed that binding of RGSZ1 to SCG10 effectively blocked the ability of SCG10 to induce microtubule disassembly as determined by both turbidimetric and microscopy-based assays. These results identify a novel connection between RGS proteins and the cytoskeletal network that points to a broader role than previously envisioned for RGS proteins in regulating biological processes. PMID- 11882663 TI - Structural determinants of the Na+/H+ exchanger regulatory factor interaction with the beta 2 adrenergic and platelet-derived growth factor receptors. AB - The Na(+)/H(+) exchanger regulatory factor (NHERF) binds through its PDZ1 domain to the carboxyl-terminal sequences NDSLL and EDSFL of the beta(2) adrenergic receptor (beta(2)AR) and platelet-derived growth factor receptor, respectively, and plays a critical role in the membrane localization and physiological regulation of these receptors. The crystal structures of the human NHERF PDZ1 domain bound to the sequences NDSLL and EDSFL have been determined at 1.9- and 2.2-A resolution, respectively. The beta(2)AR and platelet-derived growth factor receptor ligands insert into the PDZ1 binding pocket by a beta-sheet augmentation process and are stabilized by largely similar networks of hydrogen bonds and hydrophobic contacts. In the PDZ1-beta(2)AR complex, the side chain of asparagine at position -4 in the beta(2)AR peptide forms two additional hydrogen bonds with Gly(30) of PDZ1, which contribute to the higher affinity of this interaction. Remarkably, both complexes are further stabilized by hydrophobic interactions involving the side chains of the penultimate amino acids of the peptide ligands, whereas the PDZ1 residues Asn(22) and Glu(43) undergo conformational changes to accommodate these side chains. These results provide structural insights into the mechanisms by which different side chains at the position -1 of peptide ligands interact with PDZ domains and contribute to the affinity of the PDZ-ligand interaction. PMID- 11882664 TI - Specific recognition and cleavage of galectin-3 by Leishmania major through species-specific polygalactose epitope. AB - Lipophosphoglycan is a major surface molecule of Leishmania, protozoa parasites, which are the causative agents of leishmaniasis, a disease that annually afflicts millions of people worldwide. The oligosaccharide structures of lipophosphoglycan varies among species, and epitopes of these species-specific oligosaccharides are suggested to be implicated in the interaction of Leishmania with macrophages as well as species-specific tissue tropism observed in leishmaniasis. The recognition of the species-specific variation of oligosaccharides is likely to be mediated by host carbohydrate-binding proteins, lectins, but the identities of the lectins remain elusive. Galectin-3 is a mammalian soluble beta-galactoside binding lectin and is expressed in macrophages, dendritic cells, and keratinocytes, as well as fibroblasts, all of which are present in the site of Leishmania infection. In this paper, we found that galectin-3 binds to lipophosphoglycan of Leishmania major but not to those of Leishmania donovani through L. major-specific polygalactose epitopes. Association of galectin-3 with L. major led to the cleavage of galectin-3, resulting in truncated galectin-3 containing the C-terminal lectin domain but lacking the N-terminal domain implicated in lectin oligomerization. This cleavage was inhibited by the galectin 3 antagonist lactose, as well as 1,10-ortho-phenanthroline, suggesting that galectin-3 is cleaved by zinc metalloproteases after its binding to lipophosphoglycans. The modulation of various innate immunity reactions by galectin-3 is affected by its oligomerization; therefore, we propose the L. major specific truncation of galectin-3 may contribute to the species-specific immune responses induced by Leishmania. PMID- 11882665 TI - Zinc cluster protein Rdr1p is a transcriptional repressor of the PDR5 gene encoding a multidrug transporter. AB - The yeast PDR5 gene encodes an efflux pump that confers multidrug resistance. Expression of PDR5 is positively regulated by the transcription factors Pdr1p and Pdr3p that recognize the same pleiotropic drug resistance elements (PDREs) in the PDR5 promoter. Pdr1p and Pdr3p belong to the Gal4p family of zinc cluster proteins. The function of RDR1 (YOR380W), which also encodes a member of this family, is unknown. To identify target genes for Rdr1p, we have performed whole genome analysis of gene expression with DNA microarrays. Our results show that Rdr1p is a transcriptional repressor of five genes, including PDR5. A Deltardr1 strain has increased resistance to cycloheximide, as expected from the overexpression of PDR5. In addition, the activity of a PDR5-lacZ reporter is increased in a Deltardr1 strain. All (but one) genes affected by removal of Rdr1p contain PDREs in their promoters. We tested if the effect of Rdr1p is mediated through PDREs by inserting this DNA element in front of a minimal promoter. Activity of this reporter was increased in a Deltardr1 strain. Moreover, mutations known to reduce binding of Pdr1/Pdr3p abolished the induction observed in the Deltardr1 strain. Thus, we have identified a transcriptional repressor involved in the control of multidrug resistance. PMID- 11882666 TI - The perplexing challenges of a pump turned channel. PMID- 11882667 TI - Synaptophysins: vesicular cation channels? PMID- 11882669 TI - The role of histidine residues in modulation of the rat P2X(2) purinoceptor by zinc and pH. AB - P2X(2) receptor currents are potentiated by acidic pH and zinc. To identify residues necessary for proton and zinc modulation, alanines were singly substituted for each of the nine histidines in the extracellular domain of the rat P2X(2) receptor. Wild-type and mutant receptors were expressed in Xenopus oocytes and analysed with two-electrode voltage clamp. All mutations caused less than a 2-fold change in the EC(50) of the ATP concentration-response relation. Decreasing the extracellular pH from 7.5 to 6.5 potentiated the responses to 10 microM ATP of wild-type P2X(2) and eight mutant receptors more than 4-fold, but the response of the mutant receptor H319A was potentiated only 1.4-fold. The H319A mutation greatly attenuated the maximal potentiation that could be produced by a drop in pH, shifted the pK(a) (-log of dissociation constant) of the potentiation to a more basic pH as compared with P2X(2) and revealed a substantial pH-dependent decrease in the maximum response with a pK(a) near 6.0. Substituting a lysine for H319 reduced the EC(50) for ATP 40-fold. Zinc (20 microM) potentiated the responses to 10 microM ATP of wild-type P2X(2) and seven histidine mutants by approximately 8-fold but had virtually no effect on the responses of two mutants, H120A and H213A. Neither H120A nor H213A removed the voltage-independent inhibition caused by high concentrations of zinc. The observation that different mutations selectively eliminated pH or zinc potentiation implies that there are two independent sites of action, even though the mechanisms of pH and zinc potentiation appear similar. PMID- 11882668 TI - Mutation of Walker-A lysine 464 in cystic fibrosis transmembrane conductance regulator reveals functional interaction between its nucleotide-binding domains. AB - The cystic fibrosis transmembrane conductance regulator (CFTR) chloride channel bears two nucleotide-binding domains (NBD1 and NBD2) that control its ATP dependent gating. Exactly how these NBDs control gating is controversial. To address this issue, we examined channels with a Walker-A lysine mutation in NBD1 (K464A) using the patch clamp technique. K464A mutants have an ATP dependence (EC(50) approximate 60 microM) and opening rate at 2.75 mM ATP (approximately 2.1 s(-1)) similar to wild type (EC(50) approximate 97 microM; approximately 2.0 s( 1)). However, K464A's closing rate at 2.75 mM ATP (approximately 3.6 s(-1)) is faster than that of wild type (approximately 2.1 s(-1)), suggesting involvement of NBD1 in nucleotide-dependent closing. Delay of closing in wild type by adenylyl imidodiphosphate (AMP-PNP), a non-hydrolysable ATP analogue, is markedly diminished in K464A mutants due to reduction in AMP-PNP's apparent on-rate and acceleration of its apparent off-rate (approximately 2- and approximately 10 fold, respectively). Since the delay of closing by AMP-PNP is thought to occur via NBD2, K464A's effect on the NBD2 mutant K1250A was examined. In sharp contrast to K464A, K1250A single mutants exhibit reduced opening (approximately 0.055 s(-1)) and closing (approximately 0.006 s(-1)) rates at millimolar [ATP], suggesting a role for K1250 in both opening and closing. At millimolar [ATP], K464A-K1250A double mutants close approximately 5-fold faster (approximately 0.029 s(-1)) than K1250A but open with a similar rate (approximately 0.059 s( 1)), indicating an effect of K464A on NBD2 function. In summary, our results reveal that both of CFTR's functionally asymmetric NBDs participate in nucleotide dependent closing, which provides important constraints for NBD-mediated gating models. PMID- 11882670 TI - Substrate-induced regulation of the human colonic monocarboxylate transporter, MCT1. AB - Butyrate is the principal source of energy for colonic epithelial cells, and has profound effects on their proliferation, differentiation and apoptosis. Transport of butyrate across the colonocyte luminal membrane is mediated by the monocarboxylate transporter 1 (MCT1). We have examined the regulation of expression of human colonic MCT1 by butyrate, in cultured colonic epithelial cells (AA/C1). Treatment with sodium butyrate (NaBut) resulted in a concentration and time-dependent upregulation of both MCT1 mRNA and protein. At 2 mM butyrate, the magnitude of induction of mRNA (5.7-fold) entirely accounted for the 5.2-fold increase in protein abundance, and was mediated by both activation of transcription and enhanced mRNA stability. The other monocarboxylates found naturally in the colon, acetate and propionate, had no effect. The properties of butyrate uptake by AA/C1 cells were characteristic of MCT1. Induction of the MCT1 protein resulted in a corresponding increase in the maximal rate of butyrate transport. The V(max) for uptake of [U-(14)C]butyrate was increased 5-fold following pre-incubation with 2 mM NaBut, with no significant change in the apparent K(m). In conclusion, this study is the first to show substrate-induced regulation of human colonic MCT1. The basis of this regulation is a butyrate induced increase in MCT1 mRNA abundance, resulting from the dual control of MCT1 gene transcription and stability of the MCT1 transcript. We suggest that butyrate induced increases in the expression and resulting activity of MCT1 serve as a mechanism to maximise intracellular availability of butyrate, to act both as a source of energy and to influence processes maintaining cellular homeostasis in the colonic epithelium. PMID- 11882671 TI - Functional characterization of recombinant human ClC-4 chloride channels in cultured mammalian cells. AB - Members of the ClC chloride channel family participate in several physiological processes and are linked to human genetic diseases. The physiological role of ClC 4 is unknown and previous detailed characterizations of recombinant human ClC-4 (hClC-4) have provided conflicting results. To re-examine the hClC-4 phenotype, recombinant hClC-4 was expressed in three distinct mammalian cell lines and characterized using patch-clamp techniques. In all cells, the expression of hClC 4 generated strongly outward-rectifying Cl(-) currents with the conductance sequence: SCN(-) >> NO(3)(-) >> Cl(-) > Br(-) approximate I(-) >> aspartate. Continuous activity of hClC-4 was sustained to different degrees by internal nucleotides: ATP approximately ATPgammaS >> AMP-PNP approximate GTP > ADP. Although non-hydrolysable nucleotides are sufficient for channel function, ATP hydrolysis is required for full activity. Changing the extracellular (2 mM or nominal Ca(2+)-free) or intracellular Ca(2+) (25 or 250 nM) concentration did not alter hClC-4 currents. Acidification of external pH (pH(o)) inhibited hClC-4 currents (half-maximal inhibition approximate 6.19), whereas neither external alkalinization to pH 8.4 nor internal acidification to pH 6.0 reduced current levels. Single-channel recordings demonstrated a Cl(-) channel active only at depolarizing potentials with a slope conductance of approximately 3 pS. Acidic pH(o) did not alter single-channel conductance. We conclude that recombinant hClC 4 encodes a small-conductance, nucleotide-dependent, Ca(2+)-independent outward rectifying chloride channel that is inhibited by external acidification. This detailed characterization will be highly valuable in comparisons of hClC-4 function with native chloride channel activities and for future structure function correlations. PMID- 11882672 TI - Inward-rectifying anion channels are expressed in the epithelial cells of choroid plexus isolated from ClC-2 'knock-out' mice. AB - Choroid plexus epithelial cells express inward-rectifying anion channels which have a high HCO(3)(-) permeability. These channels are thought to have an important role in the secretion of cerebrospinal fluid. The possible relationship between these channels and the ClC-2 Cl(-) channel was investigated in the present study. RT-PCR, using specific ClC-2 primers, amplified a 238 bp fragment of mRNA from rat choroid plexus, which was 99 % identical to the 5' sequence of rat ClC-2. A 2005 bp clone was isolated from a rat choroid plexus cDNA library using a probe for ClC-2. The clone showed greater than 99 % identity with the sequence of rat ClC-2. Inward-rectifying anion channels were observed in whole cell recordings of choroid plexus epithelial cells isolated from ClC-2 knock-out mice. The mean inward conductance was 19.6 plus minus 3.6 nS (n = 8) in controls (3 heterozygote animals), and 22.5 plus minus 3.1 nS (n = 10) in three knock-out animals. The relative permeability of the conductances to I(-) and Cl(-) (P(I) : P(Cl)) was determined. I(-) was more permeant than Cl(-) in both heterozygotes (P(I):P(Cl) = 4.0 +/- 0.9, n = 3) and knock-out animals (P(I) : P(Cl) = 4.1 +/- 1.4, n = 3). These results indicate that rat choroid plexus expresses the ClC-2 variant that was originally reported in other tissues. ClC-2 does not contribute significantly to inward-rectifying anion conductance in mouse choroid plexus, which must therefore express a novel inward-rectifying anion channel. PMID- 11882674 TI - Ca(2+)-regulated, neurosecretory granule channel involved in release from neurohypophysial terminals. AB - Ion channels from bovine neurohypophysial secretory granules (NSG) were incorporated into artificial lipid bilayers. Specific antibodies against identified synaptic vesicle proteins were tested on such incorporated channel activity and on peptide release from rat permeabilized neurohypophysial terminals. Both the NSG cation channel and Ca(2+)-dependent release were inhibited by only SY-38, a monoclonal antibody directed against the C-terminus of synaptophysin. SY-38 and Ca(2+) altered both the gating and conductance of the NSG cation channel, but in opposite ways. The close correlation between SY-38 effects on Ca(2+)-dependent channel activity and release leads us to conclude that this synaptophysin-like NSG channel is directly involved in peptide secretion from these central nervous system terminals. PMID- 11882673 TI - Changes in mechanosensitive channel gating following mechanical stimulation in skeletal muscle myotubes from the mdx mouse. AB - We studied the effects of membrane stretch and voltage on the gating of single mechanosensitive (MS) channels in myotubes from dystrophin-deficient mdx mice. In earlier studies of MS channels in mdx myotubes, we found a novel class of stretch inactivated channels. In the present experiments, we used a gentle suction protocol to determine whether seal formation damaged the membrane and altered MS channel gating, since dystrophin-deficiency is known to be associated with an increased susceptibility to mechanically induced damage. In some recordings from mdx myotubes, MS channel open probability gradually increased to levels approaching unity following seal formation. In these recordings, channels remained open for the duration of the recording. In other recordings, MS channel open probability remained low after seal formation and applying weak suction evoked conventional stretch-activated gating. Applying strong suction or very positive voltages, however, caused some channels to enter a high open probability gating mode. The shift to a high open probability gating mode coincided with the appearance of stretch-inactivated gating. These findings suggested that mechanical stimulation altered the mechanical properties of the patch causing some MS channels to enter a novel gating mode. In support of this idea, stretch activated and stretch-inactivated channels were not detected in the same membrane patch and channel inactivation occurred at lower pressures than activation (P(1/2,) = -13 and -26.5 mmHg, respectively). Other experiments showed that stretch-inactivated gating was not due to a simple loss of MS channel activity from a non-random process such as vesiculation or bleb formation: channel inactivation by suction was readily reversible, stable over tens of minutes, and followed the predictions of the binomial theorem for independent, randomly gating channels. In addition, the voltage-dependent gating of stretch-inactivated channels was similar to that of stretch-activated channels. The results show that MS channels in dystrophin-deficient muscle exist in two distinct gating modes and that mechanical stimuli cause an irreversible conversion between modes. We discuss possible mechanisms for the changes in MS channel gating in relation to the known cytoskeletal abnormalities of mdx muscle and its possible implications for the pathogenesis of Duchenne dystrophy. PMID- 11882675 TI - Differential facilitation of N- and P/Q-type calcium channels during trains of action potential-like waveforms. AB - Inhibition of presynaptic voltage-gated calcium channels by direct G-protein betagamma subunit binding is a widespread mechanism that regulates neurotransmitter release. Voltage-dependent relief of this inhibition (facilitation), most likely to be due to dissociation of the G-protein from the channel, may occur during bursts of action potentials. In this paper we compare the facilitation of N- and P/Q-type Ca(2+) channels during short trains of action potential-like waveforms (APWs) using both native channels in adrenal chromaffin cells and heterologously expressed channels in tsA201 cells. While both N- and P/Q-type Ca(2+) channels exhibit facilitation that is dependent on the frequency of the APW train, there are important quantitative differences. Approximately 20 % of the voltage-dependent inhibition of N-type I(Ca) was reversed during a train while greater than 40 % of the inhibition of P/Q-type I(Ca) was relieved. Changing the duration or amplitude of the APW dramatically affected the facilitation of N-type channels but had little effect on the facilitation of P/Q type channels. Since the ratio of N-type to P/Q-type Ca(2+) channels varies widely between synapses, differential facilitation may contribute to the fine tuning of synaptic transmission, thereby increasing the computational repertoire of neurons. PMID- 11882676 TI - Prostaglandin E(2) inhibits calcium current in two sub-populations of acutely isolated mouse trigeminal sensory neurons. AB - Prostaglandins are important mediators of pain and inflammation. We have examined the effects of prostanoids on voltage-activated calcium currents (I(Ca)) in acutely isolated mouse trigeminal sensory neurons, using standard whole cell voltage clamp techniques. Trigeminal neurons were divided into two populations based on the presence (Type 2) or absence (Type 1) of low voltage-activated T type I(Ca). The absence of T-type I(Ca) is highly correlated with sensitivity to mu-opioid agonists and the VR1 agonist capsaicin. In both populations of cells, high voltage-activated I(Ca) was inhibited by PGE(2) with an EC(50) of about 35 nM, to a maximum of 30 %. T-type I(Ca) was not inhibited by PGE(2). Pertussis toxin pre-treatment abolished the effects of PGE(2) in Type 2 cells, but not in Type 1 cells, whereas treatment with cholera toxin prevented the effects of PGE(2) in Type 1 cells, but not in Type 2 cells. Inhibition of I(Ca) by PGE(2) was associated with slowing of current activation and could be relieved with a large positive pre-pulse, consistent with inhibition of I(Ca) by G protein betagamma subunits. Reverse transcription-polymerase chain reaction of mRNA from trigeminal ganglia indicated that all four EP prostanoid receptors were present. However, in both Type 1 and Type 2 cells the effects of PGE(2) were only mimicked by the selective EP(3) receptor agonist ONO-AE-248, and not by selective agonists for EP(1) (ONO-DI-004), EP(2) (ONO-AE1-259) and EP(4) (ONO-AE1-329) receptors. These data indicate that two populations of neurons in trigeminal ganglia differing in their calcium channel expression, sensitivity to mu-opioids and capsaicin also have divergent mechanisms of PGE(2)-mediated inhibition of calcium channels, with Gi/Go type G proteins involved in one population, and Gs type G proteins in the other. PMID- 11882677 TI - Dissociation of the store-operated calcium current I(CRAC) and the Mg-nucleotide regulated metal ion current MagNuM. AB - Rat basophilic leukaemia cells (RBL-2H3-M1) were used to study the characteristics of the store-operated Ca(2+) release-activated Ca(2+) current (I(CRAC)) and the magnesium-nucleotide-regulated metal cation current (MagNuM) (which is conducted by the LTRPC7 channel). Pipette solutions containing 10 mM BAPTA and no added ATP induced both currents in the same cell, but the time to half-maximal activation for MagNuM was about two to three times slower than that of I(CRAC). Differential suppression of I(CRAC) was achieved by buffering free [Ca(2+)](i) to 90 nM and selective inhibition of MagNuM was accomplished by intracellular solutions containing 6 mM Mg.ATP, 1.2 mM free [Mg(2+)](i) or 100 microM GTP-gamma-S, allowing investigations on these currents in relative isolation. Removal of extracellular Ca(2+) and Mg(2+) caused both currents to be carried significantly by monovalent ions. In the absence or presence of free [Mg(2+)](i), I(CRAC) carried by monovalent ions inactivated more rapidly and more completely than MagNuM carried by monovalent ions. Since several studies have used divalent-free solutions on either side of the membrane to study selectivity and single-channel behaviour of I(CRAC), these experimental conditions would have favoured the contribution of MagNuM to monovalent conductance and call for caution in interpreting results where both I(CRAC) and MagNuM are activated. PMID- 11882678 TI - Skeletal muscle L-type Ca(2+) current modulation in gamma1-deficient and wildtype murine myotubes by the gamma1 subunit and cAMP. AB - Modulation of the steady-state inactivation and current amplitude by the gamma1 subunit of the murine skeletal muscle L-type Ca(2+) channel were investigated using the whole-cell patch-clamp technique. Transient expression of the gamma1 subunit, but not of the gamma2 (stargazin) protein, in primary cultured myotubes from gamma1-deficient mice shifted the steady-state inactivation approximately 15 mV, thereby restoring wildtype (WT) steady-state inactivation and current amplitude. The increased Ca(2+) current amplitude in gamma1-deficient cells was abolished in myotubes from animals of 4 weeks and older whereas the positive shift in steady-state inactivation was independent of mouse age. Raising intracellular cAMP levels using the membrane-permeant analogue 8-Br-cAMP led to an increase in Ca(2+) current amplitude in WT cells to the level in gamma1 deficient myotubes. There was no effect on the current amplitude in gamma1 deficient cells or on the steady-state inactivation in either genotype. Rp-cAMPS, a competitive inhibitor of cAMP-dependent protein kinase, had no effect on the WT Ca(2+) current amplitude and steady-state inactivation, but diminished the current amplitude in gamma1-deficient myotubes without affecting the steady-state inactivation in these cells. These data show that the increased Ca(2+) influx in myotubes lacking the gamma1 subunit, due to right-shifted steady-state inactivation and increased L-type Ca(2+) current amplitude, is determined by the gamma1 subunit. The effect on current amplitude depends on the age of the mice and its cAMP-dependent modulation appears to be controlled by the gamma1 subunit. PMID- 11882679 TI - Membrane potential bistability is controlled by the hyperpolarization-activated current I(H) in rat cerebellar Purkinje neurons in vitro. AB - We investigated the role of the hyperpolarization-activated mixed cation current, I(H), in the control of spontaneous action potential firing of rat cerebellar Purkinje neurons in brain slices. Extracellular recordings revealed that the continual action potential firing of Purkinje neurons was disrupted by the pharmacological blockade of I(H). Blockade of I(H) revealed spontaneous transitions between periods of tonic action potential firing and quiescence, without effects on the frequency or variance of action potential generation. Whole-cell recordings revealed that blockade of I(H) unmasked a form of membrane potential bistability, where transitions between tonic firing and quiescent states (separated by approximately 20 mV) were evoked by excitatory and inhibitory postsynaptic potentials, or by the delivery of brief (20 ms) somatic or dendritic positive and negative current pulses. The stable upper state of tonic action potential firing was maintained by the recruitment of axo-somatic voltage-activated sodium, but not calcium, channels. Negative modulation of I(H) by serotonin unmasked bistability, indicating that bistability of Purkinje neurons is likely to occur under physiological conditions. These data indicate that I(H) acts as a 'safety net', maintaining the membrane potential of Purkinje neurons within the range necessary for the generation of tonic action potential firing. Following the downregulation of I(H), synaptic inhibition can generate long periods (seconds) of quiescence, the duration of which can be controlled by climbing fibre activation and by the underlying 'tone' of parallel fibre activity. PMID- 11882680 TI - ASIC-like, proton-activated currents in rat hippocampal neurons. AB - The expression of mRNA for acid sensing ion channels (ASIC) subunits ASIC1a, ASIC2a and ASIC2b has been reported in hippocampal neurons, but the presence of functional hippocampal ASIC channels was never assessed. We report here the first characterization of ASIC-like currents in rat hippocampal neurons in primary culture. An extracellular pH drop induces a transient Na(+) current followed by a sustained non-selective cation current. This current is highly sensitive to pH with an activation threshold around pH 6.9 and a pH(0.5) of 6.2. About half of the total peak current is inhibited by the spider toxin PcTX1, which is specific for homomeric ASIC1a channels. The remaining PcTX1-resistant ASIC-like current is increased by 300 microM Zn(2+) and, whereas not fully activated at pH 5, it shows a pH(0.5) of 6.0 between pH 7.4 and 5. We have previously shown that Zn(2+) is a co-activator of ASIC2a-containing channels. Thus, the hippocampal transient ASIC like current appears to be generated by a mixture of homomeric ASIC1a channels and ASIC2a-containing channels, probably heteromeric ASIC1a+2a channels. The sustained non-selective current suggests the involvement of ASIC2b-containing heteromeric channels. Activation of the hippocampal ASIC-like current by a pH drop to 6.9 or 6.6 induces a transient depolarization which itself triggers an initial action potential (AP) followed by a sustained depolarization and trains of APs. Zn(2+) increases the acid sensitivity of ASIC channels, and consequently neuronal excitability. It is probably an important co-activator of ASIC channels in the central nervous system. PMID- 11882681 TI - Tyrosine kinases enhance the function of glycine receptors in rat hippocampal neurons and human alpha(1)beta glycine receptors. AB - Glycine receptors (GlyRs) are transmitter-gated channels that mediate fast inhibitory neurotransmission in the spinal cord and brain. The GlyR beta subunit contains a putative tyrosine phosphorylation site whose functional role has not been determined. To examine if protein tyrosine kinases (PTKs) regulate the function of GlyRs, we analysed whole-cell currents activated by applications of glycine to CA1 hippocampal neurons and spinal neurons. The role of a putative site for tyrosine phosphorylation at position 413 of the beta subunit was examined using site-directed mutagenesis and expression of recombinant (alpha(1)beta(Y413F)) receptors in human embryonic kidney (HEK 293) cells. Lavendustin A, an inhibitor of PTKs, depressed glycine-evoked currents (I(Gly)) in CA1 neurons and spinal neurons by 31 % and 40 %, respectively. In contrast, the intracellular application of the exogenous tyrosine kinase, cSrc, enhanced I(Gly) in CA1 neurons by 56 %. cSrc also accelerated GlyR desensitization and increased the potency of glycine 2-fold (control EC(50) = 143 microM; cSrc EC(50) = 74 microM). Exogenous cSrc, applied intracellularly, upregulated heteromeric alpha(1)beta receptors but not homomeric alpha(1) receptors. Substitution mutation of the tyrosine to phenylalanine at position beta-413 prevented this enhancement. Furthermore, a selective inhibitor of the Src family kinases, PP2, down-regulated wild-type alpha(1)beta but not alpha(1)beta(Y413F) receptors. Together, these findings indicate that GlyR function is upregulated by PTKs and this modulation is dependent on the tyrosine-413 residue of the beta subunit. PMID- 11882682 TI - Hypoxia-induced secretion of serotonin from intact pulmonary neuroepithelial bodies in neonatal rabbit. AB - We examined the effects of hypoxia on the release of serotonin (5-HT) from intact neuroepithelial body cells (NEB), presumed airway chemoreceptors, in rabbit lung slices, using amperometry with carbon fibre microelectrodes. Under normoxia (P(O2) ~155 mmHg; 1 mmHg approximately 133 Pa), most NEB cells did not exhibit detectable secretory activity; however, hypoxia elicited a dose-dependent (P(O2) range 95-18 mmHg), tetrodotoxin (TTX)-sensitive stimulation of spike-like exocytotic events, indicative of vesicular amine release. High extracellular K(+) (50 mM) induced a secretory response similar to that elicited by severe hypoxia. Exocytosis was stimulated in normoxic NEB cells after exposure to tetraethylammonium (20 mM) or 4-aminopyridine (2 mM). Hypoxia-induced secretion was abolished by the non-specific Ca(2+) channel blocker Cd(2+) (100 microM). Secretion was also largely inhibited by the L-type Ca(2+) channel blocker nifedipine (2 microM), but not by the N-type Ca(2+) channel blocker omega conotoxin GVIA (1 microM). The 5-HT(3) receptor blocker ICS 205 930 also inhibited secretion from NEB cells under hypoxia. These results suggest that hypoxia stimulates 5-HT secretion from intact NEBs via inhibition of K(+) channels, augmentation of Na(+)-dependent action potentials and calcium entry through L-type Ca(2+) channels, as well as by positive feedback activation of 5 HT(3) autoreceptors. PMID- 11882683 TI - Analysis of variance study of the rat cortical layer 4 barrel and layer 5b neurones. AB - Unique formation of rodent cortical barrels by layer 4 neurones attracts study of the sensory function of cortical input stage neurones (layer 4) compared with that of output stage neurones (layer 5). We have recorded extracellular responses from rat somatosensory cortical neurones to deflections of contralateral vibrissae. Thirty-two layer 4 barrel neurones and 29 layer 5b neurones were studied. Whisker stimulations were ramp-and-hold deflections with one of six different ramp velocities (100-2.5 mm s(-1)) and one of four different plateau amplitudes (2000-200 microm). Twenty-four (6 x 4) different stimulus forms were applied to the tip of a whisker trimmed to 10 mm in a predetermined order in stimulus cycles of 20-50 repetitions. Spike counts for a period of 2560 ms in 10 ms bins were summed to construct a matrix of 24 peristimulus histograms for each neurone. Twenty-four amplitude and 24 velocity values were computed from counts during the plateau and ramp phases, respectively. To determine the amplitude- and velocity dependence of a neurone, an amplitude F value (the ratio of variations among-/within-amplitude of the amplitude value) and a velocity F value (ratio of variations among-/within-velocity of the velocity value) were derived by analysis of variance. The amplitude F value of the layer 4 barrel neurones was greater than that of the layer 5b neurones (P < 0.0001). The velocity F value of the barrel neurones was smaller than that of the layer 5b neurones (P = 0.0226). The results suggests that barrel neurones and layer 5b neurones tend to detect amplitude and velocity components of whisker deflection, respectively. PMID- 11882684 TI - The effects of temperature on vesicular supply and release in autaptic cultures of rat and mouse hippocampal neurons. AB - Membrane fusion plays a central role in the synaptic vesicle cycle. While many of the pre- and postfusion events have been investigated at room temperature, few researchers have investigated these processes at more physiologically relevant temperatures. We have used autaptic cultures of hippocampal neurons to investigate changes in the size and refilling rate of the readily releasable pool (RRP) of synaptic vesicles brought about by an increase in temperature from 25 to 35 degrees C. We have also examined temperature-dependent changes in spontaneous and action potential (AP)-evoked release as well as the fraction of the RRP that is released during an AP. Although we found a threefold increase in the refilling rate of the RRP at the higher temperature, there was no apparent change in the size of the RRP with increased temperature. Moreover, we observed a slight but significant decrease in the quanta released during an AP. This increased refilling rate and decreased release probability resulted in a reduction of both the degree and time course of synaptic depression during high frequency stimulation at the higher temperature. This reduction in synaptic depression was accompanied by an increased maintenance of the synchronous component of release during high frequency stimulation. These findings indicate that the dynamics of vesicular supply and release in hippocampal neurons at room temperature are significantly different at near physiological temperatures and could affect our present understanding of the way in which individual neurons and networks of neurons process information. PMID- 11882685 TI - CD59 blocks not only the insertion of C9 into MAC but inhibits ion channel formation by homologous C5b-8 as well as C5b-9. AB - Activation of the complement system on the cell surface results in the insertion of pore forming membrane attack complexes (MAC, C5b-9). In order to protect themselves from the complement attack, the cells express several regulatory molecules, including the terminal complex regulator CD59 that inhibits assembly of the large MACs by inhibiting the insertion of additional C9 molecules into the C5b-9 complex. Using the whole cell patch clamp method, we were able to measure accumulation of homologous MACs in the membrane of CD59(-) human B-cells, which formed non-selective ion channels with a total conductance of 360 +/- 24 pS as measured at the beginning of the steady-state phase of the inward currents. C5b-8 and small-size MAC (MAC containing only a single C9) can also form ion channels. Nevertheless, in CD59(+) human B-cells in spite of small-size MAC formation, an ion current could not be detected. In addition, restoring CD59 to the membrane of the CD59(-) cells inhibited the serum-evoked inward current. The ion channels formed by the small-size MAC were therefore sealed, indicating that CD59 directly interfered with the pore formation of C5b-8 as well as that of small-size C5b-9. These results offer an explanation as to why CD59-expressing cells are not leaky in spite of a buildup of homologous C5b-8 and small-size MAC. Our experiments also confirmed that ion channel inhibition by CD59 is subject to homologous restriction and that CD59 cannot block the conductivity of MAC when generated by xenogenic (rabbit) serum. PMID- 11882686 TI - Muscle contraction under capillaries in hamster muscle induces arteriolar dilatation via K(ATP) channels and nitric oxide. AB - We tested the hypothesis that adenosine and nitric oxide can be sensed by capillaries and are implicated in the remote arteriolar dilatation initiated by muscle contraction. We also explored a role for K(ATP) channel activity in this response. Small bundles of muscle fibres underlying a group of capillaries in cremaster muscles of anaesthetized hamsters were electrically stimulated to contract for 2 min at each of 2, 4 and 8 Hz. Diameter changes were measured in the inflow arteriole to the group of capillaries after muscle contraction in the presence or absence of 10(-6) M xanthine amine congener (XAC) to block A(1) and A(2) adenosine receptors, 10(-4) or 10(-3) M N(omega)-nitro-L-arginine (LNNA) to block nitric oxide production, or 10(-5) M glibenclamide to block K(ATP) channel activity. Dilatations were unchanged with XAC (3.0 +/- 0.5, 3.9 +/- 0.7 and 6.1 +/- 1.0 microm), but were significantly reduced with LNNA (to 1.8 +/- 0.6, 3.5 +/ 0.7 and 4.9 +/- 0.7 microm) or glibenclamide (to 0.4 +/- 0.3, 0.8 +/- 0.7 and 1.9 +/- 0.6 microm). Neither K(ATP) channel activity nor nitric oxide was required for transmission or manifestation of the dilator response. Thus, muscle contraction can be sensed by capillaries and the signalling mechanism for the ensuing remote dilatation depends on K(ATP) channel activity and on NO, but not adenosine. Local application of 10(-4) M adenosine, 10(-4) M sodium nitroprusside or 10(-5) M pinacidil directly to capillaries initiated remote arteriolar dilatations. Thus, capillaries can respond directly to known mediators of metabolic vasodilatation, but these signalling pathways are not invariably implicated in the response to muscle contraction. PMID- 11882688 TI - Different temporal processing of sensory inputs in the rat thalamus during quiescent and information processing states in vivo. AB - Sensory inputs from the whiskers reach the primary somatosensory thalamus through the medial lemniscus tract. The main role of the thalamus is to relay these sensory inputs to the neocortex according to the regulations dictated by behavioural state. Intracellular recordings in urethane-anaesthetized rats show that whisker stimulation evokes EPSP-IPSP sequences in thalamic neurons. Both EPSPs and IPSPs depress with repetitive whisker stimulation at frequencies above 2 Hz. Single-unit recordings reveal that during quiescent states thalamic responses to repetitive whisker stimulation are suppressed at frequencies above 2 Hz, so that only low-frequency sensory stimulation is relayed to the neocortex. In contrast, during activated states, induced by stimulation of the brainstem reticular formation or application of acetylcholine in the thalamus, high frequency whisker stimulation at up to 40 Hz is relayed to the neocortex. Sensory suppression is caused by the depression of lemniscal EPSPs in relatively hyperpolarized thalamocortical neurons. Sensory suppression is abolished during activated states because thalamocortical neurons depolarize and the depressed lemniscal EPSPs are able to reach firing threshold. Strong IPSPs may also contribute to sensory suppression by hyperpolarizing thalamocortical neurons, but during activated states IPSPs are strongly reduced altogether. The results indicate that the synaptic depression of lemniscal EPSPs and the level of depolarization of thalamocortical neurons work together in thalamic primary sensory pathways to suppress high-frequency sensory inputs during non-activated (quiescent) states while permitting the faithful relay of high-frequency sensory information during activated (processing) states. PMID- 11882687 TI - Rapid actions of 17beta-oestradiol on a subset of lactotrophs in the rat pituitary. AB - Increasingly the role of rapid mechanisms of steroid action in physiological regulation are being recognised. We have investigated rapid effects of 17beta oestradiol (E) on prolactin (PRL) release in vitro. Pituitary segments from male rats were incubated for 5, 10 or 20 min in Earle's balanced salt solution containing 1.2 mM tannic acid (to enable visualisation of exocytosed secretory granules by electron microscopy) either alone (control) or containing 10(-10)-10( 8) M E conjugated to bovine serum albumin (E-BSA). PRL and leuteinising hormone (LH) release from pituitary segments were also determined in response to E and E BSA by radioimmunoassay. Within 10 min E-BSA and E (10(-12)-10(-6) M) stimulated a significant (P < 0.05) concentration-dependent release of PRL but not LH. After exposure to experimental media for 5 min, only occasional exocytosis from type I lactotrophs (characterised by large polymorphic secretory granules) was observed in either control or E-BSA treated tissue. In contrast, E-BSA (10(-10)-10(-8) M) induced a significant (P < 0.05) increase in the number of exocytotic profiles from type II lactotrophs (characterized by smaller, spherical granules). This effect was not inhibited by removal of extracellular calcium, or by pre-treatment of cells with the RNA synthesis inhibitor actinomycin-D (0.5 microg ml(-1)), the protein synthesis inhibitor cycloheximide (1 microg ml(-1)) or the anti-oestrogen ICI 182,780 (1 microM). FACS analysis demonstrated binding of E-BSA-fluorescein isothiocyanate (FITC) (10(-10)-10(-7) M) to a subpopulation of anterior pituitary cells. The E-BSA-FITC binding sites assumed a patchy distribution across the cell surface. In conclusion, we report for the first time a rapid, non-genomic effect of E on PRL secretion in normal pituitary tissue. PMID- 11882689 TI - Pathophysiological basis of acute inflammatory hyperaemia in the rat knee: roles of cyclo-oxygenase-1 and -2. AB - The role of different isoforms of cyclo-oxygenase (COX) in mediating the acute (0 6 h) and late (24 h) phases of inflammation was investigated in the rat knee joint following intra-articular injection of carrageenan. The hyperaemic response was assessed transcutaneously using laser Doppler imaging (LDI). Samples were taken at corresponding time points for detection of synovial COX-1, COX-2 and inducible nitric oxide synthase (iNOS) mRNA, and measurement of urinary prostaglandin (PG) and nitric oxide metabolites (NO(x)). A non-selective COX inhibitor (indomethacin, 15 mg kg(-1) I.P.), a selective COX-2 inhibitor (SC-236, 16.8 mg kg(-1) I.P.) or vehicle were administered 1 h prior to carrageenan in the acute phase study. LDI scans were taken hourly for 4 h post-induction. Inflammatory hyperaemia in the vehicle group was attenuated in the indomethacin- (P < 0.001, two-way ANOVA) and SC-236-treated groups (P < 0.0001), with no difference between these treatments. At 24 h, I.V. infusion of indomethacin (0.1 mg min(-1)), increased vascular resistance (24 +/- 7.1 %; P < 0.05) compared to vehicle infusion, whereas SC-236 (0.11 mg min(-1)) did not. Resistance changes to indomethacin also differed from SC-236 (P < 0.05). Knee joint diameter progressively increased over 24 h (P < 0.0001, one-way ANOVA). Urinary PG levels increased by 6 h (P < 0.05), but returned to baseline by 24 h. COX-1 mRNA was detectable at all time points; COX-2 mRNA only at 3 h. Urinary NO(x) levels increased progressively over 24 h (P < 0.05), paralleled by induction of iNOS in the 3 and 24 h samples. Prostaglandin production via COX-2 appears to mediate the development of acute inflammatory hyperaemia, but nitrergic mechanisms may supervene subsequently. COX-1 but not COX-2 contributes to the maintenance of basal blood flow in the hyperaemic joint at 24 h. PMID- 11882690 TI - Slow excitatory synaptic potentials evoked by distension in myenteric descending interneurones of guinea-pig ileum. AB - The functional significance of the slow excitatory synaptic potentials (EPSPs) in myenteric neurones is unknown. We investigated this using intracellular recording from myenteric neurones in guinea-pig ileum, in vitro. In all, 121 neurones responded with fast EPSPs to distension of the intestine oral to the recording site. In 28 of these neurones, distension also evoked depolarizations similar to the slow EPSPs evoked by electrical stimulation in the same neurones. Intracellular injection of biocytin and immunohistochemistry revealed that neurones responding to distension with slow EPSPs were descending interneurones, which were immunoreactive for nitric oxide synthase (NOS). Other neurones, including inhibitory motor neurones and interneurones lacking NOS, did not respond to distension with slow EPSPs, but many had slow EPSPs evoked electrically. Slow EPSPs evoked electrically or by distension in NOS immunoreactive descending interneurones were resistant to blockade of NK(1) or NK(3) tachykinin receptors (SR 140333, 100 nM; SR 142801, 100 nM, respectively) and group I metabotropic glutamate receptors (PHCCC, 10-30 microM), when the antagonists were applied in the recording chamber of a two-chambered organ bath. However, slow EPSPs evoked electrically in inhibitory motor neurones were substantially depressed by SR 140333 (100 nM). Blockade of synaptic transmission in the stimulation chamber of the organ bath abolished slow EPSPs evoked by distension, indicating that they arose from activity in interneurones, and not from anally directed, intrinsic sensory neurones. Thus, distension evokes slow EPSPs in a subset of myenteric neurones, which may be important for intestinal motility. PMID- 11882691 TI - Feedback inhibition in the inner plexiform layer underlies the surround-mediated responses of AII amacrine cells in the mammalian retina. AB - Intracellular recordings were made from narrow-field, bistratified AII amacrine cells in the isolated, superfused retina-eyecup of the rabbit. Pharmacological agents were applied to neurons to dissect the synaptic pathways subserving AII cells so as to determine the circuitry generating their off-surround responses. Application of the GABA antagonists, picrotoxin, bicuculline and 1,2,5,6 tetrahydropyridine-4-yl methylphosphinic acid (TPMPA) all increased the on-centre responses of AII amacrine cells, but attenuated the off-surround activity. At equal concentrations, picrotoxin was approximately twice as effective as bicuculline or TPMPA in modifying the response activity of AII amacrine cells. These results indicate that the mechanism underlying surround inhibition of AII amacrine cells includes activation of both GABA(A) and GABA(C) receptors in an approximately equal ratio. Application of the GABA antagonists also increased the size of on-centre receptive fields of AII amacrine cells. Again, picrotoxin was most effective, producing, on average, a 54 % increase in the size of the receptive field, whereas bicuculline and TPMPA produced comparable 34 and 33 % increases, respectfully. Application of the voltage-gated sodium channel blocker TTX produced effects on AII amacrine cells qualitatively similar to those of the GABA blockers. Intracellular application of the chloride channel blocker 4,4' dinitro-stilbene-2,2'-disulphonic acid (DNDS) abolished the direct effects of GABA on AII amacrine cells. Moreover, DNDS increased the amplitude of both the on centre and off-surround responses. The failure of DNDS to block the off-surround activity indicates that it is not mediated by direct GABAergic inhibition. Taken together, our results suggest that surround receptive fields of AII amacrine cells are generated indirectly by the GABAergic, reciprocal feedback synapses from S1/S2 amacrine cells to the axon terminals of rod bipolar cells. PMID- 11882692 TI - Electrical stimulation of the midbrain increases heart rate and arterial blood pressure in awake humans. AB - Electrical stimulation of the hypothalamus, basal ganglia or pedunculopontine nucleus in decorticate animals results in locomotion and a cardiorespiratory response resembling that seen during exercise. This has led to the hypothesis that parallel activation of cardiorespiratory and locomotor systems from the midbrain could form part of the 'central command' mechanism of exercise. However, the degree to which subcortical structures play a role in cardiovascular activation in awake humans has not been established. We studied the effects on heart rate (HR) and mean arterial blood pressure (MAP) of electrically stimulating the thalamus and basal ganglia in awake humans undergoing neurosurgery for movement disorders (n = 13 Parkinson's disease, n = 1 myoclonic dystonia, n = 1 spasmodic torticollis). HR and MAP increased during high frequency (> 90 Hz) electrical stimulation of the thalamus (HR 5 +/- 3 beats min( 1), P = 0.002, MAP 4 +/- 3 mmHg, P = 0.05, n = 9), subthalamic nucleus (HR 5 +/- 3 beats min(-1), P = 0.002, MAP 5 +/- 3 mmHg, P = 0.006, n = 8) or substantia nigra (HR 6 +/- 3 beats min(-1), P = 0.001, MAP 5 +/- 2 mmHg, P = 0.005, n = 8). This was accompanied by the facilitation of movement, but without the movement itself. Stimulation of the internal globus pallidus did not increase cardiovascular variables but did facilitate movement. Low frequency (< 20 Hz) stimulation of any site did not affect cardiovascular variables or movement. Electrical stimulation of the midbrain in awake humans can cause a modest increase in cardiovascular variables that is not dependent on movement feedback from exercising muscles. The relationship between this type of response and that occurring during actual exercise is unclear, but it indicates that subcortical command could be involved in 'parallel activation' of the locomotor and cardiovascular systems and thus contribute to the neurocircuitry of 'central command'. PMID- 11882693 TI - Detailed passive cable models of layer 2/3 pyramidal cells in rat visual cortex at different temperatures. AB - We present detailed passive cable models of layer 2/3 pyramidal cells based on somatic voltage transients in response to brief current pulses at physiological and room temperatures and demonstrate how cooling alters the shape of postsynaptic responses. Whole cell recordings were made from cells in visual cortical slices from 20- to 22-day-old rats. The cells were filled with biocytin and morphologies were reconstructed from three cells which were representative of the full range of physiological responses. These formed the basis for electrotonic models with four electrical variables, namely membrane capacitance (C(m)), membrane resistivity (R(m)), cytoplasmic resistivity (R(i)) and a somatic shunt conductance (G(sh)). Simpler models, with a single value for R(m) and no G(sh), did not fit the data adequately. Optimal parameter values were derived by simulating the responses to somatic current pulses, varying the parameters to give the best match to the experimental recordings. G(sh) and R(m) were badly constrained. In contrast, the total membrane conductance (G(tot)) was well constrained, and its reciprocal correlated closely with the slowest membrane time constant (tau(0)). The models showed close agreement for C(m) and R(i) (ranges at 36 degrees C: 0.78-0.94 microF cm(-2) and 140-170 Omegacm), but a larger range for G(tot) (7.2-18.4 nS). Cooling produced consistent effects in all three model cells; C(m) remained constant (Q(10) = 0.96), R(i) increased (Q(10) = 0.80), whilst G(tot) dropped (Q(10) = 1.98). In terms of whole cell physiology, the predominant effect of cooling is to dramatically lengthen the decay of transient voltage shifts. Simulations suggest that this markedly increases the temporal summation of postsynaptic potentials and we demonstrate this effect in the responses of layer 2/3 cells to tetanic extracellular stimulation in layer 4. PMID- 11882695 TI - In situ protection of microbiodiversity is under consideration. PMID- 11882694 TI - Change in length of relaxed muscle fascicles and tendons with knee and ankle movement in humans. AB - Ultrasonography was used to measure changes in length of muscle fascicles in relaxed human tibialis anterior and gastrocnemius during passively imposed changes in joint angle. Changes in the length of muscle fascicles were compared to changes in the length of the whole muscle-tendon units calculated from joint angles and anthropometric data. Relaxed muscle fascicles underwent much smaller changes in length than their muscle-tendon units. On average, muscle fascicles in tibialis anterior 'saw' 55 +/- 13 % (mean +/- S.D.) of the total change in muscle tendon length. This indicates nearly half of the total change in muscle-tendon length was taken up by stretch of tendon. In gastrocnemius, which has relatively long tendons, only 27 +/- 9 % of the total change in muscle-tendon length was transmitted to muscle fascicles. Thus, the tendency for passive movement to be taken up by the tendon was greater for gastrocnemius than tibialis anterior (P = 0.002). For these muscles, the relatively large changes in tendon length across much of the physiological range of muscle-tendon lengths could not wholly be explained by tendon slackness, changes in fibre pennation, or stretch or contraction history of the muscle. Our data confirm that when joints are moved passively, length changes 'seen' by muscle fascicles can be much less than changes in the distance between muscle origin and insertion. This occurs because tendons undergo significant changes in length, even at very low forces. PMID- 11882696 TI - Sinorhizobial chemotaxis: a departure from the enterobacterial paradigm. PMID- 11882697 TI - Intragenomic and intraspecific polymorphism of the 16S-23S rDNA internally transcribed sequences of Streptomyces ambofaciens. AB - The nucleotide composition of the internally transcribed sequences (ITSs) of the six rDNA operons of two strains of Streptomyces ambofaciens were determined. Four variable and five conserved nucleotide blocks were distinguished. Five different modular organizations were revealed for each strain and no homologous loci showed the same succession of blocks. This suggests that recombination frequently occurs between the rDNA loci, leading to the exchange of nucleotide blocks. The modular structure was also observed within the ITSs of Streptomyces coelicolor M145, which is closely related to Streptomyces ambofaciens, and Streptomyces griseus 2247, showing the same number of constant blocks but with fewer variable regions. This confirms that a high degree of ITS variability is a common characteristic among Streptomyces spp. The functional significance of the combinations of variable and constant nucleotide blocks of the ITS was examined by in silico prediction of secondary structures from nucleotide sequences. The secondary structures were shown to be analogous whatever the combination of variable/constant blocks at the intragenomic, intraspecific and interspecific levels. PMID- 11882698 TI - The positive activator of cephamycin C and clavulanic acid production in Streptomyces clavuligerus is mistranslated in a bldA mutant. AB - In Streptomyces coelicolor bldA encodes the principal leucyl tRNA for translation of UUA codons and controls pigmented antibiotic production by the presence of TTA codons in the genes encoding the pathway-specific activators of actinorhodin and undecylprodigiosin biosynthesis. In Streptomyces clavuligerus the gene encoding the pathway-specific activator of both cephamycin C and clavulanic acid production, ccaR, also contains a TTA codon and was expected to exhibit bldA dependence. A cloned S. clavuligerus DNA fragment containing a sequence showing 91% identity to the S. coelicolor bldA-encoded tRNA was able to restore antibiotic production and sporulation to bldA mutants of S. coelicolor and the closely related Streptomyces lividans. A null mutation of the bldA gene in S. clavuligerus resulted in the expected sporulation defective phenotype, but unexpectedly had no effect on antibiotic production. Transcript analysis showed no difference in the levels of ccaR transcripts in the wild-type and bldA mutant strains, ruling out any effect of elevated levels of the ccaR mRNA. Furthermore, when compared to the wild-type strain, the bldA mutant showed no differences in the levels of CcaR, suggesting that the single TTA codon in ccaR is mistranslated efficiently. The role of codon context in bldA dependence is discussed. PMID- 11882699 TI - Mutagenesis of conserved tryptophan residues within the receptor-binding domain of intimin: influence on binding activity and virulence. AB - Intimate bacterial adhesion to intestinal epithelium is a pathogenic mechanism shared by several human and animal enteric pathogens, including enteropathogenic and enterohaemorrhagic Escherichia coli and Citrobacter rodentium. The proteins directly involved in this process are the outer-membrane adhesion molecule intimin and the translocated intimin receptor, Tir. The receptor-binding activity of intimin resides within the carboxy terminus 280 aa (Int280) of the polypeptide. Four tryptophan residues, W117/776, W136/795, W222/881 and W240/899, are conserved within different Int280 molecules that otherwise show considerable sequence variation. In this study the influence of site-directed mutagenesis of each of the four tryptophan residues on intimin-Tir interactions and on intimin mediated intimate attachment was determined. The mutant intimins were also studied using a variety of in vitro and in vivo infection models. The results show that all the substitutions modulated intimin activity, although some mutations had more profound effects than others. PMID- 11882700 TI - Contribution of the ADP-ribosylating and receptor-binding properties of cholera like enterotoxins in modulating cytokine secretion by human intestinal epithelial cells. AB - When epithelial cells first encounter cholera toxin (Ctx) produced by Vibrio cholerae they secrete not only chloride ions responsible for causing diarrhoea, but also a number of cytokines that may contribute to the toxin's potent immunomodulatory properties. Much less is known about the ability of the heat labile enterotoxin of Escherichia coli (Etx), a close homologue of Ctx, to elicit cytokine secretion by epithelial cells. This study shows that treatment of human intestinal epithelial T84 cells with Etx induces expression of IL-6, IL-10, IL-1R antagonist, as well as IL-1alpha and IL-1beta and low levels of IL-8. Such induction was totally dependent on the intrinsic ADP-ribosylating activity of the toxin A-subunit, and could be mimicked by cAMP-elevating agents, such as forskolin and dibutyryl cAMP. By comparison, neither an enzymically inactive mutant of Etx nor EtxB was able to induce cytokine secretion. The behaviour of Ctx and CtxB was very similar to that of Etx and EtxB, respectively. The spectrum of cytokines released by Etx and Ctx indicates that the toxins may create a local microenvironment that strongly biases the immune response towards an anti inflammatory and a polarized Th2 response. PMID- 11882701 TI - Influence of activated charcoal, porcine gastric mucin and beta-cyclodextrin on the morphology and growth of intestinal and gastric Helicobacter spp. AB - Bile-tolerant Helicobacter spp. are emerging human and animal pathogens. However, due to their fastidious nature, which requires nutrient-rich complex media to grow, infection with these bacteria may be underestimated. The accumulation of toxic metabolites in cultures may be one of the main obstacles for successful culture of these organisms. The present study examined various potential growth enhancing substances for Helicobacter spp. and, furthermore, how they may affect spiral to coccoid conversion. Five Helicobacter spp. were cultured on agar and in broth media supplemented with activated charcoal, beta-cyclodextrin, or porcine gastric mucin. Growth was determined by estimating the numbers of colony-forming units and colony diameter, as well as bacterial cell mass. Coccoid transformation was estimated every 24 h by both Gram and acridine-orange staining. Activated charcoal was superior in supporting growth and increased cell mass on agar and in broth media. beta-Cyclodextrin delayed spiral to coccoid conversion by Helicobacter pylori and Helicobacter canis, whereas activated charcoal delayed the conversion to coccoid forms of Helicobacter hepaticus and Helicobacter bilis. The progression to coccoid forms by Helicobacter pullorum on agar media was not influenced by any growth supplement. The spiral to coccoid conversion was more rapid in broth media than on agar media. The growth enhancement observed is probably related to the capacity of activated charcoal to remove toxic compounds in culture media. PMID- 11882702 TI - Analysis of gluconeogenic and anaplerotic enzymes in Campylobacter jejuni: an essential role for phosphoenolpyruvate carboxykinase. AB - Campylobacter jejuni is unable to utilize glucose as a carbon source due to the absence of the key glycolytic enzyme 6-phosphofructokinase. The genome sequence of C. jejuni NCTC 11168 indicates that homologues of all the appropriate enzymes for gluconeogenesis from phosphoenolpyruvate (PEP) are present, in addition to the anaplerotic enzymes pyruvate carboxylase (PYC), phosphoenolpyruvate carboxykinase (PCK) and malic enzyme (MEZ). Surprisingly, a pyruvate kinase (PYK) homologue is also present. To ascertain the role of these enzymes, insertion mutants in pycA, pycB, pyk and mez were generated. However, this could not be achieved for pckA, indicating that PCK is an essential enzyme in C. jejuni. The lack of PEP synthase and pyruvate orthophosphate dikinase activities confirmed a unique role for PCK in PEP synthesis. The pycA mutant was unable to grow in defined medium with pyruvate or lactate as the major carbon source, thus indicating an important role for PYC in anaplerosis. Sequence and biochemical data indicate that the PYC of C. jejuni is a member of the alpha4beta4, acetyl CoA-independent class of PYCs, with a 65.8 kDa subunit containing the biotin moiety. Whereas growth of the mez mutant was comparable to that of the wild-type, the pyk mutant displayed a decreased growth rate in complex medium. Nevertheless, the mez and pyk mutants were able to grow with pyruvate, lactate or malate as carbon sources in defined medium. PYK was present in cell extracts at a much higher specific activity [>800 nmol x min(-1) x (mg protein)(-1)] than PYC or PCK [<65 nmol x min(-1) x (mg protein)(-1)], was activated by fructose 1,6 bisphosphate and displayed other regulatory properties strongly indicative of a catabolic role. It is concluded that PYK may function in the catabolism of unidentified substrates which are metabolized through PEP. In view of the high K(m) of MEZ for malate (approximately 9 mM) and the lack of a growth phenotype of the mez mutant, MEZ seems to have only a minor anaplerotic role in C. jejuni. PMID- 11882703 TI - In vivo effect of mutations in the antiterminator LacT in Lactobacillus casei. AB - The antiterminator LacT regulates the expression of the lactose operon in Lactobacillus casei and its activity is controlled by EII(Lac) and common PTS elements. LacT shows the two conserved domains (PRD-I and PRD-II) characteristic of the BglG antiterminator family that are implicated in the regulation of their activity, possibly by phosphorylation of conserved histidines. By site-directed mutagenesis of LacT, four histidines (His-101, His-159 in PRD-I and His-210, His 273 in PRD-II) were replaced by alanine or aspartate, mimicking non phosphorylated and phosphorylated forms, respectively. These constructions were used to complement DeltalacT and DeltaccpA mutants. L. casei strains (DeltalacT) carrying the replacement of His-101 or His-159 by Ala showed phospho-beta galactosidase activity in absence of the inducer (lactose), indicating that these amino acids, located in PRD-I, are essential for EII-dependent induction of the lac operon, possibly by dephosphorylation. Interestingly, these mutations rendered LacT thermosensitive. Moreover, expression of H210A and H273A (PRD-II) mutations in L. casei DeltaccpA showed that these two histidyl residues could have a role in LacT-dependent carbon catabolite repression (CCR) of this system. Overexpression of LacT in a ccpA background rendered the lac operon insensitive to CCR, but it was still sensitive to lactose induction. This suggests that the transfer of phosphate groups from PTS elements, which controls these two regulatory processes (CCR and substrate induction), could have different affinity for PRD-I and PRD-II histidines. PMID- 11882704 TI - The Enterococcus faecalis gene encoding the novel general stress protein Gsp62. AB - The Enterococcus faecalis general stress protein Gsp62 was purified using two dimensional gel electrophoresis and its 25 N-terminal amino acid sequence determined. Analysis of the corresponding gene revealed that the gsp62 product is a 172 aa protein. Transcriptional analysis of gsp62 gave evidence for a monocistronic mRNA, the synthesis of which was induced at the onset of stationary phase and in response to heat shock, acid pH, detergents (i.e. SDS or bile salts), ethanol, tert-butyl hydroperoxide, sodium chloride and, to a lesser extent, hydrogen peroxide. 5' rapid amplification of cDNA ends by PCR experiments showed that gsp62 transcription initiates 30 nt upstream of the ATG start codon. Although gsp62 expression was induced in response to various stresses, its disruption had no significant effect on the cell survival after each individual stress. Two-dimensional protein gels from wild-type and mutant cells revealed no pleiotropic effect of the mutation on protein synthesis. Transcriptional fusions with the lacL lacM beta-galactosidase genes showed that an inverted repeat located upstream of the promoter is required for transcriptional induction by environmental stresses but not by entrance into stationary phase. Two distinct mechanisms responding to different signals are thus involved in gsp62 induction. PMID- 11882705 TI - Isoleucine starvation caused by sulfometuron methyl in Salmonella typhimurium measured by translational frameshifting. AB - The authors have developed a tool for the study of inhibitor-induced amino acid starvation in bacteria which exploits the phenomenon of translational frameshifting. The inhibition of acetohydroxyacid synthase II by the herbicide sulfometuron methyl (SMM) has complex effects on branched-chain amino acid biosynthesis. Experiments were done with Salmonella typhimurium containing a plasmid with an isoleucine codon in a 'shifty' region, prone to translational frameshifting. SMM did not cause translational frameshifting in minimal medium under conditions that inhibit growth. A 20-fold higher concentration of SMM was required to cause starvation for isoleucine, e.g. in the presence of valine. This starvation was reflected in translational frameshifting correlated with inhibition of growth. These observations support the authors' previous suggestions based on other techniques. The method used here could be generalized for the study of complex metabolic effects related to amino acids. PMID- 11882706 TI - The role of two periplasmic copper- and zinc-cofactored superoxide dismutases in the virulence of Salmonella choleraesuis. AB - Periplasmic copper- and zinc-cofactored superoxide dismutases ([Cu,Zn]-SODs, SodC) of several Gram-negative pathogens can protect against superoxide-radical mediated host defences, and thus contribute to virulence. This role has been previously defined for one [Cu,Zn]-SOD in various Salmonella serovars. Following the recent discovery of a second periplasmic [Cu,Zn]-SOD in Salmonella, the effect of knockout mutations in one or both of the original sodC-1 and the new sodC-2 on the virulence of the porcine pathogen Salmonella choleraesuis is investigated here. In comparison to wild-type, while sodC mutants--whether single or double--showed no impairment in growth, they all showed equally enhanced sensitivity to superoxide and a dramatically increased sensitivity to the combination of superoxide and nitric oxide in vitro. This observation had its correlate in experimental infection both ex vivo and in vivo. Mutation of sodC significantly impaired survival of S. choleraesuis in interferon gamma-stimulated murine macrophages compared to wild-type organisms, and all S. choleraesuis sodC mutants persisted in significantly lower numbers than wild-type in BALB/c (Ity(s)) and C3H/HeN (Ity(r)) mice after experimental infection, but in no experimental system were sodC-1 sodC-2 double mutants more attenuated than either single mutant. These data suggest that both [Cu,Zn]-SODs are needed to protect bacterial periplasmic or membrane components. While SodC plays a role in S. choleraesuis virulence, the data presented here suggest that this is through overcoming a threshold effect, probably achieved by acquisition of sodC-1 on a bacteriophage. Loss of either sodC gene confers maximum vulnerability to superoxide on S. choleraesuis. PMID- 11882707 TI - Giardia intestinalis, a eukaryote without hydrogenosomes, produces hydrogen. AB - The microaerophilic flagellated protist Giardia intestinalis, the commonest protozoal agent of intestinal infections worldwide, is of uncertain phylogeny, but is usually regarded as the earliest branching of the eukaryotic clades. Under strictly anaerobic conditions, a mass spectrometric investigation of gas production indicated a low level of generation of dihydrogen (2 nmol x min(-1) per 10(7) organisms), about 10-fold lower than that in Trichomonas vaginalis under similar conditions. Hydrogen evolution was O2 sensitive, and inhibited by 100 microM metronidazole. Fluorescent labelling of G. intestinalis cells using monoclonal antibodies to typical hydrogenosomal enzymes from T. vaginalis (malate enzyme, and succinyl-CoA synthetase alpha and beta subunits), and to the large granule fraction (hydrogenosome-enriched, also from T. vaginalis) gave no discrete localization of epitopes. Cell-free extracts prepared under anaerobic conditions showed the presence of a CO-sensitive hydrogenase activity. This first report of hydrogen production in a eukaryote with no recognizable hydrogenosomes raises further questions about the early branching status of G. intestinalis; the physiological characterization of its hydrogenase, and its recently elucidated gene sequence, will aid further phylogenetic investigations. PMID- 11882708 TI - The growth cycle of Simkania negevensis. AB - Simkania negevensis, a bacterium formerly referred to as 'the micro-organism Z' or 'Simkania Z', belongs to the order Chlamydiales, assigned to the family Simkaniaceae: The purpose of this study was to investigate the production of Simkania negevensis progeny in infected cells in comparison with the well documented Chlamydiaceae developmental cycle. It was found that replicating Simkania negevensis in Vero cells resembled the reticulate bodies of all known chlamydial species: in electron micrographs they were reticulated, homogeneously staining, and often caught in the process of binary division. These replicative forms were found in low abundance shortly after infection, but by 3 days post infection they were the most prevalent particles in host cells. Electron-dense forms of Simkania negevensis began to appear on the third day post-infection, but quantitatively did not account for the high titre of infectivity in extracts from these host cells. These had both electron-dense and electron-lucent areas, a characteristic seen only in a few chlamydial species. Simkania negevensis infectivity did not appreciably change during the ensuing 12 days required for host cell lysis, despite an eightfold increase in the proportion of electron dense bacteria over this time. The emergence of electron-dense bodies, increase in infectivity and host-cell lysis were not synchronized developmental events. This is a novel finding in Chlamydiales spp. and suggests that Simkania negevensis will provide new perspectives in the mechanisms of chlamydial intracellular growth. PMID- 11882709 TI - Shell disease syndrome in the edible crab, Cancer pagurus--isolation, characterization and pathogenicity of chitinolytic bacteria. AB - Chitinolytic bacteria are believed to be the primary aetiological agents of shell disease syndrome in marine crustaceans. The disease principally results from the breakdown of their chitinous exoskeletons by the shell disease pathogens, but pathogenicity may also manifest internally should a breach of the carapace occur. The current study looks at the pathogenicity of a number of bacteria (predominantly from the genus Vibrio) isolated from the edible crab, Cancer pagurus. All chitinase-producing bacteria investigated were capable of growth in a minimal medium consisting of chitin powder from crab shells, but differed in their speed of growth and nature of chitinolytic activity, suggesting that they have different roles within the lesion community. Two isolates (designated I4 and I7) were chosen for studies on internal pathogenicity, which included the effect of the pathogen on crab tissues, the ability of the host to remove the bacteria from circulation and the antibacterial activity of crab blood. Initially, I4 was rapidly removed from circulation, but began to reappear in the blood after 24 h. By 100 h, 100% of crabs were moribund. The septicaemic effects of the isolate were reflected in the low levels of its killing by blood-cell lysate and serum. By contrast, I7 was only slowly removed from circulation and caused the rapid mortality of all crabs in <3 h. A large decline in the number of circulating blood cells following injection of I7 was mirrored by an accumulation of these cells in the gills. Initial experiments suggest that the death of the crabs following injection with I7 may be caused by toxic extracellular bacterial products that exert their effects on the blood cells and nervous system of the crabs. PMID- 11882710 TI - Evidence that ORF3 at the Streptococcus parasanguis fimA locus encodes a thiol specific antioxidant. AB - Streptococcus parasanguis is a primary colonizer of dental plaque and a major player in subacute bacterial endocarditis. In the present study, the authors report that an ORF (ORF3) located 77 bp downstream of the fimA operon on the S. parasanguis FW213 chromosome complements an Escherichia coli thiol peroxidase (tpx) mutation in glutamine synthetase (GS) protection assays and that GS is protected by the ORF3 gene product in S. parasanguis cell extracts. In addition, the putative streptococcal peroxidase (Tpx(Sp)) protects S. parasanguis from stress caused by H2O2 and is induced by oxygen, as revealed by Northern blot analysis. Taken collectively, these findings support a thiol-dependent antioxidant activity for Tpx in S. parasanguis. PMID- 11882711 TI - LuxS-dependent quorum sensing in Porphyromonas gingivalis modulates protease and haemagglutinin activities but is not essential for virulence. AB - Porphyromonas gingivalis is a Gram-negative black-pigmented obligate anaerobe implicated in the aetiology of human periodontal disease. The virulence of P. gingivalis is associated with the elaboration of the cysteine proteases Arg gingipain (Rgp) and Lys-gingipain (Kgp), which are produced at high bacterial cell densities. To determine whether quorum sensing plays a role in the regulation of Rgp and Kgp, biosensors capable of detecting either N acylhomoserine lactone (AHLs) or the luxS-dependent autoinducer (AI-2) quorum sensing signalling molecules in spent culture supernatants were first employed. While no AHLs could be detected, the Vibrio harveyi BB170 biosensor was activated by spent P. gingivalis W50 culture supernatants. The P. gingivalis luxS gene was cloned and demonstrated to restore AI-2 production in the Escherichia coli luxS mutant DH5alpha. Mutation of luxS abolished AI-2 production in P. gingivalis. Western blotting using antibodies raised against the recombinant protein revealed that LuxS levels increased throughout growth even though AI-2 activity was only maximally detected at the mid-exponential phase of growth and disappeared by the onset of stationary phase. Similar results were obtained with E. coli DH5alpha transformed with luxS, suggesting that AI-2 production is not limited by a lack of LuxS protein. Analysis of Rgp and Kgp protease activities revealed that the P. gingivalis luxS mutant produced around 45% less Rgp and 30% less Kgp activity than the parent strain. In addition, the luxS mutant exhibited a fourfold reduction in haemagglutinin titre. However, these reductions in virulence determinant levels were insufficient to attenuate the luxS mutant in a murine lesion model of P. gingivalis infection. PMID- 11882712 TI - Detection and mutation of a luxS-encoded autoinducer in Proteus mirabilis. AB - Quorum sensing regulates the expression of virulence factors in a wide variety of pathogenic bacteria. This study has shown that Proteus mirabilis harbours a homologue of luxS, a gene required for the synthesis of the quorum sensing autoinducer 2 (AI-2). AI-2 activity is expressed during and is correlated with the initiation of swarming migration on agar surfaces. The P. mirabilis luxS locus was cloned and a LuxS(-) strain constructed by allelic-exchange mutagenesis. While lacking AI-2 activity, a null mutation in luxS, however, did not affect swimming or swarming motility, swarmer cell differentiation, or virulence in a mouse model of ascending urinary tract infection. PMID- 11882713 TI - Molecular cloning and expression of a novel glycolipid sulfotransferase in Mycobacterium tuberculosis. AB - Sulfated trehalose glycolipids are among the most characteristic cell wall molecules of virulent strains of Mycobacterium tuberculosis. They comprise a family of trehalose-2-sulfate esters with an array of acyl fatty acids at various positions of the trehalose moiety. Although their structure has been well characterized, most of the enzymes involved in their biosynthesis, such as sulfotransferases, are unknown. It is demonstrated here by metabolic labelling with 35S abundant incorporation into sulfolipids of M. tuberculosis strains, in comparison to Mycobacterium avium, Mycobacterium bovis BCG and Mycobacterium smegmatis. The most abundant sulfolipid, sulfolipid I, is present in virulent strains H37Rv and Erdman, but absent in attenuated H37Ra. Sulfotransferase assays with the donor substrate 3'-phosphoadenosine-5'-[35S]phosphosulfonate and whole cell lysates of H37Ra resulted in the synthesis of four major sulfolipids (I, II, IV and VI). A search for sulfotransferase gene sequences in M. tuberculosis yielded gene Rv1373, a 981 bp gene slightly homologous (24% identity) to eukaryotic aryl-sulfotransferases. Rv1373 was cloned by PCR and expressed as a 39 kDa recombinant his-tagged protein. The recombinant M. tuberculosis aryl sulfotransferase exhibited activity towards the cerebroside glycolipids glucosyl- and galactosylceramide. No activity was detected with sulfatide (3'-sulfated galactosylceramide), suggesting that sulfation of galactosylceramide may occur at C-3 of the galactose. Treatment of sulfated products with ceramide glycanase resulted in the release of 35S-labelled material showing that sulfation was at the saccharide moiety (galactose or glucose) of the ceramide. Assays with the M. tuberculosis aryl-sulfotransferase and total H37Ra glycolipids showed one major product corresponding to sulfolipid IV. These results demonstrate that Rv1373 encodes a novel glycolipid sulfotransferase with activity towards typical ceramide glycolipids and mycobacterial trehalose glycolipids. PMID- 11882714 TI - Isocitrate lyase of the facultative intracellular pathogen Rhodococcus equi. AB - Isocitrate lyase is the first enzyme of the glyoxylate shunt which is required for the assimilation of fatty acids and acetate. The intracellular pathogen Rhodococcus equi contains high activities of this enzyme following growth on acetate and lactate, indicating that it plays an important role in the metabolism of these substrates. The gene encoding isocitrate lyase (aceA) was cloned and sequenced. It specifies a 46846 Da protein, which was shown to be functional by expressing it in Escherichia coli. A gene similar to fadB, encoding 3-hydroxyacyl CoA dehydrogenase, was located 90 bp downstream from aceA. Northern hybridization and RT-PCR experiments showed that aceA and fadB are cotranscribed into a 2.8 kb transcript. A smaller 1.6 kb aceA transcript was also observed which was 2.5-fold more abundant than the aceA-fadB transcript. It is proposed that a stable hairpin structure with a free energy (DeltaG) of -28.5 kcal x mol(-1) and located in the 90 bp aceA-fadB intergenic region is involved in stabilizing the aceA transcript. PMID- 11882715 TI - npd gene functions of Rhodococcus (opacus) erythropolis HL PM-1 in the initial steps of 2,4,6-trinitrophenol degradation. AB - Rhodococcus (opacus) erythropolis HL PM-1 grows on 2,4,6-trinitrophenol (picric acid) or 2,4-dinitrophenol (2,4-DNP) as sole nitrogen source. A gene cluster involved in picric acid degradation was recently identified. The functional assignment of three of its genes, npdC, npdG and npdI, and the tentative functional assignment of a fourth one, npdH, is reported. The genes were expressed in Escherichia coli as His-tag fusion proteins that were purified by Ni affinity chromatography. The enzyme activity of each protein was determined by spectrophotometry and HPLC analyses. NpdI, a hydride transferase, catalyses a hydride transfer from reduced F420 to the aromatic ring of picric acid, generating the hydride sigma-complex (hydride Meisenheimer complex) of picric acid (H(-)-PA). Similarly, NpdI also transformed 2,4-DNP to the hydride sigma complex of 2,4-DNP. A second hydride transferase, NpdC catalysed a subsequent hydride transfer to H(-)-PA, to produce a dihydride sigma-complex of picric acid (2H(-)-PA). All three reactions required the activity of NpdG, an NADPH-dependent F420 reductase, for shuttling the hydride ions from NADPH to F420. NpdH converted 2H(-)-PA to a hitherto unknown product, X. The results show that npdC, npdG and npdI play a key role in the initial steps of picric acid degradation, and that npdH may prove to be important in the later stages. PMID- 11882716 TI - Oligomerization of the Bacillus subtilis division protein DivIVA. AB - DivIVA appears to be a mediator of inhibition by MinCD of division at the cell poles in Bacillus subtilis. Gel permeation and ultracentrifugation techniques were used to show self-association of DivIVA into a form consisting of 10-12 monomers in vitro. Western blot analysis of non-denaturating polyacrylamide gels confirms the presence of similar oligomers in B. subtilis cell lysates. These oligomers persist in a B. subtilis strain containing the divIVA1 mutation, in which proper vegetative septum positioning is abolished. In contrast, the divIVA2 mutation, which has a similar biological impact, appears to produce a protein with different oligomerization properties. The results of the present study suggest that oligomerization of DivIVA is important, but not sufficient for its function in the cell division process. PMID- 11882717 TI - Comparison of ribitol and glycerol teichoic acid genes in Bacillus subtilis W23 and 168: identical function, similar divergent organization, but different regulation. AB - The tar genes directing the synthesis of poly(ribitol phosphate), the main teichoic acid in Bacillus subtilis strain W23, were sequenced. They are organized in two divergently transcribed operons, tarABIJKL and tarDF, as are the tag genes specifying poly(glycerol phosphate) synthesis in B. subtilis 168. The features of the tar genes as well as the putative participation of their products in the proposed biosynthesis pathway of poly(ribitol phosphate) are presented. The tarA and tarD genes, which are most likely involved in the synthesis of the linkage unit (the entity coupling teichoic acid to peptidoglycan), are separated by 508 nt. Sequences of the outer segments of this regulatory region are similar to the two divergent promoter regions identified upstream of the tagA and tagD genes in strain 168. However, in W23, these regions, which also included functional promoters, are separated by an additional DNA segment of about 100 nt, on which two new mRNA starts, one in each direction, were identified. The regulatory regions of teichoic acid divergons of Bacillus globigii, Bacillus licheniformis and eight strains of B. subtilis were cloned and sequenced. In four B. subtilis strains and in B. globigii, their length and sequence are similar to the regulatory region of W23. In the others, including B. licheniformis, they are of the 168-type. Analysis of nucleotide sequences of a non-coding grey hole, present in the tag region of strain 168, revealed higher similarities to tar than to tag entities. This suggests that at least part of the tag genes specifying the synthesis of glucosylated poly(glycerol phosphate) in strain 168 was introduced by horizontal gene transfer into a strain originally synthesizing a ribitol phosphate-containing teichoic acid. PMID- 11882718 TI - Characterization of a member of the NnrR regulon in Rhodobacter sphaeroides 2.4.3 encoding a haem-copper protein. AB - Upstream of the nor and nnrR cluster in Rhodobacter sphaeroides 2.4.3 is a previously uncharacterized gene that has been designated nnrS. nnrS is only expressed when 2.4.3 is grown under denitrifying conditions. Expression of nnrS is dependent on the transcriptional regulator NnrR, which also regulates expression of genes required for the reduction of nitrite to nitrous oxide, including nirK and nor. Deletion analysis indicated the sequence 5' TTGCG(N4)CACAA-3', which is similar to sequences found in nirK and nor, is required for nnrS expression. Mutation of this sequence to the consensus Fnr binding sequence by changing two bases in each half site caused nnrS expression to become nitrate independent. Inactivation of nnrS did not affect nitric oxide metabolism, nor did it affect expression of any of the genes involved in nitric oxide metabolism. However, taxis towards nitrate and nitrite was affected by nnrS inactivation. Purification of a histidine-tagged NnrS demonstrated that NnrS is a haem- and copper-containing membrane protein. Genes encoding putative orthologues of NnrS are sometimes but not always found in bacteria encoding nitrite and/or nitric oxide reductase. PMID- 11882719 TI - Regulation of the expression of prtW::gusA fusions in Erwinia carotovora subsp. carotovora. AB - Erwinia carotovora subsp. carotovora, a Gram-negative phytopathogenic bacterium, secretes an extracellular metalloprotease, PrtW. Previous results demonstrated that protease activity is necessary for the normal progression of disease symptoms caused by this bacterium. The present study revealed that the prtW gene constitutes an independent transcriptional unit. It is demonstrated that introduction of the prtW(+) plasmid in trans into the prtW(-) mutant restores the protease activity in this strain. Gene fusions to the gusA (beta-glucuronidase) reporter were employed to analyse the transcription of prtW. The transcription of prtW is dependent on many environmental signals. When the bacteria were grown in the presence of potato extract, the expression of the protease gene was markedly higher at the beginning of the exponential phase of growth than that observed when cells were grown in the presence of polygalacturonate (PGA). Analysis of the promoter revealed that an essential regulatory region resided between 371 and 245 bp 5' of the translational start site. As this sequence showed no homology to the KdgR box it may be involved in the binding of an unknown negative regulator protein in E. carotovora subsp. carotovora. The differential responses of prtW expression to potato extract and to PGA appeared to be dependent on the KdgR repressor and the response regulator ExpA. According to the results presented here, it is conceivable that the multiple regulatory network allows flexibility in the expression of the prtW gene during different stages of infection. PMID- 11882720 TI - Metabolism of sucrose and its five isomers by Fusobacterium mortiferum. AB - Fusobacterium mortiferum utilizes sucrose [glucose-fructose in alpha(1-->2) linkage] and its five isomeric alpha-D-glucosyl-D-fructoses as energy sources for growth. Sucrose-grown cells are induced for both sucrose-6-phosphate hydrolase (S6PH) and fructokinase (FK), but the two enzymes are not expressed above constitutive levels during growth on the isomeric compounds. Extracts of cells grown previously on the sucrose isomers trehalulose alpha(1-->1), turanose alpha(1-->3), maltulose alpha(1-->4), leucrose alpha(1-->5) and palatinose alpha(1-->6) contained high levels of an NAD+ plus metal-dependent phospho-alpha glucosidase (MalH). The latter enzyme was not induced during growth on sucrose. MalH catalysed the hydrolysis of the 6'-phosphorylated derivatives of the five isomers to yield glucose 6-phosphate and fructose, but sucrose 6-phosphate itself was not a substrate. Unexpectedly, MalH hydrolysed both alpha- and beta-linked stereomers of the chromogenic analogue p-nitrophenyl glucoside 6-phosphate. The gene malH is adjacent to malB and malR, which encode an EII(CB) component of the phosphoenolpyruvate-dependent sugar:phosphotransferase system and a putative regulatory protein, respectively. The authors suggest that for F. mortiferum, the products of malB and malH catalyse the phosphorylative translocation and intracellular hydrolysis of the five isomers of sucrose and of related alpha linked glucosides. Genes homologous to malB and malH are present in both Klebsiella pneumoniae and the enterohaemorrhagic strain Escherichia coli O157:H7. Both these organisms grew well on sucrose, but only K. pneumoniae exhibited growth on the isomeric compounds. PMID- 11882721 TI - Bioenergetics of the alkaliphilic sulfate-reducing bacterium Desulfonatronovibrio hydrogenovorans. AB - Energy metabolism of the alkaliphilic sulfate-reducing bacterium Desulfonatronovibrio hydrogenovorans strain Z-7935 was investigated in continuous culture and in physiological experiments on washed cells. When grown in chemostats with H2 as electron donor, the cells had extrapolated growth yields [Y(max), g dry cell mass (mol electron acceptor)(-1)] of 5.5 with sulfate and 12.8 with thiosulfate. The maintenance energy coefficients were 1.9 and 1.3 mmol (g dry mass)(-1) x h(-1), and the minimum doubling times were 27 and 20 h with sulfate and thiosulfate, respectively. Cell suspensions reduced sulfate, thiosulfate, sulfite, elemental sulfur and molecular oxygen in the presence of H2. In the absence of H2, sulfite, thiosulfate and sulfur were dismutated to sulfide and sulfate. Sulfate and sulfite were only reduced in the presence of sodium ions, whereas sulfur was reduced also in the absence of Na+. Plasmolysis experiments showed that sulfate entered the cells via an electroneutral symport with Na+ ions. The presence of an electrogenic Na+-H+ antiporter was demonstrated in experiments applying monensin (an artificial electroneutral Na+-H+ antiporter) and propylbenzylylcholine mustard.HCl (a specific inhibitor of Na+-H+ antiporters). Sulfate reduction was sensitive to uncouplers (protonophores), whereas sulfite reduction was not affected. Changes in pH upon lysis of washed cells with butanol indicated that the intracellular pH was lower than the optimum pH for growth (pH 9.5). Pulses of NaCl (0.52 M) to cells incubated in the absence of Na+ did not result in ATP formation, whereas HCl pulses (shifting the pH from 9.2 to 7.0) did. Small oxygen pulses, which were reduced within a few seconds, caused a transient alkalinization. The results of preliminary experiments with chemiosmotic inhibitors provided further evidence that the alkalinization was caused by sodium--proton antiport following a primary electron-transport-driven sodium ion translocation. It is concluded that energy conservation in D. hydrogenovorans depends on a proton-translocating ATPase, whereas electron transport appears to be coupled to sodium ion translocation. PMID- 11882722 TI - The NtcA-activated amt1 gene encodes a permease required for uptake of low concentrations of ammonium in the cyanobacterium Synechococcus sp. PCC 7942. AB - In the unicellular cyanobacterium Synechococcus sp. PCC 7942, ammonium/methylammonium transport activity has been characterized but ammonium transport genes have not been described. The amt1 gene encoding a permease responsible for high-affinity [14C]methylammonium transport in Synechococcus sp. PCC 7942 was cloned and inactivated. The Amt1 permease appeared essential to take up ammonium when it was present at low concentrations in the external medium and might also be involved in recapture of ammonium leaked out from the cells. Expression of amt1, which was induced in the absence of ammonium and also influenced by the inorganic carbon supply, was dependent on the NtcA transcriptional regulator. The promoter of amt1 was found to exhibit the structure of NtcA-activated promoters, and specific binding of purified NtcA to amt1 promoter sequences was observed. The results of this study indicate that amt1 belongs to the NtcA regulon and that NtcA may respond to both nitrogen and carbon availability. PMID- 11882723 TI - Characterization of the replicator region of megaplasmid pTAV3 of Paracoccus versutus and search for plasmid-encoded traits. AB - The replicon of the pTAV3 megaplasmid (approx. 400 kb) of Paracoccus versutus has been localized to a 4center dot3 kb EcoRI restriction fragment and its entire nucleotide sequence determined. The G+C content of the entire sequence is 66 mol%, which is within the range (62-66 mol%) previously determined for P. versutus total DNA. ORF1 encodes a replication initiation protein Rep (47.2 kDa), which shares substantial similarity with putative proteins of the Coxiella burnetii plasmids QpH1 and QpDV, and the replication protein of Pseudomonas syringae plasmid pPS10. ORF2, located in the opposite transcriptional orientation to ORF1, encodes a putative protein that shares similarity to a subfamily of ATPases involved in plasmid partitioning. The highest similarity was observed with homologous proteins (RepA) encoded by the repABC family of replicons found in several plasmids of Agrobacterium, Rhizobium and Paracoccus spp. The predicted product of ORF3 was similar to AcoR, Nif and NtrC transcriptional activators. A strong incompatibility determinant (inc) was localized between ORF1 (rep) and ORF2 (parA). The origin of replication of pTAV400 contains a short A+T-rich region and several imperfect palindromic sequences. Curing experiments demonstrated that the megaplasmid bears genes required for growth in minimal media and can therefore be referred to as a mini-chromosome. Megaplasmids pTAV3 of P. versutus UW1 and pKLW2 of Paracoccus pantotrophus DSM 11073 were found to carry closely related, incompatible replicons. It has been shown that plasmid pORI6 (containing oriV of pTAV3 cloned into plasmid pABW1, which does not replicate in Paracoccus spp.) can be trans activated not only by pTAV3, but also by pKLW2. Using pORI6, it was demonstrated that replication systems related to pTAV3 are also present in the replicons of Paracoccus alcaliphilus JCM 7364, Paracoccus thiocyanatus IAM 12816 and Paracoccus methylutens DM 12. PMID- 11882724 TI - Functional replacement of the Escherichia coli hfq gene by the homologue of Pseudomonas aeruginosa. AB - The 102 aa Hfq protein of Escherichia coli (Hfq(Ec)) was first described as a host factor required for phage Qbeta replication. More recently, Hfq was shown to affect the stability of several E. coli mRNAs, including ompA mRNA, where it interferes with ribosome binding, which in turn results in rapid degradation of the transcript. In contrast, Hfq is also required for efficient translation of the E. coli and Salmonella typhimurium rpoS gene, encoding the stationary sigma factor. In this study, the authors have isolated and characterized the Hfq homologue of Pseudomonas aeruginosa (Hfq(Pa)), which consists of only 82 aa. The 68 N-terminal amino acids of Hfq(Pa) show 92% identity with Hfq(Ec). Hfq(Pa) was shown to functionally replace Hfq(Ec) in terms of its requirement for phage Qbeta replication and for rpoS expression. In addition, Hfq(Pa) exerted the same negative effect on E. coli ompA mRNA expression. As judged by proteome analysis, the expression of either the plasmid-borne hfq(Pa) or the hfq(Ec) gene in an E. coli Hfq(-) RpoS(-) strain revealed no gross difference in the protein profile. Both Hfq(Ec) and Hfq(Pa) affected the synthesis of approximately 26 RpoS independent E. coli gene products. These studies showed that the functional domain of Hfq resides within its N-terminal domain. The observation that a C terminally truncated Hfq(Ec) lacking the last 27 aa [Hfq(Ec(75))] can also functionally replace the full-length E. coli protein lends further support to this notion. PMID- 11882725 TI - Resistant bacteria in retail meats and antimicrobial use in animals. PMID- 11882726 TI - Attacking the pneumococcus -- a hundred years' war. PMID- 11882727 TI - The risk of major birth defects after intracytoplasmic sperm injection and in vitro fertilization. AB - BACKGROUND: It is not known whether infants conceived with use of intracytoplasmic sperm injection or in vitro fertilization have a higher risk of birth defects than infants conceived naturally. METHODS: We obtained data from three registries in Western Australia on births, births after assisted conception, and major birth defects in infants born between 1993 and 1997. We assessed the prevalence of major birth defects diagnosed by one year of age in infants conceived naturally or with use of intracytoplasmic sperm injection or in vitro fertilization. RESULTS: Twenty-six of the 301 infants conceived with intracytoplasmic sperm injection (8.6 percent) and 75 of the 837 infants conceived with in vitro fertilization (9.0 percent) had a major birth defect diagnosed by one year of age, as compared with 168 of the 4000 naturally conceived infants (4.2 percent; P<0.001 for the comparison between either type of technology and natural conception). As compared with natural conception, the odds ratio for a major birth defect by one year of age, after adjustment for maternal age and parity, the sex of the infant, and correlation between siblings, was 2.0 (95 percent confidence interval, 1.3 to 3.2) with intracytoplasmic sperm injection, and 2.0 (95 percent confidence interval, 1.5 to 2.9) with in vitro fertilization. Infants conceived with use of assisted reproductive technology were more likely than naturally conceived infants to have multiple major defects and to have chromosomal and musculoskeletal defects. CONCLUSIONS: Infants conceived with use of intracytoplasmic sperm injection or in vitro fertilization have twice as high a risk of a major birth defect as naturally conceived infants. PMID- 11882728 TI - Low and very low birth weight in infants conceived with use of assisted reproductive technology. AB - BACKGROUND: The increased risk of low birth weight associated with the use of assisted reproductive technology has been attributed largely to the higher rate of multiple gestations associated with such technology. It is uncertain, however, whether singleton infants conceived with the use of assisted reproductive technology may also have a higher risk of low birth weight than those who are conceived spontaneously. METHODS: We used population-based data to compare the rates of low birth weight (less-than-or-equal 2500 g) and very low birth weight (<1500 g) among infants conceived with assisted reproductive technology with the rates in the general population. RESULTS: We studied 42,463 infants who were born in 1996 and 1997 and conceived with assisted reproductive technology and used as a comparison group 3,389,098 infants born in the United States in 1997. Among singleton infants born at 37 weeks of gestation or later, those conceived with assisted reproductive technology had a risk of low birth weight that was 2.6 times that in the general population (95 percent confidence interval, 2.4 to 2.7). The use of assisted reproductive technology was associated with an increased rate of multiple gestations; however, its use was not associated with a further increase in the risk of low birth weight in multiple births. Among twins, the ratio of the rate of low birth weight after the use of assisted reproductive technology to the rate in the general population was 1.0 (95 percent confidence interval, 1.0 to 1.1). Infants conceived with assisted reproductive technology accounted for 0.6 percent of all infants born to mothers who were 20 years of age or older in 1997, but for 3.5 percent of low-birth-weight and 4.3 percent of very low-birth-weight infants. CONCLUSIONS: The use of assisted reproductive technology accounts for a disproportionate number of low-birth-weight and very low-birth-weight infants in the United States, in part because of absolute increases in multiple gestations and in part because of higher rates of low birth weight among singleton infants conceived with this technology. PMID- 11882729 TI - Hepatocytes and epithelial cells of donor origin in recipients of peripheral blood stem cells. AB - BACKGROUND: Bone marrow contains stem cells with the potential to differentiate into mature cells of various organs. We determined whether circulating stem cells have a similar potential. METHODS: Biopsy specimens from the liver, gastrointestinal tract, and skin were obtained from 12 patients who had undergone transplantation of hematopoietic stem cells from peripheral blood (11 patients) or bone marrow (1 patient). Six female patients had received transplants from a male donor. Five had received a sex-matched transplant, and one had received an autologous transplant. Hematopoietic stem-cell engraftment was verified by cytogenetic analysis or restriction-fragment--length polymorphism analysis. The biopsies were studied for the presence of donor-derived epithelial cells or hepatocytes with the use of fluorescence in situ hybridization of interphase nuclei and immunohistochemical staining for cytokeratin, CD45 (leukocyte common antigen), and a hepatocyte-specific antigen. RESULTS: All six recipients of sex mismatched transplants showed evidence of complete hematopoietic donor chimerism. XY-positive epithelial cells or hepatocytes accounted for 0 to 7 percent of the cells in histologic sections of the biopsy specimens. These cells were detected in liver tissue as early as day 13 and in skin tissue as late as day 354 after the transplantation of peripheral-blood stem cells. The presence of donor cells in the biopsy specimens did not seem to depend on the intensity of tissue damage induced by graft-versus-host disease. CONCLUSIONS: Circulating stem cells can differentiate into mature hepatocytes and epithelial cells of the skin and gastrointestinal tract. PMID- 11882730 TI - Resistance to levofloxacin and failure of treatment of pneumococcal pneumonia. PMID- 11882731 TI - Images in clinical medicine. Massive air embolism in a neonate with pulmonary hypoplasia. PMID- 11882732 TI - The antiphospholipid syndrome. PMID- 11882733 TI - Clinical problem-solving. Where are you from? PMID- 11882734 TI - Infertility treatment--more risks and challenges. PMID- 11882735 TI - Can human hematopoietic stem cells become skin, gut, or liver cells? PMID- 11882736 TI - Ooplasmic transfer--proceed with care. PMID- 11882737 TI - Resistant bacteria in retail meats and antimicrobial use in animals. PMID- 11882738 TI - Transfusion in elderly patients with myocardial infarction. PMID- 11882739 TI - Infective endocarditis. PMID- 11882740 TI - Hypochondriasis. PMID- 11882741 TI - Aortic pseudocoarctation causing refractory hypertension. PMID- 11882742 TI - Case 27-2001: Waldenstrom's macroglobulinemia. PMID- 11882743 TI - Mitochondrial genetic variants and Alzheimer disease: a case-control study of the T4336C and G5460A variants. AB - The T4336C mitochondrial genetic variant was associated with Alzheimer disease in several previous studies. Recent investigations, however, failed to confirm this association. We tested this association in newly diagnosed Alzheimer disease cases and controls of similar age and gender recruited from an established HMO serving Seattle, Washington and surrounding areas. In this, the largest case control study reported to date, the T4336C variant was not associated with Alzheimer disease overall (present in 6 of 236 cases and 7 of 328 controls; odds ratio = 1.20, 95% CI 0.33 to 4.22). There was evidence of effect modification by Apolipoprotein E (APOE) status--among subjects with an APOE epsilon 4 allele, the T4336C variant was associated with disease (present in 5 of 139 cases and none of 82 controls; odds ratio = infinity, 95% CI 0.73 to infinity). APOE may be an important modifier of the T4336C effect, potentially explaining variable findings across previous studies. Alternatively, the positive findings reported to date may simply reflect the problem of "type I" error inherent in genetic association studies. Substantially larger samples than are currently available would be required to resolve this question. G5460(A/T) variants were also investigated and found not to be associated with Alzheimer disease. PMID- 11882744 TI - Apolipoprotein E polymorphism and cognitive impairment in a bi-ethnic community dwelling elderly sample. AB - The epsilon 4 (epsilon 4) and epsilon 2 (epsilon 2) alleles of the apolipoprotein gene (APOE) located on chromosome 19 have been associated with increased and decreased risk for Alzheimer disease (AD) in older adults, respectively. However, there is a dearth of studies examining the relation of APOE polymorphism with cognitive functioning among community-dwelling ethnic minority elderly. This study examined the risk for cognitive impairment associated with the APOE epsilon 4 and epsilon 2 alleles in a community-based cohort of non-Hispanic white (NHW; N = 739) and white Hispanics (WH; N=321). All patients were recruited consecutively from a memory-screening program and evaluated using standardized assessment procedures. Cognitive impairment was classified according to an age and education adjusted Folstein Mini-Mental State Exam (MMSAdj) score of less than 24. The results indicated the APOE epsilon 4 allele was associated with increased risk for cognitive dysfunction in NHW and WH after controlling for the effects of age, education, and gender. This risk was generally observed to be dose-dependent, with greater risk among epsilon 4 homozygotes in relation to epsilon 4 heterozygotes. The epsilon 2 allele of APOE did not confer decreased risk for cognitive impairment among NHW and WH. This study supports the relation of APOE polymorphism to cognitive dysfunction among two ethnic populations residing in the community. PMID- 11882746 TI - H2 histamine receptor blockade in the treatment of Alzheimer disease: a randomized, double-blind, placebo-controlled trial of nizatidine. AB - OBJECTIVES: To evaluate the efficacy of nizatidine, a histamine H2-blocking drug, in delaying the progression of cognitive impairment in older adults with Alzheimer disease (AD). DESIGN: A one-year, randomized, double-blind, placebo controlled trial. PARTICIPANTS: Fifty-one older men and women aged 67 to 96 years with AD were recruited from the Cache County Study on Memory in Aging. METHODS: Patients were stratified by age and by the presence of one or more epsilon 4 alleles at the APOE locus, then randomized to receive nizatidine 75 mg (Axid ARTM, Whitehall Robins) or a matching placebo tablet twice daily. Cognitive outcomes were assessed at baseline, six, and twelve months after enrollment using tests from the CERAD battery and additional measures of visuospatial memory, verbal memory, and verbal fluency. RESULTS: Subjects showed significant declines in language, fluency, and praxis but most measures of memory had already "bottomed out." Intention-to-treat and compliance-based analyses showed no effect of nizatidine on any of the cognitive outcome measures over the one-year study interval. CONCLUSIONS: These results do not support claims for the efficacy of nizatidine in over-the-counter dosages as a means of preventing symptom progression in AD. PMID- 11882745 TI - How well are clinicians following dementia practice guidelines? AB - Although there are numerous clinical guidelines regarding the management of dementia, there have been few studies on their implementation in practice. Clinicians in six United States Department of Veterans Affairs medical centers (n = 200, 85% response rate) were surveyed regarding their use of practices recommended in the California Workgroup Guidelines for Alzheimer's Disease Management. The majority of providers (89% to 73%) reported that they routinely conducted neurological examinations, obtained histories from caregivers, discussed the diagnosis with the patient's family, discussed durable power of attorney, and made legally-required reports of drivers with dementia. Roughly two thirds of providers said they routinely conducted cognitive screening examinations, screened for depression, reported elder abuse, and discussed care needs and decision-making issues with patients' families. Only half of all outpatient providers implemented caregiver support practices for at least half of their patients. Clinicians' choices of medications for cognition, mood, and behavior problems were broadly consistent with current practice guidelines. These results suggest possible priorities for quality improvement efforts. Further research is needed to clarify reasons for particular gaps between guidelines and practice and to identify specific targets for intervention. PMID- 11882747 TI - Word-reading thresholds in Alzheimer disease and mild memory loss: a pilot study. AB - PURPOSE: To determine whether subjects with early Alzheimer Disease (AD) or Mild Cognitive Impairment (MCI) would demonstrate significant abnormalities on tests of word reading threshold (WRT), spatial contrast sensitivity (SCS), and color discrimination (CD). METHODS: Prospective cohort study of 13 AD subjects, 13 subjects with Mild Cognitive Impairment (MCI), and 12 healthy elderly normal controls (ENC). CD was determined using the Farnsworth D-15 method. SCS was tested using a Vistech photographic chart. For WRT determination, pattern-masked words were presented successively at increasing target durations. The threshold was determined as the target duration at which 50% of the words were read. RESULTS: The mean number of errors in Color Discrimination and Spatial Contrast Sensitivity did not differ significantly among the three groups. WRT was significantly longer in the AD group (122.6 +/- 70.8 ms) in comparison with ENC group (53.8 +/- 15.9 ms) (p < 0.001). A WRT threshold limit of 85 msec correctly classified 11/12 ENC subjects and 10/13 AD subjects. Four of the 13 MCI subjects also exceeded this threshold and all four progressed to AD within 2 year follow up. CONCLUSION: Word Reading Threshold determination discriminates mild AD subjects from ENC, and correlates with severity of cognitive impairment. It may have diagnostic and prognostic utility in early AD and MCI subjects. PMID- 11882748 TI - Theta-power differences in patients with mild cognitive impairment under rest condition and during haptic tasks. AB - The aim of this study was to investigate spectral EEG theta-power during perceptive-cognitive demands in age-homogeneous groups of subjects with mild cognitive impairment (MCI), mild dementia (MDE), and a healthy control (CO) group. The present study includes 51 subjects (23 males, 28 females). We used the scales of the CDR (clinical dementia rating) to assign the subjects to the different groups. EEG data were collected during 10 minutes rest condition with eyes closed and during haptic perception test. The quality of the haptic reproductions differed significantly between CO and MCI, as well as between CO and MDE. The statistical comparison between EEG theta-power under rest condition and theta-power during haptic tasks revealed a significant decrease in theta power during haptic tasks in all three groups over parieto-occipital regions. During haptic tasks, the theta-power was significantly different between CO and MDE over occipital regions and over parieto-temporal regions. A significant difference between CO and MCI was only revealed over right occipital regions (O2). Spectral theta-power during haptic tasks is a suitable measure to distinguish healthy subjects (CO) from patients with MCI respectively MDE. The results show that haptic tasks are sensitive to early perceptive-cognitive and functional deficits in patients with MCI. PMID- 11882749 TI - How many patients complete an adequate trial of donepezil? AB - Pharmacy claims data were used to evaluate medication adherence among 59 new users of donepezil aged 65 to 94 years. The probability (+/- 95% confidence interval) of a new user continuing donepezil at 90 days was.797 +/-.103 and at 180 days was.627 +/-.124. Additionally, 13.9% of those who continued therapy for at least 180 days showed gaps in treatment of six weeks or more. These results suggest that adherence with donepezil could be improved in clinical practice. PMID- 11882750 TI - Blood platelet membrane fluidity and the exposition of membrane protein receptors in Alzheimer disease (AD) patients--preliminary Study. AB - We estimated membrane fluidity by the (ESR) spectroscopy and the expression of membrane P-selectin and glycoproteins GP Ib alpha and GP IIb/IIIa by flow cytometry in platelets (plts) from 12 AD sufferers. In AD patients membrane fluidity was significantly increased at two different depths (p < 0.05 or less). Platelet reactivity was significantly decreased, as reflected by reduced expression of GP Ib alpha in the resting (p < 0.0001) and activated (p < 0.005) platelets, as well as the expression of P-selectin and beta 3 subunit of GP IIb/IIIa in activated platelets (p < 0.0001; p < 0.04). PMID- 11882752 TI - Laparoscopic repair for perforated peptic ulcer. PMID- 11882751 TI - Laparoscopic repair for perforated peptic ulcer: a randomized controlled trial. AB - OBJECTIVE: To compare the results of open versus laparoscopic repair for perforated peptic ulcers. SUMMARY BACKGROUND DATA: Omental patch repair with peritoneal lavage is the mainstay of treatment for perforated peptic ulcers in many institutions. Laparoscopic repair has been used to treat perforated peptic ulcers since 1990, but few randomized studies have been carried out to compare open versus laparoscopic procedures. METHODS: From January 1994 to June 1997, 130 patients with a clinical diagnosis of perforated peptic ulcer were randomly assigned to undergo either open or laparoscopic omental patch repair. Patients were excluded for a history of upper abdominal surgery, concomitant evidence of bleeding from the ulcer, or gastric outlet obstruction. Patients with clinically sealed-off perforations without signs of peritonitis or sepsis were treated without surgery. Laparoscopic repair would be converted to an open procedure for technical difficulties, nonjuxtapyloric gastric ulcers, or perforations larger than 10 mm. A Gastrografin meal was performed 48 to 72 hours after surgery to document sealing of the perforation. The primary end-point was perioperative parenteral analgesic requirement. Secondary endpoints were operative time, postoperative pain score, length of postoperative hospital stay, complications and deaths, and the date of return to normal daily activities. RESULTS: Nine patients with a surgical diagnosis other than perforated peptic ulcer were excluded; 121 patients entered the final analysis. There were 98 male and 23 female patients recruited, ages 16 to 89 years. The two groups were comparable in age, sex, site and size of perforations, and American Society of Anesthesiology classification. There were nine conversions in the laparoscopic group. After surgery, patients in the laparoscopic group required significantly less parenteral analgesics than those who underwent open repair, and the visual analog pain scores in days 1 and 3 after surgery were significantly lower in the laparoscopic group as well. Laparoscopic repair required significantly less time to complete than open repair. The median postoperative stay was 6 days in the laparoscopic group versus 7 days in the open group. There were fewer chest infections in the laparoscopic group. There were two intraabdominal collections in the laparoscopic group. One patient in the laparoscopic group and three patients in the open group died after surgery. CONCLUSIONS: Laparoscopic repair of perforated peptic ulcer is a safe and reliable procedure. It was associated with a shorter operating time, less postoperative pain, reduced chest complications, a shorter postoperative hospital stay, and earlier return to normal daily activities than the conventional open repair. PMID- 11882753 TI - Repair of groin hernia with synthetic mesh: meta-analysis of randomized controlled trials. AB - OBJECTIVE: To measure the effects of laparoscopic and open placement of synthetic mesh on recurrence and persisting pain following groin hernia repair. SUMMARY BACKGROUND DATA: Synthetic mesh techniques are claimed to reduce the risk of recurrence but there are concerns about costs and possible long-term complications, particularly pain. METHODS: Electronic databases were searched and experts consulted to identify randomized or quasi-randomized trials that compared mesh with non-mesh methods, or laparoscopic with open mesh placement. Individual patient data were sought for each trial. Aggregated data were used where individual patient data were not available. Meta-analyses of hernia recurrence and persisting pain were based on intention to treat. RESULTS: There were 62 relevant comparisons in 58 trials. These included 11,174 participants: individual patient data were available for 6,901 patients, supplementary aggregated data for 2,390 patients, and published data for 1883 patients. Recurrence and persisting pain were less after mesh repair (overall recurrences: 88 in 4,426 vs. 187 in 3,795; OR 0.43, 95% CI 0.34-0.55; P <.001) (overall persistent pain: 120 in 2,368 vs. 215 in 1,998; OR 0.36, 95% CI 0.29-0.46; P <.001), regardless of the non-mesh comparator. Whereas the reduction in recurrence was similar after laparoscopic and open mesh placement (OR 1.26, 95% CI 0.76-2.08; P =.36), persistent pain was less common after laparoscopic than open mesh placement (OR 0.64; 95% CI 0.52 0.78; P <.001). CONCLUSIONS: The use of synthetic mesh substantially reduces the risk of hernia recurrence irrespective of placement method. Mesh repair appears to reduce the chance of persisting pain rather than increase it. PMID- 11882754 TI - Five-year follow-up of patients undergoing laparoscopic or open groin hernia repair: a randomized controlled trial. AB - OBJECTIVE: To compare laparoscopic with open hernia repair in a randomized clinical trial at a median follow-up of 5 years. SUMMARY BACKGROUND DATA: Follow up of patients in clinical trials evaluating laparoscopic hernia repair has been short. METHODS: Of 379 consecutive patients admitted for surgery under the care of one surgeon, 300 were randomized to totally extraperitoneal hernia repair or open repair, with the open operation individualized to the patient's age and hernia type. All patients, both randomized and nonrandomized, were followed up by clinical examination annually by an independent observer. RESULTS: Recurrence rates were similar for both randomized groups. In 1 of the 79 nonrandomized patients, a recurrent hernia developed. Groin or testicular pain was the most common symptom on follow-up of randomized patients. The most common reason for reoperation was development of a contralateral hernia, which was noted in 9% of patients; 11% of all patients died on follow-up, mainly as a result of cardiovascular disease or cancer. CONCLUSIONS: These data show a similar outcome for laparoscopic and open hernia repair, and both procedures have a place in managing this common problem. PMID- 11882755 TI - Metaplastic columnar mucosa in the cervical esophagus after esophagectomy. AB - OBJECTIVE: To evaluate the pathogenesis of metaplastic processes within the esophagus using a human model in which the exact duration of reflux was known. SUMMARY BACKGROUND DATA: The pathogenesis of Barrett's esophagus (BE) is incompletely understood. Patients undergoing esophagectomy and gastric tube reconstruction represent a good model for studying the pathophysiology of columnar cell metaplasia of the human esophagus because the cervical esophagus is rarely or never exposed to gastric contents before the surgical procedure. METHODS: Thirty-two patients underwent manometry, simultaneous 24-hour pH and bilirubin monitoring, and endoscopy with biopsy 3 to 10.4 years after esophagectomy. The presence of columnar mucosa in the cervical esophagus was confirmed on histologic examination. The findings on endoscopy and histology were related to clinical data and the results of pH and bilirubin monitoring 1 cm proximal to the esophagogastrostomy. RESULTS: Fifteen (46.9%) of the 32 patients had metaplastic columnar mucosa within their cervical esophagus. Metaplasia was significantly more common in patients with a preoperative diagnosis of BE. The length of metaplastic mucosa correlated significantly with the degree of esophageal acid exposure, but the presence of abnormal bilirubin exposure was unrelated to the presence of metaplasia. The prevalence of metaplasia did not change with increasing time. Intestinal metaplasia was found within the columnar lined segment in three patients 8.5, 9.5, and 10.4 years after esophagectomy. All patients with intestinal metaplasia had abnormal exposure of both acid and bilirubin, but the presence of combined reflux was not significantly higher in these patients compared with patients with nonintestinalized segments of columnar mucosa. CONCLUSIONS: Esophageal columnar metaplasia is a common complication after gastric pull-up esophagectomy. Metaplasia is more likely to develop in patients with previous BE than other patients. Metaplasia develops in response to squamous epithelial injury in predisposed individuals. PMID- 11882756 TI - Physiologic basis for the treatment of epiphrenic diverticulum. AB - OBJECTIVE: To quantitate and characterize the motility abnormalities present in patients with epiphrenic diverticula and to assess the outcome of surgical treatment undertaken according to these abnormalities. SUMMARY BACKGROUND DATA: The concept that epiphrenic diverticula are complications of esophageal motility disorders rather than primary anatomic abnormalities is gradually becoming accepted. The inconsistency in identifying motility abnormalities in patients with epiphrenic diverticula is a major obstacle to the general acceptance of this concept. METHODS: The study population consisted of 21 consecutive patients with epiphrenic diverticula. All patients underwent videoesophagography, upper gastrointestinal endoscopy, and esophageal motility studies. The diverticula ranged in size from 3 to 10 cm and were predominantly right-sided. Seventeen patients underwent transthoracic diverticulectomy or diverticulopexy with esophageal myotomy and an antireflux procedure. The length of the myotomy was determined by the extent of the motility abnormality. Transhiatal esophagectomy was performed in one patient with multiple diverticula. Two patients declined surgical treatment and another patient died of aspiration before surgery. Symptomatic outcome was assessed via a questionnaire at a median of 24 months after surgery. RESULTS: The primary symptoms were dysphagia in 5 (24%) patients, dysphagia and regurgitation in 11 (52%) patients, and pulmonary symptoms in 5 (24%) patients. The median duration of the primary symptoms was 10 years. Esophageal motility abnormalities were identified in all patients. An esophageal motor disorder was diagnosed only by 24-hour ambulatory motility testing in one patient, and 24-hour ambulatory motility testing clarified the motility diagnosis in five other patients. The most common underlying disorder was achalasia, which was detected in nine (43%) patients. A hypertensive lower esophageal sphincter was diagnosed in three patients, diffuse esophageal spasm in five, "nutcracker" esophagus in two, and a nonspecific motor disorder in two patients. One patient had an intraoperative myocardial infarction and died. Two patients had persistent mild dysphagia after surgery. The remaining patients had complete relief of their primary symptoms. CONCLUSIONS: There is a high prevalence of named motility disorders in patients with epiphrenic diverticula, and this condition is associated with the potential for lethal aspiration. Twenty-four-hour ambulatory motility testing can be helpful if the results of the stationary examination are normal or indefinite. Resection of the diverticula and a surgical myotomy of the manometrically defined abnormal segment results in relief of symptoms and protection from aspiration. PMID- 11882757 TI - Ten-year experience of endoscopic transanal resection. AB - OBJECTIVE: To audit the results of endoscopic transanal resection of tumor (ETAR) performed by a single surgeon at a specialized colorectal unit during a 10- year period. SUMMARY BACKGROUND DATA: A minimally invasive surgical technique, ETAR has enabled much progress to be made in the development of local treatment strategies for rectal neoplasia. It can be used in both the curative and palliative management of rectal lesions and is a treatment option for patients who would be unable to tolerate major surgery. METHODS: The surgical outcome of 104 patients (43 women, 61 men) undergoing ETAR under the care of a single surgeon between 1989 and 1999 was reviewed. Patients were identified from the consultant's personal records and cross-referenced with operating room logs. Data were collected retrospectively and no patients were lost to follow-up. RESULTS: One hundred four patients underwent 163 procedures during the study period. Follow-up ranged from 6 months to 10 years. Seventy-five patients with a pre-ETAR diagnosis of benign rectal adenoma underwent resection. In 60 patients, the diagnosis was confirmed to be benign; 30 of these were treated with a single resection and 28 with multiple resections. There were two technical failures, both a result of high mobility of the lesion. In no patients did carcinoma subsequently develop. In the remaining 15 patients the final histology demonstrated a malignancy; 9 patients underwent an open surgical rectal resection and 5 had complete endoscopic resection of their lesion. No carcinomas that were fully resected endoscopically have recurred (follow-up 13 months to 8years). The final patient had an extensive rectal cancer and was palliated for 2 months by ETAR. Twelve patients (8 men, 4 women) underwent ETAR for anastomotic strictures; successful treatment was achieved in 11. The one failure was in a Park's pouch that was subsequently refashioned. Seventeen patients underwent 30 ETARs for palliation of nonresectable rectal adenocarcinoma. Successful palliation of symptoms was achieved in 13 patients and the remainder underwent colostomy formation. One patient died of a myocardial infarction. There were two further complications (blood transfusion for postoperative bleeding, postoperative cerebrovascular accident). CONCLUSIONS: Endoscopic transanal resection of tumor is safe and effective and offers successful palliation or definitive treatment of rectal lesions with low rates of death and complications when performed by a dedicated surgeon. PMID- 11882758 TI - Fulminant Clostridium difficile: an underappreciated and increasing cause of death and complications. AB - OBJECTIVE: To review the epidemiology and characteristics of patients who died or underwent colectomy secondary to fulminant Clostridium difficile colitis. SUMMARY BACKGROUND DATA: In patients with C. difficile colitis, a progressive, systemic inflammatory state may develop that is unresponsive to medical therapy; it may progress to colectomy or death. METHODS: The authors reviewed 2,334 hospitalized patients with C. difficile colitis from January 1989 to December 2000. Sixty-four patients died or underwent colectomy for pathologically proven C. difficile colitis. RESULTS: In 2000, the incidence of C. difficile colitis in hospitalized patients increased from a baseline of 0.68% to 1.2%, and the incidence of patients with C. difficile colitis in whom life-threatening symptoms developed increased from 1.6% to 3.2%. Forty-four patients required a colectomy and 20 others died directly from C. difficile colitis. Twenty-two percent had a prior history of C. difficile colitis. A recent surgical procedure and immunosuppression were common predisposing conditions. Lung transplant patients were 46 times more likely to have C. difficile colitis and eight times more likely to have severe disease. Abdominal computed tomography scan correctly diagnosed all patients, whereas 12.5% of toxin assays and 10% of endoscopies were falsely negative. Patients undergoing colectomy for C. difficile colitis had an overall death rate of 57%. Significant predictors of death after colectomy were preoperative vasopressor requirements and age. CONCLUSIONS: C. difficile colitis is a significant and increasing cause of death. Surgical treatment of C. difficile colitis has a high death rate once the fulminant expression of the disease is present. PMID- 11882759 TI - Long-term survival and pattern of recurrence after resection of small hepatocellular carcinoma in patients with preserved liver function: implications for a strategy of salvage transplantation. AB - OBJECTIVE: To evaluate the survival results and pattern of recurrence after resection of potentially transplantable small hepatocellular carcinomas (HCC) in patients with preserved liver function, with special reference to the implications for a strategy of salvage transplantation. SUMMARY BACKGROUND DATA: Primary resection followed by transplantation for recurrence or deterioration of liver function has been recently suggested as a rational strategy for patients with HCC 5 cm or smaller and preserved liver function. However, there are no published data on transplantability after HCC recurrence or long-term deterioration of liver function after resection of small HCC in Child-Pugh class A patients. Such data are critical in determining the feasibility of salvage transplantation. METHODS: From a prospective database of 473 patients with resection of HCC between 1989 and 1999, 135 patients age 65 years or younger had Child-Pugh class A chronic liver disease (chronic hepatitis or cirrhosis) and transplantable small HCC (solitary < or =5 cm or two or three tumors < or = 3 cm). Survival results were analyzed and the pattern of recurrence was examined for eligibility for salvage transplantation based on the same criteria as those of primary transplantation for HCC. RESULTS: Overall survival rates at 1, 3, 5, and 10 years were 90%, 76%, 70%, and 35%, respectively, and the corresponding disease-free survival rates were 74%, 50%, 36%, and 22%. Cirrhosis and oligonodular tumors were predictive of worse disease-free survival. Patients with concomitant oligonodular tumors and cirrhosis had a 5-year overall survival rate of 48% and a disease-free survival rate of 0%, which were significantly worse compared with other subgroups. At a median follow-up of 48 months, 67 patients had recurrence and 53 (79%) of them were considered eligible for salvage transplantation. Decompensation from Child-Pugh class A to B or C without recurrence occurred in only six patients. CONCLUSIONS: For Child-Pugh class A patients with small HCC, hepatic resection is a reasonable first-line treatment associated with a favorable 5-year overall survival rate. A considerable proportion of patients may survive without recurrence for 5 or even 10 years; among those with recurrence, the majority may be eligible for salvage transplantation. These data suggest that primary resection and salvage transplantation may be a feasible and rational strategy for patients with small HCC and preserved liver function. Primary transplantation may be a preferable option for the subset of patients with oligonodular tumors in cirrhotic liver in view of the poor survival results after resection. PMID- 11882760 TI - Hepatic abscess in patients with chronic granulomatous disease. AB - OBJECTIVE: To evaluate the clinical presentation, diagnostic procedures, and surgical management of hepatic abscesses in patients with chronic granulomatous disease (CGD). SUMMARY BACKGROUND DATA: Chronic granulomatous disease is a rare inherited primary immunodeficiency in which phagocytes cannot destroy catalase positive bacteria and fungi. Defects in the phagocytic cells' respiratory burst lead to life-threatening infections, including hepatic abscess. These abscesses are recurrent and often multiple and are treated differently from bacterial abscesses in patients without CGD. METHODS: Between 1980 and 2000, 61 cases of hepatic abscess in 22 patients with CGD were treated at the National Institutes of Health. Clinicopathologic features were investigated by retrospective review of the medical records, radiographs, and histopathology. RESULTS: Twelve of the 61 cases were primary hepatic abscesses. Twenty-nine of the cases were recurrent hepatic abscesses, and 20 cases were persistent hepatic abscesses. The median age at the time of initial hepatic abscess presentation was 14 years. Subjective fever was the most frequent presenting symptom, and the erythrocyte sedimentation rate was elevated in 98% of cases. Fifty-two cases were managed surgically and eight cases were managed with percutaneous drainage. One patient refused surgery. The surgical complication rate was 56%; however, there were no deaths directly related to the hepatic abscesses. Staphylococcus aureus was the most frequent organism identified in culture (88% of positive cultures). Aggressive surgery and antibiotics ultimately resulted in successful treatment of all patients. CONCLUSIONS: Hepatic abscesses occurring in patients with CGD represent a difficult diagnostic and treatment challenge. Early excision and treatment with antibiotics directed against S. aureus is necessary. General surgeons should be aware of this rare immunodeficiency and should aggressively manage hepatic abscesses in these patients. PMID- 11882761 TI - Staging laparoscopy in patients with extrahepatic biliary carcinoma. Analysis of 100 patients. AB - OBJECTIVE: To evaluate the benefit of staging laparoscopy in patients with gallbladder cancer and hilar cholangiocarcinoma. SUMMARY BACKGROUND DATA: In patients with extrahepatic biliary carcinoma, unresectable disease is often found at the time of exploration despite extensive preoperative evaluation, thus resulting in unnecessary laparotomy. METHODS: From October 1997 to May 2001, 100 patients with potentially resectable gallbladder cancer (n = 44) and hilar cholangiocarcinoma (n = 56) were prospectively evaluated. All patients underwent staging laparoscopy followed by laparotomy if the tumor appeared resectable. Surgical findings, resectability rate, length of stay, and operative time were analyzed. RESULTS: Patients underwent multiple preoperative imaging tests, including computed tomography scan, ultrasound, magnetic resonance cholangiopancreatography, and direct cholangiography. Laparoscopy identified unresectable disease in 35 of 100 patients. In the 65 patients undergoing open exploration, 34 were found to have unresectable disease. Therefore, the overall accuracy for detecting unresectable disease was 51%. There was no difference in the accuracy of laparoscopy between patients with gallbladder cancer and hilar cholangiocarcinoma. Laparoscopy detected the majority of patients with peritoneal or liver metastases but failed to detect all locally advanced tumors. In patients undergoing biopsy only, laparoscopic identification of unresectable disease significantly reduced operative time and length of stay compared with patients undergoing laparotomy. The yield of laparoscopy was 48% in patients with gallbladder cancer (56% in those who did not undergo previous cholecystectomy), but only 25% in patients with hilar cholangiocarcinoma. However, in patients with locally advanced but potentially resectable hilar cholangiocarcinoma, the yield of laparoscopy was greater, 36% (12/33, T2/T3 tumors) versus 9% (2/23, T1 tumors). CONCLUSIONS: Laparoscopy identifies the majority of patients with unresectable hilar cholangiocarcinoma or gallbladder carcinoma, thereby reducing both the incidence of unnecessary laparotomy and the length of stay. The yield of laparoscopy is lower for hilar cholangiocarcinoma but can be improved by targeting patients at higher risk of occult unresectable disease. All patients with potentially resectable primary gallbladder cancer and patients with T2/T3 hilar cholangiocarcinoma should undergo staging laparoscopy before surgical exploration. PMID- 11882762 TI - Comparison of ischemic preconditioning and intermittent and continuous inflow occlusion in the murine liver. AB - OBJECTIVE: To compare protection of the liver by ischemic preconditioning and intermittent inflow occlusion in a mouse model of prolonged periods of ischemia. SUMMARY BACKGROUND DATA: Preconditioning (short ischemic stress prior to a prolonged period of ischemia) and intermittent inflow occlusion protect the liver against reperfusion injury. This is the first study comparing these two modalities with continuous inflow occlusion (control). METHODS: Mice were subjected to 75 or 120 minutes of 70% hepatic ischemia and 3 hours of reperfusion. Each ischemic period was evaluated using three different protocols: continuous ischemia (control), preconditioning (10 minutes ischemia and 15 minutes reperfusion) prior to the prolonged ischemic insult, and intermittent clamping (cycles of 15 minutes ischemia and 5 minutes reperfusion). Organ injury was evaluated using serum levels of aspartate aminotransferase (AST), hematoxylin and eosin staining, and specific markers of apoptosis (cytochrome C release, caspase 3 activity, and TUNEL staining). Animal survival was determined using a model of total hepatic ischemia. RESULTS: Intermittent inflow occlusion and ischemic preconditioning were both protective against ischemic insults of 75 and 120 minutes compared with controls (continuous ischemia only). Protection against 75 minutes of ischemia was comparable in the intermittent clamping and the ischemic preconditioning group, whereas intermittent clamping was superior at 120 minutes of ischemia. One hundred percent animal survival was observed after 75 minutes of total hepatic ischemia using both protective protocols, whereas all animals subjected to continuous ischemia died after surgery. After 120 minutes of ischemia, intermittent inflow occlusion was associated with better animal survival (71%) compared with preconditioning (14%). CONCLUSIONS: Preconditioning and intermittent clamping are both protective against prolonged periods of ischemia. In the clinical setting, preconditioning is superior for ischemic periods of up to 75 minutes because it is not associated with blood loss during transection of the liver. However, for prolonged ischemic insults exceeding 75 minutes, intermittent clamping is superior to preconditioning. PMID- 11882763 TI - Metabolic control patterns in acute phase and regenerating human liver determined in vivo by 31-phosphorus magnetic resonance spectroscopy. AB - OBJECTIVE: To elucidate the metabolic changes occurring within hepatocytes during acute phase reaction and liver regeneration. SUMMARY BACKGROUND DATA: The metabolic events occurring within the liver during the hepatic stress response are poorly understood. The authors used in vivo 31-phosphorus magnetic resonance spectroscopy to study hepatic metabolism after surgical trauma with and without loss of liver cell mass. METHODS: Three groups were studied: five patients undergoing partial hepatectomy; five patients in whom laparotomy and colonic resection was performed; and five patients treated by thyroidectomy. Hepatic metabolism was evaluated by 31-phosphorus magnetic resonance spectroscopy before surgery and serially thereafter on postoperative days 2, 4, 6, 14, and 28. Estimation of liver volume by magnetic resonance imaging and blood sampling for biochemistry were performed at the same time points. RESULTS: The authors found that alterations in hepatocyte phospholipid metabolism occurred after surgery that were correlated with changes in circulating acute phase proteins. Liver regeneration after hepatectomy was also associated with a derangement in energy metabolism, measured by a decrease in the ratio of ATP to its hydrolysis product inorganic phosphate. The depleted energy status was mirrored in biochemical indices of liver function, and restitution paralleled the course of restoration of hepatic cell mass. CONCLUSIONS: These findings indicate that changes in liver metabolism after surgery reflect the magnitude of tissue injury and the quantity of functioning liver cells. Acute phase responses dominate the initial recovery period at the expense of less important endergonic functions. When liver parenchyma is lost, the acute phase reaction is maintained and further supported by a rapid replenishment of hepatocytes, which can even be considered a continuation of acute phase physiology. Modulation of liver function within the framework of overall hepatic energy economy is one mechanism for matching energy supply with increased demands during these processes. PMID- 11882764 TI - Gastric phase 3 motility after pylorus-preserving pancreatoduodenectomy. AB - OBJECTIVE: To analyze factors affecting the recovery course of phase 3 activity of the gastric migrating motor complex after pylorus-preserving pancreatoduodenectomy (PPPD) and investigate effects of the recovery of gastric phase 3 on gastric emptying after feeding. SUMMARY BACKGROUND DATA: Whether early recovery of gastric phase 3 during fasting would predict early recovery of the fed-state gastric emptying function after PPPD has not been well documented. METHODS: Manometric recording from the gastric antrum was repeated at a weekly interval until the first appearance of gastric phase 3 in 57 patients after PPPD. Twenty-three clinical parameters were assessed as possible factors affecting the recovery course of gastric phase 3 by simple and multiple regression analyses. A gastric emptying study after feeding of a test meal was performed by the acetaminophen method and the values were compared between patients with and without gastric phase 3 after PPPD. RESULTS: The mean period before the first appearance of gastric phase 3 was 38 days. Among 23 parameters, only lymph node dissection along the hepatoduodenal ligament significantly delayed recovery of gastric phase 3 after PPPD by univariate and multivariate analyses. The presence or absence of gastric phase 3 in the early postoperative period did not influence gastric emptying after feeding in the intermediate period after PPPD. CONCLUSIONS: Avoiding lymph node dissection along the hepatoduodenal ligament, if applicable, may contribute to early recovery of gastric phase 3 after PPPD. The recovery state of gastric phase 3 during fasting, however, is not necessarily consistent with the degree of improvement of gastric emptying after feeding. PMID- 11882765 TI - Analysis of the prognostic significance of microscopic margins in 2,084 localized primary adult soft tissue sarcomas. AB - OBJECTIVE: To define the significance of positive microscopic resection margins in a large cohort treated for soft tissue sarcoma. METHODS: The authors analyzed 2,084 patients with localized primary soft tissue sarcoma (all anatomic sites) treated from 1982 to 2000. Clinicopathologic variables studied included tumor site, size, depth, histologic type, grade, and resection margin status. Treatment other than resection was not analyzed. Study endpoints included local and distant recurrence-free and disease-specific survival rates, estimated by the Kaplan Meier method. Univariate and multivariate analyses were performed using the log rank test and the Cox proportional hazards model. RESULTS: Median follow-up was 50 months. After primary resection, 1,624 (78%) patients had negative and 460 (22%) had positive resection margins. Having positive margins nearly doubled the risk of local recurrence and increased the risk of distant recurrence and disease related death. Seventy-two percent of patients with positive margins had no recurrence. Resection margin did not predict local control for retroperitoneal sarcomas or fibrosarcomas. Resection margin remained significantly associated with distant recurrence-free survival and disease-specific survival across all subsets after adjusting for other prognostic variables. The overall 5-year disease-specific survival rates for negative and positive margins were 83% and 75%. CONCLUSIONS: Positive microscopic resection margins significantly decrease the local recurrence-free survival rate for other-than-primary fibrosarcoma and retroperitoneal sarcomas, and independently predict distant recurrence-free survival rates and disease-specific survival rates for all patient subsets. Adjuvant therapy should be considered in the management of soft tissue sarcoma to increase local control. Because 72% of positive margins did not equate with inevitable local recurrence, considerable clinical judgment is required in considering additional treatment. Microscopic resection margins should be considered for inclusion in staging systems and treatment algorithms that address local recurrence. PMID- 11882766 TI - Isolated liver transplantation in infants with end-stage liver disease associated with short bowel syndrome. AB - OBJECTIVE: To evaluate experience with isolated orthotopic liver transplantation in children with liver failure associated with short bowel syndrome (SBS). SUMMARY BACKGROUND DATA: Infants who have liver failure as a result of SBS are frequently referred for consideration for combined liver and small bowel transplantation. In a few patients the liver disease develops despite a seemingly adequate bowel, which if given time and appropriate management has the potential for full enteral adaptation. There is a limited literature suggesting the utility of OLT without replacement of the native bowel. The advantages over combined liver and small bowel transplantation are clear: organ availability is greater, liver-reduction techniques are well established, lower immunosuppression is required, and there is greater experience in the care of children after orthotopic liver transplantation. METHODS: Eleven infants, considered to have a good prospect of eventual gut adaptation to full enteral nutrition if it were not for their advanced liver disease, underwent isolated orthotopic liver transplantation. Age range was 6.5 to 17.7 months. All patients had been dependent on parenteral feeding but had also shown significant enteral tolerance at some time before listing for transplantation. Advanced liver disease was apparent both clinically and on histologic examination. All were jaundiced and had low albumin levels, and most had coagulopathy. As a group the infants had growth retardation. Estimated remaining length of small bowel beyond the ligament of Treitz was in the range of 25 to more than 100 cm. Six infants retained their ileocecal valve. RESULTS: Thirteen liver transplants were performed in the 11 patients. A combination of whole livers (n = 6) and reduced-size grafts, of which three were from living-related donors, were used. Biliary anastomosis was duct-to duct in eight instances and involved a short Roux limb in the others. Eight patients are alive with follow-up of 15 to 66 months. Three deaths have occurred after transplantation as a result of sepsis. Of eight surviving patients, only two continue to receive intravenous support and in both there is increasing enteral tolerance. Since transplantation, all surviving children have shown adequate growth with maintenance of pretransplant centiles. CONCLUSIONS: In selected infants with liver failure secondary to short bowel syndrome in whom complete enteral autonomy is anticipated, isolated liver transplantation can offer long-term survival. PMID- 11882769 TI - Remember the distinction between malignant and benign pain? Well, forget it. PMID- 11882767 TI - Prognostic significance of cyclooxygenase 2 mRNA expression in non-small cell lung cancer. AB - OBJECTIVE: To investigate cyclooxygenase-2 (COX-2) mRNA expression in curatively resected non-small cell lung cancer (NSCLC) and to determine its association with prognosis. SUMMARY BACKGROUND DATA: Lung cancer is one of the most common malignancies in the world. Despite improvements in the diagnosis and treatment of NSCLC, the 5-year survival rate remains less than 15%. Identification of prognostic predictors based on molecular alterations could lead to additional diagnostic tools and eventually to more effective therapeutic options. Overexpression of COX-2 has been reported in several human malignancies, including lung cancer, but the prognostic importance of this overexpression has not been elucidated. METHODS: COX-2 mRNA expression was analyzed using a quantitative real-time polymerase chain reaction (Taqman) method in surgically resected tumor specimens from 89 patients with curatively resected NSCLC. RESULTS: COX-2 mRNA was detectable in all 89 (100%) tumor tissues. High COX-2 expression in tumors was significantly associated with inferior survival. Multivariate analysis showed that high COX-2 expression is an independent predictor of worse survival in patients with NSCLC. CONCLUSIONS: High COX-2 mRNA expression is an important biomarker for biologically aggressive disease in NSCLC and might be helpful in identifying patients who would benefit from additional therapies for controlling their disease. PMID- 11882770 TI - Major depression and insomnia in chronic pain. AB - OBJECTIVES: Insomnia and depression are common problems for people with chronic pain, and previous research has found that each is correlated with measures of pain and disability. The goal of this study was to examine the combined impact of major depression and insomnia on individuals with chronic pain. METHODS: The participants were patients with chronic musculoskeletal pain who underwent evaluation at an interdisciplinary treatment center. On the basis of semistructured interviews, participants were classified in three groups depending on whether they: (1) met criteria for major depression with insomnia (n = 38); (2) had insomnia without major depression (n = 58); or (3) had neither insomnia nor major depression (n = 47). The groups were then compared on self-report measures that included the McGill Pain Questionnaire, the Beck Depression Inventory, and the Multidimensional Pain Inventory. RESULTS: Participants with major depression and insomnia reported the most difficulty on measures of affective distress, life control, interference, and pain severity, although the insomniac patients without major depression also had elevated scores on some measures. In regression analyses, insomnia severity ratings did not contribute uniquely to the prediction of psychosocial problems when depression was controlled, but they did contribute to the prediction of pain severity. CONCLUSIONS: These results suggest that patients with chronic pain and concurrent major depression and insomnia report the highest levels of pain-related impairment, but insomnia in the absence of major depression is also associated with increased pain and distress. PMID- 11882771 TI - Evidence for the optimal management of acute and chronic phantom pain: a systematic review. AB - OBJECTIVES: The objective was to examine the evidence to determine the optimal management of phantom limb pain in the preoperative and postoperative phase of amputations. METHODS: Trials were identified by a systematic search of MEDLINE, review articles, and references of relevant trials from the period 1966-1999, including only English-language articles. Included trials involved a control group, any intervention, and reported phantom pain as an outcome. RESULTS: Twelve trials were identified, including 375 patients whose follow-ups ranged in duration from 1 week to 2 years. Only three randomized, controlled studies with parallel groups and three randomized crossover trials were identified. Eight trials examined treatment of acute phantom pain, including epidural treatments (three trials), regional nerve blocks (three trials), treatment with calcitonin (one trial), and transcutaneous electrical nerve stimulation (one trial). Three trials demonstrated a positive impact of the intervention on phantom limb pain, but the remainder demonstrated no difference between the intervention and control groups. Four trials examined late postoperative interventions, including transcutaneous electrical nerve stimulation (two trials) and the use of Farabloc (a metal threaded sock) and ketamine (one trial each). With regard to late postoperative interventions, three of the four trials showed modest short-term reduction of phantom limb pain. There was no relation between the quality of the trial and a positive result of the intervention. CONCLUSIONS: Although up to 70% of patients have phantom limb pain after amputation, there is little evidence from randomized trials to guide clinicians with treatment. Evidence on preemptive epidurals, early regional nerve blocks, and mechanical vibratory stimulation provides inconsistent support for these treatments. There is currently a gap between research and practice in the area of phantom limb pain. PMID- 11882772 TI - Complex regional pain syndrome type I: associated visual sensorimotor case findings. AB - OBJECTIVE: The objective was to present new visual sensorimotor findings in a patient with complex regional pain syndrome type I, formerly known as reflex sympathetic dystrophy. DESIGN: Clinical measurements were compared for the following visual sensorimotor tasks before and after 10 minutes of near visual stimulation: accommodation, vergence, and reading eye movements. PATIENT: The patient was a 19-year-old female university student with complex regional pain syndrome type I. RESULTS: All visual sensorimotor findings worsened dramatically after performance of the brief near visual task. In addition, the patient experienced severe dizziness, nausea, dull eye ache, and general fatigue, which persisted for 30 to 45 minutes following each test period. CONCLUSIONS: The patient manifested signs and symptoms of complex regional pain syndrome type I per the prior neurologic diagnosis, as well as the newly diagnosed accommodative infacility, accommodative insufficiency, convergence insufficiency, and deficits of saccades and pursuits, which were severely debilitating. The findings neither support nor refute the conventional notion of abnormal sympathetic mediation as a mechanism of fatigue and pain. However, the diagnoses of accommodative infacility and insufficiency suggest abnormal parasympathetic activation. Further investigation is needed to characterize the array of visual dysfunctions in a large sample of such patients, which may help elucidate the precise underlying neurologic causes of the sensorimotor deficits in these patients. PMID- 11882773 TI - Stability of pain parameters and pain-related quality of life in adolescents with persistent pain: a three-year follow-up. AB - OBJECTIVE: Many juveniles with chronic pain of no known organic cause recover. Because adolescents whose pain persists may have chronic pain as adults, a subsample of 42 adolescents from a prevalence study in which continuation of their pain was observed throughout the study period was investigated quantitatively and qualitatively. All mothers (n = 42) completed a questionnaire on the impact of the adolescent's pain on the family. The authors tested the hypothesis that pain parameters, pain-related quality of life, and impact of pain on the family would deteriorate over time. DESIGN: Three-year follow-up questionnaires, diaries, and interviews were used. SETTING: The study was conducted in the general population in the Rotterdam area. PARTICIPANTS: Adolescents (aged 12-18 years) who indicated chronic pain in our previous prevalence study and in a diary and questionnaire each year of the 3-year follow up were included in the study. RESULTS: The most prevalent pains were limb pain and headache. The pain intensity was mild (33 mm on a visual analog scale), very frequent (72% of all diary entries), and associated with relatively poor functional status and poor psychological and somatic functioning. The pain parameters, pain-related quality of life, and impact of pain on the family (i.e., restrictions in social life and problems in dealing with the stress of the adolescent's pain) remained surprisingly stable across the assessments. The interviews showed that pain had become part of the daily life of several adolescents, who structured their activities and sleeping hours to prevent aggravation of pain. In particular, adolescents with headache reported problems with cognitive activities, whereas those with limb pain and back pain reported problems with physical activities. CONCLUSIONS: For adolescents with persistent pain with no known organic cause, intensity and frequency of pain, quality of life, and impact of pain on the family did not change. Generally, they seemed to cope quite well with their pain. In view of these results, further studies should involve follow-up of adolescents with persistent pain into adulthood to establish the determinants of their pain and to find out whether they maintain their adaptive ways of living with their pain. PMID- 11882774 TI - Repeated assessment of temporomandibular joint pain: reasoned decision-making with use of unidimensional and multidimensional pain scales. AB - BACKGROUND: There are no reports in the literature about patients with painfully restricted temporomandibular joints concerning the reliability of unidimensional and multidimensional pain scales on the basis of a generalizability and decision study. Generalizability and decision studies are designed to aid in reasoned decision-making and therefore are complementary to classic psychometric analyses, in which correlation coefficients express the reliability of a measurement design. The smallest detectable difference as an outcome of the decision study is the smallest statistically significant change that can be detected. PATIENTS: Twenty-five consecutive patients (5 males and 20 females) with painfully restricted mandibular function participated in the current study. DESIGN: Repeated measurements of three levels of unidimensional experienced pain (average, minimal, and maximal pain before the week of assessment; n = 200) and the multidimensional pain rating index of the MPQ (n = 100) were used. The measurement sessions were supervised by two observers on two separate measurement days, 1 week apart, with each measurement repeated twice. RESULTS: Inconsistency in measurement results analyzed in terms of absolute error variance (i.e., the measurement facets plus all the interactions) represented at least 24% of total variance. The smallest detectable difference of visual analog scales varied from 43 to 15 mm, depending on the amount of repetitions, whereas for the pain rating index it varied from 22.7 to 14.4 units. CONCLUSIONS: For statistically and clinically successful treatment of patients with painfully restricted temporomandibular joints, clinicians must overcome at least the smallest detectable difference and 38% of the initial average pain level. PMID- 11882775 TI - Long-term use of gabapentin for treatment of pain after traumatic spinal cord injury. AB - OBJECTIVE: To determine the long-term efficacy of gabapentin as a treatment of pain after spinal cord injury. DESIGN: All patients with traumatic-onset spinal cord injury treated with gabapentin were identified and followed up using a longitudinal observational design with two contact points (6 and 36 months after the trial) using a semi-structured interview. The first follow-up interview attempted to capture all 31 patients placed on therapeutic trial. The second follow-up interview attempted to capture those reporting a favorable response (n = 14) to the therapeutic trial at the first follow-up. RESULTS: Of the 27 patients contacted at the first follow-up (87% response rate), 6 (22%) discontinued the trial secondary to intolerable side effects; therefore, the pain analgesic effects of gabapentin in these patients could not be determined. Of the remaining 21 patients, 14 (67%) reported a favorable response (i.e., a 2 or greater point reduction on a 0-10 pain-rating scale). The second follow-up interview captured 11 (79% response rate) of the 14 patients reporting a favorable response at the first interview, and 91% (10 of 11 patients) continued to report that gabapentin was an effective analgesic. There was no evidence to suggest dosing difficulties due to tolerance over the 3-year period. Sedation, dizziness, and forgetfulness were the most common side effects. CONCLUSIONS: Gabapentin may be an effective treatment of pain after spinal cord injury among those able to tolerate initial and long-term side effects. PMID- 11882776 TI - Painful polyneuropathy in patients with and without diabetes: clinical, neurophysiologic, and quantitative sensory characteristics. AB - OBJECTIVES: To study pain characteristics and peripheral nerve involvement in patients with painful diabetic and nondiabetic polyneuropathy in comparison with patients with non-painful polyneuropathy. PATIENTS AND METHODS: Fifty-five patients with polyneuropathy (37 with painful polyneuropathy, of whom 19 had diabetes and 18 had no diabetes; and 18 with painless polyneuropathy of different etiologies) were examined clinically using quantitative sensory tests and neurophysiology. Pain intensity and characteristics were analyzed by daily ratings on a 10-step verbal scale and by a questionnaire. RESULTS: Most patients experienced pain of more than one character. There was no clear difference in character or duration of pain between patients with and without diabetes. The mean value of the daily rating of pain intensity showed that pain was more severe in the evenings than in the mornings and that diabetic patients reported worse pain than nondiabetic patients. Thirty-two of the 37 patients with pain had paresthesias and/or dysesthesias, whereas only 7 of 18 patients without pain had paresthesias. Pain was always located in the feet, and, in most patients, also in the lower part of the legs. Some patients also experienced pain in the hands. Tactile sensibility, measured by quantitative tests, was more affected in both diabetic and nondiabetic patients with painful polyneuropathy compared with patients without pain (p = 0.02). Temperature, pain, and vibratory sensibility were equally affected in all patient groups. Nerve conduction velocity, amplitudes, and distal latency were equally affected in the pain group as compared with the control group, indicating that both thin and thick nerve afferents are affected in patients with painful as well as non-painful polyneuropathy and that etiology has no clear impact on nerve involvement. CONCLUSIONS: Neuropathy pain was always located in the feet and more severe in diabetic patients compared with patients with neuropathy pain of other etiologies. The authors also found evidence for a greater tactile sensibility involvement in patients with neuropathy pain, irrespective of etiology, whereas other quantitative sensibility and neurography parameters were equally affected in all patient groups. PMID- 11882777 TI - Management of progressive pain in a patient with intramedullary chordoma of the spine. AB - OBJECTIVES: The case here presented adequately reflects the difficulties involved in the treatment of pain in patients where the neuropathic component of pain predominates, and shows the different therapeutic steps that may be taken-from surgery and radiotherapy, to the administration of different drugs via the spinal route, to, finally, the presently little-used option of a direct intraventricular access. CONCLUSIONS: Spinal tumors are infrequent, but pose great difficulties for the management and control of the pain they cause. The utility of the spinal route as an early approach for the provision of adequate analgesia seems clear. However, it also appears to lose efficacy with time, and dose incrementing and/or the addition of drugs that enhance the analgesic action of morphine are not always effective. In such selected cases, the intraventricular route may constitute a useful alternative, allowing improved symptoms control with lower morphine doses, and the use of the system previously implanted for intrathecal spinal infusion. PMID- 11882778 TI - Meralgia paresthetica in differential diagnosis of low-back pain. AB - OBJECTIVE: Meralgia paresthetica is a syndrome of pain or dysesthesia or both in the anterolateral thigh, caused by entrapment of the lateral femoral cutaneous nerve at the anterior superior iliac spine. The aim of this report is to emphasize that meralgia paresthetica can be confused with low-back pain. PATIENT: A 21-year-old man was admitted to hospital because of low-back and thigh pain. He had a history of low-back pain. Physical examination and radiologic studies for low-back pain and radiculopathy showed no pathologic findings. It was suspected that the most likely cause was lateral femoral cutaneous neuropathy, caused by the wide military belt he continuously wore tightly around his waist. INTERVENTIONS: The nerve was blocked with 10 ml of bupivacaine 0.25%, which provided immediate pain relief. A nonsteroidal anti-inflammatory drug was administered orally. RESULTS: After 15 days of bed rest and 45 days without the belt, he was completely symptom-free. CONCLUSIONS: It is important to be rigorous in investigating the etiology of low-back pain. Meralgia paresthetica can mimic low-back pain because of the similarity of the symptoms. It can be treated by conservative or ablative therapeutic interventions; however, conservative methods should be considered primarily. PMID- 11882779 TI - Re: McCraken et al., A comparison of blacks and whites seeking treatment for chronic pain. Clin J Pain 2001;17:249-55. PMID- 11882780 TI - Will the learners be learned? PMID- 11882781 TI - A comprehensive review of genetic association studies. AB - Most common diseases are complex genetic traits, with multiple genetic and environmental components contributing to susceptibility. It has been proposed that common genetic variants, including single nucleotide polymorphisms (SNPs), influence susceptibility to common disease. This proposal has begun to be tested in numerous studies of association between genetic variation at these common DNA polymorphisms and variation in disease susceptibility. We have performed an extensive review of such association studies. We find that over 600 positive associations between common gene variants and disease have been reported; these associations, if correct, would have tremendous importance for the prevention, prediction, and treatment of most common diseases. However, most reported associations are not robust: of the 166 putative associations which have been studied three or more times, only 6 have been consistently replicated. Interestingly, of the remaining 160 associations, well over half were observed again one or more times. We discuss the possible reasons for this irreproducibility and suggest guidelines for performing and interpreting genetic association studies. In particular, we emphasize the need for caution in drawing conclusions from a single report of an association between a genetic variant and disease susceptibility. PMID- 11882782 TI - NAD(P)H:quinone oxidoreductase (NQO1) polymorphism, exposure to benzene, and predisposition to disease: a HuGE review. AB - NAD(P)H:quinone oxidoreductase (NQO1) catalyzes the two- or four-electron reduction of numerous endogenous and environmental quinones (e.g., the vitamin E alpha-tocopherol quinone, menadione, benzene quinones). In laboratory animals treated with various environmental chemicals, inhibition of NQO1 metabolism has long been known to increase the risk of toxicity or cancer. Currently, there are 22 reported single-nucleotide polymorphisms (SNPs) in the NQO1 gene. Compared with the human consensus (reference, "wild-type") NQO1*1 allele coding for normal NQO1 enzyme and activity, the NQO1*2 allele encodes a nonsynonymous mutation (P187S) that has negligible NQO1 activity. The NQO1*2 allelic frequency ranges between 0.22 (Caucasian) and 0.45 (Asian) in various ethnic populations. A large epidemiologic investigation of a benzene-exposed population has shown that NQO1*2 homozygotes exhibit as much as a 7-fold greater risk of bone marrow toxicity, leading to diseases such as aplastic anemia and leukemia. The extent of the contribution of polymorphisms in other genes involved in the metabolism of benzene and related compounds-such as the P450 2E1 (CYP2E1), myeloperoxidase (MPO), glutathione-S-transferase (GSTM1, GSTT1), microsomal epoxide hydrolase (EPHX1), and other genes-should also be considered. However, it now seems clear that a lowered or absent NQO1 activity can increase one's risk of bone marrow toxicity, after environmental exposure to benzene and benzene-like compounds. In cancer patients, the NQO1*2 allele appears to be associated with increased risk of chemotherapy-related myeloid leukemia. Many other epidemiological studies, attempting to find an association between the NQO1 polymorphism and one or another human disease, have now begun to appear in the medical literature. PMID- 11882783 TI - Needs assessment study of genetics education for general practitioners in Australia. AB - PURPOSE: Because of the explosion of genetic information resulting from the Human Genome Project and other developments in genetics and genetic technologies, primary care practitioners in Australia will be expected to have a much greater role in the practice of genetic medicine; however, little is published regarding their knowledge and attitudes to genetics, technologies, or genetics education. This study aimed to determine the genetics knowledge and educational needs of general practitioners (GPs) in Victoria, Australia, and their experiences in dealing with genetics in their practices. METHODS: Qualitative research methods were used primarily to obtain data from focus groups, interviews, and a limited survey. RESULTS: GPs believed their knowledge of genetics was poor and they felt inadequately prepared to manage patients with genetic conditions. They identified testing and counseling as areas that particularly needed strengthening. CONCLUSION: These data, together with GPs' suggestions about educational strategies, are essential for developing appropriate genetics education programs and resources that are relevant for Australian health care. PMID- 11882784 TI - Intergenerational transmission of pathogenic heteroplasmic mitochondrial DNA. AB - PURPOSE: To study the pattern of intergenerational transmission of pathogenic mitochondrial DNA with heteroplasmic A3243G, G8363A, A8344G, T8993G, and T8993C mutations. METHODS: The mutant load in the carrier mother and her offspring was measured in a total of 79 transmissions. Statistical analysis was performed to determine whether the intergenerational change in heteroplasmic mutant mtDNA is significant. RESULTS: Our results demonstrate that A3243G and T8993G mutant mtDNAs are significantly increased in blood, hair follicles, and buccal mucosal cells, during intergenerational transmission, whereas the intergenerational increase in T8993C and A8363G mutant mtDNA is not significant. Unlike previous reports, in one large family with A8344G mutation, the mutant load was slightly increased, instead of decreased, during transmission. There is no significant difference in the intergeneration transmission of mutant mtDNA to male or female offspring. CONCLUSION: Intergenerational transmission of heteroplasmic A3243G and T8993G mtDNA results in significant increase of mutant mtDNA in the offspring. Transmission of T8993C, G8363A, and A8344G does not result in significant intergenerational change in mutant load. Tissue specificity and genetic background may play important roles in the transmission of pathogenic heteroplasmic mtDNA. PMID- 11882786 TI - Updated assessment of cystic fibrosis mutation frequencies in non-Hispanic Caucasians. AB - PURPOSE: To update estimates of individual and cumulative cystic fibrosis (CF) mutation frequencies in non-Hispanic Caucasians for the prenatal screening panel recommended by American College of Medical Genetics and to determine the impact on screening performance. METHODS: Two data sources were used. In the first (CF Genetic Analysis Consortium), our re-analysis was restricted to North American studies. In the second (CF Foundation National Patient Registry), we performed a new analysis restricted to individuals tested at eight Therapeutic Development Network sites. RESULTS: The updated average cumulative proportion of mutations identified is 88.34% (higher than previously reported), indicating that 78% of high-risk couples (and affected fetuses) can potentially be identified. CONCLUSION: Prenatal CF screening in U.S. non-Hispanic Caucasians is more effective than previously thought. PMID- 11882788 TI - Theory and practice of imaging saturable binding sites. PMID- 11882785 TI - Barriers to successful dietary control among pregnant women with phenylketonuria. AB - PURPOSE: The teratogenic effects of maternal PKU are preventable, yet affected babies continue to be born. This study's purpose was to identify barriers to successful dietary control among pregnant women with PKU. METHODS: An interview based study was conducted of women with PKU who were known to metabolic disease clinics in three states and pregnant during 1998 to 2000. Medical records were used to document timing of metabolic control. RESULTS: Of 24 women in the study, only 8 (33%) initiated the diet before pregnancy. Of 22 medical records received, only 12 (55%) indicated control of blood phenylalanine levels before 10 weeks' gestation. Risk factors for late dietary control included young age and belief that treatment costs complicated the diet. Although all of the women expressed confidence in the metabolic clinic staff, few perceived their obstetricians were knowledgeable about the maternal PKU diet. Of 13 women enrolled in state-based assistance programs, 9 (69%) reported proof of pregnancy was required for eligibility. Many women using private insurance reported their insurers were unwilling to pay for medical foods. When the data were stratified according to state of residence, differences were observed in the rate of live-born infants, prepregnancy medical food use, average travel time to the metabolic clinic, and gestational week when metabolic control was achieved. CONCLUSION: Our study's findings may be used to target educational messages to women with PKU and to direct future research directions. For example, obstetric knowledge of maternal PKU needs further evaluation. Discrepancies should be resolved between maternal PKU medical recommendations and the policies of third party-payers. The disparities in financial assistance and services available to pregnant women with PKU residing in different states should be examined further. PMID- 11882789 TI - Using radiotracers to characterize magnetic resonance imaging contrast agents. PMID- 11882790 TI - Magnetic resonance imaging contrast enhancement of brain tumors at 3 tesla versus 1.5 tesla. AB - RATIONALE AND OBJECTIVES: To compare the diagnostic efficacy of a standard dose of MRI contrast agent in the evaluation of primary brain tumors and metastases using a high-field 3 tesla MR unit versus a 1.5 tesla MR unit. METHODS: Sixteen patients with brain tumors were examined at both field strengths using identical axial T1-SE protocols pre- and postcontrast (0.1 mmol/kg gadolinium), and postcontrast coronal 3D GRE with magnetization preparation (MP-RAGE), which was adjusted separately for each field strength. Evaluation of the images was performed quantitatively and, in the case of T1-SE images, also by visual assessment. RESULTS: Tumor-to-brain-contrast after gadolinium administration using statistical evaluation of MP-RAGE scans was significantly higher at 3 tesla (97.5) than at 1.5 tesla (46.3). The same was true for T1-SE sequences (93.0 vs. 72.1). Signal enhancement of the lesions in T1-SE sequences was not significantly different between both field strengths. CONCLUSIONS: Administration of a gadolinium contrast agent produces higher contrast between tumor and normal brain at 3 tesla than at 1.5 tesla. PMID- 11882791 TI - Gadolinium containing contrast agents for pulmonary ventilation magnetic resonance imaging: preliminary results. AB - RATIONALE AND OBJECTIVES: Five gadolinium containing contrast media (gadopentetate dimeglumine [Magnevist; Berlex Imaging, Montville, NJ, USA], gadobutrol [Gadovist; Schering, Jarfalla, Sweden], gadoteridol [ProHance; Bracco Byk Gulden, Konstanz, Germany], gadobenate dimeglumine [MultiHance; Bracco-Byk Gulden, Konstanz, Germany], and gadopentetate dimeglumine added with mannitol and a surface active detergent) were evaluated for their efficacy in magnetic resonance depiction of lung ventilation. METHODS: All contrast agent aerosols were generated by a jet nebulizer. Twelve intubated domestic pigs were mechanically ventilated with the respective aerosolized contrast agent and studied on a 1.5 T MR imager. T1-weighted TSE images using respiratory gating were obtained before and after a 10-minute ventilation period. Pulmonary signal intensity (SI) and signal-to-noise (SNR) changes were measured for both lungs. RESULTS: Mean SI increases ranged between 13.5% and 45.8% (right lung) and 14% and 39.8% (left lung). SNR changes ranged from +14.7% to +46.8% and from +13.1% to +40.5% for the right and left lung, respectively. The highest SI and SNR increases were observed in the gadoteridol group. CONCLUSIONS: The use of gadolinium for MR ventilation imaging is primarily hindered by its viscosity properties and thus, its capability of aerosolization. Of the five agents tested, the medium with the lowest viscosity at room temperature (gadoteridol) showed the most promising enhancement results. The results reaffirm the potential of gadolinium-based contrast agents as a pulmonary imaging alternative. With a reduction of ventilation duration down to ten minutes, the method appears tolerable in a clinical setting. PMID- 11882792 TI - Dynamic ventilation (3)He-magnetic resonance imaging with lung motion correction: gas flow distribution analysis. AB - RATIONALE AND OBJECTIVES: Software was developed to correct for lung motion to improve the description of hyperpolarized (3)He gas distribution in the lung. METHODS: Five volunteers were studied by dynamic ventilation (3)He-MRI using an ultrafast FLASH 2D sequence with a temporal resolution of 128 milliseconds. Signal kinetics were evaluated in the trachea and seven parenchymal Regions of Interest. Reference ranges for healthy subjects were defined for motion-corrected and uncorrected images. RESULTS: Motion correction was successfully performed. Reference ranges were 0.11-1.21 seconds for tracheal transit time, 0-0.02 seconds for trachea-alveolar interval, 0.22-0.62 seconds for alveolar rise time and 0 76.6 arbitrary units for alveolar amplitude for motion corrected images, and 0 1.09 seconds, 0-0.11 seconds, 0.26-0.85 seconds, 46.4-99.8 arbitrary units for uncorrected images. CONCLUSIONS: Evaluation of (3)He-distribution in the lung using motion correction of dynamic (3)He-ventilation imaging is feasible and gives more narrow reference ranges. PMID- 11882793 TI - Gadobenate dimeglumine in MRI of acute myocardial infarction: results of a phase III study comparing dynamic and delayed contrast enhanced magnetic resonance imaging with EKG, (201)Tl SPECT, and echocardiography. AB - RATIONALE AND OBJECTIVES: To evaluate the safety and utility of gadobenate dimeglumine as a magnetic resonance (MR) contrast agent in patients with acute myocardial infarction (MI). METHODS: One hundred three patients with acute MI received intravenous bolus gadobenate dimeglumine (0.05 mmol/kg) during MR examination. Dynamic and delayed T1-weighted spin-echo postcontrast images were compared with precontrast images, EKG, resting (201)Tl SPECT and echocardiography. RESULTS: Gadobenate dimeglumine was well tolerated. Dynamic imaging with gadobenate dimeglumine was more sensitive (72% vs 56%) than delayed spin echo imaging (P < 0.001). No difference in specificity was seen (98% vs 99%). (201)Tl SPECT was a sensitive (96%) test, but was not specific (63%). Echocardiography was not sensitive (32%), but was specific (92%). CONCLUSION: The intravenous use of gadobenate dimeglumine, at a bolus dose of 0.05 mmol/kg, is safe in patients with an acute MI. Dynamic contrast enhanced MR imaging has moderate sensitivity and high specificity for demonstrating infarct. PMID- 11882794 TI - Interstitial magnetic resonance lymphography using a polymeric t1 contrast agent: initial experience with Gadomer-17. AB - RATIONALE AND OBJECTIVES: The detection of lymph node metastases is an important step in tumor staging and is significant for therapy planning. Lymph node specific contrast agents can raise the sensitivity and specificity of modern diagnostic methods. This study investigated the suitability of the dendritic contrast agent Gadomer-17 in magnetic resonance (MR) lymph node imaging and compared three different dosages in such an application. METHODS: Doses of 1.0, 2.5, and 10.0 micromol Gd/kg body weight were interstitially injected into the hind legs of dogs; the signal intensities of two successive lymph node groups (inguinal and iliacal) were then recorded up to 120 minutes after injection. RESULTS: Gadomer-17 induced a strong increase in signal intensity of the examined lymph node groups. At 15 minutes postinjection, the enhancement increased by 120% to 680%, depending on the dose. The maximum enhancement was 450% to 960% at 60 to 90 minutes postinjection. Doses of 2.5 and 10.0 micromol Gd/kg showed comparable results; even the lowest dose (1.0 micromol Gd/kg) enhanced the contrast of the inguinal lymph nodes in 4 of 5 animals and the iliacal lymph nodes in three of five animals. Therefore, the minimum effective dose of Gadomer-17 in this study was approximately 2.5 micromol Gd/kg. CONCLUSION: This study revealed the excellent suitability of the dendritic contrast agent Gadomer-17 for MR imaging of the lymphatic system (lymph nodes and lymph vessels). PMID- 11882795 TI - Intrathecal gadolinium (gadopentetate dimeglumine) enhanced magnetic resonance myelography and cisternography: results of a multicenter study. AB - RATIONALE AND OBJECTIVES: This cooperative multicenter human study was designed to evaluate the safety, magnetic resonance (MR) imaging characteristics, and clinical response to a single gadolinium contrast agent: gadopentetate dimeglumine. MATERIAL AND METHODS: Ninety-five patients (age range: 1 month to 78 years; sex: 50 males, 45 females) were included in this prospective study. The patients presented clinically with a variety of cranial or spinal signs and symptoms for which an intrathecal contrast myelogram or cisternogram was requested by clinical staff. Via lumbar puncture (20-25 g needle), 3 to 5 mL/ml of cerebrospinal fluid were withdrawn and mixed with a single volume of 0.5 (n = 63), 0.7 (n = 13), 0.8 (n = 12), or 1.0 (n = 7) cc/mL of gadopentetate dimeglumine (Magnevist; Schering, Berlin, Germany). This was then injected into the subarachnoid space, and the needle was removed. Immediate and delayed (up to 96 hours) T1- and T2-weighted MR imaging was performed on super conductive, high field (1.0-1.5 tesla) imaging units in two or three planes. All patients were hospitalized for an observation period of 24 hours following the procedure, and follow-up neurologic examinations were performed serially for 6 to 12 months afterward. RESULTS: No patient manifested gross behavioral changes, neurologic alterations, or seizure activity at any time following the procedure. Nineteen patients (20%) experienced postural postlumbar puncture headache, six patients had nausea (6%), and two patients had episodes of vomiting (2%), all which resolved within the first 24 hours of the lumbar puncture with conservative bed rest. CONCLUSION: This cooperative study demonstrates the general safety and feasibility of low dose (0.5-1.0 mL/ml) intrathecal gadopentetate dimeglumine administration. The potential useful clinical applications include the evaluation of obstructions and communications of the various subarachnoid spaces, spontaneous or traumatic/postsurgical craniospinal cerebrospinal fluid leaks, and subarachnoid space CSF flow and parenchymal CNS interstitial diffusion dynamics. This worldwide cooperative study seeks to progressively perform human studies for further definitive evaluation of the practical clinical applications, of the relationship of this technique to other imaging studies and modalities, and the long-term safety of the procedure in a larger number of subjects. PMID- 11882796 TI - Changes in bone marrow-derived myeloid cells from thermally injured rats reflect changes in the progenitor cell population. AB - Bone marrow progenitor cells develop into mature tissue myeloid cells under the influence of colony-stimulating factors. Cytokines that are elevated post-thermal injury have been shown to influence this process. We hypothesize that thermal injury alters myelopoiesis at the level of the progenitor cell. These differences should be visible after in vitro cultures that include colony-stimulating factors. Prior to culture, bone marrow at postburn day 1 (PBD1) was assessed for cell surface markers and the levels of myeloid progenitors. After culture in granulocyte/macrophage-stimulating colony-stimulating factor, the cell surface markers of the cultured cells were determined. PBD1 marrow from thermally injured rats had more progenitor cells responsive to granulocyte/macrophage-stimulating colony-stimulating factor than did sham. Cultured PBD1 marrow produced more CD90(br) MY(br) CD45(dim) CD4(-) MHCII(-) CD11b(dim) eosinophils than did sham. Cultured bone marrow from thermally injured animals produces myeloid cells with an altered phenotype. Similar changes in myelopoiesis may take place in vivo. PMID- 11882797 TI - A multicenter review of toxic epidermal necrolysis treated in U.S. burn centers at the end of the twentieth century. AB - Toxic epidermal necrolysis (TEN) is a potentially fatal disorder that involves large areas of skin desquamation. Patients with TEN are often referred to burn centers for expert wound management and comprehensive care. The purpose of this study was to define the presenting characteristics and treatment of TEN before and after admission to regional burn centers and to evaluate the efficacy of burn center treatment for this disorder. A retrospective multicenter chart review was completed for patients admitted with TEN to 15 burn centers from 1995 to 2000. Charts were reviewed for patient characteristics, non-burn hospital and burn center treatment, and outcome. A total of 199 patients were admitted. Patients had a mean age of 47 years, mean 67.7% total body surface area skin slough, and mean Acute Physiology and Chronic Health Evaluation (APACHE II) score of 10. Sixty-four patients died, for a mortality rate of 32%. Mortality increased to 51% for patients transferred to a burn center more than one week after onset of disease. Burn centers and non-burn hospitals differed in their use of enteral nutrition (70 vs 12%, respectively, P < 0.05), prophylactic antibiotics (22 vs 37.9%, P < 0.05), corticosteroid use (22 vs 51%, P < 0.05), and wound management. Age, body surface area involvement, APACHE II score, complications, and parenteral nutrition before transfer correlated with increased mortality. The treatment of TEN differs markedly between burn centers and non-burn centers. Early transport to a burn unit is warranted to improve patient outcome. PMID- 11882798 TI - Venous thrombosis incidence in burn patients: preliminary results of a prospective study. AB - There are few prospective data on the incidence of deep venous thrombosis (DVT) in burn patients. In an on-going prospective study, hospitalized burn patients 18 years or older with an expected hospital length of stay more than 72 hours were imaged with baseline venous duplex ultrasound of all extremities within the first 48 hours after admission and weekly until discharge. Patient demographics and clinical risk factors for DVT were assessed. At the time of submission, 40 patients met screening criteria, and 30 were enrolled. Ultrasound diagnosed seven patients with 11 acute DVT for an incidence of 23%. One pulmonary embolism was documented. DVT patients had a mean age of 49 +/- 23 years with an average TBSA burn of 15 +/- 4% compared with those without thrombosis with a mean age of 44 +/ 17 years (P = NS) and TBSA burn of 18 +/- 25% (P = NS). There were no statistically significant differences for DVT patients in terms of age, number of central line days, hospital length of stay, or TBSA burned. Given the preliminary findings of this small study, we believe that all hospitalized burn patients are at risk for DVT. On-going investigation will be helpful in defining level of risk and improved prevention strategies for thromboembolic complications in burn patients. PMID- 11882799 TI - Long-term enhanced expression of heat shock proteins and decelerated apoptosis in polymorphonuclear leukocytes from major burn patients. AB - Heat shock proteins (HSPs), as molecular chaperones, have been reported to protect cells against a variety of environmental stresses. The objective of this study was to clarify the serial changes in expression of HSPs, oxidative activity, and apoptosis in polymorphonuclear leukocytes (PMNLs) from burn patients. Eight patients with severe burns (mean burn index 24.0 +/- 6.1) were included. Blood samples were serially obtained at five time points: days 0 to 1, days 2 to 7, days 8 to 14, days 15 to 21, and days 22 to 28. We measured expressions of HSP27, HSP60, HSP70, and HSP90 in permeabilized PMNLs by flow cytometry with the use of a monoclonal antibody against each HSP. The oxidative activity and apoptosis in PMNLs were also measured by flow cytometry. During all five time periods, expressions of HSP27, HSP60, and HSP70 in PMNLs from burn patients were significantly greater than those in PMNLs from healthy volunteers. The expression of HSP90 in PMNLs of burn patients increased between days 2 and 21. Oxidative activity in their PMNLs was significantly enhanced between days 2 and 28, and PMNL apoptosis was markedly inhibited for as long as 4 weeks after thermal injury. In conclusion, major burn causes long-term, enhanced expression of HSPs in PMNLs along with increased oxidative activity and decelerated apoptosis. The enhanced expression of HSPs may regulate the oxidative stress response and life-span of PMNLs in burn patients. PMID- 11882800 TI - An initial experience with telemedicine in follow-up burn care. AB - Following care in a burn unit, regular outpatient visits with burn specialists are required. The practical use of telemedicine in this context is unknown. The objective of this study was to evaluate patient and physician satisfaction with teleconsultations in follow-up burn care and to assess the costs and benefits of these teleconsultations. Fourteen teleconsultations were conducted between a burn physician and a patient at a remote site. Patients and the physician completed evaluation questionnaires for each teleconsultation. Time-related and financial costs of the consultation service were also determined. Patients were very satisfied with their teleconsultations and found them more economical and time efficient than in-person visits. The consulting physician felt teleconsultations were as satisfactory as clinic visits for the purposes of diagnosis and burn management. Our results support the ongoing use of telemedicine in the follow-up care of burn patients. PMID- 11882801 TI - Endogenous production of heparin-binding EGF-like growth factor during murine partial-thickness burn wound healing. AB - Heparin-binding EGF-like growth factor (HB-EGF), a potent epithelial cell mitogen, has been identified in human burn blister fluid and excised human burn wounds. Topical application of HB-EGF to murine partial-thickness scald burns accelerated reepithelialization, increased keratinocyte proliferation, and enhanced production of endogenous transforming growth factor-alpha in the healing wounds. The goal of the present study was to examine the production of endogenous HB-EGF and transforming growth factor-alpha (TGF-alpha) in a murine partial thickness scald burn model. Keratinocyte proliferation was assessed by 5-bromo deoxyuridine incorporation, and tissue sections were examined by in situ hybridization for HB-EGF mRNA expression and by immunohistochemistry for HB-EGF and TGF-alpha production. HB-EGF mRNA expression and production of HB-EGF and TGF alpha proteins by both marginal surface keratinocytes and hair follicle epithelial cells reached a maximum by postburn day five and decreased thereafter. This corresponded to the peak period of keratinocyte proliferation. We conclude that HB-EGF and TGF-alpha act in conjunction to stimulate wound healing following thermal injury. PMID- 11882802 TI - Differentiation and barrier formation of a cultured composite skin graft. AB - The differentiation and barrier formation of cultured composite skin grafts (CSGs) were assessed by histology and measurements of surface electrical capacitance (SEC) in vitro and in vivo. Keratinocytes cultured on the surface of acellular dermis were lifted to the air-liquid interface and analyzed for 30 days in vitro. Initially, SEC measurements of CSGs (n = 11) were high but quickly dropped between days 4 and 6 and remained steady for 30 days, indicating barrier formation by the epidermis. Histology of the CSGs (n = 6) demonstrated stratification of the epidermal cells and partial formation of the stratum corneum by day 3 that was complete by day 7. CSGs (n = 5) were transplanted to athymic mice, where they formed a stratified and differentiated epidermis. SEC measurements of CSGs remained low after transplant, suggesting that exposure to the air-liquid interface improved the maturation of CSGs in vitro prior to transplant. PMID- 11882803 TI - Burn centers should be involved in prevention of occupational electrical injuries. AB - Electrical injuries are uncommon, comprising 10% of our regional burn center admissions during a 9-year period. The purpose of this study was to determine the incidence, type, and location of occupation-related electrical injuries in an attempt to focus our injury prevention and outreach efforts. We retrospectively reviewed the medical records of patients with electrical injuries admitted to our burn center from January 1992 through March 2000, with focused analysis on those patients admitted with occupation-related electrical injuries. Of the 95 patients admitted for electrical burns, 81% (n = 77) were occupational injuries. This rate of injury suggests that prevention efforts should be directed at work sites and partnerships should be developed between burn centers and businesses to reduce the incidence of injuries. PMID- 11882805 TI - Irritable bowel syndrome. AB - Irritable bowel syndrome is a biopsychosocial disorder characterized by dysregulation of intestinal motor, sensory, and central nervous system functions. It is associated with significant disability and health-care costs. The traditional diagnostic approach leads to excessive investigation for many patients. A reductionist approach of focusing on evaluation and treatment of a single mechanism is unlikely to prove effective. Identification of the characteristic symptom pattern is the key to cost-effective management. A strong patient-physician relationship is essential for a successful outcome. Optimal outcome is predicated on an individualized treatment plan that integrates pharmacologic and behavioral modalities. PMID- 11882804 TI - Treatment of pain in acutely burned children. AB - The child with burns suffers severe pain at the time of the burn and during subsequent treatment and rehabilitation. Pain has adverse physiological and emotional effects, and research suggests that pain management is an important factor in better outcomes. There is increasing understanding of the private experience of pain, and how children benefit from honest preparation for procedures. Developmentally appropriate and culturally sensitive pain assessment, pain relief, and reevaluation have improved, becoming essential in treatment. Pharmacological treatment is primary, strengthened by new concepts from neurobiology, clinical science, and the introduction of more effective drugs with fewer adverse side effects and less toxicity. Empirical evaluation of various hypnotic, cognitive, behavioral, and sensory treatment methods is advancing. Multidisciplinary assessment helps to integrate psychological and pharmacological pain-relieving interventions to reduce emotional and mental stress, and family stress as well. Optimal care encourages burn teams to integrate pain guidelines into protocols and critical pathways for improved care. PMID- 11882807 TI - Profits and professionalism. PMID- 11882808 TI - QALYs or quackery? The quagmire of quantifying the cost of breathing. PMID- 11882809 TI - Molecular biologic substaging of non-small cell lung cancer. PMID- 11882810 TI - An economic evaluation of lung transplantation. AB - OBJECTIVE: This study was undertaken to determine the cost per quality-adjusted life-year gained with lung transplantation relative to medical treatment for end stage lung disease in the United Kingdom. METHODS: Patients on the transplant waiting list were used to represent medical treatment. Four-year national survival data were extrapolated to 15 years by means of parametric techniques. Quality-adjusted life-years were derived by means of utility scores obtained from a cross-section of patients. Resource consumption and costs were based on local and national sources. Costs and benefits were discounted at an annual rate of 6%. RESULTS: Across a 15-year period lung transplantation yielded mean benefits (relative to medical treatment) of 2.1, 3.3, and 3.6 quality-adjusted life-years for single-lung, double-lung, and heart-lung transplantation, respectively. During the same period the mean cost of medical treatment was estimated at $73,564, compared with $176,640, $180,528, and $178,387 for single-lung, double lung, and heart-lung transplantation, respectively. The costs per quality adjusted life-year gained were $48,241 for single-lung, $32,803 for double-lung, and $29,285 for heart-lung transplantation. Sensitivity analysis found the principal determinants of cost-effectiveness to be quality of life and maintenance costs after transplantation. CONCLUSIONS: Lung transplantation results in survival and quality of life gains but remains expensive, with cost effectiveness limited by substantial mortality and morbidity and high costs. The cost-effectiveness of lung transplantation can be improved with lowered immunosuppression costs and improvements in quality of life after transplantation. PMID- 11882811 TI - Marginal donor lungs: a reassessment. AB - OBJECTIVE: Lung transplantation is limited by the shortage of suitable donors. To overcome this problem, many programs have begun to use marginal or extended donors after reports suggesting equivalent outcomes with no additional risk. As our use of extended donor lungs increased and our recipient selection criteria expanded, we believed it was appropriate to reevaluate outcomes with extended donor lungs compared with outcomes with standard donor lungs and recipients outside of the currently accepted guidelines. METHODS: We performed a retrospective review of 128 consecutive lung or heart-lung transplants from January 1, 1997, to June 30, 2000. The primary endpoint was 30-day mortality. Donors were considered extended if any one of the following criteria were met: age greater than 55 years, smoking longer than 20 pack-years, presence of chest radiographic film infiltrate, PO (2) of less than 300 mm Hg, or purulent secretions on bronchoscopy. Guideline and nonguideline recipients were defined on the basis of previously published criteria. RESULTS: Of a total of 123 donors, 63 (51%) were extended. Forty-eight donors failed 1 criterion, 10 failed 2 criteria, and 5 failed 3 criteria. One hundred twenty-eight transplants were performed. The 30-day mortality for the standard donor group was 4 (6.2%) of 65 versus 11 (17.5%) of 63 for the extended donor group (P =.047). CONCLUSIONS: Although many extended donor lungs will result in acceptable postoperative function, caution needs to be exercised in the uses of certain extended donor lungs because there seems to be an increased early mortality rate in that group of recipients. Nonguideline recipients appear to have acceptable early mortality, except when they received extended donor lungs. PMID- 11882812 TI - Does donor cause of death affect the outcome of lung transplantation? AB - BACKGROUND: Accumulating evidence suggests that the donor's cause of death may influence posttransplantation allograft function. We conducted a retrospective analysis of our adult lung transplant experience to investigate the influence of donor traumatic brain injury versus nontraumatic brain injury on posttransplantation outcome. METHODS: We retrospectively reviewed donor records and recipient medical charts for 500 consecutive lung transplants performed between July 1988 and December 1999. Recipient follow-up was complete, with a minimum follow-up of 1 year of survival. RESULTS: There were 295 and 205 donors in the traumatic and nontraumatic brain injury groups, respectively. Young male donors predominated in the traumatic brain injury group. Recipients receiving donor lungs from the traumatic and nontraumatic brain injury groups did not differ by age, sex, diagnosis, type of transplant (single-lung versus double lung) or requirement for pretransplantation mechanical ventilatory assistance. Recipients did not differ in immediate or 24-hour PaO (2)/inspired oxygen ratio, ventilation time, hospital stay, hospital mortality, or overall survival. Recipients of organs from donors who died of traumatic brain injury showed a higher severity and frequency of rejection episodes during the first year after transplantation. Freedoms from bronchiolitis obliterans syndrome at 5 years were 34.5% and 50.8% for recipients of organs from donors who died of traumatic and nontraumatic brain injury, respectively (P =.002). CONCLUSIONS: The cause of donor brain death does not appear to influence early results of lung transplantation. Traumatic brain injury, or some phenomenon associated with it, may predispose a transplanted lung and its recipient toward more severe early rejection episodes and subsequent development of bronchiolitis obliterans syndrome. PMID- 11882813 TI - The Ross procedure is the procedure of choice for congenital aortic valve disease. AB - OBJECTIVES: The Ross procedure has emerged as an attractive option for aortic valve replacement in children and young adults. Our objective was to review our experience with the Ross procedure in young patients with congenital aortic valve disease. We also sought for evidence of growth in the autograft. METHODS: From January 1990 to July 2000, 260 patients underwent the Ross procedure for various aortic valve diseases. There were 136 patients less than 18 years of age. Fifty three (38%) of these patients had congenital aortic valve disease. Ages ranged from 3 months to 18 years (mean, 8 plus minus 5 years; median, 9 years). Ten patients were less than 2 years of age. Pure aortic stenosis was present in 18 patients, mixed stenosis and regurgitation in 32, and pure aortic regurgitation in 3. The aortic valve was bicuspid in 29 patients. Twenty-nine patients had previous procedures, mostly balloon dilation of the aortic valve (n = 8) or surgical aortic valvotomy (n = 12). RESULTS: In all patients immediate results demonstrated a normally functioning neoaortic valve with not more than trivial aortic valve regurgitation. In the patients with stenosis, all levels of obstruction were relieved, and the gradient across the left ventricular outflow tract was completely abolished. Hospital mortality was 3 (5.6%) of 53 (overall Ross mortality was 34 of 260 [1.5%]). The patients were followed up for a mean of 4 years and up to 10 years. One patient died late of a noncardiac cause. Actuarial survival at 10 years was 94% plus minus 2%, and freedom from all events was 93% plus minus 5%. Only 1 patient needed autograft replacement for endocarditis. Intervention related to right ventricle-pulmonary artery conduit was required in 3 patients: balloon dilatation in 2, and reoperation in 1. At last follow-up, all patients but one were classified as being in New York Heart Association functional class I or II with normal or near-normal autograft valve function. Serial measurement of the left ventricular outflow tract and aortic root showed that as patients grew, the size of the outflow tract increased. When indexed to body surface area, this increase correlated with the patients' expected somatic growth. CONCLUSIONS: The Ross procedure for congenital aortic valve disease in children and young adults offers excellent hemodynamics, with the added advantage of real potential for growth. It should be considered the treatment of choice in this age group. PMID- 11882814 TI - Aortic arch reconstruction with pulmonary autograft patch aortoplasty. AB - OBJECTIVE: The optimal technique for aortic arch reconstruction through median sternotomy is still under debate. We have introduced the technique of pulmonary autograft patch aortoplasty as a reliable alternative. METHODS: The outcomes of 51 infants who underwent neonatal repair of interrupted aortic arch (n = 28) or coarctation associated with ventricular septal defect (n = 23) since 1992 were analyzed. The patients were reviewed in three groups according to the aortic arch reconstruction technique: group I underwent direct anastomosis (n = 23), group II underwent homograft or pericardial patch aortoplasty (n = 8), and group III underwent pulmonary autograft patch aortoplasty (n = 20). The pulmonary autograft patch consisted in the anterior wall of the main pulmonary artery, between the supracommissural level and the divided ductus arteriosus. The created defect was replaced with fresh autologous pericardium. RESULTS: All patients except 1 were discharged without significant residual gradient at the level of the aortic arch. At a median delay of 7 months (range 2-51 months), 11 patients (22%) had recurrence of arch obstruction and underwent balloon angioplasty (n = 8) or surgical correction (n = 3). One patient who had undergone direct anastomosis required reoperation for bronchial compression. At a median follow-up of 29 months, the actuarial freedoms from recurrent arch obstruction were 81% for direct anastomosis, 28% for homograft or pericardial patch aortoplasty, and 100% for pulmonary autograft aortoplasty (P =.03 for group III vs group I and P <.0001 for group III vs group II). CONCLUSIONS: The aortic arch repair associated with pulmonary autograft patch augmentation resulted in superior midterm outcomes and therefore constitutes a reliable alternative to the direct anastomosis technique. It allowed complete relief of anatomic afterload and diminished the anastomotic tension, thus reducing the risk of restenosis and tracheobronchial compression. We observed a significantly higher rate of recurrence after patch aortoplasty with other materials. PMID- 11882815 TI - Newborn patients exhibit an unusual pattern of interleukin 10 and interferon gamma serum levels in response to cardiac surgery. AB - OBJECTIVE: The aim of this study was to determine the clinical significance of serum levels of interleukin 10 and interferon gamma in pediatric patients undergoing cardiopulmonary bypass. METHODS: We divided the patients into 2 groups: 8 neonates and 19 non-newborn children. Interleukin 10 and interferon gamma serum levels were quantified before sternotomy, at admission to the pediatric intensive care unit (30 minutes postoperatively), 24 hours after the onset of the operation, and 3 days after the operation. RESULTS: Newborn patients displayed significantly greater amounts of serum interleukin 10 than older children, not only in regard to the peak level achieved but also at every postoperative time point analyzed. In contrast, no significant changes in interferon gamma serum levels were observed in neonates at any time point, whereas non-newborn pediatric patients showed a significant increase in interferon gamma serum concentrations immediately after the operation. This unusual pattern of cytokine response in newborn patients was not associated with modifications in cortisol serum levels. Furthermore, although neonates had significantly different surgical and clinical variables than did the non-newborn pediatric patients, the variation in interleukin 10 production in neonates could not be accounted for by differences in the magnitude of surgical injury. In the group of neonates, there were significant positive correlations between peak interleukin 10 serum levels and both partial pressure of arterial oxygen/fraction of inspired oxygen ratio and postoperative body weight gain. CONCLUSIONS: Newborn patients undergoing cardiopulmonary bypass exhibit a distinctive biologic response pattern characterized by high levels of serum interleukin 10 without changes in serum interferon gamma. This cytokine imbalance could have potential clinical implications. PMID- 11882816 TI - Coagulation abnormalities in patients with single-ventricle physiology precede the Fontan procedure. AB - OBJECTIVE: Thromboembolic events in patients who have undergone the Fontan operation have been reported to be as high as 20% to 33%. A hypercoagulable state with deficiencies in proteins C and S has been implicated. Using age-matched control subjects, we evaluated whether an altered coagulation state is present earlier in the course of staged single-ventricle repair. METHODS: After informed consent had been obtained, coagulation factors were assayed in 36 infants (mean age, 7.7 +/- 3.6 months) with single-ventricle cardiac defects immediately before undergoing the bidirectional Glenn procedure; 34 infants (mean age, 8.4 +/- 2.6 months) without cardiac disease were assayed as control subjects. Concentration of factors II, V, VII, VIII, IX, and X; antithrombin III; plasminogen; proteins C and S; fibrinogen; serum albumin; and liver enzymes were measured. Normal reference intervals on the basis of the control subjects were determined by using 95% confidence limits. Patient demographic and hemodynamic variables were evaluated as possible predictors of coagulation abnormalities. RESULTS: Concentrations of protein C; factors II, V, VII, IX, and X; plasminogen; fibrinogen; and antithrombin III were significantly lower in the pre-Glenn infants compared with the age-matched control subjects (all P <.001, Student t test). On the basis of multiple logistic regression, no specific hemodynamic variables were predictive of a procoagulant or anticoagulant deficiency. Ventricular dysfunction did predict the presence of multiple coagulation abnormalities (P <.001). CONCLUSION: Procoagulant and anticoagulant factor abnormalities occur early in the course of single-ventricle repair and precede the cavopulmonary connection. PMID- 11882817 TI - Molecular assessment of lymph nodes in patients with resected stage I non-small cell lung cancer: preliminary results of a prospective study. AB - OBJECTIVES: Routine histologic examination of resected lymph nodes in patients with stage I non-small cell lung cancer may underestimate the incidence of advanced disease. The presence of occult lymph node metastases may predict a higher risk of recurrence after intended curative resection. The purpose of this study was to determine the prognostic significance of TP53 and K-ras mutations in histologically determined negative lymph nodes from patients with stage I non small cell lung cancer who underwent intended curative surgical resection. METHODS: Between July 1995 and March 1998, clinical data and tissue samples of primary tumors and lymph nodes were collected in a prospective fashion from 102 patients undergoing resection for non-small cell lung cancer (stage I, n = 55; stage II, n = 32; stage IIIA, n = 15). TP53 and K-ras mutations were detected by direct sequencing. If molecular alterations were found in the primary tumor, the corresponding lymph nodes were examined for these same TP53 (by oligonucleotide hybridization) and K-ras (by allele-specific ligation) mutations. RESULTS: TP53 mutations were found in 47 of 94 primary tumors (50%), and K-ras mutations were present in 26 of 55 adenocarcinomas (47%). A total of 134 lymph nodes from 32 patients with stage I disease were analyzed. In 9 cases (28%) the same TP53 or K ras mutations were found in tumor and lymph node specimens, suggesting occult metastasis. On the basis of nodal location, 7 patients had their disease upstaged by a single stage and 2 patients by two stages. All 28 patients with stage II or III disease had pathologically determined positive nodes that were confirmed as positive by molecular analysis. Standard histopathologic assessment of regional lymph nodes failed to detect metastases at levels below 0.9% tumor-specific mutant TP53 clones per node. No statistically significant difference in disease specific or overall survival was observed between patients with stage I disease with and without molecular lymph node metastases. CONCLUSIONS: Occult lymph node metastases are present in a significant percentage of patients with stage I non small cell lung cancer. These data suggest that molecular analysis allows a more accurate assessment of staging. However, larger studies are needed to determine the clinical role of molecular staging. PMID- 11882818 TI - Rapid, quantitative reverse transcriptase-polymerase chain reaction: application to intraoperative molecular detection of occult metastases in esophageal cancer. AB - OBJECTIVE: Our earlier data showed that quantitative reverse transcriptase polymerase chain reaction can discriminate patients with node-negative cancer who are at high risk for recurrence. The objective of this study was to determine whether a new, more rapid quantitative reverse transcriptase-polymerase chain reaction assay could provide this information in a time frame suitable for intraoperative decision making. METHODS: We studied formalin-fixed, archived lymph nodes from 30 patients with histologically determined node-negative esophageal cancer with rapid quantitative reverse transcriptase-polymerase chain reaction to measure expression of carcinoembryonic antigen messenger RNA. We also performed rapid quantitative reverse transcriptase-polymerase chain reaction on 37 snap-frozen lymph nodes from 23 patients. Eleven of the 23 patients had benign esophageal disorders (negative control group). The other 12 had esophageal cancer, 6 with histologically determined positive lymph nodes and 6 with histologically determined negative lymph nodes. RESULTS: In the retrospective analysis of archival tissue from 30 patients with esophageal cancer with histologically determined negative lymph nodes, rapid quantitative reverse transcriptase-polymerase chain reaction predicted disease recurrence with a sensitivity and a specificity of 90% and 80%, respectively, and was comparable to conventional quantitative reverse transcriptase-polymerase chain reaction. In the frozen-tissue analysis rapid quantitative reverse transcriptase-polymerase chain reaction detected significantly higher levels of carcinoembryonic antigen expression in all 12 of the histologically determined positive lymph nodes than in the benign nodes. For 2 of these 12 nodes the intraoperative frozen-section analysis had negative histologic results, and N1 status was determined only on final pathologic examination. Rapid (intraoperative) quantitative reverse transcriptase-polymerase chain reaction discriminated both nodes as positive. Among the 14 histologically determined negative nodes, 1 of 3 nodes from 1 patient showed increased carcinoembryonic antigen according to rapid quantitative reverse transcriptase-polymerase chain reaction, and this patient had a clinical recurrence. CONCLUSIONS: In our study we were able to rapidly discriminate patients with node negative-esophageal cancer who had a high risk of recurrence. In frozen tissues rapid quantitative reverse transcriptase-polymerase chain reaction correlated with final pathologic report for 11 of 12 patients. In the 1 discordant case, the quantitative reverse transcriptase-polymerase chain reaction result was positive and may have detected microscopically occult metastasis, because this patient did have disease recurrence. Rapid quantitative reverse transcriptase-polymerase chain reaction was more sensitive than intraoperative frozen sections for detecting metastatic disease. These data suggest that rapid quantitative reverse transcriptase-polymerase chain reaction may have a prognostic role and could guide intraoperative decisions. PMID- 11882819 TI - Molecular staging of lung cancer: real-time polymerase chain reaction estimation of lymph node micrometastatic tumor cell burden in stage I non-small cell lung cancer--preliminary results of Cancer and Leukemia Group B Trial 9761. AB - OBJECTIVE: The 5-year survival for patients with surgically resected stage I non small cell lung cancer is only 60% to 70%, probably because of undetected systemic occult micrometastases. Detection of occult micrometastases in lymph nodes by reverse-transcriptase polymerase chain reaction for carcinoembryonic antigen messenger RNA in non-small cell lung cancer has not been reported. Detection of occult micrometastases by standard reverse-transcriptase polymerase chain reaction provides only yes or no answers about their presence, whereas quantitative real-time reverse-transcriptase polymerase chain reaction permits reproducible quantitation of target molecules. This study evaluated the ability of quantitative reverse-transcriptase polymerase chain reaction to quantitate lymph node occult metastases with carcinoembryonic antigen messenger RNA as a tumor marker. METHODS: Standard reverse-transcriptase polymerase chain reaction and quantitative reverse-transcriptase polymerase chain reaction for carcinoembryonic antigen messenger RNA were performed on 232 lymph nodes from 53 patients with stage I disease (node negative according to histologic examination). Quantitative reverse-transcriptase polymerase chain reaction determined carcinoembryonic antigen messenger RNA quantity by detecting fluorescence increase at a threshold polymerase chain reaction cycle. Threshold polymerase chain reaction cycle values were correlated with standard curves created from serially diluted carcinoembryonic antigen-positive HTB-174 tumor cells to estimate the number of micrometastatic tumor cells in a lymph node. RESULTS: Detection rates of occult metastases were similar for standard reverse transcriptase polymerase chain reaction and quantitative reverse-transcriptase polymerase chain reaction at 38 of 232 (16.4 %) and 59 of 232 (25.4 %), respectively. Upstaging rates among 53 cases of stage I non-small cell lung cancer were also similar for standard reverse-transcriptase polymerase chain reaction and quantitative reverse-transcriptase polymerase chain reaction at 23 of 53 (43.4 %) and 30 of 53 (56.6%), respectively. Comparison of positive lymph node stations according to quantitative reverse-transcriptase polymerase chain reaction (threshold polymerase chain reaction cycle <45) with HTB-174 tumor cell standard curves yielded estimates of metastatic tumor cell burden of 1.07 x 10(3)to 3.24 x 10(5)cells per lymph node station (median 7190 tumor cells per lymph node station). CONCLUSIONS: Standard and quantitative real-time reverse transcriptase polymerase chain reaction for carcinoembryonic antigen detected occult metastases in patients with stage I non-small cell lung cancer at similar rates; both upstaged about 50% of cases. Quantitative reverse-transcriptase polymerase chain reaction allows estimation of the number of metastatic cells per lymph node, however, which potentially allows greater precision in predicting recurrence risk. PMID- 11882820 TI - Pleural complications of hydatid disease. AB - OBJECTIVE: The aim of this retrospective study was to review pleural and pericardial complications of patients with hydatid cysts and to analyze the management of therapy for these patients. METHODS: Between 1991 and 2001, 43 (29.7%) of 145 patients presented with pleural and pericardial complications. These patients had spontaneous pneumothorax (6.2%), empyema (7.6%), pleural thickening (10.3%), hepatopleural fistula (2.8%), pericarditis (2.1%), and hepatobronchial fistula (0.7%). There were 22 male and 21 female patients, with a mean age of 30 years. RESULTS: The most common symptom was chest pain (79.1%). In 37 (86.1%) of 43 patients, the cysts were unilateral. The ratio of ruptured cysts was 88.4%. In most of the patients, hydatid cysts developed in the right lung (62.9%) and the lower lobes of the lung (70.4%). Multiple cysts were found in 8 (18.6%) patients. The most common surgical techniques were cystotomy with capitonnage (55.7%) and decortication (69.8%). Radical lung resection was used in 14% of the patients. The morbidity rate was 16.3%, and the mortality rate was 2.3%. The mean follow-up was 19 months with no recurrence. CONCLUSIONS: Although lung-preserving surgical interventions should be preferred, radical surgical procedures have been used more commonly in patients with pleural complications of hydatid cysts, and the postoperative morbidity rate was higher in these patients. Because of this, the surgical treatment should be carried out before the development of pleural complications. In addition, echinococcosis should be considered and included in the differential diagnosis of spontaneous pneumothorax and empyema. PMID- 11882821 TI - Anatomic variations of the T2 nerve root (including the nerve of Kuntz) and their implications for sympathectomy. AB - OBJECTIVE: The aim of this study was to clarify the anatomic variations of the intrathoracic nerve of Kuntz, and this should help delineate the resection margins during video-assisted thoracic sympathectomy. METHODS: Sixty-six thoracic sympathetic chains of 39 adult Korean cadavers were dissected on both sides of the thorax in 27 cadavers (54 sides) and on one side in 12 cadavers (12 sides). RESULTS: The intrathoracic nerve was observed in 45 (68.2%) sides and was present bilaterally in 48.1% of cadavers. No intrathoracic nerve or ascending ramus communicans arising from the second thoracic nerve was observed in only 5 (7.6%) sides. The diameter of the intrathoracic nerve was 1.25 plus minus 0.55 mm on average. The arising point of the intrathoracic nerve from the second thoracic nerve was 7.3 mm on average from the sympathetic trunk. Presence of the stellate ganglion was noted in 56 (84.8%) sides, and 6 (9.1%) sides showed a single large ganglion formed by the stellate and the second thoracic sympathetic ganglia. The second thoracic sympathetic ganglion was most commonly located (50%) in the second intercostal space. CONCLUSION: The anatomic variations of the intrathoracic nerve of Kuntz and the second thoracic sympathetic ganglion were characterized in human cadavers. It is hoped that this study will help to improve the recurrence of symptoms caused by the intrathoracic nerve in an upper thoracic sympathectomy for hyperhidrosis. PMID- 11882822 TI - The significance of E-cadherin and alpha-, beta-, and gamma-catenin expression in surgically treated non-small cell lung cancers of 3 cm or less in size. AB - OBJECTIVES: Expression of the cell-cell adhesion molecules E-cadherin and alpha-, beta-, and gamma-catenin seems closely related to tumor invasiveness. The relationship between the expression and clinicopathologic characteristics in surgically resected non-small cell lung cancers of 3 cm or less in size was studied. The relationship to patient survival was analyzed. METHODS: A total of 115 patients with surgically resected lung cancers of 3 cm or less in size were enrolled in this study. Expression of E-cadherin and alpha-, beta-, and gamma catenin was immunohistochemically measured. The chi(2) test was used to correlate this expression with clinicopathologic parameters. Their influence on patient survival was evaluated with the Cox proportional hazards model. RESULTS: There was a positive correlation between E-cadherin and catenin expression in lung cancers. In general, E-cadherin and catenin expression were greater in tumors that were either bronchioloalveolar carcinomas or adenocarcinomas, well differentiated, early stage, peripheral, and without vascular or pleural invasion. By using multicovariate analysis of patient survival, only early-stage and peripheral tumors were significantly favorable prognostic factors. Further analysis of the group of patients with early-stage disease showed that higher alpha-, beta-, or gamma-catenin expression was a favorable prognostic indicator. CONCLUSION: Expression of alpha-, beta-, or gamma-catenin can be used as a prognostic indicator in patients with surgically resected stage I non-small cell lung cancers of 3 cm or less in size. PMID- 11882823 TI - Decreased sternal vascularity after internal thoracic artery harvesting resolves with time: an assessment with single photon emission computed tomography. AB - OBJECTIVE: We sought to prospectively evaluate the long-term effect of left internal thoracic artery harvesting on sternal vascularity. METHODS: Twelve consecutive patients undergoing primary coronary artery bypass grafting were studied. In all patients a pedicled left internal thoracic artery was harvested. Each patient underwent a preoperative technetium-99m methylene diphosphonate bone scan with single photon emission computed tomography. The ratio of the mean counts per pixel for each side of the sternum was obtained. All patients had early repeat bone scans 6 plus minus 2.4 days postoperatively and late repeat bone scans 18.6 plus minus 3.5 months after the operation. Ratios of unilateral sternal uptakes were compared between the different scans. One patient died during follow-up and was excluded from the study. RESULTS: There was a significant decrease in flow to the left hemisternum in the early postoperative scan compared with that in the preoperative scan (P <.001). At late follow-up scans, flow to the left hemisternum had returned to normal (P =.119). Midterm clinical follow-up demonstrated 3 superficial wound infections. No musculoskeletal pain existed at the time of follow-up, but 3 patients had numbness or tingling at the skin area corresponding to the site from which the left internal thoracic artery was harvested. CONCLUSIONS: Acute postoperative sternal ischemia caused by harvesting of a pedicled left internal thoracic artery is temporary and resolves with time. PMID- 11882824 TI - Effect of radial artery harvest on collateral forearm blood flow and digital perfusion. AB - BACKGROUND: Changes describing digital and forearm circulation after radial artery harvest have been reported infrequently. METHODS: This prospective study examined digital perfusion and forearm collateral circulation preoperatively and postoperatively in patients who underwent coronary artery bypass grafting with radial artery free grafts. Noninvasive evaluation was conducted with digital photoelectric plethysmography and color flow and pulsed Doppler studies. RESULTS: Thumb perfusion index decreased from 1.25 to 0.84 (30%, P <.001) in the unoperated extremities and from 1.23 to 0.80 (36%) in the operated extremities (P <.001). Doppler studies in extremities after radial artery harvest demonstrated an increase in ulnar artery diameter from 3.87 to 4.66 mm (15.7%, P <.001) and a rise in ulnar blood flow velocity from 38.96 to 48.46 cm/s (17.4%) preoperatively to 8 weeks postoperatively (P <.001). No hand ischemia was noted. CONCLUSIONS: Our study identified a mild reduction in digital perfusion and an increase in ulnar artery flow velocity and diameter with no clinical sequelae or compromise in hand function after radial artery harvest in properly selected patients. PMID- 11882825 TI - Effect of perioperative complications on excess mortality among women after coronary artery bypass: the Israeli Coronary Artery Bypass Graft Study (ISCAB). AB - BACKGROUND: Widely observed excess mortality among women after coronary artery bypass grafting is still largely unexplained, although case-mix factors have been identified. We evaluated the contribution of perioperative complications to the risk of 180-day mortality among women while adjusting for case-mix factors. METHODS: This is part of a prospective, 1-year nationwide Israeli coronary artery bypass graft study of 1029 female and 3806 male patients. Deaths within 180 days were independently ascertained. Case-mix risk strata were obtained from a pooled Cox survival model (including all subjects and study variables) by using the adjusted coefficients corresponding to the case-mix factors within the model. Sex specific mortality associated with perioperative complications was evaluated within the strata. In addition, sex-specific Cox models were constructed. RESULTS: Higher mortality among women compared with that among men was significant within the pooled model (hazard ratio, 1.4; P =.038) and was evident early in the postoperative period. Women tended to cluster in the highest risk quartile compared with men (39.8% vs 20.9%, P <.001). However, although the incidence of perioperative complications was similar for the 2 sexes, the associated mortality for a given perioperative complication was higher among women. Sex-specific Cox models confirmed the above findings. For example, the hazard ratio for women with low postoperative hemoglobin was 6.9, whereas for men, the hazard ratio was 3.9. CONCLUSIONS: The role of perioperative factors in the excess mortality among women after coronary artery bypass grafting shifts the focus of attention from the selection of women for the operation to the in hospital experience. Improving the outcome for women will entail efforts to prevent complications in the perioperative period. PMID- 11882826 TI - The early clinical and angiographic outcome of sequential coronary artery bypass grafting with the off-pump technique. AB - OBJECTIVE: The emergence of the off-pump coronary artery bypass technique has made surgeons consider combining it with other techniques developed and learned with conventional coronary artery bypass grafting with cardiopulmonary bypass. One of these techniques is the construction of a sequential graft to bypass more than one coronary vessel. The purpose of this study is to review the outcome of combining sequential coronary artery bypass grafting with off-pump techniques. METHODS: We retrospectively reviewed the records of 45 consecutive patients who underwent isolated coronary bypass surgery with off-pump and sequential grafting techniques at Harefield Hospital (Harefield, UK) between July 1999 and December 2000. The registry database, medical notes and charts were studied for preoperative and postoperative data of the patients. Ten patients consented and underwent early postoperative angiography to check the quality of the grafts and anastomoses. RESULTS: There were no deaths among the study patients. Morbidity consisted of atrial fibrillation in 6 patients (13.3%), leg wound infection in 2 patients (4.4%), and pleural effusion in 1 patient (2.2%). Early angiography of the 10 consenting patients revealed 10 patent sequential grafts (100%) with 20 satisfactory end-to-side and side-to-side anastomoses (100%). CONCLUSION: The combination of sequential grafting and off-pump techniques is feasible, is safe, and provides good early clinical and angiographic outcomes. PMID- 11882827 TI - Delayed detection of motor pathway dysfunction after selective reduction of thoracic spinal cord blood flow in pigs. AB - OBJECTIVE: Clinical monitoring of myogenic motor evoked potentials to transcranial stimulation provides rapid evaluation of motor-pathway function during surgical procedures in which spinal cord ischemia can occur. However, a severe reduction of spinal cord blood flow that remains confined to the thoracic spinal cord might render ischemic only the descending axons of the corticospinal pathway. In this situation lower-limb motor evoked potentials could respond relatively late compared with a similar spinal cord blood flow reduction of the lumbar spinal cord that renders predominantly motoneurons ischemic. METHODS: Selective thoracic and lumbar spinal cord ischemia was induced by sequential clamping of segmental arteries during continuous assessment of laser-Doppler spinal cord blood flow at the thoracic and lumbar spinal cord. Myogenic motor evoked potentials were recorded from the upper and lower limbs. The time to loss of motor evoked potentials was compared (n = 11) during reduction of laser Doppler spinal cord blood flow below 25% of baseline (ischemic segment), and flow was maintained at greater than 75% of baseline in the nonischemic segment, both during thoracic and lumbar spinal cord ischemia. RESULTS: Average laser-Doppler spinal cord blood flow in the ischemic segment was similar during thoracic (26% +/- 15% [+/- SD]) and lumbar (26% +/- 16%) ischemia, whereas normal flow was maintained in the nonischemic segment. The time to motor evoked potentials loss was considerably longer after thoracic spinal cord ischemia (15 +/- 11 minutes) than after lumbar spinal cord ischemia (3 +/- 2 minutes, P <.005). CONCLUSION: In this experimental model of selective spinal cord ischemia, a severe reduction of lumbar spinal cord blood flow results in rapid loss of myogenic motor evoked potentials, whereas a similar blood flow reduction in the thoracic spinal cord results in relatively slow loss of motor evoked potentials. The effectiveness of motor evoked potentials to rapidly assess spinal cord integrity might be limited when spinal cord ischemia is confined to the thoracic segments. PMID- 11882828 TI - Does tranexamic acid reduce desmopressin-induced hyperfibrinolysis? AB - OBJECTIVE: Desmopressin releases tissue-type plasminogen activator, which augments cardiopulmonary bypass--associated hyperfibrinolysis, causing excessive bleeding. Combined use of desmopressin with prior administration of the antifibrinolytic drug tranexamic acid may decrease fibrinolytic activity and might improve postoperative hemostasis. METHODS: This prospective randomized study was carried out with 100 patients undergoing coronary artery bypass operations between April 1999 and November 2000 in Gulhane Military Medical Academy. Patients were divided into 2 groups. Desmopressin (0.3 microg/kg) was administrated just after cardiopulmonary bypass and after protamine infusion in group 1 (n = 50). Both desmopressin and tranexamic acid (before the skin incision at a loading dose of 10 mg/kg over 30 minutes and followed by 12 hours of 1 mg.kg(-1).h(-1)) were administrated in group 2 (n = 50). RESULTS: Significantly less drainage was noted in group 2 (1010 +/- 49.9 mL vs 623 +/- 41.3 mL, P =.0001). Packed red blood cells were transfused at 2.1 +/- 0.5 units per patient in group 1 versus 0.9 +/- 0.3 units in group 2 (P =.0001). Fresh frozen plasma was transfused at 1.84 +/- 0.17 units per patient in group 1 versus 0.76 +/- 0.14 units in group 2 (P =.0001). Only 24% of patients in group 2 required donor blood or blood products compared with 74% of those in the isolated desmopressin group (group 1, P =.00001). Group 1 and group 2 findings were as follows: postoperative fibrinogen, 113 +/- 56.3 mg/dL versus 167 +/- 45.8 mg/dL (P =.0001); fibrin split product, 21.2 +/- 2.3 ng/mL versus 13.5 +/- 3.4 ng/mL (P =.0001); and postoperative hemoglobin level, 7.6 plus minus 1.2 g/dL versus 9.1 plus minus 1.2 g/dL (P =.0001). CONCLUSION: Tranexamic acid administration significantly reduces desmopressin and bypass-induced hyperfibrinolysis. Combined use of tranexamic acid and desmopressin decreases both postoperative blood loss and transfusion requirement. PMID- 11882829 TI - Induction and maintenance of an experimental model of severe cardiomyopathy with a novel protocol of rapid ventricular pacing. AB - OBJECTIVE: An animal model of chronic severe heart failure is needed to evaluate new mechanical devices, surgical procedures, and medical therapies. The purpose of this study was to evaluate a unique new model of severe heart failure developed by means of a novel protocol of rapid ventricular pacing. METHODS: Heart failure was induced in 8 mongrel dogs by means of rapid ventricular pacing (230 beats/min) for 4 weeks. After a sham operation, maintenance pacing at a reduced rate (190 beats/min) was continued for another 4 weeks. RESULTS: Left ventricular systolic function was significantly reduced at week 4 and remained low at week 8, including the slope of the end-systolic pressure-volume relationship (2.4 +/- 1.0 vs 0.7 +/- 0.2 vs 0.8 +/- 0.3 mm Hg/mL [baseline vs week 4 vs week 8, respectively]), ejection fraction (63% +/- 5% vs 28% +/- 7% vs 33% +/- 5%), and cardiac output (3.1 +/- 0.7 vs 2.0 +/- 0.3 vs 2.2 +/- 0.7 L/min). Significant ventricular remodeling changes took place with increased ventricular volumes and circumferential wall stress, which were stable between weeks 4 and 8. Serum catecholamine and atrial natriuretic polypeptide levels also increased from baseline but stabilized between weeks 4 and 8. The end-diastolic pressure-volume relationship also showed stable diastolic function between weeks 4 and 8. CONCLUSIONS: Induction pacing at 230 beats/min readily created severe heart failure in all animals, and a new technique of maintenance pacing provided a consistent model of severe heart failure. This model can be used to study a variety of new interventions for heart failure. PMID- 11882830 TI - Intravascular hemolysis in patients with new-generation prosthetic heart valves: a prospective study. AB - OBJECTIVE: A prospective clinical study was designed to assess the frequency and severity of intravascular hemolysis in patients with new-generation, normally functioning prosthetic heart valves. METHODS: Hemolysis was evaluated in 172 patients with a mechanical prosthesis (53 CarboMedics and 119 Sorin Bicarbon) and in 106 patients with a bioprosthesis (15 St Jude Medical Toronto, 19 Baxter Perimount, and 72 Medtronic Mosaic) in the aortic position, mitral position, or both. Aortic valve replacement was performed in 206 patients, mitral valve replacement in 59 patients, and double valve replacement in 13 patients. The presence of hemolysis was assessed on the basis of the level of serum lactic dehydrogenase and serum haptoglobin and the presence and amount of reticulocytes and schistocytes in the peripheral blood. Severity of intravascular hemolysis was estimated on the basis of serum lactic dehydrogenase. Clinical, echocardiographic, and hematologic evaluations were performed 1, 6, and 12 months after discharge. RESULTS: None of the 278 patients experienced decompensated anemia, whereas at 12 months, mild subclinical hemolysis was identified in 49 patients, 44 (26%) with a mechanical prosthesis and 5 (5%) with a bioprosthesis (P <.001). At multivariate analysis, independent predictors of the presence of subclinical hemolysis were mitral valve replacement (P <.001), use of a mechanical prosthesis (P =.002), and double valve replacement (P =.02). Frequency of hemolysis in patients with stented aortic bioprostheses was 3%, whereas it was absent in those with stentless valves. Among mechanical valve recipients, double versus single valve replacement (P =.04) and mitral versus aortic valve replacement (P =.05) were correlated with the presence of hemolysis; double valve recipients also showed a more severe degree of hemolysis (P =.03). In patients with a Sorin Bicarbon prosthesis, hemolysis was less frequent (22% vs 34%, P =.09) and severe (P <.001) than in those with a CarboMedics prosthesis. CONCLUSIONS: In normally functioning prosthetic heart valves, subclinical hemolysis is a frequent finding. A low incidence of hemolysis is found in stented biologic prostheses, and it is absent in stentless aortic valves. Modifications of valve design may contribute to minimize the occurrence of hemolysis in mechanical prostheses. PMID- 11882832 TI - Myectomy site thrombus formation: an underappreciated source of thromboembolism after septal myectomy. PMID- 11882831 TI - Transitory immunologic response after implantation of the DeBakey VAD continuous axial-flow pump. AB - BACKGROUND: The development of local and systemic infection is a significant risk factor associated with implantation of a ventricular assist device. The immunologic consequence of continuous-flow rotary blood pumps is not known. METHODS: Six male adult patients (mean age 47 plus minus 10.3) with end-stage left heart failure received a DeBakey VAD axial-flow pump for use as a bridge to transplantation. (Four patients underwent transplantation after a mean 115 plus minus 14 days; 2 patients are still waiting for the allograft.) RESULTS: We prospectively monitored T-cell populations and apoptosis-specific aberrant T-cell activation via CD95 triggering and annexin V binding to lymphocytes, identifying T cells undergoing early phases of apoptosis, within the first 10 weeks. Moreover, soluble death-inducing receptors soluble CD95 and soluble tumor necrosis factor-R1 were evaluated by enzyme-linked immunosorbent assay. CONCLUSION: Patients bridged to transplantation by a nonpulsatile ventricular assist device demonstrated an initial pronounced apoptosis-specific immune alteration by increased annexin V binding to CD3 T cells and death-inducing receptors soluble CD95/tumor necrosis factor-R1 (all P <.001). All parameters normalized after 7 weeks to baseline. No blood-borne sepsis was detected, as defined by blood culture, within the first 10 weeks of the cohort study. These results indicate a biphasic immunologic response in patients with end-stage heart failure treated with nonpulsatile ventricular assist devices. PMID- 11882833 TI - Combined surgery for lipomatous hypertrophy of the interatrial septum and pulmonary carcinoma. PMID- 11882834 TI - Fluctuating consciousness caused by hydrocephalus: a complication of aortic valve replacement. PMID- 11882835 TI - Intra-aortic extension of ductal tissue. PMID- 11882836 TI - Type A aortic dissection with associated anomaly of the carotid and vertebral vessels. PMID- 11882837 TI - Paradoxical cerebral emboli of hypernephroma metastatic to the right ventricle five years after primary tumor resection. PMID- 11882838 TI - Homograft aortic root with prosthetic extension as a treatment for aneurysm of the proximal aorta in elderly patients. PMID- 11882839 TI - A surgically treated arteriovenous fistula between the vertebral artery and internal jugular vein after insertion of a central venous catheter for mitral valve replacement. PMID- 11882840 TI - Partial ex situ surgery of the heart. PMID- 11882841 TI - Unifocalization of the neck arteries combined with aortic arch replacement for Williams syndrome. PMID- 11882842 TI - Pulmonary artery sarcoma. PMID- 11882843 TI - Impact of the duration of adjunctive hypothermic circulatory arrest on neurologic outcome with antegrade cerebral perfusion. PMID- 11882844 TI - The first experiments to use resuscitated human cadaver hearts as donor organs. PMID- 11882845 TI - Cellular myoplasty: what are we really trying to achieve? PMID- 11882847 TI - Neurologic hand complications after radial artery harvest for coronary artery bypass grafting. PMID- 11882848 TI - Aortic arch replacement in acute aortic dissection. PMID- 11882849 TI - Supporting good citizenship? PMID- 11882850 TI - How to encourage the right behaviour. PMID- 11882851 TI - Britain banks on embryonic stem cells to gain competitive edge. PMID- 11882852 TI - Call for cloning ban splits UN. PMID- 11882855 TI - Stem-cell reverse angers Australian biologists. PMID- 11882856 TI - Building the triangle: North Carolina. PMID- 11882857 TI - Protests fail to block mountain-lion surveys. PMID- 11882860 TI - Army HIV vaccine to undergo clinical trial as rival is halted. PMID- 11882861 TI - Bubble fusion dispute reaches boiling point. PMID- 11882863 TI - Amazing grace. PMID- 11882864 TI - Music, maestro, please! PMID- 11882865 TI - Curtain has fallen on hopes of legal bioprospecting. PMID- 11882866 TI - Don't fight fire with fire. PMID- 11882871 TI - Nature and function. PMID- 11882873 TI - Applied physics: spin spotting. PMID- 11882872 TI - Biodiversity: biodiversity equals instability? PMID- 11882874 TI - Neurobiology: a cool ion channel. PMID- 11882877 TI - Astronomy: how big stars are made. PMID- 11882876 TI - Biogeochemistry: that's life? PMID- 11882878 TI - Cell biology: ripping up the nuclear envelope. PMID- 11882881 TI - Obituary: Robert Hanbury Brown (1916-2002). PMID- 11882879 TI - Human evolution: tangled genetic routes. PMID- 11882882 TI - Remote sensing: searching for new islands in sea ice. AB - Tobias Island, discovered in 1993 by the German research vessel RV Polarstern, is a system of low-lying banks and shoals hidden in sea ice 70 km off the northeastern coast of Greenland. Here we use satellite radar interferometry and airborne laser scanning to show that this island is 2 km long and 35 m high --- much larger than was originally reported. We have also been able to pinpoint the exact location of a stable area where a new group of small islands may be hidden. This demonstrates that satellite radar interferometry is an effective tool for finding ice-covered islands as well as for mapping them. PMID- 11882883 TI - Palaeontology: 'modern' feathers on a non-avian dinosaur. AB - Discoveries of integumentary coverings on non-avian theropod dinosaurs are becoming commonplace. But the only definitive evidence so far that any of these animals had feathers as we know them today has come from the oviraptorosaur Caudipteryx and the enigmatic coleurosaur Protarchaeopteryx, both of which are considered by some to be secondarily flightless birds. Here we describe the occurrence of pinnate feathers, which clearly feature a rachis and barbs, on a small, non-avian dromaeosaur from northern China. This finding indicates that feathers of modern aspect evolved in dinosaurs before the emergence of birds and flight. PMID- 11882884 TI - Fibre science: supercontraction stress in wet spider dragline. AB - Unrestrained spider dragline 'super-contracts' when it is wetted, causing its length to shrink by about half and its diameter to almost double. Here we measure the supercontraction stresses generated upon initial exposure of spider dragline to moisture and find that they are transient, as well as being greater than previously estimated. Our findings cast doubt on suggestions that supercontraction may help to maintain tension in wet webs and could limit the potential load-bearing applications of silk and its analogues. PMID- 11882885 TI - Anhydrobiosis: plant desiccation gene found in a nematode. AB - When subjected to drought conditions, some organisms enter a state of suspended animation known as anhydrobiosis, surviving for indefinite periods until rehydration allows them to resume normal metabolism. We have identified a gene in the anhydrobiotic nematode Aphelenchus avenae that is upregulated in response to desiccation stress and whose encoded protein shares sequence similarity with a late-embryonic gene that is induced in many plants when they are deprived of water. This finding suggests that animals and plants that undergo anhydrobiosis may use common protective strategies against dehydration, and provides a unifying insight into the mechanism of anhydrobiosis. PMID- 11882886 TI - Determining the composition of the Earth. AB - A long-standing question in the planetary sciences asks what the Earth is made of. For historical reasons, volatile-depleted primitive materials similar to current chondritic meteorites were long considered to provide the 'building blocks' of the terrestrial planets. But material from the Earth, Mars, comets and various meteorites have Mg/Si and Al/Si ratios, oxygen-isotope ratios, osmium isotope ratios and D/H, Ar/H2O and Kr/Xe ratios such that no primitive material similar to the Earth's mantle is currently represented in our meteorite collections. The 'building blocks' of the Earth must instead be composed of unsampled 'Earth chondrite' or 'Earth achondrite'. PMID- 11882887 TI - Out of Africa again and again. AB - The publication of a haplotype tree of human mitochondrial DNA variation in 1987 provoked a controversy about the details of recent human evolution that continues to this day. Now many haplotype trees are available, and new analytical techniques exist for testing hypotheses about recent evolutionary history using haplotype trees. Here I present formal statistical analysis of human haplotype trees for mitochondrial DNA, Y-chromosomal DNA, two X-linked regions and six autosomal regions. A coherent picture of recent human evolution emerges with two major themes. First is the dominant role that Africa has played in shaping the modern human gene pool through at least two--not one--major expansions after the original range extension of Homo erectus out of Africa. Second is the ubiquity of genetic interchange between human populations, both in terms of recurrent gene flow constrained by geographical distance and of major population expansion events resulting in interbreeding, not replacement. PMID- 11882889 TI - Massive star formation in 100,000 years from turbulent and pressurized molecular clouds. AB - Massive stars (with mass m* > 8 solar masses Mmiddle dot in circle) are fundamental to the evolution of galaxies, because they produce heavy elements, inject energy into the interstellar medium, and possibly regulate the star formation rate. The individual star formation time, t*f, determines the accretion rate of the star; the value of the former quantity is currently uncertain by many orders of magnitude, leading to other astrophysical questions. For example, the variation of t*f with stellar mass dictates whether massive stars can form simultaneously with low-mass stars in clusters. Here we show that t*f is determined by the conditions in the star's natal cloud, and is typically about 105yr. The corresponding mass accretion rate depends on the pressure within the cloud--which we relate to the gas surface density--and on both the instantaneous and final stellar masses. Characteristic accretion rates are sufficient to overcome radiation pressure from about 100M middle dot in circle protostars, while simultaneously driving intense bipolar gas outflows. The weak dependence of t*f on the final mass of the star allows high- and low-mass star formation to occur nearly simultaneously in clusters. PMID- 11882888 TI - Identification of a cold receptor reveals a general role for TRP channels in thermosensation. AB - The cellular and molecular mechanisms that enable us to sense cold are not well understood. Insights into this process have come from the use of pharmacological agents, such as menthol, that elicit a cooling sensation. Here we have characterized and cloned a menthol receptor from trigeminal sensory neurons that is also activated by thermal stimuli in the cool to cold range. This cold- and menthol-sensitive receptor, CMR1, is a member of the TRP family of excitatory ion channels, and we propose that it functions as a transducer of cold stimuli in the somatosensory system. These findings, together with our previous identification of the heat-sensitive channels VR1 and VRL-1, demonstrate that TRP channels detect temperatures over a wide range and are the principal sensors of thermal stimuli in the mammalian peripheral nervous system. PMID- 11882890 TI - Coherent emission of light by thermal sources. AB - A thermal light-emitting source, such as a black body or the incandescent filament of a light bulb, is often presented as a typical example of an incoherent source and is in marked contrast to a laser. Whereas a laser is highly monochromatic and very directional, a thermal source has a broad spectrum and is usually quasi-isotropic. However, as is the case with many systems, different behaviour can be expected on a microscopic scale. It has been shown recently that the field emitted by a thermal source made of a polar material is enhanced by more than four orders of magnitude and is partially coherent at a distance of the order of 10 to 100nm. Here we demonstrate that by introducing a periodic microstructure into such a polar material (SiC) a thermal infrared source can be fabricated that is coherent over large distances (many wavelengths) and radiates in well defined directions. Narrow angular emission lobes similar to antenna lobes are observed and the emission spectra of the source depends on the observation angle--the so-called Wolf effect. The origin of the coherent emission lies in the diffraction of surface-phonon polaritons by the grating. PMID- 11882891 TI - A general process for in situ formation of functional surface layers on ceramics. AB - Ceramics are often prepared with surface layers of different composition from the bulk, in order to impart a specific functionality to the surface or to act as a protective layer for the bulk material. Here we describe a general process by which functional surface layers with a nanometre-scale compositional gradient can be readily formed during the production of bulk ceramic components. The basis of our approach is to incorporate selected low-molecular-mass additives into either the precursor polymer from which the ceramic forms, or the binder polymer used to prepare bulk components from ceramic powders. Thermal treatment of the resulting bodies leads to controlled phase separation ('bleed out') of the additives, analogous to the normally undesirable outward loss of low-molecular-mass components from some plastics; subsequent calcination stabilizes the compositionally changed surface region, generating a functional surface layer. This approach is applicable to a wide range of materials and morphologies, and should find use in catalysts, composites and environmental barrier coatings. PMID- 11882892 TI - Observation and interpretation of a time-delayed mechanism in the hydrogen exchange reaction. AB - Extensive theoretical and experimental studies have shown the hydrogen exchange reaction H+H2 --> H2+H to occur predominantly through a 'direct recoil' mechanism: the H--H bonds break and form concertedly while the system passes straight over a collinear transition state, with recoil from the collision causing the H2 product molecules to scatter backward. Theoretical predictions agree well with experimental observations of this scattering process. Indirect exchange mechanisms involving H3 intermediates have been suggested to occur as well, but these are difficult to test because bimolecular reactions cannot be studied by the femtosecond spectroscopies used to monitor unimolecular reactions. Moreover, full quantum simulations of the time evolution of bimolecular reactions have not been performed. For the isotopic variant of the hydrogen exchange reaction, H+D2 --> HD+D, forward scattering features observed in the product angular distribution have been attributed to possible scattering resonances associated with a quasibound collision complex. Here we extend these measurements to a wide range of collision energies and interpret the results using a full time dependent quantum simulation of the reaction, thus showing that two different reaction mechanisms modulate the measured product angular distribution features. One of the mechanisms is direct and leads to backward scattering, the other is indirect and leads to forward scattering after a delay of about 25 femtoseconds. PMID- 11882893 TI - Glacial--interglacial stability of ocean pH inferred from foraminifer dissolution rates. AB - The pH of the ocean is controlled by the chemistry of calcium carbonate. This system in turn plays a large role in regulating the CO2 concentration of the atmosphere on timescales of thousands of years and longer. Reconstructions of ocean pH and carbonate-ion concentration are therefore needed to understand the ocean's role in the global carbon cycle. During the Last Glacial Maximum (LGM), the pH of the whole ocean is thought to have been significantly more basic, as inferred from the isotopic composition of boron incorporated into calcium carbonate shells, which would partially explain the lower atmospheric CO2 concentration at that time. Here we reconstruct carbonate-ion concentration--and hence pH--of the glacial oceans, using the extent of calcium carbonate dissolution observed in foraminifer faunal assemblages as compiled in the extensive global CLIMAP data set. We observe decreased carbonate-ion concentrations in the glacial Atlantic Ocean, by roughly 20 micromolkg-1, while little change occurred in the Indian and Pacific oceans relative to today. In the Pacific Ocean, a small (5 micromolkg-1) increase occurred below 3,000m. This rearrangement of ocean pH may be due to changing ocean circulation from glacial to present times, but overall we see no evidence for a shift in the whole-ocean pH as previously inferred from boron isotopes. PMID- 11882894 TI - Laser--Raman imagery of Earth's earliest fossils. AB - Unlike the familiar Phanerozoic history of life, evolution during the earlier and much longer Precambrian segment of geological time centred on prokaryotic microbes. Because such microorganisms are minute, are preserved incompletely in geological materials, and have simple morphologies that can be mimicked by nonbiological mineral microstructures, discriminating between true microbial fossils and microscopic pseudofossil 'lookalikes' can be difficult. Thus, valid identification of fossil microbes, which is essential to understanding the prokaryote-dominated, Precambrian 85% of life's history, can require more than traditional palaeontology that is focused on morphology. By combining optically discernible morphology with analyses of chemical composition, laser--Raman spectroscopic imagery of individual microscopic fossils provides a means by which to address this need. Here we apply this technique to exceptionally ancient fossil microbe-like objects, including the oldest such specimens reported from the geological record, and show that the results obtained substantiate the biological origin of the earliest cellular fossils known. PMID- 11882895 TI - Questioning the evidence for Earth's oldest fossils. AB - Structures resembling remarkably preserved bacterial and cyanobacterial microfossils from about 3,465-million-year-old Apex cherts of the Warrawoona Group in Western Australia currently provide the oldest morphological evidence for life on Earth and have been taken to support an early beginning for oxygen producing photosynthesis. Eleven species of filamentous prokaryote, distinguished by shape and geometry, have been put forward as meeting the criteria required of authentic Archaean microfossils, and contrast with other microfossils dismissed as either unreliable or unreproducible. These structures are nearly a billion years older than putative cyanobacterial biomarkers, genomic arguments for cyanobacteria, an oxygenic atmosphere and any comparably diverse suite of microfossils. Here we report new research on the type and re-collected material, involving mapping, optical and electron microscopy, digital image analysis, micro Raman spectroscopy and other geochemical techniques. We reinterpret the purported microfossil-like structure as secondary artefacts formed from amorphous graphite within multiple generations of metalliferous hydrothermal vein chert and volcanic glass. Although there is no support for primary biological morphology, a Fischer- Tropsch-type synthesis of carbon compounds and carbon isotopic fractionation is inferred for one of the oldest known hydrothermal systems on Earth. PMID- 11882896 TI - Arctic microorganisms respond more to elevated UV-B radiation than CO2. AB - Surface ultraviolet-B radiation and atmospheric CO2 concentrations have increased as a result of ozone depletion and burning of fossil fuels. The effects are likely to be most apparent in polar regions where ozone holes have developed and ecosystems are particularly sensitive to disturbance. Polar plant communities are dependent on nutrient cycling by soil microorganisms, which represent a significant and highly labile portion of soil carbon (C) and nitrogen (N). It was thought that the soil microbial biomass was unlikely to be affected by exposure of their associated plant communities to increased UV-B. In contrast, increasing atmospheric CO2 concentrations were thought to have a strong effect as a result of greater below-ground C allocation. In addition, there is a growing belief that ozone depletion is of only minor environmental concern because the impacts of UV B radiation on plant communities are often very subtle. Here we show that 5 years of exposure of a subarctic heath to enhanced UV-B radiation both alone and in combination with elevated CO2 resulted in significant changes in the C:N ratio and in the bacterial community structure of the soil microbial biomass. PMID- 11882897 TI - Diversity-dependent production can decrease the stability of ecosystem functioning. AB - There is concern that species loss may adversely affect ecosystem functioning and stability. But although there is evidence that biodiversity loss can lead to reductions in biomass production, there is no direct evidence that biodiversity loss affects ecosystem resistance (ability to withstand perturbation) or resilience (recovery from perturbation). Yet theory, laboratory experiments and indirect experimental evidence strongly suggest that diversity and stability are related. Here we report results from a field experiment with factorially crossed perturbation and diversity manipulations. We simulated drought perturbation on constructed grassland ecosystems containing 1, 2, 4, 8 or 32 plant species. Under unperturbed conditions, the species-poor systems achieved lower biomass production than the species-rich systems. However, the species-poor systems were more resistant to perturbation than the species-rich systems. The species-poor systems also showed a larger initial resilience following perturbation, although the original relationship between diversity and productivity was fully restored after 1year. Our results confirm that biodiversity increases biomass production, but they also point to the fact that such diversity--production associations may lead to an inverse relationship between biodiversity and the stability of ecosystem functioning. PMID- 11882898 TI - Chimaeric sounds reveal dichotomies in auditory perception. AB - By Fourier's theorem, signals can be decomposed into a sum of sinusoids of different frequencies. This is especially relevant for hearing, because the inner ear performs a form of mechanical Fourier transform by mapping frequencies along the length of the cochlear partition. An alternative signal decomposition, originated by Hilbert, is to factor a signal into the product of a slowly varying envelope and a rapidly varying fine time structure. Neurons in the auditory brainstem sensitive to these features have been found in mammalian physiological studies. To investigate the relative perceptual importance of envelope and fine structure, we synthesized stimuli that we call 'auditory chimaeras', which have the envelope of one sound and the fine structure of another. Here we show that the envelope is most important for speech reception, and the fine structure is most important for pitch perception and sound localization. When the two features are in conflict, the sound of speech is heard at a location determined by the fine structure, but the words are identified according to the envelope. This finding reveals a possible acoustic basis for the hypothesized 'what' and 'where' pathways in the auditory cortex. PMID- 11882899 TI - Long-term plasticity in hippocampal place-cell representation of environmental geometry. AB - The hippocampus is widely believed to be involved in the storage or consolidation of long-term memories. Several reports have shown short-term changes in single hippocampal unit activity during memory and plasticity experiments, but there has been no experimental demonstration of long-term persistent changes in neuronal activity in any region except primary cortical areas. Here we report that, in rats repeatedly exposed to two differently shaped environments, the hippocampal place-cell representations of those environments gradually and incrementally diverge; this divergence is specific to environmental shape, occurs independently of explicit reward, persists for periods of at least one month, and transfers to new enclosures of the same shape. These results indicate that place cells may be a neural substrate for long-term incidental learning, and demonstrate the long term stability of an experience-dependent firing pattern in the hippocampal formation. PMID- 11882900 TI - Balanced responsiveness to chemoattractants from adjacent zones determines B-cell position. AB - B lymphocytes re-circulate between B-cell-rich compartments (follicles or B zones) in secondary lymphoid organs, surveying for antigen. After antigen binding, B cells move to the boundary of B and T zones to interact with T-helper cells. Despite the importance of B--T-cell interactions for the induction of antibody responses, the mechanism causing B-cell movement to the T zone has not been defined. Here we show that antigen-engaged B cells have increased expression of CCR7, the receptor for the T-zone chemokines CCL19 and CCL21, and that they exhibit increased responsiveness to both chemoattractants. In mice lacking lymphoid CCL19 and CCL21 chemokines, or with B cells that lack CCR7, antigen engagement fails to cause movement to the T zone. Using retroviral-mediated gene transfer we demonstrate that increased expression of CCR7 is sufficient to direct B cells to the T zone. Reciprocally, overexpression of CXCR5, the receptor for the B-zone chemokine CXCL13, is sufficient to overcome antigen-induced B-cell movement to the T zone. These findings define the mechanism of B-cell relocalization in response to antigen, and establish that cell position in vivo can be determined by the balance of responsiveness to chemoattractants made in separate but adjacent zones.